PMID- 17107561 TI - Whole genome analysis of the marine Bacteroidetes'Gramella forsetii' reveals adaptations to degradation of polymeric organic matter. AB - Members of the Bacteroidetes, formerly known as the Cytophaga-Flavobacteria Bacteroides (CFB) phylum, are among the major taxa of marine heterotrophic bacterioplankton frequently found on macroscopic organic matter particles (marine snow). In addition, they have been shown to also represent a significant part of free-living microbial assemblages in nutrient-rich microenvironments. Their abundance and distribution pattern in combination with enzymatic activity studies has led to the notion that organisms of this group are specialists for degradation of high molecular weight compounds in both the dissolved and particulate fraction of the marine organic matter pool, implying a major role of Bacteroidetes in the marine carbon cycle. Despite their ecological importance, comprehensive molecular data on organisms of this group have been scarce so far. Here we report on the first whole genome analysis of a marine Bacteroidetes representative, 'Gramella forsetii' KT0803. Functional analysis of the predicted proteome disclosed several traits which in joint consideration suggest a clear adaptation of this marine Bacteroidetes representative to the degradation of high molecular weight organic matter, such as a substantial suite of genes encoding hydrolytic enzymes, a predicted preference for polymeric carbon sources and a distinct capability for surface adhesion. PMID- 17107560 TI - Phylogenetic identification and metabolism of marine dimethylsulfide-consuming bacteria. AB - Microbial consumption is one of the main processes, along with photolysis and ventilation, that remove the biogenic trace gas dimethylsulfide (DMS) from the surface ocean. Although a few isolates of marine bacteria have been studied for their ability to utilize DMS, little is known about the characteristics or phylogenetic affiliation of DMS consumers in seawater. We enriched coastal and open-ocean waters with different carbon sources to stimulate different bacterial communities (glucose-consuming bacteria, methyl group-consuming bacteria and DMS consumers) in order to test how this affected DMS consumption and to examine which organisms might be involved. Dimethylsulfide consumption was greatly stimulated in the DMS addition treatments whereas there was no stimulation in the other treatments. Analysis of microbial DNA by two different techniques (sequenced bands from DGGE gels and clone libraries) showed that bacteria grown specifically with the presence of DMS were closely related to the genus Methylophaga. We also followed the fate of consumed DMS in some of the enrichments. Dimethylsulfide was converted mostly to DMSO in glucose or methanol enrichments, whereas it was converted mostly to sulfate in DMS enrichments, the latter suggesting use of DMS as a carbon and energy source. Our results indicate that unlike the biochemical precursor of DMS, dimethylsulfoniopropionate (DMSP), which is consumed by a broad spectrum of marine microorganisms, DMS seems to be utilized as a carbon and electron source by specialists. This is consistent with the usual observation that DMSP turns over at much higher rates than DMS. PMID- 17107562 TI - Fungal hydroquinones contribute to brown rot of wood. AB - The fungi that cause brown rot of wood initiate lignocellulose breakdown with an extracellular Fenton system in which Fe(2+) and H(2)O(2) react to produce hydroxyl radicals (.OH), which then oxidize and cleave the wood holocellulose. One such fungus, Gloeophyllum trabeum, drives Fenton chemistry on defined media by reducing Fe(3+) and O(2) with two extracellular hydroquinones, 2,5 dimethoxyhydroquinone (2,5-DMHQ) and 4,5-dimethoxycatechol (4,5-DMC). However, it has never been shown that the hydroquinones contribute to brown rot of wood. We grew G. trabeum on spruce blocks and found that 2,5-DMHQ and 4,5-DMC were each present in the aqueous phase at concentrations near 20 microM after 1 week. We determined rate constants for the reactions of 2,5-DMHQ and 4,5-DMC with the Fe(3+)-oxalate complexes that predominate in wood undergoing brown rot, finding them to be 43 l mol(-1) s(-1) and 65 l mol(-1) s(-1) respectively. Using these values, we estimated that the average amount of hydroquinone-driven .OH production during the first week of decay was 11.5 micromol g(-1) dry weight of wood. Viscometry of the degraded wood holocellulose coupled with computer modelling showed that a number of the same general magnitude, 41.2 micromol oxidations per gram, was required to account for the depolymerization that occurred in the first week. Moreover, the decrease in holocellulose viscosity was correlated with the measured concentrations of hydroquinones. Therefore, hydroquinone-driven Fenton chemistry is one component of the biodegradative arsenal that G. trabeum expresses on wood. PMID- 17107563 TI - Diversity and expression of nitrogenase genes (nifH) from ectomycorrhizas of Corsican pine (Pinus nigra). AB - The diversity of bacterial nitrogenase genes (nifH) and their mRNA transcription in ectomycorrhizas of Corsican pine (Pinus nigra) were examined. DNA and RNA were extracted from surface-sterilized and non-sterilized Corsican pine roots colonized by the ectomycorrhizal (ECM) fungi, Suillus variegatus and Tomentellopsis submollis. DNA-derived nifH polymerase chain reaction (PCR) products were obtained from all samples, but only a few reverse transcription PCRs for nifH mRNA were successful, suggesting that nitrogenase genes were not always transcribed. Several different nifH sequences were detected and the bacteria actively transcribing nifH were different from those whose genes were detected through DNA-based PCR. Putative nitrogenase amino acid sequences revealed that more than half of the nifH products were derived from methylotrophic bacteria, such as Methylocella spp. The next most frequent sequence types were similar to those from Burkholderia. PMID- 17107565 TI - Redefining the education of the advanced practice psychiatric nurse. PMID- 17107566 TI - Advanced practice psychiatric mental health nursing, finding our core: the therapeutic relationship in 21st century. AB - TOPIC: Increasingly, students from various professional backgrounds are enrolling in Psychiatric Mental Health (PMH) Nursing graduate programs, especially at the post-master's level. Faculty must educate these students to provide increasingly complex care while socializing them as PMH advanced practitioners. PURPOSE: To present how one online program is addressing these issues by reasserting the centrality of the relationship and by assuring it has at least equal footing with the application of a burgeoning knowledge base of neurobiology of mental illness. SOURCES: Published literature from nursing and psychology. CONCLUSIONS: The PMH graduate faculty believes that they have developed strategies to meet this challenge and to help build a PMH workforce that will maintain the centrality of the relationship in PMH practice. PMID- 17107567 TI - A framework for cultural competence in advanced practice psychiatric and mental health education. AB - TOPIC: Cultural competence in psychiatric advanced practice education. PURPOSE: To present a framework for educators to use when addressing culturally competent advanced psychiatric nursing practice. SOURCES: NONPF competencies, published literature, and Web resources. CONCLUSIONS: Development of cultural competence is an important link to the reduction of mental health disparities. Situating the advanced nursing practice process within a negotiating space allows for incorporation of cultural information into patient care. The framework presented in this paper provides a theoretical and practical approach to orienting students to meeting the elements of cultural competence set out in the NONPF recommendations. PMID- 17107568 TI - Peer consultation: a unique experiential model for P/MHNP students. AB - TOPIC: Peer consultation: A unique experiential model for P/MHNP students. PURPOSE: The purpose of this paper is to describe the consultation component of the Adult Psychiatric Mental Health Graduate Program at UMASS Lowell. The consultation requirement is described in the Professional Role competency as "provides consultation to healthcare providers and others to enhance quality and cost-effective services for patients and to effect change in organizational systems" (NONPF, 2003, p. 10). The activity described in this paper also includes demonstration of components of the competencies for assessment, diagnosis, plan of care, and teaching/coaching (NONPF, 2003). SOURCES: This paper is based on review of the literature and the author's clinical and teaching experience. CONCLUSIONS: Form of classroom instruction is a unique way of assisting students in the development of the professional role as a consultant. PMID- 17107569 TI - An introductory clinical core course in psychiatric management: an innovative lifespan course blending all nurse practitioner majors. AB - TOPIC: The prevalence of anxiety, depression, substance abuse, and suicidal ideation is significant in primary care settings across the country. Nonpsychiatric nurse practitioners must be able to recognize symptoms of common psychiatric disorders, know how to treat less complex mental illnesses, and know when to refer to psychiatric mental health nurse practitioners (PMHNPs). PURPOSE: This article describes the course content, assignments, and teaching strategies used in a clinical core course in the nurse practitioner (NP) curriculum that is required for all NP majors at the University of Texas at Arlington. Psychiatric Management for Advanced Practice provides the foundation for later PMHNP major specific clinical courses. SOURCES: Development of the course content was based on NONPF Domains and Competencies for the NP, input from graduate NP faculty using a modified Delphi approach, NP student feedback, review of curriculum from other schools, and review of the literature on depression, suicide, anxiety, and substance abuse disorders in primary care settings. CONCLUSIONS: Since 1999, students from the eight different NP programs at the University of Texas at Arlington have been required to take this course. Student, faculty, and graduate feedback about this course have been consistently positive. Many NP students comment on feeling much more comfortable assessing for depression, suicide, and substance use. PMID- 17107570 TI - What does it take to make a nurse? considerations of the CNL and DNP role development. PMID- 17107571 TI - Smoke, smoke, smoke that cigarette. PMID- 17107572 TI - Creating a healing environment. PMID- 17107573 TI - The devil and Daniel Johnston, Jeff Feuerzeig (director). PMID- 17107586 TI - Abstracts from the Asian Pacific Digestion Week 2006. November 26-29, 2006. Lahug Cebu City, Philippines. PMID- 17107593 TI - Adenosine A2A receptors are up-regulated in Pick's disease frontal cortex. AB - Adenosine A2A receptors (A2AR) are highly expressed in striatum. However, they are also present in extrastriatal structures. A2AR were studied in post-mortem human frontal cortex from Pick's disease (PiD) and age-matched non-demented controls by radioligand binding assays, Western-blotting, real-time PCR and adenylyl cyclase activity determination. Saturation binding assay using [3H]ZM 241385, a selective A2A antagonist, as radioligand revealed a significant increase in total adenosine A2AR numbers (Bmax) in frontal cortex from PiD samples (191% of control Bmax), suggesting up-regulation of this receptor. A significant increase in the level of A2AR was also detected by Western-blotting. Furthermore, expression of mRNA coding A2AR determined by quantitative real-time PCR was enhanced. In agreement, stimulation of adenylyl cyclase by CGS 21680, a selective A2A receptor agonist, was significantly strengthened. Up-regulation of A2B receptors and their corresponding mRNA was also observed. These results show that A2A adenosine receptor/adenylyl cyclase transduction pathway is up-regulated and sensitized in frontal cortex brain from PiD. PMID- 17107595 TI - Myofiber degeneration in autosomal dominant Emery-Dreifuss muscular dystrophy (AD EDMD) (LGMD1B). AB - Autosomal dominant Emery-Dreifuss muscular dystrophy is caused by mutations in the LMNA gene that code for the nuclear membrane protein lamin A/C. We investigated skeletal muscle fibers from several muscles for cytoplasmic degenerative changes in three patients with genetically confirmed Emery-Dreifuss muscular dystrophy. Methods included quantitative light and electron microscopy and PCR-based mutational analysis. RESULTS: The degenerative pathway was characterized by the gradual replacement of individual myofibers by connective tissue. Early stages of degeneration typically involved only a segment of the cross-sectional area of a myofiber. Intermediate stages consisted of myofiber shrinkage due to "shedding" of peripheral cytoplasmic portions into the endomysial space, and fragmentation of the myofibers by interposed collagen fibrils. Empty basement membrane sheaths surrounded by abundant deposits of extracellular matrix marked the end stage of the degenerative process. The nuclear number-to-cytoplasmic area in myofibers of one patient increased with increasing cross-sectional area, suggesting that satellite cell fusion with myofibers may have compensated for myofiber shrinkage. The pattern of degeneration described herein differs from muscular dystrophies with plasma membrane defects (dystrophinopathy, dysferlinopathy) and explains the frequently found absence of highly elevated serum creatine kinase levels in autosomal dominant Emery-Dreifuss muscular dystrophy. PMID- 17107594 TI - CD45 isoform expression in microglia and inflammatory cells in HIV-1 encephalitis. AB - CD45 is a membrane tyrosine phosphatase that modulates the function of the hematopoietic cells. In vitro, agonist antibodies to CD45RO or CD45RB isoforms have been shown to suppress microglial activation, but whether microglia in vivo express these isoforms in HIV encephalitis (HIVE) is unknown. Brain sections from control and HIVE were immunostained for CD45 isoforms using exon-specific antibodies (RA, RB, RC and RO). RA and RC were limited to rare lymphocytes, while RB expression was robust in microglia and inflammatory cells. RO was low in control microglia, but increased in HIVE. RO was also localized to macrophages and CD8+ T cells. Targeting CD45 in vivo with isoform-specific antibodies remains a therapeutic option for neuroinflammatory diseases. PMID- 17107596 TI - Characterization of glioblastomas in young adults. AB - Most adult glioblastoma multiformes (GBMs) present in patients 45-70 years old; tumors occurring at the extremes of the adult age spectrum are uncommon, and seldom studied. We hypothesized that young-adult GBMs would differ from elderly adult and from pediatric GBMs. Cases were identified from years 1997 to 2005. Demographic and histological features, MIB-1 and TP53 immunohistochemical findings and epidermal growth factor receptor (EGFR) amplification status by fluorescence in situ hybridization were compiled and correlated with survival. Twenty-eight (74%) of our 38 young-adult GBM patients had primary de novo tumors, two of which occurred in patients with cancer syndromes. Two additional GBMs were radiation-induced and eight (21%) were secondary GBMs. Seven patients were identified as long-term (>3 years) survivors. Six of 38 cases manifested unusual morphological features, including three epithelioid GBMs, one rhabdoid GBM, one gliosarcoma and one small cell GBM containing abundant, refractile, eosinophilic inclusions. MIB-1 index emerged as the most important prognosticator of survival (P < 0.005). Although there was a trend between extent of necrosis, TP53 immunohistochemical expression, and EGFR amplification status and survival, none reached statistical significance. GBMs in young adults are a more inhomogeneous tumor group than GBMs occurring in older adult patients and show features that overlap with both pediatric and adult GBMs. PMID- 17107598 TI - The quest for remyelination: a new role for neurotrophins and their receptors. AB - The formation of myelin is dependent on a reciprocal and intimate relationship between neurons and the myelin-forming glia. Recently, the neurotrophin family of growth factors has been shown to regulate the complex cell-cell interactions that control myelination. Neurotrophins and their receptors influence myelin formation via two distinct mechanisms, either by acting on the neurons, changing the axonal signals that control myelination, or by acting directly on the myelin-forming glia. In this review, we will discuss research highlighting the ability of neurotrophins to both promote and inhibit the myelination process. As reflected in the work presented here, these effects are dependent on a delicate balance of which neurotrophins are expressed, and what receptors are activated. Additionally, we examine an emerging model in which the growth factors that promote the early survival and differentiation of particular sets of neurons later play important roles as key regulators in glial development. Characterizing the temporal expression and the cellular targets of neurotrophins, both during development and after injury, represents a pivotal step in developing a greater understanding of the myelination process, contributing to the development of effective treatments for demyelinating conditions. We conclude this review by discussing the potential for neurotrophins as therapeutics in the quest for remyelination. PMID- 17107599 TI - Neurotrophic factors in neurodegeneration. AB - Neurotrophic factors (NTFs) have the unique potential to support neuronal survival and to augment neuronal function in the injured and diseased nervous system. Numerous studies conducted over the last 20 years have provided evidence for the potent therapeutic potential of NTFs in animal models of neurodegenerative diseases. However, major obstacles for the therapeutic use of NTFs are the inability to deliver proteins across the blood-brain-barrier, and dose-limiting adverse effects resulting from the broad exposure of nontargeted structures to NTFs. Two recent developments have allowed NTFs' promise to be truly tested for the first time: first, recent improvements in viral vectors that allow the targeted delivery of NTFs while providing a long-lasting supply and sufficient therapeutic doses of NTFs; and second, improved animal models developed in recent years. In this review, we will discuss some of the potential therapeutic applications of NTFs in neurodegenerative diseases and the potential contribution of disturbed neurotrophic factor signaling to neurodegenerative diseases. PMID- 17107600 TI - The dark side of the NGF family: neurotrophins in neoplasias. AB - Although neurotrophins of the nerve growth factor (NGF) family are best known for their neurite outgrowth-inducing and survival-promoting effects on neuronal cells, these are actually pleiotropic growth factors acting physiologically on many different cell types of our body. As for many other growth factors, dysregulation of neurotrophin signal transduction is found in a number of tumors where they can accompany or contribute to malignant transformation. Interestingly, tropomyosin-related kinase (Trk) receptor activation can either support or suppress tumor growth, depending on the tumor type. These same divergent responses have been observed with neurotrophins binding to the p75NTR neurotrophin receptor on tumor cells. This article summarizes the current knowledge on the role of neurotrophins and their receptors in malignancies, with special focus on tumors of neuropathological interest. PMID- 17107601 TI - Neurotrophins and peripheral neuropathies. AB - Peripheral neuropathy is a common disorder seen in general neurology and neuromuscular specialty clinics. Treatment options directed at the underlying cause can only be offered in a handful of conditions, such as those with possible autoimmune etiology. The remainder fall into the idiopathic or genetic category with no known treatment. This review surveys the evidence supporting the rationale for the therapeutic use of neurotrophins and other neurotrophic factors in these disorders in relationship to the underlying pathobiological process. Previous clinical trials are assessed, and increasingly better understood and appreciated therapeutic potential of neurotrophins is emphasized. PMID- 17107602 TI - Pathophysiological mechanisms for actions of the neurotrophins. AB - Neurotrophins provide trophic and tropic support for different neuronal subpopulations in the developing and adult nervous systems. Expression of the neurotrophins and their receptors can be altered in several different disease or injury states that impact upon the functions in the central and peripheral nervous systems. The intracellular signals used by the neurotrophins are triggered by ligand binding to the cell surface Trk and p75NTR receptors. In general, signals emanating from Trk receptors support survival, growth and synaptic strengthening, while those emanating from p75NTR induce apoptosis, attenuate growth and weaken synaptic signaling. Mature neurotrophins are the preferred ligand for Trk proteins while p75NTR binds preferentially to the proneurotrophins and serves as a signaling component of the receptor complex for growth inhibitory molecules of central nervous system myelin [ie, myelin associated glycoprotein (MAG), oligodendrocyte-myelin glycoprotein (OMgP) and Nogo]. The functional antagonism between Trk and p75NTR signaling may significantly impact the pathogenesis of human neurodevelopmental and neurodegenerative diseases and further complicate therapeutic uses of exogenous neurotrophins. The potential for each is discussed in this review. PMID- 17107603 TI - A 4-year-old girl with mass in the frontotemporal bone. PMID- 17107604 TI - A 57-year-old female with a benign tumor in the ethmoid sinus. PMID- 17107605 TI - An 80-year-old woman with neuropsychiatric changes and clinical symptoms of encephalitis. PMID- 17107607 TI - Physicochemical attack against solid tumors based on the reversal of direction of entropy flow: an attempt to introduce thermodynamics in anticancer therapy. AB - BACKGROUND: There are many differences between healthy tissue and growing tumor tissue, including metabolic, structural and thermodynamic differences. Both structural and thermodynamic differences can be used to follow the entropy differences in cancerous and normal tissue. Entropy production is a bilinear form of the rates of irreversible processes and the corresponding "generalized forces". Entropy production due to various dissipation mechanisms based on temperature differences, chemical potential gradient, chemical affinity, viscous stress and exerted force is a promising tool for calculations relating to potential targets for tumor isolation and demarcation. METHODS: The relative importance of five forms of entropy production was assessed through mathematical estimation. Using our mathematical model we demonstrated that the rate of entropy production by a cancerous cell is always higher than that of a healthy cell apart from the case of the application of external energy. Different rates of entropy production by two kinds of cells influence the direction of entropy flow between the cells. Entropy flow from a cancerous cell to a healthy cell transfers information regarding the cancerous cell and propagates its invasive action to the healthy tissues. To change the direction of entropy flow, in addition to designing certain biochemical pathways to reduce the rate of entropy production by cancerous cells, we suggest supplying external energy to the tumor area, changing the relative rate of entropy production by the two kinds of cells and leading to a higher entropy accumulation in the surrounding normal cells than in the tumorous cells. CONCLUSION: Through the use of mathematical models it was quantitatively demonstrated that when no external force field is applied, the rate of entropy production of cancerous cells is always higher than that of healthy cells. However, when the external energy of square wave electric pulses is applied to tissues, the rate of entropy production of normal cells may exceed that of cancerous cells. Consequently, the application of external energy to the body can reverse the direction of the entropy current. The harmful effect brought about by the entropy flow from cancerous to healthy tissue can be blocked by the reversed direction of entropy current from the irradiated normal tissue around the tumor. PMID- 17107606 TI - Metabolic network driven analysis of genome-wide transcription data from Aspergillus nidulans. AB - BACKGROUND: Aspergillus nidulans (the asexual form of Emericella nidulans) is a model organism for aspergilli, which are an important group of filamentous fungi that encompasses human and plant pathogens as well as industrial cell factories. Aspergilli have a highly diversified metabolism and, because of their medical, agricultural and biotechnological importance, it would be valuable to have an understanding of how their metabolism is regulated. We therefore conducted a genome-wide transcription analysis of A. nidulans grown on three different carbon sources (glucose, glycerol, and ethanol) with the objective of identifying global regulatory structures. Furthermore, we reconstructed the complete metabolic network of this organism, which resulted in linking 666 genes to metabolic functions, as well as assigning metabolic roles to 472 genes that were previously uncharacterized. RESULTS: Through combination of the reconstructed metabolic network and the transcription data, we identified subnetwork structures that pointed to coordinated regulation of genes that are involved in many different parts of the metabolism. Thus, for a shift from glucose to ethanol, we identified coordinated regulation of the complete pathway for oxidation of ethanol, as well as upregulation of gluconeogenesis and downregulation of glycolysis and the pentose phosphate pathway. Furthermore, on change in carbon source from glucose to ethanol, the cells shift from using the pentose phosphate pathway as the major source of NADPH (nicotinamide adenine dinucleotide phosphatase, reduced form) for biosynthesis to use of the malic enzyme. CONCLUSION: Our analysis indicates that some of the genes are regulated by common transcription factors, making it possible to establish new putative links between known transcription factors and genes through clustering. PMID- 17107608 TI - Serial detection of circulating tumour cells by reverse transcriptase-polymerase chain reaction assays is a marker for poor outcome in patients with malignant melanoma. AB - BACKGROUND: Detection of circulating malignant cells (CMCs) through a reverse transcriptase-polymerase chain reaction (RT-PCR) assay seems to be a demonstration of systemic disease. We here evaluated the prognostic role of RT PCR assays in serially-taken peripheral blood samples from patients with malignant melanoma (MM). METHODS: One hundred forty-nine melanoma patients with disease stage ranging from I to III were consecutively collected in 1997. A multi marker RT-PCR assay was used on peripheral blood samples obtained at time of diagnosis and every 6 months during the first two years of follow-up (total: 5 samples). Univariate and multivariate analyses were performed after 83 months of median follow-up. RESULTS: Detection of at least one circulating mRNA marker was considered a signal of the presence of CMC (referred to as PCR-positive assay). A significant correlation was found between the rate of recurrences and the increasing number of PCR-positive assays (P = 0.007). Presence of CMC in a high number (> or =2) of analysed blood samples was significantly correlated with a poor clinical outcome (disease-free survival: P = 0.019; overall survival: P = 0.034). Multivariate analysis revealed that presence of a PCR-positive status does play a role as independent prognostic factors for overall survival in melanoma patients, adding precision to the predictive power of the disease stage. CONCLUSION: Our findings indicated that serial RT-PCR assay may identify a high risk subset of melanoma patients with occult cancer cells constantly detected in blood circulation. Prolonged presence of CMCs seems to act as a surrogate marker of disease progression or a sign of more aggressive disease. PMID- 17107609 TI - Treatment of polydrug-using opiate dependents during withdrawal: towards a standardisation of treatment. AB - BACKGROUND: The growing tendency among opioid addicts to misuse multiple other drugs should lead clinicians and researchers to search for new pharmacological strategies in order to prevent life-threatening complications and minimize withdrawal symptoms during polydrug detoxification. METHODS: A non-randomised, open-label in-patient detoxification study was used to compare the short-time efficacy of a standardised regimen comprising 6 days Buprenorphine and 10 days Valproate (BPN/VPA) (n = 12) to a control group (n = 50) who took a 10-day traditional Clonidine/Carbamazepine (CLN/CBZ) regimen. Sixty-two dependent subjects admitted to a detoxification unit were included, all dependent on at least opioids and benzodiazepines. Other dependencies were not excluded. RESULTS: In the BPN/VPA group, 8 out of 12 patients (67%) completed treatment compared with 25 of 50 patients (50%) in the CLN/CBZ group; this difference between the groups was non-significant (p = 0.15). Withdrawal symptoms were reduced in both groups, but only the BPN/VPA group achieved a reduction in withdrawal symptoms from day one. The difference between the two groups was significantly in favour of the BPN/VPA group for days 2 (p < 0.001), 3 (p < 0.05), 4 (p < 0.001), 5 (p < 0.01), 7 (p < 0.01) and 8 (p < 0.05). The BPN/VPA combination did not affect blood pressure, pulse or liver function, and the total burden of side-effects was experienced as modest. There appeared to be no pharmacological interactions of clinical concern, based on measurement of Buprenorphine and Valproate serum levels. Both the patients and the staff were satisfied with the standardised treatment combination. CONCLUSION: Overall, the combination of Buprenorphine and Valproate seems to be a safe and promising method for treating multiple drug withdrawal symptoms. The results of this study suggest that the BPN/VPA combination is potentially a better detoxification treatment for polydrug withdrawal than the traditional treatment with Clonidine and Carbamazepine. However, a randomised, double-blind study with a larger sample size to confirm our results is recommended. PMID- 17107610 TI - "In vitro" activities of antimycobacterial agents against Mycobacterium avium subsp. paratuberculosis linked to Crohn's disease and paratuberculosis. PMID- 17107611 TI - Assortative mating and differential male mating success in an ash hybrid zone population. AB - BACKGROUND: The structure and evolution of hybrid zones depend mainly on the relative importance of dispersal and local adaptation, and on the strength of assortative mating. Here, we study the influence of dispersal, temporal isolation, variability in phenotypic traits and parasite attacks on the male mating success of two parental species and hybrids by real-time pollen flow analysis. We focus on a hybrid zone population between the two closely related ash species Fraxinus excelsior L. (common ash) and F. angustifolia Vahl (narrow leaved ash), which is composed of individuals of the two species and several hybrid types. This population is structured by flowering time: the F. excelsior individuals flower later than the F. angustifolia individuals, and the hybrid types flower in-between. Hybrids are scattered throughout the population, suggesting favorable conditions for their local adaptation. We estimate jointly the best-fitting dispersal kernel, the differences in male fecundity due to variation in phenotypic traits and level of parasite attack, and the strength of assortative mating due to differences in flowering phenology. In addition, we assess the effect of accounting for genotyping error on these estimations. RESULTS: We detected a very high pollen immigration rate and a fat-tailed dispersal kernel, counter-balanced by slight phenological assortative mating and short-distance pollen dispersal. Early intermediate flowering hybrids, which had the highest male mating success, showed optimal sex allocation and increased selfing rates. We detected asymmetry of gene flow, with early flowering trees participating more as pollen donors than late flowering trees. CONCLUSION: This study provides striking evidence that long-distance gene flow alone is not sufficient to counter-act the effects of assortative mating and selfing. Phenological assortative mating and short-distance dispersal can create temporal and spatial structuring that appears to maintain this hybrid population. The asymmetry of gene flow, with higher fertility and increased selfing, can potentially confer a selective advantage to early flowering hybrids in the zone. In the event of climate change, hybridization may provide a means for F. angustifolia to further extend its range at the expense of F. excelsior. PMID- 17107612 TI - Economics methods in Cochrane systematic reviews of health promotion and public health related interventions. AB - BACKGROUND: Provision of evidence on costs alongside evidence on the effects of interventions can enhance the relevance of systematic reviews to decision-making. However, patterns of use of economics methods alongside systematic review remain unclear. Reviews of evidence on the effects of interventions are published by both the Cochrane and Campbell Collaborations. Although it is not a requirement that Cochrane or Campbell Reviews should consider economic aspects of interventions, many do. This study aims to explore and describe approaches to incorporating economics methods in a selection of Cochrane systematic reviews in the area of health promotion and public health, to help inform development of methodological guidance on economics for reviewers. METHODS: The Cochrane Database of Systematic Reviews was searched using a search strategy for potential economic evaluation studies. We included current Cochrane reviews and review protocols retrieved using the search that are also identified as relevant to health promotion or public health topics. A reviewer extracted data which describe the economics components of included reviews. Extracted data were summarised in tables and analysed qualitatively. RESULTS: Twenty-one completed Cochrane reviews and seven review protocols met inclusion criteria. None incorporate formal economic evaluation methods. Ten completed reviews explicitly aim to incorporate economics studies and data. There is a lack of transparent reporting of methods underpinning the incorporation of economics studies and data. Some reviews are likely to exclude useful economics studies and data due to a failure to incorporate search strategies tailored to the retrieval of such data or use of key specialist databases, and application of inclusion criteria designed for effectiveness studies. CONCLUSION: There is a need for consistency and transparency in the reporting and conduct of the economics components of Cochrane reviews, as well as regular dialogue between Cochrane reviewers and economists to develop increased capacity for economic analyses alongside such reviews. Use of applicable economics methods in Cochrane reviews can help provide the international context within which economics data can be interpreted and assessed as a preliminary to full economic evaluation. PMID- 17107613 TI - Social and demographic predictors of no transport prior to premature cardiac death: United States 1999-2000. AB - BACKGROUND: In the United States, over one-third of premature cardiac deaths occur outside of a hospital, without any transport prior to death. Transport prior to death is a strong, valid indicator of help-seeking behavior. We used national vital statistics data to examine social and demographic predictors of risk of no transport prior to cardiac death. We hypothesized that persons of lower social class, immigrants, non-metropolitan residents, racial/ethnic minorities, men, and younger decedents would be more likely to die prior to transport. METHODS: Our study population consisted of adult residents of the United States, aged 25 to 64 years, who died from heart disease during 1999-2000 (n = 242,406). We obtained transport status from the place of death variable on the death certificate. The independent effects of social and demographic predictor variables on the risk of a cardiac victim dying prior to transport vs. the risk of dying during or after transport to hospital were modeled using logistic regression. RESULTS: Results contradicted most of our a priori hypotheses. Persons of lower social class, immigrants, most non-metropolitan residents, and racial/ethnic minorities were all at lower risk of dying prior to transport. The greatest protective effect was found for racial/ethnic minority decedents compared with whites. The strongest adverse effect was found for marital status: the risk of dying with no transport was more than twice as high for those who were single (OR 2.35; 95% CI 2.29-2.40) or divorced (OR 2.29; 95% CI 2.24-2.34), compared with married decedents. Geographically, residents of the Western United States were at a 47% increased risk of dying prior to transport compared with residents of the metropolitan South. CONCLUSION: Our results suggest that marital status, a broad marker of household structure, social networks, and social support, is more important than social class or race/ethnicity as a predictor of access to emergency medical services for persons who suffer an acute cardiac event. Future research should focus on ascertaining "event histories" for all acute cardiac events that occur in a community, with the goal of identifying the residents most susceptible to cardiac fatalities prior to medical intervention and transport. PMID- 17107614 TI - Serendipitous identification of natural intergenotypic recombinants of hepatitis C in Ireland. AB - BACKGROUND: Recombination between hepatitis C single stranded RNA viruses is a rare event. Natural viable intragenotypic and intergenotypic recombinants between 1b-1a, 1a-1c and 2k-1b, 2i-6p, respectively, have been reported. Diagnostically recombinants represent an intriguing challenge. Hepatitis C genotype is defined by interrogation of the sequence composition of the 5' untranslated region [5'UTR]. Occasionally, ambiguous specimens require further investigation of the genome, usually by interrogation of the NS5B region. The original purpose of this study was to confirm the existence of a suspected mixed genotype infection of genotypes 2 and 4 by clonal analysis at the NS5B region of the genome in two specimens from two separate individuals. This initial identification of genotype was based on analysis of the 5'UTR of the genome by reverse line probe hybridisation [RLPH]. RESULTS: The original diagnosis of a mixed genotype infection was not confirmed by clonal analysis of the NS5B region of the genome. The phylogenetic analysis indicated that both specimens were natural intergenotypic recombinant forms of HCV. The recombination was between genotypes 2k and 1b for both specimens. The recombination break point was identified as occurring within the NS2 region of the genome. CONCLUSION: The viral recombinants identified here resemble the recombinant form originally identified in Russia. The RLPH pattern observed in this study may be a signature indicative of this particular type of intergenotype recombinant of hepatitis C meriting clonal analysis of NS2. PMID- 17107615 TI - Four-day antithrombin therapy does not seem to attenuate hypercoagulability in patients suffering from sepsis. AB - INTRODUCTION: Sepsis activates the coagulation system and frequently causes hypercoagulability, which is not detected by routine coagulation tests. A reliable method to evaluate hypercoagulability is thromboelastography (TEG), but this has not so far been used to investigate sepsis-induced hypercoagulability. Antithrombin (AT) in plasma of septic patients is decreased, and administration of AT may therefore reduce the acquired hypercoagulability. Not clear, however, is to what extent supraphysiologic plasma levels of AT decrease the acute hypercoagulability in septic patients. The present study investigates the coagulation profile of septic patients before and during four day high-dose AT therapy. METHODS: Patients with severe sepsis were randomly assigned to receive either 6,000 IU AT as a bolus infusion followed by a maintenance dose of 250 IU/hour over four days (n = 17) or placebo (n = 16). TEG, platelet count, plasma fibrinogen levels, prothrombin time and activated partial thromboplastin time were assessed at baseline and daily during AT therapy. RESULTS: TEG showed a hypercoagulability in both groups at baseline, which was neither reversed by bolus or by maintenance doses of AT. The hypercoagulability was mainly caused by increased plasma fibrinogen, and to a lesser extent by platelets. Plasmatic coagulation as assessed by the prothrombin time and activated partial thromboplastin time was similar in both groups, and did not change during the study period. CONCLUSION: The current study shows a distinct hypercoagulability in patients suffering from severe sepsis, which was not reversed by high-dose AT treatment over four days. This finding supports recent data showing that modulation of coagulatory activation in septic patients by AT does not occur before one week of therapy. PMID- 17107616 TI - A novel copper complex induces ROS generation in doxorubicin resistant Ehrlich ascitis carcinoma cells and increases activity of antioxidant enzymes in vital organs in vivo. AB - BACKGROUND: In search of a suitable GSH-depleting agent, a novel copper complex viz., copper N-(2-hydroxyacetophenone) glycinate (CuNG) has been synthesized, which was initially found to be a potential resistance modifying agent and later found to be an immunomodulator in mice model in different doses. The objective of the present work was to decipher the effect of CuNG on reactive oxygen species (ROS) generation and antioxidant enzymes in normal and doxorubicin-resistant Ehrlich ascites carcinoma (EAC/Dox)-bearing Swiss albino mice. METHODS: The effect of CuNG has been studied on ROS generation, multidrug resistance associated protein1 (MRP1) expression and on activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). RESULTS: CuNG increased ROS generation and reduced MRP1 expression in EAC/Dox cells while only temporarily depleted glutathione (GSH) within 2 h in heart, kidney, liver and lung of EAC/Dox bearing mice, which were restored within 24 h. The level of liver Cu was observed to be inversely proportional to the level of GSH. Moreover, CuNG modulated SOD, CAT and GPx in different organs and thereby reduced oxidative stress. Thus nontoxic dose of CuNG may be utilized to reduce MRP1 expression and thus sensitize EAC/Dox cells to standard chemotherapy. Moreover, CuNG modulated SOD, CAT and and GPx activities to reduce oxidative stress in some vital organs of EAC/Dox bearing mice. CuNG treatment also helped to recover liver and renal function in EAC/Dox bearing mice. CONCLUSION: Based on our studies, we conclude that CuNG may be a promising candidate to sensitize drug resistant cancers in the clinic. PMID- 17107617 TI - Systematic screening of polyphosphate (poly P) levels in yeast mutant cells reveals strong interdependence with primary metabolism. AB - BACKGROUND: Inorganic polyphosphate (poly P) occurs universally in all organisms from bacteria to man. It functions, for example, as a phosphate and energy store, and is involved in the activation and regulation of proteins. Despite its ubiquitous occurrence and important functions, it is unclear how poly P is synthesized or how poly P metabolism is regulated in higher eukaryotes. This work describes a systematic analysis of poly P levels in yeast knockout strains mutated in almost every non-essential gene. RESULTS: After three consecutive screens, 255 genes (almost 4% of the yeast genome) were found to be involved in the maintenance of normal poly P content. Many of these genes encoded proteins functioning in the cytoplasm, the vacuole or in transport and transcription. Besides reduced poly P content, many strains also exhibited reduced total phosphate content, showed altered ATP and glycogen levels and were disturbed in the secretion of acid phosphatase. CONCLUSION: Cellular energy and phosphate homeostasis is suggested to result from the equilibrium between poly P, ATP and free phosphate within the cell. Poly P serves as a buffer for both ATP and free phosphate levels and is, therefore, the least essential and consequently most variable component in this network. However, strains with reduced poly P levels are not only affected in their ATP and phosphate content, but also in other components that depend on ATP or free phosphate content, such as glycogen or secreted phosphatase activity. PMID- 17107618 TI - Tools for the identification of variable and potentially variable tandem repeats. AB - BACKGROUND: Tandem repeat arrays showing variation between sequences within a population, between strains or across species may have functional effects. The increasing availability of genomic sequence data makes routine description of observed variation possible, creating a need for tools to describe such variability. RESULTS: We present a set of programs that facilitate the identification of tandem repeats showing variation across multiple sequences or genomes, and the prediction of potentially polymorphic tandem repeats. The VNTRfinder (Variable Number of Tandem Repeats finder) program enables the detection of sequence length variation between arrays of inter-specific or intra specific tandem repeats. In the absence of comparable sequences to explore observed variation, predictions are provided describing which tandem repeats are more likely to be variable, to help guide and focus further experimental evaluation. CONCLUSION: These tools represent a resource for researchers interested in tandem repeats in nucleotide sequences that are most likely to be of clinical and evolutionary interest. The tools are available at http://bioinformatics.rcsi.ie/vntrfinder/. Downloadable versions for UNIX/LINUX and WINDOWS which permit the consideration of longer and more numerous sequences are also available. PMID- 17107619 TI - Household out-of-pocket payments for illness: evidence from Vietnam. AB - BACKGROUND: In Vietnam, illnesses create high out-of-pocket health care expenditures for households. In this study, the burden of illness in the Bavi district, Vietnam is measured based upon individual household health expenditures for communicable and non-communicable illnesses. The focus of the paper is on the relative effect of different illnesses on the total economic burden of health care on households in general and on households that have catastrophic health care spending in particular. METHODS: The study was performed by twelve monthly follow-up interviews of 621 randomly selected households. The households are part of the FilaBavi project sample--Health System Research Project. The heads of household were interviewed at monthly intervals from July 2001 to June 2002. RESULTS: For the population in the Bavi district, communicable illnesses predominate among the episodes of illness and are the reason for most household health care expenditure. This is the case for almost all groups within the study and for the study population as a whole. However, communicable illnesses are more dominant in the poor population compared to the rich population, and are more dominant in households that have very large, or catastrophic, health care expenditure, compared to those without such expenditures. CONCLUSION: The main findings indicate that catastrophic health care spending for a household is not usually the result of one single disastrous event, but rather a series of events and is related more to "every-day illnesses" in a developing country context than to more spectacular events such as injuries or heart illnesses. PMID- 17107621 TI - Prof-in-a-Box: using internet-videoconferencing to assist students in the gross anatomy laboratory. AB - BACKGROUND: The optimal learning environment for gross anatomy is the dissection laboratory. The Prof-in-a-Box (PiB) system has been developed where an anatomist using distance-learning technologies 'helps' students in a dissection laboratory at a different site. METHODS: The PiB system consists of: (1) an anatomist in his/her office with a computer and video camera; (2) a computer and 2 video cameras in the lab; (3) iChat AV software; (4) a secure server to host the PiB student 'consultation'. The PiB system allows the students and faculty to interact via audio and video providing an environment where questions can be asked and answered and anatomical structures can be identified 'at a distance' in real-time. The PiB system was set up at a prosected cadaver and made available for student use during 'office hours'. RESULTS: 25-30% of the students used the PiB system. Anatomical structures were identified, questions answered and demonstrations given 'at a distance' using the system. Students completed an optional questionnaire about the PiB system at the end of the semester. Results of the questionnaire indicate that the students were enthusiastic about the PiB system and wanted its use to be expanded in the future. CONCLUSION: Many of the functions of a faculty member in the gross anatomy dissection laboratory can be performed 'at a distance' using the PiB system. This suggests that a geographically dispersed faculty could assist in providing instruction in the dissection labs at multiple medical schools without needing to be physically present. PMID- 17107622 TI - The practice of physicians and nurses in the Brazilian Family Health Programme - evidences of change in the delivery health care model. AB - The article analyzes the practice of physicians and nurses working on the Family Health Programme (Programa de Saude da Familia or PSF, in Portuguese). A questionnaire was used to assess the evidences of assimilation of the new values and care principles proposed by the programme. The results showed that a great number of professionals seem to have incorporated the practice of home visits, health education actions and planning of the teams' work agenda to their routine labour activities. PMID- 17107620 TI - Short-wavelength sensitive opsin (SWS1) as a new marker for vertebrate phylogenetics. AB - BACKGROUND: Vertebrate SWS1 visual pigments mediate visual transduction in response to light at short wavelengths. Due to their importance in vision, SWS1 genes have been isolated from a surprisingly wide range of vertebrates, including lampreys, teleosts, amphibians, reptiles, birds, and mammals. The SWS1 genes exhibit many of the characteristics of genes typically targeted for phylogenetic analyses. This study investigates both the utility of SWS1 as a marker for inferring vertebrate phylogenetic relationships, and the characteristics of the gene that contribute to its phylogenetic utility. RESULTS: Phylogenetic analyses of vertebrate SWS1 genes produced topologies that were remarkably congruent with generally accepted hypotheses of vertebrate evolution at both higher and lower taxonomic levels. The few exceptions were generally associated with areas of poor taxonomic sampling, or relationships that have been difficult to resolve using other molecular markers. The SWS1 data set was characterized by a substantial amount of among-site rate variation, and a relatively unskewed substitution rate matrix, even when the data were partitioned into different codon sites and individual taxonomic groups. Although there were nucleotide biases in some groups at third positions, these biases were not convergent across different taxonomic groups. CONCLUSION: Our results suggest that SWS1 may be a good marker for vertebrate phylogenetics due to the variable yet consistent patterns of sequence evolution exhibited across fairly wide taxonomic groups. This may result from constraints imposed by the functional role of SWS1 pigments in visual transduction. PMID- 17107623 TI - Impaired pain sensation in mice lacking prokineticin 2. AB - Prokineticins (PKs), consisting of PK1 and PK2, are a pair of newly identified regulatory peptides. Two closely related G-protein coupled receptors, PKR1 and PKR2, mediate the signaling of PKs. PKs/PKRs participate in the regulation of diverse biological processes, ranging from development to adult physiology. A number of studies have indicated the involvement of PKs/PKRs in nociception. Here we show that PK2 is a sensitizer for nociception. Intraplantar injection of recombinant PK2 resulted in a strong and localized hyperalgesia with reduced thresholds to nociceptive stimuli. PK2 mobilizes calcium in dissociated dorsal root ganglion (DRG) neurons. Mice lacking the PK2 gene displayed strong reduction in nociception induced by thermal and chemical stimuli, including capsaicin. However, PK2 mutant mice showed no difference in inflammatory response to capsaicin. As the majority of PK2-responsive DRG neurons also expressed transient receptor potential vanilloid (TRPV1) and exhibited sensitivity to capsaicin, TRPV1 is likely a significant downstream molecule of PK2 signaling. Taken together, these results reveal that PK2 sensitize nociception without affecting inflammation. PMID- 17107624 TI - A correlated motif approach for finding short linear motifs from protein interaction networks. AB - BACKGROUND: An important class of interaction switches for biological circuits and disease pathways are short binding motifs. However, the biological experiments to find these binding motifs are often laborious and expensive. With the availability of protein interaction data, novel binding motifs can be discovered computationally: by applying standard motif extracting algorithms on protein sequence sets each interacting with either a common protein or a protein group with similar properties. The underlying assumption is that proteins with common interacting partners will share some common binding motifs. Although novel binding motifs have been discovered with such approach, it is not applicable if a protein interacts with very few other proteins or when prior knowledge of protein group is not available or erroneous. Experimental noise in input interaction data can further deteriorate the dismal performance of such approaches. RESULTS: We propose a novel approach of finding correlated short sequence motifs from protein protein interaction data to effectively circumvent the above-mentioned limitations. Correlated motifs are those motifs that consistently co-occur only in pairs of interacting protein sequences, and could possibly interact with each other directly or indirectly to mediate interactions. We adopted the (l, d)-motif model and formulate finding the correlated motifs as an (l, d)-motif pair finding problem. We present both an exact algorithm, D-MOTIF, as well as its approximation algorithm, D-STAR to solve this problem. Evaluation on extensive simulated data showed that our approach not only eliminated the need for any prior protein grouping, but is also more robust in extracting motifs from noisy interaction data. Application on two biological datasets (SH3 interaction network and TGFbeta signaling network) demonstrates that the approach can extract correlated motifs that correspond to actual interacting subsequences. CONCLUSION: The correlated motif approach outlined in this paper is able to find correlated linear motifs from sparse and noisy interaction data. This, in turn, will expedite the discovery of novel linear binding motifs, and facilitate the studies of biological pathways mediated by them. PMID- 17107626 TI - Comparison of PrASE and Pyrosequencing for SNP Genotyping. AB - BACKGROUND: There is an imperative need for SNP genotyping technologies that are cost-effective per sample with retained high accuracy, throughput and flexibility. We have developed a microarray-based technique and compared it to Pyrosequencing. In the protease-mediated allele-specific extension (PrASE), the protease constrains the elongation reaction and thus prevents incorrect nucleotide incorporation to mismatched 3'-termini primers. RESULTS: The assay is automated for 48 genotyping reactions in parallel followed by a tag-microarray detection system. A script automatically visualizes the results in cluster diagrams and assigns the genotypes. Ten polymorphic positions suggested as prothrombotic genetic variations were analyzed with Pyrosequencing and PrASE technologies in 442 samples and 99.8 % concordance was achieved. In addition to accuracy, the robustness and reproducibility of the technique has been investigated. CONCLUSION: The results of this study strongly indicate that the PrASE technology can offer significant improvements in terms of accuracy and robustness and thereof increased number of typeable SNPs. PMID- 17107625 TI - Ovarian epithelial cancer: a role for PGE2-synthesis and signalling in malignant transformation and progression. AB - BACKGROUND: The involvement of the cyclooxygenases (COX), in particular COX-2, is well documented for many tumours, e.g. colon, breast and prostate cancer, by both experimental and clinical studies. There are epidemiological data from subjects using NSAIDs, and experimental evidence supporting the hypothesis of prostaglandins (PGs) as regulators of tumourigenesis in the ovary. One of the end products of PG-synthesis, PGE2, regulates several key-processes, which are characteristic for tumour growth, e.g. angiogenesis, proliferation and apoptosisis. The present study investigated the pathway for PGE2-synthesis and signalling in ovarian tumourigenesis by analysing specimen from normal ovaries (n = 18), benign (B) (n = 8), borderline type (BL) (n = 6) and malignant tumours (AC) (n = 22). The expression and cell-specific localization of COX-1, COX-2, microsomal prostaglandin E synthase-1 (mPGES-1) and two of the receptors for PGE2, EP1 and EP2, were examined by immunoblotting (IB) and immunohistochemistry (IHC). RESULTS: The results are in line with earlier studies demonstrating an increase of COX-2 in AC compared to the normal ovary, B and BL tumours. Increased expressions were also observed for COX-1, mPGES-1 and EP-1 which all were significantly (p < 0.05) augmented in less differentiated AC (grades: moderately , poorly- and undifferentiated). The increase of COX-2 was also correlated to stage (FIGO classification) with significant elevations in stages II and III. EP1 was increased in stage III while no significant alterations were demonstrated for COX-1, mPGES-1 or EP2 for stage. IHC revealed staining of the tumour cells, but also increase of COX-1, COX-2, mPGES-1 and EP1-2 in the stromal compartment of AC (grades: moderately-, poorly- and undifferentiated). This observation suggests interactions between tumour cells and stromal cells (fibroblasts, immune cells), e.g. paracrine signalling mediated by growth factors, cytokines and possibly PGs. CONCLUSION: The increases of COX-1, COX-2, mPGES-1 and EP1-2 in epithelial ovarian cancer, supports the hypothesis that PGE2-synthesis and signalling are of importance for malignant transformation and progression. The observed augmentations of COX-1, COX-2 and mPGES-1 have implications for future therapeutic strategies. PMID- 17107627 TI - Powell's Pearls: Joe Vincent Meigs, MD (1892-1963). PMID- 17107628 TI - Gastrointestinal endoscopy in pregnant and lactating women: emerging standard of care to guide decision-making. AB - Endoscopic procedures generally are considered to be low-risk modalities, being performed on an on-demand basis in many ambulatory endoscopy centers and hospitals. However, the safety and efficacy of gastrointestinal endoscopy in pregnancy has not been well studied. In pregnancy, the risks to the fetus and mother are numerous and the magnitude of this risk will differ depending on trimester, the disease process being addressed, the underlying health and status of the maternofetal unit, and the nature of the endoscopic intervention being contemplated. Given the difficulties in performing prospective studies in the pregnant patient, these risks have not been well-quantified. Decision-making in practice is at times highly individualized. The obstetrician and endoscopist may therefore need to rely on prudent practice based on personal experience in conjunction with recognized underlying basic principles and consensus guidelines, rather than hard scientific data, to guide decision-making and advising patients. The purpose of this review is to make those caring for pregnant patients aware of the clinical considerations necessary to ensure a successful outcome and to provide guidance with respect to what is now considered to be the standard of care when managing pregnant and lactating patients in whom an endoscopic procedure is being contemplated. PMID- 17107629 TI - Preterm birth: a review of genetic factors and future directions for genetic study. AB - Preterm birth remains the leading cause of neonatal morbidity and mortality and is likely the result of interactions between specific genes and the maternal or fetal environment. The strong familial clustering of disease with documented increased risks in patients with a personal or family history of preterm birth and the racial disparities in the incidence of preterm birth support a genetic component of this condition. New technologies such as microarray, single nucleotide polymorphism analysis, and proteomics will lead to the eventual identification and characterization of the genetic etiology of preterm birth. PMID- 17107630 TI - Public health implications of cesarean on demand. AB - Cesarean rates have been rising in the United States. Recently, there has been an upsurge of interest in "cesarean on maternal request" in the absence of any medical indication, a phenomenon that will further increase the cesarean rate. This trend may not be benign on a population basis, and reliable data are lacking. This article reviews reasons for the increasing cesarean rate, describes maternal and neonatal consequences likely to accrue with a policy of cesarean on demand, and explores larger implications for public health. PMID- 17107631 TI - Refractory juvenile idiopathic arthritis: using autologous stem cell transplantation as a treatment strategy. AB - Cellular immune therapy for severe autoimmune diseases can now be considered when such patients are refractory to conventional treatment. The use of autologous stem cell transplantation (ASCT) to treat human autoimmune diseases has been initiated following promising results in a variety of animal models. Anecdotal observations have been made of autoimmune disease remission in patients who have undergone allogeneic bone marrow transplantation as a result of coincidental haematological malignancies. The possibility of inducing immunological self tolerance by ASCT is particularly attractive as a means for treating juvenile idiopathic arthritis (JIA). In this disease, ASCT restores self-tolerance both through a cell-intrinsic mechanism, involving the reprogramming of autoreactive T cells, and through a cell-extrinsic mechanism, involving a renewal of the immune balance between CD4+CD25+ regulatory T cells and other T cells. This review describes the clinical results of ASCT performed for this disease and the possible underlying immunological mechanisms. PMID- 17107632 TI - The effect of husbands' fertility preferences on couples' reproductive behaviour in rural Bangladesh. AB - Bangladesh society is profoundly gender stratified, and yet male roles in reproductive health processes have not been rigorously investigated. This study examines the association between men's reproductive health knowledge, attitude and behaviour and their wives' subsequent reproductive behaviour using longitudinal data from the International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B). A total of 4969 matched husband-and-wife data from a 1998 survey and women's contraceptive use history data following this survey are used. Results show a significant association between husbands' fertility preferences and current use of any family planning method. When wives' background characteristics, and husbands' background and socioeconomic characteristics are controlled for, the predicted probability of using a method of contraception among non-educated wives whose husbands want more children is 0.49 compared with 0.64 for those whose husbands do not want more children. However, the net effect of husbands' preference for additional children diminishes as wives' level of education increases. Among wives who had completed high school, the predicted probability of using a method of contraception is 0.70 for those whose husbands want more children compared with 0.69 for those whose husbands do not want more children. PMID- 17107633 TI - Levels and differentials in childhood mortality in South Africa, 1977-1998. AB - This study uses the 1996 Census and the 1998 Demographic and Health Survey (DHS) to investigate the level of and trend in infant and child mortality and their covariates in South Africa. Census estimates of childhood mortality are higher than those from the DHS. Analysis suggests that the former overestimate mortality while the latter are probably slightly too low. Both inquiries document a reversal of the trend toward lower mortality in the 1990s. Under-five mortality increased by about a third during the five years up to early 1998. By then the infant mortality rate was about 55 per 1000 and under-five mortality 72 per 1000. Other factors may explain the tapering off of the decline in mortality after the late 1980s but AIDS deaths account for its increase. Inequalities in childhood mortality between population groups, rooted in past discriminatory apartheid policies, shrank between the late-1970s and mid-1990s. However, they remain substantial and are largely unaccounted for by province, metropolitan residence and inter-group differences in mothers' education. The HIV/AIDS epidemic is likely to offset the beneficial impact of post-apartheid pro-poor policies and may exacerbate racial differences in childhood mortality in South Africa. There is an urgent need to improve the routine collection of statistics to monitor child mortality so as to assess progress towards the Millennium Development Goals and track inequalities. PMID- 17107635 TI - Children of homosexuals and transsexuals more apt to be homosexual: a reply to Cameron. PMID- 17107636 TI - Determinants of adolescent pregnancy in an urban area in Turkey: a population based case-control study. AB - The aim of this study was to determine the degree to which socioeconomic status is a risk factor for first birth at age 19 or younger in married women in an urban area of Turkey. The research was a population-based case-control study. The study group comprised all married and pregnant women aged 15-19 (adolescent pregnancies) attending primary care centres (144 subjects). Married women between 20 and 29 years of age, experiencing their first pregnancy (adult pregnancies), were determined as the control group (144 subjects). A questionnaire was completed for each subject during face-to-face interviews. Adolescent pregnancy was more frequent in women from families with a low socioeconomic status, as determined by occupation (class) and income; both were associated with adolescent pregnancy. Multiple logistic regression analysis identified seven factors associated with adolescent pregnancy: exposure to violence within the family prior to marriage; families partially opposed or unopposed to adolescent marriage; secondary school or lower education level; lack of social security; living in houses in which the number of persons per room was over 1; unemployed women; and having sisters with a history of adolescent pregnancy. PMID- 17107637 TI - Extending from PARs in Caenorhabditis elegans to homologues in Haemonchus contortus and other parasitic nematodes. AB - Signal transduction molecules play key roles in the regulation of developmental processes, such as morphogenesis, organogenesis and cell differentiation in all organisms. They are organized into 'pathways' that represent a coordinated network of cell-surface receptors and intracellular molecules, being involved in sensing environmental stimuli and transducing signals to regulate or modulate cellular processes, such as gene expression and cytoskeletal dynamics. A particularly important group of molecules implicated in the regulation of the cytoskeleton for the establishment and maintenance of cell polarity is the PAR proteins (derived from partition defective in asymmetric cell division). The present article reviews salient aspects of PAR proteins involved in the early embryonic development and morphogenesis of the free-living nematode Caenorhabditis elegans and some other organisms, with an emphasis on the molecule PAR-1. Recent advances in the knowledge and understanding of PAR-1 homologues from the economically important parasitic nematode, Haemonchus contortus, of small ruminants is summarized and discussed in the context of exploring avenues for future research in this area for parasitic nematodes. PMID- 17107638 TI - Feeding and circadian clocks. AB - The mammalian genome encodes at least a dozen of genes directly involved in the regulation of the feedback loops constituting the circadian clock. The circadian system is built up on a multitude of oscillators organized according to a hierarchical model in which neurons of the suprachiasmatic nuclei of the hypothalamus may drive the central circadian clock and all the other somatic cells may possess the molecular components allowing tissues and organs to constitute peripheral clocks. Suprachiasmatic neurons are driving the central circadian clock which is reset by lighting cues captured and integrated by the melanopsin cells of the retina and define the daily rhythms of locomotor activity and associated physiological regulatory pathways like feeding and metabolism. This central clock entrains peripheral clocks which can be synchronized by non photic environmental cues and uncoupled from the central one depending on the nature and the strength of the circadian signal. The human circadian clock and its functioning in central or peripheral tissues are currently being explored to increase the therapeutic efficacy of timed administration of drugs or radiation, and to offer better advice on lighting and meal timing useful for frequent travelers suffering from jet lag and for night workers' comfort. However, the molecular mechanism driving and coordinating the central and peripheral clocks through a wide range of synchronizers (lighting, feeding, physical or social activities) remains a mystery. PMID- 17107639 TI - Biodiversity of multiple Pregnancy-Associated Glycoprotein (PAG) family: gene cloning and chorionic protein purification in domestic and wild eutherians (Placentalia)--a review. AB - This review presents a broad overview of chorionic glycoproteins encoded by the Pregnancy-Associated Glycoprotein (PAG) gene family and also serves to illustrate how the recent discovery of the PAG family has contributed to our general knowledge of genome evolution, placental transcription and placental protein expression. The complex and large PAG family is restricted to the Artiodactyla order, although single PAG-like genes have also been identified in species outside the Artiodactyla. The PAGs are members of the aspartic proteinase (AP) superfamily. Unexpectedly, however, some members of the PAG family possess amino acid substitutions within and around the active site that likely render them unable to act as proteinases. This paper summarises the available information regarding biodiversity of PAG gene expression based on cDNA cloning, mRNA localisation studies and the structural organisation of the PAG genes with a particular emphasis on PAG promoters. It also compares available data regarding PAG protein purifications, sequencing and their N-glycodiversity. Finally, it discusses the scientific relevance, possible functional roles of the PAGs and describes possible profitable applications related to the detection of PAG proteins in the blood of pregnant domestic and wild species. PMID- 17107640 TI - Combined effect of salinomycin and feeding on whole body glucose kinetics in sheep fed a high-concentrate diet. AB - The aim of this study was to investigate the effects of salinomycin (SL) and feeding on whole body glucose kinetics in sheep fed a high-concentrate diet (25% orchardgrass hay and 75% commercial concentrate). Four adult sheep were fed the diet with or without 20 mg x kg(-1) diet of SL once daily for each 3 wk. The rates of glucose entry and utilization were determined before and during 3 h after feeding using a [ (13)C(6)] glucose dilution approach. Ruminal characteristics and concentrations of blood volatile fatty acids (VFA) and plasma glucose and insulin were also measured. Metabolizable energy intake was unaffected (P = 0.22) with SL. Salinomycin decreased (P = 0.06) the ratio of acetate to propionate in rumen fluid. Salinomycin increased (P = 0.01) both rates of entry and utilization of glucose, but did not affect (P > 0.10) concentrations of blood VFA or plasma glucose or insulin. Feeding caused gradual increases in concentrations of blood acetate (P < 0.01) and propionate (P = 0.01), a transient increase in plasma insulin concentration (P = 0.05), a transient decrease in plasma glucose concentration (P < 0.01), and persistent increases in both rates of glucose entry (P < 0.01) and utilization (P < 0.01). No SL x feeding interaction was observed (P > 0.10) on any measurements. We conclude that SL and feeding would have an additive effect on both rates of glucose entry and utilization without modifications with SL to feeding responses of peripheral concentrations of blood VFA, plasma glucose and insulin. PMID- 17107641 TI - The retina is more susceptible than the brain and the liver to the incorporation of trans isomers of DHA in rats consuming trans isomers of alpha-linolenic acid. AB - Trans polyunsaturated fatty acids are formed during heat treatments of vegetable oils from polyunsaturated fatty acids containing cis double bonds. After dietary intake, they are distributed in the body and are incorporated into nervous tissues including the retina. Since nervous tissues are known to be rich in n-3 fatty acids such as docosahexaenoic acid (DHA), we studied the ability of the retina and the brain to incorporate trans isomers of DHA formed in vivo from the dietary precursor trans alpha-linolenic acid. Wistar rats were fed with trans isomers of alpha-linolenic acid for 21 months. A linear incorporation of trans DHA and a decrease in cis DHA was observed in the retina, whereas no major changes were observed in the brain. In parallel to the modifications in retinal cis and trans DHA levels, the retinal functionality evaluated by the electroretinogram showed defects in animals that consumed trans alpha-linolenic acid. These results suggest that the mechanisms leading to the incorporation of cis and trans fatty acids are quite different in the retina when compared to the brain and the liver, the retina being more susceptible to changes in the dietary lipid contribution. PMID- 17107642 TI - Conception and development of a bibliographic database of blood nutrient fluxes across organs and tissues in ruminants: data gathering and management prior to meta-analysis. AB - In the organism, nutrient exchanges among tissues and organs are subject to numerous sources of physiological or nutritional variation, and the contribution of individual factors needs to be quantified before establishing general response laws. To achieve this, meta-analysis of data from publications is a useful tool. The objective of this work was to develop a bibliographic database of nutrient fluxes across organs and tissues of ruminant animals (Flora) under Access using the Merise method. The most important criteria for Flora were the ease to relate the various information, the exhaustivity and the accuracy of the data input, a complete description of the diets, taking into account the methods of the methodological procedures of measurement and analysis of blood nutrients and the traceability of the information. The conceptual data model was built in 6 parts. The first part describes the authors and source of publication, and the person in charge of data input. It clearly separates and identifies the experiments, the groups of animals and the treatments within a publication. The second part is concerned with feeds, diets and their chemical composition and nutritional value. The third and fourth parts describe the infusion of any substrates and the methods employed, respectively. The fifth part is devoted to the results of blood flows and nutrient fluxes. The sixth part gathers miscellaneous experimental information. All these parts are inter-connected. To model this database, the Merise method was utilised and 26 entities and 32 relationships were created. At the physical level, 93 tables were created, corresponding, for the majority, to entities and relationships of the data model. They were divided into reference tables (n = 65) and data tables (n = 28). Data processing was developed in Flora and included the control of the data, generic calculations of unknown data from given data, the automation of the estimation of the missing data or the chemical composition of the diets. It also included the construction of tables for meta analyses and the study of the variations of several factors within publications (pre-coding of meta-analyses). Overall, the system was built to facilitate the gathering, input, validation, management and retrieval of data from publications. PMID- 17107644 TI - MicroRNA involvement in mammary gland development and breast cancer. AB - MicroRNA (miRNA) are small non-coding RNA that post-transcriptionally regulate gene expression. In humans, miRNA genes may account for 2 to 3% of the total number of genes. Although the biological functions of most miRNA are unknown, their importance for development, cell proliferation, cell death, and morphogenesis has been demonstrated in several species. One could thus speculate that miRNA should be involved in the regulation of one of the organs that can undergo cycles of cell division, differentiation and dedifferentiation in the adult, the mammary gland. In this paper we summarise several reports dealing with the potential implication of miRNA in the mammary gland, most of them focussed on pathological situations, such as the appearance of breast cancer. These data suggest an implication of miRNA on mammary gland biology. However, direct evidence of this is still lacking. Expression profile analysis of miRNA during the normal mammary gland development could help in addressing this question and in identifying miRNA potentially involved. To this aim, we undertook such an analysis on mouse mammary gland at different stages (virgin, pregnancy, lactation and involution) and will present our preliminary results. PMID- 17107645 TI - Sulphated proteins secreted by rat mammary epithelial cells. AB - The main sulphated proteins secreted by rat mammary gland tissue have Mr of approximately 32 000, 27 000 and 25 000 Da. In addition, there are high Mr components which have a diffuse electrophoretic mobility (Mr > 200 000) and most likely corresponded to proteoglycans. The sulphate groups in the proteins with discrete Mr are most likely all linked to carbohydrates. These sulphated molecules were partially purified and identified to isoforms of rat alpha lactalbumin for the 25-27 kDa bands and to kappa-casein for the 32 kDa band. This pattern of protein sulphation is, as far as we know, quite specific to rat mammary epithelial cells. PMID- 17107646 TI - Translational efficiency of casein transcripts in the mammary tissue of lactating ruminants. AB - Caseins are essentially concentrated in the colloidal fraction of ruminant milks as highly hydrated and mineralized spherical particles, termed casein micelles. They form a group of four peptide chains (alpha(s1), beta, alpha(s2) and kappa), encoded by four structural genes (CSN1S1, CSN2, CSN1S2 and CSN3, respectively) of which the expression is regulated by lactogenic hormones. These phosphoproteins are synthesized, essentially during lactation, in the mammary epithelial cells and we show, for the first time, that their regulation is also controlled at the translational level. Apparently, the four casein messenger are not translated with the same efficiency. Specific amplification systems have been developed and optimized to quantify, by real time quantitative PCR (qPCR), transcripts encoding the four caseins starting from total RNA extracted from mammary tissues taken on goats (n = 4), ewes (n = 3) and cows (n = 3), in lactation. The relative proportions of each specific messenger (% of casein mRNA) were compared to the relative amounts of the corresponding caseins (% of whole casein) in milks sampled from the same animals, determined after fractionation by reverse phase HPLC and integration of the corresponding peak areas. From qPCR data, the four casein transcripts appeared to be present approximately at the same level of abundance (ca. 25%, except for defective genotypes at the CSN1S1 locus, in the goat) whereas the amounts of the corresponding proteins in milk were ranging between 9 and 38% of the whole casein fraction. A comparison of specific translational efficiencies (% of protein in milk/% of transcript in the mammary tissue), showed that alpha(s1)- and beta-casein transcripts are translated ca. 3- to 4-fold more efficiently than alpha(s2)- and kappa-casein transcripts. This seems to be the rule in the three ruminant species studied. More or less optimal contexts for initiation of translation (Kozak recognition sequence of the start codon) as well as 3' untranslated region (UTR) sequences and length might explain, at least in part, our results. These preliminary results which have now to be confirmed with a larger number of individuals to strengthen our findings and conclusions, provides, however, a rational explanation to the unbalanced casein distribution (approximate proportions 4:1:4:1 for alpha(s1):alpha (s2):beta:kappa, respectively) reported for ruminant milks. The possible effects of specific secondary structures in the 5' and 3' UTRs of casein messengers still have to be considered. PMID- 17107647 TI - Preparation of recombinant proteins in milk to improve human and animal health. AB - Milk is a very abundant source of proteins for animal and human consumption. Milk composition can be modified using transgenesis, including exogenous gene addition and endogenous gene inactivation. The study of milk protein genes has provided researchers with regulatory regions capable of efficiently and specifically driving the expression of foreign genes in milk. The projects underway are aimed at modifying milk composition, improving its nutritional value, reducing mammary infections, providing consumers with antipathogen proteins and preparing purified recombinant proteins for pharmaceutical use. The present paper summarises the current progress in this field. PMID- 17107648 TI - Changes in mammary uptake and metabolic fate of glucose with once-daily milking and feed restriction in dairy cows. AB - The aim of this review is to better understand the regulation of milk yield in response to once-daily milking and feed restriction. Glucose is the principal precursor for the synthesis of lactose (a major osmotic agent in milk), and participates in determining the milk volume produced. When applying these two breeding factors, reductions in milk yield are associated with a reduction in milk lactose yield and in the arterial flow of glucose, due to a decrease in the mammary blood flow. The ability of the udder to extract glucose is altered with once-daily milking but not necessarily with feed restriction. Lactose synthesis is down-regulated in response to once-daily milking and feed restriction but the percentage of the extracted glucose which is converted into lactose is differently affected in response to treatments. No marked change is observed with once daily milking whereas this would be increased with feed restriction and in contrast, depressed with fasting. PMID- 17107649 TI - 15-month-old infants detect violations in pretend scenarios. AB - Are 15-month-old infants able to detect a violation in the consistency of an event sequence that involves pretense? In Experiment 1, infants detected a violation when an actor pretended to pour liquid into one cup and then pretended to drink from another cup. In Experiment 2, infants no longer detected a violation when the cups were replaced with objects not typically used in the context of drinking actions, either shoes or tubes. Experiment 3 showed that infants' difficulty in Experiment 2 was not due to the use of atypical objects per se, but arose from the novelty of seeing an actor appearing to drink from these objects. After receiving a single familiarization trial in which they observed the actor pretend to drink from either a shoe or a tube, infants now detected a violation when the actor pretended to pour into and to drink from different shoes or tubes. Thus, at an age (or just before the age) when infants are beginning to engage in pretend play, they are able to show comprehension of at least one aspect of pretense in a violation-of-expectation task: specifically, they are able to detect violations in the consistency of pretend action sequences. PMID- 17107650 TI - Calibration in isothermal titration calorimetry: heat and cell volume from heat of dilution of NaCl(aq). AB - An isothermal titration calorimeter of the perfusion type (MicroCal model VP-ITC) is calibrated using the heat of dilution of NaCl in water. The relative apparent molar enthalpy function (L(phi)) for NaCl(aq) varies strongly and nonlinearly with concentration in the low-concentration region (<0.2M) that is sampled easily and extensively in a single program of injections of NaCl solution into water. This nonlinearity makes it possible to calibrate with respect to two quantities: the measured heat and the active cell volume. The heat factor is determined with typical standard error 0.003; its value in the current case is 0.987. The cell volume factor is 0.93 but is quite sensitive to possible systematic errors in the temperature and in the literature values for L(phi). Both correction factors are closely tied to the delivered volume from the injection syringe, which required a correction factor of 0.973, attributed to an instrumental gear ratio error. Temperature calibration of the instrument showed a small offset of 0.12K at the temperature 25 degrees C of the experiments, but the error increased to more than 1K at 46 degrees C. The experiments were not able to distinguish clearly between mixing algorithms that assume instantaneous mixing on injection and those that assume instantaneous injection followed by mixing; however, examination of these algorithms has revealed an error in a program widely used to analyze isothermal titration calorimetry data. PMID- 17107651 TI - Escherichia coli DNA polymerase III epsilon subunit increases Moloney murine leukemia virus reverse transcriptase fidelity and accuracy of RT-PCR procedures. AB - In an effort to improve reverse transcriptase (RT) fidelity, we measured the error rate of Moloney murine leukemia virus (MMLV) RT in the presence of several autonomous and DNA polymerase-associated 3'-5' exonucleases using a lacZ forward mutation assay. A number of 3'-5' exonucleases were found to lower the error rate of MMLV RT, including p53, Escherichia coli DNA polymerase III epsilon subunit, and the proofreading activities associated with T4, varphi29, and E. coli pol I DNA polymerases. The bacterial epsilon subunit increased RNA-dependent DNA synthesis fidelity by approximately threefold and was the only 3'-5' exonuclease tested that did not deleteriously affect RT-PCR yields. Further testing showed that RT-PCR mutant frequencies were reduced significantly by performing cDNA synthesis in the presence of epsilon subunit, followed by PCR with a high fidelity proofreading DNA polymerase. DNA sequence analysis was used to show that the combination of MMLV RT/epsilon subunit and PfuUltra DNA polymerase produces approximately eightfold fewer errors compared with the commonly used combination of MMLV RT and a Taq-based high-fidelity blend, consistent with predictions based on experimentally determined polymerase error rates. PMID- 17107652 TI - Development of fast Fourier transformation continuous cyclic voltammetry as a highly sensitive detection system for ultra trace monitoring of penicillin V. AB - Fast continuous cyclic voltammetry was used as a detection method for penicillin V in a flow injection system. Additionally, a special computer-based numerical calculation method (using fast Fourier transformation) providing enhancement of the analyte signal and noise reduction is introduced here. During the measurements, the potential waveform (consisting of the potential steps of cleaning, stripping, and the potential ramp) was continuously applied to an Au disk microelectrode (12.5 microm in radius). In particular, the effects of accumulation potential, sweep rate, and delay time on the sensitivity and selectivity of the method were investigated. Eventually, the stripping time was found to be less than 300 ms, the detection limit of the method was 7.0x10(-12) M, and the associated relative standard deviation at 5.0x10(-6) M penicillin V was 2.3 for eight runs. PMID- 17107653 TI - Development of an HPLC assay for Staphylococcus aureus sortase: evidence for the formation of the kinetically competent acyl enzyme intermediate. AB - Many bacterial surface proteins containing an LPXTG motif are anchored to the cell wall peptidoglycan by catalysis with the thiol transpeptidase sortase. The transpeptidation and hydrolysis reactions of sortase have been proposed to proceed through a common acyl enzyme intermediate. The reactions of Staphylococcus aureus sortase with fluorogenic substrate Abz-LPETG-Dnp in the presence or absence of triglycine were characterized in this study to gain additional insight into the kinetic mechanism of sortase. We report here the development of a reverse-phase HPLC assay to identify and characterize sortase reaction intermediates. The HPLC results provide for the first time clear evidence for the formation of a kinetically competent acyl enzyme intermediate during the overall transpeptidation reaction. The results also suggest that sortase undergoes an unexpected intramolecular acyl transfer reaction in the absence of a nucleophile. The significance of this type of HPLC assay as a tool to study enzyme mechanism is discussed. PMID- 17107654 TI - Effects of training to implement new tools and working methods to reduce knee load in floor layers. AB - The purpose was to measure the effect of a participatory ergonomics implementation strategy consisting of information, education, and facilitation on the use of new tools and working methods in the floor laying trade. Floor layers (n=292) were trained in using new working methods. The effects were evaluated by using questionnaires, interviews, and assessments of quality and productivity. Following the training, 43% had used the new working methods weekly/daily compared to 11% before. There was a reduction in the degree of self-reported pain in the knees among the floor layers using the new working methods weekly or daily compared to those using them never or occasionally. The musculoskeletal complaints did not increase from any other region and the quality and the productivity of the work were not decreased. This indicates that within a 3 months perspective the implementation strategy succeeded. PMID- 17107655 TI - The effect of cyclic deformation and solute binding on solute transport in cartilage. AB - Diffusive transport must play an important role in transporting nutrients into cartilage due to its avascular nature. Recent theoretical studies generally support the idea that cyclic loading enhances large molecule transport through advection. However, to date, reactive transport, i.e. the effects of solute binding, has not yet been taken into consideration in cyclically deformed cartilage. In the present study, we develop a reactive transport model to describe the potential role of binding of solute within cyclically deformed cartilage. Our results show that binding does have a significant effect on transport, particularly for the low IGF-I concentrations typical of synovial fluid. A dynamic loading regime of high strain magnitudes (up to 10%) in combination with high frequencies (e.g. 1 Hz) was seen to produce the most dramatic results with enhanced total uptake ratio as high as 25% averaged over the first 5h of cyclic loading. PMID- 17107657 TI - Attentional bias and general orienting processes in bipolar disorder. AB - Attentional biases for emotional stimuli and general orienting processes were examined in bipolar disorder, using a modified dot-probe task with a spatial cueing paradigm incorporated in it. Bipolar patients in a euthymic state (i.e., remission), bipolar patients in a mildly depressed state, and non-psychiatric controls participated. General orienting results showed that within the patient group as a whole, measures of depressed mood were positively associated with a relative inability to disengage attention. Attentional bias results showed that bipolar patients in a mildly depressed state, in comparison with controls, directed their attention away from depression-related words and positive words. The bias away from positive words was equally present in both patient groups and part of a trait effect, demonstrated by the comparison of patients in a euthymic state and controls. The bias away from depression-related words was mood state dependent and within the patient group as a whole correlated negatively with measures of depressed mood. It is proposed that biases for emotional stimuli are related to the transition of mood states, characteristic for bipolar disorder. PMID- 17107656 TI - Regulation and induction of CYP3A11, CYP3A13 and CYP3A25 in C57BL/6J mouse liver. AB - This study reports that dexamethasone (DEX) significantly induces CYP3A11, CYP3A13 and CYP3A25 mRNA expression in male and female 4 days, 3 weeks and 18 weeks old C57BL/6J mice. Furthermore, CYP3A activity, as measured by erythromycin N-demethylation, is also significantly increased. PXR, RXRalpha and CAR are known to be involved in the induction of CYP3As. Here we report nuclear receptors PXR and RXRalpha but not CAR demonstrate gender- and age-dependent expression. Also, treatment of C57BL/6J mice with DEX induces PXR but not RXRalpha or CAR. In summary, we demonstrate DEX is not only able to up-regulate CYP3A expression and activity, but also the nuclear receptor PXR through which it may exert this effect. Furthermore, the gender- and age-dependent pattern of basal PXR and RXRalpha expression is similar to the 3 CYP3As analysed. PMID- 17107658 TI - Studies on the role of HtpG in the tetrapyrrole biosynthesis pathway of the cyanobacterium Synechococcus elongatus PCC 7942. AB - In cyanobacterium Synechococcus elongatus PCC 7942, we observed that htpG overexpression caused remarkable growth inhibition. In addition, subcellular fractionation experiments showed that HtpG was localized in the membrane fraction. To understand its function in cyanobacteria, we carried out yeast two hybrid screening to identify specific proteins interacting with HtpG, and found out, HemE, uroporphyrinogen decarboxylase. When compared to the wild-type strain, the htpG-null mutant and -overexpressing strains exhibited higher and lower cytosolic HemE activity, based on the coproporphyrin production, respectively. These results strongly suggest that HtpG is involved in the regulation of tetrapyrrole biosynthesis through interacting with HemE protein. PMID- 17107659 TI - The differentiation of mesenchymal stem cells by mechanical stress or/and co culture system. AB - Differentiation of mesenchymal stem cells (MSCs) into anterior cruciate ligament (ACL) cells is regulated by many factors. Mechanical stress affects the healing and remodeling process of ACL after surgery in important ways. Besides, co culture system had also showed the promise to induce MSCs toward different kinds of cells on current research. The purpose of this study was to investigate the gene expression of ACL cells' major extracellular matrix (ECM) component molecules of MSCs under three induction groups. In addition, to follow our previous study, cell electrophoresis technique and mRNA level gene expression of MSC protein were also used to analyze the differentiation of MSCs. The results reveal that specific regulatory signals which released from ACL cells appear to be responsible for supporting the selective differentiation toward ligament cells in co-culture system and mechanical stress promotes the secretion of key ligament ECM components. Therefore, the combined regulation could assist the development of healing and remolding of ACL tissue engineering. Furthermore, this study also verifies that cell electrophoresis could be used in investigation of cell differentiation. Importantly, analysis of the data suggests the feasibility of utilizing MSCs in clinical applications for repairing or regenerating ACL tissue. PMID- 17107660 TI - LNA-modified oligonucleotides effectively drive intramolecular-stable hairpin to intermolecular-duplex state. AB - Sequence-specific hybridization of antisense and antigene agent to the target nucleic acid is an important therapeutic strategy to modulate gene expression. However, efficiency of such agents falls due to inherent intramolecular-secondary structures present in the target that pose competition to intermolecular hybridization by complementary antisense/antigene agent. Performance of these agents can be improved by employing structurally modified complementary oligonucleotides that efficiently hybridize to the target and force it to transit from an intramolecular-structured-state to an intermolecular-duplex state. In this study, the potential of variably substituted locked nucleic acid-modified oligonucleotides (8mer) to hybridize and disrupt highly stable, secondary structure of nucleic acid has been biophysically characterized and compared with the conventionally used unmodified DNA oligonucleotides. The target here is a stem-loop hairpin oligonucleotide-a structure commonly present in most structured nucleic acids and known to exhibit an array of biological functions. Using fluorescence-based studies and EMSA we prove that LNA-modified oligonucleotides hybridize to the target hairpin with higher binding affinity even at lower concentration and subsequently, force it to assume a duplex conformation. LNA modified oligonucleotides may thus, prove as potential therapeutic candidates to manipulate gene expression by disruption of biologically relevant nucleic acid secondary structure. PMID- 17107661 TI - Evolution of the syntrophic interaction between Desulfovibrio vulgaris and Methanosarcina barkeri: Involvement of an ancient horizontal gene transfer. AB - The sulfate reducing bacteria Desulfovibrio vulgaris and the methanogenic archaea Methanosarcina barkeri can grow syntrophically on lactate. In this study, a set of three closely located genes, DVU2103, DVU2104, and DVU2108 of D. vulgaris, was found to be up-regulated 2- to 4-fold following the lifestyle shift from syntroph to sulfate reducer; moreover, none of the genes in this gene set were differentially regulated when comparing gene expression from various D. vulgaris pure culture experiments. Although exact function of this gene set is unknown, the results suggest that it may play roles related to the lifestyle change of D. vulgaris from syntroph to sulfate reducer. This hypothesis is further supported by phylogenomic analyses showing that homologies of this gene set were only narrowly present in several groups of bacteria, most of which are restricted to a syntrophic lifestyle, such as Pelobacter carbinolicus, Syntrophobacter fumaroxidans, Syntrophomonas wolfei, and Syntrophus aciditrophicus. Phylogenetic analysis showed that all three individual genes in the gene set tended to be clustered with their homologies from archaeal genera, and they were rooted on archaeal species in the phylogenetic trees, suggesting that they were horizontally transferred from archaeal methanogens. In addition, no significant bias in codon and amino acid usages was detected between these genes and the rest of the D. vulgaris genome, suggesting the gene transfer may have occurred early in the evolutionary history so that sufficient time has elapsed to allow an adaptation to the codon and amino acid usages of D. vulgaris. This report provides novel insights into the origin and evolution of bacterial genes linked to the lifestyle change of D. vulgaris from a syntrophic to a sulfate-reducing lifestyle. PMID- 17107662 TI - Cardiac mitochondrial connexin 43 regulates apoptosis. AB - Connexin 43 (Cx43) is thought to be present largely in the plasma membrane and its function solely to provide low resistance electrical connection between myocytes. A recent report suggested the presence of Cx43 in the mitochondria as well. We confirmed the presence of Cx43 in the mitochondria isolated from adult rat ventricles with the Cx43 immunoreactivity fractionating to the outer mitochondrial membrane. Mitochondrial Cx43 is mostly phosphorylated only detected by a phospho-specific antibody. Using a Ca2+ -sensitive electrode and Western blot, we showed that the gap junction inhibitors 18-beta-glycyrrhetinic acid (beta-GA), oleamide, and heptanol all induced concomitant release of Ca2+ and cytochrome C in isolated mitochondria whereas the inactive analog 18-beta glycyrrhizic acid failed to do so. In low density neonatal myocyte culture with no appreciable cell-cell contacts, beta-GA induced apoptosis as assessed by TUNEL staining. Our results suggest a novel role of Cx43 as a regulator of mitochondrial physiology and myocyte apoptosis. PMID- 17107663 TI - Long term low-dose arsenic exposure induces loss of DNA methylation. AB - Arsenic ranks as the number one toxic environmental contaminant. In humans, arsenic exposure is associated with various forms of cancer, cardiovascular and skin diseases, neuropathies of the central nervous system, and genotoxic and immunotoxic effects. Although a well recognized human carcinogen, arsenic itself is not a potent mutagen and has been thought to act through epigenetic mechanisms that modify DNA methylation patterns, perhaps in conjunction with DNA-damaging agents. To develop preliminary support for a more thorough examination of this hypothesis, we have measured the effect of submicromolar and low-micromolar concentrations of arsenite on the methylation status of DNA and the biochemical reactions that regulate it. We find that arsenic causes the depletion of S adenosylmethionine, the main cellular methyl donor, and represses the expression of the DNA methyltransferase genes DNMT1 and DNMT3A. Possibly as a consequence of these two complementary mechanisms, long-term exposure to arsenic results in DNA hypomethylation. PMID- 17107664 TI - Glypicans are differentially expressed during patterning and neurogenesis of early mouse brain. AB - Glypicans are essential modulators of cell signalling during embryogenesis. Little is known about their functions in brain development. We show here that mouse glypicans (gpc-1 to gpc-6) are differentially expressed in embryonic brains during key morphogenetic events. In gastrulating embryos, gpc-4 is the only glypican expressed in anterior visceral endoderm. During neural tube closure, gpc 4 transcripts are restricted to the anterior neural ridge and telencephalon. At this stage, gpc-1 expression shifts from trunk and head mesenchyme to neural tube. Gpc-3 mRNA appears across the ventral neural tube, then in the lamina terminalis and hypothalamus. Gpc-2 and gpc-6 transcripts are in all brain compartments. Gpc-5 is found in ventral brains as neurogenesis starts. Onset of neurogenesis also coincides with differential expression of glypican genes either in neural progenitors or in differentiating neurons. The novel expression sites of glypicans shown here contribute to the identification of signalling molecules involved in brain patterning. PMID- 17107665 TI - Homer1c interacts with Hippi and protects striatal neurons from apoptosis. AB - By the two hybrid screening of mouse brain cDNA library, we identified Hppi, a cell death-promoting protein, as a binding partner of postsynaptic scaffold protein Homer1c. Hippi interacted specifically with Homer1c but not with its homologue Homer2. It was reported that Hippi, when complexed with Hip1, induces the apoptosis in striatal neurons and may cause Huntington's disease. We found that this apoptotic effect of Hippi was specific to the striatum and was not observed in hippocampal neurons. Furthermore, the apoptotic effect of Hippi was prevented when Homer1c was co-expressed in cultured striatal neurons. The protective effect of Homer1c was diminished when Hippi binding domain was deleted. These results suggest that Homer1c may play an important role in the mechanisms of neuronal death in the striatum. PMID- 17107666 TI - All-trans retinoic acid negatively regulates cytotoxic activities of nature killer cell line 92. AB - NK cells are key components of innate immune systems and their activities are regulated by cytokines and hormones. All-trans retinoic acid (ATRA), as a metabolite of vitamin A and an immunomodulatory hormone, plays an important role in regulating immune responses. In the present study, we investigated the effect of ATRA on human NK cell line NK92. We found that ATRA dose-dependently suppressed cytotoxic activities of NK92 cells without affecting their proliferation. To explore the mechanisms underlying the ATRA influence on NK92 cells, we examined the production of cytokines (TNF-alpha, IFN-gamma), gene expression of cytotoxic-associated molecules (perforin, granzyme B, nature killer receptors (NCRs), and NKG2D), and the activation of NF-kappaB pathways related with immune response. Our results demonstrated that ATRA suppressed NF-kappaB activity and prevented IkappaBalpha degradation in a dose-dependent way, inhibited IFN-gamma production and gene expression of granzyme B and NKp46. Our findings suggest that ATRA is a negative regulator of NK92 cell activation and may act as a potential regulator of anti-inflammatory functions in vivo. PMID- 17107667 TI - Epidermal growth factor up-regulates transforming growth factor-beta receptor type II in human dermal fibroblasts via p38 mitogen-activated protein kinase pathway. AB - TGF-beta receptors (TbetaRs) are serine/threonine kinase receptors that bind to TGF-beta and propagate intracellular signaling through Smad proteins. TbetaRs are repressed in some human cancers and expressed at high levels in several fibrotic diseases. We demonstrated that epidermal growth factor (EGF) up-regulates type II TGF-beta receptor (TbetaRII) expression in human dermal fibroblasts. EGF-mediated induction of TbetaRII expression was inhibited by the treatment of fibroblasts with a specific p38 mitogen-activated protein kinase (MAPK) inhibitor, SB203580, whereas MEK inhibitor PD98059 did not block the up-regulation of TbetaRII by EGF. EGF induced the TbetaRII promoter activity, and this induction was significantly blocked by SB203580, but not by PD98059. The overexpression of the dominant negative form of p38alpha or p38beta significantly reduced the induction of TbetaRII promoter activity by EGF. These results indicate that the EGF-mediated induction of TbetaRII expression involves the p38 MAPK signaling pathway. The EGF mediated induction of TbetaRII expression may participate in a synergistic interplay between EGF and TGF-beta signaling pathway. PMID- 17107668 TI - Ca2+ -dependent splicing of neurexin IIalpha. AB - Neurexins are synaptic adhesion proteins encoded by 3 genes (NRXN1, NRXN2, and NRXN3) each transcribed from 2 promoters to yield longer (alpha) and shorter (beta) forms. The primary gene transcripts undergo extensive alternative splicing leading to products that may differ in synaptic coupling properties. Here we show that depolarization of neurons modulates splicing of NRXN2alpha, particularly at splice sites 1 and 3. Furthermore, we demonstrate that exclusion of exon 11 at splice site 3 is calcium-dependent. These data indicate neuronal activity dependent splicing of NRXN2alpha. This dynamic process may be important for maintenance of mature neuronal circuits. PMID- 17107669 TI - PUMA is critical for neonatal cardiomyocyte apoptosis induced by endoplasmic reticulum stress. AB - OBJECTIVE: Puma (p53-upregulated modulator of apoptosis), a proapoptotic BH3-only member of the Bcl-2 protein family, has been implicated in the pathomechanism of several diseases, including cancer, AIDS, and ischemic brain disease. We have recently shown that Puma is required for cardiac cell death upon ischemia/reperfusion of mouse hearts. Since ischemia/reperfusion is also associated with endoplasmic reticulum (ER) stress, in the present study we investigated whether Puma contributes to the ER stress-dependent component of cardiomyocyte apoptosis. METHODS: Primary cultures of rat and mouse neonatal cardiomyocytes were treated with 3 muM thapsigargin or 100 ng mL(-1) tunicamycin. Puma levels were suppressed by adenoviral delivery of shRNA or targeted deletion of the puma gene. Puma expression was detected by RT-PCR and Western blotting. Apoptosis was assessed by TUNEL assay, caspase-3 cleavage, and cytochrome c release. RESULTS: We have shown that in rat neonatal cardiac myocytes, thapsigargin or tunicamycin treatment led to ER-stress, transcriptional upregulation of Puma, and apoptosis. Most importantly, cardiac myocytes acquired resistance to ER stress-induced apoptosis if Puma expression was downregulated by adenoviral delivery of shRNA or eliminated by targeted deletion in knockout mice. CONCLUSION: Taken together, our data indicate that Puma is a critical component of ER stress-induced apoptosis in cardiac myocytes, and inhibition of Puma activity may be used to treat cardiac infarcts or prevent heart failure by blocking ER stress-induced apoptosis. PMID- 17107670 TI - CaMKII-mediated increased lusitropic responses to beta-adrenoreceptor stimulation in ANP-receptor deficient mice. AB - OBJECTIVE: Mice with genetic disruption of the guanylyl cyclase-A (GC-A) receptor for atrial natriuretic peptide (ANP), have chronic arterial hypertension and marked cardiac hypertrophy. Intriguingly, despite pronounced remodeling, cardiac contractile functions and cardiomyocyte Ca(2+)-handling are preserved and even enhanced. The present study aimed to characterize the specific molecular mechanisms preventing cardiac failure. METHODS AND RESULTS: Contractile function and expression as well as phosphorylation of regulatory proteins were evaluated in isolated perfused working hearts from wild-type and GC-A KO mice under baseline conditions and during beta(1)-adrenergic stimulation. Ca(i)(2+) transients were monitored in Indo-1 loaded isolated adult cardiomyocytes. Cardiac contractile, especially lusitropic responsiveness to beta-adrenergic stimulation was significantly increased in GC-A KO mice. This was concomitant to enhanced expression and activation of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII), increased dual-site phosphorylation of phospholamban (PLB) at Ser(16) and Thr(17), enhanced amplitude of Ca(i)(2+) transients, and accelerated Ca(i)(2+) decay. In contrast, the expression of cardiac ryanodine receptors and phosphorylation at Ser(2809) and Ser(2815) was not altered. Pharmacological inhibition of CaMKII-but not of protein kinase A-mediated PLB phosphorylation totally abolished the increased effects of beta-adrenergic stimulation on cardiac contractility and Ca(i)(2+)-handling. Thus, acceleration of sarcoplasmic reticulum Ca(2+)-uptake and increased availability of Ca(2+) for contraction, both secondary to increased CaMKII-mediated PLB phosphorylation, seem to mediate the augmented responsiveness of GC-A KO hearts to catecholamines. CONCLUSION: Our observations show that increased CaMKII activity enhances the contractile relaxation response of hypertrophic GC-A KO hearts to beta-adrenergic stimulation and emphasize the critical role of CaMKII-dependent pathways in beta(1) adrenoreceptor modulation of myocardial Ca(2+)-homeostasis and contractility. PMID- 17107671 TI - Modulation of cholestasis-induced antinociception in rats by two NMDA receptor antagonists: MK-801 and magnesium sulfate. AB - Acute cholestasis is associated with increased activity of the endogenous opioid system that results to changes including analgesia. N-methyl-d-aspartate (NMDA) receptors are involved in the nociceptive pathway and play a major role in the development of morphine induced analgesia. The magnesium acts as a non competitive NMDA receptor antagonist by blocking the NMDA receptor channel. Considering the reported antinociceptive effect of magnesium sulfate as a NMDA receptor antagonist and the existence of close functional links between NMDA receptor antagonists and magnesium with the opioid system, we studied the effect of acute and chronic administration of MK-801 as a NMDA antagonist and magnesium sulfate on modulation of nociception in an experimental model of elevated endogenous opioid tone, acute cholestasis, using the tail-flick paradigm. Cholestasis was induced by ligation of the main bile duct using two ligatures and then transsection of the duct at the midpoint between them. A significant increase (P<0.001) in nociception threshold was observed in bile duct ligated rats compared to unoperated and sham-operated animals. In acute treatment, MK-801 (0.1 mg/kg, b.i.d), but not magnesium (150 mg/kg magnesium sulfate, i.e. 30 mg/kg of Mg(+2), i.p., b.i.d.) increased antinociception in cholestatic rats compared to saline treated cholestatics (P<0.05). In chronic treatment, administration of MK-801 or magnesium sulfate for 7 consecutive days, increased tail-flick latency (P<0.05, P<0.01) in cholestatic animals compared to saline treated cholestatics. These data showed that NMDA receptor pathway is involved in modulation of cholestasis-induced antinociception in rats and that repeated dosages of magnesium sulfate similar to MK-801 is able to modulate nociception in cholestasis. PMID- 17107672 TI - In vitro electrophysiological mechanisms for antiarrhythmic efficacy of resveratrol, a red wine antioxidant. AB - Resveratrol (trans-3, 4', 5-trihydroxystilbene), a natural antioxidant derived from grapes, has beneficial effects against coronary heart disease. Its electrophysiological characteristics for antiarrhythmic efficacy are largely unknown; thus, this study aims to explore the resveratrol's antiarrhythmic effects and conduction system in isolated hearts as well as its electrophysiological effects on cardiac myocytes. In the experiment, resveratrol suppressed the ischemia/reperfusion-induced ventricular arrhythmias in Langendorff-perfused rat hearts. In the current clamp study of the experiment, resveratrol prolonged the action potential duration (APD(50) and APD(90)) and suppressed the upstroke velocity of the action potential (V(max)). In the voltage clamp study, resveratrol inhibited sodium inward current (I(Na)) in a concentration-dependent manner and negative-shifted the voltage-dependent inactivation curve. Resveratrol also reduced the calcium inward current (I(Ca), 51.2+/-13.3% at 100 microM). Furthermore, the transient (I(to)) and sustained (I(ss)) outward potassium currents were decreased 60.2+/-5.7% and 42.3+/-5.2% after exposure to resveratrol (100 microM), respectively. The inward rectifier potassium current (I(K1)) was also reduced 24.2+/-7.0% in the presence of resveratrol (100 microM). In the isolated heart perfusion model, resveratrol (100 microM) prolonged AV nodal refractory period, the Wenckebach cycle length and the conduction through AV node and His-Purkinje system. In conclusion, resveratrol increased the cardiac effective refractory period mainly through inhibiting the ionic channels including I(Na), I(to) and I(ss) which could contribute to the conversion of ischemia/reperfusion-induced lethal arrhythmias. PMID- 17107673 TI - NMR investigation of DNA primer-template models: structural insights into dislocation mutagenesis in DNA replication. AB - Slipped frameshift intermediates can occur when DNA polymerase slows or stalls at sites of DNA lesions. However, this phenomenon is much less common when unmodified DNA is replicated. In order to study the effect of templating bases on the alignment of primer-templates, NMR structural investigation has been performed on primer-template oligonucleotide models which mimic the situation that dNTP has just been incorporated opposite template. NMR evidence reveals the occurrence of misalignment when dGTP is incorporated opposite template T with a downstream nucleotide C. Depending on the template sequence, further extension of the primer can lead to realignment. PMID- 17107674 TI - Lhca5 interaction with plant photosystem I. AB - In the outer antenna (LHCI) of higher plant photosystem I (PSI) four abundantly expressed light-harvesting protein of photosystem I (Lhca)-type proteins are organized in two heterodimeric domains (Lhca1/Lhca4 and Lhca2/Lhca3). Our cross linking studies on PSI-LHCI preparations from wildtype Arabidopsis and pea plants indicate an exclusive interaction of the rarely expressed Lhca5 light-harvesting protein with LHCI in the Lhca2/Lhca3-site. In PSI particles with an altered LHCI composition Lhca5 assembles in the Lhca1/Lhca4 site, partly as a homodimer. This flexibility indicates a binding-competitive model for the LHCI assembly in plants regulated by molecular interactions of the Lhca proteins with the PSI core. PMID- 17107675 TI - In vitro effects of homologous natriuretic peptides on growth hormone and prolactin release in the tilapia, Oreochromis mossambicus. AB - C-type natriuretic peptide (CNP) cDNA was cloned from the tilapia brain and its inferred mature sequence was chemically synthesized together with previously cloned tilapia A-type and B-type natriuretic peptides (ANP and BNP). The cloned CNP belongs to the CNP-1 type of teleosts. Reverse-transcription polymerase chain reaction showed that the ANP and BNP genes were hardly expressed in the tilapia brain and pituitary, whereas the CNP gene was expressed strongly in the brain and slightly in the pituitary. Effects of homologous natriuretic peptides (100 nM each) on growth hormone (GH) and prolactin (PRL) release were examined using dispersed tilapia pituitary cells. Tilapia ANP and BNP stimulated GH and PRL release during 4-8, and 8-24 h of incubation. BNP appeared to be more potent than ANP, also stimulating GH and PRL release during 0-4 h of incubation. CNP stimulated GH release only during 4-8 h of incubation; CNP was without effect on PRL release. All three NPs stimulated GH and PRL mRNA expression in dispersed pituitary cells following 24 h of incubation. ANP and BNP significantly elevated intracellular cGMP accumulation in dispersed pituitary cells after 15 min of exposure, whereas no effect of CNP was observed. These results indicate a long lasting stimulation of GH and PRL release by ANP and BNP that is mediated, at least in part, by the guanylyl cyclase-linked NP receptor. PMID- 17107676 TI - Leishmania major metacaspase can replace yeast metacaspase in programmed cell death and has arginine-specific cysteine peptidase activity. AB - The human protozoan parasite Leishmania major has been shown to exhibit several morphological and biochemical features characteristic of a cell death program when differentiating into infectious stages and under a variety of stress conditions. Although some caspase-like peptidase activity has been reported in dying parasites, no caspase gene is present in the genome. However, a single metacaspase gene is present in L. major whose encoded protein harbors the predicted secondary structure and the catalytic dyad histidine/cysteine described for caspases and other metacaspases identified in plants and yeast. The Saccharomyces cerevisiae metacaspase YCA1 has been implicated in the death of aging cells, cells defective in some biological functions, and cells exposed to different environmental stresses. In this study, we describe the functional heterologous complementation of a S. cerevisiae yca1 null mutant with the L. major metacaspase (LmjMCA) in cell death induced by oxidative stress. We show that LmjMCA is involved in yeast cell death, similar to YCA1, and that this function depends on its catalytic activity. LmjMCA was found to be auto-processed as occurs for caspases, however LmjMCA did not exhibit any activity with caspase substrates. In contrast and similarly to Arabidopsis thaliana metacaspases, LmjMCA was active towards substrates with arginine in the P1 position, with the activity being abolished following H147A and C202A catalytic site mutations. These results suggest that metacaspases are members of a family of peptidases with a role in cell death conserved in evolution notwithstanding possible differences in their catalytic activity. PMID- 17107677 TI - Gastrointestinal nematode parasites of sheep: a dynamic model for their effect on liveweight gain. AB - This paper presents an individual-based model for gastrointestinal nematode parasites of sheep and includes the effect of these parasites on the liveweight performance of young sheep. Parasitism is known to affect the host animal in at least two ways. The first induces a loss of appetite in the host, which reduces pasture consumption compared with the parasite-free animal. This effect is examined in the first part of the study. The second major effect of parasitism is a reduction in the metabolic efficiency of the host which decreases nutrients available for maintenance and growth. The latter part of the paper examines the consequences of incorporating this effect on the liveweight changes in individuals in a group of sheep. Previous models addressing this issue have only given mean liveweight and worm burden changes. PMID- 17107678 TI - Sexual differences in larval life history traits of acanthocephalan cystacanths. AB - Sexual differences in life history traits, such as size dimorphism, presumably arise via sexual selection and are most readily observed in adults. For complex life-cycle parasites, however, sexual selection may also have consequences for larval traits, e.g., growth in intermediate hosts. Two acanthocephalan species (Acanthocephalus lucii and Echinorhynchus borealis) were studied to determine, whether larval life histories differ between males and females. The size of female A. lucii cystacanths had a much stronger relationship with intermediate host size than males, suggesting females invest more in growth and are consequently more limited by resources. No relationship between host size and cystacanth size was observed for E. borealis. For both species, female cystacanths survived longer in a culture medium composed entirely of salts, which could suggest that females have greater energy reserves than males. A comparative analysis across acanthocephalan species indicated that sexual size dimorphism at the adult stage correlates with cystacanth dimorphism. However, the relationship was not isometric; cystacanths do not reach the same level of sexual dimorphism as adults, possibly due to resource constraints. Our results suggest that larval life histories diverge between males and females in some acanthocephalans, and this is seemingly a consequence of sexual selection acting on adults. PMID- 17107679 TI - Lipoprotein distribution and biological variation of 24S- and 27 hydroxycholesterol in healthy volunteers. AB - 24S- and 27-hydroxycholesterol are obligatory intermediates of cholesterol catabolism and play an important role in the maintenance of whole-body cholesterol homeostasis. Using an HPLC-MS method for oxysterol quantification, the distribution of esterified and unesterified oxysterols in lipoprotein subfractions as well as the influence of daytime, food intake and menstrual cycle on oxysterol concentrations were investigated in healthy volunteers. Moreover, reference intervals for 24S- and 27-hydroxycholesterol in plasma as well as the corresponding levels for 27-hydroxycholesterol in the HDL subfraction were established in 100 healthy volunteers. Both circulating oxysterols are mainly transported in association with HDL and LDL--primarily in the esterified form. No significant diurnal changes and no variations during menstrual cycle of either absolute or cholesterol-related plasma levels were detected. In contrast to 24S hydroxycholesterol in plasma and 27-hydroxycholesterol in the HDL subfraction, the 95% reference intervals of 27-hydroxycholesterol both in plasma and the non HDL subfraction were higher in males than in females. The concentrations of 27 hydroxycholesterol in plasma and the non-HDL subfraction showed strong positive correlations with the concentrations of cholesterol, non-HDL cholesterol and triglycerides. Our data on the lipoprotein distribution of oxysterols as well as on their intra- and inter-individual variation set the stage for future clinical studies. PMID- 17107680 TI - Co-current and counter-current imbibition in independent tubes of non axisymmetric geometry. AB - Experiments that illustrate and quantify the basics of co- and counter-current spontaneous imbibition have been conducted in a series of simple model pore systems. The fundamental pore geometry is a rod in an angled round-bottomed slot with the rod touching a capping glass plate. The capillaries thus formed by the surfaces of the slot, rod and plate do not have circular cross-sections but more complicated geometric structures with angular corners. The tubes formed at each side of the rod connect at both ends. A viscous, refined oil was applied from one end. For co-current experiments, the opposite end was left open to the atmosphere and oil imbibed into both tubes. For counter-current experiments the opposite end was sealed and connected to a sensitive pressure transducer. Oil imbibed into the smaller capillary and expelled air as a series of bubbles from the end of the larger capillary. Bubble snap-off was observed to be rate-dependent and occurred at a lower curvature than that of the cylindrical meniscus that just fits inside the tube. Only the corners of the larger capillary filled with oil during counter current imbibition. Meniscus curvatures were calculated using the Mayer and Stowe Princen method and were compared with actual values by measuring the capillary rise in the tubes; agreement was close. A simple model for co-current and counter current imbibition has also been developed and the predictions compared with the experimental results. The model results were in agreement with the experiments. The experiments demonstrate that the capillary back pressure generated by the interfaces and bubbles in counter-current imbibition can slow the process significantly. PMID- 17107681 TI - Unusually large acrylamide induced effect on the droplet size in AOT/Brij30 water in-oil microemulsions. AB - Droplet microemulsions are widely used as templates for controlled synthesis of nanometer sized polymer gel beads for use as, e.g., nanobiosensors. Here we examine water-in-oil microemulsions typically used for preparation of sensors. The cores of the microemulsion droplets are constituted by an aqueous component consisting of water, reagent monomer mixture, buffer salts, and the relevant dyes and/or enzymes. The cores are encapsulated by a mixture of the surfactants Brij30 and AOT and the resulting microemulsion droplets are suspended in a continuous hexane phase. The size of the final polymer particles may be of great importance for the applications of the sensors. Our initial working hypothesis was that the size of the droplet cores and therefore the size of the synthesized polymer gel beads could be controlled by the surfactant-to-water ratio of the template microemulsion. In the present work we have tested this hypothesis and investigated how the monomers and the ratio between the two surfactants affect the size of the microemulsion droplets and the microemulsion domain. We find that the monomers in water have a profound effect on the microemulsion domain as well as on the size of the microemulsion droplets. The relation between microemulsion composition and droplet size is in this case more complicated than assumed in standard descriptions of microemulsions [R. Strey, Colloid Polym. Sci. 272 (1994) 1005-1019; I. Danielsson, B. Lindman, Colloids Surf. 3 (1981) 391-392; Y. Chevalier, T. Zemb, Rep. Progr. Phys. 53 (1990) 279-371]. PMID- 17107682 TI - Lignocellulosic fiber charge enhancement by catalytic oxidation during oxygen delignification. AB - A series of one-stage oxygen delignification treatments with a softwood (SW) kraft pulp were studied employing 0.0-0.5% of a bismuth ruthenium pyrochlore oxide catalyst. The results demonstrated that a 0.09-0.18% charge of catalyst in an oxygen stage provided a 52.2-116.0% increase of carboxylic acid groups in the cellulosic component of kraft pulps without a significant decrease in fiber viscosity. A 3-factor at 3-level (L(9)3(3)) orthogonal experimental design was used to identify the main factors influencing acid group formation in pulp carbohydrates. The relative significance of experimental parameters for polysaccharide acid group formation was the molar equivalent NaOH, oxygen pressure, and finally, reaction temperature under the experimental conditions studied. The optimized reaction parameters for fiber charge development were shown to be 85-100 degrees C, 2.5% NaOH, and 800-960 kPa oxygen pressure. Pulps with higher fiber carboxylic acid content introduced by catalytic oxidation during oxygen delignification yielded a 10.9-33.7% increase in fiber charge after elemental chlorine free (ECF) pulp bleaching. The enhanced fiber charge resulted in 6.7-17.1% increase in paper sheet tensile index at comparable pulp viscosity. PMID- 17107683 TI - Study of the surface energy of montmorillonite using PACHA formalism. AB - The surface energy of exchanged forms of montmorillonite has been computed using a partial charge model introduced in the calculation of the lattice energy. The results are in correct agreement with data on heats of adsorption. However, the difference between theoretical and experimental data increases from light cations to heavy cations associated with the clay network. This indicates that the heat of adsorption depends highly on the step hydration of the cations. This type of behavior was expected and thus the quantification seems realistic. PMID- 17107684 TI - Effect of terbium(III) chloride on the micellization properties of sodium decyl- and dodecyl-sulfate solutions. AB - The effect of TbCl3 on the aggregation processes of the anionic surfactants sodium decyl sulfate (SDeS) and sodium dodecyl sulfate (SDS) has been investigated. Electrical conductivity data, combined with Tb(III) luminescence measurements suggest that the formation of micelles involving TbCl3 and SDS occurs at concentrations below the critical micelle concentration (cmc) of the pure surfactants; the formation of these mixed aggregates was also monitored by light scattering, which indicates that the addition of TbCl3 to surfactant concentration at values below the pure surfactant cmc results in a much greater light scattering than that found with pure sodium alkylsulfate surfactant micelles. This phenomenon is dependent upon the alkyl chain length of the surfactant. With Tb(III)/DS-, complexes are formed with a cation/anion binding ratio varying from 3 to 6, which depends upon the initial concentration of Tb(III). This suggests that the majority of the cation hydration water molecules can be exchanged by the anionic surfactant. When the carbon chain length decreases, interactions between surfactant and Tb(III) also decrease, alterations in conductivity and fluorescence data are not so significant and, consequently, no binding ratio can be detected even if existing. The surfactant micellization is dependent on the presence of electrolyte in solution with apparent cmc being lower than the corresponding cmc value of pure SDS. PMID- 17107685 TI - Quantitative detection of therapeutic proteins and their metabolites in serum using antibody-coupled ProteinChip Arrays and SELDI-TOF-MS. AB - One of the important steps in developing protein therapeutics is the determination of their preliminary PK in vivo. These data are essential to design optimal dosing in animal models prior to progressing to clinical trials in man. The quantitative detection of protein therapeutics in serum is traditionally performed by ELISA, which has the prerequisite of the availability of the appropriate monoclonal antibodies. We have developed an alternative method using polyclonal antibodies immobilized on ProteinChip Arrays and SELDI-TOF mass spectrometry. This method has an advantage over ELISA since it provides simultaneously information on the clearance rate of the protein and it's in vivo processing. We compared these two methods using a RANTES variant, [(44)AANA(47)] RANTES as the test protein in this study. Using SELDI-TOF mass spectrometry, we were able to establish that the protein is readily oxidized in serum, and moreover is processed in vivo to produce a truncated 3-68 protein, and undergoes a further cleavage to produce the 4-68 protein. These modifications are not identified by ELISA, whilst the serum exposure profiles determined by the two methods show essentially similar protein concentration values. PMID- 17107686 TI - Prediction of CD4(+) T cell epitopes restricted to HLA-DP4 molecules. AB - We have set up a method to predict peptide binding to HLA-DP4 molecules. These HLA II molecules are the most frequent worldwide and hence are an interesting target for epitope-based vaccines. The prediction is based on quantitative matrices built with binding data for peptides substituted at anchoring positions for HLA-DP4. A set of 98 peptides of various origins was used to compare the prediction with binding activity. At different prediction thresholds, the positive predictive value and the sensitivity of the prediction ranged from 50% to 80%, demonstrating its efficiency. This prediction method can be applied to the entire genomes of pathogens and large peptide sequences derived from tumor antigens. PMID- 17107687 TI - Preparation of Au coated polystyrene beads and their application in an immunoassay. AB - A novel immunoassay method based on polystyrene beads coated with Au nanoparticles (Au@PS) is described. Au nanoparticles were prepared by reductive reaction, and then deposited on the surface of polystyrene beads to form Au coatings. Results indicated that the Au coatings had good stability and that human IgG was immobilized at a concentration of 16 microg/g Au@PS. FITC-labeled rabbit-anti-human IgG and FITC-labeled rabbit-anti-goat IgG were employed to react with the human IgG on Au@PS. Fluorescence imaging results showed that the reaction had good immuno-specificity. In addition, further experiments at the single-bead level indicated that the linear range was 0.05-15 microg/ml, and that the FITC signal could be detected even when the target antibody concentration was as low as 0.01 microg/ml. The assay results were compared with an enzyme-linked immunosorbent assay (ELISA), and showed relatively good reliability. PMID- 17107688 TI - Crystal structure of archaeal photolyase from Sulfolobus tokodaii with two FAD molecules: implication of a novel light-harvesting cofactor. AB - UV exposure of DNA molecules induces serious DNA lesions. The cyclobutane pyrimidine dimer (CPD) photolyase repairs CPD-type - lesions by using the energy of visible light. Two chromophores for different roles have been found in this enzyme family; one catalyzes the CPD repair reaction and the other works as an antenna pigment that harvests photon energy. The catalytic cofactor of all known photolyases is FAD, whereas several light-harvesting cofactors are found. Currently, 5,10-methenyltetrahydrofolate (MTHF), 8-hydroxy-5-deaza-riboflavin (8 HDF) and FMN are the known light-harvesting cofactors, and some photolyases lack the chromophore. Three crystal structures of photolyases from Escherichia coli (Ec-photolyase), Anacystis nidulans (An-photolyase), and Thermus thermophilus (Tt photolyase) have been determined; however, no archaeal photolyase structure is available. A similarity search of archaeal genomic data indicated the presence of a homologous gene, ST0889, on Sulfolobus tokodaii strain7. An enzymatic assay reveals that ST0889 encodes photolyase from S. tokodaii (St-photolyase). We have determined the crystal structure of the St-photolyase protein to confirm its structural features and to investigate the mechanism of the archaeal DNA repair system with light energy. The crystal structure of the St-photolyase is superimposed very well on the three known photolyases including the catalytic cofactor FAD. Surprisingly, another FAD molecule is found at the position of the light-harvesting cofactor. This second FAD molecule is well accommodated in the crystal structure, suggesting that FAD works as a novel light-harvesting cofactor of photolyase. In addition, two of the four CPD recognition residues in the crystal structure of An-photolyase are not found in St-photolyase, which might utilize a different mechanism to recognize the CPD from that of An-photolyase. PMID- 17107690 TI - Ecology of player-to-player contact in boys' youth soccer play. AB - INTRODUCTION: Youth soccer (football) injuries occur for a wide range of reasons, but the most frequent cause of injury is via player-to-player contact. This study was designed to study the ecology of collisions between players during youth soccer play. METHOD: Six teams of 11- and 12-year-old male players were followed over the course of a full season. Games were videotaped and reviewed to address three primary questions: how frequently do player-to-player collisions occur; when and where on the field do those collisions occur; and what is the rate of falls and injuries as a result of player-to-player contact. RESULTS: A total of 1,279 player-to-player collisions was observed, or an average of 65.59 collisions per game. Nearly half of the observed collisions resulted in one or both players falling to the ground, and about one-tenth resulted in the referee calling a foul, but very few of the collisions (less than 1%) resulted in an injury. Collisions occurred relatively consistently throughout the games, no matter what the score was. They occurred most frequently in the midfield area, when the ball was on or near the ground, and when players were attempting to retrieve a loose, uncontrolled ball. CONCLUSIONS: Results are discussed with respect to implications for injury prevention. PMID- 17107689 TI - Rates of DSM-IV psychiatric disorders among adolescents in a large metropolitan area. AB - We present prevalence data for adolescents in a large metropolitan area in the US and the association of DSM-IV diagnoses to functional impairment and selected demographic correlates. We sampled 4175 youths aged 11-17 years from households enrolled in large health maintenance organizations. Data were collected using questionnaires and the Diagnostic Interview Schedule for Children, Version IV (DISC-IV). Impairment was measured using the Child Global Assessment Scale and diagnostic specific impairment in the DISC-IV. 17.1% of the sample met DSM-IV criteria for one or more disorders in the past year; 11% when only DISC impairment was considered and 5.3% only using the CGAS. The most prevalent disorders were anxiety (6.9%), disruptive (6.5%), and substance use (5.3%) disorders. The most prevalent specific disorders were agoraphobia, conduct and marijuana abuse/dependence, then alcohol use and oppositional defiant disorder. Younger youths and females had lower odds for any disorder, as did youths from two parent homes. There was increased odds associated with lower family income. Females had greater odds of mood and anxiety disorders, males of disruptive and substance use disorders. There were greater odds of mood and disruptive disorders for older youths. Prevalences were highly comparable to recent studies using similar methods in diverse non-metropolitan populations. We found associations with age, gender, and to a lesser extent, socioeconomic status reported in previous studies. The inclusion of both diagnosis-specific impairment and global impairment reduced prevalence rates significantly. Our results suggest commonality of prevalences and associated factors in diverse study settings, including urban and rural areas. PMID- 17107691 TI - Roles of nuclear receptors in the up-regulation of hepatic cholesterol 7alpha hydroxylase by cholestyramine in rats. AB - Nuclear receptors are involved in regulating the expression of cholesterol 7alpha hydroxylase (CYP7A1), however, their roles in the up-regulation of CYP7A1 by cholestyramine (CSR) are still unclear. In the present study, male Wistar rats were divided into four groups and fed [high sucrose + 10% lard diet] (H), [H + 3% CSR diet] (H + CSR), [H + 0.5% cholesterol + 0.25% sodium cholate diet] (C), or [C + 3% CSR diet] (C + CSR) for 2 weeks. Cholestyramine decreased serum and liver cholesterol levels significantly in rats fed C-based diets, but had no effect on these parameters in rats fed H-based diets. Cholestyramine raised hepatic levels of CYP7A1 mRNA and activity in both groups. The gene expression of hepatic ATP binding cassettes A1 and G5, regulated by liver X receptor (LXR), were unchanged and down-regulated by cholestyramine, respectively. The mRNA levels of the hepatic ATP-binding cassette B11 and short heterodimer partner (SHP), regulated by farnesoid X receptor (FXR), were not changed by cholestyramine. C-based diets, which contained cholesterol and cholic acid, increased SHP mRNA levels compared to H-based diets. Consequently, in rats fed the C+CSR diet, hepatic FXR was activated by dietary bile acids, but the hepatic CYP7A1 mRNA level was increased 16-fold compared to that in rats fed an H diet. These results suggest that cholestyramine up-regulates the expression of CYP7A1 independently via LXR- or FXR-mediated pathways in rats. PMID- 17107692 TI - Assessment of contamination by polychlorinated biphenyls and aliphatic and aromatic hydrocarbons in sediments of the Santos and Sao Vicente Estuary System, Sao Paulo, Brazil. PMID- 17107693 TI - Acute effects of an alcoholic drink on food intake: aperitif versus co-ingestion. AB - To compare the effects on appetite and energy intake (EI) within a meal and across 4 days of drinking wine either before (aperitif) or with (co-ingestion) a meal, 11 men attended the laboratory on three occasions. On each occasion participants were given breakfast in the laboratory then 3 h later returned for a two course (garlic bread, pizza) lunch, then recorded food intake for the remainder of the day and the next 3 days. In the control condition, participants ate lunch ad libitum; in the aperitif condition 375 ml of red wine was consumed 20 min before lunch; and in the co-ingestion condition 125 ml of red wine was consumed with the starter and 250 ml of red wine was consumed with the main course. Subjective ratings of appetite and mood were administered before and after the meal. EI at lunch was greater when wine was consumed (p<0.01) (aperitif: 6436+/-435 kJ; co-ingestion: 6254+/-417 kJ) compared to control (5125+/-262 kJ). In particular, intake of the starter was enhanced by wine consumption. Having wine as an aperitif significantly influenced subjective ratings of light headedness and fatigue. These results suggest that the effects of wine on appetite are immediate, and stimulate food intake early in the meal. Total EI during the test days was significantly higher than during subsequent days revealing a tendency to overeat in the laboratory, exacerbated by drinking wine before or with lunch. However, lean healthy men adjusted total EI on the days following the laboratory in all conditions. PMID- 17107694 TI - Propagation of spike and wave activity to the medial prefrontal cortex and dorsal raphe nucleus of WAG/Rij rats. AB - Although there is pharmacological evidence for the involvement of the serotonergic system in the expression of spike and wave discharges (SWDs) in experimental absence epilepsy, no direct investigation of this paroxysm in the dorsal raphe nucleus (DRN), one of the main serotonergic nuclei, has been carried out. We have now recorded the EEG simultaneously with local field potentials and unit activity in DRN from WAG/Rij rats, one of the best established models of absence epilepsy during spontaneous SWDs. We have also compared this activity to that in the thalamocortical networks, where SWDs are generated, and in the medial prefrontal cortex (mPFC), as this brain area is reciprocally connected to the DRN. We have found that SWDs propagate to the DRN with a short delay, and that the firing rate of its neurons changes during this type of paroxysm. These results provide the first direct evidence for clear alterations in the firing properties of mPFC and DRN neurons during spontaneous SWDs. PMID- 17107695 TI - Cardiac regulation in the socially monogamous prairie vole. AB - Social experiences, both positive and negative, may influence cardiovascular regulation. Prairie voles (Microtus ochrogaster) are socially monogamous rodents that form social bonds similar to those seen in primates, and this species may provide a useful model for investigating neural and social regulation of cardiac function. Cardiac regulation has not been studied previously in the prairie vole. Radiotelemetry transmitters were implanted into adult female prairie voles under anesthesia, and electrocardiographic parameters were recorded. Autonomic blockade was performed using atenolol (8 mg/kg ip) and atropine methyl nitrate (4 mg/kg ip). Several variables were evaluated, including heart rate (HR), HR variability and the amplitude of respiratory sinus arrhythmia. Sympathetic blockade significantly reduced HR. Parasympathetic blockade significantly increased HR, and reduced HR variability and the amplitude of respiratory sinus arrhythmia. Combined autonomic blockade significantly increased HR, and reduced HR variability and respiratory sinus arrhythmia amplitude. The data indicate that autonomic function in prairie voles shares similarities with primates, with a predominant vagal influence on cardiac regulation. The current results provide a foundation for studying neural and social regulation of cardiac function during different behavioral states in this socially monogamous rodent model. PMID- 17107696 TI - Platelets in equine recurrent airway obstruction. AB - Platelets contribute to the pathogenesis of human allergic airway disease. The aim of this study was to compare platelet activating factor (PAF)-induced platelet aggregation and thromboxane (Tx) production, plasma Tx and 5 hydroxytryptamine (5-HT) in ponies with recurrent airway obstruction (RAO), an hypersensitivity to inhaled antigens, and normal ponies, before and after antigen exposure. Plasma 5-HT was significantly higher in ponies with RAO but was not further increased by antigen challenge. There was no difference between PAF induced platelet aggregation or Tx production, or in plasma Tx before or after challenge. These data suggest there may be a difference between platelet 5-HT uptake in RAO and normal ponies but do not provide evidence of platelet activation following antigen exposure. PMID- 17107697 TI - Pharmacokinetics of enrofloxacin in turkeys. AB - The pharmacokinetics of enrofloxacin (EFL) and its active metabolite ciprofloxacin (CIP) was investigated in 7-8 month old turkeys (6 birds per sex). EFL was administered intravenously (i.v.) and orally (p.o.) at a dose 10 mg kg( 1) body weight. Blood was taken prior to and at 0.17, 0.33, 0.5, 1, 2, 3, 4, 6, 8, 10 and 24 h following drug administration. The concentrations of EFL and CIP in blood serum were determined by high-performance liquid chromatography (HPLC). Serum concentrations versus time were analysed by a noncompartmental analysis. The elimination half-live and the mean residence time of EFL after i.v. injection for the serum were after oral administration 6.64+/-0.90 h, 8.96+/-1.18 h and 6.92+/-0.97 h, 11.91+/-1.87 h, respectively. After single p.o. administration, EFL was absorbed slowly (MAT=2.76+/-0.48 h) with time to reach maximum serum concentrations of 6.33+/-2.54 h. Maximum serum concentrations was 1.23+/-0.30 microg mL(-1). Oral bioavailability for for EFL after oral administration was found to be 69.20+/-1.49%. The ratios C(max)/MIC and AUC(0 --> 24)/MIC were respectively from 161.23+/-5.9 h to 12.90+/-0.5 h for the pharmacodynamic predictor C(max)/MIC, and from 2153.44+/-66.6 h to 137.82+/-4.27 h for AUC(0 --> 24)/MIC, for the different clinically significant microorganisms, whose values for MIC varies from 0.008 microg L(-1) to 0.125 microg mL(-1). PMID- 17107698 TI - A situation in which a local nontoxic refuge promotes pest resistance to toxic crops. AB - In order to delay the development of pest resistance to genetically engineered insecticidal crop varieties, it is current practice to grow "refugees" of non toxic plants close to insecticidal crops. We model such a toxic/nontoxic crop complex as an open system with a small stream of toxin-susceptible immigrants. We find that, for intermediate values of the dominance of a pest gene for resistance to the toxin, the local refuge can spoil the benefit that is provided by the immigrant stream. We provide formulas for some important boundaries in parameter space. PMID- 17107699 TI - Love wave propagation in functionally graded piezoelectric material layer. AB - An exact approach is used to investigate Love waves in functionally graded piezoelectric material (FGPM) layer bonded to a semi-infinite homogeneous solid. The piezoelectric material is polarized in z-axis direction and the material properties change gradually with the thickness of the layer. We here assume that all material properties of the piezoelectric layer have the same exponential function distribution along the x-axis direction. The analytical solutions of dispersion relations are obtained for electrically open or short circuit conditions. The effects of the gradient variation of material constants on the phase velocity, the group velocity, and the coupled electromechanical factor are discussed in detail. The displacement, electric potential, and stress distributions along thickness of the graded layer are calculated and plotted. Numerical examples indicate that appropriate gradient distributing of the material properties make Love waves to propagate along the surface of the piezoelectric layer, or a bigger electromechanical coupling factor can be obtained, which is in favor of acquiring a better performance in surface acoustic wave (SAW) devices. PMID- 17107700 TI - The role of perceptual learning on modality-specific visual attentional effects. AB - Morrone et al. [Morrone, M. C., Denti, V., & Spinelli, D. (2002). Color and luminance contrasts attract independent attention. Current Biology, 12, 1134 1137] reported that the detrimental effect on contrast discrimination thresholds of performing a concomitant task is modality specific: performing a secondary luminance task has no effect on colour contrast thresholds, and vice versa. Here we confirm this result with a novel task involving learning of spatial position, and go on to show that it is not specific to the cardinal colour axes: secondary tasks with red-green stimuli impede performance on a blue-yellow task and vice versa. We further show that the attentional effect can be abolished with continued training over 2-4 training days (2-20 training sessions), and that the effect of learning is transferable to new target positions. Given the finding of transference, we discuss the possibility that V4 is a site of plasticity for both stimulus types, and that the separation is due to a luminance-colour separation within this cortical area. PMID- 17107701 TI - Reactivation and growth of non-culturable indicator bacteria in anaerobically digested biosolids after centrifuge dewatering. AB - Recent literature has reported that high concentrations of indicator bacteria such as fecal coliforms (FCs) were measured in anaerobically digested sludges immediately after dewatering even though low concentrations were measured prior to dewatering. This research hypothesized that the indicator bacteria can enter a non-culturable state during digestion, and are reactivated during centrifuge dewatering. Reactivation is defined as restoration of culturability. To examine this hypothesis, a quantitative polymerase chain reaction (qPCR) method was developed to enumerate Escherichia coli, a member of the FC group, during different phases of digestion and dewatering. For thermophilic digestion, the density of E. coli measured by qPCR could be five orders of magnitude greater than the density measured by standard culturing methods (SCMs), which is indicative of non-culturable bacteria. For mesophilic digestion, qPCR enumerated up to about one order of magnitude more E. coli than the SCMs. After centrifuge dewatering, the non-culturable organisms could be reactivated such that they are enumerated by SCMs, and the conditions in the cake allowed rapid growth of FCs and E. coli during cake storage. PMID- 17107702 TI - Relevance of side reactions in anaerobic reductive dechlorination microcosms amended with different electron donors. AB - In this study we examined the relative importance of side reactions, i.e. the formation of volatile fatty acids (VFA), and the reduction of alternative electron acceptors (nitrate, sulfate, Fe(III)), in enhanced dechlorination microcosms, amended with different electron donors, namely lactate, butyrate, and H(2)+acetate mixture. Dechlorination reactions proceeded at low rates and consequently, nearly all of the reducing equivalents (over 99%) available from electron donor consumption were channeled to (side) reactions other than dechlorination. The relevance of these side reactions was more evident with lactate which was consumed at higher rate than other electron donors. Correspondingly, high levels of VFA and soluble Fe(II) accumulated in the supernatant of lactate-amended microcosms. Ecotoxicity experiments (Lepidium sativum germination tests) also indicated that the supernatant was much more toxic/inhibitory than that of other microcosms. Among the electron donors tested, the H(2)+acetate mixture, yielded the most promising results in terms of extent of dechlorination, negligible accumulation of by-products, and residual groundwater toxicity. Fluorescent In situ Hybridization analysis (FISH) confirmed that H(2)+acetate-amended microcosms were dominated by Dehalococcoides spp., while a higher biodiversity was observed in the cultures fed with lactate or butyrate. Overall, the average amount of donor that was required for the removal of 1micromol of chloride from the contaminant differed greatly among the donors, namely 2.13meq/micromolCl(-) for lactate, 1.01meq/micromolCl(-) for butyrate, and 0.39meq/micromolCl(-) for H(2)+acetate mixture. Interestingly, dechlorinating activity was observed under sulfate-reducing conditions; this suggests that it may not be necessary to deplete the sulfate from the groundwater, for instance by supplying an excess electron donor, in order to achieve substantial dechlorination. Finally, in this study we found that the pathway of anaerobic lactate degradation shifted from the production of acetate and H(2) during active sulfate reduction to acetate and propionate upon sulfate depletion. Energetic considerations that support this finding were presented. PMID- 17107703 TI - Metal bioaccumulation in the greater white-toothed shrew, Crocidura russula, inhabiting an abandoned pyrite mine site. AB - Hepatic and renal concentrations of iron, magnesium, zinc, lead, copper, manganese, mercury, cadmium, molybdenum, chromium, and nickel were quantified in shrews (Crocidura russula) inhabiting a pyrite mine site in Portugal. Several morphological parameters (body weight, residual index, and relative weights) were also examined to clarify the physiological effects of pollution. Shrews from the mine showed increased bioavailability of Fe, Pb, Hg, Cd, Mo, and Ni in comparison with reference specimens. Adult shrews had the highest Cd levels while Cr and Ni concentrations diminished. Intersexual differences were found for Mo and Ni. As a consequence of metal pollution, the relative hepatic weight was higher in shrews from the mine site when compared with reference specimens. These data indicate that C. russula is a good bioindicator of metal pollution. We also evaluated the toxic effects of Pb, Hg, Cd, and Ni, because several shrews from the polluted site showed high concentrations of these metals. To approximate at the real biological impact of abandoned mines, after this first step it is necessary to associate the bioaccumulation levels and morphological effects with other physiological, ecological and genetical biomarkers. PMID- 17107704 TI - Absorption of selenium by Lactuca sativa as affected by carboxymethylcellulose. AB - Several organic compounds of high molecular weight present in soil interact with selenium and may act as active binding agents affecting its availability in soil, and, consequently, selenium uptake by plants. This study is aimed at investigating the effects of polysaccharides on selenium speciation in soil and on selenium absorption by Lactuca sativa L. plants. Three-week-old seedlings were transplanted into pots filled with soil, and sodium selenite at rates of 1.5 and 5mgSekg(-1) of soil, or sodium selenate at a rate of 1.5mgSekg(-1) of soil were applied. Carboxymethylcellulose (CMC) was added to the soil at rates of 0, 3 and 30mgkg(-1) of soil. After 48 and 110d from transplanting plants were harvested, separated into root and shoot, and fresh and dry matter weights were recorded. Total selenium was determined in both soil and plant samples. A sequential extraction was used to investigate the different Se oxidation states and assess the availability of Se in soil after the final harvesting. Both selenite and selenate were absorbed by roots, but plants amended with Se(VI+) showed higher selenium concentration than plants amended with Se(IV+). Selenite appears to be less mobile than selenate both in soil and plants. The addition of carboxymethylcellulose to soil decreased the amount of selenium absorbed by plants. CMC interacted with Se, making it less mobile as evidenced by the increase in the insoluble fractions. The insoluble Se forms in soil may represent environmental Se sinks potentially available for plants if the substrate is re used for subsequent growth cycles and selenium species are mobilized as a result of biological and chemical processes. PMID- 17107705 TI - Ex vivo microvesicle formation after prolonged ischemia in renal transplantation. AB - INTRODUCTION: Patients with chronic renal failure suffer from dysfunction in coagulation. Kidney transplantation induces inflammatory reactions and thus activation of platelets. Activated platelets, in turn, form microvesicles by shedding. These microvesicles have been shown to have coagulant activities. Activated platelets in prolonged cold ischemia were associated with delayed graft function and inferior survival. We investigated ex vivo formation of microvesicles in kidney transplantation and the influence of cold graft storage on microvesicles. METHODS: 20 patients (47.4+/-10.6 years (mean+/-SD)) undergoing transplantation were included in the study after written informed consent. Dependent on cold preservation time of transplanted kidneys, recipients were allocated into two groups with 10.4+/-6.1 h (group 1) and 23.7+/-3.8 h (group 2) preservation time, respectively. Blood samples were drawn before anesthesia, 12 h, 2, 7 and 14 days after transplantation. To evaluate microvesicle release, samples were activated with thrombin-receptor-activating-peptide-6 (TRAP) or adenosine-di-phosphate. Microvesicles were counted as percentage of platelets smaller than a predetermined size in flow cytometry. RESULTS: Platelet derived ex vivo microvesicle formation was significantly higher up to 48 h after transplantation when stimulated with TRAP in group 1. Platelet count was significantly higher compared to baseline values in the short-term ischemia group but not with long-term ischemia. Creatinine was significantly lower at study end compared to baseline with no differences between both groups. CONCLUSIONS: Lower platelet microvesicle formation after ex vivo stimulation with TRAP was associated with longer graft ischemia time. This may be a sign of former activation of platelets which could influence graft function and survival. PMID- 17107706 TI - How gene-stress-behavior interactions can promote adolescent alcohol use: the roles of predrinking allostatic load and childhood behavior disorders. AB - A variety of environmental and genetic factors modulating the risk for alcoholism have been described, which predominantly act by interacting with each other. For example, the family, peers and society determine the level of exposure to stress and alcohol, while genes modulate how sensitive an individual responds to both. The resulting behaviors feed back to the social environment, modulating and in the worst case increasing further stress exposure. We here review neurobiological evidence how such a process of mutual interaction can involve and affect drinking. In at-risk adolescents it may have been in force for many years before they have their first alcoholic drink, increasing their risk for addiction by generating allostatic load. As an example, psychiatric disorders involving attention deficit, hyperactivity, or disruptive behaviors first evolve during childhood and are influenced by all the above factors. They are also strongly associated with harmful adolescent drinking and later alcohol use disorders. One important implication of this concept is that issues such as family adversity, adolescent psychiatric disorders, or adolescent drinking might not only be associated with, but causally related to, the risk for later addiction. They are targets for preventive interventions, which should start as early as possible in subjects at-risk. PMID- 17107707 TI - The effects of acute and chronic administration of corticosterone on rat behavior in two models of fear responses, plasma corticosterone concentration, and c-Fos expression in the brain structures. AB - The aim of this paper was to examine changes in rat emotional behavior, and to find the brain structures, which are involved in the mediation of behavioral effects, related to the repeated administration of glucocorticoids. The effects of acute and chronic pretreatment of rats with two doses of corticosterone (5 and 20 mg/kg) were analyzed in two models of fear responses: neophobia-like behavior in the open field test, and freezing reaction in the conditioned fear test. Behavioral effects of repeated glucocorticoid administration were compared to changes in blood total corticosterone concentration, and expression of immediate early gene (c-Fos) in brain structures. It was found that acute administration of corticosterone (90 min before tests) enhanced rat exploratory behavior, and decreased freezing reaction. On the other hand, repeated administration of corticosterone (for 25 days, the final injection 90 min before contextual fear conditioning training) decreased plasma corticosterone concentration, inhibited exploratory behavior, enhanced freezing responses on retest and produced a complex pattern of changes in c-Fos expression, stimulated by exposure of rats to the aversively conditioned context. Aversive context induced c-Fos in the magnocellular neurons of the hypothalamic paraventricular nucleus (mPVN), dentate gyrus (DG), cingulate cortex area 1 (Cg1), and primary motor cortex (M1). In rats chronically treated with corticosterone this effect was attenuated in the mPVN and DG, enhanced in the M1, and additionally observed in the CA1, CA2 layers of the hippocampus, and in the central nucleus of amygdala (CeA), in comparison to control animals not subjected to contextual fear test. In sum, the present data suggest that chronic corticosterone treatment enhances the activity of primary motor cortex and CeA with subsequent improvement of memory of aversive events, and simultaneously stimulates a negative feedback mechanism operating in PVN with ensuing decrease in blood corticosterone concentration. PMID- 17107708 TI - Enzyme-degradable phosphorylcholine porous hydrogels cross-linked with polyphosphoesters for cell matrices. AB - Biodegradable highly porous hydrogels composed of poly [2-methacryloyloxyethyl phosphorylcholine (MPC)] cross-linked with polyphosphoesters have been prepared as novel cellular matrices. Well-controlled porous hydrogels were fabricated by using potassium hydrogen carbonate as a porogen salt for forming gas. This process enabled the homogeneous expansion of pores within the polymer hydrogel matrices, leading to well-interconnected high porosity. The mechanical properties of the hydrogels were influenced by the cross-linking density and porous structure. Hydrolysis and enzymatic digestion of the hydrogels were determined under basic conditions. The cross-linking density and porosity influenced the rate of degradation of the hydrogels. Acceleration of the degradation with alkaline phosphatase was also observed. Cultivation of mouse osteoblastic cell (MC3T3-E1) was performed in the highly porous hydrogels and cell viability was well maintained. The rate of cell proliferation also was relatively increased with an increase in the amount of polyphosphoesters in the hydrogel. Basic fibroblast growth factor (bFGF) was physically absorbed by the hydrogels and effectively induced cell proliferation. In conclusion, the porous hydrogels prepared in this study contributed a suitable environment for three-dimensional cell cultivation and may be useful for cell and tissue matrices. PMID- 17107709 TI - Peroxidation of polyunsaturated phosphatidyl-choline lipids during electroformation. AB - Giant unilamellar vesicles (GUVs) have been utilized both as model systems to study the physico-chemical properties of biomembranes and as host materials for investigating biological processes in microbioreactors. GUVs are commonly formed by an electroformation technique. However, there is a concern that the electric fields applied during electroformation can peroxidize lipid acyl chains, thereby altering the phospholipid composition and material properties of the synthesized vesicles. Here in this paper, we report the effect of electroformation on the extent of peroxidation of a number of polyunsaturated phosphatidyl-choline lipids (PULs). Specifically, we detected peroxidation byproducts (malonaldehydes and conjugated dienes) of the following lipids utilizing UV/Vis spectroscopy: dilinoleoyl phosphatidyl-choline (DLPC) (di-18:2 PC), dilinolenoyl phosphatidyl choline (DNPC) (di-18:3 PC), diarachidonoyl phosphatidyl-choline (DAPC) (di-20:4 PC), and didocosaheexaenoyl phosphatidyl-choline (DHA) (di-22:6 PC). The results indicate that PC PULs lipids are prone to peroxidation, with increasing unsaturation levels leading to higher levels of peroxidation byproducts. The levels of peroxidation byproducts of DAPC were found to depend linearly on the strength of the electric field, indicating that the observed effects were due to the applied electric field. Lipid peroxidation can affect a number of important membrane properties, including domain formation and mechanical stability. Thus, alteration of the chemical composition of polyunsaturated lipids (PULs) by the electroformation technique can potentially complicate the interpretation of experimental studies that utilize GUVs composed of PULs. PMID- 17107710 TI - Oxytocin selectively increases ERalpha mRNA in the neonatal hypothalamus and hippocampus of female prairie voles. AB - During neonatal development exogenous oxytocin increases ERalpha immunoreactivity in the hypothalamus of female prairie voles. The purpose of this study was to determine if the increase in ERalpha is associated with an increase in ERalpha mRNA expression and to determine if the effect is specific to ER subtype or if oxytocin also influences ERbeta mRNA expression. On the day of birth female prairie vole pups were treated with oxytocin, an oxytocin antagonist, or saline. Brains were collected and RT-PCR was used to determine the effect of treatment on ER mRNA production in the hypothalamus, hippocampus, and cortex. Within 2h of treatment oxytocin significantly increased ERalpha mRNA expression in the hypothalamus and hippocampus, but not the cortex, while inhibiting the effects of endogenous oxytocin reduced the expression of ERalpha mRNA in the hippocampus. Neonatal treatment did not affect the expression of ERbetamRNA. The results demonstrate that the effects of oxytocin treatment are region and ER subtype specific and that during the neonatal period oxytocin can affect the expression of ERalpha by altering message production. The regional specific changes in ERalpha mRNA expression in females are consistent with studies examining the behavioral and physiological effects of neonatal manipulation of oxytocin in females. PMID- 17107711 TI - The role of neuropeptide FF (NPFF) in the expression of sensitization to hyperlocomotor effect of morphine and ethanol. AB - Neuropeptide FF (NPFF) has been characterized as an endogenous anti-opioid peptide because its intraventricular injection (icv) reversed morphine- and stress-induced analgesia, and precipitates withdrawal syndrome in morphine dependent rats. The role of NPFF in other aspects of drug dependence is unknown. Therefore, the aim of this study was to determine NPFF influence on the expression of sensitization to the morphine-induced hyperlocomotion. As the opioid system plays a role in ethanol effects, the influence of NPFF on the expression of sensitization to hyperlocomotor effect of ethanol was also investigated. Our study indicated that acute administration of NPFF (5, 10, 20nmol, icv) inhibited the expression of morphine-induced sensitization at doses of 10 (P<0.05) and 20nmol (P<0.01), and also inhibited ethanol-induced sensitization at a dose of 20nmol (P<0.01). Furthermore, NPFF inhibited the acute locomotor effect of morphine (10 and 20nmol) but not that of ethanol. NPFF, given alone, did not change the locomotor activity of mice and did not disturb motor coordination of animals in the rotarod test. In conclusion, our experiments indicated that NPFF attenuated the acute morphine locomotion and the expression of sensitization to locomotion. We anticipate that NPFF may be involved in both of these effects. PMID- 17107712 TI - Complement expression in common carp (Cyprinus carpio L.) during infection with Ichthyophthirius multifiliis. AB - A real-time PCR assay for determination of the complement response to infection with the ectoparasite Ichthyophthirius multifiliis in carp is presented. Specific primers were designed for selected genes representing the three pathways of the carp complement system. The investigated complement molecules were C1r/s, C3, C4, C5, factor I, factor B/C2-A (Bf/C2-A), mannose-binding lectin (MBL) and MBL associated serine protease (MASP). The expression of the selected genes was analyzed on RNA extracts from skin, liver, and whole blood from carp at 3, 12, 24, 36, and 48 h post-infection (pi) with I. multifiliis. A pronounced up regulation of Bf/C2-A, in skin, blood, and liver (250-, 60-, and 4-fold respectively), was observed at later sampling points pi (24-48 h). In addition, an intermediate (from 5 to 13-fold) down-regulation of MASP was observed in skin and liver samples at 36 and 48 h pi with respect to control fish. MBL was expressed only in liver and no variation in the transcription level of this lectin was observed. Complement factor C3 was significantly up-regulated in liver (4-fold up-regulation, 24 h pi). The presented results indicate that infection with the parasite I. multifiliis in carp to a large extent stimulates the expression of complement molecules. Moreover, the dramatic and early up regulation of Bf/C2-A in skin indicates a role of this molecule as an acute-phase reactant. Furthermore, our study confirms the role of fish skin as an important extra-hepatic site of expression of complement molecules as well as an active regulator of complement expression. Expression of some of the components of the complement system in blood suggests that leukocytes in carp act as an important extra-hepatic source of complement molecules. PMID- 17107713 TI - Trends in solventless sample preparation techniques for environmental analysis. AB - The paper presents recent trends in solventless sample preparation techniques for environmental analysis. First, a general classification of solventless methods is given. Next, three of them, treated as preferable techniques, i.e. SPME, SDME and HS, are presented in detail, with respect to their usability and effectiveness for environmental samples. Examples of all discussed techniques are given in the tables. PMID- 17107714 TI - Matrix solid phase dispersion (MSPD). AB - A review of the many uses of matrix solid phase dispersion (MSPD) in the extraction and analysis of a variety of compounds from a range of samples is provided. Matrix solid phase dispersion (MSPD) has found particular application as a somewhat generic analytical process for the preparation, extraction and fractionation of solid, semi-solid and/or highly viscous biological samples. Its simplicity and flexibility contribute to it being chosen over more classical methods for these purposes. MSPD is based on several simple principles of chemistry and physics, involving forces applied to the sample by mechanical blending to produce complete sample disruption and the interactions of the sample matrix with a solid support bonded-phase (SPE) or the surface chemistry of other solid support materials. These principles are discussed as are the factors to be considered in conducting a MSPD extraction. PMID- 17107715 TI - Application of molecularly imprinted polymers to solid-phase extraction of analytes from real samples. AB - A review is presented of recent developments in the use of molecularly imprinted polymers (MIPs) as selective materials for solid-phase extraction. Compared with traditional sorbents, MIPs can not only concentrate but also selectively separate the target analytes from real samples, which is crucial for the quantitatively determination of analytes in complex samples. Consequently, as one of the most effective sorbents, MIPs have been successfully applied to the pretreatment of analytes in foods, drugs, and biological and environmental samples in the past five years. PMID- 17107716 TI - Dissociation of cognitive from affective components of theory of mind in schizophrenia. AB - Patients suffering from schizophrenia show impaired emotional and social behavior, such as misinterpretation of social situations and lack of theory of mind. However, there is conflicting evidence regarding their ability to perform on theory of mind tasks. Based on previous findings with patients suffering from prefrontal damage, the present study suggests that the behavioral deficit of schizophrenic patients may be due to impaired 'affective theory of mind' abilities, rather than to a general impairment in theory of mind. To test this hypothesis we assessed the ability of 22 schizophrenic patients and 55 age matched healthy controls, to judge first and second order affective vs. cognitive mental state attribution, based on eye gaze. The relationships between negative and positive symptoms of schizophrenia, and affective and cognitive theory of mind were also assessed. Results indicated that while healthy controls made fewer errors on affective as compared to cognitive theory of mind conditions, schizophrenic patients showed a less prominent trend. Although the pattern of reaction time did not differ significantly between groups, the patients made significantly more errors in the affective conditions, as compared to controls. Furthermore, correlation analysis indicated that impaired affective theory of mind in these patients correlated with their level of negative symptoms. These results indicate that individuals with high level of negative symptoms of schizophrenia may demonstrate selective impairment in their ability to attribute affective mental states. These findings offer new insight into the affective facets of social behavior that may underlie the profound behavioral disturbances observed in schizophrenia. PMID- 17107717 TI - TNF-alpha is critical for ischemia-induced leukostasis, but not retinal neovascularization nor VEGF-induced leakage. AB - Vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF alpha) show significant overlap with regard to their effects in the eye. It has been postulated that VEGF-induced leukostasis, breakdown of the blood-retinal barrier, and ischemia-induced retinal neovascularization may be mediated, at least in part, through TNF-alpha. In this study, we used mice deficient in TNF alpha to test our hypothesis. Compared to wild type mice, TNF-alpha-deficient mice showed an 80% reduction in leukocyte accumulation in retinal vessels after intravitreous injection of VEGF, and 100% reductions after intravitreous injections of interleukin-1beta (IL-1beta) or platelet-activating factor (PAF). The increase in retinal vascular permeability induced by injection of PAF was significantly reduced in mice lacking TNF-alpha, but VEGF- and IL-1beta-induced leakage was unaffected. Compared to wild type mice with oxygen-induced ischemic retinopathy, TNF-alpha-deficient mice with ischemic retinopathy showed significantly reduced leukostasis and mild reduction in vascular leakage, but no significant difference in retinal neovascularization. These data suggest that TNF alpha mediates VEGF-, IL-1beta-, and PAF-induced leukostasis and vascular leakage mediated by PAF, but not leakage caused by VEGF or IL-1beta. Ischemia-induced retinal neovascularization, which has previously been shown to require VEGF, does not require TNF-alpha and is unaffected by attenuation of leukostasis. PMID- 17107718 TI - Metabolism of glucagon-like peptide-2 in pigs: role of dipeptidyl peptidase IV. AB - Little is known about the metabolism of the intestinotropic factor glucagon-like peptide-2 (GLP-2); except that it is a substrate for dipeptidyl peptidase IV (DPP IV) and that it appears to be eliminated by the kidneys. We, therefore, investigated GLP-2 metabolism in six multicatheterized pigs receiving intravenous GLP-2 infusions (2 pmol/kg/min) before and after administration of valine pyrrolidide (300 mumol/kg; a well characterized DPP-IV inhibitor). Plasma samples were analyzed by radioimmunoassays allowing determination of intact, biologically active GLP-2 and the DPP-IV metabolite GLP-2 (3-33). During infusion of GLP-2 alone, 30.9+/-1.7% of the infused peptide was degraded to GLP-2 (3-33). After valine-pyrrolidide, there was no significant formation of the metabolite. Significant extraction of intact GLP-2 was observed across the kidneys, the extremities (represented by a leg), and the splanchnic bed, resulting in a metabolic clearance rate (MCR) of 6.80+/-0.47 ml/kg/min and a plasma half-life of 6.8+/-0.8 min. Hepatic extraction was not detected. Valine-pyrrolidide addition did not affect extraction ratios significantly, but decreased (p=0.003) MCR to 4.18+/-0.27 ml/kg/min and increased (p=0.052) plasma half-life to 9.9+/-0.8 min. The metabolite was eliminated with a half-life of 22.1+/-2.6 min and a clearance of 2.07+/-0.11 ml/kg/min. In conclusion, intact GLP-2 is eliminated in the peripheral tissues, the splanchnic bed and the kidneys, but not in the liver, by mechanisms unrelated to DPP-IV. However, DPP-IV is involved in the overall GLP-2 metabolism and seems to be the sole enzyme responsible for N-terminal degradation of GLP-2. PMID- 17107719 TI - Anomalies of the coronary arteries originating from the right sinus of Valsalva. (1) Single coronary artery originating from the right sinus associated with fusion of the left and the non coronary cusp and atrophy of the left coronary ostium (2)Three separate coronary arteries originating from the right sinus of Valsalva. AB - We report three patients with the entire coronary origin arising from the right sinus of Valsalva. The first patient had a single right coronary ostium associated with a bi-leaflet aortic valve. The second patient was admitted with ST-elevation myocardial infarction (STEMI) for primary PCI. The third patient had 3 isolated ostia, all originating from the right sinus of Valsalva. Coronary anomalies are associated with increased mortality, depending on the myocardium at risk. A left main originating from the right coronary sinus is supplying a greater extent of the myocardium and is associated both with an increased incidence of symptoms and of sudden cardiac death. The possibility of such an artery anomaly should always be considered in young individuals with a history of chest pain or syncope. PMID- 17107720 TI - Increased risk of cardiac death in primary hyperparathyroidism: what is a role of electrical instability? AB - BACKGROUND: Increased risk of death in patients with primary hyperparathyroidism (PHPT) is referred to cardiovascular complications induced by hypercalcemia. At the moment the role of bioelectrical risk and of enhanced sympathetic activity, not related to cardiovascular complications, is unknown in PHPT patients. METHODS: The study was designed to evaluate the QT parameters and sympathovagal balance in 28 PHPT uncomplicated patients compared to 29 healthy subjects (HS). Nine of 28 patients were restudied after parathyroidectomy. Standard ECG, short and 24-h ECG monitoring were performed to calculate QT parameters and Heart Rate Variability (HRV). RESULTS: QTc interval resulted shorter in PHPT patients than in HS and QTc dispersion resulted higher in PHPT patients than HS. The physiological adaptation of QT length to R-R interval was lacking in PHPT patients. Enhanced sympathetic tone was found in PTHP patients than controls. These data were confirmed after parathyroidectomy. CONCLUSION: Our findings revealed an increased risk to develop life-threatening arrhythmias in PHPT patients due to bioelectrical instability induced by hypercalcemia. Parathyroidectomy doesn't seem to reverse this abnormality. PMID- 17107721 TI - Emerging trends of community acquired infective endocarditis. AB - BACKGROUND: To obtain epidemiological information and to evaluate the emerging trend of incidence and clinical features of community acquired infective endocarditis (IE) in Hong Kong. METHODS: Population-based survey in a regional hospital in Hong Kong which served 0.55 million population over an 11-year period was conducted. 172 patients diagnosed to have IE between 1995 and 2005 were analyzed. Incidence, underlying heart disease, causative microorganisms and clinical outcomes of IE were studied. RESULTS: The standardized annual incidence of community acquired IE was 2.8 cases per 100,000 person-year and remain stable over the past 10 years in Hong Kong (P=0.57 for trends). The most common underlying heart diseases for IE were intravenous drug addict (30%), followed by chronic rheumatic heart disease (18%). There was a time trend of increasing age in patients suffered from IE, both in patients with or without intravenous drug abuse (overall P=0.004). Although there were no significant changes in the overall proportion of IE patients with different underlying etiologies, site of IE involvement and types of microorganisms, non-addict patients showed an increasing trend of staphylococcal IE during the study period (P=0.01). The adverse clinical outcome of IE during the study period remained unchanged (P=0.71). CONCLUSIONS: The results of this study have demonstrated that IE continued to be an important disease with stable incidence and high morbidity and mortality over time in Asia. Furthermore, our study also highlighted that IE is an emerging disease with continuously changing clinical and microbiologic patterns, and significant differences are still present between different countries. PMID- 17107722 TI - Left main coronary artery stenosis associated with extremely long fusiform aneurysm. AB - Coronary artery aneurysm is defined as coronary dilatation, which exceeds the diameter of normal adjacent segment or the diameter of the patient's largest coronary vessel by 1.5 times [Syed M, Lesch M: Coronary artery aneurysms: a review. Prog Cardiovasc Dis 1997; 40:77-10 84 [1]]. Most studies show an incidence of 1% to 2% [Barettella MB, Bott-Silverman C. Coronary artery aneurysm: an unusual case report and a review of the literature. Cathet Cardiovasc Diagn 1993; 29:57-61 [2]]. Left main coronary aneurysms (LMCA) are even more rare; in a study by involving 22,000 coronary angiograms an occurrence rate of 0.1% has been found [Topaz O, DiSciascio G, Cowley MJ, Goudreau E, Soffer A, Nath A et al. Angiographic features of left main coronary artery aneurysms. Am J Cardiol 1991; 67:1139-1142 [3]]. The majority of patients present with ischemic symptoms secondary to the coexisting atherosclerotic lesions and most of the coronary aneurysms are incidentally diagnosed by coronary angiography. This report details the exceptional case of a 62-year-old patient with "unusually long fusiform" aneurysm of the left main coronary artery associated with critical left main coronary artery distal stenosis involving the ostia of left anterior descending and left circumflex coronary artery. With this case the possible anatomic determinants of the type, location, and mechanism of aneurysm formation were also discussed. PMID- 17107723 TI - Two- and three-dimensional echocardiographic imaging of massive left intraventricular thrombosis complicating fatal heart failure. AB - These images describe a case of massive left ventricular (LV) thrombosis complicating fatal heart failure in a 61 year-old patient with history of myocardial infarction, apical aneurysm, and no previous evidence of LV thrombosis. Two-dimensional echocardiography showed that the thrombus obliterated 34% of LV volume at end-diastole. Three-dimensional echocardiography allowed higher definition of thrombus architecture and dimensions, revealing obliteration of 40% of LV volume at end-diastole. Careful assessment of thromboembolic risk to guide therapeutic decision making should be considered mandatory in subjects with a large extent of wall asynergy, even in those with no previous evidence of LV thrombosis. Three-dimensional echocardiography may be considered an accurate technique for the characterization of LV thrombi in these patients. PMID- 17107724 TI - Influence of epicardial adipose tissue and adipocytokine levels on cardiac abnormalities in visceral obesity. AB - Adipocytokine levels and visceral adipose tissue (VAT) seem to be associated with some cardiac abnormalities and a role of visceral fat in predisposing to cardiac dysfunction, possibly through a low-grade state of inflammation, has been demonstrated. In this study we firstly show that elevated levels of both monocyte chemoattractant protein 1 (MCP-1) and soluble IL-6 receptor/interleukin-6 (sIL 6R/IL-6) complex are closely correlated with epicardial fat thickness. PMID- 17107725 TI - Atresia of the coronary sinus ostium: surgical implications. AB - We report a neonatal case of atresia of the coronary sinus ostium draining into a persistent left superior caval vein, associated with aortic arch hypoplasia, ventricular septal defect and mitro-aortic discontinuity. In this setting, left superior caval vein receives the coronary flow, which is directed into the innominate vein and the right atrium. It is mandatory to perform a precise anatomical definition of this unusual cardiac malformation, before repair of associated cardiac defects. Indeed, the interruption of coronary venous drainage could potentially lead to myocardial ischemia and necrosis. PMID- 17107726 TI - Impact of hospital and physician characteristics on medical expenditures for acute myocardial infarction hospitalization. AB - This study aims to examine the impact of length of stay, hospital characteristics, physician characteristics and other factors on the expenditures of hospitalization for acute myocardial infarction (AMI) under Taiwan's National Health Insurance program. This study uses data collected from the Taiwan's National Health Research Institute's 2001-2003 National Health Insurance Research Database. We estimated contributors to increased expenditures of hospitalization using three-stage least square regression model. The hospital expenditures for the treatment of AMI averaged NT$126,366 (US$3829, US$1=NT$33) per discharge, with the largest proportion (27%) spent on room expenditures. They were strongly impacted by length of stay, increasing around 4.8% per day. We conclude that hospital expenditures for the treatment of AMI patients may vary widely depending on the characteristics of the hospital and physicians that provide them care. PMID- 17107727 TI - Late thrombosis of a drug-eluting stent overlapped in the in-stent restenotic lesion of a bare metal stent. AB - Late stent thrombosis occurred in the lesion of a sirolimus-eluting stent implanted 6 months previously for an in-stent restenosis lesion in the distal right coronary artery. Seventeen days before admission due to acute myocardial infarction this time, aspirin was discontinued for colon polypectomy. Ticlopidine had been discontinued 3 months before the discontinuation of aspirin. In drug eluting stent era, the interventional strategy and antiplatelet therapy require long term attention. PMID- 17107728 TI - Gone but not forgotten: the effects of cancelled intentions on the neural correlates of prospective memory. AB - Event-related brain potentials (ERPs) were used to examine the neural correlates of the prospective interference effect and the effects of canceling an intention on the neural correlates of prospective memory. The response time data revealed a prospective interference effect that was associated with the engagement of prospective retrieval mode and item checking. The ERP data revealed that item checking was associated with sustained activity over the frontal and occipital parietal regions of the scalp beginning at around 300 ms after stimulus onset. The ERP data also revealed that canceling an intention may have blocked the retrieval of the intention from memory when the prospective cue was encountered and led to a significant attenuation in the degree that the neural correlates of cue detection and post-retrieval processes were expressed. PMID- 17107729 TI - Synergistic effect of chitosan and Cryptococcus laurentii on inhibition of Penicillium expansum infections. AB - This study was conducted to determine the efficacy of chitosan at different concentrations with various intrinsic viscosities alone, and in its combination with a yeast antagonist Cryptococcus laurentii in reducing the blue mold rot caused by Penicillium expansum in apple fruit. The results indicated that application of chitosan alone was effective in inhibiting the blue mold rot in apple fruit wounds, especially with the high concentrations and low viscosities. But its efficacy was declining with the incubation time so that chitosan alone could not provide enduring protection of apple fruit from P. expansum infections. When applied at the concentration range from 0.001 to 0.1% (wt/vol), chitosan did not influence the population growth of C. laurentii in vivo, whereas it markedly repressed the yeast growth as its concentrations were increased up to 0.25% (wt/vol) or higher. Moreover, combination of chitosan and C. laurentii resulted in a synergistic inhibition of the blue mold rot, being the most effective at the optimal concentration of 0.1% of chitosan with the lowest viscosity (12 cP). The possible mode of action of the combination of chitosan and C. laurentii was discussed. PMID- 17107730 TI - High levels of ochratoxin A in licorice and derived products. AB - The ochratoxin A (OTA) content of 30 samples of licorice root and derived products (licorice-confectionery, licorice block, and licorice extract) was analyzed by a standard HPLC-fluorescence technique and confirmed by methyl-ester formation. All analyzed samples of licorice and derived products were found to contain ochratoxin A, and some of them showed extremely high concentrations up to 252.8 ng/g of OTA. Highest levels of ochratoxin A were found in dry licorice root, averaging 63.6 ng/g, while mean contents in fresh licorice root were 9.2 ng/g. Licorice-confectionery (sweets) contained 3.8 ng/g of OTA. Ochratoxin A was also abundant in two licorice derivatives, liquid licorice extract (16.0 ng/g) and solid licorice block (39.5 ng/g). The ochratoxin levels found in licorice and derived products are higher than those reported in the literature for other food commodities. The experiments of OTA transfer into the tea beverages showed that almost 5% of the OTA present in dry licorice root is transferred to the corresponding decoction tea, whereas only 1% of OTA remains in infusion tea. The significance of the levels of ochratoxin A in licorice and its derivatives is discussed in the context of existing data on ochratoxin contamination in foods. PMID- 17107732 TI - Reefer madness? Assessing the effects of cannabinoids with a less jaundiced eye. PMID- 17107731 TI - Rosiglitazone attenuates suppression of RXRalpha-dependent gene expression in inflamed liver. AB - BACKGROUND/AIMS: A recently determined target of lipopolysaccharide (LPS) and cytokine signaling in liver is the central Type II nuclear receptor (NR) heterodimer partner, retinoid X receptor alpha (RXRalpha). We sought to determine if Rosiglitazone (Rosi), a peroxisome proliferator activated receptor gamma (PPARgamma) agonist with anti-inflammatory properties, can attenuate LPS and cytokine-induced molecular suppression of RXRalpha-regulated genes. METHODS: In vivo, mice were gavage-fed Rosi for 3 days, prior to intraperitoneal injection of LPS, followed by harvest of liver and serum. In vitro, HepG2 cells were treated with IL-1beta, +/- short-term Rosi pretreatment. RNA was analyzed by quantitative RT-PCR, while nuclear and cytoplasmic proteins were analyzed by immunoblotting and gel shifts. RESULTS: Rosi attenuated LPS-mediated suppression of RNA levels of several Type II NR-regulated genes, including bile acid transporters and the major drug metabolizing enzyme, Cyp3a11, without affecting cytokine expression, suggesting a novel, direct anti-inflammatory effect in hepatocytes. Rosi suppressed the inflammation-induced nuclear export of RXRalpha, in both LPS injected mice and IL-1beta-treated HepG2 cells, leading to maintenance of nuclear RXRalpha levels and heterodimer binding activity. CONCLUSIONS: Rosi directly attenuates the suppressive effects of inflammation-induced cell signaling on nuclear RXRalpha levels in liver. PMID- 17107733 TI - Intracellular accumulation of hepatitis C virus proteins in a human hepatoma cell line. AB - BACKGROUND/AIMS: The establishment of HCV replicon systems strongly improved the research on the replication processes but poorly advanced our knowledge on the subcellular localization of the structural glycoproteins, mainly due to their low expression. We sought to verify whether reinforcing E1E2 expression in the context of both HCV genomic and subgenomic replicon from either homologous or heterologous strains leads to formation of supramolecular structures including structural and nonstructural proteins. METHODS: Robust expression of HCV glycoproteins was achieved by stable expression of E1E2p7 from genotype 1a and 1b. RESULTS: In these cells, E1 and E2 triggered the formation of dot-like structures in which they co-localized with core and the nonstructural proteins NS3 and NS5A. Confocal microscopy analyses suggested that accumulation of HCV proteins occurs in an ER-derived subcompartment. Moreover, by labeling de novo synthesized HCV RNA, we showed that these structures constitute a site of viral RNA synthesis. CONCLUSIONS: Expression in trans of HCV glycoproteins in the context of replicative viral genome or subgenome generates accumulation of structural and nonstructural viral proteins in peculiar cytoplasmic structures. The simultaneous presence of viral RNA, structural and nonstructural protein suggests that these complexes represent not only sites of HCV replication but also potential places of viral pre-budding. PMID- 17107734 TI - Interferon therapy in HBeAg positive chronic hepatitis reduces progression to cirrhosis and hepatocellular carcinoma. AB - BACKGROUND/AIMS: The long-term outcomes of interferon-alpha (IFN-alpha) therapy in hepatitis B e antigen (HBeAg) seropositive patients remain controversial. This study was conducted to address this issue. METHODS: The long-term outcomes were compared in 233 IFN-treated patients and 233 well-matched untreated controls. RESULTS: The cumulative incidence at the end of 15 years of follow-up (median 6.8 years, range 1.1-16.5 years) in the IFN-treated patients and controls was: HBeAg seroconversion 74.6% vs. 51.7% (P=0.031); hepatitis B surface antigen (HBsAg) seroclearance 3% vs. 0.4% (P=0.03); cirrhosis 17.8% vs. 33.7% (P=0.041); and hepatocellular carcinoma (HCC) 2.7% vs. 12.5% (P=0.011). Significant reduction of HCC was only observed in patients with pre-existing cirrhosis (P<0.01). Compared with untreated controls with persistent HBeAg, HBeAg seroconverters in untreated and IFN-treated group showed significantly lower incidence of cirrhosis and HCC (P=0.003-0.031), while non-seroconverters of IFN-treated group had marginally significant lower incidence of cirrhosis (P=0.065). Multivariate analysis showed that IFN therapy, HBeAg seroconversion and genotype B HBV infection are independent factors for better long-term outcomes. CONCLUSIONS: IFN therapy reduces cirrhosis and HCC development. PMID- 17107735 TI - Long-term portosystemic shunt patency as a determinant of outcome in Budd-Chiari syndrome. AB - BACKGROUND/AIMS: Portosystemic shunting, whether surgical or transjugular intrahepatic, has been a cornerstone of therapy for Budd-Chiari syndrome. However, the long-term impact of shunt dysfunction remains unknown. We have assessed this long-term impact in patients with surgical shunting. METHODS: Thirty-nine consecutive patients operated on between 1978 and 2000 were analyzed using time-dependent multivariate Cox model. RESULTS: Median follow-up was 110 months. Prosthetic shunts and high preshunt portal venous pressure were predictors of subsequent shunt dysfunction. Among 19 patients with persistently patent shunt, as compared to 20 patients with shunt dysfunction, 1 versus 18 developed refractory ascites; 1 versus 7 had variceal bleeding; 7 versus 2 had encephalopathy; 3 versus 11 (55%) died or underwent liver transplantation; and 0 versus 10 died from end-stage liver disease. Shunt dysfunction was associated with a shorter survival (p=0.001). Out of 20 patients with shunt dysfunction, seven had successful revision of the shunt. None of these seven patients had refractory ascites after revision or died from end-stage liver disease. CONCLUSIONS: In patients with Budd-Chiari syndrome treated with portosystemic shunting, shunt dysfunction has a major impact on morbidity and mortality. PMID- 17107736 TI - Tolerability of Peg interferon-alpha2b and Ribavirin therapy in patients with chronic hepatitis C and glucose-6-phosphate dehydrogenase deficiency. PMID- 17107737 TI - Intraocular implants for extended drug delivery: therapeutic applications. AB - An overview of ocular implants with therapeutic application potentials is provided. Various types of implants can be used as slow release devices delivering locally the needed drug for an extended period of time. Thus, multiple periocular or intraocular injections of the drug can be circumvented and secondary complications minimized. The various compositions of polymers fulfilling specific delivery goals are described. Several of these implants are undergoing clinical trials while a few are already commercialized. Despite the paramount progress in design, safety and efficacy, the place of these implants in our clinical therapeutic arsenal remains limited. Miniaturization of the implants allowing for their direct injection without the need for a complicated surgery is a necessary development avenue. Particulate systems which can be engineered to target specifically certain cells or tissues are another promising alternative. For ocular diseases affecting the choroid and outer retina, transscleral or intrasscleral implants are gaining momentum. PMID- 17107738 TI - Brain cancer diagnosis and therapy with nanoplatforms. AB - Treatment of brain cancer remains a challenge despite recent improvements in surgery and multimodal adjuvant therapy. Drug therapies of brain cancer have been particularly inefficient, due to the blood-brain barrier and the non-specificity of the potentially toxic drugs. The nanoparticle has emerged as a potential vector for brain delivery, able to overcome the problems of current strategies. Moreover, multi-functionality can be engineered into a single nanoplatform so that it can provide tumor-specific detection, treatment, and follow-up monitoring. Such multitasking is not possible with conventional technologies. This review describes recent advances in nanoparticle-based detection and therapy of brain cancer. The advantages of nanoparticle-based delivery and the types of nanoparticle systems under investigation are described, as well as their applications. PMID- 17107739 TI - Expansion of HIV/AIDS in China: lessons from Yunnan Province. AB - In this article we systematically and critically review the Chinese and English language literature on human immunodeficiency virus (HIV)-related studies in Yunnan Province, Southwestern China. Yunnan Province had the first Chinese HIV outbreak and is still the worst affected area in the nation. Since 1989, HIV infection has extended from injecting drug users into the general population through sexual transmission. Since the economic reform of the 1980s, changed social norms and increased migration have spawned increases in HIV-related risk behaviors such as drug use and commercial sex work. A smaller size of "bridge" populations and lower sexual contact rates between persons in "bridge" and general populations may explain the slower expansion of the HIV epidemic in Yunnan compared to nearby Southeast Asian nations. In 2004, women in antenatal care had a 0.38% HIV prevalence province wide, although >1% infection rates are seen in those counties with high injection drug rates. Patterns of drug trafficking have spread the unusual recombinant HIV subtypes first seen in Yunnan to far-flung regions of China. Increased efforts of Yunnan's HIV control program are correlated with an improved general HIV awareness, but risk behaviors continue at worrisome rates. Future efforts should focus on changing risk behaviors, including harm reduction and condom promotion, especially among the "bridge" groups. The resurgence of commercial sex work in Yunnan, and the high frequency of workers migrating into provinces far from home and family are all sociocultural factors of considerable importance for future HIV and sexually transmitted disease control in China. PMID- 17107740 TI - Using focus groups to develop contingent valuation scenarios--a case study of women's groups in rural Nepal. AB - The construction of a contingent valuation (CV) scenario forms a critical component of willingness-to-pay (WTP) survey design, especially when working across diverse cultural and socio-economic settings. However, the methods used to develop CV scenarios have not been well reported in the health economics literature. This paper begins by describing how qualitative methods can be used to develop CV surveys. It then presents a case study illustrating how focus groups were used to develop a CV survey to value a women's group intervention in rural Nepal. A series of focus group discussions were conducted with three stakeholder groups. These were used to determine the most appropriate description of the good to be valued and the means by which payment would be elicited. These methods were very helpful in designing the survey tool and choosing the key attributes to describe the intervention. They also familiarised field workers with the concept of WTP. Further work of this kind will help to highlight additional advantages and limitations of qualitative approaches to survey design. PMID- 17107741 TI - Ocular motor indicators of executive dysfunction in fragile X and Turner syndromes. AB - Fragile X and Turner syndromes are two X-chromosome-related disorders associated with executive function and visual spatial deficits. In the present study, we used ocular motor paradigms to examine evidence that disruption to different neurological pathways underlies these deficits. We tested 17 females with fragile X, 19 females with Turner syndrome, and 40 females with neither disorder who comprised the comparison group. Group differences emerged for both the fragile X and Turner syndrome groups, each relative to the comparison group: Females with fragile X had deficits in generating memory-guided saccades, predictive saccades, and saccades made in the overlap condition of a gap/overlap task. Females with Turner syndrome showed deficits in generating memory-guided saccades, but not during either the predictive saccade or gap/overlap task. Females with Turner syndrome, but not females with fragile X, showed deficits in visually guided saccades and anti-saccades. These findings indicate that different brain regions are affected in the two disorders, and suggest that different pathways lead to the similar cognitive phenotypes described for fragile X and Turner syndromes. PMID- 17107742 TI - Late onset bipolar disorder associated with white matter hyperintensities: a pathophysiological hypothesis. AB - Vascular depression is, nowadays, a well-established concept in the literature. However, the possible emergence of late onset bipolar disorder in subjects with no antecedents of mood disorder or after a chronic or recurrent course of unipolar depression constitutes a poorly studied issue, despite its importance in clinical practice. Here, we present the case of a 72-year-old female patient who began to present recurrent major depressive symptoms, resistant to pharmacological treatment, from the age of 58. Three years later, she started to present phases of mania with rapid cycling features. A brain MRI scan showed prominent white matter hyperintensities (WMH). WMH are frequently found in the elderly population, but with greater burden in individuals with hypertension and cerebrovascular disease. WMH impair cortical function and damage the cerebral tissue. WMH have been associated with adult-onset bipolar disorder and late onset depression, and are linked to a worse prognosis of both conditions. The present case report highlights the possibility that vascular-related WMH may provoke late onset bipolar disorder by damaging frontolimbic circuits implicated in the pathophysiology of mania. PMID- 17107743 TI - Chitooligosaccharides in combination with interferon-gamma increase nitric oxide production via nuclear factor-kappaB activation in murine RAW264.7 macrophages. AB - A low-molecular weight chitosan (LMWC) with a molecular mass of 20 kDa and a chitooligosaccharide mixture (oligomixture) which is composed of sugars with a degree of polymerization (DP) of 1-6 were isolated from the chitosan hydrolysate. The effects of the chitosan hydrolysate, LMWC and oligomixture on the production of nitric oxide (NO) in RAW 264.7 macrophages were evaluated, and their effects on NF-kappaB activation and the gene expression of inducible NO synthase (iNOS) were further investigated. None of the tested 3 samples of hydrolysate, LMWC and oligomixture alone affected the NO production in RAW 264.7 macrophages. However, treatment of macrophages with a combination of hydrolysate/oligomixture and interferon-gamma (IFN-gamma) significantly induced NO production in a dose dependent manner, whereas a combination of LMWC and IFN-gamma inhibited NO production. These effects on NO synthesis were evidenced via regulating the iNOS gene expression. Both hydrolysate and oligomixture promoted the migration of NF kappaB into the nucleus and enhanced its DNA binding activity. MG132, a specific inhibitor of NF-kappaB, eliminated the NO synthesis in IFN-gamma plus hydrolysate/oligomixture-induced RAW264.7 macrophages. The treatment of RAW264.7 macrophages with anti-CD14, anti-TLR4, and anti-CR3 antibodies significantly blocked NO production induced by IFN-gamma plus hydrolysate/oligomixture. These results demonstrated that the oligomixture, which is the main functional component in the chitosan hydrolysate, in combination with IFN-gamma, synergistically induced NF-kappaB activation and NO production through binding with the receptors of CD14, TLR4 and CR3 in RAW264.7 macrophages. PMID- 17107744 TI - Outcome prediction for guidance of initial resuscitation protocol: Shock first or CPR first. AB - BACKGROUND: Ventricular fibrillation (VF) is treated optimally with a defibrillation shock shortly after patient collapse, but may benefit from initial cardiopulmonary resuscitation (CPR) if the shock is delayed. An objective measure of potential responsiveness to defibrillation could help decide optimal initial therapy. METHODS AND RESULTS: a new electrocardiogram (ECG) analysis algorithm was compared with response interval (call-to-shock) for prediction of patient outcome in a population of 87 VF patients in the Rochester, Minnesota area. In a retrospective analysis, both call-to-shock interval (p = 0.009) and ECG analysis (p < 0.001) predicted neurologically intact survival, with ECG analysis the stronger predictor (p = 0.034). When applied to advising initial patient treatment, ECG analysis compared favorably with the call-to-shock interval. Using a 7 min call-to-shock time criterion, 69% of patients would receive shocks first treatment using ECG analysis versus 67% using the call-to-shock interval (p = NS), 94% of survivors would retain successful shocks first treatment versus 85% (p = NS), and 48% of non-survivors receive alternate CPR-first treatment versus 45% (p = NS). Similarly, no significant differences were observed between ECG analysis and call-to-shock interval using an 8 min criterion. CONCLUSIONS: Both call-to-shock interval and a real-time ECG analysis are predictive of patient outcome. The ECG analysis is more predictive of neurologically intact survival. Moreover, the ECG analysis is dependent only upon the patient's condition at the time of treatment, with no need for knowledge of the response interval, which may be difficult to estimate at the time of treatment. PMID- 17107745 TI - Assessing slope stability in unplanned settlements in developing countries. AB - Unplanned housing in developing countries is often located on steep slopes. Frequently no building code is enforced for such housing and mains water is provided with no drainage provision. Both of these factors can be particularly significant in terms of landslide risk if, as is so often the case, such slopes lack any planned drainage provision. There is thus a need to develop a model that facilitates the assessment of slope stability in an holistic context, incorporating a wide range of factors (including surface cover, soil water topographic convergence, slope loading and point source water leakage) in order that appropriate advice can be given as to the general controls on slope stability in such circumstances. This paper outlines a model configured for this specific purpose and describes an application to a site in St. Lucia, West Indies, where there is active slope movement in an unplanned housing development on relatively steep topography. The model findings are in accord with the nature of the current failure at the site, provide guidance as to the significance of slope drainage and correspond to inferences drawn from an application of resistance envelope methods to the site. In being able to scenario test a uniquely wide range of combinations of factors, the model structure is shown to be highly valuable in assessing dominant slope stability process controls in such complex environments. PMID- 17107746 TI - Soil organic carbon decomposition and carbon pools in temperate and sub-tropical forests in China. AB - Decomposition of soil organic carbon (SOC) is a critical component of the global carbon cycle, and accurate estimates of SOC decomposition are important for forest carbon modeling and ultimately for decision making relative to carbon sequestration and mitigation of global climate change. We determined the major pools of SOC in four sites representing major forest types in China: temperate forests at Changbai Mountain (CBM) and Qilian Mountain (QLM), and sub-tropical forests at Yujiang (YJ) and Liping (LP) counties. A 90-day laboratory incubation was conducted to measure CO(2) evolution from forest soils from each site, and data from the incubation study were fitted to a three-pool first-order model that separated mineralizable soil organic carbon into active (C(a)), slow (C(s)) and resistant (C(r)) carbon pools. Results indicate that: (1) the rate of SOC decomposition in the sub-tropical zone was faster than that in the temperature zone, (2) The C(a) pool comprised approximately 1-3% of SOC with an average mean residence time (MRT) of 219 days. The C(s) pool comprised approximately 25-65% with an average MRT of 78 yr. The C(r) pool accounted for approximately 35-80% of SOC, (3) The YJ site in the sub-tropical zone had the greatest C(a) pool and the lowest MRT, while the QLM in the temperature zone had the greatest MRT for both the C(a) and C(s) pools. The results suggest a higher capacity for long-term C sequestration as SOC in temperature forests than in sub-tropical forests. PMID- 17107748 TI - Immunocytology in the assessment of patients with asymptomatic microhaematuria. AB - INTRODUCTION: Discrimination between malignant and nonmalignant conditions remains the key problem in assessing microhaematuria. This prospective study investigated the role of immunocytology in the evaluation of patients with microhaematuria. METHODS: uCyt+ is a commercially available immunocytologic assay based on microscopic detection of tumour-associated antigens on the membrane of urothelial cells by immunofluorescence. Between October 2000 and August 2005, 189 consecutive patients with newly diagnosed painless microhaematuria without prior transitional cell carcinoma were included. All urine samples were examined cytologically and immunocytologically. Of the 189 samples, 178 (94%) were assessable. RESULTS: Clinical assessment by physical examination, laboratory tests, endoscopy, and imaging modalities found bladder cancer in 8 patients (4%). Further diagnoses were benign prostatic hyperplasia (54 cases, 29%), cystitis (including interstitial cystitis; 20 cases, 11%), urolithiasis (18 cases, 9%), tumours of other origin (6 cases, 2%), and "further conditions" (26 cases, 13%). In 57 patients (30%) the reasons for haematuria were not disclosed. Immunocytology was positive in 7 of 8 bladder tumours (87%) and negative in 154 of 170 patients with haematuria for other reasons (91%). CONCLUSIONS: The high sensitivity and good specificity of immunocytology in the diagnosis of bladder cancer was confirmed in this population with a low disease prevalence. Only one tumour of low malignant potential was missed by immunocytology. If assessment of these patients would have been based only on immunocytology, 154 costly and invasive diagnostic procedures could have been avoided, with only 16 of 170 individuals (9%) undergoing these examinations unnecessarily. The findings justify a prospective investigation of this issue. PMID- 17107749 TI - Interleukin 8 and anti-chlamydia trachomatis mucosal IgA as urogenital immunologic markers in patients with C. trachomatis prostatic infection. AB - OBJECTIVE: This study evaluates the role of interleukin 8 (IL-8) and anti Chlamydia trachomatis (CT) immunoglobulin A (IgA) in total ejaculate (TE) of patients affected by chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) as potential markers in prostatic CT infection. METHODS: Seventy-eight consecutive patients with a diagnosis of CP/CPPS and CT infection were enrolled; 20 healthy volunteers represented the control group. All subjects underwent microbiologic analysis for common bacteria, yeasts, and viruses in TE, expressed prostatic secretion, and urine samples and molecular analysis for CT identification, anti-CT species-specific IgA, and IL-8 levels. Questionnaires regarding symptoms were given to each subject to determine correlations between clinical and laboratory data. RESULTS: Thirty-five patients were positive for CT plasmid DNA, but none of the controls were positive. Mucosal IgA was detected in 69.2% of patients and significant levels of IL-8 were detected in 75.6% of them. Significant correlations between IL-8 and mucosal IgA (p<0.001) and between IL-8 levels and symptom score results (p<0.001) were found. IL-8 values strongly correlated with CP/CPPS (p<0.001). Moreover, the patients with higher levels of IL-8 and higher positivity for IgA reported the worst symptoms. CONCLUSIONS: Our results clearly highlight the role of immune system activation in the pathophysiology of CP/CPPS and that seminal IL-8 and mucosal IgA levels specific to CT antigens appear to be the best immunologic markers of chronic chlamydial prostatitis status. PMID- 17107750 TI - Fly paper technique: a modified expansion method for preparation of postage stamp autografts by Su-Shin Lee, Tsai-Ming Lin, et al. Burns 31 (2005) p. 753. PMID- 17107751 TI - Aggregation of adverse behaviors and its affecting factors among young military conscripts in Taiwan. AB - The authors studied the prevalence of the aggregation in common lifestyle habits, namely, cigarette smoking, alcohol drinking, and betel-nut chewing and the demographic correlates of individual aggregation in these lifestyle behaviors among young military conscripts in Taiwan. Cross-sectional screening was conducted among conscripts in southern and eastern sections of Taiwan from Aug. 1st to Dec. 31st 2001. Totally, 3913 conscripts who had more than 1 month of service were included in this multistage sampling study. Information on smoking, drinking, and betel-nut chewing habits were ascertained as part of a self administered questionnaire completed by examinees at the service unit. Aggregation in lifestyle habits was studied by comparing the observed and expected proportions (O/E ratio) with their 95% confidence intervals (CI) for zero, one, two, and three simultaneously occurring lifestyle habits. The study results showed a significant clustering of lifestyle habits studied; the number of subjects was greater than expected in groups with two (for cigarette smoking and betel-nut chewing, O/E ratio=1.17, 95%CI=1.06-1.28), and three (O/E ratio=5.63, 95%CI=5.06-6.20) lifestyle habits. Determinants for this clustering of lifestyle habits included lower educational levels and residential area in southern and eastern sections of Taiwan. There was a significant individual aggregation in lifestyle habits including cigarette smoking, alcohol drinking, and betel-nut chewing in the health survey among young military conscripts. In addition, young military conscripts with low educational levels and residential area in southern and eastern sections of Taiwan had an apparent tendency toward the aggregation in these lifestyle habits. PMID- 17107752 TI - Glucocorticoids may initiate Alzheimer's disease: a potential therapeutic role for mifepristone (RU-486). AB - Alzheimer's disease (AD) is a relentless neurodegenerative disease of uncertain etiology affecting millions worldwide. Stress is an important etiological factor associated with AD. Stress and glucocorticoids (GCs) are intimately related; so much so that stress is considered to be "a condition that is able to liberate GCs". Normally, GCs are required for various bodily functions but their excess is deleterious. Hippocampus has highest density of GC receptors in brain, is particularly vulnerable to their damaging effects and undergoes reversible atrophy under their influence. Hippocampal atrophy is an initial event for the development of AD, the most common form of dementia. Apart from atrophy, GCs are able to produce a variety of other initial structural and functional changes in hippocampus. For example, these down regulate GC receptors, leading to disruption in negative feedback loop, alter dendtritic morphology and impair axonal transport. Impaired axon transport is probably an initial event that leads to the formation of paired helical filaments. Additionally, they inhibit insulin degrading enzyme, which degrades A-beta; consequently reducing its clearance. Since GCs mediate a number of initial events in AD pathogenesis, therefore GC antagonists (e.g. mifepristone) can be tried. Mifepristone has intrinsic neuroprotective and antioxidant potential which could offer additional benefits as well. Use of this drug therefore, in those with mild AD or with milder cognitive impairment can be useful. Appropriate dose, duration, safety and efficacy need to be worked out. PMID- 17107753 TI - A potential therapeutic strategy for inhibition of corneal neovascularization with new anti-VEGF agents. AB - The factors triggering corneal neovascularization involve various growth factors. The data supporting a causal role for vascular endothelial growth factor (VEGF) in corneal neovascularization are extensive. One possible strategy for treating corneal neovascularization is to inhibit VEGF activity by competitively binding VEGF with a specific neutralizing anti-VEGF antibody. The vireo-retinal service in the recent years enjoyed a high level of success in managing choroidal neovascularization using anti-VEGF strategies. Efficacy and tolerability have been demonstrated for drugs targeting VEGF. We herein hypothesize that topical application of new anti-VEGF agents such as pegaptanib, ranibizumab and bevacizumab are potentially useful for inhibiting corneal neovascularization and restoration of corneal clarity. Further investigations are needed to place these medical treatments alongside corneal neovascularization therapeutics. PMID- 17107754 TI - Could Parkinson's disease be diagnosed at an early stage by measuring rest tremor under stressed conditions? PMID- 17107755 TI - Enhancing tumor radiosensitivity by intracellular delivery of survivin antagonists. AB - Radiotherapy is frequently applied to control local tumors by mechanisms of direct killing tumor cells and inducing tumor vascular endothelial cells apoptosis. Recently, it has been demonstrated that survivin, an intracellular molecule with anti-apoptotic function, is widely expressed in human malignancies and its expression correlates with radioresistance in several tumors. Moreover, VEGF, which is highly expressed in solid tumors and further up-regulated by irradiation, has been shown to induce survivin expression in both tumor cells and vascular endothelial cells. Thus provide a survival signal to these cells and induce radioresistance to the subsequent irradiation exposure. Knocking down the expression of survivin by RNA interference or transfecting with a gene coding for a dominant negative survivin has been proved to be efficient in enhancing tumor cell radiosensitivity and improving tumor response to radiotherapy. The development of protein transduction technology made it possible to deliver large molecules into mammalian cells. We postulate that dominant negative mutants of survivin could fuse with protein transduction domain and the fusion proteins could cross cellular membranes and generate their biological activity to serve as tumor radiosensilizers. If the hypothesis proved to be practical, it would provide us an alternate method to enhance tumor radiosensitivity and the fusion proteins would be widely applicated in clinical settings because they were safer than gene therapy. PMID- 17107756 TI - Upregulation of aspartoacylase seen in diabetes is due to advanced glycation end products. PMID- 17107757 TI - A proposed solution to the clinical assessment of sarcoidosis: the sarcoidosis three-dimensional assessment instrument (STAI). AB - The clinical assessment of sarcoidosis has been confounded by its inexact diagnostic criteria, multiorgan involvement, and effects of therapy. In this manuscript an instrument, the Sarcoidosis Three-Dimensional Assessment Instrument (STAI), is proposed to assess the clinical state of sarcoidosis. The instrument examines each organ involved with sarcoidosis separately. For each organ, the instrument contains three axes: involvement, severity, and activity. Involvement is based upon a previously described instrument that has been updated to capture involvement of more organs and to account for advances in diagnostic testing for sarcoidosis. Severity is based both on the decline from normal capacity as well as physical and psychosocial limitation. Disease activity takes into account changes in organ function as well as changes in therapy. Although this instrument is presently not validated, it is hoped that it will undergo study as it rationally accounts for several problems of previous assessment instruments. PMID- 17107758 TI - Antiinflammatory and analgesic activities of Zizyphus lotus root barks. AB - The root barks of Zizyphus lotus were extracted with water, chloroform, ethyl acetate and methanol to determine their antiinflammatory and analgesic activities. Aqueous extract (50, 100 and 200 mg/kg) given intraperitoneally (i.p.) showed a significant and dose-dependent antiinflammatory and analgesic activity. PMID- 17107760 TI - Avian influenza; routes of transmission: lessons and thoughts drawn out of the past and present situation in the world and in the European Union. PMID- 17107759 TI - Alpha-bulnesene, a PAF inhibitor isolated from the essential oil of Pogostemon cablin. AB - Steam distilled essential oil from the aerial parts of Pogostemon cablin was analyzed by gas chromatography-mass spectrometry (GC-MS). Forty one compounds were identified of which alpha-guaiene (20.62%) and alpha-bulnesene (16.18%) were major constituents. Furthermore, fractionation of the essential oil from P. cablin guided by inhibitory activity against PAF-induced platelet aggregation led to the isolation of the sesquiterpene, alpha-bulnesene. PMID- 17107761 TI - [A high troponin value: about a favourable evolution case]. AB - Cardiac troponin I (TnIC) is a sensitive and specific marker for myocardial injuries. A part from its diagnosis character, troponin is a major element for mid term prognosis with regard to occurred cardiovascular events. We are reporting the case of a 56-year-old man admitted to hospital for an inaugural myocardial infarction with positive evolution despite a very high level of troponin (1200 ng/ml) in post re-vascularisation. The context of moderate risk factors and the early re-vascularisation probably contributed to a favourable evolution of the patient. A very high troponin value seems to be more in favour of an effective re-perfusion rather than a mid term prognosis factor in this case. PMID- 17107762 TI - Molecular characterization and expression analysis of nine cotton GhEF1A genes encoding translation elongation factor 1A. AB - The translation elongation factor 1A, eEF1A, plays an important role in protein synthesis, catalyzing the binding of aminoacyl-tRNA to the A-site of the ribosome by a GTP-dependent mechanism. To investigate the role of eEF1A for protein synthesis in cotton fiber development, nine different cDNA clones encoding eukaryotic translation elongation factor 1A were isolated from cotton (Gossypium hirsutum) fiber cDNA libraries. The isolated genes (cDNAs) were designated cotton elongation factor 1A gene GhEF1A1, GhEF1A2, GhEF1A3, GhEF1A4, GhEF1A5, GhEF1A6, GhEF1A7, GhEF1A8, GhEF1A9, respectively. They share high sequence homology at nucleotide level (71-99% identity) in the coding region and at amino acid level (96-99% identity) among each other. Phylogenetic analysis demonstrated that the nine GhEF1A genes can be divided into 5-6 subfamilies, indicating the divergence occurred in structures of the genes as well as the deduced proteins during evolution. Real-time quantitative RT-PCR analysis revealed that GhEF1A genes are differentially expressed in different tissues/organs. Of the nine GhEF1A genes, five are expressed at relatively high levels in young fibers. Further analysis indicated that expressions of the GhEF1As in fiber are highly developmental regulated, suggesting that protein biosynthesis is very active at the early fiber elongation. PMID- 17107763 TI - Life without GAG: the BARE-2 retrotransposon as a parasite's parasite. AB - A large proportion of the plant LTR (Long Terminal Repeat) retrotransposons are partly or completely unable to synthesize their own machinery for transposition. However, most of these inactive or non-autonomous elements are likely able to retrotranspose, based on their insertional polymorphism. Therefore, they must be parasitic on one or more active partners. Here, we describe the parasitism of the chimeric BARE-2 element on the active BARE-1 (Barley RetroElement-2 and -1 respectively). These two elements are present in the Triticeae and related species, and are together polymorphic among closely related accessions. BARE-2 elements are unable to synthesize their own GAG protein, and harbor a specific ATG deletion in the gag ORF. However, BARE-2 sequences are conserved with BARE-1 in the PBS (Primer Binding Site), PSI (Packaging SIgnal) and DIS (DImerization Signal) domains. As these motifs have been shown to allow parasitism among the lentiviruses, we conclude that BARE-2 is probably a partial parasite of the BARE 1 element because the machinery of the latter can complement the defective GAG of the former. This example emphasizes that we must characterize the parasitic network of LTR retrotransposons and its implication for integration of autonomous, inactive, and non-autonomous elements in order to understand current and past host genome evolution. PMID- 17107764 TI - Cloning, genomic organisation and mRNA expression of a pectin lyase gene from a mutant strain of Penicillium occitanis. AB - The regulatory cis elements of fungal pectinases are well studied in Aspergillus genera but little is known in other fungal species. A genomic bank from Penicillium occitanis fungus is constructed and screened by previously isolated cDNA probe of a pectin lyase. From several isolated clones, the nucleotide sequence of the pectin lyase gene was completed and led to the identification of introns and promoter-terminator regions. A streaking future was found in pnl gene of P. occitanis: it exhibits the highest nucleotide homology with the pnlA of Aspergillus niger but the positions of its 4 introns is completely identical to that of A. niger pnlB gene. In addition to the determination of transcription start site, the promoter sequence from the pnl gene was analysed. It showed the conservation of known consensus sequences -CreA, Hap2-3-4, PacC ...-, and the existence of a particular sequence -CCTGA- which is similar to that already found to be specific of pectinolytic gene in Aspergillus, CCCTGA. This result suggests that the corresponding regulatory trans-acting factor should be the same as in Aspergillus. PMID- 17107765 TI - Preparation, characterization and controlled release investigation of interpenetrating polymer networks of poly(acrylic acid)/triazole modified poly(vinyl alcohol). AB - A series of interpenetrating polymer networks of poly(acrylic acid) (PAA)/triazole modified poly(vinyl alcohol) (TMIPNs) were synthesized by radical polymerization in methanol at room temperature with l-ascorbic acid (Vc) and peroxide hydrogen (H2O2) as initiators and trihydroxymethyl propane glycidol ether (6360) as a crosslinker. The structures of the gels were characterized by Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The swelling/deswelling behavior of hydrogels was studied in different pH and different concentrations of NaCl aqueous solutions. The results showed that the TMIPNs hydrogels had excellent pH- and salt-sensitivity in the range of the investigation. The mechanism of the swelling and the deswelling was discussed and the results were confirmed further by scanning electron microscope (SEM). In addition, the controlled release behavior of TMIPNs in vitro was also studied. The effects of physical stimulus (ultraviolet ray and ultrasonic wave), salt concentration, pH value and the swelling/deswelling on the controlled released behavior were also explored. PMID- 17107766 TI - Temporal and genetic influences on protection against noise-induced hearing loss by hypoxic preconditioning in mice. AB - The protective benefits of hypoxic preconditioning (HPC) against permanent noise induced hearing loss (NIHL) were investigated in mice. Hypoxia induced by exposure to 8% O2 for 4 h conferred significant protection against damaging broadband noise delivered 24-48 h later in male and female CBA/J (CBA) and CBA/CaJ mice. No protection was found in C57BL/6 (B6) mice, their B6.CAST Cdh23(CAST) (B6.CAST) congenics, or in CBAxB6 F1 hybrid mice over the same interval, suggesting that the potential for HPC depends on one or a few autosomal recessive alleles carried by CBA-related strains, and is not influenced by the Cdh23 locus. Protection against NIHL in CBA mice was associated with significant up-regulation of hypoxia-inducible factor-1alpha (HIF-1alpha) within the organ of Corti, not found in B6.CAST. In both CBA and B6.CAST mice, some hypoxia-noise intervals shorter than 24 h were associated with exacerbation of NIHL. Cellular cascades underlying the early exacerbation of NIHL by hypoxia are therefore common to both strains, and not mechanistically linked to later protection. Elucidation of the events that underlie HPC, and how these are impacted by genetics, may lead to pharmacologic approaches to mimic HPC, and may help identify individuals with elevated risk of NIHL. PMID- 17107767 TI - Binaural pitch perception in normal-hearing and hearing-impaired listeners. AB - The effects of hearing impairment on the perception of binaural-pitch stimuli were investigated. Several experiments were performed with normal-hearing and hearing-impaired listeners, including detection and discrimination of binaural pitch, and melody recognition using different types of binaural pitches. For the normal-hearing listeners, all types of binaural pitches could be perceived immediately and were musical. The hearing-impaired listeners could be divided into three groups based on their results: (a) some perceived all types of binaural pitches, but with decreased salience or musicality compared to normal hearing listeners; (b) some could only perceive the strongest pitch types; (c) some were unable to perceive any binaural pitch at all. The performance of the listeners was not correlated with audibility. Additional experiments investigated the correlation between performance in binaural-pitch perception and performance in measures of spectral and temporal resolution. Reduced frequency discrimination appeared to be linked to poorer melody recognition skills. Reduced frequency selectivity was also found to impede the perception of binaural-pitch stimuli. Overall, binaural-pitch stimuli might be very useful tools within clinical diagnostics for detecting specific deficiencies in the auditory system. PMID- 17107768 TI - Influence of lifetime food restriction on physiological variables in Labrador retriever dogs. AB - Effects of lifetime food restriction on erythrocytes and numerous clinical chemistry, thyroid, parathyroid, and acid-base variables are reported from a paired-feeding study of 25% diet restriction in dogs. The 48 dogs were paired by gender and weaning weight within litter, and 1 dog in each pair was fed 25% less than its pair mate, from age 8 weeks until death. Erythrocyte and serum biochemistry profiles were evaluated by annual sampling intervals and longitudinally. Erythrocyte variables were slightly higher among control-fed dogs, a possible reflection of the need to support both higher body fat mass and lean mass that uses energy less efficiently. Among serum biochemistry variables, glucose and triglyceride were lower among diet-restricted dogs, while creatinine was slightly higher in the absence of renal disease or failure, over the life spans of the dogs. Glucose outcomes reflect improved glucose tolerance that has been demonstrated with diet restriction protocols in several species, while triglyceride data may reflect the difference in total body fat cells between feeding groups. Creatinine outcomes may reflect lean mass responses to diet restriction or more efficient function of the intracellular proteasome. Serum triiodothyronine levels were lower among diet-restricted dogs. Other clinical chemistry and thyroid variables, parathyroid variables, and acid-base variables were not strongly influenced by diet restriction but revealed age-related effects. PMID- 17107769 TI - Immunogenicity of the rF1+rV vaccine for plague with identification of potential immune correlates. AB - The rF1+rV candidate sub-unit vaccine for plague, formulated by adsorption to alhydrogel, has been demonstrated to be immunogenic in the cynomolgus macaque in a clinically relevant dose-range (5-40 microg of each sub-unit) and regimen. Following two doses of vaccine, a specific IgG titre developed in a dose-related manner with predominance of the IgG1/IgG2 isotypes. Groups of macaques receiving only a single dose of vaccine at the 40 microg dose-level had a significantly reduced peak IgG response and faster decline to baseline. Serum collected at week 5 from 19 immunised animals competed with and displaced murine Mab7.3 from binding to the V antigen in vitro. By week 53 of the schedule, although absolute IgG titres had declined, 17/19 macaque sera tested contained competing antibody, indicating the durability of a functional immune response to rF1+rV in this species. Thirteen of these week 53 sera were passively transferred into groups of naive mice, and all conferred full or partial protection against subsequent challenge of the mice with plague. Generally, those sera which were most competitive with Mab 7.3 for binding to V antigen were fully protective by passive transfer, although one week-53 serum sample was fully protective by passive transfer but not active by competitive ELISA. The early development of protective immunity in macaques was also indicated from the protection conferred on naive mice by the passive transfer of immune macaque serum collected at 2-10 weeks of the immunisation schedule. Serum samples from representative macaques within this time period also inhibited the Yersinia-mediated cytotoxicity of J774 macrophages in a qualitative in vitro assay of type three secretion. PMID- 17107770 TI - Captopril ameliorates toxicity induced by paraquat in mitochondria isolated from the rat liver. AB - The aim of the present study was to show the abilities of captopril as a thiol ACEi (angiotensin converting enzyme inhibitor), on mitochondria toxicity due to paraquat. Mitochondrial isolation from rat liver was divided into 4 groups. Group 1 was considered as control, group 2 received paraquat (5 mM), group 3 received captopril (0.08 mM) and group 4 received paraquat (5 mM)+captopril (0.08 mM). Lipid peroxidation, catalase activity, GSH (reduced glutathione) and GSSG (oxidized glutathione) concentrations were determined in isolated rat liver mitochondria. Simultaneous treatment of mitochondria with captopril (0.08 mM)+paraquat (5 mM) significantly ameliorate the mitochondria toxicity induced by paraquat (5 mM) alone. The results confirm antioxidant effect of captopril. This effect appears to be attributable to the Sulfhydryl Groups (SH) in the compound which may be due to captopril abilities to scavenge reactive oxygen species. The results indicate that captopril may prevent oxidative stress induced by paraquat. PMID- 17107771 TI - Effects of red wine on ochratoxin A toxicity in intestinal Caco-2/TC7 cells. AB - Ochratoxin A (OTA) is found in a variety of foods and beverages, including red wine. OTA was reported to be nephrotoxic, immunotoxic, hepatotoxic and a potential carcinogen, with yet uncharacterized mechanisms. Consumption of contaminated wines might contribute up to 13% of OTA daily human intake. Potentially chronic exposure has therefore raised public health concern. OTA toxicity in the presence of de-alcoholated red wine was investigated in human intestinal Caco-2/TC7 cells, differentiated on filter supports, by measuring tight junction (TJ) permeability, morphological alterations of TJ proteins and occurrence of apoptosis. Cells were treated with OTA, in the presence of de alcoholated red wine, for 48h and the ability to recover from the effects of OTA was evaluated after 24h in complete medium. OTA treatment increased TJ permeability and caused intracellular redistribution of claudin-4. However, cells were able to restore permeability and correct localization of claudin-4 following 24h recovery. Conversely, in the presence of red wine, OTA produced faster and irreversible increase in TJ permeability, intracellular delocalization of claudin 4 and extensive apoptosis. Our results point at a possible synergy between OTA and some red wine components, such as polyphenols, in the induction of apoptotic cell death. PMID- 17107772 TI - The pro-inflammatory oxidant hypochlorous acid induces Bax-dependent mitochondrial permeabilisation and cell death through AIF-/EndoG-dependent pathways. AB - At sites of chronic inflammation, such as in the inflamed rheumatoid joint, activated neutrophils release hydrogen peroxide (H(2)O(2)) and the enzyme myeloperoxidase to catalyse the formation of hypochlorous acid (HOCl). 3 chlorotyrosine, a marker of HOCl in vivo, has been observed in synovial fluid proteins from rheumatoid arthritis patients. However the mechanisms of HOCl induced cytotxicity are unknown. We determined the molecular mechanisms by which HOCl induced cell death in human mesenchymal progenitor cells (MPCs) differentiated into a chondrocytic phenotype as a model of human cartilage cells and show that HOCl induced rapid Bax conformational change, mitochondrial permeability and release of intra-mitochondrial pro-apoptotic proteins which resulted in nuclear translocation of AIF and EndoG. siRNA-mediated knockdown of Bax substantially prevented mitochondrial permeability, release of intra mitochondrial pro-apoptotic proteins. Cell death was inhibited by siRNA-mediated knockdown of Bax, AIF or EndoG. Although we observed several biochemical markers of apoptosis, caspase activation was not detected either by western blotting, fluorescence activity assays or by using caspase inhibitors to inhibit cell death. This was further supported by findings that (1) in vitro exposure of recombinant human caspases to HOCl caused significant inhibition of caspase activity and (2) the addition of HOCl to staurosporine-treated MPCs inhibited the activity of cellular caspases. Our results show for the first time that HOCl induced Bax-dependent mitochondrial permeability which led to cell death without caspase activity by processes involving AIF/EndoG-dependent pathways. Our study provides a novel insight into the potential mechanisms of cell death in the inflamed human joint. PMID- 17107773 TI - Emotional prosodic processing in auditory hallucinations. AB - Deficits in emotional prosodic processing, the expression of emotions in voice, have been widely reported in patients with schizophrenia, not only in comprehending emotional prosody but also expressing it. Given that prosodic cues are important in memory for voice and speaker identity, Cutting has proposed that prosodic deficits may contribute to the misattribution that appears to occur in auditory hallucinations in psychosis. The present study compared hallucinating patients with schizophrenia, non-hallucinating patients and normal controls on an emotional prosodic processing task. It was hypothesised that hallucinators would demonstrate greater deficits in emotional prosodic processing than non hallucinators and normal controls. Participants were 67 patients with a diagnosis of schizophrenia or schizoaffective disorder (hallucinating=38, non hallucinating=29) and 31 normal controls. The prosodic processing task used in this study comprised a series of semantically neutral sentences expressed in happy, sad and neutral voices which were rated on a 7-point Likert scale from sad (-3) through neutral (0) to happy (+3). Significant deficits in the prosodic processing tasks were found in hallucinating patients compared to non hallucinating patients and normal controls. No significant differences were observed between non-hallucinating patients and normal controls. In the present study, patients experiencing auditory hallucinations were not as successful in recognising and using prosodic cues as the non-hallucinating patients. These results are consistent with Cutting's hypothesis, that prosodic dysfunction may mediate the misattribution of auditory hallucinations. PMID- 17107774 TI - State and trait related predictors of serum cortisol to DHEA(S) molar ratios and hormone concentrations in schizophrenia patients. AB - OBJECTIVE: In previous studies we have demonstrated high serum molar ratios of cortisol to dehydroepiandrosterone (DHEA) and its sulfate ester (DHEAS) [together abbreviated DHEA(S)], and the value of both cortisol/DHEA(S) molar ratios for prediction of responsivity to antipsychotic treatment in schizophrenia patients. The present study aimed to examine the contribution of anxiety, and severity of symptoms to the prediction of serum cortisol, DHEA(S) levels and two molar ratios across three examinations. METHOD: Serum concentrations of cortisol and DHEA(S)were examined in 43 schizophrenia inpatients and in 20 age matched healthy controls at baseline, and after 2 and 4 weeks. The Positive and Negative Symptom Scale and the State-Trait Anxiety Inventory scores were used as independent variables for multiple regression analysis. RESULTS: Despite clinical improvement during the study period cortisol/DHEA(S) molar ratios were found persistently elevated as compared to healthy controls. Multiple regression analysis revealed that across three examinations cortisol/DHEA(S) molar ratios negatively associated with trait-anxiety (partial R(2)=7-14%) rather than with negative symptoms (partial R(2)=3-6%). Age and age of onset account for 12.7% for variability of cortisol/DHEAS ratio. Serum cortisol concentrations are predicted by trait and state-anxiety, activation symptoms and daily doses of antipsychotics. A small portion of variability in serum DHEA levels (R(2)=9%) is associated with symptom severity, while DHEAS levels were predicted by age at examination and age of onset. CONCLUSION: Elevated serum cortisol/DHEA(S) molar ratios were attributed to trait-anxiety and age rather than to clinical symptoms. The findings may indicate persistent dysfunction of the hypothalamic-pituitary adrenal axis that is independent of the patients' clinical state. PMID- 17107775 TI - Developmental and hormonal regulation of midgut remodeling in a lepidopteran insect, Heliothis virescens. AB - Midgut tissue undergoes remodeling during metamorphosis in insects belonging to orders Lepidoptera and Diptera. We investigated the developmental and hormonal regulation of these remodeling events in lepidopteran insect, Heliothis virescens. In H. virescens, programmed cell death (PCD) of larval midgut cells as well as proliferation and differentiation of imaginal cells began at 108 h after ecdysis to the final larval instar (AEFL) and proceeded through the pupal stages. Expression patterns of pro- cell death factors (caspase-1 and ICE) and anti-cell death factor, Inhibitor of Apoptosis (IAP) were studied in midguts during last larval and pupal stages. IAP, Caspase-1 and ICE mRNAs showed peaks at 48 h AEFL, 96 h AEFL and in newly formed pupae, respectively. Immunohistochemical analysis substantiated high caspase-3 activity in midgut at 108 h AEFL. Application of methoprene, a juvenile hormone analog (JHA) blocked PCD by maintaining high levels of IAP, downregulating the expression of caspase-1, ICE and inhibiting an increase in caspase-3 protein levels in midgut tissue. Also, the differentiation of imaginal cells was impaired by methoprene treatment. These studies demonstrate that presence of JHA during final instar larvae affects both midgut remodeling and larval-pupal metamorphosis leading to larval/pupal deformities in lepidopteran insects, a mechanism that is different from that in mosquito, Ae. aegypti where JHA uncouples midgut remodeling from metamorphosis. PMID- 17107777 TI - High throughput screening of protein formulation stability: practical considerations. AB - The formulation of protein drugs is a difficult and time-consuming process, mainly due to the complexity of protein structure and the very specific physical and chemical properties involved. Understanding protein degradation pathways is essential for the success of a biopharmaceutical drug. The present review concerns the application of high throughput screening techniques in protein formulation development. A protein high throughput formulation (HTF) platform is based on the use of microplates. Basically, the HTF platform consists of two parts: (i) sample preparation and (ii) sample analysis. Sample preparation involves automated systems for dispensing the drug and the formulation ingredients in both liquid and powder form. The sample analysis involves specific methods developed for each protein to investigate physical and chemical properties of the formulations in microplates. Examples are presented of the use of protein intrinsic fluorescence for the analysis of protein aqueous properties (e.g., conformation and aggregation). Different techniques suitable for HTF analysis are discussed and some of the issues concerning implementation are presented with reference to the use of microplates. PMID- 17107776 TI - Differential Delta expression underlies the diversity of sensory organ patterns among the legs of the Drosophila adult. AB - Many studies have shown that morphological diversity among homologous animal structures is generated by the homeotic (Hox) genes. However, the mechanisms through which Hox genes specify particular morphological features are not fully understood. We have addressed this issue by investigating how diverse sensory organ patterns are formed among the legs of the Drosophila melanogaster adult. The Drosophila adult has one pair of legs on each of its three thoracic segments (the T1-T3 segments). Although homologous, legs from different segments have distinct morphological features. Our focus is on the formation of diverse patterns of small mechanosensory bristles or microchaetae (mCs) among the legs. On T2 legs, the mCs are organized into a series of longitudinal rows (L-rows) precisely positioned along the leg circumference. The L-rows are observed on all three pairs of legs, but additional and novel pattern elements are found on T1 and T3 legs. For example, at specific positions on T1 and T3 legs, some mCs are organized into transverse rows (T-rows). Our studies indicate that the T-rows on T1 and T3 legs are established as a result of Hox gene modulation of the pathway for patterning the L-row mC bristles. Our findings suggest that the Hox genes, Sex combs reduced (Scr) and Ultrabithorax (Ubx), establish differential expression of the proneural gene achaete (ac) by modifying expression of the ac prepattern regulator, Delta (Dl), in T1 and T3 legs, respectively. This study identifies Dl as a potential link between Hox genes and the sensory organ patterning hierarchy, providing insight into the connection between Hox gene function and the formation of specific morphological features. PMID- 17107778 TI - Patient experiences with pulmonary fibrosis. AB - This survey describes the experiences of patients diagnosed with pulmonary fibrosis, focusing on the issues of patient education and resources. A survey of 52 defined-choice and open-ended questions regarding the diagnosis and management of pulmonary fibrosis was delivered. A total of 1448 respondents comprised the study group. Two-thirds of respondents reported a clear lack of information and resources on pulmonary fibrosis at the time of diagnosis. Less than half of respondents reported they felt well-informed about treatment options, the role of supplemental oxygen, pulmonary rehabilitation, and transplantation. These results suggest there is a substantial need for improved patient education regarding the diagnosis and management of pulmonary fibrosis. PMID- 17107779 TI - Home mechanical ventilation. A growing challenge in an aging society. PMID- 17107780 TI - Performance and emission study of biodiesel from leather industry pre-fleshings. AB - The possible use of pre-fleshing wastes from tanneries in the production of biodiesel fuel was examimed in our previous study in 2005. In this study, engine performance and emission values of biodiesel obtained from these wastes have been investigated and compared to petrodiesel fuel. The test results demonstrated that although the wheel force and power of biodiesel fuel is slightly lower than petrodiesel and the acceleration periods are accordingly longer, the emission values of hydrocarbons and particulate matter were found to be considerably lower with biodiesel. Thus, it has been shown that a waste byproduct can be utilized in the production of an environmentally friendly fuel, which can be used in some diesel engines without a need for major adjustment, providing economic and ecological benefits. PMID- 17107781 TI - Food for thought - should we stop doing research? PMID- 17107782 TI - The weakest link. PMID- 17107783 TI - Feasibility of hydrogen production in thermophilic mixed fermentation by natural anaerobes. AB - The biological sludge from an animal wastewater treatment plant was treated to enrich hydrogen-producing mixed bacteria, and effects on hydrogen yield were investigated during anaerobic fermentation at 55 degrees C. Enrichment of hydrogen-producing bacteria was conducted at pH adjustment of inocula to 3 and 5 with and without additional heat treatment (NHT and HT). The enriched mixed bacteria were cultivated at initial pHs of 5, 6, and 7 with synthetic organic wastewater containing different levels of nitrogen (2.0 and 0.8 g/l as total nitrogen) under static batch conditions. The main effects of heat treatment and enrichment pH were significant on hydrogen production. There was no significant effect of different nitrogen concentrations on hydrogen production. The methane free biogas contained hydrogen levels of up to 64% for a fermentative condition that showed maximum hydrogen evolution (at culture pH 5 after enrichment at pH 5 with HT). The dominating intermediate metabolites were acetate, n-butyrate, and ethanol. Yields of produced hydrogen were significantly dependent upon levels of n-butyrate. PMID- 17107784 TI - Microbially-influenced degradation of solidified/stabilized metal waste. AB - In the present study, a refined microbially-influenced degradation method was used to evaluate the stability of a solidified synthetic waste containing chromium salt, cement and fly ash in two different proportions. The experimental samples showed evidence of microbial growth by leaching of sulfate. Chromium leached by Thiobacillus thiooxidans from the experimental samples 'C1' (10.26% CrCl3 .6H2O; 89.74% cement) and 'FC1' (10.26% CrCl3 .6H2O; 10% fly ash; 79.74% cement), after 30 days of exposure was 14.53 mg/g and 9.53 mg/g, respectively. The corresponding concentration of chromium in the leachate was 0.189 mg/l and 0.124 mg/l, respectively, which was lower than the toxicity characteristic leaching procedure (TCLP), regulatory limit (5 mg/l). Replacement of cement by 10% fly ash in FC1 restricted the leaching of chromium more effectively. Model equations based on two shrinking core models namely, acid dissolution and bulk diffusion model, were used to analyze the kinetics of microbial degradation. Of the two approaches, the bulk diffusion model fit the data better than the acid dissolution model as indicated by the correlation coefficients of >0.97. PMID- 17107785 TI - Determination of bulk mass transfer coefficient of biosorption on sludge granule based on liquid membrane mass transfer mechanism. AB - A bulk mass transfer coefficient (BMTC) equation was derived from the mechanism of mass transfer in surface liquid membrane in this study, which was based on the analysis of biosorption process, conservation of mass in sludge granule and the unification of the dimension. A biosorption experiment was carried out in which anoxic sludge from an anoxic baffled reactor for printing and dyeing wastewater treatment was used to adsorb Acid Red GR dye. The results showed that there was a linear regression curve between ln[qe/(qe-q)] (qe and q were the amount adsorbed at equilibrium and at time t, respectively.) and time t. There was also a good agreement between the adsorbate amount measured and that predicted by the equation of BMTC. The BMTC of Acid Red GR dye adsorbed by anoxic sludge was 6.816 kgm(-3)min(-1). Experimental results indicated that the BMTC determined by a simple adsorptive experiment using this equation was credible. It could be a feasible and effective way to determine BMTC of activated sludge for biosorption performance. PMID- 17107786 TI - Influence of process conditions on end product formation from Clostridium thermocellum 27405 in solid substrate cultivation on paper pulp sludge. AB - Solid substrate cultivation of thermophilic, anaerobic bacteria offers an alternative production method for many bio-based chemicals; however the process must be optimized for each substrate-organism fermentation. The effects of initial substrate moisture content (SMC, 30%, 50% and 70% wet-basis), supplemental nutrient concentration (SNC, 12%, 50% and 100%) and duration of cultivation time (6, 10 and 14 days), on product formation (lactate, ethanol and acetate) by Clostridium thermocellum 27405 were examined during growth on paper pulp sludge. Water activities at moisture contents above 30% wet-basis were essentially identical ( approximately 0.99), yet the water contents differed significantly, and affected the metabolic activity of C. thermocellum. Increases in initial substrate moisture content from 50% to 70% for cultures supplemented with 50% or 100% nutrients resulted in a 75-145 mM increase in total end products. At 70% SMC, the addition of 100% SNC generated a 56% increase in product formation above the addition of 50% nutrient supplementation. Increases in the quantity of free water present in the solid substrate cultivation system up to the water holding capacity of the paper pulp sludge led to improved performance of this anaerobic bacterium. While nutrient supplementation is common in the form of salts for many aerobic microorganisms, efficient metabolism for anaerobic C. thermocellum grown in SSC was highly dependent on added salts, vitamins and reducing agents. Further studies are needed to determine if this is a general effect for other anaerobes grown in solid substrate cultures. PMID- 17107787 TI - Cloning, purification, and characterization of chitinase from Bacillus sp. DAU101. AB - A chitinase encoding gene from Bacillus sp. DAU101 was cloned in Escherichia coli. The nucleotide sequencing revealed a single open reading frame containing 1781 bp and encoding 597 amino acids with 66 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis and zymogram. The chitinase was composed of three domains: a catalytic domain, a fibronectin III domain, and a chitin binding domain. The chitinase was purified by GST-fusion purification system. The pH and temperature optima of the enzyme were 7.5 and 60 degrees C, respectively. The metal ions, Zn(2+), Cu(2+), and Hg(2+), were strongly inhibited chitinase activity. However, chitinase activity was increased 1.4-fold by Co(2+). Chisb could hydrolyze GlcNAc(2) to N-acetylglucosamine and was produced GlcNAc(2), when chitin derivatives were used as the substrate. This indicated that Chisb was a bifunctional enzyme, N-acetylglucosaminase and chitobiosidase. The enzyme could not hydrolyze glycol chitin, glycol chitosan, or CMC, but hydrolyzed colloidal chitin and soluble chitosan. PMID- 17107788 TI - Biosorption and bioaccumulation of microelements by Riccia fluitans in single and multi-metal system. AB - In the present study, biosorption and bioaccumulation characteristics of Riccia fluitans to be use as biological mineral feed supplement, was investigated. Preliminary studies showed that R. fluitans was rich in protein (27-31%) and possessed high cation exchange capacity (14.5 mequiv g(-1)) and therefore it has the potential to find an application as biological carrier of microelements that are supplied to feed of animals, the diet of which is deficient in these components. In the present study, various processes of enrichment with microelements of crystalwort were investigated, including biosorption, bioaccumulation by non-growing and growing cells in single-(Cr(III) ions) and multi-metal system (Cr(III), Cu(II), Mn(II), Zn(II) ions). The effect of process parameters (temperature and pH) on metal ions binding efficiency was studied in single-metal system. It was found that at 20 degrees C and pH5 the biomass bound 106 mg g(-1) Cr(III) ions. The experimental results showed that the mostly advantageous process of metal ions binding was biosorption, the process that is also the mostly cost-effective. PMID- 17107789 TI - Performance of membrane-less microbial fuel cell treating wastewater and effect of electrode distance and area on electricity production. AB - Performance of mediator-less and membrane-less microbial fuel cell (ML-MFC) was evaluated to treat synthetic wastewater and actual sewage. The ML-MFC gave COD and BOD removal efficiencies of 88% and 87%, respectively, and TKN removal was around 45-50%. Biomass granulation was observed in the anode compartment of ML MFC. Effect of distance between the electrodes and total surface area of anode on electricity production was evaluated under variable external resistance. Maximum power density of 10.9 and 10.13 mW/m2 was observed at lower spacing between the electrodes (20 cm) and for lesser surface area of the anode, respectively. With variation in the carbon source in the feed, variation in power production was observed. PMID- 17107791 TI - Aminomethyl tetrahydronaphthalene biphenyl carboxamide MCH-R1 antagonists- Increasing selectivity over hERG. AB - Aminomethyl tetrahydronaphthalene biphenyl carboxamide MCH-R1 antagonists with greater selectivity over hERG were identified. SAR studies addressing two distinct alternatives for structural modifications leading to improve hERG selectivity are described. PMID- 17107790 TI - Identification and optimisation of a series of substituted 5-pyridin-2-yl thiophene-2-hydroxamic acids as potent histone deacetylase (HDAC) inhibitors. AB - Further investigation of a series of thienyl-based hydroxamic acids that included ADS100380 and ADS102550 led to the identification of the 5-pyridin-2-yl-thiophene 2-hydroxamic acid 3c, which possessed modest HDAC inhibitory activity. Substitution at the 5- and 6-positions of the pyridyl ring of compound 3c provided compounds 5a-g, 7a, b, 9, and 13a. Compound 5b demonstrated improved potency, in vitro DMPK profile, and rat oral bioavailability, compared to ADS102550. Functionalisation of the pendent phenyl group of compounds 5b, 5e and 13a provided analogues that possessed excellent enzyme inhibition and anti proliferative activity. PMID- 17107793 TI - Evaluating scoring functions for docking and designing beta-secretase inhibitors. AB - Several simple scoring methods were examined for 2 series of beta-secretase (BACE 1) inhibitors to identify a docking/scoring protocol which could be used to design BACE-1 inhibitors in a drug discovery program. Both the PLP1 score and MMFFs interaction energy (E(inter)) performed as well or better than more computationally intensive methods for a set of substrate-based inhibitors, while the latter performed well for both sets of inhibitors. PMID- 17107792 TI - Constrained analogs of CB-1 antagonists: 1,5,6,7-Tetrahydro-4H-pyrrolo[3,2 c]pyridine-4-one derivatives. AB - A series of pyrrolopyridinones was designed and synthesized as constrained analogs of the pyrazole CB-1 antagonist rimonabant. Certain examples exhibited very potent hCB-1 receptor binding affinity and functional antagonism with Ki and Kb values below 10 nM, and with high selectivity for CB-1 over CB-2 (>100-fold). A representative analog was established to cause significant appetite suppression and reduction in body weight gain in industry-standard rat models used to develop new therapeutics for obesity. PMID- 17107794 TI - MCH-R1 antagonists based on an arginine scaffold: SAR studies on the amino terminus. AB - We have identified a novel series of potent MCH-R1 antagonists based on l arginine. As predicted by computational methods, there was an activity dependence on the pi-electronic character of the aromatic systems corresponding to the amino terminus of these molecules. These results have enhanced our understanding of the MCH-R1 receptor and the potential for a predictive homology model. PMID- 17107795 TI - PDE5 inhibitors: An original access to novel potent arylated analogues of tadalafil. AB - A method to access totally new analogues of tadalafil was explored. The Buchwald reaction was adapted and used to replace the methyl group of tadalafil by various aryl groups. Inhibition potencies on PDE5 of these analogues were determined and proved to be comparable to the one of tadalafil. Using the same route, compounds with the same level of activity but improved water solubility were produced by introducing a pyridine or a pyrimidine ring. This original route also opens access to new unpatented compounds. PMID- 17107796 TI - Aminomethyl tetrahydronaphthalene ketopiperazine MCH-R1 antagonists--Increasing selectivity over hERG. AB - A direct correlation between hERG binding and QTc prolongation was established for a series of aminomethyl tetrahydronaphthalene ketopiperazine MCH-R1 antagonists. Compounds within this class with greater selectivity over hERG were developed. Compound 4h proved to have the best profile, with MCH-R1 Ki = 16 nm and hERG IC50 = 25 microM. PMID- 17107797 TI - Discovery of a new class of 4-anilinopyrimidines as potent c-Jun N-terminal kinase inhibitors: Synthesis and SAR studies. AB - A new series of 4-anilinopyrimidines has been synthesized and evaluated as JNK1 inhibitors. SAR studies led to the discovery of potent JNK1 inhibitors with good enzymatic activity as well as cellular potency represented by compound 2b. Kinase selectivity profile and the crystal structure of 2b are also described. PMID- 17107798 TI - Dual serotonin transporter/histamine H3 ligands: Optimization of the H3 pharmacophore. AB - A series of tetrahydroisoquinolines acting as dual histamine H3/serotonin transporter ligands is described. A highly regio-selective synthesis of the tetrahydroisoquinoline core involving acid mediated ring-closure of an acetophenone intermediate followed by reduction with NaCNBH3 was developed. In vitro and in vivo data are discussed. PMID- 17107799 TI - From dihydroxypyrimidine carboxylic acids to carboxamide HIV-1 integrase inhibitors: SAR around the amide moiety. AB - 4,5-Dihyroxypyrimidine carboxamides, which evolved from a related series of HCV NS5b polymerase inhibitors, have been optimized to provide selective HIV integrase strand transfer inhibitors. Extensive SAR around the benzylamide moiety led to the identification of the p-fluorobenzylamide as optimal in the enzymatic assay. PMID- 17107800 TI - Leflunomide-induced peripheral neuropathy. AB - Two cases of leflunomide-induced peripheral neuropathy are described; in a 60 year-old woman with sero-negative polyarthralgia-myalgia syndrome and a 65-year old man with sero-negative rheumatoid arthritis, both treated with leflunomide at 20mg/day. Nerve conduction studies and electromyogram showed sensorimotor axonal neuropathy in both cases. An alternative cause for the axonal neuropathy was excluded by extensive investigations, including cerebrospinal fluid examination and nerve biopsy in the second patient. Both patients stabilized symptomatically and electrophysiologically upon cessation of leflunomide. It is possible that leflunomide-induced peripheral neuropathy has been under-reported and under recognized. PMID- 17107801 TI - Neurological sequelae of intrauterine warfarin exposure. AB - Warfarin embryopathy is a well-defined manifestation of intrauterine warfarin exposure. The embryopathy phenotype as it relates to the nervous system is broad and poorly recognised. We describe an adult with neurological sequelae of fetal warfarin exposure. We review previous cases with neurological sequelae and discuss the pathogenetic mechanism in light of recent research. PMID- 17107802 TI - Temporary alopecia after subarachnoid haemorrhage. AB - Primary endovascular intervention is increasingly the first choice of treatment for cerebral aneurysms, particularly for those with complex anatomy in the posterior circulation. However, their clinical management and follow-up continue to be predominantly in the hands of neurosurgeons. In this report, the development of alopecia following the coiling of posterior circulation aneurysms is described. The alopecia was transient and lasted for approximately 6 months, and occurred in the occipital and suboccipital regions of the scalp. This report aims to highlight this condition, which has not been previously reported in the neurosurgical literature. The potential hazards of irradiation should be borne in mind while carrying out complex endovascular procedures. The patient should be counselled and all necessary steps undertaken to limit radiation exposure. PMID- 17107803 TI - Utility of serum Campylobacter specific antibodies in determining prior Campylobacter infection in neurological disease. AB - Campylobacter jejuni has been implicated in the pathogenesis of Guillain-Barre syndrome (GBS); however, little information exists on the utility of Campylobacter serology in determining recent infection in the patient population. C. jejuni specific antibodies (CAs) were measured in 420 blood donors (controls), 99 patients with recent C. jejuni infection, 34 patients with central nervous system disorders (neurology controls), and 44 patients with peripheral nervous system (PNS) disorders: 18 with GBS, 12 with MND and 14 with chronic inflammatory neuropathies. Elevated CA titres consistent with recent C. jejuni infection were found in six of the 44 patients with PNS disorders (three with GBS, two with neuropathy, and one with MND, only one of whom had a history of recent C. jejuni infection), compared with two of 454 controls (p = 0.00001). Therefore, we conclude that CAs are often raised in patients with PNS disorders who do not have a history of recent C. jejuni gastroenteritis, so Campylobacter serology may be an unreliable marker of recent infection in this patient group. PMID- 17107804 TI - Optimal duration of conservative treatment for lumbar disc herniation depending on the type of herniation. AB - Six hundred and six clinical records of patients who were admitted to the Department of Orthopaedic Surgery, Suwa Red Cross Hospital, Nagano, Japan, due to lumbar disc herniation (LDH) between 1991 and 2002 were reviewed to determine the ideal conservative treatment period in relation to different types of LDH: contained and non-contained. Since 1997, we have prolonged in-hospital conservative treatment over a few weeks, which resulted in a decrease in the number of surgeries for non-contained LDH, especially of surgeries performed within 1 month. The number of surgeries for contained LDH did not decrease. The preoperative period of non-contained LDH was significantly shorter than that of contained LDH. These results indicate that for patients who have non-contained LDH, intensive conservative treatment may avoid a surgical procedure. For those patients who have contained LDH, conservative treatment should not be prolonged. We believe that the optimal duration of intensive conservative therapy should be less than 1 month. PMID- 17107805 TI - Synthesis, anti-tuberculosis activity, and 3D-QSAR study of 4-(adamantan-1-yl)-2 substituted quinolines. AB - Structural optimization of the previously identified 4-(adamantan-1-yl)-2 quinolinecarbohydrazide (AQCH, MIC=6.25 microg/mL, 99% inhibition, Mycobacterium tuberculosis H37Rv) has led to two series of 4-(adamantan-1-yl)-2-substituted quinolines (Series 1-2). All new derivatives were evaluated in vitro for antimycobacterial activities against drug-sensitive M. tuberculosis H37Rv strain. Several 4-adamantan-1-yl-quinoline-2-carboxylic acid N'-alkylhydrazides (Series 1) described herein showed promising inhibitory activity. In particular, analogs 7, 9, 20, and 21 displayed MIC of 3.125 microg/mL. Further investigation of AQCH by its reaction with various aliphatic, aromatic, and heteroaromatic aldehydes led to the synthesis of 4-adamantan-1-yl-quinoline-2-carboxylic acid alkylidene hydrazides (Series 2). Analogs 42-44 and 48 have produced promising antimycobacterial activities (99% inhibition) at 3.125 microg/mL against drug sensitive M. tuberculosis H37Rv strain. The most potent analog 35 of the series produced 99% inhibition at 1.00 microg/mL against drug-sensitive strain, and MIC of 3.125 microg/mL against isoniazid-resistant TB strain. To understand the relationship between structure and activity, a 3D-QSAR analysis has been carried out by three methods-comparative molecular field analysis (CoMFA), CoMFA with inclusion of a hydropathy field (HINT), and comparative molecular similarity indices analysis (CoMSIA). Several statistically significant CoMFA, CoMFA with HINT, and CoMSIA models were generated. Prediction of the activity of a test set of molecules was the best for the CoMFA model generated with database alignment. Based on the CoMFA contours, we have tried to explain the structure-activity relationships of the compounds reported herein. PMID- 17107806 TI - Linker-modified triamine-linked acridine dimers: synthesis and cytotoxicity properties in vitro and in vivo. AB - The preparation and cytotoxicity properties of a series of N(epsilon)-substituted triamine-linked acridine dimers are described. Most acridine dimer derivatives reveal highly potent in vitro cytotoxicity properties and DNA binding activity. Several acridine dimers were selected to study their action in vivo. These acridine dimers have demonstrated a narrow safe margin, as has adriamycin, but higher maximum tolerate dose (MTD) in comparison with that of adriamycin in ICR mice. The acridine dimers also demonstrated various anit-COLO 205 solid tumor activities in vivo. Compound 1 has shown the most potent solid tumor inhibition. PMID- 17107807 TI - Synthesis of novel potent hepatitis C virus NS3 protease inhibitors: discovery of 4-hydroxy-cyclopent-2-ene-1,2-dicarboxylic acid as a N-acyl-L-hydroxyproline bioisostere. AB - Potent tetrapeptidic inhibitors of the HCV NS3 protease have been developed incorporating 4-hydroxy-cyclopent-2-ene-1,2-dicarboxylic acid as a new N-acyl-l hydroxyproline mimic. The hydroxycyclopentene template was synthesized in eight steps from commercially available (syn)-tetrahydrophthalic anhydride. Three different amino acids were explored in the P1-position and in the P2-position the hydroxyl group of the cyclopentene template was substituted with 7-methoxy-2 phenyl-quinolin-4-ol. The P3/P4-positions were then optimized from a set of six amino acid derivatives. All inhibitors were evaluated in an in vitro assay using the full-length NS3 protease. Several potent inhibitors were identified, the most promising exhibiting a K(i) value of 1.1nM. PMID- 17107808 TI - Two simplified fluorescent staining techniques to observe infection structures of the oomycete Plasmopara viticola in grapevine leaf tissues. AB - Plasmopara viticola, the causal agent of grapevine downy mildew, is an obligate biotrophic oomycete that grows in the intercellular spaces of host tissues and develops haustoria in the cells. Histological observations are the most effective methods to visualize and quantify the development of the infection structures. We chose two staining techniques leading to high resolution and contrast between parasite structures and host-plant tissues with a minimum of sample preparation: Blankophor and KOH-aniline blue fluorescent stainings. Blankophor (50 ppm in water or 15% KOH) staining was used to study the zoospore encystement on the leaf surface after release from sporangia. The aniline blue dye (0.05% in 0.067 M K(2)HPO(4), pH 9-9.5, after hot KOH whitening) was used to observe the invasive structures inside host tissues that lead to the production of sporangiophores and infectious sporangia. We tested modifications of some parameters of the procedures to determine the most appropriate for high throughput analyses adapted to our pathosystem and equipment facilities. PMID- 17107809 TI - Mass attenuation coefficient of the Earth, Moon and Mars samples over 1 keV-100 GeV energy range. PMID- 17107810 TI - Linear transformation of EPR spectra as a method proposed for improving identification of paramagnetic species in ceramic. AB - In the present paper the paramagnetic centres in pottery generated after gamma irradiation were investigated. The computer resolution enhancement method (CREM) was used to increase the electron paramagnetic resonance (EPR) spectra resolution. The measurements were performed at room and liquid nitrogen temperatures. The paper shows the results of non-irradiated and gamma-irradiated pottery before and after resolution enhancement. Each EPR spectrum showed a multiple structure after the CREM procedure application. An aluminium hole centre [AlO(4)](0) has been ascribed to these lines. The g factors are: g(1)=2.0602, g(2)=2.0079, and g(3)=2.0019 and hyperfine splitting is A(1)=5.5G and A(2) approximately A(3)=6.3G. PMID- 17107811 TI - 2-Benzoxazolinone (BOA) induced oxidative stress, lipid peroxidation and changes in some antioxidant enzyme activities in mung bean (Phaseolus aureus). AB - 2-Benzoxazolinone (BOA), a well-known allelochemical with strong phytotoxicity, is a potential herbicidal candidate. The aim of the present study was to determine whether phytotoxicity of BOA is due to induction of oxidative stress caused by generation of reactive oxygen species (ROS) and the changes in levels of antioxidant enzymes induced in response to BOA. Effect of BOA was studied on electrolyte leakage, lipid peroxidation (LP), hydrogen peroxide (H(2)O(2)) generation, proline (PRO) accumulation, and activities of antioxidant enzymes superoxide dismutase (SOD, 1.15.1.1), ascorbate peroxidase (APX, 1.11.1.11), guaiacol peroxidase (GPX, 1.11.1.7), catalase (CAT, 1.11.1.6) and glutathione reductase (GR, 1.6.4.2) in Phaseolus aureus (mung bean). BOA significantly enhanced malondialdehyde (MDA) content, a product of LP, in both leaves and roots of mung bean. The amount of H(2)O(2), a product of oxidative stress, and endogenous PRO increased many-fold in response to BOA. Accumulation of PRO, MDA and H(2)O(2) indicates the cellular damage in the target tissue caused by ROS generated by BOA. In response to BOA, there was a significant increase in the activities of scavenging enzymes SOD, APX, GPX, CAT, and GR in root and leaf tissue of mung bean. At 5 mM BOA, GR activity in roots showed a nearly 22-fold increase over that in control. The present study concludes that BOA induces oxidative stress in mung bean through generation of ROS and upregulation of activities of various scavenging enzymes. PMID- 17107812 TI - Possible role of jasmonic acid in the regulation of floral induction, evocation and floral differentiation in Lemna minor L. AB - Jasmonic acid (JA) is implicated in a wide variety of developmental and physiological processes in plants. Here, we studied the effects of JA and the combination of JA and ethylenediamine-dio-hydroxyphenyl-acetic acid (EDDHA) on flowering in Lemna minor in axenical cultures. JA (0.475-47.5 nmol l(-1)) enhanced floral induction in L. minor under long-day (LD) conditions. Under the same conditions, at a concentration of 237.5 nmol l(-1), JA inhibited floral induction, and at a concentration of 475 nmol l(-1) it prevented floral induction. Under LD conditions with LD preculture, a combination of EDDHA (20,500 nmol l(-1)) and JA (47.5 nmol l(-1)) had a synergistic effect on the promotion of floral induction. Floral induction was enhanced to the greatest extent in experiments with LD precultures. Microscopic examination of microphotographs of histological sections showed that JA and, to an even greater extent, JA+EDDHA at optimal concentrations promote apical floral induction (evocation). Furthermore, JA, and to an even greater extent JA in combination with EDDHA in an optimal concentration, also promote flower differentiation, especially the development of stamens, as is evident from the microphotographs. The experimental results show that JA promotes floral induction in other species of Lemnaceae from various groups according to their photoperiodic response. The results support our hypothesis that, in addition to previously ascribed functions, JA may regulate floral induction, evocation and floral differentiation. Our hypothesis is supported also by the results obtained by quantitative determination of endogenous JA levels in L. minor at three growth stages. The levels of endogenous JA decreased from 389 ng JA g(-1) (fresh weight) of L. minor during the vegetative stage to 217 ng JA g(-1) during the evocation stage, and to 37.5 ng JA g(-1) during the flowering stage, which proves that JA is used for flowering. PMID- 17107813 TI - Eucalyptus gunnii CCR and CAD2 promoters are active in lignifying cells during primary and secondary xylem formation in Arabidopsis thaliana. AB - Cell-specific expression patterns of the Eucalyptus gunnii cinnamoyl coenzymeA reductase (EgCCR) and cinnamyl alcohol dehydrogenase (EgCAD2) promoters were analyzed by promoter-GUS histochemistry in the primary and secondary xylem tissues from floral stems and roots of Arabidopsis thaliana. Expression patterns indicated that the EgCCR and EgCAD2 genes were expressed in a coordinated manner in primary and secondary xylem tissues of the Arabidopsis floral stem and root. Both genes were expressed in all lignifying cells (vessel elements, xylem fibers and paratracheal parenchyma cells) of xylem tissues. The capacity for long-term monolignol production appeared to be related to the cell-specific developmental processes and biological roles of different cell types. Our results suggested that lignification of short-lived vessel elements was achieved by a two-step process involving (i) monolignol production by vessel elements prior to vessel programmed cell death and (ii) subsequent monolignol production by vessel associated living paratracheal parenchyma cells following vessel element cell death. EgCCR and EgCAD2 gene expression patterns suggested that the process of xylem cell lignification was similar in both primary and secondary xylem tissues in Arabidopsis floral stems and roots. PMID- 17107814 TI - Jasmonates induce Nod factor production by Bradyrhizobium japonicum. AB - Jasmonates are signaling molecules involved in induced systemic resistance, wounding and stress responses of plants. We have previously demonstrated that jasmonates can induce nod genes of Bradyrhizobium japonicum when measured by beta galactosidase activity. In order to test whether jasmonates can effectively induce the production and secretion of Nod factors (lipo-chitooligosaccharides, LCOs) from B. japonicum, we induced two B. japonicum strains, 532C and USDA3, with jasmonic acid (JA), methyl jasmonate (MeJA) and genistein (Ge). As genistein is well characterized as an inducer of nod genes it was used a positive control. The high-performance liquid chromatography (HPLC) profile of LCOs isolated following treatment with jasmonates or genistein showed that both JA and MeJA effectively induced nod genes and caused production of LCOs from bacterial cultures. JA and MeJA are more efficacious inducers of LCO production than genistein. Genistein plus JA or MeJA resulted in greater LCO production than either alone. A soybean root hair deformation assay showed that jasmonate induced LCOs were as effective as those induced by genistein. This is the first report that jasmonates induce Nod factor production by B. japonicum. This report establishes the role of jasmonates as a new class of signaling molecules in the Bradyrhizobium-soybean symbiosis. PMID- 17107815 TI - Nomenclatural status and identity of Gobius lagocephalus (Teleostei: Gobiidae). PMID- 17107816 TI - Phylogeography of the human mitochondrial L1c haplogroup: genetic signatures of the prehistory of Central Africa. AB - Interindividual variation of human mitochondrial DNA has been extensively studied over the last two decades, and its usefulness for reconstructing evolutionary relationships of extant populations has been proved. However, some mitochondrial lineages still need to be studied using a combination of larger and tailored datasets and increased level of resolution in order to shed light on their origin and on the processes underlying their present distribution. In this study, we analyze the phylogeny of the L1c haplogroup of human mitochondrial DNA using sequence data from hypervariable regions 1 and 2 obtained from 455 individuals (extracted from a total sampling of 2542 individuals) belonging to sub-Saharan African and African-American populations. We propose a substantial revision of L1c phylogeny, by introducing one new sub-haplogroup (L1c4), two new L1c1 clades (L1c1b and L1c1c), and by reassigning the previous L1c1a1 sequences to a clade which we termed L1c5. The new phylogeny encompasses distinct lineages with different evolutionary histories. In fact, based on population frequency, internal variation and mismatch distribution, we propose that L1c1b, L1c1c and L1c2 originated in Bantu ancestors, whereas L1c1a, L1c4 and L1c5 evolved among Western Pygmies. The population structure of L1c is not comparable to any known mitochondrial or, even, Y-chromosomal haplogroup, and challenges the current view that most of mtDNA variation in Pygmies might reflect admixture with Bantu or a persistence of plesiomorphic characters. In fact, the unique feature of the L1c is that it retains a signature of a phase common to the ancestors of the Bantu and Western Pygmies, while encompassing some specific sub-clades which can indicate their divergence. This allowed us to attempt a phylogenetically based assessment of the evolutionary relationships between the two groups. Taking into consideration estimates of the time to the most recent common ancestor of L1c and its clades together with archaeological and paleoclimatological evidence, we propose that the ancestors of Bantu and Western Pygmies separated between 60 and 30 kya. PMID- 17107817 TI - MgMfs1, a major facilitator superfamily transporter from the fungal wheat pathogen Mycosphaerella graminicola, is a strong protectant against natural toxic compounds and fungicides. AB - MgMfs1, a major facilitator superfamily (MFS) gene from the wheat pathogenic fungus Mycosphaerella graminicola, was identified in expressed sequence tag (EST) libraries. The encoded protein has high homology to members of the drug:H(+) antiporter efflux family of MFS transporters with 14 predicted transmembrane spanners (DHA14), implicated in mycotoxin secretion and multidrug resistance. Heterologous expression of MgMfs1 in a hypersensitive Saccharomyces cerevisiae strain resulted in a strong decrease in sensitivity of this organism to a broad range of unrelated synthetic and natural toxic compounds. The sensitivity of MgMfs1 disruption mutants of M. graminicola to most of these compounds was similar when compared to the wild-type but the sensitivity to strobilurin fungicides and the mycotoxin cercosporin was increased. Virulence of the disruption mutants on wheat seedlings was not affected. The results indicate that MgMfs1 is a true multidrug transporter that can function as a determinant of pathogen sensitivity and resistance to fungal toxins and fungicides. PMID- 17107818 TI - Impact of chronic pain on everyday physical activity. AB - Although patients with chronic pain are often considered to have reduced levels of everyday physical activity, data on their activity levels are scarce and inconclusive. Therefore, this study explored whether patients with chronic pain have reduced activity levels, as objectively measured with an activity monitor. The activity monitor is based on long-term ambulatory monitoring of signals from body-fixed accelerometers during everyday life, aimed at assessment of mobility related activities. Measurements with the monitor were performed during a weekday (24 h) in 18 patients with chronic pain and compared with measurements obtained from 18 gender and age matched healthy comparison subjects. The mean (SD) age of the patients was 44 (11) years, and the mean (SD) duration of their complaints was 8 (7) years. Compared with the healthy subjects, the duration of dynamic activities was not significantly reduced (p=0.10) in the patient group. Mean (SD) intensity of everyday physical activity was lower (p=0.03) in the patients than in the healthy comparison subjects (0.021 [0.006] g versus 0.026 [0.004] g), and patients spent more time lying down (47.0 [10.2]% versus 34.3 [5.6] %; p=0.000) and less time sitting (29.2 [8.9]% versus 36.4 [9.3]%; p=0.03) than the healthy comparison subjects. In spite of significant differences between patients and healthy comparison subjects for some aspects of the activity pattern (which may reflect pain behaviour), the impact of chronic pain on everyday physical activity was relatively small. PMID- 17107819 TI - Testosterone effects on avian basal metabolic rate and aerobic performance: facts and artefacts. AB - We examined the effects of cage size and testosterone (T) levels on basal and peak metabolic rates (BMR and PMR, respectively) and on pectoral and leg muscle masses of male house sparrows (Passer domesticus). Birds were housed either in small birdcages or in flight aviaries for at least 2 weeks prior to the initial metabolic evaluations. They were then implanted with either empty or T-filled silastic capsules and remeasured 5-6 weeks later. Birds treated with single T implants achieved breeding levels (4-6 ng/mL) and one group given double implants reached 10 ng/mL. There was no effect of T on BMR or PMR in any group studied, but there was an effect of caging. Caged birds showed significant reductions in PMR over the course of captivity, whereas PMR in aviary-housed birds were indistinguishable from their free-living counterparts. Testosterone treatment significantly increased leg muscle mass in caged birds, but had no effect on muscle mass in aviary-housed sparrows. We conclude that testosterone has no direct effect on sparrow metabolic rate or muscle mass, but may interact with cage conditions to produce indirect changes to these variables. PMID- 17107820 TI - Are the carotid bodies of the guinea-pig functional? AB - We have previously observed that the guinea-pig appears to have a relatively poor ventilatory (V (E)) response to hypoxia, compared to other mammals. Therefore, in this study, we questioned the ability of the carotid bodies (primary peripheral chemoreceptors) in the guinea-pig to detect hypoxia. The ventilatory responses to poikilocapnic hypoxia (8% O(2)), poikilooxic hypercapnia (8% CO(2)), hyperoxia (100% O(2)) and cyanide (NaCN - 200 mug/kg, i.v.) were assessed before and after carotid body denervation (CBD) in anaesthetized guinea-pigs. Although CBD attenuated the V (E) responses to hypercapnia and cyanide, it had no effect on normoxic breathing or the V (E) responses to hypoxia or hyperoxia. In a separate group of guinea-pigs, nerve activity was recorded from single or few-fibre preparations of the carotid sinus nerve (CSN). Basal chemoreceptor activity could not be detected from any of the nerve preparations. NaCN and hypercapnia consistently provoked an increase in neural activity. In contrast, hypoxia never clearly increased activity in any of the single or few-fibre preparations isolated from the CSN. In conclusion, although the carotid bodies of the guinea pig, like those of other mammals, are able to detect hypercapnia and histotoxic hypoxia and elicit a reflex increase in V (E), they are essentially hypoxia insensitive. The latter may explain, at least in part, the relatively poor V (E) response to hypoxia shown by the guinea-pig. PMID- 17107821 TI - Skeletal muscle IGF-I isoform expression in healthy women after isometric exercise. AB - CONTEXT: Little is known about the expression of skeletal muscle-specific isoforms of the insulin-like growth factor-I (IGF-I) gene in response to exercise in humans. Data for women are sparse and the influences of age and physical training status are unknown. Here we aimed to describe the expression of mRNA for the IGF-I isoforms in the quadriceps muscles of women at rest and after isometric exercise. OBJECTIVE: To categorize mRNA expression of the IGF-I splice variants IGF-IEa and IGF-IEc (MGF) in healthy women, determine whether isometric exercise stimulates expression, and to determine the duration of the response. DESIGN: Observational study. SETTING: Clinical research facility. PARTICIPANTS: Six healthy women (median age 38 years, range 25-43 years). INTERVENTION: Single bout of maximal isometric knee extension undertaken as 20 sets of 6 contractions. Muscle samples from the lateral mass of the quadriceps were obtained by needle biopsy at baseline, 2.5h and 50h after exercise. MAIN OUTCOME MEASURES: Quantification of mRNAs for IGF-IEa and MGF. RESULTS: mRNA for the IGF-IEa transcript was significantly elevated by 50h post-exercise compared to baseline (p=0.005) and mRNA for MGF was significantly elevated by 2.5h (p=0.026). There were no statistically significant differences between measurements at 2.5h and 50h for either transcript. CONCLUSIONS: We have shown that the upregulation of the muscle specific IGF-IEa and MGF isoforms occurs within 2.5h of a single bout of isometric exercise in women. The upregulation persists for at least 2 days after exercise. We have also demonstrated a potentially safe and effective way of studying the responsiveness of these isoforms to resistance exercise in future studies of older and/or frail individuals. PMID- 17107822 TI - Writer's cramp: restoration of striatal D2-binding after successful biofeedback based sensorimotor training. AB - INTRODUCTION: Previous studies of writer's cramp have detected cerebral sensorimotor abnormalities in this disorder and, more specifically, a reduced striatal D2-binding as assessed by [(123)I]IBZM SPECT. However, empirical data were lacking about the influence of effective biofeedback-based sensorimotor training on D2 receptor binding. METHODS: To determine whether there is a restoration of D2-binding after successful sensorimotor treatment, pre- and posttreatment SPECTs were compared in five patients with writer's cramp and correlated with improvement in handwriting. RESULTS: After treatment, the clinical and electromyographic picture appeared substantially improved connected with a significant increase in D2-binding to nearly normal levels similar to normative data in age/sex-matched healthy subjects. CONCLUSION: The current study supported the view that writer's cramp results from a plastic adaptation of a rectifiable nigrostriatal dopaminergic system and that effective sensorimotor training leads to increased efficacy of striatal dopaminergic transmission. PMID- 17107823 TI - What's happening - Scotblood 2006 advances in immunetolerance, information technology, microarrays and pathogen inactivation in transfusion medicine. AB - This commentary on Scotblood meeting aimed to provide a highlight on four areas of new development in blood transfusion medicine, based on the abstracts provided by the invited speakers. Dr. Jerard Seghatchian would like to express his sincere thanks to Prof. Robin Fraser, Chairman of the Scotblood Organising Committee, and Dr. Hagop Bessos, chairman of the Scotblood Programme Sub-Committee, for inviting him to attend this event and providing him with the essential material for this report. PMID- 17107825 TI - Factors associated with loss to follow-up in a large tuberculosis treatment trial (TBTC Study 22). AB - INTRODUCTION: Loss to follow-up in clinical trials compromises achievement of study goals. We evaluated factors associated with loss to follow-up after completion of treatment phase in a large tuberculosis treatment trial (TBTC/USPHS Study 22) in the U.S. and Canada. METHODS: Patients who were lost to follow-up were compared to those who reached a study end-point or successfully completed follow-up. A generalized estimating equation model was used to combine patient specific and site-specific factors. RESULTS: Of 1075 patients enrolled, 965 (89.8%) reached a study end-point, died, or completed the 2 year post-treatment follow-up phase, and 110 (10.2%) did not. Multivariate analysis showed the following factors to be independently associated with loss to follow-up: birth outside USA/Canada (OR 2.07, 95% CI 1.25-3.40, p=0.005), history of homelessness (OR 1.94, 95% CI 1.00-3.80, p=0.05), enrollment at a health department (OR 2.71, 95% CI 1.27-5.79, p=0.010), and use of any kind of incentive (cash/cash equivalent) during treatment phase (OR 3.04, 95% CI 1.73-5.33 p=0.0001). CONCLUSIONS: Cultural or linguistic factors and lack of stable housing contribute to loss to follow-up. Attention to these factors could improve long-term retention in clinical trials. Enrollment at a health department and use of incentives during treatment phase may be markers for other factors leading to loss to follow-up. PMID- 17107826 TI - Direct electrochemistry of glucose oxidase immobilized on a hexagonal mesoporous silica-MCM-41 matrix. AB - The direct electrochemistry of glucose oxidase (GOD) immobilized on a hexagonal mesoporous silica modified glassy carbon electrode was investigated. The adsorbed GOD displayed a pair of redox peaks with a formal potential of -417 mV in 0.1 M pH 6.1 phosphate buffer solution (PBS). The response showed a diffusion controlled electrode process with a two-electron transfer coupled with a two proton transfer reaction process. GOD immobilized on a hexagonal mesoporous silica retained its bioactivity and stability. In addition, the immobilized GOD could electrocatalyze the oxidation of glucose to gluconlactone by taking ferrocene monocarboxylic acid (FMCA) as a mediator in N(2) saturated solutions, indicating that the electrode may have the potential application in biosensors to analyze glucose. The sensor could exclude the interference of commonly coexisted uric acid, p-acetaminophenol and ascorbic acid and diagnose diabetes very fast and sensitively. This work demonstrated that the mesoporous silica provided a novel matrix for protein immobilization and the construction of biosensors. PMID- 17107827 TI - Plastic reference electrodes and plastic potentiometric cells with dispersion cast poly(3,4-ethylenedioxythiophene) and poly(vinyl chloride) based membranes. AB - A simple procedure of preparing low cost, planar and disposable reference electrodes for potentiometric applications is presented. This method is essentially the same as used for obtaining all-plastic ion-selective electrodes and thus promising for simple fabrication of complete cells. Commercially available aqueous dispersion of poly(3,4-ethylenedioxythiophene) doped by poly(4 styrenesulfonate) ions (PEDOT-PSS, Baytron P) is simply cast on a non-conducting plastic support (transparent foil for laser printers). This layer is covered by a non-selective poly(vinyl chloride) based membrane containing solid AgCl and KCl, added to obtain a stable potential. The conducting polymer layer plays a double role, of electrical contact and ion-to-electron transducer, enhancing the potential stability. The reference electrodes obtained exhibit independence of the kind and concentration of electrolyte applied as well as very low sensitivity to interferences: redox reactants and H+ ions; they are also characterized by both potential stability and low polarisability, sufficient for potentiometric applications. Cells of plastic electrodes (indicator and reference ones) are tested using an arrangement with Pb2+ or Ca2+ selective sensors. Potentiometric characteristic of such cells is satisfactory, well comparable with that using a classical electrode arrangement. PMID- 17107828 TI - Focusing on global environmental health. PMID- 17107829 TI - Hormesis: a new religion? PMID- 17107831 TI - Suggested corrections to the Farm Family Exposure Study. PMID- 17107833 TI - Risk assessment and epidemiologic evidence in environmental health science. PMID- 17107834 TI - Chemical exposures. Will DEA findings wash? PMID- 17107837 TI - Global warming. Clinton climate initiative heats up. PMID- 17107836 TI - Reproductive toxicity. New take on perchlorate effects. PMID- 17107840 TI - The feed factor: estrogenic variability in lab animal diets. PMID- 17107842 TI - Fertile grounds of inquiry: environmental effects on human reproduction. PMID- 17107843 TI - Setting a baseline for biomonitoring. PMID- 17107844 TI - Fe-TAML: catalyst for cleanup. PMID- 17107845 TI - The importance of hormesis to public health. AB - BACKGROUND: Hormesis is a specific type of nonmonotonic dose response whose occurrence has been documented across a broad range of biological models, diverse types of exposure, and a variety of outcomes. The effects that occur at various points along this curve can be interpreted as beneficial or detrimental, depending on the biological or ecologic context in which they occur. OBJECTIVE: Because hormesis appears to be a relatively common phenomenon that has not yet been incorporated into regulatory practice, the objective of this commentary is to explore some of its more obvious public health and risk assessment implications, with particular reference to issues raised recently within this journal by other authors. DISCUSSION: Hormesis appears to be more common than dose-response curves that are currently used in the risk assessment process [e.g., linear no-threshold (LNT)]. Although a number of mechanisms have been identified that explain many hormetic dose-response relationships, better understanding of this phenomenon will likely lead to different strategies not only for the prevention and treatment of disease but also for the promotion of improved public health as it relates to both specific and more holistic health outcomes. CONCLUSIONS: We believe that ignoring hormesis is poor policy because it ignores knowledge that could be used to improve public health. PMID- 17107847 TI - Decreased serum free testosterone in workers exposed to high levels of di-n-butyl phthalate (DBP) and di-2-ethylhexyl phthalate (DEHP): a cross-sectional study in China. AB - BACKGROUND: Observations of adverse developmental and reproductive effects in laboratory animals and wildlife have fueled increasing public concern regarding the potential for various chemicals to impair human fertility. OBJECTIVE: Our objective in this study was to assess the effect of occupational exposure to high levels of phthalate esters on the balance of gonadotropin and gonadal hormones including luteinizing hormone, follicle-stimulating hormone, free testosterone (fT), and estradiol. METHODS: We examined urine and blood samples of 74 male workers at a factory producing unfoamed polyvinyl chloride flooring exposed to di n-butyl phthalate (DBP) and di-2-ethylhexyl phthalate (DEHP) and compared them with samples from 63 male workers from a construction company, group matched for age and smoking status. RESULTS: Compared to the unexposed workers, the exposed workers had substantially and significantly elevated concentrations of mono-n butyl phthalate (MBP; 644.3 vs. 129.6 microg/g creatinine, p < 0.001) and mono-2 ethylhexyl phthalate (MEHP; 565.7 vs. 5.7 microg/g creatinine, p < 0.001). fT was significantly lower (8.4 vs. 9.7 microg/g creatinine, p = 0.019) in exposed workers than in unexposed workers. fT was negatively correlated to MBP (r = 0.25, p = 0.03) and MEHP (r = -0.19, p = 0.095) in the exposed worker group. Regression analyses revealed that fT decreases significantly with increasing total phthalate ester score (the sum of quartiles of MBP and MEHP; r = -0.26, p = 0.002). CONCLUSION: We observed a modest and significant reduction of serum fT in workers with higher levels of urinary MBP and MEHP compared with unexposed workers. PMID- 17107846 TI - Exposures to airborne particulate matter and adverse perinatal outcomes: a biologically plausible mechanistic framework for exploring potential effect modification by nutrition. AB - OBJECTIVES: The specific objectives are threefold: to describe the biologically plausible mechanistic pathways by which exposure to particulate matter (PM) may lead to the adverse perinatal outcomes of low birth weight (LBW), intrauterine growth retardation (IUGR), and preterm delivery (PTD); review the evidence showing that nutrition affects the biologic pathways; and explain the mechanisms by which nutrition may modify the impact of PM exposure on perinatal outcomes. METHODS: We propose an interdisciplinary conceptual framework that brings together maternal and infant nutrition, air pollution exposure assessment, and cardiopulmonary and perinatal epidemiology. Five possible albeit not exclusive biologic mechanisms have been put forth in the emerging environmental sciences literature and provide corollaries for the proposed framework. CONCLUSIONS: Protecting the environmental health of mothers and infants remains a top global priority. The existing literature indicates that the effects of PM on LBW, PTD, and IUGR may manifest through the cardiovascular mechanisms of oxidative stress, inflammation, coagulation, endothelial function, and hemodynamic responses. PM exposure studies relating mechanistic pathways to perinatal outcomes should consider the likelihood that biologic responses and adverse birth outcomes may be derived from both PM and non-PM sources (e.g., nutrition). In the concluding section, we present strategies for empirically testing the proposed model and developing future research efforts. PMID- 17107848 TI - Serum dioxin, testosterone, and subsequent risk of benign prostatic hyperplasia: a prospective cohort study of Air Force veterans. AB - BACKGROUND: Operation Ranch Hand veterans were involved in spraying herbicides, including Agent Orange, during the Vietnam War in 1962-1971; Agent Orange was contaminated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). It has been hypothesized that dioxins may be partially responsible for an increase of male reproductive tract disorders such as testicular cancer, cryptorchidism, and hypospadias. OBJECTIVES: In this study, our objective was to assess the effect of serum TCDD concentration on the risk of development of benign prostatic hyperplasia (BPH) and on serum testosterone levels. METHODS: This study was a longitudinal, prospective cohort study made up of U.S. Air Force veterans involved in Operation Ranch Hand. Other Air Force veterans who did not spray herbicides were included as comparisons. BPH was determined by medical record review and by medical examinations conducted during the study. Data were available for 971 Ranch Hand and 1,266 comparison veterans. We investigated the relationship between BPH and serum TCDD level using the Cox proportional hazards models adjusted for testosterone levels, body mass index (BMI), and the percentage change in BMI per year. RESULTS: In univariate and multivariate analyses, the risk of BPH decreased with increasing serum TCDD in the comparison group. The multivariate risk ratio for BPH in the comparison group was 0.84 (95% confidence interval, 0.73-0.98). Excluding men with prostate cancer, inflammatory or other prostatic diseases did not substantially alter the association. Serum testosterone levels were inversely associated with serum TCDD levels in both Ranch Hand and comparison groups. CONCLUSIONS: TCDD exposure at general population levels is associated with a decreasing risk of BPH with higher exposure levels. TCDD exposure is also negatively associated with serum testosterone levels. PMID- 17107849 TI - Expression of xenobiotic metabolizing enzymes in different lung compartments of smokers and nonsmokers. AB - BACKGROUND: Cytochrome P450 monooxygenases (CYP) play an important role in the defense against inhaled toxicants, and expression of CYP enzymes may differ among various lung cells and tissue compartments. METHODS: We studied the effects of tobacco smoke in volunteers and investigated gene expression of 19 CYPs and 3 flavin-containing monooxygenases, as well as isoforms of glutathione S transferases (GST) and uridine diphosphate glucuronosyltransferases (UGT) and the microsomal epoxide hydrolase (EPHX1) in bronchoalveolar lavage cells and bronchial biopsies derived from smokers (n = 8) and nonsmokers (n = 10). We also investigated gene expression of nuclear transcription factors known to be involved in the regulation of xenobiotic metabolism enzymes. RESULTS: Gene expression of CYP1A1, CYP1B1, CYP2S1, GSTP1, and EPHX1 was induced in bronchoalveolar lavage cells of smokers, whereas expression of CYP2B6/7, CYP3A5, and UGT2A1 was repressed. In bronchial biopsies of smokers, CYP1A1, CYP1B1, CYP2C9, GSTP1, and GSTA2 were induced, but CYP2J2 and EPHX1 were repressed. Induction of CYP1A1 and CYP1B1 transcript abundance resulted in increased activity of the coded enzyme. Finally, expression of the liver X receptor and the glucocorticoid receptor was significantly up-regulated in bronchoalveolar lavage cells of smokers. CONCLUSIONS: We found gene expression of pulmonary xenobiotic metabolizing enzymes and certain key transcription factors to be regulated in bronchoalveolar lavage cells and bronchial biopsies of smokers. The observed changes demonstrate tissue specificity in xenobiotic metabolism, with likely implications for the metabolic activation of procarcinogens to ultimate carcinogens of tobacco smoke. PMID- 17107850 TI - Cardiovascular effects of nickel in ambient air. AB - BACKGROUND: Fine particulate matter (FPM) in ambient air causes premature mortality due to cardiac disease in susceptible populations. OBJECTIVE: Our objective in this study was to determine the most influential FPM components. METHODS: A mouse model of atherosclerosis (ApoE-/-) was exposed to either filtered air or concentrated FPM (CAPs) in Tuxedo, New York (85 microg/m3 average, 6 hr/day, 5 days/week, for 6 months), and the FPM elemental composition was determined for each day. We also examined associations between PM components and mortality for two population studies: National Mortality and Morbidity Air Pollution Study (NMMAPS) and Hong Kong. RESULTS: For the CAPs-exposed mice, the average of nickel was 43 ng/m3, but on 14 days, there were Ni peaks at approximately 175 ng/m3 and unusually low FPM and vanadium. For those days, back trajectory analyses identified a remote Ni point source. Electrocardiographic measurements on CAPs-exposed and sham-exposed mice showed Ni to be significantly associated with acute changes in heart rate and its variability. In NMMAPS, daily mortality rates in the 60 cities with recent speciation data were significantly associated with average Ni and V, but not with other measured species. Also, the Hong Kong sulfur intervention produced sharp drops in sulfur dioxide, Ni, and V, but not other components, corresponding to the intervention-related reduction in cardiovascular and pulmonary mortality. CONCLUSIONS: Known biological mechanisms cannot account for the significant associations between Ni with the acute cardiac function changes in the mice or with cardiovascular mortality in people at low ambient air concentrations; therefore, further research is needed. PMID- 17107851 TI - Serum dioxin-like activity is associated with reproductive parameters in young men from the general Flemish population. AB - BACKGROUND: 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and some related environmental contaminants are aryl hydrocarbon receptor (AhR) ligands that exert reproductive and developmental toxicity in laboratory animals. In humans, fertility-related effects are less documented. OBJECTIVE: The aim of this study was to investigate the relationship between dioxin-like biological activity in serum and parameters of reproductive status in men from the general population 5 months after a polychlorinated biphenyl and dioxin food-contamination episode in Belgium. DESIGN: In the framework of the cross-sectional Flemish Environment and Health Study (FLEHS), we recruited 101 men 20-40 years of age and evaluated sperm parameters, measured sex hormones, and gathered information on a number of lifestyle factors. In addition, we determined the AhR-mediated enzymatic response elicited by individual serum samples and expressed it as TCDD equivalent concentrations (CALUX-TEQs) using an established transactivation assay. RESULTS: Age (p = 0.04) and the frequency of fish (p = 0.02) and egg (p = 0.001) consumption were independent positive determinants of serum dioxin-like activity. After correcting for possible confounders, we found that a 2-fold increase in CALUX-TEQ > 16 pg/L was associated with a 7.1% and 6.8% (both p = 0.04) decrease in total and free testosterone, respectively. We also observed a more pronounced drop in semen volume of 16.0% (p = 0.03), whereas sperm concentration rose by 25.2% (p = 0.07). No relationship was found with total sperm count or sperm morphology. CONCLUSIONS: These data suggest an interaction of dioxin-like compounds with the secretory function of the seminal vesicles or prostate, possibly indirectly through an effect on testosterone secretion, at levels not affecting spermatogenesis as such. PMID- 17107852 TI - Molecular epidemiologic evidence for diabetogenic effects of dioxin exposure in U.S. Air force veterans of the Vietnam war. AB - BACKGROUND: One of the outcomes positively associated with dioxin exposure in humans is type 2 diabetes. OBJECTIVES: This study was conducted in order to find the molecular biological evidence for the diabetogenic action of dioxin in adipose samples from Vietnam veterans. METHODS: We obtained 313 adipose tissue samples both from Vietnam veterans who were exposed to dioxin (Operation Ranch Hand) and from comparison veterans who served in Southeast Asia with no record of dioxin exposure. We conducted quantitative reverse-transcribed polymerase chain reaction studies on selected marker mRNAs from these samples. RESULTS: We found the most sensitive and reliable molecular indicator of dioxin-induced diabetes to be the ratio of mRNA of glucose transporter 4 (GLUT4) and nuclear transcription factor kappa B (NFkappaB), a marker of inflammation. This ratio showed significant correlations to serum dioxin residues and to fasting glucose among those in the Ranch Hand group and, surprisingly, even in the comparison group, who have low levels of dioxin comparable to the general public. Such a correlation in the comparison group was particularly significant among those with known risk factors such as obesity and family history of diabetes. CONCLUSIONS: These results show that the GLUT4:NFkappaB ratio is a reliable marker for the diabetogenic action of dioxin, particularly at very low exposure levels that are not much higher than those found in the general public, implying a need to address current exposure levels. PMID- 17107853 TI - An intervention to reduce residential insecticide exposure during pregnancy among an inner-city cohort. AB - BACKGROUND: We previously reported widespread insecticide exposure during pregnancy among inner-city women from New York City. Here we report on a pilot intervention using integrated pest management (IPM) to reduce pest infestations and residential insecticide exposures among pregnant New York City African American and Latina women (25 intervention and 27 control homes). METHODS: The IPM consisted of professional cleaning, sealing of pest entry points, application of low-toxicity pesticides, and education. Cockroach infestation levels and 2 week integrated indoor air samples were collected at baseline and one month postintervention. The insecticides detected in the indoor air samples were also measured in maternal and umbilical cord blood collected at delivery. RESULTS: Cockroach infestations decreased significantly (p = 0.016) after the intervention among intervention cases but not control households. Among the intervention group, levels of piperonyl butoxide (a pyrethroid synergist) were significantly lower in indoor air samples after the intervention (p = 0.016). Insecticides were detected in maternal blood samples collected at delivery from controls but not from the intervention group. The difference was significant for trans-permethrin (p = 0.008) and of borderline significance (p = 0.1) for cis-permethrin and 2 isopropoxyphenol (a propoxur metabolite). CONCLUSION: To our knowledge, this is the first study to use biologic dosimeters of prenatal pesticide exposure for assessing effectiveness of IPM. These pilot data suggest that IPM is an effective strategy for reducing pest infestation levels and the internal dose of insecticides during pregnancy. PMID- 17107854 TI - Does particulate matter modify the association between temperature and cardiorespiratory diseases? AB - BACKGROUND: A number of studies have shown that both temperature and air pollution are associated with health outcomes. In assessing air pollution effects, temperature is usually considered a confounder. However, only a few recent studies considered air pollution as confounders while assessing temperature effects. Few studies are available on whether or not air pollution modifies the temperature-disease relationship. METHODS: In this study, we used three parallel Poisson generalized additive models to examine whether particulate matter < 10 mum in aerodynamic diameter (PM10) modified the effects of minimum temperature on cardiorespiratory morbidity and mortality in Brisbane, Australia. RESULTS: Results show that PM10 statistically significantly modified the effects of temperature on respiratory and cardiovascular hospital admissions, all nonexternal-cause mortality, and cardiovascular mortality at different lags. The enhanced adverse temperature effects were found at higher levels of PM10, but no clear evidence emerged for interactive effects on respiratory and cardiovascular emergency visits. Three parallel models produced similar results, which strengthened the validity of findings. CONCLUSION: We conclude that it is important to evaluate the modification role of air pollution in the assessment of temperature-related health impacts. PMID- 17107855 TI - Distribution of nanoparticles in the see-through medaka (Oryzias latipes). AB - OBJECTIVE: Because the environmental fate of manufactured nanoparticles is considered an emerging environmental concern, I used water-suspended fluorescent nanoparticles (solid latex solution) to investigate the distribution of nanoparticles in the eggs and bodies of see-through medaka (Oryzias latipes). RESULTS: Particles 39.4-42,000 nm in diameter were adsorbed to the chorion of medaka eggs and accumulated in the oil droplets; 474-nm particles had the highest bioavailability to eggs. Particles 39.4 nm in diameter shifted into the yolk and gallbladder during embryonic development. Adult medaka accumulated 39.4-nm nanoparticles mainly in the gills and intestine when exposed to a 10-mg/L nanoparticle solution. Nanoparticles were also detected in the brain, testis, liver, and blood. Concentrations of nanoparticles in the blood of male and female medaka were 16.5 and 10.5 ng/mg blood protein, respectively. These results suggest that nanoparticles are capable of penetrating the blood-brain barrier and that they eventually reach the brain. Salinity-dependent acute toxicity was observed in medaka eggs exposed for 24 hr to nanoparticles. CONCLUSION: The bioavailability and toxicity of nanoparticles depend on environmental factors and multiple physicochemical properties. Further studies on the toxic effects of nanoparticles used in commercial products and their environmental relevance, are necessary to define the risks and benefit of nanomaterial applications. PMID- 17107856 TI - Genotoxicants target distinct molecular networks in neonatal neurons. AB - BACKGROUND: Exposure of the brain to environmental agents during critical periods of neuronal development is considered a key factor underlying many neurologic disorders. OBJECTIVES: In this study we examined the influence of genotoxicants on cerebellar function during early development by measuring global gene expression changes. METHODS: We measured global gene expression in immature cerebellar neurons (i.e., granule cells) after treatment with two distinct alkylating agents, methylazoxymethanol (MAM) and nitrogen mustard (HN2). Granule cell cultures were treated for 24 hr with MAM (10-1,000 microM) or HN2 (0.1-20 microM) and examined for cell viability, DNA damage, and markers of apoptosis. RESULTS: Neuronal viability was significantly reduced (p < 0.01) at concentrations > 500 microM for MAM and > 1.0 microM for HN2; this correlated with an increase in both DNA damage and markers of apoptosis. Neuronal cultures treated with sublethal concentrations of MAM (100 microM) or HN2 (1.0 microM) were then examined for gene expression using large-scale mouse cDNA microarrays (27,648). Gene expression results revealed that a) global gene expression was predominantly up-regulated by both genotoxicants; b) the number of down-regulated genes was approximately 3-fold greater for HN2 than for MAM; and c) distinct classes of molecules were influenced by MAM (i.e, neuronal differentiation, the stress and immune response, and signal transduction) and HN2 (i.e, protein synthesis and apoptosis). CONCLUSIONS: These studies demonstrate that individual genotoxicants induce distinct gene expression signatures. Further study of these molecular networks may explain the variable response of the developing brain to different types of environmental genotoxicants. PMID- 17107857 TI - Grand rounds: outbreak of hematologic abnormalities in a community of people exposed to leakage of fire extinguisher gas. AB - CONTEXT: Although there are ample data on the respiratory effects of exposure to fire extinguisher gas, the potential hematologic effects have not been fully documented. We conducted this study to determine the possible etiologic agent(s) for a decrease in red blood cells among community residents in Taipei, Taiwan, after they were exposed to leakage of mixed fire extinguishants containing bromotrifluoromethane (CF3Br, Halon 1301), bromochlorodifluoromethane (CF2BrCl, Halon 1211), and dichlorodifluoromethane (CCl2F2, CFC-12). CASE PRESENTATION: We studied 117 exposed residents who came into one hospital for physical examinations. We also selected age- and sex-matched referents for comparison from residents who came to the same hospital for health examinations. Nine months after the exposure to mixed fire extinguishants, 91 of the exposed residents came back for a second physical examination. In the first examination of the exposed residents, we found a significant reduction in red blood cell count and hemoglobin and a relationship between dose and response. DISCUSSION: After excluding iron-deficiency anemia, thalassemia, and other possible agents, we suspected that the hematologic effects might have resulted from pyrolytic products of CFC-12 and Halon 1211, which may contain phosgene, among other products. RELEVANCE TO CLINICAL PRACTICE: The acute transient hematologic effects observed in the exposed residents were associated with the incident of leakage of mixed fire-extinguisher gases and were most likely caused by a small amount of pyrolytic products, probably phosgene. Nine months after the exposure, we found a significant improvement in the abnormalities without any specific treatment. PMID- 17107858 TI - Low-level lead exposure, metabolic syndrome, and heart rate variability: the VA Normative Aging Study. AB - BACKGROUND: Altered heart rate variability (HRV), a marker of poor cardiac autonomic function, has been associated with sudden cardiac death and heart failure. OBJECTIVE: We examined the association of low-level lead exposure measured in bone by K-X-ray fluorescence with alterations in HRV, and whether metabolic syndrome (MetS) or its individual components modify those associations. METHODS: HRV measures [power in high-frequency (HFnorm) and low-frequency (LFnorm) in normalized units, and LF/HF] were taken among 413 elderly men from the Normative Aging Study. MetS was defined as subjects having three or more of the following criteria: abdominal obesity, hypertriglyceridemia, low high-density lipoprotein, high blood pressure, and high fasting glucose. RESULTS: Of the subjects, 32% were identified as having MetS. Inverse but nonstatistically significant associations of both tibia and patella lead levels with HFnorm and nonstatistically significant positive relations with LFnorm and LF/HF were found in the entire cohort. There was a graded, statistically significant reduction in HFnorm and increases in LFnorm and LF/HF in association with an increase in patella lead as the number of metabolic abnormalities increased. We also observed that higher patella lead was consistently associated with lower HFnorm and higher LFnorm and LF/HF among subjects with MetS or its individual components. No statistically significant interaction between MetS and tibia lead was observed. CONCLUSION: The results suggest that elderly men with MetS were more susceptible to autonomic dysfunction in association with chronic lead exposure as measured in patella. The modification by MetS is consistent with a role for oxidative stress in lead toxicity on the cardiovascular system. PMID- 17107859 TI - Changes induced by exposure of the human lung to glass fiber-reinforced plastic. AB - The inhalation of glass dusts mixed in resin, generally known as glass fiber reinforced plastic (GRP), represents a little-studied occupational hazard. The few studies performed have highlighted nonspecific lung disorders in animals and in humans. In the present study we evaluated the alteration of the respiratory system and the pathogenic mechanisms causing the changes in a group of working men employed in different GRP processing operations and exposed to production dusts. The study was conducted on a sample of 29 male subjects whose mean age was 37 years and mean length of service 11 years. All of the subjects were submitted to a clinical check-up, basic tests, and bronchoalveolar lavage (BAL); microscopic studies and biochemical analysis were performed on the BAL fluid. Tests of respiratory function showed a large number of obstructive syndromes; scanning electron microscopy highlighted qualitative and quantitative alterations of the alveolar macrophages; and transmission electron microscopy revealed the presence of electron-dense cytoplasmatic inclusions indicating intense and active phlogosis (external inflammation). Biochemical analyses highlighted an increase in protein content associated with alterations of the lung oxidant/antioxidant homeostasis. Inhalation of GRP, independent of environmental concentration, causes alterations of the cellular and humoral components of pulmonary interstitium; these alterations are identified microscopically as acute alveolitis. PMID- 17107860 TI - Fetal lead exposure at each stage of pregnancy as a predictor of infant mental development. AB - BACKGROUND: The impact of prenatal lead exposure on neurodevelopment remains unclear in terms of consistency, the trimester of greatest vulnerability, and the best method for estimating fetal lead exposure. OBJECTIVE: We studied prenatal lead exposure's impact on neurodevelopment using repeated measures of fetal dose as reflected by maternal whole blood and plasma lead levels. METHODS: We measured lead in maternal plasma and whole blood during each trimester in 146 pregnant women in Mexico City. We then measured umbilical cord blood lead at delivery and, when offspring were 12 and 24 months of age, measured blood lead and administered the Bayley Scales of Infant Development. We used multivariate regression, adjusting for covariates and 24-month blood lead, to compare the impacts of our pregnancy measures of fetal lead dose. RESULTS: Maternal lead levels were moderately high with a first-trimester blood lead mean (+/- SD) value of 7.1 +/- 5.1 microg/dL and 14% of values >or=10 microg/dL. Both maternal plasma and whole blood lead during the first trimester (but not in the second or third trimester) were significant predictors (p < 0.05) of poorer Mental Development Index (MDI) scores. In models combining all three trimester measures and using standardized coefficients, the effect of first-trimester maternal plasma lead was somewhat greater than the effect of first-trimester maternal whole blood lead and substantially greater than the effects of second- or third-trimester plasma lead, and values averaged over all three trimesters. A 1-SD change in first-trimester plasma lead was associated with a reduction in MDI score of 3.5 points. Postnatal blood lead levels in the offspring were less strongly correlated with MDI scores. CONCLUSIONS: Fetal lead exposure has an adverse effect on neurodevelopment, with an effect that may be most pronounced during the first trimester and best captured by measuring lead in either maternal plasma or whole blood. PMID- 17107861 TI - Personal and ambient air pollution is associated with increased exhaled nitric oxide in children with asthma. AB - BACKGROUND: Research has shown associations between pediatric asthma outcomes and airborne particulate matter (PM). The importance of particle components remains to be determined. METHODS: We followed a panel of 45 schoolchildren with persistent asthma living in Southern California. Subjects were monitored over 10 days with offline fractional exhaled nitric oxide (FeNO), a biomarker of airway inflammation. Personal active sampler exposures included continuous particulate matter < 2.5 microm in aerodynamic diameter (PM2.5), 24-hr PM2.5 elemental and organic carbon (EC, OC), and 24-hr nitrogen dioxide. Ambient exposures included PM2.5, PM2.5 EC and OC, and NO2. Data were analyzed with mixed models controlling for personal temperature, humidity and 10-day period. RESULTS: The strongest positive associations were between FeNO and 2-day average pollutant concentrations. Per interquartile range pollutant increase, these were: for 24 microg/m3 personal PM2.5, 1.1 ppb FeNO [95% confidence interval (CI), 0.1-1.9]; for 0.6 microg/m3 personal EC, 0.7 ppb FeNO (95% CI, 0.3-1.1); for 17 ppb personal NO2, 1.6 ppb FeNO (95% CI, 0.4-2.8). Larger associations were found for ambient EC and smaller associations for ambient NO2. Ambient PM2.5 and personal and ambient OC were significant only in subjects taking inhaled corticosteroids (ICS) alone. Subjects taking both ICS and antileukotrienes showed no significant associations. Distributed lag models showed personal PM2.5 in the preceding 5 hr was associated with FeNO. In two-pollutant models, the most robust associations were for personal and ambient EC and NO2, and for personal but not ambient PM2.5. CONCLUSION: PM associations with airway inflammation in asthmatics may be missed using ambient particle mass, which may not sufficiently represent causal pollutant components from fossil fuel combustion. PMID- 17107862 TI - International studies of prenatal exposure to polycyclic aromatic hydrocarbons and fetal growth. AB - OBJECTIVES: Polycyclic aromatic hydrocarbons (PAHs) are ubiquitously distributed human mutagens and carcinogens. However, lack of adequate air monitoring data has limited understanding of the effects of airborne PAHs on fetal growth. To address this gap in knowledge, we examined the association between prenatal exposure to airborne PAHs and birth weight, birth length, and birth head circumference, respectively, in Krakow, Poland, and New York City (NYC). METHODS: The parallel prospective cohort studies enrolled nonsmoking, healthy, and nonoccupationally exposed women and their newborns. Personal air monitoring of pregnant women was conducted over 48 hr. To control for maternal environmental tobacco smoke (ETS) exposure, we excluded those with umbilical cord plasma cotinine concentrations > 25 ng/mL. Mean cord plasma cotinine concentrations in both ethnic groups were 100-fold range) in lipid-adjusted levels for PBDEs in the general population. It is hypothesized that both exposure and pharmacokinetics may play a role in observed congener profiles. Significant gaps in our ability to interpret PBDE biomonitoring data to address public health and risk assessment questions include limited knowledge of environmental fate and transport of PBDE congeners, limited population-based data for adults, and lack of data for potentially vulnerable populations such as children. PMID- 17107867 TI - The applicability of biomonitoring data for perfluorooctanesulfonate to the environmental public health continuum. AB - Perfluorooctanesulfonate and its salts (PFOS) are derived from perfluorooctanesulfonyl fluoride, the basic chemical building block for many sulfonyl-based fluorochemicals used as surfactants and for their repellent properties. PFOS is highly persistent in the environment and has a long serum elimination half-life in both animals and humans. PFOS has been detected globally in the environment and in blood serum in various populations throughout the world, with the majority of human sampling done in the United States and Japan. The mechanisms and pathways leading to the presence of PFOS in human blood are not well characterized but likely involve both direct exposures to PFOS or chemicals and materials that can degrade to PFOS, either in the environment or from industrial and commercial uses. In 2000 the 3M Company, a major manufacturer, announced a phaseout of PFOS-related materials. Animal studies indicate that PFOS is well absorbed orally and distributes mainly in blood serum and the liver. Several repeat-dose toxicology studies in animals consistently demonstrated that the liver is the primary target organ. In addition there is a steep dose response for mortality in sexually mature rats and primates as well as in neonatal rats and mice exposed in utero. Several biomonitoring research needs that have been identified on PFOS include additional data from general populations pertaining to other matrices besides blood; matched serum and urine samples from humans and research animals; and comparison of whole blood, serum, and plasma concentrations from the same individuals. PMID- 17107868 TI - Integrating biomonitoring exposure data into the risk assessment process: phthalates [diethyl phthalate and di(2-ethylhexyl) phthalate] as a case study. AB - The probability of nonoccupational exposure to phthalates is high given their use in a vast range of consumables, including personal care products (e.g., perfumes, lotions, cosmetics), paints, industrial plastics, and certain medical devices and pharmaceuticals. Phthalates are of high interest because of their potential for human exposure and because animal toxicity studies suggest that some phthalates affect male reproductive development apparently via inhibition of androgen biosynthesis. In humans, phthalates are rapidly metabolized to their monoesters, which can be further transformed to oxidative products, conjugated, and eliminated. Phthalate metabolites have been used as biomarkers of exposure. Using urinary phthalate metabolite concentrations allows accurate assessments of human exposure because these concentrations represent an integrative measure of exposure to phthalates from multiple sources and routes. However, the health significance of this exposure is unknown. To link biomarker measurements to exposure, internal dose, or health outcome, additional information (e.g., toxicokinetics, inter- and intraindividual differences) is needed. We present a case study using diethyl phthalate and di(2-ethylhexyl) phthalate as examples to illustrate scientific approaches and their limitations, identify data gaps, and outline research needs for using biomonitoring data in the context of human health risk assessment, with an emphasis on exposure and dose. Although the vast and growing literature on phthalates research could not be covered comprehensively in this article, we made every attempt to include the most relevant publications as of the end of 2005. PMID- 17107869 TI - Biomarkers of exposure: a case study with inorganic arsenic. AB - The environmental contaminant inorganic arsenic (iAs) is a human toxicant and carcinogen. Most mammals metabolize iAs by reducing it to trivalency, followed by oxidative methylation to pentavalency. iAs and its methylated metabolites are primarily excreted in urine within 4-5 days by most species and have a relatively low rate of bioaccumulation. Intra- and interindividual differences in the methylation of iAs may affect the adverse health effects of arsenic. Both inorganic and organic trivalent arsenicals are more potent toxicants than pentavalent forms. Several mechanisms of action have been proposed for arsenic induced toxicity, but a scientific consensus has not been achieved. Biomarkers of exposure may be used to quantify exposure to iAs. The most common biomarker of exposure for iAs is the measurement of total urinary arsenic. However, consumption of seafood containing high concentrations of organic arsenic can confound estimation of iAs exposure. Because these organic species are thought to be relatively nontoxic, their presence in urine may not represent increased risk. Speciation of urinary arsenic into inorganic and organic forms, and even oxidation state, gives a more definitive indication of the exposure to iAs. Questions still remain, however, as to how reliably the measurement of urinary arsenic, either total or speciated, may predict arsenic concentrations at target tissues as well as how this measurement could be used to assess chronic exposures to iAs. PMID- 17107870 TI - Use of biomonitoring data to evaluate methyl eugenol exposure. AB - Methyl eugenol is a naturally occurring material found in a variety of food sources, including spices, oils, and nutritionally important foods such as bananas and oranges. Given its natural occurrence, a broad cross-section of the population is likely exposed. The availability of biomonitoring and toxicology data offers an opportunity to examine how biomonitoring data can be integrated into risk assessment. Methyl eugenol has been used as a biomarker of exposure. An analytical method to detect methyl eugenol in human blood samples is well characterized but not readily available. Human studies indicate that methyl eugenol is short-lived in the body, and despite the high potential for exposure through the diet and environment, human blood levels are relatively low. The toxicology studies in animals demonstrate that relatively high-bolus doses administered orally result in hepatic neoplasms. However, an understanding is lacking regarding how this effect relates to the exposures that result when food containing methyl eugenol is consumed. Overall, the level of methyl eugenol detected in biomonitoring studies indicates that human exposure is several orders of magnitude lower than the lowest dose used in the bioassay. Furthermore, there are no known health effects in humans that result from typical dietary exposure to methyl eugenol. PMID- 17107871 TI - 50 years Nobel Prize: Werner Forssmann and the issue of commemorative stamps. PMID- 17107872 TI - Werner Forssmann, Eberswalde, the 1956 Nobel Prize for medicine. AB - Since October 1949 Werner Forssmann was a regular guest of Prof. Dr. Hugo Wilhelm Knipping in the Medical University Clinic of Cologne. Established himself as urologist in Bad Kreuznach, Werner Forssmann had read about the American further development of heart catherization, which was invented by himself. Prof. Wilhelm Bolt, who was one of the medical station doctors of the Cologne Clinic, had already learned the technique of heart catherization in 1947. Thus, it was routinely performed in patients at the Cologne University Hospital. A close collaboration between Werner Forssmann and our research group (Hugo Wilhelm Knipping, Wilhelm Bolt, Helmut Valentin, Helmut Venrath, Hans Rink, Wildor Hollmann) was established. After the notification that Werner Forssmann had been awarded the Nobel Prize for Medicine in 1956, Hugo Wilhelm Knipping instructed me to help Werner Forssmann with the preparation of his lecture. Details of events in the year 1956 are illustrated. One of the important developments in which Werner Forssmann participated with the Medical University Clinic of Cologne was the selective pulmonary angiography. PMID- 17107873 TI - Recollection about Werner Forssmann as a surgeon. AB - The impact of Werner Forssmann on his clinical team during his time at the Protestant Hospital in Duesseldorf is described. Many residents came from foreign countries, especially from Argentina. The daily work of the clinics is depicted and the character of Werner Forssmann as a surgeon is understood from his education. PMID- 17107874 TI - When cardiology became a separate matter. AB - The cardiac catheter technique established by Werner Forssmann in 1929 not only introduced the era of modern cardiology, but also led to the identification of this specialist field and its practising doctors, the cardiologists. The intentionally scientifically oriented Deutsche Gesellschaft fur Kreislaufforschung, founded in 1927, acquired a clinical focus with the Commission for Clinical Cardiology thanks to the cardiac catheter cardiologists trained mainly in other countries. Today this society is the German Society for Cardiology, subtitled Cardiac and Cardiovascular Research, with a membership of 5,339 (July 31st 2004). Simultaneously, the impulse for the specialist fields to become independent within the framework of the new advanced educational regulations came into force in 1972 leading to the first branches of internal medicine, cardiology, gastroenterology and pneumology, followed by the establishment of independent departments, clinics and university chairs in cardiology. (At the end of 2003 a total of 3,059 internal specialists in cardiology were working in Germany and there were 393 independent facilities, including 33 university chairs in cardiology.) In addition, 10-14 cardiac centres combining both adult and pediatric cardiology together with cardiac surgery. When setting up intensive care wards, the main impulse also came from cardiology, achieving a totally new dimension when coronary angiography and bypass surgery made it possible to make an exact diagnosis as well as to provide an efficient therapy, right up to interventional methods with acute PTCA in recent cardiac infarction. The necessary step towards informing the population led to the establishment of the German Heart Foundation in 1979 with a membership of 46,795 (December 31st 2003) and a scientific board of 436 highly qualified cardiologists. The development presented here would not have been possible without the cardiac catheter. PMID- 17107875 TI - S100A1 gene transfer in myocardium. AB - S100A1, a Ca superset2+-binding protein of the EF-hand type, is preferentially expressed in myocardial tissue and has been shown to enhance cardiac contractile performance by regulating both sarcoplasmic reticulum (SR) Ca superset2+-handling and myofibrillar Ca superset2+-responsiveness. In cardiac disease, the expression of S100A1 is dynamically altered as it is significantly down-regulated in end stage human heart failure (HF), and it is up-regulated in compensated hypertrophy. Therefore, the delivery of a transgene encoding for S100A1 to the myocardium might be an attractive strategy for improving cardiac function in HF by replacing lost endogenous S100A1. In this study we sought to test whether exogenous S100A1 gene delivery to alter global cardiac function is feasible in the normal rabbit heart. An adenoviral S100A1 transgene (AdvS100A1) also containing the green fluorescent protein (GFP) was delivered using an intracoronary injection method with a dose of 5 x 10 superset11 total virus particles (tvp) (n = 8). Rabbits treated with either a GFP-only adenovirus (AdvGFP) or saline were used as control groups (n = 11 each). Seven days after global myocardial in vivo gene delivery hemodynamic parameters were assessed. S100A1 overexpression as a result of the intracoronary delivery of AdvS100A1 significantly increased left ventricular (LV) +dP/dt subsetmax, -dP/dt subsetmin and systolic ejection pressure (SEP) compared to both control groups after administration of isoproterenol (0.1, 0.5 and 1.0 microg/kgBW/min), while contractile parameters remained unchanged under basal conditions. These results demonstrate that global myocardial in vivo gene delivery is possible and that myocardial S100A1 overexpression can increase cardiac performance. Therefore, substitution of down-regulated S100A1 protein expression levels may represent a potential therapeutic strategy for improving the cardiac performance of the failing heart. PMID- 17107876 TI - Percutaneous septal ablation in hypertrophic obstructive cardiomyopathy. AB - This article reviews etiology, pathogenesis, natural history, and current treatment options in hypertrophic obstructive cardiomyopathy with a special focus on percutaneous septal ablation which has evolved as an alternative to myectomy in patients with symptoms refractory to medical treatment. Literature data and the own series of 337 interventions planned and 312 procedures completed (in hospital mortality and pacemaker implantation rates: 1.2% (4/312) and 7% (22/312)) are discussed. Overall satisfactory clinical results were seen in about 90% of the patients treated with septal ablation. PMID- 17107877 TI - AICD treatment in 2004--state of the art. AB - Primary and secondary prevention of sudden cardiac death is not sufficiently assured by medication. The (automatic) implantable cardioverter/defibrillator ((A)ICD) is able to terminate life-threatening arrhythmias (ventricular fibrillation/flutter, ventricular tachycardia) reliably. The identification and care of risk patients is of crucial importance. Initially, only survived resuscitation for ventricular fibrillation or ventricular tachycardia was regarded as a confirmed indication. Several studies (CABG patch, MADIT, MADIT II, MUSTT, DINAMIT, CAT AMIOVIRT, DEFINITE, COMPANION, SCD-HeFT) have examined the prophylactic indication for ICD therapy in risk groups. Patients with chronic state after myocardial infarction with markedly impaired left ventricular function and/or spontaneous, non-sustained ventricular tachycardia have been documented to benefit. Patients with moderately severe or severe heart failure also profit from ICD implantation, where appropriate in combination with cardiac resynchronization therapy in conduction disorders. There is divergent data on dilated cardiomyopathy. ICD is not indicated in patients with acute infarctions or undergoing elective bypass surgery. PMID- 17107878 TI - Stem cell therapy: a new perspective in the treatment of patients with acute myocardial infarction. AB - Experimental studies suggest that cardiac transfer of stem and progenitor cells can have a favorable impact on tissue perfusion and contractile performance after acute myocardial infarction (AMI). While the mechanistic background of stem cell therapy is still intensely debated, the concept of cell therapy has already been introduced into the clinical setting, where small, mostly uncontrolled trials indicate that stem cell therapy may be feasible in patients. The overall clinical experience also suggests that stem cell therapy can be safely performed, if the right cell type is used in the right clinical setting. Preliminary efficacy data indicate that stem cells have the potential to enhance myocardial perfusion and/or contractile performance in patients with AMI. The field now is rapidly moving towards intermediate-size, double-blinded trials to gather more safety and efficacy data. Ultimately, large outcome trials will have to be conducted. At the same time, continued basic research to elucidate the underlying mechanism of stem cell therapy is needed. PMID- 17107879 TI - ANP and urodilatin: who is who in the kidney. AB - Mounting evidence suggests that urodilatin, not atrial natriuretic peptide (ANP) is the responsible peptide in regulation of renal Na superset+- and water homeostasis. Following the discovery of ANP this peptide was thought to be responsible for the induction of natriuresis and diuresis in the mammalian kidney. However, the isolation of urodilatin from human urine and substantial work contributed to a better understanding of the renal physiology of these two natriuretic peptides. Indeed, subsequent elucidation supported that urodilatin rather than ANP seems to be the natriuretic peptide responsible for the regulation of Na superset+- and water homeostasis in the kidney. Urodilatin - synthesized and secreted from the distal tubules of the kidney - may act as a paracrine mediator when secreted into the lumen. In contrast, while the role of ANP as regulator of the cardiovascular system is established, its physiological regulatory role on transport processes in the nephron is questionable. This review attempts to analyze the roles of both ANP and urodilatin and to discuss new potential candidates which may also play a role in electrolyte and water handling in the kidney. PMID- 17107880 TI - Structure, function and amyloidogenic propensity of apolipoprotein A-I. AB - Apolipoprotein A-I, the major structural apolipoprotein of high-density lipoproteins, efficiently protects humans from cholesterol accumulation in tissues; however, it can cause systemic amyloidosis in the presence of peculiar amino acid replacements. The wild-type molecule also has an intrinsic tendency to generate amyloid fibrils that localise within the atherosclerotic plaques. The structure, folding and metabolism of normal apolipoprotein A-I are extremely complex and as yet not completely clarified, but their understanding appears essential for the elucidation of the amyloid transition. We reviewed present knowledge on the structure, function and amyloidogenic propensity of apolipoprotein A-I with the aim of highlighting the possible molecular mechanisms that might contribute to the pathogenesis of this disease. Important clues on apolipoprotein A-I amyloidogenesis may be obtained from classical comparative studies of the properties of the wild-type versus the amyloidogenic counterpart. Additionally, in the case of apoA-I, further insights on the molecular mechanisms underlying its amyloidogenic propensity may derive from comparative studies between amyloidogenic variants and other mutations associated with hypoalphalipoproteinemia without amyloidosis. PMID- 17107881 TI - Controlling amyloid growth in multiple dimensions. AB - The great progress made in defining the structure of protein and peptide amyloid assemblies, particularly the arrangement of peptides in beta-sheets, is counterbalanced by the still poor understanding of the higher organization of beta-sheets within the fibril and overall fibril/fibril associations. The assembly pathway and basis of amyloid toxicity may well depend on these higher order structural features. For example, significant evidence points to association between sheets as the rate limiting step in fibril assembly, and a critical metal binding site has now been identified that involves residues from different individual sheets. Here we review experiments that are identifying some of the issues associated with sheet-sheet association by investigating simple model peptides derived from the central core of the Abeta peptide implicated in Alzheimer's disease. These peptides transit between fibril/ribbon/nanotube morphologies in response to assembly conditions, laying the foundation for understanding the folding landscape for these higher order assemblies, revealing potential targets for therapeutic intervention, and opening strategies for the design of highly ordered peptide self-assembled microscale morphologies. PMID- 17107882 TI - The early stages of amyloid formation: biophysical and structural characterization of human calcitonin pre-fibrillar assemblies. AB - Amyloid fibril formation is a nucleation dependent process characterized by a lag phase prior to the appearance of detectable amyloid fibrils. While the three dimensional structure of amyloid fibrils at atomic resolution is just beginning to be elucidated, the early process of monomers assembly into oligomers is less understood. Understanding the dynamic processes that lead to the formation of these intermediates is highly important as these assemblies might be the most pathological ones. Here, we investigated the biophysical and structural features characterizing the early stage assemblies formed by the human hormone calcitonin. We calculated the initial nucleus size by experimentally determining the dependence between the lag-time length and the hCT concentrations. We used size exclusion chromatography and dynamic light scattering in order to characterize the dynamic growth process of preliminary intermediates transformed into larger structures. The early structures were visualized using high-resolution transmission electron microscopy. Annular pore-like structures were observed along with protofibrilar structures. This observed morphology is similar to structures revealed during the fibrillization processes of beta-amyloid, alpha synuclein, and islet amyloid polypeptide, suggesting that these intermediates represent a generic early structure conformation. The results introduced here imply that a variety of intermediate assemblies are formed during the early stages of amyloid fibril formation. The characterizing of their structural features and assembly kinetics will contribute to the rational design of inhibitors directed towards early structure assemblies. PMID- 17107883 TI - FALS mutations in Cu, Zn superoxide dismutase destabilize the dimer and increase dimer dissociation propensity: a large-scale thermodynamic analysis. AB - Mutations in the dimeric enzyme Cu, Zn superoxide dismutase (SOD1) leading to its aggregation are implicated in the toxicity in familial amyotrophic lateral sclerosis (FALS). We and others have previously shown that aggregation occurs by a pathway involving dimer dissociation, metal-loss from monomers and multimeric assembly of apo-SOD1 monomers. We postulate that FALS mutations cause enhanced aggregation by affecting one or more steps in the pathway, and computationally test this postulate for 75 known mis-sense FALS mutants of SOD1. Based on an extensive thermodynamic analysis of the stability of apo-dimer and apo-monomer forms of these mutants, we classify the mutations into the following groups: 70 out of 75 mutations in SOD1 lead to (i) decreased dimer stability, and/or (ii) increased dimer dissociation, compared to wild type, and four mutations lead to (iii) decreased monomer stability compared to wild type. Our results suggest that enhanced aggregation of SOD1 in FALS occurs due to an increased population of mutant SOD1 apo-monomers compared to wild type. The dissociation of multimeric proteins induced by diverse mutations may be a common theme in several human diseases. PMID- 17107884 TI - Orally administered diflunisal stabilizes transthyretin against dissociation required for amyloidogenesis. AB - OBJECTIVE: Rate-limiting transthyretin (TTR) tetramer dissociation and monomer misfolding enable misassembly into numerous aggregate morphologies including amyloid, a process genetically linked to and thought to cause amyloid pathology. T119M TTR trans-suppressor subunit inclusion into tetramers otherwise composed of disease-associated subunits ameliorates human amyloidosis by increasing the tetramer dissociation barrier. Diflunisal binding to the 99% unoccupied L thyroxine binding sites in TTR also increases the tetramer dissociation barrier; hence, we investigated the feasibility of using diflunisal for the treatment of human TTR amyloidosis using healthy volunteers. METHODS: Diflunisal (125, 250 or 500 mg bid) was orally administered to groups of 10 subjects for 7 days to evaluate serum diflunisal concentration, diflunisal binding stoichiometry to TTR, and the extent of diflunisal imposed TTR kinetic stabilization against urea- and acid-mediated TTR denaturation in human serum. The rates of urea-mediated tetramer dissociation and acid-mediated aggregation as a function of diflunisal concentration were also evaluated in vitro, utilizing physiologically relevant concentrations identified by the above experiments. RESULTS: In the 250 mg bid group, 12 h after the 13th oral dose, the diflunisal serum concentration of 146 +/- 39 microM was sufficient to afford a TTR binding stoichiometry exceeding 0.95 +/- 0.13 ( approximately 1.75 corrected). Diflunisal binding to TTR at this dose slowed urea-mediated dissociation and acid-mediated TTR aggregation at least, threefold (p < 0.05) in serum and in vitro, consistent with kinetic stabilization of TTR. CONCLUSION: Diflunisal-mediated kinetic stabilization of TTR should ameliorate TTR amyloidoses, provided that the nonsteroidal anti-inflammatory drug liabilities can be managed clinically. PMID- 17107885 TI - An in vitro model of early islet amyloid polypeptide (IAPP) fibrillogenesis using human IAPP-transgenic mouse islets. AB - The mechanisms underlying insufficient insulin secretion and loss of beta-cell mass in feline and human type 2 diabetes mellitus are incompletely understood. However, islet amyloid polypeptide (IAPP)-derived islet amyloidosis (IA) has been linked to increased rates of beta-cell apoptosis and, therefore, our goal was to develop an in vitro model of IAPP fibrillogenesis using isolated pancreatic islets from mice transgenic for human IAPP (hIAPP Tg mice). Islets from hIAPP Tg mice, from mice transgenic for non-amyloidogenic murine IAPP (mIAPP Tg mice), and from the FVB background strain were exposed to normal (5.5 mM) or high (28 mM) glucose conditions in cell culture for 8 days. On days 0 and 8, islets were collected for electron microscopy (EM). EM showed no abnormalities in the mIAPP Tg or FVB islets at either time point. On day 8, hIAPP Tg islets cultured at high glucose concentration formed extracellular IAPP-derived flocculent deposits. No significant differences in rates of apoptosis were found between groups. Our findings, therefore, show that in vitro culture of hIAPP Tg mouse islets under high glucose conditions produces a readily available and rapidly inducible model of IAPP-derived fibrillogenesis and enables the study of early phases of the molecular pathogenesis of IA. PMID- 17107886 TI - The amino acid sequence of an AL-protein, AL-KH, isolated from the heart of a patient with Waldenstroms macroglobulinemia and amyloidosis. AB - An AL-protein was isolated from the myocardium of a patient (KH) with Waldenstrom's macroglobulinemia. SDS-PAGE of the fraction revealed three major bands ranging from 14 to 30 kDa. Two of the major bands showed a positive PAS test for carbohydrate. The amino acid sequence was established for positions 1-92 and for positions 104-205, except for 150-187. According to the data, the AL protein KH is derived from the kappa 1c germline gene. PMID- 17107890 TI - Prognostic factors in chronic lymphocytic leukemia. PMID- 17107891 TI - Multi-drug resistance in B-cell chronic lymphocytic leukemia (B-CLL): A feature of B-CLL sub-sets with poor prognosis genetic alterations? PMID- 17107892 TI - Second neoplasms in chronic lymphocytic leukemia: Are case-group studies useful? PMID- 17107893 TI - The role of anthracyclines in follicular lymphomas. PMID- 17107894 TI - HFE H63D variant and leukemia susceptibility. PMID- 17107895 TI - Is there still place for VAD as primary treatment for patients with multiple myeloma who are candidates for high-dose therapy? PMID- 17107896 TI - Myeloma, thalidomide and thrombosis. PMID- 17107897 TI - The troublesome toxicity of peripheral neuropathy with thalidomide. PMID- 17107898 TI - A role for aberrant Notch signaling in acute myeloid leukemia? PMID- 17107899 TI - Optimizing treatment for elderly patients with acute promyelocytic leukemia: is it time to replace chemotherapy with all-trans retinoic acid and arsenic trioxide? AB - This review focuses on the treatment of acute promyelocytic leukemia (APL) in elderly patients and offers recommendations for improving outcomes. Nineteen percent of patients with APL are > or =60 years. Rates of response and survival are lower in elderly compared with younger patients, owing to a higher incidence of early deaths or deaths in remission. However, relapse-free survival rates are similar in both groups. Ongoing trials assess the role of reduced-intensity regimens. All-trans retinoic acid (ATRA) and concurrent arsenic trioxide is associated with high rates of response and molecular remission and low rates of induction deaths. We propose this combination as the treatment of choice in patients with APL, including the elderly. Patients with elevated leukocyte counts may also benefit from gemtuzumab ozogamicin therapy, with or without leukapheresis. Monitoring major organ function and toxicity is essential. Patients should be assessed for minimal residual disease using polymerase chain reaction testing for promyelocytic leukemia-retinoic acid receptor alpha. If molecular relapse is evident, treatment with ATRA and idarubicin, with or without gemtuzumab ozogamicin, is recommended. PMID- 17107900 TI - Ten years and counting: so what do we know about t(4;14)(p16;q32) multiple myeloma. AB - Multiple myeloma is a genetically heterogenous disease with a wide variety of characterized genetic aberrations. Until recently, the impact of these aberrations on patient outcome was not known. However, in the last 5-10 years, several genetic markers have been linked to patient outcome. One of the strongest predictors of outcome identified to date is t(4;14)(p16;q32). Although this translocation is tightly linked to chromosome 13 deletions, another poor prognosis marker, it is becoming apparent that the translocation and not the deletion of 13 is the important factor. Unfortunately, despite the known association with outcome, an understanding of the mechanism(s) whereby the translocation contributes to developing and maintaining this aggressive form of myeloma remains elusive. PMID- 17107901 TI - 2-CdA in the treatment of hairy cell leukemia: a review of long-term follow-up. AB - Hairy cell leukemia is a rare, indolent, chronic lymphoproliferative disorder characterized by the proliferation of a malignant B-cell clone with irregular cytoplasmic projections. Treatment with purine analogs such as 2 chlorodeoxyadenosine (2-CdA) is associated with excellent remission rates and long-term survival. Although the majority of patients achieve a complete remission (CR), most have minimal residual disease detected by sensitive methods. Despite the long term disease-free survival, approximately 36% of patients relapse and many require further therapy. Repeat administration of 2-CdA is very effective in achieving a second CR. Recently, new agents such rituximab and BL22 were shown to be effective in the treatment of relapsed and refractory disease. PMID- 17107902 TI - MDR-1, but not MDR-3 gene expression, is associated with unmutated IgVH genes and poor prognosis chromosomal aberrations in chronic lymphocytic leukemia. AB - Two P-glycoprotein (P-gp) genes, MDR-1 (ABCB1) and MDR-3 (ABCB4), have been identified in humans. This study was designed to investigate whether associations exist between expression of MDR-1 and MDR-3 P-gp and other markers of poor prognosis and/or prior exposure to therapeutic agents in chronic lymphocytic leukemia (CLL). IgVH mutational status, gene usage, CD38 positivity, FISH analysis and clinical information were available on all patients. Twenty-one of 101 patients tested showed MDR-3 P-gp positivity. Associations with markers of poor prognosis or prior chemotherapy did not reach statistical significance, but MDR-3 P-gp positive patients had significantly shorter survivals than MDR-3 P-gp negative patients. MDR-1 P-gp expression (18/25) showed a strong association with unmutated IgVH genes and adverse prognosis cytogenetics (p = 0.015, p = 0.014, respectively), but was independent of prior exposure to chemotherapeutic agents. These results suggest a role for MDR-1 and MDR-3 in chemoresistant disease. This study highlights the value of determining MDR phenotype in CLL patients prior to treatment, to allow the design of novel drug regimens containing agents that reverse MDR function. PMID- 17107903 TI - Incidence of second neoplasia in patients with B-cell chronic lymphocytic leukemia treated with chlorambucil maintenance chemotherapy. AB - The aim of this retrospective study was to examine the impact of prolonged chlorambucil (CLB) therapy on the development of second neoplasia (SN) in 389 patients with B-CLL, comparing untreated cases with those receiving CLB as induction plus maintenance therapy. Fifty-nine SN cases were observed (15.1%) at a median follow-up of 79 months. SN occurrence was significantly related to Binet stage. No difference was detected between untreated and CLB treated cases neither in terms of SN incidence (12.2% vs 18.1%) nor in the median follow-up (81 vs 79.1 months). Moreover, SN free survival was not different between these two groups. Four out of 13 CLB treated patients (30.8%) developed s-MDS after a subsequent treatment with fludarabine plus cyclophosphamide (F + C). In conclusion, SN development is dependent on the length of follow-up rather than on therapy duration. F + C should be administered with caution after prolonged CLB therapy. PMID- 17107904 TI - The addition of oral idarubicin to a chlorambucil/dexamethasone combination has a significant impact on time to treatment failure but none on overall survival in patients with low grade non-Hodgkin's lymphoma: Results of the Scotland and Newcastle Lymphoma Group randomized NHL VIII trial. AB - Two hundred untreated patients with low grade NHL (KIEL), including 155 follicular NHL, were randomized to six courses of treatment with chlorambucil 20 mg m-2 for 3 days and dexamethasone 4 mg bd for 5 days (CD) vs the same regimen plus oral idarubicin 10 mg m-2 for 3 days (CID). Responding patients could be randomized to no further treatment or maintenance treatment for up to 36 months with alpha interferon. Complete remissions/CRu were more frequent in the CID arm (35% vs 24%) but the overall response rate was similar; 87/91 (96%) vs 86/92 (93%). Overall survival (OS) did not differ between the two arms. Time to treatment failure (TTTF) was prolonged in the CID arm, p = 0.03; median time 28 vs 19 months. TTTF for the B-cell follicular group alone was for CID (77 patients) 33 months vs 18 months for CD (78 patients). Interferon conferred no apparent benefit. The Follicular Lymphoma International Prognostic Index (FLIPI) is confirmed as a good predictor of risk groups including a group of 23% with shorter survival. The addition of the oral anthracycline, idarubicin, led to a significant improvement in TTTF with low toxicity. The use of radiotherapy in this sub-group may have contributed to this result. CID is a potential for combination with antibody therapy particularly in older patient groups. PMID- 17107905 TI - HFE gene mutations in patients with acute leukemia. AB - An increased incidence of HFE gene mutations has been described in hematologic malignancies. In the present study, we investigated the allelic frequency of HFE gene mutations in 154 adult patients with acute leukemia (AL) [107 acute myeloid leukemia (AML), 20 acute promyelocytic leukemia (APL) and 27 acute lymphoblastic leukemia (ALL)]. The allelic frequency of the H63D mutation was 29% in AL patients and 25% in the healthy controls [P = 0.41; odds ratio (OR) = 1.20; 95% confidence interval (CI) = 0.77 - 1.93]. No difference was found between controls and AML or APL patients, whereas the H63D mutation was significantly more frequent in ALL than controls (44% vs. 25%, P = 0.04; OR = 2.37; 95% CI = 1.05 - 5.36). The overall comparison of the mutation among the three subtypes of AL demonstrated a higher allelic frequency in ALL (P = 0.02). In conclusion, our data demonstrate a correlation between the presence of the H63D mutation and the occurrence of ALL in adult patients. PMID- 17107906 TI - The combination of cyclophosphomide, thalidomide and dexamethasone is an effective alternative to cyclophosphamide - vincristine - doxorubicin - methylprednisolone as induction chemotherapy prior to autologous transplantation for multiple myeloma: a case-matched analysis. AB - A retrospective case-matched study was conducted to compare the oral regimen CTD (cyclophosphamide - thalidomide - dexamethasone) and infusional CVAMP (cyclophosphamide - vincristine - doxorubicin - methylprednisolone) as induction therapy followed by autologous peripheral blood stem-cell transplantation (PBSCT) for newly diagnosed multiple myeloma patients. The response rate after three cycles of treatment was statistically higher with CTD (n = 27) compared to CVAMP (n = 27) (89% vs. 56%, P = 0.016). Toxicity studies showed more neutropenia (grade 3/4) (4% vs. 60%, P = 0.0002) with CVAMP and more thrombotic episodes with CTD (11% vs. 4%). CTD may emerge as the superior induction regimen prior to PBSCT, in terms of high efficacy and better tolerability. PMID- 17107907 TI - Prospective evaluation of low-dose warfarin for prevention of thalidomide associated venous thromboembolism. AB - Venous thromboemobolism (VTE) is an important complication of thalidomide therapy especially when it is combined with steroids or chemotherapy. Currently there is no consensus on the most appropriate prophylactic approach. We prospectively investigated the use of low-dose warfarin sodium in prevention of thalidomide associated VTE in patients receiving thalidomide-based combination therapies. Patients with multiple myeloma or chronic lymphocytic leukemia who were treated on thalidomide based-combination therapies were treated on low-dose warfarin (1 or 2 mg) continuously through the duration of their therapy. Among the 68 patients enrolled, four developed an episode of VTE, an overall incidence of 5.9% (odds = 0.063). Median duration of thalidomide therapy was 4 months. Low-does warfarin decreases the incidence of VTE compared to historical control and is an effective mechanism of prevention of VTE in thalidomide-based chemotherapy regimens. PMID- 17107908 TI - Pegfilgrastim to support CHOP-14 in elderly patients with non-Hodgkin's lymphoma. AB - This study investigated whether pegfilgrastim support would enable on-schedule delivery of dose-dense cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP-14) to elderly patients with non-Hodgkin's lymphoma (NHL). Thirty patients 60 years of age and older with aggressive NHL were evaluated after receiving up to six cycles of CHOP-14 supported with pegfilgrastim. The median age was 68 years (range 61 - 74). Forty-seven per cent of patients received full dose chemotherapy on schedule for all cycles (range 65 - 93). Chemotherapy was delayed in 10 patients and dose reduced in 15 patients. Hematological toxicity was the most common reason for delays and dose reduction. Six of nine patients (67%) achieved a peripheral blood CD34+ count of at least 20 cellsx106 L-1 on day 12 of cycle one. The delivery on schedule of dose-dense CHOP-14 to elderly patients with previously untreated aggressive NHL is safe and efficacious with once per cycle pegfilgrastim support. PMID- 17107909 TI - Clonal lymphocytic populations in Philadelphia-negative chronic myeloproliferative disorders: is the T-cell clonality of 'undetermined' significance (TCUS) linked to a worse clinical outcome? AB - This study examined the clonality of B- and T-cells by PCR in 83 patients with Philadelphia-negative myeloproliferative disorders (Ph-MPD), to investigate its clinical and morphological correlates. Clonal lymphocytic populations were found in 23% of patients (T: n = 20, B: n = 3), with no frequency differences between ET, CIMF and PV. At the presentation, patients with clonal bands were older (58.1+/-13.8 vs 47.5+/-14.6, p = 0.0039), but did not differ in other clinical parameters. After the median follow-up of 21 months they were less likely to be asymptomatic (11.8% vs 41.1%, p = 0.029). The T-cell clonality was the strongest predictor of the symptomatic last follow-up by discriminant function analysis, surpassing the patient's age. This surprising negative prognostic impact of lymphocyte clonality in Ph-MPD may result from this phenomenon to be a better measure of the 'hematopoietic biologic age' than the metrical age itself. PMID- 17107910 TI - Chromosomal imbalances in pediatric Burkitt-like lymphoma and review of the literature in relation to other germinal center derived B-cell tumors. AB - This study reports on the cytogenetic features of a novel case of pediatric Burkitt-like lymphoma (BLL), that adds to the three published. Four groups of cytogenetic abnormalities were detected in the present case: (1) imbalances shared by most germinal center (GC) derived B-cell tumors including BLL (+1q, 6q, -8p, +8q24 and +11); (2) imbalances already reported in adult but not in pediatric BLL cases and shared with most GC B-cell tumors (+7, -9p, -9q, +12q, 13q, +17, +19, -3 and -4); (3) imbalances already reported in pediatric but not in adult BLL cases and shared with some GC B-cell tumors (-2q); and (4) imbalances never described before in pediatric or adult BLL, but present in different GC B-cell tumors (-6p, -1p and -18q). In view of the paucity of pediatric BLL cases published, this report adds novel, relevant information on the molecular cytogenetic features of this rare tumor. PMID- 17107911 TI - Incidence rates of the major leukemia subtypes among US Hispanics, Blacks, and non-Hispanic Whites. AB - While leukemia rates are thought to be lower in South and Central America, no study has systematically investigated incidence rates of the leukemia subtypes among Hispanics in the U.S. This was a retrospective cohort study, using data from the Surveillance, Epidemiology, and End Results (SEER) Program of the National Cancer Institute, 1992 - 2001, to compare leukemia incidence rates as a function of race and ethnicity. It was found that in adults, Hispanics had lower incidence rates for each of the major types of leukemia as compared to non Hispanic Whites: For AML, elderly Whites had an incidence rate ratio (IRR) of 1.61 in comparison to Hispanics (p < 0.001) and 1.27 in comparison to Blacks (p < 0.001); for CML, the IRR among the elderly was 1.42 that of Hispanics (p < 0.001) and 1.22 that of Blacks (p = 0.003); and for CLL, the IRR was 2.31 times that of Hispanics (p < 0.001) and 1.48 times that of Blacks (p < 0.001). In ALL, however, Hispanics aged 0 - 19 had a significantly higher incidence rate than Whites and Blacks, with an IRR of 1.32 compared to Whites (p < 0.001), and 2.62 compared to Blacks (p < 0.001). In AML, CML, and CLL, among people age 65 or older, white non Hispanics have higher incidence rates than Blacks, and Blacks have higher incidence rates than Hispanics. Childhood ALL incidence rates are highest among Hispanics, and lowest among Blacks. PMID- 17107912 TI - CD38 expression level and pattern of expression remains a reliable and robust marker of progressive disease in chronic lymphocytic leukemia. AB - Chronic lymphocytic leukemia (CLL) follows a variable clinical course which is difficult to predict at diagnosis. We assessed somatic mutation (SHM) status, CD38 and ZAP-70 expression in 87 patients (49 male, 38 female) with stage A CLL and known cytogenetic profile to compare their role in predicting disease progression, which was assessed by the treatment free interval (TFI) from diagnosis. Sixty (69%) patients were SHM+, 24 (28%) were CD38+ and ten (12%) were ZAP-70+. The median TFI for: (i) SHM + versus SHM- patients was 124 versus 26 months; hazard ratio (HR) = 3.6 [95% confidence interval (CI) = 1.8 - 7.3; P = 0.001]: (ii) CD38- versus CD38+ patients was 120 versus 34 months; HR = 2.4 (95% CI = 1.4 - 5.3; P = 0.02); and (iii) ZAP70- versus ZAP70+ was 120 versus 16 months; HR = 3.4 (95% CI = 1.4 - 8.7; P = 0.01). SHM status and CD38 retained prognostic significance on multivariate analysis whereas ZAP-70 did not. We conclude that ZAP-70 analysis does not provide additional prognostic information in this group of patients. PMID- 17107913 TI - Anti-CD19 immunotoxin enhances the activity of chemotherapy in severe combined immunodeficient mice with human pre-B acute lymphoblastic leukemia. AB - The anti-CD19 immunotoxin (IT) (HD37-dgRTA) is effective in killing B-lineage leukemia cells and in curing severe combined immunodeficient mice with acute lymphoblastic leukemia. The present study aimed to identify effective combinations of HD37-dgRTA and chemotherapeutic agents. The in-vitro cytotoxicity assays demonstrate that the combination of HD37-dgRTA and either daunorubicin or vincristine is effective. The in-vivo experiments using HD37-dgRTA with vincristine prolonged the survival of mice compared to the chemotherapeutic agent or IT (90.7 vs. 147.1 days). Also, 80% of the mice treated with IT plus vincristine were long-term survivors. PMID- 17107914 TI - Characterization of antibody-containing vesicles shed from B-lymphoma cell lines: exposure of annexin V binding sites. AB - Antibodies (Abs) to CD20 or HLA-DR, after binding to the B-lymphoma cell line RL following an overnight incubation at 37 degrees C, accumulate in the form of shed vesicles, which develop in the center of the cell clusters that are spontaneously formed by this cell line. These vesicles coalesce into fairly stable large structures, which we refer to as conglomerates of shed vesicles (CSVs). In the present study, we have extended our previous investigations into the nature of this material. Electron microscopy revealed a conglomerate of heterogeneous vesicles, which looked like pinched-off cytoplasmic projections. CSVs developed similarly either with or without Ab, demonstrating that CSV production is a spontaneous process that incorporates bound Abs if they are present. Before delivery to CSVs, the Abs capped on the cell surface. CSVs had high expression of annexin V binding sites, which are phagocytic signals that are exposed on damaged cells. For CSVs that were cell bound, which are frequently observed, the annexin V binding sites were only in the CSVs, and not on the surface of the intact cell. Although all CSVs contained both Abs and annexin V binding sites, the precise distribution of these two ligands was generally different. Annexin V binding sites were present on caps as well as on CSVs, and appear as soon as caps are formed. In cells incubated with anti-HLA-DR, CD20 was delivered to the CSVs together with HLA-DR, suggesting an association between these two molecules. CSVs prepared with anti-HLA-DR, but not CSVs prepared with anti-CD20, contained considerable numbers of nuclear fragments, identified by propidium iodide staining. PMID- 17107915 TI - NOTCH1 mutations are rare in acute myeloid leukemia. AB - Mutations in the NOTCH1 gene were investigated in 12 primary acute myeloid leukemia (AML) cell samples and eight AML cell lines. Mutations in the genomic DNA were screened using a nested PCR-SSCP analysis and confirmed by direct sequencing. A missense mutation, Pro2439Leu (7316C/T), was found in the PEST domain in one primary AML case. This mutation was different from those previously reported for T-cell acute lymphoblastic leukemia, in which more than half the cases had the mutations. This mutation was not detected in his sample in complete remission, which indicated that the mutation was not a single nucleotide polymorphism. The sample with the mutation expressed the intracellular Notch1 fragment by immunoblotting and HES1 mRNA by reverse transcription-polymerase chain reaction. This is the first paper to present an AML case with NOTCH1 mutation. The precise role of the mutation is to be determined. PMID- 17107916 TI - Case 31: Eosinophilia and pruritus. AB - A 39-year-old man presented with a pruritic rash, abdominal pain, weight loss and eosinophilia. A subsequent emergency laparotomy disclosed the nature of his underlying illness and the cause of the eosinophilia. PMID- 17107917 TI - Lymphoplasmacytic lymphoma presenting as malignant ascites. PMID- 17107918 TI - The efficacy of low dose thalidomide in refractory/relapsed myeloma: a retrospective audit. PMID- 17107919 TI - Can survivin expression predict the response to bortezomib in cases with mantle cell lymphoma? PMID- 17107920 TI - Megakaryoblastic differentiation of myeloid sarcoma in a patient with essential thrombocythemia. PMID- 17107921 TI - Bilateral thalamic infarction after reinfusion of DMSO-preserved autologous stem cells. PMID- 17107922 TI - Characteristics and outcome of B-cell chronic lymphocytic leukemia in hepatitis C virus-positive patients. PMID- 17107923 TI - Hepatocellular carcinoma with indolent T-lymphoblastic proliferation. PMID- 17107924 TI - Imatinib mesylate-induced long-term remission in extra-medullary T-cell lymphoid blastic phase of chronic myelogenous leukemia. PMID- 17107925 TI - Liposomes and micellar dispersions for delivery of benzoheterocyclic derivatives of distamycin A. AB - In this article we describe the production and characterization of specialized delivery systems for some distamycin derivatives (DD), namely liposomes and micellar dispersions. All the formulations were designed to increase the solubility of DD in an aqueous environment and to reduce the possible toxicity problems related to the administration of these drugs. For instance, liposomes were prepared by reverse phase evaporation technique followed by extrusion through polycarbonate filters, then characterized in terms of dimensions, morphology, and encapsulation efficacy. The analysis of their in vitro antiproliferative activity on cultured human and mouse leukemic cells demonstrated that liposomes and micellar dispersions containing DD exert quite different effects. These effects were compared with those shown by the free drug depending on type of drug and also cell line used. PMID- 17107926 TI - Synthesis and characterization of pH-sensitive thiol-containing chitosan beads for controlled drug delivery applications. AB - The aim of this study was to develop chitosan-based materials in drug delivery systems possessing covalent attachment of thiol moieties. Thiol-containing chitosan (TCS), found to be soluble in water, was synthesized by graft copolymerization technique. The TCS beads were prepared by using tripolyphoshate, at pH 4.0. The morphology of TCS beads was examined by scanning electron microscopy. The in vitro drug release behavior was studied in phosphate buffer solution at various pH, using indomethacin as a model drug at two different concentrations (0.3 and 0.6% w/w). The release amounts of indomethacin from TCS beads were higher increasing pHs in the dissolution medium. The release rate of indomethacin at pH 7.4 was higher than the release rate at pH 1.4 due to ionization of thiol groups and high solubility of indomethacin in an alkaline medium. These results indicated that the TCS beads may become a delivery system for the controlled release of different drugs wherever pH sensitive mechanics might be useful. This is especially applicable in cases when it is important to minimize drug release in acidic sites, such as in the stomach. PMID- 17107927 TI - Chitosan phthalate microspheres for oral delivery of insulin: preparation, characterization, and in vitro evaluation. AB - Chitosan phthalate polymer was synthesized and its microspheres were prepared by emulsion phase separation technique. The characterization of microspheres was determined by means of FTIR spectroscopy, electron microscopy, particle size, and zeta potential. The insulin was loaded to the microspheres by passive absorption technique. The peptic and tryptic enzymes degradation of insulin in microspheres was investigated. The in vitro release behavior of the microspheres was investigated under different pH conditions (pH 2.0 and pH 7.4). The degree of phthalate substitution in the synthesized polymer was 20%. The prepared microspheres were spherical with an average diameter 46.34 micro m. The insulin loading capacity was 62%. Chitosan phthalate microspheres protect the insulin from gastric enzymes degradation that may enhance the oral stability of insulin. The encapsulated insulin was quickly released in a phosphate buffer saline (pH 7.4), whereas a small amount of insulin was released under acidic condition (0.1N HCl; pH 2.0) because under acidic conditions, carboxylic groups present in the system exist in nonionized form and are poorly hydrophilic. However, in alkaline conditions, it exists in ionized form and is considerably hydrophilic. The results suggest that chitosan phthalate microspheres may be used as a potential carrier for oral insulin delivery. PMID- 17107929 TI - Improving the dissolution rate of poorly water soluble drug by solid dispersion and solid solution: pros and cons. AB - The solid dispersions with poloxamer 188 (P188) and solid solutions with polyvinylpyrrolidone K30 (PVPK30) were evaluated and compared in an effort to improve aqueous solubility and bioavailability of a model hydrophobic drug. All preparations were characterized by differential scanning calorimetry, powder X ray diffraction, intrinsic dissolution rates, and contact angle measurements. Accelerated stability studies also were conducted to determine the effects of aging on the stability of various formulations. The selected solid dispersion and solid solution formulations were further evaluated in beagle dogs for in vivo testing. Solid dispersions were characterized to show that the drug retains its crystallinity and forms a two-phase system. Solid solutions were characterized to be an amorphous monophasic system with transition of crystalline drug to amorphous state. The evaluation of the intrinsic dissolution rates of various preparations indicated that the solid solutions have higher initial dissolution rates compared with solid dispersions. However, after storage at accelerated conditions, the dissolution rates of solid solutions were lower due to partial reversion to crystalline form. The drug in solid dispersion showed better bioavailability in comparison to solid solution. Therefore, considering physical stability and in vivo study results, the solid dispersion was the most suitable choice to improve dissolution rates and hence the bioavailability of the poorly water soluble drug. PMID- 17107928 TI - Targeting the cerebrovascular large neutral amino acid transporter (LAT1) isoform using a novel disulfide-based brain drug delivery system. AB - We describe a novel strategy to achieve high affinity recognition for the specific, cerebrovascular large neutral amino acid transporter (LAT1) isoform by covalent coupling of small molecules to the amino acid, L-cysteine (L-Cys). L-Cys (as the carrier) was covalently attached via a disulfide bond to either 6 mercaptopurine or 2-methyl-1-propanethiol (IBM) to form the brain-targeted drug delivery systems (BTDS). BTDS were designed for high affinity recognition by LAT1 at the cerebrovasculature. Using an in situ rat brain perfusion technique, competition between BTDS and the radiotracer [14C]L-Leu demonstrated significant inhibition of [14C]L-Leu brain uptake. BTDS possess affinity for cerebrovascular LAT1 in many distinct brain compartments, and the recognition of BTDS by LAT1 is influenced by hydrophobicity of the side-chain in BTDS. Thus, the BTDS strategy may be utilized for rapid shuttling of various neuropharmaceuticals into brain. PMID- 17107930 TI - In vitro evaluation of enrofloxacin-loaded MLV liposomes. AB - Fluoroquinolones are broad-spectrum antimicrobial agents that seem to reach their intracellular target site (DNA gyrase) in Escherichia coli by means of an uptake process through the outer and inner membranes. Delivery of quinolones with liposomes has many advantages than the free form of the drug. Liposomes may represent an excellent device for improving the selective transport of antibiotics in these respects. In this study, enrofloxacin-loaded multilamellar vesicles (MLVs) were prepared and the effects of formulation variables on the liposome characteristics were investigated. Liposomes were prepared by using the dry lipid film method. A number of variables, such as phospholipid (DL-alpha phosphatidylcholine dipalmitoyl), cholesterol, enrofloxacin (ENF), stearylamine, and dicetyl phosphate molar ratios and alpha -tocopherol amounts, were studied. The liposome size, encapsulation capacity, drug release, stability, and electrophoretic mobility of ENF-loaded liposomes were determined. Using this method, spherical MLVs with high drug content could be produced. Particle size of liposomes changed between 1.63 and 3.31 micro m and liposome size was affected by all formulation variables (p < 0.05) except molar ratio of ENF. MLVs can be used as a carrier system for the controlled release of ENF. The highest encapsulation of ENF amount can be obtained using positively charged SA in the formulation and changing the formulation parameters can vary drug release patterns. PMID- 17107934 TI - The rhythms of life: circadian output pathways in Neurospora. AB - Research in Neurospora crassa pioneered the isolation of clock-controlled genes (ccgs), and more than 180 ccgs have been identified that function in various aspects of the fungal life cycle. Many clock-controlled genes are associated with damage repair, stress responses, intermediary metabolism, protein synthesis, and development. The expression of most of these genes peaks just before dawn and appears to prepare the cells for the desiccation, mutagenesis, and stress caused by sunlight. Progress on characterization of the output signaling pathways from the circadian oscillator mechanism to the ccgs is discussed. The authors also review evidence suggesting that, similar to other clock model organisms, a connection exists between the redox state of the cell and the Neurospora clock. The authors speculate that the clock system may sense not only light but also the redox potential of the cell through one of the PAS domains of the core clock components WC-1 or WC-2. PMID- 17107933 TI - No promoter left behind: global circadian gene expression in cyanobacteria. AB - Prokaryotic cyanobacteria express robust circadian (daily) rhythms under the control of a clock system that appears to be similar to those of eukaryotes in many ways. On the other hand, the KaiABC-based core cyanobacterial clockwork is clearly different from the transcription-translation feedback loop model of eukaryotic clocks in that the cyanobacterial clock system regulates gene expression patterns globally, and specific clock gene promoters are not essential in mediating the circadian feedback loop. A novel model, the oscilloid model, proposes that the KaiABC oscillator ultimately mediates rhythmic changes in the status of the cyanobacterial chromosome, and these topological changes underlie the global rhythms of transcription. The authors suggest that this model represents one of several possible modes of regulating gene expression by circadian clocks, even those of eukaryotes. PMID- 17107935 TI - Mechanisms of clock output in the Drosophila circadian pacemaker system. AB - Molecular oscillations that underlie the circadian clock are coupled to different output signals by which daily rhythms in downstream events are evoked and/or synchronized. Here the authors review the literature that describes circadian output mechanisms in Drosophila. They begin at the most proximal level, within oscillator cells themselves, by surveying studies of rhythmic gene expression within Drosophila heads. Next the authors describe the several neuron groups that compose the circadian pacemaker network underlying rhythmic locomotor activity, and they detail current models of how that network is organized and coordinated. The authors outline the body of evidence that describes a role for the neuropeptide pigment dispersing factor (PDF) as a circadian transmitter in the fly brain. Finally, in the context of PDF, they consider studies that address mechanisms of signaling from the circadian pacemaker network to downstream neurons and nonneuronal cells that directly control rhythmic outputs. PMID- 17107936 TI - SCN outputs and the hypothalamic balance of life. AB - The circadian clock in the suprachiasmatic nucleus (SCN) is composed of thousands of oscillator neurons, each dependent on the cell-autonomous action of a defined set of circadian clock genes. Still, the major question remains how these individual oscillators are organized into a biological clock producing a coherent output able to time all the different daily changes in behavior and physiology. In the present review, the authors discuss the anatomical connections and neurotransmitters used by the SCN to control the daily rhythms in hormone release. The efferent SCN projections mainly target neurons in the medial hypothalamus surrounding the SCN. The activity of these preautonomic and neuroendocrine target neurons is controlled by differentially timed waves of, among others, vasopressin, GABA, and glutamate release from SCN terminals. Together, the data on the SCN control of neuroendocrine rhythms provide clear evidence not only that the SCN consists of phenotypically (i.e., according to neurotransmitter content) different subpopulations of neurons but also that subpopulations should be distinguished (within phenotypically similar groups of neurons) based on the acrophase of their (electrical) activity. Moreover, the specialization of the SCN may go as far as a single body structure, that is, the SCN seems to contain neurons that specifically target the liver, pineal, and adrenal. PMID- 17107937 TI - Encoding the ins and outs of circadian pacemaking. AB - The SCN of the mammalian hypothalamus comprises a self-sustained, biological clock that generates endogenous ca. 24-h (circadian) rhythms. Circadian rhythmicity in the SCN originates from the interaction of a defined set of "clock genes" that participate in transcription/translation feedback loops. In order for the SCN to serve as an internal clock that times an internal day corresponding to the external solar day, the intracellular molecular oscillations must be output as physiological signals and be reset by appropriate environmental inputs. Here, the authors consider the mechanisms by which the SCN circadian pacemaker encodes rhythmic output and light input. In particular, they focus on the ionic mechanisms by which SCN neurons encode clock gene output as circa-dian rhythms in spike frequency, as well as cellular and molecular mechanisms by which SCN neurons encode circadian light input through phase heterogeneity in the SCN network. The authors propose that there are 2 distinct classes of ionic mechanisms supporting spike frequency rhythms output--modulation of basal membrane potential and conductance versus modulation of spike production--whereas light input is transformed by cellular communication within the SCN network and encoded by the relative phase relationships among SCN neurons. PMID- 17107938 TI - Neurobiology of the sleep-wake cycle: sleep architecture, circadian regulation, and regulatory feedback. AB - This mini-review article presents the remarkable progress that has been made in the past decade in our understanding of the neural circuitry underlying the regulation of sleep-wake states and circadian control of behaviors. Following a brief introduction to sleep architecture and physiology, the authors describe the neural circuitry and neurotransmitters that regulate sleep and cortical arousal (i.e., wakefulness). They next examine how sleep and wakefulness are regulated by mutual inhibition between sleep-and arousal-promoting circuitry and how this interaction functions analogously to an electronic "flip-flop" switch that ensures behavioral state stability. The authors then discuss the role of circadian and homeostatic processes in the consolidation of sleep, including the physiologic basis of homeostatic sleep drive (i.e., wake-dependent increase in sleep propensity) and the role of the SCN in the circadian regulation of sleep wake cycles. Finally, they describe the hypothalamic circuitry for the integration of photic and nonphotic environmental time cues and how this integration allows organisms to sculpt patterns of rest-activity and sleep-wake cycles that are optimally adaptive. PMID- 17107939 TI - Properties, entrainment, and physiological functions of mammalian peripheral oscillators. AB - In mammals, the circadian timing system is composed of multiple oscillators that are organized in a hierarchical manner. The central pacemaker, located in the suprachiasmatic nucleus of the hypothalamus, is believed to orchestrate countless subsidiary clocks in the periphery. These peripheral oscillators are cell autonomous, self-sustained, resilient to cell division, and virtually insensitive to large fluctuations in general transcription rates. However, they are probably not coupled within an organ, and daily zeitgeber signals emanating from the SCN appear to be required to ensure phase coherence within and between tissues. Peripheral clocks are implicated in a variety of biochemical pathways, and recent results tightly link circadian rhythms to several aspects of metabolism. Thus, the expression of many key enzymes conducting rate-limiting steps in various metabolic pathways is regulated in a circadian fashion by core clock components or clock-controlled transcription factors. Genetic loss-of-function studies have now established a role for mammalian circadian clock components in energy homeostasis and xenobiotic detoxification, and the latter manifests itself in the daytime-dependent modulation of drug efficacy and toxicity. PMID- 17107940 TI - Systems biology of circadian rhythms: an outlook. AB - The circadian system in higher organisms temporally orchestrates rhythmic changes in a vast number of genes and gene products in different organs. Complex interactions between these components, both within and among cells, ultimately lead to rhythmic behavior and physiology. Identifying the plethora of circadian targets and mapping their interactions with one another is therefore essential to comprehend the molecular mechanisms of circadian regulation. The emergence of new technology for unbiased identification of biomolecules and for mapping interactions at the genome-wide scale is offering powerful tools to decipher the regulatory networks underpinning circadian rhythms. In this review, the authors discuss the potential application of these genome-wide approaches in the study of circadian rhythms. PMID- 17107941 TI - Proteomics in renal research. AB - Proteomic technologies are used with increasing frequency in the renal community. In this review, we highlight the use in renal research of a number of available techniques including two-dimensional gel electrophoresis, liquid chromatography/mass spectrometry, surface-enhanced laser desorption/ionization, capillary electrophoresis/mass spectrometry, and antibody and tissue arrays. These techniques have been used to identify proteins or changes in proteins specific to regions of the kidney or associated with renal diseases or toxicity. They have also been used to examine protein expression changes and posttranslational modifications of proteins during signaling. A number of studies have used proteomic methodologies to look for diagnostic biomarkers in body fluids. The rapid rate of development of the technologies along with the combination of classic physiological and biochemical techniques with proteomics will enable new discoveries. PMID- 17107942 TI - Extracellular signal-regulated kinase activation by parathyroid hormone in distal tubule cells. AB - The PTH receptor (PTH1R) activates multiple signaling pathways, including extracellular signal-regulated kinases 1 and 2 (ERK1/2). The role of epidermal growth factor receptor (EGFR) transactivation in ERK1/2 activation by PTH in distal kidney cells, a primary site of PTH action, was characterized. ERK1/2 phosphorylation was stimulated by PTH and blocked by the EGFR inhibitor, AG1478. Upon PTH stimulation, metalloprotease cleavage of membrane-bound heparin-binding fragment (HB-EGF) induced EGFR transactivation of ERK. Conditioned media from PTH treated distal kidney cells activated ERK in HEK-293 cells. AG1478 added to HEK 293 cells ablated transactivation by conditioned media. HB-EGF directly activated ERK1/2 in HEK-293 cells. Pretreatment of distal kidney cells with the metalloprotease inhibitor GM-6001 abolished transactivation of ERK1/2 by PTH. The role of the PTH1R COOH terminus in PTX-sensitive ERK1/2 activation was characterized in HEK-293 cells transfected with wild-type PTH1R, with a PTH1R mutated at its COOH terminus, or with PTH1R truncated at position 480. PTH stimulated ERK by wild-type, mutated and truncated PTH1Rs 21-, 27- and 57-fold, respectively. Thus, the PTH1R COOH terminus exerts an inhibitory effect on ERK activation. EBP50, a scaffolding protein that binds to the PDZ recognition domain of the PTH1R, impaired PTH but not isoproterenol or calcitonin-induced ERK activation. Pertussis toxin inhibited PTH-stimulated ERK1/2 by mutated and truncated PTH1Rs and abolished ERK1/2 activation by wild-type PTH1R. We conclude that ERK phosphorylation in distal kidney cells by PTH requires PTH1R activation of G(i), which leads to stimulation of metalloprotease-mediated cleavage of HB EGF and transactivation of the EGFR and is regulated by EBP50. PMID- 17107943 TI - Angiotensin-converting enzyme inhibition aggravates renal interstitial injury resulting from partial unilateral ureteral obstruction in the neonatal rat. AB - Congenital urinary tract obstruction is the most important cause of renal insufficiency in infants and children, and angiotensin-converting enzyme (ACE) inhibitors attenuate the progression of renal disease in adults. ACE inhibitors are increasingly utilized in children with progressive renal disease. Because angiotensin is necessary for normal renal development, we examined the effects of ACE inhibition both during and immediately following the period of postnatal nephrogenesis in the neonatal rat subjected to sham operation or partial unilateral ureteral obstruction (UUO) under general anesthesia within the first 48 h of life. Rats in group I received enalapril 30 mg/kg body wt (or vehicle) daily for the first 10 days, while in group II, the 10 days of treatment began 10 days after surgery. Kidneys were harvested at day 21 and analyzed for apoptosis (TUNEL), interstitial macrophages (ED-1 immunohistochemistry), myofibroblasts (alpha-smooth muscle actin), and collagen (Sirius red). Partial UUO delayed glomerular maturation and increased ipsilateral renal macrophage infiltration, alpha-smooth muscle actin and Sirius red staining. In group I, enalapril increased myofibroblast accumulation in sham-operated kidneys, but not in obstructed kidneys. In contrast, in group II, enalapril further increased macrophage, myofibroblast, and collagen accumulation following partial UUO. The relative abundance of components of the kallikrein-kinin system, measured by Western blot, was not altered by partial UUO in the 14- and 28-day-old rat. Thus, in contrast to its salutary effects at later ages, ACE inhibition can worsen injury to the partially obstructed kidney during renal maturation even after the completion of nephrogenesis. PMID- 17107944 TI - Origin of spontaneous activity in neonatal and adult rat bladders and its enhancement by stretch and muscarinic agonists. AB - This study examined the origin of spontaneous activity in neonatal and adult rat bladders and the effect of stretch and muscarinic agonists and antagonists on spontaneous activity. Rats were anesthetized and their bladders were excised, cannulated, and loaded with voltage- and Ca(2+)-sensitive dyes. Intracellular Ca(2+) and membrane potential transients were mapped using photodiode arrays in whole bladders, bladder sheets, or cross-section preparations at 37 degrees C. Intravesical pressure was recorded from whole bladders. In neonatal bladders and sheets, spontaneous Ca(2+) and electrical signals arose at a site near the dome and spread in a coordinated manner throughout the bladder with different dome-to neck conduction velocities (Ca(2+): 3.7 +/- 0.4 mm/s; membrane potential: 46.2 +/ 3.1 mm/s). In whole bladders, optical signals were associated with spontaneous contractions (10-20 cmH(2)O). By contrast, in adult bladders spontaneous Ca(2+) and electrical activity was uncoordinated, originating at multiple sites and was associated with smaller (2-5 cmH(2)O) contractions. Spontaneous contractions and optical signals were insensitive to tetrodotoxin (2 muM) but were blocked by nifedipine (10 muM). Stretch or low carbachol concentrations (50 nM) applied to neonatal whole bladders enhanced the amplitude (to 20-35 cmH(2)O) of spontaneous activity, which was blocked by atropine. Bladder cross sections revealed that Ca(2+) and membrane potential transients produced by stretch or carbachol began near the urothelial-suburothelial interface and then spread to the detrusor. In conclusion, spontaneous activity in neonatal bladders, unlike activity in adult bladders, is highly organized, originating in the urothelium-suburothelium near the dome. Activity is enhanced by stretch or carbachol and this enhancement is blocked by atropine. It is hypothesized that acetylcholine is released from the urothelium during bladder filling to enhance spontaneous activity. PMID- 17107945 TI - Insertion of anthrax protective antigen into liposomal membranes: effects of a receptor. AB - Protective antigen (PA), the receptor-binding component of anthrax toxin, heptamerizes and inserts into the endosomal membrane at acidic pH, forming a pore that mediates translocation of the enzymic components of the toxin to the cytosol. When the heptameric pre-insertion form of PA (the prepore) is acidified in solution, it rapidly loses the ability to insert into membranes. To maximize insertion into model membranes, we examined two ways to bind the protein to large unilamellar vesicles (LUV). One involved attaching a His tag to the von Willebrand factor A domain of one of the PA receptors, ANTXR2, and using this protein as a bridge to bind PA to LUV containing a nickel-chelating lipid. The other involved using a His tag fused to the C terminus of PA to bind the protein directly to LUV containing the same lipid. Both ways enhanced pore formation at pH 5.0 strongly and about equally, as measured by the release of K+. Controls showed that pore formation in this system faithfully reproduced that in vivo. We also showed that binding unmodified ANTXR2 von Willebrand factor A to the prepore in solution enhanced its pore forming activity by slowing its inactivation at acidic pH. These findings indicate that an important role of PA receptors is to promote partitioning of PA into the bilayer by maintaining the prepore close to the target membrane and presumably in the optimal orientation as it undergoes the acidic pH-dependent conformational transition to the pore. PMID- 17107946 TI - A cadmium-transporting P1B-type ATPase in yeast Saccharomyces cerevisiae. AB - Detoxification and homeostatic acquisition of metal ions are vital for all living organisms. We have identified PCA1 in yeast Saccharomyces cerevisiae as an overexpression suppressor of copper toxicity. PCA1 possesses signatures of a P1B type heavy metal-transporting ATPase that is widely distributed from bacteria to humans. Copper resistance conferred by PCA1 is not dependent on catalytic activity, but it appears that a cysteine-rich region located in the N terminus sequesters copper. Unexpectedly, when compared with two independent natural isolates and an industrial S. cerevisiae strain, the PCA1 allele of the common laboratory strains we have examined possesses a missense mutation in a predicted ATP-binding residue conserved in P1B-type ATPases. Consistent with a previous report that identifies an equivalent mutation in a copper-transporting P1B-type ATPase of a Wilson disease patient, the PCA1 allele found in laboratory yeast strains is nonfunctional. Overexpression or deletion of the functional allele in yeast demonstrates that PCA1 is a cadmium efflux pump. Cadmium as well as copper and silver, but not other metals examined, dramatically increase PCA1 protein expression through post-transcriptional regulation and promote subcellular localization to the plasma membrane. Our study has revealed a novel metal detoxification mechanism in yeast mediated by a P1B-type ATPase that is unique in structure, substrate specificity, and mode of regulation. PMID- 17107947 TI - The liver X receptor (LXR) and hepatic lipogenesis. The carbohydrate-response element-binding protein is a target gene of LXR. AB - The liver X receptors, LXRalpha (NR1H3) and LXRbeta (NR1H2), are ligand-activated transcription factors that belong to the nuclear hormone receptor superfamily. LXRs play a critical role in cholesterol homeostasis and bile acid metabolism. In addition, oral administration of LXR agonists to mice results in elevated hepatic fatty acid synthesis and steatosis and increased secretion of triglyceride-rich very low density lipoprotein resulting in hypertriglyceridemia. This increased hepatic lipogenesis has been largely attributed to the LXR-dependent up regulation of sterol regulatory element-binding protein 1c (SREBP-1c) expression. However, it has been reported that treating Srebp-1c null mice with the synthetic LXR agonist T0901317 still results in enhanced expression of many lipogenic genes, suggesting additional mechanisms by which LXR can enhance hepatic lipogenesis. In this report, we identify the carbohydrate response element binding protein (ChREBP) as an LXR target that independently enhances the up regulation of select lipogenic genes. The ChREBP promoter contains functional LXR binding sites that confer receptor-dependent binding and transactivation. We show that T0901317 treatment of mice is associated with up-regulation of the ChREBP target gene, liver-type pyruvate kinase. Therefore, activation of LXR not only increases ChREBP mRNA via enhanced transcription but also modulates ChREBP activity. This establishes LXR as a master lipogenic transcription factor, as it directly regulates both SREBP-1c and ChREBP to enhance hepatic fatty acid synthesis. PMID- 17107948 TI - Biochemical and structural characterization of the gem GTPase. AB - RGK proteins, encompassing Rad, Gem, Rem1, and Rem2, constitute an intriguing branch of the Ras superfamily; their expression is regulated at the transcription level, they exhibit atypical nucleotide binding motifs, and they carry both large N- and C-terminal extensions. Biochemical and structural studies are required to better understand how such proteins function. Here, we report the first structure for a RGK protein: the crystal structure of a truncated form of the human Gem protein (G domain plus the first part of the C-terminal extension) in complex with Mg.GDP at 2.1 A resolution. It reveals that the G-domain fold and Mg.GDP binding site of Gem are similar to those found for other Ras family GTPases. The first part of the C-terminal extension adopts an alpha-helical conformation that extends along the alpha5 helix and interacts with the tip of the interswitch. Biochemical studies show that the affinities of Gem for GDP and GTP are considerably lower (micromolar range) compared with H-Ras, independent of the presence or absence of N- and C-terminal extensions, whereas its GTPase activity is higher than that of H-Ras and regulated by both extensions. We show how the bulky DXWEX motif, characteristic of the switch II of RGK proteins, affects the conformation of switch I and the phosphate-binding site. Altogether, our data reveal that Gem is a bona fide GTPase that exhibits striking structural and biochemical features that should impact its regulation and cellular activities. PMID- 17107950 TI - Lipid homeostasis and lipoprotein secretion in Niemann-Pick C1-deficient hepatocytes. AB - Niemann-Pick C (NPC) disease is a fatal inherited disorder characterized by an accumulation of cholesterol and other lipids in late endosomes/lysosomes. Although this disease is considered to be primarily a neurodegenerative disorder, many NPC patients suffer from liver disease. We have investigated alterations that occur in hepatic lipid homeostasis using primary hepatocytes isolated from NPC1-deficient mice. The cholesterol content of Npc1(-/-) hepatocytes was 5-fold higher than that of Npc1(+/+) hepatocytes; phospholipids and cholesteryl esters also accumulated. In contrast, the triacylglycerol content of Npc1(-/-) hepatocytes was 50% lower than of Npc1(+/+) hepatocytes. We hypothesized that the cholesterol sequestration induced by NPC1 deficiency might inhibit very low density lipoprotein secretion. However, this process was enhanced by NPC1 deficiency and the secreted particles were enriched in cholesteryl esters. We investigated the mechanisms responsible for these changes. The synthesis of phosphatidylcholine, cholesteryl esters, and cholesterol in hepatocytes was increased by NPC1 deficiency and the amount of the mature form of sterol response element-binding protein-1 was also increased. These observations indicate that the enhanced secretion of lipoproteins from NPC1-deficient hepatocytes is due, at least in part, to increased lipid synthesis. PMID- 17107949 TI - Ethanol-induced modulation of hepatocellular extracellular signal-regulated kinase-1/2 activity via 4-hydroxynonenal. AB - Modulation of the extracellular signal-regulated kinases (ERK-1/2), a signaling pathway directly associated with cell proliferation, survival, and homeostasis, has been implicated in several pathologies, including alcoholic liver disease. However, the underlying mechanism of ethanol-induced ERK-1/2 modulation remains unknown. This investigation explored the effects of ethanol-associated oxidative stress on constitutive hepatic ERK-1/2 activity and assessed the contribution of the lipid peroxidation product 4-hydroxynonenal (4-HNE) to the observations made in vivo. Constitutive ERK-1/2 phosphorylation was suppressed in hepatocytes isolated from rats chronically consuming ethanol for 45 days. This observation was associated with an increase in 4-HNE-ERK monomer adduct concentration and a hepatic cellular and lobular redistribution of ERK-1/2 that correlated with 4-HNE protein adduct accumulation. Chronic ethanol consumption was also associated with a decrease in hepatocyte nuclear ELK-1 phosphorylation, independent of changes in total nuclear ELK-1 protein. Primary hepatocytes treated with concentrations of 4 HNE consistent with those occurring during oxidative stress displayed a concentration-dependent decrease in constitutive ERK-1/2 phosphorylation, activity, and nuclear localization that negatively correlated with 4-HNE-ERK-1/2 monomer adduct accumulation. These data paralleled the decreased phosphorylation of the downstream kinase ELK-1. Molar ratios of purified ERK-2 to 4-HNE consistent with pathologic ratios found in vivo resulted in protein monomer adduct formation across a range of concentrations. Collectively, these data demonstrate a novel association between ethanol-induced lipid peroxidation and the inhibition of constitutive ERK-1/2, and suggest an inhibitory mechanism mediated by the lipid peroxidation product 4-hydroxynonenal. PMID- 17107951 TI - Characterization of the Plasmodium falciparum M17 leucyl aminopeptidase. A protease involved in amino acid regulation with potential for antimalarial drug development. AB - Amino acids generated from the catabolism of hemoglobin by intra-erythrocytic malaria parasites are not only essential for protein synthesis but also function in maintaining an osmotically stable environment, and creating a gradient by which amino acids that are rare or not present in hemoglobin are drawn into the parasite from host serum. We have proposed that a Plasmodium falciparum M17 leucyl aminopeptidase (PfLAP) generates and regulates the internal pool of free amino acids and therefore represents a target for novel antimalarial drugs. This enzyme has been expressed in insect cells as a functional 320-kDa homo-hexamer that is optimally active at neutral or alkaline pH, is dependent on metal ions for activity, and exhibits a substrate preference for N-terminally exposed hydrophobic amino acids, particularly leucine. PfLAP is produced by all stages in the intra-erythrocytic developmental cycle of malaria but was most highly expressed by trophozoites, a stage at which hemoglobin degradation and parasite protein synthesis are elevated. The enzyme was located by immunohistochemical methods and by transfecting malaria cells with a PfLAP-green fluorescent protein construct, to the cytosolic compartment of the cell at all developmental stages, including segregated merozoites. Amino acid dipeptide analogs, such as bestatin and its derivatives, are potent inhibitors of the protease and also block the growth of P. falciparum malaria parasites in culture. This study provides a biochemical basis for the antimalarial activity of aminopeptidase inhibitors. Availability of functionally active recombinant PfLAP, coupled with a simple enzymatic readout, will aid medicinal chemistry and/or high throughput approaches for the future design/discovery of new antimalarial drugs. PMID- 17107952 TI - Calcineurin regulates skeletal muscle metabolism via coordinated changes in gene expression. AB - The metabolic property of skeletal muscle adapts in response to an increased physiological demand by altering substrate utilization and gene expression. The calcium-regulated serine/threonine protein phosphatase calcineurin has been implicated in the transduction of motor neuron signals to alter gene expression programs in skeletal muscle. We utilized transgenic mice that overexpress an activated form of calcineurin in skeletal muscle (MCK-CnA*) to investigate the impact of calcineurin activation on metabolic properties of skeletal muscle. Activation of calcineurin increased glucose incorporation into glycogen and lipid oxidation in skeletal muscle. Activated calcineurin suppressed skeletal muscle glucose oxidation and increased lactate release. The enhancement in lipid oxidation was supported by increased expression of genes for lipid metabolism and mitochondrial oxidative phosphorylation. In a reciprocal fashion, several genes of glycolysis were down-regulated, whereas pyruvate dehydrogenase kinase 4 was markedly induced. This expression pattern was associated with decreased glucose utilization and enhanced glycogen storage. The peroxisome proliferator-activated receptors (PPARs) and PPARgamma coactivator 1alpha (PGC1alpha) are transcription regulators for the expression of metabolic and mitochondrial genes. Consistent with changes in the gene-regulatory program, calcineurin promoted the expression of PPARalpha, PPARdelta, and PPARgamma coactivator 1alpha in skeletal muscle. These results provide evidence that calcineurin-mediated skeletal muscle reprogramming induces the expression of several transcription regulators that coordinate changes in the expression of genes for lipid and glucose metabolism, which in turn alters energy substrate utilization in skeletal muscle. PMID- 17107953 TI - Urinary prostaglandin F2alpha is generated from the isoprostane pathway and not the cyclooxygenase in humans. AB - Prostaglandins (PGs) derived from the enzymatic oxidation of arachidonic acid by the cyclooxygenases (COXs) are potent lipid mediators involved in human physiology and pathophysiology. Structurally similar compounds, the isoprostanes (IsoPs), are generated from the free radical-catalyzed oxidation of arachidonic acid independent of COX. IsoPs exhibit significant bioactivity and play a role in the pathogenesis of diseases associated with oxidant injury. As one of the major PGs, prostaglandin F(2alpha) (PGF(2alpha)) is present in human urine in significant concentrations and is presumed to be derived from COX activity. We determined, however, that levels of putative PGF(2alpha) in urine cannot be suppressed by nonsteroidal anti-inflammatory agents, suggesting that it is generated via another mechanism(s). An important difference between COX-derived PGF(2alpha) and the IsoPs is that the former is an optically pure compound, whereas IsoPs are racemic. Utilizing a rodent model of oxidative stress, we now show that significant amounts of compounds identical in all respects to PGF(2alpha) and its enantiomer, ent-PGF(2alpha), are formed in equal amounts esterified in tissue phospholipids, suggesting that these compounds are derived via the IsoP pathway. Further, employing liquid chromatography/mass spectrometry, the vast majority of putative PGF(2alpha) in human urine is derived from the free radical-initiated peroxidation of arachidonate independent of COX and is composed of PGF(2alpha) and its enantiomer, although the latter compound is approximately 2-fold more abundant. Thus, quantification of urinary PGF(2alpha) actually reflects oxidative stress status as opposed to COX activity. Indeed, levels of this compound are elevated in urine from cigarette smokers and in humans with hypercholesterolemia, two conditions associated with oxidant stress. The elucidation that urinary PGF(2alpha) in humans is derived from the IsoP pathway has implications regarding PG formation and inhibition in vivo. PMID- 17107954 TI - Phosphorylation of the LFA-1 integrin beta2-chain on Thr-758 leads to adhesion, Rac-1/Cdc42 activation, and stimulation of CD69 expression in human T cells. AB - Phosphorylation of the leukocyte function-associated antigen-1 (LFA-1) integrin beta2-chain on Thr-758 occurs after T cell receptor stimulation and leads to 14-3 3 recruitment to the integrin, actin cytoskeleton reorganization, and increased adhesion. Here, we have investigated the signaling effects of beta2 integrin Thr 758 phosphorylation. A penetratin-coupled phospho-Thr-758-beta2 peptide (mimicking the part of the integrin beta-chain surrounding Thr-758) stimulated adhesion of human T cells to the LFA-1 ligand intercellular adhesion molecule-1 (ICAM-1). Additionally, the peptide activated the small GTPases Rac-1 and Cdc42 in T cells. Constitutively active forms of Rac-1 and Cdc42, but not Rho, could compensate for the reduction of cell adhesion to ICAM-1 caused by the T758A mutation in the beta2 integrin. Additionally, the active GTPases salvaged the cell-spreading defect of T758A integrin-transfected cells on coated ICAM-1. A dominant negative form of Cdc42, on the other hand, significantly reduced wild type beta2 integrin-mediated cell adhesion and spreading. In a T cell stimulation system, the pThr-758 penetratin peptide acted in a similar manner to coated ICAM 1 to increase T cell receptor-induced CD69 expression. These results show that Thr-758-phosphorylated LFA-1 is upstream of Rac-1/Cdc42, cell adhesion, and costimulatory activation of human T cells, thus identifying phosphorylation of Thr-758 in beta2 as a proximal element in LFA-1 signaling. PMID- 17107955 TI - Growth hormone corrects proliferation and transcription of phosphoenolpyruvate carboxykinase in livers of old mice via elimination of CCAAT/enhancer-binding protein alpha-Brm complex. AB - Growth hormone (GH), which is reduced with age, corrects the impaired proliferative capacity of livers of old animals. In this paper, we present a mechanism by which GH eliminates age-dependent negative control of proliferation and increases transcription of liver-specific genes in livers of old mice. The reduced proliferative capacities of the liver of old animals are associated with the CCAAT/enhancer-binding protein alpha (C/EBPalpha)-Brm complex, which inhibits E2F-dependent promoters. We found that a sequestration of C/EBPalpha into complexes with Brm leads to a weak interaction of C/EBPalpha with promoters of liver-specific genes, expression of which is reduced in old animals. Injection of either GH or the regulator of the amplitude of endogenous GH release, ghrelin, reduces the C/EBPalpha-Brm complex in livers of old mice, leading to a derepression of E2F targets, to increased interactions of C/EBPalpha with promoters of liver-specific genes, and to correction of their expression. GH dependent elimination of the complex is mediated by the inhibition of cyclin D3 CDK4 activity and by elevation of a phosphatase, protein phosphatase 2A, which dephosphorylates C/EBPalpha and dissociates the complex. PMID- 17107956 TI - The histone chaperone anti-silencing function 1 stimulates the acetylation of newly synthesized histone H3 in S-phase. AB - Anti-silencing function 1 (Asf1) is a highly conserved chaperone of histones H3/H4 that assembles or disassembles chromatin during transcription, replication, and repair. We have found that budding yeast lacking Asf1 has greatly reduced levels of histone H3 acetylated at lysine 9. Lysine 9 is acetylated on newly synthesized budding yeast histone H3 prior to its assembly onto newly replicated DNA. Accordingly, we found that the vast majority of H3 Lys-9 acetylation peaked in S-phase, and this S-phase peak of H3 lysine 9 acetylation was absent in yeast lacking Asf1. By contrast, deletion of ASF1 has no effect on the S-phase specific peak of H4 lysine 12 acetylation; another modification carried by newly synthesized histones prior to chromatin assembly. We show that Gcn5 is the histone acetyltransferase responsible for the S-phase-specific peak of H3 lysine 9 acetylation. Strikingly, overexpression of Asf1 leads to greatly increased levels of H3 on acetylation on lysine 56 and Gcn5-dependent acetylation on lysine 9. Analysis of a panel of Asf1 mutations that modulate the ability of Asf1 to bind to histones H3/H4 demonstrates that the histone binding activity of Asf1 is required for the acetylation of Lys-9 and Lys-56 on newly synthesized H3. These results demonstrate that Asf1 does not affect the stability of the newly synthesized histones per se, but instead histone binding by Asf1 promotes the efficient acetylation of specific residues of newly synthesized histone H3. PMID- 17107957 TI - R-Ras controls axon specification upstream of glycogen synthase kinase-3beta through integrin-linked kinase. AB - The initial event in establishing a polarized neuron is the specification of a single axon. Spatially regulated glycogen synthase kinase-3beta (GSK-3beta) activity is critical for specifying axon-dendrite fate; however, the upstream signaling of GSK-3beta in the determination of neuronal polarity still remains obscure. Here, we found that, in cultured hippocampal neurons, the small GTPase R Ras selectively localized in a single neurite of stage 2 neurons and that its activity increased after plating and peaked between stages 2 and 3. Ectopic expression of R-Ras induced global inactivation of GSK-3beta and formation of multiple axons, whereas knockdown of endogenous R-Ras by RNA interference blocked GSK-3beta inactivation and axon formation. GSK-3beta inactivation and axon formation by R-Ras required integrin-linked kinase (ILK), and subcellular localization of ILK was strictly regulated by R-Ras-mediated phosphatidylinositol 3-kinase activity. In addition, membrane targeting of ILK was sufficient to inactivate GSK-3beta and to form multiple axons. Our study demonstrates a novel role of R-Ras and ILK upstream of GSK-3beta in the regulation of neuronal polarity. PMID- 17107958 TI - Autoprocessing of Helicobacter pylori gamma-glutamyltranspeptidase leads to the formation of a threonine-threonine catalytic dyad. AB - Helicobacter pylorigamma-glutamyltranspeptidase (HpGT) is a glutathione-degrading enzyme that has been shown to be a virulence factor in infection. It is expressed as a 60-kDa inactive precursor that must undergo autocatalytic processing to generate a 40-kDa/20-kDa heterodimer with full gamma-glutamyl amide bond hydrolase activity. The new N terminus of the processed enzyme, Thr-380, is the catalytic nucleophile in both the autoprocessing and enzymatic reactions, indicating that HpGT is a member of the N-terminal nucleophile hydrolase superfamily. To further investigate activation as a result of autoprocessing, the structure of HpGT has been determined to a resolution of 1.9 A. The refined model contains two 40-kDa/20-kDa heterodimers in the asymmetric unit and has structural features comparable with other N-terminal nucleophile hydrolases. Autoprocessing of HpGT leads to a large conformational change, with the loop preceding the catalytic Thr-380 moving >35 A, thus relieving steric constraints that likely limit substrate binding. In addition, cleavage of the proenzyme results in the formation of a threonine-threonine dyad comprised of Thr-380 and a second conserved threonine residue, Thr-398. The hydroxyl group of Thr-398 is located equidistant from the alpha-amino group and hydroxyl side chain of Thr-380. Mutation of Thr-398 to an alanine results in an enzyme that is fully capable of autoprocessing but is devoid of enzymatic activity. Substrate docking studies in combination with homology modeling studies of the human homologue reveal additional mechanistic details of enzyme maturation and activation, substrate recognition, and catalysis. PMID- 17107959 TI - Comparative analysis of retroviral and native promoters driving expression of beta1,3-galactosyltransferase beta3Gal-T5 in human and mouse tissues. AB - Beta1,3-galactosyltransferase beta3Gal-T5 is highly expressed in the colons of humans and certain primates due to a retroviral long terminal repeat (LTR) acting as a strong promoter. Because this promoter is inactive in other human tissues or mice, we attempted to understand how adoption of a retrotransposon allowed the gene to acquire tissue-specific expression. We identified three novel 5'-UTRs of beta3Gal-T5 mRNA, types A, B, and C, and found widespread expression of the type A transcript at much lower levels than the LTR transcript, the expression of which is restricted to organs of the gastrointestinal tract. Expression of the type C 5'-UTR transcript was mostly restricted to the ileum, where it was expressed at high levels. We cloned the 5'-flanking regions of both types A and B 5'-UTRs, found deletion constructs functionally active as promoters, and identified CCAAT-binding factor (CBF) and hepatocyte nuclear factor 1 (HNF-1) as the principal nuclear factors controlling the promoters of types A and B 5'-UTR transcripts, respectively. The CCAAT-binding factor binding site and the entire downstream sequence driving the expression of type A transcripts in humans are structurally and functionally conserved in mice, where they constitute a uniquebeta3Gal-T5 promoter that appears to be the ancestral promoter of the gene. The HNF-1 binding motif of the second human promoter is identical to the HNF 1/Cdx binding motif of the LTR promoter but is in the antisense orientation, resulting in much lower binding affinity and promoter strength. These data may explain the successful insertion of the transposon during evolution. PMID- 17107960 TI - L2dtl is essential for cell survival and nuclear division in early mouse embryonic development. AB - l(2)dtl (lethal (2) denticleless), is an embryonic lethal homozygous mutation initially identified in Drosophila melanogaster that produces embryos that lack ventral denticle belts. In addition to nucleotide sequence, bioinformatic analysis has revealed a conservation of critical functional motifs among the human L2DTL, mouse L2dtl, and Drosophila l(2)dtl proteins. The function of the L2DTL protein in the development of mammalian embryos was studied using targeted disruption of the L2dtl gene in mice. The knock-out resulted in early embryonic lethality. L2dtl-/- embryos were deformed and terminated development at the 4-8 cell stage. Microinjection of a small interfering RNA (siRNA) vector (siRNA L2dtl) into the two-cell stage nuclei of wild-type mouse embryos led to cell cycle progression failure, termination of cell division, and, eventually, embryonic death during the preimplantation stage. Morphological studies of the embryos 54 h after injection showed fragmentation of mitotic chromosomes and chromosomal lagging, hallmarks of mitotic catastrophe. The siRNA-L2dtl-treated embryos eventually lysed and failed to develop into blastocysts after 72 h of in vitro culturing. However, the embryos developed normally after they were microinjected into one nucleus of the two-celled embryos. The siRNA studies in HeLa cells showed that L2dtl protein depletion results in multinucleation and down-regulation of phosphatidylinositol 3-kinase, proliferating cell nuclear antigen, and PTTG1/securin, which might partially explain the mitotic catastrophe observed in L2dtl-depleted mouse embryos. Based on these findings, we conclude that L2dtl gene expression is essential for very early mouse embryonic development. PMID- 17107961 TI - Metabolic diapause in pancreatic beta-cells expressing a gain-of-function mutant of the forkhead protein Foxo1. AB - Diabetes is associated with decreased pancreatic beta-cell function and mass. It is unclear whether diabetes treatment should aim at restoring beta-cell performance/mass or at inducing "beta-cell rest" to prevent further deterioration. The transcription factor Foxo1 protects beta-cells against oxidative stress induced by hyperglycemia and prevents beta-cell replication in insulin-resistant states. Here we show that these combined effects are associated with a concerted repression of genes involved in glycolysis, nitric-oxide synthesis, G protein-coupled receptor signaling, and ion transport. Conversely, Foxo1 increases expression of several neurotransmitter receptors and fails to regulate target genes predicted from Caenorhabditis elegans and Drosophila studies. Functional analyses show decreased glucose utilization and insulin secretion in beta-cells overexpressing Foxo1. We propose the definition of "metabolic diapause" for the changes induced by Foxo1 to protect beta-cells against oxidative stress. The data provide genetic underpinning for the concept of beta-cell rest as a treatment goal in diabetes. PMID- 17107963 TI - MEK-ERK-mediated phosphorylation of Mdm2 at Ser-166 in hepatocytes. Mdm2 is activated in response to inhibited Akt signaling. AB - Mdm2 inactivates the tumor suppressor p53 and Akt has been shown to be a major activator of Mdm2 in many cell types. We have investigated the regulation of Mdm2 in hepatocytes. We found that growth factor-induced Ser-166 phosphorylation of Mdm2 was inhibited by the MEK inhibitors U0126 and PD98059 in HepG2 cells and in a rat liver cell line, TRL 1215. Also, bile acids and oxidative stress induced phosphorylation of Mdm2 at Ser-166 by an apparently MEK-ERK-dependent mechanism. In contrast, Ser-166 phosphorylation of Mdm2 in lung cells was mediated by Akt. Further studies revealed that phosphatidylinositol 3-kinase inhibitors LY294002 and wortmannin induced phosphorylated ERK Tyr-204 and pMdm2 Ser-166 phosphorylations in hepatocytes in culture and in rat hepatocytes in vivo. In HepG2 cells, this effect was inhibited by U0126 and PD98059. LY294002 also reduced the level of pRaf Ser-259. Furthermore, we have shown that myr-Akt induced overexpression of pAkt suppressed the levels of pMdm2 Ser-166 in hepatocytes. These data indicate a reversed relationship between Akt and Mdm2 in hepatocytes and suggest that Akt is a negative regulator of Raf-MEK-ERK-Mdm2 in this cell type. Ser-166 phosphorylation of Mdm2 has been shown to increase its ubiquitin ligase activity and increase p53 degradation, and our data indicated an attenuated p53 response to DNA damage in hepatocytes exhibiting high levels of pMdm2 Ser-166. Taken together, our data indicate that Mdm2 phosphorylation is regulated via MEK-ERK in hepatocytes. This Mdm2 signaling might be important for the regeneration of hepatocytes after centrilobular cell death. PMID- 17107962 TI - Proteolytic processing of delta-like 1 by ADAM proteases. AB - Delta-like 1 (Dll1) is a mammalian ligand for Notch receptors. Interactions between Dll1 and Notch in trans activate the Notch pathway, whereas Dll1 binding to Notch in cis inhibits Notch signaling. Dll1 undergoes proteolytic processing in its extracellular domain by ADAM10. In this work we demonstrate that Dll1 represents a substrate for several other members of the ADAM family. In co transfected cells, Dll1 is constitutively cleaved by ADAM12, and the N-terminal fragment of Dll1 is released to medium. ADAM12-mediated cleavage of Dll1 is cell density-dependent, takes place in cis orientation, and does not require the presence of the cytoplasmic domain of ADAM12. Full-length Dll1, but not its N- or C-terminal proteolytic fragment, co-immunoprecipitates with ADAM12. By using a Notch reporter construct, we show that Dll1 processing by ADAM12 increases Notch signaling in a cell-autonomous manner. Furthermore, ADAM9 and ADAM17 have the ability to process Dll1. In contrast, ADAM15 does not cleave Dll1, although the two proteins still co-immunoprecipitate with each other. Asn-353 present in the catalytic motif of ADAM12 and other Dll1-processing ADAMs, but absent in ADAM15, is necessary for Dll1 cleavage. Dll1 cleavage is reduced in ADAM9/12/15(-/-) mouse embryonic fibroblasts (MEFs), suggesting that the endogenous ADAM9 and/or ADAM12 present in wild type MEFs contribute to Dll1 processing. Finally, the endogenous Dll1 present in primary mouse myoblasts undergoes cleavage in confluent, differentiating myoblast cultures, and this cleavage is decreased by ADAM12 small interfering RNAs. Our findings expand the role of ADAM proteins in the regulation of Notch signaling. PMID- 17107964 TI - Telmisartan attenuates chronic ciclosporin A nephrotoxicity in a pig model. AB - BACKGROUND: We have previously demonstrated renal enlargement in pigs treated with ciclosporin A (CsA) 10 mg/kg/day orally for 6 months. The aim of the study was to investigate the effect of oral CsA (10 mg/kg/day) for 12 months on kidney structure and function and the potential renoprotective role of angiotensin II (Ang II) receptor blocker telmisartan on chronic CsA nephrotoxicity in pigs. METHODS: Fourteen Gottingen minipigs aged 12-14 months were included: pigs received either CsA 10 mg/kg/day (n = 7) or CsA 10 mg/kg/day + telmisartan 40 mg/day (n = 7) orally for 12 months. At week 0, 12, 31, 38, 47 and 54, we measured body weight, mean arterial blood pressure (MAP), serum creatinine, whole blood trough CsA, plasma Ang II, haemoglobin and liver function parameters. Magnetic resonance imaging was used to estimate kidney length, volume, relative glomerular filtration rate (rGFR) and renal blood flow (RBF). Kidney tissue biopsies were used for conventional histological examination. RESULTS: Plasma Ang II levels were significantly higher during telmisartan treatment. Interstitial fibrosis and glomerulosclerosis occurred in both groups, but were attenuated in the telmisartan-treated pigs (P = 0.064). A significant increase in renal volume was seen in both groups, but tended to be lower in the CsA + telmisartan pigs at 54 weeks (P = 0.097). Telmisartan did not reduce MAP, RBF or rGFR. CONCLUSIONS: Long-term CsA treatment causes histopathological changes in the porcine kidney similar to those observed in humans and results in renal enlargement. Telmisartan attenuates the CsA-induced histopathological changes and enlargement in the pig kidney. PMID- 17107965 TI - Studies to identify the basis for an alkaline urine pH in patients with calcium hydrogen phosphate kidney stones. AB - BACKGROUND: Patients with CaHPO(4) kidney stones belong to a diagnostic category that has a high urine pH as its common feature. Our objective was to provide a new clinical approach to examine the basis for this high pH. METHODS: The study group consisted of 26 CaHPO(4) stone formers and 28 normal volunteers. Urine was collected q2h plus an overnight sample to identify patients with a urine pH > 6.5 for 12/24 h. Urine ammonium (U(NH4)), sulphate (U(SO4)) and citrate were measured and diet net alkali was calculated. RESULTS: Of the 26 patients, 13 had persistently alkaline urine. In 7/13, U(NH4) (68 +/- 13 mEq/day) and U(SO4) (57 +/- 7 mEq/day) were both high. In 6/13 patients, U(NH4) was the usual 31 +/- 3 mEq/day; in 4/6, U(NH4)/U(SO4) was 0.9 +/- 0.1; the cause of the alkaline urine pH seemed to be a dietary alkali load because the rise in urine pH was episodic and coincided with a high net diet alkali load and peak citrate excretion rates. The remaining two patients had a high U(NH4)/U(SO4) (2.2 and 1.6). Citrate excretion was very low in the male, but not in the female patient. CONCLUSIONS: There are heterogeneous causes for a persistently high urine pH. Two of the patients had a possible molecular basis: the lesion could be a low proximal convoluted tubule cell pH in the male and an increased entry of NH(3) into the late distal nephron in the female. PMID- 17107966 TI - Vitamin C deficiency in dialysis patients--are we perceiving the tip of an iceberg? PMID- 17107967 TI - Dynamic modelling and analysis of biochemical networks: mechanism-based models and model-based experiments. AB - Systems biology applies quantitative, mechanistic modelling to study genetic networks, signal transduction pathways and metabolic networks. Mathematical models of biochemical networks can look very different. An important reason is that the purpose and application of a model are essential for the selection of the best mathematical framework. Fundamental aspects of selecting an appropriate modelling framework and a strategy for model building are discussed. Concepts and methods from system and control theory provide a sound basis for the further development of improved and dedicated computational tools for systems biology. Identification of the network components and rate constants that are most critical to the output behaviour of the system is one of the major problems raised in systems biology. Current approaches and methods of parameter sensitivity analysis and parameter estimation are reviewed. It is shown how these methods can be applied in the design of model-based experiments which iteratively yield models that are decreasingly wrong and increasingly gain predictive power. PMID- 17107968 TI - The invasion promoting effect of microglia on glioblastoma cells is inhibited by cyclosporin A. AB - The invasion of tumour cells into brain tissue is a pathologic hallmark of WHO grades II-IV gliomas and contributes significantly to the failure of current therapeutic treatments. Activated microglial cells are abundant in brain tumours and may support tumour invasiveness. We have previously demonstrated that cyclosporin A (CsA) can affect growth of glioma cells in vitro by inhibiting signalling pathways, which are essential for tumour proliferation and invasiveness. In this work, we demonstrate that migration of EGFP-transfected glioblastoma cells in organotypic brain slices was significantly inhibited by treatment with CsA. On average 77% of untreated cells migrated beyond 500 mum, while only 28-33% cells migrated as far in the brain slices treated with CsA (P < 0.001). This inhibitory effect on glioblastoma invasion was lost when glioblastoma cells were injected into microglia-depleted brain slices. Moreover, CsA significantly inhibits intracranial glioma growth in vivo. We demonstrate that microglia-derived factors increase glioma invasiveness in Matrigel assay in vitro and this is associated with activation of the PI-3K/Akt signalling pathway. The invasion promoting effect of microglia is abolished in the presence of CsA. Furthermore, glioma-derived soluble factors induce morphological transformation of microglia and activate MAPK signalling, although production of pro inflammatory factors was not observed. Our findings that CsA interferes at clinically relevant concentrations with the tumour-promoting role of microglia and impairs invasive growth of glioma cells in vivo may provide a novel therapeutic strategy against gliomas. PMID- 17107969 TI - TISA: tissue-specific alternative splicing in human and mouse genes. AB - Alternative splicing (AS) is a mechanism by which multiple transcripts are produced from a single gene and is thought to be an important mechanism for tissue-specific expression of transcript isoforms. Here, we report a novel graphing method for transcript reconstruction and statistical prediction of tissue-specific AS. We applied three selection steps to generate the splice graph and predict the transcript isoforms: (i) a custom scoring rule for exon/intron sets, (ii) binomial statistics for selecting valid alternative splicing with a frequency of at least 1% for the predominant form and (iii) evaluation of transcript structure. We obtained 97 286 and 66 022 valid transcripts from 26 143 human and 27 741 mouse genes, respectively. In addition, we discovered 33 481 AS events for nine types of AS patterns in human. The statistical significance of tissue specificity for each gene, transcript and AS event was assessed based on EST tissue information, followed by a multiple testing correction procedure. In human, 12 711 genes, 16 016 transcripts and 1035 AS events were predicted to be tissue-specific (false discovery rate <0.01). This information on genes, transcript structures, AS events and their tissue specificities in human and mouse are freely accessible on the TISA website (http://tisa.kribb.re.kr/AGC/). PMID- 17107970 TI - Oral administration of itraconazole solution has superior efficacy in experimental oral and oesophageal candidiasis in mice than its intragastric administration. AB - OBJECTIVE: The therapeutic activities of cyclodextrin-associated itraconazole oral solution (itraconazole OS) by two administration routes in experimental oral and oesophageal candidiasis in mice were examined and compared. METHODS: Using experimental oral and oesophageal candidiasis models in ICR mice, we investigated the efficacy of oral and intragastric administration of itraconazole OS and checked the concentration of itraconazole and its metabolite hydroxyitraconazole (OH-itraconazole) in tongues or oesophagus tissue after administration of itraconazole OS. RESULTS: Oral administration of itraconazole OS at doses of 0.8, 4.0 or 20 mg/kg/day clearly decreased the number of viable Candida albicans cells in the oral cavity of mice with oral candidiasis in a dose-dependent manner at 3 days after infection. Intragastric administration of itraconazole OS at doses of 4 and 20 mg/kg/day once a day were also effective but to a lesser degree than that of oral administration. In the oesophageal candidiasis model, oral administration of itraconazole OS displayed superior therapeutic efficacy to the intragastric route. In coincidence with the greater efficacy, itraconazole was detected in lesional tissues after oral administration of itraconazole OS. CONCLUSIONS: Oral administration of itraconazole OS displayed therapeutic efficacy against murine oral and oesophageal candidiasis superior to that achieved by intragastric administration. This can be explained by there being higher concentrations of itraconazole in tongues or oesophagus tissues after administration of the suspension by the oral route. PMID- 17107971 TI - The five W's of progesterone receptors A and B: now we know where and when. PMID- 17107972 TI - Endocrine disruptors: do family lines carry an epigenetic record of previous generations' exposures? PMID- 17107974 TI - Safety of sperm washing and ART outcome in 741 HIV-1-serodiscordant couples. AB - BACKGROUND: To evaluate the safety of sperm washing and assisted reproduction technique (ART) outcome offered to serodiscordant couples with a human immunodeficiency virus-1 (HIV-1)-positive male. METHODS: Sperm washing was performed and checked by RT-PCR on each semen sample before its fresh usage. Intrauterine insemination (IUI) or IFV/ICSI was offered according to fertility profile of each couple. Non-infected women underwent HIV testing 2 weeks before each procedure and for up to 6 months after. RESULTS: Seven hundred and forty-one couples entered the study of a possible 2011 serodiscordant couples counselled over 4 years. Superovulation and IUI were performed in 581 couples, where the pregnancy rate per cycle and pregnancy rate per couple were 19 and 78%, respectively, with multiple pregnancy rate being 4%. One hundred and sixty couples were treated by IVF/ICSI, where pregnancy rate per cycle and per couple were 22 and 41%, respectively, with multiple pregnancy rate being 10%. All female partners were still HIV-1 negative at follow-up. CONCLUSION: Sperm washing within a programme of reproductive counselling was proved to be safe in this large series of serodiscordant couples. The overall pregnancy rate (70.3%), independent of the procedure used (IUI or IVF/ICSI), justifies the effort of the medical team in setting up and implementing dedicated centres and of the individual patient in seeking a safe pregnancy. PMID- 17107975 TI - Hypertension begets hypertrophy begets atrial fibrillation? Insights from yet another sheep model. PMID- 17107976 TI - Cause of death with bare metal and sirolimus-eluting stents. AB - AIMS: Although drug-eluting stents have assumed a dominant role in interventional cardiology, concern has been raised about the potential for long-term adverse outcomes, including death. The aim of the present study was to compare the incidence and cause of death between patients who received sirolimus-eluting or bare metal stents. METHODS AND RESULTS: An integrated analysis was performed on 1748 patients enrolled in four prospective double-blind trials that randomly assigned patients to receive either a sirolimus-eluting or a bare metal stent for treatment of a single de novo coronary stenosis. During a mean follow-up of 2.6+/ 0.6 years, 64 patients (3.7%) died. Total mortality was 3.2% among 870 bare metal stent patients and 4.1% among 878 sirolimus-eluting stent patients (P=0.37); there was no difference in cardiac mortality (1.4 vs. 1.3%; P=0.55) or causes of death between these two groups. The predominant cause of death was non-cardiac. Cardiac death was most frequently assigned owing to unwitnessed death. Death due to acute myocardial infarction, congestive heart failure, and stent thrombosis occurred infrequently. CONCLUSION: At a mean follow-up of 2.6 years in percutaneous coronary intervention patients, the predominant cause of death was non-cardiac. There was no significant difference in either the frequency or the cause of death with implantation of either sirolimus-eluting or bare metal stents. PMID- 17107977 TI - Increased mortality after implantation of first generation drug-eluting stents: seeing the smoke, where is the fire? PMID- 17107978 TI - Is there a role for multislice computed tomography in aortic stenosis? PMID- 17107979 TI - The meta-analysis: supportive or illuminating? PMID- 17107980 TI - Mitral valve repair for severe mitral regurgitation: the way forward? AB - Severe primary mitral regurgitation (MR) has a poor outcome if left uncorrected. Successful mitral valve repair has the unique potential to restore normal life expectancy and is superior to valve replacement. Despite this, mitral repair is performed relatively infrequently and many patients with potentially reparable valves have a replacement instead, subjecting them to unnecessary risk. Surgery in asymptomatic patients is a particularly difficult issue with some units advocating surgery irrespective of symptoms, based purely on the severity of regurgitation. This strategy cannot be widely adopted with the current patchy provision of high-quality valve repair surgery. Misplaced enthusiasm for early operation runs the risk of a failed repair and the hazards of a mechanical prosthesis. To ensure optimal treatment for patients with MR, cardiologists must be aware of the indications for valve repair and ensure that patients with potentially reparable valves are referred to surgeons with proven expertise, even if this means a shift from established practice. Surgical units need to promote subspecialization and rigorously audit their outcomes. There are currently no agreed standards for best practice in mitral valve repair and this is an area where professional societies may wish to take a role. PMID- 17107981 TI - African-American and Hispanic ethnicities, renal involvement and obesity predispose to hypertension in systemic lupus erythematosus: results from LUMINA, a multiethnic cohort (LUMINAXLV). AB - OBJECTIVE: To examine the predictors of the occurrence of hypertension in a large multiethnic US cohort. PATIENTS AND METHODS: There were 614 patients with systemic lupus erythematoses (SLE; > or = 4 American College of Rheumatology revised criteria) with < or = 5 years of disease duration at entry into the cohort (T0) and of Hispanic (Texan or Puerto Rican), African-American or Caucasian ethnicity. T0 variables were compared between patients who did and did not develop hypertension (blood pressure > or = 140/90 mm Hg on at least two occasions and/or the use of antihypertensive drugs) after T0. Significant and clinically relevant variables were then examined by a stepwise logistic regression model. RESULTS: A total of 379 patients without hypertension at T0 were included (patients who developed hypertension prior to SLE diagnosis (n = 126) or before T0 (n = 109) were excluded). Predictors of hypertension were African-American and Texan-Hispanic ethnicities, renal involvement and a higher body mass index. CONCLUSIONS: Traditional cardiovascular risk factors, disease related factors and ethnicity play a role in the occurrence of hypertension in patients with SLE. Controlling renal involvement and optimising body weight may prevent the occurrence of hypertension. PMID- 17107982 TI - Reproducibility and sensitivity to change of four scoring methods for the radiological assessment of osteoarthritis of the hand. AB - BACKGROUND: Osteoarthritis (OA) of the hand could be a relevant model to study the progression of OA in structure-modification trials. Various methods are proposed to assess hand OA and its progression radiologically. OBJECTIVE: To compare intra-reader and inter-reader precision and sensitivity to change of four radiological scoring methods proposed in hand OA. METHODS: 2 trained readers scored separately 105 pairs of radiographs (baseline; year 1), selected from patients enrolled in a randomised controlled trial, for inter-reader reliability and sensitivity to change. They scored twice 60 pairs among the 105 for cross sectional and longitudinal intra-reader reliability. Radiological hand OA assessment used: global, Kellgren-Lawrence (KL), Kallman and Verbruggen scoring methods. Inter- and intra-reader reliabilities were studied using intraclass coefficient (ICC) and the Bland-Altman method. Sensitivity to change was compared by calculating the standardised response means. RESULTS: Transversal intra-reader reproducibility ICCs ranged from 0.922 to 0.999. Verbruggen ranked the highest, followed by the KL and Kallman methods. Inter-rater reliability was higher for the Verbruggen scores, followed by the KL, global and Kallman scores (ICC 0.706 0.999). Longitudinal intra-reader reliability (baseline; year 1) was better using the Kallman and KL (ICC 0.986 and 0.990), followed by the Verbruggen (0.941) or global methods (0.939). Standardised response means ranged from 0.24 (KL) to 0.29 (Kallman). CONCLUSION: All four methods compared well with respect to reliabilities. However, the Verbruggen and Kallman methods performed better. The method most sensitive to change was the Kallman method, followed by Verbruggen and global scores. This study also suggests that structural changes could be detected in hand OA over a 1-year period. PMID- 17107983 TI - Efficacy of rituximab (anti-CD20) for refractory systemic lupus erythematosus involving the central nervous system. AB - AIM: Neuropsychiatric systemic lupus erythematosus (NPSLE) is a serious treatment resistant phenotype of systemic lupus erythematosus. A standard treatment for NPSLE is not available. This report describes the clinical and laboratory tests of 10 patients with NPSLE before and after rituximab treatment, including changes in lymphocyte phenotypes. METHODS: Rituximab was administered at different doses in 10 patients with refractory NPSLE, despite intensive treatment. RESULTS: Treatment with rituximab resulted in rapid improvement of central nervous system related manifestations, particularly acute confusional state. Rituximab also improved cognitive dysfunction, psychosis and seizure, and reduced the SLE Disease Activity Index Score at day 28 in all 10 patients. These effects lasted for >1 year in five patients. Flow cytometric analysis showed that rituximab down regulated CD40 and CD80 on B cells and CD40L, CD69 and inducible costimulator on CD4+ T cells. CONCLUSIONS: Rituximab rapidly improved refractory NPSLE, as evident by resolution of various clinical signs and symptoms and improvement of radiographic findings. The down regulation of functional molecules on B and T cells suggests that rituximab modulates the interaction of activated B and T cells through costimulatory molecules. These results warrant further analysis of rituximab as treatment for NPSLE. PMID- 17107984 TI - Impact of case management (Evercare) on frail elderly patients: controlled before and after analysis of quantitative outcome data. AB - OBJECTIVES: To determine the impact on outcomes in patients of the Evercare approach to case management of elderly people. DESIGN: Practice level before and after analysis of hospital admissions data with control group. SETTING: Nine primary care trusts in England that, in 2003-5, piloted case management of elderly people selected as being at high risk of emergency admission. MAIN OUTCOME MEASURES: Rates of emergency admission, emergency bed days, and mortality from April 2001 to March 2005 in 62 Evercare practices and 6960-7695 control practices in England (depending on the analysis being carried out). RESULTS: The intervention had no significant effect on rates of emergency admission (increase 16.5%, (95% confidence interval -5.7% to 38.7%), emergency bed days (increase 19.0%, -5.3% to 43.2%), and mortality (increase 34.4%, -1.7% to 70.3%) for a high risk population aged >65 with a history of two or more emergency admissions in the preceding 13 months. For the general population aged > or =65 effects on the rates of emergency admission (increase 2.5%, -2.1% to 7.0%), emergency bed days (decrease -4.9%, -10.8% to 1.0%), and mortality (increase 5.5%, -3.5% to 14.5%) were also non-significant. CONCLUSIONS: Case management of frail elderly people introduced an additional range of services into primary care without an associated reduction in hospital admissions. This may have been because of identification of additional cases. Employment of community matrons is now a key feature of case management policy in the NHS in England. Without more radical system redesign this policy is unlikely to reduce hospital admissions. PMID- 17107985 TI - Case management for elderly people in the community. PMID- 17107986 TI - BALF-derived fibroblasts differ from biopsy-derived fibroblasts in systemic sclerosis. AB - Growth of fibroblasts from bronchoalveolar lavage fluid (BALF) in patients with systemic sclerosis (SSc) has previously been described. The purpose of the present study was to characterise fibroblasts from BALF and bronchial biopsies from SSc patients with alveolitis and from controls, to analyse fibroblast proliferation, migration, stress fibres and proteoglycan production. BALF and bronchial biopsies were collected from 10 patients with SSc and alveolitis and from 15 controls. Outgrowth of fibroblasts was observed from the BALF of four patients, particularly in those with a markedly increased percentage of eosinophils in BALF, but not in any member of the control group. Increased levels of granulocyte-macrophage colony-stimulating factor, correlating with the percentage of eosinophils in BALF, were found in patients when compared with controls. Fibroblasts from BALF showed an elongated, mobile phenotype and increased proteoglycan production compared to the corresponding biopsy fibroblasts. In conclusion, outgrowth of fibroblasts with an altered phenotype is reported from bronchoalveolar lavage fluid in systemic sclerosis patients with alveolitis and an increased percentage of eosinophils in the bronchoalveolar lavage fluid. These findings indicate a possible role for eosinophil-fibroblast interaction in pulmonary fibrosis in systemic sclerosis. PMID- 17107987 TI - Effects of rehabilitation on chest wall volume regulation during exercise in COPD patients. AB - In order to investigate underlying mechanisms, the present authors studied the effect of pulmonary rehabilitation on the regulation of total chest wall and compartmental (ribcage, abdominal) volumes during exercise in patients with chronic obstructive pulmonary disease. In total, 20 patients (forced expiratory volume in one second, mean +/- SEM 39 +/- 3% predicted) undertook high-intensity exercise 3 days x week(-1) for 12 weeks. Before and after rehabilitation, the changes in chest wall (cw) volumes at the end of expiration (EEV) and inspiration (EIV) were computed by optoelectronic plethysmography during incremental exercise to the limit of tolerance (W(peak)). Rehabilitation significantly improved W(peak) (57+/-7 versus 47+/-5 W). In the post-rehabilitation period and at identical work rates, significant reductions were observed in minute ventilation (35.1+/-2.7 versus 38.4+/-2.7 L x min(-1)), breathing frequency (26+/-1 versus 29+/-1 breaths x min(-1)) and EEV(cw) and EIV(cw) (by 182+/-79 and 136+/-37 mL, respectively). Inspiratory reserve volume was significantly increased (by 148+/ 70 mL). Volume reductions were attributed to significant changes in abdominal EEV and EIV (by 163+/-59 and 125+/-27 mL, respectively). The improvement in W(peak) was similar in patients who progressively hyperinflated during exercise and those who did not (24 and 26%, respectively). In conclusion, pulmonary rehabilitation lowers chest wall volumes during exercise by decreasing the abdominal volumes. The improvement in exercise capacity following rehabilitation is independent of the pattern of exercise-induced dynamic hyperinflation. PMID- 17107988 TI - Beclomethasone/formoterol versus budesonide/formoterol combination therapy in asthma. AB - The present study was designed to compare the fixed combination of beclomethasone and formoterol in a hydrofluoroalkane Modulite (Chiesi Farmaceutici, Parma, Italy) pressurised metered-dose inhaler (pMDI), with a combination of budesonide and formoterol administered via a Turbuhaler (AstraZeneca, Lund, Sweden) dry powder inhaler (DPI). This was a phase III, multinational, multicentre, double blind, double-dummy, randomised, two-arm parallel groups, controlled study design. After a 2-week run-in period, 219 patients with moderate-to-severe asthma were randomised to a 12-week treatment with beclomethasone 200 microg plus formoterol 12 microg b.i.d. delivered via a pMDI or budesonide 400 microg plus formoterol 12 microg b.i.d. delivered via a DPI. The analysis of noninferiority on primary outcome, morning peak expiratory flow in the last 2 weeks of treatment, showed no difference between groups. A statistically significant improvement from baseline in lung function, symptoms and rescue medication use was observed in both groups at all time-points. No differences were observed between treatments in either rate of asthma exacerbations or frequency of adverse events. The new fixed combination of beclomethasone and formoterol in hydrofluoroalkane Modulite pressurised metered-dose inhaler is equivalent to the marketed combination of budesonide and formoterol in terms of efficacy and tolerability profile. PMID- 17107989 TI - Dendritic cell recruitment in lesions of human and experimental pulmonary hypertension. AB - In the present study, the hypothesis that dendritic cells (DCs), key players in immunity and tolerance, might be involved in the immunopathology of idiopathic pulmonary arterial hypertension (IPAH) was tested. The phenotype and localisation of DCs were characterised by immunohistochemistry and double-labelling immunofluorescence in lung samples from controls, human IPAH patients and an experimental pulmonary hypertension model (monocrotaline-exposed rats). As compared with controls, morphometric analysis demonstrated increased numbers of dendritic cell-specific intercellular adhesion molecule-grabbing nonintegrin (DC SIGN)-positive cells in muscular pulmonary arteries in IPAH and OX-62-positive DCs in monocrotaline-induced pulmonary hypertension. In human samples, the mean+/ SEM number of DC-SIGN-positive cells.artery(-1) of 100-300 microm diameter was 1.4+/-0.4 in controls versus 26.4+/-2.7 in IPAH. In rats, the number of OX-62 positive cells.artery(-1) of 50-150 microm diameter was 0.5+/-0.2 in controls, and 0.7+/-0.5, 3.1+/-0.5 and 8.4+/-0.6 at day 7, 14 and 28 after monocrotaline exposure, respectively. Human complex lesions of muscular pulmonary arteries showed transmural DC infiltration. Phenotyping revealed an immature DC profile in human and experimental pulmonary hypertension. The results support the concept that immature dendritic cells accumulate in remodelled pulmonary vessels and hence could be involved in the immunopathology of pulmonary hypertension. PMID- 17107990 TI - Inflammatory changes, recovery and recurrence at COPD exacerbation. AB - Chronic obstructive pulmonary disease (COPD) exacerbations are associated with increased airway and systemic inflammation, though relationships between exacerbation recovery, recurrent exacerbation and inflammation have not been previously reported. In the present study, inflammatory changes at COPD exacerbations were related to clinical nonrecovery and recurrent exacerbations within 50 days. Serum interleukin (IL)-6, C-reactive protein (CRP), sputum IL-6 and IL-8 were measured in 73 COPD patients when stable, at exacerbation and at 7, 14 and 35 days post-exacerbation. In 23% of patients, symptoms did not recover to baseline by day 35. These patients had persistently higher levels of serum CRP during the recovery period. A total of 22% of the patients who had recurrent exacerbations within 50 days had significantly higher levels of serum CRP at day 14, compared with those without recurrences: 8.8 mg.L(-1) versus 3.4 mg.L(-1). Frequent exacerbators had a smaller reduction in systemic inflammation between exacerbation onset and day 35 compared with infrequent exacerbators. Nonrecovery of symptoms at chronic obstructive pulmonary disease exacerbation is associated with persistently heightened systemic inflammation. The time course of systemic inflammation following exacerbation is different between frequent and infrequent exacerbators. A high serum C-reactive protein concentration 14 days after an index exacerbation may be used as a predictor of recurrent exacerbations within 50 days. PMID- 17107991 TI - The 6-min walking distance: long-term follow up in patients with COPD. AB - The 6-min walking distance (6MWD) test is used in clinical practice and research into patients with chronic obstructive pulmonary disease (COPD). However, little is known about natural long-term change in this parameter. The 6MWD was measured at baseline and then annually for 5 yrs in 294 patients with COPD and its annual rate of decline was determined. Forced expiratory volume in one second (FEV1) was also measured and the relationship between changes in both markers was explored. At baseline, the median 6MWD was 380 m (range 160-600 m). It declined by 19% (16 m.yr(-1)) over the 5 yrs compared with baseline in patients with American Thoracic Society/European Respiratory Society stage III COPD (FEV1 30-50% predicted) and by 26% (15 m.yr(-1)) in patients with stage IV COPD (FEV1 <30% pred). Over the 5-yr follow-up, the proportion of patients with a minimal clinically significant decline of 54 m increased with the severity of the disease. It was 24% in stage II, 45% in stage III, and 63% in stage IV disease. In contrast, the rate of decline of FEV1 was greater in patients with milder airflow obstruction and lesser in patients with lower absolute FEV1 values. In conclusion, the 6-min walking distance test provides increasingly useful information as the severity of chronic obstructive pulmonary disease increases. PMID- 17107992 TI - ACE I/D but not AGT (-6)A/G polymorphism is a risk factor for mortality in ARDS. AB - The intrapulmonary renin-angiotensin system via tissue concentration of angiotensin II or bradykinin may have multiple effects on pulmonary pathophysiology. Therefore, it was investigated whether the presence of the D allele of the angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphism or the A allele of angiotensinogen (AGT) promoter polymorphism ( 6)A/G are independent risk factors for 30-day survival in acute respiratory distress syndrome (ARDS) patients. In a prospective study, adults (Germans of Caucasian ethnicity) with ARDS (n = 84) were recruited from the current authors' intensive care unit and genotyped for the ACE I/D and the AGT (-6)A/G polymorphisms, as were 200 healthy Caucasian controls. Mortality was increased in the ACE DD genotype compared with the I allele, and the ACE I/D polymorphism was an independent prognostic factor for 30-day survival. Patients with a homozygous DD genotype were at highest risk for death (hazard ratio 5.7; 95% confidence interval 1.7-19.2) compared with the II genotype. In contrast, the AGT (-6)A/G polymorphism was neither associated with an increased risk for development of ARDS nor with outcome. In patients with acute respiratory distress syndrome, the angiotensin-converting enzyme insertion/deletion polymorphism but not the angiotensinogen (-6)A/G promoter polymorphism is an independent risk factor with a pronounced effect on 30-day survival. PMID- 17107993 TI - Moisture damage and childhood asthma: a population-based incident case-control study. AB - Most previous studies on the association between moisture damage and asthma have been cross-sectional and relied on self-reported exposure and health. The present authors studied the association by carrying out careful home inspections among new, clinically determined cases of asthma and controls. New cases of asthma aged 12-84 months (n = 121) were recruited prospectively and matched for year of birth, sex and living area with two randomly selected population controls (n = 241). Trained engineers visited all homes. Both cases and controls had lived >or=75% of their lifetime or the past 2 yrs in their current home. Risk of asthma increased with severity of moisture damage and presence of visible mould in the main living quarters but not in other areas of the house. Cases more often had damage in their bedroom. Associations were comparable for atopic and nonatopic asthma and for children aged >30 months or 1.4 for hoarseness, lost voice, sinusitis). General respiratory symptoms were significantly elevated compared with a Dutch population sample (OR ranged 1.4-7.2). An excess risk for respiratory symptoms indicative of asthma was observed in swimming pool employees. Aggravation of existing respiratory disease or interactions between irritants and allergen exposures are the most likely explanations for the observed associations. PMID- 17107996 TI - Pitx2 promotes development of splanchnic mesoderm-derived branchiomeric muscle. AB - Recent experiments, showing that both cranial paraxial and splanchnic mesoderm contribute to branchiomeric muscle and cardiac outflow tract (OFT) myocardium, revealed unexpected complexity in development of these muscle groups. The Pitx2 homeobox gene functions in both cranial paraxial mesoderm, to regulate eye muscle, and in splanchnic mesoderm to regulate OFT development. Here, we investigated Pitx2 in branchiomeric muscle. Pitx2 was expressed in branchial arch core mesoderm and both Pitx2 null and Pitx2 hypomorphic embryos had defective branchiomeric muscle. Lineage tracing with a Pitx2cre allele indicated that Pitx2 mutant descendents moved into the first branchial arch. However, markers of both undifferentiated core mesoderm and specified branchiomeric muscle were absent. Moreover, lineage tracing with a Myf5cre allele indicated that branchiomeric muscle specification and differentiation were defective in Pitx2 mutants. Conditional inactivation in mice and manipulation of Pitx2 expression in chick mandible cultures revealed an autonomous function in expansion and survival of branchial arch mesoderm. PMID- 17107997 TI - Steel factor controls midline cell death of primordial germ cells and is essential for their normal proliferation and migration. AB - During germ-cell migration in the mouse, the dynamics of embryo growth cause many germ cells to be left outside the range of chemoattractive signals from the gonad. At E10.5, movie analysis has shown that germ cells remaining in the midline no longer migrate directionally towards the genital ridges, but instead rapidly fragment and disappear. Extragonadal germ cell tumors of infancy, one of the most common neonatal tumors, are thought to arise from midline germ cells that failed to die. This paper addresses the mechanism of midline germ cell death in the mouse. We show that at E10.5, the rate of apoptosis is nearly four-times higher in midline germ cells than those more laterally. Gene expression profiling of purified germ cells suggests this is caused by activation of the intrinsic apoptotic pathway. We then show that germ cell apoptosis in the midline is activated by down-regulation of Steel factor (kit ligand) expression in the midline between E9.5 and E10.5. This is confirmed by the fact that removal of the intrinsic pro-apoptotic protein Bax rescues the germ-cell apoptosis seen in Steel null embryos. Two interesting things are revealed by this: first, germ-cell proliferation does not take place in these embryos after E9.0; second, migration of germ cells is highly abnormal. These data show first that changing expression of Steel factor is required for normal midline germ cell death, and second, that Steel factor is required for normal proliferation and migration of germ cells. PMID- 17107998 TI - Heparan sulfate biosynthetic gene Ndst1 is required for FGF signaling in early lens development. AB - Multiple signaling molecules, including bone morphogenic proteins (BMP) and fibroblast growth factors (FGF), play important roles in early lens development. However, how these morphogens are regulated is still largely unknown. Heparan sulfate participates in both morphogen transport and morphogen-receptor interaction. In this study, we demonstrate that inactivation of the heparan sulfate biosynthetic gene Ndst1 resulted in invagination defects of the early lens and in the disruption of lens-determination gene expression, leading to severe lens hypoplasia or anophthalmia. Ndst1 mutants exhibited reduced sulfation of heparan sulfate, but both BMP- and Wnt-signaling remained unchanged. Instead, these embryos showed diminished binding of a subset of FGF proteins to FGF receptors. Consistent with disruption of FGF signaling, expression of phospho-Erk and ERM were also downregulated in Ndst1-mutant lenses. Taken together, these results establish an important role of Ndst1 function in FGF signaling during lens development. PMID- 17107999 TI - Juxtaposed Polycomb complexes co-regulate vertebral identity. AB - Best known as epigenetic repressors of developmental Hox gene transcription, Polycomb complexes alter chromatin structure by means of post-translational modification of histone tails. Depending on the cellular context, Polycomb complexes of diverse composition and function exhibit cooperative interaction or hierarchical interdependency at target loci. The present study interrogated the genetic, biochemical and molecular interaction of BMI1 and EED, pivotal constituents of heterologous Polycomb complexes, in the regulation of vertebral identity during mouse development. Despite a significant overlap in dosage sensitive homeotic phenotypes and co-repression of a similar set of Hox genes, genetic analysis implicated eed and Bmi1 in parallel pathways, which converge at the level of Hox gene regulation. Whereas EED and BMI1 formed separate biochemical entities with EzH2 and Ring1B, respectively, in mid-gestation embryos, YY1 engaged in both Polycomb complexes. Strikingly, methylated lysine 27 of histone H3 (H3-K27), a mediator of Polycomb complex recruitment to target genes, stably associated with the EED complex during the maintenance phase of Hox gene repression. Juxtaposed EED and BMI1 complexes, along with YY1 and methylated H3-K27, were detected in upstream regulatory regions of Hoxc8 and Hoxa5. The combined data suggest a model wherein epigenetic and genetic elements cooperatively recruit and retain juxtaposed Polycomb complexes in mammalian Hox gene clusters toward co-regulation of vertebral identity. PMID- 17108000 TI - Sprinter: a novel transmembrane protein required for Wg secretion and signaling. AB - Wingless (Wg) is a secreted ligand that differentially activates gene expression in target tissues. It belongs to the Wnt family of secreted signaling molecules that regulate cell-to-cell interactions during development. Activation of Wg targets is dependent on the ligand concentration in the extracellular milieu; cellular mechanisms that govern the synthesis, delivery and receipt of Wg are elaborate and complex. We have identified sprinter (srt), which encodes a novel, evolutionarily conserved transmembrane protein required for the transmission of the Wg signal. Mutations in srt cause the accumulation of Wg in cells that express it, and retention of the ligand prevents activation of its target genes in signal-receiving cells. In the absence of Srt activity, levels of Wg targets (including Engrailed in embryos lacking maternal and zygotic srt, and Senseless and Achaete in wing discs) are reduced. Activation of Wg targets in the receiving cells does not require srt. Hence, the function of Srt is restricted to events occurring within the Wg-producing cells. We show that srt is not required for any aspect of Hedgehog (Hh) signal transduction, suggesting specificity of srt for the Wg pathway. We propose that srt encodes a protein required for Wg secretion that regulates maturation, membrane targeting or delivery of Wg. Loss of srt function in turn diminishes Wg-pathway activation in receiving cells. PMID- 17108001 TI - Tsix defective in splicing is competent to establish Xist silencing. AB - Dosage differences of X-linked genes between male and female mammals are compensated for by a mechanism known as X-inactivation, and the noncoding Xist gene plays a crucial role in this process. The expression of Xist is regulated in cis by its noncoding antisense gene, Tsix, whose transcripts (though a fraction of them stay unspliced), are processed like common proteincoding RNAs. It has been suggested that certain classes of sense-antisense pairs of RNA are causally involved in not only gene regulation but also higher order chromatin structure in various organisms. In fact, recent studies demonstrated that Tsix modulates Xist expression through modification of the chromatin structure. It is still unknown, however, whether the RNA product is important for the function of Tsix or whether the antisense transcription is sufficient. To obtain insight into this issue, we eliminated the splicing products of Tsix in the mouse and explored the effects of this elimination on Tsix-mediated Xist silencing. To our surprise, the Xist locus was stably repressed on the X carrying the splicing-defective Tsix allele. Moreover, the repressive chromatin configuration was properly established at the Xist locus. These unexpected results indicate that the splicing products are dispensable for Tsix-mediated Xist silencing. PMID- 17108002 TI - Direct control of neurogenesis by selector factors in the fly eye: regulation of atonal by Ey and So. AB - During eye development, the selector factors of the Eyeless/Pax6 or Retinal Determination (RD) network control specification of organ-type whereas the bHLH type proneural factor Atonal drives neurogenesis. Although significant progress has been made in dissecting the acquisition of ;eye identity' at the transcriptional level, the molecular mechanisms underlying the progression from neuronal progenitor to differentiating neuron remain unclear. A recently proposed model for the integration of organ specification and neurogenesis hypothesizes that atonal expression in the eye is RD-network-independent and that Eyeless works in parallel or downstream of atonal to modify the neurogenetic program. We show here that distinct cis-regulatory elements control atonal expression specifically in the eye and that the RD factors Eyeless and Sine oculis function as direct regulators. We find that these transcription factors interact in vitro and provide indirect evidence that this interaction may be required in vivo. The subordination of neurogenesis to the RD pathway in the eye provides a direct mechanism for the coordination of neurogenesis and tissue specification during sensory organ formation. PMID- 17108003 TI - Eph/ephrins and N-cadherin coordinate to control the pattern of sympathetic ganglia. AB - Previous studies have suggested that the segmental pattern of neural-crest derived sympathetic ganglia arises as a direct result of signals that restrict neural crest cell migratory streams through rostral somite halves. We recently showed that the spatiotemporal pattern of chick sympathetic ganglia formation is a two-phase process. Neural crest cells migrate laterally to the dorsal aorta, then surprisingly spread out in the longitudinal direction, before sorting into discrete ganglia. Here, we investigate the function of two families of molecules that are thought to regulate cell sorting and aggregation. By blocking Eph/ephrins or N-cadherin function, we measure changes in neural crest cell migratory behaviors that lead to alterations in sympathetic ganglia formation using a recently developed sagittal slice explant culture and 3D confocal time lapse imaging. Our results demonstrate that local inhibitory interactions within inter-ganglionic regions, mediated by Eph/ephrins, and adhesive cell-cell contacts at ganglia sites, mediated by N-cadherin, coordinate to sculpt discrete sympathetic ganglia. PMID- 17108004 TI - Novel binding partners of Ldb1 are required for haematopoietic development. AB - Ldb1, a ubiquitously expressed LIM domain binding protein, is essential in a number of tissues during development. It interacts with Gata1, Tal1, E2A and Lmo2 to form a transcription factor complex regulating late erythroid genes. We identify a number of novel Ldb1 interacting proteins in erythroleukaemic cells, in particular the repressor protein Eto-2 (and its family member Mtgr1), the cyclin-dependent kinase Cdk9, and the bridging factor Lmo4. MO-mediated knockdowns in zebrafish show these factors to be essential for definitive haematopoiesis. In accordance with the zebrafish results these factors are coexpressed in prehaematopoietic cells of the early mouse embryo, although we originally identified the complex in late erythroid cells. Based on the change in subcellullar localisation of Eto-2 we postulate that it plays a central role in the transition from the migration and expansion phase of the prehaematopoietic cells to the establishment of definitive haematopoietic stem cells. PMID- 17108005 TI - The role of speculum and bimanual examinations when evaluating attendees at a sexually transmitted diseases clinic. AB - BACKGROUND: With the advent of molecular techniques, self-collected specimens without a clinician's examination are often adequate to detect common genital infections. OBJECTIVE: To evaluate the additional information that speculum and bimanual examinations provides clinicians in the routine evaluation of genital infections among attendees of a sexually transmitted disease (STD) clinic. METHODS: Cross-sectional study from a database of all visit records to two STD clinics in Baltimore between 1996 and 2002. Women were stratified on the basis of reason for visit. Proportional and likelihood ratio estimates of the speculum examination in detecting clinically relevant cervicovaginal lesions (leading to a diagnosis of other infections or outside referral for further management) and bimanual examination in detecting abnormalities (leading to a diagnosis of pelvic inflammatory disease or referral) are presented. RESULTS: 15 918 of 21 703 records were included: 12 073 were symptomatic (SYM; discharge, rash, abdominal pain, dysuria, genital irritation or odour), 1676 were asymptomatic contacts of an infected partner (CON) and 2169 were asymptomatic and presented for checkup (ASYM). The median age was 26 years; 94% were black. 11.8% of SYM, 4.6% of CON and 3.9% of ASYM patients had clinically meaningful lesions detected on speculum examination. The bimanual examination detected clinically relevant abnormalities in 6.5% of SYM, 0.8% of CON and 0.6% of ASYM patients. CONCLUSION: Symptomatic women are most likely to benefit from speculum and bimanual examinations. However, their yield in evaluating asymptomatic women is low. Prospective studies are needed to determine whether eliminating speculum and bimanual examinations in a subset of women would offer an operational advantage without compromising patient safety. PMID- 17108006 TI - Who declines to give a name at a sexual health service? AB - OBJECTIVE: To characterise patients who decline to provide their surname at a public sexual health centre. METHODS: A case-control study of all patients first attending the Sydney Sexual Health Centre from 1998 to 2004, using proforma collected electronic data to compare patients who did not provide their surname with those who did. In addition, the frequencies of the 10 most common surnames in the Sydney telephone directory were compared with the frequency of those names in the patient database. RESULTS: Of 27 241 patients, 1350 (5%) declined to provide their surname. The most common surnames were also over-represented, suggesting that aliases remained pervasive among the centre's patients. Sex workers, married people and people requesting HIV, hepatitis or sexually transmissible infection (STI) screening were all more likely to decline to provide a surname. By contrast, patients with symptoms, patients who were referred with a prior STI diagnosis and patients with a new bacterial or non-HIV viral STI or were a known contact with STI were all significantly more likely to provide a surname. Among patients who declined to provide a surname, 20 tested HIV positive. CONCLUSION: The anonymous option did not seem to eliminate the use of aliases. Although limited, there seems to be a market for anonymous sexual health screening, particularly for the asymptomatic. PMID- 17108007 TI - Involvement of oxidants and AP-1 in angiotensin II-activated NFAT3 transcription factor. AB - Cardiomyocyte hypertrophy is associated with multiple pathophysiological cardiovascular conditions. Recent studies have substantiated the finding that oxidants may contribute to the development of cardiomyocyte hypertrophy. Activation of the nuclear factor of activated T cells-3 (NFAT3) transcription factor has been shown to result from endocrine inducers of cardiomyocyte hypertrophy such as angiotensin II (ANG II) and serves as an important molecular regulator of cardiomyocyte hypertrophy. In this study, we found that antioxidant enzyme catalase and antioxidants N-acetyl-l-cysteine, alpha-phenyl-N-tert butylnitrone, and lipoic acid prevent ANG II from activating NFAT3 promoter luciferase. H(2)O(2) induces a time- and dose-dependent activation of NFAT3 transcription factor. A dominant negative form of NFAT3 transcription factor inhibited H(2)O(2) from activating NFAT3 promoter. An inhibitor of ERKs, but not phosphoinositide 3-kinase or p38 MAPKs, blocked NFAT3 activation by H(2)O(2). The NFAT3 binding site in the promoters of most genes contains a weak activator protein-1 (AP-1) binding site adjacent to the core consensus NFAT binding sequence. ERK inhibitor PD98059 was found previously to inhibit AP-1 activation by H(2)O(2). Inactivation of AP-1 transcription factor by cotransfection of a dominant negative c-Jun, TAM67, prevented H(2)O(2) or ANG II from activating NFAT3 promoter. NFAT3 promoter containing the core NFAT cis-element without AP-1 binding site failed to show activation by H(2)O(2) treatment. Our data suggest that hypertrophy inducers ANG II and H(2)O(2) may activate NFAT3 in cardiomyocyte through an AP-1 transcription factor-dependent mechanism. PMID- 17108008 TI - Differential regulation of myofilament protein isoforms underlying the contractility changes in skeletal muscle unloading. AB - Weight-bearing skeletal muscles change phenotype in response to unloading. Using the hindlimb suspension rat model, we investigated the regulation of myofilament protein isoforms in correlation to contractility. Four weeks of continuous hindlimb unloading produced progressive atrophy and contractility changes in soleus but not extensor digitorum longus muscle. The unloaded soleus muscle also had decreased fatigue resistance. Along with the decrease of myosin heavy chain isoform I and IIa and increase of IIb and IIx, coordinated regulation of thin filament regulatory protein isoforms were observed: gamma- and beta-tropomyosin decreased and alpha-tropomyosin increased, resulting in an alpha/beta ratio similar to that in normal fast twitch skeletal muscle; troponin I and troponin T (TnT) both showed decrease in the slow isoform and increases in the fast isoform. The TnT isoform switching began after 7 days of unloading and TnI isoform showed detectable changes at 14 days while other protein isoform changes were not significant until 28 days of treatment. Correlating to the early changes in contractility, especially the resistance to fatigue, the early response of TnT isoform regulation may play a unique role in the adaptation of skeletal muscle to unloading. When the fast TnT gene expression was upregulated in the unloaded soleus muscle, alternative RNA splicing switched to produce more high molecular weight acidic isoforms, reflecting a potential compensation for the decrease of slow TnT that is critical to skeletal muscle function. The results demonstrate that differential regulation of TnT isoforms is a sensitive mechanism in muscle adaptation to functional demands. PMID- 17108009 TI - Enhanced cadmium-induced testicular necrosis and renal proximal tubule damage caused by gene-dose increase in a Slc39a8-transgenic mouse line. AB - Resistance to cadmium (Cd)-induced testicular necrosis is an autosomal recessive trait defined as the Cdm locus. Using positional cloning, we previously identified the Slc39a8 (encoding an apical-surface ZIP8 transporter protein) as the gene most likely responsible for the phenotype. In situ hybridization revealed that endothelial cells of the testis vasculature express high ZIP8 levels in two sensitive inbred mouse strains and negligible amounts in two resistant strains. In the present study, we isolated a 168.7-kb bacterial artificial chromosome (BAC), carrying only the Slc39a8 gene, from a Cd-sensitive 129/SvJ BAC library and generated BAC-transgenic mice. The BTZIP8-3 line, having three copies of the 129/SvJ Slc39a8 gene inserted into the Cd-resistant C57BL/6J genome (having its normal two copies of the Slc39a8 gene), showed tissue-specific ZIP8 mRNA expression similar to wild-type mice, mainly in lung, testis, and kidney. The approximately 2.5-fold greater expression paralleled the fact that the BTZIP8-3 line has five copies, whereas wild-type mice have two copies, of the Slc39a8 gene. The ZIP8 mRNA and protein localized especially to endothelial cells of the testis vasculature in BTZIP8-3 mice. Cd treatment reversed Cd resistance (seen in nontransgenic littermates) to Cd sensitivity in BTZIP8-3 mice; reversal of the testicular necrosis phenotype confirms that Slc39a8 is unequivocally the Cdm locus. ZIP8 also localized specifically to the apical surface of proximal tubule cells in the BTZIP8-3 kidney. Cd treatment caused acute renal failure and signs of proximal tubular damage in the BTZIP8-3 but not nontransgenic littermates. BTZIP8-3 mice should be a useful model for studying Cd-induced disease in kidney. PMID- 17108010 TI - Hypertonic induction of aquaporin-5: novel role of hypoxia-inducible factor 1alpha. AB - Aquaporin-5 (AQP5) is a water channel protein expressed on the apical surface of alveolar epithelial type I cells in distal rat lung, suggesting a role for AQP5 in regulating alveolar surface liquid tonicity and/or cell volume. We investigated the molecular mechanisms underlying hypertonic induction of AQP5 in primary rat alveolar epithelial cells (AEC). Steady-state levels of AQP5 mRNA and protein were increased by exposure to sorbitol (200 mM in culture fluid) for 24 h. The increase in AQP5 was not accompanied by changes in mRNA half-life. Transduction of mouse lung epithelial (MLE-15) cells and primary rat AEC with lentivirus vectors encoding AQP5-luciferase demonstrated transcriptional activation of the reporter by exposure to hypertonic sorbitol solution. Hybridization of proteins from sorbitol-treated cells to a transcription factor DNA array demonstrated induction of hypoxia-inducible factor-1alpha (HIF-1alpha) by hypertonicity, which was confirmed by quantitative RT-PCR. Cotransfections of AQP5-luciferase with HIF-1alpha and HIF-1beta expression plasmids in MLE-15 cells led to dose-dependent transcriptional enhancement, which was partially abrogated by mutagenesis of putative HIF-1alpha binding sites in the proximal AQP5 promoter. Importantly, hypertonic induction of AQP5 was significantly inhibited by preventing HIF-1alpha induction with small interfering RNA. Hypertonicity induced activation of a transiently transfected vascular endothelial growth factor (VEGF) hypoxia response element-driven luciferase construct and increased expression of endogenous VEGF. These results demonstrate that hypertonic induction of both AQP5 and VEGF is transcriptionally regulated and mediated, at least in part, by HIF-1alpha, suggesting a novel role for HIF-1alpha in modulating cellular adaptive responses to osmotic stress. PMID- 17108011 TI - Effect of the hydrophilicity of acrylic intraocular lens material and haptic angulation on anterior capsule opacification. AB - AIM: To evaluate the influence of hydrophilic and hydrophobic acrylic material and haptic angulation on anterior capsule opacification (ACO). METHODS: Prospective study on 53 patients with bilateral age-related cataract. Patients underwent standard cataract surgery by the same surgeon and randomly received a hydrophilic acrylic intraocular lens (IOL) in one eye and a hydrophobic acrylic IOL in the other eye. Forty five of these patients completed the one-year follow up. The following parameters were assessed: decentration, buttonholing, anterolenticular gap (ALG), ACO, outgrowth and refractive outcome. RESULTS: At the one-year follow-up, ACO was seen in 80% of the hydrophilic and 100% of the hydrophobic IOLs. ACO was more intense in the hydrophobic IOLs (p<0.001). Outgrowth was seen in 42% of the hydrophilic and 2% of the hydrophobic IOLs (p = 0.0003). No case of persisting ALG was seen in the hydrophobic IOLs, but in 42% of the hydrophilic IOLs. The refractive outcome was -0.29 (SD 0.56) dioptres for the hydrophilic and 0.003 (SD 0.44) dioptres for the hydrophobic IOLs (p<0.001). CONCLUSION: These results suggest that there is less ACO in hydrophilic acrylic than in hydrophobic acrylic IOLs. Although material properties might play a role, the angulated haptics of the hydrophilic IOLs exert an additional effect by the persisting ALG and a lack of contact between the IOL and the anterior capsule. PMID- 17108012 TI - Identification of ganglion cell neurites in human subretinal and epiretinal membranes. AB - AIM: To determine whether neural elements are present in subretinal and epiretinal proliferative vitreoretinopathy (PVR) membranes as well as in diabetic, fibrovascular membranes removed from patients during vitrectomy surgery. METHODS: Human subretinal and epiretinal membranes of varying durations were immunolabelled with different combinations of antibodies to glial fibrillary acidic protein, vimentin, neurofilament protein and laminin. RESULTS: Anti neurofilament-labelled neurites from presumptive ganglion cells were frequently found in epiretinal membranes and occasionally found in subretinal membranes. In addition, the neurites were only observed in regions that also contained glial processes. CONCLUSIONS: These data demonstrate that neuronal processes are commonly found in human peri-retinal cellular membranes similar to that demonstrated in animal models. These data also suggest that glial cells growing out of the neural retina form a permissive substrate for neurite growth and thus may hold clues to factors that support this growth. PMID- 17108013 TI - Predictive factors for short-term visual outcome after intravitreal triamcinolone acetonide injection for diabetic macular oedema: an optical coherence tomography study. AB - AIM: To evaluate the predictive factors for visual outcome after intravitreal triamcinolone acetonide injection to treat refractory diabetic macular oedema (DME). METHODS: A retrospective chart review of patients with DME who met the following inclusion criteria was performed: clinically significant diabetic macular oedema, receipt of a 4 mg/0.1 ml intravitreal triamcinolone acetonide injection and an optical coherence tomography (OCT) of the macula performed up to 10 days before injection. All patients received a full ophthalmic examination including best-corrected Snellen visual acuity (VA). The main outcome measure was the mean change in vision 3 months after injection. RESULTS: Data from 73 eyes of 59 patients were analysed. After a mean follow-up of 324 days, the mean change in vision was -0.075 logarithm of minimum angle of resolution (logMAR) units, with 27.3% improving > or =3 lines, 6.8% declining > or =3 lines and 60.2% remaining stable within 1 line of baseline vision. Statistical analysis was performed using multivariate generalised estimating equations on the basis of data from 52 eyes of 42 patients. Factors associated with an improvement in vision 3 months after injection were worse baseline VA (-0.27 logMAR units/unit increase in baseline VA, p = 0.002) and presence of subretinal fluid (-0.17 logMAR units, p = 0.06). The presence of cystoid macular oedema negatively affected the visual outcome (0.15 logMAR units, p = 0.03). In addition, the presence of an epiretinal membrane (ERM) was associated with less visual improvement. ERM modified the effect of baseline VA as demonstrated by a significant interaction between these two variables (0.34 logMAR units/unit increase in baseline VA, p = 0.04). CONCLUSIONS: OCT factors and baseline VA can be useful in predicting the outcomes of VA 3 months after intravitreal triamcinolone acetonide injection in patients with refractory DME. PMID- 17108014 TI - Detection of Treponema pallidum in the vitreous by PCR. AB - BACKGROUND: Ocular involvement of syphilis still poses a clinical challenge due to the chameleonic behaviour of the disease. As the serodiagnosis has significant limitations, the direct detection of Treponema pallidum (TP) in the vitreous represents a desirable diagnostic tool. METHODS: Real-time polymerase chain reaction (PCR) for the detection of TP was applied in diagnostic vitrectomies of two patients with acute chorioretinitis. Qualitative verification of TP by real time PCR and melting point analysis according to a modified protocol was ruled out. Patients underwent complete ophthalmological examination with fundus photographs, fluorescein angiography, serological examination, antibiotic treatment and follow-up. RESULTS: In two cases of acute chorioretinitis of unknown origin, real-time PCR of vitreous specimens of both patients provided evidence of TP and was 100% specific. Initial diagnosis of presumed viral retinitis was ruled out by PCR of vitreous specimen. Patients were treated with systemic antibiotics and showed prompt improvement in visual function and resolution of fundus lesions. CONCLUSIONS: With real-time PCR, detection of TP in the vitreous was possible and delivered a sensitive, quick and inexpensive answer to a disease rather difficult to assess. In cases of acute chorioretinitis, the use of PCR-based assays of vitreous specimens in the diagnostic evaluation of patients is advisable. Although syphilitic chorioretinitis is a rare disease, PCR should include search for TP, as diagnostic dilemmas prolong definitive treatment in a sight-threatening disease. PMID- 17108015 TI - Iris atrophy in patients with newly diagnosed multibacillary leprosy: at diagnosis, during and after completion of multidrug treatment. AB - AIM: To describe the prevalence and incidence of iris atrophy in patients with multibacillary (MB) leprosy. METHODS AND PATIENTS: Prospective longitudinal cohort study. 301 newly diagnosed patients with MB leprosy were followed up during the 2 years of treatment with multidrug therapy (MDT) and for a further 5 years with biannual ocular examinations. Incidence of iris atrophy was calculated as the number of patients with iris atrophy per person-year (PY) of follow-up among those who did not have iris atrophy at baseline. Stepwise multiple regression confirmed the presence of specific associations of demographic and clinical characteristics (p<0.05) with iris atrophy, detected by univariate analysis. RESULTS: Iris atrophy was present in 6 (2%) patients at enrolment. During MDT, with 445 PYs of follow-up, 9 patients developed iris atrophy (IR 0.02, 95% CI 0.01 to 0.04) that was associated with cataract (HR 15.13, 95% CI 3.71 to 61.79, p<0.001) and corneal opacities (HR 6.83, 95% CI 1.62 to 28.8, p = 0.009). After MDT, with 2005 PYs of follow-up, 60 patients developed iris atrophy (IR 0.03, 95% CI 0.023 to 0.039) that was associated with age (per decade; HR 1.40, 95% CI 1.10 to 1.78, p = 0.006), skin smear positivity (HR 3.50, 95% CI 1.33 to 9.24, p = 0.011), cataract (HR 3.66, 95% CI 1.85 to 7.25, p<0.001), keratic precipitates (HR 2.76, 95% CI 1.02 to 7.47, p = 0.046) and corneal opacity (HR 3.95, 95% CI 1.86 to 8.38, p<0.001). CONCLUSIONS: Iris atrophy continues to develop in 3% of patients with MB leprosy every year after they complete a 2-year course of MDT, and is associated with age, increasing loads of mycobacteria, subclinical inflammation, cataract and corneal opacity. PMID- 17108016 TI - Waiting times for cataract surgery in ten European countries: an analysis using data from the SHARE survey. AB - AIMS: To assess waiting times for cataract surgery and their acceptance in European countries, and to find explanatory, country-specific health indicators. METHODS: Using data from the survey of health, ageing and retirement in Europe (SHARE), waiting times for cataract surgery of 245 respondents in ten countries were analysed with the help of linear regression. The influence of four country specific health indicators on waiting times was studied by multiple linear regression. The influence of waiting time and country on the wish to have surgery performed earlier was determined through logistic regression. Additional information was obtained for each country from opinion leaders in the field of cataract surgery. RESULTS: Waiting times differed significantly (p<0.001) between the ten analysed European countries. The length of wait was significantly influenced by the total expenditure on health (p<0.01) but not by the other country specific health indicators. The wish to have surgery performed earlier was determined by the length of wait (p<0.001) but not by the country where surgery was performed. CONCLUSION: The length of wait is influenced by the total expenditure on health, but not by the rate of public expenditure on health, by the physician density or by the acute bed density. The wish to have surgery performed earlier depends on the length of wait for surgery and is not influenced by the country. PMID- 17108017 TI - Concordance with heart failure medications: what do patients think? PMID- 17108018 TI - Richard Wiseman and the medical practitioners of restoration London. AB - The case histories used to illustrate Richard Wiseman's Several Chirurgicall Treatises (1676) reveal not only pathological and therapeutic detail but much information about the range of occupational relationships between Wiseman and the many London physicians with whom he collaborated on cases (more than forty are identified in an appendix to this article). His interaction with the two physicians with whom he had most to do, Francis Prujean and Walter Needham, exemplifies the extremes of such relationships. With Prujean, an older man and a leading figure in the College of Physicians, a formal and hierarchical relationship was observed, in which distinct occupational lines were maintained. In the case of Needham, a younger physician whose devotion to anatomical study was shared by Wiseman, a friendship developed between the two men that carried over into practice and tended to break down occupational and intellectual distinctions, so that Needham sometimes involved himself in Wiseman's surgical practice, and Wiseman initiated medical research later reported by Needham to the Royal Society. PMID- 17108019 TI - Severe acute respiratory syndrome coronavirus infection of mice transgenic for the human Angiotensin-converting enzyme 2 virus receptor. AB - Animal models for severe acute respiratory syndrome (SARS) coronavirus infection of humans are needed to elucidate SARS pathogenesis and develop vaccines and antivirals. We developed transgenic mice expressing human angiotensin-converting enzyme 2, a functional receptor for the virus, under the regulation of a global promoter. A transgenic lineage, designated AC70, was among the best characterized against SARS coronavirus infection, showing weight loss and other clinical manifestations before reaching 100% mortality within 8 days after intranasal infection. High virus titers were detected in the lungs and brains of transgene positive (Tg+) mice on days 1 and 3 after infection. Inflammatory mediators were also detected in these tissues, coinciding with high levels of virus replication. Lower virus titers were also detected in other tissues, including blood. In contrast, infected transgene-negative (Tg-) mice survived without showing any clinical illness. Pathologic examination suggests that the extensive involvement of the central nervous system likely contributed to the death of Tg+ mice, even though viral pneumonia was present. Preliminary studies with mice of a second lineage, AC63, in which the transgene expression was considerably less abundant than that in the AC70 line, revealed that virus replication was largely restricted to the lungs but not the brain. Importantly, despite significant weight loss, infected Tg+ AC63 mice eventually recovered from the illness without any mortality. The severity of the disease that developed in these transgenic mice--AC70 in particular--makes these mouse models valuable not only for evaluating the efficacy of antivirals and vaccines, but also for studying SARS coronavirus pathogenesis. PMID- 17108020 TI - Precise identification of a human immunodeficiency virus type 1 antigen processing mutant. AB - Human immunodeficiency virus type 1 (HIV-1) evokes a strong immune response, but the virus persists. Polymorphisms within known antigenic sites result in loss of immune recognition and can be positively selected. Amino acid variation outside known HLA class I restricted epitopes can also enable immune escape by interfering with the processing of the optimal peptide antigen. However, the lack of precise rules dictating epitope generation and the enormous genetic diversity of HIV make prediction of processing mutants very difficult. Polymorphism E169D in HIV-1 reverse transcriptase (RT) is significantly associated with HLA-B*0702 in HIV-1-infected individuals. This polymorphism does not map within a known HLA B*0702 epitope; instead, it is located five residues downstream of a HLA-B*0702 restricted epitope SPAIFQSSM (SM9). Here we investigate the association between E169D and HLA-B*0702 for immune escape via the SM9 epitope. We show that this single amino acid variation prevents the immune recognition of the flanked SM9 epitope by cytotoxic T cells through lack of generation of the epitope, which is a result of aberrant proteasomal cleavage. The E169D polymorphism also maps within and abrogates the recognition of an HLA-A*03-restricted RT epitope MR9. This study highlights the potential for using known statistical associations as indicators for viral escape but also the complexity involved in interpreting the immunological consequences of amino acid changes in HIV sequences. PMID- 17108021 TI - Oncolytic virotherapy synergism with signaling inhibitors: Rapamycin increases myxoma virus tropism for human tumor cells. AB - Myxoma virus is a rabbit-specific poxvirus pathogen that also exhibits a unique tropism for human tumor cells and is dramatically oncolytic for human cancer xenografts. Most tumor cell lines tested are permissive for myxoma infection in a fashion intimately tied to the activation state of Akt kinase. A host range factor of myxoma virus, M-T5, directly interacts with Akt and mediates myxoma virus tumor cell tropism. mTOR is a regulator of cell growth and metabolism downstream of Akt and is specifically inhibited by rapamycin. We report that treatment of nonpermissive human tumor cell lines, which normally restrict myxoma virus replication, with rapamycin dramatically increased virus tropism and spread in vitro. This increased myxoma replication is concomitant with global effects on mTOR signaling, specifically, an increase in Akt kinase. In contrast to the effects on human cancer cells, rapamycin does not increase myxoma virus replication in rabbit cell lines or permissive human tumor cell lines with constitutively active Akt. This indicates that rapamycin increases the oncolytic capacity of myxoma virus for human cancer cells by reconfiguring the internal cell signaling environment to one that is optimal for productive virus replication and suggests the possibility of a potentially therapeutic synergism between kinase signaling inhibitors and oncolytic poxviruses for cancer treatment. PMID- 17108022 TI - Intraspecies host specificity of a single-stranded RNA virus infecting a marine photosynthetic protist is determined at the early steps of infection. AB - Viruses are extremely abundant in seawater and are believed to be significant pathogens to photosynthetic protists (microalgae). Recently, several novel RNA viruses were found to infect marine photosynthetic protists; one of them is HcRNAV, which infects Heterocapsa circularisquama (Dinophyceae). There are two distinct ecotypes of HcRNAV with complementary intraspecies host ranges. Nucleotide sequence comparison between them revealed remarkable differences in the coat protein coding gene resulting in a high frequency of amino acid substitutions. However, the detailed mechanism supporting this intraspecies host specificity is still unknown. In this study, virus inoculation experiments were conducted with compatible and incompatible host-virus combinations to investigate the mechanism determining intraspecies host specificity. Cells were infected by adding a virus suspension directly to a host culture or by transfecting viral RNA into host cells by particle bombardment. Virus propagation was monitored by Northern blot analysis with a negative-strand-specific RNA probe, transmission electron microscopy, and a cell lysis assay. With compatible host-virus combinations, propagation of infectious progeny occurred regardless of the inoculation method used. When incompatible combinations were used, direct addition of a virus suspension did not even result in viral RNA replication, while in host cells transfected with viral RNA, infective progeny virus particles with a host range encoded by the imported viral RNA were propagated. This indicates that the intraspecies host specificity of HcRNAV is determined by the upstream events of virus infection. This is the first report describing the reproductive steps of an RNA virus infecting a photosynthetic protist at the molecular level. PMID- 17108023 TI - The Old World and New World alphaviruses use different virus-specific proteins for induction of transcriptional shutoff. AB - Alphaviruses are widely distributed throughout the world. During the last few thousand years, the New World viruses, including Venezuelan equine encephalitis virus (VEEV) and eastern equine encephalitis virus (EEEV), evolved separately from those of the Old World, i.e., Sindbis virus (SINV) and Semliki Forest virus (SFV). Nevertheless, the results of our study indicate that both groups have developed the same characteristic: their replication efficiently interferes with cellular transcription and the cell response to virus replication. Transcriptional shutoff caused by at least two of the Old World alphaviruses, SINV and SFV, which belong to different serological complexes, depends on nsP2, but not on the capsid protein, functioning. Our data suggest that the New World alphaviruses VEEV and EEEV developed an alternative mechanism of transcription inhibition that is mainly determined by their capsid protein, but not by the nsP2. The ability of the VEEV capsid to inhibit cellular transcription appears to be controlled by the amino-terminal fragment of the protein, but not by its protease activity or by the positively charged RNA-binding domain. These data provide new insights into alphavirus evolution and present a plausible explanation for the particular recombination events that led to the formation of western equine encephalitis virus (WEEV) from SINV- and EEEV-like ancestors. The recombination allowed WEEV to acquire capsid protein functioning in transcription inhibition from EEEV-like virus. Identification of the new functions in the New World alphavirus-derived capsids opens an opportunity for developing new, safer alphavirus-based gene expression systems and designing new types of attenuated vaccine strains of VEEV and EEEV. PMID- 17108024 TI - Severe acute respiratory syndrome coronavirus open reading frame (ORF) 3b, ORF 6, and nucleocapsid proteins function as interferon antagonists. AB - The severe acute respiratory syndrome coronavirus (SARS-CoV) is highly pathogenic in humans, with a death rate near 10%. This high pathogenicity suggests that SARS CoV has developed mechanisms to overcome the host innate immune response. It has now been determined that SARS-CoV open reading frame (ORF) 3b, ORF 6, and N proteins antagonize interferon, a key component of the innate immune response. All three proteins inhibit the expression of beta interferon (IFN-beta), and further examination revealed that these SARS-CoV proteins inhibit a key protein necessary for the expression of IFN-beta, IRF-3. N protein dramatically inhibited expression from an NF-kappaB-responsive promoter. All three proteins were able to inhibit expression from an interferon-stimulated response element (ISRE) promoter after infection with Sendai virus, while only ORF 3b and ORF 6 proteins were able to inhibit expression from the ISRE promoter after treatment with interferon. This indicates that N protein inhibits only the synthesis of interferon, while ORF 3b and ORF 6 proteins inhibit both interferon synthesis and signaling. ORF 6 protein, but not ORF 3b or N protein, inhibited nuclear translocation but not phosphorylation of STAT1. Thus, it appears that these three interferon antagonists of SARS-CoV inhibit the interferon response by different mechanisms. PMID- 17108025 TI - N-linked glycosylation status of classical swine fever virus strain Brescia E2 glycoprotein influences virulence in swine. AB - E2 is one of the three envelope glycoproteins of classical swine fever virus (CSFV). Previous studies indicate that E2 is involved in several functions, including virus attachment and entry to target cells, production of antibodies, induction of protective immune response in swine, and virulence. Here, we have investigated the role of E2 glycosylation of the highly virulent CSFV strain Brescia in infection of the natural host. Seven putative glycosylation sites in E2 were modified by site-directed mutagenesis of a CSFV Brescia infectious clone (BICv). A panel of virus mutants was obtained and used to investigate whether the removal of putative glycosylation sites in the E2 glycoprotein would affect viral virulence/pathogenesis in swine. We observed that rescue of viable virus was completely impaired by removal of all putative glycosylation sites in E2 but restored when mutation N185A reverted to wild-type asparagine produced viable virus that was attenuated in swine. Single mutations of each of the E2 glycosylation sites showed that amino acid N116 (N1v virus) was responsible for BICv attenuation. N1v efficiently protected swine from challenge with virulent BICv at 3 and 28 days postinfection, suggesting that glycosylation of E2 could be modified for development of classical swine fever live attenuated vaccines. PMID- 17108026 TI - Targeted disruption of Kaposi's sarcoma-associated herpesvirus ORF57 in the viral genome is detrimental for the expression of ORF59, K8alpha, and K8.1 and the production of infectious virus. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) ORF57 regulates viral gene expression at the posttranscriptional level during viral lytic infection. To study its function in the context of the viral genome, we disrupted KSHV ORF57 in the KSHV genome by transposon-based mutagenesis. The insertion of the transposon into the ORF57 exon 2 region also interrupted the 3' untranslated region of KSHV ORF56, which overlaps with the ORF57 coding region. The disrupted viral genome, Bac36-Delta57, did not express ORF57, ORF59, K8alpha, K8.1, or a higher level of polyadenylated nuclear RNA after butyrate induction and could not be induced to produce infectious viruses in the presence of valproic acid, a histone deacetylase inhibitor and a novel KSHV lytic cycle inducer. The ectopic expression of ORF57 partially complemented the replication deficiency of the disrupted KSHV genome and the expression of the lytic gene ORF59. The induced production of infectious virus particles from the disrupted KSHV genome was also substantially restored by the simultaneous expression of both ORF57 and ORF56; complementation by ORF57 alone only partially restored the production of virus, and expression of ORF56 alone showed no effect. Altogether, our data indicate that in the context of the viral genome, KSHV ORF57 is essential for ORF59, K8alpha, and K8.1 expression and infectious virus production. PMID- 17108027 TI - Vaccinia virus A6L encodes a virion core protein required for formation of mature virion. AB - Vaccinia virus A6L is a previously uncharacterized gene that is conserved in all sequenced vertebrate poxviruses. Here, we constructed a recombinant vaccinia virus encoding A6 with an epitope tag and showed that A6 was expressed in infected cells after viral DNA replication and packaged in the core of the mature virion. Furthermore, we showed that A6 was essential for vaccinia virus replication by performing clustered charge-to-alanine mutagenesis on A6, which resulted in two vaccinia virus mutants (vA6L-mut1 and vA6L-mut2) that displayed a temperature-sensitive phenotype. At 31 degrees C, both mutants replicated efficiently; however, at 40 degrees C, vA6L-mut1 grew to a low titer, while vA6L mut2 failed to replicate. The A6 protein expressed by vA6L-mut2 exhibited temperature-dependent instability. At the nonpermissive temperature, vA6L-mut2 was normal at viral gene expression and viral factory formation, but it was defective for proteolytic processing of the precursors of several major virion proteins, a defect that is characteristic of a block in virion morphogenesis. Electron microscopy further showed that the morphogenesis of vA6L-mut2 was arrested before the formation of immature virion with nucleoid and mature virion. Taken together, our data show that A6 is a virion core protein that plays an essential role in virion morphogenesis. PMID- 17108028 TI - Structure-based identification of a major neutralizing site in an adenovirus hexon. AB - Virus-specific neutralizing antibodies present an obstacle to the effective use of adenovirus vectors for gene therapy and vaccination. The specific sites recognized by neutralizing antibodies have not been identified for any adenovirus, but they have been proposed to reside within the hexon, in small regions of the molecule that are exposed on the capsid surface and possess sequences that vary among serotypes. We have mapped the epitopes recognized by a panel of seven hexon-specific monoclonal antibodies that neutralize the chimpanzee adenovirus 68 (AdC68). Surface plasmon resonance experiments revealed that the antibodies compete for a single hexon binding site, and experiments with synthetic peptides indicated that this site resides within just one small surface loop. Mutations within this loop (but not in other surface loops) permitted virus to escape neutralization by all seven monoclonal antibodies and to resist neutralization by polyclonal antisera obtained from animals immunized against AdC68. These results indicate that a single small surface loop defines a major neutralization site for AdC68 hexon. PMID- 17108029 TI - Failure to open the blood-brain barrier and deliver immune effectors to central nervous system tissues leads to the lethal outcome of silver-haired bat rabies virus infection. AB - Rabies is a lethal disease caused by neurotropic viruses that are endemic in nature. When exposure to a potentially rabid animal is recognized, prompt administration of virus-neutralizing antibodies, together with active immunization, can prevent development of the disease. However, once the nonspecific clinical symptoms of rabies appear conventional postexposure treatment is unsuccessful. Over the last decade, rabies viruses associated with the silver-haired bat (SHBRV) have emerged as the leading cause of human deaths from rabies in the United States and Canada as a consequence of the fact that exposure to these viruses is often unnoticed. The need to treat SHBRV infection following the development of clinical rabies has lead us to investigate why the immune response to SHBRV fails to protect at a certain stage of infection. We have established that measurements of innate and adaptive immunity are indistinguishable between mice infected with the highly lethal SHBRV and mice infected with an attenuated laboratory rabies virus strain. While a fully functional immune response to SHBRV develops in the periphery of infected animals, the invasion of central nervous system (CNS) tissues by immune cells is reduced and, consequently, the virus is not cleared. Our data indicate that the specific deficit in the SHBRV-infected animal is an inability to enhance blood brain barrier permeability in the cerebellum and deliver immune effectors to the CNS tissues. Conceivably, at the stage of infection where immune access to the infected CNS tissues is limited, either the provision or the development of antiviral immunity will be ineffective. PMID- 17108030 TI - A severe acute respiratory syndrome coronavirus that lacks the E gene is attenuated in vitro and in vivo. AB - A deletion mutant of severe acute respiratory syndrome coronavirus (SARS-CoV) has been engineered by deleting the structural E gene in an infectious cDNA clone that was constructed as a bacterial artificial chromosome (BAC). The recombinant virus lacking the E gene (rSARS-CoV-DeltaE) was rescued in Vero E6 cells. The recovered deletion mutant grew in Vero E6, Huh-7, and CaCo-2 cells to titers 20-, 200-, and 200-fold lower than the recombinant wild-type virus, respectively, indicating that although the E protein has an effect on growth, it is not essential for virus replication. No differences in virion stability under a wide range of pH and temperature were detected between the deletion mutant and recombinant wild-type viruses. Although both viruses showed the same morphology by electron microscopy, the process of morphogenesis seemed to be less efficient with the defective virus than with the recombinant wild-type one. The rSARS-CoV DeltaE virus replicated to titers 100- to 1,000-fold lower than the recombinant wild-type virus in the upper and lower respiratory tract of hamsters, and the lower viral load was accompanied by less inflammation in the lungs of hamsters infected with rSARS-CoV-DeltaE virus than with the recombinant wild-type virus. Therefore, the SARS-CoV that lacks the E gene is attenuated in hamsters, might be a safer research tool, and may be a good candidate for the development of a live attenuated SARS-CoV vaccine. PMID- 17108031 TI - E6AP ubiquitin ligase mediates ubiquitylation and degradation of hepatitis C virus core protein. AB - Hepatitis C virus (HCV) core protein is a major component of viral nucleocapsid and a multifunctional protein involved in viral pathogenesis and hepatocarcinogenesis. We previously showed that the HCV core protein is degraded through the ubiquitin-proteasome pathway. However, the molecular machinery for core ubiquitylation is unknown. Using tandem affinity purification, we identified the ubiquitin ligase E6AP as an HCV core-binding protein. E6AP was found to bind to the core protein in vitro and in vivo and promote its degradation in hepatic and nonhepatic cells. Knockdown of endogenous E6AP by RNA interference increased the HCV core protein level. In vitro and in vivo ubiquitylation assays showed that E6AP promotes ubiquitylation of the core protein. Exogenous expression of E6AP decreased intracellular core protein levels and supernatant HCV infectivity titers in the HCV JFH1-infected Huh-7 cells. Furthermore, knockdown of endogenous E6AP by RNA interference increased intracellular core protein levels and supernatant HCV infectivity titers in the HCV JFH1-infected cells. Taken together, our results provide evidence that E6AP mediates ubiquitylation and degradation of HCV core protein. We propose that the E6AP-mediated ubiquitin proteasome pathway may affect the production of HCV particles through controlling the amounts of viral nucleocapsid protein. PMID- 17108032 TI - Retinoic acid-inducible gene I mediates early antiviral response and Toll-like receptor 3 expression in respiratory syncytial virus-infected airway epithelial cells. AB - Respiratory syncytial virus (RSV) is one of the most common viral pathogens causing severe lower respiratory tract infections in infants and young children. Infected host cells detect and respond to RNA viruses using different mechanisms in a cell-type-specific manner, including retinoic acid-inducible gene I (RIG-I) dependent and Toll-like receptor (TLR)-dependent pathways. Because the relative contributions of these two pathways in the recognition of RSV infection are unknown, we examined their roles in this study. We found that RIG-I helicase binds RSV transcripts within 12 h of infection. Short interfering RNA (siRNA) mediated RIG-I "knockdown" significantly inhibited early nuclear factor-kappaB (NF-kappaB) and interferon response factor 3 (IRF3) activation 9 h postinfection (p.i.). Consistent with this finding, RSV-induced beta interferon (IFN-beta), interferon-inducible protein 10 (IP-10), chemokine ligand 5 (CCL-5), and IFN stimulated gene 15 (ISG15) expression levels were decreased in RIG-I-silenced cells during the early phase of infection but not at later times (18 h p.i.). In contrast, siRNA-mediated TLR3 knockdown did not affect RSV-induced NF-kappaB binding but did inhibit IFN-beta, IP-10, CCL-5, and ISG15 expression at late times of infection. Further studies revealed that TLR3 knockdown significantly reduced NF-kappaB/RelA transcription by its ability to block the activating phosphorylation of NF-kappaB/RelA at serine residue 276. We further found that TLR3 induction following RSV infection was regulated by RIG-I-dependent IFN-beta secreted from infected airway epithelial cells and was mediated by both IFN response-stimulated element (ISRE) and signal transducer and activator of transcription (STAT) sites in its proximal promoter. Together these findings indicate distinct temporal roles of RIG-I and TLR3 in mediating RSV-induced innate immune responses, which are coupled to distinct pathways controlling NF kappaB activation. PMID- 17108033 TI - Vaccinia virus interactions with the cell membrane studied by new chromatic vesicle and cell sensor assays. AB - The potential danger of cross-species viral infection points to the significance of understanding the contributions of nonspecific membrane interactions with the viral envelope compared to receptor-mediated uptake as a factor in virus internalization and infection. We present a detailed investigation of the interactions of vaccinia virus particles with lipid bilayers and with epithelial cell membranes using newly developed chromatic biomimetic membrane assays. This analytical platform comprises vesicular particles containing lipids interspersed within reporter polymer units that emit intense fluorescence following viral interactions with the lipid domains. The chromatic vesicles were employed as membrane models in cell-free solutions and were also incorporated into the membranes of epithelial cells, thereby functioning as localized membrane sensors on the cell surface. These experiments provide important insight into membrane interactions with and fusion of virions and the kinetic profiles of these processes. In particular, the data emphasize the significance of cholesterol/sphingomyelin domains (lipid rafts) as a crucial factor promoting bilayer insertion of the viral particles. Our analysis of virus interactions with polymer-labeled living cells exposed the significant role of the epidermal growth factor receptor in vaccinia virus infectivity; however, the data also demonstrated the existence of additional non-receptor-mediated mechanisms contributing to attachment of the virus to the cell surface and its internalization. PMID- 17108034 TI - The ERK mitogen-activated protein kinase pathway contributes to Ebola virus glycoprotein-induced cytotoxicity. AB - Ebola virus is a highly lethal pathogen that causes hemorrhagic fever in humans and nonhuman primates. Among the seven known viral gene products, the envelope glycoprotein (GP) alone induces cell rounding and detachment that ultimately leads to cell death. Cellular cytoxicity is not seen with comparable levels of expression of a mutant form of GP lacking a mucin-like domain (GPDeltamuc). GP induced cell death is nonapoptotic and is preceded by downmodulation of cell surface molecules involved in signaling pathways, including certain integrins and epidermal growth factor receptor. To investigate the mechanism of GP-induced cellular toxicity, we analyzed the activation of several signal transduction pathways involved in cell growth and survival. The active form of extracellular signal-regulated kinases types 1 and 2 (ERK1/2), phospho-ERK1/2, was reduced in cells expressing GP compared to those expressing GPDeltamuc as determined by flow cytometry, in contrast to the case for several other signaling proteins. Subsequent analysis of the activation states and kinase activities of related kinases revealed a more pronounced effect on the ERK2 kinase isoform. Disruption of ERK2 activity by a dominant negative ERK or by small interfering RNA-mediated ERK2 knockdown potentiated the decrease in alphaV integrin expression associated with toxicity. Conversely, activation of the pathway through the expression of a constitutively active form of ERK2 significantly protected against this effect. These results indicate that the ERK signaling cascade mediates GP-mediated cytotoxicity and plays a role in pathogenicity induced by this gene product. PMID- 17108035 TI - Cholesterol-depleting statin drugs protect postmitotically differentiated human neurons against ethanol- and human immunodeficiency virus type 1-induced oxidative stress in vitro. AB - The majority of human immunodeficiency virus type 1 (HIV-1)-infected individuals are either alcoholics or prone to alcoholism. Upon ingestion, alcohol is easily distributed into the various compartments of the body, particularly the brain, by crossing through the blood-brain barrier. Both HIV-1 and alcohol induce oxidative stress, which is considered a precursor for cytotoxic responses. Several reports have suggested that statins exert antioxidant as well as anti-inflammatory pleiotropic effects, besides their inherent cholesterol-depleting potentials. In our studies, postmitotically differentiated neurons were cocultured with HIV-1 infected monocytes, T cells, or their cellular supernatants in the presence of physiological concentrations of alcohol for 72 h. Parallel cultures were pretreated with statins (atorvastatin and simvastatin) with the appropriate controls, i.e., postmitotically differentiated neurons cocultured with uninfected cells and similar cultures treated with alcohol. The oxidative stress responses in the presence/absence of alcohol in these cultures were determined by the production of the well-characterized oxidative stress markers, 8-isoprostane-F2 alpha, total nitrates as an indicator for various isoforms of nitric oxide synthase activity, and heat shock protein 70 (Hsp70). An in vitro culture of postmitotically differentiated neurons with HIV-1-infected monocytes or T cells as well as supernatants from these cells enhanced the release of 8-isoprostane-F2 alpha in the conditioned medium six- to sevenfold (monocytes) and four- to fivefold (T cells). It was also observed that coculturing of HIV-1-infected primary monocytes over a time period of 72 h significantly elevated the release of Hsp70 compared with that of uninfected controls. Cellular supernatants of HIV 1-infected monocytes or T cells slightly increased Hsp70 levels compared to neurons cultured with uninfected monocytes or T-cell supernatants (controls). Ethanol (EtOH) presence further elevated Hsp70 in both infected and uninfected cultures. The amount of total nitrates was significantly elevated in the coculture system when both infected cells and EtOH were present. Surprisingly, pretreatment of postmitotic neurons with clinically available inhibitors of HMG coenzyme A reductase (statins) inhibited HIV-1-induced release of stress/toxicity associated parameters, i.e., Hsp70, isoprostanes, and total nitrates from HIV-1 infected cells. The results of this study provide new insights into HIV-1 neuropathogenesis aimed at the development of future HIV-1 therapeutics to eradicate viral reservoirs from the brain. PMID- 17108036 TI - Selective repression of translation by the brome mosaic virus 1a RNA replication protein. AB - Differential expression of viral replication proteins is essential for successful infection. We report here that overexpression of the brome mosaic virus (BMV) 1a protein can repress viral RNA replication in a dosage-dependent manner. Using RNA replication-incompetent reporter constructs, repression of translation from BMV RNA1 and RNA2 was observed, suggesting that the effect on translation of the BMV RNA replication proteins is responsible for the decrease in RNA levels. Furthermore, repression of translation by 1a required the B box in the 5' untranslated region (5' UTR); BMV RNA3 that lacks a B box in its 5' UTR is not subject to 1a-mediated translational inhibition. Mutations in either the methyltransferase or the helicase-like domains of 1a reduced the repression of replication and translation. These results suggest that in addition to its known functions in BMV RNA synthesis, 1a also regulates viral gene expression. PMID- 17108037 TI - Variable deficiencies in the interferon response enhance susceptibility to vesicular stomatitis virus oncolytic actions in glioblastoma cells but not in normal human glial cells. AB - With little improvement in the poor prognosis for humans with high-grade glioma brain tumors, alternative therapeutic strategies are needed. As such, selective replication-competent oncolytic viruses may be useful as a potential treatment modality. Here we test the hypothesis that defects in the interferon (IFN) pathway could be exploited to enhance the selective oncolytic profile of vesicular stomatitis virus (VSV) in glioblastoma cells. Two green fluorescent protein-expressing VSV strains, recombinant VSV and the glioma-adapted recombinant VSV-rp30a, were used to study infection of a variety of human glioblastoma cell lines compared to a panel of control cells, including normal human astrocytes, oligodendrocyte precursor cells, and primary explant cultures from human brain tissue. Infection rate, cell viability, viral replication, and IFN-alpha/beta-related gene expression were compared in the absence and presence of IFN-alpha or polyriboinosinic polyribocytidylic acid [poly(I:C)], a synthetic inducer of the IFN-alpha/beta pathway. Both VSV strains caused rapid and total infection and death of all tumor cell lines tested. To a lesser degree, normal cells were also subject to VSV infection. In contrast, IFN-alpha or poly(I:C) completely attenuated the infection of all primary control brain cells, whereas most glioblastoma cell lines treated with IFN-alpha or poly(I:C) showed little or no sign of protection and were killed by VSV. Together, our results demonstrate that activation of the interferon pathway protects normal human brain cells from VSV infection while maintaining the vulnerability of human glioblastoma cells to viral destruction. PMID- 17108038 TI - Latent protein LANA2 from Kaposi's sarcoma-associated herpesvirus interacts with 14-3-3 proteins and inhibits FOXO3a transcription factor. AB - The Kaposi's sarcoma-associated herpesvirus latent protein LANA2 has been suggested to have an important role in the transforming activity of the virus based on its capacity to inhibit p53 and PKR-dependent apoptosis as well as the interferon-dependent response. Here, we describe a novel interaction between LANA2 and both the phosphoserine/phosphothreonine-binding 14-3-3 proteins and the transcription factor FOXO3a. In addition, our results indicate that LANA2 inhibits the transcriptional activity of FOXO3a and blocks the G2/M arrest induced by 14-3-3 protein overexpression. These results suggest a novel mechanism by which LANA2 may promote tumorigenesis. PMID- 17108039 TI - Inhibition of heavy chain and beta2-microglobulin synthesis as a mechanism of major histocompatibility complex class I downregulation during Epstein-Barr virus replication. AB - The mechanisms of major histocompatibility complex (MHC) class I downregulation during Epstein-Barr virus (EBV) replication are not well characterized. Here we show that in several cell lines infected with a recombinant EBV strain encoding green fluorescent protein (GFP), the virus lytic cycle coincides with GFP expression, which thus can be used as a marker of virus replication. EBV replication resulted in downregulation of MHC class II and all classical MHC class I alleles independently of viral DNA synthesis or late gene expression. Although assembled MHC class I complexes, the total pool of heavy chains, and beta2-microglobulin (beta2m) were significantly downregulated, free class I heavy chains were stabilized at the surface of cells replicating EBV. Calnexin expression was increased in GFP+ cells, and calnexin and calreticulin accumulated at the cell surface that could contribute to the stabilization of class I heavy chains. Decreased expression levels of another chaperone, ERp57, and TAP2, a transporter associated with antigen processing and presentation, correlated with delayed kinetics of MHC class I maturation. Levels of both class I heavy chain and beta2m mRNA were reduced, and metabolic labeling experiments demonstrated a very low rate of class I heavy chain synthesis in lytically infected cells. MHC class I and MHC class II downregulation was mimicked by pharmacological inhibition of protein synthesis in latently infected cells. Our data suggest that although several mechanisms may contribute to MHC class I downregulation in the course of EBV replication, inhibition of MHC class I synthesis plays the primary role in the process. PMID- 17108040 TI - Immunogenicity and protective efficacy of DNA vaccines expressing rhesus cytomegalovirus glycoprotein B, phosphoprotein 65-2, and viral interleukin-10 in rhesus macaques. AB - Rhesus cytomegalovirus (RhCMV) infection of macaques exhibits strong similarities to human CMV (HCMV) persistence and pathogenesis. The immunogenicity of DNA vaccines encoding three RhCMV proteins (a truncated version of glycoprotein B lacking the transmembrane region and endodomain [gBDeltaTM], phosphoprotein 65-2 [pp65-2], and viral interleukin-10 [vIL-10]) was evaluated in rhesus macaques. Two groups of monkeys (four per group) were genetically immunized four times with a mixture of either pp65-2 and gBDeltaTM or pp65-2, vIL-10, and gBDeltaTM. The vaccinees developed anti-gB and anti-pp65-2 antibodies in addition to pp65-2 cellular responses after the second booster immunization, with rapid responses observed with subsequent DNA injections. Weak vIL-10 immune responses were detected in two of the four immunized animals. Neutralizing antibodies were detected in seven monkeys, although titers were weak compared to those observed in naturally infected animals. The immunized monkeys and naive controls were challenged intravenously with 10(5) PFU of RhCMV. Anamnestic binding and neutralizing antibody responses were observed 1 week postchallenge in the vaccinees. DNA vaccination-induced immune responses significantly decreased peak viral loads in the immunized animals compared to those in the controls. No difference in peak viral loads was observed between the pp65-2/gBDeltaTM DNA- and pp65-2/vIL-10/gBDeltaTM-vaccinated groups. Antibody responses to nonvaccine antigens were lower postchallenge in both vaccine groups than in the controls, suggesting long-term control of RhCMV protein expression. These data demonstrated that DNA vaccines targeting the RhCMV homologues of HCMV gB and pp65 altered the course of acute and persistent RhCMV infection in a primate host. PMID- 17108041 TI - Selective restriction of Nef-defective human immunodeficiency virus type 1 by a proteasome-dependent mechanism. AB - The Nef protein enhances human immunodeficiency virus type 1 (HIV-1) infectivity by facilitating an early postentry step in the virus life cycle. We report here that the addition of MG132 or lactacystin, each a specific inhibitor of cellular proteasome activity, preferentially enhances cellular permissiveness to infection by Nef-defective versus wild-type HIV-1. Pseudotyping by the glycoprotein of vesicular stomatitis virus rendered Nef-defective HIV-1 particles minimally responsive to the enhancing effects of proteasome inhibitors. These results suggest that Nef enhances the infectivity of HIV-1 particles by reducing their susceptibility to proteasomal degradation in target cells. PMID- 17108042 TI - Fc gamma receptors play a dominant role in protective tumor immunity against a virus-encoded tumor-specific antigen in a murine model of experimental pulmonary metastases. AB - Simian virus 40 (SV40) large tumor antigen (Tag) represents a virus-encoded tumor specific antigen expressed in many types of human cancers and a potential immunologic target for antitumor responses. Fc receptors are important mediators in the regulation and execution of host effector mechanisms against conditions including infectious diseases, autoimmunity, and cancer. By examining tumor protection in SV40 Tag-immunized wild-type BALB/c mice using an experimental pulmonary metastasis model, we attempted to address whether engagement of the immunoglobulin G Fc receptors (FcgammaRs) on effector cells is necessary to mediate antitumor responses. All immunized BALB/c FcgammaR-/- knockout mice developed anti-SV40 Tag antibody responses prior to experimental challenge with a tumorigenic cell line expressing SV40 Tag. However, all mice deficient in the activating FcgammaRI (CD64) and FcgammaRIII (CD16) were unable to mount protective immunologic responses against tumor challenge and developed tumor lung foci. In contrast, mice lacking the inhibitory receptor FcgammaRII (CD32) demonstrated resistance to tumorigenesis. These results underscore the importance of effector cell populations expressing FcgammaRI/III within this murine tumor model system, and along with the production of a specific humoral immune response, antibody-dependent cell-mediated cytotoxicity (ADCC) may be a functioning mechanism of tumor clearance. Additionally, these data demonstrate the potential utility of ADCC as a viable approach for targeting vaccination strategies that promote FcgammaRI/III scavenging pathways against cancer. PMID- 17108043 TI - CXCR3-dependent recruitment of antigen-specific T lymphocytes to the liver during murine cytomegalovirus infection. AB - Innate inflammatory events promoting antiviral defense in the liver against murine cytomegalovirus (MCMV) infection have been characterized. However, the mechanisms that regulate the selective recruitment of inflammatory T lymphocytes to the liver during MCMV infection have not been defined. The studies presented here demonstrate the expression of monokine induced by gamma interferon (IFN gamma; Mig/CXCL9) and IFN-gamma-inducible protein 10 (IP-10/CXCL10) in liver leukocytes and correlate their production with the infiltration of MCMV-specific CD8 T cells into the liver. Antibody-mediated neutralization of CXCL9 and CXCL10 and studies using mice deficient in CXCR3, the primary known receptor for these chemokines, revealed that CXCR3-dependent mechanisms promote the infiltration of virus-specific CD8 T cells into the liver during acute infection with MCMV. Furthermore, CXCR3 functions augmented the hepatic accumulation of CD8 T-cell IFN gamma responses to MCMV. Evaluation of protective functions demonstrated enhanced pathology that overlapped with transient increases in virus titers in CXCR3 deficient mice. However, ultimate viral clearance and survival were not compromised. Thus, CXCR3-mediated signals support the accumulation of MCMV specific CD8 T cells that contribute to, but are not exclusively required for, protective responses in a virus-infected tissue site. PMID- 17108044 TI - Tumor necrosis factor (TNF) protects resistant C57BL/6 mice against herpes simplex virus-induced encephalitis independently of signaling via TNF receptor 1 or 2. AB - Tumor necrosis factor (TNF) is a multifunctional cytokine that has a role in induction and regulation of host innate and adaptive immune responses. The importance of TNF antiviral mechanisms is reflected by the diverse strategies adopted by different viruses, particularly members of the herpesvirus family, to block TNF responses. TNF binds and signals through two receptors, Tnfrsf1a (TNF receptor 1 [TNFR1], or p55) and Tnfrsf1b (TNFR2, or p75). We report here that herpes simplex virus 1 (HSV-1) infection of TNF-/- mice on the resistant C57BL/6 genetic background results in significantly increased susceptibility (P < 0.0001, log rank test) to fatal HSV encephalitis (HSE) and prolonged persistence of elevated levels of virus in neural tissues. In contrast, although virus titers in neural tissues of p55-/- N13 mice were elevated to levels comparable to what was found for the TNF-/- mice, the p55-/- N13 mice were as resistant as control C57BL/6 mice (P > 0.05). The incidence of fatal HSE was significantly increased by in vivo neutralization of TNF using soluble TNFR1 (sTNFR1) or depletion of macrophages in C57BL/6 mice (P = 0.0038 and P = 0.0071, respectively). Strikingly, in vivo neutralization of TNF in HSV-1-infected p55-/- p75-/- mice by use of three independent approaches (treatment with soluble p55 receptor, anti TNF monoclonal antibody, or in vivo small interfering RNA against TNF) resulted in significantly increased mortality rates (P = 0.005), comparable in magnitude to those for C57BL/6 mice treated with sTNFR1 (P = 0.0018). Overall, these results indicate that while TNF is required for resistance to fatal HSE, both p55 and p75 receptors are dispensable. Precisely how TNF mediates protection against HSV-1 mortality in p55-/- p75-/- mice remains to be determined. PMID- 17108045 TI - Severe acute respiratory syndrome coronavirus protein 6 accelerates murine coronavirus infections. AB - One or more of the unique 3'-proximal open reading frames (ORFs) of the severe acute respiratory syndrome (SARS) coronavirus may encode determinants of virus virulence. A prime candidate is ORF6, which encodes a 63-amino-acid membrane associated peptide that can dramatically increase the lethality of an otherwise attenuated JHM strain of murine coronavirus (L. Pewe, H. Zhou, J. Netland, C. Tangudu, H. Olivares, L. Shi, D. Look, T. Gallagher, and S. Perlman, J. Virol. 79:11335-11342, 2005). To discern virulence mechanisms, we compared the in vitro growth properties of rJ.6, a recombinant JHM expressing the SARS peptide, with isogenic rJ.6-KO, which has an inactive ORF containing a mutated initiation codon and a termination codon at internal position 27. The rJ.6 infections proceeded rapidly, secreting progeny about 1.5 h earlier than rJ.6-KO infections did. The rJ.6 infections were also set apart by early viral protein accumulation and by robust expansion via syncytia, a characteristic feature of JHM virus dissemination. We found no evidence for protein 6 operating at the virus entry or assembly stage, as virions from either infection were indistinguishable. Rather, protein 6 appeared to operate by fostering viral RNA and protein synthesis, as RNA quantifications by reverse transcription-quantitative PCR revealed viral RNA levels in the rJ.6 cultures that were five to eight times higher than those lacking protein 6. Furthermore, protein 6 coimmunoprecipitated with viral RNAs and colocalized on cytoplasmic vesicles with replicating viral RNAs. The SARS coronavirus encodes a novel membrane protein 6 that can accelerate replication of a related mouse virus, a property that may explain its ability to increase in vivo virus virulence. PMID- 17108046 TI - Incorporation of glycosylphosphatidylinositol-anchored granulocyte- macrophage colony-stimulating factor or CD40 ligand enhances immunogenicity of chimeric simian immunodeficiency virus-like particles. AB - The rapid worldwide spread of human immunodeficiency virus (HIV) mandates the development of successful vaccination strategies. Since live attenuated HIV is not accepted as a vaccine due to safety concerns, virus-like particles (VLPs) offer an attractive safe alternative because they lack the viral genome yet they are perceived by the immune system as a virus particle. We hypothesized that adding immunostimulatory signals to VLPs would enhance their efficacy. To accomplish this we generated chimeric simian immunodeficiency virus (SIV) VLPs containing either glycosylphosphatidylinositol (GPI)-anchored granulocyte macrophage colony-stimulating factor (GM-CSF) or CD40 ligand (CD40L) and investigated their biological activity and ability to enhance immune responses in vivo. Immunization of mice with chimeric SIV VLPs containing GM-CSF induced SIV Env-specific antibodies as well as neutralizing activity at significantly higher levels than those induced by standard SIV VLPs, SIV VLPs containing CD40L, or standard VLPs mixed with soluble GM-CSF. In addition, mice immunized with chimeric SIV VLPs containing either GM-CSF or CD40L showed significantly increased CD4(+)- and CD8(+)-T-cell responses to SIV Env, compared to standard SIV VLPs. Taken together, these results demonstrate that the incorporation of immunostimulatory molecules enhances humoral and cellular immune responses. We propose that anchoring immunostimulatory molecules into SIV VLPs can be a promising approach to augmenting the efficacy of VLP antigens. PMID- 17108047 TI - Preferential activation of Toll-like receptor nine by CD46-utilizing adenoviruses. AB - Adenoviruses (Ads) are responsible for respiratory, ocular, and gastrointestinal illnesses in humans. While the majority of serotypes utilize coxsackievirus adenovirus receptor (CAR) as their primary attachment receptor, subgroup B and subgroup D Ad37 serotypes use CD46. Given the propensity of Ad vectors to activate host immune responses, we sought to investigate their potential for type I interferon induction. We found that CD46 Ads were capable of alpha interferon (IFN-alpha) induction by peripheral blood mononuclear cells and that plasmacytoid dendritic cells (pDCs) were the principal producers of this cytokine. IFN-alpha induction correlated with the permissivity of pDCs to CD46- but not CAR-utilizing Ad serotypes. A role for Toll-like receptor 9 (TLR9) recognition of Ad was supported by the requirement for viral DNA and efficient endosomal acidification and by the ability of a TLR9-inhibitory oligonucleotide to attenuate IFN-alpha induction. Cell lines expressing TLR9 that are permissive to infection by both CAR- and CD46-utilizing serotypes showed a preferential induction of TLR9 mediated events by CD46-utilizing Ads. Specifically, the latter virus types induced higher levels of cytokine expression and NF-kappaB activation in HeLa cells than CAR-dependent Ad types, despite equivalent infection rates. Therefore, infectivity alone is not sufficient for TLR9 activation, but this activation instead is regulated by a specific receptor entry pathway. These data reveal a novel mode of host immune recognition of Ad with implications for Ad pathogenesis and for the use of unconventional Ad vectors for gene delivery and vaccine development. PMID- 17108048 TI - Identification of a ribonucleoprotein intermediate of tomato mosaic virus RNA replication complex formation. AB - The replication of eukaryotic positive-strand RNA virus genomes occurs in the membrane-bound RNA replication complexes. Previously, we found that the extract of evacuolated tobacco BY-2 protoplasts (BYL) is capable of supporting the translation and subsequent replication of the genomic RNAs of plant positive strand RNA viruses, including Tomato mosaic virus (ToMV). Here, to dissect the process that precedes the formation of ToMV RNA replication complexes, we prepared membrane-depleted BYL (mdBYL), in which the membranes were removed by centrifugation. In mdBYL, ToMV RNA was translated to produce the 130-kDa and 180 kDa replication proteins, but the synthesis of any ToMV-related RNAs did not occur. When BYL membranes were added back to the ToMV RNA-translated mdBYL after the termination of translation with puromycin, ToMV RNA was replicated. Using a replication-competent ToMV derivative that encodes the FLAG-tagged 180-kDa replication protein, it was shown by affinity purification that a complex that contained the 130-kDa and 180-kDa proteins and ToMV genomic RNA was formed after translation in mdBYL. When the complex was mixed with BYL membranes, ToMV RNA was replicated, which suggests that this ribonucleoprotein complex is an intermediate of ToMV RNA replication complex formation. We have named this ribonucleoprotein complex the "pre-membrane-targeting complex." Our data suggest that the formation of the pre-membrane-targeting complex is coupled with the translation of ToMV RNA, while posttranslationally added exogenous 180-kDa protein and replication templates can contribute to replication and can be replicated, respectively. Based on these results, we discuss the mechanisms of ToMV RNA replication complex formation. PMID- 17108049 TI - Characterization of the uracil-DNA glycosylase activity of Epstein-Barr virus BKRF3 and its role in lytic viral DNA replication. AB - Uracil-DNA glycosylases (UDGs) of the uracil-N-glycosylase (UNG) family are the primary DNA repair enzymes responsible for removal of inappropriate uracil from DNA. Recent studies further suggest that the nuclear human UNG2 and the UDGs of large DNA viruses may coordinate with their DNA polymerase accessory factors to enhance DNA replication. Based on its amino acid sequence, the putative UDG of Epstein-Barr virus (EBV), BKRF3, belongs to the UNG family of proteins, and it was demonstrated previously to enhance oriLyt-dependent DNA replication in a cotransfection replication assay. However, the expression and enzyme activity of EBV BKRF3 have not yet been characterized. In this study, His-BKRF3 was expressed in bacteria and purified for biochemical analysis. Similar to the case for the Escherichia coli and human UNG enzymes, His-BKRF3 excised uracil from single stranded DNA more efficiently than from double-stranded DNA and was inhibited by the purified bacteriophage PBS1 inhibitor Ugi. In addition, BKRF3 was able to complement an E. coli ung mutant in rifampin and nalidixic acid resistance mutator assays. The expression kinetics and subcellular localization of BKRF3 products were detected in EBV-positive lymphoid and epithelial cells by using BKRF3-specific mouse antibodies. Expression of BKRF3 is regulated mainly by the immediate-early transcription activator Rta. The efficiency of EBV lytic DNA replication was slightly affected by BKRF3 small interfering RNA (siRNA), whereas cellular UNG2 siRNA or inhibition of cellular and viral UNG activities by expressing Ugi repressed EBV lytic DNA replication. Taking these results together, we demonstrate the UNG activity of BKRF3 in vitro and in vivo and suggest that UNGs may participate in DNA replication or repair and thereby promote efficient production of viral DNA. PMID- 17108050 TI - GLUT1 is not the primary binding receptor but is associated with cell-to-cell transmission of human T-cell leukemia virus type 1. AB - GLUT1 has recently been suggested to be a binding receptor for human T-cell leukemia virus type 1 (HTLV-1). We used a novel, short-term assay to define the role of GLUT1 in cell-to-cell transmission. Although increasing cell surface levels of GLUT1 enhanced HTLV-I transfer, efficient virus spread correlated largely with heparan sulfate proteoglycan (HSPG) expression on target cells. Moreover, since activated CD4+ T cells and cord blood lymphocytes that are susceptible to HTLV-1 infection expressed undetectable levels of surface GLUT1, these results indicate that GLUT1 and HSPGs are important for efficient cell-to cell transmission of HTLV-1 but raise concerns on the role of GLUT1 as the HTLV-1 primary binding receptor. PMID- 17108051 TI - HLA class I binding promiscuity of the CD8 T-cell epitopes of human papillomavirus type 16 E6 protein. AB - One of the critical steps in the progression to cervical cancer appears to be the establishment of persistent human papillomavirus (HPV) infection. We have demonstrated that the lack of cytotoxic T-lymphocyte response to HPV type 16 (HPV 16) E6 protein was associated with persistence and that the potential presence of dominant CD8 T-cell epitopes was most frequently found (n = 4 of 23) in the E6 16 40 region by examining the pattern of CD8 T-cell epitopes within the E6 protein in women who had cleared their HPV 16 infections. The goal of this study was to define the minimal/optimal amino acid sequences and the HLA restricting molecules of these dominant CD8 T-cell epitopes as well as those of subdominant ones if present. Three dominant epitopes, E6 29-38 (TIHDIILECV; restricted by the HLA A0201 molecule), E6 29-37 (TIHDIILEC; restricted by B48), and E6 31-38 (HDIILECV; restricted by B4002), and one subdominant epitope, E6 52-61 (FAFRDLCIVY; restricted by B35) were characterized. Taken together with a previously described dominant epitope, E6 52-61 (FAFRDLCIVY; restricted by B57), the CD8 T-cell epitopes demonstrated striking HLA class I binding promiscuity. All of these epitopes were endogenously processed, but the presence of only two of the five epitopes could have been predicted based on the known binding motifs. The HLA class I promiscuity which has been described for human immunodeficiency virus may be more common than previously recognized. PMID- 17108052 TI - Human immunodeficiency virus type 1 matrix protein assembles on membranes as a hexamer. AB - The membrane-binding matrix (MA) domain of the human immunodeficiency virus type 1 (HIV-1) structural precursor Gag (PrGag) protein oligomerizes in solution as a trimer and crystallizes in three dimensions as a trimer unit. A number of models have been proposed to explain how MA trimers might align with respect to PrGag capsid (CA) N-terminal domains (NTDs), which assemble hexagonal lattices. We have examined the binding of naturally myristoylated HIV-1 matrix (MyrMA) and matrix plus capsid (MyrMACA) proteins on membranes in vitro. Unexpectedly, MyrMA and MyrMACA proteins both assembled hexagonal cage lattices on phosphatidylserine cholesterol membranes. Membrane-bound MyrMA proteins did not organize into trimer units but, rather, organized into hexamer rings. Our results yield a model in which MA domains stack directly above NTD hexamers in immature particles, and they have implications for HIV assembly and interactions between MA and the viral membrane glycoproteins. PMID- 17108053 TI - Kaposi's sarcoma-associated herpesvirus-encoded protein kinase and its interaction with K-bZIP. AB - The oncogenic herpesvirus, Kaposi's sarcoma-associated herpesvirus, also identified as human herpesvirus 8, contains genes producing proteins that control transcription and influence cell signaling. Open reading frame 36 (ORF36) of this virus encodes a serine/threonine protein kinase, which is designated the viral protein kinase (vPK). Our recent efforts to elucidate the role of vPK in the viral life cycle have focused on identifying viral protein substrates and determining the effects of vPK-mediated phosphorylation on specific steps in viral replication. The vPK gene was transcribed into 4.2-kb and 3.6-kb mRNAs during the early and late phases of viral reactivation. vPK is colocalized with viral DNA replication/transcription compartments as marked by a polymerase processivity factor, and K-bZIP, a protein known to bind the viral DNA replication origin (Ori-Lyt) and to regulate viral transcription. The vPK physically associated with and strongly phosphorylated K-bZIP at threonine 111, a site also recognized by the cyclin-dependent kinase Cdk2. Both K-bZIP and vPK were corecruited to viral promoters targeted by K-bZIP as well as to the Ori-Lyt region. Phosphorylation of K-bZIP by vPK had a negative impact on K-bZIP transcription repression activity. The extent of posttranslational modification of K-bZIP by sumoylation, a process that influences its repression function, was decreased by vPK phosphorylation at threonine 111. Our data thus identify a new role of vPK as a modulator of viral transcription. PMID- 17108054 TI - Nicotine and serotonin in immune regulation and inflammatory processes: a perspective. AB - Nicotine and serotonin modulate the innate and adaptive immune responses and the inflammatory states. Several nicotinic cholinergic and serotonergic receptor subtypes have been characterized in B and T lymphocytes, monocytes, macrophages, and dendritic cells. The use of knockout mice has allowed a better characterization of nicotinic receptors and their role in anti-inflammatory processes in these cells. Cytokines play a crucial role in controlling inflammatory reactions. Nicotine and serotonin have been reported to regulate cytokine release. Cholinergic mechanisms also play an important role in inflammation through endogenous acetylcholine. Nicotine mimics this effect by activating the cholinergic anti-inflammatory pathways. New concepts of reciprocal interactions between nicotine and serotonin are emerging. The role of nicotine as an anti-inflammatory agent has been established, whereas that of serotonin remains more controversial. PMID- 17108056 TI - The human EGF-TM7 receptor EMR3 is a marker for mature granulocytes. AB - EMR3 is a member of the epidermal growth factor-seven-transmembrane (EGF-TM7) family of adhesion class TM7 receptors. This family also comprises CD97, EMR1, EMR2, and EMR4. To characterize human EMR3 at the protein level, we generated Armenian hamster mAb. Using the mAb 3D7, we here demonstrate that EMR3, like other EGF-TM7 receptors, is expressed at the cell surface as a heterodimeric molecule consisting of a long extracellular alpha-chain, which possesses at its N terminus EGF-like domains and a membrane-spanning beta-chain. Flow cytometric analysis revealed that all types of myeloid cells express EMR3. In peripheral blood, the highest expression of EMR3 was found on granulocytes. More mature CD16(+) monocytes express high levels of EMR3, and CD16(-) monocytes and myeloid dendritic cells (DC) are EMR3(dim/low). Lymphocytes and plasmacytoid DC are EMR3( ). It is interesting that in contrast with CD97 and EMR2, CD34(+)CD33(-)/CD38(-) committed hematopoietic stem cells and CD34(+)CD33(+)/CD38(+) progenitors in bone marrow do not express EMR3. In vitro differentiation of HL-60 cells and CD34(+) progenitor cells revealed that EMR3 is only up-regulated during late granulopoiesis. These results demonstrate that the expression of EGF-TM7 receptors on myeloid cells is differentially regulated. EMR3 is the first family member found mainly on granulocytes. PMID- 17108057 TI - Triethylenetetramine and metabolites: levels in relation to copper and zinc excretion in urine of healthy volunteers and type 2 diabetic patients. AB - Triethylenetetramine (TETA), a selective Cu(II)-chelator used in the treatment of Wilson's disease, is now undergoing clinical trials for the treatment of heart failure in diabetes. Despite decades of clinical use, knowledge of its pharmacology in human subjects remains incomplete. Here, we first used liquid chromatography-mass spectrometry (LC-MS) to detect and identify major metabolites of TETA in human plasma and urine, and then used this method to measure concentrations of TETA and its metabolites in the urine of healthy and diabetic subjects who were administered increasing doses (300, 600, 1200, and 2400 mg) of TETA orally. Twenty-four-hour urine collections were performed before and after dosing participants. Two major metabolites of TETA were detected in human urine, N(1)-acetyltriethylenetetramine (MAT) and N(1),N(10) diacetyltriethylenetetramine, the latter being novel. Both metabolites were verified with synthetic standards by LC-MS. The proportion of unchanged TETA excreted as a fraction of total urinary drug-derived molecules was significantly higher in healthy than in matched diabetic subjects, consistent with a higher rate of TETA metabolism in the latter. TETA-evoked increases in urinary Cu excretion in nondiabetic subjects were more closely correlated with parent drug concentrations than in diabetic subjects, whereas, by contrast, urinary Cu was more closely associated with the sum of TETA and MAT. These findings are consistent with the hypothesis that MAT could play a significant role in the molecular mechanism by which TETA extracts Cu(II) from the systemic compartment in diabetic subjects. PMID- 17108058 TI - Induction of cytochrome P450: assessment in an immortalized human hepatocyte cell line (Fa2N4) using a novel higher throughput cocktail assay. AB - Over recent years the application of cocktail studies to measure biological markers has become increasingly popular. The current study investigated a novel approach in assessing cytochrome P450 (P450) enzyme induction in an immortalized cell line using a cocktail of five P450 substrate probes compared with the traditional single-probe approach. The findings reported herein support use of a cocktail approach to assess the induction of the major P450s, namely, CYP3A4, CYP1A2, and CYP2C9. CYP2C19 and CYP2D6 could also be followed as part of the cocktail approach reported. Response to prototypical inducers did not differ to those observed in the presence of the specific probes alone. Consequently, this approach requires significantly fewer sample numbers if screening the induction potential of more than one P450. Moreover, these studies highlight the utility of the immortalized cell line Fa2N4 as a robust model system for induction studies. In conclusion, the current experimental setup is an improvement on current approaches used to assess P450 induction, significantly increasing sample throughput. PMID- 17108059 TI - Flavonoids modulate monocarboxylate transporter-1-mediated transport of gamma hydroxybutyrate in vitro and in vivo. AB - The objective of this study was to determine the effects of flavonoids on the in vitro monocarboxylate transporter 1 (MCT1)-mediated transport and in vivo disposition of the drug of abuse, gamma-hydroxybutyrate (GHB). The uptake of GHB in rat MCT1 gene-transfected MDA-MB231 cells was significantly decreased in the presence of the flavonoids apigenin, biochanin A, chrysin, diosemin, fisetin, genistein, hesperitin, kaempferol, luteolin, morin, narigenin, phloretin, and quercetin, but was not affected by the flavonoid glycosides phloridzin and rutin. The IC(50) values for luteolin, morin, and phloretin were 0.41 +/- 0.14, 6.41 +/- 2.01, and 2.57 +/- 0.48 microM, with the inhibition mechanism for luteolin being competitive. [(3)H]Kaempferol and [(3)H]biochanin A did not exhibit MCT1-mediated uptake, suggesting that these flavonoids are not substrates for MCT1. The combination of luteolin and phloretin inhibited the uptake of GHB in a synergistic manner; however, the combination of luteolin and morin was antagonistic. GHB 1000 mg/kg was administered to rats by i.v. bolus, with or without the concomitant administration of luteolin 10 mg/kg i.v. After luteolin treatment, the renal and total clearances of GHB were significantly increased, probably because of inhibition of the MCT1-mediated renal reabsorption of GHB, and the sleep time significantly decreased (121 +/- 5 min versus 165 +/- 10 min) compared with control rats. Overall, the results of this study indicate that flavonoids from food or herbal products may significantly alter the pharmacokinetics and pharmacodynamics of MCT substrates. PMID- 17108060 TI - Effects of green tea compounds on irinotecan metabolism. AB - The effects of green tea compounds on the metabolism of irinotecan have never been investigated. We aimed to study whether catechins [(-)-epigallocatechin gallate (EGCG), (-)-epicatechin gallate (ECG), (-)-epigallocatechin (EGC), (-) epicatechin] affect the inactivation metabolism of irinotecan into 7-ethyl-10-[4 N-(1-piperidino)-1-amino]carbonyloxycamptothecin (NPC) (by CYP3A4) and 7-ethyl-10 hydroxycamptothecin (SN-38) into 7-ethyl-10-hydroxycamptothecin glucuronide (SN 38G) (by UGT1A1). Human liver microsomes, hepatocytes and Hep G2 cells were incubated with catechins and treated with irinotecan and/or SN-38. NPC and SN-38G formation was measured by high-performance liquid chromatography. UGT1A1 mRNA levels were measured by real-time polymerase chain reaction. In human liver microsomes, a concentration-dependent decrease in the formation of NPC and SN-38G was observed. In human hepatocytes, a significant increase in SN-38G production was observed in 33% (EGCG), 44% (ECG), and 44% (EGC) of the hepatocyte preparations. Phenobarbital increased the formation of SN-38G in 100% of the same hepatocyte preparations. In Hep G2 cells, no increase in SN-38G formation was observed. With the exception of ECG in one liver, catechins did not increase UGT1A1 mRNA levels. NPC production was also significantly increased in 40% of the hepatocyte preparations for each catechin. However, the production of 6beta hydroxytestosterone remained unaffected in other hepatocyte preparations. At pharmacologically relevant concentrations, catechins are unlikely to inhibit the formation of irinotecan inactive metabolites when administered concomitantly. The induction effect of catechins on UGT1A1 seems to be modest and highly variable. Catechins do not induce CYP3A4 activity. The effect of acute and prolonged use of green tea on the pharmacokinetics of irinotecan in patients remains to be evaluated. PMID- 17108061 TI - Effects of alterations in CAR on bilirubin detoxification in mouse collagen induced arthritis. AB - Nuclear receptors such as constitutive androstane receptor (CAR) and pregnane X receptor (PXR) regulate the transcription of cytochromes P450 and transporters. We investigated whether quantitative and functional changes in CAR and PXR could affect bilirubin detoxification in chronic arthritis. The CAR mRNA level was significantly decreased in the liver of mice with collagen-induced arthritis (CIA) compared with control mice. In normal mice treated with CAR agonists, relatively rapid elimination of bilirubin was observed after its intravenous injection. Next, we investigated the effects of CAR on bilirubin-detoxifying enzymes and transporters in arthritis. The mRNA levels of organic anion transporter peptide (OATP) 2, glutathione S-transferase (GST) A1, and GSTA2 were decreased in CIA mice, whereas the mRNA levels of OATP4, UDP-glucuronosyl transferase 1A1, and multidrug resistance-associated protein 2 remained unchanged. The protein levels and transport activities of OATP2 were also decreased in CIA mice. Furthermore, the CIA mice actually exhibited retarded elimination of bilirubin after its intravenous injection. These results indicate that alterations to CAR during arthritis affect the elimination of bilirubin because of changes in multiple bilirubin-detoxifying enzymes and transporters. PMID- 17108062 TI - The impact of thrombocytopenia from temozolomide and radiation in newly diagnosed adults with high-grade gliomas. AB - Temozolomide (TMZ) administered daily with radiation therapy (RT) for six weeks, followed by adjuvant TMZ for six months, has become standard therapy for patients with glioblastoma multiforme (GBM). After several newly diagnosed patients at our institution developed severe (grade 3-4), prolonged thrombocytopenia, we conducted a retrospective review to define the incidence, depth, and duration of thrombocytopenia associated with this therapy. We reviewed the medical records and laboratory data of all adult patients with newly diagnosed high-grade gliomas who started treatment with this regimen between June 2004, when the regimen was first used at our institution, and August 2005. Of the 52 patients who met the criteria for this review, grade 3-4 thrombocytopenia occurred in 10 (19%; 95% CI, 10%-33%). In eight patients, the thrombocytopenia was attributable to concurrent daily TMZ and RT. The median duration of grade 3-4 thrombocytopenia was 32 days (range, 1-389 days). Five patients (10%) required platelet transfusions, two (4%) have required continued biweekly platelet transfusions for over six months, and nine (17%) discontinued therapy because of thrombocytopenia. Grade 3-4 thrombocytopenia occurred in 25% of women and 14% of men. Grade 3-4 neutropenia and anemia were noted in 10% and 8% of patients, respectively, and were not clinically significant. Between 15% and 20% of our newly diagnosed patients receiving TMZ and RT developed severe (grade 3-4) and potentially irreversible thrombocytopenia. The factors that predispose patients to this toxicity have yet to be determined. This toxicity should be considered when (1) prescribing this regimen to patient populations where a clinical benefit has yet to be shown, (2) contemplating empirical escalations of the dose or duration of TMZ, or (3) combining it with other potentially myelosuppressive therapies. PMID- 17108063 TI - The relationship between six-month progression-free survival and 12-month overall survival end points for phase II trials in patients with glioblastoma multiforme. AB - Common end points for phase II trials in patients with glioblastoma multiforme (GBM) are six-month progression-free survival (PFS6) and 12-month overall survival (OS12). OS12 can be accurately measured but may be confounded with subsequent therapies upon progression, whereas the converse is true for PFS6. Our goal was to assess the relationship between these end points separately for phase II trials in patients with newly diagnosed GBM and patients with recurrent GBM. Data were pooled from 11 North Central Cancer Treatment Group trials for patients with newly diagnosed GBM (n = 1348). All patients received radiotherapy and pharmaceutical therapy (before, during, or after radiotherapy). Data were pooled from 16 trials that used various pharmaceuticals in treating patients for recurrent GBM (n = 345). All trial regimens were declared nonefficacious by predefined criteria. Overall per-patient concordance was estimated with a kappa statistic. The relationship between OS12 and PFS6 across study arms was assessed by weighted linear regression and Pearson's correlation. Simulation was used to determine the agreement of study outcomes when using PFS6 versus OS12 end points. Cox models with progression status as a time-dependent variable and Kaplan-Meier estimators were used to ascertain the association between progression-free survival status and overall survival. At present, 97% of the patients with newly diagnosed GBM and 95% of those with recurrent GBM have died. The PFS6 and OS12 were 43% and 41%, respectively, for patients with newly diagnosed disease and 9% and 14% for patients with recurrent disease. There was only moderate concordance between the end points on both the patient level and the study level. For the simulation studies, we established phase II efficacy criteria for each end point by using the pooled estimates of OS12 (PFS6) as historical controls. The study decisions made using PFS6 and OS12 were in agreement 88% and 90% of the time for the trials of newly diagnosed and recurrent disease, respectively. Finally, there was a strong association between progression-free survival status and overall survival. PFS6 seems to be a reasonable end point for phase II trials in patients with recurrent glioblastoma. PMID- 17108064 TI - CXCR4 and SDF1 expression in human meningiomas: a proliferative role in tumoral meningothelial cells in vitro. AB - Chemokines participate in cellular processes associated with tumor proliferation, migration, and angiogenesis. We previously demonstrated that stromal cell-derived factor 1 (SDF1) exerts a mitogenic activity in glioblastomas through the activation of its receptor CXCR4. Here we studied the expression of this chemokine in human meningiomas and its possible role in cell proliferation. Reverse transcriptase-PCR analysis for CXCR4 and SDF1 was performed on 55 human meningiomas (47 WHO grade I, 5 WHO II, and 3 WHO III). Immunolabeling for CXCR4 and SDF1 was performed on paraffin-embedded sections of these tumors. [(3)H]Thymidine uptake and Western blot analyses were performed on primary meningeal cell cultures of tumors to evaluate the proliferative activity of human SDF1alpha (hSDF1alpha) in vitro and the involvement of extracellular signal regulated kinase 1/2 (ERK1/2) activation in this process. CXCR4 mRNA was expressed by 78% of the tumor specimens and SDF1 mRNA by 53%. CXCR4 and SDF1 were often detected in the same tumor tissues and colocalized with epithelial membrane antigen immunostaining. In 9 of 12 primary cultures from meningiomas, hSDF1alpha induced significant cell proliferation that was strongly reduced by the mitogen activated protein kinase kinase inhibitor PD98059, involving ERK1/2 activation in the proliferative signal of hSDF1alpha. In fact, CXCR4 stimulation led to ERK1/2 phosphorylation/activation. In addition, the hSDF1alpha-induced cell proliferation was significantly correlated with the MIB1 staining index in the corresponding surgical specimen. In conclusion, we found that human meningiomas express CXCR4 and SDF1 and that hSDF1alpha induces proliferation in primary meningioma cell cultures through the activation of ERK1/2. PMID- 17108065 TI - Phase II study of Cloretazine for the treatment of adults with recurrent glioblastoma multiforme. AB - Cloretazine (VNP40101M) is a newly synthesized alkylating agent belonging to a novel class of alkylating agents called 1,2-bis(sulfonyl)hydrazines. Agents that belong to this class do not produce vinylating and chloroethylating species, and hence this class of alkylating agents is thought to have minimal systemic toxicity. Cloretazine produces two short-lived active species: 1,2 bis(methylsulfonyl)-1-(2-chloroethyl) hydrazine (a chloroethylating species) and a thiophilic carbamoylating methylisocyanate species. The chloroethylating species preferentially produces lesions at the O(6) position of guanine. The methylisocyanate species may inhibit O(6)-alkylguanine-DNA alkyltransferase, an important mechanism of resistance against alkylating agents. The purpose of this study was to determine the efficacy and tolerability of Cloretazine in patients with recurrent glioblastoma multiforme. The basis for the determination of efficacy was the proportion of patients alive without evidence of disease progression six months after initiation of treatment. Patients with recurrent glioblastoma multiforme received Cloretazine (300 mg/m(2)) intravenously every six weeks. Radiographic response, survival data, and toxicity were assessed. Thirty-two patients were enrolled. Median age was 56 years; 24 patients (75%) were men. At six months, two patients were alive and progression free, so the six month progression-free survival (PFS) was 6%. The median PFS was 6.3 weeks. There were no objective radiographic responses. Twelve patients had stable disease for at least one cycle, but only two patients received more than three cycles. Nine patients experienced grade 4 thrombocytopenia and three patients experienced grade 4 neutropenia. Cloretazine administered every six weeks was relatively well tolerated, although this schedule has insignificant activity for patients with recurrent glioblastoma multiforme. PMID- 17108066 TI - Successful treatment of childhood intramedullary spinal cord astrocytomas with irinotecan and cisplatin. AB - Childhood spinal cord astrocytomas are rare diseases, and their management is controversial. We report here our successful experience using irinotecan and cisplatin in three consecutive infants with progressing intramedullary astrocytomas. The first patient was a 16-month-old girl who presented with a grade III intramedullary astrocytoma that rapidly progressed after surgery and adjuvant chemotherapy. Weekly irinotecan (50 mg/m(2)) and cisplatin (30 mg/m(2)) for four consecutive weeks (one cycle) for a total of four cycles (I/C regimen) was used in order to avoid or delay radiotherapy. Radiological complete remission was achieved 10 months after completion of therapy, and 3.5 years after diagnosis the patient remains disease free. The second patient was a 19-month-old boy with a C3-T4 grade II intramedullary astrocytoma who received up-front vincristine and carboplatin for two months but remained clinically symptomatic. A followup MRI showed a larger tumor, and the patient was switched to the I/C regimen. A marked clinical improvement occurred after the first cycle, and MRI showed a very good partial remission at the end of therapy. At 16 months after diagnosis, the patient remains disease free. The third patient was a 10-month-old girl with a C2 T3 grade II intramedullary astrocytoma. She presented with severe pain that became steroid dependent during the month she was treated with the vincristine carboplatin regimen. When she was switched to the I/C regimen, the clinical symptoms responded within days. MRI at the end of therapy showed a significant reduction in tumor size, and one year after diagnosis the patient remains symptom free. Using this I/C regimen for childhood intramedullary astrocytoma, we obtained remarkable clinicoradiological responses while avoiding the use of radiotherapy. PMID- 17108067 TI - Epidemiologic, clinical, and diagnostic aspects of coccidioidomycosis. PMID- 17108068 TI - Horizontal gene transfer in a polyclonal outbreak of carbapenem-resistant Acinetobacter baumannii. AB - In the last few years, phenotypically carbapenem resistant Acinetobacter strains have been identified throughout the world, including in many of the hospitals and intensive care units (ICUs) of Australia. Genotyping of Australian ICU outbreak associated isolates by pulsed-field gel electrophoresis of whole genomic DNA indicated that different strains were cocirculating within one hospital. The carbapenem-resistant phenotype of these and other Australian isolates was found to be due to carbapenem-hydrolyzing activity associated with the presence of the blaOXA-23 gene. In all resistant strains examined, the blaOXA-23 gene was adjacent to the insertion sequence ISAba1 in a structure that has been found in Acinetobacter baumannii strains of a similar phenotype from around the world; blaOXA-51-like genes were also found in all A. baumannii strains but were not consistently associated with ISAba1, which is believed to provide the promoter required for expression of linked antibiotic resistance genes. Most isolates were also found to contain additional antibiotic resistance genes within the cassette arrays of class 1 integrons. The same cassette arrays, in addition to the ISAba1 blaOXA-23 structure, were found within unrelated strains, but no common plasmid carrying these accessory genetic elements could be identified. It therefore appears that antibiotic resistance genes are readily exchanged between cocirculating strains in epidemics of phenotypically indistinguishable organisms. Epidemiological investigation of major outbreaks should include whole-genome typing as well as analysis of potentially transmissible resistance genes and their vehicles. PMID- 17108069 TI - A multicenter study evaluating the current strategies for isolating Staphylococcus aureus strains with reduced susceptibility to glycopeptides. AB - Glycopeptide-intermediate Staphylococcus aureus (GISA) and heterogeneous GISA (hGISA) strains are notoriously difficult to detect in the diagnostic laboratory. The clinical importance of GISA, and particularly hGISA, will only be obvious when a definitive detection method is available. A few novel GISA and hGISA detection methods have been proposed; however, their validity has never been tested on a significant scale and in different laboratories. This study compares three screening methods for detecting GISA and hGISA strains in 12 laboratories, using a blind panel of 48 strains with known glycopeptide susceptibilities. The three screening methods used were brain heart infusion agar with 6 mg/liter vancomycin (BHIA6V) (CDC/CLSI), Mueller-Hinton agar with 5 mg/liter teicoplanin (MHA5T) (European Antimicrobial Resistance Surveillance System [EARSS]), and the macrodilution method Etest (MET) (EARSS), with population analysis profile-area under the curve analysis as the gold standard. Sensitivity and specificity were highest for MHA5T and MET, which identified 82.5% and 85.9% of strains, respectively. BHIA6V had poor sensitivity, particularly for hGISA (11.5% of strains were detected), and gave the largest interlaboratory variation in performance. MET exhibited the least interlaboratory variation. It is essential that laboratories use appropriate methods to detect GISA/hGISA strains so that the prevalence and clinical importance of these strains can be assessed properly. PMID- 17108070 TI - Veillonella parvula discitis and secondary bacteremia: a rare infection complicating endoscopy and colonoscopy? AB - We report a case of Veillonella parvula lumbar discitis and secondary bacteremia confirmed by molecular characterization of the 16S rRNA genes. Identification of the organism was essential for an appropriate choice of antimicrobial therapy following the failure of empirical flucloxacillin. Veillonella spp. are normal flora of the gastrointestinal tract, raising the possibility that an endoscopy and colonoscopy performed 8 weeks prior to presentation, during which small intestinal and rectal biopsies were obtained, was the portal of entry. This case highlights the importance of obtaining a microbiologic diagnosis, particularly in patients who previously have had procedures involving instrumentation. PMID- 17108071 TI - CTX-M beta-lactamase-producing Escherichia coli in French hospitals: prevalence, molecular epidemiology, and risk factors. AB - In 2004, 65 CTX-M-producing Escherichia coli isolates were collected from infected patients in four French hospitals. The blaCTX-M-15 genes were predominant. Pulsed-field gel electrophoresis highlighted a clonal propagation of CTX-M-15-producing strains belonging to phylogenetic group B2, notably in the community. The main risk factors for acquiring these isolates were urinary tract infections or the presence of a urinary catheter in diabetic or renal failure patients. PMID- 17108072 TI - Prevalence and clinical significance of Staphylococcus aureus small-colony variants in cystic fibrosis lung disease. AB - Small-colony variants (SCVs) of Staphylococcus aureus can be isolated from the chronically infected airways of patients suffering from cystic fibrosis (CF). These slow-growing morphological variants have been associated with persistent and antibiotic-resistant infections, such as osteomyelitis and device-related infections, but no information is available to date regarding the clinical significance of this special phenotype in CF lung disease. We therefore investigated the prevalence of S. aureus SCVs in CF lung disease in a 12-month prospective study and correlated the microbiological culture results with the patients' clinical data. A total of 252 patients were screened for the presence of SCVs. The prevalence rate was determined to be 17% (95% confidence interval, 10 to 25%) among S. aureus carriers. S. aureus isolates with the SCV phenotype showed significantly higher antibiotic resistance rates than those with the normal phenotype. Patients positive for SCVs were significantly older (P = 0.0099), more commonly cocolonized with Pseudomonas aeruginosa (P = 0.0454), and showed signs of more advanced disease, such as lower forced expiratory volume in 1 s (P = 0.0148) than patients harboring S. aureus with a solely normal phenotype. The logistic regression model determined lower weight (P = 0.016), advanced age (P = 0.000), and prior use of trimethoprim-sulfamethoxazole (P = 0.002) as independent risk factors for S. aureus SCV positivity. The clinical status of CF patients is known to be affected by multiple parameters. Nonetheless, the independent risk factors determined here point to the impact of S. aureus SCVs on chronic and persistent infections in advanced CF lung disease. PMID- 17108073 TI - Fast DNA serotyping of Escherichia coli by use of an oligonucleotide microarray. AB - Classical antibody-based serotyping of Escherichia coli is an important method in diagnostic microbiology for epidemiological purposes, as well as for a rough virulence assessment. However, serotyping is so tedious that its use is restricted to a few reference laboratories. To improve this situation we developed and validated a genetic approach for serotyping based on the microarray technology. The genes encoding the O-antigen flippase (wzx) and the O-antigen polymerase (wzy) were selected as target sequences for the O antigen, whereas fliC and related genes, which code for the flagellar monomer, were chosen as representatives for the H phenotype. Starting with a detailed bioinformatic analysis and oligonucleotide design, an ArrayTube-based assay was established: a fast and robust DNA extraction method was coupled with a site-specific, linear multiplex labeling procedure and hybridization analysis of the biotinylated amplicons. The microarray contained oligonucleotide DNA probes, each in duplicate, representing 24 of the epidemiologically most relevant of the over 180 known O antigens (O antigens 4, 6 to 9, 15, 26, 52, 53, 55, 79, 86, 91, 101, 103, 104, 111, 113, 114, 121, 128, 145, 157, and 172) as well as 47 of the 53 different H antigens (H antigens 1 to 12, 14 to 16, 18 to 21, 23 to 34, 37 to 43, 45, 46, 48, 49, 51 to 54, and 56). Evaluation of the microarray with a set of defined strains representing all O and H serotypes covered revealed that it has a high sensitivity and a high specificity. All of the conventionally typed 24 O groups and all of the 47 H serotypes were correctly identified. Moreover, strains which were nonmotile or nontypeable by previous serotyping assays yielded unequivocal results with the novel ArrayTube assay, which proved to be a valuable alternative to classical serotyping, allowing processing of single colonies within a single working day. PMID- 17108074 TI - Use of partial 16S rRNA gene sequencing for identification of Legionella pneumophila and non-pneumophila Legionella spp. AB - We examined 49 Legionella species, 26 L. pneumophila and 23 non-pneumophila Legionella spp., using partial 16S rRNA gene sequencing. This approach accurately identified all the L. pneumophila isolates, characterized all non-pneumophila Legionella isolates as such within this genus, and classified most (20/23; 87%) of the non-pneumophila Legionella isolates to the species level. PMID- 17108075 TI - Development of multiplex real-time reverse transcriptase PCR assays for detecting eight medically important flaviviruses in mosquitoes. AB - A multiplex real-time reverse transcriptase PCR has been developed for the rapid detection and identification of eight medically important flaviviruses from laboratory-reared, virus-infected mosquito pools. The method used involves the gene-specific amplification of yellow fever virus (YFV), Japanese encephalitis virus (JEV), West Nile virus (WNV), St. Louis encephalitis virus (SLEV), and dengue virus (DENV) serotypes 1 to 4 (DENV-1 to DENV-4, respectively) by use of the flavivirus consensus amplimers located at the RNA-dependent RNA polymerase domain of nonstructural protein 5. Virus-specific amplicons were detected by four newly characterized TaqMan fluorogenic probes (probes specific for YFV, JEV, WNV, and SLEV) and four previously published probes specific for DENV-1 to -4 (L. J. Chien, T. L. Liao, P. Y. Shu, J. H. Huang, D. J. Gubler, and G. J. Chang, J. Clin. Microbiol. 44:1295-1304, 2006). This assay had a specificity of 100% and various sensitivities of at least 3.5 PFU/ml for YFV, 2.0 PFU/ml for JEV, 10.0 PFU/ml for WNV, and 10.0 PFU/ml for SLEV. Additionally, we have developed an in vitro transcription system to generate RNase-resistant RNA templates for each of these eight viruses. These templates can be incorporated into the assay as RNA copy number controls and/or as external controls for RNA-spiked mosquito pools for quality assurance purposes. Although further study with mosquitoes collected in the field is needed, the incorporation of this assay into mosquito surveillance could be used as an early-warning system for the detection of medically important flaviviruses, particularly when the cocirculation of multiple viruses in the same region is suspected. PMID- 17108076 TI - Analysis of changes in recent tuberculosis transmission patterns after a sharp increase in immigration. AB - We conducted a population-based molecular epidemiological study of tuberculosis (TB) in Madrid, Spain (2002 to 2004), to define transmission patterns and factors associated with clustering. We particularly focused on examining how the increase in TB cases among immigrants in recent years (2.8% in 1997 to 1999 to 36.2% during the current study) was modifying transmission patterns. Mycobacterium tuberculosis isolates obtained from patients living in nine districts of Madrid (1,459,232 inhabitants) were genotyped. The TB case rate among foreign-born people was three to four times that of Spanish-born people, and the median time from arrival to the onset of treatment was 22.4 months. During the study period, 227 (36.3%) patients were grouped in 64 clusters, and 115 (50.7%) of them were in 21 clusters with mixed Spanish-born and foreign-born patients. Three of the 21 mixed clusters accounted for 21.1% of clustered patients. Twenty-two of 38 (57.9%) immigrants in mixed clusters were infected with TB strains that had already been identified in the native population in 1997 to 1999, including the three most prevalent strains. Factors identified as independent predictors of clustering were homelessness (odds ratio [OR], 2.3; 95% confidence interval [95% CI], 1.2 to 4.5; P = 0.011) and to be born in Spain (OR, 1.8; 95% CI, 1.2 to 2.6; P = 0.002). The results indicated that (i) TB transmission was higher in Spanish born people, associated mainly with homelessness, (ii) that foreign-born people were much less likely to be clustered, suggesting a higher percentage of infection before arriving in Spain, and (iii) that an extensive transmission between Spanish- and foreign-born populations, caused mainly by autochthonous strains, was taking place in Madrid. PMID- 17108077 TI - Detection of Coccidioides species in clinical specimens by real-time PCR. AB - Coccidioides spp. are dimorphic fungal pathogens endemic to the semiarid regions of North, Central, and South America. Currently, direct smear and culture are the most common means of identifying Coccidioides spp. While these methods offer relatively sensitive and specific means of detecting Coccidioides spp., growth in culture may take up to 3 weeks, potentially delaying the diagnosis and initiation of appropriate antifungal therapy. In addition, growth of the organism represents a significant safety risk to laboratory personnel. The need for a rapid and safe means of diagnosing coccidioidomycosis prompted us to develop a real-time PCR assay to detect Coccidioides spp. directly from clinical specimens. Primers and fluorescent resonance energy transfer (FRET) probes were designed to target the internal transcribed spacer 2 region of Coccidioides. The assay's limit of detection is below 50 targets per reaction. An analysis of 40 Coccidioides sp. clinical isolates grown in culture demonstrated 100% sensitivity of the assay. A cross-reactivity panel containing fungi, bacteria, mycobacteria, and viruses was tested and demonstrated 100% specificity for Coccidioides spp. An analysis of 266 respiratory specimens by LightCycler PCR demonstrated 100% sensitivity and 98.4% specificity for Coccidioides spp. compared with culture. Analysis of 66 fresh tissue specimens yielded 92.9% sensitivity and 98.1% specificity versus those of the culture method. The sensitivity of the assay testing 148 paraffin-embedded tissue samples is 73.4%. A rapid method for the detection of Coccidioides spp. directly from clinical material will greatly assist in the timely diagnosis and treatment of patients, while at the same time decreasing the risk of accidental exposure to laboratory personnel. PMID- 17108078 TI - Detection of multidrug resistance in Mycobacterium tuberculosis. AB - We developed a DNA sequencing-based method to detect mutations in the genome of drug-resistant Mycobacterium tuberculosis. Drug resistance in M. tuberculosis is caused by mutations in restricted regions of the genome. Eight genome regions associated with drug resistance, including rpoB for rifampin (RIF), katG and the mabA (fabG1)-inhA promoter for isoniazid (INH), embB for ethambutol (EMB), pncA for pyrazinamide (PZA), rpsL and rrs for streptomycin (STR), and gyrA for levofloxacin, were amplified simultaneously by PCR, and the DNA sequences were determined. It took 6.5 h to complete all procedures. Among the 138 clinical isolates tested, 55 were resistant to at least one drug. Thirty-four of 38 INH resistant isolates (89.5%), 28 of 28 RIF-resistant isolates (100%), 15 of 18 EMB resistant isolates (83.3%), 18 of 30 STR-resistant isolates (60%), and 17 of 17 PZA-resistant isolates (100%) had mutations related to specific drug resistance. Eighteen of these mutations had not been reported previously. These novel mutations include one in rpoB, eight in katG, one in the mabA-inhA regulatory region, two in embB, five in pncA, and one in rrs. Escherichia coli isolates expressing individually five of the eight katG mutations showed loss of catalase and INH oxidation activities, and isolates carrying any of the five pncA mutations showed no pyrazinamidase activity, indicating that these mutations are associated with INH and PZA resistance, respectively. Our sequencing-based method was also useful for testing sputa from tuberculosis patients and for screening of mutations in Mycobacterium bovis. In conclusion, our new method is useful for rapid detection of multiple-drug-resistant M. tuberculosis and for identifying novel mutations in drug-resistant M. tuberculosis. PMID- 17108079 TI - A 50-kDa ERK-like protein is up-regulated by a dual altered peptide ligand that suppresses myasthenia gravis-associated responses. AB - Myasthenia gravis (MG) and its animal model, experimental autoimmune MG (EAMG), are T cell-dependent antibody-mediated autoimmune diseases. A dual altered peptide ligand (APL) that is composed of the tandemly arranged two single amino acid analogues of two myasthenogenic peptides, p195-212 and p259-271, down regulated in vitro and in vivo MG-associated autoreactive responses. The dual APL was shown to exert its beneficial effects by up-regulating ERK1,2 in CD4(+)CD25(+) regulatory cells. In this study, we investigated a novel 50-kDa ERK like protein (ERK-50) that is up-regulated significantly in addition to ERK1,2 after treatment with the dual APL. We report here that ERK-50 was up-regulated in LN cells and in LN-derived T cells of mice that were immunized with the myasthenogenic peptides and treated with the dual APL. Moreover, ERK-50 was up regulated in dual-APL- treated mice that were immunized with the Torpedo acetylcholine receptor. ERK-50 was demonstrated to be recognized by antibodies directed against the C and N termini of ERK1, against the C terminus of ERK2, and against general ERK. The 50-kDa ERK was shown to be stimulated by Con A, and inhibition of MEK1 down-regulated the 50-kDa ERK as was shown for ERK1,2. However, 4beta-phorbol 12-myristate 13-acetate (TPA) did not stimulate ERK-50. Finally, the activated ERK-50 was up-regulated in the dual-APL-induced CD4(+)CD25(+) regulatory cells. Thus, ERK-50 is suggested to be a novel ERK isoform, being up-regulated in response to treatment with the dual APL. PMID- 17108080 TI - A signature pattern of stress-responsive microRNAs that can evoke cardiac hypertrophy and heart failure. AB - Diverse forms of injury and stress evoke a hypertrophic growth response in adult cardiac myocytes, which is characterized by an increase in cell size, enhanced protein synthesis, assembly of sarcomeres, and reactivation of fetal genes, often culminating in heart failure and sudden death. Given the emerging roles of microRNAs (miRNAs) in modulation of cellular phenotypes, we searched for miRNAs that were regulated during cardiac hypertrophy and heart failure. We describe >12 miRNAs that are up- or down-regulated in cardiac tissue from mice in response to transverse aortic constriction or expression of activated calcineurin, stimuli that induce pathological cardiac remodeling. Many of these miRNAs were similarly regulated in failing human hearts. Forced overexpression of stress-inducible miRNAs was sufficient to induce hypertrophy in cultured cardiomyocytes. Similarly, cardiac overexpression of miR-195, which was up-regulated during cardiac hypertrophy, resulted in pathological cardiac growth and heart failure in transgenic mice. These findings reveal an important role for specific miRNAs in the control of hypertrophic growth and chamber remodeling of the heart in response to pathological signaling and point to miRNAs as potential therapeutic targets in heart disease. PMID- 17108081 TI - Drosophila flies in "Evolution Canyon" as a model for incipient sympatric speciation. AB - The genetic basis of population divergence leading to adaptive radiation and speciation is a major unresolved problem of evolutionary biology. Molecular elucidation of "speciation genes" advanced recently, yet it remains without clear identification of the gene complexes participating in reproductive isolation between natural populations, particularly, in sympatry. Genetic divergence was discovered between Drosophila melanogaster populations inhabiting ecologically contrasting, opposite slopes in "Evolution Canyon" (EC), Mt. Carmel, Israel. Interslope migration of flies is easy and verified. Nevertheless, significant interslope D. melanogaster population divergence was established at EC involving habitat choice, mate choice, thermal and drought tolerances, adaptive genes, and mobile elements. Parallel patterns of stress tolerance, habitat choice, and mate choice were demonstrated in Drosophila simulans at EC, although on a smaller scale. However, some tests for interslope genetic differentiation in Drosophila, derived from the opposite EC slopes, gave somewhat controversial results. Here we present new empirical data on interslope genetic divergence of Drosophila at EC, and summarize previous supporting and controversial results. We suggest that Drosophila populations at EC represent a rare example, demonstrating how selection overrides migration, and propose an ad hoc ecological model of incipient sympatric divergence. PMID- 17108082 TI - Enhanced anxiety and stress-induced corticosterone release are associated with increased Crh expression in a mouse model of Rett syndrome. AB - Rett syndrome (RTT), a postnatal neurodevelopmental disorder, is caused by mutations in the methyl-CpG-binding protein 2 (MECP2) gene. Children with RTT display cognitive and motor abnormalities as well as autistic features. We studied mice bearing a truncated Mecp2 allele (Mecp2(308/Y) mice) and found evidence of increased anxiety-like behavior and an abnormal stress response as evidenced by elevated serum corticosterone levels. We found increased corticotropin-releasing hormone (Crh) gene expression in the paraventricular nucleus of the hypothalamus, the central amygdala, and the bed nucleus of the stria terminalis. Finally, we discovered that MeCP2 binds the Crh promoter, which is enriched for methylated CpG dinucleotides. In contrast, the MeCP2(308) protein was not detected at the Crh promoter. This study identifies Crh as a target of MeCP2 and implicates Crh overexpression in the development of specific features of the Mecp2(308/Y) mouse, thereby providing opportunities for clinical investigation and therapeutic intervention in RTT. PMID- 17108083 TI - Human SHPRH is a ubiquitin ligase for Mms2-Ubc13-dependent polyubiquitylation of proliferating cell nuclear antigen. AB - Human SHPRH gene is located at the 6q24 chromosomal region, and loss of heterozygosity in this region is seen in a wide variety of cancers. SHPRH is a member of the SWI/SNF family of ATPases/helicases, and it possesses a C(3)HC(4) RING motif characteristic of ubiquitin ligase proteins. In both of these features, SHPRH resembles the yeast Rad5 protein, which, together with Mms2 Ubc13, promotes replication through DNA lesions via an error-free postreplicational repair pathway. Genetic evidence in yeast has indicated a role for Rad5 as a ubiquitin ligase in mediating the Mms2-Ubc13-dependent polyubiquitylation of proliferating cell nuclear antigen. Here we show that SHPRH is a functional homolog of Rad5. Similar to Rad5, SHPRH physically interacts with the Rad6-Rad18 and Mms2-Ubc13 complexes, and we show that SHPRH protein is a ubiquitin ligase indispensable for Mms2-Ubc13-dependent polyubiquitylation of proliferating cell nuclear antigen. Based on these observations, we predict a role for SHPRH in promoting error-free replication through DNA lesions. Such a role for SHPRH is consistent with the observation that this gene is mutated in a number of cancer cell lines, including those from melanomas and ovarian cancers, which raises the strong possibility that SHPRH function is an important deterrent to mutagenesis and carcinogenesis in humans. PMID- 17108084 TI - 3D structure of amyloid protofilaments of beta2-microglobulin fragment probed by solid-state NMR. AB - Understanding the structure and formation of amyloid fibrils, the filamentous aggregates of proteins and peptides, is crucial in preventing diseases caused by their deposition and, moreover, for obtaining further insight into the mechanism of protein folding and misfolding. We have combined solid-state NMR, x-ray fiber diffraction, and atomic force microscopy to reveal the 3D structure of amyloid protofilament-like fibrils formed by a 22-residue K3 peptide (Ser(20)-Lys(41)) of beta(2)-microglobulin, a protein responsible for dialysis-related amyloidosis. Although a uniformly (13)C,(15)N-labeled sample was used for the NMR measurements, we could obtain the 3D structure of the fibrils on the basis of a large number of structural constraints. The conformation of K3 fibrils was found to be a beta-strand-loop-beta-strand with each K3 molecule stacked in a parallel and staggered manner. It is suggested that the fibrillar conformation is stabilized by intermolecular interactions, rather than by intramolecular hydrophobic packing as seen in globular proteins. Together with thermodynamic studies of the full-length protein, formation of the fibrils is likely to require side chains on the intermolecular surface to pack tightly against those of adjacent monomers. By revealing the structure of beta(2)-microglobulin protofilament-like fibrils, this work represents technical progress in analyzing amyloid fibrils in general through solid-state NMR. PMID- 17108085 TI - Human cancers express a mutator phenotype. AB - Cancer cells contain numerous clonal mutations, i.e., mutations that are present in most or all malignant cells of a tumor and have presumably been selected because they confer a proliferative advantage. An important question is whether cancer cells also contain a large number of random mutations, i.e., randomly distributed unselected mutations that occur in only one or a few cells of a tumor. Such random mutations could contribute to the morphologic and functional heterogeneity of cancers and include mutations that confer resistance to therapy. We have postulated that malignant cells exhibit a mutator phenotype resulting in the generation of random mutations throughout the genome. We have recently developed an assay to quantify random mutations in human tissue with unprecedented sensitivity. Here, we report measurements of random single nucleotide substitutions in normal and neoplastic human tissues. In normal tissues, the frequency of spontaneous random mutations is exceedingly low, less than 1 x 10(-8) per base pair. In contrast, tumors from the same individuals exhibited an average frequency of 210 x 10(-8) per base pair, an elevation of at least two orders of magnitude. Our data document tumor heterogeneity at the single-nucleotide level, indicate that accelerated mutagenesis prevails late into tumor progression, and suggest that elevation of random mutation frequency in tumors might serve as a novel prognostic indicator. PMID- 17108086 TI - Apparent cooperativity in the folding of multidomain proteins depends on the relative rates of folding of the constituent domains. AB - Approximately 75% of eukaryotic proteins contain more than one so-called independently folding domain. However, there have been relatively few systematic studies to investigate the effect of interdomain interactions on protein stability and fewer still on folding kinetics. We present the folding of pairs of three-helix bundle spectrin domains as a paradigm to indicate how complex such an analysis can be. Equilibrium studies show an increase in denaturant concentration required to unfold the domains with only a single unfolding transition; however, in some cases, this is not accompanied by the increase in m value, which would be expected if the protein is a truly cooperative, all-or-none system. We analyze the complex kinetics of spectrin domain pairs, both wild-type and carefully selected mutants. By comparing these pairs, we are able to demonstrate that equilibrium data alone are insufficient to describe the folding of multidomain proteins and to quantify the effects that one domain can have on its neighbor. PMID- 17108087 TI - Differential slow inactivation and use-dependent inhibition of Nav1.8 channels contribute to distinct firing properties in IB4+ and IB4- DRG neurons. AB - Nociceptive dorsal root ganglion (DRG) neurons can be classified into nonpeptidergic IB(4)(+) and peptidergic IB(4)(-) subtypes, which terminate in different layers in dorsal horn and transmit pain along different ascending pathways, and display different firing properties. Voltage-gated, tetrodotoxin resistant (TTX-R) Na(v)1.8 channels are expressed in both IB(4)(+) and IB(4)(-) cells and produce most of the current underlying the depolarizing phase of action potential (AP). Slow inactivation of TTX-R channels has been shown to regulate repetitive DRG neuron firing behavior. We show in this study that use-dependent reduction of Na(v)1.8 current in IB(4)(+) neurons is significantly stronger than that in IB(4)(-) neurons, although voltage dependency of activation and steady state inactivation are not different. The time constant for entry of Na(v)1.8 into slow inactivation in IB(4)(+) neurons is significantly faster and more Na(v)1.8 enter the slow inactivation state than in IB(4)(-) neurons. In addition, recovery from slow inactivation of Na(v)1.8 in IB(4)(+) neurons is slower than that in IB(4)(-) neurons. Using current-clamp recording, we demonstrate a significantly higher current threshold for generation of APs and a longer latency to onset of firing in IB(4)(+), compared with those of IB(4)(-) neurons. In response to a ramp stimulus, IB(4)(+) neurons produce fewer APs and display stronger adaptation, with a faster decline of AP peak than IB(4)(-) neurons. Our data suggest that differential use-dependent reduction of Na(v)1.8 current in these two DRG subpopulations, which results from their different rate of entry into and recovery from the slow inactivation state, contributes to functional differences between these two neuronal populations. PMID- 17108088 TI - Spatial summation, end inhibition and side inhibition in the middle temporal visual area (MT). AB - We investigated the responses of single neurons in the middle temporal area (MT) of anesthetized marmoset monkeys to sine-wave gratings of various lengths and widths. For the vast majority of MT cells maximal responses were obtained on presentation of gratings of specific dimensions, which were typically asymmetrical along the length and width axes. The strength of end inhibition was dependent on the width of the stimulus, with many cells showing clear end inhibition only when wide gratings were used. Conversely, the strength of side inhibition was dependent on stimulus length. Furthermore, for over one third of MT cells length summation properties could not be defined without consideration of stimulus width and vice versa. These neurons, which we refer to as "length width inseparable" (LWI) cells, were rare in layer 4. The majority of LWI neurons was strongly inhibited by wide-field stimuli and responded preferentially to gratings that were elongated, along either the length or width dimensions. However, rather than forming a homogeneous and entirely distinct group, LWI cells represented the upper end of a continuum of complexity in spatial summation response properties, which characterized the population of MT cells. Only a minority of MT neurons (22.3%) showed no evidence of inhibition by wide-field stimuli, with this type of response being common among layer 5 cells. These results demonstrate distinct patterns of spatial selectivity in MT, supporting the notion that neurons in this area can perform various roles in terms of grouping and segmentation of motion signals. PMID- 17108089 TI - Increased nociceptive input rapidly modulates spinal GABAergic transmission through endogenously released glutamate. AB - Stimulation of nociceptive primary afferents elicits pain by promoting glutamatergic transmission in the spinal cord. Little is known about how increased nociceptive input controls GABAergic tone in the spinal dorsal horn. In this study, we determined how increased nociceptive inflow affects GABAergic spontaneous inhibitory postsynaptic currents (sIPSCs) of lamina II neurons by using whole cell recordings in rat spinal cord slices. Bath application of capsaicin for 3 min induced a long-lasting inhibition of sIPSCs in 50% of the neurons tested. In the other half of the neurons, capsaicin either increased the frequency of sIPSCs (34.6%) or had no effect on sIPSCs (15.4%). The GABA(A) current elicited by puff application of GABA was not altered by capsaicin. Capsaicin did not inhibit sIPSCs in rats treated with intrathecal pertussis toxin. Also, capsaicin failed to inhibit sIPSCs in the presence of ionotropic glutamate receptor antagonists or in the presence of both LY341495 and CPPG (group II and group III metabotropic glutamate receptor antagonists, respectively). However, when LY341495 or CPPG was used alone, capsaicin still decreased the frequency of sIPSCs in some neurons. Additionally, bradykinin significantly inhibited sIPSCs in a population of lamina II neurons and this inhibitory effect was also abolished by LY341495 and CPPG. Our study provides novel information that stimulation of nociceptive primary afferents rapidly suppresses GABAergic input to many dorsal horn neurons through endogenous glutamate and activation of presynaptic group II and group III metabotropic glutamate receptors. These findings extend our understanding of the microcircuitry of the spinal dorsal horn involved in nociception. PMID- 17108090 TI - Long-lasting, context-dependent modification of stepping in the cat after repeated stumbling-corrective responses. AB - A consistent feature of animal locomotion is the capacity to maintain stable movements in changing environments. Here we describe long-term modification of the swing movement of the hind leg in cats in response to repeatedly impeding the movement of the leg. While studying phase transitions in the hind legs, we discovered that repetitively evoking the stumbling-corrective reaction led to long-lasting increases in knee flexion and step height during swing to avoid the impediment. These increases were apparent after nearly 20 stimuli and maximal after about 120 stimuli and, in some animals, they persisted for > or =24 h after presentation of the stimuli. Furthermore, these long-lasting changes were context dependent and did not generalize to other environments; when walking was observed in an environment distinct from that used in training, the hind-limb kinematics returned to normal. To gain insight into what regions of the nervous system might be involved in this long-term modification, we examined the changes in stepping in decerebrate cats after multiple perturbed steps. In this situation, there was a short-term increase in step height, although this increase was smaller than that evoked in intact animals and persisted for <1 min after termination of the stimuli. Thus induction of the long-term increase in step height requires the forebrain. We propose that the conditioned change in leg movement is related to a general ability of animals to adapt locomotor movements to new features of the environment. PMID- 17108091 TI - Visual-vestibular interaction hypothesis for the control of orienting gaze shifts by brain stem omnipause neurons. AB - Models of combined eye-head gaze shifts all aim to realistically simulate behaviorally observed movement dynamics. One of the most problematic features of such models is their inability to determine when a saccadic gaze shift should be initiated and when it should be ended. This is commonly referred to as the switching mechanism mediated by omni-directional pause neurons (OPNs) in the brain stem. Proposed switching strategies implemented in existing gaze control models all rely on a sensory error between instantaneous gaze position and the spatial target. Accordingly, gaze saccades are initiated after presentation of an eccentric visual target and subsequently terminated when an internal estimate of gaze position becomes nearly equal to that of the target. Based on behavioral observations, we demonstrate that such a switching mechanism is insufficient and is unable to explain certain types of movements. We propose an improved hypothesis for how the OPNs control gaze shifts based on a visual-vestibular interaction of signals known to be carried on anatomical projections to the OPN area. The approach is justified by the analysis of recorded gaze shifts interrupted by a head brake in animal subjects and is demonstrated by implementing the switching mechanism in an anatomically based gaze control model. Simulated performance reveals that a weighted sum of three signals: gaze motor error, head velocity, and eye velocity, hypothesized as inputs to OPNs, successfully reproduces diverse behaviorally observed eye-head movements that no other existing model can account for. PMID- 17108092 TI - Contrast affects speed tuning, space-time slant, and receptive-field organization of simple cells in macaque V1. AB - We measured speed tuning of V1 cells in alert macaques to high- and low-contrast stimuli. Most V1 cells tested, both simple and complex and directional as well as nondirectional, shifted their speed tuning to slower speeds for lower contrast stimuli. We found that the space-time slant of the receptive field of directional simple cells differed for high- and low-contrast stimuli, with the space-time slant predicting higher optimum speeds for the higher-contrast stimuli; i.e., there was a larger spatial shift of the receptive-field organization per unit time. Not only did the space-time maps of directional simple cells show different slants between high- and low-contrast stimuli, but they also showed a different organization, because for high-contrast stimuli, the maps tended to show a complete inversion of the receptive-field spatial organization at long delays after stimulus onset, with initial excitation followed by suppression and initial suppression followed by excitation, but for low-contrast stimuli the receptive field organization showed only a quadrature shift over time. We show that a simple modification of earlier models for the generation of direction-selective simple cells can account for these observations. PMID- 17108093 TI - Developmental changes in retrograde messengers involved in depolarization-induced suppression of excitation at parallel fiber-Purkinje cell synapses in rodents. AB - At parallel fiber (PF) to Purkinje cell (PC) synapses, depolarization-induced suppression of excitation (DSE) and suppression of PF-excitatory postsynaptic currents (EPSCs) by activation of postsynaptic mGluR1 glutamate (Glu) receptors involve retrograde release of endocannabinoids. However, Levenes et al. suggested instead that Glu was the retrograde messenger in this latter case. Because the study by Levenes et al. was performed in nearly mature rats, whereas most others were performed in juvenile animals, DSE was re-investigated in juvenile versus nearly mature rats and mice. Indeed, DSE was preferred here to agonist-induced suppression of PF-EPSCs, to avoid possible indirect effects in this latter case. In 10- to 12-day-old rats, DSE of PF-EPSCs was entirely mediated through retrograde release of endocannabinoids. In 18- to 22-day-old-rats, DSE was partly resistant to CB1 cannabinoid receptor antagonists. The remaining component was potentiated by the Glu uptake inhibitor d-threo-beta-benzyloxyaspartate (d-TBOA) and blocked by the desensitizing kainate (KA) receptor agonist (2S,4R)-4 methylglutamic acid (SYM 2081). This SYM-2081-sensitive component of DSE was accompanied by a paired-pulse facilitation increase that was also potentiated by d-TBOA and blocked by SYM 2081. In nearly mature wild-type and GluR6 -/- mice, results fully confirmed the presence of an endocannabinoid-independent component of DSE that involves retrograde release of Glu and activation of presynaptic KA receptors including GluR6 receptor subunits. Therefore retrograde release of Glu by PCs participates to DSE at PF-PC synapses in nearly mature rodents but not in juvenile ones, and Glu probably operates through activation of presynaptic KA receptors that include GluR6 receptor subunits. PMID- 17108094 TI - A population coding account for systematic variation in saccadic dead time. AB - During movement programming, there is a point in time at which the movement system is committed to executing an action with certain parameters even though new information may render this action obsolete. For saccades programmed to a visual target this period is termed the dead time. Using a double-step paradigm, we examined potential variability in the dead time with variations in overall saccade latency and spatiotemporal configuration of two sequential targets. In experiment 1, we varied overall saccade latency by manipulating the presence or absence of a central fixation point. Despite a large and robust gap effect, decreasing the saccade latency in this way did not alter the dead time. In experiment 2, we varied the separation between the two targets. The dead time increased with separation up to a point and then leveled off. A stochastic accumulator model of the oculomotor decision mechanism accounts comprehensively for our findings. The model predicts a gap effect through changes in baseline activity without producing variations in the dead time. Variations in dead time with separation between the two target locations are a natural consequence of the population coding assumption in the model. PMID- 17108095 TI - Cortical response to auditory motion suggests an asymmetry in the reliance on inter-hemispheric connections between the left and right auditory cortices. AB - The aim of the current study was to measure the brain's response to auditory motion using electroencephalography (EEG) to gain insight into the mechanisms by which hemispheric lateralization for auditory spatial processing is established in the human brain. The onset of left- or rightward motion in an otherwise continuous sound was found to elicit a large response, which appeared to arise from higher-level nonprimary auditory areas. This motion onset response was strongly lateralized to the hemisphere contralateral to the direction of motion. The response latencies suggest that the ipsilateral response to the leftward motion was produced by indirect callosal projections from the opposite hemisphere, whereas the ipsilateral response to the rightward motion seemed to receive contributions from direct thalamocortical projections. These results suggest an asymmetry in the reliance on inter-hemispheric projections between the left and right auditory cortices for auditory spatial processing. PMID- 17108096 TI - Template-based spike pattern identification with linear convolution and dynamic time warping. AB - Pattern identification for spiking activity, which is central to neurophysiological analysis, is complicated by variability in spiking at multiple timescales. Incorporating likelihood tests on the variability at two timescales, we developed an approach to identifying segments from continuous neurophysiological recordings that match preselected spike "templates." At smaller timescales, each component of the preselected pattern is represented by a linear filter. Local scores to measure the similarities between short data segments and the pattern components are computed as filter responses. At larger timescales, overall scores to measure the similarities between relatively long data segments and the entire pattern are computed by dynamic time warping, which combines the local similarity scores associated with the pattern components, optimizing over a range of intercomponent time intervals. Occurrences of the pattern are identified by local peaks in the overall similarity scores. This approach is developed for point process representations and binary representations of spiking activity, both deriving from a single underlying statistical model. Point process representations are suitable for highly reliable single-unit responses, whereas binary representations are preferred for more variable single-unit responses and multiunit responses. Testing with single units recorded from individual electrodes within the robust nucleus of the arcopallium of zebra finches and with recordings from an array placed within the motor cortex of macaque monkeys demonstrates that the approach can identify occurrences of specified patterns with good time precision in a broad range of neurophysiological data. PMID- 17108097 TI - Enhancement of object representations in primate perirhinal cortex during a visual working-memory task. AB - We compared single-cell activities in perirhinal cortex (PRh) as well as adjacent visual cortex (area TE) across two tasks. One task required the monkey to identify any stimulus repetition within a sequence of object stimuli. In the other task, the same stimuli were presented, but the monkey didn't have to remember them. PRh responses during the object-memory task were elevated relative to those during the second task. In TE, on the other hand, there were no significant task-related differences in responses. We did not observe task related differences related to repetition effects in either brain area. The onset of the enhanced signal in PRh during the object-memory task occurred with a latency of 80 ms after the onset of the stimulus response, suggesting that it was the result of top-down feedback. PMID- 17108098 TI - A causal role for the human tumor antigen preferentially expressed antigen of melanoma in cancer. AB - Tumor antigens are of interest as diagnostic and prognostic markers and potential therapeutic targets. The tumor antigen preferentially expressed antigen of melanoma (PRAME) is frequently overexpressed in a wide variety of cancers and is a prognostic marker for clinical outcome. It has been shown recently that PRAME functions as a repressor of retinoic acid signaling. Here, we discuss this novel insight in the context of the increasing interest in tumor antigens as targets for therapy. PMID- 17108099 TI - Oncogenes, trousseau syndrome, and cancer-related changes in the coagulome of mice and humans. AB - Cancer is often associated with venous thrombosis, a phenomenon that was first described by Trousseau in 1865 (Trousseau syndrome). Recent studies have begun to explain how oncogenic events may deregulate the hemostatic system. For instance, activated oncogenes (K-ras, EGFR, PML-RARalpha, and MET) or inactivated tumor suppressors (e.g., 53 or PTEN) may increase the risk of thrombosis by inducing the expression of tissue factor, a potent procoagulant molecule, and plasminogen activator inhibitor-1, a fibrinolysis inhibitor. In a more complex clinical reality, transforming genes may often act in concert with numerous epigenetic factors, including hypoxia, inflammation, anticancer therapy, contact between blood and metastatic cancer cells, and emission of procoagulant vesicles from tumors and their stroma into the circulation. To add to mechanistic insights gained from mouse models, which may not fully phenocopy human Trousseau syndrome, we suggest that valuable clues to progression and thrombosis risk may be obtained by monitoring multiple hemostatic variables in cancer patients ("coagulomics"). PMID- 17108100 TI - Caveolin-1, mammary stem cells, and estrogen-dependent breast cancers. AB - Estrogen exposure is considered a significant risk factor for breast cancer development. Estrogen receptor (ER) alpha is expressed at low levels in normal epithelia, and its expression is dramatically up-regulated as transformation progresses during mammary hyperplasia and adenocarcinoma development. The mechanism(s) driving ERalpha up-regulation during mammary tumorigenesis remains unclear. Caveolin-1 (Cav-1) is the structural protein of plasmalemmal invaginations, termed caveolae, which functions as a tumor suppressor gene. Interestingly, Cav-1 dominant-negative mutations are exclusively found in ERalpha positive breast cancer samples. In support of these clinical findings, ERalpha expression is increased in Cav-1 (-/-) null mammary epithelia, and estrogen stimulation further enhances the growth of Cav-1-deficient three-dimensional epithelial structures. These phenotypes correlate with augmented levels of cyclin D1. In addition, Cav-1 gene inactivation induces the accumulation of a cell population with the characteristics of adult mammary stem cells. Primary cultures of Cav-1 (-/-) mammary epithelial cells exhibit premalignant changes, such as abnormal lumen formation, epidermal growth factor-independent growth, defects in cell substrate attachment, and increased cell invasiveness. Thus, Cav-1 gene inactivation promotes premalignant alterations in mammary epithelia and induces increased ERalpha expression levels and the up-regulation of cyclin D1. As tumor formation is a multihit process, Cav-1 mutations that occur during the early stages of mammary transformation may be a critical upstream/initiating event leading to increased ERalpha levels. PMID- 17108101 TI - Markers and tissue resources for melanoma: meeting report. AB - The Markers and Tissue Resources for Melanoma meeting convened by the Cancer Diagnosis Program, Division of Cancer Treatment and Diagnosis, Specialized Programs of Research Excellence at the Organ Systems Branch of the National Cancer Institute (NCI), and the Melanoma Research Foundation was held in Gaithersburg, MD on October 2005. The meeting reviewed the current status of biomarkers for early- and advanced-stage melanoma and addressed some of the challenges scientists and clinicians face as they unravel the biology of melanoma and try to apply these findings to patient care. Specifically, the participants focused on molecular changes associated with melanoma progression, potential diagnostic and prognostic markers emerging from molecular profiling studies, and new treatment targets for current and future clinical trials. They also highlighted the ongoing challenges about translational research in melanoma, including availability of tissue resources, and summarized the status of nevus and melanoma tissue microarrays, recently developed as a collaborative project between the melanoma research community and the NCI. The meeting report is intended to provide a perspective on emerging scientific approaches in translational research that can enhance the progress in discovery and validation of markers for melanoma. (Cancer Res 2006; 66(22): 10652-7). PMID- 17108102 TI - TMPRSS2:ERG fusion by translocation or interstitial deletion is highly relevant in androgen-dependent prostate cancer, but is bypassed in late-stage androgen receptor-negative prostate cancer. AB - Recently, a unique fusion between the prostate-specific, androgen-regulated TMPRSS2 gene and the ETS genes ERG, ETV1, or ETV4 has been described in clinical prostate cancer. We investigated mechanisms of expression of four ETS genes, ERG, ETV1, ETV4, and FLI1, in 11 xenografts representing different stages of prostate cancer. All five androgen-dependent xenografts showed as major transcript overexpression of two splice variants of TMPRSS2:ERG, linking TMPRSS2 exon 1 or 2 sequences to ERG exon 4. In one of two androgen-sensitive xenografts, fusion transcripts of TMPRSS2 and ETV1 were detected. Array-based comparative genomic hybridization and interphase fluorescence in situ hybridization indicated both interstitial deletions and translocations as mechanisms of TMPRSS2:ERG gene fusion. Importantly, TMPRSS2 to ERG fusions were also observed in three of four androgen-independent, androgen receptor (AR)-negative xenografts and in two AR negative clinical prostate cancer specimens; however, the fusion gene was not expressed. In almost all AR-negative tumor samples, overexpression of wild-type ETV4 or FLI1 was detected. Combined, our observations indicate a key role of fusion of TMPRSS2 and ETS genes in most androgen-regulated prostate cancers, which might be bypassed by androgen-independent expression of wild-type ETS factors in late-stage disease. PMID- 17108103 TI - p53 Attenuates cancer cell migration and invasion through repression of SDF 1/CXCL12 expression in stromal fibroblasts. AB - The p53 tumor suppressor acts as a major barrier against cancer. To a large extent, this is due to its ability to maintain genome stability and to eliminate cancer cells from the replicative pool through cell-autonomous mechanisms. However, in addition to its well-documented functions within the malignant cancer cell, p53 can also exert non-cell-autonomous effects that contribute to tumor suppression. We now report that p53 can suppress the production of the chemokine SDF-1 in cultured fibroblasts of both human and mouse origin. This is due to a p53-mediated down-regulation of SDF-1 mRNA, which can be exacerbated on activation of p53 by the drug Nutlin-3. SDF-1 promotes the migration and invasiveness of cells that express its cognate receptor CXCR4. Indeed, medium conditioned by p53-deficient fibroblasts induces cancer cells towards increased directional migration and invasiveness, which are largely reversed by CXCR4 antagonist peptides. Because SDF-1 produced by stromal fibroblasts plays an important role in cancer progression and metastasis, our findings suggest that the ability of p53 to suppress stromal SDF-1 production may be an important mechanism whereby it does its non-cell-autonomous tumor suppressor function. PMID- 17108104 TI - Human arrest defective 1 acetylates and activates beta-catenin, promoting lung cancer cell proliferation. AB - Arrest defective 1 (ARD1), an acetyltransferase, is essential for the yeast life cycle. Although its human homologue (hARD1) has been identified, its biological functions in human cells remain unclear. In the present study, we examined the biological function of hARD1. In H1299 and A549 lung cancer cells, hARD1 silencing RNA inhibited cell proliferation and induced G(1) arrest. Cyclin D1 was also found to be down-regulated in these growth-arrested cells, and the ectopic expression of cyclin D1 rescued cell growth. hARD1 knockdown repressed the promoter activity of the cyclin D1 gene, which inhibited the transcription of cyclin D1. Moreover, hARD1 knockdown reduced the binding of beta-catenin/TCF4 transcription factor to cyclin D1 promoter and repressed its transcriptional activity. Inversely, hARD1 expression increased the transcriptional activity of beta-catenin. Both endogenous and ectopically expressed hARD1 was coimmunoprecipitated with beta-catenin. hARD1 knockdown did not affect beta catenin expression or degradation but noticeably reduced acetylated beta-catenin. The beta-catenin binding and acetylation by hARD1 were observed in vitro. Therefore, it is suggested that hARD1 participates in proliferation of lung cancer cells via the activation of beta-catenin. PMID- 17108105 TI - Decreased NKX3.1 protein expression in focal prostatic atrophy, prostatic intraepithelial neoplasia, and adenocarcinoma: association with gleason score and chromosome 8p deletion. AB - NKX3.1 is a homeobox gene located at chromosome 8p21.2, and one copy is frequently deleted in prostate carcinoma. Prior studies of NKX3.1 mRNA and protein in human prostate cancer and prostatic intraepithelial neoplasia (PIN) have been conflicting, and expression in focal prostate atrophy lesions has not been investigated. Immunohistochemical staining for NKX3.1 on human tissue microarrays was decreased in most focal atrophy and PIN lesions. In carcinoma, staining was inversely correlated with Gleason grade. Fluorescence in situ hybridization showed that no cases of atrophy had loss or gain of 8p, 8 centromere, or 8q24 (C-MYC) and only 12% of high-grade PIN lesions harbored loss of 8p. By contrast, NKX3.1 staining in carcinoma was correlated with 8p loss and allelic loss was inversely related to Gleason pattern. Quantitative reverse transcription-PCR for NKX3.1 mRNA using microdissected atrophy revealed a concordance with protein in five of seven cases. In carcinoma, mRNA levels were decreased in 6 of 12 cases but mRNA levels correlated with protein levels in only 4 of 12 cases, indicating translational or post-translational control. In summary, NKX3.1 protein is reduced in focal atrophy and PIN but is not related to 8p allelic loss in these lesions. Therefore, whereas genetic disruption of NKX3.1 in mice leads to PIN, nongenetic mechanisms reduce NKX3.1 protein levels early in human prostate carcinogenesis, which may facilitate both proliferation and DNA damage in atrophic and PIN cells. Monoallelic deletions on chromosome 8p are associated with more advanced invasive and aggressive disease. PMID- 17108106 TI - Telomere-driven karyotypic complexity concurs with p16INK4a inactivation in TP53 competent immortal endothelial cells. AB - Critically short telomeres promote chromosomal fusions, which in TP53-defective cells initiate the formation of cytogenetic aberrations that are typical of human cancer cells. Expression of the enzyme telomerase stabilizes normal and aberrant chromosomes by maintaining telomere length. However, previous investigations, including our own, have shown that overexpression of telomerase reverse transcriptase (hTERT) does not prevent net telomere shortening in human endothelial cells. In the present study, two mass cultures of hTERT-transduced bone marrow endothelial cells (BMhTERT) and 26 clones were employed to further investigate the immortalization process and consequences of telomere shortening. Eighty-five percent (22 of 26) of the clones and both mass cultures were immortalized. However, cytogenetic analyses revealed recurring cytogenetic aberrations in the mass cultures and 12 representative clones. Several of the recurring aberrations, including +5p, +11, -13, +19, and +20, and nonreciprocal translocations involving 17p and 2p were previously implicated in human carcinogenesis. One mass culture and a subset of clones (5 of 12) had complex karyotypes, characterized by cytogenetic heterogeneity and at least five chromosomal abnormalities. p16(INK4a) was silenced exclusively in the five clones and mass culture with complex karyotypes, whereas the p53/p21(cip1) pathway was defective in only one clone. Telomere dysfunction was implicated in the evolution of complex karyotypes by the presence of anaphase bridges, telomere associations, and dicentric chromosomes. These results show that complex karyotypes can evolve in TP53-competent cells and provide evidence that p16(INK4a) functions as a gatekeeper to prevent telomere-driven cytogenetic evolution. These investigations provide new insight to the role of p16(INK4a) as a tumor suppressor. PMID- 17108107 TI - LKB1 is recruited to the p21/WAF1 promoter by p53 to mediate transcriptional activation. AB - The tumor suppressor LKB1 is an evolutionarily conserved serine/threonine kinase. In humans, LKB1 can be inactivated either by germ-line mutations resulting in Peutz-Jeghers syndrome or by somatic mutations causing predisposition to multiple sporadic cancers. LKB1 has wide-ranging functions involved in tumor suppression and cell homeostasis, including establishing cell polarity, setting energy metabolic balance (via phosphorylation of AMP-dependent kinase), regulating the cell cycle, and promoting apoptosis. LKB1 function was previously linked to the tumor suppressor p53 and shown to activate the p53 target gene p21/WAF1. In this study, we further investigated LKB1 activation of the p21/WAF1 gene and addressed whether LKB1 is directly involved at the gene promoter. We find that, consistent with previous studies, LKB1 stabilizes p53 in vivo, correlating with activation of p21/WAF1. We show that LKB1 physically associates with p53 in the nucleus and directly or indirectly phosphorylates p53 Ser15 (previously shown to be phosphorylated by AMP-dependent kinase) and p53 Ser392. Further, these two p53 residues are required for LKB1-dependent cell cycle G(1) arrest. Chromatin immunoprecipitation analyses show that LKB1 is recruited directly to the p21/WAF1 promoter, as well as to other p53 activated promoters, in a p53-dependent fashion. Finally, a genetic fusion of LKB1 to defective p53, deleted for its activation domains, promotes activation of p21/WAF1. These results indicate that LKB1 has a direct role in activation of p21/WAF1 gene. PMID- 17108108 TI - Improved tumor control through circadian clock induction by Seliciclib, a cyclin dependent kinase inhibitor. AB - The circadian timing system and the cell division cycle are frequently deregulated in cancer. The therapeutic relevance of the reciprocal interactions between both biological rhythms was investigated using Seliciclib, a cyclin dependent kinase (CDK) inhibitor (CDKI). Mice bearing Glasgow osteosarcoma received Seliciclib (300 mg/kg/d orally) or vehicle for 5 days at Zeitgeber time (ZT) 3, 11, or 19. On day 6, tumor mRNA 24-hour expression patterns were determined for clock genes (Per2, Rev-erbalpha, and Bmal1) and clock-controlled cell cycle genes (c-Myc, Wee1, cyclin B1, and CDK1) with quantitative reverse transcription-PCR. Affinity chromatography on immobilized Seliciclib identified CDK1/CDK2 and extracellular signal-regulated kinase (ERK) 1/ERK2, CDK7/CDK9, and casein kinase CK1epsilon as Seliciclib targets, which respectively regulate cell cycle, transcription, and circadian clock in Glasgow osteosarcoma. Seliciclib reduced tumor growth by 55% following dosing at ZT3 or ZT11 and by 35% at ZT19 compared with controls (P < 0.001). Tolerability was also best at ZT3. Mean transcriptional activity of Rev-erbalpha, Per2, and Bmal1 was arrhythmic in the tumors of untreated mice. Seliciclib induced rhythmic clock gene expression patterns with physiologic phase relations only after ZT3 dosing. c-Myc and Wee1 mRNAs displayed synchronous circadian rhythms in the tumors of control mice receiving vehicle only but not in those of mice given the drug. Seliciclib further enhanced Wee1 expression irrespective of dosing time, an effect that reinforced G(2)-M gating. Seliciclib also inhibited CK1epsilon, which determines circadian period length. The coordination of clock gene expression patterns in tumor cells was associated with best antitumor activity of Seliciclib. The circadian clock and its upstream regulators represent relevant targets for CDKIs. PMID- 17108109 TI - Up-regulation of expression of the ubiquitin carboxyl-terminal hydrolase L1 gene in human airway epithelium of cigarette smokers. AB - Neuroendocrine differentiation is a common feature of lung cancer and increased numbers of neuroendocrine cells and their peptides have been described in chronic smokers. To understand the effects of cigarette smoking on the gene expression profile of neuroendocrine cells, microarray analysis with TaqMan confirmation was used to assess airway epithelial samples obtained by fiberoptic bronchoscopy from 81 individuals [normal nonsmokers, normal smokers, smokers with early chronic obstructive lung disease (COPD), and smokers with established COPD]. Of 11 genes considered to be neuroendocrine cell specific, only ubiquitin carboxyl-terminal hydrolase L1 (UCHL1), a member of the ubiquitin proteasome pathway, was consistently up-regulated in smokers compared with nonsmokers. Up-regulation of UCHL1 at the protein level was observed with immunohistochemical analysis of bronchial biopsies of smokers compared with nonsmokers. UCHL1 expression was evident only in neuroendocrine cells of the airway epithelium in nonsmokers; however, UCHL1 was also expressed in ciliated epithelial cells in smokers. This observation may add further weight to recent observations that ciliated cells are capable of transdifferentiating to other airway epithelial cells. In the context that UCHL1 is involved in the degradation of unwanted, misfolded, or damaged proteins within the cell and is overexpressed in >50% of lung cancers, its overexpression in chronic smokers may represent an early event in the complex transformation from normal epithelium to overt malignancy. PMID- 17108110 TI - Molecular mechanisms of RET receptor-mediated oncogenesis in multiple endocrine neoplasia 2B. AB - Multiple endocrine neoplasia 2B (MEN 2B) is an inherited syndrome of early onset endocrine tumors and developmental anomalies. The disease is caused primarily by a methionine to threonine substitution of residue 918 in the kinase domain of the RET receptor (2B-RET); however, the molecular mechanisms that lead to the disease phenotype are unclear. In this study, we show that the M918T mutation causes a 10 fold increase in ATP binding affinity and leads to a more stable receptor-ATP complex, relative to the wild-type receptor. Further, the M918T mutation alters local protein conformation, correlating with a partial loss of RET kinase autoinhibition. Finally, we show that 2B-RET can dimerize and become autophosphorylated in the absence of ligand stimulation. Our data suggest that multiple distinct but complementary molecular mechanisms underlie the MEN 2B phenotype and provide potential targets for effective therapeutics for this disease. PMID- 17108111 TI - Mutant p53 protects cells from 12-O-tetradecanoylphorbol-13-acetate-induced death by attenuating activating transcription factor 3 induction. AB - Mutations in p53 are ubiquitous in human tumors. Some p53 mutations not only result in loss of wild-type (WT) activity but also grant additional functions, termed "gain of function." In this study, we explore how the status of p53 affects the immediate response gene activating transcription factor 3 (ATF3) in the 12-O-tetradecanoylphorbol-13-acetate (TPA)-protein kinase C (PKC) pathway. We show that high doses of TPA induce ATF3 in a WT p53-independent manner correlating with PKCs depletion and cell death. We show that cells harboring mutant p53 have attenuated ATF3 induction and are less sensitive to TPA-induced death compared with their p53-null counterparts. Mutagenesis analysis of the ATF3 promoter identified the regulatory motifs cyclic AMP-responsive element binding protein/ATF and MEF2 as being responsible for the TPA-induced activation of ATF3. Moreover, we show that mutant p53 attenuates ATF3 expression by two complementary mechanisms. It interacts with the ATF3 promoter and influences its activity via the MEF2 site, and additionally, it attenuates transcriptional expression of the ATF3 activator MEF2D. These data provide important insights into the molecular mechanisms that underlie mutant p53 gain of function. PMID- 17108112 TI - Chronic protein kinase B (PKB/c-akt) activation leads to apoptosis induced by oxidative stress-mediated Foxo3a transcriptional up-regulation. AB - Increased protein kinase B (PKB; c-Akt) activation is a hallmark of diverse neoplasias providing both proliferative and antiapoptotic survival signals. In this study, we investigated the effect of chronic PKB activation on cellular survival and proliferation using cytokine-dependent bone marrow-derived Ba/F3 cells, in which PKBalpha activation can be directly, and specifically, induced by addition of 4-hydroxytamoxifen (4-OHT). Direct activation of PKB rescued Ba/F3 cells from cytokine withdrawal-induced apoptosis; however, surprisingly, these antiapoptotic effects were short lived, cells only being protected for up to 48 hours. We observed that activation of PKB in survival factor-deprived cells led to a dramatic increase of Foxo3a on both the transcriptional and protein level leading to expression of its transcriptional targets Bim and p27(kip1). High levels of PKB activity result in increased aerobic glycolysis and mitochondrial activity resulting in overproduction of reactive oxygen species. To determine whether oxidative stress might itself be responsible for Foxo3a up-regulation, we utilized hydrogen peroxide (H(2)O(2)) as an artificial inducer of oxidative stress and N-acetylcysteine (NAC), a thiol-containing radical oxygen scavenger. Addition of NAC to the culture medium prolonged the life span of cells treated with 4-OHT and prevented the up-regulation of Foxo3a protein levels caused by PKB activation. Conversely, treatment of Ba/F3 cells with H(2)O(2) caused an increase of Foxo3a on both transcriptional and protein levels, suggesting that deregulated PKB activation leads to oxidative stress resulting in Foxo3a up-regulation and subsequently cell death. Taken together, our data provide novel insights into the molecular consequences of uncontrolled PKB activation. PMID- 17108113 TI - Epigenetic regulation of tumor endothelial cell anergy: silencing of intercellular adhesion molecule-1 by histone modifications. AB - Tumors can escape from immunity by repressing leukocyte adhesion molecule expression on tumor endothelial cells and by rendering endothelial cells unresponsive to inflammatory activation. This endothelial cell anergy is induced by angiogenic growth factors and results in reduced leukocyte-vessel wall interactions, thereby attenuating infiltration of leukocytes into the tumor. This report describes a novel mechanism of endothelial cell anergy regulation. We recently reported that DNA methyltransferase (DNMT) and histone deacetylase (HDAC) inhibitors have angiostatic activity. Here, we studied whether epigenetic mechanisms regulate this angiogenesis-mediated escape from immunity. We found that DNMT inhibitors 5-aza-2'-deoxycytidine and zebularine, as well as HDAC inhibitor trichostatin A, reexpressed intercellular adhesion molecule-1 (ICAM-1) on tumor-conditioned endothelial cells in vitro, resulting in restored leukocyte endothelial cell adhesion. In addition, treatment with DNMT or HDAC inhibitors in vivo also restored ICAM-1 expression on tumor endothelial cells from two different mouse tumor models. Furthermore, leukocyte-vessel wall interactions in mouse tumors were increased by these compounds, as measured by intravital microscopy, resulting in enhanced leukocyte infiltration. We show that ICAM-1 down-regulation in tumor endothelial cells is associated with ICAM-1 promoter histone H3 deacetylation and loss of histone H3 Lys(4) methylation but not with DNA hypermethylation. In conclusion, our data show that ICAM-1 is epigenetically silenced in tumor endothelial cells by promoter histone modifications, which can be overcome by DNMT and HDAC inhibitors, suggesting a new molecular mechanism based on which novel therapeutic approaches for cancer can be pursued. PMID- 17108114 TI - Imaging mitogen-activated protein kinase function in xenograft models of prostate cancer. AB - Mitogen-activated protein kinases (MAPK) play important roles in malignancy. The ability to detect and quantitate MAPKs in live animal models of cancer will facilitate an understanding of disease progression. We have developed a gene expression-based imaging system that detects and quantifies MAPK activity in prostate cancer tumors implanted into severe combined immunodeficient mice. The imaging technology uses a modified version of two-step transcriptional amplification (TSTA). The tissue specificity of gene expression is imparted by an enhanced version of the prostate-specific antigen regulatory region that expresses GAL4-ELK1. GAL4-ELK1 confers MAPK specificity by activating a firefly luciferase (FLuc) reporter gene when the Ets-like transcription factor (ELK) 1 activation domain is phosphorylated by MAPK. FLuc activity in live animals was detected using the Xenogen In vivo Imaging System. We validated the TSTA-ELK1 system by analyzing its response to epidermal growth factor treatment in transfected tissue culture cells and in adenovirus (AdTSTA-ELK1)-injected prostate cancer xenograft tumors. We measured MAPK activity in two well characterized xenograft models, CWR22 and LAPC9. Although no significant differences in MAPK levels were detected between androgen-dependent and androgen independent xenografts, the CWR22 models display significantly higher levels of AdTSTA-ELK1 activity versus LAPC9. Western blots of tumor extracts showed that the elevated imaging signal in CWR22 xenografts correlated with elevated levels of phosphorylated extracellular signal-regulated kinase 1/2 but not p38 or c-Jun NH(2)-terminal kinase. We conclude that a gene expression-based optical imaging system can accurately detect and quantify MAPK activity in live animals. PMID- 17108115 TI - A novel SV40 TAg transgenic model of asbestos-induced mesothelioma: malignant transformation is dose dependent. AB - Although it has been clear for >40 years that mesothelioma can be caused by asbestos, not all patients with this disease have a history of asbestos exposure. Other factors, including non-asbestos fibers and ionizing radiation, are known to cause malignant transformation of mesothelial cells. In addition, it is likely that genetics will play some role in susceptibility. Recently, it has been suggested that SV40 viral oncogenes could contribute to the carcinogenicity of asbestos. To better understand the role of SV40, we used the mesothelin promoter to construct MexTAg mice that express SV40 large T antigen (TAg) in the mesothelial compartment. We generated four MexTAg lines that carry high, intermediate, and low copy numbers of the transgene. All of these mice show a relatively low level of spontaneous tumor development. High-copy, 299h mice rapidly developed mesotheliomas when exposed to asbestos, and these tumors were faster growing and more invasive than those developing in wild-type and single copy (266s) mice. In addition, we found a direct relationship between transgene copy number and survival after exposure to asbestos. A single copy of TAg was sufficient to immortalize mesothelial cells in vitro, but these cells did not show evidence of malignant transformation. In contrast, cell lines developed from mesothelial cells of animals carrying multiple copies of TAg were growth factor independent and could be cloned at limiting dilution in soft agar. These data provide the first in vivo demonstration of co-carcinogenicity between SV40 and asbestos. PMID- 17108116 TI - Mass spectrometry-based metabolic profiling reveals different metabolite patterns in invasive ovarian carcinomas and ovarian borderline tumors. AB - Metabolites are the end products of cellular regulatory processes, and their levels can be regarded as the ultimate response of biological systems to genetic or environmental changes. We have used a metabolite profiling approach to test the hypothesis that quantitative signatures of primary metabolites can be used to characterize molecular changes in ovarian tumor tissues. Sixty-six invasive ovarian carcinomas and nine borderline tumors of the ovary were analyzed by gas chromatography/time-of-flight mass spectrometry (GC-TOF MS) using a novel contamination-free injector system. After automated mass spectral deconvolution, 291 metabolites were detected, of which 114 (39.1%) were annotated as known compounds. By t test statistics with P < 0.01, 51 metabolites were significantly different between borderline tumors and carcinomas, with a false discovery rate of 7.8%, estimated with repeated permutation analysis. Principal component analysis (PCA) revealed four principal components that were significantly different between both groups, with the highest significance found for the second component (P = 0.00000009). PCA as well as additional supervised predictive models allowed a separation of 88% of the borderline tumors from the carcinomas. Our study shows for the first time that large-scale metabolic profiling using GC TOF MS is suitable for analysis of fresh frozen human tumor samples, and that there is a consistent and significant change in primary metabolism of ovarian tumors, which can be detected using multivariate statistical approaches. We conclude that metabolomics is a promising high-throughput, automated approach in addition to functional genomics and proteomics for analyses of molecular changes in malignant tumors. PMID- 17108117 TI - Oxidative stress induces premature senescence by stimulating caveolin-1 gene transcription through p38 mitogen-activated protein kinase/Sp1-mediated activation of two GC-rich promoter elements. AB - Cellular senescence is believed to represent a natural tumor suppressor mechanism. We have previously shown that up-regulation of caveolin-1 was required for oxidative stress-induced premature senescence in fibroblasts. However, the molecular mechanisms underlying caveolin-1 up-regulation in senescent cells remain unknown. Here, we show that subcytotoxic oxidative stress generated by hydrogen peroxide application promotes premature senescence and stimulates the activity of a (-1,296) caveolin-1 promoter reporter gene construct in fibroblasts. Functional deletion analysis mapped the oxidative stress response elements of the mouse caveolin-1 promoter to the sequences -244/-222 and -124/ 101. The hydrogen peroxide-mediated activation of both Cav-1 (-244/-222) and Cav 1 (-124/-101) was prevented by the antioxidant quercetin. Combination of electrophoretic mobility shift studies, chromatin immunoprecipitation analysis, Sp1 overexpression experiments, as well as promoter mutagenesis identifies enhanced Sp1 binding to two GC-boxes at -238/-231 and -118/-106 as the core mechanism of oxidative stress-triggered caveolin-1 transactivation. In addition, signaling studies show p38 mitogen-activated protein kinase (MAPK) as the upstream regulator of Sp1-mediated activation of the caveolin-1 promoter following oxidative stress. Inhibition of p38 MAPK prevents the oxidant-induced Sp1-mediated up-regulation of caveolin-1 protein expression and development of premature senescence. Finally, we show that oxidative stress induces p38-mediated up-regulation of caveolin-1 and premature senescence in normal human mammary epithelial cells but not in MCF-7 breast cancer cells, which do not express caveolin-1 and undergo apoptosis. This study delineates for the first time the molecular mechanisms that modulate caveolin-1 gene transcription upon oxidative stress and brings new insights into the redox control of cellular senescence in both normal and cancer cells. PMID- 17108118 TI - Oncogenic serine-threonine kinase receptor-associated protein modulates the function of Ewing sarcoma protein through a novel mechanism. AB - Although much is known about the oncogenic functions of chimeric Ewing sarcoma (EWS) fusion proteins that result from chromosomal translocations, the cellular role of the normal EWS protein is not well characterized. We have previously identified a WD domain-containing protein, serine-threonine kinase receptor associated protein (STRAP), which inhibits transforming growth factor beta (TGF beta) signaling through interaction with receptors and Smad7 and promotes growth and enhances tumorigenicity. Here, we report the interaction between STRAP and EWS using matrix-assisted laser desorption/ionization, time-of-flight and tandem mass spectrometry. Although STRAP is localized in both cytoplasm and nucleus, nuclear STRAP colocalizes and associates specifically with EWS in the nucleus through its NH(2) and COOH termini. We have found that normal EWS protein is up regulated in human cancers, which correlates with the up-regulation of STRAP in 71% of colorectal cancers and 54% of lung cancers, suggesting a cooperative role of these two proteins in human cancers. TGF-beta has no effect on STRAP and EWS interaction. However, EWS, like STRAP, attenuates TGF-beta-dependent transcription. STRAP inhibits EWS-dependent p300-mediated transactivation of EWS target genes, such as ApoCIII and c-fos, in a TGF-beta-independent manner. Interestingly, we have shown that STRAP blocks the interaction between EWS and p300, whereas the complex formation between STRAP and EWS is not affected by p300. These results suggest that STRAP inhibits the transactivation function of EWS by displacing p300 from the functional transcriptional complex. Thus, this study provides a novel TGF-beta-independent function of STRAP and describes a mechanism by which STRAP regulates the function of oncogenic EWS protein. PMID- 17108119 TI - Cyclooxygenase-2 inhibition suppresses alphavbeta6 integrin-dependent oral squamous carcinoma invasion. AB - Worldwide oral squamous cell carcinoma (OSCC) represents about 5.5% of all malignancies, with approximately 30,000 new cases each year in the United States. The integrin alpha(v)beta(6) and the enzyme cyclooxygenase-2 (COX-2) are implicated in OSCC progression and have been suggested as possible therapeutic targets. Each protein also is reported to identify dysplasias at high risk of malignant transformation, and current clinical trials are testing the efficacy of nonsteroidal anti-inflammatory drugs (NSAID) at preventing OSCC development. Given the probable increased expression of alpha(v)beta(6) and COX-2 in OSCC and the inhibition of several integrins by NSAIDs, we investigated whether NSAIDs affected alpha(v)beta(6)-dependent cell functions. We found that expression of both alpha(v)beta(6) and COX-2 was significantly higher in OSCC compared with oral epithelial dysplasias. Neither protein preferentially identified those dysplastic lesions that became malignant. Using OSCC cell lines, modified to express varying levels of alpha(v)beta(6), we assessed the effect of COX-2 inhibition on cell invasion. We found that the COX-2 inhibitor NS398 inhibited specifically alpha(v)beta(6)-dependent, but not alpha(v)beta(6)-independent, OSCC invasion in vitro and in vivo, and this effect was modulated through prostaglandin E(2) (PGE(2))-dependent activation of Rac-1. Transient expression of constitutively active Rac-1, or addition of the COX-2 metabolite PGE(2), prevented the anti-invasive effect of NS398. Conversely, RNA interference down regulation of Rac-1 inhibited alpha(v)beta(6)-dependent invasion. These findings suggest that COX-2 and alpha(v)beta(6) interact in promoting OSCC invasion. This is a novel mechanism that, given the ubiquity of alpha(v)beta(6) expression by head and neck cancers, raises the possibility that NSAIDs could protect against OSCC invasion. PMID- 17108120 TI - MicroRNA responses to cellular stress. AB - Recent work has begun to explore the instrumental role that small noncoding RNA species, particularly microRNAs (miRNA), have both in classifying human tumors and in directing embryonic development. These studies suggest that developmental programs in essentially all organisms studied are set, in part, by varied expressions of miRNAs and that neoplasia is characterized by altered expression of miRNAs. Reasoning that these observations are linked, we examined whether cellular exposures that induce both developmental anomalies and cancer alter miRNAs. Using microarrays of 385 known human miRNAs, we studied human lymphoblastoid cells grown under various conditions or treatments. Folate deficiency induced a pronounced global increase in miRNA expression. We observed no significant alteration in miRNA expression in cells treated with gamma irradiation, whereas exposure to sodium arsenite led to global increases in miRNA expression. The miRNA hsa-miR-222 was identified from these arrays as significantly overexpressed under folate-deficient conditions, and this finding was confirmed in vivo in human peripheral blood from individuals with low folate intake. Alterations to cellular miRNA expression profiles represent a novel mode of action of folate deprivation and arsenic exposure, and specific alterations in miRNA expression may be a powerful biomarker for these and other toxins with serious effects on human health. PMID- 17108121 TI - Spontaneous transformation of cultured mouse bone marrow-derived stromal cells. AB - Bone marrow-derived stromal cells have engendered interest because of their therapeutic potential for promoting tissue vascularization and repair. When mononuclear cells isolated from mouse bone marrow were cultured in DMEM supplemented with 10% fetal bovine serum, cell populations arose that showed rapid proliferation and loss of contact inhibition. These cells formed invasive soft tissue sarcomas after i.m. injection into nude or scid mice. I.v. injection resulted in the formation of tumor foci in the lungs. The tumors were transplantable into syngeneic immunocompetent mice. Direct injection of cultured cells into immunocompetent mice also resulted in tumor formation. Karyotype analysis showed that increased chromosome number and multiple Robertsonian translocations occurred at passage 3 coincident with the loss of contact inhibition. The remarkably rapid malignant transformation of cultured mouse bone marrow cells may have important implications for ongoing clinical trials of cell therapy and for models of oncogenesis. PMID- 17108122 TI - Metabolic imaging by hyperpolarized 13C magnetic resonance imaging for in vivo tumor diagnosis. AB - The "Warburg effect," an elevation in aerobic glycolysis, may be a fundamental property of cancer cells. For cancer diagnosis and treatment, it would be valuable if elevated glycolytic metabolism could be quantified in an image in animals and humans. The pyruvate molecule is at the metabolic crossroad for energy delivery inside the cell, and with a noninvasive measurement of the relative transformation of pyruvate into lactate and alanine within a biologically relevant time frame (seconds), it may be possible to quantify the glycolytic status of the cells. We have examined the metabolism after i.v. injection of hyperpolarized (13)C-pyruvate in rats with implanted P22 tumors. The strongly enhanced nuclear magnetic resonance signal generated by the hyperpolarization techniques allows mapping of pyruvate, lactate, and alanine in a 5 x 5 x 10 mm(3) imaging voxel using a 1.5 T magnetic resonance scanner. The magnetic resonance scanning (chemical shift imaging) was initiated 24 seconds after the pyruvate injection and had a duration of 14 seconds. All implanted tumors showed significantly higher lactate content than the normal tissue. The results indicate that noninvasive quantification of localized Warburg effect may be possible. PMID- 17108123 TI - Reassessment of id1 protein expression in human mammary, prostate, and bladder cancers using a monospecific rabbit monoclonal anti-id1 antibody. AB - Id proteins are a class of dominant-negative antagonists of helix-loop-helix transcription factors and have been shown to control differentiation of a variety of cell types in diverse organisms. Although the importance of Id1 in tumor endothelial cells is well established, the expression and role of the Id1 protein in human cancer cells is controversial. To explore this issue, we developed and characterized a highly specific rabbit monoclonal antibody against Id1 to assess its expression in human breast, prostate, and bladder malignancies. Our results show that in usual types of human mammary carcinomas, the Id1 protein is expressed exclusively in the endothelium. Interestingly, we detected nuclear expression of the Id1 protein in the tumor cells in 10 of 45 cases of poorly differentiated and highly aggressive carcinoma with metaplastic morphology. Similarly, only 1 of 30 prostate cancer samples showed Id1-positive tumor cells, whereas in almost all, endothelial cells showed high Id1 expression. Intriguingly, whereas normal prostate glands do not show any Id1 protein expression, basal layer cells of benign prostate glands in proximity to tumors expressed high levels of the Id1 protein. In contrast to the lack of Id1 expression in the usual types of mammary and prostate cancers, the majority of transitional cell bladder tumors showed Id1 protein expression in both tumor and endothelial cells. These results suggest that further refinement of Id1 expression patterns in a variety of tumor types will be necessary to identify and study the functional roles played by Id1 in human neoplastic processes. PMID- 17108124 TI - Phosphorylation by DNA-dependent protein kinase is critical for apoptosis induction by insulin-like growth factor binding protein-3. AB - Insulin-like growth factor (IGF) binding protein-3 (IGFBP-3) promotes apoptosis of cancer cells by both IGF-dependent and IGF-independent mechanisms. In vitro phosphorylation of IGFBP-3 by DNA-dependent protein kinase (DNA-PK) has been reported but with unknown functional relevance. Using a chemical inhibitor for DNA-PK in prostate cancer cells and a paired system of glioblastoma cell lines that either lack or express DNA-PK, we show that the apoptosis-promoting and growth-inhibitory actions of IGFBP-3 are completely abrogated in the absence of catalytically active DNA-PK. In the absence of DNA-PK activity, IGFBP-3 has reduced nuclear accumulation and is unable to bind its nuclear binding partner retinoid X receptor (RXR) alpha. We assessed the importance of the three potential DNA-PK phosphorylation sites in IGFBP-3 using PCR-based site-directed mutagenesis. When transfected into 22RV1 cells, IGFBP-3-S165A and IGFBP-3-T170A functioned in an identical manner to wild-type IGFBP-3 to induce apoptosis. In contrast, IGFBP-3-S156A was unable to promote apoptosis and exhibited reduced nuclear accumulation, suggesting a key role for DNA-PK-dependent phosphorylation in the regulation of IGFBP-3 action. These studies reveal a novel regulatory mechanism for the actions of IGFBP-3 in prostate cancer and show phosphorylation of Ser(156) to be functionally critical in its apoptosis-inducing actions. PMID- 17108125 TI - In vivo activity of the cleaved form of soluble urokinase receptor: a new hematopoietic stem/progenitor cell mobilizer. AB - Cleaved forms of soluble urokinase receptor (c-suPAR) have been detected in body fluids from patients affected by various tumors. We recently reported increased c suPAR levels in sera of healthy donors during granulocyte colony-stimulating factor (G-CSF)-induced mobilization of CD34(+) hematopoietic stem cells (HSC). In vitro, c-suPAR or its derived chemotactic peptide (uPAR(84-95)) stimulated migration of human CD34(+) HSCs and inactivated CXCR4, the chemokine receptor primarily responsible for HSC retention in bone marrow. These results suggested that c-suPAR could potentially contribute to regulate HSC trafficking from and to bone marrow. Therefore, we investigated uPAR(84-95) effects on mobilization of mouse CD34(+) hematopoietic stem/progenitor cells (HSC/HPC). We first showed that uPAR(84-95) stimulated in vitro dose-dependent migration of mouse CD34(+) M1 leukemia cells and inactivated murine CXCR4. uPAR(84-95) capability to induce mouse HSC/HPC release from bone marrow and migration into the circulation was then investigated in vivo. uPAR(84-95) i.p. administration induced rapid leukocytosis, which was associated with an increase in peripheral blood CD34(+) HSCs/HPCs. In vitro colony assays confirmed that uPAR(84-95) mobilized hematopoietic progenitors, showing an absolute increase in circulating colony forming cells. uPAR(84-95) mobilizing activity was comparable to that of G-CSF; however, neither synergistic nor additive effect was observed in combining the two molecules. These findings show for the first time in vivo biological effects of c-suPAR. Its capability to mobilize HSCs suggests potential clinical applications in HSC transplantation. PMID- 17108126 TI - Oncostatin M (OSM) cytostasis of breast tumor cells: characterization of an OSM receptor beta-specific kernel. AB - The interleukin-6 cytokine oncostatin M (OSM) induces potent growth-inhibitory and morphogenic responses in several different tumor cell types, highlighting the importance of OSM signaling mechanisms as targets for therapeutic intervention. The specific molecular pathways involved are not well understood, as OSM can signal through two separate heterodimeric receptor complexes, glycoprotein 130 (gp130)/leukemia inhibitory factor receptor (LIFR) alpha and gp130/OSM receptor beta (OSMRbeta). In this investigation, we used a LIFR antagonist to help resolve signaling responses and identify patterns of gene expression elicited by the different receptor complexes. OSM-induced biological effects on breast tumor derived cell lines were specifically mediated through the gp130/OSMRbeta complex. Each cytokine tested exhibited differential signaling capability and manifested both shared and unique patterns of gene activation, emphasizing compositional differences in activator protein-1 transcription factor activity and expression. In particular, OSM strongly activated the c-Jun NH(2)-terminal kinase (JNK) serine/threonine kinase and downstream components, including activating transcription factor (ATF)/cyclic AMP-responsive element binding protein family member, ATF3. JNK/stress-activated protein kinase kinase inhibition abrogated cell morphogenesis induced by OSM, indicating an important role for this pathway in OSM specificity. These findings identify a core signaling/transcriptional mechanism specific to the OSMRbeta in breast tumor cells. PMID- 17108127 TI - Gonadotropin-releasing hormone promotes ovarian cancer cell invasiveness through c-Jun NH2-terminal kinase-mediated activation of matrix metalloproteinase (MMP)-2 and MMP-9. AB - Gonadotropin-releasing hormone (GnRH) receptor is present in 80% of ovarian cancer, and numerous studies have provided evidence for a role of GnRH in cell proliferation. In this study, the effect of GnRH on the invasion potential of ovarian cancer cells was investigated. In vitro migration and cell invasion assays with the ovarian cancer cell lines Caov-3 and OVCAR-3 revealed the biphasic nature of GnRH; low concentrations of GnRH agonist (GnRHa) increased the cell motility and invasiveness of these cells, but at increased concentrations, the stimulatory effect was insignificant. Reverse transcription-PCR, Western blot, and gelatin zymography showed that the expression of metastasis-related proteinases, matrix metalloproteinase (MMP)-2 and MMP-9, was up-regulated and activated by GnRHa. Moreover, we observed that GnRHa was able to transactivate the MMP-2 and MMP-9 promoters. The invasive/migratory phenotype activated by GnRHa can be blocked by specific inhibitors or neutralizing antibodies to MMP-2 and MMP-9. Knockdown of the GnRH receptor using small interfering RNA significantly inhibited the GnRH-induced MMP activation, invasion, and migration. In addition, we showed that the c-Jun NH(2)-terminal kinase, but not extracellular signal-regulated kinase 1/2 or p38 mitogen-activated protein kinase, signaling pathway was critical for GnRH-mediated up-regulation of MMP, cell invasion, and motility. These results indicate for the first time an expanded role for GnRH in other aspects of ovarian tumor progression, such as metastasis, via activation of MMP and the subsequent increase in cell migration and invasion. PMID- 17108128 TI - Cytosine methylation profiles as a molecular marker in non-small cell lung cancer. AB - Aberrant promoter methylation is frequently observed in different types of lung cancer. Epigenetic modifications are believed to occur before the clinical onset of the disease and hence hold a great promise as early detection markers. Extensive analysis of DNA methylation has been impeded by methods that are either too labor intensive to allow large-scale studies or not sufficiently quantitative to measure subtle changes in the degree of methylation. We used a novel quantitative DNA methylation analysis technology to complete a large-scale cytosine methylation profiling study involving 47 gene promoter regions in 96 lung cancer patients. Each individual contributed a lung cancer specimen and corresponding adjacent normal tissue. The study identified six genes with statistically significant differences in methylation between normal and tumor tissue (P < 10(-6)). We explored the quantitative methylation data using an unsupervised hierarchical clustering algorithm. The data analysis revealed that methylation patterns differentiate normal from tumor tissue. For validation of our approach, we divided the samples to train a classifier and test its performance. We were able to distinguish normal from lung cancer tissue with >95% sensitivity and specificity. These results show that quantitative cytosine methylation profiling can be used to identify molecular classification markers in lung cancer. PMID- 17108129 TI - Inhibition of prostate tumor growth and bone remodeling by the vascular targeting agent VEGF121/rGel. AB - The pathophysiology of tumor growth following skeletal metastases and the poor response of this type of lesion to therapeutic intervention remains incompletely understood. Vascular endothelial growth factor (VEGF)-A and its receptors play a role in both osteoclastogenesis and tumor growth. Systemic (i.v.) treatment of nude mice bearing intrafemoral prostate (PC-3) tumors with the vascular ablative agent VEGF(121)/recombinant gelonin (rGel) strongly inhibited tumor growth. Fifty percent of treated animals had complete regression of bone tumors with no development of lytic bone lesions. Immunohistochemical analysis showed that VEGF(121)/rGel treatment suppressed tumor-mediated osteoclastogenesis in vivo. In vitro treatment of murine osteoclast precursors, both cell line (RAW264.7) and bone marrow-derived monocytes (BMM), revealed that VEGF(121)/rGel was selectively cytotoxic to osteoclast precursor cells rather than mature osteoclasts. VEGF(121)/rGel cytotoxicity was mediated by Flt-1, which was down-regulated during osteoclast differentiation. Analysis by flow cytometry and reverse transcription-PCR showed that both BMM and RAW264.7 cells display high levels of Flt-1 but low levels of Flk-1. Internalization of VEGF(121)/rGel into osteoclast precursor cells was suppressed by pretreatment with an Flt-1 neutralizing antibody or by placenta growth factor but not with an Flk-1 neutralizing antibody. Thus, VEGF(121)/rGel inhibits osteoclast maturation in vivo and it seems that this process is important in the resulting suppression of skeletal osteolytic lesions. This is a novel and unique mechanism of action for this class of agents and suggests a potentially new approach for treatment or prevention of tumor growth in bone. PMID- 17108130 TI - Osteoclasts direct bystander killing of bone cancer. AB - Primary and metastatic bone cancers are difficult to eradicate and novel approaches are needed to improve treatment and extend life. As bone cancer grows, osteoclasts, the principal bone-resorbing cells of the body, are recruited to and activated at sites of cancer. In this investigation, we determined if osteoclast lineage cells could function as a cell-based gene delivery system to bone cancers. We used the cytosine deaminase (CD) 5-fluorocytosine (5-FC) enzyme/prodrug system and studied bone marrow and bones from transgenic mice expressing a novel CD gene regulated by the osteoclast tartrate-resistant acid phosphatase (TRAP) gene promoter (Tg/NCD). DsRed2-labeled 2472 sarcoma cells were placed in Tg/NCD osteoclastogenic cultures and treated with 5-FC. 5-FC treatment resulted in profound bystander killing (90%; P < 0.05). The effect of 5-FC treatment on osteoclast lineage cells was most dramatic when administered at the beginning of the 7-day cultures, suggesting that mature osteoclasts are less sensitive to 5-FC. Evaluation of osteoclast-directed bystander killing in vivo revealed dramatic killing of bone cancer with only a modest effect on osteoclast number. Specifically, 5-FC treatment of tumor-bearing Tg/NCD mice or Tg/NCD bone marrow transplanted C3H mice (Tg/NCD-C3H) resulted in 92% and 44% reductions in tumor area, respectively (P < 0.05). Eight of ten 5-FC-treated Tg/NCD mice had complete bone tumor killing and five of six 5-FC-treated Tg/NCD-C3H mice had reduced tumor compared with controls. In addition, Tg/NCD osteoclasts were resistant to 5-FC treatment in vivo, a very important feature, as it identifies osteoclasts as an ideal CD gene delivery system. PMID- 17108131 TI - Reversal of drug resistance in breast cancer cells by transglutaminase 2 inhibition and nuclear factor-kappaB inactivation. AB - Induction of transglutaminase 2 (TGase 2) by epidermal growth factor (EGF) in human breast cancer cells increases their oncogenic potential and chemoresistance. The role of TGase 2 in the development of these tumor-related phenotypes remains to be elucidated, but it has been shown that expression of a dominant-negative form of TGase 2 reverses EGF-mediated chemoresistance in breast cancer cells. We examined several different breast cancer cell lines, representing both EGF receptor (EGFR)-positive and EGFR-negative breast cancers, and found that doxorubicin-resistant cells had a higher level of TGase 2 compared with doxorubicin-sensitive cells independent of the EGFR expression level. TGase 2 inhibition increased the chemosensitivity of drug-resistant cells, concomitant with a decrease in nuclear factor-kappaB (NF-kappaB) activity. Increasing the level of TGase 2 in drug-sensitive cells by transient transfection reduced the level of inhibitory subunit alpha of NF-kappaB (IkappaBalpha) and increased NF kappaB activity in these cells. Inhibition of TGase 2 in drug-resistant cells by RNA interference increased the levels of IkappaBalpha, and this correlated with a shift in the accumulation of NF-kappaB from the nucleus to the cytosol. We recently showed that TGase 2 activated NF-kappaB through polymerization and depletion of free IkappaBalpha during inflammation. Therefore, increased expression of TGase 2 and subsequent activation of NF-kappaB may contribute to drug resistance in breast cancer cells independently of EGF signaling. PMID- 17108132 TI - The internalization and degradation of human copper transporter 1 following cisplatin exposure. AB - The human copper transporter 1 (hCTR1), the major transporter responsible for copper influx, mediates one component of the cellular accumulation of cisplatin (DDP). Both copper and DDP cause rapid down-regulation of hCTR1 expression in human ovarian carcinoma cells. In this study, we investigated the mechanism of this effect using digital deconvolution microscopy and Western blot analysis of cells stained with antibodies directed at both ends of the protein. Treatment of 2008 cells with DDP in combination with inhibitors of various endosomal pathways (amiloride, cytochalasin D, nystatin, and methyl-beta-cyclodextrin) showed that hCTR1 degradation was blocked by amiloride and cytochalasin D, indicating that hCTR1 was internalized primarily by macropinocytosis. Expression of transdominant negative forms of dynamin I and Rac showed that loss of hCTR1 was not dependent on pathways regulated by either of these proteins. DDP-induced loss of hCTR1 was blocked by the proteasome inhibitors lactacystin, proteasome inhibitor 1, and MG132. This study confirms that DDP triggers the rapid loss of hCTR1 from ovarian carcinoma cells at clinically relevant concentrations. The results indicate that DDP-induced loss of hCTR1 involves internalization from the plasma membrane by macropinocytosis followed by proteasomal degradation. Because hCTR1 is a major determinant of early DDP uptake, prevention of its degradation offers a potential approach to enhancing tumor sensitivity. PMID- 17108133 TI - A mechanism-based combination therapy reduces local tumor growth and metastasis in an orthotopic model of prostate cancer. AB - Therapy-induced stimulation of angiogenic molecules can promote tumor angiogenesis leading to enhanced tumor growth and cancer metastasis. Several standard and emerging therapies, such as radiation and photodynamic therapy (PDT), can induce angiogenic molecules, thus limiting their effectiveness. PDT is approved for the treatment of several cancers; however, its induction of vascular endothelial growth factor (VEGF) creates conditions favorable to enhanced tumor growth and metastasis, therefore mitigating its cytotoxic and antivascular effects. This is the first report showing that subcurative PDT in an orthotopic model of prostate cancer (LNCaP) increases not only VEGF secretion (2.1-fold) but also the fraction of animals with lymph node metastases. PDT followed by administration of an antiangiogenic agent, TNP-470, abolished this increase and reduced local tumor growth. On the other hand, administration of TNP-470 before PDT was less effective at local tumor control. In addition, animals in all groups, except in the PDT + TNP-470 group, had a weight loss of >3 g at the time of sacrifice; the weight of the animals in the PDT + TNP-470 group did not change. The significant reduction (P < 0.05) in tumor weight and volume observed between the PDT + TNP-470 group and the control group suggests that the combination of PDT and antiangiogenic treatment administered in the appropriate sequence was not only more effective at controlling local tumor growth and metastases but also reduced disease-related toxicities. Such molecular response based combinations merit further investigations as they enhance both monotherapies and lead to improved treatment outcomes. PMID- 17108134 TI - A sequential blockade strategy for the design of combination therapies to overcome oncogene addiction in chronic myelogenous leukemia. AB - Some tumors are dependent on the continued activity of a single oncogene for maintenance of their malignant phenotype. The best-studied example is the Bcr-Abl fusion protein in chronic myelogenous leukemia (CML). Although the clinical success of the Abl kinase inhibitor imatinib against chronic-phase CML emphasizes the importance of developing therapeutic strategies aimed at this target, resistance to imatinib poses a major problem for the ultimate success of CML therapy by this agent. We hypothesized a sequential blockade strategy that is designed to decrease the expression of the Bcr-Abl protein, with the goal of complementing the action of imatinib on kinase activity. In this study, flavopiridol, an inhibitor of transcription, homoharringtonine (HHT), a protein synthesis inhibitor, and imatinib were used singly and in combination against the Bcr-Abl-positive human CML cell line K562. Flavopiridol alone inhibited phosphorylation of the RNA polymerase II COOH-terminal domain, specifically reduced RNA polymerase II-directed mRNA synthesis, and decreased the Bcr-Abl transcript levels. HHT inhibited protein synthesis and reduced the Bcr-Abl protein level. Imatinib directly inhibited the kinase activity of Bcr-Abl. The combinations of flavopiridol and HHT and flavopiridol and imatinib synergistically decreased clonogenicity as evaluated by the median-effect method. Greater synergy was observed when HHT and imatinib were given sequentially compared with simultaneous administration. Imatinib-resistant Ba/F3 cells that were transfected to express the E255K and T315I mutations of Bcr-Abl were not cross-resistant to flavopiridol and HHT. These results provided a rationale for the combination of inhibitors of transcription and/or translation with specific kinase inhibitors. PMID- 17108135 TI - Up-regulation of heat shock protein 27 induces resistance to 17-allylamino demethoxygeldanamycin through a glutathione-mediated mechanism. AB - 17-Allylamino-demethoxygeldanamycin (17-AAG), currently in phase I and II clinical trials as an anticancer agent, binds to the ATP pocket of heat shock protein (Hsp90). This binding induces a cellular stress response that up regulates many proteins including Hsp27, a member of the small heat shock protein family that has cytoprotective roles, including chaperoning of cellular proteins, regulation of apoptotic signaling, and modulation of oxidative stress. Therefore, we hypothesized that Hsp27 expression may affect cancer cell sensitivity to 17 AAG. In colony-forming assays, overexpression of Hsp27 increased cell resistance to 17-AAG whereas down-regulation of Hsp27 by siRNA increased sensitivity. Because Hsp27 is known to modulate levels of glutathione (GSH), we examined cellular levels of GSH and found that it was decreased in cells transfected with Hsp27 siRNA when compared with control siRNA. Treatment with buthionine sulfoximine, an inhibitor of GSH synthesis, also sensitized cells to 17-AAG. Conversely, treatment of Hsp27 siRNA-transfected cells with N-acetylcysteine, an antioxidant and GSH precursor, reversed their sensitivity to 17-AAG. A cell line selected for stable resistance to geldanamycin relative to parent cells showed increased Hsp27 expression. When these geldanamycin- and 17-AAG-resistant cells were transfected with Hsp27 siRNA, 17-AAG resistance was dramatically diminished. Our results suggest that Hsp27 up-regulation has a significant role in 17-AAG resistance, which may be mediated in part through GSH regulation. Clinical modulation of GSH may therefore enhance the efficacy of Hsp90-directed therapy. PMID- 17108136 TI - Biological properties of IDN5174, a new synthetic camptothecin with the open lactone ring. AB - A series of water-soluble camptothecins obtained by linking a spermidine moiety to the 21-position of the open form through an amidic bond have been tested for their biochemical and biological activities. Growth inhibition assay on the human non-small cell lung cancer carcinoma NCI-H460 cell line revealed that the camptothecin analogues were less potent than topotecan and SN38 after 1 hour of treatment. The potency increased after 72 hours of exposure, being similar to that of reference camptothecins. The analysis of topoisomerase I-mediated DNA cleavage using the purified enzyme indicated that the novel camptothecin analogues retained ability to poison topoisomerase I and displayed the same cleavage pattern of SN38. Persistence of the DNA cleavage was comparable with that of SN38. Stabilization of the cleavable complex was not the result of hydrolysis of the N-C bond between polyamine and the drug because no free camptothecin was recovered at the end of DNA cleavage in presence of IDN5174, the analogue selected for detailed studies. IDN5174 exhibited an antitumor activity comparable with that of topotecan and irinotecan against NCI-H460 tumor xenograft. The pharmacokinetics in mice showed a favorable disposition in tumor tissue with low amount of camptothecin detectable in plasma and tumor (around 5 10%), thus supporting the efficacy of intact IDN5174. In conclusion, we found that IDN5174 maintained the biological and antitumor properties, in spite of lack of the closed E ring. The available results support the interpretation that the polyamine linked at the 21-position may allow a favorable drug interaction in the ternary complex. PMID- 17108137 TI - Generation of a stable antioxidant response element-driven reporter gene cell line and its use to show redox-dependent activation of nrf2 by cancer chemotherapeutic agents. AB - The NF-E2 p45-related factor 2 (Nrf2) regulates cytoprotective genes that contain an antioxidant response element (ARE) in their promoters. To investigate whether anticancer drugs can induce ARE-driven gene expression, we have developed a stable human mammary MCF7-derived reporter cell line called AREc32, which contains a luciferase gene construct controlled by eight copies of the cis element. In these cells, luciferase activity was increased up to 50-fold following treatment with 50 mumol/L tert-butylhydroquinone (t-BHQ). Basal and inducible luciferase activities in AREc32 cells were increased by forced overexpression of Nrf2 and reduced by knockdown of endogenous Nrf2 expression with RNA interference. Depletion of cellular reduced glutathione (GSH) by treatment of AREc32 cells with l-buthionine-S,R-sulfoximine (BSO) did not influence basal levels of luciferase activity, but pretreatment with BSO augmented induction of luciferase activity by t-BHQ. Induction of reporter activity by t-BHQ in AREc32 cells was suppressed markedly by the antioxidants N acetylcysteine and GSH but only modestly by vitamins C or E, suggesting that ARE luciferase expression is induced primarily by thiol-active electrophiles rather than free radicals. The anticancer drugs cisplatin, etoposide, mitoxantrone, chlorambucil, melphalan, and carmustine [1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU)] weakly induced luciferase activity in AREc32 cells. Moreover, treatment of AREc32 cells with BSO immediately before exposure to anticancer drugs enhanced induction of ARE-driven luciferase activity by cisplatin, BCNU, chlorambucil, and melphalan and also induced endogenous AKR1C (AKR1C refers to AKR1C1 and AKR1C2), a target gene of Nrf2. Our findings show that Nrf2 can be activated by certain anticancer agents, and this will influence the effectiveness of chemotherapy. PMID- 17108138 TI - CD28 costimulation provided through a CD19-specific chimeric antigen receptor enhances in vivo persistence and antitumor efficacy of adoptively transferred T cells. AB - Chimeric antigen receptors (CAR) combine an antigen-binding domain with a CD3 zeta signaling motif to redirect T-cell specificity to clinically important targets. First-generation CAR, such as the CD19-specific CAR (designated CD19R), may fail to fully engage genetically modified T cells because activation is initiated by antigen-dependent signaling through chimeric CD3-zeta, independent of costimulation through accessory molecules. We show that enforced expression of the full-length costimulatory molecule CD28 in CD8(+)CD19R(+)CD28(-) T cells can restore fully competent antigen-dependent T-cell activation upon binding CD19(+) targets expressing CD80/CD86. Thus, to provide costimulation to T cells through a CD19-specific CAR, independent of binding to CD80/CD86, we developed a second generation CAR (designated CD19RCD28), which includes a modified chimeric CD28 signaling domain fused to chimeric CD3-zeta. CD19R(+) and CD19RCD28(+) CD8(+) T cells specifically lyse CD19(+) tumor cells. However, the CD19RCD28(+) CD8(+) T cells proliferate in absence of exogenous recombinant human interleukin-2, produce interleukin-2, propagate, and up-regulate antiapoptotic Bcl-X(L) after stimulation by CD19(+) tumor cells. For the first time, we show in vivo that adoptively transferred CD19RCD28(+) T cells show an improved persistence and antitumor effect compared with CD19R(+) T cells. These data imply that modifications to the CAR can result in improved therapeutic potential of CD19 specific T cells expressing this second-generation CAR. PMID- 17108139 TI - Enhanced antitumor response by divergent modulation of natural killer and natural killer T cells in the liver. AB - The use of interleukin-18 (IL-18) together with IL-12 induced high levels of IFN gamma in tumor-bearing mice and regression of liver tumors that was abolished in IFN-gamma((-/-)) mice. Natural killer (NK) and NKT cells were the major producers of IFN-gamma in the livers of mice treated with IL-18 and/or IL-12. Liver NK cells were significantly increased by treatment with IL-18/IL-12, whereas the degree of liver NKT cell TCR detection was diminished by this treatment. Reduction of NK cells with anti-asGM1 decreased the antitumor activity of IL 18/IL-12 therapy and revealed NK cells to be an important component for tumor regression in the liver. In contrast, the antitumor effects of both IL-18 and IL 12 were further increased in CD1d((-/-)) mice, which lack NKT cells. Our data, therefore, show that the antitumor activity induced in mice by IL-18/IL-12 is NK and IFN-gamma dependent and is able to overcome an endogenous immunosuppressive effect of NKT cells in the liver microenvironment. These results suggest that immunotherapeutic approaches that enhance NK cell function while eliminating or altering NKT cells could be effective in the treatment of cancer in the liver. PMID- 17108140 TI - Functional CXCR4-expressing microparticles and SDF-1 correlate with circulating acute myelogenous leukemia cells. AB - Stromal cell-derived factor-1 (SDF-1/CXCL12) and its receptor CXCR4 are implicated in the pathogenesis and prognosis of acute myelogenous leukemia (AML). Cellular microparticles, submicron vesicles shed from the plasma membrane of various cells, are also associated with human pathology. In the present study, we investigated the putative relationships between the SDF-1/CXCR4 axis and microparticles in AML. We detected CXCR4-expressing microparticles (CXCR4(+) microparticles) in the peripheral blood and bone marrow plasma samples of normal donors and newly diagnosed adult AML patients. In samples from AML patients, levels of CXCR4(+) microparticles and total SDF-1 were elevated compared with normal individuals. The majority of CXCR4(+) microparticles in AML patients were CD45(+), whereas in normal individuals, they were mostly CD41(+). Importantly, we found a strong correlation between the levels of CXCR4(+) microparticle and WBC count in the peripheral blood and bone marrow plasma obtained from the AML patients. Of interest, levels of functional, noncleaved SDF-1 were reduced in these patients compared with normal individuals and also strongly correlated with the WBC count. Furthermore, our data indicate NH(2)-terminal truncation of the CXCR4 molecule in the microparticles of AML patients. However, such microparticles were capable of transferring the CXCR4 molecule to AML-derived HL 60 cells, enhancing their migration to SDF-1 in vitro and increasing their homing to the bone marrow of irradiated NOD/SCID/beta2m(null) mice. The CXCR4 antagonist AMD3100 reduced these effects. Our findings suggest that functional CXCR4(+) microparticles and SDF-1 are involved in the progression of AML. We propose that their levels are potentially valuable as an additional diagnostic AML variable. PMID- 17108141 TI - Ciz1, a Novel DNA-binding coactivator of the estrogen receptor alpha, confers hypersensitivity to estrogen action. AB - The transcriptional activity of the estrogen receptor (ER) is affected by regulatory cofactors, including chromatin-remodeling complexes, coactivators, and corepressors. Coregulators are recruited to target gene promoters through protein protein interactions with ER and function as linker molecules between the DNA, DNA-binding proteins, and DNA-modifying enzymes. We recently showed that Cip interacting zinc finger protein 1 (Ciz1) participates in the regulation of the cell cycle in estrogen-stimulated breast cancer cells. Despite the emerging significance of Ciz1 in the biology of breast cancer cells, regulation of endogenous Ciz1 in hormone-responsive cancer cells remains unknown. To shed light on the role of Ciz1 in breast tumorigenesis, we defined the regulation of Ciz1 by the ER pathway and found that Ciz1 is an estrogen-responsive gene. We also discovered that Ciz1 protein, a DNA-binding factor, coregulates ER by enhancing ER transactivation activity by promoting the recruitment of the ER complex to the target gene chromatin. In addition, we found that Ciz1 overexpression confers estrogen hypersensitivity to breast cancer cells and promotes the growth rate, anchorage independency, and tumorigenic properties of breast cancer cells. These findings revealed the inherent role of Ciz1, a novel DNA binding and ER coactivator, in amplifying estrogenic responses and promoting breast cancer tumorigenesis. PMID- 17108142 TI - An inherent role of integrin-linked kinase-estrogen receptor alpha interaction in cell migration. AB - Integrin-linked kinase (ILK) and estrogen receptor (ER)-alpha modulate cell migration. However, the crosstalk between ERalpha and ILK and the role of ILK in ERalpha-mediated cell migration remain unexplored. Here, we report that ILK participates in ERalpha signaling in breast cancer cells. We found that ILK binds ERalpha in vitro and in vivo through a LXXLL motif in ILK. Estrogen prevented ERalpha-ILK binding, resulting in phosphatidylinositol 3-kinase (PI3K)-dependent increase in ILK kinase activity. Furthermore, the regulation of ERalpha-ILK interaction was dependent on the PI3K pathway. Unexpectedly, transient knockdown or inhibition of ILK caused hyperphosphorylation of ERalpha Ser(118) in an extracellular signal-regulated kinase/mitogen-activated protein kinase pathway dependent manner and an enhanced ERalpha recruitment to the target chromatin and gene expression, a process reversed by overexpression of ILK. Compatible with these interactions, estrogen regulated cell migration via the PI3K/ILK/AKT pathway with stable ILK overexpression hyperactivating cell migration. Thus, status of ILK signaling may be an important modifier of ER signaling in breast cancer cells and this pathway could be exploited for therapeutic intervention in breast cancer cells. PMID- 17108143 TI - Steroid receptor coactivator-3 and activator protein-1 coordinately regulate the transcription of components of the insulin-like growth factor/AKT signaling pathway. AB - Steroid receptor coactivator (SRC)-3, also called amplified in breast cancer 1, is a member of the p160 nuclear receptor coactivator family involved in transcriptional regulation of target genes. SRC-3 is frequently amplified and/or overexpressed in hormone-sensitive and hormone-insensitive tumors. We reported previously that SRC-3 stimulated prostate cell growth in a hormone-independent manner through activation of AKT signaling pathway. However, the underlying mechanism remains undefined. Here, we exploited the mifepristone-induced SRC-3 LNCaP prostate cancer cell line generated in our laboratory to identify SRC-3 regulated genes by oligonucleotide microarray analysis. We found that SRC-3 up regulates the expression of multiple genes in the insulin-like growth factor (IGF)/AKT signaling pathway that are involved in cell proliferation and survival. In contrast, knockdown of SRC-3 in PC3 (androgen receptor negative) prostate cancer cells and MCF-7 breast cancer cells reduces their expression. Similarly, in prostate glands of SRC-3 null mice, expressions of these components in the IGF/AKT signal pathway are also reduced. Chromatin immunoprecipitation assay revealed that SRC-3 was directly recruited to the promoters of these genes, indicating that they are direct targets of SRC-3. Interestingly, we showed that recruitment of SRC-3 to two target promoters, IRS-2 and IGF-I, requires transcription factor activator protein-1 (AP-1). Taken together, our results clearly show that SRC-3 and AP-1 can coordinately regulate the transcription of multiple components in the IGF/AKT pathway to ensure ligand-independent cell proliferation and survival of cancer cells. PMID- 17108144 TI - Receptor for activated C kinase 1 (RACK1) and Src regulate the tyrosine phosphorylation and function of the androgen receptor. AB - The androgen receptor (AR) remains functionally important in the development and progression of prostate cancer even when the disease seems androgen "independent." Because signal transduction by growth factor receptors increases in advanced prostate cancer and is capable of sensitizing the AR to androgen, there is considerable interest in determining the mechanisms by which signaling systems can modulate AR function. We show herein that the adaptor/scaffolding protein receptor for activated C kinase 1 (RACK1), which was previously reported to interact with the AR, modulates the tyrosine phosphorylation of AR and its interaction with the Src tyrosine kinase. We also show that down-regulation of RACK1 by short interfering RNA inhibits growth and stimulates prostate-specific antigen transcription in androgen-treated prostate cancer cells. Our results suggest that RACK1 mediates the cross-talk of AR with additional binding partners, such as Src, and facilitates the tyrosine phosphorylation and transcriptional activity of the AR. PMID- 17108145 TI - Molecular imaging of proliferation in malignant lymphoma. AB - We have determined the ability of positron emission tomography (PET) with the thymidine analogue 3'-deoxy-3'-[(18)F]fluorothymidine (FLT) to detect manifestation sites of malignant lymphoma, to assess proliferative activity, and to differentiate aggressive from indolent tumors. In this prospective study, FLT PET was done additionally to routine staging procedures in 34 patients with malignant lymphoma. Sixty minutes after i.v. injection of approximately 330 MBq FLT, emission and transmission scanning was done. Tracer uptake in lymphoma was evaluated semiquantitatively by calculation of standardized uptake values (SUV) and correlated to tumor grading and proliferation fraction as determined by Ki-67 immunohistochemistry. FLT-PET detected a total of 490 lesions compared with 420 lesions revealed by routine staging. In 11 patients with indolent lymphoma, mean FLT-SUV in biopsied lesions was 2.3 (range, 1.2-4.5). In 21 patients with aggressive lymphoma, a significantly higher FLT uptake was observed (mean FLT SUV, 5.9; range, 3.2-9.2; P < 0.0001) and a cutoff value of SUV = 3 accurately discriminated between indolent and aggressive lymphoma. Linear regression analysis indicated significant correlation of FLT uptake in biopsied lesions and proliferation fraction (r = 0.84; P < 0.0001). In this clinical study, FLT-PET was suitable for imaging malignant lymphoma and noninvasive assessment of tumor grading. Due to specific imaging of proliferation, FLT may be a superior PET tracer for detection of malignant lymphoma in organs with high physiologic fluorodeoxyglucose uptake and early detection of progression to a more aggressive histology or potential transformation. PMID- 17108146 TI - DNA repair and cell cycle control genes and the risk of young-onset lung cancer. AB - Exposure to tobacco smoke and to mutagenic xenobiotics can cause various types of DNA damage in lung cells, which, if not corrected by DNA repair systems, may lead to deregulation of the cell cycle and, ultimately, to cancer. Genetic variation could thus be an important factor in determining susceptibility to tobacco induced lung cancer with genetic susceptibility playing a larger role in young onset cases compared with that in the general population. We have therefore studied 102 single-nucleotide polymorphisms (SNP) in 34 key DNA repair and cell cycle control genes in 299 lung cancer cases diagnosed before the age of 50 years and 317 controls from six countries of Central and Eastern Europe. We have found no association of lung cancer risk with polymorphisms in genes related to cell cycle control, single-strand/double-strand break repair, or base excision repair. Significant associations (P < 0.05) were found with polymorphisms in genes involved in DNA damage sensing (ATM) and, interestingly, in four genes encoding proteins involved in mismatch repair (LIG1, LIG3, MLH1, and MSH6). The strongest associations were observed with heterozygote carriers of LIG1 -7C>T [odds ratio (OR), 1.73; 95% confidence interval (95% CI), 1.13-2.64] and homozygote carriers of LIG3 rs1052536 (OR, 2.05; 95% CI, 1.25-3.38). Consideration of the relatively large number of markers assessed diminishes the significance of these findings; thus, these SNPs should be considered promising candidates for further investigation in other independent populations. PMID- 17108147 TI - Persistent human papillomavirus infection is associated with a generalized decrease in immune responsiveness in older women. AB - The development of cervical cancer and its precursors are linked to persistent infection with oncogenic types of human papillomavirus (HPV). Host immune responses seem to be determinants of risk for this disease. However, little is known about the immunologic determinants of HPV persistence. Here, we examined the association between lymphoproliferative responses to antigens/mitogens and persistent HPV infection in women older than 45 years. Women included in this study were participants in a 10,000-woman population-based cohort study of cervical neoplasia in Costa Rica. Women older than 45 years and HPV DNA positive at a screening visit were selected as cases (n = 283). We selected a comparably sized control group of HPV DNA-negative women, matched to cases on age and time since enrollment (n = 261). At an additional clinical visit, women were cytologically and virologically rescreened, and cervical and blood specimens were collected. Proliferative responses to phytohemagglutinin (PHA), influenza virus (Flu), and HPV16 virus-like particle (VLP) were lower among women with persistent HPV infection [median counts per minute (cpm): 72,849 for PHA, 1,241 for Flu, and 727 for VLP] than for the control group (median cpm: 107,049 for PHA, 2,111 for Flu, and 2,068 for VLP). The decreases were most profound in women with long-term persistence and were only observed for the oldest age group (>/=65 years). Our results indicate that an impairment in host immunologic responses is associated to persistent HPV infection. The fact that effects were evident for all studied stimuli is suggestive of a generalized effect. PMID- 17108148 TI - Germ-line genetic variation in the key androgen-regulating genes androgen receptor, cytochrome P450, and steroid-5-alpha-reductase type 2 is important for prostate cancer development. AB - Prostate cancer risk may be influenced by single genetic variants in the hormone regulating genes androgen receptor (AR), cytochrome P450 (CYP17), and steroid-5 alpha-reductase type 2 (SRD5A2). In this study, we comprehensively investigated polymorphisms in these three loci and their joint effect in a large population based study. We selected 23 haplotype-tagging single-nucleotide polymorphisms (htSNP) that could uniquely describe >95% of the haplotypes (6 in AR, 6 in CYP17, and 11 in SRD5A2). These htSNPs were then genotyped in the Cancer Prostate in Sweden population (2,826 case subjects and 1,705 controls). We observed significant association for several SNPs in the AR gene (P = 0.004-0.02) and CYP17 (P = 0.009-0.05) and one SNP in SRD5A2 (P = 0.02). Carriers of the most common AR haplotype had a significant excess risk to develop prostate cancer [odds ratio (OR), 1.25; 95% confidence interval (95% CI), 1.1-1.5; P = 0.002], yielding an estimated population attributable risk of 16% (95% CI, 0.06-0.25). Combining risk alleles from these genes yielded a 12% risk increase for each additional high-risk allele carried (95% CI, 1.1-1.2; P for trend = 9.2 x 10( 5)), with an overall OR of 1.87 (95% CI, 1.0-3.4) for carriers of all five included risk alleles, an OR of 2.13 (P for trend = 8 x 10(-4)) for advanced disease, and an OR of 4.35 (P for trend = 7 x 10(-5)) for disease onset before age 65 years. Genetic variation in key genes in the androgen pathway is important for development of prostate cancer and may account for a considerable proportion of all prostate cancers. Carriers of five high-risk alleles in the AR, CYP17, and SRD5A2 genes are at approximately 2-fold excess risk to develop prostate cancer. PMID- 17108149 TI - Antiemetic activity of corticosteroids in patients receiving cancer chemotherapy: dosing, efficacy, and tolerability analysis. AB - BACKGROUND: Chemotherapy and radiotherapy remain the standards of treatment for many patients with cancer, but these modalities are often limited by distressing side-effects, most notably chemotherapy-induced nausea and vomiting (CINV). METHODS: This paper considers the role of corticosteroids in CINV prophylaxis. Clinical trial results and treatment guidelines indicate that even with the emergence of new serotonin and neurokinin receptor antagonists, corticosteroids continue to play an important role in antiemesis for oncology patients. Numerous clinical trial results have demonstrated that both dexamethasone and methylprednisolone are effective as monotherapy and in combination with older and more recently developed antiemetic agents in patients receiving a wide range of chemotherapeutic regimens used for treatment of different cancers. CONCLUSIONS: With the increasing number of antineoplastic regimens and factors specific to individual patients, it is important to frequently review antiemetic treatment options and continually monitor therapeutic progress to establish the optimal therapy for each patient. PMID- 17108150 TI - Overexpression of ephrinB2 and EphB4 in tumor advancement of uterine endometrial cancers. AB - BACKGROUND: The ligand ephrinB2 and the corresponding receptor EphB4 contribute to tumor growth in various human tumors. This prompted us to study the expression and localization of ephrinB2 and EphB4 in uterine endometrial cancers to analyze the ephrinB2/EphB4 functions against clinical backgrounds. MATERIALS AND METHODS: We carried out immunohistochemistry and real-time RT-PCR to determine the histoscores and messenger RNA (mRNA) levels of ephrinB2 and EphB4, respectively, in 68 uterine endometrial cancers and 16 normal endometrium tissue samples. Patient prognoses were analyzed with a 60-month survival rate. RESULTS: The localization of ephrinB2 and EphB4 was dominantly in the cancer cells of uterine endometrial cancer of all cases given. EphrinB2 and EphB4 histoscores were highly correlated with ephrinB2 and EphB4 mRNA levels, respectively (r = 0.864 and r = 0.615, P < 0.01). Both the histoscores and mRNA levels of ephrinB2 and EphB4 significantly increased with clinical stages (I < II < III, P < 0.01), dedifferentiation (G(1) < G(2) < G(3), P < 0.01) and myometrial invasion (A < B < C, P < 0.01 for ephrinB2 and P < 0.05 for EphB4) in uterine endometrial cancers. The 60-month survival rates of the 34 patients with high ephrinB2 and EphB4 expression were poor (59% and 62% respectively), while for the other 34 patients with low ephrinB2 and EphB4 expression, they were significantly higher (85% and 82%, respectively). CONCLUSIONS: EphrinB2 and EphB4 were overexpressed during the tumor advancement as dedifferentiation and myometrial invasion. Therefore, ephrinB2/EphB4 might work on tumor advancement and may be recognized as a novel prognostic indicator for uterine endometrial cancers. PMID- 17108151 TI - Second-line chemotherapy with pegylated liposomal doxorubicin and carboplatin is highly effective in patients with advanced ovarian cancer in late relapse: a GINECO phase II trial. AB - BACKGROUND: Platinum-based chemotherapy is standard second-line treatment of patients with advanced ovarian cancer (AOC) in late relapse. Pegylated liposomal doxorubicin (PLD) has significant single-agent activity in this setting. Therefore, we evaluated the use of PLD plus carboplatin in this patient population. PATIENTS AND METHODS: PLD 30 mg/m(2) followed by carboplatin at area under the curve (AUC) 5 mg.min/ml, repeated every 28 days for a maximum of nine cycles, was administered to 104 women with AOC relapsing >or=6 months after completion of first- or second-line therapy with platinum-taxane-based regimens. RESULTS: Overall response was 63%, with a 38% complete response, median progression-free survival of 9.4 months, and median overall survival (OS) of 32 months. Grade 3 or 4 neutropenia occurred in 51% of patients, but febrile neutropenia in only 3%. Nonhematologic toxic effects were primarily grades 1 and 2, with low rates of alopecia and neurotoxicity. CONCLUSIONS: PLD plus carboplatin is highly effective, prolongs OS, and is well tolerated in women with AOC in late relapse previously treated with both platinum and taxanes. Evaluation of this regimen in phase III trials is warranted. PMID- 17108152 TI - Plastid biogenesis, between light and shadows. AB - Plastids are cellular organelles which originated when a photosynthetic prokaryote was engulfed by the eukaryotic ancestor of green and red algae and land plants. Plastids have diversified in plants from their original function as chloroplasts to fulfil a variety of other roles in metabolite biosynthesis and in storage, or purely to facilitate their own transmission, according to the cell type that harbours them. Therefore cellular development and plastid biogenesis pathways must be closely intertwined. Cell biological, biochemical, and genetic approaches have generated a large body of knowledge on a variety of plastid biogenesis processes. A brief overview of the components and functions of the plastid genetic machinery, the plastid division apparatus, and protein import to and targeting inside the organelle is presented here. However, key areas in which our knowledge is still surprisingly limited remain, and these are also discussed. Chloroplast-defective mutants suggest that a substantial number of important plastid biogenesis proteins are still unknown. Very little is known about how different plastid types differentiate, or about what mechanisms co-ordinate cell growth with plastid growth and division, in order to achieve what is, in photosynthetic cells, a largely constant cellular plastid complement. Further, it seems likely that major, separate plastid and chloroplast 'master switches' exist, as indicated by the co-ordinated gene expression of plastid or chloroplast specific proteins. Recent insights into each of these developing areas are reviewed. Ultimately, this information should allow us to gain a systems-level understanding of the plastid-related elements of the networks of plant cellular development. PMID- 17108153 TI - Effects of toluene exposure during brain growth spurt on GABA(A) receptor mediated functions in juvenile rats. AB - Toluene is a commonly abused inhalant. Its neurobiological effects are, at least in part, mediated by gamma-aminobutyric acid (GABA(A)) receptors. Since GABA(A) receptor function is critical during brain development, the long-term effects of toluene exposure during brain growth spurt were investigated. Spargue-Dawley male rats were administered with toluene (500 mg/kg, i.p.) on postnatal day (PN) 4-9. Behavioral and electrophysiological measures and the levels of messenger RNA (mRNA) expression of GABA(A) receptor subunits were examined on PN 28-32. The seizure sensitivity induced by bicuculline (a GABA(A) receptor antagonist), methyl beta-carboline-3-carboxylate (inverse agonists of the GABA(A)/benzodiazepine receptor) but not 3-mercaptopropionic acid (a glutamate decarboxylase inhibitor) was enhanced by toluene exposure. Toluene exposure had no effect on the performance in the elevated plus-maze and rotarod test but reduced the responses to diazepam in these two tests. In vitro intracellular electrophysiological recordings employing brain slices from rats treated with toluene demonstrated a significant decrease in GABA(A) receptor-mediated inhibitory postsynaptic currents in CA1 neurons but an increased response to GABA perfusion. The relative abundance of the mRNAs encoding various subunits of GABA(A) receptor (alpha1, alpha2, alpha4, alpha5, alpha6, beta2, beta3, gamma2S, gamma2L) was examined in four brain regions (hippocampus, striatum, cortex, and cerebellum) by semiquantitative reverse transcription-PCR. These results demonstrated that subunit- and brain area-selective alterations in GABA(A) receptors after toluene exposure during brain growth spurt. The alterations in GABA(A) receptors might be associated with the neurobehavioral disturbance in offspring of toluene-abusing women. PMID- 17108154 TI - The role of protein synthesis in striatal long-term depression. AB - Long-term depression (LTD) at the corticostriatal synapse is postsynaptically induced but presynaptically expressed, the depression being a result of retrograde endocannabinoid signaling that activates presynaptic cannabinoid CB1 receptors and reduces the probability of glutamate release. To study the role of protein synthesis in striatal LTD, we used a striatum-only preparation in which the presynaptic cell body is cut off, leaving intact only its axons, whose terminals synapse on medium spiny neurons. LTD (duration >150 min) was induced in this preparation, thus providing evidence that transcription in the presynaptic cell nucleus is not necessary for this form of plasticity. The maintenance of striatal LTD, however, was blocked by bath application of protein translation inhibitors but not by the same inhibitors loaded into the postsynaptic cell. These results suggest that local translation is critical for the expression of striatal LTD, distinguishing this form of mammalian synaptic plasticity from other forms that require postsynaptic protein synthesis. Possible roles of axonal or glial translation in striatal LTD are considered. PMID- 17108155 TI - Mapping iso-orientation columns by contrast agent-enhanced functional magnetic resonance imaging: reproducibility, specificity, and evaluation by optical imaging of intrinsic signal. AB - Activation resembling ocular dominance or orientation columns has been mapped with high-resolution functional magnetic resonance imaging (fMRI). However, the neuronal interpretation of these functional maps is unclear because of the poor sensitivity of fMRI, unknown point spread function (PSF), and lack of comparison with independent techniques. Here we show that cerebral blood volume (CBV) weighted fMRI with a blood plasma contrast agent (monocrystalline iron oxide nanoparticles), in combination with continuous temporally encoded stimulation, can map columnar neuronal activity in the cat primary visual cortex with high sensitivity, selectivity, and reproducibility. We examined hemodynamic response PSF by comparing these CBV-based signals with oxygen metabolism-based negative blood oxygenation level-dependent signals. A significant positive correlation exists between CBV- and metabolism-based iso-orientation maps, suggesting that the hemodynamic PSF is narrower than intercolumn distances. We also compared CBV based fMRI with optical intrinsic signal (OIS) imaging, a technique that identifies sites of increased neuronal activity, to investigate neuronal correlation. Iso-orientation maps obtained by fMRI and OIS were well matched, indicating that areas of the highest orientation-selective CBV signals correspond to sites of increased neural activity. Using CBV-based fMRI, we successfully mapped orientation-selective functional architecture in the medial bank of the visual cortex, an area inaccessible to OIS imaging. Thus, we conclude that contrast agent-based fMRI, in combination with continuous temporally encoded stimulation, is a highly sensitive technique capable of mapping neural activity at the resolution of functional columns without depth limitation. PMID- 17108156 TI - A limited access compartment between the pore domain and cytosolic domain of the BK channel. AB - Cytosolic N-terminal segments of many K+ channel subunits mediate rapid blockade of ion permeation by physical occlusion of the ion-conducting pore. For some channels with large cytosolic structures, access to the channel pore by inactivation domains may occur through lateral entry pathways or "side portals" that separate the pore domain and associated cytosolic structures covering the axis of the permeation pathway. However, the extent to which side portals control access of molecules to the channel or influence channel gating is unknown. Here we use removal of inactivation by trypsin as a tool to examine basic residue accessibility in both the N terminus of the native auxiliary beta2 subunit of Ca2+-activated, BK-type K+ channels and beta2 subunits with artificial inactivating N termini. The results show that, for BK channels, side portals define a protected space that precedes the channel permeation pathway and excludes small proteins such as trypsin but allows inactivation domains to enter. When channels are closed, inactivation domains readily pass through side portals, with a central antechamber preceding the permeation pathway occupied by an inactivation domain approximately half of the time under resting conditions. The restricted volume of the pathway through side portals is likely to influence kinetic properties of inactivation mechanisms, blockade by large pharmacological probes, and accessibility of modulatory factors to surfaces of the channel within the protected space. PMID- 17108157 TI - The development of cortical multisensory integration. AB - Although there are many perceptual theories that posit particular maturational profiles in higher-order (i.e., cortical) multisensory regions, our knowledge of multisensory development is primarily derived from studies of a midbrain structure, the superior colliculus. Therefore, the present study examined the maturation of multisensory processes in an area of cat association cortex [i.e., the anterior ectosylvian sulcus (AES)] and found that these processes are rudimentary during early postnatal life and develop only gradually thereafter. The AES comprises separate visual, auditory, and somatosensory regions, along with many multisensory neurons at the intervening borders between them. During early life, sensory responsiveness in AES appears in an orderly sequence. Somatosensory neurons are present at 4 weeks of age and are followed by auditory and multisensory (somatosensory-auditory) neurons. Visual neurons and visually responsive multisensory neurons are first seen at 12 weeks of age. The earliest multisensory neurons are strikingly immature, lacking the ability to synthesize the cross-modal information they receive. With postnatal development, multisensory integrative capacity matures. The delayed maturation of multisensory neurons and multisensory integration in AES suggests that the higher-order processes dependent on these circuits appear comparatively late in ontogeny. PMID- 17108158 TI - Massive and specific dysregulation of direct cortical input to the hippocampus in temporal lobe epilepsy. AB - Epilepsy affects 1-2% of the population, with temporal lobe epilepsy (TLE) the most common variant in adults. Clinical and experimental studies have demonstrated hippocampal involvement in the seizures underlying TLE. However, identification of specific functional deficits in hippocampal circuits associated with possible roles in seizure generation remains controversial. Significant attention has focused on anatomic and cellular alterations in the dentate gyrus. The dentate gyrus is a primary gateway regulating cortical input to the hippocampus and, thus, a possible contributor to the aberrant cortical hippocampal interactions underlying the seizures of TLE. Alternate cortical pathways innervating the hippocampus might also contribute to seizure initiation. Despite this potential importance in TLE, these pathways have received little study. Using simultaneous voltage-sensitive dye imaging and patch-clamp recordings in slices from animals with epilepsy, we assessed the relative degree of synaptic excitation activated by multiple cortical inputs to the hippocampus. Surprisingly, dentate gyrus-mediated regulation of the relay of cortical input to the hippocampus is unchanged in epileptic animals, and input via the Schaffer collaterals is actually decreased despite reduction in Schaffer-evoked inhibition. In contrast, a normally weak direct cortical input to area CA1 of hippocampus, the temporoammonic pathway, exhibits a TLE-associated transformation from a spatially restricted, highly regulated pathway to an excitatory projection with >10-fold increased effectiveness. This dysregulated temporoammonic pathway is critically positioned to mediate generation and/or propagation of seizure activity in the hippocampus. PMID- 17108159 TI - Intrinsic ON responses of the retinal OFF pathway are suppressed by the ON pathway. AB - Parallel ON and OFF pathways conduct visual signals from bipolar cells in the retina to higher centers in the brain. ON responses are thought to originate by exclusive use of metabotropic glutamate receptor 6 (mGluR6) expressed in retinal ON bipolar cells. Paradoxically, we find ON responses in retinal ganglion cells of mGluR6-null mice, but they occur at long latency. The long-latency ON responses are not blocked by metabotropic glutamate or cholinergic receptor antagonists and are not produced by activation of receptive field surrounds. We show that these longer-latency ON responses are initiated in the OFF pathways. Our results expose a previously unrecognized intrinsic property of OFF retinal pathways that generates responses to light onset. In mGluR6-null mice, long latency ON responses are observed in the visual cortex, indicating that they can be conducted reliably to higher visual areas. In wild-type (WT) mice, APB (DL-2 amino-4-phosphonobutyric acid), an mGluR6 agonist, blocks normal, short-latency ON responses but unmasks longer-latency ones. We find that these potentially confusing ON responses in the OFF pathway are actively suppressed in WT mice via two pharmacologically separable retinal circuits that are activated by the ON system in the retina. Consequently, we propose that a major function of the signaling of the ON pathway to the OFF pathway is suppression of these mistimed, and therefore inappropriate, light-evoked responses. PMID- 17108160 TI - High sensitivity to neuromodulator-activated signaling pathways at physiological [K+] of confocally imaged respiratory center neurons in on-line-calibrated newborn rat brainstem slices. AB - The pre-Botzinger complex (PBC) inspiratory center remains active in a transverse brainstem slice. Such slices are studied at high (8-10 mM) superfusate [K+], which could attenuate the sensitivity of the PBC to neuromodulators such as opiates. Findings may also be confounded because slice boundaries, drug injection sites, or location of rhythmogenic interneurons are rarely verified histologically. Thus, we first generated PBC slices with defined boundaries using novel "on-line histology" based on our finding that rostrocaudal extensions of brainstem respiratory marker nuclei are constant in newborn rats between postnatal days 0-4. At physiological superfusate [K+] (3 mM), 500- and 600-microm thick slices with the PBC in the center and the caudal boundary 0.70 and 0.76 mm caudal to the facial motonucleus generated rhythm for >2 and approximately 4 h, respectively. Rhythm was abolished by low nanomolar concentrations of the mu opiate receptor agonist DAMGO ([D-Ala2, N-Me-Phe4, Gly5-ol]enkephalin). After spontaneous arrest of bursting, rhythm was reactivated at clinically relevant or physiological concentrations by 3,5-dihydroxyphenylglycine, thyrotropin-releasing hormone, or rolipram, each affecting distinct second-messenger pathways. Two photon/confocal Ca2+ imaging revealed that these agents reactivated the same PBC neurons initially active in 3 mM [K+]. The data show that "calibrated" PBC slices at physiological [K+] generate rhythm with a high sensitivity to neuromodulators for extended time periods, whereas spontaneous "in vitro apnea" is an important tool to study the interaction of signaling pathways that modulate rhythm. Our approaches and findings provide the basis for a pharmacological and structure function analysis of the isolated respiratory center in a histologically well defined substrate at physiological [K+]. PMID- 17108161 TI - GABAergic excitation in the basolateral amygdala. AB - GABA-containing interneurons are a diverse population of cells whose primary mode of action in the mature nervous system is inhibition of postsynaptic target neurons. Using paired recordings from parvalbumin-positive interneurons in the basolateral amygdala, we show that, in a subpopulation of interneurons, single action potentials in one interneuron evoke in the postsynaptic interneuron a monosynaptic inhibitory synaptic current, followed by a disynaptic excitatory glutamatergic synaptic current. Interneuron-evoked glutamatergic events were blocked by antagonists of either AMPA/kainate or GABA(A) receptors, and could be seen concurrently in both presynaptic and postsynaptic interneurons. These results show that single action potentials in a GABAergic interneuron can drive glutamatergic principal neurons to threshold, resulting in both feedforward and feedback excitation. In interneuron pairs that both receive glutamatergic inputs after an interneuron spike, electrical coupling and bidirectional GABAergic connections occur with a higher probability relative to other interneuron pairs. We propose that this form of GABAergic excitation provides a means for the reliable and specific recruitment of homogeneous interneuron networks in the basal amygdala. PMID- 17108162 TI - Intermittent practice facilitates stable motor memories. AB - Humans adaptively control reaching movements to maintain good performance in the presence of novel forces acting on the arm. A recent study suggested that motor memories of different force conditions are not transformed from fragile to stable states, but rather are always vulnerable to interference from newly learned conditions (Caithness et al., 2004). This is contrary to the results of previous studies (Brashers-Krug et al., 1996; Shadmehr and Brashers-Krug, 1997), although all of these studies followed similar methods. Here, we show that a seemingly insignificant and inconsistently applied methodological detail may reconcile this discrepancy. Catch trials, in which the novel forces are removed, may be randomly interspersed among the more frequent force trials to assess how a subject is learning to predict the pattern of forces. In the absence of an interfering condition, subjects retained their learning until retest a day later regardless of whether they experienced catch trials. But in the presence of an interfering condition, only the subjects who had experienced forces intermittently retained their learning and thereby showed resistance to the interference. Thus, intermittent rather than constant practice conditions appear to be critical for dynamic motor memory stabilization. PMID- 17108163 TI - Cranial visceral afferent pathways through the nucleus of the solitary tract to caudal ventrolateral medulla or paraventricular hypothalamus: target-specific synaptic reliability and convergence patterns. AB - Cranial visceral afferents activate central pathways that mediate systemic homeostatic processes. Afferent information arrives in the brainstem nucleus of the solitary tract (NTS) and is relayed to other CNS sites for integration into autonomic responses and complex behaviors. Little is known about the organization or nature of processing within NTS. We injected fluorescent retrograde tracers into two nuclei to identify neurons that project to sites involved in autonomic regulation: the caudal ventrolateral medulla (CVLM) or paraventricular nucleus of the hypothalamus (PVN). We found distinct differences in synaptic connections and performance in the afferent path through NTS to these neurons. Anatomical studies using confocal and electron microscopy found prominent, primary afferent synapses directly on somata and dendrites of CVLM-projecting NTS neurons identifying them as second-order neurons. In brainstem slices, afferent activation evoked large, constant latency EPSCs in CVLM-projecting NTS neurons that were consistent with the precise timing and rare failures of monosynaptic contacts on second-order neurons. In contrast, most PVN-projecting NTS neurons lacked direct afferent input and responded to afferent stimuli with highly variable, intermittently failing synaptic responses, indicating polysynaptic pathways to higher-order neurons. The afferent-evoked EPSCs in most PVN-projecting NTS neurons were smaller and unreliable but also often included multiple, convergent polysynaptic responses not observed in CVLM-projecting neurons. A few PVN-projecting NTS neurons had monosynaptic EPSC characteristics. Together, we found that cranial visceral afferent pathways are structured distinctly within NTS depending on the projection target. Such, intra-NTS pathway architecture will substantially impact performance of autonomic or neuroendocrine reflex arcs. PMID- 17108164 TI - Expression of mutated mouse myocilin induces open-angle glaucoma in transgenic mice. AB - We developed a genetic mouse model of open-angle glaucoma by expression of mutated mouse myocilin (Myoc) in transgenic (Tg) mice. The Tyr423His point mutation, corresponding to the severe glaucoma-causing Tyr437His mutation in the human MYOC gene, was introduced into bacterial artificial chromosome DNA encoding the full-length mouse Myoc gene and long flanking regions. Both wild-type (Wt) and Tg animals expressed Myoc in tissues of the irido-corneal angle and the sclera. Expression of mutated Myoc induced the accumulation of Myoc in cell cytoplasm and prevented its secretion into the extracellular space. The levels of ATPase-1 were reduced in the irido-corneal angle of Tg mice compared with Wt animals. Tg mice demonstrated a moderate elevation of intraocular pressure, the loss of approximately 20% of the retinal ganglion cells (RGCs) in the peripheral retina, and axonal degeneration in the optic nerve. RGC depletion was associated with the shrinkage of their nuclei and DNA fragmentation in the peripheral retina. Pathological changes observed in the eyes of Tg mice are similar to those observed in glaucoma patients. PMID- 17108165 TI - Alpha-synuclein overexpression in PC12 and chromaffin cells impairs catecholamine release by interfering with a late step in exocytosis. AB - Alpha-synuclein (alpha-syn), a protein implicated in Parkinson's disease pathogenesis, is a presynaptic protein suggested to regulate transmitter release. We explored how alpha-syn overexpression in PC12 and chromaffin cells, which exhibit low endogenous alpha-syn levels relative to neurons, affects catecholamine release. Overexpression of wild-type or A30P mutant alpha-syn in PC12 cell lines inhibited evoked catecholamine release without altering calcium threshold or cooperativity of release. Electron micrographs revealed that vesicular pools were not reduced but that, on the contrary, a marked accumulation of morphologically "docked" vesicles was apparent in the alpha-syn-overexpressing lines. We used amperometric recordings from chromaffin cells derived from mice that overexpress A30P or wild-type (WT) alpha-syn, as well as chromaffin cells from control and alpha-syn null mice, to determine whether the filling of vesicles with the transmitter was altered. The quantal size and shape characteristics of amperometric events were identical for all mouse lines, suggesting that overexpression of WT or mutant alpha-syn did not affect vesicular transmitter accumulation or the kinetics of vesicle fusion. The frequency and number of exocytotic events per stimulus, however, was lower for both WT and A30P alpha-syn-overexpressing cells. The alpha-syn-overexpressing cells exhibited reduced depression of evoked release in response to repeated stimuli, consistent with a smaller population of readily releasable vesicles. We conclude that alpha syn overexpression inhibits a vesicle "priming" step, after secretory vesicle trafficking to "docking" sites but before calcium-dependent vesicle membrane fusion. PMID- 17108166 TI - Intracranial adeno-associated virus-mediated delivery of anti-pan amyloid beta, amyloid beta40, and amyloid beta42 single-chain variable fragments attenuates plaque pathology in amyloid precursor protein mice. AB - Accumulation of amyloid beta protein (Abeta) aggregates is hypothesized to trigger a pathological cascade that causes Alzheimer's disease (AD). Active or passive immunizations targeting Abeta are therefore of great interest as potential therapeutic strategies. We have evaluated the use of recombinant anti Abeta single-chain variable fragments (scFvs) as a potentially safer form of anti Abeta immunotherapy. We have generated and characterized three anti-Abeta scFvs that recognize Abeta 1-16, Abeta x-40, or Abeta x-42. To achieve widespread brain delivery, constructs expressing these anti-Abeta scFvs were packaged into adeno associated virus (AAV) vectors and injected into the ventricles of postnatal day 0 (P0) amyloid precursor protein CRND8-transgenic mice. Intracranial delivery of AAV to neonatal mice resulted in widespread neuronal delivery. In situ expression of each of the anti-Abeta scFvs after intracerebroventricular AAV serotype 1 delivery to P0 pups decreased Abeta deposition by 25-50%. These data suggest that intracranial anti-Abeta scFv expression is an effective strategy to attenuate amyloid deposition. As opposed to transgenic approaches, these studies also establish a "somatic brain transgenic" paradigm to rapidly and cost-effectively evaluate potential modifiers of AD-like pathology in AD mouse models. PMID- 17108167 TI - Stable rhodopsin/arrestin complex leads to retinal degeneration in a transgenic mouse model of autosomal dominant retinitis pigmentosa. AB - Over 100 rhodopsin mutation alleles have been associated with autosomal dominant retinitis pigmentosa (ADRP). These mutations appear to cause photoreceptor cell death through diverse molecular mechanisms. We show that K296E, a rhodopsin mutation associated with ADRP, forms a stable complex with arrestin that is toxic to mouse rod photoreceptors. This cell death pathway appears to be conserved from flies to mammals. A genetics approach to eliminate arrestin unmasked the constitutive activity of K296E and caused photoreceptor cell death through a transducin-dependent mechanism that is similar to light damage. Expressing K296E in the arrestin/transducin double knock-out background prevented transducin signaling and led to substantially improved retinal morphology but did not fully prevent cell death caused by K296E. The adverse effect of K296E in the arrestin/transducin knock-out background can be mimicked by constant exposure to low light. Furthermore, we found that arrestin binding causes K296E to mislocalize to the wrong cellular compartment. Accumulation of stable rhodopsin/arrestin complex in the inner segment may be an important mechanism for triggering the cell death pathway in the mammalian photoreceptor cell. PMID- 17108168 TI - Neural coding by two classes of principal cells in the mouse piriform cortex. AB - The piriform (or primary olfactory) cortex is a trilaminar structure that is the first cortical destination of olfactory information, receiving monosynaptic input from the olfactory bulb. Here, we show that the main input layer of the piriform cortex, layer II, is dominated by two classes of principal neurons, superficial pyramidal (SP) and semilunar (SL) cells, with strikingly different properties. Action potentials in SP cells are followed by a Ni2+-sensitive afterdepolarization that promotes burst firing, whereas SL cells fire nonbursting action potentials that are followed by a powerful afterhyperpolarization. Synaptic inputs from the olfactory bulb onto SP cells exhibit prominent paired pulse facilitation, which is attributable to residual presynaptic Ca2+ and a low probability of neurotransmitter release. In contrast, the same inputs onto SL cells do not facilitate. These distinctive synaptic and firing properties cause SP and SL cells to respond differently to in vivo-like bursts of afferent stimulation: SP cells tend to fire bursts of output action potentials at a higher frequency than the input, whereas SL cells tend to fire at a lower frequency than the input. When connected together in the canonical circuit of the piriform cortex, SP and SL cells transform the pattern of synaptic inputs they receive from the olfactory bulb, dispersing the firing rate and latency of output action potentials to an extent that depends on the strength of the input. Thus, the presence of two types of principal cells in layer II of the piriform cortex may underlie coding strategies used for the representation of odors. PMID- 17108169 TI - Growth factor treatment and genetic manipulation stimulate neurogenesis and oligodendrogenesis by endogenous neural progenitors in the injured adult spinal cord. AB - Neurons and oligodendrocytes are highly vulnerable to various insults, and their spontaneous replacement occurs to only a limited extent after damage in the adult spinal cord. The environment of injured tissue is thus thought to restrict the regenerative capacity of endogenous neural stem/progenitor cells; strategies for overcoming such restrictions remain to be developed. Here, we combined growth factor treatment and genetic manipulation to stimulate neurogenesis and oligodendrogenesis by endogenous progenitors in vivo. The recombinant retrovirus pMXIG, which was designed to coexpress green fluorescent proteins (GFPs) and a neurogenic/gliogenic transcription factor, was directly injected into the injured spinal cord parenchyma to manipulate proliferative cells in situ. We found that cells expressing Olig2, Nkx2.2, and NG2 were enriched among virus-infected, GFP positive (GFP+) cells. Moreover, a fraction of GFP+ cells formed neurospheres and differentiated into neurons, astrocytes, and oligodendrocytes in vitro, demonstrating that GFP retroviruses indeed infected endogenous neural progenitors in vivo. Neuronal differentiation of control virus-infected cells did not occur at a detectable level in the injured spinal cord. We found, however, that direct administration of fibroblast growth factor 2 and epidermal growth factor into lesioned tissue could induce a significant fraction of GFP-labeled cells to express immature neuronal markers. Moreover, retrovirus-mediated overexpression of the basic helix-loop-helix transcription factors Neurogenin2 and Mash1, together with growth factor treatment, enhanced the production and maturation of new neurons and oligodendrocytes, respectively. These results demonstrate that endogenous neural progenitors can be manipulated to replace neurons and oligodendrocytes lost to insults in the injured spinal cord. PMID- 17108170 TI - Physiologic regulation of a tetrodotoxin-sensitive sodium influx that mediates a slow afterdepolarization potential in gonadotropin-releasing hormone neurons: possible implications for the central regulation of fertility. AB - The brain controls fertility through release of gonadotropin-releasing hormone (GnRH), but the mechanisms underlying action potential patterning and GnRH release are not understood. We investigated whether GnRH neurons exhibit afterdepolarizing potentials (ADPs) and whether these are modified by reproductive state. Whole-cell current-clamp recordings of GnRH neurons in brain slices from ovariectomized mice revealed a slow ADP (sADP) after action potentials generated by brief current injection. Generating two or four spikes enhanced sADP amplitude and duration. sADP amplitude was not affected by blocking selected neurotransmitter/neuromodulator receptors, delayed-rectifier potassium channels, calcium-dependent cation channels, or hyperpolarization-activated cation channels but was halved by the calcium channel blocker cadmium and abolished by tetrodotoxin. Cadmium also reduced peak latency. Intrinsic mechanisms underlying the sADP were investigated using voltage-clamp protocols simulating action potential waveforms. A single action potential produced an inward current, which increased after double and quadruple stimulation. Cadmium did not affect current amplitude but reduced peak latency. Pretreatment with blockers of calcium-activated potassium currents (I(KCa)) reproduced this shift and blocked subsequent cadmium-induced changes, suggesting cadmium changes latency indirectly by blocking I(KCa). Tetrodotoxin abolished the inward current, suggesting that it is carried by sodium. In contrast, I(KCa) blockers increased the inward current, indicating that I(KCa) may oppose generation of the sADP. Strong sADPs were suprathreshold, generating repetitive spontaneous firing. I(ADP), sADP, and excitability were enhanced by in vivo estradiol, which triggers a preovulatory surge of GnRH release. Physiological feedback modification of this inward current and resulting sADP may modulate action potential firing and subsequent GnRH release. PMID- 17108171 TI - Suppression of the p75 neurotrophin receptor in uninjured sensory neurons reduces neuropathic pain after nerve injury. AB - The p75 neurotrophin receptor (p75NTR) has been implicated in diverse neuronal responses, including survival, cell death, myelination, and inhibition of regeneration. However, the role of p75NTR in neuropathic pain, for which there is currently no effective therapy, has not been explored. Here, we report that the pharmacological blockade of p75NTR in primary sensory neurons reversed neuropathic pain after nerve injury. Nerve injury increased the expression and axonal transport of p75NTR and phosphorylation of TrkA in the uninjured primary afferents. Functional inhibition of p75NTR suppressed injury-induced neuropathic pain and decreased the phosphorylation of TrkA and p38 mitogen-activated protein kinase, and the induction of transient receptor potential channels in dorsal root ganglion (DRG) neurons. Our results show that p75NTR induced in undamaged DRG neurons facilitates TrkA signaling and contributes to heat and cold hyperalgesia. PMID- 17108172 TI - A single mechanism can explain the speed tuning properties of MT and V1 complex neurons. AB - A recent study by Priebe et al., (2006) has shown that a small proportion (27%) of primate directionally selective, complex V1 neurons are tuned for the speed of image motion. In this study, I show that the weighted intersection mechanism (WIM) model, which was previously proposed to explain speed tuning in middle temporal neurons, can also explain the tuning found in complex V1 neurons. With the addition of a contrast gain mechanism, this model is able to replicate the effects of contrast on V1 speed tuning, a phenomenon that was recently discovered by Priebe et al., (2006). The WIM model simulations also indicate that V1 neuron spatiotemporal frequency response maps may be asymmetrical in shape and hence poorly characterized by the symmetrical two-dimensional Gaussian fitting function used by Priebe et al., (2006) to classify their cells. Therefore, the actual proportion of speed tuning among directional complex V1 cells may be higher than the 27% estimate suggested by these authors. PMID- 17108173 TI - The c-Jun N-terminal kinase activator dual leucine zipper kinase regulates axon growth and neuronal migration in the developing cerebral cortex. AB - Mammalian corticogenesis substantially depends on migration and axon projection of newborn neurons that are coordinated by a yet unidentified molecular mechanism. Dual leucine zipper kinase (DLK) induces activation of c-Jun N terminal kinase (JNK), a molecule that regulates morphogenesis in various organisms. We show here, using gene targeting in mice, that DLK is indispensable for establishing axon tracts, especially those originating from neocortical pyramidal neurons of the cerebrum. Direct and quantitative analysis of radial migration of pyramidal neurons using slice culture and a time-lapse imaging system revealed that acceleration around the subplate was affected by DLK gene disruption and by administration of a JNK inhibitor. Phosphorylation of JNK substrates, including c-Jun and doublecortin, and of JNK itself at the activation loop were partially affected in brains of DLK-deficient mouse embryos. These data suggest that DLK plays a significant role in the coordinated regulation of radial migration and axon projection by modulating JNK activity. PMID- 17108174 TI - Necdin downregulates CDC2 expression to attenuate neuronal apoptosis. AB - The cell cycle-regulatory transcription factor E2F1 induces apoptosis of postmitotic neurons in developmental and pathological situations. E2F1 transcriptionally activates many proapoptotic genes including the cyclin dependent protein kinase cell division cycle 2 (Cdc2). Necdin is a potent mitotic suppressor expressed predominantly in postmitotic neurons and interacts with E2F1 to suppress E2F1-mediated gene transcription. The necdin gene NDN is maternally imprinted and expressed only from the paternal allele. Deletion of the paternal NDN is implicated in the pathogenesis of Prader-Willi syndrome, a genomic imprinting-associated neurodevelopmental disorder. Here, we show that paternally expressed necdin represses E2F1-dependent cdc2 gene transcription and attenuates apoptosis of postmitotic neurons. Necdin was abundantly expressed in differentiated cerebellar granule neurons (CGNs). Neuronal activity deprivation elevated the expression of both E2F1 and Cdc2 in primary CGNs prepared from mice at postnatal day 6, whereas the necdin levels remained unchanged. In chromatin immunoprecipitation analysis, endogenous necdin was associated with the cdc2 promoter containing an E2F-binding site in activity-deprived CGNs. After activity deprivation, CGNs underwent apoptosis, which was augmented in those prepared from mice defective in the paternal Ndn allele (Ndn(+m/-p)). The levels of cdc2 mRNA, protein, and kinase activity were significantly higher in Ndn(+m/-p) CGNs than in wild-type CGNs under activity-deprived conditions. Furthermore, the populations of Cdc2-immunoreactive and apoptotic cells were increased in the cerebellum in vivo of Ndn(+m/-p) mice. These results suggest that endogenous necdin attenuates neuronal apoptosis by suppressing the E2F1-Cdc2 system. PMID- 17108175 TI - The biochemical and neuroendocrine bases of the hyperalgesic nocebo effect. AB - Despite the increasing research on placebos in recent times, little is known about the nocebo effect, a phenomenon that is opposite to the placebo effect and whereby expectations of symptom worsening play a crucial role. By studying experimental ischemic arm pain in healthy volunteers and by using a neuropharmacological approach, we found that verbally induced nocebo hyperalgesia was associated to hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis, as assessed by means of adrenocorticotropic hormone and cortisol plasma concentrations. Both nocebo hyperalgesia and HPA hyperactivity were antagonized by the benzodiazepine diazepam, suggesting that anxiety played a major role in these effects. The administration of the mixed cholecystokinin (CCK) type-A/B receptor antagonist proglumide blocked nocebo hyperalgesia completely but had no effect on HPA hyperactivity, which suggests a specific involvement of CCK in the hyperalgesic but not in the anxiety component of the nocebo effect. Importantly, both diazepam and proglumide did not show analgesic properties on basal pain, because they acted only on the nocebo-induced pain increase. These data indicate a close relationship between anxiety and nocebo hyperalgesia, in which the CCKergic systems play a key role in anxiety-induced hyperalgesia. These results, together with previous findings showing that placebo analgesia is mediated by endogenous opioids, suggest that the analgesic placebo/hyperalgesic nocebo phenomenon may involve the opposite activation of endogenous opioidergic and CCKergic systems. PMID- 17108176 TI - Synaptic integration of olfactory information in mouse anterior olfactory nucleus. AB - Individual odorants activate only a small fraction of mitral cells in the mouse main olfactory bulb (MOB). Odor mixtures are represented by a combination of activated mitral cells, forming reproducible activation maps in the olfactory bulb. However, how the activation of a cohort of narrowly tuned mitral cells by odor mixtures is read out synaptically by neurons in higher-level olfactory structures, such as the anterior olfactory nucleus (AON), is mostly unknown. In the current study, we used intracellular and extracellular recordings to examine and compare responses of AON neurons and MOB mitral cells to a panel of structurally diverse odorants presented either as mixtures or as individual components. We found that a majority of individual AON neurons could be synaptically activated by several mixtures of structurally dissimilar components and by several dissimilar components in an effective mixture. The suprathreshold response of an AON neuron to an effective mixture often exceeded the sum of its suprathreshold responses to all of the components in that mixture, indicating a nonlinear combinatorial interaction. In contrast to the broad responsiveness of AON neurons, the majority of mitral cells were activated by only one or two components in a single mixture. The broader responsiveness of AON neurons relative to mitral cells suggests that individual AON neurons synaptically integrate several functionally distinct mitral cell inputs. PMID- 17108177 TI - Primary afferent NMDA receptors increase dorsal horn excitation and mediate opiate tolerance in neonatal rats. AB - Repeated exposure to opiates produces analgesic tolerance, which limits their clinical usefulness. Whole-cell voltage-clamped lamina I cells in spinal slices from opiate-tolerant neonatal rats show an increase in miniature, spontaneous, and primary afferent-evoked EPSCs when compared with lamina I cells from opiate naive rat spinal slices. This increased excitation can be blocked by the NMDA receptor (NMDAR) antagonist APV, apparently acting at NMDARs on primary afferents. Consistent with these results, electron microscopy demonstrates an increased incidence of NMDARs in substance P-containing spinal dorsal horn primary afferent terminals in opiate-tolerant rats. Moreover, superfusion of spinal slices from opiate-tolerant rats with NMDA produces a reversible increase in miniature EPSC (mEPSC) frequency in contrast to a decrease in mEPSC frequency produced by NMDA in opiate-naive slices. Finally, NMDAR antagonists inhibit the expression of opiate tolerance both in inhibiting EPSCs in spinal slices and in inhibiting behavioral nociceptive responses to heat. NMDAR antagonists have been reported in many studies to inhibit morphine analgesic tolerance. Our studies suggest that an increase in primary afferent NMDAR expression and activity mediates a hypersensitivity to noxious stimuli and causes the inhibition of opiate efficacy, which defines tolerance. PMID- 17108178 TI - Boundary completion is automatic and dissociable from shape discrimination. AB - Normal visual perception readily overcomes suboptimal or degraded viewing conditions through perceptual filling-in processes, enhancing object recognition and discrimination abilities. This study used visual evoked potential (VEP) recordings in conjunction with electrical neuroimaging analyses to determine the spatiotemporal brain dynamics of boundary completion and shape discrimination processes in healthy humans performing the so-called "thin/fat" discrimination task (Ringach and Shapley, 1996) with stimuli producing illusory contours. First, results suggest that boundary completion processes occur independent of subjects' accuracy on the discrimination task. Modulation of the VEP to the presence versus absence of illusory contours [the IC effect (Murray et al., 2002)] was indistinguishable in terms of response magnitude and scalp topography over the 124-186 ms poststimulus period, regardless of whether task performance was correct. This suggests that failure on this discrimination task is not primarily a consequence of failed boundary completion. Second, the electrophysiological correlates of thin/fat shape discrimination processes are temporally dissociable from those of boundary completion, occurring during a substantially later phase of processing (approximately 330-406 ms). The earlier IC effect was unaffected by whether the perceived contour produced a thin or fat shape. In contrast, later time periods of the VEP modulated according to perceived shape only in the case of stimuli producing illusory contours, but not for control stimuli for which performance was at near-chance levels. Collectively, these data provide further support for a multistage model of object processing under degraded viewing conditions. PMID- 17108179 TI - Vesicular glutamate transporter VGLUT2 expression levels control quantal size and neuropathic pain. AB - Uptake of L-glutamate into synaptic vesicles is mediated by vesicular glutamate transporters (VGLUTs). Three transporters (VGLUT1-VGLUT3) are expressed in the mammalian CNS, with partial overlapping expression patterns, and VGLUT2 is the most abundantly expressed paralog in the thalamus, midbrain, and brainstem. Previous studies have shown that VGLUT1 is necessary for glutamatergic transmission in the hippocampus, but the role of VGLUT2 in excitatory transmission is unexplored in glutamatergic neurons and in vivo. We examined the electrophysiological and behavioral consequences of loss of either one or both alleles of VGLUT2. We show that targeted deletion of VGLUT2 in mice causes perinatal lethality and a 95% reduction in evoked glutamatergic responses in thalamic neurons, although hippocampal synapses function normally. Behavioral analysis of heterozygous VGLUT2 mice showed unchanged motor function, learning and memory, acute nociception, and inflammatory pain, but acquisition of neuropathic pain, maintenance of conditioned taste aversion, and defensive marble burying were all impaired. Reduction or loss of VGLUT2 in heterozygous and homozygous VGLUT2 knock-outs led to a graded reduction in the amplitude of the postsynaptic response to single-vesicle fusion in thalamic neurons, indicating that the vesicular VGLUT content is critically important for quantal size and demonstrating that VGLUT2-mediated reduction of excitatory drive affects specific forms of sensory processing. PMID- 17108180 TI - Precise spatial relationships between mossy fibers and climbing fibers in rat cerebellar cortical zones. AB - Classically, mossy fiber and climbing fiber terminals are regarded as having very different spatial distributions in the cerebellar cortex. However, previous anatomical studies have not studied these two major cerebellar inputs with sufficient resolution to confirm this assumption. Here, we examine the detailed pattern of collateralization of both types of cerebellar afferent using small injections of the bidirectional tracer cholera toxin b subunit into the posterior cerebellum. The cortical and zonal location of these injections was characterized by mapping climbing fiber field potentials, the distribution of retrogradely labeled olivary neurons, and the intrinsic zebrin pattern of Purkinje cells. Labeled climbing fiber collaterals were distributed as longitudinal strips and were always accompanied by clusters of labeled mossy fiber rosettes in the subjacent granular layer. Two- and three-dimensional reconstructions and quantitative analysis showed that mossy fibers also collateralized to other stripe-like regions usually below Purkinje cells with the same zebrin-positive or zebrin-negative characteristics as that of the injection site and associated climbing fiber collaterals. The distribution of retrogradely labeled neurons in two major sources of mossy fibers, the lateral reticular and basilar pontine nuclei, revealed interlobular and some interzonal differences. These data indicate that nonadjacent cerebellar zones, sharing the same climbing fiber input and zebrin identity, also share a common mossy fiber input. Other cerebellar cortical regions that receive collaterals from the same mossy fibers usually also have the same zebrin signature. Together with the distribution of neurons in precerebellar centers, the findings suggest a revision of the modular hypothesis for information processing in the cerebellar cortex. PMID- 17108181 TI - Structure of the catalytic pore of gamma-secretase probed by the accessibility of substituted cysteines. AB - Several single-span membrane proteins are cleaved within their transmembrane domains (TMDs) by intramembrane-cleaving proteases, although the structure of the active site executing intramembrane cleavage remains unknown. Here we use the substituted cysteine accessibility method to examine the structure of presenilin 1, a catalytic subunit of gamma-secretase, involved in amyloid beta protein generation in Alzheimer's disease and Notch signaling. We show that TMD6 and TMD7 of presenilin-1 contribute to the formation of a hydrophilic pore within the membrane. Residues at the luminal portion of TMD6 are predicted to form a subsite for substrate or inhibitor binding on the alpha-helix facing a hydrophilic milieu, whereas those around the GxGD catalytic motif within TMD7 are highly water accessible, suggesting formation of a hydrophilic structure within the pore. Collectively, our data suggest that the active site of gamma-secretase resides in a catalytic pore filled with water within the lipid bilayer and is tapered around the catalytic aspartates. PMID- 17108182 TI - Leukemia inhibitory factor promotes neural stem cell self-renewal in the adult brain. AB - Although neural stem cells (NSCs) persist in various areas of the adult brain, their contribution to brain repair after injury is very limited. Treatment with exogenous growth factors can mitigate this limitation, suggesting that the brain environment is normally deficient in permissive cues and that it may be possible to stimulate the latent regenerative potential of endogenous progenitors with appropriate signals. We analyzed the effects of overexpressing the cytokine leukemia inhibitory factor (LIF) on adult neurogenesis in the normal brain. We found that LIF reduces neurogenesis in the olfactory bulb and subventricular zone by acting directly on NSCs. LIF appears to promote NSC self-renewal, preventing the emergence of more differentiated cell types. This ultimately leads to an expansion of the NSC pool. Our results have implications for the development of therapeutic strategies for brain repair and suggest that LIF may be useful, in combination with other factors, in promoting regeneration in the adult brain. PMID- 17108183 TI - An analysis of 5'-inosine and 5'-guanosine monophosphate taste in rats. AB - Inosine monophosphate (IMP) and guanosine monophosphate (GMP) elicit an umami taste in humans and synergistically increase the intensity of the umami taste of monosodium glutamate (MSG). Conditioned taste aversion (CTA) studies in rodents indicate that these nucleotides and MSG elicit quite similar tastes, but recent physiological evidence suggests that these nucleotides and MSG may not activate the same population of taste receptors and therefore may not elicit identical taste qualities. This study reports the findings of several behavioral experiments with rats that compared the taste properties of IMP and GMP with each other and with those of MSG. Well-trained rats were able to detect both nucleotides at nanomolar concentrations, but they did not respond to either nucleotide in two-bottle preference tests or brief-access CTA tests at concentrations less than 0.5 mM. Discrimination experiments found that the tastes of these nucleotides could not be discriminated from each other, but both could be discriminated from MSG, even when the taste of Na(+) was controlled. Overall, these experiments indicate the taste properties of the two 5'-ribonucleotides are quite similar to each other, and even though they may elicit an umami sensation, these sensations are not identical to the taste of MSG. PMID- 17108184 TI - Dynamics of reductive genome evolution in mitochondria and obligate intracellular microbes. AB - Reductive evolution in mitochondria and obligate intracellular microbes has led to a significant reduction in their genome size and guanine plus cytosine content (GC). We show that genome shrinkage during reductive evolution in prokaryotes follows an exponential decay pattern and provide a method to predict the extent of this decay on an evolutionary timescale. We validated predictions by comparison with estimated extents of genome reduction known to have occurred in mitochondria and Buchnera aphidicola, through comparative genomics and by drawing on available fossil evidences. The model shows how the mitochondrial ancestor would have quickly shed most of its genome, shortly after its incorporation into the protoeukaryotic cell and prior to codivergence subsequent to the split of eukaryotic lineages. It also predicts that the primary rickettsial parasitic event would have occurred between 180 and 425 million years ago (MYA), an event of relatively recent evolutionary origin considering the fact that Rickettsia and mitochondria evolved from a common alphaproteobacterial ancestor. This suggests that the symbiotic events of Rickettsia and mitochondria originated at different time points. Moreover, our model results predict that the ancestor of Wigglesworthia glossinidia brevipalpis, dated around the time of origin of its symbiotic association with the tsetse fly (50-100 MYA), was likely to have been an endosymbiont itself, thus supporting an earlier proposition that Wigglesworthia, which is currently a maternally inherited primary endosymbiont, evolved from a secondary endosymbiont. PMID- 17108185 TI - Linkage between 'disruption of inactivation' and 'reduction of K+ selectivity' among hERG mutants in the S5-P linker region. PMID- 17108186 TI - Circulating nucleic acids in plasma and serum: recent developments. AB - Nucleic acids (DNA and RNA) have been detected in plasma, serum, urine, and other body fluids from healthy subjects as well as in patients. The ability to detect and quantitate specific DNA and RNA sequences has opened up the possibility of diagnosis and monitoring of diseases. With the recent developments in the field of circulating nucleic acids the application in the diagnostic field has increased. The recent discovery of epigenetic changes in placental/fetal DNA and the detection of fetal/placental-specific RNAs have made it possible to use this technology in all pregnancies irrespective of the gender of the fetus. With the application of mass spectrometry and other techniques to this field, it is now possible to detect very small amounts of specific DNA in the presence of excess of other nonspecific nucleic acids (e.g., detection of mutations in fetal DNA in the presence of excess of maternal DNA). Circulating nucleic acids have now been shown to be useful in other conditions, such as diabetes mellitus, trauma, stroke, and myocardial infarction. In oncology, detection and monitoring of tumors is now possible by the detection of tumor-derived nucleic acids. In spite of these advances questions regarding the origin and biologic significance of circulating nucleic acids remain to be answered. Furthermore pre-analytical and analytical aspects of this field remain to be standardized. PMID- 17108187 TI - Prehistory of the notion of circulating nucleic acids in plasma/serum (CNAPS): birth of a hypothesis. AB - In the late 50s and early 60s of the last century, a theoretical fight was taking place between Western and Russian scientists about the theory explaining the mechanism of evolution. According to neo-Darwinism, evolution was the result of hazard and necessity, that is, mutations arriving by chance favoring the survival of the fittest. For the Russian geneticists, acquired characteristics were the basis of evolution, that is, the environment modified the characteristics of the gene. One of the main experiments on which the Russian geneticists based their theory was the transmission of hereditary characteristics by a special technique of grafting between two varieties of plants-a mentor plant and a pupil plant. The pupil variety being entirely dependent on the development of the mentor plant its hereditary characteristics were modified accordingly. In the Western world these experiments were regarded with doubt. We were among the few who tried to repeat this kind of experiment. After three generations of grafting between two varieties of eggplant, we succeeded in obtaining hereditary modifications of the pupil plants, which acquired some of the characteristics of the mentor variety. The linkage between some hereditary characteristics of the mentor plant were broken, the segregation of the offspring was abnormal, dominant characteristics appearing in the offspring of a recessive plant. Rather than adopting the views of the Russian scientists about acquired characteristics, we suggested that DNA was circulating between the mentor and pupil plants and assumed that some nucleic acid molecules bearing genetic information could enter the somatic and reproductive cells of the pupil plant at a propitious moment and remain active. PMID- 17108188 TI - Circulating DNA: intracellular and intraorgan messenger? AB - The circulation of both foreign and endogenous DNA within plants and its ability to be expressed in the host plants and FI generation is described. These data, together with those from animal systems are used to support the concept that a DNA fraction can act as a messenger between cells and tissues. PMID- 17108189 TI - Immunological aspects of circulating DNA. AB - Nude mice were injected with DNA released by T lymphocytes previously exposed to inactivated herpes symplex type 1 or polio viruses. The serum of these mice was tested for its neutralizing activity. Injected nude mice synthesized antiherpetic or antipolio antibodies, depending on the antigen used to sensitize the T lymphocytes. Mice injected with DNA released by human T cells produced antibodies carrying human allotypes as they could be neutralized by antiallotype sera. However, mice that were injected with DNA released by antigen-stimulated murine T lymphocytes produced antiviral antibodies, which were not neutralized by anti human allotype sera. PMID- 17108190 TI - Biology of circulating mRNA: still more questions than answers? AB - A few years after the first description of free-circulating DNA in plasma and serum, the detection of tumor-associated overexpressed mRNA in plasma was also reported. This observation has been confirmed and it seems to be clear that the presence of free-circulating RNA is a ubiquitous phenomenon. In this short review I will discuss some basic aspects of the release mechanisms for the RNA, its biological meaning, and clinical value. PMID- 17108191 TI - Investigation of the origin of extracellular RNA in human cell culture. AB - We have used the human ECV 304 cell line to study the origin and fate of extracellular RNA (exRNA) in cell culture. Quantification of different extracellular RNA species using reverse transcription followed by quantitative PCR revealed a prevalent fraction of ribosomal RNAs. Comparison of intracellular and extracellular ribosomal RNA copy numbers allowed the calculation of the number of destroyed cells that would result in the corresponding number of extracellular rRNAs. Interestingly, this number was comparable to the amount of destroyed cells as determined by the measurement of extracellular lactate dehydrogenase activity. PMID- 17108192 TI - Hypoxia-induced membrane-bound apoptotic DNA particles: potential mechanism of fetal DNA in maternal plasma. AB - Fetal DNA is found in the plasma of pregnant women that appears to be stable for PCR amplification. Although the underlying mechanism giving rise to this DNA in plasma remains unclear, the source of these fragments may be from apoptotic bodies (Apo-Bodies) created from dying cells. Trophoblast apoptosis is essential for normal placental development, given the enormous amount of proliferation, differentiation, and migration during pregnancy. Through flow cytometric analysis coupled with real-time PCR, our lab has shown that aggregates of acridine orange (AO)-stained material (apoptotic particles) are resistant to DNase treatment, disrupted by sodium dodecyl sulfate (SDS), and contain fetal DNA. Because the placenta continuously re-models in an hypoxic environment, our hypothesis is that fetal DNA in maternal plasma comes from hypoxia-induced dying trophoblasts and that this DNA circulates predominantly in the form of Apo-Bodies. We have developed a model culture system for analysis of Apo-Bodies derived from JEG-3 cells, an extravillous trophoblastic cell line, undergoing various methods of cell death: hypoxia-induced, etoposide-induced, and heat stress (necrosis like) induced cell death. Under conditions of similar propidium iodide (PI) uptake, suggesting comparable levels of death, both hypoxia- and etoposide-induced Apo Bodies increase in concentration over time, whereas heat-induced levels of particles remain fairly constant, indicating that production of DNA-associated Apo-Bodies is a continuous process. Hypoxia, which is likely to be responsible for trophoblast cell death in vivo, produced membrane-bound Apo-Bodies containing DNA. Our results are consistent with the characteristics of membrane-bound particles containing fetal DNA found in maternal plasma. PMID- 17108193 TI - Fetal nucleic acids in maternal body fluids: an update. AB - Our laboratory continues to be actively involved in the development of new biomarkers for prenatal diagnosis using maternal blood and amniotic fluid. We have also developed a mouse model that demonstrates that cell-free fetal (cff) DNA is detectable in the pregnant maternal mouse. In human maternal plasma and serum we have analyzed factors that are important in the clinical interpretation of cff DNA levels. Maternal race, parity, and type of conception (natural or assisted) do not affect cff DNA levels, but maternal weight does. We have also analyzed the relationship between placental volume, using a three-dimensionsal ultrasound examination, and cff DNA levels. Surprisingly, there is no association between these values. Finally, we are using specific disease models (such as congenital diaphragmatic hernia and twin-to-twin transfusion) to understand the effects of gestational age and specific pathology on fetal gene expression by analyzing cell-free mRNA levels in maternal plasma. In the amniotic fluid we have focused on improvements in recovery of cff DNA and mRNA. By optimizing recovery we have made some interesting observations about differences in fetal DNA between blood and amniotic fluid. In addition, we have successfully hybridized cff DNA in amniotic fluid to DNA microarrays, permitting assessment of fetal molecular karyotype. We also have preliminary data on fetal gene expression in amniotic fluid. Finally, we remain actively involved in promoting noninvasive prenatal testing in the United States, such as encouraging the use of fetal DNA for fetal rhesus D assessment. On the other hand, we are cautious and concerned about the accuracy of "at-home" kits for fetal gender detection. PMID- 17108194 TI - Fetal DNA in maternal plasma: progress through epigenetics. AB - The discovery of cell-free fetal DNA in maternal plasma has opened up new possibilities for noninvasive prenatal diagnosis. Most of the work in this field has focused on the detection of fetal genetic markers that are distinguishable from the background maternal DNA. The feasibility of detecting fetal epigenetic markers in maternal plasma using an imprinted locus was demonstrated in 2002. This work has recently led to the development of the first universal fetal epigenetic marker, hypomethylated maspin, which can be used for fetal DNA detection in maternal plasma, irrespective of fetal gender and genetic polymorphisms. It is expected that many more fetal epigenetic markers that can be used in this manner will be developed over the next few years. These markers may catalyze the eventual clinical use of circulating fetal DNA for noninvasive prenatal diagnosis, such as for the detection of fetal chromosomal aneuploidies. PMID- 17108195 TI - Cell-free DNA in maternal plasma: is it all a question of size? AB - Fetal cell-free DNA (cf-DNA) represents only a small fraction of the total cf-DNA in maternal plasma. This feature has rendered it difficult to reliably distinguish fetal alleles which are not very disparate from maternal ones, such as those involving point mutations, by conventional polymerase chain reaction (PCR)-based approaches. It has recently been shown that cell-free fetal DNA molecules have a smaller size than comparable cf-DNA molecules of maternal origin, and that this feature can be exploited for the selective enrichment of fetal DNA sequences, thereby permitting the detection of otherwise masked fetal genetic traits. By the use of this approach, we have shown that it is possible to detect fetal genetic loci for microsatellite markers, as well as point mutations involved in disorders such as achondroplasia and beta-thalassemia. PMID- 17108196 TI - Fetal blood group genotyping: present and future. AB - Prediction of fetal blood group from DNA is usually performed when the mother has antibodies to RhD, to assess whether the fetus is at risk from hemolytic disease of the fetus and newborn (HDFN). Over the last five years RhD testing on fetal DNA in maternal plasma has been introduced. At the International Blood Group Reference Laboratory (IBGRL) we employ real-time quantitative polymerase chain reaction (RQ-PCR) to detect RHD exons 4, 5, and 10, which also reveals RHDpsi. SRY and, in RhD-negative (RhD-) females, eight biallelic polymorphisms are incorporated in an attempt to provide an internal positive control. Since 2000 we have tested 533 pregnancies for RhD. In 327 pregnancies where the RhD of the infant is known, we had one false-positive and one false-negative result. In 2004 we introduced fetal typing from DNA in maternal plasma for K, Rhc, and RhE, which represent single nucleotide polymorphisms (SNPs) on the KEL and RHCE genes. We have begun trials on an automated method for fetal RhD typing from DNA in maternal plasma. This is designed to test fetal RhD in all pregnant RhD- women, to identify the 40% with an RhD- fetus so that antenatal RhD immunoglobulin (Ig) prophylaxis can be avoided. Similar trials have already been reported by Sanquin Research Laboratories in Amsterdam. PMID- 17108197 TI - Placental RNA in maternal plasma: toward noninvasive fetal gene expression profiling. AB - The recent demonstration of the presence of placenta-derived fetal RNA in maternal plasma has opened up new opportunities for noninvasive prenatal investigation. Circulating fetal RNA analysis could in principle be applied to all pregnancies without the limitations by fetal gender or polymorphisms between the mother and fetus. The use of fetus- or disease-specific circulating RNA markers would greatly increase the number of markers that can be used for prenatal monitoring. With the recent advances in microarray technology, new placenta-derived plasma RNA markers could be rapidly identified. These newly identified placental transcripts were robustly detectable in maternal plasma and were pregnancy-specific. Remarkably, the relative concentrations of the placental mRNA in maternal plasma directly reflect the gene expression patterns in the placenta, an observation which suggests that placental gene expression level plays a predominant role in determining the placental mRNA concentrations in maternal plasma. Thus, fetal mRNA measurement in maternal plasma may be a useful tool for noninvasive prenatal placental gene expressing profiling. PMID- 17108198 TI - Development and application of a real-time quantitative PCR for prenatal detection of fetal alpha(0)-thalassemia from maternal plasma. AB - In order to provide a noninvasive prenatal diagnosis of alpha(0)-Thalassemia (Southeast Asian [SEA] deletion), we have developed a real-time quantitative semi nested polymerase chain reaction (PCR) method for identifying the fetal alpha(0) Thalassemia in maternal plasma. Analysis was performed using DNA extracted from 200 muL plasma from 13 pregnant women during 8-20 weeks of gestation who carried fetuses with normal (2), alpha(0)-Thalassemia carrier (8), Hb H disease (1), and homozygous alpha(0)-Thalassemia (Hb Bart's hydrops fetalis (2). The alpha(0) Thalassemia was detected using a two-step PCR. Plasma DNA was amplified conventionally using alpha(0)-Thalassemia-specific primers and a portion of the first PCR product was subjected to a semi-nested real-time q-PCR using the SYBR green I chemistry for fluorescence detection. Calibration curve for alpha(0) Thalassemia quantification was prepared by assaying serial dilution of genomic DNA of an alpha(0)-Thalassemia carrier. Differences in the C(T) (threshold cycle) values and calculated concentrations of amplified DNA among normal fetus, alpha(0)-Thalassemia carrier, Hb H disease, and homozygous alpha(0)-Thalassemia were clearly observed, which could help in prenatal prediction of the fetal genotype. This noninvasive prenatal detection of alpha(0)-Thalassemia in maternal plasma should enhance prenatal diagnostic options for this common genetic disorder in routine DNA diagnostic setting. PMID- 17108199 TI - Detection of a paternally inherited fetal mutation in maternal plasma by the use of automated sequencing. AB - The discovery of circulating fetal DNA in maternal blood has been an encouraging step forward in the prenatal diagnostic field. It has opened up the possibility of development of a noninvasive method for the genetic analysis of the fetus. Many techniques have been applied to the study of this fetal DNA, but automated sequencing has been seldom used. The intention of this study was to use the automated sequencing technique for the detection of a paternally inherited fetal mutation in maternal plasma. Maternal plasma samples from a pregnant woman, whose husband had a mutation (Q134X) in the RP2 gene, which is located in the X chromosome, were collected at two different gestational ages (10th and 19th week of gestation) in order to determine whether the paternally inherited fetal mutation could be detected by automated sequencing. Restriction analysis was also performed to confirm the results. The fetal mutation was clearly detected in the maternal plasma by the use of automated sequencing. The automated sequencing enables the possibility of analyzing fetal sequences, at a nucleotide level, in order to detect mutations or polymorphisms which are distinguishable from maternal sequences. PMID- 17108200 TI - Occurrence of neutrophil extracellular DNA traps (NETs) in pre-eclampsia: a link with elevated levels of cell-free DNA? AB - Manifest pre-eclampsia is associated with activation of peripheral neutrophils as well as elevations in maternal cell-free DNA. For this reason, we were very intrigued by recent reports indicating that activated circulatory neutrophils secrete nuclear DNA to generate extracellular DNA lattices, termed NETs (neutrophil extracellular traps). Our preliminary data indicate that placental syncytiotrophoblast microparticles, which are released in elevated amounts in pre eclampsia, can induce NETs in isolated neutrophils. Furthermore, we found evidence for the increased presence of NETs directly in the intervillous space of pre-eclamptic placentae. Therefore, these newly discovered entities may be implicated in the underlying etiology of this disorder. PMID- 17108201 TI - Use of bi-allelic insertion/deletion polymorphisms as a positive control for fetal genotyping in maternal blood: first clinical experience. AB - Amplification of fetal DNA in maternal plasma is a new way for non-invasive fetal genotyping in pregnancies at risk for disorders where the presence of a paternal DNA sequence contributes to the risk status of the fetus. We describe the use of a panel of 10 bi-allelic highly polymorphic markers to ascertain the presence and amplification of fetal DNA in case the fetus is negative for the targeted paternal "disease" sequence. PMID- 17108202 TI - PLAC1 mRNA in maternal blood correlates with Doppler waveform in uterine arteries in normal pregnancies at the second and third trimester. AB - Doppler analysis of the uterine arteries is currently used for pre-eclampsia (PE) screening. PLAC1 is a trophoblast-specific gene, and it is known that in normal pregnancies, trophoblastic cells are released into the maternal circulation, where specific trophoblastic mRNA can be detected. In PE, as in women who eventually develop PE, an abnormal passage of fetal and placental cells is also present. In this study, we aimed to verify whether, in normal pregnancies, Doppler waveform of the uterine arteries correlates with PLAC1 mRNA concentrations. Thirteen cases of normal pregnancies at 37 weeks' gestation (23 41) were enrolled in the study. PLAC1 mRNA was extracted from 2 mL of blood by ABI PRISM 6100 nucleic acid Prep Station (Applied Biosystems, Foster City, CA) and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was performed by a PE 5700 Sequence detection system. Bulk RNA from normal placental tissue was used as the reference curve, and the amount of PLAC1 mRNA in the study samples was then expressed as the "relative amount" of weight of placental tissue (ng/mL). The uterine arterial mean resistance index (RI) and presence/absence of a dicrote waveform were calculated by using a 5 MHz transabdominal probe (Tecnos, ESAOTE) at the uterine cervico-corporal junction. Doppler measurement was performed on the same day as blood collection. The median of the means of uterine arterial RI was 0.52 (0.39-0.68). RI of uterine arteries and PLAC1 mRNA were significantly correlated in a log-linear regression (R(2) = 0.483, P = 0.024). Our data support that in normal pregnancy, the passage of trophoblast material into the maternal circulation is correlated with the quantitative measurement of uterine hemodynamics. PMID- 17108203 TI - Different approaches for noninvasive prenatal diagnosis of genetic diseases based on PNA-mediated enriched PCR. AB - The aim of this work was to develop advanced and accessible protocols for noninvasive prenatal diagnosis of genetic diseases. We are evaluating different technologies for mutation detection, based on fluorescent probe hybridization of the amplified product and pyrosequencing, a technique that relies on the incorporation of nucleotides in a primer-directed polymerase extension reaction. In a previous investigation, we have already proven that these approaches are sufficiently sensitive to detect a few copies of a minority-mutated allele in the presence of an excess of wild-type DNA, In this work, in order to further enhance the sensitivity, we have employed a mutant enrichment amplification strategy based on the use of peptide nucleic acids (PNAs). These DNA analogues bind wild type DNA, thus interfering with its amplification while still allowing the mutant DNA to become detectable. We have synthesized different PNAs, which are highly effective in clamping wild-type DNA in the beta-globin gene region, where four beta-thalassemia mutations are located (IVSI.110, CD39, IVSI.1, IVSI.6) plus HbS. The fluorescence microchip readout allows us to monitor the extent of wild-type allele inhibition, thus facilitating the assessment of the optimal PNA concentration. PMID- 17108204 TI - Detection of SNPs in the plasma of pregnant women and in the urine of kidney transplant recipients by mass spectrometry. AB - Recently, it has been discovered that cell-free fetal DNA is smaller than corresponding maternal DNA. Therefore, circulating fetal DNA can be enriched by size-fractionation. Such a selection improves the non-invasive prenatal diagnosis of paternally inherited single gene mutations. Recent studies showed that MALDI TOF mass spectrometry (MS) can be used to reliably detect fetal-specific single nucleotide polymorphisms (SNPs) in maternal plasma. In this study, we looked at whether the size-fractionation approach could improve the detection of paternally inherited SNPs by MS assay. Our results indicated that the size-fractionation approach improved the analysis of paternally inherited SNP alleles. Our previous studies showed that donor-derived STR sequences could be detected in the urine of kidney transplant recipients. Here, we also examined whether donor-specific SNPs could be detected in recipient's urine by MS. PMID- 17108205 TI - Noninvasive prenatal diagnostic assay for the detection of beta-thalassemia. AB - The development of a noninvasive method for detection of beta-thalassemia in the population of Cyprus is based on the detection of paternally inherited single nucleotide polymorphisms (SNPs) as well as beta-thalassemia (beta-thal) mutations. We selected 11 informative SNPs for the Cypriot population linked to the beta-globin locus. Two different approaches were used: allele-specific polymerase chain reaction (AS-PCR) and the arrayed primer extension (APEX) method. The AS-PCR approach is being standardized, and the method was applied in two families. The paternally inherited allele was noninvasively detected with the AS-PCR approach on maternal plasma. Some preliminary tests were performed with the APEX method on genomic DNA of parents carrying the beta-thal mutation. PMID- 17108206 TI - Circulating DNA and lung cancer. AB - Lung cancer is the leading cause of cancer death worldwide. The majority of patients is diagnosed too late for curative treatment. There is an urgent need for a noninvasive test to identify early lung cancer. Although levels of circulating cell-free DNA in plasma or serum are higher in patients with lung cancer than in healthy controls, it is not yet clear whether this will be of diagnostic or prognostic significance. The finding that circulating DNA in lung cancer patients exhibits genetic and epigenetic changes typical of the tumor (including chromosome loss, oncogene activation, and tumor-suppressor gene inactivation by methylation) has led to intense efforts to determine whether these are sensitive and specific enough to be used clinically. Here we review the evidence on circulating DNA in lung cancer and consider possible future applications in patient management. PMID- 17108207 TI - Circulating nucleic acids in plasma/serum and tumor progression: are apoptotic bodies involved? An experimental study in a rat cancer model. AB - The "genometastasis hypothesis" proposes that cell-free tumor nucleic acids might be able to transform host stem cells, and that this might be a pathway for the development of metastases. This theory is supported by previous experimental findings and is consistent with observations of other authors. It has been suggested that tumor DNA might be horizontally transferred by the uptake of apoptotic bodies and initiate the genetic changes that are necessary for tumor formation. In addition, apoptotic bodies have been proposed as possible vehicles that protect the nucleic acids circulating in the plasma from enzymatic degradation. In the present study, we analyzed the presence of apoptotic bodies in serum and its relationship with tumor progression in a heterotopic model of colon cancer in the rat. We injected DHD/K12-PROb cancer cells subcutaneously into BD-IX rats and divided the animals into three groups according to the time between the injection of tumor cells and euthanasia. A control group of healthy animals was included (n = 6). After euthanasia, macroscopic metastases were assessed and samples of blood were collected. To detect apoptotic bodies in the sera, each sample was mixed with FITC-conjugated annexin V antibody in combination with propidium iodide and then analyzed by flow cytometry. Detection of apoptotic bodies was only significantly increased in the sera of a few tumor bearing animals in late stages of tumor development. Thus, such particles appear not to be the vehicle of the cell-free tumor nucleic acids that are detected at early stages of cancer. PMID- 17108208 TI - Plasma RNA integrity analysis: methodology and validation. AB - The detection of cell-free RNA in plasma and serum of human subjects has found increasing applications in the field of medical diagnostics. However, many questions regarding the biology of circulating RNA remain to be addressed. One issue concerns the molecular nature of these circulating RNA species. We have recently developed a simple and quantitative method to investigate the integrity of plasma RNA. Our results have suggested that cell-free RNA in plasma is generally present as fragmented molecules instead of intact transcripts, with a predominance of 5' fragments. In this article, we summarize the basic principles in the experimental design for plasma RNA integrity analysis and highlight some of the important technical considerations for this type of investigation. PMID- 17108209 TI - Molecular diagnostic markers for lung cancer in sputum and plasma. AB - Lung cancer is the leading cause of cancer deaths worldwide. This study was designed to select multiple DNA markers, which have high sensitivity and specificity to serve as biomarkers for diagnosis of lung cancer. We examined the promoter hypermethylation of three tumor suppressor genes by methylation-specific PCR (MSP), and the instability of eight microsatellite markers by loss of heterozygosity (LOH) and microsatellite instability (MSI) analyses in lung tumor tissues and matched sputum specimens from 79 lung cancer patients. On the basis of the results of sensitivity, specificity, and concordance from each marker analyzed, we selected seven biomarkers, which are LOH of D9S286, D9S942, GATA49D12, and D13S170, MSI of D9S942, and methylation of p16(INK4a) and RARbeta, from the sputum analyses. These selected etiologically associated biomarkers can potentially be used as supplemental diagnostic biomarkers for early lung cancer detection. PMID- 17108210 TI - Quantitative analysis of plasma DNA in colorectal cancer patients: a novel prognostic tool. AB - Extracellular DNA in the plasma or serum of cancer patients has been recently proposed as a source of analyzable cancer-related gene sequences (qualitative approach). Furthermore, patients with different tumor types show high levels of cell-free circulating DNA both in plasma and serum (quantitative approach) at the time of surgery. Our aim was to verify whether the level of cell-free DNA in plasma might help in detecting recurrences during follow-up of colorectal cancer (CRC) patients. We studied 70 patients undergoing surgery for primary CRC. Plasma samples were obtained at the time of surgery and during follow-up. The cell-free circulating DNA in plasma was quantified by the Dipstick Kit method. At the time of surgery, in all patients, cell-free DNA levels in plasma were about 25 times higher in comparison with 20 healthy donors. In contrast, the carcinoembryonic antigen (CEA) value of this cohort of patients was altered in only about 37% of cases. During follow-up, cell-free DNA levels decreased progressively in tumor free patients, while it increased in those developing recurrences or metastases. The results were further supported by qualitative analysis of circulating tumor specific DNA, such as K-Ras mutations and p16(INK4a) promoter hypermethylation. These preliminary data confirm that plasma tumor DNA levels (i) are significantly higher in patients with CRC, (ii) decrease progressively in the follow-up period in tumor-free patients, and (iii) increase in patients with recurrence or metastasis. We suggest, therefore, that the quantification of plasma cell-free DNA might represent a useful tool for monitoring of CRC and, prospectively, for identifying high-risk individuals. PMID- 17108211 TI - Circulating DNA and DNase activity in human blood. AB - The concentration of circulating DNA (cirDNA) and deoxyribonuclease activity in blood plasma of healthy donors and patients with colon or stomach cancer were analyzed. The concentration of DNA was measured using Hoechst 33258 fluorescent assay after the isolation by the glass-milk protocol. A 1-kbp PCR product labeled with biotinylated forward and fluorescein-labeled reverse primers was used as a substrate for DNase. DNase activity was estimated from the data of immunochemical detection of the nonhydrolyzed amplicon. The average concentration of cirDNA in the plasma of healthy donors was low (34 +/- 34 ng/mL), and was accompanied with high DNase activity (0.356 +/- 0.410 U/mL). The increased concentrations of cirDNA in blood plasma of patients with colon and stomach cancer were accompanied by a decrease in DNase activity below the detection level of the assay. The data obtained demonstrate that low DNase activity in blood plasma of cancer patients can cause an increase in the concentration of cirDNA. PMID- 17108212 TI - Comparative analysis of mesenteric and peripheral blood circulating tumor DNA in colorectal cancer patients. AB - An increasing number of reports have demonstrated the presence of tumor-specific DNA in cancer patients' plasma/serum. These findings offer the prospective of serologic tumor markers that may aide in early disease detection, predict subclinical disease progression, and monitor treatment responses. However, for patients with colorectal cancer (CRC), there are few reports using this approach, with most revealing poor sensitivity. In contrast to tumors of other organ systems, CRCs drain predominantly via the mesenteric/portal veins (MV) to the liver. We hypothesize that because of this unique relation, tumor DNA may be less abundant in CRC patients' systemic circulation as compared to the mesenteric/portal system. At the time of surgery, paired blood was collected from both the peripheral vein (PV) and MV from 33 CRC patients. DNA was isolated from serum, quantified and assessed for loss of heterozygosity (LOH) using a panel of 11 microsatellite markers corresponding to regions on six chromosomes frequent for LOH in CRC. In addition, 16 samples were assessed for the presence of hypermethylated DNA for tumor suppressor genes: MGMT, P16, RAR-beta2, RASSF1A, and APC. Circulating tumor DNA associated with LOH or methylation was more frequently detected in the MV of patients, 11 (33%) and 6 (38%), as compared to PV, 9 (27%) and 1 (6%), respectively. This study is the first to identify the presence of increased tumor DNA in the direct efferent venous drainage system of CRC and its variation as compared to systemic circulation. The findings provide important evidence supporting the origin of tumor-associated DNA in circulation, which merits consideration when devising blood-based nucleic acid assays for the assessment of CRC. PMID- 17108213 TI - Real-time quantification of human telomerase reverse transcriptase mRNA in the plasma of patients with prostate cancer. AB - The aim of this study was to evaluate the potential diagnostic value of quantitative analysis of human telomerase reverse transcriptase (hTERT) mRNA in plasma for noninvasive diagnosis of prostate cancer (PCa). Expression levels of hTERT were analyzed by real-time quantitative RT-PCR in 68 patients showing elevated prostate-specific antigen (PSA) levels and a control group of 44 healthy volunteers. Sensitivity and specificity were determined and compared to the corresponding PSA values. Median values for hTERT gene expression in the PCa patients (0.72 ng; range 0.01-12.86) were statistically significantly higher (P < 0.001) than in the control group (0.13 ng; 0.02-0.35). Patients with clinically confirmed prostatitis showed lower plasma hTERT expression than PCa patients (0.29; 0.01-66.07). At a cutoff value of 0.35 sensitivity and specificity for the diagnosis of PCa were 81% and 60%, respectively. We suggest that hTERT mRNA in plasma is a very specific and sensitive method that may aid to differentiate between malignant and nonmalignant prostate tissue and may be a useful marker (in combination with PSA) for early PCa diagnosis. PMID- 17108214 TI - Epigenetic analysis of body fluids and tumor tissues: application of a comprehensive molecular assessment for early-stage breast cancer patients. AB - Breast cancer recurrence is a result of undetected metastasis present at the time of primary patient treatment. More sensitive methods are needed to identify subclinical disease progression to better accompany those increasing advances in early breast cancer screening. Aberrant hypermethylation of tumor-suppressor genes is found frequently in primary breast tumors and has been implicated in disease initiation and progression. Epigenetic characterization of tumor cells may provide highly specific and sensitive molecular surrogates for surveillance. We evaluated whether tumor-associated methylated DNA markers could be identified circulating in bone marrow (BM) aspirates and paired serum samples from 33 early stage patients undergoing surgery for breast cancer. Quantitative methylation specific PCR (qMSP) was performed using a selected tumor-related gene panel for RAR-ss2, MGMT, RASSF1A, and APC. Tumor-associated hypermethylated DNA was identified in 7 (21%) of 33 BM aspirates and 9 (27%) serum samples. In three patients both BM and serum were positive for hypermethylation. The most frequently detected hypermethylation marker was RASSF1A occurring in 7 (21%) patients. Concordance was present between gene hypermethylation detected in BM or serum samples, and matched-pair primary tumors. Advanced AJCC stage was associated with an increased incidence of circulating gene hypermethylation. In addition, methylation patterns in the sentinel lymph node (SLN) metastasis corresponded with that of the primary tumor, confirming epigenetic clonality is associated with early tumor dissemination. This study demonstrates the novel finding of tumor-associated epigenetic markers in BM aspirates/blood and their potential role as targets for molecular detection. PMID- 17108215 TI - Comparison of genetic alterations detected in circulating microsatellite DNA in blood plasma samples of patients with prostate cancer and benign prostatic hyperplasia. AB - Prostate cancer is the most frequent malignant disease and the second most frequent cause of death due to cancer in men in the Western world. Since serum prostate-specific antigen (PSA) and its subforms show poor specificity in clinical practice, a molecular marker for the detection and discrimination of prostate cancer (PCa) could be of great interest. To investigate the potential significance of genetic aberrations, such as loss of heterozygosity (LOH), in PCa we identified and characterized allelic losses in circulating tumor-associated DNA in blood from patients with localized PCa. Genomic DNA extracted from cell free plasma of blood samples drawn from 65 PCa patients was analyzed using a panel of 15 polymorphic microsatellite markers mapping to known tumor-suppressor genes. Comparative analyses were performed with a control group of 36 patients with benign prostatic hyperplasia (BPH). In the current study, we demonstrate that PCa patients had higher DNA concentrations in their blood circulation than BPH patients. In the marker panel studied, LOH was more frequently detected in PCa patients (34%) than in BPH patients (22%). The incidence of LOH in the plasma DNA of PCa patients was highest at chromosomal regions 3p24 (THRB, 22%) and 8p21 (D8S360, 22%) in comparison to the BPH control cohort, which frequently showed LOH at loci 8q21, 8p21, 9p21, and 11q22 (D8S286, D8S360, D9S1748, and D11S898, each 6%). These results indicate that microsatellite analysis using plasma DNA may be an interesting tool for molecular screening of PCa patients. PMID- 17108216 TI - Quantification of total plasma cell-free DNA in ovarian cancer using real-time PCR. AB - Our objective was to compare the levels of total circulating plasma cell-free DNA (CfDNA) using real-time PCR in patients with late-stage ovarian cancer with those in unaffected controls. Following IRB consent, DNA was extracted from archived frozen plasma of 19 patients with primary ovarian carcinoma and 12 age-matched controls using Qiagen DNA Isolation Kits. Quantification of total CfDNA was performed using real-time PCR with the TaqMan Assay for GAPDH, beta-actin and beta-globin and the number of genome equivalents (GE/mL) were determined from a standard curve. CfDNA levels of these loci were compared between the groups with Student's t-test, with P < 0.05 being statistically significant. The mean age of the patients was 61.6 years (+/-9.6) and of the controls was 54 years (+/-12.2). All patients had high-grade, advanced stage (III or IV) serous ovarian carcinomas. Preoperative CA-125 levels ranged from 43 to 15,626 IU/mL (mean 2487.2 +/- 3686 IU/mL). Total CfDNA in ovarian cancer was higher among patients with ovarian cancer as compared to controls at all three loci: GAPDH (P = 0.022), beta-actin (P = 0.025), and beta-globin (P = 0.0089). CfDNA is elevated in advanced stage disease compared to controls. These preliminary results suggest that total CfDNA in the plasma of patients with ovarian cancer may be useful for noninvasive screening and disease surveillance. PMID- 17108217 TI - Cell-free DNA and RNA in plasma as a new molecular marker for prostate and breast cancer. AB - In this study, we examined several molecular markers in prostate and breast cancer patients and in normal individuals. The markers tested were: variations in the quantity of plasma DNA, glutathione-S-transferase P1 gene (GSTP1), Ras association domain family 1A (RASSF1A), and ataxia telangiectasia mutated (ATM) methylation status in plasma, carcinoembryonic antigen (CEA) and prostate specific membrane antigen (PSMA) mRNA in peripheral blood mononuclear cells (PBMC) and plasma samples from prostate cancer patients. DNA quantification in plasma was performed using real-time PCR (RT-PCR). We assessed the methylation status of GSTP1 in plasma DNA using methylation-specific PCR (MSP) assay, while the methylation status of RASSF1A and ATM genes was examined by the MethyLight technology. RT-PCR analysis was used for the detection of mRNA, PSMA, and CEA. In 58.3% of newly diagnosed prostate cancer patients and 26.7% of prostate cancer patients under therapy, plasma DNA levels were increased. Additionally, 48.5% of breast cancer patients showed plasma DNA levels above the cutoff limit. GSTP1 Promotor hypermethylation was detectable in 75% of plasma samples obtained from patients with newly diagnosed prostate cancer and in 36.8% of patients under therapy, whereas 26% and 14% of the breast cancer patients tested were positive for RASSF1A and ATM methylation, respectively. The combination of DNA load and promotor methylation status identified 88% of prostate cancer patients and 54% of breast cancer patients. This study shows that free-circulating DNA can be detected in cancer patients compared with disease-free individuals, and suggests a new, noninvasive approach for early detection of cancer. PMID- 17108218 TI - Early and specific prediction of the therapeutic efficacy in non-small cell lung cancer patients by nucleosomal DNA and cytokeratin-19 fragments. AB - Facing an era of promising new antitumor therapies, predictors of therapy response are needed for the individual management of treatment. In sera collected prospectively from 311 patients with advanced non-small cell lung cancer receiving first-line chemotherapy, changes in nucleosomal DNA fragments, cytokeratin-19 fragments (CYFRA 21-1), carcinoembryonic antigen (CEA), neuron specific enolase (NSE), and progastrin-releasing peptide (ProGRP) were investigated and correlated with therapy response. In univariate analysis, high levels, slower and incomplete decline in nucleosomal DNA, CYFRA 21-1, and CEA predicted poor outcome. DNA concentrations at day 8 of the first therapeutic cycle and CYFRA 21-1 before start of the second cycle were identified as best predictive variables. In multivariate analysis, they predicted progression with a specificity of 100% in 29% of the cases earlier than imaging techniques. Thus, nucleosomal DNA and CYFRA 21-1 specifically identify a subgroup of patients with insufficient therapy response at the early treatment phase and showed to be valuable for disease management. PMID- 17108219 TI - Circulating nucleic acids and diabetic complications. AB - Diabetes mellitus is a major health problem across the world. Diabetic retinopathy (DR) and nephropathy are two of the major complications of diabetes. DR is the leading cause of blindness and diabetic nephropathy is the leading cause of end-stage renal failure. We have examined the potential value of circulating nucleic acids in the detection and monitoring of these two complications of diabetes. mRNA for nephrin was significantly higher in all diabetics compared to healthy controls and it was significantly higher in normoalbuminuric patients compared to healthy controls. This may indicate progression to microalbuminuric stage. Circulating rhodopsin mRNA was detectable in healthy subjects and in diabetic patients. It was significantly raised in diabetic patients with retinopathy. Higher rhodopsin mRNA in diabetic patients without retinopathy suggests that some of them may go on to develop it or already have it subclinically. Circulating nucleic acids have the potential to be noninvasive molecular tests for diabetic complications. PMID- 17108220 TI - Circulating nucleic acids and critical illness. AB - This article reviews some of the early work that has been performed to investigate the potential roles of circulating nucleic acids as prediction markers in acute illness and injury. Circulating DNA and RNA concentrations are elevated early in patients with trauma, stroke and ACS, and are generally highest in patients with a high risk of death. Circulating nucleic acids may be useful markers for the evaluation and risk-stratification of such patients. PMID- 17108221 TI - Cell-free DNA levels as a prognostic marker in acute myocardial infarction. AB - Cell-free DNA that originates from cell death, circulates in peripheral blood. There are indications that the infarcted myocardium contributes to an increase of cell-free DNA levels. Our aims were to quantify levels of cell-free DNA in patients with acute myocardial infarction (AMI) and examine their correlation with myocardial markers and with postinfarction (PI) clinical course. Thirteen patients (age 57 +/- 16 year) admitted with AMI and who underwent thrombolysis with reteplase within 6 h from the onset of chest pain were studied. PB samples were collected on admission and for 5 consecutive days. Creatine kinase (CK) and troponin I (TnI) were measured on admission and every 8 h for 3 consecutive days. Clinical events were recorded throughout the hospitalization period. Cell-free DNA levels were also measured in 30 healthy controls. Log-transformed mean (+/ SE) of maximum free DNA values in patients higher than controls (6873 +/- 357 g.e./mL verses 4112 +/- 234 g.e./mL, P < 0.0001). Log-transformed maximum values of CK and TnI were correlated with log-transformed free DNA values of first (r = 0.62, P = 0.02/r = 0.68, P = 0.01) and second (r = 0.57, P = 0.04/r = 0.72, P = 0.0053) PI day. Nine patients (group A) had an uncomplicated PI clinical course and four patients (group B) had recorded events (three with angina and one death). Free DNA levels on the second PI day were higher in group B than group A (1298.0 +/- 796.0 g.e./mL verses 244.6 +/- 257.7 g.e./mL, P = 0.003). In conclusion, free DNA levels are significantly higher in patients with AMI than in controls and may play a role in the prognosis of these patients. PMID- 17108222 TI - MALDI-TOF mass spectrometry for quantitative, specific, and sensitive analysis of DNA and RNA. AB - Cell-free fetal DNA and RNA released into the maternal circulation offer new opportunities to study fetal and pregnancy-associated abnormalities. Similarly, tumor cells can release cell-free DNA and RNA into the peripheral circulation, and these cell-free DNA and RNA can be used for cancer diagnosis, monitoring, and prognosis. However, these DNA and RNA often exist at very low concentrations (for fetal DNA, approximately 20 genome-equivalents (G.E.)/mL of plasma in the first trimester). The analysis is further complicated by the predominant amount of blood cell-derived DNA and RNA. MALDI-TOF mass spectrometry can provide quantitative, specific, and sensitive analysis of DNA and RNA, and thus may be a useful technology for the field. PMID- 17108223 TI - Rapid prenatal diagnosis by QF-PCR: evaluation of 30,000 consecutive clinical samples and future applications. AB - Rapid prenatal diagnoses of major chromosome abnormalities can be performed on a large scale using highly polymorphic short tandem repeats (STRs) amplified by the quantitative fluorescent polymerase chain reaction (QF-PCR). The assay was introduced as a preliminary investigation to remove the anxiety of the parents waiting for the results by conventional cytogenetic analysis using amniotic fluid or chorionic cells. However, recent studies, on the basis of the analyses of several thousand samples, have shown that this rapid approach has a very high rate of success and could reduce the need for cytogenetic investigations. Its high efficiency, for example, allows early interruption of affected fetuses without the need of waiting for completion of fetal karyotype. The main advantages of the QF-PCR are its accuracy, speed, automation, and low cost that allows very large number of samples to be analyzed by few operators. Here, we report the results of using QF-PCR in a large series of consecutive clinical cases and discuss the possibility that, in a near future, it may even replace conventional cytogenetic analyses on selected samples. PMID- 17108224 TI - Higher amount of free circulating DNA in serum than in plasma is not mainly caused by contaminated extraneous DNA during separation. AB - Circulating DNA isolated from serum and plasma has been shown to be a useful biomarker in various diseases including cancer. Serum reportedly contains a higher amount of free circulating DNA than it does in plasma. The underlying reason for this is unclear, but important because it may have clinical implications in interpreting results and using the appropriate resource. Twenty four pairs of serum and plasma samples were collected from patients with tumors, and free circulating DNA was quantified by real-time quantitative PCR (qPCR) for the ALU repeats, which had a sensitivity of 0.1 pg/microL of DNA in serum/plasma. The possibility of DNA loss was eliminated because ALU-qPCR does not require DNA purification from serum/plasma. The DNA concentrations of serum and plasma samples were 970 +/- 730 pg/microL and 180 +/- 150 pg/microL (mean +/- SD), respectively. The amount of DNA in paired serum and plasma specimens was positively correlated (R = 0.72 and P = 0.0002). An estimated 8.2% of total DNA in serum was extraneous; the concentration of DNA was 6.1 +/- 3.5 (mean +/- SD) fold higher in serum than in paired plasma after subtraction of it. Contribution of extraneous DNA from cells in blood ruptured during the separation step was minor for explaining the difference between serum and plasma. A possible explanation was unequal distribution of DNA during separation from whole blood. We advocate that serum is a better specimen source for circulating cancer-related DNA as a biomarker. PMID- 17108225 TI - Improvement of methods for the isolation of cell-free fetal DNA from maternal plasma: comparison of a manual and an automated method. AB - The low amount of cell-free fetal DNA present in the maternal circulation poses significant challenges to its use in future diagnostic applications, and ways of increasing the yield of this potential marker extracted from maternal plasma are constantly being explored. In this study, we compared two methods of DNA extraction, a manual and an automated method. Our analysis revealed that although the manual method yielded overall more total cell-free DNA, the automated system yielded higher quantities of cell-free DNA of fetal origin. Furthermore, the DNA isolated using the automated system appeared to be of greater purity than that isolated by the manual method, with fewer inhibitors to downstream real-time PCR reactions. PMID- 17108226 TI - The importance of careful blood processing in isolation of cell-free DNA. AB - In healthy individuals, the source of cell-free plasma DNA is predominantly apoptotic, whereas, increased plasma DNA integrity is seen in cancer patients. Therefore, it is important to carefully isolate absolutely "cell-free" plasma DNA. Plasma DNA from 30 healthy females was analyzed using 4 PCR amplicons of increasing size, comparing standard blood processing with additional centrifugation steps prior to DNA extraction. Cellular DNA contamination, indicated by positive amplicons >300 bp was eliminated only after the extra centrifugation step. This highlights the importance of careful processing in preparation of cell-free plasma DNA as a tool for cancer detection and we recommend the use of a microcentrifuge spin, prior to DNA extraction. PMID- 17108227 TI - Nucleosomal DNA fragments in autoimmune diseases. AB - The inadequate response of immune cells to circulating apoptotic products, such as nucleosomal DNA fragments, is assumed to be a potent stimulus for the production of autoantibodies during the pathogenesis and progression of systemic lupus erythematosus (SLE). Here, we analyzed the levels of circulating nucleosomes, caspases, and C-reactive protein in sera of 244 individuals with various autoimmune diseases (155 with autoimmune hepatic disorders, 25 with ANCA associated vasculitis, and 64 with various connective tissue diseases), and 32 healthy controls. Nucleosomes and caspase activities were significantly elevated in sera of patients with hepatic autoimmune diseases, connective tissue diseases, and particularly in ANCA-associated vasculitis when compared with healthy individuals. Nucleosomes showed a correlation with caspases, and caspases with C reactive protein, but nucleosomes did not correlate with C-reactive protein. Serum levels of the apoptotic products, nucleosomes, and caspases are increased in various autoimmune diseases but may not be solely responsible for antinucleosome antibody production in SLE patients. It remains to be clarified whether qualitative changes in nucleosomes are linked with pathogenesis and disease progression in SLE. PMID- 17108228 TI - Concentrations of circulating RNA from healthy donors and cancer patients estimated by different methods. AB - Circulating RNA (cirRNA) was isolated from plasma and cell surface-bound fractions of blood of healthy women and breast cancer patients. RNA samples were DNase treated and quantified by a SYBR Green II assay. Concentrations of RNA sequences of GAPDH, Ki-67 mRNA, and 18S rRNA were measured by real-time quantitative PCR (RT-qPCR) after reverse transcription with random hexamer primers. The obtained data spread over three orders of magnitude for GAPDH and Ki 67 mRNA signals and two orders of magnitude for the copy number of 18S rRNA in blood fractions in both groups. In blood of healthy donors, no correlation was found between the copy number of GAPDH, Ki-67 mRNA, and 18S rRNA and RNA concentrations measured by the SYBR Green II assay. Within the group of breast cancer patients, the concentration GAPDH and Ki-67 mRNA correlated with the concentration of total RNA only in the cell surface-bound fraction; whereas the concentration of 18S rRNA correlated with total RNA in both, the cell surface bound fraction and blood. The copy number of Ki-67 mRNA correlated with copy numbers of GAPDH mRNA in all fractions of cirRNA of healthy donors and breast cancer patients. A correlation between copy numbers of Ki-67 mRNA and 18S rRNA was found only in cell surface-bound fraction of breast cancer patients. The data described here demonstrate the necessity of searching for more suitable RNA markers in order to estimate total cirRNA concentrations by RT-qPCR, although mRNA of GAPDH could be used for normalization of the level of cancer-specific mRNA among patients. PMID- 17108229 TI - Isolation and comparative study of cell-free nucleic acids from human urine. AB - Cell-free nucleic acids (NA) from human urine were investigated. Concentrations of DNA and RNA in the urine of healthy people were independent of gender and were in the range of 6 ng/mL to 50 ng/mL and 24 ng/mL to 140 ng/mL, respectively. DNA fragments of 150-400 bp represent the main part of cell-free DNA, along with DNA fragments up to 1,300 bp, which were found in male urine, and DNA fragments up to 19 kbp, which were found in female urine. Analysis of circulating DNA, isolated from blood of breast cancer patients and cell-free DNA isolated from their urine by methylation-specific PCR, demonstrates that the presence of methylated promoters of RASSF1A and RARbeta2 genes in plasma was accompanied by the detection of the same methylated markers in urine. The data obtained demonstrate applicability of cell-free urine DNA in cancer diagnostics. PMID- 17108230 TI - Influence of mycoplasma contamination on the concentration and composition of extracellular RNA. AB - Kinetics of extracellular RNA accumulation in culture medium and at the cell surface along with their composition and distribution among cell-free and cell surface-bound fractions were investigated in mycoplasma-contaminated and mycoplasma-free HeLa cells. It was shown that the mycoplasma infection influenced the concentration and kinetics of accumulation of total extracellular RNA and the distribution of specific RNA fragments among cell-free and cell-surface-bound fractions. Fragments of immature rRNA were found in culture of mycoplasma infected HeLa cells. The data obtained indicate the existence of selective mechanisms providing binding of RNA with cell surface and their excretion out of cells. PMID- 17108231 TI - Optimized real-time quantitative PCR measurement of male fetal DNA in maternal plasma. AB - DNA of fetal origin is present in the plasma of pregnant women. The quantitative measurement of circulatory fetal DNA (cfDNA) by real-time quantitative PCR (qPCR) has been applied to investigate a possible correlation between increased levels and pregnancy-related disorders. However, as the levels of cfDNA are close to the detection limit (LOD) of the method used, the measurements may not be reliable. This is also problematic for the evaluation of preanalytical steps, such as DNA extraction and cfDNA enrichment by size separation. We optimized a protocol for the qPCR analysis of the multi-copy sequence DYS14 on the Y chromosome. This was compared with an established assay for the single-copy SRY gene. Probit regression analysis showed that the limit of detection (LOD) of the DYS14 assay, (0.4 genome equivalents (GE)) and limit of quantification (LOQ) were 10-fold lower in comparison to SRY (4 GE). The levels of cfDNA obtained from the first trimester of pregnancy could be quantified with high precision by the DYS14 assay (CV below 25%) as opposed to the SRY measurements (26-140%). Additionally, fetal sex was correctly determined in all instances. The low copy numbers of fetal DNA in plasma of women in the first trimester of pregnancy can be measured reliably, targeting the DYS14 that is present in multiple copies per Y chromosome. PMID- 17108232 TI - Theme issue on phenomenology and psychiatry for the 21st century. Taking phenomenology seriously. PMID- 17108233 TI - Activation of human dendritic cells by p-phenylenediamine. AB - Exposure to p-phenylenediamine (pPD), a primary intermediate in hair dye formulations, is often associated with the development of allergic contact dermatitis. Such reactions involve activation of the subject's immune system. The aim of these studies was to explore the relationship between pPD oxidation and functional maturation of human monocyte-derived dendritic cells in vitro. Dendritic cells were incubated with pPD and Bandrowski's base (BB) for 16 h, and expression of the costimulatory receptors CD40, CD80, CD83, CD86, and major histocompatibility complex class II intracellular glutathione levels and cell viability were measured. In certain experiments, glutathione (1 mM) was added to culture medium. Liquid chromatography-mass spectrometry (LC-MS) analysis and exhaustive solvent extraction were used to monitor the rate of [(14)C]pPD oxidation and the extent of pPD binding to cellular and serum protein, respectively. Proliferation of allogeneic lymphocytes was determined by incorporation of [(3)H]thymidine. Exposure of dendritic cells to pPD (5-50 microM), but not BB, was associated with an increase in CD40 and MHC class II expression and proliferation of allogeneic lymphocytes. Dendritic cell activation with pPD was not associated with apoptotic or necrotic cell death or depletion of glutathione. Neither pPD nor BB altered dendritic cell expression of CD80, CD83, or CD86. LC-MS analysis revealed pPD was rapidly oxidized in cell culture media to BB. Addition of glutathione inhibited BB formation but did not prevent covalent binding of pPD to dendritic cell protein or dendritic cell activation. Collectively, these studies show that pPD, but not BB, selectively activates human dendritic cells in vitro. PMID- 17108234 TI - Role of pituitary hormones on 17alpha-ethinylestradiol-induced cholestasis in rat. AB - Estrogens cause intrahepatic cholestasis in susceptible women during pregnancy, after administration of oral contraceptives, or during postmenopausal hormone replacement therapy. 17alpha-Ethinylestradiol (EE) is a synthetic estrogen widely used to cause experimental cholestasis in rodents with the aim of examining molecular mechanisms involved in this disease. EE actions on the liver are thought to be mediated by estrogen receptor alpha (ERalpha) and pituitary hormones. We tested this hypothesis by analyzing metabolic changes induced by EE in livers from hypophysectomized (HYPOX) and hypothyroid rats. Microarray studies revealed that the number of genes regulated by EE was increased almost 4-fold in HYPOX rat livers compared with intact males. Little overlap was apparent between the effects of EE in intact and HYPOX rats, demonstrating that pituitary hormones play a critical role in the hepatic effects of EE. Consistently, hypophysectomy protects the liver against induction by EE of serum bilirubin and alkaline phosphatase, two markers of cholestasis and hepatotoxicity and modulates the effects of EE on several genes involved in bile acid homeostasis (e.g., FXR, SHP, BSEP, and Cyp8b1). Finally, we demonstrate a novel mechanism of action of EE through binding and negative regulation of glucocorticoid receptor-mediated transcription. In summary, pituitary- and ERalpha-independent mechanisms contribute to development of EE-induced changes in liver transcriptome. Such mechanisms may be relevant when this model of EE-induced cholestasis is evaluated. The observation that the pharmacological effects of estrogen in liver differ in the absence or presence of the pituitary could be clinically relevant, because different drugs that block actions of pituitary hormones are now available. PMID- 17108235 TI - Regulation of lipopolysaccharide-induced inducible nitric-oxide synthase expression through the nuclear factor-kappaB pathway and interferon-beta/tyrosine kinase 2/Janus tyrosine kinase 2-signal transducer and activator of transcription 1 signaling cascades by 2-naphthylethyl-6,7-dihydroxy-1,2,3,4 tetrahydroisoquinoline (THI 53), a new synthetic isoquinoline alkaloid. AB - The effects of 2-naphthylethyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline (THI 53), on nitric oxide (NO) production and inducible nitric-oxide synthase (iNOS) protein induction by lipopolysaccharide (LPS) were investigated in RAW 264.7 cells and mice. In cells, THI 53 concentration dependently reduced NO production and iNOS protein induction by LPS. In addition, THI 53 inhibited NO production and iNOS protein induction in LPS-treated mice. LPS-mediated iNOS protein induction was inhibited significantly by the specific tyrosine kinase inhibitor alpha-cyano-(3-hydroxy-4-nitro)cinnamonitrile (AG126) as well as by THI 53. In addition, a c-Jun NH(2)-terminal kinase (JNK) inhibitor anthra[1,9-cd]pyrazole-6 (2H)-one) (SP600125) but not an extracellular regulated kinase inhibitor [2-(2 amino-3-methoxyphenyl)-4H-1-benzopyran-4-one (PD98029)] or a p38 inhibitor [4-(4 fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole (SB230580)] reduced the iNOS protein level induced by LPS. Moreover, a Janus kinase 2 (JAK2) inhibitor alpha-cyano-(3,4-dihydroxy)-N-benzylcinnamide (AG490) dose-dependently prevented LPS-mediated iNOS protein induction. LPS activated phosphorylations of tyrosine kinases, especially tyrosine kinase 2 (Tyk2) and signal transducer and activator of transcription-1 (STAT-1); these were reduced by THI 53. LPS also phosphorylated the JNK pathway; however, this phosphorylation was unaffected by THI 53. Interestingly, a JNK inhibitor (SP600125) and another tyrosine kinase inhibitor (genistein) significantly inhibited STAT-1 phosphorylation, suggesting that the LPS-activated JNK pathway and a tyrosine kinase pathway (especially Tyk2) may link to the STAT-1 pathway, which is involved in iNOS induction. However, THI 53 regulates LPS-mediated iNOS protein induction by affecting the Tyk2/JAK2-STAT-1 pathway, not the JNK pathway. The inhibition by THI 53 of LPS induced NO production was recovered by a tyrosine phosphatase inhibitor (Na(3)VO(4)), which supports the possibility that THI 53 inhibits the LPS-induced inflammatory response through regulation of tyrosine kinase pathways. THI 53 also inhibited LPS-mediated interferon (IFN)-beta production and nuclear factor-kappaB (NF-kappaB) activation. Thus, THI 53 may regulate LPS-mediated inflammatory response through both the NF-kappaB and IFN-beta/Tyk2/JAK2-STAT-1 pathways. PMID- 17108236 TI - A noninflammatory interleukin-1beta fragment stimulates fetal lung fluid absorption in guinea pigs. AB - We have previously demonstrated that full-length interleukin (IL)-1beta can induce and stimulate lung fluid absorption in near-term guinea pig fetuses via stimulation of fetal cortisol synthesis and release. To develop a potentially clinically useful drug, we tested the hypothesis that maternal administration of a noninflammatory IL-1beta-fragment (IL-1beta(Fr)) induced cortisol synthesis and stimulated lung fluid absorption in preterm fetuses. IL-1beta(Fr) was administered s.c. daily to timed-pregnant guinea pigs for 3 days with and without simultaneous cortisol synthesis inhibition by metyrapone. Fetuses were obtained by abdominal hysterotomy at 61 and 68 days gestation and instilled with isosmolar 5% albumin into the lungs, and lung fluid absorption was measured over 1 h by mass balance. Lung fluid absorption was induced at 61 days and stimulated at 68 days gestation by IL-1beta(Fr), which both were attenuated by cortisol synthesis inhibition. Moreover, induction of labor by oxytocin stimulated lung fluid absorption at 61 days but had no stimulatory effect at 68 days gestation when given with the IL-1beta(Fr). Plasma adrenocorticotropin and cortisol concentrations were increased by IL-1beta(Fr) at 61 days gestation and remained high but unstimulated by IL-1beta(Fr) at 68 days gestation, and metyrapone always reduced cortisol concentrations. Prenatal lung fluid absorption, when present as well as IL-1beta(Fr)-induced, was always propranolol- and amiloride-sensitive, suggesting that beta-adrenoceptor stimulation and the epithelial Na(+) channel (ENaC) were critical for the induced/stimulated lung fluid absorption. ENaC expression was increased by IL-1beta(Fr) and attenuated by cortisol synthesis inhibition. Thus, our results suggest a potential clinical use of IL-1beta(Fr) therapeutically to induce lung fluid absorption in fetuses at risk of preterm delivery. PMID- 17108237 TI - Human multidrug resistance protein 2 transports the therapeutic bile salt tauroursodeoxycholate. AB - The multidrug resistance protein 2 (MRP2/ABCC2) mediates the biliary excretion of glucuronide and glutathione conjugates of endogenous and exogenous compounds. We examined the activation of human MRP2-mediated ATP-dependent transport by the choleretic bile salt ursodeoxycholic acid (UDC) and its taurine and glycine amidates in Sf9 cell membranes expressing MRP2 using beta-estradiol 17-(beta-D glucuronide) (E(2)17G) and beta-estradiol 3-(beta-D-glucuronide) (E(2)3G) as substrates. MRP2 transported E(2)3G via classic Michaelis-Menten kinetics (K(m) = 122 microM; V(max) = 3.0 nmol/mg/min), whereas E(2)17G transport showed positive cooperativity (Hill slope, 2.15; K(m) = 75 microM; V(max) = 3.8 nmol/mg/min). UDC, tauroursodeoxycholate, and glycoursodeoxycholate (80-100 microM) maximally stimulated E(2)3G transport 9-, 7.9-, and 3.6-fold, respectively, whereas higher concentrations (1-2 mM) inhibited transport. At low (0.3 microM) concentrations, tauroursodeoxycholate was transported only in the presence of E(2)17G or E(2)3G, but not other MRP2 substrates such as methotrexate, leukotriene C(4), or S methylglutathione. Kinetic analysis of higher concentrations of tauroursodeoxycholate transport by MRP2 showed positive cooperativity (Hill slope, 1.84; K(m) = 127 microM; V(max) = 779 pmol/mg/min). Taurocholate (2-100 microM) was not detectably transported by MRP2 either alone or in the presence of E(2)17G but was transported in the presence of E(2)3G. Thus, UDC, tauroursodeoxycholate, and glycoursodeoxycholate activated MRP2 transport. Tauroursodeoxycholate was transported by MRP2 and demonstrated positive cooperativity, identifying it as the second MRP2 substrate able to stimulate its own transport. The data suggest MRP2 binding sites that can require specific complementarities between substrates and modulators of MRP2-mediated transport. PMID- 17108239 TI - Teaching and learning in the interactive classroom. AB - The Claude Bernard Distinguished Lectureship of the Teaching of Physiology Section is presented annually at the Experimental Biology meeting. The lectureship is named for Prof. Claude Bernard, the experimental physiologist who is credited with introducing the concept of homeostasis. The 2006 Claude Bernard Distinguished Lecture was given by Dr. Dee U. Silverthorn from the University of Texas at Austin, TX. PMID- 17108240 TI - Teaching aldosterone regulation and basic scientific principles using a classic paper by Dr. James O. Davis and colleagues. AB - Classroom discussion of scientific articles can be an effective means of teaching scientific principles and methodology to both undergraduate and graduate science students. The availability of classic papers from the American Physiological Society Legacy Project has made it possible to access articles dating back to the early portions of the 20th century. In this article, we discuss a classic paper from the laboratory of Dr. James O. Davis on the regulation of aldosterone synthesis from the adrenal zona glomerulosa cell. Dr. Davis has conducted much of the seminal research investigating the renin-angiotensin system and the regulation of aldosterone release by angiotensin II. In addition to a characterization of the effects of ACTH on aldosterone regulation, this study is useful for discussing the basic principles of negative feedback pathways of the hypothalamic-pituitary axis. This study also provides examples of early bioassay techniques for the detection of angiotensin II and of the importance of quantitative measurements when investigating physiological responses. Three figures and one table are reproduced from the original article along with a series of discussion questions designed to facilitate discovery learning. PMID- 17108241 TI - PGC-1alpha: a key regulator of energy metabolism. AB - Peroxisome proliferator-activated receptor-gamma coactivator (PGC)-1alpha is a member of a family of transcription coactivators that plays a central role in the regulation of cellular energy metabolism. It is strongly induced by cold exposure, linking this environmental stimulus to adaptive thermogenesis. PGC 1alpha stimulates mitochondrial biogenesis and promotes the remodeling of muscle tissue to a fiber-type composition that is metabolically more oxidative and less glycolytic in nature, and it participates in the regulation of both carbohydrate and lipid metabolism. It is highly likely that PGC-1alpha is intimately involved in disorders such as obesity, diabetes, and cardiomyopathy. In particular, its regulatory function in lipid metabolism makes it an inviting target for pharmacological intervention in the treatment of obesity and Type 2 diabetes. PMID- 17108242 TI - A role for transcription factor NF-kappaB in autoimmunity: possible interactions of genes, sex, and the immune response. AB - Sex hormones have long been implicated in autoimmune diseases because women account for 80% of cases. The mechanism of hormonal action in autoimmunity is unknown. Drawing on genetic studies of autoimmune disease, this article discusses how both genes and sex hormones may exert their effects through the same general mechanism, dysregulation of transcription factor NF-kappaB, an immunoregulatory protein. Gene and hormone alterations of the NF-kappaB signaling cascade provide a unifying hypothesis to explain the wide-ranging human and murine autoimmune disease phenotypes regulated by NF-kappaB, including cytokine balance, antigen presentation, lymphoid development, and lymphoid repertoire selection by apoptosis. PMID- 17108243 TI - Where's the evidence that active learning works? AB - Calls for reforms in the ways we teach science at all levels, and in all disciplines, are wide spread. The effectiveness of the changes being called for, employment of student-centered, active learning pedagogy, is now well supported by evidence. The relevant data have come from a number of different disciplines that include the learning sciences, cognitive psychology, and educational psychology. There is a growing body of research within specific scientific teaching communities that supports and validates the new approaches to teaching that have been adopted. These data are reviewed, and their applicability to physiology education is discussed. Some of the inherent limitations of research about teaching and learning are also discussed. PMID- 17108244 TI - Peer instruction enhanced student performance on qualitative problem-solving questions. AB - We tested the hypothesis that peer instruction enhances student performance on qualitative problem-solving questions. To test this hypothesis, qualitative problems were included in a peer instruction format during our Physiology course. Each class of 90 min was divided into four to six short segments of 15 to 20 min each. Each short segment was followed by a qualitative problem-solving scenario that could be answered with a multiple-choice quiz. All students were allowed 1 min to think and to record their answers. Subsequently, students were allowed 1 min to discuss their answers with classmates. Students were then allowed to change their first answer if desired, and both answers were recorded. Finally, the instructor and students discussed the answer. Peer instruction significantly improved student performance on qualitative problem-solving questions (59.3 +/- 0.5% vs. 80.3 +/- 0.4%). Furthermore, after peer instruction, only 6.5% of the students changed their correct response to an incorrect response; however, 56.8% of students changed their incorrect response to a correct response. Therefore, students with incorrect responses changed their answers more often than students with correct responses. In conclusion, pausing four to six times during a 90-min class to allow peer instruction enhanced student performance on qualitative problem-solving questions. PMID- 17108245 TI - Staff and student perceptions of computer-assisted assessment for physiology practical classes. AB - Effective assessment of laboratory practicals is a challenge for large-size classes. To reduce the administrative burden of staff members without compromising the student learning experience, we utilized dedicated computer software for short-answer question assessment for nearly 300 students and compared it with the more traditional, paper-based method of assessment of the same student cohort. Students were generally favorably disposed toward computer assisted assessment (CAA): 75% of the students responded that for future assignments, they either had no preference for the method of assessment or would prefer CAA. Advantages were perceived to be remote access to the questions and ease of submission. The most common disadvantage cited was lack of internet access. Various advantages of CAA were mentioned by staff members: notably, the reduction in marking time and reduction of paperwork as well as the potential for the software to detect plagiarism and to administer anonymous marking. Disadvantages to CAA were the need to tailor questions to the technology, having to adapt to reading answers and marking onscreen, and the quality of feedback to students. All of the disadvantages could be overcome by training and improved versions of CAA software, currently under development. The use of CAA has proved to be a welcome addition to the tools available to staff members for the assessment of practical classes, and future improved versions of the software will increase the utility of this assessment method. PMID- 17108246 TI - Promoting self-directed learning using a menu of assessment options: the investment model. AB - Undergraduate science students took an Inquiry course in their second (sophomore) year. The course was designed to explore the social life of scientific knowledge. They were given a set of eight assessment options: personal logs, targeted oral examinations, commentaries, mini-lectures, individual explorations, research proposals, book reviews, and problem-solving exercises. Each option had a specific maximum mark (percentage or grade point) associated with it. Students were permitted to select any set of options to obtain their total grade for the course. From the student's perspective, the course provided a valuable learning experience and enabled them to recognize the complexities involved in the process of generating scientific information and making it useful and relevant to the public. The opportunity given to select their own assessment options enhanced their learning. For me, as the sole instructor managing 51 students, the experience was rewarding. PMID- 17108247 TI - Elementary education majors experience hands-on learning in introductory biology. AB - Faculty members from the University of South Dakota attended the Curriculum Reform Institute offered by the University of Wisconsin at Oshkosh, WI, during the summer of 2002 to design a course sequence for elementary education majors that better meets their needs for both content and pedagogy based on the science education standards. The special section of introductory biology that resulted from this workshop is designed to use laboratories and activities that either help students learn major concepts in the life sciences or model how to teach these concepts to their future K-8 students. This study describes how the active, hands-on learning opportunity for preservice teachers with its emphasis on both content and performance-based assessment was implemented in an introductory biology course for elementary education majors during the spring of 2004. During the initial offering of this course, student perceptions about what helped them to learn in the special section was compared with their nonscience major peers in the large lecture-intensive class that they would have taken. Each group of students completed early and late web-based surveys to assess their perceptions about learning during the courses. After the completion of the course, students in the special section appreciated how the relevance of science and conducting their own scientific experimentation helped them learn, enjoyed working and studying in small groups, valued diverse class time with very little lecture, were more confident in their abilities in science, and were more interested in discussing science with others. This course format is recommended for science classes for preservice teachers. PMID- 17108248 TI - It's difficult to change the way we teach: lessons from the Integrative Themes in Physiology curriculum module project. AB - The Integrative Themes in Physiology (ITIP) project was a National Science Foundation-funded collaboration between the American Physiological Society (APS) and the Human Anatomy and Physiology Society (HAPS). The project goal was to create instructional resources that emphasized active learning in undergraduate anatomy and physiology classrooms. The resources (activity modules and professional development) addressed two factors thought to be limiting science education reform: instructors' knowledge of how to implement active learning instruction and time to design innovative curricula. Volunteer instructors with a strong interest in using active learning in their classrooms were recruited to use the ITIP modules and provide ease-of-use feedback and student assessment data. As the study unfolded, instructor attrition was higher than had been anticipated, with 17 of 36 instructors withdrawing. More surprisingly, instructors remaining with the project failed to use the modules and reported specific obstacles that precluded module use, including lack of support from academic leadership, unplanned class size increases and heavy teaching loads, a union strike, insufficient time to develop a mindset for change, inadequate technology/funding, an adverse human subjects ruling, incompatibility of modules with instructors' established content and expectations, and personal factors. Despite the lack of module use and obstacles, 8 of 19 site testers began independently to introduce new active learning instruction into their classrooms. In the larger picture, however, it is important to note that only 8 of the initial 36 volunteers (22%) actually ended up changing their instruction to include opportunities for student active learning. These findings underscore the difficulty of implementing instructional change in college classrooms. PMID- 17108249 TI - Case study analysis and the remediation of misconceptions about respiratory physiology. AB - Most students enter the physiology classroom with one or more fundamental misconceptions about respiratory physiology. This study examined the prevalence of four respiratory misconceptions and determined the role of case analysis in the remediation of one of them. A case study was used to help students learn about oxygen transport in the blood and a conceptual diagnostic test was used to assess student understanding of the relation between Po(2) and hemoglobin saturation by probing for the corresponding (Sa/Po(2)) misconception. A 36% remediation of the Sa/Po(2) misconception was found to be associated with case analysis. This repair was selective since the frequency of three other respiratory misconceptions was found to be unchanged after classroom instruction about respiratory physiology in lectures and laboratories. Remediation of the Sa/Po(2) misconception before an instructor-led, in-class case review was superficial and temporary. Explanations provided by students who correctly answered the Sa/Po(2) conceptual diagnostic test showed improved conceptual understanding following case analysis. These results suggest that a learning strategy where students actively confront their faulty notions about respiratory physiology is useful in helping them overcome their misconceptions. PMID- 17108250 TI - Case-based learning of blood oxygen transport. AB - A case study about carbon monoxide poisoning was used help students gain a greater understanding of the physiology of oxygen transport by the blood. A review of student answers to the case questions showed that students can use the oxygen-hemoglobin dissociation curve to make meaningful determinations of oxygen uptake and delivery. However, the fact that many students had difficulty locating the effect of carbon monoxide poisoning in the process of external respiration suggests that these students have not built a robust model of how oxygen distributes itself between the plasma and hemoglobin. This suggests that more determined emphasis on how oxygen enters the blood and how it is partitioned between hemoglobin and the plasma would help students develop more accurate mental models of how oxygen moves from the lungs to the tissues. PMID- 17108251 TI - Use of modified SOAP notes and peer-led small-group discussion in a Medical Physiology course: addressing the hidden curriculum. AB - Peer leading of small-group discussion of cases; use of modified subjective, objective, assessment of physiology (SOAP) notes; and opportunities for self assessment were introduced into a Medical Physiology course to increase students' awareness and practice of professional behaviors. These changes arose from faculty members' understanding of the hidden curriculum and their efforts to reveal it to take increased advantage of its educationally beneficial aspects. Faculty members and students observed that the requirement for students to submit SOAP notes before their discussions meant that they were well prepared to participate. Student satisfaction with the protocol was high, with >95% of the students agreeing that discussants were well prepared and that the overall performance of their discussion group was good. A comparison of students' performance on selected exam questions showed that peer leading was equally as effective as a previously used teacher-centered approach. Students agreed that their ability to analyze a clinical case had improved using this protocol, an effect that persisted at least one semester after the end of the course. These approaches were time and cost efficient from a faculty perspective while serving the needs of the students. The use of SOAP notes and peer-led discussion were effective forms of instruction, in which students succeeded in learning medical physiology and in practicing professional behaviors. PMID- 17108252 TI - Isolation and culture of bovine oviductal epithelial cells for use in the anatomy and physiology laboratory and undergraduate research. AB - This article presents methods for the isolation and culture of epithelial cells from the bovine oviduct for use in both research and the teaching laboratory and provides examples of ways that an oviductal cell culture can be incorporated into an undergraduate research program. Cow reproductive tracts are readily available from area butchers, and the procedure for isolation of the epithelium is simple and inexpensive. The cells can be observed immediately after isolation or can be cultured for up to 72 h under simple conditions for observation over several days. For experimental use, epithelial cells are cultured in standard cell culture medium, where they continue to divide and actively secrete substances into the medium. The ease with which the tissue can be collected and cells isolated makes the oviductal epithelium ideal for use in both the teaching laboratory and research projects in which undergraduates serve as investigators. PMID- 17108253 TI - Study of physiological responses to acute carbon monoxide exposure with a human patient simulator. AB - Human patient simulators are widely used to train health professionals and students in a clinical setting, but they also can be used to enhance physiology education in a laboratory setting. Our course incorporates the human patient simulator for experiential learning in which undergraduate university juniors and seniors are instructed to design, conduct, and present (orally and in written form) their project testing physiological adaptation to an extreme environment. This article is a student report on the physiological response to acute carbon monoxide exposure in a simulated healthy adult male and a coal miner and represents how 1) human patient simulators can be used in a nonclinical way for experiential hypothesis testing; 2) students can transition from traditional textbook learning to practical application of their knowledge; and 3) student initiated group investigation drives critical thought. While the course instructors remain available for consultation throughout the project, the relatively unstructured framework of the assignment drives the students to create an experiment independently, troubleshoot problems, and interpret the results. The only stipulation of the project is that the students must generate an experiment that is physiologically realistic and that requires them to search out and incorporate appropriate data from primary scientific literature. In this context, the human patient simulator is a viable educational tool for teaching integrative physiology in a laboratory environment by bridging textual information with experiential investigation. PMID- 17108254 TI - Anthropometric evaluations of body composition of undergraduate students at the University of La Reunion. AB - A positive correlation is well established between obesity and the susceptibility to develop metabolic syndrome, a multifactorial disease dramatically associated with an enhanced mortality risk in the developed world. A high prevalence of obesity has recently been described at La Reunion Island, a French department in the Indian Ocean. Anthropometry is generally considered as the single most easily obtainable, inexpensive, and noninvasive method that reflects body composition. At the University of La Reunion, a laboratory course involving students was designed to teach anthropometric measurements for the determination of body composition. Using skin fold thickness equations, students determined the fat and total muscular masses of their body composition. The influences of sex and their physical activity or inactivity on these different parameters were compared and interpreted at the end of the course. Positive and significant correlations were established between the students' body mass indexes values and their fat mass percentages and between their fat-free and muscular masses as well. A higher fat mass percentage was found in sedentary students compared with more active ones. Therefore, this laboratory makes the students practice and understand the use of classical techniques to evaluate the body composition of a person. It also alerts them to the correlation between a sedentary attitude and higher body fat content. This laboratory course constitutes an active introduction to a following lesson on more recent and actual techniques used to evaluate body composition. PMID- 17108255 TI - Teaching insect retinal physiology with newly designed, inexpensive micromanipulators. AB - In this article, we detail how to produce two inexpensive micromanipulators that offer high precision (approximately 25 microm) along a single axis of movement. The more expensive of the designs provides improved versatility along multiple axes. Both manipulators offer substantial savings over commercially available micromanipulators with comparable capabilities. Plans and instructions are given such that a novice can produce the manipulators with simple tools. The manipulators are designed to serve undergraduate teaching exercises in physiology. An electroretinogram exercise is suggested in adult house flies (Musca) or flesh flies (Neobellieria). Measuring the intensity-response function and temporal characteristics of visual transduction are discussed. A brief introduction to the field of visual transduction and the physiology of the laboratory exercises is provided as well. PMID- 17108257 TI - Translating old units of measurements. PMID- 17108258 TI - A midlecture student seminar: an activity to break the monotony. PMID- 17108259 TI - Starting physiology: understanding homeostasis. PMID- 17108260 TI - Inhibition of tumor necrosis factor-alpha-inducible inflammatory genes by interferon-gamma is associated with altered nuclear factor-kappaB transactivation and enhanced histone deacetylase activity. AB - Airway smooth muscle (ASM) cells can act as effector cells in the initiation and/or perpetuation of airway inflammation in asthma by producing various inflammatory chemokines or cytokines. Previous studies from our laboratory and others showed that the combination of tumor necrosis factor-alpha (TNFalpha) and interferon-gamma (IFNgamma) or endogenous IFNbeta results in a synergistic induction of various pro-inflammatory genes, including CD38 and regulated upon activation normal T-cell expressed and secreted (RANTES), in ASM cells. In contrast to these studies, we found that IFNgamma (1000 U/ml) markedly inhibited TNFalpha-induced expression of interleukin (IL)-6, IL-8, and eotaxin by 66.29+/ 3.33, 43.86+/-7.11, and 63.25+/-6.46%, respectively. These genes were also found to be NF-kappaB-dependent in that TNFalpha-induced expression of IL-6, IL-8, and eotaxin was dose-dependently inhibited by the selective IKKbeta inhibitor 4-(2' aminoethyl)amino-1,8-dimethylimidazo[1,2-a]quinoxaline (BMS-345541) (1-30 microM). Using a luciferase reporter construct containing kappaB sites, we found that IFNgamma (10-1000 U/ml) inhibits NF-kappaB-dependent gene transcription in a dose-dependent manner. Moreover, IFNgamma failed to affect TNFalpha-induced IkappaKbeta phosphorylation or IkappaB degradation as well as nuclear NF kappaB/DNA interaction. It is noteworthy that IFNgamma decreases TNFalpha-induced histone acetyl transferase (HAT) and increases histone deacetylase (HDAC) activities. Finally, trichostatin A, an HDAC inhibitor, prevents IFNgamma inhibitory action on TNFalpha-induced gene expression. Together, our data indicate that IFNgamma is a potent inhibitor of specific TNFalpha-inducible inflammatory genes by acting on NF-kappaB transactivation via the modulation of HDAC function. PMID- 17108261 TI - Agonist-, antagonist-, and benzodiazepine-induced structural changes in the alpha1 Met113-Leu132 region of the GABAA receptor. AB - The structural basis by which agonists, antagonists, and allosteric modulators exert their distinct actions on ligand-gated ion channels is poorly understood. We used the substituted cysteine accessibility method to probe the structure of the GABAA receptor in the presence of ligands that elicit different pharmacological effects. Residues in the alpha1 Met113-Leu132 region of the GABA binding site were individually mutated to cysteine and expressed with wild-type beta2 and gamma2 subunits in Xenopus laevis oocytes. Using electrophysiology, we determined the rates of reaction of N-biotinaminoethyl methaneth-iosulfonate (MTSEA-biotin) with the introduced cysteines in the resting (unliganded) state and compared them with rates determined in the presence of GABA (agonist), 4-[6 imino-3-(4-methoxyphenyl)pyridazin-1-yl]butanoic acid hydrobromide (SR-95531; antagonist), pentobarbital (allosteric modulator), and flurazepam (allosteric modulator). alpha1N115C, alpha1L117C, alpha1T129C, and alpha1R131C are predicted to line the GABA binding pocket because MTSEA-biotin modification of these residues decreased the amount of current elicited by GABA, and the rates/extents of modification were decreased both by GABA and SR-95531. Reaction rates of some substituted cysteines were different depending on the ligand, indicating that barbiturate- and GABA-induced channel gating, antagonist binding, and benzodiazepine modulation induce specific structural rearrangements. Chemical reactivity of alpha1E122C was decreased by either GABA or pentobarbital but was unaltered by SR-95531 binding, whereas alpha1L127C reactivity was decreased by agonist and antagonist binding but not affected by pentobarbital. Furthermore, alpha1E122C, alpha1L127C, and alpha1R131C changed accessibility in response to flurazepam, providing structural evidence that residues in and near the GABA binding site move in response to benzodiazepine modulation. PMID- 17108262 TI - Multidrug-resistant pulmonary infection in cystic fibrosis--what does 'resistant' mean? PMID- 17108263 TI - The diversity of definitions of multidrug-resistant (MDR) and pandrug-resistant (PDR) Acinetobacter baumannii and Pseudomonas aeruginosa. AB - Different definitions of the terms multidrug-resistant (MDR) and pandrug resistant (PDR) Acinetobacter baumannii and Pseudomonas aeruginosa have been used in the biomedical literature. The authors searched for relevant studies indexed in the PubMed database (01/2000-09/2005) to systematically examine the various definitions of MDR and PDR for these bacteria. Initially 107 retrieved relevant studies were reviewed. Ninety-two studies were further analysed, 50 of which focused on A. baumannii and 42 on P. aeruginosa. A considerable diversity of definitions of the terms MDR and PDR A. baumannii and P. aeruginosa was found. Of note, the term PDR was inappropriately used in all five studies that used it. The review reveals that various definitions have been used for the terms MDR and PDR A. baumannii and P. aeruginosa, a fact that causes confusion to researchers and clinicians. The authors believe that at least a widely accepted definition for PDR A. baumannii and P. aeruginosa should be uniformly used worldwide. PMID- 17108264 TI - fliP influences Citrobacter koseri macrophage uptake, cytokine expression and brain abscess formation in the neonatal rat. AB - Citrobacter koseri causes neonatal meningitis frequently complicated with multiple brain abscesses. During C. koseri central nervous system infection in the neonatal rat model, previous studies have documented many bacteria-filled macrophages within the neonatal rat brain and abscesses. Previous studies have also shown that C. koseri is taken up by, survives phagolysosomal fusion and replicates in macrophages in vitro and in vivo. In this study, in order to elucidate genetic and cellular factors contributing to C. koseri persistence, a combinatory technique of differential fluorescence induction and transposon mutagenesis was employed to isolate C. koseri genes induced while inside macrophages. Several banks of mutants were subjected to a series of enrichments to select for gfp : : transposon fusion into genes that are turned off in vitro but expressed when intracellular within macrophages. Further screening identified several mutants attenuated in their recovery from macrophages compared with the wild-type. A mutation within an Escherichia coli fliP homologue caused significant attenuation in uptake and hypervirulence in vivo, resulting in death within 24 h. Furthermore, analysis of the immunoregulatory interleukin (IL)-10/IL 12 cytokine response during infection suggested that C. koseri fliP expression may alter this response. A better understanding of the bacteria-macrophage interaction at the molecular level and its contribution to brain abscess formation will assist in developing preventative and therapeutic strategies. PMID- 17108265 TI - Protease IV production in Pseudomonas aeruginosa from the lungs of adults with cystic fibrosis. AB - Protease IV is important in the pathogenesis of Pseudomonas aeruginosa-induced microbial keratitis, but little is known of its role in cystic fibrosis (CF) lung infection. In this study protease IV production was examined in 43 P. aeruginosa isolates (24 non-clonal and 19 clonal) from the lungs of chronically infected adult patients attending the Royal Prince Alfred Hospital CF Clinic, Sydney, Australia. Overall, 32/43 (74 %) isolates were positive for protease IV protein by Western blotting and 22/43 (51 %) had evidence of active protease IV on gelatin zymography. Clonal strains were 1.6 times more likely than non-clonal strains to produce protease IV [18/19 (95 %) versus 14/24 (58 %), RR=1.6, CI 1.1 2.3, P=0.007] and 3 times more likely to secrete the protein [16/19 (84 %) versus 6/24 (25 %), RR=3.4, CI 1.6-6.9, P<0.001]. Nine of the ten strains negative by both Western blotting and zymography were non-clonal, and all but one of these was positive for the protease IV gene. There was a marked strain-to-strain variation in the amount of protease IV produced. Secretion of protease IV by clonal strains may enhance their infectivity and ability to adapt to the changing CF lung environment. Overall the findings suggest that protease IV plays an important role in the pathogenesis of P. aeruginosa infection in the CF lung. PMID- 17108266 TI - Staphylococcal secretory inhibitor of platelet microbicidal protein is associated with prostatitis source. AB - This study reports the detection of an extracellular staphylococcal product, designated secretory inhibitor of platelet microbicidal protein (SIPMP), that causes local inhibition of the bactericidal action of platelet microbicidal protein (PMP) in the fluid phase. Urethral isolates of Staphylococcus aureus (n=24) and coagulase-negative staphylococci (CNS) (n=47) from patients with or without chronic bacterial prostatitis (CBP) were tested. SIPMP production was tested by inhibition of PMP bioactivity against Bacillus subtilis and was expressed as percentage inhibition of PMP bactericidal activity. The PMP susceptibility of staphylococcal strains was determined by exposing bacterial cells to serial dilutions of PMP. Staphylococci from patients without CBP produced SIPMP at levels of 10.3+/-1.2 and 13.25+/-1.72 % for S. aureus and CNS, respectively. Strains isolated from men with CBP inhibited PMP-induced killing of B. subtilis by 23.38+/-4.2 % (P<0.05) and 23.69+/-1.87 % (P<0.01) for S. aureus and CNS, respectively. SIPMP production correlated with staphylococcal resistance to PMP (r2=0.6082 and 0.7264 for S. aureus and CNS, respectively). SIPMP represents a hitherto unrecognized determinant of staphylococcal pathogenicity. These results suggest that SIPMP production is associated with the CBP source. Data from this study may have significant implications for the understanding of the pathogenesis of CBP. PMID- 17108267 TI - Impaired immune response to Candida albicans in aged mice. AB - The prevalence of opportunistic fungal infections has increased dramatically among the aged population in recent years. This work investigated the effect of ageing on murine defences against Candida albicans. Aged C57BL/6 mice that were experimentally infected intravenously had a significantly impaired survival and a higher tissue fungal burden compared with young mice. In vitro production of tumour necrosis factor (TNF)-alpha by macrophages from aged mice in response to yeast cells and hyphae of C. albicans was significantly lower than production by macrophages from young mice. In vitro production of cytokines, such as TNF-alpha and gamma interferon (IFN-gamma), by antigen-stimulated splenocytes from mice intravenously infected with C. albicans cells was also diminished in old mice. This decrease in production of T helper 1 cytokines in old mice correlated with a diminished frequency of IFN-gamma-producing CD4+ T lymphocytes, although the ability to develop an acquired resistance upon vaccination (primary sublethal infection) of mice with the low-virulence PCA2 strain was not affected in aged mice. The diversity of antigens recognized by C. albicans-specific antibodies in sera from infected aged mice was clearly diminished when compared with that from infected young mice. Taken together, these data show that aged mice develop an altered innate and adaptive immune response to C. albicans and are more susceptible to systemic primary candidiasis. PMID- 17108268 TI - Rapid and low-cost colorimetric method using 2,3,5-triphenyltetrazolium chloride for detection of multidrug-resistant Mycobacterium tuberculosis. AB - A rapid and inexpensive method for the detection of drug resistance in Mycobacterium tuberculosis is essential for the effective control of tuberculosis. The aim of this study was to evaluate a colorimetric method using 2,3,5-triphenyltetrazolium chloride (TTC) for antibiotic susceptibility testing of M. tuberculosis isolates. Eleven multidrug-resistant (MDR) isolates of M. tuberculosis and 12 isolates which were susceptible to rifampicin (RIF) and isoniazid (INH) were used. The test was performed with a critical concentration of 0.2 microg ml(-1) for INH and 2.0 microg ml(-1) for RIF in 7H9GC broth with 0.625 microg TTC ml(-1). Each isolate was inoculated under these conditions and inspected daily for colour changes; the results were obtained after a mean of 4.9 days. The sensitivity and specificity of this method were 100 % and 92 %, respectively, for both antibiotics. Considering the speed, technical ease and cost-effectiveness of this method, the TTC assay is a good alternative method for drug susceptibility testing of M. tuberculosis isolates. PMID- 17108269 TI - Isolation of Salmonella enterica subspecies enterica serovar Paratyphi B dT+, or Salmonella Java, from Indonesia and alteration of the d-tartrate fermentation phenotype by disrupting the ORF STM 3356. AB - Salmonella enterica subspecies enterica serovar Paratyphi B [O1,4,(5),12 : Hb : 1,2] can cause either an enteric fever (paratyphoid fever) or self-limiting gastroenteritis in humans. The d-tartrate non-fermenting variant S. enterica subsp. enterica serovar Paratyphi B dT- (S. Paratyphi B) is the causative agent of paratyphoid fever, and the d-tartrate fermenting variant S. enterica subsp. enterica serovar Paratyphi B dT+ (S. Paratyphi B dT+; formerly called Salmonella Java) causes gastroenteritis. S. Java is currently recognized as an emerging problem worldwide. Twelve dT+ S. Java isolates were collected in Indonesia between 2000 and 2002. One-third of them contained Salmonella genomic island 1 (SGI1), which gives the multidrug-resistant phenotype to the bacteria. In this study, a PCR-based method to detect a single nucleotide difference responsible for the inability to ferment d-tartrate, reported elsewhere, was validated. The d tartrate fermenting phenotype of S. Java was converted to the non-fermenting phenotype by the disruption of the ORF STM 3356, and the d-tartrate non fermenting phenotype of the ORF STM 3356-disrupted strain and the dT- reference strain was changed to the dT+ phenotype by complementing ORF STM 3356 in trans. The results show that the dT+ phenotype requires a functional product encoded by STM 3356, and support the use of the PCR-based discrimination method for S. Paratyphi B and S. Java as the standard differentiation method. PMID- 17108270 TI - A novel real-time PCR to detect Chlamydia trachomatis in first-void urine or genital swabs. AB - Screening for Chlamydia trachomatis infections can be performed on urine samples and genital swabs using molecular techniques. A novel approach was developed that combined an automated extraction procedure, an automated liquid-handling system and real-time PCR to detect C. trachomatis from urine or swabs. This novel real time PCR approach was compared to the commercial Cobas Amplicor system on 628 specimens. In a retrospective analysis, 51 samples that tested positive using the Cobas assay were also positive with the real-time PCR, whereas the 49 samples negative with Cobas were also negative with the real-time PCR, for an overall agreement of 100 %. Among 528 prospective samples consecutively received at the authors' laboratory with a request for C. trachomatis PCR, five PCR reactions were inhibited when tested with Cobas. These five inhibited samples were found negative with the real-time PCR. Among the remaining 523 samples, 45 (8.6 %) were positive with both methods, 476 (91 %) were negative with both methods, and 2 (0.4 %) were positive with Cobas but negative with the real-time PCR. Thus, when considering Cobas as the gold standard, the overall agreement was 99.6 %, the sensitivity of the real-time PCR was 95.7 % and the specificity was 100 %. The two discrepant samples were retested in parallel and were found negative with both methods. When testing a batch of 25 samples, both reagent costs and laboratory technician time were reduced with the new technique (7.30 euros per sample and 134 min) compared to Cobas (11.20 euros per sample and 232 min). Moreover, due to reduced organizational constraints, the median time from sample reception to result was only 24 h using the automated platform. Overall, this novel real-time PCR approach exhibited an excellent specificity and a sensitivity similar to that of Cobas Amplicor PCR for the detection of C. trachomatis. Given its high throughput potential and low costs/laboratory technician time requirement, it may be useful for future use in large C. trachomatis screening programs. PMID- 17108271 TI - Identification of different clonal complexes and diverse amino acid substitutions in penicillin-binding protein 2 (PBP2) associated with borderline oxacillin resistance in Canadian Staphylococcus aureus isolates. AB - Borderline oxacillin-resistant Staphylococcus aureus (BORSA) exhibit oxacillin MIC values of 1-8 microg ml(-1), but lack mecA, which encodes the low-affinity penicillin-binding protein (PBP)2a. The relationship of the BORSA phenotype with specific genetic backgrounds was assessed, as well as amino acid sequence variation in the normal PBP2. Among 38 BORSA, 26 had a common PFGE profile of genomic DNA, and were multilocus sequence type (ST)25. The other isolates were genetically diverse. Complete pbp2 sequences were determined for three BORSA, corresponding to ST25, ST1 and ST47, which were selected on the basis of lacking blaZ-encoded beta-lactamase. The essential transpeptidase-domain-encoding segment of pbp2 was also sequenced from seven additional ST25 isolates. Amino acid substitutions occurred in the transpeptidase domain of all BORSA, irrespective of clonal type. A Gln(629)-->Pro substitution was common to all ST25 BORSA, but most could be distinguished from one another by additional unique substitutions in the transpeptidase domain. The ST1 and ST47 isolates also possessed unique substitutions in the transpeptidase domain. Plasmid-mediated expression of pbp2 from an ST25 or ST1 isolate in S. aureus RN6390 increased its oxacillin MIC from 0.25 to 4 microg ml(-1), while pbp2 from a susceptible strain, ATCC 25923, had no effect. Therefore, different amino acid substitutions in PBP2 of diverse BORSA lineages contribute to borderline resistance. The predominant ST25 lineage was not related to any of the five clonal complexes that contain meticillin-resistant S. aureus (MRSA), suggesting that ST25 cannot readily acquire mecA-mediated resistance. PMID- 17108272 TI - Genetic characterization of multidrug resistance in Shigella spp. from Japan. AB - This study characterized the genetic basis of antimicrobial resistance of a number of Shigella spp. isolated from humans from 2000 to 2004 in Hiroshima prefecture, Japan. A total of 26 isolates of Shigella spp. were included in this study. Antimicrobial susceptibility tests revealed high levels of resistance, especially to ampicillin, streptomycin, trimethoprim, tetracycline, nalidixic acid and ciprofloxacin. PCR and DNA sequencing were used for screening and characterization of antibiotic-resistance determinants. PCR sequencing analysis revealed the presence of only one type of class 1 integron in one isolate of Shigella sonnei. This class 1 integron was 1904 bp and contained two gene cassettes: a probable esterase/lipase (estX) and aadA1, which confers resistance to streptomycin and spectinomycin. Two types of class 2 integron were identified in this study. One was the classic type (2158 bp) and carried the three conserved resistance gene cassettes of the class 2 integron, dfrA1, sat1 and aadA1, which confer resistance to trimethoprim, streptothricin and streptomycin/spectinomycin, respectively. This type was detected in both Shigella sonnei (14 isolates) and Shigella flexneri (five isolates). The other type was shorter (1313 bp) and carried only two gene cassettes, dfrA1 and sat1. This integron was detected in a single isolate of Shigella sonnei. PFGE patterns showed limited diversity within clusters of the same species. Furthermore, an extended-spectrum beta-lactamase gene, bla(OXA-30), which confers resistance to ampicillin, was characterized in all isolates of Shigella flexneri except the oldest strain, which was isolated in 2000. Southern blot hybridization and conjugation experiments showed that bla(OXA 30) was located in the chromosome. PMID- 17108273 TI - Comparative analysis of Etest and broth microdilution method (AFST-EUCAST) for trends in antifungal drug susceptibility testing of Brazilian Cryptococcus neoformans isolates. AB - A prospective study was performed to evaluate the correlation between the proposed standard of the Antifungal Susceptibility Testing Subcommittee of the European Committee on Antibiotic Susceptibility Testing (AFST-EUCAST) (document 7.1) and the commercial system Etest for determining the MICs of flucytosine, amphotericin B, fluconazole, itraconazole and voriconazole for a collection of 100 clinical and environmental isolates of Cryptococcus neoformans. The agreements among Etest MICs within +/-2 log2 dilutions of AFST-EUCAST standard MICs were greater for flucytosine, fluconazole and voriconazole (76, 78 and 88 %, respectively) than for amphotericin B (5 %), the lowest agreement, and itraconazole (67 %). Overall, the correlation coefficients were statistically significant (P<0.05), and it is suggested that the Etest and AFST-EUCAST method are reliable alternatives and present good correlation for all drugs evaluated except amphotericin B. However, the observed differences related to MICs for susceptible, susceptible dose dependent and resistant strains between the methods suggest that it will be necessary to carry out further studies, including assessment of interlaboratory agreement and correlation of MICs by different methods with in vivo response. PMID- 17108274 TI - emm typing of invasive T28 group A streptococci, 1995-2004, Finland. AB - A total of 985 group A streptococcus (GAS) bacteraemia isolates collected in Finland during 1995-2004 were T-serotyped, and of these, 336 isolates of serotype T28 were subjected to further emm typing. The total number of isolates referred per year showed an increase within the study period, from 43 in 1995 to 130 in 2004. The annual incidence of invasive GAS (iGAS) bacteraemia showed a general increase during the study period, from 1.1 to 2.5 per 100 000 population. Serotype T28 remained among the most common serotypes, in addition to serotypes TB3264 and T1. The serotype T28 isolates were found to be distributed across six distinct emm types: emm28, emm77, emm53 (including subtypes 53.2 and 53.4), emm87, emm2 and emm4. The serotype distribution and the emm type distribution of serotype T28 fluctuated over time. Within the study period, the proportion of T28/emm28 isolates became the most prominent. During periods of low emm28 incidence, emm types 77 and 53 seemed to show a resurgence. emm typing revealed T28 isolates to be a genetically heterogeneous group harbouring a variety of distinct M proteins. This study confirms that T serotyping alone is not a sufficient method for epidemiological surveillance of iGAS. PMID- 17108275 TI - Use of sequence-based typing for investigation of a case of nosocomial legionellosis. AB - A fatal case of nosocomial legionellosis in a low prevalence region (Calgary, Alberta, Canada) prompted investigation into the source of infection. Hospital water systems contaminated with Legionella pneumophila have been shown to pose a risk to compromised patients. Typing of an L. pneumophila serogroup 1 strain isolated from the patient using sequence-based typing (SBT) and amplified fragment length polymorphism (AFLP) analysis linked it to a persistent and widespread strain isolated from the hospital water system establishing a nosocomial mode of acquisition. Different SBT and AFLP patterns were determined for non-epidemiologically linked cases and isolates from different hospitals. PMID- 17108276 TI - Rapid identification of clinically relevant Nocardia species using real-time PCR with SYBR Green and melting-curve analysis. AB - The objective of this study was to develop and evaluate a rapid new method of identifying clinically relevant Nocardia species. DNA extracted from different Nocardia strains was used in a real-time PCR assay with SYBR Green and melting curve analysis to identify Nocardia species. Ten control strains and four bacterial strains of closely related genera were employed, and samples from 28 patients were used. All Nocardia strains were identified correctly, and there was no cross-reaction with strains from genera closely related to Nocardia. The sensitivity and specificity of the method were 90 and 100 %, respectively. This method can be used to rapidly detect Nocardia species in culture, without cross reaction with other closely related genera. PMID- 17108277 TI - Listeria monocytogenes translocates throughout the digestive tract in asymptomatic sheep. AB - Ruminants are fed forage which is often contaminated with Listeria, and frequently shed Listeria monocytogenes with their faeces. The present study was designed to localize the sites of infection in the digestive tract concomitant with Listeria faecal excretion in a sheep model. Ten Listeria-free sheep were inoculated per os with a dose of 10(10) c.f.u. of a pathogenic L. monocytogenes strain. Listeria received by two of the ten animals were radiolabelled with (111)indium oxine. The dissemination of the Listeria was assessed by in vivo imaging, by culture of bacteria in the faeces, organs and digesta samples taken at slaughter on days 1, 2, 6, 10 and 14 post-inoculation, and by measuring gamma radioactivity of samples on day 6. It was shown that Listeria spread through the entire volume of the forestomachs within 4 h, and through the whole gastrointestinal tract (GIT) within 24 h. Faecal shedding of Listeria lasted 10 days. Rumen, duodenum, jejunum, ileum, caecum and colon walls and digesta, mesenteric lymph nodes, liver and spleen were temporarily infected. However, Listeria persisted for at least 14 days in rumen digesta and retropharyngeal lymph nodes, and at a relatively high level (1 x 10(4) c.f.u. g(-1)) in palatine tonsils. These findings suggest that L. monocytogenes can translocate from all parts of the GIT, with the rumen digesta, but not the gallbladder, serving as a reservoir. The results indicate that brief and low-level faecal excretion of L. monocytogenes is concomitant with a transitory asymptomatic infection in sheep. PMID- 17108278 TI - Yersinia enterocolitica isolates of differing biotypes from humans and animals are adherent, invasive and persist in macrophages, but differ in cytokine secretion profiles in vitro. AB - Previous epidemiological studies have demonstrated a potential link between the serotypes of Yersinia enterocolitica recovered from cattle, sheep and pigs and those isolated from human disease cases. Further studies utilizing amplified fragment length polymorphisms have shown a relationship at the genetic level between strains of biotypes 3 and 4 from humans and livestock, and also suggested that some biotype 1A isolates, classically defined as non-pathogenic, are closely related to biotype 3 and 4 isolates. This study sought to understand further the pathogenic potential of Y. enterocolitica isolates from livestock in Great Britain. A range of surrogate in vitro models, such as invasion of epithelial tissue cultures, survival in cultured macrophages and cytokine secretion response, was employed to assess the pathogenicity of 88 strains. The results suggested that all isolates examined were capable of adhering to and invading epithelial cells and of surviving within macrophages. However, the inflammatory response of the infected macrophages differed with the infecting Y. enterocolitica subtype, with the response to pathogenic biotype 3 and 4 isolates different to that observed with biotype 1A isolates, and with the biotype 3 O : 5,27 isolates recovered exclusively from animals. Infections of porcine tissue also suggested the possibility of host-tissue tropism within Y. enterocolitica subtypes. PMID- 17108279 TI - Humoral and cellular immune responses in sheep immunized with a 22 kilodalton exported protein of Mycobacterium avium subspecies paratuberculosis. AB - An immunogenic 22 kilodalton exported Mycobacterium avium subspecies paratuberculosis (MAP) lipoprotein (P22) was previously identified, and found to belong to the LppX/LprAFG family of mycobacterial lipoproteins. N-terminal polyhistidine-tagged P22 was produced and purified from Escherichia coli. Antibody recognition of P22, and interferon-gamma (IFN-gamma) responses in vitro using blood from a sheep vaccinated with Neoparasec, confirmed its immunogenicity. To evaluate the immunogenicity of P22 in vivo, five sheep were immunized with a single dose containing 0.8 mg recombinant P22 protein in adjuvant. Blood was collected at 4, 13 and 29 weeks post-immunization (p.i.) and tested for anti-P22 antibodies and P22-specific IFN-gamma production. P22 specific antibodies were detected by Western blot analysis in all five Neoparasec immunized sheep at the three time points. Three out of five P22-immunized sheep produced P22-specific antibodies for up to 13 weeks p.i., and two gave a response at 29 weeks p.i. Recombinant P22 was able to stimulate significant IFN-gamma production in blood of P22-immunized sheep at 13 and 29 weeks p.i. Recombinant P22 also elicited an IFN-gamma response in blood of sheep immunized with Neoparasec. PMID- 17108280 TI - Recovery of interfering and beta-lactamase-producing bacteria from group A beta haemolytic streptococci carriers and non-carriers. AB - The purpose of this study was to compare the frequency of recovery of aerobic and anaerobic organisms with interfering capability against group A beta-haemolytic streptococci (GABHS) and beta-lactamase-producing bacteria (BLPB) from the tonsils of GABHS carriers and non-carriers. The presence of aerobic and anaerobic bacteria capable of such interference in vitro was evaluated in cultures obtained from the tonsils of 20 healthy children who were non-GABHS carriers and 20 who were GABHS carriers, and also from 20 children who were asymptomatic after completing a course of penicillin for acute GABHS pharyngo-tonsillitis (PT) and were non-GABHS carriers and 20 who were GABHS carriers. In healthy children, 32 interfering isolates were recovered from 16 non-GABHS carriers (1.6 per child) and 13 were isolated from 7 GABHS carriers (0.65 per child) (P<0.001). In children who had suffered acute GABHS PT, 26 interfering organisms were recovered from 15 non-GABHS carriers (1.3 per child) and 8 were isolated from 5 GABHS carriers (0.4 per child) (P<0.005). In healthy children, 13 BLPB were recovered from 5 non-GABHS carriers and 13 were isolated from 6 GABHS carriers. In children who had suffered acute GABHS PT, 14 BLPB were recovered from 5 (25 %) non-GABHS carriers and 32 were isolated from 17 (85 %) GABHS carriers (P<0.05). It was demonstrated in this study that there was a higher rate of recovery of aerobic and anaerobic organisms capable of interfering with GABHS in non-GABHS carriers than in GABHS carriers. This was observed in all GABHS non-carriers and included healthy children as well as those recently treated for symptomatic GABHS PT with penicillin that failed to eradicate GABHS. A higher rate of recovery of BLPB was observed only in GABHS carriers who were treated with penicillin for GABHS PT. PMID- 17108281 TI - Mycobacterium kansasii olecranon bursitis. AB - A case is reported of a post-traumatic olecranon bursitis caused by Mycobacterium kansasii following an injury sustained in a public swimming pool. It responded to surgical debridement and combined rifampicin, isoniazid, pyrazinamide and ethambutol antimicrobial therapy. A literature search was performed and a treatment regimen for this uncommon condition is suggested. PMID- 17108282 TI - Influence of the fsr locus on biofilm formation by Enterococcus faecalis lacking gelE. PMID- 17108283 TI - Conditional production of acyl-homoserine lactone-type quorum-sensing signals in clinical isolates of enterobacteria. PMID- 17108284 TI - Interaction of Vibrio cholerae O139 with an intestinal parasite, Entamoeba histolytica. PMID- 17108285 TI - Enterotoxigenic Clostridium perfringens and sporadic diarrhoea: a study from an Indian tertiary care hospital. PMID- 17108286 TI - Detecting immunoglobulin M antibodies against microsporidian Encephalitozoon cuniculi polar tubes in sera from healthy and human immunodeficiency virus infected persons in Japan. AB - Encephalitozoon cuniculi, a spore-forming obligate intracellular parasitic pathogen belonging to the phylum Microsporidia, has a unique and highly specialized organelle called the polar tube. Using an enzyme immunostaining assay in which germinated E. cuniculi spores were coated onto plastic surfaces, we tested healthy and human immunodeficiency virus (HIV)-infected individuals in Japan for anti-polar tube antibodies of each immunoglobulin (Ig) class. Anti polar tube IgG was detected in just 4 of 380 healthy individuals; no anti-polar tube IgA was detected in any individuals; however, unexpectedly, anti-polar tube IgM antibodies were detected in 138 individuals (36%). When the healthy individuals were grouped by age, the highest rate of positivity to anti-polar tube IgM antibodies was seen in individuals aged 20 years old or younger. Fifty nine percent (24/41) of the individuals aged 20 years or younger were anti-polar tube IgM antibody positive. This rate tended to decrease among individuals in older age groups. However, no anti-polar tube IgM antibodies were detected in 21 HIV-infected persons who were younger than 30 years of age and who had CD4 cell levels below 250/mul. These seroepidemiological results clearly indicate that circulating anti-polar tube IgM antibodies that are capable of strongly reacting with filaments extruded from geminated spores exist and suggest that such antibodies may play a part in protective immunity. PMID- 17108287 TI - Naturally occurring anti-Escherichia coli protein antibodies in the sera of healthy humans cause analytical interference in a recombinant nucleocapsid protein-based enzyme-linked immunosorbent assay for serodiagnosis of severe acute respiratory syndrome. AB - We reported the analytical interference of anti-Escherichia coli protein (EP) antibodies in human sera and residual EP in a recombinant nucleocapsid protein based enzyme-linked immunosorbent assay as a possible source of false positives in severe acute respiratory syndrome serodiagnosis. The rate of false positives was significantly reduced by adding mouse anti-EP antiserum in the blocking step. PMID- 17108288 TI - Borrelia burgdorferi spirochetes that harbor only a portion of the lp28-1 plasmid elicit antibody responses detectable with the C6 test for Lyme disease. AB - Detection of antibody to C6, a peptide that reproduces the sequence of the sixth invariable region within the central domain of the VlsE protein of Borrelia burgdorferi, is used currently for the serologic diagnosis of Lyme disease in humans. B. burgdorferi isolates taken from infected humans can be categorized into specific genetic subtypes (designated RST1, -2, and -3) by restriction fragment length polymorphisms in the 16S to 23S rRNA spacer sequence. Many of these, usually categorized as RST2, retain only segments of the linear plasmid lp28-1, which encodes VlsE. The VlsE genetic region is retained, but altered expression of this molecule could affect diagnosis by the C6 enzyme-linked immunosorbent assay (ELISA). Serum samples from patients infected with each of the three genotypes and from mice infected with three RST2 isolates were tested with the C6 ELISA. Such isolates elicited marked C6 responses in infected mice. The sensitivity of C6 antibody detection in patients infected with RST2 spirochetes was statistically indistinguishable from detection of RST1 and RST3 infections. These findings demonstrate that diagnosis by C6 ELISA remains effective for infection with all B. burgdorferi genotypes, including those with incomplete lp28-1 plasmids. PMID- 17108289 TI - Optimized FaeG expression and a thermolabile enterotoxin DNA adjuvant enhance priming of an intestinal immune response by an FaeG DNA vaccine in pigs. AB - One of the problems hindering the development of DNA vaccines is the relatively low immunogenicity often seen in humans and large animals compared to that in mice. In the present study, we tried to enhance the immunogenicity of a pcDNA1/faeG19 DNA vaccine in pigs by optimizing the FaeG expression plasmid and by coadministration of the plasmid vectors encoding the A and B subunits of the Escherichia coli thermolabile enterotoxin (LT). The insertion of a Kozak sequence and optimization of vector (cellular localization and expression) and both vector and codon usage were all shown to enhance in vitro FaeG expression compared to that of pcDNA1/faeG19. Subsequently, pcDNA1/faeG19 and the vector-optimized and the vector-codon-optimized construct were tested for their immunogenicity in pigs. In line with the in vitro results, antibody responses were better induced with increasing expression. The LT vectors additionally enhanced the antibody response, although not significantly, and were necessary to induce an F4-specific cellular response. These vectors were also added because LT has been described to direct the systemic response towards a mucosal immunoglobulin A (IgA) response in mice. Here, however, the intradermal FaeG DNA prime-oral F4 boost immunization resulted in a mainly systemic IgG response, with only a marginal but significant reduction in F4+ E. coli fecal excretion when the piglets were primed with pWRGFaeGopt and pWRGFaeGopt with the LT vectors. PMID- 17108290 TI - Bacterial interactions and the microevolution of cytochrome bd: implications for pathogenesis. PMID- 17108291 TI - Microevolution of cytochrome bd oxidase in Staphylococci and its implication in resistance to respiratory toxins released by Pseudomonas. AB - Pseudomonas aeruginosa and Staphylococcus aureus are opportunistic pathogens and frequently coinfect the lungs of cystic fibrosis patients. P. aeruginosa secretes an arsenal of small respiratory inhibitors, like pyocyanin, hydrogen cyanide, or quinoline N-oxides, that may act against the commensal flora as well as host cells. Here, we show that with respect to their susceptibility to these respiratory inhibitors, staphylococcal species can be divided into two groups: the sensitive group, comprised of pathogenic species such as S. aureus and S. epidermidis, and the resistant group, represented by nonpathogenic species such as S. carnosus, S. piscifermentans, and S. gallinarum. The resistance in the latter group of species was due to cydAB genes that encode a pyocyanin- and cyanide-insensitive cytochrome bd quinol oxidase. By exchanging cydB in S. aureus with the S. carnosus-specific cydB, we could demonstrate that CydB determines resistance. The resistant or sensitive phenotype was based on structural alterations in CydB, which is part of CydAB, the cytochrome bd quinol oxidase. CydB represents a prime example of both microevolution and the asymmetric pattern of evolutionary change. PMID- 17108292 TI - Could low-level background exposure to persistent organic pollutants contribute to the social burden of type 2 diabetes? PMID- 17108293 TI - The United Nations and road safety: what is the connection? PMID- 17108294 TI - A glossary for the social epidemiology of work organisation: part 2 Terms from the sociology of work and organisations. PMID- 17108295 TI - The irony of a rich country: issues of financial access to and availability of healthy food in the Republic of Ireland. AB - OBJECTIVE: To determine the weekly financial cost of a diet as recommended by national policy in two parents with two children, single parents with one child and single old people with low income, and begin to identify, in a rich country context, variation in food item availability, price and household purchasing capacity. DESIGN: Food baskets were developed based on national dietary recommendations and purchasing patterns of these household groups. National-level prices of each food were identified, as well as pricing across a representative selection of Irish retail outlet types. Basket costs were assessed relative to the financial capacity of household type. RESULTS: The types of retail outlets in which low-income groups tend to shop do not carry many own brand items and is less likely to stock healthy options, but when they do, they are more expensive than in other outlets. Single parents with one child, two adults with two children and single older people would have to spend 80%, 69% and 38%, respectively, of their weekly household income to purchase the food basket based on economy-line products. CONCLUSIONS: Financial access to and availability of healthy food options must be considered through a national policy cognisant of basic human needs for healthy living. This research provides evidence on the direct costs of healthy eating for policy and planning to ensure not only financial capacity but also to guarantee that affordable healthy food choices are physically available to all groups in society. PMID- 17108296 TI - Women's health in a rural community in Kerala, India: do caste and socioeconomic position matter? AB - OBJECTIVES: To examine the social patterning of women's self-reported health status in India and the validity of the two hypotheses: (1) low caste and lower socioeconomic position is associated with worse reported health status, and (2) associations between socioeconomic position and reported health status vary across castes. DESIGN: Cross-sectional household survey, age-adjusted percentages and odds ratios, and multilevel multinomial logistic regression models were used for analysis. SETTING: A panchayat (territorial decentralised unit) in Kerala, India, in 2003. PARTICIPANTS: 4196 non-elderly women. OUTCOME MEASURES: Self perceived health status and reported limitations in activities in daily living. RESULTS: Women from lower castes (scheduled castes/scheduled tribes (SC/ST) and other backward castes (OBC) reported a higher prevalence of poor health than women from forward castes. Socioeconomic inequalities were observed in health regardless of the indicators, education, women's employment status or household landholdings. The multilevel multinomial models indicate that the associations between socioeconomic indicators and health vary across caste. Among SC/ST and OBC women, the influence of socioeconomic variables led to a "magnifying" effect, whereas among forward caste women, a "buffering" effect was found. Among lower caste women, the associations between socioeconomic factors and self-assessed health are graded; the associations are strongest when comparing the lowest and highest ratings of health. CONCLUSIONS: Even in a relatively egalitarian state in India, there are caste and socioeconomic inequalities in women's health. Implementing interventions that concomitantly deal with caste and socioeconomic disparities will likely produce more equitable results than targeting either type of inequality in isolation. PMID- 17108297 TI - Inequity in access to dental care services explains current socioeconomic disparities in oral health: the Swedish National Surveys of Public Health 2004 2005. AB - OBJECTIVE: To analyse the effects of socioeconomic disadvantage on access to dental care services and on oral health. Design, setting and outcomes: Cross sectional data from the Swedish National Surveys of Public Health 2004 and 2005. Outcomes were poor oral health (self-rated oral health and symptoms of periodontal disease) and lack of access to dental care services. A socioeconomic disadvantage index (SDI) was developed, consisting of social welfare beneficiary, being unemployed, financial crisis and lack of cash reserves. PARTICIPANTS: Swedish population-based sample of 17 362 men and 20 037 women. RESULTS: Every instance of increasing levels of socioeconomic disadvantage was associated with worsened oral health but, simultaneously, with decreased utilisation of dental care services. After adjusting for age, men with a mild SDI compared with those with no SDI had 2.7 (95% confidence interval (CI) 2.5 to 3.0) times the odds for self-rated poor oral health, whereas odds related to severe SDI were 6.8 (95% CI 6.2 to 7.5). The corresponding values among women were 2.3 (95% CI 2.1 to 2.5) and 6.8 (95% CI 6.3 to 7.5). Nevertheless, people with severe socioeconomic disparities were 7-9 times as likely to refrain from seeking the required dental treatment. These associations persisted even after controlling for living alone, education, occupational status and lifestyle factors. Lifestyle factors explained only 29% of the socioeconomic differences in poor oral health among men and women, whereas lack of access to dental care services explained about 60%. The results of the multilevel regression analysis indicated no additional effect of the administrative boundaries of counties or of municipalities in Sweden. CONCLUSIONS: Results call for urgent public health interventions to increase equitable access to dental care services. PMID- 17108298 TI - Cross-cultural adaptation of a tobacco questionnaire for Punjabi, Cantonese, Urdu and Sylheti speakers: qualitative research for better clinical practice, cessation services and research. AB - OBJECTIVE: To develop cross-culturally valid and comparable questionnaires for use in clinical practice, tobacco cessation services and multiethnic surveys on tobacco use. METHODS: Key questions in Urdu, Cantonese, Punjabi and Sylheti on tobacco use were compiled from the best existing surveys. Additional items were translated by bilingual coworkers. In one-to-one and group consultations, lay members of the Pakistani, Chinese, Indian Sikh and Bangladeshi communities assessed the appropriateness of questions. Questionnaires were developed and field tested. Cross-cultural comparability was judged in a discussion between the researchers and coworkers, and questionnaires were finalised. Questionnaires in Cantonese (written and verbal forms differ) and Sylheti (no script in contemporary use) were written as spoken to avoid spot translations by interviewers. RESULTS: The Chinese did not use bidis, hookahs or smokeless tobacco, so these topics were excluded for them. It was unacceptable for Punjabi Sikhs to use tobacco. For the Urdu speakers and Sylheti speakers there was no outright taboo, particularly for men, but it was not encouraged. Use of paan was common among women and men. Many changes to existing questions were necessary to enhance cultural and linguistic appropriateness-for example, using less formal language, or rephrasing to clarify meaning. Questions were modified to ensure comparability across languages, including English. CONCLUSION: Using theoretically recommended approaches, a tobacco-related questionnaire with face and content validity was constructed for Urdu, Punjabi, Cantonese and Sylheti speakers, paving the way for practitioners to collect more valid data to underpin services, for sounder research and ultimately better tobacco control. The methods and lessons are applicable internationally. PMID- 17108299 TI - The geography of despair among 15-44-year-old men in England and Wales: putting suicide on the map. AB - OBJECTIVE: To investigate the spatial patterning and possible contributors to the geographical distribution of suicide among 15-44- year-old men. DESIGN: Small area analysis and mapping of geo-coded 1988-94 suicide mortality data and 1991 census data using random-effects smoothing. SETTING: 9265 electoral wards in England and Wales (mean population of men aged 15-44- years: about 1220). MAIN RESULTS: Two main patterns emerged: (a) in all of the 10 most densely populated cities studied, suicide showed a "bull's-eye" pattern with rates highest in the inner-city areas and, in some cases, low rates in the peripheries, and (b) suicide rates were high in coastal areas, particularly those in more remote regions. Possible indicators of social fragmentation, such as the proportion of single-person households in an area, were most strongly and consistently associated with rates of suicide in both urban and rural areas. Levels of unemployment and long-term illness accounted for some of the coastal patterning. Although characteristics of areas accounted for more than half of the observed variability, substantial between-area variability in rates remained unexplained. CONCLUSIONS: The area characteristics investigated here did not fully account for the higher suicide rates observed in the most rural or remote areas. Alongside social and economic aspects, rural life itself may have an independent effect on the risk of suicide. A greater understanding of local geographies of suicide, and particularly the possible interactions between characteristics of people and their environments, might assist the design of prevention strategies that target those areas (and their characteristics) where risk is concentrated. PMID- 17108300 TI - Alcohol misuse in older people: heavy consumption and protean presentations. AB - BACKGROUND: Alcohol misuse, especially binge drinking in young people, and alcoholic liver disease are major public health concerns. However, alcohol misuse in older people is underestimated and often goes undetected. OBJECTIVE: To document alcohol consumption and clinical presentation of alcohol misuse in hospital inpatients aged >or=60 years. METHODS: 208 inpatients aged >or=60 years, referred to the alcohol liaison nurse between 1998 and 2003 at the Royal Bolton Hospital, Bolton, UK, were assessed for sex, alcohol intake, primary and secondary reasons for admission, and other concurrent health problems and death. RESULTS: 90% of men drank >21 units weekly and 93% of women drank >14 units weekly. Median weekly alcohol intake was 78.5 units for men and 47 units for women. Acute intoxication, falls, circulatory problems and alcoholic liver disease were the main primary reasons for admission. Neglect or malnutrition, alcoholic liver disease and hypertension were the main secondary reasons and concurrent health problems. 30% of patients died between 1998 and 2003. CONCLUSION: In inpatients aged >or=60 years who were referred to the alcohol liaison nurse in a district general hospital, heavy alcohol consumption, often to very high levels, was characteristic in both men and women and was associated with a wide variety of primary and secondary clinical presentations, including death. PMID- 17108301 TI - Area deprivation, individual factors and low birth weight in England: is there evidence of an "area effect"? AB - OBJECTIVE: To explore the relationship between low and very low birth weights, mother's age, individual socioeconomic status and area deprivation. DESIGN: Analysis of the incidence of low and very low birth weights by area deprivation, maternal age, social class of household and estimated income. SETTING: England 1996-2000. SUBJECTS: 2 894 440 singleton live births and the 10% sample of these births for which parents' individual-level socioeconomic measures were coded. RESULTS: Social class, estimated household income, lone-parenthood and mother's age were all associated with the risk of low and very low birth weight. Even when controlling for these individual level factors, area income deprivation was significantly associated with low and very low birth weight (p<0.00). For low birth weight there was a significant interaction between area income deprivation and mother's age. For very young mothers, the area effect was non-significant (p<0.37). For older mothers, particularly those aged 30-34 years, it was stronger (p<0.00). As a result, mothers aged <18 years, although at relatively high risk of low birth weight irrespective of area income deprivation, were actually at slightly lower risk than mothers aged >40 years in the most deprived areas. CONCLUSIONS: For all but very young mothers, there seems to be a negative effect on birth weight from living in areas of income deprivation, whatever their individual circumstances. PMID- 17108302 TI - Neighbourhood mortality inequalities in New York City, 1989-1991 and 1999-2001. AB - OBJECTIVES: To examine whether inequalities in mortality across socioeconomically diverse neighbourhoods changed alongside the decline in mortality observed in New York City between 1990 and 2000. DESIGN: Cross-sectional analysis of neighbourhood-level vital statistics. SETTING: New York City, 1989-1991 and 1999 2001. MAIN RESULTS: In both poor and wealthy neighbourhoods, age-adjusted mortality for most causes declined between the time periods, although mortality from diabetes increased. Relative inequalities decreased slightly-largely in the under 65 years population-although all-cause rates in 1999-2001 were still 50% higher, and rates of years of potential life lost before age 65 years were 150% higher, in the poorest communities than in the wealthiest ones (relative index of inequality 1.7 and 3.3, respectively). The relative index of inequality for mortality from AIDS increased from 4.7 to 13.9. Over 50% of the excess mortality in the poorest neighbourhoods in 1999-2001 was due to cardiovascular disease, AIDS and cancer. CONCLUSIONS: In New York City, despite substantial declines in absolute mortality and rate differences between poor and wealthy neighbourhoods, great relative socioeconomic inequalities in mortality persist. PMID- 17108303 TI - Provincial income inequality and self-reported health status in China during 1991 7. AB - BACKGROUND: The relationship between income inequality and health has been widely explored. Today there is some evidence suggesting that good health is inversely related to income inequality. After the economic reforms initiated in the early 1980s, China experienced one of the fastest-growing income inequalities in the world. The state of China in the 1990s is focussed on and possible effects of provincial income inequality on individual health status are explored. METHODS: A multilevel regression model is used to analyse the data collected in 1991, 1993 and 1997 from nine provinces included in the China Health and Nutrition Survey. The effects of provincial Gini coefficients on self-rated health in each year are evaluated by two logistic regressions estimating the odds ratios of reporting poor or fair health. The patterns of this effect are compared among the survey years and also among different demographic groups. RESULTS: The analyses show an independent effect of income inequality on self-reported health after adjusting for individual and household variables. Furthermore, the effect of income distribution is not attenuated when household income and provincial gross domestic product per capita are included in the model. The results show that there is an increased risk of about 10-15% on average for fair or poor health for people living in provinces with greater income inequalities compared with provinces with modest income inequalities. CONCLUSIONS: In China, societal income inequality appears to be an important determinant of population health during 1991-7. PMID- 17108304 TI - Gender differences in work-home interplay and symptom perception among Swedish white-collar employees. AB - OBJECTIVE: To analyse gender differences in paid and unpaid workload and symptoms in matched groups of Swedish white-collar workers with children. DESIGN AND SETTING: Paid and unpaid workload and perceived stress from paid work, conflict between demands and control over household work were measured by a total workload (TWL) questionnaire. Some symptoms were rated with regard to frequency and severity as a measure of health. Cross-sectional analyses were performed. PARTICIPANTS: Matched groups of male (n = 440) and female (n = 529) well-educated white-collar workers in full-time employment, aged 32-58 years and living with children in the home. RESULTS: Women in higher positions in Sweden are healthier than the average population of women, but report more symptoms than men in the same position as well as more stress from paid work, more conflict between demands and a greater TWL. However, women also reported more control over household duties and TWL was not associated with more symptoms. The men were mainly focused on their paid work role and perhaps even more so than men in the general population as they were fairly resistant to feelings of conflicting demands. CONCLUSION: Even among matched groups of full-time employed, well educated men and women, traditional gender differences in division of responsibilities and time allocation were found. Even though the women were healthy at this stage, they might risk future ill health, owing to high workload, stress and feelings of conflicting demands. PMID- 17108305 TI - The contribution of smoking to socioeconomic differentials in mortality: results from the Melbourne Collaborative Cohort Study, Australia. AB - OBJECTIVE: To assess the contribution of smoking to the inverse association of mortality with years of formal education in men in Australia. DESIGN: Data were obtained from a prospective cohort study that included 17 049 men in Melbourne recruited from 1990 to 1994, most of whom were aged between 40 and 69 years at baseline. The outcome measured was all-cause mortality. The contribution of smoking to socioeconomic status differentials was estimated by including smoking as a variable in a Cox's proportional hazards model that also included education and other potential confounding variables. RESULTS: In men, the association between education and mortality was attenuated after adjustment for smoking, and the aetiological fraction for low levels of education was reduced from 16.5% to 10.6%. CONCLUSIONS: In men, smoking contributes substantially to socioeconomic differentials in mortality. Effective policies and interventions that target smoking among socially disadvantaged groups may substantially reduce socioeconomic differentials in health. PMID- 17108306 TI - Accumulation of adverse socioeconomic position over the entire life course and the risk of myocardial infarction among men and women: results from the Stockholm Heart Epidemiology Program (SHEEP). AB - BACKGROUND: Accumulation of adverse socioeconomic position over the life course is assumed to increase the risk of myocardial infarction. OBJECTIVE: To analyse in detail whether the accumulation of adverse socioeconomic position over the life course increases the risk of myocardial infarction, using yearly information on individual socioeconomic position from birth to disease onset. DESIGN: Case control study of risk factors for incident myocardial infarction (Stockholm Heart Epidemiology Program). SETTING: All Swedish citizens born during 1922-49 and living in Stockholm County during 1992-4. PARTICIPANTS: 550 female and 1204 male patients and 777 female and 1538 male controls. Every year in manual work was added to calculate a proportion of the whole life course spent in adverse socioeconomic position. RESULTS: With increasing proportion of life spent in adverse socioeconomic position, we found an increasing risk of myocardial infarction. The relative risk of myocardial infarction was 2.36 (95% confidence interval (CI) = 1.79 to 3.11) for men and 2.54 (95% CI = 1.70 to 3.78) for women who, over the entire life course, had always been in adverse socioeconomic position compared with those who had never been in adversity. We also found a strong increase in risk from being in adversity for only a few years, indicating important selection processes. CONCLUSIONS: Accumulated experience of adverse socioeconomic position over the entire life course increases the risk of myocardial infarction for men and women, but it is not a pure accumulation process as "how" and "when" the accumulation occurs also seems to have a role. The accumulation effect is partly mediated by the acquisition of health-damaging experiences. PMID- 17108307 TI - Influence of socioeconomic status on clinical outcomes and quality of life after percutaneous coronary intervention. AB - OBJECTIVES: To determine whether socioeconomic status (SES) influences clinical outcomes and quality of life after percutaneous coronary intervention (PCI). DESIGN: Prospective observational study. SETTING: Two interventional cardiac centres. PARTICIPANTS: 1346 consecutive patients undergoing PCI over a 12-month period. OUTCOMES: Self reported health-related quality of life (HRQoL; EuroQol-5 Dimensions (EQ-5D); EuroQol Visual Analogue Scale (EQ-VAS)), repeat angiography, revascularisation, hospital admission, myocardial infarction and death within 12 months, by SES derived using postal address code. MAIN RESULTS: No significant differences were found between patients with high and low SES in the occurrence of repeat angiography (p = 0.55), repeat revascularisation (PCI, p = 0.81, CAEG, p = 0.27), total cardiac hospitalisation (p = 0.10), myocardial infarction (p = 0.97) or death 12 months after PCI (p = 0.88). Non-procedure-related readmissions were higher in patients with low SES (18.6% v 13.7%; p = 0.025). After adjustment for confounding factors, patients with low SES had lower HRQoL scores at baseline (95% CI for difference 0.01 to 0.14; p = 0.003) and at 12 months (95% CI 0.07 to 0.17; p<0.001) compared with those with high SES. CONCLUSIONS: Clinical outcomes were similar for patients in different SES groups. Patients with low SES had considerably more non-procedure-related readmissions and lower quality-of-life scores. Future studies on HRQoL after coronary revascularisation should take account of these important differences related to SES. PMID- 17108308 TI - Measuring inequalities in health: the case for healthy life expectancy. AB - OBJECTIVE: To evaluate healthy life expectancy (HLE) as a measure of health inequalities by comparing geographical and area-based deprivation-related inequalities in healthy and total life expectancy (TLE). DESIGN: Life table analysis based on ecological cross-sectional data. SETTING AND POPULATION: Council area quarters and postcode sector-based deprivation fifths in Scotland. MAIN OUTCOME MEASURES: Expectation of life in good self-assessed general health, or free from limiting long-term illness, and TLE, for females and males at birth. RESULTS: Women in Scotland have a life expectation of 70.3 years in good health, 61.6 years free from limiting long-term illness, and a TLE of 78.9 years. Comparable figures for men are 66.3, 58.6 and 73.5 years. TLE and HLE decrease with increasing area deprivation. Differences are substantially wider for HLE. A 4.7-year difference is seen in TLE between women living in the most and least deprived fifth of areas. The difference in HLE is 10.7 years in good health and 11.6 years free from limiting long-term illness. The degree of deprivation related inequality in HLE is 2.5 times wider for women and 1.8 times wider for men than in TLE. CONCLUSIONS: Differences in TLE underestimate health inequalities substantially. By including morbidity and mortality, HLE reflects the excess burden of ill health experienced by disadvantaged populations better. Inequalities in length of life and health status during life should be taken into account while monitoring inequalities in population health. PMID- 17108309 TI - Linkage of survey data with district-level lung cancer registrations: a method of bias reduction in ecological studies. AB - OBJECTIVE: To investigate a stratified ecological method for reducing ecological bias in studies that use aggregate data, by incorporating information on individual-level risk factors into the analysis. DESIGN: Cross-sectional study investigating associations between socioeconomic risk factors and lung cancer in the north of England, using 1991 UK Census Small Area Statistics and Sample of Anonymised Records with lung cancer registrations from three regional cancer registries for 1993-6. SETTING AND PATIENTS: 92 local authority districts in the north of England containing over three million people aged 45-74 years. RESULTS: Generally, groups considered more socioeconomically disadvantaged had an increased risk of lung cancer across districts. In the standard ecological analysis, effects for non-car ownership, social class III manual, social class IV/V and socioeconomic inactivity were insignificant, suggesting ecological bias. In the stratified ecological analysis these effects became significant (rate ratio (RR) 2.23, 95% confidence interval (CI) 1.79 to 2.78, p<0.001; RR 1.35, 95% CI 1.04 to 1.74, p = 0.022; RR 2.36, 95% CI 1.86 to 2.99, p<0.001; and RR 0.72, 95% CI 0.53 to 0.98, p = 0.039, respectively), and spuriously large positive effects for the social class III non-manual (RR 20.29) and unemployment groups (RR 147.53) reduced to a more reasonable level (RR 1.92, 95% CI 1.46 to 2.52, p<0.001; and RR 2.36, 95% CI 1.22 to 4.55, p = 0.011, respectively). CONCLUSIONS: Stratified ecological analysis incorporating information on individual-level covariates reduced the bias seen in a standard ecological analysis. The method is straightforward to apply and allows the linkage of health data with data from any large-scale complex survey where district of residence is known. PMID- 17108310 TI - Correlation between incidences of hospitalised accidental burns and deprivation scores: an ecological study. PMID- 17108311 TI - Variation in food quality may threaten the validity of epidemiological studies in China. PMID- 17108312 TI - Lupus nephritis: toll the trigger! PMID- 17108313 TI - Prevention of microalbuminuria in type 2 diabetes: millimeters or milligrams? PMID- 17108314 TI - Bone marrow-derived cells are sufficient and necessary targets to mediate glomerulonephritis and vasculitis induced by anti-myeloperoxidase antibodies. AB - Clinical and experimental evidence indicate that ANCA cause pauci-immune necrotizing and crescentic glomerulonephritis (NCGN) and systemic small vessel vasculitis in humans. One of the major target antigens for ANCA is myeloperoxidase (MPO). An animal model that closely resembles the human disease is induced by intravenous injection of anti-MPO IgG into mice. The likely primary pathogenic targets for the anti-MPO IgG are circulating neutrophils and monocytes, although other cells have been implicated, including endothelial cells and epithelial cells. Herein is reported a new model for anti-MPO-mediated glomerulonephritis and vasculitis that further documents the pathogenic potential of ANCA and demonstrates that bone marrow (BM)-derived cells are sufficient targets to cause anti-MPO disease in the absence of MPO in other cell type. MPO knockout (Mpo-/-) mice that were immunized with mouse MPO were exposed to irradiation and received a transplant of Mpo+/+ or Mpo-/- BM. Engraftment in mice with circulating anti-MPO resulted in development of pauci-immune NCGN in all mice and pulmonary capillaritis and splenic necrotizing arteritis in some. Anti MPO IgG also was introduced intravenously into chimeric mice by transplantation of Mpo+/+ BM into irradiated Mpo-/- mice or Mpo-/- BM into irradiated Mpo+/+ mice. Chimeric Mpo-/- mice with circulating MPO-positive neutrophils developed NCGN, whereas chimeric Mpo+/+ mice with circulating MPO-negative neutrophils did not, thereby indicating that BM-derived cells are not only sufficient but also necessary for induction of anti-MPO disease. This novel animal model further documents ANCA IgG interactions with neutrophils as a cause of ANCA-associated glomerulonephritis and vasculitis. PMID- 17108315 TI - Pathogenesis and prevention of bone loss in patients who have kidney disease and receive long-term immunosuppression. AB - The coexistence of kidney disease with a need for immunosuppressive therapy leads to the convergence of several threats to bone. These comprise general effects of the primary disease, e.g., inflammatory state, more specific effects of acute renal failure or chronic kidney disease, and effects of therapies. Multisystem inflammatory disease that requires immunosuppression is associated frequently with kidney damage, and any reduction of kidney function that takes the patient into or beyond chronic kidney disease stage 2 for more than a short time is likely to have a negative impact on bone health. Bone mineral density frequently is low and fracture rates are high, although correlations often are poor. Chronic inflammation leads to local and systemic imbalance between bone formation and resorption. Upregulation of NF-kappabeta ligand (RANKL) and variable downregulation of osteoprotegerin are implicated, and bone health may improve in response to treatment of the inflammatory state. Certain immunosuppressive agents, especially glucocorticoids and calcineurin inhibitors, contribute further to bone loss. Antiresorptive agents such as bisphosphonates are used widely and, although able to prevent loss of bone mineral density, have uncertain effects on fracture rates. Augmentation of anabolic activity is desirable but elusive. Synthetic parathyroid hormone is untested but has potential. Manipulation of the RANKL/osteoprotegerin system now is feasible using antibodies to RANKL or synthetic osteoprotegerin. In the future, manipulation of the calcium-sensing receptor using calcimimetic or calcilytic agents may allow the anabolic effects of parathyroid hormone to be harnessed to good effect. With all of these therapies, it will be important to assess response in relation to important clinical end points such as fracture. PMID- 17108316 TI - Early embryonic renal tubules of wild-type and polycystic kidney disease kidneys respond to cAMP stimulation with cystic fibrosis transmembrane conductance regulator/Na(+),K(+),2Cl(-) Co-transporter-dependent cystic dilation. AB - Metanephric organ culture has been used to determine whether embryonic kidney tubules can be stimulated by cAMP to form cysts. Under basal culture conditions, wild-type kidneys from embryonic day 13.5 to 15.5 mice grow in size and continue ureteric bud branching and tubule formation over a 4- to 5-d period. Treatment of these kidneys with 8-Br-cAMP or the cAMP agonist forskolin induced the formation of dilated tubules within 1 h, which enlarged over several days and resulted in dramatically expanded cyst-like structures of proximal tubule and collecting duct origin. Tubule dilation was reversible upon withdrawal of 8-Br-cAMP and was inhibited by the cAMP-dependent protein kinase inhibitor H89 and the cystic fibrosis transmembrane conductance regulator (CFTR) inhibitor CFTR(inh)172. For further testing of the role of CFTR, metanephric cultures were prepared from mice with a targeted mutation of the Cftr gene. In contrast to kidneys from wild-type mice, those from Cftr -/- mice showed no evidence of tubular dilation in response to 8-Br-cAMP, indicating that CFTR Cl(-) channels are functional in embryonic kidneys and are required for cAMP-driven tubule expansion. A requirement for transepithelial Cl(-) transport was demonstrated by inhibiting the basolateral Na(+),K(+),2Cl(-) co-transporter with bumetanide, which effectively blocked all cAMP-stimulated tubular dilation. For determination of whether cystic dilation occurs to a greater extent in PKD kidneys in response to cAMP, Pkd1(m1Bei) -/- embryonic kidneys were treated with 8-Br-cAMP and were found to form rapidly CFTR and Na(+),K(+),2Cl(-) co-transporter-dependent cysts that were three- to six fold larger than those of wild-type kidneys. These results suggest that cAMP can stimulate fluid secretion early in renal tubule development during the time when renal cysts first appear in PKD kidneys and that PKD-deficient renal tubules are predisposed to abnormally increased cyst expansion in response to elevated levels of cAMP. PMID- 17108317 TI - beta(1) Receptors protect the renal afferent arteriole of angiotensin-infused rabbits from norepinephrine-induced oxidative stress. AB - Renal afferent arterioles (Aff) from angiotensin II (AngII)-infused rabbits have enhanced contractions to AngII that are normalized by tempol (superoxide dismutase mimetic), whereas contractions to norepinephrine (NE) are normal and unaffected by tempol. Tested was the hypothesis that beta-receptor stimulation with NE prevents enhanced reactivity and superoxide generation. Preconstricted Aff from AngII- or vehicle-infused rabbits were perfused at physiologic pressure. Aff from vehicle-infused rabbits had strong, endothelium-independent relaxations to dobutamine (beta(1)-receptor agonist; 78 +/- 6%; P < 0.0001; mean +/- SD) but only weak relaxations to salbutamol (beta(2)-receptor agonist; 13 +/- 3%; P < 0.05) or BRL-37,344 (beta(3)-receptor agonist; 14 +/- 3%; P < 0.05). Contractions to NE were similar in Aff from vehicle- and AngII-infused rabbits (-36 +/- 5 versus -34 +/- 3%; NS) and were unaffected by tempol (-32 +/- 4%; NS). In contrast, phenylephrine contractions (alpha(1) agonist) were enhanced in Aff from AngII-infused rabbits (-59 +/- 6 versus -46 +/- 4%; P < 0.05) and normalized by tempol. NE contractions in Aff from AngII-infused rabbits (-34 +/- 4%) were enhanced (P < 0.01) by propranolol (nonselective beta antagonist; -53 +/- 6%), CGP-20,712A (selective beta(1)-receptor antagonist; -61 +/- 9%), or Rp-cAMP (competitive inhibitor of cAMP; -56 +/- 4%); were normalized by tempol; but were unaffected by ICI-118,551 (selective beta(2)-receptor antagonist) or SR-59,230A (selective beta(3)-receptor antagonist). Superoxide generation in Aff from AngII infused rabbits that were assessed from ethidium:dihydroethidium was enhanced by addition of CGP-20,712A to NE but was normalized by tempol. Aff have robust alpha(1)-receptor contraction and beta(1)-receptor dilation. NE elicits beta(1) signaling via cAMP that moderates oxidative stress and contractions in Aff from AngII-infused rabbits. PMID- 17108318 TI - International differences in dialysis mortality reflect background general population atherosclerotic cardiovascular mortality. AB - Existing national, racial, and ethnic differences in dialysis patient mortality rates largely are unexplained. This study aimed to test the hypothesis that mortality rates related to atherosclerotic cardiovascular disease (ASCVD) in dialysis populations (DP) and in the background general populations (GP) are correlated. In a cross-sectional, multinational study, all-cause and ASCVD mortality rates were compared between GP and DP using the most recent data from the World Health Organization mortality database (67 countries; 1,571,852,000 population) and from national renal registries (26 countries; 623,900 population). Across GP of 67 countries (14,082,146 deaths), all-cause mortality rates (median 8.88 per 1000 population; range 1.93 to 15.40) were strongly related to ASCVD mortality rates (median 3.21; range 0.53 to 8.69), with Eastern European countries clustering in the upper and Southeast and East Asian countries in the lower rate ranges. Across DP (103,432 deaths), mortality rates from all causes (median 166.20; range 54.47 to 268.80) and from ASCVD (median 63.39 per 1000 population; range 21.52 to 162.40) were higher and strongly correlated. ASCVD mortality rates in DP and in the GP were significantly correlated; the relationship became even stronger after adjustment for age (R(2) = 0.56, P < 0.0001). A substantial portion of the variability in mortality rates that were observed across DP worldwide is attributable to the variability in background ASCVD mortality rates in the respective GP. Genetic and environmental factors may underlie these differences. PMID- 17108319 TI - Characterization of intron loss events in mammals. AB - The exon/intron structure of eukaryotic genes differs extensively across species, but the mechanisms and relative rates of intron loss and gain are still poorly understood. Here, we used whole-genome sequence alignments of human, mouse, rat, and dog to perform a genome-wide analysis of intron loss and gain events in >17,000 mammalian genes. We found no evidence for intron gain and 122 cases of intron loss, most of which occurred within the rodent lineage. The majority (68%) of the deleted introns were extremely small (<150 bp), significantly smaller than average. The intron losses occurred almost exclusively within highly expressed, housekeeping genes, supporting the hypothesis that intron loss is mediated via germline recombination of genomic DNA with intronless cDNA. This study constitutes the largest scale analysis for intron dynamics in vertebrates to date and allows us to confirm and extend several hypotheses previously based on much smaller samples. Our results in mammals show that intron gain has not been a factor in the evolution of gene structure during the past 95 Myr and has likely been restricted to more ancient history. PMID- 17108320 TI - Cdk5 Modulation of mitogen-activated protein kinase signaling regulates neuronal survival. AB - Cdk5, a cyclin-dependent kinase, is critical for neuronal development, neuronal migration, cortical lamination, and survival. Its survival role is based, in part, on "cross-talk" interactions with apoptotic and survival signaling pathways. Previously, we showed that Cdk5 phosphorylation of mitogen-activated protein kinase kinase (MEK)1 inhibits transient activation induced by nerve growth factor (NGF) in PC12 cells. To further explore the nature of this inhibition, we studied the kinetics of NGF activation of extracellular signal regulated kinase (Erk)1/2 in cortical neurons with or without roscovitine, an inhibitor of Cdk5. NGF alone induced an Erk1/2-transient activation that peaked in 15 min and declined rapidly to baseline. Roscovitine, alone or with NGF, reached peak Erk1/2 activation in 30 min that was sustained for 48 h. Moreover, the sustained Erk1/2 activation induced apoptosis in cortical neurons. Significantly, pharmacological application of the MEK1 inhibitor PD98095 to roscovitine-treated cortical neurons prevented apoptosis. These results were also confirmed by knocking down Cdk5 activity in cortical neurons with Cdk5 small interference RNA. Apoptosis was correlated with a significant shift of phosphorylated tau and neurofilaments from axons to neuronal cell bodies. These results suggest that survival of cortical neurons is also dependent on tight Cdk5 modulation of the mitogen-activated protein kinase signaling pathway. PMID- 17108321 TI - Mitochondria-associated yeast mRNAs and the biogenesis of molecular complexes. AB - The coherence of mitochondrial biogenesis relies on spatiotemporally coordinated associations of 800-1000 proteins mostly encoded in the nuclear genome. We report the development of new quantitative analyses to assess the role of local protein translation in the construction of molecular complexes. We used real-time PCR to determine the cellular location of 112 mRNAs involved in seven mitochondrial complexes. Five typical cases were examined by an improved FISH protocol. The proteins produced in the vicinity of mitochondria (MLR proteins) were, almost exclusively, of prokaryotic origin and are key elements of the core construction of the molecular complexes; the accessory proteins were translated on free cytoplasmic polysomes. These two classes of proteins correspond, at least as far as intermembrane space (IMS) proteins are concerned, to two different import pathways. Import of MLR proteins involves both TOM and TIM23 complexes whereas non-MLR proteins only interact with the TOM complex. Site-specific translation loci, both outside and inside mitochondria, may coordinate the construction of molecular complexes composed of both nuclearly and mitochondrially encoded subunits. PMID- 17108322 TI - Micromanipulation studies of chromatin fibers in Xenopus egg extracts reveal ATP dependent chromatin assembly dynamics. AB - We have studied assembly of chromatin using Xenopus egg extracts and single DNA molecules held at constant tension by using magnetic tweezers. In the absence of ATP, interphase extracts were able to assemble chromatin against DNA tensions of up to 3.5 piconewtons (pN). We observed force-induced disassembly and opening closing fluctuations, indicating our experiments were in mechanochemical equilibrium. Roughly 50-nm (150-base pair) lengthening events dominated force driven disassembly, suggesting that the assembled fibers are chiefly composed of nucleosomes. The ATP-depleted reaction was able to do mechanical work of 27 kcal/mol per 50 nm step, which provides an estimate of the free energy difference between core histone octamers on and off DNA. Addition of ATP led to highly dynamic behavior with time courses exhibiting processive runs of assembly and disassembly not observed in the ATP-depleted case. With ATP present, application of forces of 2 pN led to nearly complete fiber disassembly. Our study suggests that ATP hydrolysis plays a major role in nucleosome rearrangement and removal and that chromatin in vivo may be subject to highly dynamic assembly and disassembly processes that are modulated by DNA tension. PMID- 17108323 TI - Characterization of AtNUC-L1 reveals a central role of nucleolin in nucleolus organization and silencing of AtNUC-L2 gene in Arabidopsis. AB - Nucleolin is one of the most abundant protein in the nucleolus and is a multifunctional protein involved in different steps of ribosome biogenesis. In contrast to animals and yeast, the genome of the model plant Arabidopsis thaliana encodes two nucleolin-like proteins, AtNUC-L1 and AtNUC-L2. However, only the AtNUC-L1 gene is ubiquitously expressed in normal growth conditions. Disruption of this AtNUC-L1 gene leads to severe plant growth and development defects. AtNUC L1 is localized in the nucleolus, mainly in the dense fibrillar component. Absence of this protein in Atnuc-L1 plants induces nucleolar disorganization, nucleolus organizer region decondensation, and affects the accumulation levels of pre-rRNA precursors. Remarkably, in Atnuc-L1 plants the AtNUC-L2 gene is activated, suggesting that AtNUC-L2 might rescue, at least partially, the loss of AtNUC-L1. This work is the first description of a higher eukaryotic organism with a disrupted nucleolin-like gene and defines a new role for nucleolin in nucleolus structure and rDNA chromatin organization. PMID- 17108324 TI - Misfolded proteins traffic from the endoplasmic reticulum (ER) due to ER export signals. AB - Most misfolded secretory proteins remain in the endoplasmic reticulum (ER) and are degraded by ER-associated degradation (ERAD). However, some misfolded proteins exit the ER and traffic to the Golgi before degradation. Using model misfolded substrates, with or without defined ER exit signals, we found misfolded proteins can depart the ER by continuing to exhibit the functional export signals present in the corresponding correctly folded proteins. Anterograde transport of misfolded proteins utilizes the same machinery responsible for exporting correctly folded proteins. Passive ER retention, in which misfolded proteins fail to exit the ER due to the absence of exit signals or the inability to functionally present them, likely contributes to the retention of nonnative proteins in the ER. Intriguingly, compromising ERAD resulted in increased anterograde trafficking of a misfolded protein with an ER exit signal, suggesting that ERAD and ER exit machinery can compete for binding of misfolded proteins. Disabling ERAD did not result in transport of an ERAD substrate lacking an export signal. This is an important distinction for those seeking possible therapeutic approaches involving inactivating ERAD in anticipation of exporting a partially active protein. PMID- 17108325 TI - NuMA influences higher order chromatin organization in human mammary epithelium. AB - The coiled-coil protein NuMA is an important contributor to mitotic spindle formation and stabilization. A potential role for NuMA in nuclear organization or gene regulation is suggested by the observations that its pattern of nuclear distribution depends upon cell phenotype and that it interacts and/or colocalizes with transcription factors. To date, the precise contribution of NuMA to nuclear function remains unclear. Previously, we observed that antibody-induced alteration of NuMA distribution in growth-arrested and differentiated mammary epithelial structures (acini) in three-dimensional culture triggers the loss of acinar differentiation. Here, we show that in mammary epithelial cells, NuMA is present in both the nuclear matrix and chromatin compartments. Expression of a portion of the C terminus of NuMA that shares sequence similarity with the chromatin regulator HPC2 is sufficient to inhibit acinar differentiation and results in the redistribution of NuMA, chromatin markers acetyl-H4 and H4K20m, and regions of deoxyribonuclease I-sensitive chromatin compared with control cells. Short-term alteration of NuMA distribution with anti-NuMA C-terminus antibodies in live acinar cells indicates that changes in NuMA and chromatin organization precede loss of acinar differentiation. These findings suggest that NuMA has a role in mammary epithelial differentiation by influencing the organization of chromatin. PMID- 17108326 TI - A conserved dileucine motif mediates clathrin and AP-2-dependent endocytosis of the HIV-1 envelope protein. AB - During the assembly of enveloped viruses viral and cellular components essential for infectious particles must colocalize at specific membrane locations. For the human and simian immunodeficiency viruses (HIV and SIV), sorting of the viral envelope proteins (Env) to assembly sites is directed by trafficking signals located in the cytoplasmic domain of the transmembrane protein gp41 (TM). A membrane proximal conserved GYxxO motif mediates endocytosis through interaction with the clathrin adaptor AP-2. However, experiments with SIV(mac239) Env indicate the presence of additional signals. Here we show that a conserved C terminal dileucine in HIV(HxB2) also mediates endocytosis. Biochemical and morphological assays demonstrate that the C-terminal dileucine motif mediates internalization as efficiently as the GYxxO motif and that both must be removed to prevent Env internalization. RNAi experiments show that depletion of the clathrin adaptor AP-2 leads to increased plasma membrane expression of HIV Env and that this adaptor is required for efficient internalization mediated by both signals. The redundancy of conserved endocytosis signals and the role of the SIV(mac239) Env GYxxO motif in SIV pathogenesis, suggest that these motifs have functions in addition to endocytosis, possibly related to Env delivery to the site of viral assembly and/or incorporation into budding virions. PMID- 17108327 TI - The extracellular linker of pro-neuregulin-alpha2c is required for efficient sorting and juxtacrine function. AB - The neuregulins (NRGs) play important roles in animal physiology, and their disregulation has been linked to diseases such as cancer or schizophrenia. The NRGs may be produced as transmembrane proteins (proNRGs), even though they lack an N-terminal signal sequence. This raises the question of how NRGs are sorted to the plasma membrane. It is also unclear whether in their transmembrane state, the NRGs are biologically active. During studies aimed at solving these questions, we found that deletion of the extracellular juxtamembrane region termed the linker, decreased cell surface exposure of the mutant proNRG(DeltaLinker), and caused its entrapment at the cis-Golgi. We also found that cell surface-exposed transmembrane NRG forms retain biological activity. Thus, a mutant whose cleavage is impaired but is correctly sorted to the plasma membrane activated ErbB receptors in trans and also stimulated proliferation. Because the linker is implicated in surface sorting and the regulation of the cleavage of transmembrane NRGs, our data indicate that this region exerts multiple important roles in the physiology of NRGs. PMID- 17108328 TI - Internalization of large double-membrane intercellular vesicles by a clathrin dependent endocytic process. AB - Beyond its well-documented role in vesicle endocytosis, clathrin has also been implicated in the internalization of large particles such as viruses, pathogenic bacteria, and even latex beads. We have discovered an additional clathrin dependent endocytic process that results in the internalization of large, double membrane vesicles at lateral membranes of cells that are coupled by gap junctions (GJs). GJ channels bridge apposing cell membranes to mediate the direct transfer of electrical currents and signaling molecules from cell to cell. Here, we report that entire GJ plaques, clusters of GJ channels, can be internalized to form large, double-membrane vesicles previously termed annular gap junctions (AGJs). These internalized AGJ vesicles subdivide into smaller vesicles that are degraded by endo/lysosomal pathways. Mechanistic analyses revealed that clathrin-dependent endocytosis machinery-components, including clathrin itself, the alternative clathrin-adaptor Dab2, dynamin, myosin-VI, and actin are involved in the internalization, inward movement, and degradation of these large, intercellular double-membrane vesicles. These findings contribute to the understanding of clathrin's numerous emerging functions. PMID- 17108329 TI - SCFCdc4-mediated degradation of the Hac1p transcription factor regulates the unfolded protein response in Saccharomyces cerevisiae. AB - The Saccharomyces cerevisiae basic leucine zipper transcription factor Hac1p is synthesized in response to the accumulation of unfolded polypeptides in the lumen of the endoplasmic reticulum (ER), and it is responsible for up-regulation of approximately 5% of all yeast genes, including ER-resident chaperones and protein folding catalysts. Hac1p is one of the most short-lived yeast proteins, having a half-life of approximately 1.5 min. Here, we have shown that Hac1p harbors a functional PEST degron and that degradation of Hac1p by the proteasome involves the E2 ubiquitin-conjugating enzyme Ubc3/Cdc34p and the SCF(Cdc4) E3 complex. Consistent with the known nuclear localization of Cdc4p, rapid degradation of Hac1p requires the presence of a functional nuclear localization sequence, which we demonstrated to involve basic residues in the sequence (29)RKRAKTK(35). Two hybrid analysis demonstrated that the PEST-dependent interaction of Hac1p with Cdc4p requires Ser146 and Ser149. Turnover of Hac1p may be dependent on transcription because it is inhibited in cell mutants lacking Srb10 kinase, a component of the SRB/mediator module of the RNA polymerase II holoenzyme. Stabilization of Hac1p by point mutation or deletion, or as the consequence of defects in components of the degradation pathway, results in increased unfolded protein response element-dependent transcription and improved cell viability under ER stress conditions. PMID- 17108331 TI - Meiotic induction by heat stress in mouse oocytes: involvement of AMP-activated protein kinase and MAPK family members. AB - In this study, we examined the effect of heat pulsing on oocyte maturation and assessed the possible role of stress-activated enzymes during heat stress-induced meiotic maturation. Denuded oocytes from immature eCG-primed mice were pulsed for 30 min at increasing temperatures from 40 degrees C to 43 degrees C in dibutyryl cAMP-containing medium and were subsequently cultured at 37 degrees C for a total incubation time of 17-18 h. Oocytes exposed to 42 degrees C showed the greatest stimulation of maturation, with no effect at 43 degrees C. A heat pulse did not compromise progression to metaphase II as observed by polar body (PB) formation. The AMP-activated protein kinase (PRKA) inhibitors compound C and Ara-A each blocked the meiosis-stimulating effects of heat. Western blots showed that acetyl CoA carboxylase, an important substrate of PRKA, was phosphorylated in heat treated germinal vesicle-stage oocytes, indicating activation of PRKA before maturation. The mitogen-activated protein 2 kinase (MAP2K1) inhibitor PD98059 also prevented heat-induced maturation, but this effect was unrelated to MAPK1/3 activation, which was not observed until after germinal vesicle breakdown (GVB). Phosphorylated MAPK14 was not detected in the oocyte under any experimental condition, and only high concentrations of the MAPK14 inhibitor SB203580 blocked heat-stimulated maturation, suggesting that MAPK14 is not involved in meiotic induction. MAPK8/9 was activated by heat, and the MAPK8/9 inhibitor SP600125, but not JUN N-terminal kinase I, blocked heat-induced maturation. Heat treatment transiently suppressed GVB and PB formation in spontaneously maturing oocytes by a mechanism that is apparently different from its meiosis-inducing action. Collectively, these data show that an acute heat pulse stimulates GVB in meiotically arrested oocytes and suggest that this effect is mediated through the activation of PRKA. PMID- 17108330 TI - Pol I transcription and pre-rRNA processing are coordinated in a transcription dependent manner in mammalian cells. AB - Pre-rRNA synthesis and processing are key steps in ribosome biogenesis. Although recent evidence in yeast suggests that these two processes are coupled, the nature of their association is unclear. In this report, we analyze the coordination between rDNA transcription and pre-rRNA processing in mammalian cells. We found that pol I transcription factor UBF interacts with pre-rRNA processing factors as analyzed by immunoprecipitations, and the association depends on active rRNA synthesis. In addition, injections of plasmids containing the human rDNA promoter and varying lengths of 18S rDNA into HeLa nuclei show that pol I transcription machinery can be recruited to rDNA promoters regardless of the product that is transcribed, whereas subgroups of pre-rRNA processing factors are recruited to plasmids only when specific pre-rRNA fragments are produced. Our observations suggest a model for sequential recruitment of pol I transcription factors and pre-rRNA processing factors to elongating pre-rRNA on an as-needed basis rather than corecruitment to sites of active transcription. PMID- 17108333 TI - Production of F1 interspecies hybrid offspring with cryopreserved sperm from a live-bearing fish, the swordtail Xiphophorus helleri. AB - Despite study of sperm cryopreservation in more than 200 fish species, production of broods from cryopreserved sperm in live-bearing fish has not been demonstrated. This has not been due to a lack of effort, but instead is a result of the unique morphology, biology, and biochemistry of reproduction in viviparous fishes. For example, sperm of Xiphophorus helleri have a cylindrical nucleus, can swim for days after being activated, have glycolytic capabilities, and can reside in the female reproduction tract for months before fertilization. These traits are not found in fishes with external fertilization. The long-standing research use of the genus Xiphophorus has led to development of over 60 pedigreed lines among the 26 species maintained around the world. These species and lines serve as contemporary models in medical research, although they must be maintained as live populations. Previous attempts at establishing sperm cryopreservation protocols for Xiphophorus have not produced live young. To address this we have been studying the parameters surrounding cryobiology of Xiphophorus sperm and applying this information to an improved understanding of internal fertilization and reproduction. Here we report the first successful fertilization and offspring production by cryopreserved sperm in any live-bearing fish. This claim is supported by our use of artificial insemination between two species that yield distinct hybrid offspring to verify paternity via cryopreserved sperm. We provide a practical approach for preservation of valuable genetic resources from live bearing fish species, a group that is rapidly being lost due to destruction of native habitats. PMID- 17108332 TI - Low-density lipoprotein receptor-related protein 8 (LRP8) is upregulated in granulosa cells of bovine dominant follicle: molecular characterization and spatio-temporal expression studies. AB - The low-density lipoprotein (LDL) receptor-related protein 8 (LRP8) is a member of the LDL receptor family that participates in endocytosis and signal transduction. We cloned the full-length bovine LRP8 cDNA in granulosa cells (GC) of the dominant follicle (DF) as well as several LRP8 mRNA splicing variants, including a variant that contains a proline-rich cytoplasmic insert (A759-K817) that is involved in intracellular signaling. Expression of the A759-K817 variant was analyzed in the GC of follicles at different developmental stages: the small follicle (SF; 2-4 mm), the DF at Day 5 (D5) of the estrus cycle, ovulatory follicles (OF) 24 h after hCG injection, and corpora lutea (CL) at D5. RT-PCR analysis showed that expression was predominant in the GC of DF compared to other follicles and CL (P<0.0001), whereas the expression of other related receptors, such as LDLR and VLDLR, did not show differences. Temporal analyses of follicular walls from the OF following hCG treatment revealed a decrease in LRP8 mRNA expression starting 12 h post-hCG treatment (P<0.0001). LRP8 protein was exclusively localized to the GC, with higher levels in the DF than in the SF (P<0.05). RELN mRNA, which encodes an LRP8 ligand, was highly expressed in the theca of the DF as compared to the OF (P<0.004), whereas MAPK8IP1 mRNA, which encodes an LRP8 intracellular interacting partner, is expressed in the GC of the DF. These results demonstrate the differential expression patterns of LRP8, RELN, and MAPK8IP1 mRNAs during final follicular growth and ovulation, and suggest that a RELN/LRP8/MAPK8IP1 paracrine interaction regulates follicular growth. PMID- 17108334 TI - Differential regulation of colony stimulating factor 1 and macrophage migration inhibitory factor expression by inflammatory cytokines in term human decidua: implications for macrophage trafficking at the fetal-maternal interface. AB - Macrophages are a major component of the leukocyte population of human pregnant endometrium. Although several crucial functions have been ascribed to these cells, the mechanisms underlying macrophage trafficking in the placental bed are poorly understood. The aim of this study was to evaluate the in vivo expression of two potentially antagonistic macrophage-targeting chemokines, colony stimulating factor 1 (CSF1, also known as M-CSF) and macrophage migration inhibitory factor (MIF), in term decidua, and to examine the effects of the inflammatory cytokines tumor necrosis factor (TNF, also known as TNF alpha) and interleukin 1beta (IL1B) on CSF1 and MIF expression in cultured decidual cells. The expression of CSF1 and MIF in term decidua was evaluated by immunohistochemistry. Cultured decidual cells were primed with estradiol (E2) or with E2+medroxyprogesterone acetate (MPA), and then incubated with corresponding steroid(s) with or without TNF or IL1B. The levels of CSF1 and MIF protein and mRNA were assessed by ELISA and quantitative RT-PCR, respectively. Immunostaining for CSF1 and MIF was observed in term decidua. The levels of secreted CSF1 and MIF were similarly unchanged whether the decidual cells were incubated with E2 or with E2+MPA. The CSF1 levels significantly increased in cultures exposed to E2 or E2+MPA plus TNF or IL1B. In contrast, the MIF levels in TNF- and IL1B-treated cells were not changed significantly from the control cultures. The ELISA data were confirmed by quantitative RT-PCR analysis. These results indicate that CSF1 and MIF are involved in regulating macrophage trafficking at the fetal-maternal interface, and suggest a mechanism by which inflammatory cytokines influence pregnancy by regulating decidual macrophage infiltration. PMID- 17108335 TI - Platelet-derived growth factors and receptors in the rat corpus luteum: localization and identification of an effect on luteogenesis. AB - Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) play a vital role in regulating cell growth and angiogenesis. In this study, the expression of the family of PDGFs and PDGFRs in the ovarian corpus luteum were identified and characterized, and an effect of their activity on development of the corpus luteum revealed. Gonadotropin-stimulated immature rats were utilized as a model of induced ovulation, luteogenesis, and pseudopregnancy. Levels of ovarian mRNA for Pdgfb and Pdgfd, and their receptor, Pdgfrb, increased significantly as early as 4 h after human chorionic gonadotropin (hCG) injection in immature rats primed with equine chorionic gonadotropin (eCG). Gonadotropin regulation of Pdgfb expression was confirmed by in vitro promoter-reporter assays, which showed a 2- to 3-fold increase in Pdgfb promoter activity in response to luteinizing hormone (LH). Inhibition studies implicated protein kinase A, phosphatidylinositol 3 kinase and mitogen activated protein kinase signaling pathways in the LH-induced upregulation. In the corpus luteum, PDGFA, PDGFB, PDGFC, and PDGFRA were localized to a population of luteal parenchymal/steroidogenic cells. PDGFRB was expressed primarily in what appeared to be cells of the luteal microvasculature. Intraovarian injection of an inhibitor of PDGF receptor activity, the tyrphostin AG1295, prior to injection of hCG in eCG-primed immature rats resulted in a significant 21.86%+/-11.15% decrease in corpora lutea per treated ovary in comparison to the contralateral vehicle-injected control ovary. In addition, the treated ovary of 3 of 16 rats showed widespread hemorrhage throughout the entire ovary, indicating a possible role for PDGF receptor activity in maintenance of the ovarian vasculature. PMID- 17108336 TI - Premature chromosome condensation is not essential for nuclear reprogramming in bovine somatic cell nuclear transfer. AB - Premature chromosome condensation (PCC) was believed to promote nuclear reprogramming and to facilitate cloning by somatic cell nuclear transfer (NT) in mammalian species. However, it is still uncertain whether PCC is necessary for the successful reprogramming of an introduced donor nucleus in cattle. In the present study, fused NT embryos were subjected to immediate activation (IA, simultaneous fusion and activation), delayed activation (DA, activation applied 4 h postfusion), and IA with aged oocytes (IAA, activation at the same oocyte age as group DA). The morphologic changes, such as nuclear swelling, the occurrence of PCC, and microtubule/aster formation, were analyzed in detail by laser scanning confocal microscopy. When embryos were subjected to IA in both IA and IAA groups, the introduced nucleus gradually became swollen, and a pronuclear like structure formed within the oocyte, but PCC was not observed. In contrast, delaying embryo activation resulted in 46.5%-91.2% of NT embryos exhibiting PCC. This PCC was observed beginning at 4 h postcell fusion and was shown as one, two, or multiple chromosomal complexes. Subsequently, a diversity of pronuclear-like structures existed in NT embryos, characterized as single, double, and multiple nuclei. In the oocytes exhibiting PCC, the assembled spindle structure was observed to be an interactive mass, closely associated with condensed chromosomes, but no aster had formed. Regardless of whether they were subjected to IA, IAA, or DA treatments, if the oocytes contained pronuclear-like structures, either one or two asters were observed in proximity to the nuclei. A significantly higher rate of development to blastocysts was achieved in embryos that were immediately activated (IA, 59.1%; IAA, 40.7%) than in those for which activation was delayed (14.2%). The development rate was higher in group IA than in group IAA, but it was not significant (P = 0.089). Following embryo transfer, there was no statistically significant difference in the pregnancy rates (Day 70) between two of the groups (group IA, 11.7%, n = 94 vs. group DA, 12.3%, n = 130; P > 0.05) or live term development (group IA, 4.3% vs. group DA, 4.6%; P > 0.05). Our study has demonstrated that the IA of bovine NT embryos results in embryos with increased competence for preimplantational development. Moreover, PCC was shown to be unnecessary for the reprogramming of a transplanted somatic genome in a cattle oocyte. PMID- 17108337 TI - Cell-type localization of platelet-derived growth factors and receptors in the postnatal rat ovary and follicle. AB - Intraovarian growth factors play a significant role in the regulation of follicular selection and growth. In this study, the presence and localization of all members of the family of platelet-derived growth factors (PDGF) and receptors (PDGFR) were identified and characterized in the rat ovary. In addition, a role was identified for members of this family in contributing towards growth of preantral follicles. Real-time PCR revealed the presence of mRNA for all platelet derived growth factors (Pdgfa, Pdgfb, Pdgfc and Pdgfd) and receptors (Pdgfra and Pdgfrb) in the rat ovary from birth until 4 wk. In situ hybridization and immunohistochemistry were utilized to identify cell-type expression of PDGFs and PDGFRs in rat ovaries from birth until 4 wk. Shortly after birth, expression of PDGFRA and PDGFC was observed in and around oocyte clusters, and PDGFRB in stromal cells surrounding oocyte clusters. All members were identified in oocytes of primordial and primary follicles, and in cells of the theca layer of primordial to antral follicles. PDGFRA and PDGFA were also localized to some granulosa cells of secondary and antral follicles in ovaries from rats at Days 20 and 24. Thus, localization data suggest both theca-theca and theca-granulosa cell interactions of PDGFs and receptors. Preantral follicles cultured in vitro over 5 days in serum-free medium plus recombinant PDGFAA, PDGFAB, or PDGFBB increased in follicle diameter by 18.32%+/-2.18%, 17.72%+/-2.3%, and 17.6%+/-1.81%, respectively, representing significantly greater increases than for follicles incubated in serum-free medium alone (11%+/-1.57%), and suggesting a role for these growth factors in positively influencing early follicle growth. PMID- 17108338 TI - Images in clinical medicine. Minocycline-induced hyperpigmentation. PMID- 17108339 TI - Recalibrating Medicare payments for inpatient care. PMID- 17108340 TI - Violence and mental illness--how strong is the link? PMID- 17108341 TI - Murder or mercy? Hurricane Katrina and the need for disaster training. PMID- 17108342 TI - Normalization of hemoglobin level in patients with chronic kidney disease and anemia. AB - BACKGROUND: Whether correction of anemia in patients with stage 3 or 4 chronic kidney disease improves cardiovascular outcomes is not established. METHODS: We randomly assigned 603 patients with an estimated glomerular filtration rate (GFR) of 15.0 to 35.0 ml per minute per 1.73 m2 of body-surface area and mild-to moderate anemia (hemoglobin level, 11.0 to 12.5 g per deciliter) to a target hemoglobin value in the normal range (13.0 to 15.0 g per deciliter, group 1) or the subnormal range (10.5 to 11.5 g per deciliter, group 2). Subcutaneous erythropoietin (epoetin beta) was initiated at randomization (group 1) or only after the hemoglobin level fell below 10.5 g per deciliter (group 2). The primary end point was a composite of eight cardiovascular events; secondary end points included left ventricular mass index, quality-of-life scores, and the progression of chronic kidney disease. RESULTS: During the 3-year study, complete correction of anemia did not affect the likelihood of a first cardiovascular event (58 events in group 1 vs. 47 events in group 2; hazard ratio, 0.78; 95% confidence interval, 0.53 to 1.14; P=0.20). Left ventricular mass index remained stable in both groups. The mean estimated GFR was 24.9 ml per minute in group 1 and 24.2 ml per minute in group 2 at baseline and decreased by 3.6 and 3.1 ml per minute per year, respectively (P=0.40). Dialysis was required in more patients in group 1 than in group 2 (127 vs. 111, P=0.03). General health and physical function improved significantly (P=0.003 and P<0.001, respectively, in group 1, as compared with group 2). There was no significant difference in the combined incidence of adverse events between the two groups, but hypertensive episodes and headaches were more prevalent in group 1. CONCLUSIONS: In patients with chronic kidney disease, early complete correction of anemia does not reduce the risk of cardiovascular events. (ClinicalTrials.gov number, NCT00321919 [ClinicalTrials.gov].). PMID- 17108343 TI - Correction of anemia with epoetin alfa in chronic kidney disease. AB - BACKGROUND: Anemia, a common complication of chronic kidney disease, usually develops as a consequence of erythropoietin deficiency. Recombinant human erythropoietin (epoetin alfa) is indicated for the correction of anemia associated with this condition. However, the optimal level of hemoglobin correction is not defined. METHODS: In this open-label trial, we studied 1432 patients with chronic kidney disease, 715 of whom were randomly assigned to receive a dose of epoetin alfa targeted to achieve a hemoglobin level of 13.5 g per deciliter and 717 of whom were assigned to receive a dose targeted to achieve a level of 11.3 g per deciliter. The median study duration was 16 months. The primary end point was a composite of death, myocardial infarction, hospitalization for congestive heart failure (without renal replacement therapy), and stroke. RESULTS: A total of 222 composite events occurred: 125 events in the high-hemoglobin group, as compared with 97 events in the low-hemoglobin group (hazard ratio, 1.34; 95% confidence interval, 1.03 to 1.74; P=0.03). There were 65 deaths (29.3%), 101 hospitalizations for congestive heart failure (45.5%), 25 myocardial infarctions (11.3%), and 23 strokes (10.4%). Seven patients (3.2%) were hospitalized for congestive heart failure and myocardial infarction combined, and one patient (0.5%) died after having a stroke. Improvements in the quality of life were similar in the two groups. More patients in the high hemoglobin group had at least one serious adverse event. CONCLUSIONS: The use of a target hemoglobin level of 13.5 g per deciliter (as compared with 11.3 g per deciliter) was associated with increased risk and no incremental improvement in the quality of life. (ClinicalTrials.gov number, NCT00211120 [ClinicalTrials.gov].). PMID- 17108344 TI - Clinical practice. Acute bronchitis. PMID- 17108345 TI - Images in clinical medicine. Fibromuscular dysplasia of the renal artery. PMID- 17108346 TI - Case records of the Massachusetts General Hospital. Case 35-2006. A newborn boy with hypotonia. PMID- 17108347 TI - Correction of anemia--payoffs and problems. PMID- 17108348 TI - Therapy for methicillin-resistant Staphylococcus aureus. PMID- 17108349 TI - Aneurysm syndromes and TGF-beta receptor mutations. PMID- 17108350 TI - Alendronate or alfacalcidol in glucocorticoid-induced osteoporosis. PMID- 17108351 TI - More on life-threatening asthma and salmeterol. PMID- 17108352 TI - Atazanavir urolithiasis. PMID- 17108353 TI - DDBJ working on evaluation and classification of bacterial genes in INSDC. AB - DNA Data Bank of Japan (DDBJ) (http://www.ddbj.nig.ac.jp) newly collected and released 12,927,184 entries or 13,787,688,598 bases in the period from July 2005 to June 2006. The released data contain honeybee expressed sequence tags (ESTs), re-examined and re-annotated complete genome data of Escherichia coli K-12 W3110, medaka WGS and human MGA. We also systematically evaluated and classified the genes in the complete bacterial genomes submitted to the International Nucleotide Sequence Database Collaboration (INSDC, http://insdc.org) that is composed of DDBJ, EMBL Bank and GenBank. The examination and classification selected 557,000 genes as reliable ones among all the bacterial genes predicted by us. PMID- 17108354 TI - miRGen: a database for the study of animal microRNA genomic organization and function. AB - miRGen is an integrated database of (i) positional relationships between animal miRNAs and genomic annotation sets and (ii) animal miRNA targets according to combinations of widely used target prediction programs. A major goal of the database is the study of the relationship between miRNA genomic organization and miRNA function. This is made possible by three integrated and user friendly interfaces. The Genomics interface allows the user to explore where whole-genome collections of miRNAs are located with respect to UCSC genome browser annotation sets such as Known Genes, Refseq Genes, Genscan predicted genes, CpG islands and pseudogenes. These miRNAs are connected through the Targets interface to their experimentally supported target genes from TarBase, as well as computationally predicted target genes from optimized intersections and unions of several widely used mammalian target prediction programs. Finally, the Clusters interface provides predicted miRNA clusters at any given inter-miRNA distance and provides specific functional information on the targets of miRNAs within each cluster. All of these unique features of miRGen are designed to facilitate investigations into miRNA genomic organization, co-transcription and targeting. miRGen can be freely accessed at http://www.diana.pcbi.upenn.edu/miRGen. PMID- 17108355 TI - The ASAP II database: analysis and comparative genomics of alternative splicing in 15 animal species. AB - We have greatly expanded the Alternative Splicing Annotation Project (ASAP) database: (i) its human alternative splicing data are expanded approximately 3 fold over the previous ASAP database, to nearly 90,000 distinct alternative splicing events; (ii) it now provides genome-wide alternative splicing analyses for 15 vertebrate, insect and other animal species; (iii) it provides comprehensive comparative genomics information for comparing alternative splicing and splice site conservation across 17 aligned genomes, based on UCSC multigenome alignments; (iv) it provides an approximately 2- to 3-fold expansion in detection of tissue-specific alternative splicing events, and of cancer versus normal specific alternative splicing events. We have also constructed a novel database linking orthologous exons and orthologous introns between genomes, based on multigenome alignment of 17 animal species. It can be a valuable resource for studies of gene structure evolution. ASAP II provides a new web interface enabling more detailed exploration of the data, and integrating comparative genomics information with alternative splicing data. We provide a set of tools for advanced data-mining of ASAP II with Pygr (the Python Graph Database Framework for Bioinformatics) including powerful features such as graph query, multigenome alignment query, etc. ASAP II is available at http://www.bioinformatics.ucla.edu/ASAP2. PMID- 17108356 TI - Stimulation of MCM helicase activity by a Cdc6 protein in the archaeon Thermoplasma acidophilum. AB - Replicative DNA helicases are ring-shaped hexamers that play an essential role in chromosomal DNA replication. They unwind the two strands of the duplex DNA and provide the single-stranded (ss) DNA substrate for the polymerase. The minichromosome maintenance (MCM) proteins are thought to function as the replicative helicases in eukarya and archaea. The proteins of only a few archaeal organisms have been studied and revealed that although all have similar amino acid sequences and overall structures they differ in their biochemical properties. In this report the biochemical properties of the MCM protein from the archaeon Thermoplasma acidophilum is described. The enzyme has weak helicase activity on a substrate containing only a 3'-ssDNA overhang region and the protein requires a forked DNA structure for efficient helicase activity. It was also found that the helicase activity is stimulated by one of the two T.acidophilum Cdc6 homologues. This is an interesting observation as it is in sharp contrast to observations made with MCM and Cdc6 homologues from other archaea in which the helicase activity is inhibited when bound to Cdc6. PMID- 17108357 TI - The HIV positive selection mutation database. AB - The HIV positive selection mutation database is a large-scale database available at http://www.bioinformatics.ucla.edu/HIV/ that provides detailed selection pressure maps of HIV protease and reverse transcriptase, both of which are molecular targets of antiretroviral therapy. This database makes available for the first time a very large HIV sequence dataset (sequences from approximately 50 000 clinical AIDS samples, generously contributed by Specialty Laboratories, Inc.), which makes possible high-resolution selection pressure mapping. It provides information about not only the selection pressure on individual sites but also how selection pressure at one site is affected by mutations on other sites. It also includes datasets from other public databases, namely the Stanford HIV database [S. Y. Rhee, M. J. Gonzales, R. Kantor, B. J. Betts, J. Ravela and R. W. Shafer (2003) Nucleic Acids Res., 31, 298-303]. Comparison between these datasets in the database enables cross-validation with independent datasets and also specific evaluation of the effect of drug treatment. PMID- 17108358 TI - A microfluidic-FCS platform for investigation on the dissociation of Sp1-DNA complex by doxorubicin. AB - The transcription factor (TF) Sp1 is a well-known RNA polymerase II transcription activator that binds to GC-rich recognition sites in a number of essential cellular and viral promoters. In addition, direct interference of Sp1 binding to DNA cognate sites using DNA-interacting compounds may provide promising therapies for suppression of cancer progression and viral replication. In this study, we present a rapid, sensitive and cost-effective evaluation of a GC intercalative drug, doxorubicin (DOX), in dissociating the Sp1-DNA complex using fluorescence correlation spectroscopy (FCS) in a microfluidic system. FCS allows assay miniaturization without compromising sensitivity, making it an ideal analytical method for integration of binding assays into high-throughput, microfluidic platforms. A polydimethylsiloxane (PDMS)-based microfluidic chip with a mixing network is used to achieve specific drug concentrations for drug titration experiments. Using FCS measurements, the IC50 of DOX on the dissociation of Sp1 DNA complex is estimated to be 0.55 microM, which is comparable to that measured by the electrophoretic mobility shift assay (EMSA). However, completion of one drug titration experiment on the proposed microfluidic-FCS platform is accomplished using only picograms of protein and DNA samples and less than 1 h total assay time, demonstrating vast improvements over traditional ensemble techniques. PMID- 17108359 TI - The telomerase-recruitment domain of the telomere binding protein Cdc13 is regulated by Mec1p/Tel1p-dependent phosphorylation. AB - The DNA damage-responsive protein kinases ATM and ATR phosphorylate SQ/TQ motifs that lie in clusters in most of their in vivo targets. Budding yeast Cdc13p contains two clusters of SQ/TQ motifs, suggesting that it might be a target of Mec1p/Tel1p (yeast ATR/ATM). Here we demonstrated that the telomerase recruitment domain of Cdc13p is phosphorylated by Mec1p and Tel1p. Gel analysis showed that Cdc13p contains a Mec1/Tel1-dependent post-translational modification. Using an immunoprecipitate (IP)-kinase assay, we showed that Mec1p phosphorylates Cdc13p on serine 225, 249, 255 and 306, and Tel1p phosphorylates Cdc13p on serine 225, 249 and 255 in vitro. Phenotypic analysis in vivo revealed that the mutations in the Cdc13p SQ motifs phosphorylated by Mec1p and Tel1p caused multiple telomere and growth defects. In addition, normal telomere length and growth could be restored by expressing a Cdc13-Est1p hybrid protein. These results demonstrate the telomerase recruitment domain of Cdc13p as an important new telomere-specific target of Mec1p/Tel1p. PMID- 17108360 TI - The Online Bioinformatics Resources Collection at the University of Pittsburgh Health Sciences Library System--a one-stop gateway to online bioinformatics databases and software tools. AB - To bridge the gap between the rising information needs of biological and medical researchers and the rapidly growing number of online bioinformatics resources, we have created the Online Bioinformatics Resources Collection (OBRC) at the Health Sciences Library System (HSLS) at the University of Pittsburgh. The OBRC, containing 1542 major online bioinformatics databases and software tools, was constructed using the HSLS content management system built on the Zope Web application server. To enhance the output of search results, we further implemented the Vivisimo Clustering Engine, which automatically organizes the search results into categories created dynamically based on the textual information of the retrieved records. As the largest online collection of its kind and the only one with advanced search results clustering, OBRC is aimed at becoming a one-stop guided information gateway to the major bioinformatics databases and software tools on the Web. OBRC is available at the University of Pittsburgh's HSLS Web site (http://www.hsls.pitt.edu/guides/genetics/obrc). PMID- 17108361 TI - eSLDB: eukaryotic subcellular localization database. AB - Eukaryotic Subcellular Localization DataBase collects the annotations of subcellular localization of eukaryotic proteomes. So far five proteomes have been processed and stored: Homo sapiens, Mus musculus, Caenorhabditis elegans, Saccharomyces cerevisiae and Arabidopsis thaliana. For each sequence, the database lists localization obtained adopting three different approaches: (i) experimentally determined (when available); (ii) homology-based (when possible); and (iii) predicted. The latter is computed with a suite of machine learning based methods, developed in house. All the data are available at our website and can be searched by sequence, by protein code and/or by protein description. Furthermore, a more complex search can be performed combining different search fields and keys. All the data contained in the database can be freely downloaded in flat file format. The database is available at http://gpcr.biocomp.unibo.it/esldb/. PMID- 17108362 TI - High-density genotyping and functional SNP localization in the CETP gene. AB - The cholesteryl ester transfer protein gene (CETP) has been the subject of hundreds of genetic analyses that typically focus on a small number of polymorphisms within a single ethnic group. Furthermore, the extent of DNA beyond the transcribed sequence from which single nucleotide polymorphisms (SNPs) may influence CETP expression has not been well defined. To better understand the role of natural variation in modulating CETP and high density lipoprotein cholesterol (HDL-C) levels, dense genotyping of CETP and regions up to 15 kb on either side of the gene was carried out on >2,000 individuals. A complex, nonlinear set of linkage disequilibrium bins was found, with many bins interspersed along the DNA sequence and spread over large regions of the gene. Bins assigned based on large numbers of individuals matched the small subset of SNPs that had been assigned to bins previously with a small number of individuals. Associations of known functional SNPs with HDL-C were found, but there were suggestions that there are additional functional SNPs not characterized previously. Narrowing of the set of likely functional SNPs was accomplished by comparing associations observed in different ethnic groups. The promoter SNP most highly associated with HDL-C that is likely to be functional, position -4,502, alters a consensus transcription factor binding site. PMID- 17108363 TI - Efficient generation of transgenic rats through the male germline using lentiviral transduction and transplantation of spermatogonial stem cells. AB - Spermatozoa produced from spermatogonial stem cells (SSCs) are the vehicle by which genes of a male are passed to the next generation. A single SSC has the ability to self-renew and produce thousands of spermatozoa; therefore, it is an ideal target for genetic modification to efficiently generate transgenic animals in mammalian species. Rats are an important model organism for biological research; however, gene function studies have been difficult because of a limited ability to generate transgenic animals. Transgenic rat production through SSCs offers a means to overcome this obstacle. Because SSCs divide slowly both in vivo and in vitro, lentiviral vectors may be an ideal method for introducing stable genetic modification. Using a lentiviral vector, an enhanced green fluorescent protein (eGFP) transgene was introduced into the genome of cultured rat SSCs, which were microinjected into testes of immunodeficient mice to assess transduction efficiency. Approximately 40% of rat SSCs exposed to the lentiviral vector overnight carried the eGFP transgene and generated colonies of spermatogenesis. When transduced SSCs were transplanted into recipient rat testes, in which endogenous germ cells had been decreased but not eliminated by busulfan treatment, approximately 6% of offspring were transgenic. The transgene was stably integrated into the donor SSC genome and transmitted to and expressed by progeny in subsequent generations. Thus, lentiviral transduction of SSCs followed by transplantation is an effective means for generating transgenic rats through the male germline, and this approach may be applicable to other species in which existing methods are inadequate or not applicable. PMID- 17108364 TI - Characterizing the glycocalyx of poultry spermatozoa: I. Identification and distribution of carbohydrate residues using flow cytometry and epifluorescence microscopy. AB - The aim of the present work was to use a battery of lectins to 1) delineate the carbohydrate content of sperm glycocalyx in the turkey and chicken using flow cytometry analysis, and 2) evaluate the distribution of existing sugars over the sperm plasma membrane surface with epifluorescent microscopy. Carbohydrate groups (corresponding lectins) that were investigated included galactose (GS-I, Jacalin, RCA-I, PNA), glucose and/or mannose (Con A, PSA, GNA), N-acetyl-glucosamine (GS II, s-WGA, STA), N-acetyl-galactosamine (SBA, WFA), fucose (Lotus, UEA-I), sialic acid (LFA, LPA), and N-acetyl-lactosamine (ECA). Spermatozoa were assessed before and after treatment with neuraminidase to remove sialic acid. Mean fluorescence intensity (MnFI) was used as indicator of lectin binding for flow cytometry analysis. Nontreated spermatozoa from both species showed high MnFI when incubated with RCA-I, Con A, LFA, and LPA, as did chicken spermatozoa incubated with s-WGA. Neuraminidase treatment increased the MnFI for most lectins except LFA and LPA, as expected. Differences in MnFI between species included higher values for s-WGA and ECA in chicken spermatozoa and for WFA in turkey spermatozoa. Microscopy revealed segregation of some sugar residues into membrane specific domains; however, the 2 staining techniques (cell suspension vs fixed preparation) differed in identifying lectin binding patterns, with fixed preparations yielding a high degree of nonspecific binding. We conclude that 1) the glycocalyx of turkey and chicken spermatozoa contains a diversity of carbohydrate groups, 2) these residues are extensively masked by sialic acid, 3) the glycocalyx composition is species-specific, and 4) some glycoconjugates appear to be segregated into membrane-specific domains. Characterization of the poultry sperm glycocalyx is the first step in identifying the physiological impact of semen storage on sperm function. PMID- 17108365 TI - Random multispace quantization as an analytic mechanism for BioHashing of biometric and random identity inputs. AB - Biometric analysis for identity verification is becoming a widespread reality. Such implementations necessitate large-scale capture and storage of biometric data, which raises serious issues in terms of data privacy and (if such data is compromised) identity theft. These problems stem from the essential permanence of biometric data, which (unlike secret passwords or physical tokens) cannot be refreshed or reissued if compromised. Our previously presented biometric-hash framework prescribes the integration of external (password or token-derived) randomness with user-specific biometrics, resulting in bitstring outputs with security characteristics (i.e., noninvertibility) comparable to cryptographic ciphers or hashes. The resultant BioHashes are hence cancellable, i.e., straightforwardly revoked and reissued (via refreshed password or reissued token) if compromised. BioHashing furthermore enhances recognition effectiveness, which is explained in this paper as arising from the Random Multispace Quantization (RMQ) of biometric and external random inputs. PMID- 17108366 TI - Validating a biometric authentication system: sample size requirements. AB - Authentication systems based on biometric features (e.g., fingerprint impressions, iris scans, human face images, etc.) are increasingly gaining widespread use and popularity. Often, vendors and owners of these commercial biometric systems claim impressive performance that is estimated based on some proprietary data. In such situations, there is a need to independently validate the claimed performance levels. System performance is typically evaluated by collecting biometric templates from n different subjects, and for convenience, acquiring multiple instances of the biometric for each of the n subjects. Very little work has been done in 1) constructing confidence regions based on the ROC curve for validating the claimed performance levels and 2) determining the required number of biometric samples needed to establish confidence regions of prespecified width for the ROC curve. To simplify the analysis that address these two problems, several previous studies have assumed that multiple acquisitions of the biometric entity are statistically independent. This assumption is too restrictive and is generally not valid. We have developed a validation technique based on multivariate copula models for correlated biometric acquisitions. Based on the same model, we also determine the minimum number of samples required to achieve confidence bands of desired width for the ROC curve. We illustrate the estimation of the confidence bands as well as the required number of biometric samples using a fingerprint matching system that is applied on samples collected from a small population. PMID- 17108367 TI - Recovering facial shape using a statistical model of surface normal direction. AB - In this paper, we show how a statistical model of facial shape can be embedded within a shape-from-shading algorithm. We describe how facial shape can be captured using a statistical model of variations in surface normal direction. To construct this model, we make use of the azimuthal equidistant projection to map the distribution of surface normals from the polar representation on a unit sphere to Cartesian points on a local tangent plane. The distribution of surface normal directions is captured using the covariance matrix for the projected point positions. The eigenvectors of the covariance matrix define the modes of shape variation in the fields of transformed surface normals. We show how this model can be trained using surface normal data acquired from range images and how to fit the model to intensity images of faces using constraints on the surface normal direction provided by Lambert's law. We demonstrate that the combination of a global statistical constraint and local irradiance constraint yields an efficient and accurate approach to facial shape recovery and is capable of recovering fine local surface details. We assess the accuracy of the technique on a variety of images with ground truth and real-world images. PMID- 17108368 TI - MILES: multiple-instance learning via embedded instance selection. AB - Multiple-instance problems arise from the situations where training class labels are attached to sets of samples (named bags), instead of individual samples within each bag (called instances). Most previous multiple-instance learning (MIL) algorithms are developed based on the assumption that a bag is positive if and only if at least one of its instances is positive. Although the assumption works well in a drug activity prediction problem, it is rather restrictive for other applications, especially those in the computer vision area. We propose a learning method, MILES (Multiple-Instance Learning via Embedded instance Selection), which converts the multiple-instance learning problem to a standard supervised learning problem that does not impose the assumption relating instance labels to bag labels. MILES maps each bag into a feature space defined by the instances in the training bags via an instance similarity measure. This feature mapping often provides a large number of redundant or irrelevant features. Hence, 1-norm SVM is applied to select important features as well as construct classifiers simultaneously. We have performed extensive experiments. In comparison with other methods, MILES demonstrates competitive classification accuracy, high computation efficiency, and robustness to labeling uncertainty. PMID- 17108369 TI - Bayesian Gaussian process classification with the EM-EP algorithm. AB - Gaussian process classifiers (GPCs) are Bayesian probabilistic kernel classifiers. In GPCs, the probability of belonging to a certain class at an input location is monotonically related to the value of some latent function at that location. Starting from a Gaussian process prior over this latent function, data are used to infer both the posterior over the latent function and the values of hyperparameters to determine various aspects of the function. Recently, the expectation propagation (EP) approach has been proposed to infer the posterior over the latent function. Based on this work, we present an approximate EM algorithm, the EM-EP algorithm, to learn both the latent function and the hyperparameters. This algorithm is found to converge in practice and provides an efficient Bayesian framework for learning hyperparameters of the kernel. A multiclass extension of the EM-EP algorithm for GPCs is also derived. In the experimental results, the EM-EP algorithms are as good or better than other methods for GPCs or Support Vector Machines (SVMs) with cross-validation. PMID- 17108370 TI - MCMC data association and sparse factorization updating for real time multitarget tracking with merged and multiple measurements. AB - In several multitarget tracking applications, a target may return more than one measurement per target and interacting targets may return multiple merged measurements between targets. Existing algorithms for tracking and data association, initially applied to radar tracking, do not adequately address these types of measurements. Here, we introduce a probabilistic model for interacting targets that addresses both types of measurements simultaneously. We provide an algorithm for approximate inference in this model using a Markov chain Monte Carlo (MCMC)-based auxiliary variable particle filter. We Rao-Blackwellize the Markov chain to eliminate sampling over the continuous state space of the targets. A major contribution of this work is the use of sparse least squares updating and downdating techniques, which significantly reduce the computational cost per iteration of the Markov chain. Also, when combined with a simple heuristic, they enable the algorithm to correctly focus computation on interacting targets. We include experimental results on a challenging simulation sequence. We test the accuracy of the algorithm using two sensor modalities, video, and laser range data. We also show the algorithm exhibits real time performance on a conventional PC. PMID- 17108371 TI - On the distribution of saliency. AB - Detecting salient structures is a basic task in perceptual organization. Saliency algorithms typically mark edge-points with some saliency measure, which grows with the length and smoothness of the curve on which these edge-points lie. Here, we propose a modified saliency estimation mechanism that is based on probabilistically specified grouping cues and on curve length distributions. In this framework, the Shashua and Ullman saliency mechanism may be interpreted as a process for detecting the curve with maximal expected length. Generalized types of saliency naturally follow. We propose several specific generalizations (e.g., gray-level-based saliency) and rigorously derive the limitations on generalized saliency types. We then carry out a probabilistic analysis of expected length saliencies. Using ergodicity and asymptotic analysis, we derive the saliency distributions associated with the main curves and with the rest of the image. We then extend this analysis to finite-length curves. Using the derived distributions, we derive the optimal threshold on the saliency for discriminating between figure and background and bound the saliency-based figure-from-ground performance. PMID- 17108372 TI - Shape representation and classification using the poisson equation. AB - We present a novel approach that allows us to reliably compute many useful properties of a silhouette. Our approach assigns, for every internal point of the silhouette, a value reflecting the mean time required for a random walk beginning at the point to hit the boundaries. This function can be computed by solving Poisson's equation, with the silhouette contours providing boundary conditions. We show how this function can be used to reliably extract various shape properties including part structure and rough skeleton, local orientation and aspect ratio of different parts, and convex and concave sections of the boundaries. In addition to this, we discuss properties of the solution and show how to efficiently compute this solution using multigrid algorithms. We demonstrate the utility of the extracted properties by using them for shape classification and retrieval. PMID- 17108373 TI - Dynamic shape and appearance models. AB - We propose a model of the joint variation of shape and appearance of portions of an image sequence. The model is conditionally linear, and can be thought of as an extension of active appearance models to exploit the temporal correlation of adjacent image frames. Inference of the model parameters can be performed efficiently using established numerical optimization techniques borrowed from finite-element analysis and system identification techniques. PMID- 17108374 TI - Symbol recognition with kernel density matching. AB - We propose a novel approach to similarity assessment for graphic symbols. Symbols are represented as 2D kernel densities and their similarity is measured by the Kullback-Leibler divergence. Symbol orientation is found by gradient-based angle searching or independent component analysis. Experimental results show the outstanding performance of this approach in various situations. PMID- 17108375 TI - Robust pose estimation from a planar target. AB - In theory, the pose of a calibrated camera can be uniquely determined from a minimum of four coplanar but noncollinear points. In practice, there are many applications of camera pose tracking from planar targets and there is also a number of recent pose estimation algorithms which perform this task in real-time, but all of these algorithms suffer from pose ambiguities. This paper investigates the pose ambiguity for planar targets viewed by a perspective camera. We show that pose ambiguities--two distinct local minima of the according error function- exist even for cases with wide angle lenses and close range targets. We give a comprehensive interpretation of the two minima and derive an analytical solution that locates the second minimum. Based on this solution, we develop a new algorithm for unique and robust pose estimation from a planar target. In the experimental evaluation, this algorithm outperforms four state-of-the-art pose estimation algorithms. PMID- 17108376 TI - Geometric interpretations of the relation between the image of the absolute conic and sphere images. AB - A spherical object has been introduced into camera calibration for several years through utilizing the properties of an image conic, which is the projection of the occluding contour of a sphere in the perspective image. However, in literature, only an algebraic interpretation was presented for the relation between the image of the absolute conic and sphere images. In this paper, we propose two geometric interpretations of this relation and two novel camera calibration methods using sphere images are derived from these geometric interpretations. PMID- 17108377 TI - Face description with local binary patterns: application to face recognition. AB - This paper presents a novel and efficient facial image representation based on local binary pattern (LBP) texture features. The face image is divided into several regions from which the LBP feature distributions are extracted and concatenated into an enhanced feature vector to be used as a face descriptor. The performance of the proposed method is assessed in the face recognition problem under different challenges. Other applications and several extensions are also discussed. PMID- 17108378 TI - A luminance- and contrast-invariant edge-similarity measure. AB - A novel similarity measure for edge-detection that is robust to varying luminance and contrast is presented. It incorporates a regularization term and employs directional FIR edge filters with hyperbolic tangent profiles to ensure improved noise performance and edge localization compared to classical methods. PMID- 17108379 TI - Local or global minima: flexible dual-front active contours. AB - Most variational active contour models are designed to find local minima of data dependent energy functionals with the hope that reasonable initial placement of the active contour will drive it toward a "desirable" local minimum as opposed to an undesirable configuration due to noise or complex image structure. As such, there has been much research into the design of complex region-based energy functionals that are less likely to yield undesirable local minima when compared to simpler edge-based energy functionals whose sensitivity to noise and texture is significantly worse. Unfortunately, most of these more "robust" region-based energy functionals are applicable to a much narrower class of imagery compared to typical edge-based energies due to stronger global assumptions about the underlying image data. Devising new implementation algorithms for active contours that attempt to capture more global minimizers of already proposed image-based energies would allow us to choose an energy that makes sense for a particular class of energy without concern over its sensitivity to local minima. Such implementations have been proposed for capturing global minima. However, sometimes the completely-global minimum is just as undesirable as a minimum that is too local. In this paper, we propose a novel, fast, and flexible dual front implementation of active contours, motivated by minimal path techniques and utilizing fast sweeping algorithms, which is easily manipulated to yield minima with variable "degrees" of localness and globalness. By simply adjusting the size of active regions, the ability to gracefully move from capturing minima that are more local (according to the initial placement of the active contour/surface) to minima that are more global allows this model to more easily obtain "desirable" minimizers (which often are neither the most local nor the most global). Experiments on various 2D and 3D images and comparisons with some active contour models and region-growing methods are also given to illustrate the properties of this model and its performance in a variety of segmentation applications. PMID- 17108380 TI - Pores and ridges: high-resolution fingerprint matching using level 3 features. AB - Fingerprint friction ridge details are generally described in a hierarchical order at three different levels, namely, Level 1 (pattern), Level 2 (minutia points), and Level 3 (pores and ridge contours). Although latent print examiners frequently take advantage of Level 3 features to assist in identification, Automated Fingerprint Identification Systems (AFIS) currently rely only on Level 1 and Level 2 features. In fact, the Federal Bureau of Investigation's (FBI) standard of fingerprint resolution for AFIS is 500 pixels per inch (ppi), which is inadequate for capturing Level 3 features, such as pores. With the advances in fingerprint sensing technology, many sensors are now equipped with dual resolution (500 ppi/1,000 ppi) scanning capability. However, increasing the scan resolution alone does not necessarily provide any performance improvement in fingerprint matching, unless an extended feature set is utilized. As a result, a systematic study to determine how much performance gain one can achieve by introducing Level 3 features in AFIS is highly desired. We propose a hierarchical matching system that utilizes features at all the three levels extracted from 1,000 ppi fingerprint scans. Level 3 features, including pores and ridge contours, are automatically extracted using Gabor filters and wavelet transform and are locally matched using the Iterative Closest Point (ICP) algorithm. Our experiments show that Level 3 features carry significant discriminatory information. There is a relative reduction of 20 percent in the equal error rate (EER) of the matching system when Level 3 features are employed in combination with Level 1 and 2 features. This significant performance gain is consistently observed across various quality fingerprint images. PMID- 17108381 TI - Joint optimization of word alignment and epenthesis generation for Chinese to Taiwanese sign synthesis. AB - This work proposes a novel approach to translate Chinese to Taiwanese sign language and to synthesize sign videos. An aligned bilingual corpus of Chinese and Taiwanese Sign Language (TSL) with linguistic and signing information is also presented for sign language translation. A two-pass alignment in syntax level and phrase level is developed to obtain the optimal alignment between Chinese sentences and Taiwanese sign sequences. For sign video synthesis, a scoring function is presented to develop motion transition-balanced sign videos with rich combinations of intersign transitions. Finally, the maximum a posteriori (MAP) algorithm is employed for sign video synthesis based on joint optimization of two pass word alignment and intersign epenthesis generation. Several experiments are conducted in an educational environment to evaluate the performance on the comprehension of sign expression. The proposed approach outperforms the IBM Model 2 in sign language translation. Moreover, deaf students perceived sign videos generated by the proposed method to be satisfactory. PMID- 17108382 TI - Graph embedding and extensions: a general framework for dimensionality reduction. AB - Over the past few decades, a large family of algorithms - supervised or unsupervised; stemming from statistics or geometry theory - has been designed to provide different solutions to the problem of dimensionality reduction. Despite the different motivations of these algorithms, we present in this paper a general formulation known as graph embedding to unify them within a common framework. In graph embedding, each algorithm can be considered as the direct graph embedding or its linear/kernel/tensor extension of a specific intrinsic graph that describes certain desired statistical or geometric properties of a data set, with constraints from scale normalization or a penalty graph that characterizes a statistical or geometric property that should be avoided. Furthermore, the graph embedding framework can be used as a general platform for developing new dimensionality reduction algorithms. By utilizing this framework as a tool, we propose a new supervised dimensionality reduction algorithm called Marginal Fisher Analysis in which the intrinsic graph characterizes the intraclass compactness and connects each data point with its neighboring points of the same class, while the penalty graph connects the marginal points and characterizes the interclass separability. We show that MFA effectively overcomes the limitations of the traditional Linear Discriminant Analysis algorithm due to data distribution assumptions and available projection directions. Real face recognition experiments show the superiority of our proposed MFA in comparison to LDA, also for corresponding kernel and tensor extensions. PMID- 17108383 TI - Spatio-temporal context for robust multitarget tracking. AB - In multitarget tracking, the main challenge is to maintain the correct identity of targets even under occlusions or when differences between the targets are small. The paper proposes a new approach to this problem by incorporating the context information. The context of a target in an image sequence has two components: the spatial context including the local background and nearby targets and the temporal context including all appearances of the targets that have been seen previously. The paper considers both aspects. We propose a new model for multitarget tracking based on the classification of each target against its spatial context. The tracker searches a region similar to the target while avoiding nearby targets. The temporal context is included by integrating the entire history of target appearance based on probabilistic principal component analysis (PPCA). We have developed a new incremental scheme that can learn the full set of PPCA parameters accurately online. The experiments show robust tracking performance under the condition of severe clutter, occlusions, and pose changes. PMID- 17108384 TI - Tracking people by learning their appearance. AB - An open vision problem is to automatically track the articulations of people from a video sequence. This problem is difficult because one needs to determine both the number of people in each frame and estimate their configurations. But, finding people and localizing their limbs is hard because people can move fast and unpredictably, can appear in a variety of poses and clothes, and are often surrounded by limb-like clutter. We develop a completely automatic system that works in two stages; it first builds a model of appearance of each person in a video and then it tracks by detecting those models in each frame ("tracking by model-building and detection"). We develop two algorithms that build models; one bottom-up approach groups together candidate body parts found throughout a sequence. We also describe a top-down approach that automatically builds people models by detecting convenient key poses within a sequence. We finally show that building a discriminative model of appearance is quite helpful since it exploits structure in a background (without background-subtraction). We demonstrate the resulting tracker on hundreds of thousands of frames of unscripted indoor and outdoor activity, a feature-length film ("Run Lola Run"), and legacy sports footage (from the 2002 World Series and 1998 Winter Olympics). Experiments suggest that our system 1) can count distinct individuals, 2) can identify and track them, 3) can recover when it loses track, for example, if individuals are occluded or briefly leave the view, 4) can identify body configuration accurately, and 5) is not dependent on particular models of human motion. PMID- 17108385 TI - Correspondence-free determination of the affine fundamental matrix. AB - Fundamental matrix estimation is a central problem in computer vision and forms the basis of tasks such as stereo imaging and structure from motion. Existing algorithms typically analyze the relative geometries of matched feature points identified in both projected views. Automated feature matching is itself a challenging problem. Results typically have a large number of false matches. Traditional fundamental matrix estimation methods are very sensitive to matching errors, which led naturally to the application of robust statistical estimation techniques to the problem. In this work, an entirely novel approach is proposed to the fundamental matrix estimation problem. Instead of analyzing the geometry of matched feature points, the problem is recast in the frequency domain through the use of Integral Projection, showing how this is a reasonable model for orthographic cameras. The problem now reduces to one of identifying matching lines in the frequency domain which, most importantly, requires no feature matching or correspondence information. Experimental results on both real and synthetic data are presented that demonstrate the algorithm is a practical technique for fundamental matrix estimation. The behavior of the proposed algorithm is additionally characterized with respect to input noise, feature counts, and other parameters of interest. PMID- 17108386 TI - Surface dependent representations for illumination insensitive image comparison. AB - We consider the problem of matching images to tell whether they come from the same scene viewed under different lighting conditions. We show that the surface characteristics determine the type of image comparison method that should be used. Previous work has shown the effectiveness of comparing the image gradient direction for surfaces with material properties that change rapidly in one direction. We show analytically that two other widely used methods, normalized correlation of small windows and comparison of multiscale oriented filters, essentially compute the same thing. Then, we show that for surfaces whose properties change more slowly, comparison of the output of whitening filters is most effective. This suggests that a combination of these strategies should be employed to compare general objects. We discuss indications that Gabor jets use such a mixed strategy effectively, and we propose a new mixed strategy. We validate our results on synthetic and real images. PMID- 17108387 TI - A statistics-based approach to binary image registration with uncertainty analysis. AB - A new technique is described for the registration of edge-detected images. While an extensive literature exists on the problem of image registration, few of the current approaches include a well-defined measure of the statistical confidence associated with the solution. Such a measure is essential for many autonomous applications, where registration solutions that are dubious (involving poorly focused images or terrain that is obscured by clouds) must be distinguished from those that are reliable (based on clear images of highly structured scenes). The technique developed herein utilizes straightforward edge pixel matching to determine the "best" among a class of candidate translations. A well-established statistical procedure, the McNemar test, is then applied to identify which other candidate solutions are not significantly worse than the best solution. This allows for the construction of confidence regions in the space of the registration parameters. The approach is validated through a simulation study and examples are provided of its application in numerous challenging scenarios. While the algorithm is limited to solving for two-dimensional translations, its use in validating solutions to higher-order (rigid body, affine) transformation problems is demonstrated. PMID- 17108388 TI - Topological equivalence between a 3D object and the reconstruction of its digital image. AB - Digitization is not as easy as it looks. If one digitizes a 3D object even with a dense sampling grid, the reconstructed digital object may have topological distortions and, in general, there exists no upper bound for the Hausdorff distance. This explains why so far no algorithm has been known which guarantees topology preservation. However, as we will show, it is possible to repair the obtained digital image in a locally bounded way so that it is homeomorphic and close to the 3D object. The resulting digital object is always well-composed, which has nice implications for a lot of image analysis problems. Moreover, we will show that the surface of the original object is homeomorphic to the result of the marching cubes algorithm. This is really surprising since it means that the well-known topological problems of the marching cubes reconstruction simply do not occur for digital images of r-regular objects. Based on the trilinear interpolation, we also construct a smooth isosurface from the digital image that has the same topology as the original surface. Finally, we give a surprisingly simple topology preserving reconstruction method by using overlapping balls instead of cubical voxels. This is the first approach of digitizing 3D objects which guarantees topology preservation and gives an upper bound for the geometric distortion. Since the output can be chosen as a pure voxel presentation, a union of balls, a reconstruction by trilinear interpolation, a smooth isosurface, or the piecewise linear marching cubes surface, the results are directly applicable to a huge class of image analysis algorithms. Moreover, we show how one can efficiently estimate the volume and the surface area of 3D objects by looking at their digitizations. Measuring volume and surface area of digital objects are important problems in 3D image analysis. Good estimators should be multigrid convergent, i.e., the error goes to zero with increasing sampling density. We will show that every presented reconstruction method can be used for volume estimation and we will give a solution for the much more difficult problem of multigrid-convergent surface area estimation. Our solution is based on simple counting of voxels and we are the first to be able to give absolute bounds for the surface area. PMID- 17108389 TI - Accurate and scalable surface representation and reconstruction from images. AB - We introduce a new surface representation method, called patchwork, to extend three-dimensional surface reconstruction capabilities from multiple images. A patchwork is the combination of several patches that are built one by one. This design potentially allows for the reconstruction of an object with arbitrarily large dimensions while preserving a fine level of detail. We formally demonstrate that this strategy leads to a spatial complexity independent of the dimensions of the reconstructed object and to a time complexity that is linear with respect to the object area. The former property ensures that we never run out of storage and the latter means that reconstructing an object can be done in a reasonable amount of time. In addition, we show that the patchwork representation handles equivalently open and closed surfaces, whereas most of the existing approaches are limited to a specific scenario, an open or closed surface, but not both. The patchwork concept is orthogonal to the method chosen for surface optimization. Most of the existing optimization techniques can be cast into this framework. To illustrate the possibilities offered by this approach, we propose two applications that demonstrate how our method dramatically extends a recent accurate graph technique based on minimal cuts. We first revisit the popular carving techniques. This results in a well-posed reconstruction problem that still enjoys the tractability of voxel space. We also show how we can advantageously combine several image-driven criteria to achieve a finely detailed geometry by surface propagation. These two examples demonstrate the versatility and flexibility of patchwork reconstruction. They underscore other properties inherited from patchwork representation: Although some min-cut methods have difficulty in handling complex shapes (e.g., with complex topologies), they can naturally manipulate any geometry through the patchwork representation while preserving their intrinsic qualities. The above properties of patchwork representation and reconstruction are demonstrated with real image sequences. PMID- 17108390 TI - The number of N-point digital discs. AB - A digital disc is the set of all integer points inside some given disc. Let {?cal D}_{N} be the number of different digital discs consisting of N points (different up to translation). The upper bound D(N) = O(N(2)) was shown recently; no corresponding lower bound is known. In this paper, we refine the upper bound to D(N) = O(N), which seems to be the true order of magnitude, and we show that the average [formula: see text] has upper and lower bounds which are of polynomial growth in N. PMID- 17108391 TI - Feature subset selection and ranking for data dimensionality reduction. AB - A new unsupervised forward orthogonal search (FOS) algorithm is introduced for feature selection and ranking. In the new algorithm, features are selected in a stepwise way, one at a time, by estimating the capability of each specified candidate feature subset to represent the overall features in the measurement space. A squared correlation function is employed as the criterion to measure the dependency between features and this makes the new algorithm easy to implement. The forward orthogonalization strategy, which combines good effectiveness with high efficiency, enables the new algorithm to produce efficient feature subsets with a clear physical interpretation. PMID- 17108392 TI - Dominant sets and pairwise clustering. AB - We develop a new graph-theoretic approach for pairwise data clustering which is motivated by the analogies between the intuitive concept of a cluster and that of a dominant set of vertices, a notion introduced here which generalizes that of a maximal complete subgraph to edge-weighted graphs. We establish a correspondence between dominant sets and the extrema of a quadratic form over the standard simplex, thereby allowing the use of straightforward and easily implementable continuous optimization techniques from evolutionary game theory. Numerical examples on various point-set and image segmentation problems confirm the potential of the proposed approach. PMID- 17108393 TI - A comparison of decision tree ensemble creation techniques. AB - We experimentally evaluate bagging and seven other randomization-based approaches to creating an ensemble of decision tree classifiers. Statistical tests were performed on experimental results from 57 publicly available data sets. When cross-validation comparisons were tested for statistical significance, the best method was statistically more accurate than bagging on only eight of the 57 data sets. Alternatively, examining the average ranks of the algorithms across the group of data sets, we find that boosting, random forests, and randomized trees are statistically significantly better than bagging. Because our results suggest that using an appropriate ensemble size is important, we introduce an algorithm that decides when a sufficient number of classifiers has been created for an ensemble. Our algorithm uses the out-of-bag error estimate, and is shown to result in an accurate ensemble for those methods that incorporate bagging into the construction of the ensemble. PMID- 17108394 TI - Variational surface interpolation from sparse point and normal data. AB - Many visual cues for surface reconstruction from known views are sparse in nature, e.g., specularities, surface silhouettes, and salient features in an otherwise textureless region. Often, these cues are the only information available to an observer. To allow these constraints to be used either in conjunction with dense constraints such as pixel-wise similarity, or alone, we formulate such constraints in a variational framework. We propose a sparse variational constraint in the level set framework, enforcing a surface to pass through a specific point, and a sparse variational constraint on the surface normal along the observed viewing direction, as is the nature of, e.g., specularities. These constraints are capable of reconstructing surfaces from extremely sparse data. The approach has been applied and validated on the shape from specularities problem. PMID- 17108395 TI - The role of melatonin receptor 1B gene (MTNR1B) in adolescent idiopathic scoliosis--a genetic association study. AB - Many studies have demonstrated the role of melatonin in the etiology of AIS. Previous studies have shown that there is no evidence of mutations in the melatonin receptor 1A gene in AIS patients. In this study, we have examined the role of melatonin receptor 1B in predisposition for AIS. Using haplotype block tagging technique, a set of tagging SNPs were defined for MTNR1B from the Han Chinese data of the International HapMap project. The association between the tagging of single nucleotide polymorphisms (tSNPs) in MTNR1B region and the occurrence of AIS was studied. METHOD: 473 AIS girls and 311 normal controls were recruited. The age range of the patients was between 10 and 18 years old. The maximum Cobb was recorded at latest follow-up in AIS patients. Three of five tSNPs were studied; they were all located within the coding region of the MTNR1B gene. RESULTS: There was no significant difference in the genotype or allelic frequencies (AF) of the 3 tSNPs between AIS and controls. In a case-only analysis, no difference in curve severity in AIS patients was found among patients with different genotypes (by one-way ANOVA). DISCUSSION: The 3 tSNPs showed no association with either the occurrence of AIS or the maximum Cobb angle within AIS girls. Further analysis of the remaining tSNPs within the regulatory region of the MTNR1B gene and other related genes in the melatonin signaling pathway may provide further information on the role of the melatonin in AIS girls. PMID- 17108396 TI - Correlation between cytogenetic abnormalities in cells and metabolic shifts in children with spinal deformities. AB - We studied the relationship between cytogenetic abnormalities in buccal epithelial cells and metabolic shifts in children with scoliosis and kyphosis. The incidence of nucleus abnormalities and the corresponding metabolic shifts were found to depend on the presence of spinal deformities and ecological factors. The problem of formation of risk groups for spinal deformities in ecologically unfavorable regions is discussed. PMID- 17108397 TI - Aggrecan gene expression disorder as aetiologic factor of idiopathic scoliosis. AB - A pathogenetic mechanism of the idiopathic scoliosis (IS) has been established on the basis of in-depth morphological and biochemical investigations of structural components of the spine in patients with IS (surgical material). We have shown that IS develops on the basis of disturbance of proteoglycans (PG) synthesis and formation in vertebral growth plates. The found keratan sulphate-related fraction is likely a marker of genetic changes in PGs in IS. Long-term our studies demonstrated a major-gene effect in IS. The study has shown that aggrecan gene expression is significantly decreased in cultivated chondroblasts from patients with IS. The presence of keratan sulphate-related fraction and keratan sulphate increase are associated with lumnican increase. PMID- 17108398 TI - Genetic association study of insulin-like growth factor-I (IGF-I) gene with curve severity and osteopenia in adolescent idiopathic scoliosis. AB - IGF-I has a pivotal role in bone growth and could be one of the putative disease modifier genes in AIS. Two SNPs in IGF-I gene promoter region were studied for any association with occurrence of AIS and for their effect on the curve severity among AIS. METHODS: 506 AIS girls (Cobb>20 degrees) and 227 age-matched Chinese girls were recruited. The spine (L2-L4) and hip BMD of the subjects were measured by DXA. A subgroup of AIS patients (N=340) who were followed-up to skeletal maturity and the maximum Cobb's angle was recorded. Two SNPs were genotyped by PCR-RFLP (rs5742612 and rs2288377). The chi-square test and one-way ANOVA were used to test the association between genotypes and quantitative parameters, respectively. RESULTS: No association was between the genotypes and the occurrence of AIS and the BMD of the spine and hip. The allelic frequency of T allele was 0.69 in AIS and control. However, the Cobb's angle was higher in patients with the homozygous T allele (Mean Cobb's angle: 38.1 degrees in TT vs 35.9 degrees in TC vs 33.2 degrees in CC group; p=0.04). DISCUSSION: Interestingly, IGF-I polymorphism affects the curve severity of AIS though it was not associated with onset of AIS per se. It indicates that IGF-I may be a disease modifying gene. The importance of IGF-I in skeletal growth makes it a good candidate gene which would play a role in the documented association of rapid growth with curve progression in AIS. PMID- 17108399 TI - Changes in vertebral neural arch morphometry and functional tethering of spinal cord in adolescent idiopathic scoliosis--study with multi-planar reformat magnetic resonance imaging. AB - With the use of multiplanar reformat Magnetic Resonance imaging, AIS patients were found to have significantly reduced pedicle widths on concavity. Pattern of vertebral asymmetry was also exaggerated with smaller pedicle width, length and area on concavity. The cord appeared more roundish and was deviated to the concavity at apical vertebra in AIS. A tethering force might therefore be present on the cord along the transverse axis in AIS, accounted by the relatively fixed position of the exit nerve roots and deviation of the cord from the exit foraminae of the corresponding vertebra. PMID- 17108400 TI - "Syndrome of contractures" (according to Mau) with the abduction contracture of the right hip as causative factor for development of the so-called idiopathic scoliosis. AB - The article provides basic explanation of "syndrome of contractures" (Mau) at newborns and babies and it's conjunction with biomechanical etiology of the so called idiopathic scoliosis (Karski 1995-2006). The authors analyzed children with "syndrome of contractures" and noted its relevance to some clinical symptoms at children with scoliosis. Newborns and babies with clinical signs of "syndrome of contractures" require further spine examination already at age of 3-4 in order to detect "danger of oncoming scoliosis" and to introduce neo-prophylaxis. The research based on "syndrome of contractures" can explain predominance of female gender of patients with scoliosis, sides of curves, side of rib hump, progression and sensibility to new rehabilitation exercises. PMID- 17108401 TI - Scoliotic progression patterns in prognostic factors and future prediction of spinal deformity progression. AB - This study investigated how an adolescent idiopathic scoliosis progresses with time. 154 consecutive measurements from 26 consecutive AIS patients were analyzed. Each subject had at least four successive scans at six-month intervals. Progression patterns of Cobb angle and apex lateral deviation were extracted from 34 serial data sets of the most common AIS type RT-LL, in the format of four serial data sets, by using the fuzzy c-means clustering technique. Progression of spinal deformity was predicted with previous serial data of Cobb angle and apex lateral deviation by using a GCV extrapolating technique alone and in conjunction with progression patterns. Our results showed that scoliotic progression appears to follow progression patterns. Progression of spinal deformity has potential to be accurately predicted with previous serial spinal deformities by using GCV extrapolating technique with assistance of progression patterns. PMID- 17108402 TI - Persistent osteopenia in adolescent idiopathic scoliosis--longitudinal monitoring of bone mineral density until skeletal maturity. AB - The aim of this study was to monitor BMD changes occurring during periods of rapid growth and to investigate whether osteopenia was a persistent phenomenon in skeletally matured AIS girls. 196 AIS Chinese girls and 122 healthy controls, aged 11-16, were follow-up for 3.5 years. Bilateral femoral neck bone mineral density (BMD) and volumetric BMD (vBMD) of the distal tibia were obtained by dual energy X-ray absorptiometry (DXA) and peripheral quantitative computed tomography (pQCT). Osteopenia was defined if the age-adjusted BMD was below or equal to -1 standard deviation (SD). The average age at the final follow-up was 16.8 years old. The median initial Cobb angle for this group of patients was 26 degrees. The prevalence of osteopenia at the baseline measurement was 35.9%. Longitudinal BMD results demonstrated that 86.0% of osteopenic AIS girls had persistently low BMD at the time of skeletal maturity (age of 16). vBMD of distal tibia of AIS was significantly lower than that of the controls throughout the age of 13 to 17 during the period of rapid growth. In addition, there were also significant differences in vBMD among AIS (moderate and severe group) and the controls by one way ANOVA (p<0.05). The present study for the first time revealed that over 86% of osteopenic AIS patients had persistently low BMD, at both distal tibia and femoral neck regions, at the time of skeletal maturity. Early detection and treatment of AIS-related osteopenia might help in maximizing peak bone mass during peripubertal growth that thereby minimizing risks of developing osteoporotic fractures later in life. PMID- 17108403 TI - Prevalence of scoliosis in women with visual deficiency. AB - Whether visual impairment influences the prevalence of scoliosis in humans or not remains controversial. The purpose of this study was to assess the prevalence of scoliosis in blind women in a Mediterranean region. MATERIAL AND METHOD: A total of 26 blind women aged 40 years (median, range 20 - 67) were screened for scoliosis. The existence of a possible trunk hump was measured by the forward bending test using the Pruijis Scoliometer. Reading of an Angle of Trunk Inclination (ATI) greater or equal to 7 degrees was used as a cut-off criterion for radiological examination. Standing postero-anterior and lateral spinal radiographs were obtained. Menarche and circadian rhythm was recorded. RESULTS: 11 out of 26 women had a scoliosis with an average Cobb angle of 19 degrees (range 12 - 28). The average ATI was 8 degrees. Thoracolumbar was the most common type of curve identified (9 out of 11, 6 were to the right and 3 to the left). The median age of menarche was 13 years (range, 11 - 15). None of the blind women reported any difficulty sleeping and had a circadian rhythm related to a 24-hour day. DISCUSSION: The prevalence of scoliosis was 42.3%, while the prevalence in the general population in the same regime is 2.9%. Blind women had a later age of menarche (13 versus 12.58 yrs) compared with normal girls. The postural etiology of scoliosis in blind people has been reported. The possible role of light in association to melatonin production, age at menarche and high prevalence of scoliosis in blind women is presented and discussed. PMID- 17108404 TI - Patterns of extra-spinal left-right skeletal asymmetries in adolescent girls with lower spine scoliosis: relative lengthening of the ilium on the curve concavity & of right lower limb segments. AB - Extra-spinal skeletal length asymmetry have been reported for the upper limbs and periapical ribs of patients with thoracic adolescent idiopathic scoliosis. This paper reports (1) a third pattern with relative lengthening of the ilium on the concavity of lower spine scolioses, and (2) a fourth pattern of relative lengthening of the right total leg and right tibia unrelated statistically to the severity or side of lower spinal scolioses. The findings pose the question: are these anomalous extra-spinal left-right skeletal length asymmetries unconnected with the pathogenesis of AIS. Or, are they indicative of what may also be happening to some vertebral physes as an initiating pathogenic mechanism for the scoliosis? PMID- 17108405 TI - Left-right upper arm length asymmetry associated with apical vertebral rotation in subjects with thoracic scoliosis: anomaly of bilateral symmetry affecting vertebral, costal and upper arm physes? AB - Left-right skeletal length asymmetries in upper limbs related to curve side and severity have been detected with adolescent idiopathic scoliosis (AIS). This paper reports upper arm length asymmetry in thoracic scoliosis related significantly to apical vertebral rotation in school screening referrals. The reason(s) for the association of upper arm length asymmetry with apical vertebral rotation is unknown and three factors are considered: (1) neuromuscular mechanisms from primary or secondary causes, (2) relative concave neurocentral synchondrosis overgrowth, and (3) relative concave periapical rib length overgrowth, A putative anomaly of growth plates (physes) of ribs, neurocentral synchondroses and upper arms, would account for the findings. A solution to this dilemma may emerge from the results of surgery should concave periapical rib resections become evaluated further for right thoracic AIS in girls. PMID- 17108406 TI - Etiologic theories of idiopathic scoliosis: neurodevelopmental concept of maturational delay of the CNS body schema ("body-in-the-brain"). AB - Several workers consider that the etiology of adolescent idiopathic scoliosis (AIS) involves undetected neuromuscular dysfunction. During normal development the central nervous system (CNS) has to adapt to the rapidly growing skeleton of adolescence, and in AIS to developing spinal asymmetry from whatever cause. Examination of evidence from (1) anomalous extra-spinal left-right skeletal length asymmetries, (2) growth velocity and curve progression, and (3) the CNS body schema, parietal lobe and temporoparietal junction, led us to propose a new etiologic concept namely of delay in maturation of the CNS body schema during adolescence. In particular, the development of an early AIS deformity at a time of rapid spinal growth the association of CNS maturational delay results in the CNS attempting to balance a lateral spinal deformity in a moving upright trunk that is larger than the information on personal space (self) already established in the brain by that time of development. It is postulated that the CNS maturational delay allows scoliosis curve progression to occur - unless the delay is temporary when curve progression would cease. The putative maturational delay in the CNS body schema may arise (1) from impaired sensory input: (2) primarily in the brain; and/or (3) from impaired motor output. Oxidative stress with lipid peroxidation in the nervous system may be involved in some patients. The concept brings together many findings relating AIS to the nervous and musculo-skeletal systems and suggests brain morphometric studies in subjects with progressive AIS. PMID- 17108407 TI - Intervertebral disc biomechanics in the pathogenesis of idiopathic scoliosis. AB - The aim of the present study is to investigate whether the deformation of the intervertebral disc contributes to the progression of idiopathic scoliotic curves. In the standing posteroanterior x-rays of 92 scoliotic curves the following readings were obtained: Cobb angle (CA), apical vertebral rotation (AVR), apical vertebral wedging (AVW) and the adjacent to the apical vertebra Upper (UIVDW) and Lower (LIVDW) InterVertebral Discs Wedging. The statistical analyses included inter - intraobserver reliability test, descriptives, monofactorial linear regression and Pearson correlation coefficient, with p<0.05 considered statistical significant (SS). The mean thoracic CA was 13.4 degrees, lumbar CA 13.8 degrees, thoracic AVR 5.3 degrees, lumbar AVR 4.7 degrees, thoracic AVW 1.4 degrees, lumbar AVW 1.5 degrees, thoracic UIVDW 1.6 degrees, thoracic LIVDW 1 degrees, lumbar UIVDW 1.3 degrees and lumbar LIVDW 2 degrees. Both thoracic and lumbar CA regressed SS with lumbar LIVDW, lumbar UIVDW, thoracic LIVDW and thoracic AVW. Lumbar LIVDW correlates SS with thoracic CA, lumbar CA and thoracic LIVDW. An inter and intra-observer error was below 1 degrees . The eccentric intervertebral disc in the scoliotic spine, through variation in its water concentration produces asymmetrically cyclical load during the 24-hour period and an asymmetrical growth of the vertebral body (Hueter Volkman's law). The statistical analysis revealed that AVW appears later when already CA increases, the IVDW is more important than AVW and the LIVDW, which is greater than UIVDW, is the most frequent correlated radiographic parameter. The deformation of the apical intervertebral disc seems to be an important contributory factor in the progression of a scoliotic curve. PMID- 17108408 TI - Geographic latitude and prevalence of adolescent idiopathic scoliosis. AB - Adolescent Idiopathic Scoliosis (AIS) prevalence has been reported to be different in various geographic latitudes and demonstrates higher values in northern countries. A study on epidemiological reports from the literature was conducted to record the prevalence of AIS among the general population of boys and girls, aged 10-16 years old, in different geographic latitudes, in order to test the hypothesis that the prevalence of AIS among boys and girls is different in various geographic latitudes and to examine if there is a possible association between them. Seventeen peer-reviewed published papers reporting AIS prevalence in the general population of boys and girls from most geographic areas of the northern hemisphere were retrieved from the literature. The geographic latitude of each centre where a particular study was originated was documented. The statistical analysis included a linear regression forward modeling procedure of the AIS prevalence by latitude, weighted by sample size. According to the modelling of the data, a significant positive association between prevalence of AIS and latitude was found for girls (p<0.001), following a rather curvilinear trend, but not a significant positive association was found for boys (p<0.111). A positive association between prevalence of AIS and geographic latitude is reported only for girls in the present study. Prevalence of AIS in boys is not associated significantly with geographic latitude. This differing significant association implicates the possible role of environmental factors in the pathogenesis of AIS that may act in a different way between boys and girls. PMID- 17108409 TI - Sagittal configuration of the spine in girls with idiopathic scoliosis: progressing rather than initiating factor. AB - Thoracic hypokyphosis with increasing axial rotational instability is claimed to be a primary factor for the initiation of Idiopathic Scoliosis (IS) according to some authors. The objective of this study was to compare the sagittal configuration of the spine in two groups of girls with and without scoliosis in order to determine whether thoracic hypokyphosis and/or lumbar hypolordosis are initiating factors for AIS or not. A group of 207 consecutive non-treated girls diagnosed with IS (12.7 y +/- 1.8) measured with the Formetric system were compared to a control group of 45 non-scoliotic girls of the same age (12.4 y +/- 2). The Cobb angle for the whole scoliosis sample was 26 degrees +/- 13.6 and the angle of axial rotation 12.4 degrees +/- 7.7 (Perdriolle). The patient group was divided into subgroups by their Cobb angle ie G1 (5 degrees -19 degrees, n=79), G2 (20 degrees -34 degrees, n=81), G3 (10 vertebral bodies, and with Cobb angle 45 degrees (p>0.05). The denervation of paraspinal muscles is present in some patients with scoliosis associated with syringomyelia, suggesting that scoliosis may be caused by a strength imbalance of paraspinal muscles in these patients. PMID- 17108414 TI - Quantitative analysis of types I and II collagen in the disc annulus in adolescent idiopathic scoliosis. AB - OBJECTIVE: To find the possible roles of collagen in the pathogenesis of AIS through studies of the distribution of the collagen I and II in the disc annulus fibrous in adolescent idiopathic scoliosis (AIS). METHODS: The disc annulus fibrous of apex curve were harvested during anterior surgery and divided into concave and convex samples. 25 AIS cases were covered in this study including 6 males and 19 females, with an average age of 14.6 years (range 12-18 years). 11 specimens were intervertebral disc from the thoracic region (from T8 to T11) and 14 sample were from lumbar discs (from L1 to L2). RT-PCR was employed to investigate the distribution and content of collagen I (220 bp) and collagen II (359 bp) in the intervertebral disc specimens. 1% agarose gel electrophoresis and the Gelwork image analysis system was used in the semi-quantitative analysis of the product. RESULTS: For the AIS group, type I collagen and type II collagen significantly increased on convex side compared with that of the concave side (p<0.01). The same trend was observed in both thoracic and lumber disc annulus fibrosis, but there was no statistical difference expect for the expression of type II collagen in convex side of disc annulus fibrous (p<0.05). CONCLUSION: The asymmetric expression of collagen I and II collagen showing that there was degeneration in the intervertebral disc of AIS. The abnormal collagen metabolism may be one of reasons in the development of AIS and probably an important factor in the progression of AIS. PMID- 17108415 TI - Asymmetric expression of melatonin receptor mRNA in bilateral paravertebral muscles in adolescent idiopathic scoliosis. AB - To investigate the change of melatonin receptor mRNA expression in bilateral paravertebral muscles in AIS, congenital scoliosis (CS) and controls in order to analyze its relationship to the pathogenesis of AIS. 20 cases with average age of 15.1 +/- 2.2 years and average Cobb angle of 56.2 degrees +/- 16.1 degrees were included in AIS group. 12 cases with average age of 11.6 +/- 3.2 years and average Cobb angle of 59.2 degrees +/- 33.3 degrees were included in congenital scoliosis (CS) group. 10 cases without scoliosis comprised a control group. The mRNA expression of melatonin receptor subtype MT1 and MT2 were detected by RT-PCR method. The MT2 mRNA expression on the concave side of paravertebral muscle was higher than that on the convex side in AIS and CS groups (p<0.05), but the MT1 mRNA expression showed no significant difference. In the AIS group, the ratio of MT2 mRNA expression on the concave side compared with the convex side in cases with a Cobb angle less than 50 degrees and cases with a Cobb angle greater than 50 degrees showed no significant difference. The melatonin receptor expression in bilateral paravertebral muscles in AIS is asymmetric, which may be a secondary change. PMID- 17108416 TI - Adolescent scar contracture scoliosis caused by back scalding during the infantile period. AB - To investigate the pathogenesis, clinical manifestation and treatment of the adolescent scar contracture scoliosis caused by back scalding during infancy. From August 1997 to May 2005, about 1300 patients with scoliosis received surgery in our department. Only four of them were diagnosed with adolescent scar contracture scoliosis. One patient was first treated with skin expansion, back scar excision, and skin flap transfer, followed with anterior correction with TSRH instrumentation. Two patients were first treated with back scar excision and anterior spinal release, then treated with posterior correction with TSRH instrumentation; thoracoplasty was performed after 50 days in halo-wheelchair traction. The other patient was treated with posterior correction with TSRH instrumentation. No management of scalding was performed on the fourth patient. Anterior release and posterior correction were performed at intervals of 3 weeks. The deformities of four patients were well corrected. The trunk balance was restored and the pelvis leveled. The skin incision wounds healed well. Minor loss of correction was recorded during the last follow-up. PMID- 17108417 TI - Classification of pediatric lumbosacral spondylolisthesis. AB - A surgical classification of pediatric lumbosacral spondylolisthesis has been proposed recently. In this classification involving 8 distinct types of spondylolisthesis, the patient is classified according to: 1) the slip grade (low vs. high-grade), 2) the degree of dysplasia (low- vs. high-dysplastic), and 3) the sagittal spinopelvic balance. The objective of this preliminary study is to assess the reliability of the classification. Two observers classified on two separate occasions 40 subjects with lumbosacral spondylolisthesis, based on standing postero-anterior and lateral radiographs of the spine and pelvis. No direct measurements on the radiographs were performed. All 8 types of spondylolisthesis were identified by the observers. Intra-observer agreement for the first and second observers was respectively 92.5% and 87.5%, while inter observer agreement was 75.0%. Thirty-nine of 40 subjects had agreement among both observers according to the slip grade. Within these 39 subjects, observers disagreed for 8 subjects with respect to the degree of dysplasia and for only one subject with respect to the spinopelvic balance. The proposed classification could be used to better evaluate and compare available surgical techniques, and to develop a treatment algorithm for spondylolisthesis. This new classification results in good intra- and inter-observer agreement. Further studies with observers not involved in the design of the classification are however needed in order to confirm the relevance of the classification. PMID- 17108418 TI - Hoffmann reflex in idiopathic scoliosis. AB - This study was carried out in order to determine the dependence of selected Hoffmann reflex parameters on type, progression and morphology of idiopathic scoliosis (IS). Data collected from 129 girls with IS (59 progressive and 70 non progressive cases) aged 7-16 years and 24 healthy subjects were analysed. H reflex index (IH) and H/M amplitude index (IH/M) were calculated. Progressive left lumbar scoliosis expressed a significant decrease of IH values and a distinct tendency to IH/M depletion compared to non-progressives and controls. Progressive right thoracic scoliosis expressed marked tendency to IH decrease compared to nonprogressive scoliosis. No significant differences in H-reflex parameters were observed between the convex and concave side of the curvature or between types of scoliosis. H-reflex analysis in idiopathic scoliosis supports the hypothesis of a primary neurological disorder in progressive IS. PMID- 17108419 TI - Indoor heated swimming pools: the vulnerability of some infants to develop spinal asymmetries years later. AB - Evidence reported in an earlier paper suggests that infants introduced to indoor heated swimming pools in the first year of life show an association with spinal asymmetries including progressive adolescent idiopathic scoliosis (AIS) and in normal subjects vertical spinous process asymmetry. Indoor heated swimming pools may contain a risk factor that predisposes some infants to develop such spinal asymmetries years later. What the risk factor(s) may be and its possible portal of entry into the infant's body are unknown and possibilities are examined. New teenage controls were obtained after mothers of AIS patients mentioned that they had taken their child to an infant swim class. In a further group of 18 normal teenagers introduced to an indoor heated swimming pool in the first year of life, 15 had vertical spinous process asymmetry. This prevalence of 83% of those at risk confirms our previous observation of vertical spinous process asymmetry in 61% of teenagers who were introduced to indoor heated swimming pools in the first year of life. Subject to confirmation of our observations consideration should be given to chemical risk factors, possible portals of entry, toxicology, environmental epigenomics and disease susceptibility to altered spinal development. If the risk factor is confirmed there may ultimately be a place for the prevention of AIS in some subjects. PMID- 17108420 TI - Effect of same-sided and cross-body load carriage on 3D back shape in young adults. AB - Regular carriage of heavy loads such as backpacks, satchels and mailbags results in a variety of acute medical problems and increased potential for back injury. There is a paucity of information about the specific changes in back posture that occur in response to asymmetrical loading. The purpose of this study was to examine the changes in back shape that occurred in response to asymmetrical load carriage, either on one shoulder (same-side) or across the body (cross-body), in healthy young adults. METHODS: A convenience sample of 21 physiotherapy students randomly performed three trials (unloaded, same-side loaded, cross-body loaded) in standing with a 15% body load. The Microscribe 3DX digitiser (Immersion Group Ltd) recorded the three dimensional coordinates of 15 Key anatomical landmarks on the back in the three conditions. RESULTS: A one-way ANOVA with repeated measures and post-hoc tests was implemented to highlight statistical differences in the data collected (p<0.05). Significant differences were found in the x, y and z coordinates of the anatomical landmarks in the upper back between unloaded and loaded conditions. Results demonstrated significantly less impact on spinal posture from cross-body loading as compared to same-sided loading. CONCLUSION: This study confirms that there are significant three-dimensional changes in back shape in response to asymmetrical loading. Further work is needed to evaluate the optimal carriage type and maximal body load that results in the least spinal impact and injury potential in young adults. PMID- 17108421 TI - Transverse plane of apical vertebra of structural thoracic curve: vertebra displacement versus vertebral deformation. AB - CT transversal scans of the trunk provided at the level of Th8 or Th9 (apical vertebra) of 23 patients with structural thoracic scoliosis were reviewed. The following parameters were studied: 1) alpha angle formed by the axis of vertebra and the axis of spinous process, 2) beta concave and beta convex angle between the spinous process and the left and right transverse process respectively, 3) gamma concave and gamma convex angle between the axis of vertebra and the left and right transverse process respectively, 4) rotation angle to the sagittal plane according to Aaro and Dahlborn, 5) Cobb angle. Values of measured parameters demonstrated a common pattern of intravertebral deformity: counter clockwise deviation of the spinous process (alpha angle 15,0 +/-8,5 degrees), beta concave (69,8 +/-8,5 degrees) significantly greater than beta convex (38,8 +/-8,5 degrees), gamma concave (54,3 +/-7,8 degrees) not different from gamma convex (56,0 +/-8,0 degrees). Strong linear positive correlation between alpha angle and Aaro-Dahlborn angle was observed (r=0,78, p<0,05). Changes in morphology of apical vertebra due to intravertebral bone remodelling followed the vertebral spatial displacement and there existed a linear correlation in between. The two processes develop in opposite directions. PMID- 17108422 TI - Correlation of an induced rotation model with the clinical categorisation of scoliotic deformity--a possible platform for prediction of scoliosis progression. AB - AIM: The primary aims were to develop a simplified three-dimensional model of the thoracolumbar spine and to predict the influence of axial rotation at different levels of the spine on the resultant scoliotic curve, using King's classification as a comparator. MATERIALS AND METHODS: A three-dimensional mathematical model of the simplified thoracolumbar spine (constant size vertebral body without posterior elements) was developed using anatomical data from the published literature. The influence of rotational displacement of the motion segments at various levels of the spine was studied by applying different axial rotations, using a three-dimensional homogeneous transformation matrix method. RESULTS: The result of the model show the correlation of the deformity in lower regions of the spine (lumbar) with the geometrical changes in upper regions of the spine (thoracic), associated with the continuous alteration in direction of the vertebral axis of rotation along the spine. The final curvature of the scoliotic spine is influenced by both the degree of axial rotation in each region and the spatial deformation of the spine (e.g. kyphotic shape and extent of lateral deformity). Qualitatively, the model is capable of producing different categories of the spinal deformity based on King's classification. CONCLUSION: A three dimensional analysis of spinal shape demonstrates the important relationship between induced vertebral rotation and the resulting deformity. The effect of rotational displacement on the overall configuration of thoracolumbar spine during juvenile growth was assessed and demonstrates close correlation with deformities of the lower regions of spine. PMID- 17108423 TI - A novel framework for the 3D analysis of spine deformation modes. AB - Three-dimensional classification of scoliosis is important. However, analyzing large databases of 3D spine models is a difficult and time consuming task. To facilitate this task a method that automatically extracts the most important deformation modes from a set of 3D spine models is proposed. The 3D spine models are first converted into vertebrae relative positions and orientations. Then, a variability model composed of the Frechet mean and of a generalized covariance is computed. A principal component analysis is applied to that variability model and the extracted components are converted into deformation modes. Those modes are visualized by animating a 3D spine model where the deformation strength varies (for a given mode). The proposed method was applied to a group of 307 scoliotic patients and meaningful deformation modes were successfully extracted. For example, patients' growth, double curves, simple thoracic curves and lumbar lordosis were extracted in the first four deformation modes. Moreover, the obtained deformation modes are not disconnected from conventional surgical classifications since a logistic regression confirmed that there is a statistically significant relation between King's classification and the first four principal deformation modes. The proposed method successfully extracted important deformation modes from a set of 3D spine models and can be used to refine arbitrary classes (King's or Lenke's classes, for instance), thus helping the design of new clinically relevant 3D classifications. PMID- 17108424 TI - Intervertebral disc adaptation to wedging deformation. AB - Although scoliosis includes wedge deformities of both vertebrae and discs, little is known about the causes of the discal changes, and whether they result from mechanical influences on growth and/or remodelling. METHODS AND MATERIALS: An external apparatus attached to transvertebral pins applied compression and 15 degrees of angulation to each of two adjacent young rat caudal intervertebral discs for 5 weeks (four animals), or for 10 weeks (four animals). Each week, micro-CT scanning documented the in vivo discal wedging. After euthanasia, tail segments (three vertebrae and the 2 angulated discs) were excised and their flexibility was measured over a range of lateral bending. The angle of maximum flexibility was recorded. Then discs were fixed in situ (with the external apparatus in place) and sectioned for polarized light microscopy. RESULTS: The disc-wedging deformity averaged 15 degrees initially, it averaged 20 degrees after 5 weeks, and then reduced to 10 degrees (in 10 week animals). The lateral bending flexibility showed a distinct maximum at an average of 1.1 degrees from the in vivo position in the 5-week animals, indicating structural remodeling of the discs almost to the deformed geometry. The 10-week animals had maximum flexibility at 1.4 degrees from the in vivo position (no significant difference between 5 and 10-week animals.) Collagen crimp angles [Cassidy et al., Conn Tiss Res 1989, 23:75-88] were not significantly different between convex and concave sides, again suggesting that remodeling had occurred. CONCLUSIONS: In a mechanically induced scoliosis deformity in skeletally immature rats, the intervertebral discs underwent remodeling within 5 weeks. This indicates that this animal model is suitable for studying adaptive wedging changes in human scoliosis. PMID- 17108425 TI - A 3-D biomechanical skeleton model for posture and movement analysis. AB - A project to merge into a full 3D reliable and detailed human skeleton representation various segmental biomechanical models presented in literature has been undertaken by our group. The obtained 3D skeleton model is fully parametric and can so be fitted to each subject anthropometric characteristics. A non ionizing approach based on 3D opto-electronic measurements of body landmarks labelled by passive markers has been chosen to build the 3D parametric biomechanical skeleton model. To this aim various protocols involving different body labelling (and so different related anthropometric data) have been established for different analyses. To analyse human posture and spinal related pathologies, a 27 markers protocol has been set for static analysis, while 49 markers protocol has been set for gait and movement analysis. A special focus has been devoted to identify and model the spine with a correct degree of accuracy and reliability. To this aim complex signal processing and optimisation procedures have been tested. The model is able to fully integrate information deriving from other measurements devices as force platform data, surface EMG, foot pressure maps. The presented model is the first proposed in literature, to authors knowledge, able to process such multifactorial information to perform a full kinematic and kinetic analysis with particular focus on the spine. Several hundreds of patients have been already analysed and followed up with this methodology that proved to be useful for various posture and spine related pathologies (in particular spine deformities, low-back pain etc.). PMID- 17108426 TI - Comparison of sacropelvic morphology between normal adolescents and subjects with adolescent idiopathic scoliosis. AB - Previous studies suggest that the pelvic morphology may be abnormal in adolescent idiopathic scoliosis. This study compares the sacropelvic morphology between normal adolescents and subjects with adolescent idiopathic scoliosis (AIS) in both sagittal and coronal planes. The sacropelvic morphology was assessed from the postero-anterior and lateral standing radiographs of 27 normal adolescents and 29 subjects with AIS presenting a major Cobb angle greater than 30 degrees . Sacropelvic morphology was characterized by 19 parameters in the sagittal plane and 26 parameters in the coronal plane. There was no difference in sacropelvic morphology between the two groups in the sagittal plane. In the coronal plane, significant differences were found for right pelvic length, right iliac height, left and right pubic length, left obturator foramen width, bicristal distance, bituberal distance, biacetabular distance, pubic symphysis width, pelvic inlet, and subpubic angle. There was no significant pelvic asymmetry in AIS subjects. This is the first study that specifically evaluates the sagittal and coronal sacropelvic morphology in AIS. The results suggest that the coronal sacropelvic morphology is distorted in AIS. A longitudinal study is required in order to evaluate the influence of sacropelvic morphology in the progression of AIS. PMID- 17108427 TI - Can posture analysis point towards curve progression in scoliotic subjects? AB - Previous research employing biomechanical measurement has demonstrated asymmetries in kinematics and kinetics. Similar asymmetries have been reported from anthropometric studies. These findings suggest that asymmetry may play an important aetiological role in adolescent idiopathic scoliosis (AIS). The present study is a part of a wider comprehensive investigation aimed at identifying asymmetries in lower limb kinematics and pelvic and back movements during level walking in a sample of scoliotic subjects. Such asymmetries may be related to the spinal deformity. While previous studies indicate that force platform measurements provide a good estimation of the static balance of individuals, there remains a paucity of information on dynamic balance during walking. There is published evidence on the use of Centre of Pressure (CoP) and net joint moments in gait assessment. Although these investigations have assessed Centre of Mass (CoM)-CoP distance relationships in clinical conditions, there is a paucity of data relating to the moments about CoM. An objective of the present study was to assess and establish the asymmetry in the CoP pattern and moments about CoM during level walking and its relationship to spinal deformity. Results indicate differences across the subjects depending on the laterality of the major curve. Furthermore, the results indicate that the variables identified in this study could be applied to initial screening and surgical evaluation of scoliosis and other spinal deformities. Further studies are being undertaken to validate these findings. PMID- 17108428 TI - Validity and reliability of active shape models for the estimation of Cobb angles in adolescent idiopathic scoliosis. AB - Choosing the most suitable treatment for scoliosis relies heavily on accurate and reproducible Cobb angle measurement from successive radiographs. The objective is to reduce variability of Cobb angle measurement by reducing user intervention and bias. Custom software to automate Cobb angle measurement from posteroanterior radiographs was developed using active shape models. Validity and reliability of the automated system against a manual and semi-automated measurement method was conducted by two examiners each performing measurements on 3 occasions from a test set (N=22). A training set (N=47) of radiographs representative of curves seen in a scoliosis clinic was used to train the software to recognize vertebrae from T4 to L4. Images with a maximum Cobb angle between 20 degrees and 50 degrees, excluding surgical cases, were selected for training and test sets. Automated Cobb angles were calculated using best-fit slopes of the detected vertebrae endplates. Intra-class correlation coefficient (ICC) and standard error of measurement (SEM) showed high intra-examiner (ICC > 0.90, SEM 2-3 degrees) and inter-examiner (ICC > 0.82, SEM 2-4 degrees), but poor inter-method reliability (ICC=0.30, SEM 8-9 degrees). The automated method underestimated large curves. The reliability improved (ICC = 0.70, SEM 4-5 degrees) with exclusion of the 4 largest curves (>40 degrees) in the test set. The automated method was reliable for moderate sized curves, but did not properly detect vertebrae in larger curves. Optimization of constraints on scaling, rotation, translation, and iteration may improve reliability with larger curves. PMID- 17108429 TI - Surface alignment to unmask scoliotic deformity in surface topography. AB - Comparisons of back surface topography to monitor scoliosis are hampered by shape change due to other causes, primarily stance variations. The aim is to extract changes due only to the scoliosis from the observed changes. Surface (back) topography data were derived from 2 patients measured repeatedly (10x) at a single visit and from 45 patients measured at routine clinical visits. The measured back shapes were aligned to obtain the position of closest fit using a mathematical technique. The shape similarities were then quantified with a closeness-of-fit index. The similarity indices from the alignments were used to estimate noise factors due to postural variation, relative to typical patient change. Surface alignment has also been trialled using models with predictable topographic changes to determine whether the noise sources can be modelled. The values of the root-mean-square of the residual surface differences at all measured points after mathematical back shape alignment were used as a surface similarity index. The magnitude of this index for the repeatedly measured back shapes indicates the back shape variation due to stance, and has been compared with the variation found in normal patients. The similarity index has also been examined for cases both with and without modelled changes to estimate the effectiveness of stance modelling. The noise due to stance change was indicated by a surface similarity index which averaged 2.0 mm; the index for backs measured at routine visits averaged 3.5 mm. It was concluded that noise due to stance is significant and deserves to be recognised, in the comparison of back surfaces in three-dimensions. There is evidence that changes due to noise can be modelled. PMID- 17108430 TI - Automatic matching of spine images to assess changes in scoliosis. AB - This paper presents an image matching approach that can be used to measure changes in scoliotic curves. The proposed approach uses a novel fuzzy logic controller to estimate all open parameters. Using fluoroscopy images of a spine phantom, it was found that, with minimal user interaction, the matching of spine images could be achieved with high accuracy (the average errors were around 0.03 mm) and high computational efficiency (requiring less than 1 minute for matching each vertebra). PMID- 17108431 TI - A comparison of the clinical effectiveness of spinal orthoses manufactured using the conventional manual method and CAD/CAM method in the management of AIS. AB - Spinal orthoses are commonly prescribed to patients with moderate AIS for prevention of further deterioration. In a conventional manufacturing method, plaster bandages are used to get patient's body contour and plaster cast is rectified manually. With the introduction of CAD/CAM system, a series of automated processes from body scanning to digital rectification and milling of positive model can be performed in a fast and accurate fashion. This project is to study the impact of CAD/CAM method as compared with the conventional method. In assessing the 147 recruited subjects fitted with spinal orthoses (43 subjects using conventional method and 104 subjects using CAD/CAM method), significant decreases (p<0.05) were found in the Cobb angles when comparing the pre intervention data with that of the first year of intervention. Regarding the learning curve, Orthotists are getting more competent with the CAD/CAM technique in four years time. The mean productivity of the CAD/CAM method is 2.75 times higher than that of the conventional method. The CAD/CAM method could achieve similar clinical outcomes and with its high efficiency, could be considered as substitute for conventional methods in fabricating spinal orthoses for patients with AIS. PMID- 17108432 TI - The accumulation of bone mineral content and density in idiopathic scoliotic adolescents treated with bracing. AB - e aimed to investigate the accumulation of bone mineral content (BMC) and bone mineral density (BMD) during bracing treatment in patients with adolescent idiopathic scoliosis (AIS). Using a dual energy x-ray absorptiometer, initial and follow-up BMC and BMD measurements were taken in the lumbar spine (L2-L4) and the dominant (left) femoral neck of a total of 40 AIS patients who were treated with braces. Using a paired 't' test, both BMC and BMD of both sites, at the follow-up of about 6 months and 12 months, were significantly higher than those right before bracing treatment (p<0.05). BMD measured at all sites increased at a rate similar to reported normal values in patients who had normal BMD as well as those who had lower initial BMD. No significant correlation was found between increase in BMC and BMD and daily brace wear (p>0.05). In conclusion, both BMC and BMD levels grew during brace treatment in AIS at a speed similar to reported normal values, suggesting that bracing dose not appear to adversely affect the accumulation of bone mass in AIS. PMID- 17108433 TI - Prediction of brace treatment outcomes by monitoring brace usage. AB - Brace treatment is the most commonly used non-surgical treatment method for adolescent idiopathic scoliosis (AIS). This study determined whether curve progression can be predicted by how often and how well children with AIS wear their braces. Twenty subjects (3M, 17F) who were diagnosed with AIS and had worn their braces from six months up to 1 year participated into this study. All subjects were prescribed Boston style braces and have now completed their brace treatment. On average, the brace was used 57% of the prescribed time. Peterson's risk of progression (Risser sign, age, apex of curve and imbalance of curve) predicted only 3-8% of the curve progression of brace subjects. Knowing how brace subjects used their braces in terms of brace tightness increases the prediction rate to 12-21%; and wear time further increase it to 25-36%. Adding the multiple of brace tightness and wear time improves curve progression prediction to 41-54%. To be most effective, the brace should be worn as prescribed in both tightness and time manners. PMID- 17108434 TI - The Sforzesco brace and SPoRT concept (Symmetric, Patient-oriented, Rigid, Three dimensional) versus the Lyon brace and 3-point systems for bracing idiopathic scoliosis. AB - We have developed a new rigid brace, the Sforzesco brace, according to a new concept SPoRT (Symmetric, Patient-oriented, Rigid, Three-dimensional, active). The aim of this study is to verify the first results of the SPoRT concept compared to a classical 3 point system. DESIGN: A pair-controlled retrospective study. POPULATION: 15 Patients (22 females), 14.0+/-1.7 years, 39.8+/-9.8 degrees Cobb ( degrees C). We examined for differences between the two groups, at the start with SPoRT worst than LY. All degrees C and degrees B parameters (maximal, average and single localizations) decreased significantly in both groups, apart from thoracic degrees C in LY. SPoRT had better results than LY (P<0.05) for maximal degrees C and average degrees C. Moreover, degrees C clinical results were better (P<0.05) in SPoRT than LY. SPoRT had better results than LY in aesthetic (P<0.05) of the shoulders and waists. Brace is effective in the short term also in high degree curves, and SPoRT obtained statistically significant 80% better degrees C results than LY in the worst scoliosis curve, and 40% in the average of all curves, as well as better aesthetic. PMID- 17108435 TI - The ScoliOlogiC "Cheneau light" brace--does the reduction of material affect the desired correction? AB - The "Cheneau light" brace, developed according to the Cheneau principles, promises a lesser impairment of quality of life in the brace. However material reduction should not result in reduced effectiveness. Therefore the primary correction effect in the "Cheneau light" brace has been evaluated and compared with that of other traditional braces. MATERIAL AND METHOD: The correction effects of the first 99 patients (main diagnosis AIS or EOS; 7 boys, 92 girls), treated according to the principles of the "Cheneau light" brace were evaluated after an average treatment time of 6 weeks by a full-body X-ray made in the standing position while wearing the brace and compared with the last X-ray before bracing. The average curvature angle of the whole group was 41 degrees , the average age was 13 years. RESULTS: The Cobb angle in the whole sample has been reduced by an average of 14,7 degrees, which corresponds to a correction effect of 42%. In patients from this sample who had their first brace (n=53; Cobb angle 36,6 degrees) the in-brace correction was at 49,8%. The correction effect correlated slightly negative with age (r = -0,18; p = 0,034) and correlated negatively with the Cobb angle measured before treatment (r = -0,49; p = 0,0001). CONCLUSIONS: The use of the "Cheneau light" brace leads to correction effects above average when compared to correction effects of other braces described in literature. The reduction of material seems to affect the desired correction in a positive way. PMID- 17108436 TI - Biomechanical modeling of brace design. AB - OBJECTIVE: To study the biomechanical effectiveness of brace design parameters in right thoracic idiopathic scoliosis. METHODS: A finite element model (FEM) of the spine, rib cage, pelvis and abdomen was adapted to the geometry of 8 patients with right-thoracic idiopathic scoliosis using a multi-view radiographic reconstruction technique. A detailed parametric FEM of a thoraco-lumbo-sacral orthosis and a Box, Hunter & Hunter experimental design method were used to analyze the contribution of brace design parameters (brace size, number of straps, strap tension, position of the thoracic pad, lordosis reduction design) and of patient's spine stiffness. RESULTS: The mean Cobb angle correction of the thoracic curve was 5.1 degrees (0 degrees to 16 degrees). The most influential parameters were, in descending order, the strap tension, lordosis reduction design and spine stiffness. Their effects are independent and remain weak (-3 degrees when strap tension increases from 20 N to 60 N). Changing the position of the thoracic pad (slightly above or below the apex) doesn't have a significant effect. No significant correction of the axial rotation and rib hump was obtained. DISCUSSION & CONCLUSION: Frontal curve correction varied significantly, which justifies the need for an adequate adjustment of the brace. A more efficient design for the correction of transverse deformities remains to be found. The "active" correction component by the muscles was not included, but one can anticipate that its action would be concurrent to the passive brace mechanisms, enabling supplementary correction. A new tool simulating brace treatment has been developed, which allows rational design of braces. PMID- 17108437 TI - Spinal hemiepiphysiodesis correlates with physeal histomorphometric gradients. AB - Compression of a growth plate is known to correlate with changes to growth plate structure. The purpose of this study was to determine if the height of the physeal hypertrophic zones and chondrocyte size were directly related to the distance from a spine implant. The implant was designed with the goal of slowing spine growth asymmetrically. For stapled vertebrae, hypertrophic zone height, cell height and cell width were decreased across the coronal plane of the growth plate, with the lowest values under the staple, 8 weeks postoperatively. PMID- 17108438 TI - Radiological results of Dobosiewicz method of three-dimensional treatment of progressive idiopathic scoliosis. AB - The aim of the study was the radiological evaluation of the three-dimensional asymmetric treatment of scoliosis in strict symmetric initial positions. MATERIAL AND METHOD: 152 Patients (137 F and 15 M) with idiopathic progressive scoliosis (85 single scoliosis cases and 67 double-major) with proven curve progression were retrospectively analysed on the basis of radiological evaluation in the period of years 1999-2004. Mean age of assessed group was 14.22 years (range 7/21, SD=2.57), mean time of observation 31.80 months (range 18/63, SD=10.75). All children were treated with Dobosiewicz method of three-dimensional asymmetric correction in strict symmetric initial positions and with rigid Cheneau brace when indicated. RESULTS: Mean initial value of Cobb angle was 25.92 degrees (range 9/62, SD=10.59), mean initial rotation of apical vertebra was 9.55 degrees (range 0/30, SD=7.66). The outcome values were 31.04 degrees (range 5/76, SD=13.96) and 12.97 degrees (range 0/35, SD=8.41), respectively. Mean progression of Cobb angle during entire period of observation was 5.12 degrees (range -21 / +38, SD=9.62), mean progression of apical vertebra rotation was 3.42 degrees (range -21 / +25, SD=6.03). Further multi-factor analysis is discussed in full text. Results are also presented in the form of radiograms. CONCLUSION: The radiological results demonstrated prevalent stabilisation of scoliotic curves in children treated with Dobosiewicz method between October 1999 and December 2004. PMID- 17108439 TI - Trunk rotational strength training for the management of adolescent idiopathic scoliosis (AIS). AB - Quantified trunk rotational strength training has shown promise as a non operative management option for individuals with AIS. The purposes of our study are to test whether a quantified trunk rotational strength training protocol can increase trunk strength and stabilize or decrease curve size. Seven adolescents with AIS (5 female 2 male; mean 14 yrs +/- 2.6 yrs; mean Cobb 28 degrees +/- 6 degrees range 20 degrees -37 degrees) underwent four months of supervised trunk rotational strength training, and repeat strength test. Trunk strength in both directions increased significantly after training (p<0.05). Average Cobb angle decreased to 23 degrees +/- 11 degrees (range 6 degrees -35 degrees). Four individuals showed reduction (>5 degrees) in their original curve, and 3 remained the same (+/-5 degrees). The strength training protocol significantly increased isometric rotational strength and scoliosis was stabilized short term. PMID- 17108440 TI - Multiple vertebral wedge osteotomy for adolescent idiopathic scoliosis. AB - 19 idiopathic and 1 syringomyelia scoliosis patients (17 females and 3 males) underwent fusionless, multiple vertebral wedge osteotomy and with follow-up for an average of 8.9 years. The average age at surgery was 16.4 years. The average curve magnitude measured by Cobb was 64.0 degrees before surgery, 38.8 degrees after surgery, 43.7 degrees one year after surgery, 46.9 degrees two years after surgery and 48.2 degrees at the latest follow-up. The difference between the Cobb angle two years after surgery and latest follow-up was not significant. There were no major complications such as neurological problems. PMID- 17108441 TI - Experimental usage of hydroxyapatite preadsorption with fibronectin to increase permanent stability and longevity of spinal implants. AB - Hydroxyapatite has been used in orthopaedic and particularly in spinal surgery by precoating implants to indirectly increase osteoblasts' adhesion and subsequently their stability and longevity. Fibronectin preadsorption synergistically with appropriately constructed hydroxyapatite's surface texture to enhance osteoblasts' adhesion has not been, to the authors' knowledge, previously investigated. In osteoporotic spines, methods to increase implant stability (pedicle screws and cages) are of major value. OBJECTIVE: This experimental study investigated the contribution of fibronectin preadsorption to enhance osteoblasts' adhesion and strength on hydroxyapatite. METHODS: Hydroxyapatite substrata with two different surface roughnesses (rough HA180 and the smooth HA1200) were produced and human osteoblasts were seeded on them after culture. Prior to osteoblasts seeding, the hydroxyapatite substrata were immersed in fibronectin solution. Osteoblast attachment on each of the two hydroxyapatite substrata was evaluated by recording the number of cells, while the osteoblast's adhesion strength was determined by measuring the shear stress required to detach the cells from the hydroxyapatite substrates. RESULTS: Fibronectin preadsorption increased the number of attached osteoblasts on smooth and rough hydroxyapatite substratum at 40% and 62% respectively, while it increased osteoblast attachment strength on the smooth and rough substratum at 165% and 73% respectively. CONCLUSIONS: Fibronectin preadsorption and smooth hydroxyapatite surface texture synergistically increased the adhesion's strength of human osteoblasts "in vitro", while preadsorption and rough hydroxyapatite surface increased the number of attached osteoblasts. Further studies in primates and human beings should be carried out to disclose the clinical relevance of the above mentioned observations in spine surgery. PMID- 17108442 TI - Relation between the sagittal pelvic and lumbar spine geometries following surgical correction of adolescent idiopathic scoliosis: a preliminary study. AB - The influence of surgery on the relationship between the lumbar spine and the pelvis in adolescent idiopathic scoliosis (AIS) remains mostly unknown. The sagittal spinopelvic balance of 40 patients with AIS undergoing posterior spinal instrumentation and fusion was studied. After surgery, the total lumbar lordosis (LL) and the LL below fusion remained correlated to the sagittal pelvic geometry. LL within fusion, which is set by the instrumentation, was also related to the LL below fusion. This preliminary study shows that pelvic morphology probably has an important impact on the postoperative sagittal lumbar alignment. Evaluation of the sagittal pelvic geometry could therefore be useful in the preoperative planning of surgery for AIS. Further studies with more patients are still needed in order to confirm this hypothesis and to evaluate if the distal fusion level influences the spino-pelvic balance. PMID- 17108443 TI - Timing of osteotomy for thoracolumabar or lumbar kyphosis secondary to ankylosing spondylitis. AB - OBJECTIVE: Ankylosing spondylitis may lead to a rigid thoracolumbar kyphotic deformity. Several authors have reported the results of patients treated by a lumbar osteotomy, but there is no consensus on the level of the osteotomy and on timing of osteotomy. The purpose of this study is to explore timing of osteotomy for thoracolumabar or lumbar kyphosis secondary to ankylosing spondylitis by analyzing the natural history of 78 AS patients. METHOD: To analyze the factors related to influence the timing of osteotomy for thoracolumabar or lumbar kyphosis secondary to ankylosing spondylitis. Patient Sample There were 78 patients including 69 male and 9 female; the mean age at the time of surgery was 38 years (range, 22-56 years). The characteristic of natural history of these 78 patients is lumbar pain stage, slowly progressive kyphosis stage, accerlated progressive kyphosis stage, stabilized kyphosis stage. The average preoperative deformity was 61 degrees (range, 40 degrees to 87 degrees). Twenty-eight patients underwent V shape osteotomy, and 50 patients underwent transpedicular osteotomy. RESULT: The average age of occurrence of lumbar pain symptom is 21 years,the average duration of slowly progressive kyphosis stage is 3 years (range, 1 to 8 year), and the average duration of accerlated progressive kyphosis stage is 4 (range, 2 to 10 year). The patients were followed up for a period of 4-60 months (mean 49 months). The total correction postoperatively was 40 degrees +/-11 degrees, the average loss of correction was 5 degrees at the final follow-up. Excellent results were obtained in 74 patients (95%), and good results were obtained in 4 patients (5%), there were no fair or poor results. CONCLUSION: Lumbar pain arrests for more than 6 months (exclusive of mechanical pain of lumbar), normal blood sedimentation rate continues for two times, and reactive protein is negative. These factors must be considered in timing of osteotomy for thoracolumabar or lumbar kyphosis secondary to ankylosing spondylitis. PMID- 17108444 TI - Biomechanical assessment of variable instrumentation strategies in adolescent idiopathic scoliosis: preliminary analysis of 3 patients and 6 scenarios. AB - Since the introduction of modern multi-segmental instrumentation systems, disagreement exists about the appropriate instrumentation strategies for the "optimal" correction of scoliotic deformities, and the difference between alternative scenarios is difficult to predict a priori. The purpose of this study is to evaluate the effect of different instrumentation strategies using a computer assisted surgery simulator (S3). We obtained from 32 experienced Fellows of the Scoliosis Research Society and members of the Spinal Deformities Study Group the detailed preoperative planning for three AIS patients with Lenke curve types 1A, 3A and 5C. Their scenarios were individually simulated using a computer model implemented in a spine surgery simulator (S3). The resulting Cobb angles varied for the 3 cases (e.g.: main thoracic: 6-17 degrees; 16-29 degrees; 16-30 degrees). The variability of correction remained important when sub-classifying the results according to the instrumentation strategies: A- "Pedicle Screws Constructs"; B- "Hooks Constructs"; C- "Hybrid Constructs". But overall, the average correction was better in group A (71%) than in groups B (55%) and C (54%). For the first time the effect of various instrumentation strategies can be assessed preoperatively thanks to S3. A large variability of instrumentation strategies exist within experienced surgeons and these produce rather different results. This study also questions the criteria for optimal configuration and standards to objectively design the best surgical construct. PMID- 17108445 TI - Objectives for correction and related instrumentation strategies in scoliosis surgery for Lenke curve types 2, 3 and 5. AB - A recent study revealed a large variability among a group of 32 spine surgeons in the pre-operative instrumentation planning for the same 5 AIS patients. It is hypothesized that this variability may be attributed to different objectives for correction. In this new study we analyzed the objectives of correction and the related instrumentation strategies for three different Lenke curve types. Nine experienced surgeons from the Spinal Deformity Study Group were surveyed and asked to assess 11 different geometric parameters describing the spinal deformities for three different Lenke curve types (2, 3 and 5) according to their importance for an optimal 3D correction. These same 9 surgeons were asked to provide their preferred posterior instrumentation planning for three patients with the same curve types. Statistical analyses included: median, interquartile range IQR and Wilcoxon non parametric test. There was an overall agreement that sagittal and coronal balances were the most important parameters for an optimal correction. All other parameters were highly variable depending on the curve type. Mobility was more important for the Lenke curve types 3 and 5 than for type 2 (p<0.032). A comparative analysis based on the coronal curves (Cobb) and the number of unfused vertebrae revealed a significant difference (p<0.025) between the correction objectives of the surgeons and their posterior instrumentation planning. In the three curves types analyzed, there is a large variability in scoliosis correction objectives, which is surgeon and curve-type dependent. There is a disagreement between the correction objectives and the instrumentation strategies. Optimal configuration of surgical instrumentation remains a controversial topic. PMID- 17108446 TI - Correction of the angle of trunk inclination utilizing apical wiring techniques and concave rib osteotomy in adolescent idiopathic thoracic scoliosis. AB - Improving the angle of trunk inclination (ATI) is important for achieving optimum clinical deformity correction in adolescent idiopathic scoliosis. Three concave apical implant techniques were reviewed in combination with the use of concave rib osteotomies. Rib osteotomy improved the ATI correction significantly when using apical wiring techniques. PMID- 17108447 TI - The use of thoracoplasty in the surgical treatment of idiopathic scoliosis. AB - INTRODUCTION: Rib prominence on the convex side results from vertebral rotation. The cosmetic deformity of the back in scoliosis is only partially corrected by operations on the spine itself, whilst costoplasty addresses the problem directly, and improves the cosmesis. PURPOSE: Our experience in convex and concave side thoracoplasty is discussed. PATIENTS AND METHODS: The selection of the patients for thoracoplasty was done primarily taking into consideration the cosmetic disturbance of the rib hump and the consequences to the psychism of the patient. A total of 35 scoliotic patients (32 females and 3 males) with mean age 18.8 years underwent thoracoplasty in combination with posterior spinal fusion. In 23 patients (3 males and 20 females) convex side thoracoplasty (rib resection at the site of the hump) was done as a first stage procedure (18 patients) or a second stage procedure (5 patients). In the patients with spinal fusion at the same time, the resected ribs were used as bone graft. 12 female patients were treated with concave side thoracoplasty (osteotomies of the medial part of the ribs and elevation of the ribs on the instrumentation rod) as a first stage procedure combined with spinal fusion, while in one 22 female patient both side thoracoplasty was done as a second stage procedure. CONCLUSION: Either form of thoracoplasty was an effective and impressive way to improve the patient's appearance although it was not possible to quantify the results. The complications that were presented viz. 4 haemopneumothorax, 2 pneumothorax, 2 pneumonia) were treated successfully. PMID- 17108448 TI - Validity and reliability of SRSI and SF-36 in Japanese patients with scoliosis. AB - We have examined the validity and reliability of Japanese SRS-24 and SF-36 in patients with scoliosis. 278 patients with scoliosis were recruited. Their average age was 24.8 years (range 17-84 years) and 83% of patients had idiopathic curves. The major coronal curve averaged 43 degrees (range 10 degrees -114 degrees). Questionnaires surveyed were SRS-24 and SRS-22, and SF-36. Only domains applicable to untreated patients were analyzed. Scale level analysis was evaluated by the ceiling and floor effect. The Cronbach alpha and item-scale correlations were calculated as representatives of internal consistency reliability and item internal consistency, respectively. Most domains of SF-36 had large ceiling effects. Both the SRS questionnaires had limited ceiling/floor effects. SRS-22 had a better internal consistency than SRS-24. Low item-scale correlations (<0.25) were found in question 14, 15, and 18 of SRS-22. The Japanese version of SRS-22 was the most useful measure for patients with scoliosis. However, some questions have low item-scale correlations, lowering reliability of the Function domain. PMID- 17108449 TI - Current methods for management of scoliosis. Treatment or malpractice? AB - Bracing and spine instrumentation are the universally used methods for the treatment of scoliosis used during the last half century. This now demands an objective evaluation of the long-term outcome of treatment in order to determine our contemporary attitude to these unique in medicine, time-resisting therapeutic paradigms. This is essential, with the need to report ambiguity of results of bracing and the accumulated evidence of high morbidity, with sometimes severe complications, of instrumentation. It is also mandatory with regard not only to the demands for the continuous development of scientific principles, based on effective methods for treatment with less side effects to the maximum benefit of the patient, but also from an ethical point of view in accordance with existing moral axioms and requirements for professional integrity. PMID- 17108450 TI - Deformity related stress in adolescents with AIS. AB - A new questionnaire (The Bad Sobernheim Stress Questionnaire BSSQ) has been developed to assess the psychological stress scoliosis patients develop as a consequence of their deformity. The aim of this study was to determine this stress level in a sample of adolescents with scoliosis. MATERIALS AND METHODS: 206 Patients were recruited to the study and completed a questionnaire (8 items). The average age was 15.7 years with an average Cobb angle of 35.8 degrees. All subjects returned a fully completed questionnaire. Plausibility of the results was assessed by examining for consistent responses to two plausibility questions. RESULTS: The average stress value was 19.97 / 24, which can be regarded as "low stress". The average stress value in the group meeting the plausibility threshold (n = 155) was 21 / 24 while the average stress value in the group not meeting the threshold (n = 51) was 15.9 / 24, regarded as "medium stress". This correlated negatively with the Cobb angle (r = -.54; p < 0,001). The Cobb angle / test value correlation was highest in thoracic curves (n = 87; r = -.49; p < 0,001) and lowest in double major curves (n = 46; r = -.27; p = 0,03). CONCLUSIONS: Adolescents with scoliosis have on average only "low stress" associated with their deformity. Not fulfilling a plausibility threshold seems to be a predictor for more stress associated with the scoliotic deformity. PMID- 17108451 TI - The influence of brace on quality of life of adolescents with idiopathic scoliosis. AB - Traditionally, the effectiveness of brace treatment on adolescents with IS is based on curve magnitude and to some extent on vertebral rotation and rib hump. QoL has been introduced in the recent years in order to evaluate the effectiveness of brace treatment. The aim of the study is to determine the influence of brace on quality of life (QoL) of adolescents with idiopathic scoliosis (IS).Thirty-six patients with a mean age of 13,9 (range 12-17) years old, a mean Cobb angle 28,2 degrees (range 19-38 degrees and a mean angle of trunk inclination (ATI) 7,8 degrees (range 4 degrees -17 degrees) who were treated conservatively with a modified Boston brace for a minimum of 2 years, filled the form of Brace Questionnaire (BrQ). BrQ is a validated, disease specific instrument, its score ranges from 20 to 100 and higher BrQ scores mean better quality of life. Correlations were determined by the Pearson correlation coefficient, with p<0.05 considered significant. Mean overall score of BrQ was 73,8 (SD 15. 8). Lower scores were observed in physical functioning (55,4, SD 15,9) and vitality (55, SD 25,9). School activity (98, SD 4,4) was affected less. The Cobb angle correlated significantly only with school activity (p<0,02). The ATI correlated significantly with social functioning (thoracolumbar ATI, p<0.038; lumbar ATI, p<0.035). There were no significant correlations between either Cobb angle or ATI with BrQ overall scores. Cobb angle and ATI significantly influenced school activity and social functioning respectively, but not general health perception, physical functioning, emotional functioning, vitality, bodily pain and self-esteem and aesthetics. PMID- 17108452 TI - Associations between quality-of-life and internal or external spinal deformity measurements in adolescent with idiopathic scoliosis (AIS). AB - In theory, quality-of-life measures should correlate with impairments such as spinal deformity. The goal was to determine the ability of the SRS-22 and surface topography measures to predict internal deformity. Data from 227 females with AIS were collected. Correlations and regression were used to predict Cobb angle or category. In subjects treated conservatively, the Cobb angle correlated with pain, self-image, satisfaction and total score. Only self-image correlated with surface topography. In subjects having had surgery, Cobb angle correlated with self-image, mental health, satisfaction and total scores. The cosmetic score was the only external deformity measure to correlate with the SRS-22. Function, self image and trunk-twist predicted subjects within 3 categories (Cobb <30 degrees , 30-50 degrees , >50 degrees) with 57% accuracy compared to 53% when using self image only. Accuracy in predicting subjects with curves smaller or larger than 50 degrees was 79% (using self-image, trunk-twist) or 72% (self-image only). Correlations between quality-of-life and deformity measures were low. However, it was possible to predict subjects within clinically meaningful categories of internal deformity using SRS-22 scores. PMID- 17108453 TI - Evaluation of torso shape and asymmetry associated with scoliosis. AB - This paper presents a technique for evaluating torso shape and asymmetry in scoliosis using indices of point-set data obtained from torso surface scans. Point-set data are the maximum curvature points of a surface scan and contain key shape information. The centroids, distributions and densities of point-set data are indices of shape and asymmetry. The mean-squared deviations of the centroids per-cross-section from the medial plane are indicative of asymmetry. The distribution and density of point-set data are indicative of shape. Evaluations using point-set data indices were 100% in accord with those based on the cosmetic score. Indices of point-set data have more degrees of freedom and were able to distinguish between scans that have similar cosmetic scores. PMID- 17108454 TI - Assessment of brace local action on vertebrae relative poses. AB - Bracing is a widely used treatment of scoliosis, but there is still no consensus about its actual effect. Previous studies were based on global descriptors of the spine shape (Cobb angle, plane of maximal deformity, etc.). We present a new method to analyze braces effects at a finer scale and to find which vertebral levels are significantly affected by this treatment. The proposed method compares a group of patients treated with a brace and a control group. The 3D spine geometry of the patients from the two groups was digitized on two separate occasions: with and without brace (first group) or two times without brace (control group). The modifications of the vertebrae relative poses (combination of relative translation and rotation between consecutive vertebrae) were then extracted from 3D reconstructions. Centrality and dispersion measures of the relative poses modifications were computed using a method that take into account the non-linearity of the rotation matrix. Then, finally, multivariate hypothesis tests were used to compare the centrality and dispersion of the two groups. The method was applied to 28 untreated scoliotic patients and 41 patients treated with a Boston brace. Significant differences (p<0.01) between the centrality and dispersion measures of the relative poses modifications were respectively found from T1 to T6 and from T8 to L1. Those significant differences concords with the back flattening effect and the spatially limited correction found in other studies; however the proposed method offers a more specific evaluation of the localization of those effects. PMID- 17108455 TI - Design of a minimally invasive non fusion device for the surgical management of scoliosis in the skeletally immature. AB - Current management of scoliosis in the skeletally immature focuses on prevention of curve progression. Progressive curves require surgical instrumentation without fusion with periodic lengthening of the construct, continuous bracing and ultimately fusion of the curve when control is lost at the adolescent growth spurt. This treatment regime has a major negative impact on the childs biopsychosocial profile and yields relatively poor longterm results. The aim of this study was to define the ideal characteristics of a growing rod and design a novel implant based on these characteristics. The method used was logical thought to define the desired implant characteristics and then design an implant which approximated the theoretical model as closely as possible. An ideal growing rod would grow with the spine, correct the deformity as growth occurred, leave no residual deformity at maturity, not require activity restriction/bracing, not require fusion at maturity and be compatible with a minimally invasive surgical technique. The ideal growing rod (IGR) is modular consisting of a number of units linked together which conform to the shape of the deformed spine. Each unit spans a disc space and consists of a telescopic rod with one end attached to the vertebra above and the other to the vertebra below. The rod is pushed apart by the force of growth but can only elongate in a predetermined direction resulting in a three dimensional realignment of the instrumented spinal segments with growth. In conclusion it is possible to design a growing rod with theoretical advantages over current constructs. PMID- 17108456 TI - Etiologic theories of idiopathic scoliosis: the breaking of bilateral symmetry in relation to left-right asymmetry of internal organs, right thoracic adolescent idiopathic scoliosis (AIS) and vertebrate evolution. AB - In the search to understand the etiology and pathogenesis of adolescent idiopathic scoliosis (AIS) some workers have focused on mechanisms initiated in embryonic life including a disturbance of bilateral (left-right or mirror-image) symmetry highly conserved in vertebrates. The normal external bilateral symmetry of vertebrates results from a default process involving mesodermal somites. The normal internal asymmetry of the heart, major blood vessels, lungs and gut with its glands is also highly conserved among vertebrates. It results from the breaking of the initial bilateral symmetry by a binary asymmetry switch mechanism producing asymmetric gene expression around the embryonic node and/or in the lateral plate mesoderm. In the mouse this switch occurs during gastrulation by cilia driving a leftward flow of fluid and morphogen(s) at the embryonic node (nodal flow) that favors precursors of the heart, great vessels and viscera on the left. Based on the non-random laterality of thoracic AIS curves, the hypothesis is suggested that an anomaly of the binary asymmetry switch explains the excess of right/left thoracic AIS. Some support for this hypothesis is the prevalence of right and left scoliosis curve laterality associated with situs inversus. There is recent evidence that vertebrates within their bilateralised shell retain an archaic left-right asymmetric visceral body organization evident in thoracic and abdominal organs. PMID- 17108457 TI - Etiologic theories of idiopathic scoliosis: enantiomorph disorder concept of bilateral symmetry, physeally-created growth conflicts and possible prevention. AB - The detection of anomalous extra-spinal left-right skeletal length asymmetries in the upper limbs, periapical ribs, ilia and lower limbs of subjects with adolescent idiopathic scoliosis (AIS) raises questions about skeletal bilateral symmetry of vertebrates in health and disorder, its origin and control. The vertebrate body plan externally has mirror-image bilateral symmetries that are highly conserved culminating in the adult form. The normal human body can be viewed as containing paired skeletal structures in the axial and appendicular skeleton as 1) separate left and right paired forms (eg long limb bones, ribs, ilia), and 2) united in paired forms (eg vertebrae, sternum, skull, mandible). Each of these separate and united pairs are mirror-image forms--enantiomorphs. Left-right asymmetries of growth plates (physes) may cause (1) in long bones length asymmetries, (2) within one or more vertebral physes putative growth conflict with distortion as deformity, and (3) between ribs and vertebrae putative growth conflict that triggers thoracic AIS suggesting preventive surgery on spine and ribs. There is evidence of a possible role for environmental factors in AIS development. Genes and the environment (nature/nurture) may interact pre- and/or post-natally to explain both the deformity of AIS and its association with widespread anomalous skeletal length asymmetries. If substantiated there may ultimately be a place for the prevention of AIS in some subjects. PMID- 17108458 TI - Bilateral intra-annular spinal compressive stresses in vivo. AB - The purpose of this pilot study was to determine if compressive stresses in the annulus of the intervertebral disc vary with activity in a quadruped and are affected by treatment with an implant. Pilot in vivo tests were conducted on skeletally immature domestic pigs (approved by IACUC). One pair of sensors was implanted within the annulus of T10-11, and the second pair at T8-9. A staple was them implanted across the right side of T8-9. Wires were routed subcutaneously and exited at the dorsal cervical region. Sensor signals were acquired before and after staple implantation, post-operatively during normal activities, and biweekly under anesthesia. After 8 weeks, spines were harvested and imaged. Early results from 2 sensors during walking and sitting, post-op day 5, clearly showed cyclic stresses during gait. Stresses were attenuated at the stapled vertebra compared to the unstapled vertebra. PMID- 17108459 TI - Postural differences in the shoulder girdle during normal locomotion in treadmill vs. over ground walking. AB - In a clinical setting, most musculoskeletal assessment related to gait is undertaken using a treadmill which is additionally widely used during gait and neurological rehabilitation. Although previous studies have reported the range of motion of gait characteristics during running, there is a paucity of information on normative walking data applicable to clinical assessment. Movements and posture of the shoulder girdle is an important indication of back and upper extremity function. While studies indicate that shoulder girdle kinematics are changed during shoulder dysfunction there is little information on the postural relationship between the shoulder and pelvis. The present investigation aims to create a normative database for kinematics during treadmill walking by examining the differences between the treadmill and over ground walking patterns. Additionally, the research will also assess shoulder girdle posture and to establish its relationship with the pelvic complex which will contribute to further understanding scoliotic posture and movement. 14 normal subjects walked over ground at a self-selected speed followed by walking on the treadmill at a speed matched to each subject's respective average over-ground speed. Three dimensional kinematic data was captured using a passive marker based motion analysis system (Vicon Peak, UK). Angular and temporal kinematic parameters were estimated. The results indicate differences in angular kinematics between over ground and treadmill locomotion. These differences should be considered when treadmill kinematics are used for clinical evaluation as opposed to over ground ambulation is scoliosis and other conditions. PMID- 17108460 TI - Balance lower limb loads and 3D spine modifications after total hip joint replacement: effects of leg length discrepancy correction. AB - Following total hip joint replacement (THJR), the durability of a prosthesis is limited by: wearing of frictional surfaces and loosening and migration of the prosthesis-cement-bone system. Literature review witnesses biomechanical studies focused mainly/only on hip functional state while none of them approached leg length discrepancy (LLD), posture unbalancing or spine related problems after THJR. Conversely, these latter could be critical elements for surgery and rehabilitation success, given the possible induction of asymmetric loading patterns. This study presents the results obtained by using a recently proposed methodology, to measure 3D subject posture balance and spine morphology and to evaluate its usefulness in individual therapy tuning/follow up. 3D subject's posture has been measured by means of 3D opto-electronic device, force platform and baropodography. 90 subjects after THJR have been included in this study. The subjects have been evaluated in two different epochs: 3 weeks after surgical intervention and after 3 months. 77/90 patients presented a LLD, pelvic obliquity and posture unbalancing. More than 90% of this group showed an overall postural re-balancing induced by the use of simple underfoot wedge. 70/77 patients needed wedge under the healthy side showing the surgical intervention produced a leg lengthening. 60/90 (52 LLD) patients underwent up to now to control and patients who wore the suggested wedge (63.4%) presented an improvement over all the considered quantitative parameters. Patients who wore a shorter than suggested wedge (23.1%), or that did not wear the suggested wedge (13.5%) presented a moderate or significant worsening of their postural balancing respectively. PMID- 17108461 TI - Biomechanical modeling of anterior spine instrumentation in AIS. AB - This study is part of a larger project regarding the development of a Spine Surgery Simulator (S3), which has shown good results for posterior instrumentation surgeries. The aim was to develop a biomechanical model for the anterior instrumentation of the scoliotic spine. A biomechanical model using flexible mechanism was developed and surgical manoeuvres (instrumentation, rod installation and compression) were reproduced. Validation of the model was done by comparing the results for the instrumented part of the spine to the post operative data (analytical Cobb angles in the frontal and sagittal planes, plane of maximum deformity, etc.). To date, surgeries of four patients operated by thoracotomy were reproduced. Preliminary results show that anterior instrumentation of the scoliotic spine can be adequately modelled using pre operative geometric data and using mechanical properties from literature. Once validated with a larger sample of cases, the anterior instrumentation model could be implemented into S3 and used by orthopaedic surgeons to test various instrumentation strategies in virtual reality before performing the actual surgery. PMID- 17108462 TI - Towards an automatic classification of spinal curves from x-ray images. AB - The objective of this research was to describe a multi-resolution technique for segmentation of the spinal curve on postero-anterior radiograph. Global features from the radiographs were first identified. From this approximate region, local regions of the spine curvature were iteratively filtered using diffusion and shock filters to enhance the boundaries of each vertebral body from radiographic noise. The spine curve going through the centroid of the vertebral bodies was successively identified for clinical classification of spinal deformities. PMID- 17108463 TI - Prevalence of subtle cardiac electrical abnormalities in children with idiopathic scoliosis. AB - We aimed to assess the prevalence of ECG abnormalities in children with idiopathic scoliosis (IS). 77 Girls and 13 boys, aged 7 to 18 years (15 +/- 3), including 12 with thoracic scoliosis (mean Cobbe angle 33 degrees), 4 with lumbar scoliosis (29 degrees), 12 with thoracolumbar (27 degrees), and 62 with double major scoliosis (31 degrees) entered the study. They were grouped as follows: 20 children <14 years of age (A), 35 from 14 to 16 (B), and 35 >16 years of age (C). Routine ECG was recorded and analysed automatically (GE, CASE v.4.1). Several ECG indices were further analysed. Abnormal values were considered if they exceeded upper normal limit (>95 percentile). RESULTS: Abnormalities were found in 66 patients (73%) independent of age. QRS duration gt;90 ms was observed in 40 patients (44%), right axis deviation in 28 (31%), left axis deviation in 2 (2%). The Rsr'(V1-2 ) pattern was noticed in 25 patients (28%). Ventricular gradient gt;60 degrees was found in 11 patients (12%). In patients with normal ECG there was a normal leftward axis rotation with age (-10 degrees +/-18 degrees in gr.A, 5 degrees +/-17 degrees in gr.B and +2 degrees +/-22 degrees in gr.C, difference from median), whereas reversed trend (rightward deviation with age) characterized patients with ECG abnormalities (-7 degrees +/-30 degrees in gr.A, +9 degrees +/ 19 degrees in gr.B and +19 degrees +/-2 degrees in gr.C, p<0.001). CONCLUSION: In children with idiopathic scoliosis, subtle ECG abnormalities are frequent. Abnormal trend of rightward QRS axis deviation with age suggests cardiac involvement in natural history of scoliosis and requires more depth cardiac evaluation. PMID- 17108464 TI - A biomechanical study of L5-S1 low-grade isthmic spondylolisthesis using a personalized finite element model. AB - The pars interarticularis lesions in isthmic spondylolisthesis result generally from mechanical stresses in the neural arch due to repetitive overload during daily activities or to trunk imbalance resulting from spino-pelvic morphology. The L5-pelvis junction of a low-grade isthmic spondylolisthesis patient was modeled using a personalized finite element model to investigate the biomechanical behavior of the L5-S1 motion segment. Stress distribution and facet contact pressure in the altered segment were analyzed under 500 N simulating the gravitational load. High von Mises stresses were located in the pars interarticularis, in the pedicle, and in the outer region of the annulus. A high contact pressure was obtained at the facet surfaces. Influenced by the pelvic morphology, the inclination of L5-S1 junction affects the resulting shear forces and may play a crucial role in spondylolisthesis development. PMID- 17108465 TI - Double crush syndrome evaluation in the median nerve in clinical, radiological and electrophysiological examination. AB - Double crush syndrome (DCS) was first described by Upton and McComas who proposed that focal compression of an axon often occurs at more than one level. The aim of the study was to support the hypothesis of DCS of the median nerve on the basis of available diagnostic methods. 30 patients (25 F and 5 M aged 33-73, mean 54.6+/-8.2 years) with coexisting carpal tunnel syndrome (CTS) and cervical radiculopathy (CR) were examined. Control group included 40 healthy volunteers (27 F and 13 M aged 17-82, mean 43.1+/-11 years). Medical evaluation comprised clinical examination, X-ray and MR imaging of the cervical spine, electroneurography (ENG) with F-wave and somatosensory evoked potentials (mSEPs) of median nerves. In clinical examination 96.6% of patients suffered from cervical spine pain and nocturnal paresthesies of at least one hand. Muscular atrophy was present in 43.3% in the proximal and in 70% in the distal part of the upper extremity. 30.3% of patients presented with a thoracic scoliosis. On X-ray examination, all patients showed cervical discopathy, mostly C5-C6 (70%) and C6 C7 (53.3%). Using MR investigation, the narrowing of intervertebral foramina was present in 81.25% and narrowing of vertebral canal in 37.5%. On ENG all patients presented with CTS, bilaterally in 73.3%. The F wave was abnormal in 73.3% and mSEPs in 66.7% of patients. Coincidence of MR and mSEPs in view of lateralization was observed in 71.4%. Results supported the DSC hypothesis. DCS evaluation requires both structural and functional diagnosis of peripheral neurones using MRI and electrophysiological examination. PMID- 17108466 TI - Handedness and spinal deformity. AB - Biological lateralisation is clearly manifest in scoliosis, yet its relevance is unclear. Goldberg et al. (Spine. 15(2):61-64. 1990) found an association between curve pattern and hand-preference in a screened population, but no increase in sinistrality. Milenkovic et al, (European Journal of Epidemiology, 19:969 972,2004) concluded left-handedness was a risk factor in a screened group. The database was reassessed to determine whether clinically significant scoliosis was associated with sinistrality or differed from the population norm of 10%. Patients attending the scoliosis clinic were asked their preferred writing hand. 1,636 patients were identified with complete data. Overall, left handedness occurred in 11.5%, greater than the general population (p=0.04) Left hand preference was found to be increased in boys with infantile idiopathic scoliosis and in girls with infantile, juvenile, congenital and syndromic scoliosis, but was reduced in girls with adolescent idiopathic scoliosis. Scoliosis lateralisation was random in infantile and congenital scoliosis, while left curve patterns were decreased in girls with juvenile idiopathic scoliosis and increased in boys with syndromic scoliosis. Curve pattern and handedness correlated in asymmetry in boys and girls and in girls with radiologically confirmed adolescent idiopathic scoliosis, but not in any other type. This study cannot confirm findings of left-handedness as a risk factor for spinal deformity. Its incidence is reduced in girls with adolescent idiopathic scoliosis, and the increased sinistrality in infantile scoliosis is not a new finding (Rauterberg & Tonnis Ger. Z.Orthop. 109(14):676-689. 1971). Lateralisation is undoubtedly a factor in scoliosis, but does not have a simple causal relationship, probably deriving from the underlying scoliotic process, rather than contributing to it. PMID- 17108467 TI - Surface topography and vectors: a new measure for the three dimensional quantification of scoliotic deformity. AB - The monitoring of spinal deformity uses many techniques: clinical history and physical examination for patient status, radiography for precise spinal delineation and Cobb angle, topography to quantify cosmesis and to approximate the Cobb angle. Experience with a system based on Raster photography has shown that adequate correlation with the Cobb angle is achieved, but that the relationship between spinal curvature and cosmetic effect is not simple. A measure was developed to quantify the asymmetry of the back, making it available to statistical analysis, without expressing it in terms of the Cobb angle or referring to trunk balance or rotation. The calculation expresses symmetry about the median saggittal plane (first thoracic vertebra to natal cleft), reflecting the right half onto the left and measuring the three-dimensional displacement between corresponding fixed points on the trunk. Tolerance limits were calculated and correlation with Cobb angles using routine scans was analysed. There were statistically significant correlations between the Cobb angle and all vectors except the middle antero-posterior. All vectors correlated with each other, except again for the middle Z or anteroposterior which correlated only with the middle and lowermost sets. Applied to natural history and to surgical outcome, this new parameter provides a different quantification of back shape which can be used both for patient assessment and monitoring, for the evaluation of the cosmetic (as opposed to the radiological) effect of treatment, and for aetiology and natural history studies. PMID- 17108468 TI - Quality of life after surgical decompression of lumbar spinal stenosis with and without instrumentation. AB - The aim is to evaluate the influence on quality of life of surgical decompression with and without instrumentation in lumbar spinal stenosis. Twenty three patients, (16 women, 7 men) with a mean age of 62,8 years old (range 44-80) who underwent a surgical decompression for lumbar spinal stenosis filled the SF-36 questionnaire pre- and postoperatively, during the follow up period which was at a mean value of 42 months (range 6 to 50 months). Spinal stenosis was degenerative in 18 patients and as a consequence of spondylolisthisis in 5. In 15 patients decompression and fusion using instrumentation (group I) was performed and in 8 patients only decompression was performed (group II). Statistical analysis was carried out using the Wilcoxon Signed Rank Test. In group I, the domains that evidenced statistical significant improvement were bodily pain (p<0,041), general health (p<0,042), vitality (p<0,042), social functioning (p<0,043), and mental health (p<0,042). Not any specific domain in group II showed a statistical significant improvement postoperatively. Comparing the two groups overall SF-36 score, a statistical significant improvement was noted for group I (p<0,001) and for group II (p<0,017). The statistical significance of improvement was stronger in patients of group I than group II. Surgical decompression for lumbar spinal stenosis reduces pain and restores significantly physical and mental health. Decompression and instrumentation presents superior results in patients' quality of life when compared to patients that single decompression was performed. PMID- 17108469 TI - Consequence of paraspinal muscle after spinal fusion: an experimental study. AB - Posterior lumbar spinal fusion is a common surgery of spine. The parapsinal muscular dysfunction was postulated to be due to injured paraspinal muscle inflicted by the surgery. To better understand the muscle dysfunction after spinal fusion, this descriptive study evaluated electrophysiology and histology changes in paraspinal muscle of rabbits with and without fusion. Three New Zealand white rabbits underwent 2-level posterior spinal fusion with instrumentation. Anothor 3 rabbits underwent surgical exposure alone and served as sham control. Pre-operative and follow-up electromyography (EMG) and histological assessment were performed in a 6-month interval. All fusion achieved solid union judged by post-mortem examination. Root mean square (RMS) of EMG showed decrease in both fusion and control group immediately after surgery, but the control group recovered to pre-operative value at 6 months follow-up. At 6 months after operation, the fusion group showed lower RMS in fused region than adjacent region. In term of median frequency (MF) changes, the control group did not show obvious difference between pre- and post-operative period. However, fusion group showed obvious decrease of MF in fusion region, but increase in lower adjacent region. Histology reviewed fatty infiltration in fusion region at 6 months after spinal fusion. In addition, the muscle fiber size presented smaller in fusion region than that of pre-operative status. These results would demonstrate the lower activities and muscle disuse atrophy of paraspinal muscles in the fusion region. While, higher muscular activities were found in the lower adjacent region, which may indicate muscle hypertrophy. PMID- 17108470 TI - The development of osteoblasts from stem cells to supplement fusion of the spine during surgery for AIS. AB - Surgical correction in severe cases of AIS is often hampered by insufficient autograft bone to facilitate the fusion. The development of other sources of bone generating cells would greatly enhance the surgical. Bone marrow derived stem cells were harvested from femoral reaming during total hip arthroplasty for the purpose of differentiating into osteoblasts. Stem cells were isolated from the marrow and successfully differentiated into three cell lines (osteoblasts, chondrocytes and adipocytes) to confirm multilineage potential. Osteoblasts were developed from the stem cells and demonstrated the ability to be cultured to possibly provide a source of bone generating cells to augment surgical fusions. It is anticipated that the addition of osteoblasts created from stem cells (combined with appropriate matrix) will have significant influence on the success of AIS surgery through improvement of bone fusion. PMID- 17108471 TI - Recent observations in the biomechanical etiology of so-called idiopathic scoliosis. New classification of spinal deformity--I-st, II-nd and III-rd etiopathological groups. AB - The article examines the biomechanical etiology of so-called idiopathic scoliosis (AIS). It describes I-st, II-nd and III-rd etiopathological groups (epg) of spine deformity which were developed during the years 2001/2004/2005. All children with so-called idiopathic scoliosis had an abduction contracture of the right hip, often connected with a flexion and external rotation contracture. In other cases we found only limited range of adduction of the right hip in comparison to the left hip. We maintain that children with this real abduction contracture of the right hip constitute the first etiopathological group of the development of scoliosis (I-st epg). This group has an "S" double shaped scoliosis with the rib hump on the right. Other patients, with only limited adduction of right hip in comparison to the left hip, constitute the second etiopathological group of development of scoliosis (II-nd epg). This group has a "C" shaped lumbar, sacro lumbar or lumbo-thoracic left convex scoliosis. The third etiopathological group (III-rd epg) shows either no or a minimal curve on X-ray with either no rib hump or a very minor one but have a "stiffness of spine". Such patients have problems with sporting activities and, as adults, the spinal stiffness leads to considerable "back pain". The right hip structural abduction contracture, or the differences in adduction, is connected with the "syndrome of contractures" in neonates and babies described by many authors and in depth by Mau. How does scoliosis develop? Our explanation is as follows. Asymmetry of movement of the hips during gait provokes asymmetry of loading and asymmetry of growth of both sides - left and right - and the gradual development of scoliosis. In I-st epg, the scoliosis is a secondary compensation for deformities in the pelvis and spine. The II-nd epg is linked to a permanent standing posture maintained on a free right leg during the first years of life. The III-rd epg comprises of patients from the boarder groups of I-st and II-nd epg. This classification establishes a clear therapeutic approach to every etiopathological group of scoliosis and allows for the possibility of introduction of causative prophylaxis. PMID- 17108472 TI - Pre-processing range data for the analysis of torso shape and symmetry of scoliosis patients. AB - This paper presents a procedure for pre-processing and reconstructing surface scans comprising an interactive technique for cropping stray points and extremities. The procedure involves three stages: cross-sectioning and clipping, hole-filling and sub-sampling, and surface re-generation. The accuracy of the reconstruction obtained was assessed by creating different patterns of holes and stray points on 30 models of the torso and pre-processing the models using the proposed procedure and existing procedures based on Bezier interpolation and Moving Least Squares (MLS) interpolation. Results obtained indicate that the proposed procedure was at least as good as the better of Bezier and MLS interpolation for all the models tested, particularly outperforming both procedures when holes account for up to 5% of the surface. Its accuracy of reconstruction ranged from 90-100% compared to 80-100% for Bezier and 50-100% for MLS. This work is a crucial step in developing techniques for understanding and assessing changes in torso shape and symmetry from torso surface scans of scoliosis patients. PMID- 17108473 TI - Adjacent segment disc pressures following two-level cervical disc replacement versus simulated anterior cervical fusion. AB - Anterior cervical fusion (ACF) has been shown to alter the biomechanics of adjacent segments of the cervical spine. The goal of total disc replacement is to address pathology at a given disc with minimal disruption of the operated or adjacent segments. This study compares the pressure within discs adjacent to either a two-level simulated ACDF or a two-level total disc replacement with the ProDisc-C. A special automated motion testing apparatus was constructed. Four fresh cadaveric cervical spine specimens were affixed to the test stand and tested in flexion and extension under specific loads. Intradiscal, miniature strain-gauge-based transducers were placed in the discs above and below the "treated" levels. The specimens were then tested in flexion and extension. Pressure and overall angular displacement were measured. In the most extreme and highest quality specimen the difference at C3/C4 registered 800 kPa and the difference at C6/C7 registered 50 kPa. This same quality specimen treated with the ProDisc reached a flexion angle at much lower moments, 24.3 degrees at 5 N-m, when compared to the the SACF 12.2 degrees at 8.6 N-m. Therefore, the moment needed to achieve 15 degrees of flexion with the SACF treatment was 5.5 N-m and the ProDisc treatment was only 2.9 N-m. This initial data would indicate that adjacent level discs experience substantially lower pressure after two-level disc replacement when compared to two-level SACF. Additional testing to further support these observations is ongoing. PMID- 17108474 TI - The effect of instrumentation with different mechanical properties on the pig spine during growth. AB - The effect off mechanically altered bone on spinal growth using instrumentations with different mechanical properties is quantified through the use of experimental tests. Eight spine segments from three female pigs weighing 60 90 lb were subjected to a continuous tensile force. The load (0 - 400 N) was applied using a MTS Alliance RT/50 machine and the resulting extension recorded using an extensometer. Displacements between the screws were measured in control without implant, metal plate system, and spring system, respectively. It is shown that the addition of an implant will increase the stiffness of the spine, which will affect the extension of the spine and hence hinder growth in the spine. PMID- 17108475 TI - Improving brace wear with active brace system. AB - There is considerable controversy regarding the effectiveness of brace treatment for patients with adolescent idiopathic scoliosis (AIS). Researchers believe that to be effective, patients must wear their braces as prescribed including both compliance and tightness. Compliance is how much time the brace is worn relative to the prescribed time. Brace tightness is usually prescribed by orthotists during brace fitting session. Asking the patient or examining the brace for wear are the most common methods to evaluate the brace usage. A low powered microcomputer system was developed to monitor and maintain loads exerted by braces used to treat children with spinal deformities during daily living. This system records brace usage information and helps patients to wear their brace at the prescribed tightness. Laboratory tests have been performed and six patients have used the system for four weeks. The patients reported that the system helped them to wear the brace properly. The time that the patients wore the braces at the prescribed tightness level increased from 48+/-16% during the monitor period (first 2 weeks) to 63+/-18% during the automatic adjustment period (last 2 weeks). PMID- 17108476 TI - 3D back shape in healthy young adults: an inter-rater and intra-rater reliability study. AB - Whilst postural evaluation of spinal dysfunction is routine in physiotherapy practice, objective measurements are rarely undertaken due to the scarcity of reliable low cost assessment tools. Warren et al [12] found high intrarater reliability but inter-rater reliability has not as yet been evaluated. The purpose of this study was to assess both the intra-rater and inter-rater reliability of the Middlesbrough Integrated Digital Assessment system (MIDAS). METHODS: A convenience sample of twenty-five healthy University of Teesside students was recruited for the study. One rater palpated fifteen key landmarks on each subjects back. Each of three raters took two measurements on each subject in a standardized upright posture. Rater order was randomized to minimize data recording bias. X (medio-lateral), Y (antero-posterior) and Z (height) landmark positions were recorded via a computer interface. DATA ANALYSIS: Intraclass Correlation Coefficients (ICC 2,1) were used to analyse data using SPSS v13. RESULTS: Both intra-rater agreement (mean ICCs - rater 1 r= 0.970, rater 2 r= 0.965 and rater 3 r= 0.965, p<0.001) and inter-rater agreement (mean ICCs r = 0.967, p<0.001 was very high between repeated measures and between markers. Error values for the z-axis (height) were lowest. CONCLUSIONS: The system demonstrated both high inter-rater and intra-rater reliability. Before the system is used on a clinical population, data output needs to be converted from raw format to a clinically applicable format. Work is currently being undertaken to develop an interactive visual display and control for postural sway. PMID- 17108477 TI - Comparison of isometric trunk rotational strength of adolescents with idiopathic scoliosis to healthy adolescents. AB - Trunk rotational strength asymmetry has been suggested in adolescents with idiopathic scoliosis (IS), but is unconfirmed. The sitting isometric trunk rotational strength, at neutral and 18 degrees or 36 degrees of right or left pre rotation, of a group of healthy adolescent females (CG), n=12, is compared with a group of female adolescents with IS (ISG), n= 14. Torque values were normalized to lean body weight. There is a significant weakness when rotating towards the concavity found in patients with AIS at the 36 lo (p 0.07 marginal), 18 lo (p<0.03) and neutral positions (p<0.02), with no side strength asymmetry found in a cohort of healthy adolescents without AIS. PMID- 17108478 TI - Intra-operative spinal load and displacement monitoring: towards a better understanding of scoliosis correction mechanics. AB - Surgical correction of scoliosis reduces deformation and improves overall function and esthetics. Understanding and monitoring of mechanics during scoliosis surgery is an invaluable tool to optimize correction without compromising patient safety. Our objective was to use intra-operative monitoring tools to study how spinal load and displacement relates to obtained correction and chosen instrumentation. Instrumented pedicle screws, a "gripper" and active markers were developed. Instrumented pedicle screws provided three-dimensional forces at the screw-vertebra interface while the instrumented "gripper" measured the force and the rotation applied by the surgeon to the rod rotator. Vertebral displacement was monitored with light-emitting diodes and motion capture technology. These instruments were used successfully with 16 scoliosis subjects. Analysis of applied force, displacement, and curve flexibility influence on correction percentage is the long term goal. Raw results for instrumented screws and gripper showed that recorded force decreased with respect to percentage of correction obtained. Measured force increased with respect to the pre-surgical Cobb angle while percentage of correction obtained decreased as pre-surgical Cobb angle increased. Active marker results showed three-dimensional vertebral rotation and translation during correction, with axial rotation and caudal cranial translation having the greatest magnitudes. Using greater correction forces does not necessarily result in an increased correction; flexibility and Cobb angle also play a role in the mechanics of correction. Further data collection will provide better understanding of the interconnected role between these factors helping complete the description of surgery mechanics. PMID- 17108479 TI - A controlled prospective study on the efficacy of SEAS.02 exercises in preparation to bracing for idiopathic scoliosis. AB - The Lyon school has proposed a preparation to brace wearing through an intensive mobilization in order to obtain a better reduction of the braced scoliotic curve. Our aim was to verify this hypothesis. DESIGN: A prospective controlled study on consecutive patients having idiopathic scoliosis with brace management. OUTCOME: Results after 5 months of brace wearing were reviewed by radiographic examination without the brace. TREATMENT: SEAS Group exercises according to the protocol SEAS.02 (Scientific Exercises Approach to Scoliosis, version 2002); CONT Group various type of exercises. Population. 110 patients (34 females), 13.5+/-2.4 years, 31.1 degrees +/-11.1 degrees Cobb (degrees C), 14.4 degrees +/-6.0 degrees Bunnell (degrees B). All parameters improved at follow-up in both groups. SEAS had better results than CONT for degrees C. Clinical results (variations of at least 5 degrees C and 2 degrees B) were better in SEAS than CONT. This study proves the efficacy of SEAS.02 exercises preparatory for bracing. Bracing demonstrated its short term efficacy. PMID- 17108480 TI - A controlled prospective study on the efficacy of SEAS.02 exercises in preventing progression and bracing in mild idiopathic scoliosis. AB - There is low evidence on the possible efficacy of exercises to treat idiopathic scoliosis, graded as C by the existing Italian Guidelines. Our aim was to verify if exercises quality has an effect on results. DESIGN: Prospective controlled study on idiopathic scoliosis patients that performed only exercises to avoid progression. TREATMENT: SEAS Group make exercises according to the protocol SEAS.02 (Scientific Exercises Approach to Scoliosis, version 2002). The CONT Group performed exercises at a local structure according to different protocols preferred by the treating therapists. Population. SEAS: 48 patients (37 females), 12.5+/-2.2 years, 15.1 degrees +/-5.7 degrees Cobb (degrees C), 9.0 degrees +/ 3.3 degrees Bunnell (degrees B). The difference in the number of braced patients within the first year has been almost statistically significant (P=0.07): 1 in SEAS vs. 5 in CONT. Cobb degrees improved with treatment (P<0.05) only in the SEAS group. Clinical results (variation of at least 5 degrees C or 2 degrees B) were better in SEAS than CONT. Not all exercises for scoliosis have the same efficacy: this study proves the short term efficacy of SEAS.02 when compared to usual care. PMID- 17108481 TI - An innovative diagnostic procedure of vertebral deformities without exposure to X rays. AB - The objective of the study was to compare standard manual X-ray measurements of vertebral deformities and values obtained from the Ortelius 800. 52 Patients (41 females and 11 males: mean age 20.35 years) with adolescent vertebral deformities, was studied. The patients were evaluated with standard radiographic views and the Ortelius 800. The parameters considered for the comparison were the angles of scoliosis and kyphosis and the values of global axial deformity, shoulder asymmetry and pelvic tilt. We also evaluated the modification of pelvic/shoulder angle after surgery (this parameter allows to evaluate vertebral rotation and can be derived only from the Ortelius 800). This study allowed us to conclude that there is a perfect agreement between measurements with the Ortelius 800 and those resulting from standard x-rays. The system has also proved to have the capability of quantifying changes in vertebral rotation. PMID- 17108482 TI - Semi-automatic detection of scoliotic rib borders using chest radiographs. AB - Stereoradiography is a well known technique to obtain 3D reconstructions of the rib cage. However, clinical applications are limited by the associated 2D rib detection method. Either this detection is widely supervised and time-consuming for the user, or it is fully automatic and not accurate enough for proper 3D reconstruction or clinical indices extraction. To address these issues, we propose a novel, semi-automated technique for detecting scoliotic rib borders in PA-0 degrees and PA-20 degrees chest X-ray images, using a modified edge following approach. The novelty consists in following multiple promising edges simultaneously. Detections are initiated from starting points (input by the user) along the upper and lower rib edges and the final rib border is obtained by finding the most parallel pair among the detected edges. Promising results show the superiority of this approach over classical rib detection in terms of accuracy. Moreover, the proposed method is of great relevancy in the scoliotic context since scoliotic ribs present very few shape priors, due to their irregularities, and hence, standard rib detection techniques become unsuitable. PMID- 17108483 TI - Brachial plexus palsy associated with halo traction before posterior correction in severe scoliosis. AB - OBJECTIVE: To retrospectively analyse clinical features and related factors of brachial plexus palsy associated with halo traction before posterior correction in severe scoliosis. METHOD: 300 Cases of severe scoliosis performed with halo traction before posterior correction were considered with 7 cases suffering from brachial plexus palsy (2 males and 5 females). The average age was 14 years (range, 9-19 years). The average Cobb angle was 110 degrees (range, 90 degrees - 135 degrees); Diagnoses were idiopathic scoliosis (1), congenital scoliosis (3), and neuromuscular scoliosis (3). Halo-gravity traction was used in 3 cases preoperatively; and Halo-femoral traction used in 4 cases postoperatively (anterior release 2 cases, anterior epiphyseal arrest 1 case, combined anterior and posterior release 1 case). RESULTS: Traction was used for an average of 3.5 weeks before spinal fusion (range, 2-6 weeks) for these 7 patients. The average traction weight was 8 kg; the average traction weight was 19 % ( range 13-26%) of the average body weight (40.2 kg). The mean stature was 175 cm; all the 7 patients had a long and thin body configuration. Duration between brachial plexus paralysis and detection was 1 to 3 hours. All the 7 patients suffered different degree from numbness of ulnaris of the hand and antebrachium. Median nerve palsy was found in 3 cases, ulnar nerve paralysis was found in 4 cases. Complete nerve functional restoration had been achieved by the end of three months after rehabilitation training, drug treatment were adopted. CONCLUSION: Brachial plexus palsy associated with halo traction in severe scoliosis is related to the weight of traction, body type and patient-pathology status. If the symptoms are promptly detected with rehabilitation training and appropriate drug treatment adopted, complete nerve functional restoration can be achieved. PMID- 17108484 TI - Radiologic presentations in relation to curve severity in scoliosis associated with syringomyelia. AB - The purpose of the study was to further explore the radiological presentations of scoliosis associated with syringomyelia in relation to curve severity. 87 children and adolescents were divided into three groups: Group I (10 degrees 60 degrees). Curves were classified into typical and atypical patterns in the coronal plane and the sagittal profile measured. Cerebellar tonsillar descent or syrinx patterns in relation to curve severity and the frequency of atypical curves were also investigated. The frequency of atypical curve patterns from Group 1 to 2 was 46.2%, 45.2%, and 40.7% respectively. 65.3% patients with typical curve patterns had atypical features in all of the three groups. There was a significant difference of kyphotic angle among the three groups. Both the degree of cerebellar tonsillar descent and syrinx patterns had no correlation with the curve severity or the frequency of atypical curves. These results show that radiographic presentations, including atypical curve patterns, atypical features in typical curve patterns, and a normal to hyperkyphotic thoracic spine may suggest the need for preoperative MRI. There is no evidence to suggest that the degree of cerebellar tonsillar descent and syrinx patterns have an effect on the progress of scoliosis and the frequency of atypical curves. PMID- 17108485 TI - Evaluating similarity metrics in an image matching tool for image guided spine surgery. AB - This paper presents a comparison study of the effect of three similarity measures (mutual information, normalized mutual information, and mean squared error on the edges of the input image) in an image matching tool that can be used in image guided spine surgery. Using 3D rotational x-rays and magnetic resonance images of a spine phantom, it was found that the similarity measures had similar effect. Therefore, experiments with other datasets are needed before making conclusions about the suitability of these similarity metrics for image guided surgery. PMID- 17108486 TI - Right convex thoracic female adolescent scoliosis in the light of the thoracospinal concept. AB - The most common form of idiopathic scoliosis affects perimenarcheal girls who show either a primary or secondary right convex thoracic curve. The patient displays signs of sympathetic dysfunction, morphological and histochemical abnormalities of muscle fibres and platelets and a persisting osteoporosis. Mechanisms that have been proposed for the causation and the development of these features are supported by the evidence in results of studies into the thoracospinal concept of the etiopathogenesis of this form of scoliosis. The interrelation of these late integrated signs and symptoms suggests that this particular form of IS is a disparate clinical nosological entity rather than a mere orthopedic deformity of the spine. Known characteristics of the infantile and this form of adolescent scoliosis further suggest that the juvenile form is an intermediate rather than a separate group and that the use of the term "idiopathic" is, therefore, obsolete. Deciphering the etiopathogenesis of the pathological complex of the right convex female adolescent scoliosis demands research from new standpoints which demands fresh approaches. PMID- 17108487 TI - Instrumentation loosening and material of implants as predisposal factors for late postoperative infections in operated idiopathic scoliosis. AB - INTRODUCTION: Instrumentation loosening and metal corrosion are predisposal factors under investigation for late Postoperative infections. PURPOSE OF THE STUDY: To investigate the contribution of the instrumentation material (stainless steel versus titanium implants) and the mechanical loosening in the development of late postoperative spinal infection. PATIENTS AND METHODS: The first group of patients involves 50 idiopathic scoliotic patients who were treated with first generation posterior stainless steel spinal segmental multihook instrumentation. The minimum post operative follow up was 4 years. Five patients presented with late infections 1 to 5 years post operatively. Removal of instrumentation was the effective solution to this problem. Common intraoperative findings were some degree of instrumentation loosening and corrosion. The second group involves 40 idiopathic scoliotic patients who were treated with newer generation posterior titanium spinal segmental multihook-multiscrew instrumentation system. More extensive use of pedicle screws was performed to the second group resulting in a more stable mechanical construct. Follow up ranged from 2 to 5 years. None of those patients presented late postoperative infection or any evidence of instrumentation loosening or failure. CONCLUSION: We believe that newer multihook multiscrew titanium spinal instrumentation systems have smaller incidence of late postoperative infections because they provide a more stable construct (pedicle screws) with fewer tendencies for micro motion or failure, and they may give the advantage of greater bone adhesion on the implant resulting in the production of thinner biofilm, thus decreasing the chances of infection. PMID- 17108488 TI - Variations of the position of the cerebellar tonsil in adolescent idiopathic scoliosis with severe curves: a MRI study. AB - This study is an investigation into the position of the cerebellar tonsils in AIS with severe curves and any relationship it has with age, sex, curve severity and curve pattern. Sagittal magnetic resonance imaging (MRI) of hindbrain was performed on both of 205 AIS patients with a Cobb angle greater than 40 degrees and 86 healthy controls. The position of the cerebellar tonsil relative to the line connecting the basion and opisthion (BO line) was measured. In AIS and controls, the mean position of the cerebellar tonsil was 0.9 and 2.9 mm above the BO line, respectively. The incidence of tonsillar ectopia in AIS was found to be significantly higher than controls (34.5% versus 5.8%, p<0.001). No significant correlations were found between the position of the cerebellar tonsil with age or sex in AIS and controls. It was shown the position of the cerebellar tonsil was not significantly different among AIS patients with deferent curve severity. However, a significant lower incidence of tonsillar ectopia (p=0.049) was found in patients with lumbar curves when compared to those with thoracic or thoracolumbar curves. In conclusion, there was a relatively lower position of the cerebellar tonsil together with a significant higher incidence of tonsil ectopia in AIS patients. There was a trend that tonsillar ectopia was more often in thoracic or thoraco-lumbar curves, suggesting that a lower position of the cerebellar tonsil may play an important role in the etiopathogenesis of AIS. PMID- 17108489 TI - Correction of scoliosis using segmental pedicle screw instrumentation versus hybrid constructs with hooks and screws. AB - Eighteen consecutive patients with scoliosis surgically treated by posterior correction and fusion were recruited. The major coronal curve averaged 65.8 degrees (range 51 degrees -87 degrees ). Eight patients were treated by hybrid constructs and four of them had anterior release before posterior surgery. Ten patients were treated by posterior correction and fusion alone with pedicle screw instrumentation. Flexibility was assessed by radiographs using the active bending and the fulcrum bending techniques. Two correction indices were not statistically different between the two groups. However, there was a tendency towards better correction in the screw group: mean surgical correction was 62.6+/-11.8% in the hook group and 71.6+/-12.3% in the screw group (p=0.07). Comparable pre- and post operative radiographic results may indicate the advantages of posterior-only segmental pedicle screw instrumentation over hybrid constructs. PMID- 17108490 TI - Changes of three-dimensional back contour following posterior fusion for idiopathic scoliosis. AB - Twenty-nine patients received both pre and post operative Quantec evaluation (Raster stereophotograph) and spinal radiography. Mean age at surgery was 13.9 years and mean age at follow-up was 15.9 year old. Patients have been followed at least 6 months post-surgically, ranging from 6 months to 5 years. The study found that except for an improvement of Cobb angles (from 49.64 degrees to 24.81 degrees in the thoracic; from 56.08 degrees to 20.08 degrees in the thoracolumbar), corrections of rib hump (from 16.88 to 11.40 vs. normal range 0 to 10.) and truncal asymmetry (from 36.97 to 21.92 vs. normal range 5 to 20) are critical factors in successful spinal surgery. PMID- 17108491 TI - First results of pain treatment in scoliosis patients using a sagittal realignment brace. AB - For adult scoliosis patients with chronic pain bracing is indicated before spinal surgery is considered. The treatment with the "sagittal realignment brace" (physio-logic brace) has clearly improved chronic low back pain in a short test period. Thus we started a prospective study to estimate also medium term effects. MATERIAL AND METHODS: 29 Patients (Cobb angle 37 degrees ; age 41 years) have been treated with the physio-logic brace for an average of 7,5 Months (SD 5,6). Before brace prescription the brace action was simulated using the sagittal realignment test. Pain intensity was recorded before brace treatment, within the 1(st) week after brace adjustment and at follow-up using the Roland and Morris VRS. Period of daily wearing time as well as total wearing time / day was left up to the patients. RESULTS: Pain intensity was high (3,38) before treatment (scale 0-5) and low (1,28) in the brace after adjustment. The differences where highly significant in the Wilcoxon test (z=-4,29; p < 0,0001). After follow-up the pain intensity was moderate to severe (2,69; ns). There was a clear reduction of pain severity, however in most of the cases the compliance has been lost during the follow-up period. Only 7 Patients continued to wear the brace for a considerable time (> 4 hrs.). CONCLUSIONS: The brace action of the sagittal realignment brace leads to promising short-term effects. A better compliance might be achieved by prescribing the total wearing time / day and though a better efficiency of pain reduction in the long-term. PMID- 17108492 TI - Brace treatment of spinal claudication in an adult with lumbar scoliosis--a case report. AB - Although spinal claudication may arise from narrowing of the spinal canal, not all patients with narrowing develop symptoms. The reason why some patients develop symptomatic stenosis and others do not is still unknown. Therefore, the term lumbar spinal stenosis refers to a clinical syndrome of lower extremity pain caused by mechanical compression on the neural elements or their blood supply. Some studies have shown effectiveness of brace treatment with a common supportive LSO. At our Centre the sagittal realignment brace is used for the treatment of chronic low back pain. This is a lumbar lordosing brace theoretically leading to a reduction of the volume in the spinal canal. However a patient with a significant increase in walking distance due to the application of this brace will be presented here. MATERIAL AND METHOD: A 47 year old woman with a 55 degrees lumbar scoliosis, 30 degrees upper lumbar kyphosis and with highest pain levels under medication (Durogesic 25 mg, Ibuprofen 800, Mirtazapin 15 mg) has been treated with a sagittal realignment brace. Self reported walking distance was at around 800 steps before the pain appeared unbearable (since 5 years). Self reported walking distance was recorded (Patients counts) in the brace 2 days and 10 days after adjustment. RESULTS: Walking distance increased to 8000 steps after 2 days and to 12000 after 10 days while pain intensity decreased only one point in the VRS, however without any further medication. CONCLUSIONS: In contrary to current hypotheses about the aetiology of spinal claudication augmentation of lordosis may lead to a significant improvement of symptoms associated with spinal stenosis and lumbar scoliosis. PMID- 17108493 TI - Brace treatment of spinal claudication in an adolescent with a grade IV spondylosisthesis--a case report. AB - Although spinal claudication may arise from spondylolisthesis, little information exists about successful conservative treatment of this condition. However there are studies describing pain reduction due to physiotherapy and bracing. Significant improvement of walking distance and pain intensity in an adolescent with scoliosis, spondylolisthesis and spinal claudication while wearing a delordosing spondylogic brace is presented here. MATERIAL AND METHOD: A 14 year old girl with a 25 degrees thoracic scoliosis (2 years postmenarchial), grade IV spondylolisthesis and spinal claudication underwent treatment with a delordosing spondylogic brace. Walking distance without brace was at around 300 steps before intolerable pain appeared. Self reported walking distance was recorded in the brace 14 days after adjustment. RESULTS: Walking distance increased to an unlimited number of steps after 14 days while pain intensity decreased three points in the VRS. However, no correction effect of the orthosis on the degree of slippage was found. CONCLUSIONS: Although there is evidence that pain in patients with spondylolithesis can be reduced using exercises and bracing in mild to moderate symptomatic cases, this case demonstrates that bracing can also improve signs and symptoms of spinal claudication in patients with spondylolisthesis of higher degrees. A prospective case series study seems desirable. PMID- 17108494 TI - ADL based scoliosis rehabilitation--the key to an improvement of time-efficiency? AB - A new ADL (Activities of Daily Living) approach in scoliosis rehabilitation has been designed. The ADL approach uses Side-Shift, physiologic(R), 3D-ADL exercises and Schroth exercises according to current standard. It was compared to an exercise based scoliosis rehabilitation that mainly uses Schroth exercises and as an add-on the training of ADL. The ADL approach in practice seems to be easier to teach and the treatment needs fewer theoretical modules leading to a better time efficiency. Aim of this study was to test whether the gain in time-efficiency is at the loss of outcome. MATERIALS AND METHODS: We studied 13 patients having had a 2 weeks ADL based rehabilitation (ABR) and compared their clinical outcome parameters (surface topograhy & Scoliometer angle) to a group of 13 diagnosis-, age-, sex-, Cobb-angle and curve pattern-matched controls having a 4 weeks programme of exercise based rehabilitation (EBR) only. Average age in the study group was 15 years and average Cobb-angle 39 degrees. RESULTS: Lateral deviation in the study group already after 2 weeks of ABR decreased 2mm (Formetric). Lateral deviation in the control group (EBR) decreased 1,9 mm. The changes were not significant. Thoracic as well as lumbar Scoliometer angle decreased highly significantly in both groups (ABR: 1,8 degrees thoracic, 2,3 degrees lumbar; EBR: 2,1 degrees thoracic, 2,6 degrees lumbar). The differences of outcome between the two groups were not significant. CONCLUSIONS: ABR (2-week programme) seems to provide similar results as EBR (4-week programme). ABR seems to provide a better time efficiency, however a prospective controlled study with a larger sample of patients is desirable before final conclusions can be drawn. PMID- 17108495 TI - Lower degree of mineralization found in cortical bone of adolescent idiopathic scoliosis (AIS). AB - A lowered bone mineral density (BMD) was observed in trabecular bone in a significant number of AIS patients. The present study aims at investigating whether lower BMD is a systemic phenomenon, which would also be present in the cortical component of long bone. SUBJECTS AND METHODS: 78 AIS girls (age:15 18y.o) with either moderate (Cobb: 20-40) or severe (Cobb: >40) curve and 44 age matched controls were recruited. The BMD of the distal region and the mid-shaft of radius were measured with a multi-layer peripheral quantitative computed tomography (pQCT). The trabecular bone and cortical bone BMD and the morphology of mid-shaft were compared. RESULTS: Both trabecular and cortical BMD in severe AIS group was significantly lower than the control by 8.7% and 1.7% (p<0.05 for both), respectively. However, the cortical bone area of the mid-shaft did not show any differences from the normal control subjects. DISCUSSION: This study demonstrates a systemic low BMD including the cortical bone. It is suggested that AIS girls may have disturbance in mineralization and ossification during peripubertal growth. PMID- 17108496 TI - Gamma subunits expression of junctional acetylcholine receptor of paraspinal muscles in scoliosis associated with syringomyelia. AB - To investigate the denervation of paravertebral muscles in scoliosis associated with syringomyelia via detecting the subunit expression of junctional acetylcholine receptors. METHODS: All the cases were divided into three groups: Group A consisted of 20 patients with scoliosis associated with syringomyelia, Group B included 10 adolescents with idiopathic scoliosis, and Group C was 10 cases without scoliosis. Bilateral biopsy of the paraspinal muscles was performed during scheduled spinal surgery. The mRNA expression of junctional acetylcholine receptor (AchR) subunits were detected by RT-PCR method. RESULTS: 13 Patient (65%) in Group A had a positive expression of the mRNA for the gamma subunit. The positive rate of gamma subunit expression was 40% on the convex side of the curve, and 25% on the convex side. The gamma subunit expression was absent from all samples in Group B. In Group C, only one patient had the gamma subunit expression on the left side. CONCLUSION: The gamma subunit expression of the junctional acetylcholine receptor is changed in scoliosis associated with syringomyelia, suggesting the presence of denervation in paravertebral muscles. The denervation of paravertebral muscles may play an role in the pathogenesis of scoliosis. PMID- 17108497 TI - Superior mesenteric artery syndrome following scoliosis surgery: its risk indicators and treatment strategy. AB - From July 1997 to October 2003, 640 patients with adolescent scoliosis who had undergone surgical treatment were evaluated prospectively, 7 patients of whom suffered from SMAS post-operatively. The height of the 7 patients who developed SMAS was above the mean of sex- and age-matched normal population, with the height percentile ranging from 5% to 50%. On the contrary, their weight was below the mean of the normal population with a weight percentile ranging from 5% to 25%. Among the 7 patients, 4 had a thoracic hyperkyphosis ranging from 55 degrees to 88 degrees (average 72 degrees ) and 2 had a thoracolumbar kyphosis of 25 degrees and 32 degrees respectively. The 7 patients were all treated with fasting, antiemetic medication and intravenous fluids, a nasogastric tube was passed and aspirations commenced. Reduction or suspense of traction was adopted in three patients who had SMAS during Halo-femoral traction after anterior release of scoliosis. All the patients recovered completely with no sequelae. PMID- 17108498 TI - The young person's guide to biomedical informatics. AB - In a retrospective review, a parallel is drawn between the challenges by which a research department in biomedical informatics is confronted and those of a symphony orchestra. In both areas, different disciplines and different groups of instruments can be discerned. The importance of mastering one's instrument and the harmony between the team members is stressed. The conductor has to stimulate the individual players so that they can all have a successful career. Competition between orchestras and performance assessments determine survival and success. A record of refereed publications is crucial for continued existence. Conclusions are that biomedical informatics is typically multidisciplinary, that hypotheses underlying research should be carefully formulated, that the time from research to application may easily take 20 years or more, that mutual trust and knowing each other's competences is essential for success, that a good leader gives enough room to all team members to develop their careers, and that the outcomes of assessment studies are related to the quality of publications. PMID- 17108499 TI - Reflections on biomedical informatics: from cybernetics to genomic medicine and nanomedicine. AB - Expanding on our previous analysis of Biomedical Informatics (BMI), the present perspective ranges from cybernetics to nanomedicine, based on its scientific, historical, philosophical, theoretical, experimental, and technological aspects as they affect systems developments, simulation and modelling, education, and the impact on healthcare. We then suggest that BMI is still searching for strong basic scientific principles around which it can crystallize. As -omic biological knowledge increasingly impacts the future of medicine, ubiquitous computing and informatics become even more essential, not only for the technological infrastructure, but as a part of the scientific enterprise itself. The Virtual Physiological Human and investigations into nanomedicine will surely produce yet more unpredictable opportunities, leading to significant changes in biomedical research and practice. As a discipline involved in making such advances possible, BMI is likely to need to re-define itself and extend its research horizons to meet the new challenges. PMID- 17108500 TI - Meeting the challenges--the role of medical informatics in an ageing society. AB - The objective of this paper is to identify trends and new technological developments that appear due to an ageing society and to relate them to current research in the field of medical informatics. A survey of the current literature reveals that recent technological advances have been made in the fields of "telecare and home-monitoring", "smart homes and robotics" and "health information systems and knowledge management". Innovative technologies such as wearable devices, bio- and environmental sensors and mobile, humanoid robots do already exist and ambient assistant living environments are being created for an ageing society. However, those technologies have to be adapted to older people's self-care processes and coping strategies, and to support new ways of healthcare delivery. Medical informatics can support this process by providing the necessary information infrastructure, contribute to standardisation, interoperability and security issues and provide modelling and simulation techniques for educational purposes. Research fields of increasing importance with regard to an ageing society are, moreover, the fields of knowledge management, ubiquitous computing and human-computer interaction. PMID- 17108501 TI - Progress and challenges of ubiquitous informatics in health care. AB - Ubiquitous informatics in health care can be seen as its pervasively presence everywhere, at home as well as in office and patient room, for various purposes such as patient follow up, health care professional training, aid to decision making and to public health management. Its worldwide rapid extension could only happen in relation to major progresses such as overall availability of personal computers, diffusion through the worldwide web as well as coverage of almost all fields of medicine. Challenges include a profound change in patient-physician relationship, a reform in health care management and financial methods, as well as the need to identify uniquely all healthcare partners, while respecting confidentiality and private life. PMID- 17108502 TI - Facilitating interdisciplinary design specification of "smart" homes for aging in place. AB - "Smart homes" are defined as residences equipped with sensors and other advanced technology applications that enhance residents' independence and can be used for aging in place. The objective of this study is to determine design specifications for smart residences as defined by professional groups involved both in care delivery to senior citizens and development of devices and technologies to support aging. We assessed the importance of specific devices and sensors and their advantages and disadvantages as perceived by the interdisciplinary expert team. This work lays the ground for the implementation of smart home residencies and confirms that only an interdisciplinary design approach can address all the technical, clinical and human factors related challenges associated with home based technologies that support aging. Our findings indicate that the use of adaptive technology that can be installed in the home environment has the potential to not only support but also empower individual senior users. PMID- 17108503 TI - Rehabilitation after stroke using virtual reality, haptics (force feedback) and telemedicine. AB - We have constructed a haptic immersive workbench to be placed in the patients' home for daily adjusted rehabilitation. We also propose a system for Internet based connection and communication between patients and between patients and a clinical rehabilitation center and clinical assessment/evaluation centers. The benefits of a system for rehabilitation after stroke, based on VR, Haptics and Telemedicine should be: increased quality of life, lesser isolation, feeling more secure, fewer tiring transportations, more frequent exercising, better compliance to training, lower cost for transportation. The long term recovery for a larger group of patients with motor impairments is presently under evaluation. PMID- 17108504 TI - A distributed shared data space for personal health systems. AB - Ubiquitous computing is a promising paradigm to support health care outside traditional care institutes. Sensor-based systems may continuously collect data on a person's health status and context, and provide immediate feedback or contact a remote physician. This paper presents a novel programming model to facilitate the development of such systems. The model, which has been inspired by tuple spaces, offers robustness for ad hoc mobile environments and explicit support for data streams. PMID- 17108505 TI - Ubiquitous technologies in health: new challenges of opportunity, expectation, and responsibility. AB - In spite of their name, 'ubiquitous' technologies are not yet ubiquitous in the true sense of the word, but rather are 'novel', being at the research, pilot, and selective use stages. In future, the proliferation in types of application, the major increase in cases and data volumes, and above all the dependence on ubiquitous technologies will raise practical, ethical, and liability issues. Equally significantly, it will require health service redesign, including new response services. Health informaticians need to be active in stimulating consideration of all these issues, as part of both social and professional responsibility. PMID- 17108506 TI - Visualisation and interaction design solutions to address specific demands in shared home care. AB - When care professionals from different organisations are involved in patient care, their different views on the care process may not be meaningfully integrated. OBJECTIVE: To use visualisation and interaction design solutions addressing the specific demands of shared care in order to support a collaborative work process. METHODS: Participatory design, comprising interdisciplinary seminar series with real users and iterative prototyping, was applied. RESULTS: A set of interaction and visualisation design solutions to address care professionals' requirements in shared home care is presented, introducing support for identifying origin of information, holistic presentation of information, user group specific visualisation, avoiding cognitive overload, coordination of work and planning, and quick overviews. The design solutions are implemented in an integrated virtual health record system supporting cooperation and coordination in shared home care for the elderly. The described requirements are, however, generalized to comprise all shared care work. CONCLUSION: The presented design considerations allow healthcare professionals in different organizations to share patient data on mobile devices. Visualization and interaction design facilitates specific work situations and assists in handling specific demands in shared care. The user interface is adapted to different user groups with similar yet distinct needs. Consequently different views supporting cooperative work and presenting shared information in holistic overviews are developed. PMID- 17108507 TI - Safety portal: the safest goes through the air--ubiquitous high-risk reminders bridging out the patient safety in emergency department. AB - Under the safety net of hospital-based patient safety informatics (PSI) system, RFID is build to adapt emergency department safety needs and the process improvement activities. We implemented the RFID framework as the electronic patient identifier into the process improvement of ED workflow; it defined as the safety portal to introduce PSI safety features on the real-time basis. METHODS: Since 2004, we were applying RFID technology into a 200 daily visits emergency department of the regional medical center in Taipei, Taiwan. We then developed wireless web-based RFID safety portal to implement the real-time safety reminders such as the laboratory and radiology reports to the physician who can make decision promptly to the patients in the ED. RESULTS: Under the ED safety portal, the diagnosis time for physicians to make clinical decision is largely reduced nearly 40 percent due to the safety enhancement RFID system. ED physician who could actively receive patient's updated clinical data to make clinical decision via web-based informatics system reasonably decreases. CONCLUSIONS: The effectiveness of RFID system not only enhance patient identification during ED process but combine ED safety net which providing needed data for ED physician and staff who visit ED patients with valuable real-time data on time. Patient safety on ED can be clearly improved from the embracing modern technology and build up a patient centered ED environment. PMID- 17108508 TI - Highly automated documentation for mobile medical services. AB - Mobile Medical Services, such as Home Care and EMS (Emergency Medical Services) are to the general public probably most visible part of public health care. A great amount of expectations are placed on the quality of care given by these units. Sometimes providing this care is very intensive and all available attention has to be placed on the patient. However, documenting the treatment is very valuable for the treatment of the patient later on. In this paper we present a system that automates many tasks in documenting the treatment. Furthermore, our system is capable of producing a far more detailed documentation that has been available before. This makes reliable research of mobile medical care possible and opens new possibilities in educating paramedics and nurses. PMID- 17108509 TI - Development of a system supporting patient supervision and treatment in contemporary home-care: status report. AB - The emerging amalgamation of informatics, communication technologies, and entertainment electronics in the field of Biomedical Technology combined to, first, the increase in length of the mean life expectancy, and, second, the hospitalization cost avalanche, will facilitate gradually the development of a new Hi-Tec home-care environment. We have developed a home-computer based system, addressing crucial aspects of the development of contemporary home-care that comprises of: First, the employment of low-cost commercially available components, supporting home-care patient's well-being observation, including eventually vital-signs monitoring. Second, software means for the processing, the evaluation, and the targeted transmission of the acquired health-data. Third, software tools for the planning, the documentation, and the management of the corresponding home-care case. The present paper constitutes a progress report of the ongoing development efforts. PMID- 17108510 TI - Fuzzy Hidden Markov Models: a new approach in multiple sequence alignment. AB - This paper proposes a novel method for aligning multiple genomic or proteomic sequences using a fuzzyfied Hidden Markov Model (HMM). HMMs are known to provide compelling performance among multiple sequence alignment (MSA) algorithms, yet their stochastic nature does not help them cope with the existing dependence among the sequence elements. Fuzzy HMMs are a novel type of HMMs based on fuzzy sets and fuzzy integrals which generalizes the classical stochastic HMM, by relaxing its independence assumptions. In this paper, the fuzzy HMM model for MSA is mathematically defined. New fuzzy algorithms are described for building and training fuzzy HMMs, as well as for their use in aligning multiple sequences. Fuzzy HMMs can also increase the model capability of aligning multiple sequences mainly in terms of computation time. Modeling the multiple sequence alignment procedure with fuzzy HMMs can yield a robust and time-effective solution that can be widely used in bioinformatics in various applications, such as protein classification, phylogenetic analysis and gene prediction, among others. PMID- 17108511 TI - Modelling epidermal homeostasis as an approach for clinical bioinformatics. AB - Modelling in systems biology currently lacks clinical applications. As a possible approach leading to clinical relevance the modelling of tissue homeostasis is proposed. As an example a model of epidermal homeostasis is presented which reproduces central morphological and kinetic characteristics of epidermal tissue. Each individual cell is modelled as an agent. The tissue arises as an emergent phenomenon from the interactions of agents. Each agent's behaviour is qualitatively modelled by a simple differentiation state-flow program. Epithelialisation under the influence of parameters concerning stem-cell location is briefly demonstrated. PMID- 17108512 TI - Sifting abstracts from Medline and evaluating their relevance to molecular biology. AB - The most important knowledge in the area of biology currently consists of raw text documents. Bibliographic databases of biomedical articles can be searched, but an efficient procedure should evaluate the relevance of documents to biology. In genetics, this challenge is even trickier, because of the lack of consistency in genes' naming tradition. We aim to define a good approach for collecting relevant abstracts for biology and for studied species and genes. Our approach relies on defining best queries, detecting and filtering best sources. PMID- 17108514 TI - Inferring gene expression networks via static and dynamic data integration. AB - This paper presents a novel approach for the extraction of gene regulatory networks from DNA microarray data. The approach is characterized by the integration of data coming from static and dynamic experiments, exploiting also prior knowledge on the biological process under analysis. A starting network topology is built by analyzing gene expression data measured during knockout experiments. The analysis of time series expression profiles allows to derive the complete network structure and to learn a model of the gene expression dynamics: to this aim a genetic algorithm search coupled with a regression model of the gene interactions is exploited. The method has been applied to the reconstruction of a network of genes involved into the Saccharomyces Cerevisiae cell cycle. The proposed approach was able to reconstruct known relationships among genes and to provide meaningful biological results. PMID- 17108515 TI - Integrating medical and genomic data: a successful example for rare diseases. AB - The recent advances on genomics and proteomics research bring up a significant grow on the information that is publicly available. However, navigating through genetic and bioinformatics databases can be a too complex and unproductive task for a primary care physician. In this paper we present diseasecard, a web portal for rare disease that provides transparently to the user a virtually integration of distributed and heterogeneous information. PMID- 17108516 TI - HL7 RIM: an incoherent standard. AB - The Health Level 7 Reference Information Model (HL7 RIM) is lauded by its authors as 'the foundation of healthcare interoperability'. Yet even after some 10 years of development work, the RIM is still subject to a variety of logical and ontological flaws, which has placed severe obstacles in the way of those who are called upon to develop implementations. We offer evidence that these obstacles are insurmountable and that the time has come to abandon an unworkable paradigm. PMID- 17108517 TI - A reference model for clinical tumour documentation. AB - The definition of common system semantics is an explicit and generally accepted precondition for comparability and exchangeability of data from different systems. We have looked back on 15 years of experience with a data model that was developed in a co-operative effort by experts from various hospital cancer registries as the foundation of a new tumour documentation system (GTDS). The data model is based on the definition of a common basic data set for hospital cancer registries which is agreed by the German Association of Comprehensive Cancer Centres (ADT). This paper presents an "entity relationship" view of this model. Since data exchange among registries and with hospital or practice information systems is becoming increasingly important we describe our method to import data from such systems. We discuss the requirements that systems have to have for a most effective way of exchanging data with a hospital cancer registry. The most important feature is the possibility to associate disease phenomena and therapies with each other and with an entity that represents the tumour across encounters. The reference model we present respectively the requirements we propose for other communicating systems might also fit for other chronic diseases. PMID- 17108518 TI - Standards for enabling health informatics interoperability. AB - Most of industry countries are turning their healthcare system towards integrated care paradigms for improving quality, efficiency, and safety of patients' care. Integrated care has to be supported by extended communication and cooperation between the involved healthcare establishments' information systems. The required interoperability level goes beyond technical interoperability and simple data exchange as it has been started in the early world of electronic data exchange (EDI). For realising semantic interoperability, series of standards must be specified, implemented and enforced. The paper classifies standards for health information systems needed for enabling practical semantic interoperability. PMID- 17108519 TI - The HL7 reference information model under scrutiny. AB - The Health Level 7 (HL7) Reference Information Model (RIM) was once incepted as an object oriented information model to harmonize the definition of HL7 messages across different application domains. On the heels of the hugely successful HL7 version 2, version 3 and the RIM has received significant attention and credit and in turn is increasingly subjected to criticism. In this paper the authors, who are among the chief designers of the RIM, respond to the major points that have been raised against the RIM in the published literature. We find that much of the criticism is based on misunderstandings and differences in point of view. We wish to advance the dialogue in the hope that when we account for those differences, effective critique may lead to real improvements of the standard. PMID- 17108520 TI - Participation in the International Classification for Nursing Practice (ICNP) programme. AB - The International Council of Nurses (ICN) is a federation of 129 national nurses associations. The International Classification for Nursing Practice (ICNP) is a programme of the ICN. The purpose of this paper is to describe the development and maintenance processes of the ICNP Programme that are used to increase participation. These include processes by which the ICNP was and continues to be developed, tested, distributed and implemented worldwide, with emphasis on the current version, ICNP Version 1.0. The ICNP is a unified nursing language that facilitates cross-mapping among local terms and existing terminologies. ICNP conforms to current terminology standards and criteria, for example, ISO standards and HL7. The ICNP Alpha and Beta Versions documented the progress of concept validation and classification of nursing phenomena and interventions. The ICNP Beta 2 Version was a combinatorial terminology organized in two multi-axial structures representing nursing phenomena and nursing actions. The ICNP Version 1.0, launched in 2005, changed the relatively straight-forward multi-axial structure into a compositional terminology through the application of description logics using Web Ontology Language (OWL) within Protege, an ontology development environment. ICNP Version 1.0 is also represented in a multiaxial model (7-Axis) for nurses to compose nursing diagnosis, intervention and outcome statements. Language translations and clinical information systems applications are required to make the ICNP Version 1.0 available to nurses at the point of healthcare delivery. ICNP data collected in healthcare environments provide standardized terminology for nursing that allows comparison of nursing practice across health care settings, specialties and countries; facilitate data-based clinical and management decision making; and contribute to the development of guidelines and standards for best practices and optimal outcomes for patients, families and communities. PMID- 17108521 TI - A user-centred deployment process for ICT in health care teams--experiences from the OLD@HOME project. AB - OBJECTIVE: To present a user-centred method for introducing ICT in health care organisations, taking factors that influence acceptance into account. METHODS: User centred methods are used in combination with previous research regarding factors that affect user acceptance, in order to facilitate users' acceptance of new ICT tools. RESULTS: A method is presented that supports the introduction of ICT in team work. The method consists of three major steps; (1) the start-up seminar, (2) end user education and (3) continuous follow-up during the deployment phase. Important results of the start-up seminar are documentation of the users' expectations, and an agreement of ground rules that supports both the social norm factor and the users' perceived behavioural control. Education and follow-up also improve perceived behavioural control, and by involving super users perceived usefulness and ease of use can be improved through subjective norm. CONCLUSION: Key factors in the deployment process are; user participation, end user experience and education, and continuous follow-up of the process. PMID- 17108522 TI - Non-invasive light-weight integration engine for building EHR from autonomous distributed systems. AB - Pangea-LE is a message oriented light-weight integration engine, allowing concurrent access to clinical information from disperse and heterogeneous data sources. The engine extracts the information and serves it to the requester client applications in a flexible XML format. This XML response message can be formatted on demand by the appropriate XSL (Extensible Stylesheet Language) transformation in order to fit client application needs. In this article we present a real use case sample where Pangea-LE collects and generates "on the fly" a structured view of all the patient clinical information available in a healthcare organisation. This information is presented to healthcare professionals in an EHR (Electronic Health Record) viewer Web application with patient search and EHR browsing capabilities. Implantation in a real environment has been a notable success due to the non-invasive method which extremely respects the existing information systems. PMID- 17108523 TI - Research networks: can we use data from GPs' electronic health records? AB - As widely discussed in the literature, there are many potential scientific usages of data extracted from the primary care Electronic Health Records (EHR), such as quality of care, epidemiological or socio-economical studies. Yet, can we use the current available data in the EHR for such purposes? In this paper, our objective is to report on the preliminary findings of the Belgian ResoPrim project (2003 2005) to answer the question. We set up a semi-anonymous network involving 26 current practices (28 volunteer GPs), 3 different EHR software systems and two Trusted Third Parties. Based on a literature overview we identified 27 research questions to be answered using 50 indicators. The study design includes retrospective (2002-2004) and prospective (6 weeks) data collection processes around the theme of "Hypertension and cardiovascular risk factors". For some data sets, the data extraction was a full automatic procedure, for some others, the data extraction was related to an input from the GPs allowing some comparisons between both procedures. At this stage, we performed an extended descriptive analysis of our data. Retrospectively we collected data related to 42,217 patients and 203,128 contacts. Prospectively we collected data for 9,236 patients and 15,234 contacts. Our main findings are briefly presented and discussed in this paper. The most promising fields seem to be the Health Research Information Systems assessment and the quality of care studies. It is quite too soon to reach the expected theoretical benefits for epidemiologic and socio-economic studies, yet some progresses could be made in relation with the denominator issue. Based on our preliminary findings and hypotheses, further analyses are foreseen during the second phase of the project (2006-2007). PMID- 17108524 TI - Information technology in clinical research in rheumatology domain. AB - The development of a functional clinical database of rheumatic diseases represents an essential step in the process of acquiring the necessary epidemiological and other information on disorders under study. In 1999-2005 the Institute of Rheumatology in cooperation with the EuroMISE Center has developed the Clinical database/National Register of selected systemic inflammatory rheumatic diseases inclusive of a bank of sera and DNA. Aims of this phase of the pilot research were gathering clinical, laboratory, genetic, pharmaco-and socio economic data in a representative sample of patients with systemic lupus erythematosus, systemic sclerosis, polymyositis/dermatomyositis, mixed connective tissue disease; rheumatoid arthritis, juvenile chronic arthritis, ankylosing spondylitis, psoriatic arthritis and reactive arthritis. In 2002 the preset number of over 2000 registered patients had been achieved with collaboration of 34 territorial and 20 institutional rheumatologists in the whole covering the majority of the Czech Republic. Based on experiences gathered, the systems for other related studies are being developed and implemented using modern information technologies. PMID- 17108525 TI - Method for automatic escalation of access rights to the electronic health record. AB - In an emergency situation, it can be vital for rescuing personnel to have access to fragmented parts of patients Electronic Health Record (EHR) shared between patients and health care services. In such situations, can Spatial Role Based Access Control combined with measurements of vital sign parameters recorded from a wireless monitoring system used by the patient and patient's physiological situation be used to facilitate for medical personnel automatic access to parts of the EHR. PMID- 17108526 TI - The French proposal for a health identification number. AB - The French ministry of Health is setting up the personal medical record (PMR or DMP for Dossier Medical Personnel in French). This innovating tool is highly expected and will be extremely useful for the therapeutic follow-up as well as for epidemiological studies on which public health policies are based. Therefore the currently planned identifying process should prevent any epidemiological use of these data. Numerous scientific organisations have alerted government powers about the threat that this impairment represents, and they wish to promote some secure procedures that exist, which have already proved their efficiency at the national and international level. PMID- 17108527 TI - Can temporary paper-based patient records sensibly complete an electronic patient record? AB - OBJECTIVES: In the department of internal medicine of Heidelberg University Hospital (HUH), medical patient records are archived electronically. Since there are still paper-based external documents, a temporary patient record is maintained for these documents during the patient's stay. Afterwards, the paper based documents are scanned, indexed and integrated in the electronic patient record (EPR). To ensure process quality we evaluated quality and availability of scanned documents in the EPR. METHODS: Observation study, structured interviews, systematic quantitative before-after comparison. RESULTS: The workflow takes place according to the guidelines of HUH. Nevertheless, there are variations in the different wards which may influence the quality. Of 343 scanned documents about 90% showed no loss of information. Most of the documents with loss of information were ECG-curves. Four documents (1.2%) could not be found in the EPR. All documents were assigned to the correct patient and episode of care. The mean time from patient discharge to availability of scanned documents in the EPR system was 36 days. CONCLUSIONS: Due to external paper-based documents, a complete EPR is currently not possible. A temporary paper-based patient record in addition to the EPR is not an optimum procedure but feasible. The quality of the scanned, indexed and integrated documents in the EPR is high and the availability is sufficient. PMID- 17108528 TI - Ubiquitous information for ubiquitous computing: expressing clinical data sets with openEHR archetypes. AB - Ubiquitous computing requires ubiquitous access to information and knowledge. With the release of openEHR Version 1.0 there is a common model available to solve some of the problems related to accessing information and knowledge by improving semantic interoperability between clinical systems. Considerable work has been undertaken by various bodies to standardise Clinical Data Sets. Notwithstanding their value, several problems remain unsolved with Clinical Data Sets without the use of a common model underpinning them. This paper outlines these problems like incompatible basic data types and overlapping and incompatible definitions of clinical content. A solution to this based on openEHR archetypes is motivated and an approach to transform existing Clinical Data Sets into archetypes is presented. To avoid significant overlaps and unnecessary effort during archetype development, archetype development needs to be coordinated nationwide and beyond and also across the various health professions in a formalized process. PMID- 17108529 TI - Towards automatically generating graphical user interfaces from openEHR archetypes. AB - One of the main challenges in the field of Electronic Health Records (EHRs) is semantic interoperability. To utilise the full potential of interoperable EHR systems they have to be accepted by their users, the health care providers. Good Graphical User Interfaces (GUIs) that support customisation and data validation play a decisive role for user acceptance and data quality. This study investigates the use of openEHR archetypes to automatically generate coherent, customizable, data-validating GUIs. Using the Mozilla XML User Interface Language (XUL) a series of prototypes has been developed. The results show that the automatic generation of GUIs from openEHR archetypes is feasible in principle. Although XUL revealed some problems, the advantages of XML-based GUI languages are evident. PMID- 17108530 TI - Co-operation, interaction and information-sharing: from a paper-based Diabetes Pass to an electronic alternative. AB - Diabetes is one of the most challenging problems in the 21st century, whereby research showed that reducing the concentration of blood glucose is thought to prevent or reduce the long-term complications. To do so, a multidisciplinary approach is favourable. In Belgium, a revalidation programme for the diabetic patients concerning self-regulation was introduced followed by the introduction of the Diabetes Pass in March 2003 whereby some goals were stipulated. In IBBT COPLINTHO, a still ongoing project, an eHomeCare platform is implemented whereby the patient is the central actor. The analysis of the current paper-based Diabetes Pass revealed that the data can be easily extracted from the EHR. An electronic alternative for the Diabetes pass is proposed whereby the added values for the patient are underlined. Before implementing the electronic alternative, some research should still be done, but it is thought that all the actors involved--including the patient, could easily benefit from the electronic alternative. PMID- 17108531 TI - Personalized cardiovascular risk management linking SCORE and behaviour change to Web-based education. AB - The PULSE (Personalization Using Linkages of SCORE and behaviour change readiness to web-based Education) project objectives are to generate and evaluate a web based personalized educational intervention for the management of cardiovascular risk. The program is based on a patient profile generated by combining: (a) an electronic patient data capture template (DCT); (b) the Systematic COronary Risk Evaluation (SCORE) algorithm; and (c) a Stage of Change determination model. The DCT inherently contains a set of evidence-based parameters for patient description and disease evaluation. The patient's stage of behaviour change determines messages consistent with the individual's change processes, decisional balance, and self-efficacy. The interventions are designed to address both medical and psychosocial aspects of risk management and, as such, we combine staged lifestyle modification materials and non-staged messages based on Canadian clinical guidelines to motivate personal risk management. The personalization decision logic is represented in Medical Logic Modules implemented in Java. An intelligent interactive system generates the personally relevant materials and delivers the education to the patient via the Web. An evaluation study will be conducted to determine whether web-based personalized educational strategies exert favourable influence on patient's interest, knowledge, and perceived compliance to the suggested lifestyle modifications. PMID- 17108532 TI - Guarantying the quality of chemotherapy: from the order to the administration. AB - Ensuring the quality and security when prescribing drugs like chemotherapies is a complex task if one wants to cover the whole chain from the prescribing physician to the administrant nurse. At the University hospitals of Geneva, new applications covering the whole chain from the prescription up to and including the fabrication of the products have been developed in three phases and are being used in a production stage. In order to cover the "last yard" at the bed level, a fourth phase has been started with a pilot study based on labels containing RFID chips for preparations and for patients. The last phase will make use of all traceability data acquired from the prescription to the preparation to validate that the right product is administered to the right patient, and to record who is administrating it. PMID- 17108533 TI - Comprehensive management of the access to a component-based healthcare information system. AB - OBJECTIVE: to describe the key concepts and elements used to implement a comprehensive access management system to a distributed, component-based healthcare information system. METHODS: the a priori access is based on an institution-wide policy for access rights coupled to proximity process for the granting of such rights. Access rights are explicit and externalized from the information systems components. A posteriori control is based on a centralized, exhaustive journal of accesses to all components coupled to a decentralized verification process for suspicious accesses. RESULTS: the system has been operational for three years, initially used for the access to the computerized patient record components, and now extending to all the components of the hospital information system. The same architecture will be used for the development of the trans-institutional community health information network. PMID- 17108534 TI - Barriers integrating dedicated software for quality management in pain centres. AB - OBJECTIVES: Explore the feasibility of integrating a dedicated pain centre information system as part of a quality management network with a number of different Hospital Information Systems. MATERIAL & METHODS: A systematic approach integrating and implementing the system in 15 selected hospital organisations (a nationwide 15% non-random sample). RESULTS: Hospitals have widely varying policies on integration and implementation of additional clinically required 3rd party software. Financial and organisational constraints are considerable. Partial data integration could be realised in one third of the hospital organisations within the project timeframe. Linking with various types of Hospital Information Systems from the same or different vendors caused no technical difficulties. The total effort required, however, varies considerably; different versions of a HIS of the same vendor require substantial additional effort. IT departments hardly use standard rules to accept this type of systems, causing substantial increases in completion time for installation. CONCLUSIONS: Although it is feasible to integrate a local departmental system as required (download of general patient, provider and referrer data basically, and also upload of certain data) the workload of scaling to the national level is considered far beyond what is reasonable for a national quality management network for pain. Alternatives for recording and capturing data (which comply with the requirements of the national system) are currently being explored. PMID- 17108535 TI - On feasibility and benefits of patient care summaries based on claims data. AB - BACKGROUND: Electronic availability of health care claims data maintained by health insurance companies today is higher than the availability of clinical patient record data. OBJECTIVE: To explore feasibility of automatically generated patient care summaries based on claims data and its benefit for health care professionals (HCP), when no shared electronic health record is available. METHODS: Based on an existing claims data model for German health insurance companies, a transformation and presentation algorithm was developed. To determine the utility of the resulting summaries, a focus group session comprising HCP and insurance representatives was arranged. Properties of an information system architecture capable of providing summaries to HCP were specified. RESULTS: A set of valuable healthcare information, in particular clinical pathways, medication, and anamnesis, can be derived from claims data that fits into the ASTM specification of a Continuity of Care Record. The focus group assessed the potential benefit of the summaries as high. Major issues are partial incompleteness and a lack of timeliness due to delayed reimbursement procedures as well as privacy-preserving and practicable access methods. The specified system architecture uses web services and a web interface to provide the summaries in HL7 CDA format. An important insight was that only a timely electronic reimbursement process will lead to precise, current, and reliable claims-based summaries. CONCLUSION: Generating patient care summaries based on claims data is feasible and produces valuable information for HCP, provided that the reimbursement process is conducted timely. Integration into a national health telematics platform will facilitate access to the summaries. Evaluation of algorithm and prototype system is underway to prove the benefit in clinical practice. PMID- 17108536 TI - On-the-fly form generation and on-line metadata configuration--a clinical data management Web infrastructure in Java. AB - In this paper we describe the approach to build a web-based clinical data management infrastructure on top of an entity-attribute-value (EAV) database which provides for flexible definition and extension of clinical data sets as well as efficient data handling and high performance query execution. A "mixed" EAV implementation provides a flexible and configurable data repository and at the same time utilizes the performance advantages of conventional database tables for rarely changing data structures. A dynamically configurable data dictionary contains further information for data validation. The online user interface can also be assembled dynamically. A data transfer object which encapsulates data together with all required metadata is populated by the backend and directly used to dynamically render frontend forms and handle incoming data. The "mixed" EAV model enables flexible definition and modification of clinical data sets while reducing performance drawbacks of pure EAV implementations to a minimum. The system currently is in use in an electronic patient record with focus on flexibility and a quality management application (www.healthgate.at) with high performance requirements. PMID- 17108537 TI - Log files analysis to assess the use and workload of a dynamic web server dedicated to end-stage renal disease. AB - A Multi-Source Information System (MSIS), has been designed for the Renal Epidemiology and Information Network (REIN) dedicated to End-Stage Renal Disease (ESRD). MSIS aims at providing reliable follow-up data for ESRD patients. It is based on an n-tier architecture, made out of a universal client, a dynamic Web server connected to a production database and to a data warehouse. MSIS is operational since 2002 and progressively deployed in 9 regions in France. It includes 16,677 patients. We show that the analysis of MSIS web log files allows evaluating the use of the system and the workload in a public-health perspective. PMID- 17108538 TI - Concept representation in health informatics for enabling intelligent architectures. AB - Semantically interoperable health information systems have to be based on shared knowledge and harmonised terminologies/ontologies. Therefore, knowledge representation regarding domain concepts, terms, and relationships used must be harmonised. Starting with the Generic Component Model, different approaches for representing concepts in the healthcare area are discussed, demonstrating common principles and transformation ways. PMID- 17108539 TI - Integrating radiology information systems with healthcare delivery environments using DICOM and HL7 standards. AB - Integration based on open standards, in order to achieve communication and information interoperability, is one of the key aspects of modern health care information systems. Interoperability presents data and communication layer interchange. In this context we identified the HL7 standard as the world's leading medical Information and communication technology (ICT) standard for the business layer in healthcare information systems and we tried to explore the ability to exchange clinical documents with minimal integrated healthcare information systems (IHCIS) change. We explored HL7 Clinical Document Architecture (CDA) abilities to achieve radiology information system integration (DICOM) to IHCIS (HL7). We introduced the use of WADO service interconnection to IHCIS and finally CDA rendering in widely used Internet explorers. PMID- 17108540 TI - The fate of clinical department systems at the dawn of hospital-wide electronic health records in a Norwegian university hospital. AB - OBJECTIVE: a) To document the presence and use of clinical department systems (CDS) in a university hospital that implemented a hospital-wide electronic health record (EHR) in 1999 and b) To compare clinical use of the CDS with that of the EHR. METHOD: Identification of CDSs in use by contacting leaders and senior physicians at clinical departments at the hospital. Identification of key properties of each CDS by interviewing users. RESULTS: We identified a total of 60 CDSs, of which 53 fell in one of four categories; Journal or documentation system tailored to a department or medical specialty (19 systems), Software bundled with electronic medical equipment (control/storage/presentation) (14 systems), Logistics/administration/planning/appointments (10 systems) and Database for medical research (10 systems). Many CDSs were described to outperform the EHR system with regard to ability to provide better patient overview and better support for registering patient data. CDSs are not integrated with the EHR and thus contain islands of data. CONCLUSION: CDSs continue to fill important roles and there is no tendency towards that the hospital-wide EHR makes CDSs obsolete. PMID- 17108541 TI - Software development for the estimation of the mean DRGs related treatment cost. AB - The purpose of the present study was the development and the initial implementation of a Medical Procedure Resources-Allocation and Cost-Capturing Software for the Estimation of the Mean DRGs associated Treatment Cost. The system provides means for the acquisition of health-care cost related data, based upon the actual Greek conditions, and includes Medical Equipment, Reagents, Consumables and Disposable Materials, Drugs, Man Power, Building Leasing and Infrastructure Maintenance expenditures, as the most important cost-components. The developed system was tested in the Operating Rooms (OR) and the Intensive Care Units (ICU), of a representative group of Hospitals. The obtained data have confirmed the reliability of the system for drafting DRGs in Greece; however, there is a discrepancy, between the high actual mean costs in the OR and the ICU, and the relatively low Health Insurance remuneration of the Hospitals. PMID- 17108542 TI - Apply creative thinking of decision support in electrical nursing record. AB - The nursing process consists of five interrelated steps: assessment, diagnosis, planning, intervention, and evaluation. In the nursing process, the nurse collects a great deal of data and information. The amount of data and information may exceed the amount the nurse can process efficiently and correctly. Thus, the nurse needs assistance to become proficient in the planning of nursing care, due to the difficulty of simultaneously processing a large set of information. Computer systems are viewed as tools to expand the capabilities of the nurse's mind. Using computer technology to support clinicians' decision making may provide high-quality, patient-centered, and efficient healthcare. Although some existing nursing information systems aid in the nursing process, they only provide the most fundamental decision support--i.e., standard care plans associated with common nursing diagnoses. Such a computerized decision support system helps the nurse develop a care plan step-by-step. But it does not assist the nurse in the decision-making process. The decision process about how to generate nursing diagnoses from data and how to individualize the care plans still reminds of the nurse. The purpose of this study is to develop a pilot structure in electronic nursing record system integrated with international nursing standard for improving the proficiency and accuracy of plan of care in clinical pathway process. The proposed pilot systems not only assist both student nurses and nurses who are novice in nursing practice, but also experts who need to work in a practice area which they are not familiar with. PMID- 17108543 TI - Benefits and weaknesses of health cards used in health information systems. AB - The acceptance-based success of modern health information systems and health networks highly depends on the respective involvement of all relevant partners into the communication and co-operation processes characterising the medical and administrative workflow. Personal information stored in a networking environment guarantee for fast access fulfilling advanced shared care requirements whereas security token like smart cards stand for identification purposes, data protection, privacy protection, access rights, and limited person-based information storage, e.g., for emergency procedures. Linking these means of information provision allows for making use of the benefits of the different technologies without ignoring their existing weaknesses. The presented paper intends to summarise the respective categories of benefits and weaknesses allowing the reader to implement cards in health information systems as well as with the related aspects of awareness, confidence, and acceptance. Concluding this analysis, the preferred way to deal with the challenges of modern healthcare and welfare requirements shall be a well-balanced combination of cards and networks. PMID- 17108544 TI - A socio-technical study of an ubiquitous CPOE-system in local use. AB - A socio-technical approach was used to study the qualitative effects of deploying a medication-CPOE (Computer-based Provider Order Entry System) at two internal medical wards on a hospital in Denmark. Our results show spatial and temporal transformations of core acts in medication work, transformation in competencies, and less time spent at the bedside for nurses and doctors, as a system- constructed for ubiquitous drug-order entries and handling--was implemented for local use. This study throws light on problems of patient continuity, patient related and IT-system-related error-handling and time spend on core activities- when ubiquitous IT is used locally in a real physical setting with specific traditions of doing medication acts. The paper argues for the project organization to support the local renegotiation of time and place of programs of (work-)action and of collaboration with 'ubiquitous' CPOE-systems, as well as set up feed-back for maturation of software for future clinical use. PMID- 17108545 TI - HIS-monitor: quality of information processing in hospitals. AB - Systematic monitoring of HIS quality is an important task; however, this task is often seen to be insufficiently supported. To support systematic HIS monitoring, we developed the HIS-Monitor questionnaire, focusing on how a hospital information system (HIS) does efficiently support clinical and administrative tasks. HIS-Monitor was applied in a feasibility study with 102 nursing participants. Results point to strengths and weaknesses of information processing in the participating departments. Based on the experiences of the feasibility study, HIS-Monitor is now further being optimized. PMID- 17108546 TI - Development of methods for usability evaluations of EHR systems. AB - Developing electronic health record (EHR) systems in Denmark is an on going, iterative process, where also a maturation process for clinical use should be considered. Convincing methodology for collecting and incorporating in the soft- and hardware knowledge and robustness for the clinical environments is not on hand. A way to involve the clinicians in the development process is conducting usability evaluations. The complexity of the clinical use of the systems is difficult to transmit to a usability laboratory, and due to ethical issues a traditional field study can be impossible to carry out. The aim of this study has been to investigate how it is possible to identify usability problems in an EHR system by combining methods from laboratory tests and field studies. The methods selected for the test design are: the think aloud method, video and screen recording, debriefing, a scenario based on an authentic patient record, and testing on the normal production system. The reliability and validity of the results is increased due to the application of method- and data-triangulation. The results of the usability evaluation include problems in the categories: system response time, GUI-design, functionality, procedures, and error messages. The problems were classified as cosmetic, severe, or critical according to a rating scale. The experience with each method is discussed. It is concluded that combining methods from laboratory test and field study makes it possible to identify usability problems. There are indications that some of the usability problems only occurred due to the establishment of an authentic scenario. PMID- 17108547 TI - Implementing communication systems in the community health services. The health care workers experiences. AB - Reengineering of the workplace through Information Technology is an important strategic issue for today's community health care. The computer-based patient record (CPR) is one technology that has the potential to profoundly modify the work routines of the care unit. This study investigates a CPR project, Gerica aimed at allowing the health care workers in the community health care to work in a completely electronic environment. The focus of our analysis was the use of Gerica, and the health care workers interpretations of it. The rationale behind the introduction of this technology was based on its alleged capability to both enhance quality of care and control costs. This is done by better managing the flow of information within the organization. Theory of structuration is used as the conceptual vehicle to aid in widening the search to the socially constructured nature of these meaning: how people constructed their conceptions in their work setting. The present study analyzed the implementation of CPR conducted in the community health services in Trondheim, Norway. Interviews with Gerica users demonstrate that individual interpretations vary considerably, also between users of the same application. User-resistance was not the problem. This project was a good opportunity to understand better the intricate complexity of introducing technology in professional work where the usefulness of information is short lived and where it is difficult to predetermine the relevancy of information. Profound misconceptions in achieving a tighter fit (synchronization) between care processes and information processes were the main problems. PMID- 17108548 TI - Measurement of the clinical usability of a configurable EHR. AB - The objective of the project was to measure the clinical usability of an EHR configured by use of participatory design with clinicians from a neurological stroke unit in order to get input to the County's future strategy for incremental implementation of EHR. The content of the EHR was defined during a series of workshops with the clinicians after which the XML configuration files were written and deployed. In parallel with this, the participants from the University identified, prioritised and further specified a number of effects related to the clinical practice to be measured. The effects requested by the clinicians focused on improving their overview and assessment of patients as well as on more efficient coordination in three specific and highly cooperative situations, viz. nursing handover, ward round and patient conference. All three situations were measured before (using paper-based medical records) as well as during the week where the configured EHR completely replaced the paper-based medical record in order to compare a 'before' and 'after' situation. Measurements were focused on the requested effects and acquired using various techniques including questionnaires, interviews, observations, and Task Load Index (TLX) ratings. In total, 15 nursing handovers, 8 ward rounds, and 11 patient conferences involving a total of 35 patients and more than 20 clinicians were included in the measurements. Data from the project has been comparatively analysed by means of the TLX scores. Our results show several significant results, for example, during ward rounds the physicians experienced a significant improvement of TLX. The experiment has proven it possible to configure the content of an EHR that significantly improves the clinician's overview of the patient's current status in different clinical situations during the clinical process, based on the clinician's actual needs. Furthermore, the configuration process gave the County valuable experience concerning the content and management of a participatory design process as well as documentation of utility value that will be incorporated in future EHR projects. PMID- 17108549 TI - Ten years of teledermatology. AB - Using telemedicine health professionals can communicate with each other and with their patients over a distance. Teledermatology, dermatology application of telemedicine, is one of the most often applied telemedicine applications worldwide. Various studies have been performed to evaluate the effectiveness and efficiency of and satisfaction with teledermatology. Up to now no or limited valid scientific evidence has been found that teledermatology is beneficial for any group of users. This study aimed to perceive insight into the evolution of evaluation studies of teledermatology over the past ten years in terms of the telemedicine evaluation framework by Holle and Zahlmann consisting of four continuous phases. We added the phase "post implementation studies" that evaluate teledermatology as a fully integrated service in regular care. Retrieved literature from Medline was reviewed by two reviewers independently in order to include studies and classify them into the five phases. Ninety-nine studies out of 372 found unique references were included and classified into the phases. Most represented phase was phase II with 72 (72%) studies. The number of phase II studies is continuously growing since the introduction of evaluation in teledermatology. There were eight reported RCTs found (two in phase III, six in phase IV). The number of phase III and IV studies is too low to draw conclusions about the trends in their publication and stress the need for more such studies. Phase I and post implementations studies are probably under-represented as they might often not be published separately in scientific journal papers. PMID- 17108550 TI - Integrating anatomical pathology to the healthcare enterprise. AB - For medical decisions, healthcare professionals need that all required information is both correct and easily available. We address the issue of integrating anatomical pathology department to the healthcare enterprise. The pathology workflow from order to report, including specimen process and image acquisition was modeled. Corresponding integration profiles were addressed by expansion of the IHE (Integrating the Healthcare Enterprise) initiative. Implementation using respectively DICOM Structured Report (SR) and DICOM Slide Coordinate Microscopy (SM) was tested. The two main integration profiles- pathology general workflow and pathology image workflow--rely on 13 transactions based on HL7 or DICOM standard. We propose a model of the case in anatomical pathology and of other information entities (orders, image folders and reports) and real-world objects (specimen, tissue samples, slides, etc). Cases representation in XML schemas, based on DICOM specification, allows producing DICOM image files and reports to be stored into a PACS (Picture Archiving and Communication System. PMID- 17108551 TI - Medical Data GRIDs as approach towards secure cross enterprise document sharing (based on IHE XDS). AB - Quality and efficiency of health care services is expected to be improved by the electronic processing and trans-institutional availability of medical data. A prototype architecture based on the IHE-XDS profile is currently being developed. Due to legal and organizational requirements specific adaptations to the IHE-XDS profile have been made. In this work the services of the health@net reference architecture are described in details, which have been developed with focus on compliance to both, the IHE-XDS profile and the legal situation in Austria. We expect to gain knowledge about the development of a shared electronic health record using Medical Data Grids as an Open Source reference implementation and how proprietary Hospital Information systems can be integrated in this environment. PMID- 17108552 TI - A multi-agent approach to the design of an e-health system. AB - E-medicine covers the whole range of medical process and service. Multi-agent approach is suitable for the development of e-medicine systems. In this paper, firstly the requirements of e-medicine are analyzed and taxonomy is proposed for e-medicine systems. Secondly multi-agent approach is introduced for developing e medicine systems, and the design of agents and the design of multi-agent structure are presented for e-medicine systems. Finally a case study is presented on a telemedicine for diabetes to illustrate the development of e-medicine systems. Then, our future work is to implement the proposed system. PMID- 17108553 TI - A site of communication among enterprises for supporting occupational health and safety management system. AB - The occupational health and safety management constitutes a field of increasing interest. Institutions in cooperation with enterprises make synchronized efforts to initiate quality management systems to this field. Computer networks can offer such services via TCP/IP which is a reliable protocol for workflow management between enterprises and institutions. A design of such network is based on several factors in order to achieve defined criteria and connectivity with other networks. The network will be consisted of certain nodes responsible to inform executive persons on Occupational Health and Safety. A web database has been planned for inserting and searching documents, for answering and processing questionnaires. The submission of files to a server and the answers to questionnaires through the web help the experts to make corrections and improvements on their activities. Based on the requirements of enterprises we have constructed a web file server. We submit files in purpose users could retrieve the files which need. The access is limited to authorized users and digital watermarks authenticate and protect digital objects. The Health and Safety Management System follows ISO 18001. The implementation of it, through the web site is an aim. The all application is developed and implemented on a pilot basis for the health services sector. It is all ready installed within a hospital, supporting health and safety management among different departments of the hospital and allowing communication through WEB with other hospitals. PMID- 17108554 TI - A new perspective in the promotion of e-health. AB - This paper proposes a new model that provides assistance in understanding the reasons why individuals would use new ICT (Information and Communication Technologies) to perform a healthcare change in their lifestyle. Achieving a lifestyle change by the use of ICT is a multifold issue that can be broadly addressed by analysing, in parallel two dimensions: the individuals' attitude towards their health condition and their approach and readiness to monitor and change their attitude by the use of ICT. Our work has been to conceive and develop a model that explains the different stages the user is at both in terms of the perception of healthcare and of the use of technology to perform any desired or recommended change. In order to place the user at each dimension a set of questionnaires were designed and implemented. These questionnaires assisted us in understanding what personalised information needs to be provided according to the stage the user is at as well as to other variables (such as age, cultural background, etc). The novelty of this model is that it proposes a general framework that may be applied to the conception, design and evaluation of any e health application. Moreover, it can be applied to different application targets (medical informatics, public health informatics, etc) and to different audiences (healthy individuals, patients, professionals, etc) as it proposes an enhanced user modelling process by taking into account both healthcare behaviour aspects as well as technological issues, which up to this moment, have not been taken into account and may be part of the explanation of e-health uptake failure in the healthcare field. The application of this model promotes the empowerment of the individuals by providing tailored information, as well as guidance, monitoring, through ICT and it will certainly make an impact on health-related behaviour. Besides, it will allow to understand some of the reasons of the success or failure of different e-health platforms. Overall, this model is likely to provide deeper insights into the process of improving e-health so it can meet ongoing individuals' needs and become an increasingly valued part of health care services. PMID- 17108555 TI - E-health approach to link-up actors in the health care system of Austria. AB - "Electronic health services are important" the EU commission stated in the E Health action plan. By these means access to health care can be improved and the quality and effect of the offered medical services can be increased. By introducing the e-card in Austria, an overall link-up of nearly all health service providers of the external sector (e.g. family doctors) was achieved. In 2005 the Austrian E-Health Initiative (EHI) of the Austrian Federal Ministry for Health and Women mapped out a strategy to organise the development of the health system towards an integrated patient-centred. Hereby the electronic health record (EHR) plays a decisive role. The aim of this study is to analyse requirements for a virtual, cross-institutional and patient-centred electronic health record from the point of view of the exemplary main actors (Doctor and Patient), to define conditions, and then to evaluate the thus derived, specific concept of implementation. Aside from the two main actors regarding medical acts, namely the institution treating a patient (e.g. doctor, paramedic or nurse) and the patient receiving treatment, a row of other actors could be identified. Group assessment techniques with representatives of these actors resulted in an overview of required functions of an EHR. As a proof-of-concept an information system architecture conformable to the IHE XDS architecture for cross enterprise document sharing is currently being constructed and evaluated in the course of a pilot-project. If the core architecture fulfils the expectations, then a further extension to other hospitals and resident doctors, and subsequently also to the other actors of the health system, is planned. Since both legal and socio technical requirements are presently not yet entirely met, and since there are also deficits from a methodical viewpoint, a complete implementation and widespread introduction will be a long term goal. PMID- 17108556 TI - Architecture for national e-health infrastructure in Lithuania. AB - Building a patient centered National e-Health system has been a challenging task for the Health Ministry of Lithuania in 2005-2006. The first and the main task is to design and to build the infrastructure of the National e-health system. The paper presents the architecture of the system, which aims: (a) to integrate already designed and implemented information systems, (b) to communicate with other national information systems, (c) to provide services (basic ones at the initial stage) for users and (d) to support further e-health systems development in long-term perspective. The system design is based on the HISA approach. The core of the system consists of on-line data storage (including classifiers and registries), of the automation and integration layers for health care processes, of a security layer, a system control layer and a client layer. After having defined the concepts of processes and activities, of the security, of the registries and classifiers within the system and the Electronic Health Records System, then follows the implementation stage. Until the end of 2006 it will include the core of the national e-Health infrastructure and the part of the information system providing basic services for the pilot Lithuanian healthcare institutions. PMID- 17108557 TI - A national EHR strategy preparedness characterisation model and its application in the South-East European region. AB - This paper is concerned with modelling national approaches towards electronic health record systems (NEHRS) development. A model framework is stepwise produced, that allows for the characterisation of the preparedness and the readiness of a country to develop an NEHRS. Secondary data of published reports are considered for the creation of the model. Such sources are identified to mostly originate from within a sample of five developed countries. Factors arising from these sources are identified, coded and scaled, so as to allow for a quantitative application of the model. Instantiation of the latter for the case of the five developed countries is contrasted with the set of countries from South East Europe (SEE). The likely importance and validity of this modelling approach is discussed, using the Delphi method. PMID- 17108558 TI - Simulation based cost-benefit analysis of a telemedical system for closed-loop insulin pump therapy of diabetes. AB - BACKGROUND: INCA (Intelligent Control Assistant for Diabetes) is a project funded by the EU with the objective to improve diabetes therapy by creating a personal closed loop system interacting with telemedical remote control. Cost-benefit analyses of such systems are needed to decide on the introduction of telemedical systems such as the INCA system to routine care. OBJECTIVE: To identify and apply suitable methods for a cost-benefit analysis from the perspective of the payor for health services (i.e. a health insurance company). METHODS: For the cost analysis MOSAIK-M was used, a method and tool that supports health information systems analysis and design. Two MOSAIK-M models were created during the INCA project. Both, the "As is"-model of conventional insulin pump based diabetes care, and the "To be"-model of the INCA system were parameterised with cost values. With both models a period of one year was simulated to determine the yearly costs of diabetes management and treatment for a patient who does not suffer from diabetes related complications yet. The HbA1c-value was chosen as effectiveness parameter for diabetes therapy. To determine the probability of developing complications and their probable duration the Archimedes-Model was used. It was parameterised with selected HbA1c-values anticipating the effect of INCA. The simulation results in form of years of disease within a 30-years time frame were multiplied with corresponding treatment costs from the KoDiM study. RESULTS: The yearly costs of conventional insulin pump treatment for a 19 year old diabetes type 1 patient with no complications are 5,907 euro (German health care system). Using the INCA system would raise the yearly costs by 7,348 euro. Almost all (98.53%) of the additional costs are generated by the continuous blood glucose measurement device. HbA1c-decreases from 7% (conventional treatment) to 6.5%, 6%, and 5.8 % would produce yearly savings (benefit) concerning the treatment of complications of 100.50 euro, 189.20 euro and 221.82 euro. CONCLUSIONS: The selected approach produces an estimation of a lower bound for cost savings. Further work is needed to improve the approximation and to include indirect and intangible costs. The INCA approach would be cost efficient from the chosen perspective, only if the costs of system operation were notably lowered. PMID- 17108559 TI - E-DIMEM: an economic model to estimate the costs of e-disease management. AB - In order to estimate the cost of generic clinical guidelines based on telemedicine services for chronic patients, an economic model (E-DIMEM) has been defined. This model is designed to help health suppliers and managers to plan for the future. It is essentially a workflow composed by diagnosis and therapeutic activities. The cost of each workflow activity is related to healthcare providers (activity agents), drugs, instruments, telemedicine services, and type of specialized healthcare centers used (hospital, clinics, etc.). This model performs multidimensional analysis. A web service based on the E-DIMEM has been implemented. PMID- 17108560 TI - A model for a regional health information network sharing clinical information between professionals in Britanny. AB - The purpose of this paper is to present a generic model of information system supporting healthcare networks for health professionals in Brittany. This model is aimed to develop cooperation between hospital professionals, primary care and practitioners whatever the specific pathology, by providing tools for exchanging and sharing medical-related data. The model is designed based on the heterogeneity factors revealed by a prospective survey. It includes secured exchange of nominative medical data, all other information and activities being accessible by a Web portal. Other associated tools are a synchronous collaborative platform, and a e-learning module. The first implementation, currently in use by the professionals, is presented for the existing neurology healthcare networks. PMID- 17108561 TI - Teleconsultations as a step towards hospital interoperability. AB - The paper presents an interregional telemedicine project between hospitals from two countries that encourage good economic cooperation. It uses modern technology and diverse human resources that contribute to a better accomplishment of the goals of the project and insures continuity of the services after the ending of the project. It instantiates the vision of e-Europe regarding access to healthcare services and the increasing mobility of patients. PMID- 17108562 TI - Health information exchange on a regional level: dream or reality? AB - Cooperation between healthcare institutions on the subject of Information Technology in Healthcare is an important main objective in the region Delft Westland Oostland/Nieuwe Waterweg Noord, in the mid-western part of the Netherlands. This is a densely populated area with approximately 1 million inhabitants. In 2004 a project to uniquely identify patients was initiated. The aim was to build a Regional Referral Index, containing only general patient data, which knows where patient information resides, serving as the fundament of a regional Virtual Electronic Patient Record. Initially the Regional Referral Index was filled with the patient files from two hospitals in the region. A connection was made between the Regional Referral Index and the Hospital Information Systems. Since then, the Regional Referral Index has been updated automatically. The patient files have been tested and matched in order to obtain a 'clean' patient index. Since December 2005 the Regional Referral Index is operational and ready for further developments. Precautions are taken to guarantee patient privacy and security of data. Only authorized users will be given access to patient data. Administration and maintenance of this Regional Referral Index is arranged by establishing a Patient Information Point. Currently, patient files of the other care providers in the region are being added and pilot projects to accomplish access to medical patient information are being realised. The regional activities run parallel and in line with national initiatives, aiming to link up with these initiatives over time. PMID- 17108563 TI - Enabling protocol-based medical critiquing. AB - This paper investigates the combination of expert critiquing systems and formal medical protocols. Medical protocols might serve as a suitable basis for an expert critiquing system because of the ongoing acceptance of medical protocols and the rise of both evidence-based practice and evidence-based protocols. A prerequisite for a critiquing system based on medical protocols is the ability to match the actions a physician performs in practice to actions prescribed by a protocol. Previous research has shown that this is quite difficult, due to the fact that computerized systems are unable to handle deviations from a protocol, which are common in the medical domain. Our solution to this problem is based on extracting the intention underlying a physician's action and uses the intention as the basis for matching performed actions to prescribed actions. We propose an algorithm for the intention-based matching process and we evaluate the matching algorithm on 12 cases of hyperbilirubinemia in healthy term newborns. PMID- 17108564 TI - Management of data quality--development of a computer-mediated guideline. AB - Appropriate data quality is a crucial issue in the use of electronically available health data. As source data verification (SDV) and feedback are two standard procedures for measuring and improving data quality it would be worthwhile to adapt these procedures to a current level of quality in order to reduce costs in data management. This project aims to develop a guideline for the management of data quality with special emphasis on this adaptation against the backdrop of research networks in Germany, which operate registers and conduct epidemiological studies. The first step in guideline development was a thorough literature review. The literature offers many measurements as candidates for quality indicators, however, systematic assessments and concepts of SDV and feedback are missing. We assigned possible quality indicators to the levels plausibility, organization, and trueness. Each indicator must be operationally defined to allow automatical calculation. The SDV sample size calculation leads to lower numbers for sites providing data of good quality and larger numbers for sites with poor data quality. The guideline's implementation in a software tool combines two cycles, one for the adaptation of recommendations to a given study/register, the other for the improvement of data quality in a PDCA-like approach. The recommendations will address needs common to medical documentation in daily health care, clinical, epidemiological, and observational studies as well as in surveillance data bases and registers. Further work will have to supplement other aspects of data management. PMID- 17108565 TI - Design of a decision support system for chronic diseases coupling generic therapeutic algorithms with guideline-based specific rules. AB - Clinical Decision Support Systems (DSS) help improve health care quality. They usually incorporate an Execution Engine (EE), defined for each disease. We have designed, and present here, a generic execution engine, coupled with guideline based disease specific rules stored in knowledge base (KB) as part of the prescription-critiquing mode of the ASTI project. This system was designed using two national guidelines for type 2 diabetes and hypertension. It takes into account the patient's clinical data, the tolerance and effectiveness of previous and current treatments and the physician's prescription made at the time. The functioning of the system has been speeded up and its maintenance made easier by indexing the KB rules according to the type of treatment they are linked to (e.g. monotherapy, etc.) and by classifying them into four categories. The EE's design formalizes generic therapeutic algorithms, leading to treatment options for cases of bad tolerance or insufficient effectiveness of the current treatment. Its applicability to other diseases was shown by applying it to dyslipidemia. PMID- 17108566 TI - Automated test selection in decision-support systems: a case study in oncology. AB - Decision-support systems in medicine should be equipped with a facility that provides patient-tailored information about which test had best be performed in which phase of the patient's management. A decision-support system with a good test-selection facility may result in ordering fewer tests, decreasing financial costs, improving a patient's quality of life, and in an improvement of medical care in general. In close cooperation with two experts in oncology, we designed such a facility for a decision-support system for the staging of cancer of the oesophagus. The facility selects tests based upon a patient's health status and closely matches current routines. We feel that by extending our decision-support system with the facility, it provides further support for a patient's management and will be more interesting for use in daily medical practice. In this paper, we describe the test-selection facility that we designed for our decision-support system in oncology and present some initial results. PMID- 17108567 TI - Avoiding literature overload in the medical domain. AB - The retrieval of patient-related literature is hampered by the large size of medical literature. Various computer systems have been developed to assist physicians during information retrieval. However, in general, physicians lack the time and skills required to employ such systems effectively. Our goal is to investigate to what extent a physician can be provided with patient-related literature without spending extra time and without acquiring additional skills. In previous research we developed a method to formulate a physician's patient related information needs automatically, without requiring any interaction between the physician and the system. The formulated information needs can be used as a starting point for literature retrieval. As a result we found that the number of information needs formulated per physician was quite high and had to be reduced to avoid a literature overload. In this paper we present four types of knowledge that may be used to accomplish a reduction in the number of information needs. The usefulness of each of these knowledge types depends heavily on the specific cause underlying the multitude of information needs. To determine the nature of the cause, we performed an experimental analysis. The results of the analysis led us to conclude that the knowledge types can be ordered according to their appropriateness as follows: (1) knowledge concerning temporal aspects, (2) knowledge concerning a physician's specialism, (3) domain knowledge, and (4) a user model. Further research has to be performed, in particular on precisely assessing the performance of each type of knowledge within our domain. PMID- 17108568 TI - A language classifier that automatically divides medical documents for experts and health care consumers. AB - We propose a pipelined system for the automatic classification of medical documents according to their language (English, Spanish and German) and their target user group (medical experts vs. health care consumers). We use a simple n gram based categorization model and present experimental results for both classification tasks. We also demonstrate how this methodology can be integrated into a health care document retrieval system. PMID- 17108569 TI - With good Intentions. AB - The use of intentions in computer-based guidelines may help to make them more flexible, easier to adapt to local standards, easier to evaluate and to improve. We see possibilities of using intentions in several areas: as a generator of actions which could be performed and then compared with the decision of the physician, as a method for pruning proposed actions based on current and previous patient data. In this contribution we present our approach to intention-based guidelines. PMID- 17108570 TI - A method for specification of structured clinical content in electronic health records. AB - The Copenhagen County is using clinical guidelines in the electronic health record development to provide documentation support, process support and decision support for the healthcare professionals. The electronic health record development is based on three main components: The first component is a national information model. The second component is a common classification system (SNOMED). The third key component is the so-called "clinical content". This paper describes the structured "clinical content", how it is linked to the clinical process, and how it is used to create clinical guidelines in the form of standard care plans. The Copenhagen County and MEDIQ has developed a methodology for identifying and specifying structured "clinical content" to be used in electronic health records. The method combines analyses of national clinical guidelines with local experience and practices and it heavily involves healthcare professionals. The method includes four main steps: Analyses of background material, analyses of clinical process-flow, mapping to standards (the national information model and the common classification system), and specification of the structured clinical content itself. Three secondary steps may be added to specify the clinical content in more detail: Workflow analyses, analyses of quality indicators, and decision analyses. This paper reports the experiences using the method and stresses the demand for a common exchange format and IT-tools for documenting clinical content in a formalised way. PMID- 17108571 TI - Using Treemaps to represent medical data. AB - Confronted with the inadequacy of traditional charts, we tested the contribution of Treemaps to the representation of medical data. Treemap charts allow description of large hierarchical collections of quantitative data, on a synthetic way. Treemaps were implemented using PHP5, and were tested in the field of DRG-mining and other medical informations. From now on, this implementation is used in an interactive web-based request tool, and could be used to design interactive piloting tools. PMID- 17108572 TI - Evidence in pharmacovigilance: extracting adverse drug reactions articles from MEDLINE to link them to case databases. AB - Literature, specifically MEDLINE, is among the main sources of information used to detect whether a drug may be responsible for Adverse Drug Reactions cases. The aim of our work is to automate the search of publications that correspond to a given Adverse Drug Reactions case: (i) by defining a general pattern for the queries used to search MEDLINE and (ii) by determining the threshold number of publications capable to confirm or infirm the Adverse Drug Reaction. We applied our algorithm to a set of 620 cases from a French pharmacovigilance database. We obtained a precision of 93%, recall 70%. We determined a threshold of 3 publications to confirm an Adverse Drug Reaction case. PMID- 17108573 TI - Towards the use of ontologies for activity-based costing in healthcare organizations. AB - Activity-based costing is a methodology which provides a basis for healthcare cost optimization. A robust formal representation of such activities is needed in order to be able to draw inferences as to the costs involved in a reliable manner. An introduction to the basic ontological distinctions involved in such a representation and to the complications one faces in carrying out the task is presented in this paper. PMID- 17108574 TI - From bibliometric analysis to research policy: the use of SIGAPS in Lille University Hospital. AB - In French hospitals, the progressive setting up of the new rating systems has obliged the university hospitals to justify a certain amount of activities such as research, training or moreover recourse, which are specific missions of the university hospitals. In order to justify research activities, the Lille University Hospital has developed for now three years SIGAPS, a full-web bibliometric software which census and analyse, the scientific publications referenced in the Medline database. After data downloading, each article is classified on a 6 levels "quality scale derived from the impact factors. The system then performs, for a researcher or a team, a report allowing a quantitative and qualitative analysis. Started in Lille in November 2004, the inventory and analysis of data is now ending. For the period 2001-2004, 2814 articles have been published in 700 different journals. The total number of articles increased from 688 in 2001 to 757 in 2004. The mean impact factor was equal to 2.26 and 15.5 % of articles were classified as A, 20.9% as B. Those results confirm the high level of research of the University Hospital of Lille, in agreement with two other national studies which ranks our establishment at the 6th position for medical research activities among the French University Hospitals. Currently a similar evaluation has now began in the 9 other university hospital which have subscribed to the SIGAPS project. We works currently on new indicators as patents, thesis or conferences, or access to other databases as Sciencedirect or Scopus via the RIS format. The next step in the project is the implementation of a meta-base which will federate the information provided by each SIGAPS system. This meta-base will then allow us to perform comparisons between different hospitals, determine the national "sites of excellence" and create some clinical and research networks. PMID- 17108575 TI - Dealing with an information overload of health science data: structured utilisation of libraries, distributed knowledge in databases and Web content. AB - The organizational structures of web contents and electronic information resources must adapt to the demands of a growing volume of information and user requirements. Otherwise the information society will be threatened by disinformation. The biomedical sciences are especially vulnerable in this regard, since they are strongly oriented toward text-based knowledge sources. Here sustainable improvement can only be achieved by using a comprehensive, integrated approach that not only includes data management but also specifically incorporates the editorial processes, including structuring information sources and publication. The technical resources needed to effectively master these tasks are already available in the form of the data standards and tools of the Semantic Web. They include Rich Site Summaries (RSS), which have become an established means of distributing and syndicating conventional news messages and blogs. They can also provide access to the contents of the previously mentioned information sources, which are conventionally classified as 'deep web' content. PMID- 17108576 TI - A Markov model to describe daily changes in organ failure for patients at the ICU. AB - As the support and stabilization of organ function is a major goal of treatment in the Intensive Care Unit (ICU), changes in the function of organ systems are an important indicator of the progression of the disease and recovery. This paper presents how to construct a model that describes changes in organ failure of ICU patients on a day-to-day basis. The model is based on the daily Sequential Organ Failure Assessment (SOFA) scores for six organ systems and predicts, for each of these organ systems, whether failure or recovery is due on the next day, using six logistic regression equations. The joint set of equations, extended with equations for predicting ICU discharge and death, constitutes a firstorder multivariate Markov model. We applied the procedure on a dataset and found that most types of organ failure are highly persistent. PMID- 17108577 TI - Online guideline assist in intensive care medicine--is the login-authentication a sufficient trigger for reminders? AB - INTRODUCTION: Rising cost pressure due to the implementation of the DRG-System and quality assurance lead to an increased use of therapy standards and standard operating procedures (SOPs) in intensive care medicine. The intention of the German Scientific Society supported project "OLGA" (Online Guideline Assist) is to develop a prototype of a knowledge based system supporting physicians of an intensive care unit in recognizing the indication for and selecting a specific guideline or SOP. While the response of the prototype on user entries can be displayed as a signal on the used workstation itself, the location and time for a reminder of scheduled or missed procedures or reactions to imported information is a difficult issue. One possible approach to this task is the display of non acknowledged reminders or recommendations while logging on to a system. The objective of this study is to analyse user behaviour of the physicians working on the surgical intensive care unit to decide whether the login authentication is a sufficient trigger for clinical reminding. METHODS: The surgical intensive care unit examined in this study comprises 14 beds. Medical care is provided by physicians working in shifts 24 hours a day, 7 days a week, with two anaesthetists at a time and an additional senior consultant during daytime. The entire documentation (examinations, medication, orders, care) is performed using the patient data management system ICUData. The authentication process of the physicians was logged and analysed. RESULTS: Throughout the observation period from December 13th 2005 to January 11th 2006 3563 physician logins were counted in total. The mean span between logins was in 11.3 minutes (SD 14.4), the median 7 minutes. The 75% centile was 14 minutes, the 95% centile 38 min. Intervals greater than 60 minutes occurred in 75%, and greater than 90 minutes in 25% of the days. DISCUSSION: It seems reasonable that reminders sent during authentication are able to enforce workflow compliance. It is possible to send notifications caused by external events to the physician depending on the importance of the event. Serious events with high urgency should be reliably passed using wireless pager or handheld technology. It seems that after the implementation of the prototype guideline assist further investigation is needed to monitor changes in authentication behaviour and reactions to the guideline advisory. This is also required to investigate the influence of unit's size, medical specialty and actual ward workload. PMID- 17108578 TI - Functional data analysis for gait curves study in Parkinson's disease. AB - In Parkinson's disease, precise analysis of gait disorders remains essential for the diagnostic or the evaluation of treatments. During a gait analysis session, a series of successive dynamic gait trials are recorded and data involves a set of continuous curves for each patient. An important aspect of such data is the infinite dimension of the space data belong. Therefore, classical multivariate statistical analysis are inadequate. Recent methods known as functional data analysis allow to deal with this kind of data. In this paper, we present a functional data analysis approach for solving two problems encountered in clinical practice: (1) for a given patient, assessing the reliability of the gait curves corresponding to the different trials (2) performing intra individual curves comparisons for assessing the effect of a therapy. In a first step, each discretized curve was interpolated using cubic B-splines bases in order to ensure the continuous character of data. A cluster analysis was performed on the smoothed curves to assess the reliability and to identify a subset of representative curves for a given patient. Intra individual curves comparisons were carried out in the following way: (1) functional principal component analysis was performed to describe the temporal structure of data and to derive a finite number of reliable principal components. (2) These principal components were used in a linear discriminant analysis to point out the differences between the curves. This procedure was applied to compare the gait curves of 12 parkinsonian patients under 4 therapeutic conditions. This study allowed us to develop objective criteria for measuring the improvements in a subject's gait and comparing the effect of different treatments. The methods presented in this paper could be used in other medical domains when data consist in continuous curves. PMID- 17108579 TI - A framework for cohesive healthcare coalition formation. AB - The mobilisation of cohesive and effective groups of healthcare human resource is important in ensuring the success of healthcare organisations. However, forming the right team or coalition in healthcare organisations is not always straightforward due to various human factors. Traditional coalition formation approaches have been perceived as 'materialistic' or focusing too much on competency or pay-off. Therefore, to put prominence on the human aspects of working together, we present a cohesiveness-focused healthcare coalition formation methodology and framework that explores the possibilities of social networks, i.e. the relationship between various healthcare human resources, and adaptive resonance theory. PMID- 17108580 TI - An approach for generating fuzzy rules from decision trees. AB - Identifying high-risk breast cancer patients is vital both for clinicians and for patients. Some variables for identifying these patients such as tumor size are good candidates for fuzzification. In this study, Decision Tree Induction (DTI) has been applied to 3949 female breast cancer patients and crisp If-Then rules has been acquired from the resulting tree. After assigning membership functions for each variable in the crisp rules, they were converted into fuzzy rules and a mathematical model was constructed. One hundred randomly selected cases were examined by this model and compared with crisp rules predictions. The outcomes were examined by the area under the ROC curve (AUC). No significant difference was noticed between these two approaches for prediction of recurrence of breast cancer. By soft discretization of variables according to resulting rules from DTI, a predictive model, which is both more robust to noise and more comprehensible for clinicians, can be built. PMID- 17108581 TI - Computational representation of Alzheimer's disease evolution applied to a cooking activity. AB - This article presents a computational model and a simulation of the decrease of activities of daily living performances due to Alzheimer's disease. The disease evolution is simulated thanks to the cognitive architecture ACT-R. Activities are represented according to the retrieval of semantic units in declarative memory and the trigger of rules in procedural memory. The simulation of Alzheimer's disease decrease is simulated thanks to the variation of subsymbolic parameters. The model is applied to a cooking activity. Simulation of 100 hundred subjects shows results similar to those realised in a standardized assessment with human subjects. PMID- 17108582 TI - Strategies for health information retrieval. AB - BACKGROUND: The amount of health data accessible on the Web is increasing and Internet has become a major source of health information. Many tools and search engines are available but medical information retrieval remains difficult for both the health professional and the patients. OBJECTIVE: In this paper we describe heuristics that aim at matching as much as possible queries with the content of the documents in the context of the CISMeF catalogue (Catalogue and Index of Health Resources in French) and its Doc'CISMeF search tool. The queries are represented by terms and the content of the documents is indexed by a terminology based on the MeSH thesaurus. RESULTS: Several operations are performed to match the terms of the terminology: natural language processing techniques on multi-words queries, phonemisation, spelling correction, plain text search with adjacency etc.. Each one is tested to evaluate its contribution in matching the terminology and the indexed documents. CONCLUSION: The implemented heuristics contribute significantly with good results in maximising as much as possible the recall of the Doc'CISMeF search tool. PMID- 17108583 TI - A method of cross-lingual consumer health information retrieval. AB - OBJECTIVES: This paper presents a method of cross-language information retrieval aiming to make medical information available to patients in French and English, regardless of the query language they wish to use. METHODS: We describe the two MeSH-related terminologies used in this work. We show that the French patient synonyms included in CISMeF can be automatically mapped to the English consumer oriented health topics used in MEDLINEplus, via the MeSH thesaurus. The links between French and English patient terms thus inferred can subsequently be exploited to automatically translate patient queries. RESULTS: 129 MEDLINEplus topics have been mapped to 142 CISMeF patient synonyms. Contextual links for cross-language retrieval have been added to the patient dedicated French information Gateway CISMeF. CONCLUSION: we have presented an efficient method for cross-lingual patient information retrieval in French and English, which may also be applied to other language pairs, subject to the availability of patient terminologies and of the MeSH thesaurus in these languages. PMID- 17108584 TI - Applying an artificial neural network to predict osteoporosis in the elderly. AB - Osteoporosis is an essential index of health and economics in every country. Recognizing asymptomatic elderly population with high risks of osteoporosis remains a difficult challenge. For this purpose, we developed and validated an artificial neural network (ANN) to identify the osteoporotic subjects in the elderly. The study population consisted of 1403 elderly adults (mean age 63.50 +/ 0.24 years ranged from 50 to 91 years old, 157 male and 1246 female) randomly selected into 3 sets, 703 participants in training set, 350 participants in selection set, and the remaining 350 participants in test set. The input variables included demographic characteristics, anthropometric measurements, and clinical data. The outcome variable was dichotomous, either non-osteoporotic (T score of greater than -2.5) or osteoporotic (T-score of -2.5 or less) groups classified by the measurement from dual energy X-ray absorptiometry. ANN was constructed with data from training and selection sets and validated in test set whose outcome variable was unknown to the network. The performance of ANN was evaluated by discrimination and calibration simultaneously. After training processes, the final best ANN was a multilayer perceptron network which determined seven input variables (gender, age, weight, height, body mass index, postmenopausal status, and coffee consumption) as significant features. The discriminatory power of ANN for test set was excellent (area under receiver operating characteristics curve = 0.82 +/- 0.03). ANN also had statistically good fit represented by statistically insignificant Hosmer-Lemeshow statistic (p = 0.24). These results suggested that our final ANN concurrently had good discriminatory power and good-fit calibration. ANN can be used as a promising tool for the elderly to stratify high risk subjects into osteoporotic group. PMID- 17108585 TI - Design and evaluation of a computer reminder system to improve prescribing behaviour of GPs. AB - OBJECTIVE: To evaluate the implementation of a decision support system with reactive computer reminders to improve drug prescribing behaviour. METHODS: A clustered RCT with an incomplete block design was carried out in the south of the Netherlands: 25 GPs received reminders on antibiotics and asthma/COPD prescriptions, 28 GPs received reminders on cholesterol prescriptions. Prescribing guidelines were integrated into the GP information system, which was installed in the GPs practices of the intervention group. When the computer program was in use, a reminder popped up if the GP deviated from the guidelines during prescribing. PRIMARY OUTCOME: prescription according to the guidelines as a percentage of total prescriptions of a specific drug. Furthermore, an evaluation on the user-friendliness of the CRS in the GP's practice was carried out through questionnaires and interviews. RESULTS: Presently analyses are being carried out. Preliminary results indicate that the CRS study supported our expectations. In general, there seems to be a reduction in the numbers of prescriptions according to the advices of the computerised guidelines not to prescribe certain drugs. Final analysis will be performed shortly. In general, the Computer Reminder System was perceived as stable and user friendly. CONCLUSION: We created a stable and user friendly Computer Reminder System which was adjusted to the needs and demands of GPs. Preliminary results regarding the effectiveness of the system seem to indicate that the implementation of a Computer Reminder System with reactive reminders improves drug prescribing behaviour. PMID- 17108586 TI - Clinicians' perceived usefulness of a support system for patient-centered cancer care. AB - We investigated the perceived usefulness by 65 nurses and 12 physicians who had used CHOICE, a support system designed to improve patient-centred symptom management for cancer patients at the point of care. Two questionnaires addressed the following aspects: clinicians' usage patterns; ease of use; system ability to improve care planning, understanding of patients' perspectives, and patient provider communication; attitudes towards patients' involvement in decision making about patient care; and perceived usefulness, defined as a system's ability to enhance work performance. The overall survey response rate was 78%. Clinicians reported that they had used information outputs provided by the CHOICE system on average 50% of the time, but nurses used them significantly more than physicians. The system received high ratings on all aspects of usefulness by both groups, but again, nurses provided consistently higher usefulness ratings than physicians did. There was a strong, significant correlation between patterns of use and perceived usefulness. There were no correlations between perceived usefulness and respondents' age, gender and clinical experience. Results confirm that the CHOICE system can successfully assist nurses and physicians to improve patient care for cancer patients in ongoing practice. PMID- 17108587 TI - Evaluation of a breast cancer computer aided diagnosis system. AB - Mammography is accepted as the most effective method to detect breast cancer. However, interpreting a mammogram is not easy for not experienced radiologists. The aim of computer aided detection techniques in breast cancer is to improve the chance that a malignant region is detected and appropriately evaluated. Breast microcalcifications have been considered as a very useful index of malignancy, which helps in the early detection of breast cancer. A system of computer aided diagnosis has been developed that is based on detailed analysis and evaluation of related features of individual microcalcifications and of formed clusters helping the doctor to make risk estimation for each microcalcification cluster as well as for isolated microcalcifications. This information is considered to be very useful to radiologists, giving them extra input before making their estimation of each case. The aforementioned system has been thoroughly tested using a number of real life cases provided from collaborating doctors. Each case, apart from the mammograms, was accompanied by a biopsy test result, the patient's demographic data and medical history. A total of 200 cases (147 benign and 53 malignant) have been examined and the results are presented as the Receiver Operating Characteristic (ROC) performance and are quantified using the ROC curve. The system is showing high levels of sensitivity identifying correctly all malignant cases. PMID- 17108588 TI - Comparison of the impact of cardiovascular guidelines on a working population. AB - The influence of guidelines is more and more important in the medicine using standardised knowledge as Clinical Practice Guideline (CPG). OBJECTIVES: 1) Determine the impact of different CPGs on a working population 2) Using the Framingham risk calculation as gold standard to check the CPG classifications. METHODS: This work is done in the context of two hypertension management CPGs published in 1997 and 2000, and one dyslipidemia CPG by the French agency HAS. A French regional study sample of 2817 patients was selected. The results of the classification system by CPG were compared to those given by the chosen gold standard: Framingham cardiovascular risk calculation. RESULTS: The HBP CPG concerns more of the quarter of the working population. The mean of the cardiovascular risk calculation for the patients with drug treatment is higher than for the patients with no drug treatment and for those not screened by the CPG. Some patients with a very high calculated risk, are not selected by the screening CPG based on too few or on not adequate variables. CONCLUSIONS: For the two High Blood Pressure Management CPGs, the selected patients are not distributed the same way in the two CPGs. Observed agreement is poor between the two hypertension CPG versions. Screening people for specific health care should be based on risk calculation not on few variables. Feeding back our results to the CPG creator group should reduce the variability of recommendations. PMID- 17108589 TI - IT support for clinical pathways--lessons learned. AB - Clinical pathways are an effective instrument to decrease undesired practice variability and improve clinician performance. IT applications embedded into clinical routine work can help to increase pathway compliance. Successfully implementing such applications requires both a responsive IT infrastructure and a participatory and iterative design process aimed at achieving user acceptance and usability. Experiences from the implementation and iterative improvement of an online surgical pathway at Marburg University Medical Centre have shown that pathway conformance actually could be improved by the use of IT. An analysis of the iterative design process has shown that future pathway projects can benefit from the lessons learned during this project. Based on these lessons a method for developing well adapted interaction mechanisms is presented, which is aimed at improving process alignment. Our goal is to build up a library of tested reusable components to reduce the number of iterations for pathway implementation. PMID- 17108590 TI - Evidence-based practice in primary health care. AB - This study aims to describe primary care professionals' self-reported attitudes towards evidence-based practice (EBP), attention to information sources, perceptions of the barriers to EBP and strategies to improve insight in EBP and patient care. An e-mail invitation with link to an Internet-based survey was sent to Belgian medical doctors (MDs), nurses and paramedics. Under paramedics, we've included emergency medical technicians, firemen and medical volunteers (Red Cross). In general, respondents were supportive towards EBP and agreed that this concept improves patient care, but still, physicians claim that only 50% of their practice is evidence-based and nurses and paramedics spend respectively 59% and 54% of their time to EBP. Doctors depend mostly on clinical guidelines, the Internet and textbooks, while nurses prefer conferences and protocols and paramedics rely on courses and their own judgement. All respondents strongly rely on experimental knowledge gained through interaction with colleagues, although the majority reported that colleagues are often not supportive towards EBP. Lack of time, the overwhelming mass of literature, difficulties with implementation of evidence in to practice are the most common barriers. Nurses show lack of critical appraisal of research results and paramedics have difficulties understanding research and have limited access to computer facilities and their working environment. Communication in group and workshops are very highly valued. Nurses and paramedics are less reluctant towards the opinion of senior colleagues, audits on clinical practice and individual feedback than doctors. EBP generally enjoys a positive attitude at every level of the health care system, but still many obstacles have to be overcome to conquer 'experience-based practice'. The most appropriate method for actual implementation of evidence based practice at all levels of health care is to provide summaries of evidence, easily understandable protocols and web-based databases accessible from the working environment. Students should not only learn the skills related to EBP, but should be able to integrate knowledge effectively in the clinical setting and routine care. Above all, their supervisors themselves need to evolve from 'experience-based' to evidence-based practice'. PMID- 17108591 TI - Process frame instances for integrating strategic, tactical and operational information management in hospitals. AB - An approach to reduce the complexity of information management is to distinguish it into strategic, tactical and operational management with each of the levels using its own software tools. In practice, the management levels are tightly connected and interact closely. The most evident information that has to be interchanged between the management levels is the information about finished, cancelled, running and planned activities or projects, respectively. We claim that the levels of information management should share information about the work done, and propose their integration by means of a network of so-called process frame instances. Process frame instances hold information about the activities of different information management levels in a structured and reusable way. PMID- 17108592 TI - Web services based syndromic surveillance for early warning within French Forces. AB - OBJECTIVES: Syndromic surveillance for early warning in military context needs a robust, scalable, flexible, ubiquitous, and interoperable surveillance system. METHODS: We have designed our surveillance system as a collaborative network of web services on the basis of a skill oriented decomposition of the overall task and a formal model of epidemiological events. RESULTS: The services (integration devices, epidemiologic receivers, information processing devices, GUI clients) are distributed in several locations in France and French Guiana using a secured network. CONCLUSIONS: This system is in operation since several months. It has already early detected two outbreaks before conventional surveillance systems. PMID- 17108593 TI - Modeling economic aspects of hospital information systems to give decision support for strategic information management. AB - Information systems require strategic planning in order to adapt their functionality and quality to the needs of health care organizations. Next to effectivity, cost efficiency in supplying and operating information systems is a particular objective. Hospital information systems with their technical infrastructure, their application systems and the hereby supported business functions can be described with the help of the meta model 3LGM2 and the 3LGM2 tool. The meta model and the 3LGM2 tool are extended by a generic approach to show supply and operation cost for all components of the information system and for the cost calculation between these components. This leads to the fact that all executives in hospitals are enabled to get the cost transparent which were caused by the support of the functions by the information system. The effects of planned extensions and modification of the information system can be analyzed in term of cost. In a prototypical modelling, the information system of a hospital of regular standards has been evaluated in nearly all its components and cost. An evaluation could show that information managers and executives are now delivered relevant cost information for planning, operating and control of information systems. PMID- 17108594 TI - Medical error management and the role of information technology--a new approach to investigating medical handover in acute care settings. AB - While identifying reasons why medical errors occur and constructing models of how to manage them has proved relatively straightforward, implementing and meaningfully evaluating solutions in 'real-world' settings has proven considerably more difficult. From an information systems (IS) perspective, although the promise of technology remains powerful, the continuing high incidence of medical errors suggest that eHealth approaches are struggling to acquire a clear understanding of the complex, dynamic and multi-layered nature of acute care settings and clinical practices, and to respond effectively to address the range of errors that actually occur. Using medical handover as a field-site, this research-in-progress paper presents an adaptation of James Reason's 'Swiss Cheese Model' to conceptualize the complex factors at play in medical errors in terms of human, system and informational elements. This research paper then examines how drawing on this model it is possible to generate and implement a methodological approach that both enhances a holistic understanding of medical error management and illuminates criteria that can be used to meaningfully identify an appropriate role for information technology in medical error mitigation. This research-in-progress paper aims to make a significant contribution to research into medical error management in 'real-world' acute care settings. This research is part of a bigger project that aims to develop, implement and evaluate an information technology artefact as part of an holistic information systems approach to improving medical error management at medical handover. PMID- 17108595 TI - Human factors engineering for clinical applications. AB - This position paper advocates the necessity for promoting a Human Factors Engineering (HFE) approach to new clinical applications in the medical informatics domain. We first describe the negative consequences of NOT using HFE methods for such sensitive applications as medication Computerized Physicians Order Entry systems. We then describe rapidly the HF technology and how it would apply to the healthcare domain. The specific and complex problem of the necessary re-engineering of existing applications is then addressed. We conclude on the mandatory cooperative characteristic of any practical HFE project for designing or re-engineering safety-sensitive clinical applications. PMID- 17108596 TI - Modeling a health telematics network: does the 3LGM2 approach assist in its management and operation ? AB - Health Telematics Networks (HTN) are characterized by a complex setup and interrelations. Using available tools and methods focusing on mainly one aspect, e.g. its functionality or the network infrastructure leads to a restricted view. The objective of this paper is to assess the applicability of the Revised Three layer Graph-based Meta Model (3LGM(2))--developed for modeling hospital information systems--towards health telematics networks. Having identified an approach on how to represent the hospitals effectively in the model the 3LGM(2) proved to support strategic management, day-to-day maintenance and documentation. PMID- 17108597 TI - Agent based simulations in healthcare. AB - Agent Based Simulations (ABS) is a relatively recent computer paradigm. As opposed to "top down" conventional computer simulations, the ABS approach is a "bottom-up" modelling technique where a medium to high number of independent agents is modelled. These agents' interactions sometimes cause unexpected "emergent" system behaviour. ABS is particularly suitable in the social context such as healthcare where a large number of human agents interact and co-operate for common goals. Today ABS in the social context is often used together with the recently introduced network analysis techniques and network visualization tools for modelling and simulating social agents within organisations. At Akdeniz University we are starting a number of projects for applying ABS technology in healthcare. In this paper we present two of the ongoing projects in this field. Firstly we have developed a prototype simulator for the long term monitoring of Chronic Obstructive Pulmonary Disease (COPD) as a major public health problem. We present the COPD simulator, its agents, parameters and working principles. Secondly we want to apply ABS and the network analysis techniques to visualise and explore informal social networks amongst staff at the Akdeniz University Hospital to assess and evaluate properties of the organisation in terms of its ability to innovate and share knowledge. In our applications, we primarily aim to use ABS in a web-based platform to create a virtual environment for discussion, visualising and running what-if scenarios to test out various options for managing healthcare, as well as sharing information and creating a virtual community. PMID- 17108598 TI - Change readiness research. A qualitative study of variations in participation. AB - The Change readiness research method (CRR) has become a well- known method in Denmark to identify issues needed to be discussed on a hospital ward before implementation of a new IT-system and to start a dialogue. A precondition for a constructive dialogue, however, is a high degree of participation. The latest experiences of the CRR method were gained from its use in eight wards in the Danish Gepka project during 2003-4 (The Gepka project was established by The Danish Ministry of the Interior and Health, The National Board of Health, the County Council Society and H:S. Its purpose is to validate the "Basic Structure for The Electronic Health Record" (B-EHR) using prototypes. http://medinfo.dk/epj/proj/gepka/). In the Gepka project the participation varied from 33.3% to 78.9%. The objective of this study is to set out themes by which this variation can be studied. A qualitative explorative research design has been applied, where four instructions from the "Instruction for use" (Instructions for using the CRR method. Can be downloaded the Internet: (http://www.epjobservatoriet.dk/publikationer/forandringsparathed.pdf)) have been studied as themes. The methods used have been telephone interviews and direct observations. The results showed that the seven wards (one was excluded) followed the "Instructions for use" to different degrees. It was found that one instruction, in particular, seems to be especially important to follow to motivate the employees on a ward to participate in the CRR; the management of the ward must be engaged/actively involved in the project, as they are key figures when it comes to motivating the other ward employees. The aim of this study is not to prove a causal relationship between the degree to which the "Instructions for use" are followed and the degree of participation--it is to suggest a qualitative relationship between the two. Neither does this study try to generalize the results, as further research on more wards would be needed to do so. This study does, however, set out themes that can be a useful tool in future CRR projects in order to maximize the degree of participation. In a modified way, these themes can probably be used as a tool in other studies of human-machine interactions. PMID- 17108599 TI - A study of the communication notes for two asynchronous collaborative activities. AB - INTRODUCTION: To build relevant tools for Health Care Professionals, we must study and understand their practices. This paper discusses the way they leave traces in the Patient Record to help asynchronous collaboration, elaborating new documents or adding annotations. METHODS: We compared the results of two studies about the various writing strategies used by the Health Care Professionals to capture knowledge in the Patient Records. The first study deals with the information written by the nurses in a textbook during homecare situations. The second one deals with the annotations left by all the practitioners to complete the documents of the patient record in a hospital ward. RESULTS: We have found some invariants in these two situations. An interpretation model based on four levels: Communication Context, Communication Object, Value of Communication and Value of Cooperation, is proposed in order to describe and to index the characteristics of the Communication Notes. PMID- 17108600 TI - Specific classification of elibrary resources says more about users' preferences. AB - BACKGROUND: Medical Subject Headings (MeSH) are a hierarchical taxonomy of over 42,000 descriptors designed to classify scientific literature; it is hierarchical with generic high order headings and specific low order headings. Over 1,000 resources in the Primary Care Electronic Library (PCEL - www.pcel.info) were classified with MeSH. METHODS: Each of the entries or resources in the primary care digital library was assigned up to five MeSH terms. We compared whether the most generic or specific MeSH term ascribed to each resource best predicted user preferences. RESULTS: over the four month period analysed statistically significant differences were found for resources according to specific key MeSH terms they were classified by. This result was not repeated for generic key MeSH terms. CONCLUSIONS: Analysis of the use of specific MeSH terms reveals user preferences that would have otherwise remained obscured. These preferences are not found if more generic MeSH terms are analysed. PMID- 17108601 TI - Cancer patient flows discovery in DRG databases. AB - In France, cancer care is evolving to the design of regional networks, so as to coordinate expertise, services and resources allocation. Existing information systems along with data-mining tools can provide better knowledge on the distribution of patient flows. We used one year data of the French Diagnosis Related Groups (DRGs) based system to perform our analysis. Formal Concept Analysis has been used to build Iceberg Lattices of cancer patient flows in the French region of Lorraine. This unsupervised conceptual clustering method allowed us to describe patients flows with an easily understandable visual representation. PMID- 17108602 TI - Non Aristotelian categories in medicine. AB - This paper discusses the representation of medical categories that can not be defined in Aristotelian sense. Two kinds of these categories are mentioned: the prototype and the family resemblance categories. Such categories obviously do exist in medical domain. Search on the Net was performed for free text definition for some commonly used medical categories, like 'autism', 'Burkitt lymphoma' and 'disease'. Most of the found often contradicting definitions do not follow the Aristotelian rules of definition. Many definitions describe statistical properties of the category that are often useless in individual cases. A simple way is suggested that makes possible to represent such categories in biomedical ontologies and treat them separate from better formed categories. This makes possible to revise these categories at any later stage of ontology development. PMID- 17108603 TI - Referent tracking: the problem of negative findings. AB - The paradigm of referent tracking is based on a realist presupposition which rejects so-called negative entities (congenital absent nipple, and the like) as spurious. How, then, can a referent tracking-based Electronic Health Record deal with what are standardly called 'negative findings'? To answer this question we carried out an analysis of some 748 sentences drawn from patient charts and containing some form of negation. Our analysis shows that to deal with these sentences we need to introduce a new ontological relationship between a particular and a universal, which holds when no instance of the universal has a specific qualified ontological relation with the particular. This relation is found to be able to accommodate nearly all occurrences of negative findings in the examined sample, in ways which involve no reference to negative entities. PMID- 17108604 TI - Enriching medical terminologies: an approach based on aligned corpora. AB - Medical terminologies such as those in the UMLS are never exhaustive and there is a constant need to enrich them, especially in terms of multilinguality. We present a methodology to acquire new French translations of English medical terms based on word alignment in a parallel corpus - i.e. pairing of corresponding words. We automatically collected a 27.7-million-word parallel, English-French corpus. Based on a first 1.3-million-word extract of this corpus, we detected 10,171 candidate French translations of English medical terms from MeSH and SNOMED, among which 3,807 are new translations of English MeSH terms. PMID- 17108605 TI - Restructuring the foundational model of anatomy. AB - The authors present a method to convert the FMA to a description logic-based representation in OWL. The concepts denoting anatomical structures are aligned to the DOLCE formal top-level ontology, and converted to a compact core ontology in the spirit of GALEN. The paper presents the identified problems in the FMA and the main aspects of the re-modelling. PMID- 17108606 TI - Four ontological models for radiological diagnostics. AB - In this paper we isolate four diagnostic models in radiology and define a set of diagnostic relations corresponding to each clinical situation. To achieve this, we describe a set of general formal ontological notions, as well as the ontological model of the imaging domain we employed in our analysis. On the basis of our results, we conclude that these diagnostic models and the relations contained therein could be applied to diagnostic situations outside of radiology as well. PMID- 17108607 TI - The derives_from relation in biomedical ontologies. AB - This paper is part of the ongoing efforts within the framework of the Open Biomedical Ontologies (OBO) ontology library to optimize the treatment of relations in biomedical informatics. When ovum and sperm fuse to form a zygote, then the latter is derived from the former. This derives_from relation is of huge importance not only in embryology but also in every other science dealing with ontogenetic processes. This study examines critically the treatment of derives_from in the OBO Relation Ontology in light of its conformity to the underlying processes of development and transformation in biological and medical reality. PMID- 17108608 TI - Tools for Czech biomedical ontologies creation. AB - The paper describes procedures and a tool we have developed to simplify and speed up creating of Czech biomedical ontologies. Our method is based on searching for concepts in a corpus of medical texts and binding those concepts to an established international ontology. The new ontology will have two major advantages: it will be compatible with the international ontology and it will possibly cover all concepts used in the Czech healthcare. The tool supports an author of ontology by mechanizing some routine tasks that occurs in the process of an ontology creation. It tries to learn how to identify concepts in texts and how to bind them to the ontology. The tool then displays the suggestions to a user, who can correct them and add some new ones. Based on this feedback the tool adjusts rules for concept finding and binding. To accomplish such behaviour we have employed some natural language processing methods and information extraction tools. PMID- 17108609 TI - Aligning biomedical ontologies using lexical methods and the UMLS: the case of disease ontologies. AB - The process of aligning ontologies comprises two major steps: i) mapping concepts and ii) characterizing the relations between the concepts. In this paper, we present an alignment method based on a hybrid approach that reuses the UMLS knowledge base and aims at identifying patterns to characterize the relations. The proposed method consist in four steps: 1) exact matching, 2) searching for terms from one ontology that are included in terms from the other ontology, 3) identifying direct relations through the UMLS and 4) extracting syntactico semantic patterns to infer novel alignments. This method has been applied to aligning the Human Disease ontology and the Mouse Pathology ontology resulting in 48 exact matches and 3,697 pairs of concepts for which one term is included in a term from the other ontology. 1,270 alignments are present in the UMLS. Among these, 903 are characterized by a semantic attribute. Based on these alignments, a study of the syntactic patterns has been done. Not surprisingly, the distribution of the different syntactic patterns is not sufficient to discriminate the different types of relationships found in the UMLS alignments. We have used the semantic categorization of the concepts provided by the UMLS to extract syntactico-semantic patterns. 87 novel alignments based on 6 syntactico semantic patterns associated with isa and has associated morphology have been inferred. PMID- 17108610 TI - Towards automated classification of intensive care nursing narratives. AB - Nursing narratives are an important part of patient documentation, but the possibilities to utilize them in the direct care process are limited due to the lack of proper tools. One solution to facilitate the utilization of narrative data could be to classify them according to their content. In this paper, we addressed two issues related to designing an automated classifier: domain experts' agreement on the content of the classes into which the data are to be classified, and the ability of the machine-learning algorithm to perform the classification on an acceptable level. The data we used were a set of Finnish intensive care nursing narratives. By using Cohen's kappa, we assessed the agreement of three nurses on the content of the classes Breathing, Blood Circulation and Pain, and by using the area under ROC curve (AUC), we measured the ability of the Least Squares Support Vector Machine (LS-SVM) algorithm to learn the classification patterns of the nurses. On average, the values of kappa were around 0.8. The agreement was highest in the class Blood Circulation, and lowest in the class Breathing. The LS-SVM algorithm was able to learn the classification patterns of the three nurses on an acceptable level; the values of AUC were generally around 0.85. Our results indicate that one way to develop electronic patient records could be tools that handle the free text in nursing documentation. PMID- 17108611 TI - Comparison of ICHI and CCAM basic coding system. AB - While diagnoses are coded by ICD across the world, there is no universally accepted coding system for procedures. In many countries there exists not even a local classification for medical procedures. As a possible solution the International Classification of Health Interventions (ICHI) has been proposed as a common denominator for an international procedure classification. We alternatively postulate a multiaxial framework for procedure classification following the French Classification Commune des Actes Medicaux (CCAM) for the generation of a procedure shortlist. We compared ICHI and CCAM Basic Coding System focusing on the appropriateness of both systems for supporting the comparability of procedure data. Considering the ongoing standardization of health terminologies and classifications, we strongly recommend to improve the ICHI structure, capitalizing on the benefits of the CCAM architecture. PMID- 17108612 TI - ClaML: a standard for the electronic publication of classification coding schemes. AB - This paper proposes a number of revisions to CEN/TS 14463 (ClaML), which is a pre standard mark-up language for the electronic publication of classification coding schemes. A CEN Taskforce in close collaboration with the WHO network carefully analysed 70 classifications from the healthcare domain. All were transformed in ClaML using a dedicated classification management tool. The proposal removes all formatting elements and adds a number of layout structuring elements. Several elements have been replaced by attributes to enforce internal consistency. A modest number of extensions are proposed to help users and authors in maintenance and version control. A pilot implementation has shown that ICD10 as one of the most complex traditional classifications can be adequately represented to produce quality printed output. PMID- 17108613 TI - The International Classification of Primary Care (ICPC-2): an essential tool in the EPR of the GP. AB - The International Classification of Primary Care (ICPC) has become a standard all over the world. It became a standard tool to classify the important elements in the Electronic Patient Record (EPR) of the GP: reasons for encounter (RFE) reflecting the patient's view, process of care (decision, action, intervention or plans) reflecting the care process, and the assessment (diagnosis or health issue) reflecting the doctor's view. ICPC-2 is fully compatible with structuring data in the episode of care model and it's reflecting the essential elements of each patient/provider encounter. To implement ICPC-2 in the EPR a Thesaurus has been developed in Belgium with double encoded clinical labels. The implementation is now mandatory for labeled EPR systems in Belgium. The use of ICPC 2 ay improve the accessibility and use of on-line Expert systems and Guidelines. PMID- 17108614 TI - Using SNOMED CT codes for coding information in electronic health records for stroke patients. AB - For a project on development of an Electronic Health Record (EHR) for stroke patients, medical information was organised in care information models (templates). All (medical) concepts in these templates need a unique code to make electronic information exchange between different EHR systems possible. When no unique code could be found in an existing coding system, a code was made up. In the study presented in this article we describe our search for unique codes in SNOMED CT to replace the self made codes. This to enhance interoperability by using standardized codes. We wanted to know for how many of the (self made) codes we could find a SNOMED CT code. Next to that we were interested in a possible difference between templates with individual concepts and concepts being part of (scientific) scales. Results of this study were that we could find a SNOMED CT code for 58% of the concepts. When we look at the concepts with a self made code, 54.9% of these codes could be replaced with a SNOMED CT code. A difference could be detected between templates with individual concepts and templates that represent a scientific scale or measurement instrument. For 68% of the individual concepts a SNOMED CT could be found. However, for the scientific scales only 26% of the concepts could get a SNOMED CT code. Although the percentage of SNOMED CT codes found is lower than expected, we still think SNOMED CT could be a useful coding system for the concepts necessary for the continuity of care for stroke patients, and the inclusion in Electronic Health Records. Partly this is due to the fact that SNOMED CT has the option to request unique codes for new concepts, and is currently working on scale representation. PMID- 17108615 TI - SNOMED CT in multidisciplinary clinical practice--evaluation of usefulness for classification and coding of care-planning procedures. AB - Studies of the usefulness of SNOMED CT in everyday clinical practice have received little attention. The purpose of this study was to evaluate the usefulness of SNOMED CT in a multidisciplinary clinical setting. Another aim was to examine how well the terminology of care-planning procedures in a local development project could be mapped to SNOMED CT, and to explore the terminological surroundings in terms of superordinate concepts, subordinate concepts and coordinate concepts. A term list of 66 non-analysed types of procedures, considered necessary in the documentation of care planning, had been compiled in a local development project. The Clue-browser, version 5.5, was used to browse for matching concepts in SNOMED CT, and the degree of correspondence was assessed for each item. Of the 66 procedures, 72% corresponded completely and 22% corresponded partly to concepts in SNOMED CT. No match was found for 6% of the procedures. More subordinate concepts) were lacking in 30% of the concepts. The study showed that SNOMED CT could be mapped fairly well to care-planning procedures in a multidisciplinary setting such as advanced home health care. The terminological surroundings were extensive and stable for superordinate concepts (stem concepts), but unstable regarding subordinate concepts. SNOMED CT and the Clue-browser were feasible for choosing terminology with machine-readable codes for a specific area where there is no terminology in Swedish. PMID- 17108616 TI - Mapping of the WHO-ART terminology on Snomed CT to improve grouping of related adverse drug reactions. AB - The WHO-ART and MedDRA terminologies used for coding adverse drug reactions (ADR) do not provide formal definitions of terms. In order to improve groupings, we propose to map ADR terms to equivalent Snomed CT concepts through UMLS Metathesaurus. We performed such mappings on WHO-ART terms and can automatically classify them using a description logic definition expressing their synonymies. Our gold standard was a set of 13 MedDRA special search categories restricted to ADR terms available in WHO-ART. The overlapping of the groupings within the new structure of WHO-ART on the manually built MedDRA search categories showed a 71% success rate. We plan to improve our method in order to retrieve associative relations between WHO-ART terms. PMID- 17108617 TI - Knowledge acquisition for computation of semantic distance between WHO-ART terms. AB - Computation of semantic distance between adverse drug reactions terms may be an efficient way to group related medical conditions in pharmacovigilance case reports. Previous experience with ICD-10 on a semantic distance tool highlighted a bottleneck related to manual description of formal definitions in large terminologies. We propose a method based on acquisition of formal definitions by knowledge extraction from UMLS and morphosemantic analysis. These formal definitions are expressed with SNOMED International terms. We provide formal definitions for 758 WHO-ART terms: 321 terms defined from UMLS, 320 terms defined using morphosemantic analysis and 117 terms defined after expert evaluation. Computation of semantic distance (e.g. k-nearest neighbours) was implemented in J2EE terminology services. Similar WHO-ART terms defined by automated knowledge acquisition and ICD terms defined manually show similar behaviour in the semantic distance tool. Our knowledge acquisition method can help us to generate new formal definitions of medical terms for our semantic distance terminology services. PMID- 17108618 TI - Construction of a semi-automated ICD-10 coding help system to optimize medical and economic coding. AB - INTRODUCTION: In order to measure the medical activity in hospitals, physicians are required to code manually information concerning a patient's stay using ICD 10. This requires trained staff and a lot of time. We propose to help speed up and facilitate the tedious task of coding patient information. METHODS: we show two methods. First, we propose an automated ICD-10-based coding help system using an automated MeSH-based indexing system and a mapping between MeSH and ICD-10 extracted from the UMLS metathesaurus. Secondly, we propose the use of drug prescriptions to complete the previous coding with the use of a mapping between a given prescription drug and the relevant ICD-10 codes (in compliance with the drug approval). RESULTS: the results of a preliminary experiment indicate that the precision of the indexing system is 40% and the recall is 30% when we compare to an economic rules-based coding and to a descriptive coding. DISCUSSION: moreover, we show that the use of prescription coding is relevant as the recall reaches 68% when the Vidal tool is used. CONCLUSION: Then, it is very interesting to complete the coding obtained automatically by the indexing/mapping system by the coding obtained from the prescriptions. PMID- 17108619 TI - Interactive visualization and navigation of complex terminology systems, exemplified by SNOMED CT. AB - Free-text queries are natural entries into the exploration of complex terminology systems. The way search results are presented has impact on the user's ability to grasp the overall structure of the system. Complex hierarchies like the one used in SNOMED CT, where nodes have multiple parents (IS-A) and several other relationship types, makes visualization challenging. This paper presents a prototype, TermViz, applying well known methods like "focus+context" and self organizing layouts from the fields of Information Visualization and Graph Drawing to terminologies like SNOMED CT and ICD-10. The user can simultaneously focus on several nodes in the terminologies and then use interactive animated graph navigation and semantic zooming to further explore the terminology systems without loosing context. The prototype, based on Open Source Java components, demonstrates how a number of Information Visualisation methods can aid the exploration of medical terminologies with millions of elements and can serve as a base for further development. PMID- 17108620 TI - Cross-lingual alignment of biomedical acronyms and their expansions. AB - We propose a method that aligns biomedical acronyms and their definitions across different languages. The approach is based upon a freely available tool for the extraction of abbreviations together with their expansions, and the subsequent normalization of language-specific variants, synonyms, and translations of the extracted acronym definitions. In this step, acronym expansions are mapped onto a language-independent concept-layer on which intra- as well as interlingual comparisons are drawn. PMID- 17108621 TI - An Ontology driven collaborative development for biomedical terminologies: from the French CCAM to the Australian ICHI coding system. AB - The CCAM French coding system of clinical procedures was developed between 1994 and 2004 using, in parallel, a traditional domain expert's consensus method on one hand, and advanced methodologies of ontology driven semantic representation and multilingual generation on the other hand. These advanced methodologies were applied under the framework of an European Union collaborative research project named GALEN and produced a new generation of biomedical terminology. Following the interest in several countries and in WHO, the GALEN network has tested the application of the ontology driven tools to the existing reduced Australian ICHI coding system for interventions presently under investigation by WHO to check its ability and appropriateness to become the reference international coding system for procedures. The initial results are presented and discussed in terms of feasibility and quality assurance for sharing and maintaining consistent medical knowledge and allowing diversity in linguistic expressiveness of end users. PMID- 17108622 TI - The redesign of the Medical Informatics Master of Science course at the University of Amsterdam. AB - OBJECTIVES: To describe our new two years Master of Science (MSc) program starting in September 2006 at the University of Amsterdam- Academic Medical Center, The Netherlands. METHODS: We elaborate shortly on the mission, organizational structure and new contents of this new MSc course in medical informatics. RESULTS: Through the years, our medical informatics university program underwent some major revisions of which the transition from a four years course into a three years BSc program and a two years MSc program has been the most fundamental. The new MSc program is aimed at (international) baccalaureates in medical informatics, computer science, medicine, health sciences, and biology. Besides, health care professionals or professionals with a background in computer science may enter the program. The program length is two years, comprising four study semesters of 30 European Credits each (EC, 1 EC corresponding to 27 hours study load), equalizing 120 EC in total of which 48 EC are reserved for the master's thesis. CONCLUSIONS: With the new set up of the MSc program, that will be offered in English, we hope to both accommodate the learning needs of our own baccalaureates and to attract international baccalaureates and other professionals to this course. Our ultimate aim is to bring forth medical informatics specialists who are well equipped to make significant contributions to the field. PMID- 17108623 TI - Medical education and role of medical informatics. AB - Bosnia and Herzegovina (BiH), as developing country in transition, has to shift from traditional ways of learning to the transformation of the university education in accordance with Bologna process and educational standards in European Union. In the light of these changes authors conducted research at bio medical faculties in Sarajevo in order to address issues of the education of future physicians and especially role of medical informatics in their under and post graduate studies and continuous medical education. As per given results in this study, current quality of medical education at biomedical faculties, University of Sarajevo, is unsatisfactory due to several reasons and some among others are those traditionally seen as "computer literacy". Problems are determined and recommendations are given for decision makers to support transformation of BiH medical educational system to have physicians, dentists, pharmacists and nurses who possess the knowledge, skills and attitudes required to be competent in medical informatics if they wish to incorporate into their practices systematic approaches for promoting and maintaining the health of defined populations. PMID- 17108624 TI - Developing an interactive approach in teaching medical informatics. AB - A new masters program in medical informatics is proposed for development at the University of Medicine and Pharmacy in Timisoara. Given the rapidly changing technology itself and its deployment in biomedical science, the master's program curriculum has to be multidisciplinary, comprehensive and coherent in conveying the concepts, as well as the interdisciplinary character, of medical informatics (MI). We describe the rationale and methods for a pilot study to develop a new, interactive approach in teaching MI. The study is being conducted within the existing MI course offered for the medical students in order to evaluate its impact on instruction and determine if a larger scale design is feasible. Two teaching teams of four instructors have been assigned to one of two tracks in our pilot study: traditional instruction or interactive instruction. After one term we have gained important information about how the structural and instructional aspects of the pilot design may influence confidence and attitudes. PMID- 17108625 TI - The INFOBIOMED Network of Excellence: facilitating training and mobility in biomedical informatics in Europe. AB - In this paper we present the work carried out within INFOBIOMED, the European Network of Excellence (NoE) in Biomedical Informatics, to facilitate training and mobility in the area. An analysis of past experiences in both Medical Informatics and Bioinformatics has led to various actions. In this scenario, we have elaborated three actions: (1) a survey of the training and mobility situation, needs and actions to be taken, (2) a Biomedical Informatics course database (ICD), and (3) a Mobility Brokerage Service (MBS). We describe the use of Web Services to build the MBS, designed to facilitate the exchange of professionals within the consortium, belonging to ten European countries. The goal of INFOBIOMED is to expand these initiatives to other NoEs and institutions within the European Union. PMID- 17108626 TI - Multimedia and physiology: a new way to ensure the quality of medical education and medical knowledge. AB - BACKGROUND: since the eighties and the existence of virtual campuses, the value of computers in distance education has been acknowledged. The development of information and communication technologies is driving at discriminating distance education and on-line education. PURPOSE: the aim of the "Campus Numerique de Physiologie" is not to reproduce an on-line copy of classical textbooks but to put at students' and physicians' disposal the huge possibilities of multimedia resources for an active and easier understanding of complex physiopathological phenomena. METHODS: the on-line course materials were created using both original IBC-made and registered trade-mark software tools. Multiscale modelling and corresponding knowledge bases were implemented by mathematicians, biologists and software engineers from Rennes. The website, which is accessible through a server of the French Virtual Medical University, was developed in the language HTML/PHP connected to a MySQL database. RESULTS: the content managing system is consistent with classical home page facilities and multicriteria browser. Interactive resources are freely available for the site's users. Two- and three-dimensional simulations born out of mathematical qualitative and quantitative models at the molecular, cellular or organic level keep students active with regards to fundamental mechanisms by interactively manipulating the simulation environment. CONCLUSION: authors comment the already available course materials which should stimulate the creation of new documents following a validation by a qualified commission of the "Societe de Physiologie". Providing evaluation tests, teachers anticipate that the increasing content of this virtual campus will allow users to gain a complete understanding and an integrative view of many physiopathological mechanisms. PMID- 17108627 TI - ENN-ICS--implementation and evaluation of a multilingual learning management system for sleep medicine in Europe. AB - A new web based network aims at the improvement of health care in Europe by integrating advanced e-learning and e-publishing technologies for the training of medical doctors, nurses, and students. The field of application is sleep physiology and sleep medicine. Based on a multilingual, multimedia communication system, ENN-ICS Centre offers direct access to medical information for users, i.e healthcare professionals and citizens, in Europe and worldwide. The use of XML supports the development of media independent contents for multiple target groups. Editorial and distributive processes are supported by customized central editorial, content management and learning management systems (CMS, LMS). ENN-ICS e-health services are evaluated by selected user groups in North, Middle and Southern Europe using reliable and scientifically accepted validation instruments. The compliance with essential quality requirements and criteria is tested and verified by using online questionnaires based on the DISCERN questionnaire for evaluating patient information, the HON principles for health related websites and the GMDS catalogue of quality criteria for electronic publications in medicine. The system architecture and its exemplary applications can be used as a model for future e-health services dealing with neurological and other medical topics. PMID- 17108628 TI - What makes an "informed patient"? The impact of contextualization on the search for health information on the Internet. AB - The Internet is nowadays a powerful medium and can help patients to become better informed citizens. Increasingly, citizens are searching for health information on the Internet. The Internet-based resource often acts as a virtual healthcare professional. The effectiveness of the communication between the patient and the virtual healthcare professional depends partly upon the presence of contextual information. The issue here is to what extent the contextualization of information is needed for effective information seeking and for the person's understanding of the retrieved/received information. The impact of contextualization on information search also closely relates to the person's cognitive resources. Using a theoretical communication framework (Te'eni 2001), we explore contextualization in a health website, and discuss the above issues and the possible relevance of contextualized information on the informed patient. PMID- 17108629 TI - The effect of a multimedia health educational program on the postoperative recovery of patients undergoing laparoscopic cholecystectomy. AB - Aim of this study is the evaluation of the impact of preoperative informative session using a Multimedia Health Educational Program (MHEP) on patients undergoing elective Laparoscopic Cholecystectomy (LC) for cholelithiasis, preoperative anxiety and postoperative pain and nausea. Sixty consecutive patients scheduled for elective LC were considered for enrollment in the trial. Patients were assigned randomly to four groups: Group A included 15 patients, preoperatively informed regarding LC through the MHEP presented by a Registered Nurse (RN). Group B included 15 patients preoperatively informed through a leaflet (designed and developed using the exact contents of the MHEP). In Group C, there were 15 patients who were being informed verbally from the RN. Finally, the control Group D included 15 patients, who had the conventional preoperative information about the operation and postoperative course by the attending surgeon and anesthesiologist, as every other patient included in groups A, B, C. Preoperative assessment of patient's knowledge about cholelithiasis and LC was performed after informative session, and was based on a specifically developed "closed, true-false" questionnaire. Preliminary results suggest that conventional information provided by the attending surgeon (Group D) is inadequate. Specifically developed informative sessions with the contribution of MHEP seems to be effective on reducing preoperative anxiety and postoperative pain, in patients undergoing elective LC. PMID- 17108630 TI - A new health strategy to prevent pressure ulcer formation in paraplegics using computer and sensory substitution via the tongue. AB - Pressure ulcers are recognized as a major health issue in individuals with spinal cord injuries and new approaches to prevent this pathology are necessary. An innovative health strategy is being developed through the use of computer and sensory substitution via the tongue in order to compensate for the sensory loss in the buttock area for individuals with paraplegia. This sensory compensation will enable individuals with spinal cord injuries to be aware of a localized excess of pressure at the skin/seat interface and, consequently, will enable them to prevent the formation of pressure ulcers by relieving the cutaneous area of suffering. This work reports an initial evaluation of this approach and the feasibility of creating an adapted behavior, with a change in pressure as a response to electro-stimulated information on the tongue. Obtained during a clinical study in 10 healthy seated subjects, the first results are encouraging, with 92% success in 100 performed tests. These results, which have to be completed and validated in the paraplegic population, may lead to a new approach to education in health to prevent the formation of pressure ulcers within this population. PMID- 17108631 TI - Reducing dropouts in outpatient care through an SMS-based system. AB - The objective of this work is to reduce the number of outpatients dropouts, i.e. appointments that, unexpectedly, are not attended by the scheduled patients, thus causing disturbances to the regular ambulatory workflow, waste of resources, and, eventually, longer waiting lists for ambulatory care. A collateral, but not less important result is the improvement of the relationship between citizens and the public Hospital Company. The method proposed consists in sending short instant messages to remind the appointment. After the promising estimates obtained by a preliminary cost-benefit analysis, a commercial product was purchased for the communication infrastructure, while a software module has been developed for interacting with the legacy system, in order to retrieve the information necessary to compose the text of every sms and send it to the citizen. The system has been implemented since seven months, and its benefits have been measured. Dropouts trend, stable at 7-8% during last years, is clearly decreasing, while the number of citizens providing their mobile number to the healthcare booking facilities is constantly increasing. Our conclusion is that the cost of the system will be amortised in a very short time, leading to a significant cost saving and, hopefully, to shorter waiting lists. PMID- 17108632 TI - Evaluation of a discussion forum for knowledge sharing among emergency practitioners: a social network approach. AB - Peer to peer knowledge sharing is recognized as a key contributor to the development of expert practice for health care professionals. Emergency departments with access to extensive expertise, such as in urban hospital settings, present greater potential for rich collaborative learning opportunities as compared with rural settings where expertise is at times scarce. Collaborative technologies such as electronic discussion boards may assist in leveling the "knowledge" playing field and increase opportunities for the growth of a strong social network for emergency clinicians. A social network perspective is used to explore the effectiveness of a discussion forum to support knowledge sharing among emergency practitioners in rural and urban emergency departments in Nova Scotia. PMID- 17108633 TI - On neural network classification of otoneurological cases on the basis of recognition results of vestibulo-ocular reflex eye movements signal. AB - We constructed a signal analysis and recognition system to compute revealing results from vestibulo-ocular reflex eye movements and their stimulation head movements of impulsive type that were generated as passive head movements with a special device designed by us for this purpose. Further, we implemented perceptron neural networks to separate healthy subjects from patients suffering from dysfunction of vestibulo-ocular reflex in either ear. We gained high classification accuracies with this method ready for routine use. PMID- 17108634 TI - Publication bias in medical informatics evaluation research: is it an issue or not? AB - The phenomenon of publication bias has probably existed since results of scientific research are being published. Positive and/or statistically significant results seem more likely to be published than negative and/or insignificant results. However, it is unclear if there is a remarkable impact of publication bias in medical informatics evaluation literature and how aware researchers are of its effect. We conducted a small-scale study in order to find out what the ratio of papers describing positive results vs. negative results is, tried to find enough studies to a certain subject to carry out a meta-analysis and assess publication bias by statistical methods, and finally examined reviews and meta-analyses for their results and their quality. A random sample of 86 studies showed a remarkably high percentage of descriptions of positive results (69.8%). 19 (36.6%) of the analyzed 54 reviews and meta-analyses came to a positive conclusion with regard to the overall effect of the analyzed system, 32 (62.5%) were inconclusive, and only one review came to a negative conclusion. Quantitative assessment of publication bias for health informatics studies was found difficult due to the low number of comparable studies. Although there is no clear evidence for a great impact of publication bias in medical informatics evaluation literature, further research should carried out. PMID- 17108635 TI - The multiple faces of the e-patient, if not disabled. AB - The term e-health entered common speech a few years ago , while the term "e patient" has recently appeared on the healthcare scenario. The aim of the present paper is to describe the nature of the so called "e-patient" from different points of view, through a review, not systematic, of the literature. A profile, though not totally exhaustive, of the current e-patient has been drawn, in an attempt to report uncertainties and worries that should not be underestimated. Comments are provided on the asymmetry between the evaluation effort around the internet world and corresponding face-to-face world. Disabled patients are almost excluded from the e-patient family due to the inaccessibility of most of the health related web sites. PMID- 17108636 TI - Computing latent taxonomies from patients' spontaneous self-disclosure to form compatible support groups. AB - A growing number of Internet sites provide patients with an opportunity to share experiences about their illness. Unfortunately, it depends on the luck of the draw whether patients find compatible others for mutual support. To improve this situation, this paper demonstrates how tools from Information Retrieval can be deployed to discover patients with compatible stories more systematically. We show how finding compatible support is like finding relevant documents on the Internet, and also in what way it is different. We designed an algorithm to produce a 'latent' taxonomy over patients' self-disclosures to aid them in forming support groups. At the end we briefly consider the result of this taxonomy based on only the patients' stories, with one based on explicit questionnaires. PMID- 17108637 TI - Generation of 4D CT image data and analysis of lung tumour mobility during the breathing cycle. AB - The mobility of lung tumours during the breathing cycle is a source of error in radiotherapy treatment planning. Spatio-temporal CT data sets can be used to measure the movement of lung tumours caused by breathing. Because modern CT scanners can only scan a limited region of the body simultaneously at different times, patients have to be scanned in segments consisting of multiple slices. For studying free breathing motion multislice CT scans can be collected simultaneously with digital spirometry over several breathing cycles. The image data set is assembled by sorting the free breathing multislice CT scans according to the couch position and the tidal volume. But artefacts can occur because there are no data segments for exactly the same tidal volume and all couch positions. In this paper, a non-linear registration method is used to interpolate and reconstruct 4D CT data sets from multislice CT scans in high quality. The non linear registration estimates a velocity field between successive scans, which is used to reconstruct a 4D CT data set by interpolating data at user-defined tidal volumes. By this technique, artefacts can be reduced significantly. Furthermore, the reconstructed 4D CT data sets are used for studying the motion of lung tumours during the respiratory cycle. The reconstructed 4D data sets of 4 patients were used to quantify the individual lung tumour motion as well as to estimate the tumour's appearance probability during a breathing cycle. PMID- 17108638 TI - Kernel methods for melanoma recognition. AB - Skin cancer is a spreading disease in the western world. Early detection and treatment are crucial for improving the patient survival rate. In this paper we present two algorithms for computer assisted diagnosis of melanomas. The first is the support vector machines algorithm, a state-of-the-art large margin classifier, which has shown remarkable performances on object recognition and categorization problems. The second method, spin glass-Markov random fields, combines results of statistical physics of spin glasses with Markov random fields. We compared the two approaches using color histograms as features. We benchmarked our methods with another algorithm presented in the literature, which uses a sophisticated segmentation technique and a set of features especially designed for melanoma recognition. To our knowledge, this algorithm represents the state of the art on skin lesions classification. We show with extensive experiments that the support vector machines approach outperforms the existing method and, on two classes out of three, it achieves performances comparable to those obtained by expert clinicians. PMID- 17108639 TI - Pre- and intraoperative processing and integration of various anatomical and functional data in neurosurgery. AB - A software system is presented, capable of integrating various information sources for neurosurgical procedures. These include anatomical data such as a standard 3D DICOM image stacks, atlas data, as well as functional information (e.g. fmri, MEG, EEG). The system is programmed in C++ using Open GL for visualisation, and was developed in a close cooperation with a neurosurgical department to match existing needs. Preoperatively the data may be combined, registered by a rigid or elastic matching process, enriched by user specified planning information such as annotations or trajectories, and visualised in a standard fashion using different segmentation schemes, interactive rotation, zooming etc. Selected portions of the gathered and generated information may then be exported for neuronavigation input in DICOM or a vendor specific format. Intraoperatively, this information may, on the one hand, be simply used as an integrational part of the routinely used navigational data. On the other hand, the system is also capable of interacting with the navigational system to integrate the actual spatial information of the ongoing procedure into the preoperative data, thus allowing further planning and visualisation beyond the scope of the navigational unit. Furthermore, intraoperatively updated information such as intraoperative MR images or electrophysiological data may be integrated and correlated to the existing information. Ongoing developments comprise redistribution of the relevant data for injection onto the screen of the navigational system or into the optical pathway of the (3D capable) microscope display/ocular. This also includes information about the actual automatically optimized accuracy of the navigation process in relation to the head markers in use. PMID- 17108640 TI - Progressing professional maturity in health informatics. AB - This paper looks at ways in which the profession of health informatics is gaining definition, through the actions of both the BCS Health Informatics Forum and the UK Council for Health Informatics Professions. It explores the some of the ways the current situation was achieved, how the lessons can be applied internationally and the challenges yet to be met. Opportunities for further progress within the existing landscape of technology and health care delivery and management are outlined. The paper documents actions taken and planned and the rationale of why, with supporting evidence of similar contributory work to date. It concludes by stating that the profession must remain flexible and dynamic, and nurture its professional members if it is to continue to develop within an ever changing scenario. PMID- 17108641 TI - Bridging information gaps between primary and secondary healthcare. AB - Medication errors are harmful and costly for healthcare systems. Recent studies show that a major part of these errors are due to the problems in transferring the patient current medication-data between primary and secondary healthcare. Recent ICT development promises to improve the communication between primary and secondary healthcare. In order to find out the constraints that may hamper a communication project's productivity, an IT configuration for building a medication-data communication network between primary and secondary healthcare in the Netherlands was followed applying qualitative methods. We analysed some important problems that project faced and conclude that problems with the data integration and saving the data integrity are important challenges for the project to maintain its objectives. PMID- 17108642 TI - The demographic bias of email as a survey method in a pediatric emergency population. AB - Email has been considered as a communication medium between patients and clinicians in pediatric emergency departments, but the demographic bias involved in using email has not been fully explored. We developed a paper based survey to explore access, willingness to participate and the demographic bias of email within our parent population. METHODS: To 1733 possible subjects, 1200 surveys were distributed with a return of 1018, a survey response rate of 85%, and a population response rate of 59%. RESULTS: Subjects from families with incomes less than $60,000 per year had lower access rates (OR = 0.40, 95 (OR = 0.37, [0.17, 0.81]). Employment outside of the home was associated with increased email access rates (OR = 1.79, 95% CI [1.19, 2.70]). Visible minority status was associated with an increased willingness to participate (OR = 1.84, 95 as was low education (OR = 2.12, 95% CI [1.04, 4.32]). The population of theoretical responders to an email based quality assurance process would have been significantly different from the base population of adults accompanying children to our emergency department as a result of these biases. CONCLUSIONS: We have demonstrated a degree of demographic bias in email access rates, negatively affecting those individuals with lower income, less employment, and lower education. Email based surveys directed at parents in pediatric emergency departments should include questions on income, employment and education in order to permit those who analyze the data to correct for these variables. More research is needed to confirm these findings. PMID- 17108643 TI - Artificial neural network versus subjective scoring in predicting mortality in trauma patients. AB - OBJECTIVE: Current methods of trauma outcome prediction rely on clinical knowledge and experience. This makes the system a subjective score, because of intra-rater variability. This project aims to develop a neural network for predicting survival of trauma patients using standard, measured, physiological variables, and compare its predictive power with that obtained from current trauma scores. METHODS: The project uses 7688 patients admitted to the Swedish Medical Center, Colorado, U.S.A. between the years 2000-2003 inclusive. Neural Network software was used for data analysis to determine the best network design on which to base the model to be tested. The model is created using a minimum number of variables to produce an effective outcome predicting score. Initial variables were based on the current variables used in calculating the Revised Trauma Score, replacing the Glasgow Coma Scale (GCS) with a modified motor component of the GCS. Additional variables are added to the model until a suitable model is achieved. RESULTS: The best model used Multi-Layer Perceptrons, with 8 input variables, 5 hidden neurons and 1 output. It was trained on 5881 cases and tested independently on 1807 cases. The model was able to accurately predict 91% patient mortality. CONCLUSIONS: An ANN developed using pre-hospital physiological variables without using subjective scores resulted in good mortality prediction when applied to a test set. Its performance was too sensitive and requires refinement. PMID- 17108644 TI - Clinical pathways development and computer support in the EPR: lessons learned. AB - This paper refers to a project for development and optimization of a clinical pathway for the Lumbar Nerve Root Compression Syndrome. A special focus is taken on computer support for pathway development and implementation. An innovative combination of "rapid prototyping" of a workflow model and a 2 level approach using round robin methods in a large group and individual semi-structured interviews is presented. The method focuses on process optimization instead of process modeling and concentrates on areas of the workflow which may be optimized. Critical parts of the optimized clinical workflow have been implemented inside a commercial electronic patient record system. PMID- 17108645 TI - Removal of paper-based health records from Norwegian hospitals: effects on clinical workflow. AB - Several Norwegian hospitals have, plan, or are in the process of removing the paper-based health record from clinical workflow. To assess the impact on usage and satisfaction of electronic health record (EHR) systems, we conducted a survey among physicians, nurses and medical secretaries at selected departments from six Norwegian hospitals. The main feature of the questionnaire is the description of a set of tasks commonly performed at hospitals, and respondents were asked to rate their usage and change of ease compared to previous routines for each tasks. There were 24 tasks for physicians, 19 for nurses and 23 for medical secretaries. In total, 64 physicians, 128 nurses and 57 medical secretaries responded, corresponding to a response rate of 68%, 58% and 84% respectively. Results showed a large degree of use among medical secretaries, while physicians and nurses displayed a more modest degree of use. Possibly suggesting that the EHR systems among clinicians still is considered more of an administrative system. Among the two latter groups, tasks regarding information retrieval were used more extensively than tasks regarding generating and storing information. Also, we observed large differences between hospitals and higher satisfaction with the part of the system handling regular electronic data than scanned document images. Even though the increase in use among clinicians after removing the paper based record were mainly in tasks where respondents had no choice other than use the electronic health record, the attitude towards EHR-systems were mainly positive. Thus, while removing the paper based record has yet to promote new ways of working, we see it as an important step towards the EHR system of tomorrow. Several Norwegian hospitals have shown that it is possible. PMID- 17108646 TI - Order-disorder transition in monoclinic sulfur: a precise structural study by high-resolution neutron powder diffraction. AB - High-resolution neutron powder diffraction has been used in order to characterize the order-disorder transition in monoclinic cyclo-octasulphur. Rapid data collection and the novel use of geometrically constrained refinements has enabled a direct and precise determination of the order parameter, based on molecular site occupancies, to be made. The transition is critical and continuous; with a transition temperature, Tc=198.4 (3) K, and a critical exponent, beta=0.28 (3), which is indicative of three-dimensional ordering. Difficulties encountered as a consequence of the low thermal conductivity of the sample are discussed. PMID- 17108647 TI - Structure of strontium barium niobate SrxBa1-xNb2O6 (SBN) in the composition range 0.32stishovite transition, GaAsO4 transforms into a dirutile structure at 9 GPa and 1173 K. In 2002, a new GaAsO4 polymorph was found by quenching the compound from 6 GPa and 1273 K to ambient conditions. The powder diagram was indexed on the basis of a hexagonal cell (a=8.2033, c=4.3941 A, V=256.08 A3), but the structure did not correspond to any known structure of other AXO4 compounds. We report here the ab initio crystal structure determination of this hexagonal polymorph from powder data. The new phase is isostructural to beta-MnSb2O6 and it can be described as a lacunary derivative of NiAs with half the octahedral sites being vacant, but it also contains fragments of the rutile-like structure. PMID- 17108656 TI - Theoretical determination of the structures of CaSiO3 perovskites. AB - Density functional theory is used to determine the possible crystal structure of the CaSiO3 perovskites and their evolution under pressure. The ideal cubic perovskite is considered as a starting point for studying several possible lower symmetry distorted structures. The theoretical lattice parameters and the atomic coordinates for all the structures are determined, and the results are discussed with respect to experimental data. PMID- 17108657 TI - Superspace-symmetry determination and multidimensional refinement of the incommensurately modulated structure of natural fresnoite. AB - The structure of natural fresnoite, Ba2TiSi2O8, from the sanbornite deposits of eastern Fresno County, California, has been solved and refined as an incommensurate structure in five-dimensional superspace. The structure is tetragonal, superspace group P4bm(alpha, alpha, 1/2)(-alpha, alpha, 1/2)0gg, cell parameters a=8.5353 (6), c=10.4128 (7) A, modulation vectors q1=0.3020 (3) (a*+b*), q2=0.3020 (3) (-a*+b*). Data collection was performed on an Xcalibur CCD diffractometer at 110 K. The structure was refined from 3452 reflections to final R=0.0123. The model includes modulation of both atomic positions and displacement parameters. As a consequence of the Ba and O positional modulation, eight-, nine- and tenfold Ba coordinations occur throughout the structure. The change of coordination around the Ba atom is clearly represented by the deformation of the pentagonal rings, as seen from a projection along [001]. The deformed pentagonal rings correspond to Ba atoms with eight- and ninefold coordinations and form octagonal clusters closely resembling those observed in the incommensurate structure of melilite-type compounds. PMID- 17108658 TI - A new simplifying approach to molecular geometry description: the vectorial bond valence model. AB - A method to describe, analyze and even predict the coordination geometries of metal complexes is proposed, based on previous well established concepts such as bond valence and valence-shell electron-pair repulsion (VSEPR). The idea behind the method is the generalization of the scalar bond-valence concept into a vector quantity, the bond-valence vector (BVV), with the innovation that the multidentate ligands are represented by their resultant BVVs. Complex n-ligand coordination spheres (frequently indescribable at the atomic level) reduce to much simpler ones when analyzed in BVV space, with the bonus of a better applicability of the VSEPR predictions. The geometrical implications of the BVV description are analyzed for the cases of n=2 and 3 (n=number of ligands), and the validity of its predictions, checked for a large number of metal complexes. PMID- 17108659 TI - Growth-induced incommensurability observed in the organic co-crystal hexamethylenetetramine resorcinol. AB - Co-crystals of hexamethylenetetramine and resorcinol were investigated by X-ray diffraction. The structure was refined in the superspace group Xmcm(0beta0)s0s, X=(1/2 1/2 0 1/2). In the average structure the resorcinol molecules are disordered between two orientations. The main effect of the modulation in the structure is a harmonic modulation of the occupation probabilities of the two orientations of the resorcinol molecule. However, the modulated order is not perfect and the resorcinol molecules remain partially disordered. Below 270 K the crystal undergoes a phase transition to a commensurately modulated structure with modulation vector q=(0 1/2 1/4) and superspace group X2/m(0betagamma)0s. The structure of the low-temperature phase could not be determined owing to the poor quality of the crystals affected by the reconstructive phase transition. PMID- 17108660 TI - Structural characterization of p-benzosemiquinone radical in a solid state: the radical stabilization by a low-barrier hydrogen bond. AB - Semiquinone (p-benzosemiquinone), a transient organic radical, was detected in the solid state by EPR spectroscopy revealing four symmetrically equivalent protons. A variable-temperature X-ray diffraction analysis (293 and 90 K) and EPR data support a dynamical disorder of the proton. A low-barrier O-H...O hydrogen bond stabilizes the radical. The C-O bond length is 1.297 (4) A, corresponding to a bond order of ca 1.5. The geometry of the radical implies an electron delocalization throughout the benzenoid ring. Two polymorphs of semiquinone, monoclinic and triclinic, were observed and their structures determined. Their crystal packings were compared with those of quinhydrone polymorphs. PMID- 17108661 TI - Hydrophobic 'lock and key' recognition of N-4-nitrobenzoylamino acid by strychnine. AB - During racemic resolution of N-4-nitrobenzoyl-DL-amino acids (alanine, serine and aspartic acid) by a fractional crystallization of strychninium salts, crystals of both diastereomeric salts were grown, and the crystal structures of strychninium N-4-nitrobenzoyl-L-alaninate methanol disolvate (1a), strychninium N-4 nitrobenzoyl-D-alaninate dihydrate (1b), strychninium N-4-nitrobenzoyl-D-serinate dihydrate (2a), strychninium N-4-nitrobenzoyl-L-serinate methanol solvate hydrate (2b), strychninium hydrogen N-4-nitrobenzoyl-L-aspartate 3.75 hydrate (3a) and strychninium hydrogen N-4-nitrobenzoyl-D-aspartate 2.25 hydrate (3b) were determined. The strychninium cations form corrugated layers, which are separated by hydrogen-bonded anions and solvent molecules. Common features of the corrugated layers are deep hydrophobic grooves at their surfaces, which are occupied by the 4-nitrobenzoyl groups of suitable anions. The hydrophobic ;lock and key' recognition of 4-nitrobenzoyl groups of amino acid derivatives in deep grooves of the strychnine self-assembly causes the resulting surface to have more hydrophilic properties, which are more appropriate for interactions in the hydrophilic environments from which strychninium salts were crystallized. In the crystal structure of (2a) and (3a), such hydrophobic ;lock and key' recognition is responsible for the lack of N-H+...O- hydrogen bonds that are usually formed between the protonated tertiary amine N atom of the strychninium cation and the deprotonated carboxyl group of the resolved acid. In the crystal structure of (2a) and (3a), the protonated amine N atom is a donor of hydrogen bonds, while the hydroxyl group of the serine derivative and water molecules are their acceptors. In light of the hydrophobic recognition, chiral discrimination depends on the nature of the hydrogen-bond networks, which involve anions, solvent molecules and the protonated amine N atom of strychninium cations. PMID- 17108662 TI - Pressure-frozen 1,2,3-trichloropropane. AB - The structure of 1,2,3-trichloropropane, ClCH2CHClCH2Cl, in-situ crystallized in a diamond-anvil cell, has been determined by single-crystal X-ray diffraction at 0.28 and 0.35 GPa. A melting point at 295 K and 0.22 GPa has been determined. The molecular conformation of aliphatic chain and terminal Cl atoms is approximately C2 symmetric. Of the intermolecular contacts, the tightest are the Cl...Cl and then the Cl...H contacts, whereas the H...H distances are considerably longer than the sum of the van der Waals radii and leave narrow voids in the structure. The elevated pressure reduces the H...H separations but hardly affects the Cl...Cl distances. The crystal growth rates, compression and types of intermolecular interactions in the structures can be correlated with the directions of the Cl...Cl contacts, which can be considered the main motif responsible for the molecular arrangement in the structure. PMID- 17108663 TI - Competing hydrogen-bonding patterns and phase transitions of 1,2-diaminoethane at varied temperature and pressure. AB - 1,2-diaminoethane has been in-situ pressure- and temperature-frozen; apart from two known low-temperature phases, Ialpha and II, three new phases, Ibeta, Igamma and III, have been observed and their structures determined by X-ray diffraction. The measurements at 0.1 MPa were carried out at 274, 243 and 224 K, and 296 K measurements were made at 0.15 GPa (phase Ialpha), at 0.3 and 1.1 GPa (phase Ibeta), at 1.5 GPa (phase Igamma), and at 0.2, 0.3 and 0.5 GPa (phase III). All these phases are monoclinic, space group P2(1)/c, but the unit-cell dimension of phases Ialpha and III are very different at 296 K: aIalpha=5.078 (5), bIalpha=7.204 (8), cIalpha=5.528 (20) A, betaIalpha=115.2 (2) degrees at 0.15 GPa, and aIII=5.10 (3), bIII=5.212 (2), cIII=7.262 (12) A, betaIII=111.6 (4) degrees at 0.2 GPa, respectively; in both phases Z=2. An ambient-pressure low temperature phase II has been observed below 189 K. Discontinuities in the unit cell dimensions and in the N...N distance mark the isostructural transition between phases Ialpha and Ibeta at 0.2 GPa, which can be attributed to a damping process of the NH2 group rotations. In phase Igamma the unit-cell parameter a doubles and Z increases to 4. The molecule has inversion symmetry in all the structures determined. 1,2-Diaminoethane can be considered as a simple structural ice analogue, but with NH...N hydrogen bonds and with the H-atom donors (four in one molecule) in excess over H-atom acceptors (two per molecule). Thus, the transformations of 1,2-diaminoethane phases involving the conformational dynamics affect the hydrogen-bonding geometry and molecular association in the crystal. The 1,2-diaminoethane:1,2-dihydroxyethane mixture has been separated by pressure freezing, and a solid 1,2-diaminoethane crystal in liquid 1,2-dihyroxyethane has been obtained. PMID- 17108664 TI - Molecular interactions in crystalline dibromomethane and diiodomethane, and the stabilities of their high-pressure and low-temperature phases. AB - Dibromomethane, CH2Br2, and diiodomethane, CH2I2, have been in situ pressure crystallized in a diamond-anvil cell and their structures determined by single crystal X-ray diffraction at 0.61 and 0.16 GPa, respectively. The pressure-frozen CH2Br2 crystal is isostructural with its C2/c phase obtained by cooling. CH2I2 is known to form several phases at low temperature, one of which is isostructural with CH2Br2. However, pressure freezing leads to the polar Fmm2 phase. The formation of the polar CH2I2 structure at 0.16 GPa has been rationalized by the electrostatic and anisotropic van der Waals interactions of the I atoms. No ferroelectric behaviour of the Fmm2 polar phase II of CH2I2 has been determined. The diffraction, calorimetric and dielectric constant studies reveal considerable temperature hysteresis of transformations between the CH2I2 phases, as well as metastable regions strongly dependent on the sample shape and history. PMID- 17108665 TI - Effect of pressure on the crystal structure of salicylaldoxime-I, and the structure of salicylaldoxime-II at 5.93 GPa. AB - The effect of pressure on the crystal structure of salicylaldoxime has been investigated. The ambient-pressure phase (salicylaldoxime-I) consists of pairs of molecules interacting through oximic OH...O hydrogen bonds; taken with phenolic OH...N intramolecular hydrogen bonds, these dimers form a pseudo-macrocycle bounded by an R4 4(10) motif. The dimers interact principally via pi...pi stacking contacts. Salicylaldoxime derivatives are used industrially as selective solvent extractants for copper; the selectivity reflects the compatibility of the metal ion with the pseudo-macrocycle cavity size. On increasing the pressure to 5.28 GPa the size of the cavity was found to decrease by an amount comparable to the difference in hole sizes in the structures of the Cu2+ salicylaldoximato complex and its Ni2+ equivalent. On increasing the pressure to 5.93 GPa a new polymorph, salicylaldoxime-II, was obtained in a single-crystal to single-crystal phase transition. PIXEL calculations show that the phase transition is driven in part by relief of intermolecular repulsions in the dimer-forming OH...O-bonded ring motif, and the ten-centre hydrogen-bonding ring motif of the phase I structure is replaced in phase II by a six-centre ring formed by oximic OH...N hydrogen bonds. The transition also relieves repulsions in the pi...pi stacking contacts. The intramolecular OH...N hydrogen bond of phase I is replaced in phase II by a intermolecular phenolic OH...O hydrogen bond, but the total interaction energy of the pairs of molecules connected by this new contact is very slightly repulsive because the electrostatic hydrogen-bond energy is cancelled by the repulsion term. The intra- to intermolecular hydrogen-bond conversion simply promotes efficient packing rather than contributing to the overall lattice energy. PMID- 17108666 TI - Interplay between steric and electronic factors in determining the strength of intramolecular N-H...O resonance-assisted hydrogen bonds in beta-enaminones. AB - The crystal structures of five beta-enaminones are reported: (2Z)-3-(benzylamino) 1,3-diphenyl-prop-2-en-1-one, (2Z)-3-(benzylamino)-3-(2-hydroxyphenyl)-1-phenyl prop-2-en-1-one, (2Z)-3-(benzylamino)-3-(4-methoxyphenyl)-1-(3-nitrophenyl)-prop 2-en-1-one, 2-{1-[(4-methoxyphenyl)amino]ethylidene}cyclohexene-1,3-dione and 2 {1-[(3-methoxyphenyl)amino]ethylidene}cyclohexene-1,3-dione. The structures were analysed and compared with those of similar compounds in order to establish which factors determine the range (2.53-2.72 A) of N...O hydrogen-bond distances in intramolecularly hydrogen-bonded beta-enaminones. It has been shown that, beyond electronic resonance-assisted hydrogen-bond effects modulated by substituents, the necessary requirements to produce very short N-H...O hydrogen bonding are steric intramolecular repulsions, including the embedding of an enaminonic C-C or C-N bond in an aliphatic six-membered ring. By considering the structural features it is possible to expect the strength of N-H...O hydrogen bonds adopted by specific beta-enaminones. PMID- 17108667 TI - Structures of mono-unsaturated triacylglycerols. I. The beta1 polymorph. AB - The crystal structures of the beta1 polymorphs of mono-unsaturated triacylglycerols have been solved from high-resolution laboratory and synchrotron powder diffraction data for five pure compounds, the 1,3-dimyristoyl-2 oleoylglycerol (beta1-MOM), 1,3-dipalmitoyl-2-oleoylglycerol (beta1-POP), 1,3 distearoyl-2-oleoylglycerol (beta1-SOS), 1-palmitoyl-2-oleoyl-3-stearoylglycerol (beta1-POS), 1-stearoyl-2-oleoyl-3-arachidoylglycerol (beta1-SOA) and three mixtures: the co-crystallized 1:1 molar mixture of SOS and POP [beta1-SOS/POP (1:1)] and two cocoa butters from Bahia and Ivory Coast, both in their beta-VI (=beta1) polymorph. All eight beta1 structures crystallized in the space group (P2(1)/n) and have two short cell axes (5.44-5.46 and 8.18-8.22 A), as well as a very long b axis (112-135 A). The dominant-zone problem in the indexing of the powder patterns was solved with the special brute-force indexing routine LSQDETC from the POWSIM program. Structures were solved using the direct-space parallel tempering method FOX and refined with GSAS. Along the b axis, alternations of inversion-centre-related ;three-packs' can be discerned. Each ;three-pack' has a central oleic zone, with oleic acyl chains of the molecules being packed together, that is sandwiched between two saturated-chain zones. The conformation of the triacylglycerol molecules is relatively ;flat' because the least-square planes through the saturated chains and those through the saturated parts of the olein chain are parallel. The solution of the beta1 structures is a step forward towards understanding the mechanism of fat-bloom formation in dark chocolate and has led to a reexamination of the beta2 structural model [see van Mechelen et al. (2006). Acta Cryst. B62, 1131-1138]. PMID- 17108668 TI - Structures of mono-unsaturated triacylglycerols. II. The beta2 polymorph. AB - An improved crystal structure model has been established for the beta2 polymorph of the symmetric mono-unsaturated triacylglycerol 1,3-distearoyl-2-oleoylglycerol (SOS) and the equivalent beta-V polymorph of Ivory Coast cocoa butter. In addition, the crystal structures of the beta2 polymorphs are reported for the triacylglycerols 1,3-dipalmitoyl-2-oleoylglycerol (POP) and 1-palmitoyl-2-oleoyl 3-stearoylglycerol (POS), which are, together with SOS, the major components of cocoa butter, and that of 1-stearoyl-2-oleoyl-3-arachidoylglycerol (SOA). The existence of beta2-POS and beta2-SOA has not been previously reported in the literature. All structures have been solved from high-resolution laboratory or synchrotron powder diffraction data with the direct-space parallel-tempering program FOX and refined with the Rietveld module of GSAS. All compounds crystallize in similar monoclinic unit cells (Cc) with very long b axes (>127 A). The oleic chains are packed together and sandwiched between saturated chain layers, forming acyl-chain three-packs. An analysis of the beta2 polymorphs and beta1 polymorphs [van Mechelen et al. (2006). Acta Cryst. B62, 1121-1130] shows that they contain the same three-packs and differ only in the symmetry relation between the three-packs. The three-pack build-up provides an explanation of the mechanism of the phase transition that causes the formation of fat bloom on dark chocolate. PMID- 17108669 TI - Inflammatory cell phenotyping of the pulmonary interstitium in idiopathic interstitial pneumonia. AB - BACKGROUND: Several studies have implicated the role of inflammation in the pathogenesis of lung damage in idiopathic interstitial pneumonias (IIPs). Investigations of inflammatory cells in IIP have show that eosinophils, neutrophils and T cells may be associated with a poorer prognosis. OBJECTIVES: The aim of our study was to map, by quantitative analysis, the number of inflammatory cells in the lung tissue of patients with non-specific interstitial pneumonia/non-specific interstitial pneumonia (NSIP/NSIP), acute interstitial pneumonia/diffuse alveolar damage (AIP/DAD) and idiopathic pulmonary fibrosis/usual interstitial pneumonia (IPF/UIP) and to correlate them with lung function tests and survival. METHODS: After immunohistochemical staining, we quantified the content of inflammatory cells [macrophages, neutrophils (elastase+), plasma cells, and CD3, CD4 and CD8 T lymphocytes (TLs)] in 20 NSIP, 20 DAD and 20 UIP surgical lung biopsies. RESULTS: The total density of inflammatory cells was significantly increased in DAD and NSIP when compared to UIP (p = 0.04). TLs were increased in DAD and NSIP when compared to UlP lungs (p < 0.05). The density of inflammatory cells in UIP showed significant differences in normal, intervening and dense fibrosis areas (p < 0.05). The most numerous cells infiltrating the mural fibrosis and honeycombing areas were plasma cells, neutrophils (elastase+), CD20+, CD3+, CD4+ and CD8+ (p < 0.05). In UIP, CD3+ TLs were directly correlated with forced expiratory volume in 1 s/forced vital capacity ratio x 100 (p = 0.05). CD68+ cells presented a significant positive correlation with the forced expiratory volume in 1 s (p = 0.04); neutrophil (elastase+) cells significantly correlated with residual volume (p = 0.02), residual volume/total lung capacity (p = 0.04) and carbon monoxide transfer factor (p = 0.03). The most important predictor of survival in UIP was CD3+ TLs (p = 0.05). CONCLUSION: The total density of inflammatory cells and lymphocytes presents a different distribution within the pulmonary parenchyma in AIP/DAD, NSIP/NSIP and IPF/UIP evolutionary adapted responses to injury. There is a localized distribution of inflammation in the normal, intervening and dense fibrosis areas of UIP for CD3+, associated with a lethal deterioration of the pulmonary function and poor survival. Our findings provide further evidence of the importance of inflammation in the pathophysiology of IIPs. PMID- 17108670 TI - A nicotine mouth spray for smoking cessation: a pilot study of preference, safety and efficacy. AB - BACKGROUND: Various formulations of nicotine replacement therapy are commercially available. OBJECTIVES: It was the aim of this study to test preference, safety and efficacy of a new nicotine mouth spray (1 mg/actuation; NicoNovum). METHODS: One hundred healthy smokers wanting to quit (mean age 43.1 +/- 11.2 years) were included. They were given the mouth spray, as well as 2-mg nicotine gums and nicotine oral inhalers to try out for 1 week. Subsequently, all participants were randomized into 3 groups - mouth spray (n = 50), gum (n = 25) and inhaler (n = 25) - and treated for 3 months. RESULTS: Fifty-four of the hundred smokers preferred the spray, compared with 28 and 18 who preferred the gum and inhaler, respectively (p < 0.01 for spray vs. both the gum and inhaler). At 6 months, the sustained abstinence rates were 8 (16%), 5 (20%) and 2 (8%) for the mouth spray, gum and inhaler, respectively (p values = n.s.). Adverse effects (AEs) were mostly drug-related local symptoms, most frequently reported at the end of the tryout period: out of 106 drug-related AEs, 90 were due to the spray, 11 and 5 to the inhaler and gum, respectively. The three most frequent AEs were almost exclusively caused by the spray: burning of the tongue/throat reported by 35, nausea by 18, and hiccup by 16 participants. CONCLUSIONS: Participants preferred the mouth spray over the gum and inhaler, but its use was coupled with a high rate of local AEs. The efficacy of the mouth spray seemed comparable with the one obtained by the gum and inhaler. Further studies are necessary to show whether a reduction in the dose per actuation to about 0.5 mg would result in less side effects, which in turn might yield higher long-term drug compliance and possibly higher success rates than the ones achieved with other forms of nicotine replacement therapy. PMID- 17108671 TI - Occupational immunoglobulin E-mediated asthma due to Penicillium camemberti in a dry-sausage packer. AB - A 62-year-old female sausage packer reported progressive work-related asthma, rhinitis and contact urticaria after contact to dry sausages refined by moulds. Whereas skin prick tests with commercial mould extracts were negative, the patient showed weak skin sensitization to a cultivated mould sample from a dry sausage. Specific immunoglobulin E antibodies to various moulds were demonstrated by ImmunoCAP and antibodies to the cultivated mould sample were demonstrated by enzyme allergosorbent test. The mould was identified by pheno- and genotyping as Penicillium camemberti. Five atopic controls did not show sensitization with the same tests. Crossreactivity of P. camemberti and Penicillium notatum was shown by enzyme allergosorbent inhibition tests. Although no challenge tests were considered due to the patient's airway obstruction, a diagnosis of allergic occupational asthma was made. We conclude that immunoglobulin E-mediated allergic occupational asthma due to moulds may occur in dry sausage packers. PMID- 17108672 TI - Comparative morphology of the avian cerebellum: II. Size of folia. AB - Despite the highly conserved circuitry of the cerebellum, its overall shape varies significantly among and within vertebrate classes. In birds, one of the most prominent differences among orders is the relative size of the cerebellar folia. The enlargement/reduction of individual folia is thought to relate to specific behavioral differences among taxa, but this has not been adequately tested. Here, we survey variation in cerebellar folia size among 96 species of birds and test for phylogenetic effects and correlations with behavior using a combination of conventional and phylogeny-based statistics. Overall, we found that phylogenetic history accounts for a significant amount of variation in the relative size of individual folia. Order membership, in particular, accounted for more than half of the interspecific variation in folia size. There are also complex relationships among folia such that the expansion of one folium is often accompanied by a reduction in other folia. With respect to behavioral correlates: (1) we did not find any significant correlations between folia size and reliance on trigeminal input; (2) there was some evidence supporting a correlation between strong hindlimbs and an expansion of the anterior lobe; and (3) there were significant reductions in folia I-III and expansions in folia VI and VII in species classified as strong fliers. This expansion likely reflects increased visual processing requirements in species with rapid and/or agile flight. It therefore appears that folium size is a product of both phylogenetic history and behavior in birds. PMID- 17108673 TI - On being small: brain allometry in ants. AB - Comparative neurobiologists have provided ample evidence that in vertebrates small animals have proportionally larger brains: in a double-logarithmic plot of brain weight versus body weight all data points conform quite closely to a straight line with a slope of less than one. Hence vertebrate brains scale allometrically, rather than isometrically, with body size. Here we extend the phylogenetic scope of such studies and the size range of the brains under investigation to the insects, especially ants. We show that the principle of (negative) allometry applies as well, but that ants have considerably smaller brains than any ant-sized vertebrate would have, and that this result holds even if the relatively higher exoskeleton weights of ants (as compared to endoskeleton weights of mammals) are taken into account. Finally, interspecific comparisons within one genus of ants, Cataglyphis, show that species exhibiting small colony sizes (of a few hundred individuals) have significantly smaller brains than species in which colonies are composed of several thousand individuals. PMID- 17108674 TI - Prevalence of two tumor necrosis factor gene polymorphisms in premature infants with early onset sepsis. AB - BACKGROUND: Tumor necrosis factor (TNF) is a central mediator of sepsis. The NcoI polymorphism within the TNF locus has been described as a prognostic marker for mortality in adult patients with sepsis. OBJECTIVES: The aim of our study was to investigate the genotype and allele distribution of 2 TNF gene polymorphisms in preterm infants <32 weeks of gestational age, who developed early-onset sepsis. METHODS: A double-blinded retrospective cohort study was carried out on stored Guthrie blood spot cards with group A including 67 premature infants <32 weeks of gestational age with proven early-onset sepsis and group B including 102 healthy newborn infants (>32 + 0 weeks of gestation). The genotype andallele distribution of the study population were also compared to reference groups of healthy adult volunteers (n = 252 for TNF-beta NcoI and n = 233 for -308 TNF-alpha promoter). Polymorphisms of the TNF-alpha promoter -308 region and the NcoI site of the TNF beta gene were assessed using PCR followed by melting curve analysis or NcoI digestion. The groups were compared by estimation of Hardy-Weinberg equilibrium. RESULTS: The overall allele frequency and genotype distribution of the -308 TNF alpha and NcoI polymorphism of the TNF-beta gene were comparable to the values found in the controls. CONCLUSION: The study results suggest that none of the analyzed TNF gene polymorphisms may serve as a prognostic marker for preterm infants at high risk of sepsis. PMID- 17108675 TI - Steroid-responsive recurrent encephalopathy in a patient with cerebral amyloid angiopathy. PMID- 17108676 TI - Mechanical thrombectomy in acute proximal middle cerebral artery thrombosis with the alligator retrieval device. PMID- 17108677 TI - Clinical trials and dipyridamole formulation selection. PMID- 17108679 TI - Mannheim carotid intima-media thickness consensus (2004-2006). An update on behalf of the Advisory Board of the 3rd and 4th Watching the Risk Symposium, 13th and 15th European Stroke Conferences, Mannheim, Germany, 2004, and Brussels, Belgium, 2006. AB - Intima-media thickness (IMT) is increasingly used as a surrogate end point of vascular outcomes in clinical trials aimed at determining the success of interventions that lower risk factors for atherosclerosis and associated diseases (stroke, myocardial infarction and peripheral artery diseases). The necessity to promote further criteria to distinguish early atherosclerotic plaque formation from thickening of IMT and to standardize IMT measurements is expressed through this updated consensus. Plaque is defined as a focal structure that encroaches into the arterial lumen of at least 0.5 mm or 50% of the surrounding IMT value or demonstrates a thickness >1.5 mm as measured from the media-adventitia interface to the intima-lumen interface. Standard use of IMT measurements is based on physics, technical and disease-related principles as well as agreements on how to perform, interpret and document study results. Harmonization of carotid image acquisition and analysis is needed for the comparison of the IMT results obtained from epidemiological and interventional studies around the world. The consensus concludes that there is no need to 'treat IMT values' nor to monitor IMT values in individual patients apart from exceptions named, which emphasize that inside randomized clinical trials should be performed. Although IMT has been suggested to represent an important risk marker, according to the current evidence it does not fulfill the characteristics of an accepted risk factor. Standardized methods recommended in this consensus statement will foster homogenous data collection and analysis. This will help to improve the power of randomized clinical trials incorporating IMT measurements and to facilitate the merging of large databases for meta-analyses. PMID- 17108680 TI - Changes of serum beta-endorphin by programmed exercise training are correlated with improvement of clinical symptoms and quality of life in female mitral valve prolapse syndrome. AB - BACKGROUND: Mitral valve prolapse (MVP) is a common entity in female population. Although this is a minor disease, it may cause annoying symptoms that impair quality of life (QOL), and no established therapy for this problem. The aim of this study isto examine whether programmed exercise training by treadmill in female MVP syndrome would improve clinical symptoms and QOL. METHODS: An interventional study of 39 females with MVP syndrome with treadmill exercise endurance training for 12 weeks. Every individual received training for 30 min a day, thrice a week for 12 weeks. Baseline and post-exercise at 12 weeks serum beta-endorphins were measured. Symptom improvement was assessed by the MVP symptom checklist questionnaire and the Euro-QOL-5D was used to measure QOL improvement in these females. RESULTS: The mean serum beta-endorphin increased from 0.5 to 1.68 ng/ml (p = 0.001) in the exercise group (n = 18) after 12 weeks exercise, whereas the control group (n = 21) did not show any significant changes (0.44 vs. 0.43 ng/ml). Major symptoms of MVP such as chest pain, palpitation, fatigue were improved significantly by the assessment of MVP symptom checklist. The QOL of the exercised females also showed significant changes. CONCLUSIONS: Through programmed exercise training in these MVP females, the improvement of symptoms and QOL is parallel to the increase of serum beta-endorphin. This result implicates that MVP females should initiate exercise to tackle this annoying problem. PMID- 17108682 TI - Mesenchymal stem cells in tissue engineering. AB - The repair of diseased or damaged cartilage remains one of the most challenging problems of musculoskeletal medicine. Tissue engineering advances in cartilage repair have utilized autologous and allogenic chondrocyte and cartilage grafts, biomaterial scaffolds, growth factors, stem cells, and genetic engineering. The mesenchymal stem cell has specifically attracted much attention because of its accessibility, potential for differentiation, and manipulability to modern molecular, tissue and genetic engineering techniques. Mesenchymal stem cells provide invaluable tools for the study of tissue repair when combined with a carrier vehicle/matrix scaffold, and/or bioactive growth factors. However, an underappreciated source of knowledge lies in the relationship between fetal development and adult tissue repair. The multitude of events that take place during fetal development which lead from stem cell to functional tissue are poorly understood. A more thorough understanding of the events of development as they pertain to cartilage organogenesis may help elucidate some of the unknowns of adult tissue repair. PMID- 17108683 TI - Characterization of ectomesenchymal cells isolated from the first branchial arch during multilineage differentiation. AB - Ectomesenchymal cells isolated from the first branchial arch have the potential to differentiate into a variety of cell lineages both in vitro and in vivo. This study was aimed to confirm the plasticity of multilineage differentiation with molecular and cellular characterization. Monolayer cultures of ectomesenchymal cells harvested from the first branchial arch primordia in embryonic day 9.5 BALB/c mice were passaged 3 times before analysis. Staining with antibodies against S-100, p75 and vimentin suggested that the population of stem cells originated from ectomesenchyme, with few contaminating cells stained for cytokeratin. Then, cells were transferred to adipogenic, osteogenic, chondrogenic and odontogenic media. The initiation of controlled differentiation was determined with histological assays, and the expression of tissue-specific genes was detected using immunocytochemical staining and reverse transcription polymerase chain reaction. The adipogenic ectomesenchymal cells showed accumulation of lipid vacuoles and expression of lipoprotein lipase and peroxisome proliferator-activated receptor gamma(2). Following osteoinduction, the fibroblast-like cells became cuboidal and formed mineralized nodules. In addition, the expression of mRNA encoding osteocalcin and osteopontin proved osteogenesis at the molecular level. Chondrogenic lineage expressed collagen type II, aggrecan and Sox9 with a low level of collagen type I in monolayer culture. Odontogenesis was determined by dentin sialophosphoprotein, collagen type I and dentin matrix protein 1 expression. Therefore, we have demonstrated that ectomesenchymal cells from the first branchial arch are capable of extensive multilineage differentiation in vitro, controllable by the culture environment. This makes them a relevant and valuable source of stem cells for research of craniofacial development and tissue engineering of restoration. PMID- 17108684 TI - Adipose tissue engineering by human adipose-derived stromal cells. AB - Tissue engineering has emerged as a promising alternative approach to current clinical treatments for restoration of soft tissue defects. The purpose of this study was to investigate adipose tissue formation in vitro and in vivo by using human adipose-derived stromal cells (ADSCs) utilizing a gelatin sponge (Gelform) as a scaffold. Adipogenic potentials of human ADSCs were demonstrated by Oil-O red staining and cellular morphology. After seeding human ADSCs in a density of 3 x 10(6) cells/ml on three-dimensional gelatin sponges, tissue-engineered constructs were exposed to adipogenic differentiation medium for in vitro studies and implanted in the backs of severe combined immunodeficient (SCID) mice for in vivo adipose regeneration. Adipogenesis of ADSC-seeded gelatin sponges was confirmed by Oil-O-red staining after 4 weeks of in vitro incubation. The optical density of the elution from Oil-O-red staining of adipogenic constructs is significantly higher than that of the control group (p < 0.05, n = 4). With short term in vitro differentiation, adipogenic constructs turned into fat tissue 4 weeks after in vivo implantation, confirmed by biochemical and immunohistochemical examination. No adipogenic-morphological change or fat formation was observed in in vitro or in vivo studies when ADSCs were exposed to a control medium without adipogenic stimulation. These results indicate that engineered adipose tissue can be achieved using human ADSCs and biocompatible and degradable gelatin sponges. PMID- 17108685 TI - Structure of the midterm placenta of the spotted hyena, Crocuta crocuta, with emphasis on the diverse hemophagous regions. AB - The hyena placenta is unique among carnivores in being hemochorial. It also has areas of erythrocyte uptake that differ from those seen in more commonly studied carnivores. The availability of timed midterm pregnancies made it possible to examine the organization of the placenta, the distribution of regions of columnar trophoblast and the nature of the heterophagous and hemophagous regions in well preserved material. The labyrinth of the placenta is orderly arranged with periodic primary villi that have a surface of syncytial trophoblast. Secondary projections from the primary villi anastomose extensively. The junctional extreme of each primary villus forms an expanded tip covered by columnar trophoblast cells. Marginal to the hemochorial placenta, the paraplacenta is exposed to varying amounts of extravasated maternal blood which the cytotrophoblast cells ingest and destroy. Columnar trophoblast cells at the tips of the primary villi, in the villi of paraplacental margins of the labyrinth, in isolated patches within the labyrinth and beneath the allantochorionic plate are all capable of and are variably involved in erythrophagocytosis. Variations in the structure and the lectin histochemistry of the columnar trophoblast cells indicate variation depending on the exposure of these cells to erythrocytes and/or other histotrophic materials. The widespread distribution of the hemophagous regions suggests a nutritive function in addition to an iron transfer function of the columnar cytotrophoblast. PMID- 17108686 TI - Direct magnetic tubular cell seeding: a novel approach for vascular tissue engineering. AB - Optimizing seeding efficiency, reducing delayed culture periods and mimicking native tissue architecture are crucial requirements for the development of seeding procedures in tissue engineering. In vascular applications, the tubular geometry of the grafts further hampers the efficient delivery of cells onto the scaffold. To overcome these limitations, a novel technology based upon the use of magnetic fields is presented in this study: a radial magnetic force drives the cells immediately onto the luminal surface of a tubular scaffold and immobilizes the cells on the substrate's surface promoting cell attachment. Human smooth muscle cells (SMCs) labeled with CD44 magnetic Dynabeads were successively seeded onto the luminal surface of a tubular shaped collagen membrane. After 5 h, one additional layer of human umbilical vein endothelial cells (HUVECs) labeled with CD31 magnetic Dynabeads was seeded onto the luminal SMCs. The co-culture was incubated during 5 days prior to analysis. Cell viability and expression profiles were preserved during the entire seeding process. Histological examination of the constructs highlighted densely packed multilayers of SMCs covered by a monolayer of endothelial cells. SEM inspection confirmed a heterotypic multilayer assembly formed by multiple layers of elongated SMCs covered by a single layer of endothelial cells. Seeding kinetics of HUVECs and SMCs showed over 90% seeding efficiency after 20 and 40 min magnetic exposure respectively. Magnetically induced cell seeding provides a valuable tool for rapid seeding procedures of tubular scaffolds while complying with the histological architecture of tissue. PMID- 17108687 TI - Apolipoprotein E epsilon4 allele frequency and age at onset of Alzheimer's disease. AB - The age distribution of the epsilon4 allelic form of the apolipoprotein E gene (APOE) was investigated in 630 patients with Alzheimer's disease (AD) with onset age ranging from 35 to 90 years. Overall, mean age at onset in APOE epsilon4 allele bearers was significantly later than that in nonbearers. However, when stratified into early onset AD (EOAD) and late onset (LOAD) groups, mean age at onset in EOAD cases bearing APOE epsilon4 allele was later than that in those EOAD cases without epsilon4 allele, whereas in LOAD mean age at onset in cases bearing APOE epsilon4 allele was earlier than in those without epsilon4 allele. When analysed by decade, it was observed that 37% of the total number of APOE epsilon4 allele bearers, and 43% of total number of cases with APOE epsilon4/epsilon4 genotype fell into the 60-69 years age class. Hence, APOE epsilon4 allele frequency, at 0.44, was highest in the 60-69 years age class, progressively decreasing either side of this age group. APOE epsilon4 allele therefore has its maximum impact between onset ages of between 60 and 70 years. PMID- 17108688 TI - Alterations in the expression of ATP-sensitive potassium channel subunit mRNA after acute peripheral nerve and spinal cord injury. AB - ATP-sensitive potassium channels (K(ATP)) are involved in the regulation of potassium homeostasis in the nervous system, and they may play an important role in acute peripheral nerve and spinal cord injury. Here, the expression of the K(ATP) genes was monitored by reverse transcription polymerase chain reaction (RT PCR) in the rat dorsal root ganglion, spinal cord and cerebral cortex following acute sciatic nerve and spinal cord injury. Electrophoresis of the RT-PCR products showed that in comparison with the normal rats, the K(ATP) mRNA expression level was up-regulated for the Kir6.2 subunit in the rat dorsal root ganglion 4 and 24 h after the acute sciatic nerve injury (142.7 +/- 23.0 and 135.5 +/- 21.0%, p < 0.05, vs. control, n = 3), and both Kir6.1 and sulphonylurea receptor 2 mRNA were increased in the spinal cord during the same time period after the acute spinal cord injury (266.5 +/- 67.1 and 248.7 +/- 67.7%; 145.1 +/- 42.6 and 152.6 +/- 44.3%, p < 0.05, vs. control, n = 3). No significant changes of K(ATP) genes were observed in the cerebral cortex among both sciatic-nerve- and spinal-cord-injured animals. These results suggest that acute peripheral nerve and spinal cord injury provoke different regulations of K(ATP) gene expression in the peripheral and central nervous system. PMID- 17108689 TI - Ca channel currents inhibited by serum from select patients with Guillain-Barre syndrome. AB - We performed an electrophysiological study demonstrating inhibition of spontaneous muscle action potentials within a coculture of rat muscle and spinal cord by exposure to serum, as well as purified IgG, from patients with the acute motor axonal neuropathy (AMAN) variant of Guillain-Barre syndrome (GBS). However, exposure to serum from two patients with the acute inflammatory demyelinating polyneuropathy (AIDP) form of GBS had no effect. Using a whole-cell recording technique, we then investigated the effects of serum and purified IgG from patients with GBS on voltage-dependent calcium channel (VDCC) currents in nerve growth factor-differentiated PC12 cells. Serum from patients with GBS (AMAN) inhibited VDCC currents in PC12 cells, which was fully reversible by washing with the bath solution. Similarly, purified IgG from the serum of two patients with GBS (AMAN) also inhibited VDCC currents in PC12 cells. In contrast, sera from patients with AIDP and healthy volunteers did not affect VDCC currents in PC12 cells. These results suggest that muscle weakness in some patients with GBS might be induced by inhibition of Ca2+ channel currents within motor nerve terminals. PMID- 17108690 TI - Histological and immunohistological changes of the skeletal muscles in older SJL/J mice. AB - SJL/J mice have been studied as the model animals for autoimmunological diseases. Recently it was clarified that SJL/J mice have a defect of dysferlin. Human limb girdle muscular dystrophy 2B and Miyoshi myopathy also have a defect of dysferlin. In this study we present the histological and immunohistological changes in the natural course. Histological study revealed that SJL/J mice had inflammatory, degenerative changes, and neurogenic changes in later ages. As for interstitial inflammatory cells, the macrophages were dominant in any age, and in the T cell subset, the CD4+ T cells were more abundant than the CD8+ T cells, and few B cells were seen. The laboratory data showed a high level of creatine kinase in all ages. It is suspected that the inflammatory changes were induced by the primary immunological abnormality or by the defect of dysferlin in SJL/J mice. PMID- 17108691 TI - Monozygotic twins suffering from Huntington's disease show different cognitive and behavioural symptoms. AB - Monozygotic male twins, carrying the same number of trinucleotide repeats in the IT 15 Huntington disease (HD) gene, showed a different clinical course. Patient 1 presented with anxiety and chorea at the age of 40. Patient 2 showed persecution paranoia and motor impersistence at the age of 42. Both patients were monitored for 30 months using currently recommended motor and behaviour scales. No differences were observed in motor scoring besides small interevaluation fluctuations. However, on the cognitive and behaviour scales, patient 1 showed a significant worsening when compared with patient 2. Our cases support the belief that the motor symptoms and signs in HD are highly dependent on the trinucleotide expansion. However, the differences in the evolution of mental status in our patients suggest that other still unknown environmental factors are important in the phenotypic expression of Huntington's disease. PMID- 17108692 TI - Restless legs syndrome in hereditary spastic paraparesis. AB - This study was designed to investigate the prevalence and determinants of the association of restless legs syndrome (RLS) and hereditary spastic paraparesis (HSP). Therefore, 132 patients with HSP were evaluated concerning the symptoms of RLS by a standardized questionnaire. RLS was supposed when patients met all of the established four essential criteria of RLS defined by the International RLS Study Group. In addition, we studied the relationship between RLS and age at HSP symptom onset and evaluated the severity of RLS symptoms. Out of 59 responses, RLS was found in 27 HSP patients (15 male, 12 female) so that RLS was more frequent in the total HSP group (20.5%; 27/132) than in previous population-based studies (about 11%). In all 27 patients, the diagnosis of RLS was established based on an additional personal interview. The probability to develop RLS did not increase with higher age. Age at onset of HSP symptoms in the HSP group with RLS (27.7 +/- 12.6 years) and the HSP group without RLS (37.0 +/- 16.9 years) differed significantly (p = 0.04). Most of the patients with RLS showed a moderate and severe grade on the RLS severity score. Only 8 patients had previously been diagnosed to have RLS and were on medication. The data of this screening for RLS provided evidence that patients with HSP are particularly susceptible to develop RLS. Consequently, special emphasis should be put on the diagnosis criteria of RLS in HSP patients. PMID- 17108693 TI - Two patients with tremor caused by cortical lesions. AB - Myoclonic tremor due to cortical lesions is a rare condition and must be distinguished from other causes of tremor. This is because the treatment strategies of tremor may differ due to the various etiologies. We present here two cases with myoclonic tremor caused by parietal cortical lesions showing tremulous finger movement provoked by action and posture. Clinical and electrophysiological features of the patients were reported and compared with the features of the patients with cortical tremor in association with cortical reflex myoclonus. Both of our patients responded well to anticonvulsants such as valproate and clonazepam. In patients with acute postural tremor, cortical lesions such as mass occupying lesions, ischemic lesions and arteriovenous malformations should be taken into consideration. PMID- 17108694 TI - Executive performance on the Wisconsin Card Sorting Test in mesial temporal lobe epilepsy. AB - Executive dysfunction assessed by the Wisconsin Card Sorting Test has been observed in patients with mesial temporal lobe epilepsy (TLE); however, the mechanism of executive impairment is unclear. We therefore investigated the potential contribution of the hippocampus toward executive dysfunction in 85 mesial TLE patients, and in a control group of 34 patients with neocortical TLE. Standardized regression-based methodology was used for correcting test-retest bias. We found that 56% of mesial TLE patients had impaired sorting performance, with 30% showing severe impairment. A lower full-scale intelligence quotient, older age, longer duration, and preoperative use of topiramate were significantly correlated with poorer preoperative sorting abilities. However, neither these variables nor postoperative discontinuation/reduction of topiramate were related to postoperative changes in sorting abilities. Only preoperative sorting abilities were negatively correlated with postoperative changes in executive performance. That is, patients with better preoperative executive ability showed greater postoperative deterioration in executive function, whereas those with poorer preoperative sorting ability had greater improvement in executive function after mesial temporal resection. There were no differences in card sorting ability between the mesial and neocortical TLE groups. In conclusion, our results suggest that impairment of card sorting performance in the mesial TLE group cannot be explained only by hippocampal dysfunction, but that other processes (possibly epileptic discharges propagated to the extratemporal area) must play a predominant role. PMID- 17108695 TI - Ictal spitting in a patient with dominant temporal lobe epilepsy: Supporting evidence of ictal spitting from the nondominant hemisphere. PMID- 17108696 TI - Patterns of oral-nasal balance in normal speakers with and without cleft palate. AB - OBJECTIVE: The aero-acoustic aspects of nasalization that distinguish speakers with velopharyngeal adequacy from those with velopharyngeal inadequacy are well known. Differences in aerodynamic aspects of nasalization also have been demonstrated in normal speakers with cleft palate and their noncleft counterparts. The purpose of this study was to compare the amplitude and temporal patterns of oral-nasal balance in speakers with cleft palate and normal speech to those with normal speech without cleft palate. PATIENTS AND METHODS: Using the Nasal Accelerometric Vibrational Index technique, the time course and amplitude of oral-nasal balance were studied in 10 children with cleft palate and velopharyngeal adequacy and 10 noncleft controls. The speech sample included syllable, word, and sentence contexts, most of which were devoid of nasal consonants. RESULTS: The results indicate few differences between the two groups in contexts devoid of nasal consonants. However, significant differences were evident with regard to temporal aspects of oral-nasal balance in a nasal-oral consonant word context. CONCLUSIONS: Subtle structural-motor differences that may exist between normal speakers with and without cleft palate may be more evident when the spatiotemporal requirements for velar movement become more complex and result in a greater possible combination of oral-nasal impedance ratios. PMID- 17108697 TI - Laryngeal microweb and vocal nodules. Clinical study in a Brazilian population. AB - The etiology and pathogenesis of laryngeal microwebs are heterogeneous, and in most cases they are an incidental finding. It has also been suggested that microwebs could be a familial trait, representing a postblastogenic embryonic vestigial structure that might alter the biomechanical and vibratory properties of the vocal fold. Vocal nodules are small benign swellings along the margins of the vocal cords, with preferential location at the junction of the anterior and middle thirds, and usually resulting from mechanical trauma. The authors studied a sample of 107 patients with vocal nodules, looking for a possible correlation with microwebs due to the predicted involvement in the vibration of vocal cord margins. Glottic proportion, type of glottic closure, nodule location and the main complaints in patients with microwebs were compared with those in a sample of patients without microwebs. In the present study, microwebs were found in 9.4% of the patients, who showed much smaller glottic proportion than patients with micronodules only. Furthermore, vocal nodule location was not related to the presence and/or absence of laryngeal microwebs. PMID- 17108698 TI - Phonological profile of Spanish-Catalan children with specific language impairment at age 4: are there any changes over time? AB - The phonology of a group of Spanish-Catalan children with specific language impairment (SLI, n = 5), who had been analysed at age 3;10, is now analysed at age 4;09 and compared with two control groups: an age-matched control (n = 5) and a language level control (measured using the mean length of utterance by words; n = 5). The children with SLI continue to show a delay in the acquisition of segments, syllabic structures and in the use of the simplification processes, but not in word structures, compared with their age-matched controls. Children with SLI also display significant differences compared with their language level controls, but not in the same areas as observed at age 3: the differences are now in nasals and liquids at the segmental level, and in CCV, CVC and other complex structures at the syllabic level. There are also some simplification processes that seem to be more prevalent in these children than in their language level controls: absence of trill, cluster reductions and consonant deletions. The results enable us to interpret SLI as more than a delayed development and to show the differences in the profiles over time. PMID- 17108699 TI - [Analysis of voice parameters after conservative treatment of laryngeal paralysis with conventional voice exercises or neuromuscular electrophonatory stimulation]. AB - Two groups of patients with unilateral or bilateral paresis of the nervus laryngeus inferior were treated with conventional voice therapy or neuromuscular electrophonatory stimulation (NMEPS). A few had additional paresis of the nervus laryngeus superior. After therapy pareses resolved or were compensated for. Successful therapy outcome can be verified on the basis of voice quality parameters. Significant improvements for both therapy groups were observed in case of parameters indicating frequency irregularities and noise-to-harmonic ratio. Only the patients treated with NMEPS reached significantly better results in case of soft phonation index after resolution of paresis. PMID- 17108700 TI - Preliminary findings on the classical singer's use of the pectoralis major muscle. AB - OBJECTIVE: This study aims to further characterize the muscle activity that influences the posture and breathing utilized for classical singing. The activity in the pectoralis major (PC) muscle and the phasing of PC activity to sternocleidomastoideus (STM) activity and upper thorax (UTX) movement were investigated. MATERIAL AND METHODS: Seven professional classical singers (3 sopranos, 1 mezzo, 1 tenor and 2 baritones) and 8 advanced classical singing students (4 sopranos, 1 mezzo, 1 tenor and 2 baritones) participated. Electromyographic activity was recorded from the PC and STM muscles on the right side. UTX movement was traced with a strain gauge sensor placed around the upper thorax. Different arias, freely chosen by the singers from their professional repertoire, served as singing tasks. All subjects performed their task 3 times with variation in vocal loudness (normal, forte, piano). RESULTS: It was observed that a majority of the singers activated the PC during inhalation and that > 50% of the singers activated it during parts of phonation. In general, however, the activity of the PC was relatively low during phonation. CONCLUSION: This study shows that the PC, in idiosyncratic patterns, could be involved in the inspiratory effort recruited when positioning the UTX during inhalation and phonation. PMID- 17108701 TI - Effectiveness and efficiency of a universal newborn hearing screening in Germany. AB - The decision to mandate, finance, and implement a universal newborn hearing screening (UNHS) requires the evaluation of its therapy-directed benefit by comparing (1) a procedure employing a UNHS with (2) a targeted screening for at risk babies for neonatal hearing disorders and (3) a procedure without systematic screening. In a cohort study the outcome of the UNHS program of Hessen in 2005 with 17,439 screened newborns was analyzed. Validity, effectiveness, and efficiency were evaluated and compared to a sample of 98 Hessian and 355 German children who were detected in 2005 as hearing-impaired but not by an UNHS. The UNHS group had a PASS rate of 97.0%. Forty-nine hearing-impaired children were diagnosed at a median age of 3.1 months and treated at a median age of 3.5 months. Corresponding values for the Hessian non-UNHS group were 17.8 and 21.0 months. For Germany the median age at diagnosis was 39.0 months. The age at therapy onset correlated negatively with parameters of speech/language and psychosocial development. A targeted screening would have resulted in a low sensitivity of 65.3%. Hence, a UNHS is the most effective way to an early therapy of neonatal hearing disorders with an optimal outcome. PMID- 17108702 TI - Association of beta-defensin 1 single nucleotide polymorphisms with atopic dermatitis. AB - BACKGROUND: Atopic dermatitis (AD) is a chronic multifactorial allergic disease with unclear etiology. The antimicrobial human beta-defensin 1 is chemotactic for dendritic cells, which are important regulators of allergic immune responses. In an attempt to identify useful markers that could predict susceptibility to AD, we investigated single nucleotide polymorphisms (SNPs) of the beta-defensin 1 gene (DEFB1) with potential functional consequences. METHODS: Four SNPs of the DEFB1 gene were genotyped either by real-time polymerase chain reaction or polymerase chain reaction-restriction fragment length polymorphisms in 59 patients with AD and 151 controls from the Mexican population. Correlation analyses were carried out between genetic, environmental and clinical variables in AD patients. RESULTS: The genotypes associated with susceptibility to AD and no other allergy were 692 GG (OR = 3.21, 95% CI 1.37-7.34) and 1654 AA (OR = 17.37, 95% CI 1.62 860.83). The allele 668 C is a risk factor for AD (OR = 2.23, 95% CI 1.22-4.01) and the allele A in site 1836 correlates with earlier age at onset (Spearman's rho = 0.232; p = 0.03). The prolonged duration of breastfeeding correlates with earlier age at onset as well as with the severity of AD. CONCLUSIONS: The DEFB1 gene is probably involved in the incidence and development of AD, but additional functional studies will be necessary to understand the biological role of these SNPs. PMID- 17108703 TI - Dexamethasone alters bronchoalveolar lavage fluid proteome in a mouse asthma model. AB - BACKGROUND: Glucocorticoid is the most effective anti-inflammatory agent for asthma. The spectrum of protein targets that can be regulated by glucocorticoid in asthma is not fully understood. The present study tried to identify novel protein targets of dexamethasone in allergic airway inflammation by analyzing the proteome of mouse bronchoalveolar lavage (BAL) fluid. METHODS: BALB/c mice sensitized and challenged with ovalbumin (OVA) showed increased pulmonary inflammatory cell infiltration, airway mucus production and serum OVA-specific IgE level. Dexamethasone inhibited all these allergic airway inflammation endpoints. BAL fluid proteins were resolved by two-dimensional gel electrophoresis and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. RESULTS: The levels of 26 BAL fluid proteins were found to be markedly altered by dexamethasone. A family of chitinases (Ym1, Ym2 and acidic mammalian chitinase, AMCase), lungkine, gob-5, surfactant protein D and polymeric immunoglobulin receptor have been found for the first time to be downregulated by dexamethasone in allergic airways. The downregulatory effects were confirmed by immunoblotting and RT-PCR analyses. Dexamethasone was also shown to significantly inhibit lavage fluid chitinase bioactivity. In addition, dexamethasone promoted airway expression of vitamin D-binding protein, heptoglobin and alpha(1)-antitrypsin. CONCLUSIONS: Among all these newly identified protein targets of dexamethasone, AMCase and gob-5 have been shown to be pro-inflammatory in asthma. Downregulation of AMCase and gob-5 may be considered as two novel anti-inflammatory actions of glucocorticoid in asthma. PMID- 17108704 TI - Reference values of total serum IgE and their significance in the diagnosis of allergy in young European adults. AB - Allergic sensitization mediated by immunoglobulin E (IgE) is the basis of allergic diseases, and elevated total IgE, in spite of some well-known limitations, is frequently included as a diagnostic criterion for allergic diseases. The reference value of total IgE (IgE-t) in the literature (1.5-144 kU/l) was established almost 2 decades ago. The aim of this study was to establish IgE-t reference values, establishing an updated cutoff value able to identify atopic subjects, defined as a positive CAP-radioallergosorbent test to at least one of a panel of common allergens, among young European adults. The study included 6,670 subjects from 10 Western European countries within the framework of the European Community Respiratory Health Survey II. IgE-t and specific IgE (IgE-s) were measured for the main inhalant allergens; IgE-s in class 0 for all allergens (66.2%) characterized non-atopy. The reference values were estimated by means of linear regression using a 50% random subsample of non atopic subjects. Two non-atopic subsamples were examined so that one subsample could be used to establish reference IgE-t values, and these values were compared to those in the second non-atopic subsample to validate the findings. Sensitivity and specificity for atopy were assessed on the other 50% of non-atopic and on all atopic subjects. The 95th percentile of IgE-t reference values in non-smokers was 148 kU/l in women and 169 kU/l in men, while it was 194 and 220 kU/l in female and male smokers, respectively: serum IgE-t above the 95th percentile identifies <32% and above the 99th percentile <20% of atopic adults (low sensitivity), but a serum IgE-t below the 95th percentile identifies >90% and below the 99th percentile identifies >95% of non-atopic adults (good specificity). Due to the adequate specificity, IgE-t values exceeding the normal limits confirm a suspected atopic status; however, because of the low sensitivity, values below the cutoff seem not to exclude an atopic status with sufficient accuracy. PMID- 17108705 TI - Mechanisms of vasculitis: how pauci-immune is ANCA-associated renal vasculitis? AB - Both the innate and the acquired immune system are involved in the pathophysiology of renal vasculitis. However, anti-neutrophil cytoplasmic antibody (ANCA)-associated renal vasculitis is characterized by a 'pauci-immune' pattern of immunofluorescence during kidney biopsy, indicating the relative lack of immunoglobulin and complement deposition within the kidney. On the other hand, evidence is accumulating that ANCA, autoantibodies against constituents of primary granules of neutrophils and the lysosomes of monocytes, play a pathogenic role in renal vasculitis. In this review we will discuss both in vitro and in vivo experimental data providing compelling evidence that ANCA are a primary pathogenic factor in renal vasculitis, mainly by augmenting leukocyte-endothelial interactions. We will also address novel data, pointing at the role of, in addition to ANCA, non-specific proinflammatory signals. Finally, we propose a working hypothesis of the pathogenesis of ANCA-associated renal vasculitis. PMID- 17108706 TI - Rethinking the triggering inflammatory processes of chronic periaortitis. AB - Chronic periaortitis (CP) is an uncommon inflammatory disease which primarily involves the infrarenal portion of the abdominal aorta. However, CP should be regarded as a generalized disease with three different pathophysiological entities, namely idiopathic retroperitoneal fibrosis (RPF), inflammatory abdominal aortic aneurysm and perianeurysmal RPF. These entities share similar histopathological characteristics and finally will lead to fibrosis of the retroperitoneal space. Beside fibrosis, an infiltrate with variable chronic inflammatory cell is present. The majority of these cells are lymphocytes and macrophages as well as vascular endothelial cells, most of which are HLA-DR positive. B and T cells are present with a majority of T cells of the T-helper phenotype. Cytokine gene expression analysis shows the presence of interleukin (IL)-1alpha, IL-2, IL-4, interferon-gamma and IL-2 receptors. Adhesion molecules such as E-selectin, intercellular adhesion molecule-1 and the vascular cell adhesion molecule-1 were also found in aortic tissue, and may play a significant role in CP pathophysiology. Although CP pathogenesis remains unknown, an exaggerated inflammatory response to advanced atherosclerosis (ATS) has been postulated to be the main process. Autoimmunity has also been proposed as a contributing factor based on immunohistochemical studies. The suspected allergen may be a component of ceroid, which is elaborated within the atheroma. We review the pathogenesis and the pathophysiology of CP, and its potential links with ATS. Clinically relevant issues are summarized in each section with regard to the current working hypothesis of this complex inflammatory disease. PMID- 17108707 TI - Dialysis membrane-induced oxidative stress: role of heme oxygenase-1. AB - Dialysis membranes have been reported to induce monocyte apoptosis. We studied the role of oxidative stress in the induction of dialysis membrane-induced monocyte apoptosis. Superoxide dismutase, a superoxide scavenger, prevented dialysis membrane-induced monocyte apoptosis. Similarly, other antioxidants also inhibited dialysis membrane- induced apoptosis. In addition, the interaction of dialysis membranes with monocytes was associated with the generation of molecules leading to oxidative stress such as superoxide and TBARS. Interestingly, pre induction of heme oxygenase (HO)-1 by hemin prevented dialysis membrane-induced monocyte apoptosis, whereas inhibition of HO-1 activity (treatment with tin protoporphyrin, SN-P) enhanced dialysis membrane-induced monocyte apoptosis. We suggest that oxidative injury associated with dialysis membrane and monocyte interaction plays a role in monocyte injury. Pre-induction of HO-1 may attenuate dialysis membrane-induced monocyte apoptosis. PMID- 17108708 TI - Extent of glomerular tubularization is an indicator of the severity of experimental acute kidney injury in mice. AB - BACKGROUND: Acute kidney injury (AKI) secondary to ischemia continues to be a major clinical problem due to its high morbidity and mortality, and limited treatment options. Animal models are critical to both the study of the pathophysiology of AKI and the development of new interventions. Although histological changes at the glomerulotubular junction have been described in AKI, we examined here whether the extent of glomerular tubularization correlates with the degree of renal insufficiency in this condition. METHODS: Groups of mice with ischemia/reperfusion AKI were utilized in which the severity of renal insufficiency was defined. The resulting level of glomerular tubularization was analyzed, and the involved cell type was identified by immunohistochemistry and electron microscopy. RESULTS: The extent of glomerular tubularization increased significantly with the degree of renal insufficiency. Low level glomerular tubularization was present in normal mouse kidneys, while it was more common and increasingly circumferential in mice with more severe loss of kidney function. The parietal monolayer of cuboidal cells in glomeruli was contiguous with proximal tubular cells, showing a well-developed luminal brush border and positive staining for proliferating cell nuclear antigen and Lotus tetragonolobus, a proximal tubular cell-specific lectin. CONCLUSION: Increased levels of glomerular tubularization represent a poorly understood response to ischemic AKI in mice. As such, this glomerular 'tubularization score' may be useful to complement standard injury scores in experimental and, if detected, in clinical AKI. PMID- 17108709 TI - Potential clinical targets of repetitive transcranial magnetic stimulation treatment in schizophrenia. AB - Despite the introduction of atypical antipsychotic drugs, treatment-resistant symptoms still represent a serious problem in schizophrenia. Currently, there is evidence from clinical studies suggesting that treatment with repetitive transcranial magnetic stimulation (rTMS) may improve schizophrenia symptoms. Our review provides an overview of clinical rTMS studies in schizophrenic patients. A systematic search of the literature (Cochrane and Medline databases up to December 2005) was conducted. Most studies showed methodological problems due to their explorative character and small sample sizes. In some studies, a treatment effect of high-frequency rTMS applied over the prefrontal cortex was seen with respect to negative symptoms. On the other hand, low-frequency rTMS in the temporal lobe area might lead to a suppression of auditory hallucinations. It is concluded that larger sham-controlled studies are required to allow an adequate assessment of the clinical and neurobiological effects of rTMS in schizophrenic patients. The currently available data provide insufficient evidence to support the use of rTMS as an adjuvant treatment for schizophrenic psychopathology, but encourage further investigation of rTMS as a novel treatment approach. PMID- 17108710 TI - Social factors but not stress-coping styles predict relapse in detoxified alcoholics. AB - Alcohol-dependent patients face a substantial risk of relapse after detoxification. Though psychosocial stress and coping strategies are regarded as major contributing factors in returning to drinking, the direct effects of coping styles on relapse are not clear. In this treatment outcome study, a mixed gender sample of 130 detoxified and well-characterized alcohol-dependent patients (37 women) was followed up over a period of 12 months after 6 weeks of inpatient treatment. Patients had completed a comprehensive baseline assessment, including a stress coping questionnaire (SVF120). We hypothesized that these individual stress coping styles would contribute to treatment outcome. A logistic regression analysis was used to evaluate the impact of stress coping styles, as well as the effect of pretreatment drinking and social characteristics on relapse. Approximately half the patients (49%) relapsed within 1 year after treatment. In contrast to our hypothesis, stress coping styles did not predict relapse. However, significant predictors of relapse were social factors related to living situation (living alone), marital status (being separated from the spouse) and pretreatment frequency of alcohol intake. These findings suggest that a partnership is more relevant for the risk of relapse than stress coping styles. PMID- 17108711 TI - Diffusion tensor imaging of the corpus callosum in addiction. AB - Diffusion tensor imaging, a novel technique with an increased capability of detecting abnormalities in the white matter, has increasingly been employed in the study of the biology of addictions. A comprehensive search from a range of databases was conducted and publications on this topic were selected. Nine reports, eight published and one unpublished, met criteria for inclusion, five on alcoholism, three on cannabis and one on cocaine use. Findings of this review suggest focal disruption of commissural connectivity in the corpus callosum. In alcoholism, the genu and splenium were particularly affected with a different pattern in men and women, and an association with age and duration of substance use. In cocaine dependence, the genu and rostral body showed significant damage. Cannabis consumption may be associated with white matter disruption, but there is not sufficient evidence to support pathological changes in the corpus callosum. The improved detection of white matter pathology with diffusion tensor imaging supports the importance of future research in this field. PMID- 17108712 TI - Reduced tumor growth and angiogenesis in endoglin-haploinsufficient mice. AB - Endoglin is a transforming growth factor-beta(1) (TGF-beta(1)) accessory receptor which is highly expressed in tumor vessels. To study the role of endoglin in tumor growth and angiogenesis we induced a highly vascularized tumor in mice heterozygous for endoglin (Eng+/-) and in their control littermates (Eng+/+) by injecting 10(6) Lewis lung carcinoma (3LL) cells subcutaneously. Nine days after injection, the tumor was removed and weighed. Capillary density (CD31 immunohistochemistry), hemoglobin content and vascular cell adhesion molecule-1 (VCAM-1) expression were used to assess tumor vascularization. Tumor perfusion rate was measured by laser-Doppler technique. Expression of the hypoxia-inducible factor (HIF), endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) were determined by Western blot analysis. The aerobic metabolism and oxygen dependency were inferred from the measurement of ATP in tumoral tissue. Tumor weight, capillary density, hemoglobin and VCAM-1 were reduced by about 30% in Eng+/- compared to Eng+/+ littermates. The protein levels of eNOS and phosphorylated eNOS were significantly reduced in Eng+/- compared to Eng+/+ mice. HIF expression was slightly reduced whereas VEGF level was slightly increased in Eng+/- compared to Eng+/+. Tumor tissue levels of ATP and ADP were similar in both types of mice. These data demonstrate that endoglin plays a major role in tumor neoangiogenesis. PMID- 17108713 TI - Overexpression of PTEN suppresses growth and induces apoptosis by inhibiting the expression of survivin in bladder cancer cells. AB - The tumor suppressor gene PTEN, which encodes a multifunctional phosphatase protein, is mutated in a variety of human cancers. Several reports have indicated that it has growth-suppressive and proapoptosis properties and displayed an altered expression pattern during human oncogenesis. Overexpression of PTEN leads to decreasing cell growth and tumorigenicity in vitro and in vivo. In the present study, we further demonstrated that overexpression of PTEN mediated by adenovirus suppressed bladder cancer cell growth and significantly induced apoptosis, through downregulating of survivin and activating of caspase cascades. Our results indicate that Ad-PTEN exerts its tumor suppressive effect on bladder cancer cells through inhibiting survivin and upregulating caspase-related proteins. Thus Ad-PTEN may be potentially therapeutic for the treatment of bladder cancers. PMID- 17108714 TI - [An advanced pulmonary adenocarcinoma case in which TS-1 proved effective]. AB - A sixty-two-year-old female presented with non-small-cell lung carcinoma with multiple bone metastasis. Combination chemotherapy with CBDCA and GEM was implemented, but as no effect was obtained, TS-1 was begun with a single course of oral administration of 100 mg/day continuously for 28 days followed by a 14 day rest. Following the start of TS-1, PR of the primary tumor was obtained, and whole-body bone scintigraphy revealed a reduction in abnormal areas of uptake. The woman continues to receive TS-1 orally as an outpatient. PMID- 17108715 TI - [A case of advanced gastric cancer with malignant ascites responding to weekly paclitaxel therapy]. AB - A 54-year-old woman with scirrhous gastric cancer in the upper third area was admitted to our hospital. She was diagnosed with advanced gastric cancer that was inoperable due to peritoneal metastasis, so weekly paclitaxel (PTX) therapy was carried out. After 2 courses, malignant ascites completely disappeared and bilateral hydronephrosis improved. After 4 courses, no ascites or hydronephrosis were seen. Only neutropenia (grade 2) and alopecia (grade 1) were observed as adverse events during the therapy, but no major adverse events were noted. We also investigated the concentration of paclitaxel in ascites. Two hours after intravenous injection of PTX, the concentration of PTX in ascites rose over the reported cytotoxic dose of PTX, and this available concentration was maintained after 48 hours. Weekly paclitaxel therapy is suggested to be one of the safe and useful treatments for advanced gastric cancer with malignant ascites. PMID- 17108716 TI - [Thymoma--clinical aspects and its biological function]. AB - Thymoma is a neoplasm of thymic epithelial cells. The tumor grows relatively slowly, and the prognosis is very good after surgical resection. However, therapy for advanced thymoma has not been established, and the effect of modern chemotherapy is being evaluated. Thymoma is unique in that 25-30% of the patients have associated myasthenia gravis with autoantibodies to acetycholine receptor. The malignant epithelial cells maintain the function of cortical epithelial cells and harbor non-neoplastic immature T cells of CD 3(low) CD 4(+)CD 8(+) phenotype. This function has a causative relationship with the associated myasthenia gravis. Also, the presence of CD 3(low) CD 4(+)CD 8(+) T cells is diagnostic of thymoma and can be utilized for rapid diagnosis using needle biopsy specimen. PMID- 17108717 TI - [Patient-included multi-disciplinary team approach in the outpatient chemotherapy]. AB - To reassure safety and comfort in providing cancer chemotherapy on outpatient basis, patients and medical staff must form a team to manage the treatment together. Many patients tend to receive treatment rather passively in Japan until now, thus intervention is necessary for such patients to undergo the treatments with better understanding of the possible treatment side effects as well as the ways to deal with them. As for the risk management and/or symptom management, it is important to encourage patients to improve their ability for self care and to heighten their awareness that they can indeed take part in their own management. Also, when specialist intervention is required, consultation must be sought immediately so that the problem can be resolved promptly. It is important for the patient and the medical staff to share the same goal to provide safe and comfortable treatment, and for the member of the team fulfilling their respective specialist roles toward that goal. PMID- 17108718 TI - [Team approach for treatment of patients with cancer, how to cooperate with staffs of other medical institutions--a recent trend in Japan]. AB - For the team approach to patients with cancer both of consistency of medical services and patient's satisfaction are important. Japanese health care reform planning prescribes an establishment of a section which accepts patient's consultation and provides proper advice or information. Technology of the internet is also promising for team approaches with staffs of other medical institutions as well as the patient support section in the cancer centers. PMID- 17108719 TI - [Clinical pathways in the treatment of cancers]. AB - At the beginning, a clinical pathway was first introduced for the purpose of standardizing the treatment, providing high quality management and shortening hospital stays in the managed care health system in USA. This has been rapidly accepted also in Japan. The clinical pathway has also been gradually introduced in the treatment of malignant diseases, including chemo/radiotherapy and surgery. In Kurume University Hospital, clinical pathways are now applied for 50% of inpatients, and those for cancer patients have a large role in routine clinical practice. The progress of our pathways for cancer patients is here reported. PMID- 17108720 TI - [The role in cancer chemotherapy of "a pharmacist specialized in cancer"]. AB - A pharmacist system specialized in cancer has been established to authorize a clinical pharmacist to provide safe and sure support for cancer chemotherapy. A pharmacist specialized in cancer supports a protocol that has became complicated or involves management of anticancer agent side effects from a professional standpoint. The examination certifying a pharmacist specialized in cancer is very difficult, sophisticated (certification by pharmacists society) and requiring first-hand clinical experience of cancer chemotherapy to be eligible necessary for the examination. 41 pharmacists specialized in cancer were qualified in March of 2006. PMID- 17108721 TI - [Cancer care with informational technology]. AB - New innovations in clinical information technology have brought about changes in cancer care. The high-speed Internet provides an enormous real-time information exchange among physicians anywhere on earth. Computer-aided clinical decision support systems free up cancer experts in all stages of the cancer care continuum, including prevention, early detection, treatment and end-of-life care. Large registrations of cancer patients using the Internet enable us to construct large nationwide phenotypic and genotypic databases that allow clinicians to optimize individualized cancer treatment. Electronical medical records with an order-entry-system showing realtime clinical information provide safe, efficient and effective care. While new internet/intranet technologies reduce the inefficiencies of cancer care, the unintended consequences of the new informational technologies result in delivery of enormous unregulated clinical information, creating a digital divide between those using and not using the Internet, causing a decrease in communication. PMID- 17108722 TI - [Clinical significance of pyrimidine nucleoside phosphorylase (PyNPase) in breast cancer]. AB - To clarify the clinical significance of pyrimidine nucleoside phosphorylase (PyNPase) activity in breast cancer, we examined the possible correlation of PyNPase activity to clinicopathological features and prognosis in twenty-one patients with primary breast cancer from April 2000 to December 2001. Flow signals of tumors were analyzed by Power Doppler sonography (PDUS), and maximal velocity (V(max)) was calculated. PyNPase activity of resected specimens was assayed by ELISA method. PyNPase activities in resected cancerous tissue were 156.9+/-63.5 unit/mg (mean+/-SD), which were significantly higher than that in normal tissue (19.0+/-18.1 unit/mg, p<0.0001). PyNPase activity was positively correlated with tumor size (r=0.496, p=0.026) and V(max) (r=0.498, p=0.021). The disease free survival rate was significantly lower in the high PyNPase activity group than in the low PyNPase activity group. In overall survival rate, there was no significant difference between the high and low PyNPase activity groups. In the multivariate analysis, PyNPase activity was an independent predictor of postoperative recurrence (p=0.032). We suggest that PyNPase activity is associated with progression and proliferation of breast cancer, and that it may be useful for prediction of the prognosis. PMID- 17108723 TI - [Efficacy and toxicity of combination treatment with epirubicin (EPI) plus docetaxel (DOC) in advanced breast cancer]. AB - Thirty women (mean age 50.4 years, range 31-63) with primary advanced breast cancer were given EPI 40-60 mg/m(2) and DOC 50-60 mg/m(2) intravenously every three weeks. The efficacy was evaluated after 4 cycle treatments. There were 5 complete responses (CR) and 14 partial responses (PR), giving an overall response rate of 63.3%. There were 2 pathological CR (8%) which showed complete disappearance of cancer cells. The high dose group showed a better response than the low-dose group. The most common grade 3/4 adverse events were neutropenia (26.7%) and general fatigue (6.7%). The simultaneous combination treatment of EPI and DOC is effective for primary chemotherapy and can be performed safely even for outpatients. PMID- 17108724 TI - [A feasibility study of doxorubicin/cisplatin (AP) for postoperative chemotherapy in patients with advanced endometrial cancer]. AB - OBJECTIVE: We evaluated the feasibility of doxorubicin/cisplatin (AP) for postoperative chemotherapy in patients with advanced endometrial cancer. METHODS: Patients with newly diagnosed advanced endometrial cancer received AP (doxorubicin 60 mg/m(2), cisplatin 50 mg/m(2)) every 3 weeks. Treatment was continued until disease progression or completion of 6 courses. Toxicities were evaluated every cycle according to NCI-CTCAE Ver.3.0. RESULTS: Fifteen patients were enrolled from April 2004 through December 2005. All patients successfully completed therapy. There were two patients who needed dose reduction and nine patients with prolongation of treatment interval. Patients with over Grade 3/4 toxicity were observed to have leucopenia (47%), neutropenia (67%), anemia (26%), and vomiting (13%). No grade 3/4 cardiac and renal failure were observed. CONCLUSIONS: The doxorubicin/cisplatin (AP) regimen is tolerated and can be safely given without severe toxicity. PMID- 17108725 TI - Prognostic factors in patients with inoperable non-small cell lung cancer--an analysis of long-term survival patients. AB - A survival benefit of patients with inoperable non-small cell lung cancer has been reported since a development of new therapeutic agents in the 1990's. However, multivariate analyses of patients have not been evaluated. The aim of this study is to identify prognostic factors in the long-term survivors who had been treated with chemotherapy using these new agents and/or radiotherapy. A retrospective study and clinical analysis of 121 inoperable nonsmall cell lung cancer patients were conducted. Fifteen cases (male: 9, female: 6) with a survival of more than 2 years were revealed. Regarding clinical variables between the 15 cases and others, an early nodal (N) status, a high serum protein level, a good performance status (PS) and those having first-line chemotherapy or radiotherapy were all identified as significant prognostic factors for the long term survivors. Multivariate analyses also revealed that an early N status, a good PS, female gender and chemotherapy and/or radiotherapy were associated with the long-term survivors. These results suggest that patients with inoperable non small cell lung cancer should be considered for appropriate treatments including new chemotherapeutic agents. PMID- 17108726 TI - [Prediction of sensitivity to 5-fluorouracil (5-fu) by metabolic and target enzyme activities in colon cancer]. AB - BACKGROUND: 5-Fluorouracil (5-FU) and its derivatives are widely known as some of the most commonly prescribed anticancer drugs, especially for gastrointestinal cancer. Orotate phosphoribosyl transferase (OPRT), dihydropyrimidine dehydrogenase (DPD) and thymidylate synthase (TS) are initial key enzymes in the 5-FU metabolic pathway. The activities of these enzymes may have the potential to affect the chemosensitivity of 5-FU. PURPOSE: This study was designed to investigate the effect of OPRT, DPD and TS in sensitivity to 5-FU. METHOD: We measured OPRT, DPD and TS activities in 11 colonic cancer tissues. The Collagen Gel Droplet Embedded Culture Drug Sensitivity Test (CD-DST) was used in an in vitro chemosensitivity assay. In these samples, the correlation between sensitivity to 5-FU and enzyme activities was investigated. RESULTS: There were no correlations among OPRT, TS activities and sensitivity to 5-FU. In contrast, there was a significant inverse correlation (r=-0.738) between DPD activity and 5 FU sensitivity. With regression analysis, the coefficient of determination of the activity of the three enzymes versus the sensitivity to 5-FU was 0.61. CONCLUSION: Though measuring OPRT, DPD, TS activities is valuable for prediction of sensitivity to 5-FU, DPD is considered to be the most important predictive factor of 5-FU sensitivity. To improve its accuracy, the finding of a fourth factor such as P-glycoprotein and multidrug resistance-associated proteins (MRP), to be added to OPRT, DPD and TS, is desired. PMID- 17108727 TI - [A case of non-small cell lung cancer in which complete response was achieved with chemotherapy including cisplatin, vinorelbine, mitomycin C, followed by additional cisplatin, etoposide and concurrent radiotherapy]. AB - A 43-year-old man presented at our hospital with a complaint of cough and sputum. A plain chest X-ray and CT scan revealed a tumor shadow 8 cm in size in the right hilar and enlarged mediastinal lymph node. The tumor had invaded the superior vena cava. A tumor biopsy done under bronchoscopy revealed poorly-differentiated adenocarcinoma (cT4N2M0). He was given three courses of a combination therapy consisting of cisplatin (80 mg/m(2)), vinorelbine (25 mg/m(2)) and mitomycin C (8 mg/m(2)). Additionally, concurrent chemoradiotherapy (cisplatin 80 mg/m(2)+etoposide 100 mg/m(2), and 45 Gy) was performed. Right pneumonectomy was performed, because the primary tumor and the enlarged lymph node were markedly reduced in size, and a histological examination of the resected specimen revealed no detectable cancer cells. PMID- 17108728 TI - [An elderly patient with non-resectable lung cancer responding to TS-1]. AB - A 76-year-old man had complained of hoarseness, productive cough and left chest pain in June 2005. Chest radiograph and computed tomography showed a tumor 7.5x5 cm in size which contacted the aortic arch, some nodules in left upper lobe and right lower lobe of lung and left pleural effusion. The broncho-fiberscopic biopsy histologically proved the tumor to be moderately differentiated squamous cell carcinoma in the left upper lobe of lung. The patient was diagnosed as a non resectable lung cancer with clinical stage IV (c-T4N2M1). In July 2005, he was treated with oral administration of TS-1 (120 mg/day) and Jyu-zen daiho toh (7.5 g/day). After one week, he complained of grade 2 leukopenia and grade 1 skin rash. Then TS-1 was reduced to 100 mg/day. After this modulation of the treatment, he could take TS-1 and Jyu-zen daiho toh days 1-14, every 3 weeks. The patient then received 17 courses until June 2006. The left pleural effusion and left chest pain disappeared only 2 weeks after taking TS-1. In June of 2006, central necrosis of the left lung tumor was confirmed by positron emission computed tomography. No serious adverse effect was observed, and the patient maintained good quality of life. This case suggests that TS-1 with Jyu-zen daiho toh may be an effective treatment for elderly advanced lung cancer. PMID- 17108729 TI - [Complete resection of Pancoast tumor while receiving preoperative concurrent chemoradiotherapy (CCRT) as an induction therapy--report of a case]. AB - A 60-year-old man complaining of right shoulder pain and numbness of right arm was diagnosed with Pancoast tumor (invasive right apical lung cancer). Chest CT scan showed a tumor, 5 cm in diameter, in the right apex invading the right posterior chest wall. The patient received preoperative CCRT (RT: 40 Gy/20 Fr, cisplatin: CDDP and etoposide: ETP), resulting in tumor regression (PR). The patient underwent right upper lobectomy (ND 2a), partial resection of the 1st-3rd ribs and Th 1 nerve. Pathological examination demonstrated no live cancer cells and organization of necrotic tissue in the lung and intercostal region (Ef. 3). The patient received postoperative chemotherapy (CDDP+ETP) and was discharged. He did well without any tumor recurrence for 1 year postoperatively. CCRT seems effective and is one of the standard treatments for Pancoast tumor. PMID- 17108730 TI - [Results of hypotonic cisplatin treatment on seven patients with carcinomatous pleuritis]. AB - Carcinomatous pleuritis frequently undermines the quality of life in advanced malignant disease patients. The traditional treatment for pleural effusion is intrapleural administration of a chemical agent, but its efficacy was not at all satisfactory. Ichinose et al.developed hypotonic cisplatin treatment for patients with carcinomatous pleuritis in 1993, and its therapeutic efficacy was assessed by the Japan Clinical Oncology Group. This treatment was performed on seven patients in our hospital since 2002: five patients with non-small-cell lung cancer, one with transverse colon cancer who had myocardial infarction (grade 4) after treatment, and another with gastric cancer who had grade 4 hemosputa. These patients, aged 52 to 88, complained of dyspnea before treatment, but all of them were able to be discharged with control of pleural effusion including the above mentioned two patients. Although six patients had already died of disease, survival time after treatment of all patients ranged from 96 to 610 days, with an average of 346.7+/-228.2 days. The average time in five patients with non-small cell lung cancer was longer than that in two with gastrointestinal malignant disease (not significant statistically). Hypotonic cisplatin treatment seemed to enhance the quality of life in lung cancer patients with carcinomatous pleuritis. PMID- 17108731 TI - [Successful treatment of an elderly patient with advanced gastric cancer using low-doses of 5-fluorouracil, levofolinate calcium, and cis-platinum with chronomodulation]. AB - An 83-year-old man was admitted to our hospital for abdominal pain and severe vomiting. Gastrography and gastric endoscopy revealed that the pylorus was obstructed by a giant type 2 cancer in the lower gastric body. Furthermore, computed tomography revealed multiple metastases in the para-aortic lymph nodes. The man was unable to consume any food or liquid, and could not take medicine orally. Paclitaxel was not effective in treating the lymph node metastases or vomiting; therefore a chronomodulated schedule was used, which involved the administration of low doses of 5-fluorouracil, levofolinate calcium and cis platinum (FLP) each night. After four cycles of low-dose FLP therapy, the patient was able to consume food orally. The patient has partially responded to this regime over five months. In the current study, low-dose FLP therapy with chronomodulation was considered an effective treatment, and there were no severe adverse side effects. This case suggested that low-dose FLP therapy with chronomodulation is promising for the treatment of gastric cancer in elderly patients who can not take medicine orally, and further trials are warranted. PMID- 17108732 TI - [A case of long-term survival of 3-years 4 months after combination chemotherapy of MTX, 5-FU and low-dose CDDP (MFP) for type 4 gastric cancer with pleuritis, peritoneal dissemination and Krukenberg tumor]. AB - A 41-year-old woman presented to the Department of Obstetrics and Gynecology of our hospital because of abdominal distension and irregular genital bleeding. Computed tomography and ultrasonography of the abdomen revealed bilateral ovarian tumors, massive ascites, and bilateral pleural effusion. Type IV advanced gastric cancer was diagnosed on upper gastrointestinal endoscopy. The patient was admitted to our department. She received 3 courses of combination chemotherapy with methotrexate, 5-fluorouracil, and low-dose cisplatin. Pleural effusion and ascites disappeared. Surgery (total gastrectomy, resection of the tail of the pancreas, lymph-node dissection, total hysterectomy, and adnexectomy) was performed, and the patient was discharged. Chemotherapy was repeated after surgery. Lymph-node metastasis recurred 1 year 8 months after the start of chemotherapy. Treatment was switched to irinotecan plus cisplatin, and the lymph nodes shrank. After 9 months, 3 courses of TS-1 were administered. Two years 10 months after starting chemotherapy, abdominal and low back pain developed. Bone scintigraphy revealed bone metastasis. Lymph node swelling was present. The patient responded to radiotherapy with chemotherapy (cisplatin plus 5 fluorouracil). Subsequently, abdominal computed tomography showed lymph-node swelling, multiple metastases to the liver, ascites, and a right pleural effusion. She was readmitted to the hospital and received intraperitoneal chemotherapy with cisplatin. Her condition deteriorated, and she died. The patient survived for about 3 years 4 months after the start of treatment. Chemotherapy with methotrexate, 5-fluorouracil, and low-dose cisplatin may thus be an effective therapeutic option in patients who have advanced gastric cancer with peritoneal dissemination. PMID- 17108733 TI - [Three cases of successfully resectable advanced gastric cancer responding to TS 1 plus CDDP]. AB - We have experienced three gastric cancer cases successfully treated by the combination therapy of CDDP and TS-1. Case 1: 65-year-old man, a type 4 gastric cancer with huge abdominal para-aortic lymphnodes. Case 2: 74-year-old woman, a type 3 gastric cancer was found with obstructive jaundice. Swelling lymph nodes were seen in the hepatoduodenal ligament. Case 3: 58-year-old woman, a type 4 gastric cancer was found. The tumor invading the pancreas and swelling lymphnodes around the stomach were confirmed according to abdominal CT. Although combined chemotherapy of TS-1 plus CDDP is effective for advanced gastric cancer,adjuvant chemotherapy must be investigated in noneffective cases. PMID- 17108734 TI - [A case of perforated gastric cancer in which complete response was confirmed with resection following TS-1/CDDP combined chemotherapy after omentopexy]. AB - The present patient was a 50-year-old male with sudden upper abdominal pain. The patient exhibited pallor, and physical examination revealed a rigid abdomen. Abdominal x-ray revealed free air, and emergency laparotomy was performed to confirm upper gastrointestinal tract perforation. A perforated lesion of approximately 1 cm in diameter was found on the anterior wall at the gastric angle. The area surrounding the lesion was tumor-like, and the posterior wall was fused invasively with the pancreas. Malignancy was suspected; however, considering the patient's general status, greater omentum grafts were opted for. The patient was diagnosed with type III gastric cancer by gastroendoscopy post operatively, and TS-1/CDDP therapy was started on the 28th day after surgery. After three courses of treatment, the tumor was found to have smoothened, wall consolidation was improved, and a second surgery was performed. During laparotomy, a nodule in the round ligament of liver was found and removed; however, there were no other medical findings that raised suspicion of peritoneal dissemination or liver metastasis. It was concluded that radical surgery was possible, and distal gastrectomy (D 2) was performed. Pathological examination revealed that signet ring cell carcinoma was present on only part of the mucous membrane. The lower and muscle layers of the serous membrane and the nodule in the round ligament of liver were replaced by fibrous tissue, indicating the disappearance of cancer cells. Two years and three months after surgery, the patient exhibited no signs of recurrence. PMID- 17108735 TI - [A case of gemcitabine-based chemo-radiation therapy for locally advanced unresectable pancreatic cancer]. AB - A 49-year-old man was admitted to our hospital with vomiting. Abdominal CT revealed an avascular tumor at the uncinate process of the pancreas measuring 36x30 mm. Preoperative serum CA 19-9 was 361 U/ml. During laparotomy,the tumor was deemed unresectable (T4NXM0, Stage IVa),and duodenojejunostomy was performed. External-beam radiotherapy (EBRT) (50.4 Gy/28Fr) with concurrent twice-weekly gemcitabine (GEM) (40 mg/m(2)/day) was delivered. In the outpatient setting, and 1,000 mg/m(2) of GEM was administered intravenously on days 1, 8, and 15. Cycles were repeated every 28 days. The patient received 13 cycles of GEM chemotherapy until the appearance of a grade 2 facial rash. A decrease in tumor size was observed, and the serum CA 19-9 level dropped to 16 U/ml. He remained well without any symptoms and pursued normal activity for 33 months. He died of peritoneal dissemination 43 months after diagnosis. Gemcitabine-based chemo radiation seems to be a safe and effective regimen for unresectable pancreatic cancer. PMID- 17108736 TI - [A case of intrahepatic cholangiocarcinoma effectively treated by hepatic arterial infusion chemotherapy]. AB - The patient was a 50-year-old woman who suffered from gastric discomfort. She was first diagnosed as intrahepatic cholangiocarcinoma with hepatic, paraaortic lymphnodal and bone metastasis. Initial systemic chemotherapy using gemcitabine (GEM) and 5-FU failed to control the disease activity. Then she was given GEM and cisplatin (CDDP) combination chemotherapy. The response was assessed as stable disease (SD), but grade 4 leukopenia was seen. Then systemic therapy using GEM, and hepatic arterial infusion therapy with CDDP, l-leucovorin and 5-FU were continued biweekly. Partial response (PR) was achieved six months later, and her disease status was maintained as SD. This hepatic arterial infusion chemotherapy would be safe and feasible as therapy for inoperable intrahepatic cholangiocarcinoma. PMID- 17108737 TI - [A case of duodenal carcinoma with massive peritoneal carcinomatosis complicating a pulmonary embolism]. AB - The incidence of venous thromboembolism in cancer patients is reportedly around 15%, and a pulmonary embolism is an important cause of morbidity and mortality. We present a case of duodenal carcinoma with peritoneal carcinomatosis complicated by a pulmonary embolism. Heparin, followed by warfarin, was useful to treat the embolism. The combination chemotherapy with TS-1 and docetaxel was effective for the peritoneal carcinomatosis. The QOL of this patient was maintained for about 5 months. PMID- 17108738 TI - [Cancer board--new medical system in cancer clinic]. AB - To proceed with more patient-oriented cancer medicine, the development of a new medical system is necessary in Japan. In Ariake Hospital of the Japanese Foundation for Cancer Research, a new medical system has been developed, which is composed of a common outpatient clinic and a conference system. The core conference system is called "Cancer Board". The practice of patient-oriented medicine in gastroenterological cancer has been introduced as a sample of the Cancer Board system in our hospital. PMID- 17108739 TI - [A case of advanced rectal carcinoma with multiple lung metastases responding to irinotecan combined with 5-fluorouracil and l-leucovorin (IFL) as neoadjuvant chemotherapy (NAC)]. AB - A 53-year-old man, admitted for inguinal hernia, complained of body weight loss in a preoperative condition check. We examined the digestive tract and diagnosed stage IV advanced rectal carcinoma with multiple lung metastases. It caused ileus, so emergency colostomy was performed. After that his general condition recovered, and two cycles of neoadjuvant chemotherapy (NAC) by irinotecan combined with 5-fluorouracil and l-leucovorin (IFL) therapy were performed on an outpatient basis. Lung metastatic nodules disappeared. We established a diagnosis of down staging for stage IIIa, and performed a lower anterior resection with D 2 lymph node dissection to allow a curability-A resection. The pathological effect of NAC was Grade 2. Post-operatively, two cycles of IFL therapy were then performed. There has been no sign of recurrence, and no adverse effects by chemotherapy have been seen during this treatment. Thus, NAC by IFL therapy can be one of the useful treatment approaches for patients with advanced rectal cancer. PMID- 17108740 TI - [Partial response of liver metastasis from colon cancer following intermittent oral administration of TS-1 after intra-hepatic arterial single injection of CDDP, doxorubicin and 5-FU]. AB - An intrahepatic arterial injection of CDDP, doxorubicin and 5-FU, followed by 17 courses of oral TS-1 administration (80 mg/day x 14 days, q=28 days), induced a partial response for 9 months after 18 months of stable disease in a 76-year-old male with asynchronous liver metastasis due to ascending colon cancer. TS-1 showed an excellent anticancer effect against colorectal metastatic liver cancers for a long time without loss of QOL or safety. PMID- 17108741 TI - [A case of follicular lymphoma complicated with lethal pemphigus]. AB - Paraneoplastic pemphigus (PNP) is a rare autoimmune bullous disease associated with neoplasm, which is clinically characterized by mucocutaneous lesions resembling pemphigus vulgaris or erythema multiforme. A case presented with PNP refractory to chemotherapy including rituximab, predonisolone and cyclophosphamide (RCHOP regimen). A 36-year-old man, who had been diagnosed as extended follicular lymphoma, presented with a polymorphous skin eruption of the trunk, sclera conjunctivitis, and severe mucosal erosions of the lips and oral cavity. He was diagnosed as pemphigus pathologically by a biopsy of the oral mucosa. However, 3 courses of rituximab and CHOP therapy, which exert a partial response with lymphoma lesions, did not prove effective for oral stomatitis due to pemphigus. He received corticosteroid therapy (prednisolone 40 mg/day) and went into a state of temporally remission regarding pemphigus. However, the mucosal lesions were again exacerbated despite control of the lymphoma status after chemotherapy. Oral stomatitis extended to the upper respiratory system through the larynx and resulted in bronchiolitis obliterance clinically presented likely as severe chronic obstructive pulmonary disease (COPD). Because it is known that PNP refractory to long-term steroid and cytoreductive therapy has a progressive character and poor prognosis, supportive care would be warranted for these patients. PMID- 17108742 TI - [What do cancer chemotherapy outpatients want?--results of a questionnaire survey]. AB - We conducted a questionnaire survey to evaluate patients at the cancer chemotherapy center of the university of occupational and environmental health hospital from January to March 2006. Thirty-three (61.1%) and 5 (9.2%) of 54 patients wished to receive chemotherapy on an outpatient basis and as inpatients, respectively. Among the reasons for wishing to be treated as outpatients were the following: Keeping in touch with family, not wanting to be hospitalized, a desire to work, and the wish to continue hobbies, at 45.5 (15/33), 36.4 (12/33), 24.2 (8/33), and 24.2 (8/33), respectively. Twenty-four (44.4%) patients voiced concerns about chemotherapy at an outpatient clinic in terms of associated adverse events, feeling bad about causing trouble to their family, and the desire to have consultations at night or on holidays. Thirty-eight (70.4%) patients wished to have chemotherapy within 3 hours, and 42 (77.7%) of them requested improvements in the examinations, time, duration of chemotherapy, and the effect of chemotherapy. These findings indicate that a major part of patients wish to receive cancer chemotherapy at an outpatient clinic within 3 hours. However, they also have serious reservations about any associate adverse events. We therefore need to improve the overall environment for cancer chemotherapy patients in order to allow them to feel safe when undergoing such treatments. PMID- 17108743 TI - [What happened during these 12 years since CPT-11 was launched in Japan ?]. AB - CPT-11 (irinotecan hydrochloride, trade name: Campto or Topotecin) was launched in 1994. L-OHP (oxaliplatin, trade name: Elplat) was approved based on the fast track evaluation system and launched in 2005. Originally, both of these drugs were synthesized in Japan. Just after the launch of CPT-11, the severity of its toxicity was reported more frequently than its efficacy, therefore it spent much time to spread the use of this drug. As for L-OHP, the approved regimen is FOLFOX in spite the regimen was not actually studied in its registration studies in Japan. However, L-OHP is widely used after its launch. Thus, we find a progress in terms of regulatory system to introduce the widely accepted standard chemotherapy to the Japanese practice sites rapidly. We also find a further understanding for cancer chemotherapy in Japanese society. Recently, mass media reported cancer patients who were eager to receive the standard chemotherapy and requesting the regulatory authorities its quick approval. We have never seen such a scene 10 years ago. These patients' activities could be a key factor to change the infrastructure of cancer therapy. PMID- 17108745 TI - Preface. PMID- 17108744 TI - Contributing authors. PMID- 17108749 TI - Community hospital successfully implements eRecord and CPOE. AB - Despite media attention on converting the nation's paper-based medical record systems to electronic systems, few hospitals, and even fewer community hospitals, have done so. University of Pittsburgh Medical Center St. Margaret has converted to a comprehensive electronic health record system, known as eRecord, in a short time. The authors describe key factors that were critical to the success of the conversion, along with positive results on quality of care. PMID- 17108750 TI - Using synchronous software in Web-based nursing courses. AB - To promote learning and enhance immediacy and satisfaction, a college of nursing at a large research I southern university undertook a pilot project to incorporate synchronous classroom software into an ongoing online program. Two synchronous class sessions using voice over Internet protocol were offered in the Community/Public Health Nursing course through Elluminate Live! Upon conclusion of the lecture, students were divided into breakout groups to work on group projects. Surveys were administered to the students and faculty before and after the class sessions. Evaluation of the pedagogical strategies used in the synchronous sessions was conducted by instructional technology faculty. Students in the pilot group reported higher levels of satisfaction with the Web-based course with synchronous sessions. In addition, students reported that group time at the end of the session was helpful for completing group projects. A majority responded that synchronous session activities and assignments facilitated their understanding of course content. This article presents a description of the synchronous classroom pilot project along with recommendations for implementation and pedagogical approaches. PMID- 17108752 TI - Assistive computing devices: a pilot study to explore nurses' preferences and needs. AB - Healthcare errors among nurses are common because of the fast-paced work environment, in which extensive data must be analyzed and quick decision making is required. Assistive computing devices can help reduce nursing errors by providing timely access to client information and by assisting nurses with client monitoring, decision making, and bedside documentation. The purpose of this pilot study was to determine what assistive computing device features, functions, and input/output modalities nurses would find most useful as support for their nursing duties. Twenty nurses completed a questionnaire that examined their needs and preferences. Data analysis revealed a strong desire for capabilities related to facilitating information access and administering safe medication. The results of this study will be used to develop design criteria for an assistive computing device that will aim to improve the performance of nurses through appropriate information and data support. PMID- 17108753 TI - Utility of a Web-based intervention for individuals with type 2 diabetes: the impact on physical activity levels and glycemic control. AB - Despite the numerous benefits of physical activity for patients with diabetes, most healthcare providers in busy clinical settings rarely find time to counsel their patients about it. A Web-based program for healthcare providers can be used as an effective counseling tool, when strategies are outlined for specific stages of readiness for physical activity. Seventy-three adults with type 2 diabetes were randomly assigned to Web-based intervention, printed-material intervention, or usual care. After 12 weeks, the effects of the interventions on physical activity, fasting blood sugar, and glycosylated hemoglobin were evaluated. Both Web-based and printed material intervention, compared with usual care, were effective in increasing physical activity (P < .001) and decreasing fasting blood sugar (P<.01) and glycosylated hemoglobin (P < .01). Post hoc analysis for change scores indicated significant differences between Web-based intervention and usual care and between printed material intervention and usual care, but not between web-based and printed material intervention. The findings of this study support the value of Web-based and printed material interventions in healthcare counseling. With increasing Web access, the effectiveness of Web-based programs offered directly to patients needs to be tested. PMID- 17108754 TI - Mapping of nursing records into the NIC and the ICNP in a Korean oriental medicine hospital. AB - This study uses mapping methodology to examine the applicability of the Nursing Interventions Classification and the International Classification of Nursing Practice to nursing practice in a Korean Oriental-medicine hospital. Data were collected from the nursing records of 56 stroke patients in one unit, and intervention statements were mapped into NIC and ICNP. Of 147 unique nursing intervention statements extracted, 136 (92.52%) could be mapped into NIC and 99 (67.35%) statements could be completely mapped into ICNP. Using mapping methodology, this study validates that both NIC and ICNP would be useful for documenting nursing care in a Korean hospital, but it also identifies additional concepts that need to be represented in both of these standardized nursing languages. It is recommended that nurses be more careful in documenting their interventions and also that SNLs be developed further to more completely represent nursing practice. PMID- 17108755 TI - Surveying acute care providers in the US to explore the impact of health information technology on the role of nurses and interdisciplinary communication in acute care settings. PMID- 17108758 TI - Fifteen years of experience in predictive testing for Huntington disease at a single testing center in Victoria, Australia. AB - PURPOSE: This retrospective study describes 15 years of experience in predictive testing for Huntington disease at a single center in Victoria, Australia. METHOD: Data collected on 756 participants included age, gender, family history, prior risk and the age at which this risk became known, exposure to Huntington disease, number of children, and proximity to the testing center. RESULTS: Some 57.8% of participants were female, and 88.8% had a 50% risk of developing Huntington disease. The mean age at entry was 40.4 years and was gender-independent. Of all completed tests (n = 648), 37.5% gave high-risk results, and 3.2% were in the zone of reduced penetrance. The 14.3% who withdrew from testing tended to be younger and childless, lacked exposure to severe Huntington disease, and more often at 25% or less risk. Some 32.4% of candidates presented for testing within 1 year of becoming aware of their risk, and most of these individuals had little or no exposure to severe Huntington disease. Those whose exposure was considerable waited on average for more than 13 years. Among the most inexperienced candidates were a group of "adoptees" (raised away from their biological family). Maternal transmission was the source of risk for 19 of 20 adoptees. CONCLUSION: This study illustrates the significance of exposure to Huntington disease and its impact on the timing of testing. PMID- 17108759 TI - Impact of hemochromatosis screening in patients with indeterminate results: the hemochromatosis and iron overload screening study. AB - PURPOSE: Assess the quality of life impact of receiving indeterminate test results for hemochromatosis, a disorder involving HFE genetic mutations and/or elevated serum transferrin saturation and ferritin. METHODS: The study sample was from the Hemochromatosis and Iron Overload Screening Study, a large observational study of hemochromatosis among primary care patients in the US and Canada using HFE genotype and serum transferrin saturation and ferritin screening. Study subjects included 2,304 patients found with hemochromatosis risk of uncertain clinical significance. Assessed was SF-36 general health and emotional well-being before screening and six weeks after participants received their test results. Health worries were assessed after screening. RESULTS: Of the study subjects, 1,268 participants (51.5%) completed both assessments. Compared to normal controls, those with HFE mutations or elevated serum transferrin saturation and ferritin levels of uncertain significance were more likely to report diminished general health and mental well-being, and more health worries. These effects were associated with participants' belief of having tested positive for hemochromatosis or iron overload. CONCLUSION: Notification of indeterminate results from screening may be associated with mild negative effects on well being, and might be a potential participant risk in screening programs for disorders with uncertain genotype-phenotype. PMID- 17108760 TI - Risk communication in completed series of breast cancer genetic counseling visits. AB - PURPOSE: There is no consensus on how best to communicate risk in breast cancer genetic counseling. We studied risk communication in completed series of counseling visits and assessed associations with counselees' postcounseling risk perception and satisfaction. METHODS: Pre- and postcounseling questionnaires and videorecordings of all visits were available for 51 affected and unaffected women from families with no known BRCA1/2 mutation, who fulfilled criteria for DNA testing. We developed a checklist for assessing risk communication and counselors' behaviors. RESULTS: General risks were mainly communicated in initial visits, while counselee-specific risks were discussed mainly in concluding visits. The risks discussed most often were conveyed only numerically or qualitatively, and most were only stated positively or negatively. Counselors regularly helped counselees to understand the information, but seldom built on counselees' pre-existing perspective. Counselees' breast cancer risk perception after counseling was unrelated to whether this risk had been explicitly stated. The number of general risks discussed was negatively associated with counselees' satisfaction about counseling. CONCLUSION: Findings suggest that counselors' authority prevails over mutuality with individual counselees, in their communication about risks. PMID- 17108761 TI - Impact of familial adenomatous polyposis on young adults: attitudes toward genetic testing, support, and information needs. AB - PURPOSE: The study assessed views concerning genetic testing and information and support needs among young adults aged 18 to 35 years with a diagnosis of or at risk of developing familial adenomatous polyposis. METHODS: A total of 88 participants were recruited through Hereditary Bowel Cancer Registries and assessed using self-administered questionnaires. RESULTS: The average age of participants was 28 years, and the average age at the time of their last genetic consultation was 23 years. Although 75% would consider prenatal genetic testing, only 21% would consider termination of an affected pregnancy. Sixty-one percent selected "at birth" or "early childhood" as the preferred age for genetic testing of offspring. Participants' highest areas of unmet support needs were with regard to anxiety about their children having familial adenomatous polyposis (39%) and fear of developing cancer (28%). CONCLUSION: The parental desire to test children before it is clinically indicated may be a source of distress and create conflict with genetic services. These findings demonstrate that familial adenomatous polyposis may significantly impact young adults, with many having unmet support needs. The length of time since the last genetic consultation and the young age at which these consultations took place suggest that clinics should consider a means of regular follow-up to address these unmet needs. PMID- 17108762 TI - A complex additive model of inheritance for Hirschsprung disease is supported by both RET mutations and predisposing RET haplotypes. AB - PURPOSE: The RET proto-oncogene is considered to be the major susceptibility gene involved in Hirschsprung disease. Traditional RET germline mutations account for a small subset of Hirschsprung disease patients, but several studies have shown that there is a specific haplotype of RET associated with the sporadic forms of Hirschsprung disease. We have investigated for RET germline mutations and analyzed the RET haplotypic distribution in carriers versus noncarriers of RET germline mutations. METHODS: We have screened the coding region of RET in 106 Spanish Hirschsprung disease patients using dHPLC technology. Statistical comparisons of the distribution of RET haplotypes between sporadic patients with and without a RET germline mutation were performed. RESULTS: Nine novel germline mutations and one previously described were identified. A significant over transmission of the "Hirschsprung disease haplotype" was detected when comparing transmitted versus nontransmitted alleles in the group of Hirschsprung disease triads without mutation. However, no distortion of the transmission of alleles was found in the group of mutated families. CONCLUSIONS: These results would be concordant with a complex additive model of inheritance. The whole findings seem to suggest that low-penetrance mutations would be necessary but not sufficient and the additional presence of the "Hirschsprung disease haplotype" could contribute to the manifestation of the disease. PMID- 17108763 TI - Functional copper transport explains neurologic sparing in occipital horn syndrome. AB - OBJECTIVE: A range of neurologic morbidity characterizes childhood-onset copper transport defects, including severe Menkes disease and milder occipital horn syndrome. Both phenotypes are caused by mutations in ATP7A, which encodes a copper-transporting adenosine triphosphatase, although defects causing occipital horn syndrome are rarely reported and nearly always involve exon-skipping (six of eight prior reports). Our objective was to characterize a novel occipital horn syndrome mutation (N1304S) not associated with aberrant splicing and to determine whether functional copper transport was associated with this allele. METHODS: We studied two brothers with typical occipital horn syndrome and used yeast complementation and timed growth assays, exploiting a Saccharomyces cerevisiae mutant strain, to assess in vitro N1304S copper transport. RESULTS: We documented that N1304S has approximately 33% residual copper transport, a result not inconsistent with a similar patient we reported with an exon-skipping mutation whose cells showed correctly spliced mRNA transcripts 36% of normal. CONCLUSION: These patients' mild neurologic phenotypes, together with our yeast complementation and growth experiments, imply that N1304S does not completely block copper transport to the developing brain early in life. The findings suggest that neurologic sparing in untreated occipital horn syndrome is associated with approximately 30% residual functional activity of ATP7A. PMID- 17108764 TI - Prenatal diagnosis of chromosomal abnormalities using array-based comparative genomic hybridization. AB - PURPOSE: This study was designed to evaluate the feasibility of using a targeted array-CGH strategy for prenatal diagnosis of genomic imbalances in a clinical setting of current pregnancies. METHODS: Women undergoing prenatal diagnosis were counseled and offered array-CGH (BCM V4.0) in addition to routine chromosome analysis. Array-CGH was performed with DNA directly from amniotic fluid cells with whole genome amplification, on chorionic villus samples with amplification as necessary, and on cultured cells without amplification. RESULTS: Ninety-eight pregnancies (56 amniotic fluid and 42 CVS specimens) were studied with complete concordance between karyotype and array results, including 5 positive cases with chromosomal abnormalities. There was complete concordance of array results for direct and cultured cell analysis in 57 cases tested by both methods. In 12 cases, the array detected copy number variation requiring testing of parental samples for optimal interpretation. Array-CGH results were available in an average of 6 and 16 days for direct and cultured cells, respectively. Patient acceptance of array-CGH testing was 74%. CONCLUSION: This study demonstrates the feasibility of using array-CGH for prenatal diagnosis, including reliance on direct analysis without culturing cells. Use of array-CGH should increase the detection of abnormalities relative to the risk, and is an option for an enhanced level of screening for chromosomal abnormalities in high risk pregnancies. PMID- 17108765 TI - Reliable detection of trisomy 21 using MALDI-TOF mass spectrometry. AB - PURPOSE: Current diagnostic methods for chromosomal abnormalities rely mainly on karyotyping and occasionally fluorescent in situ hybridization or quantitative polymerase chain reaction. We describe an alternative molecular method for the detection of trisomy 21 involving mass spectrometric analysis of single nucleotide polymorphisms. METHODS: In collaboration with Sequenom, Inc., 350 blinded amniotic fluid, amniocyte culture, chorionic villus, or amniotic fluid supernatant samples were analyzed for trisomy 21 using SNP analysis and matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Peak ratios were calculated for heterozygous genotypes and compared to control values generated from known euploid samples. An analytical algorithm using standard deviations from control values was used to determine the probability of a sample being affected or unaffected. RESULTS: Seventy-three trisomy 21 samples from among the 350 blinded samples were correctly identified. There were no false-positive or false-negative results among the complete trisomy 21 samples. One sample exhibiting mosaicism for trisomy 21 was identified as being unaffected. CONCLUSIONS: MALDI-TOF mass spectrometry is a robust and reproducible method for the detection of trisomy 21. Its amenability to high throughput analysis and high degree of multiplexing make it a potential future diagnostic tool for the detection of other aneuploidies as well. PMID- 17108768 TI - Searching the literature: understanding and using structured electronic databases. PMID- 17108769 TI - Home-prepared saline: a safe, cost-effective alternative for wound cleansing in home care. AB - OBJECTIVE: The purpose of this study was to determine (1) the rate of contamination for a normal saline solution prepared with distilled water and salt when stored at room temperature and (2) when stored under refrigerated conditions; and (3) to compare the rate of contamination for solutions prepared and stored under controlled conditions with those prepared and stored in patients' homes. DESIGN: This quasi-experimental study examined the bacterial content of saline made with distilled water and stored at room temperature and that of saline stored in a refrigerator. SETTING: A large urban home health agency was the setting for the study. METHODS: Eight teaspoons of table salt were added to 1-gallon jugs of distilled water purchased from a grocery store to produce a normal saline solution. Two gallons of the prepared solution were stored at room temperature (80 degrees F) and 2 were stored in a refrigerator (48 degrees F). Cultures were taken from each of the 4 bottles of prepared solution immediately following preparation and at weekly intervals until bacterial growth occurred, or 4 weeks after preparation. RESULTS: All tested solutions were negative for bacterial growth at baseline. Cultures of the solution stored at room temperature were positive for bacterial growth 2 weeks after preparation, whereas those stored under refrigerated conditions remained free of bacterial contamination 4 weeks after preparation. Two "patient-prepared" solutions were tested at 3 weeks, and both were negative for bacteria at that time. Three patient-prepared solutions were tested at 4 weeks, of which 2 showed no bacterial growth in 48 hours, whereas 1 revealed 600 cfu/mL of Pseudomonas species. CONCLUSIONS: The results of this small study suggest that saline solutions produced by adding table salt to distilled water will remain bacteria-free for 1 month if refrigerated. Additional studies are needed to confirm these findings. PMID- 17108770 TI - Nursing staff beliefs and expectations about continence care in nursing homes. AB - OBJECTIVE: The purpose of this qualitative study was to consider the current beliefs of nursing assistants and directors of nursing about management of urinary incontinence (UI) among the residents in nursing homes. DESIGN: This was a qualitative study using purposive sampling and a focus group methodology. SETTING AND SUBJECTS: Three focus groups including 38 participants were held in 2 different regions. Two of the focus groups comprised nursing assistants and 1 comprised directors of nursing. METHOD: The focus groups were facilitated by 2 different advanced practice nurses (BR and LJK), and 2 similar interview guides were used: 1 for the nursing assistants and 1 for the nurses. The interviews were tape recorded and transcribed verbatim; data were analyzed via content analysis. RESULTS: Ten themes were identified from the data; 3 focused on resident factors that influence UI, 4 related to staff and family contributors to UI, and 3 focused on recommendations to improve UI management in the nursing home setting. CONCLUSIONS: The findings from this study can be used to guide interventions to decrease or eliminate barriers to continence care and thereby facilitate the implementation of clinical practice guidelines and evidence-based interventions to improve urinary continence among nursing home residents. PMID- 17108771 TI - Urinary incontinence and diabetes: a review. AB - Both urinary incontinence and diabetes are chronic and costly conditions affecting millions of Americans. Unfortunately, these chronic conditions often coexist in the same person, compounding the economic and social costs and diminishing quality of life. The connection between urinary incontinence and diabetes has been mentioned in subsets of articles under prevalence or risk factors. The relationship is strong enough that researchers and clinicians should be aware of this link when formulating protocols or providing clinical care. This article summarizes knowledge of diabetes mellitus and urinary incontinence and explores the nature of their relationship. PMID- 17108772 TI - A clinical nursing leadership model for enhancing continence care for older adults in a subacute inpatient care setting. AB - This article describes the application and evaluation of a clinical nursing leadership model to enhance continence care in an inpatient rehabilitation setting for older adults. Multiple strategies were employed to optimize the uptake and sustainability of 3 practice initiatives: (1) establishment of an enhanced role for ward-based nurses to provide clinical support to patients and other staff for the management of incontinence, (2) a new method for nursing assessment and management of bowel elimination, and (3) a framework for improved discharge care of patients with incontinence. Evaluation data indicated a high level of acceptance of the role of the ward-based continence resource nurses, improved assessment and management of bowel elimination, and enhanced discharge care for patients with incontinence. These initiatives were sustainable during a 2-year period after their introduction, despite the gradual withdrawal of the clinical leader. PMID- 17108773 TI - Pharmacotherapeutics in older adults. AB - The process of aging influences both pharmacodynamics and pharmacokinetics. In addition to this, the issue of the increased incidence of chronic diseases as the age of people and the effects of medications in older adults becomes very complex. This article will review the influence of the aging process on the absorption, distribution, metabolism, and excretion of drugs. Specific concerns of older adults, including drug groups and side effects of concern, drug-induced geriatric syndromes, and medication adherence, are also discussed. PMID- 17108775 TI - Use of a clear absorbent acrylic dressing for debridement. AB - Outpatient wound care centers are often challenged with patients whose wounds have multiple etiologies. These patients require a plan of care that debrides devitalized tissue, promotes wound healing, and addresses underlying factors. Because of the chronic nature of these wounds and the time required for healing, clinicians practicing in ambulatory settings also must coordinate care and appreciate the unique challenges when wound management extends into the home healthcare setting. PMID- 17108778 TI - Serum gastrin in Zollinger-Ellison syndrome: I. Prospective study of fasting serum gastrin in 309 patients from the National Institutes of Health and comparison with 2229 cases from the literature. AB - The assessment of fasting serum gastrin (FSG) is essential for the diagnosis and management of patients with the Zollinger-Ellison syndrome (ZES). Although many studies have analyzed FSG levels in patients with gastrinoma, limited information has resulted from these studies because of their small size, different methodologies, and lack of correlations of FSG levels with clinical, laboratory, or tumor features in ZES patients. To address this issue, we report the results of a prospective National Institutes of Health (NIH) study of 309 patients with ZES and compare our results with those of 2229 ZES patients in 513 small series and case reports in the literature. In the NIH and literature ZES patients, normal FSG values were uncommon (0.3%-3%), as were very high FSG levels >100-fold normal (4.9%-9%). Two-thirds of gastrinoma patients had FSG values <10-fold normal that overlap with gastrin levels seen in more common conditions, like Helicobacter pylori infection or antral G-cell hyperplasia/hyperfunction. In these patients, FSG levels are not diagnostic of ZES, and gastrin provocative tests are needed to establish the diagnosis. Most clinical variables (multiple endocrine neoplasia type 1 status, presence or absence of the most common symptoms, prior medical treatment) are not correlated with FSG levels, while a good correlation of FSG values was found with other clinical features (prior gastric surgery, diarrhea, duration from onset to diagnosis). Increasing basal acid output, but not maximal acid output correlated closely with increasing FSG. Numerous tumoral features correlated with the magnitude of FSG in our study, including tumor location (pancreatic > duodenal), primary size (larger > smaller) and extent (liver metastases > local disease). In conclusion, this detailed analysis of FSG in a large number of patients with ZES allowed us to identify important clinical guidelines that should contribute to improved diagnosis and management of patients with ZES. PMID- 17108779 TI - Serum gastrin in Zollinger-Ellison syndrome: II. Prospective study of gastrin provocative testing in 293 patients from the National Institutes of Health and comparison with 537 cases from the literature. evaluation of diagnostic criteria, proposal of new criteria, and correlations with clinical and tumoral features. AB - In two-thirds of patients with Zollinger-Ellison syndrome (ZES), fasting serum gastrin (FSG) levels overlap with values seen in other conditions. In these patients, gastrin provocative tests are needed to establish the diagnosis of ZES. Whereas numerous gastrin provocative tests have been proposed, only the secretin, calcium, and meal tests are widely used today. Many studies have analyzed gastrin provocative test results in ZES, but they are limited by small patient numbers and methodologic differences. To address this issue, we report the results of a prospective National Institutes of Health (NIH) study of gastrin provocative tests in 293 patients with ZES and compare these data with those from 537 ZES and 462 non-ZES patients from the literature. In 97%-99% of gastrinoma patients, an increase in serum gastrin post secretin (Delta secretin) or post calcium (Delta calcium) occurred. In NIH ZES patients with <10-fold increase in FSG, the sensitivity/specificity of the widely used criteria were as follows: Delta secretin > or =200 pg/mL (83%/100%), Delta secretin >50% (86%/93%), Delta calcium > or =395 pg/mL (54%/100%), and Delta calcium >50% (78%/83%). A systematic analysis of the sensitivity and specificity of other possible criteria for a positive secretin or calcium test allowed us to identify a new criterion for secretin testing (Delta > or =120 pg/mL) with the highest sensitivity/specificity (94%/100%) and to confirm the commonly used criterion for calcium tests (Delta > or =395 pg/mL) (62%/100%). This analysis further showed that the secretin test was more sensitive than the calcium test (94% vs. 62%). Our results suggest that secretin stimulation should be used as the first-line provocative test because of its greater sensitivity and simplicity and lack of side effects. In ZES patients with a negative secretin test, 38%-50% have a positive calcium test. Therefore the calcium test should be considered in patients with a strong clinical suspicion of ZES but a negative secretin test. Furthermore, we found that some clinical (diarrhea, duration of medical treatment), laboratory (basal acid output), and tumoral (size, extent) characteristics correlate with the serum gastrin increase post secretin and post calcium. However, using the proposed criteria, the result of these provocative tests (that is, positive or negative) is minimally influenced by these factors, so secretin and calcium provocative tests are reliable in patients with different clinical, laboratory, and tumor characteristics. A systematic analysis of meal testing showed that 54%-77% of ZES patients have a <50% postprandial serum gastrin increase. However, 9%-20% of ZES patients had a >100% increase post meal, causing significant overlap with antral syndromes. Furthermore, we could not confirm the usefulness of meal tests for localization of duodenal gastrinomas. We conclude that the secretin test is a crucial element in the diagnosis of most ZES patients, the calcium test may be useful in selected patients, but the meal test is not helpful in the management of ZES. For secretin testing, the criterion with the highest sensitivity and specificity is an increase of > or =120 pg/mL, which should replace other criteria commonly used today. PMID- 17108780 TI - Sinonasal involvement in sarcoidosis: a case-control study of 20 patients. AB - We conducted a retrospective single-center study to describe the clinical features of sinonasal sarcoidosis (SNS) and to determine whether SNS is associated with a particular clinical phenotype of sarcoidosis. Twenty patients with histologically proven SNS (men/women, 7/13; mean age, 32 +/- 9 yr) were compared with control patients with sarcoid but without sinonasal (SN) involvement. Each patient was matched with 2 controls for the date of admittance in our institution. SN involvement occurred in the course of previously known sarcoidosis in 8 patients, whereas it preceded disease diagnosis in 12 patients. Among these 12 patients, 4 initially presented with strictly isolated SNS and 8 had other associated signs related to sarcoidosis. The most common symptoms were stuffiness (90%), anosmia (70%), and rhinorrhea (70%). Lupus pernio was frequent (50%). Local examination was constantly abnormal and showed hypertrophy (75%) and purplish coloring of the nasal mucosa with granulations (50%) on the septum and/or inferior turbinates. Computed tomography scans showed medial lytic lesions, mainly of the septum and/or the turbinates in about half the cases. All patients had negative antineutrophil cytoplasmic antibodies. Patients with SNS had significantly more frequent and severe involvement of vital organs than controls, had a longer history of sarcoidosis, and required systemic treatment more frequently (100% vs. 57.7%, p < 0.001) and for a longer time (78 +/- 42 mo vs. 29 +/- 18 mo, p < 0.0001). Corticosteroids maintenance dosage was high (10.5 +/- 6 mg daily) and mainly depended on SN involvement. Although rare, SN involvement is a severe and recalcitrant manifestation of sarcoidosis representing a therapeutic challenge. PMID- 17108781 TI - Addressing the underperformance of faculty and staff. AB - Many new nursing leaders assuming work as deans, assistant deans, or interim deans have limited education, experience, or background to prepare them for the job. To assist new deans and those aspiring to be deans, the authors of this department, both deans, offer survival tips based on their personal experiences and insights. They address common issues, challenges, and opportunities that face academic executive teams, such as negotiating an executive contract, obtaining faculty lines, building effective work teams, managing difficult employees, and creating nimble organizational structure to respond to changing consumer, healthcare delivery, and community needs. The authors welcome counterpoint discussions with readers. PMID- 17108782 TI - Supporting faculty proposal development and publication. AB - The Hispanic Health Disparities Research Center, a collaborative venture between the University of Texas at El Paso and the University of Texas at Houston Health Sciences Center, supports the research capabilities of junior faculty through a variety of programs. Novice researchers often need practical help in conducting literature reviews, extracting data, evaluating the evidence, and formulating a research question of significant importance to be funded yet narrow enough to fit within the scope of the proposal. The authors discuss a successful proposal development program that includes mentoring by more senior faculty and structured sessions with a medical writer and editor. PMID- 17108783 TI - The search for a major professor. PMID- 17108784 TI - Preparing a nursing library for National League for Nursing Accrediting Commission accreditation. PMID- 17108785 TI - Diversity BINGO: a strategy to increase awareness of diversity in the classroom. PMID- 17108786 TI - Educating international nurses: curricular innovation through a bachelor of science in nursing bridging program. AB - A curricular innovation was designed to provide internationally educated nurses with access to nursing licensure and employment. Through a program that includes professionally relevant English language support, mentorship, academic upgrading, workplace experiences, and clinical skills support, a mechanism has been created for internationally educated nurses to earn a bachelor of science in nursing degree and overcome barriers to practicing their profession. PMID- 17108787 TI - Giving and receiving: a case study in service learning. AB - Service learning is becoming a requirement for many universities wanting to connect with the communities surrounding them while also instilling civic responsibility in their students. The process should benefit all involved. In the following case study, the authors describe their experience of initiating a service learning component in an introductory nursing course, from the design to the implementation of the project. Faculty concerns and student reactions are included. PMID- 17108788 TI - Simulated clinical experience: nursing students' perceptions and the educators' role. AB - Simulation with high-fidelity technology is an innovative and effective teaching strategy to address increasing student enrollment, faculty shortages, and limited clinical sites. This interactive learning method allows the educator to focus on desired content and teach multiple objectives. This study examines students' perceptions of a preterm labor simulated clinical experience as a method of instruction, emphasizing the importance of the educators' role in promoting positive student outcomes. PMID- 17108789 TI - Nurse entrance test scores: a predictor of success. AB - A program evaluation was conducted to determine if requiring higher scores on critical thinking components of the Nurse Entrance Test would have a positive effect on the percentage of students that could be retained in a diploma nursing program. The program evaluation revealed that using the Nurse Entrance Test as a tool for admissions screening, specifically portions of the examination that predict critical thinking, was effective in helping to predict success through level I nursing courses. PMID- 17108790 TI - Using film clips to enhance nursing education. AB - Film clips in nursing education provide a unique way to promote active learning. Several methods for using film clips and combining them with other teaching strategies are discussed. Suggestions for the use of popular clips, logistical aspects of showing clips in class, and legal issues associated with copyright are addressed. Faculty are encouraged to use this strategy to evoke emotional responses, generate discussion, enhance clinical decision making, and provide vicarious learning. PMID- 17108792 TI - Strategies to manage the time demands of online teaching. AB - The time demands of online teaching may discourage nurse educators from developing distance learning courses. As consumer expectations for online education grow, it is essential that educators identify strategies that will enable efficient use of time to both develop and teach online courses. The authors review the distance learning literature from the perspective of time management strategies for online teaching. Approaches used by the authors to efficiently develop and teach online courses are described. PMID- 17108795 TI - The International Classification of Functioning, Disability and Health. PMID- 17108794 TI - Using qualitative methods for course evaluation: a case study from Botswana. AB - This article is a report of a qualitative evaluation of a course on human immunodeficiency virus/acquired immunodeficiency syndrome carried out jointly by faculty from Botswana and the United States at a university in Botswana. It demonstrates the importance of both international nurse educator expertise in impacting a major pandemic and the use of qualitative methods for course evaluation. PMID- 17108796 TI - Relationships among range of motion, functional mobility, and quality of life in children and adolescents after limb-sparing surgery for lower-extremity sarcoma. AB - PURPOSE: The study was designed to examine relationships between range of motion (ROM), functional mobility, and quality of life (QL) in patients with lower extremity sarcoma (LES) after limb-sparing surgery METHODS: Sixty-eight patients with LES (age, 10-26 years) participated. The patients performed hip flexion, hip extension, knee flexion, and knee extension, Timed Up and Down Stairs (TUDS), Timed Up and Go (TUG), nine-minute run-walk (9-min), and completed the QL measure, Short-Form-36 version two (SF-36v2). RESULTS: Significant correlations (p < 0.01) were found between hip extension and SF-36v2 physical component scale (PCS; r = 0.33), TUDS (r = -0.32), TUG (r = -0.33); hip flexion and TUDS (r = 0.31), TUG (r = -0.39), 9-min (r = 0.44); knee flexion and TUDS (r = -0.52), TUG (r = -0.40), 9-min (r = 0.37); SF-36v2 PCS and TUDS (r = -0.56), TUG (r = -0.51), 9-min (r = 0.60). CONCLUSION: ROM correlates with functional mobility and QL in patients with LES after limb-sparing surgery. ROM exercises are important component of a physical therapy program for children and adolescents with LES. PMID- 17108797 TI - The effects of therapeutic taping on gross motor function in children with cerebral palsy. AB - PURPOSE: Therapeutic taping to address dysfunctional sitting control in children with cerebral palsy (CP) was investigated in this study. METHODS: Eighteen children with quadriplegic CP, Gross Motor Function Classification System for Cerebral Palsy levels IV (n = 9) and V (n = 9) participated in the 12-week program. Subjects were assigned randomly to one of two groups: therapeutic taping + physical therapy or physical therapy only. Therapeutic taping was applied for periods of up to 72 hours over the paraspinal region. The effects were assessed with the Gross Motor Function Measure (GMFM-88) at baseline, six weeks, and 12 weeks. A factorial analysis of variance was used to examine group differences over time. RESULTS: No significant differences were found for the GMFM-88 scores between groups over time. CONCLUSION: Therapeutic taping does not evoke a positive functional change in the seated postural control of children with quadriplegic cerebral palsy. Subjective observation, however, suggested that one child with athetosis benefited from therapeutic taping over the paraspinal region. PMID- 17108798 TI - Relationships among measures of knee musculoskeletal impairments, gross motor function, and walking efficiency in children with cerebral palsy. AB - PURPOSE: The study was designed to determine whether relationships exist among knee muscle spasticity, isometric knee muscle strength, knee muscle balance, gross motor function, and walking efficiency in children with spastic cerebral palsy and to evaluate the relative contributions of impairment measures to functional outcome measures. METHODS: Twenty-seven children with spastic diplegic cerebral palsy participated. Knee muscle strength and spasticity were measured by a hand-held dynamometer and the Ashworth Scale, respectively. Functional abilities were assessed by the Gross Motor Function Measure and by calculating the energy expenditure index from data collected during two three-minute walk tests (comfortable and fast speeds). RESULTS: Significant relationships were found between impairments and functional abilities (r = 0.42-0.85, p < 0.05). Isometric hamstring strength and quadriceps spasticity explained 81.5% of total Gross Motor Function Measure score variance. Quadriceps spasticity explained 56.7% of energy expenditure index variance during comfortable walking. CONCLUSION: Hamstring strength and quadriceps spasticity explain much of the variance in gross motor function and comfortable walking efficiency. PMID- 17108799 TI - Within-session inter-rater realiability of goniometric measures in patients with spastic cerebral palsy. AB - PURPOSE: The purpose of this study was to investigate whether appropriate inter rater reliability of goniometric measurement could be achieved in children with cerebral palsy (CP) by a less-experienced and more-experienced therapist within the same session in a clinic setting. SUBJECTS: Twenty-five children with spastic CP underwent goniometric measurement of their right and/or left legs (46 legs). METHODS: Two pediatric physical therapists, with 10 and one year of experience measured five lower-extremity motions within the same session. Standard positioning, stabilization, and bony landmarks were reviewed before the study. RESULTS: Interclass correlation coefficients ranged from 0.582 (hip extension) to 0.929 (popliteal angle). CONCLUSION: Our data are in agreement with the literature on the reliability of range of motion measurements in children with CP. As a result, we were able to show that inter-rater goniometric reliability can be established between a less-experienced and more-experienced therapist within the same session. PMID- 17108800 TI - Increasing ankle strength to improve gait and function in children with cerebral palsy: a pilot study. AB - PURPOSE: This pilot study was designed to determine whether increases in ankle strength could improve gait and function in children with spastic diplegia. METHODS: Data were obtained from 12 children with spastic diplegia who were assigned randomly to a dorsiflexor group, a plantarflexor group, a dorsi- and plantarflexor group, or a control group. Training group subjects participated in a 12-week strength program using an isokinetic dynamometer. RESULTS: The majority of subjects increased their strength in the trained muscles; the strength of untrained muscles was unchanged. The Gross Motor Function Measure (GMFM) walk-run jump dimension and a quality of life measure (ie, Peds QL) improved significantly for the entire training group. Gait speed was not significantly increased for the entire training group but varied among training groups. Gait kinematics significantly improved or showed trends for improvement for the entire training group. CONCLUSION: This pilot investigation indicates that ankle strength increases may lead to improved function, gait speed, and quality of life in children with spastic diplegia. PMID- 17108801 TI - Use of a lumbosacral orthosis in the management of septic arthritis of the hip. AB - INTRODUCTION: Septic arthritis can lead to loss of joint motion and permanent mobility deficits because of infection and inflammatory response damaging articular structures. METHODS: A 13-year-old boy underwent five left hip surgeries after diagnosis of septic arthritis. He participated in 14 months of physical therapy, including hip stretching and manipulation. A lumbosacral orthosis (LSO) was applied to immobilize the lumbar region and isolate motion at the left hip. Treatment was divided into three phases: initial treatment, after manipulation under anesthesia, and after application of the LSO. Hip flexion range of motion (ROM) improved by 100 degrees and extension by 20 degrees. The patient returned to a high level of function, including athletics, but continued to have limited ROM and gait deviations. CONCLUSIONS: Improvement in hip ROM after application of the LSO suggests that this intervention may have applicability in treatment of hip dysfunctions. PMID- 17108802 TI - Adaptive downhill skiing in children with cerebral palsy: effect on gross motor function. AB - PURPOSE: The study was designed to examine the effect of adaptive downhill skiing (ADS) on gross motor function in children with spastic cerebral palsy. METHODS: One girl and four boys participated (mean age = eight years, five months). All were ambulatory. Participants' Gross Motor Function Measure (GMFM) classifications were: Level I (n = 2); Level II (n = 2); Level III (n = 1). ADS was conducted for a 10-week period at one ski resort. Each participant had the same ski instructor. GMFM was obtained every five weeks: beginning five weeks before ADS instruction and continuing to 10 weeks after ADS instruction. RESULTS: After 10 weeks of ADS GMFM-D, and GMFM-Total Score increased 5.4% (p = 0.022) and 3.2% (p = 0.035), respectively, and remained increased 10 weeks after ADS. CONCLUSION: ADS could be recommended by clinicians as a recreational activity for the gross motor rehabilitation of ambulatory children with spastic cerebral palsy. PMID- 17108805 TI - How do we treat pediatric patients with spastic diplegia? PMID- 17108808 TI - Adding pharmacogenetics information to drug labels: lessons learned. AB - The US Food and Drug Administration approved a revised package insert for two cancer drugs to include information about the increased risk of severe adverse events owing to enzyme deficiencies caused by genetic variants. The label revisions stopped short of recommending or requiring pharmacogenetic testing prior to or following an adverse event. Despite (or because of) the lack of specific recommendations, we believe the actions taken by US Food and Drug Administration will have implications for pharmacogenetics research, clinical integration, and other policy considerations. We review the reasons behind the cautious label changes and discuss some of the lessons that can be learned from these experiences. PMID- 17108809 TI - In-vitro transport characteristics discriminate wild-type ABCB1 (MDR1) from ALA893SER and ALA893THR polymorphisms. AB - BACKGROUND: Genetic variation of the human ABCB1 (P-glycoprotein; MDR1 gene product) efflux transporter is strongly suggested as a determinant factor governing the pharmacokinetics of diverse drugs and xenobiotics. Despite various efforts to associate polymorphisms in ABCB1 to actual clinical effect and transport function, information is still inconsistent or even controversial. METHODS AND RESULTS: Using membrane vesicle preparation from ABCB1-expressing HighFive insect cells, we report here that saturation kinectic parameters of the frequently occurring ABCB1 triallelic variants 893Ser (exon 21, 2677T) and 893Thr (2677A) were considerably different from wild-type 893Ala (2677G), despite similar protein expression levels. Of importance were significant differences in transport capacities between the tested 893Ala/Ser/Thr variants. In comparison with 893Ala, maximal transport rates for vincristine of 893Ser and 893Thr increased 50% and three-fold, respectively. Cis-inhibition by digoxin, didanosine or fexofenadine was least pronounced in 893Ser, whereas no genotype differences could be observed using verapamil. CONCLUSION: These results suggest an influence of ABCB1-893 triallelic variants on transport function and drug-drug interaction, which might be most pronounced in 893Thr. Furthermore, some of the mechanisms of 2677G/T/A-based haplotype-associated alterations in ABCB1 activity may have been unveiled. PMID- 17108810 TI - Arsenic-related skin lesions and glutathione S-transferase P1 A1578G (Ile105Val) polymorphism in two ethnic clans exposed to indoor combustion of high arsenic coal in one village. AB - OBJECTIVES: A total of 2402 patients with arsenic-related skin lesions, such as hyperkeratosis, hyperpigmentation or hypopigmentation, or even skin cancer in a few villages in Southwest Guizhou Autonomous Prefecture, China represent a unique case of endemic arsenism related with indoor combustion of high arsenic coal. This study aimed to investigate the cluster of arsenism cases and the possible relevant factors including GSTP1 polymorphism in two clans of different ethnic origin living in one village for generations. METHODS: A questionnaire-based study was performed in 170 Miao clan P members, 10 of whom had arsenic-related skin diseases, and 153 Han clan G1 members, 50 of whom had arsenic-related skin diseases. The data were checked against the registration archives since the 1980s. At the same time, arsenic concentrations in samples of coal, indoor air, drinking water, corn and chilli pepper that were once baked over the stoves for desiccation, as well as in samples of urine and hair of clan members were determined. Glutathione S-transferase P1 (GSTP1) A1578G polymorphism was genotyped by a restriction fragment length polymorphism-based procedure. RESULTS: Arsenism morbidity in Miao clan P was significantly lower than in the neighbouring Han clan G1 [5.9 vs. 32.7%, odds ratio (OR)=0.13, 95% confidence interval (CI): 0.06-0.27, P<0.0001]. No sex differences were confirmed inside both clans. Analyses of the environmental samples indicated that Miao clan P members were exposed to higher amounts of arsenic via inhalation and food ingestion. Hair and urine samples also proved a higher arsenic body burden in ethnic Miao individuals. No corresponding differences by sex were found. Higher frequencies of combined mutant genotype G/G1578 and A/G1578 (OR=4.72, 95% CI: 2.34-9.54, P<0.0001) and of mutant allele G1578 (OR=3.22, 95% CI: 2.00-5.18, P<0.0001) were detected in diagnosed arsenism patients than in non-diseased individuals. The Miao individuals showed a lower percentage of combined mutant genotypes (30.6 vs. 52.7%, OR=0.40, 95% CI: 0.19-0.84, P=0.015) as well as of mutant allele G1578 (OR=0.46, 95% CI: 0.24-0.88, P=0.017) than their Han neighbours. CONCLUSIONS: Genetic predisposition influences dermal arsenism toxicity. The GSTP1 A1578G (Ile105Val) status might be a susceptibility factor for arsenic-related skin lesions. PMID- 17108811 TI - SLCO1B1 polymorphism markedly affects the pharmacokinetics of simvastatin acid. AB - BACKGROUND AND OBJECTIVE: Organic anion transporting polypeptide 1B1 (OATP1B1) is an uptake transporter located at the sinusoidal membrane of human hepatocytes. This study aimed to investigate the effects of genetic polymorphism in the SLCO1B1 gene encoding OATP1B1 on the pharmacokinetics of simvastatin. METHODS: Four healthy volunteers with the homozygous SLCO1B1 c.521CC genotype, 12 with the heterozygous c.521TC genotype and 16 with the homozygous c.521TT genotype (controls) were recruited. Each study participant ingested a single 40-mg dose of simvastatin. Plasma concentrations of simvastatin (inactive lactone) and its active metabolite simvastatin acid were measured for 12 h. RESULTS: The AUC0 infinity of simvastatin acid was 120 and 221% higher in participants with the SLCO1B1 c.521CC genotype than in those with the c.521TC and c.521TT (reference) genotypes, respectively (P<0.001). The Cmax of simvastatin acid was 162 and 200% higher in participants with the c.521CC genotype than in those with the c.521TC and c.521TT genotypes (P<0.001). The Cmax of simvastatin acid occurred earlier in participants with the c.521CC and c.521TC genotypes than in those with the c.521TT genotype (P<0.05). No association existed between the SLCO1B1 genotype and the elimination half-life of simvastatin acid. Moreover, no statistically significant association was seen between the SLCO1B1 genotype and the pharmacokinetics of simvastatin lactone. CONCLUSIONS: SLCO1B1 polymorphism markedly affects the pharmacokinetics of active simvastatin acid, but has no significant effect on parent simvastatin. Raised plasma concentrations of simvastatin acid in patients carrying the SLCO1B1 c.521C variant allele may enhance the risk of systemic adverse effects during simvastatin treatment. In addition, reduced uptake of simvastatin acid by OATP1B1 into the liver in patients with the c.521C allele could reduce its cholesterol-lowering efficacy. PMID- 17108812 TI - Liver X receptor gene polymorphisms and adipose tissue expression levels in obesity. AB - OBJECTIVE: LXRA and LXRB genes regulate adiposity, energy dissipation, as well as glucose and lipid homeostasis in mice. We investigated the LXR genes in human obesity. METHODS: LXRA and LXRB mRNAs were quantified in abdominal subcutaneous adipose tissue of obese and nonobese women. The LXRA and LXRB genes were screened for polymorphisms and common single nucleotide polymorphisms genotyped in obese and nonobese women. RESULTS: Relative LXRA mRNA expression levels were higher in obese women (P=0.03). One LXRA single nucleotide polymorphism, rs2279238, and one common haplotype, CAAGCC, as well as two LXRB single nucleotide polymorphisms, LB44732G>A and rs2695121, were associated with obesity phenotypes (nominal P values of 0.0075, 0.0014, 0.008 and 0.02, respectively). Furthermore, there was evidence of interaction between LXRA and LXRB alleles in determining body mass index. CONCLUSION: Our results support a role for LXRA in human adipose tissue. The nominal associations of LXRA and LXRB alleles with obesity are interesting and should be further investigated in independent data sets. PMID- 17108813 TI - NOS3 polymorphisms, cigarette smoking, and cardiovascular disease risk: the Atherosclerosis Risk in Communities study. AB - OBJECTIVE: Endothelial nitric oxide synthase (NOS3) activity and cigarette smoking significantly influence endothelial function. We sought to determine whether cigarette smoking modified the association between NOS3 polymorphisms and risk of coronary heart disease or stroke. METHODS: All 1085 incident coronary heart disease cases, all 300 incident ischemic stroke cases, and 1065 reference individuals from the Atherosclerosis Risk in Communities study were genotyped for the T-786C and E298D polymorphisms in NOS3. Using a case-cohort design, associations between genotype/haplotype and disease risk were evaluated by multivariable proportional hazards regression. Multiplicative scale interaction testing evaluated the influence of cigarette smoking history at baseline on these associations. RESULTS: In Caucasians, association between E298D genotype and risk of coronary heart disease was significantly modified by current smoking status (interaction P=0.013), with the highest risk observed in smokers carrying the variant D298 allele relative to nonsmokers carrying two E298 alleles (adjusted hazard rate ratio 2.07, 95% confidence interval 1.39-3.07). In African-Americans, association between T-786C genotype and risk of ischemic stroke was significantly modified by pack-year smoking history (interaction P=0.037), with the highest risk observed in >or=20 pack-year smokers carrying the variant C-786 allele relative to <20 pack-year smokers carrying two T-786 alleles (adjusted hazard rate ratio 4.03, 95% confidence interval 1.54-10.6). CONCLUSIONS: An interaction between the E298D and T-786C polymorphisms in NOS3, cigarette smoking, and risk of incident coronary heart disease and ischemic stroke events appears to exist, suggesting a potential complex interplay between genetic and environmental factors and cardiovascular disease risk. PMID- 17108814 TI - DRD2 genetic variation in relation to smoking and obesity in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. AB - OBJECTIVES: Cigarette smoking is the leading cause of morbidity and mortality worldwide. We investigated the association between smoking behavior and genetic variations in the D2 dopamine receptor (DRD2), which mediates nicotine dependence. To assess the specificity of genetic effects, we also investigated other reward-motivated characteristics (obesity, alcohol consumption). METHODS: Four single nucleotide polymorphisms in DRD2 were genotyped in 2374 participants selected randomly from the screening arm of the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial after stratifying by sex, age, and smoking status. Smoking, obesity, and alcohol consumption were assessed by questionnaire. Single nucleotide polymorphism and haplotype associations were estimated using odds ratios (ORs) and 95% confidence intervals derived from conditional logistic regression models, adjusted for race/ethnicity. RESULTS: DRD2 polymorphisms were associated with the risk of remaining a current smoker and obesity. Current smokers were more likely than former smokers to possess the variant TaqIA allele (rsmusical sharp1800497) in a dose-dependent model (ORCT=1.2, ORTT=1.5, P for linear trend=0.007). The DRD2 haplotype T-C-T-A [TaqIA(C/T)-957(T/C)-IVS6-83(G/T) -50977(A/G)] was more common among current than former smokers (OR=1.3, P=0.006), particularly among heavy smokers (21+ cigarettes per day; OR=1.6, P=0.006), and was more common among obese than normal weight individuals (OR=1.4, P=0.02). CONCLUSIONS: Genetic variation in DRD2 is a modifier of the reward motivated characteristics, smoking and obesity. As fewer than 15% of smokers who attempt to quit are able to maintain abstinence for greater than 3 months, our results support that DRD2 is an appropriate molecular target for smoking cessation treatments. Our results further support evaluation of DRD2 antagonists for obesity therapies. PMID- 17108815 TI - Polymorphism in IgG Fc receptor gene FCGR3A and response to infliximab in Crohn's disease: a subanalysis of the ACCENT I study. AB - Recently, it has been shown that FCGR3A-158 gene polymorphism is associated with biological and possibly clinical response to infliximab in Crohn's disease. We further assessed this association in a subset of 344 patients from the large and well-defined cohort of 573 patients with Crohn's disease from the ACCENT I study. No association could be observed between FCGR3A-158 gene polymorphism and the clinical response to infliximab, which was primarily defined as a decrease of >or=70 points in the Crohn's disease activity index or clinical remission (Crohn's disease activity index <150). We did, however, confirm a trend towards a greater decrease in C-reactive protein after infliximab in V/V homozygotes as compared with V/F heterozygotes and F/F homozygotes (-79.4, -76.5, and -64.3%, respectively, at week 6; P=0.085; one-tailed P=0.043). This finding has no immediate clinical impact but may enhance the understanding of the complex mechanisms of action of anti-tumor necrosis factor agents in Crohn's disease. PMID- 17108817 TI - Transforaminal posterolateral endoscopic discectomy with or without the combination of a low-dose chymopapain: a prospective randomized study in 280 consecutive cases. AB - STUDY DESIGN: A prospective randomized study involving 280 consecutive cases of lumbar disc herniation managed either by an endoscopic discectomy alone or an endoscopic discectomy combined with an intradiscal injection of a low dose (1000 U) of chymopapain. OBJECTIVE: To compare outcome, complications, and reherniations of both techniques. SUMMARY OF BACKGROUND DATA: Despite a low complication rate, posterolateral endoscopic nucleotomy has made a lengthy evolution because of an assumed limited indication. Chemonucleolysis, however, proven to be safe and effective, has not continued to be accepted by the majority in the spinal community as microdiscectomy is considered to be more reliable. METHOD: A total of 280 consecutive patients with a primary herniated, including sequestrated, lumbar disc with predominant leg pain, was randomized. A clinical follow-up was performed at 3 months, and at 1 and 2 years after the index operation with an extensive questionnaire, including the visual analog scale for pain and the MacNab criteria. The cohort integrity at 3 months was 100%, at 1 year 96%, and at 2 years 92%. RESULTS: At the 3-month evaluation, only minor complications were registered. At 1-year postoperatively, group 1 (endoscopy alone) had a recurrence rate of 6.9% compared to group 2 (the combination therapy), with a recurrence rate of 1.6%, which was a statistically significant difference in favor of the combination therapy (P = 0045). At the 2-year follow up, group 1 reported that 85.4% had an excellent or good result, 6.9% a fair result, and 7.7% were not satisfied. At the 2-year follow-up, group 2 reported that 93.3% had an excellent or good result, 2.5% a fair result, and 4.2% were not satisfied. This outcome was statistically significant in favor of the group including chymopapain. There were no infections or patients with any form of permanent iatrogenic nerve damage, and no patients had a major complication. CONCLUSIONS: A high percentage of patient satisfaction could be obtained with a posterior lateral endoscopic discectomy for lumbar disc herniation, and a statistically significant improvement of the results was obtained when an intradiscal injection of 1000 U of chymopapain was added. There was a low recurrence rate with no major complications. The method can be applied in any type of lumbar disc herniation, including the L5-S1 level. PMID- 17108818 TI - A kinematic model to assess spinal motion during walking. AB - STUDY DESIGN AND OBJECTIVES: A 3-dimensional multi-segment kinematic spine model was developed for noninvasive analysis of spinal motion during walking. Preliminary data from able-bodied ambulators were collected and analyzed using the model. SUMMARY OF BACKGROUND DATA: Neither the spine's role during walking nor the effect of surgical spinal stabilization on gait is fully understood. Typically, gait analysis models disregard the spine entirely or regard it as a single rigid structure. Data on regional spinal movements, in conjunction with lower limb data, associated with walking are scarce. METHODS: KinTrak software (Motion Analysis Corp., Santa Rosa, CA) was used to create a biomechanical model for analysis of 3-dimensional regional spinal movements. Measuring known angles from a mechanical model and comparing them to the calculated angles validated the kinematic model. Spine motion data were collected from 10 able-bodied adults walking at 5 self-selected speeds. These results were compared to data reported in the literature. RESULTS: The uniaxial angles measured on the mechanical model were within 5 degrees of the calculated kinematic model angles, and the coupled angles were within 2 degrees. Regional spine kinematics from able-bodied subjects calculated with this model compared well to data reported by other authors. CONCLUSIONS: A multi-segment kinematic spine model has been developed and validated for analysis of spinal motion during walking. By understanding the spine's role during ambulation and the cause-and-effect relationship between spine motion and lower limb motion, preoperative planning may be augmented to restore normal alignment and balance with minimal negative effects on walking. PMID- 17108819 TI - Computed tomography evaluation of spondylolysis and spondylolisthesis in asymptomatic patients. AB - STUDY DESIGN: A retrospective radiographic study involving analysis of abdominal and pelvic computed tomography (CT) scans obtained on patients presenting with clinical conditions other than back pain. OBJECTIVE: To determine the incidence of spondylolysis and spondylolisthesis in patients requiring inpatient or emergency department CT evaluation for unrelated abdominal and pelvic conditions. SUMMARY OF BACKGROUND DATA: Spondylolysis and spondylolisthesis are part of a disease process that is thought to be resultant from biomechanical stresses related to bipedal locomotion. The incidence is estimated to be 3% to 10% in the general population. Many of these cases occur without associated symptoms. To our knowledge, there is a relative paucity of data on the use of CT to evaluate the prevalence of these 2 entities in patients seeking medical attention for unrelated conditions. METHODS: Five hundred ten consecutive abdominal and pelvic multi-detector CT scans obtained on a single scanner (Philips MX8000; Eindhoven, The Netherlands) were reviewed. These patients presented with such complaints as abdominal pain and fever, or were imaged as part of their inpatient evaluation for conditions unrelated to lumbar spine pathology. A board certified radiologist and a radiology resident retrospectively evaluated CT scans for lumbar spondylolysis, spondylolisthesis, and associated degenerative changes. A neuroradiologist confirmed all positive cases. RESULTS: Of the 510 cases examined, there were 29 cases of spondylolysis at L5, corresponding to a prevalence of 5.7%. Twenty-three of the cases demonstrated bilateral spondylolysis and 6 unilateral. Sixteen of the 23 cases of bilateral spondylolysis also had spondylolisthesis, 13 of which were grade I, and 3 of which were grade II. In patients 45 years old and younger who did not have spondylolysis or spondylolisthesis, we observed a 32.2% incidence of sclerosis involving the L5 lumbar pedicles. CONCLUSIONS: This study demonstrates a 5.7% prevalence of spondylolysis and a 3.1% prevalence of spondylolisthesis in patients undergoing CT scans of the abdomen and pelvis for unrelated reasons, corresponding to the rate of spondylolysis and spondylolisthesis detected in prospective plain radiographic studies. We observed a 1.2% incidence of unilateral spondylolysis, and approximately 67% of these demonstrated contralateral sclerosis. It is suggested in the literature that sclerosis of the contralateral pedicle seen in cases of unilateral spondylolysis may be a compensatory response to mechanical stresses on an unstable lumbar vertebral body. PMID- 17108820 TI - Electromagnetic topographical technique of curve evaluation for adolescent idiopathic scoliosis. AB - STUDY DESIGN: Diagnostic testing. OBJECTIVE: The goal of this study is to measure the accuracy and reliability of the Orthoscan (Orthoscan Technologies, Inc.) and to determine whether it can be substituted for radiographs in the surveillance of adolescent idiopathic scoliosis (AIS). SUMMARY OF BACKGROUND DATA: AIS is usually followed using scoliosis radiographs, which offer the most reliable way to quantify the curve, but carry the risk of exposure to ionizing radiation. The Orthoscan is a nonradiographic topographic method for measuring spinal curves. MATERIALS AND METHODS: There were 5 phases of this study that measured: the accuracy and reliability of the machine when used with a plastic model; the variability with a real patient; the intraobserver variability; the correlation between the measurements of the machine and that of the radiograph; and the correlation between the change in radiograph measurement over time and the change in Orthoscan measurement over time. RESULTS: In measurement of a static plastic model, the machine measured curves with a standard deviation of +/-1 degrees in trunk rotation and +/-2 degrees in curve measurement. Error increased with a real patient. Thirty-six comparisons in the thoracic spine, and 19 comparisons in the lumbar spine, were made between measurements using the Orthoscan and radiographs. Mean curves in the 2 groups were not significantly different and had poor-to moderate correlation. Longitudinal evaluation included 47 curves in 28 patients. The Orthoscan predicted the radiograph change within an acceptable range 55.3% of the time. CONCLUSIONS: The Orthoscan does not accurately predict the scoliosis curve magnitude or the overall change in curve over time. While analysis in groups of patients using this technique reveals group means that begin to look acceptable, if the variability is too great, then this technology is not yet ready to replace the radiograph in the evaluation of a scoliosis curve. PMID- 17108822 TI - A cemented caval vein filter: case report. AB - STUDY DESIGN: Case report. OBJECTIVE: To present a previously undescribed complication of bone cement use during spinal surgery. SUMMARY OF BACKGROUND DATA: Surgical cement reaching the circulation is a frequently described complication. The placement of caval vein filters is a routine measure in specific patients to prevent thrombotic material, usually from deep venous thrombosis in the lower extremities, from reaching the pulmonary circulation. A case of bone cement getting trapped in the caval filter rendering it unremovable has not been published. METHODS: A 66-year-old patient underwent dorsal spondylodesis of the lumbar spine for superinfected metastasis with instability. Because of deep venous thrombosis of both femoral and iliac veins, a caval filter had been placed before surgery. Bone cement from the surgical procedure migrated into the venous bloodstream and got caught in the caval filter, thus rendering the filter unremovable. RESULTS: Surgical removal of the filter was necessary. CONCLUSION: If caval filters are present in patients undergoing surgery involving the use of bone cement, the possibility of cement being caught by the filter must be considered. PMID- 17108823 TI - Spinal cord compression in a patient with multiple hereditary exostoses caused by breast adenocarcinoma metastatic to osteochondromas of the spine: case report. AB - STUDY DESIGN: Case report. OBJECTIVE: To report on thoracic spinal cord compression caused by a mass in a 66-year-old female with new onset of myelopathic symptoms and a history of multiple hereditary exostoses. SUMMARY OF BACKGROUND DATA: To our knowledge, there have been no previous reports of spinal cord compression in a patient with multiple hereditary exostoses caused by breast adenocarcinoma metastatic to osteochondromas of the spine. METHODS.: Chart, pathologic, and radiographic documentation of the preoperative and postoperative clinical course of the patient was used. RESULTS: The patient had resolution of her neurologic symptoms following wide surgical excision, decompression, and stabilization from T2 to T10. The patient's mass was found to be breast adenocarcinoma metastatic to osteochondromas of the spine. CONCLUSIONS: When faced with a patient with a history of multiple hereditary exostoses with new onset of myelopathic symptoms and a mass compressing the spinal cord, the clinician's differential should be broad and always initially include a metastatic lesion, osteochondroma, or chondrosarcoma. PMID- 17108824 TI - Transoral vertebroplasty for renal cell metastasis involving the axis: case report. AB - STUDY DESIGN: Case report. OBJECTIVE: This is one of the first reported cases of transoral vertebroplasty for a solid metastatic tumor at C2 body. SUMMARY OF BACKGROUND DATA: Percutaneous vertebroplasty has gained popularity as a treatment option for painful neoplastic lesions of the spine. The technique has been useful in reducing pain and stabilizing the spinal segment that is vulnerable to fracture. However, there is very little experience with vertebroplasty in the cervical spine and, more specifically, at C2. METHODS: We present a case of metastatic renal cell carcinoma of the C2 body. The patient, a 61-year-old woman, presented with an excruciating neck pain. Diagnostic workup revealed the presence of tumor in the right kidney. Multiple spinal levels were involved in addition to C2, and the C2 lesion was treated for palliative purpose. Under biplanar fluoroscopy, a vertebroplasty trocar was placed transorally into the central portion of the C2 body, and polymethyl methacrylate was injected. RESULTS: The transoral vertebroplasty achieved complete pain relief and enhanced stability of an extensive osteolytic lesion involving the C2 body. CONCLUSIONS: This minimal access procedure was effective in completely relieving pain from a metastatic deposit at C2, while adequately stabilizing the vulnerable segment by the injection of polymethyl methacrylate. The transoral route requires meticulous fluoroscopic control to prevent the leakage of polymethyl methacrylate but provides the most direct access to the C2 body. PMID- 17108825 TI - The treatment of disc-herniation-induced sciatica with infliximab: one-year follow-up results of FIRST II, a randomized controlled trial. AB - STUDY DESIGN: A randomized controlled trial. OBJECTIVES: To evaluate the long term efficacy of infliximab, a monoclonal antibody against tumor necrosis factor alpha (TNF-alpha), in patients with acute/subacute sciatica secondary to herniated disc. SUMMARY OF BACKGROUND DATA: The results of experimental studies and our open-label trial support the use of infliximab in sciatica. Here we report the 1-year results of a randomized controlled trial (FIRST II, Finnish Infliximab Related STudy) evaluating the efficacy and safety of a single infusion of infliximab for sciatic pain. METHODS: Inclusion criteria were unilateral sciatic pain with a disc herniation concordant with the symptoms and signs of radicular pain. Patients had to be candidates for discectomy. Criteria for discectomy included (in addition to a symptomatic disc herniation on MRI) neural entrapment (straight leg raising [SLR] < or =60 degrees ) with either a short term (2-4 weeks) severe or long-term (4-12 weeks) moderate leg pain. Forty patients were allocated to a single intravenous infusion of either infliximab 5 mg/kg or placebo. Differences in the clinical examination parameters (straight leg raise [SLR], muscle strength, sensory defects, tendon reflexes), patient reported symptoms (leg and back pain using a visual analog scale [VAS], Oswestry disability, quality-of-life [RAND-36]), sick leaves, number of discectomies, and adverse effects between the two treatment groups over the 1-year follow-up were compared using Mann-Whitney U test or Student's t test, repeated-measures analysis, or Cox proportional hazards model. Logistic regression was used to assess the predictors of good response. RESULTS: Sixty-seven percent of patients in the infliximab group reported no pain at 52 weeks compared with 63% in the control group (P = 0.72). Similar efficacy was observed between treatment groups for other outcomes. Eight patients in each group required surgery. Three nonserious adverse reactions were encountered in the infliximab group. The response (irrespective of the treatment) was significantly better with shorter symptom duration and less SLR restriction at baseline. Patients in the infliximab group appeared to especially benefit in cases of a L4-L5 (or L3-L4) herniation and if a Modic change was colocalized at the symptomatic level. CONCLUSIONS: Although the long-term results of this randomized trial do not support the use of infliximab compared with placebo for lumbar radicular pain in patients with disc herniation-induced sciatica, further study in a subgroup of patients with L4-L5 or L3-L4 herniations, especially in the presence of Modic changes, appears to be warranted. PMID- 17108826 TI - Anatomic considerations for the placement of C2 laminar screws. AB - STUDY DESIGN: Cadaveric study. OBJECTIVE: To study the applicability of C2 laminar screw placement in the general adult population and to provide useful guidelines for their safe placement. SUMMARY OF BACKGROUND DATA: Laminar screws for fixation into the second cervical vertebra are becoming an increasingly used technique since they eliminate the risk of vertebral artery injury. Although it is being used clinically, there are no published data that describe the anatomic considerations and potential limitations of this technique in the general population. METHODS: The C2 vertebrae of 420 adult specimens were studied. Laminar thickness, spinolaminar angle, and the length from the spinolaminar junction to the contralateral lamina/lateral mass junction were measured. Statistical analysis was performed using unpaired Student t tests and regression analysis (P < 0.05). RESULTS: Mean laminar thickness was 5.77 +/- 1.31 mm; 70.6% of specimens had a laminar thickness > or =5 mm; 92.6% had a thickness > or =4.0 mm. The spinolaminar angle was 48.59 degrees +/- 5.42 degrees. The mean screw length that could be used was 2.46 +/- 0.23 cm. More than 99% of specimens had an estimated screw length of at least 20 mm. Gender had a significant effect on all of the measurements studied, but race, height, and weight did not. CONCLUSION: The majority of specimens can safely accept placement of a laminar screw. This study establishes anatomic guidelines to allow for accurate screw selection and insertion. Preoperative planning is essential for safe screw placement via this technique. PMID- 17108827 TI - High rate of fusion in sheep cervical spines following anterior interbody surgery with absorbable and nonabsorbable implant devices. AB - STUDY DESIGN: Fourteen sheep were fused using anterior interbody implants at C2 C3 and C4-C5 and followed for 6 months. OBJECTIVE: To evaluate the effect of absorbable and nonabsorbable implants on fusion rate, cage migration, and implant integrity. SUMMARY OF BACKGROUND DATA: Despite the high clinical success rate with metallic plates and interbody grafting, complications such as dysphagia, imaging artifacts, and revision difficulties exist. Less permanent and lower profile implants could minimize these problems. METHODS: Four treatments were studied: 1) carbon fiber-reinforced polymer (CFRP) cage alone, 2) CFRP cage with an absorbable tension band, 3) absorbable cage with an absorbable tension band, and 4) absorbable cage with a titanium plate. Fusion was assessed using radiographs, biomechanical testing, and micro-CT analysis. RESULTS: Treatments with the CFRP cage alone had the lowest fusion rate at 3 months (2/6) and 6 months (4/6). The CFRP cage with the absorbable strap treatments had 5/6 fusions at 6 months. The absorbable cage with absorbable strap also had 5/6 fusions, but two of the cages fractured. The absorbable cage with titanium plate had 5/6 fusions, but the one partial fusion was attributed to poor screw and plate placement. CONCLUSION: Using both absorbable and nonabsorbable implants, high fusion rates were achieved in the challenging sheep cervical spine model. However, the absorbable cages were not able to withstand the mechanical forces during the 6-month survival period. PMID- 17108828 TI - B1 and TRPV-1 receptor genes and their relationship to hyperalgesia following spinal cord injury. AB - STUDY DESIGN: Laboratory investigation of pain behavior following spinal cord injury. OBJECTIVE: To explore changes in the spinal cord expression of nociceptive genes following spinal cord injury (SCI) as they relate to the manifestation of pain behavior in rats. SUMMARY OF BACKGROUND DATA: Neuropathic pain following SCI is common, disabling, and largely untreatable. In peripheral nerve injury models, bradykinin B1 and vanilloid 1 (TRPV-1) receptor activity is associated with neuropathic pain behavior. We sought to examine the role of these gene products in SCI-mediated pain. METHODS: Rats were subjected to SCI using the MASCIS impactor. Animals were tested preinjury and at regular intervals postinjury for the appearance of thermal hyperalgesia using a hind limb withdrawal latency test. The expression of B1 and TRPV-1 genes was assessed using real-time polymerase chain reaction. Immunohistochemistry was used to localize the B1 and TRPV-1 receptors within the spinal cord. RESULTS: Greater than twofold increases in the expression of the B1 and TRPV-1 genes were detected in the injured region of the spinal cord in animals exhibiting hyperalgesia compared with animals with SCI that did not display hyperalgesia. Immunohistochemical staining revealed that both receptor types were largely localized to the dorsal horn. Staining for TRPV-1 receptors decreased while that for B1 receptors increased in all of the injured animals when compared with sham-operated controls. CONCLUSION: B1 and TRPV-1 receptor genes are overexpressed in the injured spinal cord of animals manifesting thermal hyperalgesia following SCI compared with similarly injured animals without hyperalgesia. This finding is consistent with past work regarding the role of these receptors in nociception and indicates that ongoing modifiable processes are occurring in the spinal cord that lead to clinical pain syndromes. PMID- 17108829 TI - Anisotropic ion diffusivity in intervertebral disc: an electrical conductivity approach. AB - STUDY DESIGN: Investigation of the transport behavior of ions in intervertebral disc using an electrical conductivity method. OBJECTIVES: To determine the electrical conductivity and ion diffusivity of nucleus pulposus and anulus fibrosus in 3 major directions (axial, circumferential, and radial). SUMMARY OF BACKGROUND DATA: Knowledge of diffusivity of small molecules is important for understanding nutrition supply in intervertebral disc and disc degeneration. However, little is known on the anisotropic behaviors of ion diffusivity and of electrical conductivity in intervertebral disc. METHODS: Electrical conductivity measurement was performed on 24 axial, circumferential, and radial anulus fibrosus specimens and 24 axial nucleus pulposus specimens from bovine coccygeal discs. The diffusivity of Na and Cl were estimated by the analysis of conductivity data. RESULTS: The electrical conductivity (mean +/- standard deviation; n = 24) of the bovine anulus fibrosus was 4.70 +/- 1.08 mS/cm in the axial, 2.86 +/- 0.83 mS/cm in the radial, and 4.38 +/- 1.25 mS/cm in the circumferential direction. For nucleus pulposus, the electrical conductivity (mean +/- standard deviation; n = 24) was 8.95 +/- 0.89 mS/cm. The mean value for nucleus pulposus was significantly higher than that of anulus fibrosus (t test, P < 0.05). For anulus fibrosus, the conductivity in the radial direction was significantly lower than in axial or circumferential directions. Similar trends were found for both Na and Cl diffusivities. Both electrical conductivity and ion diffusivity were highly sensitive to water content. CONCLUSIONS: Electrical conductivity and ion diffusivity of anulus fibrosus are anisotropic. PMID- 17108830 TI - Biomechanical evaluation of a new total posterior-element replacement system. AB - STUDY DESIGN: In vitro study to characterize the flexibility of a new total posterior-element system when instrumented to L4-L5 segments. OBJECTIVE: The goal of this in vitro study was to investigate whether an optimized version of the TOPS implant (Impliant Ltd., Ramat Poleg, Israel) is capable to restore the physiologic motion characteristic of a spinal segment after facetectomy. SUMMARY OF BACKGROUND DATA: The TOPS implant is designed to replace the posterior elements of a functional spinal unit, to provide flexible restabilization and spinal alignment, while maintaining the intervertebral disc. The implant is composed of bilateral pedicle screws, connected with 2 crossbars in the transversal plane. The crossbars are joined together by an elastic element capable of transmitting tensile and compressive loads, as well as shear forces. METHODS: Six human cadaver specimens (L3-S1) (median age 61 years: minimum 47 and maximum 74 years) were used for this in vitro experiment. The specimens were loaded with pure moments of +/-7.5 Nm in flexion/extension, lateral bending, and axial rotation. The following states were investigated: (1) intact; (2) after bilateral laminectomy, including facetectomy of the lower facet joints, of the upper vertebra L4; and (3) after device implantation. The range of motion (ROM), neutral zone, and intradiscal pressure were determined from a third cycle. In a second step, the ROM in axial rotation was determined as a function of different flexion/extension postures. RESULTS: In the neutral position, the laminectomy and facetectomy increased the median values of the ROM in flexion plus extension, lateral bending right plus left, and significantly in axial rotation left plus right from: 8.2 degrees, 7.6 degrees, 3.6 degrees to 12.1 degrees, 8.5 degrees, and 8.5 degrees (Wilcoxon signed rank test; P < 0.05). After fixation of the implant, the ROM was again reduced to 6.8 degrees, 7.8 degrees, and 3.8 degrees. In a flexed posture, the ROM in axial rotation was slightly increased compared to the neutral position. With increasing extension, the axial rotation decreased linearly from 3.7 degrees in neutral position to 2.3 degrees in 4 degrees extension in the segment L4-L5. The characteristic of the intradiscal pressure versus load with the implant was similar to that of the intact specimen. CONCLUSION: The TOPS implant almost ideally restored the ROM in lateral bending and axial rotation compared to that of the intact specimen. In the sagittal plane, 85% of the intact ROM could be obtained. The ROM in axial rotation as a function of flexion and extension angle also mimics the biomechanical behavior of the posterior complex of a lumbar spine. This relationship between ROM and posture emphasizes the importance of a proper implantation. PMID- 17108832 TI - The feasibility of inserting atlas lateral mass screws via the posterior arch. AB - STUDY DESIGN: A total of 709 skeletally mature atlas specimens were obtained from the Hamann Todd Collection at the Cleveland Museum of Natural History. Using digital caliper, the thickness of the vertebral artery groove was measured to determine the feasibility of potential screw placement. OBJECTIVE: To determine the feasibility of screw insertion into the atlas lateral mass via the posterior arch. SUMMARY OF BACKGROUND DATA: The originally described starting point for atlas lateral mass screws, at the base of the lateral mass, can be associated with excessive bleeding and irritation of the greater occipital nerve. In order to avoid such problems, we routinely start the screw at a more cranial location on the dorsal, posterior-lateral arch of the atlas, when the anatomy permits. In cases where the arch is too small, we notch the undersurface of the arch so as to recess the screw into the notch. We undertook this cadaveric morphometric study to determine the feasibility of using such techniques in the general population. METHODS: A total of 709 atlas specimens were measured bilaterally to determine the thickness of the posterior-lateral arch at the level of the lateral mass. The presence of the ponticulus posticus was recorded as well. RESULTS: The average minimal thickness of the posterior-lateral atlas was 3.95 mm. There was a total of 157 complete and 34 incomplete ponticuli posticus. Overall, 85.2% of the specimens had a thickness greater than 3 mm, 654 (46.2%) greater than 4 mm, and 194 (13.7%) greater than 5 mm. CONCLUSIONS: Our results suggest that although only a small percentage of patients can accept a screw that is directly inserted via the posterior-lateral arch into the lateral mass, the notching technique is possible in the vast majority of patients. To our knowledge, this is the largest study to examine the possibility of using the posterior-lateral arch as the starting point for these screws. Our results suggest that alternative starting points for these screws are possible in a large percentage of the cases. PMID- 17108833 TI - Heterotopic ossification in total cervical artificial disc replacement. AB - STUDY DESIGN: Prospective clinical study enrolled in 2 centers (Munich and Liberec) as part of a prospective European multicenter study with ProDisc C (Synthes Inc., Paoli, PA). OBJECTIVES: The first goal of the study was to evaluate the rate of heterotopic ossifications identified with plain radiograph following total cervical disc replacement (TCDR). The second goal was to show whether segmental motion can be preserved, and whether TCDR can provide improvement of the patient's ability to perform activities of daily living as well as a decrease of pain. SUMMARY OF BACKGROUND DATA: Only a few reports about the radiologic outcome after TCDR are published so far. Heterotopic ossification is a well-known phenomenon after total hip arthroplasty. The rate of heterotopic ossification following TCDR is unclear. METHODS: The radiographs of 54 patients (in total, 77 implanted prostheses) were analyzed 1 year after TCDR with a ProDisc C prosthesis. We classified the heterotopic ossification in 5 grades according to a recently published classification system for lumbar total disc replacement. For clinical parameters, the visual analog scale and the Neck Disability Index were evaluated preoperatively and 1 year postoperatively. The Student t test and Wilcoxon test were used for statistical analysis. RESULTS: In 26 treated segments (33.8%), no heterotopic ossification was detectable. Grade 1 ossifications were present in 6 levels (7.8%). A total of 30 segments (39.0%) showed grade 2 ossifications. Heterotopic ossifications that led to restrictions of the range of motion were present in 8 cases (10.4%). One year postoperatively, 7 cases (9.1%) had a spontaneous fusion of the treated segment. The clinical parameters improved significantly and were similar to previous reports about TCDR. CONCLUSIONS: Only 33.8% of the patients did not show any signs of heterotopic ossification, and the rate of spontaneous fusion after TCDR 1 year after surgery was unexpectedly high. There were 49.4% of the patients with grade 2-3 ossification, which lets us suspect an even higher rate of spontaneous fusion after long-term follow-ups. Motion preservation after TCDR is only guaranteed if spontaneous fusion can be prevented. Thus, mobility of the implanted segments needs to be further studied. PMID- 17108834 TI - How does the ossification area of the posterior longitudinal ligament progress after cervical laminoplasty? AB - STUDY DESIGN: Retrospective case series. OBJECTIVE: To investigate the progression pattern of ossification of the posterior longitudinal ligament (OPLL) after cervical laminoplasty. SUMMARY OF BACKGROUND DATA: OPLL is a progressive disease, and an increased area of ossification affects the surgical results after laminoplasty. However, it is uncertain how the ossification area progresses with time after surgery. METHODS: Fifty-five patients who were available for serial radiographs of more than 5 years were included. The extent of ossification in the longitudinal axis was assessed using computer software. The associations between the progression of OPLL and the clinical and radiologic data were analyzed. RESULTS: Forty-one patients had OPLL progression. The patients were divided into three groups according to the pattern of progression. Group 1 consisted of patients 40 to 49 years of age with continuous or mixed type, showing slow progression at the beginning, then fast. Group 2 consisted of patients older than 50 years with continuous or mixed type, showing rapid progression at the beginning and then slow. The patients in Group 3 had segmental type with no or slight progression. CONCLUSIONS: We speculated that the progression of OPLL decreases as the patient ages. These findings will be important for the management of patients with OPLL after surgery. PMID- 17108835 TI - Increased swelling complications associated with off-label usage of rhBMP-2 in the anterior cervical spine. AB - STUDY DESIGN/SETTING: Independent, retrospective clinical record review with a concurrent control. OBJECTIVE: To identify whether rhBMP-2 is associated with an increased incidence of clinically relevant postoperative prevertebral swelling problems in patients undergoing anterior cervical fusions. SUMMARY OF BACKGROUND DATA: Bone Morphogenetic Protein-2 (rhBMP-2) is FDA approved as a bone graft substitute in anterior lumbar interbody fusions. rhBMP-2 has also been used "off label" in anterior cervical fusions. We suspected that rhBMP-2 might increase the incidence of adverse swelling events. METHODS: A total of 234 consecutive patients (ages 12-82 years) undergoing anterior cervical fusion with and without rhBMP-2 over a 2-year period at one institution comprised the study population. The incidence of clinically relevant prevertebral swelling was calculated. The populations were compared and statistical significance was determined. RESULTS: A total of 234 patients met the study criteria, 69 of whom underwent anterior cervical spine fusions using rhBMP-2; 27.5% of those patients in the rhBMP-2 group had a clinically significant swelling event versus only 3.6% of patients in the non-rhBMP-2 group. This difference was statistically significant (P < 0.0001) and remained so after controlling for other significant predictors of swelling. CONCLUSIONS: Off-label use of rhBMP-2 in the anterior cervical spine is associated with an increased rate of clinically relevant swelling events. PMID- 17108836 TI - Magnetic resonance imaging assessment of craniovertebral ligaments and membranes after whiplash trauma. AB - STUDY DESIGN: Review article. OBJECTIVES: To review the literature on soft tissue lesions of the upper cervical spine in whiplash trauma with focus on imaging. SUMMARY OF BACKGROUND DATA: Whiplash injury is associated with chronic impairment in a substantial number of patients. There are different opinions as to the nature and prognosis of this condition, and the role of diagnostic imaging is debated. METHODS: Review the literature on the anatomic source of the chronic whiplash syndrome. Review the literature on imaging of the upper cervical spine, emphasizing on the author's own research. RESULTS: MRI shows structural changes in ligaments and membranes after whiplash injury, and such lesions can be assessed with reasonable reliability. Lesions to specific structures can be linked with specific trauma mechanisms. There is a correlation between clinical impairment and morphologic findings. CONCLUSION: Whiplash trauma can damage soft tissue structures of the upper cervical spine, particularly the alar ligaments. Structural lesions in this area contribute to the understanding of the chronic whiplash syndrome. PMID- 17108837 TI - Use of computed tomography to predict failure of nonoperative treatment of unilateral facet fractures of the cervical spine. AB - STUDY DESIGN: Retrospective review. OBJECTIVES: To determine whether radiographic measurements derived from standard computed tomography (CT) evaluation can be used to predict failure of nonoperative treatment in patients with unilateral facet fractures. SUMMARY OF BACKGROUND DATA: There is no consensus regarding treatment of unilateral cervical spine facet fractures. Management of this injury is based primarily on the presence of neurologic deficits and the degree of perceived spinal instability. CT-based criteria for predicting failure of nonoperative treatment in this patient population have not been examined. METHODS: Initial CT scans of all patients with unilateral cervical facet fractures were reviewed. Direct measurements included height and width of the facet fracture fragment, fracture displacement, and angulation. Calculated data included percent height and width of the fracture fragment based on the height and width of the contralateral intact facet. RESULTS: A total of 24 patients with 26 unilateral facet fractures were identified. Five patients with 5 facet fractures failed nonoperative management and required delayed surgical stabilization. Comparing patients successfully treated nonoperatively to those failing nonoperative management, a significant difference was found in absolute height of the fracture fragment (P = 0.0002), articular fracture height (P = 0.008), and height of the fracture fragment expressed as a percentage of the contralateral intact lateral mass (P = 0.026). CONCLUSION: The only significant risk factors identified for failure of nonoperative treatment were craniocaudal height of the fracture fragment and relative height of the fracture fragment expressed as a percentage of the intact lateral mass. This study suggests that patients with unilateral cervical facet fractures involving >40% of the absolute height of the intact lateral mass or an absolute height >1 cm are at increased risk for failure of nonoperative treatment. Failure of nonoperative treatment was not observed in any patient with a fracture involving less than 40% of the height of the lateral mass or an absolute height <1 cm. PMID- 17108838 TI - The effect of lumbar flexion and extension on disc contour abnormality measured quantitatively on magnetic resonance imaging. AB - STUDY DESIGN: Experimental study with subjects as their own control. OBJECTIVE: To determine if lumbar disc contour abnormality dimensions, measured quantitatively, differ in flexed, neutral or extended positions. SUMMARY OF BACKGROUND DATA: MRIs obtained lying supine are used to determine the degree of lumbar disc contour abnormality (bulging or herniation). Variations in positioning are suspected to influence this assessment. METHODS: Lumbar MR images for 26 male volunteers (24-74 years of age), with or without low back pain, were obtained with the subjects lying in neutral, maximal flexion, and maximal extension positions allowable within a conventional 1.5T MR scanner. Quantitative measures of anterior and posterior disc contour abnormality were obtained for each position. RESULTS: Statistically significant differences in disc angles were obtained between positions (2-5 degrees) for all levels. Posterior contour abnormality was significantly smaller in flexion and extension than in the neutral position (9.5%-30.1%). Posterior contour abnormality in extension was similar or smaller than in flexion. Anterior contour abnormality was significantly smaller in extension than flexion and smaller in the neutral position than flexion. CONCLUSION: Spine position should be standardized when assessing disc contour abnormality with MRI. The largest measured disc contour abnormalities when lying supine in a standard MR scanner are observed in the neutral position, as opposed to flexion or extension. PMID- 17108839 TI - Prevalence and distribution of spinal osteoarthritis in women. AB - STUDY DESIGN: Retrospective review of lateral spinal thoracolumbar radiographs, obtained to rule out spinal injury after trauma, were scored for osteoarthritis. OBJECTIVES: The extent, prevalence, and distribution of spinal osteoarthritis in women aged 20-80 years was determined. SUMMARY OF BACKGROUND DATA: Radiographic evidence of disc space narrowing and osteophytosis is one method of assessing osteoarthritis, but population-based surveys of osteoarthritis have been limited due the dangers of exposing human subjects to radiographs. Consequently, the prevalence and the distribution of osteoarthritis in women have not been established. METHODS: Average individual disc space narrowing and osteophytosis scores were assessed using an atlas method. Prevalence was determined for decadal age groups and spinal level (T4-L7). RESULTS: The extent of osteoarthritis is weakly associated with age, while the prevalence of osteoarthritis increases with age. Two peaks in osteoarthritis prevalence are apparent: in the mid thoracic and lower lumbar spines. CONCLUSIONS: Although few younger women have high average scores, some older women have no radiographic sign of osteoarthritis, while others are severely affected. While the peak in osteoarthritis in the lumbar spine parallels the clinical symptom of low back pain, the more pronounced peak in the mid thoracic does not have a reported corresponding clinical symptom. PMID- 17108840 TI - Impact of surgical intervention on quality of life in patients with spinal metastases. AB - STUDY DESIGN: Prospective clinical study. OBJECTIVE: To assess Health-Related Quality of Life outcomes in patients undergoing surgery for spinal metastases. SUMMARY OF BACKGROUND DATA: Increasing life expectancy of patients with spinal metastases has resulted in greater interest in overall quality of life, including pain and neurologic impairment. To assess the overall risks and benefits of surgical intervention, the overall impact of each on the overall health status must be assessed. METHODS: All patients who presented to a single institution with bony spinal metastases requiring surgical intervention were eligible. EXCLUSION CRITERIA: previous surgery for spinal metastases, primary tumors of the spine, and inability to fill out the questionnaires. Patients completed an EORTC QLQ-C30, the HUI-3, the EQ-5D, visual analog pain, and an ECOG functional assessment. at five points: before surgery and at 6 weeks, 3 months, 6 months, and 1 year post surgery. RESULTS: Of 96 patients who presented to the hospital, 85 were enrolled in the study. Average age was 58.6 years (range, 20.3-80.7 years) with 47 male patients; 50% survival as 39.1 weeks. Maximal and average VAS pain levels showed a statistically significant (P < 0.00001) improvement from preoperative to all postoperative time points. Only the QLQ-C30 global health status showed a statistically significant improvement from preoperative to the 6 week (P = 0.017), 3-month (P = 0.039), and 6-month (P = 0.013) time points. There was a statistically significant correlation between baseline global health status and survival time (P = 0.041). Overall distribution of HUI-3 utility calculated Quality of Life Adjusted Years (QALY) during the 1-year postoperative period showed a bimodal distribution with peaks at 0.1 and 0.7 years. CONCLUSIONS: Surgery for patients with spinal metastases offers decreased pain and improved quality of life with low rates of surgical complications. PMID- 17108841 TI - Patient preferences regarding spine surgical decision making. AB - STUDY DESIGN: Patient surveys to determine preferences in surgical decision making. OBJECTIVE: To evaluate spine patient preferences regarding physician and patient roles in surgical decision making and to discuss the ethical considerations that arise. SUMMARY OF BACKGROUND DATA: Since the 1980s, there has been a push toward increasing patient autonomy and self-determination, and away from the paternalism of the past. Commensurate with this shift, patients have been encouraged to take the primary active role in surgical decision making. To date, there is little empirical evidence regarding how deeply patients want to be involved in this decision-making process. METHODS: A total of 200 consecutive patients seen at our academic spine center were administered 1 of 2 questionnaires (previously validated) aimed at determining patient preferences about how clinical decision making should take place. RESULTS: Patients felt strongly that complete risk information be provided. The majority of patients felt that the physician, rather than the patient, should make the basic treatment decision, and the great majority felt that the physician should make the technical decisions regarding treatment. CONCLUSIONS: Spine surgical patients often prefer to defer surgical decision making to their surgeons. In clinical scenarios where there is little controversy and the evidence is clear, this results in little consequence, assuming that the surgeon aims to provide evidence based care. In scenarios with greater controversy and less clear evidence, the choice of treatment offered by the surgeon may be based on factors outside of the available science, and, accordingly, efforts should be made to educate fully the patient and to help the patient make his/her own decision based on personal values regarding outcomes. PMID- 17108843 TI - Prediction of spinal canal expansion following cervical laminoplasty: a computer simulated comparison between single and double-door techniques. AB - STUDY DESIGN: Laminoplasty was simulated using a computer-assisted technique to assess the amount of canal expansion. OBJECTIVES: This study was designed to clarify the relationship between laminoplasty opening size and increase in sagittal canal diameter, increase in canal area, and the angle of the opened lamina following laminoplasty, and to determine whether a spinous process splitting laminoplasty achieves the similar canal expansion as a single open-door method. SUMMARY OF BACKGROUND DATA: Single and double-door cervical laminoplasty (SDCL and DDCL, respectively) have been widely used in the treatment of multilevel stenotic conditions. However, the relationship between laminoplasty opening size and spinal canal expansion following laminoplasty, and the comparison of postoperative spinal canal expansion between single and double-door techniques have not been well investigated. METHODS: SDCL and DDCL, based on preoperative computerized tomography scans of 34 patients who had undergone the laminoplasty surgery, were simulated using a computer-assisted technique. Laminoplasty with an opening size of 6, 8, 10, 12, 14, 16, and 18 mm were simulated to determine the amount of canal enlargement with the various opening size. RESULTS: Sagittal diameter, canal area, and lamina angle were increased steadily following either single or double-door laminoplasty with the door opened from 6 to 18 mm. Significant positive correlation was found between laminoplasty opening size and increase in sagittal diameter (R2 = 0.969 and P = 0.001 in SDCL; R2 = 0.926 and P < 0.001 in DDCL), increase in canal area (R2 = 0.961 and P < 0.001 in SDCL; R2 = 0.937 and P < 0.001 in DDCL), and lamina angle (R2 = 0.959 and P < 0.001 in SDCL; R2 = 0.943 and P < 0.001 in DDCL). No significant correlation was observed between preoperative sagittal diameter and increase in sagittal diameter of the spinal canal, whereas significant positive correlation was found between preoperative cross-section area and increase in cross-section area of the spinal canal. The differences between postoperative canal increase in sagittal diameter and canal area for the single versus double-door technique were statistically significant when the door was opened by more than 12 mm (P < 0.05). CONCLUSIONS: Our investigation provides insight into canal expansion after laminoplasty. The increased amount of canal following laminoplasty can be predicted by the regression equations. This may allow preoperative determination of the optimal size of the opening needed to establish adequate canal space for the spinal cord. Both single and double-door techniques of laminoplasty provide sufficient room for posterior migration of the spinal cord, although gaining different canal expansion. PMID- 17108844 TI - Re: Weinstein JN. An altruistic approach to clinical trials: the National Clinical Trials Consortium (NCTC). PMID- 17108845 TI - Sex hormone ratio changes in men and postmenopausal women with coronary artery disease. AB - OBJECTIVE: The goal of this study was to investigate the potential role of sex hormones in coronary atherosclerosis in both men and postmenopausal women. DESIGN: A total of 258 male and 236 female postmenopausal participants with angiographically defined stable coronary artery disease (CAD) were enrolled. We measured the levels of estradiol (E2), progesterone (P), testosterone (T), follicle-stimulating hormone, and luteinizing hormone in the participants and in 156 male and 132 female disease-free and age-matched controls using commercially available radioimmunoassay kits. RESULTS: In the male study participants and control subjects, the levels of E2 and P differed slightly in opposing directions; however, these differences were not significantly different, nor were there significant differences in T. However, the ratio of E2 to P in participants was significantly (P < 0.01) lower (even after adjustments for age and body mass index) than in the control subjects (mean +/- SEM: 70.2 +/- 56.4 vs 90.7 +/- 59.5, respectively). In the postmenopausal women, a slight decrease in E2 and increases in P and T in participants were not significantly different from levels in the control group. However, the E2 to P and E2 to T ratios were significantly (P < 0.01) lower (before and after adjustments for age and body mass index adjustments) in the participants relative to the control subjects (38.7 +/- 28.4 vs 49.6 +/- 36.3 and 46.5 +/- 37.6 vs 60.6 +/- 40.8, respectively). Correlation analyses demonstrated that the sex hormone ratio changes in both men and postmenopausal women were related with atherogenic blood lipoprotein changes. In both the male and female groups, levels of follicle-stimulating hormone and luteinizing hormone did not differ significantly between the participants and controls, and correlation analyses revealed no association between these hormones and the ratio of E2 to P in males and the ratios of E2 to P and E2 to T in females (r < 0.2, P > 0.05). Multiple regression analyses demonstrated that age and the presence of CAD were significantly and independently associated with the E2-to-P ratio in men and the E2-to-P and E2-to-T ratios in women and that E2-to-P ratio and low-density lipoprotein cholesterol level were significant independent predictors of CAD in males; E2-to-P and E2-to-T ratios and low-density lipoprotein cholesterol level were significant predictors of CAD in women. CONCLUSIONS: In both men and postmenopausal women with angiographic CAD, there were significant differences (relative to age-matched control subjects) in sex hormone ratios, suggesting an abnormality that could influence coronary health. A lower E2-to-P ratio may be associated with the male disposition to coronary atherosclerosis, whereas lower E2-to-P and E2-to-T ratios may be associated with the same condition in females. PMID- 17108846 TI - Comparison of patient recall of hormone therapy with physician records. AB - OBJECTIVE: The objective of this study was to compare patient recall of menopausal hormone therapy, obtained by personal interview, with that recorded in physicians' records. DESIGN: Data were from a case-control study of women aged 55 to 64 years at diagnosis of breast cancer in Los Angeles County, CA, between 1987 and 1989. Medical record information on 1,115 women reporting hormone therapy use (652 case patients and 463 control participants) was sought from 2,734 physicians and obtained from 1,350. Physician records for an additional 126 case patients and 90 control participants not reporting hormone therapy but reporting oral contraceptive use or benign breast disease were also obtained. RESULTS: Agreement between interview and physician records was 85% (kappa=0.59) for ever/never use of estrogen and 84% (kappa=0.59) for progestogen. The correlations for months of use were 0.43 and 0.34, respectively. For both drugs participants tended to report longer use, primarily because they reported earlier starting ages. Older and unmarried women recalled more unconfirmed estrogen use, whereas naturally menopausal women were more likely to report no estrogen use when the medical record was positive. We found no evidence of preferential recall bias by case patients. CONCLUSIONS: Interviews provide a moderately reliable measure for ever use of postmenopausal estrogen and progestogen. Recalled details are less reliable and may be affected by age, type of menopause, and recall interval. PMID- 17108847 TI - Testosterone inhibits estrogen/progestogen-induced breast cell proliferation in postmenopausal women. AB - OBJECTIVE: During the past few years serious concern has been raised about the safety of combined estrogen/progestogen hormone therapy, in particular about its effects on the breast. Several observations suggest that androgens may counteract the proliferative effects of estrogen and progestogen in the mammary gland. Thus, we aimed to study the effects of testosterone addition on breast cell proliferation during postmenopausal estrogen/progestogen therapy. DESIGN: We conducted a 6-month prospective, randomized, double-blind, placebo-controlled study. A total of 99 postmenopausal women were given continuous combined estradiol 2 mg/norethisterone acetate 1 mg and were equally randomly assigned to receive additional treatment with either a testosterone patch releasing 300 microg/24 hours or a placebo patch. Breast cells were collected by fine needle aspiration biopsy at baseline and after 6 months, and the main outcome measure was the percentage of proliferating breast cells positively stained by the Ki 67/MIB-1 antibody. RESULTS: A total of 88 women, 47 receiving active treatment and 41 in the placebo group, completed the study. In the placebo group there was a more than fivefold increase (P<0.001) in total breast cell proliferation from baseline (median 1.1%) to 6 months (median 6.2%). During testosterone addition, no significant increase was recorded (1.6% vs 2.0%). The different effects of the two treatments were apparent in both epithelial and stromal cells. CONCLUSIONS: Addition of testosterone may counteract breast cell proliferation as induced by estrogen/progestogen therapy in postmenopausal women. PMID- 17108848 TI - Estrogen-progestin therapy in women after stem cell transplant: our experience and literature review. AB - Women undergoing stem cell transplantation (SCT) are mostly young and have more than 90% probability of ovarian failure, which is often permanent. A woman's age, use of radiotherapy and alkylating chemotherapy, and the allogeneic type of transplant are associated with a higher rate of premature ovarian failure and worse residual ovarian function. Premature ovarian failure has serious systemic and psychological effects that may need treatment and should be managed by practitioners trained to treat this particular population of women. Ultrasonographic evidence of ovarian follicles is often associated with a future resumption of cycles, but there are no serum markers to predict the return of ovarian function in these patients. In our center, the rate of ovarian function recovery was 7% after allogeneic SCT and 25% after autologous SCT (P<0.05). There are no guidelines on how to manage premature ovarian failure induced by myeloablative treatments followed by SCT. Because of the likelihood of the need for long-lasting estrogen plus progestin therapy (EPT) and the increased risk of secondary neoplasia after SCT, the EPT should be as physiological as possible. In our experience, the cyclical sequential combination of estradiol (2 mg daily) plus dydrogesterone (10 mg for 14 d/mo) was associated with excellent compliance because of its simple administration and few adverse effects. Such a treatment led to a dramatic improvement in vasomotor, urogenital, and psychological symptoms related to estrogen deficiency. However, in the allogeneic transplantation setting, up to 25% of women may suffer from gynecological chronic graft-versus-host disease, which may become apparent as hematocolpometra after introduction of EPT. Thus, accurate pretreatment evaluation and frequent monitoring during treatment are required. Moreover, EPT absorption may be reduced in patients who received allotransplants and have gastrointestinal or skin chronic graft-versus-host disease. PMID- 17108849 TI - Risk factors for genital and anal warts in a prospective cohort of HIV-negative homosexual men: the HIM study. AB - OBJECTIVE: The objective of this study was to determine the prevalence, incidence, and risk factors for genital and anal warts in HIV-negative homosexual men in Sydney. STUDY DESIGN: The authors conducted a prospective cohort study. Participants were asked whether they had had genital and anal warts at each interview. Details of lifetime sexual contacts and sexual behaviors in the last 6 months were collected. RESULTS: Among 1,427 men recruited, 8.9% and 19.6% reported a history of genital and anal warts at baseline, respectively. Incidence rates for genital and anal warts were 0.94 and 1.92 per 100 person-years, respectively. In multivariate analysis, both incident genital and anal warts were associated with younger age. In addition, incident genital warts was associated with insertive fingering (P trend = 0.018), whereas incident anal warts was associated with insertive fingering (P trend = 0.007) and insertive fisting (P trend = 0.039). CONCLUSIONS: Anal warts were twice as common as genital warts. Fingering and other manual sexual practices may be an important transmission route for both. PMID- 17108850 TI - Reliability and criterion-related validity of self-report of syphilis. AB - OBJECTIVE: To determine the test-retest reliability, sensitivity and specificity, and criterion-related validity of the Risk Behavior Assessment (RBA) syphilis questions. The RBA is a standardized instrument that has been used in several studies of STDs in drug users. METHODS: For the test-retest reliability study, 219 injection drug users completed the RBA twice within a 48-hour period. To determine criterion-related validity, 207 individuals, who also completed the RBA, were tested with the rapid plasma reagin test (RPR), and 206 individuals were also tested with the Serodia Treponema pallidum particle agglutination test (TP-PA). RESULTS: The test-retest reliability for the question "How many times have you been told by a doctor or a nurse that you had syphilis?" was 0.78. The test-retest reliability for the question "In what year were you last treated for syphilis?" was 0.89. For the comparison of self-report with the RPR test, the sensitivity of self-report was 46.2% and the specificity was 95.7%. For the comparison of self-report with the TP-PA test, the sensitivity of self-report was 37% and the specificity was 97.7%. CONCLUSIONS: Self-reports of syphilis infection history were found to have good reliability, excellent specificity, and moderate sensitivity. These characteristics need to be taken into account in any study using these self-report items. PMID- 17108851 TI - A continuing dilemma for recently qualified nurses. PMID- 17108852 TI - HCAIs can be minimized through effective teamwork. PMID- 17108853 TI - Creating the evidence base: the journey from practice to research. AB - The ability to deliver accurate, evidence-based information when helping patients make decisions about their care is fundamental to the role of the clinical nurse specialist. For patients with ulcerative colitis who are considering surgery to form an ileo-anal pouch, it is essential to give them information about the psychosocial, as well as the physical, effects of such surgery. As many of these patients are of working age, understanding the likely consequences for paid employment is a priority. However, very little evidence is available on this subject. The authors therefore decided to develop a research study aimed at filling this gap. A proposal to conduct a phenomenological study into the subsequent employment experiences of patients who have undergone ileo-anal pouch formation has been developed. This article seeks to explore how the research idea and its methodology stemmed from practice and highlights the importance of seeing research and practice as complementary components of good nursing care. PMID- 17108854 TI - Alternative management of complex wounds and fistulae. AB - The management of complex wounds and fistulae can often prove challenging to even the most skilled clinician. The incidence and complexity of fistulae vary considerably from centre to centre, however they often lead to prolonged hospital stays. Routine admissions for 4-5 days may lead to 4-5 months in the event of fistulae formation. Thus, many patients experience not only compromised physical health, but also complex psychological problems. This article provides a brief overview of the challenges and developments of managing a complex wound with multiple fistulae and a pictorial illustration of an innovative alternative wound management system. PMID- 17108855 TI - Study of peristomal skin disorders in patients with permanent stomas. AB - The aim of this article was to investigate the frequency, severity and diversity of peristomal skin disorders among individuals with a permanent stoma in a community population. All individuals with a permanent stoma (n=630) in a Danish community population were invited to participate in a cross-sectional study. A total of 202 individuals (101 men; 101 women) agreed to participate. Data were collected through questionnaires and clinical examinations. It was found that peristomal skin disorders were higher for participants with an ileostomy (57%) and urostomy (48%) than in those with a colostomy (35%). Of the diagnoses of skin disorders, 77% could be related to contact with stoma effluent. Only 38% of diagnosed participants agreed that they had a skin disorder and more than 80% did not seek professional health care. The study revealed a high frequency of peristomal skin disorders. Participants frequently failed to perceive that they had a skin irritation and did not seek help. This suggests that more education and perhaps regular, annual follow-up visits at local stoma care clinics are needed. PMID- 17108856 TI - Professional expectations outside the work setting. PMID- 17108857 TI - Review and discussion of the body's normal microorganisms. AB - Nurses need to be constantly vigilant in their hygiene practices, yet not ignore the role of normal microbes in the maintenance of health: we have more microorganisms living on us and in us than any other type of cell, and they have a role in preventing colonisation by harmful pathogens, and also in regulating the immune system. This article reviews the types of microorganisms normally found in and on individuals, and how these can contribute to health. PMID- 17108858 TI - Safety in urine sampling: maintaining an infection-free environment. AB - The insertion of an indwelling urinary catheter is thought to be implicated in 80% of all urinary tract infections (UTI). In patients with an indwelling catheter, one of the most effective ways to determine whether or not there is evidence of a UTI is to aspirate a sample of urine for analysis, typically using a syringe and needle. Inevitably this puts the healthcare worker at risk of a percutaneous needle stick injury and accidental exposure to serious and potentially life-threatening viruses. Infection control is now taken increasingly seriously in UK hospitals, and the elimination of infection risk is regarded as a priority. In addition to training, and safer working practices the government recognizes the need for new technological innovation to help further reduce risks. In the urine sampling environment, there is a new needle-free port (EZ-Lok from Bard Ltd), designed specifically to reduce the occupational health risk of needle stick injuries. PMID- 17108859 TI - Chronic obstructive pulmonary disease: clinical signs and infections. AB - Chronic obstructive pulmonary disease (COPD) is a pathological condition that involves excessive production of mucus, chronic cough and inflammatory changes leading to airway limitation. The most common cause of COPD is cigarette smoking. COPD can be categorized into either chronic bronchitis or emphysema; both conditions can be differentiated by age and the production of copious, tenacious sputum. Patients with COPD are susceptible to pulmonary infections of bacterial or viral origin. Nurses should be aware of the subtle differences in conditions and the usefulness of antibiotics in the treatment of acute bacterial exacerbations of chronic bronchitis. PMID- 17108860 TI - Medicines for hypertension: changes to the UK NICE guidelines. AB - New NICE guidance on the use of medicines for hypertension, published in June 2006, will prompt a major review of treatment for those already receiving medication and also sets out a new treatment pathway for the newly diagnosed. The major change in the guidelines is in the status of beta blockers, which are no longer to be used as a routine treatment for hypertension. The guidelines provide a consensus on treatment, and nurses and their patients now have a clear and straightforward algorithm to follow. It is envisaged that these changes to the way medicines are prescribed for hypertension will have a significant impact on the number of people reaching their target blood pressure, and therefore the number of cardiovascular events occurring each year. PMID- 17108861 TI - Developing a plan of care using the Roper, Logan and Tierney model. AB - This care study, using the Roper, Logan and Tierney Model of Nursing, examines the presentation of a patient with a history of drug misuse and a current diagnosis of infective endocarditis and hepatitis C. While this particular model of nursing incorporates 12 activities of daily living, this care study will deal with three specific activities: maintaining a safe environment, communication and breathing. One of the main concerns arising from this care study was the inadequate management of pain for the patient. It is suggested in the literature that healthcare professionals do not always adequately address pain management, primarily because of fear of causing addiction. The authors suggest that the patient's history of drug misuse in this study may have further mitigated against him receiving adequate pain control. PMID- 17108862 TI - Consent to use complementary therapies on critical care wards. AB - This article explores the use of complementary therapies as part of care within the NHS and the legal issues which arise. The employer is entitled to give or refuse consent to the use of complementary therapies by its employees on NHS patients and to lay down the conditions on which such therapies could be practised. PMID- 17108863 TI - How prepared are nurses for terrorist attacks? PMID- 17108864 TI - Effect of a symbiotic preparation on the clinical manifestations of irritable bowel syndrome, constipation-variant. Results of an open, uncontrolled multicenter study. AB - AIM: Irritable bowel syndrome (IBS) is frequently associated with an imbalance in intestinal bacteria. To date, few studies have evaluated the efficacy and safety of probiotic administration in patients with constipation-variant IBS. A new agent recently available in clinical practice is a symbiotic consisting of a probiotic, Bifidobacterium longum W11, and the short chain oligosaccharide prebiotic Fos Actilight. The aim of this study was to evaluate the efficacy and safety of this symbiotic in patients with constipation-variant IBS. METHODS: A total of 636 patients (250 men, 386 women) diagnosed with constipation-type IBS according to the Roma II criteria were enrolled in 43 centers and received the symbiotic at a dose of 3 g/die for at least 36 days. A validated questionnaire investigating symptoms and stool frequency was administered before and after treatment. RESULTS: Based on patient responses to visual scale items, frequency increased significantly after treatment in the ''no symptom'' class from 3% to 26.7% for bloating and from 8.4% to 44.1% for abdominal pain (P<0.0001). In the more severe symptoms classes (moderate-severe), symptom frequency dropped significantly from 62.9% to 9.6% and from 38.8% to 4.1% for bloating and abdominal pain, respectively. Stool frequency significantly increased from 2.9+/ 1.6 times/week to 4.1+/-1.6 times/ week. CONCLUSIONS: The study product can increase stool frequency in patients with constipation-variant IBS and reduce abdominal pain and bloating in those with moderate-severe symptoms. PMID- 17108865 TI - Treatment of irritable bowel syndrome. A case control experience. AB - AIM: As optimal therapy for irritable bowel syndrome (IBS) remains elusive, current approach to therapy is based on symptomatic treatment. With this case control experience we wanted to determine the beneficial effect in IBS patients of a dietary integrator (IBS Active), composed of L-tryptophan, inulin, angelica, vegetal charcoal, vitamin PP, group B vitamins (B1, B2, B6) and probiotics (Lactobacillus sporogenes, Lactobacillus acidophilus, Streptococcus thermophilus). METHODS: The treatment group comprised 37 patients (11 men and 27 women; mean age, 44.3+/-5.1 years) given IBS Active (440 mg bid) over a mean period of 6 months (range, 5-8). The control group comprised 28 patients (6 men and 22 women; mean age, 48.6+/-3.7 years) who were instructed to continue their customary therapy for 6 months (range, 5-7). All subjects were assessed for the presence of abdominal pain and/or distension, constipation, diarrhea and alternating constipation and diarrhea. RESULTS: Compared with baseline values, the reduction in abdominal pain in the treatment group was 62% (P<0.0001), 55% (P<0.0001) in abdominal distension, 58% (P=0.05) in constipation, 33% (P=0.3) in diarrhea, and 62% (P=0.01) in alternation constipation and diarrhea. Compared with baseline values, no statistically significant reduction in symptoms was found in the control group. Post-treatment comparison between the two groups showed that the study product had reduced symptoms and that the difference was statistically significant for abdominal pain (P<0.000001), abdominal distension (P=0.003) and constipation (P=0.03). CONCLUSIONS: The use of IBS Active led to a significant improvement in pain symptoms, abdominal distension and regulation of bowel movement in IBS patients. Further study is needed to evaluate the long-term benefit of the study product. PMID- 17108866 TI - Biophysics and clinical practice for regenerative processes in cirrhosis of the liver/of liver cirrhosis assisted by Delta-S Entropy Variation Systems. AB - AIM: The therapy in question uses an innovative bioengineering device denoted as ''Delta-S DVD Entropy Variation System''. Previous research indicated regression of cirrhosis as evaluated in its morphofunctional and symptomatological aspects. The aim of the study is to confirm and extend previous experimental observations by enhancing hemodynamic evaluation techniques. In order to clarify scar regression, it was decided to include in the endpoints a quantitative evaluation of portal hypertension called HVPG, which is sensitive to the breakdown of hepatic architecture and the influence of regeneration nodules and therefore the advance of cirrhosis. METHODS: The experimental design consists of a self controlled study carried out on Child A-B cirrhosis patients with portal hypertension (hepatic venous pressure gradient, HVPG > or = 10 mmHg). Five patients were enrolled, 4 HCV positive, one with autoimmune cirrhosis, all showing extensive symptoms. RESULTS: At the end of the treatment all patients showed a reduction in portal hypertension (mean reduction HVPG = 40.2%, P<0.011), together with an improved ultrasound flowmeter pattern and a sharp decrease or disappearance of the symptoms. No adverse effects were reported. Efficacy on autoimmune cirrhosis was unaffected. CONCLUSIONS: By means of a quantitative analysis of portal hypertension and of functional aspects, this study confirms that the Delta-S DVD system can lead to the regression of the scar component of cirrhosis, promote the regeneration of functioning liver tissue with positive effects on hepatic functionality and prevent symptoms and the risk of varicose vein rupture. PMID- 17108867 TI - Evaluation of paraumbilical vein as a prognostic index of severe liver cirrhotic patients with portal-systemic shunts. AB - AIM: The aim of this study was to predict the outcome in severe liver cirrhotic patients with portal-systemic shunts. METHODS: One-hundred and sixteen patients with liver cirrhosis diagnosed as Child-Pugh class B and C with portal-systemic shunts confirmed by abdominal ultrasonography, computed tomography and magnetic resonance imaging were enrolled in this study. Twenty-three factors were evaluated concerning clinical laboratory parameters and extracted prognostic factors using the Cox proportional hazards model, and the prognostic index (PI) was prepared by combining these factors. RESULTS: The cumulative survival rates after admission were 64.6%, 35.6% and 25% after 1, 3 and 5 years, respectively. Using multivariate analysis, age, the presence of hepatocellular carcinoma (HCC), portal vein tumor thrombosis (PVTT) and paraumbilical vein (PUV) shunt were selected as significant prognostic factors that contributed independently to the prognosis of severe liver cirrhotic patients with portal-systemic shunts. The PI was calculated with the following formula using these 4 factors. PI = 0.042 x Age + 0.913 x HCC + 0.989 x PVTT + 1.079 x PUV shunt. The group with a high score for PI was found to die with significantly higher frequency than the group with a low score. CONCLUSIONS: It was found that tumor related factors and PUV shunt were the most important factors for severe liver cirrhotic patients with portal systemic shunts. The PI is suggested to be an appropriate index to predict the prognosis for these patients. PMID- 17108868 TI - Obesity and related diseases: an epidemiologic study in eastern Sicily. AB - AIM: The aim of this study was to investigate the prevalence of obesity in a non selected cohort of adult subjects living in eastern Sicily. METHODS: Out of 2 296 examined subjects, 834 (36.3%) were affected by obesity. Of these, only 160 (19.1%) were affected by obesity alone while 674 (80.9%) showed other associated pathologies. RESULTS: The prevalence of arterial hypertension, diabetes mellitus, hepatic steatosis, hyperdyslipidemia and renal failure was significantly higher (P = 0.000) than in a control group of non-obese subjects comparable for sex and age. CONCLUSIONS: In a large part of obese patients, the presence of insulin resistance was observed suggesting that this alteration can play a pivotal role in the development of some important metabolic and cardiovascular complications related to obesity. PMID- 17108869 TI - Molecular targets for therapy of hepatitis B virus-induced hepatocellular carcinoma. AB - Chronic hepatitis B virus (HBV) infection is the most frequent risk factor for development of hepatocellular carcinoma (HCC) worldwide. Studies of the molecular genetics and pathophysiology of HCC suggest that there are significant differences in the allelic imbalance, genome copy number, and gene expression patterns of HBV-induced HCC as compared to HCCs from other causes, which are presumably reflected to differences in the mode of presentation and outcomes of HBV-induced HCCs. Unique features of HBV-induced carcinogenesis include the role of HBV DNA integration in carcinogenesis and the powerful synergism between HBV and dietary aflatoxins in the pathogenesis of HCC. A more complete understanding of the biology of HBV-induced HCCs may reveal well-defined differences in the molecular pathways that regulate growth of these HCCs and allow better-targeted approaches to prevention and therapy of HBV-induced HCCs. This review will attempt to summarize the current knowledge about carcinogenic pathways in HBV induced HCCs, review the agents currently in development for targeted therapy of HCCs, and propose potentially novel approaches to therapy of HBV-induced HCC. PMID- 17108870 TI - Cell-matrix talks: effects on differentiation of hepatocellular carcinoma. AB - Different event is a process that is dependent on stimulation of extracellular signals, signal transduction and gene express. Malignant transformation of hepatocytes may occur in cirrhosis, which is the result of extracellular matrix (ECM) remodeling. ECM could affect and maintain the differentiated phenotype of hepatocytes by regulating liver transcription factors. Moreover, ECM remodeling is correlated with dedifferentiation of hepatocellular carcinoma (HCC). Integrin matrix adhesion system and E-cadherin/catenin adhesion complex mediate the cell matrix interaction through focal adhesion kinase, extracellular-signal-regulated kinases and beta catenin/Wnt pathway. The different event of HCC compared with the reversion of abnormal cell-matrix interaction. New drugs that are power for regulating cell-matrix interaction may represent a novel therapeutic strategy for HCC. PMID- 17108871 TI - Medical strategies for weight loss in the overweight and obese patient. AB - In recent years, obesity has become a major public health problem in Western countries. The World Health Organization has defined obesity as a global epidemic of the third millennium. Treatment options for weight management include dietary intervention, physical activity, behavior modification, pharmacotherapy and surgery. However, the complexity of this chronic condition necessitates a coordinated multidisciplinary team-approach to the care of obese patients who fail weight control. The long-term duration of the treatment and the necessity of monitoring compliance and effectiveness should be considered. The objective of this article was to review the major controlled randomized clinical trials dealing with the different medical strategies for weight loss and its maintenance in overweight and obese patients. PMID- 17108872 TI - Chronic anorexia nervosa: enteral nutrition via percutaneous endoscopic gastrostomy and liaison psychiatry. AB - Anorexia nervosa is a complex mental disorder characterized by altered eating behaviour often resulting in life-threatening weight loss (<85% of expected body weight) associated with amenorrhea and a disturbance of body image. Although classified as mental health disorders, they may lead to serious medical consequences and have the highest rate of premature death of any mental health diagnosis. We report our experience with the use of enteral feeding via percutaneous endoscopic gastrostomy in a 39-year-old woman with chronic restricter anorexia nervosa treated in liaison psychiatry and psychotherapy. On admission to psychiatry unit, the patient presented seriously deteriorated general condition and a body mass index (BMI) of 10 (BMI = weight kg/height m(2)). She refused oral feeding, but eventually accepted nasogastric feeding. In preparation for her continuing long-term (>1 month) enteral feeding at home, a percutaneous endoscopic gastrostomy was performed and a home nutrition support regimen that met her energy-protein intake requirements was prescribed. During the follow-up period, an overall improvement in nutritional status, general condition, mood and cognitive functioning was observed. Patient compliance with refeeding is notoriously problematic; however, enteral feeding interventions may be feasible in the long-term treatment of selected anorexia nervosa patients when closely followed-up by a multidisciplinary medical team. PMID- 17108873 TI - Absence of gene mutations in a case of concomitant presence of carcinoid of the ampulla of vater pheochromocytoma and Von Recklinghausen disease. AB - The rare association between Von Recklin-ghausen's disease (VRD) and tumours other than in central nervous system is well recognized. However, the concomitance of VRD, a carcinoid of the ampulla of Vater, and a pheochromocytoma has been described very rarely in literature. Furthermore, the possible role of the genes usually involved in multiple endocrine neoplasia (MEN) syndromes, in this association, is unclear. We report the case of a patient affected by VRD and extra-adrenal pheochromocytoma, operated on in the past for a carcinoid of the ampulla of Vater. To determine if genes involved in MEN syndromes might play a role in this particular triad, we investigated the presence of somatic or germline mutations in the RET proto-oncogene and menin gene by non isotopic polymerase chain reaction single stranded conformation polymorphism (PCR-SSCP) and heteroduplex gel electro-phoresis. The results demonstrated that no somatic or germline mutations in the MEN-1 and MEN-2 genes were involved in the pathogenesis of these tumours. PMID- 17108874 TI - Crohn's disease and Takayasu's arteritis: is this association difficult to find out? AB - Takayasu's arteritis is rarely reported associated with Crohn's disease in the English world literature. A case of Takayasu's arteritis type V in a young female affected by Crohn's disease is presented and the pathophysiology mechanism, and the clinical and diagnostic aspects are underlined. Surgical strategy is discussed. PMID- 17108875 TI - [Infertility: a global perspective]. AB - Infertility has always existed; however, today it has a different meaning because the possibility exists, not only to treat it in some three quarters of all cases, but also, in a number of instances, to prevent it. At the same time, this improved scenario created a number of important new issues concerning public health policies and the social impact of infertility; these issues can be summarized with two words: equity and ethics. Indeed, there are intolerable differences in access to infertility care, depending on the social-economic status, as well as the country in which a couple lives. It is generally believed that, overall, in the general population a persistent form of infertility affects some 7-8% of all couples; in Europe the prevalence of infertility has been estimated at around 14%. There are important regional differences in the incidence and causes of sterility. Whereas the best known factor in western countries is an increase in the age in which women attempt to conceive, in Africa, the most important cause is represented by sexually transmitted infections. Over the last fifty years there have been major break-throws in the diagnosis and treatment of infertility, although it must be stressed that the importance and validity of therapy must be in all cases evaluated against expectant management. This is because in couples without clear signs of functional or organic pathology, conception will occur spontaneously in between 80 and 90% of the cases. At the same time, if attempts to conceive fail over a period of three years, even in women 30 years or younger, the probability of pregnancy decreases to about 40%. Finally, it must be stressed that advances in assisted reproductive technology have created not only important successes, but also significant ethical problems: on the one hand, the debate is open on the limits of artificial interventions in the field of reproduction; on the other the possibility exists that a partner may ''impose'' an infertility on the other. PMID- 17108876 TI - [Up-date on cytoreductive surgery in the management of advanced ovarian cancer]. AB - Epithelial ovarian cancer represents the most aggressive neoplasm of women genital apparatus with a total 5-year survival rate ranging from 17% to 35% if the disease is in the metastatic phase. Its aggressiveness derives from the fact that it is an asymptomatic disease until it spreads in abdominal cavity. Therefore, in 70% of the cases, the diagnosis is done when tumor is already in advanced phase (Stage FIGO IIB-IV). Data from international literature suggest that standard treatment for advanced ovarian cancer is optimal cytoreductive surgery with adjuvant chemotherapy platinum-based. However, in the last decades, many authors have described the enthusiastic results of neoadjuvant chemotherapy and interval debulking surgery. Griffiths, first, underlined the importance of residual mass after cytoreductive surgery as a prognostic factor. Currently, cytoreduction is defined optimal when residual mass is microscopical or absent. Nevertheless, surgery for ovarian cancer turns out to be a particularly aggressive surgery that needs an operator's remarkable technical ability and a cultural Background: Many studies demonstrated that the frequency of feasibility of optimal cytoreductive surgery also varies within the gynecologic oncology specialized centers. During the last few years, new technologies (such as Cavitron Ultrasonic Surgical Aspirator, CUSA, and argon's coagulator) and new surgical techniques have been introduced. Ovarian cancer turns out to be a particularly chemosensitive tumor. Its responsiveness has been the object of numerous studies and protocols in literature, such as European Organisation of Research and Treatment of Cancer (EORTC) and Gynecologic Oncology Group (GOG) trials. PMID- 17108877 TI - [Immunological abortion: the thyroid factor]. AB - Spontaneous miscarriage (SM) is a multifactorial problem involving several couples. Recent studies investigated the correlations between the presence of antithyroid antibodies (ATA) and pregnancy loss, and found that many women with a previous history of recurrent miscarriage, showed high levels of ATA circulating in their blood. Further-more, the thyroid function disorder may also affect the course of pregnancy. Basically, two theories can explain the reasons of the spontaneous termination of pregnancy in presence of ATA: the first theory suggests that the hypofertlity or infertlity of these subjects may be due to a subtle degree of hypothyroidism which is difficult to detect by routine serum hormone determinations; the second theory supports that the presence of thyroid antibodies reveals a more generalized underlying abnormal stimulation of the immune system. Therefore, the thyroid function should be tested before conception and during pregnancy to avoid the pregnancy loss and neuropsychological deficits in infants. Actually, some papers suggest that treatments reserved to women with thyroid antibodies could decrease the miscarriage rate. Unfortunately, there is not agreement about the most effective therapy. We need more large, randomised, placebo controlled, double blind studies. PMID- 17108878 TI - [Tocolysis: which are the benefits?]. AB - Spontaneous preterm labor is still a major problem in perinatal medicine and it is associated to overwhelming risks of neonatal morbidity and mortality. Delaying delivery for hours, days or sometimes weeks may greatly reduce the short- and long-term perinatal morbidity improving fetal maturity of several organs and systems. Inhibition of uterine contractions may in fact allow to take advantage of the prenatal administration of glucocorticosteroids, which have been shown to reduce the incidence and severity of newborn respiratory distress syndrome. Moreover, delaying delivery may allow the transfer in utero to a maternity attached to a neonatal intensive care unit, therefore guaranteeing the best care for the preterm newborn. Every day gained between 22 and 28 weeks gestation increases survival by 3%. However, since most of preterm deliveries happened to be after 29 weeks, newborn survival is a secondary issue while the main aim of delaying labor in these cases is to improve the function of fetal systems and to try to understand if it is the case of prolonging pregnancy balancing risks of a hostile intrauterine environment towards the possible complications of a premature extrauterine life. PMID- 17108879 TI - [Ovulation induction in anovulatory women]. AB - Ovulation induction therapy is administered to stimulate follicular growth and induce ovulation in anovulatory infertile women. In anovulatory women with polycystic ovary syndrome, the treatment of choice is clomiphene citrate, whereas in clomiphene nonresponders, gonadotrophins are given as secondary therapy. Currently, insulin-sensitizing agents are used in the treatment of polycystic ovary syndrome to restore menstrual cyclicity. In selected patients, laparoscopic drilling has also been suggested. In anovulatory patients affected with hypogonadotropic hypogonadism, treatment is based on gonadotrophin replacement therapy or pulsatile gonadotrophin-releasing hormone infusion. In ovulation induction therapy the clinician's attention should be directed at restoring normal ovary function. When pharmacotherapy is required, monofollicular growth should be induced to reduce the risk of multiple pregnancy. PMID- 17108880 TI - [Progestogens and estroprogestins in the treatment of pelvic pain associated with endometriosis]. AB - We performed a MEDLINE and EMBASE search to identify all studies published in the English language literature on the use of progestogens for the treatment of endometriosis. The aim of our review was to clarify the biological rationale for treatment and define the drugs that can be used. It has been demonstrated that progestogens may prevent implantation and growth of regurgitated endometrium by inhibiting the expression of matrix metalloproteinases and angiogenesis, and they have several anti-inflammatory in vitro and in vivo effects that may reduce the inflammatory state generated by the metabolic activity of the ectopic endometrium. Oral contraceptives increase the abnormally low apoptotic activity of the endometrium of patients with endometriosis. Moreover, anovulation, decidualization, amenorrhoea and the establishment of a steady estrogen progestogen milieu contribute to disease quiescence. Progestogens are able to control pain symptoms in approximately three out of four women with endometriosi. Different compounds can be administered by the oral, intramuscular, subcutaneous, intravaginal or intrauterine route, each with specific advantages or disadvantages. Medical treatment plays a role in the therapeutic strategy only if administered over a prolonged period of time. Given their good tolerability, minor metabolic effects and low cost, progestogens must therefore be considered drugs of choice and are currently the only safe and economic alternative to surgery. However, their contraceptive effectiveness limits their use to women who do not wish to have children in the short-term. PMID- 17108881 TI - Endometrial stem/progenitor cells and proliferative disorders of the endometrium. AB - The human endometrium undergoes cyclical processes of regeneration, differentiation and shedding as part of the menstrual cycle. In non-menstruating species, there are cycles of endometrial growth and apoptosis rather than physical shedding. It was hypothesized many years ago that endometrial stem cells are responsible for the remarkable regenerative capacity of endometrium. In this review, we summarize the first data providing evidence for the presence of adult stem/progenitor cells in human and mouse endometrium using functional assays. This is because adult stem cells are defined by their functional properties rather than by marker expression, and there are no known markers of endometrial stem/progenitor cells. Evidence will be presented which demonstrates that the endometrium contains rare populations of both and epithelial and stromal stem/progenitor cells. These fundamental studies on endometrial stem/progenitor cells will provide new insights into the pathophysiology of various gynecological disorders associated with abnormal endometrial proliferation, including endometriosis, endometrial cancer, endometrial hyperplasia, and adenomyosis. The possible use of endometrial stem/progenitor cells in tissue engineering applications relevant to urogynecology will also be mentioned. PMID- 17108882 TI - Serum and peritoneal abnormalities in endometriosis: potential use as diagnostic markers. AB - Endometriosis is an ambiguous disease and its exact pathogenesis still remains elusive to clinicians and scientists. Local and systemic aberrations in immune response are associated with endometriosis. This article reviews the literature regarding various immunological factors such as cytokines, growth factors, adhesion molecules and angiogenic factors involved in the etiopathogenesis of this disease. Our review summarizes the literature regarding biomarkers, which may be reliable nonsurgical tools used in the diagnosis of endometriosis. Superior biomarkers characterized by high sensitivity, specificity and predictive value can help in the early detection and monitoring of disease progression as well as its response to therapeutic treatments critical for its management. A combination predictive model utilizing multiple biomarkers rather then individual markers alone is proposed to improve the diagnostic performance for identifying women with a high likelihood of having endometriosis. Immunomodulators and angiogenic factor blockers have a potential for endometriosis treatment and also to alleviate the pain or infertility associated with the disease. Potential new therapeutic agents include modulators, such as cytokine receptor blockers and angiogenic receptor blockers, presently used for treating endometriosis. PMID- 17108883 TI - [GnRH agonists and antagonists in the preoperative therapy of uterine fibroids: literature review]. AB - Uterine leiomyomas are the most frequent gynecological benign tumors. Their growth is regulated by ovarian steroids, therefore a hypoestrogenic state, like menopause or pharmacologically induced pseudo-menopause by GnRH-agonists or GnRH antagonists, is associated with the decrease of their volume. This volume reduction allows a less invasive surgical procedure and may reduce the amount of blood loss during surgery. Therefore, GnRH-agonists and antagonists are used in presurgical treatment of uterine fibromatosis. This review analyses the effects of GnRH-agonists and GnRH-antagonists therapies. GnRH-agonists produce a down regulation of GnRH receptor, while GnRH-antagonists compete with endogenous GnRH for pituitary binding sites. Due to the lack of any intrinsic activity of GnRH antagonists, the characteristic initial flare-up observed with GnRH-agonist treatment is absent. So, GnRH-antagonists rapidly suppress gonadotropin release within 4-8 h, while GnRH-agonists show clinical effects after 2 or 3 weeks of treatment. GnRH-antagonist activity is dose-dependent so it is possible to adjust the dose to obtain the proper levels of inhibition. The GnRH-agonist presurgical treatment usually is a short-term therapy (3-6 months), because it causes side effects like menopause symptoms. GnRH-antagonist clinical effects can be achieved with a short-time therapy too. Their side-effects include flushes and head-ache. After stopping therapy with both drugs, leiomyomas rapidly achieve their original size while side-effects disappear. Further studies are necessary to establish the use of GnRH-antagonists in leiomyomas therapy, but in Italy this is not possible because their use is not approved. PMID- 17108884 TI - Assessment of metabolic cerebral damage using proton magnetic resonance spectroscopy in mild traumatic brain injury. AB - AIM: We want to appraise, through proton magnetic resonance spectroscopy (H1 MRI), the NAA's values and it's changing in cerebral tissue in consequence of cranial trauma. METHODS: Six patients with TBI undergo to H1-MRI to asses the changes occurring briefly after trauma in the spectrum's composition. RESULTS: As far as the first two cases we founded a lowering of the NAA's values. In the other four cases the NAA values were normal in all but one, which slightly brought the values of the NAA/Cr and NAA/Cho lower in comparison to the standard values. CONCLUSIONS: Existing a correlation between NAA and ATP it can be drawn that the reduction of NAA is correlated to energetic type damage. Despite the smallness of data, it remains really important that we should have a tool to monitor the cerebral metabolic picture after a mild trauma. PMID- 17108885 TI - A new animal model for monitoring the early cerebral vasospasm after subarachnoid haemorrhage. AB - AIM: Spasm of cerebral arteries is a complication associated with subarachnoid haemorrhage. The aim of the present work is to find an experimental model of reliable, simple and in vivo monitoring of ''early'' basilar artery spasm after subarachnoid haemorrhage (SAH). Early spasm occurs within minutes of the SAH, its duration is approximately 1 hour. The need of different morphological and haemodynamic methods to evaluate experimental early spasm is reported. METHODS: To overcome intracranial surgical manipulations and biological of contrast and fixation media we designed a model that allows in vivo functional monitoring of basilar blood flow far away from the spasm without direct surgical and chemical interferences. Seventeen adult Burgundy rabbits were studied. RESULTS: Under homeostatic monitoring ''on-line'' carotid blood flow (carotid BF) .changes produced by SAH in cisterna magna of 12 (plus 5 sham treated) animals were studied from the common carotid artery after external carotid artery occlusion before, during SAH up to the end of the experiments. All the animals underwent digital subtraction cerebral panangiography (CPA) after SAH obtaining a significant increase of carotid BF only when basilar vasospasm was shown by CPA. CONCLUSIONS: Carotid BF increase during basilar vasospasm was defined ''functional. monitoring'' of early spasm. PMID- 17108886 TI - Unruptured Aneurysms Italian Study (UAIS) - background and method. AB - Treatment of unruptured cerebral aneurysms still represents an unsettled question in neurosurgical and neuroradiological communities. Although nowadays the indication for treatment have become relatively clear, indeed uncertainity remains for what concerns the proper treatment modality (surgical or endovascular)in terms of both the risk and the mid and long-term efficacy of the two procedures. The ''Unruptured Aneurysms Italian Study'' is a cooperative prospective study which aims to delineate the ''State of the Art'' in a nation based population. It has been designed: 1) to depict the nationwide modality of treatment of Unruptured Aneurysms, 2) to assess in the most objective way the overall treatment-related mortality and morbidity as well as the surgical and endovascular risk in the respective patient populations (it is not a surgical versus endovascular study) and 3) to asses the efficacy of the different procedures in the mid and long term periods. The study started on June 2003 and to June 2006, 637 patients have been enrolled. The study will end when the 1000th patient is enrolled. PMID- 17108887 TI - The guidelines for the diagnosis and treatment of spasticity. AB - Spasticity is a predominant clinical sign appearing in different neurological diseases. It is always flanked by various degrees of muscle weakness. The clinical evaluation of a spastic patient is score according to varius internationally approved evaluation scales (Ashworth scale, muscle spasms scale, and FIM disability scale). The treatment of spasticity is mostly a symptomatic treatment aimed to relief muscle hypertonus thus increasing both motor performance and improving nursing. Many molecules are frequently being used orally with poor results or with the onset of undesired side effects. In fact oral baclofen, diazepam and tizanidine often have poor effect on spasticity and bring frequently to the appearance of undesired side effects caused by the concentration of these molecule at the brain level. Intrathecal baclofen is a good option to treat diffuse spasticity through the infusion of baclofen into the spinal CSF space. When baclofen is administered intrathecally at the spinal level it distributes with a concentration-gradient between caudal and rostral level of the spine that was calculated as 4:1 thus avoiding its concentration at the brain level when given at a therapeutical dosage. This fact avoids any undesired side effect due to the action of baclofen at the brain level. Botulinum toxin as well as peripheral neurotomies are very helpful in those cases in whom spasticity is mainly restricted to few muscular groups. A correct flow-chart to diagnose and treat the patient is mandatory to achieve the best results for each patient according to his spasticity and residual motor ability. PMID- 17108888 TI - Upper eyelid gold weight implants in patients with facial nerve palsy. Surgical technique. AB - Facial nerve injuries produce lagophthalmos and consequent ocular disease caused by corneal exposure. The management of the affected eye in patients with facial palsy has been improved. Previously ointment, eye drops, taping, partial or complete tarsorrhaphy was the primary treatment of the inability to close the eyelid. Other mechanical techniques for reanimating lid closure, including palpebral springs, encircling the upper and lower eyelids with silicone or fascia lata, and temporalis muscle transfer. The most popular and widely used static procedure in facial nerve palsy is the upper eyelid gold weight implant. This procedure is the goal of the treatment for the restoration of function and cosmesis to the paralyzed eyelids. The surgical technique used for lid load insertion are described below. PMID- 17108889 TI - Aggressive eosinophilic granuloma of the parietal bone. An immunohystochemical study of Ki-67 expression. AB - Solitary eosinophilic granuloma (EG) of the skull is a rare lesion, the natural history of which is still to be defined. We report a case of a 26-year-old female who presented with progressive headache and nausea accompanied by a painful firm mass in her left parietal region, which grew very rapidly during the last two weeks before admission. Computed tomography scan showed an osteolytic lesion, which on magnetic resonance imaging appeared hyperintense on both T1- and T2 weighted images, with marked and heterogeneous enhancement after gadolinium administration. Total surgical excision of the lesion was performed and histopathological diagnosis was compatible with eosinophilic granuloma. Immuno histochemical study of Ki-67 antigen expression was also performed with a labelling index of 10%. In a review of the pertinent literature, we found one case report showing a Ki-67 labelling index of 6.2% in a patient harboring EG of the occipital bone. These two relatively high percentages of proliferative activity suggest a role of local Langerhans'cell proliferation, along with that of inflammatory response, in the aggressive clinical course and rapid expansion observed in some rare cases of solitary eosinophilic granuloma. PMID- 17108890 TI - Fatalities and injuries from falls among older adults--United States, 1993-2003 and 2001-2005. AB - Unintentional falls are a common occurrence among older adults, affecting approximately 30% of persons aged > or =65 years each year. The injuries received from a fall can result in death, disability, nursing-home admission, and direct medical costs. In 2003, a total of 13,700 persons aged > or =65 years died from falls, and 1.8 million were treated in emergency departments (EDs) for nonfatal injuries from falls. Falls cause the majority of hip fractures, which often result in long-term functional impairments that might require admission to a nursing home for a year or more. To examine trends in fatal and nonfatal falls among older persons, CDC analyzed U.S. rates of 1) fatalities from falls (during 1993-2003), 2) hospitalizations for hip fractures (1993-2003), and 3) nonfatal injuries resulting from falls in persons treated in EDs (2001-2005). This report summarizes the results of those analyses, which indicated that, during 1993-2003, the overall rate of fatal falls among persons aged > or =65 years increased, and the rate of hospitalizations for hip fractures decreased; during 2001-2005, the change in the overall rate of nonfatal injuries from falls was not statistically significant. However, disparities by sex existed for all three measures. Certain interventions can reduce falls (e.g., exercising regularly or having medicines reviewed to reduce side effects and interactions), but implementation at the community level remains limited, and additional measures are needed to promote widespread adoption. PMID- 17108891 TI - Self-rated fair or poor health among adults with diabetes--United States, 1996 2005. AB - Diabetes mellitus affects nearly 21 million persons in the United States. Maintaining and improving health-related quality of life among persons with diabetes is a public health goal. Healthy People 2010 includes self-rated health as one of three surveillance tools that can be used to measure health-related quality of life. To assess the prevalence of self-rated fair or poor health among U.S. adults with diabetes and to identify factors associated with fair or poor health, CDC analyzed 1996-2005 Behavioral Risk Factor Surveillance System (BRFSS) data. This report summarizes the findings of that analysis, which indicated that self-rated fair or poor health was three times more common among adults with diabetes than among those without diabetes and that the prevalence increased during 1996-2005 among young adults (i.e., aged 18-44 years) with diabetes. The results underscore the need for 1) continued interventions to promote healthy behaviors and prevent diabetes and 2) interventions for persons with diabetes to help them better manage their diabetes and prevent diabetes complications, which can increase their perceived quality of life. PMID- 17108892 TI - Brief report: hazardous materials release resulting from home production of biodiesel--Colorado, May 2006. AB - On May 7, 2006, a hazardous materials (HazMat) release occurred in a residential area of Colorado when a homeowner who was processing a tank of homemade biodiesel fuel forgot to turn off the tank's heating element and left for the weekend. The heating element overheated and caused a fire that burned the surrounding shed and equipment. The shed had contained >600 gallons of biodiesel and recycled restaurant cooking oil, smaller amounts of glycerin and sodium hydroxide, and 1 gallon containers of sulfuric and phosphoric acid; a mixture of these ingredients seeped into the ground during the fire. A certified HazMat team and the local fire department responded. Investigators found seven 55-gallon barrels of methanol and other hazardous materials outside the shed. No injuries or evacuations occurred. To prevent potential injuries, biodiesel should be purchased from a licensed commercial source. PMID- 17108893 TI - Reverse dosimetry: interpreting trihalomethanes biomonitoring data using physiologically based pharmacokinetic modeling. AB - Biomonitoring data provide evidence of exposure of environmental chemicals but are not, by themselves, direct measures of exposure. To use biomonitoring data in understanding exposure, physiologically based pharmacokinetic (PBPK) modeling can be used in a reverse dosimetry approach to assess a distribution of exposures possibly associated with specific blood or urine levels of compounds. Reverse dosimetry integrates PBPK modeling with exposure pattern characterization, Monte Carlo analysis, and statistical tools to estimate a distribution of exposures that are consistent with biomonitoring data in a population. The present study used an existing PBPK model for chloroform as a generic framework to develop PBPK models for other trihalomethanes (THMs). Using Monte Carlo sampling techniques, probabilistic information about pharmacokinetics and exposure patterns was included to estimate distributions of THMs concentrations in blood in relation to various exposure patterns in a diverse population. In addition, the possibility of inhibition of hepatic metabolism among THMs was evaluated under the scenarios of household exposure. These studies demonstrated how PBPK modeling can be used as a tool to estimate a population distribution of exposures that could have resulted in particular biomonitoring results. When toxicity level is known, this tool can also be used to estimate proportion of population above levels associated with health risk. PMID- 17108894 TI - Evaluation of the recursive model approach for estimating particulate matter infiltration efficiencies using continuous light scattering data. AB - Quantifying particulate matter (PM) infiltration efficiencies (F(inf)) in individual homes is an important part of PM exposure assessment because individuals spend the majority of time indoors. While F(inf) of fine PM has most commonly been estimated using tracer species such as sulfur, here we evaluate an alternative that does not require particle collection, weighing and compositional analysis, and can be applied in situations with indoor sources of sulfur, such as environmental tobacco smoke, gas pilot lights, and humidifier use. This alternative method involves applying a recursive mass balance model (recursive model, RM) to continuous indoor and outdoor concentration measurements (e.g., light scattering data from nephelometers). We show that the RM can reliably estimate F(inf), a crucial parameter for determining exposure to particles of outdoor origin. The RM F(inf) estimates showed good agreement with the conventional filter-based sulfur tracer approach. Our simulation results suggest that the RM F(inf) estimates are minimally impacted by measurement error. In addition, the average light scattering response per unit mass concentration was greater indoors than outdoors; after correcting for differences in light scattering response the median deviation from sulfur F(inf) was reduced from 15 to 11%. Thus, we have verified the RM applied to light scattering data. We show that the RM method is unable to provide satisfactory estimates of the individual components of F(inf) (penetration efficiency, air exchange rate, and deposition rate). However, this approach may allow F(inf) to be estimated in more residences, including those with indoor sources of sulfur. We show that individual homes vary in their infiltration efficiencies, thereby contributing to exposure misclassification in epidemiological studies that assign exposures using ambient monitoring data. This variation across homes indicates the need for home specific estimation methods, such as the RM or sulfur tracer, instead of techniques that give average estimates of infiltration across homes. PMID- 17108895 TI - Daily mortality and particulate matter in different size classes in Erfurt, Germany. AB - The link between elevated concentrations of ambient particulate matter (PM) and increased mortality has been investigated in numerous studies. Here we analyzed the role of different particle size fractions with respect to total and cardio respiratory mortality in Erfurt, Germany, between 1995 and 2001. Number concentrations (NC) of PM were measured using an aerosol spectrometer consisting of a Differential Mobility Particle Sizer and a Laser Aerosol Spectrometer to characterize particles between 0.01 and 0.5 and between 0.1 and 2.5 microm, respectively. We derived daily means of particle NC for ultrafine (0.01-0.1 microm) and for fine particles (0.01-2.5 microm). Assuming spherical particles of a constant density, we estimated the mass concentrations (MC) of particles in these size ranges. Concurrently, data on daily total and cardio-respiratory death counts were obtained from local health authorities. The data were analyzed using Poisson Generalized Additive Models adjusting for trend, seasonality, influenza epidemics, day of the week, and meteorology using smooth functions or indicator variables. We found statistically significant associations between elevated ultrafine particle (UFP; diameter: 0.01-0.1 microm) NC and total as well as cardio-respiratory mortality, each with a 4 days lag. The relative mortality risk (RR) for a 9748 cm(-3) increase in UFP NC was RR=1.029 and its 95% confidence interval (CI)=1.003-1.055 for total mortality. For cardio-respiratory mortality we found: RR=1.031, 95% CI: 1.003-1.060. No association between fine particle MC and mortality was found. This study shows that UFP, representing fresh combustion particles, may be an important component of urban air pollution associated with health effects. PMID- 17108896 TI - Becoming an advocate. PMID- 17108900 TI - Stress and the skin: a meeting report of the Weill Cornell Symposium on the Science of Dermatology. PMID- 17108902 TI - Healing slack skin. AB - A non-lethal, murine knockout of fibulin-5 with features of autosomal recessive cutis laxa and marked defects in elastic fiber formation amplifies previous observations on the minimal role of the elastic component in acute, cutaneous wound healing. More demanding wound models or long-term studies may yet reveal how fibulin-5 and elastin impact the quality of repair. PMID- 17108903 TI - Oxidative stress in the pathogenesis of skin disease. AB - Skin is the largest body organ that serves as an important environmental interface providing a protective envelope that is crucial for homeostasis. On the other hand, the skin is a major target for toxic insult by a broad spectrum of physical (i.e. UV radiation) and chemical (xenobiotic) agents that are capable of altering its structure and function. Many environmental pollutants are either themselves oxidants or catalyze the production of reactive oxygen species (ROS) directly or indirectly. ROS are believed to activate proliferative and cell survival signaling that can alter apoptotic pathways that may be involved in the pathogenesis of a number of skin disorders including photosensitivity diseases and some types of cutaneous malignancy. ROS act largely by driving several important molecular pathways that play important roles in diverse pathologic processes including ischemia-reperfusion injury, atherosclerosis, and inflammatory responses. The skin possesses an array of defense mechanisms that interact with toxicants to obviate their deleterious effect. These include non enzymatic and enzymatic molecules that function as potent antioxidants or oxidant degrading systems. Unfortunately, these homeostatic defenses, although highly effective, have limited capacity and can be overwhelmed thereby leading to increased ROS in the skin that can foster the development of dermatological diseases. One approach to preventing or treating these ROS-mediated disorders is based on the administration of various antioxidants in an effort to restore homeostasis. Although many antioxidants have shown substantive efficacy in cell culture systems and in animal models of oxidant injury, unequivocal confirmation of their beneficial effects in human populations has proven elusive. PMID- 17108917 TI - Order for microbes. PMID- 17108904 TI - Computer simulation of native epidermal enzyme structures in the presence and absence of hydrogen peroxide (H2O2): potential and pitfalls. AB - The human epidermis is especially vulnerable to oxidative stress, which in turn leads to oxidation of important antioxidant enzymes, other proteins, and peptides. Molecular dynamic computer modelling is a new powerful tool to predict or confirm oxidative stress-mediated structural changes consequently altering the function of enzymes/proteins/peptides. Here we used examples of important epidermal antioxidant enzymes before and after hydrogen peroxide (H(2)O(2)) mediated oxidation of susceptible amino-acid residues (i.e. tryptophan, methionine, cysteine, and selenocysteine), which can affect enzyme active sites, cofactor binding, or dimerization/tetramerization domains. Computer modelling predicts that enzyme active sites are altered by H(2)O(2)-mediated oxidation in thioredoxin reductase (TR) and acetylcholinesterase (AchE), whereas cofactor nicotinamide adenine dinucleotide phosphate (reduced form) binding is affected in both catalase and TR but not in glutathione peroxidase. Dimerization is prevented in catalase. These structural changes lead to impaired functionality. Fourier transform-Raman- and Fluorescence spectroscopy together with enzyme kinetics support the results. There are limitations of modelling as demonstrated on the AchE substrate-binding domain, where the computer predicted deactivation, which could not be confirmed by enzyme kinetics. Computer modelling coupled with classical biochemical techniques offers a new powerful tool in cutaneous biology to explore oxidative stress-mediated metabolic changes in the skin. PMID- 17108918 TI - Congressional agendas. PMID- 17108919 TI - Success and successor. PMID- 17108921 TI - The politics of breathing. PMID- 17108922 TI - WHO boss faces test of independence. PMID- 17108924 TI - Q and A: Bart Gordon. PMID- 17108925 TI - Gunmen seize academics at Baghdad ministry. PMID- 17108926 TI - UK civil servants accused of warping science. PMID- 17108927 TI - German stem-cell law under fire. PMID- 17108928 TI - Neanderthal genome sees first light. PMID- 17108930 TI - The chips are down. PMID- 17108933 TI - Genetic information: codes and enigmas. PMID- 17108934 TI - Waste management: one man's trash... PMID- 17108935 TI - Charmed, I'm sure. PMID- 17108936 TI - Creationism, evolution: nothing has been proved. PMID- 17108937 TI - Creationists attack secular education in Russia. PMID- 17108938 TI - Criticism: what to do about science's bad public image? PMID- 17108939 TI - Answering critics can add fuel to controversy. PMID- 17108940 TI - Safe handling of nanotechnology. PMID- 17108944 TI - Evolutionary biology: ancient genomics is born. PMID- 17108945 TI - Seismology: greatness thrust upon them. PMID- 17108947 TI - Fluid dynamics: spinning discs in the lab. PMID- 17108948 TI - Environmental chemistry: browning the waters. PMID- 17108949 TI - Upper Palaeolithic infant burials. AB - Decorations on the bodies of newborns indicate that they were probably important in their community. Several adult graves from the Stone Age (Upper Palaeolithic period) have been found but child burials seem to be rare, which has prompted discussion about whether this apparently different treatment of infants could be significant. Here we describe two recently discovered infant burials from this period at Krems-Wachtberg in Lower Austria, in which the bodies were covered with red ochre and decorated with ornaments and were therefore probably ritually buried. These findings indicate that even newborns were considered to be full members of these hunter-gatherer communities about 27,000 years ago. PMID- 17108950 TI - Materials: carbon nanotubes in an ancient Damascus sabre. AB - The steel of Damascus blades, which were first encountered by the Crusaders when fighting against Muslims, had features not found in European steels--a characteristic wavy banding pattern known as damask, extraordinary mechanical properties, and an exceptionally sharp cutting edge. Here we use high-resolution transmission electron microscopy to examine a sample of Damascus sabre steel from the seventeenth century and find that it contains carbon nanotubes as well as cementite nanowires. This microstructure may offer insight into the beautiful banding pattern of the ultrahigh-carbon steel created from an ancient recipe that was lost long ago. PMID- 17108952 TI - The receptors and cells for mammalian taste. AB - The emerging picture of taste coding at the periphery is one of elegant simplicity. Contrary to what was generally believed, it is now clear that distinct cell types expressing unique receptors are tuned to detect each of the five basic tastes: sweet, sour, bitter, salty and umami. Importantly, receptor cells for each taste quality function as dedicated sensors wired to elicit stereotypic responses. PMID- 17108953 TI - Comparative chemosensation from receptors to ecology. AB - Odour perception is initiated by specific interactions between odorants and a large repertoire of receptors in olfactory neurons. During the past few years, considerable progress has been made in tracing olfactory perception from the odorant receptor protein to the activity of olfactory neurons to higher processing centres and, ultimately, to behaviour. The most complete picture is emerging for the simplest olfactory system studied--that of the fruitfly Drosophila melanogaster. Comparison of rodent, insect and nematode olfaction reveals surprising differences and unexpected similarities among chemosensory systems. PMID- 17108954 TI - Insects as chemosensors of humans and crops. AB - Insects transmit disease to hundreds of millions of people a year, and cause enormous losses to the world's agricultural output. Many insects find the human or plant hosts on which they feed, and identify and locate their mates, primarily through olfaction and taste. Major advances have recently been made in understanding insect chemosensation at the molecular and cellular levels. These advances have provided new opportunities to control insects that cause massive damage to health and agriculture across the world. PMID- 17108955 TI - Pheromonal communication in vertebrates. AB - Recent insights have revolutionized our understanding of the importance of chemical signals in influencing vertebrate behaviour. Previously unknown families of pheromonal signals have been identified that are expanding the traditional definition of a pheromone. Although previously regarded as functioning independently, the main olfactory and vomeronasal systems have been found to have considerable overlap in terms of the chemosignals they detect and the effects that they mediate. Studies using gene-targeted mice have revealed an unexpected diversity of chemosensory systems and their underlying cellular and molecular mechanisms. Future developments could show how the functions of the different chemosensory systems are integrated to regulate innate and learned behavioural and physiological responses to pheromones. PMID- 17108956 TI - Smell images and the flavour system in the human brain. AB - Flavour perception is one of the most complex of human behaviours. It involves almost all of the senses, particularly the sense of smell, which is involved through odour images generated in the olfactory pathway. In the human brain, the perceptual systems are closely linked to systems for learning, memory, emotion and language, so distributed neural mechanisms contribute to food preference and food cravings. Greater recognition of the role of the brain's flavour system and its connection with eating behaviour is needed for a deeper understanding of why people eat what they do, and to generate better recommendations about diet and nutrition. PMID- 17108957 TI - The plant immune system. AB - Many plant-associated microbes are pathogens that impair plant growth and reproduction. Plants respond to infection using a two-branched innate immune system. The first branch recognizes and responds to molecules common to many classes of microbes, including non-pathogens. The second responds to pathogen virulence factors, either directly or through their effects on host targets. These plant immune systems, and the pathogen molecules to which they respond, provide extraordinary insights into molecular recognition, cell biology and evolution across biological kingdoms. A detailed understanding of plant immune function will underpin crop improvement for food, fibre and biofuels production. PMID- 17108958 TI - Analysis of one million base pairs of Neanderthal DNA. AB - Neanderthals are the extinct hominid group most closely related to contemporary humans, so their genome offers a unique opportunity to identify genetic changes specific to anatomically fully modern humans. We have identified a 38,000-year old Neanderthal fossil that is exceptionally free of contamination from modern human DNA. Direct high-throughput sequencing of a DNA extract from this fossil has thus far yielded over one million base pairs of hominoid nuclear DNA sequences. Comparison with the human and chimpanzee genomes reveals that modern human and Neanderthal DNA sequences diverged on average about 500,000 years ago. Existing technology and fossil resources are now sufficient to initiate a Neanderthal genome-sequencing effort. PMID- 17108959 TI - Hydrodynamic turbulence cannot transport angular momentum effectively in astrophysical disks. AB - The most efficient energy sources known in the Universe are accretion disks. Those around black holes convert 5-40 per cent of rest-mass energy to radiation. Like water circling a drain, inflowing mass must lose angular momentum, presumably by vigorous turbulence in disks, which are essentially inviscid. The origin of the turbulence is unclear. Hot disks of electrically conducting plasma can become turbulent by way of the linear magnetorotational instability. Cool disks, such as the planet-forming disks of protostars, may be too poorly ionized for the magnetorotational instability to occur, and therefore essentially unmagnetized and linearly stable. Nonlinear hydrodynamic instability often occurs in linearly stable flows (for example, pipe flows) at sufficiently large Reynolds numbers. Although planet-forming disks have extreme Reynolds numbers, keplerian rotation enhances their linear hydrodynamic stability, so the question of whether they can be turbulent and thereby transport angular momentum effectively is controversial. Here we report a laboratory experiment, demonstrating that non magnetic quasi-keplerian flows at Reynolds numbers up to millions are essentially steady. Scaled to accretion disks, rates of angular momentum transport lie far below astrophysical requirements. By ruling out purely hydrodynamic turbulence, our results indirectly support the magnetorotational instability as the likely cause of turbulence, even in cool disks. PMID- 17108960 TI - Half-metallic graphene nanoribbons. AB - Electrical current can be completely spin polarized in a class of materials known as half-metals, as a result of the coexistence of metallic nature for electrons with one spin orientation and insulating nature for electrons with the other. Such asymmetric electronic states for the different spins have been predicted for some ferromagnetic metals--for example, the Heusler compounds--and were first observed in a manganese perovskite. In view of the potential for use of this property in realizing spin-based electronics, substantial efforts have been made to search for half-metallic materials. However, organic materials have hardly been investigated in this context even though carbon-based nanostructures hold significant promise for future electronic devices. Here we predict half metallicity in nanometre-scale graphene ribbons by using first-principles calculations. We show that this phenomenon is realizable if in-plane homogeneous electric fields are applied across the zigzag-shaped edges of the graphene nanoribbons, and that their magnetic properties can be controlled by the external electric fields. The results are not only of scientific interest in the interplay between electric fields and electronic spin degree of freedom in solids but may also open a new path to explore spintronics at the nanometre scale, based on graphene. PMID- 17108961 TI - Free-electron-like dispersion in an organic monolayer film on a metal substrate. AB - Thin films of molecular organic semiconductors are attracting much interest for use in electronic and optoelectronic applications. The electronic properties of these materials and their interfaces are therefore worth investigating intensively, particularly the degree of electron delocalization that can be achieved. If the delocalization is appreciable, it should be accompanied by an observable electronic band dispersion. But so far only limited experimental data on the intermolecular dispersion of electronic states in molecular materials is available, and the mechanism(s) of electron delocalization in molecular materials are also not well understood. Here we report scanning tunnelling spectroscopy observations of an organic monolayer film on a silver substrate, revealing a completely delocalized two-dimensional band state that is characterized by a metal-like parabolic dispersion with an effective mass of m* = 0.47m(e), where m(e) is the bare electron mass. This dispersion is far stronger than expected for the organic film alone, and arises as a result of strong substrate-mediated coupling between the molecules within the monolayer. PMID- 17108962 TI - Flushing submarine canyons. AB - The continental slope is a steep, narrow fringe separating the coastal zone from the deep ocean. During low sea-level stands, slides and dense, sediment-laden flows erode the outer continental shelf and the continental slope, leading to the formation of submarine canyons that funnel large volumes of sediment and organic matter from shallow regions to the deep ocean(1). During high sea-level stands, such as at present, these canyons still experience occasional sediment gravity flows(2-5), which are usually thought to be triggered by sediment failure or river flooding. Here we present observations from a submarine canyon on the Gulf of Lions margin, in the northwest Mediterranean Sea, that demonstrate that these flows can also be triggered by dense shelf water cascading (DSWC)-a type of current that is driven solely by seawater density contrast. Our results show that DSWC can transport large amounts of water and sediment, reshape submarine canyon floors and rapidly affect the deep-sea environment. This cascading is seasonal, resulting from the formation of dense water by cooling and/or evaporation, and occurs on both high- and low-latitude continental margins(6-8). DSWC may therefore transport large amounts of sediment and organic matter to the deep ocean. Furthermore, changes in the frequency and intensity of DSWC driven by future climate change may have a significant impact on the supply of organic matter to deep-sea ecosystems and on the amount of carbon stored on continental margins and in ocean basins. PMID- 17108963 TI - Predicting the endpoints of earthquake ruptures. AB - The active fault traces on which earthquakes occur are generally not continuous, and are commonly composed of segments that are separated by discontinuities that appear as steps in map-view. Stress concentrations resulting from slip at such discontinuities may slow or stop rupture propagation and hence play a controlling role in limiting the length of earthquake rupture. Here I examine the mapped surface rupture traces of 22 historical strike-slip earthquakes with rupture lengths ranging between 10 and 420 km. I show that about two-thirds of the endpoints of strike-slip earthquake ruptures are associated with fault steps or the termini of active fault traces, and that there exists a limiting dimension of fault step (3-4 km) above which earthquake ruptures do not propagate and below which rupture propagation ceases only about 40 per cent of the time. The results are of practical importance to seismic hazard analysis where effort is spent attempting to place limits on the probable length of future earthquakes on mapped active faults. Physical insight to the dynamics of the earthquake rupture process is further gained with the observation that the limiting dimension appears to be largely independent of the earthquake rupture length. It follows that the magnitude of stress changes and the volume affected by those stress changes at the driving edge of laterally propagating ruptures are largely similar and invariable during the rupture process regardless of the distance an event has propagated or will propagate. PMID- 17108964 TI - Allee effects and pulsed invasion by the gypsy moth. AB - Biological invasions pose considerable threats to the world's ecosystems and cause substantial economic losses. A prime example is the invasion of the gypsy moth in the United States, for which more than $194 million was spent on management and monitoring between 1985 and 2004 alone. The spread of the gypsy moth across eastern North America is, perhaps, the most thoroughly studied biological invasion in the world, providing a unique opportunity to explore spatiotemporal variability in rates of spread. Here we describe evidence for periodic pulsed invasions, defined as regularly punctuated range expansions interspersed among periods of range stasis. We use a theoretical model with parameter values estimated from long-term monitoring data to show how an interaction between strong Allee effects (negative population growth at low densities) and stratified diffusion (most individuals disperse locally, but a few seed new colonies by long-range movement) can explain the invasion pulses. Our results indicate that suppressing population peaks along range borders might greatly slow invasion. PMID- 17108965 TI - Haemagglutinin mutations responsible for the binding of H5N1 influenza A viruses to human-type receptors. AB - H5N1 influenza A viruses have spread to numerous countries in Asia, Europe and Africa, infecting not only large numbers of poultry, but also an increasing number of humans, often with lethal effects. Human and avian influenza A viruses differ in their recognition of host cell receptors: the former preferentially recognize receptors with saccharides terminating in sialic acid-alpha2,6 galactose (SAalpha2,6Gal), whereas the latter prefer those ending in SAalpha2,3Gal (refs 3-6). A conversion from SAalpha2,3Gal to SAalpha2,6Gal recognition is thought to be one of the changes that must occur before avian influenza viruses can replicate efficiently in humans and acquire the potential to cause a pandemic. By identifying mutations in the receptor-binding haemagglutinin (HA) molecule that would enable avian H5N1 viruses to recognize human-type host cell receptors, it may be possible to predict (and thus to increase preparedness for) the emergence of pandemic viruses. Here we show that some H5N1 viruses isolated from humans can bind to both human and avian receptors, in contrast to those isolated from chickens and ducks, which recognize the avian receptors exclusively. Mutations at positions 182 and 192 independently convert the HAs of H5N1 viruses known to recognize the avian receptor to ones that recognize the human receptor. Analysis of the crystal structure of the HA from an H5N1 virus used in our genetic experiments shows that the locations of these amino acids in the HA molecule are compatible with an effect on receptor binding. The amino acid changes that we identify might serve as molecular markers for assessing the pandemic potential of H5N1 field isolates. PMID- 17108966 TI - Bacterial chromatin organization by H-NS protein unravelled using dual DNA manipulation. AB - Both prokaryotic and eukaryotic organisms contain DNA bridging proteins, which can have regulatory or architectural functions. The molecular and mechanical details of such proteins are hard to obtain, in particular if they involve non specific interactions. The bacterial nucleoid consists of hundreds of DNA loops, shaped in part by non-specific DNA bridging proteins such as histone-like nucleoid structuring protein (H-NS), leucine-responsive regulatory protein (Lrp) and SMC (structural maintenance of chromosomes) proteins. We have developed an optical tweezers instrument that can independently handle two DNA molecules, which allows the systematic investigation of protein-mediated DNA-DNA interactions. Here we use this technique to investigate the abundant non-specific nucleoid-associated protein H-NS, and show that H-NS is dynamically organized between two DNA molecules in register with their helical pitch. Our optical tweezers also allow us to carry out dynamic force spectroscopy on non-specific DNA binding proteins and thereby to determine an energy landscape for the H-NS DNA interaction. Our results explain how the bacterial nucleoid can be effectively compacted and organized, but be dynamic in nature and accessible to DNA-tracking motor enzymes. Finally, our experimental approach is widely applicable to other DNA bridging proteins, as well as to complex DNA interactions involving multiple DNA molecules. PMID- 17108968 TI - Stem-cell biology: a move in the right direction. PMID- 17108969 TI - Thymosin beta4 induces adult epicardial progenitor mobilization and neovascularization. AB - Cardiac failure has a principal underlying aetiology of ischaemic damage arising from vascular insufficiency. Molecules that regulate collateral growth in the ischaemic heart also regulate coronary vasculature formation during embryogenesis. Here we identify thymosin beta4 (Tbeta4) as essential for all aspects of coronary vessel development in mice, and demonstrate that Tbeta4 stimulates significant outgrowth from quiescent adult epicardial explants, restoring pluripotency and triggering differentiation of fibroblasts, smooth muscle cells and endothelial cells. Tbeta4 knockdown in the heart is accompanied by significant reduction in the pro-angiogenic cleavage product N-acetyl-seryl aspartyl-lysyl-proline (AcSDKP). Although injection of AcSDKP was unable to rescue Tbeta4 mutant hearts, it significantly enhanced endothelial cell differentiation from adult epicardially derived precursor cells. This study identifies Tbeta4 and AcSDKP as potent stimulators of coronary vasculogenesis and angiogenesis, and reveals Tbeta4-induced adult epicardial cells as a viable source of vascular progenitors for continued renewal of regressed vessels at low basal level or sustained neovascularization following cardiac injury. PMID- 17108970 TI - Endogenous neurosteroids regulate GABAA receptors through two discrete transmembrane sites. AB - Inhibitory neurotransmission mediated by GABA(A) receptors can be modulated by the endogenous neurosteroids, allopregnanolone and tetrahydro deoxycorticosterone. Neurosteroids are synthesized de novo in the brain during stress, pregnancyand after ethanol consumption, and disrupted steroid regulation of GABAergic transmission is strongly implicated in several debilitating conditions such as panic disorder, major depression, schizophrenia, alcohol dependence and catamenial epilepsy. Determining how neurosteroids interact with the GABA(A) receptor is a prerequisite for understanding their physiological and pathophysiological roles in the brain. Here we identify two discrete binding sites in the receptor's transmembrane domains that mediate the potentiating and direct activation effects of neurosteroids. They potentiate GABA responses from a cavity formed by the alpha-subunit transmembrane domains, whereas direct receptor activation is initiated by interfacial residues between alpha and beta subunits and is enhanced by steroid binding to the potentiation site. Thus, significant receptor activation by neurosteroids relies on occupancy of both the activation and potentiation sites. These sites are highly conserved throughout the GABA(A )receptor family, and their identification provides a unique opportunity for the development of new therapeutic, neurosteroid-based ligands and transgenic disease models of neurosteroid dysfunction. PMID- 17108971 TI - Repression of p53 activity by Smyd2-mediated methylation. AB - Specific sites of lysine methylation on histones correlate with either activation or repression of transcription. The tumour suppressor p53 (refs 4-7) is one of only a few non-histone proteins known to be regulated by lysine methylation. Here we report a lysine methyltransferase, Smyd2, that methylates a previously unidentified site, Lys 370, in p53. This methylation site, in contrast to the known site Lys 372, is repressing to p53-mediated transcriptional regulation. Smyd2 helps to maintain low concentrations of promoter-associated p53. We show that reducing Smyd2 concentrations by short interfering RNA enhances p53-mediated apoptosis. We find that Set9-mediated methylation of Lys 372 inhibits Smyd2 mediated methylation of Lys 370, providing regulatory cross-talk between post translational modifications. In addition, we show that the inhibitory effect of Lys 372 methylation on Lys 370 methylation is caused, in part, by blocking the interaction between p53 and Smyd2. Thus, similar to histones, p53 is subject to both activating and repressing lysine methylation. Our results also predict that Smyd2 may function as a putative oncogene by methylating p53 and repressing its tumour suppressive function. PMID- 17108974 TI - Frontiers in drug discovery. PMID- 17108972 TI - Mesoangioblast stem cells ameliorate muscle function in dystrophic dogs. AB - Duchenne muscular dystrophy remains an untreatable genetic disease that severely limits motility and life expectancy in affected children. The only animal model specifically reproducing the alterations in the dystrophin gene and the full spectrum of human pathology is the golden retriever dog model. Affected animals present a single mutation in intron 6, resulting in complete absence of the dystrophin protein, and early and severe muscle degeneration with nearly complete loss of motility and walking ability. Death usually occurs at about 1 year of age as a result of failure of respiratory muscles. Here we report that intra-arterial delivery of wild-type canine mesoangioblasts (vessel-associated stem cells) results in an extensive recovery of dystrophin expression, normal muscle morphology and function (confirmed by measurement of contraction force on single fibres). The outcome is a remarkable clinical amelioration and preservation of active motility. These data qualify mesoangioblasts as candidates for future stem cell therapy for Duchenne patients. PMID- 17108975 TI - Smart drug discovery leveraging innovative technologies and predictive knowledge. PMID- 17108976 TI - Drug discovery through industry-academic partnerships. PMID- 17108977 TI - Stuart Levy. PMID- 17108978 TI - Marlene Haffner. PMID- 17108979 TI - Puzzling through fragment-based drug design. PMID- 17108980 TI - A sweet success for substrate engineering. PMID- 17108981 TI - Combinatorial biosynthesis in symbiotic bacteria. PMID- 17108982 TI - The connectivity map. PMID- 17108984 TI - Developing a new resource for drug discovery: marine actinomycete bacteria. PMID- 17108985 TI - Production of isoprenoid pharmaceuticals by engineered microbes. AB - Throughout human history, natural products have been the foundation for the discovery and development of therapeutics used to treat diseases ranging from cardiovascular disease to cancer. Their chemical diversity and complexity have provided structural scaffolds for small-molecule drugs and have consistently served as inspiration for medicinal design. However, the chemical complexity of natural products also presents one of the main roadblocks for production of these pharmaceuticals on an industrial scale. Chemical synthesis of natural products is often difficult and expensive, and isolation from their natural sources is also typically low yielding. Synthetic biology and metabolic engineering offer an alternative approach that is becoming more accessible as the tools for engineering microbes are further developed. By reconstructing heterologous metabolic pathways in genetically tractable host organisms, complex natural products can be produced from inexpensive sugar starting materials through large scale fermentation processes. In this Perspective, we discuss ongoing research aimed toward the production of terpenoid natural products in genetically engineered Escherichia coli and Saccharomyces cerevisiae. PMID- 17108986 TI - Synthetic therapeutic antibodies. PMID- 17108987 TI - New approaches to molecular cancer therapeutics. AB - Cancer drug development is leading the way in exploiting molecular biological and genetic information to develop "personalized" medicine. The new paradigm is to develop agents that target the precise molecular pathology driving the progression of individual cancers. Drug developers have benefited from decades of academic cancer research and from investment in genomics, genetics and automation; their success is exemplified by high-profile drugs such as Herceptin (trastuzumab), Gleevec (imatinib), Tarceva (erlotinib) and Avastin (bevacizumab). However, only 5% of cancer drugs entering clinical trials reach marketing approval. Cancer remains a high unmet medical need, and many potential cancer targets remain undrugged. In this review we assess the status of the discovery and development of small-molecule cancer therapeutics. We show how chemical biology approaches offer techniques for interconnecting elements of the traditional linear progression from gene to drug, thereby providing a basis for increasing speed and success in cancer drug discovery. PMID- 17108988 TI - Drug discovery and development for neglected parasitic diseases. AB - Diseases caused by tropical parasites affect hundreds of millions of people worldwide but have been largely neglected for drug development because they affect poor people in poor regions of the world. Most of the current drugs used to treat these diseases are decades old and have many limitations, including the emergence of drug resistance. This review will summarize efforts to reinvigorate the drug development pipeline for these diseases, which is driven in large part by support from major philanthropies. The organisms responsible for these diseases have a fascinating biology, and many potential biochemical targets are now apparent. These neglected diseases present unique challenges to drug development that are being addressed by new consortia of scientists from academia and industry. PMID- 17108989 TI - RNAi therapeutics: a potential new class of pharmaceutical drugs. AB - The rapid identification of highly specific and potent drug candidates continues to be a substantial challenge with traditional pharmaceutical approaches. Moreover, many targets have proven to be intractable to traditional small molecule and protein approaches. Therapeutics based on RNA interference (RNAi) offer a powerful method for rapidly identifying specific and potent inhibitors of disease targets from all molecular classes. Numerous proof-of-concept studies in animal models of human disease demonstrate the broad potential application of RNAi therapeutics. The major challenge for successful drug development is identifying delivery strategies that can be translated to the clinic. With advances in this area and the commencement of multiple clinical trials with RNAi therapeutic candidates, a transformation in modern medicine may soon be realized. PMID- 17108990 TI - Identification of a novel G245R polymorphism in the IL-2 receptor beta membrane proximal domain associated with human visceral leishmaniasis. AB - Binding of the interleukin-2 (IL-2) to the IL-2 receptor (IL-2R) triggers a series of intracellular events culminating in lymphocyte proliferation and differentiation. We report here the identification of a novel G245R polymorphism in the membrane proximal domain of the IL-2 receptor beta chain (IL-2Rbeta). Present at a frequency of 7.2%, the IL-2-Rbeta G245R was identified in a population of Eastern Sudan exposed to a severe outbreak of visceral leishmaniasis (VL), a disease associated with a marked depression of T-cell antigen-specific responses. The location of the G245R polymorphism next to the box1/box2 proximal cytokine receptor homology segment and suggestive genetic association with the development of disease (P=0.043), suggest that it may affect Janus kinase (JAK) association and impair growth signal transduction. However, additional genetic association with a synonymous single nucleotide polymorphism (IL2RB+8777T) suggests that other variations of IL2RB or nearby genes participate in the highly significant linkage with VL at 22q12 previously reported for this population. PMID- 17108991 TI - The prevalence and risk factors associated with isolated untreated systolic hypertension in Korea: the Korean National Health and Nutrition Survey 2001. AB - Although isolated systolic hypertension (ISH) increases the risk of coronary heart disease and stroke, more than any other hypertension subtype, the prevalence and risk factors associated with ISH in the Korean population are not known. The 2001 Korean National Health and Nutrition Survey was a cross-sectional and nationally representative survey conducted in 2001. The prevalence of ISH by age and body mass index (BMI) was examined in 6601 Korean adults over 20 years of age. After adjusting for age, 4.32+/-0.32% of Korean adults had ISH, 5.28+/-0.37% had isolated diastolic hypertension and 5.82+/-0.36% had systolic/diastolic hypertension. The overall prevalence of ISH was found to increase directly with advancing age and increasing BMI. Although the ISH was found to be more common in men overall (4.81+/-0.50% in men, 4.12+/-0.37% in women), it was more common in women over 70 years of age. Independent variables associated with risk for ISH included advanced age, BMI, triglyceride (TG) levels, monthly income and alcohol intake. However, gender, fasting blood glucose, total cholesterol and high density lipoprotein cholesterol levels, residential area, education level and smoking were found not to be significantly associated with ISH risk. The findings of the present study demonstrate that the prevalence of untreated ISH in Korea was lower than in Western countries. Age, BMI, TG levels, monthly income and alcohol intake were associated with ISH. PMID- 17108992 TI - Gender differences in ambulatory blood pressure monitoring profile in obese, overweight and normal subjects. AB - The incidence of obesity has increased enormously in the past several decades, and has been described as a modern epidemic. Obesity is a major factor contributing to hypertension. To the best of our knowledge, no study of ambulatory blood pressure monitoring (ABPM) comparing men with women in relation to body mass indexes (BMI) has been performed. From December 2002 to May 2006, we performed 24-h ABPM in 5950 subjects (3102 men and 2848 women), with a wide range of BMI (range 15.9-53.2 kg/m(2)). We defined obese subjects as those with BMI> or =30.0 kg/m(2), overweight subjects as those with BMI>25.0 and <30.0 kg/m(2), and normal subjects as those with BMI< or =25.0 kg/m(2). Data on 989 subjects (501 men and 488 women) aged from > or =18 to < or =69 years without antihypertensive treatment, atrial fibrillation or diabetes were included for analysis. We consistently found that obese men had the expected increased heart rate compared to normal and overweight men, whereas women (normal, overweight and obese) had similar HRs. In addition, normal and obese women had similar diastolic blood pressures (BP), as opposed to obese men, who had raised diastolic BP. These results may indicate that different pathogenetic mechanisms may be involved in the relationship between obesity and hypertension in men and women. PMID- 17109003 TI - End-stage renal disease in diabetic persons: is the pandemic subsiding? AB - Analysis of data compiled by the United States Renal Data System and the National Health Interview Survey as reported in the Centers for Disease Control and Prevention's Weekly Morbidity and Mortality Report indicates that between 1990 and 2002, there has been a sharp decline in incidence rate of the number of persons with diabetes who develop end-stage renal disease. Although it is comforting to practitioners to attribute this improvement to a widely advocated regimen of renoprotection, consisting of careful regulation of hypertensive blood pressure, improved glycemic control, and lifestyle modification, evidence for this causal relationship is appearing only now. There is need to clarify the source of this epidemiologic change that will lessen the projected burden on medical and socioeconomic resources in the immediate future. PMID- 17109009 TI - Respective roles of TNF-alpha and IL-6 in the immune response-elicited by adenovirus-mediated gene transfer in mice. AB - The immunogenicity of recombinant adenoviruses (Ad) constitutes a major concern for their use in gene therapy. Antibody- and cell-mediated immune responses triggered by adenoviral vectors hamper long-term transgene expression and efficient viral readministration. We previously reported that interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha play an essential role in both the acute phase and antibody response against Ad, respectively. As TNF-alpha controls the immune response and the development of the immune system, we examined here the consequence of blockade of TNF-alpha activity through Ad-mediated gene delivery of a dimeric mouse TNFR1-IgG fusion protein on transgene expression from a second Ad. Ad encoding TNFR1-IgG (AdTNFR1-Ig) was injected intravenously along with Ad encoding beta-galactosidase or alpha1-antitrypsin transgene in wild-type (IL 6(+/+)) but also in IL-6-deficient mice (IL-6(-/-)) to analyze how TNF-alpha and IL-6 diminish liver gene transfer efficacy. Blockade of TNF-alpha leads to increased transgene expression in both wild-type and IL-6(-/-) mice due to a reduced inflammatory response and to diminished recruitment of macrophages and NK cells towards the liver. Antibody responses against adenoviral particles and expressed transgenes were only delayed in AdTNFR1-Ig-treated wild-type mice, but were markedly reduced in AdTNFR1-Ig-treated IL-6(-/-) mice. Finally, treatment of mice with etanercept, a clinically approved anti-TNF-alpha drug, confirmed the importance of controlling proinflammatory cytokines during gene therapy by adenoviral vectors. PMID- 17109010 TI - Phagocytosis of optically-trapped particles: delivery of the pure phagocytic signal. PMID- 17109011 TI - Selective degradation of the IkappaB kinase (IKK) by autophagy. PMID- 17109013 TI - Human embryonic stem cells: a resource for in vitro neuroscience research? PMID- 17109012 TI - Cancer as an evolutionary and ecological process. AB - Neoplasms are microcosms of evolution. Within a neoplasm, a mosaic of mutant cells compete for space and resources, evade predation by the immune system and can even cooperate to disperse and colonize new organs. The evolution of neoplastic cells explains both why we get cancer and why it has been so difficult to cure. The tools of evolutionary biology and ecology are providing new insights into neoplastic progression and the clinical control of cancer. PMID- 17109014 TI - An in vitro model of human dopaminergic neurons derived from embryonic stem cells: MPP+ toxicity and GDNF neuroprotection. AB - Human embryonic stem cells (hESCs) can proliferate indefinitely yet also differentiate in vitro, allowing normal human neurons to be generated in unlimited numbers. Here, we describe the development of an in vitro neurotoxicity assay using human dopaminergic neurons derived from hESCs. We showed that the dopaminergic neurotoxin 1-methyl-4-phenylpyridinium (MPP(+)), which produces features of Parkinson's disease in humans, was toxic for hESC-derived dopaminergic neurons. Treatment with glial cell line-derived neurotrophic factor protected tyrosine hydroxylase-positive neurons against MPP(+)-induced apoptotic cell death and loss of neuronal processes as well as against the formation of intracellular reactive oxygen species. The availability of human dopaminergic neurons, derived from hESCs, therefore allows for the possibility of directly examining the unique features of human dopaminergic neurons with respect to their responses to pharmacological agents as well as environmental and chemical toxins. PMID- 17109015 TI - Is mifepristone useful in psychotic depression? PMID- 17109017 TI - Assessing cognitive functioning in cannabis users: cannabis use history an important consideration. PMID- 17109021 TI - The importance of gene-environment interactions and exposure assessment in understanding human diseases. PMID- 17109024 TI - Will broad-spectrum histone deacetylase inhibitors be superseded by more specific compounds? AB - Histone deacetylase (HDAC) inhibitors can induce differentiation, cell cycle and growth arrest or in certain cases apoptosis in cancer cells. In a remarkably short period of time, especially considering that their mechanism of action remains largely undefined, HDAC inhibitors have realized both success and failure as therapeutics for cancer in clinical trials. Notably, the pleiotropic HDAC inhibitors, suberoylanilide hydroxamic acid (SAHA) and depsipeptide, have shown efficacy in a wide range of cancers, in particular for cutaneous T-cell lymphoma (CTCL), and are progressing in phase II clinical studies. However, evidence is accumulating that specific HDAC enzymes are important with respect to clinical efficacy, calling the usefulness of the classical inhibitors into question. Class I enzymes are being heralded as the most clinically relevant, however, this is still controversial and much of the information is in the private domain. Nevertheless, the potential to alter the expression of a more focused, disease related subset of genes and to limit adverse effects has prompted the development of isoform-specific HDAC inhibitors. Here, we consider the growing view that broad-spectrum HDAC inhibitors may be superseded by more specific compounds. PMID- 17109025 TI - Overexpression of the heat-shock protein 70 is associated to imatinib resistance in chronic myeloid leukemia. AB - Imatinib is an effective therapy for chronic myeloid leukemia (CML), a myeloproliferative disorder characterized by the expression of the recombinant oncoprotein Bcr-Abl. In this investigation, we studied an imatinib-resistant cell line (K562-r) generated from the K562 cell line in which none of the previously described mechanisms of resistance had been detected. A threefold increase in the expression of the heat-shock protein 70 (Hsp70) was detected in these cells. This increase was not associated to heat-shock transcription factor-1 (HSF-1) overexpression or activation. RNA silencing of Hsp70 decreased dramatically its expression (90%), and was accompanied by a 34% reduction in cell viability. Overexpression of Hsp70 in the imatinib-sensitive K562 line induced resistance to imatinib as detected by a large reduction in cell death in the presence of 1 muM of imatinib. Hsp70 level was also increased in blast cells of CML patients resistant to imatinib, whereas the level remained low in responding patients. Taken together, the results demonstrate that overexpression of Hsp70 can lead to both in vitro and in vivo resistance to imatinib in CML cells. Moreover, the overexpression of Hsp70 detected in imatinib-resistant CML patients supports this mechanism and identifies potentially a marker and a therapeutic target of CML evolution. PMID- 17109027 TI - Enhanced CD28 signaling may be a common mechanism underlying resistance to regulation. PMID- 17109028 TI - Indications for allogeneic stem cell transplantation in chronic lymphocytic leukemia: the EBMT transplant consensus. AB - The aim of this project was to identify situations where allogeneic stem cell transplantation (allo-SCT) might be considered as a preferred treatment option for patients with B-cell chronic lymphocytic leukemia (CLL). Based on a MEDLINE search and additional sources, a consented proposal was drafted, refined and approved upon final discussion by an international expert panel. Key elements of the consensus are (1) allo-SCT is a procedure with evidence-based efficacy in poor-risk CLL; (2) although definition of 'poor-risk CLL' requires further investigation, allo-SCT is a reasonable treatment option for younger patients with (i) non-response or early relapse (within 12 months) after purine analogues, (ii) relapse within 24 months after having achieved a response with purine analogue-based combination therapy or autologous transplantation, and (iii) patients with p53 abnormalities requiring treatment; and (3) optimum transplant strategies may vary according to distinct clinical situations and should be defined in prospective trials. This is the first attempt to define standard indications for allo-SCT in CLL. Nevertheless, whenever possible, allo-SCT should be performed within disease-specific prospective clinical protocols in order to continuously refine transplant indications according to new developments in risk assessment and treatment of CLL. PMID- 17109029 TI - The biosynthesis and regulation of bacterial prodiginines. AB - The red-pigmented prodiginines are bioactive secondary metabolites produced by both Gram-negative and Gram-positive bacteria. Recently, these tripyrrole molecules have received renewed attention owing to reported immunosuppressive and anticancer properties. The enzymes involved in the biosynthetic pathways for the production of two of these molecules, prodigiosin and undecylprodigiosin, are now known. However, the biochemistry of some of the reactions is still poorly understood. The physiology and regulation of prodiginine production in Serratia and Streptomyces are now well understood, although the biological role of these pigments in the producer organisms remains unclear. However, research into the biology of pigment production will stimulate interest in the bioengineering of strains to synthesize useful prodiginine derivatives. PMID- 17109030 TI - Why do we need quality-assured diagnostic tests for sexually transmitted infections? AB - The bacterial sexually transmitted infections (STIs) syphilis, gonorrhoea and chlamydia can all be cured with a single dose of antibiotic. Unfortunately, however, these infections often remain undiagnosed as many infected individuals have few if any symptoms. Diagnostic tests with high sensitivity and specificity are available for all three infections but, owing to their expense and the lack of laboratory capacity, most people in developing countries do not have access to these tests. There is a great need for simple, cheap diagnostic tests for STIs that can be performed at the point of care, enabling treatment to be given immediately. It is hoped that recent advances in our understanding of the pathogenesis of these infections, and the availability of the complete genome sequences for each causative organism, will lead to the development of improved point-of-care tests that will reduce the burden of these diseases in developing countries. PMID- 17109031 TI - Can landscape ecology untangle the complexity of antibiotic resistance? AB - Bacterial resistance to antibiotics continues to pose a serious threat to human and animal health. Given the considerable spatial and temporal heterogeneity in the distribution of resistance and the factors that affect its evolution, dissemination and persistence, we argue that antibiotic resistance must be viewed as an ecological problem. A fundamental difficulty in assessing the causal relationship between antibiotic use and resistance is the confounding influence of geography: the co-localization of resistant bacterial species with antibiotic use does not necessarily imply causation and could represent the presence of environmental conditions and factors that have independently contributed to the occurrence of resistance. Here, we show how landscape ecology, which links the biotic and abiotic factors of an ecosystem, might help to untangle the complexity of antibiotic resistance and improve the interpretation of ecological studies. PMID- 17109032 TI - The evolution and maintenance of virulence in Staphylococcus aureus: a role for host-to-host transmission? AB - Despite progress in our understanding of infectious disease biology and prevention, the conditions that select for the establishment and maintenance of microbial virulence remain enigmatic. To address this aspect of pathogen biology, we focus on two members of the Staphylococcus genus - Staphylococcus aureus and Staphylococcus epidermidis - and consider why S. aureus has evolved to become more virulent than S. epidermidis. Several hypotheses to explain this phenomenon are discussed and a mathematical model is used to argue that a complex transmission pathway is the key factor in explaining the evolution and maintenance of virulence in S. aureus. In the case of S. epidermidis, where skin contact affords easier transmission between hosts, high levels of virulence do not offer an advantage to this pathogen. PMID- 17109033 TI - Alternating divinylarene-silylene copolymers. AB - A summary of recent advances on the chemistry and photophysics of silylene-spaced divinylarene copolymers is presented. The silicon moieties have been shown to serve as an insulating spacer in these copolymers. The photophysical studies have provided useful insights into how chromophores in polymers interact intramolecularly. Because different chromophores can be regioregularly introduced into the polymeric chain, these copolymers have been extensively used as models for studying energy transfer, light harvesting as well as chiroptical transfer. PMID- 17109034 TI - Template-induced formation of heterobidentate ligands and their application in the asymmetric hydroformylation of styrene. AB - We report the template-induced formation of chelating heterobidentate ligands by the selective self-assembly of two different monodentate ligands on a rigid bis zinc(II)-salphen template with two identical binding sites; these templated heterobidentate ligands induce much higher enantioselectivities (up to 72% ee) in the rhodium-catalyzed asymmetric hydroformylation of styrene than any of the corresponding homobidentate ligands or non-templated mixed ligand combinations (up to 13% ee). PMID- 17109035 TI - Mono-palladium(II) complexes of diamidopyridine-dipyrromethane hybrid macrocycles. AB - The reaction of diamidopyridine-dipyrromethane or dipyrromethene hybrid macrocycles with palladium(II) affords mono-metalated complexes, wherein the metal centre is coordinated to the macrocycle exclusively through pyrrolic nitrogen donor atoms. PMID- 17109036 TI - Tetramethylpyridiniumporphyrazines--a new class of G-quadruplex inducing and stabilising ligands. AB - 3,4-Tetramethylpyridiniumporphyrazines bind strongly and selectively to human telomeric G-quadruplex DNA, inducing the formation of an antiparallel quadruplex in a process that mimics molecular chaperones. PMID- 17109037 TI - Free-standing nanofibrous platinum sheets and their conductivity. AB - Nanofibrous platinum sheets with a thickness of one to a few tens of nanometres were prepared over the submicron pores of polymer substrates by using long and rigid cadmium hydroxide nanostrands as templates, and these free-standing sheets gave metallic conductivity that varied greatly with the thickness. PMID- 17109038 TI - Self-assembled hexanuclear arene ruthenium metallo-prisms with unexpected double helical chirality. AB - Self-assembly of 2,4,6-tripyridyl-1,3,5-triazine (tpt) subunits with arene ruthenium building blocks and oxalato bridges affords cationic triangular metallo prisms of the type [Ru6(arene)6(tpt)2(C2O4)3]6+ (arene = C6Me6 and p Pr(i)C6H4Me); the unexpected double helical chirality of the metallo-prisms observed in the solid state persists in solution giving rise to two different stereodynamic processes as demonstrated by NMR enantiodifferentiation experiments. PMID- 17109039 TI - Signal amplification and transduction by photo-activated catalysis. AB - A simple flavin-based catalytic system is able to transform light into chemical output with amplified response utilizing a Cu(I)-catalyzed cycloaddition reaction. PMID- 17109040 TI - Fluorescently tagged polymer bioconjugates from protein derived macroinitiators. AB - BSA and lysozyme have been transformed into macroinitiators for living radical polymerisation and used to produce well-defined bioconjugates which can be fluorescently labelled providing a versatile strategy for the preparation of bioconjugates which is complementary to traditional PEGylation. PMID- 17109041 TI - A series of luminescent lanthanide-cadmium-organic frameworks with helical channels and tubes. AB - Lanthanide (Ln) oxides and cadmium (Cd) salts as sources of metals provided the first series of luminescent Ln-Cd-organic frameworks, [LnCd(imdc)(SO4)(H2O)3].0.5H2O (Ln = Tb, Eu, Dy, Gd, Er, Yb, Y, Nd, Pr; H3imdc = 4,5-imidazoledicarboxylic acid), in which the Ln atoms are linked by imdc ligands with skew coordination orientation, resulting in novel hetero-metallic-organic frameworks with left-/right-handed helical tubes (L1/R1) and channels (L2/R2) along the b axis. PMID- 17109042 TI - Spiropyran-based liquid crystals: the formation of columnar phases via acid induced spiro-merocyanine isomerisation. AB - Hexagonal columnar liquid-crystalline phases are induced for a new fan-shaped spiropyran compound as the result of an acidichromism effect of spiro-merocyanine isomerisation through protonation upon incorporation of 4-methylbenzenesulfonic acid. PMID- 17109043 TI - Single-molecule electrical studies on a 7 nm long molecular wire. AB - A self-assembled arylene-ethynylene molecular wire with a rigid 7 nm long backbone exhibits symmetrical current-voltage (I-V) characteristics and a single molecule current of 0.35 +/- 0.05 nA at 0.3 V; these data are supported by theoretical calculations. PMID- 17109044 TI - A novel soft hydrothermal (SHY) route to crystalline PbS and CdS nanoparticles exhibiting diverse morphologies. AB - We report a simple and rapid aqueous route to crystalline nanoparticles of PbS and CdS using single-source precursors and a conventional household pressure cooker. PMID- 17109045 TI - Measuring multiple carbon-nitrogen distances in natural abundant solids using R RESPDOR NMR. AB - Multiple carbon-nitrogen distances can be measured in natural abundant solids using Rotary Resonance Echo Saturation Pulse Double Resonance (R-RESPDOR) solid state NMR experiment. PMID- 17109046 TI - Entrapment of a dirhodium tetracarboxylate unit inside the aromatic bowl of a calix[4]arene: unique catalysts for C-H amination. AB - Unique calix[4]arene-derived, tetracarboxylate dirhodium(II) inclusion complexes have been prepared and evaluated as catalysts for C-H amination. PMID- 17109047 TI - Magnetically separable phase-transfer catalysts. AB - Magnetic nanoparticles-supported quaternary ammonium and phosphonium salts were prepared and evaluated as phase-transfer catalysts. Some of them exhibited good activities that were comparable to that of tetra-n-butylammonium iodide. The catalysts were readily separated using an external magnet and reusable without significant loss of their catalytic efficiency. PMID- 17109048 TI - A novel one-pot reaction involving organocopper-mediated reduction/transmetalation/asymmetric alkylation, leading to the diastereoselective synthesis of functionalized (Z)-fluoroalkene dipeptide isosteres. AB - By a novel one-pot reaction sequence involving consecutive organocopper-mediated reduction, transmetalation and asymmetric alkylation, a highly diastereoselective synthesis of functionalized (Z)-fluoroalkene dipeptide isosteres was achieved in good to excellent yields. PMID- 17109049 TI - Protein micropatterning based on electrochemically switched immobilization of bioligand on electropolymerized film of a dually electroactive monomer. AB - We demonstrate a protein micropatterning method based on electropolymerization of a monomer with two electroactive units, hydroquinone monoester and disulfide, which enables electrochemical ON-OFF switching for immobilization of bioligands on electrodes modified with the electropolymerized film. PMID- 17109050 TI - Synthesis of Cp*CH2PPh2 and its use as a ligand for the nickel-catalysed cross coupling reaction of alkyl halides with aryl Grignard reagents. AB - A new ligand, Cp*CH2PPh2 (Cp* = 1,2,3,4,5-pentamethyl-2,4-cyclopentadienyl), was prepared, and was used as a ligand for nickel-catalysed cross-coupling reaction of alkyl halides with aryl Grignard reagents, which nickel-phosphine complexes had never made possible. PMID- 17109051 TI - Rate acceleration of organic reaction by immediate solvent evaporation. AB - Several types of organic reactions were accelerated by immediate evaporation of solvents because of remarkable enhancement of molecule-to-molecule contacts between reactants. PMID- 17109052 TI - Synthesis of the bis-potassium salts of 5-hydroxy-3-oxopent-4-enoic acids and their use for the efficient preparation of 4-hydroxy-2H-pyran-2-ones and other heterocycles. AB - 5-Hydroxy-3-oxopent-4-enoic acid esters can be efficiently transformed into the stable bis-potassium salts of the corresponding 5-hydroxy-3-oxopent-4-enoic acids, from which the sensitive acids are released in situ, the latter being converted into substituted 4-hydroxy-2H-pyran-2-ones, pyrazoles and isoxazoles under mild conditions; the efficiency of this method is demonstrated by the first synthesis of two naturally occurring pyrones. PMID- 17109053 TI - Eta1 and eta2 complexes of lambda3-1,2,4,6-thiatriazinyls with CpCr(CO)x. AB - The title heterocyclic radicals coordinate to either 17e CpCr(CO)3 or 15e CpCr(CO)2 moieties as one-electron or as three-electron donors, respectively; in the former the bonding is via the perpendicular p orbital of the sulfur atom, while in the latter bonding is via p(pi) orbitals on both sulfur and nitrogen. PMID- 17109054 TI - MoS2 hierarchical hollow cubic cages assembled by bilayers: one-step synthesis and their electrochemical hydrogen storage properties. AB - Micrometer scaled MoS2 hierarchical hollow cubic cages assembled by bilayers can be synthesized via a one-step self-assembly coupled with intermediate crystal templating process without any surfactant, in which the intermediate K2NaMoO3F3 crystal formed in-situ and then served as the self-sacrificed template based on the Kirkendall Effect; The MoS2 hierarchical hollow cubic cages were employed for electrochemical hydrogen storage with a high capacity of 375 mAh g(-1) due to the more active edges exposing on the upright-standing nanoplates. PMID- 17109055 TI - s-Triazine and tri-s-triazine based organic-inorganic hybrid gels prepared from chlorosilanes by exchange reactions. AB - Hybrid polymers [(DeltaO3)4Si3]n and [(DeltaO3)SiMe]n (where Delta = C6N7 or C3N3) have been prepared by a novel sol-gel process based on exchange reactions of MeSiCl3 or SiCl4 with C6N7(OSiMe3)3 and C3N3(OSiMe3)3. PMID- 17109056 TI - Synthesis of chromophores combining second harmonic generation and two photon induced fluorescence properties. AB - The design of new chromophores presenting simultaneous SHG and TPEF properties is reported. PMID- 17109057 TI - First enantioselective organocatalytic allylation of simple aldimines with allyltrichlorosilane. AB - A new organocatalytic system, novel chiral bisformamide and in situ generated L proline-derived allylsilane reagent , which converts different aldimines to homoallylic amines in good to high yields (up to 95%) and good enantioselectivities (up to 85% ee) has been described. PMID- 17109058 TI - Structural and functional features of fructansucrases present in Leuconostoc mesenteroides ATCC 8293. AB - Glycosyltransferases produced by Leuconostoc mesenteroides subsp. mesenteroides ATCC 8293 (equivalent to NRRL B-1118) were identified. Two glucansucrases and one fructansucrases were observed in batch culture while levC and levL genes, corresponding to two fructansucrases, were isolated from information obtained from the released draft sequence of this Leuconostoc strain genome and cloned in Escherichia coli. The recombinant enzymes were shown to be fructansucrases producing a polymer identified by NMR as levan, confirming our recent report stating that these are also mosaic levansucrases bearing structural features of glucansucrases in the amino and carboxy terminal regions, as is also the case of inulosucrase (IslA) from Leuconostoc citreum CW28 and levansucrase (LevS) from L. mesenteroides NRRL B-512F. The recombinant levansucrase LevC was purified and characterized in terms of pH, temperature, and kinetic properties. The enzyme exhibits Michaelis-Menten kinetic properties with a K(m) = 27.3 mM and a k(cat) = 282.9 s(-1). This levansucrase behaves mainly as a transferase as only 30% of the substrate is hydrolyzed in a wide range of sucrose concentrations, with higher hydrolytic activities at low substrate concentrations. With this report we experimentally confirm the unusual structural pattern displayed by fructansucrases present in Leuconostoc species that group as a novel sub family of fructansucrases. PMID- 17109059 TI - Enzymatic and genetic profiles in environmental strains grown on polycyclic aromatic hydrocarbons. AB - The possible generation of oxidative stress induced by aromatic hydrocarbon degradation suggests that ancillary enzyme activities could facilitate the utilization of polycyclic aromatic hydrocarbons as sole carbon source. To investigate the metabolic profiles of low molecular weight polycyclic aromatic hydrocarbon-degrading strains of Sphingobium chlorophenolicum, Rhodococcus aetherovorans, Rhodococcus opacus and Mycobacterium smegmatis, the determination of the activity of putative detoxifying enzymes (rhodanese-like and glutathione S transferase proteins) was combined with genetic analyses. All the studied strains were able to utilize phenanthrene or naphthalene. Glutathione S-transferase activity was found in S. chlorophenolicum strains grown on phenanthrene and it was related to the presence of the bphK gene, since modulation of glutathione S transferase activity by phenanthrene paralleled the induction of glutathione S transferase transcript in the S. chlorophenolicum strains. No glutathione S transferase activity was detectable in R. aetherovorans, R. opacus and in M. smegmatis strains. All strains showed 3-mercaptopyruvate:cyanide sulfurtransferase activity. A rhodanese-like SseA protein was immunodetected in R. aetherovorans, R. opacus and in M. smegmatis strains, where increase of 3 mercaptopyruvate:cyanide sulfurtransferase activity was significantly induced by growth on phenanthrene. PMID- 17109060 TI - Prognostic factors of radiographic progression in early rheumatoid arthritis: a two year prospective study after a structured therapeutic strategy using DMARDs and very low doses of glucocorticoids. AB - The objective of the study was to analyze the prognostic factors of radiographic progression in a series of patients with early rheumatoid arthritis (RA) after 2 years of therapy with a structured algorithm using disease-modifying antirheumatic drugs (DMARDs) and very low doses of oral glucocorticoids. One hundred and five patients (81% female) with early RA (disease duration <2 years) treated with the same therapeutic protocol using gold salts and methotrexate in a step-up strategy, together with methylprednisolone (4 mg/day), were followed up for 2 years. The outcome variable was radiographic progression after 2 years of DMARD therapy using the modified Larsen method. Clinical, biological, immunogenetic, and radiographic data were analyzed at study entry and after 1 and 2 years of follow-up. Radiographic progression (increase of four or more units in the Larsen score) was observed in 32% of patients after 2 years of follow-up. The percentage of erosive disease increased from 18.3% at baseline to 28.9% at 12 months and 44.6% at 24 months, in spite of a significant improvement in disease activity. New erosions appeared in 33% of patients after 2 years. Several baseline parameters were associated with radiographic progression in the univariate analysis: shared epitope (SE) homozygozity, HLA-DRB*04 alleles, female gender, hemoglobin, erythrocyte sedimentation rate, and anticyclic citrullinated peptide antibodies (anti-CCP). In the multivariate analysis, female gender [odds ratio (OR) 5.5, 95% confidence interval (CI): 1.1-28.2, p = 0.04], DRB1*04 alleles (OR 3.1, 95% CI 1.1-9, p = 0.03) and, marginally, anti-CCP antibodies (OR 3.6, 95% CI 0.9-14.5, p = 0.06), were associated with progression. Female patients with both DRB1*04 alleles and anti-CCP antibodies showed the highest scores in radiographic progression. The presence, but not the titer, of anti-CCP antibodies predicted progression. The positive predictive value of the multivariate model for progression was only 53.9% whereas the negative predictive value was 80.3%. In a series of early RA patients treated with a structured algorithm using DMARDs and very low doses of glucocorticoids, radiographic progression was observed in one third of patients after 2 years. Female gender, DRB1*04 alleles (rather than the SE), and the presence of anti-CCP antibodies at baseline (independently of the titer) were the most important predictors of progression. The utility of these parameters in clinical practice is limited by their relatively low positive predictive value. PMID- 17109061 TI - Links between cardiovascular disease and osteoporosis in postmenopausal women: serum lipids or atherosclerosis per se? AB - INTRODUCTION AND HYPOTHESIS: Epidemiological observations suggest links between osteoporosis and risk of acute cardiovascular events and vice versa. Whether the two clinical conditions are linked by common pathogenic factors or atherosclerosis per se remains incompletely understood. We investigated whether serum lipids and polymorphism in the ApoE gene modifying serum lipids could be a biological linkage. METHODS: This was an observational study including 1176 elderly women 60-85 years old. Women were genotyped for epsilon (epsilon) allelic variants of the ApoE gene, and data concerning serum lipids (total cholesterol, triglycerides, HDL-C, LDL-C, apoA1, ApoB, Lp(a)), hip and spine BMD, aorta calcification (AC), radiographic vertebral fracture and self-reported wrist and hip fractures, cardiovascular events together with a wide array of demographic and lifestyle characteristics were collected. RESULTS: Presence of the ApoE epsilon 4 allele had a significant impact on serum lipid profile, yet no association with spine/hip BMD or AC could be established. In multiple regression models, apoA1 was a significant independent contributor to the variation in AC. However, none of the lipid components were independent contributors to the variation in spine or hip BMD. When comparing the women with or without vertebral fractures, serum triglycerides showed significant differences. This finding was however not applicable to hip or wrist fractures. After adjustment for age, severe AC score (>or=6) and/or manifest cardiovascular disease increased the risk of hip but not vertebral or wrist fractures. CONCLUSION: The contribution of serum lipids to the modulators of BMD does not seem to be direct but rather indirect via promotion of atherosclerosis, which in turn can affect bone metabolism locally, especially when skeletal sites supplied by end-arteries are concerned. Further studies are needed to explore the genetic or environmental risk factors underlying the association of low triglyceride levels to vertebral fractures. PMID- 17109062 TI - The prevalence of vertebral fracture amongst patients presenting with non vertebral fractures. AB - INTRODUCTION: Despite vertebral fracture being a significant risk factor for further fracture, vertebral fractures are often unrecognised. A study was therefore conducted to determine the proportion of patients presenting with a non vertebral fracture who also have an unrecognised vertebral fracture. METHODS: Prospective study of patients presenting with a non-vertebral fracture in South Glasgow who underwent DXA evaluation with vertebral morphometry (MXA) from DV5/6 to LV4/5. Vertebral deformities (consistent with fracture) were identified by direct visualisation using the Genant semi-quantitative grading scale. RESULTS: Data were available for 337 patients presenting with low trauma non-vertebral fracture; 261 were female. Of all patients, 10.4% were aged 50-64 years, 53.2% were aged 65-74 years and 36.2% were aged 75 years or over. According to WHO definitions, 35.0% of patients had normal lumbar spine BMD (T-score -1 or above), 37.4% were osteopenic (T-score -1.1 to -2.4) and 27.6% osteoporotic (T-score -2.5 or lower). Humerus (n=103, 31%), radius-ulna (n=90, 27%) and hand/foot (n=53, 16%) were the most common fractures. For 72% of patients (n=241) the presenting fracture was the first low trauma fracture to come to clinical attention. The overall prevalence of vertebral deformity established by MXA was 25% (n=83); 45% (n=37) of patients with vertebral deformity had deformities of more than one vertebra. Of the patients with vertebral deformity and readable scans for grading, 72.5% (58/80) had deformities of grade 2 or 3. Patients presenting with hip fracture, or spine T-score 128 microg/ml, >128 microg/ml, 0.25 microg/mL, 1 microg/ml, 0.12 microg/ml, and 0.5 g/ml, respectively. It is suggest that the minimum bactricidal concentration (MBC) was dissociated from the minimum inhibitory concentration (MIC) in S. pneumoniae and H. influenzae with abnormal pbp genes. PMID- 17109100 TI - Heart rate variability reveals risk of arrhythmias after intoxication with antidepressants. PMID- 17109101 TI - [Molecular targeted therapy]. AB - Until recently, cancer therapy was based on three modalities: surgery, radiotherapy, and cytostatic chemotherapy. In most instances treatment of solid tumors was a surgical domain. For patients with incomplete resection or relapse after surgery, radiotherapy and chemotherapy usually offered only partial response and mostly of limited duration. By the mid-1990s visions of antibody based therapies, vaccination strategies, and even gene-specific therapies existed but seemed far from clinical practice. United States Federal Drug Administration approval of the humanized antibody rituximab (1997) and the tyrosine kinase inhibitor imatinib (2001) has changed perceptions of oncologic treatment. These drugs turned visions into reality and led the pharmaceutical industry, clinicians, and patients to new perspectives. This article gives an overview of the development of this fourth modality in cancer therapy, so-called targeted therapy. PMID- 17109102 TI - Adhesion molecules in chronic ulcerative colitis. AB - BACKGROUND AND AIMS: The adhesion molecule expression in colonic mucosa is pivotal for transition from quiescent to active stage of ulcerative colitis (UC). The aim of the present study is to reveal the adhesion molecule profile of colonic mucosa in the active stage of UC and in remission. MATERIALS AND METHODS: Biopsy specimens obtained from 14 patients with UC (seven with active disease and seven with UC in remission) and from seven controls were used. Immunohistochemistry was performed with antibodies against ICAM-1, VCAM-1, E selectin, LFA-1, Mac-1, and VLA-4. RESULTS: In controls, slight ICAM-1 positivity was observed on thety endothelium of blood vessels of the mucosal and submucosal layer and only single ICAM-1-, Mac-1-, and LFA-1-positive cells were found. In all patients with UC, the endothelium of venules in the edematous mucosal and submucosal layers was ICAM-1-, VCAM-1-, and E-selectin-positive. Numerous ICAM-1- and LFA-1-positive and less VCAM-1-, Mac-1-, and VLA-4-positive inflammatory cells were detected in mucous layers of acute UC. In specimens of UC in remission, the inflammatory cells positive for the studied adhesion molecules were significantly less in number in the mucosa and submucosa (p < 0.05). CONCLUSIONS: Based on the increased expression of ICAM-1, VCAM-1, and their ligands LFA-1 and VLA-4 in patients with UC, we can conclude that these adhesion molecules play a key role in the adherence of lymphocytes and macrophages to endothelial cells maintaining the chronic inflammation. Presence of E-selectin on endothelial cells of venules could be a sign of relapse after remission in UC. PMID- 17109103 TI - The role of chemotherapy in microsatellite unstable (MSI-H) colorectal cancer. AB - INTRODUCTION: High-frequency microsatellite instability (MSI-H) is an alternate pathway of colorectal carcinogenesis, which accounts for 15% of all sporadic colorectal cancers. These tumours arise from mutations in the DNA mismatch repair system and thus have different responses to chemotherapeutic agents compared to microsatellite stable (MSS) cancers. OBJECTIVE: This review aims to summarise the available literature on the responses to chemotherapy in MSI-H colorectal cancer (CRC). RESULTS AND DISCUSSION: 5 Fluorouracil (5FU) is commonly used as a chemotherapeutic agent in colon cancer and in vitro evidence shows reduced response to 5FU in MSI-H CRC. The clinical evidence is conflicting but favours a reduced response to 5FU in MSI-H CRC. Several newer agents such as COX-2 inhibitors and irinotecan are also reviewed. CONCLUSION: Available evidence suggests that MSI-H CRC have different behaviour patterns and response to chemotherapy compared with MSS CRC. PMID- 17109104 TI - Matrix metalloproteinases in rectal mucosa, tumour and plasma: response after preoperative irradiation. AB - BACKGROUND: In rectal cancer treatment, preoperative radiotherapy has led to reduction of local recurrence, but it is associated with morbidity and increased risk for secondary tumours. Matrix metalloproteinases (MMPs) are associated with tumour progression through tissue remodeling. The aim of this study was to investigate tissue remodeling after preoperative radiotherapy and to explore possible correlations with clinical outcome. MATERIALS AND METHODS: Ninety-one patients scheduled for rectal cancer surgery were included; 49% received preoperative radiotherapy three-field treatment, 5 x 5 Gy. Blood samples and biopsies from tumour and adjacent mucosa were taken during surgery. Biopsies and plasma were assayed with ELISA for MMP-1, MMP-2 and MMP-9. Clinical outcome was reviewed focusing on infections, perineal healing, fistula formation, anastomotic dehiscence, small bowel obstruction, local recurrence and distant metastases. RESULTS: Compared to non-irradiated mucosa, MMP-2 (p < 0.0001), MMP-1 (p = 0.03) and MMP-9 (p = 0.04) were significantly higher in irradiated normal mucosa. Tumour tissue had higher levels of MMP-2 if irradiated (p < 0.0001). A correlation between MMP-2 levels and wound infection (p = 0.02) as well as fistula formation (p = 0.04) was found. MMP-1 in mucosa (p = 0.02) and tumour (p = 0.04) were higher in patients developing distant metastases. Plasma levels were not influenced by irradiation, but MMP-2 was higher in patients who were later developing distant metastases (p = 0.007). CONCLUSIONS: Extracellular matrix remodeling after radiotherapy seems to be correlated to postoperative morbidity; MMP-2 is associated with both wound infections and fistula formation. High levels of MMP-1 in tumour and mucosa as well as MMP-2 in plasma may be correlated to risk of developing distant metastases. PMID- 17109105 TI - Pulmonary resection for metastases from colorectal cancer: prognostic factors and survival. AB - BACKGROUND: Pulmonary metastases occur in up to 10% of all patients who undergo curative resection. Surgical resection is an important part in the treatment of pulmonary metastasis from colorectal cancer. We analyzed the treatment outcome and prognostic factors affecting survival in this subset of patients. MATERIALS AND METHODS: Between October 1994 and December 2004, 59 patients underwent curative resection for pulmonary metastases of colorectal cancer. Uncontrollable synchronous liver and lung metastasis or synchronous colorectal cancer with isolated lung metastasis were excluded from this study. A retrospective review of patient characteristics and factors influencing survival was performed. Survival was analyzed by the Kaplan-Meier method. Comparison between groups were performed by a log-rank analysis and the Cox proportional hazard model. RESULTS: The 5-year overall survival rate of all patients who received pulmonary resection was 50.3%. The number of pulmonary metastases was significantly related with survival in univariate analysis, but not in multivariate analysis (p = 0.032). Prethoracotomy carcinoembryonic antigen (CEA) level exceeding 5 ng/ml was related with poor survival (p = 0.001). A disease-free interval of greater than 2 years did not correlate with survival after thoracotomy (p = 0.3). CONCLUSION: The prethoracotomy CEA level and the number of metastases were independent prognostic factors. Resection of pulmonary metastasis from colorectal cancer may result in improved survival or even healing in selected patients. Pulmonary resection of colorectal cancer is regarded as a safe and effective treatment with low morbidity and mortality rates. PMID- 17109106 TI - Timing of thoracic and lumbar fracture fixation in spinal injuries: a systematic review of neurological and clinical outcome. AB - A systematic review of all available evidence on the timing of surgical fixation for thoracic and lumbar fractures with respect to clinical and neurological outcome was designed. The purpose of this review is to clarify some of the controversy about the timing of surgical fracture fixation in spinal trauma. Better neurological outcome, shorter hospital stay and fewer complications have been reported after early fracture fixation. But there are also studies showing no difference in neurological outcome when compared to late treatment. Mortality is another controversial point since a recent report of higher mortality in early treated patients. A systematic review of the literature was preformed. Ten articles were included. Early fracture fixation is associated with less complications, shorter hospital and ICU stay. The effect of early treatment on the neurological outcome remains unclear due to the contradictory results of the included studies. Early thoracic and lumbar fracture fixation results in improvement of clinical outcome, but the effect on neurological outcome remains controversial. PMID- 17109107 TI - Effects of acetylcholine on coding of taste information in the primary gustatory cortex in rats. AB - Acetylcholine (ACh) receptors are widely distributed throughout the cerebral cortex in rats. Recently, cholinergic innervation of the gustatory cortex (GC) was reported to be involved in certain taste learning in rats. Here, the effects of iontophoretic application of ACh on the response properties of GC neurons were studied in urethane-anesthetized rats. ACh affected spontaneous discharges in a small fraction of taste neurons (11 of 86 neurons tested), but influenced taste responses in 27 of 43 neurons tested. No correlations with ACh susceptibility were noted for spontaneous discharges and taste responses. Among the 27 neurons, ACh facilitated taste responses in 13, inhibited taste responses in 13 and either facilitated or inhibited taste responses depending on the stimuli in 1. Furthermore, ACh affected the responses to best stimuli that produced the largest responses among four basic tastants (best responses) in 7 of 27 taste neurons, to non-best responses in 9, and to both best and non-best responses in 11. ACh mostly inhibited the best responses (13 of 18 neurons). Thus, ACh often decreased the response selectivity to the four basic tastants and changed the response profile. Atropine, a general antagonist of muscarinic receptors, antagonized ACh actions on taste responses or displayed the opposite effects on taste responses to ACh actions in two-thirds of the neurons tested. These findings indicate that ACh mostly modulates taste responses through muscarinic receptors, and suggest that ACh shifts the state of the neuron network in the GC, in terms of the response selectivities and response profiles. PMID- 17109108 TI - Superadditive BOLD activation in superior temporal sulcus with threshold non speech objects. AB - Evidence from neurophysiological studies has shown the superior temporal sulcus (STS) to be a site of audio-visual integration, with neuronal response to audio visual stimuli exceeding the sum of independent responses to unisensory audio and visual stimuli. However, experimenters have yet to elicit superadditive (AV > A+V) blood oxygen-level dependent (BOLD) activation from STS in humans using non speech objects. Other studies have found integration in the BOLD signal with objects, but only using less stringent criteria to define integration. Using video clips and sounds of hand held tools presented at psychophysical threshold, we were able to elicit BOLD activation to audio-visual objects that surpassed the sum of the BOLD activations to audio and visual stimuli presented independently. Our findings suggest that the properties of the BOLD signal do not limit our ability to detect and define sites of integration using stringent criteria. PMID- 17109109 TI - Visual information throughout a reach determines endpoint precision. AB - People make rapid, goal-directed movements to interact with their environment. Because these movements have consequences, it is important to be able to control them with a high level of precision and accuracy. Our hypothesis is that vision guides rapid hand movements, thereby enhancing their accuracy and precision. To test this idea, we asked observers to point to a briefly presented target (110 ms). We measured the impact of visual information on endpoint precision by using a shutter to close off view of the hand 50, 110 and 250 ms into the reach. We found that precision was degraded if the view of the hand was restricted at any time during the reach, despite the fact that the target disappeared long before the reach was completed. We therefore conclude that vision keeps the hand on the planned trajectory. We then investigated the effects of a perturbation of target position during the reach. For these experiments, the target remained visible until the reach was completed. The target position was shifted at 110, 180 or 250 ms into the reach. Early shifts in target position were easily compensated for, but late shifts led to a shift in the mean position of the endpoints; observers pointed to the center of the two locations, as a kind of best bet on the position of the target. Visual information is used to guide the hand throughout a reach and has a significant impact on endpoint precision. PMID- 17109110 TI - Modulation of anticipatory postural adjustments associated with unloading perturbation: effect of characteristics of a motor action. AB - The purpose of the study was to determine whether characteristics of a motor action affect anticipatory postural adjustments (APAs). Standing subjects held a load between their hands with arms extended in front of their body. Next, subjects performed bilateral shoulder abduction movements (motor action) of three amplitudes at three instructed speeds. This motor action led to the release of the same load, inducing unloading perturbation in the sagittal plane. Electromyographic activities were recorded for the leg and trunk muscles. A change in the background muscle activity in these muscles was observed prior to the unloading perturbation and was quantified as APAs. APAs were dependant on instructed speed of the motor action; larger APA activities were observed in the leg and trunk muscles with a faster speed instruction. Meanwhile, the modulation of APAs was not observed by altering the movement amplitude. Moreover, experiments showed that motor action itself without a load release did not generate APA activity. Therefore, we concluded that the central nervous system selects information within a motor action (i.e., speed instruction) to approximate the magnitude of the forthcoming perturbation and modulate APAs, even when the unloading perturbation was unchanged. PMID- 17109111 TI - Bilateral motor unit synchronization is functionally organized. AB - To elucidate the neural interactions underlying bimanual coordination, we investigated in 11 participants the bilateral coupling of homologous muscles in an isometric force production task involving fatiguing elbow flexion and extension. We focused on changes in motor unit (MU) synchronization as evident in EMG recordings of relevant muscles. In contrast to a related study on leg muscles, the arm muscles did not exhibit MU synchronization around 16 Hz, consistent with our hypothesis that 16 Hz MU synchronization is linked to balance maintenance. As expected, bilateral MU synchronization was apparent between 8 and 12 Hz and increased with fatigue and more strongly so for extensor than for flexor muscles. MU synchronization in that frequency band is interpreted in terms of common bilateral input and substantiates the idea that common input is functionally organized. Since these findings are consistent with the literature on mirror movements, they suggest that both phenomena may be related. PMID- 17109112 TI - Metastatic melanoma in a pediatric patient. PMID- 17109113 TI - [Importance of intrathecal pain therapy]. AB - Intraspinal drug infusion using implantable pumps and catheter systems is a safe and effective therapy for selected pain patients with severe chronic pain. It improves pain relief, reduces drug-related side effects, decreases the need for oral analgesia and enhances quality of life in a segment of chronic pain patients whose pain has not been controlled with more conservative therapies. Intrathecal drug therapy has therefore established its role in the treatment of malignant pain, benign pain and severe spasticity.Careful patient selection and management as well as a multidisciplinary approach are determinants of successful treatment. Current practices for patient selection and management, screening, drug selection, dosing and implantation for intrathecal drug delivery systems are discussed. PMID- 17109114 TI - [Influence of negative coping style on post-operative pain reporting and pain related behaviour]. AB - OBJECTIVE: The aim of the study is to show whether negative coping style has an impact on post-operative pain reporting and pain-related behaviour. METHODS: After informed consent 48 gynaecological patients were enrolled in the study. Negative coping style was assessed by using the stress coping questionnaire SVF 78 (Janke and Erdmann). Patients were dichotomised by scores of the negative coping style. Pain reporting concerned pain intensity and pain quality. Pain related behaviour was assessed by patient-controlled analgesia (PCA). Pain intensity and pain quality were assessed prior to surgery and 24 h after surgery. Patient-controlled analgesia was recorded up to 24 h after surgery. Trait anxiety (STAI-X2) was used as a control variable. RESULTS: Patients with high scores in negative coping style report increased levels of pain in both affective and sensory pain perception. At the same time patient-controlled analgesia is used less frequently by this group. This result remains significant considering trait anxiety as a covariate. CONCLUSIONS: High negative coping style may cause a discrepancy between pain reporting and pain-related behaviour. This should be taken into account in the perioperative care of patients. PMID- 17109115 TI - [Symptom checklists in patients with chronic pain]. AB - Symptom checklists, in particular the SCL-90-R, are widely used in screening for psychological distress in patients with chronic pain. A detailed analysis reveals that this may lead to wrong diagnoses in many cases. The problem of using the SCL 90-R in patients with chronic pain is (a) that scale means are elevated due to items asking for pain-associated symptoms and (b) that there is a lack of specificity of the various dimensions of the SCL-90-R. Numerous short forms of the SCL-90-R are compared to the original one and analysed with respect to their applicability in patients with chronic pain. Other screening questionnaires not related to the SCL-90-R were analysed for the same purpose. As a result, the SCL 27, K9, HADS and PHQ are judged to be applicable for screening in patients with chronic pain. PMID- 17109116 TI - [The influence of somatic dysfunction on chronic muscular skeletal pain syndromes]. AB - Chronic muscular skeletal pain syndromes (CMSPS) are one of the major health issues. Despite progress in research, treatment and diagnosis remain difficult. The aim of this study was to examine the influence of somatic dysfunction on CMSPS. A total of 216 patients were examined in a standardized way to assess the influence of morphological and psychosocial factors as well as of somatic dysfunction on CMSPS. Measurements were taken at admission. The results showed a statistically significant link between somatic dysfunction, pain chronicity, pain distribution, and pain-related disability. Since there was no or only a minor correlation between somatic dysfunction and the results of the psychometric testing and the morphological findings, somatic dysfunction should be regarded as an independent factor influencing CMSPS and be further scientifically evaluated. PMID- 17109117 TI - Induced secretion of insulin-like growth factor binding protein-1 (IGFBP-1) in human hepatoma cell HepG2 by rubratoxin B. AB - The induction of insulin-like growth factor binding protein-1 (IGFBP-1) secretion by rubratoxin B was investigated using human hepatoma cell line HepG2; we also documented the involvement of stress-activated MAP kinases [c-Jun-N-terminal kinases (JNKs) and p38s] in this process. Rubratoxin B dramatically enhanced IGFBP-1 secretion, which peaked at a concentration of 40 microg/ml. The amount of IGFBP-1 mRNA increased with time and plateaued at 6 h. Compared with the amounts of IGFBP-1 secreted, the induction ratios of transcription were much smaller, indicating that IGFBP-1 secretion is regulated chiefly post-transcriptionally. The result of concomitant treatment with rubratoxin B and JNK inhibitor indicated that JNKs do not affect rubratoxin B-induced IGFBP-1 secretion. Alternatively, rubratoxin B-associated induction of IGFBP-1 secretion was marked in the absence of p38 inhibitor but attenuated in its presence. Therefore, p38s appear to stimulate rubratoxin B-induced IGFBP-1 secretion. Treatment with p38 inhibitor slightly increased the amount of rubratoxin B-induced IGFBP-1 mRNA. However this induction ratio was smaller than that of rubratoxin B-induced secretion, suggesting that p38s regulate IGFBP-1 secretion both transcriptionally and post transcriptionally. In this study, we showed that rubratoxin B induces IGFBP-1 levels in HepG2 cells and p38s contribute to this process. PMID- 17109118 TI - Prominin-2 is a cholesterol-binding protein associated with apical and basolateral plasmalemmal protrusions in polarized epithelial cells and released into urine. AB - Prominin-2 is a pentaspan membrane glycoprotein structurally related to the cholesterol-binding protein prominin-1, which is expressed in epithelial and non epithelial cells. Although prominin-1 expression is widespread throughout the organism, the loss of its function solely causes retinal degeneration. The finding that prominin-2 appears to be restricted to epithelial cells, such as those found in kidney tubules, raises the possibility that prominin-2 functionally substitutes prominin-1 in tissues other than the retina and provokes a search for a definition of its morphological and biochemical characteristics. Here, we have investigated, by using MDCK cells as an epithelial cell model, whether prominin-2 shares the biochemical and morphological properties of prominin-1. Interestingly, we have found that, whereas prominin-2 is not restricted to the apical domain like prominin-1 but is distributed in a non polarized fashion between the apical and basolateral plasma membranes, it retains the main feature of prominin-1, i.e. its selective concentration in plasmalemmal protrusions; prominin-2 is confined to microvilli, cilia and other acetylated tubulin-positive protruding structures. Similar to prominin-1, prominin-2 is partly associated with detergent-resistant membranes in a cholesterol-dependent manner, suggesting its incorporation into membrane microdomains, and binds directly to plasma membrane cholesterol. Finally, prominin-2 is also associated with small membrane particles that are released into the culture media and found in a physiological fluid, i.e. urine. Together, these data show that all the characteristics of prominin-1 are shared by prominin-2, which is in agreement with a possible redundancy in their role as potential organizers of plasma membrane protrusions. PMID- 17109119 TI - Adaptive changes in structure of skeletal muscles from adult Sod1 homozygous knockout mice. AB - Cu/Zn superoxide dismutase (SOD1), which is localized cytoplasmically and in the mitochondrial intermembrane space, is an enzyme that is critically important for superoxide free-radical elimination. Compared with age-matched wild-type littermates (Sod1 ( +/+ )), SOD1 homozygous knockout (Sod1 ( -/- )) mice have smaller body masses, heart and skeletal muscle masses, and muscle cross-sectional areas. At the light-microscopic level, cross sections of skeletal muscles from Sod1 ( -/- ) mice show no gross structural abnormalities. Following the staining of muscles of Sod1 ( -/- ) mice for succinate dehydrogenase (SDH) enzymatic activity, a grouping of SDH-positive fibers has been observed. Immunostaining for neural cell adhesion marker in the gastrocnemius muscle of Sod1 ( -/- ) mice has revealed a small number of atrophic denervated muscle fibers. No denervated fibers are observed in extensor digitorum longus (EDL), tibialis anterior, or plantaris muscles. An increase in mRNA expression levels of myogenin and acetylcholine receptor alpha has been detected in muscles in Sod1 ( -/- ) mice, but no changes in MyoD expression occur. Compared with fast oxidative fibers in EDL muscles of Sod1 ( +/+ ) mice, those of Sod1 ( -/- ) mice show increased accumulations of sub-sarcolemmal mitochondria. We conclude that the lack of SOD1 in adult Sod1 ( -/- ) mice does not result in extensive denervation of skeletal muscle fibers, although the distribution of fiber types is modified, and that fast oxidative fibers develop alterations in the amount and spatial distribution of sub-sarcolemmal mitochondria. PMID- 17109120 TI - Human and rodent bone marrow mesenchymal stem cells that express primitive stem cell markers can be directly enriched by using the CD49a molecule. AB - Bone marrow (BM) from human and rodent species contains a population of multipotential cells referred to as mesenchymal stem cells (MSCs). Currently, MSCs are isolated indirectly by using a culture step and then the generation of fibroblast colony-forming units (CFU-fs). Unprocessed or native BM MSCs have not yet been fully characterised. We have previously developed a direct enrichment method for the isolation of MSCs from human BM by using the CD49a protein (alpha1 integrin subunit). As the CD49a gene is highly conserved in mammals, we have evaluated whether this direct enrichment can be employed for BM cells from rodent strains (rat and mouse). We have also studied the native phenotype by using both immunodetection and immunomagnetic methods and have compared MSCs from mouse, rat and human BM. As is the case for human BM, we have demonstrated that all rodent multipotential CFU-fs are contained within the CD49a-positive cell population. However, in the mouse, the number of CFU-fs is strain-dependent. Interestingly, all rat and mouse Sca-1-positive cells are concentrated within the CD49a-positive fraction and also contain all CFU-fs. In human, the colonies have been detected in the CD49a/CD133 double-positive population. Thus, the CD49a protein is a conserved marker that permits the direct enrichment of BM MSCs from various mammalian species; these cells have been phenotyped as true BM stem cells. PMID- 17109121 TI - [Necrosis of the femoral head in late pregnancy. Necessity of early diagnosis]. AB - Limping and groin pain can issue diagnostic problems during late pregnancy. Differential diagnosis of two idiopathic syndromes, transient osteoporosis and osteonecrosis of the femoral head, is made possible by MRI in the early stages. This case is reported to demonstrate the need to distinguish between those syndromes early so as to prevent further joint damage. PMID- 17109122 TI - Identification and expression of natriuretic peptide receptor type-A and -B mRNA in freshwater and seawater rainbow trout. AB - Natriuretic peptide receptors mediate the physiological response of natriuretic peptide hormones. One of the natriuretic peptide receptor types is the particulate guanylyl cyclase receptors, of which there are two identified: NPR-A and NPR-B. In fishes, these have been sequenced and characterized in eels, medaka, and dogfish shark (NPR-B only). The euryhaline rainbow trout provides an opportunity to further pursue examination of the system in teleosts. In this study, partial rainbow trout NPR-A-like and NPR-B-like mRNA sequences were identified via PCR and cloning. The sequence information was used in real-time PCR to examine mRNA expression in a variety of tissues of freshwater rainbow trout and rainbow trout acclimated to 35 parts per thousand seawater for a period of 10 days. In the excretory kidney and posterior intestine, real-time PCR analysis showed greater expression of NPR-B in freshwater fish than in those adapted to seawater; otherwise, there was no difference in the expression of the individual receptors in fresh water or seawater. In general, the expression of the NPR-A and NPR-B type receptors was quite low. These findings indicate that NPR-A and NPR-B mRNA expression is minimally altered under the experimental regime used in this study. PMID- 17109123 TI - Study of the dynamical properties of water in disaccharide solutions. AB - This work presents quasi-elastic neutron scattering (QENS) and neutron spin echo (NSE) findings on homologous disaccharide (i.e. trehalose, maltose and sucrose)/water solutions as a function of temperature. The dynamical properties of these systems are investigated by QENS, which, on the picosecond scale, allows for the characterisation of the diffusion of both solutes and solvent. On the other hand, NSE investigates the dynamics on the nanosecond scale, allowing for the relaxation times of the disaccharide/water systems to be evaluated. The experimental data highlight a strong slowing down of water in the presence of disaccharides. The whole set of findings indicates, therefore, a noticeable disaccharide-water interaction, which is more intense in the case of trehalose. This feature can justify its higher bioprotective effectiveness. PMID- 17109124 TI - Fulminant endogenous Klebsiella pneumoniae endophthalmitis: imaging findings. AB - Endogenous endophthalmitis is a rare but serious complication of Klebsiella pneumoniae infection that has been predominantly reported to occur in diabetic patients of East Asian origin with K. pneumoniae liver abscesses. The clinical symptoms and signs of endogenous endophthalmitis are nonspecific and may mimic other causes of an acute non-traumatic eye. Permanent visual loss and blindness are common sequelae because of delayed diagnosis and treatment. Computed tomography and magnetic resonance imaging can help to differentiate endophthalmitis from other causes of acute non-traumatic eye when the clinical findings are equivocal. Fluid attenuated inversion recovery and diffusion weighted imaging with apparent diffusion coefficient map are superior to T2 weighted and gadolinium-enhanced T1-weighted sequences in demonstrating intra ocular abscesses. PMID- 17109125 TI - Gastric pneumatosis following nasogastric tube placement: a case report with literature review. AB - We report a case of gastric emphysema following placement of nasogastric (NG) tube. Gas in the wall of the stomach is a rare finding seen in various clinical situations. The reported cases fall into two different categories: "gastric emphysema" and "emphysematous gastritis". Differentiating these two entities is important, as the first one is usually a benign condition, but the second one carries a poor prognosis. PMID- 17109126 TI - Blunt traumatic injury of the ascending aorta: multidetector CT findings in two cases. AB - Blunt ascending aortic injuries are rare in clinical practice. We have encountered two types of injuries to the ascending aorta with multidetector computed tomography: (1) a tear of the wall of the aortic root with a contained rupture and associated hemopericardium and (2) a tear at the level of the aortic valve cusp without associated hemopericardium. In reviewing our experience with aortic trauma at our institution under IRB waiver of consent, we encountered two cases of ascending aortic rupture that illustrate the two injury patterns. We present these two cases to alert radiologists to the multidetector computed tomographic findings of this life-threatening injury. PMID- 17109127 TI - Handlebar hernia: a case report. AB - A 36-year-old man fell from his bicycle and struck his lower abdomen on the end of the handlebar. Computed tomography (CT) showed a small bowel loop protruding into the subcutaneous fat layer of the abdominal wall. We present this case of handlebar hernia, a rare type of traumatic abdominal wall hernia, and the usefulness of CT in diagnosing such injuries. PMID- 17109128 TI - Acute obstruction of the afferent loop caused by an enterolith. AB - Afferent loop obstruction is a relatively rare but significant complication of Billroth II gastrojejunostomy. We report the imaging findings in a patient in whom obstruction presented acutely and was due to the presence of an enterolith. CT showed dilatation of both the main pancreatic duct and the biliary ducts, and a markedly dilated afferent loop within which a 5-cm mass was present. The lesion had a heterogeneous, laminated appearance and did not show any contrast enhancement. Edema of fatty tissues surrounding the pancreatic tail, which extended to the left pararenal spaces, a small amount of free peritoneal fluid surrounding the spleen, and an aneurysm of the splenic artery of about 3 cm were also present. The diagnosis of afferent loop obstruction has to be considered in patients with previous Billroth II gastrojejunostomy who present with acute abdominal pain and laboratory findings indicating pancreatitis. Although rarely, an enterolith can be the cause of obstruction. CT allows to establish the diagnosis. PMID- 17109129 TI - The roles of endogenous reactive oxygen species and nitric oxide in triptolide induced apoptotic cell death in macrophages. AB - Triptolide, a major active component extracted from the root of Tripterygium wilfordii Hook f, has been shown to possess potent immunosuppressive and anti inflammatory properties. In the present report, we reported that triptolide increased the generation of reactive oxygen species (ROS) and nitric oxide (NO) and induced apoptosis of RAW 264.7 cells in a dose-dependent manner (5-25 ng/ml). The antioxidant, reduced glutathione (GSH), significantly inhibited triptolide induced apoptosis and inhibited the degradation of Bcl-2 protein, disruption of mitochondrial membrane potential, release of cytochrome c from mitochondria into the cytosol, activation of caspase-3, and cleavage of poly-(ADP-ribose) polymerase. The inducible nitric oxide synthase-specific inhibitor 1400w blocked triptolide-induced apoptosis, but did not alter mitochondria disruption and caspase-3 activation. These results, for the first time, implicated that the increased endogenous ROS and NO co-mediated triptolide-induced apoptosis in macrophages. ROS initiated triptolide-induced apoptosis by the mitochondria signal pathway, while the apoptotic cell death mediated by NO was not via mitochondria collapse and caspase-3 activation. In addition, combining mathematical calculation and computer simulation based on our conventional experimental results, we set and validated the apoptotic model and provided more dynamic processes of triptolide-induced apoptotic cascade in macrophages. PMID- 17109131 TI - Diversity and comparability--towards a common European Higher Education Area. PMID- 17109130 TI - CD8(+) T cells armed with retrovirally transduced IFN-gamma. AB - Interferon-gamma (IFN-gamma) is considered a key cytokine involved in the preventive and defensive responses of T cells against infectious pathogens and tumors. Therefore, the transgenic expression of IFN-gamma in specific T cells appears to be an obvious therapeutic possibility. To directly examine whether IFN gamma production can be increased in T cells, we introduced an IFN-gamma encoding cDNA into IFN-gamma(-/-) and IFN-gamma(+/+) CD8(+) effector populations by retroviral transduction. Here, we show that CD8 T cells can be equipped with IFN gamma that increases their capacity to secrete the cytokine. Despite constitutive retroviral IFN-gamma mRNA transcription, translation and secretion of IFN-gamma protein was tightly regulated and only observed in activated T cells. Neither proliferation nor cytolytic activity of CTL was affected by IFN-gamma transduction. Importantly, CD8(+) T cells retrovirally transduced with IFN-gamma exhibit augmented tumor suppressive capacity upon adoptive transfer into IFN gamma(-/-) mice. Thus, T cells can be readily armed with IFN-gamma without risking immunopathology by dysregulated production of this highly potent proinflammatory cytokine. PMID- 17109132 TI - Health care: another challenge for analytical chemistry. PMID- 17109133 TI - Advantages of downsizing electrochemical detection for DNA assays. PMID- 17109134 TI - Analytical procedure for determination of the time profile of eprinomectin excretion in sheep faeces. AB - An analytical procedure has been introduced to enable study of the time profile of eprinomectin excretion in sheep faeces. Eprinomectin was extracted from sheep faeces with acetonitrile, the extract was cleaned by solid-phase extraction (SPE), and, after derivatization by reaction with N-methylimidazole, trifluoroacetic anhydride, and acetic acid, eprinomectin was analysed by high performance liquid chromatography (HPLC) with fluorescence detection. The method has a low detection limit (1.0 ng g(-1) of moist sheep faeces), a low quantification limit (2.5 ng g(-1) of moist sheep faeces), good recovery (in the range 78.8 to 87.1%), and good reproducibility (RSD<10%). The method was used to study the time-profile of excretion of eprinomectin in sheep faeces after a single topical administration of 0.5 mg kg(-1) b.w. of the drug. Because of its good recovery, precision, and sensitivity, the method has also proved applicable to further ecotoxicological studies of eprinomectin. PMID- 17109135 TI - Post streptococcal acute glomerulonephritis secondary to sporadic Streptococcus equi infection. AB - Streptococcus equi subspecies zooepidemicus infection is rare in humans, but a well-known cause of pyogenic disease in cows and horses. S. zooepidemicus uncommonly causes post-strep glomerulonephritis (PSGN) in humans via epidemic outbreaks. We present a sporadic case of post S. zooepidemicus glomerulonephritis in a child most probably contracted from a horse. The 14-year-old girl presented with the typical signs of PSGN, with S. equi zooepidemicus isolated from a blood culture, together with a low C3 and raised anti-DNAse B. This is the first known report of a sporadic case of PSGN in a child caused by this organism. PMID- 17109136 TI - Role of kidney chloride channels in health and disease. AB - Chloride channels are expressed along the entire mammalian nephron. They participate in transepithelial chloride transport, cell volume regulation and acidification of intracellular vesicles. Some chloride channels are constitutively active and others are regulated by either second messengers such as cAMP or Ca(++) or secondary to changes in membrane potential. The molecular identities of a number of chloride channels within the kidney are still unknown. Abnormalities in chloride channel expression and function in the kidney can cause a range of disorders such as autosomal recessive Dent's disease, Bartter's syndrome, renal tubular acidosis and diabetes insipidus. The purpose of this review is to give an overview of the chloride channels in the kidney and to focus on the function of renal chloride channels as revealed by diseases associated with channel dysfunction. PMID- 17109137 TI - Intraperitoneal treatment with darbepoetin for children on peritoneal dialysis. AB - To determine the efficacy and safety of intraperitoneal administration of darbepoetin in children with renal anemia on peritoneal dialysis, we conducted a single-arm, retrospective, two-centre study in which children were treated with intraperitoneal darbepoetin at the end of nightly intermittent peritoneal dialysis. Controls were those children treated with intraperitoneal erythropoietin six months before conversion to darbepoetin. Children were converted with the conversion factor 200 units erythropoietin=1 microg darbepoetin. Children who started with darbepoetin, started with 0.45 microg/kg/week. Nineteen children entered the study. The mean age was 6.8 years. Eight children were converted from 201 U/kg/week intraperitoneal erythropoietin to 1.0 microg/kg/week intraperitoneal darbepoetin. They were treated for a median period of 31.5 months. Median darbepoetin dose for an adequate erythropoesis over this period was 0.79 microg/kg/week. All 19 children were treated with darbepoetin for a median period of 13.4 months. The median dose for an adequate erythropoesis over this period was 0.63 microg/kg/week. The peritonitis incidence during this study was once every 25.1 months. Three children developed hypertension; one child developed headache. These complications developed after a rapid increase of hemoglobin concentration. Intraperitoneal administration of darbepoetin is effective and safe for children on peritoneal dialysis. PMID- 17109138 TI - Pattern of double glomerulopathy in children. AB - Occasional case reports have been issued on children with double glomerulopathy, involving either the coexistence of two different glomerulopathies or superimposition of a second glomerulopathy onto a first. A retrospective clinicopathological review of 294 children who had received renal biopsies resulted in 9 (3.1%) being confirmed to have double glomerulopathy. Superimposed glomerulopathy was diagnosed by a second renal biopsy in two cases, and coexistence of two glomerulopathies was confirmed by single biopsy in seven. Original glomerulopathies were those with a chronic course, such as Alport syndrome, IgA nephropathy, relapsing minimal-change nephrotic syndrome, Frasier syndrome, and thin basement membrane nephropathy. The superimposing glomerulopathies were common types in children, such as postinfectious glomerulonephritis, IgA nephropathy, and Henoch-Schonlein nephritis. Thus, the pattern of double glomerulopathy was considered to be due to the chance occurrence of two different glomerulopathies without a common pathogenesis. Acute nephritic symptoms of superimposed glomerulopathies resolved almost completely during follow-up in most cases. Double glomerulopathies are not rare in children and may occur by chance alone in most cases. The possibility of superimposed glomerulopathy should be suspected if the clinical course of a glomerulopathy changes atypically. However, the long-term influence of a superimposed glomerulopathy on renal functional deterioration remains unclear. PMID- 17109139 TI - Ca2+-Mg2+-dependent ATP-ase activity and calcium homeostasis in children with chronic kidney disease. AB - Extracellular calcium concentrations in humans are thousands times higher than within cells. Maintenance of such gradient requires specific regulation including intracellular stores, Ca binding proteins and transmembrane protein systems. The aim of the study was to estimate PMCA (plasma membrane Ca-transporting adenosine triphosphatase; ATPase 3.6.1.38) activity and calcium homeostasis in erythrocytes of children with chronic kidney disease (CKD). Twenty-one children wth CKD stages 1-3 (group I) and 18 healthy children (group II) were examined. Group I was divided into two subgroups: Ia (8 patients with normal intact parathyroid hormone, iPTH, serum levels) and Ib (13 patients with increased iPTH). iPTH, urea, creatinine, inorganic phosphorus, cytosolic Ca2+ in red blood cells (R-Ca), and PMCA were determined. Significantly elevated R-Ca levels were observed in children from subgroup Ib in comparison with group II and subgroup Ia. The lowest activity of PMCA was found in subgroup Ia and Ib in comparison with group II. There was a negative correlation between PMCA and R-Ca in group Ia and Ib (r= 0.8, r=-0.9, respectively). In children with CKD treated conservatively, activity of PMCA in erythrocytes is disturbed. An increase in R-Ca and decrease in PMCA activity are also observed. PMID- 17109140 TI - Therapeutic approach to FSGS in children. AB - Therapy of primary focal segmental glomerulosclerosis (FSGS) in children incorporates conservative management and immunosuppression regimens to control proteinuria and preserve kidney function. In long-term cohort studies in adults and children with primary FSGS, renal survival has been directly associated with degree of proteinuria control. This educational article reviews the current therapeutic approach toward children with primary FSGS. PMID- 17109141 TI - Effect of selenium-deficient diet on tubular epithelium in normal rats. AB - Selenium (Se) deficiency reduces glutathione peroxidase (GPx) activity, resulting in increased oxidative stress. We examined how Se deficiency induces renal injury via oxidative stress over time during the Se-deficient period. Seventy-two male Wistar rats were divided into two groups and fed either a control or Se-deficient diet. Rats were sacrificed on weeks 1, 2, 4, 6, 9, and 12. Blood and urine samples were collected, and the kidneys were removed. Urinalysis was performed, and creatinine clearance (Ccr) was calculated. Expressions of cellular GPx (cGPx) and phospholipid hydroperoxidase GPx (PHGPx) mRNA and GPx activity were measured. Histology was evaluated by light microscopy with immunohistochemistry for 4 hydroxy-2-nonenal (HNE) and vimentin. The Se-deficient diet caused significant decreases in GPx activity and cGPx mRNA expression but no change in PHGPx mRNA, together with significant proteinuria and glucosuria and slight decline in Ccr. The Se-deficient diet induced calcification in the kidney and increased the distribution of HNE and vimentin immunostaining in proximal tubuli, particularly around the outer medulla stripe. However, the histological damage did not progress after 6 weeks of deficiency. Se deficiency induces proteinuria and glucosuria with renal calcification, which may be primarily induced by injury of proximal tubuli via oxidative stress. PMID- 17109142 TI - Hypoxia/re-oxygenation injury induces apoptosis of LLC-PK1 cells by activation of caspase-2. AB - Hypoxia/re-oxygenation injury induces apoptosis in renal tubule cells, but its underlying molecular pathways are not fully elucidated. Activation of caspase-2 has recently been proposed as a novel mechanism of apoptosis in fibroblasts. In this study we examined whether hypoxia/re-oxygenation injury induces apoptosis in proximal tubule cells by activation of caspase-2. Porcine proximal tubule (LLC PK1) cells were subjected to hypoxia/re-oxygenation injury in the presence or absence of caspase inhibitors. Apoptosis was detected by DNA laddering, flow cytometry, and immunocytochemistry for Bax and cytochrome c. The activity of caspases-2, 8 and 9 was measured. Apoptosis was evident after hypoxia/re oxygenation and was best prevented by pretreatment with caspase-2 inhibitor. Hypoxia/re-oxygenation resulted in a dramatic increase in caspase-2 activity (32 fold, in comparison with a 16-fold increase in caspase-8 activity and a tenfold increase in caspase-9 activity). Immunocytochemistry revealed Bax activation and translocation to mitochondria and cytochrome c release into the cytosol following hypoxia/re-oxygenation, both of which were significantly suppressed by pretreatment with caspase-2 inhibitor. These results indicate that hypoxia/re oxygenation injury in cultured proximal tubule cells induced apoptosis by activation of caspase-2, which is required for the mitochondrial translocation of Bax. PMID- 17109143 TI - Cadmium effects on populations of root nuclei in two pea genotypes inoculated or not with the arbuscular mycorrhizal fungus Glomus mosseae. AB - Plants possess a broad range of strategies to cope with cadmium (Cd) stress, including the arbuscular mycorrhizal (AM) symbiosis. In cell responses towards Cd, the contribution of changes in ploidy levels is still unclear. We used flow cytometry to investigate if nuclear ploidy changes are involved in response mechanisms toward Cd and to analyze the effect of the symbiotic status on populations of nuclei. The impact of Cd was investigated in roots of two pea (Pisum sativum L.) genotypes differing in their Cd-sensitivity (Cd-sensitive VIR4788 and Cd-tolerant VIR7128). In pea seedlings grown under hydropony, 25 and 250 microM Cd concentrations lead to an increase in 4 C together with a decrease in 2 C nuclei. The same genotypes, grown in soil/sand substrate, were inoculated or not with the AM fungus Glomus mosseae BEG12 and treated or not with Cd at transplanting (Cd1) or 2 weeks after (Cd2). The Cd2 increased the proportion of 6 and 8 C nuclei in the mycorrhizal VIR4788 and in the non-mycorrhizal VIR7128 genotypes. Thus, changes in ploidy levels reflect pea responses towards Cd, which are modulated by the symbiotic interaction. The Cd-induced increase in ploidy may account for changes in DNA transcription and/or translation. PMID- 17109144 TI - Mycorrhizal and dark septate endophytic fungi of Pedicularis species from northwest of Yunnan Province, China. AB - Colonization of mycorrhizal and root endophytic fungi in 14 Pedicularis species from northwest of Yunnan Province, southwest China, was examined. These species included: Pedicularis gracilis Wall., Pedicularis longipes Maxim., Pedicularis axillaris Franch., Pedicularis cephalantha Franch., Pedicularis tenuisecta Franch., Pedicularis tapaoensis Tsoong, Pedicularis likiangensis Franch., Pedicularis dichotoma Bonati, Pedicularis yui Li, Pedicularis rhinanthoides Schrenk, Pedicularis rex C.B. Clarke, Pedicularis longiflora Rudolph., Pedicularis siphonantha Don, and Pedicularis oxycarpa Franch., among which nine are endemic to China (one to Yunnan). Three types of potentially beneficial fungi associated with roots of these species were observed, namely, arbuscular mycorrhizal fungi, ectomycorrhizal fungi, and dark septate endophytic fungi (DSEF), with DSEF as the most common colonizers. An unexpected high colonization level was detected in this hemiparasitic genus. Of the 19 sampling sites examined, 10 gave colonization frequency of above 50% and 6 showed a colonization index of above 50. Heavy colonization suggested a significant ecological role of these fungi and their potential to be applied to successful cultivation of these intractable plants. PMID- 17109145 TI - Arbuscular mycorrhizal morphology and dark septate fungal associations in medicinal and aromatic plants of Western Ghats, Southern India. AB - We investigated roots of 107 medicinal and aromatic plants (MAPs) in the Western Ghats region of Southern India for arbuscular mycorrhizal (AM) and dark septate endophyte (DSE) associations. Of the 107 MAPs belonging to 98 genera in 52 families examined, 79 were AM and 38 harbored a DSE association. Typical Arum- and Paris-type mycorrhizas are first reported in the presumed nonmycorrhizal family Amaranthaceae. Similarly, DSE associations are recorded for the first time in nine plant families and 37 plant species. Thirty MAPs had both AM and DSE associations. The number of MAPs having Arum-type mycorrhiza was greater than those having Paris-type. This was more prominent among herbaceous plants than in trees where the Paris-type was predominant. Similarly, the Arum-type was more prevalent in annuals than in perennials. DSE associations were more frequent in herbs and perennials compared to other MAPs. PMID- 17109146 TI - Tests of stationarity and cointegration of health care expenditure and gross domestic product: an application to Turkey. AB - This study examines the long-run relationship among the per capita private, public, and total health care expenditure and per capita gross domestic product and population growth of Turkey. We find some evidence of multivariate cointegrating relationships among the health care expenditure and gross domestic product, and population growth. We further find a bivariate cointegrating relationship between private health care expenditure and per capita gross domestic product. Accordingly, a 10% increase in gross domestic product would translate into a 21.9% increase in total health care expenditure while controlling population growth. The income elasticity of health expenditure is found to be greater than 1, implying that health care is a luxury good in Turkey. Finally we note that there exists one-way causality running from per capita gross domestic product to various definitions of health care expenses. PMID- 17109147 TI - Changes in extracellular pH are neither required nor sufficient for activation of mitogen-activated protein kinases (MAPKs) in response to systemin and fusicoccin in tomato. AB - Leaf wounding and the wound signaling peptide systemin induce expression of wound response genes while the fungal toxin fusicoccin (FC) induces expression of pathogenesis-related genes. Consistent with their functional differences, FC and systemin regulate the extracellular pH in opposite ways, with systemin inducing an alkalinization and FC an acidification response. Here we show that systemin, wounding and FC activate the same mitogen-activated protein kinases (MAPKs; MPKs) MPK1 and 2 in tomato (Lycopersicon esculentum) leaves and L. peruvianum suspension-cultured cells. Wounding and FC activated an additional MAPK, MPK3. Pronounced differences were observed with regard to MAPK activation kinetics. FC induced prolonged, and systemin transient activity of the MAPKs. This shows that functionally different elicitors engage the same signaling components, yet induce signal-specific activation dynamics. A comparative analysis of pH effects and MAPK activity in response to specific treatments revealed that the kinetics of pH changes and MAPK activation did not correlate. Simultaneous application of FC and systemin did not lead to immediate pH changes but resulted in rapid increases in MAPK activity. Furthermore, changes in extracellular pH could be induced without concomitant MAPK activation by exchanging conditioned medium with fresh medium. This shows that changes in the extracellular pH are neither required nor sufficient for MAPK activation, suggesting that signaling pathways involving MAPKs and extracellular pH changes operate in parallel and are not part of the same linear pathway. PMID- 17109148 TI - MIR166/165 genes exhibit dynamic expression patterns in regulating shoot apical meristem and floral development in Arabidopsis. AB - The miR166/165 group and its target genes regulate diverse aspects of plant development, including apical and lateral meristem formation, leaf polarity, and vascular development. We demonstrate here that MIR166/165 genes are dynamically controlled in regulating shoot apical meristem (SAM) and floral development in parallel to the WUSCHEL (WUS)-CLAVATA (CLV) pathway. Although miR166 and miR165 cleave same target mRNAs, individual MIR166/165 genes exhibit distinct expression domains in different plant tissues. The MIR166/165 expression is also temporarily regulated. Consistent with the dynamic expression patterns, an array of alterations in SAM activities and floral architectures was observed in the miR166/165-overproducing plants. In addition, when a MIR166a-overexpressing mutant was genetically crossed with mutants defective in the WUS-CLV pathway, the resultant crosses exhibited additive phenotypic effects, suggesting that the miR166/165-mediated signal exerts its role via a distinct signaling pathway. PMID- 17109149 TI - Chemical composition of epicuticular wax crystals on the slippery zone in pitchers of five Nepenthes species and hybrids. AB - Plants of the carnivorous genus Nepenthes efficiently trap insects in leaf pitchers, mostly employing epicuticular wax crystals on the pitcher walls to make them slippery for the prey. In the present study, the compositions and micromorphologies of the wax crystals of five Nepenthes species and hybrids were analysed in order to test whether the chemical principles underlying this ecological function are widespread within the genus. Three wax layers could be distinguished within the Nepenthes pitcher cuticles: (1) the outermost part of the crystals forming the platelets visible in standard scanning electron microscopy, (2) the bottom portion of the epicuticular wax crystals, and (3) an intracuticular wax layer. The composition of the intracuticular wax differed significantly from that of the neighbouring epicuticular layer. The compositions of corresponding wax mixtures from all five Nepenthes species and hybrids were very similar, with almost equal amounts of very long chain aldehydes and primary alcohols. While triacontanal (C(30) aldehyde) was prevailing in the epicuticular crystals of Nepenthes albomarginata and Nepenthes x intermedia, Nepenthes x superba and Nepenthes x henriana were found to have especially high percentages of dotriacontanal (C(32) aldehyde). Nepenthes "khasiana" had an intermediate aldehyde composition with almost equal amounts of both chain lengths. PMID- 17109150 TI - Biphenyl synthase, a novel type III polyketide synthase. AB - Biphenyls and dibenzofurans are the phytoalexins of the Maloideae, a subfamily of the economically important Rosaceae. The carbon skeleton of the two classes of antimicrobial secondary metabolites is formed by biphenyl synthase (BIS). A cDNA encoding this key enzyme was cloned from yeast-extract-treated cell cultures of Sorbus aucuparia. BIS is a novel type III polyketide synthase (PKS) that shares about 60% amino acid sequence identity with other members of the enzyme superfamily. Its preferred starter substrate is benzoyl-CoA that undergoes iterative condensation with three molecules of malonyl-CoA to give 3,5 dihydroxybiphenyl via intramolecular aldol condensation. BIS did not accept CoA linked cinnamic acids such as 4-coumaroyl-CoA. This substrate, however, was the preferential starter molecule for chalcone synthase (CHS) that was also cloned from S. aucuparia cell cultures. While BIS expression was rapidly, strongly and transiently induced by yeast extract treatment, CHS expression was not. In a phylogenetic tree, BIS grouped together closely with benzophenone synthase (BPS) that also uses benzoyl-CoA as starter molecule but cyclizes the common intermediate via intramolecular Claisen condensation. The molecular characterization of BIS thus contributes to the understanding of the functional diversity and evolution of type III PKSs. PMID- 17109151 TI - Proline-rich cell wall proteins accumulate in growing regions and phloem tissue in response to water deficit in common bean seedlings. AB - Plant cell walls undergo dynamic changes in response to different environmental stress conditions. In response to water deficit, two related proline-rich glycoproteins, called p33 and p36, accumulate in the soluble fraction of the cell walls in Phaseolus vulgaris (Covarrubias et al. in Plant Physiol 107:1119-1128, 1995). In this work, we show that p33 and p36 are able to form a 240 kDa oligomer, which is found in the cell wall soluble fraction. We present evidence indicating that the highest accumulation of these proteins in response to water deficit occurs in the growing regions of common bean seedlings, particularly in the phloem tissues. These proteins were detected in P. vulgaris cell suspension cultures, where the p33/p36 ratio was higher under hyperosmotic conditions than in bean seedlings subjected to the same treatment. The results support a role for these proteins during the plant cell response to changes in its water status, and suggest that cell wall modifications are induced in active growing cells of common bean in response to water limitation. PMID- 17109152 TI - Synovial tumefactive extramedullary hematopoiesis associated to polycythemia vera. AB - The case of a 66-year-old male patient with a chronic myeloproliferative type polycythemia vera disorder, who after 2 years of evolution is developing a tumefactive extramedullary hematopoiesis (TEH) located in the synovial of the articulation in the right knee, is described. The tumor histologically consists of a relatively lax and edematous synovial structure diffusely infiltrated by mature and semimature hematopoietic cellular population. The simultaneous study of the bone marrow reveals medullar spaces full of hematopoietic cellularity, with a predominance of megakaryocytic and red series, and with the addition of severe reticulin fibrosis, facts that suggest a progression toward myelofibrosis. The TEH developed in tissues without a reticulum endothelial system is very uncommon. We provide data about the first case located in the synovial membrane and we review the literature regarding this pathologic entity. PMID- 17109153 TI - Primary intravascular myxoid leiomyosarcoma of the femoral vein presenting clinically as deep vein thrombosis: a case report. PMID- 17109154 TI - Reply: mixed germ cell sex cord-stromal tumors of the testis and ovary. PMID- 17109155 TI - Intraocular triamcinolone acetonide for macular edema due to CRVO. A multifocal ERG and OCT study. AB - PURPOSE: To investigate the effectiveness of intravitreal triamcinolone acetonide in the treatment of cystoid macular edema due to central retinal vein occlusion (CRVO). A noncomparative, prospective, interventional case series. METHODS: In this study 15 eyes of 15 patients (9 males and 6 females) with macular edema due to non-ischemic CRVO were treated with intravitreal injection of 4 mg triamcinolone acetonide and followed-up for 1 year. Examination included measurement of best-corrected visual acuity (BCVA) for distance, measurement of intraocular pressure (IOP), fluorescein angiography, foveal retinal thickness measurement by optical coherence tomography (OCT), and multifocal electroretinography recordings (MFERG) preoperatively 3, 6 and 12 months postoperatively. RESULTS: The visual acuity increased to a significant degree at 3 months, to a smaller degree at 6 months but at 12 months there was no significant improvement. The OCT macula thickness improved to a significant level all the follow-up period but with less significance at 12 months. The MFERG recordings from the fovea showed significant improvement at 3 and 6 months. The MFERG from the parafovea area showed significant improvement at 3 and 6 and to a smaller degree at 12 months. The intraocular pressure increased at 3 and 6 months but returned to pretreatment level at 12 months. CONCLUSION: Intravitreal injection of triamcinolone acetonide leads to a significant improvement of mean VA in patients with macular edema due to CRVO. However, this significant effect is not permanent and persists for a maximum of 3-6 months. Thereafter all the indexes tend to deteriorate. PMID- 17109156 TI - A teenager with nightblindness and cystic maculopathy: enhanced S cone syndrome (Goldmann-Favre syndrome). AB - Poor night vision and pigmentary retinopathy are hallmarks of retinitis pigmentosa, but variations in the type of pigmentation and in the characteristics of the electroretinogram (ERG) can point to other diagnoses. The case below (enhanced S cone syndrome) is a good example of this, and demonstrates the diagnostic importance of clinical electrophysiology. PMID- 17109157 TI - ISCEV Standard for Clinical Electro-oculography (EOG) 2006. AB - The Clinical Electro-oculogram (EOG) is an electrophysiological test of function of the outer retina and retinal pigment epithelium (RPE) in which the change in the electrical potential between the cornea and the ocular fundus is recorded during successive periods of dark and light adaptation. This document sets out a Standard Method for performance of the test, and also gives detailed guidance on technical and practical issues, and on reporting test results. The main object of the Standard is to promote consistent quality of testing and reporting within and between centres. This 2006 Standard, from the International Society for Clinical Electrophysiology of Vision (ISCEV: www.iscev.org ), is a revision of the previous Standard published in 1993, and reviewed and re-issued in 1998. PMID- 17109158 TI - Pediatric clinical visual electrophysiology: a survey of actual practice. AB - PURPOSE: Survey the actual clinical practice of pediatric visual electrophysiology. The electrophysiologists surveyed were members of the International Society for Clinical Electrophysiology of Vision (ISCEV). METHODS: A self-administered questionnaire with 55 items about visual evoked potential (VEP) and electroretinogram (ERG) testing of pediatric patients was sent to ISCEV members. The survey queried personnel, facilities, referral patterns and conduct of tests. RESULTS: Nearly all respondents (94%) had advanced scientific or clinical degrees or both, and most (96%) worked in academic or medical facilities. Of the 71 respondents, 68 tested patients 12 years or younger, and nearly all of those performed both VEPs and ERGs. However, fewer than a third did high volume (>10/month) testing of infants and young children (< or =6 years). Eye care professionals and neurologists made the majority (57%) of the referrals, with the most common reason for referral being suspected visual impairment. Conduct of a pediatric test session often required more than one practitioner. For both VEP and ERG, more than 70% of respondents required at least 30 min for each test. The majority indicated that they followed the ISCEV standards for stimuli and data acquisition. Almost all (94%) reported using the ISCEV recommended VEP electrode configuration. For ERG, most (88%) used ocular contact electrodes (including contact lens, thread, foil and HK loop), but 12% used skin electrodes exclusively and some (17%) used skin electrodes at times. CONCLUSIONS: Pediatric ERG and VEP testing is a labor intensive endeavor of highly trained professionals. ISCEV technical standards are typically met or exceeded, indicating that high quality testing of infants and children is feasible. Revision of the ISCEV ERG standard is necessary to bring actual practice into accord with evidence-based recommendations for infant testing. PMID- 17109159 TI - The influence of luminance on the multifocal ERG. AB - PURPOSE: To assess the efficacy of high luminance in increasing the amplitude of the multifocal electroretinogram (mfERG). We examined 5 male and 5 female volunteers in the age of 22-52 years (median 28 years). Three different stimulus luminance levels were applied: the bright areas of the stimulus pattern were set to 150, 300 and 500 cd/m(2). We recorded the potentials via DTL electrodes using the VERIS Science 4.4 system with 61 hexagons, pupils were dilated. Analysis was based on the 5 ring averages. RESULTS: Across all hexagons and subjects, the response density (approximately amplitude) rose by 20% when increasing the luminance by a factor of 3.3. The peak times decreased slightly with higher luminance, by less then 1.5 ms. CONCLUSIONS: Combining the present results with those from two previous studies, the gain (= relative amplitude increase for relative luminance increase) is close to 0.4 over a range of 56-700 cd/m(2). The stimulus luminance range suggested in the mfERG guidelines seems well chosen. PMID- 17109160 TI - Development of species identification tests targeting the 16S ribosomal RNA coding region in mitochondrial DNA. AB - This report describes the development of a species testing system based on the diversity of nucleotide sequences in mitochondrial DNA (mtDNA) among species. Five species, human, cow, pig, dog, and cat, were considered. The partial nucleotide sequences in 16S ribosomal RNA coding region were chosen as the target for discriminating the species. The sequence diversities of this approximately 400 bp long region ranged from 15.7 to 24.1% among the five species. Sequencing analysis of this target on 50 individuals of each species (53 for dogs) revealed that the nucleotide sequences were well preserved within species. Species specific PCR for each species was also designed, and satisfactory results with regard to both sensitivity and specificity were obtained. A validation study with DNA extracted from bovine bone exposed to the environment revealed that the PCRs designed in this study worked correctly. From the results obtained, it was shown that this testing system could be a good tool for species identification. One successful case report is also demonstrated. PMID- 17109161 TI - Growth kinetics and Pho84 phosphate transporter activity of Saccharomyces cerevisiae under phosphate-limited conditions. AB - The effect of phosphate (P ( i )) concentration on the growth behavior of Saccharomyces cerevisiae strain CEN.PK113-5D in phosphate-limited batch and chemostat cultures was studied. The range of dilution rates used in the present study was 0.08-0.45 h(-1). The batch growth of yeast cells followed Monod relationship, but growth of the cells in phosphate-limited chemostat showed change in growth kinetics with increasing dilution rates. The difference in growth kinetics of the yeast cells in phosphate-limited chemostat for dilution rates below and above approximately 0.2 h(-1) has been discussed in terms of the batch growth kinetic data and the change in the metabolic activity of the yeast cells. Immunological detection of a C-terminally myc epitope-tagged Pho84 fusion protein indicated derepressive expression of the Pho84 high-affinity P ( i ) transporter in the entire range of dilution rates employed in this study. Phosphate transport activity mediated by Pho84 transporter was highest at very low dilution rates, i.e. 0.08-0.1 h(-1), corresponding to conditions in which the amount of synthesized Pho84 was at its maximum. PMID- 17109162 TI - Erythroderma in a 1-month-old boy. PMID- 17109163 TI - Characteristics of tinnitus in childhood. AB - Despite its incidence, there are still few reports in literature relating to tinnitus in children. Almost all data were collected by means of questionnaires or in a limited population of children. In order to collect data in a homogeneous way and directly from the patients, the protocol of study proposed by Savastano has been applied to 1,100 children. The results showed tinnitus as present in 374 children but only 6.5% of the cases complained spontaneously about it. In all, 76.4% of the children demonstrated normal hearing, whereas 64.5% reported being bothered by their tinnitus. Tinnitus measurements were obtained and are reproducible in all patients older than 8 years of age. The loudness level was <10 dB in 48.6% of cases, which was higher than 10 dB in 51.4%. As for the frequency distribution, in most cases it appears to be between 0 and 1,000 Hz. There is a correspondence between the loudness level and masking level. A total inhibition of <60 s for most children with lower loudness was obtained. The present study demonstrates that the application in the infancy of a specific protocol of study allows the presence of tinnitus to be discovered, giving specific and detailed information about it so as to minimize its damage to be obtained. Moreover, for the first time, data regarding the measurement of tinnitus in childhood has been gathered. PMID- 17109164 TI - Assessment of muscle shortening and static posture in children with persistent asthma. AB - Asthmatic patients experience an increase in airway resistance that overburdens both respiratory and non-respiratory muscles. The objective of the present study was to determine whether children with persistent asthma present muscle shortening and postural changes. The 60 boys evaluated, aged 7-12 (pubertal ages up to Tanner stage G2) were divided into three age- and BMI-matched groups of equal number: CON (no history of asthma or allergy); MPA (mild persistent asthma); SPA (severe persistent asthma). Pulmonary function, muscle shortening and static posture were evaluated. The SPA group presented higher protraction of the head and shoulder compared with the CON group [9.5 (6.0-12.0) degrees vs 5.5 (0.0-12.0) degrees, P < 0.05; 0.89 (0.80-0.94) anterior/posterior ratio vs 0.94 (0.87-1.1) anterior/posterior ratio, P < 0.01)]. Severe asthmatic patients also presented shortening of arm flexor and posterior muscle of the thigh compared with the CON group [18.0 (10.0-24.0) degrees vs 12.0 (6.0-16.0) degrees, P < 0.05; and 16.5 (10.0-38.5) cm vs 8.0 (0.0-21.0) cm, respectively, P < 0.05]. Chest expansion at axillar and xiphoid levels were limited in SPA subjects compared with CON subjects [3.7 (1.5-6.5) cm vs 5.5 (2.0-8.0) cm and 4.7 (1.5 6.5) vs 6.0 (3.5-8.0) cm, respectively, P < 0.01]. SPA subjects also presented a higher incidence of lumbar spine straightening compared with CON and MPA subjects. Moderate asthmatic subjects presented intermediate values compared with severe and control subjects in five out nine evaluated outcomes. Our data suggest that severe asthmatic children present postural adaptations and muscle shortening that seem to be related to disease severity. PMID- 17109165 TI - Concordance between school outcomes and developmental follow-up results of very preterm and/or low birth weight children at the age of 5 years. AB - INTRODUCTION: Long-term follow-up studies have revealed a high frequency of developmental disturbances in preterm survivors of neonatal intensive care who were formerly considered to be non-disabled. These developmental disturbances interfere with the acquisition of everyday skills and, in particular, with normal school functioning. METHODS: Developmental and school outcomes of 355 children, age 5 years at the time of the study, who had a mean gestational age of 30.2 weeks (SD: 1.95) and a mean birth weight of 1272 g (SD: 326) were investigated. Children with severe handicaps were excluded from the study. Perinatal data, information from a parental and school questionnaire and data from standardized developmental tests were used to explain the differences. RESULTS: An agreement of 72% was found between developmental follow-up and school outcomes. Normal developmental results but problematic school outcomes were found for 15% of the children tested. There were more boys than girls in this latter group as well as small-for-gestational-age children with relatively poor motor or language development. The schools had not identified problems in 13% of the children, whereas their developmental outcomes were problematic. These children had less neonatal morbidity and relatively higher IQ's than children who also had problematic developmental outcomes but who had been signalled as problematic by their schools. CONCLUSIONS: Schools have a good insight in the school functioning of children who are developing well and of children with the lowest developmental scores and the most complicated neonatal histories. How school and developmental outcomes interrelate in the in-between groups remains a challenging question that could be answered by following these children throughout their school career. PMID- 17109166 TI - Lower abdominal inflammatory myofibroblastic tumor -an unusual presentation- a case report and brief literature review. AB - A 9-year-old girl presented with lethargy, malaise & chest pain. Her blood counts confirmed hypochromic microcytic anemia. She was prescribed iron supplements. Subsequently she was admitted to our hospital with fever and increasing chest and abdominal pain. She was treated with antibiotics, and a diagnosis of "early chest infection" was made. Over the following 2 weeks she failed to improve, and her anemia worsened. She was readmitted, and found to have a mass in her lower abdomen with pressure symptoms on her bowel and bladder. A white-cell scan showed increased uptake in right lower quadrant. An ultrasound and a CT scan confirmed a mass adjacent to her bladder. Needle biopsy showed it to be an unusual localization of an inflammatory myofibroblastic tumor (IMT) of cecum. A presentation with chest pain, fever, anemia and pressure symptoms was highly unusual of a lower abdominal IMT mass. She had a successful excision of the tumor, with resolution of her symptoms. PMID- 17109167 TI - Exchange transfusion in a preterm infant with hyperbilirubinemia, staphylococcal scalded skin syndrome (SSSS) and sepsis. PMID- 17109168 TI - Administering radioiodine treatment; alternatives to nuclear medicine prescribing. PMID- 17109169 TI - Does (124)Iodo-deoxyuridine measure cell proliferation in NSCLC? Initial investigations with PET imaging and radio-metabolite analysis. PMID- 17109170 TI - Specific single-cell isolation and genomic amplification of uncultured microorganisms. AB - We in this study describe a new method for genomic studies of individual uncultured prokaryotic organisms, which was used for the isolation and partial genome sequencing of a soil archaeon. The diversity of Archaea in a soil sample was mapped by generating a clone library using group-specific primers in combination with a terminal restriction fragment length polymorphism profile. Intact cells were extracted from the environmental sample, and fluorescent in situ hybridization probing with Cy3-labeled probes designed from the clone library was subsequently used to detect the organisms of interest. Single cells with a bright fluorescent signal were isolated using a micromanipulator and the genome of the single isolated cells served as a template for multiple displacement amplification (MDA) using the Phi29 DNA polymerase. The generated MDA product was afterwards used for 16S rRNA gene sequence analysis and shotgun cloned for additional genomic analysis. Sequence analysis showed >99% 16S rRNA gene homology to soil crenarchaeotal clone SCA1170 and shotgun fragments had the closest match to a crenarchaeotal BAC clone previously retrieved from a soil sample. The system was validated using Methanothermobacter thermoautotrophicus as single-cell test organism, and the validation setup produced 100% sequence homology to the ten tested regions of the genome of this organism. PMID- 17109171 TI - Batch fermentations on synthetic mixed sugar and starch medium with amylolytic lactic acid bacteria. AB - The green crop drying industry in Denmark uses Italian rye grass, clover, and alfalfa as raw materials for the production of green pellets. The green crop drying industry solves its energy economical problems by heating and pressing of the green crop before drying. The produced sidestream is called brown juice. Brown juice was shown to be an excellent medium for lactic acid fermentation. The aim of this study was to investigate the utilisation of brown juice in the production of polylactic acid, where wheat starch would be added to increase the lactic acid yield and, thus, the feasibility of the process. A number of amylolytic lactic acid bacteria have been identified, and in this work, six different strains were tested for their ability to produce alpha-amylase and to utilise all sugars with high lactic acid yield in a medium with a complex composition of free sugars (brown juice) and starch. Lactobacillus plantarum A6 was the only strain that showed both a good lactic acid production and utilisation of starch in this medium. The growth rate of this strain was approximately 0.4 h(-1) and the lactic acid yield was 0.7. PMID- 17109172 TI - Role of an extracellular neutral protease in infection against nematodes by Brevibacillus laterosporus strain G4. AB - Proteases have been proposed as virulence factors in microbial pathogenicity against nematodes. However, what kinds of extracellular proteases from these pathogens and how they contribute to the pathogenesis of infections against nematode in vivo remain largely unknown. A previous analysis using a strain with a deletion in an extracellular alkaline protease BLG4 gene from Brevibacillus laterosporus demonstrated that BLG4 was responsible for the majority of nematicidal activity by destroying host's cuticle. In recent studies, a neutral protease NPE-4, purified from the mutant BLG4-6, was found to be responsible for the majority of the remaining EDTA-inhibited protease activity. However, the purified NPE-4 and recombinant NPE-4 in a related species Bacillus subtilis showed little nematicidal activity in vitro and were unable to degrade the intact cuticle of the host. It is interesting to note that the addition of NPE-4 improved the pathogenicity of crude enzyme extract from wild-type B. laterosporus but had no effect on the BLG4-deficient mutant. This result suggests that NPE-4 functions in the presence of protease BLG4. Moreover, NPE-4 could degrade proteins from the inner layer of purified cuticles from nematode Panagrellus redivivus in vitro. These results indicated that the two different bacterial extracellular proteases might play differential roles at different stages of infection or a synthetic role in penetration of nematode cuticle in B. laterosporus. This is among the first reports to systematically evaluate and define the roles of different bacterial extracellular proteases in infection against nematodes. PMID- 17109173 TI - A mannose-specific tetrameric lectin with mitogenic and antibacterial activities from the ovary of a teleost, the cobia (Rachycentron canadum). AB - A tetrameric lectin, with hemagglutinating activity toward rabbit erythrocytes and with specificity toward D-mannosamine and D(+)-mannose, was isolated from the ovaries of a teleost, the cobia Rachycentron canadum. The isolation protocol comprised ion exchange chromatography on CM-cellulose and Q-Sepharose, ion exchange chromatography by fast protein liquid chromatography (FPLC) on Mono Q, and finally gel filtration by FPLC on Superose 12. The lectin was adsorbed on all ion exchangers used. It exhibited a molecular mass of 180 kDa in gel filtration on Superose 12 and a single 45-kDa band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, indicating that it is a tetrameric protein. The hemagglutinating activity of the lectin was stable up to 40 degrees C and between pH 4 and pH 10. All hemagglutinating activity disappeared at 60 degrees C and at pH 1 and pH 13. The hemagglutinating activity was doubled in the presence of 0.1 microM FeCl3. The lectin exerted antibacterial activity against Escherichia coli with 50% inhibition at 250 microg. There was no antifungal activity toward Coprinus comatus, Fusarium oxysporum, Mycosphaerella arachidicola, and Rhizoctonia solani at a dose of 300 microg. The lectin exhibited maximal mitogenic response from mouse splenocytes at a concentration of 14 microM. PMID- 17109174 TI - [Structural femoral head allografts in revision surgery of loosened acetabular cups]. AB - BACKGROUND: Treatment of acetabular bone defects presents a great challenge in revision total hip arthroplasty (THA). Many methods of acetabular reconstruction have been described. The purpose of this study was to evaluate the midterm results of structural femoral head allografts for acetabular reconstruction. METHODS: Thirty-six patients (33 females and 3 males) with acetabular defects ranging from type 2C to type 3B according to Paprosky's classification were included in the study. In all cases acetabular defects were closed using allografts from femoral heads. In 13 cases an uncemented press-fit cup, in 17 cases a cemented polyethylene socket, and in 6 cases a Burch-Schneider antiprotrusion cage was implanted. The mean follow-up period was 84.2 months (range: 5-147). RESULTS: Four acetabular components failed. All 36 grafts were osseointegrated radiographically and formed a mechanically stable construction. The mean Harris Hip Score at the most recent follow-up was 79.8 points. The distance from the obturator line to the prosthesis head center was 3.73 cm (1.17 5.80 cm) preoperatively and 2.79 cm (0.85-4.8 cm) postoperatively (p<0.05). The distance from the teardrop figure to the prosthesis head center was 3.02 cm (1.0 5.8 cm) preoperatively and 3.25 cm (1.6-4.8 cm) postoperatively (p<0.001). CONCLUSIONS: Closure of acetabular defects of types 2C to 3B according to Paprosky's classification can be satisfactorily accomplished using femoral head allografts. These allografts may facilitate future revision surgery. Femoral heads are readily available due to widespread primary total hip replacement surgery. However, the use of structural femoral head allografts for acetabular reconstruction is cost intensive. Individual patient-related aspects, such as the function of revision arthroplasty, have to be considered when planning revision arthroplasty using femoral head allografts. PMID- 17109175 TI - [Reduction of traumatic dislocations and facet fracture-dislocations in the lower cervical spine]. AB - BACKGROUND: Traumatic facet dislocations and facet-fracture dislocations in the lower cervical spine (C2/C3 to C7/T1) are frequently associated with devastating neurological symptoms. A good outcome can only be achieved if the operator has wide and sound knowledge of reduction techniques and the best possible strategy is devised for the subsequent treatment of these severe lesions. PATIENTS AND METHODS: Between 1973 and 1997 a total of 117 of our patients met at least one of the following inclusion criteria: unilateral locked facet dislocation (48%), bilateral locked facet dislocations (23%), unilateral "perched" facet subluxation (14%), bilateral perched facet subluxation (12%), uni- or bilateral dislocation/perched subluxation with facet fractures (3%). RESULTS: Most of the lesions were located at the levels of C5/C6 and C6/7 (n=46 for each). Associated neurological deficits were present initially in 65% of patients: 35% had complete or incomplete spinal cord injuries (tetraplegia), 2% were paraplegic, and 28% had cervical radiculopathies. CONCLUSIONS: Closed reduction (e.g. with the aid of a halo ring) should be carried out as soon as possible after lower cervical spine dislocation or facet-fracture dislocation, as both the success rate of reduction and the potential for recovery from neurological deficits are clearly higher when reduction is achieved within the first 4 h after the initial injury. PMID- 17109176 TI - Influence of climate variability on whitefish (Coregonus lavaretus) year-class strength in a deep, warm monomictic lake. AB - The year-class strength (YCS) of Blaufelchen (Coregonus lavaretus) in deep Upper Lake Constance was analysed for a 52-year period, from 1947 to 1998. Despite strong anthropogenic influences on the species' population dynamics due to cultural eutrophication and oligotrophication, intense fishing, and large-scale stocking, the influence of climate variability associated with the North Atlantic Oscillation (NAO) is apparent in the data set. This influence is significant although large-scale stocking of cold-bred larvae was performed to avoid a mismatch of larvae with their food. The importance of stocking on YCS, however, is unclear and was only detectable when analysing a subset of the data. In addition to climate variability a yet unidentified factor related to zooplankton suitability as food for fish larvae, and density-dependent mortality probably related to cannibalism do significantly influence YCS. The NAO seemed to influence YCS twofold, through temperature effects on egg development time and on larval growth rate. The first of these two mechanisms is related to the NAO via a time lag of 1 year due to the specific mixing dynamics of warm monomictic Lake Constance. Hence, a warm winter in the year before spawning results in earlier hatching of larvae, that is, hatching is decoupled from the actual meteorological conditions. This should make the larvae very prone to mismatch the dynamics of their food. However, we found no evidence for such a mismatch in this 52-year data set. PMID- 17109177 TI - Production of phytoestrogen S-equol from daidzein in mixed culture of two anaerobic bacteria. AB - An anaerobic incubation mixture of two bacterial strains Eggerthella sp. Julong 732 and Lactobacillus sp. Niu-O16, which have been known to transform dihydrodaidzein to S-equol and daidzein to dihydrodaidzein respectively, produced S-equol from daidzein through dihydrodaidzein. The biotransformation kinetics of daidzein by the mixed cultures showed that the production of S-equol from daidzein was significantly enhanced, as compared to the production of S-equol from dihydrodaidzein by Eggerthella sp. Julong 732 alone. The substrate daidzein in the mixed culture was almost completely converted to S-equol in 24 h of anaerobic incubation. The increased production of S-equol from daidzein by the mixed culture is likely related to the increased bacterial numbers of Eggerthella sp. Julong 732. In the mixture cultures, the growth of Eggerthella sp. Julong 732 was significantly increased while the growth of Lactobacillus sp. Niu-O16 was suppressed as compared to either the single culture of Eggerthella sp. Julong 732 or Lactobacillus sp. Niu-O16. This is the first report in which two metabolic pathways to produce S-equol from daidzein by a mixed culture of bacteria isolated from human and bovine intestinal environments were successfully linked under anaerobic conditions. PMID- 17109178 TI - Multiple forearm diaphyseal fracture: reduction and plaster cast control at the end of growth. AB - The authors followed up 20 patients with multiple diaphyseal fractures of the radius and ulna who were treated nonoperatively and who healed with axial deviation >5 degrees in at least one plane 20.4+/-6.7 years after radiographic evidence of fracture union. Mean age at follow-up was 28.6+/-6.4 years. Radiographs were measured soon after reduction, at 10 days from reduction, at the end of treatment, and at follow-up (17/20). Both elbow and forearm range of motion (ROM) were compared with those of the contralateral side. At follow-up, ROM was normal and radiographs showed angular deviations <5 degrees . PMID- 17109179 TI - Excellent survival of pediatric dermatofibrosarcoma protuberans in Taiwanese. AB - The purpose of this study was to explore the clinical outcome of pediatric dermatofibrosarcoma protuberans (DFSP) in Taiwan. Patients with DFSP diagnosed before 18 years and treated in our institute from 1977 to 2002 were identified through institutional cancer registry. After approved by institutional research ethic committee, active patient contact and linkage with National Residency Registry were done to explore their clinical outcome (Clinical Trials gov Identifier: NCT00173355). Twelve patients were identified. Pathological review confirmed DFSP for all of them. All of them underwent wide excision and none of them underwent neoadjuvant or adjuvant therapy. Nine patients underwent operation as initial therapy and three of them underwent salvage therapy. We recognized only one uncontrollable local recurrence and subsequent death. The 10-year and 15 year overall survival rates were 100 and 83%, respectively, without obvious complications. The prognosis of pediatric DFSP in Taiwanese was excellent. PMID- 17109180 TI - [Unilateral segmental keratotic papules]. PMID- 17109181 TI - Surgical management of dural arteriovenous fistulas with transosseous arterial feeders involving the jugular bulb. AB - Dural arteriovenous fistulas located in the vicinity of the jugular foramen are complex vascular malformations and belong to the most challenging skull base lesions to treat. The authors comprehensively analyze multiple features in a series of dural arteriovenous fistulas with transosseous arterial feeders involving the jugular bulb. Four patients who underwent surgery via the transcondylar approach to treat dural arteriovenous fistulas around the jugular foramen were retrospectively reviewed. Previously, endovascular treatment was attempted in all patients. The success of the surgical treatment was examined with postoperative angiography. Complete obliteration of the dural arteriovenous fistulas (DAVFs) was achieved in three patients, and significant flow reduction in one individual. All patients had a good postoperative outcome, and only one experienced mild hypoglossal nerve palsy. Despite extensive bone drilling, an occipitocervical fusion was necessary in only one patient with bilateral lesions. The use of an individually tailored transcondylar approach to treat dural arteriovenous fistulas at the region of the jugular foramen is most effective. This approach allows for complete obliteration of the connecting arterial feeders, and removal of bony structures containing pathological vessels. PMID- 17109182 TI - Injecting drugs of abuse and immunity: implications for HIV vaccine testing and efficacy. AB - The recreational use of legal and illegal drugs has significant effects on immune responses and can potentially modulate susceptibility to infection by a number of pathogens. A number of agents including cannabinoids (marijuana), cocaine opiates, amphetamines, nicotine and alcohol were demonstrated to have potentially adverse effects on the susceptibility to infections, mediated most likely, by adverse effects on immunity. As such, these drugs of abuse could have significant and potentially adverse effects on the vaccination efficacy of a number of vaccines currently on the market and on potential experimental vaccines currently in the pipeline. This review will present an overview on how drugs of abuse potentially impacts immune responses and vaccination efficacy. The emphasis of this review will be the effects of opiate abuse, as exemplified by injecting/intravenous drug users (IDU), on HIV/AIDS and its potential impact on vaccine efficacy trials against this devastating infection/syndrome. PMID- 17109183 TI - Point sampling digital imagery with 'SamplePoint'. AB - Measuring percent occurrence of objects from digital images can save time and expense relative to conventional field measurements. However, the accuracy of image analysis had, until now, not reached the level of the best conventional field measurements. Additionally, most image-analysis software programs require advanced user training to successfully analyze images. Here we present a new software program, 'SamplePoint,' that provides the user a single-pixel sample point and the ability to view and identify the pixel context. We found SamplePoint to allow accuracy comparable with the most accurate field-methods for ground-cover measurements. Expert use of the program requires minimal training and its ease of use allows rapid measurements from image data. We recommend SamplePoint for calibrating the threshold-detection level of image-analysis software or for making direct measurements of percent occurrence from digital images. PMID- 17109184 TI - Compensatory vs. pseudocompensatory evolution in molecular and developmental interactions. AB - The evolution of molecules, developmental circuits, and new species are all characterized by the accumulation of incompatibilities between ancestors and descendants. When specific interactions between components are necessary at any of these levels, this requires compensatory coevolution. Theoretical treatments of compensatory evolution that only consider the endpoints predict that it should be rare because intermediate states are deleterious. However, empirical data suggest that compensatory evolution is common at all levels of molecular interaction. A general solution to this paradox is provided by plausible neutral or nearly neutral intermediates that possess informational redundancy. These intermediates provide an evolutionary path between coadapted allelic combinations. Although they allow incompatible end points to evolve, at no point was a deleterious mutation ever in need of compensation. As a result, what appears to be compensatory evolution may often actually be "pseudocompensatory." Both theoretical and empirical studies indicate that pseudocompensation can speed the evolution of intergenic incompatibility, especially when driven by adaptation. However, under strong stabilizing selection the rate of pseudocompensatory evolution is still significant. Important examples of this process at work discussed here include the evolution of rRNA secondary structures, intra- and inter-protein interactions, and developmental genetic pathways. Future empirical work in this area should focus on comparing the details of intra- and intergenic interactions in closely related organisms. PMID- 17109185 TI - Authentication of traditional Chinese medicine using infrared spectroscopy: distinguishing between ginseng and its morphological fakes. AB - The quality of pharmaceutical products such as ginseng is important for ensuring consumer safety and efficacy. Ginseng is an expensive herb, and adulteration with other cheaper products may occur. Quality assurance of ginseng is needed since many of its commercial products now come in various formulations such as capsules, powder, softgels and tea. Thus traditional means of authentication via smell, taste or physical appearance are hardly reliable. Herbs like ginseng tend to exhibit characteristic infrared fingerprints due to their different chemical constituents. Here we report for the first time a rapid means of distinguishing American and Asian ginsengs from two morphological fakes--sawdust and Platycodon grandiflorum, via pattern differences and principal component analysis of their infrared spectra. Our results show that ginseng can be distinguished from both sawdust and Platycodon grandiflorum, hence there is a potential of using infrared spectroscopy as a novel analytical technique in the authentication of ginseng. PMID- 17109186 TI - HBcAg-specific CD4+CD25+ regulatory T cells modulate immune tolerance and acute exacerbation on the natural history of chronic hepatitis B virus infection. AB - Acute exacerbations (AEs) of chronic hepatitis B (CH-B) are accompanied by increased T cell responses to hepatitis B core and e antigens (HBcAg/HBeAg). Why patients are immunotolerant (IT) to the virus and why AEs occur spontaneously on the immunoactive phase remain unclear. The role of HBcAg-specific CD4(+)CD25(+) regulatory T (T(reg)) cells in AE and IT phases was investigated in this study. The SYFPEITHI scoring system was employed to predict MHC class II-restricted epitope peptides on HBcAg overlapping with HBeAg that were used for T(reg)-cell cloning and for the construction of MHC class II tetramers to measure T(reg) cell frequencies (T(reg) f). The results showed that HBcAg-specific T(reg) f declined during AE accompanied by increased HBcAg peptide-specific cytotoxic T lymphocyte frequencies. Predominant Foxp3-expressing T(reg) cell clones were generated from patients on the immune tolerance phase, while the majority of Th1 clones were obtained from patients on the immunoactive phase. T(reg) cells from liver and peripheral blood of CH-B patients express CD152 and PD1 antigens that exhibit suppression on PBMCs proliferation to HBcAg. These data suggest that HBcAg peptide-specific T(reg) cells modulate the IT phase, and that their decline may account for the spontaneous AEs on the natural history of chronic hepatitis B virus infection. PMID- 17109187 TI - Current issues in undergraduate psychiatry education: the findings of a qualitative study. AB - BACKGROUND: Recruitment into psychiatry is correlated with the quality of undergraduate medical school teaching programmes and with a commitment of major resources to teaching students. There is an extensive literature related to attitudes towards psychiatry but less on the learning and teaching of psychiatry. AIMS: To identify the current issues in undergraduate psychiatric education in the UK for lead teachers at UK medical schools. METHOD: Semi-structured telephone interviews with psychiatric leads at UK medical schools. A total of 26 participants were interviewed from 23 different medical schools. RESULTS: Three key areas of problems were identified: issues related to teaching personnel (e.g. conflict of time), teaching resources and impact of teaching on recruitment (e.g. role models; stigma). Eight potential solutions to address the problems were identified and these included improving the quality of teaching, improving the perceived value of the discipline and recruiting teachers. CONCLUSION: There are several problems facing teachers in psychiatry but the teachers are also able to identify solutions which need support from both education and health if they are to be implemented. PMID- 17109188 TI - Teachers' interactions and their collaborative reflection processes during peer meetings. AB - Teachers' reflections are often narrowly focused on technical questions ('how to') and less on the underlying moral, political and emotional aspects of their functioning. However, for a better understanding of teaching practice it is important to uncover beliefs and values that usually remain implicit. Meeting with others is considered crucial for enhancing the quality of teachers' reflections. However, little is known about how any beneficial effects of such meetings are brought about. We explored the relationship between medical teachers' interactions and collaborative reflection processes during peer meetings. Five experienced teachers and a teacher trainer participated in the study. Three peer meetings were videotaped and transcribed. Teachers' interactions and collaborative reflective processes were analysed. The interactions promoted reflection not only on technical questions but also on moral, political, and emotional issues. 'Guiding/directing', 'proposing an alternative' and 'exploring an alternative' appeared to be the principal interactions. The results may be useful for teachers and trainers who are considering organising and/or improving peer meetings. PMID- 17109189 TI - Degradation of polyethylene succinate (PES) by a new thermophilic Microbispora strain. AB - Thermophilic actinomycetes were isolated from sediment of the Chingshuei hot spring in north Taiwan, and the strain HS 45-1 was selected from colonies which formed distinct clear zones on agar plate with emulsified polyethylene succinate (PES). The film of PES disappeared within 6 days in liquid cultures at 50 degrees C. The strain HS 45-1 was also able to degrade poly (epsilon-carpolactone) (PCL) and poly (3-hydroxybutyrate) (PHB) films completely within 6 days in liquid cultures. Basing on the results of phynotypic characteristics, phylogenetic studies and DNA-DNA hybridization, strain HS 45-1 should be assigned to Micorbispora rosea subsp. taiwanensis. PMID- 17109190 TI - The use of differential item functioning analyses to identify cultural differences in responses to the EORTC QLQ-C30. AB - INTRODUCTION: The European Organisation for Research and Treatment of Cancer (EORTC) QLQ-C30 is a widely used health-related quality of life instrument. The main aim of this study is to investigate whether there are international differences in response to the questionnaire that can be explained by cultural factors. METHODS: Analyses involved a database of 106 separate studies including data from over 28,000 respondents. Differential item functioning (DIF) analyses using logistic regression were conducted for each item of the EORTC QLQ-C30 with respect to cultural/geographic group. Results were qualitatively compared with previously reported DIF analyses by translation to explore whether the source of the DIF was more likely to be linguistic or cultural in nature. RESULTS: Although most response patterns were similar, there were a number of international differences in how the questionnaire was answered. The largest variations were found in the results for Eastern Europe and East Asia. Results for the UK, the US and Australia tended to be similar. Many of the European results followed patterns that were more clearly explained when grouped by translation than when grouped by geographical region. DISCUSSION: Our results suggest that, in general, the EORTC QLQ-C30 is suitable for use in a wide variety of countries and settings. Some response variations that have the potential to affect the results of international studies were identified, but it was not always clear whether the source of the variation was primarily linguistic or cultural. PMID- 17109191 TI - Quality of life in cancer patients: The role of optimism, hopelessness, and partner support. AB - The interaction of optimism, hopelessness and social support as predictors of Health-Related Quality of Life (HRQL) among seriously ill people is not well understood. Also, the impact of partner characteristics on patient quality of life has often been overlooked. In this study the relationships between optimism, hopelessness, partner support and HRQL were investigated in 155 cancer patients and their partners. Special attention was given to the effects of optimism and hopelessness as mediators and moderators in the partner support-HRQL relationship. The impact of partner optimism and hopelessness on perceived partner support and patient HRQL was also studied. The results indicated substantial gender differences in the relationships between the study variables. High levels of partner support were associated with female patients' optimistic appraisals, and together they predicted better HRQL at 8 months follow-up. Partial support was found for the effect of optimism as a mediator. For male patients, low hopelessness was the key variable predicting good HRQL. Clear evidence for the moderator effects of optimism/hopelessness was not found, and the expected impact of partner's characteristics on partner support or patient HRQL could not be confirmed. Although partner support, patient optimism and hopelessness all appeared to be important determinants of HRQL in cancer patients, the relationships between these variables differed by gender. The proposed mediation and moderation models needs to be confirmed in future studies. PMID- 17109192 TI - Unique vascular phenotypes following over-expression of individual VEGFA isoforms from the developing lens. AB - Formation of a correctly organised vasculature and subsequently embryonic survival is critically dependent on the dosage and site-specific expression of VEGF. Murine VEGF exists in three common isoforms (viz. 120, 164 and 188 amino acids) having different organ specific distribution levels. Gene knock-in studies show that expression of any of the individual isoforms of VEGF extends survival until birth, although each is associated with distinct organ-specific abnormalities. Comparison of the effects of VEGF isoform expression is complicated by the general lethality of mis-expression, in addition to cumulative effects of adjacent tissues from the inappropriately patterned vasculature. Here we investigate the effects of over-expression of individual VEGFA isoforms from the lens-specific alphaA-Crystallin promoter and characterise their effects on the vessel morphology of the hyaloid and developing retinal vasculature. Since the hyaloid vasculature is an anatomically distinct, transient vasculature of the eye, comprising 3 cell types (endothelium, pericytes and macrophages) it is possible to more readily interpret the role of individual VEGF-A isoforms in vascular pattern formation in this model. The severity of the vascular phenotype, characterised by a hyperplastic hyaloid at E13.5 and subsequently retinal vascular patterning and ocular defects, is most severe in transgenics over expressing the more diffusible forms of VEGFA (120 and 164), whereas in VEGFA(188) transgenics the hyaloid vascular defects partially resolve post natally. The results of this study indicate that individual isoforms of VEGFA induce distinct vascular phenotypes in the eye during embryonic development and that their relative doses provide instructive cues for vascular patterning. PMID- 17109193 TI - Vascular endothelial growth factor receptor-1 (VEGFR-1/Flt-1): a dual regulator for angiogenesis. AB - Vascular endothelial growth factor receptor-1 (VEGFR-1) is a member of the VEGFR family, and binds VEGF-A, PlGF, and VEGF-B. An important feature of VEGFR-1 is that, unlike other VEGFR genes, it expresses two types of mRNA, one for a full length receptor and another for a soluble short protein known as soluble VEGFR-1 (sFlt-1). The binding-affinity of VEGFR-1 for VEGF-A is one order of magnitude higher than that of VEGFR-2, whereas the kinase activity of VEGFR-1 is about 10 fold weaker than that of VEGFR-2. Through its ligand-binding region and by trapping ligands, VEGFR-1 plays a negative role in angiogenesis at embryogenesis. In adulthood, however, VEGFR-1 is expressed not only on endothelial cells but also on macrophages, and promotes the function of macrophages, inflammatory diseases, cancer metastasis, and atherosclerosis via its kinase activity. Soluble VEGFR-1 is abnormally overexpressed in the placenta of preeclamptic patients, and suggested to cause the major pathological symptoms on the maternal side such as hypertension and renal dysfunction, most likely by blocking the physiological VEGF-A. VEGFR-1 including its soluble form is involved in a variety of human illnesses, making it an important target in the development of new strategies to suppress disease. PMID- 17109194 TI - Three-dimensional (3D) reconstruction and quantitative analysis of the microvasculature in medulloblastoma and ependymoma subtypes. AB - In the World Health Organisation (WHO) classification of tumours of the nervous system, four main histopathological subtypes of medulloblastomas (classic medulloblastoma, desmoplastic medulloblastoma, medulloblastoma with extensive nodularity and advanced neuronal differentiation and large cell/anaplastic medulloblastoma) as well as of ependymal tumours (low-grade ependymoma, anaplastic ependymoma, myxopapillary ependymoma and subependymoma) are recognised. Under the hypothesis that the microvascular architecture of tumours is a reflection of the histopathological subtype, we performed three-dimensional reconstructions of the microvasculature in these subtypes of medulloblastomas and ependymal tumours using computerised image analysis. In addition, we quantitatively assessed three microvascular parameters (number, area, perimeter) in these neoplasms. Three-dimensional reconstructions showed a dense pattern of irregular vessels in classic and large cell medulloblastoma. In desmoplastic medulloblastoma and medulloblastoma with extensive nodularity, the vessels were more unevenly distributed and organised around the nodular areas. Classic medulloblastoma and large cell medulloblastoma had on average the largest vessel area and perimeter. The highest number of vessels was seen in classic medulloblastoma and medulloblastoma with extensive nodularity. Three-dimensional analysis of ependymal tumours showed that low-grade ependymoma had larger but fewer vessels compared to anaplastic ependymoma, while myxopapillary ependymoma had a complex, heterogeneous pattern of vessels and subependymoma few but regular vessels. In ependymal tumours, the highest values for vessel number, vessel area and vessel perimeter were found in anaplastic ependymoma and the lowest values in subependymoma. We conclude that our three-dimensional reconstructions shed unprecedented light on the tumour vasculature in medulloblastomas and ependymal tumours and expect that such reconstructions are helpful tools for further studies on tumour angiogenesis. PMID- 17109195 TI - c7E3 Fab inhibits human tumor angiogenesis in a SCID mouse human skin xenograft model. AB - The alphavbeta3 integrin plays an important role in tumor growth and angiogenesis. Inhibition of this receptor by intact bivalent antibodies has been shown to inhibit angiogenesis and tumor growth. In this study we tested the chimeric Fab of 7E3 (c7E3 Fab), an antibody reactive with human platelet GPIIb/IIIa and alphavbeta3 to determine if it would inhibit in vivo angiogenesis and tumor growth in a SCID mouse/human skin tumor growth and angiogenesis model. c7E3 Fab inhibited human tumor angiogenesis and tumor growth. These data suggest monovalent antibody fragments devoid of antibody effector function can have efficacy in preclinical models of angiogenesis. PMID- 17109196 TI - Immunohistochemical measurement of endothelial cell apoptosis and proliferation in formalin-fixed, paraffin-embedded human cancer tissue. AB - There is a need for simple, reproducible methodology for measurement of endothelial cell (EC) apoptosis and proliferation in formalin-fixed, paraffin embedded (FFPE) human tissue obtained in clinical trials of potential anti angiogenic agents. Therefore, we developed colorimetric, dual-label immunohistochemical techniques for use on FFPE tissue, based on the use of single stranded (ss) DNA and Ki-67 as markers of EC apoptosis and proliferation, respectively. Digital image analysis was used to obtain the total tumour microvessel density (MVD), from which the EC apoptosis index (AI) and proliferation index (PI) were derived manually as the number of positive ECs per vessel. Immunohistochemical measurement of EC apoptosis and proliferation was validated on human colorectal cancer liver metastases from a randomized, placebo controlled trial of the selective cyclooxygenase-2 inhibitor rofecoxib. Proliferating and apoptotic ECs clustered in discrete areas of the tumour vasculature. ECAI (median [inter-quartile range, IQR] 0.0018 [0.0003-0.0064]) and ECPI (median [IQR] 0.0043 [0.002-0.014]) values were low and highly variable in tumours from both placebo- and rofecoxib-treated patients. Our novel dual-label immunohistochemical methods will be generally applicable in FFPE human cancer tissue and should prove invaluable for measuring the anti-angiogenic activity of experimental therapies in clinical trials. The low absolute level of and variability in EC apoptosis and proliferation detectable in human colorectal cancer liver metastases indicates that similar intervention studies using these end-points will need to ensure adequate size in order to be able to detect anti angiogenic activity by immunohistochemical methods. PMID- 17109197 TI - Inhibition of laminin-8 in vivo using a novel poly(malic acid)-based carrier reduces glioma angiogenesis. AB - We have previously shown that laminin-8, a vascular basement membrane component, was overexpressed in human glioblastomas multiforme and their adjacent tissues compared to normal brain. Increased laminin-8 correlated with shorter glioblastoma recurrence time and poor patient survival making it a potential marker for glioblastoma diagnostics and prediction of disease outcome. However, laminin-8 therapeutic potential was unknown because the technology of blocking the expression of multi-chain complex proteins was not yet developed. To inhibit the expression of laminin-8 constituents in glioblastoma in vitro and in vivo, we used Polycefin, a bioconjugate drug delivery system based on slime-mold Physarum polycephalum-derived poly(malic acid). It carries an attached transferrin receptor antibody to target tumor cells and to deliver two conjugated morpholino antisense oligonucleotides against laminin-8 alpha4 and beta1 chains. Polycefin efficiently inhibited the expression of both laminin-8 chains by cultured glioblastoma cells. Intracranial Polycefin treatment of human U87MG glioblastoma bearing nude rats reduced incorporation of both tumor-derived laminin-8 chains into vascular basement membranes. Polycefin was thus able to simultaneously inhibit the expression of two different chains of a complex protein. The treatment also significantly reduced tumor microvessel density (p < 0.001) and area (p < 0.001) and increased animal survival (p < 0.0004). These data suggest that laminin-8 may be important for glioblastoma angiogenesis. Polycefin, a versatile nanoscale drug delivery system, was suitable for in vivo delivery of two antisense oligonucleotides to brain tumor cells causing a reduction of glioblastoma angiogenesis and an increase of animal survival. This system may hold promise for future clinical applications. PMID- 17109198 TI - Image analysis measurements of the microvascularisation in endometrium, superficial and deep endometriotic tissues. AB - The aim of this study was to evaluate precisely the microvascularisation of endometrium, superficial and deep endometriotic lesions, in progestin-treated and non-treated patients suffering from endometriosis. METHODS: A population of 66 women was constituted. Immunohistochemistry was carried out with a specific marker of the endothelial cells (CD31). The number of vessels and the vessel area were assessed by a computer image analysis system. RESULTS: The number of vessels per mm2 were 211, 216, 225 and the vessel area was 270, 141 and 194 microm2, respectively in endometria, superficial and deep endometriotic lesions of untreated women. In endometria, superficial and deep endometriotic lesions of progestin-treated women the number of vessels were respectively 129, 149, and 181 per mm2 and the vessel area was 369, 474 and 254 microm2. CONCLUSION: Statistically significant data indicate that endometriotic lesions are heterogeneous and suggest that progestin treatment induces a reduction in number and a concomitant dilation of microvessels with more microvascular changes in endometrium and superficial endometriotic lesions than in deep endometriotic lesions. PMID- 17109200 TI - Cardioverter defibrillator implantation in a patient with double chambered right ventricle. AB - Double-chambered right ventricle (DCRV) is a rare form of congenital heart disease in which the right ventricle is divided into a high-pressure inlet site and a low-pressure outlet site by anomalous muscle bands. We describe a patient with DCRV presenting with ventricular tachycardia. PMID- 17109199 TI - Experimental evaluation of the detectability of submillimeter atherosclerotic lesions in ex vivo human iliac arteries with ultrahigh-field (7.0 T) magnetic resonance imaging. AB - BACKGROUND: To evaluate the ability of ultrahigh-field magnetic resonance imaging (MRI) to accurately depict the composition of the human arterial vessel wall ex vivo and to detect early atherosclerotic lesion formation in comparison to histology. METHODS: Eight iliac artery specimens with low-grade atherosclerotic lesions obtained from human organ donors were studied. Three-dimensional, high resolution MRI (spatial resolution: 79 x 79 x 109 microm) was performed using T1 , T2- and proton density (PD)-weightings (7.0 Tesla MR system, Bruker Pharmascan). A total of 36 MR slices and corresponding histological sections were matched for comparative evaluation of area measurements of lumen, media and adventitia and--if present--plaque size. Statistical correlation between histology and MR measurements was tested and a ROC-analysis was performed to determine the plaque size being predictive of correctly identifying atherosclerotic lesions with MRI. RESULTS: The areas of vessel lumen and media as measured on T1-, T2- and PD-weighted MR images showed a strong correlation with the corresponding histological measurements (r = 0.84 to r = 0.89; P < 0.01), however, a systematic overestimation of 34-41% was found. For the area of adventitia, only a moderate, though significant, correlation (r = 0.55 to r = 0.62; P < 0.01) could be demonstrated with a similar overestimation by MRI (38 43%). With T1-weighted MRI, sensitivity and specificity for the detection of plaques > 4.0 mm(2) were 79% and 91%, respectively. With T2- and PD-weighted MRI, however, sensitivity and specificity for the detection of plaques > 0.4 mm2 were 93% and 89%. CONCLUSIONS: In an experimental ex vivo setting, ultrahigh-field MRI of the human arterial vessel wall resulted in an accurate visualization of vessel wall composition when compared to histology and, thus, allowed for a quantitative assessment. T2- and PD-weighted MRI proved capable of reliably detecting submillimeter atherosclerotic lesions. PMID- 17109201 TI - Safety and feasibility of atropine added in patients with sub-maximal heart rate during exercise myocardial perfusion SPECT. AB - BACKGROUND: Failure to reach 80% of maximal predicted heart rate (HR) during exercise may render a myocardial perfusion single photon emission computed tomography (SPECT) study non-diagnostic for ischemia detection. We sought to investigate the injection of atropine in patients who fail to achieve 80% of age predicted HR during exercise performed for myocardial perfusion SPECT (MPS), defining its safety and efficacy to raise HR to adequate levels as well as its effect on MPS interpretation. METHODS AND RESULTS: Between January 2002 and December 2004, we studied 3,150 consecutive patients (2,253 men and 897 women, mean age 55 +/- 6 years) who were referred to a single office-based nuclear cardiology laboratory for MPS using SPECT imaging. One milligram of atropine was administered to patients that were unable to continue because of fatigue before reaching minimal HR, without an ischemic response (group A, n = 397). The scintigraphic results for group A were compared with those of patients who spontaneously achieved target HR (group B, n = 2,753). In group A, mean HR before atropine injection was 119.5 +/- 13.6 beats per minute (bpm), and it increased up to 137.3 +/- 13.5 bpm after drug administration, with an incremental of 17.8 +/- 6.9 bpm (P < 0.0001). The mean percentage of age-related HR achieved in this group was 83.5 +/- 8.1%. In 302 of this patients (76.1%) more than 80% of their aged-related HR (86.9 +/- 5.1%) was attained. No major adverse effects occurred. When groups A and B were compared, baseline and peak HR, rate pressure product, and maximal metabolic equivalents achieved were higher in group B. There were no significant differences in the percentage of total positive perfusion studies between both groups: 210/397 patients (52.9%) in group A and 1,342/2,753 patients (48.7%) in group B (P = 0.39). Ischemia or ischemia plus scar was found in 112/397 patients (28.2%) in group A and in 923/2,753 patients (33.5%) of group B (P = 0.14). CONCLUSION: Atropine added to exercise stress testing in patients who cannot achieve their 80% age-related HR is a safe, well-tolerated, and feasible method for MPS. PMID- 17109202 TI - Primary uretero-iliac fistula: the unusual source of haematuria. AB - Uretero-iliac fistula is a rare cause of frank haematuria. The aetiology of such fistula is commonly iatrogenic. We present a unique case of a primary aorto-iliac fistula in the absence of an aneurysm or arteriovenous malformation. The diagnosis was demonstrated by ureteroscopy and real-time retrograde ureterogram. Multiple arterial embolisation of the fistula had failed, and the patient underwent a successful ureterolysis and ligation of fistula. We demonstrate the diagnostic difficulties and treatment dilemma of such rare cause of haematuria. PMID- 17109203 TI - Transpupillary thermotherapy for myopic choroidal neovascularization: 1-year follow-up: TTT for myopic CNV. AB - PURPOSE: To evaluate the use of transpupillary thermotherapy (TTT) for myopic choroidal neovascularization (CNV) retrospectively in a case series. METHODS: Ten consecutive patients with CNV secondary to high myopia were treated with TTT and followed-up for 18.3 +/- 7.1 months (range: 12-30 months). An activity scoring (AS) scheme based on CNV size, central foveal thickness as determined by optical coherence tomography (OCT), amount of subretinal hemorrhage, and fluorescein angiography (FA) staining characteristics was developed to assess the response to treatment. The main outcome measures were AS and visual acuity (VA). RESULTS: There was no significant loss or gain in VA, and the lesions stabilized in all eyes. FA revealed late staining typical of CNV scarring, and the amount of central foveal thickness decreased in all eyes, as determined by OCT. The mean AS at 3, 6, and 12 months and at the final visit were significantly lower than the preoperative AS (P<0.05). No complication occurred during or after treatment. CONCLUSION: A scheme for evaluating overall CNV activity may be useful tool for assessing the response of patients to TTT. TTT seems to stabilize myopic CNVs both clinically and as revealed by angiography, with a significant decrease in the activity of lesions. Longer term results are required to evaluate the stabilizing effect of TTT in myopic CNV. PMID- 17109204 TI - A new index of the autonomic nervous balance and its clinical usefulness; a power law 1/f like scale of a spectral analysis of the heart rate variability. AB - OBJECTIVE: This study was undertaken to develop a new autonomic scoring scale distinct from the conventional sympathetic or parasympathetic parameters cardiac autonomic response. METHODS: The parameters were obtained from a log-log scale of frequency (f ) vs. power spectral (P) amplitude of heart rate variability (HRV). A gradient value in which the graph decreases towards a high frequencies on the X axis, while showing an absolute value beta itself based on the formula 'P = f ( beta)' was newly named the 'Balance index' by implying a balanced state in the living body. The index was categorized into four bands based on the appearance of the whole frequency ranges (T-Balance Index). Therefore, we compared the responses of the following groups: the patient test group (Group PT), the normal test group (Group NT), and the normal non-test group (group NN). RESULTS: Significant differences in parameters were observed among the three groups. In addition, the difference between the 'High-side Balance Index' and the 'Low-side Balance Index' was 0.75 +/- 0.12 vs. 0.07 +/- 0.04 (p < 0.005) for groups NT vs. PT. This novel analyzing method was useful for evaluating the subtle changes in a living body to regulate the living-power (strength of vitality). Moreover, the difference between the 'T-Balance Index' and 'SV-Balance Index' was 0.18 +/- 0.12 vs. -0.21 +/- 0.15 (p < 0.01) for groups NT vs. PT. CONCLUSIONS: One application of new parameters 'Balance Index' is its ability to evaluate the vital or emotional functions of unconscious patients in a critical condition undergoing a sense challenge test. This method assists in improving our ability to measure the early stage of conscious recovery with greater accuracy by using our novel analysis method for performing senses challenge test, with the aid of media such as music. PMID- 17109205 TI - Mining the challenges of CBPR for improvements in urban health. PMID- 17109206 TI - Exploring barriers to 'respondent driven sampling' in sex worker and drug injecting sex worker populations in Eastern Europe. AB - Respondent driven sampling (RDS) has been used in several counties to sample injecting drug users, sex workers (SWs) and men who have sex with men and as a means of collecting behavioural and biological health data. We report on the use of RDS in three separate studies conducted among SWs between 2004 and 2005 in the Russian Federation, Serbia, and Montenegro. Findings suggest that there are limitations associated with the use of RDS in SW populations in these regions. Findings highlight three main factors that merit further investigation as a means of assessing the feasibility and appropriateness of RDS in this high risk population: the network characteristics of SWs; the appropriate level of participant incentives; and lack of service contact. The highly controlled and hidden nature of SW organizations and weak SW social networks in the region can combine to undermine assumptions underpinning the feasibility of RDS approaches and potentially severely limit recruitment. We discuss the implications of these findings for recruitment and the use of monetary and non-monetary incentives in future RDS studies of SW populations in Eastern Europe. PMID- 17109207 TI - Feasibility of the transseptal approach for fast and unstable left ventricular tachycardia mapping and ablation with a non-contact mapping system. AB - BACKGROUND: Radiofrequency ablation of fast and unstable left ventricular tachycardia (VT) usually requires non-contact mapping. The procedure is usually performed by a retrograde-transaortic route, requiring a double femoral artery puncture, for the 9F multielectrode catheter and the 7F ablation catheter which are advanced through the aorta and aortic valve into the left ventricle (LV). Reported limitations of the procedure are due to the stiffness of the balloon catheter, particularly in patients with tortuous peripheral arteries, atherosclerotic aorta, or with aortic stenosis. The aim of our study was to test the feasibility and assess the safety of a transseptal approach for left VT non contact mapping and ablation. MATERIALS AND METHODS: Ten patients with multiple cardiac defibrillator shocks because of fast and unstable VT were selected for non-contact mapping and ablation. After a double transseptal puncture the multielectrode catheter (Ensite Array, St. Jude Medical) was advanced through a standard 10F introducer to a stable position in the LV apex over a 260 cm length 0.035 J-tip guidewire. The ablation catheter (Celsius Thermo-cool, Biosense Webster) was then inserted through the second 8F introducer. Twenty-five monomorphic sustained ventricular tachycardia were induced and ablated at the level of the diastolic pathway or exit point revealed by unipolar isopotential mapping. The total procedural and fluoroscopy times were 209 +/- 32 min and 28.5 +/- 9.27 min, respectively, which were comparable to those described with the traditional retrograde-transaortic approach. No major complication related with the transseptal approach were reported. CONCLUSION: A transseptal approach can be a feasible and effective alternative approach for mapping and ablation of fast and unstable left VT with a non-contact mapping system. PMID- 17109208 TI - Successful ablation of a concealed parahisian accessory pathway using a remote magnetic navigation system following failure by conventional methods. AB - OBJECTIVE: Using conventional catheters, ablation of concealed parahisian accessory pathways may be difficult and high risk for heart block. METHODS AND RESULTS: We describe the case of a concealed parahisian accessory pathway with three prior attempts to ablate using conventional methods (RF and cryotherapy). Using a remote magnetic navigation system, successful ablation occurred following a single RF lesion (total fluoroscopy time 17 min). In contrast to previous attempts, the patient remained asymptomatic during follow-up (12 months). CONCLUSIONS: This is the first report of successful remote magnetic catheter ablation of a concealed parahisian AP. Magnetic catheter stability during RF application likely contributed to the success of this procedure and may have minimized the risk of AV block. PMID- 17109209 TI - Rational design of a dual-mode optical and chemical prodrug. AB - PURPOSE: The purpose of this study is to demonstrate the rational design and behaviour of the first dual-mode optical and chemical prodrug, exemplified by an acetyl salicylic acid-based system. METHODS: A cyclic 1,4-benzodioxinone prodrug was synthesised by reaction of 3,5-dimethoxybenzoin and acetyl salicoyl chloride with pyridine. After purification by column chromatography and recrystallization, characterization was achieved using infrared and NMR spectroscopies, mass spectrometry, elemental analysis and single crystal X-ray diffraction. Light triggered drug liberation was characterised via UV-visible spectroscopy following low-power 365 nm irradiation for controlled times. Chemical drug liberation was characterised via UV-visible spectroscopy in pH 5.5 solution. RESULTS: The synthetic method yielded pure prodrug, with full supporting characterisation. Light-triggered drug liberation proceeded at a rate of 8.30x10(-2) s-1, while chemical, hydrolytic liberation proceeded independently at 1.89x10(-3) s-1. The photochemical and hydrolytic reactions were both quantitative. CONCLUSIONS: This study demonstrates the first rational dual-mode optical and chemical prodrug, using acetyl salicylic acid as a model, acting as a paradigm for future dual-mode systems. Photochemical drug liberation proceeds 44 times faster than chemical liberation, suggesting potential use in drug-eluting medical devices where an additional burst of drug is required at the onset of infection. PMID- 17109210 TI - Antiangiogenic effect of bile acid acylated heparin derivative. AB - PURPOSE: Chemically modified heparin-DOCA was prepared and found to have markedly lower anticoagulant activity than heparin. In the present study, we elucidated the antiangiogenic and antitumoral activities of heparin-DOCA derivative. METHODS: To evaluate the antiangiogenic and antitumoral effects of heparin-DOCA, capillary-like tube formation assay, Matrigel plug assay in vivo, western blotting for FGFR phosphorylation, ERK and p38 MAPK activities, tumor growth of SCC in vivo and immunostaining of blood vessels in tumor tissues were performed. RESULTS: Heparin-DOCA inhibited capillary-like tubular structures of endothelial cells and bFGF-induced neovascularizations in Matrigel plug assays. Signaling experiments showed that heparin-DOCA significantly inhibited angiogenesis by suppressing the phosphorylation of FGFR and its downstream signal pathways (ERK and p38 MAPK activities). The antiangiogenic activity of this heparin derivative was found to be closely associated with antitumoral activity in a mouse model. In addition, histological evaluations supported the inhibitory effect of heparin DOCA on blood vessel formation in tumor tissues. CONCLUSION: Heparin-DOCA derivative exerted a significant antitumoral effect by inhibiting angiogenesis resulting from the disruption of FGF/FGFR and its downstream signal pathways, and could be applied to treat various angiogenic diseases. PMID- 17109212 TI - Sorbitol crystallization can lead to protein aggregation in frozen protein formulations. AB - PURPOSE: This work examines the cause of aggregation of an Fc-fusion protein formulated in sorbitol upon frozen storage for extended periods of time at -30 degrees C. MATERIALS AND METHODS: We designed sub-ambient differential scanning calorimetry (DSC) experiments to capture the effects of long-term frozen storage. The physical stability of formulation samples was monitored by size exclusion high performance liquid chromatography (SE-HPLC). RESULTS: DSC analysis of non frozen samples shows the expected glass transitions (Tg') at -45 degrees C for samples in sorbitol and at -32 degrees C in sucrose. In time course studies where sorbitol formulations were stored at -30 degrees C and analyzed by DSC without thawing, two endothermic transitions were observed: a melting endotherm at -20 degrees C dissipated over time, and a second endotherm at -8 degrees C was seen after approximately 2 weeks and persisted in all later time points. Protein aggregation was only seen in the samples formulated in sorbitol and stored at -30 degrees C, correlating aggregation with the aforementioned melts. CONCLUSIONS: The observed melts are characteristic of crystalline substances and suggest that the sorbitol crystallizes over time. During freezing, the excipient must remain in the same phase as the protein to ensure protein stability. By crystallizing, the sorbitol is phase-separated from the protein, which leads to protein aggregation. PMID- 17109211 TI - Micellar nanocarriers: pharmaceutical perspectives. AB - Micelles, self-assembling nanosized colloidal particles with a hydrophobic core and hydrophilic shell are currently successfully used as pharmaceutical carriers for water-insoluble drugs and demonstrate a series of attractive properties as drug carriers. Among the micelle-forming compounds, amphiphilic copolymers, i.e., polymers consisting of hydrophobic block and hydrophilic block, are gaining an increasing attention. Polymeric micelles possess high stability both in vitro and in vivo and good biocompatibility, and can solubilize a broad variety of poorly soluble pharmaceuticals many of these drug-loaded micelles are currently at different stages of preclinical and clinical trials. Among polymeric micelles, a special group is formed by lipid-core micelles, i.e., micelles formed by conjugates of soluble copolymers with lipids (such as polyethylene glycol phosphatidyl ethanolamine conjugate, PEG-PE). Polymeric micelles, including lipid core micelles, carrying various reporter (contrast) groups may become the imaging agents of choice in different imaging modalities. All these micelles can also be used as targeted drug delivery systems. The targeting can be achieved via the enhanced permeability and retention (EPR) effect (into the areas with the compromised vasculature), by making micelles of stimuli-responsive amphiphilic block-copolymers, or by attaching specific targeting ligand molecules to the micelle surface. Immunomicelles prepared by coupling monoclonal antibody molecules to p-nitrophenylcarbonyl groups on the water-exposed termini of the micelle corona-forming blocks demonstrate high binding specificity and targetability. This review will discuss some recent trends in using micelles as pharmaceutical carriers. PMID- 17109213 TI - Aqueous-soluble, non-reversible lipid conjugate of salmon calcitonin: synthesis, characterization and in vivo activity. AB - PURPOSE: A novel, non-reversible, aqueous-based lipidization strategy with palmitic acid as a model lipid was evaluated for conjugation with salmon calcitonin (sCT). MATERIALS AND METHODS: A water-soluble epsilon-maleimido lysine derivative of palmitic acid was synthesized from reaction of palmitic acid N succinimidyl ester and epsilon-maleimido lysine. The latter was generated from reaction of alpha-Boc-lysine and methylpyrrolecarboxylate, with subsequent deprotection of the Boc group. The palmitic derivative was further conjugated with sCT via a thio-ether bond to produce Mal-sCT in aqueous solution. The identity and purity of Mal-sCT was confirmed by Electrospray Ionisation Mass spectrometry (ESI-MS) and HPLC. RESULTS: Yield of Mal-sCT was 83%. Dynamic light scattering and circular dichroism data suggested that Mal-sCT presented as a stable helical structure in aqueous solutions of varying polarity, with a propensity to aggregate at concentrations above 11 microM. Cellular uptake of Mal sCT was twice that of sCT in the Caco-2 cell model, and the conjugate was more resistant to liver enzyme degradation. Mal-sCT exhibited comparable hypocalcemic activity to sCT when administered subcutaneously in the rat model at sCT equivalent dose of 0.114 mg/kg. Peroral Mal-sCT, however, produced variability in therapeutic outcome. While four out of six rats did not respond following intragastric gavage with Mal-sCT, two rats showed significantly suppressed plasma calcium levels (approximately 60% of baseline) for up to 10 h. CONCLUSION: A novel non-reversible, water-soluble lipid conjugate of sCT was successfully synthesized that showed (1) different aggregation behavior and secondary structure, (2) improved enzymatic stability and cellular uptake, and (3) comparable hypocalcemic activity in vivo compared to sCT. PMID- 17109214 TI - Involvement of UDP-glucuronosyltransferases in the extensive liver and intestinal first-pass metabolism of flavonoid baicalein. AB - PURPOSE: The present study aims to investigate the involvement of UDP glucuronosyltranferase (UGT) in the extensive liver and intestinal first-pass glucuronidation of baicalein (B) in both rats and humans and also to study sulfation and P450 mediated hydroxylation of B. MATERIALS AND METHODS: B was incubated with liver and intestine microsome, cytosol, S9 fractions from human, rat and various human recombinant UGT isozymes, respectively. The generated metabolites were identified by HPLC/MS/MS and quantified by HPLC/UV. RESULTS: Three metabolites of B namely baicalein 7-O-glucuronide (BG), the isomer of baicalein 7-O-glucuronide (BG'), and baicalein sulfate were found. BG, the predominant metabolite of B, was extensively generated in liver and jejunum microsomes in both humans and rats. Its formation was mainly catalyzed by UGT 1A9 and also mediated by UGT 1A1, 1A3, 1A8, 1A7 and 2B15 with different kinetic profiles. UGT 1A8 mediated formation of BG' was mainly found in human intestine and rat liver microsomes. Sulfation and P450 mediated hydroxylation of B were much less significant than glucuronidation. CONCLUSIONS: Extensive liver and intestinal first-pass glucuronidation of B were found in both humans and rats. Under the current experimental conditions, UGT 1A9 and UGT 1A8 demonstrated the fastest formation rate of BG in human liver preparations and BG' in human intestine preparations, respectively. PMID- 17109215 TI - Design for optimized topical delivery: Prodrugs and a paradigm change. AB - In theory, topical delivery has substantial potential to treat local and some systemic disease states more effectively than systemic delivery. Unfortunately many, if not most, drug candidates for topical delivery lack the requisite physicochemical properties that would allow them to permeate the skin to a clinically useful extent. One way to overcome this obstacle to effective topical delivery is to make a transient derivative of the drug, a prodrug, with the correct physicochemical properties. But what are those correct properties and can the directives for the design of prodrugs be applied to the design of new drugs, their analogs or homologs? For some time increasing the lipid solubility (S (LIPID)) or its surrogate, the partition coefficient between a lipid (LIPID) and water (AQ) (K (LIPID:AQ)), has been the standard working paradigm for increasing permeation of the skin, and the permeability coefficient (P = distance/time) has been the quantitative measure of the result. However, even the earliest reports on non-prodrugs such as alcohols showed that working paradigm was incorrect and that P should not be the relevant measure of permeation. The shorter chain and more water soluble alcohols exhibiting lower K (LIPID:AQ) values gave the greater flux values (J = amount/area x time; the more clinically relevant measure of permeation), regardless of whether they were applied neat or in an aqueous vehicle, while P showed opposite trends for the two applications. Subsequently a large volume of work has shown that, for prodrugs and non-prodrug homologs or analogs alike, S (AQ) (not solubility in the vehicle, S (VEH)) as well as S (LIPID) should be optimized to give maximum flux from any vehicle, J (MVEH): a new working paradigm. The dependence of J (MVEH) on S (AQ) is independent of the vehicle so that S (AQ) as well as S (LIPID) are descriptors of the solubilizing capacity of the skin or S (M1) in Fick's law. The inverse dependence of J (or P) on molecular weight (MW) or volume (MV) remains. Here we review the literature that leads to the conclusion that a new working paradigm is necessary to explain the experimental data, and argue for its use in the design of new prodrugs or in the selection of candidate analogs or homologs for commercialization. PMID- 17109216 TI - Endoscopic Doppler ultrasound versus endoscopic stigmata-directed management of acute peptic ulcer hemorrhage: a multimodel cost analysis. AB - Recurrent bleeding from acute peptic ulcer hemorrhage is problematic. Studies have shown that Doppler ultrasound (DOP-US) is useful in decreasing rebleeding. We analyzed associated costs and outcomes to better define the role of DOP-US versus Conventional (Forrest classification endoscopic stigmata) in the management of acute peptic ulcer bleeding. Two separate decision analyses were constructed. Recurrent bleeding, failed esophagogastroduodenoscopy (EGD) hemostasis, complications, and surgery rates were derived from medical literature. Costs were based on Medicare data. DOP-US is preferred over Conventional in acute peptic ulcer bleeding with average cost savings per patient ranging from 853 dollars (decision-tree modeling) to 1,160 dollars (Monte Carlo simulation). High-dose intravenous proton-pump inhibitors lowered rates of recurrent bleeding for both Conventional and DOP-US, resulting in a lower but still persistent average cost savings per patient for DOP-US (decision-tree modeling = 328 dollars, Monte Carlo simulation = 560 dollars). This decision analyses identified DOP-US as the preferred cost-minimizing strategy in acute peptic ulcer hemorrhage. Results of cost analyses were most dependent on hospitalization costs and recurrent bleeding rates. PMID- 17109217 TI - Physical activity behavior, motivational readiness and self-efficacy among Ontarians with cardiovascular disease and diabetes. AB - This cross-sectional study examined physical activity and its correlates among 355 diabetes, 144 cardiovascular disease, 75 diabetes and cardiovascular disease, and 390 residents with cardiovascular risk factors. Community residents (N=2566) were screened by telephone, and 964 participants completed a self-report survey. Non-diabetes participants participated in a greater range of physical activities (p<.001), more frequently (p=.013). Diabetes participants had lower physical activity readiness and efficacy (ps<.009). In a regression model (p<.001), region and disease, work, marital and smoking status were significant correlates of physical activity frequency. Interventions which increase motivational readiness and efficacy among diabetics are required to prevent and delay complications, particularly in regions with environmental barriers such as cold weather and homogeneous, low-density land use. PMID- 17109218 TI - Genetic diversity and population structure of yellow camellia (Camellia nitidissima) in China as revealed by RAPD and AFLP markers. AB - Camellia nitidissima, a rare plant but a useful genetic resource for commercial cultivation of ornamental camellias, is distributed in a narrow region of South China and North Vietnam. In this study, RAPD and AFLP markers were used to assess the genetic diversity and population structure of six natural populations of C. nitidissima from Guangxi in South China. Twenty RAPD primers amplified 183 bands, of which 143 bands were polymorphic, and 8 AFLP primer pairs produced 502 bands, of which 364 were polymorphic. Independent as well as combined analyses of the cluster analyses of the RAPD and AFLP fragments showed that the six populations could be classified into two major genetic groups corresponding to the Nanning and Fangcheng areas. The Mantel test revealed significant correlation between the genetic and geographic distances of C. nitidissima populations (r = 0.953, p = 0.036). AMOVA analysis allowed the partitioning of the genetic variation between groups (36.09%), among populations within groups (25.78%), and within populations (38.14%). An understanding of both the genetic diversity and the population structure of C. nitidissima in China can also provide insight into the conservation and management of this endangered species. PMID- 17109219 TI - Genetic variability in Hesperozygis ringens Benth. (Lamiaceae), an endangered aromatic and medicinal plant of southern Brazil. AB - Hesperozygis ringens Benth. (Lamiaceae) is an aromatic and medicinal plant, characterized by a high concentration of isopulegone; it is endemic to the mountains of southeast Rio Grande do Sul, Brazil, and currently is considered endangered. The present study evaluated the intra- and interpopulation genetic variability of H. ringens by means of RAPD and ISSR molecular markers. The results showed that H. ringens populations are genetically structured, with low gene flow between populations, confirming the fragmentation imposed by anthropic action. Populations from the two areas of occurrence are genetically different. Low intrapopulation variability and heterozygosity were detected, indicating genetic drift and inbreeding. Based on the data, conservation strategies are discussed. PMID- 17109220 TI - Suicide probability scale and its utility with adolescent psychiatric patients. AB - This study examined the factor structure, reliability, and validity of the Suicide Probability Scale (SPS) in a sample of 226 (80 male, 146 female) adolescent psychiatric inpatients. Confirmatory factor analyses provided only some support for the original subscales. Exploratory factor analyses revealed some overlap with the original scales, but the factors differed by gender. Internal consistency of the original factors was somewhat better than the factors derived from this sample. A negative change in SPS scores over the course of the inpatient admission was related to risk for readmission due to repeat suicidal behavior. PMID- 17109221 TI - Reliability and validity of the Chinese version of the multidimensional anxiety scale for children among Chinese secondary school students. AB - The objective of the current study was to develop a Chinese translation of the Multidimensional Anxiety Scale for Children (MASC) [March (1997) Multidimensional anxiety scale for children: Technical manual, Multi health systems, Toronto, ON], and to evaluate its reliability and validity. The original version of the MASC was translated into Chinese (MASC-C) and administered to 1,538 Chinese adolescents between the ages of 14 and 19. In comparison to the American normative sample [March (1997) Multidimensional anxiety scale for children: Technical manual, Multi health systems, Toronto, ON], Chinese adolescents reported significantly higher scores on the subscales of social anxiety and separation anxiety. Girls reported higher levels of anxiety on all subscales than males. Participants between the ages of 16 and 19 reported higher scores on the physical symptoms and harm avoidance subscales. The MASC-C exhibited strong internal consistency (Cronbach's alpha coefficient was 0.91 and the mean inter item correlation coefficient was 0.20) and moderate test-retest reliability (intra-class correlation coefficient was 0.84 over a one-month interval). MASC scores inter-correlated a small to moderate degree with measures assessing negative life events and depressive symptoms indicating acceptable convergent validity. The results of confirmatory factor analyses indicated that the four factor structure of the MASC was suitable for the Chinese sample. The four factor structure was also invariant across sex and age. As the Chinese translation of the MASC indicated high levels of reliability and validity, the MASC-C is appropriate for assessing anxiety in Chinese adolescents. PMID- 17109222 TI - A brief screening measure of adolescent risk behavior. AB - This study examined the factor structure and reliability of a brief but comprehensive measure, the adolescent risk inventory (ARI), designed to assess adolescent risk behaviors and attitudes. Measures assessing demographics and risk behaviors were administered to 134 youth (ages 12-19) in psychiatric treatment. A confirmatory factor analysis of the four attitude scales (HIV Anxiety, HIV Prevention Self-Efficacy, General Distress, and General Risk) revealed excellent goodness of fit statistics. Exploratory factor analysis of the behavior items revealed three behavior factors (Sex Risk, Abuse/Self-Harm, and Acting Out). The preliminary analysis suggested that all subscales had reasonable internal consistency reliability and appeared to be independent measures, rather than part of a single unitary construct. Differences emerged based on gender, sexual activity status, and trauma history. Exploratory regression analyses revealed that, even when controlling for demographic factors and sex risk attitudes (e.g., HIV Prevention Self-Efficacy), Abuse/self-harm behaviors were highly significantly predictive of sex risk. These analyses suggest that the ARI can be useful in quickly identifying the broad range of risk behaviors found among adolescents with psychiatric disorders. PMID- 17109223 TI - Delivering a very low birth weight infant and the subsequent risk of divorce or separation. AB - BACKGROUND: The simultaneous rise over the last two decades in the U.S. in the proportion of VLBW (<1500 grams) deliveries and the improvement in their chance of survival has increased the number of families caring for VLBW infants and children. The families of VLBW infants with adverse outcomes can face psychological and monetary stresses, which in turn may influence marital instability and increase the risk of divorce or separation. The purpose of this paper is to identify the relationship of having a VLBW birth with the probability of divorce or separation in the first two years following delivery. METHODS: We use data from the 1988 National Maternal and Infant Health Survey (NMIHS). This national stratified, systematic "follow-back" survey augments information from birth records in 1988 by obtaining information on social, demographic, and economic variables from women that delivered a baby in 1988. We estimate a proportional discrete time hazard model of transitions to divorce/separation. RESULTS: Parents of a VLBW infant have 2-fold higher odds of divorce/separation compared with parents of a child with a birth weight greater than 1500 grams. Two years after delivery of a non-VLBW baby 95 percent of the marriages remain stable, while about 90 percent of the marriages remain stable following the birth of a VLBW baby. If the pregnancy was not desired, then only 85 percent of the marriages remain stable 2 years following the delivery of a VLBW infant. CONCLUSIONS: There is an evident need to counsel and support families with VLBW infants on mechanisms to cope with the initial stressors that can be anticipated to arise. PMID- 17109224 TI - Gender nonconformity and psychological distress in lesbians and gay men. AB - Some lesbians and gay men tend to be more gender nonconforming, on average and for certain traits, than their heterosexual counterparts. Gender nonconformity in childhood has also been linked to adult homosexuality. Studies of both lesbians and gay men also find elevated rates of psychological distress. We hypothesized that these facts may be related. Individuals who violate social norms for gender appropriate behavior may suffer from stigmatization by both heterosexual and homosexual people, leading to higher levels of psychological distress. We examined whether several measures of gender nonconformity were related to psychological distress in a community-based sample of gay men and lesbians. These included self-reports of childhood and adulthood gender nonconformity, as well as observer ratings of current behavior. Several measures of gender nonconformity were related to each other for both lesbians and gay men. In addition, gender nonconformity was related to psychological distress, but only for gay men. Finally, both lesbian and gay male participants reported more positive attitudes towards gender conformity than nonconformity, although the pattern was somewhat different for each group. We discuss the implications of these results for future studies of gender nonconformity and for the promotion of psychological health in lesbians and gay men. PMID- 17109225 TI - Patient-reported complications and functional outcomes of male-to-female sex reassignment surgery. AB - This study examined preoperative preparations, complications, and physical and functional outcomes of male-to-female sex reassignment surgery (SRS), based on reports by 232 patients, all of whom underwent penile-inversion vaginoplasty and sensate clitoroplasty, performed by one surgeon using a consistent technique. Nearly all patients discontinued hormone therapy before SRS and most reported that doing so created no difficulties. Preoperative electrolysis to remove genital hair, undergone by most patients, was not associated with less serious vaginal hair problems. No patients reported rectal-vaginal fistula or deep-vein thrombosis and reports of other significant surgical complications were uncommon. One third of patients, however, reported urinary stream problems. No single complication was significantly associated with regretting SRS. Satisfaction with most physical and functional outcomes of SRS was high; participants were least satisfied with vaginal lubrication, vaginal touch sensation, and vaginal erotic sensation. Frequency of achieving orgasm after SRS was not significantly associated with most general measures of satisfaction. Later years of surgery, reflecting greater surgeon experience, were not associated with lower prevalence rates for most complications or with better ratings for most physical and functional outcomes of SRS. PMID- 17109226 TI - Possible gender identity disorder in an extremely religious Muslim family. PMID- 17109228 TI - Commentary on Puts' (2006) review of The Case of the Female Orgasm: Bias in the Science of Evolution. PMID- 17109229 TI - No such thing as excessive levels of sexual behavior. PMID- 17109232 TI - The playmate and play style preferences structured interview: a comparison of children with gender identity disorder and controls. AB - The present study compared the sex-typed preferences for playmates and play styles in children referred for concerns about their gender identity development (199 boys, 43 girls) with that of controls (96 boys, 38 girls). Each child was administered the Playmate and Play Style Preferences Structured Interview (PPPSI) developed by Alexander and Hines (Alexander, G. M., & Hines, M. (1994). Child Development, 65, 869-879). In the two single dimension conditions (playmates and play styles), the controls significantly preferred same-sex playmates and same sex play styles whereas the gender-referred children significantly preferred cross-sex playmates and cross-sex play styles. Effect sizes ranged from 1.56 2.78. In the conflict condition (which required a choice between same-sex playmates and cross-sex play styles vs. cross-sex playmates and same-sex play styles), there was a general indication of a hierarchical preference for the preferred play style in the single dimension condition as opposed to the preferred playmate except for the gender-referred boys, who showed an inverted pattern. For the gender-referred group, the PPPSI data were significantly correlated with other measures of sex-typed behavior, providing evidence of predictive validity. The PPPSI also discriminated between probands threshold and subthreshold for the diagnosis of gender identity disorder. The results were discussed in relation to both basic and applied issues in the assessment of sex typed behavior in children. PMID- 17109233 TI - Minority stress and sexual problems among African-American gay and bisexual men. AB - Minority stress, such as racism and gay bashing, may be associated with sexual problems, but this notion has not been examined in the literature. African American gay/bisexual men face a unique challenge in managing a double minority status, putting them at high risk for stress and sexual problems. This investigation examined ten predictors of sexual problems among 174 African American gay/bisexual men. Covarying for age, a forward multiple regression analysis showed that the measures of self-esteem, male gender role stress, HIV prevention self-efficacy, and lifetime experiences with racial discrimination significantly added to the prediction of sexual problems. Gay bashing, psychiatric symptoms, low life satisfaction, and low social support were significantly correlated with sexual problems, but did not add to the prediction of sexual problems in the regression analysis. Mediation analyses showed that stress predicted psychiatric symptoms, which then predicted sexual problems. Sexual problems were not significantly related to HIV status, racial/ethnic identity, or gay identity. The findings from this study showed a relationship between experiences with racial and sexual discrimination and sexual problems while also providing support for mediation to illustrate how stress might cause sexual problems. Addressing minority stress in therapy may help minimize and treat sexual difficulties among minority gay/bisexual men. PMID- 17109234 TI - Single-session motivational enhancement counseling to support change toward reduction of HIV transmission by HIV positive persons. AB - Given the enormous successes in treating HIV disease with antiretroviral therapies, there is a burgeoning population of healthy, sexually active HIV+ men and women. Because HIV prevention counseling has focused traditionally on persons at risk of becoming infected, there is an urgent mandate to explore ways to engage HIV+ persons in transmission risk reduction counseling. Using two case examples, this article presents an overview of motivational interviewing in a single counseling session as a promising treatment for addressing ambivalence about safer sex with HIV+ persons. PMID- 17109235 TI - Genetic and environmental influences on cross-gender behavior and relation to behavior problems: a study of Dutch twins at ages 7 and 10 years. AB - The aim of this study was to investigate the prevalence of cross-gender behavior during childhood, to estimate the influence of genotype and environment on variation in cross-gender behavior, and to explore the association of cross gender behavior with maternal ratings of behavior problems as indexed by the Internalizing and Externalizing scales of the Child Behavior Checklist (CBCL). Cross-gender behavior was assessed by two items from the CBCL: "behaves like opposite sex" and "wishes to be of opposite sex." As part of an ongoing longitudinal study of the Netherlands Twin Registry, mothers were asked to complete the CBCL for their twins when they were 7 (n approximately 14,000 twins) and 10 years old (n approximately 8,500 twins). The prevalence of cross-gender behavior (as measured by maternal report of behaving like or wishing to be the opposite sex) was 3.2% and 5.2% for 7-year-old boys and girls, respectively, and decreased to 2.4% and 3.3% for 10-year-old boys and girls. Surprisingly, the prevalence rate of cross-gender behavior of girls with a male co-twin was lower than of girls with a female co-twin. At both ages, the similarity for cross gender behavior was greater in monozygotic than in dizygotic twins pairs. Genetic structural equation modeling showed that 70% of the variance in the liability of cross-gender behavior could be explained by genetic factors, at both ages and for both sexes. Cross-gender behavior was associated with higher scores on Internalizing and Externalizing problems, both in boys and in girls. PMID- 17109236 TI - In the mood for love or vice versa? Exploring the relations among sexual activity, physical affection, affect, and stress in the daily lives of mid-aged women. AB - How do physical affection, sexual activity, mood, and stress influence one another in the daily lives of mid-aged women? Fifty-eight women (M age, 47.6 yrs) recorded physical affection, several different sexual behaviors, stressful events, and mood ratings every morning for 36 weeks. Using multilevel modeling, we determined that physical affection or sexual behavior with a partner on one day significantly predicted lower negative mood and stress and higher positive mood on the following day. The relation did not hold for orgasm without a partner. Additionally, positive mood on one day predicted more physical affection and sexual activity with a partner, but fewer solo orgasms the following day. Negative mood was mostly unrelated to next-day sexual activity or physical affection. Sexual orientation, living with a partner, and duration of relationship moderated some of these effects. Results support a bidirectional causal model in which dyadic sexual interaction and physical affection improve mood and reduce stress, with improved mood and reduced stress in turn increasing the likelihood of future sex and physical affection. PMID- 17109237 TI - A population-based needs assessment for mental health services. AB - A needs assessment was done to objectively evaluate client need and determine gaps between these needs and the services provided by mental health services. The services included were standard case management, assertive case management and residential care across the Fraser Health Region in British Columbia, Canada. Assessments involved collection of data on current symptoms, functioning, and service use and needs. Results showed many specific areas where client needs were under met. A level-of-care planning model was applied to the data and results suggest that a high percentage of clients may be in fact receiving too high or too low a level of care for their needs. This method of performing a population based needs assessment provided findings that have proved useful for planning purposes in order to ensure that community mental health services better match client need. PMID- 17109239 TI - Obstructive sleep apnea syndrome in Prader-Willi Syndrome: an unrecognized and untreated cause of cognitive and behavioral deficits? AB - Prader-Willi Syndrome (PWS) is a rare genetic disorder characterized by a range of physical, psychological, and physiological abnormalities. It is also distinguished by the high prevalence of obstructive sleep apnea syndrome (OSAS), i.e., repetitive upper airway collapse during sleep resulting in hypoxia and sleep fragmentation. In non-PWS populations, OSAS is associated with a range of neurocognitive and psychosocial deficits. Importantly, these deficits are at least partly reversible following treatment. Given the findings in non-PWS populations, it is possible that OSAS may contribute to neurocognitive and psychosocial deficits in PWS. The present review examines this possibility. While acknowledging a primary contribution from the primary genetic abnormality to central neural dysfunction in PWS, we conclude that OSAS may be an important secondary contributing factor to reduced neurocognitive and psychosocial performance. Treatment of OSAS may have potential benefits in improving neurocognitive performance and behavior in PWS, but this awaits confirmatory investigation. PMID- 17109240 TI - Nonpulsatile and noninvasive transmittance and reflectance tissue-bed oximetry. AB - A new optical device was developed that measures blood pressure noninvasively, in small human subjects (neonates and premature infants) and small animals (Roeder RAR. Transducer for indirect measurement of blood pressure in small human subjects and animals, Purdue University, BME; 2003.: xi, 50 p.). The ability of this device to measure oxygen saturation enhances its value. The objective of this research was to add the ability to obtain SaO(2) from the same device and to obtain the calibration curve. Another objective was to determine which measurement method (transmittance or reflectance) is preferable. This new oximeter is unlike the conventional pulse oximeter in that it does not require a pulse, making it ideal for measuring oxygen saturation noninvasively in small human subjects with small amplitude pulses or without a pulse. A study was performed in 11 pigs, ranging in weight 20-27 kg. The pig tail was used as the measuring site for %SaO(2) measurements. Arterial blood samples were obtained from the femoral artery and oxygen saturation was measured with a blood-gas analyzer. A small blood-pressure cuff was used to render the optical path bloodless. A comparison of the transmittance and reflectance methods for measuring oxygen saturation was made. %SaO(2) measurements ranged from 4% to 100%. It was found that both the transmittance and reflectance methods can be used to measure %SaO(2) reliably in situations with or without a pulse. PMID- 17109238 TI - In your right mind: right hemisphere contributions to language processing and production. AB - The verbal/nonverbal account of left and right hemisphere functionality is the prevailing dichotomy describing the cerebral lateralization of function. Yet the fact that the left hemisphere is the superior language processor does not necessarily imply that the right hemisphere is completely lacking linguistic ability. This paper reviews the growing body of research demonstrating that, far from being nonverbal, the right hemisphere has significant language processing strength. From prosodic and paralinguistic aspects of speech production, reception, and interpretation, to prelexical, lexical and postlexical components of visual word recognition; strong involvement of the right hemisphere is implicated. The evidence reviewed challenges the notion that language is solely a function of the "verbal" left hemisphere, indicating that the right cerebral hemisphere makes significant and meaningful contributions to normal language processing as well. PMID- 17109241 TI - The role of morphology and short vowelization in reading Arabic among normal and dyslexic readers in grades 3, 6, 9, and 12. AB - This study was an investigation of several Arabic reading measures among dyslexics and normal Arabic readers across different ages (grades 3, 6, 9, and 12): the role of morphology, short vowelization (phonological and syntactic skills), spelling, reading isolated words, and reading comprehension. The results of the one-way ANOVAs indicated clear differences between the dyslexic readers and the normal readers on all reading measures. However, the stepwise regression analysis revealed consistent orthographic results: morphology (identification and/or production) and spelling were generally the most powerful predictors of both reading accuracy and reading comprehension among dyslexic and normal readers across these different age groups. The results are discussed in terms of the characteristics of the Arabic orthography and the heavy reliance of readers at all levels and ages on orthographic factors in reading. PMID- 17109242 TI - The effect of language immersion education on the preattentive perception of native and non-native vowel contrasts. AB - Proficiency in a second language (L2) may depend upon the age of exposure and the continued use of the mother tongue (L1) during L2 acquisition. The effect of early L2 exposure on the preattentive perception of native and non-native vowel contrasts was studied by measuring the mismatch negativity (MMN) response from 14 year-old children. The test group consisted of six Finnish children who had participated in English immersion education. The control group consisted of eight monolingual Finns. The subjects were presented with Finnish and English synthetic vowel contrasts. The aim was to see whether early exposure had resulted in the development of a new language-specific memory trace for the contrast phonemically irrelevant in L1. The results indicated that only the contrast with the largest acoustic distance elicited an MMN response in the Bilingual group, while the Monolingual group showed a response also to the native contrast. This may suggest that native-like memory traces for prototypical vowels were not formed in early language immersion. PMID- 17109243 TI - [Prehypertension--a definition]. PMID- 17109244 TI - [Proven and new parameters of hypertensive target organ damage]. PMID- 17109245 TI - [Normokalemic primary hyperaldosteronism]. PMID- 17109246 TI - [Adherence to medical treatment]. PMID- 17109247 TI - [The Stamp of Quality (Prufsiegel) of the German Hypertension League for the clinical validation of blood pressure measuring devices. Results from the testing of 51 devices]. PMID- 17109249 TI - [Antithrombotic treatment of acute coronary syndromes without ST-segment elevation]. PMID- 17109250 TI - [Differentiated treatment of diabetics with an acute coronary syndrome]. PMID- 17109251 TI - [Risk stratification in the acute coronary syndrome. Diabetes--a diagnostic dilemma]. PMID- 17109252 TI - [Pathogenesis and therapy of the acute coronary syndrome: differentiated advantage of glycoprotein IIb/IIIa receptor antagonists for high-risk patients]. AB - Pathogenesis and therapy of the acute coronary syndrome: differentiated advantage of glycoprotein IIb/IIIa receptor antagonists for high-risk patients Activated thrombocytes and their aggregation into a thrombus play a decisive role in the pathogenesis of the acute coronary syndrome. Glycoprotein (GP) IIb/IIIa receptor antagonists block the binding of fibrinogen to GP IIb/IIIa receptors of activated thrombocytes and inhibit their aggregation. Patients with an acute coronary syndrome and raised troponin levels who are to undergo primary coronary revascularization profit particularly from treatment with GPIIb/IIIa receptor antagonists. This is especially so if they also have diabetes. PMID- 17109253 TI - Investigation of cytotoxic activity on human cancer cell lines of arborinine and furanoacridones isolated from Ruta graveolens. AB - The cytotoxic effects of a series of furanoacridones isolated from Ruta graveolens L. (Rutaceae) and of two further acridone alkaloids (arborinine and evoxanthine) were investigated by means of the MTT assay, using the human cell lines HeLa, MCF7 and A431. Arborinine proved best in inhibiting the proliferation of all three cell lines. The cytotoxic potency of the furacridone alkaloids was a function of their lipid solubility, which was determined by means of PAMPA. The capacity of the most effective furanoacridones to induce apoptosis was demonstrated by flow cytometric cell cycle analysis and by staining with ethidium bromide and acridine orange. This finding was reinforced by determining the apoptosis-regulating factors Bcl-2 and Bax, which were revealed by means of RT PCR to change dose-dependently. The data presented here indicate that naturally occurring furanoacridones can be regarded as excellent starting structures for the potential development of new anticancer agents. PMID- 17109254 TI - Protein tyrosine nitration induced by heme/hydrogen peroxide: inhibitory effect of hydroxycinnamoyl conjugates. AB - The present study was designed to optimize the experimental conditions that govern the heme-catalyzed nitration of protein tyrosine residues by nitrite, and, within this framework, to study the effects of 3,5-dicaffeoylquinic acid and its methyl ester, both of which have been previously reported to be antioxidants and inhibitors of leukocyte functions. Although the presence of hydrogen peroxide is essential in cell-free systems, an excess of this compound was found to be detrimental, so much so that an increase in hemin concentration actually resulted in an inverse effect on the reaction, depending on the levels of fixed hydrogen peroxide. Unlike previous reports on nitrite-induced albumin tyrosine nitration, the optimal pH here was found to be 7.0. The two caffeoyl conjugates tested were found to be effective inhibitors of protein nitration, with IC50 values ranging from 20 - 30 microM, regardless of the presence of bicarbonate. For the inhibition of myeloperoxidase-catalyzed protein nitration by human polymorphonuclear leukocytes stimulated by phorbol ester, the potencies obtained were up to two times higher. This is the first time that caffeoylquinic esters have been reported as inhibitors of heme-based protein nitration. PMID- 17109255 TI - Molecular authentication of Alisma orientale by PCR-RFLP and ARMS. AB - As a widely used medicinal plant, Alisma orientale is always a possible target for fraudulent labeling. The internal transcribed spacer (ITS) regions of nuclear ribosomal DNA (nrDNA) of the six species of genus Alisma were sequenced, and two variant sites were found to be specific for A. orientale. Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis and amplification refractory mutation system (ARMS) analysis were applied to the ITS region for the identification of A. orientale. A restriction site for PSTI useful for PCR-RFLP analysis was detected and a pair of diagnostic primers DFZX-JB02S and DFZX-JB02X were designed for ARMS. PMID- 17109256 TI - Vitamin E reduces antidepressant-related beta-adrenoceptor down-regulation in cultured cells. Comparable effects on St. John's wort and tricyclic antidepressant treatment. AB - The mode of action of antidepressants is still a matter of debate. Acute inhibition of neurotransmitter reuptake in central neuronal synapses, followed by a down-regulation of central postsynaptic beta-adrenoceptor (beta-AR) numbers were consistently observed in vivo, while a reduction in surface beta-AR density was found in cell cultures. Effects of the tricyclic antidepressant desipramine (DMI) were abolished by vitamin E (alpha-TOC) in vitro as well as in vivo. Alpha TOC interfered with antidepressant-induced changes of cellular plasma membrane properties and with recycling of beta-AR. St. John's wort (SJW) extract reduced beta-AR numbers in cultured cells to a similar extent as DMI or the selective serotonin re-uptake inhibitor fluoxetine. We chronically co-exposed cell cultures to SJW extract and to alpha-TOC. Receptor down-regulation following exposure to the plant extract was inhibited in the presence of alpha-TOC suggesting a mode of action of SJW extract comparable to that of synthetic antidepressants. Inhibition of cell proliferation by the plant extract was also significantly reduced by alpha-TOC. PMID- 17109263 TI - [Frequency of cytological procedures in diagnostic bronchoscopy of peripheral pulmonary modules and masses]. AB - BACKGROUND: The combination of forceps biopsy and cytological methods improves the yield in diagnostic bronchoscopy of peripheral lung cancer. Transbronchial catheter aspiration seems to be an underutilized cytological technique, which is valuable and minimally invasive. We inquired for the frequency of cytological procedures and the importance of transbronchial catheter aspiration in comparison to transbronchial needle aspiration and bronchial brushing. METHODS: We sent questionnaires to 99 clinically practising and educating pulmonologists. RESULTS: The response amounted to 69 %. Nearly 79 % of the pulmonologists usually combine forceps biopsy with cytological procedures. Instead of 51 % bronchial brushing, transbonchial catheter aspiration as well as needle aspiration is the most applied cytological technique by only 28 % of the interrogated physicians. 47 % of the investigators practise bronchial washing and 27 % practise core roll preparations as a rule. CONCLUSIONS: Most of the specialized clinics in respiratory medicine in Germany add cytological methods to transbronchial forceps biopsy. This practice is pregnant with meaning because of a higher diagnostic yield while accuracy is likewise high. Unlike bronchial brushing transbronchial catheter aspiration and needle aspiration are underutilized for different reasons in spite of the fact, that these are the only procedures with a significant higher sensitivity in comparison to forceps biopsy. PMID- 17109264 TI - [Gastrointestinal tuberculosis as the main manifestation of systemic tuberculosis]. AB - We describe a rare case of tuberculosis with mainly gastrointestinal problems. The 52-year-aged female patient came to hospital with unclear pain in the lower abdomen and ascites that was refractory to therapy. The computed tomography of the thorax showed right-sided confluating lymphoid nodes, the CT of the abdomen showed ascites and nodular structures near the coecum. Tissue samples were taken from the mucosa of the colon, the inflammatory altered peritoneum, the left bronchus of the upper lobe and the confluating lymphoid nodes in the mediastinum during colonoscopy, diagnostic laparoscopy and bronchoscopy. The samples from the peritoneum showed granulomas with caseating necroses in histological slices. Mycobacterium tuberculosis was detected by PCR in the tissue samples from the lymphoid tissue of the mediastinum. Furthermore, Mycobacterium tuberculosis grew in cultures from samples of the abdominal ascites. The symptoms and pathological findings improved under a therapy comprising isoniazid, rifampicin, ethambutol and pyrazinamid. PMID- 17109265 TI - [Clinical significance of gastrooesophageal reflux in asthma]. AB - In clinical practice, gastrooesophageal reflux (GER) and bronchial asthma often coincide. In the case of sufficient asthma control, the asthma does not need to be taken into account when treatment of GER is being evaluated. In patients with symptomatic asthma despite adequate antiasthmatic treatment, a possible causal relationship between GER and the poor responsiveness to asthma therapy has to be considered. An algorithm to guide the diagnostic and therapeutic steps in such cases is presented. PMID- 17109266 TI - [Chronic obstructive pulmonary disease: basic features, risk factors and diagnosis]. PMID- 17109267 TI - [Pharmacotherapy of severe acute respiratory syndrome (SARS)]. AB - Severe acute respiratory syndrome (SARS) constitutes the first new infectious disease of the current millennium. It is caused by the novel SARS-Coronavirus (SARS-CoV). SARS is related to a high morbidity and mortality and first appeared during an epidemic in 2002 - 2003. To date no specific therapy against the SARS CoV is available. Due to the rapid spread of SARS during the epidemics in 2002 - 2003, randomised and controlled multicentre studies were not performed. Therefore, general guidelines have not been developed. Since the outbreak, scientists have been testing potential antiviral substances using in vitro and animal models. This study analyses the presently available in vitro and in vivo data on the pharmacotherapy of SARS. PMID- 17109268 TI - [Christmas seals]. AB - Christmas seals, i. e., special stamps used to decorate or seal Christmas and New Year's mail, were created by the Danish post office clerk E. Holboell. The proceeds from the sale of the stickers were meant to alleviate the suffering of children sick with tuberculosis. The first Christmas seal, showing a portrait of Queen Louise of Denmark, was issued on December 10, 1904. The demand at the post office was enormous. The funds raised exceeded all expectations and made it possible to finance the construction of a sanatorium for tuberculosis children. The idea of Christmas seals spread quickly around the world and ended up being copied in 130 countries. At the beginning of the 20 (th) century, the small stamp with the red double-barred cross became a banner for the crusade against tuberculosis. For many patients, it also represented a symbol of hope for recovery even though this hope, given the cure rates of a sanatorium treatment, only became a reality for a few. Worldwide, Christmas seals were and are colourful and imaginatively designed, mostly with Christmas symbols or motives relating to the fight against tuberculosis. The funds raised through the sale of the seals initially helped to build hospitals and were later also used to screen persons at risk for tuberculosis, to improve housing conditions of patients and for other measures of support. With the decrease of tuberculosis, some organisations discontinued fundraising through Christmas seals while others widened their intended purpose and supported, for example, the prevention of and research on lung diseases. In Germany, Christmas seals are closely connected with the German Central Committee against Tuberculosis. Today the "Kuratorium Tuberkulose in der Welt" is the only organisation in this country that still raises funds by this means. The funds are mostly used to assist developing countries with a high burden of tuberculosis. PMID- 17109269 TI - [Ca2+-signaling in smooth muscles cells of the airways in T-bet knock-out mice]. AB - BACKGROUND: Airway smooth muscle cells (ASMC) play a key role in bronchial hyperresponsiveness (BHR). A major component of the signalling cascade leading to ASMC contraction is calcium. T-bet knock-out (KO) mice show the key features of allergic asthma such as a shift towards T (H2)-lymphocytes and display a broad spectrum of asthma-like histological and functional characteristics. In this study, we aimed at investigating whether Ca (2+)-homeostasis of ASMC is altered in T-bet KO-mice as an experimental model of asthma. METHODS: Lung slices of 100 to 200 microm thickness were obtained from T-bet KO- and wild-type mice. Airway contractions in response to acetylcholine (ACH) were measured by video-microscopy and Ca (2+)-signaling in single ASMC of lung slices was assessed using two-photon microscopy. RESULTS: Airways from T-bet KO-mice showed increased baseline airway tone (BAT) and BHR compared to those of wild-type mice. The increased BAT was correlated with an increased incidence of spontaneous changes in intracellular Ca (2+)-concentrations, whereas BHR correlated with higher ACH-induced Ca (2+) transients and an increased proportion of ASMC showing Ca (2+)-oscillations. Emptying intracellular Ca (2+)-stores using caffeine or cyclopiazonic acid induced higher Ca (2+)-elevations in ASMC from T-bet KO compared to wild-type mice. CONCLUSIONS: Altered Ca (2+)-homeostasis of ASMC contributes to increased BAT and BHR in lung slices from T-bet KO mice as a murine asthma model. We propose that a higher Ca (2+)-content of the intracellular Ca (2+)-stores is involved in the pathophysiology of these changes. PMID- 17109270 TI - [Recommendations for planning and realisation of exercise training in outpatient lung sports groups]. AB - Asthma and COPD are common diseases inducing high costs. In national and international guidelines exercise training is stated as the main component amongst the nonmedical treatment options. With exercise training beneficial effects in exercise performance, quality of life and health-related costs can be obtained. Options for exercise training all over the country should be established. Therefore not only inpatient, but also outpatient structures, especially lung sports groups have to be organized to enable a continuous training. For this special way of long term training no systematic guidelines do exist. Further aggravating factors are the different basic vocational education of the trainers and the heterogeneity of the participants. The intention of the authors was to develop recommendations for a structured training and training sensitive to the target group. The realisation of training with this approach should approximate training to evidence based knowledge and contribute to the quality of training patients with lung diseases. PMID- 17109272 TI - [Prognosis after complete surgical resection for non-small cell lung cancer based on the staging classification]. AB - BACKGROUND AND OBJECTIVE: The importance of accurate staging according to the international TNM staging system of non-small cell lung cancer (NSCLC) for patient management and ascertaining individual prognosis cannot be overemphasized. The TNM classification is scheduled to be revised in 2007. In a large single-center collective we investigated the prognosis for patients who had complete resection of a NSCLC. PATIENTS AND METHODS: We retrospectively reviewed hospital records and follow-up data of 2,378 patients operated on between 1996 and 2005 for NSCLC. Complete resection was achieved in 2,083 patients. Systematic hilar and mediastinal lymph node dissection was performed concurrently. Probability of survival was then analysed with the Kaplan-Meier method. The significance of differences between subgroups was calculated using the log-rank test. Odds ratios with 95 % confidence intervals (CI) were calculated for each characteristic. The Cox model was used for multivariate analyses. RESULTS: The 5 year survival for patients after complete resection was 50.7 %. The 5-year survival rates for clinical stages were 72 % for stage IA, 59.8 % for stage IB, not defined for stage IIA, 47,8 % for stage IIB, 45 % for stage IIIA, 38.7 % for stage IIIB, and not defined for stage IV. There were significant differences in survival between stages IIIB and IV (p = 0.013). There was a trend towards significance between patients with IA and IB (p = 0.052). However, there was no significant difference between patients with all the other stages. 5-year survival according to pathological stages was: stage IA 68.5 %; stage IB 66.6 %; stage IIA 55.3 %; stage IIB 49.0 %; stage IIIA 35.8 %; stage IIIB 35.4 %; stage IV not defined. Gender, age and type of histology were found by multivariate analysis to be significant independent prognostic factors for survival. CONCLUSIONS: The TNM and stage grouping classification is valid for defining prognosis and prognosis-related criteria in patients with NSCLC. The difference in prognosis between clinical stages IIIB and IV was significant, but not that between all the other related subgroups. Concordance with histological staging demonstrated the quality of existing clinical staging methods and related strategies. Complete surgical resection, age, gender, histology and stage of the disease significantly influenced long-term survival. PMID- 17109273 TI - [Postoperative infection of an epigastric hematoma caused by Aeromonas veronii biovar sobria]. AB - HISTORY: A 52-year-old man with a gastric signet-ring cell carcinoma but without acute symptoms was admitted for reconstructive surgery of the gastrointestinal tract. INVESTIGATIONS: Before the present surgery all functional and radiological tests merely confirmed the previously known disease. Except for mild anemia and abnormal electrolytes all laboratory tests were within normal limits. COURSE, DIAGNOSIS AND TREATMENT: The patient underwent reconstructive surgery of the gastrointestinal tract, namely an ascending sigmoidostomy and resection of an enterocutaneous fistula. For a few days the postoperative development was as expected and the drain, placed during surgery, was removed at the expected time. 9 days postoperatively the patient developed signs of an infection (fever up to 38.8 degrees C, increased WBC and raised C-reactive protein levels). Computed tomography (CT) of the abdomen revealed an epigastric tumor measuring 6 x 5 cm. CT-guided needle aspiration of this lesion showed macroscopic signs of an infected hematoma. A pigtail catheter was successfully implanted for continuous drainage. Both the fluids obtained from CT-guided aspiration and the pigtail drain grew Aeromonas veronii biovar sobria when cultured on standard blood agar. Administration of both cefotaxim and metronidazole for 10 days produced a decrease in the inflammatory parameters. The abdominal CT at that time showed a noticeable regression of the epigastric mass so that the patient was discharged from hospital 3 weeks after surgery. CONCLUSION: This case emphasizes the importance of adequately dosed antibiotic therapy, also for unusual bacteria such as species of Aeromonas. PMID- 17109274 TI - [Primary adrenal cortical insufficiency (Addison's disease)]. PMID- 17109275 TI - [Immunobiology of chronic inflammatory bowel diseases]. PMID- 17109276 TI - [Nosocomial pneumonia: diagnosis]. PMID- 17109277 TI - [Subclinical albuminuria, microalbuminuria and proteinuria--accepted cardiovascular risk markers?]. PMID- 17109278 TI - [Diastolic heart failure--what a "non-cardiologist" should know]. PMID- 17109279 TI - [Recommendations for platelet transfusion by the Joint Thrombocyte Working Party of the German Societies of Transfusion Medicine and Immunohaematology (DGTI), Thrombosis and Haemostasis Research (GTH), and Haematology and Oncology (DGHO)]. PMID- 17109281 TI - [Availability of tobacco and alcohol become limited]. PMID- 17109282 TI - Prospective studies in patients with intraventricular haemorrhage without the capacity to give consent in Germany--a legal dilemma. AB - OBJECTIVE: The prognosis of intracranial haemorrhage with ventricular participation is poor. The acute onset of hydrocephalus has to be treated with an external ventricular drainage. However, intraventricular blood clots often obstruct the drainage catheter; the reinsertion is usually associated with an increased risk of complications. Therefore, intraventricular thrombolysis using recombinant tissue plasminogen activator (rt-PA), urokinase or streptokinase has been performed in several cases. In Germany, rt-PA does not have approval for intraventricular applications, and the risks and benefits of this treatment are as yet unclear. Several authors recommend its use only in the frame of prospective studies, meaning that intraventricular administration of thrombolytic agents within an individual therapy trial could be viewed as medical malpractice. METHODS: We designed a national prospective randomised and controlled study in patients with intraventricular haemorrhage to investigate the risks and effects of intraventricular rt-PA treatment. The local ethics committee and lawyers did not accept the study protocol due to a non-acceptable risk of complications. In two further conferences, the risk of undertreatment in the control group was a major reason not to accept the protocol. We discuss the conflicts of law in related studies. CONCLUSIONS: There is a need for a solution to the legal conflicts of medical malpractice, unclear risk-benefit ratio and unfeasibility due to risk of complications or undertreatment in studies with patients without the capacity to give informed consent. PMID- 17109283 TI - Interleukin-2: a potential treatment option for postherpetic neuralgia? AB - Postherpetic neuralgia is a chronic pain syndrome that is often difficult to treat and can lead to a disabling disease if it is resistant to therapy. Presented here is the case of a 46-year-old patient with human immunodeficiency virus infection and chronic, treatment-resistant neuralgia. Postherpetic pain resolved after treatment with 1 cycle of subcutaneous recombinant interleukin-2. PMID- 17109284 TI - The practice of travel medicine: guidelines by the Infectious Diseases Society of America. PMID- 17109285 TI - Clinical reevaluation of the QuantiFERON TB-2G test as a diagnostic method for differentiating active tuberculosis from nontuberculous mycobacteriosis. AB - INTRODUCTION: We reevaluated the usefulness of a whole-blood interferon-gamma enzyme-linked immunosorbent assay (QuantiFERON TB-2G [QFT-TB]; Cellestis) in obtaining a differential diagnosis between active tuberculosis (TB) and nontuberculous mycobacteriosis (NTM). METHODS: The subjects were 50 healthy volunteers, 50 patients with active TB, and 100 patients with NTM who satisfied the diagnostic guidelines of the American Thoracic Society from April 2005 through June 2006. The tuberculin skin test (TST) and the QFT-TB test were performed for all subjects. The QFT-TB test was performed every 2 months. RESULTS: Of the healthy volunteers, 64% had a negative TST result and 94% had a negative QFT-TB test result. Of the patients with active TB, 64% had a positive TST result and 4% had a negative QFT-TB test result. Of the patients with pulmonary Mycobacterium avium complex disease, 60% had a positive TST result and 7% had a positive QFT-TB test result. The QFT-TB test had a mean sensitivity of 86% and a mean specificity of 94%. The QFT-TB test results for patients with active TB transiently decreased during treatment involving antituberculous drugs. The rate of positive QFT-TB test results was 86% at the initiation of treatment, 48% 6 months later, and 33% 12 months later. CONCLUSIONS: We confirmed that the QFT-TB test is a useful diagnostic method for differentiating active pulmonary TB from NTM, compared with the TST. However, because it is possible that the effect of the QFT-TB test may be long lasting after treatment and may not be resolved over time, even with treatment, as in this study, it may not provide any level of certainty regarding cure of infection. PMID- 17109286 TI - Tuberculin skin testing is useful in the screening for nontuberculous mycobacterial cervicofacial lymphadenitis in children. AB - BACKGROUND: We evaluated the diagnostic usefulness of tuberculin skin testing in the screening for nontuberculous mycobacterial (NTM) infection in children. METHODS: We enrolled 180 children who had chronic cervicofacial lymphadenitis in our study. Skin testing was done using antigens of Mycobacterium tuberculosis, Mycobacterium avium, Mycobacterium kansasii, and Mycobacterium scrophulaceum. The reference standard for NTM infection was a positive culture result, identification by PCR, or both. Receiver operating characteristic analysis was used to identify the optimal cutoff point in skin induration for the detection of NTM infection. Accuracy of the mycobacterial skin tests was quantified using sensitivity and specificity rates and positive and negative predictive values at the optimal skin induration cutoff. RESULTS: A total of 112 NTM infections were identified, of which 83 were caused by M. avium, 21 by Mycobacterium haemophilum, and 8 by other NTM species. At the optimal cutoff for a positive test (5 mm), tuberculin skin testing had a sensitivity and specificity of 70% and 98%, respectively, and a positive predictive value and a negative predictive value of 98% and 64%, respectively, compared with a sensitivity and a specificity of 93% and 97%, respectively; M. avium sensitin, the best-performing skin test, had positive and negative predictive values of 98% and 90%, respectively. CONCLUSION: Tuberculin skin testing could be valuable as a first step in the diagnostic analysis of cervicofacial lymphadenitis in children without a history of TB exposure or bacille Calmette-Guerin vaccination. PMID- 17109287 TI - The tuberculin skin test: a useful screen for nontuberculous mycobacterial lymphadenitis in regions with a low prevalence of tuberculosis? PMID- 17109288 TI - Characterization of multidrug-resistant influenza A/H3N2 viruses shed during 1 year by an immunocompromised child. AB - BACKGROUND: Development of influenza drug resistance is an important problem in immunocompromised children that could result in treatment failure and viral transmission to others. METHODS: A total of 17 influenza A/H3N2 isolates were recovered over a period of 1 year from an immunocompromised child who was initially treated with oseltamivir and then with amantadine and zanamivir for viral pneumonitis. Drug susceptibility phenotypes to oseltamivir, zanamivir, and peramivir were evaluated by neuraminidase (NA) inhibition assays, and sequence analysis of key viral genes (i.e., M2, NA, and hemagglutinin [HA]) was performed. The impact of NA mutations identified in oseltamivir-resistant isolates was analyzed using recombinant NA proteins. RESULTS: An influenza A variant with NA mutations E59G, E119V, and I222V was first detected after 38 days of oseltamivir treatment. In an NA inhibition assay, this variant was 274 times more resistant to oseltamivir than the original isolate but was susceptible to zanamivir. The I222V substitution enhanced the level of oseltamivir resistance that was primarily conferred by the E119V mutation in recombinant NA proteins. Remarkably, the E119V mutation persisted for 8 months after cessation of oseltamivir. Amantadine therapy led to rapid emergence of the M2 mutation S31N, which is known to confer amantadine resistance. The patient shed the virus intermittently while receiving nebulized zanamivir therapy despite the absence of a resistance phenotype, which could be the result of nonoptimal drug delivery and impaired host immunity. CONCLUSIONS: This study highlights the potential for emergence and persistence of multidrug-resistant influenza isolates in immunocompromised subjects even after cessation of treatment, reinforcing the need for development of new anti-influenza compounds. PMID- 17109289 TI - Influenza will not miss opportunities. PMID- 17109290 TI - Beyond viruses: clinical profiles and etiologies associated with encephalitis. AB - BACKGROUND: Encephalitis is a complex syndrome, and its etiology is often not identified. The California Encephalitis Project was initiated in 1998 to identify the causes and further describe the clinical and epidemiologic characteristics of encephalitis. METHODS: A standardized report form was used to collect demographic and clinical data. Serum, cerebrospinal fluid, and respiratory specimens were obtained prospectively and were tested for the presence of herpesviruses, arboviruses, enteroviruses, measles, respiratory viruses, Chlamydia species, and Mycoplasma pneumoniae. The association between an identified infection and encephalitis was defined using predetermined, organism-specific criteria for confirmed, probable, or possible causes. RESULTS: From 1998 through 2005, a total of 1570 patients were enrolled. Given the large number of patients, subgroups of patients with similar clinical characteristics and laboratory findings were identified. Ten clinical profiles were described. A confirmed or probable etiologic agent was identified for 16% of cases of encephalitis: 69% of these agents were viral; 20%, bacterial; 7%, prion; 3%, parasitic; and 1%, fungal. An additional 13% of cases had a possible etiology identified. Many of the agents classified as possible causes are suspected but have not yet been definitively demonstrated to cause encephalitis; these agents include M. pneumoniae (n=96), influenza virus (n=22), adenovirus (n=14), Chlamydia species (n=10), and human metapneumovirus (n=4). A noninfectious etiology was identified for 8% of cases, and no etiology was found for 63% of cases. CONCLUSIONS: Although the etiology of encephalitis remains unknown in most cases, the recognition of discrete clinical profiles among patients with encephalitis should help focus our efforts toward understanding the etiology, pathogenesis, course, and management of this complex syndrome. PMID- 17109291 TI - Assessment of the utility of viral culture of cerebrospinal fluid. AB - Nucleic acid amplification testing is the preferred method to detect enteroviruses and Herpesviridae in cerebrospinal fluid, but clinicians still request viral culture. Review of 22,394 viral cultures of cerebrospinal fluid samples found that <0.1% recovered nonenterovirus, non-Herpesviridae species, suggesting that, when nucleic acid amplification testing is performed, viral culture may have no additional benefit. PMID- 17109292 TI - Scedosporium infection in a tertiary care cancer center: a review of 25 cases from 1989-2006. AB - We reviewed the records of patients with cancer who had Scedosporium infection (due to Scedosporium apiospermum and Scedosporium prolificans in 21 and 4 patients, respectively). The incidence of Scedosporium infection increased from 0.82 cases per 100,000 patient-inpatient days (in 1993-1998) to 1.33 cases per 100,000 patient-inpatient days (in 1999-2005). Cases of S. prolificans infection occurred only after 2000. Dissemination occurred in 16 patients (64%). The 12 week mortality rates were 70% and 100% for S. apiospermum and S. prolificans infection, respectively. PMID- 17109293 TI - A 26-year-old woman with a rash on her extremities. PMID- 17109294 TI - The emerging clinical importance of non-O157 Shiga toxin-producing Escherichia coli. AB - In 1982, hemorrhagic colitis and hemolytic-uremic syndrome were linked to infection with Escherichia coli O157:H7, a serotype now classified as Shiga toxin producing E. coli (STEC). Thereafter, hemorrhagic colitis and hemolytic-uremic syndrome associated with non-O157 STEC serogroups were reported, with the frequency of non-O157 STEC illness rivaling that of O157:H7 in certain geographic regions. In the United States, non-O157 E. coli may account for up to 20%-50% of all STEC infections. A high index of suspicion, paired with options to test for non-O157 STEC infection, are necessary for early recognition and appropriate treatment of these infections. Supportive care without the use of antibiotics is currently considered to be optimal treatment for all STEC infections. This commentary provides a perspective on the non-O157 STEC as human pathogens, how and when the clinician should approach the diagnosis of these organisms, and the challenges ahead. PMID- 17109295 TI - Outbreaks of enteric disease associated with animal contact: not just a foodborne problem anymore. AB - In the past 10 years, an increasing number of outbreaks of enteric disease associated with animals in public settings, such as fairs and petting zoos, have been reported. Fifty-five of these outbreaks that occurred in the United States during 1991-2005 are reviewed in this article. Lessons learned from these outbreaks and recommendations for prevention are also discussed. Physicians should be aware of this important public health problem and play an active role in prevention of human illness associated with animals in public settings. PMID- 17109296 TI - Antimicrobial-associated QT interval prolongation: pointes of interest. AB - Until recently, cardiac toxicity manifesting in the form of arrhythmias related to QT interval prolongation was uncommonly appreciated within the antimicrobial class of drugs, but it was well described among antiarrhythmic agents. Antimicrobials that are associated with QT prolongation include the macrolides/ketolides, certain fluoroquinolones and antimalarials, pentamidine, and the azole antifungals. Although, in most cases, mild delays in ventricular repolarization caused by these drugs are clinically unnoticeable, they may serve to amplify the risk for torsades de pointes (TdP) when prescribed in the setting of other risk factors. Conditions or variables that influence proarrhythmic risk include sex, age, electrolyte derangements, structural heart disease, pharmacokinetic/pharmacodynamic interactions, and genetic predisposition. It is important that clinicians be knowledgeable about drugs with QT liability, as well as the risk factors that increase the probability of TdP. Additionally, because TdP remains a difficult-to-measure adverse event, we must rely upon multiple data sources to determine the risk versus the benefit for newly approved drugs. PMID- 17109297 TI - Serological evidence of HIV-associated infection among HIV-1-infected adults in Botswana. AB - In industrialized countries, it is recommended that adults with human immunodeficiency virus type 1 (HIV-1) infection undergo baseline screening for pathogens that might cause latent or active infections, such as syphilis, hepatitis B, hepatitis C, infection due to Toxoplasma gondii, and cytomegalovirus infection. A paucity of data exist from sub-Saharan Africa describing the prevalence of these pathogens. We report data for HIV-1-infected adults referred for initiation of highly active antiretroviral therapy in Botswana. PMID- 17109298 TI - Computed tomography characteristics of Burkholderia pseudomallei-associated liver abscess. PMID- 17109299 TI - Neurotoxicity of artemisinin derivatives. PMID- 17109300 TI - Role of intravenous immunoglobulin administration in Japanese encephalitis. PMID- 17109301 TI - Telithromycin and myasthenic crisis. PMID- 17109302 TI - Buprenorphine and HIV primary care: new opportunities for integrated treatment. AB - Drug abuse and infection with human immunodeficiency virus (HIV) are associated with high rates of morbidity and mortality, but, because of medical, social, and legal factors, opiate addiction/dependence is a major obstacle to successful treatment of disease--for example, treatment of acquired immunodeficiency syndrome (AIDS) with highly active antiretroviral therapy. In an effort to improve the opportunity for treatment of drug abuse and HIV infection, the Forum for Collaborative HIV Research, in collaboration with the Substance Abuse and Mental Health Services Administration, the National Institute on Drug Abuse, the Centers for Disease Control and Prevention, and other agencies, presented a workshop entitled "Buprenorphine in the Primary HIV Care Setting." Participants reviewed and discussed current issues, such as the introduction of and sources for the provision of buprenorphine in HIV primary care settings and strategies for integrating treatment of HIV-infected drug abusers, all of which are covered in this supplement. PMID- 17109303 TI - Opioid dependence: rationale for and efficacy of existing and new treatments. AB - Opioid dependence is a chronic and relapsing medical disorder with a well established neurobiological basis. Opioid agonist treatments, such as methadone and the recently approved buprenorphine, stabilize opioid receptors and the intracellular processes that lead to opioid withdrawal and craving. Both methadone and buprenorphine have been proven effective for the treatment of opioid dependence and can contribute to a decreased risk of human immunodeficiency virus (HIV) transmission. In addition, a buprenorphine/naloxone combination appears to have a decreased potential for abuse or diversion, compared with that associated with methadone. Largely because of these properties, recent legislation now affords an unprecedented opportunity for general physicians to offer opioid agonist treatment through their offices. This review focuses on the neurobiological basis of opioid dependence, the rationale for methadone and buprenorphine treatments, and issues in prescribing these medications to patients with HIV infection. PMID- 17109304 TI - The potential role of buprenorphine in the treatment of opioid dependence in HIV infected individuals and in HIV infection prevention. AB - Untreated opioid dependence is a major obstacle to the successful treatment and prevention of human immunodeficiency virus (HIV) infection. In this review, we examine the interwoven epidemics of HIV infection and opioid dependence and the emerging role of buprenorphine in improving HIV treatment outcomes among infected individuals, as well as its role in primary and secondary prevention. This article addresses some of the emerging issues about integrating buprenorphine treatment into HIV clinical care settings and the various strategies that must be considered. Specifically, it addresses the role of buprenorphine in improving HIV treatment outcomes through engagement in care, access to antiretroviral therapy and preventive therapies for opportunistic infections, and the potential benefits of and pitfalls in integrating buprenorphine into HIV clinical care settings. We discuss the key research questions regarding buprenorphine in the area of improving HIV treatment outcomes and prevention, including a review of published studies of buprenorphine and antiretroviral treatment and currently ongoing studies, and provide insight into and models for integrating buprenorphine into HIV clinical care settings. Dialogue among practitioners and policy makers in the HIV care and substance abuse communities will facilitate an effective expansion of buprenorphine and ensure that these beneficial outcomes are achieved. PMID- 17109305 TI - A trial of integrated buprenorphine/naloxone and HIV clinical care. AB - BACKGROUND: Untreated opioid dependence adversely affects the care of human immunodeficiency virus (HIV)-positive patients. Buprenorphine, a partial opioid agonist, is available for maintenance treatment of opioid dependence in HIV specialty settings. We investigated the feasibility and efficacy of integrating buprenorphine, along with 2 levels of counseling, into HIV clinical care. METHODS: HIV-positive, opioid-dependent patients were enrolled in a 12-week pilot study and randomized to receive daily buprenorphine/naloxone treatment along with either brief physician management or physician management combined with nurse administered drug counseling and adherence management. Primary outcomes included treatment retention; illicit drug use, assessed by urine toxicology test and self report; CD4 lymphocyte counts; and log(10) HIV type 1 (HIV-1) RNA levels. RESULTS: Of the 16 patients who received at least 1 dose of buprenorphine, 13 (81%) completed 12 weeks of treatment. The proportion of opioid-positive weekly urine test results decreased from 100% at baseline to 32% (month 1), 20% (month 2), and 16% (month 3). Only 4 patients reported any opioid use (in the prior 7 days) during the 12-week study. CD4 lymphocyte counts remained stable over the course of the study. The mean log(10) HIV-1 RNA level (+/- standard deviation) declined significantly, from 3.66+/-1.06 log(10) HIV-1 RNA copies/mL at baseline to 3.0+/-0.57 log(10) HIV-1 RNA copies/mL at month 3 (P<.05). No significant differences based on counseling intervention were detected. All 13 patients who completed the study continued to receive treatment in an extension phase of at least 0-15 months' duration. CONCLUSIONS: We conclude that it is feasible to integrate buprenorphine into HIV clinical care for the treatment of opioid dependence. Patients experienced good treatment retention and reductions in their opioid use. HIV biological markers remained stable or improved during buprenorphine/naloxone treatment. PMID- 17109306 TI - Initial strategies for integrating buprenorphine into HIV care settings in the United States. AB - The Centers for Disease Control and Prevention's HIV Prevention Strategic Plan Through 2005 advocated for increasing the proportion of persons with human immunodeficiency virus (HIV) infection and in need of substance abuse treatment who are successfully linked to services for these 2 conditions. There is evidence that integrating care for HIV infection and substance abuse optimizes outcomes for patients with both disorders. Buprenorphine, a recently approved medication for the treatment of opioid dependence in physicians' offices, provides the opportunity to integrate the treatment of HIV infection and substance abuse in one clinical setting, yet little information exists on the models of care that will most successfully facilitate this integration. To promote the uptake of this type of integrated care, the current review provides a description of 4 recently implemented models for combining buprenorphine treatment with HIV primary care: (1) an on-site addiction/HIV specialist treatment model; (2) a HIV primary care physician model; (3) a nonphysician health professional model; and (4) a community outreach model. PMID- 17109307 TI - Buprenorphine use: the international experience. AB - The confluence of the heroin injection epidemic and the human immunodeficiency virus (HIV) infection epidemic has increased the call for expanded access to effective treatments for both conditions. Buprenorphine and methadone are now listed on the World Health Organization's Model Essential Drugs List. In France, which has the most extensive experience, buprenorphine has been associated with a dramatic decrease in deaths due to overdose, and buprenorphine diversion appears to be associated with inadequate dosage, social vulnerability, and prescriptions from multiple providers. Other treatment models (in the United States, Australia, Germany, and Italy) and buprenorphine use in specific populations are also reviewed in the present article. In countries experiencing a dual epidemic of heroin use and HIV infection, such as former states of the Soviet Union and other eastern European and Asian countries, access to buprenorphine and methadone may be one potential tool for reducing the spread of HIV infection among injection drug users and for better engaging them in medical care. PMID- 17109308 TI - Pharmacokinetic interactions between buprenorphine and antiretroviral medications. AB - Buprenorphine is used for the treatment of opioid dependence. As the number of persons receiving buprenorphine treatment and antiretroviral therapy continues to grow, so too does the existence and clinical impact of drug interactions between buprenorphine and medications for treating human immunodeficiency virus (HIV) infection. Awareness that such interactions exist may deter some patients and physicians from initiating potentially lifesaving therapy or lead to complications among patients whose treatment is already under way. Complications include nonadherence to antiretroviral therapy and the development of viral resistance. Illicit drug use is a frequent consequence of adverse drug effects experienced by injection drug users. The occurrence of unrecognized drug interactions can lead to unsuccessful therapy for HIV infection and the treatment of substance dependence. The present review is organized to provide a working background of buprenorphine pharmacology. Review of the current state of knowledge regarding specific interactions between buprenorphine and antiretrovirals is followed by a review of the clinical applicability of these interactions. PMID- 17109309 TI - Interactions between buprenorphine and antiretrovirals. I. The nonnucleoside reverse-transcriptase inhibitors efavirenz and delavirdine. AB - This study examined drug interactions between buprenorphine, an opioid partial agonist medication used in the treatment of opioid dependence, and the nonnucleoside reverse-transcriptase inhibitors (NNRTIs) efavirenz (EFV) and delavirdine (DLV). Opioid-dependent, buprenorphine/naloxone-maintained, human immunodeficiency virus (HIV)-negative volunteers (n=10 per NNRTI) participated in 24-h sessions to determine pharmacokinetics of buprenorphine and of buprenorphine with either EFV or DLV after administration of standard doses of either antiretroviral for 15 or 7 days, respectively. Opiate withdrawal symptoms, cognitive effects, and adverse events were determined before and after antiretroviral administration in opioid-dependent participants. The pharmacokinetics of NNRTIs in healthy control participants were used to determine the effect of buprenorphine on NNRTIs. EFV decreased the buprenorphine area under the concentration-time curve (P<.001). DLV increased buprenorphine concentrations (P<.001). Clinically significant consequences of these interactions were not observed. Buprenorphine did not alter antiretroviral pharmacokinetics. Adjustments of doses of either buprenorphine or EFV or DLV are not likely to be necessary when these drugs are administered for the treatment of opiate dependence and HIV disease. PMID- 17109310 TI - Interactions between buprenorphine and antiretrovirals. II. The protease inhibitors nelfinavir, lopinavir/ritonavir, and ritonavir. AB - We examined drug interactions between buprenorphine, an opioid partial agonist available by prescription for treatment of opioid dependence, and the protease inhibitors (PIs) nelfinavir (NFV), ritonavir (RTV), and lopinavir/ritonavir (LPV/R). Opioid-dependent, buprenorphine/naloxone-maintained, human immunodeficiency virus (HIV)-negative volunteers (n=10 per PI) participated in 24 h pharmacokinetic studies, before and after administration of each PI. Symptoms of opiate withdrawal and excess were determined before and after PI administration. PI pharmacokinetics were determined and compared between opiate dependent participants and healthy control participants (n=15 per PI). Administration of RTV, but not of NFV or LPV/R, resulted in a significant increase in the buprenorphine area under the concentration-time curve (AUC). Symptoms of opiate excess, however, were not observed. Buprenorphine had no significant effects on PI AUC. Adjustments of doses of either buprenorphine or NFV, LPV/R, or RTV are not likely to be necessary when these drugs are administered for the treatment of opioid dependence and HIV disease. PMID- 17109311 TI - Overcoming policy and financing barriers to integrated buprenorphine and HIV primary care. AB - Treatment for substance abuse and human immunodeficiency virus (HIV) infection historically have come from different providers, often in separate locations, and have been reimbursed through separate funding streams. We describe policy and financing challenges faced by health care providers seeking to integrate buprenorphine, a new treatment for opioid dependence, into HIV primary care. Regulatory challenges include licensing and training restrictions imposed by the Drug Addiction Treatment Act of 2000 and confidentiality regulations for alcohol and drug treatment records. Potential responses include the development of local training programs and electronic medical records. Addressing the complexity of funding sources for integrated care will require administrative support, up-front investments, and federal and state leadership. A policy and financing research agenda should address evidence gaps in the rationales for regulatory restrictions and should include cost-effectiveness studies that quantify the "value for money" of investments in integrated care to improve health outcomes for HIV-infected patients with opioid dependence. PMID- 17109312 TI - Buprenorphine and HIV primary care: report of a forum for collaborative HIV research workshop. AB - On 3-4 June 2004, in Washington, DC, the Forum for Collaborative HIV Research convened experts from academia, community and private practices, US government agencies, and industry to develop recommendations for increased uptake of buprenorphine integrated into human immunodeficiency virus (HIV) primary care, with special emphasis on Ryan White CARE Act-funded programs. Workshop participants evaluated knowledge gaps requiring research; barriers to integration at the patient, clinic, and systems level; policy and financing issues; and program impacts. Recommendations were developed for training, including medical school and post-medical school training of clinical teams as well as training of patients; for improving programs and services, including integration of opioid dependence and HIV infection into chronic disease models, providing flexible access to core and support services, and monitoring and evaluation of programs; for changes in policy supportive of program and services goals; for financing buprenorphine treatment by use of existing models of integrated treatment and merging funding streams at the local level; and for addressing research gaps, including cost-effectiveness research. PMID- 17109313 TI - Comparative analysis of worker reproduction and policing in eusocial hymenoptera supports relatedness theory. AB - In many bees, wasps, and ants, workers police each other in order to prevent individual workers from selfishly producing their own male offspring. Although several factors can selectively favor worker policing, genetic relatedness is considered to be of special importance. In particular, kin selection theory predicts that worker policing should be more common in species where workers are more related to the queen's sons than to other workers' sons. Here we provide strong novel support for this theory based on a comparative analysis of policing and male parentage in 109 species of ants, bees, and wasps. First, an analysis of behavioral data confirms that worker policing occurs more frequently in species where workers are more related to the queen's sons than to other workers' sons. Second, an analysis of male parentage shows that a significantly higher percentage of the males are workers' sons in species where the workers are more related to other workers' sons. Both conclusions also hold if data are analyzed using phylogenetically independent contrasts. Although our analysis provides strong overall support for the theory that relatedness affects kin conflict over male parentage, there is also significant residual variation. Several factors that may explain this variation are discussed. PMID- 17109314 TI - Why animals lie: how dishonesty and belief can coexist in a signaling system. AB - We develop and apply a simple model for animal communication in which signalers can use a nontrivial frequency of deception without causing listeners to completely lose belief. This common feature of animal communication has been difficult to explain as a stable adaptive outcome of the options and payoffs intrinsic to signaling interactions. Our theory is based on two realistic assumptions. (1) Signals are "overheard" by several listeners or listener types with different payoffs. The signaler may then benefit from using incomplete honesty to elicit different responses from different listener types, such as attracting potential mates while simultaneously deterring competitors. (2) Signaler and listener strategies change dynamically in response to current payoffs for different behaviors. The dynamic equations can be interpreted as describing learning and behavior change by individuals or evolution across generations. We explain how our dynamic model differs from other solution concepts from classical and evolutionary game theory and how it relates to general models for frequency-dependent phenotype dynamics. We illustrate the theory with several applications where deceptive signaling occurs readily in our framework, including bluffing competitors for potential mates or territories. We suggest future theoretical directions to make the models more general and propose some possible experimental tests. PMID- 17109315 TI - Pupal remodeling and the development and evolution of sexual dimorphism in horned beetles. AB - Horns or hornlike structures in beetles have become an increasingly popular study system for exploring the evolution and development of secondary sexual trait diversity and sexual dimorphisms. The horns of adult beetles originate during a rapid growth phase during the prepupal stage of larval development, and differential activation of growth during this time is either implicitly or explicitly assumed to be the sole mechanism underlying intra- and interspecific differences in adult horn expression. Here I show that this assumption is not based on developmental reality. Instead, after their initial prepupal growth phase, beetle horns are extensively remodeled during the subsequent pupal stage via sex- and size-dependent resorption of horn tissue. I show that adult sexual dimorphism in four Onthophagus species is shaped partly or entirely by such pupal remodeling rather than by differential growth. Specifically, I show that after a sexually monomorphic growth phase, differential pupal horn resorption can generate both regular and reversed sexual dimorphism. Furthermore, I show that in cases in which initial growth is already dimorphic, pupal horn resorption can both magnify and reverse initial dimorphism resulting from differential growth. Finally, I show that complete resorption of pupal horns in both sexes can remove any trace of horn expression from all resulting adults. In such species, examination of adults only would result in the false conclusion that this species lacks the ability to develop a horn. Instead, such species appear to differ from those with sexually dimorphic adults merely in that they activate pupal horn resorption in both sexes rather than in just one. Combined, these results suggest that pupal remodeling of secondary trait expression is taxonomically widespread, at least among Onthophagus species, and is developmentally extensive and remarkably evolutionarily labile. These results have immediate implications for reconstructing the evolutionary history of horned beetles and the role of developmental processes in guiding evolutionary trajectories. I use these results to revise current understanding of the evolutionary developmental biology of secondary sexual traits in horned beetles in particular and holometabolous insects in general. The results presented here seriously call into question whether descriptions of adult diversity patterns alone suffice for meaningful inferences toward understanding the developmental and evolutionary origin of these patterns. These results illustrate that a lasting integration of development into an evolutionary framework must integrate development as a process rather than define it solely by some of its products. PMID- 17109316 TI - Vicariant origin of malagasy reptiles supports late cretaceous antarctic land bridge. AB - Since the acceptance of Wegener's theory of plate tectonics in the 1960s, continental drift vicariance has been proposed as an explanation for pan Gondwanan faunal distributions. Given the recognition of historical connections among continents, it no longer was necessary to invoke hypotheses of dispersal across nearly insurmountable barriers. The application of continental drift vicariance theory to Gondwanan floral and faunal distributions provided reasonable explanations for such unusual distributions as that of the southern beech (Nothofagus) and chameleons. However, recent studies have demonstrated a significant, if not dominant, role for dispersal in the present-day distributions of these and numerous other "Gondwanan" taxa. The evolutionary histories of three Malagasy groups (boid snakes, podocnemid turtles, and iguanid lizards) commonly have been interpreted as reflecting vicariance because of continental drift associated with the breakup of Gondwana. Bayesian analyses of divergence ages suggest that this pattern is the result of vicariance coincident with the isolation of Madagascar in the Late Cretaceous (approximately 80 million years ago). This represents the first temporal evidence linking the vicariant origin of extant Malagasy vertebrates to a single geologic event. Specifically, our data provide strong, independently corroborated evidence for a contiguous Late Cretaceous Gondwana, exclusive of Africa and connected via Antarctica. PMID- 17109317 TI - Sexual conflict and the evolution of female preferences for indicators of male quality. AB - Males and females have opposing interests when it comes to the honesty of signals used in mate choice. The existence of this sexual conflict has long been acknowledged, but its consequences have not been fully investigated. By applying adaptive dynamics methods and individual-based computer simulations to a standard model for good-genes sexual selection, we show that sexual conflict over condition-dependent signaling can prevent the handicap process from ever attaining an evolutionary equilibrium. We outline the parameter conditions and properties of the underlying genetics conducive to nonequilibrium behavior and discuss the potential of such behavior to explain the elaboration and frequent phylogenetic loss of sexually selected traits. We also evaluate its consequences for well-established insights of sexual selection theory previously shown to apply when female mating preference and male ornament expression do converge on stable equilibrium levels. Contrary to equilibrium expectation, a continual change of condition-dependent signaling enables the evolution of a costly preference for a pure epistatic indicator and the evolution of preferences for redundant signals or a large number of independent ornaments. We thus conclude that seemingly general results of sexual selection theory, insofar as these are based on equilibrium considerations, do not extend to cases where nonequilibrium behavior occurs. PMID- 17109318 TI - Manipulating lek size and composition using decoys: an experimental investigation of lek evolution models. AB - Four theoretical models have been proposed to account for the origin and maintenance of leks: hotspot, female preference, hotshot, and black hole models. Each has been validated in particular cases, and most are not mutually exclusive; therefore, it has been difficult to contrast and separate them, empirically and experimentally. By using decoys to mimic natural leks in the little bustard, artificial leks attracted wild birds. Then, by manipulating artificial lek size and structure (sex ratio, male phenotype), the study of responses of wild males and females allowed us to test specific predictions derived from the four classical models of lek evolution. The hotspot model was not supported because female decoys did not attract wild males. Conversely, hotshot males do exist in this species (attracting both wild females and males), as does a female preference for a particular lek size (four males). Finally, males aggressive toward decoys attracted fewer females, consistent with one of the mechanisms by which the black hole model may work. Therefore, three models of lek evolution were partly or fully supported by our experimental results: hotshot, female preference, and black hole models. We suggest that these models actually fit within each other, ensuring the evolution, functioning, and long-term maintenance of leks. PMID- 17109319 TI - Metapopulation dynamics and biological invasions: a spatially explicit model applied to a freshwater snail. AB - The spatial spread of invading organisms is a major contemporary concern. We focus here on invasions in inherently fragmented habitats, such as freshwater systems, and explore the usefulness of metapopulation models in this context. Maximum-likelihood methods allow the estimation of colonization and extinction rates, as functions of habitat patch sizes and positions, from time series of presence/absence data. This framework also provides confidence intervals of these estimates and hypotheses tests. We analyze a previously unpublished 12-year survey of the spread of the introduced snail Tarebia granifera in 47 Martinican rivers. Simple metapopulation models reproduce with reasonable accuracy several quantitative aspects of the invasion, including regional abundance, spatiotemporal structure, and site-by-site colonization dates. Sensitivity analysis reveals that the invasion sequence depended strongly on metapopulation size (number of sites) and spatial structure (distances among sites). The invasion history has also been accelerated by stochastic events, as illustrated by a large, central river that happened to be colonized very early and served as an invasion pool. Finally, we discuss the benefits of this approach for the understanding of invasions in fragmented landscapes. PMID- 17109320 TI - Deciphering the effects of climate on animal populations: diagnostic analysis provides new interpretation of Soay sheep dynamics. AB - Soay sheep on the island of Hirta exhibit periodic population collapses that have been proposed to result from nonlinear interactions between weather, population density, and age structure. Here we employ a diagnostic approach to reanalyze the data from 1985 to 2004 and find that climate mainly affects the equilibrium population size, thus acting as a lateral perturbation. From this, we derive a simple energetic model for a population interacting with its food supply in the presence of variable winter weather. This model explains the strong nonlinearity in the Soay sheep population regulation function and provides a framework for evaluating climatic perturbations. We examined two integrative climatic indexes, one representing effects on forage production and the other representing the severity of winter weather. Results suggest that the latter has the main effect on Soay sheep population dynamics. Models incorporating this variable provided fairly accurate predictions of Soay sheep population fluctuations. The diagnostic approach offers an objective way to develop simple, nonstructured population models that are useful for understanding the causes of population fluctuations and predicting population changes, provided they are based on a careful consideration of the underlying biological and/or ecological mechanisms. PMID- 17109321 TI - Evidence for a time-integrated species-area effect on the latitudinal gradient in tree diversity. AB - The greater area of tropical forest biomes has been proposed as a factor that drives the latitudinal gradient in species diversity by modulating speciation and extinction rates. But speciation and extinction are processes that operate over millions of years, so an adequate test of area's contribution to diversity patterns must take into consideration that biome areas have changed through time in response to climate. Here we correlate estimates of current tree species diversity with a composite parameter integrating area over geological time for each continent's tropical, temperate, and boreal biomes. We find significant positive correlations between current tree diversity and area-time for periods since the Eocene, Oligocene, and Miocene, which we take as evidence for a time integrated species-area effect on current patterns of species richness across biomes. These results contribute to explanations for why most lineages have tropical origins and why tropical forests are more diverse than extratropical forests. PMID- 17109322 TI - Rate of lineage origin explains the diversity anomaly in the world's mangrove vegetation. AB - The contribution of nonecological factors to global patterns in diversity is evident when species richness differs between regions with similar habitats and geographic area. Mangrove environments in the Eastern Hemisphere harbor six times as many species of trees and shrubs as similar environments in the New World. Genetic divergence of mangrove lineages from terrestrial relatives, in combination with fossil evidence, suggests that mangrove diversity is limited by evolutionary transition into the stressful marine environment, the number of mangrove lineages has increased steadily over the Tertiary with little global extinction, and the diversity anomaly in mangrove vegetation reflects regional differences in the rate of origin of new mangrove lineages. PMID- 17109323 TI - Habitat selection responses of parents to offspring predation risk: an experimental test. AB - The ability of nest predation to influence habitat settlement decisions in birds is widely debated, despite its importance in limiting fitness. Here, we experimentally manipulated nest predation risk across a landscape and asked the question, do migratory birds assess and respond to variation in nest predation risk when choosing breeding habitats? We examined habitat preference by quantifying the density and settlement date of eight species of migratory passerines breeding in areas with and without intact nest predator communities. We found consistently more individuals nesting in areas with reduced nest predation than in areas with intact predator assemblages, although predation risk had no influence on settlement or breeding phenology. Additionally, those individuals occupying safer nesting habitats exhibited increased singing activity. These findings support a causal relationship between habitat choice and nest predation risk and suggest the importance of nest predation risk in shaping avian community structure and breeding activity. PMID- 17109324 TI - What hypotheses are you willing to entertain? AB - The 2005 American Society of Naturalists Vice-Presidential Symposium was organized on the theme of "Integrating across Scientific Disciplines." Integration implies considering some aspect of nature from disparate vantages that cut across disciplinary boundaries. The articles presented here illustrate that taking integrative approaches to addressing research questions yields much greater insights than do piecemeal contributions by separate fields. PMID- 17109325 TI - The sorry state of F2 hybrids: consequences of rapid mitochondrial DNA evolution in allopatric populations. AB - Through the processes of natural selection and genetic drift, allopatric populations diverge genetically and may ultimately become reproductively incompatible. In cases of prezygotic reproductive isolation, candidate systems for speciation genes logically include genes involved in mate or gamete recognition. However, where only postzygotic isolation exists, candidate speciation genes could include any genes that affect hybrid performance. We hypothesize that because mitochondrial genes frequently evolve more rapidly than the nuclear genes with which they interact, interpopulation hybridization might be particularly disruptive to mitochondrial function. Understanding the potential impact of intergenomic (nuclear and mitochondrial) coadaptation on the evolution of allopatric populations of the intertidal copepod Tigriopus californicus has required a broadly integrative research program; here we present the results of experiments spanning the spectrum of biological organization in order to demonstrate the consequences of molecular evolution on physiological performance and organismal fitness. We suggest that disruption of mitochondrial function, known to result in a diverse set of human diseases, may frequently underlie reduced fitness in interpopulation and interspecies hybrids in animals. PMID- 17109326 TI - Biological stoichiometry: a chemical bridge between ecosystem ecology and evolutionary biology. AB - The mission of the American Society of Naturalists is "to advance and diffuse knowledge of organic evolution and other broad biological principles so as to enhance the conceptual unification of the biological sciences." In this article, I argue that the area of biology least integrated with knowledge of organic evolution is the field of ecosystem ecology, as evidenced by a semiquantitative literature survey of use of terms in the scientific literature. I present an overview of recent theoretical developments and empirical findings in the emerging field of biological stoichiometry (the study of the balance of energy and multiple chemical elements in living systems). These developments hold some promise as a means to conceptually integrate ecosystem ecology, with its emphasis on flows and pools of energy and chemical elements, with evolutionary biology, with its emphasis on genetic fitness and the biochemical products of the genome. For example, recent evidence indicates that organismal C : P and N : P ratios have a major impact on biologically mediated flows of energy and phosphorus; in turn, variations among taxa in these ratios are connected to evolved differences in organismal growth rate because of the connection between growth rate and the need for increased allocation to P-rich ribosomal RNA. In this way, evolutionary change in growth-related traits, by altering organismal P requirements, has direct biogeochemical implications, while ecosystem conditions can constrain evolutionary acceleration of growth rates by imposing a direct P limitation on production of the needed biochemical machinery of growth. Thus, stoichiometric theory provides a broad biological principle that can interconvert the currencies and concerns of ecosystem ecology and evolutionary biology, facilitating integration of diverse fields of study and contributing to conceptual unification of the biological sciences. PMID- 17109327 TI - Directional changes in ecological communities and social-ecological systems: a framework for prediction based on Alaskan examples. AB - In this article we extend the theory of community prediction by presenting seven hypotheses for predicting community structure in a directionally changing world. The first three address well-studied community responses to environmental and ecological change: ecological communities are most likely to exhibit threshold changes in structure when perturbations cause large changes in limiting soil or sediment resources, dominant or keystone species, or attributes of disturbance regime that influence community recruitment. Four additional hypotheses address social-ecological interactions and apply to both ecological communities and social-ecological systems. Human responsiveness to short-term and local costs and benefits often leads to human actions with unintended long-term impacts, particularly those that are far from the site of decision making or are geographically dispersed. Policies are usually based on past conditions of ecosystem services rather than expected future trends. Finally, institutions that strengthen negative feedbacks between human actions and social-ecological consequences can reduce human impacts through more responsive (and thus more effective) management of public ecosystem services. Because of the large role that humans play in modifying ecosystems and ecosystem services, it is particularly important to test and improve social-ecological hypotheses as a basis for shaping appropriate policies for long-term ecosystem resilience. PMID- 17109328 TI - Exploring evolutionary constraints is a task for an integrative evolutionary biology. AB - Judging by the volume of writings about evolutionary constraints, they are an important topic in evolutionary biology. However, their involvement in shaping patterns of evolutionary change from morphological stasis to adaptive radiation remains contentious. This is at least in part because of the paucity of robust analyses of potential examples of constraints, whether of a more absolute or a relative nature. Here, we argue that what is needed to explore the type of constraints and bias on evolutionary change that may emerge from the way in which phenotypic variation is generated is an integrative approach applied to systems that can be tackled at different levels of biological organization. This is illustrated using research on the evolution of patterns in butterfly wing eyespots that has applied a combination of evolutionary genetics and evo-devo to an emerging model species with the beginnings of a comparative approach to describe patterns of variability among the extant taxa of two species-rich genera. PMID- 17109329 TI - A tale of two diversifications: reciprocal habitat shifts to fill ecological space along the pond permanence gradient. AB - The Enallagma and Lestes damselflies have both diversified and adapted over the past 10-15 million years to the various ecological milieus found along the pond permanence gradient among North American ponds and lakes. Previous articles have explored this diversification process for Enallagma. In this article, we present a phylogenetic hypothesis for the North American Lestes, use this hypothesis to reconstruct Lestes diversification, and compare the diversification processes inferred for Lestes and Enallagma. The results of this study suggest that Lestes began in temporary ponds where large dragonflies are the top predators, while Enallagma began in permanent lakes where fish are the top predators. Starting from these different ancestral habitats, both genera have invaded and adapted to habitats already occupied by the other genus. Moreover, these adaptive habitat shifts involved substantial convergence on the behaviors used to deal with fish and dragonfly predation in both genera and a major life-history shift from diapausing to directly developing eggs in Lestes. However, in Lestes lineages invading fish lakes, swimming speed and morphology did not change to match those of Enallagma species, illustrating that reciprocal shifts between alternative selection regimes are not necessarily evolutionary opposites. Also, the greater sizes and growth rates of Lestes species compared to Enallagma species, which should impart substantial ecological advantages in competition between the genera, were shown to result from phylogenetic inheritance and not from adaptive diversification. This historical analysis of diversification raises new questions about the relationship between the macroevolutionary mechanisms driving lineage diversification and the ecological mechanisms structuring local food webs and regional species assemblages. PMID- 17109330 TI - Integration without unification: an argument for pluralism in the biological sciences. AB - In this article, we consider the tension between unification and pluralism in biological theory. We begin with a consideration of historical efforts to establish a unified understanding of evolution in the neo-Darwinian synthesis. The fragmentation of the evolutionary synthesis by molecular evolution suggests the limitations of the general unificationist ideal for biology but not necessarily for integrating explanations. In the second half of this article, we defend a specific variety of pluralism that allows for the integration required for explanations of complex phenomena without unification on a large scale. PMID- 17109331 TI - Phase 1 clinical trials of the National Institutes of Health Vaccine Research Center HIV/AIDS candidate vaccines. PMID- 17109332 TI - Is the glass three-quarters full or one-quarter empty? PMID- 17109333 TI - Immune-based therapy for HIV infection: are acute and chronic HIV infection different diseases? PMID- 17109334 TI - Finding the cause of Kawasaki disease: a pediatric infectious diseases research priority. PMID- 17109335 TI - Phase 1 safety and immunogenicity evaluation of a multiclade HIV-1 candidate vaccine delivered by a replication-defective recombinant adenovirus vector. AB - BACKGROUND: The development of an effective human immunodeficiency virus (HIV) vaccine is a high global priority. Here, we report the safety, tolerability, and immunogenicity of a replication-defective recombinant adenovirus serotype 5 (rAd5) vector HIV-1 candidate vaccine. METHODS: The vaccine is a mixture of 4 rAd5 vectors that express HIV-1 subtype B Gag-Pol fusion protein and envelope (Env) from subtypes A, B, and C. Healthy, uninfected adults were randomized to receive 1 intramuscular injection of placebo (n=6) or vaccine at dose levels of 10(9) (n=10), 10(10) (n=10), or 10(11) (n=10) particle units and were followed for 24 weeks to assess immunogenicity and safety. RESULTS: The vaccine was well tolerated but was associated with more reactogenicity at the highest dose. At week 4, vaccine antigen-specific T cell responses were detected in 28 (93.3%) and 18 (60%) of 30 vaccine recipients for CD4(+) and CD8(+) T cells, respectively, by intracellular cytokine staining assay and in 22 (73%) of 30 vaccine recipients by enzyme-linked immunospot assay. Env-specific antibody responses were detected in 15 (50%) of 30 vaccine recipients by enzyme-linked immunosorbant assay and in 28 (93.3%) of 30 vaccine recipients by immunoprecipitation followed by Western blotting. No neutralizing antibody was detected. CONCLUSIONS: A single injection induced HIV-1 antigen-specific CD4(+) T cell, CD8(+) T cell, and antibody responses in the majority of vaccine recipients. This multiclade rAd5 HIV-1 vaccine is now being evaluated in combination with a multiclade HIV-1 DNA plasmid vaccine. PMID- 17109336 TI - Phase 1 safety and immunogenicity evaluation of a multiclade HIV-1 DNA candidate vaccine. AB - BACKGROUND: Gene-based vaccine delivery is an important strategy in the development of a preventive vaccine for acquired immunodeficiency syndrome (AIDS). Vaccine Research Center (VRC) 004 is the first phase 1 dose-escalation study of a multiclade HIV-1 DNA vaccine. METHODS: VRC-HIVDNA009-00-VP is a 4 plasmid mixture encoding subtype B Gag-Pol-Nef fusion protein and modified envelope (Env) constructs from subtypes A, B, and C. Fifty healthy, uninfected adults were randomized to receive either placebo (n=10) or study vaccine at 2 mg (n=5), 4 mg (n=20), or 8 mg (n=15) by needle-free intramuscular injection. Humoral responses (measured by enzyme-linked immunosorbant assay, Western blotting, and neutralization assay) and T cell responses (measured by enzyme linked immunospot assay and intracellular cytokine staining after stimulation with antigen-specific peptide pools) were measured. RESULTS: The vaccine was well tolerated and induced cellular and humoral responses. The maximal CD4(+) and CD8(+) T cell responses occurred after 3 injections and were in response to Env peptide pools. The pattern of cytokine expression by vaccine-induced HIV-specific T cells evolved over time, with a diminished frequency of interferon- gamma producing T cells and an increased frequency of interleukin-2-producing T cells at 1 year. CONCLUSIONS: DNA vaccination induced antibody to and T cell responses against 3 major HIV-1 subtypes and will be further evaluated as a potential component of a preventive AIDS vaccine regimen. PMID- 17109337 TI - Randomized, double-blind, placebo-controlled efficacy trial of a bivalent recombinant glycoprotein 120 HIV-1 vaccine among injection drug users in Bangkok, Thailand. AB - BACKGROUND: In Thailand, phase 1/2 trials of monovalent subtype B and bivalent subtype B/E (CRF01_AE) recombinant glycoprotein 120 human immunodeficiency virus type 1 (HIV-1) vaccines were successfully conducted from 1995 to 1998, prompting the first HIV-1 vaccine efficacy trial in Asia. METHODS: This randomized, double blind, placebo-controlled efficacy trial of AIDSVAX B/E (VaxGen), which included 36-months of follow-up, was conducted among injection drug users (IDUs) in Bangkok, Thailand. The primary end point was HIV-1 infection; secondary end points included plasma HIV-1 load, CD4 cell count, onset of acquired immunodeficiency syndrome-defining conditions, and initiation of antiretroviral therapy. RESULTS: A total of 2546 IDUs were enrolled between March 1999 and August 2000; the median age was 26 years, and 93.4% were men. The overall HIV-1 incidence was 3.4 infections/100 person-years (95% confidence interval [CI], 3.0 3.9 infections/100 person-years), and the cumulative incidence was 8.4%. There were no differences between the vaccine and placebo arms. HIV-1 subtype E (83 vaccine and 81 placebo recipients) accounted for 77% of infections. Vaccine efficacy was estimated at 0.1% (95% CI, -30.8% to 23.8%; P=.99, log-rank test). No statistically significant effects of the vaccine on secondary end points were observed. CONCLUSION: Despite the successful completion of this efficacy trial, the vaccine did not prevent HIV-1 infection or delay HIV-1 disease progression. PMID- 17109338 TI - A randomized, partially blinded phase 2 trial of antiretroviral therapy, HIV specific immunizations, and interleukin-2 cycles to promote efficient control of viral replication (ACTG A5024). AB - Strategies to limit life-long dependence on antiretroviral therapy (ART) are needed. We randomized 81 human immunodeficiency virus (HIV)-infected subjects to 4 interventional arms involving continued ART plus ALVAC vCP1452 (or placebo) with or without interleukin (IL)-2 infusions. Viral load rebound 12 weeks after ART interruption was then analyzed to assess immune control. Fifty-two subjects reached the study end point. ALVAC recipients had 0.5 log(10) lower virologic rebounds (P=.033). IL-2 plus vaccine boosted CD4(+) T cell counts (P<.001) but did not diminish viral rebound. Significant changes were not detected for HIV specific lymphoproliferative responses in any arm. This exploratory protocol provides useful clinical data for future therapeutic immunization trial design. PMID- 17109339 TI - Cyclosporin A provides no sustained immunologic benefit to persons with chronic HIV-1 infection starting suppressive antiretroviral therapy: results of a randomized, controlled trial of the AIDS Clinical Trials Group A5138. AB - BACKGROUND: Although the determinants of immune deficiency and immune restoration in chronic human immunodeficiency virus (HIV)-1 infection are not well understood, immune activation has been proposed as being central to the pathogenesis of HIV. METHODS: A randomized, controlled trial of cyclosporin A treatment for 2 weeks was performed in persons with chronic HIV-1 infection who were beginning a standardized antiretroviral therapy (ART) regimen. RESULTS: Treatment with cyclosporin A provided only a marginal and transient enhancement in circulating T cell restoration that was largely restricted to cells expressing the CCR7 chemokine receptor and that did not persist beyond 2 weeks. CONCLUSIONS: Cyclosporin A coadministered for 2 weeks with ART provided no sustained immunologic benefit to persons with chronic HIV-1 infection. If immune activation drives progressive immune deficiency in chronic HIV-1 infection, these activation pathways may not be sensitive to cyclosporin. PMID- 17109340 TI - Treatment benefit on cerebrospinal fluid HIV-1 levels in the setting of systemic virological suppression and failure. AB - OBJECTIVE: To characterize the effect of partially suppressive combination antiretroviral therapy on cerebrospinal fluid (CSF) human immunodeficiency virus (HIV)-1 RNA levels and CSF inflammation. DESIGN: The study was a cross-sectional analysis of 139 HIV-1-infected subjects without active neurological disease, categorized as having successful therapy (plasma HIV-1 RNA level < or =500 copies/mL), having failure of therapy (plasma HIV-1 RNA level >500 copies/mL), or not receiving therapy. The control group consisted of 48 HIV-negative subjects. CSF and plasma HIV-1 RNA assays had a lower limit of quantification of 2.5 copies/mL. Genotypic resistance testing was performed on a subset of subjects. RESULTS: Of the 47 subjects with successful therapy, CSF HIV-1 RNA levels were <2.5 copies/mL in 34 (72%). Only 1 had an HIV-1 RNA level >500 copies/mL. Although plasma HIV-1 RNA levels were similar in 35 subjects with failed therapy and 57 of those not receiving therapy (P=.84), CSF HIV-1 RNA levels were at least 10-fold lower in subjects with failed therapy (P<.0001). This disproportionate effect of treatment on CSF HIV-1 RNA levels was found across the range of plasma HIV-1 RNA levels and was not explained by differences in levels of drug resistance in plasma or CSF. Therapy reduced CSF inflammation in both treated groups. CONCLUSIONS: In our cohort, antiretroviral therapy had a greater effect on HIV-1 RNA levels in CSF than in plasma and reduced intrathecal inflammation, even in the presence of drug resistance. PMID- 17109341 TI - Blinded case-control study of the relationship between human coronavirus NL63 and Kawasaki syndrome. AB - We conducted a blinded, case-control, retrospective study in pediatric patients hospitalized at The Children's Hospital, Denver, Colorado, to determine whether human coronavirus (HCoV)-NL63 infection is associated with Kawasaki syndrome (KS). Over the course of a 7-month period, nasopharyngeal-wash samples from 2 (7.7%) of 26 consecutive children with KS and 4 (7.7%) of 52 matched control subjects tested positive for HCoV-NL63 by reverse transcription-polymerase chain reaction. These data suggest that, although HCoV-NL63 was circulating in children in our community during the time of the study, the prevalence of infection with HCoV-NL63 was not greater in patients with KS than in control subjects. PMID- 17109342 TI - Human papillomavirus prevalence in women who have and have not undergone hysterectomies. AB - We compared human papillomavirus (HPV) prevalence in an age-stratified random sample of women who have undergone a hysterectomy (WH) (n=573) with the HPV prevalence in age-matched women with intact cervices (women who have not undergone a hysterectomy [WNH]) (n=581) participating in a study at Kaiser Permanente in Portland, Oregon. Testing cervicovaginal lavage fluids for >40 HPV genotypes using an MY09/11 L1 consensus primer polymerase chain reaction method, we found no statistical differences in the prevalence of HPV (16% for WNH vs. 13.9% for WH) or carcinogenic HPV (6.5% for WNH vs. 4.5% for WH) between the 2 groups of women. Although WH have a similar prevalence of carcinogenic HPV infection, compared with WNH without a cervix, they have minimal risk of HPV induced cancer and are unlikely to benefit from HPV testing. PMID- 17109343 TI - Viral load, E2 gene disruption status, and lineage of human papillomavirus type 16 infection in cervical neoplasia. AB - The clinical utility of human papillomavirus (HPV) load and integration status remains unclear. We applied refined methods to delineate the viral load, integration status, and lineage of 104 women with HPV-16 monotype infection, including 19 with normal cervices, 9 with histologically proven cervical intraepithelial neoplasia (CIN) 1, 24 with CIN 2, 27 with CIN 3, and 25 with squamous cell carcinoma (SCC). Higher crude viral load, as determined by real time polymerase chain reaction (PCR) targeting the E7 gene, was observed for SCC but became insignificant after normalization for cell content. Integration was located and quantified by real-time PCRs targeting, respectively, the carboxyl, amino, and hinge domains of the E2 gene. Pure episomal, integrated, and mixed forms were observed in all disease groups. Most E2 gene disruptions involved the amino-terminal, but sparing the hinge region that has been frequently used as a surrogate marker of integration. Large-fragment disruption involving all 3 E2 regions was observed only in the CIN 3 and SCC groups. Altogether, 33.3% of the CIN 3 group and 28.0% of the SCC group harbored pure episomal genomes. The Asian lineage was associated with a higher risk for CIN 3/SCC than the European lineage, and 6 of the 7 large-fragment E2 disruptions were from Asian lineage. The link between viral lineage, integration pattern, and oncogenesis deserves further study. PMID- 17109344 TI - Equal amounts of intracellular and virion-enclosed hepatitis C virus RNA are associated with peripheral-blood mononuclear cells in vivo. AB - BACKGROUND: Hepatitis C virus (HCV) replicating in peripheral-blood mononuclear cells (PBMCs) may represent an extrahepatic viral reservoir. Quantitation of HCV RNA with regard to its subcellular distribution and longitudinal course is needed for better understanding of the largely unexplored in vivo dynamics and potential pathogenetic significance of HCV in PBMCs. METHODS: Plasma and PBMCs from 30 patients coinfected with HCV and human immunodeficiency virus were evaluated in cross-sectional and longitudinal analyses, for up to 40 months. Differential extraction of virion-enclosed HCV RNA associated with cells was performed in parallel with extraction of total cellular HCV RNA. HCV RNA of either orientation was quantified by real-time polymerase chain reaction. RESULTS: HCV RNA was detected only in PBMCs from patients with viremia and at relatively stable quantities over time. Intracellular HCV RNA corresponding to ~60% of total cellular HCV RNA was strongly correlated with virion-enclosed HCV RNA but was only weakly associated with viral loads in plasma. In contrast, the ratio of HCV RNA load in PBMCs versus that in plasma was patient specific and stable over time. CONCLUSIONS: The substantial and patient-specific amounts of intracellular HCV RNA found by the present study support a concept of low-level replication in PBMCs. There was no evidence for persistent HCV infection in PBMCs after clearance of viremia in plasma. PMID- 17109345 TI - A complete mutational fitness map of the hepatitis C virus nonstructural 3 protease: relation to recognition by cytotoxic T lymphocytes. AB - The hepatitis C virus nonstructural (NS) 3/4A protease sequence is highly conserved for reasons not fully understood. We determined the protease activity in 181 NS3/4A gene products in which each protease residue was replaced by alanine or glycine. Unexpectedly, most (87%) protease residues could be replaced and protease activity would be retained. Using these data, we were able to identify a human leukocyte antigen A2-restricted epitope in which substitutions at 5 of 9 residues destroyed the protease. The NS3 protease shows an unexpectedly high plasticity, and it is therefore important to identify target sequences in which the appearance of mutations is restricted by viral fitness. PMID- 17109346 TI - Reductions in cross-neutralizing antibody responses in infants after attenuation of the human rotavirus vaccine candidate 89-12. AB - The G1P1A[8] rotavirus vaccine candidate 89-12, the precursor to Rotarix, stimulated high titers of neutralizing antibodies to non-G1/P1A[8] serotypes of human rotavirus in naturally infected subjects before attenuation by cell-culture passages. These responses were greatly diminished in young infants (median age, 11 weeks) administered the attenuated vaccine. Because of the possibility of improved responses in older infants, the immunogenicity of the 89-12 vaccine candidate was evaluated after administration of 2 doses beginning at either 4 or 6 months of age. As was found in young infants, neutralizing antibody responses to non-G1/P1A[8] rotaviruses were considerably lower than those observed after natural infection. The reasons identified were overall (P<.0001) lower neutralizing antibody responses stimulated by the attenuated 89-12 strain, compared with those stimulated by its virulent precursor, and 5 mutations selected in the gene encoding the immunodominant VP4 (P) neutralization protein. Even so, the Rotarix vaccine developed from attenuated 89-12 was found to elicit excellent protection against non-G1 rotaviruses. PMID- 17109347 TI - Quantification of viral inactivation by photochemical treatment with amotosalen and UV A light, using a novel polymerase chain reaction inhibition method with preamplification. AB - BACKGROUND: In evaluating a photochemical treatment process for inactivating parvovirus B19, there lacked simple culture methods to measure infectivity. The recently developed enzyme-linked immunospot (ELISpot) infectivity assay uses late stage erythropoietic progenitor cells and is labor intensive and time consuming. We evaluated a novel, efficient polymerase chain reaction (PCR) inhibition assay and examined correlations with reductions in infectivity. METHODS: Contaminated plasma was treated with 150 micromol/L amotosalen and 3 J/cm(2) ultraviolet A light and then tested for DNA modification using conventional PCR inhibition and a novel preamplification approach. The novel assay subjected the samples to preamplification cycles using long-template PCR, followed by quantitative PCR (QPCR) inhibition detection. Both approaches were tested for correlations with reductions in viral infectivity by comparing ELISpot assay results of identical samples. RESULTS: The B19 preamplification inhibition assay showed detection ranges of 2-2.5 log and demonstrated quantitative correlation with up to a 5.8 log reduction in viral infectivity in ELISpot results. Conventional PCR detected a >5 log reduction in amplification, correlated with a 4.4-log reduction in viral infectivity. A range of 4-log inhibition of hepatitis B virus DNA amplification was also achieved. CONCLUSIONS: The results demonstrated that a novel preamplification QPCR assay is a useful tool for predicting reductions in infectivity after photochemical treatment. This assay was extended to show utility in circumstances where practical in vitro assays are unavailable for the determination of the efficacy of pathogen inactivation. PMID- 17109348 TI - Clinical and immunologic risk factors for meningococcal C conjugate vaccine failure in the United Kingdom. AB - BACKGROUND: The meningococcal serogroup C conjugate (MCC) vaccine was introduced into the United Kingdom with licensure based on immunogenicity data not efficacy data. METHODS: All subjects with laboratory-confirmed meningococcal serogroup C (MenC) disease from January 2000 to December 2003 in England and Wales were followed up. A vaccine failure was defined as a laboratory-confirmed case of MenC disease occurring > or =10 days after the subject's last scheduled dose of MCC vaccine. Total immunoglobulins, serum bactericidal antibody (SBA) titers, MCC anticapsular antibody levels, and avidity indices (AIs) were measured in acute and convalescent serum samples from subjects with vaccine failure and unvaccinated subjects with MenC disease. RESULTS: Of 465 subjects with confirmed MenC disease identified among those eligible for vaccination, information on vaccination history was obtained for 462 (99.4%); of these, 53 were subjects with vaccine failure. SBA titers in convalescent serum samples and AIs in acute serum samples were significantly higher in subjects with vaccine failure than in unvaccinated subjects, (6.1-fold higher for SBA titers [P=.03] and 3.2-fold higher for AIs [P=.001]). CONCLUSIONS: The antibody response in the subjects with vaccine failure was consistent with an anamnestic response, suggesting that MenC disease occurred despite the MCC vaccine priming for immune memory. Persistence of antibodies may be a more appropriate correlate of long-term protection for MCC vaccines than the ability to generate a booster response on exposure. PMID- 17109349 TI - Human cytolytic T cell recognition of Yersinia pestis virulence proteins that target innate immune responses. AB - Cell contact by the plague bacterium Yersinia pestis initiates the injection of several virulence factors that target biochemical pathways critical for host clearance of bacteria. Despite this impairment of innate immunity, it is unclear whether antigen recognition by T cells is equally affected. We present evidence that human cytolytic T cells respond to Y. pestis virulence proteins presented by infected monocytes and dendritic cells. These T cell antigens consisted of a panel of proteins encoded by pCD1, a 70-kDa plasmid that harbors virulence factors and transport proteins of the cell contact-dependent, type III secretion system. Infected cells retained the ability to process and present tetanus toxoid to T cells, which indicates that responses to unrelated antigens were also maintained. Our results indicate that T cell immunity remains functional during Y. pestis infection, which thus suggests the potential benefits of therapeutic vaccination and strategies that emphasize the inclusion of cytotoxic T lymphocyte responses. PMID- 17109350 TI - Is Panton-Valentine leukocidin the major virulence determinant in community associated methicillin-resistant Staphylococcus aureus disease? AB - Methicillin-resistant Staphylococcus aureus (MRSA) remains a major problem in hospitals, and it is now spreading in the community. A single toxin, Panton Valentine leukocidin (PVL), has been linked by epidemiological studies to community-associated MRSA (CA-MRSA) disease. However, the role that PVL plays in the pathogenesis of CA-MRSA has not been tested directly. To that end, we used mouse infection models to compare the virulence of PVL-positive with that of PVL negative CA-MRSA representing the leading disease-causing strains. Unexpectedly, strains lacking PVL were as virulent in mouse sepsis and abscess models as those containing the leukotoxin. Isogenic PVL-negative (lukS/F-PV knockout) strains of USA300 and USA400 were as lethal as wild-type strains in a sepsis model, and they caused comparable skin disease. Moreover, lysis of human neutrophils and pathogen survival after phagocytosis were similar between wild-type and mutant strains. Although the toxin may be a highly linked epidemiological marker for CA-MRSA strains, we conclude that PVL is not the major virulence determinant of CA-MRSA. PMID- 17109351 TI - Antibiotic selection may contribute to increases in macrolide-resistant Treponema pallidum. AB - To determine whether the 23S rRNA mutation that confers macrolide resistance is present in >1 Treponema pallidum strain, 58 isolates collected between 2001 and 2005 were screened for this mutation and for an unrelated sequence that distinguishes between strains. The odds of identifying a macrolide-resistant strain increased over time (P=.006). In subjects who had received macrolides in the previous year, the relative risk of harboring a resistant strain was 2.2 (95% confidence interval, 1.1-4.4; P=.02). The macrolide-resistant strains were not identical. These findings suggest that macrolide resistance may be increasing in multiple strains in response to antibiotic pressure. PMID- 17109352 TI - Uropathogenic Escherichia coli alters muscle contractions in rat urinary bladder via a nitric oxide synthase-related signaling pathway. AB - BACKGROUND: Uropathogenic Escherichia coli (UPEC) is a common cause of urinary tract infection. The mechanisms by which bacteria cause the symptoms of cystitis remain unclear. METHODS: The contractions of isolated rat detrusor strips evoked by electrical field stimulations (EFS) or by exogenous agonists and immunoblotting for the detection of protein expression in the bladder were measured in the short (1 h) and long (24 h) term after the intravesical instillation of J96 (O4:K6) strain or UPEC isolated from patients with acute E. coli pyelonephritis or E. coli lipopolysaccharide (LPS). RESULTS: One hour after the instillation of UPEC, the level of endothelial nitric oxide synthase (eNOS) and the contractile response, but not protein kinase C (PKC) and extracellular signal-regulated kinase (ERK)-1/2 activation, were higher. Twenty-four hours after UPEC treatment, detrusor contractions were decreased, and inducible (i) NOS protein expression and ERK1/2 phosphorylation, but not PKC activation, were observed. Both aminoguanidine and PD98059 treatment markedly reversed the decrease of EFS- and acetylcholine-evoked detrusor contractions induced by UPEC. The instillation of LPS triggered PKC activation but not ERK1/2 phosphorylation. CONCLUSIONS: Short-term intravesical instillation of UPEC enhances detrusor contractions through an eNOS-related pathway, but iNOS-regulated ERK1/2 signaling may be involved in long-term UPEC treatment-induced responses. There are different mechanisms involved in the responses induced by UPEC and LPS. PMID- 17109353 TI - Evaluation of purified Taenia solium glycoproteins and recombinant antigens in the serologic detection of human and swine cysticercosis. AB - Cysticercosis caused by infection with embryonated eggs of Taenia solium is an important cause of neurological disease worldwide. On the basis of mitochondrial DNA analysis, T. solium is divided into 2 (African/American and Asian) genotypes. Glycoproteins (GPs) in cyst fluid purified from the 2 genotypes of T. solium were characterized and compared with the recombinant chimeric T. solium-Ag1V1/Ag2 protein (Rec-Ag1V1/Ag2) as serodiagnostic antigens. Immunoblot analysis revealed that banding patterns of GPs differed between the 2 genotypes because of posttranslation modification, especially glycosylation. The comparison of native GPs with Rec-Ag1V1/Ag2 by enzyme-linked immunosorbent assay demonstrated that there was no statistical difference in sensitivity. In addition, the conservation of the genes encoding Ag1V1 and Ag2 in T. solium worldwide was verified. These results indicate that Rec-Ag1V1/Ag2 has great potential for usefulness in serodiagnosis as an alternative to native antigens. PMID- 17109354 TI - Increase in serogroup C meningococcal disease in Canada is associated with antigenic changes in the protein antigens of the ET-15 clone of Neisseria meningitidis. PMID- 17109356 TI - Is there a safe age for vaccinating infants with tetravalent rhesus-human reassortant rotavirus vaccine? PMID- 17109358 TI - Is human herpesvirus-6 DNA in plasma the right marker for active infection? PMID- 17109360 TI - Recurrence of symptomatic bacterial vaginosis 12 months after oral metronidazole therapy in HIV-positive and -negative women. PMID- 17109362 TI - Natriuretic peptides levels are related to HDL-cholesterol with no influence on endothelium dependent vasodilatation. AB - BACKGROUND: The natriuretic peptides, Brain Natriuretic Peptide (BNP), C-type Natriuretic Peptide (CNP), are mediators of cardiovascular homeostasis. The impairment of arterial ability to vasodilate, also known as endothelial dysfunction, represents the first stage of atherosclerotic damage and may be assessed as brachial flow mediated vasodilation (FMV) in human. Generally an altered brachial FMV is documented in association to several cardiovascular risk factors as hypercholesterolemia. Aim of the study was to evaluate the behaviour of BNP and CNP in hyperlipemia and the potential relationship to FMV. PATIENTS AND METHODS: Forty-four hyperlipemic patients (LDL-cholesterol > 130 mg/dl and/or triglycerides > 150, age 35-60 y) of both genders and 20 normolipemic patients, matched for age and sex were investigated. RESULTS: Patients had lower values of brachial FMV in comparison to controls (3.9 +/- 3.5 vs 7.5 +/- 0.5%, p < 0.005), no differences were observed in BNP (4.6 +/- 4.6 vs 5.9 +/- 3.4 ng/mL, p = n.s) and CNP (4.1 +/- 5.8 vs 5.7 +/- 3.3 ng/mL, p = n.s). Univariate analysis showed a positive correlation between BNP and HDL-cholesterol values (r = 0.36, p = 0.001). In the multivariate analysis, LDL-cholesterol (beta = -0.57), HDL cholesterol (beta = 0.26) and brachial artery diameter (beta = -0.33) were predictors of brachial FMV. The only predictive variable for CNP was HDL cholesterol (beta = 0.37). CONCLUSIONS: The present study suggested that natriuretic peptides, BNP and CNP, are not altered in patients affected by hypercholesterolemia. Nevertheless, the levels of HDL-cholesterol are strictly related to the values of CNP. This observation, in humans, adds another mechanism to the vascular control exerted by HDL. PMID- 17109363 TI - Is C-reactive protein a better predictor of recurrent carotid disease following carotid endarterectomy than established risk factors for atherosclerosis? AB - BACKGROUND: The study was undertaken to identify possible relations between established and novel risk factors for atherosclerosis (gender, age, diabetes mellitus, hypertension, smoking and C-reactive protein) and the possibility of carotid restenosis following carotid endarterectomy (CEA). PATIENTS AND METHODS: A prospective study of 193 consecutive patients, admitted electively for carotid endarterectomy during 68 months, was conducted. 131 patients had symptomatic and 62 asymptomatic carotid disease. An attempt was made to follow-up on all operated arteries with duplex sonography at 2 months, 6 months, 12 months, and 24 months postoperatively. The correlation of previously mentioned variables (gender, age, diabetes mellitus, hypertension, smoking and average values of C-reactive protein) with progressive or recurrent disease was determined by chi-square analysis and analysis of variance. RESULTS: Of all 193 examined patients 29 demonstrated increasing degree of artery stenosis, while recurrent artery stenosis of > 69% was diagnosed in 11 patients. Age, gender, hypertension, and diabetes did not play a significant role in the presence of progressive (or recurrent) disease, while active smokers and patients with preoperative and average C-reactive protein (CRP) levels over 3.0 mg/L had a greater propensity to develop progression (or recurrence) of carotid disease. CONCLUSIONS: Increased levels of CRP in serum may be a better predictor of carotid restenosis after CEA than other established risk factors for vascular disease--except active smoking. PMID- 17109364 TI - Revisiting the frequency of peripheral arterial disease in patients with coronary artery disease: is there a difference between diabetic and non-diabetic patients? AB - BACKGROUND: The aim of this study was to investigate whether frequency of concomitant peripheral arterial disease (PAD) is associated with angiographic severity of coronary artery disease (CAD), as well as to ascertain if diabetic patients differ from those without diabetes in the association between these two manifestations of atherosclerosis. PATIENTS AND METHODS: This study included 302 patients (229 men, mean age 62.2 +/- 11.5 years) with documented CAD, divided into groups I-III, according to the angiographic severity of coronary atherosclerosis. Group I comprised 140 patients (104 men) with severe CAD, group II comprised 63 patients (48 men) with moderate CAD and group III comprised 99 patients (77 men) with mild CAD. Each of the groups I-III was further divided into the subgroups of diabetic and non-diabetic patients. Included were also 88 patients (42 men, mean age 61.7 +/- 9.5 years) without CAD and a control group of 60 healthy volunteers (30 men), aged 18-40 years. PAD was diagnosed by means of a Doppler apparatus. RESULTS: Frequency of PAD was associated with angiographic severity of CAD (p = 0.0001). This association was shown both in diabetic (p = 0.012) and in non-diabetic patients (p = 0.0041). Significantly (p < or = 0.01) higher frequency of PAD among diabetic patients was found in each of the groups I III. CONCLUSIONS: Among patients with CAD, frequency of concomitant PAD is associated with angiographic severity of coronary atherosclerosis. This association is demonstrated both in diabetic and in non-diabetic patients. Finally, PAD is significantly more frequent in diabetic patients, irrespective of the angiographic severity of CAD. PMID- 17109365 TI - General versus locoregional anesthesia in carotid surgery: a prospective randomised trial. AB - BACKGROUND: To evaluate the influence of anesthetic technique on perioperative neurological and cardiopulmonary complication rates in patients undergoing carotid endarterectomy. PATIENTS AND METHODS: 186 patients with symptomatic internal carotid artery (ICA) stenosis > 70% or asymptomatic ICA stenosis > 80% were prospectively randomized for either locoregional (LA) or general anesthesia (GA). RESULTS: Neurological complication rates were similar in both groups (GA 2% vs. LA 2%). Cardiopulmonary complication rates were not significantly different (GA 4% vs LA 1%). There were no stroke-related deaths, but one patient from the GA group died from severe postoperative pneumonia. Thus, a significant difference in combined stroke/cardiopulmonary related death between the two groups (GA 1% vs LA 0%) could not be found. However, perioperative cardiopulmonary monitoring showed that significantly more patients operated under general anesthesia had hypertensive events, with systolic blood pressure values greater than 180 mmHg on postoperative day one. There were no differences in the number of postoperatively hypotensive episodes (systolic blood pressure values < 100 mmHg) between the two groups. CONCLUSIONS: Significant differences in the perioperative neurological and cardiopulmonary complication rates between general and locoregional anesthesia in patients undergoing carotid endarterectomy could not be observed. PMID- 17109366 TI - [Simultaneous manifestation of blue-rubber-bleb-nevus-syndrome and malignant melanoma]. AB - Blue rubber bleb nevi (BRBN) are rare vascular malformations of the integument and mucous membranes. Little is known about coherence between these nevi and malignant diseases. We report on a young man with progressive BRBN syndrome representing a thitherto unknown malignant melanoma. PMID- 17109367 TI - Efficient haemodialysis despite complete central venous thrombosis. AB - Two cases with a fully functional haemodialysis access, in spite of complete thrombotic occlusion of the ipsilateral subclavian and/or brachiocephalic vein are reported. The coincidentally detected complete venous occlusions may indicate that occult venous stenosis or thrombosis is more frequent than generally assumed. In order to avoid deterioration of the haemodynamic situation interventions may be withheld unless clinical problems related to a diminished outflow occur. PMID- 17109368 TI - [Endovascular treatment of a thoraco-abdominal aortic aneurysm in case of a specific aortitis]. AB - Successful endovascular repair of an aortic aneurysm of the descending thoracic aorta, laying directly cranial of the celiac artery, caused by a spondylodiscitis of the thoracoabdominal spine. While vascular surgeons refused the open resection of the infected aneurysm, endovascular treatment with a stent-graft was performed. Respecting the celiac artery by catheterisation, endovascular treatment was managed without occluding the A. Adamkiewicz. In case of an unknown infection peri- and postoperative treatment was performed of a prolonged antibiotic and corticosteroid therapy. In a follow-up of three months period, there was a complete regression of the inflammatory aneurysm and an improvement of the spondylodiscitis. PMID- 17109369 TI - Abdominal aortic aneurysm in association with horseshoe kidney. AB - We report the preoperative diagnosis and surgical treatment of an abdominal aortic aneurysm (AAA) in association with a horseshoe kidney (HSK) in a 70-year old man. Through a median laparotomy a vascular tube graft was successfully used for repair the AAA. The extensive parenchymal isthmus overlying the aneurysm remained intact. PMID- 17109370 TI - [Acute limb ischemia as first symptom by contained ruptured pseudoaneurysm of an undifferentiated high grade pleomorphic sarcoma of the arteria iliaca externa]. AB - Primary malignant vascular tumors represent a rare cause of acute extremity ischemia. Due to unspecific symptoms the correct diagnosis is often delayed and confirmed in many cases as late as post mortem. Differential diagnosis of malignant vascular tumors should be considered in patients with acute ischemia, atypical history and absence of typical risk factors for vascular diseases. The overall prognosis of such tumors is poor. If possible, complete curative resection in combination with arterial reconstruction should be performed. Multimodal therapy has to be considered and discussed in appropriate tumor boards. We report a case of a 70-year-old male patient with acute ischemia and contained rupture of a pseudoaneurysm of the external iliac artery due to an undifferentiated high grade pleomorphic sarcoma. At the time of the primary operation, diffuse skeletal metastases were present but even detected postoperatively during staging. Therefore, no adjuvant or palliative therapy was initiated. In the postoperative course, recurrent non reversible ischemia was present followed by amputation of the right leg. The patient died 5 months after first operation. In the autopsy further metastases of lung and liver were found. PMID- 17109371 TI - Gastric predominant hereditary hemorrhagic telangiectasia. AB - Hereditary hemorrhagic telangiectasia (HHT) or Rendu-Osler-Weber disease is an autosomal-dominant disorder that frequently presents with epistaxis and gastrointestinal bleeding which may be a diagnostic and therapeutic challenge. We describe a 48-year-old monk with familiar history of HHT, who presented with frequent epistaxes and symptoms of ferropenic anemia. Upper gastrointestinal endoscopy revealed innumerable telangiectasias in the stomach. The patient underwent three sessions with argon plasma coagulation treatment but the results were poor. We also review the literature on the pathophysiology of the disease and discuss the suggested treatment. PMID- 17109375 TI - The statistical performance of an MCF-7 cell culture assay evaluated using generalized linear mixed models and a score test. AB - Biological assays often utilize experimental designs where observations are replicated at multiple levels, and where each level represents a separate component of the assay's overall variance. Statistical analysis of such data usually ignores these design effects, whereas more sophisticated methods would improve the statistical power of assays. This report evaluates the statistical performance of an in vitro MCF-7 cell proliferation assay (E-SCREEN) by identifying the optimal generalized linear mixed model (GLMM) that accurately represents the assay's experimental design and variance components. Our statistical assessment found that 17beta-oestradiol cell culture assay data were best modelled with a GLMM configured with a reciprocal link function, a gamma error distribution, and three sources of design variation: plate-to-plate; well to-well, and the interaction between plate-to-plate variation and dose. The gamma distributed random error of the assay was estimated to have a coefficient of variation (COV) = 3.2 per cent, and a variance component score test described by X. Lin found that each of the three variance components were statistically significant. The optimal GLMM also confirmed the estrogenicity of five weakly oestrogenic polychlorinated biphenyls (PCBs 17, 49, 66, 74, and 128). Based on information criteria, the optimal gamma GLMM consistently out-performed equivalent naive normal and log-normal linear models, both with and without random effects terms. Because the gamma GLMM was by far the best model on conceptual and empirical grounds, and requires only trivially more effort to use, we encourage its use and suggest that naive models be avoided when possible. PMID- 17109376 TI - Micromixer utilizing electrokinetic instability-induced shedding effect. AB - This paper presents a T-shaped micromixer featuring 45 degrees parallelogram barriers (PBs) within the mixing channel. The presented device obtains a rapid mixing of two sample fluids with conductivity ratio of 10:1 (sample concentration:running buffer concentration) by means of the electrokinetic instability-induced shedding effects which are produced when a direct current (DC) electric field of an appropriate intensity is applied. The presented device uses a single high-voltage power source to simultaneously drive and mix the sample fluids. The effectiveness of the mixer is characterized experimentally as a function of the applied electrical field intensity and the extent to which the PBs obstruct the mixing channel. The experimental results indicate that the mixing performance reaches 91% at a cross-section located 2.3 mm downstream of the T-junction when the barriers obstruct 4/5 of the channel width and an electrical field of 300 V/cm is applied. The micromixing method presented in this study provides a simple low-cost solution to mixing problems in lab-on-a-chip systems. PMID- 17109377 TI - Experimental and numerical investigation into leakage effect in injectors of microfluidic devices. AB - This paper performs an experimental and numerical investigation into low-leakage injectors designed for electrophoresis microchips. The principal material transport mechanisms of electrokinetic migration, fluid flow, and diffusion are considered in developing a mathematical model of the electrophoresis process. Low leakage injectors designed with injection channels orientated at various included angles are designed and tested. The numerical and experimental results indicate that the injector with a 30 degrees included angle successfully minimizes sample leakage and has an exciting potential for use in high-quality, high-throughput chemical analysis procedures and in many other applications in the field of micro total analysis systems. PMID- 17109378 TI - Atmospheric molding of ionic copolymer MALDI-TOF/MS arrays: a new tool for protein identification/profiling. AB - An atmospheric molding protocol has been used to prepare an ionic methacrylate based copolymer sample support chips for MALDI (pMALDI)-MS by targeting selected groups of various monomers copolymerized during molding, namely, carboxy, sulfo, dimethylalkyamino, and trimethylalkylammonium groups. The new disposable array chips provide analyte-oriented enhancement of protein adsorption to the modified substrates without requiring complicated surface coating or derivatization. The MALDI-MS performance of the new ionic copolymer chips was evaluated for lysozyme, beta-lactoglobulin A, trypsinogen and carbonic anhydrase I using washing with solutions prepared in pH or ionic strength steps. On cationic chips, the proteins are washed out at pH lower than their p/ values, and on anionic chips at pH higher than their p/ values. The ability of the microfabricated pMALDI chip set to selectively adsorb different proteins from real samples and to significantly increase their MS-signal was documented for the transmembrane photosystem I protein complex from the green alga Chlamydomonas reinhardtii. The proteins were almost exclusively adsorbed according to calculated pI values and grand average of hydropathy (GRAVY) indexes. The new disposable chips reduce manipulation times and increase measurement sensitivity for real-world proteomic samples. The simple atmospheric molding procedure enables additional proteomic operations to be incorporated on disposable MALDI-MS integrated platforms. PMID- 17109379 TI - Post-translational modification accounts for the presence of varied forms of nerve growth factor in Australian elapid snake venoms. AB - The Australian elapid snakes are amongst the most venomous snakes in the world, but much less is known about the overall venom composition in comparison to Asian and American snakes. We have used a combined approach of cDNA cloning and 2-DE with MS to identify nerve growth factor (NGF) in venoms of the Australian elapid snakes and demonstrate its neurite outgrowth activity. While a single 730 nucleotide ORF, coding for a 243 amino acid precursor protein was detected in all snakes, use of 2-DE identified NGF proteins with considerable variation in molecular size within and between the different snakes. The variation in size can be explained at least in part by N-linked glycosylation. It is possible that these modifications alter the stability, activity and other characteristics of the snake NGFs. Further characterisation is necessary to delineate the function of the individual NGF isoforms. PMID- 17109380 TI - Membrane microdomains and proteomics: lessons from tetraspanin microdomains and comparison with lipid rafts. AB - Biological membranes are compartmentalized into microdomains that exhibit particular lipid and protein compositions. Membrane microdomains, such as tetraspanin-enriched microdomains and lipid rafts, have been suggested to play a role in a variety of physiological and pathological processes. Therefore, the characterization of the protein compositions of these microdomains, which is the focus of this review, appears to be a crucial step to better understanding their function. Proteomics has recently allowed the characterization of tetraspanin enriched microdomains in colon cancer cells. This demonstrated the presence of different categories of membrane proteins and suggested a variation in the composition of tetraspanin-enriched microdomains during tumor progression. On the other hand, proteomics has permitted the identification of hundreds of proteins in lipid rafts of different origins. However, the diversity of methodologies in sample preparation and of strategies in protein identification led to a broad variability in the data obtained. These methodological issues are discussed. Moreover, proteomics has revealed that different sets of proteins were present in tetraspanin-enriched microdomains as compared with lipid rafts, strengthening the idea that these microdomains are distinct structures. PMID- 17109381 TI - Discrimination of intact mycobacteria at the strain level: a combined MALDI-TOF MS and biostatistical analysis. AB - New methodologies for surveillance and identification of Mycobacterium tuberculosis are required to stem the spread of disease worldwide. In addition, the ability to discriminate mycobacteria at the strain level may be important to contact or source case investigations. To this end, we are developing MALDI-TOF MS methods for the identification of M. tuberculosis in culture. In this report, we describe the application of MALDI-TOF MS, as well as statistical analysis including linear discriminant and random forest analysis, to 16 medically relevant strains from four species of mycobacteria, M. tuberculosis, M. avium, M. intracellulare, and M. kansasii. Although species discrimination can be accomplished on the basis of unique m/z values observed in the MS fingerprint spectrum, discrimination at the strain level is predicted on the relative abundance of shared m/z values among strains within a species. For the 16 mycobacterial strains investigated in the present study, it is possible to unambiguously identify strains within a species on the basis of MALDI-TOF MS data. The error rate for classification of individual strains using linear discriminant analysis was 0.053 using 37 m/z variables, whereas the error rate for classification of individual strains using random forest analysis was 0.023 using only 18 m/z variables. In addition, using random forest analysis of MALDI TOF MS data, it was possible to correctly classify bacterial strains as either M. tuberculosis or non-tuberculous with 100% accuracy. PMID- 17109382 TI - Escherichia coli proteome chips for detecting protein-protein interactions. PMID- 17109383 TI - Changes of the hepatic proteome in murine models for toxically induced fibrogenesis and sclerosing cholangitis. AB - We investigated the changes in the hepatic proteome in murine models for toxic induced fibrogenesis and sclerosing cholangitis. A comprehensive comparison of protein changes observed is made and the mechanistical basis of the expression changes is discussed. Hepatic fibrosis was induced by repetitive intraperitoneal CCl4 treatment of BALB/c mice or developed spontaneously in BALB/c-ATP-binding cassette, subfamily B, member 4 (Abcb4) knock out mice. Fibrosis was verified by a morphometric score and assessment of hydroxyproline content of liver tissue, respectively. The innovative difference in-gel electrophoresis (DIGE) technique was used to analyse protein expression levels of the mouse proteome. Results were confirmed by Western blotting and real-time RT-PCR. In CCl4-induced fibrosis 20 out of 40 and in BALB/c-Abcb4(-/-) mice 8 out of 28 differentially expressed proteins were identified utilizing DIGE. Only two proteins, selenium-binding protein (Sbp2) and carbonic anhydrase 3, have been unidirectionally expressed (i.e. down-regulated) in both models. Relevant differences in the pathogenesis of toxically induced liver fibrosis and sclerosing cholangitis exist. The only novel protein with regard to liver fibrosis depicting a unidirectional expression pattern in both animal models was Sbp2. An explicit protein function could not be clarified yet. PMID- 17109384 TI - Risk factors for intrahepatic cholangiocarcinoma in a low-risk population: a nationwide case-control study. AB - Recently, the incidence of intrahepatic cholangiocarcinoma (ICC) has been increasing in a number of developed (Western) countries. However, risk factors in these low-risk populations are poorly understood. In this nationwide population based case-control study in Denmark, we examined the relationship between selected medical conditions and subsequent ICC risk to provide additional clues to etiopathogenesis. All histologically confirmed ICC cases diagnosed in Denmark between 1978 and 1991 were identified from the Danish cancer registry. Population controls were selected from the central population registry and were matched 4:1 to cases on sex and year of birth. Cases and controls were linked to the Danish hospital discharge registry to obtain information on prior hospital diagnoses. Odds ratios (OR) and 95% confidence intervals (95% CI) were derived using conditional logistic regression. A total of 764 ICC cases and 3,056 population controls were included in the study. Chronic liver diseases were significantly related to ICC: alcoholic liver disease (OR = 19.22, 95% CI = 5.55-66.54), unspecified cirrhosis (OR = 75.9, 95% CI 10.2-565.7). Bile duct diseases were also associated with risk: cholangitis (OR = 6.3, 95% CI = 2.3-17.5), choledocholithiasis (OR = 23.97, 95% CI = 2.9-198.9), cholecystolithiasis (OR = 4.0, 95% CI = 2.0-7.99), though gallbladder removal did not change risk (OR = 1.6, 95% CI = 0.65-3.7). Among other conditions, chronic inflammatory bowel disease (OR = 4.7, 95% CI = 1.65-13.9) was significantly associated with ICC. Diabetes was associated with risk in the year prior to diagnosis of ICC (OR = 3.02, 95% CI = 1.05-8.69). Obesity was unrelated to risk. These results confirm that prior bile duct diseases increase risk of ICC and suggest that alcoholic liver disease and diabetes may also increase risk. PMID- 17109385 TI - Safety and tolerability of a novel chromatography-based intravenous immunoglobulin when administered at a high infusion rate in patients with immune thrombocytopenic purpura. AB - Intravenous immunoglobulin (IGIV) therapy is generally considered to be a safe and effective treatment for idiopathic thrombocytopenic purpura (ITP). The usual initial treatment dose is 1-2 g/kg body weight, which results in an extended infusion time, significantly impacting patients' day-to-day activities. Two crossover studies assessed the safety and tolerability of a novel IGIV preparation (IGIV-C; Gamunex, 10%) when infused at rates ranging from 0.08 mL/kg/min (the standard maximum licensed rate) to 0.14 mL/kg/min in patients with ITP. The first study included 28 patients and 3 infusion rates; 0.08, 0.11, and 0.14 mL/kg/min. The second study included 8 patients and 2 infusion rates; 0.08 and 0.14 mL/kg/min were evaluated. The incidence of infusion-related adverse events was similar for all infusion rates. Headache was the most commonly reported infusion-related adverse event. The incidence, combined for Studies 1 and 2, was 14.7% (n=34), 18.2% (n=22), and 19.4% (n=31) of patients, for each infusion rate of 0.08, 0.11, and 0.14 mL/kg/min, respectively. The majority were mild in severity. None of the other drug-related, treatment-emergent events were serious; most were mild, in spite of the higher rate of fluid loading over a shorter period of time for patients infused at 0.14 mL/kg/min. There were no clinically important changes in parameters that distinguished between infusion rates; there were no signs of hemolysis. The results suggest that IGIV-C infused at rates up to 0.14 mL/kg/min in patients with ITP is well tolerated. PMID- 17109386 TI - Lepirudin for anticoagulation in patients with heparin-induced thrombocytopenia treated with continuous renal replacement therapy. AB - Lepirudin is a potent, direct thrombin inhibitor used for anticoagulation in patients with heparin-induced thrombocytopenia type II (HIT). The half-life of lepirudin is prolonged in patients with renal insufficiency. Preliminary studies suggest that it is safe to use lepirudin in patients being treated with intermittent hemodialysis but information regarding its use with continuous renal replacement therapy (CRRT) is scarce. CRRT is used in acute care settings to remove fluid and uremic toxins in patients with renal failure with hemodynamic instability. Patients with HIT, renal failure, and hemodynamic instability pose a complex situation for clinical management. These patients require anticoagulation with nonheparin agents with simultaneous CRRT. There are no guidelines in the literature regarding the management of this patient group. We report our experience with lepirudin at managing four such patients with HIT, being treated with CRRT. PMID- 17109387 TI - Autosomal recessive von Willebrand disease associated with compound heterozygosity for a novel nonsense mutation (2908 del C) and the missense mutation C2362F: definite evidence for the non-penetrance of the C2362F mutation. AB - A novel null mutation (2908del C in exon 22) of the von Willebrand factor (VWF) gene was identified in compound heterozygosity with the missense mutation G7335T (C2362F) in exon 42 in a propositus from a new family with autosomal recessive von Willebrand disease (VWD). The propositus, referred at age 2 for severe epistaxis and prolonged bleeding after a tongue bite, had factor VIII:C 14-21 IU/dL, VWF Antigen 3-8 IU/dL and Ristocetin Cofactor activity < 3 IU/dL. Multimeric pattern showed the lack of triplet pattern and a faster mobility of central band, while heterozygotes for C2362F showed intermediate mobility compared to normal plasma and plasma from the propositus. In the propositus' family 5 subjects were heterozygotes for the C2362F mutation and 5 were heterozygotes for the cytosine deletion. Bleeding score was assessed with a detailed questionnaire in 28 heterozygotes for C2362F, 23 of whom identified in 5 previously reported families and 5 in the present one, and found to be similar to what is observed in normal controls and heterozygotes for null allele. In conclusion, the mutation C2362F is frequently observed in compound heterozygosity with null alleles in patients with recessive VWD in the Veneto region and cause bleeding only in the compound heterozygous or homozygous state. PMID- 17109389 TI - Early relapse of JAK2 V617F-positive chronic neutrophilic leukemia with central nervous system infiltration after unrelated bone marrow transplantation. AB - Chronic neutrophilic leukemia (CNL) is a rare myeloproliferative disorder characterized by a proliferation mainly of mature neutrophils. The prognosis is generally poor and an optimal therapeutic strategy remains to be determined. Allogeneic hematopoietic stem cell transplantation (HSCT) is expected to be the only curative therapy so far. We report a 46-year-old male with progressive CNL who underwent bone marrow transplantation from an HLA-matched unrelated donor. After engraftment was achieved on day 35, relapse of CNL was confirmed on day 50. The progression of CNL was very rapid afterward and infiltration to the central nervous system was observed. The Janus Kinase 2 (JAK2) V617F homozygous mutation was detected from the peripheral blood or bone marrow samples throughout the clinical course. From comparison with reports of successful HSCT for CNL in the literature, it was inferred that HSCT should be performed in a stable status before progression. Furthermore, JAK2 V617F-positive CNL may contain an aggressive disease entity in contrast to previous reports. Accumulation of experiences is required to establish a definite role of HSCT in the treatment of CNL and a prognostic significance of JAK2 mutation in CNL. PMID- 17109390 TI - The effect of budesonide mouthwash on oral chronic graft versus host disease. AB - Oral chronic graft versus host disease (cGVHD) is common and a major cause of morbidity and loss of quality of life in long term survivors. Cyclosporine with prednisone remains the first line therapy for oral manifestations of cGVHD. However, even with routine administration of systemic agents, many patients with oral manifestations of cGVHD do not have resolution of their disease and may benefit from incorporation of local therapy. Budesonide is a highly potent steroid which has minimal systemic side effects and being used for oral cGVHD. We designed a retrospective study to compare treatment results of patients with oral cGVHD who received topical budesonide in addition to systemic therapy that consists of combined prednisone and cyclosporine (Group A, n = 12), with the treatment results of patients who were administered the same systemic therapy alone (Group B, n = 11) to determine whether budesonide mouthwash had any advantage on response rates. Three mg topical budesonide/10 ml saline was used 3 4 times a day for up to 6 months in group A. Diagnosis, clinical staging, and treatment response scoring for cGVHD were performed according to National Institutes of Health (NIH) consensus criteria. At the baseline examination, there were no statistically significant differences in terms of median oral cGVHD examination scores between two groups. After treatment, there was statistically significant decrease in median oral cGVHD examination scores compared to baseline (P < 0.001 and 0.021), and significant differences were found between two groups (P < 0.032). Overall response rate was 83% and 36% for group A and B, respectively (P = 0.036). However, no statistically significant differences were found between median pain scores of two groups before and after treatment (P = 0.740 and P = 0.091). No major systemic side effects and oral candidiasis were observed in two groups of patients. We concluded that topical budesonide might be added to systemic therapy to obtain better response rates in patients with oral cGHVD. PMID- 17109391 TI - Allelic loss analysis by denaturing high-performance liquid chromatography and electrospray ionization mass spectrometry. AB - Analysis of allelic imbalance is of great importance for understanding tumorigenesis and the clinical management of malignant disease. Fluorescent-based capillary electrophoresis (CE) of highly polymorphic short tandem repeats (STRs) has become the main method used to detect the loss/gain of alleles. However, there is continued interest in the development of techniques that require no fluorescence and allow the rapid analysis of individual samples. One promising alternative is ion-pair reversed-phase high-performance liquid chromatography (IP RP-HPLC), which is widely available because of its use in denaturing HPLC. Its applicability in combination with ultraviolet (UV) absorbance detection to the efficient separation of di- and tetranucleotide repeats on the short arm of chromosome 11 was tested using 25 matched pairs of normal and ovarian cancer tissues. Loss of heterozygosity (LOH) could be readily identified for all 13 loci tested, based on changes in the ratios between either the alleles or homo- and heteroduplex signals. However, discrimination between noninformative homo- or hemizygous and heterozygous samples was difficult or impossible when HPLC failed to resolve the alleles. Hyphenation of HPLC with electrospray ionization (ESI) quadrupole ion trap (IT) mass spectrometry (MS) not only allowed the identification of coeluting alleles, but also the reliable detection of a 40% reduction of one allele. The size range of DNA fragments amenable to mass spectrometric analysis was effectively tripled to >300 bp by the use of a linear IT and a Taq DNA polymerase cocktail lacking detergents that otherwise adversely affect ESI. PMID- 17109392 TI - Exploring the pyridoxal 5'-phosphate-dependent enzymes. AB - Pyridoxal 5'-phosphate (PLP)-dependent enzymes represent about 4% of the enzymes classified by the Enzyme Commission. The versatility of PLP in carrying out a large variety of reactions exploiting the electron sink effect of the pyridine ring, the conformational changes accompanying the chemical steps and stabilizing distinct catalytic intermediates, and the spectral properties of the different coenzyme-substrate derivatives signaling the reaction progress, are some of the features that have attracted our interest to investigate the structure-dynamics function relationships of PLP-dependent enzymes. To this goal, an integrated approach combining biochemical, biophysical, computational, and molecular biology methods was used. The extensive work carried out on two enzymes, tryptophan synthase and O-acetylserine sulfhydrylase, is presented and discussed as representative of other PLP-dependent enzymes we have investigated. Finally, perspectives of PLP-dependent enzymes functional genomics and drug targeting highlight the continuous novelty of an "old" class of enzymes. PMID- 17109393 TI - Life without water: expression of plant LEA genes by an anhydrobiotic arthropod. AB - Anhydrobiotic animals protect cellular architecture and metabolic machinery in the dry state, yet the molecular repertoire supporting this profound dehydration tolerance is not fully understood. For the desiccation-tolerant crustacean, Artemia franciscana, we report differential expression of two distinct mRNAs encoding for proteins that share sequence similarities and structural features with late-embryogenesis abundant (LEA) proteins originally discovered in plants. Bioinformatic analyses support assignment of the LEA proteins from A. franciscana to group 3. This eucoelomate species is the most highly evolved animal for which LEA gene expression has been reported. It is becoming clear that an ensemble of micromolecules and macromolecules is important for establishing the physical conditions required for cellular stabilization during drying in nature. PMID- 17109394 TI - The search for non-chordate retinoic acid signaling: lessons from chordates. AB - Signaling by retinoic acid (RA) is an important pathway in the development and homeostasis of vertebrate and invertebrate chordates, with a critical role in mesoderm patterning. Classical studies on the distribution of nuclear receptors of animals suggested that the family of RA receptors (RARs/NR1B) was restricted to chordates, while the family of RA X receptors (RXR/NR2B) was distributed from cnidarians to chordates. However, the accumulation of data from genome projects and studies in non-model species is questioning this traditional view. Here we discuss the evidence for non-chordate RA signaling systems in the light of recent advances in our understanding of carotene (pro-Vitamin A) metabolism and of the identification of potential RARs and members of the NR1 family in echinoderms and lophotrochozoan trematodes, respectively. We conclude, as have others before (Bertrand et al., 2004. Mol Biol Evol 21(10):1923-1937), that signaling by RA is more likely an ancestral feature of bilaterians than a chordate innovation. PMID- 17109395 TI - Population-based birth defects surveillance systems in the United States. PMID- 17109396 TI - Birth defects surveillance data from selected states, 1999-2003. PMID- 17109398 TI - Targeted gene delivery to differentiated skeletal muscle: a tool to study dedifferentiation. AB - Cellular dedifferentiation is required for functional regeneration in salamanders. Dedifferentiating multinucleate skeletal muscle gives rise to mononucleate cells during limb regeneration. Efficient methods and tools must be developed in order to understand the molecular cues underlying dedifferentiation. Here we describe a non-viral method to express extra-chromosomal DNA exclusively in terminally differentiated muscle without the need for cell purification steps. After cytoplasmic injection of various expression vectors into myotubes or myofibres, we detect long-lasting mRNA and protein expression in up to 70% of the injected cells. The combination of the transfection protocol with live imaging allows a time- and cost-effective screen of candidate genes in terminally differentiated muscle cells of both amphibian and mammalian origin. PMID- 17109399 TI - State birth defects surveillance program directory. PMID- 17109400 TI - Spatial arrangement of metal nanoparticles supported by porous polymer substrates studied by transmission electron microtomography. AB - Fine metal particles (nanoparticles) stabilized on porous (polymeric) substrates can be considered as a model system of a high-performance catalyst. In the present study, the substrate was made using the periodic microphase-separated structure of a block copolymer as the template, and the Pd nanoparticles were formed inside the porous material by reduction of the Pd2+ ions with 1-propanol as the reductant. The three-dimensional morphology of such a polymer-Pd hybrid material was studied by transmission electron microtomography. The characteristic structural parameters of the hybrid, e.g., the penetration of the Pd nanoparticles into the polymer substrate, number density of the Pd nanoparticles, and size distribution of the Pd nanoparticles, were measured for the first time. PMID- 17109401 TI - Sonohysterography is superior to transvaginal sonography for the diagnostic approach of irregular uterine bleeding in women of reproductive age. AB - PURPOSE: To evaluate and compare the accuracy of transvaginal sonography (TVS) and sonohysterography (SHG) in the investigation of women of reproductive age presenting with irregular uterine bleeding (IUB). METHODS: This prospective study included 104 women presenting with IUB. All patients underwent TVS, SHG, and hysteroscopy, during which endometrial biopsies were obtained and any endometrial mass was treated with hysteroscopic surgery. Statistical analysis was performed by calculating the sensitivity, specificity, and positive and negative predictive values of TVS and SHG in diagnosing endometrial polyp, submucous myoma and all endometrial pathologies (polyp, submucous myoma, endometrial hyperplasia, and endometrial carcinoma) with the histopathological report of the tissues obtained by hysteroscopy serving as the end point for the analysis. RESULTS: The sensitivity, specificity, and positive and negative predictive values, respectively of TVS were 61.2%, 90.9%, 85.7%, and 72.5% for diagnosing endometrial polyps; 75.0%, 92.0%, 63.1%, and 95.3% for diagnosing submucous myomas; and 75.0%, 80.6%, 87.9%, and 63.0% for diagnosing any kind of pathology. The corresponding diagnostic values of SHG were 83.7%, 96.4%, 95.3%, and 86.9% for polyps; 87.5%, 98.9%, 93.3%, and 97.8% for submucous myomas; and 88.2%, 91.7%, 95.2%, and 80.5% for any kind of pathology. CONCLUSIONS: SHG showed superior sensitivity, specificity, and positive and negative predictive values compared with TVS in diagnosing intrauterine lesions in women of reproductive age with IUB. PMID- 17109402 TI - Use of transrectal ultrasound-guided biopsy in the diagnosis of pelvic malignancies. AB - PURPOSE: To describe our experience with transrectal ultrasound (TRUS)-guided needle biopsy of pelvic malignancies. METHODS: Eleven patients with clinical suspecion of advanced malignant pelvic tumor were referred to our institution with a history of unsuccessful CT-guided biopsy, although a target lesion was demonstrated on pelvic CT or MRI. Cholin-PET and FDG-18-PET were also obtained individually in each patient. TRUS was performed using a commercially available three-dimensional scanner. Biopsies were performed with an 18G biopsy gun. In 9 of 11 patients, biopsy was successfully performed under analgesia, whereas general anesthesia was required in the other 2 patients. RESULTS: The lesions were identified with TRUS in all patients, and biopsies were taken successfully under TRUS guidance. In all patients, the harvested material was of excellent quality and was adequate for definitive pathological diagnosis. Pathological results included 6 nodal metastases from transitional cell carcinoma, 1 case of lymph node metastasis from prostate cancer, 1 paravesical recurrence of cervical cancer, 1 metastasis from cecal cancer, and 2 cases of paravesical metastasis of a gastric cancer. CONCLUSION: TRUS-guided biopsy is a useful technique for the diagnosis of pelvic malignancies. It is faster and less expensive than CT-guided biopsy, and in most cases sufficient material can be harvested for a definitive pathological diagnosis. PMID- 17109403 TI - Theoretical model of human apolipoprotein B100 tertiary structure. AB - Low density lipoprotein (LDL) particles are the main cholesterol carriers in human plasma. The organization of the particle, composed of apolar lipids and phospholipid monolayer stabilized by apolipoprotein B100 (apoB), is highly complex and still unknown. ApoB is an extremely large protein (4563 amino acids) and very little is known about its structure. A 3D model of the N-terminal region has been recently proposed and has provided interesting insights about the physico-chemical properties of the protein and putative interaction zones with lipids. In the present article, we propose the first tentative 3D modelling for most remaining residues. All predicted features emerging from the models are confronted with agreement to experimental data available. Using different up-to date prediction methods, we decomposed the protein into eight domains and predicted 3D structure for each of them. The analysis of hydrophobic patches, polar regions, coupled with functional predictions based on the 3D models, gives new clues to understanding of the functional role of apoB. We suggest precise regions putatively involved in the lipid interactions, and discuss the position of apoB on the LDL particle. Finally, we propose relative organization of the domains, providing a shape quite compatible with the low resolution electron microscopy map. PMID- 17109404 TI - Automatic procedure for using models of proteins in molecular replacement. AB - In a crystallography experiment, a crystal is irradiated with X-rays whose diffracted waves are collected and measured. The reconstruction of the structure of the molecule in the crystal requires knowledge of the phase of the diffracted waves, information that is lost in the passage from the three-dimensional structure of the molecule to its diffraction pattern. It can be recovered using experimental methods such as heavy-atom isomorphous replacement and anomalous scattering or by molecular replacement, which relies on the availability of an atomic model of the target structure. This can be the structure of the target protein itself, if a previous structure determination is available, or a computational model or, in some cases, the structure of a homologous protein. It is not straightforward to predict beforehand whether or not a computational model will work in a molecular replacement experiment, although some rules of thumb exist. The consensus is that even minor differences in the quality of the model, which are rather difficult to estimate a priori, can have a significant effect on the outcome of the procedure. We describe here a method for quickly assessing whether a protein structure can be solved by molecular replacement. The procedure consists in submitting the sequence of the target protein to a selected list of freely available structure prediction servers, cluster the resulting models, select the representative structures of each cluster and use them as search models in an automatic phasing procedure. We tested the procedure using the structure factors of newly released proteins of known structure downloaded from the Protein Data Bank as soon as they were made available. Using our automatic procedure we were able to obtain an interpretable electron density map in more than half the cases. PMID- 17109405 TI - Learning about protein hydrogen bonding by minimizing contrastive divergence. AB - Defining the strength and geometry of hydrogen bonds in protein structures has been a challenging task since early days of structural biology. In this article, we apply a novel statistical machine learning technique, known as contrastive divergence, to efficiently estimate both the hydrogen bond strength and the geometric characteristics of strong interpeptide backbone hydrogen bonds, from a dataset of structures representing a variety of different protein folds. Despite the simplifying assumptions of the interatomic energy terms used, we determine the strength of these hydrogen bonds to be between 1.1 and 1.5 kcal/mol, in good agreement with earlier experimental estimates. The geometry of these strong backbone hydrogen bonds features an almost linear arrangement of all four atoms involved in hydrogen bond formation. We estimate that about a quarter of all hydrogen bond donors and acceptors participate in these strong interpeptide hydrogen bonds. PMID- 17109406 TI - Description of atomic burials in compact globular proteins by Fermi-Dirac probability distributions. AB - We perform a statistical analysis of atomic distributions as a function of the distance R from the molecular geometrical center in a nonredundant set of compact globular proteins. The number of atoms increases quadratically for small R, indicating a constant average density inside the core, reaches a maximum at a size-dependent distance R(max), and falls rapidly for larger R. The empirical curves turn out to be consistent with the volume increase of spherical concentric solid shells and a Fermi-Dirac distribution in which the distance R plays the role of an effective atomic energy epsilon(R) = R. The effective chemical potential mu governing the distribution increases with the number of residues, reflecting the size of the protein globule, while the temperature parameter beta decreases. Interestingly, betamu is not as strongly dependent on protein size and appears to be tuned to maintain approximately half of the atoms in the high density interior and the other half in the exterior region of rapidly decreasing density. A normalized size-independent distribution was obtained for the atomic probability as a function of the reduced distance, r = R/R(g), where R(g) is the radius of gyration. The global normalized Fermi distribution, F(r), can be reasonably decomposed in Fermi-like subdistributions for different atomic types tau, F(tau)(r), with Sigma(tau)F(tau)(r) = F(r), which depend on two additional parameters mu(tau) and h(tau). The chemical potential mu(tau) affects a scaling prefactor and depends on the overall frequency of the corresponding atomic type, while the maximum position of the subdistribution is determined by h(tau), which appears in a type-dependent atomic effective energy, epsilon(tau)(r) = h(tau)r, and is strongly correlated to available hydrophobicity scales. Better adjustments are obtained when the effective energy is not assumed to be necessarily linear, or epsilon(tau)*(r) = h(tau)*r(alpha,), in which case a correlation with hydrophobicity scales is found for the product alpha(tau)h(tau)*. These results indicate that compact globular proteins are consistent with a thermodynamic system governed by hydrophobic-like energy functions, with reduced distances from the geometrical center, reflecting atomic burials, and provide a conceptual framework for the eventual prediction from sequence of a few parameters from which whole atomic probability distributions and potentials of mean force can be reconstructed. PMID- 17109407 TI - MUPRED: a tool for bridging the gap between template based methods and sequence profile based methods for protein secondary structure prediction. AB - Predicting secondary structures from a protein sequence is an important step for characterizing the structural properties of a protein. Existing methods for protein secondary structure prediction can be broadly classified into template based or sequence profile based methods. We propose a novel framework that bridges the gap between the two fundamentally different approaches. Our framework integrates the information from the fuzzy k-nearest neighbor algorithm and position-specific scoring matrices using a neural network. It combines the strengths of the two methods and has a better potential to use the information in both the sequence and structure databases than existing methods. We implemented the framework into a software system MUPRED. MUPRED has achieved three-state prediction accuracy (Q3) ranging from 79.2 to 80.14%, depending on which benchmark dataset is used. A higher Q3 can be achieved if a query protein has a significant sequence identity (>25%) to a template in PDB. MUPRED also estimates the prediction accuracy at the individual residue level more quantitatively than existing methods. The MUPRED web server and executables are freely available at http://digbio.missouri.edu/mupred. PMID- 17109409 TI - Plasma deposition of tetraglyme inside small diameter tubing: optimization and characterization. AB - In this study, a glow discharge plasma deposition system previously used for treating flat substrates was successfully modified and optimized to produce a PEO like coating on the inner surface of 1-3 mm ID polyethylene tubing by deposition of tetra ethylene glycol dimethyl ether (tetraglyme). The plasma treatment conditions were varied in order to find operating values that would produce coatings with the ultralow (< 5 ng/cm(2)) fibrinogen adsorption (Gamma(Fg)) previously shown necessary to significantly reduce platelet adhesion. The flow rate of gaseous tetraglyme monomer, pressure, and plasma generating power were found to be the most important parameters affecting the uniformity and chemical structure of the coating. The coating uniformity and quality were assessed by measuring Gamma(Fg) at positions 1 cm apart along the entire tube and the fraction of C1s carbon that was in an ether bond (ether-carbon ratio) by electron spectroscopy of chemical analysis. Under optimized conditions, tetraglyme plasma coated tubes of up to 20 cm in length had ultralow Gamma(Fg). The region of the tube that had ultralow Gamma(Fg) also had C1s ether-carbon ratios that are greater than 50%. PMID- 17109408 TI - How a small change in retinal leads to G-protein activation: initial events suggested by molecular dynamics calculations. AB - Rhodopsin is the prototypical G-protein coupled receptor, coupling light activation with high efficiency to signaling molecules. The dark-state X-ray structures of the protein provide a starting point for consideration of the relaxation from initial light activation to conformational changes that may lead to signaling. In this study we create an energetically unstable retinal in the light activated state and then use molecular dynamics simulations to examine the types of compensation, relaxation, and conformational changes that occur following the cis-trans light activation. The results suggest that changes occur throughout the protein, with changes in the orientation of Helices 5 and 6, a closer interaction between Ala 169 on Helix 4 and retinal, and a shift in the Schiff base counterion that also reflects changes in sidechain interactions with the retinal. Taken together, the simulation is suggestive of the types of changes that lead from local conformational change to light-activated signaling in this prototypical system. PMID- 17109410 TI - Three-dimensional growth behavior of osteoblasts on biomimetic hydroxylapatite scaffolds. AB - The authors used rapid prototyping to produce three-dimensional hydroxylapatite scaffolds with controlled, fully interconnected porosity. The purpose of this study was to illuminate the effect of hormones on the osteogenic differentiation and to investigate how osteoblasts colonize the three-dimensional scaffold focusing on the formation of the cellular network. Preosteoblasts were seeded onto scaffolds, were optionally treated with the osteogenic hormones triiodo-L thyronine (T3) and 1,25-dihydroxyvitamin-D3 (D3), and the expression of osteoblastic marker genes was investigated. Confocal laser scanning microscopy was used to investigate the three-dimensional growth behavior. Culturing cells on scaffolds strongly increased the expression of osteocalcin, osteoprotegerin, Runx2, and receptor activator of NFkB-ligand (RANKL). Treatment with T3 increased the expression of osteocalcin but did not change that of osteoprotegerin and Runx2. Treatment with D3 inhibited the expression of osteocalcin, Runx2, and osteoprotegerin. Both hormones had similar effects in the three-dimensional system as found in two-dimensional cultures although more accentuated, indicating that preosteoblasts behave more naturally on three-dimensional structures. The osteoblasts colonized the three-dimensional squared pores of scaffolds by forming a cellular network with a round central channel keeping it into the depth and depositing collagen fibrils. These results provide insight how osteoblasts colonize a three-dimensional system and underline the importance of this environment in osteoblastic differentiation studies. PMID- 17109411 TI - In vivo response to starch-based scaffolds designed for bone tissue engineering applications. AB - Our purpose was to evaluate the in vivo endosseous response to three starch-based scaffolds implanted in rats (n = 54). We implanted the three scaffold groups; a 50/50 (wt %) blend of corn starch and ethylene-vinyl alcohol (SEVA-C), the same composition coated with a biomimetic calcium phosphate (Ca-P) layer (SEVA-C/CaP), and a 50/50 (wt %) blend of corn starch and cellulose acetate (SCA), all produced by extrusion with blowing agents, into distal femurs proximal to the epiphyseal plate, for 1, 3, or 6 weeks. Our results showed that at 1 week considerable reparative bone formed around all scaffold groups, although the bone was separated from the scaffold by an intervening soft tissue interfacial zone that comprised two distinct compartments: the surface of the scaffold was occupied by multinucleate giant cells and the compartment between these cells and the surrounding bone was occupied by a streaming fibrous-like tissue. The extracellular matrix of the latter was continuous with the extracellular bone matrix itself, labeled positively for osteocalcin and appeared mineralized by back-scattered electron imaging. All three scaffolds showed a similar tissue response, with the soft tissue interface diminishing with time. No bone contact was observed with SEVA-C at any time point, only transitory bone contact was observed with SEVA-C/CaP at 3 weeks, but SCA exhibited direct bone contact at 6 weeks where 56.23 +/- 6.46% of the scaffold surface was occupied by bone. We conclude that all materials exhibited a favorable bony response and that the rapidly forming initial "connective tissue" seen around all scaffolds was a very early form of bone formation. PMID- 17109412 TI - A novel ordered nano hydroxyapatite coating electrochemically deposited on titanium substrate. AB - A novel porous nano hydroxyapatite (HA) coating has been prepared on commercially pure titanium substrate by a modified electrochemical deposition method. The physico-chemical and biological properties of the coating were characterized by SEM, XRD, FTIR, Raman, and in vitro cell culture test respectively. The SEM patterns show a uniform microporous morphology consisting of wirelike crystals at nanometer scale. It is suggested that under controlled deposition conditions, the primary HA nanowires grow and self-assemble to construct an ordered microporous nest-like morphology, thus to form a nano-micro two-level structure. The XRD results demonstrate that the HA nanowires are orderly arranged with their c-axis preferentially perpendicular to the substrate surface. The Raman and IR spectra affirm that the main component of the coating is well crystallized HA. An interdigitation phenomenon of the MG63 human osteosarcoma cells with the HA nanowires is observed in the in vitro test, indicating excellent biocompatibility and bioactivity for the prepared coating. PMID- 17109413 TI - Depth-dependent mechanical properties of enamel by nanoindentation. AB - Nanoindentation has recently emerged to be the primary method to study the mechanical behavior and reliability of human enamel. Its hardness and elastic modulus were generally reported as average values with standard deviations that were calculated from the results of multiple nanoindentation testing. In such an approach, it is assumed that the mechanical properties of human enamel are constant, independent of testing parameters, like indent depth and loading rate. However, little is known if they affect the measurements. In this study, we investigated the dependence of the hardness and elastic modulus of human enamel on the indent depth. We found that in a depth range from 100 to 2000 nm the elastic moduli continuously decreased from approximately 104 to 70 GPa, and the hardnesses decreased from approximately 5.7 to 3.6 GPa. We then considered human enamel as a fiber-reinforced composite, and used the celebrated rule of mixture theory to quantify the upper and lower bounds of the elastic moduli, which were shown to cover the values measured in the current study and previous studies. Accordingly, we attributed the depth dependence of the hardness and modulus to the continuous microstructure evolution induced by the nanoindenter tip. PMID- 17109414 TI - Investigation of in vitro biocompatibility of novel pentablock copolymers for gene delivery. AB - Novel pentablock copolymers of poly(diethylaminoethylmethacrylate) (PDEAEM), poly(ethylene oxide) (PEO), and poly(propylene oxide) (PPO), (PDEAEM-b-PEO-b-PPO b-PEO-b-PDEAEM), were synthesized as vectors for gene delivery, and were tested for their biocompatibility on SKOV3 (human ovarian carcinoma) and A431 (human epidermoid cancer) cell lines under different in vitro conditions using various assays to elucidate the mechanism of cell death. These copolymers form micelles in aqueous solutions and can be tuned for their cytotoxicity by tailoring the weight percentage of their cationic component, PDEAEM. Copolymers with higher PDEAEM content were found to be more cytotoxic, though their polyplexes were less toxic than the polycations alone. Pentablock copolymers displayed higher cell viability than commercially available ExGen 500 at similar N:P ratios. While cell death with ExGen was found to be accompanied by an early loss of cell membrane integrity, pentablock copolymers caused very little membrane leakage. Caspase-3/7 assay confirmed that none of these polymers induced apoptosis in the cells. These pentablock copolymers form thermo-reversible gels at physiological temperatures, thereby enabling controlled gene delivery. Toxicity of the polymer gels was tested using an agarose-matrix, simulating an in vivo tumor model where injected polyplex gels would dissolve to release polyplexes, diffusing through tumor mass to reach the target cells. Twenty five weight percent of copolymer gels were found to be nontoxic or mildly cytotoxic after 24 h incubation. Transfection efficiency of the copolymers was found to be critically correlated to cytotoxicity and depended on DNA dose, polymer concentration, and N:P ratios. Transgene expression obtained was comparable to that of ExGen, but ExGen exhibited greater cell death. PMID- 17109415 TI - Preclinical biocompatibility assessment of the EVAHEART ventricular assist device: coating comparison and platelet activation. AB - Thromboembolism and bleeding remain significant complications of ventricular assist device (VAD) support. Increasing the amount of biocompatibility data collected during preclinical studies can provide additional criteria to evaluate device refinements, while design changes may be implemented before entering clinical use. Twenty bovines were implanted with the EVAHEART centrifugal VAD for durations from 30 to 196 days. Titanium alloy pumps were coated with either diamond-like carbon or 2-methoxyethyloylphosphoryl choline (MPC). Activated platelets and platelet microaggregates were quantified by flow cytometry, including two new assays to quantify bovine platelets expressing CD62P and CD63. Temporally, all assays were low preoperatively, then significantly increased following VAD implantation, before declining to a lower, but still elevated level over 2-3 weeks. MPC-coated VADs produced significantly fewer activated platelets after implant trauma effects diminished. Three animals receiving no postoperative anticoagulation had similar amounts of circulating activated platelets and platelet microaggregates as animals receiving warfarin anticoagulation. Two new methods to quantify bovine activated platelets using antibodies to CD62P and CD63 were characterized and applied. These measures, along with previously described assays, were able to differentiate between two biocompatible coatings and assess effects of anticoagulation regimen in VAD preclinical testing. PMID- 17109416 TI - Influence of silanated filler content on the biodegradation of bisGMA/TEGDMA dental composite resins. AB - It has been shown that an increase in the content of nonsilanated submicron colloidal silica filler particles within dental composites resulted in the release of more bis-phenol-A diglycidyl dimethacrylate (bisGMA)-derived product, bis-hydroxy-propoxyphenyl propane, following incubation with cholesterol esterase (CE). This work further investigates the enzyme-catalyzed biodegradation of fine composite resin systems, containing silanated micron-size irregular glass fillers, commonly used in clinical restorations. Model composite resin samples (10 or 60% weight fraction silanated barium glass filler, 1 mum average particle size) based on bisGMA/triethylene glycol dimethacrylate (TEGDMA) were incubated in buffer or buffer with CE (pH = 7.0, 37 degrees C) solutions for 32 days. The incubation solutions were analyzed using high-performance liquid chromatography, UV spectroscopy, and mass spectrometry. Both groups were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, and X-ray photoelectron spectroscopy. In contrast with previous findings for nonsilanated submicron filler systems, the higher filler containing composite showed an increase in its stability with time, following exposure to esterase and when compared to the lower filler content material. As well, the 60% filler composite leached less unreacted monomer TEGDMA. Since the model composite resins studied here were identical and only the filler content varied, the differences in biostability could be specifically associated with the relative amount of resin/filler distribution. The clinical use of different materials in varied dental applications (ranging from fissure sealant to tooth-colored highly filled materials) must consider the potential for different degradation profiles to occur as a function of filler content. PMID- 17109417 TI - Preparation and assessment of glutaraldehyde-crosslinked collagen-chitosan hydrogels for adipose tissue engineering. AB - A portfolio of crosslinked chitosan:collagen blends was prepared, and their microarchitecture and water binding capacity were studied to investigate their application for adipose tissue engineering. Glutaraldehyde (GA) concentration had little effect on scaffold morphology or water binding capacity. However, the processing freezing temperature prior to lyophilization affected both. In vitro cytocompatibility of pre-adipocytes (PAs) was assessed for a candidate collagen:chitosan blend using two assays. Results confirm the viability of PAs on GA-crosslinked collagen:chitosan scaffolds. A rat subcutaneous pocket assay was employed to assess PA-seeded scaffolds in vivo. Animal tests proved that PA seeded scaffolds were biocompatible, could induce vascularization, and form adipose tissue. PMID- 17109419 TI - Should we continue teaching anatomy by dissection when ...? AB - The central role that human dissection has long held in clinical education is being reevaluated in many institutions. Despite the impression that many institutions are abandoning dissection, very few have and most of those have reinstated dissection within a few years. What are the inherent qualities that lead institutions back to dissection? In our efforts to redesign a shortened dissection course, our consultations with a broad range of clinicians lead us to understand how the rhythms of clinical practice are modeled and developed in the small-group setting of the dissection laboratory. Following further consultation with colleagues who have experimented with different models of anatomy instruction, we discuss three themes in support of dissection. First, problem solving in the dissection laboratory develops the habits-of-mind of clinical practice. Second, relating dissection to imaging modalities develops the spatial reasoning skills needed to understand computer simulations, interpret imaging data, and interact with surgeons, radiologists, and patients. Third, the human face of dissection fosters self-reflection and integration of the cognitive and affective skills required for medical practice. Through group process, the collaborative effort of dissection teams develops essential of attributes of clinical professionalism. PMID- 17109420 TI - Race and the odd history of human paleontology. AB - Although the late 17th century witnessed the recognition of fossils as the remains of extinct organisms-because they could be incorporated into the creation story embodied in the Great Chain of Being-acceptance of human antiquity through the indisputable demonstration of the contemporaneity of human bones, stone tools, and accepted fossils was not forthcoming for nearly 2 centuries thereafter. When it did occur, however, ancient humans were not seen as presenting a pattern of diversity similar to that seen in the fossil records of nonhuman organisms. Instead, human evolution then, as now, has typically been interpreted as being unilinear. This belief can be traced to Huxley (1863), who argued that the Feldhofer Grotto Neanderthal skullcap was merely an extension into the past of morphology seen in the Australian Aborigine, whom he took to represent the primitive end of an extreme range of variation he thought characterized Homo sapiens. During the mid-20th century, Mayr and Dobzhansky (mis)used their clout as founders of the evolutionary synthesis to cement in paleoanthropology the idea that human evolutionary history was characterized by nonspeciation. As such, anything that could be interpreted as potentially representing taxic diversity was relegated to the status of individual variation. Lack of understanding of the history of human paleontology, and the biases that constrained its perspective on human evolution, continue to affect the ways in which most paleoanthropologists pigeonhole human fossils. PMID- 17109421 TI - Anatomy of reflux: a growing health problem affecting structures of the head and neck. AB - Gastroesophageal reflux disease (GERD) and laryngopharyngeal reflux (LPR) are sibling diseases that are a modern-day plague. Millions of Americans suffer from their sequelae, ranging from subtle annoyances to life-threatening illnesses such as asthma, sleep apnea, and cancer. Indeed, the recognized prevalence of GERD alone has increased threefold throughout the 1990s. Knowledge of the precise etiologies for GERD and LPR is becoming essential for proper treatment. This review focuses on the anatomical, physiological, neurobiological, and cellular aspects of these diseases. By definition, gastroesophageal reflux (GER) is the passage of gastric contents into the esophagus; when excessive and damaging to the esophageal mucosa, GERD results. Reflux that advances to the laryngopharynx and, subsequently, to other regions of the head and neck such as the larynx, oral cavity, nasopharynx, nasal cavity, paranasal sinuses, and even middle ear results in LPR. While GERD has long been identified as a source of esophageal disease, LPR has only recently been implicated in causing head and neck problems. Recent research has identified four anatomical/physiological "barriers" that serve as guardians to prevent the cranial incursion of reflux: the gastroesophageal junction, esophageal motor function and acid clearance, the upper esophageal sphincter, and pharyngeal and laryngeal mucosal resistance. Sequential failure of all four barriers is necessary to produce LPR. While it has become apparent that GER must precede both GERD and LPR, the head and neck distribution of the latter clearly separates these diseases as distinct entities warranting specialized focus and treatment. PMID- 17109424 TI - Reflections on eponyms in neuroscience terminology. AB - Eponyms have played a very significant linguistic role in technical and scientific terminology. They are an important feature of language that have contributed for a long time to engraving in history the names of those researchers who have devoted their lives to scientific discovery. In the field of medical terminology, they are an asset, although their semantic effectiveness has constituted a long-standing debate. We will analyze how language contributes to the advance of science and technology and the current position of eponyms in the health sciences. Eponymy in neuroscience has been used for a long time as a way to identify and recognize scientific issues, such as diseases, syndromes, methods, processes, substances, organs, and parts of organs as a way to honor those who, in a certain way, contributed to the progress of science. However, sometimes those honors do not correspond to the real contributors, thus receiving a nondeserved acknowledgment. Another problem with eponymic references is the lack of information about the matter in hand, because eponyms do not provide any clear information leading to the identification of the situation under study, as they are not reasonably descriptive. The aim of this article is to encourage the use of descriptive terms instead of eponyms and to establish a system of scientific nomenclature to consolidate the use of the language as a means of conveying scientific information among experts. PMID- 17109425 TI - Anatomy meets architecture: designing new laboratories for new anatomists. AB - General notions of architecture are familiar to anatomists, and they frequently use the word in describing the functional structures of cells, tissues, and whole organisms. Beyond concepts relating to orderly structure, anatomists infrequently encounter the profession of architecture and practicing architects. Significantly, anatomists can work with architects in the design and building of laboratories and classrooms, efforts that can have sustained effects on the practice of anatomy. In this paper, we consider cooperative interactions between anatomists and architects in designing new laboratories that accommodate educational innovations and increasingly valuable dissection resources. We begin by introducing architecture and architects in their roles in design and building. We next consider essential features and technologies for new laboratories that support a combination of classical dissection, prosection, models, and computer based information. Different working conditions are reviewed for designing renovations of existing facilities, long-term planning for new, same-institution buildings, and extramural planning and construction for new medical schools. Whatever the project, anatomists work with architects in repeated interactive planning meetings that arrive at working laboratory designs by a process similar to successive approximation. In consulting on designs for extramural institutions, anatomists must balance client administration and faculty needs with objective oversight of practice-side design features, constraints, and capacity for innovative uses with new curricula. Architects are the key agents in producing laboratories designed for flexible and innovative anatomical education, although client-favored models for Internet-based technology can limit future use of cadavers in multiyear teaching of medical and health sciences students. PMID- 17109426 TI - Anatomy of learning: instructional design principles for the anatomical sciences. AB - Teaching anatomy is becoming increasingly challenging due to the progressive evolution of university teaching missions, student populations, medical and undergraduate curricula, coupled with a paucity of empirically tested evidence based instructional practices in the anatomical and medical education literature. As a mechanism to confront these pedagogical challenges, recent advances in educational psychology are analyzed for developing a framework to guide educational reform efforts. Extensive research in educational psychology over the last 100 years has resulted in four major theories on human learning that have facilitated a paradigm shift from teacher-centered to learner-centered classrooms and are described here in temporal order of development: behavioral theory, information processing theory, metacognitive theory, and social constructivist theory. Each theory is analyzed in detail and is used to construct instructional design principles for enhancing anatomical education research and practice. An example of a cognitively based learning environment for an undergraduate anatomy course is presented. Preliminary results suggest that intentionally drawing on different theories of learning when making instructional decisions gave students the learning support they needed to be successful and nearly doubled the course's student retention rate over a 3-year period. PMID- 17109427 TI - Vertical bone augmentation with granulated brushite cement set in glycolic acid. AB - Brushite cements are a biocompatible materials that are resorbed in vivo. A new cement composed of a mixture of monocalcium phosphate (MCP) and beta-tricalcium phosphate (beta-TCP) that sets using glycolic acid (GA) was synthesized and characterized. After setting, the cement composition, derived from X-ray diffraction, was 83 wt % brushite and 17 wt % beta-TCP with an average brushite crystal size of about 2.6 +/- 1.4 microm. The cement has a diametral tensile strength of 2.9 +/- 0.7 MPa. Granules prepared from the set-cement were used as grafting material in bone defects on rabbit calvaria for evaluating in vivo its bone regeneration capacity. Considerable cement resorption, improvement in the bone mineral density, and bone neoformation was observed after 4 weeks of the granules' implantation. PMID- 17109428 TI - Comparison of the clinical and preclinical biocompatibility testing of dental materials: are the ISO usage tests meaningful? AB - International Organization for Standardization (ISO 10993 and 7405) guidelines recommends the preclinical screening of dental materials using non-human primates. The literature contains no comparisons of responses to dental materials. To test the accuracy of preclinical screening tests for predicting human clinical responses, 106 class V pulp exposed cavities were prepared in human and non-human primate teeth. Teeth were restored with calcium hydroxide and amalgam, zinc oxide eugenol or resin-modified glass ionomer. Teeth were extracted after 10-163 days and prepared for histological analysis. Pulp cell numbers were compared and their reactionary dentin activity measured in response to cavity preparation. Pulp inflammatory activity was categorized according to ISO standards. There were no statistically significant differences between human and non-human primate teeth in terms of pulp reactions to dental materials. The use of non-human primates for preclinical biocompatibility investigation provided an accurate method of evaluating clinical responses to dental materials. PMID- 17109429 TI - Direct encapsulation of BSA and DNA into silica microcapsules (hollow spheres). AB - We reported before that silica hollow spheres (microcapsules) are prepared by interfacial reaction method that W/O emulsion with the aqueous solution of sodium silicate and n-hexane solution of Tween 80 and Span 80 is combined with another aqueous solution of silica precipitant such as NH(4)HCO(3) and NH(4)Cl. This procedure using W/O/W emulsion fabricates the hollow structures of silica particles directly, and additional steps such as the removal of core parts, that are often essential for the preparation of hollow particles via core-shell materials, are not required. When biomacromolecules such as protein and nucleic acid are mixed in the aqueous solution of sodium silicate, these macromolecules can be encapsulated into the microcapsules. We succeeded the direct encapsulation of bovine serum albumin (BSA) and duplex DNA. Most of encapsulated BSA and DNA cannot be released from the microcapsules without the destruction of microcapsule shell. These microcapsule materials encapsulating biomacromolecules will be applied to biotechnologies such as immobilized enzyme and so on. PMID- 17109430 TI - The origin of the two-electron/four-centers C--C bond in pi-TCNE(2)2- dimers: electrostatic or dispersion? AB - The structure and stability of the pi-TCNE(2)2- dimers in K2TCNE2 aggregates is revisited trying to find if the origin of their two-electron/four-centers C--C bond are the electrostatic K+-TCNE- interactions or the dispersion interactions between the anions. The study is done at the HF, B3LYP, CASSCF (2,2), and MCQDPT/CASSCF (2,2) levels using the 6-31+G(d) basis set. Our results show that the only minima of this aggregate that preserves the pi-TCNE(2)2- structure has the two K+ atoms placed in equatorial positions in between the two TCNE- planes. When the K+ atoms are placed along the D2h axis of the anions the structure is not a minimum. The main energetic component responsible for the stability of these aggregates comes from the cation-anion interactions. However, a proper accounting of the dispersion component (as done in the MCQDPT/CASSCF (2,2) calculations) is needed to make the closed-shell singlet more stable than the open-shell singlet. Thus, the bond results from the combination of the electrostatic and dispersion components, being the first the dominant one. The optimum geometry of the closed-shell singlet is very similar to the experimental one found in crystals. PMID- 17109431 TI - Atom-atom partitioning of total (super)molecular energy: the hidden terms of classical force fields. AB - Classical force fields describe the interaction between atoms that are bonded or nonbonded via simple potential energy expressions. Their parameters are often determined by fitting to ab initio energies and electrostatic potentials. A direct quantum chemical guide to constructing a force field would be the atom atom partitioning of the energy of molecules and van der Waals complexes relevant to the force field. The authors used the theory of quantum chemical topology to partition the energy of five systems [H2, CO, H2O, (H2O)2, and (HF)2] in terms of kinetic, Coulomb, and exchange intra-atomic and interatomic contributions. The authors monitored the variation of these contributions with changing bond length or angle. Current force fields focus only on interatomic interaction energies and assume that these purely potential energy terms are the only ones that govern structure and dynamics in atomistic simulations. Here the authors highlight the importance of self-energy terms (kinetic and intra-atomic Coulomb and exchange). PMID- 17109432 TI - Sigma and pi contributions to the induced magnetic field: indicators for the mobility of electrons in molecules. AB - The authors discuss the role of the sigma and pi contributions to the induced magnetic field for simple hydrocarbons containing a double or a triple bond, as well as for benzene and cyclobutadiene. While the magnetic field induced by the sigma electrons is short-ranged, the pi system is responsible for the formation of long-range cones. These cones influence the chemical shift of atoms by additional shielding (for aromatic) or deshielding (for antiaromatic molecules) contributions. While the hydrogen atoms of benzene are found to lie within the deshielded region of the magnetic field induced by the pi electrons, they are shielded by the total induced magnetic field. The induced magnetic field of the pi electrons support Pople's model on the basis of first-principles calculations. PMID- 17109433 TI - Pair interaction energy decomposition analysis. AB - The energy decomposition analysis (EDA) by Kitaura and Morokuma was redeveloped in the framework of the fragment molecular orbital method (FMO). The proposed pair interaction energy decomposition analysis (PIEDA) can treat large molecular clusters and the systems in which fragments are connected by covalent bonds, such as proteins. The interaction energy in PIEDA is divided into the same contributions as in EDA: the electrostatic, exchange-repulsion, and charge transfer energies, to which the correlation (dispersion) term was added. The careful comparison to the ab initio EDA interaction energies for water clusters with 2-16 molecules revealed that PIEDA has the error of at most 1.2 kcal/mol (or about 1%). The analysis was applied to (H2O)1024, the alpha helix, beta turn, and beta strand of polyalanine (ALA)10, as well as to the synthetic protein (PDB code 1L2Y) with 20 residues. The comparative aspects of the polypeptide isomer stability are discussed in detail. PMID- 17109434 TI - Unicorns in the world of chemical bonding models. AB - The appearance and the significance of heuristically developed bonding models are compared with the phenomenon of unicorns in mythical saga. It is argued that classical bonding models played an essential role for the development of the chemical science providing the language which is spoken in the territory of chemistry. The advent and the further development of quantum chemistry demands some restrictions and boundary conditions for classical chemical bonding models, which will continue to be integral parts of chemistry. PMID- 17109435 TI - The existence of secondary orbital interactions. AB - B3LYP/6-311+G** (and MP2/6-311+G**) computations, performed for a series of Diels Alder (DA) reactions, confirm that the endo transition states (TS) and the related Cope-TSs are favored energetically over the respective exo-TSs. Likewise, the computed magnetic properties (nucleus-independent chemical shifts and magnetic susceptibililties) of the endo- (as well as the Cope) TS's reveal their greater electron delocalization and greater aromaticity than the exo-TS's. However, Woodward and Hoffmann's original example is an exception: their endo-TS model, involving the DA reaction of a syn- with an anti-butadiene (BD), actually is disfavored energetically over the corresponding exo-TS; magnetic criteria also do not indicate the existence of SOI delocalization in either case. Instead, a strong energetic preference for endo-TSs due to SOI is found when both BDs are in the syn conformations. This is in accord with Alder and Stein's rule of "maximum accumulation of double bonds:" both the dienophile and the diene should have syn conformations. Plots along the IRC's show that the magnetic properties typically are most strongly exalted close to the energetic TS. Because of SOI, all the points along the endo reaction coordinates are more diatropic than along the corresponding exo pathways. We find weak SOI effects to be operative in the endo TSs involved in the cycloadditions of cyclic alkenes, cyclopropene, aziridine, cyclobutene, and cyclopentene, with cyclopentadiene. While the endo-TSs are only slightly lower in energy than the respective exo-TSs, the magnetic properties of the endo-TS's are significantly exalted over those for the exo-TS's and the Natural Bond Orbitals indicate small stabilizing interactions between the methylene cycloalkene hydrogen orbitals (and lone pairs in case of aziridine) with pi-character and the diene pi MOs. PMID- 17109436 TI - Bond electron pair: its relevance and analysis from the quantum chemistry point of view. AB - This paper first comments on the surprisingly poor status that Quantum Chemistry has offered to the fantastic intuition of Lewis concerning the distribution of the electrons in the molecule. Then, it advocates in favor of a hierarchical description of the molecular wave-function, distinguishing the physics taking place in the valence space (in the bond and between the bonds), and the dynamical correlation effects. It is argued that the clearest pictures of the valence electronic population combine two localized views, namely the bond (and lone pair) Molecular Orbitals and the Valence Bond decomposition of the wave-function, preferably in the orthogonal version directly accessible from the complete active space self consistent field method. Such a reading of the wave function enables one to understand the work of the nondynamical correlation as an enhancement of the weight of the low-energy VB components, i.e. as a better compromise between the electronic delocalization and the energetic preferences of the atoms. It is suggested that regarding the bond building, the leading dynamical correlation effect may be the dynamical polarization phenomenon. It is shown that most correlation effects do not destroy the bond electron pairs and remain compatible with Lewis' vision. A certain number of free epistemological considerations have been introduced in the development of the argument. PMID- 17109437 TI - Gilbert N. Lewis and the chemical bond: the electron pair and the octet rule from 1916 to the present day. AB - We describe the development of Lewis's ideas concerning the chemical bond and in particular the concept of the electron pair bond and the octet rule. We show that the concept of the electron pair bond has endured to the present day and is now understood to be a consequence of the Pauli principle. In contrast the octet rule is now regarded as much less important than was originally generally believed, although Lewis himself knew several exceptions and regarded it as less important than what he called the rule of two (the electron pair). The octet rule was more strongly promoted by Langmuir who is also responsible for the term covalent bond. However, many more exceptions to the octet rules than were known to Lewis are now known and the terms hypervalent and hypovalent used to describe such molecules are no longer particularly useful. Today it is realized that bonding electron pairs in many molecules are not as well localized as Lewis believed, nevertheless resonance structures, i.e., plausible alternative Lewis structures, are still often used to describe such molecules. Moreover electrons are not always found in pairs, as for example in linear molecules, which can, however, be satisfactorily described by Linnett's double quartet theory. The electron density distribution in a molecule can now be analyzed using the ELF and other functions of the electron density to show where electron pairs are most probably to be found in a molecule. PMID- 17109438 TI - Variations of the nutrient artery of the fibula. AB - In this study the authors aimed to show variations of the nutrient artery of the fibula, which is important for more effective vascularized fibular grafts. Thirty intact legs were examined to determine the number, location, and diameter of the nutrient arteries in the shaft. In 27 of these specimens (90%) there was single nutrient artery; two legs (6.6%) had double nutrient artery. In one leg (3.3%) no nutrient artery was observed. The number of nutrient arteries found on each of the surfaces was summarized as follows. Most of the nutrient arteries pierced the fibula at its medial crest, while only one entered from the posterior surface. Most nutrient arteries were near the middle third of the fibula. The most frequent diameter of the nutrient artery was 0.9-1.5 mm. The authors think that their results about the variations of the nutrient artery supplying the fibula will contribute to the clinical and vascular anatomy knowledge. PMID- 17109439 TI - Course variability of the phrenic nerve in the presence of an azygos lobe: two case reports. PMID- 17109440 TI - Traditional and virtual microscopy compared experimentally in a classroom setting. AB - The technology known as virtual microscopy is now widely available to medical students. A number of medical school histology and pathology course directors, including those at the Eastern Virginia Medical School, are exploring the question of how best to make use of this new tool. The current study compared the efficacy of teaching and testing one unit of histology, bone and cartilage, using two technologies, namely, virtual microscopy and traditional microscopy. Additionally, the study examined whether low, moderate, high moderate, or high achieving students perform more effectively in any of the learning/testing formats. In a completely randomized block design, 96 first semester, first-year medical students were grouped by previous exam quartiles then subsequently randomly assigned to one of four groups. Using identical laboratory exercises with highly comparable slides for each experimental group, half of the subjects learned the exercise with traditional microscopy and half learned with virtual microscopy. Subjects were further randomly subdivided into virtual or actual testing groups. The authors found no significant differences in test scores when they examined effects by learning group or by testing group, nor were there significant interaction effects. Student performance evaluated by previous exam quartile was significant (P < 0.001). That is, students who had scored in a particular quartile on a previous test tended to score in the same quartile on the bone and cartilage test regardless of learning or testing method. In a short opinion survey, students were polled to evaluate their experience. Student preferences for both learning and testing method varied widely. PMID- 17109441 TI - Origins of the gonadal artery: embryologic implications. AB - The gonadal arteries are paired vessels that usually originate from the abdominal aorta at the level of second lumbar vertebra. In 5-20% of cases, the gonadal artery has a high origin (superior to L2) and in 5-6% of cases it originates from the main or accessory renal artery. The latter is referred to here as an aberrant gonadal artery. Ninety-eight kidneys of 50 healthy potential renal transplant donors were prospectively studied by conventional angiography. The renal artery, either main or accessory, was detected and individually injected to highlight their perihilar divisions and possible extrarenal branches. The gonadal arteries were recorded if they originated from the renal arteries. We found that 39% (n = 38) of kidneys had at least one accessory renal artery. In 14 sides (14% of kidneys), the gonadal artery (11 right and 3 left) originated from the renal artery, either main (n = 5) or accessory (n = 9). Ten out of 14 kidneys with an aberrant gonadal artery had an associated accessory renal artery. In nine cases, the gonadal artery originated from the accessory renal artery, and in one case, although it originated from the main renal artery, the same kidney had an accessory arterial supply. The results of this study demonstrate that aberrant gonadal arteries tend to originate from kidneys that possess an accessory arterial supply. We hypothesize that aberrancies of the gonadal artery are a part of a common embryologic error resulting in the persistence of the future accessory renal arteries. We believe that this study is the first to hypothesize and study such an association with these arterial anomalies of the renal pedicle. PMID- 17109442 TI - St. George's University: an anatomist's experiences during Grenada's revolution and intervention. PMID- 17109443 TI - Association between clinical characteristics and risk-reduction interventions in women who underwent BRCA1 and BRCA2 testing: a single-institution study. AB - BACKGROUND: Women who are at increased risk for breast and ovarian cancers, especially BRCA1 and BRCA2 mutation carriers, face a myriad of risk-reduction options, including increased surveillance, chemoprevention, prophylactic oophorectomy, and prophylactic mastectomy. However, little is known about which clinical, demographic, or cancer-related factors are associated with risk reduction interventions. METHODS: The authors conducted a retrospective review of records for 554 women who had undergone testing at The University of Texas M. D. Anderson Cancer Center between 2000 and 2006 for deleterious BRCA1 and BRCA2 gene mutations. Data were collected on the risk-reduction interventions these women adopted after they underwent genetic testing. These data were tested for associations with demographic and clinical characteristics. RESULTS: Among the 554 women who underwent genetic testing for BRCA mutation, 78 were found to have a deleterious mutation in the BRCA1 gene, and 54 had a mutation in the BRCA 2 gene. Of the 554 women, 85 underwent prophylactic mastectomy, 30 prophylactic oophorectomy, and 52 both surgeries; 387 women opted for surveillance. Women who had BRCA mutations, a history of breast cancer or ductal carcinoma in situ (DCIS), or previous breast biopsies were more likely to have prophylactic surgery. Women with a family history of ovarian cancer were more likely to undergo prophylactic oophorectomy. Women with a personal history of ovarian cancer or advanced breast cancer were more likely to undergo surveillance only. Women with breast cancer who had had a total mastectomy as part of their prior breast cancer treatment underwent prophylactic mastectomy more frequently than women who either had breast-conserving surgery or no history of breast cancer. In multivariate analysis, only positive BRCA mutation carrier status was associated with having had prophylactic surgery. In addition, breast cancer history was significantly associated with prophylactic mastectomy. CONCLUSIONS: Women who were BRCA carriers, women who had a history of breast cancer, DCIS, or breast biopsy, or had a family history of ovarian cancer were more likely to have undergone surgery for cancer risk reduction. Women with ovarian cancer or advanced breast cancer were more likely to have undergone surveillance. PMID- 17109444 TI - A recombinant human RNASET2 glycoprotein with antitumorigenic and antiangiogenic characteristics: expression, purification, and characterization. AB - BACKGROUND: Human RNASET2 is a T2-RNase glycoprotein encoded by the RNASET2 gene, which is located on chromosome 6 (6q27). Deletion in 6q27 is associated with several human malignancies. METHODS: A synthetic RNASET2 gene that was optimized for expression in the yeast Pichia pastoris was designed according to the cDNA sequence and was cloned under the control of the methanol-induced promoter fused to the alpha-mating secretion peptide. The recombinant protein was purified from the culture supernatant of transformed P. pastoris through an affinity Sepharose concanavalin A column. Actin-binding activity was examined by membrane blotting using monoclonal mouse antiactin immunoglobulin M and by cross-linking in solution to G-actin using 1-[3-(dimethylamino)propyl]-3-ethyl-carboimide methiodide. The antiangiogenic activity of RNASET2 (from 0.5 microM to 10 microM) was assessed by a human umbilical vein endothelial (HUVE) cell assay in the presence of 1 microg/mL angiogenin, basic fibroblast growth factor (bFGF), or recombinant human vascular endothelial growth factor (VEGF). Cell colony formation was examined in human colon HT29 cancer cells to assess the antitumorigenic activity of RNASET2 or the enzymatic-inactivated RNASET2 (EI RNASET2) (1 microM each). In an athymic mouse xenograft model, LS174T human cancer cells were injected subcutaneously. When tumors were palpable, the mice were treated for 3 weeks with RNASET2 (1 mg/kg), paclitaxel (10 mg/kg or 15 mg/kg), or a combination of the 2 drugs. RESULTS: The recombinant RNASET2 was identified as a 27-kilodalton glycoprotein that possessed the ability to bind actin in vitro. RNASET2 significantly inhibited clonogenicity in HT29 cells. EI RNASET2 produced a similar effect, suggesting that its antitumorigenic activity is unrelated to its RNase activity. In HUVE cells, RNASET2 inhibited angiogenin-, bFGF-, and VEGF-induced tube formation in a dose-dependent manner. In athymic mice, RNASET2 inhibited the development of an LS174T-derived xenograft by 40%. A synergistic effect was obtained with combined RNASET2 and paclitaxel treatments. CONCLUSIONS: The current results suggested that RNASET2 represents a new class of antitumorigenic and antiangiogenic drugs, and the findings of this study emphasize the advantage of using agents like RNASET2 in combined therapy. PMID- 17109445 TI - Metastatic disease of screen-detected prostate cancer : characteristics at diagnosis. AB - BACKGROUND: Screening for prostate cancer has not only led to a stage migration, but also to a higher incidence of the disease. A decrease in mortality has occurred in several countries during the same time period. Risk stratification of screen-detected cancers at diagnosis has become more important for the anticipation and interpretation of changing incidence/mortality ratios. METHODS: From 1993 to 1998, 633 men were diagnosed with nonmetastatic prostate cancer in the prevalence screen of the Rotterdam section of the European Randomized study of Screening for Prostate Cancer (ERSPC). The characteristics at diagnosis of men who developed metastatic disease were compared with men without evidence of metastases during follow-up. RESULTS: During the median follow-up of 7.5 years, 41 men developed metastatic disease. After 10 years the metastasis-free survival rate was 89.6%, the overall survival 64.7%. In a Cox-model 2logPSA (prostate specific antigen), biopsy Gleason score and the number of biopsy cores with prostate cancer were independent predictors for the development of metastases; the latter only predicted metastases that presented within 60 months of follow up. CONCLUSIONS: The metastasis-free survival of men with prostate cancer detected in a prevalence screening was very high. Whether this was related to the beneficial effects of screening or to overdiagnosis due to screening (or both) remains unclear. The prognostic factors known for clinically diagnosed disease also hold for screen-detected disease. PMID- 17109446 TI - Phase II study of intravenous TZT-1027 in patients with advanced or metastatic soft-tissue sarcomas with prior exposure to anthracycline-based chemotherapy. AB - BACKGROUND: TZT-1027, a novel chemotherapeutic agent, is derived from dolastatin 10, and blocks cells during G2/M-phase by interfering with microtubule assembly and stability. TZT-1027 has exhibited potential cytotoxic activity in several human cancer cell lines (in vitro) and also demonstrated antitumor activity in human xenografts (in vivo). In addition, Phase I clinical investigations suggested activity in STS (soft-tissue sarcoma). METHODS: Eligible patients were those who had histologic evidence of locally advanced or metastatic STS and who had received 1 prior treatment regimen with an anthracycline-based chemotherapy for metastatic disease. Subjects received intravenous infusions of TZT-1027 over 1 hour on Day 1 and Day 8 of each 21-day treatment course. Efficacy was evaluated per Response Evaluation Criteria in Solid Tumors (RECIST) criteria. RESULTS: Twenty-nine patients were enrolled and 28 patients received at least 1 course of study drug and were eligible for efficacy and safety evaluation. The median age of the patients was 48 years (range, 23-73 years) and the median baseline Eastern Cooperative Oncology Group (ECOG) performance status was 1 (range, 0-2). A total of 67 courses (range, 1-9 courses; median, 2 courses) of TZT-1027 were administered. No patient in the study demonstrated an objective response to treatment. Of 6 patients (21.4%) who experienced disease stabilization, 1 continued to have stable disease for 9.3 months. The median time to tumor progression was 44 days (95% confidence interval [95% CI], 43.0-54.0) and the median survival was 178 days (95% CI, 134.0-317.0). The most commonly reported toxicities were neutropenia, fatigue, and constipation. CONCLUSIONS: TZT-1027 was found to be safe and well tolerated, and the hematologic toxicities observed were consistent with preclinical toxicology and Phase I study findings. No confirmed responses were seen in the current study. PMID- 17109447 TI - Cisplatin and gemcitabine with either vinorelbine or paclitaxel in the treatment of carcinomas of unknown primary site : results of an Italian multicenter, randomized, phase II study. AB - BACKGROUND: : To date, the standard treatment for patients who have carcinoma of unknown primary site has not been established. METHODS: : In this randomized Phase II study, 66 previously untreated patients (33 patients per arm) with carcinomas of unknown primary site received cisplatin (35 mg/m2) and gemcitabine (1000 mg/m2) with either paclitaxel (70 mg/m2) or vinorelbine (25 mg/m2), and all drugs were administered intravenously on Days 1 and 8 of a 21-day cycle. Twenty nine patients (44%) presented with > or =2 involved sites. The pathologic diagnosis was mainly adenocarcinoma (48 patients; 72.7%) and squamous carcinoma (7 patients; 10.6%). RESULTS: : In the first arm, 16 patients (48.5%) experienced an objective response, and 9 patients (27.2%) had disease stabilization. In the vinorelbine-containing arm, 14 patients (42.3%) experienced an objective response, and 8 patients (24.2%) had disease stabilization. The median response duration and the median time to progression were similar in both treatment arms; the median overall survival was 9.6 months (95% confidence interval, 7.11-12.09 months) for patients who received the cisplatin/gemcitabine/paclitaxel regimen and 13.6 months (95% confidence interval, 6.61-20.59 months) for patients who received the vinorelbine combination. Grade 3 and 4 toxicities were more frequent in the paclitaxel-containing arm. CONCLUSIONS: : Both combinations satisfied the 2-step design, demonstrating antitumor activity without relevant differences in response rates or response duration; however, the vinorelbine-containing regimen yielded superior results both in terms of overall survival (13.6 months vs 9.6 months) and in terms of treatment tolerability. Therefore, according to a pick the winner attitude, the combination of cisplatin/gemcitabine/vinorelbine may be considered in the design of future randomized, Phase III trials for patients with carcinomas of unknown primary site. PMID- 17109448 TI - Population-based study of renal cell carcinoma in children in Germany, 1980-2005: more frequently localized tumors and underlying disorders compared with adult counterparts. AB - BACKGROUND: Childhood renal cell carcinomas (RCCs) differ histologically and biologically from their adult counterparts. The characteristics of RCC-affected children and their tumors, the influence of treatment, and outcome have so far not been studied in a nonselected, population-based cohort. METHODS: A retrospective analysis was undertaken of RCC patients less than 16 years old reported to the German Childhood Cancer Registry and to the Kiel Paediatric Tumor Registry from 1980 to 2005. RESULTS: Forty-nine RCC in children (24 boys, 25 girls) with a median age of 10.6 years were identified. In about every third child possibly RCC-related underlying disorders (tuberous sclerosis, neuroblastoma, teratoma with chemotherapy, Saethre-Chotzen syndrome, chronic renal failure) or related diseases in their family were found. The pathologic subtypes were papillary in 16 (33%), translocation type in 11 (22%), unclassified in 8 (16%), and rarely clear-cell (n = 3) or others. Thirty-four (69%) patients had localized RCC, 8 (16%) patients regional lymph node metastases, and 4 (8%) patients distant metastases. Event-free survival and overall survival rates at 5 years were 96% for localized RCC, 69% and 75% for regional lymph node-positive, 25% and 33% for distant metastatic RCC, respectively. Two of 4 patients with distant metastases received immunotherapy combined with chemotherapy and surgery, both are alive, 1 of them disease-free for 6.9 years. CONCLUSIONS: Pediatric RCCs are predominantly localized diseases. Children with RCC frequently suffer underlying disorders. Survival rates in localized and regional lymph node positive cases are favorable. Because of the rarity of RCC in childhood, an international study is necessary. PMID- 17109449 TI - Estimation of the optimal brachytherapy utilization rate in the treatment of carcinoma of the uterine cervix: review of clinical practice guidelines and primary evidence. AB - BACKGROUND: Brachytherapy (BT) is an integral part of cervical carcinoma treatment. There have been no attempts to estimate the optimal proportion of new cervical carcinoma cases that should be treated with BT, that is, the optimal rate of brachytherapy utilization (BTU). METHODS: Evidence-based guidelines and primary evidence were used to construct a BTU tree for carcinoma of the uterine cervix. Searches were performed of the epidemiological literature to ascertain the proportion of patients who fulfilled criteria for BT. The robustness of the model was tested by sensitivity analyses and by peer review. A patterns of care study of BT in New South Wales for 2003 was conducted, and actual BTU for cervical carcinoma determined. The differences between optimal and actual rates of BTU were assessed. RESULTS: The optimal cervical carcinoma BTU was 49% (range, 42% to 50%). In New South Wales in 2003, actual BTU was only 30% of 256 cervical carcinoma patients. The major discrepancy was for FIGO stage IB-IIA disease, where there was an underutilization of BT, estimated to be 15% actual use compared with 47% optimal use. In Surveillance, Epidemiology, and End Results (SEER) areas, there was underutilization for stage IB-IIA (22% actual BTU versus 47% optimal BTU) and for stage IIB-IVA (54% actual BTU versus 100% optimal BTU). CONCLUSIONS: BT for cervical carcinoma is underutilized in New South Wales and in SEER areas. The authors' model of optimal BTU can be used as a quality assurance tool to provide an evidence-based benchmark against which actual patterns of practice can be measured. The model can also be used to help determine adequacy of BT resource allocation. PMID- 17109451 TI - A versatile ruthenium catalyst for C-C bond formation by C-H bond activation. PMID- 17109452 TI - Ice recrystallization inhibition and molecular recognition of ice faces by poly(vinyl alcohol). AB - The effects of poly(vinyl alcohol) (PVA) on the Ostwald ripening of polycrystalline ice samples are studied. At -6 degrees C, ice recrystallization in sucrose solutions is inhibited at PVA concentrations down to 0.005 mg mL(-1), with a recrystallization inhibition constant of 48.9 mL mg(-1). Ice growth-habit experiments reveal molecular recognition of the arrangement of water molecules in the ice by PVA molecules, and indicate that PVA molecules adsorb to the primary and secondary prism faces of hexagonal ice, Ih. Based on these observations, together with an analysis of the O-atom pattern in ice and the conformation of OH groups in PVA, an adsorption model is proposed. We suggest that PVA segments adsorb to the primary and secondary prism faces of ice parallel to the c axis with a linear misfit parameter of only 2.7 %, most likely via multiple hydrogen bonds. The proposed adsorption mechanism is discussed in the light of recent thermal hysteresis and scanning tunneling microscopy experiments. PMID- 17109453 TI - Synthesis and photoinduced electron-transfer process of a novel triphenylamine substituted polyfluorene-C60 triad. AB - The photoinduced electron-transfer process of a newly prepared, soluble, pi conjugated poly[9,9-bis(4-diphenylaminophenyl)-2,7-fluorene] (PDPAF), covalently bridged, C60 triad (C60-PDPAF-C60) is described. The molecular orbital calculations revealed that the majority of the highest occupied molecular orbital (HOMO) is located on the polyfluorene entity, while the lowest unoccupied molecular orbitals (LUMO) are found to be entirely on the C60 entity. The excited state electron-transfer processes were monitored by both steady-state and time resolved emission as well as by transient absorption techniques in toluene and benzonitrile. By excitation of the polyfluorene moiety, fluorescence quenching of the singlet excited state of polyfluorene moiety was observed. The nanosecond transient spectra in near-IR region revealed the charge-separation process from the polyfluorene moieties to the C60 moiety through the excited singlet states of polyfluorene. The lifetimes of the charge separated states were evaluated to be 20-50 ns, depending on the solvent polarity. PMID- 17109454 TI - Redox targeting of insulating electrode materials: a new approach to high-energy density batteries. PMID- 17109455 TI - Selective platination of modified oligonucleotides and duplex cross-links. PMID- 17109456 TI - Polyol synthesis through hydrocarbon oxidation: de novo synthesis of L-galactose. PMID- 17109457 TI - Fluoroproline flip-flop: regiochemical reversal of a stereoelectronic effect on peptide and protein structures. PMID- 17109458 TI - High-temperature-stable catalysts by hollow sphere encapsulation. PMID- 17109459 TI - Control of main-chain stiffness of a helical poly(phenylacetylene) by switching on and off the intramolecular hydrogen bonding through macromolecular helicity inversion. PMID- 17109460 TI - Comparison of activity of immobilised and soluble metalloporphyrins by NO release of oximes by chemiluminescence. AB - The ongoing interest in very efficient systems for the imitation of cytochrome P 450-dependent monooxygenase reactions, consisting of metalloporphyrin and oxygen donor, prompted us to develop a method to compare the catalytic activity of soluble metalloporphyrins with those which have been immobilised on different silica surfaces. In this article, we describe a method which allows direct comparison between the catalytic activities of soluble and immobilised metalloporphyrins by measuring the NO release of cyclohexanone oxime. We observed that all metalloporphyrins are able to enhance the NO release of cyclohexanone oxime in the presence of iodosobenzene as oxygen donor. In case of cationic metalloporphyrins, no difference in the catalytic activity was determined. In contrast, for perfluorinated metalloporphyrins we ascertained a major loss of activity after immobilisation. These results led us to the assumption that it is not easy to compare quantitative results of NO releases if soluble and immobilised metalloporphyrins were used. PMID- 17109461 TI - 3-heptylamino-5-phenylpyridazine derivatives as analogues of Acyl-CoA: cholesterol acyltransferase inhibitors containing the N-heptyl-N'-arylureidic moiety. AB - A series of novel Acyl-CoA: cholesterol acyltransferase (ACAT) inhibitors 8a-f was synthesized; the substances were characterized by the presence of a 2,5 dimethylpyrazin-3-yl moiety at one end and a 3-heptylamino-5-phenylpyridazine system at the other one, linked through linear alkyl spacers of different length. The new derivatives were designed based on the hypothesis that the 3-amino-5 phenylpyridazine moiety could mimic the aryl substituted urea, which was present in a number of ACAT inhibitors previously described. The choice of the 2,5 dimethylpyrazin-3-yl substituent was supported by a preliminary investigation, which indicated that this moiety is the most powerful in conferring ACAT inhibitory properties to the new series. The pharmacological results proved the idea to be sound. Finally, compounds 9a-c, lacking the phenylpyridazine moiety were prepared and tested to further strengthen our hypothesis. PMID- 17109462 TI - Synthesis, acute toxicity, and analgesic activity of new derivatives of pyrrole. AB - Ten pyrrole derivatives (including six new compounds) were synthesized and evaluated as potential platform for analgesic agents' development. Acute intraperitoneal toxicity and analgesic activity studies (acetic acid writhing test) were performed on mice with acetylsalicylic acid used as a reference substance. Products 3c, 3d, 3e, and 3h exhibited a dose-dependent activity demonstrating 1.5 to 2.5-fold better protections than the reference. The most prospective compounds comprised salicylic acid moieties, whose 4-substituted derivatives were related to lower acute toxicity and considerable activity. 4-[3 (Ethoxycarbonyl)-2-methyl-5-(3,4-dimethoxy-phenyl)-1H-pyrrol-1-yl]-2-hydroxy benzoic acid 3c was pointed out as the most prospective substance due to its lower acute toxicity (378 mg/kg body weight, intraperitoneally) and highest analgesic activity (up to 89.3% protection) in a dose range of 1/10 to 1/40 parts of LD(50). PMID- 17109463 TI - First total synthesis of the 2,7-naphthyridine alkaloids lophocladine a and B. AB - The one-pot reaction of 4-benzylpyridine-3-carbonitrile with Bredereck's reagent and subsequent treatment with either glacial acetic acid and sulfuric acid or ammonium acetate provided the new bioactive naphthyridine alkaloids lophocladine A and B, respectively. PMID- 17109464 TI - Src homology 2 domain-containing protein tyrosine phosphatase substrate 1 regulates the induction of Langerhans cell maturation. AB - Recently, we reported that Src homology 2 domain-containing protein tyrosine phosphatase substrate 1 (SHPS-1) plays an important role in the migration of Langerhans cells (LC). Here, we show that SHPS-1 is involved in the maturation of LC. Immunofluorescence analysis on epidermal sheets for I-A or CD86 revealed that LC maturation induced by 2,4-dinitro-1-fluorobenzene (DNFB) or by TNF-alpha was inhibited by pretreatment with an anti-SHPS-1 monoclonal antibody (mAb) or with CD47-Fc fusion protein, a ligand for SHPS-1. Further, FACS analysis demonstrated that I-A(+) LC that had emigrated from skin explants expressed CD80 or CD86, whereas CD47-Fc protein reduced CD80(high+) or CD86(high+) cells. CD47-Fc protein also reduced the up-regulation of surface CD80 or CD86 by LC remaining in the skin explants. In SHPS-1 mutant mice, we observed that the up-regulation of surface CD86 and CCR7 by LC induced by DNFB as well as that of surface CD80 and CD86 by LC in skin explants was attenuated. Finally, contact hypersensitivity (CHS) response was suppressed in SHPS-1 mutant mice and in wild-type mice treated with an anti-SHPS-1 mAb. These observations indicate that SHPS-1 plays an important role in the maturation of LC ex vivo and in vivo, and that SHPS-1-CD47 interaction may negatively regulate CHS. PMID- 17109465 TI - Inhibitory effect of the polyinosinic-polycytidylic acid/cationic liposome on the progression of murine B16F10 melanoma. AB - Cellular proteins, retinoic acid inducible gene-I and Toll-like receptor 3, sense dsRNA including polyinosinic-polycytidylic acid (PIC) to stimulate innate immune response. The local administration of PIC has been demonstrated to be effective in anti-tumor immunotherapy. However, the effects of PIC delivered cross the cell membrane have not yet been examined. To address this issue, we used a complex of PIC and cationic liposome (PIC liposome) and examined its anti-tumor effects in vitro and in vivo. PIC liposome could directly suppress the growth of B16F10 melanoma in vitro and repeated peritumoral injections of PIC liposome inhibited melanoma growth in a dose-dependent manner. This treatment induced tyrosinase related protein-2 (TRP-2)-tetramer(+) CD8(+) cells in the lymph nodes. As the mechanism for its anti-tumor immune response, we showed that the intradermal injection of PIC liposome induced the maturation of dendritic cells (DC). Moreover, the intratumoral injection of immature DC after treatment with PIC liposome significantly increased the number of TRP-2-specific IFN-gamma-producing cells in the lymph nodes as well as spleen, which resulted in an augmentation of the anti-tumor immune response. These studies demonstrate the potential of peritumoral injection of PIC liposome as immunotherapy for malignant melanoma. PMID- 17109466 TI - Immunosuppressive components of Ascaris suum down-regulate expression of costimulatory molecules and function of antigen-presenting cells via an IL-10 mediated mechanism. AB - High-molecular-weight components (PI) of Ascaris suum suppress both cell-mediated and humoral responses against ovalbumin (OVA) via an IL-4/IL-10-dependent mechanism. The aim of this work was to investigate the effect of PI on the ability of APC to activate T cells and the role of IL-10 in this process. Flow cytometry analyses of MHC class II, CD80, CD86 and CD40 molecules on LN cells from mice immunized with OVA or OVA+PI showed that PI inhibits expression of these molecules on unfractionated cells and on purified CD11c(+) cells. A low proliferative response was obtained when OVA-specific TCR-Tg T cells were incubated with CD11c(+) cells from OVA+PI-immunized mice pulsed with OVA, when compared to those incubated with cells from OVA-immunized mice. Similar results were obtained using as APC CD11c(+) cells from OVA-immunized mice pulsed with OVA+PI, which also expressed less of the four markers. The inhibitory effect of PI on both the expression of costimulatory molecules and the induction of T cell proliferation was abolished in IL-10-deficient mice. Our data indicate that the potent immunosuppressive effect of A. suum extract components on the host immune system is primarily related to their property of down-regulating the Ag presenting ability of DC via an IL-10-mediated mechanism. PMID- 17109467 TI - Alteration of epitope recognition pattern in Ag85B and ESAT-6 has a profound influence on vaccine-induced protection against Mycobacterium tuberculosis. AB - To analyze the effect of vaccine delivery systems on antigen recognition and vaccine efficacy, we compared immune responses in mice immunized either with an adenovirus vector expressing a fusion of Ag85B and ESAT-6 or with the recombinant fusion protein in a liposomal adjuvant. Both vaccines induced high levels of antigen-specific IFN-gamma production. The adjuvanted protein vaccine induced primarily a CD4 T cell response directed to the epitope Ag85B(241-255) and gave efficient protection against subsequent Mycobacterium tuberculosis infection. In contrast, the adenoviral construct induced a strong CD8 response predominantly targeted to the epitope ESAT-6(15-29) and no significant protection against infection. Vaccination with the protein vaccine resulted in highly accelerated recall of Ag85B(241-255)-specific T cells immediately post M. tuberculosis challenge whereas the ESAT-6(15-29) epitope was barely recognized during infection. Delivery of the viral construct in cationic liposomes switched the immune response to a protective one dominated by CD4 T cells targeted to the Ag85B(241-255) epitope. These data demonstrate that the nature of the T cell response to a vaccine antigen is more important than its magnitude with respect to protective efficacy and that vaccine-mediated changes in immunodominance can result in T cell responses of limited relevance during the natural infection. PMID- 17109468 TI - CD4+ T cells specific for a model latency-associated antigen fail to control a gammaherpesvirus in vivo. AB - CD4(+) T cells play a major role in containing herpesvirus infections. However, their cellular targets remain poorly defined. In vitro CD4(+) T cells have been reported to kill B cells that harbor a latent gammaherpesvirus. We used the B cell-tropic murine gammaherpesvirus-68 (MHV-68) to test whether this also occurred in vivo. MHV-68 that expressed cytoplasmic ovalbumin (OVA) in tandem with its episome maintenance protein, ORF73, stimulated CD8(+) T cells specific for the H2-K(b)-restricted OVA epitope SIINFEKL and was rapidly eliminated from C57BL/6 (H2(b)) mice. However, the same virus failed to stimulate CD4(+) T cells specific for the I-A(d)/I-A(b)-restricted OVA(323-339) epitope. We overcame any barrier to the MHC class II-restricted presentation of an endogenous epitope by substituting OVA(323-339) for the CLIP peptide of the invariant chain (ORF73-IRES Ii-OVA), again expressed in tandem with ORF73. This virus presented OVA(323-339) but showed little or no latency deficit in either BALB/c (H2(d)) or C57BL/6 mice. Latent antigen-specific CD4(+) T cells therefore either failed to recognize key virus-infected cell populations in vivo or lacked the effector functions required to control them. PMID- 17109469 TI - The structure of the human allo-ligand HLA-B*3501 in complex with a cytochrome p450 peptide: steric hindrance influences TCR allo-recognition. AB - Virus-specific T cell populations have been implicated in allo-recognition. The subdominant T cell receptor JL12 recognizes both HLA-B*0801 presenting the Epstein-Barr virus-derived peptide FLRGRAYGL and also HLA-B*3501 presenting the cytochrome p450 self peptide KPIVVLHGY. This cross-reactivity could promote the rejection of HLA-B*3501-positive cells in Epstein-Barr virus-exposed HLA-B*0801 recipients. LC13, the dominant TCR against the HLA-B*0801:FLRGRAYGL complex, fails to recognize HLA-B*3501:KPIVVLHGY. We report the 1.75-Angstrom resolution crystal structure of the human allo-ligand HLA-B*3501:KPIVVLHGY. Similarities between this structure and that of HLA-B*0801:FLRGRAYGL may facilitate cross recognition by JL12. Moreover, the elevated peptide position in HLA B*3501:KPIVVLHGY would provide steric hindrance to LC13, preventing it from interacting in the manner in which it interacts with HLA-B*0801:FLRGRAYGL. These findings are relevant to understanding the basis of T cell cross-reactivity in allo-recognition, optimal transplant donor-recipient matching and developing specific molecular inhibitors of allo-recognition. PMID- 17109470 TI - NF-kappa B binds to the immunoglobulin S gamma 3 region in vivo during class switch recombination. AB - Ig class switch recombination (CSR) is dependent upon the expression of activation-induced deaminase and targeted to specific isotypes by germ-line transcript expression and isotype-specific factors. NF-kappaB plays critical roles in multiple aspects of B cell biology and has been implicated in the mechanism of CSR by in vitro binding assays and altered S/S junctions derived from NF-kappaB p50-deficient mice. However, the pleiotropic contributions of NF kappaB to gene expression in B cells has made discerning a direct role for NF kappaB in CSR difficult. We now observe that binding of NF-kappaB components p50 and p65 is detected on Sgamma3 in vivo following lipopolysaccharide (LPS) activation and repressed by LPS + IL-4, suggesting a direct role for this factor in CSR. In vivo footprinting confirms occupancy of a previously defined NF-kappaB recognition site in Sgamma3 with the same temporal kinetics as found in the chromatin immunoprecipitation analysis. Binding of NF-kappaB components p50 and p65 was also detected on Sgamma1 following B cell activation. H3 histone hyper acetylation at Sgamma1 is strongly correlated with NF-kappaB binding, suggesting that NF-kappaB mediates chromatin remodeling in the Sgamma3 and Sgamma1 region. PMID- 17109471 TI - Immunomodulatory effects associated with a live vaccine against Leishmania major containing CpG oligodeoxynucleotides. AB - The inoculation of live Leishmania major to produce a lesion that heals (leishmanization) is to date the only vaccine against cutaneous leishmaniasis that has proven effective in humans, but it still has an unacceptable frequency of large ulcerating lesions that are slow to heal or, in rare cases, non-healing. We have previously shown that C57BL/6 mice vaccinated intradermally with 10(4) L. major/50 microg CpG oligodeoxynucleotides develop little or no dermal lesions and show early containment of parasite growth in the vaccination site, eliminating safety concerns related to the inoculation of live organisms. The addition of CpG to the live vaccine resulted in early activation of dermal dendritic cells and increased IL-6 production, as well as in a reduction in the accumulation of Foxp3(+)CD4(+)CD25(+) regulatory T (T(reg)) cells that naturally occurs in the skin following Leishmania infection. Neutralization of IL-6 caused the development of larger lesions and increased local T(reg) cell numbers. Transfer of vaccine-primed dendritic cells into IL-6-deficient mice mitigated lesion development, indicating that IL-6 reconstitution limited pathology in the vaccination site. PMID- 17109472 TI - Activation-induced cell death of human melanoma specific cytotoxic T lymphocytes is mediated by apoptosis-inducing factor. AB - Activation-induced cell death (AICD) of T cells can be an impediment towards achieving a robust and long-lived cytolytic T lymphocyte (CTL) response from active specific immunization or after adoptive cell transfer in cancer immunotherapy. The mechanism of AICD in primary CTL, however, remains poorly understood. It is widely believed that AICD is driven by signals from death receptors (DR) and that the cell death takes place in a caspase-dependent manner, although it has been shown that AICD of T cells can be induced by internal triggers and that death takes place in a caspase-independent manner. We show here that AICD in human melanoma epitope-specific primary CTL involves selective mitochondrio-nuclear translocation of the apoptosis inducing factor (AIF) without cytochrome c release, caspase-3 and caspase-8 activation, and results from large scale DNA fragmentation. The c-jun-N terminal kinase (JNK) inhibitor, SP600125, blocks the mitochondrio-nuclear translocation of AIF and prevents AICD in these CTL. These findings suggest that the AICD in human melanoma epitope specific primary CTL is mediated by mitochondrial AIF release and JNK is involved in regulation of this death process. PMID- 17109473 TI - Expression and function of NKG2D in CD4+ T cells specific for human cytomegalovirus. AB - The human NKG2D killer lectin-like receptor (KLR) is coupled by the DAP10 adapter to phosphoinositide 3-kinase (PI3 K) and specifically interacts with different stress-inducible molecules (i.e. MICA, MICB, ULBP) displayed by some tumour and virus-infected cells. This KLR is commonly expressed by human NK cells as well as TCRgammadelta(+) and TCRalphabeta(+)CD8(+) T lymphocytes, but it has been also detected in CD4(+) T cells from rheumatoid arthritis and cancer patients. In the present study, we analysed NKG2D expression in human cytomegalovirus (HCMV) specific CD4(+) T lymphocytes. In vitro stimulation of peripheral blood mononuclear cells (PBMC) from healthy seropositive individuals with HCMV promoted variable expansion of CD4(+)NKG2D(+) T lymphocytes that coexpressed perforin. NKG2D was detected in CD28(-) and CD28(dull )subsets and was not systematically associated with the expression of other NK cell receptors (i.e. KIR, CD94/NKG2 and ILT2). Engagement of NKG2D with specific mAb synergized with TCR-dependent activation of CD4(+) T cells, triggering proliferation and cytokine production (i.e. IFN-gamma and TNF-alpha). Altogether, the data support the notion that NKG2D functions as a prototypic costimulatory receptor in a subset of HCMV specific CD4(+) T lymphocytes and thus may have a role in the response against infected HLA class II(+) cells displaying NKG2D ligands. PMID- 17109474 TI - U-500 insulin: why, when and how to use in clinical practice. AB - Some patients with type 2 diabetes mellitus (T2DM) have severe insulin resistance. Their insulin requirements are significantly greater. These patients need to take 2-3 injections at the same time to take the correct insulin dose or to redial the insulin pen. When daily insulin requirements are in excess of 300 units/day, the volume of the injected insulin becomes an issue. Large-volume injection can cause discomfort and lead to poor concordance with treatment. Using high-strength insulin e.g. U-500 insulin can reduce the volume of the injected insulin. Despite publications of small case reports or case series, no universal guidelines exist on the use of U-500 insulin. We discuss common sense approaches when considering the use of U-500 insulin in clinical practice. PMID- 17109475 TI - Slow versus fast proteins in the stimulation of beta-cell response and the activation of the entero-insular axis in type 2 diabetes. AB - BACKGROUND: We tested whether ingestion of whey protein can induce greater post prandial amino acid (AA) levels in the plasma and a higher beta-cell response than casein ingestion in type 2 diabetes mellitus patients. METHODS: The study was designed as a double-blind, randomized, and controlled cross-over clinical trial. Twelve post-absorptive type 2 diabetic subjects who were withdrawn from their usual hypoglycemic therapy were studied. A medium calorie (approximately 6 kcal/kg BW), high protein (approximately 50% of total kcal) mixed meal, containing whey protein, casein, or a free amino acid (FREE AA) mixture matching the casein AA composition, was randomly administered on three different occasions. RESULTS: Following ingestion of whey protein, plasma concentrations of total, branched chain, and essential AA were 25-50% greater than after ingestion of casein (p < 0.0001), and were similar to those observed after the FREE AA meal. With whey protein, C-peptide, insulin, and pro-insulin concentrations were greater by 12-40% (p < 0.02 or less) than with casein, and similar to those with FREE AA. Glucagon-like polypeptide 1 (GLP-1) response tended to be lower with casein than with whey protein. Glucose-dependent insulinotropic polypeptide (GIP) response was greater with either whey protein or casein than with FREE AA. Post prandial glucose concentrations were similar after whey protein and casein ingestion, but lower after the FREE AA meal. CONCLUSIONS: In type 2 diabetes, the ingestion of a fast-absorbable protein results in a greater post-prandial aminoacidemia and a higher beta-cell secretion than the ingestion of a 'slow' protein. Whether these changes can be maintained chronically in combination with hypoglycemic therapy, possibly also resulting in better glycemic control, remains to be established. PMID- 17109476 TI - Identification of responders and reactive domains to rivastigmine in Alzheimer's disease. AB - PURPOSE: Presently, it is unclear which patients suffering from Alzheimer's Disease (AD) respond to rivastigmine and if rivastigmine acts on specific cognitive domains. The aims of this study are thus to investigate treatment effects of rivastigmine on specific cognitive domains and to find possible responsive subpopulations to rivastigmine cognitive effects. METHODS: Mini Mental State Examination (MMSE) and Cambridge Cognitive Examination (CAMCOG) were administered at baseline and after 6 months in 83 rivastigmine users and 96 historical controls, representing natural decline. Treatment effects on different subsections of the CAMCOG and in different subpopulations were investigated by linear regression analyses. RESULTS: Rivastigmine showed effectiveness on total CAMCOG (p < 0.001), CAMCOG non-memory subsection (p < 0.001) and subscales of language (p = 0.002), attention/calculation (p = 0.043), abstract thinking (p < 0.001) and perception (p = 0.031). In patients with baseline MMSE < or =19 rivastigmine showed significant and favourable effects compared to historical controls on total CAMCOG (p < 0.001) and both non-memory (p < 0.001) and memory subsections (p = 0.002). CONCLUSION: Rivastigmine showed primarily effectiveness on the non-memory section of the CAMCOG and patients with a baseline MMSE < or = 19 appeared to show greater responses to rivastigmine compared to patients with baseline MMSE > or = 20. PMID- 17109483 TI - Directed evolution of the 5'-untranslated region of the phoA gene in Escherichia coli simultaneously yields a stronger promoter and a stronger Shine-Dalgarno sequence. AB - Directed evolution has been widely applied for gene improvement through random mutagenesis of coding sequences. Through error-prone PCR both in the coding sequence and the regulatory sequence of E. coli alkaline phosphatase, the cellular enzyme activity has been efficiently enhanced. Sequence analysis revealed that the resultant mutant 34-B12, which showed a sevenfold increased enzyme activity at the cellular level, contains three mutations in the regulatory sequence and another three mutations in the coding sequence. Activity assays of the enzyme containing the corresponding amino acid substitutions proved that the amino acid mutations contribute only to a small portion to the increased cellular enzyme activity. So the mutations in the 5'-untranslated region were analyzed separately and combinationally. The results suggested that one mutation yielded a stronger promoter and the other two mutations both elevated the E. coli alkaline phosphatase expression at the translational level; moreover, a stronger Shine Dalgarno sequence was generated. PMID- 17109484 TI - Precipitants and additives for membrane crystallization of lysozyme. AB - Membrane crystallization is a newly developed crystallization technique that has proven to be superior in producing good crystal forms under operating conditions that are not appropriate to perform the crystallization process by other traditional techniques. In this work, static membrane crystallization was carried out on lysozyme, with hollow-fiber microporous hydrophobic membranes. Numerous precipitant and additive types and concentrations were employed in the crystallization processes in order to select the most appropriate precipitant and additive types and to find their corresponding concentration levels that can yield the best crystal forms. The crystallization processes were analyzed in two ways: firstly, by evaluation of the transmembrane fluxes obtained by using different precipitants and additives; secondly, by utilization of the images and results obtained from the micrography and IR spectra in comparisons and evaluations of the crystals formed under all kinds of conditions. Moreover, the size distributions of the crystals yielded under several typical crystallization conditions were analyzed, and turbidity and induction time periods obtained during typical crystallization experiments were also measured. Amongst the numerous precipitants and additives tested, the most appropriate precipitant type and additive were chosen and their concentrations were optimized. Good lysozyme crystals were obtained using a certain precipitant and additive. The obtained results from this work further support the advantages of utilizing the membrane crystallization technique for macromolecule crystallizations. PMID- 17109489 TI - Environmental biotechnology in China: progress and prospect. AB - Biotechnology could be widely applied in the technological development of environmental protection. This report gives a brief Chinese review on the major progress of environmental biotechnology in the following fields: Monitoring technology and treatment of dioxin-like chemicals and cyanobacterial toxins, biofiltration for air pollution control, solid waste treatment and reutilization, bioremediation of soil pollution, risk assessment and control of endocrine disturbing substances in environment, wastewater treatment, clean production and recycling economy. The effect of government policy and fund on the progress of environmental biotechnology is also discussed. PMID- 17109490 TI - BIOCENTURY, outrunner of China's biological technology in agriculture Biocentury Transgene (China) Co., Ltd. PMID- 17109491 TI - Biotechnology and bioeconomy in China. AB - From the review of the achievements and advantages in the development of biotechnology (BT) and bioindustry in China, it is clear that the bioeconomy would provide a tremendous opportunity for China to develop sustainably or even surpass a few developed countries. A long-term vision has been made to guide the research and development and industrialization of BT in China. This review detailed the strategies, targets, priorities, and key technologies in each stage. Furthermore, the reviewers expatiated on the establishment of the favorable policies, the foundation of the professional groups, the establishment of the advanced laboratories or centers, the investment mechanisms, the development and evaluation of biosafety, the encouragement and support for the international collaborations and exchanges, and the establishment of the general organizational structure. PMID- 17109492 TI - Development of a vaccine marker technology: display of B cell epitopes on the surface of recombinant polyomavirus-like pentamers and capsoids induces peptide specific antibodies in piglets after vaccination. AB - Highly immunogenic capsomers (pentamers) and virus-like particles (VLPs) were generated through insertion of foreign B cell epitopes into the surface-exposed loops of the VP1 protein of murine polyomavirus and via heterologous expression of the recombinant fusion proteins in E. coli. Usually, complex proteins like the keyhole limpet hemocyanin (KLH) act as standard carrier devices for the display of such immunogenic peptides after chemical linkage. Here, a comparative analysis revealed that antibody responses raised against the carrier entities, KLH or VP1 pentamers, did not significantly differ up to 18 weeks, demonstrating the highly immunogenic nature of VP1-based particulate structures. The carrier-specific antibody response was reproducibly detected in the meat juice after processing. More importantly, chimeric VP1 pentamers and VLPs carrying peptides of 12 and 14 amino acids in length, inserted into the BC2 loop, induced a strong and long lasting humoral immune response against VP1 and the inserted foreign epitope. Remarkably, the epitope-specific antibody response was only moderately decreased when VP1 pentamers were used instead of VLPs. In conclusion, we identified polyomavirus VP1-based structures displaying surface-exposed immunodominant B cell epitopes as being an efficient carrier system for the induction of potent peptide-specific antibodies. The application of this approach in vaccine marker technology in livestock holding and the meat production chain is discussed. PMID- 17109493 TI - Caenorhabditis elegans: a versatile platform for drug discovery. AB - Drug discovery and drug target identification are two intimately linked facets of intervention strategies aimed at effectively combating pathological conditions in humans. Simple model organisms provide attractive platforms for devising and streamlining efficient drug discovery and drug target identification methodologies. The nematode worm Caenorhabditis elegans has emerged as a particularly convenient and versatile tool that can be exploited to achieve these goals. Although C. elegans is a relatively modern addition to the arsenal of model organisms, its biology has already been investigated to an exceptional level. This, coupled with effortless handling and a notable low cost of cultivation and maintenance, allows seamless implementation of high-throughput drug screening approaches as well as in-depth genetic and biochemical studies of the molecular pathways targeted by specific drugs. In this review, we introduce C. elegans as a model organism with significant advantages toward the identification of molecular drug targets. In addition, we discuss the value of the worm in the development of drug screening and drug evaluation protocols. The unique features of C. elegans, which greatly facilitate drug studies, hold promise for both deciphering disease pathogenesis and formulating educated and effective therapeutic interventions. PMID- 17109494 TI - Distress overlaps with anxiety and depression in patients with head and neck cancer. AB - Psychological problems in cancer patients often go unrecognized until they are specifically sought. This is more in patients with depression as they are reluctant to complain about their symptoms. The present study was carried out to evaluate the relation of distress with anxiety and depression in 123 patients with head and neck cancers using Distress Inventory for Cancer version 2 (DIC2) and the Hospital Anxiety and Depression scale (HADS). The mean DIC 2 scores were 24.6 while that of subscales ranged from 2.6 to 11.0. Fifteen patients were found to have clinical caseness for anxiety while 12 (10%) were caseness for depression. Total distress, emotional and social distress subscales were found to have positive correlation with anxiety and depression suggesting a possible overlap of two constructs. In multivariate analysis only belief in god was found to significantly affect the distress. Results of present study suggest significant psychological morbidity in head neck cancer patients undergoing curative treatment. This is the first study reporting on the psychometric properties of distress inventory on cancer version 2 since its validation, the results suggest a possible overlap of two constructs similar to that seen with other tools on distress and this may have major implications for clinical practice. PMID- 17109495 TI - mRNA expression, functional profiling and multivariate classification of colon biopsy specimen by cDNA overall glass microarray. AB - AIM: To understand the local pathophysiological alterations and gene ontology based functional classification of colonic biopsies into inflammatory and neoplastic diseases. METHODS: Total RNA was extracted from frozen biopsies and amplified by T7-method. Expression profile was evaluated by Atlas Glass 1K microarrays. After microarray quality control, applicable data were available from 10 adenomas, 6 colorectal adenocarcinomas (CRCs), and 6 inflammatory bowel diseases (IBDs). Multivariate statistical and cell functional analyses were performed. Real-time RT-PCR and immunohistochemistry were used for validation. RESULTS: Discriminant analysis of selected genes, could correctly reclassify all 22 samples using 4 parameters (heat shock transcription factor-1, bystin-like, calgranulin-A, TRAIL receptor 3). IBD samples were characterized by overregulated chemokine (C-X-C motif) ligand 13, replication protein A1, E74-like factor 2 and downregulated TNF receptor-associated factor 6, BCL2-interacting killer genes. In adenomas upregulation of TNF receptor-associated factor 6, replication protein A1, E74-like factor 2 and underexpression of BCL2-associated X protein, calgranulin-A genes were found. CRC cases had significantly increased epidermal growth factor receptor, topoisomerase-1, v-jun, TNF receptor-associated factor 6 and TRAIL receptor 3, and decreased RAD51 and RAD52 DNA repair gene, protein phosphatase-2A and BCL2-interacting killer mRNA levels. Epidermal growth factor receptor RT-PCR and immunohistochemistry, topoisomerase-1 RT-PCR confirmed the chip results. CONCLUSION: Different histological alterations can be reclassified by functional, multivariate analysis using cDNA microarrays. Further studies with expanded sample number are needed for subclassification of pathological alterations. PMID- 17109496 TI - Colorectal cancer screening by non-invasive metabolic biomarker fecal tumor M2 PK. AB - AIM: To evaluate the utility of the innovative fecal tumor M2-Pyruvate kinase (M2 PK) test in our daily clinical routine, as a marker for the pre-selection of patients who should subsequently undergo colonoscopy for the diagnosis or exclusion of colorectal cancer. METHODS: Fecal tumor M2-PK was measured in stool samples of 96 study participants (33 patients with colorectal cancer, 21 patients with rectal carcinoma and 42 controls) who all underwent total colonoscopy. RESULTS: In 39 of 42 individuals in the control group, fecal tumor M2-PK was below 4.0 kU/L (93% specificity). Colorectal tumors were accompanied by a highly significant increase (P < 0.001) in fecal tumor M2-PK levels (median: colon carcinoma, 23.1 kU/L; rectal carcinoma, 6.9 kU/L; colorectal carcinoma, 14.7 kU/L), which correlated with Duke's staging and T-classification. The overall sensitivity was 78% for colorectal cancer, increasing from 60% for stage T1 to 100% for stage T4 and from 60% for Duke's A to 90% for Duke's D tumors. CONCLUSION: Fecal tumor M2-PK is an appropriately sensitive tool to pre-select those patients requiring colonoscopy for the further diagnostic confirmation or exclusion of colorectal cancer. PMID- 17109497 TI - Steroids reduce local inflammatory mediator secretion and mucosal permeability in collagenous colitis patients. AB - AIM: To study the effect of oral steroids upon clinical response and rectal mucosa secretion of eosinophil cationic protein (ECP), myeloperoxidase (MPO), basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF) and albumin in patients with collagenous colitis (CC). METHODS: A segmental perfusion technique was used to collect perfusates from rectum of CC patients once before and twice (one and four weeks) after the start of steroid treatment. Clinical data was monitored and ECP, MPO, bFGF, VEGF and albumin concentrations were analyzed by immunochemical methods in perfusates and in serum. RESULTS: Steroids reduced the number of bowel movements by more than five times within one week and all patients reported improved subjective well-being at wk 1 and 4. At the same time, the median concentrations of ECP, bFGF, VEGF and albumin in rectal perfusates decreased significantly. MPO values were above the detection limit in only 3 patients before treatment and in none during treatment. VEGF, bFGF, ECP and albumin concentrations correlated with each other with the exception of ECP and albumin. A decrease of serum ECP and VEGF concentrations was also seen even if the overtime reduction was not significant. CONCLUSION: Oral steroid treatment in CC patients induced a simultaneous reduction of bowel movements and rectal release of ECP, bFGF, VEGF and albumin, suggesting that these polypeptides and increased mucosal permeability are important components of the pathophysiology in collagenous colitis. PMID- 17109498 TI - Hepatitis B genotypes: relation to clinical outcome in patients with chronic hepatitis B in Saudi Arabia. AB - AIM: To identify the most common hepatitis B virus (HBV) genotype in Saudi Arabia, and correlate the prevailing genotypes with the clinical outcome of patients. METHODS: Patients were consecutively recruited from the hepatology clinics of two tertiary care referral centers. Patients were categorized into 4 different groups: group 1, patients with hepatitis B and normal liver enzymes; group 2, patients with hepatitis B and abnormal liver enzymes but without cirrhosis; group 3, patients with hepatitis B and liver cirrhosis; group 4, patients with hepatitis B and hepatocellular carcinoma. All patients had a positive hepatitis B surface antigen (HBsAg). Genotyping of HBV was performed by nested PCR-mediated amplification of the target sequence and hybridization with sequence-specific oligonucleotides. RESULTS: Seventy patients were enrolled in this study. They were predominantly male (72.9%) in their mid-forty's (mean age 47 years). Forty-nine (70%) patients were hepatitis B envelope antigen (HBeAg) negative. The majority of patients (64%) acquired HBV through unknown risk factors. Hepatitis B genotyping revealed that 57 patients (81.4%) were genotype D, 1 patient (1.4%) had genotype A, 1 patient (1.4%) had genotype C, and 4 patients (5.7%) had genotype E, while 7 patients (10%) had mixed genotype (4 patients ADG, 1 patient DE, 1 patient DF, and 1 patient ADFG). Based on univariate analysis of genotype D patients, significant predictors of advanced liver disease were age, gender, aspartate transaminase, alanine transaminase, albumin, bilirubin, and alkaline phosphatase (all P < 0.001). In multivariate analysis decreased hemoglobin (r = -0.05; 95% CI: -0.08 to -0.03; P = 0.001) and albumin levels (r = -0.004; 95% CI: -0.007 to -0.001; P = 0.002) were highly significant predictors of advanced liver disease. In patients with HBV genotype D, HBeAg negativity was found to increase across advancing stages of liver disease (P = 0.024). CONCLUSION: This study highlights that the vast majority of Saudi patients with chronic hepatitis B have genotype D. No correlation could be observed between the different genotypes and epidemiological or clinical factors. The relationship between genotype D and HBeAg status in terms of disease severity needs to be further elucidated in larger longitudinal studies. PMID- 17109499 TI - Hepatis C virus transmission and its risk factors within families of patients infected with hepatitis C virus in southern Iran: Khuzestan. AB - AIM: To determine whether hepatitis C virus (HCV) infection of index cases increases intrafamilial transmission (sexual and nonsexual contacts) of HCV. METHODS: In a case-control descriptive study we enrolled 300-household contacts of 60 index cases (40 males and 20 females) of HCV infection and 360 pair-matched controls in Ahwaz JundiShapour University Hospitals from August 1, 1998 to September 1, 2003. The control group consisted of first time blood donors referred to the Regional Blood Transfusion Organization. Serum samples and demographic data and a medical history including the existence of risk factors for HCV (after a questionnaire on the risk factors for parenteral exposure) were obtained from each subject. Antibodies to HCV were detected employing a commercially available second-generation enzyme immunoassay (EIA, Abbott II). Positive serum specimens were retested using a second-generation recombinant immunoblot assay (RIBA-2) and a polymerase chain reaction for HCV RNA. Data analysis was carried out for intra-household clustering. RESULTS: Only 4 of 300 (1.33%) cases of household contacts without percutaneous risk factors were positive for HCV Ab while the remaining 296 family contacts were negative for anti-HCV. The mean age of the index cases was 28.4 (Std 15.22) years. The anti HCV prevalences in parents, spouses, children of the index cases were 0.87% (1/115), 3.39% (2/59)) and 0.79% (1/126), respectively. Among couple partners negative for anti-HCV antibodies, the mean duration of the sexual relationship was 6 years. The two-couple partners positive for anti-HCV antibodies married the index cases for longer than 15 years. The prevalence of positive HCV Ab among household contacts (1.33%) was not significantly higher than that in the controls (1%) (P > 0.06). CONCLUSION: Intrafamilial transmission of HCV is not the significant transmission route and sexual transmission does not seem to play a role in the intrafamilial spread of HCV infection. Intrafamilial transmission of HCV is possible but occurs at a low rate. PMID- 17109500 TI - Evaluation of the role of H pylori infection in pathogenesis of gastric cancer by immunoblot assay. AB - AIM: To elucidate the different serological reactions to H pylori using the immunoblotting technique for further understanding of its pathogenic role in gastric cancer. METHODS: A total of 54 patients were divided into two groups after upper gastrointestinal endoscopy: normal control group (25 patients) and gastric cancer group (29 patients). Both groups were further divided into H pylori (+) and H pylori (-) subgroups based on the results of CLO test, Giemsa staining and culture. Sera were further analyzed with the immunoblotting technique (HelicoBlot 2.0, Genelabs Diagnostics, Singapore). RESULTS: The positive rate of the immunoblotting test was as high as 88.9% in the H pylori (-) gastric cancer group and only 14.3% in the H pylori (-) normal control group with a statistically significant difference. CONCLUSION: The prevalence of H pylori infection is higher in gastric cancer patients than in the normal controls, suggesting that H pylori may play a role in the pathogenesis of gastric cancer. PMID- 17109501 TI - Evaluation of intraoperative radiotherapy for gastric carcinoma with D2 and D3 surgical resection. AB - AIM: To study the proper sites and doses of intraoperative radiotherapy (IORT) for gastric carcinoma and the effects of this treatment. METHODS: A total of 106 patients with stage I-IV gastric carcinoma who received D2 or D3 radical operation combined with IORT were analyzed. Sixty-seven patients with gastric cancer of the antrum and body underwent distal gastrectomy. The sites of irradiation were at the celiac artery and hepatoduodenal ligment area. Another 39 patients with carcinoma of the cardia and upper part of the gastric body and whole stomach underwent proximal gastrectomy or total gastrectomy. The sites of irradiation for this group were the upper margin of the pancreas and the regional para-aorta. The therapeutic effects (including survival and complications) of these 106 cases received operation combined with IORT (IORT group) were compared with 441 cases treated during the same time period by a radical operation alone (operation group). RESULTS: The radiation dose below 30 Gy was safe. The therapeutic method of the operation combined with IORT did not prolong the survival of patients with stage I and IV gastric cancer, but the 5-year survival rates of patients with stage II and III gastric cancers were significantly improved. The 5-year survival rates of the stages III cancer patients receiving D2 resection combined with IORT were markedly improved, while for those receiving D3 radical resection, only the postoperative 3- or 4-year survival rates were improved (P < 0.005-0.001). The 5-year survival rate for those patients was raised only by 4.7% (P > 0.05). CONCLUSION: The 5-year survival rates of patients with stages II and III gastric carcinoma who received D2 lymphadenectomy combined with IORT were improved, and there was no influence on the postoperative complications and mortality. PMID- 17109502 TI - Alterations of biliary biochemical constituents and cytokines in infantile hepatitis syndrome. AB - AIM: To investigate the biliary biochemical constituents and cytokines in infantile hepatitis syndrome (IHS). METHODS: From 42 IHS subjects and 21 controls, serum and biliary biochemical constituents, including total bilirubin (TBIL), direct bilirubin (DBIL), alanine aminotransferase (ALT), gamma-glutamyl transpeptidase (gamma-GT), total bile acid (TBA), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) both in bile and serum, were assayed. The subjects with IHS were divided into a cholestasis group (n = 21) and a hepatitis group (n = 21). RESULTS: In the cholestasis group, serum TBIL, DBIL, ALT, gamma GT, TBA, IL-6 and TNF-alpha levels were higher than those in the control (P < 0.01); and also the biliary TBIL, DBIL, gamma-GT and TBA levels were lower than those in the control, whereas biliary IL-6 and TNF-alpha levels were higher than those in the control (P < 0.01). In the cholestasis group, serum IL-6 and TNF alpha levels were lower than those in bile (P < 0.01). In the hepatitis group, serum DBIL, ALT, gamma-GT, TBA, IL-6 and TNF-alpha levels were higher than those in the control (P < 0.01 or 140.57 +/- 70.32 vs 79.06 +/- 35.25, P < 0.05), while biliary TBIL, DBIL, gamma-GT and TBA levels were lower than those in the control (P < 0.01), and biliary IL-6 and TNF-alpha levels were higher than those in the control (P < 0.01). In the hepatitis group, serum IL-6 and TNF-alpha levels were also lower than those in bile (P < 0.01). Serum TBIL, DBIL, gamma-GT, IL-6 and TNF-alpha levels in the cholestasis group were higher than those in the hepatitis group, while biliary IL-6 and TNF-alpha levels in the cholestasis group were higher than those in the hepatitis group. Biliary IL-6 and TNF-alpha were found to be more significantly increased than serum IL-6 and TNF-alpha in IHS (P < 0.01). The biliary IL-6 and TNF-alpha levels were positively correlated with serum DBIL, TBA and gamma-GT levels in IHS subjects. CONCLUSION: Biliary biochemical constituents alter in coincidence with pathological changes in hepatocellular injury. Cholestasis is more serious in IHS patients of cholestasis subtype. Assay of biliary IL-6 and TNF-alpha levels can be specific and sensitive to determine the inflammatory status of impaired liver in IHS. PMID- 17109503 TI - Construction of an oral recombinant DNA vaccine from H pylori neutrophil activating protein and its immunogenicity. AB - AIM: To construct a live attenuated Salmonella typhimurium (S. typhimurium) strain harboring the H pylori neutrophil activating protein (HP-NAP) gene as an oral recombinant DNA vaccine, and to evaluate its immunogenicity. METHODS: By genetic engineering methods, the genomic DNA of H pylori was extracted as a template. The total length of the HP-NAP gene was amplified by polymerase chain reaction (PCR) and cloned into pBT vector for sequencing and BLAST analysis, then subcloned into a eukaryotic expression vector pIRES followed by PCR identification and restriction enzyme digestion. The identified recombinant plasmid pIRES-NAP was transfected into COS-7 cells for target fusion protein expression, and its antigenicity was detected by Western blotting. Then the recombinant plasmid was transformed into a live attenuated S. typhimurium strain SL7207 as an oral vaccine strain, and its immunogenicity was evaluated with animal experiments. RESULTS: A 435 bp product was cloned using high homology with HP-NAP gene in GenBank (more than 98%). With identification by PCR and restriction enzyme digestion, a recombinant eukaryotic expression plasmid pIRES NAP containing the HP-NAP gene of H pylori was successfully constructed. The expressed target protein had a specific reaction with H pylorii whole cell antibody and showed a single strip result detected by Western blotting. Oral immunization of mice with recombinant DNA vaccine strain SL7207 (pIRES-NAP) also induced a specific immune response. CONCLUSION: The successful construction of HP NAP oral DNA vaccine with good immunogenicity may help to further investigate its immunoprotection effects and develop vaccine against H pylori infection. PMID- 17109504 TI - Randomized clinical trial on seven-day-per-week continuous accelerated irradiation for patients with esophageal carcinoma: preliminary report on tumor response and acute toxicity. AB - AIM: Tumor response and normal tissue toxicity of seven-day-per-week continuous accelerated irradiation (CAIR) for patients with esophageal carcinoma were evaluated and compared to conventional irradiation (CR). METHODS: Sixty patients with squamous cell carcinoma of the esophagus were randomized into two groups: the CAIR group (30 patients) and the CR group (30 patients). Patients in the CAIR group received radiotherapy (RT) with 2 Gy/fraction per day at 7 d/wk with a total dose of 50-70 Gy (average dose 64.2 Gy). The overall time of irradiation was 3.6-5.0 wk (average 4.6 wk). RT in the CR group was 2 Gy/fraction per day at 5 d/wk with a total dose of 40-70 Gy (average dose 61.7 Gy). The overall time of irradiation was 4.0-7.0 wk (average 6.4 wk). RESULTS: The data showed that the immediate tumor response to RT was better in the CAIR group than in the CR group. Efficiency rates (CR plus PR) were 82.8% (24/29) and 58.6% (17/29), respectively (P = 0.047). In both groups the incidences of esophagitis and tracheitis were insignificant (P = 0.376, 0.959), and no patient received toxicity that could not be tolerated. CONCLUSION: CAIR shortens overall treatment time and is well tolerated by patients. It may be superior to CR in enhancing the local response of tumor, but its remote effect for esophageal carcinoma awaits further follow up. PMID- 17109505 TI - Application of laparoscopy in diagnosis and treatment of massive small intestinal bleeding: report of 22 cases. AB - AIM: To investigate the diagnostic and therapeutic value of laparoscopy in patients with massive small intestinal bleeding. METHODS: Twenty-two patients with massive small intestinal bleeding and hemodynamic alteration underwent laparoscopic laparotomy in our unit from December 2002 to April 2005. Post pathologic sites were found, laparoscopy- or laparoscopy-assisted part small intestinal resection including pathologic intestinal site and enteroanastomosis was performed in all these patients. RESULTS: The bleeding sites were successfully detected by laparoscopy in all these 22 patients. Massive small intestinal bleeding was caused by jejunum benign stromal tumor in 8 cases, by jejunum potential malignant stromal tumor in 5 cases, by jejunum malignant stromal tumor in 1 case, by Mechel's diverticulum in 5 cases, by small intestinal vascular deformity in 2 cases, and by ectopic pancreas in 1 case. A total of 16 patients underwent laparoscopy-assisted enterectomy and enteroanastomosis of small intestine covering the diseased segment and 6 patients received enterectomy of the diseased segment under laparoscope. No surgical complications occurred and the outcome was satisfactory. CONCLUSION: Laparoscopy in diagnosis and treatment of massive small intestinal bleeding is noninvasive with less pain, short recovery time and definite therapeutic efficacy. PMID- 17109506 TI - Pravastatin: a potential cause for acute pancreatitis. AB - Acute pancreatitis (AP) secondary to drugs is un-common, with an incidence ranging from 0.3% to 2.0% of AP cases. Drug-induced AP due to statins is rare, and only 12 cases have thus far been reported. In this case report, we report a case of a 50-year-old female on pravastatin therapy for 3 d prior to developing symptoms of AP. The common etiological factors for AP were all excluded. The patient was admitted to the intensive care unit secondary to respiratory distress, though she subsequently improved and was discharged 14 d after admission. Although the incidence of drug-induced AP is low, clinicians should have a high index of suspicion for it in patients with AP due to an unknown etiology. Clinicians should be aware of the association of statins with AP. If a patient taking a statin develops abdominal pain, clinicians should consider the diagnosis of AP and conduct the appropriate laboratory and diagnostic evaluation if indicated. PMID- 17109507 TI - Carcinoid of the ampulla of Vater: morphologic features and clinical implications. AB - Carcinoids involving the ampulla of Vater are rare lesions that may produce painless jaundice. The published data indicate that these tumors, in contrast to their midgut counterparts, metastasize in approximately half of cases irrespective of primary tumor size. Therefore, radical excision in the form of pancreaticoduodenectomy is recommended regardless of tumor size. As with other gastrointestinal carcinoid tumors, biological treatment with octreotide analogues can be applied to symptomatic patients. Tumor-targeted radioactive therapy is a newly emerging treatment option. We here report case of a carcinoid tumor of the ampulla of Vater presenting as painless jaundice in a 65-year old man and review the relevant literature, giving special attention to the morphologic features, clinical characteristics, and treatment modalities associated with this disease process. PMID- 17109508 TI - Retroperitoneal fibrosis: a rare cause of both ureteral and small bowel obstruction. AB - Retroperitoneal fibrosis (RPF) is a rare condition of unclear etiology. It can cause ureteral obstruction. We present the unique case of a 54 years old female, who initially presented with spontaneous perforation of the cecum. Upon exploring the abdomen, the classical glistening white, unyielding retroperitoneal fibrosis was encountered. A right hemicolectomy was performed. Subsequently, the patient presented with bilateral ureteral obstruction, and later on with small bowel obstruction. Ureteral obstruction was treated with stents, and small bowel obstruction was treated with bypass. To our knowledge no case of idiopathic RPF presenting with features of both bilateral ureteral and small bowel obstruction has been reported in the literature. PMID- 17109509 TI - How to establish a first-class international scientific journal in China? AB - Hundreds of scientific journals are published in China. However, only scores of them are included in Science Citation Index by the Institute for Scientific Information, with impact factors of only 1 or less. Thus, how to establish a first-class international scientific journal in China is an important but difficult topic that deserves extensive exploration. World Journal of Gastroenterology (WJG) sets a good example although it has experienced setbacks on the road towards success. Concepts and pursuits that affirm the overall development direction, innovation and dreams that provide impetus and aspiration for higher objectives, team work and unique pattern that assure excellent quality and service, and culture and environment that also determine the speed and direction of the development, are believed to be the major factors contributing to the success of WJG. It is recommended that the effective resolution to the above issue is to learn from Chinese examples such as WJG rather than from "how foreign journals do". PMID- 17109510 TI - Current concepts and controversies in the treatment of alcoholic hepatitis. AB - The treatment of alcoholic hepatitis remains one of the most debated topics in medicine and a field of continued research. In this review, we discuss the evolution of scoring systems, including the recent development of the Glasgow alcoholic hepatitis score, role of liver biopsy and current treatment interventions. Studies of treatment interventions with glucocorticoids, pentoxifylline, infliximab, s-adenosyl-methionine, and colchicine are reviewed with discussion on quality. Glucocorticoids currently remain the mainstay of treatment for severe alcoholic hepatitis. PMID- 17109511 TI - Occult hepatitis C virus infection: a new form of hepatitis C. AB - Occult hepatitis C virus (HCV) infection is a new recently characterized entity. This occult infection can be present in two different clinical situations: in anti-HCV negative, serum HCV-RNA negative patients with abnormal liver function tests and in anti-HCV positive subjects with normal values of liver enzymes and without serum HCV-RNA. This review describes recent studies of occult HCV infection in both kinds of patients. PMID- 17109512 TI - Role of ethanol in the regulation of hepatic stellate cell function. AB - Evidence has accumulated to suggest an important role of ethanol and/or its metabolites in the pathogenesis of alcohol-related liver disease. In this review, the fibrogenic effects of ethanol and its metabolites on hepatic stellate cells (HSCs) are discussed. In brief, ethanol interferes with retinoid metabolism and its signaling, induces the release of fibrogenic cytokines such as transforming growth factor beta-1 (TGFbeta-1) from HSCs, up-regulates the gene expression of collagen I and enhances type I collagen protein production by HSCs. Ethanol further perpetuates an activated HSC phenotype through extracellular matrix remodeling. The underlying pathophysiologic mechanisms by which ethanol exerts these pro-fibrogenic effects on HSCs are reviewed. PMID- 17109513 TI - Methylation in esophageal carcinogenesis. AB - Genetic abnormalities of proto-oncogenes and tumor suppressor genes have been demonstrated to be changes that are frequently involved in esophageal cancer pathogenesis. However, hypermethylation of CpG islands, an epigenetic event, is coming more and more into focus in carcinogenesis of the esophagus. Recent studies have proved that promoter hypermethylation of tumor suppressor genes is frequently observed in esophageal carcinomas and seems to play an important role in the pathogenesis of this tumor type. In this review, we will discuss current research on genes that are hypermethylated in human esophageal cancer and precancerous lesions of the esophagus. We will also discuss the potential use of hypermethylated genes as targets for detection, prognosis and treatment of esophageal cancer. PMID- 17109514 TI - Cancer gene therapy targeting angiogenesis: an updated review. AB - Since the relationship between angiogenesis and tumor growth was established by Folkman in 1971, scientists have made efforts exploring the possibilities in treating cancer by targeting angiogenesis. Inhibition of angiogenesis growth factors and administration of angiogenesis inhibitors are the basics of anti angiogenesis therapy. Transfer of anti-angiogenesis genes has received attention recently not only because of the advancement of recombinant vectors, but also because of the localized and sustained expression of therapeutic gene product inside the tumor after gene transfer. This review provides the up-to-date information about the strategies and the vectors studied in the field of anti angiogenesis cancer gene therapy. PMID- 17109515 TI - Comparison of gene expression profiles between primary tumor and metastatic lesions in gastric cancer patients using laser microdissection and cDNA microarray. AB - AIM: To study the differential gene expression profiles of target cells in primary gastric cancer and its metastatic lymph nodes using laser microdissection (LMD) in combination with cDNA microarray. METHODS: Normal gastric tissue samples from 30 healthy individuals, 36 cancer tissue samples from primary gastric carcinoma and lymph node metastasis tissue samples from 58 patients during gastric cancer resection were obtained using LMD in combination with cDNA microarray independently. After P27-based amplification, aRNA from 36 of 58 patients (group 1) with lymph node metastasis and metastatic tissue specimens from the remaining 22 patients (group 2) were applied to cDNA microarray. Semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemical assay verified the results of microarray in group 2 and further identified genes differentially expressed in the progression of gastric cancer. RESULTS: The expression of 10 genes was up-regulated while the expression of 15 genes was down-regulated in 22 gastric carcinoma samples compared with that of genes in the normal controls. The results were confirmed at the level of mRNA and protein, and suggested that four genes (OPCML, RNASE1, YES1 and ACK1) could play a key role in the tumorigenesis and metastasis of gastric cancer. The expression pattern of 3 genes (OPCML, RNASE1 and YES1) was similar to tumor suppressor genes. For example, the expression level of these genes was the highest in normal gastric epithelium, which was decreased in primary carcinoma, and further decreased in metastatic lymph nodes. On the contrary, the expression pattern of gene ACK1 was similar to that of oncogene. Four genes were further identified as differentially expressed genes in the majority of the cases in the progression of gastric cancer. CONCLUSION: LMD in combination with cDNA microarray provides a unique support foe the identification of early expression profiles of differential genes and the expression pattern of 3 genes (OPCML, RNASE1 and YES1) associated with the progression of gastric cancer. Further study is needed to reveal the molecular mechanism of lymph node metastasis in patients with gastric cancer. PMID- 17109516 TI - Efficacy and safety of thalidomide in patients with hepatocellular carcinoma. AB - AIM: To evaluate which patients with hepatocellular carcinoma (HCC) are most likely to respond to thalidomide treatment. METHODS: From July 2002 to July 2004, patients with HCC who received thalidomide treatment, were enrolled. We extracted relevant data from the patients' medical records, including history and type of hepatitis, comorbidity, serum alpha-fetoprotein (alpha-FP) level, volumetric changes in tumor, length of survival, and the dose, duration, side effects of thalidomide treatment. The tumor response was evaluated. On the basis of these data, the patients were divided into two groups: those with either partial response or stable disease (PR + SD group) and those with progressive disease (PD group). RESULTS: Two of 42 (5%) patients had a partial tumor response after treatment with thalidomide, 200 mg/d, and 9 (21%) had stable disease. Patients in the PR + SD group all had cirrhosis. Comparing patients with and without cirrhosis, the former were more likely to respond to thalidomide therapy (PR + SD: 100% vs PD: 64.5%, P = 0.041 < 0.05). Thalidomide was significantly more likely to be effective in tumors smaller than 5 cm (PR + SD: 63.6% vs PD: 25.8%, P = 0.034 < 0.05). Compared with patients with progressive disease (PD), patients in the PR + SD group had a higher total dose of thalidomide (13669.4 +/- 8446.0 mg vs 22022.7 +/- 11461.4 mg, P = 0.023 < 0.05) and a longer survival (181.0 +/- 107.1 d vs 304.4 +/- 167.1 d, P = 0.047 < 0.05). Patients with comorbid disease had a significantly greater incidence of adverse reactions than those without (93.8% vs 60.0%, P = 0.021 < 0.05). The average number of adverse reactions in each person with a comorbid condition was twice as high as in those without other diseases (2.2 +/- 1.3 vs 1.1 +/- 1.2; P = 0.022 < 0.05). CONCLUSION: Thalidomide therapy is most likely to be effective in patients with early stage small HCC, especially in those with other underlying diseases. A low dose (200 mg/d) of thalidomide is recommended to continue the treatment long enough to make it more effective. PMID- 17109517 TI - Proposal of a new and simple staging system of colorectal liver metastasis. AB - AIM: To create a new, simple and useful staging system for colorectal liver metastasis analogous to the Tumor Node Metastasis classification system of International Union Against Cancer. METHODS: A retrospective review was undertaken of 81 consecutive patients who underwent partial hepatectomy for colorectal liver metastases (group 1). Clinical and pathological features of both primary and metastatic liver cancers were entered into a multivariate analysis to determine independent variables helpful in accurately predicting long-term prognosis after hepatectomy. Using selected variables, we created a new staging system like TNM classification. The usefulness of the new staging system was examined in a series of 92 patients from another hospital (group 2). RESULTS: Multivariate analysis showed that 81 patients in group 1 had significant multiple hepatic tumors with the largest tumor being more than 5 cm in diameter, resectable extrahepatic distant metastases, and independent prognostic factors for poor survival after hepatectomy. Using these three variables, we created a new staging system to classify patients with colorectal liver metastases. Finally, our new staging system classified the patients both in group 1 and in group 2. CONCLUSION: Our new staging system of colorectal liver metastasis is simple and useful for staging patients. PMID- 17109518 TI - Expression patterns and action analysis of genes associated with drug-induced liver diseases during rat liver regeneration. AB - AIM: To study the action of the genes associated with drug-induced liver diseases at the gene transcriptional level during liver regeneration (LR) in rats. METHODS: The genes associated with drug-induced liver diseases were obtained by collecting the data from databases and literature, and the gene expression changes in the regenerating liver were checked by the Rat Genome 230 2.0 array. RESULTS: The initial and total expression numbers of genes occurring in phases of 0.5-4 h after partial hepatectomy (PH), 4-6 h after PH (G0/G1 transition), 6-66 h after PH (cell proliferation), 66-168 h after PH (cell differentiation and structure-function reconstruction) were 21, 3, 9, 2 and 21, 9, 19, 18, respectively. It is illustrated that the associated genes were mainly triggered at the initial stage of LR and worked at different phases. According to their expression similarity, these genes were classified into 5 types: only up regulated (12 genes), predominantly up-regulated (4 genes), only down-regulated (11 genes), predominantly down-regulated (3 genes), and approximately up-/down regulated (2 genes). The total times of their up- and down-expression were 130 and 79, respectively, demonstrating that expression of most of the genes was increased during LR, while a few decreased. The cell physiological and biochemical activities during LR were staggered according to the time relevance and were diverse and complicated in gene expression patterns. CONCLUSION: Drug metabolic capacity in regenerating liver was enhanced. Thirty-two genes play important roles during liver regeneration in rats. PMID- 17109519 TI - Meta-analysis on inoperable pancreatic cancer: a comparison between gemcitabine based combination therapy and gemcitabine alone. AB - AIM: To compare gemcitabine-based combination therapy and gemcitabine (GEM) alone in patients with advanced pancreatic cancer (APCa) through meta-analysis. METHODS: MEDLINE and EMBASE searches were supplemented by information from trial registers of randomized controlled trials (RCTs) for GEM-based combination therapy and GEM alone for APCa. A quantitative meta-analysis was carried out by two reviewers based on the inclusion criteria from all available RCTs. The meta analysis involved overall survival (OS), objective remission rate (ORR), clinical benefit rate (CBR), time to progress/progress free survival (TTP/PFS) and toxicity. RESULTS: The meta-analysis included 22 RCTs. There was significant improvement in the GEM combination group with regard to the 6-mo survival rate (RD = 0.04, 95% CI 0.01-0.06, P = 0.008), 1-year survival rate (RD = 0.03, 95% CI 0.01-0.05, P = 0.01), ORR (RD = 0.04, 95% CI 0.01-0.07, P = 0.02), CBR (RD = 0.10, 95% CI 0.02-0.17, P = 0.01) and 6-mo TTP/PFS (RD = 0.07, 95% CI 0.04-0.10, P < 0.00001). However, the Grade 3-4 toxicity set by WHO was higher for the GEM combination group for neutropenia (RD = 0.05, 95% CI 0.01-0.10, P = 0.02), thrombocytopenia (RD = 0.05, 95% CI 0.02-0.08, P = 0.002) and vomiting/nausea (RD = 0.03, 95% CI 0.00-0.05, P = 0.02). CONCLUSION: GEM-based combination therapy may improve the overall survival and palliation in optimal patients with APCa as compared with GEM alone. PMID- 17109520 TI - Relationship between antral distension and postprandial symptoms in functional dyspepsia. AB - AIM: To investigate in patients with functional dyspepsia (FD) after an every-day meal whether (1) gastrointestinal (GI) and extra-GI symptoms had any relation with the degree of antral volume, (2) the onset of postprandial symptoms was associated with, and may predict, delayed gastric emptying. METHODS: In 94 symptomatic FD patients, antral volume variations and gastric emptying were assessed with ultrasonography after a 1050 kcal meal. Symptoms were evaluated with a standardized questionnaire. The association of GI and extra-GI symptoms with antral volumes and gastric emptying were estimated with logistic regression analysis. RESULTS: Forty percent of patients did not report any symptoms after a meal. Compared to the healthy controls, the antrum was more distended in patients throughout the entire observation period and 37 (39.4%) patients had delayed gastric emptying. Only postprandial drowsiness was associated with antral volume variations (AOR = 1.42; P < 0.001) and with delayed gastric emptying (AOR = 3.59; P < 0.03). CONCLUSION: In FD patients, GI symptoms are neither associated with antral distension nor with gastric emptying. Drowsiness is associated with antral distension and delayed gastric emptying. The onset of drowsiness is preceded by an increment of antral distension and the duration of the symptom appears to be related to the persistence of antral distension. PMID- 17109521 TI - Milan criteria are useful predictors for favorable outcomes in hepatocellular carcinoma patients undergoing liver transplantation after transarterial chemoembolization. AB - AIM: To evaluate whether the Milan criteria are useful in patients with hepatocellular carcinoma (HCC) who received transarterial chemoembolization (TACE) before liver transplantation (LT). METHODS: Thirty-six HCC patients who fulfilled the Milan criteria after having received TACE and subsequently underwent LT were included (TACE + LT group) in the study. As controls, 21 patients who also met the Milan criteria and underwent LT without prior treatment were selected (LT group). Post-LT clinical outcomes, such as HCC recurrence, survival rate, and histologic features of explanted livers, were compared between the two groups. RESULTS: Baseline characteristics were not different between the two groups. Pre-LT maximal tumor diameter in TACE + LT group was similar to that of LT group (2.0 +/- 0.6 cm vs 2.3 +/- 0.9 cm; P = 0.10). Post-LT histologic findings also revealed similar maximal tumor diameter in the two groups (2.4 +/- 1.4 cm vs 2.3 +/- 0.9 cm; P = 0.70). Explanted livers showed similar incidence of unfavorable pathologic features. The morality within 60 d after transplantation was not different between the two groups (8.3% vs 9.5%; P = 0.99). Post-LT 5-year survival rate (57% vs 74%; P = 0.70) and cumulative recurrence rate (8.3% vs 4.8%; P = 0.90) were not significantly different between the two groups. CONCLUSION: The Milan criteria are still a useful selection criteria showing favorable outcomes in HCC patients receiving TACE before LT. PMID- 17109522 TI - Adventures in sulfur-nitrogen chemistry. AB - This account reviews our efforts over the past 37 years to understand the chemistry of a select group of sulfur-nitrogen compounds including sulfinimines (N-sulfinyl imines) and N-sulfonyloxaziridines. Our early exploration of the thermal properties of sulfenamides, a class of sulfur-nitrogen compounds about which little was known, resulted in a new procedure, the silver-assisted method, for the construction of sulfenimines (N-sulfenyl imines). Selective oxidations of these compounds resulted in the production of N-sulfinyl imines (sulfinimines) and N-sulfonyloxaziridines. N-Sulfonyloxaziridines turned out to be a new class of aprotic neutral oxidizing reagents. Enantiomerically pure examples afford high ee values in the oxidation of enolates to alpha-hydroxy carbonyl compounds and in the oxidation of sulfides and selenides to sulfoxides and selenoxides. Additions of organometallic reagents to enantiomerically pure sulfinimines provide the best and most versatile method for the asymmetric construction of the carbon-nitrogen stereocenters found in many biologically active compounds. Sulfinimine-derived chiral building blocks provide efficient access to many classes of nitrogen heterocycles including aziridines, 2H-azirines, pyrrolidines, and piperidines. PMID- 17109523 TI - Highly activated vinyl hydrogen in a significantly twisted styrene. AB - The novel example of a vinylic hydrogen more reactive than a benzylic hydrogen was found by treatment of a twisted styrene derivative with a strong base followed by D(2)O quenching. In this paper, the full details of the examples of the highly activated vinyl hydrogens in twisted styrene derivatives are described, with a discussion on the correlation between the reactivity of the vinyl hydrogens and the magnitude of the twist. The highly reactive vinyl hydrogens could be rationalized by considering the novel orbital interaction between the pi(*) orbital of the benzene ring and the pi(*) orbital of the vinylic C-H bond in the twisted styrene derivatives. PMID- 17109524 TI - CASPT2//CASSCF study on the photolysis mechanism of 2,3-diazabicyclo[2.1.1]hex-2 ene: alpha C-N versus beta C-C cleavage. AB - A CASPT2//CASSCF study has been carried out to investigate the mechanism of the photolysis of 2,3-diazabicyclo[2.1.1]hex-2-ene under direct irradiation and triplet-sensitized irradiation. By exploring the detailed potential energy surfaces and surface crossing points for several low-lying excited states, we have been able to provide a tentative description on the photophysical and photochemical processes of this compound. According to our calculations, on the S(1) surface one C-N bond is broken first to generate the diazenyl biradicals, which then result in the photoproduct bicyclobutane through a concerted C-N cleavage and C-C coupling process. The intersystem crossing (ISC) from the S(1) state to the triplet state was found to be quite efficient, which could account for two other photoproducts (butadiene and 1,2-diazabicyclo[3.1.0]hex-2-ene) on direct irradiation. On the T(1) surface, the C-C bond cleavage could readily take place to generate the triplet hydrazonyl biradical. One deactivation path of this intermediate is to convert into the corresponding singlet biradical by an ISC process. Then this biradical undergoes a barrierless C-N bond formation to form the photoproduct 1,2-diazabicyclo[3.1.0]hex-2-ene. Another path of the triplet hydrazonyl biradical is to form the intermediate 2-allyl-diazomethane through a C N cleavage. This intermediate will break another C-N bond to produce a terminal carbene species, which is easily converted into butadiene and bicyclobutane by 1,2-hydrogen shift reaction and addition to C=C double bond, respectively. PMID- 17109525 TI - The photochemistry of 4-azidopyridine-1-oxide. AB - Laser flash photolysis of 4-azidopyridine-1-oxide at 266 or 308 nm yields triplet 4-nitrenopyridine-1-oxide as the dominant reactive intermediate species, with k(ISC) of approximately 2 x 10(7) s(-1). No evidence of products arising from the singlet nitrene was observed, indicating a slow rate of cyclization to the benzazirine and didehydroazepine species. The slow rate of cyclization is postulated to be due to the aminoxyl-like electronic configuration of this species, which withdraws spin density from sites for potential cyclization. PMID- 17109526 TI - Properties of a triazolopyridine system as a molecular chemosensor for metal ions, anions, and amino acids. AB - The characteristics as a chemosensor of the compound 3-methyl-6,8-di(2-pyridyl) [1,2,3]triazolo[5',1':6,1]pyrido[2,3-]pyrimidine (1) have been analyzed. Interaction with Cu(2+) produces a quenching of the fluorescence, while interaction with Zn(2+) leads to a quenching of the fluorescence followed by a bathochromic shift. The crystal structure of the Zn(1)(H(2)O)(3)(ClO(4))(2) x H(2)O complex shows the coordination of Zn(2+) through the terpyridine moiety. The octahedral site is completed by three water molecules. Interactions of the Zn(2+) complex with the anions sulfate, nitrate, nitrite, and dihydrogenphosphate in ethanol produce hypsochromic shifts and restoration of the fluorescence whose magnitude depends on the anion involved. The maximum interaction is observed for H(2)PO(4)(-). Interactions of the Zn(2+) complex with the amino acids l-aspartate and l-glutamate have also been explored showing a higher interaction with l aspartate. PMID- 17109527 TI - A new class of intramolecularly hydrogen-bonded dendrons based on a 2 methoxyisophthalamide repeat unit. AB - The synthesis and conformational properties of folded dendrons based on a 2 methoxyisophthalamide (2-OMe-IPA) repeat unit are described. The hydrodynamic properties of dendrons preorganized via the syn-syn conformational preference of 2-methoxyisophthalamide are compared with 2,6-pyridinedicarboxamide (2,6-pydic) analogues. The effect of subtle differences in the nature of the conformational equilibria that exist within the 2-OMe-IPA and 2,6-pydic repeat units on the global structural properties of the corresponding dendrons was explored computationally, by (1)H-DOSY NMR spectroscopy and time-resolved fluorescence anisotropy (TRFA) measurements. Whereas the syn-syn preference of the 2-OMe-IPA branched repeat unit is stabilized entirely by intramolecular hydrogen-bonding interactions, this preference in the 2,6-pydic system is a consequence of both intramolecular hydrogen-bonding and dipole minimization effects. However, nonspecific solvophobic compression is more important in determining hydrodynamic properties than solvent-dependent shifts in the conformational equilibria of the repeat unit for both dendron series. PMID- 17109528 TI - Aqueous phosphoric acid as a mild reagent for deprotection of tert-butyl carbamates, esters, and ethers. AB - Aqueous phosphoric acid (85 wt %) is an effective, environmentally benign reagent for the deprotection of tert-butyl carbamates, tert-butyl esters, and tert-butyl ethers. The reaction conditions are mild and offer good selectivity in the presence of other acid-sensitive groups, including CBZ carbamates, azetidine, benzyl and methyl esters, TBDMS, and methyl phenyl ethers. The mildness of the reaction is further demonstrated in the synthesis of clarithromycin derivative, in which a tert-butyl ester is removed in the presence of cyclic carbamate, lactone, ketal, acetate ester, and epimerizable methyl ketone functionalities. The reaction preserves the stereochemical integrity of the substrates. The reactions are high yielding, and the workup is convenient. PMID- 17109529 TI - Microwave-assisted preparations of amidrazones and amidoximes. AB - In an operationally straightforward and efficient method, amidrazones and amidoximes are prepared in yields of 65-87% from imidoylbenzotriazoles by microwave heating for 5-20 min with the appropriate hydrazine or hydroxylamine. PMID- 17109530 TI - Synthesis of 1,2,4-trioxepanes and 1,2,4-trioxocanes via photooxygenation of homoallylic alcohols. AB - Homoallylic alcohols 4a-d, easily accessible in two steps from cyclopropyl methyl ketone, underwent a highly regioselective reaction with singlet oxygen to yield gamma-hydroxyhydroperoxides 5a-d in 57-72% yield. Acid-catalyzed reaction of 5a-d with acetone, cyclopentanone, and cyclohexanone furnished 1,2,4-trioxepanes 8a-d, 9a-d, and 10a-d in good yields. Homoallylic alcohol 12 also underwent a highly regioselective photooxygenation to yield gamma-hydroxyhydroperoxide 13 in 67% yield, which on reaction with acetone, cyclopentanone, and cyclohexanone, furnished 1,2,4-trioxocanes 16-18 in 41-55% yield. PMID- 17109531 TI - Acid-base equilibria in nonpolar media. Absolute pK(a) scale of bases in tetrahydrofuran. AB - The acidity constants (pKa) of 11 bases (amines, anilines, pyridines, pyrrolidines, and iminophosphoranes) have been determined in tetrahydrofuran by potentiometry, complemented by conductometric measurements. The pK(a) values of the studied bases cover a wide absolute pH range of acidity in tetrahydrofuran, from 7.4 to 21.7. From the pK(a) values obtained, a scale of absolute acidity in tetrahydrofuran has been established, which has allowed calculation of the absolute pKa values of 77 bases from literature relative pK(a) data. PMID- 17109532 TI - Thermosolvatochromism of merocyanine polarity indicators in pure and aqueous solvents: relevance of solvent lipophilicity. AB - The following novel solvatochromic probes were synthesized: 2,6-dibromo-4-[(E)-2 (1-alkylpyridinium-4-yl)ethenyl] phenolate, where the alkyl groups are methyl, n butyl, n-hexyl, and n-octyl, respectively. Solvatochromism of three of these probes (C(1), C(4), and C(8)) was studied in 36 protic and aprotic solvents. A modified linear solvation energy relationship has been applied to the data obtained at 25 degrees C. Correlation of (empirical) polarities with other solvent properties showed more dependence on lipophilicity than on basicity. A similar conclusion has been reached for a series of other solvatochromic indicators. Exceptions are those that carry acidic hydrogens, being biased toward solvent basicity. Thermosolvatochromism has been studied in mixtures of water with methanol, 1-propanol, acetonitrile, and DMSO. Thermosolvatochromic data have been treated according to a model that explicitly considers the presence in bulk solution of three "species": water, organic component, and solvent-water hydrogen bonded aggregate. Solvation by the latter is favored over solvation by either of the two precursor solvents (aqueous DMSO is an exception). Temperature increase resulted in desolvation of the probes, due to concomitant decrease of the structures of the component solvents. The above-mentioned modified solvation equation has been successfully applied to solvatochromism in aqueous methanol and aqueous 1-propanol. PMID- 17109533 TI - Oligoindenopyrenes: a new class of polycyclic aromatics. AB - A new class of polycyclic aromatic hydrocarbons--oligoindenopyrenes--has been synthesized featuring a Pd-catalyzed Suzuki-Heck coupling cascade. The oligoindenopyrenes are robust, highly colored substructures of C(70) and have properties that might prove useful in new organic materials or devices. After excitation, the tetraindenopyrene derivative 3d undergoes efficient deactivation (99%) by internal conversion to the ground state. The small fluorescence quantum yield (0.004) is in accordance with the short (0.6 ns) fluorescence decay time. PMID- 17109534 TI - Nucleophilic reactivities of indoles. AB - The kinetics of the coupling of indole (1a), N-methylindole (1b), 5-methoxyindole (1c), and 5-cyanoindole (1d) with a set of reference benzhydryl cations have been investigated in acetonitrile and/or dichloromethane. The second-order rate constants for the reactions correlate linearly with the electrophilicity parameter E of the benzhydryl cations. This allows the determination of the reactivity parameters, N and s, characterizing the nucleophilicity of 1a-d according to the linear free enthalpy relationship log k(20 degrees C) = s(N + E) (Acc. Chem. Res. 2003, 36, 66). The nucleophilicity parameters thus defined describe nicely the reactions of 1a-d with 4,6-dinitrobenzofuroxan (2), a neutral superelectrophilic heteroaromatic whose electrophilicity (E) has been recently determined. On this ground, the kinetics of the coupling of 2 with a large variety of indole structures have been studied in acetonitrile, leading to a ranking of this family of pi-excessive carbon nucleophiles over a large domain of the nucleophilicity scale N. Importantly, two linear and parallel correlations are obtained on plotting the measured N values versus the pK(a)(H(2)O) values for protonation at C-3 of 5-X-substituted indoles and 5-X-substituted 2 methylindoles, respectively. This splitting reveals that the presence of the 2 methyl group causes steric hindrance to the approach of 2 from the adjacent C-3 position of an indole structure. The N vs pK(a)(H(2)O) correlation for 5-X substituted indoles is used for a rapid determination of the C-3 basicity of indoles whose acidity constants cannot be measured through equilibrium studies in strongly acidic aqueous media. PMID- 17109535 TI - Electroactive C(2) symmetry receptors based on the biphenyl scaffold and tetrathiafulvalene units. AB - The synthesis of a family of biphenyl-tetrathiafulvalene (TTF) derivatives incorporating a binding site has been carried out in good to moderate yields through functionalization of the biphenyl scaffold. X-ray structure of one derivative (compound 3) of the series is provided and shows a dihedral angle of 74 degrees around the central Ar-Ar bond of the biphenyl unit in a cisoid conformation. (1)H NMR and cyclic-voltammetry studies demonstrate the critical importance of the nature of the substitution on the conformational rigidity and on the electrochemical behavior of the resulting biphenyl-TTF assemblies. This feature is underlined by an original electrochemical recognition process upon binding of Pb(2+), correlated to conformational changes occurring upon metal cation complexation. PMID- 17109536 TI - Design, synthesis, and properties of conformationally fixed semiquinone monoradical species. AB - The design of novel, functionalized semiquinone (SQ) ligands which combine structural rigidity and electron-withdrawing, electron-donating, and electroneutral substituents enables investigation of multiple structure-property relationships and building blocks for new materials, including components of sensors, switches, and molecular spintronics. Along these lines, we report the synthesis of several new SQ ligands containing fused heterocyclic ring systems. Using both electron paramagnetic resonance spectroscopy and quantum chemical calculations, we show how spin density is affected by the fused ring system substituents. PMID- 17109537 TI - Tandem Michael addition/amino-nitrile cyclization from 2-formyl-1,4-DHP in the synthesis of novel dihydroindolizine-based compounds. AB - A simple and efficient methodology for the synthesis of a small library of substituted indolizines with different degrees of saturation starting from the racemic 2-formyl-1,4-DHP reagent was described. The large synthetic possibilities of this reagent as well as of its Knoevenagel corresponding 2-dicyanovinyl-1,4 DHP reagent were investigated using four kinds of activated methylenes as nucleophiles. The key step of the sequential reaction was based on the highly diastereoselective tandem Michael addition/intramolecular amino-nitrile cyclization catalyzed by an organic base, which resulted in the formation of 1,7 dihydroindolizines in a diastereoselective manner. The process seems to be a straightforward one and can be extended to numerous active methylenes such as malononitrile, 1,3-diketones, and alkyl acetoacetates. The 1,3-hydrogen shift of partially hydrogenated indolizines was accomplished easily with a base at room temperature, giving rise to the corresponding 7,8-dihydroindolizines in very good yields. Interestingly, when the active methylene bears a leaving group, the latter process could not be accomplished because a rare cis-elimination of phenylsulfinic acid and nitrous acid preceded the hydrogen shift. The resulting 1,7-dihydroindolizines bearing an exo-methylene group at C1 were not isolated in all cases, as they turned rapidly to indolizines as the thermodynamically more stable products. During these investigations, oxidization of 1,7 dihydroindolizines with CuCl(2) resulted in the formation of polysubstituted pyridines. Also, the epimerization of certain 1,7-dihydroindolizines was evidenced in the solution studied by NMR spectroscopy, whereas in the solid state, they existed only in a unique form as shown by X-ray diffraction analysis of a representative structure. Finally, all products reported herein bear a primary amine and a nitrile function crucial for further transformations. These include the introduction of various pharmacophore groups at either NH(2) or CN groups as well as at both groups at the same time to access the more elaborated indolizines fused to N- or N,N-heterocycles. PMID- 17109538 TI - Palladium-catalyzed coupling of allenylphosphonates, phenylallenes, and allenyl esters: remarkable salt effect and routes to novel benzofurans and isocoumarins. AB - Coupling reactions of allenylphosphonates (OCH(2)CMe(2)CH(2)O)P(O)CH=C=CRR' [R, R' = H (1a), R = H, R' = Me (1b), R = R' = Me (1c)] with aryl iodides, iodophenol, and iodobenzoic acid in the presence of palladium(II) acetate are investigated and compared with those of phenylallenes PhCH=C=CR2 [R = H (2a), Me (2b)] and allenyl esters EtO(2)CCH=C=CR(2) [R = H (2c), Me (2d)]. While 1b and 1c couple with different stereochemical outcomes using PhI in the presence of Pd(OAc)(2)/PPh(3)/K(2)CO(3) to give phenyl-substituted 1,3-butadienes, 1a does not undergo coupling but isomerizes to the acetylene (OCH(2)CMe(2)CH(2)O)P(O)CCMe (7). In the reaction of 1c with PhI, use of K(2)CO(3) affords the butadiene (Z) (OCH(2)CMe(2)CH(2)O)P(O)CH=C(Ph)-C(Me)=CH(2) (12); in contrast, the use of Ag(2)CO(3) leads to the allene (OCH(2)CMe(2)CH(2)O)P(O)C(Ph)=C=CMe(2) (20), showing that these bases differ very significantly in their roles. The reaction of 1a with PhI or PhB(OH)2 in (t)he presence of Pd(OAc)2/CsF/DMF leads mainly to (E)-(OCH(2)CMe(2)CH(2)O)P(O)CH=C(Me)Ph (21) and (OCH(2)CMe(2)CH(2)O)P(O)CH2 C(Ph)=CH(2) (22) and is thus a net 1,2-addition of Ph-H. Compound 1b reacts with iodophenol in the presence of Pd(OAc)(2)/PPh(3)/K(2)CO(3) to give a benzofuran that has a structure different from that obtained by using 1c under similar conditions. Treatment of 1a with iodophenol/Pd(OAc)(2)/CsF/DMF also gives a benzofuran whose structure is different from that obtained by using 2a under similar conditions. In the reaction with 2-iodobenzoic acid, 1a and 2c afford one type of isocoumarin, while 1b,c and 2a,b give a second type of isocoumarin. The structures of key compounds are established by X-ray crystallography. Utility of the phosphonate products in the Horner-Wadsworth-Emmons reaction is demonstrated. PMID- 17109539 TI - Novel carboxylated pyrrole- and carbazole-based monomers. Synthesis and electro oxidation features. AB - Carboxylated pyrrole (Pyr, a index)- and carbazole (Cbz, b index)-containing monomers 6-7a/b and 9a/b have been readily synthesized from the monobenzyl ester of L-glutamic acid and triamine 2 using Clauson-Kaas and amide coupling reactions. In contrast to Pyr-containing compounds 6-7a, and 9a, the three Cbz containing monomers 6-7b, and 9b have been found electroactive and were successfully electropolymerized on a Pt electrode resulting in the deposition of corresponding insoluble electroconducting polyCOOH polyCbz-films poly(6-7b) and poly(9b). PMID- 17109540 TI - Stereocontrolled synthesis of functionalized cis-cyclopentapyrazolidines by 1,3 dipolar cycloaddition reactions of azomethine imines. AB - The reaction of alkene-tethered alpha-ketocarboxylic acid derivatives with monosubstituted hydrazines allows highly substituted cis-cyclopentapyrazolidine ring systems to be constructed rapidly. Successful cyclocondensations are realized under thermal reaction conditions; in some cases, protic or Lewis acids accelerate these reactions. alpha-Methoxy-alpha,beta-unsaturated esters are suitable alkene components, as are alkenes having either electron-withdrawing or electron-donating substituents at the terminal alkene carbon. alpha-Ketoesters, alpha-ketoamides, and alpha-ketothioesters can be employed. Various hydrazines substituted with N-acyl, N-carboalkoxy, or N-carbamothioyl protecting groups are tolerated in these transformations. The rate of intramolecular cycloaddition is found to reflect not only the reactivity and equilibrium concentration of the azomethine imine intermediate, but, also in some cases, the rate at which hydrazone stereoisomers interconvert under the reaction conditions. PMID- 17109541 TI - Hetero diels-alder reaction of vinyl allenes and aldehydes. An experimental and computational study. AB - The hetero Diels-Alder reaction of vinyl allenes and aldehydes in the presence of a Lewis acid has been studied both experimentally and theoretically. Differently substituted vinyl allenes and aldehydes were used to obtain information on the structural requirements of the reaction. Theoretical calculations using the density functional theory indicate that the reaction proceeds through a highly asynchronous polar transition state. PMID- 17109542 TI - Dissolving metal reduction of acetylenes: a computational study. AB - The two-electron, two-proton reduction of alkynes to trans-alkenes has been studied computationally using the polarizable continuum regime to model liquid ammonia, the solvent in which such reductions are generally carried out. Two computational approaches have been used. In one, the energies of species (alkyne, radical anion, vinyl radical, vinyl anion, dianion, and alkene) that are implicated as possible participants in the reduction are obtained using high level ab initio single-point computations under the polarizable continuum model (PCM) conditions. In the other approach, the same species are surrounded by ten explicit ammonia molecules before undergoing the same single-point PCM analysis. It has been shown that the two methods provide nearly identical results in terms of relative energies. Other findings include the probable bent nature of the radical anion species in ammonia, the likelihood that the trans stereochemistry of the reduction is determined at the vinyl anion stage, and the elimination of a dianion as a possible species that determines the stereochemical result. Various observations relating the solvent effects of ammonia are made relative to known gas-phase properties of the species studied. PMID- 17109543 TI - Sigmatropic rearrangements as tools for amino acid and peptide modification: application of the allylic sulfur ylide rearrangement to the preparation of neoglycoconjugates and other conjugates. AB - Reaction of S-allyl cysteine derivatives, generated by the selenocysteine ligation, with rhodium carbenoids, stabilized and unstabilized, enables the attachment of diverse functionality onto cysteine residues. The reaction is successfully applied to the introduction of lipid-like residues, a fluorous alkyl chain, and mono- and disaccharides. PMID- 17109544 TI - Chiral NMR discrimination of piperidines and piperazines using (18-crown-6) 2,3,11,12-tetracarboxylic acid. AB - Enantiomeric discrimination is observed in the (1)H and (13)C NMR spectra of piperidines and piperazines in the presence of (-)-(18-crown-6)-2,3,11,12 tetracarboxylic acid. The amines are protonated by the carboxylic acid groups of the crown ether to produce the corresponding ammonium and carboxylate ions. Association of the ammonium ion with the crown ether likely involves two hydrogen bonds with the crown ether oxygen atoms and an ion pair with the carboxylate anion. Methyl, hydroxymethyl, phenyl, carboxyl, pyridyl, and cyclohexyl substituent groups alpha to the nitrogen atom do not inhibit binding of the ammonium ion to the crown ether. The NMR spectra of piperidines with the stereogenic center alpha or beta to the nitrogen atom exhibit substantial enantiomeric discrimination. Dibasic substrates such as the piperizines are likely converted to their diprotonated form in the presence of the crown ether, and both nitrogen atoms appear to associate with the crown ether moiety. PMID- 17109545 TI - Efficient N-arylation and N-alkenylation of the five DNA/RNA nucleobases. AB - A general approach to N-arylation and N-alkenylation of all five DNA/RNA nucleobases at the nitrogen atom normally attached to the sugar moiety in DNA or RNA has been developed. Various protected or masked nucleobases engaged readily in the copper-mediated Chan-Lam-Evans-modified Ullmann condensation with a range of different boronic acids at room temperature and were subsequently converted to the corresponding deprotected or unmasked adducts. Different N(3)-protecting groups were examined in the case of thymine, where the benzoyl group afforded the highest yields. A 4-alkylthio-substituted pyrimidin-2(1H)-one served as both a cytosine and a uracil precursor and was N-arylated and N-alkenylated in high yields. Adenine was efficiently and selectively N-arylated and N-alkenylated at the N(9) position by employing a bis-Boc-protected adenine derivative, while a bis-Boc-protected 2-amino-6-chloropurine served as guanine precursor and could also be selectively N(9)-arylated and N(9)-alkenylated. PMID- 17109546 TI - Modification of both the electrophilic center and substituents on the nonleaving group in pyridinolysis of O-4-nitrophenyl benzoate and thionobenzoates. AB - A kinetic study is reported for reactions of 4-nitrophenyl benzoate (1c) and O-4 nitrophenyl X-substituted thionobenzoates (2a-e) with a series of pyridines in 80 mol % H2O/20 mol % dimethyl sulfoxide (DMSO) at 25.0 +/- 0.1 degrees C. O-4 Nitrophenyl thionobenzoate (2c) is more reactive than its oxygen analogue 1c toward all the pyridines studied. The Bronsted-type plot is linear with beta(nuc)=1.06 for reactions of 1c but curved for the corresponding reactions of 2c with beta(nu)c decreasing from 1.38 to 0.38 as the pyridine basicity increases, indicating that the reaction mechanism is also influenced on changing the electrophilic center from C=O to C=S. The curvature center of the curved Bronsted-type plots (defined as pK(a)(o)) occurs at pKa = 9.3 regardless of the electronic nature of the substituent X in the nonleaving group. The Hammett plot for reactions of 2a-e with 4-aminopyridine is nonlinear, i.e., the substrates having an electron-donating substituent exhibit negative deviations from the Hammett plot. However, the Yukawa-Tsuno plot for the same reactions exhibits good linear correlation, indicating that the negative deviations shown by these substrates arise from stabilization of the ground state through resonance interaction between the electron-donating substituent X and the C=S bond. PMID- 17109547 TI - Suzuki-Miyaura cross-coupling reactions of benzyl halides with potassium aryltrifluoroborates. AB - [reaction: see text] The palladium-catalyzed cross-coupling of potassium aryltrifluoroborates with benzylic halides occurs in good yield with high functional group tolerance. The increased stability of potassium aryltrifluoroborates compared to other boron coupling partners makes this an effective route to functionalized methylene-linked biaryl systems. PMID- 17109548 TI - Density functional theory study of electroreductive hydrocoupling of alpha,beta unsaturated carbonyl compounds. AB - [reaction: see text] The electroreductive hydrocoupling of methyl cinnamate, methyl crotonate, cumarin, and benzalacetone was studied by DFT (B3LYP/6-311++ G**) calculations. The computational outcomes for the transition states in the hydrocoupling of anion radicals generated by a one-electron transfer to the alpha,beta-unsaturated carbonyl compounds well agree with the diastereoselectivities in the experimental results previously reported. PMID- 17109549 TI - 1,3 Geminal interactions as the possible trend setting factors for C-H and C-C bond energies in alkanes. Support from a density functional theory based bond energy decomposition study. AB - A bond energy decomposition analysis has been carried out to rationalize the well established experimental fact that C-C and C-H bond energies decrease with increasing substitution on the carbon. It is shown that this trend is set by steric 1,3 repulsive interactions (geminal repulsion) that increase in the order 1,3 hydrogen-hydrogen < 1,3 hydrogen-carbon < 1,3 carbon-carbon. On the other hand, the radical stabilization energy has little influence on the observed trend for the C-H bond energy in H-CR(1)R(2)R(3) or the C-C bond energy in H(3)C CR(1)R(2)R(3). Thus, it varies in H-CR(1)R(2)R(3) from -7.2 kcal/mol (H-CH3) to 6.5 kcal/mol (H-C(CH3)3) and in H(3)C-CR(1)R(2)R(3) from -19.0 kcal/mol (H(3)C CH(3)) to -16.9 kcal/mol (H(3)C-C(CH(3))(3)). It was further found that the average intrinsic C-H bond energy in H-CR(1)R(2)R(3) of 129.2 kcal/mol is smaller than the average intrinsic C-C bond energy in H(3)C-CR(1)R(2)R(3) of 143.4 kcal/mol. However, after the inclusion of steric effects, the overall C-H bond becomes stronger than the C-C bond. The role of steric 1,3 repulsive interactions as the trend setting factor has most recently been suggested by Gronert (J. Org. Chem. 2006, 71, 1209) based on an empirical fit of alkane atomization energies. PMID- 17109550 TI - Energetics of oxaspirocycle prototypes: 1,7-dioxaspiro[5.5]undecane and 1,7,9 trioxadispiro[5.1.5.3]hexadecane. AB - The relative gas-phase energetics of several low-lying isomers of 1,7 dioxaspiro[5.5]undecane and 1,7,9-trioxadispiro[5.1.5.3]hexadecane have been calculated with second-order Mller-Plesset perturbation theory and basis sets as large as aug-cc-pVQZ. Relative energies in THF, dichloromethane, acetone, and DMSO have been estimated with corrections from polarized continuum model calculations at the B3LYP/6-311+G(d) level. In the most stable conformation of 1,7-dioxaspiro[5.5]undecane, both rings adopt chair conformations, and both oxygens are axially disposed (2A). It is more than 2 kcal mol(-1) more stable than all the other conformers. In agreement with previous work, the "twist-boat" trans isomer (3A) is the most stable isomer of 1,7,9 trioxadispiro[5.1.5.3]hexadecane. However, in contrast to this earlier study, an "all-chair" conformation (3B) is found to be the most stable cis isomer of 1,7,9 trioxadispiro[5.1.5.3]hexadecane (E approximately 0.5 kcal mol(-1) in acetone and DMSO). Gauge-independent atomic orbital computations at the B3LYP/6-311+G(d) level indicate that this is the only cis isomer with (13)C NMR chemical shifts that are qualitatively consistent with the experimental spectra. PMID- 17109551 TI - Rapid approach to 3,5-disubstituted 1,4-benzodiazepines via the photo-fries rearrangement of anilides. AB - Different anilides derived from carboxylic acids and substituted anilines have been submitted to the photochemically induced Fries rearrangement giving the corresponding o-amino phenones under conditions that are compatible with the presence of acid-labile groups (such as N-Boc or TBDMSO) on R1 and R3. These compounds, not easily obtained in other ways, are useful building blocks for the preparation of benzocondensated heterocycles. After coupling with N-Boc amino acids and TFA-mediated deprotection, the products cyclized to the corresponding 3,5-disubstituted 1,4-benzodiazepin-2-ones, privileged structures predominantly active in the central nervous system. The same results were obtained by coupling with N-Cbz-protected alpha-amino acids followed by microwave assisted hydrogenolysis. When the Fries rearrangement was carried out on the anilide derived from N-Boc-Ala-OH and the further coupling done with N-Cbz-(OMe)Asp-OH, the formed benzodiazepines could be inserted in a peptide chain for the preparation of conformationally constrained peptidomimetics. PMID- 17109552 TI - Grignard reagent-mediated conversion of an acyl nitroso-anthracene cycloadduct to a nitrone. AB - An intramolecular hetero-Diels-Alder cycloadduct of an acyl nitroso compound and a 9,10-dimethyl anthracene derivative was prepared as a potential nitroxyl (HNO) donor. This compound did not release HNO under any of the conditions tested. Treatment of this cycloadduct with excess MeMgCl resulted in the formation of a nitrone, whose structure was confirmed by X-ray crystallography. A mechanism where MeMgCl acts as a nucleophile, strong base, and Lewis acid possibly explains the formation of this product. PMID- 17109553 TI - Triangular assembly through charged hydrogen bonds in polar solvent. AB - We have demonstrated a triangular assembly driven by 12 charged hydrogen bonds and pi-pi interactions. This assembly is stable even in polar solvents because of its charged hydrogen-bond nature. Helical arrangement of the complex is confirmed by its crystal structure. A solution structure is assigned to the D(3) isomer corresponding to the solid-state structure by temperature-dependent (1)H NMR experiments. PMID- 17109554 TI - Synthesis and biological evaluation of azido- and aziridino-hydroxyl-beta-lactams through stereo- and regioselective epoxide ring opening. AB - Two new classes of azido- and aziridino-hydroxyl-beta-lactam containing structures have been prepared by means of a stereo- and regioselective epoxide ring opening. The straightforwardness of the procedure makes this strategy useful for the synthesis of potentially bioactive compounds. Some selected examples showed promising activity in acyl CoA-cholesterol acyltransferase inhibition assays. PMID- 17109555 TI - Efficient synthesis and host-guest properties of a new class of calix[6]azacryptands. AB - Two members of a new class of calix[6]azacryptands, namely, calix[6]tampo and calix[6]tamb, have been synthesized through an efficient [1 + 1] macrocyclization reaction--reduction sequence. One of them has been obtained in a remarkably high overall yield from the known X(6)H(3)Me(3). In comparison to all the other calix[6]azacryptands, they possess unique conformational properties since they present a rigidified cone conformation with a partial filling of the cavity by the methoxy groups. In contrast to calix[6]tampo, the fully protonated derivative of calix[6]tamb behaves as a remarkable molecular receptor toward polar neutral guests. NMR studies have shown that the intracavity binding process is governed by a conformational flip of the aromatic walls of the calixarene core. PMID- 17109556 TI - Highly regio- and stereoselective asymmetric bromoazidation of chiral alpha,beta unsaturated carboxylic acid derivatives: scope and limitations. AB - Lewis acid catalyzed asymmetric bromoazidation of chiral alpha,beta-unsaturated carboxylic acid derivatives was performed using N-bromosuccinimide (NBS) and trimethylsilyl azide (TMSN3) as the bromine and azide sources. Among the Lewis acids, Yb(OTf)3 was found to be the best catalyst. Regio- and anti-selectivity of 100% and moderate to good diastereoselectivity (up to 89:11) with good yields were obtained when Oppolzer's bornane sultam chiral auxilairy was used. Diastereoselectivity of >95:05 was observed when (2S,5S)-2,5-diphenylpyrrolidine was used as the chiral auxiliary. PMID- 17109557 TI - Cascade approach to substituted 6-aryl-phenanthridines from aromatic aldehydes, anilines, and benzenediazonium-2-carboxylate. AB - Aromatic aldehydes reacted with anilines and benzenediazonium-2-carboxylate to afford 6-aryl-phenanthridines. The reaction furnishes a rapid and direct construction of substituted phenanthridine rings from readily available starting materials via a one-pot cascade process. PMID- 17109558 TI - Evolution of pyrrolidine-type asymmetric organocatalysts by "click" chemistry. AB - Click chemistry has been employed to construct a library of the pyrrolidine-type asymmetric organocatalysts. The clicked organocatalysts were evaluated in asymmetric Michael addition of ketones to nitroolefins, showing good catalytic activity and stereoselectivity (up to 100% yield, syn:anti = 99:1, 96% ee). PMID- 17109559 TI - Regio- and stereoselective synthesis of alkenylphosphines: a rhodium-catalyzed hydrophosphination of alkynes using a silylphosphine. AB - A novel rhodium-catalyzed hydrophosphination of alkynes using a silylphosphine as a phosphino group source is described. A variety of alkynes, both terminal and internal ones with aryl, alkyl, and carboxyl groups, gave the corresponding alkenylphosphines in a highly regioselective and syn-selective manner. Alkenes with an electron-withdrawing group also gave the corresponding adducts in good yields. PMID- 17109564 TI - Anti-tumour necrosis factor-alpha therapy for rheumatoid and other inflammatory arthropathies: update on safety in older patients. AB - Anti-tumour necrosis factor (TNF)-alpha represents a major advance in the treatment of rheumatoid arthritis (RA), ankylosing spondylitis and psoriatic arthritis. It is usually well tolerated, but a potential increase in the incidence of some infections in patients taking anti-TNFalpha agents has been reported. Compared with younger people, elderly patients have more co-morbidities and are likely to be taking more medications. Moreover, the aging process induces an increase in the rate of infections. Nevertheless, in recent studies analysing the databases of etanercept trials, the normalised incidence of adverse events, serious adverse events, medically important infections and deaths was not increased in patients aged >or=65 years. However, these trials included patients who might have been healthier than elderly RA patients in the general population and therefore not truly representative. Conflicting results have been reported in several 'real-life' observational studies. Taken together, the available data are reassuring for carefully selected populations, at least for etanercept, but it is not possible to claim that anti-TNFalpha agents do, or do not, pose a particular risk for the general population of older patients. Additional studies aiming at determining the safety and benefit-risk ratio of anti-TNFalpha agents in elderly patients are needed. In addition, since the benefit-risk ratio of anti-TNFalpha agents might be different in patients aged 65, 75 or >80 years, when possible, subgroup analysis might also be useful. PMID- 17109565 TI - Prescribing antiepileptics for the elderly: differences between guideline recommendations and clinical practice. AB - The incidence of epilepsy in patients aged >60 years is higher than in any other period of life. Yet, until recently, what was known about the treatment of older patients with epilepsy has been inferred from studies in younger patients. A growing body of clinical evidence focused exclusively on the elderly suggests that, while some issues are similar for older and younger adults, older patients with epilepsy may require even more attention regarding antiepileptic drug (AED) selection than younger patients. This article reviews published guidelines and recommendations to identify explicit recommendations for use of specific AEDs in the elderly, and assesses the extent to which those recommendations have been adopted in clinical practice. We found that while one systematically derived guideline stated that lamotrigine may be a good choice for older patients because of its favourable adverse effect profile, only clinical recommendations based on expert opinion explicitly identified AEDs that are more and less appropriate for use in the elderly. Examination of published studies describing recent AED prescribing patterns suggests that clinical recommendations have been, at best, slowly adopted. This observation is exemplified by the fact that older patients newly diagnosed with epilepsy are still prescribed phenobarbital--a drug identified as suboptimal in 1985. In order to better understand the delay in adopting clinical recommendations, we examine these findings in light of diffusion of innovations theory, a theory that has been used to understand dissemination of other new medical technologies. According to this theory, while it is too early to suggest that use of second-generation AEDs in the elderly has been delayed, the continued use of phenobarbital in older patients newly diagnosed with epilepsy represents a serious delay in adoption of recent guidelines. Delays may be related to lack of knowledge by primary care clinicians and emergency room physicians (who frequently treat older patients with epilepsy), lack of 'opinion leaders' in primary care and perhaps general neurology, clinicians' focus on seizure control as the primary endpoint in treating patients with epilepsy, and difficulties in changing long-standing prescribing patterns. Research targeting barriers to more appropriate prescribing is needed to determine appropriate strategies for changing AED prescribing practices in the elderly. PMID- 17109566 TI - Divalproex sodium for the treatment of behavioural problems associated with dementia in the elderly. AB - Behavioural and psychological symptoms of dementia (BPSD) are a frequent problem in patients with dementia. The consequences of these neuropsychiatric problems can be significant for both patient and caregiver. Although there is no universally accepted pharmaceutical intervention for the treatment of BPSD, various agents have been studied, including divalproex sodium (valproate semisodium). The results of four placebo-controlled clinical trials are reviewed. None of the studies was sufficient to define clinical practice and the results can be seen as conflicting and inconclusive. Three studies suggested possible short-term efficacy, tolerability and safety of divalproex sodium for agitation and some other neuropsychiatric symptoms associated with dementia in elderly patients; in two of these studies, the findings were obtained only on analysis of secondary outcomes. Benefits of divalproex sodium over placebo were not demonstrated in a fourth study. Further research is needed to determine the optimal use of divalproex sodium for the treatment of neuropsychiatric symptoms of dementia and the long-term benefits, if any, of its use in this patient population. PMID- 17109567 TI - Risperidone for the treatment of neuropsychiatric features in dementia. AB - Neuropsychiatric features of dementia are a common and significant burden on patients and their carers. Management usually involves a combination of pharmacological and non-pharmacological approaches. Antipsychotics are the cornerstone of treatment; among the atypical antipsychotics, risperidone is the most studied. Several trials have confirmed the efficacy of risperidone in the treatment of neuropsychiatric features in dementia, especially for agitation and psychosis. Recent concerns over an increased cerebrovascular adverse event profile and overall mortality with use of antipsychotics have emphasised the need for a risk-benefit analysis for individual demented patients being treated for neuropsychiatric features of their disease. PMID- 17109569 TI - Potentially inappropriate medication prescribing for elderly outpatients in Emilia Romagna, Italy: a population-based cohort study. AB - BACKGROUND: In the US, a growing body of epidemiological studies has documented widespread potentially inappropriate medication prescribing among the elderly in outpatient settings. However, only limited information exists in Europe. OBJECTIVE: To evaluate the prevalence of potentially inappropriate medication prescribing among elderly outpatients in Emilia Romagna, Italy and to investigate factors associated with potentially inappropriate medication prescribing in that setting. METHODS: Retrospective cohort study using the Emilia Romagna outpatient prescription claims database from 1 January 2001 to 31 December 2001 linked with information (age, sex and other variables) available from a demographic file of approximately 1 million Emilia Romagna residents aged >or=65 years. The cohort comprised 849 425 elderly patients who had at least one drug prescription during the study period. The prevalence of potentially inappropriate medication prescribing, as defined by the 2002 Beers' criteria, was measured together with predictors associated with potentially inappropriate medication prescribing. RESULTS: A total of 152 641 (18%) elderly Emilia Romagna outpatients had one or more occurrences of potentially inappropriate medication prescribing. Of these, 11.5% received prescriptions for two medications of concern and 1.7% for three or more. Doxazosin (prescribed to 23% of subjects) was the most frequently occurring potentially inappropriate prescribed medication, followed by ketorolac (20.5%), ticlopidine (18.3%) and amiodarone (12.6%). Factors associated with greater likelihood of potentially inappropriate medication prescribing were older age, overall number of drugs prescribed and greater number of chronic conditions. The odds of receiving potentially inappropriate prescribed medications were lower for females, subjects living in more urban areas and subjects with a higher income level. CONCLUSIONS: This study provides strong evidence that potentially inappropriate medication prescribing for elderly outpatients is a substantial problem in Emilia Romagna. Focusing on the prevalence of potentially inappropriate medication prescribing and associated predictors can help in the development of educational programmes targeting outpatient practitioners to influence prescribing behaviour and, therefore, reduce potentially inappropriate medication prescribing. PMID- 17109572 TI - Acupuncture research: time to shift from theoretical to practical questions. PMID- 17109568 TI - HIV and aging: implications for patient management. AB - Between 2001 and 2004, the percentage of all HIV cases in patients aged >or=50 years increased from 17% to 23%. This concerning increase is expected to continue over the next decade. The increasing prevalence of HIV in these patients is a result of increased longevity in patients treated with highly active antiretroviral therapy (HAART) as well as new primary infections in older patients. While older patients may achieve virological suppression at the same rate as younger patients, the immunological benefit of HAART in older patients may be reduced compared with younger patients. In addition, the toxicities associated with HAART may be worse in older HIV patients, particularly those with underlying renal or hepatic insufficiency. All previous studies evaluating the virological and immunological benefits of HAART in older patients have had relatively small sample sizes and none has compared efficacy or rates of toxicity by HAART treatment class. Co-morbidities are more common in older than in younger patients and can impact on the management of HIV in these patients. Providers must be cognisant of drug-drug interactions and potential adverse effects of HAART regimens when selecting an ideal antiretroviral regimen for older HIV patients. Given the increased longevity and rates of malignancies in HIV-infected patients, providers should also be particularly vigilant in maintaining routine health screening in older HIV patients. Controlled trials on HIV epidemiology, pathogenesis, and therapeutic and clinical outcomes are also needed in older patients. As the HIV-infected population ages, there is a growing need to better determine the effectiveness of HAART in older patients, and to investigate factors associated with a more rapid course of HIV infection in patients aged >50 years. PMID- 17109573 TI - Qigong: life energy and a new science of life. PMID- 17109574 TI - Methodological challenges in whole systems research. PMID- 17109575 TI - A pilot study of external qigong therapy for patients with fibromyalgia. AB - OBJECTIVES: Although qigong is an important part of Traditional Chinese medicine (TCM) based on a philosophy similar to acupuncture, few studies of qigong exist in the Western medicine literature. To evaluate qigong therapy as a modality in treating chronic pain conditions such as fibromyalgia syndrome (FMS), we report a pilot trial of 10 women with severe FMS who experienced significant improvement after external qigong therapy (EQT). DESIGN: Ten patients with FMS completed five to seven sessions of EQT over 3 weeks with pre- and posttreatment assessment and a 3-month follow-up. Each treatment lasted approximately 40 minutes. OUTCOME MEASURES: Tender point count (TPC) and Fibromyalgia Impact Questionnaire (FIQ) were the primary measures. McGill Pain Questionnaire (MPQ), Beck Depression Inventory (BDI), anxiety, and self-efficacy were the secondary outcomes. RESULTS: Subjects demonstrated improvement in functioning, pain, and other symptoms. The mean TPC was reduced from 136.6 to 59.5 after EQT treatment; mean MPQ decreased from 27.0 to 7.2; mean FIQ from 70.1 to 37.3; and mean BDI from 24.3 to 8.3 (all p < 0.01). Many subjects reported reductions in other FMS symptoms, and two reported they were completely symptom-free. Results from the 3-month follow-up indicated some slight rebound from the post-treatment measures, but still much better than those observed at baseline. CONCLUSIONS: Treatment with EQT resulting in complete recovery for some FMS patients suggests that TCM may be very effective for treating pain and the multiplicity of symptoms associated with FMS. Larger controlled trials of this promising intervention are urgently needed. PMID- 17109576 TI - The use of D-ribose in chronic fatigue syndrome and fibromyalgia: a pilot study. AB - OBJECTIVES: Fibromyalgia (FMS) and chronic fatigue syndrome (CFS) are debilitating syndromes that are often associated with impaired cellular energy metabolism. As D-ribose has been shown to increase cellular energy synthesis in heart and skeletal muscle, this open-label uncontrolled pilot study was done to evaluate if D-ribose could improve symptoms in fibromyalgia and/or chronic fatigue syndrome patients. DESIGN: Forty-one (41) patients with a diagnosis of FMS and/or CFS were given D-ribose, a naturally occurring pentose carbohydrate, at a dose of 5 g t.i.d. for a total of 280 g. All patients completed questionnaires containing discrete visual analog scales and a global assessment pre- and post-D-ribose administration. RESULTS: D-ribose, which was well tolerated, resulted in a significant improvement in all five visual analog scale (VAS) categories: energy; sleep; mental clarity; pain intensity; and well-being, as well as an improvement in patients' global assessment. Approximately 66% of patients experienced significant improvement while on D-ribose, with an average increase in energy on the VAS of 45% and an average improvement in overall well being of 30% (p < 0.0001). CONCLUSIONS: D-ribose significantly reduced clinical symptoms in patients suffering from fibromyalgia and chronic fatigue syndrome. PMID- 17109577 TI - Variability in the diagnosis and point selection for persons with frequent headache by traditional Chinese medicine acupuncturists. AB - OBJECTIVES: The aim of this study was to compare Traditional Chinese Medicine (TCM) pattern diagnosis and acupuncture point selection for persons with frequent headache, as ascribed by three highly trained, licensed acupuncturists. METHODS: Thirty-seven (37) study participants with frequent headaches were independently evaluated by three licensed acupuncturists trained in TCM. The acupuncturists identified the meridians and type of dysfunction they believed were contributing to study participants' symptoms. Study acupuncturists also ascribed one or more TCM diagnoses to each participant and selected eight acupuncture points for needling. RESULTS: Some variation in TCM pattern diagnosis and point selection was observed for all subjects. Liver Yang and Qi dysfunction were diagnosed in more than two thirds of subjects. Acupuncture points Liver 3, Large Intestine 4, and Governing Vessel (DU) 20 were the most commonly selected points for treatment. CONCLUSIONS: Headache is a heterogeneous condition represented by a wide variety of TCM diagnoses. There is variability among acupuncturists in the diagnosis of TCM patterns and the selection of acupuncture points for needling. These data suggest, however, that most persons with frequent headache appear to have liver Yang and Qi disharmonies for which needling of Liver 3, Large Intestine 4, and/or Governing Vessel 20 may be appropriate. Further research is needed to determine the extent to which variability in the diagnosis or acupuncture point selection among acupuncturists affects clinical outcomes. PMID- 17109578 TI - Beyond needling--therapeutic processes in acupuncture care: a qualitative study nested within a low-back pain trial. AB - BACKGROUND: In the medical and scientific literature, there is a dearth of reports about how acupuncturists work and deliver care in practice. An informed characterization of the treatment process is needed to support the appropriate design of evaluative studies in acupuncture. METHODS: The design was that of a nested qualitative study within a pragmatic clinical trial. Six acupuncturists who treated up to 25 patients each were interviewed after the treatment phase of the trial to obtain an account of their experiences of providing acupuncture care to patients with low back pain referred by their GP. Using semistructured interviews and a topic guide, data were collected and analyzed for both a priori and emergent themes. This paper focuses on practitioners' accounts of the goals and processes of care, and describes the strategies employed in addition to needling and other hands-on treatments. RESULTS: From the interview data, it is clear that a coherent body of theoretical knowledge informed clinical decisions and practice, and that the goals of treatment went beyond the alleviation of immediate pain-related symptoms. Acupuncturists in this study all described a pattern of patient-centered care based on a therapeutic partnership. Study participants confirmed the importance of three processes that characterized acupuncture care in this trial, each contributing to the goal of a positive long term outcome; building a therapeutic relationship; individualizing care; and facilitating the active engagement of patients in their own recovery. Acupuncturists described elements of care that characterized these processes including establishing rapport, facilitating communication throughout the period of care, using an interactive diagnostic process, matching treatment to the individual patient, and the use of explanatory models from Chinese medicine to aid the development of a shared understanding of the patient's condition and to motivate lifestyle changes that reinforce the potential for a recovery of health. Acupuncturists did not view these therapeutic goals, processes, and strategies as a departure from their usual practice. CONCLUSIONS: This study suggests that acupuncture care for patients with chronic conditions such as low back pain is likely to be a complex intervention that utilizes a number of patient-centered strategies to elicit longterm therapeutic benefits. Research designed to evaluate the effectiveness of acupuncture as it is practiced in the UK needs to accommodate the full range of therapeutic goals and related treatment processes. PMID- 17109579 TI - A single-center, double-blinded, randomized controlled study to evaluate the relative efficacy of sublingual and oral vitamin B-complex administration in reducing total serum homocysteine levels. AB - OBJECTIVE: Reports correlating total homocysteine (tHcy) concentrations with arteriosclerosis have become a matter of interest amongst healthcare professionals and the public. Several commercial preparations of vitamin B complexes have been marketed as supplements intended to reduce elevated levels of tHcy. Among these preparations are those that have been specifically designed for sublingual administration. This study is designed to evaluate the relative efficacy of sublingually versus orally delivered vitamin B complex in reducing serum tHcy levels. DESIGN: Forty-one (41) subjects, between the ages of 50 and 80 years with total serum tHcy concentrations exceeding 11 micromol/L, were treated with a six-week regimen of vitamin B complex. Each B complex consisted of 1000 microg vitamin B12 (as methylcobalamin), 400 microg folate (as folic acid), and 5 mg vitamin B6 (as pyridoxine HCl). Participants in the study were randomized into two groups designated, retrospectively, as SL and PO. Members of group SL were given a sublingually delivered vitamin B complex and a matching orally delivered placebo. Members of group PO were given an orally delivered vitamin complex and a matching sublingually delivered placebo. A statistically significant reduction in tHcy values was observed in both groups upon completion of the 6-week protocol. RESULTS: There was no statistically significant difference in serum tHcy concentrations between SL and PO groups either before or after treatment, substantiating the idea that there is no difference in efficacy between the two methods of vitamin complex delivery. PMID- 17109580 TI - Consumption of Lactobacillus acidophilus and Bifidobacterium longum does not alter phytoestrogen metabolism and plasma hormones in men: a pilot study. AB - OBJECTIVE: The aim of this study was to determine whether equol excretion status and plasma hormone and leptin concentrations can be influenced by consumption of a probiotic supplement. A secondary focus was to investigate whether male equol excretors have a hormone profile consistent with reduced prostate cancer risk. DESIGN: The design was a randomized, single-blinded, placebo-controlled, parallel arm trial. SUBJECTS: Thirty-one (31) of the initially enrolled 39 subjects, 18 to 37 years old, completed all study requirements. INTERVENTION: Subjects consumed either probiotic capsules (containing Lactobacillus acidophilus and Bifidobacterium longum) or placebo capsules for 2 months. Fasting plasma concentrations of testosterone (T), dihydrotestosterone (DHT), androstanediol glucuronide (AAG), androstenedione (A), dehydroepiandrosterone sulfate (DHEAS), sex hormone-binding globulin (SHBG), and leptin were measured on days 1 and 57. Urinary excretion of genistein, glycitein, daidzein, O-desmethylangolensin (O Dma), and equol was measured on days 4 and 61 following a 4-day soy challenge. RESULTS: Probiotic consumption did not significantly alter equol excretor status, plasma hormone, or leptin concentrations in these subjects. At baseline, there were no differences in plasma hormone concentrations between equol excretors and nonexcretors; however, the low number of equol excretors included in this study limits the strength of this finding. CONCLUSIONS: The 2-month intervention with probiotic capsules did not significantly alter equol excretion, plasma hormone, or leptin concentrations in these subjects. A secondary finding was that male equol excretors in this study did not exhibit a hormone profile consistent with reduced prostate cancer risk, although this result should be interpreted with caution. PMID- 17109581 TI - National patterns and correlates of complementary and alternative medicine use in adults with diabetes. AB - OBJECTIVE: The aim of this study was to determine national patterns and correlates of complementary and alternative medicine (CAM) use among adults with diabetes. METHODS: The authors compared CAM use in 2474 adults with and 28,625 adults without diabetes who participated in the most comprehensive national survey on CAM use (2002 National Health Interview Survey). Eight CAM use categories were created, including dietary, herbal, chiropractic, yoga, relaxation, vitamin, prayer, and other (acupuncture, Ayurveda, biofeedback, chelation, energy healing or Reiki therapy, hypnosis, massage, naturopathy, and homeopathy). An overall CAM use category also was created that excluded vitamins and prayer. Patterns of use were compared with chi-square and independent correlates of CAM use with multiple logistic regression controlling for relevant covariates. STATA was used for analysis to account for the complex survey design. RESULTS: Prevalence of overall use of CAM did not differ significantly by diabetes status (47.6 versus 47.9%, p = 0.81). Diabetes was not an independent predictor of overall use of CAM (OR 0.93, 95% confidence interval [CI] 0.83, 1.05). However, persons with diabetes were more likely to use prayer (OR 1.19, 95% CI 1.05, 1.36), but less likely to use herbs (OR 0.86, 95% CI 0.75, 0.99), yoga (OR 0.56, 95% CI 0.43, 0.72), or vitamins (OR 0.82, 95% CI 0.72, 0.93) than people without diabetes after controlling for relevant covariates. Independent correlates of overall use of CAM differed by age, income, employment, comorbidity, and health status between people with and without diabetes. CONCLUSIONS: This study found that there has been a dramatic increase in overall use of CAM in adults with diabetes; diabetes was not an independent predictor of overall use of CAM; and people with diabetes were more likely to use prayer, but less likely to use herbs, yoga, or vitamins compared to persons without diabetes. PMID- 17109582 TI - CAM use among older adults age 65 or older with hypertension in the United States: general use and disease treatment. AB - OBJECTIVES: Hypertension (HT) is a common condition among older adults that greatly increases morbidity and mortality risk. Although a number of antihypertensive therapies are currently available, adherence and control are low. The purpose of this study was to assess the use of complementary and alternative medicine (CAM) among older adults with HT. DESIGN: Using a nationally representative cross-sectional survey, the authors examined the prevalence of CAM use among older adults with and without diagnosed HT and the degree to which CAM is used specifically for HT treatment. SUBJECTS: Subjects were 5821 adults age > or = 65 who participated in the 2002 National Health Interview Survey (NHIS), including the Alternative Health supplement. OUTCOME MEASURES: HT status was assessed by self-report. CAM use was classified as any CAM use, use of four CAM modalities, and specific CAM therapies. RESULTS: Any CAM use was higher for persons with HT compared to those without diagnosed HT (69.5% versus 65.6%). Only 7.8% of CAM users reported using CAM to treat HT. Homeopathic treatment (16.7%), healing ritual (14.5%), and diet-based therapies (13.0%) were the most common HT therapies. Frequency of use of specific types of CAM therapies did not differ by HT status other than for biologically based therapies, which were used less often by those with HT. CONCLUSIONS: CAM use is high among older adults with HT, but the vast majority of CAM is used for treating or preventing other conditions. Healthcare providers treating patients with HT should be aware of CAM among their patients. PMID- 17109583 TI - Using Reiki to decrease memory and behavior problems in mild cognitive impairment and mild Alzheimer's disease. AB - OBJECTIVES: This empirical study explored the efficacy of using Reiki treatment to improve memory and behavior deficiencies in patients with mild cognitive impairment or mild Alzheimer's disease. Reiki is an ancient hands-on healing technique reputedly developed in Tibet 2500 years ago. DESIGN: This study was a quasi-experimental study comparing pre- and post-test scores of the Annotated Mini-Mental State Examination (AMMSE) and Revised Memory and Behavior Problems Checklist (RMBPC) after four weekly treatments of Reiki to a control group. SETTINGS/LOCATION: The participants were treated at a facility provided by the Pleasant Point Health Center on the Passamaquoddy Indian Reservation. SUBJECTS: The sample included 24 participants scoring between 20 and 24 on the AMMSE. Demographic characteristics of the sample included an age range from 60 to 80, with 67% female, 46% American Indian, and the remainder white. INTERVENTIONS: Twelve participants were exposed to 4 weeks of weekly treatments of Reiki from two Reiki Master-level practitioners; 12 participants served as controls and received no treatment. OUTCOME MEASURES: The two groups were compared on pre- and post-treatment scores on the AMMSE and the Revised Memory and Behavior Problems Checklist (RMBPC). RESULTS: Results indicated statistically significant increases in mental functioning (as demonstrated by improved scores of the AMMSE) and memory and behavior problems (as measured by the RMBPC) after Reiki treatment. This research adds to a very sparse database from empirical studies on Reiki results. CONCLUSION: The results indicate that Reiki treatments show promise for improving certain behavior and memory problems in patients with mild cognitive impairment or mild Alzheimer's disease. Caregivers can administer Reiki at little or no cost, resulting in significant societal value by potentially reducing the needs for medication and hospitalization. PMID- 17109584 TI - "Torque-like" action of remedies and diseases on the vital force and their consequences for homeopathic treatment. AB - INTRODUCTION: Within the developing theoretical context of quantum macroentanglement, a mathematical model of the Vital Force (Vf) has recently been formulated. It describes the Vf in terms of a hypothetical gyroscope with quantized angular momentum. This enables the Vf's state of health to be represented in terms of a "wave function" derived solely from secondary symptom observables produced in response to disease or homeopathic remedies. So far, this approach has illustrated the biphasal action of remedies, resonance phenomena arising out of homeopathic provings, and aspects of the therapeutic encounter. METHOD: In this paper, the effects of a therapeutic remedy and disease states on the gyroscopic angular momentum characteristics of the Vf were investigated by mathematical manipulation the Vf "wave function." RESULTS: The formulation of the Vf in terms of a quantized gyroscope permits disease to be represented as a braking "torque" decreasing the Vf's angular momentum, causing it to "precess" (i.e., express symptoms). The therapeutic remedy, however, provides an accelerating "torque," increasing the Vf's angular momentum and decreasing precession (i.e., reducing symptom expression). CONCLUSIONS: According to this model, symptom expression corresponds to precession of the Vf "gyroscope." Conversely, complete removal of symptoms is equivalent to cessation of Vf "precession." However, if overprescribed or given in unsuitable potency, the curative remedy (which may also be formulated as a wave function but this time derived solely from changes in Vf secondary symptom observables) may cause the Vf to express proving symptoms. Thus, with only observation of symptoms and changes in them to indicate, indirectly, the state of a patient's Vf, the safest treatment strategy might be for the practitioner to proceed via gradual removal of the symptoms. This is congruent with Hahnemann's later development and use of the LM potencies, as described in his final 6th edition of The organon. PMID- 17109585 TI - Neurobiological correlates of acupuncture: November 17-18, 2005. AB - The "Neurobiological Correlates of Acupuncture" Conference was convened November 17-18, 2005 in Bethesda, Maryland. The conference was sponsored by the National Center for Complementary and Alternative Medicine (NCCAM) of the National Institutes of Health (NIH), U.S. Department of Health and Human Services (DHHS). Its goals were to encourage exchange of ideas regarding the direction of neuroimaging in acupuncture research as well as to discuss some of the challenges in this field. The use of neuroimaging, a relatively recent advance in the study of acupuncture, holds the promise of localizing and characterizing brain activity associated with acupuncture interventions in real time and in a minimally invasive way. Among the main challenges to research into the biological mechanisms of acupuncture are the multiple treatment variables and the difficulties of selecting appropriate experimental controls. Despite these challenges, numerous findings from acupuncture neuroimaging experiments were presented and discussed at the conference on topics related to possible signaling networks, sham acupuncture controls, acupoint specificity, acupuncture analgesia, acupuncture-associated brain response, and the potential for using neuroimaging in conjunction with translational and clinical acupuncture research. Future directions in acupuncture neuroimaging research, as recommended by conference participants, should focus on (1) continuing exploration of acupuncture signaling networks; (2) establishing standards and recommendations for performing and reporting acupuncture neuroimaging results; (3) enabling data sharing in the acupuncture neuroimaging community; (4) gaining a better understanding of placebo and control groups in acupuncture neuroimaging experiments; and (5) developing biomarkers that relate to physiologically and/or clinically relevant acupuncture responses to neuroimaging results. PMID- 17109592 TI - Comparison of once-daily glargine insulin with twice-daily NPH/regular insulin for control of hyperglycemia in inpatients after cardiovascular surgery. AB - BACKGROUND: Traditionally hyperglycemia in surgical inpatients has been managed with six-hourly sliding-scale regular insulin. However, this approach is usually ineffective in preventing hyperglycemia since no basal insulin is provided. We compared glycemic control using NPH and regular insulin versus glargine insulin alone in patients after cardiovascular surgery on a general surgical ward. METHODS: Ninety-four hyperglycemic patients were randomized to subcutaneous insulin using twice-daily NPH/regular or once-daily glargine if they required at least 1 unit/h of intravenous insulin at the time of transfer from the ICU. NPH/regular was adjusted twice daily; glargine was adjusted once daily. Blood glucose was measured four times daily and targeted to 80-140 mg/dL. RESULTS: The mean blood glucose after NPH/regular (124 mg/dL) and glargine (131 mg/dL) was similar (P = 0.065). In the subgroup of patients with a history of diabetes, mean blood glucose was significantly lower after NPH/regular (133 mg/dL) versus glargine (154 mg/dL) (P = 0.016). Blood glucose less than 60 mg/dL was significantly less common after glargine (0.5%) as compared with NPH/regular (2%) (P = 0.036). CONCLUSIONS: Once-daily glargine insulin provides good glycemic control in hyperglycemic patients after cardiovascular surgery. Although twice daily NPH/regular insulin provided better control than glargine insulin monotherapy, the simplicity and safety of glargine insulin make it an attractive option for the management of postoperative hyperglycemia. Patients with established diabetes will achieve better glucose control with NPH/regular insulin as compared with glargine but have a higher incidence of hypoglycemia. PMID- 17109593 TI - Dynamic modeling of free fatty acid, glucose, and insulin: an extended "minimal model". AB - BACKGROUND: The mathematical models for patients with diabetes proposed in the literature since the late 1970s are mainly glucocentric (glucose-based); hence, the contribution of free fatty acid (FFA) metabolism in the body and its glucose insulin interactions have been largely ignored. However, approximately 90% of the muscle energy is derived from FFA metabolism when the body is at rest. Furthermore, significant interactions exist among FFA, glucose, and insulin. With the long-term goal of developing a closed-loop glucose control system, a model of the major energy-providing substrate dynamics is required. METHODS: The Bergman minimal model was extended to include plasma FFA dynamics, and its interaction with glucose and insulin dynamics, with a primary focus on patients with Type 1 diabetes. Differential equations were developed for plasma FFA concentrations and "remote" FFA effects on glucose uptake, as well as "remote" insulin effects on plasma FFA concentrations. Parameters for the model were estimated from experimental data provided in the scientific literature. RESULTS: The minimal model was extended in order to capture three major metabolic aspects: the antilipolytic effect of insulin; the lipolytic effect of prolonged hyperglycemia; and the impairing effect of FFA on glucose uptake rate. The dynamic fit of glucose, FFA, and insulin profiles is consistent with published data. CONCLUSIONS: The extended minimal model successfully captured the plasma FFA concentration behavior, the plasma insulin and glucose concentrations, and the physiological interactions that exist among these species. This more comprehensive description of energy-providing substrate dynamics may provide a novel simulation test-bed for analysis of patients with insulin dependent diabetes and controller design. PMID- 17109594 TI - Efficacy of continuous real-time blood glucose monitoring during and after prolonged high-intensity cycling exercise: spinning with a continuous glucose monitoring system. AB - BACKGROUND: Hypoglycemia is the most common and serious side effect of insulin therapy in type 1 diabetes (T1DM), frequently occurring both during and after vigorous exercise. Late-onset hypoglycemia (LOH) is of great concern, occurring 1 36 h after exercise, often going unnoticed during sleep. Repeated exposure to LOH causes autonomic glucose counterregulatory failure and sometimes coma and death. Continuous glucose monitoring systems have recently emerged as a potentially important tool in diabetes management, allowing individuals to track glucose levels continuously and learn how various behaviors influence glucose control. METHODS: In this pilot study, we determined the efficacy of using a real-time continuous glucose monitoring system (Guardian RT, Minimed, Northridge, CA) to detect blood glucose excursions associated with exercise and LOH (i.e., blood glucose concentration <4 mM) after exercise in individuals with T1DM. Five subjects with T1DM were monitored before, during, and after a 60 min vigorous spin class using Guardian RT (48 h in total). RESULTS: Following the exercise, three of the five subjects had LOH, while the other two experienced decreases in blood glucose concentrations to 4 mM. The Guardian RT monitor was effective in notifying all of the subjects of such glycemic excursions over the 48 h surveillance period. A strong correlation (r = 0.89, P < 0.001) was found between conventional self-monitoring of blood glucose and Guardian RT data pairs. CONCLUSION: These limited data suggest that nocturnal LOH occurs commonly following vigorous exercise and that a Guardian RT is a useful and important diagnostic tool. Further study into clinical strategies for preventing hypoglycemia associated with this common form of mixed aerobic and anaerobic exercise is urgently needed through insulin modification and carbohydrate supplementation. PMID- 17109595 TI - The effect of chromium picolinate and biotin supplementation on glycemic control in poorly controlled patients with type 2 diabetes mellitus: a placebo controlled, double-blinded, randomized trial. AB - BACKGROUND: Preclinical studies have shown that the combination of chromium picolinate and biotin significantly enhances glucose uptake in skeletal muscle cells and enhances glucose disposal. The present pilot study was conducted to determine if supplementation with chromium picolinate and biotin can improve glycemic control in patients with type 2 diabetes mellitus with suboptimal glycemic control despite use of oral antihyperglycemic agents. METHODS: Forty three subjects with impaired glycemic control (2-h glucose >200 mg/dL; glycated hemoglobin >or=7%), despite treatments with oral antihyperglycemic agents, were randomized to receive 600 microg of chromium as chromium picolinate and biotin (2 mg/day) (Diachrome(, Nutrition 21, Inc., Purchase, NY) in addition to their prestudy oral antihyperglycemic agent therapy. Measurements of glycemic control and blood lipids were taken at baseline and after 4 weeks. RESULTS: After 4 weeks, there was a significantly greater reduction in the total area under the curve for glucose during the 2-h oral glucose tolerance test for the treatment group (mean change -9.7%) compared with the placebo group (mean change +5.1%, P < 0.03). Significantly greater reductions were also seen in fructosamine (P < 0.03), triglycerides (P < 0.02), and triglycerides/ high-density lipoprotein cholesterol ratio (P < 0.05) in the treatment group. No significant adverse events were attributed to chromium picolinate and biotin supplementation. CONCLUSIONS: This pilot study demonstrates that supplementation with a combination of chromium picolinate and biotin in poorly controlled patients with diabetes receiving antidiabetic therapy improved glucose management and several lipid measurements. Chromium picolinate/ biotin supplementation may represent an effective adjunctive nutritional therapy to people with poorly controlled diabetes with the potential for improving lipid metabolism. PMID- 17109596 TI - A novel analytical method for assessing glucose variability: using CGMS in type 1 diabetes mellitus. AB - BACKGROUND: Marked blood glucose (BG) fluctuations may increase the risk of some complications associated with diabetes. Acute BG excursions are common in patients with diabetes, but are not usually quantified, nor can they be captured by glycosylated hemoglobin level. This study evaluated the sensitivity of novel analytical methods for assessing BG variability using CGMS (Medtronic Minimed, Northridge, CA) data from patients treated with pramlintide, a drug that acutely reduces postprandial hyperglycemia when added to insulin therapy. METHODS: Retrospective analyses were done on 24-h CGMS profiles obtained from 22 evaluable subjects with type 1 diabetes using insulin pumps and receiving preprandial three times daily injections of placebo (n = 6) or 30 microg of pramlintide (n = 16) for 4 weeks. CGMS data were recorded at baseline, after 4 weeks of treatment, and after 2 weeks off-treatment. Three parameters were calculated for each time period: variability (BG rate of change), an index for severe hypoglycemia [low BG index (LBGI)], and an index for marked hyperglycemia [high BG index (HBGI)]. RESULTS: The mean postprandial BG rate of change was significantly lower after 4 weeks of pramlintide treatment compared with placebo treatment (0.87 vs. 1.21 mg/dL/min, P < 0.01) without changes in average glycemia, illustrating the sensitivity of this parameter to medication effects. The HBGI and LBGI indicated a decreased risk of hyperglycemia without a significant increase in risk of hypoglycemia after 4 weeks of pramlintide. CONCLUSIONS: These results suggest the potential utility of several novel methods for assessing variability and glycemic extremes to gauge the effects of pharmacological interventions not captured by glycosylated hemoglobin. PMID- 17109597 TI - Diabetic nerve conduction abnormalities in the primary care setting. AB - BACKGROUND: Nerve conduction studies (NCS) are the most objective measure of nerve function, and their use is recommended in the clinical and epidemiological evaluation of diabetic polyneuropathy (DPN). The purpose of this study was to utilize automated NCS technology to characterize nerve conduction of patients with diabetes in primary care settings. METHODS: The Diabetes cohort was drawn from 28 community clinics. The Control cohort consisted of subjects without diabetes and without evidence of neuropathy. Bilateral peroneal NCS were performed with an automated NCS instrument (NC-stat, NeuroMetrix, Inc., Waltham, MA). Neuropathic symptoms were quantified using an abbreviated form of the NTSS-6 questionnaire. Risk factors for abnormal NCS were determined using multivariate regression modeling. RESULTS: Data were collected for 172 control subjects and 1,358 subjects with diabetes. Statistically significant differences in peroneal NCS were found. Of the Diabetes cohort, 75.1% had at least one NCS abnormality, and 53.2% had bilateral abnormalities. Of the asymptomatic patients, 45% had bilateral NCS abnormalities. By contrast, 40% of those with clinically significant symptoms lacked bilateral NCS abnormalities. Independent predictors for bilateral NCS abnormalities were age, height, weight, hemoglobin A1c (HbA1c), and duration of diabetes. Up to 16% of the variance in NCS measurements was explained by HbA1c, duration of diabetes, and several demographic variables. CONCLUSIONS: This study suggests that automated NCS can provide nerve conduction confirmation of DPN in primary care settings and has clinical utility. These findings have important implications for the clinical and epidemiological evaluation of DPN. PMID- 17109598 TI - Insulin pump therapy from the time of diagnosis of type 1 diabetes. AB - BACKGROUND: This study was designed to test the feasibility and efficacy of continuous subcutaneous insulin infusion (CSII) being instituted within 1 month of diagnosis of type 1 diabetes mellitus (T1DM). METHODS: Twenty-eight consecutive patients with newly diagnosed T1DM with a mean age of 12.1 +/- 6.2 years were placed on CSII, as early as within 1 day of their diagnosis. All accepted CSII when offered it, and none elected to discontinue CSII after follow up periods of up to 3 years. RESULTS: Hemoglobin A1c levels declined from an initial mean of 10.5 +/- 2.4% to between 6.5% and 7.4% over the next 18 months, at a mean insulin requirement of 0.33 units/kg/day at 3 months, which gradually rose to 0.58 units/kg/day by 18 months. Endogenous insulin secretion, measured by C-peptide values, remained stable during the first 12 months after diagnosis. There was no significant weight gain for the duration of the study (20.7 kg/m(2) vs. a peak of 22.1 kg/m(2) at 12 months, P = 0.54). CONCLUSIONS: The study provided a positive experience with CSII as the initial insulin replacement therapy in newly diagnosed patients with T1DM with excellent clinical outcomes and apparent prolongations of the honeymoon period. It remains to be proven by random patient assignment whether endogenous insulin secretion is better preserved with CSII as an initial ongoing treatment modality and whether long term complications are reduced by this approach. PMID- 17109599 TI - Bedside monitoring of blood beta-hydroxybutyrate levels in the management of diabetic ketoacidosis in children. AB - INTRODUCTION: Diabetic ketoacidosis (DKA) affects many children with type 1 diabetes. Insulin treatment of DKA is traditionally guided by changes in the blood glucose levels and blood gases, whereas beta-hydroxybutyrate (beta-OHB)- the main ketoacid causing acidosis--is rarely measured. The purpose of this study was to evaluate if bedside monitoring of blood beta-OHB levels can simplify management of DKA through elimination of superfluous laboratory monitoring. METHODS: Our emergency department treated 68 children with DKA using a standard protocol with monitoring of venous pH, partial pressure of CO(2) (pCO(2)), bicarbonate, glucose, blood urea nitrogen, and electrolytes (two to 10 time points per patient). Venous beta-OHB levels were measured using the Precision Xtra meter (MediSense/Abbott Diabetes Care, Abbott Park, IL) and, on duplicate batched serum samples, using a reference laboratory method (Cobas Mira Plus; Roche Diagnostics, Indianapolis, IN). Correlations between bedside meter beta-OHB and other parameters were evaluated in a series of general linear models with a time series covariance structure fit using spatial power law. RESULTS: The bedside meter beta-OHB levels were significantly correlated with pH (r = -0.63; P <0.0001), bicarbonate (r = -0.74; P <0.0001), and pCO(2) (r = -0.55; P <0.0001) at all points of measurement during the treatment (unadjusted Pearson correlations). The pH, bicarbonate, and pCO(2) were entered into separate time series analysis models with treatment duration as a measure of time. The results confirmed that bedside levels of beta-OHB correlated very closely with time dependent levels of venous pH, bicarbonate, and pCO(2). Good agreement between the two methods of beta-OHB measurement (r = 0.92; P <0.0001) was confirmed using the Bland-Altman plot analysis. CONCLUSIONS: The Precision Xtra accurately measures blood beta-OHB levels, particularly at lower levels. While the initial measurement of pH and/or bicarbonates is warranted, real-time beta-OHB levels may replace repeat laboratory measurement of these parameters in the management of DKA. Future studies should evaluate safety and cost-effectiveness of such simplified DKA treatment protocol. PMID- 17109600 TI - Clinical studies on chromium picolinate supplementation in diabetes mellitus--a review. AB - Chromium (Cr) picolinate (CrPic) is a widely used nutritional supplement for optimal insulin function. A relationship among Cr status, diabetes, and associated pathologies has been established. Virtually all trials using CrPic supplementation for subjects with diabetes have demonstrated beneficial effects. Thirteen of 15 clinical studies (including 11 randomized, controlled studies) involving a total of 1,690 subjects (1,505 in CrPic group) reported significant improvement in at least one outcome of glycemic control. All 15 studies showed salutary effects in at least one parameter of diabetes management, including dyslipidemia. Positive outcomes from CrPic supplementation included reduced blood glucose, insulin, cholesterol, and triglyceride levels and reduced requirements for hypoglycemic medication. The greater bioavailability of CrPic compared with other forms of Cr (e.g., niacin-bound Cr or CrCl(3)) may explain its comparatively superior efficacy in glycemic and lipidemic control. The pooled data from studies using CrPic supplementation for type 2 diabetes mellitus subjects show substantial reductions in hyperglycemia and hyperinsulinemia, which equate to a reduced risk for disease complications. Collectively, the data support the safety and therapeutic value of CrPic for the management of cholesterolemia and hyperglycemia in subjects with diabetes. PMID- 17109601 TI - Improving laboratory diagnosis of diabetic nephropathy with the use of glomerular filtration rate. PMID- 17109602 TI - Chloroquine-induced nitric oxide: new treatment for an emerging epidemic of obesity-related glomerulopathy. PMID- 17109604 TI - Linkage identification by fitness difference clustering. AB - Genetic Algorithms perform crossovers effectively when linkage sets - sets of variables tightly linked to form building blocks - are identified. Several methods have been proposed to detect the linkage sets. Perturbation methods (PMs) investigate fitness differences by perturbations of gene values and Estimation of distribution algorithms (EDAs) estimate the distribution of promising strings. In this paper, we propose a novel approach combining both of them, which detects dependencies of variables by estimating the distribution of strings clustered according to fitness differences. The proposed algorithm, called the Dependency Detection for Distribution Derived from fitness Differences (D(5)), can detect dependencies of a class of functions that are difficult for EDAs, and requires less computational cost than PMs. PMID- 17109605 TI - Understanding the biases of generalised recombination: part I. AB - This is the first part of a two-part paper where we propose, model theoretically and study a general notion of recombination for fixed-length strings, where homologous recombination, inversion, gene duplication, gene deletion, diploidy and more are just special cases. The analysis of the model reveals that the notion of schema emerges naturally from the model's equations. In Part I, after describing and characterising the notion of generalised recombination, we derive both microscopic and coarse-grained evolution equations for strings and schemata and illustrate their features with simple examples. Also, we explain the hierarchical nature of the schema evolution equations and show how the theory presented here generalises past work in evolutionary computation. In Part II, the study provides a variety of fixed points for evolution in the case where recombination is used alone, which generalise Geiringer's theorem. In addition, we numerically integrate the infinite-population schema equations for some interesting problems, where selection and recombination are used together to illustrate how these operators interact. Finally, to assess by how much genetic drift can make a system deviate from the infinite-population-model predictions we discuss the results of real GA runs for the same model problems with generalised recombination, selection and finite populations of different sizes. PMID- 17109606 TI - Evolving combinatorial problem instances that are difficult to solve. AB - This paper demonstrates how evolutionary computation can be used to acquire difficult to solve combinatorial problem instances. As a result of this technique, the corresponding algorithms used to solve these instances are stress tested. The technique is applied in three important domains of combinatorial optimisation, binary constraint satisfaction, Boolean satisfiability, and the travelling salesman problem. The problem instances acquired through this technique are more difficult than the ones found in popular benchmarks. In this paper, these evolved instances are analysed with the aim to explain their difficulty in terms of structural properties, thereby exposing the weaknesses of corresponding algorithms. PMID- 17109607 TI - Introducing robustness in multi-objective optimization. AB - In optimization studies including multi-objective optimization, the main focus is placed on finding the global optimum or global Pareto-optimal solutions, representing the best possible objective values. However, in practice, users may not always be interested in finding the so-called global best solutions, particularly when these solutions are quite sensitive to the variable perturbations which cannot be avoided in practice. In such cases, practitioners are interested in finding the robust solutions which are less sensitive to small perturbations in variables. Although robust optimization is dealt with in detail in single-objective evolutionary optimization studies, in this paper, we present two different robust multi-objective optimization procedures, where the emphasis is to find a robust frontier, instead of the global Pareto-optimal frontier in a problem. The first procedure is a straightforward extension of a technique used for single-objective optimization and the second procedure is a more practical approach enabling a user to set the extent of robustness desired in a problem. To demonstrate the differences between global and robust multi-objective optimization principles and the differences between the two robust optimization procedures suggested here, we develop a number of constrained and unconstrained test problems having two and three objectives and show simulation results using an evolutionary multi-objective optimization (EMO) algorithm. Finally, we also apply both robust optimization methodologies to an engineering design problem. PMID- 17109609 TI - Emerging treatments for depression. AB - Established treatments for depression are often effective. However, a significant number of patients show limited or no response. With advancements in the explanation of the underlying neurobiology of depression, several novel therapeutic approaches have been developed. Emerging drug targets include novel monoamine oxidase inhibitors, triple monoamine re-uptake inhibitors, omega-3 fatty acids, melatoninergic agonists and receptor antagonists for corticotropin releasing factor(1), glucocorticoid, substance-P and NMDA. Developments in therapeutic focal brain stimulation include vagus nerve stimulation, transcranial magnetic stimulation, magnetic seizure therapy and deep brain stimulation. The role of psychotherapy, both as monotherapy and as adjunctive therapy, is an active avenue of investigation. Although data on these treatments are limited, preliminary results are encouraging. A major goal that remains to be achieved is the identification of predictors of response to the various antidepressant treatments that have diverse mechanisms of action. PMID- 17109610 TI - New pharmacological approaches for the treatment of alcoholism. AB - Pharmacological relapse prevention in alcoholism is a rather new clinical field with few drugs being available. Acamprosate, acting predominantly via glutamatergic pathways, and the opioid receptor antagonist naltrexone, were both shown to be efficient in improving rates for continuous abstinence, and not relapsing to heavy drinking in a number of clinical trials and meta-analyses. There are conflicting data on both drugs, especially for acamprosate, according to some recent US studies. However, overall, the evidence is good. Both drugs are approved in most European countries and the US. Efficacy data for disulfiram are mixed; it is a second-line medication compared with other drugs, and is probably most effective when used in a supervised setting. Recently, anticonvulsants including carbamazepine and topiramate have been discussed as possible anti craving drugs, but there is still limited evidence for their efficacy. Although there is a significant comorbidity for alcoholism with affective disorder, anxiety and schizophrenia, relatively few controlled clinical trials have been performed in this area. Tricyclics have been found to be more effective than serotonin reuptake inhibitors in improving depressive symptoms in these patients. PMID- 17109611 TI - A review of drug options in age-related macular degeneration therapy and potential new agents. AB - Age-related macular degeneration (AMD) is the leading cause of legal blindness in people > 50 years of age in the developed world. AMD is both a debilitating and costly disease for the individual and the community. Greater understanding of the mechanisms and pathways involved in causing the visual loss in AMD has resulted in the advent of several newer and more effective treatment options, making it an exciting time in the management of AMD. This paper will examine the principles behind the existing drug therapies available, as well as those being developed in the management or prophylaxis of AMD and its vision-threatening complications. PMID- 17109612 TI - Treatment of allograft vasculopathy in heart transplantation. AB - Cardiac allograft vasculopathy remains one of the main causes of morbidity and mortality after heart transplantation, although its impact is becoming somewhat smaller as prophylactic measures are implemented. Advances in the understanding of the molecular and cellular mechanisms involved in the genesis and development of cardiac allograft vasculopathy are opening ways for new diagnostic and therapeutic strategies. Successful prophylaxis of the early stages of the disease has been demonstrated with the use of newer immunosuppressive agents, such as sirolimus and everolimus, that will probably be included in future protocols. For most patients with established cardiac allograft vasculopathy, currently available revascularisation methods and retransplantation are not appropriate options. Antiproliferative agents could provide significant improvement in terms of symptom relief and prognosis, but their definite value must be proven in well designed trials. PMID- 17109613 TI - Calcium channel blockers in the spectrum of antihypertensive agents. AB - Calcium channel blockers (CCBs) are widely used in the treatment of hypertension. Through blood pressure reduction, and possibly other mechanisms such as antioxidative effects, they may play a role in diminishing the risk for a variety of cardiovascular outcomes. The combination of CCBs with other newer antihypertensive agents such, as ACE inhibitors and angiotensin receptor blockers, may provide complementary effects on risk reduction in cardiovascular adverse events and renal disease. Although the efficacy of CCBs as antihypertensive agents has been adequately demonstrated, there have been concerns regarding the use of short acting dihydropyridines after acute myocardial infarction. There have also been questions about the role of CCBs with regards to other antihypertensive agents in renal disease. For example, differential effects of dihydropyridine and non-dihydropyridine CCBs may affect progression of renal disease and risk for diabetes. Certain precautions involving drug interactions are needed because of the effects of CCBs on the CYP450 enzyme systems. PMID- 17109614 TI - Bullous pemphigoid treatment review. AB - Bullous pemphigoid is an autoimmune skin blistering disorder that can present with several different degrees of severity. The treatment modality employed by the treating physician varies from localised topical therapy and anti inflammatory treatments with minimal side effects to immunosuppressive agents associated with significant adverse reactions. Deciding which therapy to use with a particular patient can be a challenge, and the treating physician must take into account the severity of disease, the overall medical condition of the patient and potential drug interactions. This article provides a comprehensive review of current medical therapies, as well as an overall approach to the patient with bullous pemphigoid. PMID- 17109615 TI - Trilostane in advanced breast cancer. AB - Antiestrogens, principally tamoxifen, and aromatase inhibitors have been used as the first- and second-line therapy in patients with advanced postmenopausal breast cancer for many years. However, some patients acquire resistance to these treatments and, at present, further endocrine treatment is achieved by merely substituting the current medication with a different antiestrogen or aromatase inhibitor. Trilostane offers an alternative endocrine treatment due to its unique mode of action. It is an allosteric modulator of the estrogen receptor and targets both the estrogen- and growth factor-dependent pathways through which estradiol stimulates cell proliferation. In clinical trials, trilostane has been shown to be an effective treatment for breast cancer in patients who have relapsed after receiving treatment with one or more forms of endocrine therapy. Ongoing and future clinical trials are examining the potential for the use of trilostane in premenopausal breast cancer, as well as in other malignancies such as prostate cancer. PMID- 17109616 TI - Solifenacin provides effective antimuscarinic therapy for the complete management of overactive bladder. AB - Solifenacin is an antimuscarinic agent, administered once daily, which has been newly approved for the treatment of overactive bladder (OAB). Solifenacin administered at 5- and 10-mg once-daily doses shows efficacy for all the symptoms of OAB in both 'wet' and 'dry' patients, including improvements in patient quality of life and satisfaction. These improvements are observed as early as week 2 of treatment and are maintained over 12-week and 1-year time periods, without being compromised by the age or gender of the patient. Solifenacin demonstrates a favourable tolerability profile, with mild dry mouth as the most common adverse event associated with its use, both at the 5- and 10-mg doses; this allows for flexibility in the dosing regimen, in which physicians can administer solifenacin 5 mg, with the option to safely increase the dose to 10 mg if necessary based on the severity of patient's symptoms. The favourable efficacy and safety profile of solifenacin, coupled with its dose flexibility, are consistent with solifenacin being a convenient treatment option for patients with OAB. PMID- 17109617 TI - Sibutramine for obesity in adolescents. PMID- 17109618 TI - Formoterol as a rescue medication for asthma. PMID- 17109619 TI - Cellular balance, genes, and the Huang Ti Nei Ching Su Wen. PMID- 17109620 TI - NRF2-dependent glutamate-L-cysteine ligase catalytic subunit expression mediates insulin protection against hyperglycemia- induced brain endothelial cell apoptosis. AB - Increased oxidative stress and susceptibility of brain endothelium are contributing factors in the development of central nervous system complications in neuro-degenerative disorders in diabetes, Alzheimer's and Parkinson's disease. The molecular mechanisms underpinning the vulnerability of brain endothelial cells to chronic oxidative challenge have not been elucidated. Here, we investigated the oxidative susceptibility of human brain endothelial cells (IHEC) to chronic hyperglycemic stress and insulin signaling and cytoprotection. Chronic hyperglycemia exacerbated IHEC apoptosis in accordance with exaggerated cytosolic and mitochondrial glutathione and protein-thiol redox imbalance, and actin/Keap-1 S-glutathionylation. Insulin attenuated hyperglycemia-induced apoptosis via restored cytosolic and mitochondrial redox. Insulin stimulated glutamate-L cysteine ligase (GCL) activity by activation of phosphatidylinositol 3-kinase (PI3K)/Akt/mTOR signaling, increased serine phosphorylation and nuclear translocation of nuclear NF-E2-related factor 2 (Nrf2), and upregulation of Nrf2 dependent GCL-catalytic (GCLc) subunit expression. Expression of the GCL modulatory subunit (GCLm) was unchanged. Inhibitors of insulin receptor tyrosine kinase, PI3K, Akt and mTOR abrogated insulin-induced Nrf2-mediated GCLc expression, redox balance, and IHEC survival. Collectively, these results demonstrate that human brain endothelial cells exhibit vulnerability to hyperglycemic stress which is associated with marked cytosolic and mitochondrial redox shifts. Activation of insulin signaling through PI3K/Akt/mTOR/Nrf2/ GCLc pathway affords significant cell protection by maintaining cellular redox balance. PMID- 17109621 TI - Exercise preconditioning reduces brain damage and inhibits TNF-alpha receptor expression after hypoxia/reoxygenation: an in vivo and in vitro study. AB - Exercise reduces ischemia and reperfusion injury in rat stroke models. We investigated whether gradual increases in tumor necrosis factor-alpha (TNF-alpha) reported during exercise down-regulates expression of TNF-alpha receptors I and II (TNFRI and II) in stroke, leading to reduced brain damage. Adult male Sprague Dawley rats were subjected to 30 minutes of exercise on a treadmill each day for 3 weeks. Then, stroke was induced by a 2-hour middle cerebral artery (MCA) occlusion using an intra-luminal filament. Expressions of TNFRI and II mRNA in the brain were detected using a real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Protein expressions of TNFRI and II were determined by enzyme linked immunoabsorbant assay (ELISA) in serum and brain homogenates. Spatial distribution of TNF-alpha receptors in brain regions was determined with immunocytochemistry. In human umbilical vein endothelial cells (HUVEC), we addressed the causal effect of TNF-alpha pretreatment on TNF I and II expression using ELISA and real-time PCR. In exercised rats after stroke, brain infarct was significantly (p<0.01) reduced in the entire MCA supplied regions, associated with a mild expression of TNFRI and II mRNA and protein. The TNF-alpha receptors were restricted to the ischemic core. In contrast, a robust expression of TNFRI and II molecules was found in non-exercised rats subjected to similar ischemia/reperfusion insults. An in vitro study revealed a causal link between TNF-alpha pretreatment and reduced cellular expression of TNF-alpha receptors under hypoxic/reoxygenated conditions. Our results suggest that reduced-brain damage in ischemic rats after exercise preconditioning may be attributable to the reduced expression of TNF-alpha receptors. Chronically increased TNF-alpha expression was also found to reduce TNFI and II responding to acute ischemia/reperfusion insult. PMID- 17109622 TI - Dehydroepiandrosterone treatment alters lipid/phospholipid profiles of rat brain and liver mitochondria. AB - Stimulation of mitochondrial function following treatment with dehydroepiandrosterone (DHEA) has been demonstrated. Since the activity of several electron transport chain components is dependent on specific lipid/phospholipid components, we examined the effects of DHEA treatment (0.1-2.0 mg/kg body weight for 7 consecutive days) on lipid/phospholipids profiles of rat brain and liver mitochondria. In the brain mitochondria, contents of both total phospholipids (TPL) and cholesterol (CHL) increased. The major effect on phospholipids profile was increase in the contents of lysophospholipids (Lyso) and sphingomyelin (SPM) component followed by phosphatidylinositol (PI) and phosphatidylserine (PS). The contents of phosphatidylcholine (PC), phosphatidylethanolamine (PE) and diphosphatidylglycerol (DPG) were not affected. At the higher dose (2.0 mg) the observed effects declined. The TPL and CHL contents of liver mitochondria were generally unchanged by DHEA treatment. Under this condition the content of PI and PS increased. The contents of other phospholipid components were not changed. Our results suggest that the observed changes may complement the function of electron transport chain components. PMID- 17109623 TI - Derivation of Motor Neurons from three Clonal Human Embryonic Stem Cell Lines. AB - Human embryonic stem cells (hESC) demonstrate a remarkable proliferative and developmental potential and thus have huge therapeutic potential. To direct the differentiation of hESC to a specific lineage of high purity for cell transplantation is highly desirable. Here we describe a modified in vitro procedure to direct differentiation of three clonal hESC lines, hES 3.1, hES 3.2 and hES 3.3 efficiently to spinal motor neurons by using various differentiation factors namely retinoic acid (RA), sonic hedgehog (Shh), bone morphogenetic protein-2 (BMP-2) and Wnt3A. The highest number of motor neurons (58.0 +/- 7.6%) were obtained by an early treatment of embryoid bodies with a combination of RA + Shh from all the clonal hESC lines combined. The hES 3.1 line, however, produced relatively more motor neurons (69.5 +/- 11.8%) compared to other two hES clones, 3.2 (52.4 +/- 13.1%) and 3.3 (52.3 +/- 15.5%). Immunolocalisation studies revealed the expression of neuronal specific marker, beta omega-tubulin and motor neuron specific marker, HB9/HLXB9 in all the three hESC clones after 45 days of differentiation. The RT-PCR analyses showed the presence of the neuron-specific genes. This modified differentiation protocol provides a mean of obtaining an enriched population of motor neurons from hESC for possible use in studies of lineage development, drug discovery and also as a potential cell therapy for motor neuron disease. PMID- 17109624 TI - Classic beta-amyloid deposits cluster around large diameter blood vessels rather than capillaries in sporadic Alzheimer's disease. AB - Various hypotheses could explain the relationship between beta-amyloid (Abeta) deposition and the vasculature in Alzheimer's disease (AD). Amyloid deposition may reduce capillary density, affect endothelial cells of blood vessels, result in diffusion from blood vessels, or interfere with the perivascular clearance mechanism. Hence, the spatial pattern of the classic ('cored') type of Abeta deposit was studied in the upper laminae (I,II/III) of the superior frontal gyrus in nine cases of sporadic AD (SAD). Sections were immunostained with antibodies against Abeta and with collagen IV to study the relationships between the spatial distribution of the classic deposits and the blood vessel profiles. Both the classic deposits and blood vessel profiles were distributed in clusters. In all cases, there was a positive spatial correlation between the clusters of the classic deposits and the larger diameter (>10 microm) blood vessel profiles and especially the vertically penetrating arterioles. In only 1 case, was there a significant spatial correlation between the clusters of the classic deposits and the smaller diameter (<10 microm) capillaries. There were no negative correlations between the density of Abeta deposits and the smaller diameter capillaries. In 9/11 cases, the clusters of the classic deposits were significantly larger than those of the clusters of the larger blood vessel profiles. In addition, the density of the classic deposits declined as a negative exponential function with distance from a vertically penetrating arteriole. These results suggest that the classic Abeta deposits cluster around the larger blood vessels in the upper laminae of the frontal cortex. This aggregation could result from diffusion of proteins from blood vessels or from overloading the system of perivascular clearance from the brain. PMID- 17109625 TI - Neuronal semaphorins regulate a primary immune response. AB - Semaphorins are involved in a wide range of biological processes, including axon guidance, neuronal migration, angiogenesis, cardio- and osteo-genesis. Recently they have also been found to be important for immune response. Sema3A reduces the activation of T cells through its cell-surface receptors, including members of the neuropilin and plexin families. By contrast, Sema4D (CD100), which is expressed on the surface of T, B and dendritic cells, increases B cell and dendritic cell function using either plexin B1 or CD72 as receptors. The transmembrane protein Sema4A is involved in the activation of immune cells through interactions with Tim-2. Emerging evidence also indicates that additional semaphorins and related molecules seem to function in the reciprocal stimulation of T cells and antigen-presenting cells (APCs). This paper discusses the functions of these semaphorins in the immune system, focusing on their roles in T cell-APC interactions. PMID- 17109626 TI - Central nervous system circuitry and peripheral neural sympathetic activity responsible for essential hypertension. AB - Both clinical and experimental studies dealing with patients affected by idiopathic or essential hypertension (EH) are devoted to the great deal of physiological, pharmacological and pathological as well as therapeutical issues of EH. However, most articles devoted to EH do not refer to the central nervous system mechanisms underlying this disease and the channels which allow that these mechanisms are funneled to the peripheral autonomic nervous system and trigger this cardiovascular disorder. In the present review article we attempted to reach this target devoted to the central nervous system circuitry involved in the cardiovascular pathophysiology. We postulated that EH depends on the predominance of the binomial A5 noradrenergic (NA) nucleus + median raphe serotonergic (5-HT) nucleus over the (A6)-NA + dorsal raphe-5HT nuclei. This hypothesis receives additional support from our results obtained throughout the neuropharmacological therapy of this type of neurophysiological disorder. Our therapeutical strategy is addressed to enhance the activity of the (A6)-NA + dorsal raphe-5HT binomial circuitry. PMID- 17109627 TI - Hormonal regulation of the Menkes and Wilson copper-transporting ATPases in human placental Jeg-3 cells. AB - Copper deficiency during pregnancy results in early embryonic death and foetal structural abnormalities including skeletal, pulmonary and cardiovascular defects. During pregnancy, copper is transported from the maternal circulation to the foetus by mechanisms which have not been clearly elucidated. Two copper transporting ATPases, Menkes (ATP7A; MNK) and Wilson (ATP7B; WND), are expressed in the placenta and both are involved in placental copper transport, as copper accumulates in the placenta in both Menkes and Wilson disease. The regulatory mechanisms of MNK and WND and their exact role in the placenta are unknown. Using a differentiated polarized Jeg-3 cell culture model of placental trophoblasts, MNK and WND were shown to be expressed within these cells. Distinct roles for MNK and WND are suggested on the basis of their opposing responses to insulin. Insulin and oestrogen increased both MNK mRNA and protein levels, altered the localization of MNK towards the basolateral membrane in a copper-independent manner, and increased the transport of copper across this membrane. In contrast, levels of WND were decreased in response to insulin, and the protein was located in a tight perinuclear region, with a corresponding decrease in copper efflux across the apical membrane. These results are consistent with a model of copper transport in the placenta in which MNK delivers copper to the foetus and WND returns excess copper to the maternal circulation. Insulin and oestrogen stimulate copper transport to the foetus by increasing the expression of MNK and reducing the expression of WND. These data show for the first time that MNK and WND are differentially regulated by the hormones insulin and oestrogen in human placental cells. PMID- 17109628 TI - Importin alpha1 is involved in the nuclear localization of Zac1 and the induction of p21WAF1/CIP1 by Zac1. AB - Zac1, a novel seven-zinc-finger transcription factor, preferentially binds GC rich DNA elements and has intrinsic transactivation activity. To date, the NLS (nuclear localization signal) of Zac1 has not been empirically determined. We generated a series of EGFP (enhanced green fluorescence protein)-tagged deletion mutants of Zac1 and examined their subcellular localization, from which we defined two NLSs within the DNA-binding (or zinc-finger) domain. Fusion proteins consisting of the two EGFP-tagged zinc-finger clusters (zinc finger motifs 1-3 and 4-7) were located exclusively in the nucleus, demonstrating that each of the zinc-finger clusters is sufficient for nuclear localization. Physical interactions between these two zinc-finger clusters and importin alpha1 were demonstrated using an in vitro glutathione S-transferase pull-down assay. Finally, our results indicate that the association of Zac1 with importin alpha1 is also involved in regulating the transactivation activity of Zac1 on the p21WAF1/CIP1 gene and protein expression. PMID- 17109630 TI - Mathematical model for NF-kappaB-driven proliferation of adult neural stem cells. AB - Neural stem cells (NSCs) are early precursors of neuronal and glial cells. NSCs are capable of generating identical progeny through virtually unlimited numbers of cell divisions (cell proliferation), producing daughter cells committed to differentiation. Nuclear factor kappa B (NF-kappaB) is an inducible, ubiquitous transcription factor also expressed in neurones, glia and neural stem cells. Recently, several pieces of evidence have been provided for a central role of NF kappaB in NSC proliferation control. Here, we propose a novel mathematical model for NF-kappaB-driven proliferation of NSCs. We have been able to reconstruct the molecular pathway of activation and inactivation of NF-kappaB and its influence on cell proliferation by a system of nonlinear ordinary differential equations. Then we use a combination of analytical and numerical techniques to study the model dynamics. The results obtained are illustrated by computer simulations and are, in general, in accordance with biological findings reported by several independent laboratories. The model is able to both explain and predict experimental data. Understanding of proliferation mechanisms in NSCs may provide a novel outlook in both potential use in therapeutic approaches, and basic research as well. PMID- 17109629 TI - Functional properties of multiple isoforms of human divalent metal-ion transporter 1 (DMT1). AB - DMT1 (divalent metal-ion transporter 1) is a widely expressed metal-ion transporter that is vital for intestinal iron absorption and iron utilization by most cell types throughout the body, including erythroid precursors. Mutations in DMT1 cause severe microcytic anaemia in animal models. Four DMT1 isoforms that differ in their N- and C-termini arise from mRNA transcripts that vary both at their 5'-ends (starting in exon 1A or exon 1B) and at their 3'-ends giving rise to mRNAs containing (+) or lacking (-) the 3'-IRE (iron-responsive element) and resulting in altered C-terminal coding sequences. To determine whether these variations result in functional differences between isoforms, we explored the functional properties of each isoform using the voltage clamp and radiotracer assays in cRNA-injected Xenopus oocytes. 1A/IRE+-DMT1 mediated Fe2+-evoked currents that were saturable (K(0.5)(Fe) approximately 1-2 microM), temperature dependent (Q10 approximately 2), H+-dependent (K(0.5)(H) approximately 1 muM) and voltage-dependent. 1A/IRE+-DMT1 exhibited the provisional substrate profile (ranked on currents) Cd2+, Co2+, Fe2+, Mn2+>Ni2+, V3+>>Pb2+. Zn2+ also evoked large currents; however, the zinc-evoked current was accounted for by H+ and Cl- conductances and was not associated with significant Zn2+ transport. 1B/IRE+-DMT1 exhibited the same substrate profile, Fe2+ affinity and dependence on the H+ electrochemical gradient. Each isoform mediated 55Fe2+ uptake and Fe2+-evoked currents at low extracellular pH. Whereas iron transport activity varied markedly between the four isoforms, the activity for each correlated with the density of anti-DMT1 immunostaining in the plasma membrane, and the turnover rate of the Fe2+ transport cycle did not differ between isoforms. Therefore all four isoforms of human DMT1 function as metal-ion transporters of equivalent efficiency. Our results reveal that the N- and C-terminal sequence variations among the DMT1 isoforms do not alter DMT1 functional properties. We therefore propose that these variations serve as tissue-specific signals or cues to direct DMT1 to the appropriate subcellular compartments (e.g. in erythroid cells) or the plasma membrane (e.g. in intestine). PMID- 17109631 TI - High-potentiality preliminary selection criteria and transformation time dependent factors analysis for establishing Epstein-Barr virus transformed human lymphoblastoid cell lines. AB - Infection of freshly isolated and cryopreserved lymphocytes with Epstein-Barr virus (EBV) leads to the establishment of human B lymphoblastoid cell lines (LCLs). Techniques for optimal infection of the lymphocytes are vital for the establishment of a human biobank. The present study found that more than half (58 86%) of such established LCLs had transport times of less than 48 h, cell densities exceeding 10(6) cells/ml and cell viabilities greater than 90%. After EBV infection, 3306 freshly isolated lymphocytes required 30.0 +/- 0.1 days to become LCLs. Conversely, 1210 cryopreserved lymphocytes required 36.2 +/- 0.4 days. Cell density and viability of the culture affected transformation time in freshly isolated lymphocytes. On the other hand, blood transport time, cryopreservation time and initial cell viability were major factors in cryopreserved specimens. These results contribute to general information concerning the establishment of a human biobank for EBV infected cells. PMID- 17109632 TI - Cell transplantation with a catheter-based approach: an efficient method for the treatment of heart failure with multiple lesions. AB - Cell transplantation is emerging as a potential therapeutic approach for the treatment of heart failure. At present, popular methods of cell delivery may not be efficient in perfusing cells through the whole myocardium. We have developed a novel catheter-based method for global transplantation of cells. Heart failure was induced in rabbit by intravenous administration of doxorubicin. Autologous bone marrow mononuclear cells were transplanted into failing hearts via the root of the aorta. Bilateral sinus aortae and coronary arteries were visualized by angiography during the cell transplantation procedure; there was no intraprocedural death. Four weeks after cell transplantation, there was an improvement in the mean left ventricular ejection fraction from baseline 72.13% to 81.54% (P = 0.034). Transplanted cells were observed throughout the cardiac layers of left and right ventricles. In conclusion, cell transplantation through the root of the aorta is a useful approach to globally supply cells into the heart. PMID- 17109633 TI - Time- and concentration-dependent effects of resveratrol in HL-60 and HepG2 cells. AB - Resveratrol, a phytochemical present in grapes, has been demonstrated to inhibit tumourigenesis in animal models. However, the specific mechanism by which resveratrol exerts its anticarcinogenic effect has yet to be elucidated. In the present study, the inhibitory effects of resveratrol on cell proliferation and apoptosis were evaluated in the human leukaemia cell line HL-60 and the human hepatoma derived cell line HepG2. We found that after a 2 h incubation period, resveratrol inhibited DNA synthesis in a concentration-dependent manner. The IC50 value was 15 microm in both HL-60 and HepG2 cells. When the time of treatment was extended, an increase in IC50 value was observed; for example, at 24 h the IC50 value was 30 microm for HL-60 cells and 60 microm for HepG2 cells. Flow cytometry revealed that cells accumulated in different phases of the cell cycle depending on the resveratrol concentration. Furthermore, an increase in nuclear size and granularity was observed in the G1 and S phases of HL-60 treated and HepG2 treated cells. Apoptosis was also stimulated by resveratrol in a concentration dependent manner in HL-60 and HepG2 cells. In conclusion, resveratrol inhibits cell proliferation in a concentration- and time-dependent manner by interfering with different stages of the cell cycle. Furthermore, resveratrol treatment causes stimulation of apoptosis as well as an increase in nuclear size and granularity. PMID- 17109634 TI - Cytomics - importance of multimodal analysis of cell function and proliferation in oncology. AB - Cancer is a highly complex and heterogeneous disease involving a succession of genetic changes (frequently caused or accompanied by exogenous trauma), and resulting in a molecular phenotype that in turn results in a malignant specification. The development of malignancy has been described as a multistep process involving self-sufficiency in growth signals, insensitivity to antigrowth signals, evasion of apoptosis, limitless replicative potential, sustained angiogenesis, and finally tissue invasion and metastasis. The quantitative analysis of networking molecules within the cells might be applied to understand native-state tissue signalling biology, complex drug actions and dysfunctional signalling in transformed cells, that is, in cancer cells. High-content and high throughput single-cell analysis can lead to systems biology and cytomics. The application of cytomics in cancer research and diagnostics is very broad, ranging from the better understanding of the tumour cell biology to the identification of residual tumour cells after treatment, to drug discovery. The ultimate goal is to pinpoint in detail these processes on the molecular, cellular and tissue level. A comprehensive knowledge of these will require tissue analysis, which is multiplex and functional; thus, vast amounts of data are being collected from current genomic and proteomic platforms for integration and interpretation as well as for new varieties of updated cytomics technology. This overview will briefly highlight the most important aspects of this continuously developing field. PMID- 17109635 TI - Protein encoded by HSV-1 stimulation-related gene 1 (HSRG1) interacts with and inhibits SV40 large T antigen. AB - Herpes simplex virus (HSV)-1 stimulation-related gene 1 (HSRG1) protein expression is induced in HSV-1 infected cells. We found that HSRG1 interacts with SV40 large T antigen (LT) in yeast two-hybrid assay and bimolecular fluorescence complementation (BiFC) assay. This interaction alters LT's regulation of the SV40 promoter and its ability to influence the cell cycle. Choramphenicol acetyl transferase (CAT) assays revealed that initiation of gene transcription by LT is changed by HSRG1 expression. HSRG1 inhibits the ability of LT to activate SV40 late gene transcription. Further data indicate that the ability of LT protein to stimulate S-phase entry is also inhibited by the expression of HSRG1. The results of a colony-forming assay suggested that expression of HSRG1 in cells transfected by LT gene decreased the rate of colony formation. Yeast two-hybrid beta galactosidase assay revealed that amino acid residues 132-450 in LT bind HSRG1. PMID- 17109636 TI - Different isoforms of the Wilms' tumour protein WT1 have distinct patterns of distribution and trafficking within the nucleus. AB - The Wilms' tumour suppressor gene WT1 encodes multiple isoforms of a transcription factor essential for correct mammalian urogenital development. Maintenance of the correct isoform ratio is critical. In humans, perturbation of this ratio causes Frasier syndrome, which is characterized by developmental defects of the kidney and urogenital tract. Different WT1 isoforms are thought to regulate transcription and participate in mRNA processing, functions reflected by a complex sub-nuclear distribution. However, the role of individual WT1 isoforms remains unclear and pathways leading to WT1 sub-nuclear localization are completely unknown. Here we use cells expressing green fluorescent protein-tagged WT1 to demonstrate that the two major WT1 isoforms occupy separate and dynamic intranuclear locations in which one isoform, WT1+KTS, preferentially associates with the nucleolus. The alternatively spliced zinc finger region is found to be critical for the initial sub-nuclear separation of WT1 isoforms, but interactions between different isoforms influence the sub-nuclear distribution of WT1. We illustrate how disruption of WT1 nuclear distribution might result in disease. This study contributes to the emerging picture of intranuclear protein trafficking. PMID- 17109637 TI - Role of FGF1, FGF2 and FGF7 in the development of the pancreas from control and streptozotocin-treated hamsters. AB - Although progress has been made with respect to the growth and transcription factors implicated in pancreatic development, many questions remain unsolved. It has been established that during embryonic life, both endocrine and acinar cells are derived from pancreatic epithelial precursor cells. Growth factors control the proliferation of precursor cells and their ability to differentiate into mature cells, both in pre-natal and in early post-natal life. Pancreatic development during the early post-natal period is an area of great interest for many scientists. In this study we have examined the structure characteristics, functional and proliferative activity of control and diabetic hamster pancreatic ductal, exocrine and beta cells, following treatment with FGFs 1, 2 and 7 in vitro. Light and electron microscopic studies indicated active synthetic processes in these cells under the influence of the investigated FGFs. In our experimental model of diabetes, the labelling index of the cells was significantly higher than in corresponding control groups of hamsters. We established that FGF2 at a concentration of 10 ng/l was responsible for the most prominent effect on ductal cells and beta cells in the diabetic groups. FGF1 at a concentration of 10 ng/l displayed the highest stimulatory effect on exocrine cells in the diabetic groups at post-natal day 10. Taken together these data strongly suggest that FGF1 and FGF2 induce proliferation of pancreatic epithelial cells during the early post-natal period whereas FGF7 is not strictly specific for pancreatic cell proliferation. PMID- 17109638 TI - Sensitivity of cells to apoptosis induced by iron deprivation can be reversibly changed by iron availability. AB - We tested the effect of iron deprivation on cell death induction in human Raji cells pre-adapted to differing availability of extracellular iron. Iron deprivation was achieved by incubation in a defined iron-free medium. Original Raji cells have previously been adapted to long-term culture in a defined medium with 5 microg/ml of iron-saturated human transferrin as a source of iron. Raji/lowFe cells were derived from original Raji cells by subsequent adaptation to culture in the medium with 50 microm ferric citrate as a source of iron. Raji/lowFe-re cells were derived from Raji/lowFe cells by re-adaptation to the transferrin-containing (5 microg/ml) medium. Iron deprivation induced cell death in both Raji cells and Raji/lowFe-re cells; that is, cells pre-adapted to a near optimum source of extracellular iron (5 microg/ml of transferrin). However, Raji/lowFe cells preadapted to a limited source of extracellular iron (50 microm ferric citrate) became resistant to the induction of cell death by iron deprivation. We demonstrated that cell death induction by iron deprivation in Raji cells correlates with the activation of executioner caspase-3 and the cleavage of caspase-3 substrate, poly-ADP ribose polymerase. Two other executioner caspases, caspase-7 and caspase-6, were not activated. Taken together, we suggest that in human Raji cells, iron deprivation induces apoptotic cell death related to caspase-3 activation. However, the sensitivity of the cells to death induction by iron deprivation can be reversibly changed by extracellular iron availability. The cells pre-adapted to a limited source of extracellular iron became resistant. PMID- 17109639 TI - gamma-Tocotrienol inhibits ErbB3-dependent PI3K/Akt mitogenic signalling in neoplastic mammary epithelial cells. AB - The antiproliferative effects of gamma-tocotrienol are associated with suppression in epidermal growth factor (EGF)-dependent phosphatidylinositol-3 kinase (PI3K)/PI3K-dependent kinase-1 (PDK-1)/Akt mitogenic signalling in neoplastic mammary epithelial cells. Studies were conducted to investigate the direct effects of gamma-tocotrienol treatment on specific components within the PI3K/PDK-1/Akt mitogenic pathway. +SA cells were grown in culture and maintained in serum-free media containing 10 ng/ml EGF as a mitogen. Treatment with 0-8 microm gamma-tocotrienol resulted in a dose-responsive decrease in the +SA cell growth and a corresponding decrease in phospho-Akt (active) levels. However, gamma-tocotrienol treatment had no direct inhibitory effect on Akt or PI3K enzymatic activity, suggesting that the inhibitory effects of gamma-tocotrienol occur upstream of PI3K, possibly at the level of the EGF-receptor (ErbB1). Additional studies were conducted to determine the effects of gamma-tocotrienol on ErbB receptor activation. Results showed that gamma-tocotrienol treatment had little or no effect on ErbB1 or ErbB2 receptor tyrosine phosphorylation, a prerequisite for substrate interaction and signal transduction, but did cause a significant and progressive decrease in the ErbB3 tyrosine phosphorylation. Because ErbB1 or ErbB2 receptors form heterodimers with the ErbB3 receptor, and ErbB3 heterodimers have been shown to be the most potent activators of PI3K, these findings strongly suggest that the antiproliferative effects of gamma tocotrienol in neoplastic +SA mouse mammary epithelial cells are mediated by a suppression in ErbB3-receptor tyrosine phosphorylation and subsequent reduction in PI3K/PDK-1/Akt mitogenic signalling. PMID- 17109640 TI - Possibility of selection of chondrogenic progenitor cells by telomere length in FGF-2-expanded mesenchymal stromal cells. AB - Telomere length plays an important role in regulating the proliferative capacity of cells, and serves as a marker for cell cycle history and also for their remaining replicative potential. Mesenchymal stromal cells (MSC) are known to be a significant cell source for therapeutic intervention and tissue engineering. To investigate any possible limitations in the replicative potential and chondrogenic differentiation potential of fibroblast growth factor-2-expanded MSCs (FGF(+)MSC), these cells were differentiated at various population doublings (PDs), and telomere length and telomerase activity were measured before and after differentiation. FGF(+)MSC cultured at a relatively low density maintained proliferation capability past more than 80 PD and maintained chondrogenic differentiation potential up to at least 46 PD and long telomeres up to 105 PD, despite expressing low levels of telomerase activity. Interestingly, upon chondrogenic differentiation of these cells, telomeres showed a remarkable reduction in length. This shortening was more extensive when FGF(+)MSC of higher PD levels were differentiated. These findings suggest that telomere length may be a useful genetic marker for chondrogenic progenitor cells. PMID- 17109641 TI - Assessment of pluripotency and multilineage differentiation potential of NTERA-2 cells as a model for studying human embryonic stem cells. AB - Embryonal carcinoma cells are pluripotent stem cells derived from teratocarcinomas and are considered to be the malignant counterparts of human embryonic stem cells. As there are few reliable experimental systems available to study the molecular mechanisms governing normal embryogenesis, well-characterized human embryonal carcinoma stem cell lines may provide a robust and simple model to study certain aspects of pluripotency and cellular differentiation. Here, we have analysed NTERA-2 cL.D1 cells at molecular and cellular levels during expansion and differentiation, via formation of cell aggregates similar to embryoid bodies in embryonic stem cells. Thus, human embryonal carcinoma cells may provide a valuable insight into cell fate determination, into the embryonic ectoderm, mesoderm and endoderm and their downstream derivatives. PMID- 17109642 TI - Syringolin A, a new plant elicitor from the phytopathogenic bacterium Pseudomonas syringae pv. syringae, inhibits the proliferation of neuroblastoma and ovarian cancer cells and induces apoptosis. AB - Syringolin A is a new plant elicitor produced by the plant pathogen Pseudomonas syringae pv. syringae. The goal of this study was to investigate whether syringolin A exhibits anti-proliferative properties in cancer cells. The treatment of human neuroblastoma (NB) cells (SK-N-SH and LAN-1) and human ovarian cancer cells (SKOV3) with syringolin A (0-100 microm) inhibited cell proliferation in a dose-dependent manner. The IC(50) (50% inhibition) for each cell line ranged between 20 microm and 25 microm. In SK-N-SH cells, the treatment with 20 microm syringolin A led to a rapid (24 h) increase of the apoptosis associated tumour suppressor protein p53. In addition, we found that the treatment of SK-N-SH cells caused severe morphological changes after 48 h such as rounding of cells and loss of adherence, both conditions observed during apoptosis. The induction of apoptosis by syringolin A was confirmed by both poly (ADP-ribose) polymerase (PARP) cleavage and annexin V assay. Taken together, we show for the first time that the natural product syringolin A exhibits anti proliferative activity and induces apoptosis. Syringolin A and structurally modified syringolin A derivatives may serve as new lead compounds for the development of novel anticancer drugs. PMID- 17109643 TI - Establishment and characterization of new mammary adenocarcinoma cell lines derived from double transgenic mice expressing GFP and neu oncogene. AB - A new murine cell line, named GFPneu, was established from a mammary adenocarcinoma arising in double transgenic MMTVneu x CMV-GFP mice. Breast tumours develop in 100% of females after 2 months latency, as a result of the over-expression of the activated rat neu oncogene in the mammary glands. All tissues, and in particular the breast tumours, express the GFP protein. This cell line was tumorigenic when inoculated into nude mice and the derived tumours showed the same histological features as the primaries from which they were isolated. Their histopathology reproduces many characteristics of human breast adenocarcinomas, in particular their ability to metastasize. The GFP marker allows us to visualize the presence of lung metastases in fresh tissues immediately, to confirm the histopathology. From a lung metastatic fluorescent nodule, we derived a further cell line, named MTP-GFP, which we also characterized. These two cell lines could be useful to study the role played by the neu oncogene in the maintenance of the transformed phenotype, in the metastatic process, to test novel therapeutic strategies to inhibit primary tumour growth and to observe the generation of distant metastases. PMID- 17109644 TI - Hypericum caprifoliatum (Guttiferae) Cham. & Schltdl.: a species native to South Brazil with antidepressant-like activity. AB - In this work, previously published and unpublished results on biological activity of Hypericum caprifoliatum, a native species to South Brazil, are presented. Lipophilic extracts obtained from this species showed an antidepressant-like activity in mice and rat forced swimming test. Results from in vivo experiments suggest an effect on the dopaminergic transmission. Besides that, in vitro experiments demonstrated that the extract and its main component (a phloroglucinol derivative) inhibit monoamine uptake in a concentration-dependent manner, more potently to dopamine, but this effect is not related to direct binding at the uptake sites. It was also observed that a 3-day treatment with lipophilic extract prevents stress-induced corticosterone rise in mice frontal cortex but not in plasma. The lipophilic and methanolic H. caprifoliatum extracts also demonstrated antinociceptive effect, which seems to be indirectly mediated by the opioid system. These results indicate that H. caprifoliatum presents a promising antidepressant-like effect in rodents which seems to be related to a mechanism different from that of other classes of antidepressants. PMID- 17109645 TI - How to deal with multiple endpoints in clinical trials. AB - Multiple endpoints are common in clinical trials. This article discusses statistical methods that can be applied to control the rate of false positive conclusions at an acceptable level. The considered methods include the Bonferroni adjustment and related methods, the intersection-union test, ordered hypotheses and gatekeeper procedures, composite endpoints and global assessment measures, closed testing procedures, and combinations of different approaches. PMID- 17109646 TI - Prevention of sudden cardiac death by n-3 polyunsaturated fatty acids. AB - There were already several epidemiologic studies that showed eating fish frequently seemed to reduce deaths from coronary heart disease. There were also observational and clinical trials, which more specifically showed that the reduction in cardiovascular deaths from eating fish were largely the result of n 3 polyunsaturated fatty acids in fish oil for the prevention of sudden cardiac death (SCD). This led me to perform a clinical trial in which all subjects had an implanted cardioverter-defibrillator and were at very high risk of SCD. The results of this study and the mechanisms by which the n-3 fish oil fatty acids prevent fatal cardiac arrhythmias will be the subject of this review. PMID- 17109647 TI - Understanding the oestrogen action in experimental and clinical atherosclerosis. AB - Whereas hormone replacement/menopause therapy (HRT) in postmenopausal women increases the coronary artery risk, epidemiological studies (protection in premenopaused women) suggest and experimental studies (prevention of the development of fatty streaks in animals) demonstrate a major atheroprotective action of oestradiol (E2). The understanding of the deleterious and beneficial effects of oestrogens is thus required. The immuno-inflammatory system plays a key role in the development of fatty streak deposit as well as in the rupture of the atherosclerotic plaque. Whereas E2 favours an anti-inflammatory effect in vitro (cultured cells), it rather elicits in vivo a proinflammation at the level of several subpopulations of the immuno-inflammatory system, which could contribute to plaque destabilization. Endothelium is another important target for E2, as it potentiates endothelial NO and prostacyclin production, thus promoting the beneficial effects as vasorelaxation and inhibition of platelet aggregation. Prostacyclin, but not NO, appears to be involved in the atheroprotective effect of E2. E2 also accelerates endothelial regrowth, thus favouring vascular healing. Finally, most of these effects of E2 are mediated by oestrogen receptor alpha, and are independent of oestrogen receptor beta. In summary, a better understanding of the mechanisms of oestrogen action not only on the normal and atheromatous arteries, but also on innate and adaptive immune responses is required and should help to optimize the prevention of cardiovascular disease after menopause. These mouse models should help to screen existing and future selective oestrogen receptor modulators. PMID- 17109648 TI - The cannabinoid system and its pharmacological manipulation--a review, with emphasis upon the uptake and hydrolysis of anandamide. AB - Although cannabis has been used both recreationally and for medicinal purposes since ancient times, it was not until the 1990s that the receptors responsible for many of the actions of Delta(9)-tetrahydrocannabinol, the main psychoactive ingredient of cannabis, were cloned. Since then, our knowledge of the endogenous cannabinoid system, its physiology, pharmacology and therapeutic potential have expanded enormously. In the present review, the cannabinoid system is described, with particular emphasis on the mechanisms of removal and metabolism of the endocannabinoid signalling molecule anandamide. The current literature shows that cells can accumulate anandamide, and that this process can be disrupted pharmacologically, but that the nature of the mechanism(s) involved remains a matter of some debate. The main enzyme for the hydrolysis of anandamide, fatty acid amide hydrolase, is well characterized, and molecules selectively inhibiting this enzyme have potential therapeutic utility in a number of areas, in particular for the treatment of pain conditions. PMID- 17109649 TI - Clinical and biological consequences of transmetallation induced by contrast agents for magnetic resonance imaging: a review. AB - Gadolinium-based contrast agents (CAs) are widely used to enhance the contrast of images in magnetic resonance imaging procedures. Two categories of gadolinium chelates exist: the macrocyclic molecules where Gd3+ is caged in the pre organized cavity of the ligand and the linear molecules. Gadolinium chelates differ in their thermodynamic stability constants and in their kinetic stability. In general, macrocyclic chelates such as Gd-DOTA or Gd-HP-DO3A are more stable than linear molecules. Even among linear agents, differences can be found. There is increasing evidence that transmetallation can be found in vivo, in the case of certain CAs (especially linear chelates), with body cations such as zinc, calcium or iron. Furthermore, analytical interference with colorimetric determination of calcium has been clinically evidenced with two linear chelates, Gd-DTPA-BMA and Gd-DTPA-BMEA. Clinical cases of spurious hypocalcaemia have been reported with these molecules. Such interference with some colorimetric assays for calcium is clinically relevant in that it can lead to unnecessary and potentially harmful treatment for hypocalcaemia. PMID- 17109650 TI - Valvular heart disease in a patient taking benfluorex. AB - The authors describe a case of valvular heart disease in a 48-year-old woman receiving benfluorex (150 mg t.i.d. for 8 years) and leading to surgical mitral valve replacement. Examination showed severe dyspnea with severe mitral regurgitation associated with tricuspid regurgitation. No other common disease known to affect mitral valves nor intake of other drugs (ergot derivatives or appetite suppressant drugs) was found. The paper discusses the imputability of benfluorex, a drug used as an adjuvant in hypercholesterolemia, structurally related to amphetamines. PMID- 17109651 TI - Cassava-enriched diet is not diabetogenic rather it aggravates diabetes in rats. AB - Chronic intake of cassava has been thought to play a role in the pathogenesis of diabetes. We investigated the effects of dietary cassava (Manihot esculenta), which naturally contains cyanogenic glycosides, in the progression of diabetes mellitus in rats. Diabetes was induced by five mild doses of streptozotocin, in male Wistar rats which were fed a standard or cyanide-free cassava (CFC) diet containing or not containing exogenous cyanide with or without methionine. Methionine was employed to counterbalance the toxic effects of cyanide. During diabetes progression, we determined glycaemia and antioxidant status, by measuring vitamin C levels and activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and glutathione reductase (GSSG-Red). Feeding CFC diet did not induce diabetes in control rats; rather this diet, in diabetic animals, aggravated hyperglycaemia the severity of which was increased in these animals fed CFC diet, supplemented with cyanide. Addition of methionine curtailed the toxic effects of cyanide supplementation in CFC diet-fed diabetic animals. In standard diet-fed animals, the activities of SOD, GSH-Px and GSSG-Red were lower in diabetic rats than control rats. Interestingly, all of the CFC diets with or without cyanide or methionine, increased vitamin C levels and antioxidant enzyme activities in both control and diabetic animals. However, supplementing cyanide to CFC diet (without methionine) curtailed SOD and GSH-Px activities in diabetic rats. Our study shows that cassava diet containing cyanide is 'diabetes aggravating'. PMID- 17109652 TI - Evaluation of a surgical procedure to measure drug biliary excretion of rats in regulatory safety studies. AB - A surgical procedure was evaluated to allow bile collection from the freely moving male Sprague-Dawley rats for the assessment of drug biliary excretion during regulatory safety studies. A catheter was implanted into the bile duct to divert the bile flow via an exteriorized loop. Following recovery from the surgery and verification of normal hepatic function, the exteriorized catheter was sectioned to allow collection of the bile and replacement with a commercial bile salt solution. Approximately 80% of the catheterized animals (10 females and 10 males) had normal serum liver enzyme levels 2 days after surgery. Then, the effect of acute or repeated administrations of the immunosuppressant tacrolimus on the biliary excretion of 14C diazepam was studied to validate the technique. A first group of 12 rats received an intravenous injection of 10 mg/kg 14C-diazepam and the total and sequential amounts of diazepam excreted in the bile were measured over 72 h. Biliary excretion accounted for 80% of diazepam elimination. These rats were then given an oral administration of 3 mg/kg tacrolimus on days 7 and 8 followed by the same intravenous dose of 14C-diazepam. Another group of 10 catheterized rats was given 21 daily oral doses of 3 mg/kg tacrolimus followed by a single intravenous administration of 14C-diazepam. No significant changes in diazepam biliary excretion were observed following either acute or repeated administration of tacrolimus. This study demonstrates the feasibility of drug biliary excretion investigations under Good Laboratory Practices conditions as a complement to regulatory acute or repeated dose safety studies. PMID- 17109653 TI - Possible role of exogenous cAMP to improve vascular endothelial dysfunction in hypertensive rats. AB - The study has been designed to investigate the effect of 8-Br-cAMP, an activator of protein kinase A, in hypertension-induced vascular endothelial dysfunction. Rats were uninephroctomized and desoxycortisone acetate (DOCA) (40 mg/kg, s.c.) was administered to rats to produce hypertension (mean arterial blood pressure > 140 mmHg). Vascular endothelial dysfunction was assessed using isolated aortic ring preparation, electron microscopy of thoracic aorta and serum concentration of nitrite/nitrate. The expression of mRNA for p22phox and eNOS was assessed by using reverse transcriptase-polymerase chain reaction. Serum thiobarbituric acid reactive substances concentration and aortic superoxide anion concentration were estimated to assess oxidative stress. 8-Br-cAMP (5 mg/kg, i.p.) or atorvastatin (30 mg/kg, p.o.) prevented hypertension-induced attenuation of acetylcholine induced endothelium-dependent relaxation, impairment of vascular endothelial lining, decrease in expression of mRNA for endothelial nitric oxide synthase (eNOS), serum nitrite/nitrate concentration and increase in expression of mRNA for p22phox, superoxide anion and serum TBARS. The ameliorative effect of 8-Br cAMP was prevented by N-nitro-L-arginine methyl ester (25 mg/kg, i.p.) and glibenclamide (30 mg/kg, i.p.). It may be concluded that 8-Br-cAMP may stimulate expression and activity of eNOS and suppress expression of p22phox subunit of NADPH oxidase to reduce oxidative stress and subsequently improve vascular endothelial dysfunction. PMID- 17109654 TI - Ultrastructural clues for the potent therapeutic effect of melatonin on aging skin in pinealectomized rats. AB - Recently we have reported a significant reduction in the thickness of epidermis and epidermis + dermis in the back, abdominal and thoracic skin of the long-term pinealectomized rats and the potent therapeutic effect of melatonin on the pinealectomy-induced morphometric changes. The present study was aimed to determine the fine structure of the abdominal and thoracic skin in pinealectomized rats and the effect of melatonin on skin ultrastructure. Rats were pinealectomized or sham operated (control) for 6 months. Half of the pinealectomized rats were treated with 4 mg/kg melatonin during the last month of the experiment. Pinealectomy resulted in prominent ultrastructural changes in the skin. Epidermal atrophy, disorganization and cytological atypia were obvious. Tonofilament distribution was not uniform, and intercellular space was narrow. Nuclear irregularity and heterochromatin condensation were detected. Many mitochondria were irregular and edematous with increased translucence of the matrix, either partial or total destruction of crests and frequently the presence of vacuoles, myelin figures and dense bodies. Microprojections of basal cells into the dermis were observed. The dermis was thin, and collagenous fibers were loosely arranged. The epidermis in melatonin administered pinealectomized rats was obviously thicker than that of pinealectomized rats. The cells of each layers had characteristic morphological and ultrastructural features. Nuclear irregularity and heterochromatin condensation were not seen. Mitochondria were generally normal in ultrastructural appearance but rarely vacuoles and myelin figures were observed. The dermis was thick, and collagenous fibers were closely packaged. This paper provides an additional ultrastructural evidence that the damage to mitochondria is the major contributory factor to skin aging and that melatonin has potent therapeutic effects in reducing age-related changes via protecting fine structure of the skin. PMID- 17109655 TI - Synergistic inhibition of the enzymatic activity of aminopeptidase N by divalent metal ion chelators. AB - Membranes of HEK293 cells that were transfected with human aminopeptidase N (AP N, CD13, EC 3.4.11.2) and purified soluble porcine kidney AP-N were used to study inhibition of its enzyme activity by divalent cation chelators. Whereas pre incubation for 10 min with ethylenediaminetetraacetic acid (EDTA), did not or only weakly affected the enzyme activity, the bidentate chelator 1,10 phenanthroline produced a complete and concentration-dependent inhibition of AP N. The corresponding curves had Hill slopes of 2.50 +/- 0.23 and 2.73 +/- 0.01 for soluble and recombinant AP-N respectively. EDTA increased the potency of 1,10 phenanthroline till a limit, at which Hill slopes became close to unity. In the absence of EDTA, the inhibition by 1,10-phenanthroline was only weakly affected by the substrate concentration. On the other hand, competition between 1,10 phenanthroline and the substrate took place in the presence of EDTA. Similar findings were reported for the related metallopeptidase cystinyl aminopeptidase and point towards a model in which 1,10-phenanthroline inhibit enzyme activity by decreasing the free Zn2+ concentration. Moreover, EDTA is capable of removing a modulatory ion from an allosteric site at the enzyme, facilitating the direct interaction between 1,10-phenanthroline and the catalytic Zn2+. Compatible with this model, Ca2+ may bind to this allosteric site resulting in the potentiation of Zn2+-mediated re-activation of the enzyme activity in the presence of EDTA and 1,10-phenanthroline. PMID- 17109658 TI - The importance of getting treatment for headache right. PMID- 17109659 TI - Exenatide and pramlintide: new therapies for diabetes. PMID- 17109660 TI - Carbon monoxide poisoning. PMID- 17109661 TI - Oxidative stress and hypertension. PMID- 17109662 TI - Munchausen syndrome by proxy in pregnancy. PMID- 17109663 TI - Improved migraine management in primary care: results of a patient treatment experience study using zolmitriptan orally disintegrating tablet. AB - The 'Zomig Appropriate for Primary care' programme was developed to address the needs of primary care physicians (PCPs) to improve migraine management. As part of the programme, an international, open-label, 6-month clinical study was performed. The study included new and tangible outcome variables relevant to PCPs and recruited patients presenting in primary care with an established migraine diagnosis. Patients treated up to three migraine attacks per month with zolmitriptan orally disintegrating tablet (ODT) 2.5 mg. All other migraine attacks occurring during the study period were treated with the patient's usual migraine medication (including other triptans). Questionnaires were used to record patient treatment experiences at the study end. The primary end-point was the proportion of patients wanting to continue using zolmitriptan ODT. Some 595 patients treated 7171 migraine attacks with zolmitriptan ODT. Of the 504 patients who completed the 6-month questionnaire, 380 (75.4%) wished to continue using zolmitriptan ODT. The results of the study indicate that patient-orientated end points are more motivational and meaningful to physicians than traditional end points used in controlled clinical trials, allowing them to make informed decisions regarding migraine management. PMID- 17109664 TI - Antibody response after varicella vaccination in children treated with budesonide inhalation suspension or non-steroidal conventional asthma therapy. AB - We evaluated if budesonide inhalation suspension (BIS) reduces the immunogenicity of the varicella vaccine in paediatric patients with asthma. This open-label, parallel-group, cohort study included varicella-naive (disease and vaccine) children aged 12 months to 8 years with asthma requiring therapy. Patients who received > or = 4 weeks of asthma treatment with BIS 0.25-1 mg daily or non steroidal conventional asthma therapy (NSCAT) daily or as needed and met eligibility requirements received the varicella vaccine (Varivax) and continued the same asthma treatment for > or = 8 weeks postvaccination. Varicella-zoster virus (VZV) antibody levels were assessed before and 6 weeks after vaccination using a glycoprotein enzyme-linked immunosorbent assay (gpELISA). Adverse events (AEs) were assessed throughout the study. Antibody levels were analysed in 243 of 274 patients who were vaccinated and received treatment. After immunisation, the percentage of patients in each group achieving a 'protective' level of VZV antibody (> or = 5 gpELISA units/ml) was similar: 85% (129/151) in the BIS group and 90% (83/92) in the NSCAT group (relative risk = 0.95; 95% confidence interval 0.86-1.04). Eight patients in each group reported AEs related to varicella vaccination (primarily pyrexia, agitation and injection-site reactions). There were no cases of severe varicella in either group; one case of mild varicella like rash was reported in a 12-month-old child in the NSCAT group 11 days after vaccination. VZV antibody responses and tolerability to the live varicella vaccine in paediatric asthma patients treated with BIS vs. NSCAT were comparable, demonstrating that young children with asthma receiving nebulised BIS can be immunised effectively with Varivax. PMID- 17109665 TI - Oxidative stress in human in sustained and white coat hypertension. AB - Oxidative stress is thought to play a critical role in the pathogenesis of hypertension. Protein oxidation is defined here as the covalent modification of a protein induced either directly by reactive oxygen species or indirectly by reaction with secondary by-products of oxidative stress. The aim of our study was to evaluate the protein oxidation and to examine the function of the antioxidative system in sustained and white coat hypertensives (WCH) and compare with normotensives. This study was designed to investigate the protein oxidation parameters [protein carbonyls (PCOs)] in sustained hypertensives (17 males and 20 females) and WCH (18 males and 19 females). PCO and the endogenous antioxidant components protein thiol (P-SH), CuZn-superoxide dismutase (CuZn-SOD) and glutathione (GSH) were analysed using spectrophotometric and kinetic methods. Sustained hypertensive and WCH groups exhibited higher protein oxidation and lower P-SH, CuZn-SOD and GSH activities than normotensives. With regard to these parameters, there was no significant difference between sustained hypertensive and WCH groups. Blood pressure correlates positively with PCO groups and negatively with others. There exists an imbalance between oxidants and antioxidants in WCH because of the increase of oxidants associated with the decrease of antioxidant capacity. This may cause endothelial dysfunction just like in sustained hypertension. It may be necessary to add antioxidants to conventional antihypertensive therapy to balance the oxidative status in WCH. PMID- 17109666 TI - Correlation of plasma leptin and adiponectin with insulin sensitivity and beta cell function in children - the Taipei Children Heart Study. AB - To investigate the association between plasma leptin and adiponectin and insulin sensitivity in children, 580 school children (294 boys and 286 girls) with mean age of 13.3 years (12-16 years) were randomly selected from the Taipei Children Heart Study. Baseline measurements included body weight, body mass index (BMI), plasma glucose, insulin, proinsulin, leptin and adiponectin levels. Insulin resistance and beta-cell function were assessed using the method of homeostatic model, HOMA-IR and HOMA-beta, respectively. We found that girls had higher levels of plasma leptin, adiponectin and HOMA-beta than boys. There was no significant difference in HOMA-IR between boys and girls. Plasma leptin concentrations were positively correlated with body weight, BMI, insulin and proinsulin concentrations, HOMA-IR and HOMA-beta, whereas plasma adiponectin levels were inversely associated with body weight, BMI and proinsulin levels in both sexes. In girls, adiponectin concentrations were negatively correlated with insulin concentration and HOMA-IR. In multiple regression analyses, plasma leptin was more positively associated with insulin and proinsulin levels, HOMA-IR and HOMA beta than was adiponectin in boys. This association persisted even after adjusting for body weight, BMI and pubertal status. In conclusion, plasma leptin was more strongly associated with insulin sensitivity and beta-cell function than was adiponectin among children, particularly in boys. PMID- 17109667 TI - Value of intraoperative abdominal cavity culture in appendicectomy: a retrospective study. AB - Appendicectomy is one of the most common surgical emergency procedures. Intraperitoneal culture during appendicectomy is routine practice in some hospitals, while some surgeons advocate abandoning this routine. The aim of our study is to determine the value of intraoperative abdominal cavity culture and its impact on the patient management. Retrospective analysis was performed on 498 patients who underwent appendicectomy over 2.5-year period. The median of the postoperative hospital stay was 2 days, while the median time taken to receive culture results was 3 days. A positive culture was found in 42.6%. Approximately 42.7% of the patients were discharged from the hospital before receiving the culture results. The culture and sensitivity results altered the antibiotic regimen in one patient (0.85%). Intraoperative abdominal cavity culture results were seldom used for clinical management in patients with acute appendicitis. The traditional surgical practice of routinely culturing peritoneal fluid in these patients should be abandoned. PMID- 17109668 TI - Managing enlarged prostate in primary care. AB - Assessment and treatment of benign prostatic hyperplasia, or enlarged prostate, has evolved considerably in recent years; clear evidence has accumulated for the progression of disease over time, the association between disease progression and negative outcomes, and the potential for medical management of this condition. Commensurate with the long-term preventive role of primary care, efforts can and should be made to treat the underlying condition of enlarged prostate as well as to manage the symptoms short-term. This review outlines evaluation of men presenting with lower urinary tract symptoms, examines the challenges for medical treatment and suggests how treatment choice can address these challenges. PMID- 17109669 TI - Dietary sodium intake and asthma: an epidemiological and clinical review. AB - The changes in diet associated with the development of a more affluent lifestyle have been considered one of the environmental factors that may have contributed to the rise in the prevalence of asthma over the past few decades, and dietary sodium has been considered to be a dietary constituent which may be implicated in this phenomenon. The data presented in this review demonstrate that adoption of a low sodium diet for a period of 2-5 weeks may improve lung function and decrease bronchial reactivity in adults with asthma, while sodium loading appears to have a detrimental effect. Similarly, a low sodium diet maintained for 1-2 weeks decreases bronchoconstriction in response to exercise in individuals with asthma. There is no data as to the longer-term effect of a low sodium diet on either the prevalence or severity of asthma or on exercise-induced bronchoconstriction. As a low sodium diet has other beneficial health effects, it can be considered as a therapeutic option for adults with asthma, although it should be considered as an adjunctive intervention to supplement optimal pharmacological management of asthma and not as an alternative. If the relationship between higher sodium intake and increased prevalence and severity of asthma is causal, then there are potential population benefits for asthma as well as cardiovascular disease to be derived from public health measures to reduce sodium consumption. PMID- 17109670 TI - Succinylcholine or rocuronium? A meta-analysis of the effects on intubation conditions. AB - This meta-analysis was undertaken to compare the effects of succinylcholine (SCH) and rocuronium (RCR) on endotracheal intubation (ETI) conditions. Eligible randomised controlled trials (RCTs) were identified via medline and a manual search of references. The frequencies of excellent and unacceptable ETI conditions in subjects receiving SCH 1.0-1.5 mg/kg and RCR 0.6-1.2 mg/kg were determined using standard statistical methods. Inclusion criteria were met by 16 RCTs, representing 1362 subjects. SCH was associated with a 17.7% increase (95% CI = 13-22) in the frequency of excellent ETI conditions and a 5.1% decrease (95% CI = -7.3 to -2.9) in the frequency of unacceptable ETI conditions, when compared with RCR. In the subgroup undergoing true rapid sequence intubation, SCH was associated with a 19.1% increase (95% CI = 13.7-24.5) in the frequency of excellent ETI conditions. SCH appears to be superior to RCR in creating excellent ETI conditions and avoiding unacceptable ETI conditions. PMID- 17109671 TI - Pramlintide in the treatment of diabetes. AB - Diabetes treatment has traditionally focused on correcting insulin deficiency with exogenous insulin and oral agents designed to enhance insulin secretion or insulin sensitivity in peripheral tissues. The more recent view of diabetes as a disease that affects multiple hormones in addition to insulin has led to the development of new therapies more broadly aimed at restoring glucose homeostasis by correcting abnormalities in additional glucoregulatory hormones. Pramlintide, a synthetic analogue of the beta-cell hormone amylin, regulates the appearance of glucose in the circulation following meals through several mechanisms of action: slowing gastric emptying, preventing inappropriate postprandial secretion of glucagon and increasing satiety. Long-term studies have demonstrated that pramlintide improves postprandial glucose fluctuations and A1C while reducing insulin dose and body weight. This combination of benefits associated with pramlintide makes it an attractive new treatment option for patients with diabetes. Clinical Trial Registry Numbers: 137-155 open-label clinical trial: NCT00108004 (Pramlintide long-term, placebo-controlled clinical trials were completed prior to the requirement for NCT registry). PMID- 17109672 TI - Exenatide in type 2 diabetes: treatment effects in clinical studies and animal study data. AB - The therapeutic options for treating type 2 diabetes have been widened by the introduction of exenatide as the first incretin mimetic. Incretins are gut hormones that contribute to the stimulation of insulin secretion after a carbohydrate rich meal. The incretin hormone glucagon-like peptide-1 (GLP-1) not only stimulates insulin secretion under hyperglycaemic conditions, but also suppresses glucagon secretion, slows gastric emptying, induces satiety and improves beta cell function in type 2 diabetes. These beneficial effects have awakened the interest to use GLP-1 for the treatment of type 2 diabetes. Because of its short biological half-life, GLP-1 itself is not practical for type 2 diabetes therapy. Exenatide is a peptide found in the lizard Heloderma suspectum and has a high similarity to GLP-1. Exenatide belongs to the novel class of incretin mimetics because of its incretin-like action. It has a much longer biological half life than GLP-1 and is a GLP-1 receptor agonist that can be used for therapeutic purposes by twice daily injection. Clinical studies and clinical experience with exenatide have shown a significant reduction in HbA1c, fasting- and postprandial glucose and a marked reduction in body weight in type 2 diabetic patients. Animal studies reveal an improvement of beta cell function and an increase in beta cell mass after exenatide treatment. This review gives an overview on exenatide, its pharmacological profile and its role and potential in the therapeutic setting of type 2 diabetes. Furthermore, future developments concerning exenatide application are highlighted. PMID- 17109673 TI - Tigecycline: a new glycylcycline antimicrobial. AB - Tigecycline is a new glycyclcycline antimicrobial recently approved for use in the USA, Europe and elsewhere. While related to the tetracyclines, tigecycline overcomes many of the mechanisms responsible for resistance to this class. It demonstrates favourable in vitro potency against a variety of aerobic and anaerobic Gram-positive and Gram-negative pathogens, including those frequently demonstrating resistance to multiple classes of antimicrobials. This includes methicillin-resistant Staphylococcus aureus, penicillin-resistant S. pneumoniae, vancomycin-resistant enterococci, Acinetobacter baumannii, beta-lactamase producing strains of Haemophilis influenzae and Moraxella catarrhalis, and extended-spectrum beta-lactamase producing strains of Escherichia coli and Klebsiella pneumoniae. In contrast, minimum inhibitory concentrations for Pseudomonas and Proteus spp. are markedly elevated. Tigecycline is administered parenterally twice daily. Randomised, controlled trials have demonstrated that tigecycline is non-inferior to the comparators for the treatment of complicated skin and skin structure infections, as well as complicated intra-abdominal infections. The most frequent and problematic side effect associated with its administration to date has been nausea and/or vomiting. PMID- 17109674 TI - Serial factitious disorder and Munchausen by proxy in pregnancy. AB - Factitious disorder, including Munchausen syndrome, is seldom documented among pregnant patients but can have powerful consequences. We report on a 44-year-old woman who, over a period of two decades, self-induced labour and delivery in five consecutive pregnancies. She precipitated labour by rupturing her own amniotic sac with a fingernail or cervical manipulation, or misappropriating and self administering prostaglandin suppositories from the hospital unit on which she worked as a nurse. Preterm deliveries resulted in fetal demise in one case and in neonatal intensive care treatment for two of the offspring. One of the surviving children has cerebral palsy attributable to the mother's factitious illness behaviour, which raises the spectre of Munchausen by proxy maltreatment. The patient sought attention and care through the ruses, which have never been uncovered by her obstetric and gynaecologic caregivers. Indeed, she underwent an unnecessary hysterectomy because of the illusion of heavy menstrual bleeding. Most recently, the patient has been engaging in surreptitious autophlebotomy to force blood transfusions. PMID- 17109675 TI - Shrinking lungs syndrome, a rare manifestation of systemic lupus erythematosus. AB - In systemic lupus erythematosus (SLE), the respiratory system is frequently compromised. One of its uncommon manifestations is the shrinking lungs syndrome (SLS), characterised by dyspnoea, diaphragmatic elevation and a restrictive pattern in the spirometry. We report two cases affected with this rare entity. They presented with different degrees of respiratory involvement and responses to the therapy. At the same time, clinical, physiopathological, prognostic and therapeutic aspects of this syndrome are reviewed. PMID- 17109676 TI - Continuous Erythropoietin Receptor Activator (C.E.R.A.) administered at extended administration intervals corrects anaemia in patients with chronic kidney disease on dialysis: a randomised, multicentre, multiple-dose, phase II study. AB - This dose-finding, open-label study examined the potential of subcutaneous Continuous Erythropoietin Receptor Activator (C.E.R.A.) to correct anaemia at extended administration intervals in 61 erythropoiesis-stimulating agent-naive patients with chronic kidney disease (CKD) on dialysis. After a 4-week run-in, patients were randomised to C.E.R.A. 0.15, 0.30 and 0.45 microg/kg/week. Within these dose groups, patients were further randomised to once weekly, once every 2 weeks or once every 3 weeks treatment. Mean changes in haemoglobin (Hb) increased with increasing C.E.R.A. dose during a period of 6 weeks where no dose adjustments were permitted. The effect was independent of administration schedule. Erythropoietic responses were sustained until the end of the study (12 weeks) in all groups. In total, 90% of patients in the 0.30 microg/kg/week group and 79% in the 0.45 microg/kg/week group responded to treatment (Hb increase > or =1.0 g/dl), compared with 72% in the 0.15 microg/kg/week group. Faster median response time was associated with increasing dose (51, 38 and 31 days, respectively) and response was unrelated to administration frequency. C.E.R.A. was generally well tolerated. Our results suggest that 0.60 microg/kg twice monthly would be a suitable starting dose of C.E.R.A. for the initiation of anaemia correction in patients with CKD on dialysis. Phase III studies will confirm the feasibility of using C.E.R.A. at extended administration intervals in patients with CKD and anaemia. PMID- 17109677 TI - Rimonabant: endocannabinoid inhibition for the metabolic syndrome. AB - Rimonabant is the first drug to target the endocannabinoid (CB) pathway by inhibiting the actions of anandamide and 2-archidonyl-glycerol on CB1 receptors. This review gives an overview of rimonabant and the CB system and how this system relates to obesity. Rimonabant blocks the central effects of this neurotransmitter pathway involved in obesity and weight control and also blocks the direct effects of CBs on adipocyte and hepatocyte metabolism. Blockade of CB1 receptors leads to a decrease in appetite and also has direct actions in adipose tissue and the liver to improve glucose, fat and cholesterol metabolism so improving insulin resistance, triglycerides and high-density lipoprotein cholesterol (HDL-C) and in some patients, blood pressure. The Rimonabant in Obesity (RIO) trials have shown that rimonabant induces weight loss > 5% in 30 40% of patients and > 10% in 10-20% above both a dietary run-in and long-term hypocaloric management over a 2 year period with a low level of drug-related side effects. Rimonabant therapy is associated with an extra 8-10% increase in HDL-C and a 10-30% reduction in triglycerides and improvements in insulin resistance, glycaemic control in patients with diabetes and also adipokines and cytokines including C-reactive protein over hypocaloric diet therapy. In addition rimonabant abolishes the weight gain associated with smoking cessation and improves the chances of quitting smoking. Thus rimonabant has major effects on both the metabolic syndrome and cardiovascular risk factors thus has the potential to reduce the risks of type 2 diabetes and cardiovascular disease associated with the cardiometabolic phenotype. PMID- 17109678 TI - Chronic hepatitis B: current epidemiology in the Americas and implications for management. AB - Hepatitis B virus (HBV) remains a serious health threat in many parts of the world. Although its prevalence is lower in the Americas than in Asia, Africa and the Middle East, it is responsible for significant morbidity and mortality in North, Central and South America. There is a nonuniform pattern of distribution throughout this region, with HBV prevalence related to geographical, social and cultural factors that predispose certain individuals to infection. This report details the incidence, modes of viral transmission of hepatitis B in the Americas and clinical course of disease in different regions of the Americas. Additionally, the implications for management focusing on issues predominant in high-risk populations are presented. PMID- 17109679 TI - Pretreatment assessment and predictors of hepatitis C virus treatment in US veterans coinfected with HIV and hepatitis C virus. AB - The US Department of Veterans Affairs (VA) cares for many human immunodeficiency virus/hepatitis C virus (HIV/HCV)-coinfected patients. VA treatment recommendations indicate that all HIV/HCV-coinfected patients undergo evaluation for HCV treatment and list pretreatment assessment tests. We compared clinical practice with these recommendations. We identified 377 HIV/HCV-coinfected veterans who began HCV therapy with pegylated interferon and ribavirin and 4135 HIV/HCV-coinfected veterans who did not but were in VA care at the same facilities during the same period. We compared laboratory and clinical characteristics of the two groups and estimated multivariate logistic regression models of receipt of HCV treatment. Overall, patients had high rates of receipt of tests necessary for HCV pretreatment assessment. Patients starting HCV treatment had higher alanine aminotransferase (ALT), lower creatinine, higher CD4 counts and lower HIV viral loads than patients not starting HCV treatment. In the multivariate model, positive predictors of starting HCV treatment included being non-Hispanic whites, having higher ALTs, lower creatinines, higher HCV viral loads, higher CD4 counts, undetectable HIV viral loads and receiving HIV antiretrovirals. A history of chronic mental illness and a history of hard drug use were negative predictors. Most HIV/HCV-coinfected patients received the necessary HCV pretreatment assessments, although rates of screening for hepatitis A and B immunity can be improved. Having well-controlled HIV disease is by far the most important modifiable factor affecting the receipt of HCV treatment. More research is needed to determine if the observed racial differences in starting HCV treatment reflect biological differences, provider behaviour or patient preference. PMID- 17109680 TI - Peginterferon alpha-2b plus ribavirin for treatment of chronic hepatitis C with severe fibrosis: a multicentre randomized controlled trial comparing two doses of peginterferon alpha-2b. AB - We compared sustained virological response (SVR) in chronic hepatitis C patients with severe fibrosis treated with pegylated interferon (Peg-IFN) alpha-2b 1.5 microg/kg/week or 0.75 microg/kg/week in combination with ribavirin 800 mg/day for 48 weeks. This was a multicentre randomized controlled study. SVR was observed in 44.5% (45/101) of patients treated with the standard dose of Peg-IFN and 37.2% (38/102) of patients treated with the low dose (NS). In patients with genotypes 1, 4 and 5, SVR was observed in 25.0% of patients who received the standard dose and 16.9% of patients who received the low dose of Peg-IFN (P = NS). In patients with genotypes 1, 4 and 5 and low viraemia, SVR was obtained in 27.3% of patients treated with the standard dose and 25.8% of patients treated with the low dose (P = NS). In the high-viraemia subgroup, SVR was obtained in 24.0% and 9.1% of patients, respectively. In patients with genotypes 2 and 3, SVR was similar in both groups (73.2%vs 73.0%). Thus, (1) patients with genotypes 2 and 3 and severe fibrosis can be treated with low dose of Peg-IFN and ribavirin, (2) this study suggests that patients with genotypes 1, 4 and 5 and high viraemia could receive a standard dose of Peg-IFN associated with ribavirin for 48 weeks, (3) side effects limit the efficacy of the treatment with standard dose of Peg IFN in patients with genotypes 1, 4 and 5 and low viraemia, (4) more studies are needed for patients with genotype 2 or 3 to define the optimal duration (24 or 48 weeks) in patients with severe fibrosis. PMID- 17109681 TI - Alpha-defensin increase in peripheral blood mononuclear cells from patients with hepatitis C virus chronic infection. AB - The alpha-defensin genes promoter regions contain a putative nuclear factors of activated T cells (NFAT)-binding site and it is known that hepatitis C virus (HCV) core protein activates the interleukin (IL)-2 gene transcription through the NFAT pathway. The aims of this study were to investigate if HCV affects the alpha-defensin expression in peripheral human mononuclear cells (PBMCs) and to evaluate the existence of a correlation between alpha-defensins and liver damage in patients with chronic hepatitis C. Ninety patients with chronic hepatitis C, 30 with chronic hepatitis B and 25 healthy controls were enrolled. Alpha defensins were identified and quantified in PBMCs by mass spectrometry, enzyme linked immunosorbent assay, antibacterial activity and mRNA levels. PBMCs from three patients and controls were stimulated with HCV core protein, hepatitis B virus core antigen and the alpha-defensin mRNAs level was quantified. We found that HCV core protein activates in vitro the alpha-defensin transcription. Alpha defensin levels in patients with chronic hepatitis C (mean +/- SD = 1.103 +/- 0.765 ng/10(6) cells), chronic hepatitis B (0.53 +/- 0.15) and healthy controls (0.217 +/- 0.09) resulted significantly different (P < 0.001). In patients with chronic hepatitis C, the alpha-defensin levels and antibacterial activity correlate with the liver fibrosis. Our data suggest that HCV induces alpha defensin expression. The high linear correlation of alpha-defensin levels with advancing fibrosis makes the measure of these peptides a reliable marker to evaluate fibrosis stage. PMID- 17109682 TI - Impaired health-related quality of life in patients with chronic hepatitis C and persistently normal aminotransferase levels. AB - A significant impact of hepatitis C virus (HCV) infection on health-related quality of life (HRQOL) has been previously described. However, comprehensive data on the quality of life in patients with chronic hepatitis C and persistently normal aminotransferase levels (PNAL) are currently not available. One hundred fifteen patients with chronic hepatitis C (45 with persistently normal aminotransferases and 70 with elevated aminotransferases) and 50 healthy subjects were enrolled. Emotional and psychological states were assessed by Profile of Mood States (POMS) scale and HRQOL was assessed by the 'Everyday Life' questionnaire (EDLQ), a validated questionnaire related to the SF-36 Health Survey. An impairment in HRQOL was observed in patients with chronic hepatitis C showing PNAL compared with healthy subjects with significant differences for the factor scores depression and anger in the POMS scale as well as for the items body, relationship to partner, self-confidence and zest of life in the EDLQ. No differences in any questionnaire were observed between patients with chronic hepatitis C showing PNAL or elevated aminotransferase levels except of a worse mean level for factor score anger in POMS scale in patients with persistently normal aminotransferases. No association of quality of life with severity of liver disease was found. Impairment of HRQOL by chronic infection with HCV is similar in patients with PNAL and those with elevated aminotransferase levels. PMID- 17109683 TI - Quantitative detection of hepatitis E virus RNA and dynamics of viral replication in experimental infection. AB - Hepatitis E virus (HEV) RNA has been detected in the stool and serum of patients with HEV infection and experimentally infected nonhuman primates. However, dynamics of HEV levels in the stool and serum during clinical and subclinical infections have not been determined. A real-time reverse transcription polymerase chain reaction assay, using SYBR Green I in a LightCycler, was developed and optimized to allow quantification of HEV RNA in the stool and serum of both genotype 1 and 2 isolates. The specificity of the assay was confirmed by testing known HEV-RNA-positive and -negative stool and serum specimens and the sensitivity was evaluated using a synthetic HEV RNA standard. Profiles of viraemia and faecal shedding in two chimpanzees inoculated with an isolate of HEV genotype 1 showed the appearance of virus in the stools on day 4 postinoculation (5.65-6.85 log copies/mg) and in the serum on day 7 postinoculation (6.0-6.93 log copies/mL). Peak HEV RNA levels in the stool and serum coincided with peak alanine aminotransferase (ALT) levels observed on day 22 postinoculation in the two chimpanzees. At the time of detection of IgG anti-HEV in serum, viral RNA was no longer detectable in the stool or serum and ALT values had returned to normal levels in both chimpanzees, suggesting the efficacy of the immune response in terminating viral replication. Quantitative evaluation of HEV RNA in humans may allow determining the role of virus levels in the pathogenesis and transmission of HEV. PMID- 17109684 TI - Molecular detection and sequence analysis of a new hepatitis E virus isolate from Pakistan. AB - Sporadic and epidemic acute viral hepatitis E in many developing countries is caused by hepatitis E virus (HEV). The HEV genome has been classified into three major genotypes. However, extensive diversity has been noted among HEV isolates from patients with acute hepatitis in China and Taiwan. Some reports indicated that multiple genotypes of HEV could cocirculate in the same area; even distinct genotypes of HEV could exist in the same patient. Pakistan is a highly endemic area for hepatitis E. So far only two Pakistan HEV isolates Sar-55 (87-Pakistan A) and Abb-2B (88-Pakistan-2B) have been characterized, and the nucleotide sequences of these two HEV isolates show only 90% homology. In this study, a third HEV isolate from Pakistan (87-Pakistan-B) is reported. The sequences of a 438-bp fragment from ORF-2 and a 259-bp fragment from the ORF-1-3 region of this new HEV isolate were obtained and sequenced. The sequence analysis showed that this new HEV isolate was very closely related to the Sar-55 but different from the Abb-2B HEV isolate. These results indicated that the Sar-55 (87-Pakistan-A) genotype is the main endemic HEV strain in the Sargodha area. These data will be useful for HEV epidemiological studies, diagnosis and vaccine development. PMID- 17109685 TI - Hepatitis B virus genotype B is associated with better response to thymosin alpha1 therapy than genotype C. AB - Hepatitis B virus (HBV) genotype has been reported to correlate with response to interferon treatment in several studies. The relationship between HBV genotype and thymosin alpha1 (T-alpha1) treatment is unknown. We retrospectively examine HBV genotypes, precore and core promoter mutations in patients treated with Talpha1 and analyse the correlation between complete response [alanine aminotransferase (ALT) normalization plus seroclearance of HBeAg and HBV-DNA] and HBV genotype. It consisted 98 patients with chronic hepatitis B randomly allocating to three groups: (i) T6 group (n = 32) received a 26-week course of Talpha1 1.6 mg two times a week; (ii) T12 group (n = 34) received the same regimen as T6 group, but Talpha1 therapy extended for 52 weeks; (iii) T0 group (n = 32) served as a control and was followed up for 18 months without specific treatment. Stepwise logistic regression analysis showed that genotype (OR, 3.747; 95% CI, 1.066-13.170; P = 0.039), precore mutation (OR, 6.285; 95% CI, 1.874 21.086; P = 0.003) and Talpha-1 treatment (OR, 12.045; 95% CI, 2.220-65.354; P = 0.004) as independent factors associated with complete response. The complete response of Talpha-1 therapy was higher in patients with genotype B compared to patients with genotype C (52%vs 24%; P = 0.036) and in patients with precore mutation (64%vs 19%; P = 0.002). In conclusion, genotype, presence of precore mutation and Talpha-1 therapy were independent predictors to complete response. Genotype B, compared to genotype C, is associated with a higher response rate to T-alpha1 therapy. PMID- 17109686 TI - Reduction in liver-related hospital admissions and deaths in HIV-infected patients since the year 2002. AB - Since the advent of highly active antiretroviral therapy (HAART), complications of chronic liver disease (CLD) have emerged as one of the leading causes of hospital admission and death among HIV-infected patients with chronic hepatitis B virus (HBV) and/or hepatitis C virus (HCV) infections. The impact of CLD on hospital admissions and deaths in HIV-infected patients attended at one reference HIV hospital in Madrid during the last 9 years was analysed. All clinical charts from January 1996 to December 2004 were retrospectively examined. Demographics, discharge diagnosis, complications during inhospital stay and causes of death were recorded. A total of 2527 hospital admissions in 2008 distinct HIV-infected persons were recorded. Overall, 84% were iv drug users; mean age was 37 years and the mean CD4 count was 224 cells/muL. Both mean age and CD4 count significantly increased during the study period (P < 0.01). Overall, 42% of hospitalized patients were on antiretroviral therapy. Decompensated CLD was the cause of admission and/or developed during hospitalization in 345 patients (14%). Admissions caused by decompensated CLD increased significantly from 9.1% (30/329) in 1996 to 26% (78/294) in 2002. A significant steady decline occurred since then, being 11% (29/253) in the year 2004. Similarly, inhospital liver-related deaths were 9% (5/54) in 1996, peaked to 59% (10/17) in 2001 and declined to 20% (3/15) in the year 2004. Chronic hepatitis C was responsible for admissions and/or deaths in 73.5% of CLD cases. In conclusion, the rate of liver-related hospital admissions and deaths among HIV-infected patients peaked in the year 2002 and has steadily declined since then. A slower progression to liver cirrhosis in patients on HAART, avoidance of hepatotoxic antiretroviral drugs and more frequent use of anti-HCV therapy in HIV/HCV-coinfected patients could account for this benefit. PMID- 17109687 TI - Role of motility in chronic diarrhoea. AB - Patients complaining of 'chronic diarrhoea' usually mean the passage of loose, urgent stools. Chronic diarrhoea is a feature of malabsorption; it may also be seen in the 'dumping syndrome' which follows gastric surgery, small intestinal bacterial overgrowth, bile salt malabsorption and in malabsorption of simple sugars including most commonly lactose, fructose and sorbitol. Excessively rapid entry of chyme into the small or large intestine generates propulsive motor patterns leading to accelerated transit. Inflammation is associated with decreased normal mixing motor patterns but increased propulsive motility including high amplitude propagated contractions (HAPCs). Evidence for abnormal small intestinal motility in the diarrhoea associated with irritable bowel syndrome (IBS) is conflicting and any difference appears small. Increased colonic HAPCs with increased propulsion is seen in IBS with diarrhoea (IBS-D). Stress induced colonic motility is increased in IBS-D with hyper-responsiveness to corticotrophin releasing factor (CRF). Long-lasting increases in mucosal serotonin availability may contribute to the chronic diarrhoea seen in IBS-D and coeliac disease. Treatments for abnormal motility in chronic diarrhoea include those designed to correct specific underlying abnormalities including octreotide, antibiotics, colestyramine, specific food avoidance and anti-inflammatory agents. There are also treatments aimed primarily at altering motility directly including opiates, 5HT3 receptor antagonists and amitriptyline. PMID- 17109688 TI - The gastro-oesophageal common cavity revisited. AB - The manometric common cavity phenomenon has been used as indicator of gastro oesophageal reflux of liquid or gaseous substances. Using combined pH and impedance recording as reference standard the value of a common cavity as indicator of gastro-oesophageal reflux was tested. Ten healthy male subjects underwent combined stationary pressure, pH and impedance recording for 4.5 h. After 1.15 h of recording, a reflux-eliciting meal was consumed. The chi-squared and Kolmogorov-Smirnov tests were used for the statistical analysis. A common cavity was found in 95 (43%) of the 223 reflux events detected by impedance, while seven common cavities were unrelated to a reflux episode. In 54% of the reflux events detected by impedance without a common cavity, a possible common cavity was obscured by either contractile activity or artefacts of various origin. The types of reflux associated with a common cavity (liquid 60%, mixed 31%, gas 9%) and without a common cavity (liquid 59%, mixed 29%, gas 12%) did not differ, or did the acidity of the reflux episodes (with common cavity: acid 67%; without common cavity: acid 58%). The common cavity is a specific but not a sensitive marker of gastro-oesophageal reflux. Furthermore, common cavities are not specific for a particular type of reflux. PMID- 17109689 TI - Measurements of gastric emptying of low- and high-nutrient liquids using 3D ultrasonography and scintigraphy in healthy subjects. AB - Scintigraphy represents the 'gold standard' for the measurement of gastric emptying. Recent studies suggest that three-dimensional (3D) ultrasonography may allow a precise measure of gastric emptying, given the capacity for accurate volume calculations of the stomach. The aim of this study was to compare measurements of gastric emptying of both low- and high-nutrient drinks by 3D ultrasonography with scintigraphy. Ten healthy young subjects (6M, 4F, age 23.5 +/- 1.5 years) were studied on 2 days. Concurrent measurements of gastric emptying by scintigraphy and 3D ultrasonography were performed after ingestion of 500 mL beef soup (12 kcal) or 300 ml dextrose (25% w/v) (314 kcal) labelled with 20 MBq (99m)Tc-sulphur colloid. There was no significant difference between scintigraphic and ultrasonographic 50% emptying times (T50s) (soup: 27.7 +/- 4.8 min vs 23.8. +/- 4.8 min; dextrose: 122.2 +/- 13.3 min vs 131.9 +/- 10.2 min). There was a close correlation between scintigraphic and ultrasonographic T50s for both soup (r = 0.92, P = 0.0005) and dextrose (r = 0.88, P = 0.0007). For the T50s, the limits of agreement were -15.2 min and +8.1 min for the soup (mean difference -3.6 min) and -35.3 min and +47.6 min for dextrose (mean difference +6.2 min). 3D ultrasonography provides a valid measure of gastric emptying of liquid meals in healthy subjects. PMID- 17109690 TI - Pre-experimental stress in patients with irritable bowel syndrome: high cortisol values already before symptom provocation with rectal distensions. AB - Stress is known to affect symptoms of irritable bowel syndrome (IBS) probably by an alteration of visceral sensitivity. We studied the impact of maximal tolerable rectal distensions on cortisol levels in patients with IBS, chronic constipation and controls, and evaluated the effect of the experimental situation per se. In twenty-four IBS patients, eight patients with chronic constipation and 15 controls salivary cortisol was measured before and after repetitive maximal tolerable rectal balloon distensions and at similar times in their usual environment. Rectal sensitivity thresholds were determined. IBS patients but not controls and constipation patients had higher cortisol levels both before and after the experiment compared with similar times on an ordinary day in their usual environment (P = 0.0034 and 0.0002). There was no difference in salivary cortisol level before compared with after rectal distensions. The IBS patients had significantly lower thresholds for first sensation, urge and maximal tolerable distension than controls (P = 0.0247, 0.0001 and <0.0001) and for urge and maximal tolerable distension than patients with constipation (P = 0.006 and 0.013). IBS patients may be more sensitive to expectancy stress than controls and patients with constipation according to salivary cortisol. Rectal distensions were not associated with a further significant increase in cortisol levels. PMID- 17109691 TI - A novel biomagnetic approach to study caecocolonic motility in humans. AB - Motility patterns play a major role in human colonic functions; however, its physiological significance is poorly understood. Several studies have been introducing the Alternating Current Biosusceptometry (ACB) as a valuable tool in gastroenterology and pharmaceutical research. Using gold standard techniques, great effort has been made to validate ACB as a method for measuring gastrointestinal motility in humans and animals. The aim of this study was to evaluate caecocolonic motility and its response to a meal in healthy volunteers. The results showed a dominant frequency of 3.17 +/- 0.13 cycles per minute (mean +/- SD) that remained unchanged even after a standardized meal (P > 0.01). The colonic response to a meal was recorded as a considerable increase in amplitude, reflected by motility index (P < 0.01) and was observed for all the volunteers. The caecocolonic motility could be assessed by the ACB providing new insights into physiological patterns of motility. Moreover, the method is non-invasive, radiation-free, cost-effective and independent of bowel preparation. PMID- 17109692 TI - Effect of the stable thromboxane derivative, carbocyclic thromboxane A2, on membrane potential of rat myenteric neurones in culture. AB - The effects of carbocyclic thromboxane A(2) (cTXA(2); 10(-6) mol L(-1)) on membrane potential and cytosolic Ca(2+) concentration were measured with the whole-cell patch-clamp or the fura-2 method, respectively, at rat myenteric ganglia. cTXA(2) caused a hyperpolarization of myenteric neurones from -19.3 +/- 2.5 to -29.3 +/- 2.3 mV. In addition, the eicosanoid potentiated the carbachol induced depolarization from 4.2 +/- 1.0 mV under control conditions to 11.1 +/- 1.1 mV in the presence of the cTXA(2) (n = 9). The hyperpolarization was abolished by internal application of CsCl (140 mmol L(-1)), a non-selective blocker of K(+) channels, or EGTA (11 mmol L(-1)in the pipette solution), a chelator of intracellular Ca(2+). A similar inhibition was observed in the presence of charybdotoxin (10(-7) mol L(-1)). Fura-2 imaging experiments revealed a cTXA(2)-evoked increase in the intracellular Ca(2+) concentration as indicated by a rise in the fura-2 ratio signal. This response was mediated by a release of Ca(2+) from intracellular stores as sarcoplasmic-endoplasmic reticulum Ca(2+) ATPase blockade with cyclopiazonic acid (5 x 10(-5) mol L(-1)) completely abolished the response to cTXA(2). A similar inhibition was observed after blockade of phospholipase C with U-73122 (10(-5) mol L(-1)). These results suggest an activation of Ca(2+)-activated K(+) channels by cTXA(2) after stimulation of phospholipase C. PMID- 17109693 TI - Fibre-free diet leads to impairment of neuronally mediated muscle contractile response in rat distal colon. AB - Dietary fibre consumption is known to be beneficial to increase stool bulk and frequency. In contrast, it is unclear whether chronic dietary fibre deficiency affects colonic motor functions, especially neuronally mediated muscle contractions. In this study, rats were fed a fibre-free diet or diet containing dietary fibre (cellulose or guar gum) for 27 days. Furthermore, neurogenic and myogenic contractions were evaluated in circular and longitudinal muscle strips of the distal colon. Additionally, the number of enterochromaffin (EC) cells, which play important roles in the initiation of the peristaltic reflex, was also examined by immunohistochemistry for serotonin. Myogenic contractions induced by carbachol or substance P were examined in the presence of tetrodotoxin. Circular muscle was hyposensitive to carbachol, but longitudinal muscle was hypersensitive to substance P in the fibre-free group. Nerve-mediated circular (5-20 Hz) and longitudinal (1-2 Hz) muscle contractions evoked by electrical field stimulation were attenuated in the fibre-free group and the latter response was almost abolished by atropine, suggesting functional changes of cholinergic neurons. EC cell number was decreased in the fibre-free group. In conclusion, changes in neurogenic and myogenic contractions and a decrease in EC cell number observed may affect colonic motility of the fibre-free group. PMID- 17109694 TI - Neural mechanisms of early postinflammatory dysmotility in rat small intestine. AB - Although human postinflammatory dysmotility is known, so far animal studies have primarily investigated changes during inflammation. Here, we focused on postinflammatory changes in rat jejunal myenteric plexus and jejunal motility. Evolution of ethanol/2,4,6-tri-nitrobenzene sulphonic acid (TNBS)-induced inflammation was assessed histologically and by measuring myeloperoxidase activity (MPO). Electromyography and immunohistochemistry were performed 1 week after ethanol/TNBS and also after N(G)-nitro-L-arginine methyl ester (L-NAME) administration. Ethanol/TNBS induced a transient inflammation, with normalization of MPO and histological signs of an early phase of recovery after 1 week. The number of cholinergic neurones was not altered, but myenteric neuronal nitric oxide synthase (nNOS)-immunoreactivity was significantly lower in the early phase of recovery after TNBS compared with water (1.8 +/- 0.2 vs 3.5 +/- 0.2 neurones ganglion(-1), P < 0.001). Interdigestive motility was disrupted with a loss of phase 1 quiescence, an increase of migrating myoelectric complex cycle length, a higher number of non-propagated activity fronts and a decrease of adequately propagated phase 3 s after TNBS. Administration of L-NAME resulted in a similar disruption of interdigestive motility patterns. In the early phase of recovery after ethanol/TNBS-induced jejunal inflammation, a loss of motor inhibition occurs due to a decrease of myenteric nNOS activity. These observations may provide a model for early postinflammatory dysmotility syndromes. PMID- 17109695 TI - Ghrelin augments afferent response to distension in rat isolated jejunum. AB - Ghrelin has been shown to decrease firing of gastric vagal afferents at doses comparable with circulating levels in the fasted state. This raises the possibility that ghrelin may have a hormonal action on other vagal afferent populations. The aim of this study was to determine the effects of ghrelin on jejunal afferent activity; including responses to distension, 2-methyl-5 hydroxytryptamine (2-methyl-5-HT) and cholecystokinin (CCK) in both naive and vagotomized rats. Ghrelin significantly augmented the afferent response to distension. No effect was observed on baseline afferent discharge, or the response to 2-methyl-5-HT and CCK. The effect of ghrelin was more pronounced at lower ramp distending pressures (0-30 mmHg). Similarly, ghrelin augmented the jejunal afferent responses to phasic distension at 10-30 mmHg, but had no effect at higher pressures. Chronic subdiaphragmatic vagotomy and administration of the growth hormone secretagogue receptor (GHS-R) antagonist [D-Lys3]-GHRP-6 prevented the augmentation of the afferent responses to distension indicating ghrelin is acting through the GHS-R on vagal afferent fibres. Ghrelin augments the mechanosensation of jejunal vagal afferents and hence may lead to increased perception of hunger contractions. PMID- 17109696 TI - Evidence for the innervation of the puborectalis muscle by the levator ani nerve. PMID- 17109697 TI - Analysis of magnetic source localization of P300 using the multiple signal classification algorithm. AB - The authors studied the localization of P300 magnetic sources using the multiple signal classification (MUSIC) algorithm. Six healthy subjects (aged 24-34 years old) were investigated with 148-channel whole-head type magnetencephalography using an auditory oddball paradigm in passive mode. The authors also compared six stimulus combinations in order to find the optimal stimulus parameters for P300 magnetic field (P300m) in passive mode. Bilateral MUSIC peaks were located on the mesial temporal, superior temporal and parietal lobes. Interestingly, all MUSIC peaks in these regions emerged earlier in the right hemisphere than in the left hemisphere, suggesting that the right hemisphere has predominance over the left in the processing activity associated with P300m. There were no significant differences among the six stimulus combinations in evoking those P300m sources. The results of the present study suggest that the MUSIC algorithm could be a useful tool for analysis of the time-course of P300m. PMID- 17109698 TI - Stress buffering effects of social support on depressive symptoms in middle age: reciprocity and community mental health. AB - Little is known about the association between depression and the buffering effects of social support in mid-life crisis. The aim of this study is to determine the buffering effects of social support on depression concerning middle aged individuals, while also taking reciprocity and gender differences into careful consideration. A cross-sectional survey of all middle-aged individuals (40-69 years of age) using a large sample (n = 4558) from a community-living population, who resided in Rokunohe town, Aomori prefecture in northern Japan (response rate = 69.8%), was undertaken. This town recently had a lot of suicides. Two-way anova was used to analyze the effects of stressor and social support on the Center for Epidemiologic Studies Depression scale scores. The authors found a stress buffering effect of social support on the depressive symptoms occurring in middle age, however, a significant difference in the stress buffer effect was only observed in male subjects. Moreover, when the authors take reciprocity into account, the effect of the buffer on depression was found not only in males receiving support but in males providing support as well. In conclusion, pertaining to males, social support reduces depressive symptoms under stressful circumstances in middle age, not only when they receive such support but also when they provide it. Therefore, these findings suggest that reciprocal social support is important for males in relation to community mental health. PMID- 17109699 TI - Effects of psychoeducation for depression on help-seeking willingness: biological attribution versus destigmatization. AB - Existing literature shows that the level of biological attribution and stigma of depression influences willingness to seek help. However, no study has used experimental methods to explore the question whether increasing biological attribution and decreasing blameworthy attitude towards depression will enhance willingness to seek help. In so doing, 299 college students were randomly assigned to biological, destigmatization, combined, and control groups. The measures included the Biological Attribution Scale, Psychological Blame Scale, and Help-Seeking Willingness Scale. The data were analyzed by a 2 x 2 ancova (with or without biological attribution education x with or without destigmatization education) on willingness to seek professional help which was assessed 2 weeks later, with adjusting for help-seeking willingness at baseline. Results showed that biological education had a significant main effect to elevate help-seeking willingness, but destigmatization education did not. In addition, no interaction effect existed between the two independent variables. The authors suggested that biological education makes people legitimize depression as a disease entity, so that it would be a practical approach to increase people's motivation to solve their emotional afflictions, especially in societies that emphasize emotional constraints. In contrast, although destigmatization information reduces people's negative appraisals to the depressed individuals, it does not go a step further to increase people's motivation to seek professional help. Further studies are needed to clarify the mechanisms of educational effects. PMID- 17109700 TI - Brain glucose metabolism and temperament in relation to severe somatization. AB - Little is known about the pathophysiology of somatization. The authors' aim was to explore associated factors with somatoform disorders. The authors studied 10 female patients with a diagnosis of somatization disorder or undifferentiated somatoform disorder with no comorbid current Diagnostic and Statistical Manual of Mental Disorders (4th edn) Axis I disorder and 12 healthy female volunteers. The predicting variables were temperament factors of the 240-item Temperament and Character Inventory instrument and regional brain glucose metabolism. Low novelty seeking and high harm avoidance temperament traits and low caudate and low putamen glucose metabolism were statistically significantly associated with severe somatization (P < 0.05). In the present study, severe somatization associates with both altered brain glucose metabolism and temperament factors. No other studies on association of somatization with brain glucose metabolism and temperament have been published. The results are still considered exploratory due to the small number of subjects. PMID- 17109701 TI - Sociopsychological factors relating to suicide prevention in a Japanese rural community: coping behaviors and attitudes toward depression and suicidal ideation. AB - In response to the rise in suicide in Japan since 1998, some suicide prevention measures in local communities have been put into action. However, in the previous suicide prevention measures, sociopsychological factors were not fully taken into consideration. In the present study, the authors surveyed sociopsychological factors relating to suicide and depression (i.e. people's coping behavior and thoughts about depression and suicidal ideation, and their attitudes toward suicide and psychiatric treatment), and their differences in gender and generations. The present study was conducted in a rural area of Japan (Town A), where the suicide rate is much higher than the national average. The authors randomly selected 10% of the residents (i.e. 532 people) aged between 40 and 79 years on the basis of resident registration. Health promotion volunteers in Town A visited these 532 people individually, distributed questionnaires, and asked them to anonymously answer the questionnaire within 4 weeks. Data from 450 residents (193 men, 257 women) were analyzed in the present study. Although there were few gender differences, some significant differences were found between the younger (40-59 years) and older (60-79 years) residents. Generally, the younger were more pessimistic about their mental health than the elderly. It was also noteworthy that about 10% of the people thought that it was natural for them to have suicidal ideation, and about 18% reported that they had experienced suicidal ideation. Some suggestions were given to provide more effective suicide prevention measures. PMID- 17109702 TI - Lymphocyte subsets and viral load in male AIDS patients with major depression: naturalistic study. AB - This study investigated the relationships between lymphocyte subsets and viral load in AIDS patients with and without major depression. During a 7-year period, a total of 60 male hospitalized AIDS patients were recruited in this study. The diagnosis of major depression in patients was made by the same psychiatrist according to the 4th edition of the Diagnostic and Statistical Manual for Mental Disorders criteria. All patients had data for CD4, CD8 and CD4/CD8, and only 42 patients had viral load data. Of 60 AIDS patients, 32 patients had received highly active antiretroviral therapy and received subsequent assessment of lymphocyte cell counts 1 month later. Using ancova with age adjustment, the authors found that patients with major depression (n = 22) had significantly lower CD8 cell counts than patients without major depression (n = 38). However, CD4 cell counts and CD4/CD8 ratios were not significantly different between these two groups. In addition, there was no significant difference in viral load between patients with major depression (n = 16) and patients without major depression (n = 26). Of 32 patients received highly active antiretroviral therapy, 14 patients with an antidepressant (e.g. fluoxetine 20 mg/day) had significantly decreased CD4 and not CD8 cell counts and significantly reduced CD4/CD8 ratios during a period of 1 month. In conclusion, these results suggest that AIDS patients with major depression had significantly lower CD8 cell counts and might have a more severe inflammation/immunity reaction than patients without major depression. PMID- 17109703 TI - Assessment of quality of life with the WHOQOL-BREF in a group of Turkish psychiatric patients compared with diabetic and healthy subjects. AB - Decreased quality of life is often an important cause or consequence of psychiatric illness, and needs to be included in a comprehensive treatment plan. The authors aimed to identify how psychiatric patients characterize the quality of their lives compared to others who are suffering from a chronic physical illness (diabetes) and healthy individuals. A total of 100 psychiatric patients were recruited from Dokuz Eylul University Psychiatry Department outpatient clinic. Of these, 34 had 4(th) edition Diagnostic and Statistical Manual diagnosis of alcohol dependence, 38 had schizophrenia, and 28 had bipolar disorder. A total of 35 patients with diabetes and 49 healthy individuals were also included in the study. The World Health Organization's Quality of Life Questionnaire was used to measure the quality of life. Patients with alcohol dependence, bipolar disorder, and schizophrenia scored lower than healthy subjects on the physical aspects of quality of life. Patients with schizophrenia had lower scores in the psychological domain compared to patients with bipolar disorder, patients with diabetes, and healthy subjects. In the social relationship domain, patients with schizophrenia and alcohol dependence scored lower compared to healthy subjects. Patients with schizophrenia were worse with respect to social relationships than bipolar patients and diabetics. World Health Organization's Quality of Life Questionnaire is useful for evaluating the needs and targets for interventions in psychiatric patients. PMID- 17109704 TI - Depression and somatic symptoms scale: a new scale with both depression and somatic symptoms emphasized. AB - The authors' preliminary study selected 22 items for Depression and Somatic Symptoms Scale (DSSS), including depression subscale (DS) and somatic subscale (SS). The aim of the study was to test reliability and validity of the DSSS. The study enrolled 135 consecutive outpatients (34 male and 101 female) experiencing a major depressive episode (the MDE group), 95 of whom (25 male and 70 female) accepted 1 month of treatment (the treatment group). Diagnosis was confirmed by using the Structured Clinical Interview for 4(th) edition with text revision Diagnostic and Statistical Manual Axis I Disorders. The DSSS and Hamilton Depression Rating Scale (HAMD) were given and evaluated. Cronbach's alpha was used to assess internal consistency. The correlation between the improvement percentage (IP) for the HAMD and the IP for the DSSS was calculated for the treatment group. Factor analysis was performed by using the principal-axis factoring method with promax rotation. Cronbach's alpha values of the DSSS and its subscales ranged from 0.73 to 0.94. Pearson correlation coefficients for the relationship between the DSSS and HAMD ranged from 0.63 to 0.86. In the treatment group, DSSS and HAMD scores were significantly decreased after treatment and the IP for the HAMD and the DSSS were similar and correlated (correlation coefficient = 0.78). The results of the factor analysis demonstrated that most of the items in DS and SS appropriately loaded in Depression and Somatic factors, respectively. The discriminative ability of the DSSS for anxiety comorbidities was not inferior to that of the HAMD. Therefore, the DSSS is reliable and sensitive to the treatment and has acceptable convergent, factorial, and distinct groups validities. Because it assesses both depression and somatic symptoms, DSSS may overcome the deficiency of other depression scales with few somatic items. PMID- 17109705 TI - Do staffing levels determine outcome in psychiatric inpatient care? - Factors related to the ratio of period hospitalized in Japan. AB - The aim of this study was to determine the factors related to outcome, as measured by the cumulative number of days hospitalized, of a cohort group of inpatients with a diagnosis of schizophrenia followed up for 6 years. Comprehensive data were obtained from 18 psychiatric hospitals from two surveys conducted in 1993 and 1999. Outcome was evaluated by calculating the cumulative number of days between the two surveys that the cohort group was hospitalized. Tree-based models analysis was used to explore the factors related to outcome. Patient characteristics at the time of the first survey that were related to outcome were the number of days hospitalized, cumulative number of years hospitalized, age, Global Assessment of Functioning score and relationship to the main visitor. The number of beds in each hospital and changes in their number were also related. Increasing staffing levels is not likely to impact on deinstitutionalization since their levels were not related to outcome. PMID- 17109706 TI - Effect of cognitive training focusing on organizational strategies in patients with obsessive-compulsive disorder. AB - The purpose of the present paper was to develop a cognitive training program for patients with obsessive-compulsive disorder (OCD) and evaluate its effectiveness. Nine 60-min sessions focusing on the improvement of organizational strategies were given to 15 patients with OCD over a period of 5 weeks. The control group consisted of 15 age- and sex-matched patients also with OCD. The Rey-Osterrieth Complex Figure Test and Korean-California Verbal Learning Test were administered before and after cognitive training. Clinical symptoms were assessed with the Yale-Brown Obsessive-Compulsive Scale. The memory function in the treatment group improved and their clinical symptoms were alleviated after training, compared to those of the control group. Cognitive training of OCD patients not only improved their memory function, but also alleviated their clinical symptoms. Therefore, cognitive training, focusing on the improvement of organizational strategies, could be an effective treatment modality for patients with OCD. PMID- 17109707 TI - Countertransference to psychiatric patients in a clinical setting: development of the Feeling Checklist-Japanese version. AB - Countertransference is an important dimension of the therapeutic alliance between care providers and patients. The Feeling Checklist (FC) is a self-report questionnaire for the assessment of countertransference by hospital staff toward patients. The FC was translated from English into Japanese and its factor structure, reliability, and validity in the Japanese version (FC-J) were examined. A total of 281 Japanese psychiatric nurses were tested with the FC-J. All nurses were primarily involved in provision of psychiatric care. Principal component factor analysis with varimax rotation was performed to identify the potential components of the FC-J. In a factor analysis of the FC-J, seven factors were extracted. The five subscales that were determined and labeled included Reject, Distance, Helpfulness, Closeness, and Involvement, which collectively accounted for 56.0% of the variance. Cronbach's alpha, a measure of internal consistency, for individual subscales was 0.833 for Reject, 0.763 for Distance, 0.768 for Helpfulness, 0.617 for Closeness, and 0.663 for Involvement. Notably, there was a significant correlation between the FC-J and the Nurse Attitude Scale (P < 0.0001). Moreover, one-way anova was performed with each FC-J subscale to examine differences among psychiatric diagnoses in the study sample. A significant difference was found for Involvement (P < 0.001), with the total score on Involvement being the highest in the personality disorder group. These results are considered to verify the reliability and validity of the FC-J as a scale to measure countertransference among Japanese care providers. The use of this scale allows individual care providers to recognize and be cognizant of their own countertransference objectively and thereby contributes to improve the relationship between patients and care providers. PMID- 17109708 TI - Causative factors as cues for addressing the rapid increase in suicide in Mie Prefecture, Japan: comparison of trends between 1996-2002 and 1989-1995. AB - The number of suicides in Japan has increased from approximately 22 000 per year from 1988 to 1997 to >30 000 per year since 1998. Likewise, the number of suicides has been increasing in Mie Prefecture. The purpose of the present study was to examine the incidence of and circumstances surrounding all suicide cases during 1996-2002 in Mie Prefecture and to compare the data with those from 1989 to 1995. In Mie Prefecture, the age-specific suicide rate during the second 7 year period included marked increases among men aged 50-59 and 60-69 years. Among women, the age-specific suicide rate increased with age during both 7-year periods. During the second period, psychiatric disorders as causative factors increased in all generations. They were especially important for women of the younger generation, whereas economic problems were the most common causative factor among men aged 40-64. Physical illness as a causative factor in suicide was high among the elderly, but among the other age groups this factor trailed behind economic difficulties for men and psychiatric disorders for women. To prevent suicide, social cooperation as well as a plenitude of visiting nurses and psychiatric care is required, and early detection and treatment are also important. PMID- 17109709 TI - Positive correlation between anxiety severity and plasma levels of dehydroepiandrosterone sulfate in medication-free patients experiencing a major episode of depression. AB - Although numerous studies have identified a correlation between dehydroepiandrosterone sulfate (DHEAS) levels and anxiety or depression, those findings remain controversial. The purpose of the present study was to determine whether a correlation exists between depression severity and anxiety severity and serum DHEAS concentrations in medication-free patients experiencing a major depressive episode. Twenty-eight medication-free major depressive outpatients (Hamilton Rating Scale for Depression 17 [HAM-D 17] score >or=17) were enrolled consecutively. Plasma DHEAS levels of all subjects were measured. Blood from subjects was drawn at 0900-1100 h Depression severity was assessed with the HAM-D 17 and the Hospital Anxiety and Depression Scale (HADS) depression subscale. Anxiety was assessed using the HADS anxiety subscale. Serum concentrations of DHEAS were measured immediately following the HAM-D 17 and HADS assessments. A significant, positive correlation was identified between HADS anxiety subscale total score and morning serum DHEAS concentration (P = 0.013) after controlling for age, gender and body mass index (BMI). No statistically significant correlations were found between depression ratings and morning serum DHEAS concentrations. This preliminary study provides pilot data indicating that morning serum DHEAS concentrations were positively correlated with HADS anxiety subscale score (anxiety severity) after controlling for age, gender and BMI in medication-free outpatients experiencing a major depressive episode. It is not known if morning serum DHEAS levels would show similar or dissimilar changes in non-depressed subjects. The present result needs subsequent replication. PMID- 17109710 TI - Effects of switching from conventional antipsychotics to risperidone in Japanese patients with chronic schizophrenia. AB - Atypical antipsychotics are rapidly evolving to become the standard pharmacotherapy in schizophrenia; however, the trend of switching to such drugs is not necessarily progressing quickly in East Asia. This might be due to the scarcity of evidence for the efficacy of switching from conventional to atypical antipsychotics, which prompted the authors to examine effects of switching from conventional antipsychotics to an atypical drug, risperidone, in Japanese patients. Fifty patients with chronic schizophrenia completed the study in which combination therapy with other antipsychotics was allowed if monotherapy with risperidone was not tolerated. Symptoms were assessed with the brief psychiatric rating scale (BPRS). Switching to monotherapy was achieved in 34 patients (68%). The number of antipsychotics prescribed to each patient was reduced (from 2.1 to 1.4 drugs; P < 0.001) and the use of antiparkinsonian drugs decreased (P < 0.001). The mean BPRS score was also reduced 6 months after initiation of the switch (P < 0.001). Failure in switching to monotherapy was associated with higher dosage of antipsychotics at baseline. Switching from conventional antipsychotics to risperidone reduced schizophrenia symptoms, antiparkinsonian medication, and polypharmacy. However, a portion of patients, particularly those who receive an excessive dosage of antipsychotics, might not tolerate such switching. PMID- 17109711 TI - Wilson's disease associated with delusional disorder. AB - Wilson's disease (WD), or hepatolenticular degeneration, is a genetic neurodegenerative disorder of copper metabolism. It is an uncommon medical condition that produces psychiatric symptoms during the early phase in approximately 50% of patients. Reported herein is a case of WD in a young man presenting persistent delusional disorder of organic etiology, which resolved entirely after 4 months of combined pharmacotherapy. The present case demonstrates the importance of considering the occurrence of psychotic symptoms in WD patients given that psychiatric manifestations in WD are known to be uncommon as well as inhomogeneous. It also supports the hypothesis that psychopathologic features in WD have an organic foundation. PMID- 17109712 TI - Reversible dementia with psychosis: Hashimoto's encephalopathy. AB - A case of presumed Hashimoto's encephalopathy (HE) is presented. The presentation included memory loss, delusions, functional decline and culminated in a generalized seizure. Anti-thyroid antibodies were detected and symptoms resolved with prednisolone. Patients with HE may present with prominent neuropsychiatric symptoms, attract psychiatric diagnoses and present to psychiatric services. Primarily a diagnosis of exclusion, HE should be considered in cases of encephalopathy in which standard investigations are negative. PMID- 17109713 TI - Schizotypy, attention deficit hyperactivity disorder, and dopamine genes. AB - Previous research has suggested that there may be overlap between schizophrenia and attention-deficit hyperactivity disorder (ADHD). The relationship between schizotypal personality traits, ADHD features and polymorphisms was evaluated in dopamine-related genes. Thirty-one healthy, Caucasian men completed the Rust Inventory of Schizotypal Cognitions (RISC) and the ADHD Self-Report Scale (ASRS). Catechol-O-methyltransferase (COMT) Val158Met, dopamine receptors of the D3 type (DRD3) Ser9Gly, DRD4 variable number of tandem repeats (VNTR), and SLC6A3 VNTR polymorphisms were analyzed. RISC score was correlated with ASRS score (r = 0.54, P = 0.003). COMT Met homozygotes had higher ASRS scores than Val homozygotes (P = 0.005). These findings are consistent with evidence of overlap between schizophrenia and ADHD and support an involvement of COMT genotype in ADHD features. PMID- 17109714 TI - Benzodiazepines in catatonia associated with systemic lupus erythematosus. AB - Neuropsychiatric disturbances are found in 50-70% of systemic lupus erythematosus (SLE) patients. However, there are rare cases of catatonia being described in SLE. Some studies have shown the effectiveness of high-dose steroid, plasma exchange and electroconvulsive therapy (ECT) in lupus catatonia. Herein are described two SLE patients with catatonia who had good response to i.v. diazepam (i.e. relief of catatonia symptoms). Patient 1, with mild cortical atrophy, had great improvement in catatonia symptoms on i.v. diazepam 150 mg during a period of 5 days. Patient 2, without cortical atrophy, had quick response to i.v. diazepam 10-20 mg. Both patients had no recurrence during 6-month follow up. In conclusion, benzodiazepines may play an important role in the treatment of catatonia associated with SLE if patients refuse ECT treatment. PMID- 17109715 TI - Pisa syndrome-like peculiar posture occurred while running was successfully improved with risperidone. PMID- 17109716 TI - Werner syndrome with psychosis. PMID- 17109717 TI - Mania as a first presentation in mitochondrial myopathy. PMID- 17109718 TI - Transient administration of triclofos sodium is helpful in minimizing anticonvulsants under polypharmacy. PMID- 17109719 TI - Prescription pattern of antipsychotic drugs for schizophrenic inpatients in Japan: research on East Asia Psychotropic Prescription Pattern-Antipsychotics study. PMID- 17109720 TI - Video-assisted cognitive behavioral therapy for anorexia nervosa. PMID- 17109721 TI - Significant correlation of yearly suicide rates with the rate of unemployment among men results in a rapid increase of suicide in Mie Prefecture, Japan. PMID- 17109722 TI - Eye of the needle. PMID- 17109723 TI - Randomized controlled trial of tacrolimus versus microemulsified cyclosporin (TMC) in liver transplantation: poststudy surveillance to 3 years. AB - The 1-year results of the tacrolimus versus microemulsified cyclosporin (TMC) study found a benefit with tacrolimus immunosuppression after primary liver transplants in adults with respect to freedom from graft loss and immunological failure. The integrity of the randomization process was preserved for a further 2 years for poststudy surveillance. The data after 3 years confirms the significant difference between tacrolimus and cyclosporin with tacrolimus less likely to meet the composite primary endpoint (log rank p = 0.01; relative risk 0.75; 95% CI 0.60-0.95; p = 0.016). However, freedom from death or retransplantation no longer achieves statistical significance (relative risk 0.79; 95% CI 0.62-1.02; p = 0.065). A total of 62.1% of patients randomized to tacrolimus were alive at 3 years with their original graft and still on their allocated study medication, as compared with only 41.6% in the cyclosporin limb (p < 0.001). No difference was detected between tacrolimus and cyclosporin in hepatitis-C-positive patients with the available data. The TMC study confirms after 3 years of follow-up the benefits of tacrolimus-based immunosuppression over cyclosporin using C(0) monitoring. PMID- 17109725 TI - Immunoadsorption in severe C4d-positive acute kidney allograft rejection: a randomized controlled trial. AB - Antibody-mediated rejection (AMR) frequently causes refractory graft dysfunction. This randomized controlled trial was designed to evaluate whether immunoadsorption (IA) is effective in the treatment of severe C4d-positive AMR. Ten out of 756 kidney allograft recipients were included. Patients were randomly assigned to IA with protein A (N = 5) or no such treatment (N = 5) with the option of IA rescue after 3 weeks. Enrolled recipients were subjected to tacrolimus conversion and, if indicated, 'anti-cellular' treatment. All IA treated patients responded to treatment. One death unrelated to IA occurred after successful reversal of rejection. Four control subjects remained dialysis dependent. With the exception of one patient who developed graft necrosis, non responders were subjected to rescue IA, however, without success. Because of a high graft loss rate in the control group the study was terminated after a first interim analysis. Even though limited by small patient numbers, this trial suggests efficiency of IA in reversing severe AMR. PMID- 17109726 TI - American Society of Transplantation executive summary on pediatric lung transplantation. AB - Lung transplantation in children poses distinctly different challenges from those seen in the adult population. This consensus statement reviews the experience in the field of pediatric lung transplantation and highlights areas that deserve further investigation. PMID- 17109727 TI - Prevalence and risk factors of non-adherence with immunosuppressive medication in kidney transplant patients. AB - Non-adherence with immunosuppressive regimen is a major risk factor for poor outcome after kidney transplantation. Identifying patients at risk for non adherence requires understanding the risk factors for non-adherence. This prospective study included a convenience sample of 249 adult kidney transplant patients >1 year post-transplant. Non-adherence was monitored electronically using MEMS(R). Selected socio-economic, therapy-, patient-, condition- and healthcare team-related risk factors for non-adherence were assessed. Period prevalences were expressed as the percent of prescribed doses taken (taking adherence), the percent of correctly dosed days (dosing adherence), the percentage of inter-dose intervals not exceeding 25% of the prescribed interval (timing adherence), and the number of drug holidays per 100 days (no intake for > 48 h if once daily or for > 24 h if twice daily intake). Testing occurred by simple mixed logistic regression analysis. Factors significant after correction for multiple testing were entered into a multiple logistic regression model. Mean taking, dosing, timing adherence, and drug holidays were 98%, 96%, 93%, and 1.1 days, respectively. Non-adherence was associated with lower self-efficacy, higher self-reported non-adherence, no pillbox usage, and male gender. Adherence declined between Monday and Sunday. This study provides a framework for identifying patients at risk for non-adherence and for developing adherence enhancing interventions. PMID- 17109728 TI - Growing pains from the islet cell transplant world. PMID- 17109730 TI - Prevalence and estimated incidence of blood-borne viral pathogen infection in organ and tissue donors from northern Alberta. AB - To determine the potential safety benefit of introducing nucleic acid testing (NAT) in tissue and organ donors, the risk of virus transmission was examined in a Canadian population. Anonymous data on Northern Alberta tissue and organ donors from 1998 to 2004 were used to determine the seroprevalence and estimate the seroincidence and residual risk of HIV, HBV, HCV and HTLV infection. Of the 3372 donors identified, 71.1% were surgical bone, 13.2% were living organ and 15.6% were deceased organ/tissue donors. Seroprevalence was: HIV 0.00%, HBV 0.09%, HCV 0.48% and HTLV 0.03%. Incidence (/100,000 p-yrs) and residual risks (/100,000 donors) could only be estimated for HBV (24.2 and 3.9) and HCV (11.2 and 2.2). Risk estimates were higher for deceased donors than surgical bone donors. HCV had the highest prevalence and HBV had the highest estimated incidence. HIV and HTLV risks were extremely low precluding accurate quantification. In this region of low overall viral prevalence, HCV NAT would be most effective in deceased organ donors. In surgical bone donors the cost of implementing NAT is high without significant added safety benefit. PMID- 17109731 TI - Missed opportunities: the institute of medicine report: organ donation: opportunities for action. PMID- 17109732 TI - Recurrent nephrotic syndrome in homozygous truncating NPHS2 mutation is not due to anti-podocin antibodies. AB - Mutations in NPHS2 are a common cause of focal segmental glomerulosclerosis (FSGS). It was initially assumed that FSGS caused by a genetically defective protein in the native kidney would not recur after transplantation; however, description of three patients with NPHS2 missense mutations challenged the validity of this assumption. A possible mechanism of recurrence in cases with stop-codon mutations is the formation of auto-antibodies against the truncated protein. In this case report, we describe a 9-year-old girl with the R138X NPHS2 mutation who presented with recurrent nephrotic syndrome 4 years after renal transplantation from a deceased donor, and was treated with plasmapheresis with a partial response. Renal histology did not demonstrate glomerular immunoglobulin deposition and an extensive search for anti-podocin antibodies based on indirect Western blot with recombinant podocin, was negative, as was the test for glomerular permeability factor (Palb). Taken together these findings confirm the possibility of post transplantation nephrotic syndrome in patients with NPHS2 mutations. Lack of immunoglobulin deposition, absence of circulating anti-podocin antibodies, and normal Palb suggest that other, unknown pathogenetic mechanisms are implicated. PMID- 17109733 TI - Following anti-CD25 treatment, a functional CD4+CD25+ regulatory T-cell pool is present in renal transplant recipients. AB - Daclizumab, a humanized antibody directed against the alpha-chain of the interleukin-2 receptor (CD25), has shown efficacy in the prevention of acute rejection following organ transplantation. However, anti-CD25 therapy can be expected to affect not only alloreactive effector T cells, but also CD4(+)CD25(+) regulatory T (Treg) cells that are shown to play an important role in the induction of transplantation tolerance. Therefore, the size and function of the Treg pool in human renal allograft recipients after single-dose daclizumab administration was investigated in this study. Approximately 8 weeks after administration, daclizumab was cleared from the circulation and the Treg population then present appeared not different from that observed before transplantation. Functional analysis revealed that the Treg possessed a normal capacity to suppress mixed lymphocyte reactions in vitro. These data indicate that after daclizumab therapy a Treg population, normal in number and function, is present in the peripheral blood of renal transplant recipients. PMID- 17109734 TI - Prolonged function of macrophage, von Willebrand factor-deficient porcine pulmonary xenografts. AB - Porcine von Willebrand factor (vWF) activates human and primate platelets. Having determined the importance of pulmonary intravascular macrophages (PIMs) in pulmonary xenotransplantation, we evaluated whether, in the absence of PIMs, vWF might play a role in pulmonary xenograft dysfunction. Utilizing a left single lung transplant model, baboons depleted of anti-alphaGal antibodies received lungs from either vWF-deficient (n = 2); MCP-expressing (n = 5); MCP PIM-depleted (n = 5); or vWF-deficient PIM-depleted swine (n = 3). Two out of three of the PIM depleted, pvWF deficient grafts survived longer than any previously reported pulmonary xenografts, including PIM-depleted xenografts expressing human complement regulatory proteins. Depletion of PIM's from vWF-deficient lungs, like depletion of PIM's from hMCP lungs, resulted in abrogation of the coagulopathy associated with pulmonary xenotransplantation. Thus, in terms of pulmonary graft survival, control of adverse reactions involving pvWF appears to be equally or even more important than is complement regulation using hMCP expression. However, based on the rapid failure of PIM-sufficient, pvWF-deficient pulmonary xenografts, pVWF-deficient pulmonary xenografts appear to be particularly sensitive to macrophage-mediated damage. These data provide initial evidence that vWF plays a role in the 'delayed' (24 h) dysfunction observed in pulmonary xenotransplantation using PIM depleted hMCP organs. PMID- 17109735 TI - Acute renal disease, as defined by the RIFLE criteria, post-liver transplantation. AB - Acute renal failure (ARF) can complicate up to 60% of orthotopic liver transplants (OLT). The RIFLE criteria were developed to provide a consensus definition for acute renal disease in critically ill patients. Using the RIFLE criteria, we aimed to determine the incidence and risk factors for ARF and acute renal injury (ARI), and to evaluate the link with the outcomes, patient survival and length of hospital stay. Three hundred patients, who received 359 OLTs, were retrospectively analyzed. ARI and ARF occurred post 11.1 and 25.7% of OLTs, respectively. By multivariate analysis, ARI was associated with pre-OLT hypertension and alcoholic liver disease and ARF with higher pre-OLT creatinine, inotrope and aminoglycoside use. ARF, but not ARI, had an impact on 30-day and 1 year patient survival and longer length of hospital stay. ARI and ARF, as defined by the RIFLE criteria, are common complications of OLT, with distinct risk factors and ARF has serious clinical consequences. The development of a consensus definition is a welcome advance, however these criteria do need to be validated in large studies in a wide variety of patient populations. PMID- 17109736 TI - Diagnosis of early coagulation abnormalities in trauma patients by rotation thrombelastography. AB - BACKGROUND: Reagent-supported thromboelastometry with the rotation thrombelastography (e.g. ROTEM) is a whole blood assay that evaluates the visco elastic properties during blood clot formation and clot lysis. A hemostatic monitor capable of rapid and accurate detection of clinical coagulopathy within the resuscitation room could improve management of bleeding after trauma. OBJECTIVES: The goals of this study were to establish whether ROTEM correlated with standard coagulation parameters to rapidly detect bleeding disorders and whether it can help to guide transfusion. METHODS: Ninety trauma patients were included in the study. At admission, standard coagulation assays were performed and ROTEM parameters such as clot formation time (CFT) and clot amplitude (CA) were obtained at 15 min (CA(15)) with two activated tests (INTEM, EXTEM) and at 10 min (CA(10)) with a test analyzing specifically the fibrin component of coagulation (FIBTEM). RESULTS: Trauma induced significant modifications of coagulation as assessed by standard assays and ROTEM. A significant correlation was found between prothrombin time (PT) and CA(15)-EXTEM (r = 0.66, P < 0.0001), between activated partial thromboplastin time and CFT-INTEM (r = 0.91, P < 0.0001), between fibrinogen level and CA(10)-FIBTEM (r = 0.85, P < 0.0001), and between platelet count and CA(15)-INTEM (r = 0.57, P < 0.0001). A cutoff value of CA(15)-EXTEM at 32 mm and CA(10)-FIBTEM at 5 mm presented a good sensitivity (87% and 91%) and specificity (100% and 85%) to detect a PT > 1.5 of control value and a fibrinogen less than 1 g L(-1), respectively. CONCLUSIONS: ROTEM is a point-of care device that rapidly detects systemic changes of in vivo coagulation in trauma patients, and it might be a helpful device in guiding transfusion. PMID- 17109737 TI - Successful prophylaxis against intracranial hemorrhage using weekly administration of activated recombinant factor VII in a newborn with severe factor VII deficiency. PMID- 17109738 TI - Announcing a TAFIa mutant with a 180-fold increased half-life and concomitantly a strongly increased antifibrinolytic potential. PMID- 17109739 TI - Sleep characteristics of persons with chronic fatigue syndrome and non-fatigued controls: results from a population-based study. AB - BACKGROUND: The etiology and pathophysiology of chronic fatigue syndrome (CFS) remain inchoate. Attempts to elucidate the pathophysiology must consider sleep physiology, as unrefreshing sleep is the most commonly reported of the 8 case defining symptoms of CFS. Although published studies have consistently reported inefficient sleep and documented a variable occurrence of previously undiagnosed primary sleep disorders, they have not identified characteristic disturbances in sleep architecture or a distinctive pattern of polysomnographic abnormalities associated with CFS. METHODS: This study recruited CFS cases and non-fatigued controls from a population based study of CFS in Wichita, Kansas. Participants spent two nights in the research unit of a local hospital and underwent overnight polysomnographic and daytime multiple sleep latency testing in order to characterize sleep architecture. RESULTS: Approximately 18% of persons with CFS and 7% of asymptomatic controls were diagnosed with severe primary sleep disorders and were excluded from further analysis. These rates were not significantly different. Persons with CFS had a significantly higher mean frequency of obstructive apnea per hour (p = .003); however, the difference was not clinically meaningful. Other characteristics of sleep architecture did not differ between persons with CFS and controls. CONCLUSION: Although disordered breathing during sleep may be associated with CFS, this study generally did not provide evidence that altered sleep architecture is a critical factor in CFS. Future studies should further scrutinize the relationship between subjective sleep quality relative to objective polysomnographic measures. PMID- 17109740 TI - Incongruence between genetic and morphological diversity in Microcebus griseorufus of Beza Mahafaly. AB - BACKGROUND: The past decade has seen a remarkable increase in the number of recognized mouse lemur species (genus Microcebus). As recently as 1994, only two species of mouse lemur were recognized according to the rules of zoological nomenclature. That number has now climbed to as many as fifteen proposed species. Indeed, increases in recognized species diversity have also characterized other nocturnal primates--galagos, sportive lemurs, and tarsiers. Presumably, the movement relates more to a previous lack of information than it does to any recent proclivity for taxonomic splitting. Due to their nocturnal habits, one can hypothesize that mouse lemurs will show only minimal variation in pelage coloration as such variation should be inconsequential for the purposes of mate and/or species recognition. Even so, current species descriptions for nocturnal strepsirrhines place a good deal of emphasis on relatively fine distinctions in pelage coloration. RESULTS: Here, we report results from a multi-year study of mouse lemur populations from Beza Mahafaly in southern Madagascar. On the basis of morphological and pelage variation, we initially hypothesized the presence of up to three species of mouse lemurs occurring sympatrically at this locality, one of which appeared to be undescribed. Genetic analysis reveals definitively, however, that all three color morphs belong to a single recognized species, Microcebus griseorufus. Indeed, in some cases, the three color morphs can be characterized by identical mitochondrial haplotypes. CONCLUSION: Given these results, we conclude that investigators should always proceed with caution when using a single data source to identify novel species. A synthetic approach that combines morphological, genetic, geographic, and ecological data is most likely to reveal the true nature of species diversity. PMID- 17109742 TI - Morphological characterisation of portal myofibroblasts and hepatic stellate cells in the normal dog liver. AB - BACKGROUND: Hepatic fibrosis is a common outcome of hepatic injury in both man and dog. Activated fibroblasts which develop myofibroblastic characteristics play an essential role in hepatic fibrogenesis, and are comprised of three subpopulations: 1) portal or septal myofibroblasts, 2) interface myofibroblasts and 3) the perisinusoidally located hepatic stellate cells (HSC). The present study was performed to investigate the immunohistochemical characteristics of canine portal myofibroblasts (MF) and HSC in the normal unaffected liver as a basis for further studies on fibrogenesis in canine liver disease. RESULTS: In the formalin-fixed and paraffin embedded normal canine liver vimentin showed staining of hepatic fibroblasts, probably including MF in portal areas and around hepatic veins; however, HSC were in general negative. Desmin proved to react with both portal MF and HSC. A unique feature of these HSC was the positive immunostaining for alpha-smooth muscle actin (alpha-SMA) and muscle-specific actin clone HHF35 (HHF35), also portal MF stained positive with these antibodies. Synaptophysin and glial fibrillary acidic protein (GFAP) were consistently negative in the normal canine liver. In a frozen chronic hepatitis case (with expected activated hepatic MF and HSC), HSC were negative to synaptophysin, GFAP and NCAM. Transmission electron microscopy (TEM) immunogold labelling for alpha SMA and HHF35 recognized the positive cells as HSC situated in the space of Disse. CONCLUSION: In the normal formalin-fixed and paraffin embedded canine liver hepatic portal MF and HSC can be identified by alpha-SMA, HHF35 and to a lesser extent desmin immunostaining. These antibodies can thus be used in further studies on hepatic fibrosis. Synaptophysin, GFAP and NCAM do not seem suitable for marking of canine HSC. The positivity of HSC for alpha-SMA and HHF35 in the normal canine liver may eventually reflect a more active regulation of hepatic sinusoidal flow by these HSC compared to other species. PMID- 17109743 TI - Readability of pediatric health materials for preventive dental care. AB - BACKGROUND: This study examined the content and general readability of pediatric oral health education materials for parents of young children. METHODS: Twenty seven pediatric oral health pamphlets or brochures from commercial, government, industry, and private nonprofit sources were analyzed for general readability ("usability") according to several parameters: readability, (Flesch-Kincaid grade level, Flesch Reading Ease, and SMOG grade level); thoroughness, (inclusion of topics important to young childrens' oral health); textual framework (frequency of complex phrases, use of pictures, diagrams, and bulleted text within materials); and terminology (frequency of difficult words and dental jargon). RESULTS: Readability of the written texts ranged from 2nd to 9th grade. The average Flesch-Kincaid grade level for government publications was equivalent to a grade 4 reading level (4.73, range, 2.4-6.6); F-K grade levels for commercial publications averaged 8.1 (range, 6.9-8.9); and industry published materials read at an average Flesch-Kincaid grade level of 7.4 (range, 4.7-9.3). SMOG readability analysis, based on a count of polysyllabic words, consistently rated materials 2 to 3 grade levels higher than did the Flesch-Kincaid analysis. Government sources were significantly lower compared to commercial and industry sources for Flesch-Kincaid grade level and SMOG readability analysis. Content analysis found materials from commercial and industry sources more complex than government-sponsored publications, whereas commercial sources were more thorough in coverage of pediatric oral health topics. Different materials frequently contained conflicting information. CONCLUSION: Pediatric oral health care materials are readily available, yet their quality and readability vary widely. In general, government publications are more readable than their commercial and industry counterparts. The criteria for usability and results of the analyses presented in this article can be used by consumers of dental educational materials to ensure that their choices are well-suited to their specific patient population. PMID- 17109744 TI - Bernard-Soulier syndrome (hemorrhagiparous thrombocytic dystrophy). AB - Bernard-Soulier syndrome (BSS), also known as Hemorrhagiparous thrombocytic dystrophy, is a hereditary bleeding disorder affecting the megakaryocyte/platelet lineage and characterized by bleeding tendency, giant blood platelets and low platelet counts. This syndrome is extremely rare as only approximately 100 cases have been reported in the literature. Clinical manifestations usually include purpura, epistaxis, menorrhagia, gingival and gastrointestinal bleeding. The syndrome is transmitted as an autosomal recessive trait. The underlying defect is a deficiency or dysfunction of the glycoprotein GPIb-V-IX complex, a platelet restricted multisubunit receptor required for normal primary hemostasis. The GPIb V-IX complex binds von Willebrand factor, allowing platelet adhesion and platelet plug formation at sites of vascular injury. Genes coding for the four subunits of the receptor, GPIBA, GPIBB, GP5 and GP9, map to chromosomes 17p12, 22q11.2, 3q29, and 3q21, respectively. Defects have been identified in GPIBA, GPIBB, and GP9 but not in GP5. Diagnosis is based on a prolonged skin bleeding time, the presence of a small number of very large platelets (macrothrombocytopenia), defective ristocetin-induced platelet agglutination and low or absent expression of the GPIb-V-IX complex. Prothrombin consumption is markedly reduced. The prognosis is usually good with adequate supportive care but severe bleeding episodes can occur with menses, trauma and surgical procedures. Treatment of bleeding or prophylaxis during surgical procedures usually requires platelet transfusion. PMID- 17109745 TI - Exogenous glucosamine globally protects chondrocytes from the arthritogenic effects of IL-1beta. AB - The effects of exogenous glucosamine on the biology of articular chondrocytes were determined by examining global transcription patterns under normal culture conditions and following challenge with IL-1beta. Chondrocytes isolated from the cartilage of rats were cultured in several flasks either alone or in the presence of 20 mM glucosamine. Six hours later, one-half of the cultures of each group were challenged with 10 ng/ml IL-1beta. Fourteen hours after this challenge, RNA was extracted from each culture individually and used to probe microarray chips corresponding to the entire rat genome. Glucosamine alone had no observable stimulatory effect on the transcription of primary cartilage matrix genes, such as aggrecan, collagen type II, or genes involved in glycosaminoglycan synthesis; however, glucosamine proved to be a potent, broad-spectrum inhibitor of IL-1beta. Of the 2,813 genes whose transcription was altered by IL-1beta stimulation (P < 0.0001), glucosamine significantly blocked the response in 2,055 (approximately 73%). Glucosamine fully protected the chondrocytes from IL-1-induced expression of inflammatory cytokines, chemokines, and growth factors as well as proteins involved in prostaglandin E2 and nitric oxide synthesis. It also blocked the IL-1 induced expression of matrix-specific proteases such as MMP-3, MMP-9, MMP-10, MMP 12, and ADAMTS-1. The concentrations of IL-1 and glucosamine used in these assays were supraphysiological and were not representative of the arthritic joint following oral consumption of glucosamine. They suggest, however, that the potential benefit of glucosamine in osteoarthritis is not related to cartilage matrix biosynthesis, but is more probably related to its ability to globally inhibit the deleterious effects of IL-1beta signaling. These results suggest that glucosamine, if administered effectively, may indeed have anti-arthritic properties, but primarily as an anti-inflammatory agent. PMID- 17109746 TI - Reliability and validity of the ESRD Symptom Checklist--Transplantation Module in Norwegian kidney transplant recipients. AB - BACKGROUND: The aim of the study was to validate the Norwegian version of a self administered 43-item questionnaire designed to assess quality of life in kidney transplant recipients, the End-Stage Renal Disease Symptom Checklist- Transplantation Module (ESRD-SCL). METHODS: In total, 53 kidney transplant recipients from one university-affiliated hospital responded to a questionnaire including the ESRD-SCL and the Short Form 36 (SF-36). We assessed internal consistency reliability and test-retest reliability with 2 weeks between assessments. Construct validity was assessed by correlations of the ESRD-SCL subscales with related and unrelated SF-36 scales, demographic, and clinical characteristics. RESULTS: Subscales of the ESRD-SCL showed good internal consistency reliability (Cronbach's = 0.72-0.81) and for the aggregate total scale alpha was 0.94. Test-retest reliability median 14 days apart was excellent with intraclass coefficients ranging from 0.87 to 0.95. The pattern of correlations of the ESRD-SCL scales with related and unrelated scales SF-36 scales and demographic and clinical characteristics gave support to the construct validity of the ESRD-SCL. CONCLUSION: The Norwegian translation of the ESRD-SCL showed satisfactory internal consistency reliability, test-retest reliability and construct validity, at the level of the original German version. PMID- 17109747 TI - Modelling multiple hospital outcomes: the impact of small area and primary care practice variation. AB - BACKGROUND: Appropriate management of care--for example, avoiding unnecessary attendances at, or admissions to, hospital emergency units when they could be handled in primary care--is an important part of health strategy. However, some variations in these outcomes could be due to genuine variations in health need. This paper proposes a new method of explaining variations in hospital utilisation across small areas and the general practices (GPs) responsible for patient primary care. By controlling for the influence of true need on such variations, one may identify remaining sources of excess emergency attendances and admissions, both at area and practice level, that may be related to the quality, resourcing or organisation of care. The present paper accordingly develops a methodology that recognises the interplay between population mix factors (health need) and primary care factors (e.g. referral thresholds), that allows for unobserved influences on hospitalisation usage, and that also reflects interdependence between hospital outcomes. A case study considers relativities in attendance and admission rates at a North London hospital involving 149 small areas and 53 GP practices. RESULTS: A fixed effects model shows variations in attendances and admissions are significantly related (positively) to area and practice need, and nursing home patients, and related (negatively) to primary care access and distance of patient homes from the hospital. Modelling the impact of known factors alone is not sufficient to produce a satisfactory fit to the observations, and random effects at area and practice level are needed to improve fit and account for overdispersion. CONCLUSION: The case study finds variation in attendance and admission rates across areas and practices after controlling for need, and remaining differences between practices may be attributable to referral behaviour unrelated to need, or to staffing, resourcing, and access issues. In managerial terms, the analysis points to the utility of formal statistical analysis of hospitalisation rates as a prelude to non-statistical investigation of primary care resourcing and organisation. For example, there may be implications for the location of staff involved in community management of chronic conditions; health managers may also investigate whether some practices have unusual populations (homeless, asylum seekers, students) that explain different hospital use patterns. PMID- 17109748 TI - Surgery is more cost-effective than splinting for carpal tunnel syndrome in the Netherlands: results of an economic evaluation alongside a randomized controlled trial. AB - BACKGROUND: Carpal tunnel syndrome (CTS) is a common disorder, often treated with surgery or wrist splinting. The objective of this economic evaluation alongside a randomized trial was to evaluate the cost-effectiveness of splinting and surgery for patients with CTS. METHODS: Patients at 13 neurological outpatient clinics with clinically and electrophysiologically confirmed idiopathic CTS were randomly allocated to splinting (n = 89) or surgery (n = 87). Clinical outcome measures included number of nights waking up due to symptoms, general improvement, severity of the main complaint, paraesthesia at night and during the day, and utility. The economic evaluation was performed from a societal perspective and involved all relevant costs. RESULTS: There were no differences in costs. The mean total costs per patient were in the surgery group EURO 2,126 compared to EURO 2,111 in the splint group. After 12 months, the success rate in the surgery group (92%) was significantly higher than in the splint group (72%). The acceptability curve showed that at a relatively low ceiling ratio of EURO 2,500 per patient there is a 90% probability that surgery is cost-effective. CONCLUSION: In the Netherlands, surgery is more cost-effective compared with splinting, and recommended as the preferred method of treatment for patients with CTS. PMID- 17109749 TI - ProCAT: a data analysis approach for protein microarrays. AB - Protein microarrays provide a versatile method for the analysis of many protein biochemical activities. Existing DNA microarray analytical methods do not translate to protein microarrays due to differences between the technologies. Here we report a new approach, ProCAT, which corrects for background bias and spatial artifacts, identifies significant signals, filters nonspecific spots, and normalizes the resulting signal to protein abundance. ProCAT provides a powerful and flexible new approach for analyzing many types of protein microarrays. PMID- 17109750 TI - High blood pressure in school children: prevalence and risk factors. AB - BACKGROUND: The purpose of this study was to determine the prevalence of high blood pressure (HBP) and associated risk factors in school children 8 to 13 years of age. METHODS: Elementary school children (n = 1,066) were examined. Associations between HBP, body mass index (BMI), gender, ethnicity, and acanthosis nigricans (AN) were investigated using a school based cross-sectional study. Blood pressure was measured and the 95th percentile was used to determine HBP. Comparisons between children with and without HBP were utilized. The crude and multiple logistic regression adjusted odds ratios were used as measures of association. RESULTS: Females, Hispanics, overweight children, and children with AN had an increased likelihood of HBP. Overweight children (BMI > or = 85th percentile) and those with AN were at least twice as likely to present with HBP after controlling for confounding factors. CONCLUSION: Twenty one percent of school children had HBP, especially the prevalence was higher among the overweight and Hispanic group. The association identified here can be used as independent markers for increased likelihood of HBP in children. PMID- 17109751 TI - Coronary by-pass for bad ventricle; adoption of "hybrid-pump" bypass. AB - BACKGROUND: The outcomes of on-pump and hybrid-pump bypass surgery in patients with depressed left ventricular function (EF<30%) were analyzed. METHODS: 109 patients with preoperative left ventricular ejection fraction of <30% and bypassable circumflex coronary disease were randomized in a double blind fashion to undergo hybrid-pump (combination of off-pump and on-pump) procedure (54 patients), or on-pump coronary bypass (55 patients). In patients who underwent hybrid-pump procedure only circumflex system was bypassed on-pump to shorten CPB and myocardial ischemic time. Pre- peri and postoperative variables were analyzed. RESULTS: Mean LVEF 24.4 +/- 4.8%. The patients in hybrid-pump group received less graft than others, but difference was not significant. Duration of the surgery was not different statistically between hybrid-pump and on-pump groups. A longer intraoperative duration of ischemia and extra corporeal circulation was found in on-pump group. Significant improvement in the postoperative course such as shorter mechanical ventilation, less catecholamines and IABP usage, less ICU and hospital stay, less stroke, less need for hemodyalisis and most importantly less hospital mortality was observed in hybrid pump group. CONCLUSION: Shortening the CPB and myocardial ischemic time and avoiding related problems, adoption of hybrid-pump strategy, in patients with severely impaired LVEF and bypassable circumflex coronary disease results in better outcome than conventional on-pump bypass. PMID- 17109752 TI - Predicting residue contacts using pragmatic correlated mutations method: reducing the false positives. AB - BACKGROUND: Predicting residues' contacts using primary amino acid sequence alone is an important task that can guide 3D structure modeling and can verify the quality of the predicted 3D structures. The correlated mutations (CM) method serves as the most promising approach and it has been used to predict amino acids pairs that are distant in the primary sequence but form contacts in the native 3D structure of homologous proteins. RESULTS: Here we report a new implementation of the CM method with an added set of selection rules (filters). The parameters of the algorithm were optimized against fifteen high resolution crystal structures with optimization criterion that maximized the confidentiality of the predictions. The optimization resulted in a true positive ratio (TPR) of 0.08 for the CM without filters and a TPR of 0.14 for the CM with filters. The protocol was further benchmarked against 65 high resolution structures that were not included in the optimization test. The benchmarking resulted in a TPR of 0.07 for the CM without filters and to a TPR of 0.09 for the CM with filters. CONCLUSION: Thus, the inclusion of selection rules resulted to an overall improvement of 30%. In addition, the pair-wise comparison of TPR for each protein without and with filters resulted in an average improvement of 1.7. The methodology was implemented into a web server http://www.ces.clemson.edu/compbio/recon that is freely available to the public. The purpose of this implementation is to provide the 3D structure predictors with a tool that can help with ranking alternative models by satisfying the largest number of predicted contacts, as well as it can provide a confidence score for contacts in cases where structure is known. PMID- 17109753 TI - Outcome of postoperative critically ill patients with heparin-induced thrombocytopenia: an observational retrospective case-control study. AB - INTRODUCTION: Heparin-induced thrombocytopenia (HIT) is described as a decrease in platelet count associated with heparin administration and is an immune mediated adverse drug reaction that can cause both arterial and venous thromboses. It can be a life-threatening complication of heparin exposure. Little data concerning incidence, predisposing factors, or outcome in critically ill surgical patients are available. METHODS: All critically ill, postoperative patients admitted between January 1, 2000, and December 31, 2001, to a surgical intensive care unit (SICU) who tested positive by an enzyme-linked immunosorbent assay for the HIT antibody (HPIA; Diagnostica Stago, Inc., Parsippany, NJ, USA) were identified. Patient risk factors and outcomes were abstracted retrospectively from the medical record and compared with those from control patients matched for age, gender, diagnosis, severity of illness, and date of SICU admission. RESULTS: Two hundred and ten patients out of 2,046 patients (10%) admitted to the SICU had HIT assays performed. Nineteen patients (0.9% of admissions; 9% of tested individuals) had positive tests. HIT-antibody-positive patients, compared with 19 matched controls, had an increased risk of death or major thrombotic complications (37% versus 10%; P < 0.05) and prolonged length of intensive care unit (ICU) stay (20 days versus 10 days; P < 0.05). Exposure to heparin via intravascular flushes alone was sufficient to generate HIT antibodies in 12 of 19 (63%) patients. Five patients received platelet transfusions after the diagnosis of HIT was known; four of these patients died. CONCLUSION: Heparin flushes were the most common cause of HIT in this study. HIT-antibody-positive patients had an increased risk of death or major complications and a prolonged length of ICU stay. Platelet transfusions often were administered despite a positive HIT test result and were associated with a high mortality rate. Treatment algorithms that minimize exposure to heparin and contraindicate platelet transfusions merit further study. PMID- 17109754 TI - Wildtype epidermal growth factor receptor (Egfr) is not required for daily locomotor or masking behavior in mice. AB - BACKGROUND: Recent studies have implicated the epidermal growth factor receptor (EGFR) within the subparaventricular zone as being a major mediator of locomotor and masking behaviors in mice. The results were based on small cohorts of mice homozygous for the hypomorphic Egfrwa2 allele on a mixed, genetically uncontrolled background, and on intraventricular infusion of exogenous EGFR ligands. Subsequenlty, a larger study using the same genetically mixed background failed to replicate the original findings. Since both previous approaches were susceptible to experimental artifacts related to an uncontrolled genetic background, we analyzed the locomotor behaviors in Egfrwa2 mutant mice on genetically defined, congenic backgrounds. METHODS: Mice carrying the Egfrwa2 hypomorphic allele were bred to congenicity by backcrossing greater than ten generations onto C57BL/6J and 129S1/SvImJ genetic backgrounds. Homozygous Egfrwa2 mutant and wildtype littermates were evaluated for defects in locomotor and masking behaviors. RESULTS: Mice homozygous for Egfrwa2 showed normal daily locomotor activity and masking indistinguishable from wildtype littermates at two light intensities (200-300 lux and 400-500 lux). CONCLUSION: Our results demonstrate that reduced EGFR activity alone is insufficient to perturb locomotor and masking behaviors in mice. Our results also suggest that other uncontrolled genetic or environmental parameters confounded previous experiments linking EGFR activity to daily locomotor activity and provide a cautionary tale for genetically uncontrolled studies. PMID- 17109755 TI - A comparative analysis of the information content in long and short SAGE libraries. AB - BACKGROUND: Serial Analysis of Gene Expression (SAGE) is a powerful tool to determine gene expression profiles. Two types of SAGE libraries, ShortSAGE and LongSAGE, are classified based on the length of the SAGE tag (10 vs. 17 basepairs). LongSAGE libraries are thought to be more useful than ShortSAGE libraries, but their information content has not been widely compared. To dissect the differences between these two types of libraries, we utilized four libraries (two LongSAGE and two ShortSAGE libraries) generated from the hippocampus of Alzheimer and control samples. In addition, we generated two additional short SAGE libraries, the truncated long SAGE libraries (tSAGE), from LongSAGE libraries by deleting seven 5' basepairs from each LongSAGE tag. RESULTS: One problem that occurred in the SAGE study is that individual tags may have matched to multiple different genes - due to the short length of a tag. We found that the LongSAGE tag maps up to 15 UniGene clusters, while the ShortSAGE and tSAGE tags map up to 279 UniGene clusters. Both long and short SAGE libraries exhibit a large number of orphan tags (no gene information in UniGene), implying the limitation of the UniGene database. Among 100 orphan LongSAGE tags, the complete sequences (17 basepairs) of nine orphan tags match to 17 genomic sequences; four of the orphan tags match to a single genomic sequence. Our data show the potential to resolve 4-9% of orphan LongSAGE tags. Finally, among 400 tSAGE tags showing significant differential expression between AD and control, 79 tags (19.8%) were derived from multiple non-significant LongSAGE tags, implying the false positive results. CONCLUSION: Our data show that LongSAGE tags have high specificity in gene mapping compared to ShortSAGE tags. LongSAGE tags show an advantage over ShortSAGE in identifying novel genes by BLAST analysis. Most importantly, the chances of obtaining false positive results are higher for ShortSAGE than LongSAGE libraries due to their specificity in gene mapping. Therefore, it is recommended that the number of corresponding UniGene clusters (gene or ESTs) of a tag for prioritizing the significant results be considered. PMID- 17109756 TI - The centrosome and cell proliferation. AB - Centrosomes are frequently amplified in cancer cells. Increased numbers of centrosomes can give rise to multipolar spindles in mitosis, and thereby lead to the formation of aneuploid daughter cells. However, whether centrosome amplification is a cause or a consequence of cancer is unclear. In contrast, loss of a functional centrosome has been shown to lead to cell cycle arrest. In this review, the potential mechanisms underlying centrosome amplification and centrosome-dependent cell cycle regulation are discussed. PMID- 17109757 TI - EXMOTIF: efficient structured motif extraction. AB - BACKGROUND: Extracting motifs from sequences is a mainstay of bioinformatics. We look at the problem of mining structured motifs, which allow variable length gaps between simple motif components. We propose an efficient algorithm, called EXMOTIF, that given some sequence(s), and a structured motif template, extracts all frequent structured motifs that have quorum q. Potential applications of our method include the extraction of single/composite regulatory binding sites in DNA sequences. RESULTS: EXMOTIF is efficient in terms of both time and space and is shown empirically to outperform RISO, a state-of-the-art algorithm. It is also successful in finding potential single/composite transcription factor binding sites. CONCLUSION: EXMOTIF is a useful and efficient tool in discovering structured motifs, especially in DNA sequences. The algorithm is available as open-source at: http://www.cs.rpi.edu/~zaki/software/exMotif/. PMID- 17109758 TI - Hyaluronan- and RNA-binding deubiquitinating enzymes of USP17 family members associated with cell viability. AB - BACKGROUND: Protein degradation by the ubiquitin system plays a crucial role in numerous cellular signaling pathways. Deubiquitination, a reversal of ubiquitination, has been recognized as an important regulatory step in the ubiquitin-dependent degradation pathway. RESULTS: While identifying putative ubiquitin specific protease (USP) enzymes that contain a conserved Asp (I) domain in humans, 4 USP17 subfamily members, highly homologous to DUB-3, have been found (USP17K, USP17L, USP17M, and USP17N), from human chorionic villi. Expression analysis showed that USP17 transcripts are highly expressed in the heart, liver, and pancreas and are expressed moderately in various human cancerous cell lines. Amino acid sequence analysis revealed that they contain the highly conserved Cys, His, and Asp domains which are responsible for the deubiquitinating activity. Biochemical enzyme assays indicated that they have deubiquitinating activity. Interestingly, the sequence analysis showed that these proteins, with exception of USP17N, contain the putative hyaluronan/RNA binding motifs, and cetylpyridinium chloride (CPC)-precipitation analysis confirmed the association between these proteins and intracellular hyaluronan and RNA. CONCLUSION: Here, we report that the overexpression of these proteins, with exception of USP17N, leads to apoptosis, suggesting that the hyaluronan and RNA binding motifs in these enzymes play an important role in regulating signal transduction involved in cell death. PMID- 17109759 TI - In silico and microarray-based genomic approaches to identifying potential vaccine candidates against Leptospira interrogans. AB - BACKGROUND: Currently available vaccines against leptospirosis are of low efficacy, have an unacceptable side-effect profile, do not induce long-term protection, and provide no cross-protection against the different serovars of pathogenic leptospira. The current major focus in leptospirosis research is to discover conserved protective antigens that may elicit longer-term protection against a broad range of Leptospira. There is a need to screen vaccine candidate genes in the genome of Leptospira interrogans. RESULTS: Bioinformatics, comparative genomic hybridization (CGH) analysis and transcriptional analysis were used to identify vaccine candidates in the genome of L. interrogans serovar Lai strain #56601. Of a total of 4727 open reading frames (ORFs), 616 genes were predicted to encode surface-exposed proteins by P-CLASSIFIER combined with signal peptide prediction, alpha-helix transmembrane topology prediction, integral beta barrel outer membrane protein and lipoprotein prediction, as well as by retaining the genes shared by the two sequenced L. interrogans genomes and by subtracting genes with human homologues. A DNA microarray of L. interrogans strain #56601 was constructed for CGH analysis and transcriptome analysis in vitro. Three hundred and seven differential genes were identified in ten pathogenic serovars by CGH; 1427 genes had high transcriptional levels (Cy3 signal > or = 342 and Cy5 signal > or = 363.5, respectively). There were 565 genes in the intersection between the set encoding surface-exposed proteins and the set of 307 differential genes. The number of genes in the intersection between this set of 565 and the set of 1427 highly transcriptionally active genes was 226. These 226 genes were thus identified as putative vaccine candidates. The proteins encoded by these genes are not only potentially surface-exposed in the bacterium, but also conserved in two sequenced L. interrogans. Moreover, these genes are conserved among ten epidemic serovars in China and have high transcriptional levels in vitro. CONCLUSION: Of the 4727 ORFs in the genome of L. interrogans, 226 genes were identified as vaccine candidates by bioinformatics, CGH and transcriptional analysis on the basis of the theory of reverse vaccinology. The proteins encoded by these genes might be useful as vaccine candidates as well as for diagnosis of leptospirosis. PMID- 17109760 TI - Construction of a 7-fold BAC library and cytogenetic mapping of 10 genes in the giant panda (Ailuropoda melanoleuca). AB - BACKGROUND: The giant panda, one of the most primitive carnivores, is an endangered animal. Although it has been the subject of many interesting studies during recent years, little is known about its genome. In order to promote research on this genome, a bacterial artificial chromosome (BAC) library of the giant panda was constructed in this study. RESULTS: This BAC library contains 198,844 clones with an average insert size of 108 kb, which represents approximately seven equivalents of the giant panda haploid genome. Screening the library with 15 genes and 8 microsatellite markers demonstrates that it is representative and has good genome coverage. Furthermore, ten BAC clones harbouring AGXT, GHR, FSHR, IRBP, SOX14, TTR, BDNF, NT-4, LH and ZFX1 were mapped to 8 pairs of giant panda chromosomes by fluorescence in situ hybridization (FISH). CONCLUSION: This is the first large-insert genomic DNA library for the giant panda, and will contribute to understanding this endangered species in the areas of genome sequencing, physical mapping, gene cloning and comparative genomic studies. We also identified the physical locations of ten genes on their relative chromosomes by FISH, providing a preliminary framework for further development of a high resolution cytogenetic map of the giant panda. PMID- 17109761 TI - Effect of changes in contractility on the index of myocardial performance in the dysfunctional left ventricle. AB - BACKGROUND: The index of myocardial performance has prognostic power in patients with cardiomyopathy and following myocardial infarction. As the index of myocardial performance has been shown to be preload and afterload dependent, the effect of altering contractility on IMP and its components with left ventricular dysfunction has been incompletely delineated. METHODS: Chronic left ventricular dysfunction was induced in 10 canines using coronary microsphere embolization. Each dog was instrumented and imaged with 2D echo and Doppler. At the same atrially paced rate, contractility was increased with a dobutamine infusion and then following 4 weeks of oral digoxin. RESULTS: With chronic left ventricular dysfunction, a reduced left ventricular ejection fraction (42 +/- 3%, p < 0.001) and increased index of myocardial performance (0.58 +/- 0.17, p < 0.01) due to isovolumic contraction time lengthening and shortened left ventricular ejection time were noted. Dobutamine increased ejection fraction (p < 0.001), reduced left ventricular end diastolic pressure (p < 0.01), and reduced the index of myocardial performance (0.33 +/- 0.17, p < 0.001) due to isovolumic contraction time, isovolumic relaxation time, and left ventricular ejection time shortening. Digoxin increased ejection fraction (p < 0.05), reduced left ventricular end diastolic pressure (p < 0.05), and reduced the index of myocardial performance (0.42 +/- 0.13, p < 0.01) due to isovolumic contraction time shortening (p < 0.001). Both dobutamine and digoxin lengthened the diastolic filling period (p < 0.01). CONCLUSION: Increased inotropy with digoxin and dobutamine reduced the index of myocardial performance in dogs with left ventricular dysfunction. Shortened isovolumic contraction time, increased diastolic filling period, and reduced left ventricular end diastolic pressure with digoxin may provide insight into its efficacy in heart failure. PMID- 17109762 TI - Leptin concentration in breast milk and its relationship to duration of lactation and hormonal status. AB - BACKGROUND: Leptin, a hormone present in breast milk, is involved in energy regulation and metabolism. The objectives of this study were to assess leptin concentrations in breast milk during the first 180 days postpartum, and to determine the relationship between the concentrations of milk leptin and circulating hormone levels in lactating women. METHODS: Between April 2005 and January 2006, blood and breast milk samples were collected from 160 breastfeeding women enrolled either in the first three days (n = 37; colostrum), days 4-14 (n = 27; transitional milk), days 15-30 (n = 16; early mature milk), days 31-90 (n = 37; mature milk) or days 91-180 (n = 43; late mature milk) postpartum. Milk and serum leptin levels were measured by immunoradiometric assay. Cortisol was measured by radioimmunoassay method. Serum insulin, estradiol, prolactin and thyroxine were measured by chemiluminescent immunometric method. RESULTS: Leptin concentrations in breast milk were highest (3.28 +/- 0.41 ng/ml) in colostrum, decreased during the first 180 days of lactation, showing a significant inverse relation (r = -0.694, p < 0.001) with the days of lactation. Colostrum leptin concentrations correlated with maternal serum leptin (r = 0.425, p < 0.01), cortisol (r = 0.549, p < 0.01) and thyroxine (r = -0.530, p < 0.01). Mature milk leptin concentrations correlated with maternal serum leptin (r = 0.547, p < 0.001), insulin (r = 0.331, p < 0.05) and thyroxine (r = -0.329, p < 0.01). Serum leptin concentrations correlated with serum insulin (r = 0.648, p < 0.001), estradiol (r = 0.639, p < 0.001), prolactin (r = 0.530, p < 0.001) and thyroxine (r = -0.327, p < 0.05) concentrations during days 1-3 postpartum. During 15-180 postpartum days, serum leptin concentrations correlated with serum insulin (r = 0.271, p < 0.01), and thyroxine (r = -0.345, p < 0.001). CONCLUSION: Leptin concentrations in breast milk decrease with time during lactation and show significant relationships with other maternal hormones. PMID- 17109763 TI - Vulnerability to HIV infection among sex worker and non-sex worker female injecting drug users in Dhaka, Bangladesh: evidence from the baseline survey of a cohort study. AB - BACKGROUND: Very little is known about female injecting drug users (IDU) in Bangladesh but anecdotal evidence suggests that they are hidden and very vulnerable to HIV through both their injection sharing and sexual risk behaviors. In order to better understand the risks and vulnerability to HIV of female IDU, a cohort study was initiated through which HIV prevalence and risk behaviors was determined. METHODS: All female IDU (those who had injected in the last six months and were 15 years or older) who could be identified from three cities in the Dhaka region were enrolled at the baseline of a cohort study. The study was designed to determine risk behaviors through interviews using a semi-structured questionnaire and measure prevalence of HIV, hepatitis C and syphilis semiannually. At the baseline of the cohort study 130 female IDU were recruited and female IDU selling sex in the last year (sex workers) versus those not selling sex (non-sex workers) were compared using descriptive statistics and logistic regression. RESULTS: Of the 130 female IDU enrolled 82 were sex workers and 48 were non-sex workers. None had HIV but more sex workers (60%) had lifetime syphilis than non-sex workers (37%). Fewer sex worker than non-sex worker IDU lived with families (54.9% and 81.3% respectively), but more reported lending needles/syringes (29.3% and 14.6% respectively) and sharing other injection paraphernalia (74.4% and 56.3% respectively) in the past six months. Although more sex workers used condoms during last sex than non-sex workers (74.4% and 43.3% respectively), more reported anal sex (15.9% and 2.1% respectively) and serial sex with multiple partners (70.7% and 0% respectively). Lifetime sexual violence and being jailed in the last year was more common in sex workers. CONCLUSION: Female IDU are vulnerable to HIV through their injection and sexual risk behaviors and sex worker IDU appear especially vulnerable. Services such as needle exchange programs should become more comprehensive to address the needs of female IDU. PMID- 17109764 TI - Using local gene expression similarities to discover regulatory binding site modules. AB - BACKGROUND: We present an approach designed to identify gene regulation patterns using sequence and expression data collected for Saccharomyces cerevisae. Our main goal is to relate the combinations of transcription factor binding sites (also referred to as binding site modules) identified in gene promoters to the expression of these genes. The novel aspects include local expression similarity clustering and an exact IF-THEN rule inference algorithm. We also provide a method of rule generalization to include genes with unknown expression profiles. RESULTS: We have implemented the proposed framework and tested it on publicly available datasets from yeast S. cerevisae. The testing procedure consists of thorough statistical analyses of the groups of genes matching the rules we infer from expression data against known sets of co-regulated genes. For this purpose we have used published ChIP-Chip data and Gene Ontology annotations. In order to make these tests more objective we compare our results with recently published similar studies. CONCLUSION: Results we obtain show that local expression similarity clustering greatly enhances overall quality of the derived rules, both in terms of enrichment of Gene Ontology functional annotation and coherence with ChIP-Chip binding data. Our approach thus provides reliable hypotheses on co regulation that can be experimentally verified. An important feature of the method is its reliance only on widely accessible sequence and expression data. The same procedure can be easily applied to other microbial organisms. PMID- 17109765 TI - Visual setup of logical models of signaling and regulatory networks with ProMoT. AB - BACKGROUND: The analysis of biochemical networks using a logical (Boolean) description is an important approach in Systems Biology. Recently, new methods have been proposed to analyze large signaling and regulatory networks using this formalism. Even though there is a large number of tools to set up models describing biological networks using a biochemical (kinetic) formalism, however, they do not support logical models. RESULTS: Herein we present a flexible framework for setting up large logical models in a visual manner with the software tool ProMoT. An easily extendible library, ProMoT's inherent modularity and object-oriented concept as well as adaptive visualization techniques provide a versatile environment. Both the graphical and the textual description of the logical model can be exported to different formats. CONCLUSION: New features of ProMoT facilitate an efficient set-up of large Boolean models of biochemical interaction networks. The modeling environment is flexible; it can easily be adapted to specific requirements, and new extensions can be introduced. ProMoT is freely available from http://www.mpi-magdeburg.mpg.de/projects/promot/. PMID- 17109766 TI - Transcription factor 7-like 2 (TCF7L2) variant is associated with familial breast cancer risk: a case-control study. AB - BACKGROUND: The transcription factor 7-like 2 (TCF7L2) is a critical component of the Wnt/beta-catenin pathway. Aberrant TCF7L2 expression modifies Wnt signaling and mediates oncogenic effects through the upregulation of c-MYC and cyclin D. Genetic alterations in TCF7L2 may therefore affect cancer risk. Recently, TCF7L2 variants, including the microsatellite marker DG10S478 and the nearly perfectly linked SNP rs12233372, were identified to associate with type 2 diabetes. METHODS: We investigated the effect of the TCF7L2 rs12255372 variant on familial breast cancer (BC) risk by means of TaqMan allelic discrimination, analyzing BRCA1/2 mutation-negative index patients of 592 German BC families and 735 control individuals. RESULTS: The T allele of rs12255372 showed an association with borderline significance (OR = 1.19, 95% C.I. = 1.01-1.42, P = 0.04), and the Cochran-Armitage test for trend revealed an allele dose-dependent association of rs12255372 with BC risk (Ptrend = 0.04). CONCLUSION: Our results suggest a possible influence of TCF7L2 rs12255372 on the risk of familial BC. PMID- 17109767 TI - A simulation model to assess herd-level intervention strategies against E. coli O157. AB - A simulation model of a herd of grazing cattle, which has been developed to provide insight into the infection dynamics of E. coli O157 is described. The spatially explicit model enables the modelling of the infection transmission processes to be realistically addressed under field management conditions. The model is used to explore the efficacy of various potential control strategies in reducing the levels of within-herd infection. These measures include restricting the size of herds, niche engineering, improving housing hygiene and vaccination. While a vaccination strategy remains a hypothetical option, it has the potential to be particularly effective. It is likely that the most successful strategy will involve the implementation of a combination of measures. PMID- 17109768 TI - Review and critical discussion of assumptions and modelling options to study the spread of the bovine viral diarrhoea virus (BVDV) within a cattle herd. AB - Relevance of epidemiological models depends on assumptions on the population structure and dynamics, on the biology of the host-parasite interaction, and on mathematical modelling. In this paper we reviewed published models of the bovine viral diarrhoea virus (BVDV) spread within a herd. Modelling options and assumptions on herd dynamics and BVDV transmission were discussed. A cattle herd is a population with a controlled size. Animals are separated into subgroups according to their age or their physiological status inducing heterogeneity of horizontal transmission. Complexity of models results from: (1) horizontal and vertical virus transmission, (2) birth of persistently infected animals, (3) excretion by transiently and persistently infected animals. Areas where there was a lack of knowledge were identified. Assumptions on the force of infection used to model the horizontal virus transmission were presented and discussed. We proposed possible ways of improving models (e.g. force of infection, validation) and essential model features for further BVDV models. PMID- 17109769 TI - Specific detection of Campylobacter jejuni from faeces using single nucleotide polymorphisms. AB - Specimens of human faeces were tested by a rapid strategy for detection of Campylobacter jejuni lineages by the presence of specific single nucleotide polymorphisms (SNPs) based on the C. jejuni multi locus sequence typing (MLST) scheme. This strategy was derived from analysis of the MLST databases to identify clonal complex specific SNPs followed by the design of real-time PCR assays to enable identification of six major C. jejuni clonal complexes associated with cases of human infection. The objective was to use the MLST SNP-based assays for the direct detection of C. jejuni by clonal complex from specimens of human faeces, and then confirm the accuracy of the clonal complex designation from the SNP-based assays by performing MLST on the cultured faecal material, this targeted at determining the validity of direct molecular specimen identification. Results showed it was possible to identify 38% of the isolates to one of the six major MLST clonal complexes using a rapid DNA extraction method directly from faeces in under 3 h. This method provides a novel strategy for the use of real time PCR for detection and characterization beyond species level, supplying real time epidemiological data, which is comparable with MLST results. PMID- 17109770 TI - Gastroenteritis associated with accidental contamination of drinking water with partially treated water. AB - Due to human error, drinking water supplied to a new housing estate in The Netherlands was contaminated with grey water. The cohort of 921 accidentally exposed households (area A) had a higher attack rate for diarrhoea (54.1%) than a non-exposed cohort of 1529 households from an adjacent area (B) (24%) (RR 2.3, 95% CI 1.9-2.7). Household water score showed a dose-response with illness, in both areas A and B. For each 1000 inhabitants, 19.8 cases in area A, 7.0 cases in control area B (RRAB 2.2, 95% CI 1.3-3.8) and 3.3 cases in a more distant control area C (RRAC 4.6, 95% CI 2.7-8.0) were diagnosed with gastroenteritis by their general practitioner. A gastroenteritis outbreak associated with consumption of contaminated drinking water was observed in the exposed area. The use of grey water was banned in 2003, with the exception of rainwater use for flushing toilets. The risk of rainwater use is currently being investigated. PMID- 17109771 TI - More than one variant of Listeria monocytogenes isolated from each of two human cases of invasive listeriosis. AB - Two variants of Listeria monocytogenes were isolated from blood cultures from each of two patients with listeriosis. Each variant displayed a two-band difference in DNA profile from the other by pulsed-field gel electrophoresis. Although this difference in profile is insufficient to distinguish clearly between the variants, the possibility of co-infection with different strains of L. monocytogenes needs to be considered. We suggest that more than one colony should be selected for molecular typing to aid interpretation during investigation of the sources and routes of Listeria infection. PMID- 17109772 TI - Rotavirus VP7, VP4 and VP6 genotypes co-circulating in Tehran, Iran, between 2003 and 2004. AB - Rotaviruses were detected by enzyme-linked immunosorbent assay (ELISA) in 92 out of 374 faecal samples collected between November 2003 and October 2004 at the Markaz Tebbi Koudakan Hospital, Tehran, Iran, from children aged 6 months to 5 years. Analysis of clinical and disease severity data showed a significant association between rotavirus infection and diarrhoea, vomiting and severe dehydration. Ninety-two samples (64 rotavirus ELISA-positive and 28 ELISA negative samples) were sent to the Enteric Virus Unit, Virus Reference Department, Centre for Infection, Health Protection Agency, UK for rotavirus characterization by G-typing, P-typing and subgrouping (SG) using reverse transcriptase (RT)-PCR, semi-nested PCR and sequencing methods. In this study, both common and uncommon rotavirus genotypes were detected. The most prevalent types were G1P[8], SGII (59.2%) followed by G9P[8] SGII (15.5%) which has not been previously reported from Iran. Unusual genotypes G1P[10] SGI (2.8%) and G12P[8] SGII (1.4%) and strains derived from reassortment between common co circulating genotypes such as G1P[4] SGII represented 5.6% of strains. Mixed infections with combinations of G1+G4P[8] SGII and G1+G9P[8] SGII were also found. This contrasts with previous reports from Iran in which a small number of common rotavirus strains (G1 and G4) were found. This study highlights the need for continued surveillance and characterization of rotaviruses to take account of the rapid evolution and introduction of novel rotaviruses into the human population. PMID- 17109773 TI - Legionnaires' disease outbreak associated with a cruise liner, August 2003: epidemiological and microbiological findings. AB - Eight cases of Legionnaires' disease were identified among the 215 German passengers after a cruise to the Nordic Sea in August 2003. An unmatched case control study was conducted to identify risk factors and the source of infection. In total, eight passengers fulfilled the case definition, one of these died. Forty-two passengers served as controls. The attack rate was 4%. The mean age was 60 years for cases and 62 years for controls. Prolonged exposure to the spa pool seemed to be a risk factor of infection (OR 4.85, P=0.09). Legionella pneumophila serogroup 1, monoclonal antibody (mAb) subgroup 'Knoxville' was isolated from clinical and environmental samples. DNA sequence-based typing revealed that these isolates were indistinguishable from each other. The investigation showed the importance of an interdisciplinary approach of microbiology and epidemiology as not all sites on the ship that tested positive for L. pneumophila actually posed a relevant risk for the passengers. PMID- 17109774 TI - Low genetic diversity of the intrinsic OXA-51-like class D carbapenemases among Acinetobacter baumannii clinical isolates in Greece. AB - This study examined the geographical distribution and diversity of the intrinsic OXA-51-like class D carbapenemases among Acinetobacter baumannii clones recovered in three major Greek regions from 2000 to 2005. The blaOXA-66 allele was exclusively detected among clonally distinct A. baumannii isolates recovered in the regions of Thessaloniki and Larissa. This sequence was also the most widespread among A. baumannii isolates in Athens, while less frequent were blaOXA 69 and blaOXA-65 alleles. These findings highlight the high prevalence of a specific blaOXA-51-like allele in Greece, possibly indicating that our A. baumannii clones might have originated from a common ancestor. However, the possibility that blaOXA-51-like variants, with blaOXA-66 predominating, are widely disseminated among several unrelated A. baumannii strains cannot be excluded. PMID- 17109775 TI - Total antioxidant capacity of plasma in asymptomatic carrier state of Neisseria meningitidis. AB - Reduction of the antioxidant capacity of plasma has been linked with the impairment of an effective immune response and so we hypothesized that the carriage rate of Neisseria meningitidis in asymptomatic subjects might correlate with the levels of antioxidants in plasma. To this end we took pharyngeal swabs from 339 children in Marquesado Basic Health Zone, Granada, Spain and in addition determined the total antioxidant capacity (TAC) in plasma samples from these subjects. The overall prevalence of N. meningitidis carriage was 5.9% (mean age 7.1 years) with rates of 10.3% in children aged 3 < or =years, 3.9% between 4 and 7 years and 2.4% in older subjects. Plasma TAC for the < or =3-year-olds was 0.13 for carriers and 1.10 for non-carrier controls (P=0.04), 0.13 for carriers aged 4 7 years (controls 0.63) and 0.28 for carriers aged >7 years (controls 0.52). We analysed the association between TAC in plasma (<0.37 - 2 S.D.) and the carrier state of N. meningitidis. In the carrier state, the odds ratio for this association (TAC in plasma <0.25) was 8.44 (95% CI 1.5-48.9). These findings may suggest a reduced immune response in the host favourable to nasopharyngeal persistence of meningococci. PMID- 17109776 TI - Anxiety disorders and suicidal behaviours in adolescence and young adulthood: findings from a longitudinal study. AB - BACKGROUND: The aim of this study was to estimate the extent to which anxiety disorders contribute to an increase in suicidal behaviour after controlling for both observed and non-observed sources of confounding. METHOD: Data were collected from the Christchurch Health and Development Study (CHDS), a 25-year longitudinal study of over 1000 participants. Measures of anxiety disorders [phobia, generalized anxiety disorder (GAD), panic disorder], major depression (MD), substance use disorders, conduct/antisocial personality disorder, stressful life events, unemployment, and suicidal ideation/attempts for subjects aged 16 18, 18-21 and 21-25 years were used to fit random and fixed effects regression models of the associations between anxiety disorders and suicidal behaviours. RESULTS: Anxiety disorders were strongly associated with suicidal ideation/attempts. Any single anxiety disorder increased the odds of suicidal ideation by 7.96 times [95% confidence interval (CI) 5.69-11.13] and increased the rate of suicide attempts by 5.85 times (95% CI 3.66-9.32). Control for co occurring mental disorders, non-observed fixed confounding factors and life stress reduced these associations [suicidal ideation odds ratio (OR) 2.80, 95% CI 1.71-4.58; suicide attempts incidence rate ratio (IRR) 1.90, 95% CI 1.07-3.39]. Rates of suicidal behaviour also increased with the number of anxiety disorders. Estimates of the population attributable risk suggested that anxiety disorders accounted for 7-10% of the suicidality in the cohort. CONCLUSIONS: Anxiety disorders may be a risk factor for suicidality, even after controlling for confounding, with risks increasing with multiple anxiety disorders. Management of anxiety disorders may be an important component in strategies to reduce population rates of suicide. PMID- 17109777 TI - Emotional bias and waking salivary cortisol in relatives of patients with major depression. AB - BACKGROUND: Biases in the processing of emotional information have been shown to be abnormal in subjects with major depression, both during an episode and after full recovery. However, it is unclear whether these biases are a cause or an effect of the depression. This study set out to explore whether such biases represent a vulnerability factor for depression by looking at unaffected first degree relatives of those with major depressive disorder. We also measured waking salivary cortisol, as the regulation of the hypothalamo-pituitary-adrenal (HPA) axis is thought to be impaired in depressive disorder. METHOD: Twenty-five female relatives and 21 age-matched controls completed a facial expression recognition task, an emotional categorization task with positive and negative personality characteristics, and had their waking salivary cortisol measured on a work day and a non-work day. RESULTS: The depressed relative group was significantly faster to recognize facial expressions of fear than controls. The depressed relative group also showed significantly increased reaction time to recognize positive versus negative personality characteristics in the categorization task. There was no difference in waking salivary cortisol between groups, although there was an effect of work day versus non-work day. CONCLUSIONS: Subtle biases in the processing of emotional information may exist in the unaffected first degree relatives of those with depression. As such, this may represent a familial vulnerability factor to developing a depressive illness. PMID- 17109778 TI - Syndrome stability and psychological predictors of symptom severity in idiopathic environmental intolerance and somatoform disorders. AB - BACKGROUND: Previous studies suggest that idiopathic environmental intolerance (IEI) is a variant of somatoform disorders (SFDs) or the so-called functional somatic syndromes. Little is known, however, about the stability and the psychological predictors of IEI. METHOD: This prospective study examined the 1 year stability of somatic symptoms and IEI features in three diagnostic groups: 49 subjects with IEI, 43 subjects with SFD but without IEI, and 54 subjects (control group, CG) with neither IEI nor SFD. The predictive value of typical psychological predictors for somatization was tested using zero-order correlations and multiple linear regression analyses. RESULTS: Somatic symptoms and IEI features proved to be temporally stable over the 1-year follow-up period. The SFD and IEI groups scored significantly higher than CG on all measures of somatic symptoms and on questionnaires assessing psychological predictors for somatization. Measures of trait negative affectivity (NA), somatic symptom attribution and somatosensory amplification predicted somatic symptom severity within the IEI and SFD groups, both at baseline and 1 year later. The strongest predictors of IEI complaints in the IEI group were somatic attributions, followed by prominent cognitions of environmental threat and a tendency to focus on unpleasant bodily sensations and to consider them as pathological. CONCLUSIONS: IEI and SFD are highly stable conditions. In both SFD and IEI, NA and the processes of symptom perception, interpretation and attribution contribute substantially to the persistence of typically somatoform symptoms and IEI complaints. Treatment of IEI and SFD should address these psychological factors and mechanisms. PMID- 17109779 TI - Saposin-like proteins from the intestine of the blood-feeding hookworm, Ancylostoma caninum. AB - Hookworms feed on blood, utilizing haemoglobin for nutrition, growth and reproduction. The haemoglobin digestion cascade has been partially elucidated, but the process immediately preceding this event, haemolysis, has received considerably less attention. We have cloned and expressed Ancylostoma caninum mRNAs encoding 2 proteins belonging to the saposin-like protein (SAPLIP) family, termed Ac-slp-1 and Ac-slp-2. The open reading frames of SLP-1 and SLP-2 were used to identify expressed sequence tags encoding SAPLIPs from the 4 major clades of animal parasitic nematodes. Both Ac-slp-1 and slp-2 mRNAs were shown to be expressed in all life stages assessed, with slp-1 predominantly being expressed in third-stage larvae (L3) before and after activation with dog serum. Recombinant SLP-1 and SLP-2 were expressed in insect cells and used to raise specific antisera in mice. These antisera were used as probes in fluorescence microscopy to localize the anatomic expression sites of both proteins to small, punctate organelles or vesicles within the intestinal cells of adult worms; weak staining was detected on the microvillar brush border of the intestine. Using transmission electron microscopy, both proteins were localized to similar vesicles in the intestinal cells of the L3. Recombinant proteins contained C terminal purification tags that potentially precluded dimerization and possibly interfered with the subsequent detection of haemolytic activity. Their expression in the gut of the L3 and adult stages suggests a role for these hookworm SAPLIPs in the lysis of host cells during tissue migration and/or feeding. PMID- 17109780 TI - Biology of the schistosome lung-stage schistosomulum. AB - Past and more recent research has examined the ultrastructure, metabolism, cell biology, genomics and post-genomics of schistosome schistosomula. These areas are considered and discussed in this review with particular emphasis on (1) the early migration phases through the host, (2) interaction of the host immune response with the parasite surface, (3) glucose uptake mechanisms, and (4) defining the transcriptional profiles of lung-stage schistosomula compared with other developmental stages using microarrays. The microarray profiling studies suggest caution is required when considering the use of schistosomes obtained by in vitro means for molecular or biochemical studies. PMID- 17109781 TI - Reliability of the manual ability classification system for children with cerebral palsy. AB - The aim of this study was to determine the reliability of family and professional assessment of manual ability using the Manual Ability Classification System (MACS) for children with cerebral palsy (CP) in the UK. Families who were taking part in a study measuring the activities and participation of children with CP were invited to classify their child's manual ability using the MACS. Postal surveys were conducted with the families and health professionals nominated by the families. Perfect agreement was assessed as a percentage; chance-corrected agreement was measured using Cohen's kappa (kappa), and reliability was determined using the intraclass correlation coefficient (ICC). Families of 91/128 (71%) children responded to the survey (53 males, 38 females; mean age 9y 11mo [SD 1y 11mo], range 6-12y) out of whom 88 indicated a single MACS level. Seventy two children (82%) were classified with spastic CP, 12 (14%) with dyskinesia, two (2%) with ataxia, and two (2%) were not classified. There were 21, 27, 11, 10, and 19 children who were classified by their families in Gross Motor Function Classification System Levels I to V respectively; 14, 30, 18, 13, and 13 children classified by their families in MACS levels I to V. The survey of health professionals generated 60/71 (85%) responses from physiotherapists, 55/58 (93%) responses from paediatricians, and 21/24 (88%) responses from occupational therapists. There was perfect agreement between families and professionals for more than 50% of children; the indices of chance-corrected agreement ranged from kappa=0.3 to 0.5, and the reliability coefficients ranged from ICC 0.7 to 0.9. Indices of agreement and reliability between families and professionals were equivalent to those between different professionals. The MACS, therefore, offers a valid and reliable method for communicating about the manual ability of children with CP. Families and professionals may not always agree precisely on a MACS level, particularly if children's performance of manual tasks varies in different environments. PMID- 17109782 TI - Do the abilities of children with cerebral palsy explain their activities and participation? AB - The aim of this study was to use family-assessed instruments and details of children's impairments to explore factors affecting the activities and participation of children with cerebral palsy (CP). A postal survey was conducted with families of a geographically defined population of children with CP aged 6 to 12 years. Family-assessed indices of children's activities and participation were the Activities Scale for Kids (ASK) and Lifestyle Assessment Questionnaire (LAQ-CP). Families also assessed children's abilities using the Gross Motor Function and Manual Ability Classification Systems (GMFCS; MACS). Details of children's impairments were available from the 4Child epidemiological database and used with the GMFCS and MACS as explanatory variables in multiple regression analyses to identify their effect on children's activities and participation. Families of 175/314 (56%) children returned an assessment using the GMFCS and 129 (41%) children participated fully by returning all the questionnaires. Full participants (72 males, 57 females) did not differ from those who did not take part by their age, sex, CP characteristics, or associated impairments: GMFCS Level I-25, Level II-43, Level III-15, Level IV-14, Level V-23; MACS Level I-14, Level II-30, Level III-18, Level IV-13, Level V-13. Scores for the ASK and LAQ-CP Physical Independence and Mobility domains were predicted well by children's movement, manual, and intellectual disability, and also, to some extent, by the presence of seizures or speech problems. LAQ-CP domains for Economic and Clinical Burden and Social Integration were not well explained by children's abilities and impairments. Family assessment, therefore, offers a useful method for measuring children's activities and participation; however, currently available instruments do not fully represent all the domains in the International Classification of Functioning, Disability and Health. Children's abilities only partially explain their activities and participation. PMID- 17109783 TI - Differences in finger length ratio between males with autism, pervasive developmental disorder-not otherwise specified, ADHD, and anxiety disorders. AB - Children with autism have a relatively shorter index finger (2D) compared with their ring finger (4D). It is often presumed that the 2D:4D ratio is associated with fetal testosterone levels and that high fetal testosterone levels could play a role in the aetiology of autism. It is unknown whether this effect is specific to autism. In this study, 2D:4D ratios of 144 males aged 6 to 14 years (mean age 9y 1 mo [SD 1y 11 mo]) with psychiatric disorders were compared with those of 96 males aged 6 to 13 years from the general population (mean age 9y 1 mo [SD 1y 10 mo]). Psychiatric disorders were divided into autism/Asperger syndrome (n=24), pervasive developmental disorder-not otherwise specified (PDD-NOS; n=26), attention-deficit-hyperactivity disorder (ADHD)/oppositional defiant disorder (ODD; n=68), and anxiety disorders (n=26). Males with autism/Asperger syndrome (p<0.05) and ADHD/ODD (p<0.05) had significantly lower (though not significantly; p=0.52) ratios than males with an anxiety disorder, and males with autism/Asperger syndrome had lower ratios than those in the comparison group. These results indicated that higher fetal testosterone levels may play a role, not only in the origin of autism, but also in the aetiology of PDD-NOS and of ADHD/ODD. Males with anxiety disorders might have been exposed to lower prenatal testosterone levels. PMID- 17109784 TI - Specific postural support promotes variation in motor behaviour of infants with minor neurological dysfunction. AB - This study evaluated the effect of specific postural support on motor behaviour of infants with and without minor neurological dysfunction (MND). The following questions were addressed: (1) Does application of supportive pillows affect the time during which the infant exhibits general movements (GMs) or specific movements? We defined specific movements as movements of specific parts of the body that occur in a specific, recognizable way. (2) Does application of pillows improve the quality of GMs or the repertoire of specific movements? (3) Is a pillow effect affected by neurological condition? Forty healthy, term infants (16 males, 24 females; mean age 3.04 m [SD 1.24 mo], range 1-5 mo) participated in the study. Twenty were neurologically normal and 20 had MND. Spontaneous motor behaviour in a supine position was video-recorded for 180 seconds in four conditions applied in random order: support by a pillow in (1) the shoulder region, (2) the pelvic region, (3) the shoulder and pelvic region, or (4) no pillow support. Two independent assessors evaluated the quality of GMs. The other movement parameters were assessed with a computer program. Duration of movements was determined and a variation index, consisting of the number of different specific movements in a condition, was calculated. The presence of pillows did not affect the time spent in GMs, specific movements, or GM quality in either group. In neurologically normal infants the shoulder pillow with or without pelvic pillow induced an increase in the variation index (p<0.01), whereas in the infants with MND, all pillow conditions resulted in a substantial increase of the movement repertoire (p<0.001). Our results demonstrate that specific postural support promotes variation in motor behaviour of young infants. This is particularly true for infants with MND. PMID- 17109785 TI - Learning disabilities in children with neurofibromatosis type 1: subtypes, cognitive profile, and attention-deficit-hyperactivity disorder. AB - Cognitive deficits are the most common complication in children with neurofibromatosis type 1 (NF1), and academic achievement is broadly affected. There is a lack of consensus in the literature regarding the frequency of general and specific learning disabilities, which seems to be related to the lack of a consensus on diagnostic criteria. The present study examined the frequency of specific learning disabilities (SLDs) in NF1, using an intellect-achievement discrepancy for diagnosis, as well as general learning difficulty associated with a lowering of general intellectual ability. The cohort consisted of 81 children with NF1 (43 males, 38 females; mean age 11y 6mo [SD 2y 4mo]; range 8y-16y 9mo) and 49 comparison children (20 males, 29 females; mean age 12y [SD 2y 6mo]; range 8y 2mo-16y 8mo). Problems with academic achievement were present in 52% of children with NF1; however, only 20% of the children with NF1 were diagnosed with an SLD (32% had more general learning problems). Only males with NF1 were at significant risk for SLD, and Verbal IQ3-OH>2- or 4-OH. The coupling reactions with acylated glycosyl trichloroacetimidates as the donors usually give orthoesters as the intermediates specially when the coupling is carried out at slowed rates, and this is successfully used in regio- and stereoselective syntheses of oligosaccharides. Mannose and rhamnose orthoesters readily undergo O-2 (orthoester)C bond breaking, and this is used for synthesis of alpha-(1-->2) linked oligosaccharides. (1-->3)-Glucosylation is special since the rearrangement of its sugar orthoester intermediates can occur with either RO-(orthoester)C bond cleavage with formation of the dioxolenium ion leading to 1,2-trans linkage, or C 1-O-1 bond cleavage leading to 1,2-cis linkage, and this is dependent upon the structures of donor and acceptor that compose the orthoester. PMID- 17109836 TI - Application of saccharose as copper(II) ligand for electroless copper plating solutions. AB - Saccharose, forming sufficiently stable complexes with copper(II) ions in alkaline solutions, was found to be a suitable ligand for copper(II) chelating in alkaline (pH>12) electroless copper deposition solutions. Reduction of copper(II) saccharose complexes by hydrated formaldehyde was investigated and the copper deposits formed were characterized. The thickness of the compact copper coatings obtained under optimal operating conditions in 1h reaches ca. 2 microm at ambient temperature. The plating solutions were stable and no signs of Cu(II) reduction in the bulk solution were observed. Results were compared with those systems operating with other copper(II) ligands. PMID- 17109837 TI - The role of TGF-beta signaling in myocardial infarction and cardiac remodeling. AB - Transforming Growth Factor (TGF)-beta is markedly induced and rapidly activated in the infarcted myocardium. However, understanding of the exact role of TGF-beta signaling in the infarcted and remodeling heart has been hampered by the complex and unusual biology of TGF-beta activation and by the diversity of its effects eliciting multiple, and often opposing cellular responses. Experimental studies suggest that TGF-beta signaling may be crucial for repression of inflammatory gene synthesis in healing infarcts mediating resolution of the inflammatory infiltrate. In addition, TGF-beta may play an important role in modulating fibroblast phenotype and gene expression, promoting extracellular matrix deposition in the infarct by upregulating collagen and fibronectin synthesis and by decreasing matrix degradation through induction of protease inhibitors. TGF beta is also a key mediator in the pathogenesis of hypertrophic and dilative ventricular remodeling by stimulating cardiomyocyte growth and by inducing interstitial fibrosis. In this review we summarize the current knowledge on the role of TGF-beta in infarct healing and cardiac remodeling. PMID- 17109838 TI - Semaphorin 2a secreted by oenocytes signals through plexin B and plexin A to guide sensory axons in the Drosophila embryo. AB - The semaphorin gene family has been shown to play important roles in axonal guidance in both vertebrates and invertebrates. Both transmembrane (Sema1a, Sema1b, Sema5c) and secreted (Sema2a, Sema2b) forms of semaphorins exist in Drosophila. Two Sema receptors, plexins (Plex) A and B, have also been identified. Many questions remain concerning the axon guidance functions of the secreted semaphorins, including the identity of their receptors. We have used the well-characterized sensory system of the Drosophila embryo to address these problems. We find novel sensory axon defects in sema2a loss-of-function mutants in which particular axons misproject and follow inappropriate pathways to the CNS. plexB loss-of-function mutants show similar phenotypes to sema2a mutants and sema2a interacts genetically with plexB, supporting the hypothesis that Sema2a signals through PlexB receptors. Sema2a protein is expressed by larval oenocytes, a cluster of secretory cells in the lateral region of the embryo and the sema2a mutant phenotype can be rescued by driving Sema2a in these cells. Ablation of oenocytes results in sensory axon defects similar to the sema2a mutant phenotype. These data support a model in which Sema2a, while being secreted from oenocytes, acts in a highly localized fashion: It represses axon extension from the sensory neuron cell body, but only in regions in direct contact with oenocytes. PMID- 17109839 TI - A novel transforming growth factor-beta superfamily member expressed in gonadal somatic cells enhances primordial germ cell and spermatogonial proliferation in rainbow trout (Oncorhynchus mykiss). AB - Our understanding of the molecular mechanisms of primordial germ cell (PGC) proliferation in fish is rudimentary, but it is thought to be controlled by the surrounding somatic cells. We assumed that growth factors that are specifically involved in PGC proliferation are expressed predominantly in the surrounding genital ridge somatic cells. In order to isolate these growth factors, we compiled a complementary DNA (cDNA) subtractive library using cDNA from the genital ridges of 40-dpf rainbow trout embryos as the tester and cDNA from embryos without genital ridges as the driver. This approach identified a novel cytokine, designated gonadal soma-derived growth factor (GSDF), which is a member of the transforming growth factor (TGF)-beta superfamily. GSDF was expressed in the genital ridge somatic cells surrounding the PGCs during embryogenesis, and in both the granulosa and Sertoli cells at later stages. Inhibition of GSDF translation by antisense oligonucleotides suppressed PGC proliferation. Moreover, isolated testicular cells that were cultured with recombinant GSDF demonstrated dose-dependent proliferation of type-A spermatogonia; this effect was completely blocked by antiserum against GSDF. These results denote that GSDF, a novel member of the TGF-beta superfamily, plays an important role for proliferation of PGC and spermatogonia. PMID- 17109840 TI - A meta-analysis of literature data relating to the relationships between cadmium intake and toxicity indicators in humans. AB - The objective of the study was to determine the relationship between cadmium (Cd) intake and cadmium toxicity indicators by meta-analysis of literature data, in particular beta2-microglobulin (beta2MC), and to compare the results with the current Provisional Tolerable Weekly Intake (PTWI) set by FAO/WHO. The literature survey identified 79 feeding trials involving 27,537 people that were suitable for extraction of cadmium intake, levels in blood and urine and beta2 microglobulin in urine. There was an exponential increase in beta2-microglobulin with increases in cadmium intake above 302 micro g/day, which corresponds to a PTWI of 3.02 micro g/kg of body weight, when a safety margin of 10 is included. This compares with the current level set by FAO/WHO of 7 micro g/kg of body weight. Cadmium in blood and urine were also positively related to cadmium intake and participants' age. There were two principal components of variation in the data set, first: cadmium intake, concentrations of cadmium in blood, urine and beta2-microglobulin in urine, and second: duration and age of exposure. PMID- 17109841 TI - Effects of MK-801 on the expression of serine racemase and d-amino acid oxidase mRNAs and on the D-serine levels in rat brain. AB - We have investigated the acute effects of the increasing doses of non-competitive N-methyl-d-aspartate receptor antagonist MK-801 (0.2-1.6 mg/kg) on the expression of serine racemase and d-amino acid oxidase (DAO) mRNAs in several brain areas of rats. We have also evaluated the effects of the chronic administration of MK-801 (0.4 mg/kg) on the gene expression of serine racemase and DAO and on the d-serine concentrations. A dose-dependent augmentation of the expression of serine racemase mRNA was seen in most brain areas at both 1 and 4 h after the administration. In contrast, a drastic decline in the expression of DAO mRNA was observed in most brain areas 1 h after the MK-801 administration, whereas a dose dependent elevation in the expression of DAO mRNA was observed in most brain areas 4 h after the administration. The chronic MK-801 administration produced a significant increase in the expression of serine racemase mRNA in almost all brain areas, whereas no significant changes were found in the level of DAO mRNA in most brain areas. In addition, the chronic administration caused a slight but significant elevation in the concentrations of d-serine in the cortex and striatum. These present findings indicate that increasing the serine racemase mRNA and no changes in the DAO mRNA after the chronic administration could contribute to the elevation of the d-serine level in the forebrain, and that serine racemase and DAO could play an important role in the regulation of N methyl-d-aspartate receptors via the d-serine metabolism. PMID- 17109842 TI - Current understanding of the application of pyridinium oximes as cholinesterase reactivators in treatment of organophosphate poisoning. AB - This paper reviews the mechanisms of interaction of organophosphorus compounds with cholinesterases and clinical signs of acute poisoning. Further, we describe the current understanding of the mechanisms of action of pyridinium oximes pralidoxime (PAM-2), trimedoxime (TMB-4), obidoxime (LuH-6, Toxogonin), HI-6 and HLo-7 which are used as cholinesterase reactivators in the treatment of poisoning with organophosphorus compounds. We also review the most important literature data related to the efficacy of these oximes in the treatment of poisoning with warfare nerve agents soman, sarin, tabun, VX and cyclosarin and organophosphorus insecticides. Finally, we discuss the criteria for selection of oximes intended for further development as antidotes in poisoning with organophosphorus compounds and auto-injectors for their application in urgent situations. PMID- 17109843 TI - Neuroprotective activity of selective mGlu1 and mGlu5 antagonists in vitro and in vivo. AB - The neuroprotective potential of allosteric mGlu5 and mGlu1 antagonists such as 6 methyl-2-(phenylethynyl)-pyridin (MPEP)/[(2-methyl-1,3-thiazol-4 yl)ethynyl]pyridine (MTEP) and (3-ethyl-2-methyl-quinolin-6-yl)-(4-methoxy cyclohexyl)-methanone methanesulfonate (EMQMCM), was tested in vitro in organotypic hippocampal cultures and in the middle cerebral artery occlusion model of stroke in vivo. Both classes of agent have high selectivity toward mGlu sub-types and are active in animal models of various diseases indicating satisfactory CNS penetration. In organotypic hippocampal cultures MPEP showed high neuroprotective potency against sub-chronic (12 days) insult produced by 3 NP with an IC50 of c.a. 70 nM. In contrast, although the mGlu1 antagonist EMQMCM was also protective, it seems to be weaker yielding an IC50 of c.a. 1 microM. Similarly, in the transient (90 min) middle cerebral artery occlusion model of ischaemia in rats, MTEP seems to be more effective than EMQMCM. MTEP, at 2.5 mg/kg and at 5 mg/kg provided 50 and 70% neuroprotection if injected 2 h after the onset of ischaemia. At a dose of 5 mg/kg, significant (50%) neuroprotection was also seen if the treatment was delayed by 4 h. EMQMCM was not protective at 5 mg/kg (given 2 h after occlusion) but at 10 mg/kg 50% of neuroprotection was observed. The present data support stronger neuroprotective potential of mGlu5 than mGlu1 antagonists. PMID- 17109844 TI - Honokiol up-regulates prostacyclin synthease protein expression and inhibits endothelial cell apoptosis. AB - Honokiol is a bioactive compound extracted from the Chinese medicinal herb Magnolia officinalis. We recently demonstrated that honokiol inhibited arterial thrombosis through stimulation of prostacyclin (PGI2) generation and endothelial cell protection. The current study is designed to investigate its mechanism of stimulation of PGI2 generation and cell protection. 6-keto-PGF1alpha, the stable metabolite of PGI2, in the media of rat aortic endothelial cells was measured with radioimmunoassay kits. Indomethacin, an inhibitor of cyclooxygenase (COX) and tranylcypromine, a prostacyclin synthease inhibitor were used to ascertain the target enzyme affected by honokiol. Prostacyclin synthease protein levels in endothelial cells were determined by Western blot analysis using an anti-PGI2 synthease rabbit polyclonal antibody. Flow cytometry was used to quantify the apoptotic cells and spectrophotometry was used to test the caspase-3 activity. Honokiol (0.376-37.6 microM) increased the level of 6-keto-PGF1alpha in the media of normal endothelial cells. It counteracted the inhibitory effect of tranylcypromine on the PGI2 generation, but did not influence the effect of indomethacin; evidently, honokiol up-regulated the expression of prostacyclin synthease in the endothelial cells. These effects showed perfect concentration dependent behavior. In addition, at lower concentration (0.376-3.76 microM), honokiol significantly decreased the percentage of apoptotic endothelial cells induced by oxidized low-density lipoprotein (ox-LDL) and significantly lowered the activity of caspase-3 stimulated by ox-LDL. A high dose of honokiol (37.6 microM), however, failed to influence either of them. In conclusion, honokiol augments PGI2 generation in normal endothelial cells; its effect on PGI2 generation attributes to up-regulation of prostacyclin synthease expression; its cell protection may be correlated with its inhibition on apoptosis of endothelial cells. These findings have partly revealed the molecular mechanism of honokiol on inhibiting arterial thrombosis. PMID- 17109845 TI - Anti-inflammatory effects and mechanisms of the ethanol extract of Evodia rutaecarpa and its bioactive components on neutrophils and microglial cells. AB - Evodia rutaecarpa is commonly used as an anti-inflammatory drug in traditional Chinese medicine. We previously identified four bioactive compounds (dehydroevodiamine (I), evodiamine (II), rutaecarpine (III), and synephrine (IV)) from the ethanol extract of E. rutaecarpa, but their effects and mechanism(s) of action remain unclear. To study the anti-inflammatory potential and the possible underlying mechanism(s), their effects on phorbol-12-myristate-13-acetate (PMA)- and N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced reactive oxygen species production in neutrophils was studied, as well as lipopolysaccharide (LPS)-induced nitric oxide (NO) production and inducible NO synthetase (iNOS) expression in microglial cells. The ethanol extract of E. rutaecarpa displayed potent antioxidative effects against both PMA- and fMLP-induced reactive oxygen species production in neutrophils (with IC50 values of around 2.7-3.3 microg/ml). Although less potent than the ethanol extract of E. rutaecarpa, compounds I-IV all concentration-dependently inhibited PMA- and fMLP-induced reactive oxygen species production, with compound IV consistently being the most potent agent among these active components. The antioxidative effects of the ethanol extract of E. rutaecarpa and these compounds were partially due to inhibition (10%-33%) of NADPH oxidase activity, a predominant reactive oxygen species-producing enzyme in neutrophils, and to a minor extent to their direct radical-scavenging properties. The ethanol extract of E. rutaecarpa also inhibited LPS-induced NO production (with an IC50 of around 0.8 microg/ml) and iNOS upregulation in microglial cells that was partially mimicked by compounds I, II, and III, but not compound IV. Our results suggest that the ethanol extract of E. rutaecarpa and its four bioactive components all exhibited anti-inflammatory activities which could be partially explained by their different potentials for inhibiting NADPH oxidase-dependent reactive oxygen species and/or iNOS-dependent NO production in activated inflammatory cells. PMID- 17109846 TI - Agmatine and a cannabinoid agonist, WIN 55212-2, interact to produce a hypothermic synergy. AB - Agmatine blocks morphine withdrawal symptoms and enhances morphine analgesia in rats. Yet, the role of agmatine in the pharmacological effects of other abused drugs has not been investigated. The present study investigates the effect of agmatine administration on the hypothermic response to cannabinoids. Hypothermia is an effective endpoint because cannabinoid agonists produce a rapid, reproducible, and significant decrease in body temperature that is abolished by cannabinoid CB(1) receptor antagonists. WIN 55212-2, a cannabinoid agonist, was administered to rats by itself and with agmatine. WIN 55212-2 (1, 2.5, 5 and 10 mg/kg, i.m.) caused a significant hypothermia. Agmatine (10, 25 and 50 mg/kg, i.p.) was ineffective. For combined administration, agmatine (50 mg/kg, i.p.) enhanced the hypothermic effect of WIN 55212-2 (1, 2.5, 5 and 10 mg/kg, i.m.). The enhancement was strongly synergistic, indicated by a 2.7-fold increase in the relative potency of WIN 55212-2. The central administration of agmatine (25 and 50 mug/rat, i.c.v.) significantly increased the hypothermic effect of WIN 55212-2 (2.5 mg/kg, i.m.). This indicates that agmatine acts through a central mechanism to augment cannabinoid-evoked hypothermia. Idazoxan (2 mg/kg, i.p.), an imidazoline antagonist, blocked the enhancement by agmatine, thus suggesting that imidazoline receptor activation is required for agmatine to enhance cannabinoid evoked hypothermia. The present data reveal that agmatine and a cannabinoid agonist interact to produce a hypothermic synergy in rats. These results show that agmatine acts in the brain and via imidazoline receptors to enhance cannabinoid-evoked hypothermia. PMID- 17109847 TI - The anti-cancer drug-induced pica in rats is related to their clinical emetogenic potential. AB - Cancer chemotherapy is frequently accompanied by severe emesis. The anti-cancer drugs are classified according to their clinical emetogenic potential. We have already found that kaolin ingestion behavior "pica" is analogous to emesis in rats. The aim of this study was to examine the effects of the clinical emetogenic potential of anti-cancer drugs on the induction of the pica in rats. Rats were housed in individual cages with free access to food and kaolin pellets and the daily food and kaolin intakes were measured for 3 days after the intraperitoneal administration of anti-cancer drugs (cisplatin, cyclophosphamide, actinomycin D, 5-fluorouracil and vincristine). The drugs with high potential for inducing emesis, such as cisplatin and cyclophosphamide, induced pica in all animals on the day of administration and the behavior lasted during the observation period. The drugs with moderate emetogenic potential, i.e. actinomycin D and 5 fluorouracil, also induced pica on the first and second day after the drug administration but the kaolin intake was less than that of the drugs with high potential. Vincristine, a drug with low emetogenic potential, slightly increased the kaolin intake in rats on the only first day of the administration. Cyclophosphamide, actinomycin D and vincristine induced anorexia and decreased their body weight during the observation period. These results suggested that the both amounts of kaolin intake and duration of behavior in the anti-cancer drug induced pica are related to the clinical emetogenic potential of the drugs and the incidence of the anorexia is not related to their emetogenic potential. PMID- 17109848 TI - Influence of Crocetin on experimental atherosclerosis in hyperlipidamic-diet quails. AB - Antioxidants have been expected to have potential as antiatherogenic agents. Crocetin is a natural carotenoid antioxidant isolated from Gardenia jasminoids Ellis. Therefore, in the present study, we investigated the inhibitory effect of Crocetin on experimental atherosclerosis in quails. The atherosclerosis model was established by feeding hyperlipidamic diet to quail and Crocetin (25, 50, 100 mg/kg/day) was administered by oral gavage. At the 9th week, serum lipids, malondialdehyde and nitric oxide were measured, and Hematoxylin-Eosin (H&E) stains was used to investigate the histopathological changes of aorta. Results showed that Crocetin could reduce the levels of serum total cholesterol, triglyceride, low density lipoprotein cholesterol and inhibit the formation of aortic plaque. Crocetin could also reduce malondialdehyde and inhibit the descending of nitric oxide in serum. The results suggested that Crocetin could inhibit the formation of atherosclerosis in quails, which might be related to the hypolipidemic effects along with the antioxidative properties of Crocetin. PMID- 17109849 TI - Involvement of Ca2+ in the inhibition by crocetin of angiotensin II-induced ERK1/2 activation in vascular smooth muscle cells. AB - Crocetin, a carotenoid compound, was isolated from Gardenia jasminoids Ellis. Our recent study shows that crocetin inhibits angiotensin II-induced extracellular signal-regulated kinases 1/2 (ERK1/2) activation and subsequent proliferation in vascular smooth muscle cells (VSMCs). To further explore the mechanism involved, in the present study, we investigated the effect of Ca(2+) in the activation of ERK1/2 and whether Ca(2+) is involved in the suppression by crocetin of angiotensin II-induced ERK1/2 activation. Our findings showed that crocetin pretreatment partially attenuated both the intracellular Ca(2+) mobilization and the extracellular Ca(2+) influx induced by angiotensin II. Moreover, angiotensin II-induced ERK1/2 activation was completely abolished by acetoxymethyl ester of 1,2-bis(2-aminophenoxy)ethane-N,N,N ',N'-tetraacetic acid (BAPTA-AM), an intracellular Ca(2+) chelator, and partially inhibited by EGTA, an extracellular Ca(2+) chelator, or verapamil, an L-type Ca(2+) channel blocker. These findings suggest that Ca(2+) may play an important role in angiotensin II-induced ERK1/2 activation in VSMCs, and Ca(2+)-dependent pathway may be involved in the inhibitory effect by crocetin of angiotensin II-induced ERK1/2 activation. PMID- 17109850 TI - Antidepressant-like effects of novel triple reuptake inhibitors, PRC025 and PRC050. AB - Most currently prescribed antidepressants act by selectively increasing the synaptic availability of serotonin or norepinephrine, or through action on both serotonin and norepinephrine. However, most therapies require several weeks of treatment before improvement of symptoms is observed and not all patients respond to antidepressant treatment. One strategy that has emerged in new antidepressant development is the use of triple reuptake inhibitors, which inhibit reuptake of serotonin, norepinephrine, and dopamine. These compounds have been hypothesized to have a more rapid onset of activity and better efficacy over single or dual reuptake inhibitor antidepressants in part due to the addition of the dopamine component. We have developed novel compounds that are analogs of venlafaxine, of which two, racemic PRC025 ((2SR, 3RS)-N,N-dimethyl-3-cyclohexyl-3-hydroxy-2-(2' naphthyl)propylamine) and PRC050 ((2RS,3RS)-N-methyl-3-hydroxy-2-(2'-naphthyl)-3 phenylpropylamine), are highly potent at human serotonin, norepinephrine, and dopamine transporters and inhibit the reuptake of serotonin, norepinephrine, and dopamine into rat brain synaptosomes. These compounds were tested in animal models used to evaluate potential antidepressants: the forced swim test in rats and the tail suspension test in mice. In the forced swim test, both PRC025 and PRC050 reduced the time spent immobile and increased the time spent swimming, comparable to the effects seen with imipramine, a tricyclic antidepressant. In addition, both PRC025 and PRC050 were effective in reducing the time spent immobile in the tail suspension test, again with effects comparable to imipramine. Therefore it appears that our compounds may possess antidepressant activity and represent a new class of triple reuptake inhibitors. PMID- 17109851 TI - Pharmacological evidence that alpha2A- and alpha2C-adrenoceptors mediate the inhibition of cardioaccelerator sympathetic outflow in pithed rats. AB - It has been suggested that the alpha(2)-adrenoceptors mediating cardiac sympatho inhibition in pithed rats closely resemble the pharmacological profile of the alpha(2A)-adrenoceptor subtype. However, several lines of evidence suggest that more than one subtype may be involved. Thus, the present study has pharmacologically re-evaluated the receptor subtype(s) involved in the inhibitory effect of the alpha(2)-adrenoceptor agonist, B-HT 933, on the tachycardic responses elicited by selective cardiac sympathetic stimulation (0.03, 0.1, 0.3, 1 and 3 Hz) in desipramine-pretreated pithed rats. I.v. continuous infusions of B HT 933 (30 microg/kg min), which failed to modify the tachycardic responses to exogenous noradrenaline, inhibited those induced by preganglionic (C(7)-T(1)) stimulation of the cardiac sympathetic outflow at all frequencies of stimulation (0.03-3 Hz). This cardiac sympatho-inhibitory response to B-HT 933 was: (1) unaltered by saline (1 ml/kg) or the antagonists BRL44408 (100 microg/kg; alpha(2A)) or imiloxan (3000 and 10,000 microg/kg; alpha(2B)); (2) partially antagonized by BRL44408 (300 microg/kg) or MK912 (10 microg/kg; alpha(2C)) given separately; and (3) completely antagonized by rauwolscine (300 microg/kg; alpha(2)), MK912 (30 microg/kg) or the combination of BRL44408 (300 microg/kg) plus MK912 (10 microg/kg). Moreover, the above doses of antagonists, which are high enough to block their respective receptors, failed to block per se the tachycardic responses to sympathetic stimulation. These results suggest that the cardiac sympatho-inhibition induced by B-HT 933 in pithed rats is mainly mediated by stimulation of alpha(2A)- and alpha(2C)-adrenoceptors. PMID- 17109852 TI - The action of the novel gastrointestinal prokinetic prucalopride on the HERG K+ channel and the common T897 polymorph. AB - The human ether-a-go-go related gene (HERG) encodes the alpha-subunit of a delayed rectifier potassium channel important in the repolarisation of the cardiac action potential. Excessive action potential prolongation through HERG channel inhibition is associated with a risk of torsade de pointes arrhythmias and is a major challenge for drug development. The acute effects of the novel prokinetic prucalopride were examined on heterologously expressed HERG channels in human embryonic kidney (HEK) 293 cells using the whole-cell patch-clamp technique. Prucalopride inhibited HERG channels in a concentration-dependent manner with an IC(50) of 4.1 microM. Prucalopride significantly slowed channel deactivation and recovery from inactivation, accelerated and altered the extent of inactivation. Similar concentration-dependency and kinetic changes were observed with the minor T897 polymorphic HERG variant. Prucalopride block was frequency-independent due to rapid state-dependent block, with binding occurring in the open and inactivated states. Though prucalopride blocks HERG channels this is unlikely to be significant at clinically relevant concentrations. PMID- 17109853 TI - Inhibitory effect of 4-(2-aminoethyl)-benzenesulfonyl fluoride, a serine protease inhibitor, on PI3K inhibitor-induced CHOP expression. AB - Endoplasmic reticulum stress contributes to several diseases such as neurodegenerative disorders and diabetes. In the previous report, we found that phosphatidylinositol 3-kinase (PI3K) down-regulation is important for inducing CHOP expression, an endoplasmic reticulum stress-induced transcription factor. In the present study, we investigated the effect of 4-(2-aminoethyl)-benzenesulfonyl fluoride (AEBSF), a serine protease inhibitor, on PI3K inhibitor-induced CHOP expression. We found that AEBSF completely inhibited PI3K inhibitor-induced CHOP expression at both mRNA and protein levels. It is suggested that AEBSF is an important drug from a pharmacological point of view and the results may have important implications for understanding endoplasmic reticulum stress-related diseases. PMID- 17109854 TI - NO-1886, a lipoprotein lipase activator, attenuates vascular smooth muscle contraction in rat aorta. AB - The chemical compound [4-(4-bromo-2-cyano-phenylcarbamoyl)-benzyl]-phosphonic acid diethyl ester (NO-1886) is a lipoprotein lipase activator having beneficial effects on both diabetes control and the cardiovascular system. Preventing accumulation of lipids in the cell wall, in addition to improving insulin actions on vasculature, may indirectly contribute to the reducing effect of NO-1886 on vascular resistance. However, the direct effect of NO-1886 on vascular resistance, i.e., whether NO-1886 directly modulates the function of vascular endothelium and/or smooth muscle cells has not been investigated. In this study we therefore investigated the direct effect of NO-1886 on vascular contractility using rat aortic rings and cultured smooth muscle cell-line A10. The results show that administration of NO-1886 attenuated aortic contraction induced by phenylephrine and/or a high K(+) environment, in both the presence and absence of aortic endothelium. 1-(5-Chloronaphthalene-1-sulfonyl)homopiperazine hydrochloride (ML-9), a myosin light chain kinase (MLCK) inhibitor, blocked this inhibitory effect of NO-1886, whereas inhibitors of other signaling molecules such as calmodulin, protein kinase C and Rho-kinase had no effect. The vasorelaxant effect of NO-1886 was blocked in the absence of extracellular Ca(2+), or in the presence of the Ca(2+) channel inhibitor, verapamil. NO-1886 attenuated smooth muscle contraction induced by the cumulative addition of CaCl(2). In A10 cells, NO-1886 inhibited the membrane depolarization-induced initial peak of [Ca(2+)](i) in the presence of extracellular Ca(2+). This inhibition did not occur in the absence of extracellular Ca(2+). Taken together these results demonstrate that NO-1886 attenuates smooth muscle contraction and causes vasorelaxation by an extracellular Ca(2+)- and MLCK-dependent mechanism. PMID- 17109855 TI - Edaravone protects the vestibular periphery from free radical-induced toxicity in response to perilymphatic application of (+/-)-alpha-amino-3-hydroxy-5-methyl isoxazole-4-propionic acid. AB - Intracochlear infusion of (+/-)-alpha-amino-3-hydroxy-5-methyl-isoxazole-4 propionic acid (AMPA) was performed with a syringe pump in guinea pigs, and peripheral vestibular dysfunction was induced. Animals were administered edaravone systemically or topically. In the systemic application group, animals were administered edaravone once a day for 7 days after AMPA infusion. In the topical application group, edaravone-soaked gelfoam was placed on the round window membrane just after, 12 h after or 24 h after AMPA infusion. Spontaneous nystagmus was observed after AMPA infusion. Immunohistochemistry for 4-hydroxy-2 nonenal (4-HNE), a marker of free radical-induced lipid peroxidation, was performed 24 h after AMPA infusion. In addition, caloric tests were performed to evaluate vestibular function 1 week after AMPA infusion. Animals in both groups showed decreased spontaneous nystagmus, but results were not significant. Animals treated topically with edaravone within 12 h of AMPA infusion showed normal morphology of the ampullar sensory epithelia of the lateral semicircular canals and showed a good response to the caloric tests. 4-HNE immunoreactivity in the sensory epithelia was very low in these animals. In contrast, untreated animals and animals treated with edaravone systemically or topically 24 h after AMPA infusion showed morphologic hair cell damage, reduced caloric response and remarkable 4-HNE immunoreactivity in the sensory epithelia. These results indicate that topical application of edaravone within 12 h after damage protects the vestibular periphery from free radical-induced toxicity in response to intracochlear infusion of AMPA. PMID- 17109856 TI - Enhancement of the anti-immobility action of antidepressants by a selective 5-HT7 receptor antagonist in the forced swimming test in mice. AB - Using the forced swimming test in mice, we examined the effect of the following antidepressants: citalopram, imipramine, desipramine and moclobemide (which are characterized by different mechanisms of action), administered in combination with the selective 5-HT7 receptor antagonist (2R)-1-[(3-hydroxyphenyl)sulfonyl]-2 [2-(4-methyl-1-piperidinyl)ethyl]-pyrrolidine (SB 269970). All those drugs were given in doses which did not shorten the immobility time of mice. Citalopram (1.25 mg/kg), imipramine (10 mg/kg), desipramine (5 mg/kg) or moclobemide (10 mg/kg) administered jointly with SB 269970 (5 mg/kg), produced a significant antidepressant-like effect. None of the compounds studied, given alone or in combination, increased the spontaneous locomotor activity of mice. The obtained results indicate that blockade of 5-HT7 receptors may facilitate the anti immobility effect of antidepressants in mice. PMID- 17109857 TI - Fibulin-5 distribution in human eyes: relevance to age-related macular degeneration. AB - Fibulin-5 is an extracellular matrix glycoprotein that participates in elastogenesis. Mutations in the gene for fibulin-5 have been found to be associated with age-related macular degeneration. Little is known, however, about the expression of this gene in normal eyes or eyes with age-related macular degeneration. In this study, we evaluated the expression of the fibulin-5 protein in human donor eyes and localized this protein to Bruch's membrane and the intercapillary pillars of the choriocapillaris in normal eyes. In eyes with age related macular degeneration, fibulin-5 was localized to pathologic basal deposits beneath the retinal pigment epithelium as well as some small drusen. These results suggest that fibulin-5 may promote extracellular deposit formation in macular degeneration. PMID- 17109858 TI - Association of anti-mullerian hormone levels with obesity in late reproductive age women. AB - OBJECTIVE: To describe anti-mullerian hormone (AMH) levels in healthy late reproductive age women and test the hypothesis that AMH levels are lower in obese compared to non-obese women. DESIGN: Cross-sectional study of AMH levels. Longitudinal analysis of a subgroup with 10 AMH measures over 8 years to support the cross-sectional results. SETTING: A population-based cohort of healthy late reproductive-age women. PARTICIPANTS: Selected from the cohort to provide comparisons of body mass index (BMI), menopausal status, age and race (n = 122). INTERVENTIONS: AMH levels were determined from blood samples collected in the parent study. MAIN OUTCOME MEASURE: Serum levels of AMH. RESULTS: AMH levels were 65% lower in obese women compared to non-obese women (0.016 ng/mL and 0.046 ng/mL, respectively). The geometric mean ratio was 0.35; 95% CI 0.13, 0.92, P=.034. AMH levels were significantly lower in the menopausal transition compared to premenopausal women and were significantly lower in all age groups > or =40 years compared to the 35-39 year-old women. BMI remained significantly associated with AMH levels in multivariable models that included adjustments for menopausal status, age, race and cycle day. In the longitudinal analysis of a subgroup, obese women had significantly lower mean AMH levels over the 8-year interval compared to the non-obese women (0.459 ng/mL; CI 0.28, 0.75 and 0.566 ng/mL; CI 0.34, 0.94, respectively; P=.016), corroborating the cross-sectional study results. CONCLUSIONS: Obese women have lower AMH levels compared to non-obese women in the late reproductive years. The findings offer further evidence of the complex relationships between obesity and reproductive hormone levels in women. PMID- 17109859 TI - Randomized controlled trial comparing Thermachoice III* in the outpatient versus daycase setting. AB - OBJECTIVE: To evaluate outpatient versus daycase Thermachoice III endometrial ablation. DESIGN: Randomized, controlled trial. SETTING: Large United Kingdom teaching hospital. PATIENT(S): Seventy-three women with menorrhagia. INTERVENTION(S): In the outpatient cohort (n = 39), women took ibuprofen 800 mg the evening before, and 1 hour before, performing Thermachoice III. Intraoperatively, they were offered "rescue analgesia" in the form of inhaled nitrous oxide. Postoperatively, women were offered oral tramadol 50-100mg. Neither local anesthetic nor IV sedation was used. In the daycase cohort (n = 34), women underwent a general anesthetic, i.e., propofol and inhalation gases. Intraoperatively, intravenous fentanyl 100 microg and rectal diclofenac 100 mg were administered. MAIN OUTCOME MEASURE(S): Main outcome measures included overall discomfort following Thermachoice III; intraoperative pain scores and a need for rescue analgesia during outpatient Thermachoice III; nausea and vomiting rates; and total time in hospital and the need for an overnight stay. RESULT(S): Eighty-seven percent (n = 34) of women in the outpatient cohort completed the full 8-minute treatment. The remaining five women requested the procedure be stopped because of pain. The mean intraoperative pain scores for the outpatient cohort remained below 45 mm (range, 0-100 mm) for every stage of treatment. The majority (64%) of women from the outpatient cohort required no rescue analgesia whatsoever. There was no significant difference between the overall mean pain score following Thermachoice III for either the outpatient (59 mm; range, 0-100 mm) or daycase (53 mm; range, 0-100 mm) cohorts. However, women from both cohorts reported wide ranges of pain scores (0-100 mm). There was no significant difference between the Likert -style descriptions of overall discomfort between the two groups, with the majority of women rating the pain as none to moderate. The presence of severe dysmenorrhea was statistically significantly correlated with higher overall visual analogue-style pain scores and Likert-style descriptions of discomfort. Significantly fewer women in the outpatient group experienced nausea (13% versus 65% in the daycase group) or vomiting (0 versus 24% in the daycase group), or required postoperative antiemetics (0 versus 56% in the daycase group). The mean total time spent in hospital was significantly shorter for the outpatient cohort at 1 hour and 40 minutes, versus 8 hours 12 minutes for the daycase group. CONCLUSION(S): Outpatient Thermachoice III could be performed in the majority (87%) of women, and was associated with similar overall pain scores as daycase Thermachoice III, however, the range was wide. Outpatient Thermachoice III was associated with significantly less nausea, vomiting, need for antiemetics, and time spent in hospital than was the daycase procedure. PMID- 17109860 TI - Fluoroscopically guided synechiolysis for patients with Asherman's syndrome: menstrual and fertility outcomes. AB - OBJECTIVE: To describe a 5-year experience of fluoroscopically guided hysteroscopic synechiolysis for Asherman's syndrome. DESIGN: Retrospective, uncontrolled cohort study. SETTING: Department of Endo-Gynaecology, University of New South Wales, Royal Hospital for Women, Randwick, Sydney, New South Wales, Australia. PATIENT(S): Thirty consecutive patients with confirmed Asherman's syndrome (March 1999-March 2004). INTERVENTION(S): Women had hysteroscopy performed under general anesthetic, with the use of a spinal needle in parallel to the hysteroscope to perform synechiolysis. The progress of the procedure was determined by injecting radiographic contrast medium, and visualized with the use of an image intensifier. Individual procedures were terminated when the endometrial cavity was reconstructed, or at 60 minutes. Cyclic high-dose estrogen therapy was used to stimulate endometrial proliferation. Repeat procedures were performed monthly until the endometrial cavity was reestablished. MAIN OUTCOME MEASURE(S): Menstrual and fertility outcomes were obtained from patient records and by postal questionnaire. RESULT(S): Thirty patients were treated for Asherman's syndrome (13% AFS Grade I, 43% AFS Grade II, and 43% AFS Grade III). Prior to treatment, 60% of patients were amenorrheic. The median number of procedures per patient was 1.5 (range, 1-6), and the mean length of the procedure was 42 minutes (range, 10-70 minutes). After treatment, 96% had regular menses. Seventeen patients attempted to conceive after surgery, and 9 (53%) were successful. CONCLUSION(S): Hysteroscopic synechiolysis under image-intensifier control appears to be an effective treatment for Asherman's syndrome. PMID- 17109861 TI - Secondary abdominal compartment syndrome: a potential threat for all trauma clinicians. AB - Post-injury abdominal compartment syndrome (ACS) is an increasingly recognised phenomenon in critical care. During the last decade, ACS had also been characterised in patients without abdominal injuries, referred to as secondary ACS. Recent investigation has described this elusive syndrome better, with up to 70% mortality. Regardless of the cause of the syndrome and the nature of any extra-abdominal injuries, secondary ACS is invariably associated with massive fluid resuscitation. With a reliable, predictive model and new monitoring techniques, trauma clinicians should be able to identify the high-risk patient and attenuate the impact of this syndrome. PMID- 17109862 TI - The relationship between socio-demographic variables, job stressors, burnout, and hardy personality in nurses: an exploratory study. AB - BACKGROUND: Nursing is considered as a risk profession with high levels of stress and burnout, and these levels are probably increasing. OBJECTIVES: A model of prediction of burnout in nursing that includes socio-demographic variables, job stressors, and personal vulnerability, or resistance, is proposed. DESIGN: A cross-sectional correlational design was used. A sample of 473 nurses and student nurses in practice from three General Hospitals in Madrid (Spain) completed the "Nursing Burnout Scale". The data were analysed using descriptive statistics, Pearson correlations, and hierarchical multiple regression. RESULTS: The proposed model is a good predictor of the diverse burnout sub-dimensions: emotional exhaustion, depersonalisation, and lack of personal accomplishment. Significant predictors of burnout included age, job status, job stressors (workload, experience with pain and death, conflictive interaction, and role ambiguity), and hardy personality (commitment, control, and challenge). CONCLUSIONS: Identifying an integrative process of burnout among nurses is an essential step to develop effective managerial strategies so as to reduce the burnout problem. Specifically, the present study suggests that intervention aimed at reducing the risk for burnout may achieve better results if it includes enhancement of workers' hardy personality rather than just decreasing environmental stressors. PMID- 17109863 TI - Angiotensin II induces IL-6 expression and the Jak-STAT3 pathway in aortic adventitia of LDL receptor-deficient mice. AB - Angiotensin II (A-II), the major effector peptide of the renin angiotensin system potently accelerates progression of atherosclerosis. To investigate its effects on vascular inflammatory mechanisms, we elucidated vascular cytokine expression during early lesion development in A-II-infused atherosclerosis-prone LDLR-/- mice. Male LDLR-/- mice were placed on a "Western" high-fat diet for 4 weeks, followed by sham or A-II infusion for 7 weeks. Equal blood pressures and elevations in serum lipids were seen in both groups. Mice were sacrificed when significant A-II-induced plaque development was first detectable, aortae were explanted and culture media assayed for secreted cytokines. Nine cytokines were significantly induced with interleukin-6 (IL-6) being the most highly secreted. Local IL-6 production was confirmed by in situ mRNA hybridization and immunostaining, where the most abundant IL-6 was found in the aortic adventitia, with lesser production by the medial and intimal layers. Immunofluorescence colocalization showed IL-6 expression by fibroblasts and activated macrophages. Activation of downstream IL-6 signaling mediated by the Jak-STAT3 pathway was demonstrated by inducible phospho-Tyr705-STAT3 formation in the adventitia and endothelium (of IL-6+/+ mice only). These findings define cytokine profiles in the A-II infusion model and demonstrate that IL-6, produced by activated macrophages and fibroblasts in the adventitia, induces the Jak-STAT3 pathway during early A-II-induced atherosclerosis. PMID- 17109864 TI - Preanalytical sources of measurement error: the conundrum of the homocysteine hypothesis. PMID- 17109865 TI - The different effect of pioglitazone as compared to insulin on expression of hepatic and intestinal genes regulating post-prandial lipoproteins in diabetes. AB - This study investigates lipoprotein composition in diabetes before and after treatment with insulin or pioglitazone and its relationship to gene expression of five genes found in liver and intestine which are involved in cholesterol homeostasis. Thirty zucker diabetic fatty fa/fa and 10 lean rats were examined. mRNA for 3-hydroxy3-methylglutaryl coenzyme A reductase (HMGCoA), microsomal triglyceride transfer protein (MTTP), Niemann Pick C1-like 1 (NPC1L1) and ATP binding cassette transporters (ABC) G5 and G8 was determined using real-time, reverse transcriptase (RT-PCR). Cholesterol, triglyceride, apo B48 and apo B100 were elevated in chylomicrons and very low density lipoproteins (VLDL) of untreated diabetic animals (p<0.02). For similar blood glucose pioglitazone was more effective than insulin in normalising the lipoproteins. In diabetic animals, HMGCoA reductase, MTTP and NPC1L1 mRNA were significantly elevated (p<0.02) and ABCG5 and ABCG8 were significantly reduced (p<0.02) in the liver. Pioglitazone significantly reduced hepatic MTTP and NPC1L1 mRNA (p<0.0001) and significantly increased ABCG5 and G8 mRNA (p<0.0001) as compared to insulin. In conclusion diabetes was associated with major changes in mRNA levels of proteins involved in the regulation of post-prandial lipoproteins. Pioglitazone and insulin have different effects on post-prandial lipoprotein metabolism in part due their effect on genes regulating cholesterol synthesis and lipoprotein assembly. PMID- 17109866 TI - Effects of fenofibrate and simvastatin on HDL-related biomarkers in low-HDL patients. AB - The objective of the present study was to compare the effects of fenofibrate versus simvastatin on various HDL-related biomarkers in dyslipidemic patients with low HDL-C, in whom it is as yet unclear whether a statin or a fibrate is the most appropriate treatment. Fifty-two patients received either fenofibrate (160 mg/day) or simvastatin (40 mg/day) for 8 weeks in a randomized, double-blind, parallel group trial. Simvastatin effectively lowered plasma LDL-C and apoB levels, but did not change plasma HDL levels and HDL-related biomarkers, except for a small, significant increase in the capacity of plasma to promote SR-BI mediated cholesterol efflux. Fenofibrate did not affect plasma LDL-C levels but lowered triglycerides, and exerted a remarkable HDL-C raising activity (+22%), with patients in the lowest range of HDL-C getting the maximal benefit. The HDL-C raise was associated with a shift of HDL from large to small particles, and from LpA-I to LpA-I:A-II, which might explain the observed increase in the plasma capacity to promote ABCA1 mediated efflux with no changes in SR-BI efflux. The distinct and complementary effects of fenofibrate and simvastatin on lipid parameters and HDL-related biomarkers suggest that a combination therapy with the two drugs in dyslipidemic patients with low HDL would be fully justified. PMID- 17109867 TI - The influence of salt type on the retention of bovine serum albumin in ion exchange chromatography. AB - In this paper, an analysis of the influence of the salt types, NaCl, NaCH3COO, Na2SO4 and Na3C6H5O7, on the isocratic retention behaviour of bovine serum albumin (BSA) on two anion-exchangers media (Source 30Q and TSK Gel Super Q 5 PW) has been presented. The retention data demonstrated that the mechanism of protein retention in ion-exchange chromatography (IEC) involves interactions between the protein solute, the mobile phase constituents and the stationary phase. The effect of protein activity coefficient in the mobile phase on the protein retention volumes is verified. PMID- 17109868 TI - Determining the composition and microstructure of ethylene-propylene copolymers by pyrolysis-gas chromatography. AB - The average composition and detailed microstructure of copolymers of ethylene and propylene have been studied by pyrolysis-gas chromatography (Py-GC), using a statistical modeling approach to analyze the data. The trimer distribution obtained from Py-GC is used to infer monomer arrangement information, which is quantified in terms of a number-average sequence length for each monomer. These values are used to define the microstructure and to calculate the average composition. Compared with other available techniques, Py-GC provides a simple, quick and reliable approach to study the microstructure and composition of polyolefin copolymers. Details of this Py-GC method are discussed, including an examination of its advantages and disadvantages, and a summary of the qualitative and quantitative analysis aspects of this approach is presented. The combination of a statistical modeling approach with Py-GC to study copolymer composition and microstructure allows one to investigate the complex problem of monomer arrangement in copolymers using a widely available analytical technique. We expect that with further advances in separation technology, especially two dimensional gas chromatography (GC x GC), research of this type will be become increasingly accurate and reproducible in the near future. PMID- 17109869 TI - Comparative study of two chromatographic methods for quantifying 2,4,6 trichloranisole in wines. AB - Here we present the validation and the comparative study of two chromatographic methods for quantifying 2,4,6-trichloroanisole (TCA) in wines (red, rose and white wines). The first method involves headspace solid-phase microextraction and gas chromatography with electron-capture detection (ECD). The evaluation of the performance parameters shows limit of detection of 0.3 ng l(-1), limit of quantification of 1.0 ng l(-1), recoveries around 100% and repeatability of 10%. The second one implies a headspace solid-phase microextraction and gas chromatography with mass spectrometric detection. The performance parameters of this second method are limit of detection of 0.2 ng l(-1), limit of quantification of 0.8 ng l(-1) and repeatability of 10.1%. From the comparative study we can state that both methods provide similar results and the differences between them are the better sensitivity of the GC-ECD method and the very shorter chromatogram running time of the GC-MS method. The two methods are able to quantify TCA below the sensorial threshold in red, rose and white wines using just a calibration graph, thus they could be a very good tool for quality control in wineries. PMID- 17109870 TI - Determination of hydroxycinnamic acids and volatile phenols in wort and beer by isocratic high-performance liquid chromatography using electrochemical detection. AB - The suitability of a simple and rapid isocratic RP-HPLC method with amperometric electrochemical detection for the simultaneous detection and quantification of hydroxycinnamic acids and their corresponding aroma-active volatile phenols in wort and beer is reported. The technique gives good specificity and sensitivity, and can therefore be used for routine monitoring of hydroxycinnamic acids in wort and the development of volatile phenolic flavour compounds during the beer production process and subsequent conservation. PMID- 17109871 TI - Content uniformity and assay requirements in current regulations. AB - The acceptance of a tablet batch is based both on the content uniformity test and on the assay. It is shown that these two characteristics are not independent, and the acceptance criteria for them are not even consistent. For content uniformity range three methods of calculation are compared: the present European Pharmacopoeia method, a tolerance range method with improved k tolerance factor and a one-way random effects analysis of variance model. To resolve the inconsistency several options are discussed: applying the holistic content uniformity range alone; using content uniformity standard deviation and assay mean simultaneously or applying a criterion based on Taguchi's quadratic loss function. PMID- 17109872 TI - Selective pressurized liquid extraction of polychlorinated dibenzo-p-dioxins, dibenzofurans and dioxin-like polychlorinated biphenyls from food and feed samples. AB - Selective pressurized liquid extraction (PLE) of polychlorinated dibenzo-p dioxins/dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl PCBs) from various food and feed samples was performed with a selective PLE method previously developed for bulk PCBs. The method utilizes sulfuric acid impregnated silica inside the extraction cell to oxidize coextracted fat. Extractions were performed at 100 degrees C with n-heptane for 5 min in two cycles. Data obtained by selective PLE combined with gas chromatography/high resolution mass spectrometry (GC-HRMS) were compared to concentrations derived from reference laboratories applying conventional sample preparation and GC-HRMS. Experiments performed on spiked vegetable oil, naturally contaminated crude fish oil and oil containing compound feed samples showed good results for these relatively simple matrices. The accuracy was generally +/-20% as compared to spiked levels or to values obtained by the reference laboratories. The precision, measured as the relative standard deviation (RSD) for 2,3,7,8-tetrachlorodibenzo p-dioxin toxic equivalency values (TEQs), was below 10% in all cases. The method was also tested on naturally contaminated herring tissue, chicken tissue, pork tissue and sepiolitic clay, which all caused some trouble. It was observed that sufficient amounts of sodium sulfate should be used for dehydration of tissue samples and additionally, the cells should not be packed too dense in order to avoid suppressed extraction efficiency. Once this was attended to, satisfactory data could be obtained, except for sepiolithic clay. This study demonstrates that selective PLE can be applied with success to a number of food and feed matrices in analysis of PCDD/Fs and dl-PCBs. Since the fat removal step is on-line, the selective PLE method will reduce time and solvent consumption for sample preparation as compared to traditional clean-up. PMID- 17109873 TI - A method for measuring semi- and non-volatile organic halogens by combustion ion chromatography. AB - Recent studies have shown that various semi- and non-volatile organohalogen compounds are ubiquitous in the environment: these include halogenated dioxins including chlorinated dioxins, other persistent organic pollutants (POPs), brominated flame retardants (BFRs), and perfluorooctane sulfonate (PFOS). However, monitoring and assessment of these compounds by the analyses of individual compounds and their isomers is onerous because of their low environmental concentrations and large number of compounds. In this study, we have developed a new method that is capable of screening and monitoring an array of organohalogen compounds efficiently by combustion ion chromatography (CIC) - the new analyzer that serially connects combustion furnace and ion chromatograph. Analyzer performance was evaluated in terms of its applicability, reproducibility, and sensitivity as limit of detection (LOD). Recoveries of organochlorine, organobromine, and organoiodine compounds by the CIC were between 97 and 105%; those of organofluorine compounds were from 86 to 91%. In all cases, the relative standard deviation of five analyses was 4% or smaller. The analyzer would exhibit good sensitivity for various environmental matrices (e.g., 2.8-31ng X/g-soil, 1.4-16ng-X/L-water, and 9.2-100ng-X/m3N-gas). The method is fast and can provide information regarding the occurrence of organohalogen compounds within 1 or 2 days after sampling. Applicability of the new method for the assessment of contamination in flue gas and fly ash was also demonstrated. Our results show that the method is efficient to investigate emission sources and areas contaminated by organohalogen compounds. PMID- 17109874 TI - Fibre selection based on an overall analytical feature comparison for the solid phase microextraction of trihalomethanes from drinking water. AB - This paper describes the optimization of solid-phase microextraction (SPME) conditions for three different fibres (Carboxen-polydimethylsiloxane (CAR-PDMS), divinylbenzene-Carboxen-polydimethylsiloxane (DVB-CAR-PDMS) and polydimethylsiloxane-divinylbenzene (PDMS-DVB)) used to determine trihalomethanes (THMs) in water by headspace solid-phase microextraction and gas chromatography (HS-SPME-GC). The influence of temperature and salting-out effect was examined using a central composite design for each fibre. Extraction time was studied separately at the optimum values found for temperature and sodium chloride concentration (40 degrees C and 0.36g mL-1). The HS-SPME-GC-MS method for each fibre was characterised in terms of linearity, detection (LOD) and quantification (LOQ) limits and repeatability. The fibre PDMS-DVB was selected as it provided a broader linear range, better repeatability and lower detection and quantification limits than the others, particularly CAR-PDMS fibre. The accuracy of the proposed method using the PDMS-DVB fibre was checked by a recovery study in both ultrapure and tap water. A blank analysis study showed the absence of memory effects for this fibre. The reproducibility (expressed as a percentage of relative standard deviation) was 6-11% and the detection limits were between 0.078 and 0.52microgL 1 for bromoform and chloroform, respectively. Finally, the method was applied to determine THM concentration in two drinking water samples. PMID- 17109875 TI - Drop manipulation and surgery using electric fields. AB - We study the dynamics of a slender drop sandwiched between two electrodes using lubrication theory. A coupled system of evolution equations for the film thickness and interfacial charge density is derived and simplified for the case of a highly conducting fluid. The contact line singularity is relieved by postulating the existence of a wetting precursor film, which is stabilised by intermolecular forces. We examine the motion of the drop as a function of system parameters: the electrode separation, beta, an electric capillary number, C, and a spatio-temporally varying bottom electrode potential. The possibility of drop manipulation and surgery, which include drop spreading, translation, splitting and recombination, is demonstrated using appropriate tuning of the properties of the bottom potential; these results could have potential implications for drop manipulation schemes in various microfluidic applications. For relatively small beta and/or large C values, the drop assumes cone-like structures as it approaches the top electrode; the latter stages of this approach are found to be self-similar and a power-law exponent has been extracted for this case. PMID- 17109876 TI - Turbulent hydrodynamic stress induced dispersion and fragmentation of nanoscale agglomerates. AB - High pressure dispersion nozzles of 2.5-10 mm length and 125 microm diameter have been characterized in terms of fluid dynamics and dispersion experiments at 100 1400 bar. Elongational stresses at the nozzle entry (5 x 10(5) Pa) and turbulent stresses up to 10(5) Pa at a Reynolds number of 25,000 in turbulent channel flow are identified crucial for desagglomeration and aggregate fragmentation. Maximum stresses are calculated on representative particle tracks and related to agglomerate breakage. Agglomerates in the experimental study are in the range of the Kolmogorov micro scale (100-400 nm) and therefore break due to turbulent energy dissipation in viscous flow. Bond strength distributions could be determined experimentally from particle size distributions and fluid dynamics simulations, with primary particle erosion determined as dispersion mechanism for diffusion flame silica particles. Nanoscale agglomerates show a power law scaling for breakage with scaling exponents diverging from theory of floc dispersion. This is attributed to their strong bonding by sinter necks. PMID- 17109877 TI - Bromide and iodide removal from waters under dynamic conditions by Ag-doped aerogels. AB - The objective of this study was to analyze the efficiency of Ag-doped aerogels in the removal of bromide and iodide from water. To test the applicability of these aerogels in water treatment, adsorption of bromide and iodide was studied under dynamic conditions using waters from Lake Zurich and a mineral water. The results obtained by using these waters showed a high breakthrough volume (V(0.02)=0.4 L) of the columns, while the height of the mass transfer zone (H(MTZ)=6.8 cm) was low, regardless of the anion under study. Bromide- and iodide-saturated columns were regenerated with NH4OH. No change in the column characteristics was observed after two regeneration treatments, regardless of the type of water considered. PMID- 17109878 TI - Wetting and electrical properties of the human hair surface: delipidation observed at the nanoscale. AB - The electrostatic properties and the wetting behaviour of the human hair surface at the nanometric scale have been investigated by using atomic force microscopy (AFM). Surface potential imaging was used to determine the electrostatic properties while non-contact mode AFM was used to investigate the wetting properties of a test liquid, squalane. We have studied natural hair and hair in which different covalently (18-methyleicosanoic acid) and non-covalently bound fatty acids present at the cuticle surface were selectively extracted. This study shows how the removal of these acids causes various profound changes in hair wettability at the cuticle scale. PMID- 17109879 TI - Children's engagement and competence in personal recollection: effects of parents' reminiscing goals. AB - Parents' goal orientations in parent-child reminiscing were examined in this study, where 28 preschoolers (mean age = 46 months) experienced a standardized event. Dyads discussed the event that evening, with parents randomly assigned to either an "outcome-oriented" or a "process-oriented" condition. Outcome-oriented parents, who were told that children subsequently would be tested on event related recall, were more controlling in these conversations compared with process-oriented parents, who were told that children's personal perspective would be assessed. Parents did not differ in their provision of structure. Children were interviewed 2 weeks later. Autonomy support in the parent-child conversation predicted children's engagement in the interview. Parental structure predicted children's recall of details and the coherence of their memories. Effects of parental reminiscing styles for children's memory and motivation to reminisce are discussed. PMID- 17109880 TI - A novel approach using streptavidin magnetic bead-sorted in vivo biotinylated survivin for monoclonal antibody production. AB - One major obstacle in antibody production is the lack of highly purified immunogen. In this study, we describe an alternative strategy to circumvent this problem. A nucleotide sequence encoding a full-length of human survivin was cloned into pAK400cb. After transforming into an E. coli Origami B strain, survivin-biotin carboxyl carrier protein (BCCP) fusion protein was generated in the cytoplasm, where the BCCP domain serves as a target for in vivo biotinylation. The biotinylated heterologous protein was subsequently immobilized on streptavidin-coated magnetic particles and separated from other proteins in a magnetic field. The survivin-coated beads were used to raise immune responses in BALB/c mice for hybridoma production. A number of hybrid clones were found to secrete anti-survivin antibodies. Three established clones were selected for single cell cloning. All generated monoclonal antibodies specifically reacted with the standard human recombinant survivin. Two out of three monoclonal antibodies recognized survivin in tumor extracts. The present method has advantages in facilitating monoclonal antibody production by making antigen purification steps unnecessary. PMID- 17109881 TI - Actions and therapeutic potential of G-CSF and GM-CSF in cardiovascular disease. AB - Despite their names, the cytokines granulocyte- and granulocyte-macrophage-colony stimulating factor (G-CSF and GM-CSF respectively) have actions far beyond simply stimulating the proliferation of neutrophil and monocyte lineage cells. A comprehensive body of evidence now exists demonstrating that G-CSF and GM-CSF effectively mobilize bone-marrow-derived progenitor cells into the peripheral circulation. These mobilized progenitor cells can be conveniently harvested for use in reconstituting bone marrow by transplantation after myelo-ablative treatment of hematological malignancies. In addition, much evidence has recently emerged to suggest that these cytokines may have multiple direct and indirect beneficial cardiovascular effects--including neovascularization of ischemic myocardium and reducing the extent of myocardial damage after infarction. Based on this knowledge and a strong safety record in hematological applications, a number of early clinical trials have evaluated the use of G-CSF or GM-CSF in patients with both acute and chronic myocardial ischemia. Although the interpretation of these trials is complicated by heterogeneity in study design, small patient numbers and methodological concerns related to appropriate selection and blinding of patients, the results of ongoing larger phase II/III trials should soon be available to determine if these agents will be useful additions to the cardiovascular armamentarium. PMID- 17109882 TI - The mechanism of discrimination between cognate and non-specific DNA by dimeric b/HLH/LZ transcription factors. AB - The Myc/Max/Mad proteins are basic region-helix-loop-helix-leucine zipper (b/HLH/LZ) transcription factors that regulate the transcription of numerous genes involved in cell growth and proliferation. The Max protein is the obligate heterodimeric partner of the Myc and Mad proteins. Heterodimerization and DNA binding to target gene promoters are mediated by the b/HLH/LZ domains. Max can also form a homodimeric b/HLH/LZ. The enhanced expression of Myc and binding to promoters of target genes contribute to almost every aspect of tumor biology. However, the detailed mechanism by which dimeric and heterodimeric b/HLH/LZs discriminate cognate DNA (E-Box: CACGTG) from non-specific sequences in the target gene promoters is still unknown. Here, we use the Max b/HLH/LZ homodimer as a model for this class of transcription factors in the characterization and understanding of the mechanism of discrimination between the E-Box and non specific DNA sequences. We report the characterization of a cognate and a non specific Max b/HLH/LZ/DNA complex by EMSA, CD and NMR. Our results support a detailed mechanism by which dimeric b/HLH/LZ transcription factors can discriminate E-Box sequences from non-specific DNA. The mechanism proceeds via the conformational selection of fitting b/HLH/LZ homodimers with the basic region only partially helical. Next, the basic region undergoes a DNA-assisted folding or induced-fit. It is this step that provides the discrimination by stabilizing and destabilizing the alpha-helical conformation of the basic region in the cognate and non-specific complexes, respectively. This leads to a low affinity complex with a higher probability of being dissociated and hence to discrimination. A description of the side-chains and nucleotides proposed to be involved in the discrimination process is provided. PMID- 17109883 TI - The folding pathway of T4 lysozyme: the high-resolution structure and folding of a hidden intermediate. AB - Folding intermediates have been detected and characterized for many proteins. However, their structures at atomic resolution have only been determined for two small single domain proteins: Rd-apocytochrome b(562) and engrailed homeo domain. T4 lysozyme has two easily distinguishable but energetically coupled domains: the N and C-terminal domains. An early native-state hydrogen exchange experiment identified an intermediate with the C-terminal domain folded and the N-terminal domain unfolded. We have used a native-state hydrogen exchange-directed protein engineering approach to populate this intermediate and demonstrated that it is on the folding pathway and exists after the rate-limiting step. Here, we determined its high-resolution structure and the backbone dynamics by multi-dimensional NMR methods. We also characterized the folding behavior of the intermediate using stopped-flow fluorescence, protein engineering, and native-state hydrogen exchange. Unlike the folding intermediates of the two single-domain proteins, which have many non-native side-chain interactions, the structure of the hidden folding intermediate of T4 lysozyme is largely native-like. It folds like many small single domain proteins. These results have implications for understanding the folding mechanism and evolution of multi-domain proteins. PMID- 17109884 TI - A deletion variant study of the functional role of the Salmonella flagellin hypervariable domain region in motility. AB - The eubacterial flagellum is a complex structure with an elongated extracellular filament that is composed primarily of many subunits of a flagellin protein. The highly conserved N and C termini of flagellin are important in its export and self-assembly, whereas the middle sequence region varies greatly in size and composition in different species and is known to be deletion-tolerant. In Salmonella typhimurium phase 1 flagellin, this "hypervariable" region encodes two solvent-exposed domains, D2 and D3, that form a knob-like feature on flagella fibers. The functional role of this structural feature in motility remains unclear. We investigated the structural and physiological role of the hypervariable region in flagella assembly, stability and cellular motility. A library of random internal deletion variants of S. typhimurium flagellin was constructed and screened for functional variants using a swarming agar motility assay. The relative cellular motility and propulsive force of ten representative variants were determined in semi-solid and liquid medium using colony swarming motility assays, video microscopy and optical trapping of single cells. All ten variants exhibited diminished motility, with varying extents of motility observed for internal deletions less than 75 residues and nearly complete loss of motility for deletions greater than 100 residues. The mechanical stability of the variant flagella fibers also decreased with increasing size of deletion. Comparison of the variant sequences with the wild-type sequence and structure indicated that all deletions involved loss of hydrophobic core residues, and removal of both partial and complete segments of secondary structure in the D2 and D3 domains. Homology modeling predicted disruptions of secondary structures in each variant. The hypervariable region D2 and D3 domains appear to stabilize the folded conformation of the flagellin protein and contribute to the mechanical stability and propulsive force of the flagella fibers. PMID- 17109885 TI - Solution structure of Escherichia coli PapI, a key regulator of the pap pili phase variation. AB - Pyelonephritis-associated pili (pap) allow uropathogenic Escherichia coli to bind to epithelial cells and play an important role in urinary tract infection. Expression of pap is controlled by a phase-variation mechanism, based on the two distinct heritable states that are the result of adenine N6-methylation in either of the two GATC sequences in its regulatory region. The methylation status of these two sequences is sensed by the action of two proteins, Lrp and PapI, and they play a central role in determining pap gene expression in both phase-ON and phase-OFF cells. We used modern NMR techniques to determine the solution structure and backbone dynamics of PapI. We found its overall fold resembles closely that of the winged helix-turn-helix family of DNA-binding proteins. We determined that PapI possesses its own DNA-binding activity, albeit non-sequence specific, independent of Lrp. PapI appears to bind to DNA with a K(d) in the 10 microM range. Possible mechanisms by which PapI might participate in the regulation of the pap operon are discussed in light of these new findings. PMID- 17109886 TI - Beta-catenin signaling in fibroproliferative disease. AB - BACKGROUND: Beta-catenin has been historically recognized as both an intermediate in the "canonical Wnt signaling pathway" and as a component of functional adherens junctions. MATERIALS AND METHODS: Cellular accumulation of beta-catenin levels can result in transactivation of gene transcription and cellular proliferation during normal cellular and disease development. Recent evidence has identified beta-catenin in an additional role as a component of cutaneous wound healing. RESULTS: This finding is in keeping with previous observations that post translational modifications of beta-catenin that are associated with its cytoplasmic accumulation are frequently observed in fibroproliferative diseases with characteristics of dysregulated wound healing. These diseases include hypertrophic scar formation, aggressive fibromatoses, Lederhose disease, and Dupuytren's contracture (DC). CONCLUSIONS: While its precise roles in disease initiation and progression remain to be explored, this review highlights our current knowledge of beta-catenin regulation and describes some potential upstream mediators of beta-catenin accumulation and signaling in fibroproliferative disease. PMID- 17109887 TI - Rapamycin inhibits proliferation of estrogen-receptor-positive breast cancer cells. AB - BACKGROUND: Estrogen-receptor (ER)-positive breast cancers comprise the majority of sporadic breast cancers. Although 50% respond to antihormonal treatment, both primary and acquired resistance limit the utility of this therapy, and other agents are needed. Rapamycin, an inhibitor of the mammalian Target of Rapamycin (mTOR), possesses antitumor activity against many tumors including breast tumors, and particularly against ER-positive breast cancer cell lines. The sensitivity of these cells to rapamycin has been attributed to activation of the PI3K/Akt/mTOR pathway by nongenomic ER signaling. The purpose of this study was to evaluate the efficacy of rapamycin against ER-positive breast cancer, particularly under 17beta-estradiol (E2)-dependent conditions, and to investigate mechanisms of rapamycin-sensitivity in ER-positive cells. MATERIALS AND METHODS: Breast cancer cell lines were tested for sensitivity to rapamycin. Antiproliferative effects of rapamycin, alone and in combination with tamoxifen, were assessed under E2 dependent conditions. Western blot analysis was used to detect activation of mTOR by nongenomic ER signaling. RESULTS: Rapamycin effectively inhibits proliferation of the ER-positive MCF-7 cell line. In our system, this sensitivity is probably not due to nongenomic ER activation of the PI3K/Akt/mTOR pathway; rapid stimulation of mTOR occurred nonspecifically after medium replacement, and addition of E2 stimulated mTOR only after 1 h. Combining rapamycin and tamoxifen under E2-dependent conditions yielded additive/synergistic effects at effective concentrations. CONCLUSIONS: These results suggest that rapamycin may be an effective treatment for ER-positive breast cancer, either alone or in combination with tamoxifen, and also may be a potential therapy for tamoxifen-resistant cancers. PMID- 17109888 TI - Racial disparities in the management of pediatric appenciditis. AB - BACKGROUND: Our objective was to compare the racial differences in incidence and management of pediatric appendicitis. MATERIALS AND METHODS: Data for this study come from two large national hospital discharge databases from the Agency for Healthcare Research and Quality Healthcare Costs and Utilization Project: The Nationwide Inpatient Sample (NIS) and the Kids' Inpatient Database (KID). Analysis was restricted to age less than 18 years with an ICD-9 diagnosis of either simple (540.9) or complex (540.0 and 540.1) appendicitis. Data were weighted to represent national estimates. Incidence was defined as the number of new disease cases divided by the number of at risk hospitalized children. RESULTS: The data for this study contained an estimated 428,463 [95% confidence interval (CI) = 414, 672-442, 253] cases of appendicitis, representing approximately 65,000 to 75,000 cases annually. Multi-variant analysis suggests that African-Americans, as compared to Caucasians, were less prone to develop appendicitis [odds ratio (OR) = 0.39, 95% CI (0.38, 0.41)], but less frequently underwent laparoscopic treatment [OR = 0.78, 95% CI (0.74, 0.87)], and were more likely to have complex appendicitis [OR = 1.39, 95% CI (1.30, 1.49)]. In contrast, Hispanics were more likely than Caucasians to both develop appendicitis [OR = 1.48, 95% CI (1.41, 1.56)] and to have complex disease [OR = 1.10, 95% CI (1.05, 1.16)]. The incidence of appendicitis was less frequent in females versus males [OR = 0.69, 95% CI (0.68, 0.70)] but the likelihood of laparoscopic exploration was higher [OR = 1.39, 95% CI (1.34, 1.43)]. Finally, children with public insurance [OR = 1.25, 95% CI (1.21, 1.29)] and uninsured children [OR = 1.10, 95% CI (1.04, 1.16)] were more likely to have complex appendicitis when compared to children with private insurance. CONCLUSIONS: African-American children with appendicitis have lower overall hospitalization rates, higher rates of perforation, a greater delay to surgical management, and lower laparoscopic rates. In contrast, Hispanic children more frequently had appendicitis and complex disease. The treatment of African-American and Hispanic children overall was associated with a longer hospital stay and higher charges. The lower incidence of appendicitis in African-American children is incompletely understood and the disparity in surgical management among minority children remains troubling. PMID- 17109889 TI - Interleukin-1beta and interleukin-6 stimulate matrix metalloproteinase-9 secretion in cultured myenteric glia. AB - Western blotting of culture media of myenteric glia stained positive for GFAP revealed increased secretion of matrix metalloproteinase-9 (MMP-9) in interleukin 1beta (IL-1beta) stimulated cells (10 ng/mL) versus control (142 +/- 19 versus 42 +/- 19, P < 0.05). Interleukin-6 (IL-6) stimulated cells also showed increased expression of MMP-9 (10 ng/mL) versus control (69 +/- 14 versus 4 +/- 2, P < 0.01). Control and cytokine-stimulated cells secreted MMP-2 constituitively. Gelatin zymography demonstrated that products were biologically active. Cytoplasmic staining for MMP-9 was detected in IL-1beta and IL-6-stimulated cells but was negligible in controls. Cultured myenteric glia are responsive to IL 1beta and IL-6 stimulation by secreting MMPs into the extracellular environment. PMID- 17109890 TI - The STAT4 and STAT6 pathways in pancreatitis-associated lung injury. AB - BACKGROUND: The STAT pathways are integral to the inflammatory response and these proteins provide a direct link between the cytokine receptors and cytokine induced gene transcription. We examined the roles of STAT4 and STAT6 in lung injury after caerulein-induced severe acute pancreatitis. We hypothesized that a modified organ expression of cytokines and chemokines that occurs in transgenic mice may affect the systemic response to severe acute pancreatitis. METHODS: Acute pancreatitis [13-hourly intraperitoneal injections of caerulein (50 microg/kg body weight, 0.2 mL) or the same volume of saline] was induced in wild type (BALB/c) and transgenic (STAT4 or STAT6) mice of the same background, 7 to 8 weeks old. The pancreatic and lung tissues were collected at 1, 6, 12, and 24 h after the completion of caerulein administration. Tissue leukocyte sequestration was assessed by myeloperoxidase (MPO) activity. Standard histological staining hematoxylin and eosin was performed and blindly scored by a pathologist for evidence of lung injury (pulmonary edema, accumulations of neutrophils and mononuclear cells, thickness of alveolar-capillary membrane, perivascular infiltrate, and hemorrhage). RESULTS: Caerulein-treated wild-type mice exhibited increased lung injury score at 1 through 12 h, as compared to saline controls. As compared to wild-type, STAT6-deficient mice had increased lung injury from 1 to 6 h, with full recovery by 12 h. An opposite pattern was observed in STAT4 deficient mice with mild injury seen at 1 and 6 h, and maximal injury at 12 h. MPO activity was significantly increased at 6 h in caerulein-treated wild-type mice compared to saline-treated controls. Caerulein-treated STAT6 and STAT4 mice had markedly increased MPO activity as compared with their saline controls during the first 6 h. Both caerulein-treated STAT4- and STAT6-deficient mice had significantly increased MPO activity in comparison with wild-type mice with pancreatitis at 6 h. CONCLUSION: We found the maximal lung injury after caerulein induced pancreatitis occurred at different time-points in STAT4 and STAT6 deficient mice. These temporal differences may suggest alternative roles in the systemic inflammatory response associated with pancreatitis. PMID- 17109891 TI - Influence of intraperitoneal application of taurolidine/heparin on expression of adhesion molecules and colon cancer in rats undergoing laparoscopy. AB - BACKGROUND: Recent experimental studies have shown that intraperitoneal administration of taurolidine/heparin causes a reduction of local tumor growth after laparoscopy in rat models. It might be that the anti-adherent activities of these agents are responsible for this effect. In this study we investigated the adhesion molecules E-cadherin, beta1-integrin, and CD44. MATERIALS AND METHODS: Following a 10,000 colon adenocarcinoma cells' (DHD/K12/TRb) intraperitoneal application a cecum resection and a partial parietal peritoneum resection (1 x 1 cm) were performed using a three trocar technique in 30 BD IX rats. After randomization in two groups, the cecum suture line and the parietal peritoneal defect were either lavaged with 1 mL of 0.5% taurolidine/10 IU heparin or with equal amounts of 0.9% normal saline solution. Rats were sacrificed four weeks after operation and total tumor growth was determined. E-cadherin, beta1 integrin, and CD44 were assessed immunohistochemically on the tumor tissue. RESULTS: The expression of E-cadherin was significantly reduced to 46.7% (complete loss of staining) in the taurolidine/heparin group. Although no significant difference was detected concerning the beta1-integrin and CD44 expression, a slightly reduced expression level with 26.7% of negative staining in metastases of the taurolidine/heparin group was observed. The total tumor weight (171.1 +/- 181.2 mg) as well as the total number of tumor lesions was also reduced by the substances compared to the control group (283.2 +/- 91.4 mg). CONCLUSIONS: Taurolidine/heparin led to a significant reduction of local tumor growth. Additionally a reduction of the expression of E-cadherin was observed. However, the biological behavior of this molecule is multivariant, controversial and still unclear. Further studies should elucidate its role in the epithelial tumor genesis. PMID- 17109892 TI - The first Susac's syndrome case in Turkey. AB - Susac's syndrome is a vasospastic disease resulting from bilateral microangiopathy of the brain, cochlea and retina. It is characterized by encephalopathy, bilateral sensorineural fluctuating hearing loss and visual loss. It is very uncommon and usually affects women during young adulthood. Since all three symptoms of the triad may not be present, the clinical diagnosis is difficult. Therefore, neuroimaging, particularly magnetic resonance imaging, has an important role in establishing the diagnosis. In this case report, we present a young woman who had all the symptoms of Susac's syndrome. This is the first Susac's syndrome case reported in Turkey. PMID- 17109893 TI - Excessive daytime sleepiness and on-road driving performance in patients with Parkinson's disease. AB - The impact of excessive daytime sleepiness (EDS) on road test performance was examined in patients with Parkinson's disease (PD). Twenty-one patients with PD completed the Epworth Sleepiness Scale (ESS) and an on-road driving test. Five participants had EDS according to their self-report on the ESS. Neither EDS nor PD medications were associated with on-road driving performance. These findings suggest that in this pilot study EDS did not impair PD patients' driving skills on a formal driving evaluation. PMID- 17109894 TI - Allelic variation investigation of the estrogen receptor within an Australian multiple sclerosis population. AB - Multiple Sclerosis (MS) is a central nervous system (CNS) chronic inflammatory demyelinating disease leading to various neurological disabilities. The disorder is more prevalent for women with a ratio of 3:2 female to male. OBJECTIVES: To investigate variation within the estrogen receptor 1 (ESR1) polymorphism gene in an Australian MS case-control population using two intragenic restriction fragment length polymorphisms; the G594A located in exon 8 detected with the BtgI restriction enzyme and T938C located in intron 1, detected with PvuII. One hundred and ten Australian MS patients were studied, with patients classified clinically as Relapsing Remitting MS (RR-MS), Secondary Progressive MS (SP-MS) or Primary Progressive MS (PP-MS). Also, 110 age, sex and ethnicity matched controls were investigated as a comparative group. No significant difference in the allelic distribution frequency was found between the case and control groups for the ESR1 PvuII (P = 0.50) and Btg1 (P = 0.45) marker. Our results do not support a role for these two ESR1 markers in multiple sclerosis susceptibility, however other markers within ESR1 should not be excluded for potential involvement in the disorder. PMID- 17109895 TI - Culture and inattentional blindness: a global workspace perspective. AB - A recent 'necessary conditions' mathematical treatment of Baars' global workspace consciousness model, analogous to Dretske's communication theory analysis of high level mental function, is used to explore the effects of embedding cultural heritage on inattentional blindness. Culture should express itself quite distinctly in this basic psychophysical phenomenon across a great variety of sensory modalities because the limited syntactic and grammatical bandpass of the rate distortion manifold characterizing conscious attention must conform to topological constraints generated by cultural context. PMID- 17109896 TI - A neutral metapopulation model of biodiversity in river networks. AB - In this paper, we develop a stochastic, discrete, structured metapopulation model to explore the dynamics and patterns of biodiversity of riparian vegetation. In the model, individual plants spread along a branched network via directional dispersal and undergo neutral ecological drift. Simulation results suggest that in comparison to 2-D landscapes with non-directional dispersal, river networks with directional dispersal have lower local (alpha) and overall (gamma) diversities, but higher between-community (beta) diversity, implying that riparian species are distributed in a more localized pattern and more vulnerable to local extinction. The relative abundance patterns also change, such that higher percentages of species are in low-abundance, or rare, classes, accompanied by concave rank-abundance curves. In contrast to existing theories, the results suggest that in river networks, increased directional dispersal reduces alpha diversity. These altered patterns and trends result from the combined effects of directionality of dispersal and river network structure, whose relative importance is in need of continuing study. In addition, riparian communities obeying neutral dynamics seem to exhibit abrupt changes where large tributaries confluence; this pattern may provide a signature to identify types of interspecific dynamics in river networks. PMID- 17109897 TI - Appetitive flight patterns of male Agrotis segetum moths over landscape scales. AB - An analysis is presented of the first harmonic radar studies of pheromone-plume locating flights of male Agrotis segetum moths over distances of up to 500 m. Upon release most moths flew in a direction having a downwind component. The first significant changes in flight orientations occur in the immediate vicinity of a pheromone source. Moths that were initially flying downwind change course and start flying crosswind whilst those that initially flew crosswind change course and start flying upwind. It is shown that such behaviour is consistent with the adoption of an effective plume-location strategy, and conditions are identified when downwind flights would be more advantageous than crosswind ones. Additionally, some of the complex flight patterns that can arise at later times are shown to be compatible with the adoption of an optimal biased scale-free (Levy-flight) searching strategy. It is found that disruptive doses of sex pheromone can have a marked influence upon male moth flight patterns. PMID- 17109898 TI - Effect of p53 haploinsufficiency on melphalan-induced genotoxic effects in mouse bone marrow and peripheral blood. AB - Mice heterozygous for a p53 null mutation develop tumours induced by genotoxic carcinogens with a shorter latency than wild type mice and have been proposed as an alternate animal model for carcinogenicity testing. Some literature data suggest that p53+/- mice might also be more sensitive to the short-term effects of genotoxic agents and manifest a haploinsufficiency phenotype that could contribute to the higher tumour susceptibility. We have compared the induction of micronuclei in bone marrow and blood of p53+/- and p53+/+ isogenic mice after treatment with a single or multiple doses of melphalan (MLP), a crosslinking genotoxic carcinogen. We have also characterized the mechanism of micronucleus induction with CREST staining of kinetochore proteins to distinguish between chromosome break- and chromosome loss-induced micronuclei. Significant increases of micronucleated bone marrow polychromatic erythrocytes and blood reticulocytes were induced under all MLP exposure conditions. The frequency of micronucleated blood erythrocytes increased linearly with duration of exposure. Micronuclei were essentially a consequence of chromosome break events. After a single MLP dose, a significant reduction of the frequency of polychromatic erythrocytes in bone marrow of p53+/+ animals suggested the induction of cytotoxicity/cell cycle delay. This effect was not observed in p53+/- mice. We believe this finding to provide some evidence of a haploinsufficiency phenotype in the modulation of cell cycle/apoptotic pathways mediated by the p53 protein. In bone marrow of wild type mice, an increased effect of multiple MLP doses was detected over that of a single administration, whereas, in p53+/- mice, no differential effect was found of different exposure durations. Possibly, the probability of micronucleus formation increased under chronic exposure because of increased cell division in response to peripheral anemia and a reduction of p53 protein level had a small effect on cell cycle modulation and on such indirect mechanism of micronucleus induction. However, pairwise comparisons between the frequencies of cells with micronuclei in wild type and p53+/- mice under all exposure conditions did not show statistically significant differences, suggesting that the observed effects of p53 haploinsufficiency were weak and temporary and a higher/faster induction of irreversible chromosome damage could not account for the increased susceptibility of p53+/- mice to MLP-induced tumours. PMID- 17109899 TI - Recovery from aphasia as a function of language therapy in an early bilingual patient demonstrated by fMRI. AB - Knowledge about the recovery of language functions in bilingual aphasic patients who suffer from left-hemispheric stroke is scarce. Here, we present the case of an early bilingual patient (German/French) with chronic aphasia. Functional magnetic resonance imaging (fMRI) was used to investigate neural correlates of language performance during an overt picture naming task in German and French (a) 32 months after stroke to assess differential recovery of both languages as a function of the preceding language therapy that was provided exclusively in German and (b) after additional short-term intensive (German) language training. At the first investigation behavioral performance confirmed selective recovery of German naming ability which was associated with increased functional brain activation compared to the French naming condition. Changes in behavioral performance and brain activation pattern as disclosed by fMRI after an additional experimental treatment were confined to the trained (German) language and indicate bilateral neuroplastic reorganization. No generalization to the untrained (French) language was observed. The present case results demonstrate use and/or training-dependent differential recovery of expressive language functions and an enhanced pattern of brain activation as a function of the rehabilitation efforts that were focussed exclusively on the patient's German language abilities. PMID- 17109900 TI - The perception of emotion and social cues in faces. PMID- 17109901 TI - Refuge use: a conflict between avoiding predation and losing mass in lizards. AB - Prey often respond to predation risk by increasing refuge use, but this strategy may entail a loss of body condition. Factors responsible for this loss of body condition remain unclear. Also how prey deal with refuge use to cope with predation risk without incurring costs of body condition, and how initial body condition affects refuge use remain barely known. We analyzed in the field whether adult Iberolacerta cyreni lizards modify their escape strategies and refuge use in areas with different levels of habitat deterioration and ecotourism pressure, which represent different levels of predation risk, and the consequences of changes in antipredator behavior to body condition. Lizards inhabiting deteriorated areas, where risk is higher, remained closer to refuges, but decreased time spent hidden in refuges after attacks, probably to maintain similar body condition than lizards inhabiting natural areas. We performed two laboratory experiments to isolate potential costs of refuge use that might affect the body condition of male lizards: a) a decrease of the efficiency of digestion due to low temperatures inside refuges and/or b) a reduction in food intake. Results suggest that refuge use is costly in terms of body condition due to reduced food intake, but the loss of efficiency of digestion seems unimportant. Lizards modified refuge use in relation to their body condition, with lizards with worse condition decreasing time hidden after predatory attacks. We concluded that lizards compensated for increased predation risk with flexible antipredatory strategies, coping with risk without incurring costs for body condition. PMID- 17109902 TI - Quaternary protoberberine alkaloids. AB - This contribution reviews some general aspects of the quaternary iminium protoberberine alkaloids. The alkaloids represent a very extensive group of secondary metabolites with diverse structures, distribution in nature, and biological effects. The quaternary protoberberine alkaloids (QPA), derived from the 5,6-dihydrodibenzo[a,g]quinolizinium system, belong to a large class of isoquinoline alkaloids. Following a general introduction, the plant sources of QPA, their biosynthesis, and procedures for their isolation are discussed. Analytical methods and spectral data are summarized with emphasis on NMR spectroscopy. The reactivity of QPA is characterized by the sensitivity of the iminium bond CN(+) to nucleophilic attack. The addition of various nucleophiles to the protoberberine skeleton is discussed. An extended discussion of the principal chemical reactivity is included since this governs interactions with biological targets. Quaternary protoberberine alkaloids and some related compounds exhibit considerable biological activities. Recently reported structural studies indicate that the QPA interact with nucleic acids predominantly as intercalators or minor groove binders. Currently, investigations in many laboratories worldwide are focused on the antibacterial and antimalarial activity, cytotoxicity, and potential genotoxicity of QPA. PMID- 17109903 TI - Arbuscular mycorrhizal symbiosis and plant aquaporin expression. AB - Almost all land plants have developed a symbiosis with arbuscular mycorrhizal fungi. Establishment of the association is accompanied by structural changes in the plant root. During arbuscule formation fungal hyphae penetrate the root apoplast and install highly specialized interfaces for solute transport between plant and fungus. The periarbuscular membrane which is part of the plant plasma membrane surrounding arbuscular structures was shown to harbour a high density of different transport systems. Among these also expression of aquaporins was described, which potentially can act as a low affinity transport system for ammonia or ammonium. The present study provides data for expression, localization and function of plant aquaporins in the periarbuscular membrane of mycorrhizal Medicago truncatula plants. PMID- 17109904 TI - Antibacterials and modulators of bacterial resistance from the immature cones of Chamaecyparis lawsoniana. AB - As part of an on-going project to characterize compounds from immature conifer cones with antibacterial or modulatory activity against multidrug-resistant (MDR) strains of Staphylococcus aureus, eight compounds were isolated from the cones of Chamaecyparis lawsoniana. The active compounds were mainly diterpenes, with minimum inhibitory concentrations ranging from 4 to 128 microg/ml against MDR effluxing S. aureus strains and two epidemic methicillin-resistant (EMRSA) clinical isolates. The compounds extracted were the diterpenes ferruginol, pisiferol and its epimer 5-epipisiferol, formosanoxide, trans-communic acid and torulosal, the sesquiterpene oplopanonyl acetate and the germacrane 4beta hydroxygermacra-1(10)-5-diene. Some of these compounds also exhibited modulatory activity in potentiating antibiotic activity against effluxing strains and ferruginol, used at a sub-inhibitory concentration, resulted in an 80-fold potentiation of oxacillin activity against strain EMRSA-15. An efflux inhibition assay using an S. aureus strain possessing the MDR NorA efflux pump resulted in 40% inhibition of ethidium bromide efflux at 10 microM ferruginol (2.86 microg/ml). We report the (1)H and (13)C NMR data for the cis A/B ring junction epimer of pisiferol which we have named 5-epipisiferol. We also unambiguously assign all (1)H and (13)C NMR resonances for trans-communic acid. PMID- 17109905 TI - Health inequalities--a sustainable development issue. PMID- 17109906 TI - Sequential decision-making in a variable environment: modeling elk movement in Yellowstone National Park as a dynamic game. AB - We develop a suite of models with varying complexity to predict elk movement behavior during the winter on the Northern Range of Yellowstone National Park (YNP). The models range from a simple representation of optimal patch choice to a dynamic game, and we show how the underlying theory in each is related by the presence or absence of state- and frequency-dependence. We compare predictions from each of the models for three variables that are of basic and applied interest: elk survival, aggregation, and use of habitat outside YNP. Our results suggest that despite low overall forage depletion in the winter, frequency dependence is crucial to the predictions for elk movement and distribution. Furthermore, frequency-dependence interacts with mass-dependence in the predicted outcome of elk decision-making. We use these results to show how models that treat single movement decisions in isolation from the seasonal sequence of decisions are insufficient to capture landscape scale behavior. PMID- 17109907 TI - The expression of Hedgehog genes (Ihh, Dhh) and Hedgehog target genes (Ptc1, Gli1, Coup-TfII) is affected by estrogenic stimuli in the uterus of immature female rats. AB - The objective of this study was to investigate the effects of estrogen receptor (ER) agonists and an ER antagonist on the expression of Hedgehog genes (Indian hedgehog: Ihh; Desert hedgehog: Dhh) and Hedgehog target genes (Patched 1: Ptc1; glioma-associated oncogene homolog 1: Gli1; chicken ovalbumin upstream promoter transcription factor II: Coup-TfII) in the rat uterus. Immature female rats were administered once with 17alpha-ethynyl estradiol (EE, an ER agonist), propyl pyrazole triole (PPT, an ERalpha-selective agonist), diarylpropionitrile (DPN, an ERbeta-selective agonist), or ICI 182,780 (an ER antagonist). Expression of mRNA for Ihh, Dhh, and Ptc1 was dose-dependently downregulated by EE in the uterus of immature rats, mediated by ER as confirmed by coadministration of ICI 182,780. The mRNA expression levels of Ptc1, Gli1, and Coup-TfII were simultaneously downregulated during the period in which the mRNA expression levels of Ihh and Dhh were downregulated in the uterus after administration of EE. PPT downregulated the transcription of Ihh, Dhh, Ptc1, Gli1, and Coup-TfII, indicating that expression of these genes was regulated by the ERalpha-dependent pathway. DPN also downregulated the transcription of Ihh and Dhh, although the effect was weaker than that of PPT, indicating that the regulation of uterine Ihh and Dhh transcription was also affected by the ERbeta-dependent pathway. These results suggest that the expression of Hedgehog genes (Ihh, Dhh) and Hedgehog target genes (Ptc1, Gli1, Coup-TfII) is affected by estrogenic stimuli in the uterus of immature female rats. PMID- 17109908 TI - TCDD decreases ATP levels and increases reactive oxygen production through changes in mitochondrial F(0)F(1)-ATP synthase and ubiquinone. AB - Mitochondria generate ATP and participate in signal transduction and cellular pathology and/or cell death. TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) decreases hepatic ATP levels and generates mitochondrial oxidative DNA damage, which is exacerbated by increasing mitochondrial glutathione redox state and by inner membrane hyperpolarization. This study identifies mitochondrial targets of TCDD that initiate and sustain reactive oxygen production and decreased ATP levels. One week after treating mice with TCDD, liver ubiquinone (Q) levels were significantly decreased, while rates of succinoxidase and Q-cytochrome c oxidoreductase activities were increased. However, the expected increase in Q reduction state following TCDD treatment did not occur; instead, Q was more oxidized. These results could be explained by an ATP synthase defect, a premise supported by the unusual finding that TCDD lowers ATP/O ratios without concomitant changes in respiratory control ratios. Such results suggest either a futile cycle in ATP synthesis, or hydrolysis of newly synthesized ATP prior to release. The TCDD-mediated decrease in Q, concomitant with an increase in respiration, increases complex 3 redox cycling. This acts in concert with glutathione to increase membrane potential and reactive oxygen production. The proposed defect in ATP synthase explains both the greater respiratory rates and the lower tissue ATP levels. PMID- 17109909 TI - Arsenic-induced bladder cancer in an animal model. AB - Dimethylarsinic acid (DMA(V)) is carcinogenic to the rat urinary bladder, but not in mice. The carcinogenic mode of action involves cytotoxicity followed by regenerative cell proliferation. Dietary DMA(V) does not produce urinary solids or significant alterations in urinary composition. The cytotoxicity is due to formation of a reactive metabolite, likely dimethylarsinous acid (DMA(III)), concentrated and excreted in the urine. Urinary concentrations of DMA(III) are dose-dependent, and the urinary concentrations are at cytotoxic levels based on in vitro studies. The no observed effect level (NOEL) in these rat dietary studies for detectable levels of DMA(III), cytotoxicity, and proliferation is 2 ppm, with marginal changes at 10 ppm. The tumorigenic dose is 100 ppm. Recent investigations have demonstrated that arsenicals administered to the rat result in binding to a specific cysteine in the hemoglobin alpha chain as DMA(III), regardless of the arsenical being administered. Monomethylarsonic acid (MMA(V)) is not carcinogenic in rats or mice. In short term experiments (< or =10 weeks), sodium arsenate in the drinking water induces significant cytotoxicity and regenerative proliferation. There is little evidence that the cytotoxicity produced following administration of arsenicals is caused by oxidative damage, as antioxidants show little inhibitory activity of the cytotoxicity of the various arsenicals either in vitro or in vivo. In summary, the mode of action for DMA(V) induced bladder carcinogenesis in the rat involves generation of a reactive metabolite (DMA(III)) leading to cytotoxicity and regenerative proliferation, is a non-linear process, and likely involves a threshold. Extrapolation to human risk needs to take this into account along with the significant differences in toxicokinetics and toxicodynamics that occur between different species. PMID- 17109910 TI - Effects of combinations of Fusarium mycotoxins on the inhibition of macromolecular synthesis, malondialdehyde levels, DNA methylation and fragmentation, and viability in Caco-2 cells. AB - We studied the interactive effects of either binary or tertiary mixtures of Fusarium mycotoxins, deoxynivalenol (DON), zearalenone (ZEA), and fumonisin B1 (FB1) on the human intestinal cell line, Caco-2, using the endpoints including malonedialdehyde (MDA) production, inhibition of protein and DNA syntheses, DNA methylation, DNA fragmentation, and cell viability as measured by the neutral red (NR) test. The mixtures of mycotoxins reduce cellular viability in increasing order: [FB1+ZEA]<[FB1+DON]<[ZEA+DON]<[FB1+DON+ZEA] in NR test. Because FB1 antagonizes the effects of estrogenic Zearalenone, FB1 was assayed against estradiol. In NR assay, mixture of FB1 and estradiol and/or ZEA improves Caco-2 cells viability in contrast to individual effects. Mixtures of ZEA or FB1 and DON, display synergistic effects in lipid peroxidation. The ability of the toxins to inhibit DNA synthesis is 45%, 70%, and 43% for 10 microM of ZEA, DON, and FBI, respectively. Their binary mixtures (at 10 microM each), inhibit DNA synthesis by 35%, 62%, and 65%, far less than additive effects. Surprisingly, the tertiary mixture (10 microM each) only inhibits DNA synthesis by 25%. ZEA, DON, and FB1 induce DNA fragmentation individually. However, mixtures of these mycotoxins always damage DNA to a greater extent. Each individual mycotoxin (10 microM) raises the percentage of 5-methylcytosine (m5dC) in DNA from 4.5% to 9%, while the combination does not increase this rate any further. Altogether, the data indicate that mixtures of Fusarium toxins are able to induce lipid peroxidation, DNA damage, DNA fragmentation, DNA methylation, and cytotoxicity in Caco-2 cells, and suggest a potential promoter effect in human intestinal cells. PMID- 17109912 TI - Soluble microbial products in membrane bioreactor operation: Behaviors, characteristics, and fouling potential. AB - This paper presents an experimental study on soluble microbial products (SMP) in membrane bioreactor (MBR) operation at different sludge retention times (SRTs). A laboratory-scale MBR was operated at SRT of 10, 20, and 40 days for treatment of readily biodegradable synthetic wastewater. The accumulation, composition, characteristics, and fouling potential of SMP at each SRT were examined. It was found that accumulation of SMP in the MBR became more pronounced at short SRTs. Carbohydrates and proteins appeared to be the components of SMP prone to accumulate in the MBR compared with aromatic compounds. The proportions of SMP with large molecular weight in supernatants and in effluents were almost identical, implying that membrane sieving did not work for most SMP. In addition, the majority of SMP was found to be composed of hydrophobic components, whose proportion in total SMP gradually increased as SRT lengthened. However, fouling potentials of SMP were relatively low at long SRTs. The hydrophilic neutrals (e.g., carbohydrates) were most likely the main foulants responsible for high fouling potentials of SMP observed at short SRTs. PMID- 17109911 TI - Characterization of HIV-1 integrase N-terminal mutant viruses. AB - During infection, human immunodeficiency virus type 1 integrase engages a number of molecules and mechanisms, both of viral and cellular origin. In one of such instances, integrase is thought to be degraded by the N-end rule proteasome pathway a process that targets the N-terminal residue of its substrates. Here we describe the properties of HIV-1 viruses in which the first amino acid residue of integrase has been substituted to render it resistant to the N-end rule pathway. As result of this exchange, we observe a set of class I and class II defects that result in a large decrease of viral replication efficiency. Specifically, reverse transcription and integration are the steps that appear to be affected. We propose that the severe deficiency of these mutants exert a strong selective pressure that leads to the near total conservation of the N-terminal residue of integrase in HIV-1, HIV-2 and SIV. PMID- 17109913 TI - Methanogenic population dynamics and performance of an anaerobic membrane bioreactor (AnMBR) treating swine manure under high shear conditions. AB - A 6-L, completely mixed anaerobic bioreactor with an external ultrafiltration membrane module was operated for 300 days to evaluate the startup and performance of an anaerobic membrane bioreactor (AnMBR) treating swine manure. The reactor had a successful startup at the initial loading rate of 1g volatile solids (VS)/L/day. After a two-fold increase in loading rate followed by a sudden, two fold increase in flow velocity through the membrane module on day 75, the performance of the AnMBR deteriorated as measured by volatile fatty acid (VFA) accumulation, decrease in pH, and decrease in biogas production. The methanogenic population dynamics in the reactor were monitored with terminal restriction fragment length polymorphism (T-RFLP). Changes in the relative levels of Methanosarcinaceae and Methanosaetaceae were consistent with changes in VFA concentrations, i.e., high and low levels of acetate corresponded to a high abundance of Methanosarcinaceae and Methanosaetaceae, respectively. The levels of hydrogenotrophic methanogens of the order of Methanomicrobiales increased during decreased reactor performance suggesting that syntrophic interactions involving hydrogenotrophic methanogens remained intact regardless of the degree of shear in the AnMBR. PMID- 17109914 TI - Ammonia removal from pretreated methane fermentation effluent through a soil trench system: a column experiment. AB - In order to find the optimal running conditions and mechanisms of ammonia removal through a soil trench system that is designed for treating pretreated methane fermentation effluent, a soil column whose structure was similar to the soil trench system was prepared, and irrigated with wastewater below 30 degrees C. At the beginning, ammonia was mainly adsorbed by the soils, and the ammonia adsorption capacity of soils gradually saturated. After the 12th day, nitrification began in the soil column; the ammonia in the soil column decreased sharply, and the nitrite and nitrate peak appeared sequentially as the wastewater application rate decreased from 0.74 to 0.37 l h(-1). When the nitrification in the soil column reached a steady-state, 98% of all the ammonia in the influent was transformed into nitrate. By changing the running conditions such as temperature, aeration, and wastewater application rate, it was found that the ammonia removal efficiency can be improved by aeration and impeded by low temperature. In these three variables, wastewater application rate has much greater affect on the ammonia removal efficiency; a lower wastewater application rate can increase the ammonia removal efficiency substantially because of the longer travel time. PMID- 17109915 TI - Thermal degradation of PCDD/F, PCB and HCB in municipal solid waste ash. AB - A thermal degradation procedure for reducing the concentrations of mono- to octa chlorinated PCDD/Fs, PCBs and hexa-chlorobenzenes (HCB) in filter ash from incinerated municipal solid waste (MSW) is described. Thermal treatment of filter ash samples at 500 degrees C for 60 min in a closed system providing low oxygen conditions resulted in 97% and 99% reductions in the total and I-TEQ concentrations of PCDD/Fs, to 6.8 microg kg(-1) ash and <0.05 microg I-TEQ kg(-1) ash, respectively. Increasing the thermal treatment time to 480 min, at the same temperature, yielded 99% reductions in both total and I-TEQ concentrations of the mono- to octa-chlorinated PCDD/Fs. Similar effects were observed for HCB and PCBs. The data from this study indicate that PCDD/Fs and other toxic organic compounds in ash from incinerated MSW, can be effectively degraded by this procedure, which combines relatively low-temperatures, short treatment times, and low oxygen conditions. PMID- 17109916 TI - Low cost sorbents for the removal of methyl parathion pesticide from aqueous solutions. AB - Sorptive potential of selected agricultural waste materials i.e. rice (Oryza sativa) bran (RB), bagasse fly ash (BFA) of sugarcane (Saccharum officinarum), Moringa oleifera pods (MOP) and rice husk (RH) for the removal of methyl parathion pesticide (MP) from surface and ground waters has been investigated. Optimization of operating parameters of sorption process, i.e. sorbent dose, agitation time, pH, initial concentration of sorbate, and temperature have been studied. The sorption data fitted to Freundlich, Langmuir and Dubinin Radushkevich (D-R) sorption isotherms. The maximum capacities of RB, BFA, MOP and RH for MP were calculated to be 3.6+/-0.8, 5.3+/-1.4, 5.2+/-1.5 and 4.7+/-1.0 mmolg(-1) by Freundlich, 0.39+/-0.009, 0.39+/-0.005, 0.36+/-0.004 and 0.35+/ 0.008 mmolg(-1) by Langmuir and 0.9+/-0.08, 1.0+/-0.10, 1.0+/-0.10 and 0.9+/-0.07 mmolg(-1) by D-R isotherms respectively, employing 0.1g of each sorbent, at pH 6, 90 min agitation time and at 303 K. Application of first order Lagergren and Morris-Weber equations to the kinetic data yielded correlation coefficients, close to unity. Thermodynamic parameters of sorption process, i.e. DeltaH, DeltaS and DeltaG were computed and their negative values indicated the exothermic and spontaneous nature of sorption process. The pesticide may be stripped by sonication with methanol, making the regeneration and reutilization of sorbents promising. The sorbents investigated exhibited their potential applications in water decontamination, treatment of industrial and agricultural waste waters. PMID- 17109917 TI - Changes of polycyclic aromatic hydrocarbons during composting of sewage sludges with chosen physico-chemical properties and PAHs content. AB - The aim of the experiment was to determine changes in the PAH content during sewage sludge composting. The extent was evaluated to which sewage sludge properties influences on PAHs losses during composting. Five municipal sewage sludges were studied from sewage treatment plants located in the south-east of Poland, i.e. in Stalowa Wola, Krasnik, Lublin, Zamosc and Bilgoraj. The treatment plants selected were characterised by their differentiation with catchments area as well as the industrial character of the areas. Sludge composting was carried out in aerobic conditions in plastic containers for a period of 76 days. The content of 14 polycyclic aromatic hydrocarbons (PAHs) and some physical-chemical (TOC, Nt, TOC/Nt, CEC, BS, TEB, pH, available potassium and phosphorus, total forms of cations) sewage sludge properties were monitored. In the present study correlations between PAHs losses (during various phases of composting) and physical-chemical properties of sewage sludges and composts were also evaluated. The sewage sludges were characterised both by differentiated properties and by PAH content. The total PAH content (the sum of 14 PAHs from the EPA List with exception of naphthalene and acenaphthene) in the sludges ranged from 3674 to 11236 microg kg(-1). Four- and three-ring compounds were predominant in the sewage sludges. Almost all of the sludges were characterised by differentiated content levels of both individual PAHs groups and PAHs sum. After 76 days of composting, a significant lowering of the PAH content was observed in all sludges as compared to the beginning of the experiment. The range of total observed PAH losses was from 15.8% to 48.6%, depending on the sewage sludge. The largest lowering of the PAH content was observed in the sludge with the lowest concentration of the xenobiotics studied. The largest losses in individual PAHs were observed in the case of five- and six-rings compounds, and were probably related to strong pollutant binding by other components of the sewage sludge. The few significant relationships showed that PAH losses during composting were only slightly determined by the analysed physical-chemical properties. PMID- 17109918 TI - Pollutants in Hong Kong soils: polycyclic aromatic hydrocarbons. AB - An extensive soil survey was carried out to study the polycyclic aromatic hydrocarbon (PAH) contaminations in 138 soil samples collected throughout Hong Kong. Results demonstrated that there were low levels of PAH contaminations (median of summation operator 16US EPA PAHs=140 microg kg(-1)) for all land uses (urban park, greening area, country park, rural area, restored landfill, agricultural farmland, orchard farm, crematorium, industrial and near highway area). However, localized hotspots were identified with summation operator 16PAH concentrations as high as 19,500 microg kg(-1) in one urban park. These findings were also confirmed by multivariate analysis. Comparison of PAH profiles showed a widespread domination of its 4-ring member. The major contribution was vehicular emissions from petroleum, and however at the hotspots, the improper disposal of used motor oils. In general, the pollution levels for all the land uses were below the recommended values for residential and general purposes stated in soil quality guidelines such as Netherlands and Denmark except certain identified hotspots. The potential health hazards imposed by these hotspots were alarming, and their existence (3 out of 138 samples) suggested that sole monitoring of atmospheric PAHs may not adequately address the hidden risks to human in urban city. PMID- 17109919 TI - Polybrominated diphenyl ethers in blood from Korean incinerator workers and general population. AB - This study was conducted to examine PBDE exposure in Koreans, with a special focus on incinerator workers due to their potential for occupational exposure to PBDEs. A total of 92 blood samples from 30 incinerator workers, 51 nearby residents and 11 controls were analyzed. The mean total PBDE concentration calculated from the 13 most concentrated congeners for all samples was 16.84+/ 7.48 ng/g lipid, which was somewhat higher concentration than in other countries except North America and Canada. The PBDE levels and congener profiles detected in incinerator workers were not distinctly different from those found in the general population. In all groups tested, BDE-47 was dominant (mean contribution=32.5%) followed by BDE-153 (23.6%) and relatively high portions of BDE-183 (16.5%) were found. No strong trend was observed between PBDE levels and a number of key biological factors examined in this study, however, weak correlations were observed in PBDE levels measured against dietary habits, particularly in fish consumption frequency and gender. Overall, our data suggest that the occupational exposure of incinerator workers to PBDEs can be considered minor, while other lifestyle factors can have a greater contribution to PBDE exposure. PMID- 17109920 TI - Cadmium availability to wheat grain in soils treated with sewage sludge or metal salts. AB - Grain Cd concentrations were determined in wheat (Triticum aestivum L.) grown in 1999, 2001 and 2003, at six sludge cake field experiments. Three of these sites also had comparisons with Cd availability from metal amended liquid sludge and metal salts. Grain Cd concentrations in all years and at all sites were significantly linearly correlated with NH4NO3 extractable Cd and soil total Cd (P<0.001). Soil extractability was greater in the liquid sludge and metal salt experiments than in the cake experiments, as were grain Cd concentrations. Across all the sites, NH4NO3 extractable soil Cd was no better at predicting grain Cd than soil total Cd. Stepwise multiple linear regression analysis showed that soil total Cd, pH and organic carbon were the only significant (P<0.001) variables influencing wheat grain Cd concentrations, explaining 78% of the variance across all field experiments (1408 plots). This regression predicted that the current UK soil total Cd limit of 3 mg kg(-1) was not sufficiently protective against producing grain above the European Union (EU) grain Cd Maximum Permissible Concentration (MPC) of 0.235 mg Cd kg(-1) dry weight, unless the soil pH was > 6.8. Our predictions show that grain would be below the MPC with > 95% confidence with the proposed new EU draft regulations permitting maximum total Cd concentrations in soils receiving sludge of 0.5 mg kg(-1) for soils of pH 5-6, 1 mg kg(-1) for soils of pH 6-7, and 1.5 mg kg(-1) for soils of pH > or = 7. PMID- 17109921 TI - Vertical profile of PCDD/Fs, dioxin-like PCBs, other PCBs, PAHs, chlorobenzenes, DDX, HCHs, organotin compounds and chlorinated ethers in dated sediment/soil cores from flood-plains of the river Elbe, Germany. AB - Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) and other organic micropollutants were determined in dated sediment/soil cores collected from the flood-plain of the river Elbe near Pevestorf (PT), approximately 125 km upstream of Hamburg, and Heuckenlock (HL) in southeast of Hamburg. Concentrations of PCDD/Fs peaked sharply at PT in the 1950s and at HL at the end of the 1940s. Cluster analyses provide evidence that the region of Bitterfeld-Wolfen (about 350-400 km upstream of Hamburg) could be the source of the PCDD/F contamination existing in the cores PT and HL since the 1940s. Obviously it is caused by sediments of the river Elbe of a similar composition. Whereas the PCDD/Fs, HCHs (hexacyclohexane isomers), DDX (DDT, DDD, DDE), and tetrachlorinated ethers in PT and HL presumably originated predominantly from the Bitterfeld-Wolfen region, organotin compounds in HL and dichlorinated haloethers in HL during the 1940s and 1950s can probably largely be attributed to emissions from the Hamburg region. Although they are separated by a large distance, in both sediment cores PT and HL concentrations and composition patterns of most organic micropollutants analyzed widely match. Inductively it can be concluded that similar contaminations will be found in many of the river bank soils between the Bitterfeld-Wolfen region and Hamburg. Excavation of top soils may uncover highly contaminated materials. Since the dated sediment cores show the variation in contaminants in the Elbe sediments over a defined time period, it is possible to make an approximate assessment of the actual degree of contamination to be expected in areas where in previous decades contaminated dredged sediments from the Elbe and from the Port of Hamburg have been deposited on land and used for building plots or for agricultural purposes. PMID- 17109922 TI - Evaluation of the leachability of polychlorinated dibenzo-p-dioxins and dibenzofurans in raw and solidified air pollution control residues from municipal waste incinerators. AB - Leachability of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) from raw and solidified air pollution control (APC) residues with selected solvents, including acetic acid, simulated acid rain, humic acid, linear alkylbenzene sulfonate (LAS) and n-hexane was investigated. High-chlorinated PCDD/F congeners were observed in all leachates of raw APC residue samples, with the largest total leaching concentration (61.60 ngm(-3); 0.30 ngI-TEQm(-3)) from treatment with humic acid. Low-chlorinated congeners were mainly leached with LAS and n-hexane. Solidification and stabilization (S/S) processes with cement and sulfur-containing chelating agent decreased the leachability of PCDD/Fs by up to 98% with humic acid and LAS as solvents. However, S/S processes enhanced the leachability of both high- and low-chlorinated PCDD/F congeners with n-hexane as the solvent, which largely increased the toxic equivalent quantity of leachates. These results suggest that conventional S/S processes may effectively restrain the release of PCDD/Fs when APC residues are leached with rain water or natural organic compounds (e.g., humic acid), but may have a deteriorated effect when APC residues are leached with nonpolar organic solvents (e.g., n-hexane) coexisting in the landfill sites. PMID- 17109923 TI - Removal of PAHs from water using an immature coal (leonardite). AB - It has been studied an immature coal (leonardite) as an adsorbent for removing PAHs [fluorene, pyrene, benzo(k)fluoranthene, benzo(a)pyrene and benzo(g,h,i)perylene] from water. To determine the efficiency of leonardite as an adsorbent of PAHs, factors such as pH, contact time and equilibrium sorption were evaluated in a series of batch experiments. There were no significant differences in the removal percentages for the various pH values studied, except for fluorene. The adsorption of fluorene was higher at lower pH values. The equilibrium time was reached at 24h. At this time, more than 82% of the pyrene, benzo(k)fluoranthene, benzo(a)pyrene and benzo(g,h,i)perylene had been removed. During the first 2h, the adsorption rate increased rapidly. After that time, however, there was a minor decrease. Equilibrium data were fitted to Freundlich models to determine the water-leonardite partitioning coefficient. Physical adsorption caused by the aromatic nature of the compounds was the main mechanism that governed the removal process. The polarity of the humic substances in leonardite may also have influenced the adsorption capacity. PMID- 17109924 TI - The influence of radical architecture on cadmium bioaccumulation in the black mangrove, Avicennia germinans L. AB - Two groups of Avicennia germinans plants with differences in the radical architecture were exposed under hydroponic conditions to 95ppm of cadmium (Cd) for a period of 24h. Later, Cd concentration in roots, stems and leaves was determined by graphite furnace atomic absorption spectrophotometry. Our results showed that, for both groups of plants, the roots accumulated higher concentration of Cd as compared to stems and leaves, though, the plants of group B displayed enhanced radical architecture, better growth performance, and lower Cd concentration as compared to plants of group A. In contrast, low values of leaves/roots Cd transportation index, and bioaccumulation factor were found in plants of group B. These results suggest that the higher radical architecture developed in plants of group B might better adjust the uptake of Cd as a result of an integrated network of multiple response processes for instances, production of organic acids, antioxidative replay, cell-wall lignification and/or suberization. Further studies will be focused in understanding the role of the radical system in mangrove plants with the rhizosphere activation and root adsorption to soil Cd under natural conditions. PMID- 17109925 TI - Vertical distributions of persistent organic pollutants (POPs) caused from organochlorine pesticides in a sediment core taken from Ariake bay, Japan. AB - Persistent organic pollutants (POPs) of organochlorine pesticides such as dichlorodiphenyl trichloroethane (DDTs) and its metabolites, hexachlorobenzene (HCB), heptachlor, chlordane compounds (CHLs), aldrin, dieldrin, endrin, mirex and isomers of hexachlorocyclohexane (HCHs), were analyzed in sediment cores collected from Ariake Bay, Japan. Although Chikugo River has the largest flow in Ariake Bay, the total concentration of POPs in surface sediment was found in St.3, mouth of the Yabe River, where that level was two times higher than St.1, mouth of the Chikugo River. It could be assumed that the potential source of POPs contamination is relatively close to this study area. Relatively high residue levels of HCB, HCHs and DDTs in sediment core were found during the period from 1967 to 1970. In this layer, the HCB concentration investigated in sediment cores of Ariake Bay was maximum, 2.6 ng g(-1) dry weight and higher than that of Tokyo Bay and subsequently detected in deeper sediment cores. The ratio of (DDD+DDE)/SigmaDDTs was high in top sediments (0-2 cm). Although there was a concentration of DDTs in the top sediment, it was noticed that DDT emission did not cause direct input of DDT recently. The HCHs determined in upper sediment cores might be originated from long-range accumulation in environment after dispersing of technical-grade HCH. Moreover, the CHLs, dieldrin and heptachlor concentrations were detected and recently increased in sediment core. POPs in sediment cores of Ariake Bay based on the possibility of PCP contamination during the early 1960s were reflected. PMID- 17109926 TI - Linear QSAR regression models for the prediction of bioconcentration factors by physicochemical properties and structural theoretical molecular descriptors. AB - The development of QSAR models useful for the prediction of fish bioconcentration factor (BCF) for a wide range of different chemical classes is crucial for the assessment and prioritisation of potentially persistent bioaccumulative and toxic substances. In this study we present QSAR models for BCF developed on a wide range of chemical structural classes of environmental and toxicological interest (such as dyes and various chlorinated and brominated compounds). The aim is to provide valid QSAR models, statistically validated for predictivity, for the prediction of BCF in general, but also for problematical chemical classes such as highly hydrophobic chemicals. Several descriptors, calculated by different commercially available software packages, have been employed in order to take into account relevant information provided by physicochemical properties (octanol/water partition coefficient and water solubility) and molecular features (structural and quantum-chemical molecular descriptors). The best descriptor subsets for the models were selected using the Genetic Algorithm-Variable Subset Selection strategy (GA-VSS) and calculations were performed by ordinary least squares regression. Starting from a data set of 640 compounds (logK(ow) range from -2.34 to 12.66), we developed linear QSARs, firstly for a data set of 620 compounds (logK(ow) range from -2.34 to 10.35) and secondly specifically for 87 highly hydrophobic chemicals (logK(ow) range from 6.00 to 10.35). All these models have been statistically validated (both internally by cross-validation and bootstrap and externally, by "a priori" splitting of available data by Kohonen Map-ANN in training and prediction sets) and their structural chemical domain has been verified by the leverage approach. PMID- 17109927 TI - Changes in photochemical and antioxidant enzyme activities in maize (Zea mays L.) leaves exposed to excess copper. AB - Changes in photosynthetic and antioxidant activities in maize (Zea mays L.) leaves of cultivars 3223 and 31G98 exposed to excess copper (Cu) were investigated. Cu treatment reduced the shoot and root length of both cultivars. No significant difference of Cu accumulation in the roots of both cultivars was observed while the cultivar 3223 accumulated significantly higher Cu in leaves than 31G98. The observed decreases in effective quantum efficiency of PSII, ETR and qP indicate an over excitation of photochemical system in 3223 compared to 31G98. The leaf chlorophyll and carotenoid contents of both cultivars decreased with increasing Cu concentration. A far higher production of anthocyanins in 31G98 has been observed than that of 3223. At 1.5 mM Cu concentration, all antioxidant enzyme activities increased in leaves of the cultivar 31G98 while there were no significant changes in SOD and GR activities in 3223 compared to the control except increased APX and POD activities. The lower Cu accumulation in leaves and higher antioxidant enzyme activities in 31G98 suggested an enhanced tolerance capacity of this cultivar to protect the plant from oxidative damage. PMID- 17109928 TI - Degradation of explosives-related compounds using nickel catalysts. AB - We report the ability of nickel-based catalysts to degrade explosives compounds in aqueous solution. Several nickel catalysts completely degraded the explosives, although rates varied. Nearly all of the organic explosive compounds tested, including 2,4,6-trinitrotoluene (TNT), hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), were rapidly degraded to below detection limits by a powdered nickel on an alumina-silicate support (Aldrich nickel catalyst). Perchlorate degradation was minimal (<25%). Degradation of TNT by Aldrich nickel catalyst resulted in apparent first-order kinetics. Significant gaseous 14C was released and collected in an alkaline solution (most likely carbon dioxide) from [14C]RDX and [14C]HMX, indicating heterocyclic ring cleavage. Significant gaseous 14C was not produced from [14C]TNT, but spectrophotometric evidence indicated loss of aromaticity. Degradation occurred in low ionic strength solutions, groundwater, and from pH 3 to pH 9. Degradation of TNT, RDX, and HMX was maintained in flow-through columns of Aldrich nickel catalyst mixed with sand down to a hydraulic retention time of 4h. These data indicate that nickel-based catalysts may be an effective means for remediation of energetics-contaminated groundwater. PMID- 17109929 TI - Factors affecting the degradation of amoxicillin in composting toilet. AB - The biological and non-biological factors that affect the degradation of amoxicillin in the composting process of feces have been investigated. The effect of living bacteria and the enzyme (beta-lactamase) on amoxicillin decay was examined, and our results indicated that the biological effects are likely to be negligible. Consequently, the effect of phosphate, ammonia and pH level as non biological factors was investigated by monitoring the reduction rate of amoxicillin in phosphate and ammonia buffer solutions with several pH levels. Each reduction rate constant was integrated by a simulation model, and the each calculated amoxicillin reduction profile was compared to the reduction profiles of amoxicillin in the composting process of feces. The calculated results corresponded almost exactly to the experimental profiles. We therefore concluded that the degradation of amoxicillin in a toilet matrix was dependent on the concentration of ammonia, phosphate and hydroxyl ion. PMID- 17109930 TI - Photocatalytic activity of the calcined H-titanate nanowires for photocatalytic oxidation of acetone in air. AB - Hydrogen titanate (H-titanate) nanowires were prepared via a hydrothermal reaction of TiO2 powders (P25) in KOH solutions and then calcined at various temperatures. The phase structure, crystallite size, morphology, specific surface area, and pore structures of the calcined H-titanate nanowires at various temperatures were characterized with field emission scanning electron microscope, X-ray diffraction, transmission electron microscopy and nitrogen adsorption desorption isotherms, and their photocatalytic activities were evaluated by photocatalytic oxidation of acetone in air. With increasing calcination temperature, the specific surface area and porosity of the calcined samples steadily decreased. At a calcination temperature range of 400-600 degrees C, the calcined H-titanate nanowires showed higher photocatalytic activity than P25 powders for photocatalytic oxidation of acetone. Especially, at 500 degrees C, the calcined H-titanate nanowires showed the highest photocatalytic activity, which exceeded that of P25 by a factor of about 1.8 times. This can be attributed to the synergetic effect of larger specific surface area, higher pore volume and the presence of brookite TiO2. With further increase in the calcination temperature (700-900 degrees C), the photocatalytic activity of the samples decreased obviously owing to the growth of TiO2 crystallites. PMID- 17109931 TI - Modification of the degradative capacity of a soil artificially contaminated with diesel. AB - Samples of an Ah horizon from a Cambisol under oakwood vegetation were artificially contaminated with diesel (at doses of 20, 40, 80, 160 and 400 microl g(-1)) under laboratory conditions. The presence of the contaminant caused a decrease in the microbial biomass and in phosphomonoesterase, beta-glucosidase and particularly, urease activities. In contrast, the basal respiration and mineralization of nitrogen, specifically ammonification, both increased. The microorganisms that survived in the contaminated soil showed increased metabolic activity, as a result of their use of dead microorganisms as a substrate and of a lack of competition. Although the results indicate that the properties related to the degradative capacity of the soils varied differently in response to the contamination, the use of a biochemical quality index (including different biochemical properties), which was designed for estimating the degree of deterioration of the soil, revealed a clear decrease in the biochemical quality of the diesel-contaminated soils. PMID- 17109932 TI - Assigning concentration values for dioxin and furan congeners in human serum when measurements are below limits of detection: an observational approach. AB - Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in dietary sources for humans have been declining over the previous two decades. These declines have been accompanied by decreases in concentrations of these compounds in humans, as evidenced by measurements in blood and milk. Because of the decreasing concentrations of PCDD/PCDFs in the environment and in humans, measuring PCDD/PCDF congeners in humans has become increasingly difficult, despite advances in analytic methods. An observational approach was recently described to address the quandary of non-detectable results in determining toxic equivalents. This approach, called the congener ratio approach, is specifically for cases where concentrations of 2,3,7,8-TCDD (TCDD) are below the limit of detection (LOD), and where concentrations of 1,2,3,7,8 pentachlorodibenzo-p-dioxin (PeCDD) are equal to or above the LOD. Development of this approach relied on evaluating data on measured concentrations of TCDD and PeCDD in human serum from the general population. The congener ratio approach for TCDD and PeCDD was based on the concentration of TCDD being approximately 40% that of PeCDD in serum from the general population. Additional analyses presented here reveal that when concentrations of both congeners are above the LOD, the data appear to generally support the congener ratio approach for TCDD and PeCDD, with the caveat that gender may affect the ratio. However, the TCDD/PeCDD relation is less clear when TCDD is less than the LOD; in this situation, the relation overpredicts levels of TCDD approximately 80% of the time for the 2001 2002 NHANES database. Using the congener ratio approach for other PCDD/PCDF congeners requires assessing the correlation and the frequency of detection for both TCDD and PeCDD. PMID- 17109933 TI - PAHs in the bulk atmospheric deposition of the Seine river basin: source identification and apportionment by ratios, multivariate statistical techniques and scanning electron microscopy. AB - The origin of polycyclic aromatic hydrocarbons (PAH) contamination in bulk atmospheric deposition at two sites of the Seine estuary, one urban and one industrial, has been investigated. The PAH profiles indicate that PAHs mainly have a pyrolytic origin, both in urban and industrial areas. PAH sources vary during the year with an increase of high molecular weight PAH proportions (especially for carcinogenic PAHs) in winter, that means an increase of combustion processes such as domestic heating. Ratios of indicator PAHs (FTH/FTH+PYR and IcdP/IcdP+BghiP) confirm the pyrolytic origin of PAHs. In summer, ratios show the presence of industrial sources. In addition to these two methods, a factor analysis/multiple linear regression model was applied and gave an approximation of PAH source apportionment. PAH were found to be associated predominantly with emissions from road traffic (gasoline and diesel), that accounts for 17-34%. Domestic heating is a very important PAH source in urban areas and accounts for up to 85% of PAHs in winter. Industrial emissions (refineries...) account for 25% in the industrial area in summer. Each is an identified source category for the region and these results are consistent with fly-ashes identified by scanning electron microscopy. This study demonstrates that a combination of source identification methods is a far more efficient than one method alone. PMID- 17109934 TI - Bioavailability and plant accumulation of heavy metals and phosphorus in agricultural soils amended by long-term application of sewage sludge. AB - Amendment of agricultural soils with municipal sewage sludges provides a valuable source of plant nutrients and organic matter. Nevertheless, addition of heavy metals and risks of eutrophication continue to be of concern. Metal behaviour in soils and plant uptake are dependent on the nature of the metal, sludge/soil physico-chemical properties and plant species. A pot experiment was carried out to evaluate plant production and heavy metal uptake, soil heavy metal pools and bioavailability, and soil P pools and possible leaching losses, in agricultural soils amended with sewage sludge for at least 10 years (F20) compared to non amended soils (control). Sewage sludge application increased soil pH, N, Olsen extractable-P, DOC and exchangeable Ca, Mg and K concentrations. Total and EDTA extractable soil concentrations of Cu and Zn were also significantly greater in F20, and soil metal (Cu, Mn and Zn) and P fractionation altered. Compared to the control, in F20 relative amounts of acid-extractable (Mn, Zn), reducible (Mn, Zn) and oxidisable (Cu, Zn) metal fractions were greater, and a dominance of inorganic P forms was observed. Analyses of F20 soil solutions highlighted risks of PO4 and Cu leaching. However, despite the observed increases in metal bioavailability sewage sludge applications did not lead to an increase in plant shoot concentrations (in wild plants or crop species). On the contrary, depending on the plant species, Mn and Zn tissue concentrations were within the deficiency level for most plants. PMID- 17109935 TI - Ghrelin stimulates growth hormone secretion and food intake in aged rats. AB - Age-related decreases in energy expenditure have been associated with the loss of skeletal muscle and decline of food intake, possibly through a mechanism involving changes of growth hormone (GH) secretion and feeding behavior. Age related declines of growth hormone secretion and food intake have been termed the somatopause and anorexia of ageing, respectively. Ghrelin, a 28-amino-acid peptide, was isolated from human and rat stomachs as an endogenous ligand of growth hormone secretagogue receptor. Ghrelin stimulates growth hormone release and food intake when peripherally administered to rodents and humans. Here, we investigate the relationship between age-related decline of growth hormone secretion and/or food intake and ghrelin function. Ghrelin (10 nmol/kg body weight) was administered intravenously to male 3-, 12-, 24-and 27-month-old Long Evans rats, after which growth hormone concentrations and 2 h food intake were measured. An intravenous administration of ghrelin to rats increased food intake in all generations. In addition, to orexigenic effect by ghrelin, intravenous administration of ghrelin elicited a marked increase in plasma GH levels, with the peak occurring 15 min after administration. These findings suggest that the aged rats maintain the reactivity to administered exogenous ghrelin. PMID- 17109936 TI - Clopidogrel resistance? AB - Clopidogrel is an effective inhibitor of platelet activation and aggregation due to its selective and irreversible blockade of the P2Y(12) receptor. Combination antiplatelet therapy with clopidogrel and aspirin is an important strategy for patients with acute coronary syndromes and those undergoing percutaneous interventions. Despite significant benefits demonstrated with combination antiplatelet treatment in large clinical trials, the occurrence of adverse ischemic events, including stent thrombosis, remains a serious clinical problem. Recent studies have demonstrated distinct response variability and nonresponsiveness to clopidogrel therapy based on ex vivo platelet function measurements. Small scale investigations have suggested that nonresponsiveness may be associated with a heightened risk for adverse clinical events. The above findings have stimulated a close examination of clopidogrel metabolism. PMID- 17109937 TI - Melatonin protects against epirubicin-induced cardiotoxicity. AB - We investigated the cytoprotective effect of melatonin in epirubicin-induced cardiotoxicity using four experimental groups of male Wistar rats: untreated control rats, epirubicin-treated rats, epirubicin+melatonin-treated rats, and melatonin-treated rats. We examined the histopathological and biochemical effects of melatonin on the epirubicin-induced changes and measured the levels of the lipid peroxidation end-product (malondialdehyde, MDA), an indicator of nitric oxide (NO) synthesis (nitrite/nitrate production), and reduced glutathione (GSH) in the heart. We also studied the extracellular matrix components (fibronectin, laminin) in the heart. Vacuole formation, mitochondrial deformation and degeneration, and disordered myofibrillary structures were detected ultrastructurally in the epirubicin-treated group. The degeneration was reduced in the heart tissues of the epirubicin+melatonin group. Epirubicin increased the nitrite/nitrate production, but did not change the MDA and GSH levels significantly. Melatonin treatment lowered the nitrite/nitrate concentrations, while increasing the GSH levels, which exceeded the levels in epirubicin+melatonin-treated rats. We conclude that the epirubicin increased the nitrozative stress, not the oxidative stress, in heart tissue, and the cardioprotective effect of melatonin was partially attributed to the suppression of epirubicin-induced nitrozative stress. These results suggest that melatonin partially protects against epirubicin-induced cardiotoxicity. PMID- 17109938 TI - Body mass index: relationship to clinical, pathologic and features of microsatellite instability in endometrial cancer. AB - OBJECTIVES: There is a well known association between obesity and endometrial cancer. We sought to examine the relationships between body mass index (BMI), as a measure of obesity, and known demographic, clinical, and molecular characteristics of microsatellite instability and MLH1 promoter methylation in a cohort of patients with endometrial cancer. METHODS: Corpus cancer specimens were prospectively obtained from 473 consecutively enrolled patients between 1992 and 2004. Clinical and pathologic data were extracted from review of the medical record. Microsatellite instability (MSI) was evaluated in all tumors, and methylation of the MLH1 promoter was determined for MSI positive tumors. RESULTS: The median (SD) age and BMI were 64.8 years (11.9) and 33.5 (9.4), respectively. Histology included 376 endometrioid (79%), 69 serous/clear cell or mixed (15%), and 28 sarcomas (6%). Median BMI was 32.4 for endometrioid, 31.0 for serous/clear cell or mixed, and 27.8 for sarcomas (p=0.14). BMI was negatively associated with age at surgery (p<0.01). The remainder of analyses excluded sarcoma histology. BMI was associated with stage of disease; patients with stage I/II disease had significantly higher BMI than those with stage III/IV disease (32.6 vs. 30.6; p=0.02). In relation to molecular features of endometrial cancer, BMI was significantly different between MSI positive tumors compared to MSI negative tumors (30.3 vs. 32.7; p=0.02). MSI was also significantly different between tumor histology, occurring with a higher frequency in Type I than Type II tumors (p<0.01). CONCLUSIONS: The majority of endometrial cancer patients are obese. Those with higher BMI are more likely to be younger, present with early stage disease, and have MSI negative tumors. PMID- 17109939 TI - Regulation of 5-HT2A/C receptors and DOI-induced behaviors by protein kinase Cgamma. AB - Protein kinase Cgamma (PKCgamma) null mutant mice demonstrate increased behavioral impulsivity and ethanol consumption. Pharmacological studies have shown that 5-HT(2A/C) receptors modulate impulsivity and ethanol consumption in rodents and that PKC can regulate 5-HT(2A/C) receptors. To determine whether PKCgamma plays a selective role in 5-HT(2A/C) receptor regulation, biochemical and behavioral experiments were performed in PKCgamma mutant and wild-type mice. DOI-stimulated phosphoinositol hydrolysis and [(125)I]-DOI saturation binding in the PFC, and quantitative autoradiography of [(125)I]-DOI binding sites in 15 brain regions were analyzed. DOI-induced head twitch responses (HTR) were measured in naive mice after an acute 2.5 mg/kg injection of DOI. Results indicated that DOI-induced HTR was significantly greater in mutant mice compared to wild-type mice. Results of the phosphoinositol hydrolysis, membrane binding, and autoradiography experiments indicated that in mutant mice, increased HTR was associated with increased 5-HT(2A/C) receptor function in the PFC, but not increased receptor number or affinity suggesting that PKCgamma regulates receptor function but not receptor number. These data support a role for 5-HT(2A/C) receptors in the PFC in mediating some of the behavioral differences observed between PKCgamma mutant and wild-type mice. PMID- 17109940 TI - Early deprivation increases exploration and locomotion in adult male Wistar offspring. AB - The aim of the study was to investigate the effects of repeated early maternal deprivation (individual separation in warm or cold environment for 4 h/day) during postnatal days 1-15 on emotional responses in novel situations and voluntary alcohol consumption in adult male Wistar rat offspring. Brain monoamine levels and plasma levels of corticosterone were measured at the end of the experiment. Controls were exposed to a brief (3 min) daily handling procedure. As adults, both groups of early deprived rats showed increased nose poking and locomotion in the exploration test compared to controls. Moreover, separated rats kept in room temperature also showed increased locomotion when tested for an extended period of time. There were no differences in alcohol intake, monoamine levels, or corticosterone levels between early deprived animals and controls. In addition, the dams' retrieval behavior of pups was studied, showing that dams of early deprived pups spent more time in the nest with the pups after the 4-h separation period compared to control dams. Our results indicate that early deprived animals show decreased emotionality in novel settings compared to briefly handled controls. Furthermore, the present study demonstrates that methodological issues within the maternal separation paradigm may be influential factors for behavioral changes in adulthood. PMID- 17109941 TI - Corticotropin-releasing factor receptor type 1, but not type 2, in the ventromedial hypothalamus modulates dopamine release in female rats. AB - Corticotropin-releasing factor (CRF) plays an important role in stress responses and is mediated through two subtypes of receptors, CRF receptor type 1 (CRFR1) and CRF receptor type 2 (CRFR2). Each CRF receptor might have a different function through several neurotransmitter systems; however, the mechanism remains unclear. To clarify the role of each receptor in dopamine (DA) metabolism, we measured the change of extracellular concentrations of DA and the metabolites in the ventromedial hypothalamus (VMH) that played important roles in the stress response of freely moving female rats in response to the direct administration of comparative CRFR1 selective agonist, CRF, or CRFR2 selective agonist, Urocortin II (Ucn II), into the brain region. Administration of 10 microg CRF increased extracellular concentrations of DA compared with 2 microg CRF immediately after injection, and this effect was not observed after 60 min of 10 microg CRF injection. On the other hand, this change did not always occur after Ucn II administration. These results suggest that the activation of CRFR1, but not CRFR2, modulates the release of DA in VMH. PMID- 17109942 TI - The effects of histaminergic agents in the dorsal hippocampus of rats in the elevated plus-maze test of anxiety. AB - High levels of histamine are found in the hippocampus. The central histamine system is involved in many physiological behavioural processes including anxiety related behaviours both in animals and humans. In the present study, we investigated the effects of intra-hippocampal CA1 (intra-CA1) microinjection of histaminergic agents on anxiety-related behaviours in rats, using elevated plus maze test of anxiety. Intra-CA1 administration of histamine (at the dose of 10 microg/rat) increased open arm time (%OAT) and open arm entry (%OAE) but not locomotor activity, thus showing an anxiolytic response. Intra-CA1 microinjection of pyrilamine (H1 receptor antagonist; at the doses of 10, 20 and 40 microg/rat) in combination with histamine (10 microg/rat) showed a decrease in the %OAT and %OAE. Higher dose of the antagonist (40 microg/rat) by itself increased both %OAT and %OAE, but not locomotor activity, indicating an anxiolytic effect. Intra-CA1 microinjection of ranitidine (H2 receptor antagonist), at the doses of 10, 20 and 40 microg/rat, also reduced the histamine response. Furthermore, the H2 receptor antagonist by itself reduced %OAT and %OAE without affecting locomotor activity. The results may indicate an anxiogenic effect for the antagonist. Our results showed that histamine may modulate anxiety via H1 and H2 receptors in the CA1 region of hippocampus of the rat. PMID- 17109943 TI - The role of delta opioid receptors in the anxiolytic actions of benzodiazepines. AB - The anxiolytic effects of benzodiazepines appear to involve opioid processes in the amygdala. In previous experiments, overexpression of enkephalin in the amygdala enhanced the anxiolytic actions of the benzodiazepine agonist diazepam in the elevated plus maze. The effects of systemically administered diazepam are also blocked by injections of naltrexone into the central nucleus of the amygdala. The current studies investigated the role of delta opioid receptors in the anxiety-related effects of diazepam. Three days following bilateral stereotaxic injections of viral vectors containing cDNA encoding proenkephalin or beta-galactosidase (control vector), the delta opioid receptor antagonist naltrindole (10 mg/kg, s.c.) attenuated the enhanced anxiolytic effects of 1-2 mg/kg diazepam in rats overexpressing preproenkephalin in the amygdala. Despite this effect, naltrindole failed to attenuate the anxiolytic action of higher diazepam doses (3 mg/kg) in animals with normal amygdalar enkephalin expression. Similarly, the mu opioid receptor antagonist, beta-funaltrexamine (20 mg/kg, s.c.), had no effect on the anxiolytic effect of diazepam alone. These data support a role for delta opioid receptors in the opioid-enhanced anxiolytic effects of diazepam. PMID- 17109944 TI - The protective effect of frontal cortex dehydroepiandrosterone in anxiety and depressive models in mice. AB - We aimed to verify whether DHEA, a neuoroactive neurosteroid, has a protective role in preventing the occurrence or enhancement of the severity of depression and anxiety in mice. Four groups were tested: controls, mice possessing significantly high frontal cortex DHEA levels, achieved by repeated DHEA injections (1.6 mg/Kg, i.p.), mice that have significantly low frontal cortex DHEA levels, consequent to castration and mice possessing significantly low frontal cortex DHEA levels, treated with DHEA to reverse its level to normal, achieved by castration and repeated DHEA injections (0.4 mg/Kg, i.p.). The Forced Swim Test to determine depressive-like and the Elevated Plus Maze (EPM) to evaluate anxiety-like behaviors, were used. We found that DHEA had an anti depressive-like effect, as shown by a decreased immobility time in mice possessing a high level of frontal cortex DHEA and increased immobility time in mice that have a low frontal cortex DHEA level. DHEA also demonstrated an anti anxiety-like effect, as shown by the open-arm time in EPM, which correlated with DHEA level. Mice with significantly low DHEA levels when restored to normal, did not differ from controls. In conclusion, high levels of DHEA have an anti-anxiety like and an anti-depressive-like effect in mice and those with low levels of frontal cortex DHEA are more vulnerable to depression and/or anxiety. PMID- 17109945 TI - Comparison of meat composition from offspring of cloned and conventionally produced boars. AB - This study compares the meat composition of the offspring from boars produced by somatic cell nuclear transfer (n=4) to that of the offspring from conventionally produced boars (n=3). In total, 89 commercial gilts were artificially inseminated and 61 progressed to term and farrowed. All of the resulting piglets were housed and raised identically under standard commercial settings and slaughtered upon reaching market weight. Loin samples were taken from each slaughtered animal and shipped offsite for meat composition analysis. In total, loin samples from 404 animals (242 from offspring of clones and 162 from controls) were analyzed for 58 different parameters generating 14,036 and 9396 data points from offspring of clones and the controls, respectively. Values for controls were used to establish a range for each parameter. Ten percent was then added to the maximum and subtracted from the minimum of the control range, and all results within this range were considered clinically irrelevant. Of the 14,036 data points from the offspring of clones, only three points were found outside the clinically irrelevant range, two of which were within the range established by the USDA National Nutrient Database for Standard Reference, Release 18, 2005; website: (www.nal.usda.gov/fnic/foodcomp/search/). The only outlier was the presence of Eicosadienoic acid (C20:2) in one sample which is typically present in minute quantities in pork; no reference data were found regarding this fatty acid in the USDA National Nutrient Database. In conclusion, these data indicated that meat from the offspring of clones was not chemically different than meat from controls and therefore supported the case for the safety of meat from the offspring of clones. PMID- 17109946 TI - Quantification of embryo quality by respirometry. AB - It is generally accepted that assessment of embryo metabolism, in particular oxygen consumption, may improve embryo selection by identifying the embryos with higher developmental competence. Several methods have been employed to measure embryonic oxygen consumption, but most of them were detrimental to subsequent embryo development. Recently, we have introduced the Nanorespirometer system, which is a non-invasive and highly sensitive technology developed for the individual measurement of embryonic respiration rates. This technology is able to perform single measurements at a fixed time or stage of embryonic development without adversely influencing embryo viability. Concomitantly, and based on the same principles, a second technology -- the Embryo Respirometer -- has been developed. The Embryo Respirometer allows the continuous measurement of individual respiration rates with simultaneous acquisition of digital images of each embryo, during the entire culture period (6-7 days). In this review, both technologies are described and their potential use as diagnostic tools for improving embryo selection in bovine and human following IVF treatments is discussed. Correlations between respiration rates of individual embryos and other parameters such as morphological quality, sex, stage of development, kinetics, diameter, expression of key metabolic genes and subsequent viability following embryo transfer are also examined. On the basis of the results obtained, it is postulated that assessment of embryonic respiration rates in association with other viability parameters allows for a more accurate embryo evaluation, both under clinical and research conditions. PMID- 17109947 TI - The effect of type of vaginal insert and dose of pLH on embryo production, following fixed-time AI in a progestin-based superstimulatory protocol in Nelore cattle. AB - The objective was to analyze and report field data focusing on the effect of type of progesterone-releasing vaginal insert and dose of pLH on embryo production, following a superstimulatory protocol involving fixed-time artificial insemination (FTAI) in Nelore cattle (Bos taurus indicus). Donor heifers and cows (n = 68; 136 superstimulations over 2 years) received an intravaginal, progesterone-releasing insert (CIDR or DIB, with 1.9 or 1.0 g progesterone, respectively) and 3-4 mg of estradiol benzoate (EB) i.m. at random stages of the estrous cycle. Five days later (designated Day 0), cattle were superstimulated with a total of 120-200 mg of pFSH (Folltropin-V), given twice daily in decreasing doses from Days 0 to 3. All cattle received two luteolytic doses of PGF2alpha at 08:00 and 20:00 h on Day 2 and progesterone inserts were removed at 20:00 h on Day 3 (36 h after the first PGF2alpha injection). Ovulation was induced with pLH (Lutropin-V, 12.5 or 25 mg, i.m.) at 08:00 h on Day 4 with FTAI 12, 24 and in several cases, 36 h later. Embryos were recovered on Days 11 or 12, graded and transferred to synchronous recipients. Overall, the mean (+/-S.E.M.) number of total ova/embryos (13.3 +/- 0.8) and viable embryos (9.4 +/- 0.6) and pregnancy rate (43.5%; 528/1213) did not differ among groups, but embryo viability rate (overall, 70.8%) was higher in donors with a DIB (72.3%) than a CIDR (68.3%, P = 0.007). In conclusion, the administration of pLH 12 h after progesterone removal in a progestin-based superstimulatory protocol facilitated fixed-time AI in Nelore donors, with embryo production, embryo viability and pregnancy rates after embryo transfer, comparable to published results where estrus detection and AI was done. Results suggested a possible alternative, which would eliminate the need for estrus detection in donors. PMID- 17109948 TI - Reversible reproductive control in harbour seals (Phoca vitulina) with a gonadotropin-releasing hormone agonist. AB - Reproductive control in captive pinnipeds is an important management subject for many facilities. To date reproductive control in harbour seals (Phoca vitulina) has been achieved using anti-androgens, progestagen preparations, castration, and physical separation of the sexes. The harbour seal group at the seal station in Friedrichskoog, Germany consists of three mature females (all >10 years), one older mature male (13 years of age in 2000) and one male who reached maturity during the study (3 years of age in 2000). In 2000 the older mature male received for the first time a 3-month depot injection of a gonadotropin-releasing hormone agonist (buserelin acetetate, 9.9 mg) by subcutaneous injection. This male was subsequently given the gonadotropin-releasing hormone agonist in 2001, 2004 and 2005. The younger male reached maturity during the investigation and received burserelin for the first time in 2004 and again in 2005. No pups were born in 2001, 2002, 2005 or 2006. No reproductive control was performed in 2002 and 2003, resulting in three newborns in 2003 and 2004. Serum levels of testosterone were measured by a routine liquid chromatography coupled with mass spectometry. Pre burserelin testosterone levels varied between 0.02 and 2.18 ng/ml. Post burserelin levels were under the detection limit except for the first year of the investigation. No behavioural changes such as changes in social ranking and no clinical side effects were observed. This study shows that the gonadotropin releasing hormone agonist, burserelin acetate, can be used for reversible reproductive control in harbour seals without observed side effects or detrimental behavioural changes. PMID- 17109949 TI - Pregnancy percentage following deposition of sex-sorted sperm at different sites within the uterus in estrus-synchronized heifers. AB - Our objective was to assess the effect on heifer pregnancy rate of deposition at three sites within the uterus of frozen-thawed sex-sorted sperm at a fixed time after estrus synchronization. Estrus was synchronized in 209 heifers by administration of PGF2a 14 days apart. At 80-82 h after the second PGF2a injection, X-chromosomes bearing fractions of semen with 2.2 x 10(6) sperm in insemination dose were used for single insemination into the uterine body (UB-AI, n=91) or for intracornual deposition in the middle of the uterine horn (MH-AI, n=57) or close to the utero-tubal junction (UTJ-AI, n=61). The overall pregnancy rate was 43.1%. Pregnancy rates did not differ (P>0.05) among sites of sperm sperm deposition, between the two farms at which the heifers were kept or between the two bulls producing the semen. Within UB-AI, MH-AI and UTJ-AI treatments, pregnancy rates were 41.8%, 49.1% and 39.3%, respectively (P>0.05). Pooled across classes for deposition site, pregnancy rate was 25.1% higher (P<0.01) for heifers showing strong signs of estrus than for heifers showing weak signs of estrus (45.9 versus 20.8%, respectively). Embryonic and fetal loss from diagnosis of pregnancy to term and at calving equalled 5.6%. Of 88 calves of identified sex, 93.2% were female. In conclusion, pregnancy rates of heifers did not differ significantly following deposition of 2.2 x 10(6) sex-sorted sperm 80-82 h after the second PGF2a injection near the utero-tubal junction, in the middle of the horn or into the uterine body. PMID- 17109950 TI - Surface microhardness of enamel after different home bleaching procedures. AB - OBJECTIVES: The purpose of this study was to evaluate the influence of different home bleaching procedures on surface microhardness of human enamel. METHODS: Among eight groups 192 incisors were distributed. The facial surface of each incisor was polished and baseline hardness of enamel (m0; Knoop) was assessed with a load of 1N for 30s. Subsequently, the enamel was treated for 14 days with the bleaching agent: groups 1, 2 and 4 Viva Style Paint on, 8% carbamide peroxide (CP) 1x20min, 2x20min and 2x5min; group 3 Colgate Simply White, 5.9% hydrogen peroxide (HP), 2x30min; group 5 Viva Style 10% CP 1x1h; group 6 Blend-a-med White Strips, 5.9% HP 2x30min; group 7 Odol-med3 Beauty-Kur, sodium chlorite 2x10min; group 8 control, running water 1x1h. Hardness was reassessed after the last bleaching treatment (m1) and after 6 weeks storage in artificial saliva (m2). RESULTS: Changes in microhardness were as follows (m0-m1): (1) -2.3 (+/-20.3); (2) -8.9 (+/-27.2); (3) 63.4 (+/-56.3); (4) 9.6 (+/-30.1); (5) 12.8 (+/-62.6); (6) 92.2 (+/-50.2); (7) 158.4 (+/-59.7); (8) 10.6 (+/-38.5). Statistical analysis showed that hardness values were significantly (p< or =0.0005; Wilcoxon test) reduced in groups 3, 6, and 7 (m1) and in group 7 (m2). SIGNIFICANCE: Both type of bleaching agent and concentration have a significant influence on the microhardness of enamel. The most critical bleaching agent seems to be the one containing sodium chlorite in combination with citric acid. PMID- 17109951 TI - Polymerization kinetics of dental adhesives cured with LED: correlation between extent of conversion and permeability. AB - OBJECTIVES: The aim of this study was to analyze the polymerization kinetics of different adhesive films in relation to their permeability after exposure to different LED curing units. METHODS: One adhesive from each class was analyzed: a three-step etch-and-rinse (OptiBond FL; Sybron-Kerr), a two-step etch-and-rinse (One-Step, Bisco), a two-step self-etch (Clearfil Protect Bond, Kuraray) and a one-step self-etch adhesive (Xeno III; Dentsply DeTrey). Adhesive films were prepared and cured with SmartLite IQ (Dentsply) or L.E. Demetron I (Demetron Kerr) up to complete curing. Polymerization kinetic curves of the tested adhesives were obtained with differential scanning calorimetry (DSC). In particular, total reaction time and extent of polymerization (Ep) at 20, 40 or 60s were compared. Permeability of the adhesive films was evaluated on flat dentin surfaces of human extracted teeth connected to a permeability device and statistically analyzed. RESULTS: Total reaction time differed among the adhesives tested: OptiBond FLor=50% of samples; p,p'-DDE was the most frequent (96.03%), followed by endosulphan-diol (76.86%) and lindane (74.17%). Presence of more pesticides was significantly associated with lower birth weight. Mean concentration of p,p'-DDE was 2.37+/ 2.80 ng/g of placenta or 76.62+/-104.85 ng/g of lipid. Higher maternal body mass index was significantly associated with higher endosulphan concentrations in placenta, and greater maternal weight gain was significantly associated with higher p,p'-DDE concentrations. Prenatal exposure to OC xenoestrogens may be a causative factor in adverse reproductive health trends, and further studies are required to identify and describe pathways of this exposure to enhance preventive measures. PMID- 17109957 TI - The fundamental contribution of Jan C. Waldenstrom to the discovery and study of the so-called "Waldenstrom macroglobulinemia". AB - Waldenstrom macroglobulinemia may be said to be characterized cheifly by two features: (1) a striking increase in IgM globulin of the "monoclonal gammapathy" type and (2) an abnormal bone marrow characterized by either a leukemic or leukemic-like picture, in which lymphocytes of various types are prominent. All the other features, hemorrhagic, hemolytic, rheologic (viscosity syndrome) must be considered as secondary manifestations. This historical review article summarizes the fundamental contribution of Jan Costa Waldenstrom to the discovery and study of this lymphoproliferative disorder. PMID- 17109958 TI - Learning from tragedy: a survey of child and adolescent restraint fatalities. AB - OBJECTIVE: This descriptive study examines 45 child and adolescent fatalities related to restraints in residential (institutional) placements in the United States from 1993 to 2003. METHOD: The study team used common Internet search engines as its primary case discovery strategy to determine the frequency and the nature of the fatalities, as well as the characteristics of the children and the adolescents involved. RESULTS: Male children and adolescents were over represented in the study sample. Thirty-eight of the fatalities occurred during or after a physical restraint, and 7 fatalities occurred during the use of mechanical restraints. Twenty-eight of the deaths occurred in a prone restraint. In 25 of the fatalities, asphyxia was the cause of death. CONCLUSION: In the 23 cases in this study where information is available, none of the child behaviors or conditions that prompted the restraint would meet the standard of danger to self or others: the commonly accepted criteria for the use of a restraint. The study points to deficiencies in fatality reporting, recommends reporting fatalities to established state child fatality review boards, and reinforces that restraints be governed by strict protocol and monitoring. The study also urges caution to policymakers in substituting or changing restraint procedures based on the incomplete data reported in this study. PMID- 17109959 TI - Cross-cultural reliability and validity of the Multidimensional Neglectful Behavior Scale Adult Recall Short Form. AB - OBJECTIVE: To provide data on the cross-cultural reliability of the Adult Recall Short Form of the Multidimensional Neglectful Behavior Scale (MNBS), and also illustrative validity evidence. The MNBS includes items on four aspects of neglectful behavior: cognitive, emotional, physical, and supervisory. METHOD: The data were obtained by questionnaires completed by a convenience sample of 7179 students at 33 universities in 17 countries. RESULTS: The alpha coefficient of internal consistency for the entire sample was .72. By site, the coefficients ranged from one site with .55-12 sites in the .60-.69 range, and 20 sites with alpha of .70 or higher. The validity of the scale was indicated by low confounding with social desirability response set in all 33 cultural contexts, and by the results of construct validly analyses which controlled for physical maltreatment found that students who were victims of neglectful behavior as a child were more likely than other students to have physically assaulted a dating partner in the previous 12 months. CONCLUSIONS: Although the results are from a relatively privileged sample, they revealed high rates of neglectful behavior and are sufficiently promising to encourage use of the Adult Recall Short Form of the MNBS in a variety of cultural settings. PMID- 17109960 TI - Double-stranded RNA and antiviral immunity in marine shrimp: inducible host mechanisms and evidence for the evolution of viral counter-responses. AB - Double-stranded RNA (dsRNA) is a common virus-associated molecular pattern and a potent inducer of antiviral responses in many organisms. While it is clear that the specific RNA interference (RNAi) response, a phenomenon triggered by dsRNA, serves antiviral functions in invertebrates, innate (non-specific) antiviral immune reactions induced by dsRNA (e.g. the Interferon response) have long been thought to be restricted to vertebrates. Recent work in an underappreciated experimental model, the penaeid shrimp, is challenging these traditional distinctions, by demonstrating the existence of both innate (non sequence specific) and RNAi-related (sequence-specific) antiviral phenomena in crustacea. Here we discuss the evidence for this bivalent role of dsRNA in the initiation of antiviral responses in shrimp, and present new data that suggest that the antiviral functions of the shrimp RNAi machinery have imposed selective pressures on an evolving viral pathogen. These findings open the door for the discovery of novel mechanisms of innate immunity, and provide a basis for the future development of strategies to control viral diseases in the commercially important penaeid shrimp. PMID- 17109961 TI - Intra-specific variability of virulence in Leishmania infantum zymodeme MON-1 strains. AB - This study aims to characterize the intra-specific variability of virulence in Leishmania infantum zymodeme MON-1 strains isolated from dogs and immunocompetent and immunosuppressed patients through the evaluation of growth pattern, infective ability and immunopathogenicity. Two of the strains, classified as the most virulent, presented higher levels of macrophage infection, increased promastigote replication in culture medium and as well as amastigote multiplication within macrophages. These strains caused the most pathogenic infection inducing splenomegalia and maximum parasite loads in spleen and liver of BALB/c mice. The other strains exhibited either low virulence, with reduced infective capability and low replication levels, or an intermediate virulent phenotype showing mixed features similar to low and high virulent phenotypes. A correlation between the infectivity, growth dynamics and pathogenicity of each strain and the humoral and cellular immune response was demonstrated. Strains with accentuated virulent phenotype induced higher levels of anti-Leishmania IgG1 antibodies and TGF-beta but reduced production of IFN-gamma. Virulence phenotype seems to be a characteristic of each strain regardless of the host (dog or human) from which it was firstly isolated. PMID- 17109962 TI - Are there sex differences associated with the effects of ecstasy/3,4 methylenedioxymethamphetamine (MDMA)? AB - Sex has been identified as an important factor in moderating the effects of several drugs of abuse. Given the increasing popularity of ecstasy (3,4 methylenedioxymethamphetamine [MDMA]) use, it is important for researchers and clinicians to understand the factors that may influence its pharmacological actions to improve education, harm reduction and treatment efforts. This review focuses on preclinical and clinical research that examines the role of sex as an independent variable in the effects of ecstasy/MDMA. A systematic search of PsycINFO and MEDLINE electronic databases from 1966 to April 2006 was conducted. Both preclinical and clinical studies show a sexually dimorphic pattern in the acute, subacute and possibly long-term effects of ecstasy/MDMA. Specifically, adult females are more sensitive than males to the acute and subacute physical and psychological effects of ecstasy/MDMA and long-term alterations in aspects of 5-HT functioning. Conversely, males are more sensitive to the acute physiological effects of ecstasy/MDMA. These findings are consistent with research outcomes reported for other substances such as amphetamines and cocaine. Potential reasons for these sex differences and directions for future research are discussed. PMID- 17109963 TI - Candidate inhibitors of porcine complement. AB - Therapeutic complement inhibition is a promising strategy for treatment of a number of diseases as judged from rodent studies. The species distance from rodents to humans may limit the clinical relevance of these studies. The pig is an alternative animal for studies of human diseases like sepsis and ischemia/reperfusion injury. However, available complement inhibitors for use in pigs are scarce. The aim of the present study was to investigate and compare the efficacy of selected candidate inhibitors of porcine complement in vitro for possible future application in vivo. Sera from three different pigs were each incubated with three different activators of the complement system (zymosan, heat aggregated immunoglobulin G (HAIGG) and Escherichia coli). Three groups of complement inhibitor candidates were tested: serine protease inhibitors (FUT-175 and C1-inhibitor), monoclonal antibodies (anti-factor B (fB) and anti-factor D (fD)) and a recombinant regulatory protein (vaccinia virus complement control protein (VCP)). Read-out was the terminal C5b-9 complement complex (TCC). The serine protease inhibitors FUT-175 and C1-inhibitor dose-dependently inhibited TCC formation in zymosan-, HAIGG- and E. coli-activated porcine sera, but with different efficacy. Complete inhibition of TCC was obtained using 0.2 mg/mL FUT 175, but required 16 mg/mL of C1-inhibitor. The monoclonal anti-fB and -fD antibodies both inhibited TCC formation dose-dependently, but in different ways. Anti-fB at high dose (1 mg/mL) completely inhibited TCC formation in sera activated with zymosan and virtually completely in sera activated with HAIGG, but not in sera activated with E. coli. Anti-fD inhibited all three activators at low dose (0.05 mg/mL), and approximately 50% TCC reduction was obtained. The recombinant complement regulatory protein VCP efficiently and dose-dependently inhibited TCC formation with a complete inhibition found at 0.05 mg/mL for all three activators. All candidates tested inhibited porcine complement activation, but in different ways and to different degrees. Of the complement-specific candidates, VCP inhibited all activators completely at low doses. PMID- 17109964 TI - Differential disruption of nuclear volume and neuronal phenotype in the preoptic area by neonatal exposure to genistein and bisphenol-A. AB - Changes in the volumes of sexually dimorphic brain nuclei are often used as a biomarker for developmental disruption by endocrine-active compounds (EACs). However, these gross, morphological analyses do not reliably predict disruption of cell phenotype or neuronal function. In the present experiments, we used a more comprehensive approach to assess whether postnatal exposure to the EACs genistein (GEN) or bisphenol-A (BIS) affected the development of two sexually dimorphic brain regions in male rats: the anteroventral periventricular nucleus of the hypothalamus (AVPV) and the sexually dimorphic nucleus of the preoptic area (SDN). In addition to nuclear volumes, we also measured the number of immunopositive calbindin neurons in the SDN and the activational patterns of gonadotropin-releasing hormone (GnRH) neurons, a neuronal population that is functionally linked to the AVPV. In rats, exposure of the neonatal male brain to endogenous estrogen, aromatized from testicular testosterone, is essential for the proper sexual differentiation of these endpoints. Thus, we hypothesized that exposure to BIS and GEN during this critical period could disrupt brain sexual differentiation. Animals were given four subcutaneous injections of sesame oil (control), 250 microg GEN, or 250 microg BIS at 12 h intervals over postnatal days (PND) 1 and 2, gonadectomized on PND 85, and treated sequentially with estrogen and progesterone to stimulate Fos expression in GnRH neurons, a marker for their activation. A cohort of age-matched ovariectomized (OVX) females that were given the same hormone treatment in adulthood served as a positive control group. SDN volume was unchanged by treatment, but the number of calbindin neurons in the SDN was significantly increased by both BIS and GEN. GEN, but not BIS, demasculinized male AVPV volume, but patterns of GnRH neuronal activation were not affected by either compound. These results suggest that acute exposure to EACs during a critical developmental period can independently alter nuclear volumes of sexually dimorphic nuclei and their phenotypic profiles in a region specific manner. PMID- 17109965 TI - Modulation of creatine kinase activity by ruthenium complexes. AB - Creatine kinase is a crucial enzyme for brain, heart and skeletal muscle energy homeostasis, and a decrease of its activity has been associated with cell death. Many biological properties have been attributed to ruthenium complexes. In this context, this work was performed in order to evaluate creatine kinase activity from rat brain, heart and skeletal muscle (quadriceps) after administration of ruthenium complexes, trans-[RuCl(2)(nic)(4)] (nic=3-pyridinecarboxylic acid) 180.7 micromol/kg (complex I), trans-[RuCl(2)(i-nic)(4)] (i-nic=4 pyridinecarboxylic acid) 13.6 micromol/kg (complex II), trans-[RuCl(2)(dinic)(4)] (dinic=3,5-pyridinedicarboxylic acid) 180.7 micromol/kg (complex III) and trans [RuCl(2)(i-dinic)(4)] (i-dinic=3,4-pyridinedicarboxylic acid) 180.7 micromol/kg (complex IV). Our results showed that complex I caused inhibition of creatine kinase activity in hippocampus, striatum, cerebral cortex, heart and skeletal muscle. Besides, complex II did not affect the enzyme activity. complexes III and IV increased creatine kinase activity in hippocampus, striatum, cerebral cortex and heart, but not in skeletal muscle. Besides, none of the complexes in vitro altered creatine kinase activity, suggesting that enzymatic activity is indirectly affected by complexes I, III and IV. It is believed that diminution of creatine kinase in brain of rats caused by complex I may be related to results from other study reporting memory impairment caused by the same complex. Further research is necessary in order to elucidate the effects of ruthenium complexes in other important metabolic enzymes. PMID- 17109966 TI - Immune dysfunction in HIV-seronegative, Cryptococcus gattii meningitis. AB - The pathophysiology of meningitis caused by Cryptococcus gattii in apparently immunocompetent individuals remains unclear. We measured multiple cytokines in CSF from a HIV-seronegative, apparently immunocompetent, Thai patient with C. gattii meningitis, over the first 2 weeks of antifungal therapy. Levels of proinflammatory IFN-gamma, TNF-alpha, and IL-6 were very low compared to patients with HIV-related Cryptococcus neoformans meningitis and of IL-10 very high. While patients with C. gattii meningitis may be a heterogeneous group, these data suggest in this case a maladapted immune response to cryptococcal exposure had allowed progression to clinical cryptococcal disease. PMID- 17109967 TI - Auditory evoked responses in the rat: transverse mastoid needle electrodes register before cochlear nucleus and do not reflect later inferior colliculus activity. AB - A previously described technique putatively differentiates short-latency auditory evoked potentials in peripheral and central neural pathways of the mouse and rat [Galbraith G, Waschek J, Armstrong B, Edmond J, Lopez I, Liu W, et al. Murine auditory brainstem evoked response: putative two-channel differentiation of peripheral and central neural pathways. J Neurosci Methods 2006;153:214-20]. This technique involves recording from orthogonally oriented subdermal needle electrode pairs, using fast sample rates (100k/s) to accurately measure differences in neural timing and waveform morphology. Electrodes oriented in a transverse plane (mastoid-to-mastoid) register an initial positive-going peak earlier than peaks recorded from electrodes oriented along the scalp midline (anterior and posterior to the interaural line). The absolute latency of the early mastoid component is consistent with an origin in the primary auditory nerve, while delayed midline latencies implicate activity in central neural pathways. We report here the results of recording simultaneously from transverse mastoid (M) needle electrodes and electrodes acutely implanted in cochlear nucleus (CN) and inferior colliculus (IC). The results show a highly consistent pattern in which the initial mastoid component leads CN by an average of 0.16 ms, suggesting an obligatory neural site of origin of the mastoid response that is distal to IC, namely the auditory nerve. Moreover, later IC components (beyond approximately 3.5 ms) are completely absent in mastoid recordings, indicating that the transverse mastoid recordings provide a relatively isolated measure of early auditory neural activity. PMID- 17109968 TI - A survey on anxious and depressive complaints of Brazilian women. AB - BACKGROUND: Affective disorders and symptoms are very prominent among women. However specific complaints and symptoms may vary in different cultures. METHODS: A survey on the most common complaints from Brazilian women, identified as anxious or depressive symptoms, was performed with a focus group of 6 psychiatrists. The results are shown and a narrative review of manifestations of anxious and depressive symptoms in Latin American women are reported. RESULTS: Complaints and symptoms of depression and anxiety are associated with some ethnic and socio-cultural factors. The overlap between depression and anxiety is substantial. LIMITATIONS: The authors concluded that there is still insufficient quantitative scientific evidence, allowing a clear view of this subject. CONCLUSIONS: There have been suggestions that ethnic, cultural, and economic conditions seem to play an important role in presentation of depressive and anxious symptoms. PMID- 17109969 TI - Intimate partner violence and sexual coercion among pregnant women in India: relationship with depression and post-traumatic stress disorder. AB - BACKGROUND: Intimate partner violence (IPV) is prevalent in most parts of the world. It is also prevalent during pregnancy. METHODS: This study assessed the prevalence of IPV during pregnancy and evaluated its relationship with mental health outcomes, including depression and post-traumatic stress disorder (PTSD). Pregnant women (n=203) attending an antenatal clinic in a public hospital in Bangalore were assessed for presence of IPV as well as depressive, somatic, PTSD symptoms and life satisfaction. RESULTS: Self-reported physical violence in the last year was reported by 14% of women, psychological abuse by 15%, and sexual coercion by 9%. One-half of these women reported ongoing abuse during pregnancy. Depression, somatic, and PTSD symptoms were higher in those with a history of abuse or sexual coercion, and life satisfaction was poorer in those with any form of violence. Among those reporting a history of sexual coercion, severity of violence was related to increased psychiatric morbidity. Alcohol abuse in the spouse was a predictor of the presence and severity of abuse. LIMITATIONS: The study was conducted in a single clinic in southern India which is a large country with very diversified populations. CONCLUSION: The experience of intimate partner violence and its mental health consequences are quite prevalent in India which is a culture where gender disparities are normative and pregnancy is a particularly vulnerable period. PMID- 17109970 TI - Social anhedonia and schizotypy: the contribution of individual differences in affective traits, stress, and coping. AB - While social anhedonia is a promising indicator of vulnerability to schizophrenia, it remains uncertain whether anhedonia is a core feature of schizotypy or merely a secondary associated characteristic. This issue was examined by comparing dimensional scores on schizophrenia spectrum personality disorder symptoms derived from clinical interviews among three groups: a) "pure" social anhedonics with high scores on the Revised Social Anhedonia Scale (SAS; [Eckblad, M.L., Chapman, L.J., Chapman, J.P., Mishlove, M., 1982. The Revised Social Anhedonia Scale. Unpublished test, University of Wisconsin, Madison.]) and low scores on the Magical Ideation Scale (MIS, [Eckblad, M.L., Chapman, L.J., 1983. Magical ideation as an indicator of schizotypy. Journal of Consulting and Clinical Psychology, 51, 215-225.]), b) subjects with high MIS and low SAS scores, and c) controls with low scores on both scales. This study also sought to identify individual differences in stress reactivity, personality, coping style, and social support that might be related to severity of clinical symptoms among at-risk subjects. Compared to controls, the SAS group had higher levels of schizotypal, schizoid, and paranoid symptoms and the MIS group had higher schizotypal symptoms. Among social anhedonics, individual differences in perceived stress, trait negative affectivity, and coping style accounted for over 40% of the variance in schizotypal and paranoid symptoms. This cross-sectional study bolsters support for the validity of social anhedonia as a primary feature of schizotypy. Longitudinal studies are required to determine whether these individual differences potentiate clinical outcomes among social anhedonics. PMID- 17109971 TI - Heterophile antibody interference in a solid phase sandwich immunoassay for detection of equine growth hormone in plasma. AB - Heterophile antibodies (HAs) present in serum recognize animal immunoglobulins and are one of the most unpredictable causes of false results in immunoassays. However, no study has yet reported their interference on the diagnostic reliability of immunochemical analyses on horse plasma. Recently, we developed a sandwich ELISA for detection of equine growth hormone (eGH) in plasma. In a pilot study to measure basal eGH levels (blood samples were drawn from 13 horses every 10 min for 1h), we noted one horse with abnormally high eGH (>100 ng/mL). We demonstrate here that this plasma eGH level was falsely elevated due to interference from HAs. The interfering antibodies were polyspecific immunoglobulins, with fairly broad species-specificity, which affected the eGH immunoassay by bridging the mouse IgG capture antibody and the rabbit IgG conjugate. This produced artificial sandwiches which led to overestimation of the eGH plasma concentration. Spiking horse plasma with pure mouse and rabbit immunoglobulins or whole plasma of several species significantly reduced but did not totally eliminate the HAs interference. Immunoglobulins and whole plasma differed in their ability to block the interference, suggesting that HAs may recognize other proteins beside immunoglobulins in animal sera. To investigate whether HAs have any implications in equine clinical practice, we decided to seek information on the incidence of HAs interference in normal animals. We collected single plasma samples from another 114 horses and we found that 5 of these had plasma HAs. Therefore, in total 6 out of the 127 horses examined (4.7%) had plasma HAs generating falsely elevated eGH measures. In conclusion, this study provides the first evidence of HAs in horse plasma interfering with an immunoassay and indicates that veterinary surgeons and diagnostic laboratory staff should be aware of this potential for interference in tests on horse plasma using monoclonal or polyclonal antibody reagents. PMID- 17109972 TI - Production and characterization of two serotype independent monoclonal antibodies against foot-and-mouth disease virus. AB - Two foot-and-mouth disease virus (FMDV) monoclonal antibodies (mAbs) were produced from mice immunized with either FMDV serotype A, subunit (12S) or FMDV serotype O, whole virus (140S). Both mAbs (F1412SA and F21140SO) recognized all seven serotypes of FMDV in a double antibody sandwich (DAS) ELISA, suggesting that the binding epitopes of the two mAbs are conserved between serotypes. These mAbs are IgG1 isotype and contain kappa light chains. In order to define the mAb binding epitopes, the reactivity of these mAbs against trypsin-treated and denatured FMDV were examined using an indirect ELISA. The binding site of the mAb, F1412SA is trypsin sensitive and the epitope is linear. Both ELISA and Western blot results suggested that the polypeptide VP2 contributed to the immunodominant site. This mAb showed reactivity to VP2 peptide (DKKTEETTILEDRIL). The mAb, F21140SO, recognized an epitope which is trypsin resistant and discontinuous. This mAb binding to FMDV is dependent on conformational structures of intact viral (140S) or subunit (12S) particle, since it failed to recognize any viral protein in Western blot. This conformational and highly conserved epitope is the first identified epitope among all seven FMDV serotypes. Because the use of mAbs increases the specificity, accuracy and efficiency of diagnostic tests compared to polyclonal antisera, these two mAbs with different specificities are suitable for type-independent diagnosis of FMDV, such as DAS ELISA, or could be adapted to immuno-chromatographic or flow-through rapid test. PMID- 17109973 TI - Differential contributions of B7-1 and B7-2 to the development of murine experimental allergic conjunctivitis. AB - B7-1 and B7-2 are the co-stimulatory molecules that are involved in activation of T cells. We investigated whether B7-1 and B7-2 play a role in the development of T cell-mediated experimental allergic conjunctivitis (EC). EC was induced in Balb/c mice by active immunization with ragweed (RW) followed by RW challenge in eye drops. These mice were treated with neutralizing anti-B7-1 Ab, anti-B7-2 Ab, both Abs, anti-cytotoxic T lymphocyte-associated Ag-4 (CTLA-4) Ab or normal IgGs as controls either during the induction phase or the effector phase. With regard to the induction phase treatment, EC was significantly attenuated when both anti B7-1 and anti-B7-2 Abs were injected. In contrast, anti-CTLA-4 Ab treatment significantly exacerbated EC. With regard to the effector phase treatment, anti B7-2 Ab alone significantly attenuated EC, while anti-CTLA-4 Ab tended to exacerbate EC. Collectively, B7-1 and B7-2 differently contribute to the development of EC during the induction and effector phases. PMID- 17109974 TI - Educational performance of pediatric cochlear implant recipients in mainstream classes. AB - OBJECTIVES: The present study documents the school performance of 20 pediatric cochlear implant recipients who attended mainstream classes and compares their educational performance with their normally hearing peers. METHODOLOGY: All 20 school-aged children who underwent cochlear implantation at the Universiti Kebangsaan Malaysia cochlear implant programme participated in this study. Three measures were employed to assess the school performance. First, using the SIFTER teacher-rating scale, the second measure was the child's examination results, and the third was the child's standing compared to his/her peers in language subject, mathematics, and the overall academic performance during the end of semester examinations. RESULTS: The SIFTER rating scale indicated that only 11.8% of the children were identified as not educationally at risk, 17.6% passed four of the SIFTER subtests, whereas the other 71.6% failed in at least two of the subtests on SIFTER. The highest pass rate was obtained in behavior subtest (76.5%), followed by classroom participation (70.6%), attention (58.8%), academic (47.1%), and communication (11.8%). On the educational performance, the cochlear implant recipients performed significantly better in mathematics (mean scores 62.67%; S.D. 22.24) than in language (mean scores 49.96%, S.D. 25.88) (p<0.01). In the overall examination performance, 25.00% had above average performance (>75th percentile), 18.75% had average performance (25-75th percentile), and another 56.25% performed at below average (<25th percentile). CONCLUSION: Children with cochlear implant were rated poorly in the SIFTER communication subtest. It is possible that language deficit presents an educational challenge in these children. The educational performance of children with cochlear implants in mainstream classes varies. Although 43.75% of them thrive well in a full-time mainstream setting, a significant percentage of them (56.25%) performed at below the average level. These findings reemphasize that although a cochlear implant has successfully provided deaf children with a good hearing potential, the majority of its recipients still require additional educational supports in order to function well in the mainstream educational setting. PMID- 17109975 TI - Highly selective action of triphosphate metabolite of 4'-ethynyl D4T: a novel anti-HIV compound against HIV-1 RT. AB - 2',3'-Didehydro-3'-deoxy-4'-ethynylthymidine (4'-Ed4T), is a recently discovered nucleoside reverse transcriptase inhibitor (NRTI) showing a 5- to 10-fold greater anti-human immunodeficiency virus type 1 (HIV-1) activity and less cellular and mitochondrial toxicity than its parental compound, stavudine (D4T). It is also active against a variety of NRTI-resistant HIV-1 mutants under non-cytotoxic concentrations. In this study, the effects of 4'-Ed4TTP, which is the triphosphate metabolite of 4'-Ed4T, on HIV-1 reverse transcriptase (RT) activity were investigated. We found that 4'-Ed4TTP was a substrate of HIV-1 RT serving as a DNA chain terminator, and it inhibited the DNA polymerase activity of RT more efficiently than D4TTP. The value of Ki(4'-Ed4TTP)/Km(dTTP) is 0.15 for DNA/RNA primer/template duplex (P/T), but 0.7 for DNA/DNA P/T, suggesting 4'-Ed4TTP inhibits RT more efficiently during RNA-dependent DNA synthesis than DNA dependent DNA synthesis. 4'-Ed4TTP was also found to inhibit the 3TC (Lamivudine) resistant RT mutant, M184V, with 3-fold less efficiency than the wild type (wt) RT. 4'-Ed4TTP showed much less inhibitory effects toward major host DNA polymerases. Overall, our results suggest that 4'-Ed4TTP is the active form for anti-HIV-1 activity via its inhibitory effect against RT. PMID- 17109976 TI - Secretory and electrophysiological characteristics of insulin cells from gastrectomized mice: evidence for the existence of insulinotropic agents in the stomach. AB - Mice were subjected to gastrectomy (GX) or sham operation (controls). Four to six weeks later the pancreatic islets were isolated and analysed for cAMP or alternatively incubated in a Krebs-Ringer based medium in an effort to study insulin secretion and cAMP accumulation in response to glucose or the adenylate cyclase activator forskolin. Freshly isolated islets from GX mice had higher cAMP content than islets from control mice, a difference that persisted after incubation for 1 h at a glucose concentration of 4 mmol/l. Addition of forskolin to this medium induced much greater cAMP and insulin responses in islets from GX mice than in islets from control mice. In contrast, the insulin response to high glucose (16.7 mmol/l) was much weaker in GX islets than in control islets. Glucose-induced insulin release was associated with a 2-fold rise in the cAMP content in control islets. Surprisingly no rise in cAMP was noted in GX islets incubated at high glucose. Capacitance measurements conducted on isolated insulin cells from GX mice revealed a much lower exocytotic response to a single 500 ms depolarisation (from -70 mV to zero) than in control insulin cells. Addition of cAMP to the cytosol enhanced the exocytotic response in insulin cells from control mice but not from GX mice. The depolarisation-triggered inward Ca(2+) current in insulin cells from GX mice did not differ from that in control mice, and hence the reduced exocytotic response following GX cannot be ascribed to a decreased Ca(2+) influx. Experiments involving a train of ten 500 ms depolarisations revealed that the exocytotic response was prominent in control insulin cells but modest in GX insulin cells. It seems that cAMP is capable of eliciting insulin release from insulin cells of GX mice only when cAMP is generated in a specific microdomain conceivably through the intervention of membrane-associated adenylate cyclases that can be activated by forskolin. The GX evoked impairment of depolarisation-induced exocytosis and glucose-stimulated insulin release may reflect the lack of a gastric agent that serves to maintain an appropriate insulin response to glucose and an appropriate exocytotic response to depolarisation by raising cAMP in a special glucose-sensitive compartment possibly regulated by a soluble adenylate cyclase. PMID- 17109977 TI - Adiponectin modulates carnitine palmitoyltransferase-1 through AMPK signaling cascade in rat cardiomyocytes. AB - Adiponectin, an adipocyte-derived polypeptide hormone, plays an important role in regulating fatty acid oxidation. beta-oxidation of fatty acids supplies most of the cardiac energy and carnitine palmitoyltransferase (CPT)-1 serves as a key regulator during this process. To characterize the potential effects of adiponectin on CPT-1, we incubated rat neonatal cardiomyocytes with globular adiponectin (gAd). Results showed that gAd promoted the activity and mRNA expression of CPT-1. The underlying signal pathway involved in this modulatory effect was further investigated. Inhibition of AMP-activated protein kinase (AMPK) with adenine 9-beta-d-arabinofuranoside (AraA) completely abrogated gAd mediated AMPK and acetyl coenzyme A carboxylase (ACC) phosphorylation and suppressed the promotion of CPT-1 activity. gAd also induced the phosphorylation of p38 mitogen-activated protein kinase (MAPK) and peroxisome proliferator activated receptor (PPAR)-alpha, which was inhibited by AraA. SB202190, a p38MAPK inhibitor, blocked gAd-stimulated PPAR-alpha phosphorylation. When AMPK and/or p38MAPK was inhibited, gAd-enhanced mRNA expression of CPT-1 was partially reduced. In conclusion, our study suggests that the activation of AMPK signaling cascade participates in the promotion effect of gAd on CPT-1. PMID- 17109978 TI - Acquired noncompaction associated with myopathy. AB - In a 40-year-old woman, hospitalised in May 2001 because of heart failure NYHA IV due to dilated cardiomyopathy, echocardiography had shown a normally sized but poorly contracting left ventricle with normal wall thickness. In October 2005, echocardiography in the same patient revealed a dilated left ventricle with extensive trabeculations and deep intertrabecular recesses in the lateral and posterior wall. The ratio of the noncompacted/compacted myocardium was 2. At clinical neurologic examination myopathy was suspected. Left ventricular hypertrabeculation/noncompaction may be associated with neuromuscular disorders and is not a congenital abnormality in each case. PMID- 17109979 TI - Modifications of composite grafts for coronary bypass surgery. AB - We herein describe four modifications of graft construction in coronary bypass surgery, including composite left mammary arterial T graft, arterial-venous I, and U grafts, and coronary-coronary vein graft and discuss their indications. PMID- 17109980 TI - Serodiagnosis of equine infectious anemia by agar gel immunodiffusion and ELISA using a recombinant p26 viral protein expressed in Escherichia coli as antigen. AB - We used a p26 recombinant protein (p26r) from equine infectious-anemia virus (EIAV) expressed in Escherichia coli as antigen to standardize an agar-gel immunodiffusion (AGIDp26r) test and an indirect ELISA (ELISAp26r) for the detection of antibodies against EIAV in 720 equine sera from Brazil. We evaluated the tests's relative diagnostic sensitivities (relSe) and relative diagnostic specificities (relSp) against a commercial AGID kit (Idexx, USA). We used three sera panels: panel A--196 AGID-negative sera from an AIE non-endemic controlled area; panel B--194 AGID-negative sera from an AIE endemic area and panel C--330 AGID-positive sera from an AIE endemic area. ELISAp26r cut-off value was defined with TG-ROC using sera from panels A and C. AGIDp26r showed an agreement of 100% with the commercial kit. When applied to sera from panels A and C, ELISAp26r showed an agreement of 100% with the kit, but, although relSe was 100% for panel C, the ELISAp26r had relSp of 93.3%. PMID- 17109981 TI - Enhanced production of azadirachtin by hairy root cultures of Azadirachta indica A. Juss by elicitation and media optimization. AB - Azadirachtin is one of the most potent biopesticides so far developed from a plant sources. Influence of different culture media and elicitation on growth and production of azadirachtin by hairy root cultures of Azadirachta indica was studied. Out of the three media tested, namely Ohyama and Nitsch, Gamborg's and Murashige and Skoog's basal media, hairy roots cultured on Ohyama and Nitsch's basal medium produced maximum yield of azadirachtin (0.0166% dry weight, DW). Addition of biotic elicitor enhanced the production of azadirachtin by approximately 5-fold (0.074% DW), while signal compounds such as jasmonic acid and salicylic acid showed a approximately 6 (0.095% DW) and approximately 9-fold (0.14% DW) enhancement, respectively, in the production of azadirachtin as compared to control cultures on Ohyama and Nitsch medium. Extracts from hairy roots were found to be superior to those from the leaves for antifeedant activity against the larvae of Spodoptera litura. PMID- 17109982 TI - Parapoxvirus of red deer in New Zealand encodes a variant of viral vascular endothelial growth factor. AB - Parapoxvirus of red deer in New Zealand (PVNZ), a species of the Parapoxvirus genus, causes scabby lesions on the skin and the velvet of red deer. The three other species of the genus have each been shown to encode homologs of vascular endothelial growth factor (VEGF). We report here that PVNZ strain RD86 also encodes a VEGF and that the predicted PVNZ protein shows only 37-54% amino acid identity to VEGFs encoded by the other species of the genus. Despite this extensive sequence divergence, assays of purified PVNZ VEGF (PVNZ(RD86)VEGF) demonstrated that it shares the unique VEGF receptor (VEGFR) binding profile of the other parapoxvirus VEGFs, in that it binds VEGFR-2 and induces VEGFR-2 mediated proliferation of Ba/F3-derived cells, but does not bind VEGFR-1 or VEGFR 3. In contrast to some other viral VEGFs, it does not bind neuropilin-1. Our results indicate that PVNZ(RD86)VEGF is a biologically active member of the VEGF family and is likely to contribute to the proliferative and highly vascularized nature of PVNZ lesions. Our data also reveal that all members of the genus encode a VEGF and that an extraordinary degree of inter-species sequence variation is a general feature of the parapoxvirus VEGFs. PMID- 17109983 TI - Deletion and recombination events between the DNA-A and DNA-B components of Indian cassava-infecting geminiviruses generate defective molecules in Nicotiana benthamiana. AB - Cloned DNA-B components, belonging to the bipartite begomoviruses Indian cassava mosaic virus (ICMV) and Sri Lankan cassava mosaic virus (SLCMV), family Geminiviridae, when co-inoculated along with previously cloned DNA-A components of the respective viruses onto the experimental host Nicotiana benthamiana, generated defective DNAs (def-DNA) ranging in size from 549 to 1555 nucleotides. All the cloned def-DNAs contained the common region (CR) as well as portions of either DNA-A or DNA-B and, in a few cases, both DNA-A and DNA-B, representing recombinant products, the junction points of which correspond to repeats of 2-11 bases found in the parental molecules. The DNA-B-derived def-DNAs were, in some cases, associated with a decrease in levels of DNA-B, with a concomitant change in the symptoms from downward leaf curling in the older leaves to upward leaf rolling in newly emerging leaves, more typical of monopartite begomoviruses. PMID- 17109984 TI - Strawberry chlorotic fleck: identification and characterization of a novel Closterovirus associated with the disease. AB - Chlorotic fleck, a strawberry disease caused by a graft and aphid transmissible agent, was identified more than 45 years ago in Louisiana. Since its discovery there has been no additional information on the agent that causes the disease. The mode of transmission implies that a virus is the causal agent of chlorotic fleck. We identified four closteroviruses in the single chlorotic fleck infected strawberry clone known to exist in the United States. Sequence analysis indicated that two of the viruses are novel and one of them is closely related to members of the Closterovirus genus, the aphid-transmitted viruses in the family Closteroviridae, a feature that is in accordance with the aphid transmissibility of the chlorotic fleck agent. The genome of the novel Closterovirus, designated as Strawberry chlorotic fleck associated virus exceeds 17 kilobases and encodes 10 open reading frames, including the signature closterovirus genes as well as a gene without obvious homologs in the family. RNA folding predicted a pseudoknot structure near the 3' terminus of the virus that may be involved in template recognition by the viral polymerase. Phylogenetic analysis indicates that Strawberry chlorotic fleck associated virus is most closely related to Citrus tristeza virus among sequenced members of the family. Detection protocols have been developed and the virus was detected in several strawberry plants from production fields. PMID- 17109985 TI - Indomethacin-loaded polymeric nanocarriers based on amphiphilic polyphosphazenes with poly (N-isopropylacrylamide) and ethyl tryptophan as side groups: Preparation, in vitro and in vivo evaluation. AB - The effects of copolymer composition, drug structure and initial drug feed on drug loading of polymeric micelles based on amphiphilic polyphosphazenes were investigated. It was found that the drug loading capacity of micelles based on this type of amphiphilic copolymers was mainly determined by copolymer composition and the chemical structure of drug. In addition to the compatibility between drug and micellar core, hydrogen bonding interaction between drug and hydrophilic corona may significantly influence drug loading as well. In vitro drug release in 0.1 M PBS (pH 7.4) suggested that indomethacin (IND) in the micelles was released through Fickian diffusion, and no significant difference in release rate was observed for micelles based on copolymers with various EtTrp content. Compared with in vitro IND release profile, in vivo pharmacokinetic study after subcutaneous administration provides a more sustained release behavior. Additionally, in comparison with free drug solution at the same dose, IND concentration in rat plasma showed a prolonged retention when the drug was delivered through polymeric micelles. In vivo pharmacodynamic study based on both carrageenan-induced acute and complete Freund's adjuvant-induced adjuvant arthritis model indicated that sustained therapeutic efficacy could be achieved through intraarticular injection of IND-loaded micelles. Most importantly, local delivery of IND can avoid the severe gastrointestinal stimulation, which was frequently associated with oral administration. PMID- 17109986 TI - Hepatic safety profile and glycemic control of pioglitazone in more than 20,000 patients with type 2 diabetes mellitus: postmarketing surveillance study in Japan. AB - The prospective observational study was designed to identify factors affecting glycemic control with pioglitazone and to confirm the hepatic safety of the drug in patients with type 2 diabetes. Baseline patient characteristics, changes in serum hemoglobin A1c (A1c) and alanine aminotransferase (ALT), other treatments for diabetes mellitus, and hepatobiliary adverse reactions were examined. In total, 24,993 patients, representing 28,008 patient-years, were included in the safety evaluation and 20,447 patients in the efficacy evaluation. No case of hepatic failure was reported, and neither temporal nor dose relations were found between pioglitazone and ALT abnormalities. Serum A1c was clearly reduced in patients with baseline body mass index <25 kg/m(2) or baseline fasting immunoreactive insulin <5.0 microU/mL. Among the patients treated for more than 6 months, the change in A1c was -1.0% at 6 months with both monotherapy and combination therapy and remained stable up to 18 months. The overall rate of achievement of A1c<7% in patients with baseline A1c above 7% was 34.1%; notably, the achievement rate of A1c<7% was approximately 30% even in patients with high baseline A1c (mean 8.8%) taking multiple antidiabetic medications, including sulfonylurea, for whom insulin therapy is usually indicated in Japan. PMID- 17109987 TI - Complete eradication of hepatocellular carcinomas by combined vasostatin gene therapy and B7H3-mediated immunotherapy. AB - BACKGROUND/AIMS: B7H3 immunogene therapy is able to completely eradicate tumors when combined with an anti-vascular agent. The aim of this study was to determine whether vasostatin, a potent anti-angiogenic agent, could synergize with B7H3 mediated immunotherapy to combat hepatocellular carcinoma (HCC). METHODS: Vasostatin and B7H3 expression plasmids were constructed, and the in vitro and in vivo expression and anti-angiogenic activity of recombinant vasostatin were measured. The anti-tumor activities of B7H3 and vasostatin alone and in combination were assessed using single and multiple H22 tumor models. RESULTS: Gene transfer of vasostatin inhibited the proliferation of aortic endothelial cells, and angiogenesis in the chorioallantoic membrane assay. Subcutaneous H22 tumors established in BALB/c mice were completely eradicated in response to intratumoral injection of B7H3-expressing plasmids followed 24h later by vasostatin-expressing plasmids. In contrast, neither vasostatin nor B7H3 monotherapy was effective. Gene transfer of vasostatin inhibited tumor angiogenesis and enhanced infiltration of NK cells, whereas B7H3 therapy activated CD8+ and NK cells and increased their infiltration into tumors, and enhanced the levels of circulating IFN-gamma. B7H3 and vasostatin combination gene therapy was effective in combating a systemic challenge of parental H22 cells, and caused the complete regression of multiple distant tumor nodules. CONCLUSIONS: Combining vasostatin anti-angiogenic therapy with B7H3-mediated immunotherapy warrants investigation as a therapeutic strategy to combat HCC, and other malignancies. PMID- 17109988 TI - Governing UK medical performance: a struggle for policy dominance. AB - In the UK, policy on the governance of medical performance is characterised by a continuing struggle between state and profession for control of the agenda setting, formation and implementation stages of the policy process. Since 1998 both sides have continued to produce policies in response to highly visible political pressures but have yet to agree on how those policies should engage as they are implemented at the level of the individual practitioner. For the state, clinical governance forms the lynchpin of its drive to increase managerial control over doctors and, for the profession, revalidation is seen as the means for ensuring the quality of medical performance whilst preserving medicine's historic autonomy. This paper analyses the course of this 7-year struggle and shows how in constructing and delivering policy, state and profession draw on quite different and separate sets of institutional structures and values. As a consequence, there is an unresolved competition for dominance and little engagement between the two policy streams. PMID- 17109989 TI - Understanding SAGE data. AB - Serial analysis of gene expression (SAGE) is a method for identifying and quantifying transcripts from eukaryotic genomes. Since its invention, SAGE has been widely applied to analyzing gene expression in many biological and medical studies. Vast amounts of SAGE data have been collected and more than a thousand SAGE-related studies have been published since the mid-1990s. The principle of SAGE has been developed to address specific issues such as determination of normal gene structure and identification of abnormal genome structural changes. This review focuses on the general features of SAGE data, including the specificity of SAGE tags with respect to their original transcripts, the quantitative nature of SAGE data for differentially expressed genes, the reproducibility, the comparability of SAGE with microarray and the future potential of SAGE. Understanding these basic features should aid the proper interpretation of SAGE data to address biological and medical questions. PMID- 17109990 TI - I am what I eat and I eat what I am: acquisition of bacterial genes by giant viruses. AB - Giant viruses are nucleocytoplasmic large DNA viruses (NCLDVs) that infect algae (phycodnaviruses) and amoebae (Mimivirus). We report an unexpected abundance in these giant viruses of islands of bacterial-type genes, including apparently intact prokaryotic mobile genetic elements, and hypothesize that NCLDV genomes undergo successive accretions of bacterial genes. The viruses could acquire bacterial genes within their bacteria-feeding eukaryotic hosts, and we suggest that such acquisition is driven by the intimate coupling of recombination and replication in NCLDVs. PMID- 17109991 TI - Projective filtering of time-aligned ECG beats for repolarization duration measurement. AB - Automatic measurement of ventricular repolarization duration is difficult and highly prone to errors. We propose to mitigate this problem by applying projective filtering to enhance the ECG signal prior to measurements. Our method performs time-alignment of ECG beats and then employs the rules of principal component analysis to the desired ECG reconstruction. As a result, significant noise reduction is obtained while distortions of the desired ECG components remain negligible. The proposed method is presented under white noise conditions; the influence on the precision of repolarization duration measurement is then investigated. Finally, the method's influence on evaluation of repolarization duration variability when the analyzed signals are contaminated with real electromyographic noise is discussed. PMID- 17109992 TI - Comparative genomic hybridization array analysis and real time PCR reveals genomic alterations in squamous cell carcinomas of the lung. AB - Genomic alterations have been identified in lung cancer tissues and reported in numerous studies. To analyze genomic aberrations in lung cancer patients, we used array comparative genomic hybridization (array CGH) in 14 squamous cell lung carcinoma (SqC) tissues. Copy number gain and loss in chromosomal regions were detected, and the corresponding genes were confirmed by real time PCR. Several frequently altered loci, including gain of 3q (36% of samples), were found. The most frequently identified losses were found at 14q32.33 (21% of samples). The relative degree of chromosomal change was analyzed using log2 ratios. High-level DNA amplifications (>0.8 log2 ratio) were detected at 20 regions in 1p, 2q, 3q, 4q, 6q, 7p, 8q, 9p, 10q, 12q, 14q and 19p. We found that the fold change levels were highest at EVI1 (3q26.2), LPP (3q27-28) and FHF-1 (3q28) gene loci. Our results show that array CGH is a useful tool for identification of gene alteration in lung cancer, and that the above-mentioned genes might represent potential candidate genes for pathogenesis and diagnosis of lung cancer. PMID- 17109993 TI - Gadolinium deposition in nephrogenic fibrosing dermopathy. AB - There is growing recognition of the association between the use of gadolinium containing radiocontrast agents for magnetic resonance imaging and the serious dermal and systemic disease nephrogenic fibrosing dermopathy/nephrogenic systemic fibrosis (NFD/NSF). The pathogenesis of this entity remains unclear; however, our recent observations suggest a likely mechanism for the initial dermal manifestations of this gadolinium toxicity. PMID- 17109994 TI - Neutrophilic eccrine hidradenitis masquerading as facial cellulitis. AB - Neutrophilic eccrine hidradenitis typically manifests as erythematous plaques on the face, trunk, or extremities. This eruption has been associated with numerous factors, but most commonly is seen with chemotherapy, particularly cytarabine. We report a 73-year-old woman with acute myelogenous leukemia who developed rapidly expansive neutrophilic eccrine hidradenitis mimicking facial cellulitis only after a course of cytarabine was followed by granulocyte-colony stimulating factor. Prompt diagnosis is imperative to prevent prolonged antimicrobial therapy. PMID- 17109995 TI - Evaluation of digital dermoscopy in a pigmented lesion clinic: clinician versus computer assessment of malignancy risk. AB - Digital dermoscopy systems employ computer-based algorithms to quantitate features of pigmented skin lesions (PSLs) and provide an assessment of malignancy risk. We evaluated interobserver concordance of PSL malignancy risk between a pigmented lesion specialist and an artificial neural network (ANN)-based automated digital dermoscopy system. While digital dermoscopy provides a reliable means of image capture, storage, and comparison of PSLs over time, the ANN algorithm requires further training and validation before the malignancy risk assessment feature can be widely used in clinical practice. PMID- 17109996 TI - Explaining school children's fruit and vegetable consumption: the contributions of availability, accessibility, exposure, parental consumption and habit in addition to psychosocial factors. AB - We studied the contributions of parental fruit and vegetable (F&V) consumption, availability and accessibility of F&V in the home, exposure to F&V, and habit, in addition to psychosocial factors, in explaining F&V consumption in 4-12-year-old children. Furthermore, we looked for effect modification by ethnicity and gender. Children's parents (n = 1739) completed a questionnaire assessing psychosocial and additional factors regarding their children's F&V consumption. Consumption was assessed by a food-frequency questionnaire. The model explained the children's F&V consumption better when the additional factors were included (R2 = .49 and R2 = .50 for fruit consumption, and R2 = .33 and R2 = .33 for vegetable consumption). Stepwise multi-level regression analyses revealed that habit was the most influential correlate of F&V consumption. It is concluded that nutrition education interventions aimed at stimulating F&V consumption among children should take into account that the consumption of fruit and that of vegetables are clearly different behaviors, with different influencing factors for boys and girls and children of native or non-native background. Furthermore, interventions to increase F&V consumption should include strategies aimed at making these behaviors habitual. PMID- 17109997 TI - Apelin stimulates proliferation and suppresses apoptosis of mouse osteoblastic cell line MC3T3-E1 via JNK and PI3-K/Akt signaling pathways. AB - The aim of this study was to investigate the effects of apelin on proliferation and apoptosis of mouse osteoblastic MC3T3-E1 cells. APJ was expressed in MC3T3-E1 cells. Apelin did not affect Runx2 expression, alkaline phosphatase (ALP) activity, osteocalcin and type I collagen secretion, suggesting that it has no effect on osteoblastic differentiation of MC3T3-E1 cells. However, apelin stimulated MC3T3-E1 cell proliferation and inhibited cell apoptosis induced by serum deprivation. Our study also shows that apelin decreased cytochrome c release and caspase-3, capase-8 and caspase-9 activation in serum-deprived MC3T3 E1 cells. Apelin activated c-Jun N-terminal kinase (JNK) and Akt (phosphatidylinositol 3-kinase downstream effector), and the JNK inhibitor SP600125, the phosphatidylinositol 3-kinase (PI3-K) inhibitor LY294002 or the Akt inhibitor 1L-6-hydroxymethyl-chiro-inositol 2-(R)-2-O-methyl-3-O octadecylcarbonate (HIMO) inhibited its effects on proliferation and serum deprivation-induced apoptosis. Furthermore, apelin protected against apoptosis induced by the glucocorticoid dexamethasone or TNF-alpha. Apelin stimulates proliferation and suppresses serum deprivation-induced apoptosis of MC3T3-E1 cells and these actions are mediated via JNK and PI3-K/Akt signaling pathways. PMID- 17109998 TI - Continuing education in nursing: a concept analysis. AB - The importance of continuing education for nurses has been increasingly emphasized in the nursing literature since the beginning of the profession. The concept of continuing education is often used as a substitute for associated terms such as continuing professional development and lifelong learning, thus highlighting a need for its clarification. The purpose of this article is to explain and describe continuing education, in order to encourage a broader understanding of the concept among nurses. The concept analysis is directed by Rodgers' [Rodgers, B.L., 1989. Concept analysis and the development of nursing knowledge: the evolutionary cycle. Journal of Advanced Nursing 14, 330-335] 'evolutionary approach' which is viewed as an ongoing dynamic process, and one that identifies the shared meaning of concepts. Examining everyday discourse used in the nursing literature identified the critical attributes, antecedents and consequence of continuing education in nursing. As a result, the emerging attributes of the concept are synthesised into a conceptual model. The article concludes with an exploration of the application of the concept of continuing education within nursing and its implications for professional development. PMID- 17109999 TI - Nurses' experiences of continuing professional development. AB - Continuing professional development is regarded as part of the nursing role in the National Health Service. Health policy in the United Kingdom is built on the assumption that nurses' roles can be extended through continuing professional development which is also considered to be a key factor in nursing retention. Previous research has considered the provision of learning mainly from the perspective of managers and education providers. The purpose of this paper is to explore nurses' experiences of continuing professional development. The results presented in this paper form part of a larger survey (n=451, response rate=64.9%) designed to examine opportunities for continuing professional development and factors encouraging and discouraging uptake. An unexpectedly high number of respondents (n=125, 27.7%) commented extensively on their personal experiences of continuing professional development. These comments were analysed inductively. Five main themes emerged: who and what is continuing professional development for?; accessing continuing professional development; one size does not fit all; managing work, life and doing continuing professional development; and making the best of continuing professional development. Most responses were positive, but the demands of taking courses were perceived to encroach on life outside work, especially the need to complete lengthy assignments. Managers were perceived to operate as the gatekeepers to course admission and opportunities to implement new knowledge and expertise. No comments were received concerning the role of education staff in continuing professional development. PMID- 17110000 TI - Cross-reactive immune responses in mice after genetic vaccination with cDNA encoding hantavirus nucleocapsid proteins. AB - Hantaviruses cause hemorrhagic fever with renal syndrome (HFRS) in about 150,000 individuals in Eurasia, and several hundred cases of hantavirus pulmonary syndrome (HPS) on the American continent annually. There is consequently a need for rapid diagnostics and effective prevention of hantaviral infections. In this study we have performed DNA-vaccination of mice with full-length genes encoding the immunogenic nucleocapsid protein (NP) of Puumala (PUUV), Seoul (SEOV) and Sin Nombre virus (SNV). The antibody reactivity towards the NPs, and deleted or truncated variants thereof, were studied to localise and investigate the major polyclonal B-cell epitopes. Our findings clearly show that the antibody reactivity in each immunised mouse is unique, not only in a quantitative respect (titers) but also in cross-reactivity and most likely also in the epitope specificity. Our experimental data in combination with B-cell prediction software indicate that strong homologous virus species specific and cross-reactive epitopes are located around amino acid residue 40 in the nucleocapsid proteins. PMID- 17110001 TI - Comparison of mucosal and parenteral immunisation in two animal models of pneumococcal infection: otitis media and acute pneumonia. AB - Bacterial clearance and immune responses in a mouse model of pneumonia and a rat model of otitis media following parenteral or mucosal immunisation in both models were compared. Both the immunisation routes were equally effective in inducing bacterial clearance from the lung, upregulated the recruitment of white cells and lead to an increase in the concentration of TNF-alpha, IL-1beta and specific antibody in the bronchoalveolar lavage. Both the routes of immunisation enhanced clearance of bacteria from the middle ear. Parenteral immunisation was most effective overall in enhancing bacterial clearance and recruiting white cells to the middle ear. Both the routes significantly suppressed the levels of TNF-alpha in the middle ear lavage. Mucosal immunisation induced a Streptococcus pneumoniae specific IgA antibody response. Both the animal models gave highly reproducible disease and provided high levels of sensitivity for testing the efficacy of candidate vaccine antigens. Differences observed in the inflammatory responses require further study. PMID- 17110002 TI - Can ECOPATH with ECOSIM enhance models of radionuclide flows in food webs? An example for 14C in a coastal food web in the Baltic Sea. AB - In this study it was evaluated whether the ECOPATH with ECOSIM software could be used as a platform to facilitate the construction of models and study of transport and accumulation of radionuclides in aquatic food webs. The evaluation was based upon a food web model of carbon (C) and carbon-14 ((14)C) flow for a coastal area in the Baltic Sea, the ECOPATH, the ECOSIM and the ECOTRACE models. The original carbon flows and assumptions were easily incorporated into the ECOPATH and ECOSIM modelling environment. The new model was also well suited to drive a (14)C flow model (ECOTRACE) for each of the organisms included. ECOTRACE estimated steady-state concentrations of (14)C that were between 73 and 142% of the original flows. The results clearly show that there is great potential for a successful development of this approach for integrating scientific knowledge about food webs and radioecological models for aquatic systems. PMID- 17110003 TI - Sediment radioisotope dating across a stratigraphic discontinuity in a mining impacted lake. AB - Application of radioisotope sediment dating models to lakes subjected to large anthropogenic sediment inputs can be problematic. As a result of copper mining activities, Torch Lake received large volumes of sediment, the characteristics of which were dramatically different from those of the native sediment. Commonly used dating models (CIC-CSR, CRS) were applied to Torch Lake, but assumptions of these methods are violated, rendering sediment geochronologies inaccurate. A modification was made to the CRS model, utilizing a distinct horizon separating mining from post-mining sediment to differentiate between two focusing regimes. (210)Pb inventories in post-mining sediment were adjusted to correspond to those in mining-era sediment, and a sediment geochronology was established and verified using independent markers in (137)Cs accumulation profiles and core X-rays. PMID- 17110004 TI - Pedicled genial osteotomy modification of the mandibular release access operation for access to the back of the tongue. AB - We report a modification of the operation of mandibular release or 'visor drop down'. It allows more accurate repositioning and more permanent fixation of the genioglossus, geniohyoid, and digastric muscles using a small anterior osteotomy. We hope that this more accurate and reliable repositioning will give improved outcomes in terms of speech, chewing, and swallowing. PMID- 17110005 TI - Prophylactic internal fixation of the radial osteocutaneous donor site. AB - The technique of prophylactic internal fixation (PIF) of the radial osteocutaneous donor site is described and reviewed. Twenty-two donor sites were reinforced with a 3.5mm dynamic compression plate across the anterior defect. The incidence of fracture was 4.5% (1 out of 22). The single fracture was due to a technical error and was managed conservatively. Fracture at a donor site that has not been plated is more frequent and often becomes displaced, requiring secondary surgery. In contrast, the incidence of fracture, displacement and secondary surgery following prophylactic internal fixation (PIF) is relatively low. There have been no significant long-term complications with PIF. It is now the method of choice for managing the radial osteocutaneous donor site. PMID- 17110006 TI - One miniplate versus two in the management of mandibular angle fractures: a prospective randomised study. AB - We compared the use of one miniplate (n=36) with that of two miniplates (n=26) for the treatment of the mandibular angle in a randomised trial. There were no significant differences between the groups in total morbidity (22/36 compared with 14/26) or for individual complications. We conclude that two miniplates seem to confer no extra benefit to patients, but a much larger trial would be required to show this conclusively. PMID- 17110007 TI - Differential effects of cyclic and static pressure on biochemical and morphological properties of chondrocytes from articular cartilage. AB - BACKGROUND: Mechanical stresses are known to play important role on articular cartilage functions in vivo and also on cartilage explants and chondrocytes monolayer culture. This study examined the differential effect of cyclic and static pressures on chondrocytes cultured in alginate matrix, which is physiologically closer to the in vivo environment of cells in cartilage. METHODS: Goat knee joint articular cartilage chondrocytes cultured in alginate beads were exposed to 1.2 and 2.4 MPa cyclic and static loadings via a custom-made cam/follower based machine. Biochemical contents (glycosaminoglycan, collagen, DNA) and protease activity of cells were analyzed separately in cellular matrix, further removed matrix and in culture medium. Morphology of chondrocytes was studied under transmission electron microscopy. FINDINGS: Compared with controls (unloaded cells), cyclic loading increased the glycosaminoglycan content of cells at 1.2 and 2.4 MPa in cellular matrix and further removed matrix (P<0.001) whereas it decreased at similar static loads (P<0.001). In alginate matrix, chondrocytes released a metalloprotease, which required Mn(2+) for activity. Both cyclic load levels inhibited its specific activity in cellular matrix but increased it at static loading (P<0.001). The protease specific activity in further removed matrix increased at both cyclic and static loadings (P<0.001). Transmission electron microscopy data showed improved cells ultrastructure and cell-matrix interactions under cyclic load whereas these deteriorated under static loadings. INTERPRETATION: The study suggests that cyclic load has a positive effect on chondrocytes metabolism and morphology whereas static load has a degenerative effect. PMID- 17110008 TI - Concern and practice among men about HIV/AIDS in low socioeconomic income areas of Lilongwe, Malawi. AB - The HIV prevalence rate in Malawi, currently estimated to be 15%, is among the highest in the world. There is a growing realization that in order to understand the underlying causes and devise more effective prevention strategies focus should be placed on economic, political, social, and cultural forces as well as perceptions of individual risk to HIV/AIDS. During 2003 we conducted field work in Lilongwe, the capital city of Malawi, to examine perceptions of individuals as to their risk to HIV infection using the structured interviews and the focus group discussions with men from five areas of Lilongwe. The discussion in this paper focuses on the perception of risk to HIV infection among men in low socioeconomic income areas that we interviewed. Our findings indicate that while knowledge about HIV/AIDS and the best ways in which one can protect oneself from getting HIV is very high, people continue to engage in at-risk behaviors without using the necessary protection. Many of the men in our sample indicated that they were indeed at risk of getting infected with HIV. In spite of this, some of the respondents in both the structured interviews and the focus group discussions pointed out that some people had began taking measures to protect themselves, such as using condoms with nonregular partners, women leaving their husbands where cheating was obvious, and, for men, reducing the number of extra-marital sexual relations. PMID- 17110009 TI - Developing, integrating, and perpetuating new ways of applying sociology to health, medicine, policy, and everyday life. AB - As a framework for presenting ideas on developing ways to make sociology more applicable, we focus on the recent state of medical sociology research. Data for this paper were generated through a content analysis of a twelve-year period (1993-2004) of the Journal of Health and Social Behavior (JHSB) and Social Science & Medicine (SSM). The analysis aims to determine if the content of JHSB and SSM reflect the breadth of the sub-discipline of medical sociology as well as the stated goals of the journals. The selected issues of JHSB and SSM were coded on the basis of the following attributes: (1) Primary Substantive Topic, (2) Methodology, (3) Data Type and Analytic Technique, and (4) Research and Policy Recommendations. We found that the orientation of JHSB articles was towards generating research and theory that shy away from policy, interdisciplinary approaches, and applied issues. SSM content tends to display more interdisciplinary breadth and variety, but also reflects a dearth of applied recommendations. Our discussion focuses on what JHSB and SSM could be. We present ideas on how the sociological discipline in general-and JHSB and SSM in particular-can help generate and nourish new forms of inquiry that can impact the way research questions are framed. We conclude that such a shift is needed in order to maximize the applicability of social scientific evidence to everyday life, and we share examples situated within a socio-medical context, where there is a particular need for the application of social evidence to practice. PMID- 17110010 TI - Prolactin and estradiol serum levels in unmedicated male paranoid schizophrenia patients. AB - There is evidence for the involvement of the endocrine system in schizophrenia. This involment was widely investigated in female patients. In the current study, we examined prolactin and estradiol serum levels in hospitalized unmedicated men with first-episode and recurrent schizophrenia and then tested possible correlation with various subtypes of the disease. In addition, the estradiol and prolactin levels were compared with a healthy control group. The serum samples were assessed the morning following admission in fifty-seven schizophrenia male patients. There was a significant difference in prolactin serum levels between the paranoid and "nonparanoid" schizophrenia subgroups. However, no significant differences were found in estradiol serum levels between schizophrenia subtypes or between the patients and their healthy counterparts. Finally, a significant and positive correlation was found between the prolactin and estradiol levels in the paranoid subgroup alone. Thus, it appears that low estradiol levels are associated with low prolactin levels, alleged hyperdopaminergic tone and psychotic breakdown in paranoid schizophrenia. The results of the present study further support our previous report of the association between prolactin serum levels and the schizophrenia cluster subtypes, indicating a different dopaminergic activity for the various forms of the disease. PMID- 17110011 TI - Use of long-acting injectable risperidone before and throughout pregnancy in schizophrenia. AB - Data on the use of long-acting injectable (LAI) risperidone, the first atypical depot antipsychotic, during pregnancy are limited. A 35-year-old woman with schizophrenia was given LAI risperidone before and throughout her pregnancy. She gave birth to a female infant weighing 2230 g at 36 weeks and 6 days of pregnancy, following premature rupture of the membranes. The baby had no congenital malformation and was healthy 8 months postnatal. To our knowledge, this is the first reported use of LAI risperidone throughout an entire pregnancy. In this paper, we discuss the rationale and problems of LAI risperidone use in pregnancy, based on a literature review. PMID- 17110012 TI - Problems in the supply of antidotes--a view from the pharmaceutical industry. AB - Antidotes are drugs and therefore subject to numerous legal requirements. Due to their indication they are usually needed in small quantities. This is to be considered in view of the efforts to comply with the regulatory affairs. The production capacity and the kept in stock quantities in the pharmaceutical industry for economical reasons can only be small. In cases of catastrophic large events large quantities of antidotes are needed within short time. In these cases only a national stockpiling program can secure the supply with the necessary medicaments for the citizens. PMID- 17110013 TI - Evaluation of oxidative stress in D-serine induced nephrotoxicity. AB - It has been suggested that oxidative stress is involved in d-serine-induced nephrotoxicity. The purpose of this study was to assess if oxidative stress is involved in this experimental model using several approaches including (a) the determination of several markers of oxidative stress and the activity of some antioxidant enzymes in kidney and (b) the use of compounds with antioxidant or prooxidant effects. Rats were sacrificed at several periods of time (from 3 to 24h) after a single i.p. injection of d-serine (400mg/kg). Control rats were injected with l-serine (400mg/kg) and sacrificed 24h after. The following markers were used to assess the temporal aspects of renal damage: (a) urea nitrogen (BUN) and creatinine in blood serum, (b) kidney injury molecule (KIM-1) mRNA levels, and (c) tubular necrotic damage. In addition, creatinine clearance, proteinuria, and urinary excretion of N-acetyl-beta-d-glucosaminidase (NAG) were measured 24h after d-serine injection. Protein carbonyl content, malondialdehyde (MDA), 4 hydroxy-2-nonenal (4-HNE), fluorescent products of lipid peroxidation, reactive oxygen species (ROS), glutathione (GSH) content, and heme oxygenase-1 (HO-1) expression were measured as markers of oxidative stress in the kidney. Additional experiments were performed using the following compounds with antioxidant or pro oxidant effects before d-serine injection: (a) alpha-phenyl-tert-butyl-nitrone (PBN), a spin trapping agent; (b) 5,10,15,20-tetrakis (4-sulfonatophenyl) porphyrinato iron(III) (FeTPPS), a soluble complex able to metabolize peroxynitrite; (c) aminotriazole (ATZ), a catalase (CAT) inhibitor; (d) stannous chloride (SnCl(2)), an HO-1 inductor; (e) tin mesoporphyrin (SnMP), an HO inhibitor. In the time-course study, serum creatinine and BUN increased significantly on 15-24 and 20-24h, respectively, and KIM-1 mRNA levels increased significantly on 6-24h. Histological analyses revealed tubular necrosis at 12h. The activity of antioxidant enzymes catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase remained unchanged at all times studied. Protein carbonyl content, MDA, 4-HNE, and ROS remained unchanged at all time points studied. GSH content decreased transiently on 9 and 12h. Interestingly, fluorescent products of lipid peroxidation decreased significantly on 3-24h. HO-1 expression was undetectable by Western blot and the immunohistochemistry studies revealed that the intensity of HO-1 staining was weak. The administration of PBN, FeTPPS, ATZ, SnCl(2), and SnMP did not prevent or enhance renal damage induced by d-serine. Our data taken as a whole suggest that oxidative stress is not involved in the early phase of the nephrotoxicity induced by d-serine. PMID- 17110014 TI - Proteolytic processing by matrix metalloproteinases and phosphorylation by protein kinase CK2 of fetuin-A, the major globulin of fetal calf serum. AB - Bovine fetuin-A is a member of a glycoprotein family with a wide spectrum of functions. Until now the bovine protein has been thought to be a single-chain protein. Recently we have shown that native bovine plasma fetuin-A partially exists as a disulfide-bridged two-chain protein with a heavy N-terminal and a lighter C-terminal chain similar to the structure of human fetuin-A homologue (alpha2HS glycoprotein), and also is partially phosphorylated at residues Ser120, Ser302, Ser305 and Ser306 (Wind et al., Anal. Biochem. 317 (2003) 26-33). Both fetuin-A modifications, the phosphorylation at the four sites as well as the proteolysis which causes longer or shorter light chains (termed lc-1 and lc-2, respectively), are probably brought about by targeted enzymatic activities which still need to be defined. In this study we show that authentic bovine fetuin-A disulfide-bridged two-chain forms, which include the original C-terminus, were liberated from the single-chain precursor by metalloproteinases MMP-3 (stromelysin-1) and MMP-7 (matrilysin), but not by elastase, cathepsin E and cathepsin G. Peptide sequencing suggested cleavage sites chiefly at the Pro277 Ser278 or Arg294-His295 peptide bonds. Fetuin-A radioactive phosphorylation in vitro by protein kinase CK2 caused (32)P incorporation into the fetuin-A light chain lc-1 but not lc-2 or the fetuin-A heavy chain, as revealed by MMP assisted proteolysis. Analysis by nanoESI-MS pinpointed phosphorylation at the native phospho-residues Ser302, Ser305 and Ser306 by increased relative abundance following in vitro phosphorylation. Moreover, CK2 phosphorylation of synthetic C terminal fetuin-A peptides, used as effective controls to the native protein, strongly implies that CK2 is involved in the in vivo phosphorylation of fetuin-A. The phosphorylation of N-terminally truncated peptide homologs seemed highly dependent on the sequence context N-terminal of the phosphorylation sites, thus providing a likely explanation for the non-phosphorylation of the light chain lc 2 in native fetuin-A. PMID- 17110015 TI - Purification and partial characterization of two phospholipases A2 from Bothrops leucurus (white-tailed-jararaca) snake venom. AB - Two proteins with phospholipase A(2) (PLA(2)) activity were purified to homogeneity from Bothrops leucurus (white-tailed-jararaca) snake venom through three chromatographic steps: Conventional gel filtration on Sephacryl S-200, ion exchange on Q-Sepharose and reverse phase on Vydac C4 HPLC column. The molecular mass for both enzymes was estimated to be approximately 14 kDa by SDS-PAGE. The N terminal sequences (48 residues) show that one enzyme presents lysine at position 48 and the other an aspartic acid in this position, and therefore they were designated blK-PLA(2) and blD-PLA(2) respectively. blK-PLA(2) presented negligible levels of PLA(2) activity as compared to that of blD-PLA(2). The PLA(2) activity of both enzymes is Ca(2+)-dependent. blD-PLA(2) did not have any effect upon platelet aggregation induced by arachidonic acid, ADP or collagen, but strongly inhibits coagulation and is able to stimulate Ehrlich tumor growth but not angiogenesis. PMID- 17110016 TI - Analysis and prediction of flow from local source in a river basin using a Neuro fuzzy modeling tool. AB - Traditionally, the multiple linear regression technique has been one of the most widely used models in simulating hydrological time series. However, when the nonlinear phenomenon is significant, the multiple linear will fail to develop an appropriate predictive model. Recently, neuro-fuzzy systems have gained much popularity for calibrating the nonlinear relationships. This study evaluated the potential of a neuro-fuzzy system as an alternative to the traditional statistical regression technique for the purpose of predicting flow from a local source in a river basin. The effectiveness of the proposed identification technique was demonstrated through a simulation study of the river flow time series of the Citarum River in Indonesia. Furthermore, in order to provide the uncertainty associated with the estimation of river flow, a Monte Carlo simulation was performed. As a comparison, a multiple linear regression analysis that was being used by the Citarum River Authority was also examined using various statistical indices. The simulation results using 95% confidence intervals indicated that the neuro-fuzzy model consistently underestimated the magnitude of high flow while the low and medium flow magnitudes were estimated closer to the observed data. The comparison of the prediction accuracy of the neuro-fuzzy and linear regression methods indicated that the neuro-fuzzy approach was more accurate in predicting river flow dynamics. The neuro-fuzzy model was able to improve the root mean square error (RMSE) and mean absolute percentage error (MAPE) values of the multiple linear regression forecasts by about 13.52% and 10.73%, respectively. Considering its simplicity and efficiency, the neuro fuzzy model is recommended as an alternative tool for modeling of flow dynamics in the study area. PMID- 17110017 TI - Ranking without valuing in the face of major uncertainty--the case of the promotion of biodegradable lubricants. AB - Major benefit uncertainties prevent monetary quantification of some environmental amenities. Replacing mineral lubricants with biodegradable substitutes is shown to be a case in point. However, it is possible to rank the social benefits of substituting mineral lubricants with regard to different applications and environments. As the private costs and benefits of substitution are mainly constant, only a policy that prioritizes full substitution in the applications with the greatest benefits can be efficient. It is shown that regulations requiring substitution in certain fields are likely to fulfil this criterion, while subsidies for production, processing, market introduction and research usually fail to meet the efficiency criterion. PMID- 17110018 TI - Biomass production and carbon sequestration potential in poplar plantations with different management patterns. AB - Biomass production and carbon storage in short-rotation poplar plantations over 10 years were evaluated at the Hanyuan Forestry Farm, Baoying County, China. Experimental treatments applied in a split-plot design included four planting densities (1111, 833, 625 and 500 stems ha(-1)) and three poplar clones (NL 80351, I-69 and I-72). Based on the model of total biomass production developed, total plantation biomass production was significantly different in the plantations. The ranking of the plantation biomass production by planting density was 1111>833 more more than 625>500 stems ha(-1), and by components was stem>root>or=branch>leaf for all plantations. At 10 years, the highest total biomass in the plantation of 1111 stems ha(-1) reached about 146 t ha(-1), which was 5.3%, 11.6% and 24.2% higher than the plantations of 833, 625 and 500 stems ha(-1), respectively. The annual increment of biomass production over 10 years differed significantly among initial planting densities and stand ages (p<0.01), but no significant difference was observed from age 7 to 10. Mean carbon concentration among all biomass components ranged from 42-50%, with the highest carbon concentrations in stems and the lowest in leaves. Over the study period, the dynamic pattern of total plantation carbon storage by planting density was similar to that of total biomass production. At age 10, the highest total plantation carbon storage in the plantation of 1111 stems ha(-1) reached about 72.0 t ha(-1), which was 5.4%, 11.9% and 24.8% higher than in the plantations of 833, 625 and 500 stems ha(-1), respectively. The annual carbon storage increment over 10 years differed significantly among initial planting densities and stand ages (p<0.01), and it showed a pattern similar to the annual biomass production increment of the plantations. The results suggest that biomass production and carbon storage potential were highest for planting densities of 1111 and 833 stems ha(-1) grown over 5- and 6-year cutting cycles, respectively. If 3- or 4 year cutting cycles are used, the planting density should be higher than 1111 stems ha(-1) (e.g., 1667 or 2500 stems ha(-1)). Based on the mean annual carbon storage for the plantation of 625 stems ha(-1), as an estimation, the mean carbon storage in the biomass of poplar plantations (excluding leaves) amounts to 3.75x10(7) t ha(-1)yr(-1) in China. PMID- 17110019 TI - Ecological footprint analysis applied to a sub-national area: the case of the Province of Siena (Italy). AB - This work is part of a larger project, which aims at investigating the environmental sustainability of the Province of Siena and of its communes, by means of different indicators and methods of analysis. The research presented in this article uses ecological footprint and biocapacity as indicators to monitor the environmental conditions of the area of Siena, thus complementing previous studies carried out using Emergy, greenhouse gases balance and other methods. The calculations have been performed in such a way as to enable a disaggregation of the final results according to the classical categories of ecologically productive land and of consumption, but also according to citizen's and public administration's areas of influence. This information allows us to investigate in detail the socio-economic aspects of environmental resource use. Among the notable results, the Siena territory is characterized by a nearly breakeven total ecological balance, a result contrasting with the national average and most of the other Italian provinces. Furthermore, the analysis has been carried out at different spatial scales (province, districts and communes), highlighting an inhomogeneous territorial structure consisting of subareas in ecological deficit compensated by zones in ecological surplus. PMID- 17110020 TI - Laparoscopic partial nephrectomy for tumour excision in a horseshoe kidney. AB - A 62-yr-old woman with a 2-cm enhancing renal mass in the right moiety of a horseshoe kidney was treated via a four-port transperitoneal laparoscopic approach. The tumour was excised by cold scissors, and the parenchyma was sutured with Vicryl 1 sutures with Surgicel bolster and by using pledgets of Hem-o lok((R)) clips. To our knowledge, this is the first report of a partial nephrectomy for tumour excision on a horseshoe kidney by a transperitoneal laparoscopic approach. PMID- 17110021 TI - One-year follow-up of tension-free vaginal tape (TVT) and trans-obturator suburethral tape from inside to outside (TVT-O) for surgical treatment of female stress urinary incontinence: a prospective randomised trial. AB - OBJECTIVES: To compare tension-free vaginal tape (TVT) and trans-obturator suburethral tape from inside to outside (TVT-O) for surgical treatment of stress urinary incontinence (SUI) for complications (primary end point) and success rate (secondary end point). METHODS: Seventy-two consecutive patients, with a mean age of 53.2 yr (range: 38-69 yr) and affected by SUI, were included in this randomised controlled trial. After preoperative assessment, patients were randomly allocated to the TVT or TVT-O procedure. Operative time, perioperative complications, and hospital stay were prospectively recorded. Cure of SUI was defined as no leakage of urine during the stress test at urodynamic testing at the 12-mo evaluation. The Wilcoxon signed rank sum test, Mann-Whitney U test, McNemar test, and Fisher exact test were used to verify statistical significance, set at p<0.05. RESULTS: All patients were evaluable at the 12-mo follow-up. The characteristics of patients were well balanced between groups after randomisation. The mean operative time was significantly shorter in the TVT-O group. Perioperative complications were significantly more common after the retropubic approach (5% and 27% in TVT-O and TVT groups, respectively, p<0.04). The groups did not differ significantly in intraoperative blood loss, hospital stays, and time to return to normal activities. Sixty-five patients (90%) were successfully treated for SUI 12 mo after the operation (89% and 91% for TVT-O and TVT groups, respectively). CONCLUSIONS: Both techniques appear to be equally effective in the surgical treatment of SUI. However, TVT-O had a shorter operative time and lower overall perioperative complication rate. PMID- 17110022 TI - Novel analytical reagent for the application of cloud-point preconcentration and flame atomic absorption spectrometric determination of nickel in natural water samples. AB - Cloud-point extraction was applied as a preconcentration of nickel after formation of complex with newly synthesized N-quino[8,7-b]azin-5-yl 2,3,5,6,8,9,11,12octahydrobenzo[b][1,4,7,10,13]pentaoxacyclopentadecin-15-yl methanimine, and later determined by flame atomic absorption spectrometry (FAAS) using octyl phenoxy polyethoxy ethanol (Triton X-114) as surfactant. Nickel was complexed with N-quino[8,7-b]azin-5-yl 2,3,5,6,8,9,11,12octahydrobenzo[b][1,4,7,10,13]pentaoxacyclopentadecin-15-yl methanimine in an aqueous phase and was kept for 15 min in a thermo-stated bath at 40 degrees C. Separation of the two phases was accomplished by centrifugation for 15 min at 4000 rpm. The chemical variables affecting the cloud-point extraction were evaluated, optimized and successfully applied to the nickel determination in various water samples. Under the optimized conditions, the preconcentration system of 100 ml sample permitted an enhancement factor of 50 fold. The detailed study of various interferences made the method more selective. The detection limits obtained under optimal condition was 0.042 ngml(-1). The extraction efficiency was investigated at different nickel concentrations (20-80 ngml(-1)) and good recoveries (99.05-99.93%) were obtained using present method. The proposed method has been applied successfully for the determination of nickel in various water samples and compared with reported method in terms of Student's t-test and variance ratio f-test which indicate the significance of present method over reported and spectrophotometric methods at 95% confidence level. PMID- 17110023 TI - Simultaneous removal of organic contaminants and heavy metals from kaolin using an upward electrokinetic soil remediation process. AB - Kaolins contaminated with heavy metals, Cu and Pb, and organic compounds, p xylene and phenanthrene, were treated with an upward electrokinetic soil remediation (UESR) process. The effects of current density, cathode chamber flushing fluid, treatment duration, reactor size, and the type of contaminants under the vertical non-uniform electric field of UESR on the simultaneous removal of the heavy metals and organic contaminants were studied. The removal efficiencies of p-xylene and phenanthrene were higher in the experiments with cells of smaller diameter or larger height, and with distilled water flow in the cathode chamber. The removal efficiency of Cu and Pb were higher in the experiments with smaller diameter or shorter height cells and 0.01M HNO(3) solution as cathode chamber flow. In spite of different conditions for removal of heavy metals and organics, it is possible to use the upward electrokinetic soil remediation process for their simultaneous removal. Thus, in the experiments with duration of 6 days removal efficiencies of phenanthrene, p-xylene, Cu and Pb were 67%, 93%, 62% and 35%, respectively. The experiment demonstrated the feasibility of simultaneous removal of organic contaminants and heavy metals from kaolin using the upward electrokinetic soil remediation process. PMID- 17110024 TI - Seasonal trends in coarse and fine particle sources in Delhi by the chemical mass balance receptor model. AB - A study of the source contribution of atmospheric particulate matter and associated heavy metal concentrations using chemical mass balance model Version 8 (CMB8) in coarse and fine size mode has been carried out for the city of Delhi. Urban particles were collected using a five-stage impactor at six sites in three different seasons, viz. winter, summer and monsoon in the year 2001. Five samples from each site in each season were collected. The results obtained indicate the dominance of vehicular pollutants in fine size mode, whilst the contribution in coarse mode to some extent is site specific but largely due to vehicular pollution and, soil and crustal dust. Seasons also play an important role but in coarse size fraction only. PMID- 17110025 TI - An LNG release, transport, and fate model system for marine spills. AB - LNGMAP, a fully integrated, geographic information based modular system, has been developed to predict the fate and transport of marine spills of LNG. The model is organized as a discrete set of linked algorithms that represent the processes (time dependent release rate, spreading, transport on the water surface, evaporation from the water surface, transport and dispersion in the atmosphere, and, if ignited, burning and associated radiated heat fields) affecting LNG once it is released into the environment. A particle-based approach is employed in which discrete masses of LNG released from the source are modeled as individual masses of LNG or spillets. The model is designed to predict the gas mass balance as a function of time and to display the spatial and temporal evolution of the gas (and radiated energy field). LNGMAP has been validated by comparisons to predictions of models developed by ABS Consulting and Sandia for time dependent point releases from a draining tank, with and without burning. Simulations were in excellent agreement with those performed by ABS Consulting and consistent with Sandia's steady state results. To illustrate the model predictive capability for realistic emergency scenarios, simulations were performed for a tanker entering Block Island Sound. Three hypothetical cases were studied: the first assumes the vessel continues on course after the spill starts, the second that the vessel stops as soon as practical after the release begins (3 min), and the third that the vessel grounds at the closest site practical. The model shows that the areas of the surface pool and the incident thermal radiation field (with burning) are minimized and dispersed vapor cloud area (without burning) maximized if the vessel continues on course. For this case the surface pool area, with burning, is substantially smaller than for the without burning case because of the higher mass loss rate from the surface pool due to burning. Since the vessel speed substantially exceeds the spill spreading rate, both the thermal radiation fields and surface pool trail the vessel. The relative directions and speeds of the wind and vessel movement govern the orientation of the dispersed plume. If the vessel stops, the areas of the surface pool and incident radiation field (with burning) are maximized and the dispersed cloud area (without burning) minimized. The longer the delay in stopping the vessel, the smaller the peak values are for the pool area and the size of the thermal radiation field. Once the vessel stops, the spill pool is adjacent to the vessel and moving down current. The thermal radiation field is oriented similarly. These results may be particularly useful in contingency planning for underway vessels. PMID- 17110026 TI - Effects of Cr3+, Cr6+ and tannery sludge on C and N mineralization and microbial activity in semi-arid soils. AB - The aim of this study was to evaluate the effect of two Cr species (Cr(3+) and Cr(6+)) on N and C mineralization and dehydrogenase activity in semi-arid soils. The Cr species (250 mg kg(-1)soil) were either added alone or mixed with tannery sludge (0.0125 g g(-1)) to three soils: cultivated soils, and outside and under the canopy of mesquite trees were then incubated for 180 days at 25 degrees C. Sole Cr(6+) addition had a higher inhibition of CO(2) production rate in cultivated soil (58-73%) than in soils under the canopy and outside the canopy. Soil outside the canopy amended with Cr(6+) showed the highest inhibition of dehydrogenase activity (40-100%) followed by cultivated and under the canopy soils. However, Cr(6+) added alone increased the inhibition of nitrification in soil outside the canopy (68-84%, from 30 to 120 days), followed by under the canopy and cultivated soils. The addition of tannery sludge to Cr(6+) significantly reduce the CO(2) production rate and dehydrogenase activity in all three soils, and increased the inhibition of nitrification in the following order: outside the canopy, cultivated and under the canopy soils. The addition of Cr(3+) or Cr(3+) plus tannery sludge either stimulated or inhibited CO(2) production rate, dehydrogenate activity and ammonification in the three soils in no clearly defined order. Measurement of dehydrogenase activity was the best tool for assessing the harmful effect of Cr(6+) on soil microbial activity in semi arid soils exposed for an extended period. PMID- 17110027 TI - Removal of methyl tert-butyl ether (MTBE) with Nafion. AB - A solid organic polymer, Nafion, is tested for the removal of methyl tert-butyl ether (MTBE) in water. Nafion with perfluorosulfonic acid backbone and terminal sulfonic acid groups has a surface acidity similar to 100% sulfuric acid, and has been commonly used as a strong-acid catalyst in many organic reactions. Sorption and subsequent transformation of MTBE were observed in batch experiments. The transformation of MTBE by porous nanocomposite Nafion SAC-13 to tert-butyl alcohol (TBA), acetone, isobutene and probably methanol was found. Subsequent transformation of TBA to acetone was also observed. Results suggest that transformational pathways may include hydrolysis, dehydrogenation and oxidation. Dissolved oxygen is needed for the oxidation of isobutene to acetone. As Nafion is insoluble in water, chemically stable, and regenerable, its use in packed-bed reactors for MTBE removal looks promising. PMID- 17110028 TI - United states regulations for siting LNG terminals: problems and potential. AB - The regulations being applied to liquefied natural gas (LNG) import terminal siting in the United States are reviewed. There are no requirements for exclusion zones to protect the public from LNG spills onto water. Serious problems with current practices used to determine exclusion zones on the land-based part of the facility are identified. Many of the questions that are considered relate to the use of computational fluid dynamic (CFD) models, which appear to offer the best potential for realistic modeling to determine vapor cloud exclusion zones that result from LNG spills into impounded areas with or without dispersion in the presence of other obstacles to the wind flow. Failure to use CFD models, which are already approved by the regulation, and continued use of practices which have been demonstrated to be in error, raises important questions of credibility as well as denies the applicant full use of scientific tools that are available to optimize the design of such facilities so as to best provide for safety of the public. PMID- 17110029 TI - Metformin therapy in a transgenic mouse model of Huntington's disease. AB - Huntington's disease (HD) is a hereditary neurodegenerative disease that leads to striatal degeneration and a severe movement disorder. We used a transgenic mouse model of HD (the R6/2 line with approximately 150 glutamine repeats) to test a new therapy for this disease. We treated HD mice with metformin, a widely used anti-diabetes drug, in the drinking water (0, 2 or 5mg/ml) starting at 5 weeks of age. Metformin treatment significantly prolonged the survival time of male HD mice at the 2mg/ml dose (20.1% increase in lifespan) without affecting fasting blood glucose levels. This dose of metformin also decreased hind limb clasping time in 11-week-old mice. The higher dose did not prolong survival, and neither dose of metformin was effective in female HD mice. Collectively, our results suggest that metformin may be worth further investigation in additional HD models. PMID- 17110030 TI - Noradrenaline inhibits substantia gelatinosa neurons in mice trigeminal subnucleus caudalis via alpha(2) and beta adrenoceptors. AB - The actions of noradrenaline (NA) in the substantia gelatinosa (SG) are important for their antinociceptive effects. In order to identify the possible mechanisms underlying NA actions in the SG of trigeminal subnucleus caudalis (Vc), the direct membrane effects were examined by gramicidin-perforated patch clamp recording using brain slice preparation from immature mice brainstem. The majority (60/71, 85%) of neurons tested were hyperpolarized by NA application, and these hyperpolarizing effects were mimicked both by the alpha(2) adrenergic agonist, clonidine (18/28, 64%) and the beta adrenergic agonist, isoproterenol (9/24, 38%). NA-induced hyperpolarizing effect was also blocked by the alpha(2) adrenergic antagonist, yohimbine in five out of six neurons tested. However, a minority (5/71, 7%) of neurons tested were depolarized by NA, and these depolarizing effects were mimicked by the alpha(1) adrenergic agonist, phenylephrine (11/26, 42%). NA-induced hyperpolarizing effects were maintained in the presence of tetrodotoxin (TTX), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), d,l-2-amino-5-phosphonopentanoic acid (AP5), picrotoxin and strychnine, a Na(+) channel, ionotropic glutamate receptor, GABA(A) and glycine receptor antagonists, respectively, indicating that the effects of NA are direct on the postsynaptic SG neurons. These results indicate that alpha(2) and beta adrenoceptor mediate inhibition, and alpha(1) adrenoceptor mediates facilitation of orofacial nociceptive processing in mouse trigeminal brainstem SG neurons by postsynaptic actions. PMID- 17110031 TI - Gene therapy using SOD1 protects striatal neurons from experimental stroke. AB - Reactive oxygen species contribute to neuronal death following cerebral ischemia. Prior studies using transgenic animals have demonstrated the neuroprotective effect of the antioxidant, copper/zinc superoxide dismutase (SOD1). In this study, we investigated whether SOD1 overexpression using gene therapy techniques in non-transgenic animals would increase neuronal survival. A neurotropic, herpes simplex virus-1 (HSV-1) vector containing the SOD1 gene was injected into the striatum either before or after transient focal cerebral ischemia. Striatal neuron survival at 2 days was improved by 52% when vector was delivered 12-15 h prior to ischemia and by 53% when vector delivery was delayed 2 h following ischemia. These data add to the growing literature, which suggests that an antioxidant approach, perhaps by employing gene therapy techniques, may be beneficial in the treatment of stroke. PMID- 17110032 TI - Investigations on multimodal sensory integration: texture, taste, and ortho- and retronasal olfactory stimuli in concert. AB - Perceptual interactions between odour and oral texture were explored in a study in which a cream odour was presented ortho- or retronasally at well-defined moments whilst milk-like foods with different viscosities, produced by adding a thickener, were present in the mouth. Gaseous (odour) and liquid (texture) pulses were presented using a specially-developed computer-controlled system of air dilution olfactometry and pumps. Odour pulses, lasting 2 s, were presented either during a 3-s period in which a liquid filled the oral cavity, during a 3-s period in which the liquid was manipulated orally or during the swallowing of the liquid. Subjects rated the intensity of overall flavour, thickness and creaminess. Perceived flavour intensity was reduced with increasing viscosity of the liquid, irrespective of whether or not the odour was presented ortho- or retronasally. The odour stimulus increased the intensities of thickness and creaminess, but only when the odour was presented retronasally that is as if the odour would have originated from the liquid. Furthermore, this enhancement was most pronounced when odours coincided with swallowing, less pronounced when odours coincided with oral manipulation and absent when presented during mouth filling. The results suggest that cross-modal interactions are the rule rather than the exception, provided that multi-modal sensory integration has occurred. PMID- 17110033 TI - Decreased blood-brain barrier permeability to fluorescein in streptozotocin treated rats. AB - Investigations of the blood-brain barrier (BBB) in diabetes have yielded contradictory results. It is possible that diabetes differentially affects paracellular and transcellular permeabilities via modulation of tight junction and transport proteins, respectively. Fluorescein (FL), a marker for paracellular permeability, is a substrate for the transport proteins organic anion transporter (OAT)-3 and multidrug resistance protein (MRP)-2 at the BBB. Furthermore, MRP-2 mediated efflux of FL can be upregulated by glucose. In this study, streptozotocin-induced diabetes led to decreased brain distribution of FL measured by in situ brain perfusion, consistent with activation of an efflux transport system for FL at the BBB. This change was paralleled by increased protein expression of MRP-2, but not OAT-3, in cerebral microvessels. These data indicate that diabetes may lead to changes in efflux transporters at the BBB and have implications for delivery of therapeutics to the central nervous system. PMID- 17110034 TI - Antihyperalgesic effects of loperamide in a model of rat neuropathic pain are mediated by peripheral delta-opioid receptors. AB - The possible antihyperalgesic and antiallodynic activity of loperamide, an opioid agonist which does not readily penetrate the blood-brain barrier, were examined in the spinal nerve ligation model of experimental neuropathic pain. Intraperitoneal (i.p.) injection of loperamide effectively reversed thermal hyperalgesia. In contrast, loperamide had minimal effects on cold allodynia and no effects on mechanical allodynia. The antihyperalgesic action of loperamide against noxious heat was antagonized by naltrindole, a delta-opioid receptor selective antagonist, but not by pretreatment with beta-funaltrexamine, a mu opioid receptor selective antagonist, or administration of nor-binaltorphimine, a kappa-opioid receptor selective antagonist. Furthermore, i.p. injection of [d Ala(2), Glu(4)]-deltorphin II, a delta-opioid receptor selective peptide agonist, also reversed thermal hyperalgesia. The present results suggest that thermal hyperalgesia in experimental neuropathic pain can be reduced through activation of peripheral delta-opioid receptors. The data suggest the possible application of peripherally restricted and delta-opioid receptor selective agonists in the treatment of some aspects of neuropathic pain without many of the side effects associated with centrally acting opioids and without the peripheral side effects of opioid agonists acting at mu-receptors. PMID- 17110035 TI - Expression of Iba1 protein in microglial cells of zitter mutant rat. AB - Microglial activation has been associated with the pathogenesis of neurodegenerative disease. To characterize microglial responses in the zitter mutant rat, which shows progressive spongy degeneration, the development of microglial cells was investigated using ionized calcium-binding adaptor molecule (Iba1) antibody as a specific marker of microglial cells. Neurochemical analysis showed transiently increased Iba1 protein levels in the brains of developing Sprague-Dawley (SD) rats. However, high Iba1 protein readings continued in aged zitter rats. Immunohistochemical analysis revealed time-course differences in the transformation of microglia between SD and zitter rats and prolonged activation of microglial cells in the zitter rat. In the zitter rat, activated microglial cells characterized by swollen cell bodies and shorter, thicker processes were distributed throughout the brain from 2-weeks- to 2-months-old. After 2-months old, numbers of activated microglial cells gradually decreased. However, these cells were not observed in SD rats. Iba1-immunoreactive cell-clusters organized by at least five activated microglial cells were also prominent in the zitter brain. These differences reflect the neuropathology of this mutant rat triggered by deletion of the attractin gene. The present data may thus suggest that microglial cells directly or indirectly contribute to progressive spongy degeneration in zitter mutant rats. PMID- 17110036 TI - Expression of gonadotropin-releasing hormone receptor in cerebral cortical neurons of embryos and adult rats. AB - Mammalian gonadotropin-releasing hormone (GnRH) was initially isolated from hypothalamus and its receptor from anterior pituitary, although extrapituitary GnRH receptors have been reported. The aim of the present study was to investigate whether GnRH receptor and its mRNA are expressed in cerebral cortical neurons of rat embryos and adult rats using immunohistochemical and reverse transcriptase polymerase chain reaction (RT-PCR) techniques. The immunohistochemistry and RT-PCR analysis showed expression of GnRH receptor and presence of its mRNA, in both cerebral cortical neurons of rat embryos and cerebral cortical tissues of adult rats. Additional experiments showed a decrease in the receptor mRNA expression when cultured neurons of rat embryos were treated with GnRH. It is possible that the presence of GnRH receptors in cortical neurons of rat may be involved in other physiological roles such as neurohormone or neuromodulator. PMID- 17110037 TI - Postnatal development of septal projections to the midbrain central gray in female rats: tract-tracing analysis with DiI. AB - The neural projection of the lateral septum (LS) to the rostral mesencephalic central gray (MCG) is sexually dimorphic and plays an important role in inhibiting female reproductive behavior. In this experiment, development of the LS-MCG connection from birth to 15 days after birth was examined in female rats by a tract-tracing method with DiI. On the birth day (D1 rat), and 5, 10 or 15 days after birth (D5, D10 or D15 rat, respectively) or 8 weeks after birth (adult), the brain was fixed by perfusion of a mixture of 4% PFA and 0.1% glutaraldehyde. DiI was pasted on the coronally cut-surface of the LS and the sample was incubated in PFA at 40 degrees C for up to 4 months. After incubation, 200-microm frozen parasagittal sections were prepared and observed by fluorescence microscopy. As a result, numerous DiI labeled fibers were found in the preoptic area, the anterior and posterior hypothalamus, and the MCG in adult rats. In D1 rats, several labeled axons extended caudal to the anterior hypothalamic area. In D5 rats, a few labeled fibers reached the MCG. Some labeled fibers were observed in the rostral MCG of D10 rats. In D15 rats, a considerable number of labeled fibers were seen to reach the rostral MCG and relative density of the fibers was comparable to that of adult. These results suggest that the neural pathway from the LS to the rostral MCG develops acutely during the period from 5-10 days up to more than 15 days after birth. PMID- 17110038 TI - Cannabinoid CB1 receptor antagonists cause status epilepticus-like activity in the hippocampal neuronal culture model of acquired epilepsy. AB - Status epilepticus (SE) is a major medical emergency associated with a significant morbidity and mortality. Little is known about the mechanisms that terminate seizure activity and prevent the development of status epilepticus. Cannabinoids possess anticonvulsant properties and the endocannabinoid system has been implicated in regulating seizure duration and frequency. Endocannabinoids regulate synaptic transmission and dampen seizure activity via activation of the presynaptic cannabinoid receptor 1 (CB1). This study was initiated to evaluate the role of CB1 receptor-dependent endocannabinoid synaptic transmission towards preventing the development of status epilepticus-like activity in the well characterized hippocampal neuronal culture model of acquired epilepsy using patch clamp electrophysiology. Application of the CB1 receptor antagonists SR141716A (1 microM) or AM251 (1 microM) to "epileptic" neurons caused the development of continuous epileptiform activity, resembling electrographic status epilepticus. The induction of status epilepticus-like activity by CB1 receptor antagonists was reversible and could be overcome by maximal concentrations of CB1 agonists. Similar treatment of control neurons with CB1 receptor antagonists did not produce status epilepticus or hyperexcitability. These findings suggest that CB1 receptor-dependent endocannabinoid endogenous tone plays an important role in modulating seizure frequency and duration and preventing the development of status epilepticus-like activity in populations of epileptic neurons. The regulation of seizure activity and prevention of status epilepticus by the endocannabinoid system offers an important insight into understanding the basic mechanisms that control the development of continuous epileptiform discharges. PMID- 17110039 TI - Induction of total insensitivity to capsaicin and hypersensitivity to garlic extract in human by decreased expression of TRPV1. AB - TRPV1 is a cation channel which is activated by temperature (> or =42 degrees C) and capsaicin. In the present study, we found a person with total insensitivity to capsaicin and attempted to unravel its causes. The expression levels of TRPV1 protein and mRNA in the cells of the person's buccal mucosa were less than half of those in a normal subject. Sequential analysis of mRNA and genomic DNA revealed several point mutations mostly in the second intron of the person's TRPV1. Interestingly, the subject showed hypersensitivity to garlic extract, but TRPA1 (allicin receptor) level was normal. These results suggest that the decreased expression of TRPV1 may be related to a functional knock out in capsaicin sensation and hypersensitivity to allicin in humans. PMID- 17110040 TI - Spinal microglia and neuropathic pain in young rats. AB - Neuropathic pain behaviour is not observed in neonatal rats and tactile allodynia does not develop in the spared nerve injury (SNI) model until rats are 4 weeks of age at the time of surgery. Since activated spinal microglia are known to play a key role in neuropathic pain, we have investigated whether the microglial response to nerve injury in young rats differs from that in adults. Here we show that dorsal horn microglial activation, visualised with IBA-1 immunostaining, is significantly less in postnatal day (P) 10 rat pups than in adults, 7 days after SNI. This was confirmed by qPCR analysis of IBA-1 mRNA and mRNA of other microglial markers, integrin-alpha M, MHC-II DMalpha and MHC-II DMbeta. Dorsal horn IBA-1+ve microglia could be activated, however, by intraspinal injections of lipopolysaccharide (LPS) or N-methyl-d-aspartate (NMDA) at P10, although the increase in the levels of mRNA for all microglial markers was less than in the adult rat. In addition, P10 rats developed a small but significant mechanical allodynia in response to intrathecal LPS. Intrathecal injection of cultured ATP activated microglia, known to cause mechanical allodynia in adult rats, had no behavioural effect at P10 and only began to cause allodynia if injections were performed at P16. The results clearly demonstrate immaturity of the microglial response triggered by nerve injury in the first postnatal weeks which may explain the absence of tactile allodynia following peripheral nerve injury in young rats. PMID- 17110041 TI - Sarcoptic mange and metapodial development in growing male Iberian ibex (Capra pyrenaica). AB - The effect of mange (Sarcoptes scabiei) on metapodial growth was studied in 24 uninfested and 16 infested male Iberian ibex from Sierra Nevada National Park (southern Spain). Our results show that infested ibex had smaller metatarsal bones, lower body weight, and a delayed rhythm of ossification, by about 1 year. We conclude that skeletal development and body weight in the Iberian ibex are sensitive to sarcoptic mange. PMID- 17110042 TI - Characterization of circulating lymphocyte subpopulations in canine leishmaniasis throughout treatment with antimonials and allopurinol. AB - Canine leishmaniasis (CL) is a systemic parasitic disease with a wide variability of response to specific therapy: the majority of patients apparently improve with treatment, some of them respond but later relapse, and few of them do not respond at all. It has been demonstrated that the immune response plays a key role in the development and outcome of Leishmania infection in the dog and in the response to the treatment, although this response is not well understood. Some authors have suggested that ill dogs show a reduction in the percentage of circulating CD4+ lymphocytes and in the CD4+/CD8+ ratio, both of which normalize after treatment and clinical recovery. The present paper discusses the variation of the different lymphocyte subpopulations (CD3, CD4, CD8, CD21) of peripheral blood mononuclear cells (PBMC) in 28 dogs diagnosed with CL and submitted to conventional treatment with meglumine antimoniate (Glucantime) for 1 month and with allopurinol (Zyloric) for 1 year, in order to evaluate the usefulness of these parameters as indicators of the immunological condition of the ill animals and of the prognosis of their evolution during the treatment. It is concluded that circulating lymphocyte subpopulations are similar in dogs with leishmaniasis and in healthy dogs and that there is no correlation between the clinical status or response to therapy and the values of the counts of the different lymphocyte subpopulations. Therefore, the percentage of different lymphocyte subpopulations cannot be used as a parameter to predict the evolution of an individual patient in a clinical context. PMID- 17110043 TI - The first finding of a natural infection of Cryptosporidium muris in a cat. AB - Little is known about the species of Cryptosporidium infecting cats. The limited number of genetic studies conducted to date, have all identified C. felis as the species of Cryptosporidium in cats. We report a morphological and genetic description of a natural C. muris infection in a cat. Oocysts were viable and were successfully transmitted to laboratory mice. Further studies are required to determine the range and prevalence of Cryptosporidium species infecting cats. PMID- 17110044 TI - IpaD is localized at the tip of the Shigella flexneri type III secretion apparatus. AB - Type III secretion (T3S) systems are used by numerous Gram-negative pathogenic bacteria to inject virulence proteins into animal and plant host cells. The core of the T3S apparatus, known as the needle complex, is composed of a basal body transversing both bacterial membranes and a needle protruding above the bacterial surface. In Shigella flexneri, IpaD is required to inhibit the activity of the T3S apparatus prior to contact of bacteria with host and has been proposed to assist translocation of bacterial proteins into host cells. We investigated the localization of IpaD by electron microscopy analysis of cross-linked bacteria and mildly purified needle complexes. This analysis revealed the presence of a distinct density at the needle tip. A combination of single particle analysis, immuno-labeling and biochemical analysis, demonstrated that IpaD forms part of the structure at the needle tip. Anti-IpaD antibodies were shown to block entry of bacteria into epithelial cells. PMID- 17110045 TI - Engineering the sialic acid in organs of mice using N-propanoylmannosamine. AB - Sialic acids play an important role during development, regeneration and pathogenesis. The precursor of most physiological sialic acids, such as N acetylneuraminic acid is N-acetyl-D-mannosamine. Application of the novel N propanoylmannosamine leads to the incorporation of the new sialic acid N propanoylneuraminic acid into cell surface glycoconjugates. Here we analyzed the modified sialylation of several organs with N-propanoylneuraminic acid in mice. By using peracetylated N-propanoylmannosamine, we were able to replace in vivo between 1% (brain) and 68% (heart) of physiological sialic acids by N propanoylneuraminic acid. The possibility to modify cell surfaces with engineered sialic acids in vivo offers the opportunity to target therapeutic agents to sites of high sialic acid concentration in a variety of tumors. Furthermore, we demonstrated that application of N-propanoylmannosamine leads to a decrease in the polysialylation of the neural cell adhesion molecule in vivo, which is a marker of poor prognosis for some tumors with high metastatic potential. PMID- 17110046 TI - Oxidative stress and dopamine depletion in an intrastriatal 6-hydroxydopamine model of Parkinson's disease. AB - Although the etiology of Parkinson's disease (PD) is unknown, a common element of most theories is the involvement of oxidative stress, either as a cause or effect of the disease. There have been relatively few studies that have characterized oxidative stress in animal models of PD. In the present study a 6-hydroxydopamine (6-OHDA) rodent model of PD was used to investigate the in vivo production of oxidative stress after administration of the neurotoxin. 6-OHDA was injected into the striatum of young adult rats and the production of protein carbonyls and 4 hydroxynonenal (HNE) was measured at 1, 3, 7, and 14 days after administration. A significant increase in both markers was found in the striatum 1 day after neurotoxin administration, and this increase declined to basal levels by day 7. There was no significant increase found in the substantia nigra at any of the time points investigated. This same lesion paradigm produced dopamine depletions of 90-95% in the striatum and 63-80% in the substantia nigra by 14-28 days post-6 OHDA. Protein carbonyl and HNE levels were also measured in middle-aged and aged animals 1 day after striatal 6-OHDA. Both protein carbonyl and HNE levels were increased in the striatum of middle-aged and aged animals treated with 6-OHDA, but the increases were not as great as those observed in the young adult animals. Similar to the young animals, there were no increases in either marker in the substantia nigra of the middle-aged and aged animals. There was a trend for an age-dependent increase in basal amounts of oxidative stress markers when comparing the non-lesioned side of the brains of the three age groups. These results support that an early event in the course of dopamine depletion following intrastriatal 6-OHDA administration is the generation of oxidative stress. PMID- 17110047 TI - Localization of P2X2 and P2X3 receptors in rat trigeminal ganglion neurons. AB - Purine receptors have been implicated in central neurotransmission from nociceptive primary afferent neurons, and ATP-mediated currents in sensory neurons have been shown to be mediated by both P2X3 and P2X2/3 receptors. The aim of the present study was to quantitatively examine the distribution of P2X2 and P2X3 receptors in primary afferent cell bodies in the rat trigeminal ganglion, including those innervating the dura. In order to determine the classes of neurons that express these receptor subtypes, purine receptor immunoreactivity was examined for colocalization with markers of myelinated (neurofilament 200; NF200) or mostly unmyelinated, non-peptidergic fibers (Bandeiraea simplicifolia isolectin B4; IB4). Forty percent of P2X2 and 64% of P2X3 receptor-expressing cells were IB4 positive, and 33% of P2X2 and 31% of P2X3 receptor-expressing cells were NF200 positive. Approximately 40% of cells expressing P2X2 receptors also expressed P2X3 receptors and vice versa. Trigeminal ganglion neurons innervating the dura mater were retrogradely labeled and 52% of these neurons expressed either P2X2 or P2X3 or both receptors. These results are consistent with electrophysiological findings that P2X receptors exist on the central terminals of trigeminal afferent neurons, and provide evidence that afferents supplying the dura express both receptors. In addition, the data suggest specific differences exist in P2X receptor expression between the spinal and trigeminal nociceptive systems. PMID- 17110048 TI - Neurochemical characterization of pathways expressing plasma membrane monoamine transporter in the rat brain. AB - Neurotransmitter transporters play an important role in the control of synaptic transmission by ensuring the clearance of transmitters liberated in the synaptic cleft. In the case of monoaminergic neurotransmitters, this clearance is carried out by high-affinity reuptake transporters located in the plasma membrane of the presynaptic terminals. Recently plasma membrane monoamine transporter (PMAT), a transporter from the SLC29 (equilibrative nucleoside transporter) family, was shown to transport in vitro monoaminergic neurotransmitters, in particular dopamine and serotonin, nearly as efficiently as the high-affinity transporters. This transporter, well expressed in CNS, represents an interesting candidate for the control and modulation of aminergic pathways. We performed an extensive study of the distribution of PMAT in the rat brain. Our results highlight PMAT expression in brain regions which play a pivotal role in significant CNS functions and human neuropathologies. Using in situ hybridization immunohistochemistry co-labeling, PMAT mRNA was found in various neuron subtypes, including glutamatergic neurons of the hippocampus, mitral cells of the olfactory bulbs and GABAergic neurons in the substantia nigra pars reticulata and hypothalamus. Paradoxically, rat PMAT mRNA was found in some but not all monoaminergic nuclei. It was on the contrary predominantly expressed in major cholinergic groups throughout the brain, including brainstem motor nuclei, components of the basal forebrain cholinergic system and cholinergic interneurons of the striatum. These systems, implicated in locomotion, associative and spatial memory and reward-related learning, are disrupted at early stages of Parkinson's and Alzheimer's disease. Taken together, our observations support a role for PMAT in monoamine uptake in cholinergic neurons. PMID- 17110049 TI - T-type Ca2+ channels mediate propagation of odor-induced Ca2+ transients in rat olfactory receptor neurons. AB - Propagation of odor-induced Ca(2+) transients from the cilia/knob to the soma in mammalian olfactory receptor neurons (ORNs) is thought to be mediated exclusively by high-voltage-activated Ca(2+) channels. However, using confocal Ca(2+) imaging and immunocytochemistry we identified functional T-type Ca(2+) channels in rat ORNs. Here we show that T-type Ca(2+) channels in ORNs also mediate propagation of odor-induced Ca(2+) transients from the knob to the soma. In the presence of the selective inhibitor of T-type Ca(2+) channels mibefradil (10-15 microM) or Ni(2+) (100 microM), odor- and forskolin/3-isobutyl-1-methyl-xanthine (IBMX) induced Ca(2+) transients in the soma and dendrite were either strongly inhibited or abolished. The percentage of inhibition of the Ca(2+) transients in the knob, however, was 40-50% less than that in the soma. Ca(2+) transients induced by 30 mM K(+) were partially inhibited by mibefradil, but without a significant difference in the extent of inhibition between the knob and soma. Furthermore, an increase of as little as 2.5 mM in the extracellular K(+) concentration (7.5 mM K(+)) was found to induce Ca(2+) transients in ORNs, and such responses were completely inhibited by mibefradil or Ni(2+). Total replacement of extracellular Na(+) with N-methyl-d-glutamate inhibited none of the odor-, forskolin/IBMX- or 7.5 mM K(+)-induced Ca(2+) transients. Positive immunoreactivity to the Ca(v)3.1, Ca(v)3.2 and Ca(v)3.3 subunits of the T-type Ca(2+) channel was observed throughout the soma, dendrite and knob. These data suggest that involvement of T type Ca(2+) channels in the propagation of odor-induced Ca(2+) transients in ORNs may contribute to signal transduction and odor sensitivity. PMID- 17110050 TI - Sequence analysis and expression mapping of the rat clustered protocadherin gene repertoires. AB - Three closely-linked clusters of protocadherin (Pcdh) genes (alpha, beta, and gamma) encoding more than 50 distinct mRNAs have been identified in humans and mice, and proposed to play important roles in neuronal connectivity in the CNS. The human and mouse Pcdh alpha and gamma clusters each span a region of about 300 kb genomic DNA, and are each organized into a tandem array of more than a dozen highly-similar "variable" exons, and three downstream "constant" exons. Little is known about the expression patterns of the alpha and gamma repertoires in the CNS. Here, we comprehensively analyzed the one megabase rat Pcdh genomic DNA sequences at the nucleotide level using various computational methods. We found that the clustered rat Pcdh genes display strict orthologous relationships with those of mice but not humans. Moreover, each rat Pcdh variable exon is preceded by a distinct promoter. We designed two complete sets of isoform-specific probes and extensively mapped the expression patterns for each member of the alpha and gamma repertoires in the adult rat CNS by non-isotopic in situ hybridization experiments. We found that most alpha and gamma mRNA isoforms are broadly expressed in similar patterns in subsets of cells (with some displaying interesting cortical layer-specific expression) throughout various CNS regions, including the olfactory bulb, cerebral cortex, hippocampus, cerebellum, and spinal cord. The broad expression of most alpha or gamma mRNAs throughout various regions of the CNS is consistent with the hypothesis that these genes may be used for neurons to establish their individuality and also provide the adhesive diversity required for complex synaptic connectivity in the mammalian CNS. PMID- 17110052 TI - Negative cocaine effect expectancies are associated with subjective response to cocaine challenge in recreational cocaine users. AB - Although many studies have shown that cognitive effect expectancies are associated with drug use and drug treatment outcomes, few studies have compared effect expectancies with drug response following drug challenge. Healthy male and female volunteers (n=19, ages 21-35) who reported using cocaine 1-4 times per month completed the Cocaine Effect Expectancy Questionnaire (CEEQ: [Schafer, J. and Brown, S.A. (1991). Marijuana and cocaine effect expectancies and drug use patterns. Journal of Consulting and Clinical Psychology, 59, 558-565.]), were challenged with cocaine (0.9 mg/kg, i.n.), then completed a series of visual analog scales (VAS) and the Addiction Research Center Inventory (ARCI) at 15 min intervals for 3 h following cocaine administration. Significant positive correlations were found between global negative expectancies and peak responses on the VAS measures "Good," "Happy," "High," "Stimulated," and "Desire to Use Cocaine," and on the LSD subscale of the ARCI post-cocaine administration, and between global positive expectancies and the MBG subscale of the ARCI, and on VAS items "Anxious" and "Good" post-cocaine administration. Global positive expectancies also were positively correlated with peak systolic blood pressure, and global negative expectancies with peak heart rate after cocaine administration. These results suggest that negative and positive effect expectancies both play a complex role in the subjective experience of cocaine effects, and thus likely in the progression of non-use to recreational use, in the transition to abuse, and in individualized treatment strategies. PMID- 17110051 TI - Glucocorticoids exacerbate hypoxia-induced expression of the pro-apoptotic gene Bnip3 in the developing cortex. AB - Neonatal administration of the synthetic glucocorticoid, dexamethasone (DEX) retards brain growth, alters adult behaviors and induces cell death in the rat brain, thereby implicating glucocorticoids as developmentally neuroendangering compounds. Glucocorticoids also increase expression of pro-apoptotic Bcl-2 family members and exacerbate expression of hypoxic responsive genes. Bnip3 is a pro apoptotic Bcl-2 family member that is upregulated in response to hypoxia. In these studies, we investigated the interactions of glucocorticoid receptor and hypoxia in the regulation of Bnip3 mRNA in cortical neurons. Using quantitative real time reverse transcription-polymerase chain reaction, we found that DEX treatment of postnatal days 4-6 rat pups caused a significant increase in Bnip3 mRNA expression compared with vehicle controls. A significant increase in Bnip3 mRNA was also measured in primary cortical neurons 72 h after treatment with RU28362, a glucocorticoid receptor selective agonist. In primary cortical neurons, hypoxia increased Bnip3 mRNA expression and this was exacerbated with RU28362 treatment. To elucidate the mechanism of glucocorticoid- and hypoxia mediated regulation of Bnip3 transcription, a Bnip3 promoter-luciferase reporter construct was utilized in primary cortical neurons. Upregulation of the Bnip3 promoter was mediated by a single glucocorticoid response element and a hypoxic response element. Bnip3 overexpression in primary cortical neurons significantly increased cell death, which is dependent on the Bnip3 transmembrane domain. However, despite the increased expression of Bnip3 following glucocorticoid and hypoxia treatment, corresponding decreases in cell survival were minimal. These studies identify a novel pathway in the developing cortex through which glucocorticoids may enhance a metabolic insult, such as hypoxia. PMID- 17110053 TI - Occurrence of the acquired immunity in early vertebrates due to danger of transmissible cancers similar to canine venereal tumors. PMID- 17110054 TI - Mitogen-activated protein kinase (MAPK) intracellular signalling in the aqueous humour activated by Helicobacter pylori may have a role in glaucoma. PMID- 17110055 TI - Possible involvement of myofibroblast in the development of inflammatory aortic aneurysm. AB - To clarify the role of myofibroblasts in the development of inflammatory aortic aneurysm (IAA), 11 cases of IAA (69.2+/-8.59 years) were investigated immunohistochemically and were morphometrically compared with 12 age-matched cases of atherosclerotic abdominal aneurysm (AAA, 69.6+/-5.94 years). The positivity of mantle sign and CRP was significantly higher in the IAA group than in the AAA group. The wall of IAA (5.41+/-1.47 mm) was significantly thicker than that of AAA (2.68+/-0.71 mm). A significant increase in the expression of alpha smooth muscle actin was found in adventitial fibroblasts of IAA compared to those of AAA. The cell density and MIB-1 index of adventitial myofibroblasts were significantly higher in IAA than in AAA (cell density: 1.69+/-0.51 vs. 1.09+/-0.4 x 10(3) cells/mm(2); MIB-1 index: 5.25+/-2.97% vs. 1.55+/-0.71%). IAA showed a significantly lower area ratio (MAR) of adventitial microvessels than did AAA (2.92+/-1.49% vs. 7.51+/-2.64%). However, there was no significant difference in microvessel density (MVD) between IAA and AAA (84.62+/-50.5 vs. 65.1+/-32.6 vessels/mm(2)). In some cases of IAA, adventitial myofibroblasts expressed hypoxia-inducible factor 1alpha in their cytoplasm or nuclei while it was not detected in AAA. These findings suggest that the development of IAA may be partly mediated by the proliferation of adventitial myofibroblasts, which might be related to tissue hypoxia. PMID- 17110056 TI - Comparison of three interventions to increase mammography screening in low income African American women. AB - BACKGROUND: Low-income African American women are more likely to die of breast cancer than their Caucasian counterparts, and at least part of the difference in mortality results from differential screening adherence. The purpose of this study was to identify more efficacious methods of promoting routine mammography screening in underserved populations. METHODS: A prospective randomized intervention study of 344 low income African American women compared the impact of three interventions on mammography adherence and stage of readiness: (1) pamphlet only; (2) culturally appropriate video; and (3) interactive computer assisted instruction program. RESULTS: The interactive computer intervention program produced the greatest level of adherence to mammography (40.0%) compared to the video group (24.6%) and the pamphlet group (32.1%). When subjects in the pamphlet and video groups were combined to form a non-interactive group, this group had a significantly lower adherence than the group who received the interactive computer intervention (27.0% versus 40.0%). There was also significantly more forward movement in mammography stage of readiness among participants in the computer group (52.0%) compared to those in the pamphlet group (46.4%) or the video group (31.3%). When combining the non-interactive technology (pamphlet and video) there was also more forward movement in mammography stage of readiness for those in the interactive intervention group (52.0% moved 1 or 2 stages) compared to those in the non-interactive group (36.2%). CONCLUSIONS: These data indicate that tailored approaches are more effective than targeted messages either in print or video format. Another finding of this study is that interactive interventions are more effective than non interactive interventions in increasing adherence and moving African American women forward in their mammogram stage of readiness. PMID- 17110057 TI - Temporal discrimination learning by pigeons. AB - Memory for time by animals appears to undergo a systematic shortening. This so called choose-short effect can be seen in a conditional temporal discrimination when a delay is inserted between the sample and comparison stimuli. We have proposed that this temporal shortening may result from a procedural artifact in which the delay appears similar to the intertrial interval and thus, produces an inadvertent ambiguity or 'instructional failure'. When this ambiguity is avoided by distinguishing the intertrial interval from the delay, as well as the samples from the delay, the temporal shortening effect and other asymmetries often disappear. By avoiding artifacts that can lead to a misinterpretation of results, we may understand better how animals represent time. An alternative procedure for studying temporal discriminations is with the psychophysical bisection procedure in which following conditional discrimination training, intermediate durations are presented and the point of subjective equality is determined. Research using the bisection procedure has shown that pigeons represent temporal durations not only as their absolute value but also relative to durations from which they must be discriminated. Using this procedure, we have also found that time passes subjectively slower when animals are required to respond to the to-be-timed stimulus. PMID- 17110058 TI - Prevalence and correlates of opiate overdose among young injection drug users in a large U.S. city. AB - OBJECTIVES: The current study examines the prevalence and correlates of witnessing and experiencing opiate overdoses among a sample of young, injection drug users (IDUs) and non-injection drug users (NIDUs) in Baltimore, MD. METHODS: Data were derived from a longitudinal study of 15-30 year old IDUs and NIDUs (N=309) who had initiated heroin, cocaine, and/or crack use within 5 years prior to study enrollment. Chi-square and Wilcoxon rank-sum tests were used in bivariate analyses of demographic and drug use variables with each of the two dependent variables. Multivariate logistic regression models were used to identify correlates of experiencing and witnessing overdose. RESULTS: Twenty-nine percent of participants reported having ever experienced an opiate overdose and 57% reported having ever witnessed an overdose. Having ever experienced an opiate overdose was independently associated with being White (Adjusted Odds Ratio [AOR]=3.2; 95% Confidence Interval [CI]: 1.6, 6.4) recent homelessness (AOR=2.9; 95%CI: 1.5, 5.7); and length of injection, 5.6-6.9 years versus <5.6 years (AOR=4.0; 95%CI: 1.8-8.9); injecting 7.0-7.9 years versus <5.6 years (AOR=2.5; 95%CI: 1.03-6.1); injecting >8 versus <5.6 years (AOR=4.7; 95%CI: 2.2-10.2). Having witnessed an opiate overdose was independently associated with being White (AOR=2.4; 95%CI: 1.4, 4.1) and injecting >8 years versus <5.6 years (AOR=2.2; 95%CI: 1.2, 4.0). CONCLUSIONS: This study documents the high prevalence of witnessing and experiencing opiate overdoses among young, newly initiated IDUs and NIDUs. The results could inform the growing number of overdose prevention efforts throughout the U.S. PMID- 17110059 TI - Antimicrobial susceptibility patterns of Haemophilus parasuis from pigs in the United Kingdom and Spain. AB - A total of 30 British and 30 Spanish Haemophilus parasuis isolates were tested for their susceptibility to 19 of the antimicrobials currently used in swine practice with a broth microdilution method in order to know the emergence of resistance against these compounds in this porcine pathogen. All the British isolates were susceptible to penicillin, ceftiofur, erythromycin, tilmicosin, enrofloxacin, and florfenicol, and most of them were susceptible to the remaining antimicrobials (the highest resistance rate found was of 20% to neomycin). In contrast, all the Spanish isolates were susceptible exclusively to florfenicol, and high proportions of resistance were encountered for penicillin, ampicillin, oxytetracycline, erythromycin, tilmicosin, tiamulin and trimethoprim+sulphamethoxazole; in addition, a bimodal or multimodal distribution, or tailing of Spanish isolates over the MIC range was observed for clindamycin, sulphonamides and tylosine tartrate, suggesting the development of acquired resistance. In addition, several multiresistance patterns were found among the Spanish isolates, 23.3% of them being resistant to at least eight antimicrobials, the same rate as that encountered for those being susceptible to all antimicrobials tested. This study showed that in general British H. parasuis isolates are susceptible to antimicrobial agents routinely used for treatment of porcine respiratory diseases; however, the Spanish isolates need a more continuous surveillance of their susceptibility patterns. PMID- 17110060 TI - Foetal and adult human CYP3A isoforms in the bioactivation of organophosphorothionate insecticides. AB - In humans organophosphorothionate pesticides (OPT) prenatal exposure has been demonstrated. Since OPT-induced neurodevelopmental effects may be due to in situ bioactivation by foetal enzymes, the catalytic activity of the foetal CYP3A7 toward chlorpyrifos (CPF), parathion (PAR), malathion (MAL) and fenthion (FEN) has been assessed by using recombinant enzymes. A comparison with the adult isoforms CYP3A4 and CYP3A5 has been also carried out. CYP3A7 was able to produce significant levels of oxon or sulfoxide from the four OPTs in the range of tested concentrations (0.05-200 microM). When the efficiencies of CYP3A isoforms were compared, the ranking, expressed as CLi values, were: CPF=3A4>3A5>3A7; PAR=3A4>>3A7>>3A5; MAL=3A4>3A7>3A5; FEN (sulfoxide formation)=3A4>3A5>>3A7. The CYP3A5 efficiency appeared to be more dependent on the single insecticide than its related isozyme CYP3A4. Our results indicate that the levels of toxic metabolite formed in situ by CYP3A7 from CPF, MAL and PAR but not from FEN have the chance to inhibit acetylcholinesterase, following prenatal exposure to OPTs. However, due to the smaller weight of foetal liver, the contribution to total OPT biotransformation is relatively low. On the other hand, our results clearly indicate that at low CPF concentrations, the formation of the non-toxic metabolites is highly favoured in the foetus. PMID- 17110061 TI - A comparative analysis of metabolism and viability in porcine oocytes during in vitro maturation. AB - The importance of oocyte quality cannot be overstated, because it impacts all subsequent events during development of the embryo, the fetus and even the resulting offspring. Oocyte metabolism plays a critical role in supporting developmental competence via multiple mechanisms. It is beginning to be understood that metabolic pathways not only affect cytoplasmic maturation but may control nuclear maturation as well. A complete understanding of the precise roles that metabolism plays in determining oocyte quality is crucial for developing efficient in vitro maturation systems to support acquisition of oocyte competence. To date, this pursuit has not been entirely successful. Work in our laboratory on porcine oocyte metabolism has elucidated some of the intricate control mechanisms at work within the oocyte, not only for energy production, but also encompassing progression of nuclear maturation, mitochondrial activity and distribution, and oxidative and ionic stresses. We hypothesize that by utilizing oocyte metabolic data, we can develop more appropriate in vitro maturation systems that result in increased oocyte and embryo developmental competence. PMID- 17110062 TI - Lactose: a definitive guide to polymorph determination. AB - Lactose is a well-known molecule capable of forming a number of different polymorphs with varied chemical and physical properties. To date, no definitive guide for distinguishing between polymorphs using simple analytical techniques has been available. The information presented in this article aims to provide a conclusive guide for identifying the polymorphs of lactose and to successfully unravel years of contradictory research. Data have been collected on single phase polymorphs, prepared from an identical source, adopting the use of in situ and ex situ powder X-ray diffraction, CCD-Raman, FT-IR and (13)C-(1)H cross-polarisation magic angle spinning NMR (CP-MASNMR) spectroscopy, in order to provide simple methods to discriminate between the polymorphs. PMID- 17110063 TI - Nanoparticles--a historical perspective. AB - The historical development of nanoparticles starting with Paul Ehrlich and then first attempts by Ursula Scheffel and colleagues and the extensive work by the group of Professor Peter Speiser at the ETH Zurich in the late 1960s and early 1970s are described from a personal point of view. Special attention is given to the years between 1970 and the early 1980s. Further developments resulting from this work are also followed, and focus is placed on especially interesting improvements such as nanoparticles for the delivery of drugs across the blood brain barrier (BBB) and PEGylated nanoparticles with a prolonged blood circulation time. PMID- 17110064 TI - Estimation of 5-fluorouracil-loaded ethylene-vinyl acetate stent coating based on percolation thresholds. AB - The drug percolation thresholds of 5-fluorouracil-loaded ethylene-vinyl acetate stent coatings were estimated to characterize their drug release behavior and mechanical properties. The stent coatings were prepared using 5-fluorouracil (5 FU) as antitumor drug and ethylene-vinyl acetate (EVA) as matrix forming material in different ratios. In vitro release assays were carried out exposing only one side of coating to pH 6.5 PBS. Based on the release profiles, the drug percolation thresholds were estimated as 0.21 of total porosity (corresponding to ca. 32%, w/w of the drug), which is in approximately agreement with the atomic force microscopy (AFM) result. Based on the coating tensible break strength and tear break strength data, the mechanical percolation thresholds of drug were obtained as 39.7+/-0.3 and 37.5+/-1.4% (w/w) of drug content, respectively. PMID- 17110065 TI - Effect of traditional Chinese medicine Qin-Dan-Jiang-Ya-Tang on remodeled vascular phenotype and osteopontin in spontaneous hypertensive rats. AB - Qin-Dan-Jiang-Ya-Tang (QDJYT) is a traditional Chinese herbal medicine for the treatment of hypertension. The effect of QDJYT on blood pressure and on vascular remodeling in hypertension was investigated in the model of spontaneous hypertensive rats (SHR). Sixteen SHRs were divided into two groups, the SHR group and the SHR+QDJYT group. Eight WKY rats were a normal control group. QDJYT (750 mg/kg) was orally administered daily for 12 weeks in SHR+QDJYT group. After 12 weeks, thoracic aortas were segregated. Media thickness (MT), lumen diameter (LD), the ratio of MT to LD, the volume fraction of collagen (VFC) in media, the ultrastructure of vascular smooth muscle cells (VSMCs) and the expression of osteopontin (OPN) mRNA were examined by histological staining, transmission electron microscope (TEM), and real-time PCR, respectively. It was observed in our study that MT, MT/LD, VFC and the expression of OPN mRNA were higher in the SHRs than in the WKY rats, volume and numeral density of mitochondria in vascular smooth muscle cells (VSMCs) in media increased obviously. However, in the SHRs treated with QDJYT, we found MT, MT/LD, VFC and the expression of OPN gene were lower than in the SHRs, and the phenotype of VSMCs were close to normal. These results suggest that QDJYT could reverse the vascular remodeling in SHR, and the mechanisms may be related to the suppressive effect of QDJYT on the expression of OPN mRNA in arterial wall. PMID- 17110066 TI - Comparative pharmacokinetics of baicalin after oral administration of pure baicalin, Radix scutellariae extract and Huang-Lian-Jie-Du-Tang to rats. AB - Huang-Lian-Jie-Du-Tang (HLJDT) is an important "heat-clearing" multiherb remedy of traditional Chinese medicine, and Radix scutellariae (Scutellaria baicalensis Georgi, Labiatae) is a key ingredient herb in it. Baicalin and wogonoside are two main effective ingredients enriched in Radix scutellariae. In the present study, pharmacokinetic differences of baicalin following oral administration of pure baicalin, Radix scutellariae extract, baicalin co-administrated with extract of the other three herbs of HLJDT and HLJDT were investigated in male S.D. rats with approximately the same dose of 200 mg/kg baicalin. The pharmacokinetic comparison of wogonoside was conducted only in Radix scutellariae extract and HLJDT. Plasma concentrations of baicalin and wogonoside were determined using HPLC method. Unpaired Student's t-test was used for statistical comparison. The results indicated that baicalin and wogonoside demonstrated bimodal phenomenon in the plasma profile. Some ingredients in the other three herbs of HLJDT, not in Radix scutellariae itself, had pharmacokinetic interaction with baicalin and wogonoside and hence decreased their systematic exposure level (p<0.01). The absorption site of baicalin was preliminary evaluated in rat using in situ absorption in stomach and different intestinal segments and results revealed the existence of double site absorption of baicalin. The first absorption site was in upper intestinal, probably via directly absorption of baicalin; while the second absorption site was in colon in the form of aglygon. PMID- 17110067 TI - Antimicrobial and general toxicity activities of Gymnosperma glutinosum: a comparative study. AB - Gymnosperma glutinosum (Spreng.) Less (Asteraceae) is an important, and an effective herbal medicine which is wide used for the treatment of diarrhoea in Mexico. We examined and compared the antibacterial and antifungal activities through the dilution method and for general toxicity activity by the brine shrimp lethality assay of two samples of Gymnosperma glutinosum from two localities of Mexico: San Rafael-Coxcatlan (Puebla State) and Tepeji del Rio (Hidalgo State). In addition, two bioactive compounds (-)-17-hydroxy-neo-clerod-3-en-15-oic acid (1) and 5,7-dihydroxy-3,6,8,2',4',5'-hexamethoxyflavone (2) were isolated. From the hexane extract from both places was obtained a MeOH partition M(2). M(2) of Tepeji del Rio showed the least MICs (<125 microg/ml) in the majority of the bacterial strains. Sarcina lutea was the most sensitive bacteria (MIC< 125 microg/ml). The hexane extract of both localities showed antifungal activity against all tested fungi. San Rafael's hexane extract was significant more activity than Tepeji del Rio. Aspergillus niger (IC(50)=23.79 microg/ml) and Trichophyton mentagrophytes (IC(50)=90.25 microg/ml) were the more sensitive fungus strains. The strongest general toxicity activity was observed with the M(2) partition from Tepeji del Rio (LC(50)=503.7 microg/ml). The results obtained in this investigation, showed differences between the antimicrobial activities of the samples of plants collected in San Rafael (Puebla) and Tepeji del Rio (Hidalgo). PMID- 17110068 TI - Identification and analysis of human remains recovered from wells from the 1991 War in Croatia. AB - From 1996 to the present, the remains of 61 individuals killed during the 1991 War in Croatia were recovered from both dried out and functioning wells. Positive identification was established in 60.7% or 37/61 cases. Remains recovered from the same geographical region but from non-well settings were identified in 77.4% or 1256/1623 cases. The purpose of this paper is to report on the taphonomic, demographic and trauma characteristics of remains recovered from wells and identify factors responsible for the discrepancy in the identification ratios. The age and sex distributions in the well and non-well series were similar, as were the frequencies of recovered personal documents, jewelry and other artifacts. The taphonomic features of the remains were, however, significantly different. Preservation of remains was considerably better in non-well settings (1400/1623 or 86.3% compared to 40/61 or 65.5% in wells). Congruently, commingling of remains was more frequent in wells (26/61 or 42.6% compared to 77/1623 or 4.7% in non-well settings). In bodies recovered from non-well settings the preservation, state and commingling of the remains were strongly correlated with positive identification. None of these features were correlated with the identification of bodies from wells. Instead, identification of remains from wells was significantly affected by the presence or absence of water in the well. As both series have similar frequencies, and identical rankings of identification factors, the reason for the discrepancy in the identification ratios lies in the fact that by themselves, these factors were rarely sufficient for positive identification. In both series the majority of identifications (51.4% in the well, and 58.1% in the non-well series) were established through a combination of biological and non-biological features. The significantly lower identification ratio in the well series resulted from the difficult recovery conditions in wells with significant amounts of water, and the negative effect that water had on the preservation of clothing, personal artifacts and some dental features. Significant differences were also noted in the types and locations of perimortem traumas. There is a significantly higher frequency of entrance gunshot wounds to the back of the head, suggestive of execution style murders, in the well series. PMID- 17110069 TI - Fatal rupture of an undiagnosed aneurysm of the splenic artery--medico-legal implications. AB - Aneurysms of the visceral arteries are a rather common feature appearing in 0.1 2% of the population. The clinical relevance of those anomalies varies a lot. The wide range of descriptions reaches from asymptomatic cases to fatalities in particular due to haemorrhages. The latter will be discussed in a case report concerning a 60-year-old man who collapsed at his work place and died 4 h after admission to the intensive care unit of a hospital nearby. The ruptured aneurysm remained undiagnosed in hospital and was found by autopsy. The case is presented and discussed in view of medico-legal questions. PMID- 17110070 TI - Longevity and the stress response in Drosophila. AB - The concept that lifespan is a function of the capacity to withstand extrinsic stress is very old. In concordance with this, long-lived individuals often have increased resistance against a variety of stresses throughout life. Genes underlying the stress response may therefore have the ability to affect lifespan. The progress in modern genetic techniques has allowed researchers to test this idea. The general stress response involves the expression of stress proteins, such as chaperones and antioxidative proteins, downregulation of genes involved in energy metabolism and the release of protective substances. Do these same changes in patterns of expression have the ability to mitigate ageing and prolong lifespan? It appears that parts of this response indeed are also associated with extended longevity, whereas some elements are not, due to their high cost or long term deleterious consequences. Here we briefly review the state of the art of research on ageing and longevity in the model organism Drosophila, with focus on the role of the general stress response. We will conclude by contemplating some of the implications of the findings in this research and will suggest several directions for future research. PMID- 17110071 TI - Advances in endocrinology of aging research, 2005-2006. AB - The purpose of this brief review is to highlight some of the more important advances in endocrinology of aging research over the past year. Four advances were chosen and briefly described. First, exploration of the early steps in the generation of the internal steroidal hormonal signal involved in lifespan extension via the insulin/IGF-like signaling pathway in the nematode by two research groups revealed that the product of cholestanoic acid derivatives metabolized by a cytochrome P-450-like protein activates a protein with homology to the mammalian nuclear receptor superfamily, a process strikingly similar to the steroid hormone signaling pathway documented in mammalian systems. Second is the discovery that sirtuins, proteins that regulate lifespan in model organisms, enhance pancreatic insulin secretion in mice following a glucose challenge, suggesting the potential to regulate mammalian lifespan through regulation of the insulin signaling pathway. Third, the newly discovered hormone klotho, which also plays a role in regulating lifespan, in this case in mice, is reported to not only negatively affect insulin sensitivity but, perhaps more importantly, significantly affects calcium and phosphate metabolism as a required cofactor of Fgf-23 signaling. Finally the gonadotropin FSH is shown to directly affect bone density in mice separate from any direct effect of estrogen, suggesting that reproductive hormones other than estrogen can directly impact menopause associated pathophysiology in non-reproductive tissues. PMID- 17110072 TI - Radiological and clinical course of pneumonia in patients with avian influenza H5N1. AB - INTRODUCTION: We evaluated chest X-ray and clinical findings of patients with lower respiratory tract infection due to influenza H5N1 and presented the radiological findings and clinical course of the infection. MATERIALS AND METHODS: Between December 2005 and February 2006, eight hospitalized patients (median age 10, 5-15 years) with avian-flu were evaluated in this study. All patients were evaluated with chest X-ray and four of them with CT scan. Post mortem pathological characterization were also available for three of the patients. RESULTS: A rapidly progressive pneumonia with high mortality rate was observed especially for cases with late admission. The major radiologic abnormalities were extensive pneumonic infiltration with segmental and multifocal distribution, mostly located in lower zones of the lung. No pleural effusion and hilar lymphadenopathy was noted. CONCLUSION: Avian flu may be presented as rapidly progressive pneumonia. The chest radiography has an important role in diagnosis and should be obtained daily because of rapid change of the findings that may necessitate prompt action. PMID- 17110073 TI - Bioavailability and urinary excretion of isoflavones in humans: effects of soy based supplements formulation and equol production. AB - Soy isoflavones (IF) are of particular interest for their possible estrogenic effects on the symptoms of menopause. The bioavailability of IF is clearly a factor influencing their biological activity. The first aim of this study was to elucidate the impact of the matrix process and especially the formulation of soy based capsules on IF bioavailability. Twelve healthy volunteers were recruited for a randomized, double-blind, two-way crossover trial and received a single dose of the two soy-based formulations, one containing a pure soy standardized extract of IF, and the other containing soy flour in addition to the standardized extract of IF. Using a new and validated ELISA method, we measured the plasma and urinary concentrations of genistein, daidzein and its metabolite equol. Based on European Medicine Evaluation Agency recommendations, the main pharmacokinetic parameters allowed us to demonstrate the bioequivalence of the two formulations, indicating that the presence or absence of soy flour did not alter either the absorption or the elimination of daidzein and genistein. As bioequivalence was demonstrated, we pooled data collected during the two study-periods to address another original issue: Did the ability to produce equol affect the bioavailability of daidzein? We demonstrated that daidzein excretion was significantly lower in equol producers compared with equol non producers over the entire elimination period of the soy IF. This difference disappeared when equol excretion was added to daidzein excretion in equol producers. Our results indicated that the production of equol could partly explain the difference in daidzein bioavailability after IF ingestion. PMID- 17110074 TI - Endoscope-assisted retrosigmoid intradural suprameatal approach to the middle fossa: anatomical and surgical considerations. AB - INTRODUCTION: Lesions involving the posterior and middle fossa may be reached by several surgical approaches depending mostly on tumors' location and characteristics. The retrosigmoid intradural suprameatal approach has been used to remove tumors of the posterior fossa extending into Meckel's cave and the middle fossa. With large tumors, this approach may allow exposure of the oculomotor and optic nerves, the supraclinoid internal carotid and communicating posterior arteries. METHODS: Three formaldehyde-fixed specimens have been prepared on both sides using the conventional retrosigmoidal-suprameatal approach. The entire endoscopic preparations have been documented via a digital recording system, using a 5mm endoscope with a 25 degrees perspective. The authors describe the anatomical corridor and technique of the endoscope-assisted retrosigmoid intradural suprameatal approach to lesions that are located predominantly within the posterior fossa and supratentorial extension into Meckel's cave, sellar and parasellar region. CONCLUSIONS: The endoscope-assisted suprameatal approach provides exposure of the antero-medial middle fossa even in cases of skull base lesions that have not caused significant displacement of neurovascular structures. PMID- 17110075 TI - Outcomes of laparoscopic Miles' operation in very low rectal adenocarcinoma. Analysis of 32 cases. AB - AIMS: Minivasive techniques for excision of low rectal tumours have spread worldwide with good results, but their employment is still under discussion. The purpose of this study is to assess short term results and survival of laparoscopic abdominoperineal resection (LAPR) in very low rectal cancers. METHODS: The charts of 32 patients undergoing LAPR for very low rectal adenocarcinoma (0-2cm from dentata line) were reviewed retrospectively. Outcomes were evaluated considering surgical procedure, short and long-term results and survival. RESULTS: A thorough LAPR was performed in 31 patients and conversion to laparotomy was required in 1 patient. Mean operating time was 244min. The length of hospital stay (LOS) was 13,3days. The mean number of nodes collected was 12 and the distal margin was 3,6cm on average. There was 1 post-operative death. In the follow up no pelvic recurrence was observed, while metachronous metastases were observed in 5 patients and peritoneal carcinosis in 2 patients. No port site metastasis was registered. Cumulative 5year survival probability was 0,50. CONCLUSIONS: The outcomes of this study suggest that LAPR in very low rectal cancer is a reliable procedure, operating time and LOS were acceptable. Oncologic principles were respected: length of specimen, distal margin and number of nodes retrieved were quite acceptable. Pelvic recurrence frequency was nil. Long term results were comparable with those of other series. PMID- 17110076 TI - Doughnut mastopexy lumpectomy versus standard lumpectomy in breast cancer surgery: a prospective study. AB - AIMS: Doughnut mastopexy lumpectomy (DML) is a unique breast resection in which a tissue segment is removed and the breast reshaped through a periareolar incision. The present prospective investigation compares DML and standard lumpectomy (SL) in relation to surgical variables, histological parameters, postoperative morbidity and cosmetic outcome. METHODS: 127 patients with breast cancer were offered the choice between two conservative surgical approaches: doughnut mastopexy lumpectomy (DML group, n=39) or standard lumpectomy (SL group, n=88). The groups were comparable for radiological tumour size, tumour location within the breast, histological size, and pT category. Comparison was performed in term of surgical variables, histological parameters, postoperative morbidity and cosmetic outcome. RESULTS: The patients undergoing DML were younger than the patients who chose SL. In the DML group, the skin incision was 3-fold longer than in the SL group but was obtained with a final scar located around the nipple areola complex without further postoperative complications. The average volume of the breast specimen was higher in the DML group compared with SL group. The clinician assessment of cosmetic outcome reported a higher rate of acceptable result in the DML group than in the SL group. However, patient's assessment did not show difference of cosmetic satisfaction between groups. CONCLUSION: Our comparative study indicates that DML may be a useful alternative to SL not only in terms of accurate breast tissue resection but also in term of cosmetic results. PMID- 17110077 TI - Goals and intentions mediate efficacy beliefs and declining physical activity in high school girls. AB - BACKGROUND: According to theory, girls who set goals about increasing their physical activity and who are dissatisfied with their current activity level are likely to form intentions to be active and to carry out those intentions, especially if they have high efficacy and control beliefs about being physically active. We tested those ideas while observing naturally occurring change during high school. METHODS: A cohort of 431 black and white girls was tested at the end of their 9th- and 12th-grade academic years. Confirmatory factor analysis established the structural invariance of the measures across the 3-year study period. Structural equation modeling and panel analysis were used to determine whether changes in goal setting and satisfaction would mediate relations of self efficacy and perceived behavioral control with changes in intention and physical activity. Testing occurred between February and May in 1999 and 2004. Data were analyzed in 2006. RESULTS: Goal setting and intention mediated the indirect relation between self-efficacy and change in physical activity. Perceived behavioral control and physical activity change were related directly and also indirectly by a path mediated through satisfaction and intention. Black girls had lower self-efficacy, but changes in other variables were unrelated to race. CONCLUSIONS: These observations of longitudinal relations elaborate application of self-efficacy theory and the theory of planned behavior to physical activity by showing that goal setting and satisfaction mediate the relations of self efficacy and perceived behavioral control with changes in intention and physical activity. The results encourage additional research to identify the sources and development of physical activity goals, and their attainment, among girls, and whether experimental manipulation of goals and intentions can mitigate the decline in girls' physical activity during high school. PMID- 17110078 TI - Murine hepatoma (Hepa1c1c7) cells: a responsive in vitro system for chemoprotective enzyme induction by organoselenium compounds. AB - Murine (Hepa1c1c7) hepatoma cells are a suitable in vitro system for investigating the regulation of chemoprotective enzymes by selenazolidines, novel l-selenocysteine prodrugs developed as potential chemopreventive agents. They are less sensitive to the cytotoxic effects of both selenite and the less toxic selenazolidines than rat hepatoma (H4IIE) cells. All four selenazolidine 4 carboxylic acid (SCA) derivatives examined elevated thioredoxin reductase (Txnrd1), alpha-class glutathione transferases (Gsta), and UDP glucuronosyltransferase (Ugt)1a6 mRNAs. NAD(P)H-quinone oxidoreductase (Nqo1) was induced by the three 2-alkyl derivatives (2-cyclohexylSCA, 2-butylSCA, and 2 methylSCA) but not SCA itself. Transcripts of mu- and pi-class glutathione transferases were induced only by 2-cyclohexylSCA and 2-butylSCA. Only Gsta and Txnrd1 transcripts were elevated by l-selenomethionine, l-selenocystine, or Se methyl-l-selenocysteine. Txnrd1, Gsta, Nqo1, and Gstp responses to selenazolidines were all abolished by actinomycin D while Ugt1a6 responses were not. Induction responses to the selenazolidines were also eliminated (most) or reduced (Txnrd1 by 2-methylSCA) by cycloheximide, with the exception of Ugt1a6. The Ugt1a6 mRNA levels in the presence of SCAs and cycloheximide were similar to those with cycloheximide alone, and were almost double those of vehicle-treated cells. Thus, Hepa1c1c7 cells appear to provide a viable platform for the study of protective enzyme regulation by selenocompounds, and with the exception of Ugt1a6, the mRNA elevations from selenazolidines are transcriptionally dependent. PMID- 17110079 TI - The crude venom from the sea anemone Stichodactyla helianthus induces haemolysis and slight peroxidative damage in rat and human erythrocytes. AB - The haemolytic and peroxidative effects of crude venom of the sea anemone Stichodactyla helianthus were evaluated in rat and human erythrocytes. Venom extract caused a significant concentration-dependent effect on haemolysis (release of haemoglobin). Human erythrocytes were more sensitive (0.094 mg protein/ml) than those of the rats (0.3787 mg protein/ml). In contrast, a light effect on lipid peroxidation (LP, an index of oxidative damage to membrane lipids) was recorded. The concentrations needed to produce a significant effect on LP in rat and human erythrocytes were, respectively, 2-fold and 7-fold higher than those required to produce significant haemolysis. The differential effect of S. helianthus venom on haemolysis and oxidation of membrane lipids is not common for venoms of other sea anemones, which usually show a tightly related effect on LP and haemolytic damage. PMID- 17110080 TI - A self-report version of the Yale-Brown Obsessive-Compulsive Scale Symptom Checklist: psychometric properties of factor-based scales in three samples. AB - Current research in Obsessive-Compulsive Disorder (OCD) recognizes substantial symptom heterogeneity and emphasizes dimensional assessment of core domains. This study administered a self-report version of the Yale-Brown Obsessive-Compulsive Scale Symptom Checklist to OCD patients (n=53), non-OCD psychiatric patients (n=96), and students (n=419). Factor analyses of category- versus item-level data produced different solutions (4 or 5 vs. 3 factors, respectively), but support a multidimensional framework for OCD symptoms. For between-groups analyses, the two patient groups scored significantly higher than students on nearly all dimensions. However, OCD and non-OCD patients differed significantly only on Symmetry/Ordering symptoms. These findings provide novel data concerning this instrument and suggest that most of its scales may not distinguish OCD patients clearly when administered in this manner. We provide recommendations for improving subsequent self-report versions but caution users not to over-extend its intended use. PMID- 17110081 TI - Convergent validity of the aberrant behavior checklist and behavior problems inventory with people with complex needs. AB - The current study aimed to replicate and extend Rojahn et al. [Rojahn, J., Aman, M. G., Matson, J. L., & Mayville, E. (2003). The aberrant behavior checklist and the behavior problems inventory: Convergent and divergent validity. Research in Developmental Disabilities, 24, 391-404] by examining the convergent validity of the behavior problems inventory (BPI) and the aberrant behavior checklist (ABC) for individuals presenting with multiple complex behavior problems. Data were collected from 69 children and adults with severe intellectual disabilities and challenging behavior living in residential establishments. MANCOVA analyses showed that individuals with elevated BPI stereotyped behavior subscale scores had higher scores on ABC lethargy and stereotypy subscales, while those with elevated BPI aggressive/destructive behavior subscale scores obtained higher scores on ABC irritability, stereotypy and hyperactivity subscales. Multiple regression analyses showed a corresponding pattern of results in the prediction of ABC subscale scores by BPI subscale scores. Exploratory factor analysis of the BPI data suggested a six-factor solution with an aggressive/destructive behavior factor, four factors relating to stereotypy, and one related to stereotypy and self-injury. These results, discussed with reference to Rojahn et al. [Rojahn, J., Aman, M. G., Matson, J. L., & Mayville, E. (2003). The aberrant behavior checklist and the behavior problems inventory: Convergent and divergent validity. Research in Developmental Disabilities, 24, 391-404], support the existence of relationships between specific subscales of the two instruments in addition to an overall association between total scores related to general severity of behavioral disturbance. PMID- 17110082 TI - The NPM-ALK tyrosine kinase mimics TCR signalling pathways, inducing NFAT and AP 1 by RAS-dependent mechanisms. AB - Nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) expression is associated with the lymphoid malignancy anaplastic large cell lymphoma (ALCL) and results from a t(2;5) chromosomal translocation. We show that NPM-ALK induces Ras activation and phosphorylation of the ERK MAP Kinase consistent with activation of the Ras-MAP Kinase pathway. Furthermore, we demonstrate that activation of Ras is necessary for inducing transcription via NFAT/AP-1 composite transcriptional binding sites. This activity is dependent on NPM-ALK forming complexes with proteins that bind to autophosphorylated tyrosine residues at positions 156, 567 and 664, associated with binding to IRS-1, Shc and PLCgamma, respectively. Specifically, NPM-ALK activates transcription from the TRE promoter element, an AP-1 binding region, an activity dependent on both Ras and Shc activity. Our results show that NPM-ALK mimics activated T-cell receptor signalling by inducing pathways associated with the activation of NFAT/AP-1 transcription factors that bind to promoter elements found in a broad array of cytokine genes. PMID- 17110083 TI - Aspiration of an impacted lower third molar during its surgical removal under local anaesthesia. AB - In this case of an aspirated impacted lower third molar during its removal under local anaesthesia, the problem was recognized immediately during the surgical procedure. The patient, a 23-year-old male, was subjected to urgent radiological examination. The aspirated tooth was detected in the right bronchus and eventually removed by rigid bronchoscopy. Oral surgeons should suspect any tooth that has been avulsed or extracted and not found as having been aspirated. Early diagnosis and management of such incidents is essential. PMID- 17110084 TI - Human papillomaviruses in oral squamous cell carcinoma and pre-cancerous lesions detected by PCR-based gene-chip array. AB - Human papillomavirus (HPV) infection is a significant risk factor for uterine cervical carcinoma. Many studies have also demonstrated the presence of HPV in oral epithelia tissue, but the role of HPV infection in oral squamous cell carcinoma (OSCC) is still controversial. The aim of this study was to determine the frequency and type of HPV in OSCC and oral pre-cancerous lesions. DNA samples were collected by cytobrushing from 51 patients with OSCC, 46 with oral pre cancerous lesions and 90 normal controls. Nested polymerase chain reaction and gene-chip arrays were used to identify the HPV types in the samples. In pre cancerous lesions, there was a higher frequency of HPV of any type (14/46, OR = 2.844, CI = 1.186-6.816, P = 0.0216) and of low-risk HPV types (9/46, OR = 5.529, CI = 1.597-19.14, P = 0.0096) than in control samples. The prevalence of high risk types was significantly higher in OSCC than in control lesions (11/51 vs 8/90, OR = 2.819, CI = 1.051-7.558, P = 0.0420) but this was not the case for HPV of any type (13/51 vs 12/90, OR = 2.244, CI = 0.9266-5.337, P = 0.1066). High risk HPV types are prevalent in OSCC and may play a role in its progression, while low-risk types are associated with oral pre-cancerous lesions. PMID- 17110085 TI - Evaluation of expression of midkine in oral squamous cell carcinoma and its correlation with tumour angiogenesis. AB - Midkine, a new member of the heparin-binding growth factor family, was found recently to have a high expression level in many carcinoma specimens, including oesophagus, gall, bladder, pancreas, colorectum, breast and lung carcinomas, but few relevant studies have been performed. The aim of this study was to investigate the expression of midkine in oral squamous cell carcinoma (OSCC) and its correlation with tumour angiogenesis. Surgical specimens from 62 cases of OSCC and 10 cases of normal oral mucosa were examined by immunohistochemistry. The relationship between expression of midkine and the clinicopathological data and tumour angiogenesis was analysed. Midkine proteins were overexpressed in OSCC. The level of midkine expression was found to be significantly correlated with tumour size (P = 0.01), clinical stage (P = 0.002) and prognosis (P = 0.038). High midkine expression was associated with higher tumour angiogenesis as reflected by increased vascular endothelial growth factor expression (P = 0.005) and higher microvessel density (P = 0.004). These results indicate that midkine may be of great value in assessing the development and prognosis of OSCC, and could be taken as a new target for OSCC therapy. PMID- 17110086 TI - Corticotomy and compression osteogenesis in the posterior maxilla for treating severe anterior open bite. AB - A new technique is described for outpatient treatment of anterior open bite. The compression osteogenesis method with a two-stage corticotomy was used in the posterior maxilla to treat a woman with severe anterior open bite. Three-week post-surgical compression using anchor plates and elastics repositioned the posterior maxillary bone/teeth segments by 7 mm to the ideal superior position. The patient had a stable skeletal position of the maxilla at 14-month follow-up with satisfactory results and no complications after orthodontic treatment. This technique appears to be an efficient option for treating patients with anterior open bite. PMID- 17110087 TI - The burden of depressive symptoms in the long-term treatment of patients with schizophrenia. AB - OBJECTIVE: To prospectively measure the link between depressive symptoms and functional outcomes in the long-term treatment of people with schizophrenia. METHODS: Data were drawn from a large, multi-site, 3-year, prospective, naturalistic, observational study, in which subjects with schizophrenia were assessed at enrollment and at 12-month intervals thereafter. Individuals who were "Depressed" (defined as a total score > or =16 on the Montgomery-Asberg Depression Rating Scale) at enrollment were compared to those "Non-depressed" on functional outcomes, using self-report measures, clinicians' ratings, and information from medical records. Statistical analyses included Generalized Estimation Equation and mixed regression analyses adjusted for individual characteristics. Longitudinal group comparisons across the 3-year study were augmented with a cross-sectional group comparison at enrollment. RESULTS: At enrollment, 39.4% (877/2228) of the participants were deemed Depressed. Across the 3-year study, the depressed cohort was significantly more likely than the Non depressed to use relapse-related mental health services (emergency psychiatric services, sessions with psychiatrists); to be a safety concern (violent, arrested, victimized, suicidal); to have greater substance-related problems; and to report poorer life satisfaction, quality of life, mental functioning, family relationships, and medication adherence. Furthermore, changes in depressed status were associated with changes in functional outcomes. CONCLUSIONS: People with schizophrenia and concurrent depressive symptoms have poorer long-term functional outcomes compared to the Non-depressed. Their poorer quality of life, greater use of mental health services, and higher risk of involvement with law enforcement agencies underscore a need for special treatment interventions. Treatment of the non-psychotic dimensions of schizophrenia is a critical part of recovery. PMID- 17110088 TI - Epigenetic changes in estrogen receptor beta gene in atherosclerotic cardiovascular tissues and in-vitro vascular senescence. AB - Epigenetic changes marked by DNA methylation have been proposed to play a role in age-related disease. We investigated DNA methylation changes in cardiovascular atherosclerotic tissues and in-vitro vascular senescence in the promoter of estrogen receptor beta gene, which has essential roles in vascular function. Coronary atherosclerotic tissues showed higher methylation levels (28.7%) than normal appearing arterial (6.7%-10.1%) and venous tissues (18.2%). In comparing estrogen receptor beta methylation between plaque and non-plaque regions in ascending aorta, common carotid artery, and femoral artery of two patients, the plaque lesions showed consistently higher methylation levels than non-plaque regions. Passage-dependent increased estrogen receptor beta methylation was observed in three of six human aortic endothelial or smooth muscle cell lines cultured in-vitro to vascular senescence. Estrogen receptor beta expression in these vascular cell lines was significantly activated by DNA-methyltransferase inhibition. This activity was augmented by histone deacetylase inhibition. These findings provide evidence of epigenetic dysregulation of estrogen receptor beta in atherosclerosis and vascular aging. We suggest that focal epigenetic changes in estrogen receptor beta contribute to the development of atherosclerosis and vascular aging. PMID- 17110089 TI - Oculopharyngeal muscular dystrophy: recent advances in the understanding of the molecular pathogenic mechanisms and treatment strategies. AB - Oculopharyngeal muscular dystrophy (OPMD) is an adult-onset disorder characterized by progressive eyelid drooping, swallowing difficulties and proximal limb weakness. OPMD is caused by a small expansion of a short polyalanine tract in the poly (A) binding protein nuclear 1 protein (PABPN1). The mechanism by which the polyalanine expansion mutation in PABPN1 causes disease is unclear. PABPN1 is a nuclear multi-functional protein which is involved in pre mRNA polyadenylation, transcription regulation, and mRNA nucleocytoplasmic transport. The distinct pathological hallmark of OPMD is the presence of filamentous intranuclear inclusions (INIs) in patient's skeletal muscle cells. The exact relationship between mutant PABPN1 intranuclear aggregates and pathology is not clear. OPMD is a unique disease sharing common pathogenic features with other polyalanine disorders, as well as with polyglutamine and dystrophic disorders. This chapter aims to review the rapidly growing body of knowledge concerning OPMD. First, we outline the background of OPMD. Second, we compare OPMD with other trinucleotide repeat disorders. Third, we discuss the recent advances in the understanding of the molecular mechanisms underlying OPMD pathogenesis. Finally, we review recent therapeutic strategies for OPMD. PMID- 17110090 TI - Water-soluble inhibitor on microbiologically influenced corrosion in diesel pipeline. AB - The effect of water-soluble corrosion inhibitor on the growth of bacteria and its corrosion inhibition efficiency were investigated. Corrosion inhibition efficiency was studied by rotating cage test and flow loop techniques. The nature of biodegradation of corrosion inhibitor was also analyzed by using Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance spectroscopy (NMR) and Gas chromatography and mass spectrometer (GC-MS). The bacterial isolates (Serratia marcescens ACE2, Bacillus cereus ACE4) have the capacity to degrade the aromatic and aliphatic hydrocarbon present in the corrosion inhibitor. The degraded products of corrosion inhibitor and bacterial activity determine the electrochemical behaviour of API 5LX steel. The influence of bacterial activity on degradation of corrosion inhibitor and its influence on corrosion of API 5LX have been evaluated by employing weight loss techniques and electrochemical studies. The main finding of this paper is that the water-soluble corrosion inhibitor is consumed by the microbial action, which contributes to the decrease in inhibitor efficiency. The present study also emphasis the importance of evaluation of water-soluble corrosion inhibitor in stagnant model (flow loop test) and discusses the demerits of the water-soluble corrosion inhibitors in petroleum product pipeline. PMID- 17110091 TI - Binding of cetylpyridinum chloride to glucose oxidase. AB - The binding of cetylpyridinum chloride (CPC) with glucose oxidase (GOD) has been extensively studied at various experimental conditions such as ionic strength, urea concentration and pH at 25 degrees C, using ion-selective membrane electrodes, UV-vis absorption spectroscopy and enzyme activity assay method. The accurate binding isotherms have been obtained and analyzed in terms of Scatchard plot and binding capacity concept. The results represent two binding set system for most of studied conditions. The values of Hill equation parameters have been estimated and used for calculation of intrinsic Gibbs free energy of binding. The results have been interpreted in terms of structural viewpoint of GOD and nature of interactions in the solution. The interpretations are in good agreement with denaturation experiment. Activity measurements represent the significant activation of enzyme due to binding of first CPC molecules. However, the binding of subsequent CPC diminished the activity of enzyme which may be due to the binding of second CPC to enzyme active site. The complete deactivation of enzyme is reached due to binding of about five CPC ions. PMID- 17110092 TI - The link between abdominal obesity, metabolic syndrome and cardiovascular disease. AB - AIM: The prevalence of metabolic syndrome has increased dramatically in recent years, and the cluster of metabolic abnormalities it encompasses results in increased cardiovascular morbidity and mortality. The role of abdominal (visceral) obesity and the underlying molecular and cellular mechanisms central to this association have been the subject of intensive research in recent times. The aim of this review is to correlate data in this area, highlighting the central role of excess visceral fat and its secreted adipokines, and to review existing and emerging therapies. DATA SYNTHESIS: Data were generated from a search of the PubMed database using the terms 'abdominal obesity', 'metabolic syndrome', 'insulin resistance', 'adipokines', 'interleukin-6 (IL-6)', 'adiponectin', 'tumour necrosis factor-alpha (TNF-alpha)' and 'cardiovascular disease'. CONCLUSION: Metabolic syndrome is associated with a pro-inflammatory state, and the role of visceral obesity is thought to be central to this. Visceral obesity leads to alteration of the normal physiological balance of adipokines, insulin resistance, endothelial dysfunction and a pro-atherogenic state. In association with this, the presence of conventional cardiovascular risk factors such as hypertension, dyslipidaemia and smoking results in a significantly elevated cardiovascular and metabolic (cardiometabolic) risk. Better understanding of the molecular mechanisms central to this association has led to the development of potential therapeutic agents. PMID- 17110093 TI - Self/nonself discrimination at the basis of chordate evolution: limits on molecular conservation. AB - All organisms rely on their capacity of self/nonself discrimination to rapidly detect approaching allogeneic cells as well as invading pathogenic microbes as foreign and to eliminate them. Failure to recognize nonself causes self-mating, germline parasitism and disease. Recent findings indicate that, in urochordates - the closest living relatives of vertebrates - different species use completely different molecules for allorecognition. Thanks to their phylogenetic position, these organisms might help us to understand the evolutionary origin of the vertebrate immune system. PMID- 17110094 TI - Factors and values of willingness to pay for improved construction waste management--a perspective of Malaysian contractors. AB - Malaysia is facing an increase in the generation of waste and of accompanying problems with the disposal of this waste. In the last two decades, extensive building and infrastructure development projects have led to an increase in the generation of construction waste material. The construction industry has a substantial impact on the environment, and its environmental effects are in direct relation to the quality and quantity of the waste it generates. This paper discusses general characteristics of the construction contractors, the contractors' willingness to pay (WTP) for improved construction waste management, determining factors which affect the amount of their willingness to pay, and suggestions and policy implications in the perspective of construction waste management in Malaysia. The data in this study is based on contractors registered with the construction industry development board (CIDB) of Malaysia. Employing the open ended contingent valuation method, the study assessed the contractors' average maximum WTP for improved construction waste management to be RM69.88 (1US$=3.6 RM) per tonne of waste. The result shows that the average maximum WTP is higher for large contractors than for medium and small contractors. The highest average maximum WTP value is RM88.00 for Group A (large contractors) RM78.25 for Group B (medium-size contractors) and RM55.80 for Group C (small contractors). One of the contributions of this study is to highlight the difference of CIDB registration grade in the WTP for improved construction waste management. It is found that contractors' WTP for improved waste collection and disposal services increases with the increase in contractors' current paid up capital. The identified factors and determinants of the WTP will assist the formulation of appropriate policies in addressing the construction waste problem in Malaysia and indirectly improve the quality of construction in the country. PMID- 17110095 TI - Chemical and biological characterization of slaughterhouse wastes compost. AB - The chemical and biological properties of compost made from yard trimmings (YT) composted alone or mixed with slaughterhouse wastes (SHW) were evaluated in seven phases. Mixtures were weighed in a 2:1 proportion (YT:SHW) and placed in composting bins (0.91 m2). Temperature was recorded to determine the time (d) needed to reach the first (1HC) and second heat cycles (2HC). Composting characteristics were measured at 0 d, at the peak of the 1HC and 2HC, and at maturation (0, 20, 50 and 70 d). During 1HC, bacterial isolates were cultivated in both treatments and identified using the Biolog System. Chemical composition was statistically analyzed using a 2 (layers of SHW)x7 (composting phases) factorial arrangement of treatments with the ANOVA procedure of SAS. The pH was neutral for YT and ranged from 7.41 to 6.82 for SHW throughout the process. There was a decrease in organic matter (OM) and carbon (C), and a relative increase in nitrogen (N) in both treatments. At 70 d of maturation, C:N values were similar between treatments, but lower (P>0.05) than the initial values. Final N concentration was higher (P>0.05) for the treatment with SHW. Only the SHW treatment exhibited thermophilic temperatures. At the 1HC in both treatments, different populations of bacteria responsible for the breakdown of OM were identified showing an active heterogeneous population. The presence of pathogenic microorganisms was not detected in treatments containing SHW. PMID- 17110096 TI - Label-free immunosensor for prostate-specific antigen based on single-walled carbon nanotube array-modified microelectrodes. AB - We have fabricated a label-free electrochemical immunosensor using microelectrode arrays modified with single-walled carbon nanotubes (SWNTs). Label-free detection of a cancer marker, total prostate-specific antigen (T-PSA), was carried out using differential pulse voltammetry (DPV). The current signals, derived from the oxidation of tyrosine (Tyr), and tryptophan (Trp) residues, increased with the interaction between T-PSA on T-PSA-mAb covalently immobilized on SWNTs. The selectivity of our biosensor was challenged using bovine serum albumin (BSA) as the target protein. The detection limit for T-PSA was determined as 0.25 ng/mL. Since the cut-off limit of T-PSA between prostate hyperplasia and cancer is 4 ng/mL, the performance of our label-free electrochemical immunosensor seems promising for further clinical applications. PMID- 17110097 TI - The prognostic value of proliferation in lymph-node-negative breast cancer patients is age dependent. AB - In lymph-node-negative invasive breast cancer patients<55 years, the proliferation marker mitotic activity index (MAI) has previously been shown to be the strongest prognosticator. In studies without age definition, MAI was not strongly prognostic. We investigated the age dependency of the prognostic value of proliferation for distant metastasis-free (MFS) and overall cancer-related survival (OS) in 1004 histologically diagnosed T1-3N0M0 invasive breast cancers (n=516, <55 years; n=322, 55-70 years; n=166, >70 years) without systemic adjuvant therapy and long follow-up (median: 108 months). The MAI decreases with age and the prognostic value of MAI varied by age group. For patients<55 years, hazard ratios (HR) for MAI>or=10 versus<10 for MFS and OS were 3.1 and 4.4, respectively (P<.0001 for both), but only 1.9 and 1.9 (P=.004 and .006) for patients aged 55-70 years, while over 70 years, MAI was not significant (P=.11). The prognostic value of proliferation was age-dependent. Prognostic breast cancer studies must clearly indicate the age group being studied. PMID- 17110098 TI - Intraparticle diffusion processes during acid dye adsorption onto chitosan. AB - The adsorption of five acid dyes onto chitosan was studied. The equilibrium capacities based on the Langmuir analysis were 1.54, 2.66, 1.11, 1.25 and 1.03 mmol/g chitosan for Orange 10 (AO10), Acid Orange 12 (AO12), Acid Red 18 (AR18), Acid Red 73 (AR73) and Acid Green 25 (AG25) respectively. The batch adsorption rate for the five systems based on an intraparticle diffusion rate parameter derived from the plots of dye adsorbed versus the square root of time indicated that the adsorption mechanism was predominantly intraparticle diffusion but there was also a dependence on pore size as the dye diffuses through macropore, mesopore and micropore respectively. PMID- 17110099 TI - Contaminant occurrence, identification and control in a pilot-scale corn fiber to ethanol conversion process. AB - While interest in bioethanol production from lignocellulosic feedstocks is increasing, there is still relatively little pilot-plant data and operating experience available for this emerging industry. A series of batch and continuous fermentation runs were performed in a pilot-plant, some lasting up to six weeks, in which corn fiber-derived sugars were fermented to ethanol using glucose fermenting and recombinant glucose/xylose-fermenting yeasts. However, contamination by Lactobacillus bacteria was a common occurrence during these runs. These contaminating microorganisms were found to readily consume arabinose, a sugar not utilized by the yeast, producing acetic and lactic acids that had a detrimental effect on fermentation performance. The infections were ultimately controlled with the antibiotic virginiamycin, but routine use of antibiotics is cost prohibitive. The severity of the problem encountered during this work is probably due to use of a highly contaminated feedstock. Lignocellulosic conversion facilities will not employ aseptic designs. Instead, techniques similar to those employed in the corn-based fuel ethanol industry to control infections will be used. Effective control may also be possible by using fermentative microorganisms that consume all biomass-derived sugars. PMID- 17110100 TI - Solid phase bioremediation of pendimethalin in contaminated soil and evaluation of leaching potential. AB - Substrate leaching experiments were performed to study the relative leaching potential of pendimethalin in various types of soil matrices. Pendimethalin leaching showed up to a depth of 30 cm in all the studied soil matrices, irrespective of pH conditions used. The leaching potential of pendimethalin was assessed at various pH conditions. Comparatively higher leaching potential was observed in basic conditions compared to the neutral and acid conditions of soil. Soil phase bioremediation of pendimethalin was also performed on all the soil matrices. Among the studied variations, bioremediation experiments performed in presence of sunlight showed higher efficiency. Bioaugmentation along with sunlight showed higher remediation efficiency in all the studied soil matrices. Biostimulation did not respond positively on the progress of bioremediation. PMID- 17110101 TI - Recovery of antioxidant phenolics from white vinification solid by-products employing water/ethanol mixtures. AB - Solid wastes from white vinification, including grape peels, seeds and stems, were used as raw material for the recovery of antioxidant polyphenols. Extractions were performed using non-toxic media composed of water/ethanol mixtures and hydrochloric, acetic or tartaric acid. Recovery efficiency was assessed by monitoring the antioxidant potency of extracts and several indices related to their polyphenolic composition, including total polyphenol, total flavonoid, total flavanol and condensed tannin (proanthocyanidin) content. Among the by-products tested, seeds were shown to contain exceptional amounts of total polyphenols (13.76 g per 100g dry weight), followed by stems (7.47 g per 100g dry weight) and peels (0.97 g per 100g dry weight). Extracts with the highest antioxidant activity from all by-products were obtained with 57% ethanol. Acidification of this medium with 0.1% HCl improved polyphenol recovery and antiradical activity for stem extracts, but it was unfavourable for seed extraction. PMID- 17110102 TI - The kinetics of the removal of nitrogen and organic matter from parboiled rice effluent by cyanobacteria in a stirred batch reactor. AB - The aim of this research was to evaluate the kinetics of the removal of nitrogen and organic matter from parboiled rice effluent by the cyanobacteria Aphanothece microscopica Nageli. From the results obtained, it was verified that maximum efficiency for the removal of organic matter expressed as COD and total nitrogen (N-TKN) occurred after 15 h of cultivation, being 83.44% and 72.74, respectively. The scale-up process indicated that the volume of the estimated reactor would be 100.2m(3), containing parboiled rice effluent and biomass. PMID- 17110103 TI - Synthesis of a technetium-99m labeled tricyclic ganciclovir analog for non invasive reporter gene expression imaging. AB - A potential radiopharmaceutical and HSV1-TK substrate, 3-((1,3-dihydroxypropan-2 yloxy)-methyl)-6-(4-(3-((2-mercaptoethyl)(2-(2-mercaptoethyl-amino) ethyl)amino)propoxy)phenyl)-3H-imidazopurin-9(5H)-one-oxo-technetium(V), was synthesized via a converging approach and its chemical structure was comparatively characterized with a non-radioactive analog. The final radiochemical purity and yield were 97 and 73%, respectively. PMID- 17110104 TI - Trifunctional norrisolide probes for the study of Golgi vesiculation. AB - Inspired by the effect of norrisolide on the Golgi complex, we synthesized norrisolide probes that contain: the perhydroindane core of the parent natural product for Golgi localization, a crosslinking unit (aryl azide or epoxide) for covalent binding to the target, and a tag (biotin or iodine) for subsequent target purification. We found that biotin-containing probes 14, 20 and 24 induced inefficient Golgi vesiculation. However, the iodinated probe 25 induced extensive and irreversible Golgi fragmentation. This probe can be used for the isolation of the cellular target of norrisolide. PMID- 17110105 TI - Synthesis and biological activities of novel dexibuprofen tetraacetylriboflavin conjugates. AB - A series of novel dexibuprofen derivatives covalently linked via alkylene spacers of variable length to tetraacetylated riboflavin have been developed. The target compounds became accessible by reaction of the chloromethyl ester of dexibuprofen with tetraacetylriboflavin (compound 7) or by synthesis of the appropriate N3 (omega-iodoalkyl)-2',3',4',5'-Tetraacetylriboflavin followed by treatment with dexibuprofen (derivatives 8-11), respectively. Biological screening revealed that the target compounds exhibit antiproliferative effects on MCF-7 breast cancer and HT-29 colon carcinoma cells with IC50 values in the range of 8-15 microM. Enzymatic studies on human platelets indicated significant COX-1 inhibitory activities of the target compounds. PMID- 17110106 TI - Discovery of adamantane ethers as inhibitors of 11beta-HSD-1: Synthesis and biological evaluation. AB - A novel class of adamantane ethers 11beta-hydroxysteroid hydrogenase type I inhibitors has been discovered. These compounds have excellent HSD-1 potency and selectivity against HSD-2. The structure-activity relationships, selectivity, metabolism, PK, ex vivo pharmacodynamic data, and an X-ray crystal structure of one of these inhibitors bound to h-HSD-1 are discussed. PMID- 17110107 TI - Synthesis and antiproliferative activity of (2R,3R)-disubstituted tetrahydropyrans. Part 2: Effect of side chain homologation. AB - In this study, we synthesized a series of enantiomerically pure (2R,3R)- and (2R,3S)-disubstituted tetrahydropyrans bearing a CH2O spacer group on the side chain at position 2 of the heterocyclic ring. The in vitro antiproliferative activities of the compounds were examined in the human solid tumor cell lines A2780 (ovarian cancer), SW1573 (non-small cell lung cancer), and WiDr (colon cancer). Overall, the results point out the relevance for antiproliferative activity of the distance between the heterocycle and the unsaturated group. PMID- 17110108 TI - An efficient synthesis of quinoxaline derivatives from 4-chloro-4-deoxy-alpha-D galactose and their cytotoxic activities. AB - A novel and efficient method for the synthesis of quinoxaline derivatives has been developed. Isopropylidenation of 4-chloro-4-deoxy-alpha-D-galactose with 2,2 dimethoxypropane, followed by selective hydrolysis, afforded 2,3-O-isopropylidene 4-chloro-4-deoxy-D-galactose di-methyl acetal (3) as a sole product. Oxidation of compound 3 with (Bu3Sn)2O-Br2 gave corresponding hex-5-ulose derivative in high yields. The hex-5-ulose derivative reacted with o-phenylenediamines under neutral conditions to afford quinoxaline derivatives in reasonable yields. The in vitro cytotoxic activities of these quinoxaline derivatives were investigated. PMID- 17110109 TI - Remarkable immunostimulation effects of hybrid liposomes on human peripheral blood mononuclear cells in vitro. AB - Immunostimulation effects of hybrid liposomes (HL25) composed of 90 mol% l-alpha dimyristoylphosphatidylcholine (DMPC) and 10 mol% polyoxyethylene(25)dodecyl ether (C12(EO)25) on the normal human peripheral blood mononuclear cells (hPBMCs) were investigated in vitro. The concentrations of cytokines (IFN-gamma, IL-12) were significantly increased in the presence of HL25 ([DMPC] = 100 microM, [C12(EO)25] = 10 microM) and the maximum values attained were 13-14 times higher compared with those of control, though the viability and proliferation of hPBMCs were decreased under the same conditions. Such a remarkably high cytokine production induced by liposomes without any stimulators was obtained for the first time. Furthermore, fluorescence microscopic and flow cytometric analyses indicated that the HL25 could be incorporated into hPBMCs by the membrane fusion and/or the endocytosis. These results suggest that HL25 should induce the effective immunopotentiation by stimulating the cytokine productivity of hPBMCs. PMID- 17110110 TI - Synthesis and biological study of medicinally important Mannich bases derived from 4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,6,10,12,12a pentahydroxy naphthacene carboxamide. AB - The paper describes synthesis and antibacterial study of biologically active Mannich bases of carboxamide derivative employing Mannich reaction of 4 (dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,6,10,12,12a pentahydroxy naphthacene carboxamide with various sulfonamides/secondary amines . They were analysed by elemental analysis and characterized by UV, IR and (1)H NMR spectroscopic studies. The Mannich bases were screened for antibacterial activity against various gram-negative bacteria at various concentrations and were analysed statistically. The result has shown that the compounds are quite active against pathogens under study and were non-toxic. All the synthesized compounds were found to be low lethal as ascertained by LD50 test. PMID- 17110111 TI - Anti-influenza virus activity of biflavonoids. AB - Ginkgetin was found to inhibit the influenza virus sialidase. Ginkgetin-sialic acid conjugates showed a significant survival effect in the influenza-virus infected mice. PMID- 17110112 TI - Preliminary normative values for foot loading parameters of the developing child. AB - The purpose of the present study was to provide normative data for foot loading patterns and foot shape parameters in order to support decisions about the normal or abnormal development of the growing foot during childhood. In a longitudinal design, 90 healthy German children were followed over the course of 6-48 months. The children had a mean age of 15 months at the first appointment and 63 months at the last appointment. The children participated in 11 measurement appointments every 3 months or 6 months. Dynamic foot loading was evaluated with plantar pressure measurements during walking and static footprints were taken to determine changes in foot shape. During the investigation period a significant increase of peak pressure in almost every foot region was observed. Peak pressure of the total foot increased by 92%. Only for the midfoot region a significantly decreased impulse by about 15% per year and a significant decrease of the contact area by 9% per year could be observed. A significant influence of gender was observed for peak pressures and impulses in some foot regions as well as for the midfoot width and the foot shape index. The established database can be used as comparative values for clinical decisions about the normal development. PMID- 17110113 TI - Synthesis and biological evaluation of several structural analogs of 2 arachidonoylglycerol, an endogenous cannabinoid receptor ligand. AB - 2-Arachidonoylglycerol (2-AG (1)) is an endogenous ligand for the cannabinoid receptors (CB1 and CB2). There is growing evidence that 2-arachidonoylglycerol plays important physiological and pathophysiological roles in various mammalian tissues and cells, though the details remain to be clarified. In this study, we synthesized several remarkable analogs of 2-arachidonoylglycerol, closely related in chemical structure to 2-arachidonoylglycerol: an analog containing an isomer of arachidonic acid with migrated olefins (2-AGA118 (3)), an analog containing a one-carbon shortened fatty acyl moiety (2-AGA113 (4)), an analog containing an one-carbon elongated fatty acyl moiety (2-AGA114 (5)), a hydroxy group-containing analog (2-AGA105 (6)), a ketone group-containing analog (2-AGA109 (7)), and a methylene-linked analog (2-AGA104 (8)). We evaluated their biological activities as cannabinoid receptor agonists using NG108-15 cells which express the CB1 receptor and HL-60 cells which express the CB2 receptor. Notably, these structural analogs of 2-arachidonoylglycerol exhibited only weak agonistic activities toward either the CB1 receptor or the CB2 receptor, which is in good contrast to 2-arachidonoylglycerol which acted as a full agonist at these cannabinoid receptors. These results clearly indicate that the structure of 2 arachidonoylglycerol is strictly recognized by the cannabinoid receptors (CB1 and CB2) and provide further evidence that the cannabinoid receptors are primarily the intrinsic receptors for 2-arachidonoylglycerol. PMID- 17110114 TI - C6-(N,N-butyl-methyl-heptanamide) derivatives of estrone and estradiol as inhibitors of type 1 17beta-hydroxysteroid dehydrogenase: Chemical synthesis and biological evaluation. AB - A series of estrone and estradiol derivatives having an N-butyl,methyl heptanamide side chain at C6-position were synthesized, tested as inhibitors of type 1 17beta-HSD and assessed for their possible estrogenic activity. A better type 1 17beta-HSD inhibition was obtained for the 6beta-side chain orientation over 6alpha; the C17-alcohols are more potent inhibitors than the corresponding ketones; introducing a 2-methoxy group decreased the inhibitory potency; and the replacement of a C-S bond by a C-C bond in the C6beta-side chain is not detrimental to inhibition. Interestingly, the new inhibitors were also found less estrogenic than the lead compound in two breast cancer cell lines, T-47D and MCF 7. PMID- 17110115 TI - ABP688, a novel selective and high affinity ligand for the labeling of mGlu5 receptors: identification, in vitro pharmacology, pharmacokinetic and biodistribution studies. AB - [(11)C]ABP688 (2) has recently been demonstrated to be a useful PET tracer for in vivo imaging of the metabotropic glutamate receptors type 5 (mGluR5) in rodents. We describe here the identification and preclinical profiling of ABP688 and its tritiated version [(3)H]ABP688, and show that its high affinity (K(d)=2nM), selectivity, and pharmacokinetic properties fulfill all requirements for development as a PET tracer for clinical imaging of the mGlu5 receptor. PMID- 17110116 TI - Structure-activity relationships of 19-norvitamin D analogs having a fluoroethylidene group at the C-2 position. AB - We have synthesized four new geometric isomers of 1alpha,25-dihydroxy-2-(2' fluoroethylidene)-19-norvitamin D analogs 1 and 2 having a 20R- and 20S configuration, whose structures are correlated with 2MD possessing high potencies in stimulating bone formation in vitro and in vivo. The E-isomers of (20R)- and (20S)-2-fluoroethylidene analogs 1a and 1b were comparable with the natural hormone 1alpha,25-(OH)(2)D(3) in binding to the vitamin D receptor (VDR), while two Z-isomers 2a and 2b were about 15-20 times less active than the hormone. In inducing expression of the vitamin D responsive element-based luciferase reporter gene, the E-isomers 1a and 1b were 1.2- and 8.6-fold more potent than the hormone, respectively, while the Z-isomers 2a and 2b had 27-55% of the potency. On the basis of the biological activities and a docking simulation based on X-ray crystallographic analysis of the VDR ligand-binding pocket, the structure activity relationships of the fluorinated 19-norvitamin D analogs are discussed. PMID- 17110117 TI - TOMOCOMD-CARDD descriptors-based virtual screening of tyrosinase inhibitors: evaluation of different classification model combinations using bond-based linear indices. AB - A new set of bond-level molecular descriptors (bond-based linear indices) are used here in QSAR (quantitative structure-activity relationship) studies of tyrosinase inhibitors, for finding functions that discriminate between the tyrosinase inhibitor compounds and inactive ones. A database of 246 compounds was collected for this study; all organic chemicals were reported as tyrosinase inhibitors; they had great structural diversity. This dataset can be considered as a helpful tool, not only for theoretical chemists but also for other researchers in this area. The set used as inactive has 412 drugs with other clinical uses. Twelve LDA-based QSAR models were obtained, the first six using the non-stochastic total and local bond-based linear indices as well as the last six ones, the stochastic molecular descriptors. The best two discriminant models computed using the non-stochastic and stochastic molecular descriptors (Eqs. , respectively) had globally good classifications of 98.95% and 89.75% in the training set, with high Matthews correlation coefficients (C) of 0.98 and 0.78. The external prediction sets had accuracies of 98.89% and 89.44%, and (C) values of 0.98 and 0.78, for models 7 and 13, respectively. A virtual screening of compounds reported in the literature with such activity was carried out, to prove the ability of present models to search for tyrosinase inhibitors, not included in the training or test set. At the end, the fitted discriminant functions were used in the selection/identification of new ethylsteroids isolated from herbal plants, looking for tyrosinase inhibitory activity. A good behavior is shown between the theoretical and experimental results on mushroom tyrosinase enzyme. It might be highlighted that all the compounds showed values under 10microM and that ES2 (IC(50)=1.25microM) showed higher activity in the inhibition against the enzyme than reference compounds kojic acid (IC(50)=16.67microM) and l-mimosine (IC(50)=3.68microM). In addition, a comparison with other established methods was carried to prove the adequate discriminatory performance of the molecular descriptors used here. The present algorithm provided useful clues that can be used to speed up in the identification of new tyrosinase inhibitor compounds. PMID- 17110118 TI - Scintigraphic imaging with a peptide glucuronide in rabbits: 99mTc- exorphin C glucuronide. AB - A peptide glucuronide (Exorphin C glucuronide) was labeled with 99mTc using glucoheptonate (GH) as a bifunctional chelating agent. Scintigraphic imaging was performed in male Albino rabbits. Exorphin C glucuronide showed rapid and efficient labeling with 99mTc using glucoheptonate as a bifunctional chelate. Results demonstrated that 99mTc-GEG may be a useful new type of glucuronide derivative of peptides for diagnosis of some cancer diseases. PMID- 17110119 TI - On the preparation of a therapeutic dose of 177Lu-labeled DOTA-TATE using indigenously produced 177Lu in medium flux reactor. AB - 177Lu could be produced with a specific activity of approximately 23,000 mCi/mg (850GBq/mg) by neutron activation using enriched 176Lu (64.3%) target when irradiation was carried out at a thermal neutron flux of 1 x 10(14) n/cm(2)/s for 21 d. 177Lu-DOTA-TATE could be prepared in high radiochemical yield (approximately 99%) and adequate stability using the 177Lu produced indigenously. The average level of radionuclidic impurity burden in 177Lu due to 177mLu was found to be 250 nCi of 177mLu/1 mCi of 177Lu (9.25 kBq/37 MBq) at the end of bombardment, which corresponds to 0.025% of the total activity produced. The maximum specific activity achievable via careful optimization of the irradiation parameters was found to be adequate for the preparation of a therapeutic dose of the radiopharmaceutical. The in-house preparation of this agent using 25 microg (17.41 nmole) of DOTA-TATE and indigenously produced 177Lu (0.8 microg, 4.52 nmole), corresponding to peptide/Lu ratio of 3.85 yielded 98.7% complexation. Allowing possibility of decay due to transportation to users, it has been possible to demonstrate that at our end, a single patient dose of 150-200 mCi (5.55-7.40 GBq) can be prepared by using 250-333 microg of DOTA-TATE conjugate. This amount compares well with 177Lu-DOTA-TATE prepared for a typical peptide receptor radionuclide therapy (PRRT) procedure which makes use of 100 microg of the DOTA-TATE conjugate, which incorporates 50 mCi (1.85 GBq) of 177Lu activity, thereby implying that in order to achieve a single patient dose of 150-200 mCi (5.55-7.40 GBq), 300-400 microg of the conjugate needs to be used. PMID- 17110120 TI - Transcriptional profiling of cell wall protein genes in chickpea embryonic axes during germination and growth. AB - Cell wall hydrolases have been assumed to be involved in the regulation of seed germination, mostly through their contribution to the cell wall disassembly associated with endosperm cap weakening. In Cicer arietinum (a non-endospermic leguminosae seed), we have focused our research directly on the elongation process of the embryonic axes themselves during germination. The genes encoding cell wall proteins, previously implicated in the elongation of chickpea epicotyls, might also be involved in the expansion of embryonic axis cells, and the modulation of their expression could be part of the control of the germinative process. Thus, chickpea alpha-expansins and xyloglucan endotransglycosylase/hydrolase (XTH) acting on the cellulose/xyloglucan network seem to be involved in the elongation of both chickpea epicotyls and embryonic axes, although the products of different genes perform their actions on each organ. Among the four known cDNAs encoding chickpea alpha-expansins, Ca-EXPA1 was the only isoform highly expressed in embryonic axes during germination. In contrast to epicotyl elongation, the genes encoding cell wall beta galactosidases, involved in pectin degradation, were not expressed during germination, suggesting no role in embryonic axis elongation, mainly due to the different metabolism of pectins during cell wall loosening in embryonic axis or epicotyl cells. The results concerning CanST-1 and -2, encoding two growth related cell wall proteins, suggested that these genes were not involved in elongation of embryonic axes during germination. The transcription pattern of Cap28, which encodes a glutamic acid rich cell wall protein of unknown function, indicated a role in the development of the embryonic axes during germination. PMID- 17110121 TI - Cloning and characterization of six highly similar endo-1,3-beta-glucanase genes in hexaploid wheat. AB - Pathogenesis-related (PR) proteins are often used as a marker of plant defense reactions. Some endo-1,3-beta-glucanase (Gns) genes encode proteins that belong to the PR protein family 2 (PR-2). Although the number of homologous family member genes is significantly greater in hexaploid wheat (Triticum aestivum L.) compared to other model plants, earlier studies did not evaluate the possible contribution of their homologs to hybridization signals in Northern blot analysis. In this study, we have examined whether routine transcriptional analyses of a PR gene is of high reliability or not by isolating six highly similar Gns genes (TaGlb2a, TaGlb2b, TaGlb2c, TaGlb2d, TaGlb2e, and TaGlb2f) and characterizing their expression patterns in detail. While TaGlb2b was shown to be a PR-2 gene, transcription of TaGlb2c and TaGlb2d was not induced upon infection with either powdery mildew (Erysiphe graminis) or head blight (Fusarium graminearum) pathogens; their transcripts were most abundant in healthy spikes (lemmas and in particular paleae). Therefore, in some cases, the conventional analyses do not necessarily provide accurate information on expression pattern of a PR gene in hexaploid wheat. This is also the first report of wheat genes that are specifically expressed in lemma/palea tissues of flowering spikelets. PMID- 17110122 TI - Accumulation of tocopherols and tocotrienols during seed development of grape (Vitis vinifera L. cv. Albert Lavallee). AB - Tocopherols and tocotrienols are present in mature seeds. Yet, little is known about the physiological role and the metabolism of these compounds during seed development. Here we present data on tocopherol and tocotrienol accumulation during seed development in Vitis vinifera L. cv. Albert Lavallee (Royal). This species was chosen for its ability to synthesize both tocopherols and tocotrienols. It is shown here for the first time that during seed development there are significant differences in localization and accumulation kinetics of tocopherols and tocotrienols. Tocopherols are found homogeneously dispersed throughout all tissues of the seed, in concentrations ranging from 20 to 100 microg tocopherol per g dry weight. Tocopherol levels decrease gradually during seed development. In contrast, tocotrienols are only found in the endosperm of the seeds, accumulating in a sigmoid fashion during the maturation period of seed development. Tocotrienol levels were found to be (54+/-7.4) microg/g dry seed in 90-day-old seeds of V. vinifera L. Furthermore, tocotrienol biosynthesis is demonstrated in these seeds during tocotrienol accumulation and in an endosperm fraction isolated at 75 days after flowering. PMID- 17110123 TI - One-stage neonatal corrective repair for d-transposition of the great arteries and complete atrio-ventricular canal. AB - Association of d-transposition of the great arteries and complete atrio ventricular canal constitutes an uncommon and complex cardiac anomaly usually associated with poor prognosis. We report our experience on one-stage neonatal repair for d-transposition of the great arteries and complete atrio-ventricular canal. Between August 1997 and 2005, four patients (two males and two females) underwent anatomical correction for d-transposition of the great arteries and complete atrio-ventricular canal using an arterial switch procedure and two-patch repair. Mean age and weight at operation were 20 days (range from 3 to 28 days) and 3.2kg (range from 2.7 to 3.5kg), respectively. None of the patients received preoperative palliative procedure. Associated lesions were left outflow tract obstruction in three patients and multiple muscular ventricular septal defects in two patients. All four patients survived the operation. There was one in hospitality death due to fungal sepsis. One patient required late re-operation for left ventricular outflow tract obstruction and left atrio-ventricular valve regurgitation. For a mean follow-up of 67 months (range from 51 to 90 months) all patients are asymptomatic and with no residual defects. Corrective repair of d transposition of the great arteries and complete atrio-ventricular canal can be successfully achieved in this very challenging population during the neonatal period. PMID- 17110124 TI - Does adjuvant radiation therapy improve disease-free survival in completely resected Masaoka stage II thymoma? AB - OBJECTIVE: To determine whether or not patients with completely resected Masaoka stage II thymoma benefit from postoperative radiotherapy (RT). METHODS: We retrospectively review the case records and compared the long-term outcomes of patients affected by Masaoka stage II thymoma treated by resection alone with same stage thymoma patients submitted to resection and RT. Surgical specimens were reviewed to confirm pathological stage, negative resection margins and histological subtype. RESULTS: Between 1988 and 2000, we performed 197 resections for thymoma; 58 patients resulted to be affected by completely resected tumours with microscopic transcapsular invasion (stage IIA, n=25) or macroscopic invasion into the surrounding fatty tissue with or without adhesion to the mediastinal pleura (stage IIB, n=33). Thirty-two patients underwent only complete surgical resection (14 stage IIA and 18 stage IIB); 26 patients underwent complete resection and subsequent mediastinal RT (11 stage IIA and 15 stage IIB). RT dosages were 45-54grays (Gy), in 25-30 fractions. Histological subtypes were similarly represented in both groups. Median follow-up was 91 months (range 9 170). Five intrathoracic recurrences occurred: three radiated patients (2 stage IIB - 1 AB and 1 B2 thymoma; 1 stage IIA B1 thymoma) and two not-radiated patients (1 stage IIA AB thymoma and 1 stage IIB B1 thymoma). Disease-free survival rate at 5- and 10-year were 94% and 87%, respectively. Log-rank test showed no difference in Kaplan-Meier survival curves (p=0.432) between radiated and not-radiated patients. CONCLUSIONS: These data support the concept that radical surgical resection alone should be considered a sufficient treatment for stage II thymoma. PMID- 17110125 TI - Activity of Subtilisin Carlsberg in macromolecular crowding. AB - Enzymatic activity of a proteolytic enzyme Subtilisin Carlsberg (SC) in anionic sodium dodecyl sulfate (SDS) micellar medium has been explored and found to be retarded compared to that in bulk buffer. Circular dichroism (CD) study reveals that SDS, which is a potential protein denaturant, has an insignificant denaturation effect on SC. The structural integrity of the protein offers an opportunity to study the functionality of the enzyme SC in a macromolecular crowding of micelles. Dynamic light scattering (DLS) data indicates no sandwich like micelle-SC complex formation ruling out the possibility of interaction of the enzyme with the hydrophobic core of the micelle. However, steady state and time resolved emission studies on specific and nonspecific fluorescent probes indicate the proximity effect at the surface of the enzyme due to macromolecular crowding of the micelles. The agreement of retarded enzymatic activity in the micellar crowd with a theoretical model ascribed to the facts that substrates are compartmentalized in the micelles and enzyme interacts with the micelle through stern layer. PMID- 17110126 TI - Immunological activation of human umbilical cord blood mast cells induces tryptase secretion and interleukin-6, and histidine decarboxilase mRNA gene expression. AB - Allergy is the result of a complex immune cascade leading to the disregulated production of Th2 cytokines, the generation of allergen-specific IgE-producing B cells and the subsequent activation and degranulation of mast cells upon allergen challenge. Mast cell effector function significantly influences the quantity, duration and magnitude of most allergic reactions. Here, using isolated human umbilical cord blood mast cells (HUCBMC) from CD34+ cells, activated with anti IgE (10 microg/ml) in culture, we found an augmented release of IL-6, tryptase and histamine (p < 0.01 compared with control). In addition, in these cells anti IgE (10 microg/ml) activated the expression of histidine decarboxylase (HDC) and IL-6. In these studies we describe a new biological activity of anti-IgE in inducing histidine decarboxylase and IL-6, suggesting that this cytokine may have an important effect on allergic and inflammatory diseases mediated by mast cells. Moreover, with these data we confirm the immunoregulatory and inflammatory function of mast cells. PMID- 17110127 TI - 2-Step purification of the Ku DNA repair protein expressed in Escherichia coli. AB - The Ku protein is involved in DNA double-strand break repair by non-homologous end-joining (NHEJ), which is crucial to the maintenance of genomic integrity in mammals. To study the role of Ku in NHEJ we developed a bicistronic Escherichia coli expression system for the Ku70 and Ku80 subunits. Association of the Ku70 and Ku80 subunits buries a substantial amount of surface area (approximately 9000 A2 [J.R. Walker, R.A. Corpina, J. Goldberg, Structure of the Ku heterodimer bound to DNA and its implications for double-strand break repair, Nature 412 (2001) 607 614]), which suggests that herterodimerization may be important for protein stability. N-terminally His6-tagged Ku80 was soluble in the presence, but not in the absence, of bicistronically expressed untagged Ku70. In a 2-step purification, metal chelating affinity chromatography was followed by step gradient elution from heparin-agarose. Co-purification of equimolar amounts of His6-tagged Ku80 and untagged Ku70 was observed, which indicated heterodimerization. Recombinant Ku bound dsDNA, activated the catalytic subunit of the DNA-dependent kinase (DNA-PKcs) and functioned in NHEJ reactions in vitro. Our results demonstrate that while the heterodimeric interface of Ku is extensive it is nonetheless possible to produce biologically active Ku protein in E. coli. PMID- 17110128 TI - Expression, purification, and characterization of an aminopeptidase (Xac2987) with broad specificity from Xanthomonas axonopodis pv. citri. AB - We report here, the cloning, expression, and purification of a broad specificity aminopeptidase from Xanthomonas campestris pv. citri in fusion with a hexa histidine tag at the N-terminal portion of the protein to facilitate purification. The protein was expressed in the soluble fraction and could be purified in one step by IMAC, yielding approximately 50mg pure protein per liter of cells. We show that the protein is folded and presents aminopeptidase activity against synthetic substrates. Also, we present the characterization of its specificity, showing that the protein was, indeed, able to catalyze the removal of N-terminal residues from synthetic substrates. PMID- 17110129 TI - RAG-1 and IgM genes, markers for early development of the immune system in the gadoid haddock, Melanogrammus aeglefinus, L. AB - The full sequence of the heavy chain of Immunoglobulin M (IgM) and a partial fragment of the recombination activating gene-1 (RAG-1) gene were sequenced in haddock, Melanogrammus aeglefinus, L. The complete transcript of haddock IgM consisted of 1865 bp and translated into a 572-aa peptide. The RAG-1 fragment was 1776 bp and was identified as the core region of RAG-1. These two immune genes were used in expression studies as markers of early development in haddock larvae. A DIG labelled oligoprobe of the RAG-1 gene was used in whole-mount in situ hybridisation (WISH). A hybridisation signal for RAG-1 was first detected in larvae at 25 days post-hatching (dph) in two bilateral symmetric regions of the head identified as the thymus. Further expression studies were carried out by RT PCR analysis of RAG-1 and IgM on larval samples obtained during early development, i.e. from fertilisation to weaning. Haddock RAG-1 expression was detected after 21 days post-fertilisation (dpf) whilst IgM transcripts were not detected until 40 dpf, equivalent to day 29 post-hatching. These results suggest that the immune system in haddock starts to develop in larvae of 6-7 mm in length (25-29 dph). PMID- 17110130 TI - Gastric fundic distension activates fronto-limbic structures but not primary somatosensory cortex: a functional magnetic resonance imaging study. AB - INTRODUCTION: The brain representation of visceral stimulation bears important similarities to that of somatic stimulation. However, the role of the primary (S1) and secondary (S2) somatosensory cortices in mediating gastric sensation is uncertain. MATERIALS AND METHODS: Eighteen healthy, right-handed volunteers (age 32 years+/-6.5 years; 14 men) underwent dynamic assessment of the relationship between sensation and fundic barostat distending pressure and volume, and then brain functional magnetic resonance imaging (fMRI) during noxious fundic distension. Cytoarchitectonic probability maps were used to examine in detail the null hypothesis that fundic distension did not produce significant activation of S1 or S2. RESULTS: Distending volume explained 74% of the variance in gastric sensation, compared to 64% with distending pressure. Incorporating distending volume into the regressor function for our fMRI analyses, we found that noxious fundic distension activated a widespread network of brain regions, including the pontine brainstem, thalami, cerebellum, insular cortex bilaterally, anterior and posterior cingulate cortex, right frontal lobe, and inferior parietal lobules. In detailed analyses, we found no evidence of activation of S1, but did find activation in one region of S2. DISCUSSION: Our findings suggest that an extensive, predominantly fronto-limbic network of brain regions, including the insular cortex, mediates perception of noxious gastric fundic distension in healthy humans, without significant participation by the primary somatosensory cortex. This and other recent studies lay the groundwork for investigations comparing brain processing of visceral stimuli between healthy volunteers and patients with functional dyspepsia. PMID- 17110131 TI - Spatial accuracy of fMRI activation influenced by volume- and surface-based spatial smoothing techniques. AB - As improvements in cortical surface modeling allowed accurate cortical topology in brain imaging studies, surface-based methods for the analysis of functional magnetic resonance imaging (fMRI) were introduced to overcome the topological deficiency of commonly used volume-based methods. The difference between the two methods is mainly due to the smoothing techniques applied. For practical applications, the surface-based methods need to quantitatively validate the accuracy of localizing activation. In this study, we evaluated the spatial accuracy of activation detected by the volume- and surface-based methods using simulated blood oxygenation level-dependent (BOLD) signals and MRI phantoms focusing on the influence of their smoothing techniques. T1- and T2-weighted phantoms were acquired from BrainWeb () and used to extract cortical surfaces and to generate echo planar imaging (EPI) data. Simulated BOLD signals as the gold standard of activation in our experiment were applied to the surfaces and projected to the volume space with random noise. Three-dimensional isotropic Gaussian kernel smoothing and two-dimensional heat kernel smoothing were applied to the volume- and surface-based methods. Sensitivity and 1-specificity, which are truly and falsely detected activations, and similarity measures, which are spatially and statistically similar for the gold standard and detected activations, were calculated. In the results, the surface-based method showed the sensitivity and similarity scores of about 12% higher than the volume-based method. In conclusion, the surface-based method guarantees better spatial accuracy for the localization of BOLD signal sources within the cortex than the volume-based method. PMID- 17110132 TI - Impact of modality and linguistic complexity during reading and listening tasks. AB - Reading and understanding speech are usually considered as different manifestations of a single cognitive ability, that of language. In this study, we were interested in characterizing the specific contributions of input modality and linguistic complexity on the neural networks involved when subjects understand language. We conducted an fMRI study during which 10 right-handed male subjects had to read and listen to words, sentences and texts in different runs. By comparing reading to listening tasks, we were able to show that the cerebral regions specifically recruited by a given modality were circumscribed to unimodal and associative unimodal cortices associated with the task, indicating that higher cognitive processes required by the task may be common to both modalities. Such cognitive processes involved a common phonological network as well as lexico semantic activations as revealed by the conjunction between all reading and listening tasks. The restriction of modality-specific regions to their corresponding unimodal cortices was replicated when looking at brain areas showing a greater increase during the comprehension of more complex linguistic units than words (such as sentences and texts) for each modality. Finally, we discuss the possible roles of regions showing pure effect of linguistic complexity, such as the anterior part of the superior temporal gyrus and the ventro-posterior part of the middle temporal gyrus that were activated for sentences and texts but not for isolated words, as well as a text-specific region found in the left posterior STS. PMID- 17110133 TI - A three-dimensional, histological and deformable atlas of the human basal ganglia. I. Atlas construction based on immunohistochemical and MRI data. AB - This paper describes the construction of an atlas of the human basal ganglia. The successive steps of the construction were as follows. First a postmortem specimen was subjected to a MRI acquisition prior to extraction of the brain from the skull. The brain was then cryosectioned (70 microm thickness). One section out of ten (80 sections) was Nissl-stained with cresyl violet, another series of 80 sections was immunostained for the calcium binding protein calbindin. Contours of basal ganglia nuclei including their calbindin-stained functional subdivisions, fiber bundles and ventricles (n=80 structures) were traced from histological sections and digitized. A novelty of this atlas is the MRI acquisition, which represents the core data element of the study. MRI was used for the coregistration of the atlas data and permitted, through multimodal (Nissl, calbindin, images of cryosectioning, T1 and T2 MRI) and 3D optimization, the production of anatomically and geometrically consistent 3D surfaces, which can be sliced through any desired orientation. The atlas MRI is also used for its deformation to provide accurate conformation to the MRI of living patients, thus adding information at the histological level to the patient's MRI volume. This latter aspect will be presented in a forthcoming paper. PMID- 17110134 TI - Homocysteine and bone loss in epilepsy. AB - Epidemiological studies reveal fracture incidence in epilepsy is twice that of the normal population. Much interest has been focused on Vitamin D, however, considering mixed results on non-enzyme inducing anti-epileptic drugs (AEDs) and bone mineral density (BMD) additional metabolic effects may be to blame. AEDs increase serum homocysteine (s-Hcy) by lowering blood folate levels. An association between elevated homocysteine, BMD and increased fracture incidence has been found in non-epilepsy populations. Additionally, folate and Vitamin B12 levels are independently related to bone mineral density in various non-epilepsy populations. This study supports previous research, which found elevated s-Hcy in subjects taking AEDs and that bone loss is related to the use of enzyme-inducing AEDs and changes in alkaline phosphatase. By one-way ANOVA, subjects on phenytoin monotherapy had significantly higher levels of s-Hcy than those on other AEDs (F=5.89, p=.016). Regression analyses revealed homocysteine, fracture history, length of years on AEDs, ethnicity were predictors of spine T scores. Weight and BMI were predictors of both BMD and DEXA T scores. Use of enzyme-inducing AEDs was a negative predictor of spine BMD and T scores, while phenytoin monotherapy was a positive predictor of spine BMD. Lamotrigine was found to be a negative predictor of spine T score. Ambulatory status, menopause and alcohol consumption were predictors of BMD but not T scores. In this study, persons with epilepsy who take nutritional supplementation have 25% lower s-Hcy levels than those who do not. Supplementation continues to be important in preventative epilepsy care. PMID- 17110135 TI - Evaluation of cartilage repair in the distal femur after autologous chondrocyte transplantation using T2 relaxation time and dGEMRIC. AB - OBJECTIVES: To investigate the potential of combining T2 relaxation time and delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) measurements after autologous chondrocyte transplantation (ACT). MATERIALS AND METHODS: T2 and dGEMRIC maps were measured in the sagittal and coronal directions in 12 patients 10-15 months after ACT surgery. Grafts were assessed for bulk full thickness, superficial and deep tissue T2 and dGEMRIC values, and were compared to control cartilage. RESULTS: All ACT grafts showed filling of the repair area to the level of or above the articular surface. Matrix of the grafts lacked the classical laminar structure and appeared more heterogenous on T2 maps than control cartilage. As compared to control cartilage, ACT grafts showed significantly longer T2 values for bulk tissue as well as for the superficial 50% and deep 50% of tissue except for superficial cartilage in the coronal direction. dGEMRIC assessment in the sagittal and coronal directions did not show a significant difference between bulk, superficial or deep tissue as compared to the control cartilage. Superficial and deep ACT tissue did not differ statistically in terms of their T2 or dGEMRIC values. CONCLUSIONS: These preliminary results suggest that, according to T2 measurements, ACT repair tissue at 10-15 months differs from normal cartilage and probably lacks the preferential collagen arrangement of normal cartilage, while according to dGEMRIC a varying degree of proteoglycan replenishment takes place. Combining these two quantitative magnetic resonance imaging techniques enables a more comprehensive characterization of cartilage repair than before. PMID- 17110136 TI - An animal model of intrauterine growth retardation induced by synthetic thromboxane a(2). AB - OBJECTIVE: Intrauterine growth retardation (IUGR) is an important cause of prenatal and neonatal morbidity, and neurologic abnormalities. Although several animal models of IUGR have been developed for scientific investigation, few models approximate the pathophysiology in human fetal growth failure resulting from pregnancy-induced hypertension and preeclampsia. We developed an animal model of IUGR in which fetal growth restriction was induced by administering a synthetic thromboxane A(2) analogue (STA(2)) to the mother. METHODS: Timed pregnant Sprague-Dawley rats were used in this study. STA(2) was delivered into the peritoneal cavity of the pregnant female at a rate of 20 ng/h from day 13 of pregnancy. The effectiveness of this model was evaluated by monitoring the overall growth of the fetuses and neonates and measuring the weight and biochemical composition of individual organs. RESULTS: Fetuses and neonates from the STA(2) group showed a highly significant weight reduction throughout the observation period from day 19 of gestation to postnatal day 7. Weight reduction near and at term exceeded 10% and became more pronounced during the first week after birth. Fetuses on the 20th gestational day exhibited a pattern of growth retardation characteristic of asymmetrical IUGR in which the weight reduction was prominent in the liver with relative sparing of the brain. However, the decrease in brain weight was more than 10%. The protein, DNA, and RNA contents of the liver were lower in the STA(2) group. The protein content of the forebrain and brainstem also decreased significantly in the STA(2) group compared with the control; however, the DNA content of the forebrain was higher in the STA(2) group. CONCLUSIONS: This animal model may mimic human IUGR more closely than previous models because the growth restriction is induced in a truly chronic manner. PMID- 17110137 TI - Complete ulcer healing as primary endpoint in studies on critical limb ischemia? A critical reappraisal. AB - OBJECTIVES: Although complete ulcer healing is the mandatory primary efficacy criterion in current European guidelines for drug trials in critical limb ischemia (CLI), the appropriateness of this endpoint has been questioned for some time. We carried out a systematic review to assess the value of this endpoint in studies on reconstructive measures, considered to be the standard of care for CLI. METHODS: A computerized literature search (1985-2005) was performed to track down clinical studies on endovascular and surgical interventions by using the search terms CLI and ulcer healing and their synonyms. RESULTS: 1,914 papers on revascularization in CLI were identified. Complete ulcer healing was reported in 17 studies (0.9%). Among these, there were no randomized controlled trials, five prospective cohorts on endovascular procedures, and six retrospective cohorts for endovascular and surgical procedures, respectively. If healing rates or time to ulcer healing were available, they differed greatly between the studies without consistent correlation to types of therapy. CONCLUSIONS: In past and current literature, complete ulcer healing is not a consistently reported criterion for success of revascularization in CLI. Thus, its appropriateness for efficacy assessment of drug studies in CLI patients has to be questioned. PMID- 17110138 TI - HS-48 alone has no enhancement role on the expression of human alpha-globin gene cluster. AB - To investigate the in vivo function of the newly defined DNase I hypersensitive site HS-48 on the whole human alpha-globin gene cluster, the region containing all the other known 5 hypersensitive sites HS-4 to HS-40 was deleted from a 117 kb bacterial artificial chromosome clone bearing the whole human alpha-globin gene cluster. Transgenic mice were generated from this construct. The RNase protection assays showed that with HS-48 left and all the other 5 hypersensitive sites deleted, the expression of human alpha-like globin genes was completely silenced in embryonic, fetal and adult stages in all tissues. This finding indicates that HS-48 alone has no enhancer activity on the expression of human alpha-like globin genes, and that the region of HS-4 to HS-40 already contains all the upstream cis-elements needed for regulating human alpha-like globin genes. PMID- 17110139 TI - Glutamate receptors and pain. AB - Pain is an important survival and protection mechanism for animals. However, chronic/persistent pain may be differentiated from normal physiological pain in that it confers no obvious advantage. An accumulating body of pharmacological, electrophysiological, and behavioral evidence is emerging in support of the notion that glutamate receptors play a crucial role in pain pathways and that modulation of glutamate receptors may have potential for therapeutic utility in several categories of persistent pain, including neuropathic pain resulting from injury and/or disease of central (e.g., spinal cord injury) or peripheral nerves (e.g., diabetic neuropathy, radiculopathy) and inflammatory or joint-related pain (e.g., rheumatoid arthritis, osteoarthritis). This review focuses on the role of glutamate receptors, including both ionotropic (AMPA, NMDA and kainate) and metabotropic (mGlu1-8) receptors in persistent pain states with particular emphasis on their expression patterns in nociceptive pathways and their potential as targets for pharmacological intervention strategies. PMID- 17110140 TI - The ADHD and sleep conundrum redux: moving forward. PMID- 17110141 TI - Effects of exercise and oral antioxidant supplementation enriched in (n-3) fatty acids on blood oxidant markers and erythrocyte membrane fluidity in horses. AB - The aim of this study was to investigate in a placebo-controlled field study the effect of a (n-3)-vitamin supplementation on erythrocyte membrane fluidity (EMF), oxidant/antioxidant markers and plasmatic omega3/omega6 fatty acid ratio (FAR) in 12 eventing horses. Venous blood was sampled at rest before (PRE) and after (POST) a three week treatment period with either the supplement (group S, n=6) or a placebo (group P, n=6) as well as after 15min (POST E15') and 24h (POST E24h) after a standardised exercise test. The following markers were analysed: EMF, plasma antioxidant capacity of water and lipid soluble components, ascorbic acid, uric acid (UA), glutathione (reduced: GSH, oxidised: GSSG), vitamin E (Vit E), beta-carotene, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity, selenium, copper (Cu), zinc (Zn), oxidised proteins (Protox), lipid peroxides (Pool) and FAR. EMF did not differ between group S and P after treatment, but GPx remained unchanged in group S whereas it decreased in group P and plasma Cu/Zn ratio remained unchanged whereas it increased in group P. FAR were significantly increased in group S. Exercise induced a significant decrease of EMF (POST vs. E24h) in both groups, but which was significantly lower at E15' in group S than in group P. Exercise induced a significant increase of UA and ACW (POST vs. E15') and Protox (POST vs. E24h) in both groups. An exercise-related decrease in GSH and Pool (POST vs. E15') was found in group P, whereas Vit E and FAR (POST vs. E24h) significantly decreased in both groups. The study showed that exercise induced a decrease in EMF in horses associated with changes of blood oxidative balance. The (omega-3)-vitamin supplementation tested improved the oxidative balance poorly but delayed the exercise-induced decrease of EMF and increased the FAR. PMID- 17110142 TI - Effects of housing, parturition and diet change on the biochemistry and biomechanics of the support structures of the hoof of dairy heifers. AB - This study investigated effects of housing, diet and parturition on the biochemistry, biomechanics and pathology of feet of maiden, pregnant and lactating dairy heifers. Strength/laxity, laminar morphology, connective tissue (CT) biochemistry and sole lesions were assessed. Although no animals became clinically lame, severity of sole lesions was significantly greater in heifers housed in cubicles vs. straw yards, and in lactating/pregnant heifers vs. maidens. These effects were additive. Cubicle housing and parturition each increased CT metabolism (and were additive), and altered CT composition. Similarly, both impaired the biomechanical resilience of the hoof. There were no effects for diet. The results indicate that parturition/lactation causes non inflammatory changes in CT that impair resilience of the feet to external stresses associated with poor housing. This "parturition effect" appears to be unrelated to change in diet and relatively brief, unless exacerbated by additional stresses of housing. Thus heifer lameness may be significantly reduced through short-term, low-cost improvements in husbandry around the time of calving. PMID- 17110143 TI - Localisation and modulation of prostanoid receptors EP1 and EP4 in the rat chronic constriction injury model of neuropathic pain. AB - Immunohistochemistry was used to examine the expression of prostaglandin E(2) receptors EP1 and EP4 in sciatic nerves from the rat chronic constriction injury (CCI) model of neuropathic pain. At 21 days post-surgery the CCI rats had developed mechanical hyperalgesia on the operated side, and quantitative image analysis showed a highly significant doubling of the area occupied by EP1- and EP4-positive pixels in sections from CCI nerves when compared to sham-operated controls. Co-localisation studies with the marker ED1 revealed that 73% of the EP1-positive cells and 54% of the EP4-positive cells in the injured nerves represented infiltrating macrophages. Cells negative for ED1 and positive for either EP1 or EP4 were characterised as Schwann cells from their morphology and expression of myelin basic protein and S100 antigens. Similar EP1- and EP4 positive Schwann cell profiles were observed in sections of uninjured control nerves. Low levels of EP receptor expression were found in neurofilament immunostained axons, but no consistent differences were observed in the levels of axonal EP staining between CCI and control tissue. These data provide further evidence of the importance of prostaglandins in the pathogenesis of neuropathic pain, and suggest that not only infiltrating macrophages but also Schwann cells may be involved in the modulation of these mediators in response to nerve injury. PMID- 17110144 TI - Diffraction-like phenomena in a periodic magnetization distribution at 1.5 T using the distant dipolar field (DDF). AB - In the CRAZED experiment (COSY revamped by asymmetric Z-gradient echo detection, Warren et al.), a spatially anisotropic magnetization distribution is created by application of a magnetic field gradient (strength G, duration tau) which in turn generates a response called the distant dipolar field (DDF). The DDF is a source of intermolecular multiple-quantum coherences (iMQC) which contain information on the distance d=pi/(gammaGtau) between pairs of dipolar-coupled spins. Diffraction like phenomena may result for periodically structured samples. In this study, we report the observation of diffraction owing to the DDF at 1.5 T using a clinical whole-body tomograph. Based on the semi-classical treatment of the problem by Robyr and Bowtell, diffraction conditions were obtained for a CRAZED-type pulse sequence that selects iMQC of order N. The predicted distinct difference in N=2 and N not equal2 coherences, i.e., a dominant continuous course as a function of tau (N=2) and prominent diffraction peaks otherwise, could be verified in CRAZED experiments in a periodically structured sample selecting coherence orders N=2 and N=3. The diffractive signal component contains information on the geometric structure of the sample. Applications of this technique may permit the detection of changes in composition and geometry of periodic structures. PMID- 17110145 TI - Atom-based stochastic and non-stochastic 3D-chiral bilinear indices and their applications to central chirality codification. AB - Non-stochastic and stochastic 2D bilinear indices have been generalized to codify chemical structure information for chiral drugs, making use of a trigonometric 3D chirality correction factor. In order to evaluate the effectiveness of this novel approach in drug design we have modeled the angiotensin-converting enzyme inhibitory activity of perindoprilate's sigma-stereoisomers combinatorial library. Two linear discriminant analysis models, using non-stochastic and stochastic linear indices, were obtained. The models had shown an accuracy of 95.65% for the training set and 100% for the external prediction set. Next the prediction of the sigma-receptor antagonists of chiral 3-(3 hydroxyphenyl)piperidines by multiple linear regression analysis was carried out. Two statistically significant QSAR models were obtained when non-stochastic (R(2)=0.953 and s=0.238) and stochastic (R(2)=0.961 and s=0.219) 3D-chiral bilinear indices were used. These models showed adequate predictive power (assessed by the leave-one-out cross-validation experiment) yielding values of q(2)=0.935 (s(cv)=0.259) and q(2)=0.946 (s(cv)=0.235), respectively. Finally, the prediction of the corticosteroid-binding globulin binding affinity of steroids set was performed. The obtained results are rather similar to most of the 3D-QSAR approaches reported so far. The validation of this method was achieved by comparison with previous reports applied to the same data set. The non-stochastic and stochastic 3D-chiral linear indices appear to provide a very interesting alternative to other more common 3D-QSAR descriptors. PMID- 17110146 TI - Molecular modeling of purinergic receptor P2Y12 and interaction with its antagonists. AB - Purinergic receptors are a class of cell surface receptors for purines that prefer ATP or ADP over adenosine. The surface receptors for extracellular nucleotides are called P2 receptors. They are activated by both pyrimidine and purine nucleotides. ADP initiates platelet aggregation by 'simultaneous activation of two G protein-coupled receptors, P2Y1 and P2Y12. P2Y12 has been shown to be the target of the thienopyridine drugs, ticlopidine and clopidogrel. Here, the active sites of P2Y12 for ATP as well as ADP are predicted by bioinformatics and molecular modeling. First, the three-dimensional (3D) structure of P2Y12 was constructed by InsightII/Homology module using the corresponding bovine rhodopsin (PDB code: 1HZX) as the template. Then the primary structures were optimized by energy minimization that has been successfully accepted by the Protein Data Bank (PDB code: 1VZ1). Second, a simple scoring matrix was built up based on the analysis of 13 known ATP-binding proteins. And the most probable active sites of P2Y12 were predicted using the scoring matrix, which include three distant areas: "head area" (LGTGPLRTFV, 87-96), "middle area" (VGLITNGLAM, 38-47, and LGAKILSVVI, 139-148), and "bottom area" (RTRGVGKVPR, 222 231). Subsequently the structural model of P2Y12 was docked with ATP/ADP in comparison with P2Y1 (PDB code 1ddd). As a comparison, we docked its antagonists, such as ticlopidine and clopidogrel, to the most probable sites and calculated their intermolecular energy. Our results imply that P2Y12 has the potential to be inhibited by ADP/ATP analogs, and it suggests that P2Y12 acts as a target of new drugs that inhibit platelet aggregation. PMID- 17110147 TI - Differences in the physiological responses to temperature among stonechats from three populations reared in a common environment. AB - The physiological response to variation in air temperature (T(a)) can provide insights into how animals are adapted to different environments. I measured metabolic rate, total evaporative water loss (TEWL) and body temperature (T(b)) as a function of T(a) in stonechats from equatorial Kenya, temperate central Europe and continental Kazakhstan, environments where stonechats have evolved different life histories. All birds were raised and kept under identical captive conditions to highlight genetically based differences and to exclude phenotypic plasticity as explanatory factor. The slope relating metabolic rate to T(a) was steepest in Kazakh stonechats and lowest for birds from Kenya, indicating that, counterintuitively, the tropical stonechats were best insulated. Taking into account variation in T(b) in response to T(a), the lower critical temperature for the three populations fell between 32.0 and 34.9 degrees C, values higher than previously assumed. Whole organism BMR did not differ among populations, but because body mass was significantly higher in the Kenyan stonechats, their mass specific BMR was lower compared with conspecifics from higher latitude. Whole organism or mass-specific TEWL did not differ among populations. Possibly, Kenyan birds are better insulated to compensate for their limited capacity to elevate metabolic rate. PMID- 17110148 TI - [Epidermoid carcinomas of anal canal treated with radiation therapy and concomitant chemotherapy (5-fluorouracil and cisplatin)]. AB - PURPOSE: To evaluate our results after radiation therapy and concomitant chemotherapy in terms of local control, survival and toxicity in patients with anal cancer. METHODS AND PATIENTS: Between November 1990 and January 2002, 60 patients (pts) were treated with radiation therapy and concomitant chemotherapy. The T-stage according to the 2001 UICC classification were: 2 T1, 26 T2, 25 T3, and 7 T4. There were 20 pts with nodal involvement at presentation. The treatment started with external beam RT (median dose: 45 Gy) and concomitant chemotherapy using 5-fluorouracil and cisplatin during the first week and the fifth week of external beam RT (EBRT). After a rest period of 4 to 6 weeks, a boost of 20 Gy was delivered by EBRT in 58 pts and by interstitial (192)Ir brachytherapy in 2 pts. Mean follow-up were 78.5 months. RESULTS: At the end of RT with concomitant chemotherapy local tumor clinical complete response rate was 83%. Out of 10 non responders or local progression, 5 (50%) were salvaged with abdominoperineal resection (APR). Out of 5 local tumor relapses, 3 were salvaged with APR. The overall local tumor control (LC) rate with or without salvage local treatment were 88%. LC rate with a good anal function scoring (score 0 and 1) was 70%. Among 43 pts who preserved their anus, 98% had a good anal function scoring. The 5-year disease-free survival was 75%. After multivariate analysis, 2 independent predicting factors significantly influenced the disease-free survival: HIV positive pts (negative vs positive, P=0.032) and clinical tumor response after the first course of radiotherapy (<50% vs >or=50%, P=0.00032). Acute grade 2 or 3 toxicities were low: haematological toxicity in 4 pts and intestinal complication corresponding to diarrhea in 10 pts. Late severe complication was observed in 3 pts: 2 pts with painful necrosis of the anus requiring colostomy and 1 pt with grade 3 rectal bleeding. CONCLUSION: We confirm the good results with RT and concomitant chemotherapy. The clinical tumor response after the first course of RT and concomitant chemotherapy is probably the most important predictive factor on the disease-free survival. For patients with T3 or T4 lesion and tumor regression (7)F(4)) and red (660 nm: (5)D(4)-->(7)F(2)) up-conversion (UC) luminescence originating from Tb3+ is observed strongly, because of the quadratic dependences of emission intensities on the excitation power. Especially, the UC luminescence was intensified violently with the energy transfer from the Tm3+ ions involves in the Tb3+ excitation. To the Tb3+/Tm3+/Yb3+ triply doped glass system, a novel up-conversion mechanism is proposed as follows: the energy of (3)G(4) level (Tm3+) was transferred to (5)D(4) (Tb(3+)) and the 477-nm UC luminescence of Tm3+ was nearly quenched. PMID- 17110159 TI - Determination of trace amounts of thiocyanate by a new kinetic procedure based on an induction period. AB - A new, simple and sensitive kinetic spectrophotometric method with no need for removing of interfering substances is proposed for the determination of thiocyanate ion in biological and water samples. The procedure is based on the inhibiting effect of thiocyanate on the sodium periodate-potassium bromide-meta cresol purple (MCP) system in acidic media. The induction period of the reaction is proportional to the SCN- concentration. The decolorization of meta cresol purple by the reaction products was used to monitor the reaction spectrophotometrically at 525 nm. Under optimum conditions, thiocyanate can be determined in the range of 0.02-0.8 microg ml(-1) with a 3sigma detection limit of 5 ng ml(-1). The relative standard deviations for 10 replicate determinations of 0.060, 0.10 and 0.50 microg ml(-1) thiocyanate are 3.7, 2.4 and 1.0%, respectively. This method has been successfully used to the determination of thiocyanate content in smokers and non-smokers saliva and spiked water sample. PMID- 17110160 TI - FT-IR and NMR investigation of 1-phenylpiperazine: a combined experimental and theoretical study. AB - FT-IR and (1)H, (13)C, DEPT, COSY, NOESY, HETCOR, INADEQUATE NMR spectra of 1 phenylpiperazine (pp) have been reported for the first time except for its (1)H NMR spectrum. The vibrational frequencies and (1)H, (13)C NMR chemical shifts of pp (C(10)H(14)N(2)) have been calculated by means of the Hartree-Fock (HF) and Becke-Lee-Yang-Parr (BLYP) or Becke-3-Lee-Yang-Parr (B3LYP) density functional methods with 6-31G(d) and 6-31G(d,p) basis sets, respectively. Comparison between the experimental and the theoretical results indicates that density functional B3LYP method is superior to the scaled HF and BLYP approach for predicting vibrational frequencies and NMR properties. PMID- 17110161 TI - Towards a hybrid method to categorize interruptions and activities in healthcare. AB - OBJECTIVE: Interruptions are known to have a negative impact on activity performance. Understanding how an interruption contributes to human error is limited because there is not a standard method for analyzing and classifying interruptions. Qualitative data are typically analyzed by either a deductive or an inductive method. Both methods have limitations. In this paper, a hybrid method was developed that integrates deductive and inductive methods for the categorization of activities and interruptions recorded during an ethnographic study of physicians and registered nurses in a Level One Trauma Center. Understanding the effects of interruptions is important for designing and evaluating informatics tools in particular as well as improving healthcare quality and patient safety in general. METHOD: The hybrid method was developed using a deductive a priori classification framework with the provision of adding new categories discovered inductively in the data. The inductive process utilized line-by-line coding and constant comparison as stated in Grounded Theory. RESULTS: The categories of activities and interruptions were organized into a three-tiered hierarchy of activity. Validity and reliability of the categories were tested by categorizing a medical error case external to the study. No new categories of interruptions were identified during analysis of the medical error case. CONCLUSIONS: Findings from this study provide evidence that the hybrid model of categorization is more complete than either a deductive or an inductive method alone. The hybrid method developed in this study provides the methodical support for understanding, analyzing, and managing interruptions and workflow. PMID- 17110162 TI - Evaluation of NucliSens EasyQ HIV-1 assay for quantification of HIV-1 subtypes prevalent in South-east Asia. AB - BACKGROUND: Monitoring anti-retroviral therapy requires that viral load assays for human immunodeficiency virus type 1 (HIV-1) be applicable to diverse HIV-1 subtypes. OBJECTIVES: To evaluate NucliSens EasyQ HIV-1 assay for quantitation of common HIV-1 subtypes prevalent in South-east Asia. STUDY DESIGN: One hundred and nineteen plasma samples collected in Hong Kong and Cambodia were used to compare the performance of NucliSens EasyQ HIV-1 and COBAS Amplicor HIV-1 Monitor version 1.5 assays. Viral RNA extracted from the NucliSens MiniMAG was also used for HIV 1 subtyping. RESULTS: Performance of NucliSens EasyQ correlated well with COBAS Amplicor (r=0.777, p<0.001) and the small mean difference (0.0462log(10)IU/mL) obtained in the Bland and Altman model indicated good agreement between two assays. The NucliSens EasyQ assay demonstrated a 95% sensitivity at 500IU/mL and 100% specificity. Reproducibility of this assay was within log(10)2-4IU/mL and had a coefficient of variation between 2.3% and 10.4%. Among the 109 specimens included in the analysis, HIV-1 subtyping identified 64 CRF01_AE, 38 subtype B, 3 subtype C, 3 CRF07_BC and 1 subtype G viruses. CONCLUSIONS: Performance of NucliSens EasyQ was comparable to COBAS Amplicor for HIV-1 viral load monitoring. RNA extracts from NucliSens MiniMAG could be used for HIV-1 viral load monitoring, subtyping and drug resistance mutations detection. Our findings highlight the versatility of both NucliSens EasyQ and COBAS Amplicor in monitoring prevalent subtypes and rare circulating recombinant forms (CRFs) in the South-east Asia region. PMID- 17110163 TI - Effects of FXR in foam-cell formation and atherosclerosis development. AB - Farnesoid X receptor (FXR), a bile-acid-activated member of the nuclear receptor superfamily, is essential in regulating bile-acid, cholesterol, and triglyceride homeostasis. Disruption of the FXR gene in mice results in a proatherosclerotic lipid profile with increased serum cholesterols and triglycerides. However, the role of FXR in foam-cell formation and atherosclerosis development remains unclear. The current study showed that the peritoneal macrophages isolated from FXR-null mice took up less oxidized LDL-cholesterol (oxLDL-C), which was accompanied by a marked reduction in CD36 expression in these cells. This result appears to be FXR-independent, as FXR was not detected in the peritoneal macrophages. To assess to what extent FXR modulates atherosclerosis development, FXR/ApoE double-null mice were generated. Female mice were used for atherosclerosis analysis. Compared to ApoE-null mice, the FXR/ApoE double-null mice were found to have less atherosclerotic lesion area in the aorta, despite a further increase in the serum cholesterols and triglycerides. Our results indicate that disruption of the FXR gene could attenuate atherosclerosis development, most likely resulting from reduced oxLDL-C uptake by macrophages. Our study cautions the use of serum lipid levels as a surrogate marker to determine the efficiency of FXR modulators in treating hyperlipidemia. PMID- 17110164 TI - Mutagenesis of rat acyl-CoA synthetase 4 indicates amino acids that contribute to fatty acid binding. AB - Although each of the five mammalian long-chain acyl-CoA synthetases (ACSL) can bind saturated and unsaturated fatty acids ranging from 12 to 22 carbons, ACSL4 prefers longer chain polyunsaturated fatty acids. In order to gain a better understanding of ACSL4 fatty acid binding, we based a mutagenesis approach on sequence alignments related to ttLC-FACS crystallized from Thermus thermophilus HB8. Four residues selected for mutagenesis corresponded to residues in ttLC-FACS that comprise the fatty acid binding pocket; the fifth residue aligned with a region thought to be involved in fatty acid selectivity of the Escherichia coli acyl-CoA synthetase, FadD. Changing an amino acid at the entry of the putative fatty acid binding pocket, G401L, resulted in an inactive enzyme. Mutating a residue near the pocket entry, L399M, did not significantly alter enzyme activity, but mutating a residue at the hydrophobic terminus of the pocket, S291Y, altered ACSL4's preference for 20:5 and 22:6 and increased its apparent K(m) for ATP. Mutating a site in a region previously identified as important for fatty acid binding also altered activation of 20:4 and 20:5. These studies suggested that the preference of ACSL4 for long-chain polyunsaturated fatty acids can be modified by altering specific amino acid residues. PMID- 17110165 TI - Genomic structure, alternative maturation and tissue expression of the human BBOX1 gene. AB - Gamma-butyrobetaine hydroxylase (BBOX1) is the enzyme responsible for the biosynthesis of l-carnitine, a key molecule of fatty acid metabolism. This cytosolic dimeric protein belongs to the dioxygenase family. In human, enzyme activity has been detected in kidney, liver and brain. The human gene encoding gamma-butyrobetaine hydroxylase is located on chromosome 11. Although the protein structure and activity have been extensively described, little information is available concerning BBOX1 structure and expression. In this study, the organization of the human gene was determined. The structure and functions of the 5'- and 3'-untranslated regions of the human BBOX1 mRNA were characterized in kidney, liver and brain. Our experiments revealed that the transcription initiation of the human BBOX1 gene might occur at 3 different exons, and that the expression level of each type of transcript is organ-specific. We showed that the use of 3 different promoters is responsible for the 5'-end heterogeneity. Investigations on BBOX1 mRNA maturation highlighted an alternative polyadenylation mechanism that generates two 3'-untranslated regions differing by their length. This alternative polyadenylation exhibited a tissue specificity. PMID- 17110166 TI - Feature-similarity protein classifier as a ligand engineering tool. AB - Kinases have been often targeted in drug therapy aimed at blocking signaling pathways. However, the conservation of protein structure across homologs often leads to uncontrolled cross-reactivity. On the other hand, sticky packing defects in proteins are typically not conserved across homologs, making them ligand anchoring sites potentially important to enhance selectivity. Thus, we introduce a hierarchical clustering of PDB-reported kinases according to packing differences. This kinome partitioning is highly correlated with proximity relations arising from the pharmacological profiling of kinases. A variable packing sensitivity is observed for individual drugs, with highly promiscuous ligands being the most insensitive to packing differences. Our classifier enables a strategy to design selective inhibitors. PMID- 17110167 TI - Blood purification therapy for sepsis. AB - Accumulating evidences of underlining pathogenesis of sepsis have contributed to the therapeutic strategy for sepsis. Not only endotoxin and cytokine, but also signal transduction through Toll-like receptors could be a strategic target for the management of sepsis. Blood purification therapy including polymyxin B immobilized hemoperfusion cartridge and continuous hemodiafiltration has shown the beneficial effect on patients with sepsis in Japan. Although they were initially designed to remove endotoxin and cytokines respectively, they might eliminate unexpected mediators responsible for sepsis. Further elucidation of mechanism and randomized controlled studies are needed to establish the role of blood purification therapy in sepsis. PMID- 17110168 TI - Pathogen-reduction systems for blood components: the current position and future trends. AB - The current multi-layered interventional approaches to blood safety have dramatically reduced the risk of viral contamination of blood components. Nowadays most of the residual transfusion transmitted infections (TTI) occur as the result of the interval between the time the donor is infected and the moment at which tests are capable of detecting the agent, the so called "window period" which has been considerably reduced by the increased sensitivity of nucleic acid testing (NAT). However, the residual risk of bacterial contamination and the unexpected appearance of some other emerging pathogens, almost every five years, are still of major concern to the public, politicians, regulatory agencies and place immense pressures on the organisations responsible for the provision of safe blood and its components. In view of these bleak scenarios, the use of human blood as a raw biological source is inherently unsafe, and screening/testing alone cannot exclude all the potential human pathogens; hence we need to put in place some sort of safer alternatives and/or additional preventative safety measures. Recently, several substitutes (alternatives) to virtual blood components have been developed and tried. Moreover, various mechanical methods such as cell washing and leukofiltration have been implemented as additional preventative safety measures but with limited success in abrogating the risk of transfusion transmitted cell-associated agents. The most promising approaches, so far, are methods that target pathogen nucleic acids (Methylene blue; Psolaren and Riboflavin UV light treatment). These procedures have undergone considerable in vitro studies to ensure their extremely high safety margins in terms of toxicity to the cells or to the recipients. In essence, while the technology of targeting nucleic acid to stop viral proliferation is common to the above three strategies, in practice these procedures differ in terms of operational, physicochemical and biological characteristics; including the potential impacts of their metabolites and photo-adducts; their effects on the spectrum of pathogens affected and the log reductions in culture infective studies. Accordingly, any strategy that involves addition of an extraneous agent or physicochemical manipulation of blood must balance the benefits of pathogen reduction against the loss or alteration to the cells and plasma functional integrity, short and long term toxicity to the cells and to the recipients, as well as the risk to the personnel involved and the community at large. Moreover, it must be noted that each method will have a different profile of adverse reactions and may differ in terms of the risk to particularly vulnerable groups of patients, requiring in depth clinical trials, while taking into consideration the cost benefit of the final process. Newer diagnostic procedures must be in place to establish the storage stability of products that have undergone pathogen inactivation, in particular tests reflecting the release of platelet-derived cytokines, cellular apoptosis or microvesiculation and their role in immunosupressiveness. This overview aims to provide an update on the continual improvements in blood component safety, in particular using methods that target pathogen nucleic acid. Emphasis is placed on methylene blue light treatment (MBLT) and Intercept or Mirasol PRT systems for platelets and plasma. The status of pathogen reduction of whole blood and red cells is also highlighted, though the progress in this area has been virtually stopped after the finding of antibody development in the clinical trial. PMID- 17110170 TI - Religion, infertility and assisted reproductive technology. AB - This chapter describes religion in general before discussing the centrality of its concern for family formation. In light of this, the impact of infertility on religious people is considered. Recognizing religion's cautiously positive attitude towards assisted reproductive technology (ART) as a potential ally in the project of family formation and the relief of infertility, two areas that have caused concern for the religions are discussed: perceived threats to marriage and the sanctity of the human embryo. Throughout the chapter, illustrations are drawn from particular religions, including Christianity, Judaism, Islam, Hinduism and Buddhism. There are striking similarities in their concerns and in the range of their responses to ART. Ways in which medical personnel should take into account the religious dimensions of the experience of infertility in their care for patients are suggested. PMID- 17110171 TI - Thyroid hormone status of Atlantic croaker exposed to Aroclor 1254 and selected PCB congeners. AB - Atlantic croaker (Micropogonias undulatus) were exposed to the polychlorinated biphenyl (PCB) mixture (Aroclor 1254) or one of three individual congeners (planar PCB 77 or ortho-substituted PCB 47 and PCB 153) in the diet for 30 days to investigate the effects of PCBs on circulating thyroid hormones, thyroxine (T4) and triiodothyronine (T3). Aroclor 1254 (0.2 and 1.0 mg/kg body mass/day) decreased plasma T3 levels consistently, but the effects on T4 levels were inconsistent from year to year. Exposure to PCB 153 (0.1 and 1.0 mg/kg body mass/day) significantly lowered both T4 and T3, while PCB 47 at the same doses had no effect on thyroid hormone levels. The lower doses of PCB 77 (0.004, 0.01 and 0.02 mg/kg body mass/day) had no effect on T4 or T3, whereas the highest dose (0.1 mg/kg body mass/day) increased T4 levels significantly. The results of the present study demonstrate that exposure to PCBs at environmentally realistic concentrations can have profound effects on the thyroid status of Atlantic croaker. The ortho-substituted PCB 153 appears to contribute at least partially to the deleterious effects of Aroclor 1254 on thyroid status, whereas the planar PCB 77 at concentrations present in the mixture is unlikely to alter thyroid hormone levels. PMID- 17110172 TI - Cluster-crossover design: a method for limiting clusters level effect in community-intervention studies. AB - The cluster-crossover design can be used for clinical trials comparing two or more interventions in a naturally formed study population, i.e. a cluster. This design differs from that of a crossover study, in that the treatment sequence is allocated at the cluster level. A cluster-crossover study can thus be considered as a cluster-randomized controlled study with additional periodic cluster randomization(s) or treatment permutation(s) during the study. The data must be analyzed with hierarchical models with random effects in order to allow for different outcome probabilities in each period, cluster and cluster-period. Original data from two published field studies of hospital infection control based on this design are used here to illustrate the impact of different statistical models on the interpretation of the results. PMID- 17110173 TI - Expression of the forkhead transcription factor FoxN4 in progenitor cells in the developing Xenopus laevis retina and brain. AB - Forkhead proteins are involved in gene regulation in a large variety of developmental situations. Several forkhead gene products are expressed in the developing eye and brain. Here we characterize the expression of FoxN4 during Xenopus development. We report that FoxN4 is expressed in the eye from the earliest stages of specification through retinal maturation. FoxN4 is also expressed in the pallium, optic tectum, isthmus, reticular formation, and in cells lining the ventricle of the tadpole brain. PMID- 17110174 TI - Genetic characterization of group A rotavirus strains circulating among children with acute gastroenteritis in Japan in 2004-2005. AB - A total of 752 fecal specimens collected from July 2004 to June 2005 from children with acute gastroenteritis in four localities in Japan (Maizuru, Tokyo, Sapporo, and Osaka) were screened for group A rotavirus by RT-PCR. It was found that 82 (10.9%) specimens were positive for group A rotavirus. The G-(VP7 genotypes) and P-(VP4 genotypes) types were further investigated. The P-types of 18 rotavirus strains, which could not be typed by RT-PCR, were determined by sequencing analysis. Of these, 94% (17/18) were P[8] with multiple point mutations at the VP4 primer-binding site. Another sample turned out to be a rare genotype P[9], which was closely related to feline rotavirus. The predominant genotype was G1P[8] (46.4%), followed by G3P[8] (32.9%) and G2P[4] (12.2%). A number of unusual combinations including, G1P[4] (1.2%), G2P[8] (1.2%), G3P[9] (1.2%), G1G3P[8] (1.2%), and G2G3P[8] (3.7%), were also detected. A new nomenclature of P[8] was proposed, in which worldwide rotavirus P[8] strains were classified into four sub-lineages, namely IA, IB, IIA, and IIB. A wide range of amino acid substitutions (up to 22) specific for P[8] lineages and sub-lineages were also identified. Interestingly, only short amino acid motifs located at positions 32-35, 121-135, and 195-236 of VP4 correctly defined the phylogenetic P[8] lineages and sub-lineages. Of note, at least two distinct clusters of rotavirus P[8] were co-circulating in the Japanese pediatric population studied. PMID- 17110175 TI - Mitochondrial upstream promoter sequences modulate in vivo the transcription of a gene in yeast mitochondria. AB - An in vivo study of the importance of the length and/or structures of sequences upstream of a mitochondrial promoter was undertaken in Saccharomyces cerevisiae. Short tandem mtDNA repeats were introduced upstream of the COX2 gene. Our data show that its expression is modulated by the sequence located over 200 bp upstream of the promoter. A deletion decreases the level of transcripts to about 50%. The initial level can be recovered by a fill-in AT-rich sequence or partially by the presence of a long repeat tract; on the contrary, a smaller number of copies tends to intensify the effect of the deletion. These results show that the length and base composition upstream of mitochondrial promoter are involved in vivo in the modulation of the gene expression. PMID- 17110176 TI - Arginine, citrulline and nitrate concentrations in the cerebrospinal fluid from patients with acute hydrocephalus. AB - Citrulline and nitric oxide (NO) are synthesized by NO synthase (NOS) in a 1:1 stoichiometry. In this study, we determined by HPLC arginine and citrulline concentrations by fluorescence detection and nitrate levels by UV absorbance detection in the cerebrospinal fluid (CSF) from patients with acute hydrocephalus that underwent ventricular drainage. We found increased citrulline concentration (50.6+/-17.2 versus 20.9+/-2.0 microM) and decreased arginine/citrulline molar ratio (0.42+/-0.11 versus 1.12+/-0.16) in hydrocephalus patients, while arginine and nitrate concentrations and citrulline/nitrate molar ratio remained with little change. Citrulline has been determined as a marker of NOS activity in some studies, but it remains to be determined the extent at which this statement holds true, since other biochemical pathways also regulate the concentration of this amino acid. Our results suggest that citrulline is primarily synthesized from NOS in acute hydrocephalus. The evaluation of sample deproteinization by addition of methanol for the analysis of amino acids in CSF is also reported. PMID- 17110177 TI - Radiochromatographic assay of metabolites of the oostatic peptide labeled in different positions of the peptide chain. AB - Reversed-phase high-performance liquid radio-chromatography (radio-HPLC) was set up to detect the time course of labeled degradation product formation of the pentapeptide H-Tyr-Asp-Pro-Ala-Pro-OH (5P), which has oostatic effects in different insect species. The detection limit of the system was in the range of 80-150 Bq. To follow formation of the degradation products, three amino acid residues in 5P were independently tritiated: Tyr1, Pro3 and Pro5. Each of the three tritiated peptides was analyzed after incubation with fresh hemolymph or ovaries of Neobellieria bullata. In the incubation mixture, free terminal amino acids and shortened sequences of 5P were identified. A metabolite of tyrosine represented the only exception; it was finally identified as water using degradation of [3H]Tyr by tyrosinase. Metabolic degradation of [3H]Tyr-5P was found to be considerably quicker than that of H-[3H]Tyr-Asp-Pro-Ala-OH (4P). The degradation of 5P was considerably slower in ovaries in comparison to hemolymph. PMID- 17110178 TI - Separation, characterization and identification of boar seminal plasma proteins. AB - Methods used for the isolation, separation and characterization of boar seminal plasma proteins are discussed, as well as techniques applied to study their binding properties. Attention is paid to interactions of these proteins with different types of saccharides and glycoconjugates, with membrane phospholipids, and to interactions between proteins. Boar seminal plasma contains different types of proteins: spermadhesins of the AQN and AWN families; DQH and PSP proteins belong to the most abundant. Some of these proteins are bound to the sperm surface during ejaculation and thus protein-coating layers of sperm are formed. Sperms coated with proteins participate in different types of interactions occurring in the course of the reproduction process, e.g. formation of the oviductal sperm reservoir, sperm capacitation, oocyte recognition and sperm binding to the oocyte. PMID- 17110179 TI - A simple and rapid HPLC/UV method for the simultaneous quantification of theophylline and etofylline in human plasma. AB - A simple, sensitive and selective high performance liquid chromatography (HPLC) method with ultraviolet detection (272 nm) was developed and validated for the simultaneous quantification of theophylline and etofylline in human plasma. Following rapid sample preparation, the analytes and internal standard (hydrochlorothiazide) were separated using an isocratic mobile phase on a reverse phase C18 column. The lower limit of quantification was 100 ng/mL for both theophylline and etofylline with a relative standard deviation of less than 6%. A linear dynamic range of 100-10,000 ng/mL for both theophylline and etofylline was established. This HPLC method was validated with between-batch precision of 2.2 6.0 and 1.4-3.7% for theophylline and etofylline, respectively. The between-batch accuracy was 94.3-98.0 and 95.4-98.2%, respectively. Stability of theophylline and etofylline in plasma was excellent, with no evidence of degradation during sample processing (autosampler) and 30 days storage in a freezer. This validated method is simple and rugged enough to be used in pharmacokinetic studies. PMID- 17110180 TI - Hydrophobic modifications of Ras proteins by isoprenoid groups and fatty acids- More than just membrane anchoring. AB - During the last years, post-translational modification of peripheral membrane proteins with hydrophobic side groups has been attributed to a couple of additional functions than just simple anchoring into lipid bilayers. In particular isoprenylation and N- and S-acylation did quicken interest in terms of specific recognition elements for protein-protein interactions and as hydrophobic switches that allow for temporal regulated association with distinct target structures. Furthermore new insights into the heterogeneity of natural membranes have connected the physical properties of e.g. farnesyl or palmitoyl side chains with a preference for such sub-compartments as lipid rafts or caveolae. In this review the impact of the two frequently realized modifications by isoprenylation and S-acylation on the process of cellular signal transduction is exemplified with proteins of the Ras and Rab family of small GTP-binding proteins. PMID- 17110183 TI - Dynamic green fluorescent protein sensors for high-content analysis of the cell cycle. AB - We have developed two dynamic sensors that report cell cycle position in living mammalian cells. The sensors use well-characterized components from proteins that are spatially and temporally regulated through the cell cycle. Coupling of these components to Enhanced Green Fluorescent Protein (EGFP) has been used to engineer fusion proteins that report G1/S and G2/M transitions during the cell cycle without perturbing cell cycle progression. Expression of these sensors in stable cell lines allows high content analysis of the effects of drugs and gene knockdown on the cell cycle using automated image analysis to determine cell cycle position and to abstract correlative data from multiplexed sensors and morphological analysis. PMID- 17110184 TI - High-content fluorescence-based screening for epigenetic modulators. AB - Epigenetic processes have gained a great amount of attention in recent years, particularly due to the influence they exert on gene transcription. Several human diseases, including cancer, have been linked to aberrant epigenetic pathways. Consequently, the cellular enzymes that mediate epigenetic events, including histone deacetylases and DNA methyltransferases, have become prime molecular targets for therapeutic intervention. The effective and specific chemical inhibition of these activities is a top priority in cancer research and appears to have therapeutic potential. This chapter describes the development of mammalian cell-based fluorescent assays to screen for epigenetic modulators using an innovative combination of approaches. Detailed protocols for the use of the assays in drug screens, as well as for the initial characterization of hits, are provided. Furthermore, options for evaluating the mechanism of action of these compounds are presented and principles to govern the choice of hit compounds for the development of leads are discussed. PMID- 17110185 TI - Development of assays for nuclear receptor modulators using fluorescently tagged proteins. AB - This chapter describes a method for designing cell-based assays to screen for nuclear receptor modulators. The basic strategy consists in following the movement of the receptors from the cytoplasm into the nucleus in response to ligand binding or analogous activating events. The receptors are tagged with green fluorescent protein for automated, fluorescent detection. In the case of constitutively nuclear receptors, they are engineered for cytoplasmic retention in the absence of an activating signal by fusing them to specific regions of the glucocorticoid receptor, which is found predominantly in the cytoplasm of cultured cells. The resulting chimeras respond to ligands or receptor modulators by translocating into the nucleus. This movement is monitored easily by automated fluorescent microscopy and serves as the basis for screening libraries. Finally, secondary assays built into the cell system can differentiate between modulators that stimulate, inhibit, or do not affect the transcriptional activity of the receptor under study. This approach has been validated for both the estrogen receptor and the retinoic acid receptor and should be applicable to any member of the superfamily, facilitating the identification of new ligands and selective receptor modulators. PMID- 17110186 TI - The ligand-independent translocation assay: an enabling technology for screening orphan G protein-coupled receptors by arrestin recruitment. AB - Finding natural and/or synthetic ligands that activate orphan G protein-coupled receptors (oGPCRs) is a major focus in current drug discovery efforts. Transfluor is a cell-based GPCR screening platform that utilizes an arrestin-green fluorescent protein conjugate (arrestin-GFP) to detect ligand interactions with GPCRs. The assay is ideally suited for oGPCRs because binding of arrestin-GFP to activated receptors is independent of the interacting G protein. Before embarking on a high-throughput screen, it is important to know that the target oGPCR can actually bind arrestin-GFP. This information was thought to be inaccessible, however, as arrestin-GFP recruitment is an agonist-driven process. This chapter describes an assay that enables GPCRs to be validated in Transfluor in the absence of ligand. This assay, termed the ligand-independent translocation (LITe) assay, utilizes a modified G protein-coupled receptor kinase to bypass the requirement of ligand for initiating arrestin-GFP translocation. Using the LITe assay, one can determine if an oGPCR binds arrestin-GFP and if the response is quantifiable by high-content screening instruments. In addition, the assay expedites the development and identification of oGPCR stable cell lines with the best Transfluor properties. In this way, the assay provides criteria for selecting the best oGPCRs to move forward for a Transfluor screening campaign. Moreover, the assay can be used for quality control purposes during the orphan receptor screen itself by providing positive translocation responses for calculation of Z prime values. In summary, the LITe assay is a powerful new technology that enables a faster and more reliable path forward in the deorphanization of GPCRs with Transfluor. PMID- 17110187 TI - High-content screening of known G protein-coupled receptors by arrestin translocation. AB - G protein-coupled receptors (GPCRs) have proven to be one of the most successful target classes for drug discovery. Accordingly, many assays are available to screen GPCRs, including radioactive-binding assays, second messenger signaling assays, and downstream reporter assays. One of the more novel approaches is the Transfluor technology, a cell-based assay that uses a detectable tag on a cytosolic protein, called arrestin, that is involved in the desensitization or inactivation of GPCRs. Monitoring the translocation of GFP-tagged arrestin from the cytosol to activated GPCRs at the plasma membrane measures the pharmacological effect of test compounds that bind the receptor target. Moreover, the Transfluor assay provides further, high-content information on the test compound itself and its effects on cell processes due to the fluorescent imaging of whole cells used in this screen. Screening known GPCRs with Transfluor against large compound libraries is best accomplished in cell lines stably expressing an optimum level of the target receptor. This chapter describes how to generate a clonal cell line stably expressing the known GPCR with suitable Transfluor properties. It then describes the steps involved in performing a Transfluor screen and discusses high content data resulting from the screen. PMID- 17110188 TI - Cell imaging assays for G protein-coupled receptor internalization: application to high-throughput screening. AB - There are a number of assays currently available to study G protein-coupled receptors (GPCRs), including ligand binding and functional assays. The latter category, albeit more complex, offers some obvious advantages over traditional ligand-binding assays. Functional cell-based assays typically include second messenger and reporter gene assays, which depend directly or indirectly on the cellular signaling cascade initiated upon receptor activation, respectively. More recently, cell imaging assays monitoring receptor trafficking are becoming increasingly popular. These assays, described in greater detail in this chapter, are independent of receptor signaling and are thus ideally suited for orphan receptors. In addition, these assays provide a valuable measure of receptor desensitization, an important feature for the use of GPCR agonists as potential therapeutic agents. The most popular GPCR imaging assays are based on the principles of receptor desensitization and internalization monitored directly or indirectly by green fluorescent protein. PMID- 17110189 TI - High-throughput confocal microscopy for beta-arrestin-green fluorescent protein translocation G protein-coupled receptor assays using the Evotec Opera. AB - Ligand-activated G protein-coupled receptors (GPCRs) are known to regulate a myriad of homeostatic functions. Inappropriate signaling is associated with several pathophysiological states. GPCRs belong to a approximately 800 member superfamily of seven transmembrane-spanning receptor proteins that respond to a diversity of ligands. As such, they present themselves as potential points of therapeutic intervention. Furthermore, orphan GPCRs, which are GPCRs without a known cognate ligand, offer new opportunities as drug development targets. This chapter describes a systems-based biological approach, one that combines in silico bioinformatics, genomics, high-throughput screening, and high-content cell based confocal microscopy strategies to (1) identify a relevant subset of protein family targets, (2) within the therapeutic area of energy metabolism/obesity, (3) and to identify small molecule leads as tractable combinatorial and medicinal chemistry starting points. Our choice of screening platform was the Transfluor beta-arrestin-green fluorescent protein translocation assay in which full-length human orphan GPCRs were stably expressed in a U-2 OS cell background. These cells lend themselves to high-speed confocal imaging techniques using the Evotec Technologies Opera automated microscope system. The basic assay system can be implemented in any laboratory using a fluorescent probe, a stably expressed GPCR of interest, automation-assisted plate and liquid-handling techniques, an optimized image analysis algorithm, and a high-speed confocal microscope with sophisticated data analysis tools. PMID- 17110190 TI - G protein-coupled receptor internalization assays in the high-content screening format. AB - High-content screening (HCS), a combination of fluorescence microscopic imaging and automated image analysis, has become a frequently applied tool to study test compound effects in cellular disease-modeling systems. This chapter describes the measurement of G protein-coupled receptor (GPCR) internalization in the HCS format using a high-throughput, confocal cellular imaging device. GPCRs are the most successful group of therapeutic targets on the pharmaceutical market. Accordingly, the search for compounds that interfere with GPCR function in a specific and selective way is a major focus of the pharmaceutical industry today. This chapter describes methods for the ligand-induced internalization of GPCRs labeled previously with either a fluorophore-conjugated ligand or an antibody directed against an N-terminal tag of the GPCR. Both labeling techniques produce robust assay formats. Complementary to other functional GPCR drug discovery assays, internalization assays enable a pharmacological analysis of test compounds. We conclude that GPCR internalization assays represent a valuable medium/high-throughput screening format to determine the cellular activity of GPCR ligands. PMID- 17110191 TI - Screening for activators of the wingless type/Frizzled pathway by automated fluorescent microscopy. AB - Development of means to screen primary human cells rather than established cell lines is important in improving the predictive value of cellular assays in drug discovery. We describe a method of using automated fluorescent microscopy to detect activators of the wingless type/Frizzled (Wnt/Fzd) pathway in primary human preosteoblasts. This technique relies on detection of endogenous beta catenin translocation to the nucleus as an indicator of pathway activation, requires only a limited number of primary cells, and is robust enough for automation and high-content, high-throughput screening. Identification of activators of the Wnt/Fzd pathway in human preosteoblasts may be useful in providing lead compounds for the treatment of osteoporosis. PMID- 17110192 TI - A live cell, image-based approach to understanding the enzymology and pharmacology of 2-bromopalmitate and palmitoylation. AB - The addition of a lipid moiety to a protein increases its hydrophobicity and subsequently its attraction to lipophilic environments like membranes. Indeed most lipid-modified proteins are localized to membranes where they associate with multiprotein signaling complexes. Acylation and prenylation are the two common categories of lipidation. The enzymology and pharmacology of prenylation are well understood but relatively very little is known about palmitoylation, the most common form of acylation. One distinguishing characteristic of palmitoylation is that it is a dynamic modification. To understand more about how palmitoylation is regulated, we fused palmitoylation substrates to fluorescent proteins and reported their subcellular distribution and trafficking. We used automated high throughput fluorescence microscopy and a specialized computer algorithm to image and measure the fraction of palmitoylation reporter on the plasma membrane versus the cytoplasm. Using this system we determined the residence half-life of palmitate on the dipalmitoyl substrate peptide from GAP43 as well as the EC(50) for 2-bromopalmitate, a common inhibitor of palmitoylation. PMID- 17110193 TI - High-resolution, high-throughput microscopy analyses of nuclear receptor and coregulator function. AB - Steroid nuclear receptors are ligand-dependent transcription factors that have been studied since the early 1960s by principally biochemical and reporter assay approaches. From these studies an elegant and complex model of nuclear receptor transcription regulation has been developed. Inherent to both biochemical and reporter assay approaches is the generation of averaged responses and it is not generally considered that individual cells could exhibit quite varied responses. In some cases, recent microscopic single-cell analyses provide markedly different responses relative to traditional approaches based on population averaging and underscore the need to continue refinement of the current model of nuclear receptor-regulated transcription. While single-cell analyses of nuclear receptor action have been hindered by the predominantly qualitative nature of the approach, high-throughput microscopy is now available to resolve this issue. This chapter demonstrates the utility of high-throughput microscopic analyses of nuclear receptor and nuclear receptor coregulator function. The ability of high throughput microscopy to generate physiologically appropriate test populations by filtering based on morphological and protein of interest expression criteria is demonstrated. High-resolution, high-throughput microscopy is illustrated that provides quantitative subcellular information for both androgen and estrogen receptors. Efforts are ongoing to develop model systems that provide additional multiplex data and with refined image analyses to achieve true high-content imaging screens. PMID- 17110194 TI - Tracking individual proteins in living cells using single quantum dot imaging. AB - Single quantum dot imaging is a powerful approach to probe the complex dynamics of individual biomolecules in living systems. Due to their remarkable photophysical properties and relatively small size, quantum dots can be used as ultrasensitive detection probes. They make possible the study of biological processes, both in the membrane or in the cytoplasm, at a truly molecular scale and with high spatial and temporal resolutions. This chapter presents methods used for tracking single biomolecules coupled to quantum dots in living cells from labeling procedures to the analysis of the quantum dot motion. PMID- 17110195 TI - Development and application of automatic high-resolution light microscopy for cell-based screens. AB - Large-scale microscopy-based screens offer compelling advantages for assessing the effects of genetic and pharmacological modulations on a wide variety of cellular features. However, development of such assays is often confronted by an apparent conflict between the need for high throughput, which usually provides limited information on a large number of samples, and a high-content approach, providing detailed information on each sample. This chapter describes a novel high-resolution screening (HRS) platform that is able to acquire large sets of data at a high rate and light microscope resolution using specific "reporter cells," cultured in multiwell plates. To harvest extensive morphological and molecular information in these automated screens, we have constructed a general analysis pipeline that is capable of assigning scores to multiparameter-based comparisons between treated cells and controls. This chapter demonstrates the structure of this system and its application for several research projects, including screening of chemical compound libraries for their effect on cell adhesion, discovery of novel cytoskeletal genes, discovery of cell migration related genes, and a siRNA screen for perturbation of cell adhesion. PMID- 17110196 TI - Adenoviral sensors for high-content cellular analysis. AB - To maximize the potential of high-content cellular analysis for investigating complex cellular signaling pathways and processes, we have generated a library of adenoviral encoded cellular sensors based on protein translocation and reporter gene activation that enable a diverse set of assays to be applied to lead compound profiling in drug discovery and development. Adenoviral vector transduction is an efficient and technically simple system for expression of cellular sensors in diverse cell types, including primary cells. Adenoviral vector-mediated transient expression of cellular sensors, either as fluorescent protein fusions or live cell gene reporters, allows rapid assay development for profiling the activities of candidate drugs across multiple cellular systems selected for biological and physiological relevance to the target disease state. PMID- 17110197 TI - Cell-based assays using primary endothelial cells to study multiple steps in inflammation. AB - Cell-based assays are powerful tools for drug discovery and provide insight into complex signal transduction pathways in higher eukaryotic cells. Information gleaned from assays that monitor a cellular phenotype can be used to elucidate the details of a single pathway and to establish patterns of cross talk between pathways. By selecting the appropriate cell model, cell-based assays can be used to understand the function of a specific cell type in a complex disease process such as inflammation. We have used human umbilical vein endothelial cells to establish three cell-based, phenotypic assays that query different stages of a major signaling pathway activated in inflammation. One assay analyzes the tumor necrosis factor alpha (TNFalpha)-induced translocation of the transcription factor NF-kappaB from the cytoplasm into the nucleus 20 min after stimulation with TNFalpha. Two more assays monitor the expression of E-selectin and VCAM-1, 4 and 24 h after stimulation with TNFalpha. Indirect immunofluorescence and high throughput automated microscopy were used to analyze cells. Imaging was performed with the IN Cell Analyzer 3000. All assays proved to be highly robust. Z' values between 0.7 and 0.8 make each of the three assays well suited for use in high throughput screening for drug or probe discovery. PMID- 17110198 TI - Development and implementation of multiplexed cell-based imaging assays. AB - Fluorescence microscopy, image analysis, and automated screening technologies are some of the most powerful tools enabling cell biologists to investigate complex signaling pathways and compound or siRNA effects on cellular function in individual cells. Researchers can now use multiple fluorescent probes to quantify effects on intracellular molecular events, measure phenotypic changes, and provide contextual information about cellular pathways not discernible by traditional single-parameter, end point experiments. This chapter focuses on fluorescent labeling techniques and methods for designing image-based assays, multiplexed readouts, and image analysis routines. Case studies are presented describing the use of cell-based imaging assays for monitoring cell proliferation, cell cycle stage, and apoptosis. PMID- 17110199 TI - High-throughput screening for modulators of stem cell differentiation. AB - Realizing the potential of stem cell biology requires the modulation of self renewal and differentiation, both of which are incompletely understood. This chapter describes methods for the design, development, and implementation of cell based screens of small molecules, genes and expressed proteins for modulation of stem and progenitor cell fate. These include the engineering of embryonic and other stem cells with gene promoter-reporter protein constructs and their application in automated screening. We discuss considerations of promoter reporter selection, assay development and implementation, and image acquisition, analysis, and data handling. Such black-box screens are useful for the identification of probes of developmental processes and should provide tools that will identify druggable targets for biochemical assays. PMID- 17110200 TI - High-content kinetic calcium imaging in drug-sensitive and drug-resistant human breast cancer cells. AB - Intracellular calcium (Ca2+) is involved in the regulation of a variety of biological functions in cancer cells, including growth inhibition, tumor invasiveness, and drug resistance. To gain insight into the possible role played by Ca2+ in the development of drug resistance in breast cancer, we performed a comparative high-content analysis of the intracellular Ca2+ dynamics in drug sensitive human breast cancer MCF-7 cells and five drug-resistant, MCF-7-derived clonal cell lines. Fura-2 single cell ratiometric fluorescence microscopy was used to monitor real-time quantitative changes in cytosolic-free Ca2+ concentration ( [Ca2+]i ) upon addition of phosphoinositol-coupled receptor agonists. While the magnitude and the onset kinetics of the [Ca2+]i rise were similar in drug-sensitive and drug-resistant cell lines, the decay kinetics of the [Ca2+]i increase was found to be consistently slower in drug-resistant than drug-sensitive cells. Such a delay in reestablishing homeostatic [Ca2+]i persisted in the absence of extracellular Ca2+ and was independent of the expression or function of specific drug efflux pumps associated with drug resistance. Moreover, intracellular Ca2+ pools releasable by phosphoinositol coupled receptor agonists or thapsigargin appeared to be differentially shared in drug-sensitive and drug-resistant cells. In light of the clinical relevance that drug resistance has in the treatment of cancer, the molecular and biochemical relationship between alterations in Ca2+ dynamics and drug resistance demands to be further investigated and tested in a wider array of cell types. Automated microscopy will help greatly in this pursuit by facilitating both sample imaging and data analysis, thus allowing high-content as well as high-throughput screening of large sample sets. A protocol for studying [Ca2+]i kinetics with a commercially available automated imaging platform is described. PMID- 17110201 TI - Measurement and analysis of calcium signaling in heterogeneous cell cultures. AB - High-content imaging platforms capable of studying kinetic responses at a single cell level have elevated kinetic recording techniques from labor-intensive low throughput experiments to potential high-throughput screening assays. We have applied this technology to the investigation of heterogeneous cell cultures derived from primary neural tissue. The neuronal cultures mature into a coupled network and display spontaneous oscillations in intracellular calcium, which can be modified by the addition of pharmacological agents. We have developed algorithms to perform Fourier analysis and quantify both the degree of synchronization and the effects of modulators on the oscillations. Functional and phenotypic experiments can be combined using this approach. We have used post-hoc immunolabeling to identify subpopulations of cells in cocultures and to dissect the calcium responses of these cells from the population response. The combination of these techniques represents a powerful tool for drug discovery. PMID- 17110202 TI - Multiplex analysis of inflammatory signaling pathways using a high-content imaging system. AB - This chapter describes a robust high-content cellular screening assay to simultaneously analyze the spatiotemporal activation of three different kinase associated signaling pathways involving NF-kappaB, JNK, and p38, all of which are closely implicated in proliferative and proinflammatory responses. Signal transduction is dependent on the translocation of NF-kappaB p65 and phosphorylated c-Jun and p38 from the cytosol to the nucleus, and fluorescent immunolabeling was used to monitor changes in their cellular distribution. Cellular screening, data acquisition, and data interpretation were conducted on the ArrayScan HCS Reader (Cellomics Inc., Pittsburgh, PA). Assay adaptation to various cellular systems is feasible when sufficient separation of the nuclear and cytosolic compartment can be achieved and if cell adhesion properties permit proper attachment to the culture plates. Substitution of NF-kappaB p65 and phosphorylated forms of c-Jun and p38 as targets to analyze other translocating components is possible and is limited primarily by antibody specificity and the risk of fluorescent bleed-through between emission channels. Because assay validity is particularly confounded by inadequate spectral separation of the detection dyes in multicolor labeling assays, means of eliminating or counterbalancing staining artifacts are illustrated. Also, protocol parameter settings important for imaging and image processing are described, including object identification, image exposure settings, separation of cytosolic and nuclear regions, number of cells sufficient for analysis, and the use of gating thresholds critical for cell sorting and subpopulation analysis. This assay is a useful tool to investigate the interplay between signaling pathways and the mode of action, potency, and selectivity of compound inhibition of specific target molecules in a cellular context. PMID- 17110203 TI - Generation and characterization of a stable MK2-EGFP cell line and subsequent development of a high-content imaging assay on the Cellomics ArrayScan platform to screen for p38 mitogen-activated protein kinase inhibitors. AB - This chapter describes the generation and characterization of a stable MK2-EGFP expressing HeLa cell line and the subsequent development of a high-content imaging assay on the Cellomics ArrayScan platform to screen for p38 MAPK inhibitors. Mitogen-activated protein kinase activating protein kinase-2 (MK2) is a substrate of p38 MAPK kinase, and p38-induced phosphorylation of MK-2 induces a nucleus to cytoplasm translocation (Engel et al., 1998; Neininger et al., 2001; Zu et al., 1995). Through a process of heterologous expression of a MK2-EGFP fusion protein in HeLa cells using retroviral infection, antibiotic selection, and flow sorting, we were able to isolate a cell line in which the MK2-EGFP translocation response could be robustly quantified on the Cellomics ArrayScan platform using the nuclear translocation algorithm. A series of assay development experiments using the A4-MK2-EGFP-HeLa cell line are described to optimize the assay with respect to cell seeding density, length of anisomycin stimulation, dimethyl sulfoxide tolerance, assay signal window, and reproducibility. The resulting MK2-EGFP translocation assay is compatible with high-throughput screening and was shown to be capable of identifying p38 inhibitors. The MK2-EGF translocation response is susceptible to other classes of inhibitors, including nonselective kinase inhibitors, kinase inhibitors that inhibit upstream kinases in the p38 MAPK signaling pathway, and kinases involved in cross talk between different modules (ERKs, JNKs, and p38s) of the MAPK signaling pathways. An example of mining "high-content" image-based multiparameter data to extract additional information on the effects of compound treatment of cells is presented. PMID- 17110204 TI - Development and implementation of three mitogen-activated protein kinase (MAPK) signaling pathway imaging assays to provide MAPK module selectivity profiling for kinase inhibitors: MK2-EGFP translocation, c-Jun, and ERK activation. AB - This chapter describes the development and implementation of three independent imaging assays for the major mitogen-activated protein kinase (MAPK) signaling modules: p38, JNK, and ERK. There are more than 500 protein kinases encoded in the human genome that share an ATP-binding site and catalytic domain conserved in both sequence and structure. The majority of kinase inhibitors have been found to be competitive with ATP, raising concerns regarding kinase selectivity and potency in an environment of millimolar intracellular concentrations of ATP, as well as the potential for off-target effects via the many other cellular proteins that bind and/or utilize ATP. The apparent redundancy of the kinase isoforms and functions in the MAPK signaling modules present additional challenges for kinase inhibitor selectivity and potency. Imaging assays provide a method to address many of these concerns. Cellular imaging approaches facilitate analysis of the targets expressed in the context of their endogenous substrates and scaffolding proteins and in a complex environment for which subcellular localization, cross talk between pathways, phosphatase regulatory control, and intracellular ATP concentrations are relevant to the functions of the kinase. The assays described herein provide a strategy to profile kinase inhibitors for MAPK pathway selectivity while simultaneously providing information on cell morphology or toxicity. Results suggest that the MAPK pathways are indeed susceptible to nonselective kinase inhibitors such as staurosporin and inhibitors that inhibit upstream MAPK Kinase Kinases (MKKKs) and MAPK Kinases (MKKs) in the MAPK signaling pathway, especially those involved in cross talk between the pathways. However, selective MAPK inhibitors were identified that exhibited pathway selectivity as evidenced by significantly lower IC(50) values for their respective p38, JNK, or ERK signaling pathway assays. PMID- 17110205 TI - Assay development and case history of a 32K-biased library high-content MK2-EGFP translocation screen to identify p38 mitogen-activated protein kinase inhibitors on the ArrayScan 3.1 imaging platform. AB - This chapter describes the conversion and assay development of a 96-well MK2-EGFP translocation assay into a higher density 384-well format high-content assay to be screened on the ArrayScan 3.1 imaging platform. The assay takes advantage of the well-substantiated hypothesis that mitogen-activated protein kinase activating protein kinase-2 (MK2) is a substrate of p38 MAPK kinase and that p38 induced phosphorylation of MK-2 induces a nucleus-to-cytoplasm translocation. This chapter also presents a case history of the performance of the MK2-EGFP translocation assay, run as a "high-content" screen of a 32K kinase-biased library to identify p38 inhibitors. The assay performed very well and a number of putative p38 inhibitor hits were identified. Through the use of multiparameter data provided by the nuclear translocation algorithm and by checking images, a number of compounds were identified that were potential artifacts due to interference with the imaging format. These included fluorescent compounds, or compounds that dramatically reduced cell numbers due to cytotoxicity or by disrupting cell adherence. A total of 145 compounds produced IC(50) values <50.0 muM in the MK2-EGFP translocation assay, and a cross target query of the Lilly RTP HTS database confirmed their inhibitory activity against in vitro kinase targets, including p38a. Compounds were confirmed structurally by LCMS analysis and profiled in cell-based imaging assays for MAPK signaling pathway selectivity. Three of the hit scaffolds identified in the MK2-EGFP translocation HCS run on the ArrayScan were selected for a p38a inhibitor hit-to-lead structure activity relationship (SAR) chemistry effort. PMID- 17110206 TI - Compound classification using image-based cellular phenotypes. AB - Compounds with similar target specificities and modes of inhibition cause similar cellular phenotypes. Based on this observation, we hypothesized that we could quantitatively classify compounds with diverse mechanisms of action using cellular phenotypes and identify compounds with unintended cellular activities within a chemical series. We have developed Cytometrix technologies, a highly automated image-based system capable of quantifying, clustering, and classifying changes in cellular phenotypes for this purpose. Using this system, 45 out of 51 known compounds were accurately classified into 12 distinct mechanisms of action. We also demonstrate microtubule-binding activity in one of seven related cytochalasin actin poisons. This technology can be used for a variety of drug discovery applications, including high-throughput primary screening of chemical and siRNA libraries and as a secondary assay to detect unintended activities and toxicities. PMID- 17110207 TI - High-content screening: emerging hardware and software technologies. AB - The field of high-content screening has flourished since 2000 with advancements in automated fluorescence microscopy technologies, fluorescent labeling techniques, and sophisticated image analysis software. Through the use of these technologies, researchers can now monitor cellular and molecular events in individual cells in vitro following drug treatment or RNAi and rapidly screen compound and siRNA libraries. This chapter discusses current and next-generation hardware and software features and capabilities. PMID- 17110208 TI - An infrastructure for high-throughput microscopy: instrumentation, informatics, and integration. AB - High-throughput, image-based cell assays are rapidly emerging as valuable tools for the pharmaceutical industry and academic laboratories for use in both drug discovery and basic cell biology research. Access to commercially available assay reagents and automated microscope systems has made it relatively straightforward for a laboratory to begin running assays and collecting image-based cell assay data, but doing so on a large scale can be more challenging. Challenges include process bottlenecks with sample preparation, image acquisition, and data analysis as well as day-to-day assay consistency, managing unprecedented quantities of image data, and fully extracting useful information from the primary assay data. This chapter considers many of the decisions needed to build a robust infrastructure that addresses these challenges. Infrastructure components described include integrated laboratory automation systems for sample preparation and imaging, as well as an informatics infrastructure for multilevel image and data analysis. Throughout the chapter we describe a variety of strategies that emphasize building processes that are scaleable, highly efficient, and rigorously quality controlled. PMID- 17110209 TI - Protein translocation assays: key tools for accessing new biological information with high-throughput microscopy. AB - Redistribution technology is a cell-based assay technology that uses protein translocation as the primary readout for the activity of cellular signaling pathways and other intracellular events. Protein targets are labeled with the green fluorescent protein, and stably transfected cell lines are generated. The assays are read using a high-throughput, optical microscope-based instrument, several of which have become available commercially. Protein translocation assays can be formatted as agonist assays, in which compounds are tested for their ability to promote protein translocation, or as antagonist assays, in which compounds are tested for their ability to inhibit protein translocation caused by a known agonist. Protein translocation assays are high-content, high-throughput assays primarily used for profiling of lead series, primary screening of compound libraries, and as readouts for gene-silencing studies using siRNAs. This chapter describes two novel high-content Redistribution assay technologies: (1) The p53:hdm2 GRIP interaction assay, in which one high-content image feature is used for detection of primary hits, whereas a different feature is used to deselect compounds with unwanted mode of action, and (2) application of siRNAs to Redistribution assays, exemplified by knockdown of Akt isoforms in a FKHR translocation assay reporting on the PI3 kinase signaling pathway. PMID- 17110210 TI - High-content screening of functional genomic libraries. AB - Recent advances in functional genomics have enabled genome-wide genetic studies in mammalian cells. These include the establishment of high-throughput transfection and viral propagation methodologies, the production of large-scale cDNA and siRNA libraries, and the development of sensitive assay detection processes and instrumentation. The latter has been significantly facilitated by the implementation of automated microscopy and quantitative image analysis, collectively referred to as high-content screening (HCS), toward cell-based functional genomics application. This technology can be applied to whole genome analysis of discrete molecular and phenotypic events at the level of individual cells and promises to significantly expand the scope of functional genomic analyses in mammalian cells. This chapter provides a comprehensive guide for curating and preparing function genomics libraries and performing HCS at the level of the genome. PMID- 17110211 TI - Fluorescent protein-based cellular assays analyzed by laser-scanning microplate cytometry in 1536-well plate format. AB - Microtiter plate readers have evolved from photomultiplier and charged-coupled device-based readers, where a population-averaged signal is detected from each well, to microscope-based imaging systems, where cellular characteristics from individual cells are measured. For these systems, speed and ease of data analysis are inversely proportional to the amount of data collected from each well. Microplate laser cytometry is a technology compatible with a 1536-well plate format and capable of population distribution analysis. Microplate cytometers such as the Acumen Explorer can monitor up to four fluorescent signals from single objects in microtiter plates with densities as high as 1536 wells. These instruments can measure changes in fluorescent protein expression, cell shape, or simple cellular redistribution events such as cytoplasmic to nuclear translocation. To develop high-throughput screening applications using laser scanning microplate cytometry, we used green fluorescent protein- and yellow fluorescent protein-expressing cell lines designed to measure diverse biological functions such as nuclear translocation, epigenetic signaling, and G protein coupled receptor activation. This chapter illustrates the application of microplate laser cytometry to these assays in a manner that is suitable for screening large compound collections in high throughput. PMID- 17110212 TI - High-throughput measurements of biochemical responses using the plate::vision multimode 96 minilens array reader. AB - The plate::vision is a high-throughput multimode reader capable of reading absorbance, fluorescence, fluorescence polarization, time-resolved fluorescence, and luminescence. Its performance has been shown to be quite comparable with other readers. When the reader is integrated into the plate::explorer, an ultrahigh-throughput screening system with event-driven software and parallel plate-handling devices, it becomes possible to run complicated assays with kinetic readouts in high-density microtiter plate formats for high-throughput screening. For the past 5 years, we have used the plate::vision and the plate::explorer to run screens and have generated more than 30 million data points. Their throughput, performance, and robustness have speeded up our drug discovery process greatly. PMID- 17110213 TI - Systems cell biology based on high-content screening. AB - A new discipline of biology has emerged since 2004, which we call "systems cell biology" (SCB). Systems cell biology is the study of the living cell, the basic unit of life, an integrated and interacting network of genes, proteins, and myriad metabolic reactions that give rise to function. SCB takes advantage of high-content screening platforms, but delivers more detailed profiles of cellular systemic function, including the application of advanced reagents and informatics tools to sophisticated cellular models. Therefore, an SCB profile is a cellular systemic response as measured by a panel of reagents that quantify a specific set of biomarkers. PMID- 17110214 TI - Digital autofocus methods for automated microscopy. AB - Automatic focusing of microscope images is an essential part of modern high throughput microscopy. This chapter describes implementation of a robust autofocus system appropriate for using either air or oil immersion objectives in robotic imaging. Both hardware and software algorithms are described, and caveats of using viscous immersion media with multifield scanning are detailed. PMID- 17110215 TI - Fluorescence lifetime imaging microscopy: two-dimensional distribution measurement of fluorescence lifetime. AB - A newly developed fluorescence lifetime imaging microscopy (FLIM) system has combined the high-temporal resolution of a streak camera with the high-spatial resolution of a microscope to obtain a two-dimensional distribution of fluorescence lifetimes within living cells. The temporal resolution is as short as 20 ps. The effective field of view is 48 x 45 mum with 0.2 mum resolution using a 60x water immersion objective. Image acquisition time is as short as 3 s per image. Measured and published values of lifetime for standard fluophores are shown with good agreement. Examples of FLIM and fluorescence resonance energy transfer images are presented. PMID- 17110216 TI - Coccidioidomycosis: a review and update. AB - Coccidioidomycosis occurs in arid and semi-arid regions of the New World from the western United States to Argentina. Highly endemic areas are present in the southwest United States. Coccidioides species live in the soil and produce pulmonary infection via airborne arthroconidia. The skin may be involved by dissemination of the infection, or by reactive eruptions, such as a generalized exanthem or erythema nodosum. Interstitial granulomatous dermatitis and Sweet's syndrome have recently been recognized as additional reactive signs of the infection. Coccidioidomycosis is a "great imitator" with protean manifestations. Cutaneous findings may be helpful clues in the diagnosis of this increasingly important disease. PMID- 17110217 TI - The importance of dual 5alpha-reductase inhibition in the treatment of male pattern hair loss: results of a randomized placebo-controlled study of dutasteride versus finasteride. AB - BACKGROUND: Male pattern hair loss (MPHL) is a potentially reversible condition in which dihydrotestosterone is an important etiologic factor. OBJECTIVE: Our aim was to evaluate the efficacy of the type 1 and 2 5alpha-reductase inhibitor dutasteride in men with MPHL. METHODS: Four hundred sixteen men, 21 to 45 years old, were randomized to receive dutasteride 0.05, 0.1, 0.5 or 2.5 mg, finasteride 5 mg, or placebo daily for 24 weeks. RESULTS: Dutasteride increased target area hair count versus placebo in a dose-dependent fashion and dutasteride 2.5 mg was superior to finasteride at 12 and 24 weeks. Expert panel photographic review and investigator assessment of hair growth confirmed these results. Scalp and serum dihydrotestosterone levels decreased, and testosterone levels increased, in a dose-dependent fashion with dutasteride. LIMITATIONS: The study was limited to 24 weeks. CONCLUSION: Dutasteride increases scalp hair growth in men with MPHL. Type 1 and type 2 5alpha-reductase may be important in the pathogenesis and treatment of MPHL. PMID- 17110218 TI - Advancement flap for the reconstruction of nasal ala and lateral nasal tip defects. AB - Repair of nasal ala and lateral nasal tip defects provide unique reconstructive challenges. This article describes a one-staged advancement flap for repair of such defects. The flap may be medially-based, laterally-based, or bilaterally based. Sharp undermining is recommended, and a standing cone must be removed superior to the defect, perpendicular to the alar rim. Temporary alar rim flattening is accepted, and normalizes with time. This reconstruction has provided excellent functional and cosmetic results for defects measuring <1 cm in diameter on the nasal ala and lateral nasal tip. PMID- 17110219 TI - Self-healing juvenile cutaneous mucinosis: cases highlighting subcutaneous/fascial involvement. AB - BACKGROUND: Self-healing juvenile cutaneous mucinosis is a rare disease affecting young people characterized by transient cutaneous lesions and sometimes mild inflammatory symptoms. The deep dermal and subcutaneous features of this disorder have not yet been well described. OBJECTIVE: The purpose of our study was to present 3 cases of self-healing juvenile cutaneous mucinosis in which the histopathologic features caused diagnostic confusion between this disorder and proliferative fasciitis. METHODS: The study includes clinical and histologic findings of 3 patients, complemented by a literature review. RESULTS: The histologic descriptions of nodular lesions in self-healing juvenile cutaneous mucinosis reveal features of proliferative fasciitis, including a myxoid stroma and gangliocyte-like giant cells. LIMITATIONS: Self-healing juvenile cutaneous mucinosis is a rare condition and has not been frequently reported in medical literature. Our findings are based on the pathologic features of 3 patients. CONCLUSIONS: Our findings further elucidate the histologic features of self healing juvenile cutaneous mucinosis and expand the differential diagnosis for entities in which gangliocyte-like giant cells are noted. PMID- 17110220 TI - Clinically occult cutaneous metastases. AB - Cutaneous metastases present themselves in a variety of clinical patterns and tend to be manifested as indurated papules/nodules/tumors. Some of those clinical expressions are unique for certain types of metastases. This report describes an entirely different phenomenon of clinically incognito cutaneous metastases that were only apparent histopathologically as an incidental finding. PMID- 17110221 TI - PHACES association. PMID- 17110222 TI - Flushing, pheochromocytoma, and the dermatologist. PMID- 17110223 TI - Medical pearl: intraoral medication application using the "parked" cotton ball technique. PMID- 17110224 TI - Medical pearl: circle of desquamation--a clue to the diagnosis of folliculitis and furunculosis caused by Staphylococcus aureus. PMID- 17110225 TI - Hematoma-like metastases. PMID- 17110226 TI - Pemphigus foliaceus treated with etanercept. PMID- 17110227 TI - Chylous reflux presenting with minimal milia-like lesions on the thigh. PMID- 17110228 TI - How to deal with a tomato which has undergone surgery. PMID- 17110229 TI - Tinea capitis in the United States: Diagnosis, treatment, and costs. PMID- 17110230 TI - Methods for diagnosing dermatitis herpetiformis. PMID- 17110232 TI - Estrogen and the skin. PMID- 17110233 TI - Multidrug therapy regimen for leprosy. PMID- 17110234 TI - Genital human papillomavirus-associated (pre-) malignant skin diseases drastically increase in the era of highly active antiretroviral therapy for HIV infection. PMID- 17110236 TI - Standard lipid profile. AB - The standard lipid profile has an enormous scientific evidence base and has provided simple clinical lipid-altering goals focused on low-density lipoprotein (LDL) cholesterol that have been shown to reduce coronary heart disease. Its limitations in estimating LDL cholesterol are well known: increasing inaccuracies in the nonfasting state, in individuals with triglycerides greater than 200 to 400 mg/dL, and as an accurate measure of LDL particle risk in size and concentration. Until studies are done comparing the benefit of targeting LDL particle concentration and size versus ever lower LDL cholesterol concentration, widespread clinical use of continuing revisions of the standard lipid profile goals will continue to remain one of the giant clinical and public health achievements of the last 50 years. PMID- 17110237 TI - Apolipoprotein A1 and B. AB - This article reviews the evidence showing that apolipoprotein (apo) B and A-1 are superior to the conventional cholesterol indices as analytes in laboratory practice, indices of the lipoprotein-related risk for vascular disease, and measures of the adequacy of low-density lipoprotein-lowering therapy. PMID- 17110238 TI - Lipoprotein(a): a unique risk factor for cardiovascular disease. AB - Lipoprotein(a) (Lp(a)) is present in humans and primates. It has many properties in common with low-density lipoprotein, but contains a unique protein moiety designated apo(a), which is linked to apolipoprotein B-100 by a single disulfide bond. International standards for Lp(a) measurement and optimized Lp(a) assays insensitive to isoform size are not yet widely available. Lp(a) is a risk factor for coronary artery disease, and smaller size apo(a) is associated with coronary artery disease. The physiologic role of Lp(a) is unknown. PMID- 17110239 TI - Postprandial lipemia and remnant lipoproteins. AB - Increased postprandial lipemia or elevated levels of triglyceride-rich remnant lipoproteins in fasting plasma are associated with increased risk of coronary artery disease. Despite many studies showing that postprandial triglyceride-rich lipoproteins play a central role in the pathogenesis of atherosclerosis, suitably standardized methods to measure postprandial lipemia or remnant lipoproteins in the clinical setting are lacking. This approach for cardiovascular risk assessment is confined to research laboratories and for the time being is not a standard procedure in clinical practice. PMID- 17110240 TI - Cholesterol profile measurement by vertical auto profile method. AB - Vertical auto profile (VAP) method is a direct single test for measuring comprehensive lipoprotein cholesterol profile. It is based on a well-established method of ultracentrifugation that uses vertical rotor and single density gradient spin. VAP provides cholesterol concentrations of total lipoprotein, high density lipoprotein (HDL), low-density lipoprotein (LDL), very-low-density lipoprotein (VLDL), lipoprotein(a) (Lp(a)), intermediate-density lipoprotein (IDL), HDL subclasses (HDL2 and HDL3), LDL subclasses (LDL1, LDL2, LDL3, and LDL4), VLDL subclasses (VLDL1, VLDL2, and VLDL3), and LDL maximum time, which is directly proportional to LDL size. Because VAP measures additional lipoprotein classes, such as Lp(a), IDL, and subclasses of HDL, LDL, and VLDL, it can identify patients at high risk for coronary heart disease who cannot be identified using the standard lipid panel. In addition, the VAP method is compliant with the National Cholesterol Education Program's Adult Treatment Panel III guidelines. PMID- 17110241 TI - Polyacrylamide gradient gel electrophoresis of lipoprotein subclasses. AB - High-density (HDL), low-density (LDL), and very-low-density (VLDL) lipoproteins are heterogeneous cholesterol-containing particles that differ in their metabolism, environmental interactions, and association with disease. Several protocols use polyacrylamide gradient gel electrophoresis (GGE) to separate these major lipoproteins into known subclasses. This article provides a brief history of the discovery of lipoprotein heterogeneity and an overview of relevant lipoprotein metabolism, highlighting the importance of the subclasses in the context of their metabolic origins, fates, and clinical implications. Various techniques using polyacrylamide GGE to assess HDL and LDL heterogeneity are described, and how the genetic and environmental determinations of HDL and LDL affect lipoprotein size heterogeneity and the implications for cardiovascular disease are outlined. PMID- 17110242 TI - Lipoprotein particle analysis by nuclear magnetic resonance spectroscopy. AB - Laboratory measurements of plasma lipids (principally cholesterol and triglycerides) and lipoprotein lipids (principally low-density lipoprotein [LDL] and low-density lipoprotein [HDL] cholesterol) are the cornerstone of the clinical assessment and management of atherosclerotic cardiovascular disease (CVD) risk. LDL particles, and to a lesser extent very-low-density lipoprotein [VLDL] particles, cause atherosclerosis, whereas HDL particles prevent or reverse this process through reverse cholesterol transport. The overall risk for CVD depends on the balance between the "bad" LDL (and VLDL) and "good" HDL particles. Direct assessment of lipoprotein particle numbers us now possible through nuclear magnetic resonance spectroscopic analysis. PMID- 17110245 TI - Are you attacking the correct entity? PMID- 17110247 TI - Consider the evidence. PMID- 17110248 TI - Skeletal and soft-tissue response to genioplasty. PMID- 17110251 TI - Comparison of frictional resistance after immersion of metal brackets and orthodontic wires in a fluoride-containing prophylactic agent. AB - INTRODUCTION: In this study, we investigated and compared the levels of frictional resistance between metal brackets and orthodontic wires after immersion in an acidified phosphate fluoride (APF) agent. METHODS: Three types of mandibular incisor stainless steel metal brackets with beta-titanium alloy wire (TMA), heat-activated nickel-titanium wire (Ni-Ti), and 2 sizes of stainless steel wires (SSW) were immersed in 0.2% APF and pH 6.75 artificial saliva solutions for 24 hours. The study included 480 bracket-wire specimens. The frictional resistance was measured on an EZ-test machine (Shimadazu, Tokyo, Japan) with a 5-N load cell. An Alastik (Quik-Stik Clear, A-1 Alastik, 3M Unitek, Monrovia, Calif) module ligated to the bracket was attached to the crosshead of the machine and pulled at a speed of 10 mm per minute for a distance of 5 mm. A completely randomized 1-way ANOVA was used to test for significant differences among the 3 bracket/wire specimens after immersion in 0.2% APF and pH 6.75 artificial saliva solutions. This was followed by the Student-Newman-Keuls multiple comparison of means ranking at P <.05 to determine differences between the groups. RESULTS: In the APF-immersed group, the static frictional force was greater than the kinetic frictional force. The frictional forces of the orthodontic wires had statistically significant differences (P <.05) in this progressive order: TMA, Ni-Ti, and SSW. Similar frictional force results were obtained in the pH 6.75 saliva group (P <.05). The frictional force values of the APF group were higher than those of the pH 6.75 saliva group (P <.05). CONCLUSIONS: This study demonstrates that the frictional forces of orthodontic brackets and wires are influenced by contact with fluoride-containing solutions. PMID- 17110252 TI - Levels of matrix metalloproteinases 1 and 2 in human gingival crevicular fluid during initial tooth movement. AB - INTRODUCTION: During orthodontic treatment, the early response of periodontal tissues to mechanical stress involves several metabolic changes that allow tooth movement. Many studies have evaluated these modifications through the analysis of various metabolites released into gingival crevicular fluid (GCF). The purpose of this investigation was to evaluate matrix metalloproteinase (MMP)-1 and MMP-2 in the GCF of human teeth exposed to orthodontic force on both the tension and compression sides in the initial phase of orthodontic tooth movement. METHODS: GCF samples were obtained from 11 healthy orthodontic patients (8 girls, 3 boys; age, 13-15 years; mean, 13.9 years) who needed their 4 first premolars extracted for orthodontic reasons. In each patient, the left maxillary canine having the fixed orthodontic appliance was used as the test tooth, and its antagonist, with no appliance, was the control tooth. Orthodontic force was applied by using a Sentalloy coil-spring (GAC International, Bohemia, NY) of 150 g. The GCF sampling on the mesiobuccal and distobuccal aspects of each experimental and control tooth was performed at specific times up to 8 hours with paper strips. Processing was carried out with western blot analysis to detect MMP-1 and MMP-2 levels on the compression and tension sides. RESULTS: Compression force induced a significant increase of MMP-1 protein after 1 hour; the increase lasted until the third hour of force application and disappeared thereafter. The tension force induced significantly increased levels of the MMP-1 protein after just 1 hour of force application. MMP-2 protein was induced by compression and increased significantly in a time-dependent fashion, reaching a peak after 8 hours of force application. On the tension side, MMP-2 was significantly increased after 1 hour but gradually returned to basal levels within 8 hours. CONCLUSIONS: Orthodontic forces affect both MMP-1 and MMP-2 protein levels on the compression and the tension sides, although to different extents, whereas MMP-1 and MMP-2 protein levels change in a time-dependent fashion. PMID- 17110253 TI - Body dysmorphic disorder in adult orthodontic patients. AB - INTRODUCTION: Body image plays an important role for patients seeking orthodontic treatment. It affects how patients feel about their physical appearance and, in extreme cases, can lead to subjective fears of ugliness. When there is a physical defect that, although within normal limits, seems far more noticeable to the patient, this may be diagnosed as body dysmorphic disorder (BDD). This interview based study was designed to assess BDD in adults attending the Orthodontic Department at the Eastman Dental Hospital in London for their initial consultations for orthodontic treatment and also in the general public. METHODS: A total of 70 members of the general public and 40 patients, all over 18 years of age, were assessed. The BDD modification of the Yale-Brown obsessive compulsive scale was used for diagnosis of BDD. RESULTS: BDD was diagnosed in 2 members of the general public (2.86%) and 3 patients (7.5%). CONCLUSIONS: It is important to have an understanding of body image and to be able to identify orthodontic patients who have BDD. These patients are rarely satisfied with the results of treatment, and it is therefore important to recognize them to avoid unnecessary treatment and to refer them for appropriate management. PMID- 17110254 TI - Scanning electron microscopic evaluation of enamel surfaces exposed to 3 orthodontic bonding systems. AB - INTRODUCTION: This in-vitro study investigated the effects of conventional etching with a 35% phosphoric etching gel and priming/bonding with Transbond XT primer/adhesive (3M Unitek, Monrovia, Calif), conditioning with 10% polyacrylic acid and bonding with a resin-modified glass ionomer cement (Fuji ORTHO LC, GC Corporation, Tokyo Japan) or using a self-etching bonding system (Transbond Plus) and bonding with Transbond XT adhesive on the surface morphology of the enamel. The study was conducted at the Faculty of Dentistry, University of Oslo, in Norway, 2004. METHODS: The enamel surfaces of extracted premolars where conditioned with 1 of the 3 systems and evaluated with environmental scanning electron microscope. Brackets were then bonded with 1 of 3 bonding agents and examined with a conventional scanning electron microscope. RESULTS: The bonding systems induced different effects on the enamel structure. Phosphoric acid etching produced a rough, etched surface with the typical honeycomb pattern. Bonding brackets to such a surface resulted in thick resin tags relatively deep into the enamel. Less pronounced etching of the surface enamel was obtained by the self-etching primer system, and bonding resulted in smaller and fewer resin tags. The resin-modified glass ionomer bonded after conditioning the enamel with polyacrylic acid showed no resin tags. CONCLUSIONS: Bonding systems with self etching primers or conditioners with polyacrylic acid might offer potential benefits compared with conventional acid etching and priming because of fewer irreversible changes to the enamel surface. PMID- 17110255 TI - Treatment effects of the edgewise Herbst appliance: a cephalometric and tomographic investigation. AB - INTRODUCTION: The crown Herbst appliance was introduced in the late 1980s because of shortcomings of the banded Herbst. In edgewise Herbst treatment, a fixed appliance is used with the crown Herbst to maximize the skeletal effects of treatment. Treatment response to the edgewise Herbst appliance has not been reported in the literature. Our objective was to investigate skeletal and dental changes in patients with Class II malocclusions treated with the edgewise Herbst appliance. METHODS: Fifty-two consecutive patients were treated with the edgewise Herbst appliance; 32 (18 girls, 14 boys) met the criterion of 16 months out of Herbst treatment and were included in the study. Mean treatment time with this appliance was 8.0 +/- 1.8 months. Patients in the mixed dentition received additional treatment with 2 x 4 appliances until proper overbite, overjet, and torque on the incisors and permanent first molars were achieved. Patients in the permanent dentition were treated with full appliances to finalize the occlusion. Cephalometric measurements were taken at pretreatment, posttreatment, and 16 months after removal of the Herbst appliance, and the results were compared with 32 untreated Class II subjects from the Bolton Brush Study, matched for sex, age, and cephalometric dentofacial morphology. Data were analyzed with ANOVA, Tukey Kramer multiple comparison tests, and 2-tailed t tests. RESULTS: After 8 months of Herbst treatment, incisal relationship was overcorrected to an end-to-end incisal relationship and improved 8.4 mm, compared with the control group. The maxilla moved backward 1.4 mm at Point A, and the mandible moved forward 1.7 mm. The maxillary incisors moved lingually 1.7 mm, and the mandibular incisors were proclined 3.6 mm. The molars were corrected to a Class III relationship with a change of 7.2 mm compared with the control group. The mandible moved downward and forward. However, the condyle showed only 0.2 mm forward movement in the fossa. Sixteen months after appliance removal, the molars had relapsed into a Class I relationship, for a net change of 2.4 mm compared with the control group. Net overjet gain was 2.7 mm. Net restraint of maxillary growth was 1.3 mm, and net forward movement of the mandible was 1.0 mm. The maxillary incisors had no net movement, and the mandibular incisors had a net forward movement of 0.3 mm. Overall, skeletal change contributed 85% of the net overjet correction. CONCLUSIONS: Class II treatment with the edgewise Herbst appliance is accompanied by both skeletal and dental changes. The changes are stable, with significant skeletal differences remaining 16 months after appliance removal. The forward and downward movement of the mandible with minimal changes in the position of the condyles in the fossae suggests a combination of condylar growth and remodeling of the glenoid fossa with treatment. PMID- 17110256 TI - Skeletal and dentoalveolar effects of Twin-block and bionator appliances in the treatment of Class II malocclusion: a comparative study. AB - INTRODUCTION: The purpose of this study was to evaluate the skeletal and dentoalveolar effects of the Twin-block and bionator appliances in the treatment of Class II Division 1 malocclusions. METHODS: Fifty-five girls from North India with Class II Division 1 malocclusion and the same physical growth maturation status were selected for the study. The subjects were divided among a Twin-block group (n = 25), a bionator group (n = 20), and a control group (n = 10). Pretreatment and posttreatment lateral cephalometric radiographs of the treatment group subjects, and prefollow-up and postfollow-up radiographs of the control group subjects, were traced manually and subjected to the pitchfork analysis. RESULTS: Statistical software was used for 1-way analysis of variance and multiple comparisons (post-hoc test, Bonferroni). A P value of .05 was considered statistically significant. Neither the Twin-block nor the bionator appliance significantly restricted forward growth of the maxilla (P = .476). Mandibular growth in the Twin-block subjects was significantly greater than in controls (P = .005). Mandibular growth was comparable in the control and the bionator subjects. Molar correction, overjet reduction, and proclination of the mandibular incisors were significantly greater (P = .000) in the treated subjects compared with the controls. CONCLUSIONS: Both the Twin-block and bionator appliances were effective in correcting molar relationships and reducing overjets in Class II Division 1 malocclusion subjects. However, the Twin-block was more efficient than the bionator in the treatment of Class II Division 1 malocclusion. PMID- 17110257 TI - Effects of mandibular symphyseal distraction osteogenesis on mandibular structures. AB - INTRODUCTION: The purpose of this study was to evaluate the effects of mandibular symphyseal distraction osteogenesis on dental and mandibular skeletal structures with a tooth- and bone-borne distraction device. METHODS: The sample comprised 20 patients, ranging in age from 15.8 to 23.3 years (mean, 20.1 +/- 2.3 years) at the start of treatment. The distraction device was activated 1 mm per day. The device was usually maintained in position about 90 days after surgery. Records were obtained at the start of treatment, at the end of distraction (11.4 +/- 2.2 days after surgery), and at follow-up periods (24.1 +/- 4.2 months after surgery). The records included posteroanterior cephalograms and study casts. The data were analyzed statistically by using the repeated measure analysis of variance and paired t test. RESULTS: Posteroanterior cephalometric analysis demonstrated no significant changes in bigonial widths and ramal angles at the end of distraction period. On the other hand, bimolar widths were significantly increased, whereas bicondylar widths were markedly decreased. The dental cast analysis indicated that the maximum amount of increase was found between the mandibular canines, and the widening effect gradually decreased from the mandibular canines to the second molars. The follow-up data confirmed that the treatment results were stable. CONCLUSIONS: The long-term findings indicate that mandibular symphyseal distraction osteogenesis is an efficient nonextraction treatment alternative for mandibular dental crowding to increase mandibular skeletal and dental arch widths. PMID- 17110258 TI - Assessment of cell proliferation during mandibular distraction osteogenesis in the maturing rat. AB - INTRODUCTION: The cellular mechanisms controlling distraction osteogenesis are not well understood. The purpose of this study was to examine the role of cell proliferation in the regulation of mandibular distraction osteogenesis. METHODS: Unilateral mandibular ramus osteotomies were performed on 125 3-month-old Sprague Dawley rats. The rats were randomized into 4 distraction rate groups and distracted for 5 days after 3-day latency. Rats (7 or 8 from each rate group) were killed at 4 time points. The rats received 5-bromo-2-deoxyuridine (BrdU) injections (40 mg per kilogram, i.p.) at day 3 (end of latency). RESULTS: Both intramembranous and endochondral ossification was seen in the osteogenesis area. BrdU+ mesenchymal progenitor cells were significantly higher at day 10 (P <.05) and were found most numerously around the sagittal middle portion of the gap (P <.01). The greatest numbers of BrdU+ osteocytes were seen at day 38 (P <.05). Both BrdU+ osteoclasts and chondrocytes peaked at day 24. CONCLUSIONS: Mesenchymal progenitor cells are mostly recruited in the early consolidation period, but they decrease in the middle and late consolidation periods during mandibular distraction osteogenesis. The rapid rate might suppress or sustain the proliferation and differentiation of mesenchymal progenitor cells during mandibular distraction osteogenesis. BrdU+ cells can survive throughout the entire experimental period of 5 weeks. PMID- 17110259 TI - Chronologic age and skeletal maturation of the cervical vertebrae and hand-wrist: is there a relationship? AB - INTRODUCTION: The aims of this study were (1) to investigate the relationship between chronologic age and maturation of cervical vertebrae, (2) to identify the relationship between chronologic age and maturation stage evaluated by hand-wrist radiographs, and (3) to determine whether the maturation of cervical vertebrae correlates with maturation indicated by hand-wrist radiographs in a Turkish population. METHODS: The samples were derived from lateral cephalometric and hand wrist radiographs of 503 subjects (213 male, 290 female; ages, 5.3-24.1 years). Cervical vertebral development was evaluated by the method of Hassel and Farman. Skeletal maturation of each hand-wrist radiograph was determined according to the method described by Bjork and Grave, and Brown's system. The Spearman rank-order correlation coefficients were estimated separately for males and females to measure the relationships among chronologic age, cervical vertebral maturation, and the skeletal maturation measured at the hand-wrist. RESULTS: The Spearman correlation coefficients were 0.72 (P <.001) between chronologic age and cervical vertebrae skeletal maturation, and 0.79 (P < .001) between chronologic age and maturation via hand-wrist radiographs. The correlation coefficient between hand wrist and cervical-vertebrae maturation was 0.86 (P <.001). CONCLUSIONS: The cervical-vertebrae maturation stages are clinically useful maturity indicators of the pubertal growth period Turkish subjects. PMID- 17110260 TI - Occlusal patterns in patients with idiopathic scoliosis. AB - INTRODUCTION: Idiopathic scoliosis is an orthopedic condition characterized by faulty posture. It might also be associated with some mild forms of facial asymmetry or dental deviations. The aim of the study was to clinically examine the occlusions of patients with idiopathic scoliosis. METHODS: Ninety-six consecutive orthopedic patients with idiopathic scoliosis were examined. The orthopedic data of this group were recorded from their hospital files. The occlusal features of a random group of 705 Ashkenazi children served as the control. Frequency distributions were compared with the chi-square test. RESULTS: The distribution of the Angle classes of malocclusion was significantly different in the 2 groups (P = .0001) because of many Class II subdivision patients in the orthopedic group. Other evidence of asymmetrical malocclusion was found in upper (P =.002) and lower midline deviations (P =.0001), and a higher frequency of anterior (P = .024) and posterior (P =.020) crossbites. In the experimental group, no association was found between site, side, or severity of scoliosis and the appearance or site of the malocclusion features examined. CONCLUSIONS: Patients with idiopathic scoliosis have asymmetric features of malocclusion compared with a random population. PMID- 17110261 TI - Effects of early loss of permanent first molars on the development of third molars. AB - INTRODUCTION: The aim of this study was to evaluate whether early loss of the permanent first molars has an effect on the developmental stages and eruptive conditions of the third molars. METHODS: Panoramic radiographs and dental casts of 165 adolescent patients (103 boys, 62 girls) with unilateral early loss of a permanent first molar were used to assess the developmental stages of the third molars. The formation stages of third molars on the extraction sides were evaluated, and the formative conditions of the contralateral teeth were used as controls. The Mann-Whitney U test was used to assess sex differences in formation stages of the third molars. The Kruskal-Wallis analysis of variance (ANOVA) was used to determine whether there were significant differences in the developmental stages of the third molars between the various extraction quadrants. In addition, the Wilcoxon signed rank test and the paired t test were used to compare the differences in the developmental stages and eruptive conditions of the third molars between the extraction and control sides. RESULTS: The Mann-Whitney U test showed no significant sex differences for the formation stage of the third molars. Therefore, the data from both sexes were pooled. No statistically significant differences were found in the developmental stages of the third molars between the various extraction quadrants in the same jaw and between those in the mandible and maxilla. However, statistically significant differences were found in developmental stages (P <.001) and eruptive conditions (P <.05) of the third molars between the extraction and control sides. The development of the third molars on the extraction side was significantly accelerated compared with the contralateral teeth. CONCLUSIONS: Early loss of the permanent first molars might have an accelerating effect on the development of the third molar on the extraction side compared with the contralateral teeth. Therefore, emergence of the third molars on the extraction side might be hastened, and these teeth might erupt earlier than the contralateral teeth. PMID- 17110262 TI - Physical properties of root cementum: part 8. Volumetric analysis of root resorption craters after application of controlled intrusive light and heavy orthodontic forces: a microcomputed tomography scan study. AB - INTRODUCTION: Intrusion is a critical type of orthodontic tooth movement in relation to external root resorption. Our aims in this prospective randomized clinical trial were to quantify, 3 dimensionally, the amount of root resorption when controlled light and heavy intrusive forces were applied to human premolars and to establish the sites where root resorption is more prevalent. METHODS: Fifty-four maxillary first premolars, orthodontically indicated for extraction from 27 patients (left and right maxillary first premolars from each), were intruded for 28 days with buccal and palatal beta-titanium-molybdenum alloy 0.017 x 0.025-in cantilever springs. The patients were randomly divided into 3 groups, and various levels of force were used: group 1, heavy force (225 g) on 1 side and control force (0 g) on the contralateral side; group 2, light force (25 g) on 1 side and control force (0 g) on the contralateral side; group 3, light force (25 g) on 1 side and heavy force (225 g) on the contralateral side. After the experimental period, the teeth were extracted under a strict protocol to prevent root surface damage and analyzed by using a microcomputed tomography scan x-ray system (SkyScan-1072, Skyscan, Aartselaar, Belgium) and specially designed software for direct volumetric measurements. RESULTS: The volume of the root resorption craters after intrusion was found to be directly proportional to the magnitude of the intrusive force applied. The results showed that the control group had fewer and smaller root resorption craters, the light force group had more and larger root resorption craters than the control group, and the heavy force group had the most and the largest root resorption craters of all groups. A trend of linear increase in the volume of the root resorption craters was observed from control to light to heavy groups, and these differences were statistically significant. The mean volumes of the resorption craters in the light and heavy force groups were 2 and 4 times greater than in the control groups, respectively. The mesial and distal surfaces had the greatest resorption volume, with no statistically significant difference between the 2 surfaces. PMID- 17110263 TI - Assessing treatment outcomes for a graduate orthodontics program: follow-up study for the classes of 2001-2003. AB - INTRODUCTION: Based on an initial 3-year study (1998-2000) of clinical outcomes, specific changes were made in the clinical protocol in the orthodontic clinic at Indiana University, Indianapolis, Ind. To evaluate these adjustments, a follow-up study with the same methods assessed the clinical outcomes for the next 3 years (2001-2003). METHODS: The 3 changes introduced in 2000 were assessment of prefinish records, education of residents about previous outcomes, and more frequent practice evaluations by the program director. The American Board of Orthodontics (ABO) objective grading system (OGS) and a supplemental comprehensive clinical assessment (CCA) were used to evaluate the results for all patients completed by the classes of 2001 through 2003 (n = 437). RESULTS: The mean ABO OGS, CCA, and combined scores were 25.19 +/- 11.16, 4.38 +/- 2.74, and 29.57 +/- 13.01, respectively. A progressive improvement was noted from 2001 to 2003. The ABO OGS and the combined scores were significantly (P <.001) improved in 2001 through 2003 compared with 1998 through 2000. The CCA score tended to improve, but it was not significantly different. Significant improvements were seen for maxillary and mandibular first order, mandibular second order, mandibular third order, overjet, occlusal contacts, occlusal relationships, and root angulations (P <.001). As with the previous study, excessive treatment times correlated with worse clinical outcomes. CONCLUSIONS: Changes implemented by the graduate program were effective for improving treatment outcomes. Cast scores were more amenable to improvement than other more comprehensive outcomes. PMID- 17110264 TI - A report of the ABO Resident Clinical Outcome Study (the pilot study). AB - INTRODUCTION: A 4-year collaborative project between the American Board of Orthodontics (ABO) and 15 American orthodontic graduate programs concluded at the ABO Clinical Examination in February 2006. METHODS: Fifty recent graduates (the pilot study group) presented cases that were treated in their graduate programs as partial fulfillment of the requirements for ABO certification. The examinees were tested by calibrated ABO examiners and required to satisfy traditional ABO standards. They presented appropriate case reports that contained the ABO's 3 measurement instruments. Thirty-nine practicing orthodontists (the control group) presented cases according to the traditional ABO Clinical Examination process. RESULTS: Ninety percent of the pilot study group and 85% of the control group successfully accomplished ABO certification. There was a difference of 2.38 points between the 2 groups for the mean total scores of the cases that passed. The pilot study group presented cases that met the historic averages for case complexity (discrepancy index). CONCLUSIONS: Residents in orthodontic programs are able to treat cases to ABO standards of quality. PMID- 17110265 TI - ABO initial certification examination: official announcement of criteria. PMID- 17110266 TI - A comparison of different ligation methods on friction. AB - INTRODUCTION: An elastomeric module with a polymeric coating has been developed to reduce the friction of sliding mechanics. This in-vitro study examined the stability of the coating and compared the frictional properties of coated modules with those of other common ligation methods. METHODS: Six ligation methods (regular uncoated, slick [coated], conventional silver, easy-to-tie, silicone impregnated, and standard silver modules) were used with standard stainless steel brackets and 0.019 x 0.025-in archwires, and resistance to movement was measured. Two self-ligating (Speed [Strite Industries, Cambridge, Ontario, Canada] and Damon 2 [Sybron Dental Specialities Ormco, Orange, Calif]) brackets were also tested. RESULTS: The Damon 2 self-ligating brackets produced less friction than the other ligation methods, followed by the coated modules. There was no significant difference between the frictional resistance of brackets ligated with regular uncoated, silicone-impregnated, and easy-to-tie modules. Speed self ligating brackets produced less friction than regular uncoated, conventional silver, and standard silver modules. The frictional properties of coated modules were not significantly affected by repeating the test 5 times or by storage in saliva for a week. CONCLUSIONS: Damon 2 brackets produced no recordable friction of ligation. Coated modules produced 50% less friction than all other ligation methods except Damon 2. The coating was resistant to the simulated effects of the oral environment. Different methods of human saliva application were found to affect the frictional properties of the coating. PMID- 17110267 TI - Upper midline correction in conjunction with rapid maxillary expansion. AB - INTRODUCTION: The purpose of this study was to investigate a simple method for correcting upper dental midline shift, by physiologic movement of the incisors during the retention period of rapid maxillary expansion. METHODS: Thirty-two patients with upper midline shifts due to constricted maxillae and anterior crowding were selected. All had undergone radiological and clinical examinations to be sure that the midline shift was not functional and was caused only by crowding. The patients were randomly divided into 2 groups. In both groups, acrylic cap splint type expansion appliances were used. In group 1, the acrylic cap covered all the dentition except the incisors and the canine on the shifted side. After expansion, the incisors on the shifted side were allowed to move toward the midline, and the incisors of the other side were held by the acrylic cap. In group 2, the acrylic cap covered only the posterior teeth, from the first premolars on both sides. After expansion, the incisors on the shifted side were allowed to move toward the midline diastema, whereas those on the other side were held in place by ligation of the brackets, which had been placed immediately after expansion. RESULTS: In both groups, the midline shift was corrected by the movement of the shifted incisors toward the midline diastema, without orthodontic force. CONCLUSIONS: Residual or unwanted forces produced during rapid maxillary expansion can be used to correct an upper dental midline shift. PMID- 17110268 TI - Correction of deep overbite and gummy smile by using a mini-implant with a segmented wire in a growing Class II Division 2 patient. AB - A boy, aged 10.5 years, with a Class II molar relationship and a very deep overbite, complaining of a gummy smile and anterior crowding, was treated nonextraction with a mini-implant and Twin-block and edgewise fixed appliances. Severely extruded and retroclined maxillary incisors were intruded and proclined with a nickel-titanium closed-coil spring anchored to a mini-implant and segmented wires; this resolved the gummy smile and deep overbite efficiently without extruding the maxillary molars or opening the mandible. The mandibular incisors were proclined without direct orthodontic force during intrusion of the maxillary incisors; this helped the nonextraction treatment of mandibular incisor crowding. The Twin-block appliance with high-pull headgear promoted mandibular growth, restrained maxillary growth, and changed the canine and molar relationship from Class II to Class I. The patient's overbite and overjet were overtreated, and, 1 year postretention, the patient maintained a good overbite and overjet. PMID- 17110269 TI - Litigation, legislation, and ethics. The role of expert witnesses in claims for lack of informed consent. PMID- 17110270 TI - Ending the isolation of depression from neurology practice. PMID- 17110271 TI - Neuroprotection in Parkinson's disease: and now for something completely different? PMID- 17110272 TI - Predicting the course of Guillain-Barre syndrome. PMID- 17110273 TI - Nimodipine for traumatic [corrected] subarachnoid haemorrhage: the end of the road or better trials? PMID- 17110274 TI - NICE and anti-dementia drugs: a triumph of health economics over clinical wisdom? PMID- 17110275 TI - NSA guidelines for management of transient ischaemic attacks. PMID- 17110276 TI - RNAi gets its prize. PMID- 17110277 TI - The long and short of it: telomeres and the brain. PMID- 17110278 TI - Plasma amyloid-beta concentrations in Alzheimer's disease: an alternative hypothesis. PMID- 17110279 TI - Plasma amyloid-beta concentrations in Alzheimer's disease: an alternative hypothesis. PMID- 17110281 TI - TCH346 as a neuroprotective drug in Parkinson's disease: a double-blind, randomised, controlled trial. AB - BACKGROUND: There is an important unmet medical need in Parkinson's disease for a neuroprotective treatment that slows or stops disease progression. TCH346 is a potent anti-apoptotic drug that protects against loss of dopaminergic neurons in laboratory models. Our aim was to assess TCH346 as a neuroprotective drug in patients with Parkinson's disease. METHODS: Patients presenting at 45 international movement disorder clinics with early untreated Parkinson's disease were assessed as part of this parallel-group, double-blind, randomised controlled trial. 301 eligible patients were randomly assigned 12-18 months' treatment with TCH346 at a daily dose of 0.5 mg (n=78), 2.5 mg (n=79), or 10 mg (n=73), or placebo (n=71), followed by a 4 week washout period. The primary outcome measure was time to development of a disability requiring dopaminergic treatment. Secondary outcome measures were the annual rate of change in the unified Parkinson's disease rating scale (UPDRS) and the PDQ-39, a measure of quality of life. Analyses were by intention-to-treat. This study is pending registration with . FINDINGS: 255 patients completed the study. TCH346 did not differ from placebo for any of the study outcomes. Treatment was needed in 26 (34%) patients in the TCH346 0.5 mg group, 30 (38%) in the TCH346 2.5 mg group, 24 (33%) in the TCH346 10 mg group, and 23 (32%) in the placebo group. There were no significant differences between groups. There were no differences between groups in the annual change in the UPDRS or PDQ-39 either. Few patients withdrew because of adverse events and none was judged to be related to the study intervention. INTERPRETATION: TCH346 did not show evidence of a neuroprotective effect. The discrepancy between the preclinical promise of TCH346 and the clinical outcome could have arisen because of the use of laboratory models that do not accurately reflect the pathogenesis of Parkinson's disease, the doses of study drug used, insensitive clinical endpoints, and the patient population selected for study. PMID- 17110282 TI - Clinical and electrophysiological predictors of respiratory failure in Guillain Barre syndrome: a prospective study. AB - BACKGROUND: Respiratory failure is the most serious short-term complication of Guillain-Barre syndrome and can require invasive mechanical ventilation in 20-30% of patients. We sought to identify clinical and electrophysiological predictors of respiratory failure in the disease. METHODS: We prospectively assessed electrophysiological data and clinical factors, including identified predictors of delay between disease onset and admission, inability to lift head, and vital capacity, in patients admitted with Guillain-Barre syndrome. We related these factors to subsequent need for ventilatory support. Neurophysiological findings were classified as demyelinating, axonal, equivocal, unexcitable, or normal. Predictive values of clinical and electrophysiological data were tested using classification trees to build up a predictive model. This model was initially built up in a two-third (fitting set) then validated in a one-third (validation set) of the total sample. The fitting and validation sets were randomly selected. We also assessed the predictive value of this model for disability at 6 months. FINDINGS: From 1998, to 2006, 154 patients with Guillain-Barre syndrome were included in the study and 34 (22%) were subsequently ventilated. Demyelinating Guillain-Barre syndrome was more common in patients who went on to be ventilated than in those who were not (85%vs 51%, p=0.0003). Vital capacity and the proximal/distal compound muscular amplitude potential (p/dCMAP) ratio of the common peroneal nerve were retained in the tree model, with a probability of needing ventilation of less than 2.5% in patients with a ratio of greater than 55.6% and a vital capacity more than 81% of predicted. A p/dCMAP ratio of the peroneal nerve less than 55.6% and age older than 40 years were retained as independent predictors of disability at 6 months. INTERPRETATION: Neurophysiological testing is helpful for assessing risk of respiratory failure, which is highest in patients with evidence of demyelination and very low in those without both 55.6% conduction block of the common peroneal nerve and a 20% reduction in vital capacity. PMID- 17110283 TI - Effect of nimodipine on outcome in patients with traumatic subarachnoid haemorrhage: a systematic review. AB - BACKGROUND: Despite several randomised controlled trials, there is still much debate whether nimodipine improves outcome in patients with traumatic subarachnoid haemorrhage. A 2003 Cochrane review reported improved outcome with nimodipine in these patients; however, because the results of Head Injury Trial (HIT) 4 were only partly presented there is still discussion whether patients with traumatic subarachnoid haemorrhage should be treated with this drug. Here, we present data from all head-injury trials, including previously unpublished results from HIT 4. METHODS: We systematically searched PubMed and EMBASE databases using the following combinations of variables: "nimodipine" or "calcium antagonist" with "traumatic subarachnoid haemorrhage", "head injury", "head trauma", "brain injury", or "brain trauma". Bayer AG and all principal investigators or corresponding authors of the identified studies were contacted for additional information. FINDINGS: Five manuscripts were identified, describing the results of four trials. We obtained additional data from HIT 1, 2, and 4. In total, 1074 patients with traumatic subarachnoid haemorrhage were included. The occurrence of poor outcome was similar in patients treated with nimodipine (39%) and those treated with placebo (40%); odds ratio was 0.88 (95% CI 0.51-1.54). Mortality rates did not differ between nimodipine (26%) and placebo (27%) treated patients (odds ratio 0.95; 95% CI 0.71-1.26). INTERPRETATION: Our results do not lend support to the finding of a beneficial effect of nimodipine on outcome in patients with traumatic subarachnoid haemorrhage as reported in an earlier Cochrane review. PMID- 17110284 TI - Brain imaging of clinical pain states: a critical review and strategies for future studies. AB - Research into brain imaging of pain is largely dominated by experimental acute pain studies. Applied study paradigms have evolved a lot over past years and the ensuing results have furthered enormously our understanding of acute-pain processing. In sharp contrast, published work on brain-imaging in chronic pain remains scant. Furthermore, the results of these studies are highly incongruent, which could be explained by the fact that patient populations studied varied largely in terms of pain history, pain distribution, cause of pain, and psychological set-up. To circumvent these problems, several investigators have used surrogate models of neuropathic pain, but the validity of these models is highly questionable. In this Review we critically discuss the problems and shortcomings of most published reports on chronic pain and we propose some strategies for future studies. We argue that the post-operative pain model is highly appealing since it opens perspectives for prospective longitudinal studies with repeated assessments and it enables control for many confounding factors, which hamper the interpretation of most current studies. We also plead for a multimodal imaging approach in which classic brain-activation studies are supplemented with genetic, neurochemistry, brain morphometry, and transcranial magnetic stimulation studies. PMID- 17110285 TI - Histological classification and molecular genetics of meningiomas. AB - Meningiomas account for up to 30% of all primary intracranial tumours. They are histologically classified according to the World Health Organization (WHO) classification of tumours of the nervous system. Most meningiomas are benign lesions of WHO grade I, whereas some meningioma variants correspond with WHO grades II and III and are associated with a higher risk of recurrence and shorter survival times. Mutations in the NF2 gene and loss of chromosome 22q are the most common genetic alterations associated with the initiation of meningiomas. With increase in tumour grade, additional progression-associated molecular aberrations can be found; however, most of the relevant genes are yet to be identified. High throughput techniques of global genome and transcriptome analyses and new meningioma models provide increasing insight into meningioma biology and will help to identify common pathogenic pathways that may be targeted by new therapeutic approaches. PMID- 17110286 TI - Effects of seizures on developmental processes in the immature brain. AB - Infants and children are at a high risk for seizures compared with adults. Although most seizures in children are benign and result in no long-term consequences, increasing experimental animal data strongly suggest that frequent or prolonged seizures in the developing brain result in long-lasting sequelae. Such seizures may intervene with developmental programmes and lead to inadequate construction of cortical networks rather than induction of neuronal cell loss. As a consequence, the deleterious actions of seizures are strongly age dependent: seizures have different effects on immature or migrating neurons endowed with few synapses and more developed neurons that express hundreds of functional synapses. This differential effect is even more important in human beings and subhuman primates who have an extended brain development period. Seizures also beget seizures during maturation and result in a replay of development programmes, which suggests that epileptogenesis recapitulates ontogenesis. Therefore, to understand seizures and their consequences in the developing brain, it is essential to determine how neuronal activity modulates the main steps of cortical formation. In this Review, we present basic developmental principles obtained from animal studies and examine the long-lasting consequences of epilepsy. PMID- 17110287 TI - Antiepileptic drugs in development. AB - BACKGROUND: Despite the success of several new antiepileptic drugs, about one third of patients with epilepsy are not seizure free on medication. Improvement in this situation might lie in drugs that are currently in development. RECENT DEVELOPMENTS: Some new antiepileptic drugs are modifications of those already available, referred to in this Rapid Review as evolutionary drugs. These modifications of existing drugs are developed to improve effectiveness, often by increasing tolerability. Other drugs work by new mechanisms and are usually discovered through screening of animal models. WHERE NEXT? The large number of drugs currently in clinical trials provides a measure of hope for patients whose epilepsy is not controlled with currently available medication. In the future, this range of antiepileptic drugs will probably increase because of the use of new animal models, discovery of new basic mechanisms of epileptogenesis, acceleration of proof of principle studies in people, and development of new methods of drug delivery. PMID- 17110288 TI - The HTLV-1 neurological complex. AB - Human T-cell lymphotropic virus type 1 (HTLV-1) is a retrovirus that infects about 20 million people worldwide and causes immune-mediated diseases of the nervous system. The classic neurological presentation of HTLV-1 infection is a myelopathy called HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). However, HAM/TSP is not the only neurological outcome that can result from HTLV-1 infection. In this Personal View, we show that HTLV-1 has a broader neurological spectrum than the names HAM/TSP suggest and that people infected with this virus can present with various isolated or assorted syndromes. PMID- 17110290 TI - Exposure to airborne particulate matter in the ambient, indoor, and occupational environments. AB - Exposure to airborne particulate matter results in various adverse health effects. Unlike other pollutants, such as ozone, sulfur dioxide, carbon monoxide, and oxides of nitrogen, for which there is significant exposure, particulate matter exposure is much more complex because it is not a single chemical species or even a limited number of chemical species. Particulate matter includes various chemical species in particles having a wide range of diameters and shapes that have widely varying toxicities. People are exposed to particles in the ambient environment, in indoor spaces, and in the occupational environment. This article reviews the information available on the concentrations of particulate matter and its composition in these general environmental categories. PMID- 17110291 TI - Aerosol dosimetry considerations. AB - The concept of dose is fundamental to the discipline of toxicology. For inhaled particles, dose considerations include the sequential processes of inhalation, particle deposition, and particle clearance. Several important parameters modify each of these processes, including environmental, anatomic, and physiologic factors. When such factors are considered, it is possible to identify subpopulations and individuals who are likely to receive particle doses that greatly exceed those for the average population. Higher than average doses can be expected for people who are young, have certain acute or chronic lung diseases, are engaged in exercise, or are exposed in close proximity to sources of air pollutants. Although considerable research has improved the understanding of inhaled particle doses, much is still to be learned before high-risk groups and individuals can be protected properly. PMID- 17110292 TI - Translocation and effects of ultrafine particles outside of the lung. AB - Ultrafine, or nano-, particles (< 100 nm) have been associated in epidemiological, human clinical, and animal studies with adverse cardiopulmonary outcomes. Deposition of inhaled ultrafine particles in the respiratory tract is mainly governed by diffusion and is most efficient for alveolar regions of the lung, although deposition occurs in other regions, too. The nose is also a very efficient filter for smaller ultrafine (< 5 nm, diffusion) particles. Solid poorly-soluble ultrafine particles are not efficiently cleared via mucociliary or macrophage-mediated mechanisms and are, thus, likely to be taken up by epithelial cells and translocate to extrapulmonary sites (interstitium, lymph and blood circulation, neurons). These translocation processes are explored here as well potential consequences that result from exposure of extrapulmonary organs to inhaled ultrafine particles. PMID- 17110293 TI - Inflammation and airborne particles. AB - Inflammation provides a potential mechanistic link between inhalation of particles and the diverse health effects found in epidemiologic studies. Considerable uncertainty remains as to the importance of the inflammation in mediating these effects and where that inflammation is occurring: lung, vascular endothelium, or distant organs, including the heart. This article briefly reviews the role of inflammation in pulmonary and cardiovascular disease and explores the evidence that the health effects of PM exposure are mediated, at least in part, by inflammation. PMID- 17110294 TI - The role of reactive oxygen species and oxidative stress in mediating particulate matter injury. AB - Numerous reports link oxidative stress to particulate matter (PM)-induced adverse health effects. Increasing evidence is being collected that reactive oxygen species and oxidative stress are involved in PM-mediated injury. The physical characteristics and the chemical composition of PM play a key role in reactive oxygen species generation in vitro and in vivo. According to the hierarchical oxidative stress hypothesis, antioxidant phase II enzymes protect against PM induced inflammation and cytotoxicity. This concept is useful in understanding PM induced disease models, susceptibility, and biomaker development to access exposures outcomes and is useful for developing therapeutic intervention in PM induced adverse health effects. PMID- 17110295 TI - Long-term effects of exposure to particulate air pollution. AB - Considerable work has been done to elucidate the effects of polluted air, most of which has studied acute effects of particles. Studies suggest that the effects of longer term exposures are more than just the daily sum of the acute effects. Because most of the studies of acute effects have examined changes in health status occurring within days of the exposure, this article takes a broad definition of long-term exposure to include averaging times of months to years. It concludes that health effects increase as length of exposure increases, but much of that increase occurs within the first year. PMID- 17110296 TI - Responses of the heart to ambient particle inhalation. AB - This article focuses on responses to ambient particles by the heart. Available data from human studies and animal studies are reviewed in an attempt to find a common understanding in the findings. The pathophysiologic mechanisms responsible for these health effects are likely to be complex, and it is highly probable that several different mechanisms work in concert. Current evidence suggests that inhaled particles exert their effects on the heart via the autonomic nervous system and via the coronary vasculature. Direct effects on the myocardium by inhaled ambient particles or their constituents require more research. PMID- 17110297 TI - Effects of particulate air pollution on hemostasis. AB - Exposure to particulate air pollution is associated with acute and chronic cardiovascular morbidity and mortality. The mechanisms involved in these effects are not fully elucidated. Research has proved that fine particles, principally the ultrafine fraction, which are predominantly derived from combustion of fossil fuel, are the most toxic. Recent clinical and experimental studies have reported mechanistic observations linking fine and ultrafine particles to the coagulation cascade, platelet function, and subsequent development of atherosclerosis and thrombosis. These effects have been explained either by release of soluble mediators by the lungs, which affect blood coagulation parameters, or by the direct translocation of ultrafine particles into the systemic circulation or the alteration of autonomic cardiac control. Despite recent advances, additional studies are needed to investigate the pathophysiologic mechanisms linking particulate air pollution and hemostasis. PMID- 17110298 TI - Implications for occupational exposure to particulate matter. AB - The demonstrated effects of lower levels of ambient particles on cardiovascular and respiratory system morbidity and mortality were initially surprising in light of current concepts of occupational particle exposure and acute and chronic cardiopulmonary effects. Specifically, the exposure levels, as defined by the weight of the particles per liter of breathing air, at which recognized disease occurs under workplace conditions are considerably higher than the observed levels of ambient particles associated with serious adverse health effects. The possible reasons for this difference have not been adequately addressed. To further address this question, a re-examination of workplace exposure-response relationships is needed, which may include emphasis on measuring exposures to fine and ultrafine particles rather than to total particle mass concentration alone. PMID- 17110299 TI - Surveillance of dysplasia in inflammatory bowel disease: The gastroenterologist pathologist partnership. AB - Cancer prevention in inflammatory bowel disease depends on the detection of precancerous dysplasia during scheduled screening and surveillance colonoscopy, but the detection and diagnosis of dysplasia remain challenging. In this article, we review the risks of cancer and dysplasia in ulcerative colitis and the current prevention recommendations, and through a sample case, we demonstrate an approach that involves an active partnership between the gastroenterologist or surgeon and pathologist. We address the challenge of management of polypoid lesions and incorporate new information about degree of inflammation as an additional risk of neoplasia in these patients. PMID- 17110300 TI - Portal hypertension and hepatocellular carcinoma: prognosis and beyond. PMID- 17110301 TI - Treatment of clonal refractory celiac disease or cryptic intraepithelial lymphoma: A long road from bench to bedside. PMID- 17110302 TI - Best practices: academic gastroenterology practices. PMID- 17110303 TI - Low-grade dysplasia in Barrett's esophagus. PMID- 17110305 TI - The long-term efficacy of pneumatic dilation and Heller myotomy for the treatment of achalasia. PMID- 17110308 TI - Undifferentiated connective tissue diseases (UCTD). AB - The term undifferentiated connective tissue diseases is used to define conditions characterized by the presence of signs and symptoms suggestive of a systemic autoimmune disease that do not satisfy the classificative criteria for defined connective tissue diseases (CTD) such as systemic lupus erythematosus (SLE), Sjogren's syndrome (SS), rheumatoid arthritis (RA) and others. A small percentage of patients presenting with an undifferentiated profile will develop during the first year follow up of a full blown CTD, however an average of 75% will maintain an undifferentiated clinical course. These patients may be defined as having a stable undifferentiated connective tissue diseases (UCTD). The most characteristic symptoms of UCTD are represented by arthritis and arthralgias, Raynaud's phenomenon, leukopenia, while neurological and kidney involvement are virtually absent. Eighty percent of these patients have a single autoantibody specificity, more frequently anti-Ro and anti-RNP antibodies. Stable UCTD are considered as distinct clinical entities and therefore it has been proposed to define those conditions as UCTD. Classificative criteria have also been proposed and a work to better define them is still under way. PMID- 17110309 TI - Possible pathogenic nature of the recently discovered TT virus: does it play a role in autoimmune rheumatic diseases? AB - Pathogenesis of viral origin has long been suggested in autoimmune rheumatic diseases. Beside the well-defined virus induced transient or chronic rheumatic diseases often resembling systemic autoimmune disorders such as rheumatoid arthritis, viruses can contribute to disease pathogenesis by several different pathomechanisms. TT virus is a recently discovered virus of extremely high genetic diversity which commonly infects humans. Despite accumulated evidence on the biological characteristics of TTV, its pathogenicity is still in question; many consider TTV as a harmless endosymbiont. The recent paper overviews the biology of TT virus and investigates the hypothesis that TTV might have a causative role in human diseases with special attention to the possibility that TTV might trigger autoimmunity in rheumatic disorders. PMID- 17110310 TI - Interaction of giant phospholipid vesicles containing cardiolipin and cholesterol with beta2-glycoprotein-I and anti-beta2-glycoprotein-I antibodies. AB - Antiphospholipid syndrome is characterized with thrombotic events and/or pregnancy morbidity and antiphospholipid antibodies (aPL). The most common antigen for aPL is beta2-glycoprotein-I (beta(2)GPI), a plasma protein binding to negatively charged phospholipids. The influence of aPL on coagulation is not well understood. Giant phospholipid vesicles (GPVs) are a convenient in vitro system for studying interactions between phospholipid membranes and proteins resulting in the change of the vesicles' configuration. We aimed to set up an in vitro model and to study changes in the morphology of GPVs with high content of cardiolipin upon addition of beta(2)GPI and/or IgG fraction of a patient with antiphospholipid syndrome (APS). Addition of the IgG fraction of the APS patient caused lateral segregation of the membrane inclusions and adhesion of GPVs. Addition of beta(2)GPI caused adhesion of GPVs. Addition of both, the patient IgG fraction and beta(2)GPI caused adhesion of vesicles to the glass slides and to each other, formation of pores and burst of vesicles. Our results indicate that adhesion of the cardiolipin-containing vesicles does not seem specific for added proteins, rather, it indicates electrostatic and curvature-mediated interactions between the membrane constituents. PMID- 17110311 TI - Immunohistopathology of Sjogren's syndrome. AB - Sjogren's syndrome (SS) is characterized by keratoconjunctivitis sicca and xerostomia, which occur in an autoimmune lacrimal and salivary gland disease characterized by lymphocyte infiltrates of exocrine glands and/or Sjogren's syndrome autoantibody production. It has been reported that aquaporin-5 distribution is abnormal in SS, perhaps as a result of paracrine effect of TNF alpha. Also the neurogenic regulation of the salivary gland is impaired in SS. Apart from functional changes, the syndrome is also characterized by structural abnormalities of the secretory acinar apparatus. The acinar basement membrane is abnormal as it lacks laminin alpha1 chain, which may impair its capability to induce the progenitor cells to differentiate to acinar cells. CRISP-3 and TMPRSS 2 can be used as androgen markers and LIV-1 and Cyr61 as estrogen markers to study the sexual dimorphism of the salivary glands. Patients with SS seem to have low concentrations of dehydroepiandrosterone, which may predispose women and the exocrine glands to this syndrome. PMID- 17110312 TI - Serum amyloid A in autoimmune thrombosis. AB - The objectives of this study were (1) to determine how levels of serum amyloid A (SAA), high sensitivity C-reactive protein (CRP) and interleukin-6 (IL-6) correlate to autoimmune diseases in patients with or without thrombosis, and (2) to discuss the parameters that influence the relative SAA values. SAA, CRP and IL 6 concentrations were determined by enzyme linked immunosorbent assay (ELISA). 84 patients with secondary antiphospholipid syndrome (SAPS), primary antiphospholipid syndrome (PAPS), systemic lupus erythematosus with antiphospholipid antibodies (SLE+aPL), SLE, venous thrombosis (VT), arterial thrombosis (AT) were compared to healthy donors (n=60). The percentages of patients above cut-off were highest in the SAPS, SLE and SLE+aPL groups. Significant differences were observed between healthy donors and inflammatory groups of patients (SAPS and SLE+aPL) in all three measured parameters. SAA and CRP were shown to be correlated to a greater extent in SAPS patients than SLE+aPL patients. In summary, this cross-sectional, retrospective, small study and accompanying clinical considerations limit the ability to make definite conclusions. SAA would not serve as a useful marker for venous, arterial thrombosis or PAPS (pro-coagulant events). It could however, be a good predictor of progression from a non-inflammatory thrombotic condition to an inflammatory one. PMID- 17110313 TI - Changes in avidity and specificity of IgG during electro-oxidation. Relevance of binding of antibodies to beta2-GPI. AB - The immune response may be changed due to altered proteins or modifications of immunoglobulins, including oxidative processes. The susceptibility to oxidative modifications depends greatly on amino-acid moiety composition due to chemical characteristics (instability) of their side-chains. Initial steps of oxidation may change the specificity and avidity of immunoglobulins due to chemical alteration of the hypervariable region. The oxidation of antibodies increases the hydrophilic nature of the paratopes and makes them more susceptible for the binding to cationic surfaces even without the strong surface-to-surface fitting. The electro-oxidation of IgG significantly changes the immunoreactivity and specificity of IgG fractions, regardless of the initial immunoreactivity to a specific autoantigen also in healthy persons. Data are presented on changes in the immunoreactivity as well as the avidity of antibodies against beta2 glycoprotein I after being exposed to direct current. ELISA measurements showed increased reactivity of anti-beta2-glycoprotein I antibodies at the beginning and various, fluctuating results after prolonged exposure to electro-oxidation. Inter individual differences in chemical stability of immunoglobulins and patient's antioxidative status may influence the range of their alterations and their impact on health/disease balance. PMID- 17110314 TI - Rheumatic diseases and Klinefelter's syndrome. AB - The Klinefelter's syndrome (KS) is not a rare gonosomal aberration occurring in males. The disorder is characterized by microorchidism. Another typical although not constant symptom of this disorder is gynecomastia with almost normal male secondary sex characteristics. The etiology of the disease remains unexplained. Previous studies have shown that this disorder is a genetic chromosomal abnormality associated with the presence of one additional chromosome due to abnormal division. Thus, the affected individual has 47 chromosomes with the resulting chromosomal constellation of XXY (classical form) or 46,XY/47,XXX (mosaic form). Large population studies estimate the incidence of KS at 1:1000 live born male babies [Hammerton JL, Canning N, Ray M, et al. A cytogenic survey of 14,069 newborn infants. I. Incidence of chromosome abnormalities. Clin Genet 1975;8:223-243]. The locomotor apparatus of persons affected by the syndrome is characterized by acromicria, clinodactyly, concrescence of thoracal vertebral bodies and spinal osteoporosis in not only individuals of older age but also in younger persons. In 1960s and 1970s, reports were published on the concurrence of the KS with autoimmune diseases. The aim of our article is to discuss case reports on the KS published by authors from our institute as well as to present an overview of the reports published so far, mainly abroad. PMID- 17110315 TI - Autoantibodies to citrullinated antigens in (early) rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a common, systemic autoimmune disease characterized by chronic inflammation of the synovium, that can lead to progressive joint destruction and in many cases results in severe disability and poor quality of life. With the availability of more sophisticated and effective therapies and with increasing evidence that the first few months of disease represent an unique therapeutic opportunity and that such early therapeutic intervention is crucial in preventing irreversible joint damage, it is widely accepted that early and accurate diagnosis of RA is critical in disease management. Within the last three years a growing number of publications have reported that the second generation anti-CCP (cyclic citrullinated peptide) test may become the marker of choice for diagnosing early RA as it appears to be highly specific for the disease with a sensitivity comparable to the widely used but less specific rheumatoid factor test. Additionally, anti-CCP2 positivity can predict future development of RA in both asymptomatic individuals and in patients with undifferentiated arthritis. Furthermore, antibody levels at presentation can correlate with progression to erosive disease. PMID- 17110316 TI - Malignancies and soluble tumor antigens in rheumatic diseases. AB - Paraneoplastic symptoms, caused by a malignancy, but not directly related to invasion by the tumor or its metastases are the result of a wide variety of tumor derived biologic mediators like hormones, peptides, antibodies, cytotoxic lymphocytes, autocrine and paracrine mediators. Recognition of paraneoplastic syndromes is important, as it may lead to an early diagnosis of cancer. There is some evidence that systemic inflammatory diseases, such as rheumatoid arthritis (RA), lupus, scleroderma or dermatomyositis may increase the risk for the development of malignancies, predominantly lymphoproliferative disorders. However, reports are somewhat controversial. Immunosuppressive and cytotoxic drugs used in antirheumatic therapy, such as methotrexate, cyclophosphamide, azathioprine or anti-TNF biologicals may also lead to the development of such tumors. Tumor-associated antigens may be produced by inflammatory cells and their production may be increased in RA and other autoimmune diseases. PMID- 17110317 TI - Insulin resistance, chronic inflammatory state and the link with systemic lupus erythematosus-related coronary disease. AB - The association of SLE with atherosclerosis suggests a common pathogenic mechanism. SLE and atherosclerosis are immune complex-mediated diseases. The integration of metabolism and immunity, which under normal conditions is beneficial for the maintenance of good health, can become deteriorative under conditions of metabolic challenge, as exemplified by the immunosuppression characteristic of malnourished or starving individuals. It is now apparent that obesity is associated with a state of chronic inflammation, particularly in white adipose tissue. However, in the absence of obesity, infusion of animals with inflammatory cytokines or lipids can cause insulin resistance. It is possible that the stresses of obesity are similar enough to the stresses of an infection and the body reacts to obesity as it would to an infection. Atherosclerosis can be considered to have a significant chronic inflammatory component. Inflammation also contributes to the typical dyslipidemia associated with SLE that is characterized by elevations of VLDL, LDL and triglycerides as well as reduced HDL. The link between insulin resistance and SLE can be explained by the chronic inflammatory state, and the consequent dyslipoproteinemia. PMID- 17110318 TI - To smell the immune system: olfaction, autoimmunity and brain involvement. AB - Aside from its recognition and warning functions, olfaction serves many purposes in the CNS and remains one of the most important means of communication with the environment. In addition to olfactory tract input, the olfactory bulb also receives and provides input to other brain centers that modify neuronal activity. Research in the field of immunology as well as in various brain illnesses is beginning to indicate the increasing relevance of smell in pathophysiology. Much of this is based on the many intricate interactions that exist between the immune system and the nervous system, and evidence exists that there may be something unique about the olfactory system that is inextricably related to immunological function. In addition, accumulating evidence confirms the existence of olfactory dysfunction in brain disease, much of which appears at early stages including multiple sclerosis, Alzheimer's Disease, Parkinson's Disease, schizophrenia and depression. Such observations may further suggest that under certain circumstances, olfactory abnormalities may be associated with autoimmune conditions. Since the organization of the olfactory system is so sensitive, impairment may be noted at an early stage. This may become important in the prediction of certain brain illnesses. While preliminary evidence may suggest a role for olfaction in the management and alleviation of various disorders, investigation of its clinical relevance remains limited. PMID- 17110319 TI - Arrays of hope. AB - A large study by the MAQC consortium has established that different DNA microarray platforms can generate reproducible lists of differentially expressed genes. Now scientists are grappling with the challenges of moving the technology toward the clinic. PMID- 17110320 TI - American Idol and NIH grant review--redux. PMID- 17110321 TI - The anonymous American Idol manuscript reviewer. PMID- 17110322 TI - Grant review: American Idol or big brother? PMID- 17110324 TI - Knocking out the Argonautes. AB - Argonaute proteins are key players in gene silencing involving small RNAs. In this issue, Yigit et al. (2006) report a comprehensive study of Argonautes in the worm that places many of the 27 family members into a complex gene-silencing network. PMID- 17110325 TI - Neurodegeneration: a case of arrested development? AB - The neurodegenerative disease Spinocerebellar ataxia type 1 (SCA1) is a polyglutamine expansion disorder characterized by the death of Purkinje neurons in the brain. In this issue, Serra et al. (2006) implicate the impaired function of the orphan nuclear receptor RORalpha in SCA1 pathogenesis. Their intriguing results suggest that derailing a transcription program during embryonic development may render adult neurons more susceptible to toxic insults. PMID- 17110326 TI - Synaptic vesicles: an organelle comes of age. AB - Synaptic vesicles mediate the release of neurotransmitters at nerve terminals. In this issue of Cell, Takamori et al. (2006) analyze the lipid and protein components of synaptic vesicles, providing the most comprehensive description of synaptic vesicles to date. PMID- 17110327 TI - CFTR: new members join the fold. AB - The folding, misfolding, and degradation of membrane proteins is controlled by multiple processes within the cell. In this issue of Cell, Wang et al. (2006) present an interactome for CFTR, the chloride channel that is misfolded and prematurely degraded in cystic fibrosis. Among the proteins interacting with CFTR is a new member of the Hsp90 chaperone system, Aha1, that plays a central role in CFTR folding. PMID- 17110328 TI - A new function for p53 ubiquitination. AB - The amount of p53 protein in a cell is normally limited by ubiquitin-dependent degradation. In this issue of Cell, Le Cam et al. (2006) reveal that p53 ubiquitination contributes to transcriptional activation rather than protein stability. These results may provide insight into how p53 can modulate diverse cellular processes such as growth arrest and apoptosis. PMID- 17110329 TI - Cancer metastasis: building a framework. AB - Metastasis occurs when genetically unstable cancer cells adapt to a tissue microenvironment that is distant from the primary tumor. This process involves both the selection of traits that are advantageous to cancer cells and the concomitant recruitment of traits in the tumor stroma that accommodate invasion by metastatic cells. Recent conceptual and technological advances promote our understanding of the origins and nature of cancer metastasis. PMID- 17110330 TI - RORalpha-mediated Purkinje cell development determines disease severity in adult SCA1 mice. AB - Spinocerebellar ataxia type 1 (SCA1) is one of nine inherited, typically adult onset, polyglutamine neurodegenerative diseases. To examine whether development impacts SCA1, we used a conditional transgenic mouse model of SCA1 to delay the postnatal expression of mutant ATXN1 until after completion of cerebellar development. Delayed postnatal expression of mutant ATXN1 led to a substantial reduction in severity of disease in adults in comparison with early postnatal gene expression. This was linked to a destabilization of RORalpha, a transcription factor critical for cerebellar development. In SCA1 mice, there was a depletion of RORalpha and a reduction in expression of genes controlled by RORalpha. Partial loss of RORalpha enhanced mutant ATXN1 pathogenicity. Additionally, evidence points to the existence of a complex containing ATXN1, RORalpha, and the RORalpha coactivator Tip60. These studies indicate RORalpha and Tip60 have a role in SCA1 and suggest a mechanism by which compromising cerebellar development contributes to severity of neurodegeneration in an adult. PMID- 17110331 TI - The DNA damage machinery and homologous recombination pathway act consecutively to protect human telomeres. AB - Telomeres protect chromosome ends from being detected as lesions and from triggering DNA damage checkpoints. Paradoxically, telomere function depends on checkpoint proteins such as ATM and ATR, but a molecular model explaining this seemingly contradictory relationship has been missing so far. Here we show that the DNA damage machinery acts on telomeres in at least two independent steps. First, the ATR-dependent machinery is recruited to telomeres before telomere replication is completed, likely in response to single-stranded DNA resulting from replication fork stalling. Second, after replication, telomeres attract ATM and the homologous recombination (HR) machinery. In vivo and in vitro results suggest that the HR machinery is required for formation of a telomere-specific structure at chromosome ends after replication. Our results suggest that telomere ends need to be recognized as DNA damage to complete end replication and to acquire a structure that is essential for function. PMID- 17110332 TI - The highly conserved LepA is a ribosomal elongation factor that back-translocates the ribosome. AB - The ribosomal elongation cycle describes a series of reactions prolonging the nascent polypeptide chain by one amino acid and driven by two universal elongation factors termed EF-Tu and EF-G in bacteria. Here we demonstrate that the extremely conserved LepA protein, present in all bacteria and mitochondria, is a third elongation factor required for accurate and efficient protein synthesis. LepA has the unique function of back-translocating posttranslocational ribosomes, and the results suggest that it recognizes ribosomes after a defective translocation reaction and induces a back-translocation, thus giving EF-G a second chance to translocate the tRNAs correctly. We suggest renaming LepA as elongation factor 4 (EF4). PMID- 17110333 TI - Antisense transcription controls cell fate in Saccharomyces cerevisiae. AB - Entry into meiosis is a key developmental decision. We show here that meiotic entry in Saccharomyces cerevisiae is controlled by antisense-mediated regulation of IME4, a gene required for initiating meiosis. In MAT a/alpha diploids the antisense IME4 transcript is repressed by binding of the a1/alpha2 heterodimer at a conserved site located downstream of the IME4 coding sequence. MAT a/alpha diploids that produce IME4 antisense transcript have diminished sense transcription and fail to initiate meiosis. Haploids that produce the sense transcript have diminished antisense transcription and manifest several diploid phenotypes. Our data are consistent with transcription interference as a regulatory mechanism at the IME4 locus that determines cell fate. PMID- 17110334 TI - Analysis of the C. elegans Argonaute family reveals that distinct Argonautes act sequentially during RNAi. AB - Argonaute (AGO) proteins interact with small RNAs to mediate gene silencing. C. elegans contains 27 AGO genes, raising the question of what roles these genes play in RNAi and related gene-silencing pathways. Here we describe 31 deletion alleles representing all of the previously uncharacterized AGO genes. Analysis of single- and multiple-AGO mutant strains reveals functions in several pathways, including (1) chromosome segregation, (2) fertility, and (3) at least two separate steps in the RNAi pathway. We show that RDE-1 interacts with trigger derived sense and antisense RNAs to initiate RNAi, while several other AGO proteins interact with amplified siRNAs to mediate downstream silencing. Overexpression of downstream AGOs enhances silencing, suggesting that these proteins are limiting for RNAi. Interestingly, these AGO proteins lack key residues required for mRNA cleavage. Our findings support a two-step model for RNAi, in which functionally and structurally distinct AGOs act sequentially to direct gene silencing. PMID- 17110335 TI - Role for the PP2A/B56delta phosphatase in regulating 14-3-3 release from Cdc25 to control mitosis. AB - DNA-responsive checkpoints prevent cell-cycle progression following DNA damage or replication inhibition. The mitotic activator Cdc25 is suppressed by checkpoints through inhibitory phosphorylation at Ser287 (Xenopus numbering) and docking of 14-3-3. Ser287 phosphorylation is a major locus of G2/M checkpoint control, although several checkpoint-independent kinases can phosphorylate this site. We reported previously that mitotic entry requires 14-3-3 removal and Ser287 dephosphorylation. We show here that DNA-responsive checkpoints also activate PP2A/B56delta phosphatase complexes to dephosphorylate Cdc25 at a site distinct from Ser287 (T138), the phosphorylation of which is required for 14-3-3 release. However, phosphorylation of T138 is not sufficient for 14-3-3 release from Cdc25. Our data suggest that creation of a 14-3-3 "sink," consisting of phosphorylated 14-3-3 binding intermediate filament proteins, including keratins, coupled with reduced Cdc25-14-3-3 affinity, contribute to Cdc25 activation. These observations identify PP2A/B56delta as a central checkpoint effector and suggest a mechanism for controlling 14-3-3 interactions to promote mitosis. PMID- 17110336 TI - E4F1 is an atypical ubiquitin ligase that modulates p53 effector functions independently of degradation. AB - p53 is regulated by multiple posttranslational modifications, including Hdm2 mediated ubiquitylation that drives its proteasomal degradation. Here, we identify the p53-associated factor E4F1, a ubiquitously expressed zinc-finger protein first identified as a cellular target of the viral oncoprotein E1A, as an atypical ubiquitin E3 ligase for p53 that modulates its effector functions without promoting proteolysis. E4F1 stimulates oligo-ubiquitylation in the hinge region of p53 on lysine residues distinct from those targeted by Hdm2 and previously described to be acetylated by the acetyltransferase PCAF. E4F1 and PCAF mediate mutually exclusive posttranslational modifications of p53. E4F1 dependent Ub-p53 conjugates are associated with chromatin, and their stimulation coincides with the induction of a p53-dependent transcriptional program specifically involved in cell cycle arrest, and not apoptosis. Collectively, our data reveal that E4F1 is a key posttranslational regulator of p53, which modulates its effector functions involved in alternative cell fates: growth arrest or apoptosis. PMID- 17110337 TI - Crystal structure of the DsbB-DsbA complex reveals a mechanism of disulfide bond generation. AB - Oxidation of cysteine pairs to disulfide requires cellular factors present in the bacterial periplasmic space. DsbB is an E. coli membrane protein that oxidizes DsbA, a periplasmic dithiol oxidase. To gain insight into disulfide bond formation, we determined the crystal structure of the DsbB-DsbA complex at 3.7 A resolution. The structure of DsbB revealed four transmembrane helices and one short horizontal helix juxtaposed with Cys130 in the mobile periplasmic loop. Whereas DsbB in the resting state contains a Cys104-Cys130 disulfide, Cys104 in the binary complex is engaged in the intermolecular disulfide bond and captured by the hydrophobic groove of DsbA, resulting in separation from Cys130. This cysteine relocation prevents the backward resolution of the complex and allows Cys130 to approach and activate the disulfide-generating reaction center composed of Cys41, Cys44, Arg48, and ubiquinone. We propose that DsbB is converted by its specific substrate, DsbA, to a superoxidizing enzyme, capable of oxidizing this extremely oxidizing oxidase. PMID- 17110338 TI - Hsp90 cochaperone Aha1 downregulation rescues misfolding of CFTR in cystic fibrosis. AB - The pathways that distinguish transport of folded and misfolded cargo through the exocytic (secretory) pathway of eukaryotic cells remain unknown. Using proteomics to assess global cystic fibrosis (CF) transmembrane conductance regulator (CFTR) protein interactions (the CFTR interactome), we show that Hsp90 cochaperones modulate Hsp90-dependent stability of CFTR protein folding in the endoplasmic reticulum (ER). Cell-surface rescue of the most common disease variant that is restricted to the ER, DeltaF508, can be initiated by partial siRNA silencing of the Hsp90 cochaperone ATPase regulator Aha1. We propose that failure of DeltaF508 to achieve an energetically favorable fold in response to the steady-state dynamics of the chaperone folding environment (the "chaperome") is responsible for the pathophysiology of CF. The activity of cargo-associated chaperome components may be a common mechanism regulating folding for ER exit, providing a general framework for correction of misfolding disease. PMID- 17110339 TI - The architecture of the multisubunit TRAPP I complex suggests a model for vesicle tethering. AB - Transport protein particle (TRAPP) I is a multisubunit vesicle tethering factor composed of seven subunits involved in ER-to-Golgi trafficking. The functional mechanism of the complex and how the subunits interact to form a functional unit are unknown. Here, we have used a multidisciplinary approach that includes X-ray crystallography, electron microscopy, biochemistry, and yeast genetics to elucidate the architecture of TRAPP I. The complex is organized through lateral juxtaposition of the subunits into a flat and elongated particle. We have also localized the site of guanine nucleotide exchange activity to a highly conserved surface encompassing several subunits. We propose that TRAPP I attaches to Golgi membranes with its large flat surface containing many highly conserved residues and forms a platform for protein-protein interactions. This study provides the most comprehensive view of a multisubunit vesicle tethering complex to date, based on which a model for the function of this complex, involving Rab1-GTP and long, coiled-coil tethers, is presented. PMID- 17110340 TI - Molecular anatomy of a trafficking organelle. AB - Membrane traffic in eukaryotic cells involves transport of vesicles that bud from a donor compartment and fuse with an acceptor compartment. Common principles of budding and fusion have emerged, and many of the proteins involved in these events are now known. However, a detailed picture of an entire trafficking organelle is not yet available. Using synaptic vesicles as a model, we have now determined the protein and lipid composition; measured vesicle size, density, and mass; calculated the average protein and lipid mass per vesicle; and determined the copy number of more than a dozen major constituents. A model has been constructed that integrates all quantitative data and includes structural models of abundant proteins. Synaptic vesicles are dominated by proteins, possess a surprising diversity of trafficking proteins, and, with the exception of the V ATPase that is present in only one to two copies, contain numerous copies of proteins essential for membrane traffic and neurotransmitter uptake. PMID- 17110341 TI - The fly CAMTA transcription factor potentiates deactivation of rhodopsin, a G protein-coupled light receptor. AB - Control of membrane-receptor activity is required not only for the accuracy of sensory responses, but also to protect cells from excitotoxicity. Here we report the isolation of two noncomplementary fly mutants with slow termination of photoresponses. Genetic and electrophysiological analyses of the mutants revealed a defect in the deactivation of rhodopsin, a visual G protein-coupled receptor (GPCR). The mutant gene was identified as the calmodulin-binding transcription activator (dCAMTA). The known rhodopsin regulator Arr2 does not mediate this visual function of dCAMTA. A genome-wide screen identified five dCAMTA target genes. Of these, overexpression of the F box gene dFbxl4 rescued the mutant phenotypes. We further showed that dCAMTA is stimulated in vivo through interaction with the Ca(2+) sensor calmodulin. Our data suggest that calmodulin/CAMTA/Fbxl4 may mediate a long-term feedback regulation of the activity of Ca(2+)-stimulating GPCRs, which could prevent cell damage due to extra Ca(2+) influx. PMID- 17110342 TI - Positive association between STAT3 and Ki-67 in cervical intraepithelial neoplasia. AB - Signal transducer and activator of transcription 3 (STAT3) has been regarded as an oncogene in many types of cancers. However, its role in cervical carcinogenesis is not well determined. The purpose of this study was to evaluate the expression patterns of STAT3 in cervical intraepithelial neoplasia (CIN), normal cervix (NC), and squamous cell carcinoma (SCC) to explore its possible role in cervical carcinogenesis. Paraffin-embedded sections from 83 patients including 20 CIN 1, 10 CIN 2, 26 CIN 3, and 27 comparative cases of 10 NC and 17 stage Ib SCC were collected in this study. Immunohistochemistry was performed to analyze the expression patterns of STAT3, and the results obtained were categorized by a semiquantitative method and were further correlated with the CIN histopathologic grade and the proliferation marker, Ki-67, using the chi2 test. Our results showed that nuclear STAT3 expression was predominantly in the squamous epithelial cells, and that high-grade CIN and stage Ib SCC lesions had a higher nuclear STAT3 expression when compared with NC and CIN 1. Furthermore, the nuclear STAT3 expression in CIN was significantly correlated with Ki-67 expression (p = 0.025), but not CIN lesion grade. In summary, our results indicate that an altered STAT3 expression in CIN is correlated with cell proliferation but may not have a direct contribution to cervical carcinogenesis. PMID- 17110343 TI - Clinical characteristics of patients with sporadic colorectal cancer and primary cancers of other organs. AB - Most cancer patients often neglect the possibility of secondary cancer. Colorectal cancer (CRC) is the third leading cause of cancer death in Taiwan. It is important to be aware of the clinical characteristics of double cancer in CRC patients for early diagnosis and treatment. We retrospectively analyzed 1,031 CRC patients who underwent surgical treatment at the Department of Surgery of Kaohsiung Medical University Hospital between January 1998 and December 2004. Among these patients, CRC was accompanied by cancer of other organs in 17 patients (1.65%), either synchronously or metachronously. Therefore, we describe our experience regarding the location of CRC, the clinical symptoms and signs of these patients, the TNM stage, histology, phase, association with other malignancies, interval between cancers and clinical outcomes. Of the 17 patients in whom CRC was accompanied by primary cancer of other organs, there were four synchronous and 13 metachronous multiple cancer patients. Our patient group comprised six men and 11 women with ages ranging from 47 to 88 years (median age, 66 years). The most common location of CRC was the sigmoid colon. Six gastric cancers (35.2%) and six breast cancers (35.2%) were associated with primary CRC. The remaining six second primary cancers were one lung cancer, one thyroid cancer, one cervical cancer, one ovarian cancer, one skin cancer, and one urinary bladder cancer. Of the 13 metachronous multiple cancer patients, eight patients developed subsequent CRC after primary cancers of other organs, whereas two patients developed a subsequent second primary cancer after CRC. The intervals between the development of metachronous multiple cancers ranged from 2 to 19 years. In this retrospective analysis, breast and gastric cancer patients were at increased risk of developing subsequent secondary CRC. Careful attention should always be paid to the possibility of secondary CRC in treating these cancer patients. Cancer patients with hematochezia or gastrointestinal symptoms/signs should be evaluated for the possibility of second primary CRC during their regular follow-up. PMID- 17110344 TI - Morbidity after total thyroidectomy for benign thyroid disease: comparison of Graves' disease and non-Graves' disease. AB - The purpose of this study was to review the safety of total thyroidectomies for benign thyroid disease, with special emphasis on the comparison between Graves' disease and non-Graves' disease. In this study, 107 patients who underwent total thyroidectomies for clinically benign thyroid disease performed by the same surgeon between January 1987 and December 2004 were enrolled; 48 had Graves' disease and 59 had non-Graves' disease. The rates of temporary vs. permanent hypoparathyroidism, hematoma requiring surgical intervention, and temporary vs. permanent recurrent laryngeal nerve palsy (RLNP) after total thyroidectomy for benign thyroid disease were 34.6% vs. 3.7%, 6.5%, and 6.5% vs. 1.85%, respectively. The rates of permanent hypoparathyroidism and temporary RLNP in the Graves' disease group were significantly different when compared with the non Graves' disease group (8.3% vs. 0% and 11.5% vs. 2.5%, respectively). However, comparing the rates of temporary hypoparathyroidism, permanent RLNP, and postoperative hematoma, there was no statistically significant difference. Compared with total lobectomy, the rates of postoperative hematoma increased significantly for total thyroidectomy (6.5% vs. 0.48%). Total thyroidectomy for non-Graves' benign thyroid disease may be performed with minimal morbidity as has been advocated by many authors. For patients with Graves' disease in this study, however, the complication rates of permanent hypoparathyroidism and temporary RLNP were significantly increased. Therefore, we suggest that total thyroidectomy for Graves' disease should be performed by an experienced surgeon. PMID- 17110345 TI - Psychopathology of aboriginal and non-aboriginal adolescents living in the mountainous region of southern Taiwan. AB - The aim of this study was to examine the hypothesis that Taiwanese aboriginal adolescents feature more severe psychopathology than non-aboriginal adolescents who live in the same mountainous region of southern Taiwan, and to test the hypothesis by controlling other individual and environ-mental factors. In this study, a total of 251 aboriginal and 79 non-aboriginal Taiwanese adolescents were enrolled. Their psychopathology was measured by the Symptom Checklist-90-Revised Scale; demographic and family characteristics, and their affinity with their peer group and with their school were also assessed. The results of the multiple regression analysis revealed that aboriginal adolescents feature more severe psychopathology than non-aboriginal adolescents, and indicated that females and adolescents perceiving higher levels of family conflict and lower family support were more likely to experience more severe psychopathology than those perceiving the contrary. Those who devise strategies to improve the mental health of adolescents living in impoverished regions must take into consideration their ethnicity, gender, and family context when devising such treatment strategies. PMID- 17110346 TI - Safety of performing percutaneous dilational tracheostomy in patients with preexisting barotrauma. AB - Since its introduction in 1985 by Ciaglia et al, percutaneous dilational tracheostomy (PDT) has gradually become the procedure of choice in establishing a long-term airway in many intensive care units (ICU). However, the safety of performing PDT in patients with barotrauma is still unknown and has never been reported. We present the case of a 35-year-old man with AIDS, who was admitted to our medical ICU for pneumonia and acute respiratory distress syndrome. He developed subcutaneous emphysema and pneumomediastinum as complications of mechanical ventilation. After stabilization of the barotrauma, he underwent PDT with the standard Ciaglia Blue Rhino technique. However, rapid and extensive progression of preexisting barotraumas occurred shortly after PDT. This severe complication was nearly fatal. The prolonged procedure during which the susceptible lung was exposed to longer duration of high airway pressure was thought to be the mechanism of rapid deterioration of the preexisting barotrauma. With aggressive supportive care, the patient survived. To prevent further deterioration of preexisting barotraumas during and after PDT in future cases, we propose some principles that should be strictly followed. Under administration of these principles, we safely performed PDT in another case with preexisting barotrauma 1 month later. PMID- 17110347 TI - Adrenal leiomyoma treated by hand-assisted laparoscopic adrenalectomy: a case report. AB - A left adrenal incidentaloma was found by abdominal ultrasound and computed tomography (CT) in a 53-year-old woman who had a history of non-insulin dependent diabetes mellitus. Abdominal CT showed a well-circumscribed adrenal mass (5.5 x 4.5 x 3.5 cm) with low density (32HU) on precontrast scan and heterogeneous enhancement on postcontrast scan. Laboratory examinations revealed a nonfunctional adrenal tumor. As the malignant potential of the tumor could not be ruled out, the patient underwent hand-assisted laparoscopic adrenalectomy. Postoperative recovery was uneventful, and pathologic examination revealed leiomyoma. PMID- 17110348 TI - Freiberg's infraction --- treatment with metatarsal neck dorsal closing wedge osteotomy: report of two cases. AB - Freiberg's infraction is a disease of avascular necrosis that most commonly involves the head of the second or third metatarsal. Several mechanisms have been proposed for its pathogenesis, but stress overloading is the most widely accepted etiology. Nonoperative treatment is thought to be effective in the early stages but not in the late stages of the disease. The methods of operative treatment for symptomatic Freiberg's infraction remain controversial. We report two cases of late stage Freiberg's infraction treated by metatarsal neck dorsal closing wedge osteotomy with good results, and we infer that this operation may be recommended for patients with symptomatic Freiberg's infraction in whom conservative treatments have little effect. PMID- 17110349 TI - Conversion disorder in stroke: a case report. AB - Conversion disorder is caused by previous severe stress, emotional conflict, or an associated psychiatric disorder, and usually presents with one or more neurologic symptoms. Clinically, it is challenging to diagnose diseases such as transient ischemia attack, stroke, brain tumor, spinal cord injury, and neuropathy. In this case report, we present a male stroke patient who had a typical conversion disorder. PMID- 17110350 TI - Primary pulmonary synovial sarcoma: a case report. AB - We report a rare case of primary synovial sarcoma of the lung. A57-year-old man had a well-defined tumor in the right middle lobe seen on chest computed tomography, and underwent lobectomy. Grossly, the nonencapsulated tumor measured 4.5 cm in greatest diameter, with a solid and tan-white cut surface. Histologically, the tumor was mainly composed of a dense proliferation of spindle cells. Immunohistochemical studies were focally positive for epithelial membrane antigen, and diffusely positive for CD99 and Bcl-2. Cytokeratin, S-100 protein, desmin, smooth muscle act in, and CD34 were absent. SYT-SSX1 gene fusion transcript was detected by reverse transcription-polymerase chain reaction, which is diagnostic of primary synovial sarcoma of the lung. We also review the literature with regard to the clinicopathologic, immunohistochemical, and molecular studies of primary pulmonary synovial sarcoma. PMID- 17110351 TI - Impact of surface energy and roughness on cell distribution and viability. AB - The aim of this study was to assess the respective impacts of the surface energy and surface roughness of bare and coated steels on biofouling and sanitisation. Bioadhesion of Staphylococcus aureus CIP 53.154 was studied on two stainless steel surfaces with smooth or specific micro-topography. Two coatings were also studied: silicon oxide (hydrophilic) and polysiloxane (hydrophobic). On smooth surfaces, adhesion was reduced on an apolar coating and cell viability increased with the surface polarity. A specific micro-topography decreased the level of bacterial adhesion on bare surfaces by a factor ten. On this surface, only single adherent cells were observed, contrasting with cells in clusters on smoother surfaces. As a consequence, cell repartition influenced bacterial viability. Most isolated adherent cells were dead whereas cells in clusters were still alive. In addition, the quaternary ammonium chloride used in sanitisation, acted at once both as a tensio-active molecule and a biocide. It only displaced adherent cells but did not remove them. PMID- 17110352 TI - Improved estimates of environmental copper release rates from antifouling products. AB - The US Navy Dome method for measuring copper release rates from antifouling paint in-service on ships' hulls can be considered to be the most reliable indicator of environmental release rates. In this paper, the relationship between the apparent copper release rate and the environmental release rate is established for a number of antifouling coating types using data from a variety of available laboratory, field and calculation methods. Apart from a modified Dome method using panels, all laboratory, field and calculation methods significantly overestimate the environmental release rate of copper from antifouling coatings. The difference is greatest for self-polishing copolymer antifoulings (SPCs) and smallest for certain erodible/ablative antifoulings, where the ASTM/ISO standard and the CEPE calculation method are seen to typically overestimate environmental release rates by factors of about 10 and 4, respectively. Where ASTM/ISO or CEPE copper release rate data are used for environmental risk assessment or regulatory purposes, it is proposed that the release rate values should be divided by a correction factor to enable more reliable generic environmental risk assessments to be made. Using a conservative approach based on a realistic worst case and accounting for experimental uncertainty in the data that are currently available, proposed default correction factors for use with all paint types are 5.4 for the ASTM/ISO method and 2.9 for the CEPE calculation method. Further work is required to expand this data-set and refine the correction factors through correlation of laboratory measured and calculated copper release rates with the direct in situ environmental release rate for different antifouling paints under a range of environmental conditions. PMID- 17110353 TI - Biofouling communities on test panels coated with TBT and TBT-free copper based antifouling paints. AB - Formation of biofouling communities on the surfaces of steel panels coated with two different TBT-free copper-based and one TBT-based antifouling paint was examined following submersion for periods of 3, 6, 9 and 12 months in Kastela Bay, Croatia. Test panels coated with Shopprimer and anticorrosive paint were used as control panels. Thirty five taxa of benthic algae and 32 taxa of benthic animals were found. Diatomeae dominated by frequency and abundance on test panels coated with antifouling paints, while the animal species Serpula vermicularis, Mytilus galloprovincialis and Balanus amphitrite amphitrite dominated the fouling communities on control panels. On panels protected by antifouling paints, low values of biomass were found compared to the very high values associated with control panels. PMID- 17110354 TI - A methodology for evaluating biocide release rate, surface roughness and leach layer formation in a TBT-free, self-polishing antifouling coating. AB - Due to the forthcoming IMO ban on the use of tributyltin (TBT) antifouling paints, a new generation of TBT-free coatings has been developed that typically contain cuprous oxide and an organic co-biocide. Accurate and reproducible test methods are needed to evaluate the performance and environmental impact of these new coatings. This study investigated a methodology for evaluating TBT-free, AF coatings containing cuprous oxide. A commercially available AF coating underwent rotary immersion testing at 0, 0.51 and 2.05 m s-1. Scanning electron microscopy (SEM) and energy dispersive x-ray (EDX) analysis were used to assess leach layer formation, percentage cuprous oxide by weight and particle size distribution (PSD). Biocide release rates and surface roughness were also measured. An increase in rotary speed caused a spike in Cu2+ release rate after which the release rate stabilised to previous levels. An increase in leach layer thickness was also observed after the rotary speed increase. A model is suggested to account for the observations. PMID- 17110355 TI - Antifouling potential of cyanobacteria: a mini-review. AB - Cyanobacteria produce a variety of bioactive metabolites that may have allelochemical functions in the natural environment, such as in the prevention of fouling by colonising organisms. Chemical compounds from cyanobacteria are also of biotechnological interest, especially for clinical applications, because of their antibiotic, algicidal, cytotoxic, immunosupressive and enzyme inhibiting activities. Cyanobacterial metabolites have the potential for use in antifouling technology, since they show antibacterial, antialgal, antifungal and antimacrofouling properties which could be expoited in the prevention of biofouling on man-made substrata in the aquatic environment. Molecules with antifouling activity represent a number of types including fatty acids, lipopeptides, amides, alkaloids, terpenoids, lactones, pyrroles and steroids. The isolation of biogenic compounds and the determination of their structure may provide leads for future development of, for example, environmentally friendly antifouling paints. An advantage of exploring the efficacy of cyanobacterial products is that the organisms can be grown in mass culture, which can be manipulated to achieve optimal production of bioactive substances. Phycotoxins and related products from cyanobacteria may serve as materials for antimicro- and antimacrofouling applications. A survey of antibiotic compounds with antifouling potential revealed more than 21 different antifouling substances from 27 strains of cyanobacteria. PMID- 17110356 TI - Adhesive mechanisms in cephalopods: a review. AB - Several genera of cephalopods (Nautilus, Sepia, Euprymna and Idiosepius) produce adhesive secretions, which are used for attachment to the substratum, for mating and to capture prey. These adhesive structures are located in different parts of the body, viz. in the digital tentacles (Nautilus), in the ventral surface of the mantle and fourth arm pair (Sepia), in the dorsal epidermis (Euprymna), or in the dorsal mantle side and partly on the fins (Idiosepius). Adhesion in Sepia is induced by suction of dermal structures on the mantle, while for Nautilus, Euprymna and Idiosepius adhesion is probably achieved by chemical substances. Histochemical studies indicate that in Nautilus and Idiosepius secretory cells that appear to be involved in adhesion stain for carbohydrates and protein, whilst in Euprymna only carbohydrates are detectable. De-adhesion is either achieved by muscle contraction of the tentacles and mantle (Nautilus and Sepia) or by secretion of substances (Euprymna). The de-adhesive mechanism used by Idiosepius remains unknown. PMID- 17110357 TI - Recent developments in superhydrophobic surfaces and their relevance to marine fouling: a review. AB - In this review, a brief synopsis of superhydrophobicity (i.e. extreme non wettability) and its implications on marine fouling are presented. A short overview of wettability and recent experimental developments aimed at fabricating superhydrophobic surfaces by tailoring their chemical nature and physical appearance (i.e. substratum texture) are reviewed. The formation of responsive/"smart" surfaces, which adjust their physico-chemical properties to variations in some outside physical stimulus, including light, temperature, electric field, or solvent, is also described. Finally, implications of tailoring the surface chemistry, texture, and responsiveness of surfaces on the design of effective marine fouling coatings are considered and discussed. PMID- 17110358 TI - Molecular mechanisms of transcriptional regulation by nuclear receptors. Perspectives for therapeutic implications. AB - Nuclear receptors are ligand-regulated transcription factors that evolved from an ancestral orphan receptor into a highly diverse family present throughout the entire animal kingdom. They encompass receptors for steroid and non-steroid hormones, vitamins and metabolic intermediates. These receptors signal through endocrine, paracrine, autocrine and intracrine networks to regulate multiple aspects of animal physiology, including homeostasis, development and reproduction. They exert genomic effects via direct binding as monomers, homo- or heterodimers on cognate DNA elements (hormone response elements). They also participate in signal transduction cross-talk to indirectly modulate other gene expression programmes. By coordinating expression of genetic programmes, nuclear receptors contribute to cell fate-determining processes, thereby shaping and sustaining the organism. All these actions result from one fundamental interaction: receptor binding of a cognate ligand, which induces a major allosteric change in the ligand-binding domain. This conformational alteration is transformed into cascades of protein-protein recognitions, culminating in the establishment of coregulator/cointegrator complexes on gene promoters. Coregulators induce chromatin remodelling and acetylation, thus enabling the targeted recruitment and activation of the basal transcription machinery. This review discusses the molecular infrastructure of nuclear receptor signalling. Emphasis is given to determinants of signalling specificity, especially since they highlight prominent targets for novel drug discovery. PMID- 17110359 TI - Novel approaches to the management of graves' ophthalmopathy. AB - Severe Graves' ophthalmopathy constitutes a complex therapeutic challenge and treatment outcome often is not satisfactory. Established methods of treatment include high-dose glucocorticoids, orbital radiotherapy and orbital decompression. Recently, the use of intravenous glucocorticoids has been shown to provide more favorable results than oral glucocorticoids. Novel treatments under investigation include somatostatin analogues, intravenous immunoglobulins and antioxidants. Low-dose immunosuppressive drugs (namely cyclosporine and, possibly, methotrexate) might be useful as an adjunct to established methods, particularly in view of a glucocorticoid-sparing action. Because cytokines play an important role in the pathogenesis of the disease, cytokine antagonists, which are currently evaluated in rheumatoid arthritis and other autoimmune conditions, might constitute in the future a valuable tool for the management of eye disease. Prevention of Graves' ophthalmopathy would be desirable, but so far it is limited to secondary prevention (arrest of progression of subclinical disease to clinical disease) and tertiary prevention (avoidance of deterioration or complications of clinical disease): among preventive measures smoking withdrawal is probably the most important one. Primary prevention (in the absence of disease) is only speculative, but oral tolerance induction or vaccination with the offending antigen(s) might prove beneficial for prevention of Graves' ophthalmopathy in genetically susceptible individuals. PMID- 17110360 TI - Influence of cigarette smoking on thyroid function, goiter formation and autoimmune thyroid disorders. AB - Both smoking and thyroid dysfunction are frequent in the general population. Many studies have shown that cigarette smoking interferes with thyroid function and with the evolution of thyroid pathology (e.g. goiter formation and thyroid cancer development). Some studies have also suggested a significant correlation of Graves' hyperthyroidism and Graves' ophthalmopathy with the severity of smoking. In addition, cigarette smoking may reduce the effectiveness of some therapeutic modalities, such as orbital radiotherapy or high-dose systemic glucocorticoid administration for severe thyroid eye disease. Tobacco smoking seems to induce similar changes in thyroid function in the adult and the fetus. This review article discusses the effect of cigarette smoking on thyroid function and morphology as well as on thyroid autoimmunity. PMID- 17110361 TI - Long term thyroid function after (131)I treatment for toxic adenoma. AB - Radioactive iodine is a widely used treatment for hyperthyroidism caused by solitary autonomously functioning thyroid nodule (toxic adenoma). The aim of this retrospective analysis is to report the long term effects of this therapy on the thyroid function of patients with toxic adenoma treated in our department. Between 1968 and 1996, 160 patients received a single dose of (131)I (range 25-40 mCi) for hyperthyroidism caused by toxic adenoma. In 126 of these (110 females, 26 males), follow-up was feasible either in our Endocrine Outpatient Clinic or through correspondence. The mean observation period was 5.3 years (range 1-21 years, median 4.0). Post treatment evaluation revealed that: a) 57 patients became euthyroid and remained free of disease up to the last visit (mean observation period 5.76+/-0.52 years, range 1-21 years, median 5 years), b) 69 patients developed hypothyroidism, all within 1 to 12 months (5.9+/-0.49 months), c) persistence or recurrence of the disease (ie. thyrotoxicosis) was not observed, d) the (131)I dose, or the (131)I pretreatment TSH levels were not different between patients who developed hypothyroidism and those who became and remained euthyroid. CONCLUSION: 131I administration in the above-mentioned dose to patients with toxic adenoma: a) was a safe and very effective therapy, and b) led to hypothyroidism which developed within the first year after (131)I administration in 55% of the patients. PMID- 17110362 TI - Normalization of thyroid hormone levels in patients with either hyper- or hypothyroidism results in a profound change of atrial natriuretic peptide (ANP) levels. AB - Atrial Natriuretic Peptide (ANP) is a hormone produced by cardiac atrial myocytes. Thyroid hormones may affect its release. The aim of this study was to analyze the effect of hyper and hypothyroidism on the secretion of ANP and its relationship with the changes of the renin-angiotensin-aldosterone system. Plasma ANP concentration as well as plasma renin activity (PRA) and aldosterone (ALDO) were measured in 21 patients with hyperthyroidism, 12 patients with hypothyroidism and 29 normal controls. Moreover, in 5 hyperthyroid and 8 hypothyroid patients hormones measurements were performed at diagnosis and 15, 30, 45 and 60 days after starting the appropriate therapy. Statistical analysis was performed employing the Wilcoxon two sample test to compare the ANP levels in normal and hyper or hypothyroid patients while the pairwise comparisons were evaluated using the Spermans rank correlation. Partial correlation and regression models were used to examine the joint effects of multiple predictors on ANP. ANP levels were significantly higher in hyperthyroid (mean+/-SEM, 17+/-1.7 pmol/L, +/ <0.001) and lower in hypothyroid patients (6.18+/-1.14 pmol/L, p<0.01) compared to normals (10.35+/-0.59 pmol/L). In the hyperthyroid patients following treatment with antithyroid drugs plasma ANP levels reached normal values after 15 days and remained stable thereafter. In the hypothyroid patients, ANP levels increased gradually following initiation of treatment with thyroxine, reaching values comparable to controls at day 45. PRA was significantly higher only in hyperthyroid patients compared to controls. ALDO concentrations were not different in all groups studied. No correlation was found between plasma ANP levels and T3, T4, TSH, PRA and ALDO levels in the individual groups, while a positive correlation for T3 (r=0.610, p<0.01), T4 (r=0.653, p<0.01) and pulse rate (r=0.704, p<0.01) and negative correlation with TSH (r=-0.519, p<0.01) was found when all groups were pooled together. In conclusion, our data indicate that thyroid hormones affect positively ANP secretion from cardiac myocytes. This effect may be, at least in part, indirect via hemodynamic alterations. PMID- 17110363 TI - Multiple pituitary hormone abnormalities, fever, behavioral problems, seizures and apnoic spells in a 6-year old girl. AB - A 6-year old girl was examined having two years previously presented a transient Cushing's syndrome, followed by recurrent hyponatremia, attributed to inappropriate ADH secretion (SIADH). The brain MRI showed no abnormalities on repeated examinations, except for a suggestion of empty sella syndrome. During the past two years she also presented recurrent episodes of a prolonged febrile illness of unknown origin. All investigations related to infectious, autoimmune neoplastic diseases, histiocytosis-X or neurosarcoidosis were negative and the fever was characterized as central. The patient also presented episodes of tonic clonic seizures, myoclonias and behavioral problems (alternating states of irritability, sleepiness and apathy, optic and hearing illusions and phobias) with or without hyponatremia. Her cerebrospinal fluid (CSF) examination was not indicative of encephalitis and the encephalogram (EEG) showed only slowing of background activity. At the age of 4.75 years she, started to have recurrent episodes of hypopnoea/apnoea with severe desaturation and hypercapnia, occasionally requiring intubation and ventilation. She also developed unilateral miosis corneal ulceration and bilateral ptosis (oculo-sympathetic paresis). Repeat brain MRI and CT scans of the mediastinum excluded organic causes of apnoeas and of oculo-sympathetic paresis, such as neuroblastoma or lymphoma. Furthermore, on a 24 hour electrocardiogram recording, using power spectral analysis, significantly reduced heart rate variability was observed, by comparison with age-specific normal ranges. Thus the apnoeas, ptosis, miosis and temperature instability were attributed to autonomic dysfunction. During the same period, the patient presented significant growth retardation and growth hormone (GH) deficiency was confirmed during two provocative tests (peak GH levels: 3.1 and 2.9 ng/ml (normal>10). Moreover, thyrotropin (TSH) deficiency and persistent hyperprolactinemia were detected. She was started on hGH and thyroxine. She was also put on fluid restriction and increased sodium intake for her SIADH. She was advised to use O2 administration by mask in case of apnoeas. The child died at age 6 6/12 years as a result of apnoea during sleep. IN CONCLUSION: Multiple pituitary hormonal abnormalities, together with symptoms of autonomic neuropathy (apnoeas, ptosis, miosis, tachycardia, temperature instability) and encephalopathy (seizures, myoclonias and behavioral problems) developed in a 4 year old girl. The suggested diagnoses were: 1. Neurometabolic disorder, 2. Mitochondrial disorder, 3. Post infectious autoimmune process. PMID- 17110364 TI - Homeostasis. The ancient Greek origin of a modern scientific principle. PMID- 17110365 TI - Striped Smith-Waterman speeds database searches six times over other SIMD implementations. AB - MOTIVATION: The only algorithm guaranteed to find the optimal local alignment is the Smith-Waterman. It is also one of the slowest due to the number of computations required for the search. To speed up the algorithm, Single Instruction Multiple-Data (SIMD) instructions have been used to parallelize the algorithm at the instruction level. RESULTS: A faster implementation of the Smith Waterman algorithm is presented. This algorithm achieved 2-8 times performance improvement over other SIMD based Smith-Waterman implementations. On a 2.0 GHz Xeon Core 2 Duo processor, speeds of >3.0 billion cell updates/s were achieved. AVAILABILITY: http://farrar.michael.googlepages.com/Smith-waterman PMID- 17110366 TI - Network neighborhood analysis with the multi-node topological overlap measure. AB - MOTIVATION: The goal of neighborhood analysis is to find a set of genes (the neighborhood) that is similar to an initial 'seed' set of genes. Neighborhood analysis methods for network data are important in systems biology. If individual network connections are susceptible to noise, it can be advantageous to define neighborhoods on the basis of a robust interconnectedness measure, e.g. the topological overlap measure. Since the use of multiple nodes in the seed set may lead to more informative neighborhoods, it can be advantageous to define multi node similarity measures. RESULTS: The pairwise topological overlap measure is generalized to multiple network nodes and subsequently used in a recursive neighborhood construction method. A local permutation scheme is used to determine the neighborhood size. Using four network applications and a simulated example, we provide empirical evidence that the resulting neighborhoods are biologically meaningful, e.g. we use neighborhood analysis to identify brain cancer related genes. AVAILABILITY: An executable Windows program and tutorial for multi-node topological overlap measure (MTOM) based analysis can be downloaded from the webpage (http://www.genetics.ucla.edu/labs/horvath/MTOM/). PMID- 17110367 TI - GARD: a genetic algorithm for recombination detection. AB - MOTIVATION: Phylogenetic and evolutionary inference can be severely misled if recombination is not accounted for, hence screening for it should be an essential component of nearly every comparative study. The evolution of recombinant sequences can not be properly explained by a single phylogenetic tree, but several phylogenies may be used to correctly model the evolution of non recombinant fragments. RESULTS: We developed a likelihood-based model selection procedure that uses a genetic algorithm to search multiple sequence alignments for evidence of recombination breakpoints and identify putative recombinant sequences. GARD is an extensible and intuitive method that can be run efficiently in parallel. Extensive simulation studies show that the method nearly always outperforms other available tools, both in terms of power and accuracy and that the use of GARD to screen sequences for recombination ensures good statistical properties for methods aimed at detecting positive selection. AVAILABILITY: Freely available http://www.datamonkey.org/GARD/ PMID- 17110368 TI - SAGA: a subgraph matching tool for biological graphs. AB - MOTIVATION: With the rapid increase in the availability of biological graph datasets, there is a growing need for effective and efficient graph querying methods. Due to the noisy and incomplete characteristics of these datasets, exact graph matching methods have limited use and approximate graph matching methods are required. Unfortunately, existing graph matching methods are too restrictive as they only allow exact or near exact graph matching. This paper presents a novel approximate graph matching technique called SAGA. This technique employs a flexible model for computing graph similarity, which allows for node gaps, node mismatches and graph structural differences. SAGA employs an indexing technique that allows it to efficiently evaluate queries even against large graph datasets. RESULTS: SAGA has been used to query biological pathways and literature datasets, which has revealed interesting similarities between distinct pathways that cannot be found by existing methods. These matches associate seemingly unrelated biological processes, connect studies in different sub-areas of biomedical research and thus pose hypotheses for new discoveries. SAGA is also orders of magnitude faster than existing methods. AVAILABILITY: SAGA can be accessed freely via the web at http://www.eecs.umich.edu/saga. Binaries are also freely available at this website. PMID- 17110369 TI - Tree and rate estimation by local evaluation of heterochronous nucleotide data. AB - MOTIVATION: Heterochronous gene sequence data is important for characterizing the evolutionary processes of fast-evolving organisms such as RNA viruses. A limited set of algorithms exists for estimating the rate of nucleotide substitution and inferring phylogenetic trees from such data. The authors here present a new method, Tree and Rate Estimation by Local Evaluation (TREBLE) that robustly calculates the rate of nucleotide substitution and phylogeny with several orders of magnitude improvement in computational time. METHODS: For the basis of its rate estimation TREBLE novelly utilizes a geometric interpretation of the molecular clock assumption to deduce a local estimate of the rate of nucleotide substitution for triplets of dated sequences. Averaging the triplet estimates via a variance weighting yields a global estimate of the rate. From this value, an iterative refinement procedure relying on statistical properties of the triplets then generates a final estimate of the global rate of nucleotide substitution. The estimated global rate is then utilized to find the tree from the pairwise distance matrix via an UPGMA-like algorithm. RESULTS: Simulation studies show that TREBLE estimates the rate of nucleotide substitution with point estimates comparable with the best of available methods. Confidence intervals are comparable with that of BEAST. TREBLE's phylogenetic reconstruction is significantly improved over the other distance matrix method but not as accurate as the Bayesian algorithm. Compared with three other algorithms, TREBLE reduces computational time by a minimum factor of 3000. Relative to the algorithm with the most accurate estimates for the rate of nucleotide substitution (i.e. BEAST), TREBLE is over 10,000 times more computationally efficient. AVAILABILITY: jdobrien.bol.ucla.edu/TREBLE.html PMID- 17110370 TI - Indelign: a probabilistic framework for annotation of insertions and deletions in a multiple alignment. AB - MOTIVATION: A quantitative study of molecular evolutionary events such as substitutions, insertions and deletions from closely related genomes requires (1) an accurate multiple sequence alignment program and (2) a method to annotate the insertions and deletions that explain the 'gaps' in the alignment. Although the former requirement has been extensively addressed, the latter problem has received little attention, especially in a comprehensive probabilistic framework. RESULTS: Here, we present Indelign, a program that uses a probabilistic evolutionary model to compute the most likely scenario of insertions and deletions consistent with an input multiple alignment. It is also capable of modifying the given alignment so as to obtain a better agreement with the evolutionary model. We find close to optimal performance and substantial improvement over alternative methods, in tests of Indelign on synthetic data. We use Indelign to analyze regulatory sequences in Drosophila, and find an excess of insertions over deletions, which is different from what has been reported for neutral sequences. AVAILABILITY: The Indelign program may be downloaded from the website http://veda.cs.uiuc.edu/indelign/ SUPPLEMENTARY INFORMATION: Supplementary material is available at Bioinformatics online. PMID- 17110371 TI - In vitro efficacy, resistance selection, and structural modeling studies implicate the malarial parasite apicoplast as the target of azithromycin. AB - Azithromycin (AZ), a broad-spectrum antibacterial macrolide that inhibits protein synthesis, also manifests reasonable efficacy as an antimalarial. Its mode of action against malarial parasites, however, has remained undefined. Our in vitro investigations with the human malarial parasite Plasmodium falciparum document a remarkable increase in AZ potency when exposure is prolonged from one to two generations of intraerythrocytic growth, with AZ producing 50% inhibition of parasite growth at concentrations in the mid to low nanomolar range. In our culture-adapted lines, AZ displayed no synergy with chloroquine (CQ), amodiaquine, or artesunate. AZ activity was also unaffected by mutations in the pfcrt (P. falciparum chloroquine resistance transporter) or pfmdr1 (P. falciparum multidrug resistance-1) drug resistance loci, as determined using transgenic lines. We have selected mutant, AZ-resistant 7G8 and Dd2 parasite lines. In the AZ-resistant 7G8 line, the bacterial-like apicoplast large subunit ribosomal RNA harbored a U438C mutation in domain I. Both AZ-resistant lines revealed a G76V mutation in a conserved region of the apicoplast-encoded P. falciparum ribosomal protein L4 (PfRpl4). This protein is predicted to associate with the nuclear genome-encoded P. falciparum ribosomal protein L22 (PfRpl22) and the large subunit rRNA to form the 50 S ribosome polypeptide exit tunnel that can be occupied by AZ. The PfRpl22 sequence remained unchanged. Molecular modeling of mutant PfRpl4 with AZ suggests an altered orientation of the L75 side chain that could preclude AZ binding. These data imply that AZ acts on the apicoplast bacterial-like translation machinery and identify Pfrpl4 as a potential marker of resistance. PMID- 17110372 TI - Membrane cholesterol content modulates ClC-2 gating and sensitivity to oxidative stress. AB - ClC-2 is a broadly expressed member of the voltage-gated ClC chloride channel family. In this study, we aimed to evaluate the role of the membrane lipid environment in ClC-2 function, and in particular the effect of cholesterol and ClC-2 distribution in membrane microdomains. Detergent-resistant and detergent soluble microdomains (DSM) were isolated from stably transfected HEK293 cells by a discontinuous OptiPrep gradient. ClC-2 was found concentrated in detergent insoluble membranes in basal conditions and relocalized to DSM upon cholesterol depletion by methyl-beta-cyclodextrin. As assessed by patch clamp recordings, relocalization was accompanied by acceleration of the activation kinetics of the channel. A similar distribution and activation pattern were obtained when cells were treated with the oxidant tert-butyl hydroperoxide and after ATP depletion. In both cases activation was prevented by cholesterol enrichment of cells. We conclude that the cholesterol environment regulates ClC-2 activity, and we provide evidence that the increase in ClC-2 activity in response to acute oxidative or metabolic stress involves relocalization of this channel to DSM. PMID- 17110373 TI - Evidence for two modes of development of acquired tumor necrosis factor-related apoptosis-inducing ligand resistance. Involvement of Bcl-xL. AB - Previous studies have shown that repeated application of TRAIL induces acquired resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Using human prostate adenocarcinoma DU-145 and human pancreatic carcinoma MiaPaCa 2 cells as a model, we now demonstrate for the first time that two states of acquired TRAIL resistance can be developed after TRAIL treatment. Data from survival assay and Western blot analysis show that acquired TRAIL resistance was developed within 1 day and gradually decayed within 6 days after TRAIL treatment in both cell lines. After TRAIL treatment, the level of Bcl-xL increased and reached a maximum within 2 days and gradually decreased in both cell lines. Bcl xL-mediated development of acquired TRAIL resistance was suppressed by knockdown of Bcl-xL expression. Protein interaction assay revealed that during the development of TRAIL resistance, Bcl-xL dissociated from Bad and then associated with Bax. Overexpression of mutant-type Bad (S136A), which prevents this dissociation, partially suppressed the development of acquired TRAIL resistance. Thus, our results suggest that (a) dissociation of Bad from Bcl-xL and (b) an increase in the intracellular level of Bcl-xL are responsible for development of acquired TRAIL resistance. PMID- 17110374 TI - Insertion and topology of normal and mutant bestrophin-1 in the endoplasmic reticulum membrane. AB - The vitelliform macular dystrophy type 2 (VMD2) gene mutated in Best macular dystrophy encodes a 585-amino acid putative transmembrane protein termed bestrophin-1. The vast majority of known disease-associated alterations are of the missense type, which cluster near predicted transmembrane domains (TMDs). To investigate bestrophin-1 membrane topology and to assess consequences of point mutations on membrane integration, we have analyzed the insertion of putative TMDs into the endoplasmic reticulum (ER) membrane. Out of six potential TMDs, our data suggest a topological model of bestrophin-1 with four transmembrane-spanning segments and one large cytoplasmatic loop between putative TMD2 and TMD5. Consequently, a relatively hydrophobic segment containing putative TMD3 (aa 130 149) and TMD4 (aa 179-201) is located within the cytoplasm. Furthermore, we show that three out of 18 disease-associated alterations investigated (I73N, Y85H, F281del) reveal measurable effects on membrane insertion suggesting that defective membrane integration of bestrophin-1 may represent a potential disease mechanism for a small subset of Best macular dystrophy-related mutations. PMID- 17110375 TI - Phosphoinositide 3-kinase C2alpha links clathrin to microtubule-dependent movement. AB - Phosphoinositide 3-kinase C2alpha (PI3K-C2alpha) is a type II PI-3-kinase that has been implicated in several important membrane transport and signaling processes. We previously found that overexpression of PI3K-C2alpha inhibits clathrin-mediated membrane trafficking and induces proliferation of novel clathrin-coated structures within the cytoplasm. Using fluorescently tagged fusions of PI3K-C2alpha and clathrin, we explored the behavior of these structures in intact cells. Both proteins are present in the structures, and using rapid image acquisition and fluorescence photoactivation probes, we find that they exhibit localized, rapid mobility (5-20 microm/s). The movement is micro-tubule-based as revealed by use of inhibitors, and PI3K-C2alpha accumulates on microtubules rapidly and reversibly following cytoplasmic acidification, which also blocks movement. Dynactin mediates the movement of these clathrin-PI3K C2alpha structures, since disruption of dynactin function by overexpression of its p50 subunit also inhibits movement. Finally, immunoprecipitation experiments reveal an interaction between endogenous PI3K-C2alpha and dynactin subunits. Together, these results reveal a molecular linkage between PI3K-C2alpha and the microtubule motor machinery, with implications for membrane trafficking in intact cells. PMID- 17110376 TI - CCAAT/enhancer-binding protein (C/EBP) beta is acetylated at multiple lysines: acetylation of C/EBPbeta at lysine 39 modulates its ability to activate transcription. AB - Transcription factor function can be modulated by post-translational modifications. Because the transcription factor CCAAT/enhancer-binding protein (C/EBP) beta associates with the nuclear coactivator p300, which contains acetyltransferase activity, acetylation of C/EBPbeta was examined to understand its regulation and function. C/EBPbeta is acetylated by acetyltransferases p300 and p300/CREB-binding protein associated factor. Endogenous C/EBPbeta in 3T3 F442A preadipocytes is also recognized by an acetyl-lysine-specific antibody. Analysis of truncations of C/EBPbeta and peptides based on C/EBPbeta sequences identified multiple lysines within C/EBPbeta that can be acetylated. Among these, a novel acetylation site at lysine 39 of C/EBPbeta was identified. Mutation of Lys-39 to arginine or alanine impairs its acetylation and the ability of C/EBPbeta to activate transcription at the promoters for C/EBPalpha and c-fos. Different C/EBPbeta-responsive promoters require different patterns of acetylated lysines in C/EBPbeta for transcription activation. Furthermore, C/EBPbeta acetylation was increased by growth hormone, and mutation of Lys-39 impaired growth hormone-stimulated c-fos promoter activation. These data suggest that acetylation of Lys-39 of C/EBPbeta, alone or in combination with acetylation at other lysines, may play a role in C/EBPbeta-mediated transcriptional activation. PMID- 17110377 TI - Distinct patterns of cytokine regulation of APOBEC3G expression and activity in primary lymphocytes, macrophages, and dendritic cells. AB - Human APOBEC3G (A3G), a deoxycytidine deaminase, is a broadly acting antiretroviral factor expressed in a variety of cells. Mitogen activation of CD4 T cells enhances A3G expression and leads to recruitment of low molecular mass (LMM) A3G, which functions as a post-entry human immunodeficiency virus (HIV) restriction factor, into enzymatically inactive, high molecular mass (HMM) RNA protein complexes that include Staufen RNA-transporting granules. We now report that interleukin-2 (IL-2), IL-15 and, to a lesser extent, IL-7 enhance the expression of A3G in peripheral blood lymphocytes and that this effect is blocked by inhibitors of the JAK and MAPK signaling pathways. In mixed cultures of CD4+ T cells containing either HMM or LMM A3G, HIV preferentially infected cells containing HMM A3G. A3G shifted into a HMM complex when IL-2, -7, or -15 was added to resting T cells, likely explaining how cytokine treatment renders resting CD4+ T cells permissive to HIV infection. Similarly, poly(I:C)/tumor necrosis factor-alpha-induced maturation of dendritic cells was associated with a sharp increase in A3G expression; however, this induction led to the accumulation of LMM A3G. Together, these results highlight the distinct inductive effects of select cytokines on A3G gene expression and A3G complex assembly that occur in natural cellular targets of HIV infection. PMID- 17110378 TI - Targeted cyclooxygenase gene (ptgs) exchange reveals discriminant isoform functionality. AB - The prostaglandin G/H synthase enzymes, commonly termed COX-1 and COX-2, differ markedly in their responses to regulatory stimuli and their tissue expression patterns. COX-1 is the dominant source of "housekeeping" prostaglandins, whereas COX-2 synthesizes prostaglandins of relevance to pain, inflammation, and mitogenesis. Despite these distinctions, the two enzymes are remarkably conserved, and their subcellular distributions overlap considerably. To address the functional interchangeability of the two isozymes, mice in which COX-1 is expressed under COX-2 regulatory elements were created by a gene targeting "knock in" strategy. In macrophages from these mice, COX-1 was shown to be lipopolysaccharide-inducible in a manner analogous to COX-2 in wild-type macrophages. However, COX-1 failed to substitute effectively for COX-2 in lipopolysaccharide-induced prostaglandin E2 synthesis at low concentrations of substrate and in the metabolism of the endocannabinoid 2-arachidonylglycerol. The marked depression of the major urinary metabolite of prostacyclin in COX-2 null mice was only partially rescued by COX-1 knock-in, whereas the main urinary metabolite of prostaglandin E2 was rescued totally. Replacement with COX-1 partially rescued the impact of COX-2 deletion on reproductive function. The renal pathology consequent to COX-2 deletion was delayed but not prevented, whereas the corresponding peritonitis was unaltered. Insertion of COX-1 under the regulatory sequences that drive COX-2 expression indicated that COX-1 can substitute for some COX-2 actions and rescue only some of the consequences of gene disruption. Manipulation of COX-2 also revealed a preference for coupling with distinct downstream prostaglandin synthases in vivo. These mice will provide a valuable reagent with which to elucidate the distinct roles of the COX enzymes in mammalian biology. PMID- 17110379 TI - A novel nuclear interactor of ARF and MDM2 (NIAM) that maintains chromosomal stability. AB - The ARF tumor suppressor signals through p53 and other poorly defined anti proliferative pathways to block carcinogenesis. In a search for new regulators of ARF signaling, we discovered a novel nuclear protein that we named NIAM (nuclear interactor of ARF and MDM2) for its ability to bind both ARF and the p53 antagonist MDM2. NIAM protein is normally expressed at low to undetectable levels in cells because of, at least in part, MDM2-mediated ubiquitination and proteasomal degradation. When reintroduced into cells, NIAM activated p53, caused a G1 phase cell cycle arrest, and collaborated with ARF in an additive fashion to suppress proliferation. Notably, NIAM retains growth inhibitory activity in cells lacking ARF and/or p53, and knockdown experiments revealed that it is not essential for ARF-mediated growth inhibition. Thus, NIAM and ARF act in separate anti-proliferative pathways that intersect mechanistically and suppress growth more effectively when jointly activated. Intriguingly, silencing of NIAM accelerated chromosomal instability, and microarray analyses showed reduced NIAM mRNA expression in numerous primary human tumors. This study identifies a novel protein with tumor suppressor-like behaviors and functional links to ARF-MDM2-p53 signaling. PMID- 17110380 TI - Coordinate suppression of ERBB2 and ERBB3 by enforced expression of micro-RNA miR 125a or miR-125b. AB - Deregulation of micro-RNAs (miRNAs) is emerging as a major aspect of cancer etiology because their capacity to direct the translation and stability of targeted transcripts can dramatically influence cellular physiology. To explore the potential of exogenously applied miRNAs to suppress oncogenic proteins, the ERBB oncogene family was chosen with a bioinformatics search identifying targeting seed sequences for miR-125a and miR-125b within the 3'-untranslated regions of both ERBB2 and ERBB3. Using the human breast cancer cell line SKBR3 as a model for ERBB2 and ERBB3 dependence, infection of these cells with retroviral constructs expressing either miR-125a or miR-125b resulted in suppression of ERBB2 and ERBB3 at both the transcript and protein level. Luciferase constructs containing the 3' 3'-untranslated regions of ERBB2 and ERBB3 demonstrated approximately 35% less activity in miR-125a- and miR-125b-expressing cells relative to controls. Additionally, phosphorylation of ERK1/2 and AKT was suppressed in SKBR3 cells overexpressing either miR-125a or miR-125b. Consistent with suppression of both ERBB2 and ERBB3 signaling, miR-125a-or miR-125b overexpressing SKBR3 cells were impaired in their anchorage-dependent growth and exhibited reduced migration and invasion capacities. Parallel studies performed on MCF10A cells demonstrated that miR-125a or miR-125b overexpression produced only marginal influences on the growth and migration of these non-transformed human mammary epithelial cells. These results illustrate the feasibility of using miRNAs as a therapeutic strategy to suppress oncogene expression and function. PMID- 17110381 TI - The Src homology 3 domain of the beta-subunit of voltage-gated calcium channels promotes endocytosis via dynamin interaction. AB - High voltage-gated calcium channels enable calcium entry into cells in response to membrane depolarization. Association of the auxiliary beta-subunit to the alpha-interaction-domain in the pore-forming alpha1-subunit is required to form functional channels. The beta-subunit belongs to the membrane-associated guanylate kinase class of scaffolding proteins containing a Src homology 3 and a guanylate kinase domain. Although the latter is responsible for the high affinity binding to the alpha-interaction domain, the functional significance of the Src homology 3 domain remains elusive. Here, we show that injection of isolated beta subunit Src homology 3 domain into Xenopus laevis oocytes expressing the alpha1 subunit reduces the number of channels in the plasma membrane. This effect is reverted by coexpressing alpha1 with a dominant-negative mutant of dynamin, a GTPase involved in receptor-mediated endocytosis. Full-length beta-subunit also down-regulates voltage-gated calcium channels but only when lacking the alpha interaction domain. Moreover, isolated Src homology 3 domain and the full-length beta-subunit were found to interact in vitro with dynamin and to internalize the distantly related Shaker potassium channel. These results demonstrate that the beta-subunit regulates the turnover of voltage-gated calcium channels and other proteins in the cell membrane. This effect is mediated by dynamin and depends on the association state of the beta-subunit to the alpha1-pore-forming subunit. Our findings define a novel function for the beta-subunit through its Src homology 3 domain and establish a link between voltage-gated calcium channel activity and the cell endocytic machinery. PMID- 17110382 TI - Metacaspase activity of Arabidopsis thaliana is regulated by S-nitrosylation of a critical cysteine residue. AB - Nitric oxide (NO) regulates a number of signaling functions in both animals and plants under several physiological and pathophysiological conditions. S Nitrosylation linking a nitrosothiol on cysteine residues mediates NO signaling functions of a broad spectrum of mammalian proteins, including caspases, the main effectors of apoptosis. Metacaspases are suggested to be the ancestors of metazoan caspases, and plant metacaspases have previously been shown to be genuine cysteine proteases that autoprocess in a manner similar to that of caspases. We show that S-nitrosylation plays a central role in the regulation of the proteolytic activity of Arabidopsis thaliana metacaspase 9 (AtMC9) and hypothesize that this S-nitrosylation affects the cellular processes in which metacaspases are involved. We found that AtMC9 zymogens are S-nitrosylated at their active site cysteines in vivo and that this posttranslational modification suppresses both AtMC9 autoprocessing and proteolytic activity. However, the mature processed form is not prone to NO inhibition due to the presence of a second S-nitrosylation-insensitive cysteine that can replace the S-nitrosylated cysteine residue within the catalytic center of the processed AtMC9. This cysteine is absent in caspases and paracaspases but is conserved in all reported metacaspases. PMID- 17110383 TI - Expression and functional characterization of the cancer-related serine protease, human tissue kallikrein 14. AB - Human tissue kallikrein 14 (KLK14) is a novel extracellular serine protease. Clinical data link KLK14 expression to several diseases, primarily cancer; however, little is known of its (patho)-physiological role. To functionally characterize KLK14, we expressed and purified recombinant KLK14 in mature and proenzyme forms and determined its expression pattern, specificity, regulation, and in vitro substrates. By using our novel immunoassay, the normal and/or diseased skin, breast, prostate, and ovary contained the highest concentration of KLK14. Serum KLK14 levels were significantly elevated in prostate cancer patients compared with healthy males. KLK14 displayed trypsin-like specificity with high selectivity for P1-Arg over Lys. KLK14 activity could be regulated as follows: 1) by autolytic cleavage leading to enzymatic inactivation; 2) by the inhibitory serpins alpha1-antitrypsin, alpha2-antiplasmin, antithrombin III, and alpha1 antichymotrypsin with second order rate constants (k(+2)/Ki) of 49.8, 23.8, 1.48, and 0.224 microM(-1) min(-1), respectively, as well as plasminogen activator inhibitor-1; and 3) by citrate and zinc ions, which exerted stimulatory and inhibitory effects on KLK14 activity, respectively. We also expanded the in vitro target repertoire of KLK14 to include collagens I-IV, fibronectin, laminin, kininogen, fibrinogen, plasminogen, vitronectin, and insulin-like growth factor binding proteins 2 and 3. Our results indicate that KLK14 may be implicated in several facets of tumor progression, including growth, invasion, and angiogenesis, as well as in arthritic disease via deterioration of cartilage. These findings may have clinical implications for the management of cancer and other disorders in which KLK14 activity is elevated. PMID- 17110384 TI - Conditional stimulation of type V and VI adenylyl cyclases by G protein betagamma subunits. AB - In a yeast two-hybrid screen of mouse brain cDNA library, using the N-terminal region of human type V adenylyl cyclase (hACV) as bait, we identified G protein beta2 subunit as an interacting partner. Additional yeast two-hybrid assays showed that the Gbeta(1) subunit also interacts with the N-terminal segments of hACV and human type VI adenylyl cyclase (hACVI). In vitro adenylyl cyclase (AC) activity assays using membranes of Sf9 cells expressing hACV or hACVI showed that Gbetagamma subunits enhance the activity of these enzymes provided either Galpha(s) or forskolin is present. Deletion of residues 77-151, but not 1-76, in the N-terminal region of hACVI obliterated the ability of Gbetagamma subunits to conditionally stimulate the enzyme. Likewise, activities of the recombinant, engineered, soluble forms of ACV and ACVI, which lack the N termini, were not enhanced by Gbetagamma subunits. Transfection of the C terminus of G protein receptor kinase 2 to sequester endogenous Gbetagamma subunits attenuated the ability of isoproterenol to increase cAMP accumulation in COS-7 cells overexpressing hACVI even when G(i) was inactivated by pertussis toxin. Therefore, we conclude that the N termini of human hACV and hACVI are necessary for interactions with, and regulation by, Gbetagamma subunits both in vitro and in intact cells. Moreover, Gbetagamma subunits derived from a source(s) other than G(i) are necessary for the full activation of hACVI by isoproterenol in intact cells. PMID- 17110385 TI - Open access. PMID- 17110386 TI - Transmission lines, EMF and population mixing. AB - Draper et al. found that the incidence of childhood leukaemia was slightly elevated for children living at distances of 200-600 m from high-voltage transmission lines. This elevation cannot be explained by EMF exposure and it is suggested that it may be due to population mixing in housing developments which followed the construction of the lines. PMID- 17110387 TI - Fluence to organ dose conversion coefficients calculated with the voxel model NORMAN-05 and the MCNPX Monte Carlo code for external monoenergetic photons from 20 keV to 100 MeV. AB - Photons conversion coefficients from 20 keV to 100 MeV have been calculated with the voxel model NORMAN-05 using the MCNPX code. Both kerma approximation and electronic transport were employed and the results compared with published data. In the near future, ICRP group DOCAL will issue a new computational model, based on the GSF-GOLEM, which is intended to be the reference adult male voxel model for ICRP. NORMAN-05 well approximates the western-caucasian standard man characteristics, in terms of body height (176 cm) and mass (73 kg) and masses of the organs. It is not intended to substitute, or to be an alternative, to the future official ICRP voxel model, but thanks to its accuracy and its "standard man structure" can be useful to evaluate the intrinsic uncertainties associated with the dose quantities evaluated adopting different voxel models. For such reason data obtained with NORMAN-05 could be easily compared with those that will be derived from the future ICRP model. PMID- 17110388 TI - Performance of a personal neutron dosemeter based on direct ion storage at workplace fields in the nuclear industry. AB - In the framework of the EVIDOS project, funded by the EC, measurements were carried out using dosemeters, based on ionisation chambers with direct ion storage (DIS-N), at several workplace fields, namely, at a fuel processing plant, a boiling and a pressurised water reactor, and near transport and storage casks. The measurements and results obtained with the DIS-N in these workplaces, which are representative for the nuclear industry, are described in this study. Different dosemeter configurations of converter and shielding materials were considered. The results are compared with values for personal dose equivalent which were assessed within the EVIDOS project by other partners. The advantages and limitations of the DIS-N dosemeter are discussed. PMID- 17110389 TI - Evaluation of dose equivalent by the electronic personal dosemeter for neutron 'Saphydose-N' at different workplaces of nuclear facilities. AB - This paper presents the results of measurements made with the electronic personal neutron Saphydose-N during the four campaigns of the European contract EVIDOS (EValuation of Individual DOSimetry in mixed neutron and photon radiation fields). These measurements were performed at Institute for Radiological Protection and Nuclear Safety (IRSN) in France (C0), at the Krummel Nuclear Power Plant in Germany (C1), at the VENUS Research Reactor and the Belgonucleaire fuel processing plant in Belgium (C2) and at the Ringhals Nuclear Power Plant in Sweden (C3). The results for Saphydose-N are compared with reference values for dose equivalent. PMID- 17110390 TI - Specific absorbed fraction for Korean adult voxel phantom from internal photon source. AB - Absorbed fraction (AF) and specific absorbed fraction (SAF) are crucial values for the calculation of radionuclide S-values and consequently for internal dose estimates. The formalism of the Medical Internal Radiation Dose (MIRD) committee of the Society of Nuclear Medicine (SNM) has been utilised as a standard in the calculation of individual organ doses for biologically distributed radionuclides and for different types of radiation. Although those quantities are highly sensitive to individual anatomical difference, the SAF dataset calculated by Caucasian-based stylised phantoms have been applied to Korean population until now. This study was intended to calculate the SAFs by using realistic Korean voxel phantom and Monte Carlo transport technique for the first time and compare the results with those of the existing Caucasian-based data and the Korean stylised phantom published recently. The up-to-date realistic Korean voxel phantom, KTMAN-2, which was developed from computed tomography (CT) images of an average Korean adult male, was employed for Monte Carlo calculation using EGSnrc user-code, developed for the purpose of this study. The SAFs for 32 target organs and tissues from the photon source, uniformly deposited in a total of 37 source organs and tissues, were calculated from KTMAN-2. The results were compared with those for an adult phantom of Oak Ridge National Laboratory (ORNL) and Korean adult stylised phantom. Two major reasons of discrepancy were analysed: (1) racial difference between the Korean and the Caucasian and (2) anatomical difference between stylised and voxel phantoms. When the source organ was identical to the target organ, difference in SAF caused by the difference in target-organ mass between the Korean and the Caucasian phantoms was mainly observed. When the source and target organs were not identical, significant difference in SAF was observed which was mainly attributed to the difference in inter-organ distance and organ shape between voxel and stylised phantoms. PMID- 17110391 TI - Agrin is required for posterior development and motor axon outgrowth and branching in embryonic zebrafish. AB - Although recent studies have extended our understanding of agrin's function during development, its function in the central nervous system (CNS) is not clearly understood. To address this question, zebrafish agrin was identified and characterized. Zebrafish agrin is expressed in the developing CNS and in nonneural structures such as somites and notochord. In agrin morphant embryos, acetylcholine receptor (AChR) cluster number and size on muscle fibers at the choice point were unaffected, whereas AChR clusters on muscle fibers in the dorsal and ventral regions of the myotome were reduced or absent. Defects in the axon outgrowth by primary motor neurons, subpopulations of branchiomotor neurons, and Rohon-Beard sensory neurons were also observed, which included truncation of axons and increased branching of motor axons. Moreover, agrin morphants exhibit significantly inhibited tail development in a dose-dependent manner, as well as defects in the formation of the midbrain-hindbrain boundary and reduced size of eyes and otic vesicles. Together these results show that agrin plays an important role in both peripheral and CNS development and also modulates posterior development in zebrafish. PMID- 17110392 TI - General practitioners' perceptions of antimicrobial resistance: a qualitative study. AB - OBJECTIVES: Interventions aimed at enhancing the quality of antibiotic prescribing often highlight the threat of antimicrobial resistance. Although most antibiotics are prescribed by general practitioners (GPs), little is known of their perceptions of this issue. The aim of this study was therefore to achieve a deeper understanding of GPs' perceptions of antimicrobial resistance. METHODS: A qualitative interview, grounded theory study. Forty GPs were interviewed, 26 from high fluoroquinolone prescribing practices and 14 from average fluoroquinolone prescribing practices. RESULTS: Most GPs were concerned about the broad issue of antimicrobial resistance and agreed that it was a growing problem. However, many said they infrequently encountered its consequences in their everyday practice and some questioned the evidence linking their prescribing decisions to resistance and poorer outcomes for their patients. They felt conflicted by their apparent inability to influence the problem in the face of many other competing demands. A number said they would welcome more information from their microbiological colleagues about resistance patterns locally, and felt that undergraduate and graduate education about antimicrobial prescribing and resistance should be enhanced. However, a few mentioned that a heightened awareness of antimicrobial resistance locally may cause them to prescribe more second line agents as empirical therapy. CONCLUSIONS: Antimicrobial resistance is only one of a range of important influences on GPs decisions whether or not to prescribe an antibiotic and is not the most immediate. These influences all need to be taken into account when promoting a more cautious use of antibiotics in primary care. More information from microbiologist colleagues about local resistance would be clinically useful, but on its own, may paradoxically influence some GPs to prescribe newer, broader spectrum agents more often. PMID- 17110393 TI - Wide geographic spread of diverse acquired AmpC beta-lactamases among Escherichia coli and Klebsiella spp. in the UK and Ireland. AB - OBJECTIVES: To determine the distribution of acquired AmpC beta-lactamases in 173 isolates of Escherichia coli and Klebsiella spp. submitted to the UK's national reference laboratory for antibiotic resistance. METHODS: MICs were determined and interpreted according to BSAC guidelines. Candidate isolates were those resistant to cefotaxime and/or ceftazidime, irrespective of addition of clavulanic acid. Genes encoding six phylogenetic groups of acquired AmpC enzymes were sought by PCR. Selected isolates were compared by pulsed-field gel electrophoresis (PFGE), and one bla(AmpC) amplicon was sequenced. RESULTS: Genes encoding acquired AmpC enzymes were detected in 67 (49%) candidate E. coli and 21 (55%) Klebsiella spp. Sixty isolates produced CIT-type enzymes, 14 had ACC types, 11 had FOX types and 3 had DHA enzymes. The low-level cephalosporin resistance of the remaining isolates (n = 85; 49%) was inferred to result from reduced permeability or, in E. coli, from hyperexpression of chromosomal ampC. Twenty-four E. coli isolates from one hospital produced a CIT-type enzyme, with 20 of these additionally producing a group 1 CTX-M extended-spectrum beta-lactamase. PFGE indicated that these isolates belonged to UK epidemic strain A, which normally produces CTX-M-15, but no acquired AmpC. Sequencing a representative bla(AmpC) amplicon indicated that in one centre this strain had acquired a novel CMY-2 variant, designated CMY-23. CONCLUSIONS: Diverse acquired AmpC enzymes occur in E. coli and Klebsiella spp. isolates in the UK and Ireland, with CIT types the most common. Producers are geographically scattered, but with some local outbreaks. Acquisition of a CMY-2 like enzyme by E. coli epidemic strain A suggests that these enzymes may be poised to become an important public health issue. PMID- 17110394 TI - Effect of timing and duration of azithromycin therapy of leptospirosis in a hamster model. AB - OBJECTIVES: Azithromycin is not associated with significant adverse effects or restricted usage in certain populations unlike standard antileptospirosis agents. In this study, the utility of short courses of azithromycin in treating or preventing leptospirosis was investigated in a lethal hamster model. METHODS: All hamsters were infected intraperitoneally with 10(5) leptospires. In experiment one, animals received 5 mg/kg of doxycycline or 10 mg/kg of azithromycin via intraperitoneal injection beginning on the second day after infection and continuing once daily for 1, 2, 3 or 5 days. In experiment two, animals received 1 or 2 day courses of azithromycin initiated 2 or 4 days following infection, or 4 days prior to infection. RESULTS: All untreated control animals died between the sixth and ninth day following infection. In experiment one, survival rates in the doxycycline groups were 0, 50, 80 and 100% for those animals treated for 1, 2, 3 and 5 days, respectively. Except for the 1 day treatment group (which had an 80% survival), there was 100% survival in all azithromycin-treated groups. In experiment two, all animals treated after infection survived until study completion. No animals survived with 1 day of therapy started 4 days prior to infection while only 20% survived if they received 2 days. CONCLUSIONS: These results suggest short-course therapy with azithromycin, even started well after infection, is efficacious in preventing mortality from acute leptospirosis. PMID- 17110395 TI - Pattern and rate of indel evolution inferred from whole chloroplast intergenic regions in sugarcane, maize and rice. AB - Microstructural changes such as insertions and deletions (=indels) are a major driving force in the evolution of non-coding DNA sequences. To better understand the mechanisms by which indel mutations arise, as well as the molecular evolution of non-coding regions, the number and pattern of indels and nucleotide substitutions were compared in the whole chloroplast genomes. Comparisons were made for a total of over 38 kb non-coding DNA sequences from 126 intergenic regions in two data sets representing species with different divergence times: sugarcane and maize and Oryza sativa var. indica and japonica. The main findings of this study are: (i) Approximately half of all indels are single nucleotide indels. This observation agrees with previous studies in various organisms. (ii) The distribution and number of indels was different between two data sets, and different patterns were observed for tandem repeat and non-repeat indels. (iii) Distribution pattern of tandem repeat indels showed statistically significant bias towards A/T-rich. (iv) The rate of indel mutation was estimated to be approximately 0.8 +/- 0.04 x 10(-9) per site per year, which was similar to previous estimates in other organisms. (v) The frequencies of nucleotide substitutions and indels were significantly lower in inverted repeat (IR). PMID- 17110396 TI - A randomized clinical trial of clomiphene citrate versus low dose recombinant FSH for ovarian hyperstimulation in intrauterine insemination cycles for unexplained and male subfertility. AB - BACKGROUND: Controlled ovarian hyperstimulation with intrauterine insemination (IUI) is a widely accepted treatment for unexplained and male subfertility. No consensus exists about the drug of first choice to be used as hyperstimulation. This randomized multicentre trial using a parallel design compares the efficacy of clomiphene citrate (CC) with that of recombinant FSH (rFSH). METHODS: Couples with primary unexplained or male subfertility were randomized to receive CC or rFSH for ovarian hyperstimulation. The treatment was continued for up to four cycles unless pregnancy occurred. Cycles with more than three follicles were cancelled. Cumulative pregnancy rates and live birth rates were primary outcomes. Cancellation during treatment and multiple birth rates are secondary outcomes. Results were analysed following the intention-to-treat principle. RESULTS: Seventy couples with male subfertility and 68 couples with unexplained subfertility were included. Seventy-one women received CC, and 67 received rFSH. Twenty-seven pregnancies were observed in the CC group (38%) and 23 in the rFSH group (34.3%) relative risk (RR) 1.11 [95% confidence interval (95% CI) 0.71 1.73]. The live birth rate was 28.2% (20/71) and 26.9% (18/67) for CC and rFSH, respectively, RR 1.05 (95% CI 0.61-1.80). Overall, the live birth rates per cycle were 10% for CC-stimulated and 8.7% for rFSH stimulated cycles. The total multiple pregnancy rate was 6.0%. Thirty-five cycles (8.6%) were cancelled because of four or more follicles (CC, n = 17; rFSH, n = 18). CONCLUSIONS: In couples with primary unexplained or male subfertility participating in an IUI program, ovarian hyperstimulation can be achieved by CC or rFSH. No significant difference in live birth rates between CC and rFSH was observed. Being less expensive, CC seems the more cost-effective drug and therefore, can be offered as drug of first choice. PMID- 17110397 TI - Endocrine profile in serum and follicular fluid differs after ovarian stimulation with HP-hMG or recombinant FSH in IVF patients. AB - BACKGROUND: Highly purified menotrophin (HP-hMG) has been associated with fewer oocytes retrieved and a higher proportion of top-quality embryos compared with recombinant FSH (rFSH). METHODS: A randomized, assessor-blind, multinational trial in 731 women undergoing IVF after stimulation with HP-hMG (MENOPUR) (n = 363) or rFSH (GONAL-F) (n = 368) following a long GnRH agonist protocol was conducted. Blood was collected before, during and after stimulation. Fluid was collected from follicles > or =17 mm. RESULTS: Serum androstenedione, total testosterone and free androgen index (FAI) were higher (P < 0.001) with HP-hMG than with rFSH after starting stimulation. At the end of stimulation, serum estradiol was higher (P = 0.031) with HP-hMG, whereas progesterone was higher (P < 0.001) with rFSH, even after adjusting for ovarian response. Serum LH was not different between treatments. Mean mid- and end-follicular hCG levels in the HP hMG group were 2.5 and 2.9 IU/l, respectively. Follicular fluid levels of FSH, LH, hCG, androstenedione, testosterone, FAI and estradiol and ratios of estradiol:androstenedione, estradiol:total testosterone and estradiol:progesterone were higher (P < 0.001) with HP-hMG, whereas progesterone was higher (P < 0.001) with rFSH. CONCLUSION: Major differences in serum and follicular fluid endocrine profile exist after stimulation with HP-hMG or rFSH. Exogenous LH activity induces a differential endocrine environment influencing oocyte quantity and quality, which may be of relevance for clinical outcome. PMID- 17110398 TI - Fresh human orthotopic ovarian cortex transplantation: long-term results. AB - BACKGROUND: Ovarian orthotopic transplantation in patients with premature ovarian failure is reported to result in full-term pregnancies. Ischaemia and freezing/thawing are potentially injurious for tissues. This study was designed to analyse the effect of ischaemia on long-term ovarian function in humans. METHODS: Prospective case-control study. Subjects were 12 premenopausal women undergoing hysterectomy and fresh orthotopic transplantation of the entire ovarian cortex plus a control group of five patients undergoing hysterectomy only. Follow-up lasted 2 years. Serum FSH and anti-Mullerian hormone (AMH) were recorded, and ovulatory cycles were determined by vaginal ultrasound and serum progesterone levels. RESULTS: Follow-up showed that ovulation was restored in 11 of the 12 patients who received grafts over the duration of the study (9.3 +/- 1.73 ovulations versus 12.0 +/- 0.86 in controls, NS), and 9 of 12 patients remained ovulatory after 2 years. We identified four patterns of FSH secretion during the study, 5 of 12 (41.7%) women having the same pattern as controls. There was a trend for serum AMH levels 7 days after surgery (0.16 +/- 0.02 microg/l) to be lower than pre-surgery levels (0.38 +/- 0.09 microg/l, P = 0.07) and higher in women whose FSH patterns suggested normal ovarian function, but the results did not reach significance. After transplantation, FSH correlated more closely (r = -0.639, P = 0.02) with normal ovarian function than AMH (r = 0.465, P = 0.12). CONCLUSIONS: Fresh orthotopic ovarian cortex transplantation is a viable procedure. It maintains normal ovarian function after 2 years in 75% of cases and preserves ovarian function against ischaemia in 41.7% of patients. PMID- 17110399 TI - Testis/sperm-specific histone 2B in the sperm of donors and subfertile patients: variability and relation to chromatin packaging. AB - BACKGROUND: The compaction of human sperm chromatin is the result of replacement of approximately 85% of histones with protamines. Germ-line testis/sperm-specific histone 2B (TSH2B) has been detected in only approximately 30% of mature spermatozoa. Its level in the semen of subfertile patients varies; its function is unknown. We evaluated TSH2B in the sperm samples of 23 donors and 49 subfertile patients and assessed its association with chromatin compaction status. METHODS: TSH2B level was measured using immunoblotting. Chromatin packaging quality was evaluated by staining with chromomycin A3 (CMA3) which marked spermatozoa with defective packaging. To assess both TSH2B and chromatin status in the same spermatozoon, CMA3 staining and TSH2B immunolocalization were performed sequentially. RESULTS: A significant correlation (r = 0.55, P = 0.0027) was found between TSH2B level and percentage of CMA3-positive sperm in patient and donor semen samples. When individual spermatozoa were assessed for these parameters, 92% of TSH2B-containing cells were also CMA3 positive. Variation in the total sperm TSH2B level was less in donors than in patients. CONCLUSIONS: CMA3 positive staining of TSH2B-containing individual spermatozoa and a significant correlation between the total TSH2B level and CMA3 percentage in semen samples suggest a structural role for TSH2B in sperm chromatin organization. Low variability of TSH2B level in donors implies a mechanism (however unknown) regulating this parameter. PMID- 17110400 TI - Social support during pregnancy: effects on maternal depressive symptoms, smoking and pregnancy outcome. AB - BACKGROUND: The goal was to study the effects of social support during pregnancy on maternal depressive symptoms, quality of life and pregnancy outcomes. METHODS: Eight hundred ninety-six women were prospectively studied in the first trimester of pregnancy and following completion of the pregnancy. The sample was divided into quartiles yielding groups of low, medium and high social support based on perceived social support. RESULTS: Pregnant women with low support reported increased depressive symptoms and reduced quality of life. The effects of social support on pregnancy outcomes were particularly pronounced in women who had smoked during pregnancy, with significant main effects of social support in a two way analysis of variance (smoking status and social support) for child body length (F = 4.26, P = 0.04; 50.43 +/- 2.81 cm with low support versus 51.76 +/- 2.31 cm with high support) and birthweight (F = 11.35, P = 0.001; 3175 +/- 453 g with low support versus 3571 +/- 409 g with high support). In smokers, pregnancy complications occurred more frequently when given low support {34 versus 10.3% with high support, chi(2) = 5.49, P = 0.019; relative risk (RR) = 3.3 [95% confidence interval (95% CI) = 1.1-10.2]}, and the proportion of preterm deliveries was greater given low support (10.0 versus 0% with high support, chi(2) = 3.84, P = 0.05, odds ratio = 8.1). CONCLUSIONS: Lack of social support constitutes an important risk factor for maternal well-being during pregnancy and has adverse effects on pregnancy outcomes. PMID- 17110401 TI - Implications for molecular mechanisms of glycoprotein hormone receptors using a new sequence-structure-function analysis resource. AB - Comparison between wild-type and mutated glycoprotein hormone receptors (GPHRs), TSH receptor, FSH receptor, and LH-chorionic gonadotropin receptor is established to identify determinants involved in molecular activation mechanism. The basic aims of the current work are 1) the discrimination of receptor phenotypes according to the differences between activity states they represent, 2) the assignment of classified phenotypes to three-dimensional structural positions to reveal 3) functional-structural hot spots and 4) interrelations between determinants that are responsible for corresponding activity states. Because it is hard to survey the vast amount of pathogenic and site-directed mutations at GPHRs and to improve an almost isolated consideration of individual point mutations, we present a system for systematic and diversified sequence-structure function analysis (http://www.fmp-berlin.de/ssfa). To combine all mutagenesis data into one set, we converted the functional data into unified scaled values. This at least enables their comparison in a rough classification manner. In this study we describe the compiled data set and a wide spectrum of functions for user driven searches and classification of receptor functionalities such as cell surface expression, maximum of hormone binding capability, and basal as well as hormone-induced Galphas/Galphaq mediated cAMP/inositol phosphate accumulation. Complementary to known databases, our data set and bioinformatics tools allow functional and biochemical specificities to be linked with spatial features to reveal concealed structure-function relationships by a semiquantitative analysis. A comprehensive discrimination of specificities of pathogenic mutations and in vitro mutant phenotypes and their relation to signaling mechanisms of GPHRs demonstrates the utility of sequence-structure-function analysis. Moreover, new interrelations of determinants important for selective G protein-mediated activation of GPHRs are resumed. PMID- 17110402 TI - The challenge of explaining why quality improvement has not done better. PMID- 17110403 TI - The long-term cost-effectiveness of cardiac resynchronization therapy with or without an implantable cardioverter-defibrillator. AB - AIMS: Cardiac resynchronization therapy (CRT-P) is an effective treatment for patients with heart failure and cardiac dyssynchrony with moderate or severe symptoms despite pharmacological therapy. The addition of an implantable cardioverter-defibrillator (ICD) function may further reduce the risk of sudden death. We assessed the cost-effectiveness of CRT-P compared with medical therapy (MT) alone, and the cost-effectiveness of CRT-ICD + MT compared with CRT-P + MT, on incremental cost per quality adjusted life year (QALY) and life year using data from two landmark clinical trials. METHODS AND RESULTS: A Markov model with Monte Carlo simulation to assess costs, life years, and QALYs associated with CRT (+/- ICD) and MT in patients with heart failure and cardiac dyssynchrony, on the basis of a UK healthcare perspective was constructed. NYHA class distribution and transitions, associated health utilities, rates and cause of hospitalization and death were estimated from individual patient data from the CArdiac REsychronization in Heart Failure (CARE-HF trial). The estimated additional benefit on survival of an ICD was based on results from COMPANION. The base case analysis used 10 000 individual life-time simulations assuming a battery life of 6 years for CRT-P and 7 years for CRT-ICD. From a life-time perspective in a 65 year-old patient, the incremental cost-effectiveness of CRT-P compared with MT is 7538 euros (95% CI 5325-11,784 euros) per QALY gained and 7011euros (95% CI 5346 10,003 euros) per life year gained. The incremental cost-effectiveness of CRT-ICD compared with CRT-P is 47,909 euros (95% CI 35,703-79,438 euros) per QALY gained, and 35,864 euros (95% CI 26,709-56,353 euros) per life year gained. CONCLUSION: Long-term treatment with CRT-P appears cost-effective compared with MT alone. From a life-time perspective, assuming a reasonable life expectancy when receiving effective treatment for heart failure, CRT-ICD may also be considered cost-effective when compared with CRT-P + MT. PMID- 17110404 TI - Sequential autologous peripheral blood stem cell transplantation and kidney transplantaion of light chain deposition disease. PMID- 17110405 TI - Renal handling of cystatin C. PMID- 17110406 TI - Chloride and the endosomal-lysosomal pathway: emerging roles of CLC chloride transporters. AB - Several members of the CLC family of Cl- channels and transporters are expressed in vesicles of the endocytotic-lysosomal pathway, all of which are acidified by V type proton pumps. These CLC proteins are thought to facilitate vesicular acidification by neutralizing the electric current of the H+-ATPase. Indeed, the disruption of ClC-5 impaired the acidification of endosomes, and the knock-out (KO) of ClC-3 that of endosomes and synaptic vesicles. KO mice are available for all vesicular CLCs (ClC-3 to ClC-7), and ClC-5 and ClC-7, as well as its beta subunit Ostm1, are mutated in human disease. The associated mouse and human pathologies, ranging from impaired endocytosis and nephrolithiasis (ClC-5) to neurodegeneration (ClC-3), lysosomal storage disease (ClC-6, ClC-7/Ostm1) and osteopetrosis (ClC-7/Ostm1), were crucial in identifying the physiological roles of vesicular CLCs. Whereas the intracellular localization of ClC-6 and ClC 7/Ostm1 precluded biophysical studies, the partial expression of ClC-4 and -5 at the cell surface allowed the detection of strongly outwardly rectifying currents that depended on anions and pH. Surprisingly, ClC-4 and ClC-5 (and probably ClC 3) do not function as Cl- channels, but rather as electrogenic Cl--H+ exchangers. This hints at an important role for luminal chloride in the endosomal-lysosomal system. PMID- 17110407 TI - Autonomic/metabolic interactions modulating the exercise pressor reflex: the purinergic hypothesis. PMID- 17110408 TI - Entia non sunt multiplicanda praeter necessitatem. PMID- 17110409 TI - Central command and the onset of exercise. PMID- 17110410 TI - Propagation of postsynaptic currents and potentials via gap junctions in GABAergic networks of the rat hippocampus. AB - The integration of synaptic signalling in the mammalian hippocampus underlies higher cognitive functions such as learning and memory. We have studied the gap junction-mediated cell-to-cell and network propagation of GABA(A) receptor mediated events in stratum lacunosum moleculare interneurons of the rat hippocampus. Propagated events were identified both in voltage- and current-clamp configurations. After blockade of ionotropic excitatory synaptic transmission, voltage-clamp recordings with chloride-loaded electrodes (predicted GABA(A) receptor reversal potential: 0 mV) at -15 mV revealed the unexpected presence of spontaneous events of opposite polarities. Inward events were larger and kinetically faster when compared to outward currents. Both types of events were blocked by gabazine, but only outward currents were significantly affected by the gap junction blocker carbenoxolone, indicating that outward events originated in electrically coupled neurons. These results were in agreement with computational modelling showing that propagated events were modulated in size and shape by their relative distance to the gap junction site. Paired recordings from electrically coupled interneurons performed with high- and low-chloride pipettes (predicted GABA(A) receptor reversal potentials: 0 mV and -80 mV, respectively) directly demonstrated that depolarizing postsynaptic events could propagate to the cell recorded with the low-chloride solution. Cell-to-cell propagation was abolished by carbenoxolone, and was not observed in uncoupled pairs. Application of 4-aminopyridine on slices resulted in spontaneous network activation of interneurons, which was driven by excitatory GABA(A) receptor-mediated input. Population activity was greatly depressed by carbenoxolone, suggesting that propagation of depolarizing synaptic GABAergic potentials may be a critical determinant of interneuronal synchronous bursting in the hippocampus. PMID- 17110411 TI - Manganese stimulates luteinizing hormone releasing hormone secretion in prepubertal female rats: hypothalamic site and mechanism of action. AB - We have shown recently that Mn2+ stimulates gonadotropin secretion via an action at the hypothalamic level, and a diet supplemented with a low dose of the element is capable of advancing the time of female puberty. In this study, we used an in vitro approach to investigate the mechanism by which Mn2+ induces luteinizing hormone-releasing hormone (LHRH) secretion from prepubertal female rats. The medial basal hypothalamus from 30-day-old rats was incubated in Locke solution for 30 min to assess basal LHRH secretion, then incubated with buffer alone or buffer plus either a nitric oxide synthase (NOS) inhibitor (N-monomethyl-L arginine (NMMA); 300 or 500 microM) or a soluble guanylyl cyclase (sGC) inhibitor (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ); 100 or 250 microM) for another 30 min. Finally, the incubation continued for a further 30 min, but in the presence of MnCl2 (50 or 250 microM) to assess the effect of the blockers on stimulated LHRH secretion. Both 50 and 250 microM MnCl2 stimulated LHRH release (P < 0.05 and P < 0.01, respectively). The addition of 300-500 microM NMMA to the medium did not block Mn2+-stimulated release of LHRH, even with the higher dose of MnCl2. Furthermore, while 50, 100 and 250 microM MnCl2 all significantly induced LHRH release, the two lowest doses did not stimulate total nitrite released from the same tissue, an effect only observed with the highest dose. Taken together, these data suggest that Mn2+ is not an effective stimulator of NO. Conversely, inhibiting sGC with ODQ blocked the Mn2+-stimulated secretion of LHRH in a dose-dependent manner, indicating that GC is the site of action of Mn2+. Additionally, we showed that Mn2+ stimulated cGMP and LHRH from the same tissues, and that downstream blocking of protein kinase G formation with KT5823 (10 microM) inhibited Mn2+-induced LHRH release. These data demonstrate that the principal action of Mn2+ within the hypothalamus is to activate sGC directly and/or as a cofactor with available NO, hence generating cGMP and resulting in prepubertal LHRH release. PMID- 17110412 TI - Dispersion of repolarization and refractoriness are determinants of arrhythmia phenotype in transgenic mice with long QT. AB - Enhanced dispersion of repolarization (DR) and refractoriness may be a unifying mechanism central to arrhythmia genesis in the long QT (LQT) syndrome. The role of DR in promoting arrhythmias was investigated in several strains of molecularly engineered mice: (a) Kv4.2 dominant negative transgenic (Kv4.2DN) that lacks the fast component of the transient outward current, I(to,f), have action potential (AP) and QT prolongation, but no spontaneous arrhythmias, (b) Kv1.4 targeted mice (Kv1.4-/-) that lack the slow component of I(to) (I(to,s)), have no QT prolongation and no spontaneous arrhythmias, and (c) double transgenic (Kv4.2DN x Kv1.4-/-) mice that lack both I(to,f) and I(to,s), have AP and QT prolongation, and spontaneous ventricular tachyarrhythmias. Hearts were perfused, stained with di-4-ANEPPS and optically mapped. Activation patterns and conduction velocities were similar between the strains but AP duration at 75% recovery (APD75) was longer in Kv4.2DN (28.0 +/- 2.5 ms, P < 0.01, n = 6), Kv1.4-/- (28.4 +/- 0.4 ms, P < 0.01, n = 5) and Kv4.2DN x Kv1.4-/- (34.3 +/- 2.6 ms, P < 0.01, n = 6) mice than controls (20.3 +/- 1.0 ms, n = 5). Dispersion of refractoriness between apex and base was markedly reduced in Kv4.2DN (0.3 +/- 0.5 ms, n = 6, P < 0.05) but enhanced in Kv1.4-/- (14.2 +/- 2.0 ms, n = 5, P < 0.05) and Kv4.2DN x Kv1.4(-/-) (15.0 +/- 3 ms, n = 5, P < 0.5) mice compared with controls (10 +/- 2 ms, n = 5). A premature pulse elicited ventricular tachycardia (VT) in Kv1.4-/- (n = 4/5) and Kv4.2DN x Kv1.4-/- hearts (n = 5/5) but not Kv4.2DN hearts (n = 0/6). Voltage clamp recordings showed that I(to,f) was 30% greater in myocytes from the apex than base which may account for the absence of DR in Kv4.2DN mice. Thus, dispersion of repolarization (DR) appears to be an important determinant of arrhythmia vulnerability. PMID- 17110413 TI - The effects of acute oral antioxidants on diving-induced alterations in human cardiovascular function. AB - Diving-induced acute alterations in cardiovascular function such as arterial endothelial dysfunction, increased pulmonary artery pressure (PAP) and reduced heart function have been recently reported. We tested the effects of acute antioxidants on arterial endothelial function, PAP and heart function before and after a field dive. Vitamins C (2 g) and E (400 IU) were given to subjects 2 h before a second dive (protocol 1) and in a placebo-controlled crossover study design (protocol 2). Seven experienced divers performed open sea dives to 30 msw with standard decompression in a non-randomized protocol, and six of them participated in a randomized trial. Before and after the dives ventricular volumes and function and pulmonary and brachial artery function were assessed by ultrasound. The control dive resulted in a significant reduction in flow-mediated dilatation (FMD) and heart function with increased mean PAP. Twenty-four hours after the control dive FMD was still reduced 37% below baseline (8.1 versus 5.1%, P = 0.005), while right ventricle ejection fraction (RV-EF), left ventricle EF and endocardial fractional shortening were reduced much less (approximately 2 3%). At the same time RV end-systolic volume was increased by 9% and mean PAP by 5%. Acute antioxidants significantly attenuated only the reduction in FMD post dive (P < 0.001), while changes in pulmonary artery and heart function were unaffected by antioxidant ingestion. These findings were confirmed by repeating the experiments in a randomized study design. FMD returned to baseline values 72 h after the dive with pre-dive placebo, whereas for most cardiovascular parameters this occurred earlier (24-48 h). Right ventricular dysfunction and increased PAP lasted longer. Acute antioxidants attenuated arterial endothelial dysfunction after diving, while reduction in heart and pulmonary artery function were unchanged. Cardiovascular changes after diving are not fully reversed up to 3 days after a dive, suggesting longer lasting negative effects. PMID- 17110414 TI - Effects of L-type Ca2+ channel antagonism on ventricular arrhythmogenesis in murine hearts containing a modification in the Scn5a gene modelling human long QT syndrome 3. AB - Ventricular arrhythmogenesis in long QT 3 syndrome (LQT3) involves both triggered activity and re-entrant excitation arising from delayed ventricular repolarization. Effects of specific L-type Ca2+ channel antagonism were explored in a gain-of-function murine LQT3 model produced by a DeltaKPQ 1505-1507 deletion in the SCN5A gene. Monophasic action potentials (MAPs) were recorded from epicardial and endocardial surfaces of intact, Langendorff-perfused Scn5a+/Delta hearts. In untreated Scn5a+/Delta hearts, epicardial action potential duration at 90% repolarization (APD90) was 60.0 +/- 0.9 ms compared with 46.9 +/- 1.6 ms in untreated wild-type (WT) hearts (P < 0.05; n = 5). The corresponding endocardial APD(90) values were 52.0 +/- 0.7 ms and 53.7 +/- 1.6 ms in Scn5a+/Delta and WT hearts, respectively (P > 0.05; n = 5). Epicardial early afterdepolarizations (EADs), often accompanied by spontaneous ventricular tachycardia (VT), occurred in 100% of MAPs from Scn5a+/Delta but not in any WT hearts (n = 10). However, EAD occurrence was reduced to 62 +/- 7.1%, 44 +/- 9.7%, 10 +/- 10% and 0% of MAPs following perfusion with 10 nm, 100 nm, 300 nm and 1 mum nifedipine, respectively (P < 0.05; n = 5), giving an effective IC50 concentration of 79.3 nm. Programmed electrical stimulation (PES) induced VT in all five Scn5a+/Delta hearts (n = 5) but not in any WT hearts (n = 5). However, repeat PES induced VT in 3, 2, 2 and 0 out of 5 Scn5a+/Delta hearts following perfusion with 10 nm, 100 nm, 300 nm and 1 mum nifedipine, respectively. Patch clamp studies in isolated ventricular myocytes from Scn5a+/Delta and WT hearts confirmed that nifedipine (300 nm) completely suppressed the inward Ca2+ current but had no effect on inward Na+ currents. No significant effects were seen on epicardial APD90, endocardial APD90 or ventricular effective refractory period in Scn5a+/Delta and WT hearts following perfusion with nifedipine at 1 nm, 10 nm, 100 nm, 300 nm and 1 microm nifedipine concentrations. We conclude that L-type Ca2+ channel antagonism thus exerts specific anti-arrhythmic effects in Scn5a+/Delta hearts through suppression of EADs. PMID- 17110415 TI - Electroencephalographic evidence for pre-motor cortex activation during inspiratory loading in humans. AB - Faced with mechanical inspiratory loading, awake animals and anaesthetized humans develop alveolar hypoventilation, whereas awake humans do defend ventilation. This points to a suprapontine compensatory mechanism instead of or in addition to the 'traditional' brainstem respiratory regulation. This study assesses the role of the cortical pre-motor representation of inspiratory muscles in this behaviour. Ten healthy subjects (age 19-34 years, three men) were studied during quiet breathing, CO2-stimulated breathing, inspiratory resistive loading, inspiratory threshold loading, and during self-paced voluntary sniffs. Pre triggered ensemble averaging of Cz EEG epochs starting 2.5 s before the onset of inspiration was used to look for pre-motor activity. Pre-motor potentials were present during voluntary sniffs in all subjects (average latency (+/-s.d.): 1325 +/- 521 ms), but also during inspiratory threshold loading (1427 +/- 537 ms) and during inspiratory resistive loading (1109 +/- 465 ms). Pre-motor potentials were systematically followed by motor potentials during inspiratory loading. Pre-motor potentials were lacking during quiet breathing (except in one case) and during CO2-stimulated breathing (except in two cases). The same pattern was observed during repeated experiments at an interval of several weeks in a subset of three subjects. The behavioural component of inspiratory loading compensation in awake humans could thus depend on higher cortical motor areas. Demonstrating a similar role of the cerebral cortex in the compensation of disease-related inspiratory loads (e.g. asthma attacks) would have important pathophysiological implications: it could for example contribute to explain why sleep is both altered and deleterious in such situations. PMID- 17110416 TI - Metabotropic suppression of excitation in murine autaptic hippocampal neurons. AB - Depolarization-induced suppression of excitation (DSE) and inhibition (DSI) are forms of short-term neuronal plasticity involving postsynaptic release of an endocannabinoid and the activation of presynaptic cannabinoid CB1 receptors. We have recently reported that CB1-dependent DSE can be elicited in autaptic cultures of excitatory hippocampal neurons of the mouse. We now report that the same preparation exhibits a parallel G(q)-coupled receptor-dependent production of endocannabinoids causing retrograde inhibition, also via CB1 receptors, which we will refer to as metabotropic suppression of excitation (MSE). We tested a spectrum of G(q)-coupled receptor agonists and found that both muscarinic and metabotropic glutamate receptors (group I) mediate retrograde inhibition via CB1 receptors in autaptic hippocampal neurons. Thus these neurons possess not only the pre- and postsynaptic machinery necessary for DSE but also that for MSE. This permitted a closer examination of MSE and its interaction with other aspects of the endocannabinoid retrograde signalling machinery: MSE mimics and occludes DSE and is itself occluded by the endocannabinoid 2-arachidonoyl glycerol (2-AG), consistent with 2-AG as a likely mediator of MSE. In contrast to DSE, MSE undergoes heterologous desensitization over the time course of minutes. In keeping with data reported for metabotropic suppression of inhibition (MSI) and DSI in the hippocampus, subthreshold MSE and DSE act synergistically. We additionally found that Delta9-tetrahydrocannabinol, which has been shown to attenuate DSE, antagonizes MSE. Finally, we have distinguished a neuronal subpopulation that exhibits DSE and a differential complement of MSE-mediating Gq coupled receptors, making possible contrasting studies of MSE. Autaptic endocannabinoid signalling is rich, robust and complex in a deceptively simple package, including a previously unreported postsynaptic mechanism of adaptation in addition to known presynaptic CB1 desensitization. These adaptive sites offer novel targets for modulation of endogenous cannabinoid signalling. PMID- 17110417 TI - Short- and long-term depression of rat cerebellar parallel fibre synaptic transmission mediated by synaptic crosstalk. AB - Cerebellar granule cell to Purkinje cell synapses have been reported to show plasticity when stimulating the parallel fibres, but not when granule cell axons are stimulated in the granular layer. The latter absence of plasticity has been attributed either to the synapses made by ascending granule cell axons lacking some feature needed to evoke plasticity, such as metabotropic glutamate receptors, or to spillover of glutamate between adjacent active synapses being essential for plasticity to occur and having a greater effect for parallel fibre stimulation than for granular layer stimulation. Here we show that both long-term depression (LTD) and endocannabinoid plasticity can depend on interaction between adjacent synapses. These results focus attention on the need to characterize the spatial pattern of parallel fibre activity evoked by physiological stimuli, in order to assess the conditions under which synaptic plasticity will occur in vivo. PMID- 17110419 TI - Calcitonin gene-related peptide inhibits early B cell development in vivo. AB - Recent in vitro studies suggest that calcitonin gene-related peptide (CGRP) inhibits early B cell differentiation; however, there is no evidence in the intact animal for a role for CGRP in B cell development. Here, we show that in vivo treatment of mice with CGRP reduces the number of IL-7 responsive B cell progenitors in bone marrow. A single CGRP treatment reduces IL-7-responsive B cell progenitors by up to 40% for up to 72 h. The reduction is dose-dependent and can be blocked by a CGRP receptor antagonist, CGRP(8-37). CGRP in serum following injection is highly elevated at 30 min but returns to basal levels by 4 h, suggesting that a single injection of CGRP has long-lasting effects on B cell development. This report provides the first direct in vivo evidence that CGRP, a neuropeptide with multiple effects on mature lymphocytes, also plays a regulatory role in early B cell development in the bone marrow. PMID- 17110418 TI - CXCL1/KC and CXCL5/LIX are selectively produced by corneal fibroblasts and mediate neutrophil infiltration to the corneal stroma in LPS keratitis. AB - The severity of corneal inflammation depends on the activity of infiltrating neutrophils responding to chemotactic factors such as CXC chemokines. This study examines the relative contribution of CXCL1/keratinocyte-derived chemokine (KC), CXCL2/monocyte-inhibitory protein-2 (MIP-2), and CXCL5/LPS-induced chemokine (LIX) in neutrophil recruitment to the corneal stroma during LPS keratitis, where neutrophils infiltrate the corneal stroma at 6 h after LPS injection and peak at 24 h. Consistent with this timeframe, KC was detected after 3 h, reached peak levels at 24 h, and decreased thereafter. In contrast, LIX production was not detected until 8 h after injection and peaked at 24 h. MIP-2 was detected at 3 h but did not reach the levels of KC and LIX. Cell types associated with corneal inflammation produced markedly different chemokines in vitro: Murine corneal fibroblasts (MK/T-1) produced LIX and KC in response to LPS but did not produce MIP-2, whereas peritoneal macrophages and neutrophils produced MIP-2 and KC but did not produce LIX. To determine the role of these chemokines in neutrophil recruitment to the cornea, anti-LIX, anti-KC, or anti-MIP-2 was injected into the corneal stroma of enhanced GFP chimeric mice prior to LPS, and total cell and neutrophil infiltration was examined. Antibody to LIX and KC, injected individually or in combination, significantly inhibited neutrophil recruitment to the cornea, whereas anti-MIP-2 had no inhibitory effect. Together, these findings demonstrate cell-specific production of CXC chemokines and show that LIX and KC mediate neutrophil recruitment into the cornea during LPS keratitis. PMID- 17110420 TI - Characterization of CD8+ T cell repertoire in identical twins discordant and concordant for multiple sclerosis. AB - Autoreactive CD4+ and CD8+ T cells directed against CNS autoantigens may play a role in the development of multiple sclerosis (MS). Identical twins share the same genetic background but not the TCR repertoire that is shaped by the encounter with self or foreign antigens. To gain insights into the interplay between MS and T cell repertoire, peripheral blood CD4+ and CD8+ T lymphocytes and their CCR7+/CCR7- subsets from five pairs of identical twins (four discordant and one concordant for MS; none of which had taken disease-modifying therapy) were compared by TCR beta-chain (TCRB) complementary-determining region 3 (CDR3) spectratyping. CD4+ T cells generally showed a Gaussian distribution, whereas CD8+ T cells exhibited subject-specific, widely skewed TCR spectratypes. There was no correlation between CD8+ T cell oligoclonality and disease. Sequencing of predominant spectratype expansions revealed shared TCRB-CDR3 motifs when comparing inter- and/or intrapair twin members. In many cases, these sequences were homologous to published TCRs, specific for viruses implicated in MS pathogenesis, CNS autoantigens, or copaxone [glatiramer acetate (GA)], implying the occurrence of naturally GA-responding CD8+ T cells. It is notable that these expanded T cell clones with putative pathogenic or regulatory properties were present in the affected as well as in the healthy subject, thus suggesting the existence of a "MS predisposing trait" shared by co-twins discordant for MS. PMID- 17110421 TI - Protein kinase Czeta is required for oleic acid-induced secretion of glucagon like peptide-1 by intestinal endocrine L cells. AB - Long-chain, monounsaturated fatty acids (FAs) stimulate secretion of the incretin hormone, glucagon-like peptide-1 (GLP-1) from the intestinal L cell. Because the atypical protein kinase C (PKC), PKCzeta, is involved in FA signaling in many cells, the role of PKCzeta in FA-induced GLP-1 secretion was investigated, using the murine GLUTag L cell line and primary rat intestinal L cells. GLUTag cells expressed mRNA for several PKC isoforms, including PKCzeta, and PKCzeta protein was localized throughout the cytoplasm in GLUTag and primary L cells as well as normal mouse and rat L cells. Treatment with oleic acid (150-1000 microm) for 2 h increased GLP-1 secretion (P < 0.001), and this was abrogated by the PKCzeta inhibitor ZI (P < 0.05) and PKCzeta small interfering RNA transfection (P < 0.05) but not inhibition of classical/novel PKC isoforms. Although most PKCzeta was localized in the particulate compartment of GLUTag cells, oleate treatment did not alter PKCzeta levels or activity in this cell fraction. GLUTag cells expressed mRNA for the Gq-coupled FA receptor GPR120; however, oleic acid did not induce any changes in Akt, MAPK, or calcium, and pretreatment with LY294002 and PD98059 to inhibit phosphatidylinositol 3-kinase and MAPK, respectively, did not prevent the effects of oleic acid. Finally, GLUTag cells also released GLP-1 in response to arachidonic acid (P < 0.001) but were not affected by other long chain FAs. These findings demonstrate that PKCzeta is required for oleic acid induced GLP-1 secretion. This enzyme may therefore serve as a therapeutic target to enhance GLP-1 release in type 2 diabetes. PMID- 17110422 TI - Analysis of the gene regulatory program induced by the homeobox transcription factor distal-less 3 in mouse placenta. AB - Dlx3, a homeodomain transcription factor, is essential for placental development in the mouse. The Dlx3(-/-) mouse embryo dies at embryonic d 9.5-10 putatively due to placental failure. To develop a more comprehensive understanding of the gene profile regulated by Dlx3, microarray analysis was used to determine differences in gene expression within the placenta of Dlx3(+/+) and Dlx3(-/-) mice. Array analysis revealed differential expression of 401 genes, 33 genes in which signal to log ratio values of null/wild-type were lower than -0.5 or higher than 0.5. To corroborate these findings, quantitative real-time PCR was used to confirm differential expression for 11 genes, nine of which displayed reduced expression and two with enhanced expression in the Dlx3(-/-) mouse. Loss of Dlx3 resulted in a marked reduction (>60%) in mRNA expression of placental growth factor (Pgf), a member of the vascular endothelial growth factor family. Consistent with these results, Pgf secretion from placental explants tended to be reduced in the Dlx3(-/-) mice, compared with wild type. To investigate mechanisms of Dlx3 regulation of Pgf gene transcription, we cloned 5.2 kb of the Pgf 5' flanking sequence for use in reporter gene assays. Expression of the Pgf promoter luciferase reporter containing at least three Dlx3 binding sites was increased markedly by overexpression of Dlx3 supporting the conclusion that Dlx3 may have a direct effect on Pgf promoter activity. These studies provide a novel view of the transcriptome regulated by Dlx3 in mouse placenta. Dlx3 is specifically required for full expression and secretion of Pgf in vivo. Moreover, in vitro studies support the conclusion that Dlx3 is sufficient to directly modulate expression of the Pgf gene promoter in placental cells. PMID- 17110423 TI - Combination therapy with the advanced glycation end product cross-link breaker, alagebrium, and angiotensin converting enzyme inhibitors in diabetes: synergy or redundancy? AB - Blockade of advanced glycation end product (AGE) accumulation with alagebrium with concomitant angiotensin converting enzyme inhibition was tested for effects on renal function and on other postulated mediators of diabetic renal disease including the renin-angiotensin system, AGEs, mitochondrial and cytosolic oxidative stress, and intracellular signaling molecules. Sprague Dawley rats were rendered diabetic with streptozocin and followed consecutively for 32 wk with nondiabetic controls. Groups were treated with ramipril (1 mg/kg.d; wk 0-32); alagebrium (10 mg/kg.d; wk 16-32); or a combination of both. Although individual treatments had significant effects on albuminuria, no further improvements were seen with combination therapy. Changes in urinary vascular endothelial growth factor excretion mirrored those seen in albuminuria. Diabetes was associated with suppression of circulating angiotensin II in the context of increased circulating and renal levels of the AGE, carboxymethyllysine. All treatments attenuated circulating but not renal carboxymethyllysine levels. The renal gene expression of AGE receptor 1 and soluble receptor for advanced glycation end products were markedly reduced by diabetes and normalized with alagebrium. Diabetes induced renal mitochondrial oxidative stress, which was reduced with alagebrium. In the cytosol, both therapies were equally effective in reducing reactive oxygen species production. Increases in membranous protein kinase C activity in diabetes were attenuated by all treatments, whereas diabetes-associated increases in nuclear factor-kappaB p65 translocation remained unaltered by any therapy. It is evident that renin-angiotensin system blockade and AGE inhibition have specific effects. However, many of their downstream effects appear to be similar, suggesting that their renoprotective benefits may ultimately involve common pathways and key points of convergence, which could be important targets for new therapies in diabetic nephropathy. PMID- 17110424 TI - Intracoronary ghrelin infusion decreases coronary blood flow in anesthetized pigs. AB - The peptide ghrelin has been linked to the atherosclerotic process and coronary artery disease. We planned to study, for the first time, the primary effects of ghrelin on the intact coronary circulation and determine the mechanisms involved. In 24 sodium pentobarbitone-anesthetized pigs, changes in anterior descending coronary blood flow caused by intracoronary infusion of ghrelin at constant heart rate and arterial pressure were assessed using electromagnetic flowmeters. In 20 pigs, intracoronary infusion of ghrelin decreased coronary blood flow without affecting left ventricular maximum rate of change of left ventricular systolic pressure (dP/dt(max)), filling pressures of the heart or plasma levels of GH. In four pigs, this decrease was graded by step increments of infused dose of the hormone. The mechanisms of the above response were studied in the 20 pigs by repeating the experiment after coronary flow had returned to the control values observed before infusion. The ghrelin-induced coronary vasoconstriction was not affected by iv atropine (five pigs) or phentolamine (five pigs). This response was abolished by iv butoxamine (five pigs) and intracoronary N(omega)-nitro-l arginine methyl ester (five pigs), even after reversing the increase in arterial pressure and coronary vascular resistance caused by the two blocking agents with iv infusion of papaverine. The present study showed that intracoronary infusion of ghrelin primarily caused coronary vasoconstriction. The mechanisms of this response were shown to involve the inhibition of a vasodilatory beta(2) adrenergic receptor-mediated effect related to the release of nitric oxide. PMID- 17110425 TI - Differential effects of maternal dexamethasone treatment on circulating thyroid hormone concentrations and tissue deiodinase activity in the pregnant ewe and fetus. AB - Clinically, treatment of pregnant women at risk of preterm delivery with synthetic glucocorticoids accelerates fetal maturation. This study investigated the effect of maternal dexamethasone treatment, in clinically relevant doses, on plasma thyroid hormone concentrations and tissue deiodinase activities (D1, D2, and D3) in ewes and their fetuses. From 125 d of gestation (term 145 +/- 2 d), pregnant ewes were injected twice im with either saline (2 ml of 0.9% NaCl, n = 11) or dexamethasone (2 x 12 mg in 2 ml of saline, n = 10) at 24-h intervals. Maternal dexamethasone treatment increased plasma T(3) and reverse T(3) (rT(3)), but not T(4), concentrations in the fetuses. In the dexamethasone-exposed fetuses, hepatic D1 activity was higher, and renal and placental D3 activities were lower, than in the saline-exposed fetuses. In the ewes, plasma concentrations of T(3) and T(4) were reduced, and rT(3) increased, by dexamethasone treatment without any change in tissue deiodinase activity. Therefore, maternal dexamethasone treatment has different effects on the thyroid hormone axis of the pregnant ewe and fetus. In the fetus, the dexamethasone induced rise in circulating T(3) may be due to both increased hepatic production of T(3) from T(4), and reduced clearance of T(3) by the kidney and placenta. Changes in T(3) bioavailability may mediate some of the maturational effects of antenatal glucocorticoid treatment in the preterm fetus. PMID- 17110426 TI - Intestinal resistance to 1,25 dihydroxyvitamin D in mice heterozygous for the vitamin D receptor knockout allele. AB - We tested the hypothesis that low vitamin D receptor (VDR) level causes intestinal vitamin D resistance and intestinal calcium (Ca) malabsorption. To do so, we examined vitamin D regulated duodenal Ca absorption and gene expression [transient receptor potential channel, vallinoid subfamily member 6 (TRPV6), 24 hydroxylase, calbindin D(9k) (CaBP) mRNA, and CaBP protein] in wild-type mice and mice with reduced tissue VDR levels [i.e. heterozygotes for the VDR gene knockout (HT)]. Induction of 24-hydroxylase mRNA levels by 1,25 dihydroxyvitamin D(3) [1,25(OH)(2) D(3)] injection was significantly reduced in the duodenum and kidney of HT mice in both time-course and dose-response experiments. TRPV6 and CaBP mRNA levels in duodenum were significantly induced after 1,25(OH)(2) D(3) injection, but there was no difference in response between wild-type and HT mice. Feeding a low-calcium diet for 1 wk increased plasma PTH, renal 1alpha-hydroxylase (CYP27B1) mRNA level, and plasma 1,25(OH)(2) D(3), and this response was greater in HT mice (by 88, 55, and 37% higher, respectively). In contrast, duodenal TRPV6 and CaBP mRNA were not higher in HT mice fed the low-calcium diet. However, the response of duodenal Ca absorption and CaBP protein to increasing 1,25(OH)(2) D(3) levels was blunted by 40% in HT mice. Our data show that low VDR levels lead to resistance of intestinal Ca absorption to 1,25(OH)(2) D(3), and this resistance may be due to a role for the VDR (and VDR level) in the translation of CaBP. PMID- 17110427 TI - Adrenal insufficiency and colonic inflammation after a novel chronic psycho social stress paradigm in mice: implications and mechanisms. AB - We investigated chronic psycho-social stress effects on stress-related parameters and on pathohistological changes in the murine colon. Moreover, we aimed to reveal the involvement of adrenal glucocorticoids in chronic stress effects. Chronic subordinate colony housing (CSC, 19 d) resulted in reduced body weight gain, thymus atrophy, adrenal hypertrophy, increased plasma norepinephrine, and increased anxiety. With respect to the time course of CSC effects, CRH mRNA in the hypothalamic paraventricular nucleus, light phase corticosterone and tyrosine hydroxylase expression in colonic tissue were found to be increased, whereas tyrosine hydroxylase expression in the locus coeruleus was found to be decreased on d 2 of CSC; these parameters returned to control levels thereafter. Nevertheless, after 19 d of CSC exposure, the adrenal corticosterone responses in vivo and in vitro, and glucocorticoid sensitivity of isolated splenic cells were found to be decreased. Importantly, in CSC mice a significant histological damage of the colon was found beginning on d 14 of CSC exposure. Additionally, pro- and antiinflammatory cytokine secretion by mesenteric lymph node cells was increased after CSC exposure. Adrenalectomy before CSC at least partially prevented these chronic stress effects as reflected by less increase in proinflammatory cytokine secretion and an equal histological damage score in adrenalectomized compared with sham-operated CSC mice. In conclusion, chronic exposure to CSC alters relevant neuronal, neuroendocrine and immune functions that could be directly or indirectly involved in the damage of the histological integrity of the colon comparable with that seen during the development of colitis. PMID- 17110428 TI - Osteopontin protects the islets and beta-cells from interleukin-1 beta-mediated cytotoxicity through negative feedback regulation of nitric oxide. AB - Osteopontin (OPN), a phosphorylated glycoprotein that binds to an integrin binding motif, has been shown to regulate nitric oxide (NO) production via inhibition of induced NO synthase (iNOS) synthesis. In the transplanted islets, iNOS and toxic amounts of NO are produced as a result of islets infiltration with inflammatory cells and production of proinflammatory cytokines. Here, we demonstrate that addition of OPN before IL-1beta in freshly isolated rat islets improved their glucose stimulated insulin secretion dose-dependently and inhibited IL-1beta-induced NO production in an arginine-glycine-aspartate dependent manner. Transient transfection of OPN gene in RINm5F beta-cells fully prevented the toxic effect of IL-1beta at concentrations that reduced the viability by 50% over 3 d. OPN prevention of IL-1beta-induced toxicity was accompanied by inhibited transcription of iNOS by 80%, resulting in 50% decreased formation of the toxic NO. In OPN-transfected cells, the IL-1beta-induced nuclear factor-kappaB activity was significantly reduced. Islets exposed to IL-1beta revealed a naturally occurring early up-regulated OPN transcription. OPN promoter activity was increased in the presence of IL-1beta, IL-1beta-induced NO, and an inducer of NO synthesis. These data suggest the presence of a cross talk between the IL-1beta and OPN pathways and a unique trans-regulatory mechanism in which IL 1beta-induced NO synthesis feedback regulates itself through up-regulation of OPN gene transcription. Our data also suggest that influencing OPN expression represents an approach for affecting cytokine-induced signal transduction to prevent or reduce activation of the cascade of downstream devastating effects after islet transplantation. PMID- 17110429 TI - Follicle-stimulating hormone activates fatty acid amide hydrolase by protein kinase A and aromatase-dependent pathways in mouse primary Sertoli cells. AB - Among the biological activities of the endocannabinoid anandamide (N arachidonoylethanolamine) (AEA), growing interest has been attracted by the regulation of mammalian fertility. Recently we have shown that treatment of mouse primary Sertoli cells with FSH enhances the activity of the AEA hydrolase [fatty acid amide hydrolase (FAAH)], though the molecular details were not elucidated. Here, we investigated whether FSH was also able to affect the enzymes that synthesize AEA (N-acyltransferase and N-acyl-phosphatidyl-ethanolamine phospholipase D), the endogenous content of this endocannabinoid, and the level of the AEA-binding vanilloid receptor 1 (transient receptor potential channel vanilloid receptor subunit 1). We show that FSH enhanced FAAH activity (up to approximately 500% of the controls) and expression (up to approximately 300%), leading to a marked reduction (down to approximately 15%) of AEA content. However N-acyltransferase and N-acyl-phosphatidyl-ethanolamine-phospholipase D activity, and transient receptor potential channel vanilloid receptor subunit 1 binding were not affected. We also show that diacylglycerol lipase and monoacylglycerol lipase, which respectively synthesize and degrade 2-arachidonoyl-glycerol, were not regulated by FSH, neither was the membrane transport of this endocannabinoid. In addition, we show that FAAH stimulation by FSH was abrogated by inhibitors of protein kinase A (PKA) and cytochrome-P(450) aromatase, and was conversely mimicked by N,O'-dibutyryl cAMP and estrogen. Finally, we demonstrate that FSH protects Sertoli cells against the pro-apoptotic activity of AEA, through PKA and aromatase-dependent activation of FAAH. Altogether these data suggest that FAAH is the only target of FSH among the elements of the endocannabinoid system, and that its regulation by PKA and aromatase-dependent pathways impacts Sertoli cell proliferation. PMID- 17110430 TI - Identification of novel genes regulated by chorionic gonadotropin in baboon endometrium during the window of implantation. AB - Chorionic gonadotropin (CG) is an early embryo-derived signal that is known to support the corpus luteum. An in vivo baboon model was used to study the direct actions of human CG (hCG) on the endometrium, during the periimplantation period. Endometrial gene expression was analyzed using microarrays. The endometrial biopsies were taken from hCG-treated (n = 5) and control (n = 6) animals on d 10 after ovulation. Class comparison identified 61 genes whose transcript levels differed between control and hCG-treated samples (48 increased, 13 decreased in mean expression level more than 2.5-fold; P < 0.01). Real-time PCR of transcript abundance confirmed up-regulation of several of these, including SerpinA3, matrix metalloproteinase 7, leukemia inhibitory factor (LIF), IL-6, and Complement 3 (P 1,000 times more potent in generating anti-Pfs25H ELISA reactivity than a similar 0.5-microg dose of Pfs25H alone in Montanide ISA720, a water-in-oil adjuvant. The immune enhancement requires covalent conjugation between Pfs25H and the OMPC, given that physically mixed Pfs25H and OMPC on aluminum hydroxyphosphate failed to induce greater activity than the nonconjugated Pfs25H on aluminum hydroxyphosphate. The conjugate vaccine Pfs25H-OMPC also was highly immunogenic in rabbits and rhesus monkeys. In rhesus monkeys, the antibody responses were sustained over 18 months, at which time another vaccination with nonconjugated Pfs25H induced strong anamnestic responses. The vaccine-induced anti-Pfs25-specific antibodies in all animal species blocked the transmission of parasites to mosquitoes. Protein antigen conjugation to OMPC or other protein carrier may have general application to a spectrum of protein subunit vaccines to increase immunogenicity without the need for potentially reactogenic adjuvants. PMID- 17110441 TI - Munc18-1 expression levels control synapse recovery by regulating readily releasable pool size. AB - Prompt recovery after intense activity is an essential feature of most mammalian synapses. Here we show that synapses with reduced expression of the presynaptic gene munc18-1 suffer from increased depression during intense stimulation at glutamatergic, GABAergic, and neuromuscular synapses. Conversely, munc18-1 overexpression makes these synapses recover faster. Concomitant changes in the readily releasable vesicle pool and its refill kinetics were found. The number of vesicles docked at the active zone and the total number of vesicles per terminal correlated with both munc18-1 expression levels and the size of the releasable vesicle pool. These data show that varying expression of a single gene controls synaptic recovery by modulating the number of docked, release-ready vesicles and thereby replenishment of the secretion capacity. PMID- 17110442 TI - Ribosomal tethering and clustering as mechanisms for translation initiation. AB - Eukaryotic mRNAs often recruit ribosomal subunits some distance upstream of the initiation codon; however, the mechanisms by which they reach the initiation codon remain to be fully elucidated. Although scanning is a widely accepted model, evidence for alternative mechanisms has accumulated. We previously suggested that this process may involve tethering of ribosomal complexes to the mRNA, in which the intervening mRNA is bypassed, or clustering, in which the initiation codon is reached by dynamic binding and release of ribosomal subunits at internal sites. The present studies tested the feasibility of these ideas by using model mRNAs and revealed that translation efficiency varied with the distance between the site of ribosomal recruitment and the initiation codon. The present studies also showed that translation could initiate efficiently at AUG codons located upstream of an internal site. These observations are consistent with ribosomal tethering at the cap structure and clustering at internal sites. PMID- 17110443 TI - Alanine-glyoxylate aminotransferase-deficient mice, a model for primary hyperoxaluria that responds to adenoviral gene transfer. AB - Mutations in the alanine-glyoxylate amino transferase gene (AGXT) are responsible for primary hyperoxaluria type I, a rare disease characterized by excessive hepatic oxalate production that leads to renal failure. We generated a null mutant mouse by targeted mutagenesis of the homologous gene, Agxt, in embryonic stem cells. Mutant mice developed normally, and they exhibited hyperoxaluria and crystalluria. Approximately half of the male mice in mixed genetic background developed calcium oxalate urinary stones. Severe nephrocalcinosis and renal failure developed after enhancement of oxalate production by ethylene glycol administration. Hepatic expression of human AGT1, the protein encoded by AGXT, by adenoviral vector-mediated gene transfer in Agxt(-/-) mice normalized urinary oxalate excretion and prevented oxalate crystalluria. Subcellular fractionation and immunofluorescence studies revealed that, as in the human liver, the expressed wild-type human AGT1 was predominantly localized in mouse hepatocellular peroxisomes, whereas the most common mutant form of AGT1 (G170R) was localized predominantly in the mitochondria. PMID- 17110444 TI - Fragile X mental retardation protein controls trailer hitch expression and cleavage furrow formation in Drosophila embryos. AB - During the cleavage stage of animal embryogenesis, cell numbers increase dramatically without growth, and a shift from maternal to zygotic genetic control occurs called the midblastula transition. Although these processes are fundamental to animal development, the molecular mechanisms controlling them are poorly understood. Here, we demonstrate that Drosophila fragile X mental retardation protein (dFMRP) is required for cleavage furrow formation and functions within dynamic cytoplasmic ribonucleoprotein (RNP) bodies during the midblastula transition. dFMRP is observed to colocalize with the cytoplasmic RNP body components Maternal expression at 31B (ME31B) and Trailer Hitch (TRAL) in a punctate pattern throughout the cytoplasm of cleavage-stage embryos. Complementary biochemistry demonstrates that dFMRP does not associate with polyribosomes, consistent with their reported exclusion from many cytoplasmic RNP bodies. By using a conditional mutation in small bristles (sbr), which encodes an mRNA nuclear export factor, to disrupt the normal cytoplasmic accumulation of zygotic transcripts at the midblastula transition, we observe the formation of giant dFMRP/TRAL-associated structures, suggesting that dFMRP and TRAL dynamically regulate RNA metabolism at the midblastula transition. Furthermore, we show that dFMRP associates with endogenous tral mRNA and is required for normal TRAL protein expression and localization, revealing it as a previously undescribed target of dFMRP control. We also show genetically that tral itself is required for cleavage furrow formation. Together, these data suggest that in cleavage-stage Drosophila embryos, dFMRP affects protein expression by controlling the availability and/or competency of specific transcripts to be translated. PMID- 17110445 TI - Engineering cottonseed for use in human nutrition by tissue-specific reduction of toxic gossypol. AB - Global cottonseed production can potentially provide the protein requirements for half a billion people per year; however, it is woefully underutilized because of the presence of toxic gossypol within seed glands. Therefore, elimination of gossypol from cottonseed has been a long-standing goal of geneticists. Attempts were made to meet this objective by developing so-called "glandless cotton" in the 1950s by conventional breeding techniques; however, the glandless varieties were commercially unviable because of the increased susceptibility of the plant to insect pests due to the systemic absence of glands that contain gossypol and other protective terpenoids. Thus, the promise of cottonseed in contributing to the food requirements of the burgeoning world population remained unfulfilled. We have successfully used RNAi to disrupt gossypol biosynthesis in cottonseed tissue by interfering with the expression of the delta-cadinene synthase gene during seed development. We demonstrate that it is possible to significantly reduce cottonseed-gossypol levels in a stable and heritable manner. Results from enzyme activity and molecular analyses on developing transgenic embryos were consistent with the observed phenotype in the mature seeds. Most relevant, the levels of gossypol and related terpenoids in the foliage and floral parts were not diminished, and thus their potential function in plant defense against insects and diseases remained untouched. These results illustrate that a targeted genetic modification, applied to an underutilized agricultural byproduct, provides a mechanism to open up a new source of nutrition for hundreds of millions of people. PMID- 17110446 TI - Evidence for different origin of sex chromosomes in snakes, birds, and mammals and step-wise differentiation of snake sex chromosomes. AB - All snake species exhibit genetic sex determination with the ZZ/ZW type of sex chromosomes. To investigate the origin and evolution of snake sex chromosomes, we constructed, by FISH, a cytogenetic map of the Japanese four-striped rat snake (Elaphe quadrivirgata) with 109 cDNA clones. Eleven of the 109 clones were localized to the Z chromosome. All human and chicken homologues of the snake Z linked genes were located on autosomes, suggesting that the sex chromosomes of snakes, mammals, and birds were all derived from different autosomal pairs of the common ancestor. We mapped the 11 Z-linked genes of E. quadrivirgata to chromosomes of two other species, the Burmese python (Python molurus bivittatus) and the habu (Trimeresurus flavoviridis), to investigate the process of W chromosome differentiation. All and 3 of the 11 clones were localized to both the Z and W chromosomes in P. molurus and E. quadrivirgata, respectively, whereas no cDNA clones were mapped to the W chromosome in T. flavoviridis. Comparative mapping revealed that the sex chromosomes are only slightly differentiated in P. molurus, whereas they are fully differentiated in T. flavoviridis, and E. quadrivirgata is at a transitional stage of sex-chromosome differentiation. The differentiation of sex chromosomes was probably initiated from the distal region on the short arm of the protosex chromosome of the common ancestor, and then deletion and heterochromatization progressed on the sex-specific chromosome from the phylogenetically primitive boids to the more advanced viperids. PMID- 17110447 TI - Bioluminescence imaging of Smad signaling in living mice shows correlation with excitotoxic neurodegeneration. AB - The TGF-beta signaling pathway is a key organizer of injury and immune responses, and recent studies suggest it fulfills critical roles in CNS function and maintenance. TGF-beta receptor activation results in phosphorylation of Smad proteins, which subsequently translocate to the nucleus to regulate gene transcription by binding to Smad binding elements (SBE). Using SBE-luciferase reporter mice, we recently discovered that the brain has the highest Smad baseline activity of any major organ in the mouse, and we now demonstrate that this signal is primarily localized to pyramidal neurons of the hippocampus. In vivo excitatory stimulation with kainic acid (KA) resulted in an increase in luciferase activity and phosphorylated Smad2 (Smad2P), and nuclear translocation of Smad2P in hippocampal CA3 neurons correlated significantly with luciferase activity. Although this activation was most prominent at 24 h after KA administration in neurons, Smad2P immunoreactivity gradually increased in astrocytes and microglial cells at 3 and 5 days, consistent with reactive gliosis. Bioluminescence measured over the skull in living mice peaked at 12-72 h and correlated with the extent of microglial activation and pathological markers of neurodegeneration 5 days after injury. Treatment with the glutamate receptor antagonist MK-801 strongly reduced bioluminescence and pathology. These results show that Smad2 signaling is a sensitive marker of neuronal activation and CNS injury that can be used to monitor KA-induced neuronal degeneration. This and related mouse models may provide valuable tools to study mechanisms and treatments for neurodegeneration. PMID- 17110448 TI - The use of transcriptional profiles to predict adult mosquito age under field conditions. AB - Age is a critical determinant of an adult female mosquito's ability to transmit a range of human pathogens. Despite its central importance, relatively few methods exist with which to accurately determine chronological age of field-caught mosquitoes. This fact is a major constraint on our ability to fully understand the relative importance of vector longevity to disease transmission in different ecological contexts. It also limits our ability to evaluate novel disease control strategies that specifically target mosquito longevity. We report the development of a transcriptional profiling approach to determine age of adult female Aedes aegypti under field conditions. We demonstrate that this approach surpasses current cuticular hydrocarbon methods for both accuracy of predicted age as well as the upper limits at which age can be reliably predicted. The method is based on genes that display age-dependent expression in a range of dipteran insects and, as such, is likely to be broadly applicable to other disease vectors. PMID- 17110449 TI - Celastrol, a novel triterpene, potentiates TNF-induced apoptosis and suppresses invasion of tumor cells by inhibiting NF-kappaB-regulated gene products and TAK1 mediated NF-kappaB activation. AB - Celastrol, a quinone methide triterpene derived from the medicinal plant Tripterygium wilfordii, has been used to treat chronic inflammatory and autoimmune diseases, but its mechanism is not well understood. Therefore, we investigated the effects of celastrol on cellular responses activated by TNF, a potent proinflammatory cytokine. Celastrol potentiated the apoptosis induced by TNF and chemotherapeutic agents and inhibited invasion, both regulated by NF kappaB activation. We found that TNF induced the expression of gene products involved in antiapoptosis (IAP1, IAP2, Bcl-2, Bcl-XL, c-FLIP, and survivin), proliferation (cyclin D1 and COX-2), invasion (MMP-9), and angiogenesis (VEGF) and that celastrol treatment suppressed their expression. Because these gene products are regulated by NF-kappaB, we postulated that celastrol mediates its effects by modulating the NF-kappaB pathway. We found that celastrol suppressed both inducible and constitutive NF-kappaB activation. Celastrol was found to inhibit the TNF-induced activation of IkappaBalpha kinase, IkappaBalpha phosphorylation, IkappaBalpha degradation, p65 nuclear translocation and phosphorylation, and NF-kappaB-mediated reporter gene expression. Recent studies indicate that TNF-induced IKK activation requires activation of TAK1, and we indeed found that celastrol inhibited the TAK1-induced NF-kappaB activation. Overall, our results suggest that celastrol potentiates TNF-induced apoptosis and inhibits invasion through suppression of the NF-kappaB pathway. PMID- 17110450 TI - Hlx homeobox transcription factor negatively regulates interferon-gamma production in monokine-activated natural killer cells. AB - Natural killer (NK) cells contribute to host immunity, including tumor surveillance, through the production of interferon gamma (IFN-gamma). Although there is some knowledge about molecular mechanisms that induce IFN-gamma in NK cells, considerably less is known about the mechanisms that reduce its expression. Here, we investigate the role of the Hlx transcription factor in IFN gamma production by NK cells. Hlx expression is induced in monokine-activated NK cells, but with delayed kinetics compared to IFN-gamma. Ectopic Hlx expression decreases IFN-gamma synthesis in primary human NK cells and IFN-gamma promoter activity in an NK-like cell line. Hlx protein levels inversely correlate with those of STAT4, a requisite factor for optimal IFN-gamma transcription. Mechanistically, we provide evidence indicating that Hlx overexpression accelerates dephosphorylation and proteasome-dependent degradation of the active Y693-phosphorylated form of STAT4. Thus, Hlx expression in activated NK cells temporally controls and limits the monokine-induced production of IFN-gamma, in part through the targeted depletion of STAT4. PMID- 17110451 TI - Vitamin K supplementation can improve stability of anticoagulation for patients with unexplained variability in response to warfarin. AB - Patients receiving warfarin who have unstable control of anticoagulation have a significantly lower intake of dietary vitamin K compared with their stable counterparts. We hypothesized that supplementation with oral vitamin K would improve stability in patients with previously unstable control of anticoagulation. Seventy warfarin-treated patients with unstable anticoagulation control were randomly assigned in a double-blinded fashion to receive a daily amount of 150 mug oral vitamin K or placebo orally for 6 months. Measures of stability of anticoagulation control in the 6-month study period were compared with those in the 6 months immediately prior to it. Vitamin K supplementation resulted in a significantly greater decrease in standard deviation of international normalized ratio (INR) compared with placebo (-0.24 +/- 0.14 vs 0.11 +/- 0.18; P < .001) and a significantly greater increase in percentage time within target INR range (28% +/- 20% vs 15% +/- 20%; P < .01). Anticoagulation control improved in 33 of 35 patients receiving vitamin K supplementation; of these, 19 fulfilled our criteria for having stable control of anticoagulation. However, only 24 of 33 patients receiving placebo demonstrated some degree of improvement, with only 7 patients fulfilling the criteria for having stable control. Concomitant supplementation of vitamin K, perhaps through reducing the relative day-to-day variability in dietary vitamin K intake, can significantly improve anticoagulation control in patients with unexplained instability of response to warfarin. PMID- 17110452 TI - Leukocytosis is a risk factor for thrombosis in essential thrombocythemia: interaction with treatment, standard risk factors, and Jak2 mutation status. AB - Leukocytes contribute to the pathogenesis of thrombosis in essential thrombocythemia (ET) through recently discovered mechanisms of activation and interaction with platelets and endothelial cells. To evaluate whether an increased leukocyte count was associated with thrombosis and whether this effect can be modulated by therapy, we analyzed the clinical course of 439 patients with ET followed at the Ospedali Riuniti di Bergamo. The strength of the association was measured at diagnosis or before thrombotic events by multivariable analyses carried out using data at baseline as well as time-varying covariates. The results showed that (1) an increased leukocyte count at diagnosis was associated with thrombosis during follow-up ("baseline analysis," relative risk [RR] 2.3, 95% confidence interval [CI] 1.4-3.9, P = .001); (2) hydroxyurea (HU) lowered leukocytosis and reduced the strength of the association between leukocytosis and thrombosis ("time-dependent analysis," RR 1.6, 95% CI 0.9-2.0, not significant [NS]); (3) the association of leukocytosis and thrombosis was more evident in untreated low-risk patients (RR 2.7, 95% CI 1.2-6.4, P = .01) compared with HU treated high-risk patients (RR 1.6, 95% CI 0.8-3.2, NS); and (4) the presence of JAK2 V617F was not identified as a risk factor for thrombosis during follow-up despite a significant association between the mutation and leukocytosis. We suggest validation of these findings in prospective clinical studies. PMID- 17110453 TI - The cytoprotective protein C pathway. AB - Protein C is best known for its mild deficiency associated with venous thrombosis risk and severe deficiency associated with neonatal purpura fulminans. Activated protein C (APC) anticoagulant activity involves proteolytic inactivation of factors Va and VIIIa, and APC resistance is often caused by factor V Leiden. Less known is the clinical success of APC in reducing mortality in severe sepsis patients (PROWESS trial) that gave impetus to new directions for basic and preclinical research on APC. This review summarizes insights gleaned from recent in vitro and in vivo studies of the direct cytoprotective effects of APC that include beneficial alterations in gene expression profiles, anti-inflammatory actions, antiapoptotic activities, and stabilization of endothelial barriers. APC's cytoprotection requires its receptor, endothelial cell protein C receptor, and protease-activated receptor-1. Because of its pleiotropic activities, APC has potential roles in the treatment of complex disorders, including sepsis, thrombosis, and ischemic stroke. Although much about molecular mechanisms for APC's effects on cells remains unclear, it is clear that APC's structural features mediating anticoagulant actions and related bleeding risks are distinct from those mediating cytoprotective actions, suggesting the possibility of developing APC variants with an improved profile for the ratio of cytoprotective to anticoagulant actions. PMID- 17110454 TI - mTOR-dependent synthesis of Bcl-3 controls the retraction of fibrin clots by activated human platelets. AB - New activities of human platelets continue to emerge. One unexpected response is new synthesis of proteins from previously transcribed RNAs in response to activating signals. We previously reported that activated human platelets synthesize B-cell lymphoma-3 (Bcl-3) under translational control by mammalian target of rapamycin (mTOR). Characterization of the ontogeny and distribution of the mTOR signaling pathway in CD34+ stem cell-derived megakaryocytes now demonstrates that they transfer this regulatory system to developing proplatelets. We also found that Bcl-3 is required for condensation of fibrin by activated platelets, demonstrating functional significance for mTOR-regulated synthesis of the protein. Inhibition of mTOR by rapamycin blocks clot retraction by human platelets. Platelets from wild-type mice synthesize Bcl-3 in response to activation, as do human platelets, and platelets from mice with targeted deletion of Bcl-3 have defective retraction of fibrin in platelet-fibrin clots mimicking treatment of human platelets with rapamycin. In contrast, overexpression of Bcl-3 in a surrogate cell line enhanced clot retraction. These studies identify new features of post-transcriptional gene regulation and signal-dependant protein synthesis in activated platelets that may contribute to thrombus and wound remodeling and suggest that posttranscriptional pathways are targets for molecular intervention in thrombotic disorders. PMID- 17110455 TI - A coding VKORC1 Asp36Tyr polymorphism predisposes to warfarin resistance. AB - CYP2C9 and VKORC1 genetic variants are associated with low and intermediate warfarin dose requirements, but markers of high doses are less well characterized. We analyzed the VKORC1 coding sequence and known CYP2C9 and VKORC1 polymorphisms in 15 selected warfarin-resistant (dose, 80 to 185 mg/wk) and 8 warfarin-sensitive patients (7 to 13 mg/wk) and 99 unselected controls (8 to 105 mg/wk). We identified a coding VKORC1 Asp36Tyr polymorphism in 7 of 15 resistant compared with 0 of 8 sensitive patients (P = .026) Carriers of Asp36Tyr in the control group (8 of 99) required significantly higher warfarin doses of 80.9 +/- 10.1 mg/wk compared with 42.7 +/- 7.5 mg/wk in noncarriers (F = 9.79, P = .002). Asp36Tyr was significantly associated with doses of more than 70 mg/wk (odds ratio, 13.0; 95% confidence limit, 1.3 to 124.2), while doses of 20 to 70 mg/wk were associated with Asp36Tyr (partial r(2) = .11; P = .004), CYP2C9*2 and *3 (r(2) = .08; P = .01), and VKORC1*2 and *3 markers (r(2) = .05; P = .05). All Asp36Tyr carriers also had VKORC1*1 tag-single nucleotide polymorphisms (tag SNPs) indicating a new haplotype. Asp36Tyr was common in Jewish ethnic groups of Ethiopian (15%) and Ashkenazi (4%) origin. We suggest that Asp36Tyr is a new marker of the high end of the warfarin dosing range. PMID- 17110457 TI - A phase 2/3 multicenter randomized clinical trial of ABX-CBL versus ATG as secondary therapy for steroid-resistant acute graft-versus-host disease. AB - Treatment for steroid-resistant acute graft-versus-host disease (GVHD) has had limited success. ABX-CBL is a hybridoma-generated murine IgM monoclonal antibody against the CD147 antigen, weakly expressed on human leukocytes and up-regulated on activated lymphocytes. A prospective, multicenter, open-label, randomized clinical trial comparing ABX-CBL to antithymocyte globulin (ATG) for treatment of steroid-resistant acute GVHD was conducted in 95 patients at 21 centers. Forty eight patients received ABX-CBL daily for 14 consecutive days followed by up to 6 weeks of ABX-CBL twice weekly. Forty-seven patients received equine ATG, 30 mg/kg every other day for a total of 6 doses with additional courses as needed. By day 180, overall improvement was similar in the patients receiving ABX-CBL and in those receiving ATG (56% versus 57%, P = .91). Patient survival at 18 months was less favorable on ABX-CBL than on ATG (35% versus 45%), with the 95% confidence interval ruling out that ABX-CBL provides at least a 10.4% improvement. Data from this trial suggest that ABX-CBL does not offer an improvement over ATG in the treatment of acute steroid-resistant GVHD. This prospective, multicenter, randomized clinical trial for steroid-resistant acute GVHD serves as a model for future evaluation of new agents. PMID- 17110456 TI - Regulation of protein synthesis by the heme-regulated eIF2alpha kinase: relevance to anemias. AB - During erythroid differentiation and maturation, it is critical that the 3 components of hemoglobin, alpha-globin, beta-globin, and heme, are made in proper stoichiometry to form stable hemoglobin. Heme-regulated translation mediated by the heme-regulated inhibitor kinase (HRI) provides one major mechanism that ensures balanced synthesis of globins and heme. HRI phosphorylates the alpha subunit of eukaryotic translational initiation factor 2 (eLF2alpha) in heme deficiency, thereby inhibiting protein synthesis globally. In this manner, HRI serves as a feedback inhibitor of globin synthesis by sensing the intracellular concentration of heme through its heme-binding domains. HRI is essential not only for the translational regulation of globins, but also for the survival of erythroid precursors in iron deficiency. Recently, the protective function of HRI has also been demonstrated in murine models of erythropoietic protoporphyria and beta-thalassemia. In these 3 anemias, HRI is essential in determining red blood cell size, number, and hemoglobin content per cell. Translational regulation by HRI is critical to reduce excess synthesis of globin proteins or heme under nonoptimal disease states, and thus reduces the severity of these diseases. The protective role of HRI may be more common among red cell disorders. PMID- 17110458 TI - Specific antibodies to moesin, a membrane-cytoskeleton linker protein, are frequently detected in patients with acquired aplastic anemia. AB - To identify novel autoantibodies in acquired aplastic anemia (AA), we screened the sera of patients with AA possessing small populations of paroxysmal nocturnal hemoglobinuria (PNH)-type cells for the presence of antibodies (Abs) which recognize proteins derived from a leukemia cell line, UT-7. Immunoblotting using proteins derived from lysates or culture supernatants of UT-7 cells revealed the presence of IgG Abs specific to an 80-kDa protein. Peptide mass fingerprinting identified this 80-kDa protein as moesin. Enzyme-linked immunosorbent assay (ELISA) using recombinant moesin showed high titers of antimoesin Abs in 25 (37%) of 67 patients with AA. Moesin was secreted from several myeloid leukemia cell lines other than UT-7, such as OUN-1 and K562, as an exosomal protein. The presence of antimoesin Abs was significantly correlated with the presence of PNH type cells and antidiazepam-binding inhibitor-related protein-1 (DRS-1) Abs. Patients with AA that did not show any of these 3 markers tended to respond poorly to immunosuppressive therapy. These findings suggest that a B-cell response to moesin, possibly derived from hematopoietic cells, frequently occurs in patients with AA and that detection of antimoesin Abs in combination with other markers may be useful in diagnosing immune pathophysiology in patients with AA. PMID- 17110459 TI - Granulocyte-stimulating factor and severe aplastic anemia: a survey by the European Group for Blood and Marrow Transplantation (EBMT). AB - Previous studies suggested a link between the use of G-CSF and increased incidence of myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) after immunosuppressive therapy (IST) for severe aplastic anemia (SAA). This European survey included 840 patients who received a first-line IST with (43%) or without (57%) G-CSF. The incidences of MDS/AML in patients who did or did not receive G-CSF were 10.9% and 5.8%, respectively. A significantly higher hazard (1.9) of MDS/AML was associated with use of G-CSF. Relapse of aplastic anemia was not associated with a worse outcome in patients who did not receive G-CSF as first therapy, whereas relapse was associated with a significantly worse outcome in those patients who received G-CSF. These results emphasize the necessity of the current European randomized trial comparing IST with or without G-CSF and to alert physicians that adding G-CSF to IST is currently not standard treatment for SAA. PMID- 17110460 TI - Identification of MCL1 as a novel target in neoplastic mast cells in systemic mastocytosis: inhibition of mast cell survival by MCL1 antisense oligonucleotides and synergism with PKC412. AB - MCL-1 is a Bcl-2 family member that has been described as antiapoptotic in various myeloid neoplasms. Therefore, MCL-1 has been suggested as a potential new therapeutic target. Systemic mastocytosis (SM) is a myeloid neoplasm involving mast cells (MCs) and their progenitors. In the present study, we examined the expression and functional role of MCL-1 in neoplastic MCs and sought to determine whether MCL-1 could serve as a target in SM. As assessed by RT-PCR and immunohistochemical examination, primary neoplastic MCs expressed MCL-1 mRNA and the MCL-1 protein in all SM patients examined. Moreover, MCL-1 was detectable in both subclones of the MC line HMC-1--HMC-1.1 cells, which lack the SM-related KIT mutation D816V, and HMC-1.2 cells, which carry KIT D816V. Exposure of HMC-1.1 cells or HMC-1.2 cells to MCL-1-specific antisense oligonucleotides (ASOs) or MCL 1-specific siRNA resulted in reduced survival and increased apoptosis compared with untreated cells. Moreover, MCL-1 ASOs were found to cooperate with various tyrosine kinase inhibitors in producing growth inhibition in neoplastic MCs, with synergistic effects observed with PKC412, AMN107, and imatinib in HMC-1.1 cells and with PKC412 in HMC-1.2 cells. Together, these data show that MCL-1 is a novel survival factor and an attractive target in neoplastic MCs. PMID- 17110461 TI - Hypoxia-inducible transcription factor-1 alpha determines sensitivity of endothelial cells to the proteosome inhibitor bortezomib. AB - Angiogenesis is a complex, orchestrated process that plays a critical role in several conditions and has special relevance in the progression of cancer. Hypoxia is the major stimulus for angiogenesis, and hypoxia-inducible transcription factor-1 alpha (HIF-1alpha) is its key mediator. We set up a novel in vitro model of HIF-1alpha up-regulation by treating human umbilical vein endothelial cells (HUVECs) with the hypoxia-mimicking deferoxamine (DFO) and found that this condition was sufficient to promote angiogenesis, like the well known HUVEC model cultured under low pO(2.) The proteasome inhibitor bortezomib, which induces strong apoptosis in cancer cells, abrogated proliferation and angiogenesis of HUVECs when used at a high concentration (100 nM), yet promoted both functions at a low dosage (10 nM). This double-edged effect appeared to be mediated by differential effects exerted by the different concentrations of bortezomib on 2 master regulators of tumor-associated angiogenesis, HIF-1alpha and nuclear factor kappa B (NF-kB). Significantly, when HUVECs were induced to express HIF-1alpha prior to bortezomib treatment, proliferative and angiogenic responses were abolished, and a greatly enhanced proapoptotic effect was promoted with both concentrations of the drug. These findings indicate that HIF-1alpha up regulation may sensitize endothelial cells to the antiangiogenic and proapoptotic effects of bortezomib and might be exploited to target tumor-associated vessels in the course of antiangiogenic therapies. PMID- 17110462 TI - Galectin-1: a key effector of regulation mediated by CD4+CD25+ T cells. AB - The naturally occurring population of dedicated regulatory T cells that coexpress CD4 and CD25 is known to play a key role in the maintenance of peripheral T-cell tolerance; however, their mechanism of action has remained obscure. Here we report that a member of the family of beta-galactoside-binding proteins, galectin 1, is overexpressed in regulatory T cells, and that expression is increased after activation. Most importantly, blockade of galectin-1 binding significantly reduced the inhibitory effects of human and mouse CD4+CD25+ T cells. Reduced regulatory activity was observed in CD4+CD25+ T cells obtained from galectin-1 homozygous null mutant mice. These results suggest that galectin-1 is a key effector of the regulation mediated by these cells. PMID- 17110463 TI - Expression of a non-DNA-binding isoform of Helios induces T-cell lymphoma in mice. AB - Helios is a zinc-finger protein belonging to the Ikaros family of transcriptional regulators. It is expressed, along with Ikaros, throughout early stages of thymocyte development where it quantitatively associates with Ikaros through C terminal zinc-finger domains that mediate heterodimerization between Ikaros family members. To understand the role of Helios in T-cell development, we used a retroviral vector to express full-length Helios or a Helios isoform that lacked the N-terminal DNA-binding domain in hematopoietic progenitor cells of reconstituted mice. Constitutive expression of full-length Helios resulted in an inhibition of T-cell development at the double-negative stage within the thymus. Although expression of the DNA-binding mutant of Helios did not contribute to developmental abnormalities at early times after transplantation, 60% of animals that expressed the Helios DNA-binding mutant developed an aggressive and transplantable T-cell lymphoma 4 to 10 months after transplantation. These results demonstrate a vital function for Helios in maintaining normal homeostasis of developing T cells and formally show that non-DNA-binding isoforms of Helios are lymphomagenic if aberrantly expressed within the T-cell lineage. PMID- 17110464 TI - A combined empirical and mechanistic codon model. AB - The evolutionary selection forces acting on a protein are commonly inferred using evolutionary codon models by contrasting the rate of synonymous to nonsynonymous substitutions. Most widely used models are based on theoretical assumptions and ignore the empirical observation that distinct amino acids differ in their replacement rates. In this paper, we develop a general method that allows assimilation of empirical amino acid replacement probabilities into a codon substitution matrix. In this way, the resulting codon model takes into account not only the transition-transversion bias and the nonsynonymous/synonymous ratio, but also the different amino acid replacement probabilities as specified in empirical amino acid matrices. Different empirical amino acid replacement matrices, such as secondary structure-specific matrices or organelle-specific matrices (e.g., mitochondria and chloroplasts), can be incorporated into the model, making it context dependent. Using a diverse set of coding DNA sequences, we show that the novel model better fits biological data as compared with either mechanistic or empirical codon models. Using the suggested model, we further analyze human immunodeficiency virus type 1 protease sequences obtained from drug treated patients and reveal positive selection in sites that are known to confer drug resistance to the virus. PMID- 17110465 TI - Defining a binding pocket for sulfonylureas in ATP-sensitive potassium channels. AB - Sulfonylurea receptors SUR1 and SUR2 are the regulatory subunits of K(ATP) channels. Their differential affinity for hypoglycemic sulfonylureas provides a basis for the selectivity of these compounds for different K(ATP) channel isoforms. Sulfonylureas have a 100- to 1000-fold greater affinity for SUR1 vs. SUR2. Structure-activity studies suggested a bipartite binding pocket. Chimeric SUR1 approximately SUR2 receptors have shown TMD2, the third bundle of transmembrane helices, to be part of an "A" site that confers SUR1 selectivity for sulfonylureas. The purpose of this study is to determine the position of the "B" site. Previous photoaffinity labeling studies have placed the B site on the amino-terminal third of SUR and colabeled the associated K(IR). In our study, deletion of TMD0, the first bundle of transmembrane helices, did not compromise labeling. Further deletions into the cytoplasmic linker, L0, eliminated binding and labeling. Alanine substitutions in L0 identified a limited number of conserved residues, Y230 and W232, important for affinity labeling. A fragment of K(IR)6.2, missing M2 and the entire carboxyl terminal, assembles with SUR1 and is affinity labeled, while deletion of 10 or more amino-terminal residues compromises labeling. These studies indicate that the B site involves L0 and the K(IR) amino terminus, elements that are critical for control of channel gating. PMID- 17110466 TI - Dihydrolipoyl dehydrogenase as a source of reactive oxygen species inhibited by caloric restriction and involved in Saccharomyces cerevisiae aging. AB - Replicative life span in Saccharomyces cerevisiae is increased by glucose (Glc) limitation [calorie restriction (CR)] and by augmented NAD+. Increased survival promoted by CR was attributed previously to the NAD+-dependent histone deacetylase activity of sirtuin family protein Sir2p but not to changes in redox state. Here we show that strains defective in NAD+ synthesis and salvage pathways (pnc1delta, npt1delta, and bna6delta) exhibit decreased oxygen consumption and increased mitochondrial H2O2 release, reversed over time by CR. These null mutant strains also present decreased chronological longevity in a manner rescued by CR. Furthermore, we observed that changes in mitochondrial H2O2 release alter cellular redox state, as attested by measurements of total, oxidized, and reduced glutathione. Surprisingly, our results indicate that matrix-soluble dihydrolipoyl dehydrogenases are an important source of CR-preventable mitochondrial reactive oxygen species (ROS). Indeed, deletion of the LPD1 gene prevented oxidative stress in npt1delta and bna6delta mutants. Furthermore, pyruvate and alpha ketoglutarate, substrates for dihydrolipoyl dehydrogenase-containing enzymes, promoted pronounced reactive oxygen release in permeabilized wild-type mitochondria. Altogether, these results substantiate the concept that mitochondrial ROS can be limited by caloric restriction and play an important role in S. cerevisiae senescence. Furthermore, these findings uncover dihydrolipoyl dehydrogenase as an important and novel source of ROS leading to life span limitation. PMID- 17110467 TI - Effect of olive oils on biomarkers of oxidative DNA stress in Northern and Southern Europeans. AB - High consumption of olive oil in the Mediterranean diet has been suggested to protect DNA against oxidative damage and to reduce cancer incidence. We investigated the impact of the phenolic compounds in olive oil, and the oil proper, on DNA and RNA oxidation in North, Central, and South European populations. In a multicenter, double-blind, randomized, controlled crossover intervention trial, the effect of olive oil phenolic content on urinary oxidation products of guanine (8-oxo-guanine, 8-oxo-guanosine and 8-oxo-deoxyguanosine) was investigated. Twenty-five milliliters of three olive oils with low, medium, and high phenolic content were administered to healthy males (n=182) daily for 3 wk. At study baseline the urinary excretion of 8-oxo-guanosine (RNA oxidation) and 8 oxo-deoxyguanosine (DNA oxidation) was higher in the Northern regions of Europe compared with Central and Southern European regions (P=0.035). Urinary excretion of the 8 hydroxylated forms of guanine, guanosine, deoxyguanosine and their nonoxidized forms were not different when comparing olive oils with low, medium, and high phenolic content given for 2 wk. Testing the effect of oil from urinary 8-oxo-deoxyguanosine changes from baseline to post-treatment showed a reduction of DNA oxidation by 13% (P=0.008). These findings support the idea that ingestion of olive oil is beneficial and can reduce the rate of oxidation of DNA. This effect is not due to the phenolic content in the olive oil. The higher DNA and RNA oxidation in Northern European regions compared with that in Central and Southern regions supports the contention that olive oil consumption may explain some of the North-South differences in cancer incidences in Europe. PMID- 17110468 TI - Islet-derived fibroblast-like cells are not derived via epithelial-mesenchymal transition from Pdx-1 or insulin-positive cells. AB - As recent studies suggest that newly formed pancreatic beta-cells are a result of self-duplication rather than stem cell differentiation, in vitro expansion of beta-cells presents a potential mechanism by which to increase available donor tissue for cell-based diabetes therapies. Although most studies have found that beta-cells are resilient to substantial in vitro expansion, recent studies have suggested that it is possible to expand these cells through a process referred to as epithelial-mesenchymal transition (EMT). To further substantiate such an expansion mechanism, we used recombination-based genetic lineage tracing to determine the origin of proliferating fibroblast-like cells from cultured pancreatic islets in vitro. We demonstrate, using two culture methods, that EMT does not underlie the appearance of fibroblast-like cells in mouse islet cultures but that fibroblast-like cells appear to represent mesenchymal stem cell (MSC) like cells akin to MSCs isolated from bone marrow. PMID- 17110469 TI - Coexistence of (partial) immune defects and risk of recurrent respiratory infections. AB - BACKGROUND: Respiratory infections are major causes of morbidity and mortality, but determinants of susceptibility are poorly defined. We studied whether and to what extent immunologic and genetic factors are associated with increased susceptibility to respiratory infections. METHODS: We evaluated the prevalence of IgA, IgM, IgG, and IgG subclass deficiencies, impairment in the antibody response against pneumococcal polysaccharides, G2m(n) allotypes, Fc gamma RIIa polymorphisms, partial C2 and partial C4 deficiency, promoter polymorphisms in MBL2, and lymphocyte subset deficiencies in a control population and in consecutive children with recurrent respiratory infections. RESULTS: IgA and/or IgG subclass deficiency was found in 27 of 55 patients (49%) and 6 of 43 controls (14%) (P = 0.0006). An impaired antibody response to polysaccharides was found in 7 patients (19%) and in 0 of 37 controls (P = 0.002). The Gm(n)marker was absent in 25 of 55 patients (45%) and 6 of 42 controls (14%) (P = 0.009). The MBL2 variants O/O, A/O, and A/A occurred in 9, 14, and 32 of the 55 patients, respectively, and in 1, 19, and 23 of the 43 controls, respectively (P = 0.05). There was no increase in the prevalence of partial C4 deficiency, C2 deficiency, lymphocyte subset deficiency, or Fc gamma RIIa polymorphism in the patients compared to the controls. A combination of at least 2 immune defects was found in 31 of 55 patients (56%) and in 4 of 42 controls (11.6%) (P <0.0001). CONCLUSION: Specific antipolysaccharide antibody deficiency, IgA and/or IgG subclass deficiency, Gm(n) allotype, and MBL2 genotype are susceptibility factors for recurrent respiratory infections, and coexistence of several immune defects is the strongest risk factor in this study. PMID- 17110470 TI - gamma-Glutamyltransferase as a predictor of chronic kidney disease in nonhypertensive and nondiabetic Korean men. AB - BACKGROUND: Little research has been done to examine whether gamma glutamyltransferase (GGT) is prospectively associated with the development of chronic kidney disease (CKD). We performed a prospective study to examine the association between GGT and the risk for the development of CKD. METHODS: The study cohort included a total of 10 337 healthy males with normal baseline kidney functions and no proteinuria. Participants were workers in a semiconductor manufacturing company and its 13 affiliates. CKD was defined as either the presence of proteinuria or a glomerular filtration rate (GFR) of < 60 mL x min( 1) x (1.73(2))(-1). Cox proportional hazards models were used to calculate the adjusted hazard ratios in separate models for CKD. RESULTS: During a follow-up period of 25,774.4 person-years, 366 men developed CKD. After adjustments were made for age, baseline GFR, triglyceride, and HDL-C, the risk for CKD increased with an increasing quartile of serum GGT (p for trend <0.001). The top one fourth of serum GGT vs the bottom one fourth of relative risks for CKD was 1.90 (95% confidence interval, 1.37-2.63). These associations were also apparent in participants who consumed < or = 20 g/day of alcohol and those with normal weight, with values of alanine aminotransferase within reference intervals, or with C-reactive protein < 3.0 mg/L, and participants without metabolic syndrome. CONCLUSIONS: Our findings, which were obtained from a large work-site cohort and excluded individuals with diabetes and hypertension, indicated that serum GGT may be an early predictor for the development of CKD, independent of baseline confounding factors. PMID- 17110471 TI - Factitious diarrhea induced by stimulant laxatives: accuracy of diagnosis by a clinical reference laboratory using thin layer chromatography. AB - BACKGROUND: Surreptitious ingestion of laxatives can lead to serious factitious diseases that are difficult to diagnose. Most cases involve ingestion of bisacodyl or senna. Thin layer chromatography (TLC) of urine or stool is the only commercially available test for these laxatives. Such testing is considered highly reliable, but its accuracy in clinical practice is unknown. Our aim was to evaluate the reliability of TLC laxative testing by a clinical reference laboratory in the United States. METHODS: Diarrhea was induced in healthy volunteers by ingestion of bisacodyl, senna, or a control laxative (n = 11 for each laxative group). Samples of urine and diarrheal stool were sent in blinded fashion to the clinical reference laboratory for bisacodyl and senna analysis. RESULTS: TLC testing for bisacodyl-induced diarrhea revealed a sensitivity of 73% and specificity of 91% when urine was tested and sensitivity and specificity of 91% and 96%, respectively, when stool was analyzed. When diarrhea was induced by senna, the TLC assay for senna failed to identify even a single urine or stool specimen as positive (zero% sensitivity). CONCLUSIONS: Considering the expected prevalence of surreptitious laxative abuse in patients with chronic idiopathic diarrhea (2.4%-25%, depending on the clinical setting), TLC of urine or stool for bisacodyl by this reference laboratory would often produce misleading results, and testing for senna would have no clinical value. The major problems are false positive tests for bisacodyl and false-negative tests for senna. PMID- 17110472 TI - Molecular beacon-based temperature control and automated analyses for improved resolution of melting temperature analysis using SYBR I green chemistry. AB - BACKGROUND: Melting temperature analysis of products amplified with SYBR I Green chemistry is a cheap and effective method for identification of sequence differences. When used in conventional quantitative real-time PCR instruments (qPCR), this method is limited by temperature variations over the heating block and low numbers of fluorescence measurements during the dissociation step, which hamper the ability of most instruments to report accurate and precise melting temperatures. METHODS: We designed a molecular beacon-based temperature indicator probe (Tm-probe) to control for variations in temperatures over the heating block of the instrument. In addition, we wrote an automated curve-fit analysis algorithm of dissociation data to use multiple data points with a gaussian curve fit to extrapolate precise melting temperatures. RESULTS: Use of the Tm-probe in conjunction with the analysis algorithm and multiple dissociations improved SDs of melting temperatures over a 96-well plate from 0.19 to 0.06 degrees C. CONCLUSIONS: Melting temperature analyses with SYBR I Green chemistry on conventional qPCR instruments can be improved by the use of a Tm-probe in conjunction with curve-fit analysis of data. Resolution improvement up to 3-fold is possible and allows additional melting temperatures to be identified. PMID- 17110473 TI - Endothelial nitric oxide synthase haplotypes are associated with features of metabolic syndrome. AB - BACKGROUND: The metabolic syndrome, a cluster of several metabolic disorders, is increasingly being recognized as a risk factor for cardiovascular disease. Endothelium-derived nitric oxide facilitates skeletal muscle glucose uptake, and data from animal models indicate that endothelial nitric oxide synthase (eNOS) gene-null mice present with a phenotype of insulin resistance, hypertension, and hypertriglyceridemia, much like that observed in humans with metabolic syndrome. We used haplotype tagging single nucleotide polymorphisms (htSNPs) to investigate the role of genetic variation in the eNOS gene (NOS3) in metabolic syndrome in humans. METHODS: We recruited 738 unrelated persons from a cross-sectional population-based epidemiological survey in the province of Segovia in Central Spain (Castille). Metabolic syndrome was defined according to the recently modified National Cholesterol Education Program Adult Treatment Panel III guidelines. RESULTS: Haplotype analysis showed a statistically significant association between some NOS3 gene variants and features of metabolic syndrome. Relative to the most common haplotype, 121, the haplotype 212 was associated with an increased odds ratio (OR) for metabolic syndrome [OR = 1.81, 95% confidence interval (CI) 1.15-2.84], and for decreased HDL-cholesterol concentrations (OR 1.52, 95% CI 1.01-2.29), and with increased mean values for the homeostasis model assessment of insulin resistance (P = 0.043), and triglycerides (P = 0.026). CONCLUSIONS: Our results suggest that genetic variation at the eNOS locus is associated with features of metabolic syndrome, and might represent a new genetic susceptibility component for insulin resistance, hypertriglyceridemia, and low HDL-cholesterol concentrations. PMID- 17110474 TI - Detection of epidermal growth factor receptor variations by partially denaturing HPLC. AB - BACKGROUND: Epidermal growth factor receptor gene (EGFR) variants may be useful markers for identifying responders to gefitinib and erlotinib, small-molecule tyrosine kinase inhibitors of EGFR; therefore, sensitive and cost-effective assays are needed to detect EGFR variants in routine clinical samples. We have developed a partially denaturing HPLC (pDHPLC) assay that is superior to direct sequencing with respect to detection limits, costs, and time requirements. METHODS: Primers, temperatures, and buffer conditions were optimized for PCR pDHPLC analysis of EGFR exons 18-21. We evaluated the detection limits of pDHPLC and direct sequencing by analyzing mixtures of wild-type and variant EGFR DNA and screened 192 lung cancer samples to examine the diversity of pDHPLC-detectable variants. To assess amenability to routine analysis, we tested lung and pleural tissue specimens from 14 lung cancer patients treated with gefitinib. RESULTS: The detection limits for variant alleles were 1:100 for pDHPLC and 1:5 for direct sequencing. pDHPLC analysis detected 26 unique EGFR variants, including the common deletions in exon 19 and substitutions in codons 787 and 858. Direct sequencing could not identify 30% (18 of 60) of the variant amplicons identified by pDHPLC. We identified these 18 amplicons by fraction collection after pDHPLC analysis. Analysis of a limited series of lung biopsy samples detected EGFR variants more frequently in gefitinib responders than in nonresponders. pDHPLC analysis was 56% less expensive and 39% faster than direct sequencing. CONCLUSIONS: pDHPLC-based analysis detects EGFR variations in routine clinical samples with a better detection limit and lower cost and time requirement than direct sequencing. PMID- 17110475 TI - High-volume extraction of nucleic acids by magnetic bead technology for ultrasensitive detection of bacteria in blood components. AB - BACKGROUND: Nucleic acid isolation, the most technically demanding and laborious procedure performed in molecular diagnostics, harbors the potential for improvements in automation. A recent development is the use of magnetic beads covered with nucleic acid-binding matrices. We adapted this technology with a broad-range 23S rRNA real-time reverse transcription (RT)-PCR assay for fast and sensitive detection of bacterial contamination of blood products. METHODS: We investigated different protocols for an automated high-volume extraction method based on magnetic-separation technology for the extraction of bacterial nucleic acids from platelet concentrates (PCs). We added 2 model bacteria, Staphylococcus epidermidis and Escherichia coli, to a single pool of apheresis-derived, single donor platelets and assayed the PCs by real-time RT-PCR analysis with an improved primer-probe system and locked nucleic acid technology. Co-amplification of human beta(2)-microglobulin mRNA served as an internal control (IC). We used probit analysis to calculate the minimum concentration of bacteria that would be detected with 95% confidence. RESULTS: For automated magnetic bead-based extraction technology with the real-time RT-PCR, the 95% detection limit was 29 x 10(3) colony-forming units (CFU)/L for S. epidermidis and 22 x 10(3) CFU/L for E. coli. No false-positive results occurred, either due to nucleic acid contamination of reagents or externally during testing of 1030 PCs. CONCLUSIONS: High-volume nucleic acid extraction improved the detection limit of the assay. The improvement of the primer-probe system and the integration of an IC make the RT-PCR assay appropriate for bacteria screening of platelets. PMID- 17110476 TI - A modified algorithm for the improvement of composite interval mapping. AB - Composite interval mapping (CIM) is the most commonly used method for mapping quantitative trait loci (QTL) with populations derived from biparental crosses. However, the algorithm implemented in the popular QTL Cartographer software may not completely ensure all its advantageous properties. In addition, different background marker selection methods may give very different mapping results, and the nature of the preferred method is not clear. A modified algorithm called inclusive composite interval mapping (ICIM) is proposed in this article. In ICIM, marker selection is conducted only once through stepwise regression by considering all marker information simultaneously, and the phenotypic values are then adjusted by all markers retained in the regression equation except the two markers flanking the current mapping interval. The adjusted phenotypic values are finally used in interval mapping (IM). The modified algorithm has a simpler form than that used in CIM, but a faster convergence speed. ICIM retains all advantages of CIM over IM and avoids the possible increase of sampling variance and the complicated background marker selection process in CIM. Extensive simulations using two genomes and various genetic models indicated that ICIM has increased detection power, a reduced false detection rate, and less biased estimates of QTL effects. PMID- 17110477 TI - SSP2 and OSW1, two sporulation-specific genes involved in spore morphogenesis in Saccharomyces cerevisiae. AB - Spore formation in Saccharomyces cerevisiae requires the synthesis of prospore membranes (PSMs) followed by the assembly of spore walls (SWs). We have characterized extensively the phenotypes of mutants in the sporulation-specific genes, SSP2 and OSW1, which are required for spore formation. A striking feature of the osw1 phenotype is asynchrony of spore development, with some spores displaying defects in PSM formation and others spores in the same ascus blocked at various stages in SW development. The Osw1 protein localizes to spindle pole bodies (SPBs) during meiotic nuclear division and subsequently to PSMs/SWs. We propose that Osw1 performs a regulatory function required to coordinate the different stages of spore morphogenesis. In the ssp2 mutant, nuclei are surrounded by PSMs and SWs; however, PSMs and SWs often also encapsulate anucleate bodies both inside and outside of spores. In addition, the SW is not as thick as in wild type. The ssp2 mutant defect is partially suppressed by overproduction of either Spo14 or Sso1, both of which promote the fusion of vesicles at the outer plaque of the SPB early in PSM formation. We propose that Ssp2 plays a role in vesicle fusion during PSM formation. PMID- 17110478 TI - Drosophila melanogaster male somatic cells feminized solely by TraF can collaborate with female germ cells to make functional eggs. AB - Female differentiation of Drosophila germ cells is induced by cell-nonautonomous signals generated in the gonadal soma that work with germ-cell-autonomous signals determined by germ-cell X chromosome dose. Generation of the nonautonomous feminizing signals was known to involve female-specific protein encoded by the master sex-determination gene Sex-lethal (Sxl) acting on its switch-gene target transformer (tra) to produce Tra(F) protein. However, it was not known whether Sxl's action on tra alone would suffice to trigger a fully feminizing nonautonomous signal. We developed a constitutively feminizing tra transgene that allowed us to answer this question. In gynanders (XX//XO mosaics) feminized by this Tra(F) transgene, functionally Sxl- haplo-X (chromosomally male) somatic cells collaborated successfully with diplo-X (chromosomally female) germ cells to make functional eggs. The fertility of such gynanders shows not only that Tra(F) is sufficient to elicit a fully feminizing nonautonomous signal, but also that haplo-X somatic cells can execute all other somatic functions required for oogenesis, despite the fact that their genome is not expected to be dosage compensated for such diplo-X-specific functions. The unexpected observation that some Tra(F)-feminized gynanders failed to lay their eggs showed there to be diplo X cells outside the gonad for which Tra(F)-feminized haplo-X cells cannot substitute. PMID- 17110479 TI - Teaching how to prepare a manuscript by means of rewriting published scientific papers. AB - The objective of the course described here is to train undergraduate students to write a scientific manuscript. The students participate in selection of a current topic in contemporary genetics or molecular biology by choosing the most interesting paper of a given year. After the teacher provides all essential background about the selected subject, he or she selects a recent article from the corresponding field and presents the students with all the necessary information contained in the paper without revealing its title and author. The data presented in the paper are reviewed by the class so that each student understands every experiment and the main points of the work. Simultaneously, the students are informed about the basic rules of writing the individual sections of a scientific paper. They are then asked to write and formally submit a manuscript summarizing the data. Finally, the students act as reviewers of their colleagues' manuscripts and compare their papers with the original published articles. This enables them to identify both the strengths and the weaknesses of their manuscripts and to gain confidence in the writing skills that will be so critical for their future scientific activities. PMID- 17110480 TI - Accumulation of deleterious mutations in small abiotic populations of RNA. AB - The accumulation of slightly deleterious mutations in populations leads to the buildup of a genetic load and can cause the extinction of populations of small size. Mutation-accumulation experiments have been used to study this process in a wide variety of organisms, yet the exact mutational underpinnings of genetic loads and their fitness consequences remain poorly characterized. Here, we use an abiotic system of RNA populations evolving continuously in vitro to examine the molecular events that can instigate a genetic load. By tracking the fitness decline of ligase ribozyme populations with bottleneck sizes between 100 and 3000 molecules, we detected the appearance and subsequent fixation of both slightly deleterious mutations and advantageous mutations. Smaller populations went extinct in significantly fewer generations than did larger ones, supporting the notion of a mutational meltdown. These data suggest that mutation accumulation was an important evolutionary force in the prebiotic RNA world and that mechanisms such as recombination to ameliorate genetic loads may have been in place early in the history of life. PMID- 17110481 TI - Ecological zones rather than molecular forms predict genetic differentiation in the malaria vector Anopheles gambiae s.s. in Ghana. AB - The malaria mosquito Anopheles gambiae s.s. is rapidly becoming a model for studies on the evolution of reproductive isolation. Debate has centered on the taxonomic status of two forms (denoted M and S) within the nominal taxon identified by point mutations in the X-linked rDNA region. Evidence is accumulating that there are significant barriers to gene flow between these forms, but that the barriers are not complete throughout the entire range of their distribution. We sampled populations from across Ghana and southern Burkina Faso, West Africa, from areas where the molecular forms occurred in both sympatry and allopatry. Neither Bayesian clustering methods nor F(ST)-based analysis of microsatellite data found differentiation between the M and S molecular forms, but revealed strong differentiation among different ecological zones, irrespective of M/S status and with no detectable effect of geographical distance. Although no M/S hybrids were found in the samples, admixture analysis detected evidence of contemporary interform gene flow, arguably most pronounced in southern Ghana where forms occur sympatrically. Thus, in the sampled area of West Africa, lack of differentiation between M and S forms likely reflects substantial introgression, and ecological barriers appear to be of greater importance in restricting gene flow. PMID- 17110482 TI - The Saccharomyces cerevisiae ATP22 gene codes for the mitochondrial ATPase subunit 6-specific translation factor. AB - Mutations in the Saccharomyces cerevisiae ATP22 gene were previously shown to block assembly of the F0 component of the mitochondrial proton-translocating ATPase. Further inquiries into the function of Atp22p have revealed that it is essential for translation of subunit 6 of the mitochondrial ATPase. The mutant phenotype can be partially rescued by the presence in the same cell of wild-type mitochondrial DNA and a rho- deletion genome in which the 5'-UTR, first exon, and first intron of COX1 are fused to the fourth codon of ATP6. The COX1/ATP6 gene is transcribed and processed to the mature mRNA by splicing of the COX1 intron from the precursor. The hybrid protein translated from the novel mRNA is proteolytically cleaved at the normal site between residues 10 and 11 of the subunit 6 precursor, causing the release of the polypeptide encoded by the COX1 exon. The ability of the rho- suppressor genome to express subunit 6 in an atp22 null mutant constitutes strong evidence that translation of subunit 6 depends on the interaction of Atp22p with the 5'-UTR of the ATP6 mRNA. PMID- 17110483 TI - A genetic screen in Drosophila for genes interacting with senseless during neuronal development identifies the importin moleskin. AB - Senseless (Sens) is a conserved transcription factor required for normal development of the Drosophila peripheral nervous system. In the Drosophila retina, sens is necessary and sufficient for differentiation of R8 photoreceptors and interommatidial bristles (IOBs). When Sens is expressed in undifferentiated cells posterior to the morphogenetic furrow, ectopic IOBs are formed. This phenotype was used to identify new members of the sens pathway in a dominant modifier screen. Seven suppressor and three enhancer complementation groups were isolated. Three groups from the screen are the known genes Delta, lilliputian, and moleskin/DIM-7 (msk), while the remaining seven groups represent novel genes with previously undefined functions in neural development. The nuclear import gene msk was identified as a potent suppressor of the ectopic interommatidial bristle phenotype. In addition, msk mutant adult eyes are extremely disrupted with defects in multiple cell types. Reminiscent of the sens mutant phenotype, msk eyes demonstrate reductions in the number of R8 photoreceptors due to an R8 to R2,5 fate switch, providing genetic evidence that Msk is a component of the sens pathway. Interestingly, in msk tissue, the loss of R8 fate occurs earlier than with sens and suggests a previously unidentified stage of R8 development between atonal and sens. PMID- 17110484 TI - Chi hotspot activity in Escherichia coli without RecBCD exonuclease activity: implications for the mechanism of recombination. AB - The major pathway of genetic recombination and DNA break repair in Escherichia coli requires RecBCD enzyme, a complex nuclease and DNA helicase regulated by Chi sites (5'-GCTGGTGG-3'). During its unwinding of DNA containing Chi, purified RecBCD enzyme has two alternative nucleolytic reactions, depending on the reaction conditions: simple nicking of the Chi-containing strand at Chi or switching of nucleolytic degradation from the Chi-containing strand to its complement at Chi. We describe a set of recC mutants with a novel intracellular phenotype: retention of Chi hotspot activity in genetic crosses but loss of detectable nucleolytic degradation as judged by the growth of mutant T4 and lambda phages and by assay of cell-free extracts. We conclude that RecBCD enzyme's nucleolytic degradation of DNA is not necessary for intracellular Chi hotspot activity and that nicking of DNA by RecBCD enzyme at Chi is sufficient. We discuss the bearing of these results on current models of RecBCD pathway recombination. PMID- 17110485 TI - Cytogenetic and molecular characterization of heterochromatin gene models in Drosophila melanogaster. AB - In the past decade, genome-sequencing projects have yielded a great amount of information on DNA sequences in several organisms. The release of the Drosophila melanogaster heterochromatin sequence by the Drosophila Heterochromatin Genome Project (DHGP) has greatly facilitated studies of mapping, molecular organization, and function of genes located in pericentromeric heterochromatin. Surprisingly, genome annotation has predicted at least 450 heterochromatic gene models, a figure 10-fold above that defined by genetic analysis. To gain further insight into the locations and functions of D. melanogaster heterochromatic genes and genome organization, we have FISH mapped 41 gene models relative to the stained bands of mitotic chromosomes and the proximal divisions of polytene chromosomes. These genes are contained in eight large scaffolds, which together account for approximately 1.4 Mb of heterochromatic DNA sequence. Moreover, developmental Northern analysis showed that the expression of 15 heterochromatic gene models tested is similar to that of the vital heterochromatic gene Nipped-A, in that it is not limited to specific stages, but is present throughout all development, despite its location in a supposedly "silent" region of the genome. This result is consistent with the idea that genes resident in heterochromatin can encode essential functions. PMID- 17110486 TI - An ectopic expression screen reveals the protective and toxic effects of Drosophila seminal fluid proteins. AB - In Drosophila melanogaster, seminal fluid regulates the reproductive and immune responses of mated females. Some seminal fluid proteins may provide protective functions to mated females, such as antimicrobial activity and/or stimulation of antimicrobial gene expression levels, while others appear to have negative effects, contributing to a "cost of mating." To identify seminal proteins that could participate in these phenomena, we used a systemic ectopic expression screen to test the effects on unmated females of proteins normally produced by the male accessory gland (Acps). Of the 21 ectopically expressed Acps that we tested for ability to assist in the clearance of a bacterial infection with Serratia marcescens, 3 Acps significantly reduced the bacterial counts of infected females, suggesting a protective role. Of the 23 Acps that we tested for toxicity, 3 were toxic, including one that has been implicated in the cost of mating in another study. We also tested ectopic expression females for other Acp induced effects, but found no additional Acps that affected egg laying or receptivity upon ectopic expression. PMID- 17110487 TI - Temporal estimates of effective population size in species with overlapping generations. AB - The standard temporal method for estimating effective population size (N(e)) assumes that generations are discrete, but it is routinely applied to species with overlapping generations. We evaluated bias in the estimates N(e) caused by violation of this assumption, using simulated data for three model species: humans (type I survival), sparrow (type II), and barnacle (type III). We verify a previous proposal by Felsenstein that weighting individuals by reproductive value is the correct way to calculate parametric population allele frequencies, in which case the rate of change in age-structured populations conforms to that predicted by discrete-generation models. When the standard temporal method is applied to age-structured species, typical sampling regimes (sampling only newborns or adults; randomly sampling the entire population) do not yield properly weighted allele frequencies and result in biased N(e). The direction and magnitude of the bias are shown to depend on the sampling method and the species' life history. Results for populations that grow (or decline) at a constant rate paralleled those for populations of constant size. If sufficient demographic data are available and certain sampling restrictions are met, the Jorde-Ryman modification of the temporal method can be applied to any species with overlapping generations. Alternatively, spacing the temporal samples many generations apart maximizes the drift signal compared to sampling biases associated with age structure. PMID- 17110488 TI - Mutations leading to loss of sporulation ability in Bacillus subtilis are sufficiently frequent to favor genetic canalization. AB - We measured the rate of mutations impairing sporulation ability in Bacillus subtilis as 0.003 in a mutator population, following 6000 generations of strong selection for sporulation that have previously been described. This means that the product of the population size and the functional mutation rate is approximately 10(5), well within the parameter range for which genetic canalization of sporulation ability is expected. PMID- 17110489 TI - Controlling the false-positive rate in multilocus genome scans for selection. AB - Rapid typing of genetic variation at many regions of the genome is an efficient way to survey variability in natural populations in an effort to identify segments of the genome that have experienced recent natural selection. Following such a genome scan, individual regions may be chosen for further sequencing and a more detailed analysis of patterns of variability, often to perform a parametric test for selection and to estimate the strength of a recent selective sweep. We show here that not accounting for the ascertainment of loci in such analyses leads to false inference of natural selection when the true model is selective neutrality, because the procedure of choosing unusual loci (in comparison to the rest of the genome-scan data) selects regions of the genome with genealogies similar to those expected under models of recent directional selection. We describe a simple and efficient correction for this ascertainment bias, which restores the false-positive rate to near-nominal levels. For the parameters considered here, we find that obtaining a test with the expected distribution of P-values depends on accurately accounting both for ascertainment of regions and for demography. Finally, we use simulations to explore the utility of relying on outlier loci to detect recent selective sweeps. We find that measures of diversity and of population differentiation are more effective than summaries of the site-frequency spectrum and that sequencing larger regions (2.5 kbp) in genome-scan studies leads to more power to detect recent selective sweeps. PMID- 17110490 TI - Nearly identical paralogs: implications for maize (Zea mays L.) genome evolution. AB - As an ancient segmental tetraploid, the maize (Zea mays L.) genome contains large numbers of paralogs that are expected to have diverged by a minimum of 10% over time. Nearly identical paralogs (NIPs) are defined as paralogous genes that exhibit > or = 98% identity. Sequence analyses of the "gene space" of the maize inbred line B73 genome, coupled with wet lab validation, have revealed that, conservatively, at least approximately 1% of maize genes have a NIP, a rate substantially higher than that in Arabidopsis. In most instances, both members of maize NIP pairs are expressed and are therefore at least potentially functional. Of evolutionary significance, members of many NIP families also exhibit differential expression. The finding that some families of maize NIPs are closely linked genetically while others are genetically unlinked is consistent with multiple modes of origin. NIPs provide a mechanism for the maize genome to circumvent the inherent limitation that diploid genomes can carry at most two "alleles" per "locus." As such, NIPs may have played important roles during the evolution and domestication of maize and may contribute to the success of long term selection experiments in this important crop species. PMID- 17110491 TI - Antagonizing scalloped with a novel vestigial construct reveals an important role for scalloped in Drosophila melanogaster leg, eye and optic lobe development. AB - Scalloped (SD), a TEA/ATTS-domain-containing protein, is required for the proper development of Drosophila melanogaster. Despite being expressed in a variety of tissues, most of the work on SD has been restricted to understanding its role and function in patterning the adult wing. To gain a better understanding of its role in development, we generated sd(47M) flip-in mitotic clones. The mitotic clones had developmental defects in the leg and eye. Further, by removing the VG domains involved in activation, we created a reagent (VGDeltaACT) that disrupts the ability of SD to form a functional transcription factor complex and produced similar phenotypes to the flip-in mitotic clones. The VGDeltaACT construct also disrupted adult CNS development. Expression of the VGDeltaACT construct in the wing alters the cellular localization of VG and produces a mutant phenotype, indicating that the construct is able to antagonize the normal function of the SD/VG complex. Expression of the protein:protein interaction portion of SD is also able to elicit similar phenotypes, suggesting that SD interacts with other cofactors in the leg, eye, and adult CNS. Furthermore, antagonizing SD in larval tissues results in cell death, indicating that SD may also have a role in cell survival. PMID- 17110492 TI - Fine mapping reveals sex bias in quantitative trait loci affecting growth, skeletal size and obesity-related traits on mouse chromosomes 2 and 11. AB - Previous speed congenic analysis has suggested that the expression of growth and obesity quantitative trait loci (QTL) on distal mouse chromosomes (MMU) 2 and 11, segregating between the CAST/EiJ (CAST) and C57BL/6J-hg/hg (HG) strains, is dependent on sex. To confirm, fine map, and further evaluate QTL x sex interactions, we constructed congenic by recipient F2 crosses for the HG.CAST (D2Mit329-D2Mit457)N(6) (HG2D) and HG.CAST-(D11Mit260-D11Mit255)N(6) (HG11) congenic strains. Over 700 F2 mice were densely genotyped and phenotyped for a panel of 40 body and organ weight, skeletal length, and obesity-related traits at 9 weeks of age. Linkage analysis revealed 20 QTL affecting a representative subset of phenotypes in HG2DF2 and HG11F2 mice. The effect of sex was quantified by comparing two linear models: the first model included sex as an additive covariate and the second incorporated sex as an additive and an interactive covariate. Of the 20 QTL, 8 were sex biased, sex specific, or sex antagonistic. Most traits were regulated by single QTL; however, two closely linked loci were identified for five traits in HG2DF2 mice. Additionally, the confidence intervals for most QTL were significantly reduced relative to the original mapping results, setting the stage for quantitative trait gene (QTG) discovery. These results highlight the importance of assessing the contribution of sex in complex trait analyses. PMID- 17110493 TI - Multilevel selection 2: Estimating the genetic parameters determining inheritance and response to selection. AB - Interactions among individuals are universal, both in animals and in plants and in natural as well as domestic populations. Understanding the consequences of these interactions for the evolution of populations by either natural or artificial selection requires knowledge of the heritable components underlying them. Here we present statistical methodology to estimate the genetic parameters determining response to multilevel selection of traits affected by interactions among individuals in general populations. We apply these methods to obtain estimates of genetic parameters for survival days in a population of layer chickens with high mortality due to pecking behavior. We find that heritable variation is threefold greater than that obtained from classical analyses, meaning that two-thirds of the full heritable variation is hidden to classical analysis due to social interactions. As a consequence, predicted responses to multilevel selection applied to this population are threefold greater than classical predictions. This work, combined with the quantitative genetic theory for response to multilevel selection presented in an accompanying article in this issue, enables the design of selection programs to effectively reduce competitive interactions in livestock and plants and the prediction of the effects of social interactions on evolution in natural populations undergoing multilevel selection. PMID- 17110494 TI - Multilevel selection 1: Quantitative genetics of inheritance and response to selection. AB - Interaction among individuals is universal, both in animals and in plants, and substantially affects evolution of natural populations and responses to artificial selection in agriculture. Although quantitative genetics has successfully been applied to many traits, it does not provide a general theory accounting for interaction among individuals and selection acting on multiple levels. Consequently, current quantitative genetic theory fails to explain why some traits do not respond to selection among individuals, but respond greatly to selection among groups. Understanding the full impacts of heritable interactions on the outcomes of selection requires a quantitative genetic framework including all levels of selection and relatedness. Here we present such a framework and provide expressions for the response to selection. Results show that interaction among individuals may create substantial heritable variation, which is hidden to classical analyses. Selection acting on higher levels of organization captures this hidden variation and therefore always yields positive response, whereas individual selection may yield response in the opposite direction. Our work provides testable predictions of response to multilevel selection and reduces to classical theory in the absence of interaction. Statistical methodology provided elsewhere enables empirical application of our work to both natural and domestic populations. PMID- 17110495 TI - Genes affecting cell competition in Drosophila. AB - Cell competition is a homeostatic mechanism that regulates the size attained by growing tissues. We performed an unbiased genetic screen for mutations that permit the survival of cells being competed due to haplo-insufficiency for RpL36. Mutations that protect RpL36 heterozygous clones include the tumor suppressors expanded, hippo, salvador, mats, and warts, which are members of the Warts pathway, the tumor suppressor fat, and a novel tumor-suppressor mutation. Other hyperplastic or neoplastic mutations did not rescue RpL36 heterozygous clones. Most mutations that rescue cell competition elevated Dpp-signaling activity, and the Dsmurf mutation that elevates Dpp signaling was also hyperplastic and rescued. Two nonlethal, nonhyperplastic mutations prevent the apoptosis of Minute heterozygous cells and suggest an apoptosis pathway for cell competition . In addition to rescuing RpL36 heterozygous cells, mutations in Warts pathway genes were supercompetitors that could eliminate wild-type cells nearby. The findings show that differences in Warts pathway activity can lead to competition and implicate the Warts pathway, certain other tumor suppressors, and novel cell death components in cell competition, in addition to the Dpp pathway implicated by previous studies. We suggest that cell competition might occur during tumor development in mammals. PMID- 17110496 TI - Thermodynamics of neutral protein evolution. AB - Naturally evolving proteins gradually accumulate mutations while continuing to fold to stable structures. This process of neutral evolution is an important mode of genetic change and forms the basis for the molecular clock. We present a mathematical theory that predicts the number of accumulated mutations, the index of dispersion, and the distribution of stabilities in an evolving protein population from knowledge of the stability effects (delta deltaG values) for single mutations. Our theory quantitatively describes how neutral evolution leads to marginally stable proteins and provides formulas for calculating how fluctuations in stability can overdisperse the molecular clock. It also shows that the structural influences on the rate of sequence evolution observed in earlier simulations can be calculated using just the single-mutation delta deltaG values. We consider both the case when the product of the population size and mutation rate is small and the case when this product is large, and show that in the latter case the proteins evolve excess mutational robustness that is manifested by extra stability and an increase in the rate of sequence evolution. All our theoretical predictions are confirmed by simulations with lattice proteins. Our work provides a mathematical foundation for understanding how protein biophysics shapes the process of evolution. PMID- 17110497 TI - Linkage maps of the dwarf and Normal lake whitefish (Coregonus clupeaformis) species complex and their hybrids reveal the genetic architecture of population divergence. AB - Elucidating the genetic architecture of population divergence may reveal the evolution of reproductive barriers and the genomic regions implicated in the process. We assembled genetic linkage maps for the dwarf and Normal lake whitefish species complex and their hybrids. A total of 877 AFLP loci and 30 microsatellites were positioned. The homology of mapped loci between families supported the existence of 34 linkage groups (of 40n expected) exhibiting 83% colinearity among linked loci between these two families. Classes of AFLP markers were not randomly distributed among linkage groups. Both AFLP and microsatellites exhibited deviations from Mendelian expectations, with 30.4% exhibiting significant segregation distortion across 28 linkage groups of the four linkage maps in both families (P < 0.00001). Eight loci distributed over seven homologous linkage groups were significantly distorted in both families and the level of distortion, when comparing homologous loci of the same phase between families, was correlated (Spearman R = 0.378, P = 0.0021). These results suggest that substantial divergence incurred during allopatric glacial separation and subsequent sympatric ecological specialization has resulted in several genomic regions that are no longer complementary between dwarf and Normal populations issued from different evolutionary glacial lineages. PMID- 17110498 TI - Association genetics in Pinus taeda L. I. Wood property traits. AB - Genetic association is a powerful method for dissecting complex adaptive traits due to (i) fine-scale mapping resulting from historical recombination, (ii) wide coverage of phenotypic and genotypic variation within a single experiment, and (iii) the simultaneous discovery of loci and alleles. In this article, genetic association among single nucleotide polymorphisms (58 SNPs) from 20 wood- and drought-related candidate genes and an array of wood property traits with evolutionary and commercial importance, namely, earlywood and latewood specific gravity, percentage of latewood, earlywood microfibril angle, and wood chemistry (lignin and cellulose content), was tested using mixed linear models (MLMs) that account for relatedness among individuals by using a pairwise kinship matrix. Population structure, a common systematic bias in association studies, was assessed using 22 nuclear microsatellites. Different phenotype:genotype associations were found, some of them confirming previous evidence from collocation of QTL and genes in linkage maps (for example, 4cl and percentage of latewood) and two that involve nonsynonymous polymorphisms (cad SNP M28 with earlywood specific gravity and 4cl SNP M7 with percentage of latewood). The strongest genetic association found in this study was between allelic variation in alpha-tubulin, a gene involved in the formation of cortical microtubules, and earlywood microfibril angle. Intragenic LD decays rapidly in conifers; thus SNPs showing genetic association are likely to be located in close proximity to the causative polymorphisms. This first multigene association genetic study in forest trees has shown the feasibility of candidate gene strategies for dissecting complex adaptive traits, provided that genes belonging to key pathways and appropriate statistical tools are used. This approach is of particular utility in species such as conifers, where genomewide strategies are limited by their large genomes. PMID- 17110499 TI - Rates of recombination in the ribosomal DNA of apomictically propagated Daphnia obtusa lines. AB - Ribosomal (r)DNA undergoes concerted evolution, the mechanisms of which are unequal crossing over and gene conversion. Despite the fundamental importance of these mechanisms to the evolution of rDNA, their rates have been estimated only in a few model species. We estimated recombination rate in rDNA by quantifying the relative frequency of intraindividual length variants in an expansion segment of the 18S rRNA gene of the cladoceran crustacean, Daphnia obtusa, in four apomictically propagated lines. We also used quantitative PCR to estimate rDNA copy number. The apomictic lines were sampled every 5 generations for 90 generations, and we considered each significant change in the frequency distribution of length variants between time intervals to be the result of a recombination event. Using this method, we calculated the recombination rate for this region to be 0.02-0.06 events/generation on the basis of three different estimates of rDNA copy number. In addition, we observed substantial changes in rDNA copy number within and between lines. Estimates of haploid copy number varied from 53 to 233, with a mean of 150. We also measured the relative frequency of length variants in 30 lines at generations 5, 50, and 90. Although length variant frequencies changed significantly within and between lines, the overall average frequency of each length variant did not change significantly between the three generations sampled, suggesting that there is little or no bias in the direction of change due to recombination. PMID- 17110500 TI - Kinins promote B2 receptor endocytosis and delay constitutive B1 receptor endocytosis. AB - Upon sustained insult, kinins are released and many kinin responses, such as inflammatory pain, adapt from a B2 receptor (B2R) type in the acute phase to a B1 receptor (B1R) type in the chronic phase. In this study, we show that kinins modulate receptor endocytosis to rapidly decrease B2R and increase B1R on the cell surface. B2Rs, which require agonist for activity, are stable plasma membrane components without agonist but recruit beta-arrestin 2, internalize in a clathrin-dependent manner, and recycle rapidly upon agonist treatment. In contrast, B1Rs, which are inducible and constitutively active, constitutively internalize without agonist via a clathrin-dependent pathway, do not recruit beta arrestin 2, bind G protein-coupled receptor sorting protein, and target lysosomes for degradation. Agonist delays B1R endocytosis, thus transiently stabilizing the receptor. Most of the receptor trafficking phenotypes are transplantable from one receptor to the other through exchange of the C-terminal receptor tails, indicating that the tails contain epitopes that are important for the binding of protein partners that participate in the endocytic and postendocytic receptor choices. It is noteworthy that the agonist delay of B1R endocytosis is not transplanted to the B2R via the B1R tail, suggesting that this property of the B1R requires another domain. These events provide a rapid kinin-dependent mechanism for 1) regulating the constitutive B1R activity and 2) shifting the balance of accessible receptors in favor of B1R. PMID- 17110501 TI - Functional involvement of multidrug resistance-associated protein 4 (MRP4/ABCC4) in the renal elimination of the antiviral drugs adefovir and tenofovir. AB - Acyclic nucleotide phosphonates (adefovir, cidofovir, and tenofovir) are eliminated predominantly into the urine, and renal failure is their dose-limiting toxicity, particularly for adefovir and cidofovir. In this study, we examined the involvement of multidrug resistance-associated protein (MRP)4 (ABCC4) in their luminal efflux in the kidney. ATP-dependent uptake of adefovir and tenofovir but not cidofovir was observed only in the membrane vesicles expressing MRP4. The ATP dependent uptake of adefovir and tenofovir by MRP4 was not saturated at 1 mM. The ATP-dependent uptake of adefovir by membrane vesicles expressing MRP4 was osmotic sensitive. No ATP-dependent uptake of either agent was observed in the membrane vesicles expressing human MRP2 or breast cancer resistance protein. These nucleotide analogs were given to mice by constant intravenous infusion, and the plasma, urine, and tissue concentrations were determined. The kidney accumulation of adefovir and tenofovir was significantly greater in Mrp4 knockout mice (130 versus 66 and 191 versus 87 pmol/g tissue, respectively); thus, the renal luminal efflux clearance was estimated to be 37 and 46%, respectively, of the control. There was no difference in the fraction of mono- and diphosphorylated forms of adefovir in the kidney between wild-type and Mrp4 knockout mice. In mice, cidofovir was also eliminated via the urine by tubular secretion as well as glomerular filtration. There was no change in the kinetic parameters of cidofovir in Mrp4 knockout mice. Our results suggest that MRP4 is involved in the luminal efflux of both adefovir and tenofovir, but it makes only a limited contribution to the urinary excretion of cidofovir. PMID- 17110502 TI - Imino sugars are potent agonists of the human glucose sensor SGLT3. AB - Imino sugars are used to treat type 2 diabetes mellitus [miglitol (Glyset)] and lysosomal storage disorders [miglustat (Zavesca)] based on the inhibition of alpha-glucosidases and glucosyltransferases. In this substrate specificity study, we examined the interactions of imino sugars with a novel human glucose sensor, sodium/glucose cotransporter type 3 (hSGLT3), using expression in Xenopus laevis oocytes and electrophysiology. The results for hSGLT3 are compared with those for alpha-glucosidases and human SGLT type 1 (hSGLT1), a well characterized sodium/glucose cotransporter of the SGLT family. In general, substrates have lower apparent affinities (K0.5) for hSGLT3 than hSGLT1 (D-glucose, alpha-methyl D-glucose, 1-deoxy-D-glucose, and 4-deoxy-4-fluoro-D-glucose exhibit K0.5 values of 19, 21, 43, and 17 mM, respectively, for hSGLT3, and 0.5, 0.7, 10, and 0.07 mM, respectively, for hSGLT1). However, specificity of hSGLT3 binding is greater (D-galactose and 4-deoxy-4-fluoro-D-galactose are not hSGLT3 substrates, but have hSGLT1 K0.5 values of 0.6 and 1.3 mM). An important deviation from this trend is potent hSGLT3 activation by the imino sugars 1-deoxynojirimycin (DNJ), N hydroxylethyl-1-deoxynojirimycin (miglitol), N-butyl-1-deoxynojirimycin (miglustat), N-ethyl-1-deoxynojirimycin, and 1-deoxynojirimycin-1-sulfonic acid, with K0.5 values of 0.5 to 9 microM. The diastereomer 1-deoxygalactonojirimycin activates hSGT3 with a K0.5 value of 11 mM, a 3000-fold less potent interaction than is observed for DNJ (4 microM). These imino sugar binding characteristics are similar to those for alpha-glucosidases, but there are no interactions with hSGLT1. This work provides insights into hSGLT3 and -1 substrate binding interactions, establishes a pharmacological profile to study endogenous hSGLT3, and may have important ramifications for the clinical application of imino sugars. PMID- 17110503 TI - The classical Guyton view that mean systemic pressure, right atrial pressure, and venous resistance govern venous return is/is not correct. PMID- 17110504 TI - Characteristics of airflow in a CT-based ovine lung: a numerical study. AB - The transient airflow in a rigid, asymmetric monopodial sheep (ovine) tracheobronchial tree of up to 13 generations was investigated numerically. The lung geometry was segmented and reconstructed from computed-tomographic (CT) images. The flow characteristics in the image-based sheep airway were compared with the flow patterns produced by a Weibel-based model at prime locations. Boundary conditions were prescribed 1) a velocity profile from experimental data at the inlet and 2) zero pressure at the bronchial outlets. A mesh convergence study was carried out to establish confidence in the model predictions, and gross left-right ventilation was validated against experimental xenon wash-in-washout data. Detailed flow characteristics were investigated at three points in the breathing cycle: 1) peak inhalation, 2) peak exhalation, and 3) transition. Simulation results revealed fundamental differences between airflow in monopodial and bipodial branching airways. Compared with idealized bipodial flow, the flow in the sheep airway was asymmetric and highly vortical, especially during exhalation and transition. The streak lines during the inhalation phase suggest that the left and right upper lobes are ventilated by airflow in the peripheral region of the trachea. This work may contribute to understanding the interplay between structure and function in the lung. PMID- 17110505 TI - Pentoxifylline attenuation of experimental hepatopulmonary syndrome. AB - Hepatopulmonary syndrome (HPS) following rat common bile duct ligation results from pulmonary molecular changes that may be influenced by circulating TNF-alpha and increased vascular shear stress, through activation of NF-kappaB or Akt. Increased pulmonary microvascular endothelin B (ET(B)) receptor and endothelial nitric oxide synthase (eNOS) levels contribute to nitric oxide production and the development of experimental HPS. Pentoxifylline (PTX), a phosphodiesterase and nonspecific TNF-alpha inhibitor, ameliorates experimental HPS when begun before hepatic injury. However, how PTX influences the molecular events associated with initiation of experimental HPS after liver injury is established is unknown. We assessed the effects of PTX on the molecular and physiological features of HPS in vivo and on shear stress or TNF-alpha-mediated events in rat pulmonary microvascular endothelial cells in vitro. PTX significantly improved HPS without altering portal or systemic hemodynamics and downregulated pulmonary ET(B) receptor levels and eNOS expression and activation. These changes were associated with a reduction in circulating TNF levels and NF-kappaB activation and complete inhibition of Akt activation. In rat pulmonary microvascular endothelial cells, PTX inhibited shear stress-induced ET(B) receptor and eNOS expression and eNOS activation. These effects were also associated with inhibition of Akt activation and were reproduced by wortmanin. In contrast, TNF-alpha had no effects on endothelial ET(B) and eNOS alterations in vitro. PTX has direct effects in the pulmonary microvasculature, likely mediated through Akt inhibition, that ameliorate experimental HPS. PMID- 17110506 TI - Airway nitric oxide release is reduced after PBS inhalation in asthma. AB - Exhaled nitric oxide (NO) is elevated in asthma, but the underlying mechanisms remain poorly understood. Recent results in subjects with asthma have reported a decrease in exhaled breath pH and ammonia, as well as altered expression and activity of glutaminase in both alveolar and airway epithelial cells. This suggests that pH-dependent nitrite conversion to NO may be a source of exhaled NO in the asthmatic airway epithelium. However, the anatomic location (i.e., airway or alveolar region) of this pH-dependent NO release has not been investigated and could impact potential therapeutic strategies. We quantified airway (proximal) and alveolar (peripheral) contributions to exhaled NO at baseline and then after PBS inhalation in stable (mild-intermittent to severe) asthmatic subjects (20-44 yr old; n = 9) and healthy controls (22-41 yr old; n = 6). The mean (SD) maximum airway wall flux (pl/s) and alveolar concentration (ppb) at baseline in asthma subjects and healthy controls was 2,530 (2,572) and 5.42 (7.31) and 1,703 (1,567) and 1.88 (1.29), respectively. Compared with baseline, there is a significant decrease in the airway wall flux of NO in asthma as early as 15 min and continuing for up to 60 min (maximum -28% at 45 min) after PBS inhalation without alteration of alveolar concentration. Healthy control subjects did not display any changes in exhaled NO. We conclude that elevated airway NO at baseline in asthma is reduced by inhaled PBS. Thus airway NO may be, in part, due to nitrite conversion to NO and is consistent with airway pH dysregulation in asthma. PMID- 17110507 TI - Excessive heart rate response to orthostatic stress in postural tachycardia syndrome is not caused by anxiety. AB - Postural tachycardia syndrome (POTS) is characterized by excessive increases in heart rate (HR) without hypotension during orthostasis. The relationship between the tachycardia and anxiety is uncertain. Therefore, we tested whether the HR response to orthostatic stress in POTS is primarily related to psychological factors. POTS patients (n = 14) and healthy controls (n = 10) underwent graded venous pooling with lower body negative pressure (LBNP) to -40 mmHg while wearing deflated antishock trousers. "Sham" venous pooling was performed by 1) trouser inflation to 5 mmHg during LBNP and 2) vacuum pump activation without LBNP. HR responses to mental stress were also measured in both groups, and a questionnaire was used to measure psychological parameters. During LBNP, HR in POTS patients increased 39 +/- 5 beats/min vs. 19 +/- 3 beats/min in control subjects at -40 mmHg (P < 0.01). LBNP with trouser inflation markedly blunted the HR responses in the patients (9 +/- 2 beats/min) and controls (2 +/- 1 beats/min), and there was no HR increase during vacuum application without LBNP in either group. HR responses during mental stress were not different in the patients and controls (18 +/- 2 vs. 19 +/- 1 beats/min; P > 0.6). Anxiety, somatic vigilance, and catastrophic cognitions were significantly higher in the patients (P < 0.05), but they were not related to the HR responses during LBNP or mental stress (P > 0.1). These results suggest that the HR response to orthostatic stress in POTS patients is not caused by anxiety but that it is a physiological response that maintains arterial pressure during venous pooling. PMID- 17110508 TI - Androstenediol inhibits the trauma-hemorrhage-induced increase in caspase-3 by downregulating the inducible nitric oxide synthase pathway. AB - Soft tissue trauma and hemorrhage (T-H) diminishes various aspects of liver function, while it increases hepatic nitrate/nitrite, inducible nitric oxide synthase (iNOS), and endothelin-1 levels. Treatment with androstenediol (AED) inhibits the T-H-induced alterations of the above parameters. We sought to identify the molecular events underlying the beneficial effect of AED. Exposure of rats to T-H significantly increased the caspase-3 activity and protein, whereas treatment with AED significantly limited these increases. AED treatment also suppressed the T-H-induced increase in iNOS by effectively altering the levels of key transcription factors involved in the regulation of iNOS expression. Immunoprecipitation and immunoblotting analyses indicate that T-H increased apoptosome formation, and AED treatment significantly decreased it. Modulating the iNOS protein by transfecting cells with iNOS gene or small interfering RNA further confirmed the correlation between iNOS and caspase-3. Our data indicate that AED limits caspase-3 expression by suppressing the expression of transcription factors involved in the production of iNOS, resulting in decreased apoptosome. AED can potentially be a useful adjuvant for limiting liver apoptosis following T-H shock. PMID- 17110509 TI - Estimation of the functional role of arterial pathways to the buttock circulation during treadmill walking in patients with claudication. AB - The aim of the study was to estimate the functional contribution of the arterial inflow pathways to the pelvic circulation during walking in patients with stage 2 lower extremity arterial disease. Transcutaneous oxygen pressure (Ptc(O(2))) changes during exercise can be used to estimate the severity of regional blood flow impairment while walking. Seventy patients with stable lower limb claudication were studied using a multivariate linear regression model. The relationship between exercise-induced buttock Ptc(O(2)) changes, the ipsilateral calf Ptc(O(2)) changes, and the arterial diameters of the pelvic arteriographic pathways were analyzed. The ipsilateral hypogastric and lumbar pathway, as well as the ipsilateral calf Ptc(O(2)) changes, were the only variables significantly related to buttock Ptc(O(2)) changes (r = 0.47; P < 0.001). Their normalized respective contribution to the regressive model was 39%, 19%, and 18%. None of the contralateral hypogastric, mesenteric, and sacral pathways or pathways stemming from the external iliac artery showed significant correlation to buttock Ptc(O(2)) changes. The ipsilateral hypogastric and ipsilateral lumbar pathways are the major pathways responsible for the functional buttock blood flow supply during walking. The role of contralateral hypogastric, inferior mesenteric, and median sacral pathways and arteries distal to the internal iliac trunk is negligible in the normal or compensatory blood flow supply. Distal Ptc(O(2)) decrease at exercise aggravates proximal Ptc(O(2)) decrease, possibly through the occurrence of a "steal phenomenon" of distal over proximal circulation during walking. PMID- 17110510 TI - Transcranial Doppler estimation of cerebral blood flow and cerebrovascular conductance during modified rebreathing. AB - Clinical transcranial Doppler assessment of cerebral vasomotor reactivity (CVMR) uses linear regression of cerebral blood flow velocity (CBFV) vs. end-tidal CO(2) (Pet(CO(2))) under steady-state conditions. However, the cerebral blood flow (CBF)-Pet(CO(2)) relationship is nonlinear, even for moderate changes in CO(2). Moreover, CBF is increased by increases in arterial blood pressure (ABP) during hypercapnia. We used a modified rebreathing protocol to estimate CVMR during transient breath-by-breath changes in CBFV and Pet(CO(2)). Ten healthy subjects (6 men) performed 15 s of hyperventilation followed by 5 min of rebreathing, with supplemental O(2) to maintain arterial oxygen saturation constant. To minimize effects of changes in ABP on CVMR estimation, cerebrovascular conductance index (CVCi) was calculated. CBFV-Pet(CO(2)) and CVCi-Pet(CO(2)) relationships were quantified by both linear and nonlinear logistic regression. In three subjects, muscle sympathetic nerve activity was recorded. From hyperventilation to rebreathing, robust changes occurred in Pet(CO(2)) (20-61 Torr), CBFV (-44 to +104% of baseline), CVCi (-39 to +64%), and ABP (-19 to +23%) (all P < 0.01). Muscle sympathetic nerve activity increased by 446% during hypercapnia. The linear regression slope of CVCi vs. Pet(CO(2)) was less steep than that of CBFV (3 vs. 5%/Torr; P = 0.01). Logistic regression of CBF-Pet(CO(2)) (r(2) = 0.97) and CVCi-Pet(CO(2)) (r(2) = 0.93) was superior to linear regression (r(2) = 0.91, r(2) = 0.85; P = 0.01). CVMR was maximal (6-8%/Torr) for Pet(CO(2)) of 40-50 Torr. In conclusion, CBFV and CVCi responses to transient changes in Pet(CO(2)) can be described by a nonlinear logistic function, indicating that CVMR estimation varies within the range from hypocapnia to hypercapnia. Furthermore, quantification of the CVCi-Pet(CO(2)) relationship may minimize the effects of changes in ABP on the estimation of CVMR. The method developed provides insight into CVMR under transient breath-by-breath changes in CO(2). PMID- 17110511 TI - Oxidative stress contributes to chronic leg vasoconstriction in estrogen deficient postmenopausal women. AB - Basal whole leg blood flow and vascular conductance are reduced in estrogen deficient postmenopausal compared with premenopausal women. The underlying mechanisms are unknown, but oxidative stress could be involved. We studied 9 premenopausal [23 +/- 1 yr (mean +/- SE)] and 20 estrogen-deficient postmenopausal (55 +/- 1 yr) healthy women. During baseline control, oxidized low density lipoprotein (LDL), a marker of oxidative stress, was 50% greater in the postmenopausal women (P < 0.001). Basal whole leg blood flow (duplex ultrasound of femoral artery) was 34% lower in the postmenopausal women because of a 38% lower leg vascular conductance (P < 0.0001); mean arterial pressure was not different. Intravenous administration of a supraphysiological dose of the antioxidant ascorbic acid increased leg blood flow by 15% in the postmenopausal women as a result of an increase in leg vascular conductance (both P < 0.001), but it did not affect leg blood flow in premenopausal controls or mean arterial pressure in either group. In the pooled subjects, the changes in leg blood flow and leg vascular conductance with ascorbic acid were related to baseline plasma oxidized LDL (r = 0.46 and 0.53, P < 0.01) and waist-to-hip ratio and total body fat (r = 0.41-0.44, all P < 0.05). Our results are consistent with the hypothesis that oxidative stress contributes to chronic leg vasoconstriction and reduced basal whole leg blood flow in estrogen-deficient postmenopausal women. This oxidative stress-related suppression of leg vascular conductance and blood flow may be linked in part to increased total and abdominal adiposity. PMID- 17110512 TI - Decreased baroreflex sensitivity in acute schizophrenia. AB - Decreased vagal activity has been described in acute schizophrenia and might be associated with altered cardiovascular regulation and increased cardiac mortality. The aim of this study was to assess baroreflex sensitivity in the context of psychopathology. Twenty-one acute, psychotic, unmedicated patients with a diagnosis of paranoid schizophrenia were investigated after admission to the hospital. Results were compared with 21 healthy volunteers matched with respect to age and sex. Cardiovascular parameters obtained included measures for heart rate variability, baroreflex sensitivity, as well as cardiac output, left ventricular work index, and total peripheral resistance. All parameters investigated were analyzed using linear and novel nonlinear techniques. Positive and negative symptoms were assessed to estimate the impact of psychopathology on autonomic parameters. Subjects with acute schizophrenia showed reduction of baroreflex sensitivity accompanied by tachycardia and greatly increased left ventricular work index. Nonlinear parameters of baroreflex sensitivity correlated with positive symptoms. For heart rate variability, mainly parameters indicating parasympathetic modulation were decreased. Vascular pathology could be excluded as a confounding factor. These results reflect a dysfunctional cardiovascular regulation in acute schizophrenic patients at rest. The changes are similar to adaptational regulatory processes following stressful mental or physical tasks in healthy subjects. This study suggests that hyperarousal in acute schizophrenia is accompanied by decreased efferent vagal activity, thus increasing the risk for cardiovascular mortality. Future studies are warranted to examine the role of the sympathetic system and possible autonomic differences in hyperarousal induced by anxiety and/or external stressful events. PMID- 17110513 TI - Impact of endurance training on murine spontaneous activity, muscle mitochondrial DNA abundance, gene transcripts, and function. AB - We hypothesized that enhanced skeletal muscle mitochondrial function following aerobic exercise training is related to an increase in mitochondrial transcription factors, DNA abundance [mitochondrial DNA (mtDNA)], and mitochondria-related gene transcript levels, as well as spontaneous physical activity (SPA) levels. We report the effects of daily treadmill training on 12-wk old FVB mice for 5 days/wk over 8 wk at 80% peak O(2) consumption and studied the training effect on changes in body composition, glucose tolerance, muscle mtDNA muscle, mitochondria-related gene transcripts, in vitro muscle mitochondrial ATP production capacity (MATPC), and SPA levels. Compared with the untrained mice, the trained mice had higher peak O(2) consumption (+18%; P < 0.001), lower percentage of abdominal (-25.4%; P < 0.02) and body fat (-19.5%; P < 0.01), improved glucose tolerance (P < 0.04), and higher muscle mitochondrial enzyme activity (+19.5-43.8%; P < 0.04) and MATPC (+28.9 to +32.4%; P < 0.01). Gene array analysis showed significant differences in mRNAs of mitochondria-related ontology groups between the trained and untrained mice. Training also increased muscle mtDNA (+88.4 to +110%; P < 0.05), peroxisome proliferative-activated receptor-gamma coactivator-1alpha protein (+99.5%; P < 0.04), and mitochondrial transcription factor A mRNA levels (+21.7%; P < 0.004) levels. SPA levels were higher in trained mice (P = 0.056, two-sided t-test) and significantly correlated with two separate substrate-based measurements of MATPC (P < 0.02). In conclusion, aerobic exercise training enhances muscle mitochondrial transcription factors, mtDNA abundance, mitochondria-related gene transcript levels, and mitochondrial function, and this enhancement in mitochondrial function occurs in association with increased SPA. PMID- 17110514 TI - Transpulmonary pressures and lung mechanics with glossopharyngeal insufflation and exsufflation beyond normal lung volumes in competitive breath-hold divers. AB - Throughout life, most mammals breathe between maximal and minimal lung volumes determined by respiratory mechanics and muscle strength. In contrast, competitive breath-hold divers exceed these limits when they employ glossopharyngeal insufflation (GI) before a dive to increase lung gas volume (providing additional oxygen and intrapulmonary gas to prevent dangerous chest compression at depths recently greater than 100 m) and glossopharyngeal exsufflation (GE) during descent to draw air from compressed lungs into the pharynx for middle ear pressure equalization. To explore the mechanical effects of these maneuvers on the respiratory system, we measured lung volumes by helium dilution with spirometry and computed tomography and estimated transpulmonary pressures using an esophageal balloon after GI and GE in four competitive breath-hold divers. Maximal lung volume was increased after GI by 0.13-2.84 liters, resulting in volumes 1.5-7.9 SD above predicted values. The amount of gas in the lungs after GI increased by 0.59-4.16 liters, largely due to elevated intrapulmonary pressures of 52-109 cmH(2)O. The transpulmonary pressures increased after GI to values ranging from 43 to 80 cmH(2)O, 1.6-2.9 times the expected values at total lung capacity. After GE, lung volumes were reduced by 0.09-0.44 liters, and the corresponding transpulmonary pressures decreased to -15 to -31 cmH(2)O, suggesting closure of intrapulmonary airways. We conclude that the lungs of some healthy individuals are able to withstand repeated inflation to transpulmonary pressures far greater than those to which they would normally be exposed. PMID- 17110515 TI - Technical and physiological background of plasma volume measurement with indocyanine green: a clarification of misunderstandings. AB - The indocyanine green (ICG) dilution technique (DT) is frequently used for plasma volume (PV) measurement. However, because of inadequate knowledge about the properties of this dye, lack of accuracy has been attributed to the method. The aim of this report is to provide physiological background information about the ICG-DT to avoid some profound misunderstandings. When performing tracer dilution, one has to consider the tracer's distribution space before interpreting the result. For ICG, the distribution space is the total PV, i.e., circulating + noncirculating PV, fixed within the endothelial glycocalyx. The distribution space of red blood cells and large molecules, in contrast, is only the circulating part of PV. Therefore, it is erroneous to compare directly PV derived from different tracer dilution methods. The transcapillary escape rate of ICG should not relevantly influence measured PV if the method is performed properly, i.e., if a short time window of measurement is subjected to monoexponential extrapolation. A major problem of PV measurement in general is that the target itself is very inconstant. Thus, checking for constancy of ICG-DT with two consecutive measurements is unreliable. Nevertheless, the ICG-DT is a useful tool for determining PV, provided it is well understood by the investigator to enable correct interpretation of the results. PMID- 17110516 TI - Intermittent hyperthermia enhances skeletal muscle regrowth and attenuates oxidative damage following reloading. AB - Skeletal muscle reloading following disuse is characterized by profound oxidative damage. This study tested the hypothesis that intermittent hyperthermia during reloading attenuates oxidative damage and augments skeletal muscle regrowth following immobilization. Forty animals were randomly divided into four groups: control (Con), immobilized (Im), reloaded (RC), and reloaded and heated (RH). All groups but Con were immobilized for 7 days. Animals in the RC and RH groups were then reloaded for 7 days with (RH) or without (RC) hyperthermia (41-41.5 degrees C for 30 min on alternating days) during reloading. Heating resulted in approximately 25% elevation in heat shock protein expression (P < 0.05) and an approximately 30% greater soleus regrowth (P < 0.05) in RH compared with RC. Furthermore, oxidant damage was lower in the RH group compared with RC because nitrotyrosine and 4-hydroxy-2-nonenol were returned to near baseline when heating was combined with reloading. Reduced oxidant damage was independent of antioxidant enzymes (manganese superoxide dismutase, copper-zinc superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase). In summary, these data suggest that intermittent hyperthermia during reloading attenuates oxidative stress and improves the rate of skeletal muscle regrowth during reloading after immobilization. PMID- 17110517 TI - Bowman-Birk inhibitor concentrate prevents atrophy, weakness, and oxidative stress in soleus muscle of hindlimb-unloaded mice. AB - Antigravity muscles atrophy and weaken during prolonged mechanical unloading caused by bed rest or spaceflight. Unloading also induces oxidative stress in muscle, a putative cause of weakness. We tested the hypothesis that dietary supplementation with Bowman-Birk inhibitor concentrate (BBIC), a soy protein extract, would oppose these changes. Adult mice were fed a diet supplemented with 1% BBIC during hindlimb unloading for up to 12 days. Soleus muscles of mice fed the BBIC-supplemented diet weighed less, developed less force per cross-sectional area, and developed less total force after unloading than controls. BBIC supplementation was protective, blunting decrements in soleus muscle weight and force. Cytosolic oxidant activity was assessed using 2',7'-dichlorofluorescin diacetate. Oxidant activity increased in unloaded muscle, peaking at 3 days and remaining elevated through 12 days of unloading. Increases in oxidant activity correlated directly with loss of muscle mass and were abolished by BBIC supplementation. In vitro assays established that BBIC directly buffers reactive oxygen species and also inhibits serine protease activity. We conclude that dietary supplementation with BBIC protects skeletal muscle during prolonged unloading, promoting redox homeostasis in muscle fibers and blunting atrophy induced weakness. PMID- 17110518 TI - Evaluation of emphysema severity and progression in a rabbit model: comparison of hyperpolarized 3He and 129Xe diffusion MRI with lung morphometry. AB - The apparent diffusion coefficients (ADCs) of hyperpolarized (3)He and (129)Xe gases were measured in the lungs of rabbits with elastase-induced emphysema and correlated against the mean chord length from lung histology. In vivo measurements were performed at baseline and 2, 4, 6, and 8 wk after instillation of elastase (mild and moderate emphysema groups) or saline (control group). ADCs were determined from acquisitions that used two b values. To investigate the effect of b value on the results, b-value pairs of 0 and 1.6 s/cm(2) and 0 and 4.0 s/cm(2) were used for (3)He, and b-value pairs of 0 and 5.0 s/cm(2) and 0 and 10.0 s/cm(2) were used for (129)Xe. At 8 wk after instillation, the rabbits were euthanized, and the lungs were analyzed histologically and morphometrically. ADCs for the rabbits in the control group did not change significantly from baseline to week 8, whereas ADCs for the rabbits in the emphysema groups increased significantly (P < 0.05) for all gas and b-value combinations except (129)Xe with the b-value pair of 0 and 5.0 s/cm(2). The largest percent change in mean ADC from baseline to week 8 (15.3%) occurred with (3)He and the b-value pair of 0 and 1.6 s/cm(2) for rabbits in the moderate emphysema group. ADCs (all b values) were strongly correlated (r = 0.62-0.80, P < 0.001) with mean chord lengths from histology. These results further support the ability of diffusion-weighted MRI with hyperpolarized gases to detect regional and global structural changes of emphysema within the lung. PMID- 17110519 TI - Peripheral neuropathy does not alter the fractal dynamics of stride intervals of gait. AB - The purpose of this study was to determine the effect (if any) of significant sensory loss on the long-range correlations normally observed in the stride intervals of human gait. Fourteen patients with severe peripheral neuropathy and 12 gender-, age-, height-, and weight-matched nondiabetic controls participated. Subjects walked around an approximately 200-m open-level walkway for 10 min at their comfortable pace. Continuous knee joint kinematics were recorded and used to calculate a stride interval time series for each subject. Power spectral density and detrended fluctuation analyses were used to determine whether these stride intervals exhibited long-range correlations. If the loss of long-range correlations indicates deterioration of the central control of gait, then changes in peripheral sensation should have no effect. If instead the loss of long-range correlations is a consequence of a general inability to regulate gait cycle timing, then a similar loss should occur in patients with peripheral locomotor disorders. Both power spectral density analyses and detrended fluctuation analyses showed that temporal correlations in the stride times of neuropathic and control subjects were statistically identical (P = 0.954 and P = 0.974, respectively), despite slower gait speeds (P = 0.008) and increased stride time variability (P = 0.036) among the neuropathy patients. All subjects in both groups exhibited long-range correlations. These findings demonstrate that the normal long-range correlation structure of stride intervals is unaltered by significant peripheral sensory loss. This further supports the hypothesis that the central nervous system is involved in the regulation of long-range correlations. PMID- 17110520 TI - Respiratory impairment in a mouse model of amyotrophic lateral sclerosis. AB - Amyothrophic lateral sclerosis (ALS) is a progressive, lethal neuromuscular disease that is associated with the degeneration of cortical and spinal motoneurons, leading to atrophy of limb, axial, and respiratory muscles. Patients with ALS invariably develop respiratory muscle weakness and most die from pulmonary complications. Overexpression of superoxide dismutase 1 (SOD1) gene mutations in mice recapitulates several of the clinical and pathological characteristics of ALS and is therefore a valuable tool to study this disease. The present study is intended to evaluate an age-dependent progression of respiratory complications in SOD1(G93A) mutant mice. In each animal, baseline measurements of breathing pattern [i.e., breathing frequency and tidal volume (VT)], minute ventilation (VE), and metabolism (i.e., oxygen consumption and carbon dioxide production) were repeatedly sampled at variable time points between 10 and 20 wk of age with the use of whole-body plethysmographic chambers. To further characterize the neurodegeneration of breathing, VE was also measured during 5-min challenges of hypercapnia (5% CO(2)) and hypoxia (10% O(2)). At baseline, breathing characteristics and metabolism remained relatively unchanged from 10 to 14 wk of age. From 14 to 18 wk of age, there were significant (P < 0.05) increases in baseline VT, VE, and the ventilatory equivalent (VE/oxygen consumption). After 18 wk of age, there was a rapid decline in VE due to significant (P < 0.05) reductions in both breathing frequency and VT. Whereas little change in hypoxic VE responses occurred between 10 and 18 wk, hypercapnic VE responses were significantly (P < 0.05) elevated at 18 wk due to an augmented VT response. Like baseline breathing characteristics, hypercapnic VE responses also declined rapidly after 18 wk of age. The phenotypic profile of SOD1(G93A) mutant mice was apparently unique because similar changes in respiration and metabolism were not observed in SOD1 controls. The present results outline the magnitude and time course of respiratory complications in SOD1(G93A) mutant mice as the progression of disease occurs in this mouse model of ALS. PMID- 17110521 TI - Effects of neonatal maternal separation on neurochemical and sensory response to colonic distension in a rat model of irritable bowel syndrome. AB - Early life stress has been implicated as a risk factor for irritable bowel syndrome (IBS). We studied the effect of neonatal maternal separation on the visceromotor response and the expression of c-fos, 5-HT, and its receptors/transporters along the brain-gut axis in an animal model of IBS. Male neonatal Sprague-Dawley rats were randomly assigned to a 3-h daily maternal separation (MS) or nonhandling (NH) on postnatal days 2-21. Colorectal balloon distention (CRD) was performed for assessment of abdominal withdrawal reflex as a surrogate marker of visceral pain. Tissues from dorsal raphe nucleus in midbrain, lumbar-sacral cord, and distal colon were harvested for semiquantitative analysis of c-fos and 5-HT. The expression of 5-HT expression, 5-HT3 receptors, and 5-HT transporter were analyzed by RT-PCR. Pain threshold was significantly lower in MS than NH rats. The abdominal withdrawal reflex score in response to CRD in MS rats was significantly higher with distension pressures of 40, 60, and 80 mmHg. In MS rats, the number of c-fos-like immunoreactive nuclei at dorsal horn of lumbar sacral spinal cord increased significantly after CRD. 5-HT content in the spinal cord of MS rats was significant higher. In the colon, both 5-HT-positive cell number and 5-HT content were comparable between MS and NH groups before CRD. Post CRD only MS rats had significant increase in 5-HT content. Protein and mRNA expression levels of 5-HT3 receptors and 5-HT transporter were similar in MS and NH rats. Neonatal maternal separation stress predisposes rats to exaggerated neurochemical responses and visceral hyperalgesia in colon mimicking IBS. PMID- 17110522 TI - Defective hepatocyte aquaporin-8 expression and reduced canalicular membrane water permeability in estrogen-induced cholestasis. AB - Our previous work supports a role for aquaporin-8 (AQP8) water channels in rat hepatocyte bile formation mainly by facilitating the osmotically driven canalicular secretion of water. In this study, we tested whether a condition with compromised canalicular bile secretion, i.e., the estrogen-induced intrahepatic cholestasis, displays defective hepatocyte AQP8 functional expression. After 17alpha-ethinylestradiol administration (5 mg x kg body wt(-1).day(-1) for 5 days) to rats, the bile flow was reduced by 58% (P < 0.05). By subcellular fractionation and immunoblotting analysis, we found that 34 kDa AQP8 was significantly decreased by approximately 70% in plasma (canalicular) and intracellular (vesicular) liver membranes. However, 17alpha-ethinylestradiol induced cholestasis did not significantly affect the protein level or the subcellular localization of sinusoidal AQP9. Immunohistochemistry for liver AQPs confirmed these observations. Osmotic water permeability (P(f)) of canalicular membranes, measured by stopped-flow spectrophotometry, was significantly reduced (73 +/- 1 vs. 57 +/- 2 microm/s) in cholestasis, consistent with defective canalicular AQP8 functional expression. By Northern blotting, we found that AQP8 mRNA expression was increased by 115% in cholestasis, suggesting a posttranscriptional mechanism of protein level reduction. Accordingly, studies in primary cultured rat hepatocytes indicated that the lysosomal protease inhibitor leupeptin prevented the estrogen-induced AQP8 downregulation. In conclusion, hepatocyte AQP8 protein expression is downregulated in estrogen-induced intrahepatic cholestasis, presumably by lysosomal-mediated degradation. Reduced canalicular membrane AQP8 expression is associated with impaired osmotic membrane water permeability. Our data support the novel notion that a defective expression of canalicular AQP8 contributes as a mechanism for bile secretory dysfunction of cholestatic hepatocytes. PMID- 17110523 TI - Engineering novel VPAC2-selective agonists with improved stability and glucose lowering activity in vivo. AB - A previously described VPAC2-selective agonist, BAY 55-9837 (peptide HSDAVFTDNYTRLRKQVAAKKYLQSIKNKRY), had several limitations with respect to its potential as an insulin secretagogue for the treatment of type 2 diabetes. These limitations were primarily poor stability in aqueous buffer and short duration of action in vivo. In this report, we describe a series of novel analogs of BAY 55 9837 that were designed around the likely degradation mechanisms and structure activity relationship of this peptide with a view to overcoming its limitations. These analogs were tested for improved liquid stability and retention of VPAC2 selective binding and activation, as well as prolonged activity in vivo. Although several degradation mechanisms were possible based on the degradation pattern, it was determined that deamidation at the two asparagines (N9 and N28) was the major instability determinant. Changing these two asparagines to glutamines did not negatively affect VPAC2-selective binding and activation. The double glutamine mutein analog, BAY(Q9Q28), retained full VPAC2 activity and selectivity while displaying no significant degradation when stored at 40 degrees C for 4 weeks. This is in contrast to BAY 55-9837, which showed greater than 80% degradation when stored at 40 degrees C for 2 weeks. A cysteine was added to the C terminus of BAY(Q9Q28), followed by site-specific cysteine conjugation with a 22- or 43 kDa polyethylene glycol (PEG) to yield BAY(Q9Q28C32)PEG22 or BAY(Q9Q28C32)PEG43, respectively. These PEGylated peptides retain the ability to selectively bind and activate the VPAC2 receptor and have prolonged glucose-lowering activity in vivo. PMID- 17110524 TI - Ca2+-independent, inhibitory effects of cyclic adenosine 5'-monophosphate on Ca2+ regulation of phosphoinositide 3-kinase C2alpha, Rho, and myosin phosphatase in vascular smooth muscle. AB - We have recently demonstrated in vascular smooth muscle (VSM) that membrane depolarization by high KCl induces Ca(2+)-dependent Rho activation and myosin phosphatase (MLCP) inhibition (Ca(2+)-induced Ca(2+)-sensitization) through the mechanisms involving phosphorylation of myosin-targeting protein 1 (MYPT1) and 17 kDa protein kinase C (PKC)-potentiated inhibitory protein of PP1 (CPI-17). In the present study, we investigated whether and how cAMP affected Ca(2+)-dependent MLCP inhibition by examining the effects of forskolin, cell-permeable dibutyryl cAMP (dbcAMP), and isoproterenol. Forskolin, but not its inactive analog 1,9 dideoxyforskolin, inhibited KCl-induced contraction and the 20-kDa myosin light chain (MLC) phosphorylation without inhibiting Ca(2+) mobilization in rabbit aortic VSM. dbcAMP mimicked these forskolin effects. We recently suggested that Ca(2+)-mediated Rho activation is dependent on class II alpha-isoform of phosphoinositide 3-kinase (PI3K-C2alpha). Forskolin inhibited KCl-induced stimulation of PI3K-C2alpha activity. KCl-induced membrane depolarization stimulated Rho in a manner dependent on a PI3K but not PKC and stimulated phosphorylation of MYPT1 at Thr(850) and CPI-17 at Thr(38) in manners dependent on both PI3K and Rho kinase, but not PKC. Forskolin, dbcAMP, and isoproterenol inhibited KCl-induced Rho activation and phosphorylation of MYPT1 and CPI-17. Consistent with these data, forskolin, isoproterenol, a PI3K inhibitor, or a Rho kinase inhibitor, but not a PKC inhibitor, abolished KCl-induced diphosphorylation of MLC. These observations indicate that cAMP inhibits Ca(2+) mediated activation of the MLCP-regulating signaling pathway comprising PI3K C2alpha, Rho, and Rho kinase in a manner independent of Ca(2+) and point to the novel mechanism of the cAMP actions in the regulation of vascular smooth muscle contraction. PMID- 17110525 TI - Hippocampus, ageing, and taste memories. AB - Previous studies have shown that ageing may induce deficits in hippocampal dependent learning and memory tasks, the spatial task being most extensively applied in rats. It is proposed that taste learning and memory tasks may assist in understanding the ageing of memory systems, giving access to a more complete picture. Taste learning tasks allow us to explore a variety of learning phenomena in safe and aversive memories using similar behavioral procedures. In demanding the same sensory, response, and motivational requirements, this approach provides reliable comparisons between the performance of hippocampal lesioned and aged rats in different types of memory. Present knowledge on the effect of both ageing and hippocampal damage in complex taste learning phenomena is reviewed. Besides inducing deficits in hippocampal-dependent phenomena, such as blocking of conditioned taste aversion, while at the same time leaving intact nonhippocampal dependent effects, such as latent inhibition, ageing is also associated with an increased neophobia by previous aversive taste memories and enhanced taste aversion conditioning which cannot be explained by age-related changes in taste or visceral distress sensitivity. In all, the results indicate a peculiar organization of the memory systems during aging that cannot be explained by a general cognitive decline or exclusively by the decay of the hippocampal function. PMID- 17110526 TI - Brain regions responsible for the expression of conditioned taste aversion in rats. AB - Conditioned taste aversion (CTA) is acquired when the ingestion of a food is followed by malaise. CTA is a kind of fear learning making animals avoid subsequent intake of the food and show aversive behavior to the taste of the food. To elucidate the brain regions responsible for the expression of CTA, our previous electrophysiological and recent c-fos immunohistochemical studies have been reviewed. Among a variety of brain regions including the parabrachial nucleus, amygdala, insular cortex, supramammillary nucleus, nucleus accumbens, and ventral pallidum that are involved in different phases of CTA expression, the enhanced taste sensitivity to facilitate detection of the conditioned stimulus may originate in the central nucleus of the amygdala and the hedonic shift, from positive to negative, may originate in the basolateral nucleus of the amygdala. PMID- 17110527 TI - Molecular signaling during taste aversion learning. AB - Behavioral and neural assessment tools have been used to identify cellular and molecular events that occur during taste aversion acquisition. Studies described here include an assessment of taste information processing and taste-illness association using fos-like immunoreactivity (FLI) to mark populations of cells that react strongly to the taste conditioned stimulus (CS), the illness unconditioned stimulus (US), or the pairing of CS and US. Exposure to a novel, but not a familiar, CS taste (saccharin) was found to induce robust increases in FLI in some, but not all, brain regions previously implicated in taste processing or taste aversion learning. Striking effects of taste novelty on FLI were found in central amygdala (CNA) and insular cortex (IC) but not in basolateral amygdala (BLA), pontine parabrachial nucleus (PBN), or nucleus of the solitary tract (NTS). Of those regions responding to taste novelty, only CNA showed significant elevations in FLI in response to the US, LiCl. In additional studies, FLI was examined after an effective training experience, novel CS-US pairing, and compared with an ineffective one, familiar CS-US pairing. After CS-US pairing, taste novelty modulated FLI in virtually all the regions previously implicated in conditioned taste aversion (CTA) learning, including PBN, CNA, BLA, IC, as well as NTS. Thus, a distributed and interdependent neural CTA circuit is mapped using this method, and the use of localized lesion and inactivation studies promises to further define the functional role of structures within this circuit. PMID- 17110528 TI - Proceedings of the symposium on brain mechanisms of taste recognition memory and neural plasticity held at the ninth European congress of psychology on 6 July 2005. PMID- 17110529 TI - Serotonin-type 3 receptors mediate intestinal lipid-induced satiation and Fos like immunoreactivity in the dorsal hindbrain. AB - Several gastrointestinal stimuli, including some intestinal nutrients, have been shown to exert their satiating effect via activation of serotonin type-3 (5 HT(3)) receptors. The presence of lipids in the small intestine potently suppresses food intake; however, whether 5-HT(3) receptors play a role in this response has not been directly examined. Therefore, using the selective 5-HT(3) receptor antagonist ondansetron, we tested the hypothesis that duodenal infusion of lipid suppresses intake of both sucrose solution and chow through 5-HT(3) receptor activation. Rats duodenally infused with 72 and 130 mM Intralipid suppressed 1-h 15% sucrose intake by 33 and 67%, respectively. Suppression of sucrose intake by 72 mM Intralipid was significantly attenuated by ondansetron at all doses tested (0.5, 1.0, 2.0, and 5.0 mg/kg ip), whereas the lowest effective dose of ondansetron to attenuate suppression of intake by 130 mM Intralipid was 1.0 mg/kg. Furthermore, infusion of 130 mM Intralipid suppressed 1- and 4-h chow intake by 35 and 20%, respectively. Ondansetron administered as low as 0.5 mg/kg significantly attenuated 1-h Intralipid-induced suppression of chow intake and completely reversed the suppression by 4 h. Administration of ondansetron alone did not alter sucrose or chow intake compared with vehicle injection at any time. Finally, to test whether Intralipid-induced neuronal activation of the dorsal vagal complex is mediated by 5-HT(3) receptors, Fos-like immunoreactivity (Fos LI) was quantified in ondansetron-pretreated rats following intestinal lipid infusion. Ondansetron (1 mg/kg) significantly attenuated duodenal intralipid induced Fos-LI in the dorsal hindbrain. These data support the hypothesis that 5 HT(3) receptors mediate both satiation, as well as hindbrain neuronal responses evoked by intestinal lipids. PMID- 17110530 TI - Cardiac changes during arousals from non-REM sleep in healthy volunteers. AB - Our aim was to evaluate cardiac changes evoked by spontaneous and sound-induced arousals from sleep. Cardiac responses to spontaneous and auditory-induced arousals were recorded during overnight sleep studies in 28 young healthy subjects (14 males, 14 females) during non-rapid eye movement sleep. Computerized analysis was applied to assess beat-to-beat changes in heart rate, atrio ventricular conductance, and ventricular repolarization from 30 s before to 60 s after the auditory tone. During both types of arousals, the most consistent change was the increase in the heart rate (in 62% of spontaneous and in 89% of sound-induced arousals). This was accompanied by an increase or no change in PR interval and by a decrease or no change in QT interval. The magnitude of all cardiac changes was significantly higher for tone-induced vs. spontaneous arousals (mean +/- SD for heart rate: +9 +/- 8 vs. +13 +/- 9 beats per min; for PR prolongation: 14 +/- 16 vs. 24 +/- 22 ms; for QT shortening: -12 +/- 6 vs. -20 +/- 9 ms). The prevalence of transient tachycardia and PR prolongation was also significantly higher for tone-induced vs. spontaneous arousals (tachycardia: 85% vs. 57% of arousals, P < 0.001; PR prolongation: 51% vs. 25% of arousals, P < 0.001). All cardiac responses were short-lasting (10-15 s). We conclude that cardiac pacemaker region, conducting system, and ventricular myocardium may be under independent neural control. Prolongation of atrio-ventricular delay may serve to increase ventricular filling during arousal from sleep. Whether prolonged atrio-ventricular conductance associated with increased sympathetic outflow to the ventricular myocardium contributes to arrhythmogenesis during sudden arousal from sleep remains to be evaluated. PMID- 17110531 TI - Region-specific changes in the phosphorylation of ERK1/2 and ERK5 in rat micturition pathways following cyclophosphamide-induced cystitis. AB - Chronic inflammation of the urinary bladder generates hyperalgesia and allodynia. Growing evidence suggests a role of ERK in mediating somatic and visceral pain processing. In the present studies, we characterized and compared the activation of two ERK isoforms, ERK1/2 and ERK5, in micturition pathways, including the urinary bladder, lumbosacral dorsal root ganglia (DRG), and spinal cord in adult female and male rats before and after cyclophosphamide (CYP)-induced bladder inflammation. Results showed differential activation of ERK1/2 and ERK5 in these regions following cystitis. The level of phospho-ERK1/2 but not phospho-ERK5 was increased in the urinary bladder; the level of phospho-ERK5 but not phospho ERK1/2 was increased in DRG; and the level of phospho-ERK1/2 but not phospho-ERK5 was increased in lumbar spinal cord following cystitis compared with control. Cystitis-induced upregulation of phospho-ERK1/2 and phospho-ERK5 was time dependent and showed similar patterns in female and male rats. The level of phospho-ERK1/2 in bladder was increased at 2 and 8 h after CYP injection; the level of phospho-ERK5 in DRG was increased at 8 and 48 h after CYP injection; and the level of phospho-ERK1/2 in lumbar spinal cord was increased at 48 h after CYP injection. The result that phospho-ERK5 was exclusively increased in DRG neurons, while phospho-ERK1/2 was increased in the spinal cord and the urinary bladder after cystitis, suggests a region-specific effect of neurotrophins on micturition pathways following bladder inflammation. PMID- 17110532 TI - Rate of tension redevelopment is not modulated by sarcomere length in permeabilized human, murine, and porcine cardiomyocytes. AB - The increase in Ca(2+) sensitivity of isometric force development along with sarcomere length (SL) is considered as the basis of the Frank-Starling law of the heart, possibly involving the regulation of cross-bridge turnover kinetics. Therefore, the Ca(2+) dependencies of isometric force production and of the cross bridge-sensitive rate constant of force redevelopment (k(tr)) were determined at different SLs (1.9 and 2.3 mum) in isolated human, murine, and porcine permeabilized cardiomyocytes. k(tr) was also determined in the presence of 10 mM inorganic phosphate (P(i)), which interfered with the force-generating cross bridge transitions. The increases in Ca(2+) sensitivities of force with SL were very similar in human, murine, and porcine cardiomyocytes (DeltapCa(50): approximately 0.11). k(tr) was higher (P < 0.05) in mice than in humans or pigs at all Ca(2+) concentrations ([Ca(2+)]) [maximum k(tr) (k(tr,max)) at a SL of 1.9 mum and pCa 4.75: 1.33 +/- 0.11, 7.44 +/- 0.15, and 1.02 +/- 0.05 s(-1), in humans, mice, and pigs, respectively] but k(tr) did not depend on SL in any species. Moreover, when the k(tr) values for each species were expressed relative to their respective maxima, similar Ca(2+) dependencies were obtained. Ten millimolar P(i) decreased force to approximately 60-65% and left DeltapCa(50) unaltered in all three species. P(i) increased k(tr,max) by a factor of approximately 1.6 in humans and pigs and by a factor of approximately 3 in mice, independent of SL. In conclusion, species differences exert a major influence on k(tr), but SL does not appear to modulate the cross-bridge turnover rates in human, murine, and porcine hearts. PMID- 17110533 TI - Differential response of type 2 deiodinase gene expression to photoperiod between photoperiodic Fischer 344 and nonphotoperiodic Wistar rats. AB - The molecular basis of seasonal or nonseasonal breeding remains unknown. Although laboratory rats are generally regarded as photoperiod-insensitive species, the testicular weight of the Fischer 344 (F344) strain responds to photoperiod. Recently, it was clarified that photoperiodic regulation of type 2 iodothyronine deiodinase (Dio2) in the mediobasal hypothalamus (MBH) is critical in photoperiodic gonadal regulation. Strain-dependent differences in photoperiod sensitivity may now provide the opportunity to address the regulatory mechanism of seasonality by studying Dio2 expression. Therefore, in the present study, we examined the effect of photoperiod on Dio2 expression in photoperiod-sensitive F344 and photoperiod-insensitive Wistar rats. A statistically significant difference was observed between short and long days in terms of testicular weight and Dio2 expression in the F344 strain, while no difference was observed in the Wistar strain. These results suggest that differential responses of the Dio2 gene to photoperiod may determine the strain-dependent differences in photoperiod sensitivity in laboratory rats. PMID- 17110534 TI - Reduction in antioxidant defenses may contribute to ochratoxin A toxicity and carcinogenicity. AB - Ochratoxin A (OTA) is a renal carcinogen in rodents. Its human health significance is unclear. It likely depends upon the mechanism of carcinogenesis. In a previous microarray study a reduction in nuclear factor-erythroid 2 p45 related factor 2 (Nrf2)-dependent gene expression was observed in the kidney but not in the liver of rats fed OTA up to 12 months. Nrf2 regulates detoxification and antioxidant gene expression. The present report shows that OTA decreased the protein expression of several markers of the Nrf2-regulated gene battery in kidney in vivo indicating that the effects observed at mRNA level may be of biological significance. The OTA-mediated Nrf2 response could be reproduced in an NRK renal cell line and in primary hepatocyte cultures. In in vitro systems, an OTA-mediated inhibition of Nrf2 activity was demonstrated by electrophoretic mobility shift and Antioxidant Regulatory Element-driven luciferase reporter assays. The reduction of Nrf2-regulated gene expression resulted in oxidative DNA damage as evidenced by formation of abasic sites in vitro and confirmed in kidney in vivo. All OTA-mediated effects observed were prevented by pretreatment of cell cultures with inducers of Nrf2 activity. Our data suggest that reduction of cellular defense against oxidative stress by Nrf2 inhibition may be a plausible mechanism of OTA nephrotoxicity and carcinogenicity. PMID- 17110535 TI - A cytokinin perception mutant colonized by Rhizobium in the absence of nodule organogenesis. AB - In legumes, Nod-factor signaling by rhizobia initiates the development of the nitrogen-fixing nodule symbiosis, but the direct cell division stimulus that brings about nodule primordia inception in the root cortex remains obscure. We showed that Lotus japonicus plants homozygous for a mutation in the HYPERINFECTED 1 (HIT1) locus exhibit abundant infection-thread formation but fail to initiate timely cortical cell divisions in response to rhizobial signaling. We demonstrated that the corresponding gene encodes a cytokinin receptor that is required for the activation of the nodule inception regulator Nin and nodule organogenesis. PMID- 17110536 TI - The ground state of the pseudogap in cuprate superconductors. AB - We present studies of the electronic structure of La(2-x)BaxCuO4, a system where the superconductivity is strongly suppressed as static spin and charge orders or "stripes" develop near the doping level of x = (1/8). Using angle-resolved photoemission and scanning tunneling microscopy, we detect an energy gap at the Fermi surface with magnitude consistent with d-wave symmetry and with linear density of states, vanishing only at four nodal points, even when superconductivity disappears at x = (1/8). Thus, the nonsuperconducting, striped state at x = (1/8) is consistent with a phase-incoherent d-wave superconductor whose Cooper pairs form spin-charge-ordered structures instead of becoming superconducting. PMID- 17110537 TI - A gain-of-function mutation in a cytokinin receptor triggers spontaneous root nodule organogenesis. AB - Legume root nodules originate from differentiated cortical cells that reenter the cell cycle and form organ primordia. We show that perception of the phytohormone cytokinin is a key element in this switch. Mutation of a Lotus japonicus cytokinin receptor gene leads to spontaneous development of root nodules in the absence of rhizobia or rhizobial signal molecules. The mutant histidine kinase receptor has cytokinin-independent activity and activates an Escherichia coli two component phosphorelay system in vivo. Mutant analysis shows that cytokinin signaling is required for cell divisions that initiate nodule development and defines an autoregulated process where cytokinin induction of nodule stem cells is controlled by shoots. PMID- 17110538 TI - Don't grandfather coal plants. PMID- 17110539 TI - Election 2006. Science awaits impact of Democratic sweep in Congress. PMID- 17110541 TI - Election 2006. Gordon steps up to House science post. PMID- 17110540 TI - Election 2006. Environmentalists see a greener Congress. PMID- 17110542 TI - Election 2006. Stem cell supporters hail results, but political lessons aren't clear. PMID- 17110543 TI - Election 2006. Scientists get out the word. PMID- 17110544 TI - Election 2006. Elsewhere on the election front. PMID- 17110545 TI - Global climate change. False alarm: Atlantic conveyor belt hasn't slowed down after all. PMID- 17110546 TI - Chinese drug research. Novartis invests $100 million in Shanghai. PMID- 17110547 TI - Physics. Electronic nuisance changes its ways. PMID- 17110548 TI - Public health. SARS and bird flu veteran to take WHO helm. PMID- 17110549 TI - Paleogenetics. The dawn of Stone Age genomics. PMID- 17110550 TI - Paleogenetics. A Neandertal legacy? PMID- 17110551 TI - High-temperature superconductivity turns 20. High Tc: the mystery that defies solution. PMID- 17110552 TI - High-temperature superconductivity turns 20. The next big hurdle: economics. PMID- 17110553 TI - High-temperature superconductivity turns 20. Determined duo scored a victory for small-scale science. PMID- 17110554 TI - Why aren't there more scientists advocating for funding? PMID- 17110555 TI - Fighting waterborne infectious diseases. PMID- 17110556 TI - Debating the Worth of NCCAM Research. PMID- 17110557 TI - Research funding. NIH in the post-doubling era: realities and strategies. PMID- 17110558 TI - Psychology. Money is material. PMID- 17110559 TI - Geology. Why do freezing rocks break? PMID- 17110560 TI - Computer science. What do robots dream of? PMID- 17110561 TI - Cell biology. Sara splits the signal. PMID- 17110562 TI - Chemistry. Breaking the H2 marriage and reuniting the couple. PMID- 17110563 TI - Biochemistry. RNA polymerase, a scrunching machine. PMID- 17110565 TI - Composites in armor. AB - Composite materials are traditionally regarded as materials that can save energy in large structures associated with transport. They are used to produce lightweight structures for fuel-efficient aircraft such as the new Boeing 787 Dreamliner; lightweight cars from Lotus, Ferrari and TVR; and high-speed trains, speedboats, and racing yachts. Now, however, some of the most interesting applications of composites are those where the materials are used to save lives and protect property by absorbing the energy of projectiles, impacts, and crashes. PMID- 17110566 TI - In quest of virtual tests for structural composites. AB - The difficult challenge of simulating diffuse and complex fracture patterns in tough structural composites is at last beginning to yield to conceptual and computational advances in fracture modeling. Contributing successes include the refinement of cohesive models of fracture and the formulation of hybrid stress strain and traction-displacement models that combine continuum (spatially averaged) and discrete damage representations in a single calculation. Emerging hierarchical formulations add the potential of tracing the damage mechanisms down through all scales to the atomic. As the models near the fidelity required for their use as virtual experiments, opportunities arise for reducing the number of costly tests needed to certify safety and extending the design space to include material configurations that are too complex to certify by purely empirical methods. PMID- 17110567 TI - Nanoparticle polymer composites: where two small worlds meet. AB - The mixing of polymers and nanoparticles is opening pathways for engineering flexible composites that exhibit advantageous electrical, optical, or mechanical properties. Recent advances reveal routes to exploit both enthalpic and entropic interactions so as to direct the spatial distribution of nanoparticles and thereby control the macroscopic performance of the material. For example, by tailoring the particle coating and size, researchers have created self-healing materials for improved sustainability and self-corralling rods for photovoltaic applications. A challenge for future studies is to create hierarchically structured composites in which each sublayer contributes a distinct function to yield a mechanically integrated, multifunctional material. PMID- 17110568 TI - Rapid temporal reversal in predator-driven natural selection. AB - As the environment changes, will species be able to adapt? By conducting experiments in natural environments, biologists can study how evolutionary processes such as natural selection operate through time. We predicted that the introduction of a terrestrial predator would first select for longer-legged lizards, which are faster, but as the lizards shifted onto high twigs to avoid the predator, selection would reverse toward favoring the shorter-legged individuals better able to locomote there. Our experimental studies on 12 islets confirmed these predictions within a single generation, thus demonstrating the rapidity with which evolutionary forces can change during times of environmental flux. PMID- 17110570 TI - Resilient machines through continuous self-modeling. AB - Animals sustain the ability to operate after injury by creating qualitatively different compensatory behaviors. Although such robustness would be desirable in engineered systems, most machines fail in the face of unexpected damage. We describe a robot that can recover from such change autonomously, through continuous self-modeling. A four-legged machine uses actuation-sensation relationships to indirectly infer its own structure, and it then uses this self model to generate forward locomotion. When a leg part is removed, it adapts the self-models, leading to the generation of alternative gaits. This concept may help develop more robust machines and shed light on self-modeling in animals. PMID- 17110571 TI - Solid-state thermal rectifier. AB - We demonstrated nanoscale solid-state thermal rectification. High-thermal conductivity carbon and boron nitride nanotubes were mass-loaded externally and inhomogeneously with heavy molecules. The resulting nanoscale system yields asymmetric axial thermal conductance with greater heat flow in the direction of decreasing mass density. The effect cannot be explained by ordinary perturbative wave theories, and instead we suggest that solitons may be responsible for the phenomenon. Considering the important role of electrical rectifiers (diodes) in electronics, thermal rectifiers have substantial implications for diverse thermal management problems, ranging from nanoscale calorimeters to microelectronic processors to macroscopic refrigerators and energy-saving buildings. PMID- 17110572 TI - Reversible, metal-free hydrogen activation. AB - Although reversible covalent activation of molecular hydrogen (H2) is a common reaction at transition metal centers, it has proven elusive in compounds of the lighter elements. We report that the compound (C6H2Me3)2PH(C6F4)BH(C6F5)2 (Me, methyl), which we derived through an unusual reaction involving dimesitylphosphine substitution at a para carbon of tris(pentafluorophenyl) borane, cleanly loses H2 at temperatures above 100 degrees C. Preliminary kinetic studies reveal this process to be first order. Remarkably, the dehydrogenated product (C6H2Me3)2P(C6F4)B(C6F5)2 is stable and reacts with 1 atmosphere of H2 at 25 degrees C to reform the starting complex. Deuteration studies were also carried out to probe the mechanism. PMID- 17110569 TI - Sequencing and analysis of Neanderthal genomic DNA. AB - Our knowledge of Neanderthals is based on a limited number of remains and artifacts from which we must make inferences about their biology, behavior, and relationship to ourselves. Here, we describe the characterization of these extinct hominids from a new perspective, based on the development of a Neanderthal metagenomic library and its high-throughput sequencing and analysis. Several lines of evidence indicate that the 65,250 base pairs of hominid sequence so far identified in the library are of Neanderthal origin, the strongest being the ascertainment of sequence identities between Neanderthal and chimpanzee at sites where the human genomic sequence is different. These results enabled us to calculate the human-Neanderthal divergence time based on multiple randomly distributed autosomal loci. Our analyses suggest that on average the Neanderthal genomic sequence we obtained and the reference human genome sequence share a most recent common ancestor approximately 706,000 years ago, and that the human and Neanderthal ancestral populations split approximately 370,000 years ago, before the emergence of anatomically modern humans. Our finding that the Neanderthal and human genomes are at least 99.5% identical led us to develop and successfully implement a targeted method for recovering specific ancient DNA sequences from metagenomic libraries. This initial analysis of the Neanderthal genome advances our understanding of the evolutionary relationship of Homo sapiens and Homo neanderthalensis and signifies the dawn of Neanderthal genomics. PMID- 17110573 TI - Bedrock fracture by ice segregation in cold regions. AB - The volumetric expansion of freezing pore water is widely assumed to be a major cause of rock fracture in cold humid regions. Data from experiments simulating natural freezing regimes indicate that bedrock fracture results instead from ice segregation. Fracture depth and timing are also numerically simulated by coupling heat and mass transfer with a fracture model. The depth and geometry of fractures match those in Arctic permafrost and ice-age weathering profiles. This agreement supports a conceptual model in which ice segregation in near-surface permafrost leads progressively to rock fracture and heave, whereas permafrost degradation leads episodically to melt of segregated ice and rock settlement. PMID- 17110574 TI - The impact of boreal forest fire on climate warming. AB - We report measurements and analysis of a boreal forest fire, integrating the effects of greenhouse gases, aerosols, black carbon deposition on snow and sea ice, and postfire changes in surface albedo. The net effect of all agents was to increase radiative forcing during the first year (34 +/- 31 Watts per square meter of burned area), but to decrease radiative forcing when averaged over an 80 year fire cycle (-2.3 +/- 2.2 Watts per square meter) because multidecadal increases in surface albedo had a larger impact than fire-emitted greenhouse gases. This result implies that future increases in boreal fire may not accelerate climate warming. PMID- 17110575 TI - Solar wind neon from Genesis: implications for the lunar noble gas record. AB - Lunar soils have been thought to contain two solar noble gas components with distinct isotopic composition. One has been identified as implanted solar wind, the other as higher-energy solar particles. The latter was puzzling because its relative amounts were much too large compared with present-day fluxes, suggesting periodic, very high solar activity in the past. Here we show that the depth dependent isotopic composition of neon in a metallic glass exposed on NASA's Genesis mission agrees with the expected depth profile for solar wind neon with uniform isotopic composition. Our results strongly indicate that no extra high energy component is required and that the solar neon isotope composition of lunar samples can be explained as implantation-fractionated solar wind. PMID- 17110576 TI - Sara endosomes and the maintenance of Dpp signaling levels across mitosis. AB - During development, cells acquire positional information by reading the concentration of morphogens. In the developing fly wing, a gradient of the transforming growth factor-beta (TGF-beta)-type morphogen decapentaplegic (Dpp) is transduced into a gradient of concentration of the phosphorylated form of the R-Smad transcription factor Mad. The endosomal protein Sara (Smad anchor for receptor activation) recruits R-Smads for phosphorylation by the type I TGF-beta receptor. We found that Sara, Dpp, and its type I receptor Thickveins were targeted to a subpopulation of apical endosomes in the developing wing epithelial cells. During mitosis, the Sara endosomes and the receptors therein associated with the spindle machinery to segregate into the two daughter cells. Daughter cells thereby inherited equal amounts of signaling molecules and thus retained the Dpp signaling levels of the mother cell. PMID- 17110577 TI - Abortive initiation and productive initiation by RNA polymerase involve DNA scrunching. AB - Using single-molecule DNA nanomanipulation, we show that abortive initiation involves DNA "scrunching"--in which RNA polymerase (RNAP) remains stationary and unwinds and pulls downstream DNA into itself--and that scrunching requires RNA synthesis and depends on RNA length. We show further that promoter escape involves scrunching, and that scrunching occurs in most or all instances of promoter escape. Our results support the existence of an obligatory stressed intermediate, with approximately one turn of additional DNA unwinding, in escape and are consistent with the proposal that stress in this intermediate provides the driving force to break RNAP-promoter and RNAP-initiation-factor interactions in escape. PMID- 17110578 TI - Initial transcription by RNA polymerase proceeds through a DNA-scrunching mechanism. AB - Using fluorescence resonance energy transfer to monitor distances within single molecules of abortively initiating transcription initiation complexes, we show that initial transcription proceeds through a "scrunching" mechanism, in which RNA polymerase (RNAP) remains fixed on promoter DNA and pulls downstream DNA into itself and past its active center. We show further that putative alternative mechanisms for RNAP active-center translocation in initial transcription, involving "transient excursions" of RNAP relative to DNA or "inchworming" of RNAP relative to DNA, do not occur. The results support a model in which a stressed intermediate, with DNA-unwinding stress and DNA-compaction stress, is formed during initial transcription, and in which accumulated stress is used to drive breakage of interactions between RNAP and promoter DNA and between RNAP and initiation factors during promoter escape. PMID- 17110579 TI - N-linked glycosylation of folded proteins by the bacterial oligosaccharyltransferase. AB - N-linked protein glycosylation is found in all domains of life. In eukaryotes, it is the most abundant protein modification of secretory and membrane proteins, and the process is coupled to protein translocation and folding. We found that in bacteria, N-glycosylation can occur independently of the protein translocation machinery. In an in vitro assay, bacterial oligosaccharyltransferase glycosylated a folded endogenous substrate protein with high efficiency and folded bovine ribonuclease A with low efficiency. Unfolding the eukaryotic substrate greatly increased glycosylation. We propose that in the bacterial system, glycosylation sites are located in flexible parts of folded proteins, whereas the eukaryotic cotranslational glycosylation evolved to a mechanism presenting the substrate in a flexible form before folding. PMID- 17110580 TI - New strategies for the elimination of polio from India. AB - The feasibility of global polio eradication is being questioned as a result of continued transmission in a few localities that act as sources for outbreaks elsewhere. Perhaps the greatest challenge is in India, where transmission has persisted in Uttar Pradesh and Bihar despite high coverage with multiple doses of vaccine. We estimate key parameters governing the seasonal epidemics in these areas and show that high population density and poor sanitation cause persistence by not only facilitating transmission of poliovirus but also severely compromising the efficacy of the trivalent vaccine. We analyze strategies to counteract this and show that switching to monovalent vaccine may finally interrupt virus transmission. PMID- 17110581 TI - The psychological consequences of money. AB - Money has been said to change people's motivation (mainly for the better) and their behavior toward others (mainly for the worse). The results of nine experiments suggest that money brings about a self-sufficient orientation in which people prefer to be free of dependency and dependents. Reminders of money, relative to nonmoney reminders, led to reduced requests for help and reduced helpfulness toward others. Relative to participants primed with neutral concepts, participants primed with money preferred to play alone, work alone, and put more physical distance between themselves and a new acquaintance. PMID- 17110582 TI - Generation of gut-homing IgA-secreting B cells by intestinal dendritic cells. AB - Normal intestinal mucosa contains abundant immunoglobulin A (IgA)-secreting cells, which are generated from B cells in gut-associated lymphoid tissues (GALT). We show that dendritic cells (DC) from GALT induce T cell-independent expression of IgA and gut-homing receptors on B cells. GALT-DC-derived retinoic acid (RA) alone conferred gut tropism but could not promote IgA secretion. However, RA potently synergized with GALT-DC-derived interleukin-6 (IL-6) or IL-5 to induce IgA secretion. Consequently, mice deficient in the RA precursor vitamin A lacked IgA-secreting cells in the small intestine. Thus, GALT-DC shape mucosal immunity by modulating B cell migration and effector activity through synergistically acting mediators. PMID- 17110583 TI - NPAS1 regulates branching morphogenesis in embryonic lung. AB - Drosophila trachealess (Trl), master regulator of tracheogenesis, has no known functional mammalian homolog. We hypothesized that genes similar to trachealess regulate lung development. Quantitative (Q)RT-PCR and immunostaining were used to determine spatial and temporal patterns of npas1 gene expression in developing murine lung. Immunostaining for alpha-smooth muscle actin demonstrated myofibroblasts, and protein gene product (PGP)9.5 identified neuroendocrine cells. Branching morphogenesis of embryonic lung buds was analyzed in the presence of antisense or sense oligodeoxynucleotides (ODN). Microarray analyses were performed to screen for changes in gene expression in antisense-treated lungs. QRT-PCR was used to validate the altered expression of key genes identified on the microarrays. We demonstrate that npas1 is expressed in murine embryonic lung. npas1 mRNA peaks early at Embryonic Day (E)10.5-E11.5, then drops to low levels. Sequencing verifies the identity of npas1 transcripts in embryonic lung. NPAS1 immunostaining occurs in nuclei of parabronchial mesenchymal cells, especially at the tracheal bifurcation. Arnt, the murine homolog of Tango (the heterodimerization partner for Trl) is also expressed in developing lung but at constant levels. npas1- or arnt-antisense ODN inhibit lung branching morphogenesis, with altered myofibroblast development and increased pulmonary neuroendocrine cells. On microarrays, we identify > 50 known genes down-regulated by npas1-antisense, including multiple genes regulating cell migration and cell differentiation. QRT-PCR confirms significantly decreased expression of the neurogenic genes RBP-Jk and Tle, and three genes involved in muscle development: beta-ig-h3, claudin-11, and myocardin. Npas1 can regulate myofibroblast distribution, branching morphogenesis, and neuroendocrine cell differentiation in murine embryonic lung. PMID- 17110584 TI - Comparison of environmental and mutational variation in flowering time in Arabidopsis. AB - Developmental dynamics can be influenced by external and endogenous factors in a more or less analogous manner. To compare the phenotypic effects of (i) environmental [i.e. standard (stPhP) and extended (exPhP) photoperiods] changes in Arabidopsis wild types and (ii) endogenous genetic variation in eav1-eav61 early flowering mutants, two temporal indicators were analysed, the time to bolting (DtB) and the number of leaves (TLN). It was found that DtB and TLN are differentially affected in different environmental and genetic contexts, and some factors of dynamic convergence were identified. The quantitative response to photoperiod is markedly contingent on the phototrophic input for DtB, but less so for TLN. To discriminate the light quantity and period components in DtB, two novel temporal indicators were determined, LtB (photosynthetic time to bolting) and PChron (DtB h(-1) of photoperiod), respectively. The use of PChron results in a coincidence of the variation profiles across stPhP and exPhP, interpreted as a buffering of the trophic response. Unlike natural accessions and later flowering mutants, the variation profiles across stPhP and eav mutants are significantly divergent, pointing to differences in environmental and genetic variation in flowering time. Yet, phenocopy effects and dynamic convergence between wild-type and mutant profiles are detected by using exPhP and the LtB indicator. Additional analyses of the cauline leaf number (CLN) show that the apical and basal boundaries of the primary inflorescence vary co-ordinately. The finding that the correlativity between CLN and TLN changes across photoperiods suggests that different states of intra-connectedness are involved in ontogenetic specification of flowering time and embodied in the primary inflorescence. PMID- 17110585 TI - Loss of ovule identity induced by overexpression of the fertilization-related kinase 2 (ScFRK2), a MAPKKK from Solanum chacoense. AB - In order to gain information about protein kinases acting during plant fertilization and embryogenesis, a reverse genetic approach was used to determine the role of protein kinases expressed in reproductive tissues. Two cDNA clones named ScFRK1 and ScFRK2 (Solanum chacoense fertilization-related kinase 1 and 2) were isolated from an expressed sequence tag (EST) library normalized for weakly expressed genes in fertilized ovaries. These showed significant sequence similarities to members of the mitogen-activated protein kinase kinase kinase (MAPKKK) family. RNA gel blot and RNA in situ hybridization analyses confirmed the strong up-regulation of ScFRK2 in ovules after fertilization. In addition, ScFRK2 mRNAs accumulate during early ovule development in the megasporocyte and in the integument of developing ovules. Overexpression of ScFRK2 led to the production of fruits with a severely reduced number of seeds. The seeds that were produced also exhibited developmental retardation. Analysis of ovaries prior to fertilization showed that the seedless phenotype was caused by a homeotic conversion of ovules into carpel-like structures. The present observations are consistent with the role of ScFRK2 in pre- and post-fertilization events. Furthermore, overexpression of ScFRK2 led to changes in the expression of the class D floral homeotic gene ScFBP11, suggesting that the ScFRK2 kinase may interact, directly or indirectly, with the FBP7/11 pathway that directs establishment of ovule identity. PMID- 17110586 TI - 3-D cell-level chlorophyll fluorescence imaging of ozone-injured sunflower leaves using a new passive light microscope system. AB - A passive light microscope system has been developed, capable of reconstructing an extended-focus 3-D cell-level image of chlorophyll fluorescence and Phi(PSII) of intact attached leaves using a limited number of focal plane images of chlorophyll fluorescence. Using this system, the relationships between the depth of the mesophyll cells in spongy tissue and the intensity of the chlorophyll fluorescence and the Phi(PSII) were investigated in sunflower leaves exposed to 300 ppb ozone for 12 h at a PPFD of 300 micromol m(-2) s(-1) actinic light. After ozone exposure, fluorescence intensity (F) largely decreased in the cells just under the epidermal cells (within approximately 20 microm of the epidermal cells), but the sites where fluorescence intensity decreased had no relationship to the position of the stomata. By contrast, the distribution of Phi(PSII) showed no change after the ozone exposure. These findings suggest that ozone-induced inhibition occurs in the cells just under the epidermal cells by reducing the light absorption of the chloroplasts, while the operating quantum efficiency of PSII photochemistry is maintained. PMID- 17110587 TI - Barley transcript profiles under dehydration shock and drought stress treatments: a comparative analysis. AB - A microarray including 1654 cDNAs, mainly derived from dehydration-shocked barley leaf tissues, was utilized to monitor expression changes in leaves of barley plants subjected to slow drying conditions (7 d and 11 d: 7d-WS and 11d-WS) in soil and after rehydration. The results were compared with those obtained under shock-like conditions imposed with a 6 h dehydration treatment. A total number of 173 transcripts (approximately 10% of all transcripts profiled) were declared up- or down-regulated in at least one of the conditions tested. The majority of the transcripts were regulated by only one of the drought treatments, with 57% of the differentially expressed transcripts exclusively affected in the dehydration shock treatment, 6% at 7d-WS, 14% at 11d-WS, and 6% after rehydration. Irrespective of the low percentage of transcripts (10%) with similar expression changes between shock and slow stress treatments, a sizeable portion of these transcripts shared a common expression trend under the different drought treatment conditions, as evidenced by low but significant correlations between the fast occurring and the 7d-WS and 11d-WS treatments (r=0.32 and 0.41, P=0.001, respectively). These results are discussed with respect to the merit of different dehydration treatments in the investigation of the changes in transcript profiling. PMID- 17110588 TI - Calcium pectate chemistry controls growth rate of Chara corallina. AB - Pectin, a normal constituent of cell walls, caused growth rates to accelerate to the rates in living cells when supplied externally to isolated cell walls of Chara corallina. Because this activity was not reported previously, the activity was investigated. Turgor pressure (P) was maintained in isolated walls or living cells using a pressure probe in culture medium. Pectin from various sources was supplied to the medium. Ca and Mg were the dominant inorganic elements in the wall. EGTA or pectin in the culture medium extracted moderate amounts of wall Ca and essentially all the wall Mg, and wall growth accelerated. Removing the external EGTA or pectin and replacing with fresh medium returned growth to the original rate. A high concentration of Ca2+ quenched the accelerating activity of EGTA or pectin and caused gelling of the pectin, physically inhibiting wall growth. Low pH had little effect. After the Mg had been removed, Ca-pectate in the wall bore the longitudinal load imposed by P. Removal of this Ca caused the wall to burst. Live cells and isolated walls reacted similarly. It was concluded that Ca cross-links between neighbouring pectin molecules were strong wall bonds that controlled wall growth rates. The central role of Ca-pectate chemistry was illustrated by removing Ca cross-links with new pectin (wall "loosening"), replacing vacated cross-links with new Ca2+ ("Ca2+-tightening"), or adding new cross-links with new Ca-pectate that gelled ("gel tightening"). These findings establish a molecular model for growth that includes wall deposition and assembly for sustained growth activity. PMID- 17110589 TI - Vacuolar transporters and their essential role in plant metabolism. AB - Following the unequivocal demonstration that plants contain at least two types of vacuoles, scientists studying this organelle have realized that the plant 'vacuome' is far more complex than they expected. Some fully developed cells contain at least two large vacuoles, with different functions. Remarkably, even a single vacuole may be subdivided and fulfil several functions, which are supported in part by the vacuolar membrane transport systems. Recent studies, including proteomic analyses for several plant species, have revealed the tonoplast transporters and their involvement in the nitrogen storage, salinity tolerance, heavy metal homeostasis, calcium signalling, guard cell movements, and the cellular pH homeostasis. It is clear that vacuolar transporters are an integrated part of a complex cellular network that enables a plant to react properly to changing environmental conditions, to save nutrients and energy in times of plenty, and to maintain optimal metabolic conditions in the cytosol. An overview is given of the main features of the transporters present in the tonoplast of plant cells in terms of their function, regulation, and relationships with the microheterogeneity of the vacuome. PMID- 17110590 TI - Randomized phase II three-arm trial with three platinum-based doublets in metastatic non-small-cell lung cancer. An Italian Trials in Medical Oncology study. AB - BACKGROUND: Many patients with advanced non-small-cell lung cancer (NSCLC) do not tolerate cisplatin-based regimens because of its nonhemathological toxicity. PATIENTS AND METHODS: We evaluated the response rate safety of new platinum analogue regimens, randomizing 147 patients with nonoperable IIIB/IV NSCLC to (i) carboplatin (area under the curve = 5 mg min/ml) on day 1 plus gemcitabine (GEM) (1000 mg/m(2)) on days 1 and 8 for six cycles; (ii) same regimen for three cycles followed by docetaxel (Taxotere) (40 mg/m(2)) on days 1 and 8 plus GEM (1250 mg/m(2)) on days 1 and 8 for three cycles; (iii) oxaliplatin (130 mg/m(2)) on day 1 plus GEM (1250 mg/m(2)) on days 1 and 8 for six cycles. RESULTS: Intention-to treat objective response rates were 25%, 25% and 30.6% in arms A, B and C, respectively. Median survival was 11.9, 9.2 and 11.3 months in arms A, B and C, respectively. Grade 3/4 neutropenia/anemia occurred in 29%/12.5%, 10%/16.5% and 8%/6% of arms A, B and C, respectively; grade 3/4 thrombocytopenia in 20.5%, 16.5% and 6%; grade 1/2 neurological toxicity in 43% of arm C. CONCLUSIONS: Oxaliplatin/GEM (arm C) had similar activity to carboplatin/GEM (arm A), but milder hematological toxicity and may be worth testing in a phase III study against carboplatin/GEM in patients not suitable for cisplatin. The sequential regimen gave no additional benefit. PMID- 17110591 TI - Temporal dynamics of plastic changes in human primary somatosensory cortex after finger webbing. AB - The primary somatosensory cortex (SI) exhibits a detailed topographic organization of the hand and fingers, which has been found to undergo plastic changes following modifications of the sensory input. Although the spatial properties of these changes have been extensively investigated, little is known about their temporal dynamics. In this study, we adapted the paradigm of finger webbing, in which 4 fingers are temporarily webbed together, hence modifying their sensory feedback. We used magnetoencephalography, to measure changes in the hand representation in SI, before, during, and after finger webbing for about 5 h. Our results showed a decrease in the Euclidean distance (ED) between cortical sources activated by electrical stimuli to the index and small finger 30 min after webbing, followed by an increase lasting for about 2 h after webbing, which was followed by a return toward baseline values. These results provide a unique frame in which the different representational changes occur, merging previous findings that were only apparently controversial, in which either increases or decreases in ED were reported after sensory manipulation for relatively long or short duration, respectively. Moreover, these observations further confirm that the mechanisms that underlie cortical reorganization are extremely rapid in their expression and, for the first time, show how brain reorganization occurs over time. PMID- 17110592 TI - Differential contributions of prefrontal, medial temporal, and sensory-perceptual regions to true and false memory formation. AB - The neural correlates of true memory formation (TMF) and false memory formation (FMF) were investigated using functional magnetic resonance imaging (fMRI). Using a parametric subsequent memory paradigm, encoding activity was analyzed as a function of whether it predicted subsequent hits to targets (TMF activity) or subsequent false alarms to critical lures (FMF activity). The fMRI analyses yielded 3 main findings. First, the left prefrontal cortex (PFC) was involved in both TMF and FMF activities. This finding is consistent with the evidence that semantic elaboration, which has been associated with left PFC, tends to enhance both true and false remembering. Second, the left posterior medial temporal lobes (MTLs) contributed to TMF but not to FMF activity. This finding is consistent with the notion that MTL is involved in the storage of a consciously, but not unconsciously, processed event. Third, late visual regions were engaged in both TMF and FMF activities, whereas early visual areas were involved primarily in TMF activity. This dissociation indicates that elaborative perceptual processing, but not basic sensory processing, contributes to false remembering. Taken together, the results suggest that FMF is an unintended consequence, or by-product, of elaborative semantic and visual encoding processes. PMID- 17110593 TI - Molecular coupling of a Ca2+-activated K+ channel to L-type Ca2+ channels via alpha-actinin2. AB - Cytoskeletal proteins are known to sculpt the structural architecture of cells. However, their role as bridges linking the functional crosstalk of different ion channels is unknown. Here, we demonstrate that a small conductance Ca(2+) activated K(+) channels (SK2 channel), present in a variety of cells, where they integrate changes in intracellular Ca(2+) concentration [Ca(2+)(i)] with changes in K(+) conductance and membrane potential, associate with L-type Ca(2+) channels; Ca(v)1.3 and Ca(v)1.2 through a physical bridge, alpha-actinin2 in cardiac myocytes. SK2 channels do not physically interact with L-type Ca(2+) channels, instead, the 2 channels colocalize via their interaction with alpha actinin2 cytoskeletal protein. The association of SK2 channel with alpha-actinin2 localizes the channel to the entry of external Ca(2+) source, which regulate the channel function. Furthermore, we demonstrated that the functions of SK2 channels in atrial myocytes are critically dependent on the normal expression of Ca(v)1.3 Ca(2+) channels. Null deletion of Ca(v)1.3 channel results in abnormal function of SK2 channel and prolongation of repolarization and atrial arrhythmias. Our study provides insight into the molecular mechanisms of the coupling of SK2 channel with voltage-gated Ca(2+) channel, and represents the first report linking the coupling of 2 different types of ion channels via cytoskeletal proteins. PMID- 17110594 TI - Hypoxia-induced endothelial proliferation requires both mTORC1 and mTORC2. AB - A central regulator of cell growth that has been implicated in responses to stress such as hypoxia is mTOR (mammalian Target Of Rapamycin). We have shown previously that mTOR is required for angiogenesis in vitro and endothelial cell proliferation in response to hypoxia. Here we have investigated mTOR-associated signaling components under hypoxia and their effects on cell proliferation in rat aortic endothelial cells (RAECs). Hypoxia (1% O(2)) rapidly (>30 minutes) and in a concentration-dependent manner promoted rapamycin-sensitive and sustained phosphorylation of mTOR-Ser2448 followed by nuclear translocation in RAECs. Similarly, hypoxia induced phosphorylation of the mTORC2 substrate Akt-Ser473 (3 to 6 hours at 1% O(2)) and a brief phosphorylation peak of the mTORC1 substrate S6 kinase-Thr389 (10 to 60 minutes). Phosphorylation of Akt was inhibited by mTOR knockdown and partially with rapamycin. mTOR knockdown, rapamycin, or Akt inhibition specifically and significantly inhibited proliferation of serum starved RAECs under hypoxia (P<0.05; n> or =4). Similarly, hypoxia induced Akt dependent and rapamycin-sensitive proliferation in mouse embryonic fibroblasts. This response was partially blunted by hypoxia-inducible factor-1alpha knockdown and not affected by TSC2 knockout. Finally, mTORC2 inhibition by rictor silencing, especially (P<0.001; n=7), and mTORC1 inhibition by raptor silencing, partially (P<0.05; n=7), inhibited hypoxia-induced RAEC proliferation. Thus, mTOR mediates an early response to hypoxia via mTORC1 followed by mTORC2, promoting endothelial proliferation mainly via Akt signaling. mTORC1 and especially mTORC2 might therefore play important roles in diseases associated with hypoxia and altered angiogenesis. PMID- 17110595 TI - A nuclear receptor corepressor-dependent pathway mediates suppression of cytokine induced C-reactive protein gene expression by liver X receptor. AB - C-reactive protein (CRP), the prototypical human acute phase protein, is an independent risk predictor of future cardiovascular events, both in healthy individuals and in patients with known cardiovascular disease. In addition, previous studies indicate that CRP might have direct proatherogenic properties. Ligand activation of the liver X receptor (LXR), a member of the nuclear hormone receptor superfamily, inhibits inflammatory gene expression in macrophages and attenuates the development of atherosclerosis in various animal models. We demonstrate herein that 2 synthetic LXR ligands, T0901317 and GW3965, inhibit interleukin-1beta/interleukin-6-induced CRP mRNA and protein expression in human hepatocytes. Knockdown of LXRalpha/beta by short interfering RNAs completely abolished the inhibitory effect of the LXR agonist T0901317 on cytokine-induced CRP gene transcription. Transient transfection experiments with 5'-deletion CRP promoter constructs identified a region from -125 to -256 relative to the initiation site that mediated the inhibitory effect of LXR ligands on CRP gene transcription. Depletion of the nuclear receptor corepressor by specific short interfering RNA increased cytokine-inducible CRP mRNA expression and promoter activity and reversed LXR ligand-mediated repression of CRP gene transcription. Chromatin immunoprecipitation assays indicated that nuclear receptor corepressor is present on the endogenous CRP promoter under basal conditions. Cytokine induced clearance of nuclear receptor corepressor complexes was inhibited by LXR ligand treatment, maintaining the CRP gene in a repressed state. Finally, treatment of C57Bl6/J mice with LXR ligands attenuated lipopolysaccharide-induced mouse CRP and serum amyloid P component gene expression in the liver, whereas no effect was observed in LXRalphabeta knockout mice. Our observations identify a novel mechanism of inflammatory gene regulation by LXR ligands. Thus, inhibition of CRP expression by LXR agonists may provide a promising approach to impact initiation and progression of atherosclerosis. PMID- 17110596 TI - Alpha1-adrenergic receptors activate AKT via a Pyk2/PDK-1 pathway that is tonically inhibited by novel protein kinase C isoforms in cardiomyocytes. AB - AKT is a potent antiapoptotic kinase, but its role in the cardioprotective actions of alpha(1)-adrenergic receptors (ARs) remains uncertain, because alpha(1)-ARs typically induce little-to-no AKT activation in most cardiomyocyte models. This study identifies a prominent alpha(1)-AR-dependent AKT activation pathway that is under tonic inhibitory control by novel protein kinase Cs (nPKCs) in neonatal rat cardiomyocyte cultures. We also implicate Pyk2, Pyk2 complex formation with PDK-1 and paxillin, and increased PDK-1-Y373/376 phosphorylation as the mechanism that links alpha(1)-AR activation to increased AKT phosphorylation. nPKCs (which are prominent alpha(1)-AR effectors) interfere with this alpha(1)-AR-dependent AKT activation by blocking Pyk2/PDK-1/paxillin complex formation and PDK-1-Y373/376 phosphorylation. Additional studies used an adenoviral-mediated overexpression strategy to show that Pyk2 exerts dual controls on antiapoptotic PDK-1/AKT and proapoptotic c-Jun N-terminal kinase (JNK) pathways. Although the high nPKC activity of most cardiomyocyte models favors Pyk2 signaling to JNK (and cardiac apoptosis), the cardioprotective actions of Pyk2 through the PDK-1/AKT pathway are exposed when PKC or JNK activation is prevented. Collectively, these studies identify JNK and AKT as functionally distinct downstream components of the alpha(1)-AR/Pyk2 signaling pathway. We also implicate nPKCs as molecular switches that control the balance of signaling via proapoptotic JNK and antiapoptotic PDK-1/AKT pathways, exposing a novel mechanism for nPKC-dependent regulation of cardiac hypertrophy and failure. PMID- 17110597 TI - Increasing ryanodine receptor open probability alone does not produce arrhythmogenic calcium waves: threshold sarcoplasmic reticulum calcium content is required. AB - Diastolic waves of Ca(2+) release have been shown to activate delayed afterdepolarizations as well as some cardiac arrhythmias. The aim of this study was to investigate whether increasing ryanodine receptor open probability alone or in the presence of beta-adrenergic stimulation produces diastolic Ca release from the sarcoplasmic reticulum (SR). When voltage-clamped rat ventricular myocytes were exposed to caffeine (0.5 to 1.0 mmol), diastolic Ca(2+) release was seen to accompany the first few stimuli but was never observed in the steady state. We attribute the initial phase of diastolic Ca(2+) release to a decrease in the threshold SR Ca(2+) content required to activate Ca(2+) waves and its subsequent disappearance to a decrease of SR content below this threshold. Application of isoproterenol (1 micromol/L) increased the amplitude of the systolic Ca(2+) transient and also the SR Ca(2+) content but did not usually produce diastolic Ca(2+) release. Subsequent addition of caffeine, however, resulted in diastolic Ca(2+) release. We estimated the time course of recovery of SR Ca(2+) content following recovery from emptying with a high (10 mmol/L) concentration of caffeine. Diastolic Ca(2+) release recommenced only when SR content had increased back to its final level. We conclude that increasing ryanodine receptor open probability alone does not produce arrhythmogenic diastolic Ca(2+) release because of the accompanying decrease of SR Ca(2+) content. beta-Adrenergic stimulation increases SR content and thereby allows the increased ryanodine receptor open probability to produce diastolic Ca(2+) release. The implications of these results for arrhythmias associated with abnormal ryanodine receptors are discussed. PMID- 17110598 TI - Induction of heat shock response protects the heart against atrial fibrillation. AB - There is evidence suggesting that heat shock proteins (HSPs) may protect against clinical atrial fibrillation (AF). We evaluated the effect of HSP induction in an in vitro atrial cell line (HL-1) model of tachycardia remodeling and in tachypaced isolated canine atrial cardiomyocytes. We also evaluated the effect of HSP induction on in vivo AF promotion by atrial tachycardia-induced remodeling in dogs. Tachypacing (3 Hz) significantly and progressively reduced Ca(2+) transients and cell shortening of HL-1 myocytes over 4 hours. These reductions were prevented by HSP-inducing pretreatments: mild heat shock, geranylgeranylacetone (GGA), and transfection with human HSP27 or the phosphorylation-mimicking HSP27-DDD. However, treatment with HSP70 or the phosphorylation-deficient mutant HSP27-AAA failed to alter tachycardia-induced Ca(2+) transient and cell-shortening reductions, and downregulation (short interfering RNA) of HSP27 prevented GGA-mediated protection. Tachypacing (3 Hz) for 24 hours in vitro significantly reduced L-type Ca(2+) current and action potential duration in canine atrial cardiomyocytes; these effects were prevented when tachypacing was performed in cells exposed to GGA. In vivo treatment with GGA increased HSP expression and suppressed refractoriness abbreviation and AF promotion in dogs subjected to 1-week atrial tachycardia-induced remodeling. In conclusion, our findings indicate that (1) HSP induction protects against atrial tachycardia-induced remodeling, (2) the protective effect in HL-1 myocytes requires HSP27 induction and phosphorylation, and (3) the orally administered HSP inducer GGA protects against AF in a clinically relevant animal model. These findings advance our understanding of the biochemical determinants of AF and suggest the possibility that HSP induction may be an interesting novel approach to preventing clinical AF. PMID- 17110599 TI - Regulation of thromboxane receptor trafficking through the prostacyclin receptor in vascular smooth muscle cells: role of receptor heterodimerization. AB - BACKGROUND: Prostacyclin (PGI2) and thromboxane (TxA2) effect disparate outcomes for atherogenesis and the response to vascular injury; PGI2, a vasodilator and inhibitor of platelet aggregation, limits the deleterious actions of TxA2, a vasoconstrictor and platelet activator. Dimerization of their G protein-coupled receptors, IP and TP, evokes a modified cellular response through which IP/TP counter-balance may be effected. We examined the consequence of IP/TP interaction for the regulatory pathways of both receptors. METHODS AND RESULTS: TPalpha overexpressed in HEK293 cells or expressed endogenously in aortic smooth muscle cells (ASMCs) was internalized after selective activation of either TP or IP. Homologous trafficking of TP was unaltered by coexpression of IP. Heterologous sequestration of TPalpha required the physical presence of activated IP, in transfected and native cells, but was independent of IP signaling to adenylyl cyclase. Reciprocal heterologous regulation of IP, via activated TP, was evident in both HEK293 cells and ASMCs. Homologous TP internalization led to receptor retention and degradation. In contrast, when internalization was IP-induced, TPalpha was recycled to the cell surface in coexpressing HEK293 cells, but not in ASMCs, in accord with the postendocytotic pathway of IP. CONCLUSIONS: IP/TPalpha interaction permits reciprocal regulation of receptor endocytosis via the trafficking pathway determined by the activated dimeric partner. PMID- 17110600 TI - Lysophosphatidic acid induces thrombogenic activity through phosphatidylserine exposure and procoagulant microvesicle generation in human erythrocytes. AB - OBJECTIVE: Although erythrocytes have been suggested to play a role in blood clotting, mediated through phosphatidylserine (PS) exposure and/or PS-bearing microvesicle generation, an endogenous substance that triggers the membrane alterations leading to a procoagulant activity in erythrocytes has not been reported. We now demonstrated that lysophosphatidic acid (LPA), an important lipid mediator in various pathophysiological processes, induces PS exposure and procoagulant microvesicle generation in erythrocytes, which represent a biological significance resulting in induction of thrombogenic activity. METHODS AND RESULTS: In human erythrocytes, LPA treatment resulted in PS exposure on remnant cells and PS-bearing microvesicle generation in a concentration-dependent manner. Consistent with the microvesicle generation, scanning electron microscopic study revealed that LPA treatment induced surface changes, alteration of normal discocytic shape into echinocytes followed by spherocytes. Surprisingly, chelation of intracellular calcium did not affect LPA-induced PS exposure and microvesicle generation. On the other hand, protein kinase C (PKC) inhibitors significantly reduced PS exposure and microvesicle generation induced by LPA, reflecting the role of calcium-independent PKC. Activation of PKC was confirmed by Western blot analysis showing translocation of calcium-independent isoform, PKCzeta, to erythrocyte membrane. The activity of flippase, which is important in the maintenance of membrane asymmetry, was also inhibited by LPA. Furthermore, LPA-exposed erythrocytes actually potentiated the thrombin generation as determined by prothrombinase assay and accelerated the coagulation process initiated by recombinant human tissue factor in plasma. The adherence of erythrocytes to endothelial cells, another important feature of thrombogenic process, was also stimulated by LPA treatment. CONCLUSIONS: These results suggested that LPA-exposed erythrocytes could make an important contribution to thrombosis mediated through PS exposure and procoagulant microvesicle generation. PMID- 17110601 TI - Vascular endothelial growth factor receptor 2 plays a role in the activation of aortic endothelial cells by oxidized phospholipids. AB - OBJECTIVE: Previous studies have shown that oxidized products of PAPC (Ox-PAPC) regulate cell transcription of interleukin-8, LDL receptor, and tissue factor. This upregulation takes place in part through the activation of sterol regulatory element-binding protein (SREBP) and Erk 1/2. The present studies identify vascular endothelial growth factor receptor 2 (VEGFR2) as a major regulator in the activation of SREBP and Erk 1/2 in endothelial cells activated by Ox-PAPC. METHODS AND RESULTS: Ox-PAPC induced the phosphorylation of VEGFR2 at Tyr1175 in human aortic endothelial cells. Inhibitors and siRNA for VEGFR2 decreased the transcription of interleukin-8, LDL receptor, and tissue factor in response to Ox PAPC and the activation of SREBP and Erk 1/2, which mediate this transcription. We provide evidence that the activation of VEGFR2 is rapid, sustained, and c-Src dependent. CONCLUSIONS: These data point to a major role of VEGFR2 in endothelial regulation by oxidized phospholipids which accumulate in atherosclerotic lesions and apoptotic cells. PMID- 17110602 TI - Angiopoietin-like protein3 regulates plasma HDL cholesterol through suppression of endothelial lipase. AB - OBJECTIVE: A low level of high-density lipoprotein (HDL) in plasma has been recognized as an aspect of metabolic syndrome and as a crucial risk factor of cardiovascular events. However, the physiological regulation of plasma HDL levels has not been completely defined. Current studies aim to reveal the contribution of angiopoietin-like protein3 (angptl3), previously known as a plasma suppressor of lipoprotein lipase, to HDL metabolism. METHODS AND RESULTS: Angptl3-deficient mice showed low plasma HDL cholesterol and HDL phospholipid (PL), and which were increased by ANGPTL3 supplementation via adenovirus. In vitro, ANGPTL3 inhibited the phospholipase activity of endothelial lipase (EL), which hydrolyzes HDL-PL and hence decreases plasma HDL levels, through a putative heparin-binding site in the N-terminal domain of ANGPTL3. Post-heparin plasma in Angptl3-knockout mice had higher phospholipase activity than did that in wild-type mice, suggesting that the activity of endogenous EL is elevated in Angptl3-deficient mice. Furthermore, we established an ELISA system for human ANGPTL3 and found that plasma ANGPTL3 levels significantly correlated with plasma HDL cholesterol and HDL-PL levels in human subjects. CONCLUSIONS: Angptl3 acts as an inhibitor of EL and may be involved in the regulation of plasma HDL cholesterol and HDL-PL levels in humans and rodents. PMID- 17110604 TI - Triglyceride:high-density lipoprotein cholesterol effects in healthy subjects administered a peroxisome proliferator activated receptor delta agonist. AB - OBJECTIVE: Exercise increases fatty acid oxidation (FAO), improves serum high density lipoprotein cholesterol (HDLc) and triglycerides (TG), and upregulates skeletal muscle peroxisome proliferator activated receptor (PPAR)delta expression. In parallel, PPARdelta agonist-upregulated FAO would induce fatty acid uptake (via peripheral lipolysis), and influence HDLc and TG-rich lipoprotein particle metabolism, as suggested in preclinical models. METHODS AND RESULTS: Healthy volunteers were allocated placebo (n=6) or PPARdelta agonist (GW501516) at 2.5 mg (n=9) or 10 mg (n=9), orally, once-daily for 2 weeks while hospitalized and sedentary. Standard lipid/lipoproteins were measured and in vivo fat feeding studies were conducted. Human skeletal muscle cells were treated with GW501516 in vitro and evaluated for lipid-related gene expression and FAO. Serum TG trended downwards (P=0.08, 10 mg), whereas TG clearance post fat-feeding improved with drug (P=0.02). HDLc was enhanced in both treatment groups (2.5 mg P=0.004, 10 mg P<0.001) when compared with the decrease in the placebo group ( 11.5+/-1.6%, P=0.002). These findings complimented in vitro cell culture results whereby GW501516 induced FAO and upregulated CPT1 and CD36 expression, in addition to a 2-fold increase in ABCA1 (P=0.002). However, LpL expression remained unchanged. CONCLUSIONS: This is the first report of a PPARdelta agonist administered to man. In this small study, GW501516 significantly influenced HDLc and TGs in healthy volunteers. Enhanced in vivo serum fat clearance, and the first demonstrated in vitro upregulation in human skeletal muscle fat utilization and ABCA1 expression, suggests peripheral fat utilization and lipidation as potential mechanisms toward these HDL:TG effects. PMID- 17110603 TI - GPVI potentiation of platelet activation by thrombin and adhesion molecules independent of Src kinases and Syk. AB - OBJECTIVE: The present study investigates the role of Src and Syk tyrosine kinases in signaling by G-protein coupled and platelet adhesion receptors. METHODS AND RESULTS: Using Syk-/- platelets or the Src kinase inhibitor PP2, we demonstrate a critical role for Src and Syk kinases in mediating lamellipodia formation on VWF, collagen, CRP, fibrinogen, and fibronectin. In all cases, the spreading defect was overcome by addition of thrombin. Conversely, platelet aggregation and alphaIIb beta3 activation induced by thrombin was similar to controls, arguing against a functional role for Src and Syk in alphaIIb beta3 activation. Unexpectedly, CRP potentiated integrin alphaIIb beta3 activation and platelet aggregation induced by subthreshold concentrations of thrombin in Syk-/- platelets or in the presence of the Src kinase inhibitor PP2. Potentiation in the presence of PP2 was lost in the absence of FcRgamma-chain or GPVI confirming that it was mediated through the immunoglobulin receptor. Further delineation of this PP2-resistant synergy revealed that PAR4 could trigger the enhanced response in combination with CRP. CONCLUSIONS: We show that Syk is critical for lamellipodia formation on a range of immobilized proteins but that this can be overcome by addition of thrombin. Further, we reveal a novel role for GPVI in supporting thrombin-induced activation, independent of Syk and Src kinases. PMID- 17110605 TI - Association between a leukotriene C4 synthase gene promoter polymorphism and coronary artery calcium in young women: the Muscatine Study. AB - OBJECTIVE: A majority of the recognized risk factors for atherosclerosis and the development of cardiovascular disease have been derived from the study of older populations who have already manifested clinical symptoms. If risk factors can be identified earlier in life, such as genetic variation, preventive measures may be taken before overt symptoms of pathology have manifested, and when treatments may be most effective. METHODS AND RESULTS: In an effort to identify individuals at increased risk for cardiovascular disease, we genotyped 732 members of the Muscatine Study Longitudinal Adult Cohort for candidate genetic markers associated with several pathogenetic processes. We identified age-adjusted increased risks for coronary artery calcium (OR 4.29; 95% CI 1.78, 10.31) and increased mean carotid artery intimal-medial thickness associated with the ( 444)A>C promoter polymorphism of Leukotriene C4 Synthase (LTC4S) in women. There were no similar associations in men. CONCLUSIONS: LTC4S plays a key role in the process of inflammation as the rate limiting enzyme in the conversion of arachidonic acid to cysteinyl-leukotrienes, important mediators of inflammatory responses. The (-444)C variant upregulates LTC4S mRNA expression, increasing the synthesis of proinflammatory leukotrienes. Our results support genetic variation modifying inflammatory pathways as an important mechanism in the development of atherosclerosis. PMID- 17110606 TI - Origin of neointimal smooth muscle: we've come full circle. PMID- 17110607 TI - Regulation of TLR4 expression is a tale about tail. PMID- 17110608 TI - Coupling eNOS uncoupling to the innate immune response. PMID- 17110609 TI - Evaluation of differences in coronary plaque mechanical behavior in individuals with and without type 2 diabetes mellitus. PMID- 17110611 TI - Early recanalization after intravenous administration of recombinant tissue plasminogen activator as assessed by pre- and post-thrombolytic angiography in acute ischemic stroke patients. AB - BACKGROUND AND PURPOSE: Recanalization rates after the intravenous (IV) recombinant tissue plasminogen activator (rt-PA) treatment have been poorly studied in acute stroke. METHODS: CT angiography was performed before IV rt-PA in all patients and digital subtraction angiography was undertaken for intra arterial thrombolysis in cases of no improvement after rt-PA infusion. RESULTS: Forty-five patients were treated with IV rt-PA. Initial CT angiography showed relevant arterial occlusions in 35 patients. Recanalization after rt-PA therapy was demonstrated by digital subtraction angiography in 7 of the 31 patients with the occlusion on initial CT angiography: 2/16 in the internal carotid or proximal middle cerebral artery, 3/11 in the distal middle cerebral artery and 2/4 in the basilar artery occlusion. CONCLUSIONS: The early recanalization rate after IV rt PA use was very low in cases with large proximal arterial occlusions. CT angiography before IV rt-PA may be useful for the prediction of its efficacy. PMID- 17110613 TI - Role of antiplatelets in carotid artery stenting. PMID- 17110615 TI - Predisposition to carotid atherosclerosis in ICARAS dental substudy. PMID- 17110616 TI - Erythropoietin receptor expression in non-small cell lung carcinoma: a question of antibody specificity. AB - Immunohistochemical studies on formalin-fixed, paraffin-embedded (FFPE) tissue utilizing polyclonal antibodies form the cornerstone of many reports claiming to demonstrate erythropoietin receptor (EPOR) expression in malignant tissue. Recently, Elliott et al. (Blood 2006;107:1892-1895) reported that the antibodies commonly used to detect EPOR expression also detect non-EPOR proteins, and that their binding to EPOR was severely abrogated by two synthetic peptides based on the sequence of heat shock protein (HSP) 70, HSP70-2, and HSP70-5. We have investigated the specificity of the C20 antibody for detecting EPOR expression in non-small cell lung carcinoma (NSCLC) utilizing tissue microarrays. A total of 34 cases were available for study. Antibody absorbed with peptide resulted in marked suppression of cytoplasmic staining compared with nonabsorbed antibody. Four tumors that initially showed a membranous pattern of staining retained this pattern with absorbed antibody. Positive membranous immunoreactivity was also observed in 6 of 30 tumors that originally showed a predominantly cytoplasmic pattern of staining. Using the C20 antibody for Western blots, we detected three main bands, at 100, 66, and 59 kDa. Preincubation with either peptide caused abolition of the 66-kDa band, which contains non-EPOR sequences including heat shock peptides. These results call into question the significance of previous immunohistochemical studies of EPOR expression in malignancy and emphasize the need for more specific anti-EPOR antibodies to define the true extent of EPOR expression in neoplastic tissue. PMID- 17110617 TI - No contribution of multipotent mesenchymal stromal cells to liver regeneration in a rat model of prolonged hepatic injury. AB - Multipotent mesenchymal stromal (MS) cells from adult bone marrow are a cell population that can be expanded to large numbers in culture. MS cells might be differentiated toward hepatocytes in vitro and thus are promising candidates for therapeutic applications in vivo. The efficacy of bone marrow-derived MS cells versus hepatocytes to contribute to liver regeneration was compared in a rat model of prolonged toxic hepatic injury. Liver damage was induced by injection of carbon tetrachloride (CCl(4)) or allyl alcohol (AA) with and without retrorsine (R) pretreatment. MS cells or hepatocytes of wild-type F344 rats were injected into dipeptidyl peptidase IV (DPPIV)-deficient syngeneic rats. Hepatocyte chimerism was higher after intraportal hepatocyte transplantation in the R/AA group (mean maximal cluster size [MCS] = 21 cells) compared with the R/CCl(4) treatment group (MCS = 18). No hepatocyte engraftment was outlined following post transplant CCl(4) injection only, whereas mere AA injection resulted in small clusters of donor-derived hepatocytes (MCS = 2). Intraparenchymal injection of hepatocytes was associated with a MCS = 11 after R/AA treatment and a MCS = 6 after AA administration alone. Redistribution of MS cells to the liver was shown after intraportal and intraparenchymal injection. In contrast to hepatocyte transplantation, however, donor-derived DPPIV-positive cells could not be demonstrated in any recipient after MS cell transplantation. Data from the present study indicate that a well-defined population of MS cells obtained according to established standard protocols does not differentiate into hepatocytes in vivo when transplanted under regenerative conditions, in which the application of hepatocytes results in stable hepatic engraftment. PMID- 17110618 TI - Human mesenchymal stromal cells regulate initial self-renewing divisions of hematopoietic progenitor cells by a beta1-integrin-dependent mechanism. AB - In previous reports, we have demonstrated that only direct cell-cell contact with stromal cells, such as the murine stromal cell line AFT024, was able to alter the cell division kinetics and self-renewing capacity of hematopoietic progenitor cells (HPC). Because beta(1)-integrins were shown to be crucial for the interaction of HPC with the bone marrow microenvironment, we have studied the role of beta(1)-integrins in the regulation of self-renewing cell divisions. For this purpose, we used primary human mesenchymal stromal (MS) cells as in vitro surrogate niche and monitored the division history and subsequent functional fate of individually plated CD34(+)133(+) cells in the absence or presence of an anti beta(1)-integrin blocking antibody by time-lapse microscopy and subsequent long term culture-initiating cell (LTC-IC) assays. beta(1)-Integrin-mediated contact with MS cells significantly increased the proportion of asymmetrically dividing cells and led to a substantial increase of LTC-IC. Provided that beta(1)-integrin mediated contact was available within the first 72 hours, human MS cells were able to recruit HPC into cell cycle and accelerate their division kinetics without loss of stem cell function. Activation of beta(1)-integrins by ligands alone (e.g., fibronectin and vascular cell adhesion molecule-1) was not sufficient to alter the cell division symmetry and promote self-renewal of HPC, thus indicating an indirect effect. These results have provided evidence that primary human MS cells are able to induce self-renewing divisions of HPC by a beta(1)-integrin-dependent mechanism. PMID- 17110619 TI - Identification of human oral keratinocyte stem/progenitor cells by neurotrophin receptor p75 and the role of neurotrophin/p75 signaling. AB - This study was undertaken to determine whether human oral keratinocyte stem cells characteristically express higher levels of the low-affinity neurotrophin receptor p75 and to elucidate the function of p75 in oral keratinocytes. Examination of their expression patterns and cell-cycling status in vivo showed that p75 was exclusively expressed in the basal cell layer of both the tips of the papillae and the deep rete ridges. These immunostaining patterns suggest a cluster organization; most p75(+) cells did not actively cycle in vivo. Cell sorting showed that cells in the p75(+) subset were smaller and possessed higher in vitro proliferative capacity and clonal growth potential than the p75(-) subset. Clonal analysis revealed that holoclone-type (stem cell compartment), meroclone-type (intermediate compartment), and paraclone-type (transient amplifying cell compartment) cells, previously identified in skin and the ocular surface, were present in human oral mucosal epithelium. Holoclone-type cells showed stronger p75 expression at both the mRNA and protein level than did meroclone- and paraclone-type cells. Among the several neurotrophins, nerve growth factor (NGF) and neurotrophin-3 stimulated p75(+) oral keratinocyte cell proliferation, and only NGF protected them from apoptosis. Our in vivo and in vitro findings indicate that p75 is a potential marker of oral keratinocyte stem/progenitor cells and that some neurotrophin/p75 signaling affects cell growth and survival. PMID- 17110620 TI - Differentiation of adult mouse olfactory precursor cells into hair cells in vitro. AB - Many forms of deafness result from degeneration of the sensory cells for hearing, the hair cells in the cochlea. Stem cells offer a potential cell-based therapy for the treatment of deafness. Here, we investigate whether adult olfactory precursor cells can differentiate into hair cells in culture. Precursor cells were isolated from mouse olfactory neuroepithelium, were sphere-forming, showed proliferative capacity, and contained cells expressing neuronal and non-neuronal proteins. To induce differentiation, precursor cells were cocultured with cochlear cells and/or cochlear supernatant. Differentiated precursor cells were immunopositive for specific hair cell markers, including myosin VIIa, FM1-43, calretinin, phalloidin, and espin, and resembled hair cells anatomically and immunocytochemically in culture. The results demonstrate for the first time that adult olfactory precursor cells can differentiate into hair cell-like cells, thus providing a potential autotransplantation therapy for hearing loss. PMID- 17110621 TI - Embryonic stem cell-derived neurons form functional networks in vitro. AB - Embryonic stem (ES) cells provide a flexible and unlimited source for a variety of neuronal types. Because mature neurons establish neuronal networks very easily, we tested whether ES-derived neurons are capable of generating functional networks and whether these networks, generated in vitro, are capable of processing information. Single-cell electrophysiology with pharmacological antagonists demonstrated the presence of both excitatory and inhibitory synaptic connections. Extracellular recording with planar multielectrode arrays showed that spontaneous bursts of electrical activity are present in ES-derived networks with properties remarkably similar to those of hippocampal neurons. When stimulated with extracellular electrodes, ES-derived neurons fired action potentials, and the evoked electrical activity spread throughout the culture. A statistical analysis indicated that ES-derived networks discriminated between stimuli of different intensity at a single trial level, a key feature for an efficient information processing. Thus, ES-derived neurons provide a novel in vitro strategy to create functional networks with defined computational properties. PMID- 17110622 TI - Electrical stimulation modulates fate determination of differentiating embryonic stem cells. AB - A clear understanding of cell fate regulation during differentiation is key in successfully using stem cells for therapeutic applications. Here, we report that mild electrical stimulation strongly influences embryonic stem cells to assume a neuronal fate. Although the resulting neuronal cells showed no sign of specific terminal differentiation in culture, they showed potential to differentiate into various types of neurons in vivo, and, in adult mice, contributed to the injured spinal cord as neuronal cells. Induction of calcium ion influx is significant in this differentiation system. This phenomenon opens up possibilities for understanding novel mechanisms underlying cellular differentiation and early development, and, perhaps more importantly, suggests possibilities for treatments in medical contexts. PMID- 17110623 TI - Lapatinib: current status and future directions in breast cancer. AB - Lapatinib is an oral receptor tyrosine kinase inhibitor, targeting both the ErbB 1 and ErbB-2 receptors. Pre-clinical in vitro and in vivo models indicate that lapatinib is active as monotherapy, synergistically in combination with trastuzumab, and in trastuzumab-resistant cell lines. Early clinical trials also provide evidence in patients that lapatinib is active against breast cancer. This paper reviews results of phase II and III clinical trials of lapatinib in metastatic breast cancer, evidence for its potential in patients with brain metastases, and current clinical trials as adjuvant treatment in early-stage disease. Our improved understanding of the biology of breast cancer and the use of biomarkers for identification of specific subtypes is allowing us to bring patient-specific novel therapies such as lapatinib to the clinic. PMID- 17110624 TI - Appraising adjuvant aromatase inhibitor therapy. AB - Tamoxifen, once the gold standard adjuvant endocrine therapy for early breast cancer, is being challenged by third-generation aromatase inhibitors (AIs) that have demonstrated improved disease-free survival in a variety of adjuvant settings for early breast cancer. Tamoxifen and AIs have different safety profiles, which should allow physicians to begin to individualize treatment based on a patient's comorbidities and risk factors. Because of its properties as a partial estrogen agonist, tamoxifen has a positive effect on serum lipids and may confer a cardioprotective benefit, as well as a beneficial effect on bone health. However, tamoxifen increases the risk for endometrial cancer and cerebrovascular/thromboembolic events. In comparison, the major side effect of AIs is increased bone loss, which may heighten the risk for osteoporotic fractures and bone pain. Because of their superior efficacy and manageable side effects, AIs are a cost-effective alternative to tamoxifen, and clinical guidelines now embrace AIs as appropriate adjuvant therapy for hormone-sensitive early breast cancer. The anticipated results of ongoing trials will provide further insights into the long-term safety and application of AI therapy in the adjuvant setting. PMID- 17110625 TI - Nasal septum perforation in a bevacizumab-treated patient with metastatic breast cancer. PMID- 17110626 TI - Chronomodulated irinotecan, oxaliplatin, and leucovorin-modulated 5-Fluorouracil as ambulatory salvage therapy in patients with irinotecan- and oxaliplatin resistant metastatic colorectal cancer. AB - PURPOSE: To evaluate the activity and tolerability of salvage chronomodulated chemotherapy combining irinotecan (I), 5-fluorouracil/leucovorin (5-FU/LV), and oxaliplatin (O) (chronoIFLO) in patients with metastatic colorectal cancer (MCRC) and prior progression on four drugs. PATIENTS AND METHODS: Seventy-seven nonhospitalized MCRC patients received chronoIFLO every 3 weeks, with day 1: I (180 mg/m2 over 6 hours, with peak infusion rate at 05:00) and days 2-5: 5-FU/LV (700/300 mg/m2 per day over 12 hours, with peak flow rate at 04:00), and O (20 mg/m2 per day over 12 hours, with peak flow rate at 16:00). Toxicity and response were assessed every 3 weeks and every 2 months, respectively. RESULTS. Three or more prior chemotherapy lines were given to 75% of the patients. Two or more organs had metastatic disease in 65% of the patients. A median number of six courses of chronoIFLO was given. The main grade 3-4 toxicities were diarrhea (39% of the patients, 9% of the courses) and neutropenia (30% of the patients and 7% of the courses). Grade 3 peripheral sensory neuropathy occurred in 14% of the patients. Two patients achieved a partial response and 61 had stable disease, resulting in disease control for 82% of the patients. The median time to progression (TTP) was 5.5 months (95% confidence interval, 3.7-6.0). The median overall survival time was 14.2 months (9.8-17.3). Baseline performance status, serum carcinoembryonic antigen (CEA) level, and CEA doubling time were independent prognostic factors of TTP. CONCLUSIONS: ChronoIFLO safely and durably halted tumor progression in most extensively pretreated MCRC patients. PMID- 17110627 TI - Neoadjuvant use of hormonal therapy in elderly patients with early or locally advanced hormone receptor-positive breast cancer. AB - The management of the elderly patient with breast cancer is a challenge to the breast care team for a number of reasons. The higher rate of comorbidity in elderly patients increases the risk for complications and mortality following surgery and other adjuvant treatments such as chemotherapy and radiotherapy. The advent of using endocrine therapy in the neoadjuvant setting allows disease control and downstaging of tumors to allow less extensive surgery, with less morbidity compared with other available treatments. Tamoxifen has traditionally been the hormone therapy of choice for patients unable to undergo surgery, but development of resistance is a common feature. Newer third-generation aromatase inhibitors, in particular letrozole, are superior to tamoxifen in this setting with greater downstaging of tumor and disease control. The aromatase inhibitors are now the treatment of choice in elderly patients with estrogen receptor positive breast cancer who are being considered for neoadjuvant therapy. These drugs are particularly suitable to the needs of an elderly population. PMID- 17110628 TI - Treatment issues in clear cell carcinoma of the ovary: a different entity? AB - BACKGROUND: Ovarian clear cell carcinoma (OCCC) is a distinct histopathologic subtype of epithelial ovarian cancer (EOC) with an incidence of <5% of all ovarian malignancies. Our goal was to review the clinical features and management of patients with OCCC. METHODS AND RESULTS: We performed a PubMed search using the phrase "clear cell ovarian cancer." We reviewed 54 articles referring to OCCC. OCCC patients have a high incidence of stage I disease and frequently present with a large pelvic mass. Recurrences are more frequent with this entity than with other types of EOC. The clinical management of advanced EOC includes maximal cytoreduction and platinum plus paclitaxel-based chemotherapy. The survival rates of patients with advanced OCCC are lower than those of patients with advanced serous EOC (serous subtype). The poor response rate to platinum based regimens may be related to the intrinsic chemoresistance of these tumors. Despite their aggressive clinical course, OCCCs are still treated similarly to the other EOCs at the present time, because the rarity of these tumors prevents the conduction of randomized studies. CONCLUSION: Novel treatment approaches should be adopted in OCCC. Molecular-targeted therapies and effective new agents without cross-resistance to platinum compounds should be evaluated in a prospective clinical trial in OCCC. PMID- 17110629 TI - Chemotherapy given near the end of life by community oncologists for advanced non small cell lung cancer. AB - PURPOSE: To characterize the chemotherapy given near the end of life to advanced non-small cell lung cancer (NSCLC) patients treated in the community oncology setting using a medical records database. METHODS: We conducted a retrospective chart review of expired advanced (stage IIIb/IV) NSCLC patients treated with chemotherapy. Patients who initiated chemotherapy in 2000-2003 were eligible. Patient demographics, all chemotherapy including dose and schedule, and disease related events were collected. RESULTS: We report data from 10 community practices including 417 patients treated for advanced NSCLC in 2000-2003. The mean age was 67 years (median, 62 years) and 54% were male. Forty percent of patients were >69 years of age and 35% had an Eastern Cooperative Oncology Group performance status score of > or =2. First-line chemotherapy included combination therapy in 84% of patients. Second-line therapy was given to 56% of patients. Twenty-six percent of patients received third-line therapy, while 10% received fourth-line therapy and 5% received fifth-line therapy or greater. Patients received a mean of 6.1 cycles of chemotherapy. For patients receiving chemotherapy at the time of death, the mean line of therapy being given was second line. Chemotherapy was given within 1 month and 2 weeks of death to 43% and 20% of patients, respectively. CONCLUSION: The availability of new chemotherapeutic agents has caused a subsequent increase in the length of time patients are receiving chemotherapy with advanced NSCLC. This would suggest an increased use of chemotherapy near the end of life, which was identified in this study. PMID- 17110630 TI - Non-Hodgkin's lymphoma of mucosa-associated lymphoid tissue. AB - The concept of mucosa-associated lymphoid tissue (MALT) lymphomas was introduced by Isaacson and Wright [Cancer 1983; 52:1410-1416] in 1983. After more than 20 years of clinical research MALT lymphomas are now recognized as a distinct subtype of non-Hodgkin's lymphoma (NHL) with unique pathogenic, histological, and clinical features. Although this subtype of NHL occurs frequently, optimal management remains elusive. This manuscript reviews features of the clinical presentation, diagnosis, pathology, molecular characteristics, and management of both gastric and non-gastric MALT lymphoma. PMID- 17110631 TI - Radiation recall dermatitis with cefotetan: a case study. AB - Radiation recall dermatitis (RRD) is an inflammatory skin reaction that occurs in a previously irradiated body part following drug administration. This phenomenon may occur from days to years following exposure to ionizing radiation. The case of a 54-year-old Caucasian woman who was initially treated with external-beam radiation to the right thoracic region following the diagnosis of a poorly differentiated squamous cell carcinoma of the right lung is reported. She received four cycles of consolidated chemotherapy with docetaxel and carboplatin. Four months later, she was admitted to the hospital for acute cholecystitis and was placed on cefotetan. She developed a tender, erythematous rash on the posterior region of her right thorax 48 hours later. The drug was withdrawn, supportive care was instituted, and the patient subsequently improved. RRD should be suspected in patients who develop an erythematous rash in a previously irradiated region. To our knowledge this entity has not been associated with cefotetan previously. PMID- 17110632 TI - Bone loss and fracture risk associated with cancer therapy. AB - BACKGROUND: Cancer patients experience osteoporosis resulting from accelerated loss of bone mineral density (BMD) caused by their treatment. Such bone loss greatly increases the risk for fracture and can have other serious effects on quality of life. METHODS: In the current report, the author focuses on studies of cancer therapy-associated bone loss, its prevalence and pathogenesis, and resulting clinical impact. Options for management and prevention are also reviewed, including treatment guidelines where available. RESULTS: A variety of cancer therapies, including hormonal therapy, chemotherapy, and glucocorticoids, affect gonadal hormone production, which increases bone resorption and decreases BMD. Such bone loss occurs more rapidly and to a greater degree than normal age related osteoporosis, increases the risk for fracture and other morbidities, and decreases survival. Regular BMD screening and early intervention can prevent further decline in bone density and bone quality. Pharmacologic therapy with oral and i.v. bisphosphonates has been shown to slow bone loss in patients receiving cancer therapy, and the i.v. bisphosphonate zoledronic acid can increase BMD in patients with cancer treatment-related bone loss. Lifestyle changes, including supplementation with calcium and vitamin D, diet, and proper exercise, can also slow the rate of bone loss. CONCLUSIONS: Bone loss associated with various cancer therapies significantly affects bone health. Early initiation of bisphosphonates, when indicated, and lifestyle modification can improve patient outcomes. Education of patients and health care professionals regarding the importance of this complication and effective treatment options is essential. PMID- 17110633 TI - Benign breast diseases: classification, diagnosis, and management. PMID- 17110636 TI - Association between circulating white blood cell count and long-term incidence of age-related macular degeneration: the Blue Mountains Eye Study. AB - Inflammatory processes are implicated in the development and progression of age related macular degeneration (AMD). However, there are limited data on longitudinal associations between systemic markers of inflammation and AMD. The authors examined the prospective relation between the circulating white blood cell (WBC) count and early and late AMD in a population-based cohort of 3,654 participants, aged 49-97 years, in the Blue Mountains region, Australia. The main outcome of interest was the 10-year incidence of early and late AMD among individuals free from corresponding disease at the baseline (1992-1994). An elevated baseline WBC count was associated with early AMD incidence, independent of smoking and other major confounders. The multivariable relative risk comparing tertile 3 of WBC count (>6.7 x 10(9) cells/liter) with tertile 1 (/=5 cups/week versus 0-4 cups/week was 0.68 (95% confidence interval (CI): 0.43, 1.07). The authors observed a nonsignificant trend toward lower risk with higher consumption levels. Compared with that for 0-4 cups/week, the relative risks of MI were 0.84 (95% CI: 0.51, 1.38) for 5-7 cups/week, 0.65 (95% CI: 0.41, 1.03) for 2-3 cups/day, 0.64 (95% CI: 0.39, 1.04) for 4-5 cups/day, and 0.65 (95% CI: 0.37, 1.12) for >/=6 cups/day (p-trend = 0.07). Contrary to previous case-control studies, the authors concluded that coffee consumption does not increase MI risk. Coffee consumption of >/=5 cups/week was nonsignificantly inversely associated with MI risk among older Swedish women. PMID- 17110638 TI - Body size, dairy consumption, puberty, and risk of testicular germ cell tumors. AB - The etiology of testicular germ cell tumors (TGCTs) is poorly understood, with cryptorchidism and family history being the only well-established risk factors. Body size, age at puberty, and dairy consumption, however, have been suggested to be related to TGCTs. To clarify the relation of these variables to TGCT risk and to one another, the authors analyzed data from 767 cases and 928 controls enrolled in the Servicemen's Testicular Tumor Environmental and Endocrine Determinants Study (2002-2005). Overall, increased height was significantly related to risk (odds ratio (OR) = 1.83, 95% confidence interval (CI): 1.36, 2.45), though body mass index was not (OR = 1.06, 95% CI: 0.66, 1.69). There was no association with age at puberty, based on ages at first shaving (OR = 1.29, 95% CI: 0.96, 1.73), voice changing (OR = 0.97, 95% CI: 0.71, 1.32), and nocturnal emissions (OR = 1.00, 95% CI: 0.73, 1.37). Similarly, there was no relation with dairy consumption at any age between birth and 12th grade. These results suggest that height is a risk factor for TGCTs, but the relation is unlikely explained by childhood dairy consumption. As adult height is largely determined in the first 2 years of life, increased attention to events in this interval may help elucidate the etiology of TGCTs. PMID- 17110639 TI - Variants in estrogen biosynthesis genes, sex steroid hormone levels, and endometrial cancer: a HuGE review. AB - Variants in genes involved in estrogen biosynthesis are likely to be important in the etiology of endometrial cancer. This review summarizes data on variants in seven genes in the estrogen biosynthesis pathway and their relation to circulating levels of sex steroid hormones in women and to risk of endometrial cancer. Little or no association was found between genotypes of the cytochrome P 450 genes CYP11A1 (-528[TTTTA]n) or CYP17A1 (-34T/C) or the 17beta-hydroxysteroid dehydrogenase 1 gene HSD17B1 (Ser312Gly) and levels of progesterone, androgens, or estrogens. The position -34T/C variant in CYP17A1 appears to be associated with reduced risk of endometrial cancer, with those homozygous for the variant allele having about half the risk of those homozygous for the wild type. Linked variants in CYP19A1 (intron 4 [TTTA]n, intron 4 [TCT] insertion/deletion, exon 10 C/T) are related to some hormone levels and, based on two studies, to risk of endometrial cancer. For other genes (HSD3B1, HSD3B2, HSD17B2), no information is available on these associations. Results indicate the need to study other variants and haplotypes in these genes, particularly CYP17A1 and CYP19A1, as well as variants in other genes involved in hormone biosynthesis and metabolism pathways. Larger studies or combined studies that allow for investigation of gene gene and gene-environment interactions are warranted. PMID- 17110640 TI - Receiver operating characteristic curve inference from a sample with a limit of detection. AB - The receiver operating characteristic curve is a commonly used tool for evaluating biomarker usefulness in clinical diagnosis of disease. Frequently, biomarkers being assessed have immeasurable or unreportable samples below some limit of detection. Ignoring observations below the limit of detection leads to negatively biased estimates of the area under the curve. Several correction methods are suggested in the areas of mean estimation and testing but nothing regarding the receiver operating characteristic curve or its summary measures. In this paper, the authors show that replacement values below the limit of detection, including those suggested, result in the same biased area under the curve when properly accounted for, but they also provide guidance on the usefulness of these values in limited situations. The authors demonstrate maximum likelihood techniques leading to asymptotically unbiased estimators of the area under the curve for both normally and gamma distributed biomarker levels. Confidence intervals are proposed, the coverage probability of which is scrutinized by simulation study. An example using polychlorinated biphenyl levels to classify women with and without endometriosis illustrates the potential benefits of these methods. PMID- 17110641 TI - Re: "objectively measured sleep characteristics among early-middle-aged adults: the CARDIA study". PMID- 17110642 TI - Breathing-swallowing interaction in neuromuscular patients: a physiological evaluation. AB - RATIONALE: Malnutrition and aspiration are major problems in patients with neuromuscular disease. Because impaired swallowing contributes to malnutrition, means of improving swallowing are needed. OBJECTIVES: To investigate interactions between breathing and swallowing in neuromuscular disorders and to evaluate the impact of mechanical ventilation (MV) on swallowing in tracheostomized patients. METHODS: We studied 10 healthy individuals and 29 patients with neuromuscular disease and chronic respiratory failure (including 19 with tracheostomy). The tracheostomized patients who could breathe spontaneously were recorded during spontaneous breathing (SB) and with MV, in random order. MEASUREMENTS AND MAIN RESULTS: Breathing-swallowing interactions were investigated by chin electromyography and inductive respiratory plethysmography, using three water bolus sizes (5, 10, and 15 ml) in random order. In contrast to healthy individuals, neuromuscular patients showed piecemeal deglutition with several swallows over several breathing cycles for each bolus. The percentage of swallows followed by expiration was about 50% in the patients compared with nearly 100% in the control subjects. The number of swallows and total swallowing time per bolus correlated significantly to maximal inspiratory pressure. In the 10 tracheostomized patients who were recorded both in SB and MV, the number of swallows and total swallowing time per bolus were significantly reduced during MV compared with SB. CONCLUSION: Neuromuscular patients showed abnormal breathing swallowing interactions, which correlated to maximal inspiratory pressure. Moreover, MV improved the swallowing parameters in tracheostomized patients who were able to breathe spontaneously. PMID- 17110643 TI - Short-course montelukast for intermittent asthma in children: a randomized controlled trial. AB - RATIONALE: In children, intermittent asthma is the most common pattern and is responsible for the majority of exacerbations. Montelukast has a rapid onset of action and may be effective if used intermittently. OBJECTIVES: To determine whether a short course of montelukast in children with intermittent asthma would modify the severity of an asthma episode. METHODS: Children, aged 2-14 years with intermittent asthma participated in this multicenter, randomized, double-blind, placebo-controlled clinical trial over a 12-month period. Treatment with montelukast or placebo was initiated by parents at the onset of each upper respiratory tract infection or asthma symptoms and continued for a minimum of 7 days or until symptoms had resolved for 48 hours. MEASUREMENTS AND MAIN RESULTS: A total of 220 children were randomized, 107 to montelukast and 113 to placebo. There were 681 treated episodes (345 montelukast, 336 placebo) provided by 202 patients. The montelukast group had 163 unscheduled health care resource utilizations for asthma compared with 228 in the placebo group (odds ratio, 0.65; 95% confidence interval, 0.47-0.89). There was a nonsignificant reduction in specialist attendances and hospitalizations, duration of episode, and beta agonist and prednisolone use. Symptoms were reduced by 14% and nights awakened by 8.6% (p = 0.043), and days off from school or childcare by 37% and parent time off from work by 33% (p < 0.0001 for both). CONCLUSIONS: A short course of montelukast, introduced at the first signs of an asthma episode, results in a modest reduction in acute health care resource utilization, symptoms, time off from school, and parental time off from work in children with intermittent asthma. PMID- 17110644 TI - Impaired neutrophil chemotaxis in chronic obstructive pulmonary disease. AB - RATIONALE: Neutrophilic airway inflammation is considered to be a major factor in the pathogenesis of chronic obstructive pulmonary disease (COPD), with sputum and bronchoalveolar lavage neutrophil counts broadly correlating with disease severity. The mechanisms responsible for neutrophil accumulation are poorly understood, but they could involve increased influx and/or survival of these cells. OBJECTIVES: To investigate whether neutrophil chemotactic responsiveness and/or chemotactic activity in airway secretions are increased in subjects with COPD. METHODS: Chemotaxis experiments were performed using induced sputum supernatants from subjects with and without COPD as a source of chemotactic activity, and neutrophils from healthy donors as responder cells. In addition, chemotactic responses to N-formyl-Met-Leu-Phe (fMLP) and interleukin-8 (IL 8/CXCL8) were studied using neutrophils from healthy subjects and subjects with COPD. MEASUREMENTS AND MAIN RESULTS: As reported in the literature, sputum neutrophil counts were significantly increased in subjects with COPD compared with healthy subjects. However, this was associated with reduced chemotactic activity in sputum in COPD, as judged by reduced chemotaxis to the fluid phase of sputum from subjects with COPD compared with healthy subjects. Furthermore, whereas neutrophils from subjects with stage I COPD had normal responses to fMLP and IL-8, subjects with more severe stage II-IV COPD showed reduced levels of spontaneous migration and chemotaxis to fMLP and IL-8. CONCLUSIONS: Neither increased chemotactic activity in the airways nor increased chemotactic responsiveness of neutrophils explains the increased number of these cells in subjects with stable COPD. The implications of the observed reduction in neutrophil chemotactic activity remain to be established. PMID- 17110645 TI - Early-life psychological stress exacerbates adult mouse asthma via the hypothalamus-pituitary-adrenal axis. AB - RATIONALE: Despite accumulating evidence that psychological stress has a short lasting detrimental effect on asthma, little is known about the way stress in childhood predisposes to adult asthma. OBJECTIVES: Using a communication box, we investigated the long-lasting effect of early psychological and physical stress on adult asthma in mice. METHODS: Male BALB/c mice were exposed to either psychological stress or physical stress three times (every other day) during their fourth week of life. The mice were sensitized to ovalbumin at 8 and 10 weeks, and an ovalbumin airway challenge was conducted at the age of 11 weeks. RESULTS: Twenty-four hours after ovalbumin challenge, both psychological and physical stress-exposed mice exhibited a significant acceleration in the number of total mononuclear cells and eosinophils and airway hyperresponsiveness compared with control mice. No differences in serum anti-OVA-specific immunoglobulin E levels were found between stress-exposed and control animals after antigen sensitization. In the psychological stress group, but not in the physical stress group, an elevation of the serum corticosterone levels during ovalbumin challenge was significantly attenuated in comparison with the control group. Moreover, pretreatment with RU-486, a glucocorticoid receptor antagonist, before ovalbumin challenge completely inhibited a psychological stress-induced exacerbation of asthma. However, pretreatment with GR-82334, a neurokinin-1 receptor antagonist, failed to affect physical stress-induced augmentation of airway inflammation. CONCLUSION: Early psychological and physical stresses aggravated adult asthma via hyporesponsiveness of the hypothalamic-pituitary adrenal axis during antigen challenge and via a pathway(s) distinct from the hypothalamic-pituitary-adrenal axis or neurokinin-1 receptors. PMID- 17110646 TI - Burnout syndrome in critical care nursing staff. AB - RATIONALE: Burnout syndrome (BOS) associated with stress has been documented in health care professionals in many specialties. The intensive care unit (ICU) is a highly stressful environment. Little is known about BOS in critical care nursing staff. OBJECTIVES: To identify determinants of BOS in critical care nurses. METHODS: We conducted a questionnaire survey in France. Among 278 ICUs contacted for the study, 165 (59.4%) included 2,525 nursing staff members, of whom 2,392 returned questionnaires with complete Maslach Burnout Inventory data. MEASUREMENTS AND MAIN RESULTS: Of the 2,392 respondents (82% female), 80% were nurses, 15% nursing assistants, and 5% head nurses. Severe BOS-related symptoms were identified in 790 (33%) respondents. By multivariate analysis, four domains were associated with severe BOS: (1) personal characteristics, such as age (odds ratio [OR], 0.97/yr; confidence interval [CI], 0.96-0.99; p=0.0008); (2) organizational factors, such as ability to choose days off (OR, 0.69; CI, 0.52 0.91; p=0.009) or participation in an ICU research group (OR, 0.74; CI, 0.56 0.97; p=0.03); (3) quality of working relations (1-10 scale), such as conflicts with patients (OR, 1.96; CI, 1.16-1.30; p=0.01), relationship with head nurse (OR, 0.92/point; CI, 0.86-0.98; p=0.02) or physicians (OR, 0.81; CI, 0.74-0.87; p=0.0001); and (4) end-of-life related factors, such as caring for a dying patient (OR, 1.39; CI, 1.04-1.85; p=0.02), and number of decisions to forego life sustaining treatments in the last week (OR, 1.14; CI, 1.01-1.29; p=0.04). CONCLUSION: One-third of ICU nursing staff had severe BOS. Areas for improvement identified in our study include conflict prevention, participation in ICU research groups, and better management of end-of-life care. Interventional studies are needed to investigate these potentially preventive strategies. PMID- 17110647 TI - Pregnancy and chronic progressive pulmonary disease. AB - Progressive pulmonary disease may preclude the option of pregnancy for a number of women in their child-bearing years due to the severity of the disease. For a subset of women with chronic lung disease including cystic fibrosis, pregnancy is possible, but can have a devastating effect both on the prospective mother and fetus. The potential hazards of pregnancy in cystic fibrosis or other progressive pulmonary diseases may trigger a moral conflict between physician and patient. The female patient may argue that her autonomy cannot be circumscribed and that the physician is obliged to assist her reproductive efforts. The physician can counter that his/her participation in potentially harmful interventions is not consistent with professional norms requiring adherence to the principles of beneficence and nonmaleficence. Whenever possible, the ethical conflict between physician and patient should be resolved before initiation of pregnancy. We propose that this best be done through structured negotiations between physician and patient with the goal of constructing an ethical framework for reducing the moral tension between the two. Steps in the negotiating process include defining the therapeutic alliance, information exchange, dialog, and deliberation. As part of the information exchange, it is important to discuss alternatives to pregnancy such as adoption and surrogacy, especially when there are strong contraindications to pregnancy. If negotiations reach a satisfactory conclusion for both sides, there should be a well-delineated consensual agreement to commence the pregnancy with the full support of the medical team. PMID- 17110648 TI - The confounding effects of thoracic gas compression on measurement of acute bronchodilator response. AB - RATIONALE: Improvement in FEV(1) is a main endpoint in clinical trials assessing the efficacy of bronchodilators. However, the effect of bronchodilators on maximal expiratory flow may be confounded by thoracic gas compression (TGC). OBJECTIVE: To determine whether TGC confounds effect of albuterol on FEV(1). METHODS: We evaluated the response to albuterol inhalation in 10 healthy subjects, 9 subjects with asthma, and 15 subjects with chronic obstructive pulmonary disease (COPD) with mean (SD) age in years of 38 (SD, 11), 45 (SD, 11), and 64 (SD, 8), respectively. Lung mechanics were measured at baseline and 20 minutes after inhalation of 180 micro g of albuterol. We then applied a novel method to calculate FEV(1) corrected for the effect of TGC (NFEV(1)). RESULTS: Prior to albuterol administration, NFEV(1) was significantly higher than FEV(1). However, post-albuterol inhalation, FEV(1) increased more than NFEV(1) because of reduced TGC. In multiple regression analysis, the changes in TGC, inspiratory lung resistance, and ratio of residual volume to total lung capacity postalbuterol predicted more than 75% of FEV(1) improvement in patients with COPD. CONCLUSION: Improvements in FEV(1) after albuterol in patients with COPD are due to reduction of lung resistance, hyperinflation, and TGC. The latter is negligible during tidal breathing. Thus, although reduction of lung resistance and hyperinflation may result in improved dyspnea with a bronchodilator, the contribution of TGC reduction to improvement of FEV(1) may not exert any meaningful clinical effect during tidal breathing. This fact has to be taken into consideration when assessing the efficacy of new bronchodilators. PMID- 17110649 TI - Comparison of biomarkers in exhaled breath condensate and bronchoalveolar lavage. AB - RATIONALE: Exhaled breath condensate (EBC) is increasingly studied as a noninvasive research method of sampling the lungs, measuring several biomarkers. The exact site of origin of substances measured in EBC is unknown, as is the clinical applicability of the technique. Special techniques might be needed to measure EBC biomarkers. OBJECTIVES: To assess biomarker concentrations in clinical disease and investigate the site of origin of EBC, we compared EBC and bronchoalveolar lavage (BAL) biomarkers in 49 patients undergoing bronchoscopy for clinical indications. MEASUREMENTS: We measured exhaled nitric oxide, 8 isoprostane, hydrogen peroxide, total nitrogen oxides, pH, total protein, and phospholipid (n = 33) and keratin (n = 15) to assess alveolar and mucinous compartments, respectively. EBC was collected over 10 min using a refrigerated condenser according to European Respiratory Society/American Thoracic Society recommendations, and BAL performed immediately thereafter. RESULTS: 8 Isoprostane, nitrogen oxides, and pH were significantly higher in EBC than in BAL (3.845 vs. 0.027 ng/ml, 28.4 vs. 3.8 microM, and 7.35 vs. 6.4, respectively; p < 0.001). Hydrogen peroxide showed no difference between EBC and BAL (17.5 vs. 20.6 microM, p = not significant), whereas protein was significantly higher in BAL (33.8 vs. 183.2 microg/ml, p < 0.001). Total phospholipid was also higher in EBC, but keratin showed no difference. No significant correlation was found between EBC and BAL for any of the biomarkers evaluated either before or after correction for dilution. CONCLUSIONS: In clinical disease, markers of inflammation and oxidative stress are easily measurable in EBC using standard laboratory techniques and EBC is readily obtained. However, EBC and BAL markers do not correlate. PMID- 17110650 TI - Reducing the cost of critical care: new challenges, new solutions. PMID- 17110651 TI - Where there's fire, there's smoke. PMID- 17110652 TI - Decision rules and pneumonia: What are we "predicting," and for whom? PMID- 17110653 TI - Animal models of asthma: Pro: Allergic avoidance of animal (model[s]) is not an option. PMID- 17110654 TI - The mouse trap: It still yields few answers in asthma. PMID- 17110657 TI - Noninvasive ventilation versus nonrebreather bag-valve mask to achieve preoxygenation before intubation of hypoxic patients. PMID- 17110658 TI - 2006: a stroke odyssey. PMID- 17110659 TI - An observation of interest relative to the practice of spectroscopic measurements in multiple sclerosis. PMID- 17110660 TI - Correction of HEAL Registry Data Report. PMID- 17110661 TI - Resectability issues with head and neck cancer. AB - Head and neck surgeons often rely on imaging to determine if a neoplasm is resectable. Many of the critical issues are outlined in the American Joint Committee on Cancer Staging Manual, wherein T4a and T4b head and neck cancers are defined as resectable and unresectable, respectively. Even within the T4a advanced resectable classification, there are critical determinants that define whether the surgical option is such that major morbidity and mortality could be expected. This review article examines the imaging literature to determine the accuracy and diagnostic criteria of different modalities for evaluating these critical T4a and T4b factors, which include the following: 1) arterial encasement, 2) prevertebral fascia involvement, 3) mediastinal infiltration, 4) tracheal and esophageal extension, 5) laryngeal cartilage penetration, 6) pre epiglottic fat involvement, 7) dural spread, 8) bone (mandible/maxilla and skull base) infiltration, 9) perineural spread, 10) orbital involvement, and 11) brachial plexus invasion. For the most part, the studies find MR imaging with higher sensitivity but lower specificity than CT. An ever-increasing role for PET/CT is suggested. Imaging is of great value in the determination of resectability issues listed previously for head and neck cancers, with the possible exception of prevertebral fascia involvement. PMID- 17110662 TI - Neuropathology for the neuroradiologist: palisades and pseudopalisades. AB - Histologic patterns of cellular architecture often suggest a tissue diagnosis. One distinctive histologic pattern seen within some tumors of the nervous system is the palisade. The purpose of this report is to review the significance of palisades and pseudopalisades in the context of such tumors as schwannomas and glioblastomas. PMID- 17110663 TI - Outcome of acute vertebrobasilar occlusions treated with intra-arterial fibrinolysis in 180 patients. AB - BACKGROUND AND PURPOSE: To evaluate predictors of recanalization and a favorable neurologic outcome in patients with acute vertebrobasilar occlusion (VBO) treated with local intra-arterial fibrinolysis (LIF). METHODS: The multicentric data of 180 patients with acute VBO treated with LIF were retrospectively evaluated. The modified Rankin scale (mRS) was used to evaluate the neurologic status before LIF and at the time of discharge. Patient's sex, age, etiology of VBO, recanalization, symptom duration before LIF, and pretreatment mRS were correlated with posttreatment mRS. Multiple logistic regression analysis was used to identify independent variables for recanalization and neurologic outcome. RESULTS: The overall mortality was 43%. Complete recanalization was achieved in 99 (55%) patients and a partial recanalization in 35 (19%) patients, respectively. Recanalization was significantly associated with a favorable outcome (P < .001). The success of recanalization was negatively correlated with the volume of the thrombus (P < .001). No correlation was found between site and etiology of VBO and recanalization. Neurologic outcome correlated strongly with the pretreatment mRS (P < .001) and also with age (P < .02). Coma lasting less than 4.5 hours led to a positive trend toward a better outcome after univariate testing (P < .001). CONCLUSIONS: Success of recanalization and neurologic status before treatment predict neurologic outcome in patients with VBO. Thrombus volume has an adverse effect on the recanalization success. PMID- 17110664 TI - Endovascular mechanical clot retrieval in a broad ischemic stroke cohort. AB - BACKGROUND AND PURPOSE: Our aim was to describe an expanded experience with endovascular mechanical embolectomy in a broad group of patients, including those not meeting entry criteria for the MERCI multicenter trials. METHODS: We performed an analysis of all patients with ischemic stroke treated with the Merci Clot Retrieval Device at a single academic center outside of the Mechanical Embolus Removal in Cerebral Ischemia (MERCI) trials. RESULTS: Twenty-four patients were treated with the device. Nine were MERCI trial ineligible: 4 received intravenous (IV) tissue plasminogen activator (tPA), 1 received IV tPA and was younger than 18 years of age, and 4 had time-to-treatment of longer than 8 hours. Mean age was 64 years (range, 14-89 years; 42% women). Median National Institutes of Health Stroke Scale (NIHSS) score was 21 (range, 11-30). Median symptoms-to-procedure-start time was 303 minutes (range, 85-2385 minutes). Recanalization (Thrombolysis in Myocardial Infarction, 2-3) was achieved in 15/24 (63%). In device-only patients, recanalization occurred in 10/16. In patients who failed IV tPA undergoing rescue embolectomy, recanalization was achieved in 4/5. Three patients unresponsive to device therapy received rescue intra-arterial tPA/abciximab; recanalization was achieved in 2/3. Recanalization was achieved in 3/4 patients in whom treatment was started longer than 8 hours after symptom onset. Asymptomatic hemorrhage occurred in 38%; symptomatic hemorrhage, in 8%. Three device fractures occurred; none worsened clinical outcome. In-hospital mortality was 17%; 90-day mortality, 29%. Good 90-day functional outcome (modified Rankin Scale, 30 days), we saw the resolution of signal intensity abnormalities and the appearance of atrophic change (4 of 4, 100%) of the affected regions. CONCLUSION: HHV-6-associated encephalopathy in adults tends to affect the mesial temporal lobe. MR imaging is useful for detecting HHV-6 encephalopathy and distinguishing it from the other diseases of the central nervous system in immunocompromised patients. PMID- 17110692 TI - MR features of intraocular ectopic lacrimal tissue. AB - A 2-year-old girl who had a 3-day history of swelling in her right eye presented with a case of intraocular ectopic lacrimal tissue. MR imaging findings and possible causes of the ectopic lacrimal tissue in the globe, including embryology, are reviewed. The differential diagnoses of other intraocular masses and the imaging features that can be of help in making a diagnosis are also discussed. PMID- 17110693 TI - Metachronous, multicentric giant cell tumor of the sphenoid bone with histologic, CT, MR imaging, and positron-emission tomography/CT correlation. AB - Giant cell tumor (GCT) of the sphenoid bone is a relatively rare entity and metachronous multicentric GCT of the sphenoid is even rarer; we are aware of only 3 previous cases in the literature. We describe here a tumor of the sphenoid bone that was identified 15 years after multiple resections of a GCT of the left inferior pubic ramus. Correlation is made between the histopathologic findings, MR imaging of the brain, CT of the head, and fusion positron-emission tomography (PET)/CT scan performed with fluorine-18 fluoro-2-deoxy-D-glucose (18F-FDG). This report is the first to describe the appearance of a GCT of the sphenoid bone on a fusion PET/CT examination. High metabolic activity in the base of the skull adjacent to the middle cranial fossa was demonstrated in a fashion similar to that of the known pelvic lesion. This case also demonstrates that the increased metabolic activity seen in a GCT of the sphenoid bone may be partially obscured by the adjacent physiologic high metabolic activity of the brain. PMID- 17110694 TI - Tongue abscess mimicking neoplasia. AB - Tongue abscess is a very rare condition about which only sparse imaging findings have been reported. In 2 patients, a submucosal malignant tumor was suspected because of the presence of a painful hard tongue mass with intact mucosal surface. Differential diagnosis of the tongue abscess or malignancy was difficult on MR imaging or positron emission tomography-CT. In careful retrospective history taking, the symptoms had dated from an episode of eating fish several months before presentation. Plain radiographs and noncontrast CT images of the pharynx revealed a foreign body within the lesion. Thus, we present the imaging findings of 2 cases of malignancy mimicking tongue abscess as a result of impaction of a fish bone. PMID- 17110695 TI - Imaging and clinical characteristics of temporal bone meningioma. AB - BACKGROUND AND PURPOSE: Imaging characteristics of temporal bone meningioma have not been previously reported in the literature. CT and MR imaging findings in 13 cases of temporal bone meningioma are reviewed to define specific imaging features. METHODS: A retrospective review of our institutional case archive revealed 13 cases of histologically confirmed temporal bone meningioma. CT and MR imaging studies were reviewed to characterize mass location, vector of spread, bone changes, enhancement characteristics, and intracranial patterns of involvement. Clinical presenting signs and symptoms were correlated with imaging findings. RESULTS: Thirteen temporal bone meningiomas were reviewed in 8 women and 5 men, aged 18-65 years. Meningiomas were stratified into 3 groups on the basis of location and tumor vector of spread. There were 6 tegmen tympani, 5 jugular foramen (JF), and 2 internal auditory canal (IAC) meningiomas. Tegmen tympani and JF meningiomas were characterized by spread to the middle ear cavity. IAC meningiomas, by contrast, spread to the cochlea and vestibule. Hearing loss was the most common clinical presenting feature in all cases of temporal bone meningioma (10/13). The presence of tumor adjacent to the ossicles strongly correlated with conductive hearing loss (7/9). CONCLUSION: Meningioma involving the temporal bone is rare. Three subgroups of meningioma exist in this location: tegmen tympani, JF, and IAC meningioma. Tegmen tympani and JF meningiomas spread to the middle ear cavity. IAC meningiomas spread to intralabyrinthine structures. Conductive hearing loss is commonly seen in these patients and can be surgically correctable. PMID- 17110696 TI - MR spectroscopy in sinus mucocele: N-acetyl mimics of brain N-acetylaspartate. AB - We describe MR spectroscopy in 2 patients with frontal sinus mucoceles that showed a dominant metabolite peak at 2.0-ppm chemical shift, simulating N acetylaspartate (NAA) of normal neuronal tissue. In vitro analysis of postsurgical mucocele samples confirmed that the signal at 2.0 ppm was arising from the methyl moiety of an N-acetyl compound. This is probably caused by N acetylgalactosamine or N-acetylglucosamine, which are glycoproteins found in normal respiratory mucus produced by the paranasal sinus epithelium. PMID- 17110697 TI - Unilateral submandibular gland aplasia associated with ipsilateral sublingual gland hypertrophy. AB - Congenital absence of the major salivary glands is an infrequent disorder. Clinically, patients may be asymptomatic or may present with dryness of the mouth, difficulty in chewing and swallowing, and dental caries. The absence of the submandibular gland may be associated with hypertrophy of the contralateral submandibular gland. We report a case of ipsilateral sublingual hypertrophy associated with unilateral submandibular aplasia. PMID- 17110698 TI - Inflammatory myofibroblastic tumor of the orbit with associated enhancement of the meninges and multiple cranial nerves. AB - Inflammatory myofibroblastic tumor (IMT), Tolosa-Hunt syndrome (THS), and idiopathic hypertrophic pachymeningitis (IHP) seem to be part of a spectrum of disorders that have diverse locations but similar histologic and imaging findings. We report a case of a 50-year-old man presenting with multiple progressive cranial nerves palsies with leptomeningeal cranial nerve enhancement on MRI (II, V1-V3, and X), orbital and infraorbital masses, prominence within the left cavernous sinus, and diffuse dural enhancement. Biopsies of the orbital lesion and infraorbital nerve revealed IMT. The patient's lesions, symptoms, and dural enhancement quickly improved with steroid administration and nearly resolved over multiple subsequent scans over the next few months. This case illustrates a rare case of pseudotumor mimicking a more aggressive appearance that would usually portend a case of malignancy. There is a potential association of IMT, THS, and IHP, which may have existed in a concomitant fashion in this patient. The case also describes the unique finding of enhancement of the cisternal segments of multiple cranial nerves (simulating leptomeningeal malignant involvement), which may be related to inflammatory perineural edema or ischemic neuropathy. PMID- 17110699 TI - Optimization of Z-axis automatic exposure control for multidetector row CT evaluation of neck and comparison with fixed tube current technique for image quality and radiation dose. AB - BACKGROUND AND PURPOSE: Z-axis automatic exposure control (AEC) technique automatically adjusts tube current based on size of body region scanned. The purpose of the current study was to compare diagnostic acceptability, noise, and radiation exposure of multidetector row CT (MDCT) of neck performed with z-axis AEC and with fixed current. MATERIALS AND METHODS: Two study groups of 26 patients each underwent MDCT of neck using z-axis AEC with 8 noise index (NI), 150-440 mA, and 10 NI, 75-440 mA, respectively. A control group consisting of another 26 patients underwent MDCT of neck with fixed-current technique (300 mA). Objective noise and mean tube current-time products (mA . s) were recorded. Two radiologists evaluated images for diagnostic acceptability and subjective noise on a 5-point scale. RESULTS: All CT examinations of study and control groups were diagnostically acceptable, though objective noise was significantly more with z axis AEC (shoulder: NI 8, 20.6 +/- 6.2 HU; NI 10, 22.2 +/- 4.6 HU) than with fixed current (16.2 +/- 6 HU) (P = .01). There was no significant difference between AEC and fixed current in diagnostic acceptability and subjective noise (P = .22-.42). AEC resulted in significant radiation dose reduction (NI 8, 186.3 +/- 20.5 mA . s; NI 10, 158.1 +/- 21.2 mA x s), compared with fixed current (235 +/- 21.8 mA x s). CONCLUSION: Z-axis AEC resulted in similar subjective noise and diagnostic acceptability, with radiation dose reduction of 21% for NI of 8 and 33% for NI of 10, respectively, for MDCT evaluation of neck, compared with those of fixed current technique. PMID- 17110700 TI - Resuscitation using albumin in critically ill patients: research in patients at high risk of complications is now needed. PMID- 17110701 TI - Glomerular filtration rate: screening cannot be recommended on the basis of current knowledge. PMID- 17110702 TI - Tackling childhood obesity: requires a shift in social norms, not just an exercise programme. PMID- 17110703 TI - Ethics group rules on treating premature babies. PMID- 17110704 TI - Dutch insurers pay midwives not to refer breech presentations to hospital. PMID- 17110705 TI - US health industry to set safety guides for hospitals. PMID- 17110706 TI - GMC strikes off expert in drug addiction. PMID- 17110707 TI - Royal College of Surgeons give cautious approval for face transplants. PMID- 17110709 TI - Indian health activists criticise voluntary price cuts by drug industry. PMID- 17110708 TI - Doha Declaration has failed to deliver cheap drugs to developing countries, Oxfam says. PMID- 17110710 TI - European health commissioner praises food firms for efforts to cut obesity. PMID- 17110713 TI - Woman dies after doctors fail to intervene because of new abortion law in Nicaragua. PMID- 17110714 TI - High Court judge over-rules GMC decision not to proceed against GP. PMID- 17110715 TI - Charity demands better alcohol treatment services as death rates double. PMID- 17110717 TI - Law to resolve small negligence claims gets royal assent. PMID- 17110718 TI - A new direction for health? PMID- 17110720 TI - Greater collaboration needed to reduce global burden of disease. PMID- 17110719 TI - South Asia struggles to meet targets on health and hunger. PMID- 17110721 TI - Australian Senate votes to allow therapeutic cloning. PMID- 17110722 TI - Who's using whom? PMID- 17110723 TI - Chronic constipation in children. PMID- 17110724 TI - Methadone tolerance testing in drug misusers. PMID- 17110725 TI - ABC of obesity: obesity and vascular disease. PMID- 17110726 TI - Refusing to provide a prenatal test: can it ever be ethical? PMID- 17110727 TI - Analogies between reading of medical and religious texts. PMID- 17110728 TI - Reactions to the chief medical officer's report: overegging the pudding. PMID- 17110729 TI - Reactions to the chief medical officer's report: less attractive than death or taxes. PMID- 17110730 TI - Reactions to the chief medical officer's report: the GMC has changed fundamentally. PMID- 17110731 TI - How to measure renal function in clinical practice: stage 3 chronic kidney disease is not a consequence of normal ageing. PMID- 17110732 TI - How to measure renal function in clinical practice: eating cooked meat alters serum creatinine concentration and eGFR significantly. PMID- 17110733 TI - Reactions to the chief medical officer's report: where are patients' voices? PMID- 17110734 TI - Scotland v England deal on prescribed drugs: Scottish Medicines Consortium responds. PMID- 17110735 TI - Delayed cord clamping may also be beneficial in rich settings. PMID- 17110736 TI - Roger Sperry: pioneer of neuronal specificity. AB - This essay looks at the historical significance of two APS classic papers that are freely available online: Sperry RW. Optic nerve regeneration with return of vision in anurans. J Neurophysiol 7: 57-69, 1944 (http://jn.physiology.org/cgi/reprint/7/1/57). Sperry RW. Restoration of vision after crossing of optic nerves and after contralateral transplantation of eye. J Neurophysiol 8: 15-28, 1945 (http://jn.physiology.org/cgi/reprint/8/1/15). PMID- 17110737 TI - Language and the aging brain: patterns of neural compensation revealed by functional brain imaging. AB - Human aging brings with it declines in sensory function, both in vision and in hearing, as well as a general slowing in a variety of perceptual and cognitive operations. Yet in spite of these declines, language comprehension typically remains well preserved in normal aging. We review data from functional magnetic resonance imaging (fMRI) to describe a two-component model of sentence comprehension: a core sentence-processing area located in the perisylvian region of the left cerebral hemisphere and an associated network of brain regions that support the working memory and other resources needed for comprehension of long or syntactically complex sentences. We use this two-component model to describe the nature of compensatory recruitment of novel brain regions observed when healthy older adults show the same success at comprehending sentences as their younger adult counterparts. We suggest that this plasticity in neural recruitment contributes to the stability of language comprehension in the aging brain. PMID- 17110738 TI - Time-dependent, layer-specific modulation of sensory responses mediated by neocortical layer 1. AB - An essential component of feedback and top-down information in the cortical column arrives at layer 1 (L1) where it contacts distal dendrites of pyramidal neurons. Although much is known about the anatomical organization of L1 fibers, their contribution to sensory information processing remains to be determined. We assessed the physiological significance of L1 inputs by performing extracellular recordings in vivo from neurons in the primary somatosensory cortex of rodents. We found that blocking activity in L1 increases whisker-evoked response magnitude and variance, suggesting that L1 exerts an inhibitory influence on whisker responses. However, when pairing L1 stimulation with whisker deflection, the interval between the stimuli determined the outcome of the interaction, with facilitation of sensory responses dominating the short intervals (10 ms). These temporal interactions resulted in a time-dependent regulation of direction tuning of cortical neurons. The synaptic mechanisms underlying L1 inputs' influences were examined using whole cell recordings in vitro while pairing L1 and white-matter stimulations. We found time-dependent, layer-specific differences in synaptic summation of the two inputs, with supralinearity at shorter intervals and sublinearity at longer intervals that resulted mainly from shunting inhibition. Taken together, our results demonstrate that L1 inputs impose a time- and layer-specific regulation on sensory-evoked responses. This in turn may lead to a dynamic transmission of sensory information in the somatosensory cortex. PMID- 17110739 TI - Identification of basolateral amygdala projection cells and interneurons using extracellular recordings. AB - This study tested whether firing rate and spike shape could be used to distinguish projection cells from interneurons in extracellular recordings of basolateral amygdala (BLA) neurons. To this end, we recorded BLA neurons in isoflurane-anesthetized animals with tungsten microelectrodes. Projection cells were identified by antidromic activation from cortical projection sites of the BLA. Although most projection cells fired spontaneously at low rates (<1 Hz), an important subset fired at higher rates (up to 6.8 Hz). In fact, the distribution of firing rates in projection cells and unidentified BLA neurons overlapped extensively, even though the latter cell group presumably contains a higher proportion of interneurons. The only difference between the two distributions was a small subset (5.1%) of unidentified neurons with unusually high firing rates (9 16 Hz). Similarly, distributions of spike durations in both cell groups were indistinguishable, although most of the fast-firing neurons had spike durations at the low end of the distribution. However, we observed that spike durations depended on the exact position of the electrode with respect to the recorded cell, varying by as much as 0.7 ms. Thus neither firing rate nor spike waveform allowed for unequivocal separation of projection cells from interneurons. Nevertheless, we propose the use of two firing rate cutoffs to obtain relatively pure samples of projection cells and interneurons: < or =1 Hz for projection cells and > or =7 Hz for fast-spiking interneurons. Supplemented with spike duration cutoffs of > or =0.7 ms for projection cells and < or =0.5 ms for interneurons, this approach should keep instances of misclassifications to a minimum. PMID- 17110740 TI - Presynaptic plasticity in an immature neocortical network requires NMDA receptor activation and BDNF release. AB - Activity-dependent developmental maturation of the neocortical network is thought to involve the stabilization and potentiation of immature synapses. In particular, N-methyl-d-aspartate (NMDA) receptor-dependent long-term plasticity that is expressed presynaptically appears to be crucial for the selection of functionally adequate synapses. However, presynaptic expression of long-term plasticity in neocortical neurons has mainly been studied indirectly by electrophysiological techniques. Here we analyzed presynaptic plasticity directly by repeated imaging of actively cycling presynaptic vesicles with the styryl dye FM4-64 in cultured neocortical neurons at 34 degrees C. To monitor long-term changes, stimulation-induced saturating FM4-64 staining and subsequent destaining was performed twice with an interval of 1.5 h between stainings and with the first staining serving as a plasticity stimulus. In the vast majority of presynaptic release sites, we found an increase in the mean fluorescence intensity after the second staining indicating an enhanced number of cycling synaptic vesicles. Most intriguingly, we additionally observed the appearance of new active release sites. As demonstrated by the addition of the NMDA receptor antagonist d-2-amino-5-phosphonopentanoic acid (d-AP5), both plasticity phenomena were strictly dependent on NMDA receptor activation. This suggests that a subpopulation of release sites was functionally silent during the first round of staining. Moreover, we studied a potential role of brain-derived neurotrophic factor (BDNF) in this type of presynaptic plasticity by imaging BDNF-deficient neocortical neurons. The increase in fluorescence intensity was strongly inhibited in BDNF-knockout neurons and was absent in wild-type neurons in the presence of BDNF scavenging trkB receptor bodies. These results indicate that BDNF might play an important role as a plasticity-related messenger molecule in neocortical neurons. PMID- 17110741 TI - Understanding the physiopathology of paraneoplastic and genetic cerebellar ataxia. PMID- 17110742 TI - Ten steps to identify atypical parkinsonism. PMID- 17110743 TI - Prevention of delayed cerebral ischaemia after subarachnoid haemorrhage. PMID- 17110744 TI - Genetic association studies of complex neurological diseases. AB - Genetic association studies offer a powerful approach to identify the multiple variants of small effect that modulate susceptibility to common, complex disease. They, however, have a poor reputation, mainly because of the consistent lack of replication of all but a few. Thousands of genetic studies have been carried out on multifactorial diseases in the past 30 years, yielding only about 50 variants that can be considered to be true positives. Although the positive studies show proof of principle, the multitude of negative studies indicate fundamental problems in the design and execution of association studies. Here, we discuss some of the more pertinent study design and data analysis issues which can affect the outcome of genetic association studies. PMID- 17110745 TI - Referral guidelines for suspected central nervous system or brain tumours. PMID- 17110746 TI - Von Frey's pain spots. PMID- 17110747 TI - Neurological picture. Extensive brain calcification in idiopathic hypoparathyroidism. PMID- 17110748 TI - Neurological picture. Susac's syndrome: effective combination of immunosuppression and antiplatelet treatment. PMID- 17110749 TI - Manic behaviour induced by deep-brain stimulation in Parkinson's disease: evidence of substantia nigra implication? AB - We report the case of a patient who had benefited from bilateral subthalamic nucleus deep brain stimulation for Parkinson's disease and who presented acute and reproducible manic behaviour when stimulated mainly in the substantia nigra. A positron emission tomography scan showed an activation of the right dorsolateral prefrontal and inferior temporal cortex, the left anterior cingulate cortex and a deactivation of the left insula. This suggests that changes in cortical activation related to mania are subcortically driven, involving notably the substantia nigra. PMID- 17110750 TI - Clinical heterogeneity of recessive ataxia in the Mexican population. AB - Approximately 75% of Indo-European patients with recessive ataxia are homozygous for frataxin gene (FXN) mutations and have either typical or atypical Friedreich ataxia (FRDA). Our previous analysis of 134 Mexican Mestizo recessive ataxia patients showed that FRDA is relatively uncommon in the Mexican population (10.4%). This article reports the evaluation of the phenotypes of these patients. Over half of the patients with clinical diagnostic criteria for FRDA did not carry FXN mutations, constituting a "FRDA-like" phenotypic subgroup. Analysis of non-FRDA patients revealed a subgroup with early onset recessive cerebellar ataxia and cognitive deficit. These two phenotypic subgroups accounted for approximately 60% of all patients, indicating that the cause for recessive ataxia in the Mexican population is distinct from other populations and remains largely unknown. PMID- 17110751 TI - Abnormal pontine activation in pathological laughing as shown by functional magnetic resonance imaging. AB - To explore the aetiology of pathological laughing, a 65-year-old woman with pathological laughing was examined by 3-T functional magnetic resonance imaging (fMRI) before and after treatment with drugs. Here, we report that the patient consistently showed exaggerated pontine activation during the performance of three tasks before treatment, whereas abnormal pontine activation was no longer found after successful treatment with the selective serotonin reuptake inhibitor, paroxetine. Our findings in this first fMRI study of pathological laughing suggest that serotonergic replacement decreases the aberrant activity in a circuit that involves the pons. PMID- 17110752 TI - Why a positive genetic test for myotonic dystrophy type I does not always imply the right diagnosis. PMID- 17110753 TI - Selective brain activity in response to one's own name in the persistent vegetative state. PMID- 17110754 TI - Intracranial hypertension causing polyradiculopathy and late or absent F-waves. PMID- 17110757 TI - Dose area product evaluations with Gafchromic XR-R films and a flat-bed scanner. AB - Gafchromic XR-R films are a useful tool to evaluate entrance skin dose in interventional radiology. Another dosimetric quantity of interest in diagnostic and interventional radiology is the dose area product (DAP). In this study, a method to evaluate DAP using Gafchromic XR-R films and a flat-bed scanner was developed and tested. Film samples were exposed to an x-ray beam of 80 kVp over a dose range of 0-10 Gy. DAP measurements with films were obtained from the digitalization of a film sample positioned over the x-ray beam window during the exposure. DAP values obtained with this method were compared for 23 cardiological interventional procedures with DAP values displayed by the equipment. The overall one-sigma dose measurement uncertainty depended on the absorbed dose, with values below 6% for doses above 1 Gy. A maximum discrepancy of 16% was found, which is of the order of the differences in the DAP measurements that may occur with different calibration procedures. Based on the results presented, after an accurate calibration procedure and a thorough inspection of the relationship between the actual dose and the direct measured quantity (net optical density or net pixel value variation), Gafchromic XR-R films can be used to assess the DAP. PMID- 17110758 TI - Variations in calculated SAR with distance to the perfectly matched layer boundary for a human voxel model. AB - The purpose of this work was to investigate the dependence of whole-body averaged specific energy absorption rate (SAR), calculated using the finite-difference time-domain (FDTD) method, on the width of the free space region between a numerical phantom and the perfectly matched layer (pml) absorbing boundary. Results show that an increase in this width from 2 cells to 70 cells caused variations in the calculated whole-body averaged SAR of less than 2% for the FDTD code employing split-field pmls. Similarly, an increase in the width of the pml layer had little effect on the whole-body SAR values. PMID- 17110759 TI - Visible absorption spectra of radiation exposed SIRAD dosimeters. AB - SIRAD badge dosimeters are a new type of personal dosimeter designed to measure radiation exposure up to 200 R and give a visual qualitative measurement of exposure. This is performed using the active dosimeter window, which contains a radiochromic material amalgamated in the badge assembly. When irradiated, the badges active window turns blue, which can be matched against the given colour chart for a qualitative assessment of the exposure received. Measurements have been performed to analyse the absorption spectra of the active window, and results show that the window automatically turns a blue colour upon irradiation and produces two peaks in the absorption spectra located at 617 nm and 567 nm. When analysed with a common computer desktop scanner, the optical density response of the film to radiation exposure is non-linear but reproducible. The net OD of the film was 0.21 at 50 R exposure and 0.31 at 200 R exposure when irradiated with a 6 MV x-ray energy beam. When compared to the calibration colour strips at 6 MV x-ray energy the film's OD response matches relatively well within 3.5%. An approximate 8% reduction in measured OD to exposure was seen for 250 kVp x-rays compared to 6 MV x-rays. The film provides an adequate measurement and visually qualitative assessment of radiation exposure for levels in the range of 0 to 200 R. PMID- 17110760 TI - Relating two techniques for handling uncertainties in IMRT optimization. AB - Radiotherapy treatment planning is associated with uncertainties. Examples are uncertainties in the tumour location due to organ movement or the inter/intra observer variability in target definition. Different approaches to incorporate uncertainties into IMRT optimization have been proposed. In this note, we point out a relation between two previously published methods: the coverage probability approach and the concept of optimizing the expectation value of an objective function that depends on a set of random variables. Both concepts are generally different, but turn out to be equivalent in special cases. PMID- 17110761 TI - Optical devices used for image analysis of pigmented skin lesions: a proposal for quality assurance protocol using tissue-like phantoms. AB - Different technological tools have been developed to aid in the diagnosis of pigmented skin lesions, including cameras working with conventional RGB colour systems, epiluminescence microscopy and spectrophotometric methods using visible and near infrared wavelengths. All the different procedures should provide in an objective and reproducible fashion quantitative measurements of the colour and shape features of a given skin mole. At present, many devices have been introduced in experimental stages for clinical diagnosis, mainly used to provide to the clinicians an objective, computer-assisted second opinion. As for any diagnostic instruments, optical devices should also be subjected to a dedicated quality assurance protocol in order to evaluate the response repeatability of each device (intra-instrument agreement) and to check the accordance among the responses of different devices (inter-instrument agreement). The aim of this study was to design a quality assurance protocol for optical devices dedicated to image analysis of pigmented skin lesions and, in case, to detect cutaneous melanoma by using suitable tissue-like phantoms as standard references that enable testing of both hardware and software components. As an example, we report the results of intra-instrument and inter-instrument agreement when the protocol was applied on a series of 30 SpectroShade instruments, a novel optical device based on multi-spectral image analysis of colour and shape features of pigmented skin lesion. PMID- 17110762 TI - Coordinate transformations for BEAM/EGSnrc Monte Carlo dose calculations of non coplanar fields received from a DICOM-compliant treatment planning system. AB - The Monte Carlo (MC) method provides the most accurate to-date dose calculations in heterogeneous media and complex geometries, and this spawns increasing interest in incorporating MC calculations in the treatment planning quality assurance process. This process involves MC dose calculations for the treatment plans produced clinically. Commonly used in radiotherapy, MC codes are BEAMnrc and DOSXYZnrc, which transport particles in a coordinate system (c.s.) that has been established historically and does not correspond to the c.s. of treatment planning systems (TPSs). Relative rotations of these c.s. are not straightforward, especially for non-coplanar treatments. Transformation equations are therefore required to re-calculate a treatment plan using BEAM/DOSXYZnrc codes. This paper presents such transformations for beam angles defined in a DICOM-compliant treatment planning coordinate system. Verification of the derived transformations with two three-field plans simulated on a phantom using TPS as well as MC codes has been performed demonstrating exact geometrical agreement of the MC treatment fields' placement. PMID- 17110763 TI - Towards conformal light delivery using tailored cylindrical diffusers: attainable light dose distributions. AB - Interstitial light delivery for therapeutic applications requires the use of fibre-based light diffusers. Such diffusers are presently manufactured to emit with a flat longitudinal power profile. Recently, diffusers with tailored longitudinal emission profiles have become available opening an avenue to improve conformal light delivery. This paper explores the ability of tailored diffusers to improve light dose confinement to the target volume. A formalism to calculate the light dose from an arbitrary source distribution is presented based on the convolution with an appropriate point source function. By choosing a source distribution corresponding to a cylindrical diffuser emitting with a sinusoidal profile, the set of attainable light dose distributions is characterized via a relationship between the diffuser's spatial frequency, the radial distance and the amplitude of the isodose contour. PMID- 17110764 TI - Beta dosimetry with microMOSFETs for endovascular brachytherapy. AB - The aim of this study was to investigate if microMOSFETs are suitable for the dosimetry and quality assurance of beta sources. The microMOSFET dosimeters have been tested for their angular dependence in a 6 MeV electron beam. The dose rate dependence was measured with an iridium-192 afterloading source. By varying the source-to-surface distance (SSD) in a 12 MeV electron beam the dose rate dependence in an electron beam was also investigated. To measure a depth dose curve the dose rate at 2, 5, 8 and 12 mm distance from the beta source train axis was determined with the OPTIDOS and the microMOSFET detector. A comparison between the two detector types shows that the microMOSFET is suitable for quality assurance of beta sources for endovascular brachytherapy (EVBT). The homogeneity of the source is checked by measurements at five points (for the 60 mm source at 10, 20, 30, 40 and 50 mm) along the source train. The microMOSFET was then used to evaluate the influence of a common stent type (single layer stainless steel) on the dose distribution in water. The stent led to a dose inhomogeneity of +/ 8.5%. Additionally the percentage depth dose curves with and without a stent were compared. The depth dose curves show good agreement which means that the stent does not change the beta spectrum significantly. PMID- 17110765 TI - Design of a computer-controlled multileaf collimator for advanced electron radiotherapy. AB - A multileaf collimator for electrons (eMLC) has been designed that fulfils the technical requirements for providing advanced irradiation techniques with electrons. In the present work, the basic design parameters of leaf material, leaf height, leaf width and number of leaves as well as leaf overtravel and leaf shape were determined such that an eMLC with motorized leaves can be manufactured by a company specialized in MLC technology. For this purpose, a manually driven eMLC with variable source-to-collimator distance (SCD) was used to evaluate the chosen leaf specification and investigate the impact of the SCD on the off-axis dose distribution. In order to select the final SCD of the eMLC, a compromise had to be found between maximum field size, minimum beam penumbra and necessary distance between eMLC and isocentre to eliminate patient realignments during gantry rotation. As a result, the eMLC is placed according to the target position at 72 and 84 cm SCD, respectively. This feature will be achieved by interchangeable distance holders. At these SCDs, the corresponding maximum field sizes at 100 cm source-to-isocentre distance are 20 x 20 cm and 17 x 17 cm, respectively. Finally, the off-axis dose distribution at the maximum opening of the eMLC was improved by fine-tuning the settings of the accelerator jaws and introducing trimmer bars above the eMLC. Following this optimization, a prototype eMLC consisting of 2 x 24 computer-controlled brass leaves is manufactured by 3D Line Medical Systems. PMID- 17110766 TI - A technique to consider mismatches between fMRI and EEG/MEG sources for fMRI constrained EEG/MEG source imaging: a preliminary simulation study. AB - fMRI-constrained EEG/MEG source imaging can be a powerful tool in studying human brain functions with enhanced spatial and temporal resolutions. Recent studies on the combination of fMRI and EEG/MEG have suggested that fMRI prior information could be readily implemented by simply imposing different weighting factors to cortical sources overlapping with the fMRI activations. It has been also reported, however, that such a hard constraint may cause severe distortions or elimination of meaningful EEG/MEG sources when there are distinct mismatches between the fMRI activations and the EEG/MEG sources. If one wants to obtain the actual EEG/MEG source locations and uses the fMRI prior information as just an auxiliary tool to enhance focality of the distributed EEG/MEG sources, it is reasonable to weaken the strength of fMRI constraint when severe mismatches between fMRI and EEG/MEG sources are observed. The present study suggests an efficient technique to automatically adjust the strength of fMRI constraint according to the mismatch level. The use of the proposed technique rarely affects the results of conventional fMRI-constrained EEG/MEG source imaging if no major mismatch between the two modalities is detected; while the new results become similar to those of typical EEG/MEG source imaging without fMRI constraint if the mismatch level is significant. A preliminary simulation study using realistic EEG signals demonstrated that the proposed technique can be a promising tool to selectively apply fMRI prior information to EEG/MEG source imaging. PMID- 17110767 TI - Imaging using parallel integrals in optical projection tomography. AB - We develop and demonstrate improved image-forming optics for optical projection tomography (OPT), with which the parallel integral throughout an object can be obtained. This method results in an improved resolution for OPT images, especially for the cross sections far from the optical axis of the image-forming optics. We find the optimal configuration used in our OPT system by use of a point spread function and simulation technique. The new method is validated by both numerical simulations and experimental results. The spatial resolution of the OPT system presented is approximately 40 microm. PMID- 17110768 TI - Dielectric measurement: error analysis and assessment of uncertainty. AB - The advantages and limitations of using partial differential analysis to assess the methodological uncertainty associated with the measurement of the dielectric properties of a material are discussed and an alternative pragmatic approach is proposed. It relies on repeat measurements of the dielectric properties of reference liquids and an analysis to estimate random and systematic uncertainties. Examples of measurement uncertainty are provided for well-defined monomolecular materials and for less homogeneous materials at microwave frequencies. All examples relate to measurement with an open-ended coaxial probe but the methodology is not specific to this technique. Examination of the components of uncertainty in the dielectric properties of biological tissue shows that, where the system is free of methodological bias, random fluctuations originating from sampling and natural inhomogeneity dominate the uncertainty budget. In such cases, the mean value of the measured parameter and the standard error of the mean can be taken as a good measure of the true value and its associated uncertainty. PMID- 17110769 TI - Morphological study of endothelial cells during freezing. AB - Microvascular injury is recognized as a major tissue damage mechanism of ablative cryosurgery. Endothelial cells lining the vessel wall are thought to be the initial target of freezing. However, details of this injury mechanism are not yet completely understood. In this study, ECMatrix 625 was used to mimic the tumour environment and to allow the endothelial cells cultured in vitro to form the tube like structure of the vasculature. The influence of water dehydration on the integrity of this structure was investigated. It was found that the initial cell shape change was mainly controlled by water dehydration, dependent on the cooling rate, resulting in the shrinkage of cells in the direction normal to the free surface. As the cooling was prolonged and temperature was lowered, further cell shape change could be induced by the chilling effects on intracellular proteins, and focal adhesions to the basement membrane. Quantitative analysis showed that the freezing induced dehydration greatly enhanced the cell surface stresses, especially in the axial direction. This could be one of the major causes of the final breaking of the cell junction and cell detachment. PMID- 17110770 TI - In vivo micro-CT lung imaging via a computer-controlled intermittent iso-pressure breath hold (IIBH) technique. AB - Respiratory research with mice using micro-computed tomography (micro-CT) has been predominantly hindered by the limited resolution and signal-to-noise ratio (SNR) as a result of respiratory motion artefacts. In this study, we develop a novel technique for capturing the lung microstructure in vivo using micro-CT, through a computer-controlled intermittent iso-pressure breath hold (IIBH), to reduce respiratory motion, increasing resolution and SNR of reconstructed images. We compare four gating techniques, i.e. no gating, late expiratory (LE) gating, late inspiratory (LI) gating and finally intermittent iso-pressure breath hold (IIBH) gating. Quantitatively, we compare several common image analysis methods used to extract valuable physiologic and anatomic information from the respiratory system, and show that the IIBH technique produces the most representative and repeatable results. PMID- 17110771 TI - Experimental evaluation of a MOSFET dosimeter for proton dose measurements. AB - The metal oxide semiconductor field-effect transistor (MOSFET) dosimeter has been widely studied for use as a dosimeter for patient dose verification. The major advantage of this detector is its size, which acts as a point dosimeter, and also its ease of use. The commercially available TN502RD MOSFET dosimeter manufactured by Thomson and Nielsen has never been used for proton dosimetry. Therefore we used the MOSFET dosimeter for the first time in proton dose measurements. In this study, the MOSFET dosimeter was irradiated with 190 MeV therapeutic proton beams. We experimentally evaluated dose reproducibility, linearity, fading effect, beam intensity dependence and angular dependence for the proton beam. Furthermore, the Bragg curve and spread-out Bragg peak were also measured and the linear-energy transfer (LET) dependence of the MOSFET response was investigated. Many characteristics of the MOSFET response for proton beams were the same as those for photon beams reported in previous papers. However, the angular MOSFET responses at 45, 90, 135, 225, 270 and 315 degrees for proton beams were over responses of about 15%, and moreover the MOSFET response depended strongly on the LET of the proton beam. This study showed that the angular dependence and LET dependence of the MOSFET response must be considered very carefully for quantitative proton dose evaluations. PMID- 17110772 TI - A nanodosimetry-based linear-quadratic model of cell survival for mixed-LET radiations. AB - A new nanodosimetry-based linear-quadratic (LQ) model of cell survival for mixed LET radiations has been developed. The new model employs three physical quantities and three biological quantities. The three physical quantities are related to energy depositions at two nanometre scales, 5 nm and 25 nm. The three biological quantities are related to the lesion production and interaction probabilities and the lesion repair rate. The coefficients alpha and beta of the LQ formula (alpha D + beta D(2)) are explicitly expressed in terms of the three physical quantities and the three biological quantities. The new model is shown to be consistent with the previously published cell survival curves of V-79 cells. The advantage of this new model is that it can be conveniently adopted to estimate the iso-effect for radiotherapies that involve ionizing radiation of mixed LET. An example is given to estimate the cell survival fractions for a high dose-rate mixed neutron and gamma-ray field from a (252)Cf source. PMID- 17110773 TI - Distributions of positron-emitting nuclei in proton and carbon-ion therapy studied with GEANT4. AB - Depth distributions of positron-emitting nuclei in PMMA phantoms are calculated within a Monte Carlo model for heavy-ion therapy (MCHIT) based on the GEANT4 toolkit (version 8.0). The calculated total production rates of (11)C, (10)C and (15)O nuclei are compared with experimental data and with corresponding results of the FLUKA and POSGEN codes. The distributions of e(+) annihilation points are obtained by simulating radioactive decay of unstable nuclei and transporting positrons in the surrounding medium. A finite spatial resolution of the positron emission tomography (PET) is taken into account in a simplified way. Depth distributions of beta(+)-activity as seen by a PET scanner are calculated and compared to available data for PMMA phantoms. The obtained beta(+)-activity profiles are in good agreement with PET data for proton and (12)C beams at energies suitable for particle therapy. The MCHIT capability to predict the beta(+)-activity and dose distributions in tissue-like materials of different chemical composition is demonstrated. PMID- 17110774 TI - Observations on the relationship between magnetic field characteristics and exposure conditions. AB - The authors have been investigating metrics of extremely low frequency magnetic field exposure under different circumstances. In this paper, we describe the properties of magnetic fields in homes, in the vicinity of powerlines, on trains and from a library security system. We conclude that there are key differences between each of these fields. This suggests that there may be a characterizable pattern for magnetic fields in different situations. PMID- 17110775 TI - The effect of covariance between the K alpha and the K beta lead peak concentrations on the uncertainty in the result of in vivo (109)Cd KXRF bone lead measurement. AB - The effect of covariance between the K alpha and K beta lead peak concentrations on the uncertainty in the (109)Cd K x-ray fluorescence measurement of lead in bone is addressed here. It is commonly believed that this covariance arises as a result of the mutual dependence of the ratios of x-ray to coherent amplitudes on the same coherent peak amplitude. Previous work assumes statistical independence between spectral quantities of interest, crudely estimates the uncertainties in the lead peak concentrations, and suggests that the effect of covariance on the measurement uncertainty is small and can be ignored. Consequently, the current method followed by most laboratories reports the measurement uncertainty as if the fluctuations in the measured peak concentrations were independent. The robustness of such assumption, however, is undermined by existing epidemiological data. This paper assesses the magnitude of the covariance effect, using a method based on the observed significant correlations between the ratios of x-ray to coherent peak amplitudes in series of repeat phantom measurements. The revised uncertainties following this approach can exceed the uncertainties estimated by the accepted method by as much as 40%, which suggests a much stronger effect of covariance on the measurement uncertainty than previously reported. PMID- 17110776 TI - Cell model for efficient simulation of wave propagation in human ventricular tissue under normal and pathological conditions. AB - In this paper, we formulate a model for human ventricular cells that is efficient enough for whole organ arrhythmia simulations yet detailed enough to capture the effects of cell level processes such as current blocks and channelopathies. The model is obtained from our detailed human ventricular cell model by using mathematical techniques to reduce the number of variables from 19 to nine. We carefully compare our full and reduced model at the single cell, cable and 2D tissue level and show that the reduced model has a very similar behaviour. Importantly, the new model correctly produces the effects of current blocks and channelopathies on AP and spiral wave behaviour, processes at the core of current day arrhythmia research. The new model is well over four times more efficient than the full model. We conclude that the new model can be used for efficient simulations of the effects of current changes on arrhythmias in the human heart. PMID- 17110777 TI - Quantification in simultaneous (99m)Tc/(123)I brain SPECT using generalized spectral factor analysis: a Monte Carlo study. AB - In SPECT, simultaneous (99m)Tc/(123)I acquisitions allow comparison of the distribution of two radiotracers in the same physiological state, without any image misregistration, but images can be severely distorted due to cross-talk between the two isotopes. We propose a generalized spectral factor analysis (GSFA) method for solving the cross-talk issue in simultaneous (99m)Tc/(123)I SPECT. In GSFA, the energy spectrum of the photons in any pixel is expressed as a linear combination of five common spectra: (99m)Tc and (123)I photopeaks and three scatter spectra. These basis spectra are estimated from a factor analysis of all spectra using physical priors (e.g. Klein-Nishina distributions). GSFA was evaluated on (99m)Tc/(123)I Monte Carlo simulated data and compared to images obtained using recommended spectral windows (WIN) and to the gold standard (GS) images (scatter-free, cross-talk-free and noise-free). Using GSFA, activity concentration differed by less than 9% compared to GS values against differences from -23% to 110% with WIN in the (123)I and (99m)Tc images respectively. Using GSFA, simultaneous (99m)Tc/(123)I imaging can yield images of similar quantitative accuracy as when using sequential and scatter-free (99m)Tc/(123)I imaging in brain SPECT. PMID- 17110778 TI - Methods and evaluations of MRI content-adaptive finite element mesh generation for bioelectromagnetic problems. AB - In studying bioelectromagnetic problems, finite element analysis (FEA) offers several advantages over conventional methods such as the boundary element method. It allows truly volumetric analysis and incorporation of material properties such as anisotropic conductivity. For FEA, mesh generation is the first critical requirement and there exist many different approaches. However, conventional approaches offered by commercial packages and various algorithms do not generate content-adaptive meshes (cMeshes), resulting in numerous nodes and elements in modelling the conducting domain, and thereby increasing computational load and demand. In this work, we present efficient content-adaptive mesh generation schemes for complex biological volumes of MR images. The presented methodology is fully automatic and generates FE meshes that are adaptive to the geometrical contents of MR images, allowing optimal representation of conducting domain for FEA. We have also evaluated the effect of cMeshes on FEA in three dimensions by comparing the forward solutions from various cMesh head models to the solutions from the reference FE head model in which fine and equidistant FEs constitute the model. The results show that there is a significant gain in computation time with minor loss in numerical accuracy. We believe that cMeshes should be useful in the FEA of bioelectromagnetic problems. PMID- 17110779 TI - Prediction of mechanical properties of trabecular bone using quantitative MRI. AB - Techniques for quantitative magnetic resonance imaging (MRI) have been developed for non-invasive estimation of the mineral density and structure of trabecular bone. The R*(2) relaxation rate (i.e. 1/T*(2)) is sensitive to bone mineral density (BMD) via susceptibility differences between trabeculae and bone marrow, and by binarizing MRI images, structural variables, such as apparent bone volume fraction, can be assessed. In the present study, trabecular bone samples of human patellae were investigated in vitro at 1.5 T to determine the ability of MRI derived variables (R*(2) and bone volume fraction) to predict the mechanical properties (Young's modulus, yield stress and ultimate strength). Further, the MRI variables were correlated with reference measurements of volumetric BMD and bone area fraction as determined with a clinical pQCT system. The MRI variables correlated significantly (p < 0.01) with the mechanical variables (r = 0.32 0.46), BMD (r = 0.56) and bone structure (r = 0.51). A combination of R*(2) and MRI-derived bone volume fraction further improved the prediction of yield stress and ultimate strength. Although pQCT showed a trend towards better prediction of the mechanical properties, current results demonstrate the feasibility of combined MR imaging of marrow susceptibility and bone volume fraction in predicting the mechanical strength of trabecular bone and bone mineral density. PMID- 17110780 TI - An analytical algorithm for skew-slit collimator SPECT with uniform attenuation correction. AB - To replace the conventional pinhole (normal cone-beam) collimator, a novel skew slit collimator was previously proposed and a Novikov-type algorithm developed to reconstruct images using the skew-slit geometry. The goal of this paper is to develop a reconstruction algorithm that has better noise control than the Novikov type algorithm. The new algorithm is able to compensate for uniform attenuation, and computer simulation results show that reconstructed images are less noisy. PMID- 17110781 TI - Analysis of the electric field induced forces in erythrocyte membrane pores using a realistic cell model. AB - We calculate the induced electric stress forces on transient hydrophobic pores in the membrane of an erythrocyte exposed to an electric field. For this purpose, we use a finite element numerical technique and a realistic shape for the biconcave erythrocyte represented by a set of parametric equations in terms of Jacobi elliptic functions. The results clearly show that the electrical forces on the base and sidewalls of the pore favour the opening of the pore. A comparison of the force densities obtained for an unstretched flat membrane and for the realistic erythrocyte model shows that the thinning and curvature of the membrane cannot be neglected. We also show that the pore deformation depends strongly on the orientation of the pore with respect to the external field, and in particular is very small when the field is tangent to the membrane surface. PMID- 17110782 TI - Small nodule detectability evaluation using a generalized scan-statistic model. AB - In this paper is investigated the use of the scan statistic for evaluating the detectability of small nodules in medical images. The scan-statistic method is often used in applications in which random fields must be searched for abnormal local features. Several results of the detection with localization theory are reviewed and a generalization is presented using the noise nodule distribution obtained by scanning arbitrary areas. One benefit of the noise nodule model is that it enables determination of the scan-statistic distribution by using only a few image samples in a way suitable both for simulation and experimental setups. Also, based on the noise nodule model, the case of multiple targets per image is addressed and an image abnormality test using the likelihood ratio and an alternative test using multiple decision thresholds are derived. The results obtained reveal that in the case of low contrast nodules or multiple nodules the usual test strategy based on a single decision threshold underperforms compared with the alternative tests. That is a consequence of the fact that not only the contrast or the size, but also the number of suspicious nodules is a clue indicating the image abnormality. In the case of the likelihood ratio test, the multiple clues are unified in a single decision variable. Other tests that process multiple clues differently do not necessarily produce a unique ROC curve, as shown in examples using a test involving two decision thresholds. We present examples with two-dimensional time-of-flight (TOF) and non-TOF PET image sets analysed using the scan statistic for different search areas, as well as the fixed position observer. PMID- 17110783 TI - Pencil kernel correction and residual error estimation for quality-index-based dose calculations. AB - Experimental data from 593 photon beams were used to quantify the errors in dose calculations using a previously published pencil kernel model. A correction of the kernel was derived in order to remove the observed systematic errors. The remaining residual error for individual beams was modelled through uncertainty associated with the kernel model. The methods were tested against an independent set of measurements. No significant systematic error was observed in the calculations using the derived correction of the kernel and the remaining random errors were found to be adequately predicted by the proposed method. PMID- 17110784 TI - Mechanical strength of sarcomere structures of skeletal myofibrils studied by submicromanipulation. AB - The mechanical strength of sarcomere structures of skeletal muscle was studied by rupturing single myofibrils of rabbit psoas muscle by submicromanipulation techniques. Microbeads coated with alpha-actinin were attached to the surface of myofibrils immobilized to coverslip. By use of either optical tweezers or atomic force microscope, the attached beads were captured and detached from the myofibrils. During the detachment of the beads, the actin filaments bound specifically to the beads were peeled off from the bulk structures of myofibrils, thus rupturing the peripheral components of the myofibrils bound to the actin filaments. By analyzing the ruptures thus produced in various myofibril preparations, it was found that the sarcomere structure of myofibrils is maintained by numerous molecular components having the mechanical strength sufficient to sustain the contractile force produced by the actomyosin system. The present techniques could be applied to study the mechanical strength of cellular organelles containing actin filaments as their component. PMID- 17110785 TI - XBP1 is critical to protect cells from endoplasmic reticulum stress: evidence from Site-2 protease-deficient Chinese hamster ovary cells. AB - XBP1 is a transcription factor downstream of IRE1, a transmembrane protein in the endoplasmic reticulum (ER) which functions as a sensor and transducer of ER stress. XBP1 mRNA is constitutively expressed at a low level as an intron containing precursor mRNA (unspliced mRNA), which is subject to IRE1-mediated splicing reaction upon ER stress to produce the active form of XBP1, pXBP1(S). Because the XBP1 promoter carries a perfect ER stress-response element, namely, the cis-acting element responsible for the induction of ER chaperones, and XBP1 mRNA is induced in response to ER stress with a time course similar to that of ER chaperone mRNAs, it is conjectured that transcription factor ATF6, activated immediately upon ER stress, induces the transcription of not only ER chaperone genes but also of XBP1 gene, such that pXBP1(S) produced by the splicing of an increased level of XBP1 mRNA escapes from proteasome-mediated degradation. Here, we examined this notion by determining the induction of XBP1 mRNA and pXBP1(S) in mutant Chinese hamster ovary (M19) cells deficient in Site-2 protease, which executes the last step of ER stress-induced activation of ATF6. We found that the induction of XBP1 mRNA and pXBP1(S) was greatly reduced in M19 cells as compared with wild-type cells, leading to a marked reduction in the extent of induction of XBP1-target gene. M19 cells were much more sensitive to ER stress than wild-type cells. Importantly, overexpression of XBP1 unspliced mRNA in M19 cells reversed all of these phenotypes. We concluded that ATF6-mediated induction of XBP1 mRNA is important to the production of pXBP1(S), activation of XBP1-target genes, and protection of cells from ER stress. PMID- 17110786 TI - Analysis of ATF6 activation in Site-2 protease-deficient Chinese hamster ovary cells. AB - Mammalian transcription factor ATF6 is constitutively synthesized as a type II transmembrane protein embedded in the endoplasmic reticulum (ER). It is activated when unfolded proteins are accumulated in the ER under ER stress through a process called regulated intramembrane proteolysis (Rip), in which ATF6 is transported from the ER to the Golgi apparatus where it undergoes sequential cleavage by Site-1 and Site-2 proteases. The cytosolic transcription factor domain of ATF6 liberated from the Golgi membrane enters the nucleus where it activates transcription of ER-localized molecular chaperones and folding enzymes, leading to the maintenance of the homeostasis of the ER. Here, we analyzed M19 cells, a mutant of Chinese hamster ovary cells deficient in Site-2 protease. It was previously shown that M19 cells are defective in the induction of mRNA encoding the major ER chaperone BiP. In M19 cells, ATF6 was not converted from the membrane-bound precursor form to the cleaved and nuclear form as expected. Moreover, some of the ATF6 was constitutively relocated to the Golgi apparatus, where it was cleaved by Site-1 protease, and remained associated with the Golgi apparatus, indicating that the ER of M19 cells was constitutively stressed. Consistent with this notion, the two other ER stress response mediators, IRE1 and PERK, were also constitutively activated in M19 cells. M19 cells showed inefficient secretion of a model protein. These results suggest that Rip-mediated activation of ATF6 is important for the homeostasis of the ER in not only ER stressed but also unstressed cells. PMID- 17110787 TI - The use of ionizers to destroy allergens: past, present and future research. PMID- 17110788 TI - TSA-induced cell death in prostate cancer cell lines is caspase-2 dependent and involves the PIDDosome. AB - The histone deacetylase inhibitor Trichostatin A (TSA) has previously been found to induce caspase activity in the human prostate cancer cell lines DU145 and LNCaP. TSA treatment resulted in the release of cytochrome c and Smac/DIABLO from mitochondria in DU145, and activation of caspase-9 in both cell lines. We concluded that TSA mediated its effect via the mitochondrial pathway. The aim of the current study was to determine how TSA initiated the caspase cascade. The results revealed that caspase-2 plays an important role in TSA-induced apoptosis. Inhibition of caspase-2 by siRNA or expression of caspase-2dn substantially decreased caspase activity after TSA treatment in both cell lines, siRNA caspase 2 also inhibited TSA-induced cell death. Caspase-2 acts upstream of caspase-8 and -9 and mediates mitochondrial cytochrome c release. Coimmunoprecipitation experiments show that caspase-2 formed protein complexes with RADD/RAIDD and PIDD. Together, these data indicate that caspase-2 initiates caspase cascade after TSA treatment and involves the formation of the PIDDosome. PMID- 17110791 TI - Delayed effects of antidepressant drugs in rats. AB - The present study has addressed the question of what is more important for the occurrence of adaptive changes observed in the organism treated with antidepressant drugs: a daily dosing of the drug or the period of time necessary for the plastic events to develop. Here, we report on the effects of desipramine given to rats acutely (and tested following 2 drug-free weeks) as when the drug was administered repeatedly, on behavior in the forced swim test (i.e. significant shortening of immobility time by ca. 60%) and on the binding of [3H]CGP12177 to beta-adrenergic receptors in the rat brain cortex (significant decrease of the binding by ca. 15%). Additionally, using the procedure of the repeated forced swim test (six times over 21 days), we show that the shortening of immobility time induced by a single dose of imipramine persisted throughout the whole experimental period and was similar to that seen in a group of animals treated repeatedly with the drug. Also, the effects of citalopram on immobility and climbing were similar after acute treatment and delayed testing to those seen after repeated drug exposure. The results obtained in the present study may question some conclusions that are usually drawn from the behavioral and, especially, biochemical studies concerning the need for repeated treatment with antidepressant drugs to induce various adaptive changes in the brain, which are thought to be responsible for the therapeutic efficacy of these drugs. PMID- 17110792 TI - A review of delay-discounting research with humans: relations to drug use and gambling. AB - Delay discounting represents the extent to which consequences, or outcomes, decrease in effectiveness to control behavior as a function of there being a delay to their occurrence. Higher rates of delay discounting are often operationalized as an index of impulsivity, and as such impulsive discounting may hold considerable potential for understanding fundamental behavioral processes associated with a range of problematic behaviors - including drug use and pathological gambling. This paper first provides a review of several assessment methods used in delay-discounting research with humans. Following, the delay discounting literature related to drug use and gambling is reviewed. Consistencies across this literature are identified; and future research directions are discussed, which include (a) improving methods of assessment for delay discounting and (b) moving drug-use research progressively to causal interpretations, with high rates of delay discounting either predisposing to drug use or resulting from drug use itself. PMID- 17110793 TI - Delay discounting in college cigarette chippers. AB - Individuals who smoke cigarettes regularly but do not become dependent on them provide a unique opportunity for studying the factors that inhibit drug dependence. Previous research on this population, sometimes referred to as 'cigarette chippers', showed that they did not differ from regular smokers in terms of smoking topography (e.g. puff number and duration) and circulating nicotine levels, but that they did show more self-control according to answers on a questionnaire. We evaluated the generality of this finding using a behavioral choice procedure. The participants were undergraduate students (n=71), who were regular smokers, chippers, or nonsmokers. In the choice procedure, one option was a smaller but sooner amount of money, and the other option was a larger but delayed amount of money. Under these conditions, preference for the sooner smaller amount implies that the later larger monetary amounts were discounted. It is widely assumed that the rate of discounting provides an operational definition of impulsivity. In one version of the procedure, the money was hypothetical. In a second version, each choice had a chance of producing an actual monetary outcome. When there was an actual monetary outcome, regular smokers were more likely to choose the sooner but smaller monetary option than chippers and nonsmokers. For all participants, the rate of discounting decreased as the magnitude of the monetary outcomes increased, and for smokers and chippers the differences in discount rates in the two versions of the delayed outcome procedure were the same. These findings are consistent with the view that chippers are less impulsive than smokers. Quantitative aspects of these findings led to the hypothesis that discount rates decrease as a negative power function of the monetary value of the options. This result establishes an analogy between delay discounting experiments and psychophysical experiments. Results from two earlier studies support the analogy. PMID- 17110794 TI - Modeling the anxiety-depression continuum hypothesis in domestic fowl chicks. AB - Anxiety and depression are currently classified as separate clinical syndromes despite considerable similarities in their symptoms, pathophysiological substrates and response to treatment interventions. An alternative hypothesis views anxiety and depression along a temporal continuum, a construct that the current research attempts to model in a preclinical setting. In experiment 1, socially raised domestic fowl chicks separated from conspecifics demonstrated a pattern of distress vocalizations that sequentially models anxiety-like and depressive-like states. In addition, administration of the benzodiazepine anxiolytic chlordiazepoxide and the tricyclic antidepressant imipramine provided pharmacological validation for the model in that they were capable of dissociating the anxiety-like and depressive-like states. In experiment 2, corticosterone levels were quantified across the isolation test session to provide convergent validity to the model. These findings fit well with the human clinical literature on the anxiety-depression continuum perspective, and suggest the consideration of a nosology that emphasizes the inter-relatedness of these clinical states rather than their boundaries. PMID- 17110795 TI - Differential effects of micro-opioid, delta-opioid and kappa-opioid receptor agonists on dopamine receptor agonist-induced climbing behavior in mice. AB - Interactions between the dopaminergic system and opioids have not been adequately clarified. The present study was designed to investigate the effects of micro opioid (morphine), delta-opioid (SNC80) and kappa-opioid (U50 488H) receptor agonists on dopamine receptor agonist-induced climbing behavior in mice. Apomorphine (dopamine-receptor agonist) increased stereotyped climbing behavior, unlike methamphetamine, morphine, U-50 488H and (+/-)7-hydroxy-N,N-di-n-propyl-2 aminotetralin hydrobromide (D2-like receptor agonist). Furthermore, SKF81297 (D1 receptor agonist) and SNC80 caused climbing behavior. In addition, while morphine (20 mg/kg), but not U50 488H or SNC80, significantly attenuated high-dose apomorphine (2.0 mg/kg)-induced climbing behavior, it significantly potentiated low-dose apomorphine (0.5 mg/kg)-induced climbing behavior. These results suggest that morphine may have dual effects on the behavioral effects induced by apomorphine. Furthermore, we interestingly showed that the combination of apomorphine or SKF81297 and SNC80 enhanced frequent nonstereotypic climbing behavior, suggesting that delta/D1 interactions may play a prominent role in the expression of certain types of behavior in mice. Thus, micro-opioid, delta-opioid and kappa-opioid receptor agonists induce possible differential effects on the dopaminergic system in mice. PMID- 17110796 TI - Antidepressant-like effects of agomelatine (S 20098) in the learned helplessness model. AB - To confirm the antidepressant-like activity of agomelatine (S 20098), a melatonin agonist and 5-hydroxytryptamine2C antagonist, already reported in the chronic mild stress and forced swimming tests, the effects of agomelatine were investigated in the learned helplessness test and compared with those of imipramine, melatonin and a selective 5-hydroxytryptamine2C antagonist, SB-242 084. Agomelatine was administered for 5 days either once a day or twice a day, and the effects of pretreatment by a melatonin receptor antagonist, S 22153 (20 mg/kg/day), were studied. A deficit in avoidance learning was observed in helpless control animals. Agomelatine (10 mg/kg/day) administered once a day significantly reduced this deficit with an effect similar to that of imipramine. Effects of agomelatine were abolished by S 22153 pretreatment. Melatonin or SB 242 084 did not reduce the deficit of helpless control animals. These results confirm the antidepressant-like activity of agomelatine and suggest a role of melatonin receptors in its mechanism of action. PMID- 17110797 TI - Maternal deprivation specifically enhances vulnerability to opiate dependence. AB - Maternal deprivation has been shown to increase vulnerability to morphine dependence and to disturb the enkephalinergic system in adulthood. To study whether or not this vulnerability to opiates is a specific feature, we examined oral self-administration behaviour of various reinforcing substances. Experiments were performed with morphine (25 mg/l), ethanol (10%), amphetamine (25 mg/l) and cocaine (100 mg/l). Drugs were available in a continuous free choice paradigm during 3 months. Cocaine and ethanol consumption and preference were similar in both deprived and control rats. Deprived rats greatly increased their morphine consumption and 78% of them showed a progressive decrease in morphine aversion. Only a slight, but significant, increase in oral amphetamine consumption was observed in deprived rats when compared with control rats. The difference in amphetamine self-administration in control and deprived rats cannot be explained by a modification of dopamine transporter expression measured by immunoautoradiography. Altogether, we conclude that maternal deprivation worsens inherent susceptibility to dependence, specifically for opiates, and therefore represents a highly valuable model to study environmentally triggered interindividual vulnerability to opiate addiction. PMID- 17110798 TI - Measuring forelimb force control and movement in Fischer 344/Brown Norway rats: effects of age and lorazepam. AB - The purpose of this study was to measure forelimb force control and movement kinetics in rats, as they are affected by normal aging and the benzodiazepine lorazepam. Young (6 months), middle-aged (18 months), and aged (24 months) rats were trained to emit discrete forelimb responses on an isometric force disk within a 20-25 g force band for water reinforcement. Dependent variables included number of responses, percentage of reinforced responses, peak response forces, and inter-response times. Inter-response times were divided into two categories: inter-response times <0.5 s (reflecting rapid, discrete forelimb responses) and inter-response times 4-8 s (reflecting movement sequences). Aged rats exhibited no apparent deficits in forelimb force control. Although older rats emitted fewer responses than younger rats, their response accuracy was greater. Peak forces did not differ among the groups. Both categories of inter-response times were slower in the aged group, reflecting slowed discrete movements and movement sequencing. Lorazepam increased the number of responses and peak forces, decreased response accuracy, and lengthened inter-response times within the 4-8 s range (but not the <0.5 s range) in all age groups. The results suggest that movement sequences may be more sensitive to the effects of acute benzodiazepines than rapid discrete movements. PMID- 17110799 TI - Adrenergic blocker carvedilol attenuates the cardiovascular and aversive effects of nicotine in abstinent smokers. AB - The cardiovascular response to nicotine is mediated mainly by noradrenergic activation. Whether noradrenergic activation mediates other effects of nicotine has not been well documented in humans. In this study, we examined the effects of an alpha and beta-adrenergic receptor blocker: carvedilol, on cardiovascular and subjective responses to nicotine lozenge and on the ability of nicotine lozenge to suppress tobacco withdrawal symptoms in overnight abstinent smokers. Fifteen smokers, nine men and six women, participated in a double-blind, placebo controlled, crossover study. In each of the three experimental sessions, participants were treated orally with a single 25 or 50 mg dose of carvedilol or placebo. Two hours and 10 min following the medication treatment, participants received a single 4 mg nicotine lozenge. Carvedilol treatment attenuated the nicotine-induced heart rate, systolic and diastolic blood pressure increases. Carvedilol also attenuated the self-report rating of 'bad effects' in response to nicotine. Carvedilol, alone or in combination with nicotine lozenge, did not affect tobacco withdrawal symptoms. Carvedilol treatment did not affect performance on the Stroop Test. These results support the effectiveness of carvedilol for attenuating the cardiovascular effects of nicotine. Attenuation of the rating of 'bad effects' by carvedilol suggests that noradrenergic activation may also mediate the aversive effects of nicotine. PMID- 17110800 TI - The effect of flutamide on systemic and renal hemodynamics in Zucker diabetic rats: paradoxic renal vasodilator response to endothelin-1 and TXA2 receptor activation in female sex. AB - BACKGROUND: There is increasing evidence that endogenous sex hormones regulate vascular reactivity, and testosterone may contribute to the worse prognosis for renal disease in men. Male Zucker diabetic rats exhibit improved renal hemodynamic responses after castration. It is, however, unclear whether endogenous testosterone affects renal and systemic microcirculatory responses in the female sex, especially in type 2 diabetes. AIM: To test the hypothesis that endogenous testosterone in the female Zucker diabetic rat exerts a pathophysiologically relevant modulation of endothelial and renal microvascular function. METHODS: Female Zucker diabetic rats (FZDR) aged 5-6 weeks and from the same litter were divided into 2 groups (n = 6-8 each). The experimental group received the androgen receptor blocker flutamide, dissolved in alcohol and added to their drinking water (500 mL) at 20 mg/rat/week. The control FZDR received only the alcohol vehicle added to the same volume of drinking water. Both FZDR groups were treated for 3 months before undergoing the hemodynamic studies. A sex comparison control group of male Zucker diabetic rats (MZDR), also aged 5-6 weeks, was studied, following same protocol. Mean arterial pressure (MAP) and renal cortical blood flow (RCF) response to phenylephrine, acetylcholine, TXA2 mimetic U46619, endothelin-1 (ET-1), angiotensin II, and L-NG-nitro arginine methyl ester were studied. Furthermore, the role of protein kinase C in the responses was assessed using phorbol-12,13 dibutyrate 10(-4) M. The impact of flutamide on body weights and blood glucose of the rats were also determined. RESULTS: Flutamide-treated FZDR had a significant reduction in body weight/adiposity to 432 +/- 44 g, compared to controls at 553 +/- 37 g (P = 0.045), and random blood glucose concentration of 185 +/- 44 g/dL, compared to the control FZDR at 475 +/- 34 g/dL (P = 0.002). Vehicle-treated FZDR (n = 6-8), exhibited little or no systemic or renal response to any of the agonists. By contrast, flutamide treatment of FZDR (n = 5-7) caused a normalization of the dose-dependent MAP and RCF pressor response to phenylephrine [P < 0.005, analysis of variance (ANOVA)] and the vasodilator response to acetylcholine (P <. 0.01, ANOVA). Flutamide-treated FZDR showed enhanced pressor response to U46619 (P = 0.024, ANOVA), ET-1, and angiotensin II (P < 0.03, ANOVA). Surprisingly, the augmented systemic pressor action of U46619 and ET-1 was accompanied by a renal vasodilator action, with paradoxic RCF increases to U46619 (P < 0.003, ANOVA) and to ET-1 (P < 0.001, ANOVA) only in flutamide-treated FZDR. By contrast, flutamide treated MZDR exhibited no significant change in body weight and an attenuation of the vasoconstrictor responses and enhanced nitric oxide-mediated dilatation compared with male controls. However, no specific effect on ET-1 or TXA2 receptor mediated renal perfusion was discernible. Both L-NG-nitro arginine methyl ester and the protein kinase C agonist phorbol-12,13 dibutyrate [10(-4)M] significantly increased MAP and reduced RCF (P < 0.03) in the experimental FZDR compared with their controls. CONCLUSION: Flutamide administration to FZDR resulted in the reversal of abnormal systemic and renal alpha-1-mediated vasoconstriction and enhanced nitric oxide-mediated vasodilation. Flutamide caused a paradoxic but specific increase in renal perfusion during ET-1 and TXA2 receptor activation, which could be renoprotective in females. The salutary effects of flutamide on vascular reactivity in the FZDR may be mediated by a protein kinase C-dependent mechanism. These results are compatible with the notion that endogenous testosterone may regulate systemic and renal microcirculation in the female sex and in the type 2 diabetic state. PMID- 17110801 TI - In vitro preclinical cardiac assessment of tolterodine and terodiline: multiple factors predict the clinical experience. AB - Terodiline and tolterodine are drugs used to treat urinary incontinence. Terodiline was removed from the market in 1991 for proarrhythmia, whereas tolterodine has a generally benign clinical cardiac profile. To assess differences in the electrophysiologic actions of these drugs, we evaluated their effects on hERG current (HEK cells) and cardiac Purkinje fiber repolarization. The IC50 for hERG block (37 degrees C) by tolterodine was 9.6 nM and by terodiline was 375 nM, values near or below clinical concentrations. Tolterodine elicited concentration-dependent prolongation of the action potential duration (APD90). In contrast, terodiline depressed the action potential plateau and induced triangulation without affecting APD90. The triangulation ratios (normalized ratio of APD50 over APD90) for terodiline were 0.94 and 0.59 for 1.0 and 10 microM and for tolterodine, were 0.99 and 0.97 at 7 and 70 nM. In summary, tolterodine, a potent hERG blocker, has a benign clinical cardiac profile at therapeutic concentrations that may be due to its lack of triangulation, as well as extensive plasma protein binding. However, at supratherapeutic concentrations, preclinical data predict risk of QT prolongation. These data suggest that hERG block and triangulation are among multiple factors that must be considered in preclinical cardiac safety assessments. PMID- 17110802 TI - Angiotensin-converting enzyme inhibitors influence left ventricular mass and function independently of the antihypertensive effect. AB - In our retrospective study, we evaluated whether ACE inhibitors can influence left ventricular (LV) morphofunctional characteristics in essential hypertension independently of the antihypertensive effect. We studied 21 hypertensive patients (group 1) before and after at least 18 months of treatment with ACE inhibitors that did not induce any blood pressure (BP) reduction; as a control group, we evaluated 19 hypertensive patients (group 2) not treated with antihypertensive drugs during the same period. At baseline, the 2 groups, neither one previously treated with antihypertensive drugs, were not significantly different with regard to sex, age, body mass index, 24-hour BP, and heart rate; LV mass index was similar between the groups, whereas LV diastolic indices were significantly lower in group 1. At the second evaluation, body mass index, 24-hour BP, and heart rate were unchanged in both groups; LV mass index was significantly decreased in group 1 and increased in group 2. LV diastolic parameters were significantly improved in group 1, whereas in group 2, diastolic function was significantly deteriorated. In conclusion, our clinical study shows that ACE inhibitors can induce LV hypertrophy regression and improvement of diastolic function also in the absence of any antihypertensive effect. PMID- 17110803 TI - Administration of pioglitazone in low-density lipoprotein receptor-deficient mice inhibits lesion progression and matrix metalloproteinase expression in advanced atherosclerotic plaques. AB - Recent clinical trials have provided evidence that pioglitazone reduces cardiovascular events in patients with type 2 diabetes. However, the underlying mechanisms are not well understood. Because it has been well established that disruption of atherosclerotic plaques is a key event involved in acute myocardial infarction, we hypothesized that pioglitazone reduces cardiovascular events by stabilizing atherosclerotic lesions. In this study, we used an animal model to test our hypothesis. Low-density lipoprotein receptor-deficient (LDLR-/-) male mice were first fed a high-fat diet for 4 months to induce the formation of aortic atherosclerotic plaques and then treated with pioglitazone for the next 3 months. Analysis of atherosclerotic plaques at the end of the study showed that treatment with pioglitazone at 20 mg/kg/day reduced the progression of atherosclerotic plaques as compared to untreated mice. Furthermore, gene array analysis, quantitative real-time polymerase chain reaction, and immunohistochemical analysis showed that pioglitazone inhibited high-fat diet induced upregulation of matrix metalloproteinase (MMP) expression. Finally, Sirius red staining showed that atherosclerotic lesions in mice receiving pioglitazone had higher collagen contents than those in untreated mice. This study demonstrated for the first time that administration of pioglitazone in LDLR /- mice inhibited lesion progression and MMP expression in established atherosclerotic plaques and thus delineated a potential mechanism by which pioglitazone reduces cardiovascular events in patients with type 2 diabetes. PMID- 17110804 TI - Chronic direct stimulation of adenylyl cyclase induces cardiac desensitization to catecholamine and beta-adrenergic receptor downregulation in rabbits. AB - Chronic stimulation of beta-adrenergic receptors (betaARs) induces betaAR downregulation. However, it is not known whether continuous activation of adenylyl cyclase without direct stimulation of betaARs leads to receptor downregulation. This study investigated the effects of chronic stimulation of adenylyl cyclase with colforsin, on hemodynamic variables, and on myocardial betaAR density. In all, 55 rabbits received intravenous colforsin (1.6 microg/kg/min, n = 20), isoproterenol (ISO; 0.4 microg/kg/min, n = 16), or saline (n = 19) for two weeks. After chronic drug administration, responses of systolic (Delta% peak LV +dP/dt) and diastolic function (Delta% peak LV -dP/dt), and heart rate (Delta% heart rate), to acute administration of ISO (0.05 to 0.2 microg/kg/min) or colforsin (5 to 20 nmol/kg/min) were decreased compared to those before chronic administration. betaAR density in the colforsin group (69.8 +/- 13.8 fmol/ml protein) was less than that in the saline group (79.8 +/- 15.0 fmol/ml protein, P < 0.05), but was greater than that in the ISO group (56.3 +/- 8.4 fmol/ml protein, P < 0.05). Thus, chronic direct stimulation of adenylyl cyclase elicited systolic and diastolic functional desensitization to betaAR stimulation or adenylyl cyclase stimulation, and myocardial betaAR downregulation. PMID- 17110805 TI - Treatment with raloxifene and 17beta-estradiol differentially modulates nitric oxide and prostanoids in venous endothelium and platelets of ovariectomized pigs. AB - Oral treatment with raloxifene, a synthetic estrogen receptor modulator (SERM), or 17beta-estradiol (E2) increases risk for venous thrombosis in women. Acute application of either substance releases endothelium-derived factors from isolated femoral veins but it is not known how their chronic use affects venous functions or the interaction of platelets with veins. This study tested the hypothesis that treatment of ovariectomized animals with oral raloxifene or E2 would increase release of proaggregatory factors from venous endothelium and platelets. Ovariectomized (OVX) pigs were either untreated or treated with oral raloxifene (60 mg/day) or E2 (2 mg/day) for 4 weeks. Plasma concentrations of nitric oxide were comparable in both treatment groups and greater than in OVX pigs. Ratio of plasma thromboxane to prostacyclin was twofold greater in raloxifene compared to E2-treated pigs. In isolated femoral veins, NG-monomethyl L-arginine (L-NMMA; 10(-4) M) augmented endothelium-dependent relaxations to adenosine diphosphate in veins from E2-treated pigs but inhibited relaxations in veins from raloxifene-treated pigs. Addition of indomethacin (10(-5) M) reversed these effects. Endothelium-dependent relaxations to thrombin were inhibited by L NMMA only in OVX and raloxifene-treated pigs. Autologous platelets contracted veins in all groups; the magnitude of contractions depended upon the number of platelets and existing tone. Basal release of thromboxane from platelets was greatest in raloxifene compared to OVX or E2-treated pigs. Raloxifene treatment compared to E2 increased production of contractile and proaggregatory prostanoids from venous endothelium and platelets. These differences, if found in humans, may contribute to varying degrees of thrombotic risk with the SERM compared to the natural hormone. PMID- 17110806 TI - Preclinical evaluation of S18886 in an experimental model of coronary arterial thrombosis. AB - The specific thromboxane receptor antagonist, S18886, was evaluated for prevention of coronary arterial thrombosis and myocardial ischemia-reperfusion in anesthetized canines. For the primary thrombosis study in left circumflex (LCX) coronary artery, 26 dogs were randomized to receive either vehicle (n = 7) or intravenous S18886 (0.3 mg/kg, n = 6; 1.0 mg/kg, n = 6; and 3.0 mg/kg, n = 7). The respective times to occlusion after S18886 were as follows: 56.8 +/- 9.3, 83.5 +/- 14.9, and 92.4 +/- 15.7 minutes compared to 43.3 +/- 8.2 minutes after vehicle. S18886 caused a minimal increase in tongue bleeding time and a significant decrease in ex vivo platelet aggregation to arachidonic acid or U46619. Another 37 dogs were randomized to receive placebo (n = 12), clopidogrel 1.0 mg/kg p.o. QDX3 (n = 9), clopidogrel + S18886 0.3 (n = 9) or 1.0 (n = 7) mg/kg intravenous. Clopidogrel produced a 50% reduction in adenosine diphosphate induced platelet aggregation and a slight increase in the time to occlusion. However, clopidogrel + S18886 1.0 mg/kg prevented occlusive thrombus formation in most of the coronary vessels over 6 hours. S18886 did not alter myocardial infarct size in the ischemia-reperfusion model. In conclusion, S18886 alone caused a dose-dependent prolongation in the time to primary occlusive coronary artery thrombosis, whereas S18886 + clopidogrel displayed effective in preventing occlusive thrombus formation with only a moderate increase of tongue-bleeding time. PMID- 17110807 TI - Treprostinil in advanced experimental pulmonary hypertension: beneficial outcome without reversed pulmonary vascular remodeling. AB - INTRODUCTION: Beneficial effects of treprostinil, a stable prostacyclin analogue, were demonstrated in patients with pulmonary arterial hypertension (PAH). Although regression of pulmonary vascular remodeling has been suggested as therapeutic mechanism, its mode of action remains unknown. METHODS: Flow associated PAH was created in rats by injection of monocrotaline (60 mg/kg) combined with an abdominal aortocaval shunt. Subsequently, rats were treated with subcutaneous treprostinil (50 ng/kg/min, treated; n = 8) or saline (untreated; n = 9). A control group underwent sham-surgery (n = 8). Animals were sacrificed at symptoms of cardiac failure, together with their matched controls. RESULTS: Dyspnea and weight loss determined the moment of sacrifice in 8/9 untreated animals (89%) versus in one of eight treated animals (13%; log-rank test survival curves; P = 0.02). Mean pulmonary arterial pressure increased in the model (42 +/ 2 mm Hg in untreated vs. 18 +/- 1 in controls; P < 0.01) and decreased by 8 mm Hg after therapy (34 +/- 3 mm Hg, P = 0.04 vs. untreated). No effects of treatment on right ventricular hypertrophy could be demonstrated. Quantitative morphometry of pre- and intra-acinar pulmonary arteries revealed no effects of treatment on vessel histopathology. CONCLUSIONS: Treprostinil treatment improved clinical course and ameliorated symptoms of heart failure in a model of advanced PAH. However, beneficial effects were not associated with reversed structural remodelling of the pulmonary vasculature. PMID- 17110808 TI - Acute effects of febuxostat, a nonpurine selective inhibitor of xanthine oxidase, in pacing induced heart failure. AB - We investigated whether xanthine oxidase inhibition with febuxostat enhances left ventricular (LV) function and improves myocardial high energy phosphates (HEP) in dogs with pacing-induced heart failure (CHF). Febuxostat (2.2 mg/kg over 10 minutes followed by 0.06 mg/kg/min) caused no change of LV function or myocardial oxygen consumption (MVO2) at rest or during treadmill exercise in normal dogs. In dogs with CHF, febuxostat increased LV dP/dtmax at rest and during heavy exercise (P < 0.05), indicating improved LV function with no change of MVO2. Myocardial adenosine triphosphate (ATP) and phosphocreatine (PCr) were examined using 31P nuclear magnetic resonance spectroscopy in the open chest state. In normal dogs, febuxostat increased PCr/ATP during basal conditions and during high workload produced by dobutamine + dopamine (P < 0.05). PCr/ATP was decreased in animals with CHF; in these animals, febuxostat (given after completing basal and high workload measurements with vehicle) tended to increase PCr/ATP during basal conditions with no effect during catecholamine stimulation. Thus, febuxostat improved LV performance in awake dogs with CHF, but caused only a trend toward increased PCr/ATP in the open chest state. It is possible that the antecedent high workload condition prior to drug administration blunted the effect of febuxostat on HEP in the CHF animals. Alternatively, beneficial effects of febuxostat on LV performance in the failing heart may not involve HEP. PMID- 17110809 TI - Resolving the onset of antidepressants' clinical actions: critical for clinical practice and new drug development. PMID- 17110810 TI - Optimizing early prediction for antipsychotic response in schizophrenia. AB - OBJECTIVE: Researchers, by studying first-generation antipsychotics, have established an early prediction model, which had a favorable specificity but a low sensitivity. This study aims to optimize early prediction of treatment response for schizophrenia using a novel statistic method that can be done even under the Microsoft Excel system of a personal computer. METHODS: One hundred twenty-three inpatients with acutely exacerbated schizophrenia were given optimal therapy of risperidone, a commonly used second-generation antipsychotic agent. Response was defined as a reduction of 20% or more in the Positive and Negative Syndrome Scale total score. We applied the generalized estimating equation method's logistic regression to establish an early prediction model based on the treatment results of the first and the second weeks. RESULTS: The proposed method correctly predicted nonresponse at 4 and 6 weeks in 80.8% and 81.8% of the patients, respectively. The method also identified responder at 4 and 6 weeks in 80.0% and 82.8%, respectively. The predictive powers (or correct prediction rates) at 4 and 6 weeks were 80.3% and 82.4%, respectively. In addition, the results based on the responses in Positive and Negative Syndrome Scale scores were slightly better than those in Brief Psychiatric Rating Scale scores. CONCLUSIONS: Using the first 2 weeks' treatment results to predict the fourth or sixth week's treatment response is acceptable in terms of specificity, sensitivity, and predictive power. Further studies are needed. Moreover, whether this model could be applied to establish a prediction system for other psychotropics, such as antidepressants, also deserves research. PMID- 17110811 TI - Predictors of neuroleptic-induced dyskinesia and parkinsonism: the influence of measurement methods and definitions. AB - The accurate and objective measurement of abnormal, involuntary movements remains highly desirable, whether the movements are secondary to pharmacotherapy or an expression of the primary illness. In a previous study, we found that the prevalence of tardive dyskinesia in a sample of 100 subjects ranged from 28% when using the Abnormal Involuntary Movement Scale (AIMS) or the Dyskinesia Identification Scale, Condensed User Version (DISCUS) to 62% using an instrumental measurement (IM) of peripheral dyskinesia. The goal of this study was to examine the relationship between various risk factors for tardive dyskinesia as predictor variables, and the AIMS, DISCUS, and IMs of dyskinesia, tremor, and velocity of motor movement as dependent variables. The sample consisted of 100, mostly patients with schizophrenia. Poor performance on the Mini-Mental State Examination (MMSE) and increasing age were the most consistent predictors of dyskinetic and parkinsonian movements. Various predictors were associated with specific abnormal movements. Head injury was related to slower speed of motor movements and the total DISCUS score. A history of smoking was associated with less IM dyskinesia. For those with coexisting parkinsonism and dyskinesia, significant associations were found with head injury, diabetes mellitus, and an AIMS score of 2 or greater in 2 body areas. Various classes of psychotropic agents seemed to have little influence on the MMSE or the development of dyskinesia and parkinsonism. Increasing age and a lower score on the MMSE seem to be particularly helpful in gauging the risk for parkinsonian and dyskinetic movements. PMID- 17110812 TI - Mortality in elderly dementia patients treated with risperidone. AB - Agitation, aggression, and psychosis are among the most troublesome behavioral and psychological symptoms of dementia (BPSD) and impair the lives of dementia patients and their caregivers. Atypical antipsychotics have been widely prescribed to improve these BPSD. However, in a number of trials with atypical antipsychotics, a consistent increase in overall mortality has been observed. The US Food and Drug Administration issued a warning for all atypical antipsychotics as a result of a meta-analysis of 17 placebo-controlled clinical trials using various atypical antipsychotics for the treatment of BSPD. To evaluate this mortality risk specifically for risperidone, 6 phase-2/3 double-blind trials comparing risperidone with placebo were analyzed. Data were obtained from Johnson & Johnson Pharmaceutical Research and Development. Hazard ratios with 95% confidence intervals were calculated to compare the relative mortality risk between patients treated with risperidone and those treated with placebo. In this meta-analysis, 1721 patients were included. In the pooled sample, the mortality was 4.0% with risperidone versus 3.1% with placebo (relative risk, 1.21; 95% confidence interval, 0.71-2.06) during treatment or within 30 days after treatment discontinuation. The most common adverse events associated with death were pneumonia, cardiac failure or arrest, or cerebrovascular disorder. No relationship was found between risperidone dose and mortality. In conclusion, this meta-analysis found a nonsignificant increase in mortality during treatment with risperidone in dementia patients. Larger studies would be needed to rule out a small increase in mortality in these patients. Careful assessments of potential benefits and risks should be made before prescribing risperidone for the treatment of BPSD. PMID- 17110813 TI - Remission and relapse in the outpatient care of schizophrenia: three-year results from the Schizophrenia Outpatient Health Outcomes study. AB - Remission and relapse are clinical outcomes of increasing interest in schizophrenia. We analyzed remission and relapse, and the sociodemographic and clinical factors associated with these outcomes, in the usual care of schizophrenia using the 3-year, follow-up data from a large cohort of outpatients with schizophrenia taking part in the prospective, observational, European Schizophrenia Outpatient Health Outcomes study. Of the 6516 patients analyzed for remission, 4206 (64.6%) achieved remission during the 3-year, follow-up period. Logistic regression analysis revealed that being female, having a good level of social functioning at study entry, and a shorter duration of illness were factors significantly associated with achieving remission. Treatment with olanzapine was also associated with a higher frequency of remission compared with other antipsychotic agents. A Kaplan-Meier survival curve estimated that relapse occurred in approximately 25% of the patients who achieved remission, with the risk of relapse remaining constant during the follow-up period. Shorter duration of illness, having hostile behaviors, and substance abuse were factors associated with a higher risk of relapse, whereas good level of social functioning and the use of olanzapine and clozapine were associated with a lower risk of relapse. In conclusion, the 3-year results of the Schizophrenia Outpatient Health Outcomes study indicate that the likelihood of remission decreases over the longitudinal course of schizophrenia, but risk of relapse is maintained even after 3 years of achieving remission severity levels. Results suggest that treatment with olanzapine is associated with a better chance of achieving remission than other antipsychotics. Moreover, the use of olanzapine and clozapine is associated with a lower risk of relapse compared with risperidone, quetiapine, and typical antipsychotics. The results should be interpreted conservatively because of the observational, nonrandomized study design. PMID- 17110814 TI - Selegiline transdermal system in the prevention of relapse of major depressive disorder: a 52-week, double-blind, placebo-substitution, parallel-group clinical trial. AB - The selegiline transdermal system (STS) is a monoamine oxidase inhibitor (MAOI) with unique pharmacokinetic and pharmacodynamic properties that was developed to overcome limitations of orally administered MAOIs, particularly dietary tyramine restrictions. We present data from a long-term study assessing the safety and efficacy of initial and continuation STS therapy in patients with major depressive disorder (MDD). After 10 weeks of treatment with STS 6 mg/24 h, 322 patients who responded with a 17-item Hamilton Depression Rating Scale score of 10 or less were randomly assigned to double-blind treatment with STS 6 mg/24 h or placebo for 52 weeks. Relapse was defined as meeting the following criteria on 2 consecutive visits: (1) 17-item Hamilton Depression Rating Scale score of 14 or more, (2) a Clinical Global Impression of Severity score of 3 or more with a 2 point increase from double-blind baseline, and (3) the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for a major depressive episode. At study week 52, significantly fewer STS patients experienced relapse of major depressive episode (25/149 [16.8%]) compared with placebo (50/163 [30.7%]) (P = 0.0025). In addition, patients receiving STS experienced a significantly longer time to relapse compared with those receiving placebo (P = 0.0048). The safety profile of STS was similar to placebo, with the exception of application-site reactions (STS, 15.2%; placebo, 3.7%). No cases of hypertensive crisis were reported, despite the lack of requirement for dietary tyramine restrictions. In conclusion, STS was well tolerated and efficacious in maintaining a sustained response in MDD patients. The results of this study suggest that STS may be suitable in the long-term treatment of MDD. PMID- 17110815 TI - Duloxetine: meta-analyses of suicidal behaviors and ideation in clinical trials for major depressive disorder. AB - BACKGROUND: Uncertainty regarding relationships of antidepressant treatment and suicidality encouraged systematic review of data on suicidal behaviors and ideation from Phase II and III clinical trials of duloxetine for major depressive disorder (MDD). METHODS: We evaluated all completed duloxetine trials in MDD with data lock by February 2, 2004. We compared incidence of suicide-related events with duloxetine versus placebo in controlled trials, using Mantel-Haenszel incidence difference (MHID) and exposure time-adjusted rate difference (MHRD) methods, and analyzed changes in Hamilton Depression Scale (HAMD) Item-3 (suicidality) scores. RESULTS: There were no significant differences in the incidence of suicide-related events with duloxetine versus placebo in 12 placebo controlled trials (duloxetine, 1812; placebo, 1184 [corrected] patients). The MHID for suicide-related behaviors was -0.03% (95% confidence interval [CI], 0.48 to 0.42) and MHRD -0.002 (95% CI, -0.02 to 0.02). Changes in HAMD Item-3 suicidality scores showed more improvement with duloxetine (MHID, 9.56%; 95% CI, 4.50 to 14.6; P < 0.001) and less worsening of suicidal ideation with duloxetine (MHID, -4.25%; 95% CI, -6.55 to -1.95; P < 0.001). Other Item-3 findings showed no consistent pattern; a slightly higher proportion of duloxetine-treated patients with a change from 0 (absent) to 3 was balanced against a higher proportion of placebo-treated patients changing from 0 to 2. CONCLUSIONS: We found no evidence of an increased risk of suicidal behaviors or ideation during treatment with duloxetine compared with placebo in MDD patients. HAMD Item-3 suicidality scores had more improvement and less worsening of suicidal ideation with duloxetine than placebo. PMID- 17110816 TI - Cerebral metabolic effects of intravenous glycine in healthy human subjects. AB - Enhancing N-methyl-D-aspartate (NMDA) receptor function via increasing synaptic concentrations of glycine is currently investigated as a novel approach to treat schizophrenia. The neural correlates of enhanced NMDA receptor function in humans, however, are unclear to date. The present study determines the effects of intravenous administration of the glycine on regional cerebral metabolic rate of glucose (rCMRGlu) in healthy control subjects by using [18F]fluorodeoxyglucose and positron emission tomography and on neuropsychological behavioral measures. Thirteen healthy volunteers were recruited, and 12 subjects completed the protocol. These individuals participated in 1 magnetic resonance imaging study and 2 [18F]fluorodeoxyglucose positron emission tomography studies. In a double blind, randomized, controlled, crossover design, participants received on one test day an intravenous glycine infusion and on the other test day a placebo infusion. There were no significant behavioral and neuropsychological effects of glycine compared with placebo. However, there was a significant reduction of whole-brain CMRGlu during administration of glycine compared with placebo (t = 2.60, df = 11, P = 0.023). In the a priori-selected regions of interest, there was a significant reduction in the cerebellum (t = -3.18, df = 11, P = 0.009) and the dorsolateral prefrontal cortex (t = -2.31, df = 11, P = 0.041). When corrected for whole-brain CMRGlu, rCMRGlu differences were not significant. This study suggests that studies of whole-brain cerebral metabolism may be useful for studying glycine-related mechanisms in healthy humans because there is not a clear cognitive or behavioral signal related to glycine administration at doses thought to be important clinically in patient populations. PMID- 17110817 TI - Efficacy of quetiapine monotherapy in bipolar I and II depression: a double blind, placebo-controlled study (the BOLDER II study). AB - This study evaluated the efficacy and tolerability of quetiapine monotherapy for depressive episodes in patients with bipolar I or II disorder (Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition) who were randomized to 8 weeks of double-blind treatment with quetiapine (300 or 600 mg/d; once daily, evening dosing) or placebo. Patients were assessed weekly using the Montgomery Asberg Depression Rating Scale (MADRS) and Hamilton Depression Rating Scale (HAM D). The primary end point was change in MADRS total score from baseline to Week 8 (analysis of covariance/last-observation-carried-forward analysis). Of 509 patients randomized, 59% completed the study. Improvements from baseline in mean MADRS total scores were significantly greater with quetiapine 300 and 600 mg/d than with placebo from first evaluation (Week 1) through Week 8 (both P 1 per day); (3) drug holidays (>or=1). We found that not only a positive drug attitude, the most important predictor, but also adverse effects, early treatment response, longer onset of depression, and a higher educational level predicted the correct intake of ADs. Extra intakes were predicted by ADs with a shorter half-life and by experience of adverse effects. Drug holidays were predicted by patients with less social support and a negative drug attitude. In conclusion, the predictors of adherence were multifactorial and varied across the 3 adherence types. A positive drug attitude emerged as the most important predictor for correct intakes and drug holidays. PMID- 17110824 TI - Quality of life assessment in adult patients with attention-deficit/hyperactivity disorder treated with atomoxetine. AB - BACKGROUND: Attention-deficit/hyperactivity disorder (ADHD) has its onset during childhood and is estimated to affect 3% to 7% of school-aged children. Unfortunately, the disorder frequently persists into adult life. The burden of this disorder is considerable and is often characterized by academic (or occupational) impairment and dysfunction within the family and society. Despite the existence of research demonstrating the effects of ADHD on certain aspects of life, the clinical trials of treatments for this disorder have focused primarily on efficacy and safety. METHODS: Atomoxetine was approved in the United States in November 2002 for the treatment of ADHD in children, adolescents, and adults. The present study uses data from a clinical trial of atomoxetine in adult patients with ADHD that incorporated a measure of health-related quality of life (the Medical Outcomes Study 36-item short-form health survey [SF-36]) as part of the overall assessment of the success of this relatively new treatment. The primary outcome measure for ADHD symptoms was the Conners Adult ADHD Rating Scale Investigator Rated: Screening Version (CAARS) ADHD total symptom score. RESULTS: In agreement with previous studies, adult patients with ADHD treated with atomoxetine at typical doses showed significant amelioration of ADHD symptoms, as measured on the CAARS. At baseline, the measures of overall mental health (one aspect of quality of life) of adult patients with ADHD were below the average level, as measured on the SF-36. Treatment with atomoxetine significantly improved the measures of mental health and ameliorated the ADHD symptoms. In addition, the 2 measures were correlated. CONCLUSIONS: These data suggest that pharmacological intervention with atomoxetine not only ameliorates ADHD symptoms in adult patients but also improves their perceived quality of life. PMID- 17110825 TI - A randomized, double-blind, placebo-controlled trial of augmentation with an extended release formulation of methylphenidate in outpatients with treatment resistant depression. AB - We examined the efficacy and tolerability of augmentation with an extended release formulation of methylphenidate (OROS MPH, Concerta) in patients with major depression who were nonresponders or partial responders to antidepressants. Sixty subjects with treatment-resistant depression (TRD) participated in a 4 week, randomized, double-blind, placebo-controlled study of augmentation with methylphenidate (18-54 mg/d). The preexisting antidepressant dose was unchanged. The primary efficacy measure was change in the 21-item Hamilton Depression Rating Scale from randomization to end of treatment. Data were analyzed with intent-to treat with last observation carried forward approach. There were no statistically significant differences between the methylphenidate (n = 30) and placebo (n = 30) groups in reduction in 21-item Hamilton Depression Rating Scale scores (drug, 6.9; placebo, -4.7) from baseline to end of treatment (F1,47 = 1.24, P = 0.22), although responders were numerically higher in the extended-release methylphenidate group (40.0%) than in the placebo group (23.3%). On the secondary efficacy measures of changes in Clinical Global Impression-Improvement and Severity scores and Beck Depression Inventory-Second Edition, the drug failed to separate from placebo, although the proportion of responders in the drug group were numerically higher than placebo. There were no significant differences in weight, heart rate, and blood pressure changes between the 2 groups. The common adverse events were loss of appetite, nausea, headache, and anxiety. The mean dose of drug was 34.2 mg/d. The study did not demonstrate a statistically significant benefit for augmentation with methylphenidate in TRD. Combination of methylphenidate with antidepressants was well tolerated. Adequately powered, randomized, controlled trials are necessary to fully evaluate the efficacy of extended-release methylphenidate in TRD. PMID- 17110826 TI - One-year mortality rates of patients receiving methadone and buprenorphine maintenance therapy: a nationally representative cohort study in 2694 patients. AB - Mortality rates in drug-dependent patients in substitution treatment remain a matter of debate. Although several retrospective toxicological or forensic postmortem studies on this issue have been conducted, few prospective studies have addressed this problem. In a nationally representative sample of 2694 opioid dependent patients in substitution treatment either with methadone or buprenorphine at baseline were monitored over a 12-month period (response rate, 91%). A total number of 1629 (60.4%) were still in treatment after 12 months. The overall mortality rate was 1.04%. In total, 28 patients of the initial sample deceased within the 1-year follow-up period. Eleven (0.4%) of these deaths are due to a fatal intoxication. Three patients (0.1%) died of human immunodeficiency virus/acquired immunodeficiency syndrome, and 3 (0.1%) committed suicide. Thirteen of these patients (4 with overdose/polyintoxication) were not in substitution treatment at the time of death. Other reasons included accidents and deaths due to other medical conditions. Only in one case the reason could not be ascertained. The mortality rate was similar in methadone as compared with buprenorphine patients. Taking into account the high comorbidity of opioid dependent patients and the severity of dependence, the mortality rate of approximately 1% confirms that maintenance treatment could be regarded as a fairly safe treatment. PMID- 17110827 TI - n-3 polyunsaturated fatty acids decrease anxiety feelings in a population of substance abusers. AB - There is mounting evidence that low levels of n-3 polyunsaturated fatty acids (PUFAs) play a role in the pathophysiology of a number of psychiatric disorders. Preclinical studies have shown that n-3 PUFAs decrease anxietylike behaviors, but there is a paucity of information about their effects on anxiety in humans. In light of our observation that substance abusers have poor dietary habits and the strong association between anxiety disorders and substance use disorders, the possibility that the administration of supplements of n-3 PUFAs would decrease the anxiety level of a group of substance abusers was explored. Thirteen patients were given on a daily basis capsules containing 3 g of n-3 PUFAS (eicosapentaenoic acid + docosahexaenoic acid). Eleven patients received similarly looking placebo capsules containing vegetable oil. The trial was double blind, randomized, and lasted 3 months. A scale assessing anxiety feelings was administered at baseline and on a monthly basis thereafter. Six PUFA group patients and 8 placebo group patients were followed for an additional 3 months after treatment discontinuation and administered the same questionnaire monthly. Patients who received n-3 PUFAs for 3 months showed a progressive decline in anxiety scores. This was not the case for patients who received placebos. A comparison of the 2 groups was significant (P = 0.010). Anxiety scores remained significantly decreased in the PUFA group for 3 months after treatment discontinuation. A comparison of the 2 groups followed for 6 months was also significant (P = 0.042). In conclusion, these preliminary data indicate that n-3 PUFA supplementation could be beneficial in the treatment of some patients with anxiety disorders. PMID- 17110828 TI - Antipsychotic treatment discontinuation among individuals with schizophrenia and co-occurring substance use. PMID- 17110829 TI - Urinary obstruction with citalopram and aripiprazole combination in an elderly patient. PMID- 17110830 TI - Quetiapine-induced erythema multiforme minor: a case report. PMID- 17110831 TI - Clozapine augmented with aripiprazole in 5 patients with schizophrenia. PMID- 17110832 TI - Appropriateness of plasma level determinations for lithium and valproate in routine care of psychiatric inpatients with affective disorders. PMID- 17110833 TI - Use of a serotonin 1A receptor agonist to treat restless legs syndrome. PMID- 17110834 TI - A case of hyponatremia induced by duloxetine. PMID- 17110835 TI - Possible case of quetiapine-induced rhabdomyolysis in a patient with depression treated with fluoxetine. PMID- 17110836 TI - Delirium associated with concomitant use of low-dose bupropion sustained release and fluoxetine. PMID- 17110837 TI - Epileptiform seizure after sertraline treatment in an adolescent experiencing obsessive-compulsive disorder and presenting a rare pharmacogenetic status. PMID- 17110838 TI - Serotonin syndrome precipitated by linezolid in a medically ill patient on duloxetine. PMID- 17110839 TI - Alcohol abuse in an elderly woman taking donepezil for Alzheimer disease. PMID- 17110840 TI - Beneficial effects of the glutamate-modulating agent riluzole on disordered eating and pathological skin-picking behaviors. PMID- 17110841 TI - Methadone and the QT interval: relations to the serum concentrations of methadone and its enantiomers (R)-methadone and (S)-methadone. PMID- 17110842 TI - Inhaled cocaine used to relieve "off" periods in patients with Parkinson disease and unpredictable motor fluctuations: a report of 2 cases. PMID- 17110844 TI - Comments on guest editorial, "Chronic fatigue syndrome, mast cells, and tricyclic antidepressants". PMID- 17110847 TI - Ills we know not of. PMID- 17110848 TI - Inhalational anthrax. AB - Inhalational anthrax is a lethal infection acquired from the inhalation of Bacillus anthracis, a pathogen classified as a Category A bioterrorist agent by the Centers for Disease Control and Prevention. The recent 2001 attack in which weaponized spores were delivered by mail to several US cities exposed our vulnerability to bioterrorism, and taught us important lessons in the timely diagnosis of this devastating disease. It is clear that patient mortality is significantly decreased by early recognition and immediate administration of antibiotic therapy. Unfortunately, the nonspecific clinical presentation is often misinterpreted as a flu-like illness and confirmatory microbiologic tests may take up to 24 hours. Radiologic manifestations, however, are distinctive and may prove essential in directing appropriate clinical care in the critical early hours of inhalational anthrax. PMID- 17110849 TI - The radiologic manifestations of H5N1 avian influenza. AB - Avian influenza is caused by the H5N1 subtype of the influenza A virus. Human transmission is either directly through close contact with infected birds usually poultry or their secretions. To date 178 people throughout South East Asia have been infected with 85 deaths. Patients usually present with a rapidly progressive pneumonia that can result in respiratory failure and acute respiratory distress syndrome. The chest radiograph therefore remains the most convenient and accessible imaging modality. Studies have shown that most radiographs are abnormal at the time of presentation with multifocal consolidation the commonest radiographic finding. During the course of disease, pleural effusions and cavitation can also develop. Consolidation that involves > or = 4 zones on presentation or at day 7 after the onset of symptoms and subsequent development of acute respiratory distress syndrome are generally associated with an adverse outcome. Chest CT examinations performed during the convalescent period have demonstrated persistent ground glass attenuation and segmental consolidation. Additional features included pseudocavitation, pneumatocoele formation, lymphadenopathy, and centrilobular nodules. Overall the appearances are suggestive of mild fibrosis. PMID- 17110850 TI - Thoracic radiology of infections emerging after natural disasters. AB - When natural disasters demolish shelter, destroy sources of clean drinking water, and disrupt the availability of medical care, vast numbers of people are placed at increased risk of disease. The infectious diseases that propagate under these conditions are usually common ones. Occasionally, a natural disaster alters the local environment in ways that markedly increase the prevalence of a disease that is endemic to a geographic region, occurring only as isolated cases under normal conditions. Many of these infections may affect the thorax. In this article, we discuss the radiologic findings of 4 infectious diseases, coccidioidomycosis, leptospirosis, melioidosis, and Chagas disease, which may flourish after natural disasters strike areas where they are endemic. PMID- 17110851 TI - Radiology of severe acute respiratory syndrome (SARS): the emerging pathologic radiologic correlates of an emerging disease. AB - Severe acute respiratory distress syndrome (SARS) caused by SARS-associated coronavirus (SARS-CoV) is a systemic infection that clinically manifests as progressive pneumonia. During the initial phases of infection the virus causes pauci-inflammatory alveolar and interstitial edema that result in imaging abnormalities dominated by ground glass opacities (GGO). Severe SARS cases can develop radiologic and pathologic findings of diffuse alveolar damage. Although radiologic evidence of acute bronchiolitis is absent, SARS-CoV also infects ciliated airway epithelium, probably accounting for respiratory transmissibility of the virus. Radiologic recovery from SARS can be complete, but computed tomography images often show persistent GGO and reticular opacities, some of which reflect pathologic findings of fibrosis. Long-term follow-up imaging of survivors shows gradual decrease of GGO and reticulation with persistent air trapping in some patients. The latter is evidence of small airway disease that is not radiologically evident at the onset of the disease. PMID- 17110852 TI - Counting ribs on CT by assessing costal attachments to the proximal xiphoid: is this method accurate? AB - OBJECTIVES: To evaluate the usefulness of the method of counting ribs by assessing anatomic variations of the attachments of costal cartilages to the proximal xiphoid. MATERIALS AND METHODS: From January to September 2005, 224 subjects (136 men, 88 women, age 13 to 89 years, mean age 55 years) underwent computed tomography examination of the chest. Axial images of the chest were obtained on a 16-slice multidetector computed tomography. Counting ribs was performed by using the medial clavicle as an anatomic landmark to identify the first costal cartilage. We analyzed variety and incidence of the attachment patterns of costal cartilages to the proximal xiphoid. RESULT: Out of the 224 patients, the last costal attachments to the proximal xiphoid were the sixth costal cartilages bilaterally for 2 (0.9%) subjects, one 6th and one 7th for 4 (1.8%) subjects, bilateral seventh for 191 (85.3%) subjects, one 7th and one 8th for 15 (6.7%) subjects, and bilateral eighth for 12 (5.4%) subjects. CONCLUSIONS: The method of counting ribs from the proximal xiphoid is inaccurate because the sixth, seventh, and eighth costal cartilages may each attach to the proximal xiphoid. PMID- 17110853 TI - Thoracic lymph node enlargement in usual interstitial pneumonitis and nonspecific interstitial pneumonitis: prevalence, correlation with disease activity and temporal evolution. AB - PURPOSE: To evaluate the prevalence of thoracic lymph node enlargement (LNE) in usual (UIP) and nonspecific (NSIP) interstitial pneumonitis, change in LNE over time, and if LNE is related to disease activity. METHODS AND MATERIALS: High resolution CT scans (HRCT) in 20 patients each with UIP and NSIP were retrospectively reviewed. Two HRCT scans were reviewed for each patient, at diagnosis and a mean of 1 +/- 0.7 years later. Two thoracic radiologists independently recorded the location and size of thoracic lymph nodes (LNs) > 10 mm in short-axis diameter, using the American Thoracic Society lymph node mapping scheme. HRCT disease severity was scored for ground glass opacity and fibrosis. The number and size of enlarged LN stations were compared with HRCT scores. RESULTS: LNE was found on 44 HRCT examinations (21 baseline prevalence 52.5%, 23 follow-up, prevalence 57.5%), most common in the low right paratracheal (38%) and subcarinal (36%) regions. There was no significant difference in LN size or number of enlarged LN stations between baseline and follow-up CT. LNE prevalence was not different on baseline CT (P = 0.34) follow-up CT (P = 0.11) between UIP and NSIP patients. The mean size of the largest enlarged LN was 1.36 cm (1 to 2.1 cm) at baseline and 1.43 cm (1 to 1.9 cm) on follow-up CT. Mean CT ground glass and fibrosis scores were 1.98 and 1.6 when LNE was present, and 1.34 and 1.03 when absent (P = 0.008 and 0.003, respectively). The number and maximum size of enlarged LNs did not correlate with CT ground glass or fibrosis scores. Five patients who developed LNE between baseline and follow-up CT examinations had a greater increase in CT fibrosis scores than patients whose LNE status did not change (P = 0.004); CT ground glass scores were not significantly different. There was a trend for UIP patients to progress from absence of LNE to presence of LNE (4/20 patients or 20%). CONCLUSIONS: Intrathoracic LNE is common in both UIP and NSIP, and becomes increasingly prevalent in UIP patients over time. LNE is more prevalent with more severe lung disease. An increase in LNE over time is associated with the progression of fibrosis, and should not raise concern for co existing infection or malignancy, in the absence of other clinical findings that would suggest this. PMID- 17110854 TI - An adult case of bilateral true tracheal bronchi associated with hemoptysis. AB - The term tracheal bronchus encompasses a variety of bronchial anomalies originating from the trachea or main bronchus and directed to the upper lobe, and a true tracheal bronchus is any bronchus originating from the trachea. In most cases, it is found incidentally during bronchoscopy or tomography. It is often unilateral, and bilateral true tracheal bronchi are very rare. We report a case of a 54-year-old woman who had bilateral true tracheal bronchi associated with hemoptysis. PMID- 17110855 TI - Hypoplasia of the descending thoracic and abdominal aorta: a case report and review of literature. AB - Coarctation of the aorta has been found at necropsy in approximately 1 of every 1550 individuals. The most common location for segmental coarctation (juxtaductal) is the attachment of the ductus arteriosus to the thoracic aorta, which accounts for 98% of focal lesions. Less commonly, a relatively long segment of constriction extends beyond the left subclavian artery. When longer segments of the aorta are narrowed, the term "hypoplasia" is often used. Aortic hypoplasia, an exceedingly rare cardiovascular anomaly, has been described in all portions of the thoracic and abdominal aorta. In the current case report, we described a 21-year-old man presenting with severe hypertension in whom the diagnosis of hypoplasia of the descending thoracic and abdominal aorta was made using with magnetic resonance imaging. PMID- 17110856 TI - Multidetector-row computed tomography of a Valsalva sinus aneurysm in a patient with Behcet disease. AB - Behcet disease is a multisystem disease, which can cause cardiovascular manifestations such as arterial occlusion or aneurysm. This report describes multidetector-row computed tomography findings of an unruptured aneurysm of the left Valsalva sinus which was associated with Behcet disease. A 45-year-old woman with Behcet disease had an aneurysm of the left Valsalva sinus which constricted the left anterior descending artery. The multidetector-row computed tomography was useful in detecting the aneurysm and complications, and was beneficial in the evaluation of patency in the coronary arteries after surgery. PMID- 17110857 TI - Dual energy subtraction digital radiography: technical considerations, clinical applications, and imaging pitfalls. AB - Emerging applications in digital radiography offer exciting advances in the evaluation of lung disease. Dual energy subtraction techniques have been recognized as a promising technique in the detection and characterization of pulmonary nodules. This essay will demonstrate a significantly expanded diagnostic role in the assessment of cardiothoracic disease with dual energy subtraction and digital radiography. PMID- 17110858 TI - Impact of domestic violence posters on female caregivers' opinions about domestic violence screening and disclosure in a pediatric emergency department. AB - OBJECTIVES: The objectives of this study were to examine female caregivers' attitudes about the display of domestic violence (DV) resources in a pediatric emergency department (ED) and to explore whether these resources engendered positive feelings about DV screening and encouraged disclosure. METHODS: We conducted a cross-sectional survey in a pediatric ED during 2 distinct periods, comparing responses of female caregivers before (pre) and after (post) displaying DV posters and cards. Women were surveyed about (1) personal experience with DV, (2) the appropriateness of DV posters and screening in a pediatric ED, and (3) willingness to divulge DV, if abused. RESULTS: The 2 groups (pre, n = 133; post, n = 136) did not significantly differ with respect to age, race, education, or personal DV history. The majority endorsed that "it is appropriate to have DV posters," with the post group responding in this manner more often than the pre group (pre, 85%; post, 95%; odds ratio [OR], 3.3; 95% confidence interval [CI], 1.3-8.5). The post group was less likely to prefer pediatric ED DV screening (pre, 76%; post, 63%; OR, 0.5; 95% CI, 0.3-0.9) and tended to be less likely to say that they would divulge (pre, 85%; post, 75%; OR, 0.6; 95% CI, 0.3-1.1). In both groups, women with a DV history were less likely than women without this history to say that they would disclose DV to their pediatric ED provider (P < 0.001). CONCLUSIONS: These results suggest the need for further exploration of how to most effectively help and provide resources for abused women in this setting. PMID- 17110859 TI - Preparedness of selected pediatric offices to respond to critical emergencies in children. AB - OBJECTIVE: To determine the preparedness of pediatric offices that had activated emergency medical services (EMS) for a critically ill child requiring airway management. METHODS: Fifteen patients who initially presented to pediatric or family practice offices but required EMS activation and cardiac and/or respiratory support were identified from a previous prospective study of airway management in children. Two to 4 years after the emergency requiring EMS activation, the offices were contacted to complete a written survey about office preparedness for pediatric emergencies. RESULTS: Eight of 15 offices (53%) returned a survey. Pediatricians staffed all responding offices, and all offices were within 5 miles of an emergency department. Airway emergencies were the most common emergencies seen in the offices. Availability of emergency equipment and medications varied. All offices stocked albuterol, and most (7/8) had an oxygen source with a flowmeter. However, only half of the offices had a fast-acting anticonvulsant, and a quarter had no anticonvulsant. Three offices lacked bag mask (manual) resuscitators with all appropriate sized masks, and 3 offices lacked suction. The most common reasons cited for not stocking all emergency equipment and drugs were quick response time of EMS and proximity to an emergency department. CONCLUSIONS: Even after treating a critically ill child who required advanced cardiac and/or pulmonary support, offices were ill prepared to handle another serious pediatric illness or injury. PMID- 17110860 TI - Survey of parental willingness to pay and willingness to stay for "painless" intravenous catheter placement. AB - OBJECTIVES: This study examined how much parents are willing to pay and/or willing to stay to make their child's intravenous (IV) catheter placement painless. METHODS: A prospective survey was conducted using a questionnaire administered to a consecutive sample of parents presenting to an emergency department (ED). Eligible subjects were parents accompanying a child 8 years of age or younger. A hypothetical visit to the ED, requiring an IV for their child, was described. Parents were asked if they would prefer to make the IV catheter placement painless and if so, how much of an increase in out-of-pocket cost (none, 15 dollars, and 100 dollars) and/or length of stay they would be willing to incur (no time, 15 minutes, 1 hour). Statistics were chiefly descriptive. Associations of demographic elements with willingness to pay and willingness to stay were analyzed using chi and t tests, where appropriate. RESULTS: One hundred eight subjects were available for analysis. Most parents were mothers (71%), white (53%), and with previous IVs (70%). Most children were boys (55%) with no previous IV placements (55%). The choice of a painless IV placement was independent of demographics and IV experience. Most parents (89%) chose a painless IV placement. Of these parents, 65% chose a willingness to stay of 1 extra hour, and 77% a willingness to pay at least 15 dollars; 37% of parents would pay 100 dollars. Willingness to pay was dependent on both income (P = 0.014) and ethnicity (P = 0.0013). Willingness to stay was independent of both income (P = 0.24) and ethnicity (P = 0.07). CONCLUSIONS: Parents are willing to spend both time and money to make their child's IV placement painless. This information should be considered when choosing therapies to reduce the pain of IV placement. PMID- 17110861 TI - Children in taxis: an opportunity for pediatricians and emergency physicians to save lives? AB - OBJECTIVE: Child safety devices (infant seats, booster seats, and seat belts) are effective in curbing the risk of injury; however, there remains a pattern of parental nonuse or misuse of safety seats. The aim of this study was to assess the level of knowledge and compliance of parents with children presenting for emergency care of the National Highway and Traffic Association safety seat guidelines in private cars and taxicabs. METHODS: Two hundred forty-two caregivers of children (ages range, 2 weeks to 19 years) presenting for care in the pediatric emergency department of an urban university hospital were approached to complete an interviewer-administered questionnaire, and 225 participated. The questionnaire included knowledge, attitude, and behavior questions on protective equipment for various aged children. RESULTS: Eleven (47.8%) of 23 children 1 year or younger were reported to use infant seats often or always while riding in private cars, compared with 8 (22.2%) of 36 children 1 year or younger were reported to their use while in taxis (P < 0.05). Seventeen (85%) of 20 children older than 8 years were reported to have used seat belts often or always in private cars versus 10 (41.7%) of 24 in taxis (P < 0.01). One hundred fifty-four (99.3%) of 155 subjects knew the National Highway Traffic Safety Administration recommended position for the safety seat for their child. Most parents believed in the efficacy of child safety seats in preventing vehicle injuries and reported they would be more likely to use safety devices if they received information on their use in the emergency department. CONCLUSIONS: Data from this survey show that use of safety seats is lower in taxis than in private automobiles and that this is attributable to the inconvenience of carrying these seats to and from the taxi rather than financial considerations or lack of knowledge about their effectiveness. Strategies should be sought to increase availability of child safety devices in taxicabs. The emergency department, as well as the pediatrician's office or clinic, can be a locus for an educational intervention to parents and caregivers on child passenger safety. PMID- 17110862 TI - Endotracheal tube size estimation for children with pathological short stature. AB - OBJECTIVE: To compare length-based estimates of endotracheal tube (ETT) size and age-based estimates with anesthesiologist-selected ideal ETT size in children with medical conditions affecting normal growth, known as pathological short stature (PSS). METHODS: We conducted a retrospective review of the anesthesia database of all children undergoing tracheal intubation for any surgical procedure during a 3-year period. The anesthesiologist-selected ideal ETT size was defined as that selected and successfully used throughout the case under the supervision of a board-certified pediatric anesthesiologist. Objective criteria, such as leak test and adequate oxygenation/ventilation, were used to validate the appropriateness of the ETT chosen. For analysis, the children were classified as normal length for age versus PSS, defined as less than 5% length for age on the Centers for Disease Control and Prevention growth chart. The proportions of clinically relevant predicted ETTs, within +/-0.5 mm of the anesthesiologist selected ideal ETT size, based on both age- and length-based formulas for each group were then compared. RESULTS: Five thousand one hundred seventy-five patient records were analyzed. In children with normal stature, age-predicted ETT size was within the clinically relevant range in 89.8% (95% confidence interval [CI], 88.9%-90.7%), and length-predicted ETT size was within the clinically relevant range in 92.8% (95% CI, 92.0%-93.6%). In children with PSS, age-predicted ETT size was within the clinically relevant range in 86.6% (95% CI, 84.3%-89.0%), and length-predicted ETT size was within the clinically relevant range in 92.2% (95% CI, 90.3%-94.0%). The correlation coefficient for age to anesthesiologist selected ideal ETT size was strong for both normal and PSS patients (r = 0.91 and r = 0.93, respectively). Length was also highly correlated to actual ETT size used for both groups (r = .91). CONCLUSIONS: Length-based prediction of ETT size is at least as accurate as age-based estimation in both normal and pathologically short children. PMID- 17110863 TI - A febrile child with seizure and hemiparesis. AB - Febrile seizures are the most common neurological disorders in children and are among the more common symptoms that lead to an emergency department visit. Although most febrile seizures are simple and benign, these seizures can infrequently create a diagnostic dilemma. The diagnosis of cerebral venous thrombosis is challenging to emergency physicians because it can mimic the presentation of many other disorders, including ischemic and hemorrhagic stroke, tumor, and abscess. In addition, the broad variety of signs and symptoms makes the clinical diagnosis difficult. The patients may be presented with signs of increased intracranial pressure or focal neurological deficits. It is an uncommon but potentially dangerous cause of hemiparesis after seizure. Early recognition of this condition and appropriate management may reduce the mortality rate. We present a young child with dural sinus thrombosis who presented with seizures associated with fever and subsequent hemiparesis, and explained a possible mechanism of focal neurological deficit. PMID- 17110864 TI - Hypocalciuric hypercalcemia presenting as neonatal rib fractures: a newly described mutation of the calcium-sensing receptor gene. AB - A 2-week-old infant presented with bilateral rib fractures, hypercalcemia, and subperiosteal bone erosions. Parathyroid hormone levels were elevated and urine calcium low. Her parent's laboratory test results were normal. Gene sequencing revealed a new mutation of the calcium-sensing receptor gene, causing severe neonatal hyperparathyroidism, a variant of hypocalciuric hypercalcemia. This is a rare cause of neonatal hyperparathyroidism and nonabusive fractures. PMID- 17110865 TI - Hyponatremic dehydration as a presentation of cystic fibrosis. AB - BACKGROUND: The purpose of this study is to present a case report of a child with hyponatremic dehydration diagnosed after CF and to review the cases of 13 patients with CF who had the same initial presentation in our hospital. METHODS: This report reviewed the clinical records of children diagnosed with CF to ascertain the prevalence of metabolic alkalosis with electrolyte depletion as the presentation of CF. It also used sweat tests to diagnose a child with CF. RESULTS: The laboratory tests of a 12-month-old girl presented 3 times to the ;pediatric emergency department with vomiting and weight loss showed hyponatremia, hypochloremia, and metabolic alkalosis. The patient was subsequently diagnosed with CF by means of 2 positive sweat tests. Meanwhile, the review of the clinical records of all children diagnosed with CF from 1985 to 2004 (N = 77) showed that the prevalence of metabolic alkalosis with electrolyte depletion as the presentation of CF was 16.8%. The age of the infants ranged from 3 to 14 months. All episodes took place during summer. CONCLUSIONS: There are not many causes of metabolic alkalosis with hyponatremic dehydration, and one of them is CF. This report emphasizes sodium depletion as a common sign of CF presentation. This is most important in countries where the neonatal screening test for CF is not available because the disease may be asymptomatic or oligosymptomatic for several months or even years. Cystic fibrosis should be considered in differential diagnosis of any child presenting with unexplained hyponatremic dehydration. PMID- 17110866 TI - An infant with tachypnea. AB - The purpose of this case report is to illustrate the diagnostic difficulties of congestive heart failure in an infant. When presenting to the emergency department, these patients are often evaluated for sepsis, congenital heart disease, metabolic disorders, and myocarditis. We report a case of a 3(1/2)-month old male who presented to the pediatric emergency department with congestive heart failure. He was found to have vitamin D deficiency rickets induced cardiomyopathy. PMID- 17110867 TI - ECGs in the ED. PMID- 17110868 TI - Vomiting and diarrhea in an infant: a startling abdominal radiograph. PMID- 17110869 TI - Pediatric emergency medicine: legal briefs. PMID- 17110870 TI - Update on antidotes for pediatric poisoning. AB - Antidotes are playing an increasing role in therapy for pediatric poisonings. Although initial response to all pediatric poisonings begins with basic stabilization, knowledge of specific antidotes, their mechanisms of action, safety profile in pediatrics, and dosing regimens can be life-saving for pediatric victims of nerve gas exposure, acetaminophen toxicity, methanol and ethylene glycol ingestion, and snakebites. This article presents an overview of the pathophysiology, symptoms, antidotes, and emergency management of these toxicological emergencies. PMID- 17110872 TI - Use of dynamic compliance for open lung positive end-expiratory pressure titration in an experimental study. AB - OBJECTIVE: We tested whether the continuous monitoring of dynamic compliance could become a useful bedside tool for detecting the beginning of collapse of a fully recruited lung. DESIGN: Prospective laboratory animal investigation. SETTING: Clinical physiology research laboratory, University of Uppsala, Sweden. SUBJECTS: Eight pigs submitted to repeated lung lavages. INTERVENTIONS: Lung recruitment maneuver, the effect of which was confirmed by predefined oxygenation, lung mechanics, and computed tomography scan criteria, was followed by a positive end-expiratory pressure (PEEP) reduction trial in a volume control mode with a tidal volume of 6 mL/kg. Every 10 mins, PEEP was reduced in steps of 2 cm H2O starting from 24 cm H2O. During PEEP reduction, lung collapse was defined by the maximum dynamic compliance value after which a first measurable decrease occurred. Open lung PEEP according to dynamic compliance was then defined as the level of PEEP before the point of collapse. This value was compared with oxygenation (Pao2) and CT scans. MEASUREMENTS AND MAIN RESULTS: Pao2 and dynamic compliance were monitored continuously, whereas computed tomography scans were obtained at the end of each pressure step. Collapse defined by dynamic compliance occurred at a PEEP of 14 cm H2O. This level coincided with the oxygenation-based collapse point when also shunt started to increase and occurred one step before the percentage of nonaerated tissue on the computed tomography exceeded 5%. Open lung PEEP was thus at 16 cm H2O, the level at which oxygenation and computed tomography scan confirmed a fully open, not yet collapsed lung condition. CONCLUSIONS: In this experimental model, the continuous monitoring of dynamic compliance identified the beginning of collapse after lung recruitment. These findings were confirmed by oxygenation and computed tomography scans. This method might become a valuable bedside tool for identifying the level of PEEP that prevents end-expiratory collapse. PMID- 17110873 TI - Angiopoietin-2 is increased in severe sepsis: correlation with inflammatory mediators. AB - OBJECTIVE: Angiopoietin (Ang)-2 is an endothelium-specific growth factor, regulated by proinflammatory stimuli, that destabilizes vascular endothelium and increases vascular leakage; consequently, Ang-2 may contribute to sepsis pathophysiology. We have studied 1) serum Ang-2 levels in critically-ill patients and investigated potential relationships with inflammatory mediators and indices of disease severity and 2) the effect of sepsis-related inflammatory mediators on Ang-2 production by lung endothelium in vitro. DESIGN: Prospective clinical study followed by cell culture studies. SETTING: General intensive care unit and research laboratory of a university hospital. SUBJECTS: Human and bovine lung microvascular endothelial cells and 61 patients (32 men). Patients were grouped according to their septic stage as having: no systemic inflammatory response syndrome (n = 6), systemic inflammatory response syndrome (n = 8), sepsis (n = 16), severe sepsis (n = 18), and septic shock (n = 13). INTERVENTIONS: Cells were exposed to lipopolysaccharide, tumor necrosis factor-alpha, and interleukin-6. MEASUREMENTS AND MAIN RESULTS: Patients' serum Ang-2 levels were significantly increased in severe sepsis as compared with patients with no systemic inflammatory response syndrome or sepsis (p < .05 by analysis of variance). Positive linear relationships were observed with: serum tumor necrosis factor alpha (rs = 0.654, p < .001), serum interleukin-6 (rs = 0.464, p < .001), Acute Physiology and Chronic Health Evaluation II score (rs = 0.387, p < .001), and Sequential Organ Failure Assessment score (rs = 0.428, p < .001). Multiple regression analysis revealed that serum Ang-2 is mostly related to serum tumor necrosis factor-alpha and severe sepsis. Treatment of human lung microvascular endothelial cells with all mediators resulted in a concentration-dependent Ang-2 reduction. Treatment of bovine lung microvascular endothelial cells with lipopolysaccharide and tumor necrosis factor-alpha increased Ang-2 release, and interleukin-6 reduced basal Ang-2 levels. CONCLUSIONS: First, patients' serum Ang 2 levels are increased during severe sepsis and associated with disease severity. The strong relationship of serum Ang-2 with serum tumor necrosis factor-alpha suggests that the latter may participate in the regulation of Ang-2 production in sepsis. Second, inflammatory mediators reduce Ang-2 release from human lung microvascular endothelial cells, implying that this vascular bed may not be the source of increased Ang-2 in human sepsis. PMID- 17110874 TI - Mortality in Emergency Department Sepsis (MEDS) score predicts 1-year mortality. AB - OBJECTIVE: To assess the predictive performance for 1-yr mortality of the previously derived and validated Mortality in Emergency Department Sepsis (MEDS) score. DESIGN: Prospective cohort study. PATIENTS: Consecutive adult (aged > or =18 yrs) emergency department patients presenting to an urban, tertiary care, university hospital were eligible if they had a clinically suspected infection as indicated by the decision to obtain a blood culture. The enrollment period was between February 1, 2000, and February 1, 2001. Of 3,926 eligible patient visits, 3,762 (96%) were enrolled and 3,102 unique first visits were analyzed. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A total of 667 patients (21.5%) died within 1 yr. The unadjusted 1-yr mortality rates for the MEDS risk groups were: very low risk, 7%; low risk, 20%; moderate risk, 37%; high risk, 64%; very high risk, 80%. Using a Cox proportional hazard model that controlled for age, sex, and Charlson co-morbidity index, the 1-yr hazard ratios compared with the baseline very low-risk group were: low risk, 2.2 (1.7-2.9); moderate risk, 3.5 (2.7-4.6); high risk, 6.7 (4.9-9.3); and very high risk, 10.5 (7.2 15.4). The groups were significantly different (p < .0001). CONCLUSIONS: Although the score was initially derived for 28-day in-hospital mortality, our results indicate that the MEDS score also predicts patient survival at 1 yr after index hospital visit with suspected infection. The score needs external validation before widespread use. PMID- 17110875 TI - Negative mesenteric effects of lung recruitment maneuvers in oleic acid lung injury are transient and short lasting. AB - OBJECTIVE: To test the hypothesis that repeated recruitment maneuvers (RMs) have sustained negative effects on mesenteric circulation, metabolism, and oxygenation 60 mins after RMs in pigs with oleic acid lung injury. Further, we aimed to test the hypothesis that an infusion of prostacyclin (PC) at 33 ng.kg.min would attenuate such possible negative mesenteric effects. DESIGN: Randomized, experimental, controlled study. SETTING: University hospital animal laboratory. SUBJECTS: A total of 31 anesthetized, fluid-resuscitated pigs with oleic acid lung injury. INTERVENTIONS: : Animals were randomized to one of the following four groups: a control group (n = 7) that received no intervention, recruitment group (n = 8) that underwent the RM sequence, a prostacyclin group (n = 8) that received an infusion of PC, and a recruitment-prostacyclin group (n = 8) that received an infusion of PC and concomitant RM sequence. MEASUREMENTS AND MAIN RESULTS: We measured systemic and mesenteric hemodynamic variables, jejunal mucosal perfusion, mesenteric lactate flux, jejunal tissue oxygen tension, and mesenteric oxygen delivery, uptake, and extraction ratio. Five minutes after RMs, mesenteric oxygen extraction ratio and mesenteric lactate flux were more prominently increased in the recruitment group, giving evidence of worsened mesenteric conditions after RMs. These signs of worsened conditions were further supported by more decreased jejunal tissue oxygen tension and portal vein oxygen saturation in the recruitment group. PC preserved mesenteric oxygenation, as indicated by less of a decrease in portal vein oxygen saturation at the time corresponding to 5 mins after RM and less of a decrease in mesenteric oxygen delivery at the time corresponding to 15 mins after RM. PC preserved mesenteric oxygenation as indicated by less of a decrease in portal vein oxygen saturation at 5 mins after RM and an attenuated increase in mesenteric oxygen extraction ratio at 5 mins after RM. There was a trend toward worsened jejunal mucosal perfusion, although not significant. CONCLUSIONS: In an oleic acid lung injury model, three repeated RMs did not improve systemic oxygenation or lung mechanics. Negative effects on mesenteric oxygenation and metabolism were transient and short lasting. The intestinal effects of PC during RMs were minor and opposing, showing preserved oxygenation but a trend toward worsened mucosal perfusion. PMID- 17110876 TI - Perceptions of safety culture vary across the intensive care units of a single institution. AB - OBJECTIVE: To determine whether safety culture factors varied across the intensive care units (ICUs) of a single hospital, between nurses and physicians, and to explore ICU nursing directors' perceptions of their personnel's attitudes. DESIGN: Cross-sectional surveys using the Safety Attitudes Questionnaire-ICU version, a validated, aviation industry-based safety culture survey instrument. It assesses culture across six factors: teamwork climate, perceptions of management, safety climate, stress recognition, job satisfaction, and work environment. SETTING: Four ICUs in one tertiary care hospital. SUBJECTS: All ICU personnel. MEASUREMENTS AND MAIN RESULTS: We conducted the survey from January 1 to April 1, 2003, and achieved a 70.2% response rate (318 of 453). We calculated safety culture factor mean and percent-positive scores (percentage of respondents with a mean score of > or =75 on a 0-100 scale for which 100 is best) for each ICU. We compared mean ICU scores by ANOVA and percent-positive scores by chi square. Mean and percent-positive scores by job category were modeled using a generalized estimating equations approach and compared using Wald statistics. We asked ICU nursing directors to estimate their personnel's mean scores and generated ratios of their estimates to the actual scores.Overall, factor scores were low to moderate across all factors (range across ICUs: 43.4-74.9 mean scores, 8.6-69.4 percent positive). Mean and percent-positive scores differed significantly (p < .0083, Bonferroni correction) across ICUs, except for stress recognition, which was uniformly low. Compared with physicians, nurses had significantly lower mean working conditions and perceptions of management scores. ICU nursing directors tended to overestimate their personnel's attitudes. This was greatest for teamwork, for which all director estimates exceeded actual scores, with a mean overestimate of 16%. CONCLUSIONS: Significant safety culture variation exists across ICUs of a single hospital. ICU nursing directors tend to overestimate their personnel's attitudes, particularly for teamwork. Culture assessments based on institutional level analysis or director opinion may be flawed. PMID- 17110877 TI - Cost of Gram-negative resistance. AB - OBJECTIVE: It is unclear that infections with Gram-negative rods resistant to at least one major class of antibiotics (rGNR) have a greater effect on patient morbidity than infections caused by sensitive strains (sGNR). We wished to test the hypothesis that rGNR infections are associated with higher resource utilization. DESIGN: Retrospective observational cohort study of prospectively collected data. SETTING: University hospital surgical intensive care unit and ward. PATIENTS: Surgical patients with at least one GNR infection. MEASUREMENTS: We compared admissions treated for rGNR infection with those with sGNR infections. Primary outcomes were total hospital costs and hospital length of stay. Other outcomes included antibiotic treatment cost, in-hospital death, and intensive care unit length of stay. After univariate analysis comparing outcomes after rGNR infection with those after sGNR infection, multivariate linear regression models for hospital cost and length of stay were created to account for potential confounders. MAIN RESULTS: Cost data were available for 604 surgical admissions treated for at least one GNR infection (Centers for Disease Control and Prevention criteria), 137 (23%) of which were rGNR infections. Admissions with rGNR infections were associated with a higher severity of illness at the time of infection (Acute Physiology and Chronic Health Evaluation II score, 17.6 +/- 0.6 vs. 13.9 +/- 0.3), had higher median hospital costs ($80,500 vs. $29,604, p < .0001) and median antibiotic costs ($2,607 vs. $758, p < .0001), and had longer median hospital length of stay (29 vs. 13 days, p < .0001) and median intensive care unit length of stay (13 days vs. 1 day, p < .0001). Infection with rGNR within the first 7 days of admission was independently predictive of increased hospital cost (incremental increase in median hospital cost estimated at $11,075; 95% confidence interval, $3,282-$20,099). CONCLUSIONS: Early infection with rGNR is associated with a high economic burden, which is in part related to increased antibiotic utilization compared with infection with sensitive organisms. Efforts to control overuse of antibiotics should be pursued. PMID- 17110878 TI - Surgical repair of tricuspid valve regurgitation caused by blunt thoracic trauma. PMID- 17110879 TI - Endovascular management of trauma related renal artery thrombosis. PMID- 17110880 TI - Sonographic depiction of the needle decompression of a tension hemo/pneumothorax. PMID- 17110881 TI - Second course of recombinant human activated protein C delivered to a severely septic patient after recent surgery. PMID- 17110882 TI - High-voltage electrical brain injury. PMID- 17110883 TI - Video-assisted thoracoscopic retrieval of an intrapleural foreign body after penetrating chest trauma. PMID- 17110884 TI - Acquired traumatic imperforate anus-management of a complex rectovesical fistula. PMID- 17110885 TI - Suffocation caused by a foreign body in the upper intra-thoracic esophagus. PMID- 17110886 TI - Acute traumatic spondylolisthesis at the lumbosacral junction. PMID- 17110887 TI - Intrathoracic fracture-dislocation of the proximal humerus: a case report and report of a new surgical technique. PMID- 17110888 TI - Complete aortic rupture in a polytrauma patient: damage control orthopaedics. PMID- 17110889 TI - Using expanded polytetrafluoroethylene patches to repair perforations of the gastrointestinal tract: a two-case report. PMID- 17110890 TI - Type v physeal injury. PMID- 17110891 TI - Acute posterior fossa isodense epidural hematoma: diagnostic pitfalls on computed tomography. PMID- 17110892 TI - Successful topical therapy with voriconazole: pseudallescheriasis after injury. PMID- 17110893 TI - Life-threatening bleeding of bilateral maxillary arteries in maxillofacial trauma: report of two cases. PMID- 17110894 TI - Adipose tissue-derived cells: from physiology to regenerative medicine. AB - During the last past years, the importance and the role of adipose tissues have been greatly expanded. After finding that adipose tissues are metabolically very active, the discovery of leptin moved the status of adipose tissue towards an endocrine tissue able to interact with all major organs via secretion of adipokines. Some years ago, the presence of adipocyte precursors, termed preadipocytes, has been described in all adipose tissue depots from various species of different age. More recently, the discovery that different phenotypes can be obtained from stroma cells of adipose tissue has largely emphazised the concept of adipose tissue plasticity. Therefore, raising great hope in regenerative medicine as adipose tissue can be easily harvested in adults it could represent an abundant source of therapeutic cells. Thus, adipose tissue plays the dual role of Mr Obese Hyde as a main actor of obesity and of Dr Regenerative Jekyll as a source of therapeutic cells. Adipose tissue has not yet revealed all its mysteries although one facet could not be well understood without the other one. PMID- 17110895 TI - Diabetes mellitus and dementia. AB - Alzheimer's disease (AD) and diabetes mellitus (DM) are two of the most common and devastating health problems in the elderly. They share a number of common features amongst which high prevalence after 65 years, important impact of patient's quality of life, substantial health care costs. Reviews on the epidemiological studies on cognitive impairment in patients with DM found evidence of cross-sectional and prospective associations between type 2 DM and moderate cognitive impairment, on memory and executive functions. There is also evidence for an elevated risk of both vascular dementia and AD in patients with type 2 DM, albeit with strong interaction of other factors such as hypertension, dyslipidaemia and ApoE genotype. DM is an independent predictor of post-stroke dementia. DM being an atherogenic risk factor, it may increase the risk of dementia through associations with stroke, causing vascular dementia. In addition, vascular reactivity may be adversely affected by advanced glycosylation end products resulting in more subtle perfusion abnormalities. Cerebrovascular disease may exacerbate AD through direct interactions between the two pathological processes or through cognitive impairment secondary to cerebrovascular disease "unmasking" AD at an earlier stage than it would otherwise become apparent. The increased risk of AD may also be mediated by the exacerbation of B-amyloid neurotoxicity by advanced glycosylation end products identified in the matrix of neurofibrillary tangles and amyloid plaques in AD brains, or associations with insulin functions. Decreased cholinergic transport across the blood-brain barrier observed in diabetic animals may exacerbate cognitive impairment in AD. Many interventions could reduce the cognitive decline associated with DM, yet not enough are taken into account so far. PMID- 17110896 TI - The role of alexithymia factors in glucose control of persons with type 1 diabetes: a pilot study. AB - OBJECTIVE: To clarify the respective contribution of demographic characteristics, health conditions and three psychological variables (depression, anxiety, alexithymia) for glycaemic control measured by glycated hemoglobin (HbA1c). MATERIALS AND METHODS: Sixty-four persons diagnosed with type 1 diabetes completed psychological measures and demographic information at admission (T1) to the hospital and in a follow-up (+8 weeks) (T2). Additional information about their health conditions was also considered. RESULTS: At T1, the alexithymia factor "difficulties describing feelings" (DDF) predicted HbA1c over and above the predictive power of demographic information, health conditions, anxiety, and depression. Additionally, higher decrease in HbA1c from T1 to T2 was predicted by higher scores on the alexithymia factor DDF at admission over and above the other predictors. CONCLUSION: DDF is an important predictor of glucose control. Scoring higher on this factor is related to poorer glycaemic control at admission. Additionally, people with higher scores on this factor seem to benefit highly from the treatment administered at the hospital. PMID- 17110897 TI - Continuous intraperitoneal insulin infusion does not increase the risk of organ specific autoimmune disease in type 1 diabetic patients: results of a multicentric, comparative study. AB - AIM: The purpose of this national multicenter prospective study by the French EVADIAC group was to investigate the possibility that continuous intraperitoneal insulin infusion using an implanted pump (CIpii) increases the risk of autoimmune disease in type 1 diabetic patients as it increased anti-insulin immunogenicity. METHODS: Prevalence of clinical (Hashimoto's disease, hyperthyroidism, gastric atrophic disease and vitiligo) and subclinical (presence of anti-thyroperoxidase antibodies, anti-intrinsic factor antibodies, abnormal TSH levels) autoimmune diseases was estimated by comparing two groups of patients already treated by either CIpii (n=154) or external pump (CSII) (n=121) for an average of 6 years. Incidence of autoimmune disease was determined by comparing the same measurements one year after inclusion. RESULTS: No significant difference was observed for the total prevalence of clinical and subclinical auto-immune thyroid and gastric di seases (35.6% and 3.2% respectively in the CIpii group versus 40.4% and 2.6% in the CSII group). No significant difference for the incidence of clinical and subclinical auto-immune diseases was observed: 7.2% and 0% in CIpii and 7.3% and 1.7% in CSII. CONCLUSION: As previously shown AIA (anti-insulin antibodies) levels were higher in CIpii than in CSII (32.9% vs 20.2%, P<0.0001) but no correlation was observed with either clinical or subclinical autoimmune disease. This large-scale study eliminates the possibility that CIpii increases the risk of autoimmune disease. PMID- 17110898 TI - Difficulties of smoking cessation in diabetic inpatients benefiting from a systematic consultation to help them to give up smoking. AB - AIM: To assess the value of systematic smoking cessation consultations for diabetic smokers admitted to hospital. METHODS: All diabetic smokers admitted to the Diabetes Department of Georges Pompidou European Hospital between February 2003 and February 2004 were systematically offered a consultation with a physician specialised in tobacco cessation. Follow-up visits at three, six and nine months were planned. RESULTS: Of the 306 diabetic patients admitted, 38 (12.4%) were smokers. There were more men than women in the group of smokers and the diabetic smokers were younger than the non-smokers. The smokers had fewer micro-angiopathic complications than the non-smokers, but there was no difference in the frequency of macro-angiopathic complications. The level of nicotine physical dependence was moderate or high for 60% of the smokers. Although all the smokers agreed to the consultation, less than half agreed to drug-based treatments to help them to give up smoking and only 15% returned for the six month visit. Only one patient had stopped smoking at the six-month visit. CONCLUSION: This study demonstrates the difficulties in systematic interventions to help diabetic patients to stop smoking. Diabetic smokers probably constitute a specific population for which the barriers to giving up smoking should be explored. PMID- 17110899 TI - Glucose intolerance and other cardiovascular risk factors in Haiti. Prevalence of Diabetes and Hypertension in Haiti (PREDIAH). AB - AIMS: To assess the prevalence of diabetes and other forms of glucose intolerance and to examine their relationship with some cardiovascular risk factors in a population representative of the capital of Haiti. METHODS: This cross-sectional survey was conducted in the metropolitan area of Port-au-Prince, Haiti. A population-based sample of 1620 adults was randomly selected using a two-stage cluster method, stratified in 3 age groups: 20-39, 40-64 and > or =65 years of age. Diagnosis of diabetes and pre-diabetes (IFG and IGT) was based on the 2003 Expert Committee criteria. RESULTS: The total response rate was 69%. The age standardized prevalence of diabetes was 4.8% in men and 8.9% in women (P=0.0014), with, overall, 70.6% of previously diagnosed cases. Standardized for the Segi world population aged 30-64 years, its prevalence was 7.4% in men and 11.1% in women (NS). The age-standardized prevalence of pre-diabetes was 6.4% in men and 8.0% in women (NS). Hypertension was found in 48.7% in men and 46.5% in women (NS). Its rates in people > or =40 years old were 69.1% in men and 67.2% in women (NS). Abdominal obesity was strongly independently associated with diabetes and pre-diabetes in both genders. Hypertension was shown to be risk factor for pre diabetes and total glucose intolerance in women. High education was associated with lower risk of diabetes in men. CONCLUSION: Prevalence of diabetes and pre diabetes is moderately high in Port-au-Prince, Haiti. In people aged > or =40 years, the rate of normal blood pressure is less than 25%. Intervention programs to prevent simultaneously and manage diabetes and hypertension are imperative, and prevention strategies through lifestyle modifications should be cost effective. PMID- 17110900 TI - Bone density and markers of bone remodeling in type 1 male diabetic patients. AB - AIMS: To assess the prevalence and severity of bone disease in type 1 diabetic patients and to determine serum markers of bone remodeling as well as their relationship with bone mineral density (BMD). METHODS: BMD [by dual energy x-ray absorptiometry (DXA)] and serum markers of bone remodeling [osteocalcin, c terminal telopeptide of type I collagen (CTX)], leptin and osteoprotegerin (OPG) were measured in 42 adult males with type 1 diabetes. Twenty-four non-diabetic subjects served as controls. RESULTS: In 40% of the patients, osteopenia at the lumbar spine (L1-L4) and/or at the left hip was found, and 7% met criteria for osteoporosis. L1-L4 BMD z-score was correlated with age (r=0.365, P=0.018) and a similar trend was observed at left hip. L1-L4 BMD z-score was negatively correlated with CTX and osteocalcin (r=-0.343, P=0.028; r=-0.376, P=0.024, respectively). A significant correlation was evidenced between BMD z-score at both lumbar spine and left hip and leptin values (r=0.343, P=0.03; r=0.395, P=0.012, respectively) but after adjustment for weight this correlation was no longer significant. Osteocalcin, CTX and leptin concentrations were comparable between patients and controls, while OPG concentrations tend to be higher in diabetic subjects (P=0.08). CTX was negatively correlated with age (r=-0.390, P=0.012) and positively correlated with osteocalcin (r=0.696, P<0.001). OPG was positively correlated with age (r=0.507, P=0.001). CONCLUSION: Our results suggest that in diabetic subjects osteopenia is a relatively frequent complication but bone loss is attenuated with age progression. Whether this is also mediated by OPG and/or leptin remains to be confirmed. PMID- 17110901 TI - Characteristics and six-month outcomes in a cohort of 8288 diabetic and non diabetic patients with previous history of acute coronary syndrome or stroke: the French PREVENIR 3 survey. AB - AIMS: To evaluate the cardiovascular prognosis of 1845 Diabetic Patients (DP) and 6443 Non-Diabetic Patients (NDP) in secondary prevention. METHODS: Patients were recruited prospectively if they had had a previous history of ischemic stroke or acute coronary syndrome (ACS) i.e. Myocardial Infarction (MI) or Unstable Angina (UA) within a period of five years preceding inclusion. For each patient, the number of hospitalizations and vital status were recorded each month over a 6 month period (mean follow-up: 4.8 months). RESULTS: 306 patients (9.5/100--person years; 95% CI, 8.5 to 10.6) had undergone at least one subsequent event (hospitalization for ACS, ischemic stroke, or cardiovascular death). A majority of these events were non-fatal ACS (n=248). The cumulative incidence rate of subsequent events was higher in DP: 12.6/100- person years (10.0 to 15.2) than in NDP: 8.6/100--person years (7.5 to 9.8). DP were significantly at higher risk of subsequent cardiovascular events (OR: 1.34; P=0.025) after adjustment for confounding factors. 93% of coronary DP and NDP underwent a recurrent event affecting the same location. When the index episode was a stroke, 71% of DP had a subsequent stroke vs. 47% of NDP. CONCLUSION: in secondary prevention, the risk of mortality and subsequent vascular events is independently higher in French DP than in NDP. The locations affected by each type of subsequent cardiovascular event seemed correlated to the baseline diagnosis, whatever the diabetic status, even when the frequency of subsequent strokes increased (not significantly) in DP when compared to NDP. PMID- 17110902 TI - Cardiovascular risk markers associated with the metabolic syndrome in a large French population: the "SYMFONIE" study. AB - OBJECTIVE: The SYMFONIE study was designed to analyze the clinical and biological characteristics, and the cardiovascular risk markers, in men and women with the metabolic syndrome compared to control subjects. RESEARCH DESIGN AND METHODS: The study population included 101,697 men and women, 18 to 80 years of age, who had a health checkup at the Centre d'Investigations Preventives et Cliniques (Paris, France) between 1997 to 2002. The metabolic syndrome was defined according to the ATpiiI-NCEP 2001 criteria. RESULTS: Out of the 66,202 men (47.4+/-11.8 years) and 35,495 women (48.5+/-13.6 years) included in this population, 6761 men (10.2%) and 2155 women (6.1%) presented the metabolic syndrome. Among subjects < or =40 years of age, the prevalence of the metabolic syndrome was 5.0% in men and 2.2% in women, and rose to 14.1% and 12.0%, respectively, among men and women >70 years of age. After adjustment for age, patients with the metabolic syndrome presented higher pulse pressure (systolic minus diastolic blood pressure), higher heart rate, lower vital respiratory capacity, lower physical activity, an increase in inflammatory status assessed through leukocyte count and dental inflammation, hepatic abnormalities, and increased levels of stress and depression. CONCLUSION: In this large French population, the prevalence of the metabolic syndrome is lower than in North American and northern European populations. Patients with the metabolic syndrome present several additional hemodynamic, inflammatory and psychological risk markers which could contribute to the poor cardiovascular prognosis of these subjects. PMID- 17110903 TI - Metabolic syndrome and coronary heart disease risk in a population-based study of middle-aged men from France and Northern Ireland. A nested case-control study from the PRIME cohort. AB - Metabolic Syndrome (MetS) was found associated with an increased CHD risk in several studies but data about this relationship in Southern Europe are lacking. We studied the association of MetS according to three different indexes (the National Cholesterol Education Program's definition (NCEP), a modified World Health Organization's definition (WHO) and the recent International Diabetes Federation's definition (IDF)) with CHD risk in a case-control study nested within the PRIME cohort, composed of subjects from France (Southern Europe) and Belfast (Northern Europe). The PRIME prospective study is composed of 10 592 men, aged 50-59 at baseline and followed for 5 years. Subjects included in this nested case-control study were 296 cases of incident CHD and 540 controls, who remained free of CHD during the 5 years of follow-up of the PRIME cohort and matched for age, recruitment centre and recruitment date. All subjects had questionnaires and a medical examination at baseline, and a blood sample was taken. Using the IDF's, the WHO's and the NCEP's definitions respectively, the frequency of MetS was 38.9%, 35.5% and 29.7% in cases and 32.4%, 28.7% and 22.6% in controls. After adjustment for physical activity, smoking and drinking habits, MetS was associated with CHD risk whichever the definition used (ORIDF=1.41 [1.02-1.95], P<0.04, ORWHO=1.40 [1.01-1.94], P<0.05 and ORNCEP=1.46[1.04-2.04], P<0.04). These results were homogeneous in France (low risk of CHD) and Belfast (high risk of CHD). Our results add further evidence that MetS is predictive of CHD risk in middle-aged men from Northern and Southern Europe, and highlight differences between the three definitions studied. PMID- 17110904 TI - Scleredema adultorum of Buschke: an under recognized skin complication of diabetes. AB - Scleredema of Buschke or scleredema diabetorum is a skin complication of diabetes with deposits of collagen and aminoglycans in the dermis. This disease characterized by thickening and hardening of the skin, is usually localized in nape, back and shoulder areas. Consequences could be a decrease in motility of the shoulders and an impairment of respiratory function. Other possible complications are sleep apnoea syndrome and monoclonal gammapathy. Type 1 or type 2 diabetes may be associated with scleredema of Buschke in more than 50% of cases. Diabetes-related risk factors are long duration of the disease, presence of microangiopathy, overweight and need of insulin. Various specific treatments proposed in the literature are poorly validated. In most severe cases, radiation therapy may be useful. PMID- 17110905 TI - Smoking is associated with increased levels of osteopontin in type 2 diabetic patients: preliminary results. PMID- 17110906 TI - Vitronectin is present in epithelial cells of the intact lens and promotes epithelial mesenchymal transition in lens epithelial explants. AB - PURPOSE: Extracellular matrix (ECM) accumulates during the development of posterior capsule opacification (PCO). Vitronectin, an ECM component that is generally prominent in wound healing, has been detected in PCO specimens. Here we set out to investigate the distribution of vitronectin in the lens and determine how it, and other ECM components, influence the lens epithelial phenotype. METHODS: Rat lens epithelial explants were cultured on vitronectin, fibronectin, and laminin substrata. Explants were monitored for cell migration and the appearance of markers for epithelial mesenchymal transition (EMT), using phase contrast microscopy and immunohistochemistry, respectively. Explants were also monitored for evidence of Smad signaling. Vitronectin expression was analyzed in embryonic and postnatal rodent lens development by immunohistochemistry, western blotting, and in situ hybridization. RESULTS: Vitronectin, like fibronectin and laminin, provided a good substratum for cellular attachment and migration. However, in the case of vitronectin and fibronectin, this was accompanied by a major phenotypic change. On either vitronectin or fibronectin, but not laminin, most of the cells became elongated, spindle-shaped and were strongly reactive for filamentous alpha-smooth muscle actin. In these respects this transition was typical of the well known TGFbeta-induced EMT. In explants cultured on vitronectin and fibronectin, but not laminin, cell nuclei showed prominent reactivity for Smad 2/3. Vitronectin was also shown to be expressed during embryonic and postnatal development. Initially mRNA and protein were detected in all lens cells, however as development progressed, expression became restricted to cells of the epithelium and transition zone. CONCLUSIONS: The results clearly show that lens cell engagement with a vitronectin or a fibronectin, but not laminin, substratum has a potent EMT promoting effect and that Smad 2/3 signaling is involved. Thus when considering strategies to slow or prevent PCO, these results highlight the need to take into account ECM molecules such as vitronectin that have the capacity to promote EMT. PMID- 17110907 TI - Inhibitory effect of an antibody to cryptic collagen type IV epitopes on choroidal neovascularization. AB - PURPOSE: The wet form of age-related macular degeneration (AMD) occurs as a consequence of abnormal blood vessel growth from the choroid into the retina. Pathological angiogenesis during tumor growth and ocular disease has been associated with specific exposure of cryptic extracellular matrix epitopes. We investigated the presence of cryptic collagen IV epitopes in a murine model of choroidal neovascularization (CNV), and tested the effect on blood vessel growth of H8, a humanized antibody directed against a cryptic collagen type IV epitope. METHODS: To induce experimental CNV in adult C57BL/6 mice, Bruch's membrane was ruptured using a diode laser. Subsequently, mice were treated with daily intraperitoneal (i.p.) injections of either H8 (10 mg/kg or 30 mg/kg) or an isotype-matched antibody control. Two weeks postinjection, choroidal flat mounts were immunostained with the blood vessel marker platelet/endothelial cell adhesion molecule-1 (PECAM-1) and H8. CNV was visualized using fluorescence microscopy and the CNV lesion area measured using Open Lab software. RESULTS: Collagen type IV and the cryptic epitope were observed at the site of laser induced lesions. Staining with H8 was first observed three days post injury, two days after MMP2 expression in CNV lesions, becoming most intense five days following laser injury and extending beyond the area of neovascularization. At 14 days post injury, H8 staining was reduced in intensity, colocalized with the area of CNV, and was nearly absent from the underlying choroidal vessels. In addition, mice treated with H8 had a significant dose-dependent decrease in the area of CNV as compared to isotype-matched antibody controls. CONCLUSIONS: Results suggest that exposure of cryptic collagen type IV epitopes is associated with the incidence of CNV and that the humanized antibody H8 may provide a new treatment for CNV. PMID- 17110908 TI - Egr1 gene knockdown affects embryonic ocular development in zebrafish. AB - PURPOSE: To identify the changes in zebrafish embryonic ocular development after early growth response factor 1 (Egr1) gene knockdown by Egr1-specific translation inhibitor, morpholino oligonucleotides (MO). METHODS: Two kinds of Egr1-MO were microinjected separately with various dosages into one to four celled zebrafish embryos to find an optimal dose generating an acceptable mortality rate and high frequency of specific phenotype. Chordin-MO served as the positive control; a 5 mismatch MO of Egr1-MO1 and a nonspecific MO served as negative controls. We graded the Egr1 morphants according to their gross abnormalities, and measured their ocular dimensions accordingly. Western blot analysis and synthetic Egr1 mRNA rescue experiments confirmed whether the deformities were caused by Egr1 gene knockdown. Histological examination and three kinds of immunohistochemical staining were applied to identify glutamate receptor one expression in retinal ganglion cells and amacrine cells, to recognize acetylated alpha-tubulin expression which indicated axonogenesis, and to label photoreceptor cells with zpr-1 antibody. RESULTS: After microinjection of 8 ng Egr1-MO1 or 2 ng Egr1-MO2, 81.8% and 97.3% of larvae at 72 h postfertilization had specific defects, respectively. The gross phenotype included string-like heart, flat head, and deformed tail. The more severely deformed larvae had smaller eyes and pupils. Co injection of 8 ng Egr1-MO1 and supplementary 12 pg synthetic Egr1 mRNA reduced the gross abnormality rate from 84.4% to 29.7%, and decreased the severity of deformities. Egr1 protein appeared in the wildtype and rescued morphants, but was lacking in the Egr1 morphants with specific deformities. Lenses of Egr1 morphants were smaller and had some residual nucleated lens fiber cells. Morphants' retinal cells arranged disorderly and compactly with thin plexiform layers. Immunohistochemical studies showed that morphants had a markedly decreased number of mature retinal ganglion cells, amacrine cells, and photoreceptor cells. Retinal axonogenesis was prominently reduced in morphants. CONCLUSIONS: The Egr1 gene plays an important role in zebrafish embryonic oculogenesis. Ocular structures including lens and retina were primitive and lacked appropriate differentiation. Such arrested retinal and lenticular development in Egr1 morphants resulted in microphthalmos. PMID- 17110909 TI - Pre-mRNA splicing and retinitis pigmentosa. AB - Retinitis pigmentosa (RP) is a group of genetically and clinically heterogeneous retinal diseases and a common cause of blindness. Among the 12 autosomal dominant RP (adRP) genes identified, four encode ubiquitously expressed proteins involved in pre-mRNA splicing, demonstrating the important role that pre-mRNA splicing plays in the pathogenesis of retinal degeneration. This review focuses on recent progress in identifying adRP mutations in genes encoding pre-mRNA splicing factors and the potential underlying molecular mechanisms. PMID- 17110910 TI - Decrease of cone opsin mRNA in experimental ocular hypertension. AB - PURPOSE: This study was designed to test the hypothesis that photoreceptors are adversely affected in glaucoma. As a measure of this effect, we examined the levels of rod opsin, and red/green and blue cone opsin mRNAs in monkeys with experimental ocular hypertension and glaucoma and in human eyes from donors with diagnosed glaucoma. METHODS: Experimental ocular hypertension was induced in one eye of 19 cynomolgous and 2 rhesus monkeys by laser ablation of the trabecular meshwork. In 15 monkeys, the elevated IOP was reduced by trabeculectomy. When the animals had experienced prolonged elevations of IOP (128 to 260 days), they were killed and the eyes enucleated. Fresh retinal tissue from the macula, inferotemporal retina (mid-peripheral), and far peripheral regions were harvested from some animals using a 3 mm trephine. The remaining retinas from these monkeys, and whole retinas from other animals were fixed. RNA isolated from each trephined sample was used for RNase Protection Analysis or real time PCR analysis to quantify opsin mRNA levels from different photoreceptor cell types. Fixed tissue was used for in situ hybridization studies. Human donor eyes (7 glaucoma and 4 control) were obtained from eye banks. All human specimens were used for in situ hybridization studies. RESULTS: Quantitative mRNA analysis and in situ hybridization studies both showed a reduction in the expression of red/green and blue cone opsin mRNAs in 6 monkey eyes with chronic ocular hypertension, relative to the contralateral eye. No loss of rod opsin mRNA was observed. The principal reduction occurred in cells of the mid-peripheral retina, a region of retina that often shows early and progressive damage in humans with glaucoma. In monkeys with ocular hypertension followed by trabeculectomy, there was a similar decrease in cone opsin mRNAs, but only in six out of fifteen (40%) of the monkeys. The decrease in these animals was correlated with a significantly elevated IOP at some time during the 2 weeks prior to euthanization and not with the extent of glaucomatous damage. Of the 7 human eyes with diagnosed glaucoma that were examined, 5 showed a decrease of cone opsin mRNA in the mid-peripheral retina, whereas none of the 4 normal eyes examined showed a decrease. CONCLUSIONS: Ocular hypertension leading to glaucoma also affects the outer retina, particularly the cone photoreceptors. We speculate that these cells become stressed leading to a disruption in the expression of normal genes, such as that encoding opsin. There is some evidence that this effect is reversible, when IOP levels are reduced. PMID- 17110911 TI - Mutations in the gene encoding the alpha-subunit of rod phosphodiesterase in consanguineous Pakistani families. AB - PURPOSE: To localize and identify the gene and mutations causing autosomal recessive retinitis pigmentosa (RP) in consanguineous Pakistani families. METHODS: Families were ascertained and patients underwent complete ophthalmological examinations. Blood samples were collected and DNA was extracted. A genome-wide scan was performed using 382 polymorphic microsatellite markers on genomic DNA from affected and unaffected family members, and lod scores were calculated. RESULTS: A genome-wide scan of 50 families gave a lod score of 7.4172 with D5S2015 using HOMOG1. RP in all 4 linked families mapped to a 13.85 cM (14.87 Mb) region on chromosome 5q31-33 flanked by D5S2090 and D5S422. This region harbors the PDE6A gene, which is known to cause autosomal recessive RP. Sequencing of PDE6A showed a homozygous single base pair change; c.889C->T, single base pair insertion; c.2218-2219insT, and single base pair substitution in the splice acceptor site; IVS10-2A->G in each of three families. In the fourth family linked to this region, no disease-causing mutation was identified in the PDE6A gene. CONCLUSIONS: These results provide strong evidence that mutations in PDE6A result in recessive RP in three consanguineous Pakistani families. Although a fourth family was linked to markers in the 5q31-33 interval, no mutation was identified in PDE6A. PMID- 17110912 TI - The rat Apg3p/Aut1p homolog is upregulated by ischemic preconditioning in the retina. AB - PURPOSE: Retinas can be protected from subsequent severe ischemic injury by ischemic preconditioning. Ischemic preconditioning is dependent on gene expression and protein synthesis; however, it is not clear which genes are important in this process. In this study, we have identified and characterized the rat homolog of yeast Apg3p/Aut1p, an important autophagy protein encoded by the autophagy 3-like (APG3L) gene. We have also further characterized the homologous human APG3L gene. METHODS: A fragment of the rat Apg3 cDNA was identified by mRNA differential display from hypoxia-treated E1A-NR3, an immortalized cell line derived from rat retinal cells that manifests phenotypes of retinal neurons. The full length of rat Apg3 (rApg3) cDNA sequence (about 1.4 kb) encoding 341 amino acids was cloned from a rat retinal cDNA library and characterized using Southern and northern blot analysis, and a global GenBank search. Protein expression was determined by western blotting, and immunohistochemistry. Ischemic preconditioning was achieved by ligation of the retinal arteries of the right eye for 5 min followed by 5 h reperfusion. The prolonged retinal ischemia was induced by ligation of the retinal arteries for 45 min followed by 5 h reperfusion. The full-length homologous human APG3L gene was cloned and sequenced from a human genomic DNA library. RESULTS: The combination of genomic Southern blot analysis and a global GenBank search indicated that rat APG3L is a single copy gene. Rat Apg3 mRNA is expressed in the retina at a high level but is also detected in other tissues. In the process of comparing the rat and human APG3L genes we showed that the organization of the human APG3L gene includes a unique transcriptional start site, a coding region with 12 translated exons and 11 introns and is located on human chromosome 3q13.1. Subcellular localization studies showed that recombinant rat autophagocytosis protein (Apg3p) is a cytosolic protein. Rat Apg3 mRNA level was upregulated by ischemic preconditioning but downregulated by prolonged ischemia. CONCLUSIONS: Our results suggest that the upregulation of rApg3 is a specific response to ischemic preconditioning rather than to retina ischemia, and autophagy may contribute to the neuroprotective effect of ischemic preconditioning in the retina. PMID- 17110913 TI - Intravital microscopy of leukocyte-endothelial dynamics using the Heidelberg confocal laser microscope in scleritis and allergic conjunctivitis. AB - PURPOSE: To examine leukocyte-endothelial cell rolling and arrest in human ocular vessels overlying sites of inflammation in various ocular inflammatory diseases in comparison to normal controls using the Heidelberg confocal laser microscope, which provides images with greater clarity and resolution than the tandem scanning microscope that uses white light. METHODS: Healthy controls (n=8) and patients with active anterior scleritis (n=7) or allergic eye disease (n=4) were scanned using the Heidelberg confocal laser microscope (HRT II) with the Rostock cornea module attachment for a minimum of 5 min at a depth of 45-120 microm from the conjunctival epithelial surface. RESULTS: There was a marked increase in the number of rolling leukocytes in scleritis patients (534+/-119 cells per mm2/min) versus controls (6+/-6 cells per mm2/min; p=0.0002) or allergic patients (59+/-44 cells per mm2/min; p=0.009). No statistically significant increase was seen in allergic patients compared to controls (p=0.059). A similar pattern was seen in the number of arrested leukocytes in patients with scleritis (56+/-23 cells per mm2) in comparison to either those with allergic eye disease or controls (each=0 cells per mm2; p=0.02). CONCLUSIONS: Patients with scleritis have a significantly increased number of rolling and arrested leukocytes in superficial ocular vessels in comparison to patients with mild allergic conjunctivitis and controls. The image quality with this microscope is superior to prior studies with a scanning microscope. PMID- 17110914 TI - Uptake of cholesterol by the retina occurs primarily via a low density lipoprotein receptor-mediated process. AB - PURPOSE: In this study we examined the uptake of circulating lipoproteins into the retina, using a naturally fluorescent cholesterol analog for imaging and deuterated cholesterol for quantification by mass spectroscopy. The purpose of this study was to better understand cholesterol uptake, transport and homeostasis in the retina. METHODS: Human low density lipoprotein (LDL) and high density lipoprotein (HDL) were labeled with the fluorescent cholesterol analog cholesta 5,7,9(11)-trien-3beta-ol (CTL) and deuterated cholesterol (25,26,26,26,27,27,27 [2H]cholesterol, D7Ch). Rats were injected intravenously with CTL-LDL, CTL-HDL and D7Ch-LDL. Fluorescent confocal microscopy was used to image the uptake of CTL and mass spectroscopy was used to quantify D7Ch. Immunohistochemistry and fluorescent confocal microscopy were used to localize apoB (an LDL marker protein) and LDL receptor (LDLR) protein in rat and monkey retinas. RESULTS: CTL specific fluorescence was imaged by confocal microscopy in the retinal pigment epithelium (RPE), choriocapillaris and parts of the neural retina within 2 h post injection and was visualized in the photoreceptor outer segments by 4 h. Replacing LDL with HDL as the CTL carrier gave a less robust and more delayed labeling of retinal layers. Human apolipoprotein B (apoB) was also localized in the rat choriocapillaris and RPE by 4 h post-injection. Human apoB was detected by immunoblot analysis in the rat retina primarily as a about 70 kDa protein, suggesting proteolytic degradation. LDL-mediated uptake of cholesterol was quantified by mass spectroscopy using deuterated cholesterol in place of CTL. In addition, apoB and LDLR were localized in monkey retina by immunohistochemistry. CONCLUSIONS: The retina is capable of rapid uptake of circulating LDL via an LDLR mediated process primarily occurring in the RPE and also possibly Muller cells. Despite the dominance of HDL over LDL in rat serum, LDL appears to be the preferred carrier for cholesterol transport to and uptake by the retina. The results also suggest that blood-borne LDL represents a significant contributor to the steady-state levels of cholesterol and possibly other lipids in the retina. PMID- 17110915 TI - Intraretinal lipid transport is dependent on high density lipoprotein-like particles and class B scavenger receptors. AB - PURPOSE: In our companion paper we demonstrated that circulating lipoproteins enter the retina via the retinal pigment epithelium (RPE) and possibly Muller cells. In order to understand how these lipids are transported within the retina, expression and localization of the main proteins known to be involved in systemic lipid transport was determined. METHODS: Expression of ABCA1, apoA1 (the major HDL protein), SR-BI, SR-BII, CD36, lecithin:cholesterol acyltransferase (LCAT), and cholesteryl ester transfer protein (CETP) was determined by reverse transcriptase polymerase chain reaction (RT-PCR) and immunoblots. Localization was determined by immunohistochemistry using fresh monkey vibrotome sections and imaged by confocal microscopy. RESULTS: ABCA1 and apoA1 were localized to the ganglion cell layer, retinal pigment epithelium (RPE), and rod photoreceptor inner segments. ApoA1 was also observed associated with rod photoreceptor outer segments, presumably localized to the interphotoreceptor matrix (IPM). The scavenger receptors SR-BI and SR-BII localized mainly to the ganglion cell layer and photoreceptor outer segments; in the latter they appear to be associated with microtubules. LCAT and CETP localized mainly to the IPM. CONCLUSIONS: The presence and specific localization of these well-known lipid transport proteins suggest that the retina employs an internal lipid transport mechanism that involves processing and maturation of HDL-like particles. PMID- 17110916 TI - Ocular gene therapy: a review of nonviral strategies. AB - Along with viral vectors, non-viral strategies have been developed in order to efficiently deliver nucleic acids to ocular cells. During the last decade, we have observed that the outcome of these non-viral delivery systems depends on the genetic material used, the targeted tissue or cells, the expected effect duration, and the routes of administration. Assessment of efficiency has been evaluated in normal eyes or in animal models of ocular diseases. The chemical and physical methods that have been adapted for the delivery of nucleic acids to ocular tissues are highlighted and discussed in this review. Also, the results obtained with different non-viral strategies from their initial conception to their present development are summarized. At the present, selective targeting of ocular tissues and cells can be achieved using the most yielding route of administration to the eye in combination with an appropriate drug delivery technique. PMID- 17110917 TI - Retinal pigment epithelium cell damage by A2-E and its photo-derivatives. AB - PURPOSE: A2-E, a major component of the retinal pigment epithelium (RPE) lipofuscin, is a compound that can neither be degraded by nor eliminated from cells and is toxic as well as phototoxic to the cells. Illumination of A2-E with short wavelength light results in isomerization, photooxidation, as well as photolysis. Cytotoxic intermediates (free oxygen radicals) and reaction products (peroxides) are involved in this process. METHODS: A2-E solution (1.28 mM in ethanol or 10 microM phosphate-buffered saline) was kept in dark, exposed to blue light (450-490 nm, 0.2 mW/mm2) for 15 min, or to white light (8.9 mW/mm2) for 60 min, respectively and supplemented to the culture medium of primary porcine RPE cells for 24 h. Damaged cells were determined by staining with propidium iodide in 24 experiments. The photooxidation products of A2-E were analyzed by ultraviolet-visible spectroscopy and MALDI-TOF mass spectrometry. RESULTS: Supplementation of A2-E for 24 h resulted in a rate of damaged cells of 28%. Blue light exposure of A2-E before supplementation increased the rate to 91% whereas the exposure to high dosage white light reduced it to 14%. Irradiation of A2-E resulted in a dosage-dependent addition of one through four oxygen atoms. CONCLUSIONS: The increase of the cell damage rate by A2-E irradiated with low dosage light supports the hypothesis of direct DNA damage by oxidized A2-E. Furthermore, we found a reduced cell damage rate from intensively irradiated A2-E resulting in a tetraoxidized molecule which was rather stable and thus less toxic. PMID- 17110918 TI - Fenretinide-induced neuronal differentiation of ARPE-19 human retinal pigment epithelial cells is associated with the differential expression of Hsp70, 14-3-3, pax-6, tubulin beta-III, NSE, and bag-1 proteins. AB - PURPOSE: We reported earlier that fenretinide can induce neuronal differentiation of ARPE-19 human retinal pigment epithelial cells in culture. The purpose of this study was to investigate the potential involvement of key proteins involved in gene transcription, signal transduction, cell cycle check point, differentiation, neuronal cell survival, and stress response in the neuronal differentiation of ARPE-19 cells by fenretinide. METHODS: Cells in culture were treated with 1.0 microM fenretinide. Cells were analyzed using antibodies against pax-6, neuronal specific enolase (NSE), tubulin beta-III, 14-3-3, bag-1, and Hsp-70 proteins using immunocytochemistry, western blot and ELISA methodologies. RESULTS: We found that pax-6 and NSE were both expressed in the control ARPE-19 cells. Fenretinide induced neuronal differentiation of ARPE-19 cells led to a decrease in pax-6 protein and an increase in tubulin beta-III protein expression after 5 days fenretinide treatment. There was a translocation of 14-3-3 from the cytoplasm to the nucleus, and an increase in nuclear expression of bag-1 after treatment. We also found a time-dependent increase in Hsp70 protein expression in ARPE-19 cells treated with fenretinide. D-407, another human retinal pigment epithelial cell line, but not either Y-79 or PC-12 cells, was also able to be induced into neuronal morphologies by fenretinide. CONCLUSIONS: The fenretinide induced neuronal differentiation of ARPE-19 cells is associated with an increase in expression of the neuronal specific protein tubulin beta-III, and a decrease in expression of the progenitor cell marker pax-6. Neuronal differentiation of ARPE-19 cells is also associated with nuclear translocation of 14-3-3, a protein involved in signal transduction, cell cycle check point and cell growth, and an increase in expression of bag-1, a protein involved in neuronal cell survival and axon elongation. These results suggest that ARPE-19 cells could be a progenitor cell line that can be differentiated into neuronal cells when treated with factors such as fenretinide. PMID- 17110919 TI - The association of single nucleotide polymorphisms in the MMP-9 genes with susceptibility to acute primary angle closure glaucoma in Taiwanese patients. AB - PURPOSE: To study the relationships between single nucleotide polymorphisms (SNPs) of extracellular matrix, matrix metalloproteases (MMPs), tissue inhibitors of MMPs, and other glaucoma-associated genes and acute primary angle closure glaucoma (PACG). METHODS: We extracted DNA samples from 78 adult patients with acute PACG and 86 control subjects to study the relationships between these specific genes and acute PACG. Genotyping was performed for 35 genes by the GenomeLab SNPstream genotyping system after PCR amplification of chromosomal DNA. The association between these genetic polymorphisms and risk of primary PACG was estimated by chi2 and logistic regression. RESULTS: The genotyping success rate was 99%. Genotyping for the MMP9 site (rs2664538) was significantly different between the two groups (p=0.000001) and the odds ratio was 2.586 (95% CI: 1.715 3.898, p<0.00001). However, there were no associations of SNPs to other genes in patients with acute PACG. CONCLUSIONS: Our results reveal that SNP rs2664538, which is located at the MMP9 gene, is likely to be associated with acute PACG. PMID- 17110920 TI - A novel mutation in GJA8 associated with autosomal dominant congenital cataract in a family of Indian origin. AB - PURPOSE: To identify the genetic defect in an autosomal dominant congenital cataract family, having 15 members in three generations, affected with bilateral cataract that gave the appearance of "full moon" with Y-sutural opacities. METHODS: A detailed family history and clinical data were recorded. A genome-wide scan by two point linkage analysis using nearly 400 microsatellite markers in combination with multipoint lod score and haplotype analysis was carried out. Mutation screening was performed in the candidate gene by bidirectional sequencing of amplified products. RESULTS: A maximum two point lod score of 5.45 at theta=0.00 was obtained with marker D1S534. Haplotype analysis placed the cataract locus to a 14.1 cM region between D1S221 and D1S498, in close proximity to the gene for the gap junction channel protein connexin 50 (GJA8) at 1q21. Mutation screening in GJA8 identified a novel G>C transversion at nucleotide position c.235. This nucleotide change resulted in the substitution of highly conserved valine by leucine at codon 79 (V79L). This nucleotide substitution was neither seen in any unaffected member of the family nor in 180 unrelated control subjects (360 chromosomes) from same ethnic background tested by sequence analysis of GJA8. CONCLUSIONS: The present study describes the mapping of a locus for congenital cataract that appeared like "full moon" with Y-sutural opacities at 1q21 and identifies a previously unreported mutation in GJA8. These findings thus expand the mutation spectrum of GJA8. PMID- 17110921 TI - A guide for diagnostic evaluations. AB - Accurate diagnostic tests have a key role in patient management and the control of most infectious diseases. Unfortunately, in many developing countries, clinical care is often critically compromised by the lack of regulatory controls on the quality of these tests. The information available on the performance of a diagnostic test can be biased or flawed because of failings in the design of the studies which assessed the performance characteristics of the test. As a result, diagnostic tests are sold and used in much of the developing world without evidence of effectiveness. Misdiagnosis leading to failure to treat a serious infection or wasting expensive treatment on people who are not infected remains a serious obstacle to health. PMID- 17110922 TI - Why do we need quality-assured diagnostic tests for sexually transmitted infections? AB - The bacterial sexually transmitted infections (STIs) syphilis, gonorrhoea and chlamydia can all be cured with a single dose of antibiotic. Unfortunately, however, these infections often remain undiagnosed as many infected individuals have few if any symptoms. Diagnostic tests with high sensitivity and specificity are available for all three infections but, owing to their expense and the lack of laboratory capacity, most people in developing countries do not have access to these tests. There is a great need for simple, cheap diagnostic tests for STIs that can be performed at the point of care, enabling treatment to be given immediately. It is hoped that recent advances in our understanding of the pathogenesis of these infections, and the availability of the complete genome sequences for each causative organism, will lead to the development of improved point-of-care tests that will reduce the burden of these diseases in developing countries. PMID- 17110923 TI - Cytosolic factor- and TOM-independent import of C-tail-anchored mitochondrial outer membrane proteins. AB - C-tail-anchored (C-TA) proteins are anchored to specific organelle membranes by a single transmembrane segment (TMS) at the C-terminus, extruding the N-terminal functional domains into the cytoplasm in which the TMS and following basic segment function as the membrane-targeting signals. Here, we analyzed the import route of mitochondrial outer membrane (MOM) C-TA proteins, Bak, Bcl-XL, and Omp25, using digitonin-permeabilized HeLa cells, which provide specific and efficient import under competitive conditions. These experiments revealed that (i) C-TA proteins were imported to the MOM through a common pathway independent of the components of the preprotein translocase of the outer membrane, (ii) the C TA protein-targeting signal functioned autonomously in the absence of cytoplasmic factors that specifically recognize the targeting signals and deliver the preproteins to the MOM, (iii) the function of a cytoplasmic chaperone was required if the cytoplasmic domains of the C-TA proteins assumed an import incompetent conformation, and intriguingly, (iv) the MOM-targeting signal of Bak, in the context of the Bak molecule, required activation by the interaction of its cytoplasmic domain with VDAC2 before MOM targeting. PMID- 17110924 TI - AKAP150, a switch to convert mechano-, pH- and arachidonic acid-sensitive TREK K(+) channels into open leak channels. AB - TREK channels are unique among two-pore-domain K(+) channels. They are activated by polyunsaturated fatty acids (PUFAs) including arachidonic acid (AA), phospholipids, mechanical stretch and intracellular acidification. They are inhibited by neurotransmitters and hormones. TREK-1 knockout mice have impaired PUFA-mediated neuroprotection to ischemia, reduced sensitivity to volatile anesthetics and altered perception of pain. Here, we show that the A-kinase anchoring protein AKAP150 is a constituent of native TREK-1 channels. Its binding to a key regulatory domain of TREK-1 transforms low-activity outwardly rectifying currents into robust leak conductances insensitive to AA, stretch and acidification. Inhibition of the TREK-1/AKAP150 complex by Gs-coupled receptors such as serotonin 5HT4sR and noradrenaline beta2AR is as extensive as for TREK-1 alone, but is faster. Inhibition of TREK-1/AKAP150 by Gq-coupled receptors such as serotonin 5HT2bR and glutamate mGluR5 is much reduced when compared to TREK-1 alone. The association of AKAP150 with TREK channels integrates them into a postsynaptic scaffold where both G-protein-coupled membrane receptors (as demonstrated here for beta2AR) and TREK-1 dock simultaneously. PMID- 17110925 TI - Increased frequency of homologous recombination and T-DNA integration in Arabidopsis CAF-1 mutants. AB - Chromatin assembly factor 1 (CAF-1) is involved in nucleo some assembly following DNA replication and nucleotide excision repair. In Arabidopsis thaliana, the three CAF-1 subunits are encoded by FAS1, FAS2 and, most likely, MSI1, respectively. In this study, we asked whether genomic stability is altered in fas1 and fas2 mutants that are lacking CAF-1 activity. Depletion of either subunit increased the frequency of somatic homologous recombination (HR) in planta approximately 40-fold. The frequency of transferred DNA (T-DNA) integration was also elevated. A delay in loading histones onto newly replicated or repaired DNA might make these DNA stretches more accessible, both to repair enzymes and to foreign DNA. Furthermore, fas mutants exhibited increased levels of DNA double-strand breaks, a G2-phase retardation that accelerates endoreduplication, and elevated levels of mRNAs coding for proteins involved in HR-all factors that could also contribute to upregulation of HR frequency in fas mutants. PMID- 17110926 TI - Crystal structures of leucyl/phenylalanyl-tRNA-protein transferase and its complex with an aminoacyl-tRNA analog. AB - Eubacterial leucyl/phenylalanyl-tRNA protein transferase (L/F-transferase), encoded by the aat gene, conjugates leucine or phenylalanine to the N-terminal Arg or Lys residue of proteins, using Leu-tRNA(Leu) or Phe-tRNA(Phe) as a substrate. The resulting N-terminal Leu or Phe acts as a degradation signal for the ClpS-ClpAP-mediated N-end rule protein degradation pathway. Here, we present the crystal structures of Escherichia coli L/F-transferase and its complex with an aminoacyl-tRNA analog, puromycin. The C-terminal domain of L/F-transferase consists of the GCN5-related N-acetyltransferase fold, commonly observed in the acetyltransferase superfamily. The p-methoxybenzyl group of puromycin, corresponding to the side chain of Leu or Phe of Leu-tRNA(Leu) or Phe-tRNA(Phe), is accommodated in a highly hydrophobic pocket, with a shape and size suitable for hydrophobic amino-acid residues lacking a branched beta-carbon, such as leucine and phenylalanine. Structure-based mutagenesis of L/F-transferase revealed its substrate specificity. Furthermore, we present a model of the L/F transferase complex with tRNA and substrate proteins bearing an N-terminal Arg or Lys. PMID- 17110927 TI - Structural mechanism of RPA loading on DNA during activation of a simple pre replication complex. AB - We report that during activation of the simian virus 40 (SV40) pre-replication complex, SV40 T antigen (Tag) helicase actively loads replication protein A (RPA) on emerging single-stranded DNA (ssDNA). This novel loading process requires physical interaction of Tag origin DNA-binding domain (OBD) with the RPA high affinity ssDNA-binding domains (RPA70AB). Heteronuclear NMR chemical shift mapping revealed that Tag-OBD binds to RPA70AB at a site distal from the ssDNA binding sites and that RPA70AB, Tag-OBD, and an 8-nucleotide ssDNA form a stable ternary complex. Intact RPA and Tag also interact stably in the presence of an 8 mer, but Tag dissociates from the complex when RPA binds to longer oligonucleotides. Together, our results imply that an allosteric change in RPA quaternary structure completes the loading reaction. A mechanistic model is proposed in which the ternary complex is a key intermediate that directly couples origin DNA unwinding to RPA loading on emerging ssDNA. PMID- 17110928 TI - The HECT ubiquitin ligase AIP4 regulates the cell surface expression of select TRP channels. AB - TRPV4 is a widely expressed member of the transient receptor potential (TRP) family that facilitates Ca(2+) entry into nonexcitable cells. TRPV4 is activated by several stimuli, but it is largely unknown how the activity of this channel is terminated. Here, we show that ubiquitination represents an important mechanism to control the presence of TRPV4 at the plasma membrane. Ubiquitination of TRPV4 is dramatically increased by the HECT (homologous to E6-AP carboxyl terminus) family ubiquitin ligase AIP4 without inducing degradation of this channel. Instead, AIP4 promotes the endocytosis of TRPV4 and decreases its amount at the plasma membrane. Consequently, the basal activity of TRPV4 is reduced despite an overall increase in TRPV4 levels. This mode of regulation is not limited to TRPV4. TRPC4, another member of the TRP channel family, is also strongly ubiquitinated in the presence of AIP4, leading to the increased intracellular localization of TRPC4 and the reduction of its basal activity. However, ubiquitination of several other TRP channels is not affected by AIP4, demonstrating that AIP4-mediated regulation is a unique property of select TRP channels. PMID- 17110929 TI - MDMX regulation of p53 response to ribosomal stress. AB - Ribosomal stress such as disruption of rRNA biogenesis activates p53 by release of ribosomal proteins from the nucleoli, which bind to MDM2 and inhibit p53 degradation. We found that p53 activation by ribosomal stress requires degradation of MDMX in an MDM2-dependent fashion. Tumor cells overexpressing MDMX are less sensitive to actinomycin D-induced growth arrest due to formation of inactive p53-MDMX complexes. Knockdown of MDMX increases sensitivity to actinomycin D, whereas MDMX overexpression abrogates p53 activation and prevents growth arrest. Furthermore, MDMX expression promotes resistance to the chemotherapeutic agent 5-fluorouracil (5-FU), which at low concentrations activates p53 by inducing ribosomal stress without significant DNA damage signaling. Knockdown of MDMX abrogates HCT116 tumor xenograft formation in nude mice. MDMX overexpression does not accelerate tumor growth but increases resistance to 5-FU treatment in vivo. Therefore, MDMX is an important regulator of p53 response to ribosomal stress and RNA-targeting chemotherapy agents. PMID- 17110930 TI - An antiapoptotic protein, c-FLIPL, directly binds to MKK7 and inhibits the JNK pathway. AB - Inhibition of NF-kappaB activation increases susceptibility to tumor necrosis factor (TNF)alpha-induced cell death, concurrent with caspases and prolonged c Jun N-terminal kinase (JNK) activation, and reactive oxygen species (ROS) accumulation. However, the detailed mechanisms are unclear. Here we show that cellular FLICE-inhibitory protein (c-FLIP) is rapidly lost in NF-kappaB activation-deficient, but not wild-type fibroblasts upon TNFalpha stimulation, indicating that NF-kappaB normally maintains the cellular levels of c-FLIP. The ectopic expression of the long form of c-FLIP (c-FLIPL) inhibits TNFalpha-induced prolonged JNK activation and ROS accumulation in NF-kappaB activation-deficient fibroblasts. Conversely, TNFalpha induces prolonged JNK activation and ROS accumulation in c-Flip-/- fibroblasts. Moreover, c-FLIPL directly interacts with a JNK activator, MAP kinase kinase (MKK)7, in a TNFalpha-dependent manner and inhibits the interactions of MKK7 with MAP/ERK kinase kinase 1, apoptosis-signal regulating kinase 1, and TGFbeta-activated kinase 1. This stimuli-dependent interaction of c-FLIPL with MKK7 might selectively suppress the prolonged phase of JNK activation. Taken that ROS promote JNK activation and activation of the JNK pathway may promote ROS accumulation, c-FLIPL might block this positive feedback loop, thereby suppressing ROS accumulation. PMID- 17110931 TI - Allosteric activation of the protein kinase PDK1 with low molecular weight compounds. AB - Organisms rely heavily on protein phosphorylation to transduce intracellular signals. The phosphorylation of a protein often induces conformational changes, which are responsible for triggering downstream cellular events. Protein kinases are themselves frequently regulated by phosphorylation. Recently, we and others proposed the molecular mechanism by which phosphorylation at a hydrophobic motif (HM) regulates the conformation and activity of many members of the AGC group of protein kinases. Here we have developed specific, low molecular weight compounds, which target the HM/PIF-pocket and have the ability to allosterically activate phosphoinositide-dependent protein kinase 1 (PDK1) by modulating the phosphorylation-dependent conformational transition. The mechanism of action of these compounds was characterized by mutagenesis of PDK1, synthesis of compound analogs, interaction-displacement studies and isothermal titration calorimetry experiments. Our results raise the possibility of developing drugs that target the AGC kinases via a novel mode of action and may inspire future rational development of compounds with the ability to modulate phosphorylation-dependent conformational transitions in other proteins. PMID- 17110932 TI - RNA polymerase II bypass of oxidative DNA damage is regulated by transcription elongation factors. AB - Oxidative lesions represent the most abundant DNA lesions within the cell. In the present study, we investigated the impact of the oxidative lesions 8-oxoguanine, thymine glycol and 5-hydroxyuracil on RNA polymerase II (RNA pol II) transcription using a well-defined in vitro transcription system. We found that in a purified, reconstituted transcription system, these lesions block elongation by RNA pol II to different extents, depending on the type of lesion. Suggesting the presence of a bypass activity, the block to elongation is alleviated when transcription is carried out in HeLa cell nuclear extracts. By purifying this activity, we discovered that TFIIF could promote elongation through a thymine glycol lesion. The elongation factors Elongin and CSB, but not TFIIS, can also stimulate bypass of thymine glycol lesions, whereas Elongin, CSB and TFIIS can all enhance bypass of an 8-oxoguanine lesion. By increasing the efficiency with which RNA pol II reads through oxidative lesions, elongation factors can contribute to transcriptional mutagenesis, an activity that could have implications for the generation or progression of human diseases. PMID- 17110933 TI - Structural basis of actin filament capping at the barbed-end: a cryo-electron microscopy study. AB - The intracellular distribution and migration of many protein complexes and organelles is regulated by the dynamics of the actin filament. Many actin filament end-binding proteins play crucial roles in actin dynamics, since polymerization and depolymerization of actin protomers occur only at the filament ends. We present here an EM structure of the complex of the actin filament and hetero-dimeric capping protein (CP) bound to the barbed-end at 23 A resolution, by applying a newly developed methods of image analysis to cryo-electron micrographs. This structure was fitted by the crystal structure of CP and the proposed actin filament structure, allowing us to construct a model that depicts two major binding regions between CP and the barbed-end. This binding scheme accounted for the results of newly performed and previously published mutation experiments, and led us to propose a two-step binding model. This is the first determination of an actin filament end structure. PMID- 17110934 TI - Depth dependence of stratum corneum lipid ordering: a slow-tumbling simulation for electron paramagnetic resonance. AB - We investigated the structural ordering of stratum corneum (SC) lipid by means of electron paramagnetic resonance (EPR) slow-tumbling simulation in conjunction with spin probe studies. The SC of human mid-volar forearm was stripped consecutively from three to six times. The EPR probe method detected a characteristic peak of sebaceous matter in the first SC stripping. The order parameter values obtained by the slow-tumbling simulation (S(0)) showed significant differences between each layer compared with those indicated by the conventional order parameter (S) using hyperfine couplings. Although the conventional S values were in the range of 0.56 (outermost layer) to 0.61 (bottom layer), the S(0) values by the simulation changed from 0.22 to 0.96. The present results suggest that the structural ordering of the outermost SC layer is less tight, whereas the structure of inner layers becomes more rigid. Therefore, we concluded that the EPR probe method recognizes sebaceous matters, whereas EPR in conjunction with the simulation allows quantitative evaluation of SC lipid ordering in relation to skin depth. PMID- 17110935 TI - Role of aPKC isoforms and their binding partners Par3 and Par6 in epidermal barrier formation. AB - The skin water barrier, essential for terrestrial life, is formed by a multilayered stratifying epithelium, which shows a polarized distribution of both differentiation and intercellular junction markers. Recently, several reports showed the crucial importance of tight junctions for the in vivo water barrier function of the skin. In simple epithelial cells, intercellular junction formation is closely coupled to the establishment of polarity. However, if and how polarity proteins contribute to epidermal differentiation and junction formation is not yet known. Here, we have characterized the localization and isoform expression of the polarity protein atypical PKC (aPKC) and its binding partners Par3 and Par6 in epidermis and primary keratinocytes of mice. Their distribution is only partially overlapping in the granular layer, the site of functional tight junctions, suggesting that next to a common Par3/Par6/aPKC function they also may have functions independent of each other. Both aPKCzeta and aPKCiota/lambda, are expressed in the epidermis but only aPKCiota/lambda showed a strong enrichment in the junctions, suggesting that this aPKC isoform is important for epidermal tight junction function. Indeed, inhibition of aPKC function showed that endogenous aPKC is crucial for in vitro barrier function and this required the presence of both the Par3 and Par6 binding sites. PMID- 17110936 TI - Plakoglobin deficiency protects keratinocytes from apoptosis. AB - The armadillo family protein plakoglobin (Pg) is a well-characterized component of anchoring junctions, where it functions to mediate cell-cell adhesion and maintain epithelial tissue integrity. Although its closest homolog beta-catenin acts in the Wnt signaling pathway to dictate cell fate and promote proliferation and survival, the role of Pg in these processes is not well understood. Here, we investigate how Pg affects the survival of mouse keratinocytes by challenging both Pg-null cells and their heterozygote counterparts with apoptotic stimuli. Our results indicate that Pg deletion protects keratinocytes from apoptosis, with null cells exhibiting delayed mitochondrial cytochrome c release and activation of caspase-3. Pg-null keratinocytes also exhibit increased messenger RNA and protein levels of the anti-apoptotic molecule Bcl-X(L) compared to heterozygote controls. Importantly, reintroduction of Pg into the null cells shifts their phenotype towards that of the Pg+/- keratinocytes, providing further evidence that Pg plays a direct role in regulating cell survival. Taken together, our results suggest that in addition to its adhesive role in epithelia, Pg may also function in contrast to the pro-survival tendencies of beta-catenin, to potentiate death in cells damaged by apoptotic stimuli, perhaps limiting the potential for the propagation of mutations and cellular transformation. PMID- 17110937 TI - Pseudoxanthoma elasticum-like phenotype with cutis laxa and multiple coagulation factor deficiency represents a separate genetic entity. AB - Data on six patients with a Pseudoxanthoma Elasticum (PXE)-like phenotype, characterized by excessive skin folding (resembling cutis laxa) and a deficiency of the vitamin K-dependent clotting factors (II, VII, IX, and X) are presented. A comparison is made between the clinical, ultrastructural, and molecular findings in these patients and those seen in classic PXE and cutis laxa, respectively. Clinical overlap with PXE is obvious from the skin manifestations of yellowish papules or leathery plaques with dot-like depressions at presentation, angioid streaks and/or ocular peau d'orange, and fragmentation and calcification of elastic fibers in the dermis. Important phenotypic differences with PXE include much more severe skin laxity with spreading toward the trunk and limbs with thick, leathery skin folds rather than confinement to flexural areas, and no decrease in visual acuity. Moreover, detailed electron microscopic analyses revealed that alterations of elastic fibers as well as their mineralization were slightly different from those in classic PXE. Molecular analysis revealed neither causal mutations in the ABCC6 gene (ATP-binding cassette subfamily C member 6), which is responsible for PXE, nor in VKORC1 (vitamin K 2,3 epoxide reductase), known to be involved in vitamin K-dependent factor deficiency. However, the GGCX gene (gamma-glutamyl carboxylase), encoding an enzyme important for gamma carboxylation of gla-proteins, harbored mutations in six out of seven patients analyzed. These findings all support the hypothesis that the disorder indeed represents a separate clinical and genetic entity, the molecular background of which remains to be unraveled. PMID- 17110938 TI - The innate immune system 'puzzle'. AB - Detecting pathogen invasion and regulating homeostatic processes are two essential functions of myriad non-Toll-like innate immune proteins. PMID- 17110939 TI - The innate signaling of dangers and the dangers of innate signaling. AB - The innate immune system of mammals has been forged by coevolution with microbes in response to the double constraint of preserving a symbiotic interaction with commensal flora and eliminating intrusion of those commensals or invasion by pathogens. Thus, a 'sensing' network, accompanied by or lacking inflammatory responses, is controlled by elaborate mechanisms of regulation that maintain balance in the basal state. A growing number of non-Toll-like innate immune receptors is recognized as part of this surveillance network. PMID- 17110940 TI - Plant NBS-LRR proteins in pathogen sensing and host defense. AB - Plant proteins belonging to the nucleotide-binding site-leucine-rich repeat (NBS LRR) family are used for pathogen detection. Like the mammalian Nod-LRR protein 'sensors' that detect intracellular conserved pathogen-associated molecular patterns, plant NBS-LRR proteins detect pathogen-associated proteins, most often the effector molecules of pathogens responsible for virulence. Many virulence proteins are detected indirectly by plant NBS-LRR proteins from modifications the virulence proteins inflict on host target proteins. However, some NBS-LRR proteins directly bind pathogen proteins. Association with either a modified host protein or a pathogen protein leads to conformational changes in the amino terminal and LRR domains of plant NBS-LRR proteins. Such conformational alterations are thought to promote the exchange of ADP for ATP by the NBS domain, which activates 'downstream' signaling, by an unknown mechanism, leading to pathogen resistance. PMID- 17110941 TI - Nod-like proteins in immunity, inflammation and disease. AB - The intracellular Nod-like proteins or receptors are a family of sensors of intracellularly encountered microbial motifs and 'danger signals' that have emerged as being critical components of the innate immune responses and of inflammation in mammals. Several Nod-like receptors, including Nod1, Nod2, NALP3, Ipaf and Naip, are strongly associated with host responses to intracellular invasion by bacteria or the intracellular presence of specific bacterial products. An additional key function of Nod-like receptors is in inflammatory conditions, which has been emphasized by the identification of several different mutations in the genes encoding Nod1, Nod2 and NALP3 that are associated with susceptibility to inflammatory disorders. Those and other issues related to the Nod-like receptor family are discussed here. PMID- 17110942 TI - Myeloid C-type lectins in innate immunity. AB - C-type lectins expressed on myeloid cells comprise a family of proteins that share a common structural motif, and some act as receptors in pathogen recognition. But just as the presence of leucine-rich repeats alone is not sufficient to define a Toll-like receptor, the characterization of C-type lectin receptors in innate immunity requires the identification of accompanying signaling motifs. Here we focus on the known signaling pathways of myeloid C-type lectins and on their possible functions as autonomous activating or inhibitory receptors involved in innate responses to pathogens or self. PMID- 17110943 TI - The TREM receptor family and signal integration. AB - TREM proteins are a family of cell surface receptors that participate in diverse cell processes, including inflammation, bone homeostasis, neurological development and coagulation. TREM-1, the first to be identified, acts to amplify inflammation. Other TREM proteins regulate the differentiation and function of macrophages, microglia, dendritic cells, osteoclasts and platelets. Here we discuss the state of the field, putative ligands of TREM proteins and the challenges that remain in understanding TREM biology. PMID- 17110945 TI - From antigen processing to peptide-MHC binding. AB - Emil R. Unanue recounts his seminal work 25 years ago demonstrating that antigens are phagocytosed and processed by antigen-presenting cells and that class II major histocompatibility complex molecules are peptide-binding proteins. PMID- 17110946 TI - Immunological correlates of protection from HIV infection and disease. AB - The recent meeting on "Immune Correlates of Protection from HIV Infection and Disease" examined new data from a variety of preclinical and clinical settings. These new insights may facilitate vaccine design and clinical evaluation. PMID- 17110947 TI - Fetal thymocyte development: putting a zinc finger on it? PMID- 17110948 TI - Integrins and ITAMs: more than just good neighbors. PMID- 17110949 TI - Turning up the heat on HEVs. PMID- 17110953 TI - The protein kinase TOUSLED is required for maintenance of transcriptional gene silencing in Arabidopsis. AB - TOUSLED-like kinases (TLKs) are highly conserved in plants and animals, but direct evidence linking TLKs and transcriptional gene silencing is lacking. We isolated two new alleles of TOUSLED (TSL). Mutations of TSL in ros1 reactivate the transcriptionally silent 35S-NPTII transgene and the transcriptionally silent endogenous loci TSI (TRANSCRIPTIONAL SILENCING INFORMATION). Chromatin immunoprecipitation (ChIP) analysis shows that histone H3Lys9 dimethylation is decreased in the reactivated transgene and endogenous TSI loci in the tsl ros1 mutant. However, there is no change in DNA methylation in the affected loci. Western blot and ChIP assay suggest that TSL might not be responsible for histone H3Ser10 phosphorylation. The tsl seedlings were more sensitive to DNA damage reagent methyl methanesulphonate and UV-B light. Our results provide direct evidence for a crucial role of the TOUSLED protein kinase in the maintenance of transcriptional gene silencing in some genomic regions in a DNA-methylation independent manner in Arabidopsis. PMID- 17110954 TI - Nuclear export modulates the cytoplasmic Sir2 homologue Hst2. AB - Modulating transcription factors is crucial to executing sophisticated gene expression programs. The silent information regulator 2 (Sir2) family of NAD dependent protein deacetylases influences transcription by targeting proteins such as histones, p53 and forkhead-box family transcription factors. Although apparently cytoplasmic, both mammalian SIRT2 and its yeast orthologue Hst2 have been implicated in transcriptional regulation. Here, we show that Hst2 moves between the nucleus and cytoplasm, but is largely cytoplasmic owing to efficient nuclear export. This nuclear exclusion is mediated by the exportin chromosomal region maintenance 1 (Crm1) and a putative leucine-rich nuclear export sequence in Hst2, which overlaps a unique autoregulatory helix. Disruption of Hst2 export shows that nuclear exclusion inhibits the activity of Hst2 as a transcriptional repressor. Our identification of putative nuclear export sequences in numerous vertebrate SIRT2 proteins shows that active nuclear export can be a conserved mechanism for regulating Sir2 homologues. PMID- 17110955 TI - Protozoan encounters with Toll-like receptor signalling pathways: implications for host parasitism. AB - Toll-like receptors (TLRs) have emerged as a major receptor family involved in non-self recognition. They have a vital role in triggering innate immunity and orchestrate the acquired immune response during bacterial and viral infection. However, the role of TLRs during infection with protozoan pathogens is less clear. Nevertheless, our understanding of how these parasitic microorganisms engage the host TLR signalling system has now entered a phase of rapid expansion. This Review describes recent insights into how parasitic protozoans are sensed by TLR molecules, and how the TLR system itself can be targeted by these microbial pathogens for their own survival. PMID- 17110956 TI - Structural determinants of T-cell receptor bias in immunity. AB - Antigen-specific T-cell responses induced by infection, transplantation, autoimmunity or hypersensitivity are characterized by cells expressing biased profiles of T-cell receptors (TCRs) that are selected from a diverse, naive repertoire. Here, we review the evidence for these TCR biases, focusing on crystallographic analysis of the structural constraints that determine the binding of a TCR to its ligand and the persistence of certain TCRs in an immune repertoire. We discuss the ways in which diversity in a selected TCR repertoire can contribute to protective immunity and the implications of this for vaccine design and immunotherapy. PMID- 17110957 TI - Inhibition of urokinase-type plasminogen activator receptor induces apoptosis in melanoma cells by activation of p53. AB - The urokinase-type plasminogen activator receptor (uPAR) is involved in several biological processes, including proteolysis, adhesion, migration and inflammation. Increased expression of uPAR is associated with metastasis in several tumor types. We studied the biological role of uPAR in melanoma and found that inhibition of uPAR via RNA interference induced massive death in three different metastatic cell lines. Annexin-V staining and caspase activation analysis revealed induction of the mitochondrial apoptotic pathway. The expression of members of the Bcl-2 family (Bax, Bcl-2, Bak and Bcl-x(L)) was changed in a pro-apoptotic manner. uPAR inhibition induced the expression of the tumor suppressor p53 and of its downstream target gene p21. Inhibition of p53 rescued cells from apoptosis indicating that p53 was critical for apoptosis induction. Apoptosis was observed in melanoma cells carrying activating BRAF mutations and occurred in the presence of extracellular signal-regulated kinase (ERK) phosphorylation. uPAR can activate focal adhesion kinase (FAK), which is implicated in adhesion-dependent tumor cell survival. However, inhibition of FAK did not induce apoptosis. Our data suggest a new function of uPAR acting as a survival factor for melanoma by downregulating p53. Inhibition of uPAR induces a pro-apoptotic signalling pathway via p53 that is independent of ERK or FAK signalling. These findings may offer new treatment strategies for metastatic melanoma. PMID- 17110958 TI - The Yes-associated protein 1 stabilizes p73 by preventing Itch-mediated ubiquitination of p73. AB - Upon DNA damage signaling, p73, a member of the p53 tumor suppressor family, accumulates to support transcription of downstream apoptotic genes. p73 interacts with Yes-associated protein 1 (Yap1) through its PPPY motif, and increases p73 transactivation of apoptotic genes. The ubiquitin E3 ligase Itch, like Yap1, interacts with p73. Given the fact that both Itch and Yap1 bind p73 via the PPPY motif, we hypothesized that Yap may also function to stabilize p73 by displacing Itch binding to p73. We show that the interaction of Yap1 and p73 was necessary for p73 stabilization. Yap1 competed with Itch for binding to p73, and prevented Itch-mediated ubiquitination of p73. Treatment of cells with cisplatin leads to an increase in p73 accumulation and induction of apoptosis, but both were dramatically reduced in the presence of Yap1 siRNA. Altogether, our findings attribute a central role to Yap1 in regulating p73 accumulation and function under DNA damage signaling. PMID- 17110959 TI - Effect of acidic electrolyzed water on the viability of bacterial and fungal plant pathogens and on bacterial spot disease of tomato. AB - Acidic electrolyzed water (AEW), known to have germicidal activity, was obtained after electrolysis of 0.045% aqueous solution of sodium chloride. Freshly prepared AEW (pH 2.3-2.6, oxidation-reduction potential 1007-1025 mV, and free active chlorine concentration 27-35 ppm) was tested in vitro and (or) on tomato foliage and seed surfaces for its effects on the viability of plant pathogen propagules that could be potential seed contaminants. Foliar sprays of AEW were tested against bacterial spot disease of tomato under greenhouse and field conditions. The viability of propagules of Xanthomonas campestris pv. vesicatoria (bacterial spot pathogen), Streptomyces scabies (potato scab pathogen), and Fusarium oxysporum f.sp. lycopersici (root rot pathogen) was significantly reduced 4-8 log units within 2 min of exposure to AEW. Immersion of tomato seed from infected fruit in AEW for 1 and 3 min significantly reduced the populations of X. campestris pv. vesicatoria from the surface of the seed without affecting seed germination. Foliar sprays of AEW reduced X. campestris pv. vesicatoria populations and leaf spot severity on tomato foliage in the greenhouse. In the field, multiple sprays of AEW consistently reduced bacterial spot severity on tomato foliage. Disease incidence and severity was also reduced on fruit, but only in 2003. Fruit yield was either enhanced or not affected by the AEW sprays. These results indicate a potential use of AEW as a seed surface disinfectant or contact bactericide. PMID- 17110960 TI - Dietary fructooligosaccharides alter the cultivable faecal population of rats but do not stimulate the growth of intestinal bifidobacteria. AB - The effect of fructans on the cultivable faecal community of Bio Breeding rats fed diets containing 5% (m/v) food-grade fructooligosaccharide (FOS) was investigated. Culturing of faecal material using chicory inulin as the sole carbohydrate source revealed the presence of a greater diversity of inulin utilizing bacterial species in FOS-fed rats as compared with the control rats, although both contained species which effectively utilized inulin. The majority of cultivable inulin-utilizing species fell within the Clostridium coccoides group and Clostridium leptum subgroup, some of which were related to previously cultured butyrate-producing bacteria from the intestines of various animals. The impact of FOS on the growth of the indigenous bifidobacteria community and three inulin-utilizing isolates was assessed using real-time polymerase chain reaction. While dietary FOS was found to stimulate the growth of all three inulin-utilizing isolates, no growth stimulation of the indigenous bifidobacteria community occurred over the duration of the feeding trial. PMID- 17110961 TI - Identification and characterization of a polysaccharide deacetylase gene from Bacillus thuringiensis. AB - One polysaccharide deacetylase gene was cloned from Bacillus thuringiensis and designated pdaA. Disruption of pdaA did not affect vegetative growth and sporulation but obviously affected spore germination. When L-alanine was added into the spore suspension, the spores of the pdaA disruption mutant showed a slow and partial reduction in absorbance at OD600 and became phase pale gray compared with phase dark of the wild-type strain. In contrast with the outgrowing of wild type spores after germination, the pdaA mutant spores were blocked at the stage of spore germination. Transmission electron micrographs revealed a significant difference between the pdaA mutant and the wild-type strain in the spore cortex. Introduction of the pdaA gene into the pdaA disruption mutant complemented the germination-negative phenotype. Reverse transcription--polymerase chain reaction showed that pdaA was transcribed after incubation for 10 h in CCY medium. PMID- 17110962 TI - Occurrence of ceftriaxone-resistant commensal bacteria on a dairy farm and a poultry farm. AB - Approximately 40 samples of animal feces, drinking water, feed, bedding, pine wood shavings, compost, and manure slurry were collected from two animal research farms (one dairy and one poultry) and analyzed for ceftriaxone-resistant bacteria. Our study revealed that the total percentage of aerobic bacteria with reduced susceptibility to ceftriaxone (minimal inhibitory concentration (MIC) > or = 16 micro g/mL) ranged from 0.9% to 10.8% in dairy feces and from 0.05% to 3.93% in chicken feces. The percentages of ceftriaxone-resistant bacteria (MIC > or = 64 micro g/mL) were in the range of 0.01% - 2.3% in dairy feces and 0.01% - 0.79% in chicken feces. Environmental samples contained a wide range of ceftriaxone-resistant bacterial populations. Among those environmental samples, fresh pine wood shavings used as chicken bedding contained the highest percentages (41.5%) of ceftriaxone-resistant bacteria, as determined by a plating method. A total of 105 ceftriaxone-resistant (MIC > or = 128 micro g/mL) bacterial isolates were isolated from the above samples and tested for resistance to nine antibiotics: ampicillin, ceftriaxone, streptomycin, kanamycin, gentamicin, chloramphenicol, tetracycline, ciprofloxacin, and nalidixic acid. The most prevalent resistance pattern (34.3%) among isolates included resistance to all nine antibiotics. Results from this study suggest that ceftriaxone-resistant bacteria exist in farm environments, and the ceftriaxone resistance was frequently associated with resistance to multiple antibiotics. Environmental sources such as pine wood shavings used as bedding can be a potential reservoir for transmitting the multidrug-resistant bacteria. PMID- 17110963 TI - Relationship between soil densities of Aspergillus species and colonization of wounded peanut seeds. AB - Soil is a reservoir for Aspergillus flavus and A. parasiticus, fungi that commonly colonize peanut seeds and produce carcinogenic aflatoxins. Densities of these fungi in soil vary greatly among fields and may influence the severity of peanut infection. This study examined the relationship between soil density of Aspergillus species and the incidence of peanut seed colonization under laboratory conditions. Viable peanut seeds were wounded and inoculated with 20 soils differing in composition and density of Aspergillus species and were then incubated for 14 days at 37 degrees C (seed water activity = 0.92). The effect of soil density of individual section Flavi species (A. flavus strains L and S, A. parasiticus, A. caelatus, and A. tamarii), section Nigri, and A. terreus on the incidence of seed colonization was best expressed as a function of exponential rise to maximum. Exponential curves often rose to maximum percentages of seed colonization by section Flavi species that were well below 100% despite high species densities in some soils. Competition primarily among section Flavi species may explain the reduced incidences of seed colonization. An average of two or fewer propagules of each Aspergillus species in the soil at the wound site was required for colonization of 20% of peanut seeds. Other fungal species were capable of invading peanut seeds only when soil densities of sections Flavi and Nigri species were low. PMID- 17110964 TI - Chitinase production during interaction of Trichoderma aggressivum and Agaricus bisporus. AB - The competitor fungus Trichoderma aggressivum causes green mould disease, a potentially devastating problem of the commercial mushroom Agaricus bisporus. Due to the recent appearance of this problem, very little is known about the mechanisms by which T. aggressivum interacts with and inhibits A. bisporus. A mechanism generally used by Trichoderma species in the antagonism of other fungi is the secretion of cell wall degrading enzymes. In this study, we determined the activities of chitinases produced in dual cultures of these fungi over a 2 week period. Both intracellular and extracellular enzymes were studied. Agaricus bisporus produced N-acetylglucosaminidases with apparent molecular masses of 111, 105, and 96 kDa. Two resistant brown strains produced greater activities of the 96 kDa N-acetylglucosaminidase than susceptible off-white and white strains. This result suggested that this enzyme might have a role in the resistance of commercial brown strains to green mould disease. Trichoderma aggressivum produced three N-acetylglucosaminidases with apparent molecular masses of 131, 125, and 122 kDa, a 40 kDa chitobiosidase, and a 36 kDa endochitinase. The 122 kDa N acetylglucosaminidase showed the greatest activity and may be an important predictor of antifungal activity. PMID- 17110965 TI - Genotypic characteristics of the rrn operon and genome of indigenous soybean Bradyrhizobia in cropping zones of China. AB - Four genetic assays, 16S rRNA restriction fragment length polymorphism (RFLP), 16S rRNA sequencing, 16S-23S rRNA intergenetic spacer (IGS) RFLP, and amplified fragment length polymorphism (AFLP), were conducted to determine the genotypic characteristics of 44 indigenous strains of Bradyrhizobium from soybean (Glycine max L.) cropping zones of China. The results generated from different assays showed that soybean bradyrhizobial isolates comprised four genomic groups. Group I was composed of strains mainly isolated from the North and Northeast plains of China. All four assays confirmed this group as phylogenetically divergent from all the reference strains. Strains of the group may represent a new species. Strains in Group II isolated from a variety of geographic regions were ascribed to B. liaoningense. Strains in Group III, mainly isolated from Central and East China, were closely related to the reference strains of B. japonicum. Strains in Group IV belonged to B. elkanii. PMID- 17110966 TI - Triclosan inhibition of membrane enzymes and glycolysis of Streptococcus mutans in suspensions and biofilms. AB - Triclosan was found to be a potent inhibitor of the F(H+)-ATPase of the oral pathogen Streptococcus mutans and to increase proton permeabilities of intact cells. Moreover, it acted additively with weak-acid transmembrane proton carriers, such as fluoride or sorbate, to sensitize glycolysis to acid inhibition. Even at neutral pH, triclosan could inhibit glycolysis more directly as an irreversible inhibitor of the glycolytic enzymes pyruvate kinase, lactic dehydro genase, aldolase, and the phosphoenolpyruvate:sugar phosphotransferase system (PTS). Cell glycolysis in suspensions or biofilms was inhibited in a pH dependent manner by triclosan at a concentration of about 0.1 mmol/L at pH 7, approximately the lethal concentration for S. mutans cells in suspensions. Cells in intact biofilms were almost as sensitive to triclosan inhibition of glycolysis as were cells in suspensions but were more resistant to killing. Targets for irreversible inhibition of glycolysis included the PTS and cytoplasmic enzymes, specifically pyruvate kinase, lactic dehydrogenase, and to a lesser extent, aldolase. General conclusions are that triclosan is a multi-target inhibitor for mutans streptococci, which lack a triclosan-sensitive FabI enoyl-ACP reductase, and that inhibition of glycolysis in dental plaque biofilms, in which triclosan is retained after initial or repeated exposure, would reduce cariogenicity. PMID- 17110967 TI - [A study to assess the microbial contamination of Mya arenaria clams from the north shore of the St Lawrence River estuary, (Quebec, Canada)]. AB - The aims of the present study were to assess the microbial quality of Mya arenaria clams from the north shore of the St. Lawrence River estuary and to validate various microbial indicator microorganisms of bivalve mollusks contamination. Clams were collected from nine sites, including four harvesting sites closed by virtue of the Canadian Shellfish Sanitation Program (CSSP). Six contamination indicators (fecal coliforms, somatic coliphages, F-specific coliphages, fecal streptococci, Clostridium perfringens, and Escherichia coli) and four pathogens (Campylobacter sp., Cryptosporidium parvum, Giardia sp., and Salmonella sp.) were identified in the clams. Indicators sensibility, specificity and predictive values with respect to the presence of pathogens were calculated. Pathogenic microorganisms detection frequency in clams was important (92%). Globally, pathogens tend to be less frequently detected in opened harvesting sites (p = 0.086). Although the assessed indicators were not perfect, when F specific coliphages are associated with E. coli or fecal coliforms, a good sensibility (62%-64%) and good positive predictive value (88%) with respect to the investigated pathogens are obtained. PMID- 17110968 TI - A rapid (one day), sensitive real-time polymerase chain reaction assay for detecting Escherichia coli O157:H7 in ground beef. AB - The purpose of this study was to apply our rapid, integrated double enrichment 5' nuclease real-time polymerase chain reaction assay for the detection of Escherichia coli O157:H7 and evaluate its efficacy. The assay targeted ground beef, an important vehicle in disease epidemiology. The assay reliably determined in 8 h the presence of E. coli O157:H7 in ground beef at the level of 1 colony forming unit (CFU)/g. The sensitivity and specificity of the assay were compared with that of standard enrichment diagnostic techniques. A correlation of 100% in detection was achieved to the limit of 1 CFU/g. This assay can be used as a rapid, automatic process for identification of E. coli O157:H7 in ground beef or can be integrated with standard culture procedures, resulting in considerable cost and time savings. PMID- 17110969 TI - Hepatitis C virus infection among First Nation and non-First Nation people in Manitoba, Canada: a public health laboratory study. AB - Demographic information and laboratory test results on 136 169 clinical serum specimens submitted to the public health laboratory in Manitoba, Canada, for hepatitis C virus (HCV) testing between January 1995 and December 2003 were analyzed. The difference in the clearance rates of HCV infection, without therapeutic intervention, and the HCV genotypes infecting First Nation and non First Nation people were studied. The rates of co-infection of HCV-positive individuals with other hepatitis viruses were also compared between the two study groups. The results of the analyses of the data indicated that there was a 4.4 fold increase in the number of specimens tested and a 4.9-fold decrease in HCV antibody (anti-HCV) positive cases during the study period. The proportion of specimens submitted for testing from First Nation individuals was lower than their proportion in the Manitoba population. Our study also indicated that there was a significantly higher proportion of First Nation patients who had self limiting infection (patients cleared the infection and became HCV RNA negative without anti-HCV treatment) in comparison to non-First Nation patients. The proportion of First Nation females who had self-limiting infection was significantly higher than non-First Nation females. HCV genotype 1 infection represented more than 60% of HCV infection in Manitoba. The rate of individuals positive for the hepatitis A virus antibody in the HCV-positive population was higher among First Nation than non-First Nation individuals. On the other hand, there were more HCV-infected First Nation patients than non-First Nation patients who were not immune to the hepatitis B virus. The data indicate that fewer First Nation patients seek anti-HCV therapy in comparison to non-First Nation. In conclusion, the differences in the rates of HCV self-limiting infection between First Nation and non-First Nation individuals in Manitoba may reflect the genetic differences between the two cohorts, which may consequently affect the immune response to the HCV infection. PMID- 17110970 TI - Detection and characterization of bacteria from the potato rhizosphere degrading N-acyl-homoserine lactone. AB - Quorum sensing plays a role in the regulation of soft rot diseases caused by the plant pathogenic bacterium Pectobacterium carotovorum subsp. carotovorum. The signal molecules involved in quorum sensing in P. carotovorum subsp. carotovorum belong to the group of N-acyl homoserine lactones (AHLs). In our study, we screened bacteria isolated from the potato rhizosphere for the ability to degrade AHLs produced by P. carotovorum subsp. carotovorum. Six isolates able to degrade AHLs were selected for further studies. According to 16S rDNA sequence analysis and fatty acid methyl ester profiling, the isolates belonged to the genera Ochrobactrum, Rhodococcus, Pseudomonas, Bacillus, and Delftia. For the genera Ochrobactrum and Delftia, for the first time AHL-degrading isolates were found. Data presented in this study revealed for the first time that Ochrobactrum sp. strain A44 showed the capacity to inactivate various synthetic AHL molecules; the substituted AHLs were inactivated with a lower efficiency than the unsubstituted AHLs. Compared with the other isolates, A44 was very effective in the degradation of AHLs produced by P. carotovorum subsp. carotovorum. It was verified by polymerase chain reaction, DNA-DNA hybridization, and a lactone ring reconstruction assay that Ochrobactrum sp. strain A44 did not possess AHL lactonase activity. AHL degradation in Ochrobactrum sp. strain A44 occurred intracellularly; it was not found in the culture supernatant. AHL-degrading activity of A44 was thermo sensitive. Experiments in planta revealed that Ochrobactrum sp. strain A44 significantly inhibited the maceration of potato tuber tissue. Since A44 did not produce antibiotics, the attenuation of the decay might be due to the quenching of quorum- sensing-regulated production of pectinolytic enzymes. The strain can potentially serve to control P. carotovorum subsp. carotovorum in potato. PMID- 17110971 TI - Biocontrol of wood-rotting fungi with Streptomyces violaceusniger XL-2. AB - During the previous decade, chitinases have received increased attention because of their wide range of applications. Chito-oligomers produced by enzymatic hydrolysis of chitin have been of interest in recent years because of their broad applications in medical, agricultural, and industrial applications, such as antibacterial, antifungal, hypo cholesterolemic, and antihypertensive activity, and as food quality enhancer. Fungal cell walls being rich in chitin also enable the use of chitinases in biocontrol of fungal pathogens, as bio-fungicides. An actinomycete was isolated from the bark of trees of Dehradun in India and was later identified as Streptomyces violaceusniger. This strain exhibits strong antagonism towards various wood-rotting fungi, such as Phanerochaete chrysosporium, Postia placenta, Coriolus versicolor, and Gloeophyllum trabeum. Further, studies showed an extracellular bioactive compound was responsible for the antagonism. The conditions for the production of this biocontrol agent were optimized, and the effects of various stress factors (like nitrogen-deficient media, carbon-deficient media, etc.) were studied. The presence of chitin in the growth media was found to be an essential factor for the active production of the biocontrol agent. The pH and temperature optima for the biocontrol agent were determined. Purification and characterization of this specific biocontrol agent was performed through anion exchange chromatography using a DEAE-cellulose column, and a single protein band was obtained on a 10% sodium dodecyl sulfate polyacrylamide gel. The protein was later identified as a 28 kDa endo chitinase by MALDI-TOF (matrix-assisted laser desorption ionization-time of flight) and by a chitobiose activity assay. PMID- 17110972 TI - Hexavalent chromium removal in vitro and from industrial wastes, using chromate resistant strains of filamentous fungi indigenous to contaminated wastes. AB - Two chromate-resistant filamentous fungi, strains H13 and Ed8, were selected from seven independent fungal isolates indigenous to Cr(VI)-contaminated soil because of their ability to decrease hexavalent chromium levels in the growth medium. Morphophysiological studies identified strain H13 as a Penicillium sp. isolate and Ed8 as an Aspergillus sp. isolate. When incubated in minimal medium with glucose as a carbon source and in the presence of 50 microg/mL Cr(VI), these strains caused complete disappearance of Cr(VI) in the growth medium after about 72 h of incubation. Total chromium concentration in growth medium was constant during culture growth, and no accumulation of chromium in fungal biomass was observed. Quantitative determinations of oxidized and reduced chromium species during the reduction process revealed stoichiometric conversion of Cr(VI) to Cr(III). A decrease in Cr(VI) levels from industrial wastes was also induced by Ed8 or H13 biomass. These results indicate that chromate-resistant filamentous fungi with Cr(VI)-reducing capability could be useful for the removal of Cr(VI) contamination. PMID- 17110973 TI - Characterization of the glyoxalase I gene from the vascular wilt fungus Verticillium dahliae. AB - A glyoxalase I gene homologue (VdGLO1) was identified in the vascular wilt fungus Verticillium dahliae by sequence tag analysis of genes expressed during resting structure development. The results of the current study show that the gene encodes a putative 345 amino acid protein with high similarity to glyoxalase I, which produces S-D-lactoylglutathione from the toxic metabolic by-product methylglyoxal (MG). Disruption of the V. dahliae gene by Agrobacterium tumefaciens-mediated transformation resulted in enhanced sensitivity to MG. Mycelial growth of disruption mutants was severely reduced in the presence of 5 mmol/L MG. In contrast, spore production in liquid medium was abolished at 1 mmol/L MG, although not at physiologically relevant concentrations of 500 CFU of Rif+ bacteria (g shoot fresh mass)-1) in weeks 2 and 3, declining to <200 CFU in week 5. Again, no Rif+ bacteria could be detected in roots. No Rif+ bacteria were recovered after 14 weeks for one of the strains. It appears that the main route of bacterial entry into seedlings was through stomata and that bacteria remained in the aerial parts of plants rather than migrating to the roots. PMID- 17110978 TI - Physical and genetic map of the Spiroplasma kunkelii CR2-3x chromosome. AB - Spiroplasma kunkelii (class Mollicutes) is the characteristically helical, wall less bacterium that causes corn stunt disease. A combination of restriction enzyme analysis, pulsed-field gel electrophoresis (PFGE), and Southern hybridization analysis was used to construct a physical and genetic map of the S. kunkelii CR2-3x chromosome. The order of restriction fragments on the map was determined by analyses of reciprocal endonuclease double digests employing I CeuI, AscI, ApaI, EagI, SmaI, BssHII, BglI, and SalI; adjacent fragments were identified on two-dimensional pulsed-field electrophoresis gels. The size of the chromosome was estimated at 1550 kb. Oligonucleotide pairs were designed to prime the amplification of 26 S. kunkelii gene sequences in the polymerase chain reaction (PCR). Using PCR amplicons as probes, the locations of 27 S. kunkelii putative single-copy genes were positioned on the map by Southern hybridization analyses of chromosomal fragments separated in PFGE. The nucleotide sequence of the single ribosomal RNA operon was determined and its location mapped to a chromosomal segment bearing recognition sites for SalI, SmaI, EagI, and I-CeuI. PMID- 17110979 TI - Characterization of the arginine deiminase of Streptococcus equi subsp. zooepidemicus. AB - Streptococcus equi subsp. zooepidemicus is an important cause of infectious diseases in horses and rarely humans. Little is known about the virulence factors or protective antigens of S. equi subsp. zooepidemicus. In the present study, I designed original primers based on an alignment of the gene sagp(arcA) from Streptococcus pyogenes encoding streptococcal acid glycoprotein-arginine deiminase (SAGP/AD) to amplify the S. equi subsp. zooepidemicus counterpart sequence by polymerase chain reaction, and I analyzed the sagp(arcA) gene of the organism. Using chromosomal walking steps, I identified a contiguous eight-gene locus involved in SAGP/AD production. Their open reading frames were found to share significant homologies and to correspond closely in molecular mass to previously sequenced arc genes of S. pyogenes, thus they were designated ahrC.2 (arginine repressor), arcR (CRP/FNR transcription regulator), sagp(arcA) (streptococcal acid glycoprotein-arginine deiminase), putative acetyltransferase gene, arcB (ornithine carbamyl transferase), arcD (arginine-ornithine antiporter), arcT (Xaa-His peptidase), and arcC (carbamate kinase). The SAGP homologue of S. equi subsp. zooepidemicus (SzSAGP), encoded by arcA gene of the bacteria (arcA(SZ)), was successfully expressed in Escherichia coli and purified to homogeneity. When in vitro growth inhibitory activity of the recombinant SzSAGP was tested against MOLT-3 cells, it inhibited the growth of the cells during the 3 days of culture in a dose-dependent manner, accompanied by the induction of apoptotic cell death. The recombinant protein also possessed AD activity. By immunoblot analysis using both anti-SzSAGP-SfbI(H8) and anti SfbI(H8) sera, I was able to demonstrate that the SzSAGP protein is expressed on the streptococcal surface. PMID- 17110980 TI - Probiotic bacteria as potential detoxification tools: assessing their heavy metal binding isotherms. AB - Dietary exposure to heavy metals may have detrimental effects on human and animal health, even at low concentrations. Specific probiotic bacteria may have properties that enable them to bind toxins from food and water. We assessed the interaction of probiotic bacteria with cadmium and lead in vitro as an initial screening step to identify strains for heavy metal decontamination in food and intestinal models. Binding isotherms for cadmium and lead were characterized for Lactobacillus rhamnosus LC-705, Propionibacterium freudenreichii subsp. shermanii JS and a mix of them used by the food industry. Differences among the strains and their combinations in binding performance at a range of concentrations between 0.1 and 100 mg.L-1 were evaluated with the Langmuir model for biosorption. The effects of pH, contact time, and viability on the binding capacities were also investigated. All strains and their combinations were found to bind cadmium and lead efficiently at low concentration ranges commonly observed in foods. However, the two strains and their combinations differed significantly in their maximum binding capacities and affinities represented by the Langmuir constants Qmax and b, respectively. The binding seemed to occur instantaneously and in a pH dependent manner, which can be perfectly described by a segmented linear-plateau model. PMID- 17110981 TI - Exploration of members of Aspergillus sections Nigri, Flavi, and Terrei for feruloyl esterase production. AB - The ability of members of Aspergillus sections Nigri, Flavi, and Terrei to produce feruloyl esterases was studied according to their substrate specificity against synthetic methyl esters of hydroxycinnamic acids. Type A feruloyl esterases (FAEA), induced during growth on cereal-derived products, show a preference for the phenolic moiety of substrates that contain methoxy substitutions, as found in methyl sinapinate, whereas type B feruloyl esterases (FAEB) show a preference for the phenolic moiety of substrates that contain hydroxyl substitutions, as occurs in methyl caffeate. All the strains of Aspergillus section Nigri (e.g., A. niger and A. foetidus) were able to produce feruloyl esterases with activity profiles similar to those reported for FAEA and FAEB of A. niger when grown on oat-spelt xylan and sugar beet pulp, respectively. The two genes encoding these proteins, faeA and faeB, were identified by Southern blot analysis. The strains of Aspergillus sections Flavi (e.g., A. flavus, A. flavo-furcatus, and A. tamarii) and Terrei (e.g., A. terreus) were able to produce type A and type B enzymes. faeA was revealed in genomic DNA of these strains, and FAEA was determined by immunodetection in cultures grown in oat spelt xylan. In addition, type B enzymes, not related to faeB, were efficiently induced by oat-spelt xylan and exhibited very original activity profiles on sugar beet pulp. This work confirms that the members of the genus Aspergillus are good feruloyl esterase producers. PMID- 17110982 TI - Effect of process parameters on succinic acid production in Escherichia coli W3110 and enzymes involved in the reductive tricarboxylic acid cycle. AB - The effect of process optimization on succinic acid production by Escherichia coli W3110 and on enzymes involved in the reverse tricarboxylic acid cycle was studied. Approximately, 7.02 g L-1 of succinic acid was produced in 60 h at pH 7.0 in 500 mL anaerobic bottles containing 300 mL of the medium, wherein the sucrose concentration was 2.5%, the ratio of tryptone to ammonium hydrogen phosphate was 1:1, and the concentration of magnesium carbon ate was 1.5%. When these optimized fermentation conditions were employed in a 10 L bioreactor, 11.2 g L-1 of succinic acid was produced in 48 h. This is a 10-fold increase in succinic acid production from the initial titer of 0.94 g L-1. This clearly indicates the importance of process optimization, where by manipulating the media composition and production conditions, a remarkable increase in the production of the desired biomolecule can be obtained. The production of succinic acid is a multi-step reaction through the reverse tricarboxylic acid cycle. A linear relationship was observed between succinic acid production and the enzyme activities. The enzyme activities were found to increase in the order phospho enol-pyruvate carboxylase95% of bootstrap replications in parsimony, distance, likelihood, and similarly high posterior probabilities in bayesian analyses, one by 85%-95% according to the method of analysis, and one by 99% by one method and a majority of the other two. The endemic Pacific species, walleye pollock (Theragra chalcogramma), is more closely related to the endemic Atlantic species, Atlantic cod (Gadus macrocephalus), than either is to a second Pacific endemic, Pacific cod (Gadus macrocephalus). The walleye pollock should thus be referred to the genus Gadus as originally described (Gadus chalcogrammus Pallas 1811). Arcto-Atlantic Greenland cod, previously regarded as a distinct species (G. ogac), are a genomically distinguishable subspecies within pan-Pacific G. macrocephalus. Of the 2 endemic Arctic Ocean genera, Polar cod (Boreogadus) as the outgroup to Arctic cod (Arctogadus) and Gadus sensu lato is more strongly supported than a pairing of Boreogadus and Arctogadus as sister taxa. Taking into consideration historical patterns of hydrogeography, we outline a hypothesis of the origin of the 2 endemic Pacific species as independent but simultaneous invasions through the Bering Strait from an Arcto-Atlantic ancestral lineage. In contrast to the genome data, the complete proteome sequence (3830 amino acids) resolved only 3 nodes with >95% confidence, and placed Alaska pollock outside the Gadus clade owing to reversal mutations in the ND5 locus that restore ancestral, non-Gadus, amino acid residues in that species. PMID- 17110993 TI - Single-nucleotide polymorphism frequency in a set of selected lines of bread wheat (Triticum aestivum L.). AB - Information on single-nucleotide polymorphisms (SNPs) in hexaploid bread wheat is still scarce. The goal of this study was to detect SNPs in wheat and examine their frequency. Twenty-six bread wheat lines from different origins worldwide were used. Specific PCR-products were obtained from 21 genes and directly sequenced. SNPs were discovered from the alignment of these sequences. The overall sequence polymorphism observed in this sample appears to be low; 64 single-base polymorphisms were detected in approximately 21.5 kb (i.e., 1 SNP every 335 bp). The level of polymorphism is highly variable among the different genes studied. Fifty percent of the genes studied contained no sequence polymorphism, whereas most SNPs detected were located in only 2 genes. As expected, taking into account a synthetic line created with a wild Triticum tauschii parent increases the level of polymorphism (101 SNPs; 1 SNP every 212 bp). The detected SNPs are available at http://urgi.versailles.inra.fr/GnpSNP">http://urgi.versailles.inra.fr/GnpSNP. Data on linkage disequilibrium (LD) are still preliminary. They showed a significant level of LD in the 2 most polymorphic genes. To conclude, the genome size of hexaploid wheat and its low level of polymorphism complicate SNP discovery in this species. PMID- 17110994 TI - The Sbm1 locus conferring resistance to Soil-borne cereal mosaic virus maps to a gene-rich region on 5DL in wheat. AB - A mosaic disease caused by Soil-borne cereal mosaic virus (SBCMV) is becoming increasingly important, particularly in winter wheat in Europe. As there are currently no effective cultural practices or practical environmentally friendly chemicals for disease control, host plant resistance is an important objective in breeding programs. However, development of resistant cultivars is slow owing to difficulties in germplasm screening for resistance. Therefore, there is a need to identify molecular markers linked to SBCMV-resistance gene(s), so that quick and accurate laboratory-based marker-assisted selection rather than prolonged field based screens for resistance can be used in developing resistant cultivars. We previously demonstrated that resistance to SBCMV in Triticum aestivum 'Cadenza' is controlled by a single locus. In this work, we used AFLP and microsatellite technology to map this resistance locus, with the proposed name Sbm1, to the distal end of chromosome 5DL. Interestingly, several expressed disease-resistance gene analogues also map to this gene-rich region on 5DL. Closely linked (approximately 17 cM interval) markers, BARC110 and WMC765, RRES01 and BARC144, that flank Sbm1 will be very useful in breeding for selection of germplasm carrying Sbm1. PMID- 17110995 TI - Retrotransposon-based S-SAP as a platform for the analysis of genetic variation and linkage in globe artichoke. AB - A high copy number of retrotransposon sequences are present and widely dispersed in plant genomes. Their activity generates a considerable degree of sequence polymorphism. Here, we report the cloning of CYRE-5, a long-terminal repeat carrying retrotransposon-like sequence in Cynara cardunculus L., and its exploitation to develop a DNA fingerprinting assay across 22 accessions, including both cultivated (globe artichoke and cultivated cardoon) and wild (wild cardoon) types. The effectiveness of the sequence-specific amplified polymorphism (S-SAP) platform is compared with that of amplified fragment length polymorphism (AFLP). A genetic linkage analysis, based on a hybrid population between 2 globe artichoke varietal types, resulted in the inclusion of 29 S-SAP loci in the core genetic map, confirming their dispersed distribution across the globe artichoke genome. PMID- 17110996 TI - Molecular diversity and relationships among Elymus trachycaulus, E. subsecundus, E. virescens, E. violaceus, and E. hyperarcticus (Poaceae: Triticeae) as determined by amplified fragment length polymorphism. AB - Morphological similarity among E. trachycaulus, E. virescens, E. violaceus, and E. hyperarcticus has often been noted. Taxonomists have tried to discriminate among these taxa using morphological characters and a number of different relationships among them have been suggested. However, the genetic relationships among these taxa are still unknown. AFLP analysis was used to characterize the molecular diversity of these taxa and to examine genetic relationships among them. A high degree of genetic identity was apparent among 7 accessions of E. virescens. The similarity values ranged from 0.90 to 0.99 with an average of 0.94. The mean similarity values among 3 E. hyperarcticus and among 5 E. violaceus accessions were 0.84 (0.81-0.87) and 0.77 (0.66-0.90), respectively. The similarity values among 17 E. trachycaulus accessions ranged from 0.49 to 0.92 with an average of 0.75. The 5 accessions of E. subsecundus displayed high variation, with similarity values between 0.52 and 0.68 and a mean value of 0.59. Both maximum-parsimony (MP) and neighbor-joining (NJ) analyses showed that all 7 accessions of E. virescens formed a clade, indicating a monophyletic origin. On the other hand, Elymus trachycaulus, E. subsecundus, and E. violaceus were each paraphyletic and separated into different genetically distinct groups. Among these 5 taxa, E. virescens was genetically similar to E. trachycaulus, and E. violaceus was genetically similar to E. hyperarcticus. PMID- 17110997 TI - Genetic relationships within and among Iberian fescues (Festuca L.) based on PCR amplified markers. AB - The genus Festuca comprises approximately 450 species and is widely distributed around the world. The Iberian Penninsula, with more than 100 taxa colonizing very diverse habitats, is one of its main centers of diversification. This study was conducted to assess molecular genetic variation and genetic relatedness among 91 populations of 31 taxa of Iberian fescues, based on several molecular markers (random amplified polymorphic DNA, amplified fragment length polymorphisms, and trnL sequences). The analyses showed the paraphyletic origin of the broad-leaved (subgenus Festuca, sections Scariosae and Subbulbosae, and subgenus Schedonorus) and the fine-leaved fescues (subgenus Festuca, sections Aulaxyper, Eskia, and Festuca). Schedonorus showed a weak relationship with Lolium rigidum and appeared to be the most recent of the broad-leaved clade. Section Eskia was the most ancient and Festuca the most recent of the fine-leaved clade. Festuca and Aulaxyper were the most related sections, in concordance with their taxonomic affinities. All taxa grouped into their sections, except F. ampla and F. capillifolia (section Festuca), which appeared to be more closely related to Aulaxyper and to a new independent section, respectively. Most populations clustered at the species level, but some subspecies and varieties mixed their populations. This study demonstrated the value in combining different molecular markers to uncover hidden genetic relationships between populations of Festuca. PMID- 17110998 TI - Novel events associated with phenotypic reversion of a P element mutant in Drosophila melanogaster. AB - Transposable P elements have been used extensively for Drosophila mutagenesis. While their mutagenic activity has long been recognized, the mechanisms by which P elements cause mutations are varied and not completely understood. We describe here an experiment to replace a P element at vestigial (vg) that caused a strong mutant phenotype (P[21-3]) with a P element (P[21]) known to produce a very weak phenotype when inserted at vg. In addition to testing the feasibility of P element replacements at vg, our investigation led to the production of 7 new vg alleles and 1 apparent second site suppressor. All the vg21-3 revertants that we recovered had a P element inserted into the first exon of vg at the same location and in the same orientation as the original element in vg21-3, providing a unique opportunity to study the mechanism of transposon mutagenesis. A majority of the revertants arose from a previously described event: internal deletion of P sequences, including the P promoter. In addition, 3 novel reversions of the vg21 3 wing phenotype were recovered. The wings of homozygous vg21r36 flies were normal. However, vg21r36 in combination with a deletion of the vg locus exhibited a strong mutant wing phenotype. This was surprising, because the P element insertion in vg21r36 was very similar to that found in the vg21 allele, which showed only slight nicking of the wings in combination with a deletion. In vg21r4, reversion was caused by a tandem insertion of P[21] and the original P[21 3] element present in vg21-3. Finally, the vg21r7 revertant had a P[21-3] insert at vg and 3 additional P elements elsewhere in the genome. We hypothesize that reversion in the 3 novel cases might be caused by P repressor produced by an element at vg or, in the case of vg21r7, elsewhere in the genome. This raises an interesting aspect of P element evolution. While P transposons produce mutations that might prove deleterious to their host, their success in invading the genome of D. melanogaster may be explained by their ability to silence those same mutations by a range of repressor-producing elements. PMID- 17110999 TI - SNP-based markers for discriminating olive (Olea europaea L.) cultivars. AB - A set of 11 polymorphic markers (1 cleaved amplified polymorphic sequence (CAPS), 2 sequence-characterized amplified regions (SCARs), and 8 single-nucleotide polymorphism (SNP)-derived markers) was obtained for olive cultivar identification by comparing DNA sequences from different accessions. Marker development was more efficient, using sequences from the database rather than cloning arbitrary DNA fragments. Analyses of the sequences of 3 genes from 11 diverse cultivars revealed an SNP frequency of 1 per 190 base pairs in exons and 1 per 149 base pairs in introns. Most mutations were silent or had little perceptible effect on the polypeptide encoded. The higher incidence of transversions (55%) suggests that methylation is not the major driving force for DNA base changes. Evidence of linkage disequilibrium in 2 pairs of markers has been detected. The set of predominantly SNP-based markers was used to genotype 65 olive samples obtained from Europe and Australia, and was able clearly to discriminate 77% of the cultivars. Samples, putatively of the same cultivar but derived from different sources, were revealed as identical, demonstrating the utility of these markers as tools for resolving nomenclature issues. Genotyping data were used for constructing a dendrogram by UPGMA cluster analysis using the simple matching similarity coefficient. Relationships between cultivars are discussed in relation to the route of olive's spread. PMID- 17111001 TI - Dietary intake, weight gain, and birth outcomes of physically active pregnant women: a pilot study. AB - Pregnant women have increased energy and nutrient needs. Exercise also increases energy and glucose demands. This pilot study examined whether women who were physically active during pregnancy met their dietary needs, gained weight adequately, avoided hypoglycemia, and had normal birth outcomes. Based on a convenience sample, 11 active women were studied from 18 weeks gestation to delivery. They exercised for 40 min at 70% VO2 peak, 3-4 times per week. Weekly measures included capillary blood glucose concentrations, body mass, and food records. Birth outcome measures included baby mass, length, and activity, pulse, grimace, appearance, and respiration (APGAR) scores. Our findings suggest that these subjects had lower average energy intake (8680 +/- 490 kJ/d) than recommended for the 2nd and 3rd trimesters (13,020 +/-185 kJ/d; 13,480 +/- 185 kJ/d, respectively, p < 0.0001). Protein requirements for pregnancy were exceeded. Average intakes of iron (19 +/- 2 mg/d) and folate (356 +/- 33 microg/d) were lower than Recommended Dietary Allowances (RDAs) (27 mg/d and 600 microg/d, respectively) and milk products consumed were less than the recommended daily servings. Four women gained less weight per week than recommended for pregnancy. No hypoglycemia occurred post-exercise. Birth outcome measures were within normal ranges. Comparative studies with a larger number of subjects are required to determine if active pregnant women have adequate nutrient intakes and weight gain. PMID- 17111002 TI - Creatine intake attenuates corticosteroid-induced impairment of voluntary running in hamsters. AB - Myopathy is a well-known side effect of corticosteroid therapy. Creatine monohydrate (Cr) supplementation has been shown to increase fat-free mass and muscular function. This study aimed to investigate if Cr administration could offset the deleterious functional effects of high doses of steroids. Fifty-six male Syrian golden hamsters were randomized among 4 groups: GI (n = 10), subcutaneous (s.c.) and intraperitoneal (i.p.) saline; GII (n = 10), s.c. saline and i.p. Cr (600 mg.kg(-1).d(-1)); GIII (n = 18), s.c. dexamethasone (7.5 mg.kg( 1).d(-1)) and i.p. saline; and GIV (n = 18), s.c. dexamethasone and i.p. Cr. Daily voluntary running was measured using activity wheels for 18 d. At the end of the study, statistically significant differences in running were observed between all groups, except for GI versus GII (GI, 8878 +/- 2737 m; GII, 9145 +/- 2000 m; GIII, 4289 +/- 2623 m; GIV, 6339 +/- 2345 m). Dexamethasone led to a significant decrease in cross-sectional area of type II fibers of the medial gastrocnemius. The cross-sectional area of type I fibers was significantly larger in GIV than in GIII. In conclusion, Cr administration attenuated the impairment of daily spontaneous running of hamsters receiving a high dose of corticosteroids. Additional research is needed to clarify the clinical implications of this finding. PMID- 17111003 TI - Increased muscle oxidative potential following resistance training induced fibre hypertrophy in young men. AB - Some evidence suggests that resistance training may lower relative muscle mitochondrial content via "dilution" of the organelle in a larger muscle fibre. Such an adaptation would reduce fatigue resistance, as well as compromise oxidative ATP synthesis and the capacity for fatty-acid oxidation. We investigated the effect of resistance training on mitochondrial enzymes of the citric acid cycle (citrate synthase; CS) and beta-oxidation (beta-hydroxyacyl CoA dehydrogenase; beta-HAD), as well as markers of the potential for glucose phosphorylation (hexokinase; HK) and glycolysis (phosphofructokinase; PFK). Twelve untrained men (21.9 +/- 0.5 y; 1.79 +/- 0.03 m; 83.2 +/- 3.2 kg) participated in a 12 week progressive resistance-training program. Muscle biopsies were taken from the vastus lateralis before (PRE) and after (POST) training. Training increased mean muscle fibre cross-sectional area (p < 0.05) and the activities of CS (PRE = 4.53 +/- 0.44 mol.kg protein(-1).h(-1); POST = 5.63 +/- 0.40 mol.kg protein(-1).h(-1); p < 0.001) and beta-HAD (PRE = 2.55 +/- 0.28 mol.kg protein(-1).h(-1); POST = 3.11 +/- 0.21 mol.kg protein(-1).h(-1); p < 0.05). The activity of HK increased 42% (p < 0.05), whereas the activity of PFK remained unchanged. We conclude that resistance training provides a stimulus for improving muscle oxidative potential, as reflected by the increased activities of CS and beta-HAD following resistance training induced hypertrophy. PMID- 17111004 TI - Acute effects of exercise timing and breakfast meal glycemic index on exercise induced fat oxidation. AB - Fat balance is an important determinant of energy balance. Exercise after an overnight fast can significantly increase fat oxidation; however, little information pertaining to the effects of exercise and meal glycemic index on fat oxidation under these conditions is available. The objective of this investigation was to study the acute effects of exercise timing and meal glycemic index (GI) on whole-body fat oxidation. Eight apparently healthy young men completed 4 randomly ordered trials during which measurements were made at rest, during exercise, and for 2 h post-exercise and (or) post-prandial. After an overnight fast, subjects were required to perform 400 kcal (1 kcal = 4.184 kJ) of treadmill exercise (at FATmax) either before consuming a 400 kcal low-GI (Ex-LG) or high-GI (Ex-HG) oatmeal breakfast, or after consuming the low-GI (LG-Ex) or high-GI (HG-Ex) meal. The amount of fat oxidized during exercise was significantly greater during Ex-LG and Ex-HG (17.2 +/- 4.0 and 17.5 +/- 4.7 g, respectively) than during LG-Ex and HG-Ex (10.9 +/- 3.7 and 11.7 +/- 3.5 g, respectively) (p < 0.001), as was the amount of fat oxidized during the entire trial (Ex-LG: 23.4 +/- 4.7 g; Ex-HG: 23.4 +/- 6.5 g; LG-Ex: 18.4 +/- 4.7 g; HG Ex: 19.6 +/- 4.9 g) (p < 0.05), even though energy expenditure was not different across experimental conditions. No significant effect of meal GI on the amount of fat oxidized was noted. Total fat oxidized during exercise, and for 2 h after exercise, was greatest when morning exercise was performed in the fasted state, independently of meal GI. PMID- 17111005 TI - Short-term lifestyle modification alters circulating biomarkers of endothelial health in sedentary, overweight adults. AB - Obesity and inactivity are associated with endothelial dysfunction that may contribute to the development of atherosclerosis. We examined the effects of a short-term lifestyle intervention on circulating biomarkers of endothelial health. Nineteen overweight or obese (mean body mass index (BMI): 28.9 +/- 0.7 kg/m2) men and women underwent 6 weeks of body mass reduction induced by moderate energy restriction (approximately 750 kcal/d; 1 kcal = 4.184 kJ) and aerobic training (approximately 400 kcal/d). Fasting serum samples were collected at baseline and after reduction in body mass (week 6) to assess concentrations of nitrotyrosine (NT), secretory phospholipase A2 (sPLA2), and soluble intracellular adhesion molecule-1 (sICAM-1). Body mass was significantly reduced from 81.3 +/- 2.8 to 77.3 +/- 2.6 kg (p < 0.05). Circulating concentrations of NT and sICAM-1 were significantly reduced with treatment (approximately 25% and approximately 10%, respectively), whereas sPLA2 levels were significantly elevated (approximately 45%). Elevations in sPLA2 were negatively correlated with changes in NT (r = -0.58, p = 0.047); reductions in NT did not correlate significantly with reductions in sICAM-1. It appears that circulating markers of endothelial health are susceptible to short-term exercise interventions with modest reduction in body mass, and such a lifestyle modification may improve endothelial health by reducing protein nitration products and cellular adhesion. PMID- 17111006 TI - Addition of glutamine to essential amino acids and carbohydrate does not enhance anabolism in young human males following exercise. AB - We examined the effect of a post-exercise oral carbohydrate (CHO, 1 g.kg(-1).h( 1)) and essential amino acid (EAA, 9.25 g) solution containing glutamine (0.3 g/kg BW; GLN trial) versus an isoenergetic CHO-EAA solution without glutamine (control, CON trial) on muscle glycogen resynthesis and whole-body protein turnover following 90 min of cycling at 65% VO2 peak. Over the course of 3 h of recovery, muscle biopsies were taken to measure glycogen resynthesis and mixed muscle protein synthesis (MPS), by incorporation of [ring-2H5] phenylalanine. Infusion of [1-13C] leucine was used to measure whole-body protein turnover. Exercise resulted in a significant decrease in muscle glycogen (p < 0.05) with similar declines in each trial. Glycogen resynthesis following 3 h of recovery indicated no difference in total accumulation or rate of repletion. Leucine oxidation increased 2.5 fold (p < 0.05) during exercise, returned to resting levels immediately post-exercise,and was again elevated at 3 h post-exercise (p < 0.05). Leucine flux, an index of whole-body protein breakdown rate, was reduced during exercise, but increased to resting levels immediately post-exercise, and was further increased at 3 h post-exercise (p < 0.05), but only during the CON trial. Exercise resulted in a marked suppression of whole-body protein synthesis (50% of rest; p < 0.05), which was restored post-exercise; however, the addition of glutamine did not affect whole-body protein synthesis post-exercise. The rate of MPS was not different between trials. The addition of glutamine to a CHO + EAA beverage had no effect on post-exercise muscle glycogen resynthesis or muscle protein synthesis, but may suppress a rise in whole-body proteolysis during the later stages of recovery. PMID- 17111007 TI - Physiological characteristics of the best Eritrean runners-exceptional running economy. AB - Despite their young age, limited training history, and lack of running tradition compared with other East African endurance athletes (e.g., Kenyans and Ethiopians), male endurance runners from Eritrea have recently attained important running successes. The purposes of our study were (i) to document the main physical and physiological characteristics of elite black Eritrean distance runners (n = 7; age: 22 +/- 3 years) and (ii) to compare them with those of their elite white Spanish counterparts. For this second purpose we selected a control group of elite Spanish runners (n = 9; 24 +/- 2 years), owing to the traditionally high success of Spanish athletes in long-distance running compared with other white runners, especially in cross-country competitions. The subjects' main anthropometric characteristics were determined, together with their maximum oxygen uptake (VO2 max) and VO2 (mL.kg(-1).min(-1)), blood lactate, and ammonia concentrations while running at 17, 19, or 21 km.h(-1). The body mass index (18.9 +/- 1.5 kg.m(-2)) and maximal calf circumference (30.9 +/- 1.5 cm) was lower in Eritreans than in Spaniards (20.5 +/- 1.7 kg.m(-2) and 33.9 +/- 2.0 cm, respectively) (p < 0.05 and p < 0.01, respectively) and their lower leg (shank) length was longer (44.1 +/- 3.0 cm vs. 40.6 +/- 2.7 cm, respectively) (p < 0.05). VO2 max did not differ significantly between Eritreans and Spaniards (73.8 +/- 5.6 mL.kg(-1).min(-1) vs. 77.8 +/- 5.7 mL.kg(-1).min(-1), respectively), whereas the VO2 cost of running was lower (p < 0.01) in the former (e.g., 65.9 +/- 6.8 mL.kg(-1).min(-1) vs. 74.8 +/- 5.0 mL.kg(-1).min(-1) when running at 21 km.h( 1)). Our data suggest that the excellent running economy of Eritreans is associated, at least partly, with anthropometric variables. Comparison of their submaximal running cost with other published data suggests that superior running economy, rather than enhanced aerobic capacity, may be the common denominator in the success of black endurance runners of East African origin. PMID- 17111008 TI - Verification phase as a useful tool in the determination of the maximal oxygen uptake of distance runners. AB - This study investigated the utility of a verification phase for increasing confidence that a "true" maximal oxygen uptake had been elicited in 16 male distance runners (mean age (+/-SD), 38.7 (+/- 7.5 y)) during an incremental treadmill running test continued to volitional exhaustion. After the incremental test subjects performed a 10 min recovery walk and a verification phase performed to volitional exhaustion at a running speed 0.5 km.h(-1) higher than that attained during the last completed stage of the incremental phase. Verification criteria were a verification phase peak oxygen uptake or= 3%) lower than the peak values attained in the incremental phase. Further research is required to improve the verification procedure before its utility can be confirmed. PMID- 17111009 TI - Maximum effort breath-hold times for males and females of similar pulmonary capacities during sudden face-only immersion at water temperatures from 0 to 33 degrees C. AB - For non breath-hold-trained males and females matched for pulmonary capacity and body size, the effects of sex, water temperature, and end-tidal gas tensions were studied for their potential influences on breath-holding ability. Maximum breath hold time (BHTmax) was measured a total of 546 times in 13 males and 13 females, each repeating 3 trials of sudden face immersion (i.e., no prior hyperventilation) in water at 0, 5, 10, 15, 20, and 33 degrees C and in an air control condition (AIR). End-tidal carbon dioxide (P(ET)CO2) and oxygen (P(ET)O2) gas tensions were measured before and after breath-holding in a subset of 11 males and 11 females. For BHTmax there was no main effect of sex (p = 0.20), but there was a main effect of immersion condition (p < 0.001). Relative to pre immersion rest values, end-tidal gas tensions were significantly higher in males than in females (p 120 mmHg), respectively. Endothelial dysfunction was assessed using isolated aortic ring, electron microscopy of thoracic aorta, and serum concentration of nitrite/nitrate. The expression of mRNA for p22phox and eNOS was assessed by using RT-PCR. Serum thiobarbituric acid reactive substances concentration and aortic superoxide anion concentration were estimated to assess oxidative stress. Fasudil (30 mg.kg-1, p.o.) and atorvastatin (30 mg.kg-1, p.o.) treatments markedly prevented hypercholesterolemia- and hypertension-evoked attenuation of acetylcholine-induced endothelium-dependent relaxation, impairment of vascular endothelial lining, decrease in expression of mRNA for eNOS and serum nitrite/nitrate concentration, and an increase in expression of mRNA for p22phox, superoxide anion, and serum thiobarbituric acid reactive substances. The ameliorative effect of fasudil was prevented by L-NAME. In conclusion, fasudil-induced inhibition of Rho-kinase may improve hypercholesterolemia- and hypertension-induced endothelial dysfunction. PMID- 17111029 TI - Selected plant species from the Cree pharmacopoeia of northern Quebec possess anti-diabetic potential. AB - Type II diabetes is a major health problem worldwide. Some populations, such as aboriginal peoples, are particularly at risk for this disease. In the Cree Nation of Quebec, Canada, prevalence in adults is approaching 20%, and the consequences are compounded by low compliance with modern medicine. In 2003, we conducted an ethnobotanical study of Cree medicinal plants used for the treatment of symptoms of diabetes. This served as the basis for a project designed to identify efficacious complementary treatment options more readily accepted by this population. The present study assesses the in vitro anti-diabetic potential of extracts from the 8 most promising plants to emerge from the ethnobotanical study. Cell-based bioassays were employed to screen for (i) potentiation of glucose uptake by skeletal muscle cells (C2C12) and adipocytes (3T3-L1); (ii) potentiation of glucose-stimulated insulin secretion (GSIS) and insulin production by pancreatic beta cells (INS 832/13); (iii) potentiation of triglyceride accumulation in differentiating 3T3-L1 cells; (iv) protection against glucose toxicity and glucose deprivation in pre-sympathetic neurons (PC12 AC). Additionally, anti-oxidant activity was measured biochemically by the diphenylpicrylhydrazyl (DPPH) reduction assay. All plant extracts potentiated basal or insulin-stimulated glucose uptake to some degree in muscle cells or adipocytes. Adipocyte differentiation was accelerated by 4 extracts. Five extracts conferred protection in PC12 cells. Three extracts displayed free radical scavenging activity similar to known anti-oxidants. None of the plant extracts enhanced GSIS or insulin content in INS 832/13 beta cells. It is concluded that the Cree pharmacopoeia contains several plants with significant anti-diabetic potential. PMID- 17111030 TI - Are there ventricle-specific postnatal maturational differences in myocardial beta-adrenoceptors? AB - Newborn hearts have restricted functional reserve and variable responsiveness to inotropes that could be partly due to differences in myocardial beta adrenoceptors (beta-AR). To clarify this issue, this study documented ventricle specific changes in myocardial beta-AR density and affinity during postnatal maturation. In vivo left and right ventricle (LV and RV, respectively) biopsies were obtained from newborn (3-day-old, n = 11), immature (14-day-old, n = 7), and adult (n = 6) pigs. Total beta-AR density (B(max), fmol/g) and dissociation constant (K(d), pmol/L) were determined by radioligand binding with I125 iodocyanopindolol. Overall, beta-AR B(max) in the LV significantly decreased with maturation. Interestingly, newborn animal hearts (LV and RV) subdivided into 2 groups: an adult-like low K(d) group with low B(max) and a fetal-like high K(d) group with high B(max), which were significantly different from one another. The high K(d) newborn group also had significantly higher K(d) and B(max) than both immature and adult hearts. Newborns had similar Bmax but higher Kd in the LV than the RV, whereas immature and adult hearts did not have ventricular differences. During maturation, beta-AR density decreased, whereas LV beta-AR binding affinity increased. Variable beta-AR maturity was also identified immediately post partum, which could potentially explain the newborn heart's variable responsiveness to inotropes. The subset of newborn hearts with lower binding affinity (reduced responsiveness) could also contribute to the newborn heart's overall reduction in functional reserve. PMID- 17111031 TI - Differential actin isoform reorganization in the contracting A7r5 cell. AB - In the present study, we investigated the reorganization of alpha- and beta-actin in the contracting A7r5 smooth muscle cell. The remodeling of these actin variants was markedly different in response to increasing concentrations of phorbol 12, 13-dibutyrate (PDBu). At the lowest concentrations (< or =10(-7) mol/L), cells showed an approximately 70% loss in alpha-actin stress fibers with robust transport of this isoform to podosomes. By comparison, beta-actin remained in stress fibers in cells stimulated at low concentrations (< or =10(-7) mol/L) of PDBu. However, at high concentrations (> or =10(-6)mol/L) approximately 50% of cells showed transport of beta-actin to podosomes. Consistent with these findings, staining with phalloidin indicated a significant decrease in the whole cell content of F-actin with PDBu treatment. However, staining with DNase I indicated no change in the cellular content of G-actin, suggesting reduced access of phalloidin to tightly packed actin in the podosome core. Inhibition of protein kinase C (staurosporine, bisindolymaleimide) blocked PDBu-induced (5 x 10(-8) mol/L) loss in alpha-actin stress fibers or reversed podosome formation with re establishment of alpha-actin stress fibers. By comparison, these inhibitors caused partial loss of beta-actin stress fibers. The results support our earlier conclusion of independent remodeling of alpha- and beta-actin cytoskeletal structure and suggest that the regulation of these structures is different. PMID- 17111032 TI - Increased severity of renal impairment in nephritic mice lacking the EP1 receptor. AB - In experimental glomerulonephritis, inhibition of renal prostaglandin (PG) synthesis by nonsteroidal-anti-inflammatory drugs (NSAIDs) moderates proteinuria, yet can induce harmful effects on renal blood flow and Na+ - K+ - water balance thereby implicating 1 or more prostanoid receptor subtypes. We investigated the role of the PGE2 EP1 receptor in nephritis since it is expressed in the glomerulus, collecting duct and vasculature in which its activity might contribute to adaptive or maladaptive responses. Accordingly, a mouse model of accelerated antiglomerular basement membrane (anti-GBM) nephrotoxic serum (NTS) nephritis was induced in mice with targeted-deletion of the EP1 receptor (EP1-/ ). Proteinuria was similar between wild-type (wt) and EP1-/- NTS groups, thus negating a role for this subtype in modulating the glomerular permeability barrier in this model of anti-GBM NTS. However, overall renal damage was more acute in NTS EP1-/- mice, as evidenced by the degree of glomerular mesangial matrix expansion and the frequency of tubular dilatations. These changes in renal pathology were accompanied by stronger impairment of renal function in NTS EP1-/- mice, such that levels of serum creatinine, urea, Na+, and K+ were each significantly higher than those observed in NTS wt mice. Lastly, compared with wt mice, induction of NTS more severely reduced urine osmolality and body mass in EP1-/- mice. Taken together, the increased renal impairment seen in NTS EP1-/- mice suggests that the EP1 subtype plays a compensatory role in the context of acute nephritis. PMID- 17111033 TI - Variations in parametrial white adipose tissue mass during the mouse estrous cycle: relationship with the expression of peroxisome proliferator-activated receptor-gamma and retinoic acid receptor-alpha. AB - Estrogen and progestin participate in the regulation of adipose tissue metabolism, and peroxisome proliferator-activated receptor-gamma (PPARgamma) and retinoic acid receptor-alpha (RXRalpha) are absolutely required for adipose tissue development. The present study is to investigate the changes in parametrial fat mass and expression of PPARgamma and RXRalpha during estrous cycle in mice. Parametrial white adipose tissues (WAT), inter-scapula brown adipose tissues, and uteri from female mice were weighed. Blood samples were collected for the measurement of 17 beta-estradiol and progesterone levels. An RNase protection assay and Western blot analysis were used to compare the expression of PPARgamma and RXRalpha in adipose tissue. The mass of parametrial WAT in diestrus was significantly higher compared with estrus. However, there is no significant difference on the mass of brown adipose tissues during estrous cycle. The expression of PPARgamma in WAT in diestrus was significantly higher than that in estrus. The expression of RXRalpha during estrous cycle was unchanged in both white and brown adipose tissues. In conclusion, the variation in parametrial WAT mass during the mouse estrous cycle correlates with changes in the expression of PPARgamma in WAT. PMID- 17111034 TI - Heme oxygenase activity in fetal and adult sheep is not altered by acclimatization to high altitude hypoxia. AB - Hypoxic stress has been reported to induce the expression of stress proteins such as heme oxygenase (HO), which catalyze the breakdown of heme to generate biliverdin, ferrous iron, and carbon monoxide. These degradation products play a role in the regulation of a variety of processes such as vascular tone, inflammation, and central nervous system function. In mammals, there are 2 catalytically functional HO isozymes, HO-1 (inducible) and HO-2 (constitutive). HO-1 expression is regulated by an array of nonphysiological and physiological stimuli including acute hypoxemia. As relatively little is known of the HO response to prolonged hypoxia in whole animals other than small laboratory rodents, the aim of this work was to examine the effect of long-term hypoxia on total HO activity in fetal and adult ovine tissue. Sheep were maintained at high altitude (3820 m), after which the following tissues were harvested from near term fetal and non-pregnant ewes for in vitro measurement of HO activity: left ventricle, renal papilla, lung apex, pulmonary artery, carotid artery, mesenteric artery, placental cotyledon, spleen, and brain frontal cortex. There were no significant differences between HO activities in tissues from hypoxic fetal and adult sheep compared with their normoxic controls. Fetal heart HO activities were higher than those of adult tissue (p < 0.05), whereas adult spleen HO activity was significantly higher than that of fetal tissue (p < 0.05). In conclusion, these data indicate that long-term exposure to high altitude hypoxia does not have a persistent effect on HO activity in ovine tissues. Also, except for the spleen where there is a high expression of HO-1 under normal conditions, tissue HO activity is correlated with the expression of HO-2, the constitutive isozyme. PMID- 17111035 TI - The effect of an adenosine and lidocaine intravenous infusion on myocardial high energy phosphates and pH during regional ischemia in the rat model in vivo. AB - We have previously shown that an intravenous infusion of adenosine and lidocaine (AL) solution protects against death and severe arrhythmias and reduces infarct size in the in vivo rat model of regional ischemia. The aim of this study was to examine the relative changes of myocardial high-energy phosphates (ATP and PCr) and pH in the left ventricle during ischemia-reperfusion using 31P NMR in AL treated rats (n = 7) and controls (n = 6). The AL solution (A: 305 microg.(kg body mass)-1.min-1; L: 608 microg.(kg body mass)-1.min-1) was administered intravenously 5 min before and during 30 min coronary artery ligation. Two controls died from ventricular fibrillation; no deaths were recorded in AL treated rats. In controls that survived, ATP fell to 73% +/- 29% of baseline by 30 min ischemia and decreased further to 68% +/- 28% during reperfusion followed by a sharp recovery at the end of the reperfusion period. AL-treated rats maintained relatively constant ATP throughout ischemia and reperfusion ranging from 95% +/- 6% to 121% +/- 10% of baseline. Owing to increased variability in controls, these results were not found to be significant. In contrast, control [PCr] was significantly reduced in controls compared with AL-treated rats during ischemia at 10 min (68% +/- 7% vs. 99% +/- 6%), at 15 min (68% +/- 10% vs. 93% +/ 2%), and at 20 min (67% +/- 15% vs. 103% +/- 5%) and during reperfusion at 10 min (56% +/- 22% vs. 99% +/- 7%), at 15 min (60% +/- 10% vs. 98% +/- 7%), and at 35 min (63% +/- 14% vs. 120% +/- 11%) (p < 0.05). Interestingly, changes in intramyocardial pH between each group were not significantly different during ischemia and fell by about 1 pH unit to 6.6. During reperfusion, pH in AL-treated rats recovered to baseline in 5 min but not in controls, which recovered to only around pH 7.1. There was no significant difference in the heart rate, mean arterial pressure, and rate-pressure product between the controls and AL treatment during ischemia and reperfusion. We conclude that AL cardioprotection appears to be associated with the preservation of myocardial high-energy phosphates, downregulation of the heart at the expense of a high acid-load during ischemia, and with a rapid recovery of myocardial pH during reperfusion. PMID- 17111036 TI - Amygdala adenosine A1 receptors have no anticonvulsant effect on piriform cortex kindled seizures in rat. AB - Adenosine is an endogenous anticonvulsant that exerts its effects through A1 receptors. As the piriform/amygdala is a critical circuit for limbic seizure propagation, in this study, the role of basolateral amygdala A1 receptors on piriform cortex (PC)-kindled seizures was investigated. Rats were kindled by daily electrical stimulation of PC. In fully kindled animals, bilateral intra amygdala N6-cyclohexyladenosine (CHA; 10-500 micromol/L, a selective A1 receptor agonist) had no effect on kindled-seizure parameters. However, bilateral intra amygdala 2% lidocaine (reversal neuronal inhibitor) reduced the kindled seizure severity. There was significant increase in stage 4 latency and decrease in stage 5 duration. Bilateral lesion of basolateral amygdala of kindled animals (by electrical DC current) reduced the kindled seizure severity more dramatically. Our results showed afterdischarge duration, stage 5 duration, and seizure duration were decreased and stage 4 latency increased significantly. In addition, daily intra-amygdala CHA had no significant effect on PC kindling acquisition. Therefore, it may be concluded that although the basolateral amygdala neuronal activity has a critical role in the propagation of epileptic seizures from PC, the amygdala A1 receptors have no role in this regard. On the other hand, amygdala A1 receptors have no anticonvulsant or antiepileptogenic effect on PC kindled seizures. PMID- 17111037 TI - Inhibition of polycystin-L channel by the Chinese herb Sparganum stoloniferum Buch.-Ham. AB - The Chinese herb Sparganum stoloniferum Buch.-Ham. (SBH) is frequently used to improve blood circulation and to rehabilitate vascular obstruction in traditional Chinese medicine. It was recently reported that SBH reduces the proliferation of renal epithelial cells stimulated by epidermal growth factor (EGF), and inhibits the phosphorylation of the EGF receptor. SBH has also been used as a trial drug to treat polycystic kidney disease (PKD) patients in China. The potential molecular actions of SBH on PKD remain unknown. Autosomal dominant PKD (ADPKD) is associated with mutations in polycystin-1 or polycystin-2 (PC2). PC2 and its homologue, polycystin-L (PCL), are nonselective cation channels permeable to potassium, sodium, and calcium. Here, we examine the effects of SBH on the human PCL channel expressed in Xenopus oocytes, using 2-microelectrode voltage-clamp electrophysiology and radiotracer uptake measurements. In PCL-expressing oocytes, with or without preincubation with SBH, the PCL channel was inhibited by SBH in a dose-dependent and reversible manner; a concentration of 2% SBH completely abolished the channel activation. The IC50 value for SBH was 0.48% +/- 0.03%, with a 10-min preincubation period. SBH was also found to inhibit the PCL mediated 45Ca tracer uptake in oocytes. Our study suggests that SBH contains 1 or more yet-to-be determined components that are inhibitors of PCL channel. The therapeutic potential of SBH for ADPKD and its chemical composition remain to be investigated. PMID- 17111038 TI - Whole-body heat shock protects the ischemic rat heart by stimulating mitochondria respiration. AB - Whole-body heat shock (HS) leads to an enhancement of postischemic mechanical function and an improvement in glucose use by the rat heart. Here, we examine the effect of HS on isolated mitochondrial metabolism during reperfusion in the working rat heart. Rats were anesthetized, and their body temperature was raised to 41-42 degrees C for 15 min. Control rats were treated the same way but were not exposed to hyperthermia. Twenty-four hours after HS or sham treatment, rats were reanesthetized and the hearts were removed for perfusion with Krebs Henseleit buffer, containing 11 mmol glucose/L and 1.2 mmol palmitate/L prebound to 3% albumin. Hearts were subjected to 25 min of global ischemia followed by 30 min of reperfusion. At the end of reperfusion, heart mitochondria were isolated using differential centrifugation and respiration measured in the presence of pyruvate, glutamate, or palmitoylcarnitine. Hearts subjected to HS showed an enhanced recovery of function, expressed as aortic flow, during the reperfusion period, compared with sham hearts. This improved functional status was associated with a significant increase in state 3 respiration in the presence of pyruvate, glutamate, or palmitoylcarnitine. These results show that HS offers protection against ischemic damage, and that a possible mechanism might be the enhanced myocardial metabolism of fuels. PMID- 17111039 TI - Effects of triiodo-thyronine on angiotensin-induced cardiomyocyte hypertrophy: reversal of increased beta-myosin heavy chain gene expression. AB - Thyroid hormone-induced cardiac hypertrophy is similar to that observed in physiological hypertrophy, which is associated with high cardiac contractility and increased alpha-myosin heavy chain (alpha-MHC, the high ATPase activity isoform) expression. In contrast, angiotensin II (Ang II) induces an increase in myocardial mass with a compromised contractility accompanied by a shift from alpha-MHC to the fetal isoform beta-MHC (the low ATPase activity isoform), which is considered as a pathological hypertrophy and inevitably leads to the development of heart failure. The present study is designed to assess the effect of thyroid hormone on angiotensin II-induced hypertrophic growth of cardiomyocytes in vitro. Cardiomyocytes were prepared from hearts of neonatal Wistar rats. The effects of Ang II and 3,3',5-triiodo-thyronine (T3) on incorporations of [3H]-thymine and [3H]-leucine, MHC isoform mRNA expression, PKC activity, and PKC isoform protein expression were studied. Ang II enhanced [3H] leucine incorporation, beta-MHC mRNA expression, PKC activity, and PKCepsilon expression and inhibited alpha-MHC mRNA expression in cardiomyocytes. T3 treatment prevented Ang II-induced increases in PKC activity, PKCepsilon, and beta-MHC mRNA overexpression and favored alpha-MHC mRNA expression. Thyroid hormone appears to be able to reprogram gene expression in Ang II-induced cardiac hypertrophy, and a PKC signal pathway may be involved in such remodeling process. PMID- 17111040 TI - Relationship between right ventricular wave speed and elastance in dogs. AB - Wave speed (c) must be known to separate forward- and backward-going waves during wave-intensity analysis, which measures the energy transported by the waves in the circulation. c is related to elastance; the present study was performed to measure right ventricular (RV) c during the cardiac cycle and to compare c with RV elastance. In 7 dogs, we measured right atrial, pulmonary arterial, pericardial and 2 RV pressures, and pulmonary arterial flow. A pulse generator was connected to the RV apex, and c was measured by determining the transit time between the 2 high-fidelity RV pressure transducers; the distance was measured roentgenographically. Eight sonomicrometry crystals were implanted in the RV endocardium to calculate RV volume and, thereby, elastance. RV c ranged from approximately 1 m/s during diastole to approximately 4 m/s during systole. Log log plots of c vs. elastance were linear. These slopes represent the power relationships between c and elastance and ranged from 0.30 to 0.56; for the combined data, it was 0.31. Given knowledge of c, forward- and backward-going waves can be identified and their energy quantitated. In the canine RV, c is approximately proportional to 1/3 the power of elastance: log c = 0.31.log E - 2.05. PMID- 17111041 TI - The effect of nicotine on in vitro placental perfusion pressure. AB - Cigarette smoking throughout pregnancy is associated with several negative outcomes, of which an increased incidence of intra-uterine growth restriction (IUGR) is most pronounced. Gestationally age-matched infants born to smoking mothers are, on average, 200 g lighter at birth, per pack smoked per day. The mechanisms and specific tobacco compounds responsible for the increased risk of IUGR among smokers have yet to be identified; however, it is widely accepted that smoking women have compromised placental perfusion throughout gestation due to the vasoconstricting effect of nicotine on uterine and placental blood vessels. Despite the universal acceptance of this theory, very little work has been completed to date examining the vasoactive properties of nicotine within the human placenta. The objective of this study was to determine the effect of nicotine on placental vascular function. Normal-term human placentae were obtained after elective cesarean sections. An in vitro placental perfusion system was used; increasing doses of nicotine (20-240 ng/mL) were added to either the maternal (n = 5) or fetal (n = 3) circulation. The basal feto-placental perfusion pressure was 39.87 +/- 4.3 mmHg and was not affected by nicotine. This finding supports the hypotheses that nicotine does not directly affect placental microvascular function and that any contribution to fetal growth restriction is likely at the level of placental function (i.e., amino acid transport) and (or) uterine vascular function. PMID- 17111042 TI - Improved wound healing of cutaneous sulfur mustard injuries in a weanling pig model. AB - OBJECTIVE: The objective was to examine the efficacy of several treatment regimens in improving wound healing of cutaneous sulfur mustard (HD) injuries. METHODS: Wound healing studies were conducted in weanling pigs. Superficial dermal HD injuries were debrided at 48 hours postexposure using an erbium-doped yttrium aluminum garnet (Er:YAG) laser, followed by application of a treatment adjunct. A variety of noninvasive bioengineering methods were conducted during the postsurgical observation period to examine the various cosmetic and functional aspects of the skin. Histopathology was performed at the end of each study (14 or 21 days postsurgery). RESULTS: As noted clinically, reepithelialization was nearly complete by 7 days postsurgery for many of the sites treated with petrolatum and scarlet red dressings. By 21 days, the skin elasticity of the petrolatum-dressed sites was not significantly different from that of sham-exposed skin. Upon dressing removal on postsurgery day 4, the neoepidermis of allograft- and thin film-dressed sites was partially removed, with resultant petechial hemorrhaging. Mean pathology scores for hydrocolloid dressed sites were significantly lower than those of untreated HD-exposed sites on postsurgery day 14. CONCLUSIONS: Care must be taken during bandage changes, and a nonadherent dressing that could be left in place for a longer period of time (eg, 7 days) would be beneficial. The use of cultured epithelial allograft material may have a potential role if grown on a completely nonadherent backing and left undisturbed for at least a week. Xeroform Petrolatum and Scarlet Red Ointment dressings are effective and inexpensive treatment adjuncts for HD injuries. PMID- 17111043 TI - Cardiovascular and cerebrovascular events in the randomized, controlled Alzheimer's Disease Anti-Inflammatory Prevention Trial (ADAPT). AB - OBJECTIVES: The Alzheimer's Disease Anti-inflammatory Prevention Trial (ADAPT) was designed to evaluate the conventional NSAID naproxen sodium and the selective COX-2 inhibitor celecoxib for primary prevention of Alzheimer's dementia (AD). On 17 December 2004, after the Adenoma Prevention with Celecoxib (APC) trial reported increased cardiovascular risks with celecoxib, the ADAPT Steering Committee suspended treatment and enrollment. This paper reports on cardiovascular and cerebrovascular events in ADAPT. DESIGN: ADAPT is a randomized, placebo-controlled, parallel chemoprevention trial with 1-46 mo of follow-up. SETTING: The trial was conducted at six field sites in the United States: Baltimore, Maryland; Boston, Massachusetts; Rochester, New York; Seattle, Washington; Sun City, Arizona; and Tampa, Florida. PARTICIPANTS: The 2,528 participants were aged 70 y and older with a family history of AD. INTERVENTIONS: Study treatments were celecoxib (200 mg b.i.d.), naproxen sodium (220 mg b.i.d.), and placebo. OUTCOME MEASURES: Outcome measures were deaths, along with nonfatal myocardial infarction (MI), stroke, congestive heart failure (CHF), transient ischemic attack (TIA), and antihypertensive treatment recorded from structured interviews at scheduled intervals. Cox proportional hazards regression was used to analyze these events individually and in several composites. RESULTS: Counts (with 3-y incidence) of participants who experienced cardiovascular or cerebrovascular death, MI, stroke, CHF, or TIA in the celecoxib-, naproxen-, and placebo-treated groups were 28/717 (5.54%), 40/713 (8.25%), and 37/1070 (5.68%), respectively. This yielded a hazard ratio (95% confidence interval [CI]) for celecoxib of 1.10 (0.67-1.79) and for naproxen of 1.63 (1.04-2.55). Antihypertensive treatment was initiated in 160/440 (47.43%), 147/427 (45.00%), and 164/644 (34.08%). This yielded hazard ratios (CIs) of 1.56 for celecoxib (1.26-1.94) and 1.40 for naproxen (1.12-1.75). CONCLUSIONS: For celecoxib, ADAPT data do not show the same level of risk as those of the APC trial. The data for naproxen, although not definitive, are suggestive of increased cardiovascular and cerebrovascular risk. PMID- 17111044 TI - Evolution and translation of research findings: from bench to where? AB - The credibility and replication of research findings evolve over time, as data accumulate. However, translation of postulated research promises to real-life biomedical applications is uncommon. In some fields of research, we may observe diminishing effects for the strength of research findings and rapid alternations of exaggerated claims and extreme contradictions--the "Proteus Phenomenon." While these phenomena are probably more prominent in the basic sciences, similar manifestations have been documented even in clinical trials and they may undermine the credibility of clinical research. Significance-chasing bias may be in part responsible, but the greatest threat may come from the poor relevance and scientific rationale and thus low pre-study odds of success of research efforts. Given that we currently have too many research findings, often with low credibility, replication and rigorous evaluation become as important as or even more important than discovery. Credibility, replication, and translation are all desirable properties of research findings, but are only modestly correlated. In this essay, I discuss some of the evidence (or lack thereof) for the process of evolution and translation of research findings, with emphasis on the biomedical sciences. PMID- 17111045 TI - ADAPT: the wrong way to stop a clinical trial. PMID- 17111046 TI - Indoleamine 2,3-dioxygenase-expressing dendritic cells form suppurative granulomas following Listeria monocytogenes infection. AB - Control of pathogens by formation of abscesses and granulomas is a major strategy of the innate immune system, especially when effector mechanisms of adaptive immunity are insufficient. We show in human listeriosis that DCs expressing indoleamine 2,3-dioxygenase (IDO), together with macrophages, are major cellular components of suppurative granulomas in vivo. Induction of IDO by DCs is a cell autonomous response to Listeria monocytogenes infection and was also observed in other granulomatous infections with intracellular bacteria, such as Bartonella henselae. Reporting on our use of the clinically applied anti-TNF-alpha antibody infliximab, we further demonstrate in vitro that IDO induction is TNF-alpha dependent. Repression of IDO therefore might result in exacerbation of granulomatous diseases observed during anti-TNF-alpha therapy. These findings place IDO(+) DCs not only at the intersection of innate and adaptive immunity but also at the forefront of bacterial containment in granulomatous infections. PMID- 17111047 TI - Functional validation of the anaplastic lymphoma kinase signature identifies CEBPB and BCL2A1 as critical target genes. AB - Anaplastic large cell lymphomas (ALCLs) represent a subset of lymphomas in which the anaplastic lymphoma kinase (ALK) gene is frequently fused to the nucleophosmin (NPM) gene. We previously demonstrated that the constitutive phosphorylation of ALK chimeric proteins is sufficient to induce cellular transformation in vitro and in vivo and that ALK activity is strictly required for the survival of ALK-positive ALCL cells. To elucidate the signaling pathways required for ALK-mediated transformation and tumor maintenance, we analyzed the transcriptomes of multiple ALK-positive ALCL cell lines, abrogating their ALK mediated signaling by inducible ALK RNA interference (RNAi) or with potent and cell-permeable ALK inhibitors. Transcripts derived from the gene expression profiling (GEP) analysis uncovered a reproducible signature, which included a novel group of ALK-regulated genes. Functional RNAi screening on a set of these ALK transcriptional targets revealed that the transcription factor C/EBPbeta and the antiapoptotic protein BCL2A1 are absolutely necessary to induce cell transformation and/or to sustain the growth and survival of ALK-positive ALCL cells. Thus, we proved that an experimentally controlled and functionally validated GEP analysis represents a powerful tool to identify novel pathogenetic networks and validate biologically suitable target genes for therapeutic interventions. PMID- 17111048 TI - Myeloid progenitors differentiate into microglia and promote vascular repair in a model of ischemic retinopathy. AB - Vision loss associated with ischemic diseases such as retinopathy of prematurity and diabetic retinopathy are often due to retinal neovascularization. While significant progress has been made in the development of compounds useful for the treatment of abnormal vascular permeability and proliferation, such therapies do not address the underlying hypoxia that stimulates the observed vascular growth. Using a model of oxygen-induced retinopathy, we demonstrate that a population of adult BM-derived myeloid progenitor cells migrated to avascular regions of the retina, differentiated into microglia, and facilitated normalization of the vasculature. Myeloid-specific hypoxia-inducible factor 1alpha (HIF-1alpha) expression was required for this function, and we also demonstrate that endogenous microglia participated in retinal vascularization. These findings suggest what we believe to be a novel therapeutic approach for the treatment of ischemic retinopathies that promotes vascular repair rather than destruction. PMID- 17111049 TI - Glueing of fundal varices. PMID- 17111050 TI - Why adefovir is not yet available in Canada. PMID- 17111051 TI - Out in left field? A speculative viewpoint on the future. PMID- 17111052 TI - Commonly used preparations for colonoscopy: efficacy, tolerability, and safety--a Canadian Association of Gastroenterology position paper. AB - INTRODUCTION: The increased demand for colonoscopy, coupled with the introduction of new bowel cleansing preparations and recent caution advisories in Canada, has prompted a review of bowel preparations by the Canadian Association of Gastroenterology. METHODS: The present review was conducted by the Clinical Affairs group of committees including the endoscopy, hepatobiliary/transplant, liaison, pediatrics, practice affairs and regional representation committees, along with the assistance of Canadian experts in the field. An effort was made to systematically assess randomized prospective trials evaluating commonly used bowel cleansing preparations in Canada. RESULTS: Polyethylene glycol (PEG)-; sodium phosphate (NaP)-; magnesium citrate (Mg-citrate)-; and sodium picosulphate, citric acid and magnesium oxide (PSMC)-containing preparations were reviewed. Regimens of PEG 2 L with bisacodyl (10 mg to 20 mg) or Mg-citrate (296 mL) are as effective as standard PEG 4 L regimens, but are better tolerated. NaP preparations appear more effective and better tolerated than standard PEG solutions. PSMC has good efficacy and tolerability but head-to-head trials with NaP solutions remain few, and conclusions equivocal. Adequate hydration during preparation and up to the time of colonoscopy is critical in minimizing side effects and improving bowel cleansing in patients receiving NaP and PSMC preparations. All preparations may cause adverse events, including rare, serious outcomes. NaP should not be used in patients with cardiac or renal dysfunction (PEG solution is preferable in these patients), bowel obstruction or ascites, and caution should be exercised when used in patients with pre-existing electrolyte disturbances, those taking medications that may affect electrolyte levels and elderly or debilitated patients. Health Canada's recommended NaP dosing for most patients is two 45 mL doses 24 h apart. However, both safety and efficacy data on this dosing schedule are lacking. Many members of the Canadian Association of Gastroenterology expert panel administer both doses within 24 h, as studied in clinical trials, after careful one-on-one discussion of risks and benefits in carefully selected patients. Safety data on PSMC and combination preparations in North America are limited and clinicians are encouraged to keep abreast of developments in this area. CONCLUSIONS: All four preparations reviewed provided effective bowel cleansing for colonoscopy in the majority of patients, with varying tolerability. Adequate hydration is essential in patients receiving the preparations. PMID- 17111053 TI - Colorectal cancer screening: physicians' knowledge of risk assessment and guidelines, practice, and description of barriers and facilitators. AB - BACKGROUND: Physician nonadherence to colorectal cancer (CRC) screening recommendations contributes to underuse of screening. OBJECTIVE: To assess physicians' knowledge of CRC screening guidelines for average-risk individuals, perceived barriers to screening and practice behaviours. METHODS: Between October 2004 and March 2005, staff physicians working in three university-affiliated hospitals in Montreal, Quebec, were surveyed. Self-administered questionnaires assessed knowledge of risk classification and current guidelines for average-risk individuals, as well as perceptions of barriers to screening and practice behaviours. RESULTS: All 65 invited physicians participated in the survey, including 46 (70.8%) family medicine physicians and 19 (29.2%) general internists. Most physicians knew that screening should begin at 50 years of age, all knew to screen men and women and 92% said they screened average-risk patients. Fifty-seven (87.7%) physicians correctly identified three common characteristics associated with high risk for developing CRC. Physicians who screened average-risk patients preferred fecal occult blood testing (88.3%) and colonoscopy (88.3%) to flexible sigmoidoscopy (10.0%) and double-contrast barium enema (30.0%). Most physicians knew the correct screening periodicity for fecal occult blood testing (87.6%), but only 40% or fewer could identify correct screening periodicities for the other modalities. Barriers and facilitators focused on health care delivery system improvements, better evidence on which to base recommendations and development of practical screening modalities. CONCLUSIONS: Physicians lacked knowledge of the recommended screening modalities and periodicities to appropriately screen average-risk individuals. Because CRC screening can reduce mortality, efforts to improve physician delivery should focus on physician knowledge and changes to the health care delivery system. PMID- 17111054 TI - Gluten intolerance: sex and age-related features. AB - OBJECTIVE: Gluten intolerance is an immune-mediated enteropathy associated with gluten-containing foods in genetically susceptible patients. The typical form mainly affecting children shows failure to thrive and/or gastrointestinal symptoms. The adult form is less typical, presenting vague gastrointestinal symptoms, iron deficiency (with or without anemia) or nonspecific serum chemistry abnormalities. The present study aims to analyze clinical and biochemical differences of celiac disease (CD) according to sex and age. PATIENTS AND METHODS: The present study reviewed clinical and biochemical features of patients with suspected CD admitted to the Hospital General of Mostoles (Madrid, Spain) between July 2001 and June 2005. Two hundred fifty-two patients were analyzed, in whom intestinal biopsy was performed due to clinical and/or biochemical abnormalities suggestive of CD. One hundred seventy-eight asymptomatic relatives of the affected patients were also included. Overall, 125 patients showed diagnostic features of CD in the intestinal biopsy. RESULTS: The results confirmed higher prevalence of typical forms of CD in children (67% in children compared with only 14.3% in adults). CD seemed to be more frequent in adult women than in men (ratio of women to men 4:1), but it is worth noting that men diagnosed were most often referred with a typical clinical picture, so atypical forms of the disease in men may have been underdiagnosed. CONCLUSIONS: CD shows atypical features in adults, and physicians must include this disorder in the differential diagnosis of adults with iron deficiency or slight hypertransaminasemia. Increased awareness of the disease and extensive availability of accurate sero-logical tests will lead to improved diagnosis of this disorder, both in children and adults. PMID- 17111055 TI - Recurrent hepatitis C post-transplantation: where are we now and where do we go from here? A report from the Canadian transplant hepatology workshop. AB - Approximately 400 liver transplants are performed in Canada every year and close to 6000 per year in the United States. Forty per cent to 45% of all liver transplants are performed for patients with underlying hepatitis C virus (HCV) related liver disease. These patients have a different natural history, new complication risks and different treatment efficacy than nontransplant HCV patients. Every effort must be made to identify those patients at highest risk for progressive liver disease post-transplant. Recurrent HCV is an Achilles' heel to transplant hepatology. The true natural history of this disease is only starting to unravel and many questions remain unanswered on the optimal management of these patients after liver transplantation. The present report summarizes the literature and ongoing research needs that are specific to HCV related liver transplantation. PMID- 17111056 TI - New treatment strategies against hepatitis C viral infection. AB - Treatment of hepatitis C virus infection is currently based on a combination of pegylated interferon and ribavirin. Because efficacy of this therapy remains suboptimal and side effects sometimes problematic, major efforts have been put forward by scientists and the pharmaceutical industry to develop alternative treatments for this chronic infection. Over the past few years, clinical studies performed with some of these new agents have been presented at major international meetings. The present paper aims to review the rationale underlying the development of these new forms of treatment as well as the current available data concerning their clinical efficacy. PMID- 17111057 TI - [Canadian mental health rights in an international perspective]. AB - This article surveys the status of people with mental disorders in the light of international human rights law and assesses if their rights are respected in the Canadian context. The authors recognize that although the national systems of countries such as Canada provide significant civil and constitutional protections on the positive rights of its citizens, including those who suffer from intellectual disability, the same cannot be said with respect to entitlements to the provision of social services. The authors argue that this shortcoming must be remedied. Finally, the authors conclude that it is paramount to closely monitor the apparent dissonance between internationally recognized rights to adequate healthcare and freedom from discrimination and their strict application in the Canadian context. PMID- 17111058 TI - [Ethical deliberation: from case study to responsible citizenship]. AB - This article wants to be a contribution reflecting on the theme of applied ethics aiming at nourishing professional and interdisciplinary practices. The authors examine the process of ethical deliberation which has four major ends : the development of reflective and ethical competences of professionals, the elaboration by professionals, of an ethical reflective structure facilitating professional praxis, orientation and quick coordination of action ; implementation of a structure of collective deliberation useful in the orientation of difficult decisions and future actions. Finally, this process also has an objective of evaluation and appears relevant in the apprenticeship drawn from experiences of each participant. PMID- 17111059 TI - [Paths of subjects in the analytical course]. AB - During the analytical course, not necessarily at the end of the course of treatment, but as a resort to analysis, at the least as a first step, in a moment or another during the analytical process, the subject is called to take an ethical position. The stakes are obviously not the same in the beginning, at a decisive turning point or again at the end of treatment, meaning its conclusion. However, the subject cannot avoid this confrontation where self and self alone, decide to engage or not in the discovery of his unconscious. Clinical cases illustrate these turning points. As for the analyst, not giving up on his desire to know, he maintains the ethics of the psychoanalysis by remaining on the side of sharpness of truth and he guides his action in order for the subject to have access to its cause. PMID- 17111060 TI - [Taking the responsibility of a 'knowledge' one only has access during mental health consultations: stakes of an ethical issue]. AB - Everyone has a certain knowledge of what psychologically constitutes him/her. Part of the therapist's ethical approach is to help this knowledge come to conscience. Apart from the analytical process, within one or a few therapeutic sessions, is the person consulting able to have access to a minimal knowledge about him-herself? Using this knowledge to guide certain aspects of his-her life, calls upon an ethics of responsibility on the part of the person consulting. To have this knowledge emerge demands on the therapists' part, a concept of the human being that takes into account subjective dimensions of the person. PMID- 17111061 TI - [Clinical ethics in psychiatry: the experience at Douglas Hospital]. AB - The authors present a brief overview of the clinical ethics committee within their mental health university institute as well as its evolving mandates over the past few decades. The main functions include case consultation, input on elaboration of institutional policy, and staff education as well as public information on issues of ethics and mental illness. With examples and questions brought to the committee's attention, the authors demonstrate how these functions are intertwined. The authors conclude that ethical questioning helps examine clinical practices and serves ultimately in guiding towards best practices in mental health. PMID- 17111062 TI - [Clinical and ethical stakes of external supervision of mental health teams]. AB - External supervision of teams is increasingly part of mechanisms put in place both in public networks and community organizations to contribute in supporting professional mental health workers in developing their abilities and improving their clinical practices. It also plays a role in the analysis of ethical questions and in the prevention of deontological errors. The supervision of teams also includes its share of challenges, notably with regards to group dynamics and mechanisms to favour in order for supervision to rightly adjust to the needs and levels of participating professionals. Finally, the supervisor must also consider his own ethical responsibilities towards those supervised and towards their clients. PMID- 17111063 TI - [Psychoeducational and cognitive behavioral treatments of bipolar disorder]. AB - Bipolar disorder is a severe mood disorder characterized by recurrence of mania and depression. Despite the use of mood stabilizers, a significant proportion of bipolar patients experience relapse, psychosocial impairment and persistent symptoms. A significant part of patients show poor adhesion to the pharmacological treatment. This article aims to provide an overview of research focusing on psychoeducational and cognitive-behavioral treatment (CBT) of bipolar patients. METHOD: Studies were identified through Medline searches between 1971 and 2005. RESULTS: Studies on bipolar patients suggest that psychoeducational interventions may improve treatment adherence, illness knowledge, ability to cope with early manic symptoms and tend to reduce the risk of manic relapses. CBT tends to diminish depressive symptoms, improve treatment adherence and reduce the risk of depressive and manic relapses. Most psychoeducational and CBT studies share a common medical model of the illness, thereby making clear distinctions of impact of each intervention difficult. Few studies focused on patients with problems with mood stabilizers adherence. It is now important to develop specific interventions for those patients. CONCLUSION: According to these studies, bipolar patients are likely to benefit from psychoeducational or CBT interventions added to usual pharmacotherapy. In order to overcome limitations of existing research, future studies should adjust for the effect of pharmacological treatment, the type and severity of psychopathology at baseline, the acceptance of and the adaptability to the illness and it's awareness. PMID- 17111064 TI - [The process followed by helping peers during their experience with suicidal teenagers: an exploratory study]. AB - The purpose of this qualitative, exploratory and retrospective study is to understand the process followed by helping peers during their experience with a suicidal teenager. Interviews allowed to collect data with young people who had played this role. An analysis using grounded theory gave the following proposal: for youths, their experience was perceived as a heroic mission, first stimulating, then confronting and, finally, enriching. The experience goes through three stages. The first begins with the access to the status of helping peer and is pervaded with the wildly enthusiastic attitude of youth who give themselves the mission to save a suicidal mate. The second is marked by a certain disappointment having considered the facts surrounding the mission and undertaken a fight to save the suicidal mate. At the last stage, after the mission is completed, an attitude of wisdom stands out when the helping peer makes the synthesis of his victories and his defeats. The scarcity of studies on such a controversial subject incites to recommend further research. PMID- 17111065 TI - [Group therapy for men who are violent towards their spouse: dropout from treatment]. AB - It is now known that group treatment dropout rate for men who are violent towards their spouse constitutes a problem. The goal of this study is to verify if a link exists between attrition in a treatment for violent men and both partners's personal (age, revenue, substance abuse, having been subjected to violence as children) and marital variables (dyadic adjustment, anger, attribution, attachment style and violent behavior). Eighty men enrolled in a group treatment for marital violence have been recruited. Correlational analysis showed us that age is the only variable correlated with treatment dropout. The younger the participants, the most likely they were to drop out of the group therapy. These results will be further discussed. PMID- 17111066 TI - [Differences and similarities in perception of schizophrenia between physicians and the general population in Quebec]. AB - This paper presents results concerning the perceptions and attitudes of Quebec physicians towards patients with schizophrenia and compares data obtained from a previous poll to data drawn from answers of five common questions asked to the general population. A short questionnaire with 5 items selected earlier from a broader questionnaire submitted to the general population, has been distributed to Quebec physicians. These items inquired about the perceptions and attitudes of physicians towards schizophrenia. A randomized sample of physicians was performed. Three thousand and five hundred (3 500) physicians were selected and distributed questionnaires. A response rate of 29 %, a little more than one thousand (1003 responses) was observed, 46 % women and 54 % men. The authors have found significant differences between physicians and the general population in the tendency of wanting to offer help to those suffering from schizophrenia (physicians = 58 % versus general population : 45 %). Also, a higher percentage of physicians (72 %) have expressed feelings of compassion towards patients with schizophrenia versus 27 % in the general population. Results indicate that physicians, with a family member suffering from schizophrenia, are less comfortable discussing openly about the family member's illness (26 % versus 48 %). With regards to preconception of the severity of schizophrenia, in the field of health, and more specifically mental health, there are no differences observed amongst the physicians and the general population. PMID- 17111067 TI - Genetic and epigenetic risks of intracytoplasmic sperm injection method. AB - Pregnancies achieved by assisted reproduction technologies, particularly by intracytoplasmic sperm injection (ICSI) procedures, are susceptible to genetic risks inherent to the male population treated with ICSI and additional risks inherent to this innovative procedure. The documented, as well as the theoretical, risks are discussed in the present review study. These risks mainly represent that consequences of the genetic abnormalities underlying male subfertility (or infertility) and might become stimulators for the development of novel approaches and applications in the treatment of infertility. In addition, risks with a polygenic background appearing at birth as congenital anomalies and other theoretical or stochastic risks are discussed. Recent data suggest that assisted reproductive technology might also affect epigenetic characteristics of the male gamete, the female gamete, or might have an impact on early embryogenesis. It might be also associated with an increased risk for genomic imprinting abnormalities. PMID- 17111068 TI - Repair of necrosis and defects of penile skin with autologous free skin flap. AB - We described a 27-year-old case of avulsion and traumatic degloving of penile with extensive penis skin necrosis. Under general anesthesia, donor skin was partially resected from lower limbs according to defect area of penile skin. Then shear the shape of graft was sheared, sutured to hostage skin defect and enswathed with tension. The posto-perative appearance and function of the penis were satisfactory. It is suggest the homologous free skin flap from lower limbs is suitable for penile skin repair and beneficial to patient resulting in satisfactory erection and shape. PMID- 17111069 TI - Use of image analysis techniques for objective quantification of the efficacy of different hair removal methods. AB - In the field of consumer-used cosmetics for hair removal and hair growth reduction, there is a need for improved quantitative methods to enable the evaluation of efficacy and claim support. Optimized study designs and investigated endpoints are lacking to compare the efficacy of standard methods, like shaving or plucking, with new methods and products, such as depilating instruments or hair-growth-reducing cosmetics. Non-invasive image analysis, using a high-performance microscope combined with an optimized image analysis tool, was investigated to assess hair growth. In one step, high-resolution macrophotographs of the legs of female volunteers after shaving and plucking with cold wax were compared to observe short-term hair regrowth. In a second step, images obtained after plucking with cold wax were taken over a long-term period to assess the time, after which depilated hairs reappeared on the skin surface. Using image analysis, parameters like hair length, hair width, and hair projection area were investigated. The projection area was found to be the parameter most independent of possible image artifacts such as irregularities in skin or low contrast due to hair color. Therefore, the hair projection area was the most appropriate parameter to determine the time of hair regrowth. This point of time is suitable to assess the efficacy of different hair removal methods or hair growth reduction treatments by comparing the endpoint after use of the hair removal method to be investigated to the endpoint after simple shaving. The closeness of hair removal and visible signs of skin irritation can be assessed as additional quantitative parameters from the same images. Discomfort and pain rating by the volunteers complete the set of parameters, which are required to benchmark a new hair removal method or hair-growth-reduction treatment. Image analysis combined with high-resolution imaging techniques is a powerful tool to objectively assess parameters like hair length, hair width, and projection area. To achieve reliable data and to reduce well known image-analysis artifacts, it was important to optimize the technical equipment for use on human skin and to improve image analysis by adaptation of the image-processing procedure to the different skin characteristics of individuals, like skin color, hair color, and skin structure. PMID- 17111070 TI - Cosmeceutical properties of polysaccharides from the root bark of Ulmus davidiana var. japonica. AB - In Korea and China, Ulmus davidiana var. japonica has been used as a traditional oriental medicine for the treatment of difficulty in urination, skin inflammation, etc. In order to investigate the potential of a polysaccharide extract from Ulmus davidiana var. japonica as a cosmetic ingredient, we measured its moisturizing effect, photo-induced cytotoxicity, and anti-inflammatory effect. After hydrolysis, HPLC experiments showed that the composition of the polysaccharide extract was mainly rhamnose, galactose, and glucose. The molecular weight of the obtained Ulmus davidiana root extract was 20,000. The intrinsic viscosity was 90 dl/g. In a moisturizing test conducted through the measurement of water loss in a desiccator and of moisture content with a Corneometer CM820, Ulmus davidiana root extract showed almost the same moisturizing effect as hyaluronic acid. In an assay for inhibition of the H(2)O(2)-activated release of PGE2, IL-6, and IL-8 in normal human fibroblast cell lines, Ulmus davidiana root extract showed an inhibitory activity of PGE2 release in a dose-dependent manner (up to 85.9% at a concentration of 0.1%). The percent inhibition of the release of IL-6 was in the range of 45.6% to 64.5% (H(2)O(2) was used as the positive control). Moreover, the release of IL-8 was completely inhibited in the entire concentration range (>0.0025%). In a test of recovery from photo-induced damage after UVA irradiation (3 J/cm(2)), the cell recovery of human fibroblasts increased to levels two times higher than that of the positive control, which was UVA-damaged cells in the absence of Ulmus davidiana root extract (up to 60.2% at 3.0% of Ulmus davidiana root extract). In a photo-induced cytotoxicity assay in the presence of promethazine as a photosensitizer, Ulmus davidiana root extract showed approximately 48% of the increased cell viability of the control. Therefore, Ulmus davidiana root extract may be useful for the development of a cosmetic ingredient. PMID- 17111071 TI - A horse chestnut extract, which induces contraction forces in fibroblasts, is a potent anti-aging ingredient. AB - Contraction forces generated by non-muscle cells, such as fibroblasts, play important roles in determining cell morphology, vasoconstriction, and/or wound healing. We have searched among various plant extracts for ingredients that generate cell contraction forces using fibroblast-populated collagen gels. Using that model, we found that an extract of horse chestnuts (Aesculus hippocastanum) is able to generate such contraction forces in fibroblasts. The involvement of stress fiber formation in that response is suggested by the inhibition of such force generation by cytochalasin D and rhodamine phalloidin stain. Clinical testing of the extract was carried out using 40 healthy female volunteers. A gel formulation that included 3% of the extract was applied topically to the skin around the eye three times daily for nine weeks. The efficacy of the extract to diminish wrinkles was evaluated by visual scoring based on photo scales. After six weeks, significant decreases in the wrinkle scores at the corners of the eye or in the lower eyelid skin were observed compared with controls. After nine weeks, similar results were obtained. Taken together, our results suggest that an extract of horse chestnuts can generate contraction forces in fibroblasts and is a potent anti-aging ingredient. PMID- 17111072 TI - A novel volumetric method for quantitation of titanium dioxide in cosmetics. AB - Nowadays there are many sun-protection cosmetics incorporating organic or inorganic UV filters as active ingredients. Chemically stable inorganic sunscreen agents, usually metal oxides, are widely employed in high-SPF (sun protection factor) products. Titanium dioxide is one of the most frequently used inorganic UV filters. It has been used as a pigment for a long period of cosmetic history. With the development of micronization techniques, it has become possible to incorporate titanium dioxide in sunscreen formulations without the previous whitening effect, and hence its use in cosmetics has become an important research topic. However, there are very few works related to quantitation of titanium dioxide in sunscreen products. In this research, we analyzed the amounts of titanium dioxide in sunscreen cosmetics by adapting redox titration, reduction of Ti(IV) to Ti(III), and reoxidation to Ti(IV). After calcification of other organic ingredients of cosmetics, titanium dioxide is dissolved by hot sulfuric acid. The dissolved Ti(IV) is reduced to Ti(III) by adding metallic aluminum. The reduced Ti(III) is titrated against a standard oxidizing agent, Fe(III) (ammonium iron(III) sulfate), with potassium thiocyanate as an indicator. In order to test the accuracy and applicability of the proposed method, we analyzed the amounts of titanium dioxide in four types of sunscreen cosmetics, namely cream, make-up base, foundation, and powder, after adding known amounts of titanium dioxide (1 approximately 25 w/w%). The percentages of titanium dioxide recovered in the four types of formulations were in the range between 96% and 105%. We also analyzed seven commercial cosmetic products labeled with titanium dioxide as an ingredient and compared the results with those obtained from ICP-AES (inductively coupled plasma-atomic emission spectrometry), one of the most powerful atomic analysis techniques. The results showed that the titrated amounts were well in accord with the analyzed amounts of titanium dioxide by ICP-AES. Although instrument-based analytical methods, namely ICP-MS (inductively coupled plasma-mass spectrometry) and ICP-AES, are best for the analysis of titanium, it is difficult for small cosmetic companies to install such instruments because of their high cost. It was found that the volumetric method presented here gives quantitatively accurate and reliable results with routine lab-ware and chemicals. PMID- 17111073 TI - Photostabilization of organic UV-absorbing and anti-oxidant cosmetic components in formulations containing micronized manganese-doped titanium oxide. AB - Micronized titanium oxide (TiO(2)) and manganese-doped titanium oxide (TiO(2):Mn) particles have been incorporated into a variety of oil-in-water (O/W) and water in-oil-in-water (W/O/W) emulsions in conjunction with the UV-absorbing organic compounds butyl methoxydibenzoylmethane (BMDM) and octyl methoxycinnamate (OMC) and with the anti-oxidants vitamin E and vitamin C. The retention of the organics under solar exposure has been shown to be significantly enhanced by the addition of TiO(2):Mn to the formulation. In the case of BMDM and OMC, the retention is increased from 20% and 24% to 63% and 83%, respectively, after two hours of solar exposure. In this system, TiO(2) particles are shown to provide only limited protection relative to BMDM and OMC. Vitamin E and vitamin C are actively degraded by the presence of TiO(2) in the emulsion during solar exposure. This effect is reversed with TiO(2):Mn, the use of which can protect >90% of anti oxidants in both the oil and water phases of the formulation. The absence of reactive oxygen species (ROS) generation and surface scavenging of ROS by TiO(2):Mn is responsible for a significantly reduced ROS load on the organic components and consequent photostabilization of the emulsion. PMID- 17111077 TI - The correlation between serum prostate specific antigen levels and asymptomatic inflammatory prostatitis. AB - INTRODUCTION: Asymptomatic inflammatory prostatitis is a common pathological finding in patients with benign prostatic hyperplasia (BPH). In the present study, we evaluated the correlation between serum prostate specific antigen (PSA) level and extent and aggressiveness of inflammation in surgical specimens of patients who underwent to surgery for BPH without any evidence of clinical prostatitis. METHODS: Histological sections of the prostatic tissues of 52 patients were scored for the extent of inflammation and aggressiveness of inflammation, using the four point scale designed by Irani et al. Extent of inflammation is graded from 0 to 3 according to the degree of invasion of inflammatory cells in prostate tissue. Aggressiveness of inflammation is graded from 0 to 3 according to the degree of contact or disruption of prostatic glandular epithelium by inflammatory cells. The serum PSA levels in different inflammation grades were compared. RESULTS: There was a significant correlation between inflammation and aggressiveness scores (r = 0.39, P < 0.01). Median PSA levels in grades 1, 2 and 3 extent of inflammation were 2.4, 5.2 and 5.7 ng/ml, respectively. There was not any significant difference between these grades for PSA. Furthermore, median PSA levels in grades 1, 2 and 3 aggressiveness of inflammation were 4.4, 4.8 and 8.7 ng/ml, respectively. There was a significant difference between grades of aggressiveness of inflammation and PSA levels. CONCLUSION: High serum PSA levels may correlate with asymptomatic inflammatory prostatitis with high aggressiveness score in BPH patients without clinical prostatitis. PMID- 17111078 TI - A comparative study of different methods of orchidometry in a canine model. AB - PURPOSE: An investigation was carried out to compare the testicular volumes obtained by different methods of assessment in a series of dogs. METHOD: After obtaining ethical approval for the study, ten dogs were identified which were scheduled for castration, either at the owner's request or prior to being placed in suitable homes. A Prader orchidometer, which consists of a series of ovoid model testes inscribed with corresponding equivalent volumes, was used as an indirect method of assessing the volume of each testis. Percutaneous measurements of length and width of each testis were also carried out prior to surgery under general anaesthesia, using both a sliding and a pinch type caliper. After each testis was removed it was isolated and submerged in a beaker containing water in order to get a direct estimate of its volume by fluid displacement. Using the formula for deriving the volume of an ellipsoid from length and weight measurements, the volumes derived from the caliper measurements were estimated and compared with the orchidometer and fluid displacement values. Pearson correlation coefficients were calculated to obtain the correlations between the values produced by the four methods. RESULTS: The Prader orchidometer and the pinch type caliper both showed significant correlations, of 0.64 and 0.62 respectively, with the definitive values obtained by fluid displacement. The correlation obtained with the sliding caliper (0.4) was not significant. Despite the significant correlations, all the indirect methods consistently overestimated the real volumes obtained by fluid displacement. CONCLUSION: Estimates of testicular volumes obtained by orchidometer and pinch type caliper are useful for situations where a qualitative rather than quantitative assessment of testicular volume is required. PMID- 17111079 TI - Can serum Inhibin B and FSH levels, testicular histology and volume predict the outcome of testicular sperm extraction in patients with non-obstructive azoospermia? AB - INTRODUCTION: In our study, we evaluated the diagnostic accuracy of serum follicle stimulating hormone (FSH), Inhibin B, testicular volumes and distribution of testicular sperm extraction (TESE) outcome according to the histological diagnosis in men with non-obstructive azoospermia. MATERIALS AND METHODS: Between February 2001 and April 2002, 66 men presenting with infertility of at least 1 year were found to have non-obstructive azoospermia. Serum FSH and Inhibin B levels, testicular volumes and pathological analysis were reviewed retrospectively using medical records of these patients. RESULTS: Of 66 patients, 52 were enrolled into the study and sperm extraction was successful in 31 of 52 patients (59.6%). There was no statistically significant difference between the patients who had successful and unsuccessful TESE in terms of mean serum Inhibin B, FSH levels and testicular volumes (P>0.05). The area under ROC analysis for serum Inhibin, serum FSH and testicular volume was 0.557, 0.523 and 0.479, respectively. For Inhibin B, the best cut-off value for discriminating between successful and failed TESE at 90% sensitivity was 6.25 with a very low level of specificity (14%) and diagnostic accuracy that was 53.8. CONCLUSION: Besides the controversies about the direct marker role of serum Inhibin B in determination of spermatogenesis, it does not seem to give a clue about the prediction of sperm presence before TESE. Because of the conflicting results in the literature, the potential role of serum Inhibin B as a marker for prediction of sperm presence in testis is yet to be determined. PMID- 17111080 TI - Urological complications of congenital nephrogenic diabetes insipidus--long-term follow-up of one patient. AB - A male with a severe form of congenital nephrogenic diabetes insipidus (diuresis 10 l per day) had megaureters, megavesica, large residuum and a history of three traumatic ruptures of the megavesica and a recurrent urinary tract infection (RUTI). Hemodialysis was introduced at 41 years of age. At age of 42, he underwent a bilateral retroperitoneoscopic nephrectomy to prevent further RUTI and 8 months later transplantation of a cadaver kidney. Intermittent catheterization is necessary due to residual urine in the urinary bladder. PMID- 17111081 TI - Usefulness of an immunochromatographical assay, PSA Rapid Test as a primary screening test for prostate cancer. AB - BACKGROUND: The recent rapid increase of mass screening for prostate cancer by measuring PSA in Japan will increase the economic burden to the healthcare system. PSA Rapid Test (PRT) is a simple inexpensive test. The usefulness of PRT as a primary screening test for prostate cancer was evaluated. METHODS: When we conducted educational lectures for prostate cancer in our city, screening for prostate cancer using PRT was offered to the male participants. The results of the tests were handed to participants in writing at the end of the lectures. When the results were judged as positive, letters of referral to our institute were enclosed. RESULTS: One hundred and fourteen (18.6%) of 614 men were judged as positive by PRT. Of the 114 men with positive PRT, 73 (64%) visited our institution. Finally, 37 men underwent a transrectal prostate biopsy and a diagnosis of prostate cancer was made in 21 men (3.4% of all participants). The total costs for the PSA tests in this study were summed to be approximately $2,300, while they would be approximately $9,200 if all participants had undergone screening using the conventional quantitative method from the outset. CONCLUSION: PRT is a low-cost method to detect patients with prostate cancer. We believe the PRT is useful as an initial screening test for detecting prostate cancer and that the combination of the PRT and more precise quantitative testing would be a reasonable way to reduce the cost and achieve high detection rate. PMID- 17111082 TI - Three cycles of etoposide and cisplatin chemotherapy in clinical stage IS nonseminomatous testicular cancer. AB - PURPOSE: To assess the efficacy of three cycles of etoposide and cisplatin (EP) chemotherapy in the patients with serological disease only after orchiectomy. MATERIALS AND METHODS: Fifteen patients with nonseminomatous germ cell tumors of the testis and elevated serum tumor markers as the only evidence of persistent disease following radical orchiectomy (clinical stage IS cancer), were treated at our institution from March 1995 to February 2003. All patients received three cycles of EP chemotherapy. The toxicity was compared with that in control group consisting of 93 patients treated with three standard cycles of cisplatin, etoposide and bleomycin for good-prognosis metastatic diseases. RESULTS: The markers normalized in all patients after one (in 11 cases) or two (in 4 cases) cycles. One patient required subsequent surgery for recurrent retroperitoneal mature teratoma. All patients remained disease-free during the median follow-up period of 85 months (range: 33-128). In patients receiving EP chemotherapy less number of treatment cycles was associated with grade IV leukopenia compared to control group (p=0.04). CONCLUSIONS: A treatment program that consists of three cycles of EP caused complete disease control in all patients. The applied regimen may be considered as a therapeutic option with reduced toxicity in clinical stage IS nonseminomatous testicular cancer patients. More evidence, however, needs to be accumulated. PMID- 17111083 TI - A rare complication after circumcision: keloid of the penis. AB - Keloids are benign hyperproliferative growths of dermal collagen that usually result from excessive tissue response to skin trauma. Although benign, they may be seen as a painful and/or pruritic lesions. A 15-year-young boy was admitted to our clinic with an enlarged scar and pain on the penis for about 1 year after a routine circumcision operation. Complete surgical resection of the lesion was done and histopathological examination revealed keloid of the penis. PMID- 17111084 TI - Bilateral renal angiomyolipoma coexistent with pulmonary lymphangioleiomyomatosis and tuberous sclerosis. AB - A case of bilateral renal angiomyolipoma coexistent with pulmonary lymphangioleiomyomatosis and tuberous sclerosis was described, being in shock with massive hematuria. The left giant angiomyolipoma, filling the almost entire abdomen, was complicated with two large hump and moniliform aneurysms. This life threatening hemorrhage was successfully treated by transcatheter arterial embolization; packing mechanically detachable coils into the aneurysms and embolization of trunk of the left renal artery. Lymphangioleiomyomatosis has been suggested to be an incomplete expression of tuberous sclerosis. Although coexisting renal and pulmonary involvement in tuberous sclerosis is rare, it is important to recognize lymphangioleiomyomatosis as a pulmonary involvement of angiomyolipoma with tuberous sclerosis. PMID- 17111085 TI - Retroperitoneal lymph node dissection for high-risk stage I and stage IIA seminoma. AB - INTRODUCTION: The clinical results of radiotherapy in low-stage seminoma are excellent with negligible early morbidity. However, in a long-term follow-up various complications may occur. On the other hand, experience in nonseminomas shows that surgical morbidity has decreased markedly after invention of a nerve sparing technique. These issues served as a rationale for us to perform the primary retroperitoneal lymph node dissection (RPLND) in seminoma patients. MATERIALS AND METHODS: Fourteen pure seminoma patients (10 high-risk stage I and four with clinical stage IIA) underwent nerve-sparing RPLND from September, 1997 to December, 2002. RESULTS: Pathological evaluation revealed lymph node involvement in three out of 10 clinical stage I and in all four stage IIA cases. The patients' acceptance of the surgery was good. Minor intra- and early postoperative complications were observed in two cases. Antegrade ejaculation was preserved in all patients. No retroperitoneal or distant recurrences have been observed. All patients were free of disease with the mean follow-up period of 56 months. CONCLUSION: The excellent results and minimum morbidity of nerve-sparing RPLND together with the increased concerns on late complications of radiotherapy may turn the preference of surgery in low-stage seminoma into the subject of future discussion. PMID- 17111086 TI - Penis: an unusual site for pilonidal sinus. AB - Pilonidal sinus affects different regions of the body, gluteal cleft being the most common. This condition affecting the penis is extremely rare with only a few case reports around the world. It is prone for complications like infection, actinomycosis, abscess formation, erectile dysfunction and phimosis amongst others. We present a case of Pilonidal sinus of the penis where a pre-operative diagnosis was made and appropriate treatment in the form of circumcision prevented complications. PMID- 17111087 TI - Lymphoma of the kidney: primary or initial manifestation of rapidly progressive systemic disease? AB - Primary renal lymphoma is a controversial and rare disease. There is no agreement whether or not it is an initial manifestation of a rapidly systemic disease. Most reported cases are questionable because of incomplete staging and lack of autopsy studies. Herein, we present a 71-year-old woman, initially diagnosed with primary non-Hodgkin lymphoma of the kidney, but was lost 4 months after radical nephrectomy due to systemic disease despite aggressive chemotherapy, and suggest that, the kidney of the patient was the initial presenting site of a rapidly progressive systemic disease. PMID- 17111088 TI - Isolation, characterization and molecular evolution of a novel pearl shell lectin from a marine bivalve, Pteria penguin. AB - A novel lectin, PPL, was isolated from the mantle of penguin wing oyster (Pteria penguin) by affinity chromatography on mucin-Sepharose 4B and cation exchange chromatography on HiTrap SP. This lectin was estimated to be a 21-kDa monomer by gel filtration, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and matrix-assisted time of flight (MALDI-TOF) mass spectrometry. However, dynamic light scattering experiments revealed that a non-covalently linked dimer formed under high salt conditions (500 mM NaCl). Interestingly, PPL showed an increasing hemagglutinating activity with increasing salt concentration. The amino acid sequence of PPL was determined by direct protein sequence analysis and cDNA cloning. The 167-amino acid sequence included 24 lysine residues and had two tandemly repeated homologous domains (residues 20-78 and 107-165) with 44% internal homology. PPL showed sequence homology to L-rhamnose-binding lectins from fish eggs and a D-galactose-binding lectin from sea urchin eggs, with sequence identities in the range 37-48%. PPL agglutinated various animal erythrocytes independently of calcium ions. The minimum concentration of PPL needed to agglutinate rabbit erythrocytes was 0.5 micro g/ml, and the most effective saccharides to inhibit the hemagglutination were D-galactose, methyl-D galactopyranoside and N-acetyl-D-lactosamine. Lactose also inhibited hemagglutination, but L-rhamnose did so only weakly despite the sequence homology with trout egg L-rhamnose-binding lectins. The carbohydrate-binding specificity of PPL was further examined by frontal affinity chromatography using 37 different pyridylaminated oligosaccharides. PPL was found to have strong binding affinity for various oligosaccharides that have Galbeta1-4Glu/GlcNAc, Galbeta1 3GalNAc/GlcNAc and Galalpha 1-4Gal moieties in their structure. PPL had a high thermal stability and retained 50% of its hemagglutinating activity after incubation at 70 degrees C for 100 min. It agglutinated some Gram-negative bacteria by recognizing lipopolysaccharides. Together, these results suggest that PPL is a new member of the trout egg lectin family which participates in the self defense mechanism against bacteria and pathogens with a distinct carbohydrate binding specificity. We conclude that the trout egg lectin family proteins, in particular their carbohydrate recognition domains, have acquired diverse carbohydrate-binding specificities during molecular evolution. PMID- 17111089 TI - Molecular diversity of proteins in biological offense and defense systems. PMID- 17111090 TI - Survival and growth of two heterotrophic hydrothermal vent archaea, Pyrococcus strain GB-D and Thermococcus fumicolans, under low pH and high sulfide concentrations in combination with high temperature and pressure regimes. AB - Growth and survival of hyperthermophilic archaea in their extreme hydrothermal vent and subsurface environments are controlled by chemical and physical key parameters. This study examined the effects of elevated sulfide concentrations, temperature, and acidic pH on growth and survival of two hydrothermal vent archaea (Pyrococcus strain GB-D and Thermococcus fumicolans) under high temperature and pressure regimes. These two strains are members of the Thermococcales, a family of hyperthermophilic, heterotrophic, sulfur-reducing archaea that occur in high densities at vent sites. As actively growing cells, these two strains tolerated regimes of pH, pressure, and temperature that were in most cases not tolerated under severe substrate limitation. A moderate pH of 5.5 7 extends their survival and growth range over a wider range of sulfide concentrations, temperature and pressure, relative to lower pH conditions. T. fumicolans and Pyrococcus strain GB-D grew under very high pressures that exceeded in-situ pressures typical of hydrothermal vent depths, and included deep subsurface pressures. However, under the same conditions, but in the absence of carbon substrates and electron acceptors, survival was generally lower, and decreased rapidly when low pH stress was combined with high pressure and high temperature. PMID- 17111091 TI - Wild chimpanzee infant urine and saliva sampled noninvasively usable for DNA analyses. AB - In many genetic studies on the great apes, fecal or hair samples have been used as sources of DNA. However, feces and hairs are difficult to collect from chimpanzee infants under 3 years of age. As alternative DNA sources, we investigated the efficiency of collecting urine samples from infants compared with fecal samples, as well as the validity of the DNA extracted from urine and saliva samples of well-habituated M group chimpanzees (Pan troglodytes schweinfurthii) in the Mahale Mountains National Park, Tanzania. We collected 40 urine and 3 fecal samples from 10 infants under 3 years. Compared with feces, the urine samples were relatively easy to collect. The saliva of infants, which remained on the twigs sucked by them, was collected using cotton swabs. The average amounts of DNA extracted from the 40 urine and 6 saliva samples were 3,920 and 458 pg/mul, respectively. The rate of positive PCR was low and the allelic dropout rate was high when using less than 25 pg of template DNA in the PCR mixtures. Based on the amounts of DNA, 50% of the urine samples and 100% of the saliva samples were judged usable for accurate microsatellite genotyping. For infant chimpanzees in particular, collecting urine and saliva as an alternative to fecal and hair samples can reduce the effort invested in collection in the field. PMID- 17111092 TI - The effect of infliximab on chemokines in patients with rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is an autoimmune disease characterized by infiltration of lymphocytes, macrophages, and plasma cells into synovial membrane. The chemokines family promotes chemotactic activity in various leukocyte cell types. Chemokines thus play an essential role in the pathological formation of RA. The aim of the present study was to evaluate the influence of infliximab on serum levels of various chemokines. Twenty-four RA patients were involved in this study, which took place between March 2003 and February 2006. Infliximab was administered by intravenous infusion at a dosage of 3 mg/kg. All patients underwent general and physical examinations and routine blood and urinary analysis at the baseline, at 14 weeks, and at 30 weeks after the initial treatment. To determine whether serum and synovial fluid from RA also contained significant levels of chemokines compared with osteoarthritis patients (OA), GRO alpha, MIP-1alpha, MIP-1beta and regulated on activation normal T cell expressed and secreted (RANTES) levels of serum and synovial fluid were measured by ELISA in 20 RA patients and 20 OA patients. GRO-alpha, MIP-1beta, and RANTES levels were significantly higher in RA compared with normal volunteers, while MIP-1alpha levels showed no significant differences. The mean GRO-alpha levels in serum from RA patients treated with infliximab decreased significantly after the initial treatment. The mean RANTES and MIP-1beta levels did not change significantly after the treatment. Infliximab treatment significantly lowered the serum GRO alpha levels of RA patients. GRO-alpha is one of the crucial cytokines affected by infliximab treatment. The blocking therapy of RANTES and MIP-1beta combined with infliximab treatment may have an additional effect without competition in the TNFalpha cascade. PMID- 17111093 TI - Acute effect of cigarette smoke and nicotine on airway blood flow and airflow in healthy smokers. AB - Cigarette smoke contains irritants and vasoactive substances. We wanted to determine the effect of smoking a cigarette and of nasally or orally inhaled nicotine on airway blood flow (Q(aw)) and airflow in smokers. In ten healthy current smokers, Q(aw), FEV(1), and FEF(25-75) were measured before and at 5, 30, and 180 min after smoking a cigarette. The effects of systemic nicotine using a nicotine nasal spray and local nicotine using a nicotine inhaler were also studied. Mean (+/- SE) Q(aw) increased by 81% +/- 16% (p = 0.03) 5 min after smoking a cigarette and was no longer different from baseline at 30 and 180 min. Nicotine nasal spray and nicotine oral inhaler had no effect on Q(aw.) FEV(1) and FEF(25-75) remained unchanged after smoking a cigarette and after local or systemic nicotine administration. Smoking a cigarette is followed by a transient increase in airway blood flow but no changes in airflow. Nicotine, at the rate and dose provided by the nasal spray (systemic action) and oral inhaler (local and systemic action), does not appear to be involved in the Q(aw) change, suggesting a pharmacologic or nonspecific irritant effect of other cigarette smoke constituents. PMID- 17111094 TI - Immunogold localization of trehalose-6-phosphate synthase in leaf segments of wild-type and transgenic tobacco plants expressing the AtTPS1 gene from Arabidopsis thaliana. AB - Following the establishment of a transgenic line of tobacco (B5H) expressing the trehalose-6-phosphate synthase (TPS) gene from Arabidopsis thaliana, a preliminary immunolocalization study was conducted using leaves of adequately watered B5H and wild-type plants. Immunocytochemical staining, followed by electron microscopy showed that the enzyme could be detected in both B5H and wild type plants at two different levels. Quantification showed the signal to be two to three times higher in transgenic plants than in the wild type. This enzyme was markedly present in the vacuoles and the cell wall, and to a lesser extent in the cytosol. Moreover, a high profusion of gold particles was detected in adjacent cells and in the sieve elements. Occasional spots were also detected in chloroplasts and the nucleus, especially in the transgenic B5H line. No labeling signal was detected in mitochondria. Protein localization seems to confirm the important role of TPS in sugar metabolism and transport through the plant, which could explain its role in plant stress tolerance. Finally, it can be expected that TPS from tobacco has a relatively high similarity to the TPS of Arabidopsis thaliana. PMID- 17111095 TI - The structure and biochemistry of charophycean cell walls: I. Pectins of Penium margaritaceum. AB - Plant cell walls are essential for proper growth, development, and interaction with the environment. It is generally accepted that land plants arose from aquatic ancestors which are sister groups to the charophycean algae (i.e., Streptophyta), and study of wall evolution during this transition promises insight into structure-function relationships of wall components. In this paper, we explore wall evolutionary history by studying the incorporation of pectin polymers into cell walls of the model organism Penium margaritaceum, a simple single-cell desmid. This organism produces only a primary wall consisting of three fibrillar or fibrous layers, with the outermost stratum terminating in distinct, calcified projections. Extraction of isolated cell walls with trans-1,2 diaminocyclohexane-N,N,N',N'-tetraacetic acid yielded a homogalacturonan (HGA) that was partially methyl esterified and equivalent to that found in land plants. Other pectins common to land plants were not detected, although selected components of some of these polymers were present. Labeling with specific monoclonal antibodies raised against higher-plant HGA epitopes (e.g., JIM5, JIM7, LM7, 2F4, and PAM1) demonstrated that the wall complex and outer layer projections were composed of the HGA which was significantly calcium complexed. JIM5 and JIM7 labeling suggested that highly methyl esterified HGA was secreted into the isthmus zone of dividing cells, the site of active wall secretion. As the HGA was displaced to more polar regions, de-esterification in a non-blockwise fashion occurred. This, in turn, allowed for calcium binding and the formation of the rigid outer wall layer. The patterning of HGA deposition provides interesting insights into the complex process of pectin involvement in the development of the plant cell wall. PMID- 17111096 TI - Phospholipase C/diacylglycerol kinase-mediated signalling is required for benzothiadiazole-induced oxidative burst and hypersensitive cell death in rice suspension-cultured cells. AB - The involvement of phospholipase C/diacylglycerol kinase (PLC/DGK)-mediated signalling in oxidative burst and hypersensitive cell death was studied in rice suspension-cultured cells treated with benzothiadiazole (BTH) and infected by Xanthomonas oryza pv. oryza (Xoo), the causal agent of rice leaf blight disease. Treatment of rice suspension cells with BTH resulted in a significant oxidative burst, as indicated by accumulation of superoxide anion and H(2)O(2), and hypersensitive cell death, as determined by Evans blue staining. A peak in oxidative burst was detected 3-4 h after BTH treatment and hypersensitive cell death was observed 8 h after treatment. In addition, significant oxidative burst and hypersensitive cell death were detected in BTH-treated suspension cells, but not in untreated control cells, after Xoo infection. Scavengers and antioxidants of active oxygen species, e.g., superoxide dismutase, catalase, N-acetylcysteine, and flavone, reduced significantly the BTH-induced oxidative burst and hypersensitive cell death, indicating that oxidative burst is required for BTH induced hypersensitive cell death. Expression of the PLC/DGK pathway genes, a diacylglycerol kinase gene, OsDAGK1, and a phosphoinositide-specific phospholipase C gene, OsPI-PLC1, and a defence-related EREBP transcriptional factor gene, OsBIERF3, was activated in rice cells after BTH treatment and in the BTH-treated cells after Xoo infection. Treatment of rice cells with phosphatidic acid, a phospholipid signalling molecule, resulted in the production of oxidative burst and hypersensitive cell death. However, neomycin, a PLC inhibitor, inhibited partially but not completely the production of oxidative burst, hypersensitive cell death, and expression of OsBIERF3 and OsDAGK1 induced by BTH in rice cells. These results suggest that PLC/DGK-mediated signalling plays an important role in BTH-induced oxidative burst, hypersensitive response, and activation of defence response in rice. PMID- 17111097 TI - Role of sugars in regulating transfer cell development in cotyledons of developing Vicia faba seeds. AB - Transfer cell formation in cotyledons of developing faba bean (Vicia faba L.) seeds coincides with an abrupt change in seed apoplasm composition from one dominated by hexoses to one in which sucrose is the principal sugar. On the basis of these observations, we tested the hypothesis that sugars induce and/or sustain transfer cell development. To avoid confounding effects of in planta developmental programs, we exploited the finding that adaxial epidermal cells of cotyledons, which do not become transfer cells in planta, can be induced to form functional transfer cells when cotyledons are cultured on an agar medium. Growth rates of cotyledons cultured on hexose or sucrose media were used to inform choice of sugar concentrations. The same proportion of adaxial epidermal cells of excised cotyledons were induced to form wall ingrowths independent of sugar species and concentration supplied. In all cases, induction of wall ingrowths coincided with a marked increase in the intracellular sucrose-to-hexose ratio. In contrast, further progression of wall ingrowth deposition was correlated positively with intracellular sucrose concentrations that varied depending upon external sugar species and supply. Sucrose symporter induction and subsequent maintenance behaved identically to wall ingrowth formation in response to an external supply of hexoses or sucrose. However, in contrast to wall ingrowth formation, induction of sucrose symporter activity was delayed. We discuss the possibility of intracellular sugars functioning both as signals and substrates that induce and control subsequent development of transfer cells. PMID- 17111098 TI - Symbiotic alga Chlorella vulgaris of the ciliate Paramecium bursaria shows temporary resistance to host lysosomal enzymes during the early infection process. AB - Paramecium bursaria free of symbiotic Chlorella species can be experimentally reinfected with algae isolated from algae-bearing cells by ingestion into digestive vacuoles. Isolated symbiotic algae were cloned, mixed with the algae free P. bursaria at 25 +/- 1 degrees C for 1.5 min, washed and chased, with or without fixation 3 h after mixing. Though genetically identical, a few of the algae were not digested but coexisted with the digested ones in the same vacuole after lysosomal fusion. Light microscopy showed that algal fate did not depend on cell cycle stage or location in the vacuole. Electron microscopy showed that the nondigested algae were not protected by a perialgal vacuole membrane in the digestive vacuole. Moreover, this phenomenon was also observed in the presence of cycloheximide and puromycin, which are known to inhibit algal and host protein synthesis, respectively. These observations suggest that a few algae can acquire temporary resistance to host lysosomal enzymes in order to establish endosymbiosis without algal protein synthesis. PMID- 17111099 TI - Phosphorylation of H2AX histones in response to double-strand breaks and induction of premature chromatin condensation in hydroxyurea-treated root meristem cells of Raphanus sativus, Vicia faba, and Allium porrum. AB - Histone H2A variant H2AX is rapidly phosphorylated on the induction of DNA double strand breaks by ionizing radiation and hydroxyurea-mediated replication arrest, resulting in the formation of gamma-H2AX foci along megabase chromatin domains nearby the sites of incurred DNA damage. In an attempt to establish a relationship between species-specific nuclear architecture and H2AX phosphorylation in S/G(2) phase-arrested root meristem cells, immunocytochemical comparisons using an antibody raised against human gamma-H2AX were made among three plants differing with respect to DNA contents: Allium porrum, representing a reticulate type of DNA package, Vicia faba, having semireticulate cell nuclei, and Raphanus sativus, characterised by a chromocentric type of chromatin. Another approach was aimed at determining possible correlations between the extent of hydroxyurea-induced phosphorylation of H2AX histones and the quantities of root meristem cells induced by caffeine to enter aberrant mitotic division (premature chromosome condensation). It was concluded that the higher-order structure of chromatin may contribute to the accessibility of molecular factors engaged in the recognition and repair of genetic lesions. Consequently, in contrast to A. porrum and V. faba, a diffuse chromatin in chromocentric cell nuclei of R. sativus may become more vulnerable both to generate DNA double-strand breaks and to recruit molecular elements needed to arrange the cell cycle checkpoint functions, and thus, more resistant to factors which allow the cells to enter premature chromosome condensation spontaneously. On the other hand, however, caffeine mediated overriding of the S-M checkpoint control system resulted in the typical appearance of premature chromosome condensation, irrespective of the genomic content of DNA. PMID- 17111100 TI - Differential responses to anxiogenic drugs in a mouse model of panic disorder as revealed by Fos immunocytochemistry in specific areas of the fear circuitry. AB - Sensitivity to pharmacological challenges has been reported in patients with panic disorder. We have previously validated transgenic mice overexpressing the neurotrophin-3 (NT-3) receptor, TrkC (TgNTRK3), as an engineered murine model of panic disorder. We could determine that TgNTRK3 mice presented increased cellularity in brain regions, such as the locus ceruleus, that are important neural substrates for the expression of anxiety in severe anxiety states. Here, we investigated the sensitivity to induce anxiety and panic-related symptoms by sodium lactate and the effects of various drugs (the alpha2-adrenoceptor antagonist, yohimbine and the adenosine antagonist, caffeine), in TgNTRK3 mice. We found enhanced panicogenic sensitivity to sodium lactate and an increased intensity and a differential pattern of Fos expression after the administration of yohimbine or caffeine in TgNTRK3. Our findings validate the relevance of the NT-3/TrkC system to pathological anxiety and raise the possibility that a specific set of fear-related pathways involved in the processing of anxiety related information may be differentially activated in panic disorder. PMID- 17111101 TI - 1-(N-chloroacetylamino)-alkylphosphonic acids - synthetic precursors of phosphonopeptides. AB - General procedures of N-chloroacetylation of the representative 1 aminoalkylphosphonic acids (Gly(P), Ala(P), Val(P), Pgly(P) and Phe(P)) are described. These 1-(N-chloroacetylamino)-alkylphosphonic acids were converted into the corresponding glycylphosphonodipeptides (Gly-AA(P)) and/or related N alkylglycylphosphonodipeptides (Me(n)Gly-AA(P)) in the course of ammonolysis/aminolysis. Physico-chemical properties of synthesized 1-(N chloroacetylamino)-alkylphosphonic acids and phosphonodipeptides are characterized. PMID- 17111102 TI - Laryngeal pemphigus without skin manifestations and review of the literature. AB - Pemphigus is an uncommon chronic disease with dermatologic and mucosal manifestations. Primary laryngeal involvement without skin lesions is extremely rare. The present paper describes a 72-year old man who presented with a 2-month history of hoarseness, haemoptisis and dysphagia. Clinical examination revealed an erythematous oral mucosa without ulcerations. Indirect laryngoscopy revealed supraglottic ulcerations mainly in the laryngeal surface of the epiglottis and in the right arytenoid. The lesions had characteristic gray color membranes. The patient underwent microlaryngoscopy under general anesthesia and biopsies were taken for histology that revealed inflammatory and granular lesions with necrosis. The diagnosis of pemphigus was based on immunohistopathology and the clinical examination. The patient underwent intravenous treatment with high doses of corticosteroids (prezolon 75 mg/24 h) for 10 days and gradually the dose was reduced to 10 mg/24 h. The patient had a very good response to the treatment and after a week approximately 80% of the lesions disappeared. However, the dose of 10 mg prednisolone per day was sustained for 3 months because any attempt of prednisolone discontinuation was related with reappearance of the clinical symptoms. After 3 months, finally the treatment was discontinued without problems. Now, 15 months later, the patient is well and without symptoms. He is under long-term follow-up. ENT surgeons should be aware of pemphigus as primary laryngeal manifestation in order to investigate and manage patients accordingly. PMID- 17111103 TI - Determination of 90Sr in contaminated environmental samples by tuneable bandpass dynamic reaction cell ICP-MS. AB - A rapid method for the extraction and determination of 90Sr in natural water, plant and sediment samples was developed using extraction chromatography and dynamic reaction cell ICP-MS, with O2 as a reaction gas. While isobaric interference from the stable isotope 90Zr was efficiently removed by this method, interferences produced from in-cell reactions with Fe+ and Ni+ required suppression by tuneable bandpass, and in sediments, additional chromatographic separation. Method detection limits were 0.1 pg g-1 (0.5 Bq g-1), 0.04 pg g-1(0.2 Bq g-1), and 3 pg L-1 (5 Bq L-1) for sediments, plant and water samples, respectively, and 90Sr concentrations determined by ICP-MS were in good agreement with activities determined by Cerenkov counting and with certified reference values. While mass spectrometric determination does not rival detection limits achievable by radiometric counting, radiometric determination of 90Sr, a pure beta-emitter, is hindered by long analysis times (several weeks); the comparatively fast analysis achieved via ICP-MS enables same-day preparation and analysis of samples, making this an important technique for the environmental monitoring of areas contaminated by radioactivity. PMID- 17111104 TI - Neurochemical challenge. PMID- 17111106 TI - Publication of chemical research: do we need ethical standards? PMID- 17111107 TI - William John Sibbald: in memoriam. PMID- 17111108 TI - Research, a never-ending jigsaw puzzle. Two new series in Intensive Care Medicine. PMID- 17111109 TI - The Nuremberg doctors' trial: the 60th anniversary. PMID- 17111110 TI - Positive end-expiratory pressure and pressure support in peripheral airways obstruction : work of breathing in intubated children. AB - OBJECTIVES: Children with peripheral airways obstruction suffer the negative effects of intrinsic positive end-expiratory pressure: increased work of breathing and difficulty triggering assisted ventilatory support. We examined whether external positive end-expiratory pressure to offset intrinsic positive end-expiratory pressure decreases work of breathing in children with peripheral airways obstruction. The change in work of breathing with incremental pressure support was also tested. DESIGN AND SETTING: Prospective clinical trial in a pediatric intensive care unit. PATIENTS: Eleven mechanically ventilated, spontaneously breathing children with peripheral airways obstruction. INTERVENTIONS: Work of breathing (using pressure-rate product as a surrogate) was measured in three tiers: (a) Increasing pressure support over zero end-expiratory pressure. (b) Increasing applied positive end-expiratory pressure and fixed pressure support. The level of applied positive end-expiratory pressure at which pressure-rate product was least determined the compensatory positive end expiratory pressure. (c) Increasing pressure support over compensatory (fixed) positive end-expiratory pressure. MEASUREMENTS AND RESULTS: Increases in pressure support alone decreased pressure-rate product from mean 724+/-311 to 403+/-192 cmH2O/min. Applied positive end-expiratory pressure alone decreased pressure-rate product from mean 608+/-301 to 250+/-169 cmH2O/min. The lowest pressure-rate product (136+/-128 cmH2O/min) was achieved using compensatory positive end expiratory pressure (12+/-4 cmH2O) with pressure support 16 cmH2O. CONCLUSIONS: For children with peripheral airways obstruction who require assisted ventilation, work of breathing during spontaneous breaths is decreased by the application of either compensatory positive end-expiratory pressure or pressure support. PMID- 17111111 TI - Tocopherol biosynthesis is enhanced in photomixotrophic sunflower cell cultures. AB - Alpha-tocopherol is the most biologically active component of vitamin E and is synthesized only by photosynthetic organisms. Two heterotrophic cell lines of sunflower (Helianthus annuus L.) of differing alpha-tocopherol biosynthetic capability, three-fold higher in the high synthesizing cell line, HT, than in the low synthesizing one, LT, were previously identified. To investigate the relationship between alpha-tocopherol biosynthesis and photomixotrophic culture conditions, a new photomixotrophic sunflower cell line HS3 was established by selecting HT cells able to grow in the presence of a ten-fold reduced sucrose concentration in the culture medium. The photosynthetic properties of HS3 cells were characterized in comparison with HT and LT cells, revealing an increase in chlorophyll content, chloroplast number, and level of the photosynthesis related enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Furthermore, an enhanced expression of the gene encoding for the tocopherol biosynthetic enzyme geranyl-geranylpyrophosphate synthase (GGPPS) was observed in HS3 cells. HS3 cells also revealed a 25% and a more than three-fold higher tocopherol level than HT and LT, respectively, indicating a positive correlation between alpha tocopherol biosynthesis of sunflower cell cultures and their photosynthetic properties. These findings can be useful for improving the tocopherol yields of the sunflower in vitro production system. PMID- 17111112 TI - Applications of DL-buthionine-[S,R]-sulfoximine deplete cellular glutathione and improve white spruce (Picea glauca) somatic embryo development. AB - In white spruce (Picea glauca), an improvement of somatic embryo yield and quality can be achieved by applications of DL: -buthionine-[S,R]-sulfoximine (BSO), which inhibits the biosynthesis of reduced glutathione (GSH), thereby switching the total glutathione pool towards its oxidized form (GSSG). Applications of BSO almost tripled the embryogenic output of two cell lines by increasing the number of embryos produced by 100 mg(-1) tissue from 65 to 154 in the (E)WS1 line and from 59 to 130 in the (E)WS2 line. This increase in embryo number was ascribed to a higher production of morphologically normal embryos with four or more cotyledons (group A embryos), at the expense of group B embryos, characterized by fewer cotyledons. The quality of the embryos produced, estimated by their post-embryonic performance, was also different between treatments. In both cell lines applications of BSO in the maturation medium increased the conversion frequency, i.e. root and shoot emergence, of group A embryos while it enhanced root emergence in group B embryos. Compared to their control counterparts, BSO-treated embryos had normal shoot apical meristems as in their zygotic counterparts. Such meristems were characterized by large apical cells and vacuolated sub-apical cells. They also lacked intercellular spaces, which were present in the apical poles of control embryos where they contributed to cell cell separation and meristem degradation. Furthermore, storage product accumulation was also improved in the presence of BSO, with protein bodies prevailing over starch. These data show that an oxidized glutathione environment is beneficial for spruce embryo production in vitro. PMID- 17111113 TI - Compact shoot and leafy head 1, a mutation affects leaf initiation and developmental transition in rice (Oryza sativa L). AB - The shoot apical meristem (SAM) produces lateral organs in a regular spacing (phyllotaxy) and at a regular interval (phyllochron) during the vegetative phase. In a Dissociation (Ds) insertion rice population, we identified a mutant, compact shoot and leafy head 1 (csl1), which produced massive number of leaves (~70) during the vegetative phase. In csl1, the transition from the vegetative to the reproductive phase was delayed by about 2 months under long-day conditions. With a reduced leaf size and severe dwarfism, csl1 failed to produce a normal panicle after the transition to reproductive growth. Instead, it produced a leafy panicle, in which all primary rachis-branches were converted to vegetative shoots. Phenotypically csl1 resembled pla mutants in short plastochron but was more severe in the conversion of the reproductive organs to vegetative organs. In addition, neither the expression nor the coding region of PLA1 or PLA2 was affected in csl1. csl1 is most likely a dominant mutation because no mutant segregant was observed in progeny of 67 siblings of the csl1 mutant. CSL1 may represent a novel gene, which functions downstream of PLA1 and/or PLA2, or alternatively functions in a separate pathway, involved in the regulation of leaf initiation and developmental transition via plant hormones or other mobile signals. PMID- 17111114 TI - Children's coping with in vivo peer rejection: an experimental investigation. AB - We examined children's behavioral coping in response to an in vivo peer rejection manipulation. Participants (N=186) ranging between 10 and 13 years of age, played a computer game based on the television show Survivor and were randomized to either peer rejection (i.e., being voted out of the game) or non-rejection control. During a five-min. post-feedback waiting period children's use of several behavioral coping strategies was assessed. Rejection elicited a marked shift toward more negative affect, but higher levels of perceived social competence attenuated the negative mood shift. Children higher in depressive symptoms were more likely to engage in passive and avoidant coping behavior. Types of coping were largely unaffected by gender and perceived social competence. Implications are discussed. PMID- 17111115 TI - Investigation for cystic fibrosis in infants with jejunoileal atresia in the Netherlands: a 35-year experience with 114 cases. PMID- 17111116 TI - Your diagnosis? Umbilical cord tumor. PMID- 17111117 TI - Automatic user customization for improving the performance of a self-paced brain interface system. AB - Customizing the parameter values of brain interface (BI) systems by a human expert has the advantage of being fast and computationally efficient. However, as the number of users and EEG channels grows, this process becomes increasingly time consuming and exhausting. Manual customization also introduces inaccuracies in the estimation of the parameter values. In this paper, the performance of a self-paced BI system whose design parameter values were automatically user customized using a genetic algorithm (GA) is studied. The GA automatically estimates the shapes of movement-related potentials (MRPs), whose features are then extracted to drive the BI. Offline analysis of the data of eight subjects revealed that automatic user customization improved the true positive (TP) rate of the system by an average of 6.68% over that whose customization was carried out by a human expert, i.e., by visually inspecting the MRP templates. On average, the best improvement in the TP rate (an average of 9.82%) was achieved for four individuals with spinal cord injury. In this case, the visual estimation of the parameter values of the MRP templates was very difficult because of the highly noisy nature of the EEG signals. For four able-bodied subjects, for which the MRP templates were less noisy, the automatic user customization led to an average improvement of 3.58% in the TP rate. The results also show that the inter subject variability of the TP rate is also reduced compared to the case when user customization is carried out by a human expert. These findings provide some primary evidence that automatic user customization leads to beneficial results in the design of a self-paced BI for individuals with spinal cord injury. PMID- 17111119 TI - Weekly administration of topotecan-paclitaxel as second-line treatment in ovarian cancer. AB - PURPOSE: To investigate the weekly administration of topotecan combined with paclitaxel in pretreated advanced ovarian cancer patients; our objectives were to determine efficacy, toxicity and survival. METHODS: The chemotherapy agents, topotecan and paclitaxel were administered on a weekly basis for 3 consecutive weeks, every 28 days. The plan was to give three courses (each course included three once-weekly infusions). The dose of topotecan was 1.75 mg/m(2) and of paclitaxel 70 mg/m(2). RESULTS: From January 2004 until January 2006, 45 patients were enrolled in this multicenter trial; 44 patients were evaluable for response and toxicity. The median age was 60 years old (range 39-82 years) and performance status was 0-2. Thirty-nine patients were in stage III and 5 in stage IV. All patients had been pretreated with carboplatin or cisplatin in combination with paclitaxel. Complete and partial responses were seen in 39% of the patients, stable disease in 43% and progressive disease in 18%; median survival time was 9 months, range 2-24+ months, (95% CI: 7.9-10.2). There was a notable absence of grade 3 toxicity except for neutropenia in 11% of the patients. CONCLUSION: The combination of topotecan and paclitaxel administered on a weekly basis is a well tolerated chemotherapy schedule. The response rate of 39% is quite high for patients with pretreated ovarian cancer. PMID- 17111118 TI - Heart rate variability: a review. AB - Heart rate variability (HRV) is a reliable reflection of the many physiological factors modulating the normal rhythm of the heart. In fact, they provide a powerful means of observing the interplay between the sympathetic and parasympathetic nervous systems. It shows that the structure generating the signal is not only simply linear, but also involves nonlinear contributions. Heart rate (HR) is a nonstationary signal; its variation may contain indicators of current disease, or warnings about impending cardiac diseases. The indicators may be present at all times or may occur at random-during certain intervals of the day. It is strenuous and time consuming to study and pinpoint abnormalities in voluminous data collected over several hours. Hence, HR variation analysis (instantaneous HR against time axis) has become a popular noninvasive tool for assessing the activities of the autonomic nervous system. Computer based analytical tools for in-depth study of data over daylong intervals can be very useful in diagnostics. Therefore, the HRV signal parameters, extracted and analyzed using computers, are highly useful in diagnostics. In this paper, we have discussed the various applications of HRV and different linear, frequency domain, wavelet domain, nonlinear techniques used for the analysis of the HRV. PMID- 17111120 TI - Biweekly oxaliplatin plus 1-day infusional fluorouracil/leucovorin followed by metronomic chemotherapy with tegafur/uracil in pretreated metastatic colorectal cancer. AB - PURPOSE: Metronomic chemotherapy, at a minimally toxic dose and with a frequent schedule, is a potentially novel approach to the control of advanced cancer disease via a different mechanism from maximum tolerable doses chemotherapy. Taking advantage of the potential effectiveness of metronomic therapy, tegafur/uracil (UFT) was incorporated into an oxaliplation/infusioanl fluouracil (5-FU)/leucovorin (LV) protocol in this study. The primary endpoints were response rate, time to progression (TTP) and safety profile in 5-FU-pretreated metastatic colorectal cancers (CRCs). PATIENTS AND METHODS: Twenty-eight patients with metastatic CRCs resistant or refractory to 5-FU/LV were enrolled. Chemotherapy was administrated every 2 weeks sequentially with 2-h infusion of oxaliplatin (85 mg/m2) and LV (200 mg/m2), intravenous bolus 5-FU (400 mg/m2), 22 h infusion of 5-FU (600 mg/m2) on day 1 and then followed by 10-day daily oral UFT (200 mg/m2)/LV (30 mg/m2). RESULTS: Partial response was seen in ten (35.7%) patients. The median TTP was 5.2 (95% CI: 4.16-6.31) months and the median overall survival was 13.4 (95% CI: 6.39-20.5) months. No grade 3 toxicities above 5% according to National Cancer Institute-Common Toxicity Criteria (NCI-CTC) occurred except sensory neuropathy (10.7%). No grade 4 toxicity, treatment related mortality or hand-foot syndrome was found. CONCLUSIONS: This study protocol with favorable toxicity profile is thus promisingly effective against 5 FU-pretreated metastatic CRCs. Given the present experience, an evaluation of the regimen as front-line treatment of metastatic CRC is planned. PMID- 17111121 TI - Comparison of the proliferative activity in gingival epithelium after surgical treatments of intrabony defects with bioactive glass and bioabsorbable membrane. AB - Guided tissue regeneration is based on preventing the more rapidly proliferating epithelium from growing into the periodontal defect after surgical procedures incorporating barrier membranes. The aim of this study was to compare the proliferative activity of gingival epithelium using proliferating cell nuclear antigen (PCNA) as a marker of cell proliferation after surgical treatments with bioactive glass graft material and bioabsorbable membrane. Using split mouth design, 20 intrabony defects were randomly assigned treatments with bioactive glass (BG group) or bioabsorbable membrane (BM group). Gingival biopsies were taken at preoperative and postoperative 12 weeks. After histological processing, the number of the inflammatory cells was measured in hematoxylin and eosin stained sections; PCNA expression was determined in immunohistochemically-stained sections. At postoperative 12 weeks, the number of the inflammatory cells was significantly decreased (p < 0.01), PCNA expression was significantly increased (p < 0.001) in both treatment groups compared to baseline data. There was no significant difference in PCNA expression between baseline values of two groups (p > 0.05), while at postoperative 12 weeks, increase in BG group was significantly greater than that in BM group (p < 0.001). These results suggest that epithelial cell proliferation is more prominent after treatment of intrabony defects with bioactive glass compared to the treatment with bioabsorbable membrane. PMID- 17111122 TI - High-molecular tenascin-C as an indicator of atypical cells in oral brush biopsies. AB - Tumour-invasion like wound healing is characterised by the formation of an extracellular matrix with a high tenascin-C content. The tenascin-C molecule undergoes alternative splicing. Analysis using antibody BC2 indicates that especially the high-molecular tenascin-C (hm tn-C) variants are typically tumour associated, while distribution in normal tissue is restrictive. This study investigated whether hm tn-C is a suitable indicator of atypical cells with invasive potential in oral brush biopsies. One hundred fifty nine consecutive oral brush biopsies with histopathological diagnoses were analysed for the identification of atypical cells. A standardised haematoxylin and eosin staining plus standardised immunocytochemistry using the monoclonal anti-hm tn-C antibody was performed. The bound hm tn-C antibodies were detected with the streptavidine/alkaline phosphatase technique in the autostainer. Conventional cytology produced four false-positives when identifying atypical cells in brush biopsies of inflammatory/benign hyperproliferative mucosa (specificity 96%), while 10 in 52 carcinomas and three of eight recurrences were not identified (sensitivity 78%). Ten of these 13 non-identified tumours could be marked when adding the hm tn-C assay (increasing specificity to 99%). Combining the two assays also reduced the false-positive outcomes from four to one (increasing sensitivity to 95%). The positive and negative predictive values were 92 and 88% for conventional cytology vs 98 and 97% for the dual assay. (1) A 95%-sensitivity proves hm tn-C assisted conventional cytology to be a suitable means of identifying atypical cells in oral brush biopsies. (2) The positive (98%) and negative (97%) predictive values obtained approximate hm tn-C assisted conventional cytology to laminin-5 (100/97%). PMID- 17111123 TI - Cystic endosalpingiosis of the appendix. PMID- 17111124 TI - High-risk human papillomavirus type 16 E7 oncogene associates with Cdc25A over expression in oral squamous cell carcinoma. AB - Cells expressing high-risk human papillomavirus (HPV) E7 protein display impaired checkpoint control after DNA damage and exhibit elevated rates of mutagenesis. Repression of HPV E7 expression results in the subsequent accumulation of hypophosphorylated retinoblastoma protein and repression of the Cdc25A genes. No study has been conducted to elucidate the role of Cdc25A in the development and progression of human oral carcinomas. To confirm Cdc25A protein expression together with HPV, immunohistochemistry, Western blotting, polymerase chain reaction (PCR), and reverse transcriptase (RT)-PCR were performed using various histological subtypes of oral carcinomas. Cdc25A protein was localized predominantly in the cell nuclei in carcinomas, and high expression was found in 54% of primary tumors. HPV-16 E7 was not found in non-neoplastic oral tissues, whereas it was observed in eight (36%) of 22 oral carcinomas. We found a significant correlation between Cdc25A over-expression and HPV-16 E7 positive carcinomas. There was a strong positive correlation between Cdc25A over expression and tumor size and TNM stage. This study suggests that Cdc25A is likely to be an important mediator in the progression of oral tumors, and HPV-16 E7 may be a sensitive indicator of the involvement of viral oncogenes in oral carcinogenesis. PMID- 17111125 TI - Late recurrence of Wilms' tumour with exclusive skeletal muscle phenotype 23 years after primary diagnosis. AB - A late recurrence of Wilms' tumour (WT) 23 years after the primary diagnosis is described. The primary tumour occurred in a 10-month-old girl and showed various degrees of differentiation, including skeletal muscle phenotype. A postoperative chemotherapy was performed. Twenty-three years after the surgery, the tumour relapsed: the lesion was exclusively composed of mature skeletal muscle elements (diffuse and intense desmin reactivity) derived from the primary tumour as confirmed by WT1 immunoreactivity. Chemotherapy and radiotherapy have been reported previously to ablate the immature components of WT; especially, chemotherapy can modify the histological type, reducing the immature elements while leaving mature cells unaffected. We can hypothesise that both morphological and molecular features of the tumour as well as the effect of therapy can influence a tumour relapse in WT. The latter results in a high degree of differentiation and a long disease-free interval after the first diagnosis. PMID- 17111126 TI - A case of acquired renal cystic disease (ACDK) with oncocytosis, a dominant nodule (oncocytoma), multiple adenomas and a microscopic papillary renal cell carcinoma associated with crescentic glomerulonephritis. PMID- 17111127 TI - Crystal-storing histiocytosis associated with MALT lymphoma of the ocular adnexa: a case report with review of literature. AB - We report a case of crystal-storing histiocytosis (CSH) associated with mucosa associated lymphoid tissue (MALT) lymphoma of the ocular adnexa. The patient was an 81-year-old woman who presented with a 5-month history of a slowly enlarging tumor on her left lower eyelid. The tumor was 2 cm in the largest diameter, involving both inferior oblique and inferior rectus muscles. Histological examination revealed that the tumor was composed predominantly of sheets of spindle-shaped cells resembling striated muscle cells, and scattered aggregates of atypical lymphoid cells at the periphery of the tumor, showing prominent plasmacytoid differentiation. Immunohistochemical and ultrastructural analyses demonstrated that the spindle-shaped cells were CD68-positive histiocytes containing abundant rod-like and/or rectangular crystals in their cytoplasm, consistent with the diagnosis of CSH. The scattered aggregates of atypical lymphoid cells were diagnosed as MALT lymphoma based upon their immunophenotype, featuring diffusely positive staining for CD20, but negative for CD3, CD5, and CD10, and monotypic expression of IgM-kappa in cells with plasmacytoid differentiation. Although CSH is a well-recognized manifestation in lymphoproliferative disorders in the literature, CSH complicated by MALT lymphoma has only very rarely been reported. Given the rarity of this, difficulties in diagnosis may arise especially in cases where histiocytic proliferation overwhelms the underlying lymphoproliferative diseases. PMID- 17111128 TI - So-called infantile haemangiopericytoma of the kidney. PMID- 17111129 TI - CD24 expression in urothelial carcinoma of the upper urinary tract correlates with tumour progression. AB - Expression of the mucin-like adhesion molecule CD24 has been implicated in the progression of several types of cancer and has been identified as new prognostic factor. We evaluated CD24 expression in 268 consecutive cases of upper urinary tract urothelial carcinoma with respect to associations with tumour stage, grade, angioinvasion and infiltrative growth pattern using a tissue microarray technique and correlated data with patient outcome. CD24 expression was demonstrated in 161/259 (62%) evaluable tumours and was associated with high tumour stage [77/139 (55%) pTa/pT1 vs 84/120 (70%) pT2-pT4; P=0.02] and high tumour grade [68/139 (49%) low vs 93/120 (78%) high grade; P<0.001] as well as presence of angioinvasion (P=0.002) and infiltrative pattern of invasion (P=0.007). Patients with CD24-positive tumours tended to have a higher risk of disease progression (P=0.065). Multivariate analysis, however, proved pT stage >1 [P<0.001, risk ratio (RR)=5.87, 95% confidence interval (CI)=2.88-11.95] and high tumour grade (P<0.001, RR=3.30, 95% CI 1.75-6.22) as only independent predictors of metastatic disease. In conclusion, CD24 expression in upper urinary tract urothelial cancer is associated with advanced tumour stage and high tumour grade as well as histopathological features indicative of aggressive tumour behaviour, but it lacks independent impact on patient outcome. PMID- 17111130 TI - Small hepatic vein involvement in IgG multiple myeloma: a very unusual pattern of liver infiltration. PMID- 17111131 TI - Inulinase production by a marine yeast Pichia guilliermondii and inulin hydrolysis by the crude inulinase. AB - Marine yeast strain 1, isolated from the surface of a marine alga, was found to secrete a large amount of inulinase into the medium. This marine yeast was identified as a strain of Pichia guilliermondii according to the results of routine yeast identification and molecular methods. The crude inulinase produced by this marine yeast worked optimally at pH 6.0 and 60 degrees C. The optimal medium for inulinase production was seawater containing 4.0% (w/v) inulin and 0.5% (w/v) yeast extract, while the optimal cultivation conditions for inulinase production were pH 8.0, 28 degrees C and 170 rpm. Under the optimal conditions, over 60 U ml(-1) of inulinase activity was produced within 48 h of fermentation in shake flasks. A large amount of monosaccharides and a trace amount of oligosaccharides were detected after the hydrolysis, indicating that the crude inulinase had a high exoinulinase activity. PMID- 17111132 TI - Characterization of the gene transfer process mediated by histidine-rich peptides. AB - Peptides have an interesting potential as gene transfer agents not only because they are biodegradable but also due to their reduced size and the facility of product characterization and large-scale production. Recently, we have shown that the cationic amphipathic 26-mer peptide LAH4 possesses high DNA delivery capacities. To better understand its way of functioning and to further improve its efficiency, we synthesized and tested various LAH4 derivatives. The evaluation of their DNA delivery capacity, as well as their transfection efficiency, makes accessible valuable information about structure-function correlations. In particular, our results indicate the threshold peptide concentration for endosomal escape and conformational preferences for the peptide. Furthermore, the results indicate that a fine balance of peptide-DNA interactions is responsible for the high transfection activity of LAH4. Taken together, the data also suggest that the peptides efficiently transport DNA into the cytoplasm and that the creation of more potent transfection compounds probably needs improvement of other steps during the process. PMID- 17111133 TI - Production of GDP-L-fucose, L-fucose donor for fucosyloligosaccharide synthesis, in recombinant Escherichia coli. AB - A recombinant Escherichia coli strain was developed to produce guanosine 5' diphosphate (GDP)-L-fucose, donor of L-fucose, which is an essential substrate for the synthesis of fucosyloligosaccharides. GDP-D: -mannose-4, 6-dehydratase (GMD) and GDP-4-keto-6-deoxymannose 3, 5-epimerase 4-reductase (WcaG), the two crucial enzymes for the de novo GDP-L-fucose biosynthesis, were overexpressed in recombinant E. coli by constructing inducible overexpression vectors. Optimum expression conditions for GMD and WcaG in recombinant E. coli BL21(DE3) were 25 degrees C and 0.1 mM isopropyl-beta-D-thioglucopyranoside. Maximum GDP-L-fucose concentration of 38.9 +/- 0.6 mg l(-1) was obtained in a glucose-limited fed batch cultivation, and it was enhanced further by co-expression of NADPH regenerating glucose-6-phosphate dehydrogenase encoded by the zwf gene to achieve 55.2 +/- 0.5 mg l(-1) GDP-L-fucose under the same cultivation condition. PMID- 17111134 TI - Fabrication and application of carbohydrate microarray for analyzing human serum antibody-carbohydrate interaction. AB - We introduced a strategy for preparing a carbohydrate microarray and demonstrated its utility for characterizing carbohydrate binding and activities. We isolated the lipopolysaccharide (LPS) components from different bacteria and explored the possibility of immobilizing these glycoconjugates on a high-binding polystyrene plate. Carbohydrate-specific combination was examined by observing the binding of the blood group B analogic LPS O-polysaccharide from Escherichia coli on the high binding polystyrene plate and anti-B from a broad spectra antibody of human blood serum. Strong binding of antibodies was screened, as it was evident that relative response value is two times higher than control. The hybridization results indicated that this method is a reliable technique for the detection of human intestinal bacteria and is expected to be applied in diagnostics and seroepidemiology. PMID- 17111135 TI - Efficient induction of formate hydrogen lyase of aerobically grown Escherichia coli in a three-step biohydrogen production process. AB - A three-step biohydrogen production process characterized by efficient anaerobic induction of the formate hydrogen lyase (FHL) of aerobically grown Escherichia coli was established. Using E. coli strain SR13 (fhlA (++), DeltahycA) at a cell density of 8.2 g/l medium in this process, a specific hydrogen productivity (28.0 +/- 5.0 mmol h(-1) g(-1) dry cell) of one order of magnitude lower than we previously reported was realized after 8 h of anaerobic incubation. The reduced productivity was attributed partly to the inhibitory effects of accumulated metabolites on FHL induction. To avoid this inhibition, strain SR14 (SR13 DeltaldhA DeltafrdBC) was constructed and used to the effect that specific hydrogen productivity increased 1.3-fold to 37.4 +/- 6.9 mmol h(-1) g(-1). Furthermore, a maximum hydrogen production rate of 144.2 mmol h(-1) g(-1) was realized when a metabolite excretion system that achieved a dilution rate of 2.0 h(-1) was implemented. These results demonstrate that by avoiding anaerobic cultivation altogether, more economical harvesting of hydrogen-producing cells for use in our biohydrogen process was made possible. PMID- 17111136 TI - Applications of whole-cell bacterial sensors in biotechnology and environmental science. AB - Biosensors have major advantages over chemical or physical analyses with regard to specificity, sensitivity, and portability. Recently, many types of whole-cell bacterial biosensors have been developed using recombinant DNA technology. The bacteria are genetically engineered to respond to the presence of chemicals or physiological stresses by synthesizing a reporter protein, such as luciferase, beta-galactosidase, or green fluorescent protein. In addition to an overview of conventional biosensors, this minireview discusses a novel type of biosensor using a photosynthetic bacterium as the sensor strain and the crtA gene, which is responsible for carotenoid synthesis, as the reporter. Since bacteria possess a wide variety of stress-response mechanisms, including antioxidation, heat-shock responses, nutrient-starvation, and membrane-damage responses, DNA response elements for several stress-response proteins can be fused with various reporter genes to construct a versatile set of bacterial biosensors for a variety of analytes. Portable biosensors for on-site monitoring have been developed using a freeze-dried biosensing strain, and cell array biosensors have been designed for high-throughput analysis. Moreover, in the future, the use of single-cell biosensors will permit detailed analyses of samples. Signals from such sensors could be detected with digital imaging, epifluorescence microscopy, and/or flow cytometry. PMID- 17111137 TI - The role of lipids in the biogenesis of integral membrane proteins. AB - Most integral membrane proteins are cotranslationally inserted into the lipid bilayer. In prokaryotes, membrane insertion of the nascent chain takes place at the plasma membrane, whereas in eukaryotes insertion takes place into the endoplasmatic reticulum. In both kingdoms of life, however, the same membrane that acquaints the newly born membrane protein also synthesizes the bilayer lipids and thus ensures the balanced growth of the membrane as a whole. Recent evidence indicates that the lipid composition of the host membrane can determine the fate of the newborn membrane protein, as it can affect (1) the efficiency of translocation, (2) the topology of the resulting membrane protein, (3) its stability, (4) its assembly into oligomeric complexes, (5) its transport and sorting along the secretory pathway, and (6) its enzymatic activity. The lipid composition of the membrane thus can affect the biogenesis and function of integral membrane proteins at multiple steps along its biogenetic pathway. While understanding this interdependence between bilayer lipids and protein biogenesis is interesting in its own right, careful consideration of a potential host's membrane lipid composition may also allow optimization of the yield and activity of membrane proteins that are expressed in a heterologous organism. Here, we review and discuss some examples that illustrate the interdependence between bilayer lipids and the biogenesis of integral membrane proteins. PMID- 17111138 TI - Optimization of the solvent-tolerant Pseudomonas putida S12 as host for the production of p-coumarate from glucose. AB - A Pseudomonas putida S12 strain was constructed that is able to convert glucose to p-coumarate via the central metabolite L: -tyrosine. Efficient production was hampered by product degradation, limited cellular L: -tyrosine availability, and formation of the by-product cinnamate via L: -phenylalanine. The production host was optimized by inactivation of fcs, the gene encoding the first enzyme in the p coumarate degradation pathway in P. putida, followed by construction of a phenylalanine-auxotrophic mutant. These steps resulted in a P. putida S12 strain that showed dramatically enhanced production characteristics with controlled L: phenylalanine feeding. During fed-batch cultivation, 10 mM (1.7 g l(-1)) of p coumarate was produced from glucose with a yield of 3.8 Cmol% and a molar ratio of p-coumarate to cinnamate of 85:1. PMID- 17111139 TI - Online recovery of nisin during fermentation and its effect on nisin production in biofilm reactor. AB - An online removal of nisin by silicic acid coupled with a micro-filter module was proposed as an alternative to reduce detrimental effects caused by adsorption of nisin onto producer, enzymatic degradation by protease, and product inhibition during fermentation. In this study, silicic acid was successfully used to recover nisin from the fermentation broth of Lactococcus lactis subsp. lactis NIZO 22186. The effect of pH (at 6.8 and 3.0) during adsorption process and several eluents (deionized water, 20% ethanol, 1 M NaCl, and 1 M NaCl + 20% ethanol) for desorption were evaluated in a small batch scale. Higher nisin adsorption onto silicic acid was achieved when the adsorption was carried out at pH 6.8 (67% adsorption) than at pH 3.0 (54% adsorption). The maximum recovery was achieved (47% of nisin was harvested) when the adsorption was carried out at pH 6.8 and 1 M NaCl + 20% ethanol was used as an eluent for desorption. Most importantly, nisin production was significantly enhanced (7,445 IU/ml) when compared with the batch fermentation without the online recovery (1,897 IU/ml). This may possibly be attributed to preventing the loss of nisin due the detrimental effects and a higher biomass density achieved during online recovery process, which stimulated production of nisin during fermentation. PMID- 17111140 TI - Biotransformation of gallic acid by Beauveria sulfurescens ATCC 7159. AB - Preparative-scale fermentation of gallic acid (3,4,5-trihydroxybenzoic acid) (1) with Beauveria sulfurescens ATCC 7159 gave two new glucosidated compounds, 4-(3,4 dihydroxy-6-hydroxymethyl-5-methoxy-tetrahydro-pyran-2-yloxy)-3-hydroxy-5-methoxy benzoic acid (4), 3-hydroxy-4,5-dimethoxy-benzoic acid 3,4-dihydroxy-6 hydroxymethyl-5-methoxy-tetrahydro-pyran-2-yl ester (7), along with four known compounds, 3-O-methylgallic acid (2), 4-O-methylgallic acid (3), 3,4-O dimethylgallic acid (5), and 3,5-O-dimethylgallic acid (6). The new metabolite genistein 7-O-beta-D-4''-O-methyl-glucopyranoside (8) was also obtained as a byproduct due to the use of soybean meal in the fermentation medium. The structural elucidation of the metabolites was based primarily on 1D-, 2D-NMR, and HRFABMS analyses. Among these compounds, 2, 3, and 5 are metabolites of gallic acid in mammals. This result demonstrated that microbial culture parallels mammalian metabolism; therefore, B. sulfurescens might be a useful tool for generating mammalian metabolites of related analogs of gallic acid (1) for complete structural identification and for further use in investigating pharmacological and toxicological properties in this series of compounds. In addition, a GRE (glucocorticoid response element)-mediated luciferase reporter gene assay was used to initially screen for the biological activity of the 6 compounds, 2-6 and 8, along with 1 and its chemical O-methylated derivatives 9 13. Among the 12 compounds tested, 11-13 were found to be significant, but less active than the reference compounds of methylprednisolone and dexamethasone. PMID- 17111141 TI - Novel bacterial sulfur oxygenase reductases from bioreactors treating gold bearing concentrates. AB - The microbial community and sulfur oxygenase reductases of metagenomic DNA from bioreactors treating gold-bearing concentrates were studied by 16S rRNA library, real-time polymerase chain reaction (RT-PCR), conventional cultivation, and molecular cloning. Results indicated that major bacterial species were belonging to the genera Acidithiobacillus, Leptospirillum, Sulfobacillus, and Sphingomonas, accounting for 6.3, 66.7, 18.8, and 8.3%, respectively; the sole archaeal species was Ferroplasma sp. (100%). Quantitative RT-PCR revealed that the 16S rRNA gene copy numbers (per gram of concentrates) of bacteria and archaea were 4.59 x 10(9) and 6.68 x 10(5), respectively. Bacterial strains representing Acidithiobacillus, Leptospirillum, and Sulfobacillus were isolated from the bioreactors. To study sulfur oxidation in the reactors, pairs of new PCR primers were designed for the detection of sulfur oxygenase reductase (SOR) genes. Three sor-like genes, namely, sor (Fx), sor (SA), and sor (SB) were identified from metagenomic DNAs of the bioreactors. The sor (Fx) is an inactivated SOR gene and is identical to the pseudo-SOR gene of Ferroplasma acidarmanus. The sor (SA) and sor (SB) showed no significant identity to any genes in GenBank databases. The sor (SB) was cloned and expressed in Escherichia coli, and SOR activity was determined. Quantitative RT-PCR determination of the gene densities of sor (SA) and sor (SB) were 1,000 times higher than archaeal 16S rRNA gene copy numbers, indicating that these genes were mostly impossible from archaea. Furthermore, with primers specific to the sor (SB) gene, this gene was PCR-amplified from the newly isolated Acidithiobacillus sp. strain SM-1. So far as we know, this is the first time to determine SOR activity originating from bacteria and to document SOR gene in bioleaching reactors and Acidithiobacillus species. PMID- 17111142 TI - Lenalidomide in the context of complex karyotype or interrupted treatment: case reviews of del(5q)MDS patients with unexpected responses. AB - Lenalidomide has particular activity in patients with transfusion-dependent del(5q) myelodysplastic syndromes (MDS), but mechanistic information is limited regarding the relationship between erythroid and cytogenetic responses. We reviewed medical records from three distinct subgroups of del(5q) MDS patients who had unexpected effects with lenalidomide treatment: 1. two patients with complex karyotypes who achieved both cytogenetic remissions and transfusion independence; 2. two patients with 5q- syndrome who took lenalidomide for less than 12 weeks but remained transfusion independent for 15+ months still displaying del(5q) metaphases after 6 and 12 months; and 3. one patient who was a non-responder on lenalidomide during treatment but became transfusion independent for 13+ months after discontinuation. All but the latter patient in this series had reduction of affected metaphases, suggesting that erythroid responses might be mediated by result from partial or complete suppression of the malignant clone, either directly or indirectly through modulation of the bone marrow microenvironment. These clinical observations illustrate the heterogeneity of del(5q)MDS pathogenesis and the diversity of lenalidomide responses within this patient subset. PMID- 17111143 TI - Reemergence of JAK2 V617F clone heralds extramedullary leukemia relapse after BMT for transformed essential thrombocytosis. PMID- 17111144 TI - Lyapunov function and the basin of attraction for a single-joint muscle-skeletal model. AB - This paper provides an explicit Lyapunov function for a general single-joint muscle-skeletal model. Using this Lyapunov function one can determine analytically large subsets of the basin of attraction of an asymptotically stable equilibrium. Besides providing an analytical tool for the analysis of such a system we consider an elbow model and show that the theoretical predictions are in agreement with experimental results. Moreover, we can thus distinguish between regions where the self-stabilizing properties of the muscle-skeletal system guarantee stability and regions where nerval control and reflexes are necessary. PMID- 17111145 TI - Eradicating vector-borne diseases via age-structured culling. AB - We derive appropriate mathematical models to assess the effectiveness of culling as a tool to eradicate vector-borne diseases. The model, focused on the culling strategies determined by the stages during the development of the vector, becomes either a system of autonomous delay differential equations with impulses (in the case where the adult vector is subject to culling) or a system of nonautonomous delay differential equations where the time-varying coefficients are determined by the culling times and rates (in the case where only the immature vector is subject to culling). Sufficient conditions are derived to ensure eradication of the disease, and simulations are provided to compare the effectiveness of larvicides and insecticide sprays for the control of West Nile virus. We show that eradication of vector-borne diseases is possible by culling the vector at either the immature or the mature phase, even though the size of the vector is oscillating and above a certain level. PMID- 17111146 TI - Intravitreally administered bevacizumab (Avastin) in minimally classic and occult choroidal neovascularization secondary to age-related macular degeneration. AB - BACKGROUND: Anti-vascular endothelial growth factor (anti-VEGF) agents have been shown to be effective in the treatment of neovascular age-related macular degeneration (AMD). Efficacy and safety of intravitreally administered bevacizumab (Avastin), a humanized monoclonal anti-VEGF, was assessed in minimally classic and occult subfoveal choroidal neovascularization (CNV) due to AMD. METHODS: A prospective interventional study was carried out. Bevacizumab (1.25 mg) was administered intravitreally on a 6-week basis until macular edema, subretinal fluid, and/or pigment epithelial detachment had resolved. Administration was repeated in case of relapse. Ophthalmic evaluations included a complete ophthalmic examination, measurement of the visual acuity (VA), optical coherence tomography, and fluorescein angiography. Main outcome measures were the changes between baseline and last follow-up visit in best-corrected VA, central foveal thickness (CFT) and total macular volume (TMV). RESULTS: From 102 patients [mean age (range) 74.8 (61-85) years], 102 eyes were included. Median (range) duration of follow-up was 18 (6-24) weeks. Statistically significant changes from baseline were observed in best-corrected VA [increase of 1.29 lines (P=0.001)], CFT [reduction of 56 microm (P=0.01)] and TMV [reduction of 0.80 mm(3) (P<0.0001)]. Positive results were obtained in 65/102 (64%) patients after two to three injections as a mean. In a substantial proportion of patients (38%) followed up for at least 18 weeks, recurrence of leakage requiring additional injections was observed. Treatment was well tolerated; two pigment epithelium rips and ten posterior vitreous detachments were reported. CONCLUSIONS: Short term results suggest that intravitreally administered bevacizumab (Avastin) is effective in minimally classic and occult CNV due to AMD. Significant improvements in VA, CFT and TMV were obtained and maintained during follow-up. In some patients, however, recurrence of leakage requiring additional intravitreal injection occurred. Maintenance of the effect of bevacizumab and its safety after repeated and prolonged administration have to be investigated in well-controlled studies. PMID- 17111147 TI - Anterior chamber paracentesis in patients with acute elevation of intraocular pressure. AB - PURPOSE: The purpose was to evaluate the efficacy and the safety of anterior chamber paracentesis (ACP) in acute elevation of intraocular pressure (IOP). METHODS: We prospectively enrolled 20 patients who presented with acute unilateral elevation of IOP above 50 mmHg. IOP was measured before, 10 min, and 1, 7 and 30 days after ACP. The outcome at 1 year was available in 19 patients. RESULTS: The patients included 14 cases of primary acute angle-closure attacks and six cases of secondary glaucoma. IOP decreased from 53.4 +/- 4.2 mmHg to 24.1 +/- 12.5 mmHg at 10 min, to 18.2 +/- 11.1 mmHg at 24 h, and to 16.4 +/- 10.7 mmHg 7 days after ACP. ACP combined with antiglaucomatous medications provided immediate relief of symptoms in all cases and improvement of corneal oedema in 17 cases. We noted three failures in secondary glaucoma. We performed a laser peripheral iridotomy (LPI) in 14 patients and a surgical procedure in 17 eyes. No complications related to ACP were observed. CONCLUSION: ACP is a safe and effective procedure for acute elevation of IOP in acute primary primary angle closure but only remains an add-on therapy to usual treatments. However, this technique must be evaluated in larger series. PMID- 17111148 TI - Transscleral local resection versus iodine brachytherapy for uveal melanomas that are large because of tumour height. AB - BACKGROUND: To compare visual outcome and ocular complications of transscleral local resection (TSR) with those of iodine-125 plaque brachytherapy (IBT) for uveal melanomas categorised as large because of tumour height. METHODS: Retrospective study of 87 patients with a uveal melanoma or=5 microV) were measured in ten normal subjects under conditions of induced optical blur (0 to +3 dioptres) and the relationship to Snellen VA established. The data from 100 consecutive patients (167 eyes) referred for possible non-organic visual loss (NOVL) and 20 patients with confirmed visual pathway dysfunction were reviewed in relation to the results in normal subjects. RESULTS: Snellen VA, under conditions of blur, could be predicted in normal subjects from the check size and contrast required to elicit a criterion PappVEP. These data were tabulated and a quantitative guideline established for the estimation of VA in the patients referred with suspected NOVL. Most (88%) patients referred with suspected NOVL had normal electrophysiology and PappVEPs consistent with normal Snellen VA. In others, they suggested a degree of non-organic overlay. In 20 cases of organic visual loss, PappVEPs were in close agreement with subjective VA. CONCLUSIONS: The short duration pattern onset visual-evoked potential is confirmed as a clinically useful tool in the objective assessment of patients with suspected non organic visual loss. PMID- 17111153 TI - Utility of MLPA in deletion analysis of GCH1 in dopa-responsive dystonia. AB - We applied multiple ligation-dependent probe amplification (MLPA) to patients from three families with characteristic dopa-responsive dystonia (DRD) but no base change in the gene GCH1. We found a complete deletion of GCH1 in affected members of family 1, and partial deletions in affected individuals of family 2 (exons 4-6) and of family 3 (exons 2-6). The findings were confirmed by quantitative real-time PCR. Our investigations demonstrate the utility of MLPA for routine deletion analysis of GCH1 in DRD patients with no sequence changes in this gene. PMID- 17111154 TI - Morphometric analyses of the cervical superior facets and implications for facet dislocation. AB - The articular facets of the cervical spine have been well addressed; however, little information is available on the relationship of the superior facets of the cervical spine to traumatic dislocation in the literature. Morphometric analyses of the superior facets of 30 dried cervical spines from C3 to C7 were performed to determine any morphological differences. The angle of the superior facet with respect to the transverse plane was also measured on computed tomography (CT) scans of 30 patients having neck injury without fracture/dislocation. The vertical and surface lengths of the superior facet were significantly lower (P < 0.01) at C6-C7 levels than at C3-5 levels. The anteroposterior diameter of the superior facet was smaller (P < 0.05) at C6 and C7 levels compared to C3-5 levels. Although the superior facet joint surface is in a more coronal orientation in lower cervical vertebrae, the inclination of the superior facet is more horizontal relative to the transverse plane when measured in vivo. A combination of lower height, smaller anteroposterior diameter of the superior facet, and a more horizontally oriented superior facet at C6 and C7 levels in vivo may explain the predilection of translation relative to one another in the lower cervical spine. PMID- 17111155 TI - Combining mid-trimester maternal plasma homocysteine with uterine artery doppler velocimetry: is it useful? AB - OBJECTIVE: To investigate the possible association between mid-trimester maternal plasma homocysteine concentration, uterine artery Doppler measurements in a two stage screening strategy, and outcome of pregnancy. MATERIALS AND METHODS: This prospective observational study was conducted on healthy women undergoing screening for pre-eclampsia by uterine artery Doppler velocimetry at 20-22 and 24 26 weeks of gestation. Abnormal uterine artery blood flow was defined as an average resistance index (RI) > 0.58 and/or bilateral early diastolic notch. Homocysteine measurement was performed by two competitive immunoassay methods involving two steps at 20-22 and 24-26 weeks' gestation. RESULTS: Sixty women enrolled. Abnormal Doppler findings were found in 18 of 60 (30%) women at 20-22 weeks of gestation. This proportion was reduced to 10% (6/60) at 24-26 weeks of gestation, and two of these six women developed pre-eclampsia later in pregnancy. There was no significant difference in the maternal plasma homocysteine levels in women with abnormal Doppler findings when compared with controls at first and second visits (p > 0.05). CONCLUSION: Mid-trimester maternal homocysteine concentration is not elevated in women with abnormal uterine artery Doppler findings in a two-stage screening method. PMID- 17111156 TI - Efficacy of Yasmin in premenstrual symptoms. AB - OBJECTIVE: To evaluate the effect of oral contraception formulation with drospirenone (Yasmin) on premenstrual symptoms. MATERIALS AND METHODS: An open label non-comparative clinical trial was conducted. One hundred women who desired oral contraception for at least 6 months were recruited. The subjects received a blister pack which contained 21 tablets of 3 mg drospirenone/30 microg ethinyl estradiol for the first four cycles (1 cycle = 28 days). Cycle-5 and -6 blister packs were dispensed during the next visit in cycle 4. The subjects were evaluated on menstruation-related symptoms by using the women's health assessment questionnaire (WHAQ) at baseline and at each follow-up visit for three phases of their menstrual cycle. The measured outcome was the mean score changes from baseline to cycle 6 of WHAQ categories. RESULTS: Of the total 100 subjects, 92 (92%) completed the study. At the premenstrual phase, a significant decrease was seen from baseline to cycle 6 in the mean WHAQ scores for impaired concentration, water retention, negative effect, increased appetite, feeling of well-being and undesirable hair change. At the menstrual phase, a significant decrease was seen from baseline to cycle 6 regarding the mean WHAQ scores for impaired concentration, negative effect, feeling of well-being, and undesirable hair change. At the postmenstrual phase, significant changes were consistently observed regarding the feeling of well-being and undesirable hair change. CONCLUSIONS: Oral contraception formulation with drospirenone (Yasmin) has an ameliorating effect on premenstrual symptoms. This oral contraception may have a role in women who are looking forward to use contraception and also suffer from premenstrual symptoms. PMID- 17111157 TI - [The diagnostic dilemma of acute thoracic pain]. AB - Acute aortic dissection is gaining recognition in Western societies, and is being diagnosed with increasing frequency. New diagnostic imaging modalities, longer life expectancy in general, as well as the increase in the number of hypertension patients have all contributed to the growing awareness of aortic dissections. Compared with acute coronary syndrome and lung embolism, aortic dissection is among the most frequently diagnosed life-threatening conditions involving chest pain. Here we report the case of a 59 year old patient suffering from hypertension and discuss the latest diagnostic and therapeutic procedures in the setting of acute chest pain. PMID- 17111158 TI - [Depressive disorders. A diagnostic and therapeutic challenge also for primary care]. AB - Diagnosis and treatment of depressive disorders play a major role in primary care. Due to their high prevalence in the general population with a pronounced psychiatric and somatic morbidity and a significant subsequent psychosocial disability, such disorders constitute a serious disease entity. This review provides basic knowledge on epidemiology, diagnosis, pharmacotherapy and psychotherapy to enable successful primary care of this important group of patients. PMID- 17111159 TI - Growth and PTH in prepubertal children on long-term dialysis. AB - Growth failure is an important complication for children on dialysis. One possible influence on growth is renal bone disease. We reviewed the case notes of 35 children (23 boys), mean (range) age at inclusion 2.8 (0.25-8.9) years (17 children age <2 years), on dialysis for 2.0 (1-4.8) years, for growth, PTH, calcium and phosphate levels and medications. Data collection ended at age 10 years, commencement of growth hormone (rhGH) or renal transplantation. The mean (range) height standard deviation score (HtSDS) at the start of dialysis was 2.06 (-5.90 to 0.63). No change in HtSDS per year was observed; the median was 0.06 (-1.07 to 2.39). Children aged <2 years showed catch-up growth in the first year on dialysis; median change in HtSDS was 0.31 (-0.78 to 3.13). Mean plasma calcium and ionised calcium were approximately at the mid-point and phosphate just above the mid-point of the respective normal ranges. The median PTH level was 1.52 times the upper limit of normal and levels did not correlate with growth. Our results indicate that intensive nutritional therapy and phosphorus control aiming to keep PTH within the normal range prevents further loss of HtSDS in short children on dialysis. In some children under 2 years of age catch-up growth can be observed in the first dialysis year. PMID- 17111160 TI - Renal caveolin-1 expression in children with unilateral ureteropelvic junction obstruction. AB - Caveolae are plasma membrane invaginations that contain a variety of signal transduction molecules and receptors for growth factors and cytokines. This study was performed to examine the in vivo expression and localization of caveolin-1 in kidneys from 19 children who underwent surgery release of ureteropelvic junction obstruction (UPJO) in relation to renal function and degree of tubulointerstitial fibrosis. Renal biopsies were carried out at the time of surgery for obstruction release. Kidney tissue from children of similar age removed because of carcinoma was used as control. Expression of caveolin-1 at the protein level in renal tissue and urine was demonstrated in patients with technetium 99 m labeled diethylene triamine pentaacetate ((99)Tc DTPA) renal scan 28.8+/-2% and increased tubular interstitial fibrosis in seven patients at the time of obstruction release. Colocalization staining of AT(1) angiotensin II receptor with caveolin-1 in basolateral membrane of epithelial tubule cells, enhanced AT(1) messenger ribonucleic acid (mRNA) and decreased endothelial nitric oxide synthase (eNOS), were shown in these patients. In contrast, absence of association of caveolin-1 with AT(1) receptor expression in proximal and collecting tubule membranes with AT(1) receptor mRNA and eNOS mRNA expression near control were demonstrated in 12 patients, with (99)Tc DTPA renal scan 39.7+2.1% and no evidence of tubulointerstitial fibrosis. From our results, the role of caveolin-1 as a factor contributing to the severity of the tubulointerstitial process resulting from obstructive nephropathy could be suggested. PMID- 17111161 TI - Fungal peritonitis in children on peritoneal dialysis. AB - Fungal peritonitis is a rare but serious complication in children on peritoneal dialysis (PD). In this study, risk factors were evaluated, and therapeutic measures were reviewed. A retrospective, multi-centre study was performed in 159 Dutch paediatric PD patients, between 1980 and 2005 (3,573 months). All peritonitis episodes were reviewed. Fungal peritonitis episodes were evaluated based on possible risk factors and treatment strategy. A total of 321 episodes of peritonitis occurred, with 9 cases of fungal peritonitis (2.9%). Candida peritonitis occurred most frequently (78%). Seven patients (78%) had used antibiotics in the prior month. Fungal peritonitis patients had a higher previous bacterial peritonitis rate compared to the total study population (0.13 versus 0.09 episodes/patient*month), with twice as many gram negative organisms. In all fungal peritonitis patients, the PD catheter was removed. In four patients restart on PD was possible. Fungal peritonitis is a rare complication of PD in children, but is associated with high technique failure. The most important risk factors are a high bacterial peritonitis rate, prior use of antibiotics, and previous bacterial peritonitis with gram negative organisms. The PD catheter should be removed early, but in children, peritoneal lavage with fluconazole before removal may be useful to prevent technique failure. PMID- 17111162 TI - New angiographic measurement tool for analysis of small cerebral vessels: application to a subarachnoid haemorrhage model in the rat. AB - INTRODUCTION: Exact quantification of vasospasm by angiography is known to be difficult especially in small vessels. The purpose of the study was to develop a new method for computerized analysis of small arteries and to demonstrate feasibility on cerebral angiographies of rats acquired on a clinical angiography unit. METHODS: A new software tool analysing grey values and subtracting background noise was validated on a vessel model. It was tested in practice in animals with subarachnoid haemorrhage (SAH). A total of 28 rats were divided into four groups: SAH untreated, SAH treated with local calcium antagonist, SAH treated with placebo, and sham-operated. The diameters of segments of the internal carotid, caudal cerebral, middle cerebral, rostral cerebral and the stapedial arteries were measured and compared to direct measurements of the diameters on magnified images. RESULTS: There was a direct correlation between the cross-sectional area of vessels measured in a phantom and the measurements acquired using the new image analysis method. The spread of repeated measurements with the new software was small compared to the spread of direct measurements of vessel diameters on magnified images. Application of the measurement tool to experimental SAH in rats showed a statistically significant reduction of vasospasm in the SAH groups treated with nimodipine-releasing pellets in comparison to all the other groups combined. CONCLUSION: The presented computerized method for analysis of small intracranial vessels is a new method allowing precise relative measurements. Nimodipine-releasing subarachnoidal pellets reduce vasospasm, but further testing with larger numbers is necessary. The tool can be applied to human angiography without modification and offers the promise of substantial progress in the diagnosis of vasospasm after SAH. PMID- 17111164 TI - Host-related variability in arbuscular mycorrhizal fungal structures in roots of Hedera rhombea, Rubus parvifolius, and Rosa multiflora under controlled conditions. AB - The arbuscular mycorrhizal (AM) morphology of three host plant species inoculated with single and mixed fungal culture and the distribution of AM fungal species in roots of the hosts treated with a mixed culture of AM fungi were determined. The aim was to investigate the effect of host plants and AM fungi on AM morphology of coexisting plant species. Noncolonized rooted cuttings of Hedera rhombea (Miq) Bean, Rubus parvifolius L., and Rosa multiflora Thunb. were inoculated with five fungal species as single and mixed culture inocula. The fungal species used were Gigaspora rosea and Scutellospora erythropa, previously isolated from H. rhombea; Acaulospora longula and Glomus etunicatum from R. parvifolius; and Glomus claroideum from both plant species. A few hyphal and arbusculate coils were seen in the mixed culture-inoculated roots of R. parvifolius; all fungal treatments produced this Paris-type AM in H. rhombea and Arum-type AM in R. parvifolius, and R. multiflora indicates that AM morphology is strongly controlled by the identity of the host plants used in this study. AM fungal rDNA was extracted separately from roots of each replicate plant species inoculated with the mixed fungal culture, amplified, cloned, sequenced, and analyzed to determine the AM fungal species and their respective proportions in roots of each plant species. Glomus etunicatum and G. claroideum of the family Glomaceae generally occurred more frequently in R. parvifolius and R. multiflora, which form Arum-types, whereas S. erythropa, of the family Gigasporaceae, was the most frequently detected species in H. rhombea, which produced Paris-type AM. Although the genotype of the plant species used appears to determine the AM morphologies formed, there was preferential association between the hosts and AM fungal inoculants. PMID- 17111165 TI - [Hundred twentieth anniversary of the Berlin Surgical Society. Significance of military surgery]. AB - November 22, 2006 will mark the one hundred twentieth anniversary of the oldest regional surgical society in Germany, which was founded as the Free Association of Berlin Surgeons in 1886. For years, the chairmen were also chairmen of the German Surgical Society (established 1872). Thus they made important contributions to surgery in Germany as a whole. Professors such as Ernst von Bergmann, August Bier, and Ferdinand Sauerbruch furthered the reputation of the Berlin practitioners and German surgery throughout the world. In the states of Berlin and Brandenburg, development and promotion of surgery in the late eighteenth and nineteenth centuries owed much to the Prussian emperor Friedrich Wilhelm I and the necessities of Prussian battlefields (military surgical training). These battlefields also caused the sharp decline in worldwide importance of Berlin surgeons at the end of World War II. The special geopolitical situation of Berlin in post-war Germany constituted a negative turning point in this region, not only for surgery. As a result of the destruction of Berlin, most records and documents of the Berlin Surgical Society were lost. Research conducted in February 2006 revealed 20 membership lists from the founding years (1893-1914) which were presumed to be lost. These lists can now help us restore part of the Society's identity and roots. New insights have been made regarding the composition of the Society. For example, the large number of military surgeons in these lists reflects the spirit of the times around 1900 and emphasizes the importance of military medicine in imperial Germany. PMID- 17111166 TI - [Implantation of matrix-free cartilage transplants in standardized defects in sheep knee joints]. AB - The goal of the current investigation was to make a comparative analysis of regenerative tissue after autologous de novo cartilage transplantation on the femoral condyles of sheep after a chondral defect. One chondral defect measuring 4 mm in diameter was placed in the center of one medial femoral condyle of each of 48 Suffolk sheep. Twelve defects were left to heal spontaneously, 16 defects were covered with periosteal flaps, and 20 defects were filled with autologous de novo cartilage graft. Macroscopic and microscopic assessments were performed at 26 and at 52 weeks. Regeneration was significantly better (p<0.05) in the transplant group than in the control groups at both 26 weeks and 52 weeks. The differences were most evident in the grade of defect filling, cartilage stability, cell distribution, and matrix assessments. Transplantation of immature, autologous de novo cartilage leads to qualitatively better regeneration both macro- and microscopically than does periosteal flap placement alone. The transplanted, immature cartilage tissue undergoes maturation in vivo. The regenerated tissue has hyaline-like features. PMID- 17111167 TI - [The value of the modified Gleason grading system of prostate adenocarcinoma in routine urological diagnostics]. AB - In several consensus conferences of the International Society of Urological Pathology (ISUP), the Gleason grading system of prostatic carcinomas was modified and adapted to the routine histological diagnostics of specimens of core needle biopsies and radical prostatectomies. The main results are the documentation of all histological patterns (primary, secondary, tertiary) and a shifting of the maximal Gleason score of biopsies from 6 to 7a (3+4) and of radical prostatectomies from 6 and 7 to 7a and 7b (4+3). Score 2 to 4 carcinomas do not exist in the peripheral prostate. pT2 prostatic carcinomas with good prognosis have a maximal score of 7a; pT3 carcinomas with poor prognosis have a most frequent score of 7b. The agreement of the Gleason scores of core needle biopsies and radical prostatectomy specimens is more than 80%. Inter- and intraobserver reproducibility is better than after the conventional Gleason grading. The prognostic value of scores 6 and 7a may be similar. The border between low- and high-grade prostatic carcinoma may be probably Gleason score 7a and 7b. The prognostic value of score 6 should be changed to score 7a in the different therapeutic options for prostatic carcinomas. PMID- 17111168 TI - [Pain acceptance - concept and validation of a German version of the chronic pain acceptance questionnaire]. AB - BACKGROUND: Up to now, only an English version of the Chronic Pain Acceptance Questionnaire (CPAQ) has been available for measuring the acceptance of chronic pain. This paper presents and analyzes a German adaptation of this instrument. METHODS: The German scale was tested on 150 patients at the DRK Pain Center in Mainz. Validity was assessed by means of various indicators of pain and psychosocial impairment. In a subgroup (n=50) the association with the heat pain threshold was determined. RESULTS: The factor structure of the German CPAQ scale is largely concordant with the theoretical model. The internal consistency of the total and subscales is 0.84-0.87 (Cronbach's alpha). The factors are closely related to indicators of psychosocial functioning. Associations with the affective dimension of pain are moderate and with the sensory dimension low. No association is found with heat pain thresholds (thermal sensory analyzer). CONCLUSIONS: The German CPAQ scale is a useful German-language instrument for the measurement of acceptance and shows good psychometric properties. The study confirms that acceptance is not an expression of a physiologically based indolence. PMID- 17111169 TI - Caput medusae. PMID- 17111170 TI - [Current aspects in the osteosynthesis of the midface]. PMID- 17111171 TI - Subcellular localization of fumarase in mammalian cells and tissues. AB - Fumarase, a mitochondrial matrix protein, is previously indicated to be present in substantial amounts in the cytosol as well. However, recent studies show that newly synthesized human fumarase is efficiently imported into mitochondria with no detectable amount in the cytosol. To clarify its subcellular localization, the subcellular distribution of fumarase in mammalian cells/tissues was examined by a number of different methods. Cell fractionation using either a mitochondria fraction kit or extraction with low concentrations of digitonin, detected no fumarase in a 100,000 g supernatant fraction. Immunofluorescence labeling with an affinity-purified antibody to fumarase and an antibody to the mitochondrial Hsp60 protein showed identical labeling pattern with labeling seen mainly in mitochondria. Detailed studies were performed using high-resolution immunogold electron microscopy to determine the subcellular localization of fumarase in rat tissues, embedded in LR White resin. In thin sections from kidney, liver, heart, adrenal gland and anterior pituitary, strong and specific labeling due to fumarase antibody was only detected in mitochondria. However, in the pancreatic acinar cells, in addition to mitochondria, highly significant labeling was also observed in the zymogen granules and endoplasmic reticulum. The observed labeling in all cases was completely abolished upon omission of the primary antibody indicating that it was specific. In a western blot of purified zymogen granules, a fumarase-antibody cross-reactive protein of the same molecular mass as seen in the mitochondria was present. These results provide evidence that fumarase in mammalian cells/tissues is mainly localized in mitochondria and significant amounts of this protein are not present in the cytosol. However, these studies also reveal that in certain tissues, in addition to mitochondria, this protein is also present at specific extramitochondrial sites. Although the cellular function of fumarase at these extramitochondrial locations is not known, the appearance/localization of fumarase outside mitochondria may help explain how mutations in this mitochondrial protein can give rise to a number of different types of cancers. PMID- 17111172 TI - Occupancy of dopamine D(1), D (2) and serotonin (2A) receptors in schizophrenic patients treated with flupentixol in comparison with risperidone and haloperidol. AB - RATIONALE: Flupentixol (FLX) has been used as a neuroleptic for nearly 4 decades. In vitro data show comparable affinity to dopamine D(2), D(1) and 5-HT(2A) receptors and recently, FLX showed to be not inferior to risperidone in schizophrenic patients with predominant negative symptomatology, which was implicated with flupentixol's interaction with 5-HT(2A) and/or D(1) receptors. OBJECTIVES: To assess in vivo receptor occupancy (RO) in patients clinically treated with FLX (n = 13, 5.7 +/- 1.4 mg/day) in comparison with risperidone (RIS, n = 11, 3.6 +/- 1.3 mg/day) and haloperidol (HAL, n = 11, 8.5 +/- 5.5 mg/day). MATERIALS AND METHODS: Each patient underwent two PET scans with 3-N [(11)C]methylspiperone (target: frontal 5-HT(2A)), [(11)C]SCH23390 (striatal D(1)) or [(11)C]raclopride (striatal D(2)). RO was calculated as the percentage reduction of specific binding in comparison with healthy controls. RESULTS: D(2) RO under FLX was between 50% and 70%, indicating an ED(50) of about 0.7 ng/ml serum. 5-HT(2A) and D(1)-RO was 20 +/- 10% and 20 +/- 5% (mean, SEM). Under HAL, D(1)-RO was 14 +/- 6% and under RIS not significantly different from zero. CONCLUSIONS: We were able to demonstrate a moderate 5-HT(2A) and D(1) occupancy under clinically relevant doses of flupentixol, albeit lower than expected from in vitro data and clearly below saturation. Therefore, if flupentixol's efficacy on negative symptoms is based on its interaction with 5-HT(2A) and/or D(1) receptors, it should be highly dependent on serum concentration and thus on dosage and metabolism. However, these data suggest that mechanisms other than D(1) or 5-HT(2A) antagonism may contribute to flupentixol's efficacy on negative symptoms. PMID- 17111173 TI - Cardiovascular reactivity to mental stress is not affected by alpha2 adrenoreceptor activation or inhibition. AB - RATIONALE: It has been postulated that cardiovascular reactivity to mental stress varies with tonic central sympathetic nervous system activity, but pharmacological evidence is missing. OBJECTIVE: To test whether modulation of central sympathetic nervous system activity by alpha2-adrenergic agonism and antagonism affects cardiovascular reactivity to mental stress. MATERIALS AND METHODS: On three five-stepped dose/concentration-response study days, 12 healthy male volunteers received intravenous infusions of dexmedetomidine (alpha2 agonist, target plasma concentrations: 0.04-0.32 ng/ml), yohimbine (alpha2 antagonist, doses: 0.016-0.125 mg/kg), and placebo, respectively. During each dose step, subjects performed a 5-Choice Reaction Time Task (CRTT) and a Paced Auditory Serial Addition Task (PASAT) to induce moderate mental stress. Prestress baseline, as well as stress-induced responses of heart rate, and noninvasive finger arterial blood pressure (Finapres) were assessed. RESULTS: Prestress baseline heart rate and blood pressure decreased with increasing doses of dexmedetomidine and increased with increasing doses of yohimbine. However, dexmedetomidine and yohimbine did not affect stress-induced heart-rate and blood pressure changes. CONCLUSIONS: Cardiovascular reactivity to mental stress is not related to pharmacologically manipulated tonic central sympathetic nervous system activity by alpha2-adrenergic agonists and antagonists. These results do not support the assumption that cardiovascular reactivity is an index of tonic central sympathetic nervous system activity. PMID- 17111174 TI - Effects of the FAAH inhibitor, URB597, and anandamide on lithium-induced taste reactivity responses: a measure of nausea in the rat. AB - RATIONALE: The endogenous cannabinoid system plays a vital role in the control of nausea and emesis. Because of the rapid breakdown and hydrolysis of endocannabinoids, such as anandamide, the therapeutic effects may be enhanced by prolonging their duration of action. OBJECTIVE: The present experiment evaluated the potential of various doses of URB597, a fatty acid amide hydrolase (FAAH) inhibitor, alone and in combination with systemic administration of anandamide to modulate the establishment of lithium-induced conditioned taste reactivity responses in rats. MATERIALS AND METHODS: In experiment 1, on the conditioning day, rats first received an injection of 0.3 mg/kg URB597, 0.15 mg/kg URB597, or vehicle and then received a second injection of anandamide (5 mg/kg) or vehicle, before a 3-min exposure of 0.1% saccharin by intraoral infusion. Immediately after the saccharin exposure, the rats were injected with lithium chloride. On each of three test days, rats received a 3-min intraoral infusion of saccharin solution, and the taste reactivity responses were videotaped and monitored. In experiment 2, the effects of pretreatment with the CB(1) antagonist, AM-251, on URB597 and anandamide-induced suppressed aversion was evaluated. RESULTS: Administration of URB597 alone and in combination with anandamide reduced active rejection reactions elicited by a LiCl-paired saccharin solution; both effects were reversed by pretreatment with AM-251, suggesting that they were CB(1) receptor mediated. CONCLUSIONS: The results suggest that prolonging the action of anandamide by pretreatment with the FAAH inhibitor, URB597, suppresses lithium induced nausea in the rat. PMID- 17111175 TI - Effects of protoscoleces and AgB from Echinococcus granulosus on human neutrophils: possible implications on the parasite's immune evasion mechanisms. AB - The factors affecting the innate susceptibility to Echinococcus granulosus infections are largely unknown. We assessed the interaction of healthy human neutrophils with protoscoleces (PSC) and antigen B (AgB) of E. granulosus by analysis of CD11b upregulation and H(2)O(2) production by flow cytometry. PSC induced neutrophil activation, but their viability was not affected. In contrast, no effects were observed with AgB in both assays. Neutrophil-enriched fractions were also incubated with PSC or AgB, and interleukin 8 (IL-8) production was measured by ELISA. Significant increment in IL-8 production was detected only in supernatants from neutrophil-enriched fractions cultured with PSC. The possible effect of a prior incubation with AgB on the phorbol myristate acetate-induced activation was also evaluated. No changes were observed in CD11b expression, but the H(2)O(2) production was significantly reduced in platelet-activating factor (PAF)-primed neutrophils. These results suggest a possible AgB-mediated mechanism of evasion of the host immune response, which would operate upon events of spillage of the fertile hydatid cyst content. PMID- 17111176 TI - A cathepsin L-like cysteine proteinase gene from the protozoan parasite, Cryptobia salmositica. AB - The present study describes the identification of a cathepsin L-like cysteine proteinase gene (CYS) from the hemoflagellate Cryptobia salmositica. Genomic DNA sequence of cysteine proteinase was obtained by genome walking using degenerate primers. Specific primers were designed to amplify the cDNA of cysteine proteinase from mRNA by rapid amplification of cDNA ends-PCR. The open reading frame of CYS is 1,329 bp, with 443 deduced amino acids. Based on the sequence analysis, cysteine proteinase of C. salmositica is similar to the cathepsin L like cysteine proteinase of kinetoplastid parasites such as Leishmania spp. and Trypanosoma spp. The identification of CYS proteinase gene could help to design cysteine proteinase specific inhibitors. Further studies are required to characterize the complete genomic organization of the cysteine proteinase. PMID- 17111177 TI - First report of Cryptosporidium parvum 'ferret' genotype in American mink (Mustela vison Shreber 1777). AB - A total of 51 faecal samples from wild and farmed mink were analysed by a direct immunofluorescence antibody test. Cryptosporidium oocysts were identified in eight, apparently healthy, farmed American mink (Mustela vison). The isolates were identified as Cryptosporidium parvum 'ferret' genotype by PCR-RFLP and sequencing analysis of a 341-base-pair fragment of the Cryptosporidium oocyst wall protein (COWP) gene. This is the first report of Cryptosporidium in American mink. PMID- 17111178 TI - Oral infection of immunocompetent and immunodeficient mice with Balamuthia mandrillaris amebae. AB - Balamuthia mandrillaris is an opportunistic agent of lethal granulomatous amebic encephalitis (GAE). In mice, we have shown that intranasally instilled B. mandrillaris amebae infect the brain via the olfactory nerve pathway. In this study, we raised the question whether this ameba might also reach the brain after an oral/gastrointestinal infection. Immunocompetent (WT) and immunodeficient (RAG) mice received B. mandrillaris amebae by gavage into the esophagus. Mice of both groups became ill and some died (WT 20%, RAG 40%) within 42 days. All orally infected mice revealed B. mandrillaris amebae in the central nervous system. Outwardly intact amebae and/or specific antigen were found widely distributed in various organs and the stool. The data indicate that oral infection with B. mandrillaris leading to GAE is possible. Exit from the gastrointestinal tract and dissemination remains unresolved. Though stool cultures were negative, transmission of this highly pathogenic ameba via stool cannot be ruled out. PMID- 17111179 TI - Hemozoin biocrystallization in Plasmodium falciparum and the antimalarial activity of crystallization inhibitors. PMID- 17111181 TI - Atlantic sturgeons (Acipenser sturio, Acipenser oxyrinchus): American females successful in Europe. AB - Recent molecular data on the maternally inherited mitochondrial (mt) DNA have challenged the traditional view that the now extinct Baltic sturgeon population belonged to the European sturgeon Acipenser sturio. Instead, there is evidence that American sea sturgeon Acipenser oxyrinchus historically immigrated into the Baltic Sea. In this study, we test the hypothesis that A. oxyrinchus introgressed into, rather than replaced, the A. sturio population in the Baltic. We established four single nucleotide polymorphisms (SNPs) in the nuclear MHC II antigen gene with a species-specific SNP pattern. Using an ancient DNA approach and two independent lines of molecular evidence (sequencing of allele-specific clones, SNaPshot), we detected both A. sturio and A. oxyrinchus alleles in the available museum material of the now extinct Baltic sturgeon population. The hybrid nature of the Baltic population was further confirmed by very high levels of heterozygosity. It had been previously postulated that the immigration of the cold-adapted A. oxyrinchus into the Baltic occurred during the Medieval Little Ice Age, when temperature likely dropped below the degree inducing spawning in A. sturio. Under this scenario, our new findings suggest that the genetic mosaic pattern in the Baltic sturgeon population (oxyrinchus mtDNA, sturio and oxyrinchus MHC alleles) is possibly caused by sex-biased introgression where spawning was largely restricted to immigrating American females, while fertilization was predominantly achieved by abundant local European males. The hybrid nature of the former Baltic sturgeon population should be taken into account in the current reintroduction measures. PMID- 17111182 TI - The parasite connection in ecosystems and macroevolution. AB - In addition to their obvious negative effects ("pathogens"), endoparasites of various kinds play an important role in shaping and maintaining modern animal communities. In the long-term, parasites including pathogens are indispensable entities of any ecosystem. To understand this, it is essential that one changes the viewpoint from the host's interests to that of the parasite. Together with geographic isolation, trophic arms race, symbiosis, and niche partitioning, all parasites (including balance strategists, i.e. seemingly non-pathogenic ones) modulate their hosts' population densities. In addition, heteroxenic parasites control the balance between predator and prey species, particularly if final and intermediate hosts are vertebrates. Thereby, such parasites enhance the bonds in ecosystems and help maintain the status quo. As the links between eukaryotic parasites and their hosts are less flexible than trophic connections, parasite networks probably contributed to the observed stasis and incumbency of ecosystems over geologic time, in spite of continuous Darwinian innovation. Because heteroxenic parasites target taxonomic levels above that of the species (e.g. families), these taxa may have also become units of selection in global catastrophies. Macroevolutionary extrapolations, however, are difficult to verify because endoparasites cannot fossilize. PMID- 17111183 TI - Identification of transcripts associated with cell wall metabolism and development in the stem of sugarcane by Affymetrix GeneChip Sugarcane Genome Array expression profiling. AB - Sugarcane is an important crop in tropical regions of the world, producing a very large biomass and accumulating large amounts of sucrose in the stem. In this study, we present the first report of transcript profiling using the GeneChip Sugarcane Genome Array. We have identified transcripts that are differentially expressed in the sugarcane stem during development by expression profiling using the array and total RNA derived from three disparate stem tissues (meristem, internodes 1-3, 8, and 20) from four replicates of the sugarcane variety Q117 grown in the field. We have identified 119 transcripts that were highly differentially expressed with development and have characterised members of the cellulose synthase (CesA) and cellulose synthase-like (Csl) gene families, which displayed coordinated expression during stem development. In addition, we determined that many other transcripts involved in cell wall metabolism and lignification were also co-expressed with members of the CesA and Csl gene families, offering additional insights into the dynamics of primary and secondary cell wall synthesis in the developing sugarcane stem. PMID- 17111184 TI - [Femtosecond laser in refractive surgery]. PMID- 17111185 TI - [Presbyopia treatment using a femtosecond laser]. AB - Presbyopia is by far the most common refractive error worldwide, with no permanent therapeutic option available. All efforts to restore accommodation by the use of surgery have not led to a generally accepted therapy. However, there is evidence from an animal model that the use of a femtosecond (fs) laser might influence the modulus of elasticity in the lens. Fs-laser impulses can create intralenticular disruption in animal eyes as well as human cadaver lenses and improve elasticity. The concept of treating presbyopia with fs-laser requires a new, complex theory combining the optical and the mechanical aspects of accommodation in the eye. Diagnostic tools for measuring optical change in power and geometrical modification as the eye views from far to near are needed to obtain objective clinical data. A non-invasive treatment of presbyopia to restore accommodation might be possible in the future. PMID- 17111186 TI - Retinal function and histopathology in rabbits treated with Topiramate. AB - PURPOSE: To evaluate retinal function and histopathology in rabbits treated orally with the anti-epileptic drug topiramate. METHODS: Six rabbits were treated with a daily oral dose of topiramate during a period of eight months. Six rabbits receiving water served as controls. Blood samples were analyzed for determination of topiramate serum levels in order to ensure successful drug exposition. Standardized full-field electroretinograms (ERGs) were performed before treatment and then at 2, 3 and 8 months during the treatment period. After terminating treatment the rabbits were sacrificed and the morphology of the sectioned retina was studied. RESULTS: After eight months of treatment the full-field ERG demonstrated normal rod function in treated and control rabbits, but the light adapted 30 Hz flicker b-wave amplitude was significantly reduced in the treated rabbits. This was the case for both the light adapted (Wilcoxon signed ranks test, P = 0.046) and the dark adapted (Wilcoxon signed ranks test, P = 0.028) 30 Hz flicker response from the treated rabbits. Retinal immunohistology revealed a severe accumulation of GABA in amacrine cells and in the inner plexiform layer in 4 of 6 treated rabbits compared to the controls. CONCLUSIONS: Topiramate, orally administrated to rabbits, may cause a significant reduction of the retinal function demonstrated by the reduced b-wave amplitude in the full-field ERG, as well as changes in immunohistology characterized by a severe accumulation of GABA in the inner retina. The retinal dysfunction and the morphological changes indicate that topiramat may damage the retina, similarly to vigabatrin (another anti-epileptic drug). PMID- 17111187 TI - Polymorphisms in methylenetetrahydrofolate reductase gene (MTHFR) and the age of onset of sporadic colorectal adenocarcinoma. AB - BACKGROUND AND AIMS: Evidence is accumulating for a role of folate in the aetiology of colorectal cancer (CRC). The methylenetetrahydrofolate reductase (MTHFR) gene, involved in folate metabolism, is polymorphic in humans. Since it is unknown whether the MTHFR C677T and A1298C polymorphisms alter the risk for CRC, this was the aim of our study. MATERIALS AND METHODS: Genomic DNA from 102 sporadic colorectal adenocarcinoma (SCA) patients and 300 controls was analyzed by polymerase chain reaction followed by restriction digestion for the polymorphisms analyses. RESULTS/FINDINGS: The frequencies of MTHFR C677T and A1298C genotypes were similar in patients and controls. Similar overall risks for disease were seen in individuals with the distinct MTHFR genotypes. However, an excess of the MTHFR 677TT and 677CT genotypes was seen in patients under 50 years, compared with patients at an older age (19.2 vs 13.1% and 61.6 vs 39.5%, respectively; P = 0.04). The differences were more prominent when the frequency of the 677TT plus 677CT genotype was seen in both group of patients (80.8 vs 52.6%, respectively; P = 0.01), and in younger patients compared to controls (80.8 vs 52.3%, P < 0.01). Individuals with the combined genotype had 3.82-fold (95% confidence interval, 1.41-10.42) increased risk of developing SCA under 50 years, compared with those harboring the wild-type genotype. INTERPRETATION/CONCLUSION: These results suggest a role for the MTHFR 677TT plus 677CT genotype in increasing SCA diagnosed at a low age in southeastern Brazil, but additional studies with larger sample sizes should be carried out to clarify this issue. PMID- 17111190 TI - Methotrexate based chemotherapy and deferred radiotherapy for primary central nervous system lymphoma (PCNSL): single institution experience. AB - In the following study, we present our experience in the treatment of PCNSL patients using a multi-step schedule combining chemotherapy and deferred radiotherapy. Patients were treated with two modified M-BACOD cycles and then differently according to radiological response For PR, SD and PD patients, chemotherapy was interrupted and radiotherapy initiated immediately (45 Gy Whole brain RT). With CR patients, chemotherapy was continued with a combination of HMTX, VCZ, PCB and HD Ara-C up to a total of nine cycles. In 36 patients suitable for evaluation (2 patients had undergone tumour resection): 69.4% (25 of 36) had a complete response (CR), 19.4% (7 of 36) had a partial response(PR), 8.3% (3 of 36) had stable disease(SD), and 2.7% (one of 36) had progressive disease (PD). The PR, SD and PD patients were immediately treated by radiotherapy. In this cohort of patients, we observed 6 CR, 4 PR and 2 PD, respectively, following radiotherapy. At first relapse, a total of 16 CR patients were treated by radiotherapy for a total dose of 45 Gy. The OS was 42.1 months for the entire group of patients. In CR patients treated at the moment of recurrence by salvage radiotherapy, the TTP (time lasting from histological diagnosis until recurrence of disease before RT) was 28.3 months, with a 43.4% of disease free patients observed at 2 years. The median disease-free time observed after complete response to radiotherapy was 10.5 months. In 16 patients (34%), further progression of disease was observed following radiotherapy. Two patients developed extra-CNS disease in the breast and testis. When taking into account the patients with radiotherapy delayed at recurrence, the OS was 48 months and the survival rates were 70% and 60% at 2 years and 5 years, respectively. PMID- 17111191 TI - Histopathology and clinical outcome of NF1-associated vs. sporadic malignant peripheral nerve sheath tumors. AB - The differences in the clinical course and histopathology of sporadic and neurofibromatosis type 1 (NF1)-associated malignant peripheral nerve sheath tumors (MPNST) were investigated retrospectively. The collective comprised 38 NF1 patients and 14 sporadic patients. NF1 patients were significantly younger at diagnosis (p<0.001) and had a significantly shorter survival time than sporadic patients (median survival 17 months vs. 42 months, Breslow p<0.05). The time interval to local recurrence and metastatic spread was also significantly shorter in NF1 patients (9.4 months vs. 30.0 months, p<0.01; 9.1 months vs. 33.2 months, p<0.001, respectively). In patients with the original histopathological data available (22 NF1 patients, 14 sporadic cases), NF1-associated MPNST showed a significantly higher cellularity compared to sporadic tumors (p<0.001) whereas sporadic MPNST featured a significantly higher pleomorphism (p<0.01). Most importantly, while histopathological variables correlated with French Federation Nationale des Centres de Lutte Contre le Cancer grading in sporadic MPNST, this was not the case for NF1-associated tumors. The differences between NF1 associated and sporadic MPNST in regard to the clinical course and histopathology may reflect some fundamental differences in biology and pathomechanism of the two tumor groups. Our findings indicate the necessity for a separate grading scheme which takes into account the genetic background in NF1 patients. PMID- 17111192 TI - Lens thiol depletion by peroxynitrite. Protective effect of pyruvate. AB - Pyruvate (PY) is known to be a potent scavenger of H(2)O(2 )by undergoing its peroxidative decarboxylation. While doing so, it also inhibits .OH generation, in addition to its direct .OH scavenging effect. We now hypothesize that PY would also be decarboxylated by cleaving the -O-O- bond in peroxynitrite (ONOO(-)) (PN), with the effect of protecting tissues against NO(x )induced damage. We have verified this by measuring (14)CO(2) formation on incubation of 1-(14)C-PY with 3 morpholinosydnonimine (SIN-1). Its protective effect against PN induced thiol depletion was initially assessed by determining its ability to inhibit oxidation of pure GSH. This was further evaluated by incubating lens homogenate with SIN-1 with or without PY. As conceived, PY did inhibit PN induced loss of protein as well as non-protein -SH. The findings therefore appear potentially useful to protect against nitrite induced damage to the lens and other tissues known to occur with aging and certain diseases such as diabetes. PMID- 17111193 TI - Brd2 is a TBP-associated protein and recruits TBP into E2F-1 transcriptional complex in response to serum stimulation. AB - Brd2 is a novel protein kinase and plays a role in cell cycle-responsive transcription. Recent studies show that Brd2 contributes to E2F-1 regulated cell cycle progression. In this process, Brd2 exhibits scaffold or transcriptional adapter functions and mediates recruitment of both E2F-1 transcription factors and chromatin-remodelling activity to the E2F-1-resposive promoter. In the present study, we show that Brd2 is also a TBP-associated protein and a 26 amino acids peptide in the first bromodomain of Brd2 is essential for Brd2-TBP interaction. We found that serum stimulation of serum starved NIH/3T3 cells efficiently induces the formation of the Brd2-E2F-1-TBP complex in vivo. In this process, Brd2 plays a pivotal role in the recruitment of TBP into a E2F-1 transcriptional complex, as tested in overexpression assay and at the endogenous level. Furthermore, the 26 amino acid peptide that mediates Brd2-TBP interaction is proved to be critical for Brd2-dependent transactivation on E2F-1-responsive promoters, and moreover, Brd2 and E2F-1 may cooperatively participate in various serum-induced transactivation processes in Luciferase-reporter assays. Thus taken together, because Brd2 may recruit a HAT in its transactivational complex and E2F 1 has been found to stimulate transcription by recruiting acetyltransferase and cofactors GCN5, we predict that Brd2 and E2F-1 may act in a cooperative way to introduce an optimal environment for TBP binding to the TATA-element of gene promoters. PMID- 17111194 TI - Enhanced expression of adipocyte-type fatty acid binding protein in murine lymphocytes in response to dexamethasone treatment. AB - Fatty acids have a great influence on the process of lymphocyte apoptosis which is considered as a modulating factor of immune response in both humans and animals. However the mechanism underlying the function of fatty acids in the process of lymphocyte apoptosis is not fully understood. In this study we show that the appearance of adipocyte-type fatty acid binding protein (A-FABP) is induced upon administration of dexamethasone (DEX) in both in vivo and cultured lymphocytes, and its distinct nuclear localization occurs in close relation to the DEX-induced apoptosis process. In immunohistochemistry of mouse spleen, A FABP-immunoreactivity starts to occur 3 h after DEX stimulation, and it massively localizes in the nucleus 8 h after the treatment, while no A-FABP immunoreactivity is discerned in the lymphocytes of normal as well as 24 h post injection spleen. In the murine T-cell leukemia CTLL-2 cells, A-FABP immunoreactivity is also induced in both of the cytoplasm and nucleus when the apoptosis is induced by IL-2 retrieval together with DEX treatment, while in the presence of IL-2 A-FABP-immunoreactivity is confined to the cytoplasm with DEX treatment. On the other hand, A-FABP-immunoreactivity is not detected by IL-2 retrieval alone. The present findings altogether suggest that A-FABP and its ligands, fatty acids, play an important role in the process of apoptosis and the immune modulation induced by DEX. PMID- 17111195 TI - Evaluation of oxidative stress in the thrombolysis of pulmonary embolism. AB - BACKGROUND: To analyse leukocyte function parameters and oxidative stress (OS) in patients with acute pulmonary embolism (PE) treated with thrombolytics. METHODS: Fifteen patients undergoing thrombolysis (TL) with ultra-high dose streptokinase (n = 8), or alteplase (tPA) (n = 7) treatment were studied. Blood samples were collected prior to TL, and then 8 h, 1, 3, 5 and 30 days after treatment. Malondialdehyde (MDA), reduced glutathione (GSH), plasma protein sulfhydryl groups (PSH) levels, superoxide dismutase (SOD) and myeloperoxidase enzyme (MPO) activities were measured in plasma or whole blood for monitoring of the OS markers. Production of reactive oxygen species (ROS) in whole blood was measured by luminol dependent chemiluminescence. Flow cytometry was used to determine CD11a, CD18, and CD97 surface antigen expression on leukocytes. RESULTS: The elevated MDA, ROS and MPO, decreased GSH and PSH levels indicated the presence of OS in patients with PE. MDA significantly (P < 0.05) increased, GSH significantly (P < 0.05) decreased following thrombolysis. ROS production peaked on the 3rd and 5th days. TL was accompanied by significant decrease in granulocyte and monocyte CD11a and CD18 as well as in granulocyte CD97 expression (P < 0.05). CONCLUSION: PE led to OS that was augmented following TL. Decreased adhesion molecule expression of circulating leukocytes in the early phase of TL reflects the pathological leukocyte endothelial cell interactions. PMID- 17111196 TI - Nicotine upregulates the expression of P2Y12 on vascular cells and megakaryoblasts. AB - BACKGROUND: P2Y12 is the major platelet receptor that mediates ADP-induced aggregation. P2Y12 is also expressed by vascular cells. The factors that regulate P2Y12 expression have not been determined. Since nicotine (NIC) has effects on platelet activation and vascular function, and because nicotinic and purinerigic receptors may interact, we determined whether nicotine altered P2Y12 expression. METHODS: Four cell lines (human coronary artery endothelial cells, HCAEC; human umbilical vein endothelial cells, HUVEC; human aortic smooth muscle cells, HASMC; and human megakaryoblastic cells, MEG-01) were cultured in the absence or presence of nicotine. Immunoblotting for P2Y12, P2Y2, and actin was performed. RESULTS: Nicotine, at concentrations of 0.1-1.0 microM, induced P2Y12 (but not P2Y2) expression in all the four cell lines. HASMC exhibited the greatest induction with a sixfold mean increase in P2Y12 expression in response to 0.25 microM nicotine. The induction was inhibited by nicotinic acetylcholine receptor antagonists. Healthy smokers were observed to have higher P2Y12 expression in platelet lysates compared to non-smokers. CONCLUSION: Nicotine induces the expression of P2Y12 in vascular cells and megakaryoblasts, and is mediated by nicotinic acetylcholine receptors. Smokers exhibit higher platelet P2Y12, possibly mediated via nicotine. These results may contribute to a better understanding of the effects of cigarette smoking on platelet activation and the vessel wall. CONDENSED ABSTRACT: The factors that regulate the expression of P2Y12, the platelet ADP receptor, have not been determined. Four cell lines (human coronary artery endothelial cells, HCAEC; human umbilical vein endothelial cells, HUVEC; human aortic smooth muscle cells, HASMC; and human megakaryoblastic cells, MEG-01) were cultured in the absence or presence of nicotine. Nicotine, at concentrations of 0.1-1.0 microM, induced P2Y12 expression in all the four cell lines. HASMC exhibited the greatest induction with a sixfold mean increase in P2Y12 expression in response to 0.25 microM nicotine. The induction was inhibited by nicotinic acetylcholine receptor antagonists. Healthy smokers were observed to have higher P2Y12 expression in platelet lysates compared to non-smokers. These results may contribute to a better understanding of the effects of cigarette smoking on platelet activation and the vessel wall. PMID- 17111197 TI - A comprehensive analysis of 12 thrombophilic mutations and related parameters in patients with inflammatory bowel disease: data from Turkey. AB - BACKGROUND: Possible association of inflammatory bowel disease (IBD) with the most common inherited prothrombotic conditions has been the focus of many investigations. Advance in modern molecular biology is expanding the thrombophilia evaluation steadily. We tried to put forward a comprehensive thrombophilic profile in IBD and to see the probable role of this profile in pathogenesis. METHODS: A total of 60 adults (33 patients and 27 healthy controls) were included. We used the CVD-StripAssay which is based on the reverse hybridization principle to identify a total of 12 thrombophilic gene mutations: Factor V R506Q, Factor V H1299R, prothrombin G20210A, Factor XIII V34L, beta Fibrinogen-455 G-A, PAI-1 4G/5G, platelet GPIIIa L33P, MTHFR C677T, MTHFR A1298C, ACE I/D, Apo B R3500Q and Apo E2/E3/E4, respectively. Besides, we evaluated many related blood parameters such as protein C, protein S, AT-III, IL-6, TNF-alpha, Apo-A1, Apo-B100, homocysteine (tHcy) etc. using commercially available assays. RESULTS: The frequencies of genetic polymorphisms were found to be statistically insignificant among patients and controls, except for three: Beta-Fibrinogen-455G A, MTHFR A1298C and ACE-I/D. Two patients with a history of deep venous thrombosis had more than one polymorphism. Patients with MTHFR C677T and MTHFR A1298C gene mutations had a similar mean tHcy levels with controls. Patients with Apolipoprotein B R3500Q and Apolipoprotein E4 gene mutations had similar mean LDL cholesterol levels. Mean total cholesterol and triglyceride levels were similar in patients and controls of Apo E2, E3, E4 alleles. CONCLUSION: Predominantly, the presence of genetic mutations that predispose to hypercoagulable states does not appear to be in correlation with IBD. There was a statistical difference between the proportions of the mutated allele frequencies of Beta-Fibrinogen-455G A, MTHFR A1298C and ACE-I/D in IBD. PMID- 17111198 TI - Specific types of activated Factor XII increase following thrombolytic therapy with tenecteplase. AB - BACKGROUND: Activated Factor XII (XIIa) is believed to participate in a number of pathophysiological processes including inflammation, thrombosis and fibrinolysis. Increasing XIIa levels following thrombolytic therapy have previously been reported. In contrast to other thrombolytics, tenecteplase (TNK-tpa) does not show paradoxical thrombin activation, indicating a lower procoagulant effect of this fibrin-selective thrombolytic agent. Recent research has demonstrated that in-vivo XIIa exists in a number of different types, and the aim of this study was to investigate plasma variations of different types of XIIa following thrombolytic treatment with TNK-tpa. METHODS: Citrated blood samples were obtained from 34 patients admitted with acute ST-elevation myocardial infarction (STEMI) treated with TNK-tpa. Samples were taken immediately prior to treatment, 30-90 min after and 4 days post-treatment. XIIa measurements were performed using 2 ELISA assays designed to preferentially measure different types of XIIa; XIIaA and XIIaR. Both assays utilised a monoclonal antibody 2/215, which is highly specific for XIIa, as the solid phase capture antibody. The assay for XIIaA used a conjugate based on a polyclonal antibody against the entire XIIa molecule, whilst the assay for XIIaR incorporated a reagent to release otherwise unavailable XIIa and used a conjugate based on a monoclonal antibody against beta XIIa. RESULTS: Changes in plasma XIIaA concentration as a result of therapy were more evident than changes in XIIaR concentration. XIIaA showed a significant increase from 67.1 (49.0-84.4) pM to 97.8 (75.5-133.1) pM [median and 25 and 75% percentiles] in the 30-90 min sample (P < 0.001), returning to pre-intervention levels 61.5 (47.5-81.0) pM by day 4. In contrast, no significant change in XIIaR concentration was observed following thrombolytic therapy with TNK-tpa. CONCLUSION: In patients admitted with STEMI, thrombolytic therapy with TNK-tpa resulted in a significant short-lasting increase in specific types of XIIa (namely XIIaA), whereas other types of XIIa (XIIaR) were largely unaffected by this intervention. PMID- 17111199 TI - Genotypes of the cytochrome p450 isoform, CYP2C9, and the vitamin K epoxide reductase complex subunit 1 conjointly determine stable warfarin dose: a prospective study. AB - BACKGROUND: Warfarin has a narrow therapeutic range and wide inter-individual dosing requirements that may be related to functional variants of genes affecting warfarin metabolism (i.e., CYP2C9) and activity (i.e., vitamin K epoxide reductase complex subunit 1-VKORC1). We hypothesized that variants in these two genes explain a substantial proportion of variability in stable warfarin dose and could be used as a basis for improved dosing algorithms. METHODS: Consecutive consenting outpatients (n = 213) with stable INR (2-3) for >1 month were enrolled. Buccal DNA was extracted using a Qiagen mini-column and CYP2C9*2 and VKORC1 genotyping performed by the Taqman 3' nuclease assay. Sequencing for CYP2C9*3, genotyping was done using Big Dye v3.1 terminator chemistry Dose by genotype was assessed by linear regression. RESULTS: Weekly warfarin dose averaged 30.8 +/- 13.9 mg/week; average INR was 2.42 +/- 0.72. CYP2C9*2/*3 genotype distribution was: CC/AA (wild-type [WT]) = 71.4%, CT/AA = 18.3%, CC/AC = 9.4%, and CT/AC = 1%; VKORC1 genotypes were CC (WT) = 36.6%, CT = 50.7%, and TT = 12.7%. Warfarin doses (mg/week) varied by genotype: for CYP2C9, 33.3 mg/week for WT (CC/AA), 27.2 mg/week for CT/AA (P = 0.04 vs. WT), 23.0 mg/week for CC/AC (P = 0.003), and 6.0 mg/week for CT/AC (P < 0.001), representing dose reductions of 18 31% for single and 82% for double variant carriers; for VKORC1: 38.4 mg/week for WT (CC), 28.6 mg/week for CT (P < 0.001 vs. WT), 20.95 mg/week for TT (P < 0.001). In multiple linear regression, genotype was the dominant predictor of warfarin dose (P = 2.4 x 10(-15)); weak predictors were age, weight, and sex. Genotype-based modeling explained 33% of dose-variance, compared with 12% for clinical variables alone. CONCLUSION: In this large prospective study of warfarin genetic dose-determinants, carriage of a single or double CYP2C9 variant, reduced warfarin dose 18-72%, and of a VKORC1 variant by 65%. Genotype-based modeling explained almost one-half of dose-variance. A quantitative dosing algorithm incorporating genotypes for 2C9 and VKORC1 could substantially improve initial warfarin dose-selection and reduce related complications. PMID- 17111200 TI - Osteoprotegerin is not associated with angiographic coronary calcification. AB - Coronary artery calcification may play a significant role in the pathophysiology of plaque progression and healing. We hypothesized that osteoprotegerin, an inhibitor of osteoclastogenesis, may participate in the calcification of coronary plaques or the response to injury after coronary stenting. A prospective registry was performed in 2004. Blood samples from 100 patients undergoing percutaneous coronary intervention (PCI) were obtained before PCI and 24 h after PCI. The concentrations of osteoprotegerin (OPG), C-reactive protein, interleukin-6, and soluble CD40 ligand (sCD40L) were determined by ELISA. Quantitative coronary angiography was performed to define the presence of culprit lesion calcification (CLC). Comparisons among markers of inflammation and tertiles of OPG were stratified with respect to CLC. Patients with CLC (n = 28) compared with no CLC (n = 71) were older (P < 0.01), had lower creatinine clearance (P < 0.01), lower hemoglobin (P = 0.02), and were less likely to smoke (P = 0.04). Patients without CLC were over twice as likely to present with a marker-positive acute coronary syndrome. CLC was associated with less pre-PCI platelet-mediated inflammation as measured by sCD40L (4.65 vs. 7.15 pg/ml, P = 0.05), but not with lower levels of OPG. Inflammatory cytokines increased significantly after PCI for patients with and without CLC. For patients in the highest tertile of OPG at baseline, there was a reduction in OPG after PCI. Systemic osteoprotegerin levels are not associated with angiographic calcification of culprit plaques. For patients with elevated levels of OPG prior to PCI, there is a significant reduction after PCI consistent with a counterregulatory role for OPG. CONDENSED ABSTRACT: Both calcified and non-calcified culprit plaques exhibited a similar inflammatory response to stent-mediated injury. After PCI, osteoprotegerin decreased while proinflammatory cytokines increased, which may be consistent with a counterregulatory role for osteoprotegerin. PMID- 17111201 TI - Left main coronary thrombosis with essential thrombocythemia. AB - Essential thrombocythemia is a disorder that causes persistent increase in the platelet count. The disease is associated with an elevated risk of thrombosis. A 71-year-old woman was diagnosed with left main coronary thrombosis after an angiogram due to stable angina. One week before the angiogram was taken the patient had also been diagnosed with essential thrombocythemia. After appropriate medical treatment for 5 days the patient underwent an excimer laser treatment, which failed in dissolving the thrombus. Before the patient underwent coronary surgery, thrombopheresis was performed in order to reduce the platelet count. After a successful coronary operation the patient improved completely. PMID- 17111202 TI - Platelet count, mean platelet volume, platelet distribution width, and plateletcrit do not correlate with optical platelet aggregation responses in healthy volunteers. AB - BACKGROUND: Recent advances in technology have made it possible to record various platelet indices. There have been many reports about platelet indices and platelet disorders. The aim of this study is to investigate whether anatomical indices have a correlation with functional aggregation responses using optical method in healthy adults and to evaluate the predictive significance of platelet indices over platelet aggregation responses. METHODS: This study was carried on 31 adults whose ages ranging between 20 and 42. Platelet parameters, including platelet count, mean platelet volume, platelet distribution width and plateletcrit were determined in platelet rich plasma using Abbott Cell-Dyn 4.000. Platelet aggregation was induced by adenosine diphosphate, collagen and epinephrine. Optical aggregation was performed using a turbidometric method. RESULTS: We have observed no correlation between any of platelet indices measured and platelet aggregation responses. CONCLUSIONS: As a result, we found no correlation between platelet aggregation responses obtained with optical method and platelet indices proposed as indicators of certain pathologic conditions, and it does not seem possible to use platelet indices as a direct indicator of platelet activation. In conditions where platelet functions should have been assessed, platelet indices alone are inappropriate and further evaluation is necessary with different methods. PMID- 17111203 TI - Streptokinase--the drug of choice for thrombolytic therapy. AB - Thrombosis, the blockage of blood vessels with clots, can lead to acute myocardial infarction and ischemic stroke, both leading causes of death. Other than surgical interventions to remove or by pass the blockage, or the generation of collateral vessels to provide a new blood supply, the only treatment available is the administration of thrombolytic agents to dissolve the blood clot. This article describes a comprehensive review of streptokinase (SK). We discuss the biochemistry and molecular biology of SK, describing the mechanism of action, structures, confirmational properties, immunogenecity, chemical modification, and cloning and expression. The production and physico-chemical properties of this SK are also discussed. In this review, considering the properties and characteristics of SK that make it the drug of choice for thrombolytic therapy. PMID- 17111204 TI - Deficiency in thrombin-activatable fibrinolysis inhibitor (TAFI) protected mice from ferric chloride-induced vena cava thrombosis. AB - Thrombin-activatable fibrinolysis inhibitor (TAFI) is a plasma carboxypeptidase that renders a fibrin-containing thrombus less sensitive to lysis. Since the role of TAFI in thrombus formation is still controversial in mice, our present study was designed to evaluate mice deficient in TAFI (TAFI(-/-)) on FeCl(3)-induced vena cava and carotid artery thrombosis. Parallel studies were carried out in wild-type mice using a potato carboxypeptidase inhibitor (PCI), a selective inhibitor of activated TAFI (TAFIa). Significant reduction in thrombus formation was observed in TAFI(-/-) mice (n = 8, P < 0.05 compared to wild-type littermates) but not in heterozygous (TAFI(+/-)) mice in 3.5% FeCl(3)-induced vena cava thrombosis. A similar effect was observed following treatment with 5 mg/kg bolus plus 5 mg/kg/h PCI in the same venous thrombosis model in C57BL/6 mice (n = 8, P < 0.01 compared to vehicle). No compositional difference was observed for the venous thrombi in TAFI(-/-) and wild-type littermates with or without PCI treatment using histological assessment. In contrast, neither TAFI deficiency nor treatment with PCI showed antithrombotic efficacy in the 3.5% FeCl(3)-induced carotid artery thrombosis model. In a tail transection bleeding time model, both TAFI deficiency and PCI treatment increased bleeding time up to 4.5 and 3.5 times, respectively, over controls (P < 0.05, n = 8). Similar ex vivo fibrinolytic activities were demonstrated for both TAFI deficiency and PCI treatment as enhanced lysis of thrombin-induced plasma clots and lysis of whole blood clot in a thrombelastograph. These data provide direct evidence for the role of TAFIa in vena cava thrombosis without the addition of exogenous thrombolytic in mice. The strong ex vivo fibrinolytic activity of TAFI deficiency or TAFIa inhibition by PCI provides a biomarker of TAFIa inhibition that tracks in vivo antithrombotic efficacy. PMID- 17111205 TI - Myocardial infarction associated with thrombus formation in non-culprit coronary arteries. AB - Acute coronary syndromes may be associated with a systemic acute pro-thrombotic condition, possibly involving inflammatory mechanisms as well, which are not confined to a single spot in the coronary circulation. Multivessel coronary thrombosis appears to be an exceptionally rare clinical finding. Here we present a case of anterior MI complicated by thrombi in circumflex and right coronary arteries. PMID- 17111206 TI - The U.S. Thrombosis and Hemostasis Centers pilot sites program. AB - Venous thromboembolism (VTE) is a common disorder associated with significant morbidity and mortality. Despite important advances in understanding the etiology of VTE, delivery of care to patients with thrombosis and thrombophilia is frequently incomplete and highly variable. A comprehensive model of health care has been used successfully to treat and prevent complications for people with hemophilia and other chronic disorders. The effectiveness of an integrated healthcare model for patients with all coagulation disorders has yet to be evaluated. The Division of Hereditary Blood Disorders of the Centers for Disease Control and Prevention (CDC) is collaborating with eight Thrombosis and Hemostasis Centers (pilot sites) to provide health-related services and conduct research directed toward the reduction or prevention of complications of thrombosis and thrombophilia. The initial objectives of the collaboration are to (1) determine the efficacy of integrated multidisciplinary care and prevention services for people with hemostatic disorders, (2) assess unmet needs for service delivery and identify outreach strategies to improve access to care, (3) develop effective messages aimed at disease management and prevention, and (4) foster the development of training programs to enhance provider skills for the delivery of patient care. To address these objectives, the investigators and CDC have developed and implemented a web-based patient registry to follow prospectively service allocation and patient outcomes. Funding for the program began in October 2001. All eight funded centers are affiliated with U.S. medical schools. Principal investigators at the centers are hematologists (five adult, two pediatric) or cardiologists. Faculty in obstetrics-gynecology, surgery, and multiple other specialties are integral to the model of care at the centers. Other critical components at the centers are clinical laboratory services, training programs, research networks, and education and outreach programs. From August 2003 to March 2006, over 2,600 patients were enrolled in the registry, accounting for a total of more than 5,000 visits to the centers. Immediate goals of the data collection at the centers are to characterize patients receiving care at centers and document the state of health services provided. Long-term goals are to evaluate prospectively clinical outcomes for patients receiving multidisciplinary care and prevention services at centers. The network of data collection across centers will facilitate future collaborative clinical and epidemiologic investigations and enhance collective expertise in hemostasis and coagulation disorders. PMID- 17111207 TI - Phase I trial and antitumor effects of BZL101 for patients with advanced breast cancer. AB - BACKGROUND: Botanical therapies are often used by breast cancer patients yet few clinical trials have evaluated their safety and efficacy. We studied mechanisms of activity and performed a phase I clinical trial in patients with advanced breast cancer to evaluate BZL101, an aqueous extract from Scutellaria barbata. METHODS: Preclinical studies were conducted in vitro to characterize cell death induced by BZL101. In a phase I trial, eligible patients had histologically confirmed, measurable metastatic breast cancer. Treatment consisted of 350 ml per day of oral BZL101, administered as sole cancer therapy until disease progression, toxicity or personal preference to discontinue. Primary endpoints were safety, toxicity and tumor response. RESULTS: BZL101 extract induced strong growth inhibition and apoptosis of breast cancer cell lines. In the phase I trial, 21 patients received BZL101. Mean age was 54 years (30-77) and mean number of prior treatments for metastatic disease was 3.9 (0-10). There were no grade III or IV adverse events (AEs). The most frequently reported BZL101-related grade I and II AEs included: nausea (38%), diarrhea (24%), headache (19%) flatulence (14%), vomiting (10%), constipation (10%), and fatigue (10%). Sixteen patients were evaluable for response. Four patients had stable disease (SD) for >90 days (25%) and 3/16 had SD for >180 days (19%). Five patients had objective tumor regression, one of which was 1 mm short of a PR based on RECIST criteria. CONCLUSIONS: BZL 101 inhibits breast cancer cell lines by inducing apoptosis. In a phase I clinical trial, BZL101 was safe and had a favorable toxicity profile. BZL101 demonstrated encouraging clinical activity in this heavily pretreated population. PMID- 17111208 TI - Pregnancy loss, stigma, irony, and masculinities: reflections on and future directions for research on religion in the global practice of IVF. PMID- 17111210 TI - Coupling of a 3D finite element model of cardiac ventricular mechanics to lumped systems models of the systemic and pulmonic circulation. AB - In this study we present a novel, robust method to couple finite element (FE) models of cardiac mechanics to systems models of the circulation (CIRC), independent of cardiac phase. For each time step through a cardiac cycle, left and right ventricular pressures were calculated using ventricular compliances from the FE and CIRC models. These pressures served as boundary conditions in the FE and CIRC models. In succeeding steps, pressures were updated to minimize cavity volume error (FE minus CIRC volume) using Newton iterations. Coupling was achieved when a predefined criterion for the volume error was satisfied. Initial conditions for the multi-scale model were obtained by replacing the FE model with a varying elastance model, which takes into account direct ventricular interactions. Applying the coupling, a novel multi-scale model of the canine cardiovascular system was developed. Global hemodynamics and regional mechanics were calculated for multiple beats in two separate simulations with a left ventricular ischemic region and pulmonary artery constriction, respectively. After the interventions, global hemodynamics changed due to direct and indirect ventricular interactions, in agreement with previously published experimental results. The coupling method allows for simulations of multiple cardiac cycles for normal and pathophysiology, encompassing levels from cell to system. PMID- 17111211 TI - In vivo MR imaging of tissue-engineered human mesenchymal stem cells transplanted to mouse: a preliminary study. AB - Current progress integrating stem cell biology and tissue engineering techniques has been invaluable to clinical applications. Prior to the application of cellular transplantation technique to patients, we need to establish techniques that can monitor their tissue biodistribution non-invasively. In this study, we proposed an imaging modality using MRI to not only monitor implanted scaffold in vivo, but also to track transplanted cells and behavior around the implant. For this purpose, human bone marrow-derived mesenchymal stem cells (hMSCs) were labeled with superparamagnetic iron oxide (Feridex) and then labeled hMSCs were cultured in a gelatin sponge used as a scaffold to support cell growth and proliferation. Histological assessment and MTT assay showed that cell labeling with MR contrast agent did not harm cell viability. Also, Feridex-labeled hMSCs showed a significant decrease in T2 signal intensity, even within the gelatin sponge in vitro. After implanting the sponge/cell complex in vivo, we could visualize cellular behavior around the implant over time using a noninvasive MRI modality and this finding was correlated with histological study, which illustrates the potential of a new approach proposed here for in vivo monitoring of implanted cell-based tissue-engineered product. PMID- 17111212 TI - Multi-scale computational model of fuel homeostasis during exercise: effect of hormonal control. AB - A mathematical model of the whole-body metabolism is developed to predict fuel homeostasis during exercise by using hormonal control over cellular metabolic processes. The whole body model is composed of seven tissue compartments: brain, heart, liver, GI (gastrointestinal) tract, skeletal muscle, adipose tissue, and "other tissues". Each tissue compartment is described by dynamic mass balances and major cellular metabolic reactions. The glucagon-insulin controller is incorporated into the whole body model to predict hormonal changes during exercise. Moderate [150 W power output at 60% of peak oxygen consumption (VO(2max))] exercise for 60 min was implemented by increasing ATP utilization rates in heart and skeletal muscle. Arterial epinephrine level was given as an input function, which directly affects heart and skeletal muscle metabolism and indirectly other tissues via glucagon-insulin controller. Model simulations were validated with experimental data from human exercise studies. The exercise induced changes in hormonal signals modulated metabolic flux rates of different tissues in a coordinated way to achieve glucose homeostasis, demonstrating the efficacy of hormonal control over cellular metabolic processes. From experimental measurements of whole body glucose balance and arterial substrate concentrations, this model could predict the dynamic changes of hepatic glycogenolysis and gluconeogenesis, which are not easy to measure experimentally, suggesting the higher contribution of glycogenolysis ( approximately 75%). In addition, it could provide dynamic information on the relative contribution of carbohydrates and lipids for fuel oxidation in skeletal muscle. Model simulations indicate that external fuel supplies from other tissue/organ systems to skeletal muscle become important for prolonged exercise emphasizing the significance of interaction among tissues. In conclusion, this model can be used as a valuable complement to experimental studies due to its ability to predict what is difficult to measure directly, and usefulness to provide information about dynamic behaviors. PMID- 17111213 TI - Managed care and the scale efficiency of US hospitals. AB - Managed care penetration has been partly responsible for slowing down increases in health care costs in recent years. This study uses a 1992-1996 Health Care Utilization Project sample of hospitals to analyze the relationship between managed care penetration in local insurance markets and hospital scale efficiency. After controlling for hospital and market area variables, we find that managed care insurance, particularly the preferred provider type, is associated with increases in hospital scale efficiency in tertiary cases. The results presented here are consistent with the view that managed care can lead to reductions in health cost inflation by controlling the diffusion of technology via improvements in the scale efficiency of hospitals. PMID- 17111214 TI - Sexual behavior and drug use among Asian and Latino adolescents: association with immigrant status. AB - This paper contributes new evidence on the association between immigrant status and health by describing and attempting to explain patterns of co-occurring sex and drug use behaviors among Asian and Latino adolescents in the United States. Nine patterns of sex and drug use behaviors were identified from a cluster analysis of data from 3,924 Asian and Latino youth (grades 7-12) who participated in the National Longitudinal Study of Adolescent Health (Add Health). The relationship between immigrant status and risk cluster membership was evaluated with multinomial logistic regression. Compared to foreign-born youth, U.S. born Asian and Latino adolescents were more likely to engage in sex and drug risk behaviors. Family and residential characteristics associated with immigrant status partly accounted for this finding. The results indicate that among Asian and Latino adolescents, assimilation to U.S. risk behavior norms occurs rapidly and is evident by the second generation. PMID- 17111215 TI - The effects of years lived in the United States on the general health status of California's foreign-born populations. AB - The purpose of this study was to examine the effect of immigrant status and years lived in the United States on the general health status of California's foreign born populations. Two models were constructed using California's aggregate adult population and the adult foreign-born population. A binary logistic regression was conducted using data from the 2003 California Health Interview Survey. California's immigrant populations were found to enjoy a self-assessed health advantage compared to California's US-born population. The results present evidence of a downward trend in self-assessed health amongst California's foreign born population associated with increased years lived in country. The initial health advantage found among California's foreign-born appears to function mainly through socio-economic factors. These results suggest that the deterioration in health associated with increased years lived in the US may be an effect of immigrants' exposure to California's environmental determinants of human health rather that the loss of culture-specific protective factors. PMID- 17111216 TI - Evolution of the colored eco-genetic relationship map (CEGRM) for assessing social functioning in women in hereditary breast-ovarian (HBOC) families. AB - The CEGRM was initially conceived as a simple, concise, visual representation of the social interaction domains of information, tangible services and emotional exchanges (Kenen, R., & Peters, J. (2001). J Genet Counsel, 10, 289-309). A blend of the genetic pedigree, genogram, and ecomap, the CEGRM was developed to facilitate contemporary genetic counseling goals. An exploratory pilot study of 20 subjects showed that it was feasible, comfortable and efficiently accomplished, and that the process was useful both for assessment and as an intervention with study participants (Peters, J. A., Kenen, R., Giusti, R., Loud, J., Weissman, N., & Greene, M. H. (2004). Am J Med Genet Part A, 130A, 258-264). Subsequently, we have extended the CEGRM to 150 women from hereditary breast/ovarian cancer (HBOC) families; three different investigators have successfully administered this tool. The preliminary findings from the exploratory study were confirmed in the larger sample. Engaging in the interactive, insight-promoting CEGRM process provides a novel tool for assessing the social context of genetic testing, and helping high-risk women better understand and integrate genetic information into their personal and family identities, health beliefs, and decisions. PMID- 17111217 TI - Associations between methamphetamine use and HIV among men who have sex with men: a model for guiding public policy. AB - Among men who have sex with men (MSM) in Los Angeles County, methamphetamine use is associated with high rates of HIV prevalence and sexual risk behaviors. In four separate samples of MSM who differed in the range of their intensity of methamphetamine use, from levels of recreational use to chronic use to those for MSM seeking drug abuse treatment, the association between methamphetamine use and HIV infection increased as the intensity of use increased. The lowest HIV prevalence rate (23%) was observed among MSM contacted through street outreach who mentioned recent methamphetamine use, followed by MSM who used at least once a month for six months (42%), followed by MSM seeking intensive outpatient treatment (61%). The highest rate (86%) was observed among MSM seeking residential treatment for methamphetamine dependence. The interleaving nature of these epidemics calls for comprehensive strategies that address methamphetamine use and concomitant sexual behaviors that increase risk of HIV transmission in this group already at high risk. These and other data suggest that MSM who infrequently use methamphetamine may respond to lower intensity/lower cost prevention and early intervention programs while those who use the drug at dependence levels may benefit from high intensity treatment to achieve goals of reduced drug use and HIV-risk sexual behaviors. PMID- 17111218 TI - Mental health during pregnancy: a study comparing Asian, Caucasian and Native Hawaiian women. AB - OBJECTIVE: This study examines perinatal mental health issues, ethnic differences, and comorbidity among pregnant women in Hawaii. METHODS: Eighty-four participants were recruited from women, ages 18-35, seeking prenatal care on Oahu. They were interviewed at their initial prenatal visit about substance use, depression, and anxiety. RESULTS: Sixty-one percent of women screened positive for at least one mental health issue. Thirteen percent of all pregnant women reported drinking during pregnancy with 5% reporting problem drinking, 15% reported smoking cigarettes on a regular basis, 5% of pregnant women had probable depression, and 13% of pregnant women had probable anxiety. Significant ethnic differences were found in cigarette smoking, with the highest rate among Native Hawaiian women (35%). Native Hawaiian women were also more likely to binge drink. CONCLUSION: Given the high rates of potential mental health issues in our sample, our findings highlight the importance of screening and treatment for mental health issues early in pregnancy in Hawaii. PMID- 17111219 TI - Interactive effects of leaf damage, light intensity and support availability on chemical defenses and morphology of a twining vine. AB - In a greenhouse study, we evaluated the effect of the light environment and support availability on the induction of tropane alkaloids (TAs) after leaf damage in the twining vine Convolvulus arvensis. We also tested whether leaf damage modifies the phenotypic responses of the plant to shade and physical support. We found a consistent pattern of induction of TAs after leaf damage in each environmental condition. The induction of TAs was differentially affected by combinations of support and light treatments. In the sun, prostrate and climbing vines exhibited similar induced responses. In the shade, prostrate vines showed greater induced responses. Thus, vines showed the greatest chemical induction when damage occurred in a resource-poor environment (shade), and there was no cue (support) of future increase in resource uptake. Damaged plants showed reduced plasticity to shading in leaf shape and internodes and petiole length in comparison with control, undamaged plants. Herbivory and/or induced responses to herbivory may limit adaptive plant responses to the environment. Therefore, the negative consequences of herbivory on plant fitness might be magnified in a context of changing environments. PMID- 17111220 TI - The auxin-like activity of humic substances is related to membrane interactions in carrot cell cultures. AB - A detailed characterization of two humic fractions was performed: One with low relative molecular mass (LMr<3,500 Da) and one with high relative molecular mass (HMr>3,500 Da). Distinct (1)H NMR spectroscopic patterns were observed for the two fractions. HMr showed an aromatic proton region, an intense and broad region (3.0-5.0 ppm) attributed to sugar-like and polyether components, and an intense doublet at 1.33 ppm (identified as protons of the beta-CH(3) in lactate). In contrast, LMr did not show resonances due to aromatic protons and was characterized by a broad unresolved region, assigned to sugar-like components. The (13)C NMR spectra showed that the LMr humic fraction was richer in carboxylic and aliphatic C groups compared to HMr fraction. These substances were fluorescein-labeled [fluorescein isothiocyanate (FITC)], and their interaction with carrot cells in culture was monitored for 10 d, and compared to FITC-indole 3-acetic acid (IAA) to clarify their mechanisms of biological activity. After different incubation times, fluorescein staining of carrot cells and decrease of fluorescein concentration in the culture medium were evaluated. Fluorescent membrane staining was only present in IAA and the LMr humic fraction treated cell cultures. A consequential decrease of fluorescein concentration in the culture media was also observed. Pretreatment of carrot cells with unconjugated IAA or LMr humic fraction markedly reduced fluorescein staining of both FITC-IAA and FITC-LMr humic fraction. Blocking tests gave indirect evidence of possible binding of the LMr humic fraction to IAA cell membrane receptors. These results indicate that the two humic fractions behave differently. Only LMr humic fraction, like IAA, interacts with cellular membranes in carrot cell cultures. PMID- 17111221 TI - Fate of tannins in Corsican pine litter. AB - Tannins are ubiquitous in higher plants and also in litter and soils where they affect many biogeochemical processes. Despite this well-recognized role, their fate in litter and mineral soils is hardly known, as often only trace amounts, if any, are measured. In this study, we conducted an incubation experiment with Corsican pine litter to which known amounts of tannic acid (TA) or condensed tannins (CTs) from Corsican pine were added. Using Folin-Ciocalteu as a measure for total phenolics and HCl-butanol as an assay specific for CTs, acetone/water extractable phenolics and tannins decreased with time towards very low levels. Application of thermally assisted hydrolysis and methylation to litter before and after acetone/water extraction revealed that TA concentration decreased. By contrast, CTs remained to a great extent in the litter and could not be extracted suggesting that they were tightly bound. PMID- 17111222 TI - The mammary bud as a skin appendage: unique and shared aspects of development. AB - Like other skin appendages, the embryonic mammary gland develops via extensive epithelial-mesenchymal interactions. Early stages in embryonic mammary development strikingly resemble analogous steps in the development of hair follicles and teeth. In each case the first morphological sign of development is a localized thickening in the surface epithelium that subsequently invaginates to form a mammary, hair follicle or tooth bud. Similar sets of intersecting signaling pathways are involved in patterning the mammary, hair follicle and dental epithelium, directing placode formation, and controlling bud invagination. Despite these similarities, subsequent events in the formation of these appendages are diverse. The mammary bud extends to form a sprout that begins to branch upon contact with the mammary fat pad. Hair follicles also extend into the underlying mesenchyme, but instead of branching, hair follicle epithelium folds around a condensation of dermal cells. In contrast, teeth undergo a more complex folding morphogenesis. Here, we review what is known of the molecular and cellular mechanisms controlling early steps in the development of these organs, attempt to unravel both common themes and unique aspects that can begin to explain the diversity of appendage formation, and discuss human genetic diseases that affect appendage morphogenesis. PMID- 17111223 TI - Alveolar and lactogenic differentiation. AB - The mouse mammary gland is a complex tissue that proliferates and differentiates under the control of systemic hormones during puberty, pregnancy and lactation. Once a highly branched milk duct system has been established, during mid/late pregnancy, alveoli, little saccular outpouchings, sprout all over the ductal system and differentiate to become the sites of milk secretion. Here, we review the emerging network of the signaling pathways that connects hormonal stimuli with locally produced signaling molecules and the components of intracellular pathways that regulate alveologenesis and lactation. The powerful tools of mouse genetics have been instrumental in uncovering many of the signaling components involved in controlling alveolar and lactogenic differentiation. PMID- 17111224 TI - Nerve growth factor differentially affects spatial and recognition memory in aged rats. AB - In rats, object discrimination depends on the integrity of the cholinergic system, thus it could be expected that nerve growth factor (NGF) can improve the behavior in aged subjects. The interactive effect of age and cholinergic improvement was assessed behaviorally in young and aged rats. Animals were injected by infusion of NGF into the lateral ventricles and they were tested in two behavioral tasks: an object-location and an object-recognition task. Spatial and recognition memory were assessed in an open field containing five different objects. Rats were submitted to six consecutive sessions. Both age-groups showed comparable habituation of exploratory response in Session 1-4. Discrimination index (DI) was calculated to assess responses to spatial change in Session 5 and object change in Session 6. Control young and aged rats were able to discriminate between familiar and novel object, however DI was lower in aged rats. Treatment with NGF induced decline of object discrimination in both age-groups. Different results were obtained in spatial displacement test. NGF was able to improve spatial memory in aged rats, but had no effect in young controls. These data confer on NGF potential role in improving spatial but not episodic memory in aged rats. PMID- 17111225 TI - Transport of living cells with magnetically assembled nanowires. AB - We present a technique of transporting and positioning living cells internalized by nickel (Ni) nanowires guided by magnetic field. Nanoscale magnetic nanowires are internalized by the Rat Neuroblastoma (ATCC number CRL-2754) and the cells are transported and positioned by magnetic fields from the magnetic material coated electrodes. This technique may enable the interfacing between neurons and electronic devices to empower investigations pertaining to non-invasive neuron probing as well as nanofabricated neural pharmacological technologies. PMID- 17111226 TI - Voltage-sensitive ion channels and cancer. AB - Plasma membrane voltage-sensitive ion channels classically have been associated with a variety of inherited diseases or "channelopathies" that range in the severity of symptoms from mild to lethal. Ion channels are found throughout the body and are responsible for facilitated diffusion of ions down the electrochemical gradient across cells membranes in various tissues. Voltage sensitive ion channels open in response to changes in the membrane potential and are primarily found in excitable cells and tissues. Potassium, calcium, and sodium channels play critical roles in the development of major diseases, such as hyperkalemia, epilepsy, congenital myotonia and several cardiac arrythmias. Recently, cancer studies have begun to define the role of voltage-sensitive ion channels in the progression of cancer to a more malignant phenotype. In cancer, the increased expression or increased kinetics of voltage-sensitive ion channels is associated with an increasing malignant potential as evinced by their role in cell proliferation, migration and survival; as such, these channels are becoming the targets of significant drug development efforts to block or reduce voltage sensitive ion channel activity in order to prevent or combat malignant disease. PMID- 17111227 TI - Microporated PEG spheres for fluorescent analyte detection. AB - Poly(ethylene glycol) (PEG) hydrogels have been used to encapsulate fluorescently labeled molecules in order to detect a variety of analytes. The hydrogels are designed with a mesh size that will retain the sensing elements while allowing for efficient diffusion of small analytes. Some sensing assays, however, require a conformational change or binding of large macromolecules, which may be sterically prohibited in a dense polymer matrix. A process of hydrogel microporation has been developed to create cavities within PEG microspheres to contain the assay components in solution. This arrangement provides improved motility for large sensing elements, while limiting leaching and increasing sensor lifetime. Three hydrogel compositions, 100% PEG, 50% PEG, and microporated 100% PEG, were used to create pH-sensitive microspheres that were tested for response time and stability. In order to assess motility, a second, more complex sensor, namely a FITC-dextran/TRITC-Con A glucose-specific assay was encapsulated within the microspheres. PMID- 17111228 TI - Derivation and left ventricular pressure phase plane based validation of a time dependent isometric crossbridge attachment model. AB - Huxley's crossbridge attachment model predicts tension (contractile force) development in isometric (fixed length) cells using constant attachment and detachment rates. Alternative models incorporating time-varying calcium concentrations are complex (coupled linear differential equations) and use time dependent inputs (calcium, elastance, etc.) to model multiple states. We hypothesize that by incorporating the known significant rise and fall in intracellular calcium, via either an asymmetric damped function or a symmetric Gaussian function, into a time-varying, rather than constant, attachment rate function, the Huxley model prediction for tension (i.e., chamber pressure) in isovolumic (isometric) non-ejecting beats will improve. To test the hypothesis that the time-dependent model-predicted (TDM) pressure fits the in vivo isometric (isovolumic) LV pressure phase-plane (PPP) contour better than the constant attachment rate predicted pressure, we used the TDM to fit non-ejecting, premature ventricular contraction (PVC) PPP contours in 6 subjects. Conventional model fit was poor (relative error 74.0%+/-12.5%), while the asymmetric damped TDM rate function provided slight improvement relative to the conventional time independent model (relative error 55.4%+/-9.8%). The symmetric Gaussian rate function TDM provided the best PPP fit to all non-ejecting beats tested (relative error 19.8%+/-4.8%). We conclude that approximating the lumped attachment rate via a time-varying, rather than constant, rate function generates a physiologically viable model of crossbridge behavior. The PPP provides the optimal arena for alternate mathematical formulation assessment of LVP contour prediction by time-dependent attachment rate functions and facilitates modeling of cardiac contraction and relaxation. PMID- 17111229 TI - Being silenced: the impact of negative social reactions on the disclosure of rape. AB - Rape survivors who speak out about their assault experiences are often punished for doing so when they are subjected to negative reactions from support providers. These negative reactions may thereby serve a silencing function, leading some rape survivors to stop talking about their experiences to anyone at all. The current study sought to examine this worst case scenario. Focusing on the qualitative narratives of eight rape survivors who initially disclosed the assault but then stopped disclosing for a significant period of time, this study sought to provide an in-depth description of how negative reactions silenced these survivors. Three routes to silence were identified: 1) negative reactions from professionals led survivors to question whether future disclosures would be effective; 2) negative reactions from friends and family reinforced feelings of self-blame; and 3) negative reactions from either source reinforced uncertainty about whether their experiences qualified as rape. Implications for future research and practice are discussed. PMID- 17111230 TI - The social tariff of EQ-5D is not adequate to assess quality of life in patients with low back pain. AB - BACKGROUND: We compared the quality of life perceived by patients with non specific low back pain with that predicted by the social tariff of the Spanish version of EQ-5D questionnaire. METHODS: For each health state of the EQ-5D, an adjusted tariff for patients with back pain was obtained using a linear regression model in which the linear effect of the three levels of response for each of the five domains of the EQ-5D was assumed. These coefficients were compared with those obtained for the general Spanish population. In another model, equal in structure to the standard "Dolan N3" model, the linear effect of the five domains was not assumed. RESULTS: In 633 patients, 93 health states were recorded. Significant differences in the coefficients of self-care (p = 0.003) and the maximum level of severity in any dimension (p < 0.0001) were observed. The social tariff of the healthy population is different from the tariff of low back pain patients, with general population values being lower than those of patients, particularly in the 211 health states in which any dimension is at level 3. Weights of the different EQ-5D dimensions showed a non-linear effect on the patients' quality of life. CONCLUSION: Methods used to develop the social tariff for the Spanish version of EQ-5D were inadequate. In addition, this study shows that values given by the general population are different from those of low back pain patients, further confirming that the social tariff of EQ-5D should not be used with actual patients. PMID- 17111231 TI - Reliability and validity of the Infant and Toddler Quality of Life Questionnaire (ITQOL) in a general population and respiratory disease sample. AB - OBJECTIVE: To evaluate feasibility, internal consistency, test-retest reliability, and concurrent and discriminative validity of the Infant and Toddler Quality of Life Questionnaire (ITQOL) for parents of pre-school children with 12 scales (103-items) covering physical and psychosocial domains and impact of child health on parents, in comparison with the TNO-AZL Pre-school Children Quality of Life Questionnaire (TAPQOL). METHODS: Parents of children from a random general population sample (2 months-4 years old; n = 500) and of an outpatient clinic sample of children with respiratory disease (5 months-[Formula: see text] years old; n = 217) were mailed ITQOL and TAPQOL questionnaires; a retest was sent after two weeks. RESULTS: Feasibility: The response was >or=80% with few missing and non-unique ITQOL-answers (<2%) in both study populations. Some ITQOL-scales (3-4 scales) showed a ceiling effect (>25% at maximum score). Internal consistency: All Cronbach's alpha >0.70. Test-retest Intraclass Correlation Coefficients (ICCs) were moderate or adequate (>or=0.50; p < 0.01) for 10 ITQOL scales. VALIDITY: ITQOL-scales, with a few exceptions, correlated better with predefined parallel TAPQOL scales than with non-parallel scales. Five to eight ITQOL-scales discriminated clearly between children with few and with many parent reported chronic conditions, between children with and without doctor-diagnosed respiratory disease and with a low and a high parent-reported medical consumption (p < 0.05). CONCLUSIONS: This study supported the evidence that the ITQOL is a feasible instrument with adequate psychometric properties. The study provided reference ITQOL scores for gender/age subgroups. We recommend repeated evaluations of the ITQOL in varied populations, especially among very young children, including repeated assessments of test-retest characteristics and evaluations of responsiveness to change. We recommend developing and evaluating a shortened ITQOL version. PMID- 17111232 TI - Quality of life after radiofrequency ablation combined with transcatheter arterial chemoembolization for hepatocellular carcinoma: comparison with transcatheter arterial chemoembolization alone. AB - OBJECTIVE: Comprehensive studies are scarce with regard to the quality of life (QOL) of hepatocellular carcinoma (HCC) patients after transcatheter arterial chemoembolization (TACE) and/or radiofrequency ablation (RFA) treatment. The purpose of this study was to examine the impact of TACE alone and that of the TACE followed by RFA (TACE-RFA) on QOL in HCC patients. METHODS: QOL was measured using the Functional Assessment of Cancer Therapy-General (FACT-G) (Version 4.0) questionnaire, translated into Chinese, as a measure of QOL among Chinese HCC patients. Prospectively, 83 patients in both TACE group (n = 40) and TACE-RFA group (n = 43) completed the questionnaire at baseline and 3 months after treatment, respectively. RESULTS: The TACE-RFA group resulted in a significantly higher total QOL score, socio-family well-being score, and functional well-being score than that of TACE group, 3 months after respective treatment. Liver function, tumor recurrence and complication, age, income were the most important factors affecting the QOL of HCC patients after treatment. The logistic regression analyses showed that Child-Pugh Class and tumor recurrence after treatment were independent predictors of post-treatment QOL scores of HCC patients. CONCLUSIONS: The overall QOL of HCC patients in TACE-RFA group was maintained at a relatively higher level than that of TACE group. TACE followed by RFA appeared to be more favorable than TACE alone with respect to QOL. PMID- 17111233 TI - Rapid detection of differential item functioning in assessments of health-related quality of life: The Functional Assessment of Cancer Therapy. AB - REASON FOR STUDY: Differential item functioning (DIF) occurs when a test item functions differently in different groups when controlling for the level of the underlying construct measured by the test. DIF assessment is a first step in the evaluation of test bias. We sought to demonstrate a rapid hybrid approach to DIF detection by determining the presence and scale-level impact of DIF related to eight covariates in four domains measured by the Functional Assessment of Cancer Therapy (FACT). MAJOR FINDINGS: The number of items found with DIF in each domain depended on the criterion chosen to define the presence of DIF. With a few exceptions, scale-level differential functioning was similar regardless of the criteria chosen. For physical well-being, there was relevant scale-level differential functioning related only to race. For social and family well-being, there was relevant scale-level differential functioning related to each of the covariates. For emotional well-being, there was relevant scale-level differential functioning related to ethnicity, language, and race. For functional well-being, there was relevant scale-level differential functioning related to ethnicity, race, education, and self- vs. interviewer-administration. PRINCIPAL CONCLUSIONS: Our rapid hybrid approach to DIF detection may be broadly applicable in other studies of health-related quality of life. PMID- 17111234 TI - Globalization and fruitfly invasion and expansion: the medfly paradigm. AB - The phytophagous insects of the Tephritidae family commonly referred to as "true fruit flies" offer different case histories of successful invasions. Mankind has played an important role in altering the distributions of some of the more polyphagous and oligophagous species. However, the question arises why only a few species have become major invaders. The understanding of traits underlying adaptation in different environments is a major topic in invasion biology. Being generalists or specialists, along the K-r gradient of the growth curve, make a difference in term of food resources exploitation and interspecies competition and displacement. The species of the genus Ceratitis are good examples of r strategists. The genetic and biological data of the most notorious Ceratitis species, the Mediterranean fruit fly Ceratitis capitata (medfly), are reviewed to investigate the traits and behaviours that make the medfly an important invader. It can be learnt from medfly, that invasions in a modern global trade network tend to be due to multiple introductions. This fact allows a maintenance or enhancement of genetic variability in the adventive populations, which in turn increases their potential invasiveness. Our current knowledge of the medfly genome opens the way for future studies on functional genomics. PMID- 17111236 TI - Rapid light-response curves of chlorophyll fluorescence in microalgae: relationship to steady-state light curves and non-photochemical quenching in benthic diatom-dominated assemblages. AB - Rapid light-response curves (RLC) of variable chlorophyll fluorescence were measured on estuarine benthic microalgae with the purpose of characterising its response to changes in ambient light, and of investigating the relationship to steady-state light-response curves (LC). The response of RLCs to changes in ambient light (E, defined as the irradiance level to which a sample is acclimated to prior to the start of the RLC) was characterised by constructing light response curves for the RLC parameters alpha (RLC), the initial slope, ETR(m,RLC), the maximum relative electron transport rate, and E (k,RLC), the light-saturation parameter. Measurements were carried out on diatom-dominated suspensions of benthic microalgae and RLC and LC parameters were compared for a wide range of ambient light conditions, time of day, season and sample taxonomic composition. The photoresponse of RLC parameters was typically bi-phasic, consisting of an initial increase of all parameters under low ambient light (E < 21-181 micromol m(-2) s(-1)), and of a phase during which alpha (RLC) decreased significantly with E, and the increase of ETR(m,RLC) and E (k,RLC) was attenuated. The relationship between RLC and LC parameters was dependent on ambient irradiance, with significant correlations being found between alpha (RLC) and alpha, and between ETR(m,RLC) and ETR(m), for samples acclimated to low and to high ambient irradiances, respectively. The decline of alpha (RLC) under high light (Deltaalpha (RLC)) was strongly correlated (P < 0.001 in all cases) with the level of non-photochemical quenching (NPQ) measured before each RLC. These results indicate the possibility of using RLCs to characterise the steady-state photoacclimation status of a sample, by estimating the LC parameter E (k), and to trace short-term changes in NPQ levels without dark incubation. PMID- 17111235 TI - RhoGDIbeta lacking the N-terminal regulatory domain suppresses metastasis by promoting anoikis in v-src-transformed cells. AB - Rho guanine nucleotide dissociation inhibitors (RhoGDIs) regulate the activity of Rho family GTPases. RhoGDIbeta (LyGDI/GDID4/RhoGDI2) has two caspase cleavage sites after Asp19 and Asp55. The resulting cleavage products, DeltaN(1 19)RhoGDIbeta and DeltaN(1-55)RhoGDIbeta, are expressed in cells under conditions that activate caspases. DeltaN(1-19)RhoGDIbeta, which can inhibit GDP dissociation, is implicated in the process of apoptosis, whereas the physiological roles for DeltaN(1-55)RhoGDIbeta, which lacks the ability to inhibit GDP dissociation, are largely unknown. To explore the roles of DeltaN(1 55)RhoGDIbeta, we examined the phenotypes of v-src-transformed metastatic fibroblasts transfected with plasmids for expressing DeltaN(1-55)RhoGDIbeta. Although the expression of DeltaN(1-55)RhoGDIbeta had no effect on the rate of growth in vitro, it suppressed experimental metastasis and decreased the rate of growth in vivo. In addition, DeltaN(1-55)RhoGDIbeta-expressing cells had enhanced adhesion to fibronectin, laminin, and collagens but reduced retention in the lung after intravenous injection. Also, the expression of DeltaN(1-55)RhoGDIbeta promoted anoikis without affecting the levels of activated Rac1 or Cdc42. Furthermore, DeltaN(1-55)RhoGDIbeta did not affect the expression or phosphorylation of focal adhesion kinase, p44/p42 mitogen-activated protein kinases, or Akt1 before or after induction of anoikis. Thus, DeltaN(1 55)RhoGDIbeta appears to promote anoikis by undefined mechanisms, thereby suppressing metastasis in v-src-transformed fibroblasts. PMID- 17111237 TI - The atypical iron-coordination geometry of cytochrome f remains unchanged upon binding to plastocyanin, as inferred by XAS. AB - The transient complex between cytochrome f and plastocyanin from the cyanobacterium Nostoc sp. PCC 7119 has been analysed by X-ray Absorption Spectroscopy in solution, using both proteins in their oxidized and reduced states. Fe K-edge data mainly shows that the atypical metal coordination geometry of cytochrome f, in which the N-terminal amino acid acts as an axial ligand of the heme group, remains unaltered upon binding to its redox partner, plastocyanin. This fact suggests that cytochrome f provides a stable binding site for plastocyanin and minimizes the reorganization energy required in the transient complex formation, which could facilitate the electron transfer between the two redox partners. PMID- 17111238 TI - Self-assembled monolayer of light-harvesting core complexes of photosynthetic bacteria on an amino-terminated ITO electrode. AB - Light-harvesting antenna core (LH1-RC) complexes isolated from Rhodospirillum rubrum and Rhodopseudomonas palustris were successfully self-assembled on an ITO electrode modified with 3-aminopropyltriethoxysilane. Near infra-red (NIR) absorption, fluorescence, and IR spectra of these LH1-RC complexes indicated that these LH1-RC complexes on the electrode were stable on the electrode. An efficient energy transfer and photocurrent responses of these LH1-RC complexes on the electrode were observed upon illumination of the LH1 complex at 880 nm. PMID- 17111239 TI - Use of gastroprotective agents in recommended doses in hospitalized patients receiving NSAIDs: a drug utilization study. AB - OBJECTIVE: In recent years, studies investigated to what extend recommendations for co-prescribing gastroprotective agents in prevention of NSAID-induced gastrointestinal complications are followed in clinical practice. However, only a few studies have also taken into consideration the recommended dose of gastroprotectives prescribed in NSAID-induced ulcer prophylaxis. The aim of our study was to evaluate the prevalence of concomitant use of gastroprotectives with NSAIDs in hospitalized patients, with emphasis on the recommended dose of gastroprotectives for ulcer prophylaxis. METHOD: This observational, cross sectional, drug utilization study included all adult patients receiving NSAIDs hospitalized in the Clinical Hospital Center Zagreb on the day of the study. Data on age, sex, comorbidities, indications for NSAID use, type/dose of NSAIDs and gastroprotectives, history of gastrointestinal events, active gastrointestinal symptoms and risk factors were evaluated. MAIN OUTCOME MEASURE: Study outcomes were: (1) prevalence of prescription of gastroprotectives among NSAID-users at risk; (2) prevalence of prescription of gastroprotective in recommended dose; (3) association between risk factors and prescription of GPAs. RESULTS: The rates of gastroprotectives prescription were significantly higher in NSAID-users with concomitant risk factors as compared to patients without risk factors [47/70 (67.1%) and 8/22 (36.4%), respectively; p=0.01072]. However, gastroprotection in recommended ulcer-preventive dose was low in both groups [8/70 (11.4%) and 9/92 (9.8%), respectively]. The number of concomitant risk factors did not increase the odds of receiving anti-ulcer therapy (odds ratio 0.7279). Thirty-three percent of patients with concomitant risk factors were not prescribed gastroprotectives. Ibuprofen, NSAID with the lowest risk of inducing gastrointestinal complications, was prescribed in only two patients. CONCLUSION: The results indicate high awareness among hospital physicians about possible NSAID-induced gastrointestinal complications, but insufficient knowledge about risk factors related to NSAID-induced gastrointestinal toxicity, recommended dose of gastroprotectives in NSAID-induced ulcer prophylaxis and gastrointestinal toxicity of different types of NSAIDs. PMID- 17111240 TI - Development of a software tool for computation of parenteral nutrition in adults, and its potential role in improving nutritional care. AB - The detailed development of a computer program for management of parenteral nutrition (PN) in adults is described. Strengths and weaknesses are mentioned. By using the software, individualized PN prescriptions based on patient age, weight, sex, and clinical condition can be determined and available formulations can be automatically selected from incorporated databases. Automated PN management potentially enhances our ability to provide nutritional support with greater accuracy and precision and reduces the time required for writing and calculations. Day-to-day changes may also be monitored rapidly, simply, and safely. PMID- 17111241 TI - A 4-year study of lithium intoxication reported to the Czech Toxicological Information Centre. AB - OBJECTIVE: To evaluate the frequency and severity of lithium intoxication in calls to the Czech Toxicological Information Centre (TIC). METHOD: A 4-year retrospective study (2000-2003) of cases of lithium intoxication. Analysis of data from the database of the TIC and hospital discharge reports: sex, age, dose, blood level and biochemical markers of nephrotoxicity, symptoms, treatment and outcome of intoxication. RESULTS: The TIC received 70 calls concerning lithium intoxication, but only 27 discharge reports from hospitals were obtained and evaluated. Calls concerning women (16, median age 43.5 years) were more frequent than calls concerning men (11, median age 51.0 years). 16 patients had central nervous system or neurological symptoms. Signs of nephrotoxicity were present in 10 patients. Nine patients had cardiovascular symptoms. Possible interactions with other drugs during chronic overdoses were present in 14 patients. Six patients died due to lithium intoxication. CONCLUSION: Lithium intoxication remains a serious problem in calls to the TIC. Severe symptoms mainly developed in older patients. Drugs significantly potentiating toxic reactions to lithium should be avoided. PMID- 17111242 TI - Extreme thrombocytosis under the treatment by amoxicillin/clavulanate. AB - Amoxicillin/Clavulanate related extreme thrombocytosis during the treatment of pneumonia has never been reported. We present a 54-year-old patient who was admitted due to pneumonia, and was treated with amoxicillin/clavulanate IV and 3 day course of 500 mg azithromycin. Despite clinical and radiological evidence showing that pneumonia improved, thrombocytes increased rapidly. After stopping the antibiotic, the thrombocytes returned gradually to normal. Considering the clinical course, we propose that this extreme thrombocytosis was caused by the administration of amoxicillin/clavulanate. We describe this rare and unique patient and review the literature. PMID- 17111243 TI - Use and costs of anti-secretory and cardiovascular co-medication in osteoarthritis patients treated with selective or non-selective NSAIDS. AB - OBJECTIVE: This study aims to compare use and costs of anti-secretory and cardiovascular co-medication in osteoarthritis patients treated with selective or non-selective NSAIDs. METHOD: A retrospective study examined Belgian patients aged 65 years or more who suffer from osteoarthritis and are chronic users of selective NSAIDs (n=1,376) or non-selective NSAIDs (n=8,482). A before-and-after analysis compared drug use and costs between period 1 (first 6 months of 2002) and period 2 (several 1-year periods stretching over 2003-2004). A cohort analysis contrasted patients taking selective NSAIDs with patients taking non selective NSAIDs. MAIN OUTCOME MEASURES: Anti-secretory co-medication included histamine H2-receptor antagonists and proton pump inhibitors. Cardiovascular co medication referred to cardiac glycosides, anti-arrhythmics, anti-thrombotics, anti-angina drugs, anti-hypertensive drugs and serum-lipid-reducing drugs. Volume of drug use was expressed as number of packages and costs were computed in Euro. RESULTS: The volume of anti-secretory co-medication increased by 36% with selective NSAIDs and by 55% with non-selective NSAIDs between periods 1 and 2. Cardiovascular co-medication rose by 18% with selective NSAIDs and by 12% for non selective NSAIDs. Focusing on patients who did not take anti-secretory co medication in period 1, patients taking selective NSAIDs were just as likely to start anti-secretory co-medication in period 2 as patients taking non-selective NSAIDs (odds ratio: 1.05; 95% confidence interval: 0.90-1.23). Patients taking selective NSAIDs were just as likely to start cardiovascular co-medication as patients taking non-selective NSAIDs (odds ratio: 1.03; 95% confidence interval: 0.78-1.36). Annual costs of treating osteoarthritis in ambulatory care amounted to 756 with selective NSAIDs and 416 with non-selective NSAIDs. This originated from higher acquisition costs (278 vs. 24 ) and higher costs of co-medication (477 vs. 392 ) with selective NSAIDs. CONCLUSIONS: The use of selective and non-selective NSAIDs is accompanied by a higher use of co-medication over time. The increase in anti-secretory co medication was more prominent with non-selective NSAIDs. The rise in cardiovascular co-medication was more pronounced with selective NSAIDs. Treatment of osteoarthritis with selective NSAIDs is more expensive than with non-selective NSAIDs in terms of acquisition costs and costs of co-medication. PMID- 17111244 TI - Pharmaceutical care and its relationship to prescribing behaviour of general practitioners. AB - OBJECTIVE: To study the correlation between pharmaceutical care and prescribing routines of general practitioners (GPs). METHODS: Cross-sectional study; 201 pharmacies, 408 general practices, The Netherlands, 2000/2001. The variation in prescribing behaviour was characterised using 20 validated prescribing indicators based on general practice guidelines. The general construct 'adherence to guidelines' served as the dependent variable and was formed by summing the scores of the prescribing indicators. Four possible determinants of the variation were determined on the basis of survey questions: the construct 'the pharmacist's attitude towards pharmaceutical care', and three partial constructs derived from the pharmacist's care-providing function: the care for the individual patient, the cooperation with general practitioners and the registration of the care provided. A multiple linear regression analysis was then performed. MAIN OUTCOME MEASURE: The weighted score for the prescribing indicators. RESULTS: The weighted average score for the prescribing indicators was 65% (SD 3.7). The response rate to the survey was 71%. The pharmacist's attitude to pharmaceutical care, as well as the degree to which the pharmacist provided care for the individual patient, the degree to which he cooperated with the general practitioner and the degree to which he registered the care provided were not correlated with the 'adherence to guidelines' by the general practitioner with whom the pharmacist frequently cooperated. CONCLUSION: Variations between general practitioners in the quality of prescribing, as measured by their adherence to guidelines, were not correlated with pharmaceutical care by the pharmacist with whom they cooperated on a day-to day basis. PMID- 17111245 TI - Inappropriate drug use by Portuguese elderly outpatients--effect of the Beers criteria update. AB - OBJECTIVE: To characterize the use of medicines and to evaluate the inappropriateness of drugs in elderly outpatient population. SETTING: Twelve community pharmacies in different districts of Lisbon-Portugal. METHOD: Observational cross-sectional survey, in a sample of 213 elderly outpatients (age>or=65-years-old) presenting a prescription with two or more drugs, for their own use. MAIN OUTCOME MEASURES: Drug use pattern and prevalence of potentially inappropriate medication. RESULTS: We have studied 213 outpatients, who were taking a total of 1,543 drugs, with an average of 7.23 per patient. The drugs were distributed mainly in the following 3 ATC (Anatomical Therapeutic Chemical Classification) classes: C (cardiovascular system), N (nervous system) and A (alimentary tract). Using the 1997 Beers Explicit criteria, 75 occurrences of inappropriate medicines were detected in 59 patients (27.7%), while with the 2003 Beers Explicit criteria we detected 114 cases of inappropriate medication in 82 patients (38.5%). The occurrence of inappropriate medicines was significantly associated with the consumption of a high number of drugs. According to the ATC Classification, more than one half of the cases of inappropriateness were related with long acting benzodiazepines and with ticlopidine. The 2003 version detected a significantly higher prevalence of inappropriate drug use having potentially adverse outcomes of high severity. CONCLUSIONS: The application of the updated Beers criteria lead to higher rates of potentially inappropriate medication, and especially those responsible for more severe adverse outcomes. The results suggest that there is a need for interventions to improve instructions for safe drug use in the elderly patients and to decrease the number of medications whenever it is possible. This study suggests a high prevalence of potentially inappropriate drug use by the elderly patients of Lisbon region, Portugal. PMID- 17111246 TI - Evaluation of drotrecogin alpha use in a Belgian university hospital. AB - OBJECTIVE: Sepsis remains a major cause of mortality in ICU patients, despite advances in therapy. Drotrecogin alpha (Drot AA), a recombinant human activated protein C with anti-inflammatory and anticoagulant properties, has proven to be successful in patients with at least one organ failure. Our goal was to evaluate the data in patients with sepsis and at least two organ dysfunctions in a large university hospital in Belgium. SETTING: The study was conducted at the medical and surgical intensive care units of the 1850-bed university hospital of Leuven, Belgium. METHODS: We retrospectively evaluated the use of Drot AA during a 2.5 year period. At baseline patients' demographics, type of infection, APACHE II (acute physiology and chronic health evaluation), SOFA (sequential organ failure assessment), DIC (diffuse intravascular coagulation) score and number of organ failures were obtained. Overall hospital mortality was defined as primary outcome measure. Special attention was paid to bleeding, the main side effect of Drot AA. MAIN OUTCOME MEASURE: Evalution of hospital and ICU mortality in patients treated with Drot AA for severe sepsis. RESULTS: Drot AA was administered to 23 patients with sepsis and at least 2 organ dysfunctions; all patients started treatment within 24 h of onset of the second organ failure. Mean age was 59 years. Mean number of organ failures was 3. Overall hospital mortality rate was 47.8%. A 28 day mortality of 26% was found, comparable with the 28-day mortality rate of the PROWESS trial. Bleeding, requiring more than 3 units of blood, occurred in 1 patient. Although underlying co-morbidity was more pronounced in survivors, non survivors had a slightly higher median APACHE II, higher SOFA score and higher DIC score. However, the number of organ failures was identical in both groups. CONCLUSIONS: Overall hospital mortality rate was similar as observed in the Belgian Registry and 28-day mortality was equal to the results of the PROWESS study. Due to the limited number of patients, it is not clear if patients should be selected based on APACHE II, DIC or number of organ failures. However, selection based on number of organ failures is more appropriate due to intrinsic problems of the APACHE II score. PMID- 17111247 TI - Patients' attitudes towards and experiences of generic drug substitution in Norway. AB - OBJECTIVE: The aim of the study was to assess patients' attitudes towards and experiences of generic substitution 3 years after generic substitution of prescription medicines was permitted in Norway. METHODS: Prescriptions from 2,128 consecutive patients in a Norwegian pharmacy were retrospectively reviewed to identify all patients (n=274) receiving eight or more different prescription drugs on the fifth level in the Anatomical Therapeutic Chemical (ATC) classification system during the last 6 months. An age-adjusted control group (n=269) of patients receiving three to seven different prescription drugs was randomly selected. Of the 543 patients, 386 were eligible for inclusion. Both groups received a mailed questionnaire addressing their experiences with and attitudes towards generic substitution. RESULTS: The overall response rate was 73% (281/386) and the average age of the respondents was 66 years old. The study found that patients who reported to have received information from their physician or the pharmacy about generic substitution were more likely to have switched (P<0.001). About half (138/281) of the patients had experienced a generic switch, and a higher proportion of the polypharmacy patients had their medication substituted compared to the control patients (P<0.001). Altogether 50 (36%) of the patients who had experienced a switch, reported one or more negative experiences connected to the substitution, and 29 of these (21%) reported an overall negative experience after the change. The experiences of the patients were not related to age, gender, or number of medications or information about generics from either the pharmacy or the physician. About 41% of the patients would not switch if they had no personal economic incentives. CONCLUSIONS: About 1/3 of the patients who had their medication substituted reported negative experiences. Generic drug substitution for a number of patients is not considered an equal alternative to branded drugs, and these patients may need additional information and support. The lack of correlation between patient experiences and age, gender, and medical regimen complexity is interesting and should be investigated further. PMID- 17111248 TI - Science or practice? UK undergraduate experiences and attitudes to the MPharm degree. AB - OBJECTIVE: The debate surrounding the science/practice balance in the teaching of undergraduate pharmacy has been played out in the professional literature for years. The objective of this work was to explore the attitudes of pharmacy undergraduates on the practice-science debate. SETTING: The study was undertaken as part of a national study of teaching, learning and assessment methods in United Kingdom (UK) schools of pharmacy. METHOD: Six focus groups were carried out. The sample was 44 volunteer students from nine UK schools of pharmacy, representing all 4 years of the MPharm programme. Groups were tape recorded and transcribed. Analysis of the transcripts was theme based by topic. MAIN OUTCOME MEASURE: Qualitative data on student attitudes and experiences. RESULTS: Most students thought that there was too strong an emphasis placed on the science components of the course in the early part of their studies. Later in the course they realised that the majority of the science was necessary; it just had not been apparent to them at the time. There were strongly held attitudes across all 4 years that it would be beneficial to include more practice-related material at the beginning of their studies. This would be beneficial for three reasons: to make the course more interesting, to aid in the contextualisation of the science component and to assist the students in any early placement or vacational work. CONCLUSION: Internationally, changes to the role of the pharmacist from a traditional supply function to a more clinical role has resulted in differing educational needs for the pharmacist of the future. Pharmacy will remain a degree built on a strong scientific background, but students advise that the contextualisation and sequencing of material within the degree could make a considerable improvement to their learning. Consulting students helps us to understand the teaching, learning and assessment experience better by giving insights into ways of improving the delivery. In the case of the UK, there are legislative changes impending which may provide an opportunity to review the balance of practice-and science in the curriculum. PMID- 17111249 TI - Concurrent and separate effects of body mass index and waist-to-hip ratio on 24 year mortality in the Population Study of Women in Gothenburg: evidence of age dependency. AB - Obesity is generally assumed to be an important risk factor for death and morbidity. However, the association between excess body weight and all-cause mortality among younger and older women and the impact of body mass index (BMI) and waist-to-hip ratio (WHR) concurrently is not fully understood. In 1968-1969 we initiated a prospective study comprising a population sample of 1,462 women from Gothenburg, Sweden. During a 24 year period, until 1992-1993, 265 women had died. A multivariable Cox Proportional Hazards Regression model was used to estimate the relative risk of death in relation to BMI and WHR, with age and other covariates of age-specific interest as smoking, physical activity at work and leisure time and serum triglyceride concentration, at start of the study. BMI and WHR were analyzed as independent variables. Younger women (38 and 46 years at baseline) presented a statistically significant non-linear (U-shaped) relation between BMI and mortality. Among older women (50, 54 and 60 years at baseline), a significant negative linear relationship with decreasing mortality in relation to increasing BMI values was seen. For all women a higher WHR was related to an increased risk of death. The lowest risk of death among younger women corresponded to a low WHR and a BMI within the middle range. For older women the highest survival was observed for those with lowest WHR and highest BMI. Thus, in older women a high BMI seems not to be an increased risk as long as adiposity is not centrally located. PMID- 17111250 TI - The first database comprised of flagellin gene (flaA) types of Campylobacter jejuni human clinical isolates from Greece. AB - BACKGROUND: Flagellin subunit A gene (flaA) typing of Campylobacter has been recognized by several groups as a relatively simple and quick genotyping method. The present study aimed to create, for the first time in Greece, a database with flaA restriction patterns, which could be used for future epidemiological and clinical studies. A total of 207 C. jejuni clinical isolates of known serotype were collected from 5 general hospitals of the area of Attica, during the period 2000-2003. RESULTS: The RFLP profiles of each strain were matched in 44 bins of 0 or 1. Thirty nine different flaA types, designated as flaA 1 GR to flaA 39 GR (GR: Greece) were found. There was no significant association of certain genotypes with certain serotypes. However flaA typing showed a remarkable discriminatory ability inside the non-typable (NT) group. CONCLUSIONS: Evaluating our results we observed (i) that there was no clonality of a certain flaA type among the strains and the serotypes examined and (ii) that the discriminatory ability of flaA typing was much better than that of serotyping. Giving a simple and detailed description of the data analysis, we are the first who publish the bin patterns for the flaA genotypes found. PMID- 17111251 TI - Duration of pregnancy in relation to seafood intake during early and mid pregnancy: prospective cohort. AB - We examined the association between exposure to seafood intake during two periods of pregnancy on the one hand and risks of preterm delivery and postterm delivery on the other. In a prospective cohort of 8729 pregnant Danish women, we assessed frequency of fish meals during the first and second trimester of pregnancy by questionnaires completed around gestation weeks 16 and 30, respectively. When fish intake was based solely on intake reported for the early period of pregnancy, mean gestation length was shorter by 3.91 (95% CI: 2.24-5.58) days and odds of preterm delivery were increased 2.38 (1.23-4.61) times in those who never consumed fish (n = 308) vs. those who consumed both fish as main meal and fish in sandwiches at least once per week (n = 785). These measures were similar when fish intake was based solely on intake reported for mid-pregnancy. In the subgroup of women reporting same intake in the two trimesters, those who never consumed fish (n = 165) had 8.57 (5.46-11.7) days shorter mean gestation and 19.6 (2.32-165) times increased odds of preterm delivery, compared to high fish consumers (n = 127); odds of elective and postterm delivery were reduced by a factor 0.33 (0.11-1.02) and 0.34 (0.12-0.95), respectively, in zero fish consumers. All analyses were adjusted for potential confounding by factors such as maternal smoking, height, and prepregnant weight. We conclude that never consuming fish in the first two trimesters of pregnancy was an extremely strong risk factor for preterm delivery but was also associated with reduced risks of elective delivery and postterm delivery. PMID- 17111252 TI - Projecting the number of patients with colorectal carcinoma by phases of care in the US: 2000-2020. AB - OBJECTIVE: This study provides projections of colorectal cancer prevalence by phases of care (initial, monitoring, and last year of life) to the year 2020 and describes the estimation method. METHODS: Cancer prevalence by phase of care was estimated from colorectal cancer incidence and survival from the Surveillance, Epidemiology, and End Results (SEER) Program data, population estimates and projections from the US Census Bureau, and all cause mortality data from the Human Mortality Life Tables. Assumptions of constant incidence and survival were used for projections from 2000 to 2020. Modeled and directly observed patient months by phase of care were compared for 1996 -1998 to provide validation of estimates. RESULTS: Prevalence of colorectal cancer is estimated to increase from 1,002,786 (0.36%) patients to 1,522,348 (0.46%) patients between 2000 and 2020. The estimated number of person-months in the initial and last year of life phases of care will increase 43%, while the monitoring phase of care will increase 54%. Modeled person-months by phase of care were consistent with directly observed measures of person months by phase of care in 1996-1998. CONCLUSIONS: Under assumptions of current cancer control strategies we project that colorectal cancer prevalence will increase more rapidly than the US population, largely due to the aging of the US population. This suggests that considerable resources will be needed in the future for initial, continuing and last year of life treatment of colorectal cancer patients unless notable breakthroughs in primary prevention occur in the future years. PMID- 17111253 TI - Do breast cancer risk factors modify the association between hormone therapy and mammographic breast density? (United States). AB - OBJECTIVE: To evaluate whether the association between hormone therapy (HT) and breast density differs by levels of breast cancer risk factors. METHODS: We evaluated 80,867 screening mammograms from 39,296 postmenopausal women from Washington State. We estimated odds ratios and 95% confidence intervals for dense breasts (Breast Imaging Reporting and Data System categories 3 "heterogeneously dense" and 4 "extremely dense") compared to fatty breasts (categories 1 "almost entirely fat" and 2 "scattered fibroglandular") among HT users compared to never users. We separately examined former HT use and current HT use by type (estrogen plus progestin therapy (EPT) and estrogen-only therapy (ET)). We stratified the associations by age, BMI, race, family history, and reproductive and menopausal factors. RESULTS: Current EPT users had a 98% (1.87-2.09) greater odds of having dense breasts and current ET users had a 71% (1.56-1.87) greater odds compared to never users. Current HT users were more likely to have dense breasts if they were older, had more children, or younger at first birth compared to never users; these associations were stronger among EPT users than ET users. CONCLUSIONS: HT, particularly EPT, may reduce protective effects of older age, parity, and younger age at first birth on mammographic density. PMID- 17111254 TI - Sex steroid hormones in young manhood and the risk of subsequent prostate cancer: a longitudinal study in African-Americans and Caucasians (United States). AB - OBJECTIVE: To investigate the relation of sex hormone levels in young adults to subsequent prostate cancer risk. METHODS: From 1959 to 1967, the Child Health and Development Studies collected sera from 10,442 men (median age: 34 years) and followed them for a median of 32 years. In this analysis, we selected 119 African Americans and 206 Caucasians diagnosed with prostate cancer during the follow-up period. Two prostate cancer-free men were chosen to match each prostate cancer case on race and birth year. We compared the levels of testosterone, estradiol, and sex hormone-binding globulin in cases to those of their matched controls using conditional logistic regression. RESULTS: There was no significant association between absolute levels of sex hormones in youth and prostate cancer risk in either race. However, among Caucasians, but not African-Americans, prostate cancer risk was positively associated with the ratio of total testosterone to total estradiol (odds ratio relating the fourth to the first quartile: 3.01; 95% confidence interval: 1.42-6.39). CONCLUSIONS: The association between testosterone to estradiol ratio and prostate cancer risk in young Caucasians is consistent with similar findings in older Caucasians. The absence of this association in African-Americans needs confirmation in other data involving larger numbers of African-Americans. PMID- 17111255 TI - Serum triglycerides and colorectal adenoma in a case-control study among cancer screening examinees (Japan). AB - OBJECTIVE: Most epidemiologic studies have shown serum triglycerides to be associated with colorectal adenoma. However, whether the association can be modified by smoking is unknown. We cross-sectionally investigated the association of serum triglycerides with the risk of adenoma by smoking status. METHODS: We identified 782 newly diagnosed adenoma cases from the examinees of a colorectal cancer screening program. All cases were diagnosed by a magnifying colonoscopy with dye spreading. We determined 738 controls without present illness or past history of adenoma from among the examinees. They provided their lifestyle information and fasting blood samples to measure their serum triglycerides. We calculated odds ratios (OR) and 95% confidence intervals (CI) of colorectal adenoma for serum triglycerides. RESULTS: High serum triglycerides were associated with colorectal adenoma (OR 1.5; 95% CI 1.1-2.0 for the highest versus the lowest quartile, P (trend, )0.030). A stronger association was observed between three or more adenoma cases and study controls (OR 2.3; 95% CI 1.3-4.2, P (trend,) < 0.0010). After classifying the study subjects by smoking status, a significant linear risk trend was found in ever-smokers (P (trend), 0.0018) but not in never-smokers (P (trend), 0.94; P (interaction), 0.067). CONCLUSIONS: Our results suggested that a higher serum triglyceride level may be related to a larger number of adenomas. Adenoma development involving an elevated serum triglyceride level may be modified by smoking. PMID- 17111256 TI - Adolescent dietary phytoestrogen intake and breast cancer risk (Canada). AB - OBJECTIVE: It has been suggested that dietary phytoestrogen intake during adolescence may reduce the risk of developing breast cancer. This population based case-control study evaluated the association between adolescent dietary phytoestrogen intake and adult breast cancer risk among women in Ontario, Canada. METHODS: Pathology-confirmed, population-based breast cancer cases, aged 25-74 years, diagnosed between June 2002 and April 2003, were identified using the Ontario Cancer Registry. Population-based controls were recruited, and matched to cases within 5-year age groups. Adolescent phytoestrogen intake was obtained using a brief food frequency questionnaire (n = 3,024 cases, n = 3,420 controls). Logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Higher phytoestrogen intake (both isoflavones and lignans) during adolescence was associated with a reduced breast cancer risk, and a monotonic trend was observed from the lowest to the highest quartile (OR [Q2] = 0.91, 95% CI 0.79-1.04, OR[Q3] = 0.86, 95% CI 0.75-0.98, and OR[Q4] = 0.71, 95% CI 0.62-0.82, p-trend < 0.001). CONCLUSION: Adolescent dietary phytoestrogen intake may be associated with a decreased risk of adult breast cancer. If verified, this finding has important implications with regard to breast cancer prevention since diet is a potentially modifiable factor. PMID- 17111257 TI - Hypothesis: hair cover can protect against invasive melanoma on the head and neck (Australia). AB - The anatomic distribution of cutaneous melanoma reflects people's levels and patterns of sun exposure. While examining trends of incident invasive melanomas by site in recent decades in Australia we noted significant increases in incidence on the ears but not the face or any other site in women younger than 40 years, by 6% (95% confidence interval [CI] 2-10%) per year, and 40-59 years by 7% (95% CI 4-10%) per year. Men of the same age showed no corresponding changes in ear melanoma. However incidence rates of ear melanoma in general were fourfold higher in males than females in Australia. Further, using data from the National Cancer Institute's Surveillance, Epidemiology and End Results (SEER) Program, rates of invasive melanoma on the ear were found to be sevenfold higher in males than females in the US population in the same period. Higher rates of scalp and neck melanomas were also seen in men and women in both populations. We therefore speculated that the isolated rises of ear melanoma in younger women in Australia, and the higher overall rates of ear, scalp and neck melanoma in men compared with women, reflect differences in hair coverage. We tested the specific hypothesis that hair cover reduces sun exposure of the ears using experimental manikins and found that hair cover of the ear reduced solar ultraviolet-B exposure by 81% [SE +/-8] compared with uncovered ears. We conclude that hair cover can protect against invasive melanoma on the ear and may similarly protect on the scalp and neck. When discretionary, hair may be an important additional factor to be considered for melanoma prevention. PMID- 17111258 TI - Early infection and risk of childhood brain tumors (Canada). AB - OBJECTIVE: Up to 15% of human cancers can be attributed to infections. Currently there are no known associations between infective agents and childhood brain tumors. We explored childhood brain tumor risk associated with a variety of indicators of infection during gestation and childhood. METHODS: Two hundred and seventy-two cases of childhood brain tumor diagnosed in children less then 15 years of age in the province of Quebec between 1980 and 1999 were included in the study. An equal number of sex and age matched population based controls were recruited from family allowance or provincial health insurance files. Conditional logistic regression was used to estimate the risk of developing childhood brain tumors associated with self-reported exposure to infection. RESULTS: Childhood brain tumor risk was weakly to moderately elevated after maternal reported exposure to several indicators of infection: use of antibiotics during gestation (OR = 1.7, 95% CI = 0.8-3.6) or childhood (OR = 1.4, 95% CI = 0.7-2.9), removal of tonsils, adenoids or appendix (OR = 1.2, 95% CI = 0.6-2.4), having siblings (OR = 1.4, 95% CI = 0.9-2.3), and being at least second born (OR = 1.7, 95% CI = 1.2-2.4). Moreover, childhood brain tumor risk was reduced for some subjects who were breastfed or attended daycare for more than 1 year. Risk varied by sex, age at diagnosis and tumor type. CONCLUSION: Childhood brain tumor risk may be associated with exposure to infective agents. PMID- 17111259 TI - Induced abortions and the risk of all cancers combined and site-specific cancers in Shanghai. AB - Although some previous case-control studies found an increased risk of breast cancer in women who had an induced abortion, the evidence from prospective studies suggests that induced abortions do not cause breast cancer. We have assessed risks of 12 types of cancer in women who have had induced abortions in a prospective study in China. Female textile workers (n = 267,400) completed a baseline questionnaire (1989-1991) that ascertained information on the major risk factors for breast cancer, contraceptive use, and induced abortions and were actively followed until July 2000. Cox Proportional Hazards analysis was used to calculate incidence rate ratios for specific types of cancer in women who ever had an induced abortion and by number of induced abortions. Women who had had an abortion were not at increased risk of cancer. There was a significant reduction in risk of uterine corpus cancer in women who had ever had an induced abortion, and a significant decreasing trend in risk with increasing number of induced abortions. No convincing associations with other cancers were observed. Women who have induced abortions after a live birth are not at increased risk of cancer and induced abortions may reduce risk of cancer of the corpus uteri. PMID- 17111260 TI - Breast cancer risk factors in relation to breast density (United States). AB - OBJECTIVES: Evaluate known breast cancer risk factors in relation to breast density. METHODS: We examined factors in relation to breast density in 144,018 New Hampshire (NH) women with at least one mammogram recorded in a statewide mammography registry. Mammographic breast density was measured by radiologists using the BI-RADS classification; risk factors of interest were obtained from patient intake forms and questionnaires. RESULTS: Initial analyses showed a strong inverse influence of age and body mass index (BMI) on breast density. In addition, women with late age at menarche, late age at first birth, premenopausal women, and those currently using hormone therapy (HT) tended to have higher breast density, while those with greater parity tended to have less dense breasts. Analyses stratified on age and BMI suggested interactions, which were formally assessed in a multivariable model. The impact of current HT use, relative to nonuse, differed across age groups, with an inverse association in younger women, and a positive association in older women (p < 0.0001 for the interaction). The positive effects of age at menarche and age at first birth, and the inverse influence of parity were less apparent in women with low BMI than in those with high BMI (p = 0.04, p < 0.0001 and p = 0.01, respectively, for the interactions). We also noted stronger positive effects for age at first birth in postmenopausal women (p = 0.004 for the interaction). The multivariable model indicated a slight positive influence of family history of breast cancer. CONCLUSIONS: The influence of age at menarche and reproductive factors on breast density is less evident in women with high BMI. Density is reduced in young women using HT, but increased in HT users of age 50 or more. PMID- 17111261 TI - Body size and composition and risk of rectal cancer (Australia). AB - BACKGROUND: Although body mass index has been shown to be associated with colon cancer, studies of rectal cancer risk have generally reported no association. The relationship between rectal cancer risk and central adiposity, overall fat mass, and fat-free mass is unknown. METHODS: In a prospective cohort study of people aged 27-75 years, body measurements were taken directly; fat mass and fat-free mass being estimated by bioelectrical impedance analysis and central adiposity by waist circumference and waist-to-hip ratio. Among 16,867 men and 24,247 women followed on average for 10.3 years, 229 rectal cancers were ascertained via the population cancer registry. RESULTS: When comparing the highest tertile with the lowest tertile, weight (hazard ratio = 1.4, 95% confidence interval (CI) 1.1 2.0), waist circumference (hazard ratio = 1.4, 95% CI 1.0-1.9), fat mass (hazard ratio = 1.4, 95% CI 1.0-2.0) and percent fat (hazard ratio = 1.4, 95% CI 1.0-2.0) were positively associated with rectal cancer risk. There was no evidence that risk differed by sex for any of the anthropometric measures. CONCLUSIONS: Waist circumference and fat mass may be weakly related to risk of rectal cancer. PMID- 17111262 TI - Aspirin may be more effective in preventing colorectal adenomas in patients with higher BMI (United States). AB - Obesity is a risk factor for colon cancer, possibly due to elevated levels of circulating cytokines derived from adipose tissue. Aspirin, which may affect the levels of these cytokines, has been shown in randomized controlled trials to decrease the risk of colorectal adenomas. We hypothesized that the chemopreventive effect of aspirin might be greater in individuals with higher body mass index (BMI). Data were available from the Aspirin/Folate Polyp Prevention Study, a randomized controlled trial of aspirin and folic acid to prevent recurrent colorectal adenomas. Obesity was defined as BMI > or = 30 (kg/m2), overweight as BMI of 25-29 (kg/m2) and normal weight as BMI <25 (kg/m2). For the analysis of the effect of aspirin on the recurrence of colorectal adenoma by BMI, we computed risk ratios for aspirin versus placebo within the three BMI strata using a modified Poisson model. Overall the risk reduction of adenomas with a daily dose of 325 mg aspirin was greater among subjects with higher BMI. Among obese subjects the risk ratio (RR) for advanced adenomas compared with placebo was 0.44 (95% CI 0.17-1.10), versus RR = 1.23 (95% CI 0.55-2.77) among those with normal weight. However, 81 mg aspirin daily did not interact with BMI to modify the risk of adenomas in such a fashion. The more pronounced effect of 325 mg aspirin in individuals with higher BMI suggests a possible protective role of anti-inflammatory aspirin against increased adipose-driven cytokines among obese subjects. PMID- 17111263 TI - The effect of occasional smoking on smoking-related cancers: in the European Prospective Investigation into Cancer and Nutrition (EPIC). AB - OBJECTIVE: Most studies on tobacco smoking have focused on daily-smokers. Occasional smokers, who have never smoked daily, have often been included in the reference group of never-smokers. We have investigated the association between occasional smoking and cancer of the bladder, kidney, pancreas, upper aero digestive tract and lung. METHODS: The study population consisted of 158,488 persons, who provided information on occasional smoking, within the European Prospective Investigation into Cancer and Nutrition (EPIC), 780 of whom developed a smoking-related cancer. We used Cox proportional hazard model, stratified by gender and country to estimate incidence rate ratios (IRR) for smoking-related cancers. RESULTS: The results suggest that occasional smokers have a higher risk of bladder cancer (IRR: 1.92, 95% confidence interval (CI) 0.93-3.98) and of the major smoking-related cancers combined (IRR: 1.24, 95% CI 0.80-1.94) than true never-smokers. Including occasional smokers in the reference group resulted in a lower risk estimate for former and current smokers. CONCLUSIONS: Occasional smoking should be discouraged. PMID- 17111266 TI - Developmental dyslexia: an update. AB - Dyslexia is the most common and carefully studied of the learning disabilities in school-age children. It is characterized by a marked impairment in the development of reading skills, and affects a large number of people (5-10%). Reading difficulties may also arise from poor vision, emotional problems, decreased hearing ability, and behavioral disorders, such as attention-deficit hyperactivity (ADHD). Although many areas of the brain are involved in reading, analysis of postmortem brain specimens by a variety of imaging techniques most consistently suggests that deficiency within a specific component of the language system - the phonologic module - in the temporo-parietal-occipital brain region underlies dyslexia. It is a highly familial and heritable disorder with susceptibility loci on chromosomes 1, 2, 3, 6, 11, 13, 15 and 18. Recently, four candidate genes (KIAA 0319, DYX1C1, DCDC2 and ROBO1) are shown to be associated with dyslexia. Although some of these results are controversial because of the genetic heterogeneity of the disorder, the available evidence suggests that dyslexia could be due to the abnormal migration and maturation of neurons during early development. Interestingly, in spite of genetic heterogeneity, the pathology appears to involve common phonological coding deficits. The condition can be managed by a highly structured educational training exercise. PMID- 17111267 TI - Human organic cation transporter (OCT1 and OCT2) gene polymorphisms and therapeutic effects of metformin. AB - Organic cation transporters (OCTs) are responsible for the hepatic and renal transport of metformin. In this study we analyzed variants of OCT1 and OCT2 genes in 33 patients (24 responders and nine non-responders) based on the hypothesis that polymorphisms in both genes contribute to large inter-patient variability in the clinical efficacy of metformin. The sequences of the 5'-flanking and coding regions of the two genes of interest were screened by single-strand conformation polymorphism (SSCP) analysis. To compare the causative factors between responders and non-responders, we performed stepwise discriminant functional analysis. Age, body mass index (BMI) and treatment with lipid-lowering agents were demonstrated as positive predictors, and two mutations in the OCT1 gene, -43T > G in intron 1 and 408Met > Val (1222A > G) in exon 7, were negative and positive predictors, respectively, for the efficacy of metformin; the predictive accuracy was 55.5% (P < 0.05). Subsequent study indicated that OCT1 mRNA levels tended to be lower in human livers with the 408Met (1222A) variant, though the differences did not reach the level of significance. In this study it is suggested that OCT1 and OCT2 gene polymorphisms have little contribution to the clinical efficacy of metformin. PMID- 17111268 TI - Flexible ex vivo phantoms for validation of diffusion tensor tractography on a clinical scanner. AB - PURPOSE: The aim of this study was to develop ex vivo diffusion tensor (DT) flexible phantoms. MATERIALS AND METHODS: Materials were bundles of textile threads of cotton, monofilament nylon, rayon, and polyester bunched with spiral wrapping bands and immersed in water. DT images were acquired on a 1.5-Tesla clinical magnetic resonance scanner using echo planar imaging sequences with 15 motion probing gradient directions. DT tractography with seeding and a line tracking method was carried out by software originally developed on a PC-based workstation. RESULTS: We observed relatively high fractional anisotropy on the polyester phantom and were able to reconstruct tractography. Straight tracts along the bundle were displayed when it was arranged linearly. It was easy to bend arcuately or bifurcate at one end; and tracts followed the course of the bundle, whether it was curved or branched and had good agreement with direct visual observation. Tractography with the other fibers was unsuccessful. CONCLUSION: The polyester phantom revealed a diffusion anisotropic structure according to its shape and would be utilizable repeatedly under the same conditions, differently from living central neuronal system. It would be useful to validate DT sequences and to optimize an algorithm or parameters of DT tractography software. Additionally, the flexibility of the phantom would enable us to model human axonal projections. PMID- 17111269 TI - Interfractional fluctuation of rectal dose in high dose rate brachytherapy for prostate cancer. AB - PURPOSE: The aim of the study was to explore the cause of the difference in the maximal rectal dose between the first and second high dose rate (HDR) brachytherapy applications by comparing the thickness of the anterior rectal wall. MATERIALS AND METHODS: The rectal dose and the thickness of the anterior rectal wall were analyzed in 26 patients with prostate cancer. After undergoing external beam radiation treatment with a total isocenter dose of 50 Gy, they were treated with HDR brachytherapy of 7.5 Gy/fraction, two fractions daily. The interval between the first HDR brachytherapy session and the second was 5 h. The rectal doses were directly surveyed during irradiation of the HDR brachytherapy. Thickening of the anterior rectal wall was measured at the same level by axial computed tomography scans obtained before the first and second HDR brachytherapy applications. RESULTS: The maximal surveyed rectal doses during the first and second HDR brachytherapy applications were 188 +/- 51 cGy and 220 +/- 35 cGy, respectively (P < 0.01). The fluctuation ratio exceeded 1 in each case. The thickness of the anterior rectal wall before the first and second HDR brachytherapy applications was 18.78 +/- 4.34 mm and 14.95 +/- 4.09 mm (P < 0.01), respectively. The fluctuation difference exceeded 0 in each case. CONCLUSION: The different rectal dose is attributable to thinning of the anterior rectal wall. The total rectal dose is within the range of doses at risk of exerting a toxic effect on the rectum. PMID- 17111270 TI - Evaluation of renal arteries in living renal donors: comparison between MDCT angiography and gadolinium-enhanced 3D MR angiography. AB - PURPOSE: The purpose of this study was to clarify and compare the accuracy of contrast-enhanced computed tomography (CT) angiography using multidetector-row helical CT (MDCT angiography) and gadolinium-enhanced MR angiography using three dimensional Fourier transformation gradient-echo sequence (3D MR angiography) for preoperative evaluation of renal arteries in living renal donors. MATERIALS AND METHODS: A total of 42 living renal donor candidates underwent both MDCT angiography and 3D MR angiography before digital subtraction angiography (DSA). Each MDCT angiogram and 3D MR angiogram was prospectively interpreted, and the findings were compared with the DSA results. RESULTS: MDCT angiography identified all of the 12 supernumerary arteries detected by DSA, whereas 3D MR angiography identified only 8. MDCT angiography identified all of the 19 proximal arterial branches detected by DSA, whereas 3D MR angiography identified only 16. CONCLUSION: A more accurate depiction of renal arteries in living renal donors can be achieved with MDCT angiography than with 3D MR angiography. PMID- 17111271 TI - Outcomes in 24 selected patients with stage IVB cervical cancer and excellent performance status treated with radiotherapy and chemotherapy. AB - PURPOSE: We sought to review outcomes in patients with stage IVB carcinoma of the cervix treated with irradiation in combination with chemotherapy. MATERIALS AND METHODS: We report outcomes of 24 consecutive patients with good performance status treated from 1998 to 2005. Most of these patients underwent concurrent irradiation with platinum-based chemotherapy. Some patients received subsequent systemic chemotherapy. RESULTS: All patients underwent external beam radiotherapy; 7 patients (29%) had additional high-dose-rate and 12 (50%) low dose-rate brachytherapy. Two patients (8%) received an IMRT boost instead of brachytherapy. The mean dose to point A was variable (73.9 +/- 19.2 Gy). Twenty patients (83%) received radio-sensitizing platinum-based chemotherapy, and the remaining had radiotherapy alone. Seven patients (29%) had further combination chemotherapy. Therapy was well tolerated. The overall survival was 44% at 36 months and 22% at 5 years. CONCLUSION: Patients with stage IVB cervical cancer have mostly been treated with palliative intent. With the advent of concurrent chemoradiation, we have treated many of these cases with aggressive combination therapy. In this series, the use of radiotherapy and multiagent chemotherapy in patients with stage IVB cervical carcinoma and good performance status was well tolerated and resulted in higher survival rates than previously reported. PMID- 17111272 TI - Brown tumor with fluid-fluid levels in a patient with primary hyperparathyroidism: radiological findings. AB - We report a case of a brown tumor with fluid-fluid levels in a patient with primary hyperparathyroidism. A 19-year-old woman presented with a 3-month history of pain in the left pubic region. The laboratory data showed elevated serum calcium and intact parathyroid hormone, confirming the diagnosis of primary hyperparathyroidism. Plain radiography and computed tomography (CT) showed an expansile lytic lesion of the superior ramus of the left pubis. The cortex was thinned. On magnetic resonance (MR) images, the lesion was solid and cystic. The solid area of the lesion showed heterogeneous low to intermediate signal intensity on T1-weighted images and heterogeneous low to high signal intensity on T2-weighted images. The cystic area showed several fluid-fluid levels on T2 weighted images. Dynamic contrast-enhanced MR images after administration of Gd DTPA showed marked, early enhancement of the solid area of the lesion. A delayed image showed prolonged enhancement of the solid area and enhancement of the septa and walls of the cystic area. Histopathology of a biopsy specimen showed fibroblastic proliferation, abundant giant cells, and focal hemosiderin deposition, which supported the diagnosis of a brown tumor. After removing the parathyroid adenoma, the brown tumor regressed and became sclerotic on radiographs. PMID- 17111273 TI - Acute dural venous sinus thrombosis without brain parenchymal abnormality: assessment with cerebral blood volume using dynamic susceptibility contrast magnetic resonance imaging. AB - We report results applying the dynamic susceptibility contrast (DSC) magnetic resonance (MR) technique to a patient with dural venous sinus thrombosis (DVST) of the right transverse-sigmoid sinus without brain parenchymal abnormality. The DSC-MR technique clearly demonstrated increased regional cerebral blood volume of the right temporo-parieto-occipital region adjacent to a right transverse-sigmoid sinus thrombosis in a patient with DVST without cerebral edema or hemorrhage. PMID- 17111274 TI - Squamous cell carcinoma arising from Stensen's duct. AB - We report a case of squamous cell carcinoma presumably arising from the left Stensen's duct. The tumor was discovered during management of recurrent left parotitis. Ultrasonography, computed tomography, and magnetic resonance imaging were useful for estimating the tumor and coexisting obstructive parotitis. Relapsing symptoms and the presence of parotitis seemed to be characteristic of tumors of Stensen's duct. For a mass accompanied by obstructive parotitis or relapsing parotitis without salivary calculus, a tumor of Stensen's duct should be included in the differential diagnosis. PMID- 17111275 TI - Leiomyoma of the trachea: CT and MRI findings. AB - A 47-year-old woman with a tracheal leiomyoma is described. Leiomyoma is a rare benign tumor of the trachea. Only 43 cases have been reported in the English- and Japanese-language literature. To our knowledge, this is the second time magnetic resonance imaging (MRI) findings have been reported. MRI can help discriminate between masses with a stalk and masses with a broad base, which in turn can help determine whether surgical resection of the tracheal leiomyoma is appropriate. PMID- 17111276 TI - Quality of life after surgery for genital prolapse in elderly women: obliterative and reconstructive surgery. AB - The objective of this study was to determine if obliterative and reconstructive vaginal surgery for advanced pelvic organ prolapse improve quality of life in elderly women. Women age 65 years or older with stage 3 or 4 pelvic organ prolapse who desired surgical correction were prospectively enrolled. The subjects underwent either obliterative or reconstructive vaginal surgery based on their personal preference and sexual expectations. The subjects received a pelvic organ prolapse quantitation examination and completed the pelvic floor distress inventory (PFDI), the pelvic floor impact questionnaire (PFIQ), the SF-36, and the Beck depression inventory preoperatively, 6 and 12 months after surgery. Seventy-nine subjects were enrolled, 70 of whom completed follow-up: 30 in the obliterative group and 40 in the reconstructive group. Both groups demonstrated significant improvements in the pelvic organ prolapse, urinary, and colorectal scales of the PFDI and PFIQ 6 and 12 months after surgery with no differences between the two treatment groups. In addition, there were significant and clinically important improvements noted in the bodily pain, vitality, social functioning, role-emotional, and mental health summary scales of the SF-36 in both groups after surgery, with no significant difference between groups. In appropriately selected elderly women, both obliterative and reconstructive vaginal surgery for advanced pelvic organ prolapse significantly improved health related quality of life. PMID- 17111277 TI - An effective and low-cost culture medium for isolation and growth of Xylella fastidiosa from citrus and coffee plants. AB - Buffered charcoal-yeast extract medium (BCYE) has been used for isolation of Xylella fastidiosa from citrus (Citrus sinensis) and coffee (Coffea arabica) plants affected by citrus variegated chlorosis (CVC) and coffee leaf scorch (CLS). BCYE is composed of ACES (2-[2-amino-2oxoethyl) amino]-ethanesulfonic acid) buffer, activated charcoal, yeast extract, L-cysteine, ferric pyrophosphate, and agar. ACES buffer is costly and not always commercially available in Brazil, and the L-cysteine and ferric pyrophosphate need to be filter sterilized in 0.22-mum pore membranes before inclusion in the medium. Omission of L-cysteine, addition of magnesium sulfate, and replacements of ACES and ferric pyrophosphate for potassium phosphate and ferrous sulfate resulted in an effective, less expensive, and entirely autoclavable medium, named phosphate buffered charcoal-yeast extract medium (PCYE). The final cost of PCYE was approximately one tenth that of BCYE. Its effectiveness was tested for the isolation of X. fastidiosa from symptomatic leaves collected from 52 citrus plants affected by CVC and 43 coffee plants affected by CLS. PCYE was as effective as BCYE and has been used routinely in our and other laboratories for isolation, growth, and quantification of X. fastidiosa from plant tissues. PMID- 17111278 TI - Surgeon responsibility in the era of "outrageous science". PMID- 17111279 TI - Routine laparoscopic cholecystectomy after endoscopic sphincterotomy for choledocholithiasis in octogenarians: is it worth the risk? AB - BACKGROUND: No unanimous consensus has been reached as to the need for routine laparoscopic cholecystectomy (LC) after endoscopic sphincterotomy (ES) for choledocholithiasis in very elderly patients, who are considered as high-risk subjects for surgery. METHODS: From 1991 through 1997, 170 patients were referred to undergo preoperative ES and routine LC for common bile duct (CBD) stones. The results for 27 patients (age 80 years or older) were compared with those achieved for younger patients. Successively, in a retrospective case-control study, the results for the selected patients were compared with those for 27 very elderly patients who underwent endoscopic retrograde cholangiopancreatography (ERCP), but did not receive LC. The mean follow-up period was 126 months. RESULTS: Octogenarians showed longer surgery time (79 vs 51 min) and postoperative hospital stay (2.8 vs 1.2 days), as well as more early low-grade complications (15% vs 3%), whereas there were no differences in conversion rate or serious complications. Recurrent symptoms or complications developed in 48% of octogenarians not undergoing routine LC, and 30% finally needed surgery. One patient in the control group died after emergency cholecystectomy for acute cholecystitis. The results of surgery were significantly poorer for the control group. CONCLUSIONS: Although a "wait-and-see" policy allowed two-thirds of LCs to be avoided in octogenarians, biliary-related events developed for every second patient, often requiring delayed surgery, with poorer results. Sequential treatment (ES followed by elective LC) is a safe procedure for octogenarians, and should be considered as a standard, definitive treatment for cholecystocholedocholithiasis even after the age of 80 years. PMID- 17111280 TI - Attempted establishment of proficiency levels for laparoscopic performance on a national scale using simulation: the results from the 2004 SAGES Minimally Invasive Surgical Trainer-Virtual Reality (MIST-VR) learning center study. AB - BACKGROUND: The Minimally Invasive Surgical Trainer-Virtual Reality (MIST-VR) has been well validated as a training device for laparoscopic skills. It has been demonstrated that training to a level of proficiency on the simulator significantly improves operating room performance of laparoscopic cholecystectomy. The purpose of this project was to obtain a national standard of proficiency using the MIST-VR based on the performance of experienced laparoscopic surgeons. METHODS: Surgeons attending the Society of American Gastrointestinal Endoscopic Surgeons (SAGES) 2004 Annual Scientific Meeting who had performed more than 100 laparoscopic procedures volunteered to participate. All the subjects completed a demographic questionnaire assessing laparoscopic and MIST-VR experience in the learning center of the SAGES 2004 meeting. Each subject performed two consecutive trials of the MIST-VR Core Skills 1 program at the medium setting. Each trial involved six basic tasks of increasing difficulty: acquire place (AP), transfer place (TP), traversal (TV), withdrawal insert (WI), diathermy task (DT), and manipulate diathermy (MD). Trial 1 was considered a "warm-up," and trial 2 functioned as the test trial proper. Subject performance was scored for time, errors, and economy of instrument movement for each task, and a cumulative total score was calculated. RESULTS: Trial 2 data are expressed as mean time in seconds in Table 2. CONCLUSION: Proficiency levels for laparoscopic skills have now been established on a national scale by experienced laparoscopic surgeons using the MIST-VR simulator. Residency programs, training centers, and practicing surgeons can now use these data as guidelines for performance criterion during MIST-VR skills training. PMID- 17111281 TI - Transanal endoscopic microsurgery for carcinoma of the rectum. AB - BACKGROUND: The authors present their experience with rectal cancers managed by transanal endoscopic microsurgery (TEM). METHODS: This prospective study investigated patients undergoing primary TEM excision for definitive treatment of rectal cancer between January 1996 and December 2003 by a single surgeon in a tertiary referral colorectal surgical unit. RESULTS: For this study, 52 patients (30 men and 22 women) underwent TEM excision of a rectal cancer. Their mean age was 74.3 years (range, 48-93 years). The median diameter of the lesions was 3.44 cm (range, 1.6-8.5 cm). The median distance of the lesions from the anal verge was 8.8 cm (range, 3-15 cm), with the tumor more than 10 cm from the anal verge in 36 patients. The median operating time was 90 min (range, 20-150 min), and the median postoperative stay was 2 days. All patients underwent full-thickness excisions. There were 11 minor complications, 2 major complications, and no deaths. The mean follow-up period was 40 months (range, 22-82 months). None of the pT1 rectal cancers received adjuvant therapy. Eight patients with pT2 rectal cancer and two patients with pT3 rectal cancer received postoperative adjuvant therapy. The overall local rate of recurrence was 14%, and involved cases of T2 and T3 lesions, with no recurrence after excision of T1 cancers. Three patients died during the follow-up period, but no cancer-specific deaths occurred. CONCLUSIONS: The findings warrant the conclusion that TEM is a safe, effective treatment for selected cases of rectal cancer, with low morbidity and no mortality. The TEM procedure broadens the range of lesions suitable for local resection to include early cancers (pTis and pT1) and more advanced cancers only in frail people. PMID- 17111282 TI - Technical feasibility of laparoscopic Finney pyloroplasty examined in a canine model. AB - The efficacy of laparoscopic surgery in the treatment of chronic duodenal ulcer has been demonstrated using minimally invasive approaches. This study aimed to evaluate the technical feasibility of laparoscopic Finney pyloroplasty in six dogs. Under laparoscopic guidance, the anastomosis was created using mechanical devices (n = 3) and combining a linear stapler device and intracorporeal suturing (n = 3). The operative time and complications were recorded. Evaluation of the anastomosis included studies of intraoperative and postoperative endoscopy and ultrasonography as well as the complete gastric emptying time. The animals were killed 4 weeks after surgery. Measurement of luminal diameter, amount of adhesion formation, degree of healing, and inflammation or fibrosis were evaluated in the postmortem studies. The operation was successfully completed for all the animals. No intraoperative or postoperative complications were observed. Gastric emptying was significantly enhanced in the postoperative period, as compared with the preoperative results. After 1 month, there was no evidence of anastomotic leak dehiscence at the gastroduodenal anastomosis. Luminal diameter was increased, and no abnormal findings were encountered during the postmortem abdominal exploration. The technical feasibility of performing a safe laparoscopic pyloroplasty in a dog model was demonstrated in this study. PMID- 17111283 TI - Day case laparoscopic incisional hernia repair is feasible, acceptable, and cost effective. AB - BACKGROUND: Day case surgery is increasingly performed in the United Kingdom. Laparoscopic techniques have increased the number of conditions suitable for a day surgical approach. Findings have shown that laparoscopic incisional hernia repair (LIHR) is superior to conventional open techniques. This study aimed to show that day case LIHR is safe, produces a good clinical outcome, and is cost effective. METHODS: Day case laparoscopic repair was performed for 31 consecutive patients (10 men; median age, 67 years; range, 39-80 years). Data were entered prospectively into a database. Patients were discharged within 8 h committed to a 10-day course of oral diclofenac 50 mg three times daily and 2 tablets of codydramol four times daily. Follow-up evaluation was by telephone consultation. Hospital costs for LIHR and open repair were compared. RESULTS: All procedures were completed laparoscopically on a day case basis. Additional unsuspected defects were found in eight cases (25.8%). The median mesh size was 140 cm2 (range, 25-375 cm2), and the median body mass index (BMI) was 28.7 kg/m2 (range, 20-37.1 kg/m2). Operations were performed or supervised by a single consultant surgeon (S.J.W.). Six postoperative seromas resolved spontaneously. Two port-site infections required oral antibiotics, and one diathermy pad burn healed with simple dressings. The median analgesia requirement was 7 days (range, 0-152 days). There were no recurrences during a median follow-up period of 15 months (range, 3-24 months). There was a saving of 616 pounds sterling per procedure. CONCLUSIONS: Day case laparoscopic repair of incisional hernias is feasible and safe and has a good clinical outcome. The hospital costs are less than for open techniques. PMID- 17111284 TI - Thirteen years' experience with laparoscopic transcystic common bile duct exploration for stones. Effectiveness and long-term results. AB - BACKGROUND: The aim of the present study was to evaluate the effectiveness and long-term results of laparoscopic transcystic common bile duct exploration (TC CBDE). METHODS: Ductal stones were present in 344 of 3212 patients (10.7%) who underwent laparoscopic cholecystectomy (LC). The procedure was completed laparoscopically in 329 patients (95.6%), with TC-CBDE performed in 191 patients (58.1%) who are the object of this study, or with a transverse choledochotomy in 138 cases (41.9%). RESULTS: Biliary drainage was employed in 71 of 191 cases (37.2%). Major complications occurred in 10 patients (5.1%), including retained stones in 6 (3.1%). Mortality was nil. No patients were lost to follow-up (median: 118.0 months; range: 17.6-168 months). No signs of bile stasis, no recurrent ductal stones and no biliary stricture were observed. At present 182 patients are alive with no biliary symptoms; 9 have died from unrelated causes. CONCLUSIONS: Long-term follow-up after laparoscopic TC-CBDE proved its effectiveness and safety for single-stage management of gallstones and common bile duct stones. PMID- 17111285 TI - Laparoscopic treatment of Barrett's esophagus: long-term results. AB - BACKGROUND: Gastroesophageal reflux disease (GERD) is considered the main etiologic process in the metaplastic development of Barrett's esophagus (BE). The most serious complication of BE is the possible dysplastic evolution to esophageal carcinoma. Many treatments have been described to prevent the progression of BE. The outcomes of these interventions are controversial. The aim of this study was to assess whether laparoscopic fundoplication for GERD had an impact on the development of BE. METHODS: Prospective data were collected from patients who were treated with a laparoscopic fundoplication for BE. Data was collected and analyzed for a variety of clinical and pathologic outcomes. RESULTS: Laparoscopic fundoplications were completed between 1993 and 2001, with a total sample size of 92 (mean age 53 +/- 11.8 years). Each patient was diagnosed with GERD associated with BE confirmed by both endoscopy and biopsy. A laparoscopic fundoplication was performed in all patients (360 degree fundoplication in 81 patients and partial fundoplication in 11 patients). There was no postoperative mortality or major complications from the procedure. The mean postoperative stay was 3 +/- 1 days. Seventy patients (76% of the overall sample size) were followed up for a mean 4.2 +/- 2.6 years. Of the patients available for follow-up, 33% (n = 23) had a complete regression of their BE; 21% (n = 15) had a decrease in the degree of metaplasia/dysplasia; 39% (n = 27) had no significant change; and 7% (n = 5) experienced a progression of the BE. Five patients required further procedures for three reasons: (1) GERD recurrence (n = 2), (2) progression of BE (n = 2), and (3) intrathoracic migration (n = 1). No patients developed high-grade dysplasia or esophageal carcinoma. CONCLUSIONS: The results of this study suggest that laparoscopic fundoplication offers a safe and effective long-term treatment for BE. The procedure also demonstrated regression of BE in more than 50% of the sample size. PMID- 17111286 TI - Right aortic arch, isolated left subclavian artery and ductus arteriosus with normal intracardiac anatomy: rare manifestation of chromosome 22q11 deletion. PMID- 17111287 TI - Fatal cephalosporin-induced acute hypersensitivity myocarditis. AB - A frequently fatal, although rare, side effect of cephalosporin antibiotics is noninfectious myocarditis. We report two cases of hypersensitivity myocarditis secondary to administration of cephalosporin antibiotics. In both cases, acute hypersensitivity myocarditis was not suspected clinically, and the diagnosis was made postmortem. Histology revealed intense eosinophilic infiltration of the endomyocardium with eosinophil degranulation and myocyte damage, Clinically, death in both cases was due to cardiac failure. When suspected early, appropriate management may be lifesaving. PMID- 17111288 TI - Aortic stenosis: the spectrum of practice. AB - There is significant variation in practice patterns in managing congenital aortic valve stenosis. Review of medical literature reveals no significant information regarding the current practice methods in the treatment of a simple lesion such as aortic stenosis (AS). Therefore, this survey-based study was conducted in an attempt to better understand the uniformity or heterogeneity of practice in treating AS. A questionnaire was prepared to evaluate the style of management of AS. This survey was designed to assess the practice of follow-up visitations, type and frequency of investigative studies, pharmacological therapy, and exercise recommendations. Questions about therapeutic intervention included those of timing and type of intervention. Questionnaires were sent to all academic pediatric cardiology programs in the United States (48 program) and selected international programs from Europe, Asia, and Australasia (19 program). The total number of surveys sent out was 67, and the total number of respondents was 25 (37%), 15 (31%) from the United States and 9 (53%) from outside the United States. The definition of moderate AS varied among respondents. The range provided for mild AS was identified as that with a peak-to-peak pressure gradient of < 25-30 mmHg, peak instantaneous Doppler gradient of < 36-50 mmHg, or mean Doppler gradient of < 25-40 mmHg. On the other hand, severe AS was defined as that with a peak-to-peak gradient of > 50-60 mmHg, peak instantaneous Doppler gradient of > 64-80 mmHg, or mean Doppler gradient of > 45-64 mmHg. In assessing follow-up patterns, 84% of respondents recommended seeing patients with mild AS annually, the longest time of follow-up listed in the questionnaire, whereas 20% suggested follow-up every 6 months. There was no consensus among survey centers regarding follow-up of patients with moderate AS. For severe AS, 16% recommend immediate intervention, 16% arrange follow-up every 6 months, and 56 and 28% recommend follow-up in 3 and 1 month(s), respectively. In making the decision to proceed with biventricular versus univentricular repair in patients with AS in the neonatal period, many factors were considered. Ninety-two percent of respondents rely on mitral valve z score, 84% on aortic valve z score, 52% on left ventricle length, 48% on the presence of antegrade ascending aorta flow, and only 32% considered significant endocardial fibroelastosis as a factor. Rhodes score was used by 20% of respondents in decision making regarding the approach to management of this subset of AS. This study shows that there is consensus in the management of mild and severe forms of AS. As expected, disagreement is present in the definition, evaluation, and therapy of moderate aortic valve stenosis. There is a tendency for catheter intervention except in the presence of dysplastic aortic valve or moderate to severe aortic regurgitation. There is also disagreement regarding methods used to determine biventricular versus univentricular repair of a borderline hypoplastic left heart. PMID- 17111289 TI - Is family history of premature cardiovascular diseases appropriate for detection of dyslipidemic children in population-based preventive medicine programs? CASPIAN study. AB - The objectives of this study were to determine the prevalence of dyslipidemia and the usefulness of self-report family history (FH) of premature cardiovascular disease (CVD) for identifying children with lipid disorders. This study was conducted on a representative, population-based sample of 4811 Iranian children and adolescents (2248 boys and 2563 girls) aged 6-18 years. We compared the obtained serum lipid profile with that of the Lipid Research Clinic (LRC) and calculated the predictive value of FH for detecting those children with dyslipidemia.Overall, for both genders and for age groups, the mean serum triglycerides (TG) and its percentiles were significantly higher, and the mean and percentiles of total, low-density, and high-density cholesterol (TC, LDL-C, and HDL-C respectively) were significantly lower than the LRC values. In total, 45.7% of participants had dyslipidemia; the most frequent ones were low HDL-C (24.8%) and hypertriglyceridemia (24.5%), followed by hypercholesterolemia (6.4%) and high LDL-C (6.3%), respectively. The mean serum lipid levels and the anthropometric measures were not significantly different among those with or without positive FH. The sensitivity, and specificity, positive and negative predictive values for FH in detecting those children with dyslipidemia were 28.4, 70.3. 44.7, and 53.8%, respectively. The usefulness of FH in identifying dyslipidemic children was relatively low. The common lipid disorders in our community were the components of the metabolic syndrome. We suggest that the current guidelines for screening lipid disorders in youths, which are based on cholesterol, should consider such ethnic differences. PMID- 17111290 TI - Bartonella henselae endocarditis in a child. AB - 14-year-old white male with a past medical history of congenital bicuspid aortic valve, Streptococcus viridans endocarditis, and pulmonary valve homograft presented with culture-negative endocarditis. Molecular studies identified the causative organism as Bartonella henselae and subsequent serologic studies supported this diagnosis. This rare cause of endocarditis may be under recognized in children. Bartonella henselae endocarditis should be considered in all children with culture-negative endocarditis, and molecular studies using a polymerase chain reaction-based assay should be routinely utilized in the evaluation of all endocarditis cases that are culture negative to rapidly diagnosis this treatable cause of endocarditis. PMID- 17111291 TI - Efficacy of prostaglandin E1 in relieving obstruction in coarctation of a persistent fifth aortic arch without opening the ductus arteriosus. AB - A persistent fifth aortic arch (PFAA) with coarctation and type A interruption of the fourth arch was recognized in a 9-day-old infant. The widening of the area of coarctation in the fifth arch with prostaglandin E1 infusion without opening the ductus arteriosus is presented to document that ectopic ductal tissue constriction contributes to the development of coarctation in PFAA. PMID- 17111292 TI - Use of Amplatzer fenestrated atrial septal defect device in a child with familial pulmonary hypertension. AB - In a 4.5-year-old child with refractory pulmonary arterial hypertension, we performed atrial septostomy with the application of an Amplatzer fenestrated device designed to maintain patency. Continuous intravenous epoprostenol infusion was started concomitantly. Forty-two months after the procedure, the patient had no recurrent syncope and remained in New York Heart Association functional class II. Fenestration of the atrial septum is feasible in children with pulmonary artery hypertension. No conclusion regarding the patient's need for an atrial septal defect can be drawn since concomitant prostanoid therapy was administered. The long-term patency of the atrial communication needs further confirmation and the optimal timing for its application has to be determined. PMID- 17111293 TI - Echocardiographic follow-up of congenital aortic valvular stenosis. AB - We investigated the morphology of the stenotic aortic valve, the progression of the stenosis, and the onset and progression of aortic regurgitation (AR) in patients with congenital aortic valvular stenosis (AVS). The medical records of 278 patients with AVS were reviewed, with the patients with concomitant lesions besides AR excluded. Very mild aortic stenosis was defined as a transvalvular Doppler peak systolic instantaneous gradient (PSIG) less than 25 mmHg, mild stenosis as 25-49 mmHg, moderate stenosis as 50-75 mmHg, and severe stenosis as more than 75 mmHg. The mean age of the patients was 4.9 +/- 4.3 years (range, 3 days to 15 years), and 203 (73%) were male. The number of the cusps was determined with two-dimensional echocardiography in 266 patients (95%): unicuspid in 3 patients (1%), bicuspid in 127 patients (48%), and tricuspid in 136 patients (51%). A total of 192 of all patients were followed for 2 months to 14.6 years (mean 4.2 +/- 3.3 years) with medical treatment alone. Among 72 patients with very mild stenosis at initial echocardiographic examination, 20% had mild, 3% moderate, and 1% severe stenosis after a mean period of 3.7 years. In 70 patients with mild stenosis at initial echocardiographic examination, 28% had moderate and 9% severe stenosis after a mean period of 5 years. Among 44 patients with moderate stenosis at initial echocardiographic examination, 36% had severe stenosis after a mean period of 3.7 years. Among 192 patients, 40% had AR (3% trivial, 28% mild, and 9% moderate) at initial echocardiographic examination. After a mean period of 4.2 years, 58% of the patients had AR (13 % trivial, 25% mild, 16% moderate, and 4% severe). There was not statistically significant difference between catheterization peak systolic gradients (47 +/- 16 mmHg) and Doppler estimated mean gradients (45 +/- 9 mmHg) (p = 0.53), whereas Doppler PSIGs (74.9 +/- 15.7 mmHg) were higher than catheterization peak systolic gradients (p < 0.0001) in 25 patients who were studied in the catheterization lab. Patients with very mild stenosis may be followed with a noninvasive approach every 1 or 2 years, and an annual follow-up is suggested for patients with mild stenosis. Nearly one-third of patients with moderate stenosis at initial echocardiographic examination had severe stenosis after a mean period of 3.7 years. Therefore, we recommend, that patients with moderate stenosis undergo noninvasive evaluation every 6 months. Doppler estimated mean gradient is very useful in predicting the need for intervention in children with AVS. PMID- 17111294 TI - Echocardiographic follow-up of children with isolated discrete subaortic stenosis. AB - This study evaluates the progression of stenosis, onset and progression of aortic regurgitation (AR), and the results of surgical outcomes in children with isolated discrete subaortic stenosis (SAS). The medical records of 108 patients (mean age, 5.5 +/- 3.8 years; range, 3 days to 18 years) with isolated discrete SAS were reviewed. Patients with lesions other than AR were excluded. Very mild stenosis was defined as Doppler peak systolic instantaneous gradient (PSIG) less than 25 mmHg, mild stenosis as 25-49 mmHg, moderate stenosis as 50-75 mmHg, and severe stenosis as more than 75 mmHg. Seventy-eight of 108 patients were followed for 2 months to 14 years (mean, 4.8 +/- 3.7 years; median, 5 years) with medical treatment alone. In these patients, the mean PSIG at last echocardiogram was higher than the mean PSIG at initial echocardiogram (39 +/- 19 vs 31 +/- 12 mmHg, respectively; p < 0.001). Among 24 patients with very mild stenosis at initial echocardiogram, 10 had mild and 2 had moderate stenosis after a mean period of 5.6 years. Among 46 patients with mild stenosis at initial echocardiogram, 11 had moderate and 5 had severe stenosis after a mean period of 4.1 years. Only 1 patient among the 8 patients with moderate stenosis at initial echocardiogram had severe stenosis after a mean period of 2.7 years. Thirty-nine patients (50%) had AR (13% trivial, 33% mild, and 4% moderate) at initial echocardiogram. After a mean period of 4.8 years, 77% of the patients had AR (10% trivial, 53% mild, 9% mild-moderate, and 5% moderate). Twenty-four patients underwent surgery. Preoperatively, mean Doppler PSIG and AR incidence were 64 +/- 17 mmHg and 91% (22/24), respectively. The mean Doppler PSIG was 30 +/- 19 mmHg and AR was present in all of the patients a mean period of 4.1 years after surgery. Two patients underwent reoperation for recurrent SAS and AR. Patients with very mild or mild stenosis may be followed noninvasively every year. One patient of the 8 patients with moderate stenosis progressed to severe stenosis, and moderate AR developed in 2 patients after a mean of 2.7 years. We recommend that patients with moderate stenosis undergo careful evaluation to determine whether surgery is necessary due to the severity of stenosis and AR. PMID- 17111295 TI - Prevalence of arrhythmias and their risk factors mid- and long-term after the arterial switch operation. AB - Early results of the arterial switch operation (ASO) for transposition of the great arteries (TGA) are good, but there are few mid- and long-term data on postoperative arrhythmias, especially in Japan. In this study, clinical data on 624 1-year survivors who had an ASO between 1976 and 1995 were collected from six institutes in Japan up to October 2002. Sixty (9.6%) 1-year survivors had significant arrhythmias. Bradycardia occurred in 22 patients, including complete atrioventricular block (CAVB) in 12, sick sinus syndrome (SSS) in 6, and second degree atrioventricular block in 4. Syncope developed in 2 with CAVB and 2 with SSS. Ten patients with bradycardia underwent permanent pacemaker implantation. Supraveutricular tachycardia (SVT) was seen in 25 patients, including paroxysmal supraventricular tachycardia in 16, atrial flutter in 7, and atrial fibrillation in 2. Six patients with SVT received antiarrhythmic medication. SVT was transient in 20 and persistent in 5. Ventricular arrhythmias occurred in 13 patients, including nonsustained ventricular tachycardia in 5, paroxysmal ventricular contractions with couplets in 5, ventricular flutter in 2, and sustained ventricular tachycardia in 1. Four patients with ventricular arrhythmias received antiarrhythmic medication. Of the study patients, 8 died 1 year or more after ASO. Death was directly related to arrhythmia in 1 patient and was due to nonsustained ventricular tachycardia with severe congestive heart failure. The presence of a ventricular septal defect (VSD) was a risk factor for postoperative arrhythmia. Patients with TGA and VSD had more arrhythmias than those with TGA and an intact ventricular septum (13.7 vs 8.7%, p < 0.05), and this was especially true for CAVB (3.9% vs 1.0%, p < 0.05). In 36 patients clearly documented time onset of postoperative arrhythmia arrhythmia developed in 18 (50%) after less than 1 year and in 15 (42%) after more than 5 years. In summary serious arrhythmias after ASO were uncommon, but postoperative arrhythmias, such as unpaced CAVB, SSS, and VT, were related to morbidity and mortality. VSD was a risk factor for postoperative arrhythmia, especially CAVB. Approximately half of the arrhythmias developed late. Lifelong monitoring with respect to arrhythmia is needed for patients after ASO. PMID- 17111296 TI - Macroangiopathy in adults and children with diabetes: from molecular mechanisms to vascular damage (part 1). AB - Type 2 diabetes mellitus (T2DM) is an increasing problem in childhood; however type 1 diabetes mellitus (T1DM) remains by far the most common type of diabetes in this age group. In this review we will focus on T1DM, because this will have the greatest implication for patients diagnosed in childhood. During the atherosclerotic process, several molecular, receptorial and cellular factors provide a continous mechanism of vascular damage. In diabetic children this state seems to be enhanced and facilitated so that accelerated atherosclerosis is associated with an increased risk of cardiovascular events in respect to the non diabetic population. Hyperglycemia PER SE and associated with diabetes is an important risk factor for atherosclerosis. At present a substantial part of children with diabetes do not reach satisfactory glycemic control. Other risk factors for the development and progression of atherosclerosis may be inherited or develop in the course of the disease: hypertension, dyslipidemia, insulin resistance, obesity, cigarette smoking, physical inactivity, disturbance of platelet function, coagulation and fibrinolysis. The development and progression of atherosclerosis should be blocked at an early age, if possible. Primary prevention to all risk factors for cardiovascular disease is important and intervention is indicated if necessary. At the moment the best therapeutic strategy is to maintain metabolic control at a physiologic level and perform screening and early intervention for vascular complications. PMID- 17111297 TI - Macroangiopathy in adults and children with diabetes: risk factors (part 2). AB - Autoimmune or type 1 diabetes mellitus (T1DM), accounts for 90-95% of all cases of diabetes, while type 2 diabetes mellitus (T2DM), characterized by impaired insulin sensitivity and production, accounts for the other 5-10%. Atherosclerotic process starts during childhood and recognize several mechanisms that are activated in response to NOXIUS STIMULI and participate in a complex state which is accepted to be a chronic inflammatory state. T1DM patients, especially those with a non-optimal metabolic control, have a higher risk of developing all macrovascular complications such as myocardial infarction, stroke and silent ischemia. Macrovascular disease is mainly associated with hyperglycemia, dyslipidemia, obesity, hypertension, hypercoagulable state, cigarette smoking, lack of exercise, endothelial dysfunction, hyperhomocysteinemia and vascular wall abnormalities. In this paper we review the importance of traditional and non traditional risk factors for macrovascular complications in children with T1DM and discuss their role in the pathogenesis of the excess cardiovascular mortality in these patients. PMID- 17111298 TI - Fatty acids inhibit intramyocellular triglyceride synthesis and turnover acutely in high fat-fed obese rats. AB - Obesity is associated with hyperlipidemia and enlarged intramyocellular triglyceride (imcTG) stores. The latter is strongly correlated with muscle insulin resistance. However, whether hyperlipidemia plays a role in imcTG accumulation is unknown. In the present study, the effects of plasma fatty acids on imcTG fractional turnover rate (FTR) and synthesis in skeletal muscle of high fat-fed obese rats have been examined using pulse-chase technique. imcTG was prelabeled (pulse) by continuous infusion of U- (14)Cglycerol and then the loss of (14)C-labels from imcTG was chased while exogenous fatty acids were infused at 0 (saline), 1 (L) or 3 (H) micromol/kg/min. imcTG synthesis was determined using 2- (3)Hglycerol during the chase. L and H fatty acid infusions raised plasma fatty acids by 14% (p=0.02) and 30% (p=0.001), respectively, while plasma insulin and glycerol and the rate of glycerol appearance remained unchanged (p>0.05). imcTG FTR was suppressed by 36-40% and 48% in gastrocnemius and tibialis anterior, respectively (both p<0.05), and imcTG synthesis was suppressed by 50 60% in the same muscles (both p<0.05). In contrast, neither turnover nor synthesis of imcTG in soleus was affected by fatty acid infusion (p>0.05). imcTG content and the activities of diglyceride acyltransferase and hormone sensitive lipase were not affected by fatty acid infusion. The findings suggested that elevated plasma fatty acids suppress imcTG turnover and synthesis simultaneously and thus do not appear to promote imcTG accumulation in this obesity model at least in short term. PMID- 17111299 TI - The primary amine metabolite of sibutramine stimulates lipolysis in adipocytes isolated from lean and obese mice and in isolated human adipocytes. AB - Sibutramine is a satiety-inducing serotonin-noradrenaline reuptake inhibitor that acts predominantly via its primary and secondary metabolites. This study investigates the possibility that sibutramine and/or its metabolites could act directly on white adipose tissue to increase lipolysis. Adipocytes were isolated by a collagenase digestion procedure from homozygous lean (+/+) and obese diabetic OB/OB mice, and from lean nondiabetic human subjects. The lipolytic activity of adipocyte preparations was measured by the determination of glycerol release over a 2-hour incubation period. The primary amine metabolite of sibutramine M2, caused a concentration-dependent stimulation of glycerol release by murine lean and obese adipocytes (maximum increase by 157+/-22 and 245+/-16%, respectively, p<0.05). Neither sibutramine nor its secondary amine metabolite M1 had any effect on lipolytic activity. Preliminary studies indicated that M2 induced lipolysis was mediated via a beta-adrenergic action. The non-selective beta-adrenoceptor antagonist propranolol (10 (-6) M) strongly inhibited M2 stimulated lipolysis in lean and obese murine adipocytes. M2 similarly increased lipolysis by isolated human omental and subcutaneous adipocytes (maximum increase by 194+/-33 and 136+/-4%, respectively, p<0.05) with EC50 values of 12 nM and 3 nM, respectively. These results indicate that the sibutramine metabolite M2 can act directly on murine and human adipose tissue to increase lipolysis via a pathway involving beta-adrenoceptors. PMID- 17111300 TI - Generation of reactive oxygen species by endothelial and smooth muscle cells: influence of hyperglycemia and metformin. AB - There is evidence that reactive oxygen intermediates (ROI) play an important role in the pathogenesis of vascular complications in diabetes. On the other hand, metformin, one of the most often used antidiabetic compounds has not only been shown to reduce the risk for vascular complications, but in addition these protective effects are largely independent of its well-known antihyperglycemic action. Therefore, to explain the vasculoprotective effects of metformin, a direct antioxidative action of this compound has been suggested. We show here that human endothelial cells (HUVEC) generate ROI not only in response to high glucose (30 mmol/l glucose), but also in response to palmitic acid, and advanced glycation end-products (carboxymethyllysine and S100 proteins). Metformin inhibited the production of ROI in response to all these stimuli. By double staining-dichlorofluorescein as marker of ROI and Mitotracker CMH-Ros for mitochondria-the mechanism of ROI generation was analyzed in more detail in smooth muscle cells. Our data suggest that ROI are generated by uncoupling of the mitochondrial respiratory chain as well as by activation of the cytosolic NADPH oxidase. A complete inhibition of ROI generation is only achieved by simultaneous inhibition of the mitochondrial electron flux (theonyltrifluoroacetone) and NADPH oxidase (apocynin). Our data suggest that the various processes contributing to generation of ROI are closely linked. Activation of AMP kinase may represent an important mechanism to understand the antioxidative effects of metformin on the mitochondrial and cytosolic generation of ROI. PMID- 17111301 TI - Smad3 differently affects osteoblast differentiation depending upon its differentiation stage. AB - Smad3, a critical component of the TGF-beta signaling pathways, plays an important role in the regulation of bone formation. However, how Smad3 affects osteoblast at the different differentiation stage remains still unknown. In the present study, we examined the effects of Smad3 on osteoblast phenotype by employing mouse bone marrow ST-2 cells and mouse osteoblastic MC3T3-E1 cells at the different differentiation stage. Smad3 overexpression significantly inhibited bone morphogenetic protein-2 (BMP-2)-induced ALP activity in ST-2 cells, indicating that Smad3 suppresses the commitment of pluripotent mesenchymal cells into osteoblastic cells. Smad3 increased the levels of COLI and ALP mRNA at 7 day cultures in MC3T3-E1 cells, and its effects on COL1 were decreased as the culture periods progress, although its effects on ALP were sustained during 21 day cultures. Smad3 overexpression enhanced the level of Runx2 and OCN mRNA at 14 day and 21 day cultures. Smad3 increased the levels of MGP and NPP-1 mRNA, although the extent of increase in MGP and NPP-1 was reduced and enhanced during the progression of culture period, respectively. Smad3 did not affect the level of ANK mRNA. On the other hand, Smad3 enhanced the level of MEPE mRNA at 14 and 21 day cultures, although Smad3 decreased it at 7 day cultures. In conclusion, Smad3 inhibits the osteoblastic commitment of ST-2 cells, while promotes the early stage of differentiation and maturation of osteoblastic committed MC3T3-E1 cells. Also, Smad3 enhanced the expression of mineralization-related genes at the maturation phase of MC3T3-E1 cells. PMID- 17111302 TI - Endocrine disorders in pediatric - onset Langerhans Cell Histiocytosis. AB - Langerhans Cell Histiocytosis (LCH) is a rare disorder with a great variety of clinical manifestations. The purpose of this retrospective study was to evaluate the pattern and the long-term course of clinical, laboratorial and radiological findings in pediatric-onset LCH. We reviewed 46 children with histological diagnosis of LCH. Ten children (22%) showed endocrine disorders. Central diabetes insipidus (DI) was observed in all ten patients; GH deficiency was confirmed in four and hypogonadism in two children. There were no adrenal, prolactin or thyroid axis abnormalities. Obesity was observed in three patients. Eight patients showed soft tissue infiltration and five bone involvement. The MRI showed a lack of posterior pituitary bright spot in all DI patients; infundibular infiltration (II) associated or not with sellar or supra-sellar mass was observed in 4 patients. We conclude that the investigation of LCH, a multi-systemic disease, should include central nervous system images. The presence of II and/or DI should raise the diagnosis of LCH. Complete endocrine evaluation, allowing an early hormone therapy, is required to obtain a better quality of life in children with LCH. PMID- 17111303 TI - Ghrelin response to acute aerobic exercise in boys at different stages of puberty. AB - The aim of this study was to investigate the changes in serum ghrelin and leptin concentrations during acute aerobic cycle ergometer test in 60 boys at different pubertal stages. Boys were divided according to their pubertal status as group I (Tanner stage 1, n=20), group II (Tanner stages 2 and 3, n=20) and group 3 (Tanner stages 4 and 5, n=20). Maximal oxygen consumption and individual ventilatory threshold of the subjects were measured directly using stepwise increasing loads on cycle ergometer. Second exercise test consisted of a 30 minute constant load exercise on the same ergometer at the level of approximately 95% of the individual ventilatory threshold. Venous blood samples were obtained before, immediately after and after 30 minutes of recovery for the measurement of serum ghrelin, leptin, testosterone and insulin. At baseline, prepubertal children had significantly higher values for serum ghrelin compared to the groups II and III. Acute exercise altered significantly only insulin concentration. In all the groups, the maximal oxygen consumption/kg correlated positively with basal levels of testosterone (r=0.60, p<0.001) and insulin (r=0.34), and negatively to ghrelin (r=-0.35) and leptin (r=-0.32) (p<0.05). We conclude that moderate acute aerobic exercise does not change serum ghrelin or leptin level in boys at different pubertal stages. PMID- 17111304 TI - Plasma aminopeptidase activities in Parkinson's disease. AB - Parkinson's disease (PD) is an age-related neurodegenerative disease characterized by a progressive motor disorder, but frequently is accompanied by autonomic symptoms such as hypotension. Together with the decrease of dopamine, significant decreases in aminopeptidase activities have been reported in PD brains. However, up to date there are no studies about changes of aminopeptidase activities in plasma of PD patients. We studied plasma activities of alanyl-, aspartyl-(AspAP), cystinyl-(CysAP) and glutamyl-aminopeptidase (GluAP) in two groups of subjects: control (n=41) and PD (n=48). Plasma activities of AspAP, CysAP, and GluAP showed significant decreases of 24.9% (p<0.05), 39.4% (p<0.01) and 33.3% (p<0.01), respectively, in PD group. These aminopeptidases are involved in the metabolism of circulating peptides such as the ones of the renin angiotensin system. The importance of aminopeptidases in striatal dopamine content and in neuroendocrine system in PD is discussed. PMID- 17111305 TI - Arabinoxylan fibre consumption improved glucose metabolism, but did not affect serum adipokines in subjects with impaired glucose tolerance. AB - The consumption of arabinoxylan, a soluble fibre fraction, has been shown to improve glycemic control in type 2 diabetic subjects. Soluble dietary fibre may modulate gastrointestinal or adipose tissue hormones regulating food intake. The present study investigated the effects of arabinoxylan consumption on serum glucose, insulin, lipids, leptin, adiponectin and resistin in subjects with impaired glucose tolerance. In a randomized, single-blind, controlled, crossover intervention trial, 11 adults consumed white bread rolls as either placebo or supplemented with 15 g arabinoxylan for 6 weeks with a 6-week washout period. Fasting serum glucose, insulin, triglycerides, unesterified fatty acids, apolipoprotein A1 and B, adiponectin, resistin and leptin were assessed before and after intervention. Fasting serum glucose, serum triglycerides and apolipoprotein A-1 were significantly lower during arabinoxylan consumption compared to placebo (p=0.029, p=0.047; p=0.029, respectively). No effects of arabinoxylan were observed for insulin, adiponectin, leptin and resistin as well as for apolipoprotein B, and unesterified fatty acids. In conclusion, the consumption of AX in subjects with impaired glucose tolerance improved fasting serum glucose, and triglycerides. However, this beneficial effect was not accompanied by changes in fasting adipokine concentrations. PMID- 17111306 TI - Increased prolactin in acute coronary syndromes as putative Co-activator of ADP stimulated P-selectin expression. AB - Prolactin and leptin are newly recognized platelet co-stimulators due to enhancement of ADP-induced platelet aggregation. The aim of our study was to assess whether both hormones prolactin and leptin play a role as co-activators of platelet activation in patients with acute coronary syndromes. Twenty-one patients with acute coronary syndromes, 10 with stable angina pectoris and 10 controls were studied. Patients with acute coronary syndromes showed significantly higher prolactin and leptin values and a significant increased P selectin expression on platelets compared to patients with stable angina pectoris or controls. However, patients with acute myocardial infarction as a subgroup of acute coronary syndromes showed the highest prolactin levels as well as ADP stimulated P-selectin expression. In the myocardial infarction subgroup prolactin values showed a significant correlation to ADP stimulated P-selectin expression on platelets (r (2)=0.41; p=0.025), whereas leptin was not correlated. Our data indicate an association between increased prolactin values and enhanced P selectin expression on platelets in patients with acute coronary syndromes. Therefore, the stress hormone prolactin could be a co-stimulator of platelet activation in these patients. In contrast, the putative platelet activator leptin does not seem to play a major role in acute coronary syndromes. PMID- 17111307 TI - Association study of the 804C/A and 252A/G polymorphisms in the lymphotoxin-alpha gene for microalbuminuria in Japanese type 2 diabetic patients. PMID- 17111308 TI - Lower body negative pressure treadmill exercise is more comfortable and produces similar physiological responses as weighted vest exercise. AB - Lower body negative pressure (LBNP) treadmill exercise can generate a hypergravity load on the lower body that may improve athlete performance by mechanical and cardiovascular adaptations. This study compared the cardiovascular responses, subjective exertion and discomfort levels produced by LBNP exercise with those generated by a weighted vest (WV). We hypothesized that LBNP exercise is more comfortable than WV exercise at comparable levels of exercise. Nine subjects exercised on a treadmill at nine conditions, at 5.5 mph for 15 minutes, in which they ran in random order to avoid confounding effects: 100 %, 110 %, 120 %, 130 %, and 140 % body weight (BW), the latter four conditions were achieved by either LBNP chamber or WV. Heart rate (HR) and oxygen consumption (.VO(2)) were monitored continuously using ECG and open circuit spirometry. At the end of each test, subjects were asked to give discomfort and exertion scores using a ten point visual analog scale (10 = maximal discomfort and exertion). For both HR and .VO(2), no significant differences were observed between LBNP and WV. Subjects reported significantly higher discomfort levels when exercising with the WV than with the LBNP at 120 % BW (5.1 +/- 0.55 vs. 3.1 +/- 0.64; p < 0.05), 130 % BW (6.2 +/- 0.42 vs. 2.3 +/- 0.44; p < 0.01) and 140 % BW (6.9 +/- 0.27 vs. 4.7 +/- 0.60; p < 0.01), while maintaining similar exertions at all conditions. Based on these results, LBNP exercise is more comfortable than standard WV exercise, while maintaining similar exertion, HR and .VO(2) values. PMID- 17111309 TI - Effect of pool length on blood lactate, heart rate, and velocity in swimming. AB - Exercise testing in water has been used to follow the progression of conditioning during regular training in swimmers. The present study examined the effects of pool length in eleven male swimmers on a set of 5 x 200-m freestyle swims with increasing speed from submaximal to maximal. Mean velocity of swimming, blood lactate and heart rate were examined in both 25-m and 50-m pools. Turning benefit as a marker for turning skill was measured separately by a underwater video system (speed difference between pre- and post-turning) during short all-out swims. Maximum force during swimming was measured in tethered swimming and explosive strength of leg extensor muscles was evaluated by a counter movement jump. The significantly higher (p=0.033 - 0.000) blood lactate values for the 50 m pool as compared to the 25-m pool were found at each point of swimming velocity versus blood lactate curve. The highest post-test lactate level was 7.36 +/- 1.47 mmol x l (-1) in the short course and 8.24 +/- 1.55 mmol x l (-1) (p=0.033) in the long course. The maximum swimming velocity was significantly greater (4.5 %) in the 25-m pool swimming (1.38 +/- 0.11 m x s (-1) vs. 1.32 +/- 0.12 m x s (-1); p=0.000). The heart rate values were significantly (p=0.020 - 0.000) lower in the short course than in the long course at all points of submaximal velocity with a mean difference of 7.3 +/- 0.7 bpm. Heart rate was equal (172 +/- 14 vs. 172 +/- 14 bpm) after the maximum swims in both short and long course. The turning benefit in the short maximum swim was 0.12 +/- 0.05 m x s (-1) (8.1 +/- 3.2 %), correlating positively with the difference in maximal swimming velocity between the short and long-pool swims (r = 0.59; p = 0.029), with the maximum force during tethered swimming (r=0.75; p=0.004) and with the vertical jumping height in the counter movement jump (r=0.55; p=0.039). We conclude that the pool length has a strong effect on blood lactate concentration and heart rate with greater swimming velocity in the short course pool. PMID- 17111310 TI - Assessment of maximal aerobic power and critical power in a single 90-s isokinetic all-out cycling test. AB - The purpose of this study was to establish the validity of a 90-s all-out test for the estimation of maximal oxygen uptake (V.O (2max)) and submaximal aerobic ability as represented by critical power. We hypothesized that the fall in power output by the end of the 90-s all-out test (end power) would represent the exhaustion of anaerobic work capability, and as such, would correspond with the critical power. Sixteen active individuals (mean +/- SD: 30 +/- 6 years; 69.6 +/- 9.9 kg) carried out a series of tests: (i) an incremental ramp test to determine V.O (2max), (ii) three fixed-work rate trials to exhaustion to determine critical power, and (iii) two 90-s all-out tests to measure end power and peak V.O (2). End power (292 +/- 65 W) was related to (r=0.89) but was significantly higher (p<0.01) than critical power (264 +/- 50 W). The mean +/- 95 % limits of agreement (29 +/- 65 W) were too low to use these variables interchangeably. The peak V.O (2) in the 90-s trial was significantly lower than the V.O (2max) (3435 +/- 682 ml x min (-1) vs. 3929 +/- 784 ml x min (-1); p<0.01); mean +/- 95 % limits of agreement was equal to 495 +/- 440 mL x min (-1). The 90-s all-out test cannot, therefore, assess both V.O (2max) and critical power in adult performers. The duration of all-out exercise required to allow V.O (2) to attain its maximum is longer than 90 s. PMID- 17111311 TI - Lactate threshold does not influence metabolic responses during exercise in cyclists. AB - The purpose of this study was to compare plasma markers of metabolic stress and other physiological parameters during prolonged endurance exercise of different intensity in trained subjects possessing a "high" or "low" lactate threshold (LT) expressed as a percentage (%) of peak power output (PPO). Fifteen trained male cyclists completed an incremental exercise test for determination of PPO and the LT (% PPO). Each subject then completed a 90-min and 20-min exercise trial at an intensity representing 75 and 85 % of PPO, respectively. Blood lactate (La), as well as plasma hypoxanthine (Hx) and uric acid (UA) were measured during each exercise trial. The responses in two groups, one (n = 8) with a LT approximately 65 % PPO (LT (low)), the other group (n = 7) with a LT approximately 75 % (LT (high)) (p < 0.01), were then compared. With the exception of UA, La and Hx increased significantly (p < 0.01) throughout each exercise trial compared to rest. However, there were no significant differences in each trial between the two groups of cyclists. There were also no significant differences in the other physiological parameters in each exercise trial between the subjects in LT (low) and LT (high). This study demonstrates that in trained cyclists homogeneous in terms of PPO, plasma markers of metabolic demand during prolonged exercise are not influenced by the LT when measured in an incremental exercise test. PMID- 17111312 TI - Exercise has a positive effect on endothelial progenitor cells, which could be necessary for vascular adaptation processes. AB - Since many years, it was believed that the sole mechanism for postnatal growth of new blood vessels in response to exercise is angiogenesis, occurring through vascular sprouting and intucesseption. Today, accumulating evidence indicates that peripheral blood contains marrow-derived endothelial progenitor cells, which have the potential to differentiate into mature endothelial cells and which can contribute to postnatal vessel growth and repair (postnatal vasculogenesis). However, controversy exists with respect to the identification and the origin of endothelial progenitor cells, this review focus on the different possible sources of endothelial progenitor cells like hematopoietic stem cells, monocytes/macrophages, and mesenchymal stem cells (multipotent adult progenitor cells; MAPCs). In the last years, several groups were able to show the positive effects of exercise on endothelial progenitor cells, which means that vasculogenesis/endothelial progenitor cells might as well contribute to the growth of new blood vessels in adaptation processes. The effect of exercise on endothelial progenitor cells and possible mobilization-factors like hypoxia are discussed. PMID- 17111313 TI - Estimation of the lactate threshold from heart rate response to submaximal exercise: the pulse deficit. AB - In this study, we evaluated the validity of a sharp increase in pulse deficit (PD) as a noninvasive index for estimation of the first lactate threshold (LT (1)) in healthy individuals with various levels of aerobic fitness. Three groups of healthy male subjects participated in the study: 15 sedentary individuals, 14 students of physical education, and 13 competitive athletes. Each subject performed a maximal incremental exercise test on the cycle ergometer for the determination of the LT (1), the second lactate threshold, and peak power output. On different days, subjects performed several 8-min bouts of constant-load exercise on the cycle ergometer, corresponding to each of the power outputs of the maximal test, to evaluate PD, which was calculated as the total number of heart beats of the last 4 min minus the total number of heart beats in the first 4 min of exercise. The three groups presented similar blood lactate, heart rate and pulse deficit responses to exercise. For the first power output up to the LT (1), PD showed no significant changes. For the three groups, a sharp increase in PD was seen at the intensity immediately above LT (1). There was a significant correlation between PD and blood lactate changes from the rest to 4th min of submaximal exercise (r = 0.83, p < 0.05). The power output before a sharp increase in PD detected during constant-load exercise (112 +/- 38 W) and the power output corresponding to the LT (1) detected during the incremental test (111 +/- 37 W, p = 0.323) were similar and strongly correlated (r = 0.99, p = 0.0001). The absolute cut-point value of 25 beats for PD had a sensitivity of 100 %, a specificity of 95 %, and a positive predictive value of 90 % for the detection of LT (1). The determination of PD provides an accurate noninvasive estimate of the LT (1) in healthy young men with different levels of fitness. One 8-min submaximal exercise bout can establish if an individual is exercising above or below the LT (1). PMID- 17111314 TI - Comparative effects of a high-intensity interval warm-up and salbutamol on the bronchoconstrictor response to exercise in asthmatic athletes. AB - Approximately half of all asthmatics become refractory to exercise-induced bronchoconstriction (EIB) with repeated challenges. Exercise refractoriness has been utilized by asthmatic athletes to reduce the bronchoconstrictor response to exercise prior to competition, and this has led to the observation that some asthmatic athletes can "run through" their asthma. The main aim of this study was to investigate the efficacy of short high-intensity, repeated warm-ups compared with salbutamol (a commonly used inhaled beta (2)-agonist) on the severity of EIB. Eight moderately trained (.VO(2peak), 51.9 +/- 2.3 ml . kg (-1) . min (-1)) recreational asthmatic athletes with documented EIB were tested under 4 experimental conditions: 1) control (CON) condition; 2) an interval warm-up (WU) consisting of 8 x 30-sec runs at peak treadmill speed, with 45-sec recovery between each sprint; 3) inhaling 200 microg of salbutamol (Ventolin, GlaxoSmithKline, Uxbridge, Middlesex, U.K.) (IH); and 4) combining both the WU and IH session. All 4 experimental sessions were followed by an exercise challenge test (85-90 % predicted maximum heart rate for 8 min). Pulmonary function was measured pre-exercise and at 1, 5, 10, 15 min postexercise. The mean maximum percent fall in pre- to postexercise forced expiratory volume in 1-sec (FEV (1)) for all 8 asthmatic subjects during the EIB screening test (CON session) was - 18.25 +/- 4.01 %. The mean maximum percent decrease in postexercise FEV (1) significantly decreased (p < 0.05) to only - 9.1 +/- 0.6 % following the WU condition, which is below the EIB diagnostic threshold of a 10 % fall in postexercise FEV (1). The IH and WU + IH condition resulted in a substantial postexercise bronchodilation as shown by a significant increase (p < 0.05) in the mean maximum percent change in postexercise FEV (1) following the IH (+ 8.9 +/- 6.1 %) and WU + IH (+ 15.2 +/- 4.6 %) condition. Similar changes as a result of experimental condition were observed for FEF (25-75 %). These data indicate that repeated high-intensity warm-ups can lessen the bronchoconstrictor response to exercise. In addition, combining the interval warm-up with salbutamol prior to exercise resulted in substantial bronchodilation and conferred a greater protective effect against developing EIB than either intervention alone. PMID- 17111315 TI - Evidence of decrease in peak heart rate in acute hypoxia: effect of exercise induced arterial hypoxemia. AB - This study focuses on the influence of the arterial oxygen saturation level at exhaustion on peak heart rate under acute moderate hypoxia, in endurance-trained subjects. Nineteen competing male cyclists performed exhaustive ramp exercise (cycle ergometer) under normoxia and normobaric hypoxia (15 % O (2)). After the normoxic trial, the subjects were divided into those demonstrating exercise induced arterial hypoxemia during exercise (> 5 % decrease in SaO (2) between rest and the end of exercise, n = 10) and those who did not (n = 9). O (2) uptake, heart rate and arterial O (2) saturation (ear-oximeter) levels were measured. Under hypoxia, peak heart rate decreased for both groups (p < 0.001) and to a greater extent for hypoxemic subjects (p < 0.01). Arterial O (2) saturation under hypoxia was lower for the hypoxemic than for the non-hypoxemic subjects (p < 0.001) and it was correlated to the fall in peak heart rate between normoxia and hypoxia for all subjects (p < 0.01; r = 0.65). Hypoxemic subjects presented greater decrease in maximal O (2) uptake than non-hypoxemic ones (19.6 vs. 15.6 %; p < 0.05). The results confirm the greater decrement in arterial O (2) saturation under hypoxia in hypoxemic subjects and demonstrates a more pronounced reduction in peak heart rate in those subjects compared with non hypoxemic ones. These data confirm the possible influence of arterial oxygenation on the decrease in peak heart rate in acute hypoxia. PMID- 17111316 TI - A cumulative effect of physical training on bone strength in males. AB - Weight-bearing, high-impact exercise, as opposed to nonimpact exercise, has been demonstrated to increase bone mineral density. This was traditionally demonstrated with dual energy X-ray absorptiometry. Our objective was to assess the differences in bone properties, using quantitative ultrasound (QUS, Sunlight Omnisense, Sunlight Medical, Ltd., Tel Aviv, Israel), in male athletes involved in a weight-bearing, impact sport (soccer, SC) or a nonimpact sport (swimming and water polo, AQ), compared with nonathletic control (C) males. A total of 266 boys and men, aged 8 - 23 years, were divided into children (11.1 +/- 1.0 years; 34 SC, 34 AQ, 25 C), adolescents (14.7 +/- 1.2 years; 32 SC, 31 AQ, 31 C), and young adults (19.8 +/- 1.1 years; 31 SC, 24 AQ, 24 C) . Training experience varied between 1.5 years in the children to 15 years in the adults. Bone speed of sound (SOS) was measured bilaterally at the distal radius and the mid-tibia. Body fat was significantly lower in athletes compared with C. AQ were generally heavier and had a higher fat-free mass compared with SC and C, with no significant differences in height between groups. Radial SOS increased with age, but no differences were observed between activity groups or between the dominant (D) and nondominant (ND) arm. Tibial SOS also increased with age. In the children and adolescents, no differences were observed between activity groups. However, among adults, both SC and AQ had higher tibial SOS compared with C. These differences were mainly explained by differences in fat-free mass. Among young adults but not among children and adolescent males, both soccer and aquatic sports appear to be associated with higher bone SOS in the lower, but not the upper, extremities. Further studies are needed to assess possible sport-specific mechanisms which affect bone properties and to determine the minimal cumulative effect which is needed to influence bone properties. PMID- 17111317 TI - Salivary cortisol, testosterone, and T/C ratio responses during a 36-hole golf competition. AB - The purpose of this investigation was to study the effects of 36 continuous holes of competitive golf on salivary testosterone, cortisol, and testosterone-to cortisol ratio and their relation to performance in eight elite male collegiate golfers (age 20.3 [+/- 1.5] years). Thirty-six holes of a 54-hole NCAA golf tournament were played on the first day of the competition. A saliva sample was taken 45 minutes prior to the round and immediately following each hole for a total of 37 samples per subject. Time matched baseline samples were collected on a different day to account for circadian variation. Six-hole areas under the curve (AUC) values were calculated for endocrine measures. Significant (p < 0.05) increases were noted for cortisol during competition, however, testosterone did not change during competition compared to baseline. Testosterone-to-cortisol (T/C) ratio was significantly lower throughout the competition compared to baseline measures. Thirty-six-hole AUC testosterone-to-cortisol ratio response was correlated (r = 0.82) to 36-hole score. There was a high correlation between pre-round testosterone (r = 0.71), T/C ratio response (r = 0.82), and 36-hole score. CSAI-2 somatic anxiety was correlated to pre-round cortisol (r = 0.81) and testosterone (r = - 0.80) response. These results indicate a significant hormonal response during 10 hours of competitive golf. Good golf performance (low golf scores) in this competition was related to low T/C ratio (r = .82). Additionally, results from this investigation validated CSAI-2 somatic anxiety with physiological measures of anxiety. PMID- 17111318 TI - Exercise associated genes in rat colon mucosa: upregulation of ornithin decarboxylase-1. AB - Epidemiology has revealed that physical activity is an important lifestyle factor that reduces the risk of colon cancer. However, the underlying mechanisms of this protective effect have so far not been defined. The aim of this study was to identify molecular targets of physical activity in rat colon mucosa by employing our voluntary exercise model. Twenty male rats underwent a 12-week exercise program, with 9 additional rats serving as a control group. Running distances, body weights and heart weights as measures of physical adaptations were recorded, and changes in mRNA steady-state levels of marker genes involved in vascularization (VEGF, HIF-1 alpha, ODC-1), apoptosis (Bcl-2, PPAR gamma) and prostaglandin synthesis (COX-2) were determined by qRT-PCR. The four housekeeping genes GAPDH, beta-actin, 18S and ALDA served as reference genes. Recorded running distances showed great inter-individual differences resulting in three different groups of low (L-EX, < 2629 m/night; n=5), moderate (M-EX, 3003 - 7458 m/night; n=10) and high (H-EX, > 8314 m/night; n=5) physical activity. The M-EX and H-EX group revealed significant (p<0.05) adaptive changes with an increase in heart mass per kg body weight and a decrease in mean body weight. Amongst the marker genes studied by mRNA expression analysis only ODC-1 appears to be differentially expressed. Its 1.8-fold increased steady-state mRNA level in the H-EX group suggests that synthesis of polyamines may be increased by physical activity. This new finding could provide a link between extensive physical activity and its protective effects on colon cancer development. PMID- 17111319 TI - Habitual physical activity, physical fitness and heart rate variability in preadolescents. AB - The amount and type of physical activity (PA) appropriate in youth for optimal health is still being discussed. The purpose of this study on 12-year-old preadolescents is to evaluate the association of two PA patterns, corresponding to different exercise intensities, with some health-related indexes. PA was evaluated by triaxial accelerometry in 67 student subjects, who were successively classified according to 1) their usual participation or not in moderately intense (> 4 metabolic equivalents, METs) activities (either in daily life or in leisure activities) for at least 210 min a week (4 M (210)); and 2) the habitual practice or not of more intense activities (> 6 METs) for at least 60 min a week (6 M (60)). Health-related indexes consisted of cardiopulmonary (maximum oxygen uptake, V.O 2max), muscular (lower limb strength) fitness parameters and vagal related indexes of heart rate (HR) variability (HRV), such as the HF/(LF+HF) ratio (where HF and LF stand for high and low frequency power). HR was recorded in the supine position after 15 min of quiet rest, and HRV indexes were calculated in 5-min segments. Energy expenditure due to PA was not significantly associated with any health-related index. Reaching 4 M (210) was associated with both higher estimated V.O 2max (p < 0.05) and higher muscular strength indexes (p < 0.05) but not with enhanced HRV indexes. Subjects who reached 6 M (60) had higher estimated V.O2max (p < 0.05) and higher HF/(LF+HF) (p < 0.05). Our results show that regular moderately intense activities are sufficient to influence physical fitness but suggest that more intense activities are necessary to observe more favorable HRV vagal-related indexes. PMID- 17111320 TI - Heart rate variability, training variation and performance in elite swimmers. AB - The aim of the present study was to investigate the relationships between heart rate variability (HRV) changes and both training variations and performances in elite swimmers. A secondary purpose was to measure catecholamine urinary excretion in elite swimmers to validate the HRV indices of sympathetic activity during training. Thirteen swimmers (4 females and 9 males) were tested before and after 4 weeks of intense training (IT) and 3 weeks of reduced training (RT). At the end of each period, the swimmers participated in an official competition of their best event. Individual performances were expressed as percentage of the previous season's best performance. Spectral analysis was used to investigate RR interval variability. HRV indices failed to show any significant changes between the study periods (p>0.05). Pre-IT HF was correlated with performance (r=0.45; p=0.05) and HFnu (r=0.59; p<0.05) during RT. On the other hand, once RT was completed, HFnu was correlated positively to performance (r=0.81; p<0.01) and negatively to fatigue (r=- 0.63; p<0.03). Conversely, the indices of sympathetic activity, i.e., LFnu and LF/HF ratio were inversely related to performance (both r=- 0.81; p<0.01); total fatigue score was correlated to the changes in HFnu (r=- 0.63; p<0.03) and in the LF/HF ratio (r=0.58; p<0.05). Changes in the adrenaline/noradrenaline ratio over the follow-up period were related to the changes in the LF/HF ratio (r=0.45; p<0.03). In highly trained swimmers coping well with a training program, including 4 weeks of IT followed by 3 weeks of RT, HRV indices were unaltered. On the other hand, after the 3 weeks of RT, HFnu was positively related to performance and inversely related to the fatigue score. Thus, elevated initial HF levels could be important in the parasympathetic activity increases during taper and, hence, in swimming performance improvement. PMID- 17111321 TI - Principal component structure and sport-specific differences in the running one leg vertical jump. AB - The aim of this study is to identify the kinetic principal components involved in one-leg running vertical jumps, as well as the potential differences between specialists from different sports. The sample was composed of 25 regional skilled athletes who play different jumping sports (volleyball players, handball players, basketball players, high jumpers and novices), who performed a running one-leg jump. A principal component analysis was performed on the data obtained from the 200 tested jumps in order to identify the principal components summarizing the six variables extracted from the force-time curve. Two principal components including six variables accounted for 78 % of the variance in jump height. Running one-leg vertical jump performance was predicted by a temporal component (that brings together impulse time, eccentric time and vertical displacement of the center of mass) and a force component (who brings together relative peak of force and power, and rate of force development). A comparison made among athletes revealed a temporal-prevailing profile for volleyball players, and a force dominant profile for Fosbury high jumpers. Novices showed an ineffective utilization of the force component, while handball and basketball players showed heterogeneous and neutral component profiles. Participants will use a jumping strategy in which variables related to either the magnitude or timing of force production will be closely coupled; athletes from different sporting backgrounds will use a jumping strategy that reflects the inherent demands of their chosen sport. PMID- 17111322 TI - Effects of detraining on the functional capacity of previously trained breast cancer survivors. AB - The purpose of this study was to assess the effects of a relatively short (8 weeks) period of detraining on cardiorespiratory capacity, dynamic strength endurance, task specific functional muscle capacity and quality of life (QOL) of breast cancer survivors who had previously undergone a combined supervised (aerobic and resistance) training program. Eleven women survivors of stage I - II ductal breast carcinoma (47 +/- 7 yrs) entered the study and performed a battery of tests (including anthropometric evaluation, a graded cycle ergometer test, tests of strength endurance [leg and bench press] and the sit-stand test) and completed a specific QOL questionnaire (EORTC-C30) at three time points: i) before, ii) after an exercise program (including aerobic and resistance exercises) of 8-weeks duration, and iii) after a subsequent 8-weeks period of training cessation. Training-induced improvements in strength endurance, muscle functional capacity (sit-stand test) and QOL were not significantly changed after detraining (p > 0.05 for post-training vs. detraining comparisons). The lack of significant loss in muscle strength endurance occurred despite significant losses in estimated total muscle mass after detraining (27.3 +/- 2.4 kg) compared with post-training (28.5 +/- 2.9 kg). In contrast, cardiorespiratory capacity was significantly decreased during detraining (V.O (2peak) of 29.0 +/- 4.6 vs. 22.7 +/- 3.9 ml . kg ( -1) . min (-1) at post-training vs. detraining, p < 0.01). In conclusion, cancer survivors who have participated in a combined training program can retain some of the training gains (particularly improved QOL and muscle strength endurance/functional performance) after a relatively short duration detraining period. PMID- 17111323 TI - Current physical activity is related to bone mineral density in males but not in females. AB - The purpose of the present study was to investigate the association between high , medium-, and low-impact physical activity in males and females at the time of peak bone mineral density in young adulthood. The cohort consisted of 62 male medical students (aged 28.1 +/- 3.9) and 62 female medical students (aged 25.1 +/ 3.9). The bone mineral density (aBMD, g/cm (2)) of the total body, femoral neck, and lumbar spine, and the bone mineral content (BMC, grams) and area (cm (2)) of the femoral neck and lumbar spine was measured using dual energy X-ray absorptiometry. Volumetric BMD (vBMD, mg/cm (3)) of the femoral neck and lumbar spine was estimated. The total amount of physical activity per week, which was recorded in a questionnaire, was divided into high-impact, medium-impact, and low impact activity. In the male cohort, hours of high-impact physical activity per week was associated with aBMD and BMC of all sites (r=0.27 - 0.53, p<0.05) and bone area of the femoral neck (r=0.38, p<0.01). Total amount of physical activity per week was associated with aBMD of the total body and femoral neck, BMC of femoral neck and lumbar spine, femoral neck vBMD, and the lumbar spine area (p<0.05 for all). Using multiple linear regression, high-impact physical activity was independently associated with aBMD (beta=0.27, p<0.05) and BMC (beta=0.34, p<0.01) of the femoral neck. In the female cohort there was no association between amount or type of physical activity to aBMD, BMC, vBMD, or the bone area of any site. Instead body weight, lean body mass, or fat mass were significantly related to aBMD and all BMC sites in this group. The results of the present study suggest that present physical activity level has a stronger relation to different aspects of bone mass in the male compared to the female adult skeleton. PMID- 17111324 TI - Barrett's esophagus. PMID- 17111325 TI - Endoscopic diagnosis and treatment of upper gastrointestinal tumors. PMID- 17111326 TI - Gastrointestinal bleeding. PMID- 17111327 TI - Enteroscopy and capsule endoscopy. PMID- 17111328 TI - Colon tumors and colonoscopy. PMID- 17111329 TI - ERCP topics. PMID- 17111330 TI - Endoscopic ultrasonography. PMID- 17111331 TI - Laparoscopy and combined methods. PMID- 17111332 TI - Confocal endomicroscopy for the diagnosis of gastric cancer in vivo. AB - BACKGROUND AND STUDY AIMS: Advances in endoscopy have led to imaging of the details of the gastric mucosa, but the histological diagnosis usually has to be confirmed by endoscopic biopsy. A method of confocal endomicroscopy that has recently been developed allows the observation of living cells in vivo. Several investigators have reported that the technique is of value, but there have as yet been no studies describing its application in gastric cancer. PATIENTS AND METHODS: Twenty-seven patients with early gastric cancer underwent confocal endomicroscopy (Pentax EG-3870CIK; Pentax, Tokyo, Japan). After intravenous administration of fluorescein sodium, confocal images obtained from the normal mucosa and from cancerous lesions were interpreted by two pathologists independently and compared with the histological findings, including CD34 immunostaining of biopsy specimens or resected specimens from the same sites. RESULTS: Fluorescein yielded high-quality confocal images of the gastric mucosa; if cancer could be targeted (59%) images were mostly graded good. The images corresponded to the hematoxylin-eosin staining of transverse sections of specimens from the same sites. In the results for the interpretation by the two pathologists, the accuracy for the diagnosis of gastric cancer was 94.2% (pathologist A), and 96.2% (pathologist B), respectively. The accuracy decreased substantially when poor images and inaccessible lesions were included. CONCLUSIONS: Confocal endomicroscopy is useful in the diagnosis of gastric cancer but good quality images cannot always be obtained. In the future, it may allow virtual biopsy and help reduce unnecessary biopsies. PMID- 17111333 TI - Ex-vivo study of high-magnification chromoendoscopy in the gastrointestinal tract to determine the optimal staining conditions for endocytoscopy. AB - BACKGROUND AND STUDY AIMS: Endocytoscopy allows the observation of living cells in the gastrointestinal tract. Consistently clear views are essential for clinical application of the technique, but these are not always obtained. The aim of this study was to determine an appropriate staining regimen for endocytoscopy. MATERIALS AND METHODS: This was an ex-vivo animal study in which we stained freshly resected porcine esophagus, stomach, and colon with different concentrations of three dyes (1%, 0.5%, and 0.25% crystal violet; 5%, 2.5%, and 1% methylene blue; and 1%, 0.5%, and 0.25% toluidine blue) and assessed them after different exposure times (10 seconds, 30 seconds, 60 seconds, and 90 seconds). The images obtained were evaluated according to the staining status of the cytoplasm and the nucleus, and the contrast between the cytoplasm and the nuclei, and the optimal staining conditions for each organ were determined. Additionally, freshly resected human esophagus, stomach, and colon tissues were tested under the dye/exposure conditions that were found to be the most appropriate in the animal study. RESULTS: After intensive mucus removal, high quality images were obtained using methylene blue and toluidine blue. The optimum conditions for endocytoscopic observation were obtained after staining with 1% methylene blue in the esophagus and with 0.25% toluidine blue in the stomach and the colon, after 60 seconds of exposure to the dye. This was confirmed in the human specimens. CONCLUSIONS: This study provides important information on appropriate staining conditions for endocytoscopy. Further ex-vivo and in-vivo studies are necessary before this technique comes into standard use, however. PMID- 17111334 TI - Delays in diagnosis and therapy of gastric cancer and esophageal adenocarcinoma. AB - BACKGROUND AND STUDY AIMS: In the past, there were long delays in the diagnosis of patients with cancer of the stomach or esophagus. The objective of this study was to describe current delays in the diagnosis and treatment of gastric and esophageal adenocarcinoma and to compare the findings with those from an historical control population treated at the same institutions 10 years earlier. PATIENTS AND METHODS: Patients with biopsy-proven gastric cancer or esophageal adenocarcinoma who were treated at two academic medical centers in Germany between April and October 2003 were consecutively screened for eligibility to take part in the study. Medical charts for each patient were reviewed. Additional data were obtained via structured interviews. Main outcome measures were the total delay, and the delays related to patients themselves, to doctors, and to the hospital. Data were compared with those from a historic control group assessed in 1993. RESULTS: The median total delay for patients with gastric cancer (n = 104) was 3.5 months (range 0.3 - 29.6), and in patients with esophageal adenocarcinoma (n = 22) the total delay was significantly shorter (median 2.2 months, range 1.2 - 11.7; P < 0.05). Comparing these findings with those from an historic cohort of patients with gastric cancer (n = 100) revealed a significant decrease in the total delay (3.5 versus 8.0 months, P < 0.001). CONCLUSIONS: The current findings indicate that delays in the diagnosis and treatment of gastric cancer have become significantly shorter within the last 10 years as our understanding of and ability to treat this form of cancer have improved. PMID- 17111335 TI - The role of wireless capsule endoscopy in investigating unexplained iron deficiency anemia after negative endoscopic evaluation of the upper and lower gastrointestinal tract. AB - INTRODUCTION: Despite undergoing standard endoscopic diagnostic evaluation with eosophagogastroduodenoscopy and ileocolonoscopy, up to 30% of patients with iron deficiency anemia (IDA) have no definitive diagnosis. The aim of this study was to prospectively investigate the role of wireless capsule endoscopy (WCE) in detecting lesions of the small bowel in patients with unexplained IDA after a negative endoscopic work-up. PATIENTS AND METHODS: Between 1 December 2003 and 31 December 2004, 253 consecutive patients who had been referred because of unexplained IDA underwent eosophagogastroduodenoscopy with small-bowel biopsies and ileocolonoscopy. Endoscopic and histological investigations were negative in 51 of these patients (20.2%) and WCE was performed. Air double-contrast enteroclysis was performed following WCE in all these patients. RESULTS: Wireless capsule endoscopy revealed one or more small-bowel lesions that were considered to be a likely cause of the IDA in 29/51 patients (57%): angiodysplasias in twelve patients (23.5%), multiple jejunal and/or ileal ulcers in six patients (11.7%), multiple erosions in four patients (7.8%), a solitary ulcer in three patients (5.9%), polyps in two patients (3.9%), and tumors in two patients (3.9%). Enteroclysis revealed abnormal findings likely to cause IDA in only 6/51 patients (11.8%): multiple ileal ulcers in three patients (5.9%), tumors in two patients (3.9%), and polyps in one patient (1.9%) (enteroclysis VS. WCE, P < 0.0001). WCE revealed all of the radiographic findings and no adverse events were observed. CONCLUSIONS: This study demonstrates the importance of investigating the small bowel with WCE in patients with unexplained IDA after negative standard endoscopic evaluation. Wireless capsule endoscopy is superior to enteroclysis for detecting lesions of the small bowel in patients with unexplained IDA and should be the next diagnostic test of choice after unremarkable standard endoscopic evaluation. PMID- 17111336 TI - Endoscopic retrograde cholangiopancreatography catheter and accessory exchange using a short hydrophilic guide wire: a prospective study. AB - BACKGROUND AND STUDY AIMS: Although hydrophilic guide wires can be used to facilitate stricture cannulation during endoscopic retrograde cholangiopancreatography (ERCP), some endoscopists avoid using them because of concerns about wire loss during exchange. There are no data available on the outcomes of using a short hydrophilic guide wire during ERCP. The aim of this study was to assess the outcomes of therapeutic ERCP procedures in which a short, completely hydrophilic guide wire was used exclusively. PATIENTS AND METHODS: A total of 100 patients undergoing ERCP were studied prospectively. A 0.035-inch, 260-cm long, angled-tip hydrophilic wire (Terumo Glidewire) was used initially. Hydraulic catheter exchange was performed as follows: during catheter withdrawal the assistant advanced all the available wire into the catheter; a 12-ml syringe was then attached to the catheter and water was flushed under pressure to "float the wire" and maintain its position during catheter removal. Variables evaluated included exchange times and wire loss rates. RESULTS: A total of 223 catheter exchanges were performed, 132 (59%) using the Olympus V-Scope (which held the wire in 62% of cases): 15% of exchanges were with catheters/accessories designed for short-wire use (Boston Scientific Rapid Exchange Biliary System or Rx System), and 85% were with a variety of standard-length accessories. Overall, the mean exchange time was 26 seconds (range 6 - 90 seconds, standard deviation 12 seconds). The mean exchange time was faster with the V-Scope and with non-Rx System accessories. Wire loss occurred in 5 % of all exchanges. Desired ductal/stricture access was achieved in all the patients. CONCLUSIONS: Exchange of short hydrophilic wires is quick and reliable. The Olympus V-Scope is able to hold the wire in some cases. Monorail-type devices and accessories slow catheter exchange down slightly because hydraulic exchange cannot be performed using these systems. PMID- 17111337 TI - Implementation of sedation guidelines in clinical practice in Italy: results of a prospective longitudinal multicenter study. AB - BACKGROUND AND STUDY AIMS: Following publication in 2000 of the Italian Guidelines for Sedation in Digestive Endoscopy, the authors carried out a dissemination program and an audit of the impact of the Guidelines in clinical practice. The primary aim was to evaluate any changes in sedation methods at the start of and during the study period (at 0, 6, and 12 months). Secondary study aims were detailed analyses of endoscopies regarding sedation, complications, and patient satisfaction. PATIENTS AND METHODS: The prospective, observational, multicenter study was commenced in 60 centers (31 first- and 29 second-level) which opted to have seminars on the Guidelines; results concerning sedation practices in relation to endoscopic procedure, complications, and patient satisfaction were recorded at the start of the study (phase 0) and at 6 and 12 months. Over a 1-year period, 5261 sedated and unsedated patients undergoing endoscopic procedures were enrolled (with data collection between December 1999 and June 2002). The results were analyzed using the chi-squared test. RESULTS: Data were available for all three assessment time points from 29 of the 60 centers. The sedation rate, between phase 0 and 2, increased by 12.75% in first level centers and 0.73% in second-level centers with midazolam use increasing from 34% to 41%. For colonoscopy, sedation usage increased from 52% to 56%. Adverse events occurred in 0.47% of the patients, without long-term sequelae or death, and patient satisfaction was only slightly, but not significantly higher with sedation (90.4% of sedated patients were "very satisfied" vs. 87% of unsedated patients who were "satisfied"), but the examination was more likely to be complete in sedated patients and this difference was statistically significant ( P < 0.001). CONCLUSIONS: We conclude that if sedation is used as recommended by the Guidelines, it is possible to control clinical risk by using high quality endoscopic procedures. PMID- 17111338 TI - From small-bowel bleeding to anemia: a wider range of indications for the capsule endoscope? PMID- 17111339 TI - Performance, length, and cost of endoscopic retrograde cholangiopancreatography guide wires: which is the best mix of features? PMID- 17111340 TI - Same-day endoscopic retrograde cholangiopancreatography after transduodenal endoscopic ultrasound-guided needle aspiration: do we need to be cautious? AB - Patients who are undergoing endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) for suspected pancreatic tumors frequently undergo endoscopic retrograde cholangiopancreatography (ERCP) for palliation of their symptoms. Performing EUS-FNA and ERCP in tandem may be cost-effective and may reduce procedure time, but the potential risks associated with this approach have not been clearly defined in the literature. We report two patients who underwent same day therapeutic ERCP after transduodenal EUS-FNA for pancreatic tumors. Endoscopic biliary manipulation during ERCP aggravated an inadvertent and subclinical needle puncture injury to the bile duct sustained during the preceding EUS-FNA. This resulted in leakage of bile into the retroperitoneal space, and both patients required laparotomy and surgical drainage. Additional clinical evidence is needed to clarify these issues and to determine whether it would be prudent to perform therapeutic ERCP prior to diagnostic transduodenal EUS-FNA when these two procedures are planned as sequential or same-day procedures. PMID- 17111341 TI - French Society of Digestive Endoscopy (SFED) Guidelines on performing a colonoscopy. PMID- 17111342 TI - [Polish diagnostic and therapeutic guidelines in gastroenteropancreatic neuroendocrine tumors (GEP NET)]. PMID- 17111344 TI - Prolonged exposure of dendritic cells to maturation stimuli favors the induction of type-2 cytotoxic T lymphocytes. AB - Dendritic cell (DC) maturation influences the priming and polarization of T lymphocytes. We recently found that early activated DC (i.e. DC exposed to pro maturation stimuli for 8 h) were more prone to prime in vivo a type-1 cytotoxic T cell (Tc1) response than DC exposed to pro-maturation stimuli for 48 h (48h-DC). We investigated whether 48h-DC, conversely, allowed the induction of Tc2 cells. Antigen-pulsed mouse bone-marrow-derived DC at any maturation stage, in the presence of exogenous IL-12, skewed in vitro naive CD8(+) T cells towards Tc1 cells, but 48h-DC most potently, in the presence of exogenous IL-4, favored the induction of Tc2 cells. In vivo, full maturation of DC promoted expansion of Tc2 and fall of Tc1 cells. Tc2 cells maintained a high cytolytic activity and produced significant amounts of IL-4, IL-5, IL-10 and TGF-beta. Our results indicate that polarization of naive CD8(+) T cells to Tc2 cells is dependent on the amount of time DC have been exposed to maturation stimuli, and might be favored in late and/or chronic phases of an immune response. PMID- 17111345 TI - Expression and role of phosphatidylcholine-specific phospholipase C in human NK and T lymphocyte subsets. AB - We recently reported evidence of phosphatidylcholine-specific phospholipase C (PC PLC) involvement in NK cell-mediated cytotoxicity and in lytic granule exocytosis. In the present study, different subpopulations of human PBL were investigated in relation to PC-PLC enzyme expression. While a substantial intracellular amount of PC-PLC was detected in all lymphoid subsets, expression of this enzyme on the outer membrane surface reached high levels only in NK cells, was present at low levels in B lymphocytes and in some TCR gamma/delta T cells and was practically absent in CD4(+) and CD8(+ )T lymphocytes. Moreover, in NK cells two different subpopulations were identified, CD56(dim) PC-PLC(bright) and CD56(bright) PC-PLC(low/-) cells, corresponding to distinct subsets with cytolytic and immunoregulatory functions, respectively. Interestingly, the PC-PLC expression level on the NK membrane surface correlated closely with that of the CD16 receptor, suggesting a possible relationship between enzyme externalization and NK cell maturation. In summary, our results suggest that a high PC-PLC expression on the cell membrane surface of PBL is a peculiarity of NK cytolytic cells, in which the enzyme is apparently involved in the ability of this subset to lyse sensitive target cells. PMID- 17111346 TI - Ions in space. AB - We review the detection history, observation, distribution, and reactivity of molecular ions in extraterrestrial space, with particular (though not exclusive) reference to interstellar monocations. The diversity of interstellar ion chemistry is highlighted with reaction examples, drawn from the authors' own laboratories and elsewhere, and attempt to provide an overview of this broad and increasingly divergent field. Emphasis is given to the role of ions in the synthesis of molecules, including their ability to catalyze the transformation of neutral molecules. PMID- 17111347 TI - Nucleic acid agonists for Toll-like receptor 7 are defined by the presence of uridine ribonucleotides. AB - Toll-like receptor 7 (TLR7) mediates innate responses by responding to viral RNA in endocytic compartments. However, the molecular pattern recognised by TLR7 and whether it differs between RNA of viral and self origin remains unclear. Here, we identify nucleic acids that act as TLR7 agonists for mouse and human cells. We show that uridine and ribose, the two defining features of RNA, are both necessary and sufficient for TLR7 stimulation, and that short single-stranded RNA (ssRNA) act as TLR7 agonists in a sequence-independent manner as long as they contain several uridines in close proximity. Consistent with the notion that TLR7 lacks specificity for sequence motifs, we show that it is triggered equally efficiently by viral or self RNA delivered to endosomes. Our results support the notion that TLR7 recognises uracil repeats in RNA and that it discriminates between viral and self ligands on the basis of endosomal accessibility rather than sequence. PMID- 17111348 TI - Interleukin-10 derived from macrophages and/or neutrophils regulates the inflammatory response to LPS but not the response to CpG DNA. AB - Interleukin-10 (IL-10) is an important regulator of immune responses secreted by different cell types. We have previously shown that mice with selective inactivation of the IL-10 gene in T cells suffer from deregulated T cell responses similar to those observed in IL-10(-/-) animals. Unlike IL-10(-/-) mice, however, T cell-specific mutants do not mount an enhanced innate immune response to LPS, which must, therefore, be subject to control by IL-10 from non-T cells. Herein we show that subcutaneous injection of LPS, which causes moderate local inflammation in WT and T cell-specific IL-10 mutant mice, results in augmented inflammatory infiltration and extensive tissue necrosis in mice with deficiency for IL-10 in macrophages and neutrophils. Correspondingly, we observed an enhanced sensitivity of the macrophage/neutrophil-specific IL-10 mutants to systemic LPS exposure when compared with WT animals. In contrast, the inflammatory response of these mutants to CpG oligodeoxynucleotides was not different from that of WT mice. While IL-10(-/-) mice developed massive inflammation, necrosis and increased serum cytokine levels after subcutaneous CpG injection, only moderate responses were observed in macrophage/neutrophil specific IL-10 mutant and WT mice. These results show that different innate immune responses can be subject to control by IL-10 from different cellular sources. PMID- 17111349 TI - Structure and duration of contact between dendritic cells and T cells are controlled by T cell activation state. AB - The adaptive immune response is initiated when naive T cells interact with dendritic cells (DC). However, the physicodynamics as well as the molecules that constitute the contact plane (immunological synapse) between DC and T cells are not well understood. We show here that for the formation of stable conjugates, T cells need to be preactivated by DC in a CD80/86- and antigen dose-dependent manner. When activated, T cells induce cytoskeletal reorganization within DC via CD40-CD40L signaling. Polarization of the actin and fascin cytoskeleton in DC is associated with sustained DC-T cell contacts, strong T cell proliferation and a Th1 response. Organized contact planes with clearly separated patches containing TCR or CD11a are also formed. Thus, DC-T cell interactions take place in a sequential, interdependent fashion: first, DC "license" naive T cells to engage DC in an antigen dose- and CD80/86-dependent fashion. Then, these preactivated T cells induce cytoskeletal reorientation in DC, resulting in sustained DC-T cell contacts and subsequent T cell activation. These results demonstrate that T cells control the mode of interaction based on information gathered from DC. PMID- 17111350 TI - Regulation of 2B4 (CD244)-mediated NK cell activation by ligand-induced receptor modulation. AB - Natural killer (NK) cell activity can be stimulated by different surface receptors. 2B4 is a member of the signaling lymphocyte activation molecule (SLAM) related receptor family and is important for stimulating human NK cell cytotoxicity and cytokine production. Here we show that stimulation of human NK cells by antibody-mediated 2B4 cross-linking or incubation with target cells expressing the 2B4 ligand CD48 results in a strong down-modulation of 2B4 surface expression. This down-modulation is observed in NK cell lines, purified human NK cells and NK cell clones, and is accompanied by an internalization of 2B4. The modulation of 2B4 is dependent on the activity of Src-family kinases, but independent of PI3 K activity or actin polymerization. Inhibitory receptors can interfere with 2B4-mediated signals and NK cell activation. However, co engagement of inhibitory killer cell Ig-like receptors has no influence on the down-modulation of 2B4. This suggests that the modulation of 2B4 expression is independent of inhibitory receptors. The lower surface expression of 2B4 after ligand-induced down-modulation results in reduced 2B4-mediated NK cell activation and cytotoxicity. The modulation of activating surface receptors may therefore be another mechanism for the fine-tuning of NK cell activity and may lead to the adaptation of NK cell cytotoxicity in tissues with high ligand expression. PMID- 17111351 TI - Alpha 2,3-sialyltransferase-IV is essential for L-selectin ligand function in inflammation. AB - L-selectin belongs to the C-type lectin family of glycoproteins and is constitutively expressed on most leukocytes. L-selectin mediates leukocyte rolling in inflamed microvessels and high endothelial venules (HEV) via binding to specific carbohydrate structures on selectin ligands. Previous studies using sialidase treatment suggested a role of sialic acid residues in L-selectin dependent rolling. To investigate the role of the alpha2,3-sialyltransferase (ST3Gal)-IV on L-selectin ligand activity in vivo, we studied leukocyte rolling in inflamed venules of the cremaster muscle and in Peyer's patch HEV of ST3Gal-IV deficient mice and littermate control mice. In cremaster muscle venules with or without TNF-alpha treatment, L-selectin-dependent rolling was almost completely abolished in ST3Gal-IV(-/-) mice. In both models, L-selectin interacts with P selectin glycoprotein ligand-1 (PSGL-1) presented by adherent leukocytes and leukocyte fragments, but not with endothelial L-selectin ligands. In contrast, L selectin-dependent rolling in Peyer's patch HEV, which is mediated by unknown endothelial L-selectin ligands, was not impaired in the absence of ST3Gal-IV. Our in vivo data show that PSGL-1, the molecule responsible for L-selectin-mediated leukocyte interactions in inflammation, is dependent on ST3Gal-IV, while alpha2,3 sialylation by ST3Gal-IV is not necessary for L-selectin ligand activity on high endothelial cells of Peyer's patch HEV. PMID- 17111352 TI - Temporal cross-talk between TCR and STAT signals for CD8 T cell effector differentiation. AB - The strength and duration of signaling through surface receptors is a primary means of controlling cell fate decisions. In adaptive immunity, Ag-initiated T cell stimulation is secondarily regulated by cytokines. We here summarize evidence for temporal control of a gene expression program in naive CD8 T cells. It is initiated in response to TCR engagement but relies on secondary signaling from cytokine receptors to be sustained and to allow development of full effector capacity. This mechanism permits cytokine receptor signaling to rescue abortive TCR signaling, such as that induced in response to weak or partial TCR agonists. Indeed, limiting TCR-initiated signaling on the Ras/ERK pathway may be complemented by STAT activation. Thus, TCR- and cytokine-driven activation of transcription factors and epigenetic modifications may act in concert in a temporally staggered process to establish the functional program of effector CD8 T cells. Based on gene expression profiling, molecular targets whose activation or inactivation may boost or dampen CD8 T cell effectors are also identified. Manipulation of these targets may, respectively, increase anti-tumor responses or prevent graft-versus-host reactions. PMID- 17111353 TI - Transient Notch signaling induces NK cell potential in Pax5-deficient pro-B cells. AB - Unlike early B/T cell development, NK cell lineage commitment is not well understood, with a major limitation being the lack of a robust culture system to assay NK cell progenitors. Here we have exploited the multi-lineage potential of Pax5(-/-) pro-B cells to establish an effective system to direct differentiation of progenitors into the NK cell lineage. Cultivation of Pax5(-/-) pro-B cells on OP9 cells expressing the Notch ligand Delta-Like1 (OP9-DL1) in the presence of IL 7 efficiently induced T and NK cell potential. For NK cells, Notch was only transiently required, as prolonged signaling decreased NK and increased T cell development. Pure NK cell populations could be obtained by the culture of these Notch signal-experienced cells onto OP9 stroma and IL-15. A similar transient exposure to Notch was also compatible with the differentiation of NK cells from hematopoietic progenitors, while sustained Notch signaling impaired NK cell generation. Pax5(-/-) pro-B cell-derived NK cells were cytotoxic, secreted cytokines and expressed all the expected NK cell-specific surface markers examined except the Ly49 family, a phenotype similar to fetal NK cells. These data indicate that Notch signaling induces T/NK cell differentiation in Pax5(-/-) pro-B cells that is strikingly similar to early thymopoiesis. PMID- 17111354 TI - Involvement of GATA3 in protein kinase C theta-induced Th2 cytokine expression. AB - Protein kinase C theta (PKCtheta) is essential for T cell activation, as it is required for the activation of NF-kappaB and expression of IL-2. PKCtheta has also been shown to affect NFAT activation and Th2 differentiation. To better understand the role of PKCtheta in the regulation of T helper cells, we used PKCtheta-deficient DO11.10 transgenic T cells to study its role in vitro. DO11.10 Th1 cells deficient in PKCtheta produced significantly less TNF-alpha and IL-2. The expression of Th2 cytokines, including IL-4, IL-5, IL-10, IL-13 and IL-24 was significantly reduced in PKCtheta-deficient T cells. Moreover, the expression of the Th2 transcription factor, GATA3, was significantly reduced in PKCtheta deficient T cells. Overexpression of GATA3 by retroviral infection in PKCtheta deficient T cells resulted in increased expansion of IL-4-producing T cells and higher IL-4 production than that of wild type Th2 cells. IL-5, IL-10, IL-13 and IL-24 expressions were also rescued by GATA3 overexpression. Our observations suggest that PKCtheta regulates Th2 cytokine expression via GATA3. PMID- 17111355 TI - Genomic suppression of murine B29/Ig-beta promoter-driven transgenes. AB - Immunoglobulin beta (Ig-beta) is a critical signal transducer of precursor B cell and B cell receptors. B29, the gene coding for Ig-beta, is switched on in progenitor B cells and expressed until the terminal stage of antibody-producing plasma cells. Although several cis-acting elements and transcription factors required for B29 expression have been characterized in cell lines, the in vivo significance of individual motifs located in the 1.2-kb promoter region remained unclear. To address whether this region drives B lineage-specific expression in mice as efficiently as in transfected cell lines, we established transgenic animals carrying the B29 promoter fused to either enhanced green fluorescent protein (EGFP) or the precursor B cell receptor component lambda5. Surprisingly, only minimal levels of B29-derived transcripts were produced in B lymphoid tissues of several independent transgenic lines, and the respective proteins were below the detection limit. In addition, transgenic transcripts were found in testis, kidney and brain. Hence, the 1.2-kb-sized B29 promoter does not define a strong, B lineage-restricted expression unit when randomly integrated into the genome and passed through the murine germ line. Therefore, yet unidentified genomic locus control elements are required to efficiently drive B29 expression in B lymphocytes. PMID- 17111356 TI - Cell cycle-dependent activity of the volume- and Ca2+-activated anion currents in Ehrlich lettre ascites cells. AB - Recent evidence implicates the volume-regulated anion current (VRAC) and other anion currents in control or modulation of cell cycle progression; however, the precise involvement of anion channels in this process is unclear. Here, Cl- currents in Ehrlich Lettre Ascites (ELA) cells were monitored during cell cycle progression, under three conditions: (i) after osmotic swelling (i.e., VRAC), (ii) after an increase in the free intracellular Ca2+ concentration (i.e., the Ca2+-activated Cl- current, CaCC), and (iii) under steady-state isotonic conditions. The maximal swelling-activated VRAC current decreased in G1 and increased in early S phase, compared to that in G0. The isotonic steady-state current, which seems to be predominantly VRAC, also decreased in G1, and increased again in early S phase, to a level similar to that in G0. In contrast, the maximal CaCC current (500 nM free Ca2+ in the pipette), was unaltered from G0 to G1, but decreased in early S phase. A novel high-affinity anion channel inhibitor, the acidic di-aryl-urea NS3728, which inhibited both VRAC and CaCC, attenuated ELA cell growth, suggesting a possible mechanistic link between cell cycle progression and cell cycle-dependent changes in the capacity for conductive Cl- transport. It is suggested that in ELA cells, entrance into the S phase requires an increase in VRAC activity and/or an increased potential for regulatory volume decrease (RVD), and at the same time a decrease in CaCC magnitude. PMID- 17111357 TI - Patterning the developing and regenerating olfactory system. AB - The olfactory system is a remarkable model for investigating the factors that influence the guidance of sensory axon populations to specific targets in the CNS. Since the initial discovery of the vast odorant receptor (ORs) gene family in rodents and the subsequent finding that these molecules directly influence targeting, several additional olfactory axon guidance cues have been identified. Two of these, ephrins and semaphorins, have well-established functions in patterning axon connections in other systems. In addition, lactosamine-containing glycans are also required for proper targeting and maintenance of olfactory axons, and may also function in other sensory regions. It is now apparent that these and likely other additional molecules are required along with ORs to orchestrate the complex pattern of convergence and divergence that is unique to the olfactory system. PMID- 17111358 TI - Neural specialization for hovering in hummingbirds: hypertrophy of the pretectal nucleus Lentiformis mesencephali. AB - Hummingbirds possess an array of morphological and physiological specializations that allow them hover such that they maintain a stable position in space for extended periods. Among birds, this sustained hovering is unique to hummingbirds, but possible neural specializations underlying this behavior have not been investigated. The optokinetic response (OKR) is one of several behaviors that facilitates stabilization. In birds, the OKR is generated by the nucleus of the basal optic root (nBOR) and pretectal nucleus lentiformis mesencephali (LM). Because stabilization during hovering is dependent on the OKR, we predicted that nBOR and LM would be significantly enlarged in hummingbirds. We examined the relative size of nBOR, LM, and other visual nuclei of 37 species of birds from 13 orders, including nine hummingbird species. Also included were three species that hover for short periods of time (transient hoverers; a kingfisher, a kestrel, and a nectarivorous songbird). Our results demonstrate that, relative to brain volume, LM is significantly hypertrophied in hummingbirds compared with other birds. In the transient hoverers, there is a moderate enlargement of the LM, but not to the extent found in the hummingbirds. The same degree of hypertrophy is not, however, present in nBOR or the other visual nuclei measured: nucleus geniculatus lateralis, pars ventralis, and optic tectum. This selective hypertrophy of LM and not other visual nuclei suggests that the direction selective optokinetic neurons in LM are critical for sustained hovering flight because of their prominent role in the OKR and gaze stabilization. PMID- 17111359 TI - Magnetic resonance imaging and histological studies of corpus callosal and hippocampal abnormalities linked to doublecortin deficiency. AB - Mutated doublecortin (DCX) gives rise to severe abnormalities in human cortical development. Adult Dcx knockout mice show no major neocortical defects but do have a disorganized hippocampus. We report here the developmental basis of these hippocampal abnormalities. A heterotopic band of neurons was identified starting at E17.5 in the CA3 region and progressing throughout the CA1 region by E18.5. At neonatal stages, the CA1 heterotopic band was reduced, but the CA3 band remained unchanged, continuing into adulthood. Thus, in mouse, migration of CA3 neurons is arrested during development, whereas CA1 cell migration is retarded. On the Sv129Pas background, magnetic resonance imaging (MRI) also suggested abnormal dorsal hippocampal morphology, displaced laterally and sometimes rostrally and associated with medial brain structure abnormalities. MRI and cryosectioning showed agenesis of the corpus callosum in Dcx knockout mice on this background and an intermediate, partial agenesis in heterozygote mice. Wild-type littermates showed no callosal abnormalities. Hippocampal and corpus callosal abnormalities were also characterized in DCX-mutated human patients. Severe hippocampal hypoplasia was identified along with variable corpus callosal defects ranging from total agenesis to an abnormally thick or thin callosum. Our data in the mouse, identifying roles for Dcx in hippocampal and corpus callosal development, might suggest intrinsic roles for human DCX in the development of these structures. PMID- 17111360 TI - Callosal axon arbors in the limb representations of the somatosensory cortex (SI) in the agouti (Dasyprocta primnolopha). AB - The present report compares the morphology of callosal axon arbors projecting from and to the hind- or forelimb representations in the primary somatosensory cortex (SI) of the agouti (Dasyprocta primnolopha), a large, lisencephlic Brazilian rodent that uses forelimb coordination for feeding. Callosal axons were labeled after single pressure (n = 6) or iontophoretic injections (n = 2) of the neuronal tracer biotinylated dextran amine (BDA, 10 kD), either into the hind- (n = 4) or forelimb (n = 4) representations of SI, as identified by electrophysiological recording. Sixty-nine labeled axon fragments located across all layers of contralateral SI representations of the hindlimb (n = 35) and forelimb (n = 34) were analyzed. Quantitative morphometric features such as densities of branching points and boutons, segments length, branching angles, and terminal field areas were measured. Cluster analysis of these values revealed the existence of two types of axon terminals: Type I (46.4%), less branched and more widespread, and Type II (53.6%), more branched and compact. Both axon types were asymmetrically distributed; Type I axonal fragments being more frequent in hindlimb (71.9%) vs. forelimb (28.13%) representation, while most of Type II axonal arbors were found in the forelimb representation (67.56%). We concluded that the sets of callosal axon connecting fore- and hindlimb regions in SI are morphometrically distinct from each other. As callosal projections in somatosensory and motor cortices seem to be essential for bimanual interaction, we suggest that the morphological specialization of callosal axons in SI of the agouti may be correlated with this particular function. PMID- 17111361 TI - Proinflammatory cytokine synthesis in the injured mouse spinal cord: multiphasic expression pattern and identification of the cell types involved. AB - We have studied the spatial and temporal distribution of six proinflammatory cytokines and identified their cellular source in a clinically relevant model of spinal cord injury (SCI). Our findings show that interleukin-1beta (IL-1beta) and tumor necrosis factor (TNF) are rapidly (<5 and 15 minutes, respectively) and transiently expressed in mice following contusion. At 30-45 minutes post SCI, IL 1beta and TNF-positive cells could already be seen over the entire spinal cord segment analyzed. Multilabeling analyses revealed that microglia and astrocytes were the two major sources of IL-1beta and TNF at these times, suggesting a role for these cytokines in gliosis. Results obtained from SCI mice previously transplanted with green fluorescent protein (GFP)-expressing hematopoietic stem cells confirmed that neural cells were responsible for the production of IL-1beta and TNF for time points preceding 3 hours. From 3 hours up to 24 hours, IL-1beta, TNF, IL-6, and leukemia inhibitory factor (LIF) were strongly upregulated within and immediately around the contused area. Colocalization studies revealed that all populations of central nervous system resident cells, including neurons, synthesized cytokines between 3 and 24 hours post SCI. However, work done with SCI-GFP chimeric mice revealed that at least some infiltrating leukocytes were responsible for cytokine production from 12 hours on. By 2 days post-SCI, mRNA signal for all the above cytokines had nearly disappeared. Notably, we also observed another wave of expression for IL-1beta and TNF at 14 days. Overall, these results indicate that following SCI, all classes of neural cells initially contribute to the organization of inflammation, whereas recruited immune cells mostly contribute to its maintenance at later time points. PMID- 17111362 TI - PECAM-1 modulates thrombin-induced tissue factor expression on endothelial cells. AB - Platelet endothelial cell adhesion molecule-1 (PECAM-1) (CD31) is known to inhibit platelet function and thrombus formation. The mechanisms involved in PECAM-1's roles as a modulator of hemostasis are still not completely understood. We examined the role of PECAM-1 as a regulator of tissue factor (TF) expression, a known important inducer of thrombosis. Wildtype and CD31KO mice underwent transient (30 min) renal ischemia followed by 24 h re-perfusion and their kidneys assessed for apoptosis, fibrin formation, and tissue factor expression. CD31KO mice exhibited increased tubular epithelial and endothelial apoptosis, increased fibrin deposition, and tissue factor expression. Human umbilical vein endothelial cells (HUVEC) transfected with antisense (AS) PECAM-1 oligonucleotides to downregulate PECAM-1 expression, exhibited greater induction of TF mRNA and protein expression as well as increased expression and nuclear localization of the transcription factor Egr-1 compared to scrambled AS PECAM-1 (Scr)-treated HUVEC following thrombin stimulation. TF induction was found to be mediated through thrombin receptor PAR-1 and the Galphai/o subunit of G-protein, confirmed by PAR-1 antagonist and pertussis toxin inhibition respectively. Thrombin mediated TF induction was dependent on Rho Kinase activity, phosphorylation of p38(MAPK) and p85 & Akt dephosphorylation. The inverse correlation of PI3K-Akt phosphorylation with p38 (MAPK) phosphorylation was confirmed by pharmacological inhibition. These studies suggest that PECAM-1 is involved in regulating a signaling pathway, affecting PI3K and Akt activation, p38 (MAPK) phosphorylation, which in turn, affects Egr-1 expression and nuclear translocation, ultimately affecting TF expression. These findings provide new insights into the action of PECAM-1 as a modulator of thrombosis. PMID- 17111363 TI - HIV-1 negatively affects the survival/maturation of cord blood CD34(+) hematopoietic progenitor cells differentiated towards megakaryocytic lineage by HIV-1 gp120/CD4 membrane interaction. AB - To investigate the mechanisms involved in the human immunodeficiency virus type 1 (HIV-1)-related thrombocytopenia (TP), human umbilical cord blood (UCB) CD34(+) hematopoietic progenitor cells (HPCs) were challenged with HIV-1(IIIb) and then differentiated by thrombopoietin (TPO) towards megakaryocytic lineage. This study showed that HIV-1, heat-inactivated HIV-1, and HIV-1 recombinant gp120 (rgp120) activated apoptotic process of megakaryocyte (MK) progenitors/precursors and decreased higher ploidy MK cell fraction. All these inhibitory effects on MK survival/maturation and platelets formation were elicited by the interaction between gp120 and CD4 receptor on the cell membrane in the absence of HIV-1 productive infection. In fact, in our experimental conditions, HPCs were resistant to HIV-1 infection and no detectable productive infection was observed. We also evaluated whether the expression of specific cytokines, such as TGF-beta1 and APRIL, involved in the regulation of HPCs and MKs proliferation, was modulated by HIV-1. The specific protein and mRNA detection analysis, during TPO induced differentiation, demonstrated that HIV-1 upregulates TGF-beta1 and downregulates APRIL expression through the CD4 engagement by gp120. Altogether, these data suggest that survival/differentiation of HPCs committed to MK lineage is negatively affected by HIV-1 gp120/CD4 interaction. This long-term inhibitory effect is also correlated to specific cytokines regulation and it may represent an additional mechanism to explain the TP occurring in HIV-1 patients. PMID- 17111364 TI - Connective tissue growth factor (CTGF) acts as a downstream mediator of TGF-beta1 to induce mesenchymal cell condensation. AB - Mesenchymal cell (MC) condensation or the aggregation of MCs precedes chondrocyte differentiation and is required for subsequent cartilage formation during endochondral ossification. In this study, we used micromass cultures of C3H10T1/2 cells as an in vitro model system for studying MC condensation and the events important for this process. Transforming growth factor beta1 (TGF-beta1) served as the initiator of MC condensation in our model system and we were interested in determining whether CTGF functions as a downstream mediator of TGF-beta1. CTGF is a matricellular protein that has been found to be expressed in MC condensations and in the perichondrium. Micromass cultures of C3H10T1/2 cells condensed under TGF-beta1 stimulation concomitant with dramatic up-regulation of CTGF mRNA and protein levels. CTGF silencing by either CTGF siRNA or CTGF antisense oligonucleotide approaches showed that TGF-beta1-induced condensation was CTGF dependent. Furthermore, silencing of CTGF expression resulted in significant reductions in cell proliferation and migration, events that are crucial during MC condensation. In addition, up-regulation of Fibronectin (FN) and suppression of Sox9 expression by TGF-beta1 was also found to be mediated by CTGF. Immunofluorescence of developing mouse vertebrae showed that CTGF, TGF-beta1 and FN were co-expressed in condensations of MCs, while Sox9 expression was low at this stage. During subsequent chondrogenesis, Sox9 expression was high in chondrocytes while CTGF expression was limited to the perichondrium. Thus, CTGF is an essential downstream mediator of TGF-beta1-induced MC condensation through its effects on cell proliferation and migration. CTGF is also involved in up regulating FN and suppressing Sox9 expression during TGF-beta1 induced MC condensation. PMID- 17111365 TI - Activation of the beta myosin heavy chain promoter by MEF-2D, MyoD, p300, and the calcineurin/NFATc1 pathway. AB - Calcium is a key element in intracellular signaling in skeletal muscle. Changes in intracellular calcium levels are thought to mediate the fast-to-slow transformation of muscle fiber type. One factor implicated in gene regulation in adult muscle is the nuclear factor of activated T-cells (NFAT) isoform c1, whose dephosphorylation by the calcium/calmodulin-dependent phosphatase calcineurin facilitates its nuclear translocation. Here, we report that differentiated C2C12 myotubes predominantly expressing fast-type MyHCII protein undergo fast-to-slow transformation following calcium-ionophore treatment, with several transcription factors and a transcriptional coactivator acting in concert to upregulate the slow myosin heavy chain (MyHC) beta promoter. Transient transfection assays demonstrated that the calcineurin/NFATc1 signaling pathway is essential for MyHCbeta promoter activation during transformation of C2C12 myotubes but is not sufficient for complete fast MyHCIId/x promoter inhibition. Along with NFATc1, myocyte enhancer factor-2D (MEF-2D) and the myogenic transcription factor MyoD transactivated the MyHCbeta promoter in calcium-ionophore-treated myotubes in a calcineurin-dependent manner. To elucidate the mechanism involved in regulating MyHCbeta gene expression, we analyzed the -2.4-kb MyHCbeta promoter construct for cis-regulatory elements. Using electrophoretic mobility shift assays (EMSAs), chromatin immunoprecipitation assays (ChIP), and nuclear complex coimmunoprecipitation (NCcoIP) assays, we demonstrated calcium-ionophore-induced binding of NFATc1 to a NFAT consensus site adjacent to a MyoD-binding E-box. At their respective binding sites, both NFATc1 and MyoD recruited the transcriptional coactivator p300, and in turn, MEF-2D bound to the MyoD complex. The calcium-ionophore-induced effects on the MyHCbeta promoter were shown to be calcineurin-dependent. Together, our findings demonstrate calcium-ionophore induced activation of the beta MyHC promoter by NFATc1, MyoD, MEF-2D, and p300 in a calcineurin-dependent manner. PMID- 17111366 TI - Differentiation of human adult skin-derived neuronal precursors into mature neurons. AB - The isolation of autologous neuronal precursors from skin-derived precursor cells extracted from adult human skin would be a very efficient source of neurons for the treatment of various neurodegenerative diseases. The purpose of this study was to demonstrate that these neuronal precursors were able to differentiate into mature neurons. We isolated neuronal precursors from breast skin and expanded them in vitro for over ten passages. We showed that 48% of these cells were proliferating after the first passage, while this growth rate decreased after the second passage. We demonstrated that 70% of these cells were nestin-positive after the third passage, while only 17% were neurofilament M-positive after 7 days of differentiation. These neuronal precursors expressed betaIII tubulin, the dendritic marker MAP2 and the presynaptic marker synaptophysin after 7 days of in vitro maturation. They also expressed the postsynaptic marker PSD95 and the late neuronal markers NeuN and neurofilament H after 21 days of differentiation, demonstrating they became terminally differentiated neurons. These markers were still expressed after 50 days of culture. The generation of autologous neurons from an accessible adult human source opens many potential therapeutic applications and has a great potential for the development of experimental studies on normal human neurons. PMID- 17111367 TI - N-glycosylation of the Xenopus laevis ClC-5 protein plays a role in cell surface expression, affecting transport activity at the plasma membrane. AB - Mutations in the gene encoding ClC-5 lead to X-linked hypercalciuric nephrolithiasis (XLHN), characterized by proteinuria, hypercalciuria, and phosphaturia. In renal proximal tubule cells, ClC-5 was identified as an important player in endocytosis, which ensures reabsorption of filtered protein. However, the recent finding that ClC-5 is a Cl(-)/H(+) antiporter and not a Cl(-) channel as long thought points to the lack of understanding of its functional role. Also, little biochemical data are available about ClC-5 and its post translational modifications have not been investigated. Here, we examined the role of N-glycosylation of xClC-5 in the Xenopus oocyte expression system by comparing wild-type (WT) xClC-5 and N-glycosylation site mutants. We found that xClC-5 is N-glycosylated on asparagines 169 and 470, which are the only N glycosylated sites. xClC-5 mutants have an increased susceptibility to polyubiquitination and proteasomal degradation; however, without a notable impact on the expression level. Using a cross-linking reagent, we showed that xClC-5 assembles into protein complexes, independent of its N-glycosylation. Voltage clamp measurements showed a reduced conductance in the presence of tunicamycin and with xClC-5 N-glycosylation site mutants. Using immunocytochemistry, we localized xClC-5 mainly in intracellular compartments, and found that its cell surface pool is reduced in the absence of N-glycans. We further examined the plasma membrane retrieval of WT and mutant xClC-5 in the presence of Brefeldin A (BFA), and found that the non-glycosylated mutant was retrieved more than five times faster than the WT protein. We conclude that N-glycosylation enhances cell surface expression of xClC-5, increasing its plasma membrane transport activity. PMID- 17111368 TI - Solitary gastric melanotic schwannoma: sonographic findings. AB - Solitary gastric schwannoma is rare, and solitary melanotic schwannoma is even rarer, posing a dilemma in diagnosis and treatment. We report the case of a 69 year-old woman with gastric melanotic schwannoma who presented with nausea, vomiting, and abdominal pain. Abdominal sonographic examination revealed a 5-cm hypoechoic mass in the epigastric area that was confirmed to be a gastric submucosal tumor on endoscopic examination. The diagnosis of melanotic schwannoma was confirmed via sonographically guided percutaneous core biopsy. The tumor was resected, and no recurrence has occurred in a 3-year follow-up. PMID- 17111370 TI - Combinatorial treatment of non-small-cell lung cancers with gefitinib and Ad.mda 7 enhances apoptosis-induction and reverses resistance to a single therapy. AB - Activation of the epidermal growth factor receptor (EGFR) contributes to the pathogenesis of non-small-cell lung carcinomas (NSCLC) and gefitinib, a selective reversible EGFR inhibitor, is effective in treating patients with NSCLC. However, clinical resistance to gefitinib is a frequent occurrence highlighting the need for improved therapeutic strategies. Melanoma differentiation associated gene-7 (mda-7)/Interleukin-24 (IL-24) (mda-7/IL-24) displays cancer-selective apoptosis induction when delivered via a replication-incompetent adenovirus (Ad.mda-7). In this study, the effect of Ad.mda-7 infection, either alone or in combination with gefitinib, was analyzed in a panel of NSCLC cell lines carrying wild-type EGFR (H 460 and H-2030) or mutant EGFR (H-1650 and H-1975). While H-2030 and H-1650 cells were sensitive, H-460 and H-1975 cells were resistance to growth inhibition by Ad.mda-7, which was reversed by the combination of Ad.mda-7 and gefitinib. This combination increased MDA-7/IL-24 and downstream effector double-stranded RNA activated protein kinase (PKR) protein expression, promoting apoptosis induction of NSCLC cells. Inhibition of PKR significantly inhibited apoptosis induction by Ad.mda-7 when administered alone but not when used in combination with gefitinib. The combination treatment also augmented inhibition of EGFR signaling. Our findings indicate that a combinatorial treatment with Ad.mda-7 and gefitinib may provide benefit in the treatment of NSCLC, especially in patients displaying resistance to clinically used EGFR inhibitors. PMID- 17111371 TI - Mitogen activated protein kinase-dependent activation of c-Jun and c-Fos is required for neuronal differentiation but not for growth and stress response in PC12 cells. AB - MAPK-dependent activation of AP-1 protein c-Jun is involved in PC12 cell differentiation and apoptosis. However, the role of other AP-1 proteins and their connection to MAPKs during growth, differentiation and apoptosis has remained elusive. Here we studied the activation of AP-1 proteins in response to ERK, JNK, and p38 signaling upon NGF, EGF and anisomycin exposures. All treatments caused different kinetics and strength of MAPK and AP-1 activities. NGF induced persistent ERK and AP-1 activities, whereas upon EGF and anisomycin exposures, their activities were only weakly and transiently induced. The sustained AP-1 activity was associated with concomitant c-Fos and c-Jun expression and phoshorylation, which were JNK and ERK dependent. While inhibition of the ERK, JNK, and p38 activities partially prevented AP-1 activity and suppressed differentiation, none of them was required for anisomycin-induced apoptosis. The importance of c-Fos and c-Jun as mediators of differentiation was demonstrated by the findings that the corresponding siRNAs suppressed NGF-induced neurite outgrowth. However, the capacity of c-Fos to promote differentiation required cooperation with Jun proteins. In contrast, Fra-2 expression was not required for the differentiation response. Together, the results show that sustained c-Jun and c-Fos activities mediate MAPK signaling and are essential for differentiation of PC12 cells. PMID- 17111372 TI - Retinal organization in the retinal degeneration 10 (rd10) mutant mouse: a morphological and ERG study. AB - Retinal degeneration 10 (rd10) mice are a model of autosomal recessive retinitis pigmentosa (RP), identified by Chang et al. in 2002 (Vision Res. 42:517-525). These mice carry a spontaneous mutation of the rod-phosphodiesterase (PDE) gene, leading to a rod degeneration that starts around P18. Later, cones are also lost. Because photoreceptor degeneration does not overlap with retinal development, and light responses can be recorded for about a month after birth, rd10 mice mimic typical human RP more closely than the well-known rd1 mutants. The aim of this study is to provide a comprehensive analysis of the morphology and function of the rd10 mouse retina during the period of maximum photoreceptor degeneration, thus contributing useful data for exploiting this novel model to study RP. We analyzed the morphology and survival of retinal cells in rd10 mice of various ages with quantitative immunocytochemistry and confocal microscopy; we also studied retinal function with the electroretinogram (ERG), recorded between P18 and P30. We found that photoreceptor death (peaking around P25) is accompanied and followed by dendritic retraction in bipolar and horizontal cells, which eventually undergo secondary degeneration. ERG reveals alterations in the physiology of the inner retina as early as P18 (before any obvious morphological change of inner neurons) and yet consistently with a reduced band amplification by bipolar cells. Thus, changes in the rd10 retina are very similar to what was previously found in rd1 mutants. However, an overall slower decay of retinal structure and function predicts that rd10 mice might become excellent models for rescue approaches. PMID- 17111373 TI - Rod bipolar cells and horizontal cells form displaced synaptic contacts with rods in the outer nuclear layer of the nob2 retina. AB - The nob2 mouse carries a null mutation in the Cacna1f gene, which encodes the pore-forming subunit of the L-type calcium channel, Ca(v)1.4. The loss of the electroretinogram b-wave in these mice suggests a severe reduction in transmission between photoreceptors and second-order neurons in the retina and supports a central role for the Ca(v)1.4 calcium channel at photoreceptor ribbon synapses, to which it has been localized. Here we show that the loss of Ca(v)1.4 leads to the aberrant outgrowth of rod bipolar cell dendrites and horizontal cell processes into the outer nuclear layer (ONL) of the nob2 retina and to the formation of ectopic synaptic contacts with rod photoreceptors in the ONL. Ectopic contacts are predominantly between rods and rod bipolar cells, with horizontal cell processes also present at some sites. Ectopic contacts contain apposed pre- and postsynaptic specializations, albeit with malformed synaptic ribbons. Cone photoreceptor terminals do not participate in ectopic contacts in the ONL. During retinal development, ectopic contacts appear in the days after eye opening, appearing progressively farther into the ONL at later postnatal stages. Ectopic contacts develop at the tips of rod bipolar cell dendrites and are less frequently associated with the tips of horizontal cell processes, consistent with the adult phenotype. The relative occurrence of pre- and postsynaptic markers in the ONL during development suggests a mechanism for the formation of ectopic synaptic contacts that is driven by the retraction of rod photoreceptor terminals and neurite outgrowth by rod bipolar cell dendrites. PMID- 17111374 TI - Epidermal growth factor receptor in adult retinal neurons of rat, mouse, and human. AB - During development, the epidermal growth factor receptor (EGFR) regulates proliferation and differentiation of many types of cells, including precursors of neurons and glia. In the adult, EGFR continues to drive the growth and differentiation of epithelial cells but is absent from glia in the CNS. However, the localization and functions of EGFR in adult neurons are not well defined. By using immunohistochemistry and Western blotting, we have identified EGFR and its ligands in adult retinal ganglion cells in the normal rat, mouse, and human retina. EGFR and its ligands were also present in certain other adult retinal neurons, for example, horizontal cells and amacrine cells, and had different distribution patterns among these species. In addition, we found that EGFR was expressed in the rat retinal ganglion cell line RGC-5. One of the EGFR ligands, EGF, caused a cell shape change and increased neurofilament phosphorylation in RGC-5 cells. The expression of EGFR in postmitotic, terminally differentiated adult retinal neurons suggests that EGFR has pleiotropic functions. In addition to the conventional mitogenic role in adult epithelial cells, EGFR must serve a different, nonmitogenic function in adult neurons. Our work localizes EGFR and its ligands in the adult retinas of several species as a step toward investigating the nonmitogenic functions of EGFR in adult neurons. PMID- 17111375 TI - Spatial relationship between synapse loss and beta-amyloid deposition in Tg2576 mice. AB - Although there is evidence that beta-amyloid impairs synaptic function, the relationship between beta-amyloid and synapse loss is not well understood. In this study we assessed synapse density within the hippocampus and the entorhinal cortex of Tg2576 mice at 6-18 months of age using stereological methods at both the light and electron microscope levels. Under light microscopy we failed to find overall decreases in the density of synaptophysin-positive boutons in any brain areas selected, but bouton density was significantly decreased within 200 mum of compact beta-amyloid plaques in the outer molecular layer of the dentate gyrus and Layers II and III of the entorhinal cortex at 15-18 months of age in Tg 2576 mice. Under electron microscopy, we found overall decreases in synapse density in the outer molecular layer of the dentate gyrus at both 6-9 and 15-18 months of age, and in Layers II and III of the entorhinal cortex at 15-18 months of age in Tg 2576 mice. However, we did not find overall changes in synapse density in the stratum radiatum of the CA1 subfield. Furthermore, in the two former brain areas we found a correlation between lower synapse density and greater proximity to beta-amyloid plaques. These results provide the first quantitative morphological evidence at the ultrastructure level of a spatial relationship between beta-amyloid plaques and synapse loss within the hippocampus and the entorhinal cortex of Tg2576 mice. PMID- 17111376 TI - Parallel thalamocortical pathways for echolocation and passive sound localization in a gleaning bat, Antrozous pallidus. AB - We present evidence for parallel auditory thalamocortical pathways that serve two different behaviors. The pallid bat listens for prey-generated noise (5-35 kHz) to localize prey, while reserving echolocation [downward frequency-modulated (FM) sweeps, 60-30 kHz] for obstacle avoidance. Its auditory cortex contains a tonotopic map representing frequencies from 6 to 70 kHz. The high-frequency (BF > 30 kHz) representation is dominated by FM sweep-selective neurons, whereas most neurons tuned to lower frequencies prefer noise. Retrograde tracer injections into these physiologically distinct cortical regions revealed that the high frequency region receives input from the suprageniculate (SG) nucleus, but not the ventral division of the medial geniculate body (MGBv), in all experiments (n = 9). In contrast, the low-frequency region receives tonotopically organized input from the MGBv in all experiments (n = 16). Labeling in the SG was observed in only two of these experiments. Both cortical regions also receive sparse inputs from medial (MGBm) and parts of the dorsal division (MGBd) outside the SG. These results show that the low- and high-frequency regions of a single tonotopic map receive dominant inputs from different thalamic divisions. Within the low frequency region, most neurons are binaurally inhibited, and an orderly map of interaural intensity difference (IID) sensitivity is present. We show that the input to the IID map arises from topographically organized projections from the MGBv. As observed in other species, a frequency-dependent organization is observed in the lateromedial direction in the MGBv. These data demonstrate that MGBv-to-auditory cortex connections are organized with respect to both frequency and binaural selectivity. PMID- 17111377 TI - Polarized distribution of ion channels within microdomains of the axon initial segment. AB - Voltage-gated sodium (Na(v)) channels accumulate at the axon initial segment (IS), where their high density supports spike initiation. Maintenance of this high density of Na(v) channels involves a macromolecular complex that includes the cytoskeletal linker protein ankyrin-G, the only protein known to bind Na(v) channels and localize them at the IS. We found previously that Na(v)1.6 is the predominant Na(v) channel isoform at IS of adult rodent retinal ganglion cells. However, here we report that Na(v)1.6 immunostaining is consistently reduced or absent in short regions of the IS proximal to the soma, although both ankyrin-G and pan-Na(v) antibodies stain this region. We show that this proximal IS subregion is a unique axonal microdomain, containing an accumulation of Na(v)1.1 channels that are spatially segregated from the Na(v)1.6 channels of the distal IS. Additionally, we find that axonal K(v)1.2 potassium channels are present within the distal IS, but are also excluded from the Na(v)1.1-enriched proximal IS microdomain. Because ankyrin-G was prominent in both proximal and distal subcompartments of the IS, where it colocalized with either Na(v)1.1 or Na(v)1.6, respectively, mechanisms other than association with ankyrin-G must mediate differential targeting of Na(v) channel subtypes to achieve the spatial precision observed within the IS. This precise arrangement of ion channels within the axon initial segment is likely an important determinant of the firing properties of ganglion cells and other mammalian neurons. PMID- 17111378 TI - Strategic expression of ion transport peptide gene products in central and peripheral neurons of insects. AB - Structurally related ion transport peptides (ITP) and crustacean hyperglycemic hormones (CHH) are increasingly implicated in diverse metabolic and developmental functions in arthropods. We identified a conserved ITP gene encoding two peptides by alternative splicing in Manduca sexta, Bombyx mori, and Aedes aegypti: A C terminally amidated ITP and a C-terminally unblocked ITP-like peptide (ITPL), which share common N-terminal sequences but have divergent C-termini. In the moth M. sexta, these peptides are expressed in two, regionally distinct neuronal populations in the central and peripheral nervous systems (CNS, PNS). MasITP expression is confined to the brain in five pairs of lateral neurosecretory cells (type Ia(2)) projecting ipsilateral axons into the retrocerebral complex and three to five pairs of adjacent small neurons that arborize extensively within the brain. Expression of MasITPL is comparatively weak in the brain but strong in the ventral ganglia and the PNS, where MasITP is absent. MasITPL occurs in bilaterally paired neurons of all thoracic and abdominal ganglia. In the PNS, MasITPL is coexpressed with crustacean cardioactive peptide in type II link nerve neurons (L1) of abdominal segments 2-7, which project axons into neurohemal transverse nerves. During metamorphosis, additional expression of MasITPL is observed in two pairs of small lateral neurons in the brain and one pair of ventromedial neurons in each of AG2-6. A similar pattern of differential ITP and ITPL expression was observed in the CNS and PNS of B. mori and Schistocerca americana. These distinctive cellular expression patterns suggest that ITP and ITPL have evolved specialized physiological functions in arthropods. PMID- 17111379 TI - The folate metabolic enzyme ALDH1L1 is restricted to the midline of the early CNS, suggesting a role in human neural tube defects. AB - Folate supplementation prevents up to 70% of human neural tube defects (NTDs), although the precise cellular and metabolic sites of action remain undefined. One possibility is that folate modulates the function of metabolic enzymes expressed in cellular populations involved in neural tube closure. Here we show that the folate metabolic enzyme ALDH1L1 is cell-specifically expressed in PAX3-negative radial glia at the midline of the neural tube during early murine embryogenesis. Midline restriction is not a general property of this branch of folate metabolism, as MTHFD1 displays broad and apparently ubiquitous expression throughout the neural tube. Consistent with previous work showing antiproliferative effects in vitro, ALDH1L1 upregulation during central nervous system (CNS) development correlates with reduced proliferation and most midline ALDH1L1(+) cells are quiescent. These data provide the first evidence for localized differences in folate metabolism within the early neural tube and suggest that folate might modulate proliferation via effects on midline Aldh1l1(+) cells. To begin addressing its role in neurulation, we analyzed a microdeletion mouse strain lacking Aldh1l1 and observed neither increased failure of neural tube closure nor detectable proliferation defects. Although these results indicate that loss-of-function Aldh1l1 mutations do not impair these processes in mice, the specific midline expression of ALDH1L1 and its ability to dominantly suppress proliferation in a folate responsive manner may suggest that mutations contributing to disease are gain-of-function, rather than loss-of function. Moreover, a role for loss-of-function mutations in human NTDs remains possible, as Mthfr null mice do not develop NTDs even though MTHFR mutations increase human NTD risk. PMID- 17111380 TI - Immunohistochemical localization of CNTFRalpha in adult mouse retina and optic nerve following intraorbital nerve crush: evidence for the axonal loss of a trophic factor receptor after injury. AB - Ciliary neurotrophic factor (CNTF) is important for the survival and outgrowth of retinal ganglion cells (RGCs) in vitro. However, in vivo adult RGCs fail to regenerate and subsequently die following axotomy, even though there are high levels of CNTF in the optic nerve. To address this discrepancy, we used immunohistochemistry to analyze the expression of CNTF receptor alpha (CNTFRalpha) in mouse retina and optic nerve following intraorbital nerve crush. In normal mice, RGC perikarya and axons were intensely labeled for CNTFRalpha. At 24 hours after crush, the immunoreactivity normally seen on axons in the nerve was lost near the lesion. This loss radiated from the crush site with time. At 2 days postlesion, labeled axons were not detected in the proximal nerve, and at 2 weeks were barely detectable in the retina. In the distal nerve, loss of axonal staining progressed to the optic chiasm by 7 days and remained undetectable at 2 weeks. Interfascicular glia in the normal optic nerve were faintly labeled, but by 24 hours after crush they became intensely labeled near the lesion. Double labeling showed these to be both astrocytes and oligodendrocytes. At 7 days postlesion, darkly labeled glia were seen throughout the optic nerve, but at 14 days labeling returned to normal. It is suggested that the loss of CNTFRalpha from axons renders RGCs unresponsive to CNTF, thereby contributing to regenerative failure and death, while its appearance on glia may promote glial scarring. PMID- 17111381 TI - Connections of functional areas in the mustached bat's auditory cortex with the auditory thalamus. AB - The auditory thalamus is the major target of the inferior colliculus and connects in turn with the auditory cortex. In the mustached bat, biosonar information is represented according to frequency in the central nucleus of the inferior colliculus (ICc) but according to response type in the cortex. In addition, the cortex has multiple areas with neurons of similar response type compared to the single tonotopic representation in the ICc. To investigate whether these transformations occur at the level of the thalamus, we injected anatomical tracers into physiologically defined locations in the mustached bat's auditory cortex. Injections in areas used for target ranging labeled contiguous regions of the auditory thalamus rather than separate patches corresponding to regions that respond to the different harmonic frequencies used for ranging. Injections in the two largest ranging areas produced labeling in separate locations. These results indicate that the thalamus is organized according to response type rather than frequency and that multiple mappings of response types exist. Injections in areas used for target detection labeled thalamic regions that were largely separate from those that interconnect with ranging areas. However, injections in an area used for determining target velocity overlapped with the areas connected to ranging areas and areas involved in target detection. Thus, separation by functional type and multiplication of areas with similar response type occurs by the thalamic level, but connections with the cortex segregate the functional types more completely than occurs in the thalamus. PMID- 17111382 TI - Identification of differentially expressed microRNAs by microarray: a possible role for microRNA genes in pituitary adenomas. AB - MicroRNAs (miRNAs) are small non-coding RNAs that control gene expression by targeting mRNA. It has been demonstrated that miRNA expression is altered in many human cancers, suggesting that they may play a role in human neoplasia. To determine whether miRNA expression is altered in pituitary adenomas, we analyzed the entire miRNAome in 32 pituitary adenomas and in 6 normal pituitary samples by microarray and by Real-Time PCR. Here, we show that 30 miRNAs are differentially expressed between normal pituitary and pituitary adenomas. Moreover, 24 miRNAs were identified as a predictive signature of pituitary adenoma and 29 miRNAs were able to predict pituitary adenoma histotype. miRNA expression could differentiate micro- from macro-adenomas and treated from non-treated patient samples. Several of the identified miRNAs are involved in cell proliferation and apoptosis, suggesting that their deregulated expression may be involved in pituitary tumorigenesis. Predictive miRNAs could be potentially useful diagnostic markers, improving the classification of pituitary adenomas. PMID- 17111383 TI - Lipid raft-dependent endocytosis of metallothionein in HepG2 cells. AB - Human hepatocellular carcinoma (HepG2) cells take up metallothionein (MT) by endocytosis. MT co-localizes with albumin but not with transferrin, indicating uptake via a non-classical pathway rather than via clathrin-mediated endocytosis. A lipid raft-dependent uptake is indicated by pravastatin inhibition of cholesterol synthesis and methyl-beta-cyclodextrin inhibition of cholesterol translocation to the plasma membrane, reducing MT uptake by 29% and 69%, respectively. Subcellular fractionation after MT uptake reveals significant amounts of MT in vesicular fractions including lysosomes but virtually no MT in the cytosol. Metals bound to MT are released into the cytosol, however. The findings define a pathway for cellular metal acquisition. Together with results from other studies demonstrating secretion of MT from different cells and the presence of MT in extracellular fluids, the results suggest a function of MT in intercellular communication. PMID- 17111384 TI - Heterogeneity of aldehyde dehydrogenase expression in lung cancer cell lines is revealed by Aldefluor flow cytometry-based assay. AB - BACKGROUND: We have been interested in studying the roles of two aldehyde dehydrogenases in the biology of lung cancer. In this study, we seek to apply Aldefluor flow cytometry-based assay for the measurement of aldehyde dehydrogenase (ALDH) activity in lung cancer cell lines, which may become a new tool that will facilitate our continued research in this field. EXPERIMENTAL DESIGN: Several established lung cancer cell lines were used, including A549 cell line expressing siRNA against aldehyde dehydrogenase class-1A1 (ALDH1A1). Western blot analysis, spectrophotometry assay, and Aldefluor staining were used to measure protein or enzyme activity in these cell lines. For the purpose of measurement of ALDH activity by Aldefluor in cells with known high ALDH levels, cells were mixed 1:10 with immortalized lung epithelial cell line (Beas-2B), which is known to lack ALDH activity. To delineate dead cells, double staining using Aldefluor and propidium iodide (PI) was done. Double staining was also used to detect changes in ALDH activity in two different cell lines after treatment with 4-hydroperoxycyclophosphamide (4-HC). RESULTS: Our results show a very good correlation between Aldefluor, Western blot, and spectrophotometry assays. Mixing experiments with Beas-2B cells allowed accurate assessment of ALDH activity in A549 cells at baseline and after siRNA expression, thus establishing an approach that facilitates the measurement of very high ALDH using the Aldefluor assay. Aldefluor staining was able to detect heterogeneity in ALDH expression among as well as within the same cell lines and better assess viability after 4-HC treatment when combined with PI. CONCLUSIONS: Aldefluor assay can be adapted successfully to measure ALDH activity in lung cancer cells and may have the advantage of providing real time changes in ALDH activity in viable cells treated with siRNA or chemotherapy. PMID- 17111385 TI - The apoptosis of blood polymorphonuclear leukocytes in sickle cell disease. AB - BACKGROUND: The apoptosis of human polymorphonuclear leukocytes (PMNs) in patients with sickle cell disease (SCD) is not well understood. The goal of this study was to examine the apoptosis of PMNs in patients with SCD and in controls. METHODS: Flow cytometric quantitation of PMN apoptosis was performed in 17 patients during and after sickle cell vasoocclusive crisis and in 17 healthy volunteers. Plasma nitric oxide concentrations were also measured in patients with SCD. RESULTS: The mean of annexin-V and annexin-V/PI staining (early and late apoptotic cells) increased to a greater degree in patients with SCD than in healthy controls for patients with SCD during and after vasoocclusive crisis. The mean of PI staining showing dead cells was higher only in patients after SCD crisis than in healthy controls. In the SCD groups during and after vasoocclusive crisis, there was no difference between PMN apoptosis levels. Furthermore, plasma nitric oxide concentrations were not correlated with PMN apoptosis. CONCLUSIONS: There was an evidence that the alteration of blood PMN apoptosis could contribute to the pathogenetic mechanisms of vasoocclusion in patients with SCD. This can be attributed to the effects of numerous inflammatory mediators rather than simply the effects of nitric oxide. PMID- 17111386 TI - Flow-assisted quantification of in vitro activated basophils in the diagnosis of wasp venom allergy and follow-up of wasp venom immunotherapy. AB - BACKGROUND: Correct identification of the culprit venom is a prerequisite for specific venom immunotherapy (VIT). Despite the efficacy of VIT, issues as how to monitor treatment and when to discontinue maintenance therapy remain to be established. METHODS: To evaluate diagnostic performances of the basophil activation test (BAT) in wasp venom allergy, 80 patients with a definite history of wasp venom anaphylaxis (systemic reactors) and 14 wasp-stung asymptomatic controls (stung controls) were enrolled. Venom-induced basophil activation was analyzed flow cytometrically by double-labeling with anti-IgE and anti-CD63. Results were compared to wasp IgE levels and results of a venom skin test (VST). To establish whether the BAT constitutes a candidate marker to monitor VIT, the BAT was repeated in 22 patients on the 5th day of a build-up course and after 6 months of maintenance VIT. Whether the BAT could contribute in the decision of discontinuing VIT was assessed in a cross-sectional analysis in 30 patients receiving treatment for 3 years. RESULTS: Comparison between systemic reactors and stung controls revealed a sensitivity of 86.4% and specificity of 100% for venom IgE, and sensitivity of 81.8% for VST, respectively. In contrast to stung controls, patients demonstrated dose-dependent venom-induced basophil activation. The BAT attained a sensitivity of 83.8% and specificity of 100%. At the end of the build-up course, no effect of VIT on the BAT was demonstrable. When the BAT was repeated after 6 months of treatment, submaximal stimulation of the cells demonstrated a significant decreased CD63 expression (P < 0.04). Patients having VIT for 3 years also demonstrated significantly lower venom-induced CD63 expression (P < 0.001). After 3 years, 60% of the patients had a negative BAT for submaximal stimulation of the cells whereas only 17.9% of the patients had negativation of wasp IgE. CONCLUSIONS: The BAT is a reliable instrument for the diagnosis of wasp venom anaphylaxis and might constitute an instrument to monitor wasp VIT. PMID- 17111387 TI - Are the vulnerability effects of personality and psychosocial functioning on depression accounted for by subthreshold symptoms? AB - Previous studies could not evaluate adequately the extent to which deviant levels of personality measures and psychosocial functioning found before and after a major depressive episode (MDE) should be attributed to subthreshold depressive symptoms. Our aim is to investigate whether pre- and post-MDE personality alterations and psychosocial disability truly reflect vulnerability, or whether they can be accounted for by the presence of subthreshold depressive symptoms. Data were derived from the Netherlands Mental Health and Incidence Study, a prospective general population study with three waves. Psychopathology was measured with the Composite International Diagnostic Interview (CIDI). Course of depressive symptoms was assessed with the Life Chart Instrument in a cohort of 195 respondents with a new or recurrent MDE between waves 2 and 3. Personality and psychosocial functioning were assessed with, respectively, four and two different measures. Alterations in measurements of personality and psychosocial functioning were present before onset and after remission of an MDE. Most pre- and postonset alterations occurred in the presence of subthreshold depressive symptoms. But even without these subthreshold symptoms, some alterations in measurements of personality and psychosocial functioning were found before and after an MDE. Depressive complaints between waves 2 and 3 were retrospectively assessed, and only a limited set of brief questionnaires was used to assess personality styles and psychosocial functioning. It is unlikely that the pre- and post-MDE alterations in personality and psychosocial functioning observed in earlier studies are entirely due to subthreshold depressive symptoms. This suggests that a depressive episode is interwoven in a long-standing and enduring pattern of mild personality deviance and limitations in psychosocial functioning. PMID- 17111388 TI - Aripiprazole augmentation for treatment of patients with inadequate antidepressants response. AB - This study evaluated whether or not augmentation with aripiprzazole is beneficial and tolerable to patients with an inadequate response to antidepressants (ADs). Thirteen patients with nonpsychotic major depression, who had failed to respond to an adequate trial of at least one AD, were prescribed aripiprazole (dose, 5-30 mg) for 8 weeks. The dose of their preexisting ADs was not changed. The treatment response was defined as the mean changes in the scores of the Hamilton Depression Rating Scale (HAM-D) from the baseline to the end of treatment. Eleven (84.6%) patients returned for at least one follow-up visit, and 7 (53.8%) patients completed the study. The HAM-D and Clinical Global Impression-Severity (CGI-S) scores decreased significantly from the baseline to the end of treatment by 53.8% and 56.0%, respectively (Z = -2.937, P =.003; Z = -2.961, P =.003). Seven (63.6%) patients showed a > or = 50% reduction in the HAM-D score at the end of treatment. Three (27.3%) patients met the remission criteria at the end of treatment. There were no serious side effects. Despite the high dropout rate in this open study, aripiprazole appears to be reasonably effective and tolerated as an augmentation strategy in conjunction with conventional ADs treatment in patients with an inadequate AD response. These results highlight the potential benefits of aripiprazole for these patients. However, adequately powered, randomized, controlled trials are needed to confirm these results. PMID- 17111389 TI - Management of patients with resectable stage IIIA (N2) NSCLC. PMID- 17111390 TI - Lessons from the old masters: pragmatism or purity, FDG PET SUV, serum glucose and prediction of nodal status in non-small cell lung cancer. PMID- 17111391 TI - Combined ductal lavage and ductoscopy: what is the future for the intraductal approach? PMID- 17111392 TI - Significance of elevated preoperative alpha-fetoprotein in postchemotherapy residual tumor resection for the disseminated germ cell tumors. AB - BACKGROUND AND OBJECTIVES: The purpose of the study is to determine the significance of elevated serum alpha-fetoprotein (AFP) in the setting prior to residual tumor resection (RTR) following chemotherapy for metastatic germ cell tumor in terms of the prediction of histology of the specimen and postoperative survival. METHODS: We conducted a retrospective review of 68 patients undergoing RTR for metastatic nonseminomatous germ cell tumor or extragonadal germ cell tumor after at least a first-line chemotherapy. Pretreatment and postchemotherapy serum markers were evaluated in association with other clinical findings including results of pathological examination of RTR specimen and surgical outcome. RESULTS: Of the 68 study patients, 54 (79%) and 45 (66%) had positive AFP and beta-human chorionic gonadotropin (beta-HCG) in pretreatment settings. Rates of presence of residual malignant cell in RTR specimen were similar between patients with normal AFP (7/28 or 25%) and with mildly elevated (10-30 ng/ml) AFP (3/11 or 27%). In 26 patients who had residual viable malignancy in RTR specimen, patients with preoperative positive AFP had significantly better survival (P = 0.02) compared to those with preoperative positive beta-HCG. CONCLUSIONS: Sole and mild elevation of AFP is not always associated with postoperative poor prognosis. It should be carefully considered individually whether a mild elevation of AFP after chemotherapy represents residual malignancy or benign pathogenesis. PMID- 17111393 TI - Radiotherapy for verrucous carcinoma of the oral cavity. PMID- 17111394 TI - Vascular endothelial growth factor polymorphisms in gastric cancer development, prognosis, and survival. AB - BACKGROUND: Angiogenesis plays an important role in growth, progression, and metastasis of tumors. Vascular endothelial growth factor (VEGF) overexpression has been associated with advanced stage and poor survival in several cancers. METHODS: The association of functional single nucleotide polymorphisms (SNPs) of VEGF gene with gastric cancer development, prognosis, and survival in a case control study of 100 gastric cancer Greek patients was evaluated. The genotyping was performed by PCR-RFLP analysis. RESULTS: Our results revealed a marginally significant association of the 634CC genotype (P = 0.042) with increased risk for gastric cancer development. None of the rest of the examined polymorphisms or haplotypes conferred any gastric cancer susceptibility. A strong association between the -2578AA (P = 0.025), -634CC (P = 0.013), +936CT (P = 0.028), +936TT (P = 0.0001) genotypes and a larger tumor size was observed, while the 2578AA and -634CC genotypes were strongly correlated to poor differentiation (P = 0.01) and advanced stage of disease (P = 0.039), respectively. In addition, our results indicated that metastatic disease frequency was more accentuated among the +936TT carriers (P = 0.0035). Interestingly, carrying the -634CC genotype was associated with decreased overall survival rates (46.67%). CONCLUSIONS: The present study suggests that VEGF polymorphisms may contribute to gastric tumor characteristics; these observations, however, requiring further confirmation in a larger multi ethnic study. PMID- 17111395 TI - Comprehensive analysis of chemical bonding in boron clusters. AB - We present a comprehensive analysis of chemical bonding in pure boron clusters. It is now established in joint experimental and theoretical studies that pure boron clusters are planar or quasi-planar at least up to twenty atoms. Their planarity or quasi-planarity was usually discussed in terms of pi-delocalization or pi-aromaticity. In the current article, we demonstrated that one cannot ignore sigma-electrons and that the presence of two-center two-electron (2c--2e) peripheral B--B bonds together with the globally delocalized sigma-electrons must be taken into consideration when the shape of pure boron cluster is discussed. The global aromaticity (or global antiaromaticity) can be assigned on the basis of the 4n+2 (or 4n) electron counting rule for either pi- or sigma-electrons in the planar structures. We showed that pure boron clusters could have double (sigma- and pi-) aromaticity (B3-, B4, B5+, B6(2+), B7+, B7-, B8, B(8)2-, B9-, B10, B11+, B12, and B13+), double (sigma- and pi-) antiaromaticity (B6(2-), B15), or conflicting aromaticity (B5-,sigma-antiaromatic and pi-aromatic and B14, sigma aromatic and pi-antiaromatic). Appropriate geometric fit is also an essential factor, which determines the shape of the most stable structures. In all the boron clusters considered here, the peripheral atoms form planar cycles. Peripheral 2c--2e B--B bonds are built up from s to p hybrid atomic orbitals and this enforces the planarity of the cycle. If the given number of central atoms (1, 2, 3, or 4) can perfectly fit the central cavity then the overall structure is planar. Otherwise, central atoms come out of the plane of the cycle and the overall structure is quasi-planar. PMID- 17111396 TI - Effects of ethylene glycol on the torsion elastic constant and hydrodynamic radius of p30delta DNA. AB - Upon increasing the concentration of ethylene glycol (EG) at 37 degrees C, the twist energy parameter, E(T), which governs the supercoiling free energy, was recently found to undergo a decreasing (or reverse) sigmoidal transition with a midpoint near 20 w/v % EG. In this study, the effects of adding 20 w/v % EG on the torsion elastic constant (alpha) of linear p30delta DNA and on the hydrodynamic radius (R(H)) of a synthetic 24 bp duplex DNA were examined at both 40 and 20 degrees C. The time-resolved fluorescence intensity and fluorescence polarization anisotropy (FPA) of intercalated ethidium were measured in order to assess the effects of 20 w/v % EG on: (1) alpha; (2) R(H); (3) the lifetimes of intercalated and non-intercalated dye; (4) the amplitude of dye wobble in its binding site; and (5) the binding constant for intercalation. The effects of 20 w/v % EG on the circular dichroism (CD) spectrum of the DNA and on the emission spectrum of the free dye were also measured. At 40 degrees C, addition of 20 w/v % EG caused a substantial (1.27- to 1.35-fold) increase in alpha, a significant change in the CD spectrum, and a very small, marginally significant increase in R(H), but little or no change in the amplitude of dye wobble in its binding site or the lifetime of intercalated dye. Together with previously reported measurements of E(T), these results imply that the bending elastic constant of DNA is significantly decreased by 20 w/v % EG at 40 degrees C. At 20 degrees C, addition of 20 w/v % EG caused a marginally significant decrease in alpha and very little change in any other measured properties. Also at 20 degrees C, addition of 30 w/v % betaine caused a marginally significant increase in alpha and significant but modest change in the CD spectrum, but very little change in any other properties. PMID- 17111397 TI - Future of biomedical sciences: single molecule microscopy. AB - The behavior of single molecule defines whether a cell lives, dies, or responds to a specific drug treatment. Single molecule microscopies have begun to reveal the number, location, and functionalities of molecules outside and inside living cells. This issue of Biopolymers presents a first set of reviews that aim to highlight the accomplishments and future prospects of single molecule microscopies. PMID- 17111398 TI - Fetal programming of temperamental negative affectivity among children born healthy at term. AB - The fetal programming hypothesis suggests that an adverse in utero environment, reflected in small body size at birth, has life-long effects on different physiological systems that may affect both health and behavior. We explored whether fetal growth was associated with biologically based temperamental outcomes (negative affectivity scales, the CBQ) among 5(1/2)-year-old children (n = 416) born healthy at term (gestational weeks 37-42). In line with the hypotheses, small body size at birth (thinness measured by ponderal index, kg/m(3)) was related to increased negative affectivity and its subscales: anger-, discomfort-, and sadness-proneness in childhood. Longer length at birth was predictive of higher levels of child anger- and sadness-proneness. Length of gestation moderated the associations of weight and length at birth with negative affectivity. The results suggest that the biological basis of temperament may be subjected to antenatal environmental influences, and that the mechanisms, proposed to be related to fetal glucocorticoid environment, may operate even within the normal range of term birth. PMID- 17111399 TI - Judgements of facial and vocal signs of emotion in infants with Down syndrome. AB - We address how adults perceive facial and vocal signs of emotions in infants with and without Down syndrome. A set of naturalistic data from infants with trisomy 21 and typically developing infants (joy expression of young infants, 3.8-4.4 months, and anger and neutral expressions of older infants, 6.8-12.8 months) was rated by adult judges categorically or dimensionally. Facial signs alone, vocal signs alone, and both facial and vocal signs were presented for each expression. Raters were university students who did not have regular contact with infants, nor with people with mental retardation. Young infants' joy expressions were correctly recognized more frequently for typically developing infants than for infants with Down syndrome and, specifically, joy vocalizations in infants with Down syndrome were not identified. Facial signs were also more communicative than vocal signs in the case of older infants' anger and neutral expressions. These results are relevant to the way infant emotion is perceived by others, and may be particularly useful in facilitating interaction between adults and infants with Down syndrome. PMID- 17111400 TI - Brief exposure to the biological mother "potentiates" the isolation behavior of precocial Guinea pig pups. AB - When isolated rat pups are briefly reunited with a lactating female, her subsequent removal leads to a dramatic increase in the emission of ultrasonic vocalizations, but not other behaviors. Whether this socially induced augmentation of isolation behavior (i.e., "potentiation") is characteristic only of altricial rodents is not known. Therefore, we examined precocial guinea pig pups in a potentiation paradigm. Ten-day-old guinea pigs were isolated in a test cage for 10 min, at which time they were then placed into a second cage for 5 min that either contained a companion or, for controls, was empty. Pups were then isolated again in the test cage for a second 10-min period. Control pups showed a significant reduction in vocalizing and locomotor activity from the first to second isolation period. Exposure to the biological mother prevented the decline in both behaviors (Experiment 1), whereas exposure to a familiar littermate (Experiment 2) had no effect, and exposure to an unfamiliar lactating female (Experiment 3) had only a minimal effect on locomotor activity. The results show that potentiation of isolation behaviors is not limited to altricial rodents, and suggest that specific characteristics of the effect (i.e., its magnitude, the specific behaviors affected, and the selectivity of the response to particular social partners) varies with the abilities and requirements of the young, as well as the behavioral ecology of the species in question. PMID- 17111401 TI - Short-term plasticity in children's speech motor systems. AB - Speech production is a highly skilled behavior that requires rapid and coordinated movements of the orofacial articulators. Previous studies of speech development have shown that children have more variable articulatory movements compared to adults, and cross-sectional studies have revealed that a gradual transition to more stable movement patterns occurs with age. The focus of the present investigation is on the potential role of short-term changes in speech motor performance related to practice. Thus we developed a paradigm to examine the influences of phonological complexity and practice on children (9 and 10-year olds) and adults' production of novel nonwords. Using two indices that reflect the degree of trial-to-trial consistency of articulatory movements, we analyzed the first and last five productions of the novel nonwords. Both children and adults accurately produced the novel nonwords; however, children showed a practice effect; their last five trials were more consistently produced than their first five trials. Adults did not show this practice effect. This study provides new evidence that children show short-term changes in their speech coordinative patterns with practice. In addition, the present findings support the contribution of neuromotor noise or background, inherent variability to speech motor development. PMID- 17111402 TI - Influence of various early human-foal interferences on subsequent human-foal relationship. AB - Whereas the way animals perceive human contact has been particularly examined in pet animals, a small amount of investigations has been done in domestic ungulates. It was nevertheless assumed that, as pet animals, non-aggressive forms of tactile contact were as well rewarding or positive for these species, even though the features of intraspecific relationships in pet animals and domestic ungulates may be to some extent different. We test here the hypothesis that horses may not consider physical handling by humans as a positive event. When comparing different early human-foal interactions, we found that early exposure to a motionless human enhanced slightly foals reactions to humans whereas forced stroking or handling in early life did not improve later human-foal relation. Foals that were assisted during their first suckling (e.g., brought to the dam's teat) even tended to avoid human approach at 2 weeks, and physical contact at 1 month of age. We argue that interspecies differences may exist in how tactile stimulation is perceived. It may be important for the establishment of a bond that a young animal is active in the process and able, through its behavioral responses, to help define what is positive for it. This way of investigation may have important general implications in how we consider the development of social relations, both within and between species. PMID- 17111403 TI - Parity does not alter baseline or stimulated activity of the hypothalamus pituitary-adrenal axis in women. AB - Pregnancy is associated with considerable physiological adaptations, some of which long outlast the state of pregnancy. Although it is well documented that pregnancy produces alterations of the hypothalamus-pituitary-adrenal axis, the longer-term effects of pregnancy on this system have not been systematically examined in humans. Subjects in the present study were 159 nulliparous and 265 parous women. Data analysis revealed no impact of parity on baseline activity (salivary cortisol: response to awakening, F RGDSVVYGLR approximately RGDS > RGES. Using blocking monoclonal antibodies, cellular adhesion to the peptides was shown to be primarily alpha(v)-integrin-mediated. In contrast, the tau(50) value of the cells on fibronectin (Fn)-coated substrates of similar surface density was 6-7 times higher and involved both alpha(5)beta(1) and alpha(v)beta(3) integrins. Cellular spreading was enhanced on RGD peptides after 1 h when compared to RGE and unmodified substrates. However, no significant differences were observed between the different RGD peptides. Long-term function of MC3T3-E1 cells was also evaluated by measuring alkaline phosphatase (ALP) activity and mineral deposition. Among the four peptides, RGDSPK exhibited the highest level of ALP activity after 11 days and mineralization after 15 days and reached comparable levels as Fn substrates after 15 and 24 days, respectively. These findings collectively illustrate both the advantages and limitations of enhancing cellular adhesion and function by the design of RGD peptides. PMID- 17111409 TI - Children's memory for a mild stressor: the role of sympathetic activation and parasympathetic withdrawal. AB - Although numerous studies have examined the relations between stress and memory in children, few studies have investigated physiological responses as predictors of children's memory for stressful events. In this study, 4- to 8-year-olds completed laboratory challenges and experienced a fire-alarm incident while their sympathetic and parasympathetic reactions were monitored. Shortly afterward, children's memory of the alarm incident was tested. As children's age and family income increased, memory performance improved. High sympathetic activation during the laboratory challenges was associated with enhanced memory. Also, a trend indicated that, among older children, greater general parasympathetic withdrawal was associated with poorer memory, but among younger children, parasympathetic withdrawal was unrelated to memory. Findings highlight the need to measure both sympathetic and parasympathetic responses when evaluating children's memory for mild stressors and to include a wide age range so that developmental changes in the relations between stress and memory in childhood can be identified. PMID- 17111410 TI - In situ analysis of breast cancer progression in murine models using a macroscopic fluorescence imaging system. AB - BACKGROUND AND OBJECTIVE: The goal of this study was to use an inexpensive macroscopic imaging system to monitor tumor progression in mouse models in real time with minimal intervention. STUDY DESIGN/MATERIALS AND METHODS: Illumination is provided via a xenon arc lamp and a fiber optic probe which delivers white light or quasi-monochromatic excitation via specific bandpass filters. Fluorescence emission from SCID and nude mice following mammary fat pad injection of red fluorescence protein (RFP)-expressing human breast cancer cell lines was recorded and quantified using a single lens reflex (SLR) digital camera. RESULTS: This simple system enabled the verification of successful tumor take and temporal quantification of tumor progression in mouse models. CONCLUSION: The macroscopic fluorescence imaging system represents an inexpensive and portable tool to facilitate non-invasive in situ cancer detection with the potential to monitor fluorescent tumor formation and investigation of the efficacy of potential cancer therapeutics. PMID- 17111411 TI - Impaired maze performance in aged rats is accompanied by increased density of NMDA, 5-HT1A, and alpha-adrenoceptor binding in hippocampus. AB - Using quantitative receptor autoradiography, we assessed binding site densities and distribution patterns of glutamate, GABA(A), acetylcholine (ACh), and monoamine receptors in the hippocampus of 32-month-old Fischer 344/Brown Norway rats. Prior to autoradiography, the rats were divided into two groups according to their retention performance in a water maze reference memory task, which was assessed 1 week after 8 days of daily maze training. The animals of the inferior group showed less long-term retention of the hidden-platform task but did not differ from superior rats in their navigation performance during place training and cued trials. The decreased retention performance in the group of inferior learners was primarily accompanied by increased alpha(1)-adrenoceptors in all hippocampal subregions under inspection (CA1-CA4 and dentate gyrus), while elevated alpha(2)-adrenoceptor binding was observed in the CA1 region and DG. Furthermore, inferior learners had higher NMDA binding in the CA2 and CA4 and increased 5-HT(1A) binding sites in the CA2, CA3, and CA4 region. No significant differences between inferior and superior learners were evident with regard to AMPA, kainate, GABA(A), muscarinergic M(1), dopamine D(1), and 5-HT(2) binding densities in any hippocampal region analyzed. These results show that increased NMDA, 5-HT(1A), and alpha-adrenoceptor binding in the hippocampus is associated with a decline in spatial memory. The increased receptor binding observed in the group of old rats with inferior maze performance might be the result of neural adaptation triggered by age-related changes in synaptic connectivity and/or synaptic activity. PMID- 17111412 TI - Post-translational synaptic protein modification as substrate for long-lasting, remote memory: an initial test. AB - The current view of the molecular basis for information storage is that post translational modification (PTM) of brain proteins is important for the early stages of memory storage and that protein synthesis is necessary for long-lasting memory. This view has been challenged by the proposal that PTM of synaptic proteins is the critical instructive mechanism underlying both recent as well as long-lasting memories (Routtenberg and Rekart, 2005). As an initial test, a broad spectrum serine/threonine kinase inhibitor (H-7) was delivered bilaterally to rat anterior cingulate cortex 1 h before a 3 week retention test of contextual fear conditioning. This significantly blocked 21-Day retention. In the second experiment evaluating extinction of a 21-Day remote memory, H-7 injected into mouse medial prefrontal cortex blocked fear extinction. As the H-7-induced impairment in 21-Day retention was indexed by a decrease in freezing, while the extinction blockade by no decrease in freezing, the results could not be ascribed to a direct effect of the drug on behavioral performance. This represents the first demonstration, to our knowledge, that PTM inhibition, here serine/threonine kinase activity, interferes with long-lasting memory, providing initial support for the PTM model. PMID- 17111413 TI - An experimental pathologic study of gingivectomy using dual-wavelength laser equipment with OPO. AB - BACKGROUND AND OBJECTIVES: This study was conducted to evaluate how soft tissues respond to treatment by a tunable laser with an optical parametric oscillating (OPO) mechanism capable of simultaneously emitting two wavelengths. MATERIALS AND METHODS: Marginal gingiva of dogs was incised by a prototype laser oscillator. The oscillator was set at two wavelengths known to effectively incise tissue and arrest hemorrhage with minimal invasiveness. Four laser irradiation conditions were set based on different combinations of the 1.67 and 2.94 microm wavelengths. The animals were sacrificed immediately after surgery, 7 days after surgery, and 28 days after surgery for histological examination. RESULTS: When irradiation at 1.67 and 2.94 microm wavelengths was simultaneously applied, the former conferred an observable hemostatic effect and the latter incised the tissue. Wound healing was similar to that in conventional methods and no serious inflammation was observed. CONCLUSION: Simultaneous irradiation at wavelengths of 1.67 and 2.94 microm can be an effective method in soft tissue surgery. PMID- 17111414 TI - Tissue factor and thrombomodulin expression on endothelial cell-seeded collagen modules for tissue engineering. AB - The creation of functional tissue engineering constructs to repair or replace diseased tissues requires a well-formed vasculature network within the construct and the endothelial cells lining that vascular bed must display a nonthrombogenic phenotype. A new approach to tissue engineering involves the assembly of smaller components (modules fabricated at the hundred micron scale) into larger constructs. The modules, collagen gel containing the particular tissue cell of interest, are covered with endothelial cells prior to assembly so that the interconnected channels that are formed are lined with endothelial cells, creating a mimic of a vascular network. Here, we confirmed (using confocal microscopy primarily) that the human umbilical vein endothelial cells, seeded on collagen gel modules without a second embedded cell and without flow, bore the molecular markers of low thrombogenicity. Two days, after seeding on the modules, endothelial cells displayed the typical cobblestone morphology, formed tight cell cell junctions and covered the whole module surface. Immunofluorescence staining showed that at both 2 days and 7 days after seeding, only a few cells expressed tissue factor while this number was dramatically increased after TNFalpha stimulation. On the other hand, thrombomodulin was expressed by the majority of seeded cells and expression was reduced after TNFalpha stimulation. PMID- 17111415 TI - Blue and red light combination LED phototherapy for acne vulgaris in patients with skin phototype IV. AB - BACKGROUND AND OBJECTIVES: Blue light is effective for acne treatment, inducing photodynamic destruction of Propionibacterium acnes (P. acnes). This study was designed to investigate the efficacy of combined blue and red light-emitting diode (LED) phototherapy for acne vulgaris. MATERIALS AND METHODS: Twenty-four patients with mild to moderately severe facial acne were treated with quasimonochromatic LED devices, alternating blue (415 nm) and red (633 nm) light. The treatment was performed twice a week for 4 weeks. Objective assays of the skin condition were carried out before and after treatment at each treatment session. Clinical assessments were conducted before treatment, after the 2nd, 4th, and 6th treatment sessions and at 2, 4, and 8 weeks after the final treatment by grading and lesion counting. RESULTS: The final mean percentage improvements in non-inflammatory and inflammatory lesions were 34.28% and 77.93%, respectively. Instrumental measurements indicated that the melanin levels significantly decreased after treatment. Brightened skin tone and improved skin texture were spontaneously reported by 14 patients. CONCLUSION: Blue and red light combination LED phototherapy is an effective, safe and non-painful treatment for mild to moderately severe acne vulgaris, particularly for papulopustular acne lesions. PMID- 17111416 TI - Laser-induced thermotherapy (LITT) elevates mRNA expression of connective tissue growth factor (CTGF) associated with reduced tumor growth of liver metastases compared to hepatic resection. AB - BACKGROUND AND OBJECTIVES: Proliferation and synthesis of hepatocellular tissue after tissue damage are promoted by specific growth factors such as hepatic tissue growth factor (HGF) and connective growth factor (CTGF). Laser-induced thermotherapy (LITT) for the treatment of liver metastases is deemed to be a parenchyma-saving procedure compared to hepatic resection. The aim of this study was to compare the impact of LITT and hepatic resection on intrahepatic residual tumor tissue and expression levels of mRNA HGF/CTGF within liver and tumor tissue. STUDY DESIGN/MATERIALS AND METHODS: Two independent adenocarcinomas (CC531) were implanted into 75 WAG rats, one in the right (untreated tumor) and one in the left liver lobe (treated tumor). The left lobe tumor was treated either by LITT or partial hepatectomy. The control tumor was submitted to in-situ hybridization of HGF and CTGF 24-96 hours and 14 days after intervention. RESULTS: Volumes of the untreated tumors prior to intervention were 38+/-8 mm(3) in group I (laser), 39 +/- 7 mm(3) in group II (resection), and 42 +/- 12 mm(3) in group III (control) and did not differ significantly (P > 0.05). Fourteen days after the intervention the mean tumor+/-SEM volume of untreated tumor in group I (laser) [223 +/- 36] was smaller than in group II (resection) [1233.28 +/- 181.52; P < 0.001], and in group III (control) [978.92 +/- 87.57; P < 0.003]. Forty-eight hours after the intervention intrahepatic mRNA expression level of HGF in group II (resection) was almost twofold higher than in group I (laser) [7.2 +/- 1.0 c/mf vs. 3.9 +/- 0.4 c/mf; P<0.01]. Fourteen days after the intervention intrahepatic mRNA expression level of CTGF in group I (laser) was higher than in group II (resection) [13.89 +/- 0.77 c/mf vs. 9.09 +/- 0.78 c/mf; P < 0.003]. CONCLUSIONS: LITT leads to a decrease of residual tumor growth in comparison to hepatic resection. Accelerated tumor growth after hepatic resection is associated with higher mRNA level of HGF and reduced tumor growth after LITT with higher mRNA level of CTGF. The increased CTGF-mediated regulation of ECM may cause reduced residual tumor growth after LITT. PMID- 17111417 TI - Tomographic visualization of cholinesterase. PMID- 17111418 TI - Successful peripheral blood stem cell harvest on a 5.5-kg infant. PMID- 17111419 TI - Severe citrate toxicity complicating volunteer apheresis platelet donation. AB - We report a case of severe citrate toxicity during volunteer donor apheresis platelet collection. The donor was a 40-year-old female, first-time apheresis platelet donor. Past medical history was remarkable for hypertension, hyperlipidemia, and depression. Reported medications included bumetanide, pravastatin, and paroxetine. Thirty minutes from the start of the procedure, the donor noted tingling around the mouth, hands, and feet. She then very rapidly developed acute onset of severe facial and extremity tetany. Empirical treatment with intravenous calcium gluconate was initiated, and muscle contractions slowly subsided over approximately 10 to 15 minutes. The events are consistent with a severe reaction to calcium chelation by sodium citrate anticoagulant resulting in symptomatic systemic hypocalcemia. Upon additional retrospective analysis, it was noted that bumetanide is a loop diuretic that may cause significant hypocalcemia. We conclude that careful screening for medications and underlying conditions predisposing to hypocalcemia is recommended to help prevent severe reactions due to citrate toxicity. Laboratory measurement of pre-procedure serum calcium levels in selected donors may identify cases requiring heightened vigilance. The case also illustrates the importance of maintaining preparedness for managing rare but serious reactions in volunteer apheresis blood donors. PMID- 17111420 TI - Comparison of plateletpheresis on the Fresenius AS.TEC 204 and Haemonetics MCS 3p. AB - This is an attempt at comparing two cell separators for plateletpheresis, namely the Fresenius AS.TEC 204 and Haemonetics MCS 3p, at a tertiary care center in India. Donors who weighed between 55-75 kg, who had a hematocrit of 41-43%, and platelet counts of 250x10(3)-400x10(3)/microl were selected for the study. The comparability of the donors who donated on the two cell separators were analysed by t-test independent samples and no significant differences were found (P>0.05). The features compared were time taken for the procedure, volume processed on the separators, adverse reactions of the donors, quality control of the product, separation efficiency of the separators, platelet loss in the donors after the procedure, and the predictor versus the actual yield of platelets given by the cell separator. The volume processed to get a target yield of >3x10(11) was equal to 2.8-3.2 l and equal in both the cell separators. Symptoms of citrate toxicity were seen in 4 and 2.5% of donors who donated on the MCS 3p and the AS.TEC 204, respectively, and 3 and 1% of donors, respectively, had vasovagal reactions. All the platelet products collected had a platelet count of >3x10(11); 90% of the platelet products collected on the AS.TEC 204 attained the predicted yield that was set on the cell separator where as 75% of the platelet products collected on the MCS 3p attained the target yield. Quality control of the platelets collected on both the cell separators complied with the standards except that 3% of the platelets collected on the MCS 3p had a visible red cell contamination. The separation efficiency of the MCS 3p was higher, 50-52% as compared to the 40-45% on the AS.TEC 204. A provision of double venous access, less adverse reactions, negligible RBC contamination with a better predictor yield of platelets makes the AS.TEC 204 a safer and more reliable alternative than the widely used Haemonetics MCS 3p. PMID- 17111421 TI - High-dose therapy and autologous stem-cell transplantation can improve event-free survival for indolent lymphoma: a study using patients as their own controls. AB - BACKGROUND: High-dose therapy (HDT) and autologous stem-cell transplantation (ASCT) remain controversial for indolent lymphoma patients. METHODS: The study was designed to evaluate the benefit of this strategy by retrospectively comparing for each patient the event-free survival (EFS) after ASCT with the duration of the disease phase just before the phase including ASCT (ie, the last qualifying phase, LQP). RESULTS: A total of 109 indolent lymphoma (mostly follicular lymphoma) patients were treated with HDT and ASCT. Before ASCT, patients experienced a median of 2 disease phases (range, 1-4). After a median 5 year follow-up from ASCT, overall survival was 67% and EFS was 43%. When each of the 92 patients experiencing recurrence was taken as her/his own control, EFS was longer after ASCT than the duration of LQP (62%, P < .01). During LQP, 86 patients (93%) experienced recurrence in less than 5 years, compared with only 58 (63%) who experienced recurrence in the 5 years after ASCT (P < .01). CONCLUSION: HDT and ASCT can significantly increase EFS in comparison with the duration of the LQP for indolent lymphoma patients and can change disease course. This methodology has been found useful for evaluating new strategies, especially with monoclonal antibodies. PMID- 17111422 TI - A model for targeted substitution mutagenesis during SOS replication of double stranded DNA containing cis-syn cyclobutane thymine dimers. AB - A model for ultraviolet mutagenesis is described that is based on the formation of rare tautomeric bases in pyrimidine dimers. It is shown that during SOS synthesis the modified DNA-polymerase inserts canonical bases opposite the dimers; the inserted bases are capable of forming hydrogen bonds with bases in the template DNA. SOS-replication of double-stranded DNA having thymine dimers, with one or both bases in a rare tautomeric conformation, results in targeted transitions, transversions, or one-nucleotide gaps. Structural analysis indicates that one type of dimer containing a single tautomeric base (TT*(1), with the "*" indicating a rare tautomeric base and the subscript referring to the particular conformation) can cause A:T --> G:C transition or homologous A:T --> T:A transversion, while another dimer (TT*(2)) can cause a one-nucleotide gap. The dimers containing T*(4) result in A:T --> C:G transversion, while TT*(5) dimers can cause A:T --> C:G transversion or homologous A:T --> T:A transversion. If both bases in the dimer are in a rare tautomeric form, then tandem mutations or double-nucleotide gaps can be formed. The dimers containing the rare tautomeric forms T*'(1) , T*'(2), T*'(3), T*'(4), and T*'(5) may not result in mutations. The question of whether dimers containing T*'(4) and T*'(5) result in mutations requires further investigation. PMID- 17111424 TI - Feasibility of conducting the micronucleus test in circulating erythrocytes from different mammalian species: an anatomical perspective. AB - The in vivo mammalian micronucleus test can be conducted easily on peripheral blood samples since the maturation of erythrocytes involves the loss of the major nucleus. In addition, mature erythrocytes are relatively long-lived, so that the test potentially can detect genotoxic damage caused by chronic exposures. However, some species have spleens that remove micronuclei from the peripheral circulation, making such measurements problematical. This report summarises haematological and mutagenesis studies dealing with this subject and provides an anatomical interpretation of the phenomenon. Anatomical features can be used to identify those species in which micronuclei are removed by the spleen. PMID- 17111423 TI - Polymorphisms in the thymidylate synthase promoter and the DNA repair genes XRCC1 and XPD in a Brazilian population. AB - Polymorphisms in genes responsible for maintaining genomic integrity are potential modifiers of disease risk. Since considerable interindividual and interethnic variation in DNA repair capacity has been associated with polymorphic alleles, we evaluated the frequency of the 2R/3R variants in the TS promoter, Arg194Trp and Arg399Gln in the XRCC1 gene, and Asp312Asn and Lys751Gln in the XPD gene in 364 healthy individuals from a Brazilian population separated by ethnicity (European ancestry and African ancestry). The genotypes were determined by PCR (TS) or by PCR-RFLP (XRCC1 and XPD). The frequency of the TS 3R allele was 0.56 for whites and 0.51 for nonwhites. In the case of the XRCC1 MspI polymorphism, the allele frequencies were 0.09 for 194Trp in both nonwhites and whites and 0.27 and 0.28 for 399Gln in nonwhites and whites, respectively. For the XPD 312Asn allele, we found a frequency of 0.25 in white individuals, which was significantly different (P = 0.025) from that seen in nonwhites (0.15). Similarly, the 751Gln polymorphic allele of the XPD gene was significantly more frequent (P < 0.002) in whites (0.30) than in nonwhites (0.20). The genotype frequencies were within Hardy-Weinberg equilibrium. We concluded that the genotype and allele frequencies of XPD gene polymorphism differed between white and nonwhite Brazilians, and that the frequencies of the XPD 312Asn and XRCC1 399Gln alleles in this Brazilian population showed ethnic variability when compared with those observed in other populations. PMID- 17111425 TI - Intraoral proliferative myositis: case report and literature review. AB - BACKGROUND: Proliferative myositis is a rare, benign, reactive intramuscular lesion of fibroblastic/myofibroblastic origin; an identical lesion in a subcutaneous or fascial location is referred to as proliferative fasciitis. The rapid growth rate and unusual histopathologic features have frequently been mistaken for a malignant process and have promoted unnecessary invasive procedures. Here we present only the third oral case of proliferative myositis, arising from the tongue of a 65-year-old man. METHODS AND RESULTS: Histologically, the resected lesion was composed of numerous fibroblastic or myofibroblastic spindle cells and variable numbers of large ganglion-like cells infiltrating between and around muscle fascicles, resembling a "checkerboard" configuration. A demographic profile of proliferative myositis of the head and neck is also provided, compiled from 19 patients culled from an English-language literature review and this report. CONCLUSIONS: Incisional biopsy or fine-needle aspiration biopsy of proliferative myositis of the head and neck should lead to spontaneous resolution and is, therefore, sufficient to render the diagnosis and to provide conservative treatment. Recurrence is extremely rare. PMID- 17111427 TI - Aberrant cytokine expression in serum of patients with adenoid cystic carcinoma and squamous cell carcinoma of the head and neck. AB - BACKGROUND: Squamous cell carcinoma (SCC) and adenoid cystic carcinoma (ACC) represent 2 clinically important subtypes of head and neck cancer. Our objective was to characterize and compare cytokine profiles in the systemic circulation of patients with SCC and ACC. METHODS: Multiplex analysis of 10 different cytokines (interleukin [IL]-1beta, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, granulocyte macrophage colony-stimulating factor [GM-CSF], interferon [IFN]-gamma, and tumor necrosis factor [TNF]-alpha) in the serum of patients with SCC (n = 20) and ACC (n = 20) and healthy controls (n = 20) was performed using the Luminex fluorescent-bead technology. RESULTS: Patients with SCC as well as patients with ACC showed an altered cytokine profile compared with healthy individuals. In patients with SCC, significantly elevated serum levels of the proinflammatory cytokines, IL-6 and IL-8, were observed. In patients with ACC, IL-8 serum levels were significantly elevated, and IL-6 serum levels were only increased in a subset of patients. CONCLUSIONS: A similar serum cytokine profile, with the predominance of proinflammatory cytokines, was observed in patients with SCC and ACC. The newly defined cytokine profile in ACC patients may form the basis for future investigations to explore the role of cytokines in ACC tumor progression and their potential value as predictive biomarkers. PMID- 17111428 TI - Use of single saphenous interposition vein graft for primary arterial circuit and secondary recipient site in head and neck reconstruction: a case report. AB - BACKGROUND: Autologous vein grafts are a valuable tool in microsurgical free tissue transfer. Interposition vein grafts offer the surgeon greater freedom when placing the free flap and choosing the recipient vessels, providing valuable options in case recipient vessels are not available for those patients with large wounds. Free flaps transferred to head and neck regions carry a higher risk of failure, which may be expected to increase more with the use of vein grafts. METHODS: We present our case with the double use of a single vein graft for both primary arterial conduit in end-to-end fashion and secondary end-to-side recipient site in the microsurgical reconstruction of a complicated head and neck defect. RESULTS: All these anastomoses and flaps survived perfectly, and the patient was discharged 14 days after the transfer of the second flap. CONCLUSION: Although the anastomosis of 2 flaps to a single vein graft was successful in our case, it represents a higher risk option than different recipient vessels. We provide this alternative procedure in selected patients, as there is no other receipt vessel or recipient blood flow strong enough to supply more than 1 flap. PMID- 17111429 TI - Evaluation of patterns of failure and subjective salivary function in patients treated with intensity modulated radiotherapy for head and neck squamous cell carcinoma. AB - BACKGROUND: Our aim was to correlate patterns of failure with target volume delineations in patients with head and neck squamous cell carcinoma (HNSCC) treated with intensity-modulated radiation therapy (IMRT) and to report subjective xerostomia outcomes after IMRT as compared with conventional radiation therapy (CRT). METHODS: Between January 2000 and April 2005, 69 patients with newly diagnosed nonmetastatic HNSCC underwent curative parotid-sparing IMRT at Stanford University. Sites included were oropharynx (n = 39), oral cavity (n = 8), larynx (n = 8), hypopharynx (n = 8), and unknown primary (n = 6). Forty-six patients received definitive IMRT (66 Gy, 2.2 Gy/fraction), and 23 patients received postoperative IMRT (60.2 Gy, 2.15 Gy/fraction). Fifty-one patients also received concomitant chemotherapy. Posttreatment salivary gland function was evaluated by a validated xerostomia questionnaire in 29 IMRT and 75 matched CRT patients >6 months after completing radiation treatment. RESULTS: At a median follow-up of 25 months for living patients (range, 10-60), 7 locoregional failures were observed, 5 in the gross target or high-risk postoperative volume, 1 in the clinical target volume, and 1 at the junction of the IMRT and supraclavicular fields. The 2-year Kaplan-Meier estimates for locoregional control and overall survival were 92% and 74% for definitive IMRT and 87% and 87% for postoperative IMRT patients, respectively. The mean total xerostomia questionnaire score was significantly better for IMRT than for CRT patients (p = .006). CONCLUSIONS: The predominant pattern of failure in IMRT-treated patients is in the gross tumor volume. Parotid sparing with IMRT resulted in less subjective xerostomia and may improve quality of life in irradiated HNSCC patients. PMID- 17111430 TI - Use of integrated 18F-FDG PET/CT to improve the accuracy of initial cervical nodal evaluation in patients with head and neck squamous cell carcinoma. AB - BACKGROUND: We investigated the accuracy of performing cervical nodal evaluation with using integrated (18)F-fluoro deoxyglucose positron emission tomography (PET)/CT for squamous cell carcinoma (SCC) of the head and neck as compared with using PET and contrast-enhanced CT (CECT) alone. METHODS: The presence of metastatic lymphadenopathy in each cervical nodal group (level I-VI) and the nodal (N) classification of 47 patients with SCC of the head and neck were determined by using PET, CECT, and PET/CT, respectively, and the results were verified according to the histopathologic findings. RESULTS: Among the 91 foci that had abnormal uptake on PET, the combined PET/CT images provided additional information over PET for the anatomical localization and lesion characterization of 18 sites (19.8%) in 17 patients (36.2%). PET/CT also showed the best results among the three imaging modalities for the sensitivity, specificity, and accuracy (91.8, 98.9, and 97.1%, respectively) for predicting metastatic nodes on a level by-level analysis, and PET/CT had a higher accuracy (85.1%) for the pathologic nodal classification over the clinical examinations (68.1%) or PET (70.2%). CONCLUSIONS: Combined PET/CT images are more accurate than the PET or CECT images alone for conducting cervical node evaluation in the patients suffering with head and neck SCC. PMID- 17111431 TI - Influence of social support on health-related quality of life outcomes in head and neck cancer. AB - BACKGROUND: Evidence that social support influences health-related quality of life (HRQOL) in oncologic patients could be particularly important for head and neck cancer because this disease can affect speech, eating, and facial aesthetics. METHODS: Multiple regression analyses were used in this prospective, observational study to determine the association between 394 patients' ratings of perceived post-treatment social support and HRQOL outcomes while controlling for possible confounding variables. RESULTS: Higher social support scores were significantly associated with higher scores in speech (p = .007), aesthetics (p = .015), social disruption (p = .045), and general mental health (p = .016) and with fewer depressive symptoms (p = .023) but not with general physical health (p = .191) or eating (p = .114). The magnitude of differences in the HRQOL outcomes for patients whose social support scores fell in the lowest and highest quartiles were clinically meaningful. CONCLUSIONS: Given the association between social support and HRQOL outcomes in this patient population, modification of perceived social support through clinical interventions could improve the survivorship of these patients. PMID- 17111432 TI - Acceleration of hyperfractionated chemoradiation regimen for advanced head and neck cancer. AB - BACKGROUND: Our aim was to evaluate the acceleration of a hyperfractionated, concurrent chemoradiation regimen (HxCRT) for advanced head and neck squamous cell carcinoma (HNSCC). METHODS: Patients with unresectable HNSCC were treated based on a previously published HxCRT regimen: 1.25 Gy twice daily to 70 Gy concurrent with cisplatin 12 mg/m(2)/day and 5-fluorouracil 600 mg/m(2)/day for 5 days, weeks 1, 5. This regimen was accelerated in this series by shortening the treatment from 7 to 6 weeks by omitting the planned mid-treatment 1-week break. RESULTS: Forty-six patients with T3-4/N3 disease were treated. The main acute toxicity was pharyngeal. Median weight change during therapy in patients with and without enteral feeding tubes was -3.8% and -7.9%, respectively (p = .08). Fifteen percent had late grade III pharyngeal toxicity. Local/regional and distant failure rates were 28% and 17%, respectively; 52% are alive without evidence of disease. CONCLUSIONS: In nonresectable HNSCC, acceleration of the HxCRT regimen is feasible, requiring enteral feeding tubes during therapy in most patients. PMID- 17111433 TI - Successful implant of long-term cryopreserved parathyroid glands after total parathyroidectomy. AB - BACKGROUND: Parathyroid cryopreservation is essential in some cases of parathyroid surgery. The fate of autografted tissue after long-term cryopreservation is not fully discussed in the literature. METHODS: The successful experience with the use of parathyroid tissues preserved for 21 months and 30 months is reported. RESULTS: Both patients were women with renal hyperparathyroidism who underwent total parathyroidectomy without autotransplantation. Patient 1 was a 40-year-old woman. At 21 months of follow up, her parathyroid hormone (PTH) level was undetectable, and despite oral calcium supplements, she was hypocalcemic. Forty-five cryopreserved fragments were thawed and implanted in her forearm. Calcium levels improved, and PTH steadily increased in both arms. PTH levels at 18 months after the autograft were 37.0 pg/mL in the contralateral arm and 1150.0 pg/mL in the implant arm. Patient 2 was a 44-year-old woman. After 30 months, her PTH was undetectable, and she underwent cryopreserved tissue implantation. CONCLUSION: These cases show that parathyroid tissue may remain viable even after long-term storage. PMID- 17111434 TI - Usefulness of MRI volumetric evaluation in patients with squamous cell cancer of the head and neck treated with neoadjuvant chemotherapy. AB - BACKGROUND: The purpose was to evaluate the efficacy of tumor volumetry on MRI as predictive of response to treatment with induction chemotherapy, comparing the results with endoscopy. METHODS: Fifty patients with advanced squamous cell carcinoma of the head and neck (SSCHN) who underwent MRI volumetry before and after neoadjuvant chemotherapy consisting of docetaxel, cisplatin, and 5 fluorouracil (TPF) were included in this study. The tumor volume was calculated by a slice-by-slice evaluation. With the standard software of a workstation, the area of the tumor was measured slice by slice using manual segmentation. To evaluate the efficacy of MRI volumetry, pretreatment volume was compared with pretreatment remission status as evaluated with endoscopy. RESULTS: Forty-five (90%) patients demonstrated a tumor downstaging after chemotherapy. Fourteen (28%) patients showed a complete histologic remission (CR), 31 (62%) patients showed a partial remission (PR). Pretreatment tumor volume was significantly different between patients whose tumor completely responded (CR) and those whose tumor did not completely respond or whose disease was stable or was progressive (p = .00023). We defined a threshold for the pretreatment tumor volume in patients with CR, which was equal to 29.71 cc. CONCLUSION: We propose that MRI tumor volume analyses can be a useful parameter to predict the response to neoadjuvant chemotherapy in SCCHN. PMID- 17111435 TI - Identification of proteins with high affinity for refolded and native PrPC. AB - PrPC, the cellular prion protein, is widely expressed in most tissues, including brain, muscle and the gastrointestinal tract, but its physiological role remains unclear. During propagation of transmissible spongiform encephalopathies (TSEs), prion protein is converted to the pathological isoform, PrPSc, in a process believed to be mediated by as-yet-unknown host factors. The identification of proteins associated with PrP may provide information about the biology of prions and the pathogenesis of TSEs. In the present work, we report proteins identified from brain tissue based on their ability to bind to recombinant PrP (recPrP) or form multimolecular complexes with native PrPC in the presence of cross-linkers. Immobilized his-tagged recPrP was used as an affinity matrix to isolate PrP interacting proteins from brain homogenates of normal individuals. In parallel, PrPC-associated proteins were characterized by cross-linking and co immunoprecipitation assays. The unknown molecules were identified by MS and the results of LC-MS/MS analysis were subsequently verified by Western blot. Both techniques resulted in identification of proteins participating in the formation of cytoskeleton and signal transduction, further supporting the hypothesis that PrP is involved in the organization and function of receptors throughout the nervous system. PMID- 17111437 TI - High-throughput analysis of mumps virus and the virus-specific monoclonal antibody on the arrays of a cationic polyelectrolyte with a spectral SPR biosensor. AB - We investigated the potential use of a spectral surface plasmon resonance (SPR) biosensor in a high-throughput analysis of mumps virus and a mumps virus-specific mAb on the arrays of a cationic polyelectrolyte, poly(diallyldimethylammonium chloride) (PDDA). The PDDA surface was constructed by electrostatic adsorption of the polyelectrolyte onto a monolayer of 11-mercaptoundecanoic acid (MUA). Poly-L lysine was also adsorbed onto the MUA monolayer and compared with the PDDA surface in the capacity of mumps virus immobilization. The PDDA surface showed a higher adsorption of mumps virus than the poly-L-lysine surface. The SPR signal caused by the virus binding onto the PDDA surface was proportional to the concentration of mumps virus from 0.5 x 10(5) to 14 x 10(5) pfu/mL. The surface structure of the virus arrays was visualized by atomic force microscopy. Then, a dose-dependent increase in the SPR signal was observed when various concentrations of the antimumps virus antibody in buffer or human serum were applied to the virus arrays, and their interaction was specific. Thus, it is likely that the spectral SPR biosensor based on the cationic polyelectrolyte surface may provide an efficient system for a high-throughput analysis of intact virus and serodiagnosis of infectious diseases. PMID- 17111436 TI - CyDye immunoblotting for proteomics: co-detection of specific immunoreactive and total protein profiles. AB - The development of ECL-Plex CyDye-conjugated secondary antibodies allows the advancement of conventional Western blotting, opening up possibilities for highly sensitive and quantitative protein confirmation and identification. We report a novel proteomic method to simultaneously visualise the total protein profile as well as the specific immunodetection of an individual protein species by combining cyanine CyDye pre-labelled proteins and antibody immunoblotting. This technique proposes to revolutionise both 2-D immunoprobing and protein confirmation following MS analysis. PMID- 17111438 TI - Proteome profiles of mucosal immunoglobulin uptake in inflamed porcine gut. AB - Acquisition of passive immunity by endocytosis of intact immunoglobulins (Ig) from colostrum is critical for prevention of intestinal and systemic diseases in neonatal mammals. We compared proteome patterns of healthy and inflamed gut tissues from pre-term piglets to investigate the effect of inflammation on acquisition of passive immunity. A clear difference in the two-dimensional gel electrophoresis protein patterns between healthy and inflamed intestinal tissues was observed, suggesting that inflamed tissues failed to absorb and transfer Ig from colostrum to epithelial cells. We have mapped and identified the Ig proteins that are taken up by healthy intestinal tissues, and found that isoforms of the IgA and IgG heavy chain and Ig kappa and lambda light chains were internalized. Our results indicate that colostrum protein uptake in the porcine gut is a selective process that is obstructed in inflamed pre-term gut. PMID- 17111439 TI - Beta-amyloid treatment of two complementary P301L tau-expressing Alzheimer's disease models reveals similar deregulated cellular processes. AB - Alzheimer's disease (AD) is characterized by Abeta peptide-containing plaques and tau-containing neurofibrillary tangles (NFTs). Both pathologies have been combined by crossing Abeta plaque-forming APP mutant mice with NFT-forming P301L tau mutant mice or by stereotaxically injecting beta-amyloid peptide 1-42 (Abeta42) into brains of P301L tau mutant mice. In cell culture, Abeta42 induces filamentous tau aggregates. To understand which processes are disrupted by Abeta42 in the presence of tau aggregates, we applied comparative proteomics to Abeta42-treated P301L tau-expressing neuroblastoma cells and the amygdala of P301L tau transgenic mice stereotaxically injected with Abeta42. Remarkably, a significant fraction of proteins altered in both systems belonged to the same functional categories, i.e. stress response and metabolism. We also identified model-specific effects of Abeta42 treatment such as differences in cell signaling proteins in the cellular model and of cytoskeletal and synapse associated proteins in the amygdala. By Western blotting (WB) and immunohistochemistry (IHC), we were able to show that 72% of the tested candidates were altered in human AD brain with a major emphasis on stress-related unfolded protein responsive candidates. These data highlight these processes as potentially important initiators in the Abeta42-mediated pathogenic cascade in AD and further support the role of unfolded proteins in the course of AD. PMID- 17111440 TI - Hydrothermal synthesis of molybdenum oxide based materials: strategy and structural chemistry. AB - The preparative flexibility of hydrothermal syntheses needs to be systemised for exploring complex structure-synthesis relationships and morphology control options in materials chemistry. This is demonstrated for the targeted hydrothermal preparation of molybdenum oxide materials: firstly, in situ studies were employed for the efficient production of MoO(3) nanofibres. Furthermore, ionic substances as structure-directing tools brought forward a new class of fluorinated polyoxomolybdates. PMID- 17111441 TI - Efficient enzymatic glycosylation of peptides and oligosaccharides from GalNAc and UTP. PMID- 17111442 TI - Liquid-crystalline and electron-deficient coronene oligocarboxylic esters and imides by twofold benzogenic Diels-Alder reactions on perylenes. AB - Alkyl esters, imides and imido-esters of coronene-tri-, -tetra- and octacarboxylic acids are accessible by a twofold oxidative benzogenic Diels-Alder reaction. Alkyl acrylates add to perylene, and maleic alkyl imides react twice with perylene as well as with perylene-tetracarboxylic tetraesters. Coronenes substituted with a greatly variable number of electron-withdrawing substituents are thus accessible, and di- and tetraimide derivatives are shown to be very pronounced electron-acceptor materials. The tri- and tetraalkyl esters and imidoesters self-assemble into columnar liquid-crystalline phases. PMID- 17111443 TI - De novo design and synthesis of helix-turn-helix structure from short and robust mixed helices derived from C-linked carbo-beta-amino acids. PMID- 17111444 TI - Sequence-specific DNA binding by noncovalent peptide-tripyrrole conjugates. PMID- 17111445 TI - A molecular solomon link. PMID- 17111447 TI - Heterogeneous copper-in-charcoal-catalyzed click chemistry. PMID- 17111446 TI - Lanthanitin: a chiral nanoball encapsulating 18 lanthanum ions by ferritin-like assembly. PMID- 17111448 TI - Lock-and-key principle on a microscopic scale: the case of the propylene oxide...ethanol complex. PMID- 17111449 TI - Relativistic functional groups: aryl carbon-gold bond formation by selective transmetalation of boronic acids. PMID- 17111451 TI - Rigid annulated carbon-sulfur structures. PMID- 17111450 TI - Is the ferric hydroperoxy species responsible for sulfur oxidation in cytochrome p450s? PMID- 17111452 TI - Mapping the landscape of potentially primordial informational oligomers: oligodipeptides and oligodipeptoids tagged with triazines as recognition elements. PMID- 17111453 TI - Mapping the landscape of potentially primordial informational oligomers: oligodipeptides tagged with 2,4-disubstituted 5-aminopyrimidines as recognition elements. PMID- 17111454 TI - Unfolding tin-cobalt interactions in oxide-based composite electrodes for Li-ion batteries by Mossbauer spectroscopy. AB - With a view to the development of new composite electrodes for lithium-ion batteries with electroactive tin and cobalt, Co-doped tin dioxide samples are studied. The role played by oxygen and cobalt atoms in the electrochemical behavior of tin-based electrodes for Li-ion batteries is examined by the powerful and selective (119)Sn Mossbauer spectroscopy. For the discharged electrodes, the oxygen atoms in the lithia matrix tend to destabilize the Sn(0) atoms. In contrast, the presence of cobalt atoms helps to form a matrix that stabilizes the reduced tin atoms. Cobalt-tin interactions in electrochemical reduced Co(x)Sn(1 x)O(2) electrodes are deduced from the electrochemical and Mossbauer results. PMID- 17111455 TI - Electrochemical microstructuring with short voltage pulses. AB - The application of short (nanosecond) voltage pulses between a tool electrode and a work piece immersed in an electrolyte solution allows the three-dimensional machining of electrochemically active materials with submicrometer resolution. The method is based on the finite charging time constant of the double-layer capacitance, which varies approximately linearly with the local separation between the electrode surfaces. Hence, the polarization of the electrodes during short pulses and subsequent electrochemical reactions are confined to regions where the electrodes are in sufficiently close proximity. This Minireview describes the principles behind electrochemical micro-structuring with short voltage pulses, and its current achievements and limitations. PMID- 17111456 TI - Release from silica SBA-3-like mesoporous fibers: cross-wall transport and external diffusion barrier. AB - The transport of guest molecules between adjacent pore channels (cross-wall transport) is the limiting factor in the release of guest molecules from SBA-3 like fibers. This specific mode of diffusion is identified by microscopic observation and studied quantitatively in a UV/Vis-monitored release experiment. Analysis of release curves reveals that the external particle surface offers resistance to the guest molecules passing through it (external diffusion barrier). This barrier is native to as-synthesized fibers and can be effectively modified to slow down the release. Extremely effective slowdown is achieved by deposition of a nanometer-thick layer of sodium silicate, that is, the guest molecules are then safely stored in the particles. PMID- 17111457 TI - Molecular dynamics of nanobubbles' collapse in ionic solutions. PMID- 17111458 TI - Dipole moment, hydrogen bonding and IR spectrum of confined water. PMID- 17111459 TI - Lack of effect of influenza immunisation on anticoagulant control in patients on long-term warfarin. AB - PURPOSE: The effect of influenza vaccination on oral anticoagulant control is uncertain but important to establish since anticoagulants are widely used and most patients taking them are candidates for immunisation because of age or underlying cardiac disease. We therefore prospectively evaluated the effect of influenza vaccination on International Normalised Ratio (INR) control in patients on long-term warfarin. METHODS: We undertook a prospective audit of patients on long-term warfarin attending a single hospital anticoagulant clinic who reported receiving influenza vaccination within the 10 days prior to a clinic visit. We compared the stability of anticoagulant control in the 12 months prior to and 10 days after immunisation, restricting analysis to those patients whose warfarin dosage was unchanged before and after vaccination. RESULTS: Of 106 consecutive patients who reported receiving influenza vaccination within the 10 days prior to a clinic visit, results were evaluable in 78 because the dose of warfarin was unchanged before and after vaccination. Influenza immunisation had no apparent effect on anticoagulant control. No bleeding or thrombotic complications were reported. CONCLUSIONS: Our findings suggest that it is not necessary to routinely monitor the INR more closely after influenza vaccination. PMID- 17111460 TI - A novel technique for visualization of the normal and cleft fetal secondary palate: angled insonation and three-dimensional ultrasound. AB - OBJECTIVE: To evaluate the effectiveness of a novel approach to the visualization of the fetal secondary palate using three-dimensional (3D) ultrasound. METHODS: Sonographic examinations were performed in normal fetuses and in one fetus with cleft lip and palate. To avoid acoustic shadowing from the alveolar ridge, the secondary palate was insonated at a 45 degrees angle in the sagittal plane, and 3D ultrasound was used to reconstruct axial and coronal planes. RESULTS: The secondary palate was successfully visualized in 10 of 15 normal fetuses, both in the axial and coronal planes. In the fetus with cleft lip and palate the lesion of the secondary palate was clearly demonstrated, particularly in the coronal plane. CONCLUSION: Angled insonation and 3D ultrasound allow clear visualization of normal and cleft secondary palate. PMID- 17111461 TI - Evaluation and comparison of 50 Hz current threshold of electrocutaneous sensations using different methods. AB - Leakage currents, tiny currents flowing from an everyday-life appliance through the body to the ground, can cause a non-adequate perception (called electrocutaneous sensation, ECS) or even pain and should be avoided. Safety standards for low-frequency range are based on experimental results of current thresholds of electrocutaneous sensations, which however show a wide range between about 50 microA (rms) and 1000 microA (rms). In order to be able to explain these differences, the perception threshold was measured repeatedly in experiments with test persons under identical experimental setup, but by means of different methods (measuring strategies), namely: direct adjustment, classical threshold as amperage of 50% perception probability, and confidence rating procedure of signal detection theory. The current is injected using a 1 cm2 electrode at the highly touch sensitive part of the index fingertip. These investigations show for the first time that the threshold of electrocutaneous sensations is influenced both by adaptation to the non-adequate stimulus and individual, emotional factors. Therefore, classical methods, on which the majority of the safety investigations are based, cannot be used to determine a leakage current threshold.The confidence rating procedure of the modern signal detection theory yields a value of 179.5 microA (rms) at 50 Hz power supply net frequency as the lower end of the 95% confidence range considering the variance in the investigated group. This value is expected to be free of adaptation influences, and is distinctly lower than the European limits and supports the stricter regulations of Canada and USA. PMID- 17111462 TI - Activity optimization method in SPECT: a comparison with ROC analysis. AB - A discriminant method for optimizing activity in nuclear medicine studies is validated by comparison with ROC (received operating characteristic)-curves. The method is tested in 21 single photon emission computerized tomography (SPECT), performed with a cardiac phantom. Three different lesions (L(1), L(2) and L(3)) were placed in the myocardium-wall by pairs for each SPECT. Three activities (84, 37 or 18.5 MBq) of 99mTc were used as background. Linear discriminant analysis was used to select the parameters that characterize image quality among the measured variables in the images [(Background-to-Lesion (B/L(i)) and Signal-to Noise (S(i)/N) ratios)]. Two clusters with different image quality (P=0.021) were obtained. The ratios B/L(1), B/L(2) and B/L(3) are the parameters used to construct the function with 100% of cases correctly classified into the clusters. The value of 37 MBq was the lowest tested activity for which good results for the B/L(i) ratios were obtained. The result coincides with the applied ROC-analysis (r=0.89). PMID- 17111463 TI - Acaricidal activities of some essential oils and their monoterpenoidal constituents against house dust mite, Dermatophagoides pteronyssinus (Acari: Pyroglyphidae). AB - The acaricidal activities of fourteen essential oils and fourteen of their major monoterpenoids were tested against house dust mites Dermatophagoides pteronyssinus. Five concentrations were used over two different time intervals 24 and 48 h under laboratory conditions. In general, it was noticed that the acaricidal effect based on LC(50) of either essential oils or monoterpenoids against the mite was time dependant. The LC(50) values were decreased by increasing of exposure time. Clove, matrecary, chenopodium, rosemary, eucalyptus and caraway oils were shown to have high activity. As for the monoterpenoids, cinnamaldehyde and chlorothymol were found to be the most effective followed by citronellol. This study suggests the use of the essential oils and their major constituents as ecofriendly biodegradable agents for the control of house dust mite, D. pteronyssinus. PMID- 17111464 TI - Targeted yield concept and a framework of fertilizer recommendation in irrigated rice domains of subtropical India. AB - Soil test crop response (STCR) correlation studies were carried out in Vindhyan alluvial plain during 2001 to 2004 taking IR-36 as test crop to quantify rice production in the context of the variability of soil properties and use of balanced fertilizers based on targeted yield concept. The soils were developed on gently sloping alluvial plain with different physiographic settings and notable variation in drainage condition. Soil properties show moderate variation in texture (loamy to clay), organic carbon content (4.4 to 9.8 g/kg), cation exchange capacity (10.2 to 22.4 cmol (p+)/kg) and pH (5.3 to 6.4). Soil fertility status for N is low to medium (224 to 348 kg/ha), P is medium to high (87 to 320 kg/ha) and K ranges from medium to high (158 to 678 kg/ha). Database regarding nutrient requirement in kg/t of grain produce (NR), the percent contribution from the soil available nutrients [CS (%)] and the percent contribution from the applied fertilizer nutrients [CF (%)] were computed for calibrating and formulating fertilizer recommendations. Validity of the yield target for 7 and 8 t/ha was tested in farmers' fields and yields targets varied at less than 10%. The percent achievement of targets aimed at different level was more than 90%, indicating soil test based fertilizer recommendation approach was economically viable within the agro-ecological zone with relatively uniform cropping practices and socio-economic conditions. PMID- 17111465 TI - Regularities in the E. coli promoters composition in connection with the DNA strands interaction and promoter activity. AB - The energy of interaction between DNA strands in promoters is of great functional importance. Visualization of the energy of DNA strands distribution in promoter sequences was achieved. The separation of promoters in groups by their energetic properties enables evaluation of the dependence of promoter strength on the energetic properties. The analysis of groups (clusters) of promoters distributed by the energy of DNA strands interaction in -55, -35, -10 and +6 sequences indicates their connection with the transcriptional activity. PMID- 17111466 TI - Quality of effluents from Hattar Industrial Estate. AB - Of 6634 registered industries in Pakistan, 1228 are considered to be highly polluting. The major industries include textile, pharmaceutical, chemicals (organic and inorganic), food industries, ceramics, steel, oil mills and leather tanning which spread all over four provinces, with the larger number located in Sindh and Punjab, with smaller number in North Western Frontier Province (NWFP) and Baluchistan. Hattar Industrial Estate extending over 700 acres located in Haripur district of NWFP is a new industrial estate, which has been developed with proper planning for management of industrial effluents. The major industries located in Hattar are ghee industry, chemical (sulfuric acid, synthetic fiber) industry, textile industry and pharmaceuticals industry. These industries, although developed with proper planning are discharging their effluents in the nearby natural drains and ultimately collected in a big drain near Wah. The farmers in the vicinity are using these effluents for growing vegetables and cereal crops due to shortage of water. In view of this discussion, there is a dire need to determine if these effluents are hazardous for soil and plant growth. So, effluents from different industries, sewage and normal tap water samples were collected and analysed for pH, electrical conductivity (EC), total soluble salts (TSS), biological oxygen demand (BOD), chemical oxygen demand (COD), total nitrogen, cations and anions and heavy metals. The effluents of ghee and textile industries are highly alkaline. EC and TSS loads of ghee and textile industries are also above the National Environmental Quality Standards (NEQS), Pakistan. All the effluents had residual sodium carbonates (RSCs), carbonates and bicarbonates in amounts that cannot be used for irrigation. Total toxic metals load in all the effluents is also above the limit i.e. 2.0 mg/L. Copper in effluents of textile and sewage, manganese in ghee industry effluents and iron contents in all the effluents were higher than NEQS. BOD and COD values of all the industries are also above the NEQS. On the whole, these effluents cannot be used for irrigation without proper treatment otherwise that may cause toxicity to soil, plants and animals as well add to the problems of salinity and sododicity. Similarly, these effluents cannot be used for fish farming. PMID- 17111467 TI - Imbalanced free radicals and antioxidant defense systems in schizophrenia: a comparative study. AB - OBJECTIVE: To examine changes of blood oxidative-antiovidative level in schizophrenic patients and its relationship with clinical symptoms. METHODS: Forty-six Chinese patients met DSM-IV (Diagnostic and Statistical Manual of Mental Disorders-IV) criteria for schizophrenia and fifty age- and sex-matched healthy controls were enrolled in the present study. Baseline psychiatric symptom severity was assessed with brief psychiatric rating scale, positive and negative syndrome scale on the blood draw day. Fresh blood samples were collected to measure levels of nitric oxide and lipid peroxide in plasma as well as activities of superoxide dismutase, catalase and glutathione peroxidase in red blood cells by spectrophotometric assays simultaneously. RESULTS: Comparison of the biochemical parameters indicated that the level of nitric oxide and lipid peroxide increased in patient group, which represented a positive correlation with positive scale scores; while the activities of three critical enzymes decreased and showed a negative linear correlation. CONCLUSION: This study showed that there are dysregulation of free radical metabolism and poor activities of the antioxidant defense systems in schizophrenic patients. Excess free radicals formation may play a critical role in the etiology of schizophrenia. Using antioxidants might be an effective therapeutic approach to partially alleviate or prevent the symptoms of schizophrenia. PMID- 17111468 TI - A probiotic treatment containing Lactobacillus, Bifidobacterium and Enterococcus improves IBS symptoms in an open label trial. AB - OBJECTIVE: To evaluate the efficacy and safety of live combined Bifidobacterium, Lactobacillus and Enterococcus capsules in treatment of irritable bowel syndrome. METHODS: Eighty-five patients [male 32, female 53; age (45.31+/-11.72) years] were given live combined Bifidobacterium, Lactobacillus and Enterococcus capsules 1260 mg/d t.i.d.x4 weeks. Syndrome scales were used to evaluate the efficacy in gastrointestinal syndrome. Fecal flora was also measured before and after the treatment. Six bacteria were cultured and the colony forming units were counted in stool. SPSS was used for data analysis. RESULTS: Seventy-four patients finished the follow-up. No side-effect was found. For treatment of irritable bowel syndrome, the effective rate of live combined Bifidobacterium, Lactobacillus and Enterococcus capsules was 56.8% in the second week, 74.3% in the fourth week and 73.0% in the sixth week. Single symptom was improved, especially in abdominal pain and stool character. The probiotica containing live combined Bifidobacterium, Lactobacillus and Enterococcus could increase bifidobacterium count (P<0.01) and lactobacillus count (P<0.05); decrease bacteroides count (P<0.05) and enterococci count (P<0.01); No obvious changes were observed in clostridium difficile colonitis and enterobacteriaceae (P>0.05). CONCLUSION: The result of the study indicated that the administration of live combined Bifidobacterium, Lactobacillus and Enterococcus improved the symptom of irritable bowel syndrome and that there was a gradual increase of this effect. Thereafter conditions remained stable for 2 weeks. That improvement may be associated with alterations in gastrointestinal flora. PMID- 17111469 TI - Dose-response study of spinal hyperbaric ropivacaine for cesarean section. AB - BACKGROUND: Spinal hyperbaric ropivacaine may produce more predictable and reliable anesthesia than plain ropivacaine for cesarean section. The dose response relation for spinal hyperbaric ropivacaine is undetermined. This double blind, randomized, dose-response study determined the ED50 (50% effective dose) and ED95 (95% effective dose) of spinal hyperbaric ropivacaine for cesarean section anesthesia. METHODS: Sixty parturients undergoing elective cesarean section delivery with use of combined spinal-epidural anesthesia were enrolled in this study. An epidural catheter was placed at the L1 approximately L2 vertebral interspace, then lumbar puncture was performed at the L3 approximately L4 vertebral interspace, and parturients were randomized to receive spinal hyperbaric ropivacaine in doses of 10.5 mg, 12 mg, 13.5 mg, or 15 mg in equal volumes of 3 ml. Sensory levels (pinprick) were assessed every 2.5 min until a T7 level was achieved and motor changes were assessed by modified Bromage Score. A dose was considered effective if an upper sensory level to pin prick of T7 or above was achieved and no intraoperative epidural supplement was required. ED50 and ED95 were determined with use of a logistic regression model. RESULTS: ED50 (95% confidence interval) of spinal hyperbaric ropivacaine was determined to be 10.37 (5.23 approximately 11.59) mg and ED95 (95% confidence interval) to be 15.39 (13.81approximately 23.59) mg. The maximum sensory block levels and the duration of motor block and the rate of hypotension, but not onset of anesthesia, were significantly related to the ropivacaine dose. CONCLUSION: The ED50 and ED95 of spinal hyperbaric ropivacaine for cesarean delivery under the conditions of this study were 10.37 mg and 15.39 mg, respectively. Ropivacaine is suitable for spinal anesthesia in cesarean delivery. PMID- 17111470 TI - Effect of glycine site/NMDA receptor antagonist MRZ2/576 on the conditioned place preference and locomotor activity induced by morphine in mice. AB - OBJECTIVE: To study the effect of glycine site/NMDA (N-methyl-D-aspartate) receptor antagonist MRZ2/576 on the conditioned place preference (CPP) and locomotor activity induced by morphine in mice. METHODS: Different doses (1.25, 2.5 and 5 mg/kg, i.p.) of MRZ2/576 were used to evaluate the effect of MRZ2/576 on the acquisition and expression of CPP induced by morphine (5 mg/kg) in mice. In addition, we examined the locomotor activity of mice in conditioning and testing phase of CPP paradigm. RESULTS: MRZ2/576 alone could not establish place preference, but a 5 mg/kg dose of MRZ2/576 could block both acquisition and expression of morphine-induced CPP. In testing phase of CPP, there was no statistical difference for locomotor activity between the groups; injection of MRZ2/576 showed a dose-dependent decrease of locomotor activity on both control and morphine-treated mice, especially 5 mg/kg of MRZ2/576 significantly suppressed the locomotor activity of mice. CONCLUSION: Based on the present results, we assume that MRZ2/576 can antagonize the rewarding effect of morphine, suggesting that this glycine site/NMDA receptor antagonist could be used to treat addictions due to its light side effect profile. PMID- 17111471 TI - Anticancer effects of Chinese herbal medicine, science or myth? AB - Currently there is considerable interest among oncologists to find anticancer drugs in Chinese herbal medicine (CHM). In the past, clinical data showed that some herbs possessed anticancer properties, but western scientists have doubted the scientific validity of CHM due to the lack of scientific evidence from their perspective. Recently there have been encouraging results, from a western perspective, in the cancer research field regarding the anticancer effects of CHM. Experiments showed that CHM played its anticancer role by inducing apoptosis and differentiation, enhancing the immune system, inhibiting angiogenesis, reversing multidrug resistance (MDR), etc. Clinical trials demonstrated that CHM could improve survival, increase tumor response, improve quality of life, or reduce chemotherapy toxicity, although much remained to be determined regarding the objective effects of CHM in human in the context of clinical trials. Interestingly, both laboratory experiments and clinical trials have demonstrated that when combined with chemotherapy, CHM could raise the efficacy level and lower toxic reactions. These facts raised the feasibility of the combination of herbal medicines and chemotherapy, although much remained to be investigated in this area. PMID- 17111472 TI - Harvard Men's Health Watch at 10 years: a special anniversary issue. PMID- 17111473 TI - Cholesterol. PMID- 17111474 TI - Benign prostatic hyperplasia (BPH). PMID- 17111475 TI - Hypertension. PMID- 17111476 TI - New scans. PMID- 17111477 TI - Atherosclerosis, inflammation, and C-reactive protein. PMID- 17111478 TI - Treating prostate cancer. PMID- 17111479 TI - Tylenol safety: is there reason to worry? PMID- 17111480 TI - Poison safety. Warnings about accidental poisonings: Triaminic Vapor Patch...and WellPatch Cough and Cold Soothing Vapor Pads...warning about alternative treatment for Lyme disease. PMID- 17111481 TI - Infections/illnesses. Whooping cough--more common than we think...and measles is still with us. PMID- 17111482 TI - Benadryl for infant sleep? PMID- 17111483 TI - Child safety. Shopping carts: a source of injuries to children. PMID- 17111484 TI - Product recalls. Recalls: children's lamp sold at The Home Depot...and girls' hooded sweatshirts with drawstring recalled for strangulation hazard. PMID- 17111485 TI - Routine HIV screening could be key to finding the unknown infected. Counseling, separate informed consent no longer needed. PMID- 17111486 TI - Michigan devises plan for routine testing in ERs. Program find 50 percent more new HIV cases. PMID- 17111487 TI - Investigators find genetics help explain HIV dementia. Finding is a clue, but not entire answer. PMID- 17111488 TI - Hospital program reduces ART errors by 50 percent. Adherence improves as a result. PMID- 17111489 TI - ADAP problems continue as waiting lists top 300. More people who need drugs are off the radar. PMID- 17111490 TI - Palliative care model lost in today's HIV/AIDS care. Comfort care and pain control still necessary. PMID- 17111491 TI - Adherence strategies. FDA approves ddI generic version. PMID- 17111492 TI - HIV prophylaxis still has no clear cut answer. PMID- 17111493 TI - CDC's global AIDS outlook gives preview of WHO report. Prevalence declines in some hot spots. PMID- 17111494 TI - A perturbation method for predicting the temperature and stress sensitivities of quartz vibrating structures simulated by finite-element analysis. AB - Thermal and mechanical sensitivities of vibrating structures and wave guides are key parameters for the optimization of high stability resonant devices operating in the ultrasonic frequency range (from a few tenth of kilohertz to a few gigahertz). In this paper, the possibility to simulate and predict temperature coefficients of frequency (TCF) of quartz transducers of any shape as well as their stress sensitivity coefficients is addressed. The theoretical developments based on harmonic finite-element analysis coupled with a variational perturbation method are detailed, showing how to derive the regarded parameters. The proposed approach is validated using a two-dimensional (2-D) model of a plane face-bulk acoustic resonator for which an analytical model can give access to both TCF and stress sensitivity coefficients. It is then applied to a 2-D model of convex plane bulk acoustic resonator of singly rotated quartz and used to compute the first order TCF of a 3-D model of a tuning fork structure. In the latter case, the importance of considering the actual excitation of the device is demonstrated, allowing for the accurate definition of angular loci for which thermal compensation can be expected, in agreement with literature. Possible extensions and improvements of the proposed method is discussed in conclusion. PMID- 17111495 TI - [Quality of life: attitudes and perspectives of doctors in a thoracic oncology regional care network]. AB - BACKGROUND: The evaluation of health-related quality of life (HRQOL) of lung cancer patients is significant given their poor prognosis. Consequently, HRQOL is often currently assessed throughout clinical trials and sometimes in daily medical practice. The objective here was to determine physicians' perspectives on evaluating HRQOL. MATERIAL AND METHODS: The team carried out a prospective study of physicians belonging to a thoracic oncology network using an original questionnaire of 33 items. RESULTS: 102 physicians were questioned and 51 responded. Physicians' knowledge on HRQOL was frequently of a low level and incomplete, but most of them considered HRQOL as important HRQOL assessment in daily practice was often done intuitively. Many physicians noted that they lacked adequate time to explore and integrate all of the fields which should be covered by HRQOL in their clinical practice, but they felt overall that the HRQOL study improved doctor/patient communication and interaction. Physicians described an "ideal" HRQOL questionnaire as one containing 10-15 items within 6 domains (physical, psychological, general health, respiratory symptoms, family and social) and which could be completed in 5-8 minutes. In addition, such a questionnaire might integrate an opportunity for patients to isolate and identify items and issues that they consider to be most important to them. CONCLUSIONS: Physicians' will to consider and integrate the evaluation of HRQOL in the management of lung cancer patients is strong but meets some structural challenges (related to HRQOL assessment and interpretation of data) but also conjectural difficulties (with respect to a lack of knowledge). Training efforts are needed as well as the development of easier tools to facilitate the evaluation of HRQOL. PMID- 17111496 TI - The road not taken: how tribes choose between tribal and Indian Health Service management of health care services. PMID- 17111497 TI - Appropriate technologies in the traditional Native American smokehouse: public health considerations in tribal community development. PMID- 17111498 TI - Drought as a factor in the Jamestown colony, 1607-1612. PMID- 17111499 TI - Genetic discrimination in health insurance--comprehensive legal solutions for a (not so) special problem? PMID- 17111500 TI - Hospital chaplaincy under the HIPAA Privacy Rule: health care or "just visiting the sick"? PMID- 17111501 TI - Creating human embryos for research: a scientist's perspective on managing the legal and ethical issues. PMID- 17111502 TI - Of pills and needs: involuntarily medicating the psychotic inmate when execution looms. PMID- 17111503 TI - Prenatal testing gone awry: the birth of a conflict of ethics and liability. PMID- 17111504 TI - A chimera in every sense: standard of care for physicians practicing complementary and alternative medicine. PMID- 17111505 TI - Patient safety or profit: what incentives are blood shield laws and FDA regulations creating for the tissue banking industry? PMID- 17111506 TI - Slave demography in the Lowcountry, 1670-1740: from frontier society to plantation regime. PMID- 17111507 TI - The accidental terrorists. PMID- 17111508 TI - Tools you can trust. PMID- 17111509 TI - Must physicians respect an incompetent patient's refusal of treatment? PMID- 17111510 TI - Choosing paternalism? PMID- 17111511 TI - In the line of duty: SARS and physician responsibility in epidemics. PMID- 17111512 TI - How much suffering is enough? PMID- 17111513 TI - Do drug firm links sway psychiatry? PMID- 17111514 TI - Scandal grows over suspect body parts. PMID- 17111515 TI - One of a kind: what did Dolly the sheep really do for us? PMID- 17111516 TI - Ten years on, has the cloning dream died? PMID- 17111517 TI - Therapeutic cloning set back by hype and fraud. PMID- 17111518 TI - 'Virgin birth' stem cells bypass ethical objections. PMID- 17111519 TI - Clarification: Information Accuracy and Truthfulness Policy, Requirement 7. PMID- 17111520 TI - No need for monkeys. PMID- 17111521 TI - Bioethics centers and committees: cultural origins and current status. PMID- 17111522 TI - Dispensing medical marijuana: some halachic parameters. PMID- 17111523 TI - The negotiation and development of a clinical trial agreement. AB - A carefully negotiated clinical trial agreement will protect a biotechnology and medical device company in critical areas, including the management and control of data and confidential information, defining publication rights and the ownership of intellectual property. Properly constructed, the agreement can increase a company's value. This article will review the major provisions of a standard industry sponsored clinical trial agreement; the principle areas where there is likely to be differing views that need to be reconciled and interrelated ethical concerns. A form of clinical trial agreement designed for a single site therapeutic trial in the United States follows the article. PMID- 17111524 TI - The legal status of embryos and implications for reproductive technologies and biotechnology research. AB - The legal status of embryos in American law is changing. At present, most states do not afford embryos the same protections as a born person, but some states are attempting to change this standard. Granting embryos the same legal status as born human beings poses a significant problem for industries that work with embryos, especially fertility treatment facilities and scientists researching stem cell and gene therapy technologies. This paper describes the methods of defining embryos in American law, and discusses the implications of granting embryos the same rights as born persons for the reproductive technology and scientific research industries. PMID- 17111525 TI - Financial conflicts of interest in biomedical human subject research. AB - The purpose of this paper is to examine the past, present and future of financial conflict of interest regulation in biomedical human subject testing. Part I will briefly review the forces giving rise to the current controversy. Part II will examine the more influential ethical codes on human subject testing and argue that they are inconclusive on the subject of financial conflicts of interest. Part III will examine the various regulations now in place and identify their serious flaws. Part IV will critique the leading proposals for reform. The Conclusion will synthesize the best features of the various proposals for reform and suggest improvements left unaddressed by these proposals. PMID- 17111526 TI - The bounds of science and ethics. PMID- 17111527 TI - National Institutes of Health enacts final ethics regulations. AB - NIH new ethics regulations limit its employees activities, investments, and receipt of awards of NIH employees. PMID- 17111528 TI - Research examines the minimal risk standard for pediatric research. AB - The "minimal" risk standard for protection of pediatric research subjects often fails due to unclear definitions and uneven IRB application, requiring an alternative standard to better assist pediatric research assessment. PMID- 17111529 TI - Face transplants and the difficulties of obtaining research approval. AB - After over one year of consideration, the world's first face transplant has obtained research approval despite ethical and safety concerns. PMID- 17111530 TI - Stem cell research debate shifting to the courts. AB - California stem cell initiative faces legal challenges from groups opposing embryonic research and abortion opponents. PMID- 17111531 TI - Agency proposes safeguards for children in clinical trials. PMID- 17111532 TI - When informed consent is not required in studies. PMID- 17111533 TI - Case study: university's defenses against federal noncompliance charges begin to crumble. PMID- 17111534 TI - Hospice utilization: political, cultural, and legal issues. AB - The provision of end-of-life care is considered a substantive part of nursing, and hospice is often considered the "gold standard" of optimal end-of-life care. Unfortunately, however, only an estimated 43% of patients eligible for hospice actually receive hospice services (Harrison, Ford, & Wilson, 2005). The purpose of this article is to describe the political cultural, and legal issues associated with the underutilization of hospice care. Specifically, this article will outline the policy issues for accessing hospice under Medicare and Medicaid, the cultural and social issues in hospice utilization, and the legal implications of these issues. In addition, this article will serve to offer initial solutions to the problems at hand in order to guide nurses in promoting effective, efficient, and realized access to hospice services. PMID- 17111535 TI - Major changes in benefit design: a plausible way to control costs? PMID- 17111536 TI - Enhanced bronchial expression of extracellular matrix proteins in chronic obstructive pulmonary disease. AB - Remodeling of airways and blood vessels is an important feature in chronic obstructive pulmonary disease (COPD). By using immunohistochemical analysis, we examined bronchial expression patterns of various extracellular matrix (ECM) components such as collagens (subtypes I, III, and IV), fibronectin, and laminin beta2 in patients with COPD (forced expiratory volume in 1 second [FEV1] or=85%; n = 16) and correlated expression data with lung function. Quantitative analysis revealed enhanced levels (P < .01) of total collagens I, III, and IV in surface epithelial basement membrane (SEBM) and collagens I and III in bronchial lamina propria (P < .02) and adventitia (P < .05) in COPD. Distinct and increased (P < .05) vascular expression of fibronectin accounts for intimal vascular fibrosis, whereas laminin beta2 (P < .05) was elevated in airway smooth muscle (ASM). FEV1 values inversely correlated with collagens in the SEBM, fibronectin in bronchial vessels, and laminin in the ASM. Our data suggest that COPD exhibits increased bronchial deposition of ECM proteins that contribute to deteriorated lung function and airway remodeling. PMID- 17111537 TI - The identification of ganglion cells in Hirschsprung disease by the immunohistochemical detection of ret oncoprotein. PMID- 17111538 TI - Only time can tell: unethical research and the passage of time. PMID- 17111539 TI - Singleton v. Norris: precursor to Abu Ghraib? The importance of role integrity in medicine. PMID- 17111540 TI - Defining the limits of conscientious objection in health care. PMID- 17111541 TI - Frozen embryo adoption and the United States government. PMID- 17111542 TI - Updating Medicare's physician fees: the sustainable growth rate methodology. AB - Medicare's method to annually update the fees it pays physicians has been under fire for some time--specifically, since the method determined that physician fees should be reduced rather than increased. The update method, called the sustainable growth rate (SGR), was implemented to control the growth in Medicare physician spending. Yet Congress, in response to physician concerns about beneficiary access to care, has acted to avert physician fee cuts since 2003. Although this signals dissatisfaction with the SGR methodology, there is yet to be a widely accepted physician fee update proposal that balances federal budgetary realities with the need to ensure beneficiary access. And the cost of changing the update method continues to mount, adding to the difficulties of developing a solution that meets the needs of all stakeholders. This issue brief describes the SGR methodology, the reasons why projected physician fee updates are negative, and some options that have been proposed to remedy the current situation. This issue brief is the second of two related papers on physician spending and Medicare's sustainable growth rate methodology. The companion paper was published on October 9, 2006 (see Issue Brief 815, available at www.nhpf.org/pdfs_ib/IB815_PhysicianSpending_10-09-06.pdf). PMID- 17111543 TI - Judaism and abortion: the hijacking of a tradition. PMID- 17111544 TI - China's one-child policy: twenty-five years later. PMID- 17111545 TI - Dementia and the ethics of decision-making. PMID- 17111546 TI - "The history of us": social science, history, and the relations of family in Canada. PMID- 17111547 TI - Visually impaired (VI) education in Romania and the United Kingdom: special education in Romania since 1990 for blind and partially sighted children and young people, with comparisons drawn from similar experiences of legislative and educational changes in England and Wales since 1981. AB - This article reviews the educational facilities available in the last decades of the 20th century for blind and partially sighted children and young people in Romania and the UK. A turning point in England and Wales came with the Education (Special Educational Needs) Act 1981. For Romania, the turning point came in 1990 with the overthrow of the Ceaucescu regime in late 1989. Romania is seeking entry to the European Union in 2007. This has provided an incentive to bring special educational philosophy and facilities quickly into line with other countries in the European Union. Similar problems regarding inclusion are encountered in both countries. Differentiating features are their different demographic characteristics and the educational resources and assets available at the time of the major changes. Finally, there is a brief review of the number of people with sight loss in paid employment as the next logical focal point for a national inclusion strategy. The UK has been able to develop and advance further than Romania in this sphere, but the legal framework is in place in Romania. The generic terms visually impaired (VI) and sight loss are both used throughout when referring to blind and partially sighted people. PMID- 17111548 TI - Impact of spine surgery on signs and symptoms of spinal deformity. AB - Paediatric scoliosis is associated with signs and symptoms including reduced pulmonary function, increased pain and impaired quality of life, all of which worsen during adulthood, even when the curvature remains stable. Spinal fusion has been used as a treatment for nearly 100 years. In 1941, the American Orthopedic Association reported that for 70% of patients treated surgically, outcome was fair or poor: an average 65% curvature correction was reduced to 27% at >2 year follow-up and the torso deformity was unchanged or worse. Outcome was worse in children treated surgically before age 10, despite earlier intervention. Today, a reduced magnitude of curvature obtained by spinal fusion in adolescence can be maintained for decades. However, successful surgery still does not eliminate spinal curvature and it introduces irreversible complications whose long-term impact is poorly understood. For most patients there is little or no improvement in pulmonary function. Some report improved pain after surgery, some report no improvement and some report increased pain. The rib deformity is eliminated only by rib resection which can dramatically reduce respiratory function even in healthy adolescents. Outcome for pulmonary function and deformity is worse in patients treated surgically before the age of 10 years, despite earlier intervention. Research to develop effective non-surgical methods to prevent progression of mild, reversible spinal curvatures into complex, irreversible structural deformities, is long overdue. PMID- 17111549 TI - Rehabilitation services for children: therapists' perceptions. AB - The objective of this study was to examine occupational therapists' (OT) and physiotherapists' (PT) perceptions regarding waiting time and the quality and quantity of the services they provide for children with disabilities. A survey was sent by post to all paediatric OTs and PTs in Quebec, Canada. A Service Delivery Questionnaire included questions regarding therapist/client characteristics, waiting times and quality and quantity of services provided. The Measure of Processes of Care for Service Providers (MPOC-SP) rated use of family centred care by responding clinicians. Waiting times were longest for OT services (p<0.0001), speech/language interventions (p<0.0001) and services in rehabilitation and community health centres (p<0.0001). Quality of services was rated higher by PTs, experienced therapists and those using more methods of keeping up-to-date. The frequency and duration of services varied according to profession, type of clientele and setting. According to clinicians directly involved in the provision of rehabilitation services, long delays exist for children waiting for rehabilitation services, and perceived quality of services differs according to specific therapist and client characteristics. These findings will assist those involved in planning the distribution of co-ordinated rehabilitation services for children. PMID- 17111550 TI - [Sciences of power, power of sciences, several trends in the Canadian historiography of sciences]. PMID- 17111551 TI - UK physicians' attitudes and practices in long-term non-invasive ventilation of Duchenne Muscular Dystrophy. AB - Previous studies have shown that long-term non-invasive ventilation (NIV) is not always routinely offered by all physicians in Duchenne Muscular Dystrophy (DMD), despite evidence that this treatment improves quality of life and survival. This study examined UK physicians' practices related to respiratory follow-up and DMD ventilation. A mailed questionnaire was used. Thirty-eight of the 59 (64%) UK physicians identified via the Muscular Dystrophy Campaign (MDC) responded. Eighty one per cent of respondents felt ethically obliged to discuss NIV with families while 13% believed that NIV results in poor quality of life. Forty-seven per cent of physicians discuss in-depth the use of NIV when the patient is in respiratory failure. Eighty-four ventilated DMD patients in the respondents' practice use NIV (via Bi-Pap Nasal mask). Nearly 66% of physicians do not consider the public cost to be an impediment to offering NIV, despite significant problems with resources' allocation in their area. While the majority of UK physicians have comparable attitudes and practices regarding NIV, the questionnaire highlighted that not all specialists were aware of the existence of consensus guidelines regarding respiratory monitoring. In addition, different practices of disclosure of life prolonging ventilation options were used by different physicians. Seventy-one per cent of physicians wished for national consensus guidelines for different DMD age groups. PMID- 17111552 TI - Single parenthood in the past: Canadian census categories, 1891-1951, and the "normal" family. PMID- 17111553 TI - Contested bodies: property models and the commodification of human biological artefacts. PMID- 17111554 TI - Feminists on the inalienability of human embryos. AB - The feminist literature against the commodification of embryos in human embryo research includes an argument to the effect that embryos are "intimately connected" to persons, or morally inalienable from them. We explore why embryos might be inalienable to persons and why feminists might find this view appealing. But, ultimately, as feminists, we reject this view because it is inconsistent with full respect for women's reproductive autonomy and with a feminist conception of persons as relational, embodied beings. Overall, feminists should avoid claims about embryos' being inalienable to persons in arguments for or against the commodification of human embryos. PMID- 17111555 TI - Reproductive freedom, self-regulation, and the government of impairment in utero. AB - This article critically examines the constitution of impairment in prenatal testing and screening practices and various discourses that surround these technologies. While technologies to test and screen (for impairment) prenatally are claimed to enhance women's capacity to be self-determining, make informed reproductive choices, and, in effect, wrest control of their bodies from a patriarchal medical establishment, I contend that this emerging relation between pregnant women and reproductive technologies is a new strategy of a form of power that began to emerge in the late eighteenth century. Indeed, my argument is that the constitution of prenatal impairment, by and through these practices and procedures, is a widening form of modern government that increasingly limits the field of possible conduct in response to pregnancy. Hence, the government of impairment in utero is inextricably intertwined with the government of the maternal body. PMID- 17111556 TI - Adoption, ART, and a re-conception of the maternal body: toward embodied maternity. AB - We criticize a view of maternity that equates the natural with the genetic and biological and show how such a practice overdetermines the maternal body and the maternal experience for women who are mothers through adoption and ART (Assisted Reproductive Technologies). As an alternative, we propose a new framework designed to rethink maternal bodies through the lens of feminist embodiment. Feminist embodied maternity, as we call it, stresses the particularity of experience through subjective embodiment. A feminist embodied maternity emphasizes the physical relations of the subjective lived-body rather than the genetic or biological connections. Instead of universalizing claims about the maternal body, embodied maternity looks to communicable experiences and empathetic understanding. PMID- 17111557 TI - ECOG: CHAARTED--ChemoHormonal therapy versus androgen ablation randomized trial for extensive disease in prostate cancer. PMID- 17111558 TI - NSABP B-39, RTOG 0413: A Randomized Phase III Study of conventional whole breast irradiation versus partial breast irradiation for women with stage 0, I, or II breast cancer. PMID- 17111559 TI - Bevacizumab in combination with 5-fluorouracil-based chemotherapy in the second line treatment of metastatic colorectal cancer. PMID- 17111560 TI - Exit strategies in Europe. PMID- 17111561 TI - Randomization in Phase II Clinical Trials. PMID- 17111562 TI - Weaning children from mechanical ventilation in a post-acute care setting. AB - As medical and technological advances have made it possible to prolong the life of children with chronic respiratory failure, children are being referred to post acute inpatient rehabilitation programmes. In these settings, children can be weaned from their ventilators and receive medical and rehabilitative care in a developmentally supportive environment at a lower financial cost than in an intensive care unit. There is strong evidence that weaning children from mechanical ventilation has beneficial effects on their functionality, ease of care and quality of life. There is, however, little scientific evidence describing how often successful weaning is achieved or the most effective methods. The purpose of this article is to present a consensus report detailing a structured approach to weaning children from mechanical ventilation in a post acute care setting. This study proposes a Weaning Severity Index and a Weaning Algorithm for use in the assessment and implementation of the weaning process in post-acute rehabilitation. Future clinical studies are needed to validate the suggested approach to ventilator weaning and to determine whether or not the weaning algorithm results in beneficial patient outcomes. PMID- 17111563 TI - Physician education on infant hearing loss in a developing country. AB - BACKGROUND: Physicians often account for delays in detection and intervention for infant hearing loss for reasons that are rarely investigated, especially in developing countries. This study, therefore, explores current physician knowledge, attitude and training towards infant hearing loss in Nigeria. METHODS: Knowledge of documented risk factors, intervention options and attitude towards timely intervention for infant hearing loss was investigated through a questionnaire survey among 246 final year medical students and 161 physicians drawn from tertiary hospitals in Lagos. RESULTS: Medical students were significantly less knowledgeable than practising physicians about most risk factors for hearing loss except for craniofacial anomalies, intra-uterine infections and low birthweight. Few respondents (11.4-42.9%) were knowledgeable about the risk of low birthweight and maternal exposure to noise in pregnancy for infant hearing loss. Medical students were significantly more aware of early detection possibility (67.8% vs 38.9 %, p=0.000) and more knowledgeable about the use of hearing aids for babies (57.3% vs 50.3%). Most respondents had a fairly positive attitude towards parental concern and early detection while medical students were significantly more positive in their attitude towards infant hearing screening (86.6% vs 74.5%, p=0.002). CONCLUSIONS: Medical students demonstrated greater awareness of early detection possibilities for infant hearing loss than physicians, although they had limited knowledge of some important risk factors. Continuing physician education reflecting evidence-based trends in child healthcare is necessary, while medical education also needs to be updated periodically. PMID- 17111564 TI - Evaluation of the nutrition counselling component of a fitness programme for children with disabilities. AB - OBJECTIVE: To describe and evaluate the nutrition counselling component of a fitness programme for children with disabilities. METHODS: Twenty-eight children with disabilities, ages 6-14 years, participated in a 16-week comprehensive fitness programme consisting of twice weekly exercise sessions, nutrition counselling and physical activity promotion education sessions. Nutrition sessions consisted of three individual and two group sessions. Individual nutrition goals were developed for each child using 3-day food intake diaries and parent interview. Body Mass Index (BMI) and progress towards nutrition goals were documented. RESULTS: No significant BMI changes were recorded for the entire group (n=28) or a sub-group with a goal to decrease BMI (n=8). Most of the children made improvements in individual goals indicating improvements in healthy eating habits. This included eating the daily recommended amount of servings of each food group, trying new foods and limiting foods containing saturated and trans fats, sodium and sugar. CONCLUSION: Short-term changes were noted in eating habits and behaviours during the 16-week fitness programme, although the effects did not influence overall BMI during the 16-week programme. Children with disabilities are at nutritional risk, and long-term follow-up is needed to determine if initial changes in parent-reported child eating behaviours will impact long-term nutrition, BMI and overall health. PMID- 17111565 TI - Comparing two types of augmentative and alternative communication systems for children with autism. AB - This study compared acquisition and preference for two types of augmentative and alternative communication (AAC) systems in three pre-schoolers with autism. Acquisition of requesting behaviour using a picture-exchange system vs a voice output communication aide (VOCA) was compared in an alternating treatments design. Following acquisition, both ACC systems were simultaneously available and the child could select which one of the two systems to use. There was little difference between picture-exchange and VOCA in terms of acquisition rates. Two children demonstrated a consistent preference for picture-exchange and the third showed a preference for the VOCA. Both speed of acquisition and system preference should be considered when designing AAC interventions for children with autism and related developmental disabilities. PMID- 17111566 TI - Fracture rate in children with cerebral palsy. AB - OBJECTIVES: To determine the prevalence of previous fracture, the rate of fracture over time and associated risk factors for fracture in children with moderate or severe cerebral palsy (CP). STUDY DESIGN: Three hundred and sixty four children with moderate-to-severe motor impairment (Gross Motor Function Classification System III, IV and V) enrolled in a multi-centre, region-based longitudinal study of growth, nutrition and health. Of these, 297 had baseline fracture information and 261 children had at least one follow-up assessment. Median duration of follow-up was 1.6 years, for over 600 person-years of follow up. RESULTS: Forty-six (15.5%) children reported 62 previous fractures at baseline assessment. Children with a history of fractures at baseline were older (mean age 11.9 vs. 8.9 years, p<0.0001) and had greater body fat (triceps z-score -0.01 vs. -0.68, p=0.0003) than children with no previous fracture. Twenty children (6.7%) reported 24 fractures during the follow-up period. Factors associated with risk of fracture during the follow-up period were higher body fat (p=0.03), gastrostomy use (p=0.05) and previous fracture (p=0.10). Based on 24 fractures in 604.5 person-years of follow-up, the rate of fracture was 4.0 per hundred children (4.0%) per year. For children with a history of fracture at baseline, the fracture rate was 7.0% per year; for children with gastrostomy, 6.8% per year; and for children with high triceps skinfold, 9.7% per year. CONCLUSIONS: Children with moderate or severe CP are at high risk for fracture. Children with greater body fat, feeding gastrostomy and prior history of fracture are at highest risk and may benefit most from intervention. Further longitudinal study and clinical trials in children with CP are needed to better understand the factors contributing to fracture risk in this population and the best methods of prevention and treatment. PMID- 17111567 TI - Using data visualization and signal processing to characterize the handwriting process. AB - INTRODUCTION: Disturbances in handwriting legibility and speed are found among elementary school-aged children. The aim of this paper is to present a set of sophisticated analytical tools suitable for visualization and evaluation of handwriting disturbances. METHODS: Handwriting samples from 30 children, 15 proficient and 15 non-proficient handwriters, aged 8-9 years were collected with the aid of a digitizing tablet. Temporal and spatial measures of the handwriting process dynamics based on signal processing methods were developed and visually presented. RESULTS: Significant differences between proficient and non-proficient handwriters were found in handwriting characteristics such as the standard deviations of letter width (t=2.96, p=0.008), letter height (t=3.24, p=0.005) and pen elevation (t=2.91, p=0.008). Significant differences were also found for the number of pen lifts (t=2.27, p=0.03), for the value of the correlation coefficients between letter length and time (t= -6.62, p=0.000) and between the actual and computed number of words (t=2.79, p=0.01). CONCLUSIONS: The techniques described in this paper provide objective measures for handwriting performance presented in a way designed to help clinicians and educators visualize handwriting difficulties during clinical evaluation and intervention. Data visualization and analysis appear to enhance information concerning the spatial and temporal dynamics of handwriting. PMID- 17111568 TI - Measuring fertility with the 1901 Canadian census: a critical assessment. PMID- 17111569 TI - [Is homogenization of milk allergy-inducing?]. PMID- 17111570 TI - [Port-a-cath insertion]. PMID- 17111571 TI - Incidence of neonatal tetanus in Kano State, Nigeria, 2006. PMID- 17111572 TI - Health conditions for travellers to Saudi Arabia for the pilgrimage to Mecca (Hajj). III. Poliomyelitis. Addendum. PMID- 17111573 TI - Performance of acute flaccid paralysis (AFP) surveillance and incidence of poliomyelitis, 2006 (data received in WHO headquarters as of 7 November 2006). PMID- 17111574 TI - "Let the record show": women and law in the United Counties of Durham and Northumberland, 1845-1895. PMID- 17111575 TI - Abrupt transition. PMID- 17111576 TI - Children in different and difficult times: the history of American childhood. Part 1. PMID- 17111577 TI - Ceramic microreactors for on-site hydrogen production from high temperature steam reforming of propane. AB - The steam reforming of hydrocarbon fuels is a promising method for the production of hydrogen for portable electrical power sources. A suitable reactor for this application, however, must be compatible with temperatures above 800 degrees C to avoid coking of the catalytic structures during the reforming process. Here, ceramic microreactors comprising high surface area, tailored macroporous SiC porous monoliths coated with ruthenium (Ru) catalyst and integrated within high density alumina reactor housings were used for the steam reforming of propane into hydrogen at temperatures between 800 and 1000 degrees C. We characterized these microreactors by studying C3H8 conversion, H2 selectivity, and product stream composition as a function of the total inlet flow rate, steam-to-carbon ratio (S/C), and temperature. As much as 18.2 sccm H2, or 3.3 x 104 sccm H2 per cm3 of monolith volume, was obtained from a 3.5 sccm entering stream of C3H8 at a S/C of 1.095 and temperatures greater than 900 degrees C. Operating at a S/C close to 1 reduces the energy required to heat excess steam to the reaction temperature and improves the overall thermal efficiency of the fuel processor. Kinetic analysis using a power law model showed reaction orders of 0.50 and -0.23 with respect to propane and steam, respectively, indicating that the rate limiting step in the steam reforming reaction is the dissociative adsorption of propane on the Ru catalyst. The performance of the microreactor was not affected after exposure to more than 15 thermal cycles at temperatures as high as 1000 degrees C, and no catalyst deactivation was observed after more than 120 h of continuous operation at 800 degrees C, making these ceramic microreactors promising for efficient on-site hydrogen production from hydrocarbons for use in polymer electrolyte membrane (PEM) fuel cells. PMID- 17111578 TI - Practical ethics. Diversity dilemma. PMID- 17111579 TI - Going 'lean' streamlines processes, empowers staff and enhances care. PMID- 17111580 TI - [What if Arab psychiatry preceded modern psychiatry?]. PMID- 17111581 TI - U.S. health care system rates 66 out of 100 on scorecard assessing efficiency, quality, outcomes, equity, access. PMID- 17111582 TI - Broader use of quality measures in Medicare pay-for-performance project could save lives, money. PMID- 17111583 TI - Who decides: do individual physicians in group practices have discretion over acceptance of new Medicaid patients? PMID- 17111584 TI - Description and implementation of a quality control program in an imaging-based clinical trial. AB - RATIONALE AND OBJECTIVES: The American College of Radiology Imaging Network is participating in the National Lung Screening Trial, a large, multicenter, randomized controlled trial, comparing multidetector helical computed tomography (MDCT) versus chest radiography (CXR) in screening for lung cancer. Because the threshold for detection of disease is an inherent function of image quality, and consistent image quality is necessary to track changes in suspicious findings, our purpose was to develop an image quality control (QC) program across all clinical sites for both modalities. MATERIALS AND METHODS: The primary goals of the QC program include standardization of imaging protocols, certification of imaging equipment, and ongoing, periodic evaluation of the equipment calibration and image quality. Minimum standards for equipment and standardized cross platform acquisition protocols are achieved via radiologist and physicist attestation forms and web-distributed technique charts, respectively. Imaging equipment performance standards are implemented through an initial machine certification process that includes equipment calibration. Ongoing assessment of equipment performance and calibration, as well as adherence to established imaging protocols. is accomplished via periodic submission of calibration records and phantom images. Participant-specific image acquisition parameters are entered into a web-based centralized database and variations from established protocols are automatically flagged for review. Participant radiation dose can be estimated from the image acquisition parameters applied to the imaging equipment calibration measurements. A radiologist visual review committee also evaluates participant images for diagnostic quality. Data are collected from 23 independent centers, representing 14 models of MDCT scanners from four manufacturers, and CXR systems that include film-screen, computed radiography, and direct digital radiography systems. RESULTS: Widespread imaging protocol variation in extant clinical practice-as well as variability in equipment technology, image acquisition parameters, manufacturer terminology, and user interface-have required careful standardization as a prerequisite to trial participation and ongoing image QC. Acceptable ranges for image acquisition parameters have been refined to accommodate continuously evolving equipment platforms and the scope of participant size and body habitus. CONCLUSION: Standardization of imaging protocols is a critical component of image-based clinical trials, predicated on ongoing dialogue between sites and a centralized review committee. PMID- 17111585 TI - [The "sick black" in America, 1850-1900]. PMID- 17111586 TI - [Vladimir Iakovlevich Aleksandrov (the 100th anniversary of his birth)]. PMID- 17111587 TI - Dental postcards No. XXXIII. PMID- 17111588 TI - 19th century dentistry advertising trade cards. PMID- 17111589 TI - Dental postcards No. XXXIV. PMID- 17111590 TI - [Japan, research, education, and physiology]. PMID- 17111591 TI - Stress and depression in the employed population. AB - OBJECTIVES: This article describes stress levels among the employed population aged 18 to 75 and examines associations between stress and depression. DATA SOURCES: Data are from the 2002 Canadian Community Health Survey: Mental Health and Well-being and the longitudinal component of the 1994/95 through 2002/03 National Population Health Survey. ANALYTICAL TECHNIQUES: Stress levels were calculated by sex, age and employment characteristics. Multivariate analyses were used to examine associations between stress and depression in 2002, and between stress and incident depression over a two-year period, while controlling for age, employment characteristics, and factors originating outside the workplace. MAIN RESULTS: In 2002, women reported higher levels of job strain and general day-to day stress. When the various sources of stress were considered simultaneously, along with other possible confounders, for both sexes, high levels of general day to-day stress and low levels of co-worker support were associated with higher odds of depression, as was high job strain for men. Over a two-year period, men with high strain jobs and women with high personal stress and low co-worker support had elevated odds of incident depression. PMID- 17111592 TI - Unhappy on the job. PMID- 17111593 TI - Government-subsidized home care. PMID- 17111594 TI - Seniors' use of home care. PMID- 17111595 TI - [Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (2004). III. Secular changes in susceptibility]. AB - The bacteria (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella spp. and Pseudomonas aeruginosa) isolated from patients diagnosed as urinary tract infections (UTIs) in 14 institutions in Japan were collected between August 2004 and July 2005. The susceptibilities of these bacteria to various antimicrobial agents were measured. The bacteria were divided into 2 groups consisting of uncomplicated UTIs and complicated UTIs (with and without indwelling catheter) based on their isolation origins. The results were compared with those obtained between 1995 and 2003. The drug sensitivity of S. aureus in this year was similar to those in up to the previous year and S. aureus showed the best susceptibility to vancomycin (VCM) and arbekacin (ABK). The drug sensitivity of E. faecalis in this year also was similar to those in up to the previous year. The susceptibility of E. coli to cephems in this year was generally good and was similar to those in up to the previous year. MIC90 of cefozopran (CZOP) was the most stable and 0.125 microg/mL or less since 1995. The susceptibility of E. coli to cefpirome (CPR) and cefotiam (CTM) also was good but to cefaclor (CCL), cefixime (CFIX), and cefpodoxime (CPDX) was largely decreased in complicated UTI groups. The sensitivity of E. coli to carbapenems also was good but to carumonam (CRMN) tended to decrease. The susceptibility of E. coli to quinolones, however, has largely changed and has decreased since 2003 in uncomplicated UTIs and 2000 in complicated UTIs. That was suggested the development of the resistance to the drug. The susceptibility of Klebsiella spp. to cefazolin (CEZ), CTM, CCL, CPDX, and cefditoren (CDTR) decreased in the previous year and recovered to the year before the previous year in this year. The susceptibility of Klebsiella spp. to other cephems was stable since 1995, especially against CZOP, the highest sensitivity (MIC90: < or = 0.125 microg/mL) was maintained. The susceptibility of Klebsiella spp. to carbapenems and CRMN also was good. The susceptibility of Klebsiella spp. to aminoglycosides was lower than to CZOP but was stable since 1995. The susceptibility of P. aeruginosa was generally low and has largely changed against the majority of the agents since 1995. The susceptibility of P. aeruginosa isolated from uncomplicated UTIs has largely changed against ceftazidime (CAZ), cefsulodin (CFS), CZOP, imipenem (IPM), meropenem (MEPM), aztreonam (AZT), CRMN, gentamicin (GM), and tobramycin (TOB). The susceptibility of P. aeruginosa isolated from complicated UTIs has largely changed against CSF, CZOP, MEPM, GM, and ciprofloxacin (CPFX). The susceptibility of P. aeruginosa isolated from complicated UTIs has been stable against amikacin (AMK). For annual changes in MIC50, TOB and IPM had a relatively stable and high activity (MIC50: 0.5-2 microg/mL). PMID- 17111596 TI - [Antibacterial activity of nadifloxacin against Staphylococcus and Propionibacterium isolated from patients with dermatological infections]. AB - Nadifloxacin (NDFX) is a fluoroquinolone antibiotic developed by Otsuka Pharmaceutical Co., Ltd. and is used as a topical drug for the treatment of infections in the field of dermatology. We investigated the susceptibility of a total of 575 strains (two kinds of Staphylococcus species and Propionibacterium species which were isolated from patients with dermatological infections for 3 periods, i.e., 1996, 2000 and 2005) to NDFX and other reference antibiotics. The minimum inhibitory concentration of the four antibiotics, NDFX, levofloxacin (LVFX), clindamycin (CLDM) and gentamicin (GM), against the test strains were determined by the agar dilution methods, in according with the Japan Society of Chemotherapy. The antibacterial activity of NDFX against methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA), Staphylococcus epidermidis and P. acnes was the most potent of all the antibiotics tested, and there were no test organisms which became resistant to NDFX with period. The MIC90 values of NDFX for the four test organisms isolated in 2005 were 0.05 microg/mL; MSSA, 1.56 microg/mL; MRSA, 0.78 microg/mL; S. epidermidis and 0.20 microg/mL; P. acnes, respectively. On the other hand, there were LVFX-, CLDM- and GM-resistant MRSA. The MIC50 values of CLDM and GM for MRSA were >100 and 25 microg/mL, respectively. The MIC50 value of GM for P acnes was 12.5 microg/mL, but NDFX was potently active against these organisms as compared with these two antibiotics and the MIC50 values of NDFX were 0.05 microg/mL for MRSA and 0.20 microg/mL for P. acnes. These results suggest that NDFX is even at present useful as an antibiotic for the treatment of infections in the field of dermatology though it is more than 12 years since the approval to manufacture and sell the drug was obtained in 1993. PMID- 17111597 TI - [VOC emission situation and control measures of gas station in China]. AB - The emission factor is used to estimate the volatile organic compound (VOC) emission caused by gas station. After considering the economical, social and population factors, the activity rate was modified, and then the fuel consumption and VOC emission trend in the next 20 years can be predicted. The result shows the VOC emission from gas station in China 2002 was 187.6kt and this number will increase to 1196kt in 2030 if no further control measures will be implemented. And the economic loss caused by gasoline vapor arrived to 0.75 billion RMB in 2002 and will be 4.78 billion RMB in 2030. The cost-benefit approach of the commercially available gasoline vapor recovery technologies in China included Stage I, Stage II and on-board refueling vapor recovery (ORVR) were analyzed, and the result shows introducing these three systems will bring larger reduce of VOC emission, and the combination of them can do a even better job. Compared with Stage II, ORVR is more efficient and cheaper, but it will take long time to implement ORVR. And it will take at least 11 years in China to convert to ORVR above 80%. So Stage II vapor recovery system may be a short term option while ORVR should be treated as the ultimate solution for controlling the vapor emission from gas stations in the future. PMID- 17111598 TI - [Measurement of the CO2 flux on the water-air interface of Taihu Lake]. AB - Based on the measurement of the CO2 flux on the water-air interface of Taihu Lake by closed chamber technique from January 2003 to June 2005, variation characteristics of the CO2 flux on the water-air interface in Taihu Lake are analyzed. Diurnal variation of the CO2 flux on the water-air interface is obvious. The CO2 flux is -0.79 mg/(m2 x h) in spring, -4.89 mg/(m2 x h) in summer, -4.06 mg/(m2 x h) in autumn and -2.56 mg/(m2 x h) in winter. Taihu Lake is the sink of carbon monoxide. The CO2 flux gets larger in severer polluted area of the lake. Seasonal variation is not apparent in the algae type lake, but obvious in grass type lake. It is high in summer and autumn, and low in spring and winter. Correlative factors of CO2 flux on the water-air interface of Taihu Lake possibly include weather, solar radiation, wind velocity, temperature, pH, TA, Chla, TC, TN and TP. PMID- 17111599 TI - [Mercury exchange fluxes between air and soil interface over different type of land in Wanshan Hg mine area]. AB - Air/soil Hg exchange fluxes were measured using field chamber-automated air mercury analyzer method over different land in Wanshan Hg mine area in two seasons. The results showed that the air/soil Hg exchange fluxes were very strong. The highest Hg emission flux from soil was 27 827 ng/(m2 x h), the highest Hg deposition flux from the atmosphere was 9 434 ng/(m2 x h). Because of Hg emission from anthropogenic activities and natural lands, the Hg concentrations in air in Wanshan Hg mine area are 1 - 3 orders of magnitude greater than background area. The highest average Hg concentration in air reached 1 101.8 ng/m3, and the lowest average Hg concentration in air still reached 17.8 ng/m3. These indicated that the atmosphere was polluted seriously in Wanshan Hg mine area. The Hg exchange fluxes are influenced by solar irradiation and the Hg concentrations in air. The solar irradiation accelerates the Hg emission from soil. Conversely, the Hg concentration in air restrained the Hg emission from soil, and even leads the Hg concentration depositing to soil surface. The Hg emission fluxes from uncovered soil are higher than that from covered soil by vegetations significantly. And the slag becomes net atmospheric Hg source. PMID- 17111600 TI - [Stable carbon isotope analysis method for the atmospheric formaldehyde]. AB - A method of compound-specific stable carbon isotopic analysis for the research about the sources of atmospheric formaldehyde was preliminary studied using gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) via 2, 4 dinitrophenylhydrazine (DNPH) derivatization. In order to evaluate the reproducibility, the accuracy and the carbon isotope effects of the method, formaldehyde with different delta 13C values were used to simulate the sampling procedure. The results show that the maximal analytical deviation for all formaldehyde 2, 4-dinitrophenylhydrazone is 0.3 per thousand and the average deviations between the determined and theoretical delta 13C values of them are 0.24 per thousand +/- 0.14per thousand (ranged from 0.03 per thousand to 0.35 per thousand), less than 0.5 per thousand the technical specifications of the GC/C/IRMS system. These mean that no carbon isotopic fractionation occurred during the procedure. The study for the indoor and outdoor atmospheric formaldehyde in the restaurant show that the stable carbon isotopic compositions are significant different for different sources of formaldehyde. The present method could provide valuable information about the sources of atmospheric formaldehyde. PMID- 17111601 TI - [Photocatalytic degradation of hexafluorobenzene by tungstophosphoric acid]. AB - We verified that C6F6, a typical perfluorine compound, could be photocatalytic degraded by H3PW12O40. The defluorination of C6F6 was determined as a function of irradiation time, pH, initial concentration of H3PW12O40, and additives as well. The defluorination of C6F6 increased with the irradiation time. Acid condition, especially pH = 1.0 is preferred. 3 mL solution of 2.0 x 10(-4) mol/L C6F6 and 5.0 x 10(-6) mol/L H3PW12O40 was irradiated for 20 min and the defluorination of C6F6 was equal to 208.1%. O2, KMnO4 and K2S2O8 added in the solution were able to increase the defluorination of C6F6 via oxidating [PW12O40](4-). The defluorination of C6F6 would decline in the presence of alcohol and aromatic compounds. PMID- 17111602 TI - [Catalytic removal of NO on modified precious catalyst]. AB - The streams of waste gases containing NO from industries would cause seriously pollution on the environment if they are directly discharged without further purification. The property of removal of NO from the waste gas at lower temperature performed on platinum catalyst by reduction with NH3 was investigated. Experimental study showed that the precious catalyst, platinum, had good activity at low temperature and high selectivity of catalytic removal of NO from the waste gas by ammonia. However, its performance was affected by the sulfur compound, SO2, which was present in the waste gas. Results indicated that the performance of platinum supported catalyst could be improved with addition of lanthanides on it. The suitable compositions of the modified catalyst were 1:3.78:3.56 molar ratio of Pt:La:Ce in the experiment. PMID- 17111603 TI - [Fluorine removal efficiency of organic-calcium during coal combustion]. AB - Effectiveness of calcium magnesium acetate (CMA) and calcium acetate(CA) as feasible HF capture were studied by means of fixed bed tube furnaces. The effects of temperature, particle diameter and Ca/S molar ratio on the fluorine removal efficiency were studied. By contract with CaCO3 at the same condition, we find that the HF capture effectiveness of those sorbents is superior to CaCO3, especially at high temperature. At 1 000 - 1 100 degrees C, the efficiency of fluorine removal during coal combustion of CMA is 1.68 - 1.74 times as that of CaCO3; the efficiency of fluorine removal during coal combustion of CA is 1.28 - 1.37 times as that of CaCO3. PMID- 17111604 TI - [Semi-analysis algorithm to retrieve pigment concentrations in the red tide area of the East China Sea]. AB - For the purpose of detecting and forecasting research of red tides to reduce the loss, a semi-analytic algorithm to retrieve chlorophyll-a concentrations was established in the area where red tides often brought out, according to the data collected during the red tides cruise in the East China Sea in April 2002. In the algorithm, empirical equations were made based on the coefficients from the in situ data, including the optical properties of the research area. The in-situ data were used to validate the algorithm. The discrepancy of chlorophyll-a absorption coefficients and concentrations are mainly located in the region of 30%. The root mean deviation of the chlorophyll-a concentrations between the observed and the calculated is 0.24, the maximum relative deviation 40.93%, the mean relative deviation 18.83% and the correlation coefficient 0.83. The results show that the precision of the algorithm is high and the algorithm is fit for the research area. PMID- 17111605 TI - [Mechanisms of removing red tide organisms by organo-clays]. AB - We tested the influence of the preparation conditions of the quaternary ammonium compounds (QACs) modified clays on their capacities to remove red tide organisms, then discussed the mechanisms of the organo-clays removing red tide organisms. Hexadecyltrimethylammonium (HDTMA) improved the capacity of clays to flocculate red tide algae, and the HDTMA in metastable state enhanced the toxicity of the clay complexes to algae. The capacities of the organo-clays correlated with the toxicity and the adsorbed amount of the QACs used in clays modification, but as the incubation time was prolonged the stability of the organo-clays was improved and the algal removal efficiencies of the clay complexes decreased. When the adsorbed HDTMA was arranged in different clays in which the spatial resistance was different, there was more HDTMA in metastable state in the three-layer montmorillonite. Because of the homo-ion effect the bivalent or trivalent metal ions induced more HDTMA in metastable state and the corresponding organo-clays had high capacities to remove red tide organisms. When the reaction temperature was 60 degrees C the adsorbed HDTMA was easily arranged on cation exchange sites, if the temperature rose or fell the metastable HDTMA would increase so that the capacity of the clays was improved. PMID- 17111606 TI - [Evaluating comprehensive quality of sediment in Dianchi Lake using adjusted AHP method and 137Cs dating]. AB - It is difficult to evaluate comprehensive quality of sediment and to understand development trend of pollution because of absence of monitoring data, especially history data. Combining the method of 137Cs dating with the ways of general sampling and measurement can easily resolve the problem of absence of data and also provide the possibility for calculating weighted environmental quality comprehensive index using the adjusted analytical hierarchy process (AHP) method. In order to overcome the willfulness the judgment matrix is formed objectively based on calculating monitoring data. Based on the monitoring data of sediment pollution and the weights of various factors gained by adjusted AHP method the comprehensive quality of sediment in each zone of Dianchi Lake was evaluated and the results indicated that the pollution of sediments in each zone at the present be serious more than that in the history. The condition may be related to the industrial development and distribution of industries in Dianchi Lake basin. Therefore, in order to improve the comprehensive quality of sediment in Dianchi Lake and to prevent the secondary pollution of heavy metals in sediment from happening, it is necessary to control the pollutants discharge and to remove the pollutants with various ways. PMID- 17111607 TI - [Space-time dependent variances of ammonia and phosphorus flux on sediment-water interface in Lake Taihu]. AB - Monthly research of ammonia and phosphorus fluxes in water-sediment interface in East Lake Taihu (ELT, macrophyte dominated) and Meiliang Bay (MB, algae dominated) was processed with intact sediment cores' incubation and pore water diffusive model. The ammonia and phosphorus fluxes calculated with intact sediment cores' incubation (F(i)) showed discrepancy in different lake zones with different ecotype. The yearly average fluxes of ammonia and dissolved phosphorus in ELT were (44.9 +/- 21.9) mg x (m2 x d)(-1) (Mean +/- SD) and (2.06 +/- 1.71) mg x (m2 x d)(-1); and (16.2 +/- 12.0) mg x (m2 x d)(-1) and (0.53 +/- 0.52) mg x (m2 x d)(-1) in MB. The molecular diffusive fluxes (F(m)) of the two lake zones showed the same pattern, but the absolute value difference may as high as an order of magnitudes. So this molecular diffusive model cannot be used to evaluate the nutrients fluxes in sediment-water interface in Lake Taihu, a lake vulnerable with wind and wave affection and benthic bio-disturbation. Compared with the two lake zones with different ecotypes, macrophyte dominated one had higher nutrients fluxes than the algae one. ELT had higher bio-disturbation, which was revealed by the F(i)/F(m) ratio. When the dissolved oxygen (DO) remained in high concentration, known as aerobic condition, the nutrients' fluxes were not correlated with the DO. Similarly, the fluxes were not correlated with the nutrients' concentration of the overlying water at the existing condition. The discrepancy between higher nutrients' fluxes and lower nutrients loading in ELT suggested the higher particulate settlement rate and assimilate rate promoted by the macrophyte. This is the important theoretical basis of rebuilding the healthy ecosystems with restoring the water plants. PMID- 17111608 TI - [Scenario analysis of integrated model of nutrients in the Miyun Reservoir and its watershed]. AB - Supported by the integrated model of nutrients for the Miyun Reservoir in part I, effects of different control measures were studied on the water quality of the reservoir. Four scenarios were assumed and analyzed. Results of the base case scenario showed that TN concentration of the Miyun Reservoir had highly exceeded the environmental quality standard for surface water, and TP was relatively better. Furthermore, there were many regions that chlorophyll-a concentration exceeded 10 microg/L in the reservoir, and centralized in the reservoir area of Chaohe River. Scenario 1, 2 and 3 investigated effects of different pollution control measures on the water quality of the reservoir. Results showed that the control of nutrient input loads could improve the water quality greatly, especially control of TP loadings would limit algae growth effectively, and regions that chlorophyll-a concentration exceeded 10 microg/L even disappeared. The results indicated that some control measures, such as changing farming style, part treatment on stockbreeding pollution and reducing point source pollutant loadings were very effective and essential to decrease the eutrophic level of the reservoir. PMID- 17111609 TI - [Substance flow analysis on phosphorus cycle in Dianchi basin, China]. AB - Investigation of physical profiles of nutrients within socio-economic systems is one of the key approaches to cure eutrophication. Applying a substance flow analysis (SFA) method for the case of Dianchi basin located in southwest of China, we established a regional static SFA model (PHOSFAD) in 2000 via balancing societal phosphorus flows. Aggregate characteristics of the overall phosphorus throughput, and physical efficiencies in relation to phosphorus uses of production and consumption sectors, including mining, processing, farming, livestock husbandry and household consumption, were subsequently identified. The methodology and results of this study illustrate applicability and value for rational decision-making aiming to curb eutrophication of Dianchi Lake. PMID- 17111610 TI - [Synergistic effects by P and N pollution to fluctuation behavior/bloom of algae along the Three-Gorges valley]. AB - Absorption rate coefficient of algae omega(i) to nutrients such as N and P could be used for describing algal increases/decreases velocity in water areas in theory. omega(i) raise might correspond to algal quickly growth and to ccelerate absorption of N and P while omega(i) decrease might correspond to algal decompose and release of N and P. According to locale measuring data along the Three-Gorges valley and algal dynamics model of nutritious absorption we have obtained some interest 3-dimension figures in which omega(i) will varies up and down obviously with N and P concentration in special bound to show a synergistic effects of N and P that might reveal an inner behavior of algal bloom/decompose. The research results explain in reason: (1) algal blooms do will happen in one special P/N range in a certain water system; (2) when omega(1) and omega(2) ascend rapidly and simultaneously in positive direction at same time algae would bloom, and when omega(1) and omega(2) descend sharply and simultaneously in negative direction at same time algae would decompose; (3) The velocity of algal bloom is not only same approximately as one of algal decompose, but also its variety has evidently periodic fluctuation. All of these could reveal effectively mechanism of nutritious absorption/release as algal bloom/decompose. PMID- 17111611 TI - [Treatment performance and enhancement of subsurface constructed wetland treating polluted river water in winter]. AB - Treatment performance of subsurface constructed wetland treating polluted river water in winter was studied. When hydraulic loading fell down from 30cm/d to 15cm/d, the average removal rates of ammonium nitrogen and COD rose up from 14% and 20% to 39% and 31% respectively. Water temperature had great influence on ammonium nitrogen removal. Membrane overlay on the wetland could improve pollutants removal efficiently. The average removal rates of ammonium nitrogen and COD could rise up from 29.4% and 29% to 67.6% and 46.6% respectively. Microorganism enzyme activity experiments show that increase of microorganism activity caused by membrane overlay contributed a lot to pollutants removal enhancement. Urease activity rose up from 0.025 mg/(g x d) to 0.037 mg/(g x d) with membrane overlay, and dehydrogenase activity rose up from 0.17 microL/(d x g) to 4.54 microL/(d x g) with membrane overlay. The pollutants releasing experiment of aquatic plants show that faded plants would release a lot of pollutants into water. Therefore, plants harvest was necessary in autumn. PMID- 17111612 TI - [Study on the characteristics of combined sewer overflow from the high density residential area in Shanghai]. AB - The urban non-point pollution has become main pollution resource of urban water bodies of Shanghai. Character of combined sewer overflow from watershed SA in Shanghai was studied, and the correlation of influence factors to the EMCs of overflows was discussed. It is found that the EMCs of COD, BOD, SS, NH3-N, TN, TP are 614 mg/L, 208.5 mg/L, 684 mg/L, 17.6 mg/L, 29.8 mg/L, 3.0 mg/L respectively, and the values obtained herein are much higher than the documented data from other countries. From the probability plot of the EMCs, it is found that the BOD has the best fitness for lognormal distribution; and correlation between the EMCs of COD, SS and the ratio of antecedent dry weather time to rain duration is quite good. PMID- 17111613 TI - [Anaerobic degradation kinetics of 2,4-dinitrophenol]. AB - With glucose as co-substrate, anaerobic degradation kinetics of 2,4-dinitrophenol (2,4-DNP) were investigated in batch culture. The results show that 2,4-DNP and glucose can be degraded by the bacteria simultaneously. Although the effect of COD on 2,4-DNP degradation was minimal, addition of 2,4-DNP effected COD degradation obviously. The rate of 2,4-DNP degradation increased with increasing initial 2,4-DNP concentrations up to 225 mg/L. Further increase in initial 2,4 DNP concentrations caused decrease in the rate of degradation because of substrate inhibition. Uncompetitive inhibition equation is proposed to describe the degradation of 2,4-DNP. With non-linear regression technology, the kinetic model parameter q(max), K(s) and K(i) are found to be 3.24 mg/(h x g), 196.23 mg/L and 165.91 mg/L respectively. The experimental data verification for the model equation is satisfactory. PMID- 17111614 TI - [Effect of nitrite on aerobic phosphate uptake by phosphate accumulating organisms]. AB - Effect of nitrite at various concentration levels on aerobic phosphate uptake was investigated through a series of batch experiments. Furthermore, the effect of nitrite accumulated in the process of nitrification on aerobic phosphate uptake was studied in saline wastewater treatment. The results show that NO2(-) -N concentration of 4 mg/L inhibits aerobic phosphate uptake by phosphate uptake by phosphate accumulating organisms (PAO) and phosphate uptake rate decreases 8%. Exposure to higher nitrite concentration levels inhibits aerobic phosphate uptake severely. At NO2(-) -N concentration of 15 mg/L, phosphate uptake rate decreases 61%. The toxic effect of nitrite is presumed to be linked with free nitrous acid (FNA). Significant inhibition on aerobic phosphate uptake appeared at 0.000 2 mg/L of FNA concentration. The inhibiting effect of nitrite is found to occur only when nitrite is present. The ability of accumulating phosphate resumes afternitrite is no longer present. During the process of nitrification of nitrification of saline wastewater, the inhibition of nitrite on aerobic phosphate uptake is slight at the initial 1 - 2 h due to low nitrite accumulation. With the build-up of NO2(-) -N(up to about 8 to 9 mg/L), the inhibiting effect of nitrite increases gradually. It is found that higher ammonium concentration causes lower pH value and higher FNA concentration, which could decrease the amount of phosphate uptake. PMID- 17111615 TI - [Impact of catalytic ozonation with ferric hydroxide on HAAs formation potential of a filtered surface water]. AB - Formation potential of five haloacetic acids (HAA5FP) of a filtered surface water was studied after ozonation alone and catalytic ozonation with a ferric hydroxide (FeOOH). Factors studied were oxidation time, bromide, pH, bicarbonate alkalinity, and ozone dosages. The haloacetic acids detected were dichloroacetic acid (DCAA), trichloroacetic acid (TCAA), and dibromoacetic acid (DBAA) when bromide existed. The catalytic ozonation caused a reduction of HAA5FP of the non bromide containing water for 9.5% - 18.3% compared to that of ozonation in 5-20 minutes. Incremental addition of bromide led to a much lower HAA5FP after catalytic ozonation than that after ozonation. HAA5FP of the water at neutral pH was higher than that at acidic and basic conditions. Catalytic ozonation showed a most advantageous potential in controlling HAA5FP over ozonation at neutral pH. The HAA5FP decreased as bicarbonate concentration increased, and the disparity between ozonation and catalytic ozonation was also reduced. The HAA5FP after catalytic ozonation was 11.2% to 28.0% lower than that after ozonation while the ratio of O3/TOC ranging from 0.45 to 1.43. The effect of catalytic ozonation on HAA5FP of the water is closely related to its enhanced generation of hydroxyl radicals in catalytic process. PMID- 17111616 TI - [Electrochemical hydrodehalogenation of pentachloraphenol in aqueous solution by porous titanium loaded Pd cathode]. AB - A study on electrochemical hydrodehalogenation of pentachloraphenol in aqueous solutions was conducted on porous titanium loaded Pd cathode and the operational parameters were investigated. Chloride ions and phenol is the main products analyzed by GC-MS. Up to 100% electrochemical hydrodehalogenation can be achieved with more than 90% conversion to phenol. The result showed that current in the circle play the most important role of dehalogenation while the best parameter is 300mA in this study. Compared to current, the effect of pH value and flow rate on the current efficiency and energy consumption of dehalogenation is limited. Complete dehalogenation, high current efficiency, low energy consumption and operational convenience confirm the feasibility of this method. PMID- 17111617 TI - [Kinetics of Electro-Fenton treatment of leachate from municipal solid waste incinerator]. AB - SPR electrode was used in Electrio-Fenton treatment on the effluent from biological and coagulative treatments of solid waste incinerator leachate, at the same time, the kinetics model for the COD effluent concentration were set up. The result indicated that the disposal curve of COD confirm to the pseudo third order kinetics, and the formula is c(t) = (2kt + c(0)(-2))(-0.5), in which the c(t) is the COD concentration of treatment effluent. With the statistical analysis to the disposal curve of COD from Electro-Fenton treatment in different conditions, the experience mathematics model of k value was set up, in which the current density, pH value and COD concentration of influent are effective factors. Moreover, with the comparison between the simulative COD of effluent and the experimental results, it was showed that this kinetic model could be used to investigate the COD disposal efficiency of Electro-Fenton treatment on the effluent from biological and coagulative treatments of solid waste incinerator leachate preferably. PMID- 17111618 TI - [Treatment of domestic wastewater using sequence batch reactor enhanced by low intensity ultrasound]. AB - Enhancement effect of ultrasound (US) on the bio-treatment of domestic wastewater was studied through contrast experiments between two sequence batch reactors (SBR) with and without US. The results showed that when US with intensity of 0.3 W x cm(-2) was employed to irradiate 10% of the activated sludge in the SBR for 10 minutes every 8 hours, the removal rate of COD was improved by 3% - 6%, and the effluent COD of US-SBR was lower than that of the control SBR by 40% - 53%. Moreover, the tolerance of SBR to high concentration and toxicity impact was improved effectively. For the practical domestic wastewater, the sludge activity was 14% higher in the US-SBR than that in the control SBR, increasing the capacity of microorganism to degrade the refractory compounds. The settleability of activated sludge in the US-SBR descended slightly and 5% increase of SVI (sludge volume index) was observed during the experiment, but it had little influence on the system since the increment was very small. According to the results of scan electro-microscope (SEM) and microbial community structures analysis, the microorganism in the US-SBR was damaged by US and compared with the control, the cell wall was ruffled obviously, but it didn't change the community structures of activated sludge. Kinetics analysis of microbial respiration in the two SBRs showed that the activated sludge in the US-assisted SBR could make better use of substrate. PMID- 17111619 TI - [DBPs formation characteristics in chlorinating and relationship with chemical structure of some aromatic organic matter]. AB - Some organic matter that have aromatic structure similar as humic acid are selected to conduct chlorination test, determined the formation of HAAs and THMs of tested matters, researched the relationship of the chemical structure of DBPFP with DBPs formation characteristics. The results indicated that: (1) The sequence of HAAs formation activity is p-Hydroxybenzoic acid > p-Aminoacetophenone > o Benzenediol > m-Benzenediol; and THMs' s is m-Benzenediol > p-Aminoacetophenone > 2-Hydroxy-p-aminoacetophenone > p-Hydroxybenzoic acid. (2) The sequence of earlier chlorination reaction rate of HAAs is m-Benzenediol > p-Aminoacetophenone > p-Hydroxybenzoic acid > o-Benzenediol; and THMs' s is p-Hydroxybenzoic acid > 2 Hydroxy-p-aminoacetophenone > m-Benzenediol > p-Aminoacetophenone. (3) The substances that having the substituent group structure of para and ortho position are mostly format HAAs, and one's of having meta position are mostly format THMs. (4) On the benzene ring, hydroxyl group has the highest activity of forming DBPs, and the second is ketone group; amino-group is play a part of auxiliary forming para, ortho or meta position structure mainly; aldehyde and carboxyl group are inert function group; the function of methyl group is remain to be determined. PMID- 17111620 TI - [Effects of terminal products on hydrogen production by ethanol hydrogen producing microflora]. AB - The effects of terminal products on hydrogen production were conducted in continuously stirred tank reactor (CSTR) by the acclimated ethanol hydrogen producing bacteria. Glucose was added as carbon source and different ambient pH was carried out by buffer solution. High ethanol production was simultaneously achieved with high hydrogen production. The addition of 40 mol/L ethanol or acetic acid can result in the reduction of hydrogen production, the former was 34% and the latter was 84.3%. The ethanol fermentation formation for mixed microflora was attributed to the natural selection. PMID- 17111621 TI - [DNA extraction methods of compost for molecular ecology analysis]. AB - Molecular ecology provides new techniques for studying compost microbes, and the DNA extraction is the basis of molecular techniques. Because of the contamination of humic acids, it turns to be more difficult for compost microbial DNA extraction. Three different approaches, named as lysozyme lysis, ultrasonic lysis and proteinase K lysis with CTAB, were used to extract the total DNA from compost. The detection performed on a nucleic acids and protein analyzer showed that all the three approaches produced high DNA yields. The agarose gel electrophoresis showed that the DNA fragments extracted from compost had a length of about 23 kb. A eubacterial 16S rRNA gene targeted primer pair (27F and 1 495R) was used for PCR amplification, and all the samples got almost the full length 16S rDNA sequence (about 1.5 kb). After digested by restriction endonucleases (Hae Ill and Alu I), the restriction map showed relatively identical microbial diversity in the DNA, which was extracted by the three different approaches. All the compost microbial DNA extracted by the three different approaches could be used for molecular ecological study, and researchers should choose the right approach for extracting microbial DNA from compost based on the facts. PMID- 17111622 TI - [Isolation and characterization of a facultative anaerobic aniline-degrading bacterium]. AB - An aniline-degrading bacterium (designated strain AN29) was isolated from dyeing wastewater process (anaerobic baffled reactor, ABR) with the capability of utilizing aniline as sole carbon source and nitrogen source. It was identified as Pseudomonas sp. based upon the phenotypic properties and a partial analysis of the 16S rDNA. The strain could degrade aniline under the aerobic and anaerobic conditions, the optimal initial pH 6.5 - 8.0, a temperature of 37 degrees C, and initial aniline concentrations of 500 - 2 000 mg/L with maximum concentration of 4 000 mg/L respectively. PMID- 17111623 TI - [Effect of trace element on the growth of white rot fungus and suppressing yeast in liquid medium]. AB - Growth of white rot fungus in liquid medium with different trace element and effect of these medium on suppressing the growth of yeast under non-sterile were investigated in agitated Erlenmeyer flasks. Results showed that the size and amount of mycelium pellets in liquid medium with trace element and Fe 3.5 mg/L was more than that with trace element but no iron and without any trace element under sterile condition; moreover, the size and amount of mycelium pellets increased with the rise of the concentration of trace element and iron. In addition, if applying the mode of incubating under sterile condition and degrading under non-sterile condition, the decolorization of reactive brilliant red K-2BP in liquid medium without any trace element and with trace element but no iron was similar to that of degrading under sterile condition; but the decolorization in liquid medium with trace element and Fe 3.5 mg/L was less than the results under sterile condition. Through observing under microscope, it was found that the amount of yeast in liquid medium with Fe 3.5 mg/L was more than the medium without any trace element and with trace element but no iron. Therefore, trace element, especially iron, plays a very important role in suppressing yeast in liquid medium; furthermore, iron also is necessary to the growth of whit rot fungus. So that, to control the concentration of iron in liquid medium could reduce the invasion of yeast under non-sterile condition as long as it does influence the growth of white rot fungus. PMID- 17111624 TI - [Detecting organophosphorus pesticide in water environment using an enzyme biosensor]. AB - Based on a screen printed carbon electrode which contained a sensitive film prepared by crosslinking acetylcholine esterase (AChE) with glutaraldehyde on the surface of working electrode, an amperometric enzyme biosensor was taken on the determination of organophosphorus (OP) pesticide in water environment. Cyclic voltammetry was applied to assess the coherence of prepared screen printed electrodes. The results show that the variation is less than 10%. The sensor sensitivity was optimized with respect to content of enzyme, crosslinker and substrate. The best sensitivity is obtained using 0.2% of glutaraldehyde, 0.5 mg/mL of AChE and 10 mmol/L of acetylthiocholine chloride (ATChCl). Amperometric I-t curve was conducted to detect the decrease rate of the current after the addition of two kind of OPs (terbufos and parathion), respectively. The detection limit for two kind of OPs are found to be 1 ng/mL with the linear range 1 ng/mL - 10 000 ng/mL. Cyclic voltammetry was conducted to detect parathion on the sensors which immobilized AChE by physical adsorption. The results show that it can achieve a better sensitivity than amperometric I-t curve. PMID- 17111625 TI - [Bioaccumulation of titanium dioxide nanoparticles in carp]. AB - Analysis method for TiO2 nanoparticles in water and fish samples was set up using ICP-AES after dissolved by solution of sulphuric acid and ammonium sulphate. Meanwhile bioaccumulation of TiO2 nanoparticles was assessed by tests exposing carp (Cyprinus carpio)to TiO2 nanoparticles suspensions. The relative standard deviation (RSD) for water sample of 20.0 mg/L TiO2 is 4.53%, and the recoveries of TiO2 nanoparticles in water and fish samples are raging from 94% - 104% and 90% - 103%, which could ensure accurate measurement of TiO2 nanoparticles. TiO2 nanoparticles have a significant bioaccumulation by the carp, and TiO2 nanoparticles concentrations in carp exposed to 3 mg/L and 10 mg/L TiO2 nanoparticles suspensions for 25 days are 2.1 mg/g and 5.8 mg/g respectively, and the BCF values at equilibrium are 675.5 and 595.4 respectively. Significant As and TiO2 accumulation occurs in viscus and gills of fish, while bioaccumulation of TiO2 nanoparticles in muscle is relative small. PMID- 17111626 TI - [Establishment of method for cytochrome P450 of earthworms (Eisenia fetida) as a biomarker]. AB - Earthworm (Eisenia fetida) was chosen as test soil fauna to establish the method for determination of cytochrome P450 content. By means of thoroughly washing with salt solution, changing centrifugal acceleration and dissolving microsomal membranes of earthworms with sodium cholate, the determination of cytochrome P450 was performed. Base on the method, dynamic relationships of dose-response for cytochrome P450 contents in earthworm and phenanthrene concentrations were conducted by contact filter paper and soil contaminated with phenanthrene. Results indicate that cytochrome P450 are covered up by interferential material in earthworm, which makes the classic peak lag and appear at 455 - 457nm in the CO difference spectrum with a spectrophotometer. Through increasing centrifugal acceleration and adding reagent of solubilization, the interference is removed mostly and the classic peak of P450 returns to appear at 450nm +/- 1nm. The exposure tests of phenanthrene with contact filter paper (10(-6), 10(-5), 10(-4), 10(-3) and 10(-2) mg x mL(-1)) and soil (1, 2, 4 and 8 mg x kg(-1)) reveal that the dose-response relationships exist between the pollutant and cytochrome P450. Soil tests of different exposure durations (1, 3, 7, 14 and 28d) show the effects of different phenanthrene concentration on earthworm cytochrome P450 contents are to activate (7d) and to inhibit (14d and 28d), and the content of P450 is 0.99 - 1.41 and 0.77 - 0.88 (p < 0.05) times of control respectively. From this study, cytochrome P450 content of earthworm has the merit of simpleness, speediness and economy for determination, and it could be used as a sensitive biomarker for monitoring the exposure of sublethal pollution in terrestrial ecosystem. PMID- 17111627 TI - [Joint toxicity on multi-component mixtures of SDS and substituted aromatic compounds]. AB - The toxicity to Photobacterium phosphoreum of different mixture systems of sodium dodecyl sulfate (SDS) and three substituted aromatic compounds according to equiconcentration ratio of 1 : 1, as well as individual toxicity, were determined respectively. The joint toxicity was evaluated by toxicity unit (TU), additive index (AI), similarity parameter (lambda), and mixtures toxicity index (MTI). Four evaluating methods were compared and the modes of joint action of different systems were analyzed primarily. The coherent results were obtained from four different evaluating methods. Conclusions can be drawn that the binary joint effects of phenol and toluene mixed with SDS showed weak synergistic, while the other joint effects of SDS and substituted aromatic compounds were antagonistic. Of the evaluating methods, Toxic Unit is most sensitive with higher value of its parameter. Mechanisms of joint toxicity action could be discussed according to luminescence principle of Photobacterium phosphoreum and molecule structures of the mixture components. PMID- 17111628 TI - [Cd uptake and accumulation in grains by hybrid rice in two paddy soils: interactive effect of soil type and cultivars]. AB - Cd translocation through soil-food crop-diet is considered as one of most important pathway for human Cd exposure. Rice is considered as a particular crop with high Cd uptake and accumulation in grains among the main food crops. In this study, a pot experiment was conducted to elucidate mutual interaction of soil and cultivars on uptake and grain accumulation of Cd by hybrid rice with or without Cd spiking at 2.5 mg x kg(-1) under continuous submerging condition. Two hybrid rice cultivars (Shanyou 63, a common hybrid rice and II Youming 86, a super-rice) and two paddy soils (a Wushantu, Gleyic Stagnic Anthrosols and a Hongshanitian, Ultic Stagnic Anthrosols) were used. The results show significant differences in Cd uptake and grain partitioning between soils, cultivars and the soil-cultivar interactions. The cultivars effect on uptake of indigenous soil Cd seems stronger than the soil effect while soil effect turns significant over that of cultivars on spiked Cd. However, intense Cd accumulation in grains is found under the positive interaction of soil with high Cd availability and cultivar with high Cd affinity (super rice on acidic paddy soil). This study demonstrates a phenomenon of intense Cd uptake and grain accumulation by super rice and, thus, imposing a very high Cd exposure risk (as several times as the acceptable daily intake, ADI) to subsistence-diet farmers. The low Cd cultivar Shanyou 63 tends to hamper the up-taken Cd in root while the super rice II Youming 86 promotes higher partitioning to grain. Furthermore, the difference in total biomass between the two cultivars is small compared to that in total Cd uptake under Cd spiking. It is suggested that the Cd uptake behavior should be taken into account in super rice breeding and practical measures should be taken while spread of super rice cultivars in rice areas with acidic soils and under Cd pollution in order to control the human Cd exposure by diet. PMID- 17111629 TI - [Influence of copper, cadmium on growth and cation exchange capacity of two kinds of ectomycorrhizal funguses]. AB - Ectomycorrhizal fungus has the ability to enhance the growth of higher plants in the contaminated area, especially ruined by heavy metals. And much attention was focused on how the fungus could enhance the resistance of higher plants. We focused on the resistance of ectomycorrhizal fungus in vitro to heavy metals. In the first experiment, the mycelium biomasses of two ectomycorhizal funguses growing in the Kottke media treated with different concentrations of Cu and Cd were measured after growth as well as the pH value of the medium. The results indicated that heavy metals could reduce the biomasses of the two funguses. Gomplhidius viscidus has higher tolerance to Cu but less Cd than that of Boletus edulis. With development of fungal mycila, the pH value of medium dropped significantly, and this effect might play an important role in enhancing its tolerance. In addition, the higher pH value change per biomass indicated that the fungus treated with heavy metals had the ability to adjust environment of pH more significantly. In the second experiment, the cation exchange capacity (CEC) of the cell walls of the fungus treated with heavy metals was measured according to Marschner's. The results indicated that with the increasing of the concentrations of Cu or Cd, the CEC of Gomphidius viscidus increased, but the CEC of Boletus edulis dropped. PMID- 17111630 TI - [Phytoavailable lead in rhizosphere of lettuce]. AB - A greenhouse experiment was performed to investigate the influence of pH TOC, and fractionation of Pb on phytoavailability of Pb in the rhizosphere of lettuce. And the Low molecular-weight organic acids (LMWOAs) were used to evaluate the phytoavailability of soil Pb to lettuce. The results show that the pH is lower in the rhizosphere compared to the bulk soil, and the rhizosphere becomes more acidic with increasing Pb (NO3)2 applied. In contrast, the TOC content is higher in the rhizospherer compared to the bulk soil, and more TOC accumulates in the rhizosphere with increasing Pb (NO3)2 applied. The concentrations of different fractions in rhizosphere soils follow the order of exchangeable > carbonate bound > Fe-Mn oxide bound > organic bound > residual, and for bulk soil the order is carbonate bound > Fe-Mn oxide bound > exchangeable > organic bound > residual. Compared with bulk soils, exchangeable Pb increases significantly in rhizosphere. With increasing Pb (NO3)2 applied, the transformation of residual Pb to carbonate bound and exchangeable Pb may have happened and results in the accumulation of carbonate bound and exchangeable Pb. The air-drying increases the concentration of exchangeable, carbonate bound and Fe-Mn oxide bound Pb and decreases organic bound and residual Pb. Rhizosphere and bulk soils were extracted by low molecular weight organic acids(LMWOAs) to fractionate Pb fraction of soil pools, which is considered as the phytoavailable Pb. The results show that phytoavailable Pb in rhizosphere is much higher compared to bulk soil, and phytoavailable Pb increases with increasing Pb (NO3)2 applied due to acidification processes and accumulated TOC in the rhizosphere. Air-drying process also increases the phytoavailable Pb. The root-induced changes of Pb availability in the rhizosphere might also be related to the Pb species, especially carbonate bound and exchangeable Pb. Correlation analysis indicates that application of wet fresh rhizosphere soils and LMWOAs should be recommended in the future study on the phytoavailability of soil Pb to lettuce. PMID- 17111631 TI - [Effects of several low-molecular-weight organic acids on the release kinetic of DDTs from red soil]. AB - The kinetic release behavior of DDTs from red soil with three kinds of low molecular-weight (LMW) organic acids solution and water leaching was investigated by kinetic device designed. The results show that the release percentage of DDTs from red soil by LMW organic acid can increase by 15% - 18% more than that by water, especially for the initial 250 mL leaching solution. When using water as leaching solution, the data of release kinetics of DDTs from red soil conformed to the apparently first dynamics equation (R2 > 0.99, p < 0.000 1), it implied that the surface diffuse of soil mineral is primary release mechanism of DDTs from red soil in water leaching system. The kinetic release behavior of DDTs in LMW organic acid leaching system can be described by parabola diffuse equation, double constant equation or Elovich equation. It implied that the LMW organic acids induced the complication of the release mechanisms of DDTs from red soil, this maybe related to the dissolution of soil mineral surface and structure change of inherent organic material that coating onto the soil mineral surface induced by LMW organic acid. PMID- 17111632 TI - [Temporal and spatial distribution characteristics of nitrogen losses in hilly area of Taihu Lake]. AB - Four typical land uses in hilly area of Taihu Lake were studied on temporal and spatial distribution characteristics of nitrogen losses in surface runoff under natural rainfall through experiment in situ. The medium value of event mean concentration (EMC) of ammonia nitrogen (NH4(+) -N) which dominated agricultural N in surface runoff accounted for 44.5% of total nitrogen (TN), while nitrite nitrogen (NO2(-) -N) accounted for 1.8%. Concentration of nitrogen in runoff had significantly seasonal variation which was related to meteorologic conditions such as rainfall, temperature, and agricultural activities. Temporal variabilities of site mean concentration (SMC) for TN, NH4(+) -N, NO3(-) -N and NO2(-) -N were decreased sequentially. The highest SMC value of TN, NH(+) -N and NO3(-) -N in upland runoff and N2(-) -N in bamboo grove were observed. The spatial distribution of nitrogen losses was determined by fertilizer application and vegetation coverage. Spatial variabilities for SMC of nitrogen were less than temporal variabilities. It was found that transportation fluxes of nitrogen in surface runoff from upland and vegetable plot were higher than that from chestnut orchard and bamboo forest which have significantly related to surface runoff volume. PMID- 17111633 TI - [Spatial variability analysis of soil arsenic in Zibo maize producing area, Shandong Province]. AB - Spatial variability and distribution of soil arsenic in Zibo maize producing area, Shandong Province were studied using geostatistics and geographic information system (GIS) techniques. Positive skewed distribution was observed; Logarithm transformation was applied in order to achieve normality in the data set and to reduce the affect of outliers. The results show that experimental semi variogram exists distinct anisotropic and has the strongest continuity in the direction of northeastern 70 degrees. Exponential model was used to fit experimental semi-variogram. Normal kriging was used to predict the spatial distribution of soil arsenic in Zibo maize producing area. It can be concluded that there is clear special heterogeneity in soil arsenic and wastewater irrigation is the main reason affected spatial variability. In fact, the concentrations of soil arsenic in Zibo maize producing area decrease in the pass 20 years. PMID- 17111634 TI - [Influence of temperature on hydrolysis and acidogenesis of kitchen wastes in two phase anaerobic digestion]. AB - The experiment evaluated the solubilization rate and VFA production rate of kitchen wastes with controlled-temperature at 25 degrees C, 37 degrees C, 40 degrees C and 50 degrees C, and subsequently investigated the lactic acid production and constituents of individual organic acids at various temperature levels. Results show that the increase of temperature when below 37 degrees C may improve both hydrolysis and acidogenesis rates and acidogenesis rate decreases while hydrolysis rate increases above 37 degrees C. The maximum VFA concentration of 34.4 g x L(-1) was achieved at 37 degrees C and a higher hydrolysis rate of 82% was obtained at 50 degrees C. Formic and acetic acid were predominant VFA, while propionic and butyric acid took a relatively little part. And lactic acid concentration remained relatively high. Accroding to the experiment an optimum temperature to optimize both hydrolysis and acidification is 37 degrees C. PMID- 17111635 TI - [Kinetic of pH control in anaerobic digestion of organic fraction of municipal solid waste in a batch reactor]. AB - Using a material and ionization balance analysis of anaerobic digestion process, a kinetic model of pH control in a batch anaerobic digestion of organic fraction of municipal solid waste was established on the basis of substrate decay and microbial growth kinetics, and a corresponding computer soft ware was created. The optimal pH in different anaerobic digestion can be predicted by this model. Consequently the maximal methane production can be obtained in anaerobic system by controlling the pH in optimal value. Comparative experiments were conducted to validate the model. The experiments demonstrated that the methane production of anaerobic system under optimal pH was steadier than the same condition under uncontrolled pH, and the cumulative methane production had an average increment about 20%. PMID- 17111636 TI - [Immobilization technology and mechanism of fly ash using H3PO4]. AB - Chemical composition and toxicity leaching characteristics of fly ash was analyzed. The experiment results show that many heavy metals were contained; leaching concentration of Pb is 67.03 mg/L, which exceeds the limit of identification standard for hazardous wastes. Effect of input mass of H3PO4 on immobilization of heavy metals and its long-term environmental stability was studied. The results show that when input 8% - 14% (H3PO4 mass/ fly ash mass) of H3PO4 sound immobilization effect can be achieved; 8% and 12% of H3PO4 will bring a satisfactory environmental stability of heavy metals, while more H3PO4 led to less buffer capacity to acid conditions. In fly ash treated by 12% H3PO4, a small quantity of crystal Cr2P2O7, ZnP2, Pb3P4O13, Pb3P2O7, NaZnPO4, NaPbP3O9, Ca2ZnSi2O7 can be detected by XRD; many independent fly ash particles and bar shaped Pb5 (PO4)3Cl with a diameter of 0.3 - 0.5 microm were observed by SEM; concentrated heavy metal materials were not obtained by CHBr3 floatation. Conclusions can be drawn that, through neutralization reaction of H3PO4 with strongly alkaline fly ash, stabilization reaction conditions were improved, entrapped heavy metals were chemically activated and PO4(3-) needed in stabilization was produced. Activated heavy metals combined with PO4(3-) on surface of fly ash,generated phosphates existing as forms of solid solution in SiO2, CaCO3, CaSO4, KCl, NaCl. PMID- 17111637 TI - HHS opens doors to greater interoperability of EMRs. AB - Interoperability provides real-time exchange of vital patient information that can impact quality, safety. Organizations with well-established EMRs cite significant benefits to performance improvement. Quality managers should be part of system selection process, to ensure it has the capabilities they require. PMID- 17111638 TI - ICU uses 'bundles' to make huge improvements. AB - Bundling concept based on grouping of evidence-based best practices. Each bundle includes a specified number of steps that should be followed for all patients. Daily monitoring to ensure compliance is a critical component of a successful program. PMID- 17111639 TI - Staff education, 'tough love' key to RRT success. AB - ICU decreases cardiac arrests per 1,000 discharges by 44%. It's important to establish call criteria for rapid response team early on in the program. Intense staff education and culture change are challenging but absolutely essential. PMID- 17111640 TI - Med students can aid in safety improvement. AB - Study details four cases where med students helped prevent--or could have helped prevent--medical errors. Harm prevented includes averting non-sterile conditions, missing medications, mitigating exposure to highly contagious patients, and respecting DNRs. Formal mechanism for students to voice concerns should be implemented. PMID- 17111641 TI - ACE aims to help lower performing facilities. AB - Letters may be subtle reminder of potential consequences down the road. Recipients "should make strenuous efforts to be self-critical and improve and to ask for help," expert says. Greatest challenge in performance improvement is getting essential people on board. PMID- 17111642 TI - 'Ideal patient day' gives heads-up on care. PMID- 17111643 TI - Losing the brain. PMID- 17111644 TI - I'm not crazy, I'm just depressed: differential diagnosis of dementia vs. depression. PMID- 17111645 TI - Personal experiences in home based care of the dementia client. PMID- 17111646 TI - Self-neglect: Diogenes syndrome and dementia. PMID- 17111647 TI - Montessori-based dementia care. AB - Montessori-based Dementia Care is an approach used in Alzheimer's care that does not involve chemical or physical restraints. This program works by giving the elder with Alzheimer/Dementia a purpose by getting them involved. When staff/families care for a confused Alzheimer/Dementia patient, who is having behaviors, the Montessori program teaches them to look at what is causing the behavior. When assessing the elder to determine what is causing the behavior, the goal is to find the answer, but the answer cannot be dementia. The goal of the program is to bring meaning to the life of an Alzheimer/Dementia elder. PMID- 17111648 TI - [Prevention of cardiovascular diseases: collaborative program]. AB - The so-called Program for preventive collaboration model joins the expertise of specialists and GPs and as such allocates for the effective, cost saving, secure screening of individuals with high and intermediate cardiovascular risk and identifying asymptomatic patients within a big population. Based on SCORE risk assessment GPs define their patients cardiovascular risk. In Gottsegen Gyorgy Institute of Hungarian Cardiology on patients with intermediate or high risk ankle-brachial doppler index, carotis intima media thickness, artery stiffness, microalbuminuria and left ventricular hypertrophy are diagnosed. Based on the results of non-invasive cardiovascular marker tests a mixed score is elaborated. The applicability of the score is then measured for the further risk assessment of individuals with intermediate or high CV risk. The collaboration of GPs and specialists along with the non-invasive cardiological tests are helpful in the early identification of patients with high cardiovascular risk and when making a decision about drug or non-drug treatments. PMID- 17111649 TI - [Questions of dermatoinfectology in the practice]. AB - The importance of bacterial, viral and fungal diseases has significantly increased during the past decades. The reasons are numerous, but the most important ones are as follows: appearance of new variance of microbes, appearance and spread of antibiotic resistant bacterial strains, and increasing number of patients with various degree of immunodeficiency. For such reasons we consider extremely important to overview and upgrade our current knowledge and practice regarding to these diseases. This manuscript will discuss the hottest practical questions of dermato-infectology. PMID- 17111650 TI - [Influence of diabetes mellitus on cerebral ischemia and reperfusion injury]. AB - Cerebral ischemia, caused by disturbance of the blood supply to the brain, is a major cause of death in our days. Diabetes mellitus exacerbates neuronal death induced by an ischemic insult. It is important to characterize the underlying mechanism of the cell damage in order to design therapeutic agents. The purpose of this study is to summarize some of the intracellular events leading to aggravated cell injury after diabetic ischemia including mitochondrial dysfunction. Release of mitochondrial cytochrome c activates the cell death executioner caspase-3 protease resulting in the cleavage of poly-ADP ribose polymerase (PARP) involved in DNA repair. Mitochondrial dysfunction is associated with enhanced production of free radicals such as superoxide anion, nitric oxide and peroxynitrite after diabetic ischemic injury. Mitochondrial dysfunction affects not only neurons but also astrocytes, which play an important role in neuronal functions. Damage of these cells participates in the exaggerated brain damage after cerebral ischemia. In summary, diabetes mellitus enhances intracellular pathways activated by cerebral ischemia and leads to exaggerated brain damage in diabetic subjects. PMID- 17111651 TI - [Water- and electrolyte secretion by salivary glands. ]. AB - INTRODUCTION: In salivary glands, fluid transport is thought to be driven osmotically in response to transepithelial salt gradients. According to the classical two-stage hypothesis of salivary secretion, an isotonic primary fluid is generated by the acinar cells and the fluid is subsequently modified by solute reabsorption and secretion as it passes along the ductal system resulted in the final, hypotonic solution. AIM: Very little is known about the molecular and functional nature of the transporters involved in salivary secretion, especially in human salivary glands. Therefore a systematic investigation of membrane transporters expressed also in the kidney, has been undertaken in healthy human salivary glands. METHODS: RT-PCR and immunohistochemical analysis of different membrane transport proteins was used in rat and human salivary glands. RESULTS: Clear evidence for the expression of aquaporin water channels in human salivary glands was found. AQP1 in the myoepithelial cells, AQP3 in the basolateral, AQP5 in the apical membrane of the acini is localized. The electroneutral NBC3 Na(+) HCO3(-)-cotransporter is present in the apical membrane of the serous acini and of the ducts, while the NBCn1 only in the basolateral membrane of the striated duct is localized. The NHE1 Na+/H+ exchanger is present in the basolateral membrane of the acini and ducts. The vacuolar H(+)-ATPase is localized apically in the ducts, except for the intercalated duct. CONCLUSION: Aquaporin water channels are likely to be involved in water secretion. The NBC3 and NBCn1 electroneutral Na(+)-HCO3(-)-cotransporters, the NHE1 Na+/H+ exchanger and the vacuolar H(+)-ATPase may play an important role in the pH regulation of salivary acinar and duct cells. PMID- 17111652 TI - [Intrauterine intestinal volvulus and fetus compressus papyraceus in twins]. AB - Intrauterine intestinal volvulus and fetus compressus papyraceus in twins. In a double twin pregnancy revealed on the 16th gestation week by ultrasound examination, a foetus papyraceus was detected on the 20th gestation week in a 29 year old woman. On the 32nd gestational week two monochorionic-diamniotic male stillborn twins were born. One of them (15 cm/35 g) was rudimentary developed and compressed. On examination of the other (39 cm/1485 g) partially macerated foetus, volvulus of small intestine with hemorrhagic necrosis was found at the autopsy. Both of the anomalies are very rare. PMID- 17111653 TI - [Functional morphology of the surface epithelium of esophageal tunica mucosa]. AB - This review presents the systematized summary of classical and current conceptions on the functional morphology of the surface epithelium of esophageal tunica mucosa. The data describing the architecture of epithelial lining, classification and structure of its layers, are presented. The detailed characteristics of the cells of each layer and their ultrastructure, organization of germinal compartment, stem cell distribution and activity as related to their topography, are presented. The parameters of epitheliocyte cell cycle, mechanisms controlling their proliferation and circadian rhythms by growth factors and hormones, changes of proliferation activity under natural, pathological and experimental conditions, are discussed. The process of epithelial desquamation is described, as well as a mucus layer covering the epithelial surface together with its sources and protective role. The characteristics of the process of epitheliocyte apoptosis and the mechanisms of its control in normal and pathological states, are presented. PMID- 17111654 TI - [Interconnection of morpho-functional changes at various levels of the cortical bone hierarchic organization in aging]. AB - The authors define two groups of levels of cortical bone hierarchic organization. The first three levels (molecular, supra-molecular and tissue) are characterized by age-dependent changes in connections and geometry of collagen fibers, deviations in orientation and interaction between collagen and minerals, increase in crystallinity and size of the latter, resulting in matrix hypermineralization. It causes water and organic fraction displacement, as well as intermolecular space reduction, which provides for lesser amount of matrix deformations under the influence of mechanical forces. At the next three levels (structural functional, organ-forming structures, organ) adaptation processes contribute to deformation increase due to a greater volume of cavities (Haversian canals and medullar cavity of the diaphysis of long tubular bones). These are due to the fact that osseous tissue cells possess superior and inferior thresholds of deformation perception, and through modeling/remodeling provide for extracellular matrix migration in the direction limited by these thresholds. Bone geometry changes leading to bone mass loss are also caused by age-dependent rise of the inferior threshold of sensitivity to mechanical impulses and decrease of muscle functional activity. Prevention of the described changes may be possible through elaboration of new, pathogenesis-based ways of drug therapy, including 1) osseous tissue mineralization reduction with the help of osteocyte pump regulators for predominant washing out of Ca2+, 2) lowering the threshold of electric impulses initiation arising in the osteocyte network under mechanical deformations. PMID- 17111655 TI - [Effect of gender and neonatal androgenization on dendroarchitectonics of neurons in the dorsomedial nucleus of amygdala]. AB - The aim of this study was to establish gender-associated differences in dendroarchitectonics of neurons in dorsomedial nucleus of amygdala and the role of androgens in their formation during the period of sexual differentiation of the brain. Using Golgi method, it was demonstrated that the quantitative characteristics of long-axon sparsely branched neurons of all classes- neuroblastoform, short-dendrite and reticular--reflected the influence of gender. Specifically, it was detected that long-axon sparsely branched neurons had more branching primary dendrites and greater total dendrite length in adult males as compared to females. In adult females, androgenized neonatally by injection of 1250 mg of testosterone propionate on their postnatal day 5, the neuronal characteristics were different from those in normal females, and these differences were even more pronounced in comparison with males. PMID- 17111656 TI - [Nitric oxide synthase in human intracardiac ganglia in the normal and ischemic myocardium]. AB - Nitric oxide synthase (NOS) expression in human intracardiac ganglia was studied using two techniques--histochemical demonstration of NADPH-diaphorase and immunohistochemical staining for NOS. To detect the influence of coronary heart disease on NOS expression, hearts were studied in patients that died from heart failure (n = 8) and in persons that died in accidents (n = 3, control). It was found that human intracardiac neurons normally expressed mainly NOS1, and the proportion of these cells amounted to about 40%. A portion of neurons with low and moderate density of staining for NADPH-diaphorase was increased in ischemic myocardium, probably, due NOS2 induction. PMID- 17111657 TI - [Ultrastructure of optic nerve fibers in rabbits after intravitreal infusion of perfluoroorganic compounds]. AB - Ultrastructure of optic nerve myelinated fibers after intravitreal infusion of perfluorooctylbromide (PFOB) and perfluorodecaline (PFD) for 30 days was studied in rabbits (n = 14). After inraocular administration of PFOB, the ultrastructural changes of optic nerve fibers were mainly reactive in nature. Mitochondrial swelling and partial destruction of cristae were detected in the axons. After PFD infusion, the changes were observed that were not only reactive, but highly degenerative, including destruction of organelles and myelin sheath and axon cytoskeleton disintegration. Thus, the changes associated with PFOB were less pronounced than those found after PFD application. Therefore, PFOB is more promising for the use as an ophthalmologic implant. PMID- 17111658 TI - [Electron-tomographic analysis of the Golgi complex structure in cultured cells]. AB - Study of the Golgi complex (GC) in cultured NRK cells using improved method of sample preparation for electron tomography (ET) enabled to detect more fine details of GC structure. With the application of quantitative ET, no numerous vesicles were detected around GC, while most of stacks constantly contained intercisternal connections. It was demonstrated that vesicular-tubular clusters, which serve as the exit sites from endoplasmic reticulum, were composed of two domains: varicose tubules with sparse COPII-coated vesicles and a network of smooth tubules. The data obtained argue against the models of transport, which consider the vesicles as the main or necessary membranous carriers. On the contrary, continuous tubular connections seem to play an important role in traffic joining secretory pathway regions. PMID- 17111659 TI - [Effect of radiation sterilization on biomaterial structure and properties]. AB - The purpose of the investigation was the study of structural changes connective tissue biomaterials after different combinations of their radiation sterilization, physico-chemical treatment and conservation. Using a complex of histological methods (polarization microscopy of unstained sections, van Gieson's stain, scanning electron microscopy), an analysis was performed to detect the structural changes in tendons and dermis which underwent various physico-chemical treatment and subjected to the radiation sterilization using different types (3- radiation and fast-moving electron stream) and doses (1.5, 2.5 and 4 Mrad) of radiation. The dependence of the structural changes in the biomaterials on the fibroarchitectonics of the tissue as well as on the physico-chemical treatment and radiation type and dose was demonstrated. The tendons underwent clear-cut significant changes in all regimes and radiation doses studied. Dermis was found to be most resistant to radiation challenge. PMID- 17111660 TI - [Changes in myocardium, skeletal muscle and liver of rats fed carnitine-deficient diet and treated with carnitine optical isomers]. AB - The aim of the present study was a comparative assessment of L-, D-and DL carnitine effect on morphometric and histological parameters of myocardium, skeletal muscles (m. gastrocnemius) and liver in 60 rats fed carnitine-deficient diet. Carnitine-deficient diet fed 2 months resulted in a substantial reduction of carnitine concentration in blood plasma of rats. In carnitine-deficient animals, lipid vacuoles were found to accumulate within the hepatocytes in all the zones of hepatic lobules, which mainly had the character of micro- and macrovesicular steatosis. This was accompanied by a reduction of skeletal muscle fiber and cardiomyocyte average thickness. L-carnitine administration resulted in the compensation of carnitine deficiency in animals with alimentary carnitine deficient state, while the racemate and D-stereoisomere did not affect its content in blood. Pharmacological correction of carnitine deficiency with L carnitine prevented the development of liver fatty dystrophy to a greater degree, than the administration of other carnitine stereoisomeres and promoted the restoration of muscular fiber thickness of skeletal muscles. DL-carnitine administration was accompanied by a moderate correction of fatty dystrophy and did not prevent the development of skeletal muscles atrophy. D-carnitine stereoisomere did not prevent liver fatty dystrophy, but it reduced its severity. Correction of carnitine deficiency with D- stereoisomere was not accompanied by essential morphological and morphometric differences in degree of skeletal muscle atrophy. PMID- 17111661 TI - [Ultrastructural characteristics of endotheliocytes of skeletal muscle hemomicrocirculatory bed during tibial lengthening using G. A. Ilizarov technique]. AB - The method of transmission electron microscopy was used to study the ultrastructure of endotheliocytes in the hemomicrocirculatory bed of the anterior tibial muscle of adult dogs during tibial lengthening with Ilizarov method. The capacity of endotheliocytes to reduction and renewal was established. The tension stress induced in the biological tissues of the lengthened limb stimulates and supports high activity of endotheliocyte nuclear apparatus, which is characterized by the prevalence of euchromatin. Presence of capillaries with closed lumen at all time points studied is indicative of a prolonged angiogenesis process in the terminal parts of the hemomicrocirculatory bed, which is accompanied by regeneration and growth of the muscle fibers. After the cessation of distraction, while the muscle tissue continued to grow and differentiate, the endotheliocyte growth and the activation of their synthetic apparatus was noted. PMID- 17111662 TI - [Morpho-functional characteristic of the spleen in Baikal seal (Pusica sibirica Gmel.) pups]. AB - Using the methods of light microscopy, the histological structure of the spleen in Baikal seal (Pusica sibirica Gmel.) pups was studied for the first time. It was found that in Baikal seal the spleen is characterized by highly developed supporting elements (thick capsule and numerous branching trabeculae), which are rich in smooth myocytes. By the age of 2-4 weeks, all the structures of white pulp were formed, however, presence of only a few lymphoid nodules with the absent germinal centers indicates that functionally these structures are still immature. Presence of immature forms of granulocytic, erythroid and megakaryocytic lineage shows that the myelopoiesis in Baikal seal spleen continues in postnatal period. PMID- 17111663 TI - [The histogenesis of interrenal primordium of the adrenal gland in pig (Sus domestica)]. AB - Using light, electron microscopy and cytochemistry, the early (embryonic week 4 8) stages of adrenal gland (AG) development were studied in domestic pig. The interrelations between the cells of the fetal cortex (FC) and chromaffin cells (CC) were traced. At week 5, AG primordium is represented by FC, which consists of the epithelioid cells, with the ingrowing neural cords containing CC islets. Starting at the early embryonic period and up to fetal period, CC and interrenal cells of FC are closely interrelated with each other and sinusoidal capillaries. Both cellular types are at different stages of differentiation, including the functionally active elements. At weeks 7-8, FC cells undergo involution, while those ones, left at periphery, form definitive cortex. CC are located in the central part of the organ and form suprarenal tissue. Authors hypothesize, that CC, migrating into AG primordium, initially induce the development of interrenal primordium, and later cause the involution of FC. This, possibly, may be explained by the fact that further antenatal and postnatal development of the organism requires more corticosteroids than the amount produced by FC. PMID- 17111664 TI - [Organization and cytochemical features of barrier structures in human placenta]. AB - The aim of the present study was to investigate structural pattern of human placental barrier elements using light and electron microscopy and immunocytochemistry. Some important peculiarities of organization of the placental barrier were detected: difference in structure and amount of collagen IV in the basal lamina of endothelium and trophoblast, occurrence of smooth muscle actin in the capillary wall forming syncytiocapillary membranes. In the intercapillary stroma of terminal villi, both fibroblasts and macrophages but not myofibroblasts were found. Since smooth muscle cells and myofibroblasts are absent, pericytes are most likely cells to contain smooth muscle actin in the area of syncytiocapillary membranes. PMID- 17111665 TI - [Body component composition of healthy children and the patients of psychoneurological child houses for children of the early age]. AB - The aim of the study was to evaluate body component composition of healthy and ill children starting from their birth up to 3 years. The results of skin fold caliper measurements of 272 healthy children and 357 children with perinatal encephalopathy are presented. In healthy children, the ratio of lean and fat mass is equal to 4.0. Fat depots of girls are larger than those of boys. In ill children, body fat reserves are diminished and sex-related differences of body composition are absent. The shorter was the period of intrauterine development, the lower is the fat content in the child's body. Among the ill children, the best indexes of body composition were found in patients with hypoxic encephalopathy, and the worst ones--in children with fetal alcohol syndrome. PMID- 17111666 TI - [Periodical pattern of cytochrome oxidase activity distribution in the visual cortex of kittens]. AB - The aim of this study was to determine the distribution of cytochrome oxidase activity in the visual cortex of normal kittens. Method of computer filtration of digital images of frontal sections was used. The periodicity of enzyme activity in layers III, IV, VI of area 17 and in layers IV, VI of area 18 has been shown. PMID- 17111667 TI - [How to measure structures, or new stereology: III. Stereology and electron microscopy]. AB - This review describes modern stereological approaches and methods for estimation of subcellular structures dimensions. These estimators could be used for determination of number- and volume-weighted parameters of particle size. Very often they do not require the application of the disector at the first stage of sampling of images. Additionally, the modified version of the rotator for point counting is demonstrated, which significantly accelerates the morphometric procedure. PMID- 17111668 TI - [Potentialities of intravital methods of the evaluation of morphometric pelvic characteristics in adult persons]. AB - Comparative assessment of posthumous and intravital methods of morphometric study of the pelvis in adult persons was performed in 110 embalmed anatomical specimens and in 81 patient. It was proved statistically, that magnetic resonance tomography and spiral computer tomography (SCT), performed following definite algorithm, appear to be highly informative methods for the evaluation of pelvimetric parameters. They permit to define various osseous landmarks with high precision and to perform the measurements of linear metric pelvic parameters. On the basis of the data obtained, various pelvimetric indices that characterize bony pelvis shape, could be reliably calculated. Moreover, SCT together with shade surface reconstruction in different projections permits to demonstrate visually the 3D shape of the pelvis or its separate fragments and to evaluate the symmetry, presence or absence of bone structural deformations. PMID- 17111669 TI - [New conceptions of structural organization of the active lymph drainage]. AB - The review examines the history and the current state of the problem of structural bases of active lymph drainage from organs in human and mammalian animals. Two conceptions of lymphangion structure are compared that represent it as either valvular or intervalvular segments of lymphatic vessel functioning as active lymph drainage organizers. PMID- 17111670 TI - [Morphological and biochemical aspects of skeletal muscle injury and regeneration subjected to physical load and hypodynamia]. AB - This paper reviews the literature data on morphological and biochemical aspects of skeletal muscle injury by exercises, hypodynamia and microgravity. Muscle injury depends on the duration and intensity of action. In spite of differences of muscle injury mechanisms by exercises and hypodynamia, this injury restricts muscle function and capacity to continue muscle work. Possible approaches to minimization of the muscular tissue injury and accelerating its regeneration are discussed. PMID- 17111671 TI - [Role of competitions in normal anatomy in enhanced motivation to study the subject]. PMID- 17111676 TI - A more effective prescription for ATV safety. PMID- 17111677 TI - Traumatic vertebral artery dissection and stroke in a 16-year-old male as the result of an ATV accident. PMID- 17111678 TI - Mortality rates among chemical workers in the Kanawha Valley of West Virginia: 1940-1999. AB - To expand a cohort of chemical workers in the Kanawha Valley, we conducted a study of 33,225 workers who were employed at three locations between 1940 and 1999. We observed no increase in overall cancer mortality. Higher risk estimates were observed for lymphosarcoma and reticulosarcoma among hourly employees who worked at the Institute or South Charleston locations. This finding was limited to men hired before 1960. We observed no new cases of angiosarcoma of the liver, a cause of death previously reported in association with vinyl chloride production at the South Charleston location. Specific risk factors for lymphosarcoma and reticulosarcoma have not been identified in previous targeted studies of this population and it is unlikely that additional cause-specific research will elucidate the etiology. Updates of the entire cohort will continue and can be used as a comparison population for chemical specific studies within these three plants. PMID- 17111679 TI - Malignant optic nerve glioma (glioblastoma multiforme): A case report and literature review. AB - Primary gliomas of the optic nerve are very rare. We report a case of a 59-year old male with sudden vision loss diagnosed with malignant optic nerve glioblastoma multiforme. Magnetic resonance imaging revealed thickening of optic tracts, chiasm, and hypothalamus. Histologically, the tumor was composed of glial cells with pleomorphic nuclei and areas of vascular proliferation and necrosis. The patient died eight weeks after initial presentation. In addition to our case, 30 previously reported cases of malignant optic nerve glioma are reviewed. PMID- 17111680 TI - Ethylene glycol intoxication: Disparate findings of immediate versus delayed presentation. AB - Ethylene glycol is a common household substance responsible for a large number of ingestions in the U.S. each year. In 2001, nearly 5,000 ethylene glycol exposures were reported with more than 1,600 patients requiring medical treatment. There were 16 deaths attributed to ethylene glycol in 2001, second only to ethanol overdose for lethal ingestions. Diagnosis of ethylene glycol ingestion is relatively straight-forward when an individual with a history of exposure is found to have a high anion-gap metabolic acidosis and an elevated osmolar gap. Appropriate treatment can be immediately employed and the diagnosis confirmed by the finding of elevated ethylene glycol levels in the serum. In the absence of exposure history, the differential diagnosis of a high anion-gap metabolic acidosis and an elevated osmolar gap will also lead to consideration of ethylene glycol ingestion. This well-recognized presentation of ethylene glycol toxicity includes findings expected in individuals who present for care soon after their ingestion. A less well-known pattern may be seen in those for whom care is delayed. We present a patient with delayed presentation of ethylene glycol ingestion and review the physiology and biochemistry that underlies this different presentation. Unfortunately, without history or strong laboratory evidence, ethylene glycol ingestion may be easily overlooked in individuals with delayed presentation. PMID- 17111681 TI - Strategic National Stockpile--what is it? PMID- 17111682 TI - [Carbohydrate-deficient transferrin as the marker of alcohol abuse]. AB - The quantity of usual alcohol consumption is a fact that can be determined from the anamnesis of patients. In many cases patients do not inform their doctors in detail, in some cases they want to keep it a secret. The diagnosis of alcohol abuse based on objective data is a necessary requirement in clinical, insurance, as well as in forensic medicine. Among the different biomarkers of chronic alcohol abuse, carbohydrate-deficient transferrin is recognized world wide as the most reliable indicator. The authors review the pathomechanism of carbohydrate deficient transferrin in the human organism, the methods for its measurement and its role in the diagnostic procedures of alcohol induced clinical diseases, as well as its decisive role for life insurance and in forensic medicine. PMID- 17111683 TI - [Catch fingertip support in microsurgery to reduce the tremor]. AB - INTRODUCTION: With microsurgical operations one of the main risk factors is physiological tremor. To reduce it, microsurgeons usually fix their forearms and hands up to the fingertips IV-V. on supporting desks (armrests), and on the bone (skull) and skin being operated on. AIM: To improve further microsurgical technique which reduce the tremor. METHODS: The new technique gives microsurgeons support for fingertips I. II. and III. due to the "Bethlehem bridge" that can be placed quite close to the site of operation. RESULTS: An approximately tenfold reduction of tremor can be achieved due to the fixation of the crucial I. II. and III. fingertips, which hold the operating instruments. CONCLUSION: The microsurgical operations could be performed at higher precision level. PMID- 17111684 TI - [Urticaria and Quincke's edema]. AB - Urticaria and Quincke's edema is the most common cutaneous disease encountered in general medicine and dermatology. The symptoms are caused by vasoactive mediators released by mast cells following immunological or nonimmunological stimuli. Diagnosing the disease can be difficult, especially if the symptoms are chronically present and only barely react to treatment. Authors review the etiology, pathogenesis, special forms and treatment of acute and chronic urticaria and Quincke's edema. PMID- 17111685 TI - [The effectiveness of carvedilol in heart failure]. AB - BACKGROUND: The third generation beta-blocker (carvedilol) is effective in reduction of hypertension, and of mortality and morbidity as a supplement to conventional drugs of heart failure therapies (diuretics, ACE inhibitors), based on randomized controlled trials and retrospective analysis. OBJECTIVE: To analyse the efficacy of carvedilol in the treatment of heart failure with special focused on morbidity, mortality endpoints. METHODS: We assessed the multicenter, randomised, double-blind studies involving more than 150 patients (1995-2005) from MEDLINE database, in which carvedilol was used in the case of moderate to severe heart failure. We also present the results of health-economic publications (2000-2005). RESULTS: In U.S. Carvedilol Heart Failure Study (n 1096) the mortality declined by 65% (3.2% vs. 7.8%; p <0.001) with carvedilol vs. placebo, while the cardiovascular hospitalization decline was 27% (14.1% vs. 19.6%; p = 0.036) in heart failure (LVEF < or = 5%) applied together with the basic therapy (diuretic and ACE-inhibitor). In the COPERNICUS trial the efficacy of carvedilol was compared to placebo in the case of severe HF patients (LVEF < 25%, n = 2889). The annual mortality risk declined by 35% (19.7% vs. 12.8%, 95% CI 19-48%, p = 0.00013) while the risk of mortality or any risk of hospitalisation by 24% (p = 0.00004) in the active group. The CAPRICORN study (LVEF < or = 0%, n=1959) showed that carvedilol is efficacious in reduction of total (HR: 0.77; 95% CI 0.60-0.98; p = 0.031) and cardiovascular mortality (HR: 0.75; 95% CI 0.58-0.96; p = 0.024) as far as high-risk patients are concerned. CONCLUSION: The effectiveness of carvedilol is certified in reduction of mortality and hospitalization in the treatment of moderate-severe heart-failure as part of the combination therapy. The benefits of use of the drug are well measurable not only on the level of patients but on the suppliers and the financer as well, thanks to the decline of resource utilization. PMID- 17111686 TI - [Double-balloon endoscopy for the diagnosis and treatment of small intestinal disease: an initial experience from 25 examinations]. AB - Until recently, only the proximal small bowel was accessible for diagnostic and therapeutic endoscopy. Endoscopic evaluation of this organ has often required open laparotomy with surgically assisted passage of the endoscope through the intestine. Recently, Yamamoto et al have developed a new method, double-balloon endoscopy (DBE) that allows high-resolution visualization and therapeutic interventions in all segments of the GI tract. Our aim was to report our early experience with the Fujinon EN-450 T5 therapeutic double-balloon endoscope. PATIENTS AND METHODS: Between August 2005 and March 2006, 25 DBE was conducted in 23 consecutive patients (M/F: 13/10, age: 51.8 +/- 16.5 years) presenting at our tertiary referral hospitals (17 and 4 patients from the oral or the anal route, respectively; 2 patients from both). All procedures were done by i.v. anesthesia, at our outpatient clinic. After the procedure, the patients were monitored in a recovery room for at least 4h before discharge. RESULTS: The main indication for DBE was suspected small-bowel GI bleeding (11), diagnosis or complications of IBD (7), polyposis syndrome (3), stenosis (1) and insertion of jejunal catheter in one case. Twelve out of 22 patients (54.5%) had a small-bowel finding, with 16 of 22 (72.7%) of the patients having a more accurate diagnostic input. The average insertion length was app. 165 cm (range 50-350 cm, SD 97). Patients' tolerance of the procedure was excellent. No severe complications were recognized. CONCLUSIONS: Based on our limited experience double-balloon enteroscopy is a safe and useful method to evaluate and treating small bowel disease in selected patients, including patients with suspected small-bowel strictures, in whom capsule endoscopy is contraindicated. PMID- 17111687 TI - [Alternative therapy in childhood cancer]. AB - INTRODUCTION: In Hungary there are as many as 300 children diagnosed with malignant tumor each year. Along with the traditional treatments, alternative remedies have been increasingly applied in the past two decades despite the availability of the more complex and more effective malignant tumor treatments. AIMS: The authors attempted to find out about the applied alternative remedies for childhood cancer outside the traditional treatments, to determine how frequently they were applied and to examine what psychological factors had lead to their application. PATIENTS: Children's parents, getting active treatment and also the ones that have been rehabilitated at the II. Department of Pediatrics, Semmelweis University. RESULTS: 60 percent of the 34 children being examined were getting several kinds of therapy. The most frequently applied alternative remedies were the following: nature products, diet supplements, herbs, diet changes. The parents' choice was mainly influenced by other parents' opinions (42%), frequently appearing commercials in media (26%) and their current financial background (23%). The authors have found on the base of their research that the general aim of parents was: to help their own incapability to help (31%), to make sure about the efficiency of the treatment (45%). CONSEQUENCES: Independently of demographic or clinical facts the use of alternative remedies are excessively widespread and popular with children having tumor. Thus it is not advisable or reasonable to absolutely refuse them since they are quite widespread and people tend to strongly believe in them. As for doctors, they are to give professional advice and help with choosing the alternative remedies that are not harmful or even more, the ones that can make traditional treatments more effective. PMID- 17111688 TI - [Verzar Gyula (1886-1960)]. PMID- 17111689 TI - So many forces converge to undermine wellness efforts. PMID- 17111690 TI - Bush uses bully pulpit to promote transparency. PMID- 17111691 TI - Medco, consumers union face the music and dance. PMID- 17111692 TI - Physician salaries outpace production in 2006. PMID- 17111693 TI - Satisfying the P and T committee's need for more evidence. PMID- 17111695 TI - Tough bioethical questions are a slow train coming. Interview by Patrick Mullen. PMID- 17111694 TI - The logjam went data way. PMID- 17111696 TI - So you want to be a manager? PMID- 17111697 TI - Tightly organized doctors seen as best bet for quality. PMID- 17111698 TI - Evidence-based medicine gets push from businesses. PMID- 17111699 TI - Intraocular lens implants can't help but catch the eye. PMID- 17111700 TI - Health plans, employers seek common goals. PMID- 17111701 TI - [Nephrotic syndrome as a first manifestation of familial Mediterranean fever]. AB - AA amyloidosis may be a complication of Familial Mediterranean Fever (FMF). This is a case history of a female patient who did not have the classic symptoms of FMF, which usually precede the renal manifestation. The patient was admitted with edema of both legs, and the nephrotic syndrome was discovered, leading to the diagnosis of AA amyloidosis on kidney biopsy. Genetic testing uncovered the homozygous M694V type mutation, the most common mutation of FMF, which renders the patients prone to amyloidosis. This case represents the phenotype II of FMF, which presents with amyloidosis without prior classic attacks of FMF. Since effective prevention of the development of amyloidosis is available, genetic testing should be considered in order to identify mutations which carry high risk for the development of amyloidosis. This is also relevant in asymptomatic individuals with family history of FMF. PMID- 17111702 TI - [Trends and characteristics of diabetes-related lower limb amputations in the Negev, 1996-1999]. AB - BACKGROUND: The loss of a lower limb because of diabetic foot problems such as infections is an important complication of diabetes mellitus. The goals of this study were: (1) to examine trends in incidence of diabetic-related lower limb amputations in the Negev, (2) to describe the clinical characteristics of patients who underwent amputations in the Soroka University Medical Center and (3) to estimate in-hospital mortality and its predictors. METHODS: This study included all diabetic patients who underwent non-traumatic lower limb amputation in the Soroka Hospital during the period 1996-1999. The computerized hospitalization files and surgery logs during the study period were reviewed for ICD-9 diagnoses of diabetes and amputations. For each patient, hospitalization records were abstracted and data on socio-demographic and clinical characteristics were collected. RESULTS: During the study period 411 amputations were performed on 250 diabetic patients (1.6 amputation/person). The estimated mean annual incidence rate of lower limb amputations in the Negev was 5 per 1000 diabetic patients, 27.3 per 100,000 total population, and 45 per 100,000 adults above 18 years of age. The mean age was 68 (SD +/- 11.4) years. The most frequent types of surgery were standard below-knee amputation. Fourteen percent of patients died during hospitalization. Systolic blood pressure, white blood count and serum creatinine at admission were independent predictors of in-hospital mortality. CONCLUSIONS: The incidence of lower limb amputation in the Negev is similar to that reported in other countries. Interventions directed to early detection of diabetic foot problems may have an impact on the reduction of lower limb amputations and related mortality. PMID- 17111703 TI - [The value of FDG-PET/CT in the evaluation of patients with a pulmonary nodule and a previous malignant disease]. AB - BACKGROUND: Patients with primary malignant disease undergoing oncological surveillance, upon completion of their treatment, are a unique population that is gradually growing. Therefore, the number of patients with a pulmonary nodule and a previous malignant disease has increased over recent years. In order to select appropriate management for these patients, a few questions should be answered: Is it a malignant lesion? Secondly, is it primary or metastatic? Is it the only one? In the past, several imaging and invasive diagnostic procedures were used to clarify theses uncertainties. However, in many cases the results were equivocal and it was difficult to establish treatment policy. In recent years PET/CT-FDG was found to be specific, sensitive and accurate in the evaluation and staging of patients with primary malignant disease. In this study we analyzed the accuracy and value of PET/CT-FDG in patients with primary malignant disease and an indeterminate pulmonary nodule. METHODS: Patients with primary malignant disease in the past and undefined pulmonary nodule/s were recruited. Computerized retrospective screening was performed. The specificity, sensitivity, positive and negative predictive value of PET/CT-FDG were calculated. The influence of the PET results on the clinical policy was evaluated. RESULTS: Forty one patients with previous malignant disease entered the study. PET/CT-FDG was positive in 15 cases. In 13 patients (87%) the FDG positive nodule was found to be malignant. In 26 patients the PET/CT was negative. In 19 (73%) diagnosis of non-malignant nodule was confirmed. The positive and negative predictive value of PET/CT-FDG for metastatic disease was 87% and 91% respectively and the accuracy 86%. CONCLUSIONS: The results of the present study demonstrate that PET/CT-FDG is a very valuable tool in the evaluation of patients with previous primary malignant disease and a pulmonary nodule. This imaging technique was able to clarify most of the thoracic surgeon's uncertainties and provided enough data to choose the optimal treatment strategy. PMID- 17111704 TI - [Expressed emotions (EE) towards schizophrenic patients among staff members of psychiatric wards and hostels: comparison and clinical implications]. AB - Expressed emotion (EE) is an interactive criterion that describes the amount and quality of emotional communication between schizophrenic patients and their families or institutional care takers. EE is empirically related to the prognosis of the disease across cultures and therapeutic settings. This study compares results of EE assessments among psychiatric wards staff and hostels staff members. Our findings indicate that in each therapeutic milieu there is correlation between the institutional culture and the expected roles and behaviors of the patient. In the psychiatric hospital the most rejected patient is the psychotic patient presenting disturbances in thought and behavior, while the patient that is not rejected is the withdrawn non-active and not disturbing individual. In the hostel, the situation differs: the expectations from the patients are high functional levels in all fields, and therefore, the most rejected patients are those who are less functional in their initiative and occupational skills. Our findings also relate to the ability of staff members to change their attitudes towards patients in the two milieus. The findings of the comparison we performed indicate the advantages of the hostel staff's characteristics in the treatment of schizophrenia patients, and suggest specific recommendations for staff training in both milieus. The study highlights issues to be considered to ensure optimal functioning of the hostel as an appropriate setting for the treatment of schizophrenic patients. PMID- 17111705 TI - [Practice of toxicology screening in an emergency department]. AB - The present study examines standards of toxicology screening in the Emergency Department. METHODS: A retrospective analysis was performed on 583 patients for whom 713 different toxicology assays were conducted. RESULTS: The most commonly used test was the urine toxicology assay which was found positive in 10% of the subjects. Most of the patients in this group were treated with respective antidotes before the results of the tests became available. One out of 13 subjects who denied acetaminophen ingestion showed toxic blood levels. High carboxyhemoglobin saturation was found only in subjects with symptoms of intoxication. CONCLUSIONS: The medical utility of urine toxicology assay is not clear, as patients reviewed were treated according to their clinical conditions. Acetaminophen blood levels should be examined more frequently, as symptoms are vague and appropriate treatment is very effective. Carboxyhemoglobin saturation should be examined whenever exposure is suspected and symptoms and or signs are compatible with intoxication. PMID- 17111706 TI - [Non-fearful panic disorder]. AB - Panic disorder is among the most prevalent mental health disorders. Among other reasons, it is an important disorder due to its influence on all other medical professionals that these patients consult repeatedly. One of the major reasons for sub-optimal diagnosis is the different and less familiar presentations of the disorder. This article will focus on the "non-fearful" variant. Four case reports are described with prevalent presentations of the disorder and a summary of recommendations for better diagnosis of the disorder. PMID- 17111707 TI - [Treatment of pediatric femur fractures by immediate reduction and spica cast application--clinical and economical feasibility in the Israeli medical system]. AB - BACKGROUND: Pediatric femur fractures are common injuries necessitating immediate application of spica cast or surgical stabilization. Currently, neither conservative nor operative fixation shows superior results; nevertheless, operative intervention has become more common lately. The current study retrospectively investigates short term results of immediate spica cast application, as well as the economic consequences of this treatment. METHODS: All cases of pediatric (6 months - 6 years) femur fractures treated in a single medium-sized hospital during 3 years were reviewed. Characteristics of fracture treatment and economical aspects were analyzed. RESULTS: Twenty four patients (17 boys and 7 girls, mean age 2.5 years, range 9 months to 5.5 years), were treated during the study period by immediate closed reduction and spica cast application. Eleven patients were also treated for medical reasons. All cases achieved acceptable alignment and shortening after cast removal. Two patients were re admitted to the hospital Emergency Room but no changes in treatment were needed, one patient needed re-reduction. Mean hospital stay was 2.75 days. Calculated cost of such treatment is estimated to be almost equal to the compensation given by insurance companies (based on hospital stay in Israel). CONCLUSIONS: Immediate application of spica cast for pediatric femur fractures, achieves its goals of appropriate fracture-alignment and acceptable complication rates. The authors believe that the compensation should be procedure-based and not hospital-stay based, in a way that will encourage higher efficacy of medical treatment and shortened hospital stay. PMID- 17111708 TI - [Anemonia sulcata sting]. AB - A two-year-old child was stung by a sea anemone of the Sulcata type, in the scrotum. The natural habitat of this sea anemone is shallow waters in the rocks on beaches. The sting of the sea anemone is uncommon and usually causes slight local manifestations, however, in more sensitive areas of the body a stronger reaction occurs. A sting in the scrotum causes a burn with harsh local manifestations including swelling, redness and pain requiring hospitalization. The only treatment is symptomatic. In all cases recuperation is complete without any scaring. This case is unique because no outer clothing, such as the protection of a bathing suit, was used. PMID- 17111709 TI - [Herbal medicine in womens' life cycle]. AB - Women use herbs and other traditional and complementary modalities to treat various ailments throughout their life circle. This article reviewed 19 randomized controlled trials, which studied efficacy and safety of various herbs in the treatment of premenstrual syndrome (PMS), nausea and vomiting in the first trimester of pregnancy and menopausal hot flushes. Preliminary data support the efficacy of Chaste tree fruit (Vitex agnus) in the treatment of PMS, Ginger (Zingiber officinale) in the treatment of hyperemesis gravidarum and (Cimicifuga racemosa) in the treatment of menopausal hot flushes. Additional and more rigorous studies are warranted in order to support the efficacy and safety of these herbal remedies. PMID- 17111710 TI - [Amyloidosis of familial Mediterranean fever (FMF)--insights to FMF phenotype II]. AB - Amyloidosis is the most grievous manifestation of Familial Mediterranean Fever (FMF), occurring in a high proportion of untreated patients. Continuously elevated serum amyloid A (SAA) levels during remissions, rather than a pulsatile rise during FMF attacks, underlies the development of amyloidosis. FMF phenotype II is one extreme of AA amyloidosis, evolving despite a complete absence of FMF attacks. FMF phenotype II is diagnosed in patients with AA amyloidosis in the context of a family history of FMF. In these patients and in patients with AA amyloidosis without family history of FMF and with unknown precipitating disease, MEFV gene analysis is mandatory. Moreover, since FMF phenotype II is an actual hazard, a cost-benefit analysis suggests that MEFV mutation determination in all first-degree family members of FMF patients is warranted, as it will significantly reduce future patient treatment costs. PMID- 17111711 TI - [Changes in the transplantation world--from altruism to a utilitarian approach]. AB - The lack of organs for transplantation is a worldwide problem that has created a moral conflict between the traditional altruistic basis of organ donation and alternative solutions based on utilitarian grounds. Survival of grafts achieved in recent decades after unrelated living-donor kidney transplantation between spouses is longer than with deceased donor transplantation. This experience justified the extension of kidney donation beyond the traditional close family relationships including: anonymous donors and paired exchange programs. However, unrelated donation of kidneys within altruistic norms could not provide an ultimate solution for the lack of organs for transplantation. On the other hand, globalization and development of advanced medical technology in developing countries that do not provide transplantation for all their citizens, created an opportunity for a worldwide flourish of transplant tourism as an alternative solution for transplant candidates. Transplant tourism functions according to market laws and is profit-driven, as opposed to the legal organ exchange programs in Europe and the U.S.A., which are non-profit and patient-oriented. The transition from trade in kidneys from unrelated living-donors to the use of other organs (heart, lung and liver) from death penalty prisoners in China was only a matter of deciding where to lay the moral border when justifying the act for the sake of life-saving. Considering the inability of current legal altruistic transplantation practice to supply the growing need for organs, healthcare authorities and professional transplantation organizations have to tackle the donor crisis by designing legally acceptable utilitarian solutions. For instance, through the formation of international organ exchange programs under formal agreements or, in the case of kidney transplantation, through the establishment of a regulated compensated donation system. PMID- 17111712 TI - [Organ transplantation using organs taken from executed prisoners in China--a call for the cessation of Israeli participation in the process]. AB - Organ transplantation using organs taken from prisoners convicted to death is being practiced in China for the last two decades. These transplantations, which harshly violate medical ethics and international conventions, have become a prosperous market in China. This review presents details of the testimonies of a former prisoner and a Chinese physician who fled to the U.S.A., portraying the horrifying process of organ harvesting from executed prisoners, sometimes even before they are even dead. For years the Chinese authorities have denied the existence of this process, but following massive international diplomatic pressure they have recently publicly admitted to it for the first time. Many Israeli transplant candidates, who are desperate due to the organ shortage in Israel, seek their way to China in attempts to try and save their lives. The fact that they are currently being funded by all medical insurance companies constitutes a sort of formal approval to this atrocity. We, as a society, should stop this funding immediately, and divert all our human and financial resources to increase organ donation from brain-dead donors in Israel. PMID- 17111713 TI - [Domestic violence against women and the role of the obstetrician/gynecologist]. AB - Domestic violence against women is a worldwide phenomenon affecting women of all age groups and socio-economic backgrounds. It may take the form of mental, as well as physical or sexual abuse. Pregnant women are not excluded from being abused. The physical and mental health of women who suffer domestic violence is compromised in comparison with women who are not subjected to violence. These women are reluctant to report their problem to physicians and they, in turn, do not sufficiently investigate the possibility of exposure to domestic violence with their patients. In Israel there is comprehensive protective legislation concerning domestic violence against women and a developed welfare system managed by social workers. Nevertheless, the efforts to protect and help these women will remain futile without early recognition, identification and referral of such women by the attending physicians. Among physicians, the obstetrician/gynecologists have a unique role, since on many occasions they serve as the primary care physicians of women who suffer from domestic violence. They are therefore able to recognize and offer help to these women. PMID- 17111714 TI - [Epilepsy surgery]. AB - Epilepsy consists of recurrent seizures, which are not caused by reversible or transient metabolic or toxic etiologies. Approximately a third of the patients have medical refractory epilepsy and experience a decreased quality of life. In addition, these patients are subjected to increased risks of physical injury and death. Epilepsy surgery may be a therapeutic option in some of the patients with refractory epilepsy. The surgical outcome depends on optimal selection of the surgical candidates, the type of surgical procedure, and early surgical intervention during the course of the disease. PMID- 17111715 TI - [Management of prostate cancer with indolent biological potential: from watchful waiting to active surveillance]. AB - Prostate cancer is the most common malignancy among elderly men. Due to its indolent course and the fact that the majority of contemporary patients are diagnosed early, disease progression to metastasis often occurs many years after the initial diagnosis. Elderly men who have concurrent severe illnesses may not experience progression to metastasis during their lifetime. Therefore, it is reasonable to withhold active therapy in some patients avoiding the associated risks, and impact on patient quality of life. Watchful waiting (WW) is an adequate approach in such cases, and has been shown to be associated with a similar overall survival when compared with radical prostatectomy. However, the disease-specific survival was better in patients who had undergone surgery. Prostate cancer patients often find it hard to persist on a WW policy. This is due to their expectation to be treated, to fight, and win the battle over cancer. Consequently, many patients who start on WW drop out and seek active treatment within several years, mostly when PSA elevation is noted. Active surveillance for prostate cancer is a novel approach consisting of avoiding the risks of therapy, while allowing the early detection of those who are prone to progress. In these high-risk individuals delayed active treatment is offered. Active surveillance consists of periodic monitoring of the PSA serum level, digital rectal exam and repeated prostate biopsies. Threshold values for these parameters are pre-defined and active therapy is instituted when such threshold values are met. Preliminary results of active surveillance show that more patients remain on a surveillance regimen as compared with the traditional WW; it is possible to offer curative treatment to individuals who were defined as having high risk by active surveillance parameters, and the disease-specific mortality is similar to that with WW. Active surveillance for prostate cancer is a novel and fascinating approach that may improve our ability to distinguish between patients who have a higher risk and need active therapy, and others in whom the risk for progression remains low and avoiding the risks of therapy is warranted. PMID- 17111716 TI - [Dietary management of diabetic pregnancy]. AB - During pregnancy, several metabolic changes are observed which aim to provide optimum substrate, energy and other nutritional requirements to both the mother and the fetus. Maternal nutrition is the only source for most nutrients, influencing neonatal and placenta development, mother's physiological adjustment and also playing a major role in the destiny of the offspring. Over-nutrition or malnutrition are both linked with increased risk of diabetes mellitus in the offspring. Diabetes in pregnancy is the most common and important metabolic dysfunction in pregnancy. This is divided into two types and it is very important to distinguish between them, as each has different nutritional requirements and a different impact on the course of the pregnancy and the development of the fetus. Gestational diabetes mellitus (GDM) is the main type of diabetes in pregnancy, it usually appears in the second half of pregnancy and mainly influences fetal growth rate and can slow systemic development. Most women with GDM are treated with nutritional management alone. Pre-existing diabetes mellitus is present before pregnancy and it's effects begin at fertilization and implantation, and continue throughout pregnancy and thereafter. It involves high risk of early abortion, severe congenital defects and disrupted organogenesis. Although the treatment of pre-existing diabetes is usually a pharmacological one (insulin or oral pharmacological agents), nutritional management is still very important in normalization of glucose levels before and throughout the pregnancy. Fetal morbidity is lower in women with diabetes in pregnancy when optimal glucose control is maintained. Normalization of glucose levels during pregnancy is agreed to be the main factor in preventing poor outcomes in pregnancy. Dietary advice throughout pregnancy include frequent small meals which contain carbohydrates that are not highly processed, rich with slowly absorbed starches and non-soluble polysaccharides and with a low glycaemic index. The recommended daily caloric intake is individually suited to every woman. The main goals of nutritional management are to maintain balanced glucose levels and to provide enough energy and nutrients for all pregnant women, while avoiding ketosis, and minimizing the risk of hypoglycemia (in women treated with insulin). Health care providers should use the window of opportunity of pregnancy to change dietary patterns and to replace them with a healthy lifestyle for both the mother and her family. PMID- 17111717 TI - [The foundation of the first civil rehabilitation center by the Hadassah organization during the War of Independence]. AB - Following the War of Independence and the establishment of the State of Israel (November 1947 July 1949), there were large numbers of wounded and disabled soldiers and civilians. This situation forced the State of Israel to develop temporary rehabilitation centers. Rehabilitation centers for disabled patients to attend following hospital release were non-existent at the time in civilian hospitals. In August 29, 1948 the first civilian rehabilitation center was founded by the Hadassah Medical Organization in Jerusalem. By the end of the war, the center was closed due to lack of financial support and political reasons. This article describes the establishment of this center by the Hadassah Medical Organization during the War of Independence. PMID- 17111718 TI - Concurrent therapy (long acting beta agonists and inhaled corticosteroids) in the management of asthma. AB - BACKGROUND: Asthma is a clinical syndrome characterised by chronic inflammation of the lower respiratory tract in which many cells and cellular elements play a role, in particular mast cells, eosinophils, T-lymphocytes, macrophages, neutrophils and epithelial cells. Patients often require long-term anti inflammatory and reliever drugs to achieve a normal life. This review aims to highlight role of concurrent therapy in the optimal management of asthma. METHOD: A review of relevant literature was conducted using available medical journals and Science direct via the Internet. The key words employed were: asthma, concurrent therapy, long acting beta agonists and corticosteroids. British Thoracic Society and The National Heart, Lung and Blood Institute websites were also used in sourcing information for this review. RESULTS: Several studies support adding long acting beta agonists (LABA) to inhaled corticosteroids (ICS) than doubling the dose of ICS. This improves lung function, symptoms control and allows the dose of each drug to be adjusted to the patients'needs. CONCLUSION: This review was able to show that concurrent use LABA and ICS in asthmatics helps in adjusting their treatment within limits hence achieving control of the condition with minimal side effects. PMID- 17111719 TI - Current concepts in contraception. AB - BACKGROUND: Worldwide, contraceptive use has increased substantially over the past two decades. The increased demand for wider choices of contraceptive methods has resulted in extensive research and rigorous clinical trials. This has led to improvements on existing contraceptive methods and also the development of several new, more effective and acceptable methods with fewer side effects. Thus, this article presents a review of existing literature on recent developments on existing contraceptive methods. It also reviews recently developed contraceptive methods currently in use worldwide. METHODS: Relevant literature was reviewed using manual library search, electronic sources such as CD-ROMS and internet articles. CONCLUSION: More effective methods of contraception which are generally safer and easier to administer are increasingly being developed. Hopefully, as they increasingly become available in our environment, they will lead to and increase in acceptance and use of contraception by our women. PMID- 17111720 TI - Research proposal writing: breaking the myth. AB - BACKGROUND: Research is an essential activity required for the advancement of science and improvement of human existence. To carry out a research, a proposal is mandatory. However, in spite of the widespread demand for research proposals, experience has shown that a number of them are so poorly written that they rejected by assessors. This article aims at assisting researchers develop acceptable research proposals by reviewing the different components of a research proposal. METHOD: A review of relevant literature on research proposal writing sourced from manual library and internet search was used for this review RESULT: A research proposal is a formal and detailed statement of intent to carry out a research. It presents and justifies a plan of action and shows how the investigator thinks. A research protocol on the other hand is a plan written to seek approval for research from a supervisor or an organization. It is developed as a guide for a study and helps to keep the researcher focused on the topic and scope of the research. A research proposal has the following components: The Title page; The Abstract/Executive Summary; The Introduction/Statement of the Problem; Literature Review; Information on the applicant's centre; The Objectives/Research Questions/Hypotheses; The Study Design; Methods; Plans for Analysis; Data Analysis; Plans for Data Interpretation; Plans to report. Thus, although proposals and protocols are sometimes used interchangeably, a proposal precedes a protocol. CONCLUSION: To reduce the time wastage and frustration faced by intending researchers and their assessors it is essential that good proposals be written at all times. PMID- 17111721 TI - Biosocial characteristics and mode of delivery at term of women monitored in a voluntary agency hospital. AB - BACKGROUND: Many deliveries and maternal deaths occur in peripheral facilities and are usually attended to by General Practitioners (GPs). Maternal features that could help the GP predict whether a pregnancy will end in normal delivery or whether skilled assistance will be required to deliver the woman at term would be useful guides for early referrals or interventions. METHODS: This was a prospective study of 400 mothers with singleton pregnancies monitored at the Sacred Heart Hospital, Lantoro, Abeokuta, from February to July 1999. Records of maternal age, height, parity, occupation, educational level, weight-gain during pregnancy, the regularity of antenatal visits and delivery were analyzed. RESULTS: Mothers shorter than 1.55m, those who recorded large weight-gains during pregnancy, and those who booked late were more likely to experience difficult deliveries at term. Highly educated mothers and those in the upper socio-economic class were also more likely to have difficult delivery and require skilled assistance before birth at term. CONCLUSION: Some maternal biological and social characteristics can be used ante-natally by GPs to predict the type of delivery a woman will undergo at term. GPs should routinely look for such features in pregnant mothers. PMID- 17111722 TI - Childbirth in Germany and Nigeria compared. AB - BACKGROUND: A look at childbirth processes in both countries may lead to understanding of the differences between them and may perhaps open up new arrears of research in human reproduction. The objective of this paper is to compare some childbirth parameters in Nigerians and Germans and to adduce possible reasons for the differences. METHOD: This is prospective study using data collected from 1055 consecutive deliveries that took place in Nigeria which was analyzed and compared with that of 56690 deliveries that took place in the German state of Hesse. RESULTS: Preterm labor was more common among Nigerians (12.1%) than in Germans (6.7%). There is no significant difference in the day and time of birth in the two countries (p >0.05). Precipitate labor occurred in 12.5% of Germans, as against none in Nigerians. Within 30 minutes, more than 90% of women in both countries complete the second stage of labor. Doctors attend to more deliveries in Germany (99.3%), as against 63.1% in Nigeria. Episiotomy was performed in 71% of Germans as against 49% Nigerians. Primary cesarean section rate in Germany was 8.5% and in Nigeria 5.6%. Retained placenta was more common in Germans (2.5%) than in Nigerians (1%). Sepsis was more common in Nigerians (1.3%) than in Germans (0.0%). Blood loss greater than 1000ml was more common in Nigerians (2.9%) than in Germans (0.6%). Within 24 hours of delivery 74.8% of Nigerians leave hospital as against 4.0% of Germans. CONCLUSION: It would appear that biological and socio-economic factors play differential roles in the final process of childbirth in different populations while meteorological cum cosmic factors appear to exact the same influence in the two populations. PMID- 17111723 TI - Seasonality and other variation trends in birth weights of 4000g and above: A preliminary report from the University of Port Harcourt Teaching Hospital, Nigeria. AB - BACKGROUND: Anecdotal evidence appears to suggest a progressive increase in the proportion of large babies born at the University of Port Harcourt Teaching Hospital (UPTH), Nigeria. The objective of this study is to examine retrospectively the profile of birth weights at the UPTHover a defined time frame. METHODS: The birth weights of all babies born in the hospital from January 1, 1990 to December 31, 1999 were analyzed with the focus on weights = 4000g. The ten-year period was elongated into 20 six-monthly observation points and the periodic fluctuations in the proportion of birth weights = 4000g smoothed by using exponentially weighted moving averages with a weighting factor of 0.18. Seasonal variations in rates of birthweight = 4000g were calculated using Ratchet Circular Scan Test for a short seasonal peak. Proportions were compared using chi square statistic. RESULTS: Fourteen thousand three hundred and seven birth weights were recorded; 7111 males and 7196 females. Nine hundred and four (12.7%) and 434 (6.0%) of males and females respectively were 4000g and above. There was a sustained increase in the proportion of the = 4000g category during the observation period although the slope was steeper among the males (slope with [95% Confidence Interval] for males = 0.4131 [0.3213 to 0.5050] and for females 0.1801 [0.05565 to 0.3045]. There was a significant 3-month peak August to October of males only of birth weight = 4000g, (3-month peak; August to October, 30.3% of events. Test Statistic 3.46, p < 0.005). CONCLUSION: There has been a steady increase in the proportion of babies of birth weight =4000g at the UPTH in the ten-year period studied. This trend could be potentially dangerous for both mother and baby; therefore the authors believe it would be a good idea to carry out a prospective investigation along similar lines but using a much broader study base. PMID- 17111724 TI - Perinatal mortality associated with eclampsia in Kaduna, Northern Nigeria. AB - BACKGROUND: Eclampsia is an obstetric disorder with serious materno-fetal consequences. The objective of this study is to determine perinatal mortality rate associated with eclampsia in Kaduna Northern Nigeria. METHOD: A retrospective study of 27 perinatal deaths associated with 61 cases of eclampsia in the Ahmadu Bello University Teaching Hospital, Kaduna (ABUTHK) from 1st January 1990 to 31st December 2000 was done. Resource materials were the admission records, case files, delivery and neonatal case files of women and their babies. The records were reviewed for relevant bio-social, obstetric and neonatal data. RESULTS: Perinatal mortality rate associated with eclampsia was 409/1000 births. It was highest amongst women in the age group 35 years and above, in the two extremes of parity, in those not registered for care, in women with poor control of eclampsia fits and those with ante partum eclampsia.Perinatal mortality rate decreased as gestational age rose. The perinatal mortality rate for those delivered vaginally was 706/1000, and 150/1000 for those delivered abdominally with a (P<0.05). CONCLUSION: Perinatal mortality rate associated with eclampsia is very high. Making prenatal care available to all, improving the quality of care, early resort to caesarean section and use of perimorterm caesarean section in carefully selected women may reduce the deaths. PMID- 17111725 TI - Blood transfusion therapy in neonates admitted into the Special Care Baby Unit (SCBU) of University of Port Harcourt Teaching Hospital, Port Harcourt. AB - BACKGROUND: Blood transfusion is associated with potential risks and adverse effects; it is therefore pertinent to ensure that it is given only when it is indicated. The objective of this study is to determine the rate and the indications of blood transfusion in neonates admitted into the Special Care Baby Unit (SCBU) of University of Port Harcourt Teaching Hospital (UPTH), Port Harcourt. METHOD: A prospective study of Neonates admitted into SCBU between January 1st 2003 and December 31st 2004 and who had blood transfusion during their hospitalization was carried out. RESULTS: Preterm babies are more likely to be transfused and are also more likely to receive multiple blood transfusions. Severe neonatal jaundice and severe anaemia are the commonest indication for blood transfusions in the neonates. Exchange blood transfusions (EBTs) were utilized more often than top up transfusions even among preterms. Adverse events were seen more in those that had EBT CONCLUSION: The rate of blood transfusion is still very high among neonates. Concerted efforts should be made to prevent severe neonatal jaundice and severe anaemia and thus reduce the rate of blood transfusion. PMID- 17111726 TI - Risk factors associated with sexual assault in Calabar south eastern Nigeria. AB - BACKGROUND: Sexual assault inflicts significant physical and psychological trauma in the victims. Interventions to prevent this violent crime against women are a major public health concern. The aim of this study is to identify the risk factors for sexual assault as seen in victims presenting in our hospital. METHOD: Twenty-two case records of sexual assault victims treated at the University of Calabar Teaching hospital were reviewed for this study. RESULTS: The incidence of reported sexual assault in this study was 2.1% with a rising trend observed. Age range was from 4 to 23 years. Six (27.3%) victims were primary school pupils. Twelve (54.5%) victims had not attained menarche, and 20 (90.9%) cases were single. Recorded place of assault was commonly along a bush track (36.4%) and a nearby cemetery (22.7%). Time of assault was mainly in the evening (50.0%). Identity of the assailant was known in 7 (31.8%) cases. Commonest weapon used by the assailant was a knife (36.4%). CONCLUSION: Concerted effort is required to curb the rising trend of reported sexual assault. Interventions aimed at creating public awareness of the possible risk factors may reduce the incidence of this detestable event in the community. PMID- 17111727 TI - KAP of HIV prevention and screening among pregnant women attending specialist antenatal clinics in Calabar, Nigeria. AB - BACKGROUND: There is a growing concern globally to reverse the growing incidence of HIV especially in Sub-Saharan Africa. This study was conducted to determine the level of awareness, attitude and practice of antenatal HIV screening in Calabar. METHOD: Descriptive multi-centric study of 400 antenatal attendees in Calabar, carried out in October 2005 using pre-tested, semi-structured and interviewer-administered questionnaire for data collection. RESULT: Of the 96.7% women with knowledge of HIV infection, 41.2% were assessed to have excellent knowledge of the mode of transmission. Awareness of antenatal HIV screening was observed in 96.2% women; while 93.7% approved of antenatal HIV screening. Awareness and approval of antenatal HIV screening was significantly related to age and educational status. The proportion of women who had HIV test in current pregnancy was 70.2%. Pre-test counseling was done in 65.8% of women. Most women (78.2%) who had not been screened were willing to undergo HIV testing. Spousal disapproval (23.1%) was the main reason for unwillingness to undergo HIV testing. CONCLUSION: The study revealed high levels of awareness, approval and practice of antenatal HIV screening. However, pre test counseling was not consistently given in cases tested. PMID- 17111728 TI - Perception of bed nets and malaria prevention amongst users of insecticide treated bed net in a semi-urban community in south-south Nigeria. AB - BACKGROUND: Assessment of the use and success of insecticide treated nets (ITN) in malaria control requires a good insight into the people's perceptions of malaria and ITN. This study reports on the perception of mothers of malaria, malaria prevention, and ITN, before and one year after the ITNs was bought from a social marketing programme. METHODS: The study was an intervention study carried out using an interviewer-administered questionnaire. The respondents were mothers who bought ITN sold by a social marketing project in Egbema a semi-urban community in Rivers State. The pre-intervention study was carried out at the time of purchase of the nets, while the post-intervention study was done one year later. RESULTS: There was a significant increase in the proportion of respondents who mentioned mosquito as the sole cause of malaria. (P < 0.001). There was also a significant reduction in the malaria burden of the household (P < 0.001). Most of the respondents (84.8%) stated that they liked the ITN because it ensures a good night sleep, free from the nuisance of mosquitoes. However, 48.31% complained of heat, and 26.40% complained of the task of having to mount the net every night for their children. CONCLUSION: Sustained health education and use of ITN can promote the use of the nets for malaria control. PMID- 17111729 TI - Tumour markers and hepatitis C virus infection in Nigerian patients with liver diseases. AB - BACKGROUND: Early diagnosis of hepatocellular carcinoma (HCC) is very important and determination of serum levels of tumour markers in patients with chronic liver diseases could be of immense contribution to their management. METHOD: Forty-two adult Nigerian subjects consisting of 14 healthy subjects (Controls) and 28 patients with primary liver mass (es) and histological diagnoses of liver cirrhosis (LC) +/- chronic active hepatitis (CAH) and hepatocellular carcinoma +/ LC were studied. Their blood samples were assayed for Hepatitis B Surface Antigen (HBsAg), antibodies to HCV (anti-HCV), alkaline phosphatase (AP), Aphafeotoprotein (AFP) and Ferritin. RESULTS: The patients had HCC (10) HCC+LC(4), LC+CAH (2) and LC (12). Serum ferritin> 700ng/ml, AP> 375IU/ml and AFP>200IU/ml were detected in 32%, 11% and 32% of the patients respectively with corresponding specificities of 100%, 86% and 100%. Elevated serum levels of AFP and ferritin were found in patients with HCC +/- LC while raised serum AP occurred in those having PHCC without LC. Only combination of either AFP or ferritin to AP gave significant increase in the diagnostic yield of HCC among the patients than the use of only AR Elevated levels of serum AFP correlated with both HBV and HCV while raised serum levels of ferritin were associated with only with HBV CONCLUSION: Although combination of the tumour markers gave a higher diagnostic yield for HCC among Nigerian patients, serum AFP > 200IU/ml seems the best tumour marker in the diagnosis of PHCC among the patients. PMID- 17111730 TI - The appearances of benign breast diseases on ultrasound. AB - BACKGROUND: Benign breast diseases are common and Ultrasound is a very useful tool in examining benign breast diseases especially in distinguishing solid from cystic masses. This study aims to determine the sonographic pattern of benign breast diseases in a Nigerian population and to compare this pattern with that which has been previously reported in other environments. METHODS: This a prospective study of Ninety-four patients with breast disease scanned between 1997 and 2001 on request from the breast clinic of the Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife. A total of forty-four patients with histologicaly confirmed benign breast diseases were selected for this study. Their ultrasonographic features were evaluated and compared with those previously described. RESULT: Fibroadenoma was the commonest disease, occurring in 62.2% of the patients, with a mean age of 29.1 years, an age higher than that previously reported. Other diseases which were expected to occur commonly (breast abscess, cyst) were rarely seen. CONCLUSION: While it was discovered that the sonographic pattern of most of the diseases in this study conformed to that already described in literature despite late presentation, only one sonographic appearance of galactocele, out of the three reported, was noted in this study. In addition, despite the nonspecificity of ultrasound appearances in fibrocystic disease, an attempt has been made to match these appearances with the different histological types already described in literature. PMID- 17111731 TI - Socio-demographic variables and family health: A prospective study of a Katcha in north-central Nigeria. AB - BACKGROUND: Healthy families are vital prerequisites for a stable society and economic development of the community. The health status of families and communities is influenced by several socio-demographic variables such as educational status, marital pattern and gender relationships. The objective of this study is to examine the effect of certain socio-demographic variables on the health status of a rural community in Northern Nigeria. METHOD: A prospective survey over a six month period, which commenced in May 2005, was done in rural primary health care centre in Katcha local Government area of Niger state. All consecutive parents either male or female of children seeking care in the health center who gave consent to participate in the survey were recruited. A structured researcher administered questionnaire was used in sourcing data. Data was analyzed using Microsoft Excel version 2003. RESULTS: A total of 608 parents comprising 302 (49.67%) male and 306 (50.33%) females (M: F = 1:1.01) were surveyed. Of these 78.48% females were uneducated compared to only 41.83% males. While most males were gainfully employed, 20.86% of females were full time housewives. Polygamy was the predominant marital pattern. The findings indicate that women in the community were socially disadvantaged compared to males. CONCLUSION: There is a significant socio-economic gap which puts females at a disadvantage in the rural community surveyed. Bridging this Socio-economic gap between men and women in rural communities will help improve the health status in our rural communities. PMID- 17111732 TI - Internal hernia through the broad ligament presenting with acute on chronic intestinal obstruction: A case report. AB - BACKGROUND: Internal hernias are rare. Their occurrence through the broad ligament is one of the rarest forms. We present a report of a 45 year Nigerian female with acute on chronic intestinal obstruction due to an internal hernia of the broad ligament. METHODS: Patients case notes and a review of relevant literature using manual library and Medline search was used. RESULTS: A 45-year old multiparous woman presented with a ten-year history of features of partial intestinal obstruction, and no previous abdominal surgery. Examination revealed right iliac fossa tenderness and a plain abdominal radiograph showed air fluid levels. Conservative management was not successful and laparatomy confirmed an internal hernia of the right broad ligament which was repaired with satisfactory outcome. CONCLUSION: This report is to highlight the fact that though rare, internal hernia should be considered in the differential diagnosis of intestinal obstruction. PMID- 17111733 TI - Slow, gradual external fixator distraction in acquired ankle and foot contracture. AB - BACKGROUND: External fixators have been noted to have a place in the orthopaedic management of problems involving the ankle and foot. We here report a case of ankle and foot contracture managed by soft tissue release and slow, gradual external fixator distraction. METHOD: A case report of a patient with acquired ankle and foot contracture and discussion of relevant literature. RESULTS: A 9 year old female presented to our out-patient clinic with features of right ankle and foot contracture following treatment by traditional bone setters 6 years earlier. The contracture was fixed at 30 degrees. She had soft tissue release and slow, gradual external fixator distraction which corrected the foot to a plantigrade position. Subsequently she had skin grafting for the skin defect. After removal of the external fixator she was placed on a below knee cast and commenced weight bearing. The cast was removed after three weeks and the patient has continued to bear weight on a plantigrade foot. CONCLUSION: External fixators have a definite place in contracture release and should be widely utilized. PMID- 17111734 TI - Sylvian cleft meningioma: surgical approach and postoperative morbidity. AB - BACKGROUND: Meningiomas without dural attachments are rare, and are commonly diagnosed postoperatively. We here report a case of meningioma and examine the surgical options. METHOD: We reviewed the case record of the patient who presented with a right sylvian cleft meningioma as well as relevant literature on the subject. RESULT: Brain CT scan performed on a 73-year-old woman on admission for non-specific symptoms revealed. a heterodense temporoparietal mass which was demonstrated on carotid angiography as being fed by the middle cerebral artery. Preoperatively, a glioma was considered as being most probable because of its radiological features. The mass, which at surgery was found to be located in the sylvian fissure, was however histologically confirmed to be a meningotheliomatous meningioma with fibroblastic component CONCLUSION: The surgical approach to meningioma of the sylvian cleft is a prime determinant of outcome following tumour resection. Making an appropriate approach largely depends on making a correct preoperative diagnosis for which a high index of suspicion is necessary. PMID- 17111735 TI - Congenital vaginal fistula from a single system ectopic ureter: A case report. AB - BACKGROUND: Vaginal fistulae particularly vesico-vagina fistula, is a common urogynaecologic problem in Sub-Saharan Africa. Majority of these cases are acquired and congenital fistulas are rare. We here report a case of congenital vaginal fistula secondary to an ectopic ureter in 15 year old Nigerian female. METHOD: The case records of a 15 year old female who presented to the obstetrics and gynaecological unit of the Federal Medical centre Azare and a Review of literature on the subject was used. RESULT: A 15-old-girl presented with continuous leakage of urine from the vaginal since birth despite the establishment of normal voiding habit. Examination revealed urine leaking from a pinhole fistula in the vestibule. Intravenous urography showed delayed excretion of the right kidney with a normal single ureter (single system) and normal left kidney. Ureteric implantation into the bladder was performed with good postoperative outcome. CONCLUSION: In an environment with scarce resources, managing ectopic ureter in the female is quiet challenging. Multidisciplinary approach involving the urologist, paediatric surgeon, radiologist and the gynaecologist will yield the best outcome. PMID- 17111736 TI - Kartagener syndrome: an unusual cause of respiratory distress in the newborn. AB - BACKGROUND: Kartagener Syndrome (KS) a rare genetic disorder belongs to a group of disorders referred to as primary ciliary dyskinesia (PCD) where the cilia covering the respiratory epithelium is either immotile or beat in an uncoordinated fashion. It is characterized by a triad of dextrocardia (with or without situs inversus), chronic sinusitis and bronchiectasis as a result of poor mucociliary clearance of mucus and bacteria. This may lead to respiratory distress in the newborn period. METHOD: This is a case report of a 14-day old male who presented with respiratory distress (which was noticed soon afterbirth) and features suggestive of KS. CONCLUSION: KS should be suspected in neonates presenting with respiratory distress, pneumonia and no risk factors for infection. PMID- 17111737 TI - Confirmed congenital rubella syndrome--A case report. AB - BACKGROUND: Congenital Rubella Syndrome (CRS) is defined clinically as an illness usually manifesting in infancy, resulting from rubella infection in utero with certain specific signs and symptoms. Confirmed CRS is a clinically consistent case which is laboratory confirmed. A report of a 3 month old male diagnosed with confirmed congenital rubella is here presented on account of its rarity and easy prevention with vaccination. METHOD: The case notes of 3 month old male admitted with features consistent with CRS and managed for overwhelming septicaemia with heart failure and encephalitis and review of literature on the subject using manual library and Medline search. RESULT: The infant's Rubella-specific immunoglobulin M (IgM) antibody demonstrated positive and the rubella antibody level was high at 11.6 IU/ml. CONCLUSION: This is the first report of confirmed CRS in our centre. Though few cases are seen, the effects and defects on the child are severe and irreversible thus, vaccination with Measles-mump-rubella (MMR) vaccine is recommended as part of the National Programme on Immunization and for all females of child bearing age, who did not receive it in childhood. PMID- 17111738 TI - Addison's disease presenting as acute chest syndrome: case report and review of literature. AB - BACKGROUND: Addison's disease is due to primary adrenal failure. It is an uncommon condition with equal prevalence in both males and females. The onset of symptoms is gradual and manifestation is non specific, hence diagnosis is easily missed without a high index of suspicion. METHODS: The medical records of a patient who presented with acute chest pain to the cardiac unit of the University of Port Harcourt Teaching Hospital were reviewed. A review of the literature using manual library and Medline search on Addison's disease was also done. RESULT: A 48 years old male presented in our medical outpatient department with a three day history of sudden onset of severe precordial chest pain that started while playing football which was associated with nausea, vomiting and difficulty in breathing. After initial clinical evaluation a diagnosis of acute myocardial infarction and cardiac failure with a suspicion of background Addison's disease was made. Serial electrocardiography done over a two week period did not show evidence of myocardial infarction, but the patient had elevated serum ACTH and very low serum cortisol levels. An abdominal CT scan done two weeks after admission showed absence of the Adrenal glands bilaterally, confirming Addison's disease. He received treatment for cardiac failure, analgesics, prednisolone and a mineralocorticoid to which he responded satisfactorily and has remained healthy. He also received a six months course of antituberculous treatment empirically. CONCLUSION: Addison's disease is an uncommon endocrine disorder which can present insidiously in a non specific manner. Diagnosis requires a high index of suspicion. PMID- 17111739 TI - Tubal ectopic pregnancy after bilateral tubal ligation: A case report. AB - BACKGROUND: Tubal ligation is a common method of contraception, and pregnancy after this method of sterilization is uncommon. We here present a report of Tubal pregnancy after a Bilateral Tubal Ligation (BTL). METHOD: The case notes of a 35 year Nigerian female who presented with a tubal pregnancy after BTL and a review of literature on the subject was used. RESULT: A 35-year-old para 4=0 had bilateral tubal ligation during caesarean section for her last childbirth. She presented 3 years later with a six weeks history of irregular vaginal bleeding and lower abdominal pain and had a laparotomy for a right tubal ectopic pregnancy. CONCLUSION: Ectopic pregnancy after bilateral tubal ligation is uncommon. Females who undergo BTL should be adequately counseled on the possibility of failure of this procedure for contraception. PMID- 17111740 TI - Spirochaetemia in a HIV positive patient. AB - BACKGROUND: Borreliosis, caused by Borrelia recurrentis and several other Borrelia species is not a commonly reported case in our environment, but the search for the cause of recurrent pyrexia in this patient made it possible to discover the spirochete as the cause of the disease. METHOD: A 38 year old married HIV positive woman presented with recurrent fever in a private clinic. Six thin smears were made out of the patient serum and dried in the air. Three slides each were stained with 0.12% Leishman and 20% Giemsa stains and examined under the light microscope. RESULT: Three of the Giemsa slides were positive for spirochetes (4-5 spirals), which were constituents with Borrelia species. The patient responded very well to tetracycline and serum became negative for the organism after ten days of treatment. CONCLUSION: Borrelia was discovered to be the cause of the recurrent pyrexia in this patient who responded very well to tetracycline. Borrelia should be looked for in cases of pyrexia of unknown origin PMID- 17111741 TI - Physical exercise and health: a review. AB - BACKGROUND: Physical activity results in increased exercise capacity and physical fitness, which may lead to many health benefits. Individuals who are more physically active appear to have lower rates of all-cause mortality, probably due to a decrease in chronic diseases including coronary artery disease (CAD). This may result from an improvement in cardiovascular risk factors in addition to enhanced fibrinolysis, improved endothelial function, decreased sympathetic tone, and other yet undetermined factors. METHODS: We reviewed the literature on physical activity and health with particular reference to the benefits derivable by engaging in regular physical activity. The MEDLINE/PUBMED and bibliographic searches for English language studies were used. RESULTS: Physical inactivity is now considered a risk factor for Cardiovascular diseases (CVD). Regular exercise results in an increase in exercise capacity and lower myocardial oxygen demand leading to cardiovascular benefits, including lower mortality rates. Physically active individuals suffer from fewer ailments than do less-active individuals. Physical activity reduces cardiovascular risk through lowering of blood pressure, improved glucose tolerance, reduced obesity, improvement in lipid profile, enhanced fibrinolysis, improved endothelial function and enhanced parasympathetic autonomic tone. CONCLUSION: Physical exercise has many health benefits and the evidence for this continues to accumulate. Health care professionals should incorporate counselling to patients for physical exercise in their daily clinical practice, while health policy makers and community physicians should see to implementation of this at the community level. PMID- 17111742 TI - Current concepts in the management of refractory cirrhotic ascites. AB - BACKGROUND: Ascites is the pathologic accumulation of fluid within the peritoneal cavity. This condition when refractory to treatment heralds more severe complications and a poor prognosis. The aim of this paper is to review literature on the pathogenesis and current management of refractory cirrhotic ascites. METHODS: An English language literature search using Medline and PubMed (1976 2006 March) was done to assess all research/review articles on the pathogenesis and management of refractory cirrhotic ascites. RESULTS: The onset of ascites in cirrhotic patients signifies poor prognosis. Only a small percentage of patients with cirrhotic ascites develop true resistance to diuretics. Serial therapeutic paracentesis remains the available option for the majority of patients. Liver transplantation is the only definitive therapy, but the use of this method is limited by the availability of the organ and the cost of such procedure. Transjugular intrahepatic portosystemic stent shunt is a useful procedure but limited by the complication of disseminated intravascular coagulopathy. The role of other alternatives is less clearly defined. CONCLUSION: Therapeutic options in patient with cirrhotic ascites remains serial therapeutic paracentesis with or without plasma expansion. Transjugular intrahepatic portosystemic stent-shunt is a useful alternative. The unavailability of liver donors serves as a barrier to liver transplantation. PMID- 17111743 TI - Anaemia in human immunodeficiency virus infection: a review. AB - BACKGROUND: Human immunodeficiency virus infection is associated with myriads of haematological abnormalities and complications, including anaemia. This review aims at presenting the epidemiologic evidence of HIV associated anaemia, and also examine its effects on patients' survival and the need for specific diagnosis and treatment. METHODS: A review of relevant literature on the subject was sourced manually and by PUBMED internet search. The following keywords were used for internet search: anaemia, chronic disease, highly active antiretroviral therapy human immunodeficiency virus, haematological abnormalities, and AIDS. RESULTS: Human immunodeficiency virus (HIV) is associated with numerous abnormalities of red blood cells production and lifespan. One of these consequences is anaemia. The prevalence of these estimates varies widely from one population to another; however, anaemia was consistently shown to be a predictor for increased disease progression and decreased survival of patient infected by HIV. CONCLUSION: Regular evaluation of patients infected by HIV is necessary, to determine the specific causes of anaemia in order to ensure the institution of appropriate intervention. PMID- 17111744 TI - Type 2 diabetes mellitus: a review of pharmacological treatment. AB - BACKGROUND: Type 2 diabetes mellitus, is a disease with a rising prevalence worldwide. It is currently estimated that 190 million people around the world suffer from diabetes mellitus, with over 330 million predicted to have the condition by 2025 and 366 million by the year 2030. It is predicted that the developing countries will contribute 77.6% of the total number of diabetic patients in the world by the year 2030. This rapidly growing prevalence among developing countries is attributed to the effects of urbanization, industrialization and globalization on these countries. There has been substantial progress over the last decade in the development of new agents for the treatment of type 2 diabetes especially focusing on the underlying pathophysiology. Despite this and the numerous guidelines from diabetes organisations only less than 40% of patients achieve recommended glycaemic targets. We therefore decided to do a review of the pharmacological treatment of type 2 diabetes mellitus to highlight the pharmacology and effectiveness of these agents and their roles in the management of type 2 diabetes. METHODS: We reviewed the literature on the subject using materials from library search, articles in journals, internet search and conference abstracts. RESULTS: The global burden of type 2 diabetes mellitus, the various pharmacological agents available for the treatment of type 2 diabetes mellitus, including novel agents were discussed. CONCLUSION: The prevalence of type 2 diabetes mellitus is increasing worldwide and the predicted increase is much higher in developing countries compared to the developed countries. There are obviously an enormous number of therapies available for the treatment of type 2 diabetes mellitus and if effectively deployed it will be possible to achieve target diabetic control in most of our patients. This however, should not detract us from adopting measures that will reduce the prevalence of type 2 diabetes mellitus in our population bearing in mind that prevention is more cost effective especially given our low socioeconomic development and the very high predicted rise in the burden of type 2 diabetes mellitus in our developing world. PMID- 17111745 TI - Stroke in persons with diabetes mellitus in Jos, Nigeria. AB - BACKGROUND: Mortality and morbidity from stroke in patients with diabetes are on the increase. Epidemiological surveys are most crucial in planning and allocating resources as they provide critical information for good policy formulation in a resource poor country The purpose of this study was to determine the frequency of stroke in patients with diabetes mellitus attending the Medical out Patient unit of Jos University Teaching Hospital. METHODS: A total of one hundred and twenty Nigerians with diabetes mellitus were consecutively selected and age-sex matched with sixty non-diabetic controls in a ratio of two to one respectively. RESULTS: The mean age of the diabetics was 53.0 +/- 12.4 years while that of the controls was 54.5 +/- 13.9. The mean duration of DM was 8.4 +/- 6.9 years. Sixty five percent of the diabetics had their disease for up to five years. Eight (6.7%) of diabetic patients had ischaemic stroke while 3 (5%) of the controls had stroke. CONCLUSION: The frequency and risk factors for stroke in persons with diabetes mellitus in Jos, Nigeria are similar to those obtained in other parts of the world. A differing result of hyperlipidaemia as a major determinant for stroke in persons with diabetes mellitus is worth studying in future in a large prospective study. PMID- 17111746 TI - Indices of obesity derived from height and weight in a Nigerian adult population. AB - BACKGROUND: Obesity is a rapidly growing health risk all over the world. Even in mild degrees, it has serious adverse effects and is associated with diminished life span. The study was aimed at determining the most suitable obesity index derived from height and weight in a young adult Nigerian population. METHODS: A cross-sectional study of students of University of Nigeria Enugu Campus was done. Heights and weights of 402 males and 268 females aged between 20 and 28 were measured. Weight-height ratio (W/H), body mass index (W/H2), Rohrer's index (W/H3) and ponderal index (H/W(1/3)) were calculated. RESULTS: Zero-order correlation coefficients of these indices with height and weight showed that body mass index was the only index not significantly correlated with height in both sexes. Weight-height ratio and Rohrer's index underestimated the degree of obesity in short subjects and overestimated it in tall subjects. The reverse was the case for ponderal index. CONCLUSIONS: Body mass index (W/H2) is the most suitable index derived from height and weight for the assessment of obesity in our study population. We recommend its use in busy clinical practice and epidemiological studies. PMID- 17111747 TI - Treatment outcome of TB/HIV positive and TB/HIV negative patients on directly observed treatment, short course (DOTS) in Sagamu, Nigeria. AB - OBJECTIVE: The World Health Organisation marked 10 years of implementation of Directly Observed Treatment short course (DOTS). One key factor affecting the success of the DOTS strategy is the rising HIV prevalence among TB patients. The study was embarked upon to compare the treatment outcome of TB/HIV positive and TB/HIV negative patients in Sagamu, Nigeria. METHODS: A prospective study of 353 smear positive TB patients aged 15 years and above who were registered for 8 months anti-tuberculosis (DOTS) therapy between January 2001 and December 2003 at the Olabisi Onabanjo University Teaching Hospital, Sagamu, Nigeria. Treatment outcome indicators of cure, default, transfer to another district and death were assessed in relation to the HIV status of the patients. RESULTS: There were 353 eligible patients of which 58 (16.4%) were HIV positive. The clinical symptoms and signs of TB were similar in both HIV positive and negative TB patients. The cure rate was 76.8%.patients. The cure rate was significantly lower in HIV infected compared with non-HIV infected TB patients (60.3% v 80.0%;p = 0.0001). However, among survivors it was not significant (71.4% v 82.5%; p = 0.07). Overall mortality was 5.1% which was significantly higher in HIV positive compared to HIV negative TB patients (15.5% v 3.1%; p = 0.00007). On the whole, 17% defaulted treatment and 1.1% failed treatment. These were however not significantly related to HIV status. CONCLUSION: The cure rate in this study is still lower than the recommended 85% target by the WHO. Mortality rate in TB/HIV positive patients was higher than in HIV negative patients. The option of a community based TB programme using volunteers or family members to supervise administration of anti-TB drugs so as to ensure adherence to TB treatment may be considered. PMID- 17111748 TI - Human immunodeficiency virus and hepatitis B virus co-infection among patients in Kano Nigeria. AB - BACKGROUND: Human immunodeficiency virus and hepatitis B virus are widespread in the developing countries and patients with dual infection of HIV and HBV are increasingly being diagnosed among hospital patients. Reports have indicated that hepatitis will contribute significantly to morbidity and mortality in HIV infected patients because of increased use and accessibility of highly active antiretroviral therapy (HAART). The objective of this study is to determine the prevalence of HIV and HBV co-infection in patients in Kano Nigeria and to highlight the reciprocal interactions between the HIVand HBV. METHODS: Three hundred patients consisting of 152 males and 148 females were recruited into the study at the Aminu Kano Teaching Hospital Kano, Nigeria between February 2002 and March 2003. RESULTS: Out of a total of three hundred HIV positive patients, two hundred and eleven (70.3%) were HBV positive. Of the 152 males that are HIV positive, 102 (67.1%) were HBVpositive while out of 148 females that are HIVpositive, 109 (73.6%) were HBV positive. CONCLUSION: A co-infection rate of 70.5% was observed in this study. Since HIV infected patients with HBV co infection respond less to HAART additional concern and care must be taken in order to minimize the complications associated with the increasing use of HAART The testing of HIV positive patients for HBV will help in the choice of therapy in these patients's. PMID- 17111749 TI - Nosocomial and community acquired urinary tract infections at a teaching hospital in north central Nigeria: findings from a study of 12,458 urine samples. AB - BACKGROUND: Urinary tract infection (UTI) is still a problem not infrequently encountered in the course of clinical practice. This study was designed to ascertain its prevalence among both the in and out-patients and the antibiotic susceptibility pattern of the isolates. METHOD: A retrospective analysis of data from all the urine samples processed at Jos university teaching hospital microbiology laboratory was undertaken for a period of 36 months (January 2000 December 2002). Samples had been collected, stored and processed by standard laboratory procedures. Results obtained were analysed using SPSS 11 statistical software and P values < 0.05 were considered significant. RESULTS: Of the 12,458 urine samples studied comprising 43% males and 57% females: the overall prevalence of UTI was found to be 22%; 7.4% among males and a higher figure of 14.6% among females, this was statistically significant (p < 0.05). The rate of nosocomial UTI was significantly higher than the community acquired type: 12.3% and 9.3% respectively (p < 0.05). The commonest nosocomial isolate was Klebsiella spp while Escherichia coli were for community acquired group. Staphylococcus aureus, Coagulase negative Staphylococcus, Proteus spp and Pseudomonas aeruginosa were also common isolates. The most effective antibiotics were Ofloxacin, Ciprofloxacin and Cefuroxime. CONCLUSION: In-patients especially should be encouraged to drink adequate water daily and practice "double urination" to reduce incidence of UTL. Ofloxacin, Ciprofloxacin and Cefuroxime should be considered first in the treatment of UTI in the absence of a susceptibility test. PMID- 17111750 TI - Haematological values in pregnant women in Port Harcourt, Nigeria. AB - BACKGROUND: Conflicting results have been documented regarding the statistical significance of variations in some haematological parameters at different trimesters of pregnancy. Environmental factors, ethnic and tribal peculiarities have variously been implicated. The values of eight important haematological parameters: Haematocrit (Hct), Haemoglobin concentration (Hbc), Red Blood Cell (RBC) count, White Blood Cell (WBC) count, Mean Corpuscular Volume (MCV), Mean Corpuscular Haemoglobin (MCH), Mean Corpuscular Haemoglobin Concentration (MCHC) and Erythrocyte Sedimentation Rate (ESR) were therefore determined in healthy pregnant subjects receiving antenatal care at the University of Port Harcourt Teaching Hospital, Port Harcourt, Nigeria. METHOD: Cross-sectional prospective study involving 130 pregnant subjects aged between 18 and 43 years. The subjects were divided into three groups consisting of 46 subjects in the first trimester, 36 subjects in the second trimester and 48 subjects in the third trimester of pregnancy. The height, weight and body mass index of all subjects were also determined. RESULTS: Results obtained for the haematological parameters indicate that only Haematocrit (Hct) showed significant differences amongst the three groups; highest amongst subjects in the third trimester and lowest amongst subjects in the second trimester (p < 0.05). Haemoglobin concentration (Hbc), Mean Corpuscular Volume (MCV), Mean Corpuscular Haemoglobin (MCH), Mean Corpuscular Haemoglobin Concentration (MCHC), and Erythrocyte Sedimentation Rate (ESR) were found to be highest amongst subjects in the second trimester; Red Blood Cell (RBC) count and White Blood Cell (WBC) count were highest amongst subjects in the first trimester of pregnancy. These differences were however, not statistically significant (p > 0.05). Among the anthropometric parameters studied, only weight showed significant differences in the three groups of pregnant subjects (p < 0.05); being highest amongst subjects in the third trimester and lowest in subjects in the first trimester. CONCLUSION: The present study provides additional baseline data for basic haematological parameters in healthy pregnant Nigerian women. This would be of immense benefit especially in the antenatal assessment of pregnant Nigerian women. PMID- 17111751 TI - Demographic factors determining compliance to iron supplementation in pregnancy in Oyo State, Nigeria. AB - BACKGROUND: Iron deficiency anaemia in pregnancy has irreversible negative consequence on infant cognitive development and increases maternal morbidity and mortality. Iron supplementation ameliorates this effect. The salutary effect of iron supplementation on improvement of haemoglobin levels in pregnancy has been documented in various studies. However factors affecting the compliance of women with prescribed iron supplements are yet to be well studied, particularly at primary care levels. This study describes the compliance rate and demographic factors influencing use of iron supplements among pregnant women at primary care level. METHOD: This is a cross sectional descriptive study. Five hundred and ninety seven women attending randomly selected primary care centres were studied. Demographic characteristics and information on use of iron supplements were collected. Haemoglobin level was also determined for each woman. RESULTS: Compliance rate was 37.5%. Prevalence of anaemia was higher among noncompliant women than those complying (18% v 15%, chi2 7.5, p = 0.006). Haemoglobin level was higher among women complying with iron supplements compared with those not complying (11.4 g/dl v 11.0 g/dl, t = 9.3, p = 0.002). Single and teenage mothers and those aged 35 years and above were less likely to be compliant. Married women, those in urban location, and those aged 20-29 years were more compliant with iron supplementation. CONCLUSIONS: Pregnancy among teenage and single mothers is associated with a significantly higher risk of non-compliance with iron supplementation. Further studies are recommended to understand the reasons for non-compliance among these groups of women. PMID- 17111752 TI - The knowledge, attitude and practice of voluntary counselling and testing (VCT) for HIV/AIDS among undergraduates in a polytechnic in southeast, Nigeria. AB - BACKGROUND: The identification of infected individuals is an important step in the control of the HIV epidemic. Voluntary counselling and testing (VCT) for HIV/AIDS is a concept developed in this direction. The objective of this study is to determine the level of knowledge, attitude and practice of Voluntary Counselling and Testing (VCT) for HIV/AIDS among undergraduates in a Nigerian tertiary institution. METHOD: This is a cross sectional study using a multistage sampling method to enroll students from different levels of the national diploma programme into the study. A structured questionnaire was administered to the 260 students with response rate of 70%. RESULT: Only 115 (63.2%) of the students were aware of VCT with 68 (59.1%) having heard of it at least one year prior to the study. Mass media and Churches were the highest sources of information on VCT Most of the students did not know where VCT services could be obtained and knowledge of what VCT entails was also low. However, 127 (69.8%) students approved the necessity of counselling prior to testing and 117 (64.3%) were ready to take a positive result in good fate. At least one out of every four students (54 of 182) had been sexually active within three months preceding the study Only 48 (26.4%) students had taken an HIV test at one time or the other before the study. Majority (62.5%) of those who had been tested went for the screening just to know their HIV status. Premarital testing (18.8%) was the second commonest reason for taken an HIV test. Majority of the respondents (74.2%) were willing to go for VCT Among those who were not willing to go for VCT the commonest reason given was that they were certain they were not infected. CONCLUSION: This study highlights the need to step up efforts to increase the students' awareness of VCT, deepen their knowledge and create the right attitude towards VCT through the mass media and religious bodies. Teaching on HIV/AIDS and VCT should also be incorporated into the school curriculum. A shift from the present clinic based approach to a more routine and widespread public health model will increase access to VCT Stand alone VCT centres should therefore be cited in educational institutions, community centres, marketplaces, youth friendly centres etc. for proximity to the people. PMID- 17111753 TI - Knowledge and practice of universal precaution in a tertiary health facility. AB - BACKGROUND: The increased risk of health workers to contract HIV, hepatitis B and C viruses in their work place led to the development of universal precaution or infection control policy. This policy where applied, has been found to reduce the risk of contracting these infections in the work place. The aim of this study is to determine the knowledge and practice of this policy in University of Nigeria Teaching Hospital (UNTH), Enugu. METHODS: This study was cross-sectional in design. Subjects were health workers likely to be exposed to body fluid. The study tool was a self administered pre-tested questionnaire. Statistical analysis was done using SPSS version 11.5 software. RESULTS: Two hundred and forty six health workers participated in the study, consisting of 150 females and 96 males. The male to female ratio was 1: 1.6. Majority of the respondents were between ages 20-49 years. One hundred and twenty four (50.4%) of the respondents were aware of universal precaution, while 88 (35.8%) knew the correct definition of universal precaution. Thirty four (13.8%) had received training on universal precaution however no ward attendant was trained. On multiple regression (P = 0.049) and training (P = 0.006) were the variables that were predictive of correct definition of universal precaution. Hands gloves were used by 86.6% of the respondents, 32.9% did not re-sheath needles and 43.9% practiced appropriate hand washing. CONCLUSION: The level of knowledge and compliance with precaution by health workers in UNTH Enugu is low. Low level of training and unequal training exposure among the various occupations contributes to this. PMID- 17111754 TI - Its perspective of adolescent reproductive health behaviour in a suburban town in Nigeria. AB - BACKGROUND: Parent/Adult Child communication has been shown to influence adolescent reproductive health behaviour Adults need to pass correct information across to youths to help them make proper decisions that will promote their health. The objective of this study was to assess the attitude of parents to adolescent reproductive health behaviour in a suburban Nigerian town. METHODS: This cross-sectional descriptive study was carried out in Ikenne, Ogun State. A Semi-structured questionnaire was used to gather information from 370 adults who were chosen by multi-staged sampling method. RESULT: About two-thirds of the adults surveyed (62.2%) were against premarital sex among adolescents. Although 46.8% said it was permissible for adolescents to use contraceptives, 87.1% will react negatively if they found contraceptives with their children. The condom was the most commonly approved contraceptive method for adolescents by 60.7% of those who approved of adolescents use of contraception, consisting of 14.6% of all the parents in the study. The major reasons against contraceptive use by adolescent were the promotion of promiscuity (41.5%) and infertility in the future (24.9%). Although gender difference was not statistically significant, the females expressed a more conservative attitude to adolescent sexuality and contraception than males; they were less likely to approve of sexual activity among young people (17.8% versus 23.6%) or contraceptive use by them (44.6% versus 49.2%). Eighty seven percent (87%) of those that had children/ward above 12 years had discussed sexuality issues with them. The major form of advice was sexual abstinence and the possibility of unplanned pregnancy. Only 8.5% had advised them about contraception. While 74.1% had approved of sex education in schools, only 31.9% approved of teaching about condom in schools. CONCLUSION: Adults are interested in the reproductive health of their children. Programmes that seek to involve parents in promoting the sexual health of youths will make significant progress if they specifically target parents (especially women) with information about the context of adolescent sexuality and address their misgivings. PMID- 17111755 TI - Men's knowledge of and attitude with respect to family planning in a suburban Nigerian community. AB - BACKGROUND: Men's Knowledge of and attitudes to family planning (FP) in suburban and rural Nigeria is still poor despite a global move to increase the involvement of men in reproductive health matters. A cross-sectional survey was conducted to determine men's knowledge of and attitude to family planning at Ganmo, a sub urban community on the outskirts of Ilorin, Nigeria. METHOD: The study employed an interviewer administered semi-structured questionnaire to elicit information from 360 men in the households. Only males above the age of 15 years resident in the community were selected for interview A proportionate sampling procedure was employed in selecting the required numberof men from each of the 32 compounds that make up the community. RESULTS: Nearly all men (96.5%) were aware of family planning and a majority of them were aware of some common methods of family planning e.g. Oral Contraceptive Pills (OCPs) (72.5%), Injectables (69.2%), Condoms (86.6%) and Traditional methods (70.6%). Knowledge of other alternative female methods was low e.g. Norplant (17.5%), IUCD (26.3%), Diaphragm (39.8%), Vaginal cream (30.2%), Vaginal tablet (37.8%) and Vaginal sponge (16.8%), and Tubal Ligation (51.3%). Knowledge of male controlled FP methods like Withdrawal (49.6%), Rhythm or periodic abstinence (54.6%) and Vasectomy (28.6%) was also poor. The Respondents had low knowledge of common side effects of FP methods e.g. nausea (9.8%), vomiting (13.1%), abnormal menstruation (34.4%), pain (23.2%) and unwanted weight gain (17.0%); some 25.3%% of respondents had no knowledge of any side effects. The attitude of respondents to family planning was also relatively poor as only a moderate proportion of men supported the FP concept (52.7%) and the Nigerian Population Policy (54.8%) of "four children to a woman". Some 54.8% of respondents were in support of men discussing about FP with their spouses. The major reasons for non-approval of FP by men were the fear of side-effects (70.4%) and perception of FP as being against religion (52.1%). The predictors of poor FP attitude were not having formal education, practice of polygyny and to a lesser extent being a Muslim. CONCLUSION: The study concluded that, men at Ganmo have limited knowledge of, and poor attitude to FP An intensive drive at a community based adult reproductive health education was advocated among other recommendations. PMID- 17111756 TI - Health managers' perception of the primary health care management information system: a case of Bama Local Government in northern Nigeria. AB - BACKGROUND: Evaluating the quality and performance of Primary Health Care (PHC) systems depend on the information system's capacity to generate reliable and accurate information, within social, cultural, and economic context. This paper reports an assessment of a PHC health management information system from PHC Managers'perspectives, METHODS: An adapted 3-part Donabedian model informed our assessment of the structure, process and outcomes of the PHC health information system. Pre-tested, semi-structured questionnaires were administered to the PHC Coordinator, 6 Deputy Coordinators, and 18 officers responsible for the health facilities in Bama Local Government Area of Borno State. RESULTS: Majority of the respondents (n=11) believed that staffing at PHC level was inadequate. Only 5 (27.8%) of the managers had training specific to completing HMIS forms. All the facilities were reported to possess registers for the study year (1993), but their numbers dropped by half consecutively down the preceding years to 1990. None reported a health facility that had a copy of the requisite M&E manual guide to HIMIS. Nonetheless 14 reported that report submissions were timely; chief factors causing delays were lack of transport (35.5%), bad roads (16.1%), and scarcity of forms (9.7%). Twelve (12) of the managers judged that the data collected were always or sometimes accurate. Though only 5 crosschecked data to verify accuracy of the submissions. Eight (8) were of the opinion that computerisation was not necessary for rural PHC information system, and eleven (11) felt that the Bama PHC was not ready for computerisation. Twelve (12) of them felt that the quality of the PHC information system had improved since its devolution to the LGA, however, the main suggestions offered to improve the MIS in general were personnel training (32%), feedback from higher levels (20%), and availability of transportation (16%). CONCLUSION: The information system is only as good as the organisation it serves. Results of this study show majorgaps in the structure of the HMIS at the PHC level which is responsible for gathering data onward to the federal level that culminates in epidemiological and health information for the country. Emphasis for intervention for strengthening information systems should be on starting with generating information for local use, and building local capacity to utilise derived information for daily PHC planning, decision-making and management before the prospect of collecting data for upward submission to higher levels. PMID- 17111757 TI - Determinants of neonatal mortality at Wesley Guild Hospital, Ilesa, Nigeria. AB - BACKGROUND: More than 10 million under five children die each year of preventable and easily treatable conditions in developing countries. Of these, eight million are infants, half of whom are newborns in their first month of life. A high proportion of babies die in their first month of life, many of them during their first week. The objective of this study is to assess sociodemographic and other determinants of neonatal mortality in Wesley Guild Hospital (WGH), Ilesa, Nigeria. METHODS: This is a record review of 235 neonatal deaths reported at WGH from January 01 2001 to December 31 2003. Similarly, records of equal number of neonates (235) admitted to the same hospital during the same period but who were discharged alive was also reviewed for comparison. Four hundred and seventy records were reviewed. The two groups were matched for age, sex and within a 7 day period of admission. Information was collected with the aid of predesigned schedule from the patients' case notes, death registers and discharge summaries in the Records Department of the hospital. Information collected included the bio data of the mothers, birth weight of neonates, estimated gestational age at delivery, age at death or discharge, date of admission, duration of the illness and date of discharge. Others included mode and place of delivery, maternal booking status and complications of pregnancy and birth. Data were analyzed using descriptive and inferential statistics by computer software, Epi-Info 2002. RESULTS: Teenage pregnancy, low birth weights (LBW), prematurity and neonatal tetanus were positively associated with neonatal death. Unbooked mothers, deliveries at missions and homes and low socioeconomic status were also positively associated with neonatal death (P < 0.05 in all cases). There was no statistically significant association between the sex of neonate, parity of mother and complications in pregnancy with neonatal death (P > 0.05 in all cases). CONCLUSION: The major determinants of neonatal deaths were teenage pregnancy, prematurity, LBW, poverty and lack of skilled attendance at delivery. Addressing the basic determinants of neonatal mortality will improve newborn survival and health and this will significantly reduce mortality among under five children in developing countries. PMID- 17111758 TI - The use of antimalaria drugs and insecticide treated nets in Ile-Ife, Nigeria. AB - BACKGROUND: The Roll Back Malaria Initiative (RBMI) is aimed at halving the malaria mortality in the year 2010 using prompt diagnosis and treatment of malaria and the use of Insecticide Treated Nets (ITNs). METHODS: This is a cross sectional study of children aged 1 month to 14 years with confirmed malaria to assess the burden of malaria and to evaluate the home use of anti-malarial drugs and ITNs among children with malaria in Ile Ife, Nigeria. RESULTS: Of the total of 1047 children seen, 190 (18.1%) had malaria and were surveyed. Complications were present among 74 (38.9%) children and a significantly higher of the under fives had complications (p by Fisher's exact test = 0.0018). Chloroquine was administered to 63 (67.0%) children within the first 24 hours of the illness although only 25 (26.6%) children had the correct dosage. One hundred and fifty three (80.5%) were aware of the use of untreated bed nets in preventing malaria while 38 (20.0%) and 4 (2.1%) were aware of ITNs and used ITNs respectively. The awareness of ITNs increased with increasing maternal education. CONCLUSION: Awareness of the parents about ITNs and their use are poor. Health education on appropriate antimalarial drug use and ITN use should be intensified. ITNs should be made more widely available. PMID- 17111759 TI - An evaluation of the informed consent process for elective surgery at a university hospital. AB - BACKGROUND: To evaluate on a limited scale the process and quality of the consenting process in our local environment following the increasingly important issue of informed consent and its validity to most physicians and hospital administrators in the face of an emerging litigious Nigeria society. METHODS: A 22 item questionnaire was administered on 60 consecutive patients admitted for elective surgery into the general and specialty surgical wards of the University College Hospital over a one month period. RESULTS: No question in our study achieved the ideal standard of 100%. At the time of admission, 57 of the 60 patients knew their clinical diagnosis from the primary surgical team with 90% (n=54) of the subjects expressing varying degrees of satisfaction with the information they had on their condition. Only about 32% of the patients got additional information on the diagnosis and planned surgery from sources other than the admitting surgical team. In this group of participants there was no significant difference in the sufficiency of information obtained between the two sources (p > 0.05) Ninety percent of the subjects however preferred they had all the information about their operation much earlier than the pre-operation day. Patients satisfaction with overall information obtained during the consenting process was only 'very sufficient' in 35% of the cases. CONCLUSIONS: Notwithstanding the information derived by patients from their primary surgeons in addition to alternate sources, satisfaction with overall information obtained was marginal at 35%. There is a need to look into methods of improving the process, quality and validity of informed consent. PMID- 17111760 TI - Ophthalmic theatre time utilization in a Nigerian teaching hospital. AB - BACKGROUND: An audit of the utilization of the ophthalmic theatre time of the University of Nigeria Teaching Hospital, Enugu was conducted in order to identify the main factors hindering maximum utilization of the theatre, with a view to improving efficiency and effectiveness of the operating theatre team. METHODS: Data was collected prospectively over a period of 3 months (16th May - 22nd August 2005). A total of 28 elective operation lists consisting of 47 eye surgeries was surveyed. Theatre time utilization was studied with respect to commencement and end of cases, turnover interval, start delay interval and main reasons for delays. RESULTS: A total of 57 hrs 5 mins (46.8% of total theatre time used) was spent on doing the actual surgeries and the turnover activities between cases. The total amount of time lost before the commencement of the lists was 64 hrs 50 mins (53.2% of total theatre time surveyed). Late arrival of operating theatre team personnel was noted to be the main single contributory cause of start delays, accounting for 32 hrs 35 mins of lost time. CONCLUSION: The ophthalmic theatre time at our centre is grossly underutilized and measures aimed at reducing start delays and the logistic problems would help ensure optimal utilization of time and other resources. PMID- 17111761 TI - Ophthalmic survey of an old people's home in Nigeria. AB - BACKGROUND: Old people's home is a relatively new experience in Nigeria. Data on the pattern of ocular morbidity among inmates are examined with the aim of making suitable recommendations. METHOD: An old people's home in Port Harcourt was the site of this survey carried out in February 2004. The following data was collected from all inmates-name, sex, age, length of stay, visual acuity, main cause of ocular morbidity, past medical history. Intraocular pressure was measured as indicated. Refractions were carried out on the spot. RESULTS: The male to female ratio of the 20 inmates was 2:3. The age range was from 70-105 years with a mean of 85 years, 7 months +/- 12.14 (SD). The mean duration of stay was 7 years with 50% staying there less than 5 years. 60% had associated systemic illnesses. 85% had visual acuity of less than 3/60 in the better eye. Cataract was the cause of 40% of the ocular morbidity and also caused 57% of the blindness. CONCLUSION: This study emphasizes the need for regular check up of our geriatric population to enable early detection of ocular health problems and thus prevent avoidable disability and dependency. Social support and health insurance could also improve outcome and uptake of treatment options. PMID- 17111762 TI - Congenital eye diseases at Olabisi Onabanjo University Teaching Hospital, Sagamu, Nigeria. AB - BACKGROUND: Congenital eye disorders, though rare are important causes of childhood blindness. It can occur in isolation or in combination, or as part of a syndrome. This retrospective study was aimed at documenting the causes of congenital eye diseases at Olabisi Onabanjo University Teaching Hospital (OOUTH), Sagamu, Nigeria. METHOD: A review of the case notes of patients presenting at the eye clinic with congenital eye diseases between January 1994 and December 2004 was carried out. RESULT: The most common congenital disorders are cataract 50 (47.6%), congenital glaucoma 15 (14.3%), Dacryostenosis 11 (10.5%), and corneal opacity 6 (5.7%) which are causes of preventable blindness. Less common congenital disorders are microcornea (1%), aniridia (1%), retinal atrophy (1%), and congenital anomaly of the optic disc (1%), which are congenital causes of irreversible childhood blindness. CONCLUSION: We conclude that screening programmes should be instituted at the maternity centers before babies are discharged for early detection of congenital eye diseases and treatment of those that can cause preventable blindness. Also we recommend that Government should strengthen our welfare system by providing adequate measures for rehabilitation and care of those with irreversible blindness. PMID- 17111763 TI - Groin flap for coverage of upper extremity defects in Enugu, Nigeria. AB - BACKGROUND: Extensive tissue loss of the hand has remained a big challenge to the Plastic and Reconstructive Surgeon. The objective of this study is to review the use of pedicle groin flap for coverage of distal forearm and hand defects in our centre, and to find out problems of using the flap and make suggestions for improvement. METHOD: This is a retrospective study done using case records of patients treated with pedicle groin flaps for upper extremity defects at the National Orthopaedic Hospital Enugu from January 1999 to December 2004 (six year period). Information on biodata, clinical presentation, treatment, complications and outcome was extracted and analyzed using descriptive statistics. RESULTS: Thirteen (13) patient's case files met the inclusion criteria and were analyzed. There were seven (7) males and six (6) females with mean age ranges between 9 years and 42 years. Four (4) had distal forearm and nine (9) hand wound coverage's. Flap survival was very good to excellent in all cases and the outcome quite satisfactory. CONCLUSION: Use of pedicle groin flap for coverage of defects of the hand and distal forearm remains a very valuable treatment option in our environment. PMID- 17111764 TI - Epistaxis in Port Harcourt. AB - BACKGROUND: Epistaxis often causes significant anxiety in both patients and clinicians. Various methods of treatment have been advocated. The aim of this paper is to document the pattern of epistaxis in Port Harcourt, determine the etiologies and advocate the best treatment option. METHODS: This is a 6-year retrospective study of patients that were managed for epistaxis in the department of Otorhinolaryngology of the University of Port Harcourt Teaching Hospital, Port Harcourt, from January 2000 to December, 2005. The material resources were the records of the Otorhinolaryngology outpatient clinic and Accident & Emergency department of the hospital. These were updated with records from the ward admissions and theatre. The diagnosis of epistaxis was based on clinical history, physical findings, laboratory and radiological investigations with examination under anaestheasia of the nose, nasopharynx and biopsy. RESULTS: There were 30 patients, 16 males and 14 females with a mean age of 30.48 years +/- 15.14 [range 1-70 years]. The most commonly affected age group was 31-40 years. Idiopathic causes accounted for 66.67%, trauma 13.33%, nasopharyngeal carcinoma 10%, chronic maxillary sinusitis 6.67%, and vicarious menstruation 3.33%. The right nasal cavity was more affected than the left. The most commonly used method of treatment was anterior nasal packing (50%). Posterior nasal packing accounted for 26.67% while chemical cauterization was done in 13.3% of cases. CONCLUSION: The pattern of epistaxis in our study agrees with the pattern found elsewhere. Anterior and posterior nasal packing were cost effective methods of treatment. PMID- 17111765 TI - Foreign bodies in the nose. AB - BACKGROUND: Nasal foreign bodies occur most frequently in children and most times first present to the emergency physicians. It poses a challenge to the Otorhinolaryngologist when tampered with. The objective of this paper is to determine the pattern of foreign bodies lodged in the nasal passages in Port Harcourt with a view to documenting the types and highlighting the results of management. METHODS: A retrospective study was done using 134 patients who presented with foreign bodies in the nose to the Ear, Nose and Throat clinic of the University of Port Harcourt Teaching Hospital (UPTH) and Benok Consultants Clinic both in Port Harcourt over an eight-year period from January 1993 to December 2000. RESULTS: One hundred and thirty four patients were seen with a male female ratio of approximately 1:1. Age range was 1-21 years with a mean of 3.23 years +/- 2.48 [SD] and mode of 2-years. The children within the age group 2 4 years had the highest incidence. The right side of the nose was more involved than the left with only one bilateral case noted. The foam was the commonest foreign body found. Most patients presented within the 1st day with only 15 presenting late. Most were removed in the clinic under good illumination, only 6 had to undergo examination under anaesthesia (EUA) in the theatre and these were the uncooperative patients. CONCLUSION: Foreign body in the nose is an emergency only as far as it concerns anxiety to the relations. It presents no problems in management to the Otorhinolaryngologist PMID- 17111766 TI - Plain radiological profile of paranasal sinuses in chronic nasal diseases in University of Port Harcourt Teaching Hospital. AB - BACKGROUND: The paranasal sinuses can get affected by chronic diseases that maybe mitotic, bacterial, fungal and allergic. Plain x-rays, sinoscopy, tomograms, ultrasound scan, CT-scan and magnetic resonance imaging (MRI) technique are some of the means by which the sinus can be evaluated and a diagnosis made. The aim of this study is to evaluate the plain radiological findings of the paranasal Sinuses in-patient with chronic nasal diseases in the University of Port Harcourt Teaching hospital. METHODS: This is a prospective study of patients seen at the otorhinolaryngology out patient clinic of the university of Port Harcourt Teaching Hospital from August 2002 to November 2003. All patients with symptoms suggestive of nasal or paranasal sinus disease of more than 8 weeks duration were evaluated for the study. All the patients had plain radiographs in the occipito mental, occipito-frontal and lateral views. A radiologist interpreted the x-rays. RESULTS: A total of 66 patients were seen with a male to female ratio of 1: 1.4. Ages of patients ranged from 17 years to 65 years. 72.7% had chronic infective sinusitis, 9.1% had vasomotor rhinitis and 6.1% had allergic rhinitis. Plain x rays showed engorged turbinates due to allergy in 43.9% of patients, chronic sinusitis with engorged turbinates in 15.1%, engorged turbinates in 13.6%, chronic sinusitis in 10.6% and opaque nasal fossa in 6.1%. The maxillary sinus was most commonly involved site of disease as seen in (71.2%) of patients. The sphenoid sinus was the least involved (7.4%). Thirteen patients (19.7%) did not show any changes in the sinus x-rays. CONCLUSION: Chronic sinusitis is the commonest chronic nasal disease. Engorged nasal turbinate was the commonest plain x-ray finding. The maxillary sinus was the most commonly diseased sinus. In 19.7%, the plain radiograph was normal. Plain radiographs are a cheap and readily available tool for the investigation of diseases of the paranasal sinuses with good results in this environment. PMID- 17111767 TI - Pathology as a profession: contribution of undergraduate training on choice of career among final year students of a medical school in north central Nigeria. AB - BACKGROUND: The choice of pathology as a profession by would be resident doctors, will, to a large extent depend on the amount of knowledge acquired on the subject at the undergraduate level. METHODS: A questionnaire was self administered to final year medical students of University of Jos shortly after the completion of the final written paper in medicine in September 2005; meant to elicit their views on choosing patology as a profession after graduation. RESULTS: Eighty-five students enrolled comprising 65 (76.5%) males and 20 (23.5%) females Ninety six percent of the students were between 26 and 30 years. Those who found Pathology interesting were 76.5%; 18.8% picked pathology as their career of first choice after graduation 5.9%, 3.5% and 5.9% as 2nd, 3rd and 10th choices repectively. Among the pathology disciplines, 8.2%, 4.7%, 3.5% and 2.4% would specialize in Histopathology, Haematology, Chemical Pathology and Medical Microbiology respectively. The students generally complained of inadequate time and exposure during the postings as well as the difficulty in understanding Medical Microbiology. CONCLUSION: More time should be allocated to the teaching of Pathology at the undergraduate level. Learning enhancing aids should be introduced such as: regular practical demonstrations, audio-visual teaching aids, computer assisted programs and the establishment of pathology museums in the medical schools across the country. PMID- 17111768 TI - Comparison of red cell cholinesterase and plasma cholinesterase activities in early detection of organo-phosphorus toxicity in exposed industrial workers in Port Harcourt, Nigeria. AB - BACKGROUND: The Niger Delta region of Nigeria has a high concentration of companies using organo-phosphorus compounds with workers who have been exposed to these compounds for varying lengths of time resulting in toxic manifestations in some of the workers. Studies have revealed that plasma cholinesterase and red cell cholinesterase levels are reduced following organo-phosphorus toxicity. This study looks at cholinesterase levels in plasma and red cell in the occupationally exposed groups and control with a view to determining which will be more useful in the early detection of organo-phosphorus toxicity. METHOD: Fourty seven subjects with early symptoms of organo-phosphorus toxicity who have worked for more than 4 years with organo-phosphorus compounds were selected from five companies localized around the same area in Port Harcourt. Fifty subjects who do not work directly with organo-phosporus compounds from the same companies as the 47 workers above but not working directly in the organo-phosphorus units were also studied while 50 persons working in the civil service were recruited as controls. The kinetic colorimetric method was used for measuring plasma and red cell cholinesterase activity Result: The results showed significant reduction in the mean plasma cholinesterase activity in the chronically exposed subjects (4614. +/- 532) who showed symptoms of toxicity when compared with that of the control group (8095 +/- 575). There was no statistical difference in the mean red cell cholinesterase activity of the chronically exposed group (7998. +/- 948) when compared with that of the control (8115. +/- 712) P > 0.05 CONCLUSION: The effect of organo-phosphorus poisoning occurs early and is more marked in plasma cholinesterase than in the red cell cholinesterase. Plasma cholinesterase is thus more useful in early detection of organo-phosphorus toxicity than red cell cholinesterase. PMID- 17111769 TI - Biochemical and haematological assessment of workers exposed to some petroleum products in Rnugu Urban, Rnugu State, Nigeria. AB - BACKGROUND: Exposure to petroleum product(s) occurs during extraction, refining, transportation and utilisation. Surprisingly, this relatively common hazard has received little public recognition. We therefore studied the adverse effects of petroleum products among occupationally exposed liquefied petroleum gas and histochemical workers in Enugu Urban, Nigeria. METHODS: Eight industries/departments were surveyed. Sixty workers were tested for some biochemical and haematological parameters (phosphatases, transaminases, complete blood count, erythrocyte sedimentation rate), were analysed, using standard methods. Also, 30 age and sex-matched apparently healthy subjects served as the control group. RESULTS: When the test and control results were compared, there were statistically significant differences (p < 0.05) in: (1) haematocrit, reticulocytes, erythrocyte sedimentation rate, platelet, lymphocyte, monocyte, alkaline phosphatase, aspartate transaminase, alanine transaminase among liquefied petroleum gas workers and (2) haemoglobin, haematocrit, erythrocyte sedimentation rate, platelets, neutrophil, lymphocyte, monocytes, acid phosphatases and alkaline phosphatase among histochemical workers. Overall, there was statistically significant decrease (p < 0.05) in most of the parameters among the test subjects when compared with the control group. Blood films of the workers revealed mild to moderate hypochromia, anisocytosis and poikilocytosis while the controls were normocytic and normochromic. CONCLUSION: We conclude that exposure to petroleum products adversely affects biochemical and haematological parameters. Hence, improved working conditions are recommended. PMID- 17111770 TI - The endocannabinoid system: association with metabolic disorders and tobacco dependence. AB - RACKGROUND: Obesity and smoking are major health problems globally as they are risk factors for cardiovascular disease. There appears to be a link between obesity and smoking which is manifested in the endocannabiniod system. This review aims to highlight the physiologic role of the endocannabinoid system and its possible therapeutic applications. METHODS: Literature on the endocannabiniod system was reviewed using MEDLINE/PUBMED and bibliographic searches for English language studies. RESULT: The endocannabiniod system is a physiological system that plays a significant role in the regulation of body metabolism, especially glucose and fat metabolism in addition to nicotine dependence with promising therapeutic application. CONCLUSION: The discovery and insights into the functioning of the endocannabiniod system have made a way for new and effective therapeutic approaches. These therapies may provide an effective treatment for obesity and related risk factors for diabetes and cardiovascular disease as well as help people to stop smoking. PMID- 17111771 TI - Culture and health: the effect of Nupe cultural practice on the health of Nupe people. AB - BACKGROUND: The Cultural practices of communities are known to influence the Health status of the community both positively and negatively. To achieve set out health goals positive cultural practices should be enhanced and incorporated into community based health programmes. This commentary aims to highlight the positive and negative effects of cultural practices on health using the Nupes a tribe in North central Nigeria as a reference point. METHOD: Information on the cultural practice of Nupe people and the related health effects were obtained through observation, group discussion and interviews among Katcha people, a Nupe community in Nigerstate of Nigeria. Literature of the effects of cultural practices on health was reviewed using MEDLINE and manual library search. RESULTS: Cultural practices with positive health effects such a "Tamako" a system of community based assistance to the sick was found among the Nupes in Katcha. This system is useful in defraying hospital bills of indigent members of the community. Another positive practice is the culture of food assistance and gifts to nursing mothers which helps to improve their nutritional status. However Negative cultural practices such as child marriage, "Sadakiar" (wife gifts), "Egikpa" (child fostering) and "Efidan" (body scarifications) are also practiced. CONCLUSION: Cultural practices have significant effects on health. Most of these effects are detrimental and should be discouraged. Cultural practices with positive effects should be encouraged and integrated into community based health policies and programmes in order to enhance the attainment of the millennium development goals especially in rural communities of the developing world. PMID- 17111772 TI - Facioscapulohumeral muscular dystrophy (landouzy-dejerine type) in a Nigerian female: a case report. AB - BACKGROUND: Muscular dystrophy is not an uncommon entity in Nigerian medical clinics. The facioscapulohumeral type represents a rare variety of the disorder with its own distinctive characteristics but is not expected to have significant cardiac manifestations. METHOD: The case report of a 17-year-old Nigerian female with facioscapulohumeral muscular dystrophy and significant cardiac dysfunction is presented and the relevant literature is reviewed. RESULTS: A 17-year-old Nigerian female presented with 18 month history of shortness of breath on mild to moderate exertion, generalized weakness, weight loss and abnormal gait. Leg swelling developed a few weeks prior to presentation. Intrauterine life and early childhood were uneventful while her family history was unremarkable. She had typical features of facioscapulohumeral muscular dystrophy but in addition demonstrated evidence of significant cardiac impairment, which is uncommon and not typically expected in this disorder. CONCLUSION: It is important for clinicians to comprehensively evaluate every patient presenting with a hitherto "clear "diagnosis in order to unmask unexpected associated clinical details. PMID- 17111773 TI - Parkinsonism induced by sepsis: a case report. AB - BACKGROUND: Parkinson's disease (PD) is a slowly progressive neurodegenerative disease that appears essentially as a sporadic condition with no identifiable cause. Parkinsonism is used for syndromes where the aetiolobgy is known such as Parkinsonism due to stroke, infection, neuroleptic drugs and toxic agents. Parkinson's disease and Parkinsonism present with the tetrad of tremor at rest, slowness of voluntary movement (bradykinesia), rigidity and a characteristic disturbance of gait and posture. A report of Parkinsonism induced by sepsis is rare. This report aims to create awareness of Parkinsonism as a manifestation of sepsis. METHOD: The case note of a patient with Parkinsonism induced by sepsis managed in the medical unit of the University of Port Harcourt Teaching Hospital and a review of the literature on the subject with Medline search was used. RESULT: A 71-year-old Nigerian male presented with Parkinsonism on a background of Gram negative sepsis which resolved with antibiotic therapy. Antiparkinoinian drugs were not used. CONCLUSION: Parkinsonism is a rarely reported neurological complication of sepsis. There is a need for physicians to be aware of this clinical manifestation. PMID- 17111774 TI - Osteonecrosis complicating chronic myeloid leukaemia. AB - BACKGROUND: There are previous reports of osteonecrosis among Nigerians with sickle cell diseases and some other medical conditions. To the best of the authors' knowledge there is, however, no known such report attributed to chronic myeloid leukaemia, especially in Nigeria. Apart from presenting this as the first report, it is also to emphasize the need for early full blood count in persons presenting with otherwise non-haematological condition. METHOD: The case note of the patient and the available literature on the subject were used. RESULT: The 15 year-old girl who presented with a fifteen-month history of right hip joint pain and progressive abdominal swelling with weight loss for five months was found to have chronic myeloid leukaemia and avascular necrosis of the right femoral head. Her gait and walk improved remarkably within two weeks of commencing cytoreductive therapy. However, she still uses crutches due to the extensive bone damage from the prolonged impaired blood supply caused by chronic myeloid leukaemia before presentation. CONCLUSION: Osteonecrosis of the femoral head can complicate chronic myeloid leukaemia from hyperviscosity-induced leucostasis. Early simple peripheral blood examination in diseases presenting with otherwise non-haematological symptoms will reduce, if not prevent, severe complications of diseases. PMID- 17111775 TI - Ileal carcinoid. AB - BACKGROUND: Gastrointestinal carcinoids are ill understood, enigmatic malignancies which are extremely difficult to diagnose. In spite of their slow growth rate, they can be aggressive. The aim of this report is to present a case of ileal carcinoid which produced a complete intestinal obstruction. METHOD: The case records of a 78-year-old tailor who was treated over a period of one year and diagnosed with an ileal carcinoid and a review of literature using medline and manual library search were used. RESULT: The patient was admitted and treated conservatively on two occasions in 2004 with features of subacute intestinal obstruction. He improved on each occasion and was discharged. He was admitted for the third time in 2005 with complete intestinal obstruction which required an exploratory laparotomy. A small, hard and completely obstructing tumor was found in midileum. No synchronous tumor was detected and there was no ascites. The liver felt and looked normal. Ileal resection and end-to-end anestomosis was done. Post operative recovery was complicated by a bout of severe diarrhea which required intravenous fluid therapy. There was good wound healing and was discharged 12 days after laparotomy. He was well clinically at follow-up review one month afterdischarge. CONCLUSION: The case illustrates the difficulty in making a diagnosis of this condition before surgery. PMID- 17111776 TI - Suspected epidermolysis bullosa simplex in a 5-week-old Nigerian: a case report. AB - BACKGROUND: Epidermolysis bullosa is a rare non scaring autosomal dominant disorder characterized by recurrent blistering of the skin and mucous membrane. The skin is fragile and minor rubbing may cause blistering. It's epidemiology in our environment is unknown probably because of paucity of information on the clinical presentation and management with resultant mortality within the first few months of life. METHOD: A case report of a 5-week-old female who presented with generalized blistering and denudation of the skin first noticed on the left foot at birth and a review of the literature on the subject using Medline and online search was used. She was treated at various traditional medicine homes and clinics before referral to the teaching hospital. She was managed initially for bullous pemphigus with antibiotics for proven septicaemia, and the wound infection ignorantly managed with daily sofratulle dressing alternating with closed dressings before a definitive clinical diagnosis of was made. RESULT: The patient was referred late to the tertiary centre. She was initially treated for bullous pemphigus and sepsis with antibiotics and wound dressings with poor response before the diagnosis of epidermlysis bullosa was made. The patient died from severe foot bleeding before blood could be transfused. CONCLUSION: Delay in making the right diagnosis hence the appropriate treatment even in a tertiary health is being highlighted. PMID- 17111777 TI - The need for high index of suspicion in early diagnosis of typhoid fever in young children. AB - BACKGROUND: The diagnosis of typhoid fever in young children is more difficult than in the adult due to the unusual pattern of presentation. This letter aims to sensitize general physicians and paediatricians on the need for a high index of suspicion in the diagnosis of typhoid fever in children. METHOD: The case records of a child diagnosed with typhoid fever and a review of literature on the subject were used. RESULT: A five year old female misdiagnosed as a case of severe malaria and sepsis was found to have typhoid fever on post mortem examination. CONCLUSION: It is important to make an early diagnosis of typhoid fever in young children in order to prevent mortality from this treatable disease. PMID- 17111778 TI - [Confused Germanic blasphemy. Jacob Moleschott and materialistic medicine]. AB - Starting from the reading of a recent biography of the Dutch materialistic physiologist Jacopo Moleschott (1822-1893), this article proposes a brief survey of the impact, especially in Italy, of the materialistic paradigm, not only in the field of medicine and of the natural sciences, but also in that of philosophy and of literature. From the rejection and ferocious criticisms of the Jesuits, such as Padre Previti, to the harsh ironies of Tommaseo--who, in reference to the academic lessons of Moleschott, spoke of "confused Germanic blasphemy"--, the survey proceeds to a recognition of the role and influence of Moleschott's perspective in the field of medicine, in that of science, and especially in the philosophical and epistemological debate on the relation between the experimental sciences and philosophy. The survey then proceeds to touch on the classical controversy over the Chemische Briefe of Liebig (1844), the dispute with Bufalini on the new way of thinking about the relation between physiology and pathology, and the influence exerted by the Dutch physiologist on Salvatore Tommasi. The article in the end broadens to include a rapid analysis of Moleschott's contribution to the field of literature, as well as to the more complicated debate on the natural sciences and materialism, which still today presents, beyond the outdated models of positivistic scientism, well-grounded themes of interest, if not of validity. PMID- 17111779 TI - The faster the better? PMID- 17111780 TI - "Bridging" therapy in patients on long-term vitamin K antagonist treatment: a yet unsolved issue. PMID- 17111781 TI - The management of patients who require temporary reversal of vitamin K antagonists for surgery: a practical guide for clinicians. AB - The management of patients who require temporary interruption of vitamin K antagonists is a common clinical problem, affecting an estimated 400 000 patients per year in Europe and North America. Managing such patients is challenging because of the lack of randomized trials assessing different perioperative anticoagulation management strategies and inconsistent recommendations from consensus groups. Recent non-randomized trials have helped to estimate the risks for arterial thromboembolism and bleeding with bridging anticoagulation involving low-molecular-weight heparin. The objectives of this review are to describe bridging anticoagulation and how it may be used with a short-acting heparin, such as unfractionated heparin or low-molecular-weight heparin, to discuss preoperative patient management, focusing on risk stratification for thromboembolic events and interruption of vitamin K antagonist therapy, and to discuss postoperative patient management, focusing on surgery-related bleeding risk and the resumption of bridging anticoagulation and vitamin K antagonist therapy. PMID- 17111782 TI - Takayasu's arteritis: a review of the literature. AB - Takayasu's arteritis is a rare, idiopathic, chronic inflammatory disease with cell-mediated inflammation, involving mainly the aorta and its major branches. It leads to stenosis, occlusion or aneurysmal degeneration of large arteries. The clinical presentation is characterised by an acute phase with constitutional symptoms, followed, months or years later, by a chronic phase in which symptoms relate to fibrosis or occlusion of vessels. Angiography is the gold standard for diagnosis and for topographical classification and it correlates with symptoms and prognosis. Here we focus on the pathophysiology, clinical and angiographical classification, diagnostic assessment and therapeutic approach of Takayasu's arteritis. PMID- 17111783 TI - Daily dose of interferon alpha-2b and ribavirin in treatment-naive patients with chronic hepatitis C virus genotype 1 infection: a randomised controlled study. AB - OBJECTIVE: The treatment of patients with hepatitis C virus (HCV) genotype 1 infection remains disappointing. METHODS: In 1999, we started a multicentre study comparing two regimens of recombinant interferon (IFN) alpha-2b plus ribavirin. Group A (90 patients) received ribavirin plus IFN alpha-2b 5 MU/day for 1 month (induction therapy) followed by IFN alpha-2b 5 MU thrice weekly for 5 months. Group B (85 patients) received ribavirin plus IFN alpha-2b 5 MU thrice weekly for 6 months. Responders in both arms received IFN alpha-2b 3 MU thrice weekly for a further 6 months. A follow-up evaluation was performed at 18 months. RESULTS: One hundred and seventy-five consecutive treatment-naive patients with HCV genotype 1 infection were enrolled in the study. A sustained virological response (SVR) was obtained in 51 (29%) patients: 28 in group A (31%) and 23 in group B (27%). HCV RNA clearance was greater at 3 months among patients who received induction therapy (57 vs 39%; p < 0.02). Age, sex, and initial viral load did not influence the achievement of a SVR. HCV clearance at the end of the study was lower in cirrhotic patients (3/26 vs 48/149; p < 0.05). The only SVR in patients with cirrhosis occurred in those from group A (p < 0.05). Both regimens were well tolerated. CONCLUSIONS: This study confirms the low rate of SVR in treatment naive patients with HCV genotype 1 infection treated with IFN alpha-2b plus ribavirin. A 4-week induction regimen was slightly superior to standard IFN alpha 2b plus ribavirin. Although the number of patients with cirrhosis was low, induction therapy seemed to be more effective in cirrhotics. Given its safety and tolerability, the induction regimen evaluated here may be a therapeutic option in treatment-naive patients with HCV genotype 1 infection. PMID- 17111784 TI - Psychological impact of thrombosis in the young. AB - OBJECTIVE: Thrombosis is rare in the young, but can cause severe psychological distress that influences the quality of life and the coping capacities of these patients. This study was meant to increase the understanding of self-perception, social and family functioning and ways of coping with the disease in young patients after an episode of thrombosis. METHODS: Seven questionnaires spanning social and family functioning and ways of coping with disease were completed by 50 patients < or = 45 years of age after a first episode of venous or arterial thrombosis. Data were compared with those of 39 healthy individuals with similar age, sex and level of education. RESULTS: Compared to healthy controls, young patients with thrombosis had lower self-esteem, showed higher impairment in social activities and in familial relationships, and used more frequently coping strategies. Most patients used all coping strategies, preferring the more active ones, but more patients than controls used the passive ones, particularly "avoidance" and "religiosity". A correlation was found between the frequency of use of passive or negative coping strategies and some of the psychological, social and familial dimensions that scored more negatively. Greater psychological impairment and differences in coping styles were found in women compared with men and in individuals < 34 years compared with those > or = 34 years. CONCLUSIONS: Young individuals with thrombosis develop psychological changes that influence their behaviour, quality of life and coping. This is particularly evident in women and in young patients. Physicians dealing with thrombosis should be aware of this situation and offer psychological support. PMID- 17111785 TI - An unusual case of pulmonary hypertension in a young male. PMID- 17111786 TI - Intermittent claudication or neurogenic claudication? "Why don't you speak to me"? PMID- 17111787 TI - Testing in syncope. PMID- 17111788 TI - Airway management: the sine qua non of emergency medicine. PMID- 17111789 TI - Difficult airway management. AB - Airway management is unequivocally the most important responsibility of the emergency physician. No matter how prepared for the task, no matter what technologies are utilized, there will be cases that are difficult. The most important part of success in the management of a difficult airway is preparation. When the patient is encountered, it is too late to check whether appropriate equipment is available, whether a rescue plan has been in place, and what alternative strategies are available for an immediate response. The following article will review the principles of airway management with an emphasis upon preparation, strategies for preventing or avoiding difficulties, and recommended technical details that hopefully will encourage the reader to be more prepared and technically skillful in practice. PMID- 17111790 TI - The utility of head computed tomography in the emergency department evaluation of syncope. AB - OBJECTIVE: Current guidelines for evaluation of syncope recommend that in the absence of objective focal neurologic findings, head computed tomography (HCT) may not be necessary. Compliance with this recommendation is highly variable, which may be due in part to the lack of currently available evidence. We undertook the following investigation to determine whether HCT aids in the diagnostic investigation of syncope. METHODS: This study was a retrospective chart review of all adult patients who presented to an urban emergency department, and who had a HCT ordered for syncope, during a 6-month period in 2001. Patients with competing indications for HCT, or those with a presentation consistent with seizures were excluded. Charts were assigned to the "positive" or "negative" HCT group depending on whether the treating physician considered HCT findings relevant to the syncopal event. RESULTS: A total of 202 patients had a HCT performed for syncope. Eighty-five patients met one or more of the exclusion criteria. HCT of the remaining 117 patients were analyzed. None of the 117 patients had a HCT finding that was clinically related to the syncopal event. CONCLUSIONS: HCT yielded no relevant clinical findings in our entire sample of patients with syncope. Our findings combined with previous studies add to the growing body of evidence that HCT for syncope in the absence of focal neurologic findings may not be necessary. PMID- 17111791 TI - Airway forum. PMID- 17111792 TI - Headache in a patient with ulcerative colitis. PMID- 17111793 TI - Antibiotic prophylaxis to prevent nosocomial infections in patients in intensive care units: evidence that struggle to convince practising clinicians. AB - BACKGROUND: Pneumonia is an important cause of mortality in intensive care units. The incidence of pneumonia in such patients ranges between 7 and 40%, and the crude mortality from ventilator associated pneumonia may exceed 50%. Although not all deaths in patients with this form of pneumonia are directly attributable to pneumonia, it has been shown to contribute to mortality in intensive care units independently of other factors that are also strongly associated with such deaths. OBJECTIVES: The objective of this review was to assess the effects of antibiotics for preventing respiratory tract infections and overall mortality in adults receiving intensive care. Search strategy. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (issue 3, 2003), which contains the Acute Respiratory Infections (ARI) Group specialised trials register; MEDLINE (January 1966 to September 2003); EMBASE (January 1990 to September 2003); proceedings of scientific meetings and reference lists of articles from January 1984 to December 2002. We also contacted investigators in the field. Selection criteria. Randomised trials of antibiotic prophylaxis for respiratory tract infections and deaths among adult intensive care unit patients. Data collection and analysis. At least two reviewers independently extracted data and assessed trial quality. RESULTS: Overall 36 trials involving 6922 people were included. There was variation in the antibiotics used, patient characteristics and risk of respiratory tract infections and mortality in the control groups. In 17 trials (involving 4295 patients) that tested a combination of topical and systemic antibiotic, the average rates of respiratory tract infections and deaths in the control group were 36% and 29% respectively. There was a significant reduction of both respiratory tract infections (odds ratio 0.35, 95% confidence interval [CI] 0.29-0.41) and total mortality (odds ratio 0.78, 95% CI 0.68-0.89) in the treated group. On average 5 patients needed to be treated to prevent one infection and 21 patients to prevent one death. In 17 trials (involving 2664 patients) that tested topical antimicrobials alone (or comparing topical plus systemic versus systemic alone) the rates of respiratory tract infections and deaths in the control groups were 30 and 26% respectively. There was a significant reduction of respiratory tract infections (odds ratio 0.52, 95% CI 0.43-0.63), but not in total mortality (odds ratio 0.97, 95% CI 0.81-1.16) in the treated group. CONCLUSIONS: A combination of topical and systemic prophylactic antibiotics reduces respiratory tract infections and overall mortality in adult patients receiving intensive care. A treatment based on the use of topical prophylaxis alone reduces respiratory infections, but not mortality. The risk of occurrence of resistance as a negative consequence of antibiotic use was appropriately explored only in the most recent trial by de Jonge, which did not show any such effect. PMID- 17111794 TI - What is the next step in D-dimer research? Education of physicians. PMID- 17111795 TI - Oral anticoagulant therapy and telemedicine. PMID- 17111796 TI - Syncope as a symptom of non-massive pulmonary embolism: a case report. PMID- 17111797 TI - Sarcoid relapse with isolated kidney involvement. PMID- 17111798 TI - "Reversible" left bundle branch block in acute carbon monoxide poisoning. PMID- 17111799 TI - [Diagnosis and treatment and prevention of iatrogenic functional aphonia]. AB - OBJECTIVE: To discuss the diagnosis, treatment and prevention of iatrogenic functional aphonia. METHODS: Twenty three patients who either lost their voice or only could whisper after surgery in other hospitals were included in this study as the first group, history was well collected and laryngostroboscopy performed. All cases were confirmed as iatrogenic functional aphonia patients and received phonation therapy. In another group of patients who received vocal cord surgery in our hospital from 2003 to 2005, speaking was restricted while not prohibited after surgery, voice quality was closely observed, and 1028 cases were included. RESULTS: All 23 cases of functional aphonia were cured with phonation therapy. No iatrogenic functional aphonia occurred in the second group of patients. CONCLUSIONS: The iatrogenic functional aphonia can be caused by post operative mistreatment and could be cured with phonation therapy, and it is preventable if speaking is not strictly prohibited after surgery. PMID- 17111800 TI - [One case report of foreign body in the eustachian tube taken out by extended anterior tympanotomy]. PMID- 17111801 TI - [Clinical characteristics of myasthenia gravis with dysphonia as the initial symptom]. AB - OBJECTIVE: To identify clinical features and diagnostic tests that would alert the otolaryngologist to consider myasthenia gravis (MG) in the differential diagnosis of dysphonia, we reviewed the clinical characteristics of MG whose initial symptom is dysphonia. METHODS: 31 patients who presented with dysphonia as their initial and primary complaint are reported, their symptoms and signs are observed and analyzed. RESULTS: Patients with dysphonia as their initial symptom of MG may complain of vocal fatigue, difficulty sustaining or projecting their voices, breathy voice or intermittent hoarseness. These symptoms are characterized by fluctuating weakness and abnormal fatigability. Flexible fibroendoscopic examination revealed that patients had incomplete adduction of the vocal folds, fatigue of the tensors of the vocal fold, incomplete glottic closure, vocal cord paralysis, saliva pooling over the bilateral or unilateral pyriform sinus. Neostigmine test revealed dramatic improvement in all patients. Serum levels of anti-Ach-R antibodies were tested in 19 cases, only 5 cases were abnormality. All patients had improved after treatment CONCLUSIONS: Voice changes can be the first sign of early MG. Based on fluctuating weakness or weak voice at the end of the day, a positive neostigmine test, significantly higher circulating antibody to acetylcholine receptor, a diagnosis of MG could definitively be made. PMID- 17111802 TI - [Glottic measurement and vocal evaluation after three surgical techniques in the treatment of bilateral vocal cord paralysis]. AB - OBJECTIVE: To evaluate postoperative glottic area and vocal quality of three various surgical techniques for treating bilateral vocal cord paralysis, including laser arytenoidectomy (Group A, 24 cases), reinnervation of the posterior cricoarytenoid muscle by phrenic nerve (Group B, 9 cases) and arytenoidectomy accompanying lateral cordopexy by extralaryngeal approach (Woodman's procedure, Group C, 13 cases). METHODS: 46 cases suffered from bilateral recurrent laryngeal nerve injury were included in our study. The pre postoperative glottic measurement and vocal acoustic parameters were analyzed. RESULTS: The decannulated cases in group A and group B and group C were 22, 8, 13 respectively. The post-operative mean maximal glottic area was (47.2 +/- 7.4) mm2, (78.3 +/- 16.0) mm2, (48.1 +/- 6.5) mm2 respectively. Group B cases glottic area was larger than that of group A and group C (t value were 4.46 and 3.85, P value were 0.000 and 0.001). No significant difference was found between group A and group C (t = 1.68, P = 0.101). After surgery, in group A, 17 cases voice quality was the same compared with that of before surgery, and 7 cases voice quality had become worse; In group B, the voice quality had become better in 5 cases, completely recovered in 1 case, and had not change in 3 cases; In group C, the voice quality had become deteriorated in 10 cases and no change in 3 cases. And in group B, ipsilateral diaphragm paralysis in 9 cases after surgery, whose vital capacity and forced vital capacity had decreased to 72%-84%, 76%-84% of that before the surgery respectively; and the diaphragm mobility had recovered by 35%-76% respectively, while vital capacity and forced vital capacity had become 93%-97%, 91%-98% of that before the surgery. In Group B, all cases' pulmonary function was normal half a year postoperatively. CONCLUSIONS: Reinnervation of the posterior cricoarytenoid muscle by phrenic nerve seems to be best procedure with better post-operative voice and larger glottic area. Although the sufficient airway for decannulation can be acquired in Group A and Group C, but most of patients in Group A had pre-operative vocal level and badly abnormal in Group C. PMID- 17111803 TI - [Laryngeal electromyographic characteristics of vocal fold immobility]. AB - OBJECTIVE: To investigate the characteristics of laryngeal electromyography and evoked electromyography in vocal fold immobility. METHODS: 108 cases of vocal fold immobility were analyzed by clinical manifestation and laryngeal electromyography characteristics, including spontaneous potential activity, motor unit potential (MUP) measurement, recruitment pattern analysis and evoked electromyography evaluation. RESULTS: Neurogenic vocal fold immobility showed a wide variety of abnormal activity. Fibrillation potentials and positive sharp waves were found in patients with laryngeal nerve injuries. For laryngeal paralysis, there was no reaction with LEMG and evoked LEMG. For laryngeal paresis, it showed decreased MUP and decreased recruitment activities with simple pattern recruitment or mixed pattern recruitment, and decreased evoked muscle response potentials were also shown with delayed latency than normal (P < 0.05) in the TA (2.2 +/- 1.0) ms(x +/- s), PCA (2.4 +/- 1.0) ms and lower amplitude in the TA (0.9 +/- 0.7) mV and PCA (1.2 +/- 1.0) mV (P < 0.05). Patients with vocal fold mechanical limitations generally yielded normal LEMG and Evoked LEMG. Patients with neoplastic infiltration of the laryngeal muscles demonstrated decreased LEMG and nearly normal Evoked LEMG with normal latency and lower amplitude. CONCLUSIONS: General EMG and evoked EMG play a crucial role in the diagnosis of vocal fold immobility. PMID- 17111804 TI - [Relationship between laryngeal morphology and voice changes in old people]. AB - OBJECTIVE: To investigate the relationship between laryngeal morphologic changes and voice changes in old people. METHODS: Fifty four healthy people older than 60 years were studied. Laryngeal morphologic changes and vocal acoustics features were observed and analyzed. Forty six college students aged between 21-22 years were included as the young control group, and 23 of them are males and 23 females. RESULTS: Sixteen cases with vocal fold atrophy (55.2%), 18 cases with glottis incompetence (62.1%), 16 cases with hypoglottic mucosal edema (55.2%), 12 cases with surpassing ventricular fold (refers to the phenomenon that the ventricular fold covers part or all of the vocal cord during phonation) (41.4%) were observed in the old male; 15 cases with edema of interarytenoid fold (60.0%), 14 cases with glottis incompetence (56.0%), 12 cases with vocal fold atrophy (48.0%), 8 cases with vocal fold edema (32.0%) were observed in the old female. Fundamental frequency (F 0) ascended in the male while declined in the female; jitter, shimmer and noise to harmonic ratio enlarged, and vocal tremor intensity index increased too. Soft phonation was noticed especially in the voices of the old male. Compared with the control group, there was statistic significance (each p value is less than 0.05 or 0.01) except for the increasing of jitter, shimmer and soft phonation index in the old female. CONCLUSIONS: Significant changes were found in the old when the morphology of larynx and features of vocal acoustics were studied, hypoglottic edema and interarytenoid edema are common in the old. PMID- 17111805 TI - [Epidemiologic study on hearing impairment and ear diseases in old people]. AB - OBJECTIVE: To investigate the prevalence of hearing impairment and ear diseases in old people and provide scientific data for drawing up the prevention and treatment strategies. METHODS: Using the probability proportion to size (PPS) method, 1261 people over 60 years were investigated in 40 clusters in Jiangsu Province with the WHO protocol. RESULTS: The prevalence of hearing impairment was 58.1% (the standardized rate: 59.5% in the whole country, 60.9% in Jiangsu province). Degrees of hearing impairment were mild (33.1%), moderate (17.8%), severe (5.9%) and profound (1.3%). The prevalence of hearing disability was 25.0% (the standardized rate: 26.6% in the whole country, 28.1% in Jiangsu province). There were significant difference of the prevalence between male and female, as well as urban and rural, and different ages. The prevalence of the ear diseases was auricle malformation (0.2%), wax (1.7%), otitis externa (0.1%), fungi (0.5%), serous otitis media (1.2%), chronic suppurative otitis media (1.6%), dry perforation of tympanic membrance (2.3%). The causes of hearing impairment were ear diseases (2.9%), non-infectious condition (92.6%), genetic condition (0.3%) and undetermined causes (4.2%). Of which, 31.1% of persons needed hearing aids while 2.3% of persons needed medicine treatment, but 0.9% of persons needed non urgent surgery and 1.0% of persons needed other treatment. CONCLUSIONS: The prevalence of hearing impairment and disability in the old rised obviously than the last investigation in 1987. It was a heavy burden for social development in China. The government and the whole society should take more concern about the problem. The scientific strategies of prevention and treatment were urgently needed and implemented. PMID- 17111806 TI - [Surgery of jugular foramen region tumors]. AB - OBJECTIVE: To evaluate the various surgical approaches in removing the jugular foramen region tumors in order to obtain the maximal functional preservation. METHODS: Between March 1993 and May 2005, 32 patients with jugular foramen region tumors were operated. The mastoid and neck approach, the infratemporal fossa approach with and without rerouting the facial nerve, the combined transcochlear and infratemporal fossa approach were respectively used according to the tumor type, size, auditory, facial and lower cranial nerve functions. The cranial nerve monitoring was used and a postoperative CT or MRI scanning was taken in all cases. RESULTS: In this series of 32 patients, 13 were paragangliomas, 10 schwannomas,2 meningiomas, 2 giant cell tumors, 1 chondrosarcoma, 1 mucochondrosarcoma, 1 adenoid cystic carcinoma, 1 rhabdomyosarcoma and 1 case cholesterol Granuloma. The follow-up period was 3-60 months. Total removals were achieved in 26 cases, near-total removal in 5 cases, and subtotal removal in one case. One patient died after surgery because of intracranial hemorrhage. Another patient died two year after operation because of recurrence. Postoperative cerebrospinal fluid leakage was occurred in 5 cases and meningitis in two cases. They were all treated conservatively. The facial function one week after operation was grade 1-2 in 13 cases, grade 3-4 in 12 cases and grade 5-6 in 7 cases. It changed into grade 1-2 in 23 cases, grade 3-4 in 7 cases and grade 5-6 in 2 cases. The postoperative hearing was improved in 4 cases, stable in 10 cases, deteriorated in 11 cases and totally lost in 7 cases. The lower cranial nerve function was normal in 9 cases, transient palsy in 11 cases, permanent paralysis with compensation in 10 cases and without compensation in 2 cases. CONCLUSION: The jugular foramen region tumor could be removed with maximal function preservation by using various surgical approaches. The postoperative life quality was acceptable in most cases. Surgical techniques and postoperative complications management were fundamental in achieving a good result. PMID- 17111807 TI - [Audio-vestibular function in patients with benign paroxysmal positional vertigo]. AB - OBJECTIVE: To investigate the audio-vestibular function and the possible mechanism of benign paroxysmal positional vertigo (BPPV) and to raise the therapeutic strategy. METHODS: Patients with BPPV were tested with pure tone audiometry, high frequency ABR audiometry, bithermal caloric test and vestibular evoked myogenic potential test (VEMP). The positive rate of these otologic function test were analyzed. RESULTS: Primary BPPV comprised 82 percent (70/86) of patients with BPPV. Among all of the patients, the results of pure tone audiometry were abnormal in 52 percent (45/86) of the cases. High frequency auditory brainstem response (ABR) was abnormal in 60 percent (30/50) of cases. Vestibular evoked myogenic potential (VEMP) was abnormal in 34 percent (11/32) of cases who had this examination. And bithermal caloric test were abnormal in 28 percent (20/72) of cases. In the abnormal cases, 67 percent (12/18) of cases were ipsilateral with BPPV. The majority of the BPPV with abnormal results of bithermal caloric test (89%, 16/18) belong to posterior semicircular canal BPPV. CONCLUSIONS: The incidence of primary BPPV was higher than that of secondary BPPV. The abnormality in superior labyrinth was much more correlated with the occurrence of BPPV. The inner ear ischemia might be a factor in the morbidity of BPPV, especially for the primary BPPV. PMID- 17111808 TI - [Relationship between pharyngeal paraesthesia and upper gastrointestinal diseases]. PMID- 17111809 TI - [Computed tomographic and endoscopic analysis of the supraorbital ethmoid cell anatomy]. AB - OBJECTIVE: To understand the anatomy of the supraorbital ethmoid cell and its relationship with the frontal sinus drainage pathway. METHODS: Five patients (5 sides) who had supraorbital ethmoid cell underwent endoscopic frontal sinus surgery. Computed tomographic (CT) scans of the sinuses were obtained in coronal and axial views. The frontal sinus ostium and the supraorbital ethmoid cell were endoscopically identified respectively. RESULTS: On coronal CT scans, the supraorbital ethmoid cell was a separate cell lateral to the frontal sinus. And on axial CT scans, it was lateral and posterior to the frontal sinus drainage pathway. Under endoscope, its opening was lateral and posterior to the frontal sinus ostium. CONCLUSIONS: The supraorbital ethmoid cell extended superolateral the boundaries of the lamina papyracea and the roof of the ethmoid to pneumatize the orbital plate of the frontal bone. PMID- 17111810 TI - [Endoscopic evaluation of mucous membrane inflammation in chronic rhinosinusitis and analysis of correlated factors]. AB - OBJECTIVE: The purpose of this article is to establish an endoscopic score system for quantitative evaluation of the inflammation of mucous membrane in patients with chronic rhinosinusitis (CRS) , and to investigate the correlation of this system with a variety of clinical factors. METHODS: A set of score system was constructed based on anatomic configuration, status of mucous membrane and nasal secretion to evaluate quantificationally the severity of inflammation of CRS. The clinical correlation of this system was studied prospectively in 60 CRS patients, with a variety of clinical factors which included age, duration of disease, previous recurrence and the years from recent recurrence, atopy, serum total immunoglobulin E (TIgE), serum eosinophil cationic protein (ECP), the count of blood eosinophil, the count of tissue inflammatory cell, the extension of CRS indicated by CT, smoking, concomitant chronic inflammation in lower respiratory tract. All above factors were analyzed statistically with the endoscopic score by Pearson correlation and multi-factor linear regression analysis. RESULTS: In pearson analysis, the correlative factors with the evaluated score included age (x1, r = - 0.310, P = 0.016), the extension of disease (x2, r = 0. 810, P < 0.0005), recurrence (x3, r = 0.408, P = 0.001), eosinophil of nasal tissue (x4, r = 0.279, P = 0. 031), duration of disease (x5, r = 0.536, P < 0.0005), concurrent nasal polyps (r = 0.549, P < 0.0005), plasm cell (r = 0. 317, P = 0.014) and years from the recent recurrence (r = 0.385, P = 0.002). In multi-factor linear regression, the five independent predictive factors were recurrence, age, extension of disease, tissue eosinophils, years of disease. The regressive equation is y = 10.148 - 0.152 (x1) + 2.250 (x2) + 3.348 (x3) + 1.233 (x4) + 0.270 (x5). CONCLUSIONS: Appropriate score system by nasal endoscopy is feasible to evaluate quantificationally the degree of inflammation of CRS; being appropriately modified, it is even able to reveal the underlying histological behavior finely. PMID- 17111811 TI - [Clinical features and surgery in children with plastic bronchitis]. AB - OBJECTIVE: To review the clinical features and therapeutic experience in children with plastic bronchitis. METHODS: Fourteen children with plastic bronchitis were reviewed retrospectively, 12 of which were under two years old. The clinical features are characterized by sudden onset, episodes of profound hypoxia and respiratory tract obstruction. SaO2 was between 0.70 and 0.80 even with mask oxygen inhalation. Eight cases were pyretic, 4 cases expectorated jel-like bronchial casts. The chest X-ray picture showed patchy consolidation or atelectasis unilaterally (10 cases) or bilaterally (2 cases). Pulmonary marking thickening and patchy shadow were observed in 2 cases. Twelve cases underwent rigid bronchoscopy and the bronchial casts were removed. Two cases underwent endotracheal intubation. RESULTS: Eight cases of 12 children received therapeutic bronchoscopy were cured. Other 4 cases had second therapeutic bronchoscopy and bronchial casts were removed again in 3 cases, one died from pulmonary hemorrhage. Two cases who underwent endotracheal intubation died from the multiple organ failure (MOF). Pathologic results showed:the bronchial casts were composed mainly of mucus and fibrin, inflammatory cell infiltrate were observed in 6 cases (Type 1, inflammatory), no cellular infiltrate occurred in 8 cases (Type 2, acellular). CONCLUSIONS: Plastic bronchitis is a severe and dangerous disease. The branching plastic casts may obstruct part or the entire tracheobronchial, causing respiratory failure. Bronchoscopy and pathologic examination are essential for it's diagnosis and treatment. PMID- 17111812 TI - [Histopathological study on the regularity of pyriform sinus carcinoma invading adjacent tissue and structure]. AB - OBJECTIVE: To explore the regularity invading adjacent tissue of pyriform sinus carcinoma. METHODS: The whole organ serial section of 68 total or partial laryngectomy and hypopharyngectomy specimen of pyriform sinus carcinoma were histopathologically studied. RESULTS: In 68 pyriform sinus carcinoma, invaded ventricular and paraglottic spaces was 63 and 38 cases respectively, the difference of invasive frequency of both spaces was significantly marked (chi2 = 21.37, P < 0.01). Thyroid cartilage had the most invaded frequency of 92.6% (63/68). The all touching and pressing invasion of laryngeal cartilage was 89 times, and infiltrating invasion was 51 times. The invasive frequency of lateral cricoarytenoid muscle, posterior cricoarytenoid one, thyroarytenoid and interarytenoid ones were 63.2% (43/68), 57.4% (39/68), 55.9% (38/68), 51.5% (35/68) respectively. The invasive frequency of cricoarytenoid and cricothyroid joints were 30.9% (21/68), 17.6% (12/68) respectively. The invasive frequency of superior laryngeal nerve was 67.7% (44/65) , and more than that of recurrent laryngeal nerve (18/65, 27.7%). The pyriform sinus medial wall carcinoma was 14 cases, lateral wall carcinoma 18 cases, medial and lateral wall carcinoma 36 cases. The invaded pyriform sinus apex was 34 cases, normal its apex was 26 ones, submucous invasion of its apex was 8 ones. Light lymphocytic invasion was 66.2% (45/68) and seen most in pyriform sinus carcinoma. Submucous and leaping invasion of pyriform sinus carcinoma were 24 and 8 cases. CONCLUSIONS: Intralaryngeal invasion of pyriform sinus carcinoma arose through paraglottic space first. Laryngeal cartilage membrane and their cartilage were anatomical obstacle against cancerous invasion. Lateral cricoarytenoid muscle, posterior cricoarytenoid one, thyroarytenoid and interarytenoid ones were often invaded. Pyriform sinus medial wall carcinoma invaded intralaryngeal structure easily, its lateral wall carcinoma may invade upward, downward and outward along thyroid cartilage interior wall, medial and lateral wall carcinoma may invade intralaryngeal and extralaryngeal structure, and was the most serious lesion. The invasion of pyriform sinus apex is an important sign of pyriform sinus carcinoma spreading downward to inferior and peripheral tissues of hypopharynx. PMID- 17111813 TI - [Role of ion channel Na-K-2Cl and alpha2 Na, K-ATPase in cochlear potassium cycling and auditory function]. AB - OBJECTIVE: To investigate the auditory function and the role of NKCC1 and alpha2 Na, K-ATPase in the potassium recycling of cochlea. METHODS: NKCC1(+/-) / alpha2 Na, K-ATPase(+/-) mice model was established from NKCC1(+/-) and alpha2 Na, K ATPase(+/-) mice. The auditory function of all strain mice were detected by auditory brainstem response (ABR) and endocochlear potential (EP) to investigate the role of NKCC1 and alpha2 Na, K-ATPase in the potassium recycling of cochlea. Furosemide and ouabain were applied to block the two channels in Castel mice line which can long-time maintain normal auditory function and then their auditory function was detected by ABR to authenticate the mode of potassium recycling in vivo and the relationship between cochlear potassium recycling and NKCC1(+/-) and alpha2 Na, K-ATPase. RESULTS: The mean value for ABR thresholds in response to stimulus was elevated in NKCC1(+/-) and alpha2 Na, K-ATPase (+/-) mice [(38.49 +/ 12.29) dB and (53.32 +/- 7.62) dB) ] respectively, which was significantly increased compared with that observed in wild type mice [(23.13 +/- 3.78) dB, P < 0.05) ]; The EP value of NKCC1(+/-) [(78 +/- 7) mV] and alpha2 Na, K-ATPase(+/-) mice [(71 +/- 14) mV] was decreased compared with that of NKCC1(+/-) / alpha2 Na, K-ATPase(+/-) mice [( 86 +/- 11) mV]. The auditory function of NKCC1(+/-) / alpha2 Na, K-ATPase(+/-) mice could simulate the model of cochlear potassium recycling well. NKCC1 and Na, K-ATPase were great of importance in the potassium recycling, while the two ion channels were in restrict dynamic equilibrium. Castel mice line after administration with furosemide developed significant ABR threshold shifts (P < 0.05) compared with control group. Castel mice line after administration with ouabain also developed greatly significant ABR threshold shifts (P < 0.05) compared with control group. ABR threshold shifts in mice after administration both furosemide and ouabain was attenuated compared with only administration with furosemide (P < 0.01). CONCLUSIONS: Ion channel NKCC1 and alpha2 Na, K-ATPase played important roles in the inner ear potassium recycling. Dysfunction of either of them could influence potassium concentration in the endolymph and lead to hearing loss subsequently. The role of NKCC1 and alpha2 Na, K-ATPase in cochlear potassium recycling was authenticated in vivo. The two ion channels contribute the key role for dynamic equilibrium in cochlear potassium recycling and are of great importance for the metabolism of potassium in the inner ear to maintain the normal auditory function. PMID- 17111814 TI - [Expression and significance of toll like receptor 4 mRNA and nuclear factor kappaB p50 mRNA in human normal nasal mucosa after stimulation by lipopolysaccharide]. AB - OBJECTIVE: To investigate the expression and significance of Toll like receptor (TLR)4 mRNA and nuclear factor (NF)-kappaB p50 mRNA in human normal nasal mucosal cell before and after stimulation by LPS. METHODS: The tissue was obtained from 15 normal middle turbinates (without rhinosinusitis). Every tissue was cultured in vitro, divided into 2 specimens. LPS was added into 15 specimens as LPS group and not added into other 15 specimens as control group. The pathomorphological characters of nasal mucosal cells were observed under optical microscope after stimulation by LPS. The expression of TLR4 mRNA and NF-kappaB p50 mRNA in normal human nasal mucosal cells were evaluated by in situ hybridization. RESULTS: Normal mucociliary agglutinated and mucosal cells were enlarged after stimulation by LPS; The expression of TLR4 mRNA in LPS group was higher than control group obviously. Their average density of light was 1.283 +/- 0.027 in LPS group while 0.538 +/- 0.038 in control group, and there was statistical significance between the two groups (t = 1.761, P < 0.05). The expression of NF-kappaB p50 mRNA was higher than control group obviously, and expressed in cellular nucleus predominantly. Their average density of light was 1. 668 +/- 0.037 in LPS group while 0. 372 +/- 0.052 in control group, and there was statistical significance between the two groups (t = 2. 624, P < 0. 01). CONCLUSIONS: LPS can activate the NF-kappaB p50 of human nasal mucosal cells through TLR4, and it may play some roles in stimulating and damage effect induced by LPS in nasal mucosal cells. PMID- 17111815 TI - [Micro-focus X-ray for cochlear implantation research on small animals]. AB - OBJECTIVE: To develop an imaging system suitable for cochlear implant research on small laboratory animals. METHODS: A novel micro-focus X-ray imaging system was developed. The key features of the system were the use of a micro-focus X-ray source (less than 10 micrometers) and an appropriate choice of source-object and object-image distance. The new X-ray modality was evaluated on heads of cats, guinea pigs and rats. RESULTS: It proved to be an useful tool to provide excellent image of small animal cochlea, allowing a clear electrode position to be established, and also helped greatly in detecting the breakage of electrode array. CONCLUSIONS: Experimental studies performed in cats, guinea pigs and rats confirmed that micro-focus radiography was an important tool for cochlear implant research in laboratory PMID- 17111816 TI - [Tracheoesophageal convex-shunt mode speech reconstruction after total laryngectomy]. PMID- 17111817 TI - [Telomeric loss and anaphase chromosomal bridge in laryngeal squamous cell carcinomas]. PMID- 17111818 TI - [One case report of synovial sarcomas]. PMID- 17111819 TI - [One case report of Langerhans' cell histiocytosis in maxillary sinus]. PMID- 17111820 TI - [Discussion on "Complication of the standard tracheostomy in different conditions"]. PMID- 17111821 TI - [Aetiology and management of vocal nodule in children]. PMID- 17111822 TI - [Control of nasal obstruction in patients with persistent allergic rhinitis]. PMID- 17111823 TI - [Advances in the study of HIV-1 integrase inhibitor of aryl beta-diketoacids]. PMID- 17111824 TI - [Triplex-forming oligonucleotide inhibits the expression of tissue factor gene in endothelial cells induced by the blood flow shear stress in rats]. AB - AIM: To study the effect of antiparallel phosphorothioate triplex-forming oligonucleotide (apsTFO) matching with the shear stress response element (SSRE) of tissue factor (TF) gene promoter region on the expression of TF in endothelial cells (ECs) of rat common carotid artery stenosis. METHODS: The model of common carotid artery middle segment stenosis was established by silica gel pipe loop ligation in SD rats. The mRNA expression and protein synthesis of TF, early growth response-1 (Egr-1) and specificity protein 1 (Sp1) were measured by in situ hybridization (ISH) and immunohistochemistry (IHC) technique. GT21-apsTFO, GT20-apsTFO, GT20-psTFO and FITC-labeled apsTFO, matching with the SSRE of TF gene promoter region, were designed, and intravenously injected into rats at 0.5 h before operation. TFO was detected 4 h after the operation, and the mRNA expression and protein synthesis of TF, Egr-1 and Sp1 were detected 6 h after the operation. RESULTS: There were much fluorescence in vascular tissue, especially in the nuclear of ECs 4.5 h after the injection of apsTFO. The mRNA expression and protein synthesis of TF reduced by 22% - 23% with injection of GT20-apsTFO 6.5 h after stenosis (P < 0.01) and by 10% - 11% with GT21-apsTFO at the same time (P < 0.05). The inhibition by GT20-apsTFO was stronger than that of the GT21 apsTFO (P < 0.05). The expression of TF was not inhibited by the GT20-psTFO (P > 0.05). The mRNA expression and protein synthesis of Egr-1 and Sp1 did not change in the rat treated with GT20-apsTFO, GT20-psTFO and GT21-apsTFO (P > 0.05). CONCLUSION: apsTFO could mero-inhibit the expression of TF gene but could not change the expression of Egr-1 and Sp1 protein. PMID- 17111825 TI - [Effect and mechanism of recombinant hirudin on fibrinolysis]. AB - AIM: To study the effect of recombinant hirudin (rH) on tPA-induced fibrinolysis and the possible mechanism of its action. METHODS: The effect of rH on thrombin fibrin complex (Th-Fn) was detected by 99mTc labeled rH. In the in vitro clot lysis, tPA as plasminogen activator, and recalcified plasma as plasminogen resource were used to study the influence of rH on fibrinolysis by detecting TAFIa, D-Dimer and FXIII. RESULTS: In a canine model of femoral artery thrombosis, a clear radioactivity strip was imaged in 30 - 60 min on a part image, and the femoral vein thrombosis developed at 30 min. rH efficiently inhibited clot regeneration. Addition of TM could inhibit clot lysis obviously, and CPI could shorten the delay of clot lysis which due to TAFIa. There was a dose-dependent relationship with TM concentration and TAFI activation. FXIII activation was inhibited by low concentration of rH ( < or = 0.2 u x mL(-1)), and the level of fibrinolysis product, D-Dimer, increased. CONCLUSION: rH could inhibit the thrombin binding to fibrin. rH inhibited the activation of TAFI and FXIII by combining with thrombin which resulted in enhancement of thrombolysis. PMID- 17111826 TI - [The roles of c-Jun and CBP in the inhibitory effect of quercetin on prostate cancer cells]. AB - AIM: To further uncover the possible mechanism of quercetin-mediated inhibitory effect on prostate cancer cells. METHODS: The cell extracts treated with quercetin or without treatment were used for checking protein expression levels of c-Jun and cAMP response element binding protein (CREB)-binding protein (CBP) by Western blotting assay. Regulatory effects of c-Jun and CBP on the function of androgen receptor (AR) were examined by cotransfection experiment. Finally, a physical interaction of c-Jun and the AR was investigated by coimmunoprecipitation. RESULTS: Quercetin dramatically induced the protein expression of c-Jun which in turn inhibited the AR function. Meanwhile, quercetin had no detectable effect on CBP expression, and the results of transient transfection demonstrated that the ectopic CBP stimulated the transcriptional activity of AR, whereas CBP-mediated stimulation could be attenuated by quercetin. Furthermore, physical interaction of c-Jun and the AR was confirmed by coimmunoprecipitation result. CONCLUSION: Overexpression of c-Jun induced by quercetin had inhibitory effect on the function of AR protein, and increased CBP expression did not reverse the inhibition by quercetin. Together, quercetin mediated inhibition on the AR function might be not by competition with limited amount of CBP in the cell, but through a direct association of c-Jun and the AR. PMID- 17111827 TI - [Synthesis and bioactivity of 2-arylimino-4-thiazolidones]. AB - AIM: To synthesize a series of 2-arylimino-4-thiazolidone derivatives and 2 imidazolino [2,3-b]-4-thiazolidone in order to get some novel potent compounds with nitric oxide synthases (NOS) inhibitory activity. METHODS: The target compounds were prepared by reaction of N-chloroacetyl-1,2,3,4 tetrahydroisoquinoline or N-chloroacetylphthalimide with substituted thioureas, their NOS inhibitory activity were measured. RESULTS AND CONCLUSION: The 15 new compounds were synthesized and most of the reaction yields were over 65%. The structures of new compounds were identified by IR, 1H NMR, MS and elemental analyses. Bioassay indicated that, most of 15 new compounds synthesized had confirmed bioactivities inhibition against NOS. PMID- 17111828 TI - Three new belamcandaquinones from Ardisia punctata. AB - AIM: To study the chemical constituents of Ardisia punctata. METHODS: Compounds were separated with a combination of multi-chromatography. Their chemical structures were determined on the basis of spectral analysis and single crystal X ray diffraction. RESULTS: Three compounds were isolated from chloroform extract of the roots of Ardisia punctata. Their structures were elucidated as 2-tridecyl 3-[(2-tridecyl-3-acetoxy-4-methoxy-6-hydroxy) -phenyl]-6-methoxy-1, 4 benzoquinone (1), 2-tridecyl-3-[(2-tridecyl-4,6-dihydroxy) -phenyl]-6-methoxy-1,4 benzoquinone (2) and 2-tridecyl-3-[(2-pentadecyl-4,6-dihydroxyl) -phenyl]-6 methoxy-,4-benzoquinone (3). CONCLUSION: The three compounds are new 1,4 benzoquinone derivatives. PMID- 17111829 TI - [A new triterpenoid saponin from stems of Akebia trifoliata]. AB - AIM: To study the chemical constituents of the stems of Akebia trifoliata (Thunb.) Koidz. var. australis (Diels) Rehd. METHODS: The compounds were isolated and purified by recrystallization and chromatography on silica gel and ODS column. Their structures were elucidated on the basis of physicochemical properties and spectral analysis. RESULTS: Six compounds were identified as 2 alpha, 3 beta,23-trihydroxyolean-12-en-28-oic acid O-beta-D-xylopyranosyl-(1 --> 3)-O-alpha-L-rhamnopyranosyl-(1 --> 4)-O-beta-D-glucopyranosyl-(1 --> 6)-beta-D glucopyranosyl ester (I), 2 alpha, 3 beta, 23-trihydroxyurs-12-en-28-oic acid O beta-D-xylopyranosyl-(1 --> 3)-O-alpha-L-rhamnopyranosyl-(1 --> 4)-O-beta-D glucopyranosyl-(1 --> 6 )-beta-D-glucopyranosyl ester (II), 2 alpha, 3 beta, 23 trihydroxyolean-12-en-28-oic acid O-alpha-L-rhamnopyranosyl-(1 --> 4)-O-beta-D glucopyranosyl-(1 --> 6)-beta-D-glucopyranosyl ester (III), 2alpha, 3beta, 23 trihydroxyurs-12-en-28-oic acid O-alpha-L-rhamnopyranosyl-(1 --> 4)-O-beta-D glucopyranosyl-(1 --> 6)-beta-D-glucopyranosyl ester (IV), 2 alpha, 3 beta, 23 trihydroxyolean-12-en-28-oic acid O-beta-D-glucopyranosyl ester (V), 2 alpha, 3 alpha, 23-trihydroxyurs-12-en-28-oic acid O-beta-D-glucopyranosyl ester (VI). CONCLUSION: Compound II, named mutongsaponin C, is a new triterpenoid saponin and others were isolated from the stems of Akebia trifoliata (Thunb.) Koidz. var. australis (Diels) Rehd for the first time. PMID- 17111830 TI - [SNP marker and allele-specific diagnostic PCR for authenticating herbs of Perilla]. AB - AIM: To authenticate all the varieties of Perilla (single-species genus), to analyze sequences of rDNA ITS regions and single nucleotide polymorphism (SNP) within them and based on these, to design allele-specific diagnostic PCR primers. METHODS: The rDNA ITS regions of the perilla varieties were sequenced and analyzed by Clustal X 1.8, MEGA 3.0. Allele-specific diagnostic PCR primers that can authenticate all the perilla varieties were designed based on SNPs loci. RESULTS: The length of rDNA ITS sequences of perilla varieties ranged from 612 to 615 bp in size, including ITS1 (230 -232 bp), 5.8S (179 bp) and ITS2 (203 -204 bp). The GC content is about 61.5% - 61.9%. There is not only SNPs in non-coding region ITS1 and ITS2 (ncSNP), but also three coding SNPs (cSNP) loci in the conservative region of 5.8S. All the SNPs have only two allele loci polymorphism. The cSNP in 5.8S is related to the morphology variation among the varieties. Allele-specific diagnostic PCR primers have been designed according to SNPs loci to authenticate accurately all the seeds and leaves of Perilla varieties. CONCLUSION: SNPs in rDNA ITS region can be used as an effective molecular markers to authenticate all the varieties of Perilla. PMID- 17111831 TI - [Liquid chromatography-tandem electrospray ionization ion trap mass spectrometric assay for the metabolites of jatrorrhizine in rat urine]. AB - AIM: To identify the main metabolites of jatrorrhizine in rat urine. METHODS: The rat urine samples were collected 0 - 72 h after ig 12 mg x kg(-1) jatrorrhizine, then the samples were purified through C18 solid-phase extraction cartridge. The purified samples were analyzed by combining liquid chromatography and tandem electrospray ionization ion trap mass spectrometry (LC-ESI/ITMS(n)). Identification and structural elucidation of the metabolites were performed by comparing the changes in molecular masses, retention-times and full scan MS(n) spectra with those of the parent drug. RESULTS: At least seven phase I metabolites (such as de-methyl, de-hydrogen and hydroxyl metabolites) and eleven phase II metabolites (such as glucuronide conjugates and methyl-conjugates) were identified in rat urine. CONCLUSION: The developed LC-ESI/ITMS(n) method is not only simple and rapid but also sensitive and specific for the identification of metabolites of jatrorrhizine in rat urine. PMID- 17111832 TI - [Rapid identification of compounds in cortex moutan by liquid chromatography tandem mass spectrometry]. AB - AIM: To analyze the chemical constituents of cortex moutan by liquid chromatography coupled with electrospray ionization mass spectrometry. METHODS: An on-line optimized HPLC-DAD/MS2 technique was employed. RESULTS: In the negative ion detection mode, 38 components such as monoterpene glucosides, galloylglucoses and acetophenones were isolated. Among them, over thirty compounds were identified, including paeonol, paeonilflorin, oxypaeoniflorin, benzoylpaeoniflorin, benzoyloxypaeoniflorin, galloylpaeoniflorin, galloyloxypaeoniflorin, mundanpioside A, C, D, E, H, etc. by the high performance liquid chromatography with diode-array detection in parallel with electrospray ionization and quadrupole-time of flight tandem mass spectrometry (HPLC-DAD/ESI MS2). CONCLUSION: This method can be used to rapidly determine the constituents of cortex moutan. PMID- 17111833 TI - [The digital criterion of the potential information characteristics of the traditional Chinese medicine chromatographic fingerprints]. AB - AIM: To exploit the characteristic digital criterion for the potential information characteristics of traditional Chinese medicine chromatographic fingerprints, the 37 parameters such as F and I were firstly proposed to disclose the potential information characteristics of traditional Chinese medicine fingerprints. METHODS: The HPLC fingerprints of the Ginkgo biloba extract (GBE) , Ginkgo leaf extract and diphyridamole injection (GLEDI), Ixeris sonchifolia Hance (ISH) and Ixeris sonchifolia Hance injection (ISHI) were compared each other. RESULTS: As far as the peak signal intensity, the uniform of peak signal, resolution and the fingerprint information were concerned. The GBE fingerprint was better than the GLEDI's, and the ISH fingerprint was also better than the ISHI's, then GBE fingerprint was close to the ISHI' s. CONCLUSION: The 37 parameters such as F and I can be used to objectively, authentically and thoroughly display the potential information characteristics of the traditional Chinese medicine chromatographic fingerprints. PMID- 17111834 TI - [Separation and identification of Taxol in the crude extracts of Taxus cuspidata and its callus culture with HPLC-ESI-MS/MS]. AB - AIM: To study the MS/MS fragmentation mechanism of Taxol, and based on it to establish HPLC-ESI-MS/MS technique to separate and identify Taxol in the crude extracts of Taxus cuspidata and its callus culture, consequently to provide a fast and credible method for the analysis of Taxol in natural products. METHODS: Optimized the HPLC-ESI-MS/MS parameters for the sample analysis, and then discussed the ionization and cleavage mechanism of Taxol in ESI-MS and ESI-MS/MS, finally identified the Taxol in the samples with retention time, molecular weight and MS/MS spectra. RESULTS: Elucidated the MS/MS fragmentation mechanism of Taxol, and developed HPLC-ESI-MS/MS method to analyze Taxol in the two samples. CONCLUSION: The HPLC-ESI-MS/MS method is rapid, highly sensitive and specific, so it is suitable for the separation and identification of Taxol in natural products. PMID- 17111835 TI - [Preparation of paclitaxel-loaded chitosan polymeric micelles and influence of surface charges on their tissue biodistribution in mice]. AB - AIM: To prepare paclitaxel-loaded cationic chitosan micelles (PTX-CCM) and paclitaxel-loaded anionic chitosan micelles (PTX-ACM) and study the influence of surface charges on the biodistribution of paclitaxel-loaded chitosan polymeric micelles in mice. METHODS: PTX-CCM and PTX-ACM were prepared by dialysis method and were administered to mice by caudal vein at a dose of 20 mg x kg(-1) body weight. The RP-HPLC method was established to determine the PTX concentrations in the plasma and other tissues of mice. The tissues distribution of PTX-CCM and PTX ACM were evaluated by the pharmacokinetic parameters (AUC, MRT). RESULTS: The diameter and zeta potential of PTX-CCM were 164 nm and +23.7 mV, while those of PTX-ACM were 180 nm and -28.0 mV, respectively. The drug loading and drug encapsulation efficiency for PTX-CCM were 26.4% (w/w) and 76.2% , while those of PTX-ACM were 34.6% (w/w) and 89.9%, respectively. The highest uptake of PTX-CCM and PTX-ACM in liver were 64.72% and 91.84% of dose, respectively. Meanwhile, MRT of both were 5.50 h and 51.39 h prolonged. The highest uptake of PTX-CCM and PTX ACM in spleen were 7.08% and 5.16% of dose, respectively. Meanwhile, MRT of both were 9.04 h and 26.82 h. For PTX-CCM group, the AUC and C(max) of PTX in the lung were 2.71 times and 5.87 times of those of PTX-ACM group respectively. While in both PTX-CCM and PTX-ACM groups, the highest uptake of PTX in the heart were only 0.36% and 0.24% of dose, respectively and PTX in the kidney were only 0.75% and 0.33% of dose respectively. CONCLUSION: PTX-CCM and PTX-ACM showed excellent drug loading capabilities with amount of cationic charges and anionic charges on their surface, respectively. Both PTX-CCM and PTX-ACM groups showed a higher targeting efficiency in the liver and spleen in vivo and accumulated in both tissues for relatively long time, especially in PTX-ACM group. In contrast to PTX-ACM, PTX CCM showed a higher lung targeting efficiency in vivo while PTX-ACM had a stronger retention ability in the lung. Meanwhile in both groups the levels of PTX in the heart and kidney tissues were significantly lower which might decrease the side effects of PTX. PMID- 17111836 TI - [Drug release mechanism of famotidine time-controlled release pellets]. AB - AIM: To study the drug release mechanism of famotidine time-controlled release pellets and to explore the mechanism of "organic acid-induced type drug delivery system". METHODS: The effects of dissociated and undissociated forms of succinic acid on the drug release behavior of famotidine time-controlled release pellets were studied from the following aspects: ion-exchange reaction, hydration, etc. RESULTS: The dissociated succinic acid created new ionic circumstances by ion exchange reaction with Eudragit RS100. Whereas undissociated succinic acid increased the flexibility of the film by distribution in the hydrophobic segment of Eudragit RS100. Effects of both forms of the succinic acid could improve the hydration of Eudragit RS film. As a result, the permeability of the film was improved evidently. CONCLUSION: The lag time of famotidine time-controlled release pellets is induced by the hydrophobicity of the film. After water dissolve the organic acid, the dissociated and undissociated forms of succinic acid interact with the film through different ways. These interactions can change the structure of the film. Therefore the permeability of the film will be improved markedly. PMID- 17111837 TI - [Penetration kinetics of xanthotoxin across human skin and stratum corneum]. AB - AIM: To investigate the penetration kinetics of xanthotoxin in human skin and stratum corneum. METHODS: The penetration experiments were accomplished by the deposit of ethanolic xanthotoxin solution onto human skin and stratum corneum mounted on Franz cells. The diffused xanthotoxin in the receptor solution (1.4% human serum albumin) and the retained amount in the skin and in the stratum corneum after 24 h exposure were quantified by using high performance liquid chromatography. RESULTS: Xanthotoxin flux was increased with the concentration deposited onto the human skin, and when the concentration is above 2.5 mg x mL( 1), there is no influence on the xanthotoxin flux. Similar results were obtained from the stratum corneum. And the peak time for the flux in the stratum corneum was preceded about 6 h earlier than that of the whole human skin. The retained xanthotoxin amount after 24 h exposure in the skin and in the stratum corneum increased according to the concentration deposited and has the tendency to saturate. The lag time of ethanolic xanthotoxin solution in the whole human skin is significantly higher than that in the stratum corneum (P < 0.05). CONCLUSION: The characteristics of penetration kinetics of xanthotoxin will provide the information for concentration choice of topical formulation and give a reference for ultra violet A (UVA) irradiation time confirmation. PMID- 17111838 TI - [The effect of tetramethylpyrazine on the pharmacokinetics of intragastrically administered cyclosporine A in rats]. AB - AIM: To investigate the possible effect of tetramethylpyrazine (TMP), an active ingredient of a commonly used Chinese herb, on the pharmacokinetics of cyclosporine A (CsA) by intragastric administration in rats. METHODS: Forty male Sprague-Dawley rats were equally divided into four groups by randomized block design according to weight. On the first day, after each fasting rat was intragastrically administered CsA (10 mg x kg(-1)), blood samples (0.2 - 0.25 mL) were collected from the tail vein at 0, 1, 2, 3, 4, 6, 8, 12, 24, 36 and 48 h. From day 4 to day 8, each group began to undergo different pretreatments with intragastric administration of water, verapamil (Ver), low and high dose TMP, separately. On day 9, each group intragastrically co-administered CsA (10 mg x kg(-1)) and different pretreatment compounds mentioned above, then blood samples were collected according to the schedule of the first day. The whole blood concentration of CsA was determined by HPLC. Main pharmacokinetic parameters were calculated and compared by statistic analysis. RESULTS: In the group of water pretreated and co-administrated with CsA, no significantly different pharmacokinetic parameters of CsA were found. After Ver pretreatment and co administration with CsA, AUC(0-48 h) and C(max) were increased significantly (P < 0.01 and P < 0.05); T(1/2) beta and CL were markedly prolonged and decreased (P < 0.05); T(max) and V were not apparently influenced. After low dose TMP pretreatment and co-administration with CsA, there was no significant difference in the pharmacokinetic parameters of CsA, in spite of the increasing trends of AUC(0-48 h) and C(max). After high dose TMP pretreatment and co-administration with CsA, AUC(0-48 h) and C(max) of CsA were increased significantly (P < 0.01), but there was no significant change in other parameters. CONCLUSION: It was indicated that the high dose of TMP could apparently increase the intragastric absorption extent of CsA, while almost had no effect on its elimination process. PMID- 17111839 TI - [Molecular simulations of the loading of methadone and buprenorphine into carbon nanotubes]. AB - AIM: To simulate the inhalation of the C21H27NO and C29H41NO4 molecules, the effective components of methadone and buprenorphine, into carbon nanotubes, and discuss the feasibility of the loading of methadone and buprenorphine into carbon nanotubes. METHODS: The MM + force-field based molecular dynamics (MD) method uas used. RESULTS: The ends-opened carbon nanotubes with diameter larger than 1 or 1.25 nm can initiatively inhale the C21H27 NO or C29H41NO4 molecule, and both two molecules have higher potential energy at the open ends of the carbon tubes than that at the middle of the tubes; the present single-walled nanotubes are very suitable for the loading of methadone and buprenorphine. CONCLUSION: It is possible to make sustained-release detoxification agents with methadone- or buprenorphine-loaded carbon nanotubes. PMID- 17111840 TI - [Bioequivalence assessment of pioglitazone hydrochloride oral preparation by limited sampling strategy]. AB - AIM: To develop limited sampling strategy (LSS) for estimation of C(max) and AUC(0-t) and assessing the bioequivalence of two pioglitazone hydrochloride (PGT) preparations. METHODS: Healthy subjects (n = 20), enrolled in a bioequivalence study, were received 30 mg PGT po of reference or test formulation. The plasma concentration of PGT was determined by the validated HPLC method. A multiple linear regression analysis of the Cmax and AUC(0-t) against the PGT concentration for the reference formulation was carried out to develop LSS models to estimate these parameters. The models were internally validated by the Jackknife method and externally validated using simulated sets generated by Monte Carlo method. The best model was employed to assess bioequivalence of the two PGT formulations. RESULTS: The linear relationship between pharmacokinetics parameters and single concentration point was poor. Several models for these parameters estimation met the predefined criteria (r2 > 0.9). The Jackknife validation procedure revealed that LSS models based on two sampling times (C1, C2.5 and C1.5, C2.5 for C(max); C1.5, C9 and C2.5, C9 for AUC(0-t) predict accurately. Mean prediction errors (MPE) were less than 3%, and mean absolute prediction error (MAE) were less than 9%. The prediction error (PE) beyond 20% was less than 5% of total samples. Model external validation by Monte Carlo simulated data indicated that the most informative sampling combinations were C1.5, C2.5 for C(max), and C1.5, C9 for AUC(0-t), respectively. MPE and MAE of the proposed models were less than 5% , and 9% respectively. The PE beyond 20% was less than 5% of the total. Bioequivalence assessment of the two PGT formulations, based on the best LSS models, provided results similar to those obtained using all the observed concentration-time data points, and indicated that the two PGT formulations were bioequivalent. CONCLUSION: The LSS method for bioequivalence assessment of PGT formulations was established and proved to be applicable and accurate. Thus, it could be considered appropriate for PGT bioequivalence study with inexpensive cost of sampling acquisition and analysis. Key words: pioglitazone hydrochloride; limited sampling strategy; Monte Carlo simulation; bioequivalence PMID- 17111841 TI - [Echogenic phospholipids-based gas-filled microbubbles as delivery system of antisense oligodeoxynucleotides]. AB - AIM: To investigate the feasibility of transfer antisense oligodeoxynucleotides (AS-ODNs) by the phospholipids-based gas-filled microbubbles (PGM) under ultrasound activation. METHODS: An antisense oligodeoxynucleotides sequence ZL combined with luciferase reporter plasmid was used. A breast cancer cell line SK BR-3 was exposed to different conditions to investigate the effects of such factors as ZL concentration, PGM concentration, mechanical index (MI) and ultrasound exposure duration on transfection efficiency and cell viability. The transfection efficiency and cell viability by other lipid vectors such as lipofectamine and liposome were also tested, whose results were comparied with that of PGM. Transfection efficiency was detected by fluorescence microscopy. Cell viability was verified by PI (propidium iodide) assay. RESULTS: Among the factors tested, ultrasound exposure duration, MI and PGM concentration had obvious impacts on transfection efficiency and cell viability. The results showed that the optimal ultrasound condition was the exposure to ultrasound at MI 1.0 for 30 s with 2% PGM concentration, which gave an overall transfection efficiency of 78% +/- 10%, increased nearly 18 folds over the transfection by PGM (4.0%) or lipofectamine (4.3%) without ultrasound. Under same ultrasound conditions, different vectors showed significant difference in transfection efficiency while there are similar results in cell viability. CONCLUSION: Under proper ultrasound conditions, PGM can markedly enhance AS-ODNs transfection efficiency. PMID- 17111842 TI - [The protective effects of ginsenoside RG1 and Rb1 against damage of HaCaT cells by ultraviolet B]. AB - AIM: To investigate the survival rate and the level of HaCaT cells damage with ultraviolet B (UVB) radiation at various doses, and observe the protective effects of ginsenoside Rg1 and Rb1 in vitro. METHODS: MTT assay was employed to analyze the cell survival rate after UVB radiation of 30, 60, 90 and 120 mJ x cm( 2). The damage of nucleolus and the protective effects of ginsenoside Rg1 and Rb1 were scanned by Hoechst 33258 staining and single cell gel electrophoresis assay (SCGE). RESULTS: It was found that the cell survival rate decreased gradually and the damage of nucleolus aggravated as the radiation dose increased from 30 mJ x cm(-2) to 120 mJ x cm(-2). At the dose of 20 microg x mL(1-), obvious protective effect of ginsenoside Rg1 and Rb1 can be observed against UVB radiation-induced HaCaT cells growth inhibition and nucleolus damage. CONCLUSION: UVB radiation inhibits HaCaT human keratinocytes growth and ginsenoside Rg1 and Rb1 can relief the damage. PMID- 17111843 TI - [Determination in vitro of rat plasma protein binding rate of sinomenine by using microdialysis method]. AB - AIM: To determine in vitro the rat plasma protein binding rate by using microdialysis method. METHODS: The binding rate was determined by using microdialysis probe as sampling tools and zero-net flux method as calibrating method. The regression equation was made by the difference of concentrations between the dialysis sample and the perfusate. The x-intercept of regression equation was the free drug concentration (Cf). The plasma protein binding rate was calculated by using the following equation: f = ( C0 - Cf)/C0. RESULT: The binding rate was kept relatively stable in the studied concentration range. CONCLUSION: It is feasible that the plasma protein binding rate can be determined by using microdialysis method. PMID- 17111844 TI - [Malignant lyphomas/non-Hodgkin lymphomas. Chief symptoms: lymph node swelling, occasionally B-symptoms]. PMID- 17111845 TI - [What is your diagnosis? AV-node reentrant tachycardia, AV block I]. PMID- 17111846 TI - [Hereditary pancreatitis]. AB - Hereditary pancreatitis should be assumed if other risk factors for the disease can not been identified and if the patient has a family history for recurrent pancreatitis, chronic pancreatitis or pancreatic cancer. Since patients with chronic pancreatitis due to mutations in the cationic trypsinogen-gene have a much higher lifetime risk of developing pancreatic cancer, specifically if they are smokers, an adequate long-term follow up in specialized centers is recommended. The most frequent genetic changes in patients with hereditary pancreatitis are mutations in the cationic trypsinogen gene. Mutations in the CFTR-gene or SPINK1-gene have been reported in patients with idiopathic pancreatitis. The clinical relevance and the therapeutic consequences of these mutations is still controversial. Genetic testing is recommended when a patient with idiopathic pancreatitis is under 25 years at diagnosis or when one or more family members have either pancreatitis or pancreatic cancer. Genetic analysis of asymptomatic family members should only be offered after adequate genetic counselling. Prenatal diagnostic is not recommended. PMID- 17111847 TI - [Nutrition, probiotics, antibiotics, antioxidative therapy, endoscopy in chronic pancreatitis]. AB - Treatment of chronic pancreatitis is dependent on the stage of the disease and consists of several arms: treatment of pain when ever possible according to its pathogenesis; treatment of complications primarily by interventional endoscopy, in cases of failure by surgery; therapy of exocrine insufficiency with porcine pancreatic extracts; treatment of endocrine insufficiency with insulin. Pseudocysts can be drained according to their location by either the transgastric, transduodenal, transpapillary or transcutaneous route. Distal prepapillary stenoses of the main pancreatic duct can be handled by placement of a plastic stent; similarily to treatment of biliary strictures. Stones leading to obstruction of the main pancreatic duct can be disintegrated by extracorporeal shock wave lithotripsy (ESWL) and the fragments removed by endoscopy after papillotomy. Transgastral endoscopic drainage of retroperitoneal necroses is still experimental. Prospective randomized multicenter trials comparing surgery with interventional endoscopy are still lacking. Failure of endoscopic therapy or suspicion of tumor is clearly an indication for surgery. There is no need for a specific diet in patients with chronic pancreatitis without having diabetes. In severe attacks, clinically similar to acute pancreatitis, enteral nutrition via a jejunal tube is replacing parenteral nutrition. However, prospective comparative trials are still mandatory. Prophylactic application of antibiotics in patients with pancreatic necrosis is again under debate. Whether probiotics are capable to decrease the risk of secondary pancreatic infection of necrosis has not been thoroughly studied. The hypothesis that capture of oxygen free radicals by drugs such as selenium may prevent frequency and severity of acute relapses has also not been proven. PMID- 17111848 TI - [More quality of life again with tolterodine]. PMID- 17111849 TI - [Klatskin tumors--diagnostic and interventional therapy]. AB - Klatskin tumors are defined as malignant tumors of the bile duct involving the bifurcation and intrahepatic bile ducts. The most common presenting clinical feature, obstructive jaundice, usually occurs with advanced disease. Diagnostic tools currently available are therefore either performed too late or are not able to detect early disease stage. Imaging procedures for diagnosis and staging are ultrasonography, magnetic resonance imaging with cholangiopancreaticography, intravenous bolus-enhanced spiral computed tomography and endoscopic retrograde cholangiopancreaticography. Before initiating any palliative measure, a proper staging and a surgical consultation at a hepatobiliary center is necessary. To assess resectability, additional diagnostic methods like angiography, positron emission tomography, cholangioscopy, endoscopic or intraluminal ultrasonography and finally even explorative laparoscopy may be required. At time of diagnosis only a small percentage of Klatskin tumors is curative resectable. Therefore, palliative treatment options play an important role. Endoprostheses insertion is the method of choice to relieve jaundice. Although it improves quality of life, it does not seem to improve survival time. Definitive evidence for a benefit of additional radio and/or chemotherapy is still missing. Photodynamic therapy, a light therapy, is the first approach leading to an improvement of cholestasis and quality of life as well as to a prolongation of survival time. PDT should therefore be offered to all patients with nonresectable cholangiocarcinoma. PMID- 17111850 TI - [Klatskin tumor surgical treatment]. AB - Due to the difficulties of diagnosis of central bile duct carcinoma an experienced surgeon should be involved in the decision-making process at an early stage. This is true in particular with regard to the fact that currently the only curative treatment option is an extended bile duct resection with up to 80% five year survival rates. In case of a too small liver volume, contralateral selective percutaneous embolization of the portal vein can be performed. Liver transplantation is reserved to a selected group of patients with local inoperability, but it is becoming more and more important due to the possibility of living donor liver transplantation associated with multimodal treatment strategies. PMID- 17111851 TI - [What is your diagnosis? Pyoderma grangrenosum]. PMID- 17111852 TI - [Knee ligament injuries]. AB - Like no other joint of the human body the knee depends on intact ligaments. Knee instability due to ligament injuries will cause abnormal joint kinematics, and thereby is made responsible for secondary damage to other important knee joint structures. Diagnosis of knee ligament injuries is based on the detailed history with often typical injury patterns, as well as on the physical examination with specific knee ligament tests. In addition radiological evaluation is used. The range of knee ligament injuries is wide. Beginning with an isolated medial collateral ligament rupture which will heal with conservative treatment, they range to knee dislocation, a serious injury which needs emediate care and is associated with a high incidence of complications. Surgical procedures aim to reconstruct knee ligaments as anatomical as possible to provide for a long term stable knee joint. PMID- 17111853 TI - [Invasive meningococcal infections: two cases that demonstrate the broad spectrum in clinical manifestation and outcome]. AB - Invasive meningococcal infections show a broad clinical picture including sepsis and meningitis. Here we report on a case of sepsis and a case of meningitis, two clinical manifestations of meningococcal infections with striking differences in the clinical presentation and outcome. Meningococcal sepsis is characterized by a systemic release of endotoxins, that triggers an intense cytokine response of the host that can lead to shock and multi organ failure and death within hours. Meningococcal meningitis occurs when bacteria breach into the subarachnoidal and ventricular space during bacteremia and mortality is much lower that in sepsis. Thus meningitis may be seen as a consequence of lower pathogenicity and/or more efficient host control of the meningococci compared to sepsis. PMID- 17111854 TI - [Status asthmaticus with rhabdomyolysis of the respiratory muscles]. AB - A 40-year-old roofer called the medical emergency team because of sudden dyspnea and chest constriction. He was suffering from bronchial asthma that exacerbated probably due to the inhalation of a solvent while at work. Because of status asthmaticus he was intubated immediately after arrival in the hospital and breathing had to be assisted artificially for ten hours. After extubation he complained of chest pain. In view of an elevated creatinkinase of 4368 U/1 this was interpreted as rhabdomyolysis of the respiratory muscles due to the status asthmaticus. As a cause of chest pain and elevation of creatinkinase after severe respiratory work under hypoxic conditions rhabdomyolysis of the involved musculature has to be considered. PMID- 17111855 TI - [CME--ultrasound diagnosis 13/solution. Right sided hip pain. Osteolytic metastasis of a renal cell carcinoma]. PMID- 17111861 TI - [Hygiene and wound bacteriology]. PMID- 17111862 TI - [The nurse's role in multiple sclerosis treatment adherence]. PMID- 17111863 TI - [A nursing society in France, a recent old history]. PMID- 17111864 TI - [Creation of the National Society of Nurses, analysis of the legal proposition]. PMID- 17111865 TI - [Committee for inter-society liaison, professional societies and inter relationship]. PMID- 17111866 TI - [Recent reforms of health professions societies]. PMID- 17111867 TI - [Functioning of the social assurance section]. PMID- 17111868 TI - [National Society of Pedicurists-Podiatrists, a chance for the profession]. PMID- 17111869 TI - [Society of Masseurs-Kinesitherapists, an ongoing construction]. PMID- 17111870 TI - [Towards European cooperation in the professional regulation of nursing]. PMID- 17111871 TI - [Society of Nurses in Portugal]. PMID- 17111872 TI - [The professionalization of nursing in the French milieu]. PMID- 17111873 TI - [Nursing societies and globalization]. PMID- 17111874 TI - [Nursing society, a unique voice, professional landmarks]. PMID- 17111875 TI - [Nursing societies and disorder]. PMID- 17111876 TI - [8/10 Lack of information as the origin of serious medical care errors]. PMID- 17111877 TI - [Hodgkin lymphoma (Hodgkin disease)]. PMID- 17111878 TI - [What is your diagnosis? Severe celiac disease (endemic sprue)]. PMID- 17111879 TI - [Pancreatic cancer]. AB - Pancreatic cancer is the fourth leading cause of death due to cancer. The most common cancer in the pancreas is ductal adenocarcinoma. Pancreatic cancer is characterized by alterations in K-Ras, INK4a, Tp53 and SMAD4. Similar to colon cancer a cancer progression model for pancreatic cancer has been proposed. The precursor lesions are called pancreatic intraepithelial neoplasia. Patients with tumors in the head of the pancreas may present deep jaundice without pain. Multidetector CT incorporating dual-phase imaging in the arterial and venous phases of enhancement is the preferred imaging modality for the diagnosis and staging of pancreatic cancer. Gemcitabine is still the standard for unresectable locally advanced disease or distant metastasis. PMID- 17111880 TI - [Intestinal ischemia--non-surgical approach]. AB - Prognosis of acute mesenteric ischemia depends on the quality of the initial management of the treating physician. To reduce the still appallingly high mortality rate of this disease a rapid, invasive and aggressive diagnosis and treatment is necessary. However, early signs and symptoms of mesenteric ischemia as well as laboratory studies are nonspecific. Therefore, definitive diagnosis requires invasive testing using mesenteric angiography. Surgery should not be delayed in patients with peritoneal signs and suspected of having intestinal perforation or gangrene. PMID- 17111881 TI - [Intestinal ischemia--surgeon's view]. AB - In mesenteric ischemia, we are still facing the problem of late diagnosis: Only early start of therapy within the first 12 hours of symptom onset can reduce mortality rates below 50 percent. Most effective diagnostic tests are catheter angiography with the option of therapeutic intervention and multi-slice computed tomography. As for treatment strategy reasons, acute arterial thrombosis and embolic ischemia should be regared separately from venous, non-occlusive (NOMI) and chronic mesenteric ischemia. Acute arterial ischemia is treated by immediate open exploration, revascularization and bowel resection. Venous ischemia is treated best by catheter anticoagulation, while in NOMI restoration of altered hemodynamics and selective vasodilatation is the therapy of choice. Late complications in mesenteric ischemia are malabsorption, bypass graft thrombosis and the small-bowel-syndrom, that can be solved in selected cases by small bowel transplantation. PMID- 17111882 TI - [Toxic megacolon--the position of the internist]. AB - Toxic megacolon is a rare but typical complication of ulcerative colitis. But also every inflammatory bowel disease may be cause of toxic megacolon, including especially Crohn's colitis and pseudomembranous colitis (PMC). There is to mention, that PMC becomes more frequent and more important in the last years. Toxic megacolon is defined by radiological, clinical and labaratory criteria: colonic distension (<6 cm), fever, tachycardia, hypotension, electrolyte disturbances, leucocytosis and anemia. Intensive medical therapy is primarly the therapy of choice, but clinical parameters should be closely monitored. In patients with ulcerative colitis high dosis of corticosteroids are given intravenously. If there is no clinical improvement after 48 to 72 hours, colectomy is required. The use of alternative therapies like cyclosporine or infliximab has to be discussed as possible before toxic megacolon occurs. PMID- 17111883 TI - [Toxic megacolon--surgical point of view]. AB - Toxic megacolon is an infrequent, but potentially fatal complication of a fulminant colitis. Toxic colonic dilatation, also caused by ischaemic or infectious inflammation like pseudomembranous colitis, mostly occur in patients with inflammatory bowel disease. Toxic mega-colon is defined as segmental or total colonic distension of >6 cm with the presence of clinical signs of acute colitis and systemic toxicity. Because of the associated high morbidity and mortality the early diagnosis and the management play an important role. The free perforation means a fourfold increase in the mortalitiy of the acute colitis. Recognition of toxic megacolon is underlaying by x-ray of the abdomen with colonic distension and a lack of haustral pattern. Accompanying distension of the small bowel can predict the development of the disease. CT scanning shows a diffuse wall thickening, pericolic inflammation and abnormal haustral pattern and can also detect subclinical perforation or abscesses. The management of toxic megacolon should be with intravenous parenteral nutrition, adaequate supplementation of intravenous fluids and correction of electrolytes abnormalities and the therapy of colitis with corticosteroids. Antibiotics are indicated in infectious disease or bacteriemia and also in colonic perforation. Surgical intervention is indicated by the onset of signs of progression of the disease and complications as perforation, uncontrollable bleeding or distension. The surgical procedure of choice is colectomy and ileostomy. The mortality and morbidity was decreased by avoiding rectal excision. The rectum is closed as a Hartmann's procedure or a mucous fistula is created. A secondary ileoanal pouch can be created at a later date. The interdisciplinary approach with optimal timing of surgical intervention can decrease the morbidity and mortality of toxic megacolon. PMID- 17111884 TI - Medication options for the treatment of CAP. PMID- 17111885 TI - BMI monitoring in the management of obesity in toddlers. PMID- 17111886 TI - Hormone therapy in postmenopausal and perimenopausal women. PMID- 17111887 TI - Antidepressants in older persons. PMID- 17111888 TI - Interventions for molluscum contagiosum. PMID- 17111889 TI - Otitis externa: Review and clinical update. AB - Otitis externa can take an acute or a chronic form, with the acute form affecting four in 1,000 persons annually and the chronic form affecting 3 to 5 percent of the population. Acute disease commonly results from bacterial (90 percent of cases) or fungal (10 percent of cases) overgrowth in an ear canal subjected to excess moisture or to local trauma. Chronic disease often is part of a more generalized dermatologic or allergic problem. Symptoms of early acute and most chronic disease include pruritus and local discomfort. If left untreated, acute disease can be followed by canal edema, discharge, and pain, and eventually by extra-canal manifestations. Topical application of an acidifying solution is usually adequate in treating early disease. An antimicrobial-containing ototopical is the preferred treatment for later-stage acute disease, and oral antibiotic therapy is reserved for advanced disease or those who are immunocompromised. Preventive measures reduce recurrences and typically involve minimizing ear canal moisture, trauma, or exposure to materials that incite local irritation or contact dermatitis. PMID- 17111890 TI - Seasonal affective disorder. AB - Patients with seasonal affective disorder have episodes of major depression that tend to recur during specific times of the year, usually in winter. Like major depression, seasonal affective disorder probably is underdiagnosed in primary care settings. Although several screening instruments are available, such screening is unlikely to lead to improved outcomes without personalized and detailed attention to individual symptoms. Physicians should be aware of comorbid factors that could signal a need for further assessment. Specifically, some emerging evidence suggests that seasonal affective disorder may be associated with alcoholism and attention-deficit/hyperactivity disorder. Seasonal affective disorder often can be treated with light therapy, which appears to have a low risk of adverse effects. Light therapy is more effective if administered in the morning. It remains unclear whether light is equivalent to drug therapy, whether drug therapy can augment the effects of light therapy, or whether cognitive behavior therapy is a better treatment choice. PMID- 17111891 TI - Nutrition in toddlers. AB - Toddlers make a transition from dependent milk-fed infancy to independent feeding and a typical omnivorous diet. This stage is an important time for physicians to monitor growth using growth charts and body mass index and to make recommendations for healthy eating. Fat and cholesterol restriction should be avoided in children younger than two years. After two years of age, fat should account for 30 percent of total daily calories, with an emphasis on polyunsaturated fats. Toddlers should consume milk or other dairy products two or three times daily, and sweetened beverages should be limited to 4 to 6 ounces of 100 percent juice daily. Vitamin D, calcium, and iron should be supplemented in select toddlers, but the routine use of multivitamins is unnecessary. Food from two of the four food groups should be offered for snacks, and meals should be made up of three of the four groups. Parental modeling is important in developing good dietary habits. No evidence exists that early childhood obesity leads to adult obesity, but physicians should monitor body mass index and make recommendations for healthy eating. The fear of obesity must be carefully balanced with the potential for undernutrition in toddlers. PMID- 17111892 TI - Information from your family doctor: Giving your child the best nutrition. PMID- 17111893 TI - Diagnosis of acute abdominal pain in older patients. AB - Acute abdominal pain is a common presenting complaint in older patients. Presentation may differ from that of the younger patient and is often complicated by coexistent disease, delays in presentation, and physical and social barriers. The physical examination can be misleadingly benign, even with catastrophic conditions such as abdominal aortic aneurysm rupture and mesenteric ischemia. Changes that occur in the biliary system because of aging make older patients vulnerable to acute cholecystitis, the most common indication for surgery in this population. In older patients with appendicitis, the initial diagnosis is correct only one half of the time, and there are increased rates of perforation and mortality when compared with younger patients. Medication use, gallstones, and alcohol use increase the risk of pancreatitis, and advanced age is an indicator of poor prognosis for this disease. Diverticulitis is a common cause of abdominal pain in the older patient; in appropriately selected patients, it may be treated on an outpatient basis with oral antibiotics. Small and large bowel obstructions, usually caused by adhesive disease or malignancy, are more common in the aged and often require surgery. Morbidity and mortality among older patients presenting with acute abdominal pain are high, and these patients often require hospitalization with prompt surgical consultation. PMID- 17111895 TI - Information from your family doctor: Giant cell arteritis and polymyalgia rheumatica: what you should know. PMID- 17111894 TI - Polymyalgia rheumatica and giant cell arteritis. AB - Polymyalgia rheumatica and giant cell arteritis are common, closely related vasculitic conditions that almost exclusively occur in patients older than 50 years. They may be manifestations of the same underlying disease and often coexist. Patients with polymyalgia rheumatica usually present with acute onset of stiffness and pain in the shoulder and pelvic musculature, which may be accompanied by fever, malaise, and weight loss. If untreated, polymyalgia rheumatica may result in significant disability. Giant cell arteritis may manifest as visual loss or diplopia, abnormalities of the temporal artery such as tenderness or decreased pulsation, jaw claudication, and new-onset headaches. Erythrocyte sedimentation rate and temporal artery biopsy help make the diagnosis. Giant cell arteritis requires urgent diagnosis because without treatment it may lead to irreversible blindness. Patients with either condition also may have nonspecific symptoms. Corticosteroids are the mainstay of therapy for both conditions, with higher doses required for treatment of giant cell arteritis. Duration of corticosteroid therapy can be five years or longer before complete clinical remission is achieved. Monitoring for corticosteroid-associated side effects such as osteoporosis and diabetes, as well as for relapses and flare ups, is key to chronic management. The prognosis for either condition, if treated, is good. PMID- 17111896 TI - Information from your family doctor: Erectile dysfunction: what you should know. PMID- 17111897 TI - Information from your family doctor: Prostate cancer: what you should know. PMID- 17111898 TI - Information from your family doctor: Testicular cancer: what to look for. PMID- 17111899 TI - Chronic low back pain. PMID- 17111900 TI - FPIN's clinical inquiries. Psychosocial interventions delivered by primary care physicians to patients with depression. PMID- 17111901 TI - Asymmetry of the thorax and hypoplastic hand. PMID- 17111902 TI - Treatment of patients with literacy issues. PMID- 17111903 TI - HIM issues front and center. PMID- 17111904 TI - The value of personal health records. A joint position statement for consumers of healthcare. PMID- 17111905 TI - Vision 2006. 10 years of advancing professional success, lessons for the decade to come. PMID- 17111906 TI - The HIM impact on EHRs. PMID- 17111907 TI - Ready for the deficit reduction act? PMID- 17111908 TI - What's next for EHR certification? And what product certification means for HIM. PMID- 17111909 TI - E-discovery and HIM. PMID- 17111910 TI - Crimes and sanctions. Current controversies over HIPAA's criminal penalty. PMID- 17111911 TI - Key points of the UB-04. PMID- 17111912 TI - Establishing professional development goals. The means to your future. PMID- 17111913 TI - EHR case study. How a well-managed selection process led to a successful IT implementation. PMID- 17111914 TI - Coding pregnancy. PMID- 17111915 TI - Home care coding basics. Potential PPS changes may affect home care coding. PMID- 17111916 TI - Exploring the edges of the profession. PMID- 17111917 TI - [The technologies of genetic engineering in treatment of chronic lower limb ischemia]. AB - The article contains experimental data on angiogenesis stimulated by plasmid containing the angiogenin gene. After the introduction of the gene construction, the number of capillars in the chorion-allantois membrane increases 2 to 3 times; in an ischemized limb of a rat it increases by 20 to 30%. Intramuscular administration of genetic engineering construction to patients with chronic lower limb ischemia improved the patients' condition, consisting in an increase in painless walking distance and ankle-brachial index, as well as in trophic defect healing and the betterment of muscular perfusion. Positive effects were noted after 2 to 4 weeks of treatment and remained during 6 to 24 months. There were no side-effects, except low grade fever during 1 to 2 days. PMID- 17111918 TI - [The use of modern diagnostic and preventive technologies in children with hereditary and congenital intellectual developmental disorders]. AB - The article is dedicated to the problems of intellectual disorders in children suffering from congenital and hereditary diseases, and reflects the issues of the medicosocial significance of neuropsychical impairment in children and the proportion of ethiological factors in the genesis of mental retardation. The authors consider modern diagnostic and preventive technologies that are used in pediatric practice in children with hereditary and congenital intellectual developmental disorders. PMID- 17111919 TI - [The problem of cerebral stroke: its contemporary state]. AB - The authors consider the prevalence and the clinical outcomes of cerebral stroke in Russia and the most promising methods directed towards lowering stroke incidence and raising the quality of pre-hospital and hospital aid. The authors also pay their attention to rehabilitation techniques improving the life quality of patients after cerebral stroke, and characterize the areas of scientific research into the lowering of the medicosocial consequences of cerebral stroke. PMID- 17111920 TI - [The role of regulatory cells CD4+CD25+ in the development of chronic infective diseases]. AB - Regulatory T-cells CD4+CD25+Foxp3 (Treg) present a small subpopulation of CD4+ T lymphocytes, which develop in the thymus and are disseminated into peripheral lymphoid organs on the 3rd or the 4th day of the neonatal period. Treg play a decisive role in the pathogenesis of autoimmune diseases and the development of tolerance to transplantation antigens, regulate the immune response to allergens, and suppress antimicrobial immunity. Treg suppress proliferation as well as the cytotoxic effect and the secretion of Th1 and Th2 cytokines by effectory T lymphocytes, thus limiting the strength of the immune response of effectory T cells, which makes them impossible to adequately control viral and bacterial infections. Recognition via antigen-presenting cells and the subsequent induction of the proliferation of antigen-reactive T and B lymphocytes, directed towards infectious agent elimination, is accompanied by the activation of regulatory T cells as well, which leads to immune response suppression; repeated microbial infections are not only able to strengthen T-cell immunity by generating memory T cells, but can also strengthen the suppressive activity of endogenous T regulators CD4+CD25+. Moreover, T reg are capable of the direct recognition of a microbial product; these cells selectively express Toll-like receptors (TLR)-2, 4, -5, -7, and -8. Under normal conditions T reg are anergic, but are capable of direct proliferation in response to stimulation by TLR ligands, expressed on microbes and parasites. Treg removal enforces protective immune response to contagious microorganisms, including bacteria, viruses, and fungi, which leads to the elimination of pathogens from the host organism. The removal of Treg population will help to accomplish infectious pathogen elimination and diminish inflammation within a short period of time. PMID- 17111921 TI - [Pharmacological approaches to the therapy of Alzheimer's disease]. AB - The role of apolipoprotein E (ApoE) genotype in the forming of response to various kinds of pathogenetic therapy of Alzheimer's disease (AD) was studied in two samples of patients receiving cholinergic or neurotrophic therapy during four months. The samples were formed by pair control method according to the following parameters: clinical type of AD (presenile or senile) and the severity of dementia (mild or moderate). The average scores according to cognitive scales (MMSE and ADAS-cog) and to the scale which evaluates the everyday activity of patients (IADL) were practically identical. Patients in group I were treated with exelon (rivastigmine) in maximal individually tolerable doses (76.7% of the patients were receiving > or = 6 mg/ day). Patients in group II received two courses of cerebrolysin (20 i.v. injections of 30.0 ml of the drug in 100 ml of normal saline) with an 8-week break. Both treatments had a significant therapeutic effect, but the proportion of responders to different kinds of therapy depended on ApoE genotype. Among ApoEepsilon4(+) patients the proportion of responders was practically equal in both treatment groups (30.8% and 31.2%), while among ApoEepsilon4(-) patients the proportion of responders to cerebrolysin was three times as big as the proportion of responders to exelon (47.0 and 14.3%, respectively). In the cerebrolysin group the proportion of responders was 1.5 times bigger among ApoEepsilon4(-)patients, while in the exelon group, on the contrary, the proportion of responders was twice bigger among ApoEepsilon4(+) patients (31.2% and 14.3%, respectively). Long-term effects of cerebrolysin (two months after the end of the treatment) in ApoEepsilon4(-) patients were also higher than those in ApoEepsilon4(+) patients, while in exelon patients long-term effects did not depend on ApoEepsilon4. The study demonstrates the significance of the detection of ApoEepsilon4 genotype for the choice of a therapeutic approach in patients with mild or moderate dementia caused by AD. PMID- 17111922 TI - [Bacterial translocation under the conditions of acute bowel obstruction]. AB - The article is dedicated to bacterial translocation under the conditions of acute small and large bowel obstruction, studied in experiment using bacteriological methods. The degree of contamination of internal organs, blood, and peritoneal exudate in the dynamics of the pathological process was determined. The study found that the use of allopurinol, a xanthine oxidase inhibitor, reduced the intensity of bacterial translocation. PMID- 17111923 TI - [The condition and prospects of Russian endocrinology]. AB - The article presents a brief essay dedicated to the development of endocrinology in our country since pre-war time. The article covers the most promising areas of endocrinology, from endemic goiter to the introduction of the newest technologies to endocrinology, and informs the readers about structural and organizational changes in the diabetological service, including the establishment of Institute of Diabetes and Institute of Pediatric Endocrinology, the first ones in Russia, the establishment of diabetological service, including State Registry of Diabetic Patients, a school for adult patients, etc. Another issue is the establishment of Growth Center and the introduction of genetic engineering growth hormone preparations into the practice of treatment of short children. Once again, the authors emphasize the issue of fighting endemic goiter. Many tasks of Russian endocrinology are now considered weighty matters of state. PMID- 17111924 TI - [Cytokines are a new generation of biotherapeutic agents]. AB - The review is dedicated to cytokines, a new prospective group of biotherapeutic agents. The author discusses the medical significance (real or potential) of interferons and other cytokines in treatment of viral infections as well as oncological and other diseases. PMID- 17111925 TI - [Present-day problems of occupation medicine]. AB - The author determines and substantiates the priority areas of scientific studies and presents the overall results of fundamental research into the topical problems of occupation medicine, such as the pathomorphosis of the modern forms of occupational and industrial diseases, new cell technologies and DNA-diagnostic technologies, occupational stress, the evaluation of health hazards, the combined effects of industrial factors, sociohygienic monitoring, reproductive health, harmonization, etc. as scientific basis for measures directed towards the improvement of health of working population in Russia. PMID- 17111926 TI - [On the mechanisms of thyrocyte proliferation and death in autoimmune thyroid diseases]. AB - Lymphocytes isolated from diffuse toxic goiter (Graves' disease, GD) stimulate the proliferation of "normal" thyrocytes (isolated from euthyroid goiter) in primary culture, and give them the properties of GD-thyrocytes (loss of sensitivity to the growth-promoting factors of FCS and lesser capacity of binding antibodies from GD patients' serum). The complement-free sera of GD patients (but not the sera of patients with Hashimoto's thyroiditis, HT) induce the death of "normal" thyrocytes more rarely than full-complement sera do. Both types of serum cytotoxicity are manifested on GD-thyrocytes much more rarely than on "normal" cells. The Fas-receptor on GD-thyrocytes in situ is expressed less than on "normal" and especially on HT-cells. The level of soluble Fas-ligand in the serum of some complement-free patients was found to be increased. These sera induce apoptosis in "normal" thyrocytes, but not in GD-cells nor in human skin fibroblasts. In the authors' opinion, the proliferation of GD-thyrocytes in situ is stimulated by intrathyroid lymphocytes, which directly stimulate this process and induce the loss of receptors which mediate the cytotoxic effects of serum factors. PMID- 17111927 TI - [Cholesterol is an important molecule in the processes of the synaptic plasticity and degeneration of neurons]. AB - The importance of homeostasis of neural tissue to neuron functioning, the synaptic plasticity of the hippocampus, and laboratory animals' behavior was demonstrated by the authors earlier. A range of experimental data evidences that cholinergic neurotransmission, ionotropic and metabotropic receptors, excessive tau phosphorylation, alterations in amyloid-beta biochemistry, oxidative reactions, and other features of neurodegenerative processes depend on the precise regulation of cerebral cholesterol metabolism. Such results suggest that disturbances in cholesterol homeostasis are the common primary cause of the sporadic and familial forms of Alzheimer's disease, Down syndrome, Niemann-Pick disease type C, and explain the similarity of neurodegenerative signs in different degenerative diseases of the nervous system. The present work was introduced at an annual conference of American Society for Neuroscience, and is available as a scientific report at www.neurobiologyoflipids.org/content/3/7/. PMID- 17111928 TI - [Contemporary views on the pathogenesis of atherosclerosis]. AB - Recent studies show that inflammation plays the key role inthe pathogenesis of atherosclerosis. Immune cells dominate at the initial stages of the atherosclerotic lesion of blood vessels. The effector molecules accelerate the progress of the lesion. This approach to the assessment of the pathogenesis of atherosclerosis makes it possible to search the ways for the prevention and suppression of immune inflammation. Nowadays vaccination against primary autoantigens is being successfully used for protection from experimental atherosclerosis. Modulation of immune response, involved in atherosclerosis, includes vaccination inducing immune protecting response of the development of tolerance by means of the process from Th1 to Th2 cell response. PMID- 17111929 TI - [The role of regulatory T-cells in autoimmune rheumatic diseases]. AB - Regulatory T-cells CD4+CD25+Foxp3, possessing suppressory activity, play the key role in the development of autoimmune diseases, maintenance of peripheral tolerance in transplantation immunity, and the prevention of a pathological immune response to intestinal microflora or microbial infection. A decrease in the total number of circulating CD4+CD25+ regulatory T-cells and their suppressive activity have been found in patients suffering from various autoimmune diseases, such as type 1 diabetes, multiple sclerosis, autoimmune hepatitis, psoriatic arthritis, juvenile idiopathic arthritis, and systemic lupus erythematosus (SLE). The authors of this study investigated the phenotypic characteristics of peripheral blood lymphocytes in 31 SLE patients and the effect of treatment on the content of CD4+CD25+ T-cells before and after pulse therapy with methylprednisolone and cyclophosphan. The total number of regulatory T-cells in the group of untreated patients was almost twice lower vs. the group of healthy donors. As a result of the therapy, the proportion of regulatory T-cells increased significantly, although it did not reach the values in the control group. The data from this research confirm the development of a defect of CD4+CD25+ T-cells at the active phase of SLE, and a possibility to partially correct this defect with an effective therapy. PMID- 17111930 TI - [Study of the influence of Staphylococcus aureus on gastrointestinal tract microbiocenosis in rats]. AB - The authors studied the modifying effect of Staphylococcus aureus on the microbial composition of gastrointestinal tract microbiocenosis. The subjects were female rats in the condition of eubiosis or dysbiosis. The species and quantitative composition of the fecal microflora and the parietal mucin in different parts of the intestine were studied after an intragastral administration of St. aureus suspension. A single introduction of St. aureus into the gastrointestinal tract of rats led to the appearance of this microbe in the feces and parietal mucin in all the parts of the intestine regardless the initial condition of the intestinal microbiocenosis. The indigenous microflora, both in eubiotic and dysbiotic conditions, practically did not respond to an intragastral administration of staphylococcus, except a little decrease in the proportion of bifidobacteria. Meanwhile, there was a significant increase in the incidence of candid detection. The indigenous parietal microflora changed more substantially, which demonstrates a higher sensitivity of the parietal microbiocenosis to a short-time exposure to an exogenous microbial factor. PMID- 17111931 TI - [Cell therapy as a method to correct pathogenetic disturbances in dyslipidemia and early atherosclerosis]. AB - The article is dedicated to the pathogenetic mechanisms of the development of immune inflammation in the vascular wall, as well as hepatic functional disregulation in dislipidemia and atherosclerosis. The authors emphasize that the decompensation of the process of cell regeneration in actively functioning organs (vascular endothelium, hepatocytes) and a concomitant immune disregulation become the main factors of the progression of dislipidemia and atherosclerosis. The authors consider the conditions for and factors of cell therapy, based on the use of allogenic fetal hepatic cells and autological bone marrow cells. PMID- 17111932 TI - [Viral infection and myocardial pathology]. AB - A viral infection is the most frequent cause of myocarditis. A viral invasion and viral-mediated cardiomyocyte lesion stimulate an immune system response directed towards the elimination of viral particles, the destruction of infected myocytes, and the limitation of viral dissemination. Numerous studies have shown that the balance of two subpopulations of CD4+ lymphocytes (T-helpers types 1 and 2) determines the character of the immune response in myocarditis, and that this balance is important at the initial stages of immunization, while the cytotoxic CD8+ lymphocytes participate in the myocardial lesion directly. Disorder in the elimination of activated lymphocytes' clones, the degree of which correlates with the degree of cardiac insufficiency, may be one of the mechanisms of myocarditis progression. Disbalance or defects in the system of the regulatory cytokine network also may lead to the activation of potentially autoreactive T lymphocytes. A connection between viral myocarditis and dilated cardiomyopathy (DCMP) has been discussed for many years. One of the reasons for DCMP development is a viral invasion into the myocardium. Proteinase A, which induces dystrophin proteolysis, which impedes its function, is an ingredient of enteroviruses. Hence, a viral infection may cause two myocardial diseases: viral (infective-and immune) myocarditis and DCMP. PMID- 17111933 TI - [Adiponectin, a new hormone: its role in the pathogenesis of diabetes mellitus]. AB - Adiponectin is a hormone of adipose tissue, activating lipid metabolism and other physiological functions. Adiponectin deficiency induces obesity and decreases insulin-regulated carbohydrate metabolism, thus leading to insulin resistance. Blood level of adiponectin falls in type 2 diabetes. Adiponectin receptors- AdipoR1 and AdipoR2--are proteins with 7 transmembrane domains, which are synthesized mostly in muscles and the liver and function in a close connection with G proteins. Obesity and diabetes lower the tissue concentration of the receptors, thus impeding adiponectin regulation of lipid exchange and lowering the effectiveness of the insulin control of carbohydrate exchange. Adiponectin also influences cardiovascular functions and prevents atherosclerosis and some of the other kinds of vascular pathology. PMID- 17111934 TI - [The pathogenesis, early diagnosis, and pharmacocorrection of fetal condition disturbances in high risk pregnancy]. AB - A longitudinal retrospective study of 803 women and a prospective study of 739 women were carried out. A complex system of the examination of women with a high placental insufficiency risk, starting from the first trimester, was developed; the appropriateness of the application of new technologies of placental system and fetal examination were substantiated. The authors offer optimal regimens of pharmacocorrection of fetal condition disturbances and the activation of compensatory and adaptive reactions of the fetoplacental complex in placental insufficiency of various origins. The optimal obstetric tactics, the possibilities of therapy and pregnancy prolongation, and indications to operative delivery were defined on the basis of the features of placental dysfunction pathogenesis and a thorough analysis of the features of placental insufficiency in women with a high risk of perinatal complications. PMID- 17111935 TI - [The role of dominant motivation in the formation of a goal-seeking personality]. AB - The article contains experimental data on the neurophysiological mechanisms of dominant motivations. Motivation is considered a component of the system architectonics of behavioral actions, closely connected with the mechanisms of reinforcement. The author shows that cortico-subcortical mechanisms constitute the energetic basis for dominant motivations. The apparatus of action result acceptor plays the role of the directing component of human goal-seeking activity. The author considers the role of action result acceptor in interaction with dominant motivation, in the molecular mechanisms of genetically determined and individually acquired memory, and in the forming of dynamic stereotypes. PMID- 17111936 TI - [The problems of the central regulation of immune system biorhythms: the role of exogenous and endogenous melatonin]. AB - One of the key synchronizers of mammalian circadian rhythm is the hormone melatonin (MT) produced by the pineal gland. MT is characterized by a wide spectrum of biological activity including its immunopotentiating effect. At the same time, the results of studies dedicated to the effects and mechanisms of melatonin immunoactivity seem to be contradictory. Studies conducted by the authors of this article show that diurnal variations in MT content in blood and saliva of healthy humans demonstrate significant individual fluctuations connected with the seasonal factor. The character of MT relations with the immune system varies depending on the phase of the circadian cycle. Patients with immunopathological condition (bronchial asthma--BA) demonstrate a reduced number of correlations between MT level and immunological parameters. After treatment, the positive dynamics of immunological and clinical parameters in BA patients is associated with an increased number of correlations between the immunological status parameters and MT level in blood, which may indicate an increase in its immunoregulatory role. Administration of melatonin and amitriptyline, the inductor of its synthesis, in animals kept under continuous light helped to recover normal circadian rhythmicity in the cell content of lymphoid organs. Solving problems associated with the immunomodulating and synchronizing role of MT could give researchers an insight into the details of neuroendocrine and immune interrelations and help to develop new methods for the prevention and correction of immune status disturbances in diseases and desynchronoses (stresses, jet lags, shift and night work, etc.). PMID- 17111938 TI - Now and then: a fountain pen, surgical scissors and a blue cardigan. AB - It was about 10 years ago that nurse education moved to the universities. Perhaps, this is a good time to consider what we think of this move. Inevitably, there were advantages and disadvantages to the 'old system' but there is more to it than that. Many of us have clear memories of the preliminary training school, living-in and the greater discipline in those days. These memories are often cherished and for good reason. To appreciate the full depth of change that has taken place we need to look a little further back than 1996, we need to look back to the 1950s and 1960s. This was the heyday of the school of nursing. PMID- 17111939 TI - Safe placement of nasogastric tubes in children. AB - Insertion of nasogastric tubes is a common nursing procedure but practice rituals and unfounded assumptions can compromise safety. Current methods for checking the position of nasogastric tubes are not always suitable in children but there is limited guidance on how to proceed when gastric placement cannot be determined. As a result of a local adverse incident, a working party challenged the assumptions and rituals of practice using an evidence-based framework. A nasogastric tube algorithm was developed using a risk assessment approach to improve the safety of children in hospital and community settings. PMID- 17111940 TI - Infection control: 'now wash your hands'. PMID- 17111941 TI - Giving children and parents a voice--the parents' perspective. AB - Being the parent of a sick child requires advocacy, endurance and persistence if the child's voice is to be heard and his or her needs met. Based on her experiences as the mother of a child who was treated for leukaemia and died at the age of nine and a half years, Bridget Taylor illustrates the difference it makes when professionals make time to listen and take on the advocacy role for the parents. She argues that there is much more that nurses can do to know the child, to understand the parents' distress and then to create a culture that gives permission for parents and children to disagree with or question the care they receive. PMID- 17111942 TI - Improving hospital experiences for children with complex needs. PMID- 17111943 TI - Learning to communicate with children with disabilities. AB - A positive incident during a placement in a special school is used to illustrate that reflecting on positive incidents helps the student nurse to recognise good practice and personal strengths. The incident involving a child's achievement in mastering new communication skills led to improved understanding of the role of the special educational needs teacher and the speech and language therapist and the positive effect of early professional intervention. It also emphasised the benefits of Makaton and picture aids for children with communication problems. A major benefit of the placement experience was that it improved the student's confidence in approaching and interacting with children with special needs and speech difficulties. It also highlighted the value of nurses learning basic Makaton in order to enhance their communication skills. Using reflection to review positive experiences can be as effective in highlighting strengths and weaknesses as reflecting upon negative experiences. PMID- 17111944 TI - Improving care for children requiring surgery and their families. AB - Despite the publication over many years of guidance and policies related to children in hospital and children having surgery, concerns remain about the standards of care provided to children and families. A survey of 63 nurses working with children was carried out by members of the Royal College of Nursing Children's Surgical Nurses Forum to evaluate the extent of implementation of guidance related to: children's choices in relation to theatre garments, whether parents or the main carer can accompany the child into the anaesthetic room and be with their child in the recovery area, and which healthcare professionals are available to support the child and family during transfer to theatre and in the theatre environment. Forty four nurses responded (70 per cent). Results indicated that at least one parent was almost always given the opportunity to be present at induction and in recovery. Only 36 per cent said that children are always offered a choice of what to wear The presence of qualified children's nurses in theatres was reported to be limited. Policy principles and guidance documents can be used by nurses to influence new ways of working to ensure practices are appropriate for children undergoing surgery. PMID- 17111945 TI - An inter-professional approach to children's nursing education. AB - Nurses caring for children and young people do not practice in isolation but work as members of a multi-professional healthcare team. The Inter-professional programme delivered at Canterbury Christ Church University offers pathways in midwifery, adult nursing, child nursing, medical imaging, occupational therapy, social work and operating department practice. Benefits include the focus on children and their families from the outset, and learning to nurse using a multi professional approach. Challenges in implementing the programme include maintaining professional identity, changing patterns of teaching and learning, supporting clinical staff in adapting to early student placement and the multiprofessional focus. PMID- 17111946 TI - Observing the sick child: Part 2a. Respiratory assessment. AB - As the focus on evidence-based healthcare delivery gains momentum, nurses need to critically reflect on their traditional practice base. Assessment is the cornerstone of safe and effective practice and goes hand in hand with the process of prioritising a sick child's needs. This article begins with a practical and critical review of the validity and reliability of basic respiratory assessment focused on measurement of respiratory rate, rhythm and depth. Part 2b in February 2007 will be a practical step-by-step introduction to the theory and practice of advanced respiratory assessment using palpation. Part 2c in the following month will focus on respiratory auscultation methods. PMID- 17111947 TI - Pay cut after 'best year'? PMID- 17111948 TI - Battle lines drawn as unions reject Brown's squeeze on pay. PMID- 17111949 TI - Lost in translation. AB - Although there are large numbers of people in the UK who are hearing-impaired or cannot speak English, signing and translation services in the NHS are all too often woefully inadequate. PMID- 17111950 TI - Standing up for dignity. AB - Senior nurses say that older people's dignity is an issue for all health staff. While further guidance from the Department of Health is anticipated, some nurses are already innovators in this area. PMID- 17111951 TI - A personal lesson in Alzheimer's care. PMID- 17111952 TI - Socialisation of seconded healthcare assistants. AB - AIM: To compare the clinical practice experiences of seconded healthcare assistants (HCAs) with four major socialisation concepts. METHOD: The sample was comprised of eight seconded HCAs, all from the same pre-registration mental health nursing cohort. A semi-structured focus group strategy was used, with the group convening on a yearly basis. FINDINGS: The practice learning experiences of seconded HCAs are different to other nursing students and could be viewed as a unique socialisation process. CONCLUSION: Course planning teams should consider socialisation factors and supportive mentor strategies when designing the first year of pre-registration nursing programmes. PMID- 17111953 TI - Assessment and treatment of older patients with constipation. AB - This article examines the experiences of an older patient with constipation to illustrate the importance of thorough assessment. The interaction between another medical condition and constipation is discussed, and indications for laxative use are outlined. The current evidence base for bowel care is limited. Tools for risk assessment are required and further research is needed to improve patient care in this area. PMID- 17111954 TI - Principles of aseptic technique in urinary catheterisation. AB - Most nurses are aware of the importance of aseptic technique but some may be unsure about applying the technique during urinary catheterisation. This article explains the principles of aseptic techniquue and their application to the procedure of urinary catheterisation. PMID- 17111955 TI - Identifying sepsis. PMID- 17111956 TI - Shifting priorities. PMID- 17111957 TI - Alternative nursing. PMID- 17111959 TI - Baptism of fire. PMID- 17111958 TI - Spotlight on success: Nancy Fontaine. PMID- 17111960 TI - Involving patients in research. PMID- 17111961 TI - Great expectations. PMID- 17111962 TI - Formula for success. PMID- 17111963 TI - Nursing and information technology. PMID- 17111964 TI - A development programme for senior nurses. PMID- 17111965 TI - Crossing boundaries: a staff exchange initiative. PMID- 17111966 TI - Promoting public health: Part 1. PMID- 17111967 TI - [Pathological fracture revealing an osseous histoplasmosis. A case report on a 60 year patient]. AB - The authors report a new case of African Histoplasmosis in a 60-year-old patient. It was an humeral localization revealing a pathological fracture which grew into an extension of osteolysis and a cutaneous fistulization likely to be a malignant bone tumor. The case has been diagnosed by surgical biopsy and histological analysis. Its antifungal treatment in progress resulted in the drainage of the out-flow that should permit the bone reconstruction by graft. The authors stress on the need to focus on this affection whenever, in a tropical area, one is faced with any chronic bone fistula that cannot positively be cured in spite of sound medical cares. PMID- 17111968 TI - [Relapses after surgical treatment of Buruli ulcer in Africa]. AB - Very few documents are available on Buruli ulcer (BU) in the world in general and in Africa in particular, even though this continent is the worst hit by Buruli ulcer. Information has been reported by hospitals protocols. The BU treatment in Africa is mainly based on surgery The results of these treatments vary from one study to another as the relapses rates show it: 17% in Kanga et al. in Cote d'Ivoire, 16% in Amofah et al. in Ghana, 6% in Debacker et al. in Benin. A manual on management of Mycobacterium ulcerans has been written up by the World Health Organisation (WHO) in 2001, mainly focusing on surgery management. In 2004, WHO also suggested the use of antibiotics associated or not according to cases with surgery Results are evaluated in terms of recovery, relapses or recurrences. This review of the literature on BU treatment by surgery in Africa from 1920 to 2005 draws up the frequency of relapses after surgery in BU while pointing up the usefulness of a standardization of the evaluation indicators of the real efficiency of BU surgical treatment. PMID- 17111969 TI - [Delayed autologous transfusion: about 70 surgical patients at the Saint-Jean-de Dieu Hospital in Afagnan, Togo]. AB - This prospective study was conducted from 2000, September the 1st to October the 31st, at the Saint-Jean-de-Dieu Hospital in Afagnan (Togo). All the 75 patients who received indication of autologous blood transfusion (ABT) accepted the protocol; 70 out of them, among whom 63% were female, benefited an ABT The age of the patients varies between 13 and 80 years old (average 33.2). Initial rate of haemoglobin was on average of 11.7 g/dl; 14% of these patients had a haemoglobin diseases. The most frequent interventions were hysterectomies (21%), bone surgery (16%), prostatectomies (11). Only one blood unit was taken from 63 patients and two units from the 7 others inducing giddiness (5) and headaches (1). During intra and postoperative periods, 41 patients were given 45 units of blood (use rate: 58%). The percentage of patients who received transfusion was 56% in gynaecological surgery 79% in orthopaedics and 88% in urology. Only one patient received an additional homologous unit of blood. The rate of haemoglobin was on average 10.2 g/dl the day after surgery. No incident in connection with the ABT was recorded. The ABT is a feasible, effective and secure method in the context of a small African hospital. It may be a solution to the problems of shortage of blood products and transfusion safety. Training and motivation are necessary for its successful implementation. PMID- 17111970 TI - [Influence of fetal haemoglobin rate (FHb) on the oxidizing stress in homozygote sickle cell patient living in Abidjan, Cote-d'Ivoire]. AB - Sickle cell anemia being involved in oxidizing stress, our objective was to study the influence of the fetal haemoglobin rate (FHb) on the lipoperoxidation markers in homozygote sickle cell patient in tropical African surroundings. The study population was composed of 73 subjects among whom 57 homozygote sickle cell subjects and 16 healthy control cases. These subjects were distributed in 4 groups according to FHb rate: group 1 (FHb rate under 10%), group 2 (FHb rate ranging from 10 and 20%), group 3 (FHb rate above 20%), group 4 (control cases with no sickle cell disease). On the biological level, the markers of plasma lipoperoxidation represented by substances reacting with thiobarbituric acid (TBARS) significantly increased in sickle cell patients comparatively to control cases (p = 10(-6)). A strong positive correlation (r = +0,70, p<0,01) was found between HbS and the TBARS rate. Comparison of biological parameters of homozygote sickle cell patients according to HbF rate shows that TBARS rate is all the more low as the HbF rate is high (p = 0,02). Moreover the number of irreversible and reversible sickle cells is higher in the group 1 which has the highest rate of TBARS. This observation is confirmed by a coefficient of positive correlation between TBARS and reversible sickle cells (r = +0,40, p < 0,01). This study strengthens the role played by HbF on the modulation of physiopathology of homozygote sickle cell anemia by the control, among others, of free radicals. PMID- 17111971 TI - [Management of rhesus alloimmunisation by spectrophometry: about one case at the Yopougon Teaching Hospital, Cote-d'Ivoire]. AB - The authors report a care management of a patient presenting a severe alloimmunisation treated by spectrophotometry, a very efficient method but difficult of access in developing countries. Beyond the restrictive paraclinical exams which do not allow an efficient monitoring of this pathology, the lack of alloimmunisation prevention during illegal abortions and the lack of information about patients'medical files are highly responsible for the difficult management of Rhesus- negative patients. However the authors remind that only the scan at best combined with the Doppler, are effective exams helping to avoid the constraints of spectrophotometry. Although quite well-known this pathology induces very severe foetal consequences and may involve the obstetric prognosis of young women. The caesarian section will still remain for a long time the only means to prevent foetus from alloimmunisation complications in developing countries. PMID- 17111972 TI - [Acceptability of HIV screening in patients suffering from sexually transmitted infections at the antiveneral center of Abidjan, Cote-d'Ivoire]. AB - The systematic screening in high risk groups can be an efficient way to struggle against HIV/AIDS. We have conducted a transversal descriptive study in patients suffering from STI and who had consulted the antiveneral dispensary of the Institut National de Sante Publique (Abidjan) from 1st September 2002 to 31st October 2002. The aim was to identify the acceptability of the HIV screening test. During this inquiry, 167 patients were recruited. This study permits to confirm the young age of the population with a male predominance in patients having sexual transmitted infections. So, 70.1% have accepted to do this test. However, 23.4% of patients recruited had really done the HIV test. Among those who have accepted to do the test, 28.2% were infected. PMID- 17111973 TI - [Intestinal parasitosis in French West Indies: endemic evolution from 1991 to 2003 in the University Hospital of Pointe-a-Pitre, Guadeloupe]. AB - Guadeloupe is one of the French West Indies, where, until 1960, intestinal parasitic infections were endemic. In the microbiological laboratory of the university hospital of the island, we carried out a retrospective study of the 17,660 stool examinations received from 1991 to 2003. All the stool specimens were examined using at least a wet mount preparation and a concentration method (Bailenger). Specific techniques such as Ziehl-Neelsen modified acid fast stain, chromotrope staining procedure or Baermann's technique were used when indicated. A parasite considered as really pathogenic is detected in only 5.6% of the cases of the 17,660 stool examinations. The intestinal protozoa count for 10.8% of them, G. intestinalis (60%) is the most common, followed by Cryptosporidium sp (26%) and Isospora belli in AIDS patients while Entamoeba histolytica/dispar rarely appears (2.3%). Among the parasitic intestinal helminths, Strongyloides stercoralis is seen as the most prevalent (82%) whereas hookworms and Trichuris trichuria count respectively for 9.8% and 5.6%. Schistosoma mansoni appears as an exception. This epidemiological change for intestinal parasitic infections results from the improvement of socio-economic and sanitary way of living of the population. However the persistence of the strongyloidiasis constitutes the outstanding fact in a area of strong prevalence of the HTLV1 infection. PMID- 17111974 TI - [Schooling of the child and teenager suffering from sickle-cell disease in 2004, Brazzaville, Congo]. AB - A case-control transversal study has been carried out in March 2004 in the paediatric unit of the Brazzaville teaching Hospital to assess the impact of the sickle cell disease on school-age children. School-age children and teenagers of both sex were distributed in 228 homozygote sickle cell children (group I) aged of 5 years old and 8 months old to 21 and 245 children recruited in state schools aged of 5 years old and 6 months to 19 years old. In the group I, 78 children suffering from sickle cell disease are ahead in their school achievement, 59 have a normal education and 91 meet some difficulties with no significant difference; in the group II, 122 children are ahead in their school achievement, 81 have a normal education and 42 meet difficulties (p < 0,001). The average of children having school difficulties was 2,4 +/- 1,5 years in the group I with extremes ranging from 1 to 3 years. By comparing both groups in primary school, no difference was to be found in children doing well at school and children having a normal education: 54 children of the group II are reported being ahead in their school achievement (39, 7%) and 81 children in the group II (55, 1%); normal education for 38 children of the group 1 (28%) and 58 children in the group II (39, 5%). On the other hand, 44 children are having school difficulties in the group I (32, 3%) against 8 children in the group II (5, 4%) (p < 0,001). As a matter of form we have observed that ahead schooling is to be found in 24 sickle cell children at HbF < 10% (41, 4%) and 54 sickle cell children at HbF < 10% (34, 2%) (p < 0,05); normal education in 14 children with sickle cell disease at HbF > 10% (24, 1%) and 45 children with sickle cell disease at HbF < 10% (25, 9%) and school difficulties in 20 children with sickle cell disease at HbF > 10% (34, 5%) and 71 children with sickle cell disease at HbF < 10% (39, 9%). PMID- 17111975 TI - [Bacterial meningiditis in adult patients in Central African hospitals]. AB - Bacterial meningitidis of the adult remain frequent and serious affections in spite of therapeutic progress. The authors report results from a retrospective study of 502 cases over 5 years. Hospital prevalence was 12.1%. These infections occur during the dry season between November and April in 75% of the cases. An important peak was observed in March, that is to say 24.5%. Patients' average age was 34.7 years old with extremes ranging from 15 to 80. The isolated micro organisms were: the pneumococcus (45.2%), the meningococcus (14.5%), salmonella (1.6%) and the Haemophilus influenzae (1.2%). Meningitidis decapitated by a preliminary antibiotherapy represented 37,5%. Seroprevalence of the HIV infection during this work was 55.1%. Patients suffering from pneumococcus meningitis were contaminated by the HIV in 51.7% of the cases. Those suffering from meningococcus infection were infected with HIV in 52.9% of the cases. Those presenting meningitidis to Haemophilus influenzae and salmonellas had contracted the HIV respectively in 83% and 87.5% of the cases. The average duration of hospitalization was 14.2% days. The average mortality rate was 31.7% for the pneumococcus, 30. 1% for the meningococcus one. 53.4% of the patients recovered. PMID- 17111976 TI - [Air pollution in urban area in Cotonou and Lokossa, Benin]. AB - In some African countries, time of clean environment with no air pollution belongs to the past. From now on, pollution is a problem of public heath in addition to malnutrition and infectious diseases still badly controlled. The sanitary consequences induced by pollution is now a cause for concern in the population. Yet, very few data are available to develop awareness of the population and convince the authorities to implement urgent policy in order to put a brake on the phenomenon. The objective of this document aims at describing the atmospheric pollution recorded in Cotonou and Lokossa as well as possible symptoms linked to exposure of pollutants of the population. Therefore we carried out a transversal study in the city of Cotonou and Lokossa (125 kilometers from Cotonou). Air samples were taken at crossroads levels and motorbikes to measure the different chemical components, 400 taxi drivers were questioned on symptoms affecting them. The toxicological analysis of air and blood sample is carried out at the laboratory of the Universite Libre in Brussels. The data analysis revealed a high pollution in Cotonou. The carbon monoxide concentration varies from 26 ppm to 38,6 ppm in the morning and increases in the afternoon (58 to 78,6 ppm). Benzene is found with an average of 7,2 micro/m3 in Cotonou. These concentrations are not so high in the town of Lokossa (carbon monoxide from 45 to 51ppm). In Cotonou motorbike taxis account for 90% of the cases with intoxication symptoms 1,5 time higher than in the non-drivers of these motorbikes. The symptoms recorded on statement are intoxication disorders such as conjunctival hyperemia (18%) among which 12% of lacrimation, respiratory disorders (23%). On the other hand, registered disorders were not so important in Lokossa: conjunctival hyperemia (5,6%), nausea (32%) and other signs as sight disorders (4%). The disorders frequency in non drivers is not so high in Cotonou and Lokossa. In the blood test, the Hbco rate is abnormal in more than 66% of the drivers. The authors suggest the regular checking of imported secondhand cars, measures to reduce exhaust emissions and the reinforcement of the law on air pollution as well as a proper management of crossroads. PMID- 17111977 TI - [Onchocerciasis in the area of Lastourville, Gabon. Clinical and entomological aspects]. AB - The African Programme for Onchocerciasis Control (APOC) has implemented a series of surveys aimed at evaluating the long-term impact of its activities. The region of Lastourville (Gabon) is one of the selected sites for this study. A total of 886 persons was examined for skin lesions, and 459 out of them participated in detailed ocular examinations. Blackflies were collected during one year and dissected. Although the focus was found to be hypoendemic (prevalence of nodules: 7.7%), the frequency of onchodermatitis was relatively high. The lesions of the anterior segment of the eye were rare, but the prevalence of optic nerve disease, and of choroido-retinal lesions reached 5.2 and 2.7%, respectively. The annual transmission potential (2,171 infective larvae per man) was high, when compared with the results recorded in the human population. This may be due to the presence of Onchocerca spp. of animal origin in the blackflies. These results indicate that in the area of Lastourville, though regarded as the main focus of onchocerciasis in Gabon, the disease is relatively mild. PMID- 17111978 TI - [African Programme for Onchocerdasis Control (APOC): Onchocerca Simulium squamosum in two regions in the Republic of Cameroon]. AB - In 1998, the African Programme for Onchocerciasis Control (APOC) decided to launch a long-term impact assessment of its operations. This paper reports the baseline entomological data collected throughout a whole year in two sites of Cameroon (Kahn and Bolo). The Simulium populations of the two study sites were characterized by parous rates of 7.2% and 33.5% respectively and infectivity by O. volvulus of 31 and 190 infective larvae per 1000 parous flies respectively The Annual Transmission Potentials (ATP) were respectively 523 and 9972 infective larvae per man and per year in Kahn and in Bolo. The Simulium populations studied in both sites, even though the ATP in Kahn is 19 times lower than that of Bolo, showed a pattern of an onchocerciasis hyperendemic zone in terms of vector capacities and entomological indices. PMID- 17111979 TI - [Pyrethroid and DDT resistance of Anopheles gambiae s.s. (Diptera: Culicidae) in five agricultural ecosystems from Cote-d'Ivoire]. AB - The promotion of pyrethroid impregnated bednets among the populations is a major activity of the National Malaria Vector Control Programs in African tropical countries. However pyrethroid resistance in Anopheles gambiae, a major malaria vector, has been observed in several parts of Cote-d'Ivoire since 1993. As insecticides used in agriculture against pests are frequently considered as important factors responsible for resistance in malaria vectors, we have evaluated insecticide resistance of An. gambiae populations taking into account the main crops cultivated in Cote-d'Ivoire. Five areas were selected according to the use of insecticide either in agriculture or for domestic purposes: a urban area where vegetables and rice are main crops, a rural rice growing area, a rural area of coffee/cocoa production, a rural area of fruit farming and a rural area without any use of agricultural insecticide. Susceptibility tests were carried out using WHO diagnostic test kits. About 6500 females of An. gambiae were exposed to insecticide impregnated papers (permethrin 1%, deltamethrin 0.05%, DDT 4%) for one hour. Results confirmed that both mortality rates and knockdown time analysis were important to study the resistance data. By using mortality rates, populations of An. gambiae were found to be resistant to pyrethroids and DDT in four of the five areas. Resistance was the highest in urban area, lower in fruit and coffee/cocoa areas and at low level in rice growing area. An. gambiae from area without agricultural treatment was found susceptible to pyrethroids but slightly resistant to DDT A significant increase of knockdown time was observed in all areas with the 3 insecticides. These results agreed with previous studies showing that kdr mutation was the main resistance mechanism to pyrethroids in An. gambiae populations in Cote-d'Ivoire. They also agreed with knockdown time which is an early indicator of resistance development for the population in area without agricultural treatment. In this population the frequency of homozygous resistant individuals was probably too low to have a significant decrease of mortality rates to pyrethroids. PMID- 17111980 TI - [Feeding pattern of Rift Valley Fever virus vectors in Senegal. Implications in the disease epidemiology]. AB - During the rainy season 2003, an entomological survey was undertaken in the Sahelian bioclimatic zone of the Ferlo area in northern Senegal, in order to evaluate the degree of interaction between Rift valley fever (RVF) virus vectors and domestic animals and to determine the role of natural vertebrate hosts in the transmission and maintenance cycle. The study of vector-host contact was carried out under bed net traps using man, cow, sheep, chicken as bait whereas the RVFV vectors-vertebrate host interactions were studied through the analysis by an ELISA technique of the origin of the blood meals from naturally engorged females collected by aspiration. Blood meals sources were determined using a set of eight antibodies. Overall, the different known RVFV vectors (Culex poicilipes, Aedes vexans and Aedes ochraceus) were opportunistic although the bovine-baited net was, as far the more effective trap with 53.6% of collected mosquitoes. It was followed by the sheep-baited net (16.7%), man-baited net (12.6%) and chicken baited net (11.6%). The more effectiveness of the bovine-baited net confirms the degree of implication of this host in RVF epidemiology. The study of vector-hosts interactions in nature showed that among the 1,112 mosquito blood meals tested, 701 were identified of which 693 were from Aedes vexans. The percentage of non reacting blood meal was 36.7% whereas 16.9 % of the blood meals were taken at least on two vertebrate hosts. Overall, 53.2% of the blood meals from Ae. vexans were taken on equine, 18.6% on bovines, 7.1% on sheep and 0.6% on human. No blood meal was taken on rodent. The greatest diversity was observed in August. These host feedings patterns show that although equine is known to play a minor role in RVF epidemiology a thorough attention should be made to this host with regard to the percentage of blood meals taken in this host. The low percentage of blood meals taken on human could probably explain the low human infection rate observed up to now in Senegal. With the high percentage of non-reacting blood meals, our results also underline the necessity of an improvement of ELISA techniques and the use of more reliable tools as molecular markers for an exhaustive identification of vertebrates hosts involved in RVF epidemiological cycle. PMID- 17111981 TI - [Malaria: knowledge, behaviour and practices among a rural population of Gossas, Senegal]. AB - Malaria remains a major public health problem in Sub-Saharian Africa, in terms of morbidity and mortality rate. To assess the knowledge and behaviour of population regarding the transmission, the treatment and the prevention of malaria, we conducted a cluster sample household survey in Gossas, a rural District in Senegal, from May 2nd to May 6th 2005. A questionnaire that focused on socioeconomic conditions, beliefs, knowledge about and behavior toward antimalarial medication and the prevention means used was given to 480 household owners. Overall, 107 pregnant women and 1,201 children aged less than 5 years old lived within these household. More than a half of the household owners (51%) were illiterate and 25.2% ignored how malaria is transmitted. Fever was the most common symptom suggesting malaria (61%). In 46.1% of febrile cases, people did not seek for treatment from a physician. Home treatment of febrile episodes was based on paracetamol or aspirin (84%), chloroquine (13%) and cotrimoxazole (2.9%). Overall, the proportion of insecticide treated nets users were 22.7%. This percentage was 14.9% and 11.4% for pregnant women and children younger than 5 years old, respectively. People having radio sets, regular access to television, and people aware of the transmission route of malaria were more likely to use bed nets. In most cases, organic material burning was used as repellent against mosquitoes. The low prevalence of bed net use was most often explained by participants' limited accessibility to and by the high cost of insecticide-treated nets. Knowledge about malaria prevention and treatment is low in the rural district of Gossas. The rate of insecticide-treated-bed nets use in vulnerable people is very low, far from the Abuja meeting objective. A sensibilization program and a social marketing plan for insecticide-treated-bed nets could improve this situation. PMID- 17111982 TI - [New imaging techniques, new demands. Precised problems and subspecialty competency needed for good results]. PMID- 17111983 TI - [Lack of a unified, national practice for compulsory care of children and adolescents. A questionnaire reveals notable regional variations]. PMID- 17111984 TI - [Treatment of patients with skin and soft tissue infections. Results from the STRAMA survey of diagnoses and prescriptions among general practitioners]. PMID- 17111985 TI - [PET/DT can cause clinicians extra concern . A case report illustrates the dilemma of judging additional findings]. PMID- 17111986 TI - [Mindfulness meditation--an old fashion method for stress relief]. PMID- 17111987 TI - [Motivational interviewing--better than counseling]. PMID- 17111988 TI - [A lot to gain with CBT in pain rehabilitation]. PMID- 17111989 TI - [Promising therapy of borderline personality disorder]. PMID- 17111990 TI - [Status of the evidence: psychotherapy is efficient, but many questions still unanswered]. PMID- 17111991 TI - [Psychotherapeutic skills important in primary health care]. PMID- 17111992 TI - [Impaired absorption of antibiotics in diarrhea?]. PMID- 17111993 TI - [Systemic effect of nasal corticosteroids?]. PMID- 17111994 TI - [Time for possibility of compulsory treatment in psychiatric outpatient care]. PMID- 17111995 TI - [Dear colleagues, we have missed something!]. PMID- 17111996 TI - [Modernize the CBT concept in Sweden!]. PMID- 17111997 TI - [Final comment: free abortion a necessity]. PMID- 17111998 TI - Cancer pain management in the 21st century. AB - Cancer causes pain as it invades bone, compresses nerves, produces obstructive symptoms in the pulmonary, gastrointestinal, and genitourinary systems, and distends involved visceral organs. This manuscript reviews progress in cancer pain management during the past 2 decades. Since the 1980s, we have seen (1) genuine advances in research on the biology of pain, (2) new approaches to the treatment of cancer pain, and (3) important changes in the health-care system to ensure that pain is appropriately assessed and managed. Currently, clinicians have the appropriate diagnostic and therapeutic tools to ensure that the vast majority of patients with cancer pain can be comfortable during their illness. Nevertheless, too many patients with terminal malignancies continue to die in pain in nations around the globe. An effective strategy to make alleviating pain a major health-care priority remains the primary challenge to effectively palliating patients with cancer pain. PMID- 17111999 TI - Treating advanced breast cancer in the older woman. AB - As half of all breast cancers occur in patients beyond the age of 65 and a quarter beyond the age of 75, a significant number of patients with metastatic breast cancer are elderly. New hormonal therapies, such as aromatase inhibitors, appear to have favorably improved the survival of these patients. Side effects such as osteoporosis or cognitive issues appear manageable. Information specific to elderly patients has recently emerged in the field of chemotherapy for metastatic breast cancer. This article reviews data on anthracyclines, taxanes, capecitabine (Xeloda), gemcitabine (Gemzar), trastuzumab (Herceptin), and bevacizumab (Avastin). For most patients in this setting, sequential single-agent chemotherapy appears at this time to be the preferred course of treatment. PMID- 17112000 TI - Cetuximab-associated infusion reactions: pathology and management. AB - Cetuximab (Erbitux), a chimeric antiepidermal growth factor receptor monoclonal antibody currently used to treat metastatic colorectal cancer, is in clinical development for several other solid tumors. Although cutaneous manifestations are the most common toxicities associated with cetuximab, they are rarely life threatening. Cetuximab-related infusion reactions are less common, but they may become severe and cause fatal outcomes if not managed appropriately. Little about the specific etiology of these events is known; however, an overview of infusion reactions observed with other compounds may shed some light and help characterize cetuximab-related reactions. For physicians administering cetuximab, familiarity with acute reaction treatment protocols and preparedness to identify and manage symptoms promptly and effectively are most important to minimize potential risks. PMID- 17112001 TI - Topotecan in combination with cisplatin for the treatment of stage IVB, recurrent, or persistent cervical cancer. AB - PURPOSE: Topotecan, a camptothecin analog previously approved for the treatment of ovarian cancer and small-cell lung cancer, was granted regular approval by the US Food and Drug Administration (FDA) on June 14, 2006, for use in combination with cisplatin to treat women with stage IVB, recurrent, or persistent carcinoma of the cervix not amenable to curative treatment with surgery and/or radiation therapy. The purpose of this summary is to review the database supporting this approval. EXPERIMENTAL DESIGN: In a randomized multicenter study enrolling 293 eligible patients, topotecan plus cisplatin (TC) was compared with cisplatin monotherapy. The TC regimen consisted of cisplatin 50 mg/m2 IV over 1 hour on day 1 and topotecan 0.75 mg/m2 IV over 30 minutes on days 1, 2, and 3 every 21 days. RESULTS: There was a clinically relevant and statistically significant improvement in overall survival in the TC treatment arm. Median overall survival was 9.4 months (95% confidence interval [CI]:7.9-11.9) in the TC arm, compared to 6.5 months (95% CI:5.8-8.8) with cisplatin alone. The unadjusted hazard ratio for overall survival between treatment arms was 0.76 (95% CI: 0.59-0.98, P = .033) favoring the combination arm. The most common toxicities with TC included myelosuppression, nausea and vomiting, mucositis, rash, and hepatotoxicity. CONCLUSIONS: This report describes the FDA's review supporting this first approval of a chemotherapeutic drug for advanced cervical cancer based on demonstration of a survival benefit. PMID- 17112002 TI - Endoscopic ultrasound fine-needle aspiration in the staging of non-small-cell lung cancer. AB - Precise mediastinal staging of non-small-cell lung cancer is extremely important, as mediastinal lymph node metastases generally indicate unresectable disease. Reliance on computed tomography (CT) and positron-emission tomography (PET) alone to stage and determine resectability is limited by false-positive results. Whenever possible, pathologic confirmation of metastases is desirable. Mediastinoscopy and transbronchial fine-needle aspiration are widely established but imperfect modalities. Endoscopic ultrasound fine-needle aspiration (EUS-FNA) has emerged as a diagnostic and staging tool because of its safety, accuracy, and patient convenience. We reviewed 13 prospective studies evaluating the comparative performance of EUS for staging lung cancer. We conclude that EUS is a valuable staging modality. Further studies of the role of EUS compared to other modalities such as integrated PET/CT and endobronchial ultrasound (EBUS) are forthcoming. PMID- 17112003 TI - Woman with a papillary pulmonary lesion and a complex renal cyst. PMID- 17112004 TI - Current approach to pheochromocytoma. PMID- 17112005 TI - No scientific challenge was too daunting. PMID- 17112006 TI - Investment in midwifery research. PMID- 17112007 TI - Breastfeeding with HIV. PMID- 17112008 TI - Conference season--opinions voiced. PMID- 17112009 TI - The NHS pension scheme review--future members. PMID- 17112010 TI - Mentoring in midwifery. PMID- 17112011 TI - Homeopathy for childbirth: remedies and research. PMID- 17112012 TI - Modernisation: how the UMF grant was spent. PMID- 17112013 TI - Making research count. PMID- 17112014 TI - The future starts here! PMID- 17112015 TI - Antenatal care in children's centres--making it happen. PMID- 17112016 TI - Giving out the wrong signals. PMID- 17112017 TI - Will the midwifery profession develop if midwives remain politically naive? PMID- 17112018 TI - [Using instantaneous spectra to determine dominant species in the DDT process of epoxypropane]. AB - After solving problems of weak light detection, the calibration of the spectral sensitivity of the measuring system, and the synchronization of the measuring system, instantaneous emission spectra of epoxypropane in the process of deflagration to detonation transition (DDT) with the exposure time of 2-8 micros and the resolution of 0. 2 nm were acquired from six different side windows of an explosion shock tube. Using the corrected spectral data, curves of the optical radiant intensity of main reaction products versus the DDT distance from the ignition point were obtained. These curves provided information about the evolution of the reaction and the products during the DDT process. Results indicate that the chemical reaction rate of the gaseous fuel and the corresponding concentrations of intermediate products increased gradually at the deflagration stage, but at the moment of deflagration to detonation transition, the reaction rate increased rapidly and the concentrations! of products increased sharply. Among these main products, concentration increments of molecule CO, and radicals CHO and OH were greater than other products, which means that CO, CHO and OH are the dominant species that affect the DDT process greatly. PMID- 17112019 TI - [Experimental study of soft X-ray radiation in the interaction of circularly polarized femtosecond-laser-pulse with low pressure xenon]. AB - Using flat-field grating Spectrometer, the ions lines with wavelength between 5 and 60 nm were measured, which were produced by the interaction of circularly polarized 35 femtosecond ultraintense and ultrashort laser-pulse with 5 mm length xenon at the pressure 2 and 3 kPa respectively. The highest transition is the XeVIII: 4d10 5s(2 S1/2)--4d9 5s5p('P3/2) line at wavelength 17.0856 nm at 2 kPa and 3 kPa, the highest transition is 11.343 nm line of XeVII 4d10 5s2(1S0)--4d9 5s5f(3P1) transition. The xenon is ionized to XeVII, XeVIII and XeIX at both pressure. PMID- 17112020 TI - [Energy-pooling collisions of rubidium atoms: Rb (5P(J)) + Rb (5P(J))--> Rb (5S) + Rb (nl = 5D,7S)]. AB - An experimental study of rubidium energy pooling collisions, Rb(5P(J)) + Rb(5P(J))-->Rb(nlJ') + Rb(5S), at thermal energies, was carried out in a cell. Atoms were excited to either the 5P 1/2 or 5P 3/2 state using a single-mode diode laser. The excited atom density and spatial distribution were mapped by monitoring the absorption of a counter-propagating single-mode diode laser beam, tuned to either 5P 1/2-->5D 3/2 or 5P 3/2-->7S 1/2 transition, which could be translated parallel to the pump beam. The excited atom densities were combined with the measured fluorescence ratios to determine cross sections for the rubidium energy pooling process. For 5P 3/2 excitation the cross sections for nlJ' being 5D 5/2, 5D 3/2, and 7S 1/2 are (1.32+/-0.59) x 10(-14), (1.18+/-0.53) x 10(-14) and (3. 21+/-1. 44) x 10(-15) cm2, respectively. For 5P 1/2 excitation the cross sections for nlJ' being 5D 5/2 and 5D 3/2 are (6.57+/-2.96) x 10(-15), and (5.90+/-2.66) x 10(-15) cm2, respectively. The results were compared with those of other experiments. PMID- 17112021 TI - [Photoelectron decay properties of doped AgCl microcrystals under chemical sensitization]. AB - The photoelectron decay characteristic directly reflects the photographic efficiency of silver halide crystals. Measurement of the electronic decay time resolved spectrum of silver halide microcrystals can provide important information about the photoelectron decay action in latent image formation process. In order to know the influence of shallow electron trap dopant K4 Fe (CN)6 and S+Au on photoelectron decay, the photoelectron decay time-resolved spectra of AgCl emulsion doped by K4 Fe(CN) and that doped by K4 Fe(CN) firstly and then sensitized by S+Au were detected by microwave absorption dielectric technique, which can be used to study the decay process of free photoelectrons and shallow-trapped electrons in semiconductor crystals. The experimental results show that when the doping content is 10(-8)-10(-7) mol x mol(-1) Ag, the photoelectron decay process becomes slower, namely, the photoelectron decay time is longer, as the doping is near the grain surface before sensitization. After S+Au sensitization, the photoelectron decay becomes faster, showing that the sensitization centre acts as a deep electron trap. And when the doping is near the grain surface with 90% Ag, the photoelectron decay time becomes shorter, showing that the doping centre and the sensitization centre may interact. PMID- 17112022 TI - [Retrieval of monocyclic aromatic hydrocarbons with differential optical absorption spectroscopy]. AB - Differential optical absorption spectroscopy (DOAS) technique has been used to measure trace gases in the atmosphere by their strongly structured absorption of radiation in the UV and visible spectral range, e. g. SO2, NO2, O3 etc. However, unlike the absorption spectra of SO2 and NO2, the analysis of aromatic compounds is difficult and strongly suffers from the cross interference of other absorbers (Herzberg bands of oxygen, ozone and sulfur dioxide), especially with relatively low concentrations of aromatic compounds in the atmosphere. In the present paper, the DOAS evaluation of aromatic compounds was performed by nonlinear least square fit with two interpolated oxygen optical density spectra at different path lengths and reference spectra of ozone at different temperature and SO2 cross section to correct the interference from absorbers of O2, O3 and SO2. The measurement of toluene, benzene, (m, p, o) xylene and phenol with a DOAS system showed that DOAS method is suitable for monocyclic aromatic compounds monitoring in the atmosphere. PMID- 17112023 TI - [Spectroscopic measurements of atomic hydrogen concentrations in dielectric barrier discharge hydrogen plasmas]. AB - Atomic hydrogen plays important roles in chemical vapor deposition of functional materials, plasma etching, and surface cleaning. The present work introduces the fundamental principle to determine atomic hydrogen density via optical emission spectroscopy using Ar as an actinometer, and also reports the experimental results of atomic hydrogen density in the DBD discharge hydrogen plasmas. The variations of atomic hydrogen density and the hydrogen dissociation fraction as a function of pressure were calculated based on some of the available electron impact excitation cross section and quenching cross sections in the literatures. In this work, as the pressure increases from 0.32 to 5.1 kPa, the hydrogen dissociation fraction decreases from 5.2% to 0.089%, and the atomic hydrogen density decreases from 4.9 x 10(15) cm(-3) to 1.3 x 10(15) cm(-3). The variations of H atom Balmer lines and Ar (750.4 nm) emission intensity as functions of gas pressure, discharge voltage, and frequency were also investigated. PMID- 17112024 TI - [Study on concentration modulation spectra of the molecular ion beam by AC glow discharge]. AB - An experimental apparatus using an AC discharge was developed for producing molecular ion beam. The emission spectral intensity of N2+ is stronger than that of N2, and a very high ratio of N2+ (B)/N2 (C) up to 6:1, was achieved in N2 discharge. The effects and processes of N2 discharge, and the concentration modulation spectra were studied. The vibrational temperature and rotational temperature of N2+ molecular ions beam were calculated to be 3 310 and 282 K respectively from the emission spectra. PMID- 17112025 TI - [Study on interaction of anionic surfactant SDS and bovine serum albumin by fourier transform infrared spectroscopy]. AB - FTIR spectroscopy was applied to investigate the interaction of anionic surfactant Sodium Dodecyl Sulfate (SDS) and Bovine Serum Albumin (BSA). Amide band I of BSA was analyzed to obtain the change in secondary structure of BSA when different concentration of SDS was added and during different interaction period. In short interaction period and at low concentration of SDS, the alpha helixes increased and the random coil decreased. In long interaction period or at high concentration of SDS, SDS unfolded the protein by decreasing the alpha-helix structure and increasing the random coil. PMID- 17112026 TI - [Study on the identification of tea using near infrared reflectance spectroscopy]. AB - A rapid tea identification method by near infrared spectroscopy coupled with pattern recognition based on principal components analysis and Mahalanobis' distance technique was proposed. Four famous brand teas in China were studied, including Longjing tea, Biluochun tea, Maofeng tea and Tieguanyin tea in the experiment. In the spectral region between 6 500 and 5 300 cm(-1), through preprocessing method of MSC (multiplicative scatter comection), the prediction model was built. The result showed that the model was the best with 8 principal component factors. The rates of identification in calibration set samples and prediction set samples were 98.75% and 95%, respectively. A new idea about quick and precise identification of tea was offered. PMID- 17112027 TI - [Application of NIR quantitative analysis to nondestructive detection of loquat soluble solid content]. AB - The objective of the present research was to evaluate the potential of NIR diffuse reflectance spectroscopy as a way for nondestructive measurement of loquat soluble solid content of three varieties harvested from two orchards. According to the analysis, it was shown that the correlation coefficient of curves was relatively high in the two regions of 1400-1500 nm and 1900- 2000 nm. Statistical models were developed using partial least square regression (PLSR), stepwise multiple linear regression (SMLR) and principal component regression (PCR) methods in the full spectral region from 800 to 2500 nm and in the combined region of 1400-1500 nm and 1900-2000 nm. The results of PLSR model in the full spectral region were better than those of other models. The modeling results based on derivative spectra were not as good as those based on original spectra. Finally, a model was established based on the original spectra with 17 points smoothing for all the three varieties in the full spectral region by PLSR method. The correlation coefficients of calibration and validation were 0.96 and 0.95, respectively. The results indicate that it is feasible to use NIR spectroscopy technique for quantitative analysis of loquat soluble solid content. PMID- 17112028 TI - [Application of FTIR spectroscopy to the analysis of quality mensuration of Paeonia lactiflora Pall. from native habitat]. AB - In the present paper, different mensurations of Fourier transform infrared spectra for the analysis of Paeonia lactiflora Pall. were compared. FTIR mensurement was applied to the analysis of quality of xylem and outer layer of Paeonia lactiflora Pall. from native habitat. The results revealed that the chemical contents are the same in both the xylem and outer layer of Paeonia lactiflora Pall.. But outer layer has less paeoniflorin and its derivatives than in xylem of Paeonia lactiflora Pall. Different habitats of xylem of Paeonia lactiflora Pall.. were Hanzhou, Anhui and Sichuan. The results indicated that the quality with habitat in Zhejiang is the best among the three. PMID- 17112029 TI - [Calibration transfer used for the interpretation of RS-FTIR spectrum]. AB - A technique of analysis for RS-FTIR (Remote Sensing Fourier Transform Infrared) spectrum was built based on the principle of calibration transfer. A system of four-component gas mixtures, whose FTIR spectra were overlapped seriously, was determined as the object in the present research. Its FTIR data from EPA and field measurement were used to set up calibration and prediction models, respectively. With OSC (orthogonal signal correction), the model built with data of EPA predicted values of field measurement. The RMSEP values of acetone, methanol, benzene and chloroform were 0.0085, 0.0180, 0.0640 and 0.0028, respectively, while without OSC correction the corresponding values were 0.0856, 0.0479, 1.0653 and 0.0142, respectively. With optimization, the Kennard-Stone method was used for subset selection, and the iteration in the process of OSC was carried out 3 times, yielding the best performance. The result indicates that this technique can overcome the limitation of background and calibration model, which restricts the application of RS-FTIR in air monitoring. PMID- 17112030 TI - [Application of wavelength selection algorithm to measure the effective component of Chinese medicine based on near-infrared spectroscopy]. AB - Near infrared (NIR) spectroscopy has raised a lot of interest in the pharmaceutical industry because it is a rapid and cost-effective analytical type of spectroscopy with no need for extensive sample preparation, and with the easy realizable ability of on-line application. The NIR technology can increase the quality control standard of the Chinese medicine and accelerate the entry into the international market. In the present paper, two methods for wavelength selection are applied to the measurement of borneol, one of which is the multiple chain stepwise, which tends to select many variables in the same area containing valuable information, and the other is the mixture genetic algorithm, which incorporates simulated annealing so as to improve the local searching ability while maintaining the global searching ability. The results present that the number of wavelength is reduced to 16% compared with the original number of wavelength, and the prediction accuracy has increased 47.6%. Therefore, the method of wavelength selection is a good way to enhance the prediction accuracy and simplify the model in NIR region. PMID- 17112031 TI - [A new algorithm for generating Raman spectra using polychromatic light sources]. AB - In the present paper, the possibility of generation of Raman spectra using polychromatic light instead of laser is discussed based on the theory of Raman scattering. When a polychromatic light impinges on a sample, the resultant scattering includes Rayleigh scattering and Raman scattering brought about by the excitation light at each corresponding frequency. Mathematical analysis demonstrates that the scattering spectra after being corrected by the frequency of scattered light are the convolution of Raman-Rayleigh jointed spectrum and the power spectrum of the excitation light. Therefore, Raman spectrum can be obtained with an algorithm of Fourier transformation using polychromatic light as an excitation source. PMID- 17112032 TI - [Fluorescence spectrum of cesium vapor resonantly excited by the 852.3 nm laser line]. AB - The fluorescence spectrum of cesium vapor was studied in a cell irradiated with the light of 852.3 nm resonance line from a single mode laser diode. From the intensities and spectra of the Cs and Cs2 fluorescence we identified several collisional and radiative processes in the excited cesium atom-dimer system. The atomic lines of the highly excited states were the result of collisional energy transfer from Cs(6P 3/2) + Cs(6P 3/2) to Cs(6D, 8S) atoms. The Cs2 (B 1 IIu) band formed in Cs (6P) + Cs2 (X1sigmag+) collisions. The fine-structure mixing in 6 2P atoms was studied through excitation transfer, energy-pooling collision, and collisional excitation transfer between Cs2 and Cs. The authors estimated the value of rate coefficient for the collisional energy transfer from 6P3/2 to 6P1/2 to be (5.2+/-2.1) x 10(-11) cm3 x s(-1). For the excitation transfer process Cs2 (B 1 IIu)+ Cs(6S)--> Cs2 (X 1sigmag+) + Cs(6P1/2 ), a rate coefficient of (1.0+/ 0.4) x 10(-9) cm3 x s(-1) was yielded. PMID- 17112033 TI - [Study on the inclusion behavior of p-sulphonatocalix[4]arene with acridine by spectrofluorometric titrations]. AB - p-sulphonatocalix[4] arene (1) was prepared according to the literature, and spectrofluorometric titrations were performed to investigate the inclusion behavior of (1) and acridine in citrate buffer solution (pH 5.92, 0.1 mol x L( 1)) at different temperatures. It was found that in definite concentration range, the emission peak of acridine exhibited a slight red shift and th fluorescence intensity decreased when (1) was added. They form stable host-guest complex, and the stoichiometry of the inclusion complex is 1 : 1. The stability constants of the inclusion complex at 15.0 degrees C, 20.0, 25.0 and 30.0 degrees C were determined as 3.08 x 10(5), 4.45 x 10(4), 2.58 x 10(4) and 8.90 x 10(3), respectively. The thermodynamic parameters of inclusion process, deltaG, deltaH and deltaS, were determined. The experimental results indicated that the inclusion process was an exothermic and enthalpy-driven process. It was found that the stability constants descended when temperature rose. The most probable pattern of the inclusion complex between (1) and acridine was proposed as: acridine partially goes into the cavity of (1), and the protonated N atom and the negatively charged sulphonyl group bond firmly owing to strong electrostatic interaction. With the main contribution of electrostatic interaction and the assistance of Van de Waals and hydrophobic interaction, the host and the guest molecules form 1 : 1 supramolecular complex. PMID- 17112034 TI - [Study of the fluorescent response for optical ammonia sensing film]. AB - The characters of optical sensing films for ammonia were investigated based on a fluorescent indicator aminofluorescein (AF) immobilization. An organically modified silicates (ORMOSILSs) obtained by copolymerizing various proportions of methyltrimethoxysilane (Me-TriMOS) and tetramethoxysilane (TMOS) was applied. Compared with the fluorescent responses of fluorescein and fluorescein carboxylicate, the fluorescent intensity enhancement of AF for ammonia was found to be caused by the reaction of ammonia with the NH2 group on AF. The reaction may cause the reduction of the intermolecular self-quenching of AF, resulting in a fluorescence enhancement of sensing film in NH3 solution. Furthermore, the properties and the ammonia-sensing behavior of the film were investigated. The results presented here emphasized the significance of ormosil as a matrix for dissolved ammonia sensing. The detection limit for ammonia in water was 0.3 microg x mL(-1). PMID- 17112035 TI - [Study of fluorescence spectra of starch suspension]. AB - To achieve detection, monitoring and automation of starch graft copolymerzation, the investigation was carried out according to the luminescence of free radical produced in the reaction. The investigation on the fluorescence spectra of starch water suspension excited by ultraviolet light and its characteristics contributes to the study of the characteristics and mechanisms of free radical coming into being, emitting and disappearing. The experimental results indicate that starch water suspension can emit fluorescence, whose peak wavelength is about 339 nm. Furthermore, the physical mechanism of fluorescence emission of starch is analyzed. It is considered that this fluorescence comes from the transition from nonbonding electrons in the hetero-atom (O) of the functional group (C--O--C) called ether linkage to the antibonding orbital sigma*. PMID- 17112036 TI - [Fluorescence investigation on interaction between artemisinin (qinghaosu) and hemin and its analytical application]. AB - Hemin-catalytic decomposition of artemisinin (qinghaosu, QHS) was studied using pyronine B (PB) as an indicator. The interaction between hemin and QHS was an enzyme-substrate model, and the action sites were the endoperoxide moiety of QHS and the central metal ion of enzyme respectively. The kinetic catalytic constant depends upon enzyme and substrate concentrations, and the Michaelis-Menten parameters Km, Vmax and Kcat was 8.4 x 10(-5) mol x L(-1), 7.4 x 10(-6) mol x L( 1) s(-1) and 50.23 s(-1) respectively. The catalytic activity of hemin was inhibited in the presence of deactivated agents and at high temperature. Under optimal conditions, the change in fluorescence intensity (Fo-F) of pyronine B was proportional to the QHS concentration from 0.0 to 1.27 x 10(-6) mol x L(-1), and the detection limit (3sigma) was as low as 2.3 x 10(-8) mol x L(-1). The proposed method was applied to detect the concentration of QHS in the media of plasma and urine. PMID- 17112037 TI - [Synthesis, characterization and fluorescent properties of o, m, p (methacrylamido) benzoic acid]. AB - o, m, p- (Methacrylamido) benzoic acid (o, m, p-MAABA) was synthesized from o, m, p-aminobenzoic acid (o, m, p-ABA), elemental analysis and 1H NMR were used to confirm these compounds. The fluorescent properties of the compounds were investigated. The result showed that o, p-MAABA has lambda(ex)max at 358 nm while m-MAABA has lambda(ex)max at 324 nm. Excited at lambda(ex)max, o-MAABA had the strongest fluorescence, while p-MAABA had the weakest, and all of them were round at 420 nm. PMID- 17112038 TI - [Characterization of napropamide enantiomers by CD and determination of the enantiomeric ratios in water]. AB - The enantiomers of napropamide were separated by normal phase HPLC (HPLC: high performance liquid Chromatography) with Chiralpak OJ-H column and charactered by circular dichroism. On this basis, a method for the chiral separation and micro determination of napropamide in water was established. The linearity of calibration curve for racemic mixture was 10-100 ng x mL(-1) and the correlation coefficient was 0. 99. When 10 microL was injected, the detection limit of racemic mixture was 8 ng mL(-1), and the detection limits of both enantiomers were 4 ng x mL(-1). PMID- 17112039 TI - [The interaction of acridine orange with anionic surfactant and its application to the determination of protein]. AB - The change in the UV-absorption spectrum and fluorescence spectrum of acridine orange(AO) due to the addition of surfactant dodecyi benzene sulfonic acid sodium sait (SDBS) and bovine serum albumin(BSA) was studied. Meanwhile, the effects of the in situ formed AOAO dimer in SDBS as a fluorescence probe and BSA were discussed. A new method for the determination of BSA using fluorescence is presented. The results indicate that the method is sensitive and rapid. The linear range of determination is 0-4. 17 x 10(-7) mol x L(-1). The relative standard deviation is 1.9%, and the detection limit is 8.73 x 10(-10) mol x L( 1). PMID- 17112040 TI - [Selective determination for calf thymus DNA by a resonance light scattering technique with morin-Ce(IV) system]. AB - The interaction of DNA and the binary complex formed between Ce(IV) and morin was studied. With the maximum scattering peak located at 320 nm, the enhanced RLS was proportional to the concentration of ctDNA in the range 0-25 microg x mL(-1) and the detection limit (3sigma) of 0.3 microg x mL(-1) was obtained. Under the optimized conditions, four synthetic samples were determined with recoveries in the range 93.7%-108.4%. The proposed method is particularly attractive for its high selectivity for ctDNA in the presence of hsDNA by using certain sensitive spectrofluorometer since the intensity of RLS for hsDNA is far weaker than that for ctDNA. PMID- 17112041 TI - [Structure changes of lead silicate glasses induced by UV laser irradiation]. AB - The structure changes of lead silicate glasses induced by UV laser irradiation were studied by UV-visible spectra and electron spin resonance (ESR). The Urbach energies of lead silicate glasses exposed to the 266 nm UV laser increased. This implied that the disorder of the lead silicate glasses was increased. The electron spin resonance experiments showed that the oxygen deficiency centers in lead silicate glass did not change to paramagnetic defects after exposure to 266 nm laser beam, and the absorption peak near 235 nm for lead silicate glass film did not change either. But this absorption peak was bleached after exposure to the 248 nm UV laser. PMID- 17112042 TI - [UV spectroscopic characterization of the inclusion interaction of beta cyclodextrin and sym-diphenyl-thiourea and sym-diphenyl-urea]. AB - The inclusion interaction of beta-cyclodextrin and sym-diphenyl-thiourea and sym diphenyl-urea was studied by UV spectra. The stoichiometry ratio for the formation of the inclusion complexes was determined by Hildebrand-Benesi equation linear analysis and molar ratio method. The standard molar Gibbs energies, enthalpies, and entropies were derived for the inclusion process by Ks at different temperatrues. The result showed that the host:guest ratio of inclusion complex between the two diphenyl compounds and beta-CD is 2 : 1, the stability constant (Ks) of 2 : 1 inclusion complexes was higher than that of 1 : 1 inclusion complexes due to cooperative binding in the close two hydrophobic cyclodextrin cavities, and the association of the guest molecule with beta-CD was favored by enthalpy changes, proving that the Van der Waals interaction and the dipole-dipole interaction were main binding forces of cyclodextrin inclusion complex. PMID- 17112043 TI - [Studies on aclacinomycin spectral characters and the interaction between aclacinomycin and DNA by spectroscopic method]. AB - The interaction of aclacinomycin(ACR) and DNA was investigated by fluorescence spectrum, and the characteristics of the fluorescence and absorption of aclacinomycin (ACR) were studied. The results indicate that there are two situations: in the case that the concentration ratio of ACR to DNA is small, the ACR is intercalated into the stacked base pairs of DNA; in the other case that the concentration ratio of ACR to DNA is great, the interaction between ACR and DNA is complex. The binding constant of the interaction between ACR and DNA, calculated by the fluorescence titration method, is 2.7 x 10(6) mol x L(-1), and the binding site number is about 0.67 base pairs. PMID- 17112044 TI - [Interaction of quercetin and bovine serum albumin]. AB - The interaction of quercetin and bovine serum albumin (BSA) was investigated using fluorescence spectroscopy (FS) and ultraviolet spectroscopy (UV). The apparent binding constants (KA) between quercetin and BSA were 2.8 x 10(8) (26 degrees C) and 3.1 x 10(8) (36 degrees C), and the binding sites (n) were 1.7+/ 0.02. According to the Forster theory of non-radiation energy transfer, the binding distances (r) were also obtained. The experimental results showed that the quercetin could be inserted into the BSA, quenching the inner fluorescence by forming the quercetin-BSA complex. It was found that both static quenching and non-radiation energy transfer were the main reasons for the fluorescence quenching. The process of binding was a spontaneous molecular interactioln in which entropy increased while Gibbs free energy decreased, indicating that the interaction of quercetin and BSA was driven mainly by hydrophobic force. PMID- 17112045 TI - [Characteristics of absorption spectra of phytoplankton]. AB - Nine typical phytoplankton species were chosen and cultivated under two temperatures (20 and 15 degrees C) and two illumi- nations (7000 and 1100 lux), and their absorption spectra at different growth period were measured. Firstly, singular value decomposition was used on the matrix, which is composed of the autoscaled spectra data. The S1/msigma(i=1)Si was used to compare the similarities of spectra. Twenty-five representative spectra were obtained for the nine species of phytoplankton. Among them, there are one spectrum for each of Isochrysis galbana and Chaetoceros Debilis, two for each of Platymonas helgolanidica and Chaetoceros curvisetus, three for Gymnodinium sp., and four for each of Alexandrium tamarense, Prorocentrum dentatum, Skeletonema costatuma and Synechococcus sp. Then, feature extraction was processed to obtain the characteristic spectra, including seven incontinuous wavelengths that have great ability to differentiate species. They are 340. 5-420. 5 nm, 423. 5-431 nm, 440.5 525.5 nm, 760.5, 763.5, 769.5 and 856.5 nm. Consequently, they form characteristic spectra. The discriminant result is 80%. PMID- 17112046 TI - [Study on the structure and optical limiting properties of lead(II) tetrakis (4 cumylphenoxy)-phthalocyanine doped silica gel glass composites]. AB - The present paper reports the encapsulation of lead (II) tetrakis (4 cumylphenoxy)-phthalocyanine (PbPc(CP)4) into silica gel glass matrix to produce homogeneously doped composites by sol-gel technique. The existing state and structure of the doped PbPc(CP)4 were studied by UV-Vis absorption spectra and TEM images. Optical limiting properties were measured at 532 nm with 8 ns pulses. The results show that the doped PbPc(CP)4 molecules aggregate to form clusters in the gel glass composites, and PbPc(CP)4 has better optical limiting properties in the gel glass composites owing to the rigid structure of solid matrix. PMID- 17112047 TI - [Study on interaction between sulfonylurea herbicides and catalase by fluorescence spectroscopy]. AB - The binding of Sulfonylurea herbicides to catalase in aqueous solution was studied using fluorescence spectroscopy. It was shown that herbicides have a strong ability to quench the catalase fluorescence mainly through a static quenching procedure. The binding constant K and the number of binding site n were calculated according to the fluorescence quenching results. For chlorsufuron, K=8.69 x 10(5) L x mol(-1) and n = 1.16; for metsufuron methyl, K = 1.01 x 10(6) L x mol(-1) and n = 1.21; and for bensufuron methyl, K = 3.52 x 10(3) L x mol( 1), n = 0.77. It is clear that the binding of metsufuron methyl with catalase is stronger than that of chlorsufuron, while the binding of chlorsufuron stronger than that of bensufuron methyl. PMID- 17112048 TI - [Determination of antioxidation of the extract from Chinese medicine Morinda officinalis How by flow injection chemiluminescence and spectroscopy]. AB - Flow injection chemiluminescence analysis was used to determine the antioxidation activity of extract from Morinda officinalis How. The determination was based on the inhibition effect of Morinda officinalis How extraction in the chemiluminescence reaction of luminol - H2O2-CuSO4 system, and vitamin C was used as positive control. It was showed that Morinda officinalis How original liquid has obvious anti-radical activity. Spectroscopy was applied to determine the inhibition ratio of Morinda officinalis How extraction for the superoxide anion and hydroxy radicals in the special system. It was shown that the extraction from Morinda officinalis How could obviously scavenge superoxide anion and hydroxy radicals. It is suggested that the Chinese Medicine Morinda Officinalis How is a potential antioxidation activity medicine. PMID- 17112049 TI - [New approaches to selenium speciation]. AB - A review of the recent developments in selenium speciation analysis was presented, focusing on the techniques of separation, interface, elemental specific detection and identification of selenium species; the methods of sample storage and pretreatment storage for selenium speciation were also introduced. PMID- 17112050 TI - [Determination of trace germanium by HG-ICP and study on the medium influence]. AB - A method to determine the content of trace germanium by hydride generation inductively coupled plasma atomic emission spectrometry was established. The influences of different acidic medium, concentration of NaBH4 and other concomitant elements are examined systemically. The optimal analytical conditions were discussed and examined. The detection limit is 0.2 ng x mL(-1) and the precision is 0.9%. The value of germanium content detected in environmental reference material agrees with the certified value. The medium influence on determining trace germanium was studied in detail. PMID- 17112051 TI - [Effect of nebulizer gas pressure on the emission intensity of ICP]. AB - In an attempt to inhance the emission intensity of ICP, a conventional concentric pneumatic nebulizer was used with increased pressure, and the aqueous aerosol was partially let out to modulate the flow velocity of sample solution into the flame of the ICP. In the present work, the emission spectra of elements Ca, Si, Sr and Zn in water samples were measured, and the results showed that corresponding emission intensities are the highest at 0.05 MPa under the conventional condition, and after increasing the nebulizer gas pressure to 0.07 MPa and letting out part of aqueous aerosol, the peak values grew by 38%, 79%, 45% and 70%, respectively. In addition, the stability of the plasma radiation was not affected by the higher nebulizer gas pressure. PMID- 17112052 TI - [Determination of elements related to reducing blood sugar (ERBS) in tea and tea polysaccharide by ICP-MS]. AB - Contents of elements related to reducing blood sugar (ERBS) in several kinds of green tea of Wuyuan county, their tea infusion extracted by warm water and boiling water and tea polysaccharide (TP) were determined by ICP-MS, after the samples were digested with acids by microwave method. The relation between the amount of ERBS and tea quality and the relation between the content of ERBS in TP and the total content in tea were discussed. The results showed that the amounts of ERBS in tea and TP are different in various samples. The amount of ERBS in TP extracted from coarse tea was higher than that from fresh tea. The percentage of certain ERBS in TP accounting for the ERBS in tea ranged from 0.03% to 9.57%. As to the total ERBS in TP extracted from Mt. Zhang super grade tea and grade 5 tea, the percentages were 1.11% and 2.10% respectively. As to Mt. Gu old tea, it was 0.85%. The results could provide data for selecting tea material used to extracting tea polysaccharide and for studying the mechanism of reducing blood sugar. PMID- 17112053 TI - [Analysis of the degrading products of carbofuran by Sphingomonas sp. with GC/MS and GC/FTIR]. AB - In different degrading phases, the degrading products of carbofuran by CDS 1(Sphingomonas sp. ) were extracted by acetone. According to the analysis of gas chromatography/mass spectrometry (GC/MS), carbofuran-7-phenol was identified as the initial degrading product, and an unknown metabolite with molecular weight of 182 was determined as 2-hydroxyl-3-tertiarybutylalcohol-phenol, which was the next degrading product of carbofuran-7-phenol. According to the analysis of GC/MS and gas chromatography/Fourier transform infrared spectrum (GC/FTIR), the main component of the compounds that appeared in the final phase of degradation which could produce volatile pungent odor, was identified as carvone. PMID- 17112054 TI - [Determination of 27 microelements in 11 shellfishes]. AB - Shellfish samples were prepared by microwave, and 27 microelements including Ca, Mg, K, Na, P, S, Zn, Mn, Fe, Cu, Cr, Co, Se, Sn, Ni, Al, Ti, V, Mo, Ba, Ge, Si, Cd, I, Pb, As and Hg were determined by ICP-AES. It was showed that ICP-AES had an obvious advantage in the determination of microelements, and shellfishes contained upwards of 20 kinds of microelements. Calcium in fresh water was higher than that in seawater, and As and Pb were the major pollutants. PMID- 17112055 TI - [Atomic emission spectrometry determination of Au, Pt and Pd after separation and enrichment by hyperbranched polymer]. AB - The present paper shows that the trace amount of gold, platinum and palladium in hydrochloric acid solution can be concentrated by hyperbranched polymer. The new reagent has a rapid adsorption rate and big concentrating capacity. The determination of trace Au, Pt and Pd in sample using carbon powder and strontium carbonate as buffer was carried out by atomic emission spectrometry(AES). Zirconium was selected as internal standard line. The sample was directly loaded into ordinary electrode. The method is simple, rapid and accurate. The condition of determination, and factors of influence were studied. The analysis line of Au, Pt and Pd is 312.3, 306.5 and 311.4 nm respectively. The internal standard line of Zr is 310.7 nm. The linear range of the determination of Au, Pt and Pd is 0-0. 20%, 0-0. 40% and 0-0. 20% respectively. The detection limit of Au, Pt and Pd is 0.010%, 0.0030% and 0.0030% respectively. The method has been applied to the determination of Au, Pt and Pd with satisfactory results. PMID- 17112056 TI - [Determination of 15 trace elements in antineoplastic traditional Chinese medicine by atomic absorption spectrometry]. AB - Five kinds of antineoplastic traditional Chinese medicines including fufangtianxian capsule, xiaoaiping tablet, zhiling capsule, shenlian capsule and shenlinglan capsule were digested with HNO3-HCl mixed acid. The fifteen trace elements including calcium, magnesium, potassium, sodium, iron, zinc, manganese, cobalt, nickel, chromium, silver, copper, strontium, cadmium and lead in the above-mentioned drugs were determined by atomic adsorption spectrometry. The effects of the kinds of mixed acid, the ratio of HNO3 to HCl in the mixed acid, the volume of digesting solution and the digesting time were also investigated in detail. The results obtained show that the concentrations of Ca, Mg, K, Na, Fe, Zn, Mn, Co and Ni in the five kinds of antineoplastic Chinese traditional medicines are higher than those of other elements. PMID- 17112057 TI - [Determination of trace lead and iron in nickel chloride and manganese sulfate by flame atomic absorption spectrometry after coprecipitation with yttrium phosphate]. AB - Using yttrium phosphate as the coprecipitation collector for the separation and preconcentration of trace lead and iron in nickel chloride and manganese sulfate, flame atomic absorption spectrometric (FAAS) determination was described in the present paper. Coprecipitation parameters including the pH of the solution, and the amounts of YCl3 and H3 PO4 were discussed. It was found that lead and iron in nickel chloride could be coprecipitated quantitatively in the range of pH 3.0 4.0, and so could be lead in manganese sulfate. The detection limits (3sigma) of lead and iron in 20 mL solution were 1.63 x 10(-2) mg x L(-1) and 4.58 x 10(-2) mg x L(-1) respectively. In NiCl2 solution the standard addition recoveries for lead and iron were 100.91% and 99.73% respectively, and in MnSO4 solution the standard addition recoveries were 99.45% and 98.98% respectively. The method has eliminated the interference of matrix, and the result is satisfied. PMID- 17112058 TI - [Determination of trace elements in Spirulina platensis (Notdst.) Geitl. by flame atomic absorption spectrometry combined with microsampling pulse nebulization technique]. AB - The contents of trace elements Ni, Zn, Mn, Cu, Mg, Fe, Ca and Pb in Spirulina platensis (Notdst.) Geitl. were determined by flame atomic absorption spectrometry combined with microsampling pulse nebulization technique. The results of the determination show that Spirulina platensis (Notdst.) Geitl. are rich in the inorganic elements such as Mg, Zn, Fe, Ca and Cu. Its recovery ratio obtained by standard addition method ranged between 96.58% and 106.12%, and its RSD was lower than 4.26%. The result will provide scientific data for the study on the trace elements in Spirulina platensis (Notdst.) Geitl. and on their relativity of efficacy of medicine. PMID- 17112059 TI - [A method for obtaining redshifts of quasars based on wavelet multi-scaling feature matching]. AB - The LAMOST project, the world's largest sky survey project being implemented in China, is expected to obtain 10(5) quasar spectra. The main objective of the present article is to explore methods that can be used to estimate the redshifts of quasar spectra from LAMOST. Firstly, the features of the broad emission lines are extracted from the quasar spectra to overcome the disadvantage of low signal to-noise ratio. Then the redshifts of quasar spectra can be estimated by using the multi-scaling feature matching. The experiment with the 15, 715 quasars from the SDSS DR2 shows that the correct rate of redshift estimated by the method is 95.13% within an error range of 0. 02. This method was designed to obtain the redshifts of quasar spectra with relative flux and a low signal-to-noise ratio, which is applicable to the LAMOST data and helps to study quasars and the large scale structure of the universe etc. PMID- 17112060 TI - [Transient dynamics of excited states and nonlinear optical properties of InP nanoparticles]. AB - The decay curves of the emission at different wavelength were measured. One nanosecond of transition lifetime of interband was obtained. The transient dynamics of InP nanoparticles was investigated by one color femtosecond pump probe method at the wavelength of 800 nm. The experimental result shows that the saturation of exciton resonance results in photobleaching at 800 nm The decay process of the bleaching includes two components, i.e. the fast one from free carries scattering and the slow one from trapped carriers scattering. The nonlinear optical properties of InP nanoparticles were investigated by using femtosecond optical Kerr effect. The magnitude of chi3 for InP nanoparticles embedded in SiO2 sol-gel glass was calculated, and its origin was analyzed. PMID- 17112061 TI - [The research on preparing CeO2 nanocrystalline by homogeneous precipitation method]. AB - CeO2 nanocrystallines were prepared by homogeneous complexed-precipitation method, using cerous nitrate and ammonium tartrate as raw materials. The effects of cerous tartrate complex compound and the way of producing precipitation on the particle size of samples were investigated. The samples were characterized by XRD, TEM and SEM. The SEM micrograph shows that the foam exhibits a perforated porousness stereostructure in shape, and the HRTEM picture of the particles reveals the clear crystal lattice. All the results indicate that the samples were CeO2 nanocrystalline. PMID- 17112062 TI - [Nitrogen stress measurement of canola based on multi-spectral charged coupled device imaging sensor]. AB - Site-specific variable nitrogen application is one of the major precision crop production management operations. Obtaining sufficient crop nitrogen stress information is essential for achieving effective site-specific nitrogen applications. The present paper describes the development of a multi-spectral nitrogen deficiency sensor, which uses three channels (green, red, near-infrared) of crop images to determine the nitrogen level of canola. This sensor assesses the nitrogen stress by means of estimated SPAD value of the canola based on canola canopy reflectance sensed using three channels (green, red, near-infrared) of the multi-spectral camera. The core of this investigation is the calibration methods between the multi-spectral references and the nitrogen levels in crops measured using a SPAD 502 chlorophyll meter. Based on the results obtained from this study, it can be concluded that a multi-spectral CCD camera can provide sufficient information to perform reasonable SPAD values estimation during field operations. PMID- 17112063 TI - [Simultaneous determination of total flavone and total saponin in Gynostemma pentaphyllum by signal multiplier spectrophotometry]. AB - Important chemical constituents analysis for the total flavone and total saponin in Gynostemma pentaphyllum is described. The colour reactions of flavones and saponines with vanillin-perchloric acid in acetic acid produced the good absorptions at 451 and 547 nm, but the absorption peaks too overlapped to be determined simultaneously. A new method for the total flavone and the total saponin in Gynostemma pentaphyllum to be determined by signal multiplier spectrophotometry simultaneously without any preliminary separation was proposed. For quantitative analysis, the rutinum as a standard of the total flavone and the ginsenoside Rb1 as standard of the total saponin were applied. The experiment results showed that the regression equations of concentration and deltaA were obtained: deltaAflavone = 0.0133+4.417 0Cflavone, relation coefficient rflavone = 0.9994, and the total flavone concentrations were in 0-0. 16 microg x mI(-1) with deltaA obeying linear relation; deltaAsaporin = 2.775 5Csaponin -0.8881 x 10(-2), relation coefficient rsaponin = 0.9991, and the total saponin concentrations were in 0-0.30 microg x mL(-1) with deltaA obeying linear relation respectively. The recovery ratio was 104.0%-113.0% and 86.8%-94.6% respectively. The RSDflavone was less than 0.58% (n = 9) and RSDsaponin was less than 0. 35% (n=9) respectively. The proposed method is simple, rapid accurate and feasible. PMID- 17112064 TI - [Comparison of light attenuation characteristics and optical penetration depths between native and coagulated human liver tissues]. AB - A double-integrating-spheres and IAD method were used to study the differences in the optical penetration depths (OPDs) and light attenuation (LA) native and coagulated human liver tumors and liver tissues at the wavelengths of 680, 720, 780, 810, 850 and 890 nm of Ti: Sapphire laser. The results of measurement showed that the OPDs for native and coagulated human liver tumors and liver tissues at six different wavelengths obviously increase with increasing laser wavelength, the OPDs of coagulated human liver tumors and liver tissues at six different wavelengths were significantly smaller than that of native human liver tumors and liver tissues at the same wavelength respectively (P<0.05), and the OPDs of native and coagulated human liver tumors at six different wavelengths were significantly bigger than that of native and coagulated human liver tissues at the same wavelength respectively (P<0.05). The LA for native and coagulated human liver tumors and liver tissues at six different wavelengths obviously decreases with increasing laser wavelength, and the LA for coagulated human liver tumors and liver tissues at six different wavelengths is significantly bigger than that for native human liver tumors and liver tissues at the same wavelength respectively (P<0.05). The LA for native and coagulated human liver tumors at six different wavelengths is significantly bigger than that for native and coagulated human liver tissue at the same wavelength respectively (P<0.05). PMID- 17112065 TI - [The real-time FOCSDT detection method and conditions for dissolution test of metronidazole and vitamin B6 tablets]. AB - A method to monitor the dissolution rate of drug on-line fiber optic chemical sensor dissolution test(FOCSDT) was studied. Bifurcated optical fiber was used to connect the light source and detector, and the common end was dipped in the dissolution vessel. The dissolution process could be monitored through computer. The results show that the high, middle and low concentration group's recovery of metronidazole is 100.8%, 99.8% and 100.6%; and RSD is 2.5, 0.8 and 1.1 respectively, The high, middle and low Concentration group's recovery of vitamin B6 is 98.8%, 100.8% and 98.8%; and RSD is 4.1, 4.1 and 2.5 respectively. This process of analysis can test the whole process of drug and get dissolution graph. The study shows that this process analysis can reflect the real dissolution of drug and obtain the total information. PMID- 17112066 TI - [Determination of writing age of blue ballpoint pen inks by high performance liquid chromatography]. AB - As it is a frequently encountered problem in the laboratory of forensic science nowadays to distinguish whether the questioned documents, such as deeds, contracts, and receipts, written in ballpoint pen inks are true or not, and identify the writing age of them, it is very essential to establish a simple, sensitive and accurate method to examine the similarities and differences of the ballpoint pen inks and identify the writing age. The present paper introduces a technique that allows identifying the kind and the writing age of the blue ballpoint pen inks. The technique is based on using a high performance liquid chromatographic method for distinguishing the similarities and differences in dyes of blue ballpoint pen inks and determining changes in dyes of blue ballpoint pen inks developed with age, and these changes can be evaluated by the ratio of peak areas. PMID- 17112067 TI - [Microbiochemical analyzer based on continuous spectrum and its test for clinic use]. AB - A microbiochemical analyzer based on continuous spectrum and its test for clinic use are introduced. The principle of splitting behind was adopted to design the micro biochemical analyzer, which has the characteristics of little disorder light, real time collection of continuous spectrum signal (340-770 nm), small volume, light weight, small wastage of sample and reagent and rapidness and direct view of check. It has wide application in the emergency treatment, middling and small hospitals, rescue of battleground, and medicinal research. Representative items for clinic test were selected, such as uric acid, total cholesterin and albumin in our experiments. Comparative test and analysis of the daily check-up samples were made respectively by our microbiochemical analyzer and by the standard one of the hospital (Olympus AU2700, RT-1904C, Beckman LX 20). The test results show that our micro biochemical analyzer can meet the requirement of clinic use. PMID- 17112072 TI - Draconian industrial laws hit Australian workers hard. PMID- 17112073 TI - Providing choice and expertise in sexual and reproductive health. Interview by Anne Manchester. PMID- 17112074 TI - The role of the nurse in oral health. PMID- 17112075 TI - Respecting breastfeeding choices. PMID- 17112076 TI - Forum tackles non-communicable diseases. PMID- 17112077 TI - Understanding the competence review process. PMID- 17112078 TI - Election planning for the next Nursing Council in 2009. PMID- 17112079 TI - Negotiating from a position of interests rather than claims. PMID- 17112080 TI - Overcoming Maori women's resistance to cervical smears. PMID- 17112081 TI - [Hospital internist: profession with a future or endangered species?]. PMID- 17112082 TI - [Alternative to hospitalization for patients diagnosed with acute congestive heart failure]. AB - Heart failure is the first cause of hospital admission in patients above 65 years of age and represents a major health problem given demographic projections. Rapid admission of selected patients in specialized heart failure observation units in the emergency departments providing therapeutic monitoring and multiple educative interventions may obviate the need for hospitalization. This strategy may reduce the overall incidence of hospitalization and subsequent readmissions as well as save costs. Outcome studies are needed before a widespread implementation of such strategies. PMID- 17112083 TI - [Impact of urinary incontinence in community-dwelling women]. AB - Urinary incontinence (UI) is a frequent problem in an aging population and prevalence of urinary incontinence for elderly women may be up to 50% and imposes a huge burden on the health care system comparable to osteoporosis or osteoarthritis. The impact of UI shows clear associations between UI and depression and decline of quality of life comparable to congestive heart failure, major respiratory condition or gastrointestinal tract conditions. Some limited data suggest that UI is also a risk factor for institutionalization and hospitalization. Despite effective treatments, UI is significantly underdiagnosed by clinicians, and underreported by patients. This results in unmet needs for incontinence treatment, in particular for older individuals. PMID- 17112084 TI - [Primary prevention of gastroduodenal ulcer: what is the evidence?]. AB - Primary prevention of gastroduodenal ulcer and its major complication, gastrointestinal bleeding, has been studied mainly in the intensive care environment. Proton pump inhibitors (PPIs), H2-receptor antagonists and sucralfate have proved effective. By extension, PPIs are inappropriately prescribed in a variety of clinical situations. In contrast, they are probably underused in patients chronically treated by nonsteroidal anti-inflammatory drugs. This article reviews the situations in which an increased ulcer risk justifies primary prevention. Validated prophylactic options are also addressed. PMID- 17112085 TI - [Deployment of a computerized physician order entry: description of the process and challenges]. AB - The introduction of a system of computerized physician order entry (CPOE) decreases medication error rates by formalizing the drafting of the orders and providing alarms which announce potential side effects, drug interactions and overly high doses. The quality of the CPOE does not insure its success. Its deployment must be preceded by an assessment and update of the available computer resources, by training of the users and setting up a help-desk attainable 24 hours a day. The project must be lead by senior medical and nursing staff. A joint effort is the key to ensure the success of CPOE and facilitate its acceptability. CPOE provides opportunities for improving: patient safety, training of professionals, quality and efficiency of care. CPOE systems are an ideal basis to set up clinical protocols and clinical pathways. PMID- 17112086 TI - [Anticoagulants and cancer: beyond thromboembolic disease prevention]. AB - The association between cancer and thromboembolic disease is well documented. The procoagulant capacity of tumor cells seems to be related to malignant transformation. In this context, a preventive effect on tumoral progression by the adjunction of an anticoagulant such as vitamin K antagonists, unfractionated heparin or low molecular weight heparin (LMWH) was studied. Some studies concluded to a modest increase in survival in patients treated by LMWH in adjunction to conventional chemotherapy and/or radiotherapy. This benefit seems to be related to an inhibitory effect on tumoral progression. LMWH seems promising in an eventual routine clinical use. However, their type, dose and duration and in which cancers has to be further defined. PMID- 17112087 TI - [Perinatology for the pediatrician]. AB - From the standpoint of the pediatrician, the new knowledges in perinatology allowed progresses in certain fields (identification of high risk pregnancies, decrease in perinatal mortality, decrease of major handicaps in high risk newborns). However, the new knowledge's did not improve the rate of preterm deliveries. Some aspects of antenatal and intrapartum fetal assessment as well as the postnatal evaluation of the newborn will be discussed. The figures and tables summarize data directly linked to the practitioner's every day's concerns. PMID- 17112088 TI - [Working conditions regulated by in-training Swiss physicians: one year of experience]. PMID- 17112089 TI - [Robotic orthoses in French-speaking Switzerland to rehabilitate paraplegic patients]. PMID- 17112090 TI - [Marketing makes pressure! ACE inhibitors or AT2? We must choose...]. PMID- 17112091 TI - [When The Lancet invites itself to Geneva, Appia Avenue]. PMID- 17112092 TI - [Pregnancy and nicotine substitution treatments: what to do?]. PMID- 17112093 TI - [Human and painful embryologic chimeras]. PMID- 17112094 TI - [The long hospital pathway]. PMID- 17112095 TI - On membership, unity, and playing nice in the sandbox. PMID- 17112096 TI - Florida's emergency health volunteer registry (FEHVR). PMID- 17112098 TI - The need for recurring funding of SUS nursing schools: a FLU student's perspective. PMID- 17112099 TI - Nursing students as lobbyists: our Tallahassee experience. PMID- 17112100 TI - Mirror, mirror on the wall, what's in my personal care products: a short article about 300,000,000 "guinea pigs". PMID- 17112101 TI - Nursing and all of its possibilities and what the future is bringing. PMID- 17112102 TI - Am I allowed to do that? PMID- 17112104 TI - Adequate access to health care on the line. PMID- 17112105 TI - Influenza. PMID- 17112106 TI - Fundamental technical error. PMID- 17112107 TI - AMS Physicians Health Committee & the Arkansas Medical Foundation. Helping physicians (and others) reclaim their lives. PMID- 17112108 TI - Certification commission setting standards for HIT. PMID- 17112109 TI - Physiatry. Understanding and utilizing this unique specialty. PMID- 17112110 TI - Myxedema coma in a patient with Down's syndrome. AB - INTRODUCTION: hyroid dysfunction is common in Down's syndrome, most common being hypothyroidism. Longstanding, untreated hypothyroidism can lead to myxedema coma. METHODS: Here we report a patient with Down's syndrome who presented with myxedema coma. DISCUSSION: The three essential elements for the diagnosis of myxedema coma include altered mental status, defective thermoregulation and a precipitating event or illness; all of these were present in our patient. Also, very high TSH, low T3 and T4, and the rapid response to the treatment with levothyroxine confirmed the diagnosis. CONCLUSION: Patients with Down's syndrome should have regular screening for thyroid dysfunction. PMID- 17112111 TI - Effect of gender on pain perception and pharmacologic management. PMID- 17112112 TI - Where do they come from? How do they find me? PMID- 17112113 TI - All the difference in the world. PMID- 17112114 TI - The work of the world. PMID- 17112115 TI - Coast to coast, or dental: mental health for dummies. PMID- 17112116 TI - Part one: The restoration of non-vital teeth: structural, biological, and micromechanical issues in maintaining tooth longevity. AB - This manuscript will review research from 1967 to the present to find the best evidence for the reconstruction of non-vital teeth. The paper will review the contention that non-vital teeth are "more brittle" by analyzing the relevant physical properties of vital versus non-vital teeth; describe the structures of the tooth used to manage stress and strain; describe the formation of dentinal cracks and propagation of fracture planes through dentin; and analyze forces placed on human teeth and their effects on the tooth in Part One. In Part Two, the paper will describe the strength of various dowel and core designs relative to strength, retention, and durability and describe the results of testing various dentin bonding materials in strength and retention of dowels in non-vital teeth. The review will conclude with recommendations as to the materials and techniques in specific clinical situations best capable of single tooth reconstruction to ensure tooth longevity together with the scientific basis for their adoption. PMID- 17112117 TI - Part two: Trends in Minnesota dental practice. AB - As the practice of dentistry continues to evolve, the wise practitioner will make steady, thoughtful improvements on all three fronts: clinical, technological, and managerial. These surveys help give the practicing dentist the perspective he or she will need to make the plan that will suit an individual office. PMID- 17112118 TI - What's a dentist to do? Keeping your balance. PMID- 17112119 TI - Real value, or just cheap? PMID- 17112120 TI - We must all join together now: tort reform will not save us if we have no revenue. PMID- 17112121 TI - Healthy Maryland Initiative. PMID- 17112122 TI - Politics and promises. PMID- 17112123 TI - Some reflections on health care reform. PMID- 17112124 TI - Fair Share Health Care. PMID- 17112125 TI - A lawyer's perspective: Retail Industry Leaders Association v. James D. Fielder, Jr., Maryland Secretary of Labor. PMID- 17112126 TI - Insuring Maryland's uninsured. PMID- 17112127 TI - Health care reform in Massachusetts--a work in progress. PMID- 17112128 TI - Hurricanes provide impetus for health system reform in Louisiana. AB - The clear lesson, for Louisiana and any other state that is contemplating the potential disruption of health care following large-scale dislocations caused by either natural or man-made major disasters, is that proper mechanisms must be put in place before the event. This is necessary to provide greater portability of health care coverage and alternatively, temporary finance that coverage. Our Louisiana State Medical Society's plan Health Access Louisiana will help Louisiana recover from the devastating effects of Hurricanes Katrina and Rita and serve as a model for the reform of our healthcare coverage system for our country. We firmly believe the devastation in Louisiana presents a unique opportunity to rebuild a healthcare system from scratch. The new system will not be a modification of the old system, which did not work, but a system which effectively and economically offers equal access to high quality healthcare for all. PMID- 17112129 TI - Maryland gubernatorial candidates present health care platforms. PMID- 17112130 TI - Position paper on health-related issues. PMID- 17112131 TI - Get your ACTivities together: New CMS regulations in long-term care. PMID- 17112132 TI - Conflicting views about codependency. PMID- 17112133 TI - Evidence-based guideline: Promoting spirituality in the older adult. PMID- 17112134 TI - Providing quality care in assisted living facilities: Recommendations for enhanced staffing and staff training. PMID- 17112135 TI - Assessing the stability of values and health care preferences of older adults: A long-term comparison. AB - The purpose of this study was to examine the stability of health care decisions and the influencing values of competent older adults in late life (N = 21) during a 7-year period. Participants were given a list of value indicators from which they were asked to identify the five that most influenced their health care decision-making process. They also were instructed to make decisions related to five health care treatment scenarios and five life-sustaining treatment scenarios across two health states and three different levels of prognosis. Face-to-face interviews were conducted initially followed by telephone interviews 7 years later. No significant change in health care decisions was found for life sustaining treatments during the 7-year period in study participants. The only significant change between the initial and telephone interviews for health care treatments was an increase in the number of participants not consenting to hip replacement surgery regardless of health state or prognosis. Similarly, change occurred between the initial and telephone interviews with more participants refusing cataract surgery when the prognosis was a 50% chance of recovery. Stability also was noted in the values selected by participants. Four of the top five values selected at the initial interview also were selected 7 years later. PMID- 17112136 TI - Promoting successful cognitive aging in adults with HIV: strategies for intervention. AB - Aging individuals with HIV may be at risk for developing more age-related cognitive decline, mild cognitive impairment, or even dementia. The incidence of HIV-related dementia was reduced sharply with the introduction of HAART. Positive mediators, such as good nutrition or cognitive remediation therapy, can potentially mitigate some of the negative cognitive consequences of aging with HIV. PMID- 17112137 TI - Examining the role of gender in health-related quality of life: Perceptions of older adults with chronic obstructive pulmonary disease. AB - The purpose of this cross-sectional study was to explore the impact of gender on health-related quality of life perceptions among 67 older adults with chronic obstructive pulmonary disease who were living with a spouse. Physical and psychosocial health-related quality of life perceptions as well as demographic and illness-related variables were compared by gender. A correlation and regression analysis revealed women experienced significantly greater psychosocial impairment, while preliminary support was found for symptom-related impairment among men. Initial recommendations for nursing interventions include incorporating anticipatory guidance from older adults with late stages of chronic obstructive pulmonary disease as well as exploring the impact of symptoms. PMID- 17112138 TI - The shortened food expectations--Long-term care questionnaire: Assessing nursing home residents' satisfaction with food and food service. AB - Lack of nursing home resident satisfaction with meals often results in reduced food intake, leading to poor nutritional status, weight loss, functional decline, and depression. The purpose of this article is to describe the development and initial testing of the 28-item revised Food Expectations-Long-Term Care (FoodEx LTC) questionnaire with a convenience sample of nursing home residents (N = 61). Because of possible respondent burden, the original 44-item, five-domain FoodEx LTC was revised, resulting in the deletion of 16 redundant items and those with inter-item correlations less than .25. Coefficient alpha scores ranged from .65 to .82, and test-retest correlations ranged from .79 to .88, dependent on domain. This revised instrument has good initial validity and reliability, resulting in a shorter instrument that accurately assesses nursing home resident satisfaction with food and food service. PMID- 17112139 TI - Kentucky diabetes trend data reveals strong progress yet room for improvement as flu season approaches. PMID- 17112140 TI - Meeting the health care needs of medically underserved, uninsured, & underinsured Appalachians. AB - There is the need for critical changes in the United States health care system to meet the rising costs of health care and the massive number of people without health insurance. The Berea Health Ministry Rural Health Clinic (BHMRHC) is a nurse-managed, rural primary health care clinic that was designed to respond to the health care needs of the underserved, uninsured, and underinsured populations in a designated nine county area of Appalachian Kentucky. The process used in the development and implementation of BHMRHC is described as a model to assist in the future development of similar clinics. PMID- 17112141 TI - Magnet designation awareness. Our role as nursing faculty. PMID- 17112142 TI - Managed care versus managed "carelessly" the nurse case manager's role in the managed care arena. PMID- 17112143 TI - Data bits. What your heart wants you to know: risk symptoms in older adults. PMID- 17112144 TI - KNA long-term care program. PMID- 17112145 TI - Pay rise proposal is 'slap in the face'. PMID- 17112146 TI - "In 21st century Britain 1.8 million pensioners live in poverty". PMID- 17112148 TI - What's the future for community nurses? PMID- 17112147 TI - "Children's palliative care services are at the mercy of an insecure future". PMID- 17112149 TI - Helping children overcome deafness. PMID- 17112150 TI - Guidelines on the management of peripheral arterial disease. AB - Last week guidelines on the diagnosis and management of peripheral arterial disease were published by The Scottish Intercollegiate Network. This article discusses the recommendations and highlights implications for practice. PMID- 17112151 TI - Chest examination, part 1--chest palpation. PMID- 17112152 TI - Respiratory assessment as part of track and trigger. AB - The respiratory system's primary function is to provide oxygen to all the cells of the body for metabolism and to eliminate the by-product of this metabolism, carbon dioxide (Tortora and Grabowski, 2002). In this article (part three of four) we aim to explore respiratory rate and related care. PMID- 17112153 TI - International consensus on managing lymphoedema. AB - Philip Morgan and Christine Moffatt outline the reasons why an international consensus document on the management of lymphoedema is needed and explain how the document was developed. PMID- 17112154 TI - Choosing an appropriate antibacterial dressing. AB - There have been significant developments in the dressings available to treat infected wounds; this increased choice can, however, be confusing. Jacqui Fletcher outlines the key considerations for using an antimicrobial dressing and describes the key dressing groups that can be used to treat wound infection. PMID- 17112155 TI - Caring for a patient with malodorous leg ulcers. AB - Patients with leg ulceration who experience symptoms of pain and malodour can become isolated and suffer psychological problems. Peter Cooke describes a patient who experienced these problems and identifies the importance of patient involvement in planning care. PMID- 17112156 TI - Assessing mixed venous and arterial leg ulcers. AB - Caroline Dowsett outlines the key points in the assessment and management of mixed venous and arterial leg ulcers so that readers can consider their own practice against expert opinion. PMID- 17112157 TI - "In the future we won't have any nurses". PMID- 17112158 TI - [Image of the month: Kaposi disease in an African AIDS patient]. PMID- 17112159 TI - [Biermer disease disclosed by mucous ulcerations]. AB - Pernicious anemia rarely induces mucous membranes lesions, wich, exceptionally reveal the disease. We report a case of a 64-year-old man who presented a clinical history of recurrent painful ulcerations of the buccal, genital and ocular muosa. He also had peripheral sensory nerve impairement of the legs. The diagnosis of pernicious anemia was made after a 19 year course. Cobalamin supplementation induced an improvement of the mucous membrane lesions and of the anemia, but recurence occurred each time the treatment was stopped. Our observation is particular by the multiple mucous membrane involvement revealing pernicious anemia and by the ocular lesions which had never been previously reported. PMID- 17112160 TI - [Pedunculated and pigmented basal cell carcinoma: an unusual presentation]. AB - Basal cell carcinomas (BCC) are the most common malignant neoplasms in humans. Clinical misdiagnoses are not uncommon. These cancers are classified according to their histological and clinical characteristics which exhibit distinct malignant behaviours. It is important to be aware of the different clinical presentations both for the accurate diagnosis and tumour management. We report the case of a 78 year-old man, who presented an unusual pedunculated and pigmented BCC. This combined aspect is rarely encountered. PMID- 17112161 TI - [Increasing cost of insulin therapy in Belgium. From a critical analysis of the situation to a search for practical solutions]. AB - Cost related to insulin therapy is markedly increasing in Belgium, as in other Eucopean countries. In the present paper, we will briefly analyze the main reasons for such aa increase, integrate such observation withIn the global context of diabetes management and suggest some solutions to provide best care to insulin-treated diabetic patients at a reasonable cost. The rise of the cost of insulin therapy has a multifactorial origin. It mainly results from an increase in the number of diabetic patients, a more intensive management, In both type 1 and type 2 diabetes, and a greater use of more expansive insulin analogues. It is important to analyze the increase of the cost of insulin therapy within the global burden of diabetes melitus. Only a better responsibility of all health care partners, patients, physicians, pharmaceutical companies, public health authorities, could provide solutions allowing diabetic people to profit from best treatments they should receive in order to prevent diabetic complications, by far the main cause of expenses. PMID- 17112162 TI - [Place of radiation-therapy of breast cancer treatment--current and future indications]. AB - Radiotherapy is an important part of breast cancer treatment. After breast conserving surgery, a dose of 50 Grays (Gy) is administered to the entire breast. Boost by external radiotherapy or brachytherapy improves local control especially for women under fifty. For lobular in situ lesions, no additional treatment is required, while intraductal lesions are treated with post-operative radiotherapy in case of Van Nuys score of 7, 8 or 9. After mastectomy, irradiation is proposed in case of skin involvement, invasion of pectoral muscle, positivity of at least 3 axillary lymph nodes, SBR III grade, T3 stage or multifocality where the sum of tumour diameters are >5 cm. Irradiation of the axilla has become exceptional. In node positive patients, the supraclavicular region receives 50 Gy with low energy photons, whereas the internal mammary chain is treated at the same dose but half is administered by low energy photons et half by electrons. Exclusive or preoperative irradiation is rare, because of new chemotherapy schedules, new drugs and hormonotherapy developments. Neoadjuvant chemotherapy enhances possibilities of conservative surgery it is followed by radiotherapy depending on the same indiciations listed above. Re-irradiation is possible but one must be aware of the higher risk of late toxicity. Nowadays, major side-effects are rare but close follow-up during and after treatment is required from the radiation oncologist to detect, evaluate, prevent and even treat possible complications. PMID- 17112163 TI - [Arrhythmogenic right ventricular dysplasia]. AB - Arrhythmogenic right ventricular dysplasia is an unfrequent disease that associates ventricular tachycardia with left bundle branch block morphology and right ventricular fibro-fatty degeneration. The etiology, pathogenesis, criteria for diagnosis and treatment are discussed. PMID- 17112164 TI - [Type 1 diabetes and celiac disease]. AB - Celiac disease is more prevalent in type 1 diabetic patients than in the general population. The exact reason for this association remains unknown. Two hypotheses are taken into consideration: either a common genetic background or an immune response against Langerhans islets triggered by celiac disease. This review presents recent data about this association and its consequences in clinical practice. PMID- 17112165 TI - [DRESS syndrome to sulfasalzine]. AB - We report a case of drug hypersensibility syndrome (DRESS syndrome) developed one month after initiation of a treatment by sulfasalazine. Due to the severity of the cytolytic hepatic damages as observed in this case, we emphasize the importance of suggesting this diagnosis in any patient developing a cutaneous rash and an alteration of the general health status after initiation of a new treatment. The hematologic alterations which represent a key feature for the diagnosis may rash. develop only a few days after the cutaneous rash. PMID- 17112166 TI - [Surgical left ventricular remodelling via endoventriculoplasty: clinical observation]. AB - A case of dilated ischemic cardiomyopathy, with severely impaired left ventricular systolic function, treated by endoventriculoplasty and CABG, is reported. The authors discuss the modern concepts regarding the surgical treatment of large anterior asynergic scars following occlusion of the LAD. Since there are more similarities between akinesia and dyskinesia than previously thought, the endoventriculoplasty of DOR may constitute a new way to surgically remodel the left ventricle in dilated cardiomyopathies with large anterior akinesia. This surgery significantly improves the ventricular function and the overall patients' prognosis at short and mid term follow up. PMID- 17112167 TI - [Vaccination against Rotavirus: an old challenge, an ongoing reality]. AB - Rotavirus are the leading cause of diarrhea and diarrhea related death among infants and young children. Every year rotavirus is associated with over half a million of deathss, mainly in developping countries. Development of a safe vaccine is nowaday the only way to control the disease. A life attenuated oral rotavirus vaccine will be commercialized in a few months in Belgium. PMID- 17112168 TI - [The CHARISMA study: in search of the best antiplatelet strategy for cardiovascular prevention]. AB - The CHARISMA ("Clopidogrel for High Atherothrombotic Risk and Ischemic Stabilization, Management, and Avoidance") trial compared the effects of a dual antiplatelet therapy with clopidogrel plus low dose aspirin with those of a monotherapy with aspirin (75-162 mg/day) on the incidence of cardiovascular events in 15,603 patients at high risk for atherothrombotic events followed for a median of 28 months. The primary efficacy endpoint, a composite of myocardial infarction, stroke, or death from cardiovascular causes, was not significantly different between the two treatment arms. The secondary principal efficacy endpoint, which included all hospitalizations for ischaemic events, was slightly reduced in the group with clopidogrel-aspirin as compared to the group with placebo-aspirin. In a subgroup analysis, among so-called "symptomatico" patients (79 % of the studied population), the dual antiplatelet therapy was associated with a significantly lower incidence of events than aspirin alone, including the primary efficacy end point. On the contrary, in "asymptomatic" patients, such a favourable effect was not observed. Unexpectedly, in this subgroup, a paradoxical increase in the mortality rate was observed with the clopidogrel-aspirin combination. As far as safety was concerned, the risk of severe (difference not significant) and moderate (difference significant) bleeding was higher in patients with the clopidogrel-aspirin combination. In conclusion, acetylsalicylic acid (aspirin) is the first choice drug and the only antiplatelet agent to be used in prvention of cardiovascular disease. In secondary prevention, the addition of clopidogrel may reinforce the cardiovascular protection given by aspirin in "symptomatic" patients, but at the expense of a slightly higher bleeding rate. PMID- 17112169 TI - [The golden years]. PMID- 17112170 TI - [HIV/AIDS. More tests and prevention]. PMID- 17112171 TI - [Housing for the elderly. Living as usual--plus nursing care]. PMID- 17112172 TI - [Assisted living in Urtenen-Schonbuhl. Spitex makes economic living possible]. PMID- 17112173 TI - [Client-centered nursing. Well planned and well controlled]. PMID- 17112174 TI - [Concrete client-centered nursing. Planning and implementing nursing care]. PMID- 17112175 TI - [Gerzensee 2006. Direction markers for nursing career]. PMID- 17112176 TI - [Congress leader Heinz Salzgeber, "Medical technology simplifies work of nurses" (interview by Urs Luthi)]. PMID- 17112177 TI - [A different way]. PMID- 17112178 TI - [Rebecca Spirig, "The key to success is in change" (interview by Margrit Bachl)]. PMID- 17112179 TI - [Timely preparation for change]. PMID- 17112180 TI - [My community of neighbors]. PMID- 17112181 TI - [A specialized service for patients with Alzheimer disease. Architecture for management of patients]. PMID- 17112182 TI - [Transcultural competence. Knowledge, values and stereotypes meeting reality]. PMID- 17112183 TI - [Accompanying perinatal mourning in stillbirth. Death before birth]. PMID- 17112184 TI - [Girls and boys. Mixed in class, different at play]. PMID- 17112185 TI - [Public health. The woman with cats]. PMID- 17112187 TI - Neurosurgeons and their contributions to modern-day athletics: Richard C. Schneider Memorial Lecture. AB - Neurosurgeons in the last half-century have had considerable influence on modern day athletics. In this article, the authors address the contributions made by neurosurgeons as clinician-scientists, particularly as these relate to the understanding and reduction of the incidence and severity of injury to the nervous system during athletic competition. American football has been a proving ground for the ability of the craniospinal axis to withstand and, in unfortunate cases, succumb to tremendous impact forces; in this way, it has served as a model for translational research and was the arena in which Dr. Richard Schneider made his greatest contributions to sports neurosurgery. Therefore, in his memory and in the spirit of the Schneider lectureship, the authors outline the notable contribution to modern-day athletics made by neurosurgeons as it applies to American football. Neurosurgeons have had considerable influence on reducing injury severity, and this cause has been championed by a few notable individuals whose efforts are discussed herein. PMID- 17112188 TI - Subdural hematomas in boxing: the spectrum of consequences. AB - Boxing is a violent sport in which every participant accepts the risk of brain damage or death. This sport has been linked to acute neurological injury and chronic brain damage. The most common life-threatening injury encountered by its participants is subdural hematoma (SDH), and the most feared consequence of chronic insult to the nervous system is dementia pugilistica, or punch drunkenness. Although advances in imaging and neuropsychological testing have improved our ability to diagnose these injuries, the unprecedented sensitivity and wide availability of these modalities have increased the detection of mild cognitive impairment and small, asymptomatic imaging abnormalities. The question has thus been raised as to where on the spectrum of these injuries an athlete should be permanently banned from the sport. In this report the authors describe six boxers who were evaluated for SDH sustained during participation in the sport, and who experienced remarkably different outcomes. Their presentations, clinical courses, and boxing careers are detailed. The athletes ranged in age from 24 to 55 years at the time of injury. Two were female and four were male; half of them were amateurs and half were professionals. Treatments ranged from observation only to decompressive craniectomy. Two of the athletes were allowed to participate in the sport after their injury (one following a lengthy legal battle), with no known sequelae. One boxer died within 48 hours of her injury and at least two suffered permanent neurological deficits. In a third, dementia pugilistica was diagnosed 40 years later, and the man died while institutionalized. PMID- 17112189 TI - Natural antiinflammatory agents for pain relief in athletes. AB - Most athletes experience musculoskeletal injuries during their sports activity that require rest at a minimum, and occasionally injuries are severe enough to necessitate surgical repair. Neurosurgeons are often consulted for athletically sustained injuries and prescribe medications for the associated pain. The use of both over-the-counter and prescription nonsteroidal medications is frequently recommended, but recent safety concerns must now be considered. The authors discuss the biochemical pathways of nonsteroidal drugs and review the potentially serious side effects of these medications. They also review the use of natural supplements, which may be a safer, and often as effective, alternative treatment for pain relief. PMID- 17112190 TI - Concussion in professional football: summary of the research conducted by the National Football League's Committee on Mild Traumatic Brain Injury. AB - PIn 1994 the National Football League (NFL) initiated a comprehensive clinical and biomechanical research study of mild traumatic brain injury (TBI), a study that is ongoing. Data on mild TBIs sustained between 1996 and 2001 were collected and submitted by NFL team physicians and athletic trainers, and these data were analyzed by the NFL's Committee on Mild Traumatic Brain Injury. At the same time, analysis of game videos was performed for on-field mild TBIs to quantify the biomechanics involved and to develop means to improve the understanding of these injuries so that manufacturers could systematically improve and update their head protective equipment. The findings and analysis of the Committee have been presented in a series of articles in Neurosurgery. PMID- 17112191 TI - The role of cadherins and catenins in gliomagenesis. AB - Cell-cell adhesion is a crucial process occurring during normal tissue development. Cadherins are calcium-dependent cell-surface adhesion molecules involved in cell-cell adhesion. They reorganize the actin cytoskeleton via interaction with the catenins. Modulation of the cadherin/catenin system plays a role in cell motility. Dysregulation of the cadherin/catenin assembly has been implicated in various cancers. In this review, the authors summarize all studies focusing on the role of cadherins and catenins in glioma formation. With the emergence of recent data regarding gliomas' putative cell of origin, elucidation of the role of cadherins/catenins in gliomagenesis will become important in devising new therapeutic approaches against such deadly cancers. PMID- 17112192 TI - Reflections on 40 years as a sideline physician. AB - OBJECT: In this study the author presents his personal observations concerning football injuries in a historical perspective with additional literature citations. METHODS: Aspects of brachial plexus and cervical spine injuries, neurapraxia, face mask infractions, concussion, acute subdural hematoma (SDH), and the so-called second-impact syndrome are addressed. CONCLUSIONS: The list of conclusions presented in this paper is as follows: there is more than one kind of brachial plexus injury; wedging of cervical vertebrae may be normal; neurapraxia presents a problem for return to play; face mask injuries are rarely serious; definitions of concussion vary; acute SDH requires immediate transfer to a hospital; and the second-impact syndrome may be a myth to some. PMID- 17112193 TI - Overview of concussion consensus statements since 2000. AB - More refereed publications on sports-related concussion have appeared since 2000 than in all previous years combined. Three international consensus statements, documents from the National Athletic Trainers' Association (NATA) and the American College of Sports Medicine (ACSM), and entire issues of the Clinical Journal of Sport Medicine and the Journal of Athletic Training have been devoted to this subject. The object of this article is to critique the consensus statements and NATA and ACSM documents, pointing out areas of controversy. PMID- 17112194 TI - Lumbar spine injuries in athletes. AB - Lumbar spine injuries in athletes are not uncommon and usually take the form of a mild muscle strain or sprain. More severe injuries sustained by athletes include disc herniations, spondylolistheses, and various types of fracture. The recognition and management of these injuries in athletes involve the additional consideration that to return to play, the lumbar spine must be able to withstand forces similar to those that were injurious. The authors consider common lumbar spine injuries in athletes and discuss management principles for neurosurgeons that are relevant to this population. PMID- 17112195 TI - Return to contact sports after spinal surgery. AB - Decisions regarding the return of injured athletes to contact sports after spinal surgery can be complicated. The authors offer a brief overview of the return-to play guidelines used successfully at their institution for the past two decades when caring for professional and amateur athletes after spinal surgery. PMID- 17112196 TI - Cervical spine injuries in pediatric athletes: mechanisms and management. AB - Sports-related injuries to the spine, although relatively rare compared with head injuries, contribute to significant morbidity and mortality in children. The reported incidence of traumatic cervical spine injury in pediatric athletes varies, and most studies are limited because of the low prevalence of injury. The anatomical and biomechanical differences between the immature spine of pediatric patients and the mature spine of adults that make pediatric patients more susceptible to injury include a greater mobility of the spine due to ligamentous laxity, shallow angulations of facet joints, immature development of neck musculature, and incomplete ossification of the vertebrae. As a result of these differences, 60 to 80% of all pediatric vertebral injuries occur in the cervical region. Understanding pediatric injury biomechanics in the cervical spine is important to the neurosurgeon, because coaches, parents, and athletes who place themselves in positions known to be associated with spinal cord injury (SCI) run a higher risk of such injury and paralysis. The mechanisms of SCI can be broadly subclassified into five types: axial loading, dislocation, lateral bending, rotation, and hyperflexion/hyperextension, although severe injuries often result from a combination of more than one of these subtypes. The aim of this review was to detail the characteristics and management of pediatric cervical spine injury. PMID- 17112197 TI - Back and neck pain in triathletes. AB - OBJECT: As the sport of triathlon has continued to grow, increasing numbers of triathletes have presented in the neurosurgery clinics with various spinal disorders. This epidemiological study was undertaken to establish the lifetime incidence of neck and back pain, to gauge the prevalence of discogenic pain, and to identify risk factors among triathletes in the Boulder, Colorado, area. METHODS: A live online questionnaire was developed that was used to collect information about physical characteristics, training habits, athletic status, number of races completed, and back pain among triathletes. The incidence of cervical and/or lumbar discogenic back pain was defined according to the duration of symptoms for the most recent pain episode. The lifetime incidence of low-back pain was 67.8%, with 23.7% of cases possibly being discogenic in origin. The number of triathlons in which the respondents had participated and the presence of previous sports-related injuries were predictive of low-back pain (p = 0.02 and p < 0.00001, respectively). The lifetime incidence of neck pain was 48.3%, with 21.4% of cases being consistent with intervertebral disc involvement. The number of previous sports-related injuries was predictive of neck pain (p < 0.00001), and a strong tendency toward neck pain was observed for athletes with more total years of participation in sports (p = 0.06). CONCLUSIONS: The two main risk factors for long-term spinal problems include sports-related injuries and overuse. The study results definitely support the influence of both mechanisms for low-back pain. Neck pain was associated with an injury event, and a strong (although not statistically significant) tendency toward neck pain was observed in respondents with overuse injuries. PMID- 17112198 TI - Sideline and ringside evaluation for brain and spinal injuries. AB - Participation in contact and collision sports carries an inherent risk of injury to the athlete, with damage to the nervous system producing the most potential for significant morbidity and death. Neurological injuries suffered during athletic competition must be treated promptly and correctly to optimize outcome, and differentiation between minor and serious damage is the foundation of sideline/ringside management of the injury. In this article the authors present a guide to the sideline or ringside identification and management of head and spinal injuries. PMID- 17112199 TI - Participation in contact or collision sports in athletes with epilepsy, genetic risk factors, structural brain lesions, or history of craniotomy. AB - Despite a plethora of guidelines for return to play following mild head injury, a discussion of when and if an athlete should be allowed to participate in contact or collision sports if he or she sustains a structural brain lesion or after a head injury requiring craniotomy is lacking. The structural lesions discussed include arachnoid cyst, Chiari malformation Type I, cavum septum pellucidum, and the presence of ventriculoperitoneal shunts. Issues unique to this population with respect to the possibility of increased risk of head injury are addressed. The population of athletes with epilepsy and certain genetic risk factors is also discussed. Finally, the ability of athletes to participate in contact or collision sports after undergoing craniotomies for traumatic or congenital abnormalities is evaluated. Several known instances of athletes returning to contact sports following craniotomy are also reviewed. PMID- 17112200 TI - Carotid endarterectomy: current indications for surgery. AB - The study of carotid artery occlusive disease interventions can be divided clinically into the treatment of asymptomatic and symptomatic diseases. Clinical trials that have studied or are currently studying asymptomatic disease include: the Carotid Artery Stenosis with Asymptomatic Narrowing Operation Versus Aspirin study; the Mayo Asymptomatic Carotid Endarterectomy trial; Veterans Administration Cooperative Trial on Asymptomatic Carotid Stenosis; and the Asymptomatic Carotid Atherosclerosis Study. Trials for the treatment of symptomatic disease include: the North American Symptomatic Carotid Endarterectomy Trial; the European Carotid Surgery Trial; and the Veterans Administration Cooperative Study. In the earliest trials conducted to study asymptomatic disease medical therapy was slightly favored; on close scrutiny these studies were flawed and the findings appeared to be equivocal. The more scientific and appropriate trial, which was ended due to ethical concerns, revealed a clear advantage in patients who underwent surgery for greater than 60% stenosis and when the surgical center demonstrated less than 3% surgical risks. All trials studying symptomatic disease found a significant decrease in subsequent stroke when surgical intervention was performed. It is now judged that patients with greater than 50% stenosis receive significant benefit. In this paper the authors review the data from all of these studies. They also review data for special circumstances, such as critical stenosis and patients with symptomatic and asymptomatic Hollenhorst plaques. It is their opinion that these data have allowed surgeons to make much more educated decisions when considering the treatment of patients with carotid artery occlusive disease. PMID- 17112201 TI - Carotid endarterectomy compared with angioplasty and stenting: the status of the debate. AB - Endarterectomy is the treatment of choice for patients with symptomatic stenosis of the internal carotid artery. Recently, debate has arisen over the potential benefits of endovascular techniques. Although retrospective analyses of angioplasty and stenting procedures suggest comparable clinical efficacy to endarterectomy, prospective evaluation is pending. The authors review the status of the debate and discuss those issues on both sides that are particularly contentious and clinically relevant. PMID- 17112202 TI - Angioplasty and stenting for carotid artery stenosis: indications, techniques, results, and complications. AB - Following the favorable results obtained in the treatment of coronary artery disease, combined angioplasty and stenting has been advocated for the treatment of carotid artery stenosis as well. Although widespread application of angioplasty and stenting for carotid artery disease is neither indicated nor recommended, it may be a viable alternative therapy for select patients who are high-risk patients for surgery. The results of early series have suggested that endoluminal revascularization in these high-risk patients can be performed with an acceptable degree of safety. Although the incidence of death and major stroke rates following angioplasty and stenting procedures compares favorably with surgery, results of more recent clinical series have suggested that the incidence of perioperative transient neurological events and minor strokes may be higher than suggested by earlier reports, especially in patients with recent neurological symptoms and "unstable" plaques. In this article, the authors review the current potential indications for and preliminary results of angioplasty and stenting and describe their procedural technique. In addition, potential applications of stenting to intracranial thromboocclusive carotid artery disease are reviewed. PMID- 17112203 TI - Complications of carotid angioplasty and stenting. AB - The authors report the complications that occurred in their experience with performing recanalization procedures in the internal carotid artery and present their treatment strategies. The complications can be classified into those that were periprocedural and those that were postprocedural. The former include complications related to the vascular-approach access site of and those associated with the dilation and stenting procedure. Other complications observed included embolic events, dissection, vascular spasm, bradycardia, inappropriate dilation, occlusion of the external carotid artery, and rare, unusual complications such as the occurrence of iatrogenic cavernous carotid fistula. Postprocedure complications occurred in the hours and days following the procedure in the form of embolic and occlusive events, and hypotension and bradycardia were seen as late complications in the months following the procedure. The authors discuss how such complications occur and provide suggestions on how to avoid them. The role of stent placement and the potential use of protective devices are explored. Overall, adequate use of currently available systems allows for safe application of endovascular treatment techniques that avoid altogether or treat these potential complications. A reduced incidence of complications related to the initial individual learning curve may be obtained with preclinical training, in which use of invitro models should be considered. Surgical standby no longer seems required; however, early posttreatment surveillance in intensive care unit is mandatory to avoid the remaining primary complications. PMID- 17112204 TI - Moyamoya disease in the midwestern United States. AB - There have been few investigations of moyamoya disease in the United States and no systematic description of the management practices or outcome from this population. The authors reviewed their experience with this disease to gain a better understanding and improve the treatment of patients with moyamoya disease in the United States. Over a 25-year period 30 patients with moyamoya disease have been treated at the University of Iowa. The cases were divided into patients who had classic, probable, and akin moyamoya disease. Results indicated that there was a bimodal age distribution and a female predominance of cases. In estimating the referral pattern of our institution, the authors determined that there were greater numbers of epidemiological characteristics than previously anticipated. Patients were treated either surgically or nonsurgically, and different management strategies were utilized in each of the major groups: superficial temporal artery to middle cerebral artery anastomosis and encephalodurosynangiosis in the surgical group; or antiplatelet, anticoagulation, or nonpharmacological intervention in the nonsurgical group. The authors conclude that there is a higher prevalence and incidence of moyamoya disease in the United States than previously reported and that there are some clinical characteristics of this disease that differ from the cases reported in southeast Asia. These differences may be due to genetic or environmental factors but can also be partly explained by the lower index of suspicion for this disease and, thus, a delay in or complete absence of the correct diagnosis. PMID- 17112205 TI - Role of basic fibroblast growth factor in the pathogenesis of moyamoya disease. AB - The pathogenesis of moyamoya disease is still under investigation. In this study, the authors focus on the role of cytokines in the pathogenesis of moyamoya disease by using immunohistochemical analyses. The authors examined two specimens in the circle of Willis obtained at autopsy from two patients with moyamoya disease and two additional specimens obtained from control cadavers with atherosclerotic stenosis of the intracranial carotid arteries. Immunohistochemical examinations of the sections of the major intracranial arteries were performed using antismooth muscle cells (SMCs), monocytes, growth factor, cell nuclear antigen, and fragmented DNA antibodies. Basic fibroblast growth factor (bFGF) staining was present only in the endothelial cells of the moyamoya disease specimens and was not seen in control samples. In addition, the endothelial cells and SMCs in the media were positive for terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end labeling of fragmented DNA method but not in the SMCs in the intima in moyamoya disease specimens, which indicates that an apoptotic process is active in only SMCs in the media but not in the intima. In conclusion, it is suggseted that the presence of bFGF in the media specifically seen in moyamoya disease suppresses the apoptotic process of SMCs in the intima. PMID- 17112206 TI - Comparison of positron emission tomography study results of cerebral hemodynamics in patients with bleeding- and ischemic-type moyamoya disease. AB - The purpose of this study was to elucidate the difference in cerebral hemodynamics and metabolic status between patients with bleeding- and ischemic type moyamoya disease. Regional cerebral blood flow (rCBF), regional cerebral metabolic rate of oxygen (rCMRO2), regional oxygen extraction fraction (rOEF), and regional cerebral blood volume (rCBV) in the cortex of the middle cerebral artery (MCA) territories and rCBV in the striatum were measured using positron emission tomography (PET) in 17 patients with moyamoya disease. Patients were divided into three subgroups according to type of disease manifestation and age: adult bleeding type (five cases), adult ischemic type (10 cases), and childhood ischemic type (two cases). When compared with adult controls, statistically significant reductions in rCBF and rCMRO2, elevation in rOEF in the MCA territories, and elevation of rCBV in the striatum were observed in PET studies for all three subgroups. Between the adult bleeding type and ischemic type, rCBF, rCMRO2, and rOEF in the MCA territories were not different, but rCBV in the striatum was higher in patients with ischemic-type moyamoya disease than in those with the bleeding type. In adult patients with bleeding and ischemic types, rOEF and rCBV in the MCA territories and rCBV in the striatum were significantly lower than in patients with childhood ischemic-type moyamoya disease. In adult patients with bleeding-type moyamoya disease, cerebral hemodynamics were impaired and similar to those in adult ischemic type. PMID- 17112207 TI - Multiple combined indirect procedure for the surgical treatment of children with moyamoya disease. A comparison with single indirect anastomosis and direct anastomosis. AB - Considering three different bypass procedures now in use, (single indirect nonanastomotic bypass procedure, multiple combined indirect (MCI) nonanastomotic procedure and direct anastomosis), the authors attempted to identify the most appropriate bypass procedure for treating ischemic-type moyamoya disease in children. The authors performed three procedures (the original encephaloduroarteriosynangiosis [EDAS] alone, the frontotemporoparietal combined indirect bypass procedure, and the superficial temporal artery--middle cerebral artery [STA-MCA] anastomosis with encephalomyosynangiosis [EMS]) on 72 hemispheres in 50 patients with pediatric moyamoya disease. Analyses were then performed to compare postoperative collateral vessel formation found on angiograms, complications, and clinical improvements. Postoperative collateral formations were observed in more than two-thirds of the MCA distribution after the EDAS alone, the MCI procedure, and the direct anastomosis in 44%, 52%, and 74% of the surgically treated hemispheres, respectively. In addition, frontal encephalomyoarteriosynangiosis of the MCI bypass procedure formed collateral vessels of the anterior cerebral artery distribution in 94% of the treated hemispheres. Postoperatively, clinical symptoms resolved in 56%, 63%, and 74% of the treated sides 1 year after EDAS alone, MCI procedure, and the direct anastomosis, respectively. One patient suffered a minor stroke after EDAS alone, two patients developed epidural hematomas after the MCI procedure, and one patient suffered a major stroke and one patient a minor stroke after undergoing direct anastomosis. The direct anastomosis procedure was found to result in the best postoperative collateral vessel formation and clinical improvement. However, the single and multiple combined indirect nonanastomotic bypass procedures were found to be safer than direct anastomosis. Furthermore, the frontotemporoparietal combined indirect bypass procedure caused the formation of collateral circulation not only in the MCA but also in the ACA distribution. Based on analysis of these findings, the authors recommend the MCI procedure as the appropriate surgical procedure in the treatment of children with moyamoya disease, although the best treatment is the STA-MCA anastomosis with EMS when feasible. PMID- 17112208 TI - Long-term outcome after STA-MCA anastomosis for moyamoya disease. AB - A long-term assessment was performed to determine the posttreatment clinical course of 113 patients with moyamoya disease. All patients sustained cerebral ischemic attacks and underwent superficial temporal artery-middle cerebral artery anastomosis with or without temporal muscle grafting. The follow-up duration was 3 to 24 years (mean 14.4 +/- 5.8 [standard deviation]). Complete cessation of the ischemic episodes was obtained in 110 of 113 patients. One hundred patients were able to return to independent acitvities of daily living. Intellectual delays prevented 24 patients from engaging in an independent social life . Although intracranial bleeding is one of the common manifestations in moyamoya disease, hemorrhage was not detected in the 113 patients who underwent cerebral revascularization. PMID- 17112209 TI - Arteriovenous malformation in association with moyamoya disease. Report of two cases. AB - The authors report two rare cases of arteriovenous malformation (AVM) associated with moyamoya disease. An AVM, supplied by transdural communicating arteries, was located in the right occipital lobe in one patient who presented with ischemia. The second AVM, which was supplied by basal moyamoya vessels, was located in the posterior part of the left frontal lobe in a patient who developed intracerebral hemorrhage that occupied the left basal ganglion. A review of the literature revealed a total of 12 AVMs in 11 patients with moyamoya disease including our cases. All AVMs were cerebral and two were supplied by normal cerebral arteries, whereas six AVMs were supplied by basal moyamoya vessels at the base of the brain and four AVMs were supplied by external carotid arteries through the transdural communicating arteries. Every AVM drained into deep or cortical cerebral veins. These findings suggest that the hyperangiogenic character of moyamoya disease occasionally induces the development of acquired arteriovenous shunts that mimic AVM. PMID- 17112210 TI - Moyamoya syndrome associated with cocaine abuse. Case report. AB - The authors report the case of a 30-year-old woman who was a long-term intranasal cocaine abuser and who presented with transient ischemic attacks and multiple cerebral infarctions that were associated with moyamoya syndrome. The authors suggest that, because of its sympathomimetic effects, chronic cocaine use may promote intracranial arterial stenosis, distal ischemia, and subsequent formation of moyamoya-like vessels. The patient has remained clinically stable with no new episodes of stroke 6 years after undergoing "pial synangiosis" (modified encephaloduroarteriosynangiosis) to revascularize both hemispheres. Cocaine abuse may lead to moyamoya syndrome and may represent a chronic effect on the cerebral vasculature. PMID- 17112211 TI - Current concepts on carotid artery-cavernous sinus fistulas. AB - With greater understanding of the pathophysiological mechanisms by which carotid artery-cavernous sinus fistulas occur, and with improved endovascular devices, more appropriate and definitive treatments are being performed. The authors define cartoid cavernous fistulas based on an accepted classification system and the signs and symptoms related to these fistulas are described. Angiographic evaluation of the risk the lesion may pose for precipitating stroke or visual loss in the patient is discussed. The literature on treatment alternatives for the different types of fistulas including transvenous, transarterial, and conservative management is reviewed. PMID- 17112212 TI - Percutaneous transluminal angioplasty of intracranial artery stenosis: clinical results in 24 patients. AB - Circumscribed stenotic lesions of the intracranial arteries can cause cerebral ischemia by hemodynamic and/or thromboembolic mechanisms. Anticoagulation therapy, antiplatelet therapy, and bypass surgery are treatment strategies that have no direct impact on the underlying lesion. This study summarizes the experience of a single institution at which percutaneous transluminal angioplasty (PTA) of intracranial atherosclerotic stenoses was performed. The authors performed a retrospective analysis of 24 consecutive patients. Their medical histories (cardiovascular risk factors, current clinical signs and symptoms and their duration, previous stroke[s], and medical treatment) were evaluated together with findings from previous imaging studies. The site and degree of the stenoses to be treated (target lesion) were identified with the use of ultrasound and angiography studies. Additional vascular stenoses were noted. Percutaneous transluminal angioplasty was performed using single-lumen balloon microcatheters with appropriate diameters. The results of PTA were correlated with angiographic and ultrasound findings and the clinical outcome. Significant cardiovascular risk factors and clinical signs and symptoms related to the target lesion that persisted despite medical treatment were identified in all patients except one. The duration of symptoms varied from several days to 8 months. Previous stroke had occurred in four patients. The degree of stenosis was classified as "high grade" in 10 patients and as "subtotal" in 14. The target lesion (stenosis) was located in the anterior circulation in eight patients (four in the internal carotid and four in the middle cerebral arteries). Stenoses of posterior circulation vessels were treated in 16 patients (nine vertebral, six basilar, and one posterior cerebral arteries). Recanalization was rated "complete" in 15 patients and sufficient in six patients. In three patients residual stenosis remained. Complications were encountered in seven patients: two asymptomatic dissections, one transient vessel occlusion, one vessel occlusion with subsequent stroke, and three ischemic lesions likely due to thromboembolism, two of which caused only transient neurological symptoms. Percutaneous balloon dilation proved effective in the treatment of intracranial atherosclerotic stenosis. There are, however, potential complications and experience with this procedure is only limited. Long-term results need to be determined. The authors conclude from their preliminary results that PTA may be an alternative to bypass surgery and conservative management and may be considered for patients in whom ischemic neurological symptoms persist despite medical treatment. PMID- 17112213 TI - Carotid angioplasty and stenting for recurrent and radiation-induced stenosis: preliminary experience. AB - Carotid endarterectomy for atherosclerotic occlusive disease has become the standard of care for the treatment of symptomatic and asymptomatic occlusive disease of the carotid bifurcation, based on the results of the North American Symptomatic Carotid Endarterectomy Trial, as well as the Asymptomatic Carotid Atherosclerosis Study. For surgical treatment to be of benefit, the perioperative complication rate for neurological events should be 6% or less in the symptomatic population and 3% or less in the asymptomatic group. The performance of carotid endarterectomy for recurrent stenosis and radiation-induced stenosis has reported neurological events ranging from 4 to 10%. It is in this particular population that carotid angioplasty and stent placement may play a role. The authors performed a retrospective analysis of 11 patients who underwent carotid angioplasty and stent placement for recurrent or radiation-induced stenosis. One patient in whom endarterectomy was performed by the vascular surgery service had a critical stenosis distal to the endarterectomy site and awoke with a neurological deficit. This patient underwent reexploration and placement of a stent in the artery distal to the arteriotomy site. The follow-up period ranged from 7 to 12 months. Patient age ranged from 65 to 77 years (mean 75 years). Five of eight patients underwent angioplasty and stent placement for recurrent atherosclerotic disease. Two patients had radiation-induced stenosis, and one patient had a stent placed intraoperatively. All patients, with the exception of the one who underwent intraoperative stent placement, had posttreatment stenoses of less than 15%. The surgical patient had a 30% residual stenosis distally. There were no intra- or postoperative transient ischemic attacks, major or minor strokes, or deaths. Patients who have recurrent or radiation-induced stenosis are potential candidates for angioplasty and stent placement. Before this can be recommended as an alternative to surgical correction, a longer follow-up period is required. PMID- 17112214 TI - Stent placement for vertebral artery occlusive disease: preliminary clinical experience. AB - Symptoms of vertebrobasilar insufficiency may precede neurological sequelae in up to 50% of patients. Although select patients may benefit from microsurgical revascularization, combined perioperative morbidity and mortality rates can be as high as 20%. The authors present their preliminary clinical experience using stent placement for symptomatic vertebral artery (VA) occlusive disease. Six patients with clinical symptoms of vertebrobasilar insufficiency in whom VA stents were placed from 1995 to 1998 were identified. Diagnostic four-vessel cerebral angiography identified causative stenotic, atherosclerotic lesions in all cases. A transfemoral or transradial artery approach after the patient had undergone full heparinization was chosen for endovascular stenting. Guidewire placement across the lesion followed by urokinase infusion preceded stenting. Prestent angioplasty was performed in two patients. Following the procedure, all patients were maintained on daily antiplatelet therapy. Patient age ranged from 45 to 76 years (average 63 years). Four patients were men and two were women. Angiography revealed greater than 95% stenosis in five patients and greater than 70% stenosis in one. Three patients had complete occlusion of the contralateral VA; in one other, the VA supplied only the posterior inferior cerebellar artery; and the remaining two patients had VAs with greater than 70% stenosis. Ten stents were placed in six patients for five VA origin lesions and one distal VA stenosis. A VA dissection occurring poststenting was treated by placement of three additional stents. One patient had transient double vision. All had resolution of their presenting symptoms. Follow up ranged from 1 to 24 months (average 8.4 months). Angiograms obtained in four patients at least 3 months postprocedure have revealed stent patency in all cases without evidence of restenosis. Vertebral artery stent placement can be safely performed and is a viable treatment option for carefully selected patients with vertebrobasilar insufficiency. PMID- 17112215 TI - Therapeutic options for endovascular therapy for intracranial aneurysms. AB - Endovascular surgical technology is in the early stages of evolution. A critical phase of this development has been microcatheter technology, which has permitted sufficiently precise intravascular navigation to safely engage the lumen of the aneurysm itself. Digital subtraction angiography, rapid filming techniques and image acquisition, and simultaneous multiplanar imaging capability are indispensable tools that are constantly being refined in the setting of ever improving computer technology. The marriage of these different technologies has allowed effective endovascular treatment of difficult-to-access aneurysms in medically compromised patients for whom open microsurgery has inherently higher risks. PMID- 17112216 TI - Safety and efficacy of transluminal balloon angioplasty in the prevention of vasospasm in patients with Fisher Grade 3 subarachnoid hemorrhage: a pilot study. AB - Recent advances in neuroradiology have made it possible to dilate human cerebral arteries that show vasospasm following aneurysmal subarachnoid hemorrhage (SAH), but the time window is short and the success rate for reversal of delayed ischemic neurological deficit (DIND) varies between 31% and 77%. In a canine model of vasospasm, transluminal balloon angioplasty (TBA) performed on Day 0 (the day of aneurysm rupture) has been shown to completely prevent the development of angiographically demonstrated narrowing by Day 7; this effect is better than any pharmacological treatment for vasospasm thus far described. The authors conducted a pilot trial to assess the safety and efficacy of TBA performed within 3 days post-SAH. Twelve patients with a very high probability of developing vasospasm (Fisher Grade 3) were included. Target vessels for prophylactic TBA were the internal carotid artery, A1 segment, M1 segment, and P1 segment bilaterally, the basilar artery, and the vertebral artery. No patient developed DIND or more than mild vasospasm, according to transcranial Doppler criteria. At 3 months, seven patients made a good recovery, two patients were moderately disabled, and three patients died; one patient died because of a vessel rupture during TBA and two older patients died of medical complications associated with an already poor clinical condition at admission. Compared with the results of large series reported in literature of patients with aneurysmal SAH, the results of this pilot study indicate an extremely low incidence of vasospasm and DIND after patients underwent prophylactic TBA. A larger, randomized study, however, is required to determine whether prophylactic TBA is efficacious enough to justify the risks. PMID- 17112217 TI - Endovascular treatment for symptomatic cerebral vasospasm after subarachnoid hemorrhage: transluminal balloon angioplasty compared with intraarterial papaverine. AB - The authors retrospectively evaluated the short-term neurological improvement of 69 patients undergoing endovascular treatment for symptomatic cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH). The patient group observed here is a subset of patients enrolled in the multicenter North American Trial of Tirilazad in SAH. Thirty-one patients were treated with intraarterial administration of papaverine (IAP). Fourteen patients were only treated with transluminal balloon angioplasty (TBA), and 24 patients received a combination of angioplasty and papaverine. The purpose of this study was to compare the effects of IAP and TBA on short-term clinical improvement of patients. Daily clinical staging with the modified Glasgow Coma Scale and every-other-day transcranial Doppler (TCD) measurements allowed for a close investigation of the clinical course. Furthermore, this study was designed to investigate the effects of treatment timing on short-term outcome. Although TCD studies demonstrated a decrease in flow velocities in the middle cerebral artery in both treatment groups, indicating a vasodilating effect of both treatment modalities (dv = -18.4 cm/second for papaverine, dv = -26.04 cm/second for angioplasty; p = 0.5509), there was no significant difference in clinical improvement at Days 1 and 4 postprocedure (p = 0.1996). Neither of the two treatment forms showed an effect of therapy timing on neurological outcome. Neither IAP nor TBA was correlated with a high percentage of short-term neurological improvement. The authors discuss reasons why those procedures may result in limited clinical change. PMID- 17112218 TI - Early and aggressive treatment of medically intractable cerebral vasospasm with pentobarbital coma, cerebral angioplasty and ICP reduction. AB - The authors present the unique experience of one neurovascular service under the direct supervision of the senior author, for which surgical, endovascular, and intensive care treatments were conducted in a select group of 32 patients with aneurysmal subarachnoid hemorrhage who had medically intractable symptomatic vasospasm. A protocol of early and aggressive treatment was instituted using pentobarbital coma, cerebral angioplasty, and intracranial pressure (ICP) reduction. The patient population consisted of 25 women and seven men, whose ages ranged from 34 to 60 years (average 47 years). The patients' Hunt and Hess grades on presentation were as follows: Grade 0 (one); Grade I (three); Grade II (two); Grade III (nine); Grade IV (10); Grade V (seven). Microsurgical clipping alone was performed in 15 of 32 patients, endosaccular occlusion was performed in 17 of 20 patients, and two patients underwent combined treatment. Subsequent angioplasty was performed in 26 of 32 patients. Additionally, all 32 patients underwent treatment of increased ICP with ventriculostomy placement, removal of the bone flap (11), evacuation of associated intracranial hematoma (five), and decompressive obectomy (four). Twenty-one patients survived and 11 died. Of the 21 survivors, seven have returned to work, live independently, and have no neurological deficits; eight require minimal assistance at home; four are in rehabilitation with moderate deficits at 3 months; and two remain in a persistent vegetative state. In this group of aggressively treated patients who received pentobarbital cerebral protection, successful treatment of medically intractable cerebral vasospasm was related to time of treatment (< 2 hours), expeditious reduction of elevated ICP, and angioplasty. PMID- 17112219 TI - Outcomes after gamma knife radiosurgery in solitary acoustic tumors and neurofibromatosis Type 2. AB - Surgeons perform stereotactic radiosurgery as the main alternative to acoustic tumor (vestibular schwannoma) resection. The goals of radiosurgery include preservation of neurological function and prevention of tumor growth. Longer-term outcomes are not well documented for patients with solitary tumors or those with neurofibromatosis Type 2 (NF2). To define outcomes, the authors evaluated 462 consecutive patients with solitary acoustic tumors and 40 patients with NF2 (total of 45 tumors treated) who underwent radiosurgery between 1987 and 1998. Serial imaging studies, clinical evaluations, and a patient survey were performed. The average tumor margin dose was 15 Gy, and the mean transverse tumor diameter was 22 mm. In patients with solitary tumors, prior resection had been performed in 111 patients (24%); 27 patients experienced tumor recurrence after a "total resection." The clinical tumor control rate (no resection required) was 98%. In non-NF2 patients followed for at least 5 years, 100 tumors (61.7%) were smaller, 53 (32.7%) remained unchanged in size, and nine (5.6%) were slightly larger. Resection was performed in four patients (2.4%). Neurological deficits after radiosurgery all occurred within the first 28 months. The rates of facial and trigeminal neuropathy varied with radiosurgery technique. In patients with NF2, 16 tumors were smaller, 28 remained unchanged, and one enlarged (overall 98% control rate at median 3-year follow up). Resection was performed in three patients (7%). Useful hearing was preserved in six (43%) of 14 NF2 patients who had useful hearing before radiosurgery. Radiosurgery provided long-term tumor control associated with high rates of neurological function preservation. No further tumor surgery was necessary in 98% of patients with solitary tumors followed for a minimum of 5 years. PMID- 17112220 TI - Quality of life following acoustic neuroma surgery. AB - In the treatment of acoustic neuroma, operative results have improved greatly during recent years, with high rates of functional cranial nerve preservation. Because of this, it has become more important to consider issues of patient satisfaction and quality of life (QOL) following treatment for these lesions. The authors have developed a novel questionnaire designed to measure QOL in patients with acoustic neuromas, and they administered it to 50 consecutive patients at least 6 months after acoustic neuroma surgery. Overall QOL was judged to be good but with definite minor difficulties, including some problems with hearing, facial nerve function, headache, tinnitus, dizziness, activity level, enjoyment of life, and emotional well-being. No significant differences were found between age groups and different operative approaches, and only minor differences were found in relation to tumor size. Patients with intracanalicular tumors fared no better than those with cerebellopontine angle tumors. Analysis of the data suggests an overall good outcome from acoustic neuroma surgery; however, when discussing the possible effects on postoperative QOL, even the potential minor problems should not be minimized, especially in patients undergoing operation for small or intracanalicular tumors. PMID- 17112221 TI - Late-onset facial nerve degeneration after vestibular schwannoma surgery: incidence, putative mechanisms, and prevention. AB - Delayed facial nerve dysfunction after vestibular schwannoma surgery is a poorly understood phenomenon that has been reported to occur in 15 to 29% of patients undergoing microsurgery. It is a condition characterized by spontaneous deterioration of facial nerve function in a patient who has otherwise normal or near-normal facial function in the immediate postoperative period. This delayed paralysis is generally reported to occur in the first few days postsurgery, with the majority of patients eventually recovering their immediate postoperative facial function. However, infrequently, it can also occur more than 1 week after surgery (so-called late-onset facial nerve palsy). The authors reviewed facial nerve outcome in 611 patients who underwent microsurgery between 1973 and 1994. The facial nerve was anatomically preserved in 596 patients (97.5%), and 90% of patients had House-Brackmann[6] Grade 1 or 2 function 1 year after surgery. Late onset facial dysfunction was seen in 13 patients (2.1%). All of these had significant deterioration in facial nerve function between 1 and 4 weeks postoperatively, and all showed improvement by 1 year. In this study, the focus on these patients who developed late-onset facial palsy. The incidence, treatment strategies, and outcomes will be discussed with emphasis on possible pathophysiological mechanisms that contribute to this relatively rare condition. PMID- 17112222 TI - Decompression of the labyrinthine segment of the facial nerve in acoustic neuroma surgery: a consideration for minimizing postoperative delayed facial nerve dysfunction. AB - Delayed facial nerve palsy, a condition characterized by spontaneous deterioration of facial nerve function in patients who had otherwise normal or near-normal facial function in the immediate postoperative period, has been reported in 15 to 29% of patients undergoing microsurgical resection of vestibular schwannomas. One putative mechanism for its occurrence suggests that edematous entrapment of the facial nerve in the meatal foramen (the narrowest segment of the internal auditory canal) may lead to nerve ischemia or necrosis and subsequent facial nerve dysfunction. To assess whether meatal decompression may help reduce the incidence of delayed facial nerve palsy during microsurgical resection of acoustic tumors, we compared 25 patients undergoing translabyrinthine removal of acoustic neuromas who received prophylactic decompression of the labyrinthine segment of the facial nerve (Group 1) with 40 patients who did not receive facial nerve decompression (Group 2). No patients in Group 1 had a delayed progressive facial paralysis with degeneration. In contrast, when Group 2 patients with larger, average-sized tumors were reviewed, eight patients (20%) developed delayed degeneration. These findings suggest that decompression of the labyrinthine segment may be of value in acoustic tumor surgery in reducing delayed facial nerve dysfunction. Further study is indicated in this important area. PMID- 17112223 TI - Endoscopic anatomy of the cerebellopontine angle: a study in cadaver brains. AB - The increasing trend toward performing minimally invasive neurosurgery may benefit from recent progress in using neuroendoscopic techniques to reduce trauma in patients who have undergone operations. Arterial and venous vessels, especially loops, may compress the central segment and cause hyperactive dysfunction of the nerves. Relationships of the anterior inferior cerebellar artery to the facial and vestibulocochlear nerves and the anterior inferior, and superior cerebellar arteries to the trigeminal nerve were studied. The authors report findings from an endoscopic study performed in cadaver heads via the retrosigmoid and retrolabyrinthine approaches. Arteries and veins were colored by injection of red and blue silicon rubber. The cerebellopontine angle (CPA) was examined using 2.7-mm and 4-mm-diameter rigid endoscopes at viewing angles of 0s degrees , 30s degrees , and 70s degrees . Well-known structures could be identified endoscopically without prior dissection, and the entire CPA could be explored. However, with a retrosigmoid or a retrolabyrinthine approach, the cerebellum had to be retracted to some extent to view the CPA. Moreover, wide dural exposure was required to maneuver the endoscope freely in the CPA. Use of the rigid fiberoptic endoscope is not yet superior to standard surgical techniques for approaching and exploring the CPA. PMID- 17112224 TI - Hemorrhagic vestibular schwannoma: an unusual clinical entity. Case report. AB - Hemorrhagic vestibular schwannomas are rare entities, with only a few case reports in the literature during the last 25 years. The authors review the literature on vestibular schwannoma hemorrhage and the presenting symptoms of this entity, which include headache, nausea, vomiting, sudden cranial nerve dysfunction, and ataxia. A very unusual case is presented of a 36-year-old man, who unlike most of the patients reported in the literature, had clinically silent vestibular schwannoma hemorrhage. The authors also discuss the management issues involved in more than 1000 vestibular schwannomas treated at their institution during a 25-year period. PMID- 17112225 TI - Future of toxicology--mechanisms of toxicity and drug safety: where do we go from here? AB - Recent high-profile drug withdrawals increase the pressure on regulators and the pharmaceutical industry to improve preclinical safety testing. Understanding mechanisms of drug toxicity is an essential step toward improving drug safety testing by providing the basis for mechanism-based risk assessments. Nonetheless, despite several decades of research on mechanisms of drug-induced toxicity and the application of various new technologies to preclinical safety assessment, the overall impact on preclinical safety testing has been modest. Assessing the risk of exposing humans to new drug candidates still depends on preclinical testing in animals, which in many, but not all cases, predicts outcomes in humans accurately. The following offers a perspective on the challenges and opportunities facing efforts to improve preclinical safety testing and outlines gaps and needs that must be addressed. A case is built for focusing solutions on defined problems within the current safety testing paradigm rather than imposing wholesale change. Targets for application of new technologies, including in silico screening, biomarkers, surrogate assays and 'omic technologies, are outlined. Improving drug safety testing will depend on improving the application of mechanism-based risk assessment but will also require improving public and private collaborations in order to focus research regarding the mechanism of drug induced toxicity on the most important problems. PMID- 17112226 TI - A review of the role of the sequence-dependent electrostatic landscape in DNA alkylation patterns. AB - Alkylating agents, including environmental and endogenous carcinogens and DNA targeting antineoplastic agents, that adduct DNA via intermediates with significant cationic charge show a sequence selectively in their covalent bonding to nucleobases. The resulting patterns of alkylation eventually contribute to the agent-dependent distributions and types of mutations. The origin of the regioselective modification of DNA by electrophiles has been attributed to steric and/or electronic factors, but attempts to mechanistically model and predict alkylation patterns have had limited success. In this review, we present data consistent with the role of the intrinsic sequence-dependent electrostatic landscape (SDEL) in DNA that modulates the equilibrium binding of cations and the bonding of reactive charged alkylating agents to atoms that line the floor of the major groove of DNA. PMID- 17112227 TI - cis-Terpenones as an effective chemopreventive agent against aflatoxin B1-induced cytotoxicity and TCDD-induced P450 1A/B activity in HepG2 cells. AB - Aflatoxin B1 (AFB1) is a potent carcinogen, which can significantly increase the risk of hepatocellular carcinoma development through food contamination. In past decades, chemopreventive agents, such as oltipraz and chlorophyllins, have demonstrated that chemo-intervention is an effective approach to reduce hepatotoxicity by AFB1. However, because of the potential adverse effects of these agents, alternative novel mechanism-based chemopreventive agents are needed. We report here that novel cis-terpenones 1-3, which were synthesized as the precursors of natural product analogues in our laboratory, showed promising protective effects against AFB1-induced cytotoxicity in HepG2 cells. Chemo protection was observed with increasing concentrations of cis-terpenones in the co-treatment of AFB1, and no cytotoxicity was observed with cis-terpenones alone. In addition, cis-terpenones 1-3 at 10 and microM effectively inhibited induced cytochrome P450 1A/1B activity by 50% in HepG2 cells, as indicated by an EROD assay. P450 1A/B is involved in the activation of many pre-carcinogens and is highly inducible in liver cells. These results suggested that novel terpenones 1 3 are candidates for the development of novel mechanism-based chemopreventive agents against AFB1 and other carcinogenic stimuli. PMID- 17112228 TI - Hydroxylated polychlorinated biphenyls are substrates and inhibitors of human hydroxysteroid sulfotransferase SULT2A1. AB - Polychlorinated biphenyls (PCBs) are important persistent environmental contaminants. PCBs can be metabolically converted to their hydroxylated metabolites (OHPCBs), and in recent years, these OHPCBs have been observed to inhibit human sulfotransferases (SULTs) such as the phenol SULTs (SULT family-1) involved in the metabolism of estrogen and various other endogenous and xenobiotic phenols. In the present study, we have investigated the hypothesis that OHPCBs interact with family 2 hydroxysteroid (alcohol) SULTs (e.g., human SULT2A1), enzymes that are physiologically important for the metabolic transformations of several key endogenous hydroxysteroids as well as xenobiotic alcohols. We have examined the interactions of three OHPCBs with purified recombinant human SULT2A1 (also known as either human DHEA-ST or ST2A3). These studies with SULT2A1 were carried out on 4'-hydroxy-2,5-dichlorobiphenyl (4'-OH PCB 9), 4-hydroxy-2',3,5-trichlorobiphenyl (4-OH PCB 34), and 4'-hydroxy 2,3',4,5'-tetrachlorobiphenyl (4'-OH PCB 68). Our results showed that 4-OH PCB 34 and 4'-OH PCB 68 were substrates for SULT2A1, and 4-OH PCB 34 exhibited substrate inhibition similar to that seen with the physiological substrate dehydroepiandrosterone (DHEA). Although the sulfation of 4-OH PCB 34 and 4'-OH PCB 68 represents a potential metabolic route for these compounds, these OHPCBs may also compete with other xenobiotic substrates as well as endogenous substrates for SULT2A1. The third OHPCB studied, 4'-OH PCB 9, was not a substrate for SULT2A1 but was an inhibitor of the enzyme. Thus, the interactions of OHPCBs with human SULT2A1 represent both a potential route of metabolism and a possible source of interference with sulfation reactions catalyzed by this enzyme. PMID- 17112229 TI - Site-specific arylation of rat glutathione s-transferase A1 and A2 by bromobenzene metabolites in vivo. AB - The hepatotoxicity of bromobenzene (BB) derives from its reactive metabolites (epoxides and quinones), which arylate cellular proteins. Application of proteomic methods to liver proteins from rats treated with a hepatotoxic dose of [14C]-BB has identified more than 40 target proteins, but no adducted peptides have yet been observed. Because such proteins are known to contain bromophenyl- and bromodihydroxyphenyl derivatives of cysteine, histidine, and lysine, the failure to observe modified peptides has been attributed to the low level of total covalent binding and to the "dilution" effect of multiple metabolites reacting at multiple sites on multiple proteins. In this work glutathione S transferase (GST), a well-known and abundant BB-target protein, was isolated from liver cytosol of rats treated with 14C-BB by use of a glutathione (GSH)-agarose affinity column and further resolved by reverse-phase high-performance liquid chromatography (HPLC) into subunits M1, M2, A1, A2 and A3. The subunits were identified by a combination of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), whole-molecule mass spectrometry, and peptide mass mapping and found to contain radioactivity corresponding to 0.01-0.05 adduct per molecule of protein. Examination of tryptic digests of these subunits by matrix assisted laser desorption ionization time-of-flight (MALDI-TOF) and electrospray ionization mass spectrometry (ESI-MS) failed to reveal any apparent adducted peptides despite observed sequence coverages up to 87%. However, use of HPLC linear ion-trap quadrupole Fourier transform mass spectrometry (LTQ-FTMS) to search for predicted modified tryptic peptides revealed peaks corresponding, with a high degree of mass accuracy, to a bromobenzoquinone adduct of peptide 89-119 in both GSTA1 and A2. The identity of these adducts and their location at Cys-111 was confirmed by tandem mass spectrometry (MS-MS). No evidence for the presence of any putative BB-adducts in GST M1, M2, or A3 was obtained. This work highlights the challenges involved in the unambiguous identification of reactive metabolite adducts formed in vivo. PMID- 17112230 TI - Photomutagenicity of anhydroretinol and 5,6-epoxyretinyl palmitate in mouse lymphoma cells. AB - Retinyl palmitate (RP) is frequently used as an ingredient in cosmetics and other retail products. We previously reported that, under UVA light irradiation, RP is facilely decomposed into multiple products, including anhydroretinol (AR) and 5,6 epoxyretinyl palmitate (5,6-epoxy-RP). We also determined that combined treatment of mouse lymphoma cells with RP and UVA irradiation produced a photomutagenic effect. In this study, we evaluated the photomutagenicity of AR and 5,6-epoxy-RP, in L5178Y/Tk+/- mouse lymphoma cells. Treatment of cells with AR or 5,6-epoxy-RP alone at 10 and 25 microg/mL for 4 h did not show a positive mutagenic response. However, because these doses did not induce the required amount of cytotoxicity for mouse lymphoma assay, we are unable to determine whether or not these two compounds are mutagenic. Treatment of cells with 1-25 microg/mL AR or 5,6-epoxy RP under UVA light (315-400 nm) for 30 min (1.38 mW/cm2) produced a synergistic photomutagenic effect. At 10 microg/mL (37.3 microM) AR with UVA exposure, the mutant frequency (MF) was about 3-fold higher than that for UVA exposure alone, whereas the MF for 25microg/mL (46.3microM) of 5,6-epoxy-RP + UVA was approximately 2-fold higher than that for UVA exposure alone. Compared with previous results for RP + UVA treatment, the potency of the induced phototoxicity and photomutagenicity was AR > RP > 5,6-epoxy-RP. To elucidate the underlying photomutagenic mechanism, we examined the loss of heterozygosity (LOH) at four microsatellite loci spanning the entire chromosome 11 for mutants induced by AR or 5,6-epoxy-RP. Most mutants lost the Tk+ allele, and more than 70% of the chromosome damage extended to 38 cM in chromosome length. AR + UVA induced about twice as many mutants that lost all four microsatellite markers from the chromosome 11 carrying the Tk+ allele as RP + UVA or 5,6-epoxy-RP + UVA. These results suggest that two of RP's photodecomposition products are photomutagenic in mouse lymphoma cells, causing events that affect a large segment of the chromosome. PMID- 17112231 TI - Comparison of p53 mutations induced by PAH o-quinones with those caused by anti benzo[a]pyrene diol epoxide in vitro: role of reactive oxygen and biological selection. AB - Polycyclic aromatic hydrocarbons (PAH) are one of the major carcinogens in tobacco smoke. They are metabolically activated through different routes to form either diol-epoxides, PAH o-quinones, or radical cations, each of which has been proposed to be an ultimate carcinogen. To study how PAH metabolites mutate p53, we used a yeast reporter gene assay based on p53 transcriptional activity. Colonies expressing wt p53 turn white (ADE +) and those expressing mutant p53 turn red (ADE -). We examined the mutagenicity of three o-quinones, benzo[a]pyrene-7,8-dione, benz[a]anthracene-3,4-dione, and dimethylbenz[a]anthracene-3,4-dione, and compared them with (+/-)-anti benzo[a]pyrene diol epoxide ((+/-)-anti-BPDE) within the same system. The PAH o quinones tested gave a dose-dependent increase in mutation frequency in the range of 0.160-0.375 microM quinone, provided redox-cycling conditions were used. The dominant mutations were G to T transversions (>42%), and the incidence of hotspot mutations in the DNA-binding domain was more than twice than that expected by a random distribution. The dependence of G to T transversions on redox cycling implicates 8-oxo-dGuo as the lesion responsible, which is produced under identical conditions (Chem. Res. Toxicol. (2005) 18, 1027). A dose-dependent mutation frequency was also observed with (+/-)-anti-BPDE but at micromolar concentrations (0-20 microM). The mutation pattern observed was G to C (63%) > G to A (18%) > G to T (15%) in umethylated p53 and was G to A (39%) > G to C (34%) > G to T (16%) in methylated p53. The preponderance of G mutations is consistent with the formation of anti-BPDE-N2-dGuo as the major adduct. The frequency of hotspots mutated by (+/-)-anti-BPDE was essentially random in umethylated and methylated p53, suggesting that 5'-CpG-3' islands did not direct mutations in the assay. These data suggest that smoking may cause mutations in p53 by formation of PAH o-quinones, which produce reactive oxygen species. The resultant 8-oxo-dGuo yields a pattern of mutations but not a spectrum consistent with that seen in lung cancer; we suggest that the emergence of the spectrum requires biological selection. PMID- 17112232 TI - Chlorine dioxide oxidation of guanosine 5'-monophosphate. AB - The reactions between aqueous ClO2 and guanosine 5'-monophosphate (5'-GMP) are investigated from pH 5.96 to 8.30. The decay of ClO2 follows mixed first-order and second-order kinetics. The addition of chlorite (0.01-0.05 M) to the reaction mixture suppresses the reaction rate and changes the observed decay of ClO2 to second-order. The reaction rates increase greatly with pH to give oxidized products. The second-order rate constant for the guanosine anion is 4.7 x 10(5 )M 1 s-1 and comprises a mixture of rate constants, k1k2/k-1. The ratio k1/k-1, with a calculated value of 2.4 x 10(-4), corresponds to the reversible reaction between ClO2 and the guanosine anion to generate ClO2- and the guanosyl radical. To determine k1/k-1 and k2, E values for guanosine and ClO2 are used as well as acid dissociation constants for guanosine and its radical. The value of k1 (1.1 x 10(5) M-1 s-1) represents the reaction between ClO2 and the guanosine anion as determined by initial rates. The second-order rate constant k2, with a value of 1.8 x 10(9 )M-1 s-1, represents the reaction between the guanosyl radical with a second molecule of ClO2 to generate a guanosyl-OClO adduct. The consumption of two mol of ClO2 per mol of 5'-GMP corresponds to a four-electron oxidation that gives ClO(2- )in the first step and HOCl in the second step. The 2',3',5'-tri-O acetylated derivative of guanosine is used to more easily separate guanosine from its ClO2 oxidation products. Imidazolone and monochlorinated imidazolone are identified as products of the reaction between ClO2 and guanosine. PMID- 17112233 TI - Cell growth inhibition and actin cytoskeleton disorganization induced by azaspiracid-1 structure-activity studies. AB - Azaspiracid-1 (AZA-1) is a marine toxin discovered 10 years ago. Since then, toxicologic studies have demonstrated that AZA-1 targets several organs in vivo, including the intestine, lymphoid tissues, lungs, and nervous system; however, the mechanism of action of AZA-1 remains unknown. Studies in vitro suggest that AZA-1 affects the actin cytoskeleton in nonadherent cells. We characterized the effects of AZA-1 on the cytoskeleton of adherent cells and on cell growth, an adhesion-dependent process in many cell types, and analyzed the structure dependency of this toxicity. Confocal and TIRF imaging of fluorescently labeled cytosketon showed that AZA-1 induced the rearrangement of stress fibers (actin filament bundles) and the loss of focal adhesion points in neuroblastoma and Caco 2 cells, without affecting the amount of polymerized actin. AZA-1 did not seem to alter the microtubule cytoskeleton, but it changed the cell shape and internal morphology observed by phase contrast imaging. Cell growth of lung carcinoma and neuroblastoma cells was inhibited by the toxin, as measured by a sulforhodamine B assay and BrdU incorporation to newly synthesized DNA. Fifteen different fragments and/or stereoisomers of AZA-1 were tested for cytoskeletal rearrangement and cell growth inhibition. Results showed that no fragment or stereoisomer had any activity, except for ABCD-epi-AZA-1, which conserved toxicity. AZA-1-induced reorganization of the actin cytoskeleton concurred with detachment and growth inhibition, three events that are probably related. PMID- 17112234 TI - Site-specific synthesis of oligonucleotides containing malondialdehyde adducts of deoxyguanosine and deoxyadenosine via a postsynthetic modification strategy. AB - Malondialdehyde (MDA) and its reactive equivalent, base propenal, are products of oxidative damage to lipids and DNA, respectively; they are mutagenic in bacterial and mammalian systems, and MDA is carcinogenic in rats. MDA adducts of deoxyguanosine (M1dG), deoxyadenosine (OPdA), and deoxycytidine (OPdC) have been characterized. We have developed site-specific syntheses of M1dG and OPdA adducted oligonucleotides that rely on a postsynthetic modification strategy. This work provides an alternative route to the M1dG adducted oligonucleotide and, to date, the only viable strategy for the site-specific synthesis of OPdA modified oligonucleotides. The stability of the modified oligonucleotides was examined by UV thermal melting studies (Tm). In contrast to the M1dG adduct, OPdA caused very little change in the Tm. PMID- 17112235 TI - A role for peroxymonocarbonate in the stimulation of biothiol peroxidation by the bicarbonate/carbon dioxide pair. AB - Peroxymonocarbonate (HCO4-) is an oxidant whose existence in equilibrium with hydrogen peroxide and bicarbonate has been known since the 1980s. More recently, peroxymonocarbonate has been proposed to mediate oxidative processes stimulated by the bicarbonate/carbon dioxide pair. To better understand this emerging biological oxidant, we re-examined the kinetics of its formation from hydrogen peroxide and bicarbonate/carbon dioxide by 13C NMR. Also, we studied its role in the accelerating effects of bicarbonate on biothiol (GSH and BSA-cysSH) peroxidation by kinetics and product analysis. The rate constants for peroxymonocarbonate formation and decay were estimated and Keq values determined (pH 7.2, at 25 and 37 degrees C; in the absence and presence of BSA and liposomes of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine phosphatidylcholine). Noteworthy is the fact the rate constant for peroxymonocarbonate formation estimated here (k1 approximately 10-2 M-1 s-1) was more than 1 order of magnitude higher than a previously reported value. Also, peroxymonocarbonate equilibrium was shown to be affected by BSA, liposomes, and a carbonic anhydrase mimetic. The Keq values determined in the absence and presence of BSA (0.35 and 0.48 M-1, respectively, at 37 degrees C) were employed to analyze the kinetics of BSA-cysSH and GSH peroxidation in the presence of bicarbonate (2-25 mM). A good fit of experimental data with simulations indicated that peroxymonocarbonate is the main species responsible for biothiol peroxidation in the presence of bicarbonate. The results indicate that peroxymonocarbonate is a feasible biological oxidant, in addition to supporting emerging data that the main physiological buffer is redox active. PMID- 17112236 TI - Mutagenesis of the supF gene of pSP189 replicating in AD293 cells cocultivated with activated macrophages: roles of nitric oxide and reactive oxygen species. AB - Dysregulated production of nitric oxide (NO*) and reactive oxygen species by inflammatory cells contributes to mutagenesis and carcinogenesis. We have characterized mutagenesis in the target supF gene of pSP189 replicating in AD293 cells cocultivated with mouse macrophage-like RAW264.7 cells activated with interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS). Activated macrophages produced substantial amounts of NO*, superoxide anion (O2*-), and hydrogen peroxide (H2O2) over 12-72 h periods. A time-dependent decrease in total cell number and a 3.7-fold increase in supF mutation frequency (MF), compared with unstimulated controls, were observed at 72 h. The increase in MF was effectively suppressed by N-methyl-L-arginine monoacetate (NMA), an NO* synthase inhibitor, and also by superoxide dismutase (SOD) and catalase (CAT); cotreatment with NMA and SOD/CAT suppressed mutagenesis by 87% at 72 h. Mutations in supF were mainly multiple sequence changes (47%) and single base pair substitutions (51%) following IFN-gammaLPS activation. Following cotreatment with NMA alone or together with SOD/CAT, however, single base pair substitutions were prevalent (70 and 85%); decreased multiple mutations were observed (24 and 11%). Almost all single base pair substitutions induced under all exposure conditions occurred at G:C base pairs (87.8-94.6%). Whereas those induced by all treatments consisted predominantly of G:C to T:A transversions, G:C to T:A and A:T to T:A transversions were less frequent following treatment with NMA alone or with SOD/CAT compared to those induced by activated macrophages without additional treatment. Our results strongly suggest that ONOO- or its derivatives generated by reaction of NO* with O2*- may have been a major contributor to the observed mutagenesis by the activated macrophages, and mitigating their effects might serve a preventive function in ameliorating cancer risks associated with prolonged inflammation. PMID- 17112237 TI - Chronic exposure to particulate chromate induces spindle assembly checkpoint bypass in human lung cells. AB - One of the hallmarks of lung cancer is chromosome instability (CIN), particularly a tetraploid phenotype, which is normally prevented by the spindle assembly checkpoint. Hexavalent chromium Cr(VI) is an established human lung carcinogen, and Cr(VI) induces tumors at lung bifurcation sites where Cr(VI) particles impact and persist. However, the effects of Cr(VI) on the spindle assembly checkpoint are unknown and little is known about prolonged exposure to particulate Cr(VI). Accordingly, we investigated particulate Cr(VI)-induced bypass of the spindle assembly checkpoint after several days of exposure in WHTBF-6 cells. We found that lead chromate indeed induces spindle assembly checkpoint bypass in human lung cells, as 72, 96, and 120 h treatments with 0.5 or 1 microg/cm2 lead chromate induced significant increases in the percentage of cells with aberrant mitotic figures. For example, treatment with 1 microg/cm2 lead chromate for 96 h induced 11, 12.3, and 14% of cells with premature anaphase, centromere spreading and premature centromere division, respectively. In addition, we found a disruption of mitosis with more cells accumulating in anaphase; cells treated for 96 h increased from 18% in controls to 31% in cells treated with lead chromate. To confirm involvement of the spindle assembly checkpoint, Mad2 expression was used as a marker. Mad2 expression was decreased in cells exposed to chronic treatments of lead chromate, consistent with disruption of the checkpoint. We also found concentration- and time-dependent increases in tetraploid cells, which continued to grow and form colonies. When cells were treated with chronic lead alone there was no increase in aberrant mitotic cells or polyploidy; however, chronic exposure to a soluble Cr(VI) showed an increase in aberrant mitotic cells and polyploidy. These data suggest that lead chromate does induce CIN and may be one mechanism in the development of Cr(VI)-induced lung cancer. PMID- 17112238 TI - Pro-oxidant activity of flavonoids induces EpRE-mediated gene expression. AB - Flavonoids are important bioactive dietary compounds. They induce electrophile responsive element (EpRE)-mediated expression of enzymes, such as NAD(P)H-quinone oxidoreductase (NQO1) and glutathione S-transferases (GSTs), which are major defense enzymes against electrophilic toxicants and oxidative stress. The induction of EpRE-mediated gene transcription involves the release of the transcription factor Nrf2 from a complex with Keap1, either by a direct interaction of the inducer with Keap1 or by protein kinase C (PKC)-mediated phosphorylation of Nrf2. The inhibition of PKC in Hepa1c1c7 cells, stably transfected with human NQO1-EpRE-controlled luciferase revealed that PKC is not involved in flavonoid-induced EpRE-mediated gene transcription. However, the ability of flavonoids to activate an EpRE-mediated response correlates with their redox properties characterized by quantum mechanical calculations. Flavonoids with a higher intrinsic potential to generate oxidative stress and redox cycling are the most potent inducers of EpRE-mediated gene expression. Modulation of the intracellular glutathione (GSH) level showed that the EpRE-activation by flavonoids increased with decreasing GSH and vice versa, supporting an oxidative mechanism. In conclusion, the pro-oxidant activity of flavonoids can contribute to their health-promoting activity by inducing important detoxifying enzymes, pointing to a beneficial effect of a supposed toxic chemical reaction. PMID- 17112240 TI - Chalcone inhibition of anthracycline secondary alcohol metabolite formation in rabbit and human heart cytosol. AB - Antineoplastic therapy with anthracyclines like doxorubicin (DOX) and daunorubicin (DNR) is limited by the possible development of a dose-related cardiomyopathy. Secondary alcohol metabolites like doxorubicinol (DOXol) and daunorubicinol (DNRol), formed by cytoplasmic two-electron reductases, have been implicated as potential mediators of anthracycline-induced cardiomyopathy. In the present study, we characterized the effects of 12 chalcones on the formation of anthracycline secondary alcohol metabolites by rabbit or human heart cytosol and compared them with those of quercetin and other flavonoids. Both chalcones and flavonoids inhibited DOXol or DNRol formation in isolated rabbit heart cytosol. Structure--activity relationships showed that inhibition by chalcones was determined primarily by the position of hydroxyl groups in their phenolic A and B rings. In particular, the presence of a hydroxyl group at C-4' in the A ring was an important determinant of the inhibitory activity of chalcones. Among chalcones, 2',4',2-trihydroxychalcone exhibited the highest inhibition of both DOXol and DRNol formation, but it proved less efficient than quercetin. Different results were obtained with isolated human heart cytosol: in the latter, 2',4',2 trihydroxychalcone and other hydroxychalcones inhibited both DOXol and DNRol formation, whereas quercetin and other flavonoids inhibited DNRol formation but failed to inhibit or slightly stimulated DOXol formation. These results identify chalcones as versatile inhibitors of the cytoplasmic reductases that convert anthracyclines to cardiotoxic secondary alcohol metabolites. PMID- 17112241 TI - Identification of glutathione-related quercetin metabolites in humans. AB - The glutathionylation of quercetin was investigated in murine hepatic suspensions, in the absence of chemically or enzymatically induced oxidative stress, and in human urine after the consumption of 200 g of cooked onions ( approximately 74 mg of quercetin). In murine hepatic suspensions, 22 metabolites, including glucuronide, sulfate, and glutathione conjugates of quercetin, were identified by LC/ESI-MS/MS. In total, eight glutathione conjugates were identified in these suspension, including three isomeric forms of monoglutathionyl quercetin, two isomers of monoglutathionyl quercetin glucuronide, and three isomers of glutathionyl methyl quercetin. Quinone forms of glutathionyl quercetin and glutathionyl methyl quercetin were also apparent in mass spectra. In humans, several glutathione-related metabolites of quercetin were identified in urine as mercapturic acids of common hydroxyphenylacetic acids generated by the microbial degredation of quercetin in the gut. These include mercaptic acids of dihydroxytoluene, dihydroxybenzaldehyde, dihydroxyphenylacetic acid, dihydroxycinnamic acid, and dihydroxyphenylpropionic acid. Our results suggest that glutathionylation of quercetin occurs in both murine hepatic suspensions and humans and indicate that under certain conditions, quercetin intermediates require inactivation through conjugation with glutathione. PMID- 17112239 TI - Translesion synthesis past the C8- and N2-deoxyguanosine adducts of the dietary mutagen 2-Amino-3-methylimidazo[4,5-f]quinoline in the NarI recognition sequence by prokaryotic DNA polymerases. AB - 2-Amino-3-methylimidazo[4,5-f]quinoline (IQ) is found in cooked meats and forms DNA adducts at the C8- and N2-positions of dGuo after appropriate activation. IQ is a potent inducer of frameshift mutations in bacteria and is carcinogenic in laboratory animals. We have incorporated both IQ-adducts into the G1- and G3 positions of the NarI recognition sequence (5'-G1G2CG3CC-3'), which is a hotspot for arylamine modification. The in vitro replication of the oligonucleotides was examined with Escherichia coli pol I Klenow fragment exo-, E. coli pol II exo-, and Sulfolobus solfataricus P2 DNA polymerase IV (Dpo4), and the extension products were sequenced by tandem mass spectrometry. Replication of the C8-adduct at the G3-position resulted in two-base deletions with all three polymerases, whereas error-free bypass and extension was observed at the G1-position. The N2 adduct was bypassed and extended by all three polymerases when positioned at the G1-position, and the error-free product was observed. The N2-adduct at the G3 position was more blocking and was bypassed and extended only by Dpo4 to produce an error-free product. These results indicate that the replication of the IQ adducts of dGuo is strongly influenced by the local sequence and the regioisomer of the adduct. These results also suggest a possible role for pol II and IV in the error-prone bypass of the C8-IQ-adduct leading to frameshift mutations in reiterated sequences, whereas noniterated sequences result in error-free bypass. PMID- 17112242 TI - Top-priority fragment QSAR approach in predicting pesticide aquatic toxicity. AB - In the framework of pesticide risk assessment, a fragment-based QSAR approach is presented to correlate LC50-96 h acute toxicity to the rainbow trout (Oncorhynchus mykiss). While there are other fragment-based modeling routes, our approach exploits the possibility of prioritizing fragments' contributions to toxicity. On the assumption that one fragment might be mainly responsible for the molecular toxicity, we developed a three-stage modeling strategy to select the most important moieties and to establish their priorities at a molecular level. This strategy was tested on a heterogeneous dataset containing 282 pesticides, collected under the EU-funded project Demetra. Quantitative toxicity prediction yielded good results for the training set (R2TR = 0.85) and the test set (R2TS = 0.75). The advantages and limitations of the current priority strategy are examined. PMID- 17112243 TI - QSAR prediction of estrogen activity for a large set of diverse chemicals under the guidance of OECD principles. AB - A large number of environmental chemicals, known as endocrine-disrupting chemicals, are suspected of disrupting endocrine functions by mimicking or antagonizing natural hormones, and such chemicals may pose a serious threat to the health of humans and wildlife. They are thought to act through a variety of mechanisms, mainly estrogen-receptor-mediated mechanisms of toxicity. However, it is practically impossible to perform thorough toxicological tests on all potential xenoestrogens, and thus, the quantitative structure--activity relationship (QSAR) provides a promising method for the estimation of a compound's estrogenic activity. Here, QSAR models of the estrogen receptor binding affinity of a large data set of heterogeneous chemicals have been built using theoretical molecular descriptors, giving full consideration to the new OECD principles in regulation for QSAR acceptability, during model construction and assessment. An unambiguous multiple linear regression (MLR) algorithm was used to build the models, and model predictive ability was validated by both internal and external validation. The applicability domain was checked by the leverage approach to verify prediction reliability. The results obtained using several validation paths indicate that the proposed QSAR model is robust and satisfactory, and can provide a feasible and practical tool for the rapid screening of the estrogen activity of organic compounds. PMID- 17112244 TI - Unique and common metabolites of thiamethoxam, clothianidin, and dinotefuran in mice. AB - The established neonicotinoid insecticides have chloropyridylmethyl (imidacloprid, thiacloprid, acetamiprid, and nitenpyram), chlorothiazolylmethyl (thiamethoxam or TMX and clothianidin or CLO) or tetrahydrofuranylmethyl (dinotefuran or DIN) substituents. We recently reported the metabolic fate of the chloropyridylmethyl neonicotinoids in mice as the first half of a comparative study that now considers the chlorothiazolylmethyl and tetrahydrofuranylmethyl analogues. TMX, CLO, two desmethyl derivatives (TMX-dm and CLO-dm), and DIN were administered ip to mice at 20 mg/kg for characterization of metabolites and pharmacokinetic analysis of brain, liver, plasma, and urine by HPLC/DAD and LC/MSD. Each compound is excreted 19-55% unmetabolized in urine within 24 h, and tissue residues are largely dissipated by 4 h. Thirty-seven metabolites of TMX, TMX-dm, CLO, and CLO-dm are identified by comparison with synthetic standards or their structures are proposed by molecular weights and 35Cl:37Cl ratios often supplemented by previous reports or sequence studies in which intermediates are readministered. A facile reaction sequence involves TMX --> TMX-dm or CLO --> CLO dm. CLO-dm, reported to be a contributor to TMX hepatocarcinogenesis in mice, is unexpectedly remethylated in part to CLO in brain. The nitrosoguanidine, aminoguanidine, and urea derivatives of the parent compounds are detected in the tissues and methylnitroguanidine, methylguanidine, and nitroguanidine in the urine. Chlorothiazolecarboxaldehyde from oxidative cleavage of TMX and CLO is quite persistent in brain, liver, and particularly plasma compared with chloropyridinecarboxaldehyde and tetrahydrofurancarboxaldehyde from the other neonicotinoids. Chlorothiazolecarboxylic acid is conjugated with glycine or glucuronic acid or converted to S-methyl and mercapturate derivatives. DIN metabolism involves nitro reduction, N-demethylation, N-methylene hydroxylation, and amine cleavage, and tetrahydrofuranylmethyl hydroxylation at the 2-, 4-, and 5-positions giving 29 tentatively identified metabolites. The diversity of biodegradable sites and multiple pathways insures against parent compound accumulation but provides intermediates reported to be active as nicotinic agonists and inducible nitric oxide synthase inhibitors. PMID- 17112246 TI - Icosaniobate: a new member of the isoniobate family. AB - Icosaniobate [Nb20O54]8- was synthesized by reacting [H4Nb6O19]4- with NO in tetrahydrofuran or MeNO2. A single-crystal X-ray diffraction study of its n tetrabutylammonium salt [a = 17.7284(18) A, b = 33.542(3) A, c = 34.316(2) A, Z = 4, and space group P22(1)2(1)] revealed a dimeric structure where two decaniobate ions are condensed sharing two terminal O atoms. Unlike that in [(NbW5O18)2O]4-, the Nb-O-Nb bridges in icosaniobate are bent. The nonlinear bridging reduces the maximum possible symmetry of the dimeric anion to mm2, which it closely approximates. PMID- 17112247 TI - True blue: blue-emitting aluminum(III) quinolinolate complexes. AB - Blue-emitting heteroleptic aluminum(III) bis(2-methyl-8-quinolinolate)phenolate complexes were synthesized. A tunable, blue-to-green emission is achieved by attaching electron-withdrawing modulators to the emisssive quinaldinate ligand. The electronic nature of modulator substituents attached to the position of the highest HOMO (highest occupied molecular orbital) density is used to modulate ligand HOMO levels to achieve effective emission tuning to obtain blue-emitting materials. Optical and electrochemical properties of the resulting complexes were investigated and compared to the results of density functional theory (DFT/B3LYP/6-31G*) studies. The resulting materials may find application as organic light-emitting device materials. PMID- 17112248 TI - Cyanide-bridged single-molecule magnet constructed by an octacoordinated [W(CN)6(bpy)]- anion. AB - Two cyanide-bridged WV-M [M = Mn(II) (1), Co(II) (2)] bimetallic clusters were prepared by self-assembling a new molecular precursor [W(CN)6(bpy)]- and the corresponding metal complexes. Compound 1 shows a tetranuclear W2Mn2 entity, consisting of a Jahn-Teller ion, Mn(III), which serves as an anisotropic source, while compound 2 exhibits a trimeric W2Co structure. Among them, compound 1 displays slow relaxation of the magnetization, which is typical of a single molecule magnet behavior. PMID- 17112249 TI - Size-controllable fabrication of noble metal nanonets using a TiO2 template. AB - We present herein a simple template method for preparing noble metal nanonets with defined sizes. The template utilized is a TiO2 nanotube (NT) array prepared by anodic oxidation of a pure titanium sheet in an electrolyte solution containing sodium fluoride. Uniform NTs with defined sizes are obtained by controlling the anodic potential. Gold nanonets are prepared by electrodepositing gold onto the template and then dissolving the TiO2 template in a 0.2 M HF solution. The pore size of the gold nanonet is determined by the TiO2 NT hole size. The formation mechanism of the nanonet is elucidated from field-emission scanning electron microscopy and transmission electron microscopy. Although a lot of reports have been presented on the synthesis of nanostructure materials, no work has been reported on the template synthesis of gold nanonets. This paper gives a simple and universal way to prepare noble metal nanonets. PMID- 17112250 TI - Helicity induction through hydrogen bonding and spontaneous resolution of a bimetallic nickel complex coordinated to an octahedral metalloligand. AB - A new bimetallic complex of nickel(II) coordinated to a molybdenum-containing metalloligand has been synthesized that forms a helical chain through hydrogen bonding and shows spontaneous resolution upon crystallization. PMID- 17112251 TI - A dinuclear Ni(II) complex with two types of intramolecular magnetic couplings: Ni(II)-Ni(II) and Ni(II)-TTF*+. AB - A dinuclear Ni(II) complex involving tetrathiafulvalene (TTF) radicals as ligands has been prepared and characterized, [Ni2(mu-Cl)2(L*+)2(I3)4(I2)3.(H2O)2.(C4H8O)3 (1), L = 4,5-bis(2-pyridylmethylsulfanyl)-4',5'-ethylenedithiotetrathiafulvalene. There are two types of intramolecular magnetic exchange interactions, namely one ferromagnetic Ni(II)-Ni(II) and one antiferromagnetic Ni(II)-TTF*+. This study is new in the respect of revealing a magnetic exchange interaction between a TTF*+ radical and a paramagnetic transition metal ion. This is due to the fact of a direct binding of the transition metal ion to the skeleton of the TTF*+ radical. PMID- 17112252 TI - Antimony-antimony bond formation by reductive elimination from a hafnium bis(stibido) complex. AB - The bis(stibido) complex CpCp*Hf(SbMes2)2 (2) was prepared and structurally characterized. Complex 2 reacts with 2 equiv of xylylisocyanide to give the bis insertion product CpCp*Hf[C(SbMes2)=N(2,6-MeC6H3)]2 (4). The reaction of 2 with oxidants (I2 and O2) or donors (carbon monoxide and diphenylacetylene) or thermolysis promotes the reductive elimination of Sb2Mes4. PMID- 17112253 TI - Where are the protons in alpha-[H(x)W(12)O(40)](8-x)- (x = 2-4)? AB - The equilibria and speciation of the proton cryptate polyoxometalate alpha (H2)W(12)O(40)]6- (1) were examined by NMR following the phase-transfer cation metathesis of aqueous Na(6)1 with Q+Br-/CH(2)Cl(2), leading to the isolation of the (n-Bu)4N+ (Q+) salts Q(6)1 and alpha-Q5[(H3)W(12)O(40)](Q(5)2). Several groups report salts of the protonated anions H(x)1 (x = 1 and 2) with no consensus on proton numbers or locations. Reported herein, a combination of 1H and 183W NMR evidence, elemental analysis, acid titration measurements, and H/D isotopomer assignments establishes that in nonaqueous media the internal cryptand cavity of 1 reversibly accommodates only one more proton to form 2. Because an external proton must transfer across the close-packed tungsten oxide surface of 1, which should constitute a substantial activation barrier, it is significant that the transformation is instantaneous by 1H NMR (1 equiv of HBr in CH(3)CN), whereas the reverse process is slow (t1/2 approximately 17.4 h; 1 equiv of Q+OH ). PMID- 17112254 TI - Mechanism of direct molecular oxygen insertion in a palladium(II)-hydride bond. AB - The mechanism of the direct insertion of molecular oxygen into a palladium hydride bond has been elucidated using quantum mechanics (B3LYP/LACVP** with the PBF continuum solvent model). The key step is found to be the abstraction of the hydrogen atom resulting in the formation of a PdI/HO2 (triplet) radical pair, which then proceeds to form a singlet palladium hydroperoxo species. Potential palladium(0) pathways were explored and were found to be inaccessible. The results are in agreement with recent experimental results and are consistent with our previously predicted mechanism for an analogue system. PMID- 17112255 TI - Synthesis and thermal decomposition of Zn(tda)H2O [tda = S(CH(2)COO)2(2-)]. AB - A novel two-dimensional coordination polymer Zn(tda)H2O [tda = S(CH(2)COO)2(2-)] was synthesized under hydrothermal conditions. The compound crystallized in monoclinic space group P2(1) with a = 16.4154(17) A, b = 5.2133(6) A, c = 16.4210(17) A, beta = 114.165(2) degrees , V = 1282.1(2) A3, and Z = 8. The structure features two-dimensional, noncentrosymmetric networks with a pseudohexagonal network of Zn2+ coordinated by tda and water molecules. Zn(tda)H2O decomposed at T > 300 degrees C to form a ZnO sponge with a surface area approximately 40 m2/g, which makes it an attractive precursor for nanoporous ZnO. PMID- 17112256 TI - [Me4N](Ni(II)(BEAAM)): a synthetic model for nickel superoxide dismutase that contains Ni in a mixed amine/amide coordination environment. AB - Nickel superoxide dismutase (NiSOD) is a metalloenzyme that converts O2*- into H2O2 and O2 by cycling between Ni(II) and Ni(III) oxidation states. Reduced NiSOD contains Ni(II) in a square-planar N2S2 coordination environment formed by two cysteinate S atoms, an amide N, and an amine N to Ni(II). [Me4N](Ni(II)(BEAAM)) represents the first NiN2S2 complex containing Ni in a mixed amine/amide environment. [Me4](Ni(II)(BEAAM)) contains Ni-S bonds at 2.177(2) and 2.137(2) A and Ni-N bonds at 1.989(7) and 1.858(6) A, which compare well with the metalloenzyme. Orange solutions of [Me(4)N](Ni(II)(BEAAM)) in MeCN are diamagnetic and stable toward O2 for weeks. A quasireversible Ni(II/III) redox couple is observed for [Ni(II)(BEAAM)](NMe4) at 0.12(1) V vs Ag/AgCl. These data suggest that NiSOD utilizes the mixed amine/amide ligands to modulate the Ni(II/III) redox couple to best match the O2*- reduction/oxidation couples while maintaining O2 stability. PMID- 17112257 TI - Prediction of anion distributions using Pauling's second rule. AB - Pauling's second crystal rule is shown to be able to predict in a general and simple way the distribution of anions in mixed oxyanion systems such as oxynitrides and oxyhalides with diverse compositions and structure types. Results are presented in a plot correlating the charge of the anions obtained from the observed occupancies in crystallographic positions with the calculated bond strength sums for these sites. PMID- 17112258 TI - Redox-controlled molecular flipper based on a chiral Cu complex. AB - A molecular bipaddled flipper based on a tetradentate chiral Cu complex has been designed. The paddling motion of this unprecedented molecular-scale machine can be controlled by reversible oxidation of the metal center. Kinetic and computational (density functional theory) analyses provide a detailed picture of the flipper motion at the molecular scale, rationalize the switching role of the metal-ion oxidation state, and pose the basis for the fine-tuning of the dynamic motion of this new class of molecular-scale devices. PMID- 17112259 TI - Electron transfer and ligand addition to atomic mercury cations in the gas phase: kinetic and equilibrium studies at 295 k. AB - Results are reported for experimental measurements of the room-temperature chemical reactions between ground-state Hg*+ ions and 16 important environmental and biological gases: SF6, CO, CO2, N2O, D2O, CH4, CH3F, O2, CH3Cl, OCS, CS2, NH3, C6F6, NO2, NO*, and C6H6. The inductively coupled plasma/selected-ion flow tube tandem mass spectrometer used for these measurements has provided both rate and equilibrium constants. Efficient electron transfer (>19%) is observed with CS2, NH3, C6F6, NO2, NO*, and C6H6, molecular addition occurs with D2O, CH4, CH3F, CH3Cl, and OCS, and SF6, CO, CO2, N2O, and O2 showed no measurable reactivity with Hg*+. Theory is used to explore the stabilities and structures of both the observed and unobserved molecular adducts of Hg*+, and reasonable agreement is obtained with experimental observations, given the uncertainties of the theory and experiments. A correlation is reported between the Hg*+ and proton affinities of the ligands investigated. Solvation of Hg*+ with formic acid was observed to increase the rate of electron transfer from NO* by more than 20%. PMID- 17112260 TI - Synthesis, structure, and electrochemical properties of a family of 2 (arylazo)phenolate complexes of ruthenium with unusual C-C coupling and N=N cleavage. AB - Reaction of 2-(4'-R-phenylazo)-4-methylphenols (R = OCH3, CH3, H, Cl, and NO2) with [Ru(dmso)(4)Cl2]affords a family of five ruthenium(III) complexes, containing a 2-(arylazo)phenolate ligand forming a six-membered chelate ring and a tetradentate ligand formed from two 2-(arylazo)phenols via an unusual C-C coupling linking the two ortho carbons of the phenyl rings in the arylazo fragment. A similar reaction with 2-(2'-methylphenylazo)-4-methylphenol with [Ru(dmso)(4)Cl2] has afforded a similar complex, in which one 2-(2' methylphenylazo)-4-methylphenolate ligand is coordinated forming a six-membered chelate ring, and the other two ligands have undergone the C-C coupling reaction, and the coupled species is coordinated as a tetradentate ligand forming a five membered N,O-chelate ring, a nine-membered N,N-chelate ring, and another five membered chelate ring. Reaction of 2-(2',6'-dimethylphenylazo)-4-methylphenol with [Ru(dmso)(4)Cl2] has afforded a complex in which two 2-(2',6' dimethylphenylazo)-4-methylphenols are coordinated as bidentate N,O-donors forming five- and six-membered chelate rings, while the third one has undergone cleavage across the N=N bond, and the phenolate fragment, thus generated, remains coordinated to the metal center in the iminosemiquinonate form. Structures of four selected complexes have been determined by X-ray crystallography. The first six complexes are one-electron paramagnetic and show rhombic ESR spectra. The last complex is diamagnetic and shows characteristic 1H NMR signals. All the complexes show intense charge-transfer transitions in the visible region and a Ru(III)-Ru(IV) oxidation on the positive side of SCE and a Ru(III)-Ru(II) reduction on the negative side. PMID- 17112261 TI - Metal-biradical chains from a high-spin ligand and bis(hexafluoroacetylacetonato)copper(II). AB - The synthesis, X-ray crystal structure, and magnetic studies of a rare example of organic/inorganic spin hybrid clusters extended in infinite ladder-type chain [Cu(C5F6HO2)2]7(C35H35N5O4)2 ([Cu(hfac)2]7(pyacbisNN)2, 2) formed by the reaction of a high spin nitronylnitroxide biradical C35H35N5O4 (pyacbisNN, 1) and bis(hexafluroacetylacetonate)copper(II) = Cu(hfac)2 are described. Single-crystal X-ray structure analysis revealed the triclinic P1 space group of 2 with the following parameters: a = 10.6191(4) A, b = 19.6384(7) A, c = 21.941(9) A, alpha = 107.111(7) degrees, beta = 95.107(8) degrees, gamma = 94.208(0) degrees , Z = 2. Each repeating unit in 2 carries a centrosymmetric cyclic six spin and a linear five spin cluster with four different copper coordination environments having octahedral and square planar geometries. These clusters are interconnected to form infinite chains which are running along the crystallographic b axis. The magnetic measurements show nearly paramagnetic behavior with very small variations over a large temperature range. The magnetic properties are thus result of complex competitions of many weak ferro- and antiferromagnetic interactions, which appear as small deviations from quite linear mu(eff) vs T dependence at low temperature. At high temperature (300-14 K), antiferromagnetic behavior dominates a little, while at very low temperature (14-2 K), a small increase of mu(eff) was observed. The magnetic susceptibility data are described by the Curie-Weiss law [chi = C/(T - theta)] with the optimal parameters C = 4.32 +/- 0.01 emuK/mol and theta = - 0.6 +/- 0.3 K, where C is the Curie constant and theta is the Weiss temperature. PMID- 17112262 TI - Pressure effect and crystal structure reinvestigations on the spin crossover system: [Fe(bt)2(NCS)2] (bt = 2,2'-bithiazoline) polymorphs A and B. AB - The crystal structure of [Fe(bt)2(NCS)2] (A) was determined by X-ray diffraction at 293 and at 150 K in order to analyze the structural changes associated with the spin transition. The space group is P1 with Z = 2 at both temperatures. Lattice constants are as follows: a = 8.5240(4), b = 11.0730(6), c = 12.5300(8) at 293 K and a = 8.1490(4), b = 11.4390(5), c = 12.1270(6) at 150 K. The iron(II) atom lies at the center of a distorted [FeN6] defined by two bt ligands arranged in a cis conformation. The two remaining coordination positions are occupied by two isothiocyanate anions. The average bond lengths of 2.159(4) A (293 K) and 1.951(2) A (150 K) clearly indicate the change in spin configuration. The trigonal distortion parameter phi has a value of 9.6 degrees and 5.5 degrees at 293 and 150 K, respectively. For A, DeltaV = DeltaV(SCO) = 28 A(3) per formula unit and is accompanied by a hysteresis of 10 K. chi(M)T vs T curves at atmospheric pressure for A show an abrupt spin transition with Tc downward arrow = 176 K and Tc upward arrow = 187 K. The thermodynamic parameters associated with the spin transition are DeltaH = 8.4 +/- 0.4 kJ mol(-1) and DeltaS = 46.5 +/- 3 J K mol(-1). The thermal dependence of the magnetic susceptibility at different pressures, 0.1-0.91 GPa, points out an unusual behavior, which can only be understood in terms of a crystallographic phase transition or a change in the bulk modulus of the complex. Polymorph B crystallizes in the C2/c space group with an average Fe-N bond length of 2.168(2) A and phi = 14.7 degrees at 293 K. B remains in the HS configuration even at pressures of 1.06 GPa. PMID- 17112263 TI - 2D and 3D supramolecular structures via hydrogen bonds and pi-stacking interactions in arylsulfonates of nickel and cobalt. AB - Five novel arylsulfonates of Ni and Co have been hydrothermally obtained and their structures determined by X-ray single-crystal diffraction. [Ni(Phen)(H2O)](1,5-NDS).2H2O (1) is a hydrogen-bonded supramolecular layered compound formed by self-assembly of tetraaqua-o-phenanthrolinenickel(II) cations and 1,5-naphthalenedisulfonate anions, [Ni(Phen)2(H2O)(1,5-NDS)] (2) and [Ni(Phen)2(H2O)(2,6-NDS)].2H2O (4) exhibit 2D structures via O-H...O and pi-pi (both perfect face-to-face and parallel-displaced) stacking interactions, [Co(Phen)2(H2O)(1,5-NDS)].2H2O (5) presents a 3D structure via O-H...O hydrogen bond layers and pi-pi parallel-displaced stacking interactions, and [Co(Phen)2(1,5-NDS)] (3) is the first example of a covalently bonded polymeric cobalt sulfonate. Its 1D structure comprises chains formed through bitopic 1,5 NDS linkers. Rates and selectivities for oxidation of organic sulfides with H2O2 were measured with 2-5. PMID- 17112264 TI - Synthesis, cytotoxic effect, and structure-activity relationship of Pd(II) complexes with coumarin derivatives. AB - We report the influence of the substituent at the N atom of the ligands on the synthesis, biological activity, and stability of Pd(II) complexes of the general formula PdL(2). The compounds adopt a cis or trans configuration with respect to the substituent at the nitrogen atom. Sterically hindered substituents promote the formation of trans isomers, whereas when the nitrogen atom is unsubstituted, cis isomers are formed. The compounds were characterized by elemental analysis, infrared and 1H NMR spectroscopies, and electrospray mass spectrometry. The complexes were also studied using X-ray diffraction and computational DFT methods. Both complexes cis-3a and trans-3c exhibit square-planar geometries around the Pd(II) atom. The cytotoxic effects of these complexes were examined on two human leukemia cell lines, HL-60 and NALM-6. Pd complex cis-3a showed significant cytotoxic activity. The effects exhibited by this complex were comparable to those reported for carboplatin. Loigand 2a was not cytotoxic. Computational analysis carried out at the PB/B3LYP/LACVP**//mPW1PW91/LanL2DZ level showed excellent correlation between the energy difference of the cis and trans isomers and the cytotoxic activity, rendering computations a useful predictive tool for the design of new drugs. PMID- 17112265 TI - CsR(R(6)CoI(12))2 (R = Gd, Er) and (CeI)0.26(Ce(6)MnI(9))2: two new structure types featuring R(6)Z clusters. AB - Compounds adopting two new structure types containing discrete lanthanide clusters have been found, CsR(R6CoI12)2 (R = Gd or Er) and (CeI)0.26(Ce6MnI9)2. CsEr(Er6CoI12)2 and CsGd(Gd6CoI12)2 were synthesized in reactions of CsI, RI3, CoI2, and R metals (3:19:6:23) heated to 750 degrees C for 500 h followed by slow cooling (0.1 degrees C/min). The X-ray crystal structure of CsEr(Er6CoI12)2 was solved in the Pa3 space group with a = 18.063(2) A at 250 K (Z = 4, R1 [I > 2sigma(I)] = 0.0459). (CeI)0.26(Ce6MnI9) was synthesized by combining KI, CeI3, MnI2, and Ce metal and heating to 850 degrees C for 500 h. The single-crystal X ray structure for (CeI)0.26(Ce6MnI9)2 was solved in the trigonal, P3 (147) space group with lattice parameters of a = 11.695(1) A and c = 10.8591(2) A (Z = 2, R1 [I > 2sigma(I)] = 0.0895). Elemental analyses (X-ray photoelectron spectroscopy (XPS) and atomic absorption spectroscopy (AAS)) were performed and show the absence of potassium in the structure. A disorder model was refined for the atoms in the large cavity. The magnetic susceptibility data for CsGd(Gd6CoI12)2 is consistent with strong intracluster ferromagnetic coupling, but intercluster antiferromagnetic coupling suppresses the susceptibility below 70 K. PMID- 17112266 TI - Electronic modulation of hyperpolarizable (porphinato)zinc(II) chromophores featuring ethynylphenyl-, ethynylthiophenyl-, ethynylthiazolyl-, and ethynylbenzothiazolyl-based electron-donating and -accepting moieties. AB - A series of conjugated (porphinato)zinc(II)-based chromophores structurally related to [5-(4-dimethylaminophenylethynyl)-15-(5-nitrothienyl-2-ethynyl)-10,20 bis(3,5-bis(3,3-dimethyl-1-butyloxy)phenyl)]zinc(II) were synthesized using metal catalyzed cross-coupling reactions involving [5-bromo-15-triisopropylsilylethynyl 10,20-diarylporphinato]zinc(II), [5-bromo-15-(4-dimethylaminophenylethynyl)-10,20 diarylporphinato]zinc(II), [5-(4-dimethylaminophenylethynyl)-15-ethynyl-10,20 diarylporphinato]zinc(II), and [5-(4-nitrophenylethynyl)-15-ethynyl-10,20 diarylporphinato]zinc(II), along with appropriately functionalized aryl, thienyl (or thiophenyl), thiazolyl, benzothiazolyl, and carbazolyl precursors. The linear and nonlinear optical properties of these asymmetrically 5,15-substitued-(10,20 diarylporphinato)zinc(II) chromophores that bear either 2-(9H-carbazol-9-yl) thiophen-5-yl-ethynyl, 4-dimethylaminophenylethynyl, or 2-(N,N diphenylamino)thiophen-5-yl-ethynyl electron-releasing groups and an electron withdrawing group selected from 2-formyl-thiophen-5-yl-ethynyl, 2-(2,2 dicyanovinyl)-thiophen-5-yl-ethynyl, 4-nitrophenylethynyl, 6-nitrobenzothiazol-2 yl-ethynyl, or 5-nitrothiazol-2-yl-ethynyl are reported. The dynamic hyperpolarizabilities of these compounds were determined from hyper-Rayleigh light scattering measurements carried out at a fundamental incident irradiation wavelength (lambda(inc)) of 1300 nm; these measured beta1300 values ranged from 690 --> 1400 x 10(-30) esu. These data (i) show that these neutral dipolar molecules express substantial beta1300 values, (ii) highlight that reductions in the magnitude of the aromatic stabilization energy of (porphinato)metal-pendant arylethynyl groups have a significant impact upon the magnitude of the molecular hyperpolarizability, and (iii) provide insights into advantageous design modifications for closely related structures having potential utility in long wavelength electrooptic applications. PMID- 17112267 TI - A tris(pyrazolyl) eta6-arene ligand that selects Cu(I) over Cu(II). AB - 1,3,5-Tris{2'-[(pyrazol-1-yl)methyl]phenyl}benzene, 4, and its complexes with Cu(I) and Ag(I) have been prepared and characterized. Both CuI4 and AgI4 triflate crystallize in the rhombohedral space group R3, with the cations and anions each exhibiting crystallographically imposed 3-fold (C3) symmetry. In both complexes, 4 behaves as a tris(pyrazolyl) eta6-arene ligand whose arms act as three-pronged tweezers to form chiral, propeller-like cations with pyramidal MN(pyrazole)3 coordination geometries. Centers of symmetry in the space group ensure that the crystals are racemates, with equal numbers of P,P,P and M,M,M enantiomers. In broad outline, each cation is shaped like a three-legged stool, with the metal ion centered at the top and pointed downward from a triangular N(pyrazole) plane toward the center of gravity (Cg) of the central benzene ring (a metal-endo conformation), which constitutes the bottom shelf of the stool. The Cu(I)...Cg and Ag(I)...Cg distances, 3.195(2) and 3.165(2) A, respectively, support the existence of an eta6 bonding interaction with Ag(I) and, to a lesser extent, with Cu(I). NMR data for AgI4 suggest rapid interconversion of this cation in solution between P,P,P and M,M,M enantiomers. Our inability to prepare any Cu(II) complexes with 4 is consistent with cyclovoltammetric results, which suggest that the ligand is more easily oxidized than Cu(I). PMID- 17112268 TI - Low-spin bis(2-methylimidazole)(octaethylporphyrinato)iron(III) chloride (perp [Fe(OEP)(2-MeHIm)(2)]Cl): a consequence of hydrogen bonding? AB - The synthesis and characterization of low-spin bis(2 methylimidazole)(octaethylporphyrinato)iron(III) chloride (perp[Fe(OEP)(2 MeHIm)2]Cl) is reported. The structure shows that the cation is a low-spin species with two imidazole ligands having a relative perpendicular orientation. The porphyrin core is very ruffled, which leads to shortened equatorial bonds of 1.974(4) A and slightly elongated axial Fe-N bond lengths of 2.005(10) A that are about 0.02 A shorter and 0.03 A longer, respectively, in comparison to bis imidazole ligated iron(III) species with parallel oriented axial ligands. A one dimensional hydrogen-bond chain is formed between chloride anions and uncoordinated imidazole nitrogen atoms. Compared with paral-[Fe(OEP)(2 MeHIm)2]ClO4, hydrogen bonding may play an important role in the differences in the two structures. Mossbauer spectra show broadened quadrupole doublets with quadrupole splittings of 1.81 mm/s at RT and 1.94 mm/s at 20 K. The isomer shift ranges from 0.26 to 0.36 mm/s. These confirm that the title complex is a low-spin iron(III) species with the ground state (dxy)2(dxz,dyz)3. Crystal data: monoclinic, space group P2(1)/c, a = 14.066(3) A, b, 20.883(4) A, c = 19.245(4) A, beta = 109.67 degrees , and Z = 4. PMID- 17112269 TI - Photophysical properties of ruthenium(II) tris(2,2'-bipyridine) complexes bearing conjugated thiophene appendages. AB - A small series of ruthenium(II) tris(2,2'-bipyridine) complexes has been synthesized in which ethynylated thiophene residues are attached to one of the 2,2'-bipyridine ligands. The photophysical properties depend on the conjugation length of the thiophene-based ligand, and in each case, dual emission is observed. The two emitting states reside in thermal equilibrium at ambient temperature and can be resolved by emission spectral curve-fitting routines. This allows the properties of the two states to be evaluated in both fluid butyronitrile solution and a transparent KBr disk. It is concluded that both emitting states are of metal-to-ligand charge-transfer (MLCT) character, and despite the presence of conjugated thiophene residues, there is no indication for a low-lying pi,pi*-triplet state that promotes nonradiative decay of the excited state manifold. A key feature of these systems is that the conjugation length imposed by the thiophene-based ligand helps to control the rate constants for both radiative and nonradiative decay from the two MLCT triplet states. PMID- 17112270 TI - Iron complexes of C- and N-methylated 2-aza-21-carbaporphyrin: NMR studies. AB - Insertion of iron(II) into methylated derivatives of N-confused porphyrins 2-aza 2-methyl-5,10,15,20-tetraphenyl-21-carbaporphyrin (MeCTPPH)H, 2-aza-5,10,15,20 tetraphenyl-21-methyl-21-carbaporphyrin (CTPPMe)H2, and 2-aza-2-methyl-5,10,15,20 tetraphenyl-21-methyl-21-carbaporphyrin (MeCTPPMe)H yielded N- or C-methylated high-spin iron(II) complexes (MeCTPPH)Fe(II)Br, (HCTPPMe)Fe(II)Br, and (MeCTPPMe)Fe(II)Br. One electron oxidation of (Me-CTPPH)Fe(II)Br using Br2, accompanied by deprotonation of a C(21)-H(21) fragment and formation of an Fe C(21) bond, produces an intermediate-spin, five-coordinate iron(III) complex (MeCTPP)Fe(III)Br. Simultaneously, a high-spin complex [(MeCTPPH)Fe(III)Br]+ was formed which preserved the side-on interaction between the metal ion and the inverted pyrrole ring. &[(MeCTPPH)Fe(III)Br]+ was also obtained by titration of (MeCTPP)FeIIIBr with TFA due to the C(21) protonation. A titration of (HCTPPMe)Fe(II)Br and (MeCTPPMe)Fe(II)Br with Br2 yielded solely corresponding high-spin iron(III) species [(HCTPPMe)Fe(III)Br+ and [(MeCTPPMe)Fe(III)Br+. Dioxygen reacts cleanly with (MeCTPPH)Fe(II)Br carbaporphyrin to form solely (MeCTPP)Fe(III)Br. The 1H NMR spectra of paramagnetic iron(II) and iron(III) complexes were examined. The characteristic patterns of pyrrole, C-methyl, and N methyl resonances were found diagnostic of the ground electronic state of iron and the coordinating nature of the N-confused pyrrole. The characteristic C-Me resonances occur in a unique window (520-420 ppm) for iron(III) C-methylated N confused porphyrins which remains in contrast with relatively small values found for iron(II) C-methylated derivatives (50-80 ppm). PMID- 17112271 TI - Midrange affinity fluorescent Zn(II) sensors of the Zinpyr family: syntheses, characterization, and biological imaging applications. AB - The syntheses and photophysical characterization of ZP9, 2-{2-chloro-6-hydroxy-3 oxo-5-[(2-{[pyridin-2-ylmethyl-(1H-pyrrol-2 ylmethyl)amino]methyl}phenylamino)methyl]-3H-xanthen-9-yl}benzoic acid, and ZP10, 2-{2-chloro-6-hydroxy-5-[(2-{[(1-methyl-1H-pyrrol-2-ylmethyl)pyridin-2 ylmethylamino]methyl}phenylamino)methyl]-3-oxo-3H-xanthen-9-yl}benzoic acid, two asymmetrically derivatized fluorescein-based dyes, are described. These sensors each contain an aniline-based ligand moiety functionalized with a pyridyl-amine pyrrole group and have dissociation constants for Zn(II) in the sub-micromolar (ZP9) and low-micromolar (ZP10) range, which we define as "midrange". They give approximately 12- (ZP9) and approximately 7-fold (ZP10) fluorescence turn-on immediately following Zn(II) addition at neutral pH and exhibit improved selectivity for Zn(II) compared to the di-(2-picolyl)amine-based Zinpyr (ZP) sensors. Confocal microscopy studies indicate that such asymmetrical fluorescein based probes are cell permeable and Zn(II) responsive in vivo. PMID- 17112272 TI - SHG active crystals of a remote functionalized achiral NLO-phore assembled through zinc(II) complexation. AB - An achiral nonlinear optical chromophore with a "remote functionality" that can act as a ligand is developed on the basis of 4-nitroaniline derivatized with pyridine. The molecules are assembled through complexation with simple achiral zinc(II) salts and the H-bond network mediated by the counterions, to generate noncentrosymmetric materials exhibiting optical second harmonic generation (SHG). The crystal structures of the new complexes are determined; the counterion strongly influences the ligand orientations and lattice structure. SHG of the microcrystalline materials is investigated. Correlation between the structure and SHG is rationalized using semiempirical quantum chemical estimation of the hyperpolarizabilities of molecules and molecular clusters. The metal complexation plays a significant role in molecular assembly but affects the SHG very little, enabling simplified analysis of the bulk property in terms of molecular responses. Organization of remote functionalized molecules by metal ion complexation thus offers a convenient approach to the rational design of quadratic NLO materials. PMID- 17112273 TI - Molecular pairs and a propeller containing quadruply bonded dimolybdenum units linked by polyamidate ligands. AB - Two molecular pairs [Mo2(DAniF)3]2[N,N'-diethylterephthalamidate] (1) and [Mo2(DAniF)3]2{1,3-C6H4[C(O)NP]} (2) where DAniF = N,N'-di-p-anisylformamidinate) and the propeller ([Mo2(DAniF)3]3{1,3,5-C6H3[C(O)NPh]3} (3)) have been prepared in good yield and high purity by directly combining Mo2(DAniF)3(O2CCH3) with the corresponding polyamidates. Electrochemical measurements of these complexes show unresolved redox waves, which indicate that the dimetal centers are only electronically weakly coupled. Compound 1 was chemically oxidized by ferrocenium tetrafluoroborate to the two-electron oxidation product 4, where one electron was removed from each of the [Mo2] units. The hyperfine coupling in the EPR spectrum (A = 22 x 10-4 cm-1 ) suggests that 4 is an electron-trapped species with one electron residing on each of the two dimolybdenum units, as suggested also by electrochemical measurements. PMID- 17112274 TI - Removing the sting from the tail: reversible protonation of scorpionate ligands in cobalt(II) tris(carbene)borate complexes. AB - Low-temperature deprotonation of the phenylborane dications, PhB(RIm)3OTf2 (R = tBu, Mes), followed by in situ reaction with CoCl2(thf)1.5, results in the formation of the four-coordinate complexes, kappa3-PhB(RIm)3CoCl, in which the metal is supported by tripodal N-heterocyclic carbene-based ligands. The chloride complexes are exceptionally sensitive to acid and can be reversibly protonated to form the zwitterions kappa2-{PhB(RIm)2(RIm.H)}CoCl2. This unexpected reactivity is attributed to the highly basic nature of the tris(carbene)borate ligands. Reaction of the chloride complexes with methylating reagents results in products that depend on the N-heterocyclic carbene substituent. For R = tBu, the four coordinate high-spin complex, kappa3-PhB(tBuIm)3CoMe, is formed, while for R = Mes, reduction to a multitude of complexes occurs. PMID- 17112275 TI - Rational design of 0D, 1D, and 3D open frameworks based on tetranuclear lanthanide(III) sulfonate-phosphonate clusters. AB - Hydrothermal reactions of lanthanide(III) salts with m-sulfophenylphosphonic acid (H3L1) and 1,10-phenanthroline (phen) or N,N'-piperazinebis(methylenephosphonic acid) (H4L2) afforded six novel lanthanide(III) sulfonate-phosphonates based on tetranuclear clusters, namely, [La(2)(L1)2(phen)4(H2O)].4.5H2O (1), [Ln2(L1)2(phen)2(H2O)5].3H2O (Ln = Nd, 2; Eu, 3; Er, 4), and [Ln2(HL1)(H2L2)2(H2O)4].8H2O (Ln = La, 5; Nd, 6). Compounds 2-4 contain discrete tetranuclear lanthanide(III) cluster units in which four lanthanide(III) ions are bridged by two tridentate and two tetradentate phosphonate groups. In compound 1, the tetranuclear clusters are further interconnected into a 1D chain through the coordination of the sulfonate groups. The structures of compounds 5 and 6 can be viewed as a 3D architecture based on a different types of tetranuclear cluster units that are interconnected by bridging H2L2 anions. In the tetranuclear clusters of compounds 5 and 6, the four lanthanide(III) centers are interconnected by only two HL1 ligands. Compound 2 is a luminescent material in the near-IR region, whereas compound 3 displays a strong luminescent emission band in the red-light region. Magnetic property measurements of compounds 2-4 and 6 indicate that there are strong antiferromagetic interactions between magnetic centers within the cluster units. PMID- 17112276 TI - Electronic structure and excited states of rhenium(I) amido and phosphido carbonyl-bipyridine complexes studied by picosecond time-resolved IR spectroscopy and DFT calculations. AB - UV-vis absorption and picosecond time-resolved IR (TRIR) spectra of amido and phosphido complexes fac-[Re(ER2)(CO)3(bpy)] (ER2 = NHPh, NTol2, PPh2, bpy = 2,2' bipyridine, Tol = 4-methylphenyl) were investigated in conjunction with DFT and TD-DFT calculations in order to understand their ground-state electronic structure, low-lying electronic transitions and excited-state character and dynamics. The HOMO is localized at the amido/phosphido ligand. Amide and phosphide ligands are sigma-bonded to Re, the pi interaction being negligible. Absorption spectra show a weak band at low energies (1.7-2.1 eV) that arises from essentially pure ER(2) --> bpy ligand-to-ligand charge transfer (LLCT). The lowest excited state is the corresponding triplet, (3)LLCT. Low triplet energies and large distortions diminish the excited-state lifetimes to 85 and 270 ps for NHPh and NTol(2), respectively, and to ca. 30 ps for PPh2. nu(CO) vibrations undergo only very small ( bpy MLCT character, is a unique feature of the amido/phoshido complexes, whose lowest excited state can be viewed as containing a highly unusual aminyl/phosphinyl radical-cationic ligand. For comparison, the amino and phosphino complexes fac-[Re(NHPh(2))(CO)3(bpy)]+ and fac [Re(PPh3)(CO)3(bpy)]+ are shown to have the usual Re --> bpy (3)MLCT lowest excited states, characterized by upshifted nu(CO) bands. PMID- 17112277 TI - Metal binding to bipyridine-modified PNA. AB - Substitution of natural nucleobases in PNA oligomers with ligands is a strategy for directing metal ion incorporation to specific locations within a PNA duplex. In this study, we have synthesized PNA oligomers that contain up to three adjacent bipyridine ligands and examined the interaction with Ni2+ and Cu2+ of these oligomers and of duplexes formed from them. Variable-temperature UV spectroscopy showed that duplexes containing one terminal pair of bipyridine ligands are more stable upon metal binding than their nonmodified counterparts. While binding of one metal ion to duplexes that contain two adjacent bipyridine pairs makes the duplexes more stable, additional metal ions lower the duplex stability, with electrostatic repulsions being, most likely, an important contributor to the destabilization. UV titrations showed that the presence of several bipyridine ligands in close proximity of each other in PNA oligomers exerts a chelate effect. A supramolecular chelate effect occurs when several bipyridines are brought next to each other by hybridization of PNA duplexes. EPR spectroscopy studies indicate that even when two Cu2+ ions coordinate to a PNA duplex in which two bipyridine pairs are next to each other, the two metal-ligand complexes that form in the duplex are far enough from each other that the dipolar coupling is very weak. EXAFS and XANES show that the Ni2+-bipyridine bond lengths are typical for [Ni(bipy)2]2+ and [Ni(bipy)3]2+ complexes. PMID- 17112278 TI - Mixed-valent selenidoantimonates(III,V) with transition-metal complexes as counterions: solvothermal syntheses and characterization of [M(dien)2]2Sb4Se9 (M = Mn, Fe), [Co(dien)2]2Sb2Se6, and [Ni(dien)2]2Sb2Se5. AB - Novel selenidoantimonate compounds [M(dien)2]2Sb4Se9 [M = Mn (1), Fe (2)], [Co(dien)2]2Sb2Se6 (3), and [Ni(dien)2]2Sb2Se5 (4) (dien = diethylenetriamine) were solvothermally synthesized and characterized. The unique features of compounds 1-3 are the mixed-valent anionic structures constructed by the Sb(III)Se3 trigonal pyramid and Sb(V)Se4 tetrahedron. Three Sb(III)Se3 pyramids share common corners, forming a heterocyclic Sb3Se6 moiety, and the Sb3Se6 moieties are further connected with Sb(V)Se4 tetrahedra to form the novel one dimensional [Sb4Se9(4-)]n anionic chain in 1 and 2. The discrete [Sb2Se6]4- anion in 3 is formed by an Sb(III)Se3 trigonal pyramid and an Sb(V)Se4 tetrahedron sharing a common corner. The [Sb2Se5]4- anion in 4 is composed of two Sb(III)Se3 trigonal pyramids connected in the same manner as the [Sb2Se6]4- anion. The mixed valent [Sb4Se9(4-)]n and [Sb2Se6]4- anions were not observed before. The synthesis and solid-state structural studies of the title compounds show that the transition-metal complexes exhibit different structure-directing effects on the formation of selenidoantimonates in dien. Extensive N-H...Se hydrogen bonds are observed between cations and anions in compounds 1-4, resulting in three dimensional network structures. Optical and thermal properties of the compounds are reported. PMID- 17112279 TI - Pressure-induced internal redox reaction of Cs2[PdI4].I2, Cs2[PdBr4].I2, and Cs2[PdCl4].I2. AB - The pressure-induced redox reaction within the system Cs2[Pd2+I4].I2/Cs2[Pd4+I6] was investigated by means of powder X-ray diffraction. Analogous high-pressure X ray diffraction experiments were performed on the isostructural compounds Cs2[PdX4].I2 (X = Cl, Br). Additionally, the phase transition of Cs2[PdBr4].I2 to Cs2[PdBr4I2] was characterized by means of Raman scattering experiments as well as theoretical calculations based on density functional theory. On the basis of experimentally determined crystal structure data, a pathway for the topology of the redox reactions was developed and outlined. PMID- 17112280 TI - Extremely long axial Cu-N bonds in chiral one-dimensional zigzag cyanide-bridged Cu(II)(-)Ni(II) and Cu(II)(-)Pt(II) bimetallic assemblies. AB - Preparations, crystal structures, and spectral and magnetic properties of two new chiral one-dimensional cyano-bridged coordination polymers, [Cu(II)L2]M(II)(CN)].2H2O (M(II) = Ni(II) (1) and Pt(II) (2), L = trans cyclohexane-(1R,2R)-diamine) have been presented. Complex 1 crystallizes in the monoclinic P2(1) space group with a = 9.864(4) A, b = 15.393(8) A, c = 7.995(4) A, beta = 110.32(3) degrees , V = 1138.4(10) A3, and Z = 2, while 2 crystallizes in the monoclinic P2(1) space group with a = 9.899(3) A, b = 15.541(4) A, c = 8.102(2) A, beta = 111.02(2) degrees , V = 1163.6(5) A3, and Z = 2. The unique zigzag cyano-bridged chains along the crystallographic b axis consist of alternate chiral [CuL(2)]2+ cations and square-planar [M(CN)4]2- anions. One side of the axial Cu-N(triple bond C) bond distances are 2.324(6) and 2.34(1) A with Cu-N[triple bond]C angles of 137.8(6) degrees and 138.2(9) degrees for 1 and 2, respectively. On the other hand, the opposite side of the axial Cu-N(triple bond C) bond distances are 3.120(8) and 3.09(1) A with significantly large bent Cu N[triple bond]C angles of 97.9(5) degrees and 96.8(7) degrees for 1 and 2, respectively. The novel axial bonding features of extremely long semi coordination Cu-N bonds are attributed to coexistence of pseudo-Jahn-Teller elongation and electrostatic interaction in the unique zigzag cyano-bridged chains. The characteristic bonding features with overlap between small 3d (Ni(II)) or large 5d (Pt(II)) and 3d (Cu(II)) orbitals results in larger shifts in XPS peaks of not only Cu2p(1/2) and Cu2p(3/2) but also Ni2p(1/2) and Ni2p(3/2) for 1 than those of 2, which is also consistent with weak antiferromagnetic interactions with Weiss constants of -5.31 and -5.94 K for 1 and 2, respectively. The d-d, pi-pi*, and CT bands in the electronic, CD, and MCD spectra for 1 and 2 in the solid state at room temperature are discussed from the viewpoint of magneto-optical properties. PMID- 17112281 TI - Synthesis and characterization of water-operative cationic and anionic metal-ion activated molecular receptors for aromatic anions. AB - Two new, octadentate, water-soluble, macrocyclic ligands, 1,4,7,10-tetrakis((2S) (-)-2-hydroxy-3-[3'-(N,N,N-trimethylammonium)-phenoxy]-propyl)-1,4,7,10 tetraazacyclododecane tetratriflate, ((S)-tmappc12 triflate, L1 triflate) and 1,4,7,10-tetrakis((2S)-(-)-2-hydroxy-3-[2'-sulfo-4'-methylphenoxy]-propyl) 1,4,7,10-tetraazacyclododecane, ((S)-sthmppc12, L2H4) have been prepared with a view to using them to study anion sequestration in aqueous solution. Their pKa and metal-ion binding constant values with a range of alkaline earth, transition, and post-transition metals are reported. The eight-coordinate, water-soluble Cd(II) complexes of (L1)4+ and (L2)4-, [CdL1](CF3SO3)6 and (NH4)2-[CdL2], the former cationic and the latter anionic, have both been shown to be capable of acting as anion receptors in aqueous solution. The binding constant values (log(K/M-1) given in parentheses) for binding by the cationic receptor to a range of aromatic anions in water are p-nitrophenolate (1.7), p-formylphenolate (2.1), p-nitrobenzoate (3.0), p-aminobenzoate (4.5), p-dimethylaminobenzoate (>4.5), D- and L-tryptophanate (1.6, 2.2), phenoxyacetate (2.1), and acetate (2.3). With the anionic receptor, nonzero binding constants were only measurable for p nitrobenzoate (approximately 0.4), p-aminobenzoate (2.0), and p dimethylaminobenzoate (1.8). By reference to the X-ray determined structures of related, but water-insoluble inclusion complexes, anion retention is thought to occur within a hydrophobic cavity, with four convergent hydroxy groups at its base, which develops in (L1)4+ and (L2)4- through the juxtapositioning of aromatic rings that occurs as a consequence of octadentate coordination. PMID- 17112282 TI - Oxo- and oxoperoxo-molybdenum(VI) complexes with aryl hydroxamates: synthesis, structure, and catalytic uses in highly efficient, selective, and ecologically benign peroxidic epoxidation of olefins. AB - A solution obtained by dissolving MoO3 in H2O2 reacts separately with secondary hydroxamic acids (viz., N-benzoyl N-phenyl hydroxamic acid (BPHAH), N-benzoyl N ortho-, -meta-, -para-tolyl hydroxamic acids, (BOTHAH, BMTHAH, and BPTHAH, respectively), and N-cinnamoyl N-phenyl hydroxamic acid (CPHAH) affording [MoO(O2)(BPHA)2] (1), [MoO(O2)(BOTHA)2] (2), [MoO(O2)(BMTHA)2] (3), [MoO(O2)(BPTHA)2] (4), and [Mo(O)2(CPHA)2](5), respectively. The O and O2 are situated cis to each other in 2-4, but in each case, they are disordered and distributed over four sites. This disorder does not exist in the 6-coordinate cis dioxo complex 5, to which crude MoO(O2)(CPHA)2 (5') was converted during recrystallization. An aqueous molybdate solution readily reacts with all those hydroxamic acids producing [Mo(O)2(hydroxamate)2] (6). While 2, 3, and 4 possess a very distorted pentagonal bipyramidal structure, 5 has a distorted octahedral geometry. In the solid state, as well as in solution, 5 exists as two apparently enantiomerically related molecules differing in the orientation of the pendant phenyl rings. To emphasize that the formation and structural uniqueness of 5 compared to 1-4 is caused by the influence of the cinnamoyl residue, one compound of the 6 series, namely, [Mo(O)2(BPHA)2] (6A), was structurally characterized to prove directly that the special stereochemical properties of 5 rely on the special electronic structure of CPHA- ligand. Complexes 1-5, as well as 6, show high potential and selectivity as catalysts in the epoxidation of olefins at room temperature in the presence of NaHCO3 as a promoter and H2O2 as a terminal oxidant. A comparative epoxidation study has been performed to determine the relative efficiency of the catalysts. To make the epoxidation method cost effective, a study to optimize the use of H2O2 has also been performed. To obtain evidence in favor of our suggested mechanism to this homogeneous olefin --> epoxide conversion, it was necessary to synthesize a peroxo-rich compound, namely, [MoO(O2)2BMTHA]- (7), but the attempted synthesis culminated in the isolation of [MoO(O2)2(C6H5COO)]- (8), obviously, via the hydrolysis of coordinated BMTHA. PMID- 17112283 TI - Electron deficient carbon-titanium triple bonds: formation of triplet XC/TiX3 methylidyne complexes. AB - Laser-ablated titanium atoms react with CX4 (X = F and Cl) to produce triplet state XC/TiX3 complexes trapped in an argon matrix. Products are identified by their infrared spectra and comparison to theoretically predicted vibrations. Density functional theory calculations converge to C(3v) symmetry structures for these lowest-energy products. The two unpaired electrons in the carbon 2p orbitals are shared with empty titanium d orbitals leading to degenerate singly occupied pi molecular orbitals and an electron-deficient triple bond between the carbon and titanium centers, on the basis of DFT bonding analysis and spin density calculations. The carbon-titanium distances are near typical C=Ti double bond lengths, and the C-X bonds in the resulting products are shorter than in the CX4 precursors. It appears that X lone-pair conjugation contributes to the C-X bond strength and somewhat to the C-Ti bond, and Cl does better in this regard than F. PMID- 17112284 TI - A systematic resolution of sulfur in reticulated vitreous carbon using X-ray absorption spectroscopy. AB - Sulfur K-edge X-ray absorption spectroscopy (XAS) was used to characterize the approximately 0.1% sulfur found both in native reticulated vitreous carbon (RVC) foam and in RVC oxidatively modified using 0.2 M KMnO4 in 2 M H2SO4. Sulfur valences and functional groups were assessed using K-edge XAS spectral curve fitting and employing explicit sulfur compounds as models. For native RVC, these were episulfide (approximately 3%), thianthrene (approximately 9%), disulfide (approximately 10%), sulfenate ester (approximately 12%), benzothiophene (approximately 24%), N,N'-thiobisphthalimide (approximately 30%), alkyl sulfonate (approximately 1.2%), alkyl sulfate monoester (approximately 6%), and sulfate dianion (approximately 6%). Permanganate oxidation of RVC diminished sulfenic sulfur to approximately 9%, thianthrenic sulfur to approximately 7%, and sulfate dianion to approximately 1% but increased sulfate monoester to approximately 12%, and newly produced sulfone (approximately 2%) and sulfate diester (approximately 5%). A simple thermodynamic model was derived that allows proportionate functional group comparisons despite differing (approximately +/-15%) total sulfur contents between RVC batches. The limits of accuracy in the XAS curve fitting analysis are discussed in terms of microenvironments and extended structures in RVC carbon that cannot be exactly modeled by small molecules. Sulfate esters cover approximately 0.15% of the RVC surface, increasing to approximately 0.51% following permanganate/sulfuric acid treatment. The detection of episulfide directly corroborates a proposed mechanism for the migration of elemental sulfur through carbon. PMID- 17112285 TI - Three autocatalysts and self-inhibition in a single reaction: a detailed mechanism of the chlorite-tetrathionate reaction. AB - The chlorite-tetrathionate reaction has been studied spectrophotometrically in the pH range of 4.65-5.35 at T = 25.0 +/- 0.2 degrees C with an ionic strength of 0.5 M, adjusted with sodium acetate as a buffer component. The reaction is unique in that it demonstrates autocatalysis with respect to the hydrogen and chloride ion products and the key intermediate, HOCl. The thermodynamically most-favorable stoichiometry, 2S(4)O(6)2- + 7ClO2- + 6H2O --> 8SO(4)2- + 7Cl- + 12H+, is not found. Under our experimental conditions, chlorine dioxide, the chlorate ion, or both are detected in appreciable amounts among the products. Initial rate studies reveal that the formation of chlorine dioxide varies in an unusual way, with the chlorite ion acting as a self-inhibitor. The reaction is supercatalytic (i.e., second order with respect to autocatalyst H+). The autocatalytic behavior with respect to Cl- comes from chloride catalysis of the chlorite-hypochlorous acid and hypochlorous acid-tetrathionate subsystems. A detailed kinetic study and a model that explains this unusual kinetic behavior are presented. PMID- 17112286 TI - Reactivity and equilibrium thermodynamic studies of rhodium tetrakis(3,5 disulfonatomesityl)porphyrin species with H2, CO, and olefins in water. AB - Aqueous (D2O) solutions of tetrakis(3,5-disulfonatomesityl)porphyrin rhodium(III) aquo/hydroxo complexes ([(TMPS)Rh(III)(D2O)2]-7 (1), [(TMPS)Rh(III)(OD)(D2O)]-8 (2), and [(TMPS)Rh(III)(OD)2]-9 (3)) react with hydrogen (D2) to form an equilibrium distribution with a rhodium hydride ([(TMPS)Rh-D(D2O)]-8 (4)) and a rhodium(I) complex ([(TMPS)Rh(I)(D2O)]-9 (5)). Equilibrium constants (298 K) are measured that define the distribution for all five of these (TMPS)Rh species in this system as a function of the dihydrogen (D2) and hydrogen ion (D+) concentrations. The hydride complex [(TMPS)Rh-D(D2O)]-8 is a weak acid in D2O (Ka(298 K) = 4.3 x 10(-8)). Steric demands of the TMPS porphyrin ligand prohibit formation of a Rh(II)-Rh(II)-bonded complex, related rhodium(I)-rhodium(III) adducts, and intermolecular association of alkyl complexes which are prominent features of the rhodium tetra(p-sulfonatophenyl)porphyrin ((TSPP)Rh) system. The rhodium(II) complex ([(TMPS)Rh(II)(D2O)]-8) reacts with water to form hydride and hydroxide complexes and is not observed in D2O. The (TMPS)Rh-OD and (TMPS)Rh-D bond dissociation free energies (BDFE) are virtually equal and have a value of approximately 60 kcal mol(-1). Reactions of [(TMPS)Rh-D(D2O)]-8 in water with CO and olefins produce rhodium formyl and alkyl complexes which have equilibrium thermodynamic values comparable to the values for the corresponding substrate reactions of [(TSPP)Rh-D(D2O)]-4. PMID- 17112287 TI - Spacially confined M2 centers (M = Fe, Co, Ni, Zn) on a sterically bulky binucleating support: synthesis, structures and ethylene oligomerization studies. AB - Two new bulky aryl-bridged pyridyl-imine compartmental (pro)ligands, 2,6-{(2,6-i Pr(2)C6H3)N=C(Me)C5H3N}2C6H3Y (Y = H L1, OH L2-H), have been prepared in moderate to good overall yields via a Stille-type cross-coupling approach. The molecular structure of L2-H reveals a transoid configuration within the pyridyl-imine units with a hydrogen-bonding interaction maintaining the phenol coplanar with one of the adjacent pyridine rings. The interaction of 2 equiv of MX2 with L1 in n-BuOH at 110 degrees C gives the binuclear complexes, [(L1)M2X4] (M = Fe, X = Cl (1a); M = Co, X = Cl (1b); M = Ni, X = Br (1c); M = Zn, X = Cl (1d)), in which the metal centers adopt distorted tetrahedral geometries and occupy the two pyridyl imine cavities in L1. In contrast, deprotonation of L2-H occurs upon reaction with 2 equiv of MX2 to afford the phenolate-bridged species [(L2)M2(mu-X)X2] (M = Fe, X = Cl (2a); M = Co, X = Cl (2b); M = Ni, X = Br (2c); M = Zn, X = Cl (2d)). 1H NMR studies of diamagnetic 1d and 2d reveal that the limited rotation of the N aryl groups in 1d is further impeded in 2d by steric interactions imparted by the two closely located N-aryl groups. Partial displacement of the bridging bromide in 2c results upon its treatment with acetonitrile to afford [(L2)Ni2Br3(NCMe)] [2c(MeCN)]; no such reaction occurs for 2a, 2b, or 2d. Upon activation with excess methylalumoxane (MAO), 1b, 1c, 2b, and 2c show some activity for alkene oligomerization forming low molecular-weight materials with methyl-branched products predominating for the nickel systems. Single-crystal X-ray diffraction studies have been performed on L2-H, 1c, 2b, 2c, 2c(NCMe), and 2d. PMID- 17112288 TI - Absorption, light emission, and upconversion properties of Tm2+-doped CsCaI3 and RbCaI3. AB - Absorption, light emission, and upconversion properties of Tm2+-doped CsCaI3 and RbCaI3 single crystals are presented and compared. Both compounds show multiple emissions after excitation at 21834 cm(-1) between 10 and 300 K. Besides sharp 4f 4f transitions around 8800 cm(-1), five and three broad 4f-5d emission bands are observed at higher energies in CsCaI3:Tm2+ and RbCaI3:Tm2+, respectively. The 4f 5d transitions are very sensitive to the crystalline environment: the onset of the 4f-5d excitations is red-shifted by about 1000 cm(-1) in RbCaI3:Tm2+ compared to CsCaI3:Tm2+. In addition, a broadening of bands is observed in the former compound. These differences are attributed to the structural changes that occur when the alkali metal is changed from Cs to Rb in these crystal lattices. An increased energy splitting of the multiplets and a red shift of the barycenter of the (4f)12(5d)1 electron configuration in RbCaI3:Tm2+ is the result. This affects not only the color of the visible emission, which turns from green in CsCaI3:Tm2+ to yellow in RbCaI3:Tm2+, but also the excited state dynamics. As a consequence, the dominant upconversion processes are different in the two compounds. Thus, the two title compounds nicely illustrate the influence of the structural environment on the optical spectroscopic properties of Tm2+. PMID- 17112289 TI - Lanthanide and uranium complexes with an SPS-based pincer ligand. AB - Reactions of Ln(BH4)3(THF)n and [Li(Et2O)]SPS(Me)], the lithium salt of an anionic SPS pincer ligand composed of a central hypervalent lambda4-phosphinine ring bearing two ortho-positioned diphenylphosphine sulfide sidearms, led to the monosubstituted compounds [Ln(BH4)2(SPS(Me))(THF)2] [Ln = Ce (1), Nd (2)], while the homoleptic complexes [Ln(SPS(Me))3] [Ln = Ce (3), Nd (4)] were obtained by treatment of LnX3 (X = I, BH4) with [K(Et2O)][SPS(Me)]. The [UX2(SPS(Me))2] complexes [X = Cl (5), BH4 (6)] were isolated from reactions of UX4 and the lithium or potassium salt of the [SPS(Me)]- anion. The X-ray crystal structures of 1.1.5THF, 2.1.5THF, 3.2THF.2Et2O, and 5.4py reveal that the flexible tridentate [SPS(Me)]- anion is bound to the metal as a tertiary phosphine with electronic delocalization within the unsaturated parts of the ligand. PMID- 17112290 TI - Mono{hydrotris(mercaptoimidazolyl)borato} complexes of manganese(II), iron(II), cobalt(II), and nickel(II) halides. AB - A series of [Tm(Me)M(mu-Cl)]2 and Tm(R)MCl (Tm(R) = tris(mercaptoimidazolyl)borate; R = Me, tBu, Ph, 2,6-iPr2C6H3 (Ar); M = Mn, Fe, Co, Ni) complexes have been prepared by treatment of NaTm(Me) or LiTm(R) with an excess amount of metal(II) chlorides, MCl2. Treatment of Tm(R)MCl (R = tBu, Ph, Ar) with NaI led to a halide exchange to afford Tm(R)MI. The molecular structures of [Tm(Me)M(mu-Cl)]2 (M = Mn, Ni), [Tm(Me)Ni(mu-Br)]2, Tm(tBu)MCl (M = Fe, Co), Tm(Ph)MCl (M = Mn, Fe, Co, Ni), Tm(Ar)MCl (M = Mn, Fe, Co, Ni), Tm(Ph)MI (M = Mn, Co), and Tm(Ar)MI (M = Fe, Co, Ni) have been determined by X-ray crystallography. The Tm(R) ligands occupy the tripodal coordination site of the metal ions, giving a square pyramidal or trigonal bipyramidal coordination geometry for Tm(Me)M(mu Cl)]2 and a tetrahedral geometry for the Tm(R)MCl complexes, where the S-M-S bite angles are larger than the reported N-M-N angles of the corresponding hydrotris(pyrazolyl)borate (Tp(R)) complexes. Treatment of Tm(Ph)2Fe with excess FeCl2 affords Tm(Ph)FeCl, indicating that Tm(R)2M as well as Tm(R)MCl is formed at the initial stage of the reaction between MCl2 and the Tm(R) anion. PMID- 17112293 TI - Pharmacology of drugs formulated with DepoFoam: a sustained release drug delivery system for parenteral administration using multivesicular liposome technology. AB - Lamellar liposome technology has been used for several decades to produce sustained-release drug formulations for parenteral administration. Multivesicular liposomes are structurally distinct from lamellar liposomes and consist of an aggregation of hundreds of water-filled polyhedral compartments separated by bi layered lipid septa. The unique architecture of multivesicular liposomes allows encapsulating drug with greater efficiency, provides robust structural stability and ensures reliable, steady and prolonged drug release. The favourable characteristics of multivesicular liposomes have resulted in many drug formulations exploiting this technology, which is proprietary and referred to as DepoFoam. Currently, two formulations using multivesicular liposome technology are approved by the US FDA for clinical use, and many more formulations are at an experimental developmental stage. The first clinically available formulation contains the antineoplastic agent cytarabine (DepoCyt) for its intrathecal injection in the treatment of malignant lymphomatous meningitis. Intrathecal injection of DepoCyt reliably results in the sustained release of cytarabine and produces cytotoxic concentrations in cerebrospinal fluid (CSF) that are maintained for at least 2 weeks. Early efficacy data suggest that DepoCyt is fairly well tolerated, and its use allows reduced dosing frequency from twice a week to once every other week and may improve the outcome compared with frequent intrathecal injections of unencapsulated cytarabine. The second available formulation contains morphine (DepoDur) for its single epidural injection in the treatment of postoperative pain. While animal studies confirm that epidural injection of DepoDur results in the sustained release of morphine into CSF, the CSF pharmacokinetics have not been determined in humans. Clinical studies suggest that the use of DepoDur decreases the amount of systemically administered analgesics needed for adequate postoperative pain control. It may also provide superior pain control during the first 1-2 postoperative days compared with epidural administration of unencapsulated morphine or intravenous administration of an opioid. However, at this timepoint the overall clinical utility of DepoDur has yet to be defined and some safety concerns remain because of the unknown CSF pharmacokinetics of DepoDur in humans. The versatility of multivesicular liposome technology is reflected by the many agents including small inorganic and organic molecules and macromolecules including proteins that have successfully been encapsulated. Data concerning many experimental formulations containing antineoplastic, antibacterial and antiviral agents underscore the sustained, steady and reliable release of these compounds from multivesicular liposomes after injection by the intrathecal, subcutaneous, intramuscular, intraperitoneal and intraocular routes. Contingent on the specific formulation and manufacturing process, agents were released over a period of hours to weeks as reflected by a 2 to 400-fold increase in elimination half life. Published data further suggest that the encapsulation process preserves bioactivity of agents as delicate as proteins and supports the view that examined multivesicular liposomes were non toxic at studied doses. The task ahead will be to examine whether the beneficial structural and pharmacokinetic properties of multivesicular liposome formulations will translate into improved clinical outcomes, either because of decreased drug toxicity or increased drug efficacy. PMID- 17112294 TI - Pharmacokinetic profile of dexloxiglumide. AB - Dexloxiglumide is a potent and selective cholecystokinin type 1 (CCK1) receptor antagonist currently under development in a variety of diseases affecting the gastrointestinal tract such as gastro-oesophageal reflux disease, irritable bowel syndrome (IBS), functional dyspepsia, constipation and gastric emptying disorders. In female patients with constipation-predominant IBS, clinical efficacy has been demonstrated following administration of dexloxiglumide 200 mg three times daily. Dexloxiglumide is rapidly and extensively absorbed after single oral administration in humans with an absolute bioavailability of 48%. The incomplete bioavailability is due to both incomplete absorption and hepatic first pass effect. Following multiple-dose administration of 200 mg three times daily, the accumulation is predictable, indicating time-independent pharmacokinetics. In addition, dexloxiglumide pharmacokinetics are dose-independent after both single and repeated oral three-times-daily doses in the dose range 100-400 mg. Dexloxiglumide absorption window extends from the jejunum to the colon and the drug is a substrate and a weak inhibitor of P-glycoprotein and multidrug resistance protein 1. Plasma protein binding of dexloxiglumide is 94-98% and the drug has a moderate to low volume of distribution in humans. Systemic clearance of dexloxiglumide is moderate and cytochrome P450 (CYP) 3A4/5 and CYP2C9 have been implicated in the metabolism of dexloxiglumide to produce O-demethyl dexloxi glumide. This metabolite is further oxidised to dexloxiglumide carboxylic acid. These two major metabolites (accounting for up to 50% of dexloxiglumide elimination) have been identified. However, in human plasma the unchanged drug represents the major (up to 91%) component of the metabolic profile. The parent drug is believed to be the major contributor to the efficacy of the compound, since its major metabolites are pharmacologically inactive. In addition, the drug is a single isomer chiral drug (eutomer) that does not undergo chiral inversion into its pharmacologically inactive enantiomer (distomer). After oral administration of (14)C-dexloxiglumide, radioactivity is mainly excreted in bile and in faeces (74% of dose) with much lower excretion in urine (20% of dose). Renal excretion of unchanged dexloxiglumide is low (7% of dose in urine and faeces, 1% of dose in urine) and is dose-independent in the dose range 100-400 mg. As the kidney is a minor contributor to the elimination of dexloxiglumide and/or its metabolites in humans, the pharmacokinetics of the drug should not be affected in patients with renal insufficiency. The pharmacokinetics of dexloxiglumide are also not affected by age, sex and administration with a high fat breakfast. Mild and moderate liver impairment do not affect the pharmacokinetics of dexloxiglumide but severe liver impairment causes increases in systemic exposure to dexloxiglumide and O-demethyl dexloxiglumide. Thus, the drug should be prescribed with caution in patients with severe hepatic impairment even though no dose adjustment is warranted. The results of different drug interaction studies have indicated that no clinically relevant metabolic and concomitant drug-drug interactions are expected during the clinical use of dexloxiglumide. PMID- 17112295 TI - Pharmacogenetics of target genes across the warfarin pharmacological pathway. AB - Warfarin is a widely prescribed anticoagulant for thromboembolic disorders and exhibits wide inter-individual differences in its pharmacodynamic effects. Warfarin exerts its anticoagulant effect by inhibiting the enzymatic activity of vitamin K 2,3-epoxide reductase complex, subunit 1 (VKORC1) which regenerates reduced vitamin K as an essential cofactor for the post-translational gamma carboxylation of glutamic acid residues on coagulation factors II, VII, IX and X, and the anticoagulant proteins C, S and Z. Recent studies have shown polymorphisms in genes involved in the uptake of vitamin K (apolipoprotein E [ApoE]), reduction of vitamin K 2,3-epoxide (VKORC1), metabolism of warfarin (cytochrome P450 2C9 [CYP2C9]), and gamma carboxylation (gamma-glutamyl carboxylase [GGCX]) to influence the pharmacokinetics and pharmacodynamics of warfarin in patients from different ethnic backgrounds, resulting in variable warfarin dose requirements. Understanding the causal relationship of these polygenic influences on warfarin dose requirements in patients of different ethnicity may be vital in reducing inter-patient variability and optimising anticoagulant therapy. PMID- 17112296 TI - A population pharmacokinetic/pharmacodynamic analysis of regadenoson, an adenosine A2A-receptor agonist, in healthy male volunteers. AB - OBJECTIVES: The aims of this study were to investigate the safety, tolerability, pharmacokinetics and pharmacodynamics of regadenoson (CVT-3146) in healthy, male volunteers. METHODS: Thirty-six healthy, male volunteers aged 18-50 years were included in this randomised, double-blind, crossover, placebo-controlled study to evaluate single intravenous bolus doses of regadenoson that ranged from 0.1 to 30.0 micro g/kg. Subjects received one dose of regadenoson or placebo on successive days while supine, then the same dose of regadenoson or placebo on successive days while standing. As part of the safety evaluation, vital signs and adverse events were monitored and recorded throughout the course of the study in all subjects. Up to 20 plasma samples were collected for regadenoson concentration determination within the 24 hours after each supine dosage. All urine was collected during the 24-hour time period post-dose and an aliquot was used for the determination of the regadenoson concentration. Heart rate and blood pressure were recorded at many of the same timepoints that the samples for the pharmacokinetic analysis were taken. A non linear mixed-effect modelling approach, using the software NONMEM, was utilised in modelling the plasma and urine concentration-time profiles and temporal changes in heart rate after regadenoson administration in the supine position. The influences of several covariates, including bodyweight, body mass index and age, on pharmacokinetic model parameters were investigated. RESULTS: Adverse events were more prevalent at regadenoson doses above 3 micro g/kg, and the increase in the occurrence of adverse events was dose-related. Most of the adverse events were related to vasodilation and an increase in heart rate and were generally of mild to moderate severity. Based on the severity and frequency of adverse events, the maximum tolerated doses of regadenoson were deemed to be 10 micro g/kg in the standing position and 20 micro g/kg in the supine position. The pharmacokinetics of regadenoson were successfully described by a three-compartment model with linear clearance. Following intravenous bolus dose administration, regadenoson was rapidly distributed throughout the body, followed by relatively slower elimination (terminal elimination half-life of approximately 2 hours). The clearance was estimated to be 37.8 L/h, with renal excretion accounting for approximately 58% of the total elimination. The volume of distribution of the central compartment and the volume of distribution at steady state were estimated to be 11.5L and 78.7L, respectively. Individual pharmacokinetic parameter estimates were fixed in the pharmacodynamic model, where changes in heart rate were related to plasma drug concentrations using a Michaelis-Menten model. The maximum heart rate increase (Emax) and plasma regadenoson concentration causing a 50% increase in the maximum heart rate (EC50) were estimated to be 76 beats per minute and 12.3 ng/mL, respectively. None of the tested covariates was found to be correlated with any of the pharmacokinetic model parameters. CONCLUSIONS: The pharmacokinetics and the effects of regadenoson on heart rate were successfully described using pharmacokinetic/pharmacodynamic modelling. The lack of a correlation between the model estimates and various baseline patient demographics supports unit-based dose administration of regadenoson. PMID- 17112297 TI - Predicting pharmacokinetic food effects using biorelevant solubility media and physiologically based modelling. AB - BACKGROUND: Food-induced changes in gastric emptying time, gastric pH and/or intestinal fluid composition may have an impact on the pharmacokinetics of drugs. The aim of this work was to use mathematical models describing physiology in fed and fasted states together with biorelevant solubility and degradation data to simulate food effects for six compounds from recent Roche projects. METHODS: The solubility of each compound was measured in different biorelevant media: simulated human gastric fluid for the fasted and fed state, simulated human intestinal fluid for the fasted, fed and high-fat state, and simulated human colonic fluid for the upper and the lower colon. A physiologically based absorption model was developed in GastroPlustrade mark for each compound using permeability, solubility, metabolism and distribution data. By incorporating the appropriate physiological parameters and solubility data into the model, the oral pharmacokinetics of each drug was simulated under fasted, fed and/or high-fat conditions. Predicted and observed plasma concentration-time profiles and food effects were compared for a range of doses to assess the accuracy of the simulations. RESULTS: The models were able to distinguish between minor and significant food effects. The simulation captured well the magnitude of the food effects and for the six compounds correctly predicted the observed plasma exposure in fasted, fed and high-fat conditions. CONCLUSION: Biorelevant solubility tests can be used together with physiologically based absorption models to predict clinical food effects caused by solubility and/or dissolution rate limitations. PMID- 17112298 TI - Population pharmacokinetics of high-dose methotrexate in children with acute lymphoblastic leukaemia. AB - OBJECTIVE: To develop and a priori validate a methotrexate population pharmacokinetic model in children with acute lymphoblastic leukaemia (ALL), receiving high-dose methotrexate followed by folinic acid rescue, identifying the covariates that could explain part of the pharmacokinetic variability of methotrexate. METHODS: The study was carried out in 49 children (aged 6 months to 17 years) who received high-dose methotrexate (3 g/m(2) per course) in long-term treatment. In an index group (37 individuals; 1236 methotrexate plasma concentrations), a population pharmacokinetic model was developed using a nonlinear mixed-effects model. The remaining patients' data (12 individuals; 278 methotrexate plasma concentrations) were used for model validation. Age, sex, total bodyweight (TBW), height, body surface area, lowest urine pH during infusion, serum creatinine, ALT, AST, folinic acid dose and length of rescue were analysed as possible covariates. The final predictive performance of the pharmacokinetic model was tested using standardised mean prediction errors. RESULTS: The final population pharmacokinetic model (two-compartmental) included only age and total bodyweight as influencing clearance (CL) and volume of distribution of central compartment (V(1)). For children aged < or =10 years: CL (L/h) = 0.287 . TBW(0.876); V(1) (L) = 0.465 . TBW, and for children aged >10 years: CL (L/h) = 0.149 . TBW; V(1) (L) = 0.437 . TBW. From the base to the final model, the inter-individual variabilities for CL and V(1) were significantly reduced in both age groups (30-50%). The coefficients of variation of the pharmacokinetic parameters were <30%, while residual and inter-occasional coefficients maintained values close to 40%. Validation of the proposed model revealed the suitability of the model. CONCLUSION: A methotrexate population pharmacokinetic model has been developed for ALL children. The proposed model could be used in Bayesian algorithms with a limited sampling strategy to estimate the systemic exposure of individual patients to methotrexate and adapt both folinic acid rescue and methotrexate dosing accordingly. PMID- 17112300 TI - Anti-tumour necrosis factor therapy for ulcerative colitis: evidence to date. AB - Infliximab, the chimeric monoclonal antibody directed against tumour necrosis factor (TNF)-alpha, has profoundly changed therapy for Crohn's disease (CD). However, for ulcerative colitis (UC), before the publication of ACT 1 and ACT 2 (Active Ulcerative Colitis Trials 1 and 2), there were only a few open-label and controlled trials that evaluated the role of infliximab in the treatment of UC. Results from these earlier studies were equivocal and ambiguous. However, the ACT 1 and ACT 2 trials were large, randomised and placebo-controlled, and have shown that infliximab is significantly more efficacious than placebo in treating both corticosteroid-responsive and -refractory moderate to severe UC. Data from these two studies showed that in patients with moderate to severe UC, treatment with infliximab (5 and 10 mg/kg), compared with placebo, led to significantly higher rates of clinical response, clinical remission and mucosal healing. However, a significant proportion of patients who were receiving oral corticosteroids at the start of the trials, remained on corticosteroids despite infliximab therapy. Additionally, the safety profile of the drug was found to be similar to what has been reported in clinical studies of infliximab in patients with CD. On the basis of currently available data, we use infliximab as a remission-inducing agent in patients who have moderate to severe UC and are either refractory to or intolerant of mesalazine (5-ASA) products and immunomodulators. Moreover, infliximab seems to be a reasonable therapeutic modality for remission maintenance in those patients with UC in whom mesalazine products and immunomodulators have failed. Although data are limited, infliximab may be considered as a remission-inducing agent in patients with moderate to severe UC which is refractory to oral corticosteroids. However, the role of infliximab in the treatment of UC patients who are dependent on oral corticosteroids is still unclear and, therefore, should be considered only in patients who cannot be successfully transitioned to or are intolerant of oral immunomodulators. Furthermore, infliximab may be an alternative to ciclosporin (cyclosporin) in hospitalised patients with severe to moderately severe but not fulminant UC who do not respond to intravenous corticosteroids. At present, there is insufficient evidence to advocate using infliximab as a first-line agent for UC patients with mild or moderate to severe disease. Future randomised, controlled trials with clearly defined patient populations should further help to clarify the definitive role of infliximab in the therapeutic scheme for UC. PMID- 17112299 TI - Biochemical markers of bone turnover in the clinical development of drugs for osteoporosis and metastatic bone disease: potential uses and pitfalls. AB - Biochemical markers of bone turnover are used increasingly during the clinical development of drugs for the treatment of metabolic bone diseases such as Paget's disease, osteoporosis and cancer that has metastasised to the bone. However, assessing the optimal value of these markers is often complicated, and such an assessment is an obvious prerequisite for rational use of the markers and, consequently, potential improvement of clinical drug development. Biochemical markers of bone turnover are substances in the blood or urine that are produced or released during bone remodelling. They provide semiquantitative information on bone remodelling, and are often the most adequate tool to describe the pharmacodynamics of the drug. Their use has increased considerably because of dose-effect relationships that have been seen with certain drugs, but also because they have proven relationships with clinical outcomes in several metabolic bone diseases. However, there is a lack of information on the kinetics of these markers, and the immunoassays that are frequently used in their monitoring often measure a mixture of fragments rather than a single molecular entity. For drug development it should also be realised that different markers, but also different assays for the same marker, may provide different results, considerably limiting the ability to compare results. In postmenopausal osteoporosis, relationships have been shown between several biochemical markers of bone turnover, and either fracture risk and/or the antifracture efficacy of drugs. Such relationships can be used for the development of drugs with similar mechanisms of action, but also for the development of these drugs for closely related indications, such as corticosteroid-induced osteoporosis. In both of these instances, data on effects on biochemical markers of bone turnover are usually employed in combination with information about effects on bone mineral density. However, the relationships of these parameters with clinical outcomes may be remarkably different for drugs with alternative mechanisms of action, challenging the use of the markers for the development of new drugs for the treatment of patients with osteoporosis. At present, the pharmacological treatment of cancer that has metastasised to the bone is limited to several bisphosphonates. Recent studies have shown relationships between the normalisation of levels of biochemical markers of bone turnover and clinical outcomes, and prospective studies investigating the application of such relationships are ongoing. The markers may play an important role in the optimisation of registered bisphosphonate treatments. However, their role in the development of new drugs is still limited to dose selection, and potential relationships with clinical outcomes remain to be investigated in instances of new mechanisms of action. Biochemical markers of bone turnover are a valuable asset for drug development, but their rational use is determined by a number of variables. Correctly manipulating these may improve clinical development of drugs for the treatment of patients with metabolic bone diseases such as osteoporosis and cancer metastatic to the bone. PMID- 17112301 TI - Pilsicainide for atrial fibrillation. AB - Pilsicainide is a class IC antiarrhythmic drug, which has a pure sodium channel blocking action with slow recovery pharmacokinetics. In experimental studies, pilsicainide has a depressant effect on intra-atrial conduction and a prolonging effect on the atrial effective refractory period (ERP). In patients with paroxysmal atrial fibrillation (AF), pilsicainide significantly prolonged the ERP of the distal pulmonary vein (PV), PV-left atrium (LA) junction and LA, and the conduction time from the distal PV to the PV-LA junction. In some patients, PV-LA conduction block has been observed just before pilsicainide-induced termination of AF; this isolation of the PV may provide a new insight into the mechanism of pharmacological conversion of AF. Hybrid therapy with pilsicainide and PV isolation (by radiofrequency catheter ablation) appears to be an effective therapeutic approach for AF. The pharmacological PV isolation by pilsicainide and its suppression of focal discharges from atrial tissue may prevent the development of AF after unsuccessful ablation. PMID- 17112303 TI - Treatment of depression in acute coronary syndromes with selective serotonin reuptake inhibitors. AB - Depression in patients with acute coronary syndromes (ACS) is common and associated with impaired cardiovascular prognosis in terms of cardiac mortality and new cardiovascular events. It remains unclear whether antidepressant treatment may reverse these effects. In this review, the literature is evaluated on (i) the antidepressant efficacy of selective serotonin reuptake inhibitors (SSRIs) for depression in patients with ACS; (ii) the pleiomorphic effects of SSRIs that may be associated with cardiovascular prognosis; and (iii) the effects of SSRIs on cardiovascular prognosis.SSRIs provide modest relief of depressive symptoms in selected subgroups of depressed patients with ACS. With respect to the pleiomorphic effects of SSRIs, three mechanisms of how SSRIs may improve cardiovascular prognosis are discussed: via platelet function, via the autonomic nervous system (ANS) and via vasomotor tone. Some studies show that SSRIs may reduce platelet activity and sympathetic nervous system activation, but results are inconclusive. SSRIs are associated with vasodilation but this needs to be confirmed with in vivo experiments. Some non-experimental studies describe favourable effects of SSRIs on cardiovascular prognosis. Despite recent developments, much of the effect of SSRIs on cardiovascular prognosis remains unclear. Although some studies suggest effects of SSRIs on platelet function, ANS and vasomotor tone, which may lead to improved cardiovascular prognosis, results are largely inconclusive. More well designed studies addressing these questions are needed. Moreover, since the effects of SSRIs on depression itself are limited, efforts should be dedicated to study the diagnostic validity and homogeneity of depression in the context of ACS and the presence of clinically relevant subtypes. PMID- 17112304 TI - Rimonabant. AB - Rimonabant is the first of a new class of selective cannabinoid receptor-1 blockers. It reduces the overactivity of the endocannabinoid system, improving lipid and glucose metabolism and regulating food intake and energy balance. In four randomised, double-blind clinical trials in overweight or obese adults with or without type 2 diabetes and/or dyslipidaemia, oral rimonabant 20mg once daily reduced weight and waist circumference to a significantly greater extent than placebo. A significantly greater proportion of rimonabant than placebo recipients achieved the clinically significant weight-loss target of > or =5% or > or =10% of initial weight. Rimonabant was associated with significant improvements in glycaemic control relative to placebo, with approximately equal to 57% of the reduction in glycosylated haemoglobin being independent of the effects of weight loss in one trial. Improvements in other cardiometabolic risk factors (i.e. increases in high-density lipoprotein-cholesterol [HDL-C] and decreases in triglyceride [TG] levels) were significantly greater with rimonabant than with placebo. The improvement in lipid profile also demonstrated a weight-independent effect, with approximately equal to 47-58% of the improvement in HDL-C and TG being beyond that expected through weight loss alone. Rimonabant was generally well tolerated, with most adverse events considered mild to moderate in severity. PMID- 17112302 TI - Disease-modifying therapies in Alzheimer's disease: how far have we come? AB - Currently, there are no disease-modifying therapies available for Alzheimer's disease (AD). Acetylcholinesterase inhibitors and memantine are licensed for AD and have moderate symptomatic benefits. Epidemiological studies have suggested that NSAIDs, estrogen, HMG-CoA reductase inhibitors (statins) or tocopherol (vitamin E) can prevent AD. However, prospective, randomised studies have not convincingly been able to demonstrate clinical efficacy. Major progress in molecular medicine suggests further drug targets. The metabolism of the amyloid precursor protein and the aggregation of its Abeta fragment are the focus of current studies. Abeta peptides are produced by the enzymes beta- and gamma secretase. Inhibition of gamma-secretase has been shown to reduce Abeta production. However, gamma-secretase activity is also involved in other vital physiological pathways. Involvement of gamma-secretase in cell differentiation may preclude complete blockade of gamma-secretase for prolonged times in vivo. Inhibition of beta-secretase seems to be devoid of serious adverse effects according to studies with knockout animals. However, targeting beta-secretase is hampered by the lack of suitable inhibitors to date. Other approaches focus on enzymes that cut inside the Abeta sequence such as alpha-secretase and neprilysin. Stimulation of the expression or activity of alpha-secretase or neprilysin has been shown to enhance Abeta degradation. Furthermore, inhibitors of Abeta aggregation have been described and clinical trials have been initiated. Peroxisome proliferator activated receptor-gamma agonists and selected NSAIDs may be suitable to modulate both Abeta production and inflammatory activation. On the basis of autopsy reports, active immunisation against Abeta in humans seems to have proven its ability to clear amyloid deposits from the brain. However, a first clinical trial with active vaccination against the full length Abeta peptide has been halted because of adverse effects. Further trials with vaccination or passive transfer of antibodies are planned. PMID- 17112308 TI - Persistent, scaly facial erythema with erosions. Hydrocortisone and shampooing offer no relief. PMID- 17112307 TI - Intravenous droperidol: a review of its use in the management of postoperative nausea and vomiting. AB - Droperidol (Dehydrobenzperidol, Dehidrobenzoperidol, Dridol, Droleptan, Inapsine) is a dopamine D(2) receptor antagonist that has been widely used in adults and children for the prevention and treatment of postoperative nausea and vomiting (PONV) over several decades and, more recently, for the prevention of opioid induced PONV during patient-controlled analgesia (PCA) in adults. In well controlled clinical trials of patients undergoing surgery, the efficacy of single dose intravenous (IV) droperidol in preventing PONV was similar to that of ondansetron and dexamethasone. Droperidol significantly reduced opioid-induced PONV in adults during PCA and had a morphine-sparing effect. Droperidol is generally well tolerated and the incidence of adverse effects is similar to that observed with placebo and the serotonin 5-HT(3) receptor antagonists (setrons). Guidelines recommend that, in adults, droperidol monotherapy be considered for those at moderate risk of PONV, and droperidol in combination with a setron and/or dexamethasone be considered for patients at moderate or high risk of PONV. In children with moderate or high risk of PONV, droperidol is recommended for first-line use in some countries, and second-line use in others. PMID- 17112309 TI - Grapefruit juice and drug interactions. Exploring mechanisms of this interaction and potential toxicity for certain drugs. AB - Concomitant administration of grapefruit juice can increase the plasma concentration of numerous drugs in humans and decrease the concentration of a few others. Such elevations of drug plasma concentrations have, on occasion, resulted in adverse clinical effects. Increased concentrations are primarily mediated by chemicals in grapefruit juice, which inhibit the CYP 3A4 drug-metabolizing enzyme in the small intestines. This inhibition decreases the first-pass metabolism of drugs using the CYP 3A4 intestinal system and increases the bioavailability and maximal plasma drug concentrations (Cmax) of the CYP 3A4 substrates. The effect of grapefruit juice on drug metabolism is most pronounced in drugs with a high first-pass metabolism (eg, felodipine, amiodarone), in which it inhibits the first-pass metabolism of the CYP 3A4 substrates leading to an increase in Cmax and area under the concentration time curve (AUC). The use of grapefruit juice with a few specific drugs (eg, fexofenadine, digoxin) may lower plasma drug concentrations by inhibiting drug absorption catalyzed by the organic anion transporting polypeptide (OATP). PMID- 17112310 TI - Alcohol use in the elderly: beyond the CAGE. Part 2: Screening instruments and treatment strategies. AB - Recognition of the prevalence of alcohol problems in older patients has led researchers to develop and standardize screening instruments specifically intended for use in the elderly. While the CAGE questionnaire is the most commonly used screening instrument for alcohol use disorders, its usefulness in the geriatric patient has been questioned. Other tools, such as the Alcohol Use Disorders Identification Test (AUDIT), Michigan Alcohol Screening Test-Geriatric Version (MAST-G), and the Alcohol-Related Problems Survey (ARPS) may have additional use in this population. Brief interventions have been shown to be effective in producing sustained abstinence or reducing levels of consumption, thereby decreasing hazardous and harmful drinking. Clinicians may identify specific changes in the medical management of the individual that will reduce risk regardless of drinking behavior. PMID- 17112311 TI - Funneled landscape leads to robustness of cell networks: yeast cell cycle. AB - We uncovered the underlying energy landscape for a cellular network. We discovered that the energy landscape of the yeast cell-cycle network is funneled towards the global minimum (G0/G1 phase) from the experimentally measured or inferred inherent chemical reaction rates. The funneled landscape is quite robust against random perturbations. This naturally explains robustness from a physical point of view. The ratio of slope versus roughness of the landscape becomes a quantitative measure of robustness of the network. The funneled landscape can be seen as a possible realization of the Darwinian principle of natural selection at the cellular network level. It provides an optimal criterion for network connections and design. Our approach is general and can be applied to other cellular networks. PMID- 17112312 TI - Optimal noise filtering in the chemotactic response of Escherichia coli. AB - Information-carrying signals in the real world are often obscured by noise. A challenge for any system is to filter the signal from the corrupting noise. This task is particularly acute for the signal transduction network that mediates bacterial chemotaxis, because the signals are subtle, the noise arising from stochastic fluctuations is substantial, and the system is effectively acting as a differentiator which amplifies noise. Here, we investigated the filtering properties of this biological system. Through simulation, we first show that the cutoff frequency has a dramatic effect on the chemotactic efficiency of the cell. Then, using a mathematical model to describe the signal, noise, and system, we formulated and solved an optimal filtering problem to determine the cutoff frequency that bests separates the low-frequency signal from the high-frequency noise. There was good agreement between the theory, simulations, and published experimental data. Finally, we propose that an elegant implementation of the optimal filter in combination with a differentiator can be achieved via an integral control system. This paper furnishes a simple quantitative framework for interpreting many of the key notions about bacterial chemotaxis, and, more generally, it highlights the constraints on biological systems imposed by noise. PMID- 17112313 TI - 3D complex: a structural classification of protein complexes. AB - Most of the proteins in a cell assemble into complexes to carry out their function. It is therefore crucial to understand the physicochemical properties as well as the evolution of interactions between proteins. The Protein Data Bank represents an important source of information for such studies, because more than half of the structures are homo- or heteromeric protein complexes. Here we propose the first hierarchical classification of whole protein complexes of known 3-D structure, based on representing their fundamental structural features as a graph. This classification provides the first overview of all the complexes in the Protein Data Bank and allows nonredundant sets to be derived at different levels of detail. This reveals that between one-half and two-thirds of known structures are multimeric, depending on the level of redundancy accepted. We also analyse the structures in terms of the topological arrangement of their subunits and find that they form a small number of arrangements compared with all theoretically possible ones. This is because most complexes contain four subunits or less, and the large majority are homomeric. In addition, there is a strong tendency for symmetry in complexes, even for heteromeric complexes. Finally, through comparison of Biological Units in the Protein Data Bank with the Protein Quaternary Structure database, we identified many possible errors in quaternary structure assignments. Our classification, available as a database and Web server at http://www.3Dcomplex.org, will be a starting point for future work aimed at understanding the structure and evolution of protein complexes. PMID- 17112314 TI - An integrative genomic approach to uncover molecular mechanisms of prokaryotic traits. AB - With mounting availability of genomic and phenotypic databases, data integration and mining become increasingly challenging. While efforts have been put forward to analyze prokaryotic phenotypes, current computational technologies either lack high throughput capacity for genomic scale analysis, or are limited in their capability to integrate and mine data across different scales of biology. Consequently, simultaneous analysis of associations among genomes, phenotypes, and gene functions is prohibited. Here, we developed a high throughput computational approach, and demonstrated for the first time the feasibility of integrating large quantities of prokaryotic phenotypes along with genomic datasets for mining across multiple scales of biology (protein domains, pathways, molecular functions, and cellular processes). Applying this method over 59 fully sequenced prokaryotic species, we identified genetic basis and molecular mechanisms underlying the phenotypes in bacteria. We identified 3,711 significant correlations between 1,499 distinct Pfam and 63 phenotypes, with 2,650 correlations and 1,061 anti-correlations. Manual evaluation of a random sample of these significant correlations showed a minimal precision of 30% (95% confidence interval: 20%-42%; n = 50). We stratified the most significant 478 predictions and subjected 100 to manual evaluation, of which 60 were corroborated in the literature. We furthermore unveiled 10 significant correlations between phenotypes and KEGG pathways, eight of which were corroborated in the evaluation, and 309 significant correlations between phenotypes and 166 GO concepts evaluated using a random sample (minimal precision = 72%; 95% confidence interval: 60%-80%; n = 50). Additionally, we conducted a novel large-scale phenomic visualization analysis to provide insight into the modular nature of common molecular mechanisms spanning multiple biological scales and reused by related phenotypes (metaphenotypes). We propose that this method elucidates which classes of molecular mechanisms are associated with phenotypes or metaphenotypes and holds promise in facilitating a computable systems biology approach to genomic and biomedical research. PMID- 17112315 TI - Epitope mapping and topographic analysis of VAR2CSA DBL3X involved in P. falciparum placental sequestration. AB - Pregnancy-associated malaria is a major health problem, which mainly affects primigravidae living in malaria endemic areas. The syndrome is precipitated by accumulation of infected erythrocytes in placental tissue through an interaction between chondroitin sulphate A on syncytiotrophoblasts and a parasite-encoded protein on the surface of infected erythrocytes, believed to be VAR2CSA. VAR2CSA is a polymorphic protein of approximately 3,000 amino acids forming six Duffy binding-like (DBL) domains. For vaccine development it is important to define the antigenic targets for protective antibodies and to characterize the consequences of sequence variation. In this study, we used a combination of in silico tools, peptide arrays, and structural modeling to show that sequence variation mainly occurs in regions under strong diversifying selection, predicted to form flexible loops. These regions are the main targets of naturally acquired immunoglobulin gamma and accessible for antibodies reacting with native VAR2CSA on infected erythrocytes. Interestingly, surface reactive anti-VAR2CSA antibodies also target a conserved DBL3X region predicted to form an alpha-helix. Finally, we could identify DBL3X sequence motifs that were more likely to occur in parasites isolated from primi- and multigravidae, respectively. These findings strengthen the vaccine candidacy of VAR2CSA and will be important for choosing epitopes and variants of DBL3X to be included in a vaccine protecting women against pregnancy associated malaria. PMID- 17112316 TI - Virulence attributes and hyphal growth of C. neoformans are quantitative traits and the MATalpha allele enhances filamentation. AB - Cryptococcus neoformans is a fungal human pathogen with a bipolar mating system. It undergoes a dimorphic transition from a unicellular yeast to hyphal filamentous growth during mating and monokaryotic fruiting. The traditional sexual cycle that leads to the production of infectious basidiospores involves cells of both alpha and a mating type. Monokaryotic fruiting is a modified form of sexual reproduction that involves cells of the same mating type, most commonly alpha, which is the predominant mating type in both the environment and clinical isolates. However, some a isolates can also undergo monokaryotic fruiting. To determine whether mating type and other genetic loci contribute to the differences in fruiting observed between alpha and a cells, we applied quantitative trait loci (QTL) mapping to an inbred population of F2 progeny. We discovered that variation in hyphal length produced during fruiting is a quantitative trait resulting from the combined effects of multiple genetic loci, including the mating type (MAT) locus. Importantly, the alpha allele of the MAT locus enhanced hyphal growth compared with the a allele. Other virulence traits, including melanization and growth at 39 degrees C, also are quantitative traits that share a common QTL with hyphal growth. The Mac1 transcription factor, encoded in this common QTL, regulates copper homeostasis. MAC1 allelic differences contribute to phenotypic variation, and mac1Delta mutants exhibit defects in filamentation, melanin production, and high temperature growth. Further characterization of these QTL regions will reveal additional quantitative trait genes controlling biological processes central to fungal development and pathogenicity. PMID- 17112317 TI - Defining synphenotype groups in Xenopus tropicalis by use of antisense morpholino oligonucleotides. AB - To identify novel genes involved in early development, and as proof-of-principle of a large-scale reverse genetics approach in a vertebrate embryo, we have carried out an antisense morpholino oligonucleotide (MO) screen in Xenopus tropicalis, in the course of which we have targeted 202 genes expressed during gastrula stages. MOs were designed to complement sequence between -80 and +25 bases of the initiating AUG codons of the target mRNAs, and the specificities of many were tested by (i) designing different non-overlapping MOs directed against the same mRNA, (ii) injecting MOs differing in five bases, and (iii) performing "rescue" experiments. About 65% of the MOs caused X. tropicalis embryos to develop abnormally (59% of those targeted against novel genes), and we have divided the genes into "synphenotype groups," members of which cause similar loss of-function phenotypes and that may function in the same developmental pathways. Analysis of the expression patterns of the 202 genes indicates that members of a synphenotype group are not necessarily members of the same synexpression group. This screen provides new insights into early vertebrate development and paves the way for a more comprehensive MO-based analysis of gene function in X. tropicalis. PMID- 17112318 TI - Four linked genes participate in controlling sporulation efficiency in budding yeast. AB - Quantitative traits are conditioned by several genetic determinants. Since such genes influence many important complex traits in various organisms, the identification of quantitative trait loci (QTLs) is of major interest, but still encounters serious difficulties. We detected four linked genes within one QTL, which participate in controlling sporulation efficiency in Saccharomyces cerevisiae. Following the identification of single nucleotide polymorphisms by comparing the sequences of 145 genes between the parental strains SK1 and S288c, we analyzed the segregating progeny of the cross between them. Through reciprocal hemizygosity analysis, four genes, RAS2, PMS1, SWS2, and FKH2, located in a region of 60 kilobases on Chromosome 14, were found to be associated with sporulation efficiency. Three of the four "high" sporulation alleles are derived from the "low" sporulating strain. Two of these sporulation-related genes were verified through allele replacements. For RAS2, the causative variation was suggested to be a single nucleotide difference in the upstream region of the gene. This quantitative trait nucleotide accounts for sporulation variability among a set of ten closely related winery yeast strains. Our results provide a detailed view of genetic complexity in one "QTL region" that controls a quantitative trait and reports a single nucleotide polymorphism-trait association in wild strains. Moreover, these findings have implications on QTL identification in higher eukaryotes. PMID- 17112319 TI - Functional identification of Api5 as a suppressor of E2F-dependent apoptosis in vivo. AB - Retinoblastoma protein and E2-promoter binding factor (E2F) family members are important regulators of G1-S phase progression. Deregulated E2F also sensitizes cells to apoptosis, but this aspect of E2F function is poorly understood. Studies of E2F-induced apoptosis have mostly been carried out in tissue culture cells, and the analysis of the factors that are important for this process has been restricted to the testing of a few candidate genes. Using Drosophila as a model system, we have generated tools that allow genetic modifiers of E2F-dependent apoptosis to be identified in vivo and developed assays that allow effects on E2F induced apoptosis to be studied in cultured cells. Genetic interactions show that dE2F1-dependent apoptosis in vivo involves dArk/Apaf1 apoptosome-dependent activation of both initiator and effector caspases and is sensitive to levels of Drosophila inhibitor of apoptosis-1 (dIAP1). Using these approaches, we report the surprising finding that apoptosis inhibitor-5/antiapoptosis clone-11 (Api5/Aac11) is a critical determinant of dE2F1-induced apoptosis in vivo and in vitro. This functional interaction occurs in multiple tissues, is specific to E2F induced apoptosis, and is conserved from flies to humans. Interestingly, Api5/Aac11 acts downstream of E2F and suppresses E2F-dependent apoptosis without generally blocking E2F-dependent transcription. Api5/Aac11 expression is often upregulated in tumor cells, particularly in metastatic cells. We find that depletion of Api5 is tumor cell lethal. The strong genetic interaction between E2F and Api5/Aac11 suggests that elevated levels of Api5 may be selected during tumorigenesis to allow cells with deregulated E2F activity to survive under suboptimal conditions. Therefore, inhibition of Api5 function might offer a possible mechanism for antitumor exploitation. PMID- 17112321 TI - Vulnerable populations: who are they? PMID- 17112322 TI - Benefit-based copays in the real world: the employer perspective. PMID- 17112323 TI - Impact of copays on vulnerable populations. PMID- 17112324 TI - Oral combination therapy with thiazolidinediones in type 2 diabetes. AB - Insulin resistance occurs early in the type 2 diabetes disease process, leading to progressive beta cell failure and overt diabetes. By the time the diagnosis of diabetes is made, advanced macrovascular disease may already be present. Monotherapy with a sulfonylurea or metformin can slow, but does not prevent, the progression of disease. Successful management requires combination therapy that addresses both insulin resistance and beta cell dysfunction. Clinical trials support the use of combinations of agents with complementary mechanisms of action, such as a sulfonylurea or metformin plus a thiazolidinedione. Early aggressive treatment can improve patient outcomes while reducing overall healthcare costs. PMID- 17112325 TI - Health outcomes beyond glucose control. AB - Direct and indirect costs for diabetes mellitus place a large economic burden on the US healthcare system. Diabetes and cardiovascular disease (CVD) are closely interrelated, and it is estimated that much of the burden of CVD too is attributable to diabetes. Insulin resistance may be the common link between diabetes and CVD, often manifested clinically as the metabolic syndrome. The thiazolidinediones (TZDs) reduce insulin resistance and have favorable effects on lipids, blood pressure, and other cardiovascular risk factors. Large clinical trials have shown that early, aggressive intervention with lifestyle changes and pharmacotherapy with TZDs and other agents may slow progression to overt diabetes in high-risk patients and reduce the risk of cardiovascular and other complications; this could reduce healthcare resource utilization and costs. PMID- 17112326 TI - Case study. PMID- 17112320 TI - Genomic variation in rice: genesis of highly polymorphic linkage blocks during domestication. AB - Genomic regions that are unusually divergent between closely related species or racial groups can be particularly informative about the process of speciation or the operation of natural selection. The two sequenced genomes of cultivated Asian rice, Oryza sativa, reveal that at least 6% of the genomes are unusually divergent. Sequencing of ten unlinked loci from the highly divergent regions consistently identified two highly divergent haplotypes with each locus in nearly complete linkage disequilibrium among 25 O. sativa cultivars and 35 lines from six wild species. The existence of two highly divergent haplotypes in high divergence regions in species from all geographical areas (Africa, Asia, and Oceania) was in contrast to the low polymorphism and low linkage disequilibrium that were observed in other parts of the genome, represented by ten reference loci. While several natural processes are likely to contribute to this pattern of genomic variation, domestication may have greatly exaggerated the trend. In this hypothesis, divergent haplotypes that were adapted to different geographical and ecological environments migrated along with humans during the development of domesticated varieties. If true, these high divergence regions of the genome would be enriched for loci that contribute to the enormous range of phenotypic variation observed among domesticated breeds. PMID- 17112327 TI - Ethnic, gender, and age-related differences in the treatment of dyslipidemia. AB - Coronary heart disease (CHD) remains the leading cause of mortality in the United States, and factors such as age, gender, or race/ethnicity have a significant impact on cardiovascular risk. More than 80% of people who die from CHD are 65 years or older. Because women experience myocardial infarction (MI) at older ages than men, their mortality from MI is greater than it is for men. The largest disparity can clearly be seen between white women and black women. CHD death rates are higher among blacks than whites and the discrepancy in rates of premature CHD death is even more pronounced. Given this information, aggressive treatment of CHD risk factors is critical in black individuals, older individuals, and women. Although abundant randomized, controlled clinical data exist to support the efficacy of lipid-lowering therapy in preventing CHD events, these populations have traditionally been underrepresented in intervention-based clinical trials, despite their high prevalence of CHD. This article will explore the evidence for instituting statin therapy as part of a risk reducing strategy in older individuals, women, and ethnic minorities. PMID- 17112328 TI - Differences between clinical trial efficacy and real-world effectiveness. AB - Aggressive lowering of low-density lipoprotein cholesterol (LDL-C) with statin therapy can reduce the incidence of morbidity and mortality from coronary heart disease (CHD) in primary and secondary prevention settings. Indeed, suboptimal statin treatment has been associated with an increased risk of CHD events. Surveys such as the Lipid Treatment Assessment Project (L-TAP) and National Cholesterol Education Program Evaluation Project Utilizing Novel E-Technology (NEPTUNE) II have demonstrated that patients in real-world clinical settings often fail to reach the target goals set forth by the National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III). This failure to reach target LDL-C levels in real-world clinical practice presents a therapeutic treatment gap. There may be multiple reasons for this discrepancy: lack of patient follow-up, absence of well-defined protocols (ie, use of low-potency statins in high-risk patients), adherence controls (eg, pill counts, refill records), cost, and lack of patient motivation are a few possibilities. Several large clinical trials since ATP III have shown that these goals are achievable through aggressive statin therapy. This review sets forth compelling data that starting patients on or switching to high-efficacy LDL-C-lowering therapy enhances achievement of NCEP ATP III guidelines outside of the controlled trial setting. PMID- 17112329 TI - Effectiveness and cost-effectiveness of rosuvastatin, atorvastatin, and simvastatin among high-risk patients in usual clinical practice. AB - BACKGROUND: Assessments of the effectiveness and cost-effectiveness of treatment with statins in high risk patients in routine clinical practice are needed. The objective of the present study was to estimate the clinical effectiveness and cost-effectiveness of rosuvastatin compared with atorvastatin or simvastatin among high-risk patients as treated in routine clinical practice. METHODS: Patients aged 18 to 79 years with coronary heart disease (CHD) or equivalent who initiated treatment with atorvastatin, rosuvastatin, or simvastatin were included. Primary outcome variables were the percent reduction in low-density lipoprotein cholesterol (LDL-C), achievement of the National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) LDL-C goal, treatment costs, and cost-effectiveness. Regression models were used to adjust outcome measures for age, sex, CHD, baseline LDL-C, and therapy duration. A decision analytic model was used to assess incremental cost-effectiveness. RESULTS: Of the 775 eligible patients, rosuvastatin patients had higher baseline LDL-C levels (156 mg/dL vs 142 mg/dL or 137 mg/dL, respectively) compared with atorvastatin or simvastatin. Adjusted for baseline factors, percent LDL-C reduction was significantly greater with rosuvastatin versus atorvastatin or simvastatin (37% vs 28% or 27%, respectively; P <.05). The estimated percentage of patients attaining NCEP ATP III goal was higher (P <.05) for rosuvastatin (69.7%) compared with atorvastatin (54.8%) or simvastatin (51.2%), adjusted for baseline characteristics. Rosuvastatin patients also had the lowest annualized treatment costs (934dollars vs 1050 dollars or 1545 dollars for atorvastatin or simvastatin). Rosuvastatin was more effective and less costly than atorvastatin and at current branded and generic prices of simvastatin. A 60% to 68% discount from simvastatin branded price was needed to achieve equivalent cost effectiveness as rosuvastatin. CONCLUSIONS: In clinical practice, rosuvastatin is more effective and cost-effective in lowering LDL-C and in attainment of ATP III LDL-C goals compared with atorvastatin or simvastatin among high-risk patients. PMID- 17112330 TI - Do audible and ultrasonic sounds of intensities common in animal facilities affect the autonomic nervous system of rodents? AB - In animal facilities, noises, often poorly controlled, occur over a wide range of frequencies and intensities. Evidence demonstrates that audible noise and ultrasound have deleterious effects on rodent physiology, but it is not known how they affect the autonomic nervous system (ANS). This study exposed 3 unrestrained, male, Sprague-Dawley rats daily to a 15-min white noise regime (90 dB), a quiet regime, or a 15-min ultrasound regime (90 dB at 4 frequencies in the range 20 to 40 kHz)--each for several weeks--and used radiotelemetry to monitor their cardiovascular responses. Exposure to audible noise increased heart rate and mean arterial pressure. Spectral analysis of HR variability showed diminished stimulation of the parasympathetic nervous system, shifting the sympathovagal balance. However, ultrasound, at the frequencies used, did not reproducibly affect cardiovascular parameters. The preliminary data obtained from this study indicate that audible noise, but not ultrasound (delivered using the same protocol), affects the ANS. Because the cardiovascular, respiratory, renal, and gastrointestinal systems are under autonomic control, such noise could have wide ranging effects on animal physiology. PMID- 17112331 TI - Community demographics and the propensity to report animal cruelty. AB - The last decade has seen an increased awareness concerning links between violence to nonhuman animals and violence to humans. This has resulted in a number of cross-reporting initiatives between family service providers and animal welfare organizations. The success of these initiatives rests on individuals being willing to report such violence. Thus, there is a need to determine which variables influence an individual's willingness to report deliberate animal cruelty and abuse. The aim of this study was to examine demographic and attitudinal variables to ascertain their impact on propensity to report deliberate animal harm. A telephone questionnaire resulted in 1,208 valid responses from members of the general community. Results showed a number of variables that affected the propensity to report: gender, occupation, and acknowledgment of the link between family violence and deliberate animal harm. This article discusses these variables and their implications. PMID- 17112332 TI - Measuring the heat loss in horses in different seasons by infrared thermography. AB - It is necessary to consider breed and cold tolerance in the housing and caring of horses. This study demonstrates differences in heat loss between horse types at low temperatures and examines rate of loss in different types during different seasons. Eighteen horses participated. Groups by type were light (L), warmblood (W), coldblood (C), and pony (P). A camera filmed thermographic images at 15 degrees C, 2 degrees C (all types), -8 degrees C (L, W, C), and -12 degrees C (P). The study calculated loss from the neck, trunk, and inner surfaces of front and hind legs. Loss was similar in all types at 15 degrees C. L, W, and C dissipated more heat at 2 degrees C than at 15 degrees C (p < .001) and from neck and trunk at -8 degrees C than at 2 degrees C (p < .05). P dissipated heat similarly at 2 degrees C and -12 degrees C. At 2 degrees C, loss was less from neck and trunk in C and P compared with L (p < .05). At -8 degrees C, loss in L and W was greater than in C (p < .05). PMID- 17112333 TI - Stereotypic behavior of a female Asiatic elephant (Elephas maximus) in a zoo. AB - This study recorded daytime behavior of a female Asiatic elephant at the Municipal Zoo, Wroclaw, Poland, in both an indoor pen and an outdoor paddock as continuous scan sampling for 140 hr, over 35 days in 1 year. Stereotypic sequences involved bouts of highly repetitive stereotypic movements and much more variable interbout behavior. The study found both stereotypic movements, nodding and body (corpus) swaying, were asymmetric, accompanied by protraction of the right hind leg and to-and-fro swinging of the trunk. The elephant spent 52% of the daytime in stereotypic movements, 3.5 times the level reported for females in other zoos' groups. The share of time devoted to stereotypic behavior was lowest in the summer when the elephant was regularly released to the paddock and highest in the late fall after she had stayed in the pen after months of days outside. This suggests that changes in the management routine enhance stereotypies. Comparing the summer and winter stable management periods, stereotypies were much more frequent in the indoor pen than the outdoor paddock, suggesting that the confinement to a barren pen contributed to the observed levels of stereotypies. PMID- 17112334 TI - Behavioral decisions for managing social distance and aggression in captive polar bears (Ursus maritimus). AB - The polar bear (Ursus maritimus) in most zoos attracts high levels of public attention and can play an important role in conservation education. Polar bears in the wild are typically solitary; bears in captivity often house socially. This study reported behavioral evidence on how bears manage this situation and whether proximity leads to aggression. The study recorded location and behavior once per minute for 106 hr for 2 female polar bears at the Philadelphia Zoo; the samples represented all times of day bears spent in the enclosure (off-exhibit time spent in separate, indoor dens). When changing locations, 1 bear more frequently moved away from the other, effecting a net increase in interindividual distance. When either bear moved into an adjacent zone, 1 typically moved away. The bears occupied the same enclosure zone for a low proportion of time; proximity did not routinely lead to overt aggression. These data indicate polar bears make behavioral decisions, minimizing aggression, to manage social distance and that enclosure designers for solitary species--to facilitate social avoidance--should consider using topographical complexity and multiple pathways throughout. PMID- 17112335 TI - A note on the effect of controlling stable flies (Stomoxys calcitrans) in the resting activity and pen distribution of dairy cows. AB - This study investigated the effect of controlling stable flies (Stomoxys calcitrans) on the number of dairy cows lying down and their pen distribution. The study randomly assigned 80 Holstein cows to 1 of 2 groups. The treated group (T) included cows individually sprayed with insecticide when found with an average of 10 stable flies per cow; in the control group (C), cows received no application of insecticide. The pen had 4 equal-size areas: (a) feeding, (b) drinking and sunny, (c) covered, and (d) manure. The study recorded the number of cows lying and the area of the pen where the nonhuman animal was located. The study found no difference (p > .05) between the proportion of T and C cows lying. However, cows preferred to lie down in pen area 3 when fewer than 10 stable flies per cow were found. Area 4 was the most avoided section of the pen, except when a high incidence of flies was present. The study concluded that high populations of S. calcitrans (> 10 flies per cow) did not affect the number of dairy cows lying down. However, it modified site preferences for lying. PMID- 17112336 TI - An hourly variation in zoo visitor interest: measurement and significance for animal welfare research. AB - A methodological difficulty facing welfare research on nonhuman animals in the zoo is the large number of uncontrolled variables due to variation within and between study sites. Zoo visitors act as uncontrolled variables, with number, density, size, and behavior constantly changing. This is worrisome because previous research linked visitor variables to animal behavioral changes indicative of stress. There are implications for research design: Studies not accounting for visitors' effect on animal welfare risk confounding (visitor) variables distorting their findings. Zoos need methods to measure and minimize effects of visitor behavior and to ensure that there are no hidden variables in research models. This article identifies a previously unreported variable--hourly variation (decrease) in visitor interest--that may impinge on animal welfare and validates a methodology for measuring it. That visitor interest wanes across the course of the day has important implications for animal welfare management; visitor effects on animal welfare are likely to occur, or intensify, during the morning or in earlier visits when visitor interest is greatest. This article discusses this issue and possible solutions to reduce visitor effects on animal well-being. PMID- 17112339 TI - Methods for improving regression analysis for skewed continuous or counted responses. AB - Standard inference procedures for regression analysis make assumptions that are rarely satisfied in practice. Adjustments must be made to insure the validity of statistical inference. These adjustments, known for many years, are used routinely by some health researchers but not by others. We review some of these methods and give an example of their use in a health services study for a continuous and a count outcome. For the continuous outcome, we describe re transformation using the smear factor, accounting for missing cases via multiple imputation and attrition weights and improving results with bootstrap methods. For the count outcome, we describe zero inflated Poisson and negative binomial models and the two-part model to account for overabundance of zero values. Recent advances in computing and software development have produced user-friendly computer programs that enable the data analyst to improve prediction and inference based on regression analysis. PMID- 17112340 TI - Managed behavioral health care carve-outs: past performance and future prospects. AB - As the managed behavioral health care market has matured, behavioral health carve outs have solved many problems facing the delivery of behavioral health services; at the same time, they have exacerbated existing difficulties or created new problems. Carve-outs developed to address rising inpatient behavioral health costs and limited insurance coverage. They are based on the economic principles of economies of specialization, economies of scale, price negotiation, and selection. Literature shows that carve-outs have been successful in lowering costs and maintaining or improving access, but results on their impact on quality of care are mixed. In recent years, carve-outs have evolved to take on new roles within the health system, such as coordinating mental and physical health, addressing fragmented public financing systems, and using market power to implement quality improvement. Although not perfect, carve-outs have been instrumental in addressing long-standing challenges in utilization, access, and cost of behavioral health care. PMID- 17112341 TI - Kinetic analysis of GTP hydrolysis catalysed by the Arf1-GTP-ASAP1 complex. AB - Arf (ADP-ribosylation factor) GAPs (GTPase-activating proteins) are enzymes that catalyse the hydrolysis of GTP bound to the small GTP-binding protein Arf. They have also been proposed to function as Arf effectors and oncogenes. We have set out to characterize the kinetics of the GAP-induced GTP hydrolysis using a truncated form of ASAP1 [Arf GAP with SH3 (Src homology 3) domain, ankyrin repeats and PH (pleckstrin homology) domains 1] as a model. We found that ASAP1 used Arf1-GTP as a substrate with a k(cat) of 57+/-5 s(-1) and a K(m) of 2.2+/ 0.5 microM determined by steady-state kinetics and a kcat of 56+/-7 s(-1) determined by single-turnover kinetics. Tetrafluoroaluminate (AlF4-), which stabilizes complexes of other Ras family members with their cognate GAPs, also stabilized a complex of Arf1-GDP with ASAP1. As anticipated, mutation of Arg-497 to a lysine residue affected kcat to a much greater extent than K(m). Changing Trp-479, Iso-490, Arg-505, Leu-511 or Asp-512 was predicted, based on previous studies, to affect affinity for Arf1-GTP. Instead, these mutations primarily affected the k(cat). Mutants that lacked activity in vitro similarly lacked activity in an in vivo assay of ASAP1 function, the inhibition of dorsal ruffle formation. Our results support the conclusion that the Arf GAP ASAP1 functions in binary complex with Arf1-GTP to induce a transition state towards GTP hydrolysis. The results have led us to speculate that Arf1-GTP-ASAP1 undergoes a significant conformational change when transitioning from the ground to catalytically active state. The ramifications for the putative effector function of ASAP1 are discussed. PMID- 17112342 TI - Mouse Cyp4a isoforms: enzymatic properties, gender- and strain-specific expression, and role in renal 20-hydroxyeicosatetraenoic acid formation. AB - AA (arachidonic acid) hydroxylation to 20-HETE (20-hydroxyeicosatetraenoic acid) influences renal vascular and tubular function. To identify the CYP (cytochrome P450) isoforms catalysing this reaction in the mouse kidney, we analysed the substrate specificity of Cyp4a10, 4a12a, 4a12b and 4a14 and determined sex- and strain-specific expressions. All recombinant enzymes showed high lauric acid hydroxylase activities. Cyp4a12a and Cyp4a12b efficiently hydroxylated AA to 20 HETE with V(max) values of approx. 10 nmol x nmol(-1) x min(-1) and K(m) values of 20-40 microM. 20-Carboxyeicosatetraenoic acid occurred as a secondary metabolite. AA hydroxylase activities were approx. 25-75-fold lower with Cyp4a10 and not detectable with Cyp4a14. Cyp4a12a and Cyp4a12b also efficiently converted EPA (eicosapentaenoic acid) into 19/20-OH- and 17,18-epoxy-EPA. In male mice, renal microsomal AA hydroxylase activities ranged between approx. 100 (NMRI), 45 55 (FVB/N, 129 Sv/J and Balb/c) and 25 pmol x min(-1) x mg(-1) (C57BL/6). The activities correlated with differences in Cyp4a12a protein and mRNA levels. Treatment with 5alpha-dihydrotestosterone induced both 20-HETE production and Cyp4a12a expression more than 4-fold in male C57BL/6 mice. All female mice showed low AA hydroxylase activities (15-25 pmol x min(-1) x mg(-1)) and very low Cyp4a12a mRNA and protein levels, but high Cyp4a10 and Cyp4a14 expression. Renal Cyp4a12b mRNA expression was almost undetectable in both sexes of all strains. Thus Cyp4a12a is the predominant 20-HETE synthase in the mouse kidney. Cyp4a12a expression determines the sex- and strain-specific differences in 20-HETE generation and may explain sex and strain differences in the susceptibility to hypertension and target organ damage. PMID- 17112343 TI - Guidelines for the development of local standards of oral health care for people with dementia. PMID- 17112344 TI - The structure and mode of action of different botulinum toxins. AB - The seven serotypes (A-G) of botulinum neurotoxin (BoNT) are proteins produced by Clostridium botulinum and have multifunctional abilities: (i) they target cholinergic nerve endings via binding to ecto-acceptors (ii) they undergo endocytosis/translocation and (iii) their light chains act intraneuronally to block acetylcholine release. The fundamental process of quantal transmitter release occurs by Ca2+-regulated exocytosis involving sensitive factor attachment protein-25 (SNAP-25), syntaxin and synaptobrevin. Proteolytic cleavage by BoNT-A of nine amino acids from the C-terminal of SNAP-25 disables its function, causing prolonged muscle weakness. This unique combination of activities underlies the effectiveness of BoNT-A haemagglutinin complex in treating human conditions resulting from hyperactivity at peripheral cholinergic nerve endings. In vivo imaging and immunomicroscopy of murine muscles injected with type A toxin revealed that the extended duration of action results from the longevity of its protease, persistence of the cleaved SNAP-25 and a protracted time course for the remodelling of treated nerve-muscle synapses. In addition, an application in pain management has been indicated by the ability of BoNT to inhibit neuropeptide release from nociceptors, thereby blocking central and peripheral pain sensitization processes. The widespread cellular distribution of SNAP-25 and the diversity of the toxin's neuronal acceptors are being exploited for other therapeutic applications. PMID- 17112345 TI - Using translational medicine to understand clinical differences between botulinum toxin formulations. AB - When using botulinum toxin-based products, the physician must decide the optimal location and dose required to alleviate symptoms and improve the patient's quality of life. To deliver effective treatment, the physician needs to understand the importance of accurate target muscle selection and localization and the implications of each product's migration properties when diluted in different volumes. Pre-clinical mouse models of efficacy and safety have been utilized to compare local and distal muscle relaxation effects following defined intramuscular administration. Data from the model allow the products to be ranked based on their propensity for local efficacy versus their distal migration properties. Using standardized dilutions, the non-parallel dose-response curves for the various formulations demonstrate that they have different efficacy profiles. Distal effects were also noted at different treatment doses, which are reflected in the different safety and/or therapeutic margins. Based on these pre clinical data, the safety and therapeutic margin rankings are ordered, largest to smallest, as BOTOX, Dysport and Myobloc. The results of subsequent clinical trials are variable and dose comparisons are inconclusive, thus supporting the regulatory position that the dose units of the individual preparations are unique and cannot be simply converted between products. PMID- 17112346 TI - Clinical value of botulinum toxin in neurological indications. AB - Botulinum toxin type-A (BoNT-A) prevents the release of acetylcholine at cholinergic junctions, thereby causing temporary muscle weakness lasting 3-4 months. It is now widely used to treat a broad range of clinical disorders characterized by muscle hyperactivity. BoNT-A has proved effective in the management of several neurological conditions and, in particular, in the management of movement disorders (e.g. blepharospasm, cervical dystonia, laryngeal dystonia, limb dystonia, hemifacial spasm, focal tics, tremor and other hyperkinetic disorders). As a treatment of spasticity, BoNT-A can improve mobility and dexterity as well as preventing the development of distressing and costly secondary complications. In cerebral palsy, BoNT-A is of value, being able to delay or even avoid surgery until motion patterns have become established. PMID- 17112347 TI - Improvements in healthcare and cost benefits associated with botulinum toxin treatment of spasticity and muscle overactivity. AB - Spasticity is a widespread, disabling form of muscle overactivity affecting patients with central nervous system damage resulting in upper motor neurone syndrome. There is a range of effective therapies for the treatment of spasticity (e.g. physical, anaesthetic, chemodenervation and neurolytic injections, systemic medication and surgery), but all therapies must be based on an individualized, multidisciplinary programme targeted to achieve patient goals. Appropriate therapy should be based on the extent and severity of spasticity, but spasticity and its consequences, regardless of presentation or cause, are commonly treated with systemic agents. This may be ill-advised as systemic treatment is associated with many undesirable effects. In particular, elderly patients with post-stroke spasticity are at risk from the central adverse effects of systemic medication (e.g. sedation and gait disturbance), which make them more susceptible to falling, with an associated increased risk of fracture. The rising costs of fracture care and its sequelae are fast becoming an international problem contributing to high healthcare expenditure. Botulinum toxin type-A (BoNT-A) treatment is highly effective for some of the more common forms of spasticity and muscle overactivity, and has a favourable profile when compared with systemic agents and other focal treatments. Therefore, the clinical benefits of BoNT-A treatment outweigh the apparent high costs of this intervention, showing it to be a cost-effective treatment. PMID- 17112348 TI - Safety and efficacy of botulinum toxin type A following long-term use. AB - Botulinum toxin serotype A (BoNT-A) has long heritage of use leading to confidence in its safety and efficacy. The application of BoNT-A does not lead to persistent histological changes in the nerve terminal or the target muscle. Clinical trials defined the safety and tolerability profile of BoNT-A across common therapeutic indications and showed an incidence of adverse events of approximately 25% in the BoNT-A-treated group compared with 15% in the control group. Focal weakness was the only adverse event to occur more often following BoNT-A treatment. Long-term BoNT-A administration has been assessed in various treatment settings, with the level and duration of BoNT-A efficacy response being maintained over repeated rounds of injection with no major safety concerns. The treatment of children with cerebral palsy often require long-term, repeated, multimuscle BoNT-A injections that lead to the administration of comparably higher toxin doses. Despite the high total body doses used, their distribution over multiple muscles and injection sites means that systemic side effects are rare. Recent formulation changes have reduced the incidence of antibody development following treatment with BOTOX. These findings show long-term BoNT-A treatment to be both safe and efficacious for a wide variety of indications. PMID- 17112349 TI - Landmines, pain, suffering, and the public health: a global challenge. PMID- 17112351 TI - Landmines and landmine injuries: an overview. PMID- 17112352 TI - Acute injury caused by landmines. PMID- 17112353 TI - Neurophysiology of pain from landmine injury. PMID- 17112354 TI - Diagnostic and treatment issues in postamputation pain after landmine injury. PMID- 17112355 TI - Psychological and cultural influences on pain and recovery from landmine injury. PMID- 17112356 TI - Rehabilitation after landmine injury. PMID- 17112358 TI - Pain intensity, emotional state, and personality trait: which comes first? PMID- 17112359 TI - Pain medicine recognized as a specialty in Australia. PMID- 17112360 TI - Pain treatment, drug diversion, and the casualties of war. PMID- 17112361 TI - Postoperative pain significantly influences postoperative blood loss in patients undergoing total knee replacement. AB - BACKGROUND AND OBJECTIVES: Although hypertension has long been recognized as a factor that might increase intraoperative blood losses in major orthopedic surgery, the effects of postoperative pain-induced hypertension on blood losses have not so far been evaluated. The aim of this study was to evaluate the effect of pain on perioperative blood losses of patients undergoing primary total knee replacement (TKR). METHODS: Data from patients participating in a randomized clinical trial comparing intravenous patient-controlled analgesia (PCA) (N = 20) with PCA plus continuous femoral nerve (three-in-one) block (N = 20) or PCA plus continuous posterior lumbar plexus (psoas compartment) block (N = 20) were prospectively and retrospectively collected. Correlations between relevant variables and measured and calculated blood loss, number of transfused unit, and late (96 hours) postoperative hemoglobin were tested by linear regressions. Stepwise regressions for each of the four above-mentioned goals were constructed using a probability to enter of 0.25 and to leave of 0.1. A P < 0.05 was considered significant. RESULTS: At the stepwise regressions there was a significant positive correlation between measured blood losses and morphine consumption from 12 to 18 hours (P = 0.006); between calculated blood loss and preoperative mean arterial blood pressure (P = 0.01) and preoperative hemoglobin value (P = 0.02); and between late postoperative hemoglobin and body weight (P = 0.047). CONCLUSION: In patients undergoing TKR, there is a significant correlation between measured blood loss and morphine consumption from 12 to 18 hours. It is concluded that postoperative pain significantly influences postoperative blood loss in patients undergoing TKR. PMID- 17112362 TI - Affective modulation of pain in substance-dependent veterans. AB - OBJECTIVE: Prior work suggests that positive affect inhibits pain while negative affect facilitates it. The current study sought to determine whether: 1) affective modulation of pain extends to a patient population; 2) cocaine and alcohol dependence influences the pattern of modulation; and 3) affective modulation of pain is mediated by changes in arm temperature. DESIGN: Thirty seven participants with and without substance dependence (14 alcohol, 13 cocaine, 10 none) attended three experimental sessions intended to induce emotions (negative, neutral, positive) by picture-viewing. Following emotion-induction, participants were asked to submerge their arm in 33 degrees F water and keep it there until they reached tolerance. During submersion, pain ratings were made on a mechanical visual analog scale (M-VAS). OUTCOME MEASURES: Latency from submersion to first movement of the M-VAS (pain threshold) and latency to arm removal (pain tolerance) were measured. Arm temperature and manipulation checks for emotion-induction (corrugator electromyogram, heart rate, skin conductance, self-report) were also recorded. RESULTS: Manipulation checks confirmed that targeted affective states were achieved. Pain threshold and tolerance were higher after viewing pleasant pictures than after unpleasant ones. Although arm temperature did vary based on the affect induced, analyses suggested that temperature did not influence pain outcomes. CONCLUSIONS: Affect modulates pain perception in patients and does not appear to be mediated by changes in arm temperature. Additionally, pain modulation was not significantly influenced by cocaine or alcohol dependence. These data are encouraging, because they suggest that nonpharmacological methods of pain modulation may be effective in substance dependent individuals. PMID- 17112363 TI - Impact of a functional restoration program on pain and health-related quality of life in patients with chronic low back pain. AB - OBJECTIVE: Functional restoration programs for chronic low back pain (CLBP) have been shown to be successful in improving function and, to a lesser extent, in reducing pain. The Munich Functional Restoration Program (MFRP) is a 4-week outpatient program designed to reduce pain and to improve health-related quality of life in patients with a long history of CLBP. DESIGN: In a retrospective matched concurrent-controls therapeutic study, 44 patients with CLBP, who had either undergone MFRP or received an outpatient standard treatment (control) after initial evaluation at the pain center, completed questionnaires 1 year after the respective therapy (t1). The following parameters were assessed: health related quality of life with Short Form-36 (SF-36), Pain Disability Index (PDI), Numeric Rating Scale (NRS) for pain, depression with the Center for Epidemiological Studies Depression Test (CES-D), and occupational situation. These data were compared with baseline values assessed by a questionnaire completed before starting the respective treatment (baseline, t0). RESULTS: Compared with control, NRS and PDI were significantly better in patients completing the MFRP. Patients of the MFRP group showed also a significant reduction in CES-D as well as an improvement in three of eight SF-36 subscales. No changes were detected in the control group receiving standard treatment. CONCLUSIONS: Compared with standard treatment, a functional restoration program for CLBP significantly improves some aspects of health-related quality of life. It results in a decrease of pain and pain-related disability even in patients with a long history of CLBP. PMID- 17112364 TI - Chronic pain and the measurement of personality: do states influence traits? AB - STUDY DESIGN: This is a structured evidence-based review of all available studies on the effect of pain, (a state phenomenon) on the measurement of personality characteristics (a trait phenomenon). OBJECTIVES: To determine whether pain treatment changes trait scores. SUMMARY OF BACKGROUND DATA: Recent evidence from the psychiatric literature indicates that the measurement of personality characteristics (traits) can be affected or changed by the presence of state psychiatric disorders, for example, depression. At issue then is whether the measurement of chronic pain patients' (CPPs') trait characteristics is affected by the presence of pain, a state problem. METHODS: Computer and manual literature searches for pain studies that reported a prepain treatment and postpain treatment (test-retest) personality test or inventory score produced 35 such reports. These references were reviewed in detail and information relating to the above problem was abstracted and placed into tabular form. Each report was also categorized as to the type of study it represented according to the guidelines developed by the Agency of Health Care Policy and Research (AHCPR). In addition, a list of 15 quality criteria was utilized to measure the quality of each study. Each study was independently categorized for each criterion as positive (criterion filled), negative (criterion not filled), or not applicable, by two of the authors. Only studies having a quality score of 65% or greater were utilized to formulate the conclusions of this review. The strength and consistency of the evidence represented by the remaining studies were then categorized according to the AHCPR guidelines. Conclusions of this review were based on these results. RESULTS: Of the 35 reports, 32 had quality scores of 65% or greater. According to the AHCPR guidelines, there was a consistent finding that the Minnesota Multiphasic Personality Inventory (MMPI) scores changed (improved) with treatment. In reference to the Millon Behavioral Health Inventory, Locus of Control, the Symptom Checklist-90-Revised (SCL-90-R), trait anxiety, and personality disorders, there were not enough studies to draw conclusions about consistency. In reference to coping/self-efficacy inventories, somatization/illness behavior inventories, and personality questionnaire studies, there was a generally consistent finding that these tests changed (improved) with pain treatment. Overall, of the 32 reports, 92.3% demonstrated a change in trait scores (improvement) with pain treatment. This evidence was categorized as consistent. Finally, 100% of a subgroup of reports (N = 12) that had controlled for pain indicated that there was a relationship between a change in pain scores and a change in trait scores. CONCLUSIONS: Based on the above results, it was concluded that some trait tests and inventories may not be pain state independent. Therefore, caution is warranted in interpreting postpain development personality profiles as being indicative of the true prepain personality structure, if measured by these tests. Why trait scores may change with treatment, confounding test-retest issues, and whether trait tests actually measure what they allegedly measure are discussed. PMID- 17112365 TI - Disparities in pain: ethical issues. PMID- 17112366 TI - Erythromelalgia: a case report and literature review. AB - OBJECTIVE: Erythromelalgia is a rare condition, of uncertain etiology, characterized by episodic erythema, intense burning pain and warmth of the hands and/or feet, and when chronic, associated with significant disability. The diagnosis is based on a thorough history and physical exam during a painful episode along with diagnostic testing to exclude other causes. This paper describes the unique syndrome of erythromelalgia through a case report and literature review. DESIGN: Case presentation and literature review. PATIENTS: A 44-year-old male with erythromelalgia. CONCLUSIONS: Despite multiple treatment options, erythromelalgia is a challenging disease to effectively manage. Early recognition and treatment may offer patients the best probability of achieving remission or significant improvement. PMID- 17112367 TI - Intraperitoneal bupivacaine for postoperative pain relief after laparoscopic cholecystectomy. PMID- 17112368 TI - International society on brain and behaviour: 1st international congress on brain and behaviour hyatt regency hotel, thessaloniki, Greece. 20-23 november, 2003. Abstracts. PMID- 17112369 TI - International Society on Brain and Behaviour: 2nd International Congress on Brain and Behaviour. Thessaloniki, Greece. 17-20 november 2005. Abstracts. PMID- 17112371 TI - Real-time ultrasound-guided catheterisation of the internal jugular vein: a prospective comparison with the landmark technique in critical care patients. AB - INTRODUCTION: Central venous cannulation is crucial in the management of the critical care patient. This study was designed to evaluate whether real-time ultrasound-guided cannulation of the internal jugular vein is superior to the standard landmark method. METHODS: In this randomised study, 450 critical care patients who underwent real-time ultrasound-guided cannulation of the internal jugular vein were prospectively compared with 450 critical care patients in whom the landmark technique was used. Randomisation was performed by means of a computer-generated random-numbers table, and patients were stratified with regard to age, gender, and body mass index. RESULTS: There were no significant differences in gender, age, body mass index, or side of cannulation (left or right) or in the presence of risk factors for difficult venous cannulation such as prior catheterisation, limited sites for access attempts, previous difficulties during catheterisation, previous mechanical complication, known vascular abnormality, untreated coagulopathy, skeletal deformity, and cannulation during cardiac arrest between the two groups of patients. Furthermore, the physicians who performed the procedures had comparable experience in the placement of central venous catheters (p = non-significant). Cannulation of the internal jugular vein was achieved in all patients by using ultrasound and in 425 of the patients (94.4%) by using the landmark technique (p < 0.001). Average access time (skin to vein) and number of attempts were significantly reduced in the ultrasound group of patients compared with the landmark group (p < 0.001). In the landmark group, puncture of the carotid artery occurred in 10.6% of patients, haematoma in 8.4%, haemothorax in 1.7%, pneumothorax in 2.4%, and central venous catheter-associated blood stream infection in 16%, which were all significantly increased compared with the ultrasound group (p < 0.001). CONCLUSION: The present data suggest that ultrasound-guided catheterisation of the internal jugular vein in critical care patients is superior to the landmark technique and therefore should be the method of choice in these patients. PMID- 17112370 TI - Autosomal recessive cerebellar ataxias. AB - Autosomal recessive cerebellar ataxias (ARCA) are a heterogeneous group of rare neurological disorders involving both central and peripheral nervous system, and in some case other systems and organs, and characterized by degeneration or abnormal development of cerebellum and spinal cord, autosomal recessive inheritance and, in most cases, early onset occurring before the age of 20 years. This group encompasses a large number of rare diseases, the most frequent in Caucasian population being Friedreich ataxia (estimated prevalence 2-4/100,000), ataxia-telangiectasia (1-2.5/100,000) and early onset cerebellar ataxia with retained tendon reflexes (1/100,000). Other forms ARCA are much less common. Based on clinicogenetic criteria, five main types ARCA can be distinguished: congenital ataxias (developmental disorder), ataxias associated with metabolic disorders, ataxias with a DNA repair defect, degenerative ataxias, and ataxia associated with other features. These diseases are due to mutations in specific genes, some of which have been identified, such as frataxin in Friedreich ataxia, alpha-tocopherol transfer protein in ataxia with vitamin E deficiency (AVED), aprataxin in ataxia with oculomotor apraxia (AOA1), and senataxin in ataxia with oculomotor apraxia (AOA2). Clinical diagnosis is confirmed by ancillary tests such as neuroimaging (magnetic resonance imaging, scanning), electrophysiological examination, and mutation analysis when the causative gene is identified. Correct clinical and genetic diagnosis is important for appropriate genetic counseling and prognosis and, in some instances, pharmacological treatment. Due to autosomal recessive inheritance, previous familial history of affected individuals is unlikely. For most ARCA there is no specific drug treatment except for coenzyme Q10 deficiency and abetalipoproteinemia. PMID- 17112372 TI - Novel methods for secondary structure determination using low wavelength (VUV) circular dichroism spectroscopic data. AB - BACKGROUND: Circular Dichroism (CD) spectroscopy is a widely used method for studying protein structures in solution. Modern synchrotron radiation CD (SRCD) instruments have considerably higher photon fluxes than do conventional lab-based CD instruments, and hence have the ability to routinely measure CD data to much lower wavelengths. Recently a new reference dataset of SRCD spectra of proteins of known structure, designed to cover secondary structure and fold space, has been produced which includes low wavelength (vacuum ultraviolet - VUV) data. However, the existing algorithms used to calculate protein secondary structures from CD data have not been designed to take optimal advantage of the additional information in these low wavelength data. RESULTS: In this study, we have optimised secondary structure calculation methods based on the low wavelength CD data by examining existing algorithms and secondary structure assignment schemes, and then developing new methods which have produced clear improvements in prediction accuracy, especially for beta-sheet components. We have further shown that if precise measurements of protein concentrations, and therefore spectral magnitudes, are not available, the inclusion of the low wavelength data will significantly improve the analyses. However, we have also demonstrated that the new reference dataset, methods, and assignments can also improve the analyses of conventional circular dichroism data, even if the low wavelength data is not available. CONCLUSION: VUV CD data include important information on protein structure which can be exploited with the algorithms and methodologies described. PMID- 17112373 TI - A convenient scheme for coupling a finite element curvilinear mesh to a finite element voxel mesh: application to the heart. AB - BACKGROUND: In some cases, it may be necessary to combine distinct finite element meshes into a single system. The present work describes a scheme for coupling a finite element mesh, which may have curvilinear elements, to a voxel based finite element mesh. METHODS: The method is described with reference to a sample problem that involves combining a heart, which is defined by a curvilinear mesh, with a voxel based torso mesh. The method involves the creation of a temporary (scaffolding) mesh that couples the outer surface of the heart mesh to a voxel based torso mesh. The inner surface of the scaffolding mesh is the outer heart surface, and the outer surface of the scaffolding mesh is defined by the nodes in the torso mesh that are nearest (but outside of) the heart. The finite element stiffness matrix for the scaffolding mesh is then computed. This stiffness matrix includes extraneous nodes that are then removed, leaving a coupling matrix that couples the original outer heart surface nodes to adjacent nodes in the torso voxel mesh. Finally, a complete system matrix is assembled from the pre-existing heart stiffness matrix, the heart/torso coupling matrix, and the torso stiffness matrix. RESULTS: Realistic body surface electrocardiograms were generated. In a test involving a dipole embedded in a spherical shell, relative error of the scheme rapidly converged to slightly over 4%, although convergence thereafter was relatively slow. CONCLUSION: The described method produces reasonably accurate results and may be best suited for problems where computational speed and convenience have a higher priority than very high levels of accuracy. PMID- 17112374 TI - Determinants for receiving acupuncture for LBP and associated treatments: a prospective cohort study. AB - BACKGROUND: Acupuncture is a frequently used but controversial adjunct to the treatment of chronic low back pain (LBP). Acupuncture is now considered to be effective for chronic LBP and health care systems are pressured to make a decision whether or not acupuncture should be covered. It has been suggested that providing such services might reduce the use of other health care services. Therefore, we explored factors associated with acupuncture treatment for LBP and the relation of acupuncture with other health care services. METHODS: This is a post hoc analysis of a longitudinal prospective cohort study. General practitioners (GPs) recruited consecutive adult patients with LBP. Data on physical function, subjective mood and utilization of health care services was collected at the first consultation and at follow-up telephone interviews for a period of twelve months. RESULTS: A total of 179 (13 %) out of 1,345 patients received acupuncture treatment. The majority of those (59 %) had chronic LBP. Women and elderly patients were more likely to be given acupuncture. Additional determinants of acupuncture therapy were low functional capacity and chronicity of pain. Chronic (vs. acute) back pain OR 1.6 (CL 1.4-2.9) was the only significant disease-related factor associated with the treatment. The strongest predictors for receiving acupuncture were consultation with a GP who offers acupuncture OR 3.5 (CL 2.9-4.1) and consultation with a specialist OR 2.1 (CL 1.9 2.3). After adjustment for patient characteristics, acupuncture remained associated with higher consultation rates and an increased use of other health care services like physiotherapy. CONCLUSION: Receiving acupuncture for LBP depends mostly on the availability of the treatment. It is associated with increased use of other health services even after adjustment for patient characteristics. In our study, we found that receiving acupuncture does not offset the use of other health care resources. A significant proportion of patients who received did not meet the so far only known selection criterion (chonicity). Acupuncture therapy might be a reflection of helplessness in both patients and health care providers. PMID- 17112375 TI - Imaging corticospinal tract connectivity in injured rat spinal cord using manganese-enhanced MRI. AB - BACKGROUND: Manganese-enhanced MRI (MEI) offers a novel neuroimaging modality to trace corticospinal tract (CST) in live animals. This paper expands this capability further and tests the utility of MEI to image axonal fiber connectivity in CST of injured spinal cord (SC). METHODS: A rat was injured at the thoracic T4 level of the SC. The CST was labeled with manganese (Mn) injected intracortically at two weeks post injury. Next day, the injured SC was imaged using MEI and diffusion tensor imaging (DTI) modalities. RESULTS: In vivo MEI data obtained from cervical SC confirmed that CST was successfully labeled with Mn. Ex vivo MEI data obtained from excised SC depicted Mn labeling of the CST in SC sections caudal to the lesion, which meant that Mn was transported through the injury, possibly mediated by viable CST fibers present at the injury site. Examining the ex vivo data from the injury epicenter closely revealed a thin strip of signal enhancement located ventrally between the dorsal horns. This enhancement was presumably associated with the Mn accumulation in these intact fibers projecting caudally as part of the CST. Additional measurements with DTI supported this view. CONCLUSION: Combining these preliminary results collectively demonstrated the feasibility of imaging fiber connectivity in experimentally injured SC using MEI. This approach may play important role in future investigations aimed at understanding the neuroplasticity in experimental SCI research. PMID- 17112377 TI - Evaluation of tandem repeats for MLVA typing of Streptococcus uberis isolated from bovine mastitis. AB - BACKGROUND: Streptococcus uberis is a common cause of bovine mastitis and recommended control measures, based on improved milking practice, teat dipping and antibiotic treatment at drying-off, are poorly efficient against this environmental pathogen. A simple and efficient typing method would be helpful in identifying S.uberis sources, virulent strains and cow to cow transmission. The potential of MLVA (Multiple Loci VNTR Analysis; VNTR, Variable Number of Tandem Repeats) for S. uberis mastitis isolates genotyping was investigated. RESULTS: The genomic sequence of Streptococcus uberis (strain 0104J) was analyzed for potential variable number tandem repeats (VNTRs). Twenty-five tandem repeats were identified and amplified by PCR with DNA samples from 24 S. uberis strains. A set of seven TRs were found to be polymorphic and used for MLVA typing of 88 S. uberis isolates. A total of 82 MLVA types were obtained with 22 types among 26 strains isolated from the milk of mastitic cows belonging to our experimental herd, and 61 types for 62 epidemiologically unrelated strains, i.e. collected in different herds and areas. CONCLUSION: The MLVA method can be applied to S. uberis genotyping and constitutes an interesting complement to existing typing methods. This method, which is easy to perform, low cost and can be used in routine, could facilitate investigations of the epidemiology of S. uberis mastitis in dairy cows. PMID- 17112376 TI - Integration and mining of malaria molecular, functional and pharmacological data: how far are we from a chemogenomic knowledge space? AB - The organization and mining of malaria genomic and post-genomic data is important to significantly increase the knowledge of the biology of its causative agents, and is motivated, on a longer term, by the necessity to predict and characterize new biological targets and new drugs. Biological targets are sought in a biological space designed from the genomic data from Plasmodium falciparum, but using also the millions of genomic data from other species. Drug candidates are sought in a chemical space containing the millions of small molecules stored in public and private chemolibraries. Data management should, therefore, be as reliable and versatile as possible. In this context, five aspects of the organization and mining of malaria genomic and post-genomic data were examined: 1) the comparison of protein sequences including compositionally atypical malaria sequences, 2) the high throughput reconstruction of molecular phylogenies, 3) the representation of biological processes, particularly metabolic pathways, 4) the versatile methods to integrate genomic data, biological representations and functional profiling obtained from X-omic experiments after drug treatments and 5) the determination and prediction of protein structures and their molecular docking with drug candidate structures. Recent progress towards a grid-enabled chemogenomic knowledge space is discussed. PMID- 17112378 TI - Asymptomatic infection with highly pathogenic avian influenza H5N1 in wild birds: how sound is the evidence? AB - BACKGROUND: Widespread deaths of wild birds from which highly pathogenic avian influenza virus H5N1 has been isolated suggest that the virus continues to be lethal to them. However, asymptomatic carriage by some wild birds could allow birds to spread the virus on migration. Confirmation of such carriage is therefore important for the design of mitigation measures for the disease in poultry. DISCUSSION: Two recent papers have reported the isolation of H5N1 from a small number of water birds in China and Russia and have concluded that wild birds can spread the viruses over long distances on migration. However, both papers contain weaknesses in the provision of ornithological and associated data that compromise conclusions that can be reached about the role of wild birds in the spread of H5N1. We describe the weaknesses of these studies and highlight the need for improved methodological description and methodology, where appropriate, and further research. SUMMARY: A rigorous assessment of whether wild birds can carry H5N1 asymptomatically is critical to evaluating the risks of spread by migratory birds on long-distance migration. PMID- 17112379 TI - The CENP-B homolog, Abp1, interacts with the initiation protein Cdc23 (MCM10) and is required for efficient DNA replication in fission yeast. AB - Abp1, and the closely related Cbh1 and Cbh2 are homologous to the human centromere-binding protein CENP-B that has been implicated in the assembly of centromeric heterochromatin. Fission yeast cells lacking Abp1 show an increase in mini-chromosome instability suggesting that Abp1 is important for chromosome segregation and/or DNA synthesis. Here we show that Abp1 interacts with the DNA replication protein Cdc23 (MCM10) in a two-hybrid assay, and that the Deltaabp1 mutant displays a synthetic phenotype with a cdc23 temperature-sensitive mutant. Moreover, genetic interactions were also observed between abp1+ and four additional DNA replication initiation genes cdc18+, cdc21+, orc1+, and orc2+. Interestingly, we find that S phase is delayed in cells deleted for abp1+ when released from a G1 block. However, no delay is observed when cells are released from an early S phase arrest induced by hydroxyurea suggesting that Abp1 functions prior to, or coincident with, the initiation of DNA replication. PMID- 17112380 TI - A critique of the WHO TobReg's "Advisory Note" report entitled: "Waterpipe tobacco smoking: health effects, research needs and recommended actions by regulators". AB - BACKGROUND AND AIM: The World Health Organisation Study Group on Tobacco Product Regulation (TobReg) has issued in 2005 an "Advisory Note" entitled: "Waterpipe Tobacco Smoking: Health Effects, Research Needs and Recommended Actions by Regulators". "Waterpipe" smoking is now considered a global public health threat and the corresponding artefact is actually known in the world under three main terms: hookah, narghile and shisha. This important report, the first ever prepared by WHO on the subject, poses two major problems. On one hand, its bibliographical references dismiss world chief relevant studies. On the other, it contains a certain number of errors of many orders: biomedical, sociological, anthropological and historical. The purpose of the present study is to highlight, one by one, where these weaknesses and errors lie and show how this official report can be considerably improved. RESULTS: We realise that widely advertised early anthropological studies were not taken into consideration whereas they shed a substantial light on this peculiar form of smoking and help understanding its high complexity. As for concrete errors to be found in this report, they deal with the chemistry of smoke, health-related effects, smoking patterns, description and history of the artefact and its use, gender and underage use aspects, prevention and research needs in this field. CONCLUSION: The scientific credibility of an international expert report may be at stake if its recommendations do not rely on sound objective research findings and a comprehensive review of the existing literature. The critical comments in this study will certainly help improve the present WHO report. PMID- 17112381 TI - Volume measures for linkage disequilibrium. AB - BACKGROUND: Defining measures of linkage disequilibrium (LD) that have good small sample properties and are applicable to multiallelic markers poses some challenges. The potential of volume measures in this context has been noted before, but their use has been hampered by computational challenges. RESULTS: We design a sequential importance sampling algorithm to evaluate volume measures on I x J tables. The algorithm is implemented in a C routine as a complement to exhaustive enumeration. We make the C code available as open source. We achieve fast and accurate evaluation of volume measures in two dimensional tables. CONCLUSION: Applying our code to simulated and real datasets reinforces the belief that volume measures are a very useful tool for LD evaluation: they are not inflated in small samples, their definition encompasses multiallelic markers, and they can be computed with appreciable speed. PMID- 17112382 TI - Gene expression profiling following constitutive activation of MEK1 and transformation of rat intestinal epithelial cells. AB - BACKGROUND: Constitutive activation of MEK1 (caMEK) can induce the oncogenic transformation of normal intestinal epithelial cells. To define the genetic changes that occur during this process, we used oligonucleotide microarrays to determine which genes are regulated following the constitutive activation of MEK in normal intestinal epithelial cells. RESULTS: Microarray analysis was performed using Affymetrix GeneChip and total RNA from doxycycline inducible RIEtiCAMEK cells in the presence or absence of doxycycline. MEK-activation induced at least a three-fold difference in 115 gene transcripts (75 transcripts were up regulated, and 40 transcripts were down-regulated). To verify whether these mRNAs are indeed regulated by the constitutive activation of MEK, RT-PCR analysis was performed using the samples from caMEK expressing RIE cells (RIEcCAMEK cells) as well as RIEtiCAMEK cells. The altered expression level of 69 gene transcripts was confirmed. Sixty-one of the differentially expressed genes have previously been implicated in cellular transformation or tumorogenesis. For the remaining 8 genes (or their human homolog), RT-PCR analysis was performed on RNA from human colon cancer cell lines and matched normal and tumor colon cancer tissues from human patients, revealing three novel targets (rat brain serine protease2, AMP deaminase 3, and cartilage link protein 1). CONCLUSION: Following MEK-activation, many tumor-associated genes were found to have significantly altered expression levels. However, we identified three genes that were differentially expressed in caMEK cells and human colorectal cancers, which have not been previously linked to cellular transformation or tumorogenesis. PMID- 17112383 TI - Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism. AB - BACKGROUND: Although understanding of physiological interactions between plasmid DNA and its host is important for vector design and host optimization in many biotechnological applications, to our knowledge, global studies on plasmid-host interactions have not been performed to date even for well-characterized plasmids. RESULTS: Escherichia coli cells, either devoid of plasmid DNA or bearing plasmid pOri1 (with a single ColE1 replication origin) or plasmid pOri2 (with double ColE1 replication origins), were cultured in a chemostat. We used a combination of metabolic flux analysis, DNA microarray and enzyme activity analysis methods to explore differences in the metabolism between these strains. We found that the presence of plasmids significantly influenced various metabolic pathways in the host cells, e.g. glycolysis, the tricarboxylic acid (TCA) cycle and the pentose phosphate (PP) pathway. Expression of rpiA, a gene coding for ribose-5-phosphate isomerase A, was considerably decreased in E. coli carrying a high copy number plasmid relative to E. coli carrying a low copy number plasmid and plasmid-free E. coli. The rpiA gene was cloned into an expression vector to construct plasmid pETrpiA. Following induction of pETrpiA-bearing E. coli, which harbored either pOri1 or pOri2, with isopropyl-beta-D-thiogalactopyranoside (IPTG), the copy number of pOri1 and pOri2 was sigificantly higher than that measured in a host devoid of pETrpiA. CONCLUSION: The presence of plasmids can significantly influence some metabolic pathways in the host cell. We believe that the results of detailed metabolic analysis may be useful in optimizing host strains, vectors and cultivation conditions for various biotechnological purposes. PMID- 17112384 TI - Music notation: a new method for visualizing social interaction in animals and humans. AB - BACKGROUND: Researchers have developed a variety of techniques for the visual presentation of quantitative data. These techniques can help to reveal trends and regularities that would be difficult to see if the data were left in raw form. Such techniques can be of great help in exploratory data analysis, making apparent the organization of data sets, developing new hypotheses, and in selecting effects to be tested by statistical analysis. Researchers studying social interaction in groups of animals and humans, however, have few tools to present their raw data visually, and it can be especially difficult to perceive patterns in these data. In this paper I introduce a new graphical method for the visual display of interaction records in human and animal groups, and I illustrate this method using data taken on chickens forming dominance hierarchies. RESULTS: This new method presents data in a way that can help researchers immediately to see patterns and connections in long, detailed records of interaction. I show a variety of ways in which this new technique can be used: (1) to explore trends in the formation of both group social structures and individual relationships; (2) to compare interaction records across groups of real animals and between real animals and computer-simulated animal interactions; (3) to search for and discover new types of small-scale interaction sequences; and (4) to examine how interaction patterns in larger groups might emerge from those in component subgroups. In addition, I discuss how this method can be modified and extended for visualizing a variety of different kinds of social interaction in both humans and animals. CONCLUSION: This method can help researchers develop new insights into the structure and organization of social interaction. Such insights can make it easier for researchers to explain behavioural processes, to select aspects of data for statistical analysis, to design further studies, and to formulate appropriate mathematical models and computer simulations. PMID- 17112385 TI - Are Canadian General Internal Medicine training program graduates well prepared for their future careers? AB - BACKGROUND: At a time of increased need and demand for general internists in Canada, the attractiveness of generalist careers (including general internal medicine, GIM) has been falling as evidenced by the low number of residents choosing this specialty. One hypothesis for the lack of interest in a generalist career is lack of comfort with the skills needed to practice after training, and the mismatch between the tertiary care, inpatient training environment and "real life". This project was designed to determine perceived effectiveness of training for 10 years of graduates of Canadian GIM programs to assist in the development of curriculum and objectives for general internists that will meet the needs of graduates and ultimately society. METHODS: Mailed survey designed to explore perceived importance of training for and preparation for various aspects of Canadian GIM practice. After extensive piloting of the survey, including a pilot survey of two universities to improve the questionnaire, all graduates of the 16 universities over the previous ten years were surveyed. RESULTS: Gaps (difference between importance and preparation) were demonstrated in many of the CanMEDS 2000/2005 competencies. Medical problems of pregnancy, perioperative care, pain management, chronic care, ambulatory care and community GIM rotations were the medical expert areas with the largest gaps. Exposure to procedural skills was perceived to be lacking. Some procedural skills valued as important for current GIM trainees and performed frequently (example ambulatory ECG interpretation) had low preparation ratings by trainees. Other areas of perceived discrepancy between training and practice included: manager role (set up of an office), health advocate (counseling for prevention, for example smoking cessation), and professional (end of life issues, ethics). CONCLUSION: Graduates of Canadian GIM training programs over the last ten years have identified perceived gaps between training and important areas for practice. They have identified competencies that should be emphasized in Canadian GIM programs. Ongoing review of graduate's perceptions of training programs as it applies to their current practice is important to ensure ongoing appropriateness of training programs. This information will be used to strengthen GIM training programs in Canada. PMID- 17112386 TI - IntNetDB v1.0: an integrated protein-protein interaction network database generated by a probabilistic model. AB - BACKGROUND: Although protein-protein interaction (PPI) networks have been explored by various experimental methods, the maps so built are still limited in coverage and accuracy. To further expand the PPI network and to extract more accurate information from existing maps, studies have been carried out to integrate various types of functional relationship data. A frequently updated database of computationally analyzed potential PPIs to provide biological researchers with rapid and easy access to analyze original data as a biological network is still lacking. RESULTS: By applying a probabilistic model, we integrated 27 heterogeneous genomic, proteomic and functional annotation datasets to predict PPI networks in human. In addition to previously studied data types, we show that phenotypic distances and genetic interactions can also be integrated to predict PPIs. We further built an easy-to-use, updatable integrated PPI database, the Integrated Network Database (IntNetDB) online, to provide automatic prediction and visualization of PPI network among genes of interest. The networks can be visualized in SVG (Scalable Vector Graphics) format for zooming in or out. IntNetDB also provides a tool to extract topologically highly connected network neighborhoods from a specific network for further exploration and research. Using the MCODE (Molecular Complex Detections) algorithm, 190 such neighborhoods were detected among all the predicted interactions. The predicted PPIs can also be mapped to worm, fly and mouse interologs. CONCLUSION: IntNetDB includes 180,010 predicted protein-protein interactions among 9,901 human proteins and represents a useful resource for the research community. Our study has increased prediction coverage by five-fold. IntNetDB also provides easy-to-use network visualization and analysis tools that allow biological researchers unfamiliar with computational biology to access and analyze data over the internet. The web interface of IntNetDB is freely accessible at http://hanlab.genetics.ac.cn/IntNetDB.htm. Visualization requires Mozilla version 1.8 (or higher) or Internet Explorer with installation of SVGviewer. PMID- 17112387 TI - Inflammation in sputum relates to progression of disease in subjects with COPD: a prospective descriptive study. AB - BACKGROUND: Inflammation is considered to be of primary pathogenic importance in COPD but the evidence on which current understanding is based does not distinguish between cause and effect, and no single mechanism can account for the complex pathology. We performed a prospective longitudinal study of subjects with COPD that related markers of sputum inflammation at baseline to subsequent disease progression. METHODS: A cohort of 56 patients with chronic bronchitis was characterized in the stable state at baseline and after an interval of four years, using physiological measures and CT densitometry. Sputum markers of airway inflammation were quantified at baseline from spontaneously produced sputum in a sub-group (n = 38), and inflammation severity was related to subsequent disease progression. RESULTS: Physiological and CT measures indicated disease progression in the whole group. In the sub-group, sputum myeloperoxidase correlated with decline in FEV1 (rs = -0.344, p = 0.019, n = 37). LTB4 and albumin leakage correlated with TLCO decline (rs = -0.310, p = 0.033, rs = -0.401, p = 0.008, respectively, n = 35) and IL-8 correlated with progression of lung densitometric indices (rs = -0.464, p = 0.005, n = 38). CONCLUSION: The data support a principal causative role for neutrophilic inflammation in the pathogenesis of COPD and suggest that the measurement of sputum inflammatory markers may have a predictive role in clinical practice. PMID- 17112389 TI - Expressive writing for high-risk drug dependent patients in a primary care clinic: a pilot study. AB - BACKGROUND: Previous research has shown that expressive writing is beneficial in terms of both physical and emotional health outcomes. This study aimed to investigate the effectiveness and acceptability of a brief expressive writing intervention for high-risk drug dependent patients in a primary care clinic, and to determine the relationship between linguistic features of writing and health outcomes. METHODS: Participants completed four 15-minute expressive writing tasks over a week, in which they described their thoughts and feelings about a recent stressful event. Self-report measures of physical (SF-12) and psychological health (DASS-21) were administered at baseline and at a two-week follow-up. Fifty three participants were recruited and 14 (26%) completed all measures. RESULTS: No statistically significant benefits in physical or psychological health were found, although all outcomes changed in the direction of improvement. The intervention was well-received and was rated as beneficial by participants. The use of more positive emotion words in writing was associated with improvements in depression and stress, and flexibility in first person pronoun use was associated with improvements in anxiety. Increasing use of cognitive process words was associated with worsening depressive mood. CONCLUSION: Although no significant benefits in physical and psychological health were found, improvements in psychological wellbeing were associated with certain writing styles and expressive writing was deemed acceptable by high-risk drug dependent patients. Given the difficulties in implementing psychosocial interventions in this population, further research using a larger sample is warranted. PMID- 17112388 TI - Ultrastructural analysis of development of myocardium in calreticulin-deficient mice. AB - BACKGROUND: Calreticulin is a Ca2+ binding chaperone of the endoplasmic reticulum which influences gene expression and cell adhesion. The levels of both vinculin and N-cadherin are induced by calreticulin expression, which play important roles in cell adhesiveness. Cardiac development is strictly dependent upon the ability of cells to adhere to their substratum and to communicate with their neighbours. RESULTS: We show here that the levels of N-cadherin are downregulated in calreticulin-deficient mouse embryonic hearts, which may lead to the disarray and wavy appearance of myofibrils in these mice, which we detected at all investigated stages of cardiac development. Calreticulin wild type mice exhibited straight, thick and abundant myofibrils, which were in stark contrast to the thin, less numerous, disorganized myofibrils of the calreticulin-deficient hearts. Interestingly, these major differences were only detected in the developing ventricles while the atria of both calreticulin phenotypes were similar in appearance at all developmental stages. Glycogen also accumulated in the ventricles of calreticulin-deficient mice, indicating an abnormality in cardiomyocyte metabolism. CONCLUSION: Calreticulin is temporarily expressed during heart development where it is required for proper myofibrillogenesis. We postulate that calreticulin be considered as a novel cardiac fetal gene. PMID- 17112390 TI - Concurrent development of testicular seminoma and choriocarcinoma of the superior mediastinum, presented as cervical mass: a case report and implications about pathogenesis of germ-cell tumours. AB - BACKGROUND: Synchronous presentation of more than one germ cell tumours of different histology in the same patient is considered to be very rare. In these cases of multiple germ cell tumours, strong theoretical and clinical data suggest an underlying common pathogenetic mechanism concerning genetic instability or abnormalities during the pluripotent embryonic differentiation and maturation of the germ cell. CASE PRESENTATION: A 25 year-old young man presented with an enlarging, slightly painful left cervical mass. Despite the initial disorientation of the diagnosis to a possible thyroid disorder, the patient underwent complete surgical resection of the mass revealing mediastinal choriocarcinoma. Subsequent ultrasound of the scrotum indicated the presence of a small lobular node in the upper pole of the left testicle and the patient underwent radical left inguinal orchiectomy disclosing a typical seminoma. Based on these results, the patient received 4 cycles of Bleomycin, Etoposide and Platinum chemotherapy experiencing only mild toxicity and resulting in complete ongoing clinical and biochemical remission. CONCLUSION: The pathogenesis of concurrent germ cell tumours in the same patient remains an area of controversy. Although the genetic instability of the pluripotent germ cell offers an adequate explanation, the possibility of metastasis from the primary, less differentiated tumour to a distant location as a more mature subtype cannot be excluded. Possible development of a metastatic site of different histology and thus biological behaviour (e.g choriocarcinoma) should be anticipated. Furthermore, urologists, pathologists and medical oncologists should be meticulous in the original pathological diagnosis in these patients, since there is a significant frequency of germ cell tumours with mixed or overlapping histological elements with diverse potential of evolution and differentiation. PMID- 17112391 TI - Adiponectin, type 2 diabetes and the metabolic syndrome: lessons from human genetic studies. AB - Adiponectin, a protein exclusively secreted by adipose tissue but present at low levels in obesity, is now widely recognised as a key determinant of insulin sensitivity and of protection against obesity-associated metabolic syndrome. In this review we explain how genetic findings have contributed to a better understanding of the physiological role of adiponectin in humans. The adiponectin encoding gene, ADIPOQ (ACDC), is very polymorphic: many frequent exonic synonymous, intronic and promoter single-nucleotide polymorphisms (SNPs) have been identified, as well as a few rare exonic amino acid substitutions. Several of these variations additively contribute to the modulation of adiponectin level and function, and associate with insulin sensitivity, type 2 diabetes and vascular complications of obesity. PMID- 17112392 TI - Parotid abscess: a five-year review--clinical presentation, diagnosis and management. AB - Parotid abscess is an uncommon complication of suppurative infection of the parotid gland parenchyma, commonly bacterial or viral. Ductal ectasis, primary parenchymal involvement, or infection of the intraparotid or periparotid lymph nodes can result in abscess formation. Parotid abscess may arise from ductal ectasis, primary parenchymal involvement, or infection of the subcapsular lymph nodes. The operative records for all the patients who underwent surgeries in the Department of Otorhinolaryngology, Head and Neck Surgery of the National University Hospital, Kuala Lumpur, Malaysia between January 2001 and December 2005 were retrospectively reviewed. Our case series comprises 15 patients, with 10 males and five females with a median age at presentation of 51 years old. Diabetes mellitus is a significant comorbid factor, with six patients being diabetics. Among the diabetics, two patients presented with facial nerve palsy and one of them also died due to overwhelming septicaemia. Here, we discuss the presenting symptoms, predisposing factors, investigations, microbiology and complications of this condition. PMID- 17112393 TI - Role of stapes surgery in improving hearing loss caused by otosclerosis. AB - The aim of our study was to evaluate the functional results of stapes surgery and to compare the effectiveness of small fenestra stapedotomy with that of total stapedectomy in improving hearing in patients affected by otosclerosis. Three hundred and fifty-seven consecutive ears, in 265 patients affected by otosclerosis, underwent surgery. All cases underwent either primary small fenestra stapedotomy (group A, 196/357, 54.91 per cent) or stapedectomy (group B, 161/357, 45.09 per cent). After surgery, 256/357 (71.71 per cent) cases showed a 0-20 dB gap. There were no significant differences in hearing results between the two groups at either early or late post-operative assessment. The mean post operative pure tone average and air-bone gap results were slightly greater for group B than for group A, at both early and late post-operative assessments, but these differences were not statistically significant. Therefore, in group A, the mean pure tone average at 4 kHz significantly improved, from 56.60 to 47.66 dB at early post-operative assessment and to 52.98 dB at late post-operative assessment. Our study suggests that the technique of microtomy of the oval window is able to improve hearing results especially at high frequencies. PMID- 17112394 TI - Myasthenia gravis mimicking unilateral vocal fold paralysis at presentation. AB - OBJECTIVES: To demonstrate the importance of detailed clinical analysis in the differential diagnosis of unilateral vocal fold paralysis, and to provide an update on current knowledge and treatment of myasthenia gravis. CASE REPORT: A female patient presented with left unilateral vocal fold immobility. Diagnostic investigation revealed a 10 mm thyroid adenoma, but no other abnormality likely to cause unilateral vocal fold paralysis. Follow-up flexible endoscopy at three months showed laryngeal remobilisation with persistent left vocal fold bowing and vertical asymmetry of the vocal folds on phonation. Over the following months, voice quality varied between normal and breathy, with the breathy periods lasting from three days to one month. Laryngeal electromyography (EMG) showed a slight bilateral paradoxical activation of both posterior crico-arytenoid muscles on phonation. Magnetic resonance imaging of the brain and brainstem was normal. A diagnostic test for myasthenia gravis with intravenous edrophonium bromide (Tensilon) lead to an immediate improvement in voice quality. The patient was subsequently treated with pyridostigmine bromide, with complete resolution of dysphonia. CONCLUSIONS: Myasthenia gravis affecting the larynx may mimic unilateral vocal fold paresis or paralysis. A personal or family history of auto immune disease, fluctuating symptoms, motor deficits in cranial nerve territories, and normal or subnormal laryngeal EMG results should lead the physician to reconsider a diagnosis of idiopathic unilateral vocal fold paralysis and to perform specific testing. PMID- 17112395 TI - An alternative strategy for universal infant hearing screening in tertiary hospitals with a high delivery rate, within a developing country, using transient evoked oto-acoustic emissions and brainstem evoked response audiometry. AB - OBJECTIVE: To formulate an alternative strategy for universal infants hearing screening in an Indian tertiary referral hospital with a high delivery rate, which could be extended to similar situations in other developing countries. The system should be able to diagnose, in a timely fashion, all infants with severe and profound hearing losses. METHODS: One thousand newborn were randomly selected. All underwent testing with transient evoked oto-acoustic emissions (TEOAE) in the first 48 hours of life. All TEOAE failures were followed up and repeat tests were performed at three weeks, three months and six months of age. Infants with acceptable TEOAE results at any of the four ages were discharged from the study. Infants with unacceptable TEOAE results at all the four ages underwent brainstem evoked response audiometry and oto-endoscopy. The 'pass rate' for TEOAE testing was calculated for all four ages. The time taken to perform TEOAE and brainstem evoked response audiometry was recorded for all subjects. These recordings were statistically analysed to find the most suitable strategy for universal hearing screening in our hospital. RESULTS: The pass rate for TEOAE was 79.0 per cent at < or =48 hours, 85.0 per cent at three weeks, 97.0 per cent at three months and 98.0 per cent at six months. The average time taken to perform the test was 12 minutes for TEOAE and 27 minutes for brainstem evoked response audiometry. Obstructed and collapsed external auditory canals were the two factors that significantly affected the specificity of TEOAE in infants < or =48 hours old. CONCLUSION: The concept of screening all neonates within the first 48 hours of life is impractical because the specificity of TEOAE is lowest at that age. Many false positive results are generated, such that a larger number must undergo brainstem evoked response audiometry, wasting time and resources. This can easily be avoided by delaying TEOAE screening until three months of age, when it has a substantially lower false positive outcome. We expect that implementation of this alternative strategy in our hospital will maximise the benefits of such a programme. PMID- 17112396 TI - Reduction meatoplasty with a post-auricular island flap. AB - The importance of an adequate meatoplasty is often emphasised in mastoid surgery. However, bigger is not always better, as an excessively widened external meatus can be cosmetically unacceptable, provide little extra benefit for cleaning, be an obstacle to the good fitting of hearing aids and expose the mastoid cavity to exaggerated caloric effect. The problems created by an overly large meatus can occasionally be difficult to manage, prompting consideration of reduction of the meatus. We describe the use of a pedicled, post-auricular skin flap to achieve reduction of an excessively large meatus. PMID- 17112397 TI - Use of a rectal snare to remove a hypopharyngeal haemangioma. AB - We describe in this case report a new technique for treatment of hypopharyngeal haemangioma, using the surgical diathermy snare. The snare was easily introduced through the direct laryngoscope, without any difficulties. The procedure was simple, rapid and involved minimal bleeding. We also discuss the histological types of haemangioma, clinical picture, radiological findings and other modalities of treatment. PMID- 17112398 TI - A peripheral nerve sheath tumour as a cause of nasal obstruction. AB - Neurogenic tumours form a very small percentage of all neoplastic lesions of the head and neck region. However, the head and neck region is by far the most common location for benign peripheral nerve tumours. Several cases involving the nose have been sporadically documented throughout the medical literature. We present a rare case of a solitary neurofibroma arising from the lateral nasal wall of a 68 year-old woman. En bloc surgical resection of the mass was achieved by the lateral rhinotomy approach. The clinical significance of this case report is due to its rare site. In recent medical literature, there has been only one report concerning a solitary neurofibroma arising from the inferior turbinate. This case also highlights the importance of considering this clinical entity in the differential diagnosis when encountering a unilateral soft tissue mass in the nasal cavity. PMID- 17112399 TI - Comparative study of palatine tonsil histology in mammals, with special reference to tonsillar salivary glands. AB - The authors studied the histology of palatine tonsils from various mammals. Special reference was made to tonsillar salivary glands, which are usually rudimentary in humans. Tonsillar salivary glands were a prominent feature in all categories of mammals studied, apart from humans. As far as the authors were aware, no previous study had focussed on tonsillar salivary glands in mammals. The putative physiological implications are discussed. PMID- 17112400 TI - Internet-based cognitive behaviour therapy for symptoms of depression and anxiety: a meta-analysis. AB - BACKGROUND: We studied to what extent internet-based cognitive behaviour therapy (CBT) programs for symptoms of depression and anxiety are effective. METHOD: A meta-analysis of 12 randomized controlled trials. RESULTS: The effects of internet-based CBT were compared to control conditions in 13 contrast groups with a total number of 2334 participants. A meta-analysis on treatment contrasts resulted in a moderate to large mean effect size [fixed effects analysis (FEA) d=0.40, mixed effects analysis (MEA) d=0.60] and significant heterogeneity. Therefore, two sets of post hoc subgroup analyses were carried out. Analyses on the type of symptoms revealed that interventions for symptoms of depression had a small mean effect size (FEA d=0.27, MEA d=0.32) and significant heterogeneity. Further analyses showed that one study could be regarded as an outlier. Analyses without this study showed a small mean effect size and moderate, non-significant heterogeneity. Interventions for anxiety had a large mean effect size (FEA and MEA d=0.96) and very low heterogeneity. When examining the second set of subgroups, based on therapist assistance, no significant heterogeneity was found. Interventions with therapist support (n=5) had a large mean effect size, while interventions without therapist support (n=6) had a small mean effect size (FEA d=0.24, MEA d=0.26). CONCLUSIONS: In general, effect sizes of internet-based interventions for symptoms of anxiety were larger than effect sizes for depressive symptoms; however, this might be explained by differences in the amount of therapist support. PMID- 17112401 TI - Adjuvant occupational therapy for work-related major depression works: randomized trial including economic evaluation. AB - BACKGROUND: Major depression has far-reaching consequences for work functioning and absenteeism. In most cases depression is treated by medication and clinical management. The addition of occupational therapy (OT) might improve outcome. We determined the cost-effectiveness of the addition of OT to treatment as usual (TAU). METHOD: Sixty-two adults with major depression and a mean absenteeism of 242 days were randomized to TAU (out-patient psychiatric treatment) or TAU+OT [6 months, including (i) diagnostic phase with occupational history and work reintegration plan, and (ii) therapeutic phase with individual sessions and group sessions]. Main outcome domains were depression, work resumption, work stress and costs. Assessments were at baseline and at 3, 6, 12 and 42 months. RESULTS: The addition of OT to TAU: (i) did not improve depression outcome, (ii) resulted in a reduction in work-loss days during the first 18 months, (iii) did not increase work stress, and (iv) had a 75.5% probability of being more cost-effective than TAU alone. CONCLUSION: Addition of OT to good clinical practice does not improve depression outcome, improves productivity without increasing work stress and is superior to TAU in terms of cost-effectiveness. PMID- 17112402 TI - Assessment of complex mental activity across the lifespan: development of the Lifetime of Experiences Questionnaire (LEQ). AB - BACKGROUND: Brain reserve is a property of the central nervous system related to complex mental activity which may mediate the course and clinical expression of brain injury. Since there is no instrument that comprehensively assesses complex mental activity through the lifespan, we developed and tested the Lifetime of Experiences Questionnaire (LEQ) in a prospective study of healthy ageing. METHOD: The LEQ assesses educational, occupational and cognitive lifestyle activities at different stages through life. Test-retest, item analysis and Item Response Theory (IRT) were used to determine reliability. Dimensionality was evaluated using factor analysis. Validity was established through IRT analysis of test performance, correlation with an extant contemporaneous instrument (Cognitive Activities Scale; CAS) and prediction of global cognitive change over 18 months controlling for age, baseline cognition and hypertension. RESULTS: In a sample of healthy older individuals (n=79) the LEQ was found to be consistent, coherent and discriminate between individuals with high and low mental activity levels. Factor analysis revealed a dominant factor which loaded heavily on education, occupation and leisure activity. Total LEQ was significantly correlated with the CAS. Furthermore, individuals with higher LEQ scores showed less cognitive decline over 18 months, independent of covariates (r=0.37, p=0.003). CONCLUSIONS: The LEQ is a reliable and valid instrument for assessing complex lifespan mental activity which is protective against cognitive decline. The LEQ is therefore proposed as a useful tool for estimating brain reserve in older individuals and further development is anticipated. PMID- 17112403 TI - Quality of life as an outcome indicator in patients with seasonal affective disorder: results from the Can-SAD study. AB - BACKGROUND: Although a host of studies have now examined the relationship between quality of life (QoL) and non-seasonal depression, few have measured QoL in seasonal affective disorder (SAD). We report here on results from the Can-SAD trial, which assessed the impact of treatment with either antidepressant medication or light therapy upon QoL in patients diagnosed with SAD. METHOD: This Canadian double-blind, multicentre, randomized controlled trial included 96 patients who met strict diagnostic criteria for SAD. Eligible patients were randomized to 8 weeks of treatment with either: (1) 10000 lux light treatment and a placebo capsule or (2) 100 lux light treatment (placebo light) and 20 mg fluoxetine. QoL was measured with the Quality of Life Enjoyment and Satisfaction Questionnaire (Q-LES-Q) and the Medical Outcomes Study (MOS) Short-Form General Health Survey (SF-20) at baseline and 8 weeks. RESULTS: Both intervention groups showed significant improvement in QoL over time with no significant differences being detected by treatment condition. Q-LES-Q scores increased significantly in seven of eight domains, with the average scores rising from 48 x 0 (S.D.=10 x 7) at baseline to 69 x 1 (S.D.=15 x 6) at week 8. Treatment-related improvement in QoL was strongly associated with improvement in depression symptoms. DISCUSSION: Patients with SAD report markedly impaired QoL during the winter months. Treatment with light therapy or antidepressant medication is associated with equivalent marked improvement in perceived QoL. Studies of treatment interventions for SAD should routinely include broader indices of patient outcome, such as the assessment of psychosocial functioning or life quality. PMID- 17112404 TI - Sympathetically evoked Ca2+ signaling in arterial smooth muscle. AB - The sympathetic nervous system plays an essential role in the control of total peripheral vascular resistance and blood flow, by controlling the contraction of small arteries. Perivascular sympathetic nerves release ATP, norepinephrine (NE) and neuropeptide Y. This review summarizes our knowledge of the intracellular Ca2+ signals that are activated by ATP and NE, acting respectively on P2X1 and alpha1-adrenoceptors in arterial smooth muscle. Each neurotransmitter produces a unique type of post-synaptic Ca2+ signal and associated contraction. The neural release of ATP and NE is thought to vary markedly with the pattern of nerve activity, probably reflecting both pre- and post-synaptic mechanisms. Finally, we show that Ca2+ signaling during neurogenic contractions activated by trains of sympathetic nerve fiber action potentials are in fact significantly different from that elicited by simple bath application of exogenous neurotransmitters to isolated arteries (a common experimental technique), and end by identifying important questions remaining in our understanding of sympathetic neurotransmission and the physiological regulation of contraction of small arteries. PMID- 17112405 TI - Cysteinyl leukotriene receptor 1 is involved in N-methyl-D-aspartate-mediated neuronal injury in mice. AB - AIM: To determine whether cysteinyl leukotriene receptor 1 (CysLT1 receptor) is involved in N-methyl-D-aspartate (NMDA)-induced excitotoxic injury in the mouse brain. METHODS: Brain injury was induced by NMDA microinjection (50-150 nmol in 0.5 microL) into the cerebral cortex. The changes in CysLT1 receptor expression 24 h after NMDA injection and the effects of a CysLT1 receptor antagonist, pranlukast (0.01 and 0.1 mg/kg), an NMDA receptor antagonist, ketamine (30 mg/kg), and an antioxidant, edaravone (9 mg/kg) were observed. RESULTS: In the NMDA-injured brain, the CysLT1 receptor mRNA, and protein expression were upregulated, and the receptor was mainly localized in the neurons and not in the astrocytes. Pranlukast, ketamine and edaravone decreased NMDA-induced injury; pranlukast (0.1 mg/kg) and ketamine inhibited the upregulated expression of the CysLT1 receptor. CONCLUSION: CysLT1 receptor expression in neurons is upregulated after NMDA injection, and NMDA-induced responses are inhibited by CysLT1 receptor antagonists, indicating that the increased CysLT1 receptor is involved in NMDA excitotoxicity. PMID- 17112406 TI - Kinetic changes and modulation by carbamazepine on voltage-gated sodium channels in rat CA1 neurons after epilepsy. AB - AIM: To study whether the functional properties of sodium channels, and subsequently the channel modulation by carbamazepine (CBZ) in hippocampal CA1 neurons can be changed after epileptic seizures. METHODS: We used the acutely dissociated hippocampal CA1 pyramidal cells from epilepsy model rats 3 weeks and 3 months respectively after kainate injection, and whole-cell voltage-clamp techniques. RESULTS: After long-term epileptic seizures, both sodium channel voltage-dependence of activation and steady-state inactivation shifted to more hyperpolarizing potentials, which resulted in the enlarged window current; the membrane density of sodium current decreased and the time constant of recovery from inactivation increased. CBZ displayed unchanged efficacy on sodium channels, with a similar binding rate to them, except that at higher concentrations, the voltage shift of inactivation was reduced. For the short-term kainate model rats, no differences were detected between the control and epilepsy groups. CONCLUSION: These results indicate that the properties of sodium channels in acutely dissociated hippocampal neurons could be changed following long-term epilepsy, but the alternation might not be enough to induce the channel resistance to CBZ. PMID- 17112407 TI - Intrathecal administration of Cav3.2 and Cav3.3 antisense oligonucleotide reverses tactile allodynia and thermal hyperalgesia in rats following chronic compression of dorsal root of ganglion. AB - AIM: The present study aimed to elucidate the role of T-subtype calcium channels (Cav3.1, Cav3.2, and Cav3.3) in the pathogenesis of neuropathic pain at spinal level. METHODS: The chronic compression of the dorsal root ganglion (CCD) rat model was adopted. The antisense oligonucleotide of Cav3.1, Cav3.2, and Cav3.3 or normal saline (NS) were intrathecally administered twice per day from the first day to the fourth day after operation. Paw mechanical withdrawal threshold and paw thermal withdrawal latency were measured to evaluate the tactile allodynia and thermal hyperalgesia, respectively. RESULTS: CCD rats developed reliable tactile allodynia and thermal hyperalgesia after operation. Intrathecal administration of antisense oligonucleotide of Cav3.2 and Cav3.3 significantly relieved tactile allodynia and thermal hyperalgesia in CCD rats, but not Cav3.1. CONCLUSION: Cav3.2 and Cav3.3 subtype calcium channels in the spinal cord may play an important role in the pathogenesis of neuropathic pain, which may contribute to the management of the neuropathic pain. PMID- 17112408 TI - Cysteinyl leukotriene receptors CysLT1 and CysLT2 are upregulated in acute neuronal injury after focal cerebral ischemia in mice. AB - AIM: To determine whether cysteinyl leukotriene receptors (CysLT1 and CysLT2) are upregulated in acute neuronal injury after focal cerebral ischemia in mice, and to confirm CysLT1 receptor localization. METHODS: After permanent focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO), neurological deficits and neuron loss were determined at various time points within 48 h. The mRNA expressions of CysLT1 and CysLT2 receptors in the brain were analyzed by RT PCR. CysLT1 receptor localization was detected by double immunofluorescence. RESULTS: Neurological deficits and neuron loss were found 6, 24 and 48 h after MCAO. The mRNA expressions of both CysLT1 and CysLT2 receptors were upregulated in the ischemic hemisphere 1, 24, and 48 h after MCAO with peaks at 24 h. The CysLT1 receptor was selectively localized in neurons 24 h after MCAO. CONCLUSION: CysLT1 and CysLT2 receptors are upregulated in acute neuronal injury after focal cerebral ischemia, and the CysLT1 receptor is localized in neurons after ischemia. PMID- 17112409 TI - Interaction between hydrogen sulfide/cystathionine gamma-lyase and carbon monoxide/heme oxygenase pathways in aortic smooth muscle cells. AB - AIM: To investigate the interaction between hydrogen sulfide (H2S)/cystathionine gamma-lyase (CSE) and carbon monoxide (CO)/heme oxygenase (HO) pathways in aortic smooth muscle cells (ASMC). METHODS: The ASMCs were divided into the following groups: (1) the control group; (2) the zinc protoporphyrin (ZnPP) 20 micromol/L group; (3) the propargylglycine (PPG) 2 mmol/L, 4 mmol/L and 10 mmol/L groups; and (4) the sodium hydrosulfide (NaHS) 1 x 10(-5) mol/L, 1 x 10(-4) mol/L and 1 x 10(-3) mol/L groups. Each of the groups was further divided into 6 h, 12 h, 18 h and 24 h subgroups. The CO level, represented by carboxyhemoglobin (HbCO) content was measured using a spectrophotometric method and H2S content was detected by a sensitive electrode method. CSE and HO-1 expressions were detected by Western blotting. RESULTS: The H2S content in the medium and CSE expression by ASMC were markedly increased by ZnPP compared with the control group. HbCO content in the medium and HO-1 expression by the ASMC started strengthening following 24 h treatment with PPG at 2 mmol/L, but were further strengthened following 18 h and 24 h treatment with PPG at 4 mmol/L compared with the controls (P < 0.01). PPG at 10 mmol/L increased the HbCO level in the medium following 18 h treatment and increased HO-1 expression by the ASMC following 12 h treatment. Moreover, NaHS at 1 x 10(-5) mol/L and 1 x 10(-4) mol/L decreased the HbCO level in the medium and HO-1 expression by the ASMC after 6 h and 12 h treatment, while NaHS at 1 x 10( 3) mol/L decreased them at all time points of the treatments. CONCLUSION: The results suggested that endogenous CO/HO and H2S/CSE pathways inhibited each other in ASMC under physiological conditions. PMID- 17112410 TI - Caveolin-1 is important for nitric oxide-mediated angiogenesis in fibrin gels with human umbilical vein endothelial cells. AB - AIM: The role of caveolin-1 (Cav-1) in angiogenesis remains poorly understood. The endothelial nitric oxide (NO) synthase (eNOS), a caveolin-interacting protein, was demonstrated to play a predominant role in vascular endothelial growth factor (VEGF) -induced angiogenesis. The purpose of our study was to examine the role of Cav-1 and the eNOS complex in NO-mediated angiogenesis. METHODS: Human umbilical vein endothelial cells (HUVEC) were isolated and cultured in 3-D fibrin gels to form capillary-like tubules by VEGF stimulation. The expression of Cav-1 and eNOS was detected by semiquantitative RT-PCR. The HUVEC were treated with antisense oligonucleotides to downregulate Cav-1 expression. Both transduced and non-infected HUVEC were cultured in fibrin gels in the presence or absence of VEGF (20 ng/mL) and NG-nitro-L-arginine methyl ester (L-NAME; 5 mmol/L). NO was measured using a NO assay kit and capillary-like tubules were quantified by tubule formation index using the Image J program. RESULTS: RT-PCR analysis revealed that Cav-1 levels steadily increased in a time dependent manner and reached their maximum after 5 d of incubation, but there were no obvious changes in eNOS mRNA expression in response to VEGF in the fibrin gel model. VEGF (20 ng/mL) can promote NO production and the formation of capillary-like tubules, and this promoting effect of VEGF was blocked by the addition of L-NAME (5 mmol/L). When transduced HUVEC with the antisense Cav-1 oligonucleotides were plated in the fibrin gels, the capillary-like tubules were significantly fewer than those of the non-infected cells. The capillary-like tubules formation and NO production of transduced HUVEC with the antisense Cav-1 oligonucleotides cultured in fibrin gels showed no responses to the addition of VEGF (20 ng/mL) and L-NAME (5.0 mmol/L). CONCLUSION: NO was a critical angiogenic mediator in this model. Cav-1 was essential for NO-mediated angiogenesis and may be an important target of anti-angiogenesis therapy. PMID- 17112411 TI - Synergism of hydrochlorothiazide and nitrendipine on reduction of blood pressure and blood pressure variability in spontaneously hypertensive rats. AB - AIM: To investigate the possible synergism of hydrochlorothiazide and nitrendipine on reducing both blood pressure (BP) and blood pressure variability (BPV) in spontaneously hypertensive rats (SHR). METHODS: Seventy animals were randomly divided into seven groups. The doses were 5 and 10 mg/kg for nitrendipine, 10 and 20 mg/kg for hydrochlorothiazide and 10 + 5, 20 + 10 mg/kg, respectively, for the combination of these two drugs and 0.8% carboxymethylcellulose as control. The drugs were given via a catheter of gastric fistula. BP was then continuously recorded for 5 h from 1 h before drug administration to the end of 4th hour after drug administration, in conscious and freely moving rats. RESULTS: The effects on both BP and BPV reduction of the combination of hydrochlorothiazide and nitrendipine were greater than the single drug in SHR. The two drugs possessed an obvious synergism on both systolic blood pressure (q = 1.79 with small dose and q = 1.23 with large dose) and systolic blood pressure variability reduction (q = 1.79 with small dose and q = 1.39 with large dose) in SHR. CONCLUSION: The present work clearly demonstrated that there was a synergistic effect between hydrochlorothiazide and nitrendipine in lowering and stabilizing BP in SHR. PMID- 17112412 TI - Phorbol-induced surface expression of NR2A subunit homologues in HEK293 cells. AB - AIM: N-methyl-D-aspartate receptors (NMDAR) are heteromeric complexes primarily assembled from NR1 and NR2 subunits. In normal conditions, NR2 subunits assemble into homodimers in the endoplasmic reticulum (ER). These homodimers remain in the ER until they coassemble with NR1 dimers and are trafficked to the cell surface. However, it still remains unclear whether functional homomeric NMDAR exist in physiological or pathological conditions. METHODS: We transfected GFP-NR2A alone into HEK293 cells, treated the cells with PKC activator 12-myristate-13 acetate (PMA), and then detected surface NR2A subunits with a live cell immunostaining method. We also used a series of NR2A mutants with a partial deletion of its C terminus to identify the regions that are involved in the PMA-mediated surface expression of NR2A subunits. RESULTS: NR2A subunits were expressed on the cell membrane after incubation with PMA (200 nmol/L, 30 min), although no functional NMDA channels were detected after PMA-induced membrane trafficking. Immunostaining with an ER marker also revealed that NR2A subunits were exported from the ER after PMA treatment. Furthermore, the deletion of amino acids between 1149-1347 or 1354-1464 of NR2A inhibited PMA-induced surface expression of NR2A subunits. CONCLUSION: First, our data suggests that PMA treatment can induce the surface expression of homomeric NR2A subunits. Furthermore, this process is probably mediated by the NR2A C-terminal region between positions 1149 and 1464. PMID- 17112413 TI - Construction of a small peptide library related to inhibitor OM99-2 and its structure-activity relationship to beta-secretase. AB - AIM: To develop probes for detecting the binding specificity between beta secretase and substrate, and provide reliable biological activity data for further researching encircling substrate-based inhibitors. METHODS: To prepare the inhibitors, the hydroxyethylene (HE) segment including P1 and P1'was synthesized after multi-step reactions; the combination of all segments was then completed through solid phase synthesis. Recombinant human beta-secretase ectodomain (amino acid residues 1-460) was expressed as a secreted protein with a C-terminal His tag in insect cells using baculovirus infection, and all compounds were evaluated in this beta-secretase enzyme assay. In order to understand the interaction in detail, the theoretical methods, namely molecular dynamics (MD) simulation and molecular mechanics-generalized-born surface area (MM-GBSA) analysis, were performed on the complex of beta-secretase and OM99-2 to obtain the geometrical and energetical information. RESULTS: We designed and constructed a positional scanning combinatorial library including 16 compounds; all members of the library were synthesized based on HE dipeptide isostere. Structure activity relationship studies at the P4-P1 and P1' -P4'positions led to the discoveries of P and P'sides binding specificity and potent inhibitors 14, 18, and 22. The binding free energy on the whole system and every residue were compared to the biological assay result. CONCLUSION: The removal of P4' yielded inhibitor 22 (A3 *B2) with high potency; further truncation of P3'gave inhibitor 18 (A3 *B1) with equal activity, implying that the right side of the inhibitors play a less important role and could be easily simplified, while change on the P side may cause substantial results. PMID- 17112414 TI - CCR3 monoclonal antibody inhibits airway eosinophilic inflammation and mucus overproduction in a mouse model of asthma. AB - AIM: To explore the effect of a rat anti-mouse CC-chemokine receptor-3 (CCR3) monoclonal antibody (CCR3 mAb) on airway eosinophilia and mucus overproduction in asthmatic mice. METHODS: An asthma model was sensitized and challenged by ovalbumin (OVA) in male C57BL/6 mice. Asthmatic mice were given dual administration (intraperitoneal injection and aerosol inhalation) of CCR3 mAb or nonspecific rat IgG (ns-IgG). The number of total and differential inflammatory cells in the bronchial alveolar lavage fluid (BALF) was counted. Eosinophils number, the goblet cell percentage (GCP) and airway mucus index (AMI) were measured in the lung tissues. Interleukin (IL)-5 levels in the BALF were examined. The expression of MUC5AC and the epidermal growth factor receptor (EGFR) mRNA in the lung tissues was detected by semi-quantitative RT-PCR. The results were compared among the groups. RESULTS: CCR3 mAb significantly suppressed the increased eosinophils in the BALF and lung tissues in OVA challenged mice compared with ns-IgG-treated mice. IL-5 levels in the BALF in CCR3 mAb and ns-IgG administration mice exhibited no obvious changes relative to OVA-challenged asthmatic mice. CCR3 mAb reduced the increased GCP and AMI after OVA challenge and decreased the enhanced expression of MUC5AC and EGFR mRNA in lung tissues in asthmatic animals. CONCLUSION: CCR3 mAb can significantly inhibit airway eosinophilia and mucus overproduction in asthmatic mice. Blockage of CCR3 may represent a new strategy to asthma therapy. PMID- 17112415 TI - Green tea polyphenol epigallocatechin-3-gallate suppresses rat hepatic stellate cell invasion by inhibition of MMP-2 expression and its activation. AB - AIM: Epigallocatechin-3-gallate (EGCG) is the major component of green tea polyphenols, whose wide range of biological properties includes anti-fibrogenic activity. Matrix metalloproteinases (MMP) that participate in extracellular matrix degradation are involved in the development of hepatic fibrosis. The present study investigates whether EGCG inhibits activation of the major gelatinase matrix metalloproteinase-2 (MMP-2) in rat hepatic stellate cells (HSC). METHODS: The expression of MMP-2, tissue inhibitors of metalloproteinases 2 (TIMP-2), and membrane-type 1-MMP (MT1-MMP) was assessed by RT-PCR and Western blot analyses. MMP-2 activity was evaluated by zymography and MT1-MMP activity was assessed by an enzymatic assay. HSC migration was measured by a wound healing assay and cell invasion was performed using Transwell cell culture chambers. RESULTS: The expression of MMP-2 mRNA and protein in HSC was substantially reduced by EGCG treatment. EGCG treatment also reduced concanavalin A (ConA) induced activation of secreted MMP-2 and reduced MT1-MMP activity in a dose dependent manner. In addition, EGCG inhibited either HSC migration or invasion. CONCLUSION: The abilities of EGCG to suppress MMP-2 activation and HSC invasiveness suggest that EGCG may be useful in the treatment and prevention of hepatic fibrosis. PMID- 17112416 TI - Rat adipose-derived stromal cells expressing BMP4 induce ectopic bone formation in vitro and in vivo. AB - AIM: Bone morphogenetic protein 4 (BMP4) is one of the main local contributing factors in callus formation in the early phase of fracture healing. Adipose derived stromal cells (ADSC) are multipotent cells. The present study was conducted to investigate the osteogenic potential of ADSC when exposed to adenovirus containing BMP4 cDNA (Ad-BMP4). METHODS: ADSC were harvested from Sprague-Dawley rats. After exposure to Ad-BMP4, ADSC were assessed by alkaline phosphatase activity (ALP) assay, RT-PCR and von Kossa staining. BMP4 expression was assessed by RT-PCR, immunofluorescence and Western blot analysis. ADSC transduced with Ad-BMP4 were directly injected into the hind limb muscles of athymic mice. ADSC Ad-EGFP(enhanced green fluorescence protein) served as controls. All animals were examined by X-ray film and histological analysis. RESULTS: The expression of BMP4 was confirmed at both mRNA and protein levels. The expression of the osteoblastic gene, ALP activity and von Kossa staining confirmed that ADSC transduced with Ad-BMP4 underwent rapid and marked osteoblast differentiation, whereas ADSC transduced with Ad-EGFP and cells left alone displayed no osteogenic differentiation. X-ray and histological examination confirmed new bone formation in athymic mice transplanted with ADSC transduced with Ad-BMP4. CONCLUSION: Our data demonstrated successful osteogenic differentiation of ADSC transduced with Ad-BMP4 in vitro and in vivo. ADSC may be an ideal source of mesenchyme lineage stem cells for gene therapy and tissue engineering. PMID- 17112417 TI - (5R)-5-hydroxytriptolide inhibits IFN-gamma-related signaling. AB - AIM: (5R)-5-hydroxytriptolide (LLDT-8) displayed anti-arthritis and anti allogenic transplantation rejection activities in our previous studies. Here, we aim to further clarify the effect of LLDT-8 on the pro-inflammatory cytokine IFN gamma. METHODS: T cells were activated with anti-CD3 antibody or concanavalin A (ConA). The expression of cell surface molecules was detected with flow cytometry. Cells were labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) to test cell division. IFN-gamma production was determined by enzyme linked immunosorbent assay. Cell proliferation was evaluated by [3H]-thymidine uptake. Mice were immunized with ovalbumin to assess the in vivo immune response. RT-PCR and Real-time PCR were applied to determine the mRNA expression. The protein phosphorylation levels were detected by Western immunoblot assay. RESULTS: LLDT-8 at 100 nmol/L did not change the CD25, CD69, and CD154 expressions in anti-CD3-stimulated T cells. LLDT-8 markedly blocked the cell division of CD4 and CD8 T cells after ConA stimulation. LLDT-8 inhibited T cell derived IFN-gamma production. Moreover, LLDT-8 suppressed the ovalbumin-specific T cell proliferation and IFN-gamma generation. In anti-CD3-activated T cells, LLDT-8 abrogated the mRNA expression of signal transducer and activator of transcription1 (STAT1), T-box transcription factor, IL-12 receptor beta2, STAT4, and interferon regulatory factor 1 in the IFN-gamma expression pathway. Western blot analysis showed that LLDT-8 blocked the phosphorylation levels of extracellular signal-regulated kinase, stress-activated protein kinase (SAPK)/c Jun N-terminal kinase, and p38 mitogen-activated protein kinase in anti-CD3 plus anti-CD28-activated T cells. In addition, LLDT-8 reduced the transcripts of macrophage inflammatory protein (Mip)-1alpha, Mip-1beta, regulated upon activation normally T-cell expressed and secreted, inducible protein-10, IFN inducible T cell a chemoattractant, and monokine induced by IFN-gamma in IFN gamma-stimulated murine macrophage cell line Raw 264.7 cells. CONCLUSION: LLDT-8 was a potential inhibitor for IFN-gamma-associated signaling. PMID- 17112418 TI - Involvement of mitochondria and caspase pathways in N-demethyl-clarithromycin induced apoptosis in human cervical cancer HeLa cell. AB - AIM: To study the mechanisms by which N-demethyl-clarithromycin (NDC) induces human cervical cancer HeLa cell apoptosis in vitro. METHODS: The viability of N demethyl-clarithromycin-induced HeLa cells was measured by MTT assay. Apoptotic cells with condensed nuclei were visualized by phase contrast microscopy. Nucleosomal DNA fragmentation was assayed by agarose gel electrophoresis. Measurement of mitochondrial transmembrane potential was analyzed by a FACScan flowcytometer. Caspase-3, poly-(ADP-ribose) polymerase (PARP), caspase-activated DNase (ICAD), Bcl-2, Bax, p53, and SIRT1 protein expression and the release of cytochrome c were detected by Western blot analysis. RESULTS: N-demethyl clarithromycin, an anti-inflammatory substance, inhibited HeLa cell growth in a dose- and time-dependent manner. N-demethyl-clarithro-mycin induced HeLa cell death through the apoptotic pathways. The pan-caspase inhibitor (z-VAD-fmk), caspase-3 inhibitor (z-DEVD-fmk) and the caspase-9 inhibitor (z-LEHD-fmk) partially enhanced cell viability induced by N-demethyl-clarithromycin, but the caspase-8 inhibitor (z-IETD-fmk) had almost no effect. Caspase-3 was activated then followed by the degradation of caspase-3 substrates, the inhibitor of ICAD and PARP. Simultaneously, mitochondrial transmembrane potential was markedly reduced and the release of cytochrome c in the cytosol was increased. N-demethyl clarithromycin upregulated the expression ratio of mitochondrial Bax/Bcl-2, and significantly increased the expression of the p53 protein. It also downregulated anti-apoptotic protein SIRT1 expression. CONCLUSION: N-demethyl-clarithromycin induced apoptosis in HeLa cells via the mitochondrial pathway. PMID- 17112419 TI - Antisense oligonucleotides targeting midkine induced apoptosis and increased chemosensitivity in hepatocellular carcinoma cells. AB - AIM: Overexpression of midkine (MK) has been observed in many malignancies. This aim of this study is to screen for suitable antisense oligonucleotides (ASODN) targeting MK in hepatocellular carcinoma (HCC) cells and evaluate its antitumor activity. METHODS: Ten ASODN targeting MK were designed and synthesized. After transfection with ASODN, cell proliferation was analyzed with MTS[3-(4,5 dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H tetrazolium, inner salt] assay. In addition, MK mRNA, protein levels, as well as apoptosis and caspase-3 activity were also examined in HepG2 cells. Cell proliferation was then analyzed after treatment with both ASODN and chemotherapeutic drugs. RESULTS: In this experiment, the ASODN5 among the 10 ASODN showed higher inhibitory activity against proliferation of hepatocellular carcinoma cells in a dose-dependent manner. In HepG2 cells, ASODN5 could significantly reduce the MK mRNA level and protein content. After transfection with ASODN5 for 48 h, accompanied with a decline of survivin and Bcl-2 protein content, a remarkable increase of apoptosis and caspase-3 activity was observed in HepG2 cells. Furthermore, ASODN5 transfer can significantly increase chemosensitivity in HepG2 cells. CONCLUSION: Antisense oligonucleotides targeting MK shows therapeutic effects on HCC; ASODN5 has the possibility to be developed as an effective antitumor agent. PMID- 17112420 TI - Influence of intralipid on free propofol fraction assayed in human serum albumin solutions and human plasma. AB - AIM: It is generally assumed that only unbound drugs can reach the site of action by diffusing across the membranes and exerting pharmacological effects by interacting with receptors. Recent research has shown that the percentage of free drugs may depend on the total drug concentration. The aim of the paper is to verify whether the mentioned dependence reported for propofol also takes place in plasma and human serum albumin samples in the presence of intralipid-the medium used as a vehicle for propofol infusions and a parenteral nutrition agent. METHODS: Artificial plasma samples and human plasma were spiked with intralipid or ethanolic solutions of propofol. The samples were then assayed for free propofol concentration using ultrafiltration and high performance liquid chromatography with fluorimetric detection. RESULTS: The decrease of the total drug concentration results in free propofol fraction increase, irrespectively of the used type of propofol solvent and sample type. The addition of intralipid causes the lowering of the overall free drug fraction with respect to the samples spiked with ethanolic solutions of the drug. CONCLUSION: The presence of intralipid does not influence the phenomenon of free propofol fraction rise at low total drug concentration. Such a rise cannot be ignored in clinical conditions when the drug is applied for sedative, antiemetic or other low-dosage purposes. PMID- 17112421 TI - Pharmacokinetic behaviors and oral bioavailability of oridonin in rat plasma. AB - AIM: To study the intravenous and oral pharmacokinetic behavior of oridonin and its extent of absolute oral bioavailability in rats. METHODS: Oridonin was administered to rats via iv (5, 10 and 15 mg/kg), po (20, 40 and 80 mg/kg) or ip administration (10 mg/kg). The concentrations of oridonin in rat plasma were determined by a high performance liquid chromatography with electrospray ionization mass spectrometric detection (HPLC/ESI-MS) method and the pharmacokinetic parameters were determined by non-compartmental analysis. RESULTS: The plasma concentration of oridonin after intravenous administration decreased polyexponentially, and the pharmacokinetic parameters of oridonin were dose-independent within the examined range. Oridonin was absorbed rapidly after oral gavage with a t(max) of less than 15 min; the extent of absolute bioavailability of oridonin following oral administration was 4.32%, 4.58% and 10.8%. The extent of absolute bioavailability of oridonin following intraperitoneal administration was 12.6%. CONCLUSION: First order rate pharmacokinetics were observed for oridonin within the range of iv doses, while the extent of absolute oral bioavailability was rather low and dose- dependent. The low and dose-dependent extent of oral bioavailability may be due to the saturation of first-pass effects. PMID- 17112422 TI - Exercise for people with osteoporosis: translating the science into clinical practice. AB - The "holy grail" of inquiry regarding exercise and osteoporosis has been identifying a type of exercise that builds bone. Investigation using animal models has provided many insights into how bone responds to mechanical loading, but translating these findings into an exercise prescription for patients with osteoporosis is difficult. Patients expect bone to respond to exercise in a linear fashion, such as they are accustomed to experiencing with muscles in response to progressive strength training or with the cardiopulmonary system in response to endurance training. If the skeleton accrued greater mass in response to increasing intensity and duration of mechanical strain, our bones would weigh so much that we could not move. A unique requirement of bone is that adaptations to loading produce the strongest and the lightest structure. More exercise is not always better, but we are not yet sure exactly what and how much is enough and what and how much is too much. This complexity stymies clear communication, both in the clinic and in public health initiatives. PMID- 17112423 TI - Osteoporosis and depression: a historical perspective. AB - In the early 1980s, researchers studying osteoporosis noted that depression was one of the major negative consequences of bone loss and fractures. These researchers believed that osteoporosis and fractures occurred first, causing a reactive depression. Meanwhile, a similar but distinct psychiatry literature noted that osteoporosis or bone loss appeared to be an undesirable consequence of major depression. Here, depression was seen as the causal factor, and osteoporosis was the outcome. The psychiatric perspective is more biological, based on the presence of hypercorticoidism in depressed individuals. Those who believe that osteoporosis leads to depression point out that depression is a consequence of many chronic illnesses. Regardless of the correct causal order, the strong positive relationship between osteoporosis and depression merits further clinical and research attention in the future. PMID- 17112424 TI - Noninvasive assessment of bone microarchitecture by MRI. AB - In determining fracture risk, it has become apparent that bone mineral density accounts for only a portion of bone strength, with the remainder being determined by the material and structural properties of the bone tissue. Over the past 15 years, high-resolution MRI has provided a window into the structural nature of bone disease. Cross-sectional studies imaging the trabecular bone in patients with conditions ranging from postmenopausal osteoporosis to organ transplantation to renal osteodystrophy have all demonstrated a correlation of microarchitecture with fracture burden and have done so at a variety of anatomic sites. Recently, the utility of longitudinal studies for monitoring treatment in vivo has been demonstrated. This technique is noninvasive, involving no contrast or ionizing radiation, and provides useful clinical information independent of bone mineral density, thereby allowing for better classification of those at high risk for fracture. PMID- 17112425 TI - Prescreening tools to determine who needs DXA. AB - Clinical decision rules (CDRs) are designed to help physicians practice better. A number of CDRs to assist in identifying women with low bone mass have been developed since the mid 1990s, including SCORE, OST (OSTA), OSIRIS, SOFSURF, NOF, ABONE, pBW, ORAI, and weight-only-EPIDOS (which we have termed WO-E). This review discusses these CDRs in terms of development and validation cohorts and their sensitivity and specificity. The sensitivities of the available CDRs exceed 80% and specificities are about 50%. After much analysis, it appears that most experts prefer OST for its simplicity and SCORE for its flexibility, but there is no consensus on what risk factors to use in the CDRs and what regions of interest (spine, total hip, femoral neck, or a combination) to test with dual-energy x-ray absorptiometry (DXA). Because of the lack of consensus, there are barriers to the clinical application of these CDRs. Agreement on a single CDR for worldwide use is required to optimally fulfill the objective of identifying low bone mass. PMID- 17112426 TI - Assessing bone mass in children and adolescents. AB - Growing awareness that osteoporosis may have its antecedents in childhood has led to increasing interest in assessing bone mass in children and adolescents. Several noninvasive imaging techniques are currently available to measure properties of the growing skeleton, including bone mass, density, cross-sectional area, and microarchitecture. Dual-energy x-ray absorptiometry (DXA) is the most widely used technique, but it has several major limitations associated with its dependence on two-dimensional projections. Quantitative CT and peripheral quantitative CT allow three-dimensional imaging but are more costly and have higher radiation exposure. Quantitative ultrasound is simple and inexpensive but can measure bone "quality" only at a single peripheral site. MRI techniques for measuring bone are still under development and not yet ready for clinical use. For all of these techniques, clinical interpretation of the bone measures obtained remains a significant challenge. Further research is needed to relate these measures to osteoporosis in the elderly and to short-term and long-term fracture risk. PMID- 17112427 TI - Treatment of children with osteogenesis imperfecta. AB - Children with moderate to severe forms of osteogenesis imperfecta (OI) require adequate physiotherapy, rehabilitation and orthopedic surgery. Supportive treatment with bisphosphonates can improve the effects of these nonmedicinal treatment modalities. Benefits of bisphosphonate treatment include decreased pain, lower fracture incidence, and better mobility. Among the various bisphosphonates, intravenous pamidronate has been studied in most detail. However, the optimal treatment regimen and the long-term consequences of pamidronate treatment in children are currently unknown. Given these uncertainties, treatment with bisphosphonates during growth should be reserved for patients who have significant clinical problems, such as vertebral compression fractures or long-bone deformities. Medical therapies other than bisphosphonates play a minor role at present. Gene-based therapy currently remains in the realm of preclinical research. PMID- 17112428 TI - Where are we going with chlamydia? PMID- 17112429 TI - The use of focus groups to design an internet-based program for chlamydia screening with self-administered vaginal swabs: what women want. AB - OBJECTIVE: To ascertain the opinions, concerns and perceptions of sexually active women to guide the development of an internet-based chlamydia outreach and screening program using self-administered vaginal swabs as a first step to prevention. METHODS: Seven focus groups were conducted by trained facilitators. Questions were designed to initially open the discussion and elicit the members' own perceptions. Secondary, more probing questions were asked later to confirm participants' responses and elicit truthful answers. The main discussion topics were women's ideas about internet recruitment for chlamydia screening, preferred genital sample type, self-sampling at home using vaginal swabs and using the mail to return specimens. Participants were 42 women, aged 14-49 years. Structured discussions were facilitated using open-ended questions about access to chlamydia testing via the internet. Data were collected and reviewed for common themes and emphasis. RESULTS: All women actively participated in the discussions, providing valuable information. The concepts of self-sampling and the overall project were viewed positively, along with draft advertisements, questionnaires and self sampling instructions; some modifications were suggested. Common themes included offering free kits available within their community or by direct mail, as well as pre-addressed, stamped mailers for returning the kit to the laboratory for testing. Commonly perceived obstacles and potential risks included: maintenance of confidentiality; situations of embarrassment; and ensuring simplicity of packaging. Women indicated confidence in their ability to collect vaginal specimens and willingness to call for their test results. CONCLUSIONS: Focus group surveys were a useful tool and provided valuable feedback to inform the design of a specialised website to educate and facilitate access to chlamydia screening through home sampling. PMID- 17112430 TI - A survey of partner notification practices among general practitioners and their use of an internet resource for partner notification for Chlamydia trachomatis. AB - OBJECTIVES: To determine in which circumstances Victorian general practitioners (GPs) offer chlamydia testing to patients, the attitudes of GPs in relation to contact tracing, how often GPs use a pre-printed partner letter and patient brochure and what proportion of GPs have immediate internet access in their consulting rooms. METHODS: This study involved two parts, an initial survey of a sample of GPs in Victoria and a study of GP use of a website that provided treatment guidelines, a printable client brochure and a partner letter. RESULTS: Of 418 eligible GPs, 221 (53%, 48-58%, 95% CI) returned completed surveys. Of these, 213 (97%, 93-99%, 95% CI) GPs believed that patients were largely responsible for notifying partners. Partner letters were rarely used: 167 (76%, 70-81%, 95% CI) GPs reported they never used partner letters, 18 (8%, 5-13%, 95% CI) GPs reported rare use and 23 (10%, 7-15%, 95% CI) GPs reportedusing them sometimes. Of the GPs, 181 (82%, 77-87%, 95% CI) reported they would find a partner letter and patient brochure on a website helpful. During the study, the website was accessed by 28 GPs (25%, 17-34%, 95% CI) in Gippsland and 17 GPs (8%, 5-13%, 95% CI) in Geelong who received positive chlamydia results on 110 and 208 clients respectively. CONCLUSIONS: GPs mostly considered patients responsible for partner notification but uncommonly used partner letters or an information brochure to assist them. Importantly, GPs reported that they could improve partner notification if further support was provided. In addition, when a website was provided with useful documents on it, up to 25% of GPs used it. This indicates that simple and inexpensive interventions can support GPs with strategies that may improve the control of chlamydia. PMID- 17112431 TI - Antibiotic consumption and chlamydia prevalence in international studies. AB - BACKGROUND: To determine whether there is an ecological association between antibiotic use and chlamydia prevalence. METHODS: A systematic review was undertaken of international studies on chlamydia prevalence among women aged 15 25 years published between 2000 and 2005. Preference was given to studies using nucleic acid testing and representative population-based sampling methods. Data were obtained on per capita antibiotic consumption according to the defined daily dose. RESULTS: For the 12 countries for which both antibiotic consumption and relevant prevalence data for chlamydia were available, a non-significant negative correlation was found between total antibiotic consumption per capita and chlamydia prevalence among younger women according to country (r(s) = -0.242, P = 0.449). When an outlier (from the Netherlands) was excluded, the correlation was significant (r(s) = -0.615, P = 0.044). Combined use of tetracyclines and macrolides was also associated with lower chlamydia prevalence (r(s) = -0.697, P = 0.017). CONCLUSIONS: It is possible that antibiotics used for other reasons may have unexpectedly reduced the prevalence of chlamydia. PMID- 17112432 TI - The cost effectiveness of screening for genital Chlamydia trachomatis infection in Australia. AB - BACKGROUND: In Australia, there is no published study on the cost effectiveness of screening for chlamydia. The aim of this study was to examine the cost effectiveness of a hypothetical screening programme for chlamydia based on annual opportunistic testing of all women 25 years of age or younger consulting a general practitioner, compared with no screening. METHODS: A decision-analytic modelling approach was used to determine the incremental cost effectiveness ratio (ICER) of screening compared with no screening over 25 years. The analysis measured Australian health-care costs and benefits were assessed in terms of quality-adjusted life years (QALYs). RESULTS: The analysis resulted in a cost per QALY of 2968 dollars for screening. One-way sensitivity analyses on all variables, and multi-way sensitivity analyses on some variables, showed a wide range for the cost effectiveness, from dominance (where screening is effective and saves money overall) to an ICER of 67,715 dollars per QALY. CONCLUSIONS: The results indicate that annual opportunistic screening for chlamydia in women under 25 is a potentially worthwhile undertaking. However, the analysis also highlights uncertainties around the natural history of chlamydia and the effectiveness of chlamydia screening. Given these uncertainties, the need for further primary data collection in these areas becomes apparent. PMID- 17112433 TI - A chlamydia prevalence survey of young women living in Melbourne, Victoria. AB - BACKGROUND: To estimate the population-based chlamydia prevalence among women aged 18 to 35 years living in Melbourne, Victoria, and to assess the feasibility of using mailed urine specimens to test women. METHODS: A simple random sample of 11,001 households in Melbourne was selected from the telephone directory. Participants completed telephone interviews and provided urine specimens through the mail for chlamydia testing. Urines were tested using polymerase chain reaction. RESULTS: 11,001 households were contacted, with 1532 households identified as including eligible women; telephone interviews were completed, with 979 women giving a response rate of 64%. Six hundred and fifty-seven women provided a urine specimen with a response rate of 43%. Among sexually active women aged 18-24 years, the chlamydia prevalence was 3.7% (95% CI: 1.2%, 8.4%) and 0.2% (95% CI: 0.0%, 1.1%) among 25-35 year olds. Chlamydia prevalence increased significantly with an increasing number of male sexual partners. CONCLUSIONS: This is the first study of its kind in Australia and shows that chlamydia prevalence increases with an increasing number of male sexual partners in the last 12 months. Mailed urine specimens are feasible for conducting population-based chlamydia-prevalence surveys but it is difficult to obtain high response rates with this methodology. Public health resources should now be directed towards investigating how to reach young women at increased risk of infection, ensuring that they are tested for chlamydia. PMID- 17112434 TI - Chlamydia testing in general practice - a survey of Victorian general practitioners. AB - BACKGROUND: To assess the current chlamydia testing practices of Victorian general practitioners (GPs). METHODS: GPs were randomly selected from the Australasian Medical Publishing Company's national database of medical practitioners and mailed a letter of invitation asking them to complete a postal survey. Up to three postal reminders were sent to non-responders. RESULTS: Of 421 eligible GPs, 252 (60%) returned a completed survey; 22.9% (95% CI: 17.8%, 28.6%) reported testing at least some asymptomatic patients for chlamydia each week and 26.8% (95% CI: 21.4%, 32.7%) reported that they presumptively treated patients for chlamydia without testing them at least half the time. The majority knew the appropriate specimens for diagnosing chlamydia, but 6-8% thought blood and 6% indicated that the Pap smear could be used to reliably diagnose chlamydia infection. CONCLUSIONS: These findings have implications for the future chlamydia screening pilot program in Australia and indicate that a comprehensive education program will be necessary to inform GPs and equip them with the skills to appropriately test for chlamydia in their practice. PMID- 17112435 TI - Comparison of self-reported and test-identified chlamydial infections among young adults in the United States of America. AB - BACKGROUND: Many studies rely on respondent reports of prior diagnosed sexually transmissible infections (STIs), but these self reports are likely to under estimate infection prevalence. The extent of bias from using self-reported STI data, and whether bias varies by sex and race, is largely unknown. This gap is addressed using a large, nationally representative sample. METHODS: Cross sectional analyses of Wave III of the National Longitudinal Study of Adolescent Health. Participants were 18-26 years old (n = 12,359). Estimates of the prevalence of chlamydial infection based on self-reported diagnoses in the past year were compared with actual prevalence based on nucleic acid amplification testing (NAAT) at the time of data collection. Ratios of test-identified prevalence to self-reported diagnosis prevalence were calculated by sex and race/ethnicity groups. Larger ratios indicate greater extent of self reports under-estimating infection prevalence. RESULTS: About 4.2% of the sample had a current NAAT-identified chlamydial infection, but only 3.0% reported having been diagnosed with chlamydia in the past year, yielding a ratio of 1.43. The ratio of test-identified infection prevalence to prevalence identified from self-reported diagnoses was larger among men than women (2.07 versus 1.14, P < 0.05). Among men, the ratio was larger among non-Hispanic blacks (2.40) compared with non Hispanic whites (1.07, P < 0.05). CONCLUSIONS: Use of self-reported diagnoses under-estimates chlamydial infection prevalence, particularly among men, and among non-Hispanic black men. Reliance on self-reported STIs may consequently lead to biased conclusions, particularly for these groups. Use of biological testing for STIs in research studies is recommended. PMID- 17112436 TI - Prevalence of Chlamydia trachomatis infection among 'high risk' young people in New South Wales. AB - International research on homeless adolescents has found that incidence and prevalence of sexually transmissible infections is relatively high. This study reports on a chlamydia prevalence survey conducted among high-risk young people (14-25 years) in New South Wales. The participants were recruited from youth health centres, which target homeless and high-risk youth. Of 333 clients (42.6% male), 84.1% were sexually active and mean number of sexual partners over the preceding 3 months was 1.4. Among sexually active participants, 24.6% claimed to use condoms always and 25% never. Sixteen of 274 available urine samples tested positive for Chlamydia trachomatis infection. Further research is warranted to better define high-risk groups and clarify the nature of associations between various factors impacting on sexual health. Most importantly, research is now called for into effective strategies for engaging and attracting young people to screening, treatment and contact tracing. PMID- 17112437 TI - Erection loss in association with condom use among young men attending a public STI clinic: potential correlates and implications for risk behaviour. AB - BACKGROUND: To assess prevalence of condom-associated erection loss and to identify correlates of erection loss among men attending a sexually transmissible infections (STI) clinic. METHODS: Men (n = 278) attending an STI clinic responded to an anonymous questionnaire aided by a CD recording of the questions. The sample was screened to include only men who had used a condom during penile vaginal sex at least three times in the past 3 months. Erection loss was assessed for 'the last three times a condom was used'. RESULTS: The mean age of the participants was 23.7 years (s.d. = 4.1); 37.1% of the men reported condom associated erection loss on at least one occasion. Men who had reported condom associated erection loss were also reported having more frequent unprotected vaginal sex (P = 0.04) and were less likely to use condoms consistently (P = 0.014) than men without erection loss. Men with erection loss were also more likely to remove condoms before sex was over (P = 0.001). Age and race/ethnicity were not associated with erection loss. In multivariate analysis, three significant statistical predictors were identified: low self-efficacy to use condoms (P = 0.001); problems with 'fit or feel' of condoms (P = 0.005); and having more than three sex partners during the previous 3 months (P = 0.02). CONCLUSIONS: Condom-associated erection loss may be common among men at risk for STIs. This problem may lead to incomplete or inconsistent condom use. Men may be more likely to experience condom-associated erection loss if they lack confidence to use condoms correctly, if they experience problems with the way condoms fit or feel, and if they have sex with multiple partners. PMID- 17112438 TI - Sex preparation and diaphragm acceptability in sex work in Nairobi, Kenya. AB - BACKGROUND: Women in sex work stand to benefit if the contraceptive diaphragm alone or combined with a microbicide proves to be an effective barrier method against HIV and sexually transmissible infection (STI). Currently, contraceptive diaphragm users are advised to leave the diaphragm in situ without concomitant use of other intravaginal substances for at least 6 h after intercourse. METHODS: We conducted in-depth interviews on sexual behaviour including post-coital intravaginal practices with 36 women in sex work and 26 of their clients and held two focus-group discussions, each with 10 women. RESULTS: The women described adapting several potentially harmful substances, such as cloth and soapy water, for post-coital vaginal use to ensure personal hygiene, disease prevention and client pleasure. Some wanted to clean themselves and remove the diaphragm early, fearing exposure to HIV infection for themselves and their subsequent clients. Clients indicated their desire for 'dry sex', vaginal cleanliness and reduced risk of infection through vaginal cleaning. CONCLUSIONS: The diaphragm as a female-controlled barrier method for HIV/STI prevention may have limited acceptability among women in sex work if its effectiveness depends on a 6-h post coital wait before removal, along with avoidance of concomitant use of intravaginal substances. In keeping with the beliefs of the the female sex workers and their needs and practices, alternative intravaginal substances and modes of insertion that will not disrupt vaginal flora, injure vaginal epithelium, damage the diaphragm or counteract potentially beneficial effects of microbicides are needed. The possibility of removing the diaphragm sooner than the recommended 6 h for contraception should be further studied. PMID- 17112439 TI - Seroprevalence of herpes simplex virus-1 and -2 in attendees of a sexually transmitted infection clinic in Singapore. AB - BACKGROUND: We studied the seroprevalance of HSV (herpes simplex virus)-1 and HSV 2 in outpatient attendees of a sexually transmitted infection (STI) clinic in Singapore and examined their knowledge and attitudes towards HSV infection. METHODS: Two hundred male and 200 female participants were recruited in the study. Questionnaires were administered and blood samples were taken and analysed using the HerpeSelect 1 and 2 enzyme-linked immunosorbent assay (ELISA) IgG assays (type-specific serological tests). RESULTS: HSV-1 was positive in 223 (55.8%) individuals, negative in 175 (43.8%) and indeterminate in two (0.5%), whereas HSV-2 was positive in 114 (28.5%) individuals, negative in 284 (71.0%) and indeterminate in two (0.5%). The seroprevalance of HSV-2 was 26% and 31% in males and females, respectively. The correlation between a previous history of cold sores and HSV-1 infection was poor. On univariate analysis, there was a significant association with age, marital status and years of sex (P < 0.05) but after adjusting for confounders, none of the variables were significantly associated with HSV-2 seroprevalance. Most of the respondents (65.8%) were aware that herpes is an STI, whereas only half of them were aware of the possibility of asymptomatic transmission. CONCLUSION: Although HSV-2 is a common infection among STI clinic attendees in Singapore, there is an unsatisfactory level of knowledge among the attendees about HSV infection and public education programs should be introduced to address this. PMID- 17112440 TI - Prevalence and risk factors for lifetime exposure to Pap smear abnormalities in the Australian community. AB - BACKGROUND: This study examined the prevalence, correlates and consequences of abnormal Pap smears in a population-based survey of sexuality and health in the Australian community. METHODS: Cross-sectional telephone survey of 908 women aged 18-59 years randomly selected from the Commonwealth electoral roll. RESULTS: Most women (91%) reported having had at least one Pap smear test, a figure directly comparable with national estimates. Being single (prevalence ratio (PR) 4.61; 95% CI 2.09-10.17) and not having had sexual intercourse (PR 5.31, 95% CI 3.11-9.07) were strong predictors of never having been tested. One in four women (26%) who reported being screened also reported having had an abnormal Pap smear result, of whom 66% said they had further testing and 52% some form of treatment. A minority (19%) reported negative effects of treatment on their sex lives. Having been diagnosed with human papillomavirus (HPV) (PR 2.87, 95% CI 1.84-4.48), and to a lesser degree, having had a greater number of male sexual partners (PR 1.38, 95% CI 1.01-1.89), and experiencing sexual problems in the last year (PR 0.99, 95% CI 0.99-1.88) were independently associated with reporting of abnormal Pap smear results. CONCLUSIONS: Approximately one in four women self report lifetime exposure to Pap smear abnormalities. It is important that women are well prepared for this common experience. A causal association between multiple sexual partners and risk of acquiring HPV infection is supported by these data. PMID- 17112441 TI - Cysteine 138 mutation in HIV-1 Nef from patients with delayed disease progression. AB - BACKGROUND: The nef gene from HIV-1 has been shown to be an important pathogenic factor when considering development of AIDS. Detection of nef variants with an effect on immune modulation is important to understand HIV-1 pathogenesis and has possible impact on treatment strategies. METHODS: The nef gene of HIV-1 isolates from patients in a long-term non-progressor (LTNP) cohort and a slow-progressor (SP) cohort (n = 11) was analysed and compared with isolates from a control patient group of progressors (n = 18). Most of the patients with delayed disease progression had extensive medical records, providing an insight into the LTNP disease profile and allowing for the stratification of patients based on their CD4 cell decline. RESULTS: In sequences from nine patients, most of the functional domains of HIV-1 Nef appeared intact, and no major deletions were observed to possibly account for an effect on the delayed disease status. However, the results demonstrate a high incidence of a single amino acid polymorphism (cysteine 138) in HIV-1 Nef. The allelic frequency of cysteine 138 between the delayed disease progression group and the progressor group was found to be statistically significant (P = 0.0139). The phylogeny of isolates was investigated and the variants harbouring the cysteine 138 mutation clustered independently. CONCLUSION: The present study describes a viral genetic polymorphism related to AIDS disease progression. The polymorphism (cysteine 138) has previously been reported to confer decreased viral replication (Premkumar DR, et al. AIDS Res Hum Retroviruses 1996; 12(4): 337-45). A sequence database search for comparative mutations revealed a high frequency of cysteine 138 in patients with reported SP AIDS. PMID- 17112442 TI - A randomised controlled trial of omega-3 fatty acid supplementation for the treatment of hypertriglyceridemia in HIV-infected males on highly active antiretroviral therapy. AB - BACKGROUND: Hypertriglyceridaemia is a recognised metabolic abnormality in HIV infected people, increasing in severity in people treated with highly active antiretroviral therapy (HAART). An alternative treatment for hypertriglyceridaemia in non-HIV-infected populations is omega-3 fatty acid supplementation. This study aimed to compare the effectiveness of omega-3 fatty acid supplementation and placebo in lowering fasting triglyceride levels in HIV infected patients on HAART. METHODS: A placebo-controlled, randomised, double blind trial in participants on stable HAART with fasting triglycerides of >3.5 mm to 10.0 mm using 9 g of omega-3 fatty acids versus placebo (olive oil) after a 6 week lead in on dietary therapy. RESULTS: Eleven patients were enrolled. The mean triglyceride level for the population decreased from 5.02 mm at baseline to 4.44 mm (-11.6%) after dietary intervention and 3.37 mm (-32.9%) after the 8-week treatment period. In the omega-3 fatty acid arm of the study, triglycerides fell from 5.34 mm to 5.02 mm (-6%) after dietary intervention and to 2.30 mm (-56.9%) after the treatment period. In the placebo arm of the study, triglycerides fell from 4.77 mm to 4.05 mm (-15.1%) after dietary intervention and to 4.08 mm ( 14.5%) after the treatment period. Using the random effects model, a statistically significant effect on triglycerides of omega-3 fatty acid versus placebo was found (chi(2) = 6.04, P = 0.0487). The estimated difference between groups for change in mean triglycerides over 8 weeks was -2.32 mm (95% CI -4.52, 0.12 mm). CONCLUSIONS: Omega-3 fatty acids are likely to be an effective treatment for hypertriglyceridaemia in HIV-infected males on HAART. PMID- 17112443 TI - Delayed diagnosis of HIV: missed opportunities and triggers for testing in the Australian Capital Territory. AB - BACKGROUND: To determine (i) the rate of delayed HIV diagnosis; (ii) the missed opportunities for HIV diagnosis; and (iii) to identify who initiates HIV testing and what triggers them to do so. METHODS: An analysis of the case records of all HIV-positive patients who attended Canberra Sexual Health Centre (CSHC) between 1985 and 2005 was conducted. RESULTS: During the study period, 319/355 CSHC patients diagnosed with HIV had sufficient data to allow analysis regarding the timeliness of their diagnosis. Of these, 52 (16.3%) received a delayed diagnosis. The rate of delayed diagnosis was 9.7% (95% CI 5.1-15.3) in the 1980s and 25.6% (95% CI 13-42.1) between 2000 and 2004. There were no statistically significant differences in sociodemographic or behavioural characteristics between patients with delayed and timely HIV diagnoses. To determine who initiated testing, and if there were missed opportunities for testing, the records of CSHC patients diagnosed with HIV between 1995 and 2005 were examined. Of the 115 people diagnosed in this period, only 71 had documentation concerning missed opportunities for testing. Forty-one of these (58%) had been in contact with a health professional while infected, but before their diagnosis of HIV and 39/41 (95%) had a significant risk factor in their history that could have initiated an HIV test. Clinicians initiated testing for 43.5% of the patients, 11.3% were identified through contact tracing and only 28.7% were self referred for testing. CONCLUSIONS: Late diagnosis of HIV is common in the Australian Capital Territory and may have increased over time. Clinicians need to be aware of the sometimes subtle manifestations of early and late HIV infection and have a lower threshold for HIV antibody testing. PMID- 17112444 TI - Bacterial vaginosis in women of low socioeconomic status living in slum areas in Chennai, India. AB - Bacterial vaginosis (BV) is a common cause of vaginitis among women of childbearing age. This study was performed to investigate the prevalence of BV and its association with sexually transmitted infections among 487 women of low socioeconomic status. Blood, vaginal and endocervical swabs were tested for HIV, herpes simplex virus-2 (HSV-2), Treponema pallidum, BV, Chlamydia trachomatis, Neisseria gonorrhoea and Trichomonas vaginalis. Of the women screened for BV, 120 (25, 95% CI 20.8-28.4) were positive and 40 (8.2, 95% CI 5.8-10.6) were intermediate. Bacterial vaginosis was significantly associated with age >25 (P = 0.014) and sexual experience (P = 0.085). Bacterial vaginosis was also related to concurrent infections with T. vaginalis (relative risk (RR) = 6.6, 95% CI 2.8 15.5, P = 0.000) and HSV-2 (RR = 2, 95% CI 1.3-2.9; P = 0.0031). The role of other possible risk factors needs to be explored. PMID- 17112445 TI - Human papillomavirus prevalence in Canberra high school students: significance for vaccination strategies and adolescent health. PMID- 17112446 TI - Sexually transmissible infections among illegal female sex workers in Israel. AB - Due to the mobile and clandestine nature of those who enter a country illegally, female sex workers (FSWs) who are working without papers or work permits often have no access to sexual health care. This study reports on the sexually transmissible infection (STI) prevalence among a sample of 43 sex workers working illegally. Brothel workers from republics of the Former Soviet Union (FSU), working in two locales in Israel were tested for the presence of eight pathogens and the presence of pathology by Pap smear. Of these brothel workers, 48.8% had at least one positive STI result, 14% had two STIs and one woman had three STIs. There were no cases of HIV, gonorrhoea or malignancy detected; high rates of ureaplasma (26.8%) and chlamydia were found (16.7%). Four cases of hepatitis C (9%) and three cases of hepatitis B (7%) and mycoplasma (7%) were detected. There was no relationship between reported symptoms and the detection of STIs. The level of STIs is high among this population of FSWs and it is imperative to develop more accessible health services for these women. PMID- 17112447 TI - Adjuvant endocrine therapy in hormone receptor-positive postmenopausal breast cancer: evolution of NCCN, ASCO, and St Gallen recommendations. AB - Endocrine therapy has a firm role in adjuvant treatment of women with hormone receptor-positive invasive breast cancer. Until recently, tamoxifen was the most commonly used adjuvant endocrine therapy in premenopausal and postmenopausal women. Several randomized clinical trials have studied the third-generation selective aromatase inhibitors (AIs) (anastrozole, letrozole, and exemestane) as adjuvant endocrine therapy in postmenopausal women. These studies compared therapy with an AI alone versus tamoxifen alone; 2 to 3 years of tamoxifen followed by switching to an AI versus continuation of tamoxifen; or extended therapy with an AI after approximately 5 years of tamoxifen therapy. No statistically significant differences in overall survival were observed. A single trial using extended treatment with an adjuvant AI suggests a small, statistically significant survival advantage in women with axillary lymph node positive disease while showing no statistically significant decrease in survival with the use of an AI. The toxicities of the AIs are generally acceptable, with fewer endometrial cancers, gynecologic complaints, and thromboembolic events, but more bone fractures and arthralgias compared with tamoxifen alone. Three widely disseminated treatment guidelines, the National Comprehensive Cancer Network Breast Cancer Clinical Practice Guidelines in Oncology, the American Society of Clinical Oncology Technology Assessment on the Use of Aromatase Inhibitors, and the St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer, now incorporate AIs in the adjuvant therapy of postmenopausal women with estrogen receptor-positive breast cancer. PMID- 17112448 TI - Bladder cancer. Clinical guidelines in oncology. PMID- 17112449 TI - Surgical management of upper tract transitional cell carcinoma. AB - Upper tract transitional cell carcinoma (TCC) accounts for approximately 5% of urothelial tumors. Surgical therapy for upper tract TCC is based on tumor grade, stage, location, and confounding factors of individual cases. Options for treatment range from minimally invasive procedures, such as ureteroscopy, to open nephroureterectomy. Laparoscopic nephroureterectomy is progressively eclipsing open nephroureterectomy in the surgical management of upper tract TCC. This article discusses the surgical options for managing upper tract TCC and their considerations for use. PMID- 17112450 TI - Importance of node dissection in relation to neoadjuvant and adjuvant therapy. AB - Since the advent of effective chemotherapeutic regimens for treating transitional cell carcinoma, multimodal therapy has become part of the contemporary management of patients with muscle-invasive bladder cancer. However, radical cystectomy with pelvic lymphadenectomy remains the cornerstone of treatment for patients with localized and regionally advanced muscle-invasive disease. The effectiveness of chemotherapy models in bladder cancer can depend greatly on the quality of surgery. Unfortunately, without sufficient level I data, the boundaries of lymphadenectomy and the diagnostic and therapeutic ramifications of variations in the pelvic lymph node dissection remain undetermined. This article examines the role of pelvic lymph node dissection during perioperative chemotherapy and discusses the current challenges in establishing standards for lymphadenectomy in patients undergoing treatment for muscle-invasive bladder cancer. PMID- 17112451 TI - Contemporary intravesical treatment options for urothelial carcinoma of the bladder. AB - To provide a comprehensive review of intravesical treatment options for non muscle-invasive bladder cancer, we performed a search of the PubMed database for articles between 1980 and 2006 that reported on intravesical agents for treating this disease. Data were compiled and analyzed, emphasizing findings from large multicenter trials, studies providing reproducible results, data that could be confirmed by cross-referencing the literature, and phase I or II studies for pertinent novel agents. A critical analysis of evidence shows that: 1) treatment with Bacillus Calmette-Guerin (BCG), including a maintenance schedule (with or without interferon-alpha), is the most effective therapy for limiting recurrence, is the only therapy that reduces the incidence of progression, and overall is superior to chemotherapy; 2) mitomycin C, gemcitabine, anthracyclines, and thiotepa provide similar benefits for preventing recurrence in patients with minimal effect on progression; and 3) using chemotherapeutic agents immediately after transurethral resection (when use of BCG is contraindicated because of the risk for systemic absorption) reduces the recurrence rate by up to 50% and seems to be the ideal method of chemotherapy. Although various clinical factors dictate which agent is most appropriate for an individual patient, the current literature supports a single perioperative dose of intravesical mitomycin C followed, in appropriate cases, by induction and maintenance therapy with intravesical BCG. PMID- 17112452 TI - Testicular cancer. Clinical practice guidelines in oncology. PMID- 17112453 TI - Review of late complications of treatment and late relapse in testicular cancer. AB - With testicular cancer, a disease with a cure rate of 95%, the challenge is to restore quality of life to pretreatment levels and sustain it long-term. Although the implementation of guidelines and optimization of treatment modalities over the past years have served this purpose, some complications remain inevitable and experts are still challenged with late complications of outdated treatment standards. This article focuses on the late complications of cisplatin-based chemotherapy without disregarding those of currently applied infradiaphragmatic radiation. The most serious long-term complications of chemotherapy or radiotherapy are cardiovascular toxicity and second malignancies, as each has a 25-year risk of approximately 16%. Compared with the general population, risk for second malignancies remains significantly increased for at least 35 years after treatment. Chemotherapy-related cardiovascular toxicity is probably a result of both direct endothelial damage induced by cisplatin and indirect hormonal and metabolic changes. The increased incidence of the metabolic syndrome identified in long-term survivors is most likely associated with the lower testosterone levels reported. Cisplatin-based chemotherapy affects not only Leydig cells but also Sertoli and germ cells, resulting in infertility in 30% to 50% of testicular cancer patients treated with chemotherapy. Chronic neurotoxicity occurs in half of men, whereas permanent ototoxicity and some degree of renal function impairment occur in up to 30%. Pulmonary fibrosis, occurring in 5% to 10% of patients treated with bleomycin, is fatal in 1%. Although current treatment of advanced disease has changed its natural course, we are challenged by an increasing incidence of late relapse, an entity with a distinct tumor biology characterized by latency and chemoresistance. PMID- 17112454 TI - Kidney cancer. Clinical practice guidelines in oncology. PMID- 17112455 TI - Optimal management of small renal masses. AB - The incidence of small renal masses is increasing. Limited data suggest that although most are malignant, they are also likely to show indolent behavior. The tendency of these tumors to occur in older patient populations with multiple comorbidities raises important issues. Retrospective and prospective studies suggest that a substantial proportion of these small renal masses can be observed without compromising the opportunity for cure or subjecting patients to invasive procedures that might impact negatively on quality of life. Less-invasive treatment modalities, including partial nephrectomy and energy ablation techniques, provide multiple options for some patients. This article reviews selected aspects of the management of small renal masses, including their natural history, imaging, diagnostic biopsies, and treatment options. PMID- 17112456 TI - Understanding children's injury-risk behavior: wearing safety gear can lead to increased risk taking. AB - The present study examined whether school-age children show risk compensation and engage in greater risk taking when wearing safety gear compared to when not doing so when running an obstacle course containing hazards that could lead to physical injury. Because sensation seeking has been shown to influence risk taking, this child attribute was also assessed and related to risk compensation. Children 7-12 years of age were videotaped navigating the obstacle course twice, once wearing safety gear and once without safety gear, with reverse directions used to minimize possible practice effects. The time it took the child to run through the course and the number of reckless behaviors (e.g., falls, trips, bumping into things) that the child made while running the course were compared for the gear and no-gear conditions. Results indicated that children went more quickly and behaved more recklessly when wearing safety gear than when not wearing gear, providing evidence of risk compensation. Moreover, those high in sensation seeking showed greater risk compensation compared with other children. Implications for childhood injury prevention are discussed. PMID- 17112457 TI - [The prevention of intestinal obstruction related to adhesions]. PMID- 17112459 TI - Insights into the bilateral cortical control of human masticatory muscles revealed by transcranial magnetic stimulation. AB - In this brief review I describe details of the functional organisation of the bilateral corticobulbar projections to the trigeminally innervated masticatory muscles, as revealed by transcranial magnetic stimulation of the human brain. The motor cortices of both hemispheres are involved in control of trigeminal motoneurons, however the contralateral hemisphere has the greater excitatory influence. Corticomotoneuronal cells in each hemisphere project to jaw-closer and jaw-opener motoneurons. Less is known about cortically mediated inhibitory effects in the trigeminal motor system, but the available evidence suggests that drive to jaw muscles on each side is affected similarly by intracortical inhibitory processes activated in one hemisphere. Functional studies reveal that the two hemispheres play distinct roles in control of ipsilateral and contralateral muscles, particularly for jaw-closers. Masseter and digastric motor units recruited during low-force contractions do not receive uniform inputs from each hemisphere; the majority of masseter motor units are excited only from the contralateral hemisphere, and while digastric motor units are usually excited from both hemispheres the direct CM cell influence appears to be augmented on the contralateral side by corticobulbar activation of segmental excitatory interneurons. Differences in bilateral cortical control of jaw-closer and jaw opener muscles may contribute to the more independent control of jaw-closers on each side during functional tasks. Corticobulbar control of the trigeminal muscles during natural tasks such as chewing and speech remains to be investigated with TMS. PMID- 17112458 TI - An evaluation of backpack harness systems in non-neutral torso postures. AB - Much of the research on backpack design has been focused on spinal loading/biomechanics while the wearer is in a neutral/upright trunk posture, such as those employed by outdoor enthusiasts and schoolchildren. This research has led to some important harness design improvements that reduce trunk muscle exertions, fatigue and improve overall comfort. There are number of occupations, however, wherein workers wear back-mounted packs/devices (e.g. air tanks) while working in non-neutral trunk postures. The objective of the current study was to evaluate the effects of these non-neutral postures on biomechanical loading and then reconsider the backpack system design recommendations. Fifteen participants were asked to support a 18.2 kg load on their back while assuming static forward flexed postures of the torso (15 degrees , 30 degrees , 45 degrees , and 60 degrees of sagittal bend). The mass on the back was attached to the participant through two different harness mechanisms: a basic harness design (as seen on college student backpacks) and a more advanced design containing lateral stiffness rods and a weight-bearing hip belt (as seen on backpacks for hikers). While performing these static, posture maintenance tasks, the activation levels of the bilateral trapezius, erector spinae, and rectus abdominis were collected. Participants also provided subjective ratings of comfort. The results showed that there was a significant interaction between harness type and forward flexion angle for the trapezius and the erector spinae muscles. The normalized EMG for the trapezius muscles showed a 14% and 11% reduction in muscle activity at 15 degrees and 30 degrees , respectively, with the advanced design but these positive effects of the advanced design were not found at the greater flexion angles. Likewise the erector spinae muscles showed a 24% and 14% reduction in muscle activity at 15 degrees and 30 degrees , respectively, with the advanced design harness but these effects of the advanced design were not found at the greater forward flexion angles. The level of forward flexion angle affected the rectus abdominis muscle activity, but neither the harness type main effect nor the interaction of harness type and forward flexion angle was significant. The subjective survey results agreed with the EMG results and showed the advanced design harness was generally more comfortable with respect to the shoulder and low back areas. Collectively, the subjective and objective results show a significant improvement with the advanced harness system but also note an interesting interaction with degree of sagittal flexion, indicating a diminished effectiveness of the design improvements at forward flexed postures. Design criteria for harness systems in these forward flexed postures are discussed. PMID- 17112460 TI - Chewing chitosan-containing gum effectively inhibits the growth of cariogenic bacteria. AB - OBJECTIVE: We have already reported that chitosan inhibited the growth of cariogenic bacteria in vitro. This study was designed to evaluate whether chewing gum, containing chitosan, can effectively suppressed the growth of oral bacteria (total bacteria, total Streptococci, mutans streptococci (MS)) in saliva. METHODS: Fifty healthy subjects, ranging in age from 19 to 32 years, were recruited from among the staff and students of Nagasaki University School of Dentistry. For the slab of gum study, the subjects chewed gum for 5 min and then rested for 5 min. Each subject chewed a total of eight pieces of gum, which was either supplemented with or without chitosan, for a total of 80 min. Two different types of gums were examined with at least 1 week as a rest period in between treatments. This in vivo study was carried out by the double blind comparison test. RESULTS: The amount of oral bacteria was found to significantly decrease in the chitosan group. Especially, the number of MS were maintained at about a 20% level in comparison to that before gum chewing, even at 1h after gum chewing. CONCLUSION: These findings suggest that a supplementation of chitosan to gum is an effective method for controlling the number of cariogenic bacteria in situations where it is difficult to brush one's teeth, such as when an individual is away from home all day or participating in outdoor training. PMID- 17112461 TI - The automaticity of emotional Stroop: a meta-analysis. AB - An automatic bias to threat is often invoked to account for colour-naming interference in emotional Stroop. Recent findings by McKenna and Sharma [(2004). Reversing the emotional Stroop effect reveals that it is not what it seems: The role of fast and slow components. Journal of Experimental Psychology: Learning, Memory, and Cognition, 30, 382-392], however, cast doubt on the fast and non conscious nature of emotional Stroop. Interference by threat words only occurred with colour naming in the trial subsequent to the threat trial (i.e., a "slow" effect), but not immediately (i.e., a "fast" effect, as would be predicted by the bias hypothesis). In a meta-analysis of 70 published emotional Stroop studies the largest effects occurred when presentation of threat words was blocked, suggesting a strong contribution by slow interference. We did not find evidence; moreover, for interference in suboptimal (less conscious) presentation conditions and the only significant effects were observed in optimal (fully conscious) conditions with high-anxious non-clinical participants and patients. The emotional Stroop effect seems to rely more on a slow disengagement process than on a fast, automatic, bias. PMID- 17112462 TI - Implicit cognitive processes in psychopathology: an introduction. AB - Implicit or automatic processes are important in understanding the etiology and maintenance of psychopathological problems. In order to study implicit processes in psychopathology, measures are needed that are valid and reliable when applied to clinical problems. One of the main topics in this special issue concerns the development and validation of new or modified implicit tests in different domains of psychopathology. The other main topic concerns the prediction of clinical outcomes and new ways to directly influence implicit processes in psychopathology. We summarize the contributions to this special issue and discuss how they further our knowledge of implicit processes in psychopathology and how to measure them. PMID- 17112463 TI - Post-event processing in social anxiety. AB - Clark and Wells' [1995. A cognitive model of social phobia. In: R. Heimberg, M. Liebowitz, D.A. Hope, & F.R. Schneier (Eds.) Social phobia: Diagnosis, assessment and treatment (pp. 69-93). New York: Guildford Press.] cognitive model of social phobia proposes that following a social event, individuals with social phobia will engage in post-event processing, during which they conduct a detailed review of the event. This study investigated the relationship between self-appraisals of performance and post-event processing in individuals high and low in social anxiety. Participants appraised their performance immediately after a conversation with an unknown individual and prior to an anticipated second conversation task 1 week later. The frequency and valence of post-event processing during the week following the conversation was also assessed. The study also explored differences in the metacognitive processes of high and low socially anxious participants. The high socially anxious group experienced more anxiety, predicted worse performance, underestimated their actual performance, and engaged in more post-event processing than low socially anxious participants. The degree of negative post-event processing was linked to the extent of social anxiety and negative appraisals of performance, both immediately after the conversation task and 1 week later. Differences were also observed in some metacognitive processes. The results are discussed in relation to current theory and previous research. PMID- 17112464 TI - Supercritical carbon dioxide generated vascular endothelial growth factor encapsulated poly(DL-lactic acid) scaffolds induce angiogenesis in vitro. AB - The ability to deliver, over time, biologically active vascular endothelial growth factor-165 (VEGF) through tailored designed scaffolds offers tremendous therapeutic opportunities to tissue-engineered therapies. Porous biodegradable poly(DL-lactic) acid (PLA) scaffolds encapsulating VEGF have been generated using supercritical CO2 (scCO2) and the kinetic release and angiogenic activity of these scaffolds examined in vitro and in an ex vivo chick chorioallantoic membrane (CAM) angiogenesis model. After processing through scCO2, VEGF maintained its angiogenic activity as assessed by increased tubule formation of human umbilical vein endothelial cells (HUVEC) cultured on Matrigel (VEGF = 1937 +/- 205 microm; scCO2-VEGF = 2085 +/- 234 microm; control = 1237 +/- 179 microm). VEGF release kinetics from scCO2-VEGF incorporated PLA monolith scaffolds showed a cumulative release of VEGF (2837 +/- 761 rhog/ml) over a 21 day period in culture. In addition, VEGF encapsulated PLA scaffolds increased the blood vessel network in the CAM compared to controls; control, 24.8 +/- 9.6; VEGF/PLA, 44.1 +/ 12.1 (vessels/field). These studies demonstrate that the controlled release of growth factors encapsulated into three-dimensional PLA scaffolds can actively stimulate the rapid development of therapeutic neovascularisation to regenerate or engineer tissues. PMID- 17112465 TI - A novel embryotoxic estimation method of VPA using ES cells differentiation system. AB - Valproic acid (VPA), which has a wide range of therapeutic applications, is known as a potent teratogen that induces neural tube defects in vertebrates. Here, we have characterized the tissue-specific, embryotoxic effects of VPA on developmental processes using a novel system with differentiating mouse ES cells. Under our cultivating condition, ES cells differentiated into cardiomyocytes, although various cell types can be differentiated. VPA affected cell viability and differentiation from undifferentiated ES cells to cardiomyocytes in a dose dependent manner. The analysis of tissue-specific markers also revealed that VPA potently inhibited mesodermal and endodermal development but promoted neuronal differentiation in a lineage-specific manner. Taking the in vivo teratogenicity of VPA into account, this assay system could be useful in predicting the degree of embryotoxicity of VPA. We, thus, propose that the in vivo embryotoxic effects of various medicines can be estimated fast and accurately using this in vitro cell differentiation system. PMID- 17112466 TI - Translocation and interactions of L-arabinose in OmpF porin: A molecular dynamics study. AB - The passage of a natural substrate, L-arabinose (L-ARA) through Escherichia coli porin embedded in an artificial bilayer, is studied by equilibrium molecular dynamics simulations. We investigate the early stage of translocation process of L-ARA from intra-cellular to extra-cellular side (Int-to-Ext) across the bilayer. The average trajectory path over all L-ARA molecules along with quantum mechanical configuration-optimizations at PM3 level predict the existence of at least three trapping zones. The common feature within all these zones is that L ARA remains perpendicular to the channel axis. It is remarkable how the orientation and translational-rotational motion of L-ARA molecule play a role in its transport through OmpF channel. These simulations are important for better understanding of permeation process in OmpF channel. They also provide an insight into the chiral recognition of translocation process in protein nanochannels from substrate and protein prospects and help interpret experiments on permeation process of small dipolar molecules across biological membranes. PMID- 17112467 TI - High-resolution imaging of proteins in human teeth by scanning probe microscopy. AB - High-resolution studies of dental tissues are of considerable interest for biomedical engineering and clinical applications. In this paper, we demonstrate the application of piezoresponse force microscopy (PFM) to nanoscale imaging of internal structure of human teeth by monitoring the local mechanical response to an electrical bias applied via a conductive tip. It is shown that PFM is capable of detecting dissimilar components of dental tissues, namely, proteins and calcified matrix, which have resembling morphology but different piezoelectric properties. It is demonstrated that collagen fibrils revealed in chemically treated intertubular dentin exhibit high piezoelectric activity and can be visualized in PFM with spatial resolution of 10 nm. Evidence of the presence of protein inclusions of 100-200 nm wide and several micrometers long in tooth enamel has been obtained. Furthermore, it is found that the peritubular dentin and intertubular dentin exhibit different piezoelectric behavior suggesting different concentration of collagen fibrils. The obtained results demonstrate a high potential of PFM in providing an additional insight into the structure of dental tissues. It is suggested that the PFM approach can be used to study the structure of a wide range of biological materials by monitoring their electromechanical behavior at the nanoscale. PMID- 17112469 TI - Helper virus-independent trans-replication of hepatitis C virus-derived minigenome. AB - We have previously described a synthetic T7-driven cDNA minigenome containing the antisense sequence of luciferase gene and internal ribosome entry site of encephalomyocarditis virus flanked by 5'- and 3'-end sequences of hepatitis C virus (HCV) that contain cis-acting replication elements. Synthesis of minus strand RNA from the artificial minigenome was determined by using Huh-7 cells harboring autonomously replicating HCV subgenome as a helper for provision of functional replication components. To further confirm and extend these studies, we investigated here whether the minigenome replication system could be reconstituted by transfection of naive Huh-7 cells with plasmid expressing nonstructural (NS) proteins. Reporter assay and Northern blot analysis revealed that trans-expression of NS proteins from 3 to 5 resulted in high level of luciferase activity and synthesized minus-strand RNA. The analogous result was also obtained with the minigenome derived from HCV 2a, and both HCV 1b- and 2a derived NS protein were able to support the chimeric minigenomes whose 5'- or 3' end was replaced by the respective region of the heterologous virus. These results provide a basis for establishing the reverse genetic system that is helpful to study cis- and trans-acting factors involved in HCV RNA replication. PMID- 17112468 TI - Dose-dependent inhibitory effect of CD47 in macrophage uptake of IgG-opsonized murine erythrocytes. AB - The cell surface glycoprotein CD47 on target cells can bind to the inhibitory receptor SIRPalpha on macrophages to inhibit phagocytosis of antibody sensitized blood cells. The aim of this study was to determine if CD47 dose-dependently can regulate macrophage uptake of IgG-opsonized RBCs. CD47(+/-) RBCs express about 50% of the CD47 level found on CD47(+/+) RBCs. When injected into CD47(+/+) mice, CD47(+/-) RBCs showed a significantly faster antibody-mediated clearance as compared with CD47(+/+) RBCs injected into the same recipient. In vitro phagocytosis experiments confirmed that CD47(+/-) RBCs were taken up significantly more than CD47(+/+) RBCs, but significantly less than CD47(-/-) RBCs. A reduction in RBC CD47 expression just below 50% of that in normal RBCs can significantly accelerate RBC clearance by macrophages in the presence of RBC autoantibodies. This may have relevance for transfusion of stored RBCs, where loss of CD47 is seen over time, and in clearance of these cells by antibody dependent phagocytosis. PMID- 17112470 TI - Progressive mRNA decay establishes an mkp3 expression gradient in the chick limb bud. AB - The apical ectodermal ridge (AER) controls limb outgrowth and patterning, such that its removal causes changes in mesodermal gene expression, cell death and limb truncation. Fibroblast growth factor (FGF) family members are expressed in the AER and can rescue limb bud outgrowth after AER removal. Cells localized underneath the AER are maintained in an undifferentiated state by the FGFs produced by the AER. MAPK phosphatase 3 (mkp3) is a downstream effector of FGF8 signalling during limb bud development and is expressed in the distal limb mesenchyme. The present work evidences a gradient of mkp3 transcripts along the chick limb bud, in a distal to proximal direction. mkp3 transcription occurs only in the most distal limb bud cells and its mRNA gradient throughout the limb results from progressive mRNA decay. We show that FGF8-soaked beads induce ectopic mkp3 expression, indicating that AER-derived FGF8 protein may activate mkp3 in the distal mesenchyme. PMID- 17112471 TI - ADAM19 is tightly associated with constitutive Alzheimer's disease APP alpha secretase in A172 cells. AB - To elucidate whether new proteases are involved in the processing of amyloid precursor protein (APP), we examined catalytically active ADAM12 and ADAM19 as candidates alpha-secretases. The overexpression of ADAM19 in HEK293 cells resulted in an increase in sAPPalpha. Therefore, we suggest that ADAM19 has a constitutive alpha-secretase activity. We examined regulated alpha-secretase activity by adding phorbol 12-myristate 13-acetate (PMA), but no regulated activity was found. To verify that endogenous ADAM19 has an APP alpha-secretase activity, we examined whether the constitutive level of alpha-secretase activity was reduced by RNA interference with ADAM19 in A172 cells. The amount of secreted sAPPalpha decreased by about 21% following RNAi. These results suggest that ADAM19 has a constitutive alpha-secretase activity for APP. PMID- 17112472 TI - Site-directed mutagenesis of conserved aromatic residues in rat squalene epoxidase. AB - Squalene epoxidase catalyzes the conversion of squalene to (3S)2,3-oxidosqualene, which is a rate-limiting step of the cholesterol biogenesis. To evaluate the importance of conserved aromatic residues, 15 alanine-substituted mutants were constructed and tested for the enzyme activity. Except F203A, all the mutants significantly lost the enzyme activity, confirming the importance of the residues, either for correct folding of the protein, or for the catalytic machinery of the enzyme. Further, interestingly, F223A mutant no longer accepted (3S)2,3-oxidosqualene as a substrate, while Y473A mutant converted (3S)2,3 oxidosqualene to (3S,22S)2,3:22,23-dioxidosqualene twice more efficiently than wild-type enzyme. It is remarkable that the single amino acid replacement yielded mutants with altered substrate and product specificities. These aromatic residues are likely to be located at the substrate-binding domain of the active-site, and control the stereochemical course of the enzyme reaction. PMID- 17112473 TI - Opposite effects of endotoxin on mitochondrial and endoplasmic reticulum functions. AB - In this study, we determined functional integrity and reactive oxygen species generation in mitochondria and endoplasmic reticulum in liver of rats subjected to endotoxic shock to clarify whether intracellular reactive oxygen species (ROS) destabilize cellular integrity causing necrosis in rats challenged with lipopolysaccharide (LPS). LPS caused drastically increased plasma levels of alanine aminotransferase, suggesting damage to plasma membranes of liver cells. Liver necrosis was confirmed by histological examination. LPS induced a significant increase in ROS production in rat liver mitochondria (RLM), but did not impair mitochondrial function. In contrast to mitochondria, enzymatic activity and ROS production of cytochrome P450 were lower in microsomal fraction obtained from LPS-treated animals, suggesting the dysfunction of endoplasmic reticulum. Protein patterns obtained from RLM by two-dimensional electrophoresis showed significant upregulation of mitochondrial superoxide dismutase by LPS. We hypothesize that upregulation of this enzyme protects mitochondria against mitochondrial ROS, but does not protect other cellular compartments such as endoplasmic reticulum and plasma membrane causing necrosis. PMID- 17112474 TI - Molecular cloning and characterization of mouse aquaporin 6. AB - In the rat kidney, aquaporin (AQP) 6 is localized in the intracellular vesicle membranes of type-A intercalated cells of the collecting duct; mouse AQP6 (mAQP6) has not been characterized. Although mAQP6 was originally cloned from cDNA in a mouse cerebellum library (GenBank NM 175087), we have independently cloned a cDNA encoding mAQP6 from an adult kidney cDNA library (C57BL/6J strain). We identified two different spliced variants of mAQP6: mAQP6a and mAQP6b. The mAQP6a isoform is almost identical to that of rat AQP6, whereas mAQP6b is identical to that reported in the mouse cerebellum library mentioned above. We found that the mRNA expression of these two spliced variants is regulated in a tissue-specific and age-dependent manner. Functional analyses of water and ion permeation revealed that mAQP6a functions like rat AQP6 and that mAQP6b does not function as either a water channel or an ion channel under our experimental conditions. PMID- 17112475 TI - The zebrafish genome contains two inducible, functional cyclooxygenase-2 genes. AB - Cyclooxygenase is a key enzyme in prostanoid biosynthesis. Mammalian species have two cyclooxygenases, constitutively expressed cyclooxygenase-1 (Cox-1) and inducible cyclooxygenase-2 (Cox-2). Cox-1 and/or Cox-2 have been also identified in other vertebrates, including fish. We identified a second zebrafish Cox-2 gene orthologue, Cox-2b. All of the functionally important amino acids for cyclooxygenase enzymes are conserved in Cox-2b. The 3' untranslated region of the Cox-2b message contains AU rich elements characteristic of regulation at the level of mRNA stability. Constitutive tissue expression patterns for Cox-2a and Cox-2b are distinct, but overlap. Both Cox-2a and Cox-2b expression are inducible in the kidney when fish are exposed to tetradecanoylphorbol acetate. Like Cox-2a, Cox-2b protein, expressed in COS cells is functionally active. Thus, the zebrafish genome contains two functional, inducible Cox-2 genes. Database searching demonstrates that some fish genomes contain multiple Cox-1 or Cox-2 cyclooxygenase genes, suggesting alternate duplication and retention of this gene. PMID- 17112476 TI - Can chicken and human PrPs possess SOD-like activity after beta-cleavage? AB - The prion protein is a membrane attached glycoprotein that is involved in binding of divalent copper ions. In vivo human and chicken PrPs exhibit SOD-like activity associated with octarepeat and hexarepeat regions, respectively, when bind Cu(II) ions. However, the species of Cu(II)-PrP involved in the Cu(II) center which determines the highest SOD-like activity is still unknown. The data presented here clearly show that the single Cu(II) ion bound to PrP octapeptide repeat region of mammalian prion and hexapeptide repeat region of avian prion via 4 His side-chain imidazoles reveals the highest SOD activity. PMID- 17112477 TI - Functional characterization of a mammalian transcription factor, Elongin A. AB - Elongin A is the transcriptionally active subunit of the Elongin complex that strongly stimulates the rate of elongation by RNA polymerase II (pol II) by suppressing the transient pausing of the polymerase at many sites along the DNA template. We have recently shown that Elongin A-deficient mice are embryonic lethal, and mouse embryonic fibroblasts (MEFs) derived from Elongin A(-/-) embryos display not only increased apoptosis but also senescence-like phenotypes accompanied by the activation of p53. To further understand the function of Elongin A in vivo, we have carried out the structure-function analysis of Elongin A and identified sequences critical to its nuclear localization and direct interaction with pol II. Moreover, we have analyzed the replication fork movement in wild-type and Elongin A(-/-) MEFs, and shown the possibility that the genomic instability observed in Elongin A(-/-) MEFs might be caused by the replication fork collapse due to Elongin A deficiency. PMID- 17112478 TI - Na+ entry via TRPC6 causes Ca2+ entry via NCX reversal in ATP stimulated smooth muscle cells. AB - Reversal of the Na+/Ca2+ -exchanger (NCX) has been shown to mediate Ca2+ influx during activation of G-protein linked receptors. Functional coupling between the reverse-mode NCX and the canonical transient receptor potential channels (TRPCs) has been proposed to mediate Ca2+ influx in HEK-293 cells overexpressing TRPC3. In this communication we present evidence for similar functional coupling of NCX to endogenously expressed TRPC6 in rat aorta smooth muscle cells. Selective inhibition of reverse-mode NCX with KB-R7943 and of non-selective cation-channels with SKF-96365 abolished Ca2+ influx in response to agonist stimulation (ATP). Expression of a dominant negative TRPC6 mutant also reduced the Ca2+ influx in proportion to its transfection efficiency. Calyculin A, which is known to disrupt the junctions of the plasma membrane and sarco/endoplasmic reticulum, increased global Na+ elevations and reduced stimulated Ca2+ influx. Together our data provide evidence that localized Na+ elevations are generated by TRPC6 and drive reversal of NCX to mediate Ca2+ influx. PMID- 17112479 TI - Effect of drug-induced ascorbic acid release in the striatum and the nucleus accumbens in hippocampus-lesioned rats. AB - The mechanism of ethanol, morphine, methamphetamine (MAP), and nicotine-induced ascorbic acid (AA) release in striatum, and nucleus accumbens (NAc) is not well understood. Our previous study showed that the glutamatergic system was involved in the addictive drug-induced AA release in NAc and striatum. Furthermore, frontal decortication eliminates drug-induced ascorbic acid release in the striatum but not in the NAc. In the present study, the roles of the hippocampus in drug-induced AA release in the striatum and NAc were studied by using microdialysis coupled to high performance liquid chromatography with electrochemical detection (HPLC-ECD). Ethanol (3.0 g/kg, i.p.), methamphetamine (3.0 mg/kg, i.p.), and nicotine (1.5 mg/kg, i.p.) significantly stimulated AA release in the striatum and NAc, respectively. Morphine (20 mg/kg, i.p.) significantly stimulated AA release in the striatum, but not in the NAc. After hippocampal lesion by kainic acid, AA release induced by ethanol, methamphetamine, and nicotine could be eliminated in NAc, but not in the striatum. These results suggest that the hippocampus might be a common and necessary area in addictive drug-induced AA release in the NAc, which also imply that different pathways might be involved in drug-induced AA release in the striatum and the NAc of the rats. PMID- 17112480 TI - Myelination and nodal formation of regenerated peripheral nerve fibers following transplantation of acutely prepared olfactory ensheathing cells. AB - Transplantation of olfactory ensheathing cells (OECs) into injured spinal cord results in improved functional outcome. Mechanisms suggested to account for this functional improvement include axonal regeneration, remyelination and neuroprotection. OECs transplanted into transected peripheral nerve have been shown to modify peripheral axonal regeneration and functional outcome. However, little is known of the detailed integration of OECs at the transplantation site in peripheral nerve. To address this issue, cell populations enriched in OECs were isolated from the olfactory bulbs of adult green fluorescent protein (GFP) expressing transgenic rats and transplanted into a sciatic nerve crush lesion which transects all axons. Five weeks to 6 months after transplantation, the nerves were studied histologically. GFP-expressing OECs survived in the lesion and distributed longitudinally across the lesion zone. The internodal regions of individual teased fibers distal to the transection site were characterized by GFP expression in the cytoplasmic and nuclear compartments of cells surrounding the axons. Immunoelectron microscopy for GFP indicated that the transplanted OECs formed peripheral type myelin. Immunostaining for sodium channel and Caspr revealed a high density of Na(v)1.6 at the newly formed nodes of Ranvier which were flanked by paranodal Caspr staining. These results indicate that transplanted OECs extensively integrate into transected peripheral nerve and form myelin on regenerated peripheral nerve fibers, and that nodes of Ranvier of these axons display proper sodium channel organization. PMID- 17112481 TI - Binocular interactions in the guinea pig's visual-evoked potentials. AB - In this study, binocular interaction in guinea pigs is evaluated using bioelectrical activities. A difference potential, as evidence of an interaction, is calculated by subtracting the sum of visual-evoked potentials recorded by left and right monocular visual stimulations from the potential recorded by binocular stimulation. A negative monophasic wave with an average amplitude of 15.1 microV and an average latency of 106 ms is observed in the difference potential. This finding implies that the P100 is the main guinea pig visual-evoked potential wave that is affected by binocular interaction. Binocular interaction is also observed in the waves N75 and N140, although with a smaller amplitude. No interaction is observed in the segments of P55 and P200 waves. PMID- 17112482 TI - Green love is ugly: emotions elicited by synesthetic grapheme-color perceptions. AB - Synesthetes who experience grapheme-color synesthesia often report feeling uneasy when dealing with incongruently colored graphemes although no empirical data is available to confirm this phenomenon. We studied this affective reaction related to synesthetic perceptions by means of an evaluation task. We found that the perception of an incorrectly colored word affects the judgments of emotional valence. Furthermore, this effect competed with the word's emotional valence in a categorization task thus supporting the automatic nature of this synesthetically elicited affective reaction. When manipulating word valence and word color photism congruence, we found that responses were slower (and less accurate) for inconsistent conditions than for consistent conditions. Inconsistent conditions were defined as those where semantics and color-photism congruence did not produce a similar assessment and therefore gave rise to a negative affective reaction (i.e., positive-valence words presented in a color different from the synesthete's photism or negative-valence words presented in the photism's color). We therefore observed a modulation of the congruency effect (i.e., faster reaction times to congruently colored words than incongruently colored words). Although this congruence effect has been taken as an index of the true experience of synesthesia, we observed that it can be reversed when the experimental manipulations turn an incongruently colored word into a consistent stimulus. To our knowledge, this is the first report of an affective reaction elicited by the congruency between the synesthetically induced color of a word and the color in which the word is actually presented. The underlying neural mechanisms that might be involved in this phenomenon are discussed. PMID- 17112483 TI - Effects of Raf-1 siRNA on human cerebral microvascular endothelial cells: a potential therapeutic strategy for inhibition of tumor angiogenesis. AB - The serine/threonine kinase Raf-1 is involved in the regulation of tumor cell survival, proliferation and metastasis formation, and has therefore emerged as a promising target for cancer therapy. In addition, Raf-1 activity mediates proliferation of endothelial cells thereby promoting angiogenesis and invasive growth of various tumors, including highly vascularized malignant glioblastoma. The aim of this study was to evaluate the effects of small inhibitory RNA (siRNA) directed against Raf-1 on viability, proliferation and motility in glioma cells and cerebral endothelial cells. Half-quantitative RT-PCR and Western blotting revealed efficient siRNA-mediated Raf-1 down regulation in glioma cells (U373, U251) and in human cerebral microvascular endothelial cells (HCMEC). Surprisingly, Raf-1 gene silencing failed to affect cell survival, proliferation or migration activity in the glioblastoma cell lines. In HCMEC, however, pronounced decrease of cell survival and significant inhibition of tube formation was achieved by Raf-1 siRNA compared to non-functional siRNA or vehicle controls. In conclusion, Raf-1 silencing appears as a potential therapeutic strategy to inhibit brain tumor angiogenesis and thereby outgrowth of highly vascularized glioblastoma multiforme, whereas direct cytotoxic effects of Raf-1 knockdown in tumor cells may vary. PMID- 17112484 TI - RVLM glycine receptors mediate GABAA and GABAB)independent sympathoinhibition from CVLM in rats. AB - The caudal ventrolateral medulla (CVLM) provides tonic inhibitory and also excitatory inputs to the rostral ventrolateral medulla (RVLM). These experiments evaluated the role of RVLM gamma-amino butyric acid (GABA) receptor subtypes and glycine receptors in mediating CVLM sympathoinhibition. In Inactin anesthetized female rats, the CVLM and RVLM were functionally defined by pressor and depressor responses to microinjected GABA (500 pmol, 50 nl). Although reduced, pressor and sympathoexcitatory responses due to inhibition of the CVLM with GABA persisted following ipsilateral RVLM GABA(A) receptor blockade (bicuculline, BIC, 400 pmol, 100 nl; n=12) in rats with contralateral nucleus tractus solitarius (NTS) lesion. In the presence of either ipsilateral (+contralateral NTS lesion; n=8) or bilateral (n=6) GABA(A) and GABA(B) receptor blockade of the RVLM (400 pmol BIC+400 pmol CGP35348, 100 nl), inhibition of the CVLM still increased MAP and renal sympathetic nerve activity (RSNA). Thus neither GABA(B) receptors nor a contralateral CVLM to RVLM GABAergic pathway explains residual responses to CVLM blockade. The addition of strychnine (300 pmol, 100 nl) to the RVLM eliminated responses to CVLM inhibition, suggesting that a GABA(A) and GABA(B) independent sympathoinhibitory influence from CVLM to RVLM is mediated by glycine receptors. Decreases in MAP and RSNA due to activation of the CVLM with glutamate (500 pmol, 50 nl) were reversed to increases in the presence of RVLM GABA(A) receptor blockade (n=7). Thus, a sympathoexcitatory pathway from the CVLM can be activated in the presence of RVLM GABA receptor blockade, but sympathoinhibitory influences from the CVLM predominate. PMID- 17112486 TI - Lexical mediation and context effects in sentence processing. AB - Studies of syntactic ambiguity resolution have played a central role in resolving questions about when and how contextual information affects parsing processes. These investigations are often couched in terms of modularity versus interaction, with demonstrations of rapid contextual effects being taken as evidence that the mechanisms responsible for structuring sentences are permeable to referential or semantic context, and therefore non-modular. In this paper, we will propose that argument relations are constructed on the basis of lexically stored syntactic representations (as in MacDonald, M.C., Pearlmutter, N.J., and Seidenberg, M.S. (1994). Lexical nature of syntactic ambiguity resolution. Psychological Review, 101, 676-703. Pickering, M.J., and Traxler, M.J. (2004). Grammatical repetition and garden path effects. Paper presented to the CUNY Sentence Processing Conference. College Park, MD., Pickering, M.J., and Traxler, M.J. (2006). Syntactic Priming in Comprehension. Manuscript in preparation. Traxler, M.J., and Pickering, M.J. (2005, March). Syntactic priming in comprehension. Paper presented to the CUNY Sentence Processing Conference. Tucson, AZ), but that other types of structural decisions are made on the basis of general processing principles. This formulation can be tested by looking at how the parser reacts to immediate intra- and inter-sentential factors (short-term context) and how it reacts to patterns of input over longer time scales (long-term context). We begin with a brief review of work on context effects in syntactic disambiguation, sketch our account of parsing, and then provide evidence from two eye-tracking experiments that illustrate some of the processing principles that govern parsing of argument relations. PMID- 17112485 TI - PKU is a reversible neurodegenerative process within the nigrostriatum that begins as early as 4 weeks of age in Pah(enu2) mice. AB - Phenylketonuria (PKU) is a common genetic disorder in humans that arises from deficient activity of phenylalanine hydroxylase (PAH), which catalyzes the conversion of phenylalanine to tyrosine. There is a resultant hyperphenylalanemia with subsequent impairment in cognitive abilities, executive functions and motor coordination. The neuropathogenesis of the disease has not been completely elucidated, however, oxidative stress is considered to be a key feature of the disease process. Hyperphenylalanemia also adversely affects monoaminergic metabolism in the brain. For this reason we chose to evaluate the nigrostriatum of Pah(enu2) mice, to determine if alterations of monoamine metabolism resulted in morphologic nigrostriatal pathology. Furthermore, we believe that recent developments in adeno-associated virus (AAV)-based vectors have greatly increased the potential for long-term gene therapy and may be a viable alternative to dietary treatment for this metabolic disorder. In this study we identified neurodegenerative changes with regenerative responses in the nigrostriatum of Pah(enu2) mice that are consistent with oxidative injury and occurred as early as 4 weeks of age. These neuropathologic changes were reversed following portal vein delivery of a recombinant adeno-associated virus-mouse phenylalanine hydroxylase woodchuck hepatitis virus post-transcriptional response element (rAAV-mPAH-WPRE) vector to Pah(enu2) mice and corresponded to rapid reduction of serum Phe levels. PMID- 17112487 TI - Postnatal development of dopamine innervation in the amygdala and the entorhinal cortex of the gerbil (Meriones unguiculatus). AB - Dopamine (DA) projections from the mesencephalon are believed to play a critical role during development and are essential for cognitive and behavioral functions. Since the postnatal maturation patterns of these projections differ substantially between various brain regions, cortical, limbic or subcortical areas might exhibit varying vulnerabilities concerning developmental disorders. The dopaminergic afferents of the rodent prefrontal cortex show an extremely prolonged maturation which is very sensitive to epigenetic challenges. However, less is known about the development of the DA innervation of caudal limbic areas. Therefore, immunohistochemically stained DA fibers were quantitatively examined in the basolateral (BLA) and central amygdaloid nucleus (CE) and the ventrolateral entorhinal cortex (EC) of the Mongolian gerbil (Meriones unguiculatus). Animals of different ages, ranging from juvenile [postnatal day (PD) 14, 20, 30)] to adolescent (PD70), adult (6, 18 months) and aged (24 months), were analyzed. Results show a significant increase of fibers between PD14 and PD20 in the BLA and lateral part of the CE, with a trend for a subsequent decline in fiber densities until PD30. The EC and medial part of the CE showed no developmental changes. Interestingly, none of the investigated areas showed significant reductions of DA fibers during aging. PMID- 17112488 TI - Neprilysin protects human neuronal progenitor cells against impaired development caused by amyloid-beta peptide. AB - Transplantation of human neuronal progenitor cells (HNPC) is being considered for neuroreplacement therapy in beta-amyloidosis associated with neuronal loss in Down's syndrome and Alzheimer's disease. However, the influence of amyloid-beta containing brain environment on the development of HNPCs is unknown. Recently, we demonstrated that amyloid-beta peptide (Abeta) impaired differentiation of HNPCs in culture through oxidative stress. Now we studied the effect of neprilysin, an Abeta-degrading enzyme, on development of neuronal colonies from neurospheres of HNPCs in the presence of Abeta1-40. Neprilysin increased the number of neurospheres that formed colonies of neuron-like cells. This effect of neprilysin was associated with reduced amounts of the monomeric and dimeric Abeta that remained in culture supernatants as well as the Abeta uptaken by differentiating HNPCs. Phosphoramidon, a neprilysin inhibitor, attenuated these effects of neprilysin. In control cultures of HNPCs that grew without exogenous Abeta1-40, the treatment with neprilysin reduced the number of developing colonies. This effect might result from degradation by neprilysin of endogenous Abeta produced and secreted by HNPCs or other peptides that are involved in neuronal development. The results demonstrate that even a partial reduction of extracellular Abeta levels by neprilysin may facilitate development of HNPCs into neurons in an environment overloaded with Abeta. This finding suggests that neprilysin could facilitate neuroreplacement therapy with HNPCs in treatment of neurodegenerative diseases. PMID- 17112489 TI - One-pot synthesis of 2-C-glycosylated benzimidazoles from the corresponding methanal dimethyl acetals. AB - A series of 2-glycosyl-benzimidazoles with alpha-d-arabinopyranosyl, beta-d galactopyranosyl, beta-d-glucopyranosyl, beta-d-mannopyranosyl, and beta-l rhamnopyranosyl configurations were obtained in 52-73% yields from the corresponding C-glycosylmethanal dimethyl acetals and o-phenylenediamine under catalysis with hydrogen chloride or a strongly acidic cation-exchange resin. Intermediate benzimidazolines were spontaneously oxidized by air to produce the final products in the one-pot procedure. The prepared compounds did not show any inhibitory effect on the growth of 12 strains of five different species of pathogenic yeasts. PMID- 17112490 TI - Synthesis and evaluation of glycosyl donors with novel leaving groups for transglycosylations employing beta-galactosidase from bovine testes. AB - Novel aryl beta-d-galactopyranosides were synthesized employing phase-transfer catalysis, and assayed as potential galactose donors in the presence of beta galactosidase from bovine testes using pNP-Gal as a reference. The aglycones were represented mainly by nitrophenols containing halogens, hydroxymethyl, aldehyde, carboxyl, ester or amino functions. An unusual intermolecular acetyl migration onto the benzylic alcohol group was observed during galactosylation of hydroxymethylnitrophenols. Pyridyl glycosides were obtained by reaction with the corresponding silver pyridinolates. Glycosides of halo-, hydroxymethyl- or methoxycarbonyl-nitrophenols as leaving groups gave virtually the same yields of transglycosylation products. A minor increase was achieved with nitrosalicylaldehyde as leaving group, whereas carboxy or amino derivatives gave very low or no yield of the transglycosylation product. Commercially available donors such as resorufinyl and 4-methylumbelliferyl beta-d-galactopyranosides exhibited a lower transglycosylation potential than these novel pNP-Gal derivatives. PMID- 17112491 TI - Determination of the maximum water solubility of eight native starches and the solubility of their acidic-methanol and -ethanol modified analogues. AB - The maximum water solubilities of eight native starches from potato, shoti, tapioca, maize, waxy maize, amylomaize-7, wheat, and rice and their acid-methanol and acid-ethanol modified analogues have been determined. Maximum solubilities of 18.7 and 17.4 mg/mL were obtained for waxy maize and tapioca and 12.4 mg/mL for potato and maize starches by autoclaving 220 mg/10 mL at 121 degrees C; 8.7 mg/mL was obtained for shoti starch by stirring in 85:15 (v/v) Me(2)SO-H(2)O at 20 degrees C; and 7.0 and 5.2mg/mL for rice and amylomaize-7 starches by stirring in 1M NaOH at 20 degrees C. The acid-alcohol treated starches were 4-9 times more soluble than their native starches. The compositions of the solubilized starches had, in general, much higher ratios of amylose to amylopectin than the ratios in their native granules. A major exception to this was the acid-methanol treated potato, shoti, and rice starches that had much lower ratios of amylose to amylopectin than the ratios in their granules. PMID- 17112492 TI - Cholecalciferol treatment changes urinary sodium-potassium ratio and plasma aldosterone of spontaneously hypertensive rats. PMID- 17112493 TI - Determination of kynurenic acid in human serum and its correlation with the concentration of certain amino acids. AB - BACKGROUND: Kynurenic acid (KYNA)--a tryptophan metabolite--elicits antagonistic activity against glutaminergic and cholinergic receptors; it has been suggested to have some relationship with neurological disorders. Considering this, serum KYNA may be an important marker in clinical diagnosis. We determined serum KYNA concentration and elucidate its correlation with several amino acids in human serum. METHODS: KYNA and amino acids concentrations in human serum of healthy subjects [n=35 (21 males and 14 females)] were determined by HPLC with fluorescence detection; thus, the correlation between KYNA concentration and that of several amino acids was examined in these subjects. RESULTS: Of the amino acids examined in this study, a significant negative correlation was observed between KYNA and glutamine (Gln) concentrations (r=-0.452, p<0.01) in the healthy subjects, particularly males (r=-0.687, p<0.01), and age-related changes were not observed. In addition to Gln, Gly and Ala concentrations showed a significant negative correlation with KYNA concentration in the serum of male subjects (r= 0.440 and -0.456, respectively, p<0.05). CONCLUSION: The significant correlation between KYNA and Gln concentrations in vivo may support the previous finding that kynurenine aminotransferase I (KAT I), responsible for the biosynthesis of KYNA, was identical to Gln transaminase K (GTK), which catalyses the transamination of Gln to 2-oxoglutamic acid. Both KYNA and Gln concentrations in vivo might be influenced due to altered KAT I/GTK activity. PMID- 17112494 TI - Association of adiponectin and amino terminal proBNP in peripheral arterial disease. AB - BACKGROUND: The aim of the present study was to investigate the relationship of adiponectin, a novel adipocytokine, and amino terminal proBNP (NT-proBNP) in patients with peripheral arterial disease (PAD). METHODS: Serum concentrations of adiponectin and NT-proBNP were measured in 487 patients with symptomatic PAD from the Linz Peripheral Arterial Disease (LIPAD) study. RESULTS: Correlation analysis revealed an association of adiponectin and NT-proBNP (r, +0.47; p<0.001). Even after adjustment for age, sex, body mass index, diabetes mellitus, smoking, arterial hypertension, estimated glomerular filtration rate (eGFR), fasting glucose, LDL-cholesterol, HDL-cholesterol, triglycerides, high-sensitivity C reactive protein, and total homocysteine the relationship of adiponectin and NT proBNP remained significant (r, +0.35; p<0.001). Furthermore, a subgroup analysis of patients with first manifestation of symptomatic PAD (n=287) demonstrated that disease severity (classified by Fontaine stages) was positively related to adiponectin (r, +0.13; p=0.003) and NT-proBNP (r, +0.28; p<0.001). CONCLUSION: Adiponectin was positively associated with NT-proBNP in symptomatic atherosclerotic PAD, independent of traditional and non-traditional risk factors. Moreover, adiponectin and NT-proBNP were related to disease severity, indicating a possible role for assessment of future morbidity and mortality in patients with PAD. PMID- 17112495 TI - Determination of glucuronidated 5-hydroxytryptophol (GTOL), a marker of recent alcohol intake, by ELISA technique. AB - OBJECTIVES: To compare markers of alcohol consumption. DESIGN AND METHODS: Measurement of urinary ethyl glucuronide, 5-hydroxytryptophol, 5 hydroxytryptophol glucuronide, 5-hydroxyindole acetic acid in 10 patients during alcohol withdrawal. RESULTS: 5-Hydroxytryptophol glucuronide was measured by ELISA with good analytical precision, its diagnostic specificity and sensitivity was better than that of 5-hydroxytryptophol and its correlation was closer to ethyl glucuronide than to 5-hydroxytryptophol. CONCLUSION: Determination of 5 hydroxytryptophol glucuronide by ELISA offers promising results in detection of previous alcohol consumption. PMID- 17112496 TI - Stone detection in MRCP images using controlled region growing. AB - Stones in the biliary tract are routinely identified using MRCP (magnetic resonance cholangiopancreatography). The noisy nature of the images, as well as varying intensity, size and location of the stones, defeat most automatic detection algorithms, making computer-aided diagnosis difficult. This paper proposes a multi-stage segment-based scheme for semi-automated detection of choledocholithiasis and cholelithiasis in the MRCP images, producing good performance in tests, differentiating them from "normal" MRCP images. With the high success rate of over 90%, refinement of the scheme could be applicable in the clinical environment as a tool in aiding diagnosis, with possible applications in telemedicine. PMID- 17112497 TI - A smart virtual glove for the hand telerehabilitation. AB - Hand rehabilitation, following stroke or hand surgery, is repetitive and long duration and can be facilitated with the assistance of complex, heavy and cumbersome haptic gloves based on sensors. The present paper describes a virtual glove, software based, which tracks hand movements by using images collected from webcams and numerical analysis. Finger forces are calculated from the deformations impressed to some objects (of known elastic coefficient) grasped by the patient hand. The presented system is notable for simplicity, generality and low cost. Implementation and results of the proposed virtual glove will be the objects of a future paper. PMID- 17112498 TI - The control of growth and differentiation of the wing imaginal disks of Manduca sexta. AB - During the larval feeding period, the growth of the wing imaginal disks of Lepidoptera is dependent on continuous feeding. Feeding and nutrition exert their effect via the secretion of bombyxin, the lepidopteran insulin-like hormone. When larvae stop feeding and enter the wandering stage in preparation for metamorphosis, the control of imaginal disk growth becomes feeding and nutrition independent. Growth of the wing imaginal disks of non-feeding wandering stage Manduca sexta can be stopped by removal of the brain, indicating that a brain derived factor is required for continued disk growth. Isolated wing disk growth in vitro requires both 20-hydroxyecdysone (20E) and either brain extract or bombyxin to achieve normal growth. In vitro, brain extracts and synthetic bombyxin have little or no effect in stimulating disk growth, but they greatly enhance the effect of 20E, indicating that 20E and bombyxin act synergistically to modulate growth of the imaginal disk. Brain extract and bombyxin induce a suite of insulin-response events in cultured wing disks, which indicate that bombyxin and 20E act through separate and synergistic pathways. The dose-response to 20E reaches a plateau at about 0.1 microg/ml. Tracheal differentiation of the wing disks can be induced to initiate in vitro by a low concentration of 20E, whereas higher concentrations of 20E only stimulate growth. PMID- 17112499 TI - Mucosal acid causes gastric mucosal microcirculatory disturbance in nonsteroidal anti-inflammatory drug-treated rats. AB - The mechanism by which nonsteroidal anti-inflammatory drugs (NSAIDs) suppress gastric mucosal blood flow is not fully understood, although the depletion of mucosal prostaglandin E2 has been proposed as one possible explanation. We investigated the role of gastric acid on gastric mucosal blood flow in NSAID treated rats. A rat stomach was mounted in an ex vivo chamber, and gastric mucosal blood flow was measured sequentially in a 5-mm2 area of the gastric corpus using a scanning laser Doppler perfusion image system. Results showed that diclofenac (5 mg/kg s.c.) and indomethacin (10 mg/kg s.c.) did not affect gastric mucosal blood flow, although both strongly decreased mucosal prostaglandin E2 when saline was instilled into the gastric chamber. On replacement of the saline in the chamber with 100 mM hydrochloric acid, these drugs caused a decrease in gastric mucosal blood flow levels within 30 min. The specific cyclooxygenase (COX)-2 inhibitors celecoxib (50 mg/kg s.c.) and rofecoxib (25 mg/kg s.c.) did not affect mucosal prostaglandin E2 level, nor did they decrease gastric mucosal blood flow, even when hydrochloric acid was added to the chamber. Furthermore, measurement of vasoconstrictive factors present in the mucosa showed that endothelin-1 levels increased after administration of diclofenac s.c. in the presence of intragastric hydrochloric acid. This indicates that the presence of mucosal hydrochloric acid plays an important role in the NSAID-induced decrease in gastric mucosal blood flow, while the COX-1-derived basal prostaglandin E2, which is unlikely to control gastric mucosal blood flow itself, protects microcirculatory systems from mucosal hydrochloric acid. PMID- 17112500 TI - Interactive effect of histamine and prostaglandin D2 on nasal allergic symptoms in rats. AB - This study was undertaken to investigate the interactive effect of histamine and prostaglandin D(2) in nasal allergic symptoms in rats. The intranasal application of histamine at doses lower than 10 mumol/site caused no sneezing or nasal rubbing. In addition, prostaglandin D(2) also showed no significant increase in these responses, even at a dose of 10 nmol/site. On the other hand, the simultaneous instillation of histamine and prostaglandin D(2) resulted in a 1000 times more potent effect in inducing nasal symptoms than the administration of histamine alone. Thus, prostaglandin D(2) enhanced the actions of histamine in inducing sneezing and nasal rubbing in a dose-dependent manner, and significant effects were observed at doses higher than 1 nmol/site. The responses induced by the simultaneous application of histamine and prostaglandin D(2) were inhibited by chlorpheniramine, cyproheptadine, BW A868C and ramatroban. Chlorpheniramine and cyproheptadine showed the dose-related inhibition of nasal symptoms induced by the combined administration of histamine (10 nmol) and prostaglandin D(2) (10 nmol), but the effect of cyproheptadine was relatively weak compared with chlorpheniramine. Moreover, BW A868C and ramatroban also showed the inhibition of nasal symptoms induced by the simultaneous administration of histamine and prostaglandin D(2) in a dose-dependent manner. BW A868C was more potent in inhibiting the nasal symptoms than ramatroban. These results clearly indicate that prostaglandin D(2) showed a synergistic effect on sneezing and nasal rubbing induced by histamine in rats, and its effect occurred through both prostaglandin D(2) and CRTH2 (chemoattractant receptor-homologous molecule expressed on TH2 cells) receptors. PMID- 17112501 TI - Ameliorative effect of pyrrolidinedithiocarbamate on acetic acid-induced colitis in rats. AB - Ulcerative colitis is a chronically recurrent inflammatory bowel disease of unknown origin. The present study examined the effect of NF-kappaB inhibitor and antioxidant, pyrrolidinedithiocarbamate (PDTC) on experimental ulcerative colitis in rats. Animals were randomly divided into 4 groups, each consisting of 6 animals; normal control group, acetic acid group, PDTC-treated group and sulfasalazine-treated group as a positive control group. Induction of colitis by intracolonic administration of 3% acetic acid produced severe macroscopic inflammation in the colon 24 h after acetic acid administration as assessed by the colonic damage score. Microscopically, colonic tissues showed ulceration, oedema and inflammatory cells infiltration. Biochemical studies revealed increased serum levels of lactate dehydrogenase (LDH), and nitrite/nitrate and colonic concentrations of tumor necrosis factor-alpha (TNF-alpha) and the neutrophil infiltration index, myeloperoxidase (MPO). Oxidative stress was indicated by elevated lipid peroxides formation and depleted reduced glutathione concentrations (GSH) in colonic tissues. Immunohistochemical studies of colonic sections revealed upregulation of inducible nitric oxide synthase (iNOS). Pretreatment with PDTC at a dose of (200 mg/kg/day, i.p.), three days before induction of colitis decreased serum LDH, nitrite/nitrate and TNF-alpha levels, colonic concentrations of MPO and lipid peroxides while increased colonic GSH concentration. Moreover, PDTC pretreatment attenuated colonic iNOS expression. Finally, histopathological changes were nearly restored by PDTC pretreatment. The findings of the present study provide evidence that PDTC may be beneficial in patients with inflammatory bowel disease. PMID- 17112502 TI - K201, a multi-channel blocker, inhibits clofilium-induced torsades de pointes and attenuates an increase in repolarization. AB - K201 (JTV519) is a 1,4-benzothiazepine derivative that exhibits a strong cardioprotective action and acts as a multiple-channel blocker, including as a K+ channel blocker. An experimental model of prolongation of the QT interval and torsades de pointes can be induced in rabbits by treatment with clofilium in the presence of the alpha1-adrenoreceptor agonist methoxamine. In this study we examined the effects of K201 with and without methoxamine on the QT and QTc intervals, and determined whether K201 inhibits clofilium-induced torsades de pointes in the presence of methoxamine (15 microg/kg/min) in rabbits (n=74). Administration of K201 (0, 40, 100, 200 and 400 microg/kg/min) with and without methoxamine prolonged the QT interval in a dose-dependent manner, and torsades de pointes did not occur in any animals. However, clofilium (50 microg/kg/min) with methoxamine induced torsades de pointes in all animals (6/6). Torsades de pointes occurred at rates of 100%, 67%, 40% and 0% at K201 concentrations of 0, 50, 200 and 400 microg/kg/min, respectively, in the clofilium-infused torsades de pointes model. Therefore, 400 microg/kg/min of K201 completely inhibited clofilium induced torsades de pointes and attenuated the increase of repolarization caused by clofilium; the inhibitory effects of K201 may be related to its pharmacological properties as an alpha1-adrenoceptor blocker. Overall, our results show that K201 causes prolongation of the QT and QTc intervals, but does not induce torsades de pointes, with and without alpha1-adrenoceptor stimulation. Furthermore, K201 inhibits clofilium-induced torsades de pointes, despite QT prolongation, suggesting that QT prolongation alone is not a proarrhythmic signal. PMID- 17112503 TI - Expression of brain derived neurotrophic factor, activity-regulated cytoskeleton protein mRNA, and enhancement of adult hippocampal neurogenesis in rats after sub chronic and chronic treatment with the triple monoamine re-uptake inhibitor tesofensine. AB - The changes of gene expression resulting from long-term exposure to monoamine antidepressant drugs in experimental animals are key to understanding the mechanisms of action of this class of drugs in man. Many of these genes and their products are either relevant biomarkers or directly involved in structural changes that are perhaps necessary for the antidepressant effect. Tesofensine is a novel triple monoamine reuptake inhibitor that acts to increase noradrenaline, serotonin, and dopamine neurotransmission. This study was undertaken to examine the effect of sub-chronic (5 days) and chronic (14 days) administration of Tesofensine on the expression of brain derived neurotrophic factor (BDNF) and activity-regulated cytoskeleton protein (Arc) in the rat hippocampus. Furthermore, hippocampi from the same animals were used to investigate the effect on cell proliferation by means of Ki-67- and NeuroD-immunoreactivity. We find that chronic, but not sub-chronic treatment with Tesofensine increases BDNF mRNA in the CA3 region of the hippocampus (35%), and Arc mRNA in the CA1 of the hippocampus (65%). Furthermore, the number of Ki-67- and neuroD-positive cells increased after chronic, but not sub-chronic treatment. This study shows that Tesofensine enhances hippocampal gene expression and new cell formation indicative for an antidepressant potential of this novel drug substance. PMID- 17112504 TI - Paradoxical hyperalgesia induced by mu-opioid receptor agonist endomorphin-2, but not endomorphin-1, microinjected into the centromedial amygdala of the rat. AB - The effects of endomorphin-2 or endomorphin-1 microinjected into the centromedial amygdala on the thermally-induced tail-flick response were studied in male CD rats. Microinjection of endomorphin-2 (8.7-35.0 nmol) given into the centromedial amygdala time- and dose-dependently decreased the tail-flick latencies. On the other hand, endomorphin-1 (8-32.6 nmol) given into the same site did not cause any change of the tail-flick latency. However, endomorphin-1 (32.6 nmol) or endomorphin-2 (35.0 nmol) given into the basolateral site of amygdala did not affect the tail-flick latency. Pretreatment with the antiserum against dynorphin A(1-17) (200 microg) significantly reversed the decrease of the tail-flick latency induced by endomorphin-2. The decrease of the tail-flick latency induced by endomorphin-2 was also blocked by the endomorphin-2 selective micro-opioid receptor antagonist 3-methoxynaltrexone (6.4 pmol) and by the N-methyl-D aspartate (NMDA) receptor antagonist MK-801 (30 nmol), but not by the kappa opioid receptor antagonist nor-binaltorphimine (6.6 nmol). It is concluded that endomorphin-2, but not endomorphin-1, given into the centromedial amygdala stimulates a 3-methoxynaltrexone-sensitive mu-opioid receptor subtype to induce the release of dynorphin A(1-17), which then acts on the NMDA receptor, but not kappa-opioid receptor for producing hyperalgesia. This conclusion is further supported by the additional findings that dynorphin A(1-17) (2.3 nmol) given into the centromedial amygdala also caused the decrease of the tail-flick latency, which was similarly blocked by the NMDA receptor antagonist MK-801 (30 nmol), but not kappa-opioid receptor antagonist nor-binaltorphimine (6.6 nmol). PMID- 17112505 TI - Intrathecally administered COX-2 but not COX-1 or COX-3 inhibitors attenuate streptozotocin-induced mechanical hyperalgesia in rats. AB - Members of the cyclooxygenase (COX) family are known to catalyze the rate limiting steps of prostaglandins synthesis and reported to be involved in neuropathic pain. Diabetic neuropathy is a type of neuropathic pain, though it is not clear if COX is relevant to the condition. Recently, spinal COX-2 protein was found to be increasing in streptozotocin-induced rats as compared to the constitutive expression. We attempted to determine which cyclooxygenase isoforms are involved in streptozotocin-induced mechanical hyperalgesia, which was induced by a single intraperitoneal injection of 75 mg/kg of streptozotocin. Intrathecal administrations of the COX-2 inhibitors SC-58125 (7-100 microg) and NS-398 (7-60 microg), as well as a high dose (100 microg) of the COX-1 inhibitor SC-560 attenuated hyperalgesia, whereas intrathecal administrations of a low dose (10 microg) of SC-560 and the COX-3 inhibitor acetaminophen (1-7 mg) did not. Further, intrathecal administration of SC-58125 (100 microg) did not produce an analgesic effect in normal rats. These results indicate that intrathecal administration of COX-2 inhibitors has an anti-hyperalgesic effect on streptozotocin-induced mechanical hyperalgesia and we concluded that spinal COX-2 is pivotal in streptozotocin-induced hyperalgesia. PMID- 17112506 TI - Simple-to-use, reference criteria for revealing drug-induced QT interval prolongation in conscious dogs. AB - Electrocardiogram (ECG) QT interval prolongation produced by drugs in certain animal models is currently believed to be predictive of cardiac proarrhythmic effects in humans. For this reason, nonclinical assessment of the effects of novel drugs on cardiac repolarization is a regulatory prerequisite for progressing such agents to clinical evaluation. The present investigation was carried out to develop reliable, simple-to-use reference criteria for identifying individual animals as responders to drugs that prolong the QT interval. ECG were recorded for 30 s at 0 (8 am), 2, 4, 6 and 24 h in 6 trained, conscious, beagle dogs during 5 control experimental sessions. QT intervals were measured and corrected for heart rate by applying the Van de Water algorithm (QTc). The maximal (QTc(max)) and minimal (QTc(min)) values of QTc observed in each of the five control recording sessions were noted. Two reference (R) criteria were used to designate an individual animal as a responder to drug treatment: 1) QTc(maxR) which was obtained by adding 10 ms to the largest value of QTc(max) observed during the five control recording sessions and 2) (QTc(max)-QTc(min))(maxR) which was obtained by increasing by 50% the largest of the (QTc(max)-QTc(min)) values [(QTc(max)-QTc(min))(max)] observed in the 5 control recording sessions. The sensitivity and reliability of these criteria were tested by determining QTc intervals before and 2, 4, 6 and 24 h after placebo or quinidine (200, 400 and 800 mg p.o. per animal). The reference values of QTc(maxR) and (QTc(max) QTc(min))(maxR) for the various dogs ranged from 246 to 270 ms and from 15 to 19.5 ms, respectively. The number of dogs responding to treatment (T: quinidine at 200, 400 and 800 mg, p.o. per animal) with a QTc(maxT) and/or a (QTc(max) QTc(min))(maxT) equal to or greater than the respective reference values was, respectively, 1/6, 3/6 and 5/6 dogs. Additionally, the number of responders correlated well with the concentration of free quinidine in the plasma. In conclusion, this investigation succeeded in establishing reliable, reference criteria for individual dogs despite the intrinsic daily variation of QTc interval. The application of these criteria allowed identifying individual animals responding to quinidine with delayed cardiac repolarization. PMID- 17112507 TI - 12(S)-HPETE induces itch-associated scratchings in mice. AB - The itch-associated responses evoked by intradermal injection of 12(S)-HPETE and leukotriene B4 were compared in ICR-mice. 12(S)-HPETE and leukotriene B4 (0.01 0.2 nmol/site) induced scratching of the injected site, respectively; the dose responses were a peak at 0.05 nmol/site (12(S)-HPETE) or 0.03 nmol/site (leukotriene B4). The scratching response by 12(S)-HPETE (0.05 nmol/site) started within 1 min, peaked in the first 10 min period, had almost subsided by 25 min whereas the effect of leukotriene B4 peaked in the second 10 min. The effect of leukotriene B4 is slightly stronger than that of 12(S)-HPETE in 40 min of count. The scratching induced by 12(S)-HPETE was inhibited by capsaicin, naltrexon, and LY255283. These results suggest the possibility that 12-lipoxygenase product can be added to a new member of an endogenous itch mediator in the skin. PMID- 17112508 TI - Sex differences and role of gonadal hormones on glutamate level in the nucleus accumbens in morphine tolerant rats: a microdialysis study. AB - Sex differences are observed in the development of tolerance to the antinociceptive effect of opioid drugs such as morphine, but the precise underlying mechanism remains unclear. There are evidences about the interaction between gonadal hormones and neuromodulatory systems including opioidergic and glutamatergic systems. We examined the sex differences and the role of gonadal hormones on the glutamate level in the nucleus accumbens in morphine tolerant rats using in vivo microdialysis. A microdialysis probe was implanted into the left nucleus accumbens core of rats and CSF (cerebrospinal fluid) dialysates were collected. The concentration of glutamate was measured by high-performance liquid chromatography with a fluorescence detector. The results showed that after chronic morphine administration, tolerance to antinociceptive effects of morphine was significantly greater in male rats (P<0.001). Sex differences in tolerance to morphine disappeared with gonadectomy of animals. There was also a significant sex difference in the glutamate level in the nucleus accumbens of morphine tolerant rats (P<0.001), ovariectomy of female rats decreased the glutamate level significantly (P<0.001), while gonadectomy did not change the glutamate level in males significantly. In conclusion, these experiments demonstrate that the excitatory amino acid release in the nucleus accumbens may be modulated by an estrogen-sensitive mechanism and play a role in the morphine analgesia and tolerance. PMID- 17112509 TI - Serotonin in microdialysate from the mediobasal hypothalamus increases after progesterone administration to estrogen primed macaques. AB - Estrogen and progesterone act on gene and protein expression in serotonin neurons in a manner that suggests serotonin neurotransmission should increase. However, measurement of extracellular serotonin in macaques was lacking. Elevated prolactin secretion can be an indicator of increased serotonergic function and prolactin is increased by combined estrogen and progesterone treatment. We examined extracellular serotonin by microdialysis in a well-characterized macaque model of steroid-induced prolactin secretion. Monkeys were fitted with 2 guide tubes directed to the arcuate nucleus of the hypothalamus. Samples (75 microl/15 minute interval) were obtained via a tether-swivel device through sample lines into an adjoining room. Serotonin was measured with a modified commercial enzyme linked immunoassay (ELISA) kit. Fenfluramine infused through the probe (300 microM for 2 h; n=2 trials) or administered intravenously (2.5 mg/kg; n=2 trials) caused a marked increase in extracellular serotonin and verified the efficacy of the procedure. Three monkeys were maintained with an estrogen implant for 2 weeks. Each monkey was injected with 20 mg of progesterone s.c. in oil at 1500 h; microdialysis was initiated the next morning and samples were obtained for 24 h. There was a significant increase in serotonin between 40 and 43 h after the progesterone injection (P<0.001, ANOVA). Serotonin averaged 59+/-1 pg/sample from 18-30 h post-progesterone injection, and averaged 76+/-2 pg/sample from 30-48 h post-progesterone injection (P<0.0001; t-test). Since the increase in serotonin is delayed by approximately 40 h after progesterone-injection, we speculate that the action of progesterone may involve either nuclear progestin receptors or membrane progestin receptors. PMID- 17112510 TI - Consequences of Shb and c-Abl interactions for cell death in response to various stress stimuli. AB - The adaptor protein Shb has previously been shown to regulate apoptosis in response to cytokines and inhibitors of angiogenesis although the mechanisms governing these effects have remained obscure. We currently demonstrate interactions between Shb and c-Abl and that Shb regulates c-Abl kinase activity. The data suggest that c-Abl binds to tyrosine phosphorylated Shb via a concerted effort involving both the c-Abl SH3 and SH2 domains. The biological significance of the Shb/c-Abl interaction was presently tested in overexpression experiments and was found to promote hydrogen peroxide-induced cell death. We also show by Shb knockdown experiments that Shb regulates c-Abl activity and modulates cell death in response to the genotoxic agent cisplatin and the endoplasmic reticulum stress-inducer tunicamycin. The findings are in agreement with the notion of Shb playing a pivotal role in modulating c-Abl pro-apoptotic signaling in response to various stress stimuli. PMID- 17112511 TI - Keratins modulate hepatic cell adhesion, size and G1/S transition. AB - Keratins (Ks) are the intermediate filament (IF) proteins of epithelial cells. Hepatocyte IFs are made solely of keratins 8 and 18 (K8/K18), the hallmark of all simple epithelia. While K8/K18 are essential for maintaining structural integrity, there is accumulating evidence indicating that they also exert non mechanical functions. We have reported recently that K8/K18-free hepatocytes from K8-null mice are more sensitive to Fas-mediated apoptosis, in line with an increased Fas density at the cell surface and an altered c-Flip regulation of the anti-apoptotic ERK1/2 signaling pathway. In the present study, we show that K8 null hepatocytes attach more rapidly but spread more slowly on a fibronectin substratum and undergo a more efficient G1/S transition than wild-type hepatocytes. Moreover, plectin, an IF associated protein, receptor for activated C kinase 1 (RACK1), a plectin partner, and vinculin, a key component of focal adhesions, distribute differently in spreading K8-null hepatocytes. Cell seeding leads to no differential activation of ERK1/2 in WT versus K8-null hepatocytes, whereas a stronger Akt activation is detected in K8-null hepatocytes. Insulin stimulation also leads to a differential Akt activation, implying altered Akt signaling capacity as a result of the K8/K18 loss. In addition, a delayed autophosphorylation of FAK, a target for integrin beta1 signaling, was obtained in seeding K8-null hepatocytes. These alterations in cell cycle-related events in hepatocytes in primary culture are also found in a K8-knockdown H4-II-E-C3 rat hepatoma cell line. Besides, K8/K18-free cells are smaller and exhibit a reduced rate of protein synthesis. In addition, a distinctive cyclin interplay is observed in these K8/K18-free hepatic cells, namely a more efficient cyclin A dependent G1/S phase transition. Furthermore, K8 re-expression in these cells, following transfer of a human K8 cDNA, restores proper cell size, spreading and growth. Together, these results suggest new interrelated signaling roles of K8/18 with plectin/RACK1 in the modulation of cell attachment/spreading, size/protein synthesis and G1/S transition. PMID- 17112512 TI - Auditory hair cell explant co-cultures promote the differentiation of stem cells into bipolar neurons. AB - Auditory neurons, the target neurons of the cochlear implant, degenerate following a sensorineural hearing loss. The goal of this research is to direct the differentiation of embryonic stem cells (SCs) into bipolar auditory neurons that can be used to replace degenerating neurons in the deafened mammalian cochlea. Successful replacement of auditory neurons is likely to result in improved clinical outcomes for cochlear implant recipients. We examined two post natal auditory co-culture models with and without neurotrophic support, for their potential to direct the differentiation of mouse embryonic SCs into characteristic, bipolar, auditory neurons. The differentiation of SCs into neuron like cells was facilitated by co-culture with auditory neurons or hair cell explants, isolated from post-natal day five rats. The most successful combination was the co-culture of hair cell explants with whole embryoid bodies, which resulted in significantly greater numbers of neurofilament-positive, neuron-like cells. While further characterization of these differentiated cells will be essential before transplantation studies commence, these data illustrate the effectiveness of post-natal hair cell explant co-culture, at providing valuable molecular cues for directed differentiation of SCs towards an auditory neuron lineage. PMID- 17112513 TI - Hypothermia in acute stroke--slow versus fast rewarming an experimental study in rats. AB - The rewarming phase after therapeutic hypothermia in cerebral ischemia appears crucial as rapid rewarming may lead to rebound phenomena and enhance deleterious ischemic effects. We hypothesized that slow and controlled rewarming after moderate hypothermia is superior to fast rewarming in rats subjected to 90 min temporary middle cerebral artery occlusion (tMCAO). Two experiments were designed: (i) 34 rats were randomly assigned to either normothermic treatment, to hypothermia (33 degrees C) with rapid rewarming within 20 min, or to hypothermia with slow rewarming within 2 h after 4 h of hypothermia starting 2 h after tMCAO. Infarct size, neuroscore, myeloperoxidase and aquaporin 4 (AQP4) positive cells were assessed on day 5 after tMCAO. (ii) In 15 rats, striatal cerebral microdialysis was performed from 1.5 h before until 8 h after tMCAO. Total infarct volume was largest in the normothermic group (89.9+/-16.8 mm(3)) followed by the fast rewarming group (69.2+/-12.6 mm(3)), and a significantly smaller infarct volume in the slow rewarming group (41.1+/-6.6 mm(3), p<0.05). Neurological functions improved in both hypothermia groups at day 5 after tMCAO (Neuroscore median 2.5 in normothermia vs. 1.5 in both hypothermia groups) though without any difference between slowly and fast rewarmed animals. Periinfarct expression of AQP4 was less prominent in slowly rewarmed animals as was the count of MPO-positive cells in subcortical regions. Glutamate release was significantly higher at 4 distinct time points in the control group. Slow rewarming after a period of hypothermia is superior to fast rewarming. It may blunt deleterious rebound effects such as overexpression of AQP4, sustain anti-inflammatory mechanisms and thereby preserve the neuroprotection delivered by hypothermia. PMID- 17112514 TI - Actions of neuropoietic cytokines and cyclic AMP in regenerative conditioning of rat primary sensory neurons. AB - A conditioning lesion to peripheral axons of primary sensory neurons accelerates regeneration of their central axons in vivo or neurite outgrowth if the neurons are grown in vitro. Previous evidence has implicated neuropoietic cytokines and also cyclic AMP in regenerative conditioning. In experiments reported here, delivery through a lentivirus vector of ciliary neurotrophic factor to the appropriate dorsal root ganglion in rats was sufficient to mimic the conditioning effect of peripheral nerve injury on the regeneration of dorsal spinal nerve root axons. Regeneration in this experimental preparation was also stimulated by intraganglionic injection of dibutyryl cyclic AMP but the effects of ciliary neurotrophic factor and dibutyryl cyclic AMP were not additive. Dibutyryl cyclic AMP injection into the dorsal root ganglion induced mRNAs for two other neuropoietic cytokines, interleukin-6 and leukemia inhibitory factor and increased the accumulation of phosphorylated STAT3 in neuronal nuclei. The in vitro conditioning action of dibutyryl cyclic AMP was partially blocked by a pharmacological inhibitor of Janus kinase 2, a neuropoietic cytokine signaling molecule. We suggest that the beneficial actions of increased cyclic AMP activity on axonal regeneration of primary sensory neurons are mediated, at least in part, through the induction of neuropoietic cytokine synthesis within the dorsal root ganglion. PMID- 17112515 TI - In vivo visualization of focal demyelination in peripheral nerves by gadofluorine M-enhanced magnetic resonance imaging. AB - Magnetic resonance imaging (MRI) allows assessment of axonal nerve lesions, but detection of focal demyelination is still difficult. We have recently shown that the novel micellar magnetic resonance (MR) contrast agent gadofluorine M (Gf) accumulates in nerve fibers undergoing Wallerian degeneration. In the present study, we report on the in vivo visualization of focal demyelination induced by lysolecithin. Upon appropriate intraneural injection, lysolecithin focally dissolves myelin sheaths with sparing of axons. Conventional unenhanced and gadolinium-DTPA enhanced T1-w MRI did not show signal alterations or contrast enhancement. In contrast, application of Gf led to bright contrast enhancement on T1-w images at the site of focal demyelination, but spared distal nerve segments not affected by demyelination. Gf enhancement persisted until remyelination had occurred. Our study shows that areas of focal nerve demyelination can be detected in vivo by Gf-enhanced MRI. This finding opens up a broad spectrum of applications in experimental neurology, and, depending on further clinical development of Gf, may aid in the diagnostic work up of patients with patchy, multifocal demyelinative disorders in the future. PMID- 17112516 TI - Aging of the rat mesostriatal system: differences between the nigrostriatal and the mesolimbic compartments. AB - The impairment of the mesostriatal dopaminergic system has been considered responsible for motor and affective disturbances associated with aging and a risk factor for Parkinson's disease. However, the basic mechanisms underlying this phenomenon are still unknown. Here we used biochemical, molecular and morphological techniques directed at detecting flaws in the dopamine synthesis route and signs of dopaminergic degeneration in the rat mesostriatal system during normal aging. We found two different age-related processes. One is characterized by a dopa decarboxylase decrease, and involves both the nigrostriatal and mesolimbic compartments, and is responsible for a moderate dopamine loss in the dorsal striatum, where other parameters of dopamine synthesis are not affected. The other is characterized by axonal degeneration with aggregation of phosphorylated forms of tyrosine hydroxylase (TH) and amyloid precursor protein in degenerate terminals, and alpha-synuclein in their original somata. This process is restricted to mesolimbic regions and is responsible for the decline of TH activity and l-dopa levels and the greater decrease in dopamine levels in this compartment. These findings suggest that both the nigrostriatal and the mesolimbic systems are vulnerable to aging, but in contrast to what occurs in Parkinson's disease, the mesolimbic system is more vulnerable to aging than the nigrostriatal one. PMID- 17112517 TI - Amyloid-beta peptide affects viability but not differentiation of embryonic and adult rat hippocampal progenitor cells. AB - The neurological deficits that are characteristic of Alzheimer's Disease (AD) are ultimately a result of neuronal loss in distinct anatomical regions of the brain. This neuronal loss is thought to be due, in large part to the presence of the neurotoxic beta-amyloid (Abeta) deposits, that are characteristic of the AD brain. Transplantation therapy, in which neural stem cells (NSCs) or neural progenitor cells (NPCs) are introduced into damaged regions of the brain and induced to differentiate into replacement neurons, has been proposed as a possible therapeutic approach to treat AD. However, in the AD brain Abeta plaques, which remain in the area of neuronal degeneration, may affect the viability or differentiation potential of transplanted NSCs. Currently there is contradictory evidence concerning the effect of Abeta on NSCs. To further investigate the effect of Abeta on NSCs, we compared the mitochondrial function, proliferation and cellular differentiation of two populations of hippocampal NSCs (embryonic and adult derived) after Abeta exposure. Our results highlight the heterogeneity between different populations of NSCs even when derived from the same brain region. Our data also demonstrate that while mitochondrial function of NSCs is affected by Abeta, their proliferation and differentiation are not significantly influenced. Considered with previous studies, our results suggest that while NSCs do respond to the presence of Abeta, proliferation and differentiation of certain populations are not affected. Further study of the differences between susceptible vs. resistant populations of NSCs may provide crucial clues for the development of effective therapies to combat AD. PMID- 17112518 TI - Cortical reorganization in NT3-treated experimental spinal cord injury: Functional magnetic resonance imaging. AB - Functional magnetic resonance imaging (fMRI) studies were performed for visualizing ongoing brain plasticity in Neurotrophin-3 (NT3)-treated experimental spinal cord injury (SCI). In response to the electrical stimulation of the forepaw, the NT3-treated animals showed extensive activation of brain structures that included contralateral cortex, thalamus, caudate putamen, hippocampus, and periaqueductal gray. Quantitative analysis of the fMRI data indicated significant changes both in the volume and center of activations in NT3-treated animals relative to saline-treated controls. A strong activation in both ipsi- and contralateral periaqueductal gray and thalamus was observed in NT3-treated animals. These studies indicate ongoing brain reorganization in the SCI animals. The fMRI results also suggest that NT3 may influence nociceptive pathways. PMID- 17112519 TI - Long-term feeding a high-fat diet causes histological and parasitological effects on murine schistosomiasis mansoni outcome. AB - This study investigated whether long-term feeding a high-fat diet (HFC) has an effect on schistosomiasis mansoni outcome compared to standard chow diet (SC). Swiss Webster female mice (3 wk old) fed each diet over 5 months, and then were infected with 50 Schistosoma mansoni cercariae. Their nutritional status was assessed by monitoring growth rates twice a week and measuring serum levels of lipoproteins. Mice were euthanised 63 days after infection. Parasitological and liver histological analyses were performed. The levels of TC, HDL-C and LDL-C, fecal and tissue schistosome eggs were statistically different (p<0.05) between groups. Livers from HFC mice showed exudative, exudative/exudative-productive, exudative-productive and productive granulomas, some degree of hepatic steatosis and focal necrosis. Mice fed normal-chow did not present productive granulomas and hepatic steatosis. The morphometric evaluation of hepatic granulomas did not reach statistical significance (p>0.05) between diets assayed. The high-fat diet for long-term produces effects on schistosomiasis mansoni outcome. PMID- 17112520 TI - Aluminum inhibits proteolytic degradation of amyloid beta peptide by cathepsin D: a potential link between aluminum accumulation and neuritic plaque deposition. AB - Neuritic plaques are the key pathological feature of Alzheimer's disease, and amyloid beta (Abeta) peptides are major component of these plaques. In this study, we demonstrated the influence of aluminum (Al) on the Abeta peptide degradation by cathepsin D. Al did not directly affect the cathepsin D activity using small synthetic substrate. However, when Abeta peptides were used as substrate, the apparent inhibitory effect of Al on cathepsin D activity was observed. This inhibitory effect disappeared by treatment of desferrioxamine. These results indicate that Al has the potential to interact and disrupt Abeta peptide catabolism via the inhibition of proteolytic degradation. PMID- 17112521 TI - Gain- and loss-of-function mutations in Zat10 enhance the tolerance of plants to abiotic stress. AB - C(2)H(2)-zinc finger proteins that contain the EAR repressor domain are thought to play a key role in modulating the defense response of plants to abiotic stress. Constitutive expression of the C(2)H(2)-EAR zinc finger protein Zat10 in Arabidopsis was found to elevate the expression of reactive oxygen-defense transcripts and to enhance the tolerance of plants to salinity, heat and osmotic stress. Surprisingly, knockout and RNAi mutants of Zat10 were also more tolerant to osmotic and salinity stress. Our results suggest that Zat10 plays a key role as both a positive and a negative regulator of plant defenses. PMID- 17112522 TI - Characterization of red cell membrane proteins as a function of red cell density: annexin VII in different forms of hereditary spherocytosis. AB - Fresh human blood samples were collected from healthy controls and splenectomized and unsplenectomized patients with hereditary spherocytosis due to band 3 or ankyrin and spectrin deficiency. The erythrocytes were separated into age-related fractions using self-forming Percoll density gradients. Membrane proteins were analysed by 2D electrophoresis and identified by mass spectrometry. Annexin VII was present in reticulocytes but was then lost as the cells matured. A different pattern was found in band 3-deficient samples: annexin VII was in fact present in both mature and immature red cell membranes. Cytoskeletal anomalies may then influence the turn-over of annexin VII during erythrocyte maturation. PMID- 17112523 TI - Correlates of depression in type 2 diabetic elderly patients: a correlational study. AB - OBJECTIVES: This aim of this study was to investigate depression and related factors in elderly patients (65 years) with type 2 diabetics. METHODS: The study utilized a descriptive-correlational design. Convenience sampling was applied to enroll 156 subjects, aged >or= 65 years from diabetic outpatient clinics at three hospitals in Kaohsiung. This study applied the Personal Resource Questionnaire 2000 (PRQ 2000), Taiwan Geriatric Depression Scale (TGDS), and a form for demographic data. RESULTS: (1) The mean index score for depression level was 26, indicating that study subjects had a low level of depression. The social support index score was 76. (2) Significant differences in TGDS scores existed for diabetes duration, diabetes complications, and treatment type. (3) Social support and regular exercise were significantly and negatively correlated with depression. (4) Social support and diabetes complications were significant correlates of depression and accounted for 25.9% of variance in depression. CONCLUSION: Analytical results may assist nurses in understanding depression and related factors for diabetic patients aged >or= 65. Therefore, this study could form a basis for caring older people with diabetes, and provide a reference for further research. PMID- 17112524 TI - Differential expression of Paragonimus westermani eggshell proteins during the developmental stages. AB - Eggs of trematode parasites are comprised of numerous vitelline cells and one fertilized ovum, and are encapsulated within a protein shell provided by the vitellocytes. In this study, we isolated two full-length cDNA clones that showed substantial levels of sequence identity with trematode-specific eggshell precursor proteins from the human lung fluke, Paragonimus westermani. These cDNAs, designated Pw-Vit20 (868-bp-long) and Pw-Vit36 (883-bp-long), shared a 76% identity with one another at the nucleotide level, and each encoded a 261-amino acid (aa) polypeptide. The deduced aa sequences contained a N-terminal hydrophobic segment, as well as a sequence motif of Gly-Gly-Gly-Tyr-Asp-Asn/Thr Tyr-Gly-Lys/Gln, which is highly homologous with the eggshell proteins of Fasciola hepatica. With the high frequencies of tyrosine, glycine and lysine, the positions occupied by tyrosine, which has been proved to be converted into dihydroxyphenylalanine, were well preserved. Pw-Vit20 and Pw-Vit36 were found to be monoexonic genes with variably diverged variants scattered into multiple genomic loci. Their protein products were localized in the vitelline follicles and eggshells. Expression of Pw-Vit20 was restricted to the egg and adult stages, thus suggesting a critical involvement of Pw-Vit20 in the parasite's fecundity activity. Conversely, Pw-Vit36 was constitutively expressed in the metacercariae and juvenile stages in the vitelline follicles and ducts, which suggested that the prepositioning of stem or primordial vitelline cells within the juveniles prior to sexual maturation. Pw-Vit36 might acquire a unique or additional function relevant to the maturation and/or development of the vitelline cells/follicles during the evolutionary period of P. westermani. Differential biological implications of multiple eggshell precursor proteins may provide insight into the molecular mechanism of eggshell formation and the developmental process of the vitelline follicles in the parasitic trematode. PMID- 17112525 TI - Differential gene expression in hypobiosis-induced and non-induced third-stage larvae of the bovine lungworm Dictyocaulus viviparus. AB - Hypobiosis is of particular importance in overwintering of the bovine lungworm Dictyocaulus viviparus. However, in parasitic nematodes there is no information available on the genetic mechanisms of hypobiosis. Suppression subtractive hybridisation was performed to identify upregulated transcripts of hypobiosis induced and non-induced third-stage D. viviparus larvae, respectively. Subtracted libraries containing 105 clones of the hypobiosis-induced and 104 clones of the non-induced larvae were generated. By differential screening and Southern dot blot, 26 clones of the hypobiosis-induced and 22 clones of the non-induced larvae were confirmed to be differentially expressed. Sequencing of rapid amplification of cDNA ends (RACE) and spliced-leader-1 PCR products was performed to further characterise selection of the differentially regulated gene transcripts. The genes encoding an N-methyltransferase and a superoxide dismutase were upregulated in the hypobiosis-induced and non-induced larvae, respectively. The expression patterns of these genes were validated by quantitative real-time PCR. This revealed differential gene expression, particularly for the N-methyltransferase. PMID- 17112526 TI - Alternative invasion pathways for Plasmodium berghei sporozoites. AB - Invasion of hepatocytes by Plasmodium sporozoites is a prerequisite for establishment of a natural malaria infection. The molecular mechanisms underlying sporozoite invasion are largely unknown. We have previously reported that infection by Plasmodium falciparum and Plasmodium yoelii sporozoites depends on CD81 and cholesterol-dependent tetraspanin-enriched microdomains (TEMs) on the hepatocyte surface. Here we have analyzed the role of CD81 and TEMs during infection by sporozoites from the rodent parasite Plasmodium berghei. We found that depending on the host cell type, P. berghei sporozoites can use several distinct pathways for invasion. Infection of human HepG2, HuH7 and HeLa cells by P. berghei does not depend on CD81 or host membrane cholesterol, whereas both CD81 and cholesterol are required for infection of mouse hepatoma Hepa1-6 cells. In primary mouse hepatocytes, both CD81-dependent and -independent mechanisms participate in P. berghei infection and the relative contribution of the different pathways varies, depending on mouse genetic background. The existence of distinct invasion pathways may explain why P. berghei sporozoites are capable of infecting a wide range of host cell types in vitro. It could also provide a means for human parasites to escape immune responses and face polymorphisms of host receptors. This may have implications for the development of an anti malarial vaccine targeting sporozoites. PMID- 17112527 TI - Enzymes of type II fatty acid synthesis and apicoplast differentiation and division in Eimeria tenella. AB - Apicomplexan parasites, Eimeria tenella, Plasmodium spp. and Toxoplasma gondii, possess a homologous plastid-like organelle termed the apicoplast, derived from the endosymbiotic enslavement of a photosynthetic alga. However, currently no eimerian nuclear encoded apicoplast targeted proteins have been identified, unlike in Plasmodium spp. and T. gondii. In this study, we demonstrate that nuclear encoded enoyl reductase of E. tenella (EtENR) has a predicted N-terminal bipartite transit sequence, typical of apicoplast-targeted proteins. Using a combination of immunocytochemistry and EM we demonstrate that this fatty acid biosynthesis protein is located in the apicoplast of E. tenella. Using the EtENR as a tool to mark apicoplast development during the Eimeria lifecycle, we demonstrate that nuclear and apicoplast division appear to be independent events, both organelles dividing prior to daughter cell formation, with each daughter cell possessing one to four apicoplasts. We believe this is the first report of multiple apicoplasts present in the infectious stage of an apicomplexan parasite. Furthermore, the microgametes lacked an identifiable apicoplast consistent with maternal inheritance via the macrogamete. It was found that the size of the organelle and the abundance of EtENR varied with developmental stage of the E. tenella lifecycle. The high levels of EtENR protein observed during asexual development and macrogametogony is potentially associated with the increased synthesis of fatty acids required for the rapid formation of numerous merozoites and for the extracellular development and survival of the oocyst. Taken together the data demonstrate that the E. tenella apicoplast participates in type II fatty acid biosynthesis with increased expression of ENR during parasite growth. Apicoplast division results in the simultaneous formation of multiple fragments. The division mechanism is unknown, but is independent of nuclear division and occurs prior to daughter formation. PMID- 17112528 TI - The differential associations between HDL, non-HDL and total cholesterols and atherosclerotic calcium deposits in multiple vascular beds. AB - BACKGROUND: The purpose of this study was to determine the magnitude of associations between four different cholesterol measures and systemic atherosclerotic calcification. METHODS: One thousand and seventy-eight consecutive patients were evaluated by electron beam computed tomography for the extent of calcified atherosclerosis in the carotids, coronaries, thoracic aorta, abdominal aorta, renals and iliacs, as well as the mitral and aortic annuli. HDL, non-HDL and total cholesterol were measured using the Cholestec LDX system. RESULTS: Overall, the prevalence of calcium in the carotids, coronaries, thoracic aorta, abdominal aorta, iliacs, renals, mitral annulus and aortic annulus was 31.6, 58.7, 40.5, 55.0, 16.8, 57.0, 8.3 and 23.6%, respectively. After multivariable adjustment, a 1-S.D. interval in the total to HDL cholesterol ratio in men was significantly associated with 36 and 29% higher odds for any calcium in the abdominal aorta and iliac arteries, respectively. In women, this same interval was significantly associated with odds ratios of 1.66 and 1.41 for the presence of any calcium in the thoracic and abdominal aorta, respectively. CONCLUSION: Measurement of HDL and non-HDL cholesterols, as well as calculation of the total cholesterol to HDL ratio, may provide a practical means for stratifying the risk for extra-coronary calcified atherosclerosis. PMID- 17112529 TI - Pravastatin improved glucose metabolism associated with increasing plasma adiponectin in patients with impaired glucose tolerance and coronary artery disease. AB - Reduced incidence of type-2 diabetes has been shown in patients treated with pravastatin. Adiponectin can exhibit beneficial effects on glucose metabolism. We investigated whether pravastatin could improve glucose tolerance associated with increasing adiponectin levels in patients with impaired glucose tolerance (IGT). This study consisted of 40 coronary artery disease (CAD) patients with IGT assessed by oral glucose tolerance test (OGTT). Patients were randomized to receive pravastatin (n=20) or no lipid-lowering medications (control group, n=20) for 6 months, after which OGTT was repeated and adiponectin levels were measured. Pravastatin treatment significantly decreased levels of total cholesterol (16%), low-density lipoprotein cholesterol (23%) and high-sensitivity C-reactive protein (37%) (p<0.01, respectively). At 2h in OGTT, pravastatin significantly improved hyperglycemia (-14%) and hyperinsulinemia (-23%). Pravastatin treatment significantly elevated plasma adiponectin levels (35%; p<0.001) but not in the control group. The glucose reduction at 2h post-OGTT was significantly associated with increased levels of adiponectin (r=-0.462; p=0.003). Pravastatin treatment is an independent predictor for improvement of post-loaded hyperglycemia (odds ratio; 5.7; 95% confidence interval 1.7-19.3; p=0.003) and achieved beneficial conversion from IGT to normal glucose tolerance (40%; p=0.03). Pravastatin exhibits beneficial effects on glucose metabolism especially in the postprandial state associated with increasing plasma adiponectin levels in CAD patients with IGT. PMID- 17112530 TI - Quantitative trait locus on Chromosome 19 for circulating levels of intercellular adhesion molecule-1 in Mexican Americans. AB - Circulating soluble intercellular adhesion molecule-1 (sICAM-1) is a biochemical marker of inflammation. We performed variance-components-based quantitative genetic analyses in SOLAR of sICAM-1 in 1170 individuals from Mexican American families in the San Antonio Family Heart Study. The trait is heritable (h(2)=0.50+/-0.06, P<10(-6)). Multipoint linkage analysis using a approximately 10-cM microsatellite map revealed a region on Chromosome 19p near marker D19S586 showing strong evidence of linkage for sICAM-1 (empirically adjusted univariate equivalent LOD=4.95), coincident with the structural gene ICAM1. This region has been identified previously as a QTL for inflammatory, autoimmune, and metabolic syndrome traits. There is significant evidence (P=0.0023) of locus heterogeneity for sICAM-1 in this sample: a subset of pedigrees contributes most of the linkage signal for sICAM-1 on Chromosome 19, suggesting a logical focus for future genetic dissection of the trait. PMID- 17112531 TI - Mechanics and deformation of the nucleus in micropipette aspiration experiment. AB - Robust biomechanical models are essential for the study of nuclear mechanics and deformation and can help shed light on the underlying mechanisms of stress transition in nuclear elements. Here, we develop a computational model for an isolated nucleus undergoing micropipette aspiration. Our model includes distinct components representing the nucleoplasm and nuclear envelope. The nuclear envelope itself comprises three layers: inner and outer nuclear membranes and one thicker layer representing the nuclear lamina. The nucleoplasm is modeled as a viscoelastic Maxwell material with a single time constant, while a modified Maxwell model, equivalent to a spring and a dashpot in series and both in parallel with a spring, is adopted for the inner and outer nuclear membranes. The nuclear envelope layer is taken as a linear elastic material. The proposed computational model, validated using experimental observations of Guilak et al. [2000. Viscoelastic properties of the cell nucleus. Biochemical and Biophysical Research Communications 269, 781-786] and Deguchi et al. [2005, Flow-induced hardening of endothelial nucleus as an intracellular stress-bearing organelle. Journal of Biomechanics 38, 1751-1759], is employed to study nuclear mechanics and deformation in micropipette aspiration and to shed light on the contribution of individual nuclear components on the response. The results indicate that the overall response of an isolated nucleus in micropipette aspiration is highly sensitive to the apparent stiffness of the nuclear lamina. This observation suggests that micropipette aspiration is an effective technique for examining the influence of various kinds of alteration in the nuclear lamina, such as mutations in the gene encoding lamin A, and also structural remodeling due to mechanical perturbation. PMID- 17112532 TI - Bone regeneration during distraction osteogenesis: mechano-regulation by shear strain and fluid velocity. AB - Corroboration of mechano-regulation algorithms is difficult, partly because repeatable experimental outcomes under a controlled mechanical environment are necessary, but rarely available. In distraction osteogenesis (DO), a controlled displacement is used to regenerate large volumes of new bone, with predictable and reproducible outcomes, allowing to computationally study the potential mechanisms that stimulate bone formation. We hypothesized that mechano-regulation by octahedral shear strain and fluid velocity can predict the spatial and temporal tissue distributions seen during experimental DO. Variations in predicted tissue distributions due to alterations in distraction rate and frequency could then also be studied. An in vivo ovine tibia experiment evaluating bone-segment transport (distraction, 1 mm/day) over an intramedullary nail was used for comparison. A 2D axisymmetric finite element model, with a geometry originating from the experimental data, was created and included into a previously developed model of tissue differentiation. Cells migrated and proliferated into the callus, differentiating into fibroblasts, chondrocytes or osteoblasts, dependent on the biophysical stimuli. Matrix production was modelled with an osmotic swelling model to allow tissues to grow at individual rates. The temporal and spatial tissue distributions predicted by the computational model agreed well with those seen experimentally. In addition, it was observed that decreased distraction rate (0.5 mm/d vs. 0.25 mm/d) increased the overall time needed for complete bone regeneration, whereas increased distraction frequency (0.5 mm/12 h vs. 0.25 mm/6 h) stimulated faster bone regeneration, as found in experimental findings by others. Thus, the algorithm regulated by octahedral shear strain and fluid velocity was able to predict the bone regeneration patterns dependent on distraction rate and frequency during DO. PMID- 17112533 TI - Identification, characterization and quantitation of pyrogenic polycylic aromatic hydrocarbons and other organic compounds in tire fire products. AB - On 15 August 2001, a tire fire took place at the Pneu Lavoie Facility in Gatineau, Quebec, in which 4000 to 6000 new and recycled tires were stored along with other potentially hazardous materials. Comprehensive gas chromatography-mass spectrometry (GC-MS) analyses were performed on the tire fire samples to facilitate detailed chemical composition characterization of toxic polycyclic aromatic hydrocarbons (PAHs) and other organic compounds in samples. It is found that significant amounts of PAHs, particularly the high-ring-number PAHs, were generated during the fire. In total, 165 PAH compounds including 13 isomers of molecular weight (MW) 302, 10 isomers of MW 278, 10 isomers of MW 276, 7 isomers of MW 252, 7 isomers of MW 228, and 8 isomers of MW 216 PAHs were positively identified in the tire fire wipe samples for the first time. Numerous S-, O-, and N-containing PAH compounds were also detected. The identification and characterization of the PAH isomers was mainly based on: (1) a positive match of mass spectral data of the PAH isomers with the NIST authentic mass spectra database; (2) a positive match of the GC retention indices (I) of PAHs with authentic standards and with those reported in the literature; (3) agreement of the PAH elution order with the NIST (US National Institute of Standards and Technology) Standard Reference Material 1597 for complex mixture of PAHs from coal tar; (4) a positive match of the distribution patterns of PAH isomers in the SIM mode between the tire fire samples and the NIST Standard Reference Materials and well-characterized reference oils. Quantitation of target PAHs was done on the GC-MS in the selected ion monitoring (SIM) mode using the internal standard method. The relative response factors (RRF) for target PAHs were obtained from analyses of authentic PAH standard compounds. Alkylated PAH homologues were quantitated using straight baseline integration of each level of alkylation. PMID- 17112534 TI - Enantiomeric separation of novel anticancer agent 5-hydroxy-3-(4-methoxyphenyl)-2 (3,4,5-trimethoxyphenyl)-cyclopent-2-en-1-one. AB - The enantiomers of 5-hydroxy-3-(4-methoxyphenyl)-2-(3,4,5-trimethoxyphenyl) cyclopent-2-en-1-one, a novel anticancer agent, were separated by derivatisation with caronaldehyde, separation of the resulting diastereoisomers of the corresponding esters by silica gel column chromatography and regeneration of alcohols (S)-5-hydroxy-3-(4-methoxyphenyl)-2-(3,4,5-trimethoxyphenyl)-cyclopent-2 en-1-one and (R)-5-hydroxy-3-(4-methoxyphenyl)-2-(3,4,5-trimethoxyphenyl) cyclopent-2-en-1-one under aqueous conditions. The absolute configuration of the enantiomers was determined by 1H NMR studies of the corresponding Mosher esters. Alternatively, the enantiomers were separated by preparative HPLC to collect the (S)- and (R)-5-hydroxy-3-(4-methoxyphenyl)-2-(3,4,5-trimethoxyphenyl)-cyclopent-2 en-1-ones with high purity which was comparable with that obtained by the chemical method. The details of these methods have been presented herein. PMID- 17112535 TI - Multi-phase equilibrium microemulsions and synthesis of hierarchically structured calcium carbonate through microemulsion-based routes. AB - Middle-phase microemulsions (MPMs) in two systems of a cationic surfactant, tetradecyltrimethylammonium bromide (TTABr)/n-butanol/iso-octane/Na2CO3 or CaCl2 and an anionic surfactant, sodium dodecyl sulfate (SDS)/n-butanol/iso octane/Na2CO3 or CaCl2, were used to synthesize nanostructured calcium carbonates. MPMs provide a simple and versatile reaction media, i.e., upper-phase W/O, BC, and O/W structured equilibrium microemulsions to be used for synthesizing hierarchically structured CaCO3 at the nanometer scale. On the basis of the investigations on the phase behavior of the MPMs, hierarchically structured calcium carbonates with dendrites, ellipsoids, square-schistose cubes, and spheres were synthesized through the MPM-based routes. PMID- 17112536 TI - Detection of anti-heat shock protein 90 beta (Hsp90beta) antibodies in cerebrospinal fluid. AB - Antibodies against heat shock protein 90 beta (Hsp90beta) recognize the antigen on the cell surface of the oligodendrocyte precursor cells and cause a decrease of oligodendrocyte population in cell cultures. These antibodies have been found in patients with multiple sclerosis (MS). This report describes an original and sensitive method to detect anti-Hsp90beta antibodies in cerebrospinal fluid (CSF) using a western blot procedure. We have developed the method for autoantibody detection using Hsp90beta from cell membrane fraction instead of commercial Hsp90beta as antigen. The presence of anti-Hsp90beta antibodies in CSF of MS patients may play a pathogenic role in MS, and a large-scale study is needed to establish a possible diagnostic value of these antibodies in MS patients. PMID- 17112537 TI - Prevalence of Thelohania solenopsae infected Solenopsis invicta newly mated queens within areas of differing social form distributions. AB - Newly mated queens (NMQs) originating from monogyne red imported fire ant (Solenopsis invicta) colonies and following a mating flight, initiate new colonies by sealing themselves in a nuptial chamber and relying solely on their own fat and crop reserves, as well as no longer needed wing muscles to rear their first workers (claustral colony foundation). This method of colony-founding is rarely successful for polygyne-derived NMQs, whose low weight critically limits the number of first workers they are able to produce. However, this observation may be confounded by the parasitic microsporidium, Thelohania solenopsae, thus far found to persist only in association with polygyne colonies. Infections of this microsporidium reduce the weight of female alates and may explain why polygyne NMQs are unlikely to successfully found colonies claustrally. NMQs collected following mating flights in Gainesville and Ocala, Florida were sorted by weight, checked for insemination and T. solenopsae infection. Insemination levels were greater than 90% for all weight classes at both collection sites and were not related to infection. Infection levels were lower in Gainesville than Ocala, averaging 1.67% and 14.14%, respectively. Polygyne-derived NMQs collected in Ocala, defined here as weighing 12mg (social form correctly assigned in 85% of samples examined by PCR), had the highest infection levels, 25.37% (17/67) in 2003 and 21.43% (6/28) in 2004. We conclude that infection by T. solenopsae cannot be completely responsible for the inability of polygyne NMQs to claustrally establish colonies. PMID- 17112538 TI - Evidence for cystic fibrosis transmembrane conductance regulator chloride current in swine ventricular myocytes. AB - The present study investigated whether cAMP-dependent cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel current (i.e., I(Cl.CFTR) or I(Cl.cAMP)) would be expressed in pig cardiac myocytes using whole cell patch technique and reverse transcription polymerase chain reaction (RT PCR). It was found that the beta-adrenoceptor agonist isoproterenol activated a time-independent current in myocytes from the ventricle, but not the atrium of pig heart. Histamine and forskolin (an adenylate cyclase activator) induced a similar current in pig ventricular cells. The current induced by isoproterenol was blocked by the PKA inhibitor H-7, reduced by the replacement of external Cl( ) ion, and inhibited by the application of 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB), but not 4'-diisothiocynatostilbene-2,2'-disulfonic acid (DIDS), typical of I(Cl.CFTR). I(Cl.CFTR) showed a small difference in regional myocytes across the left ventricular wall from epicardium to endocardium. Isoproterenol-induced current was 3.1+/-0.2 (n=33), 2.8+/-0.2 (n=25) and 2.3+/ 0.2 pA/pF (n=31) respectively in subepicardial, midmyocardial, and subendocardial myocytes (P<0.05, subepicardium vs. subendocardium). RT-PCR and Western blotting analysis revealed that significant differences in CFTR channel mRNA and protein levels were present in atrial and ventricular cells, but not in regional ventricular cells across the ventricular wall from subepicardium to subendocardium. These results indicate that the functional CFTR channel (i.e., I(Cl.CFTR)) is present in ventricular myocytes, but not in atrial cells of pig heart. PMID- 17112539 TI - Influence of the internal disulfide bridge on the folding pathway of the CL antibody domain. AB - Disulfide bridges are one of the most important factors stabilizing the native structure of a protein. Whereas the basis for their stabilizing effect is well understood, their role in a protein folding reaction still seems to require further attention. We used the constant domain of the antibody light chain (C(L)), a representative of the ubiquitous immunoglobulin (Ig)-superfamily, to delineate the kinetic role of its single buried disulfide bridge. Independent of its redox state, the monomeric C(L) domain adopts a typical Ig-fold under native conditions and does not retain significant structural elements when unfolded. Interestingly, its folding pathway is strongly influenced by the disulfide bridge. The more stable oxidized protein folds via a highly structured on-pathway intermediate, whereas the destabilized reduced protein populates a misfolded off pathway species on its way to the native state. In both cases, the formation of the intermediate species is shown to be independent of the isomerization state of the Tyr(141)-Pro(142) bond. Our results demonstrate that the internal disulfide bridge in an antibody domain restricts the folding pathway by bringing residues of the folding nucleus into proximity thus facilitating the way to the native state. PMID- 17112540 TI - Adolescent anorexia nervosa: cross-sectional and follow-up frontal gray matter disturbances detected with proton magnetic resonance spectroscopy. AB - There are very few magnetic resonance spectroscopy studies in anorexia nervosa and none of them with young adolescent patients. We studied 12 anorexia nervosa (DSM-IV) patients aged 11-17 consecutively admitted to an Eating Disorders Unit. An evaluation with laboratory data, psychopathological scales, magnetic resonance spectroscopy ((1)H MRS) and a neuropsychological battery was carried out at admission and after 7 months' follow-up and weight recovery. Psychopathological and neuropsychological and MRS examinations were also performed in 12 control subjects. In the MRS study at the frontal gray matter, the anorexic group had a significantly lower N-acetyl-aspartate (NAA) (p = .002), glutamate/glutamine (Glx) (p = .010) and myo-Inositol (mI) (p = .022) than the control group. The NAA correlated positive and significantly with triiodothyronin (Rho = .64) and the estimate level of intelligence measured with the vocabulary subtest of the WISC-R (Rho=.64). There were also positive correlations with body mass index (Rho = .47) and with attention measured with the coding subtest of the WISC-R (Rho=.51) and negative with loss of weight (Rho = -.51) but they were not statistically significant. At follow-up, there was an increase in body mass index (p=.002), triiodothyronin (p = .005), and insulin-like growth factor 1 (p = .017) and a decrease in cortisol (p = .005). In the MRS a significant increase (p = .013) in NAA was observed. The conclusion would be that NAA, Glx and mI are low in the frontal gray matter of adolescents with anorexia nervosa and specially NAA correlates with some nutritional and cognitive parameters. These alterations seem to be reversible in young patients. PMID- 17112541 TI - Is it safe to walk in the Sunbelt? Geographic variation among pedestrian fatalities in the United States, 1999-2003. AB - INTRODUCTION: Previous work using data from the 1980s showed higher rates of pedestrian mortality in the southern United States. METHODS: This study was a descriptive analysis of state-specific mortality information from the National Center for Health Statistics for 1999-2002 and the National Highway Traffic Safety Administration for 2003. RESULTS: Highest rates were in the southern rim ("Sunbelt") states for the U.S. population and for the non-Hispanic white population. Rural rates in the highest quartile were 2.1 (95% CI 1.8 to 2.6) times those in the lowest quartile. Urban rates in the highest quartile were 2.2 (95% CI 1.9 to 2.5) times those in the lowest quartile. Posted speed limits at crash sites were 2.6 (95% CI 2.0 to 3.4) times more likely to be > or = 35 mph (48.3 km/h) in the highest quartile than in the lowest quartile. Pedestrians killed in the highest quartile were 1.9 (95% CI 1.2 to 3.1) times more likely to have blood alcohol concentrations > or = 0.25 g/dL than pedestrians in the lowest quartile. CONCLUSIONS: The highest pedestrian fatality rates concentrate in Sunbelt states experiencing rapid population growth in the past 50 years. This pattern may result from at least three features of these states: (a) a high percentage of urban vehicle miles traveled; (b) urban sprawl; and (c) a high prevalence of alcohol use - especially heavy use - among Sunbelt pedestrians. PMID- 17112542 TI - Ischemia-reperfusion injury in rat skeletal muscle is attenuated by zinc aspartate. AB - BACKGROUND: Oxygen-derived free radical-induced cell injury has been suggested to have a pivotal role in the etiology of ischemia-reperfusion injury. Thus, several lines of evidence indicate that antioxidant agents may be useful therapeutics in this condition. In this regard, the effect of zinc aspartate on ischemia reperfusion injury was investigated in skeletal muscle. MATERIALS AND METHODS: Tourniquet ischemia-reperfusion injury method was applied to Sprague-Dawley rats. Experimental groups were as follows: 1) sham control, 2) rats received zinc aspartate, 3) rats received hind limb tourniquet operation (left side), and 4) rats received hind limb tourniquet operation and zinc aspartate. Viability of muscle was evaluated by triphenyltetrazolium chloride dye method by using a spectrophotometer. Malondialdehyde, superoxide dismutase, catalase, glutathione, and glutathione peroxidase were measured in muscle, heart, lung, and blood via a spectrophotometer. RESULTS: The viabilities of ischemic limbs, percentage of the contralateral control muscle, in group 1, 2, 3, and 4 were 114 +/- 12%, 87% +/- 5%, 20% +/- 2%, and 95 +/- 10%, respectively. In muscle, increased malondialdehyde and decreased superoxide dismutase, catalase, and glutathione levels in group 3 were normalized by zinc aspartate in both left and right limbs. While malondialdehyde levels in heart and blood increased in group 3, the levels of superoxide dismutase, catalase, glutathione, and glutathione peroxidase were lower in group 3 than those in group 1. All these alterations were prevented by zinc aspartate. Malondialdehyde level of lung in group 3 was significantly higher than group 1 and 2. However, this augmentation was halted by zinc aspartate. The decrease in superoxide dismutase levels in group 3 was statistically reversed by the administration of zinc aspartate. CONCLUSION: Zinc aspartate seems to be an effective treatment option against ischemia-reperfusion injury. PMID- 17112543 TI - Histochemical alterations in one lung ventilation. AB - BACKGROUND: One lung ventilation is a commonly performed surgical procedure. Although there have been several reports showing that one-lung ventilation can cause pathophysiological alterations such as pulmonary hypoxic vasoconstriction and intrapulmonary shunting, there have been virtually no reports on the effects of one-lung ventilation on lung histology. MATERIALS AND METHODS: Yorkshire pigs (11-17 kg) were anesthetized, a tracheotomy performed and a tracheal tube inserted. The chest was opened and one lung ventilation (OLV), was induced by clamping of the right main bronchus. OLV was continued for 60 min before the clamp was removed and two lung ventilation (TLV) started. TLV was continued for 30 to 60 min. Blood and lung biopsies were taken immediately before OLV, 30 min and 60 min of OLV and after restoration of TLV. RESULTS: Histological analyses revealed that the non-ventilated lung was totally collapsed during OLV. On reventilation, there was clear evidence of vascular congestion and alveolar wall thickening at 30 min after TLV. At 60 min of TLV, there was still vascular congestion. Serum nitrite levels (as an index of nitric oxide production) showed steady decline over the course of the experimental period, reaching a significantly low level on reventilation (compared with baseline levels before OLV). Lung MPO activity (marker of neutrophil sequestration) and serum TNFalpha levels were not raised during the entire experimental period. CONCLUSIONS: These results suggest that there was lung vascular injury after OLV, which was associated with reduced levels of nitric oxide production and not associated with an inflammatory response. PMID- 17112544 TI - Evolution and invasion dynamics of multiple infections with Wolbachia investigated using matrix based models. AB - Endosymbiotic bacteria are often transmitted vertically from one host generation to the next via oocytes cytoplasm. The generally small number of colonizing bacteria in the oocytes leads to a bottleneck at each generation, resulting in genetic homogenization of the symbiotic population. Nevertheless, in many of the species infected by Wolbachia (maternally transmitted bacteria), individuals do sometimes simultaneously harbor several bacterial strains, owing to the fact that Wolbachia induces cytoplasmic incompatibility (CI) that maintains multiple infections. CI occurs in crosses in which the male is infected by at least one Wolbachia strain that the female lacks, and consequently it favors individuals with the greatest symbiotic diversity. CI results in death of offspring in diploid species. In haplodiploid individuals, unfertilized eggs hatch normally into males and fertilized ones, which would lead to females, either die (female mortality type: FM) or develop into males (male development type: MD). Until now, only one theoretical study, restricted to diploid species, has investigated the associations where multiple CI-inducing Wolbachia co-exist, and explored the conditions under which multiple infections can spread. The consequences of double infections on Wolbachia maintenance in host populations, and the selective pressures to which it is subjected have not yet been analysed. Here, we have re written a model previously developed for single infection in matrix form, which allows easy extension to multiple infections and introduction of mutant strains. We show that (i) the CI type has a strong influence on invasiveness and maintenance of multiple infections; (ii) double infection lowers the invasion threshold of less competitive strains that hitch-hike with their companion strain; (iii) when multiple infections occur, as in single infections, the strains selected are those which maximize the production of infected offspring; and (iv) for the MD CI type, invasion of mutant strains can carry the whole infection to extinction. PMID- 17112545 TI - Novel method for assessing the en route survivorship of biofouling organisms on various vessel types. PMID- 17112546 TI - Aging: damage accumulation versus increasing mortality rate. AB - If aging is understood as some process of damage accumulation, it does not necessarily lead to increasing mortality rate. Within the framework of a suggested generalization of the Strehler-Mildwan (1960) [B.L. Strehler, A.S. Mildvan (1960). General theory of mortality and aging, Science, 132, 14] model, we show that even for models with monotonically increasing degradation, the mortality rate can still decrease. The decline in vitality and functions, as manifestation of aging, is modeled by the monotonically decreasing quality of life function. Using this function, the initial lifetime random variable with ultimately decreasing mortality rate is 'weighted' to result in a new random variable, which is already characterized by the increasing mortality rate. PMID- 17112547 TI - Modeling relapse in infectious diseases. AB - An integro-differential equation is proposed to model a general relapse phenomenon in infectious diseases including herpes. The basic reproduction number R(0) for the model is identified and the threshold property of R(0) established. For the case of a constant relapse period (giving a delay differential equation), this is achieved by conducting a linear stability analysis of the model, and employing the Lyapunov-Razumikhin technique and monotone dynamical systems theory for global results. Numerical simulations, with parameters relevant for herpes, are presented to complement the theoretical results, and no evidence of sustained oscillatory solutions is found. PMID- 17112548 TI - Study of the restitution of action potential duration using the artificial neural network. AB - It is widely accepted that the APD (action potential duration) restitution plays a key role in the initializing and maintaining of the reentry arrhythmias. The Luo-Rudy II models paced with different protocols showed that the current APD had a complex relation with the previous APDs and diastole intervals (DIs). This relation could not be accurately described by a single exponential function. We used an artificial neural network to formularize this relation. The results suggested that back-propagation (BP) network could predict the current APD from the information of the first three previous beats. This would help provide a target for potential anti-arrhythmic therapies. PMID- 17112549 TI - Bilateral processing in chemical synapses with electrical 'ephaptic' feedback: a theoretical model. AB - I have developed a detailed biophysical model of the chemical synapse which hosts voltage-dependent presynaptic ion channels and takes into account the capacitance of synaptic membranes. I find that at synapses with a relatively large cleft resistance (e.g., mossy fiber or giant calyx synapse) the rising postsynaptic current could activate, within the synaptic cleft, electrochemical phenomena that induce rapid widening of the presynaptic action potential (AP). This mechanism could boost fast Ca(2+) entry into the terminal thus increasing the probability of subsequent synaptic releases. The predicted difference in the AP waveforms generated inside and outside the synapse can explain the previously unexplained fast capacitance transient recorded in the postsynaptic cell at the giant calyx synapse. I propose therefore the mechanism of positive ephaptic feedback that acts between the postsynaptic and presynaptic cell contributing to the basal synaptic transmission at large central synapses. This mechanism could also explain the supralinear voltage dependence of EPSCs recorded at hyperpolarizing membrane potentials in low extracellular calcium concentration. PMID- 17112550 TI - Fishery policy when considering the future opportunity of harvesting. AB - Free access to a common pool of resource in a country may lead to over exploitation and sacrifice future opportunities of harvesting. As such, the protection of a common fishery resource is worth investigating. In this paper we develop a two-period model and a multi-period model to analyze the optimal inter temporal utilization of a finite resource of stock and propose to impose a tax on the harvest rate as an efficient mechanism with an aim at economic sustainability by incorporating the future opportunity of harvesting into the models as a major component of social objectives. The sensitivity analysis of the two-period model shows that (1) labor inputs for harvesting in Period 1 should be reduced, the biomass of fishery stock will increase, but the harvesting in Period 2 should be amplified and the biomass of fishery stock in Period 2 will not be affected if the current generation owns a higher valuation on the future opportunity of harvesting; (2) a higher internal regeneration rate leads to higher harvesting in each period and a higher level of fishery stock in Period 1, but an uncertain level of fishery stock in Period 2; (3) with a higher discount rate the harvesting in Period 1 should increase, but the harvesting in Period 2 should fall and the level of fishery stock in each period will be reduced; (4) a higher fish price in Period 1 leads to higher harvesting in Period 1, but reduced harvesting in Period 2. As a consequence, the level of fishery stock in each period will be reduced; (5) the effect of a change in fish prices in Period 2 on the harvesting and the level of fishery stock in Period 1 is uncertain, but the change in fish prices in Period 2 gives a positive effect on harvesting in Period 2 and a negative effect on the level of fishery stock in Period 2; (6) higher labor wages in Period 1 lead to lower harvesting, but a higher level of fishery stock in Period 1. This encourages an increase in harvesting in Period 2 and leads to a higher level of fishery stock in Period 2; and (7) a change of the labor wage in Period 2 affects the harvesting and the level of fishery stock in Period 1 indecisively, but it gives negative effects on the harvesting in Period 2 and positive effects on the level of fishery stock in Period 2. PMID- 17112551 TI - Food deprivation and the role of estradiol in mediating sexual behaviors in meadow voles. AB - Female mammals are particularly sensitive to changes in food availability. The mechanisms that affect sexual behavior and food intake are closely related to one another; chief among the mechanisms that control sexual behaviors in females is estradiol. In order to understand how food deprivation results in inhibition of sexual behavior (attractivity, proceptivity, and receptivity), we measured the effects of food deprivation on circulating concentrations of estradiol. We also determined whether estradiol treatment was sufficient to restore sexual behaviors in food-deprived female meadow voles. We found that estradiol titers of food deprived female voles are significantly lower than those of ad lib-fed female voles. Further, we found that estradiol treatment was sufficient to restore proceptivity and receptivity in food-deprived, ovariectomized female voles. However, estradiol treatment was not able to overcome the food deprivation induced inhibition of attractivity. Thus, decreases in estradiol titer of food deprived female voles may be related to the suppression of their proceptive and receptive behaviors, and may be a mechanism that allows females to avoid mating when conditions are not propitious for their survival and that of their offspring. PMID- 17112552 TI - Effect of previous taste experiences on taste neophobia in young-adult and aged rats. AB - Neophobia is an innate response that can be defined as the reluctance to consume novel-tasting substances. The differential effect of previous aversive and nonaversive taste memory on a subsequent neophobic response was studied in young adult (Experiment 1) and aged rats (Experiment 2). Surprising, a previous nonaversive taste experience eliminated the subsequent neophobic response to a solution of 1% sodium chloride (NaCl) in young-adult and aged rats. This result is interpreted as a generalization of the previous safe taste memory and the emotional responses that might be induced when a new taste is presented. However, a differential effect associated with aging was found for a previous aversive taste memory induced with a low dose of lithium chloride (0.15 M; 1% b.w.). While in young-adult rats this aversive taste memory did not change the neophobic response to an NaCl solution, in aged rats this memory potentiated the subsequent neophobic response to NaCl. This result is interpreted as an increase in the generalization of aversive taste memory and the emotional responses associated with aging. PMID- 17112553 TI - Minor betalains in fruits of Hylocereus species. AB - Betacyanins in peel and flesh of fruits of different Hylocereus species were identified by means of GC/MS, electrospray MS/MS, HPLC as well as (1)H and (13)C NMR techniques. As hitherto unknown pigments: betanidin 5-O-(2'-O-beta-D apiofuranosyl)-beta-D-glucopyranoside, betanidin 5-O-(4'-O-malonyl)-beta-D glucopyranoside and betanidin 5-O-[(5''-O-E-sinapoyl)-2'-O-beta-D-apiofuranosyl] beta-D-glucopyranoside were elucidated. The sinapoyl moiety attachment position in the structure of betacyanins was established for the first time. The peel contained a more complex pattern of betacyanins with apiofuranosyl moiety. Other recently identified pigments were also present in the samples and their (1)H or (13)C NMR spectra were recorded. In the case of phyllocactin and its 4'-isomer the migration of the malonyl group was noticed. PMID- 17112554 TI - Sustainable development: convergence of public health and natural environment agendas, nationally and locally. PMID- 17112555 TI - A survey of treatment practices and burden of lymphoedema in Togo. AB - Lymphatic filariasis, a mosquito-borne parasitic disease, can lead to lymphoedema and elephantiasis. This study describes the results of a baseline survey of a lymphoedema morbidity management programme in Togo. A convenience sample of 188 people with lymphoedema was asked about symptoms, treatment preferences and quality of life. Those with higher stage lymphoedema were more likely to have experienced an acute attack (odds ratio=1.9; P=0.002). Although only 28.2% of those surveyed reported currently using any lymphoedema treatment, 80.3% had used treatments in the past, primarily traditional products (68.1%) and scarification (38.8%). Medication was the preferred treatment for acute attacks, both currently (73.1%) and in the past (61.7%). Patients reported difficulties performing activities such as walking to the field (44%) and carrying a heavy load (63%) as a result of their lymphoedema. Patients felt avoided by their family (17%) and their community (36%). Using the Duke Anxiety-Depression scale, over 70% of patients were found to be at high risk of depression and this risk increased with lymphoedema stage (P=0.04). The survey results demonstrate the need for a morbidity management programme that will increase the use of morbidity management techniques and decrease the physical and emotional burden of this disease. PMID- 17112556 TI - Density-dependent host choice by disease vectors: epidemiological implications of the ideal free distribution. AB - The proportion of vector blood meals taken on humans (the human blood index, h) appears as a squared term in classical expressions of the basic reproduction ratio (R(0)) for vector-borne infections. Consequently, R(0) varies non-linearly with h. Estimates of h, however, constitute mere snapshots of a parameter that is predicted, from evolutionary theory, to vary with vector and host abundance. We test this prediction using a population dynamics model of river blindness assuming that, before initiation of vector control or chemotherapy, recorded measures of vector density and human infection accurately represent endemic equilibrium. We obtain values of h that satisfy the condition that the effective reproduction ratio (R(e)) must equal 1 at equilibrium. Values of h thus obtained decrease with vector density, decrease with the vector:human ratio and make R(0) respond non-linearly rather than increase linearly with vector density. We conclude that if vectors are less able to obtain human blood meals as their density increases, antivectorial measures may not lead to proportional reductions in R(0) until very low vector levels are achieved. Density dependence in the contact rate of infectious diseases transmitted by insects may be an important non-linear process with implications for their epidemiology and control. PMID- 17112557 TI - Two novel C29-5beta-sterols from the stems of Opuntia dillenii. AB - Two novel C29-5beta-sterols, opuntisterol [(24R)-24-ethyl-5beta-cholest-9-ene 6beta,12alpha-diol] (1) and opuntisteroside [(24R)-24-ethyl-6beta-[(beta-d glucopyranosyl)oxy]-5beta-cholest-9-ene-12alpha-ol] (2), together with nine known compounds, beta-sitosterol (3), taraxerol (4), friedelin (5), methyl linoleate (6), 7-oxositosterol (7), 6beta-hydroxystigmast-4-ene-3-one (8), daucosterol (9), methyl eucomate (10) and eucomic acid (11), were isolated from the stems of Opuntia dillenii collected in Guizhou Province, China. Their structures were elucidated mainly by spectroscopic analysis. The absolute configuration of 1 were deduced from comparative 1H NMR data of the (S)- and (R)-methoxyphenyl acetate derivatives. Compounds 6-8, 10 and 11 were isolated from O. dillenii for the first time. PMID- 17112558 TI - In vitro toxicity of silica nanoparticles in human lung cancer cells. AB - The cytotoxicity of 15-nm and 46-nm silica nanoparticles was investigated by using crystalline silica (Min-U-Sil 5) as a positive control in cultured human bronchoalveolar carcinoma-derived cells. Exposure to 15-nm or 46-nm SiO(2) nanoparticles for 48 h at dosage levels between 10 and 100 microg/ml decreased cell viability in a dose-dependent manner. Both SiO(2) nanoparticles were more cytotoxic than Min-U-Sil 5; however, the cytotoxicities of 15-nm and 46-nm silica nanoparticles were not significantly different. The 15-nm SiO(2) nanoparticles were used to determine time-dependent cytotoxicity and oxidative stress responses. Cell viability decreased significantly as a function of both nanoparticle dosage (10-100 microg/ml) and exposure time (24 h, 48 h, and 72 h). Indicators of oxidative stress and cytotoxicity, including total reactive oxygen species (ROS), glutathione, malondialdehyde, and lactate dehydrogenase, were quantitatively assessed. Exposure to SiO(2) nanoparticles increased ROS levels and reduced glutathione levels. The increased production of malondialdehyde and lactate dehydrogenase release from the cells indicated lipid peroxidation and membrane damage. In summary, exposure to SiO(2) nanoparticles results in a dose dependent cytotoxicity in cultural human bronchoalveolar carcinoma-derived cells that is closely correlated to increased oxidative stress. PMID- 17112559 TI - Recent advances in evaluation of oxime efficacy in nerve agent poisoning by in vitro analysis. AB - The availability of highly toxic organophosphorus (OP) warfare agents (nerve agents) underlines the necessity for an effective medical treatment. Acute OP toxicity is primarily caused by inhibition of acetylcholinesterase (AChE). Reactivators (oximes) of inhibited AChE are a mainstay of treatment, however, the commercially available compounds, obidoxime and pralidoxime, are considered to be rather ineffective against various nerve agents, e.g. soman and cyclosarin. This led to the synthesis and investigation of numerous oximes in the past decades. Reactivation of OP-inhibited AChE is considered to be the most important reaction of oximes. Clinical data from studies with pesticide-poisoned patients support the assumption that the various reactions between AChE, OP and oxime, i.e. inhibition, reactivation and aging, can be investigated in vitro with human AChE. In contrast to animal experiments such in vitro studies with human tissue enable the evaluation of oxime efficacy without being affected by species differences. In the past few years numerous in vitro studies were performed by different groups with a large number of oximes and methods were developed for extrapolating in vitro data to different scenarios of human nerve agent poisoning. The present status in the evaluation of new oximes as antidotes against nerve agent poisoning will be discussed. PMID- 17112560 TI - Developmental toxicity of 4-ring polycyclic aromatic hydrocarbons in zebrafish is differentially dependent on AH receptor isoforms and hepatic cytochrome P4501A metabolism. AB - Polycyclic aromatic hydrocarbons (PAHs) derived from fossil fuels are ubiquitous contaminants and occur in aquatic habitats as highly variable and complex mixtures of compounds containing 2 to 6 rings. For aquatic species, PAHs are generally accepted as acting through either of two modes of action: (1) "dioxin like" toxicity mediated by activation of the aryl hydrocarbon receptor (AHR), which controls a battery of genes involved in PAH metabolism, such as cytochrome P4501A (CYP1A) and (2) "nonpolar narcosis", in which tissue uptake is dependent solely on hydrophobicity and toxicity is mediated through non-specific partitioning into lipid bilayers. As part of a systematic analysis of mechanisms of PAH developmental toxicity in zebrafish, we show here that three tetracyclic PAHs (pyrene, chrysene, and benz[a]anthracene) activate the AHR pathway tissue specifically to induce distinct patterns of CYP1A expression. Using morpholino knockdown of ahr1a, ahr2, and cyp1a, we show that distinct embryolarval syndromes induced by exposure to two of these compounds are differentially dependent on tissue-specific activation of AHR isoforms or metabolism by CYP1A. Exposure of embryos with and without circulation (silent heart morphants) resulted in dramatically different patterns of CYP1A induction, with circulation required to deliver some compounds to internal tissues. Therefore, biological effects of PAHs cannot be predicted simply by quantitative measures of AHR activity or a compound's hydrophobicity. These results indicate that current models of PAH toxicity in fish are greatly oversimplified and that individual PAHs are pharmacologically active compounds with distinct and specific cellular targets. PMID- 17112561 TI - Tyrosine 110 in the measles virus phosphoprotein is required to block STAT1 phosphorylation. AB - The measles virus (MV) P gene encodes three proteins: P, an essential polymerase cofactor, and C and V, which have multiple functions including immune evasion. We show here that the MV P protein also contributes to immune evasion, and that tyrosine 110 is required to block nuclear translocation of the signal transducer and activator of transcription factors (STAT) after interferon type I treatment. In particular, MV P inhibits STAT1 phosphorylation. This is shown not only by transient expression but also by reverse genetic analyses based on a new functional infectious cDNA derived from a MV vaccine vial (Moraten strain). Our study also identifies a conserved sequence around P protein tyrosine 110 as a candidate interaction site with a cellular protein. PMID- 17112562 TI - Dichoptic motion perception limited to depth of fixation? AB - When counterphase spatio-temporal flicker is presented to the left and right eye continuous directional motion can be perceived. Here, we investigate whether this type of dichoptic motion can be observed at different depth planes. Four observers indicated direction of motion for dichoptic motion stimuli, presented in a context containing crossed and uncrossed disparity information in different conditions. Our results show that despite the presence of disparity cues in the stimulus, discrimination of motion direction remained maximal at interocular phase offsets that correspond to binocular motion perception at zero disparity. This constraint brings into question perception of dichoptic motion as the result of an early binocular motion system. We compared our results with predictions of a computational stereo-motion model [Qian, N. (1994). Computing stereo disparity and motion with known binocular cell properties. Neural Computations, 6, 390-404; Qian, N., & Andersen, R. A. (1997). A physiological model for motion-stereo integration and a unified explanation of Pulfrich-like phenomena. Vision Research, 37, 1683-1698]. In contrast to our empirical results, simulations of cell activation in this hybrid energy model predict maximal activation at non zero disparities. It is concluded that perception of dichoptic motion is a by product of early interocular combination at low contrasts rather than the result of a dedicated stereo-motion system. PMID- 17112563 TI - Spatial grouping in human vision: temporal structure trumps temporal synchrony. AB - Temporal information promotes visual grouping of local image features into global spatial form. However, experiments demonstrating time-based grouping typically confound two potential sources of information: temporal synchrony (precise timing of changes) and temporal structure (pattern of changes over time). Here, we show that observers prefer temporal structure for determining perceptual organization. That is, human vision groups elements that change according to the same global pattern, even if the changes themselves are not synchronous. This finding prompts an important, testable prediction concerning the neural mechanisms of binding: patterns of neural spiking over time may be more important than absolute spike synchrony. PMID- 17112564 TI - Daphnia magna and ecotoxicogenomics: gene expression profiles of the anti ecdysteroidal fungicide fenarimol using energy-, molting- and life stage-related cDNA libraries. AB - In the present study, the existing life stage-specific cDNA library was extended with energy- and molting-related genes using Suppression Subtractive Hybridization PCR and a microarray for the aquatic test organism Daphnia magna was created. A gene set of 2455 fragments was produced belonging to different pathways such as carbohydrate and lipid metabolism, O2 transport and heme metabolism, immune response, embryo development, cuticula metabolism and visual perception pathways. Using this custom microarray, gene expression profiles were generated from neonates exposed to three concentrations of the anti-ecdysteroidal fungicide fenarimol (0.5, 0.75, 1 microg/ml) during 48 h and 96 h. In total, 59 non-redundant genes were differentially expressed, of which more genes were down- than up-regulated. The gene expression data indicated a main effect on molting specific pathways. At the highest concentration, a set of proteolytic enzymes - including different serine proteases and carboxypeptidases - were induced whereas different cuticula proteins were down-regulated (48 h). Moreover, effects on embryo development were demonstrated at the gene expression as well as at the organismal level. The embryo development related gene vitellogenin was differentially expressed after 96 h of exposure together with a significant increase in embryo abnormalities in the offspring. This study suggests that this Daphnia magna microarray is of great further value for the elucidation of molecular mechanisms of toxicity and for the future development of specific biomarkers for hazard characterization. PMID- 17112565 TI - QSPR-based prediction of gas/particle partitioning of polychlorinated biphenyls in the atmosphere. AB - By reviewing the existing models for describing gas/particle partitioning of semi volatile organic compounds in the atmosphere, it was assumed that gas/particle partition coefficient, expressed as Kp, may be predicted using molecular descriptors. Overall 14 molecular descriptors of each compound calculated using semi-empirical method parametric model 3 (PM3) were tested against logKp of selected PCBs to determine the best ones governing partitioning. Eight descriptors molecular weight (Mw), molecular volume (Mv), total energy (TE), electronic energy (EE), squared atom electron densities on carbon, hydrogen and chlorine atoms in a given molecule (Summation qC2, Summation qCl2, Summation qH2) and average molecular polarizability (alpha m) were found to be highly correlated with logKp compared to other molecular descriptors. Using Partial Least-Squares Regression method (PLS), two-, three- and four-descriptor QSPR models with high fitting characters were successfully developed and their robustness and predictive power were further validated by internal cross-validation and external test. Finally, the gas/particle partition coefficients of all 209 PCBs were predicted for the first time. PMID- 17112566 TI - Do sewage treatment plant discharges substantially impair fish reproduction in polluted rivers? AB - Sewage treatment plants are frequently associated with the release of xenobiotics and, consequently, with alterations of the reproductive function induced by many of these substances in aquatic organisms. In order to assess the impacts of sewage treatment plant (STP) discharges in polluted rivers, two sentinel species (gudgeon Gobio gobio and stoneloach Barbatula barbatula) were caught during their reproductive cycle upstream and downstream two STPs (STP1--Goffontaine, STP2- Wegnez). Gonadosomatic index, histological (testicular and ovarian stages, atretic follicles, intersexuality) and endocrine (sex steroids, aromatase activity, alkali-labile phosphorus) parameters were assayed. In brief, the results revealed no systematic significant differences (p<0.05) between upstream and downstream sites, whatever the STP, species or sampling period. However, stoneloach females displayed some signs of reproductive impairment and endocrine disruption downstream STP1 (reduced GSI, oocyte diameter and ALP concentrations, increased proportion of atretic follicles) and STP2 (changes in gonadal aromatase activity and plasma levels of 11-KT and T). Few significant changes were observed for gudgeon males and females while there were no significant differences between upstream and downstream sites for stoneloach males. Moreover, plasma E(2) concentrations recorded in gudgeon males sampled in all sites were as high as in females and this was confirmed by high ALP levels. Besides, spermatogenesis of gudgeon males was delayed in STP1 upstream and downstream sites compared to the corresponding sites in STP2. These observations for gudgeon males do not seem related to STP discharge but to a probable estrogenicity of the river. Therefore, as shown by the results, stoneloach seemed more sensitive than gudgeon to STP discharges. In the present study, sewage treatment plant discharges do not substantially impair fish reproduction. In this respect, caution is required when generalising negative impacts of STP discharges. PMID- 17112567 TI - Efficient inhibition of intraperitoneal human ovarian cancer growth and prolonged survival by gene transfer of vesicular stomatitis virus matrix protein in nude mice. AB - OBJECTIVE: Vesicular stomatitis virus (VSV) matrix protein (MP) has been reported to be capable of inducing apoptosis in vitro in the absence of other viral components. In the present study, the antitumor effect of a recombinant plasmid encoding VSVMP on human ovarian cancer and its apoptosis-inducing efficacy in vivo were further investigated. METHODS: The recombinant plasmid DNA carrying VSVMP-cDNA (VSVMP-p) was constructed. SKOV3 ovarian cancer cells were transfected with VSVMP-p and examined for apoptosis by Hoechst 33258 staining and flow cytometric analysis. For in vivo study, intraperitoneal ovarian carcinomatosis models in nude mice were established and randomly assigned into four groups to receive six twice-weekly i.p. administrations of VSVMP-p/liposome complexes, empty plasmid/liposome complexes, liposome alone or 0.9% NaCl solution, respectively. The weight of intraperitoneal carcinomatosis and the survival were monitored. Tumor tissues were inspected for apoptosis by TUNEL and Hoechst-33258 assay. RESULTS: Plentiful apoptosis were observed in SKOV3 cells transfected with VSVMP-p. VSVMP-p reduced intraperitoneal tumor weight by about approximately 90% compared with control agents (p<0.01) and significantly prolonged the survival of tumor-bearing mice (p<0.05), with in vivo apoptosis index of 12.6+/-2.7% which was much higher than that of control groups (<4%) (p<0.05). Interestingly, this antitumor effect was accompanied by a noticeable NK cell accumulation. The treatment with VSVMP-p was devoid of any conspicuous toxicity. CONCLUSIONS: These observations suggest that VSVMP-p have strong antitumor effects by inducing apoptosis and possibly NK cell-mediated tumor resistance mechanisms, and it may be a potentially effective novel therapy against human ovarian cancer. PMID- 17112568 TI - The treatment of early stage cervical cancer: an assessment of pre-operative factors. AB - OBJECTIVE: To assess the pre-operative clinical factors of a group of early stage cervical cancer patients and correlate them to the risk for adjuvant radiotherapy using GOG 92 and 109 criteria. METHOD: A retrospective chart review of cervical cancer patients treated at the Saint Louis University Division of Gynecologic Oncology between the years 1989 and 2004 was performed. The results were compared with chi-squared testing and multivariable regression analysis. A p-value of 0.05 was considered significant. RESULTS: One hundred and thirty-one cervical cancer patients underwent exploration for radical hysterectomy during the study time period. Five patients had stage IA1 disease, 6 patients had stage IA2 disease, 98 patients had stage IB1 disease, 20 patients had stage IB2 disease and one patient had stage IIA disease. No patient with stage IA1 or IA2 disease met criteria for adjuvant radiotherapy. The patients with stage IB1 tumors who were 45 years of age or younger and had tumors up to 2 cm in diameter had a low (14%) likelihood for treatment with adjuvant radiotherapy. The patients with stage IB1 tumors who were older than 45 years of age with tumors larger than 2 cm in diameter and the patients with stage IB2 tumors both had a high likelihood for adjuvant radiotherapy (77% and 90% respectively). CONCLUSION: In our study group, the stage of cervical cancer and a combination of tumor diameter and patient age was found to stratify early stage cervical cancer patients by likelihood for adjuvant radiotherapy. PMID- 17112569 TI - Results of the national survey of borderline ovarian tumors in Spain. AB - MATERIAL AND METHODS: Retrospective multi-center analysis of women diagnosed with borderline ovarian tumor and treated between January 1990 and December 1997. A national survey was conducted, in which 457 patients from 27 centers corresponding to ten of Spain's autonomous communities were analyzed. RESULTS: Four hundred fifty-seven women with borderline ovarian tumor were analyzed. The mean age of patients was 45.5+/-16.9 years. Of these, 390 patients (85.3%) were at stage I, 8 (1.8%) were at stage II and 36 (7.9%) at stage III. A bilateral tumor was observed in 63 women (13.8%). The mean tumor size was 14.2 cm and in 88 cases (19.3%) the tumor was on the surface of the ovary. Microinvasion was observed in 25 (5.5%) cases, and 29 women (6.3%) showed a micropapillary pattern. Study of the factors related to the appearance of peritoneal implants revealed positive tumor markers (OR 15.02: 1.9-32.9) and a tumor on the ovarian surface (OR 8.0: 1.8-127) to be independent risk factors. With respect to recurrence, the presence of peritoneal implants at the time of initial surgery (OR 3.4: 1.1-10.4) and signs of microinvasion in the anatomicopathological study (OR 5.5: 1.5-17.8) were found to be independent risk factors. The overall survival rate in our series was 97% with a mean follow-up of 88.3 months. The survival rate by stage was 97% for stage I, 100% for stage II and 97% for stage III. CONCLUSIONS: Although borderline ovarian tumors have an excellent prognosis, they are not exempt from a risk of recurrence. Characterization of patients with borderline ovarian tumor is essential in order to prevent their evolution. Likewise, the taking on board of risk factors will enable more selective treatments to be offered in each case. PMID- 17112570 TI - Electroencephalographic and convulsant effects of the delta opioid agonist SNC80 in rhesus monkeys. AB - Non-peptidic delta opioid receptor agonists are being evaluated for a wide range of clinical applications; however, the clinical utility of piperazinyl benzamide delta agonists such as SNC80 may be limited by convulsant activity. The purpose of the present study was to evaluate the electroencephalographic and convulsant activity produced by a high dose of 10 mg/kg SNC80 IM in rhesus monkeys. EEG and behavioral activity were examined in four adult male rhesus monkeys after IM administration of SNC80. Monkeys were seated in a standard primate restraint chair, and EEG activity was recorded using an array of 16 needle electrodes implanted subcutaneously in the scalp in a bipolar (scalp-to-scalp) montage in a longitudinal direction, with bilateral frontal, central, temporal, and occipital leads. Behavior was recorded using video monitoring equipment. Initially, all monkeys were tested with 10 mg/kg SNC80, which is a relatively high dose 3-10 fold greater than doses necessary to produce a variety of other behavioral effects. Behavioral convulsions and EEG seizures were observed in one of the four monkeys. In this monkey, neither behavioral convulsions nor EEG seizures were observed when a lower dose of 3.2 mg/kg was administered nine weeks later or when the same dose of 10 mg/kg SNC80 was administered one year later. These results suggest that IM administration of SNC80 is less potent in producing convulsant effects than in producing other, potentially useful behavioral effects (e.g. antinociception) in rhesus monkeys. PMID- 17112571 TI - Intravenous methamphetamine self-administration in rats: effects of intravenous or intraperitoneal MDMA co-administration. AB - The combined use of 3,4-methylenedioxymethamphetamine (MDMA, 'Ecstasy') with methamphetamine (METH) by recreational drug users is of particular concern due to their similar pharmacological and toxic profiles. In the current study we sought to elucidate why combining these particular drugs is such a popular choice among party-drug users. This was investigated through characterisation of the possible interactive effects of MDMA on METH intravenous self-administration. The first experiment involved characterisation of the METH dose-response curve for intravenous self-administration. Male Hooded-Wistar rats were trained to self administer intravenous METH (0.01-0.3 mg/kg/infusion) and an inverted-U dose response curve was obtained. In Experiment 2, a second squad of rats self administered 0.01, 0.03 or 0.1 mg/kg/infusion METH and had small amounts of MDMA (0.001-0.03 mg/kg) then introduced into the infusion solution. Addition of MDMA to the METH infusion solution resulted in a dose independent reduction in responding. In Experiment 3, a third squad of rats was treated 20 min pre-session with an intraperitoneal injection of saline, 1.25 or 2.5 mg/kg of MDMA or METH to evaluate whether the reduction in responding evident in Experiment 2 was due to an MDMA-induced decrease in locomotor activity. Pre-treatment with intraperitoneal MDMA or METH had no effect on METH self-administration nor activity. We hypothesise that the reduction in METH self-administration caused by MDMA may reflect inhibitory effects of MDMA-induced 5-HT release on dopaminergic mechanisms. PMID- 17112572 TI - alpha-Ethyltryptamine (alpha-ET) as a discriminative stimulus in rats. AB - alpha-Ethyltryptamine (etryptamine, alpha-ET) is a drug of abuse that first appeared on the clandestine market in the mid-1980s. Its pharmacological actions are poorly understood. In this investigation, it is reported for the first time that alpha-ET serves as a training drug in drug discrimination studies. Male Sprague-Dawley rats were trained to discriminate (30-min pretreatment time) 2.5 mg/kg of alpha-ET (ED(50)=1.3 mg/kg) from saline vehicle using a standard two lever operant paradigm and a VI-15s schedule of reinforcement for appetitive reward. Once established, the alpha-ET stimulus was shown to have an onset to action of 30 min and a duration of effect of at least 4 h. In tests of stimulus generalization (substitution), the alpha-ET stimulus generalized to S(-)alpha-ET (ED(50)=1.6 mg/kg) and R(+)alpha-ET (ED(50)=1.3 mg/kg). Tests of stimulus generalization were also conducted with prototypical phenylisopropylamines: (+)amphetamine, 1-(2,5-dimethoxy-4-methylphenyl)-2-aminopropane (DOM), and N methyl-1-(4-methoxyphenyl)-2-aminopropane (PMMA). The alpha-ET stimulus generalized to DOM (ED(50)=0.4 mg/kg) and PMMA (ED(50)=0.7 mg/kg), but only partially generalized (ca. 40% maximal drug-appropriate responding) to (+)amphetamine. The results suggest that alpha-ET produces a complex stimulus. PMID- 17112573 TI - Involvement of hypothalamic pituitary adrenal axis on the nifedipine-induced antinociception and tolerance in rats. AB - Nifedipine, a calcium channel blocker, can modulate the nociceptive threshold. However, the underlying mechanism, especially the role of HPA axis, on this effect has still not been elucidated. In the present study we investigated the analgesic effect of nifedipine in intact and adrenalectomized (ADX) male rats and we also measured the effect of nifedipine on HPA function. The Tail-Flick test was used to assess the nociceptive threshold before and 15, 30, 60, 90, and 120 min after drug administration. Corticosterone level was measured by radioimmunoassay as a marker of HPA function. Our results showed that in intact and sham operated animals, administration of 10 mg/kg nifedipine induces an antinociceptive effect. But at the dosage of 2 and 5 mg/kg animals do not exhibit this effect. With repeated injections, its analgesic effect was decreased, a phenomenon prevented by adrenalectomy. Acute administration of nifedipine produced significant decrease in plasma corticosterone level. In ADX animals, had a potent antinociceptive effect nifedipine at high dosage (10 mg/kg) as well as at lower dosage (5 mg/kg) that reversed with corticosterone replacement. In conclusion, the results of our study show that the elimination of HPA function through adrenalectomy potentiates the antinociceptive effect of nifedipine and attenuates its analgesic tolerance. Both effects are reversed by corticosterone replacement. PMID- 17112574 TI - Comparative effects of dextromethorphan and dextrorphan on nicotine discrimination in rats. AB - While the role of dextrorphan and dextromethorphan as N-methyl-d-aspartate (NMDA) receptor antagonists has received considerable research attention, their effects on nicotinic acetylcholine receptors (nAChR) has been less well characterized. Recent in vitro and in vivo research has suggested that these drugs noncompetitively block alpha3beta4*, alpha4beta2, and alpha7 nAChR subtypes and antagonize nicotine's antinociceptive and reinforcing effects. Both drugs were most potent at blocking alpha3beta4* AChR. This study investigated the effects of dextrorphan and dextromethorphan on nicotine's discriminative stimulus effects. Three groups of rats were trained in a two-lever drug discrimination procedure to discriminate 0.4 mg/kg s.c. nicotine from saline. Nicotine dose-dependently substituted for itself in all three groups. In contrast, when dextrorphan (group 1) or dextromethorphan (group 2) were injected i.p., neither substitution for nor antagonism of nicotine was observed for either drug. Since i.p. administration allows substantial metabolism of dextromethorphan to its parent compound dextrorphan, the two drugs were also tested following s.c. administration (group 3). Discrimination results were similar across both routes of administration, in that neither substitution nor antagonism occurred, however, s.c. administration reduced response rates to a much greater extent than did i.p. administration. Previous work suggests that beta2 subunits are crucial for mediation of nicotine's discriminative stimulus effects and may play a role in its reinforcing effects, albeit other research suggests a role for alpha3beta4* nicotinic receptors in the latter. Our results suggest that alpha3beta4* nicotinic receptors do not play a major role in nicotine's discriminative stimulus effects. Further, they suggest that the role of cholinergic mediation of the behavioral effects of dextrorphan and dextromethorphan related to the abuse properties of nicotine may be minimal. PMID- 17112575 TI - Some rewarding effects of androgens may be mediated by actions of its 5alpha reduced metabolite 3alpha-androstanediol. AB - The abuse of anabolic-androgenic steroids (AS) is a growing problem; however, the effects and mechanisms underlying their addictive effects are not well understood. Research findings regarding androgen abuse in people and hedonic effects of androgens in laboratory rats are reviewed. Androgens, like other steroids, can have traditional actions via cognate intracellular steroid receptors, as well as other substrates. Our recent results indicate that testosterone (T) metabolites may have actions in part via gamma-aminobutyric acid (GABA)(A)/benzodiazepine receptor complexes (GBRs) and/or dopaminergic neurons in the nucleus accumbens, to mediate T's positive hedonic states. This may provide the basis for positive reinforcing effects of androgen seeking and use behavior. Following a comprehensive review of the background literature, our findings are presented that have explored the extent to which metabolites of T mediate euphorogenic effects of androgens by acting in the nucleus accumbens. Then results regarding whether GBRs are necessary substrates for androgens' positive hedonic effects are discussed. Lastly, research that addresses if dopaminergic neurons in the nucleus accumbens are necessary for these effects of androgens are discussed. This review provides a comprehensive examination of the hedonic properties and abuse/addiction potential of androgens and the putative mechanisms underlying these effects. PMID- 17112576 TI - Resveratrol improves mitochondrial function and protects against metabolic disease by activating SIRT1 and PGC-1alpha. AB - Diminished mitochondrial oxidative phosphorylation and aerobic capacity are associated with reduced longevity. We tested whether resveratrol (RSV), which is known to extend lifespan, impacts mitochondrial function and metabolic homeostasis. Treatment of mice with RSV significantly increased their aerobic capacity, as evidenced by their increased running time and consumption of oxygen in muscle fibers. RSV's effects were associated with an induction of genes for oxidative phosphorylation and mitochondrial biogenesis and were largely explained by an RSV-mediated decrease in PGC-1alpha acetylation and an increase in PGC 1alpha activity. This mechanism is consistent with RSV being a known activator of the protein deacetylase, SIRT1, and by the lack of effect of RSV in SIRT1(-/-) MEFs. Importantly, RSV treatment protected mice against diet-induced-obesity and insulin resistance. These pharmacological effects of RSV combined with the association of three Sirt1 SNPs and energy homeostasis in Finnish subjects implicates SIRT1 as a key regulator of energy and metabolic homeostasis. PMID- 17112577 TI - Protocols for synchronizing estrus and ovulation in buffalo (Bubalus bubalis): a review. AB - Poor estrus expression and a prolonged intercalving interval compromise the reproductive efficiency of female buffaloes. These limitations are exacerbated during the hot season, when fertility decreases dramatically. Pregnancy rate decrease further because difficulties in detecting estrus. To improve reproductive efficiency, several protocols of estrus and ovulation synchronization have been developed. These procedures are based on manipulating the CL, either to induce premature luteolysis using prostaglandins or to prolong the luteal phase using progestagens. However, it has recently emerged that a more precise manipulation of follicular development may be needed to achieve better synchrony of ovulation and improve fertility. Researchers have therefore turned their attention to evaluating programs in which hormones such as GnRH, FSH, LH, eCG, hCG, prostaglandins, progesterone and estradiol are administered. This review considers the impacts of estrus and ovulation synchronization protocols on fertility in the buffalo. In general, it may be stated that buffaloes respond well to the exogenous administration of hormones, and artificial insemination is possible at a pre-established time after synchronizing ovulation. Most combined hormone protocols give satisfactory pregnancy rates, comparable to those achieved in animals inseminated at natural estrus. PMID- 17112578 TI - Strengthening of a Pd-free high gold dental alloy for porcelain bonding by a pre firing heat treatment. AB - OBJECTIVES: The purpose of the present investigation was to report a Pd-free high gold alloy for porcelain bonding based on the ternary system of Au-Pt-Zn with a nominal composition of 86Au-11Pt-1.5Zn-0.5In-0.7Rh-0.2Fe-0.1Ir (wt.%). Emphasis was put on the effect of a pre-firing heat treatment on the mechanical properties of the alloy. METHODS: The strengthening effect of the pre-firing heat treatment was investigated by means of hardness measurement, tensile testing, X-ray diffraction, optical microscopy and scanning electron microscopy. RESULTS: Experimental results showed that both the hardness and tensile strength of the alloy can be significantly improved after heat treatment at the temperature of 980 degrees C for 15 min. The strengthening could be attributed to homogenization of microstructure and alloying elements and precipitation of new fine particles. SIGNIFICANCE: The cast framework of the present new Pd-free alloy could be heat treated before actual firing, and this would improve the processing properties of the alloy during firing. PMID- 17112579 TI - Characterization and surface treatment effects on topography of a glass infiltrated alumina/zirconia-reinforced ceramic. AB - OBJECTIVE: Characterize the microstructure, composition and some physical properties of a glass-infiltrated alumina/zirconia-reinforced ceramic (IZ) and the effect of surface treatment on topography. METHODS: IZ ceramic specimens were fabricated according to ISO6872 instructions and polished through 1 microm alumina abrasive. Quantitative and qualitative analyses were performed using scanning electron microscopy (SEM), backscattered imaging (BSI), electron dispersive spectroscopy (EDS) and stereology. The elastic modulus (E) and Poisson's ratio (nu) were determined using ultrasonic waves, and the density (rho) using a helium pycnometer. The following ceramic surface treatments were used: AP-as-polished; HF-etching with 9.5% hydrofluoric acid for 90 s; SB sandblasting with 25 microm aluminum oxide particles for 15s and SC-blasting with 30 microm aluminum oxide particles modified by silica (silica coating) for 15s. An optical profilometer was used to examine the surface roughness (Ra) and SEM EDS were used to measure the amount of silica after all treatments. RESULTS: The IZ mean property values were as follows: rho=4.45+/-0.01 g/cm(3); nu=0.26 and E=245 GPa. Mean Ra values were similar for AP- and HF-treated IZ but significantly increased after either SC or SB treatment (p0.50 mg/dl, elevated CRP group, N=5) scored significantly higher than those with CRP levels in the normal range (CRP220,000 individuals which had been covered by an insurance plan between January 1, 1995 and December 31, 2002. Out of these, 1235 CAD cases were identified and matched to postal-code areas based upon the address of the owner. Environmental risk factors we considered included averages of long-term annual rainfall, and of January and July temperatures. Initial visualization of the incidence rate of CAD (cases/dog-years at risk) expressed as empirical Bayes smoothed spatial rates indicated geographic variation. Moran's I, adjusted for population at risk, revealed significant global clustering. Both the spatial scan statistic and a local indicator of spatial autocorrelation revealed a higher incidence of CAD in the major cities. In a Poisson-regression model (with a spatial covariance structure), the incidence of CAD increased with increasing human population density, increasing average annual rainfall, living in the southern half of Sweden, and having a veterinary dermatologist in the county. PMID- 17112612 TI - Impact of sedation method on the diagnosis of hip and elbow dysplasia in Swedish dogs. AB - Our objective was to investigate the effect of sedation method on the screening result for hip and elbow dysplasia. The study was based on a questionnaire survey of routines for hip and elbow screening at Swedish veterinary clinics and results of hip and elbow status, for eight breeds (Bernese Mountain Dog, Boxer, German Shepherd Dog, Golden Retriever, Labrador Retriever, Newfoundland, Rottweiler, and Saint Bernard) recorded by the Swedish Kennel Club. In total 5877 and 5406 dogs examined for hip and elbow dysplasia, respectively, from January 2002 through March 2003 were included. We used logistic regression to examine whether the type of chemical restraint used for sedation affected the screening result for hip and elbow dysplasia. In addition to sedation method, the effects of veterinary clinic, sex, breed, and age at screening were studied. The type of chemical restraint used for sedation affected the screening result for hip but not for elbow dysplasia. Acepromazine gave less than half the odds of hip dysplasia compared with medetomidine and butorphanol (the most common method), medetomidine alone or xylazine. Females had about 25% higher odds for developing hip dysplasia whereas males had almost 40% higher odds for developing elbow dysplasia. Saint Bernard, Newfoundland and German Shepherd Dog had the highest odds of developing hip dysplasia, whereas Rottweiler and Labrador Retriever had the lowest odds. Boxer had the lowest risk for elbow dysplasia, followed by Labrador Retriever. Saint Bernard and Rottweiler had the highest odds of elbow dysplasia. Increasing age increased the odds of both hip and elbow dysplasia, by about 2.5% per month. Following the results in this study, recording of the type of chemical restraint used for sedation during hip screening has now become mandatory in Sweden. This makes it possible to account for the effect of sedation method in a model for prediction of breeding values for hip dysplasia. PMID- 17112613 TI - Do health sector reforms have their intended impacts? The World Bank's Health VIII project in Gansu province, China. AB - This paper combines differences-in-differences with propensity score matching to estimate the impacts of a health reform project in China that combined supply side interventions aimed at improving the effectiveness and quality of care with demand-side measures aimed at expanding health insurance and providing financial support to the very poor. Data from household, village and facility surveys suggest the project reduced out-of-pocket spending, and the incidence of catastrophic spending and impoverishment through health expenses. Little impact is detected on the use of services, and while the evidence points to the project reducing sickness days, the evidence on health outcomes is mixed. PMID- 17112614 TI - Comparison of amplification methods for transcriptomic analyses of low abundance prokaryotic RNA sources. AB - Microarrays have established as instrumental for bacterial detection, identification, and genotyping as well as for transcriptomic studies. For gene expression analyses using limited numbers of bacteria (derived from in vivo or ex vivo origin, for example), RNA amplification is often required prior to labeling and hybridization onto microarrays. Evaluation of the fidelity of the amplification methods is crucial for the robustness and reproducibility of microarray results. We report here the first utilization of random primers and the highly processive Phi29 phage polymerase to amplify material for transcription profiling analyses. We compared two commercial amplification methods (GenomiPhi and MessageAmp kits) with direct reverse-transcription as the reference method, focusing on the robustness of mRNA quantification using either microarrays or quantitative RT-PCR. Both amplification methods using either poly A tailing followed by in vitro transcription, or direct strand displacement polymerase, showed appreciable linearity. Strand displacement technique was particularly affordable compared to in vitro transcription-based (IVT) amplification methods and consisted in a single tube reaction leading to high amplification yields. Real-time measurements using low-, medium-, and highly expressed genes revealed that this simple method provided linear amplification with equivalent results in terms of relative messenger abundance as those obtained by conventional direct reverse-transcription. PMID- 17112615 TI - Eye movements and prospective memory: what the eyes can tell us about prospective memory. AB - In this study we used eye tracking methodology in combination with multi-element displays to examine the processes underlying event-based prospective memory in a visual search paradigm. In the task individuals searched for a different target stimulus (i.e., a letter) on each trial that could be present or absent, and made prospective responses to the letters D or M. The response accuracy data revealed that target hits were more frequent than prospective hits, and that there was no difference in response time for target and prospective responses. The eye tracking data revealed that both first and total fixation durations increased from distractors (stimuli that were neither targets nor prospective cues) to targets to prospective cues when a target or prospective response was made. These measures also revealed that the presence of a target in the display served to disrupt prospective memory. In addition, prospective memory misses resulted from a combination of failures to fixate the prospective cues and failures to engage in strategic processing of the prospective cues. These data demonstrate the utility of eye tracking methods in examining the processes underlying event-based prospective memory. PMID- 17112616 TI - Baculovirus-mediated gene silencing in insect cells using intracellularly produced long double-stranded RNA. AB - Double-stranded RNA-mediated interference (RNAi) has recently emerged as a powerful reverse genetics tool to silence gene expression in multiple organisms, including plants, nematodes and insects. In this study, DNA vectors capable of promoting the synthesis of long hairpin dsRNAs in vivo from a DNA template to suppress gene expression in insect cells have been successfully constructed. The inhibition of the expression of a gene encoding enhanced green fluorescent protein (eGFP) in insect cells was demonstrated by using plasmid or baculovirus vectors. Both plasmid and baculovirus vectors were able to inhibit eGFP expression in a dose dependent manner. Complete inhibition was obtained when co transfection ratios of target plasmid to inhibition plasmid were 1:1 and 1:0.1. Eighty percent suppression was still maintained even when the ratio of eGFP plasmid to 'hairpin' plasmid was as high as 1:0.01. When the hairpin dsRNAs were encoded in a baculovirus, the suppression was about 50% when the ratio of 'target' baculovirus to 'inhibition' baculovirus reached 1:10. Therefore, the designed plasmid and baculovirus vectors are useful to induce RNAi in insect cell systems. PMID- 17112617 TI - Molecular dissection of nuclear entry-competent SV40 during infection. AB - To establish viral infection, SV40 must expose nuclear localization signals (NLSs) that are internal in the virion architecture in order to enter the nucleus via interaction with the host's nuclear import machinery, which includes importin alpha and importin beta. The time course for SV40 association with the importins in infected cells was examined. The viral DNA associated with importin alpha by 1.5h post infection, before associating with the importin beta nuclear import receptor, by 3h post infection. Only a small fraction of cell-internalized SV40 that contained viral DNA was bound by the two importins. This fraction, termed "nuclear entry-competent SV40," was slightly smaller than the virion but, importantly, was larger than the viral chromatin and contained both Vp1 and Vp3. Furthermore, the internalized viral DNA in either anti-importin or anti-Vp3 immune complexes was sensitive to DNase I, whereas the viral DNA in mature virions was resistant. All these results suggest that once SV40 enters the cytoplasm, it undergoes an architectural modification that exposes the virion's NLSs for nuclear entry. PMID- 17112618 TI - Uptake and metabolism of novel biodegradable poly (glycerol-adipate) nanoparticles in DAOY monolayer. AB - A useful route for the development of antitumour therapies is by creating improved methods for delivering therapeutic agents to tumour cells or subcellular compartments and increasing retention of drugs within target cells. In this study, we have characterized nanoparticle (NP) uptake and metabolism by DAOY cells, a human medulloblastoma cell line. NPs were formed from a novel polymer, poly (glycerol-adipate) (PGA), containing Rhodamine B Isothiocyanate (RBITC) as a fluorescent marker. It was observed that the cellular uptake of NPs depends on the incubation time and the concentration of NPs in the culture medium. The studies of retention and metabolism of NPs within cells indicated that 1) faster degradation of NPs within cells compared with that in cell culture medium in vitro; 2) a small fraction of NPs were recycled back to the outside of cell, whereas most NPs entered endosomes and lysosomes; and 3) recycled NPs were re taken up in the following 2 h incubation time. These studies thus suggested that PGA NPs could be used for localising therapeutic agents into cells, and could provide prolonged drug effects because of their long sustained release in physiological conditions and their rapid release when taken up into cells. PMID- 17112619 TI - Optimising the incorporation and release of a neurotrophic factor using conducting polypyrrole. AB - In this study, a neurotrophin delivery system based on an inherently conducting polymer (ICP) has been developed. Direct incorporation of neurotrophin-3 (NT-3) was investigated and controlled release was tested under various electrochemical conditions. The loading capacity and amount of NT-3 released from the polymer was determined using (125)I-labelled NT-3. Electrochemical stimulation of polypyrrole by pulsed voltage, pulsed current or cyclic voltammetry promoted the release of NT-3 at a greater rate than natural diffusion of NT-3. NT-3 was released from polypyrrole as an initial burst in the first 24 h followed by prolonged release over a subsequent 6 days of sampling. The amount of NT-3 incorporated into the polymer could be controlled by varying the polymerisation time, with longer growth periods incorporating more NT-3. The NT-3 release results indicated that the polymers grown for longer released a lower percentage of the incorporated NT 3 compared to the polymers grown for shorter times. Polymer-based neurotrophin delivery systems have the potential to be incorporated into future treatments for nerve injuries to prevent nerve degradation and promote nerve protection. PMID- 17112620 TI - Monocytes from type 2 diabetic patients have a pro-inflammatory profile. 1,25 Dihydroxyvitamin D(3) works as anti-inflammatory. AB - The exact factors contributing to the pathogenesis of type 2 diabetes remain elusive. Lately, it was suggested that inflammation and activation of the innate immune system could be linked to type 2 diabetes pathogenesis and also to the development of common diabetic complications, mainly atherosclerosis. The aim of this study was to investigate the role of monocytes in this sub-clinical inflammatory state and test 1,25-dihydroxyvitamin D(3), the active form of Vitamin D, as an anti-inflammatory agent. For this purpose, monocytes from type 2 diabetic patients were compared to monocytes from healthy controls and type 1 diabetic patients. The expression profile of inflammatory markers in freshly isolated and immune-stimulated monocytes was measured by quantitative real-time RT-PCR. Type 2 diabetic patients showed significantly higher expression levels of TNF-alpha, IL-6, IL-1, IL-8, COX-2, ICAM-1 and B7-1 compared to controls and type 1 diabetic patients. 1,25-Dihydroxyvitamin D(3) was able to down-regulate the expression of TNF-alpha, IL-6, IL-1, and IL-8, confirming its immunomodulatory properties. From these data we concluded that monocytes from type 2 diabetic patients have a pro-inflammatory profile. In addition, 1,25-dihydroxyvitamin D(3) was able to modulate inflammation in these monocytes. PMID- 17112621 TI - A review of human and analogue insulin trials. AB - A recent meta-analysis evaluated trials of the rapid-acting analogues insulin lispro and insulin aspart, performed before the introduction of the basal analogues, insulin glargine and insulin detemir. This article reviews the effect of rapid-acting and basal insulin analogues separately and in combination, relative to human insulin. Outcomes evaluated include HbA(1c), hypoglycaemia, postprandial glucose (PPG), and weight changes. Results from trials that matched defined criteria are presented in tables. In type 1 diabetes, compared with human insulin, the rapid-acting analogues generally reduced hypoglycaemia and postprandial glucose, whereas the basal analogues tended to reduce hypoglycaemia - particularly nocturnal hypoglycaemia. Weight gain may also be reduced with basal analogues, compared with human basal insulin. In type 2 diabetes, premix rapid-acting analogues controlled postprandial glucose better than human insulin mixes; basal analogues used as basal-only therapy reduced hypoglycaemia compared with NPH insulin; and some advantages were apparent with analogues in basal-bolus therapy. Whilst the benefits on individual metabolic and clinical outcomes appear modest, almost all studies report some advantage when using insulin analogues in type 1 and type 2 diabetes. Significant benefits, including PPG lowering with the rapid-acting analogues and the potential for reduction in cardiovascular risk, should be investigated further. PMID- 17112622 TI - Occult hepatitis B virus infection. AB - The persistence of hepatitis B virus (HBV) genomes in HBV surface antigen (HBsAg) negative individuals is termed occult HBV infection. Occult HBV status is associated in some cases with mutant viruses undetectable by HBsAg assays, but more frequently it is due to a strong suppression of viral replication and gene expression. Occult HBV infection is an entity with world-wide diffusion, although the available data of prevalence in various categories of individuals are often contrasting because of the different sensitivity and specificity of the methods used for its detection in many studies. Occult HBV may impact in several different clinical contexts, including the transmission of the infection by blood transfusion or organ transplantation and its acute reactivation when an immunosuppressive status occurs. Moreover, much evidence suggests that it can favour the progression of liver fibrosis and above all the development of hepatocellular carcinoma. PMID- 17112623 TI - In the new area of noninvasive markers of hepatocellular carcinoma. PMID- 17112624 TI - Clinical evaluation (Phase I) of a human monoclonal antibody against hepatitis C virus: safety and antiviral activity. AB - BACKGROUND/AIMS: HCV-AB68, a human monoclonal antibody against the envelope protein of hepatitis C virus (HCV), neutralizes HCV in cell-culture and in the HCV-Trimera mouse model. A Phase 1 clinical trial was designed to test safety, tolerability, and antiviral activity of HCV-AB68 in patients with chronic HCV infection. METHODS/RESULTS: Single doses of HCV-AB68, 0.25-40 mg, administered to 15 patients were well tolerated with no moderate or serious adverse events (SAEs) reported. In six patients, HCV-RNA levels transiently decreased by 2- to 100-fold immediately following infusion and rebound to baseline in 24-48 h. Multiple doses of HCV-AB68, 10-120 mg, were administered to 25 patients. Doses were given weekly for 3 weeks, then 3x a week during the fourth week, after which patients were followed for 3 months. No drug-related SAEs were reported and no specific pattern of adverse events was evident. Eight out of 25 patients had at least a 1-log reduction and 17 had at least a 0.75-log reduction in HCV-RNA levels from baseline at one or more time points following HCV-AB68 infusion. CONCLUSIONS: These data support the investigation of HCV-AB68 in the prevention of recurrent HCV-infection in patients who had received hepatic allografts for end-stage liver disease. PMID- 17112625 TI - Going towards more relevant cell culture models to study the in vitro replication of serum-derived hepatitis C virus and virus/host cell interactions? PMID- 17112626 TI - Attenuated liver progenitor (oval) cell and fibrogenic responses to the choline deficient, ethionine supplemented diet in the BALB/c inbred strain of mice. AB - BACKGROUND/AIMS: Liver regeneration following chronic injury is associated with inflammation, the proliferation of liver progenitor (oval) cells and fibrosis. Previous studies identified interferon-gamma as a key mediator of oval cell proliferation. Interferon-gamma is known to regulate Th1 cell activities during immune challenge. Therefore, we hypothesised that progenitor cell-mediated regeneration is associated with a Th1 immune response. METHODS: C57Bl/6 (normal Th1 response) and BALB/c mice (deficient in Th1 signalling) were placed on a carcinogenic diet to induce liver injury, progenitor cell proliferation and fibrosis. RESULTS: Serum transaminases and mortality were elevated in BALB/c mice fed the diet. Proliferation of liver progenitor cells was significantly attenuated in BALB/c animals. The pattern of cytokine expression and inflammation differed between strains. Liver fibrosis and hepatic stellate cell activation were significantly inhibited in BALB/c mice compared to C57Bl/6. In addition, interferon-gamma knockout mice also showed reduced fibrosis compared to wild type. These findings are in contrast to published results, in which interferon gamma is shown to be anti-fibrogenic. CONCLUSIONS: Our data demonstrate that the hepatic progenitor cell response to a CDE diet is inhibited in mice lacking Th1 immune signalling and further show that this inhibition is associated with reduced liver fibrosis. PMID- 17112627 TI - Curative bone marrow transplantation in erythropoietic protoporphyria after reversal of severe cholestasis. AB - We report the case of a middle-age patient presenting with severe progressive protoporphyric cholestasis. To halt further progression of liver disease, medical treatment was given aimed at different mechanisms possibly causing cholestasis in erythropoietic protoporphyria. Within eighty days, liver biochemistry completely normalized and liver histology markedly improved. Bone marrow transplantation was performed to prevent relapse of cholestatic liver disease by correcting the main site of protoporphyrin overproduction. Thirty-three months after cholestatic presentation and ten months after bone marrow transplantation, liver and porphyrin biochemistry remains normal. The patient is in excellent condition and photosensitivity is absent. The theoretical role of each treatment used to successfully reverse cholestasis and the role of bone marrow transplantation in erythropoietic protoporphyria are discussed. Medical treatment can resolve hepatic abnormalities in protoporphyric cholestasis. Bone marrow transplantation achieves phenotypic reversal and may offer protection from future protoporphyric liver disease. PMID- 17112628 TI - Does treatment with interferon-based therapy improve the natural history of chronic hepatitis B infection? PMID- 17112629 TI - Serum-derived hepatitis C virus infectivity in interferon regulatory factor-7 suppressed human primary hepatocytes. AB - BACKGROUND/AIMS: The development of an efficient in vitro infection system for HCV is important in order to develop new anti-HCV strategy. Only Huh7 hepatocyte cell lines were shown to be infected with JFH-1 fulminant HCV-2a strain and its chimeras. Here we aimed to establish a primary hepatocyte cell line that could be infected by HCV particles from patients' sera. METHODS: We transduced primary human hepatocytes with human telomerase reverse transcriptase together with human papilloma virus 18/E6E7 (HPV18/E6E7) genes or simian virus large T gene (SV40 T) to immortalize cells. We also established the HPV18/E6E7-immortalized hepatocytes in which interferon regulatory factor-7 was inactivated. Finally we analyzed HCV infectivity in these cells. RESULTS: Even after prolonged culture HPV18/E6E7 immortalized hepatocytes exhibited hepatocyte functions and marker expression and were more prone to HCV infection than SV40 T-immortalized hepatocytes. The susceptibility of HPV18/E6E7-immortalized hepatocytes to HCV infection was further improved, in particular, by impairing signaling through interferon regulatory factor-7. CONCLUSIONS: HPV18/E6E7-immortalized hepatocytes are useful for the analysis of HCV infection, anti-HCV innate immune response, and screening of antiviral agents with a variety of HCV strains. PMID- 17112630 TI - Leptin and liver tissue repair: do rodent models provide the answers? PMID- 17112631 TI - Simulation of biological ion channels with technology computer-aided design. AB - Computer simulations of realistic ion channel structures have always been challenging and a subject of rigorous study. Simulations based on continuum electrostatics have proven to be computationally cheap and reasonably accurate in predicting a channel's behavior. In this paper we discuss the use of a device simulator, SILVACO, to build a solid-state model for KcsA channel and study its steady-state response. SILVACO is a well-established program, typically used by electrical engineers to simulate the process flow and electrical characteristics of solid-state devices. By employing this simulation program, we have presented an alternative computing platform for performing ion channel simulations, besides the known methods of writing codes in programming languages. With the ease of varying the different parameters in the channel's vestibule and the ability of incorporating surface charges, we have shown the wide-ranging possibilities of using a device simulator for ion channel simulations. Our simulated results closely agree with the experimental data, validating our model. PMID- 17112632 TI - A Kalman filter based methodology for EEG spike enhancement. AB - In this work, we present a methodology for spike enhancement in electroencephalographic (EEG) recordings. Our approach takes advantage of the non stationarity nature of the EEG signal using a time-varying autoregressive model. The time-varying coefficients of autoregressive model are estimated using the Kalman filter. The results show considerable improvement in signal-to-noise ratio and significant reduction of the number of false positives. PMID- 17112633 TI - Improving hospital surge capacity: a new concept for emergency credentialing of volunteers. AB - In the event of a large-scale terrorist attack, natural disaster, or other public health emergency, hospitals could not absorb the thousands of victims generated by the catastrophe. Even if hospitals can increase bed capacity by 20% to 30%, as some suggest, the problem of staffing these beds remains unresolved. One possibility is to rapidly increase hospital staff by providing emergency credentialing to volunteer health care professionals. Several organizations and systems currently exist that can deliver medical providers to a stricken area. Unfortunately, all of these have serious limitations that would make it difficult for hospitals to use the health care workers provided by such entities. We propose a unique concept that will allow hospitals to rapidly expand their staff with practitioners that meet their credentialing requirements. The concept is a database created by each hospital in a community that includes credentialed physicians, nurses, behavioral health professionals, and ancillary staff. The database will be limited to physicians with full privileges and all licensed hospital employees in good standing not currently facing disciplinary issues or practice restrictions. The individual databases would then be combined and stored on a single computer system housed at the county health care agency or other mutually acceptable organization, with copies sent back to participating hospitals and the state. After a large disaster, health care workers from unaffected areas, including other states, can approach affected hospitals and volunteer their services. Practitioners listed on the database could be given privileges in their specialties for 72 hours. This process is accurate, inexpensive, efficient, sustainable, and Joint Commission on Accreditation of Healthcare Organizations compliant and permits the immediate credentialing of large numbers of medical volunteers. PMID- 17112634 TI - Trauma management outcomes associated with nonsurgeon versus surgeon trauma team leaders. AB - STUDY OBJECTIVE: We compare the effectiveness of surgeon and nonsurgeon trauma team leaders. METHODS: This retrospective study was conducted using data from a Canadian trauma registry database. Data from April 1, 1998, to March 31, 2005, from blunt and penetrating trauma patients aged 16 years or older and with trauma team activation (and without major burns) were included. Patient age, sex, trauma team leader (surgeon or nonsurgeon), mechanism of injury, Injury Severity Score, survival to 3 hours and to discharge, length of stay in the hospital, and Trauma and Injury Severity Score (TRISS) z scores were tabulated. RESULTS: Data from 807 patients were included. Because of the limited number of penetrating trauma cases, analyses focused on blunt trauma. Surgeon and nonsurgeon trauma team leader groups did not differ on injury severity, age, or sex. No difference was noted in survival to discharge (nonsurgeon 84.8%-surgeon 81.8%=3%; 95% confidence interval [CI] -3.5% to 9.5%), survival to 3 hours (nonsurgeon 96.8%-surgeon 96%=0.8%; 95% CI -2.2% to 3.8%), length of stay (median 13 days for nonsurgeon and 12 days for surgeon groups), or difference between actual and predicted survival (TRISS z scores nonsurgeon 0.64; surgeon 0.99). No trend toward group differences on any outcome variable was observed in penetrating trauma cases. CONCLUSION: No differences were found in the outcome of trauma patients treated by nonsurgeon versus surgeon trauma team leaders. These findings support a more collaborative approach to resuscitative trauma management with involvement of nonsurgeons as trauma team leaders. PMID- 17112635 TI - Intraneuronal amyloid beta and reduced brain volume in a novel APP T714I mouse model for Alzheimer's disease. AB - Transgenic mouse models of Alzheimer's disease (AD) expressing high levels of amyloid precursor protein (APP) with familial AD (FAD) mutations have proven to be extremely useful in understanding pathogenic processes of AD especially those that involve amyloidogenesis. We earlier described Austrian APP T714I pathology that leads to one of the earliest AD age-at-onsets with abundant intracellular and extracellular amyloid deposits in brain. The latter strikingly was non fibrillar diffuse amyloid, composed of N-truncated A beta 42 in absence of A beta 40. In vitro, this mutation leads to one of the highest A beta 42/A beta 40 ratios among all FAD mutations. We generated an APP T714I transgenic mouse model that despite having 10 times lower transgene than endogenous murine APP deposited intraneuronal A beta in brain by 6 months of age. Accumulations increased with age, and this was paralleled by decreased brain sizes on volumetric MRI, compared to age-matched and similar transgene-expressing APP wild-type mice, although, with these levels of transgenic expression we did not detect neuronal loss or significant memory impairment. Immunohistochemical studies revealed that the majority of the intraneuronal A beta deposits colocalized with late endosomal markers, although some A beta inclusions were also positive for lysosomal and Golgi markers. These data support earlier observations of A beta accumulation in the endosomal-lysosomal pathway and the hypothesis that intraneuronal accumulation of A beta could be an important factor in the AD pathogenesis. PMID- 17112636 TI - Donepezil markedly potentiates memantine neurotoxicity in the adult rat brain. AB - The NMDA antagonist, memantine (Namenda), and the cholinesterase inhibitor, donepezil (Aricept), are currently being used widely, either individually or in combination, for treatment of Alzheimer's disease (AD). NMDA antagonists have both neuroprotective and neurotoxic properties; the latter is augmented by drugs, such as pilocarpine, that increase cholinergic activity. Whether donepezil, by increasing cholinergic activity, might augment memantine's neurotoxic potential has not been investigated. In the present study, we determined that a dose of memantine (20mg/kg, i.p.), considered to be in the therapeutic (neuroprotective) range for rats, causes a mild neurotoxic reaction in the adult rat brain. Co administration of memantine (20 or 30 mg/kg) with donepezil (2.5-10mg/kg) markedly potentiated this neurotoxic reaction, causing neuronal injury at lower doses of memantine, and causing the toxic reaction to become disseminated and lethal to neurons throughout many brain regions. These findings raise questions about using this drug combination in AD, especially in the absence of evidence that the combination is beneficial, or that either drug arrests or reverses the disease process. PMID- 17112637 TI - Promoter polymorphisms which modulate APP expression may increase susceptibility to Alzheimer's disease. AB - Increasing evidence indicates that variants in promoter of the beta-amyloid precursor protein (APP) gene could up-regulate the APP gene expression and aggravate the amyloid beta protein (A beta) accumulation, thus contributing to the development of Alzheimer's disease (AD). In Chinese Han populations we found three polymorphisms in APP promoter: -877T/C(rs466433), -955A/G(rs364048) and 9G/C. The -877T and -955A alleles were over-represented in 209 sporadic AD (SAD) patients when compared to those in 437 healthy individuals. Furthermore, -877T/C and -955A/G were in strong linkage disequilibrium and they constructed a relatively risky -877T/-955A and a relatively protective -877C/-955G. Luciferase reporter assay indicated -877T/-955A had four times higher transcriptional activity than -877C/-955G. A more marked increase in -877T/-955A transcriptional activity was seen when under A beta(25-35) treatment. As for the -9G/C polymorphism, significant differences between the two alleles were not observed either in genetic evaluation or in functional assay. The present study provides strong evidence that APP promoter polymorphisms that significantly increase APP expression levels are associated with development of SAD. PMID- 17112638 TI - Design, parallel synthesis and SAR of novel urotensin II receptor agonists. AB - A 30-membered library of amides based on the potent urotensin II (UII) receptor agonist FL104, has been synthesized from ten different carboxylic acids and three amines. A synthetic protocol producing the amides in 47-98% yield has been developed in which the purification involved only extractions and in a few cases filtration through an ion-exchange resin. It was found that 5mg of starting material was enough to obtain reproducible results and excellent purities. Thus, the procedure is estimated to be transferable to fully automated systems. The synthesized compounds were evaluated for their UII receptor agonistic activities using a cell-based assay (R-SAT). The most active compounds were the 4 trifluoromethylcinnamic amides of 1-(4-chlorophenyl)-3-dimethylamino-propylamine and 1-(2-naphthyl)-3-dimethylamino-propylamine, both showed EC(50) values of 130 nM. PMID- 17112639 TI - Effect of substituents on diarylmethanes for antitubercular activity. AB - Aminoalkyl derivatives of diarylmethanes were prepared using Grignard, Friedel Crafts arylation and aminohydrochloride chain formation reactions. These series of compounds were evaluated against Mycobacterium tuberculosis H(37)R(v) and showed the activity in the range of 6.25-25 microg/mL. Effect of heteroaryl, anthracenyl and phenanthrene groups on diarylmethane pharmacophores for antitubercular activity is described. PMID- 17112640 TI - A review of anti-infective and anti-inflammatory chalcones. AB - Chalcones, considered as the precursors of flavonoids and isoflavonoids, are abundant in edible plants, and have also been shown to display a diverse array of pharmacological activities. The purpose of this review is to provide an overview of the pharmacological activity of synthetic and naturally occurring chalcones. This review is complementary to earlier reviews and covers more recent reports of antimicrobial activity of chalcones (antibacterial and antifungal), as well as antileishmanial, antimalarial, antiviral and anti-inflammatory activities of these compounds. PMID- 17112641 TI - Synthesis of isonicotinoylhydrazones from anacardic acid and their in vitro activity against Mycobacterium smegmatis. AB - Isonicotinoylhydrazones were synthesized from a natural product anacardic acid, a major constituent of cashew nut shell liquid. The unsaturated side chain in anacardic acid and its 5-nitro derivative were converted into C(8')-aldehydes by oxidative cleavage. C(8')-aldehydes are then coupled with isoniazid (an anti-TB drug) to obtain N-isonicotinoyl-N'-8-[(2'-carbohydroxy-3'-hydroxy) phenyl] octanal hydrazone (5) and N-isonicotinoyl-N'-8-[(2'-carbohydroxy-3'-hydroxy-6 nitro) phenyl] octanal hydrazone (6). These isonicotinoylhydrazones of anacardic aldehydes showed potent antimycobacterial activity against Mycobacterium smegmatis mc(2)155. The synergistic studies of 5 and 6 with isoniazid showed more inhibitory activities than isoniazid alone. Compounds 5 and 6 also showed activity against Mycobacterium tuberculosis H(37)Rv. PMID- 17112642 TI - New polyamine-sensitive inhibitors of the NMDA receptor: syntheses and pharmacological evaluation. AB - Derivatives of 5-(4-aminobutyl)-2-thiophene-octylamine, a potent polyamine sensitive inhibitor of the NMDA receptor, were synthesized and evaluated as inhibitors of [(3)H]MK-801 binding to rat brain membranes. Alkylations of the terminal amino groups reduced inhibitory potency; only incorporation of the amino group of the short 4-aminobutyl arm into a piperidine ring was tolerated. Substitution of the thiophene nucleus with methyl or ethyl, and its replacement by a benzene nucleus, was of minor influence. The corresponding diguanidines exhibited high potency independent of chain length, whereas their sensitivity to spermine was sharply dependent on chain length. Insertion of an amide bond into the long octylamine arm increased sensitivity to spermine and to Tris buffer. Our results indicate that spermine sensitivity of [(3)H]MK-801 binding inhibition is responsive to subtle changes in inhibitor structure and represents a promising target for pharmaceutical research. PMID- 17112643 TI - Comprehensive epitope analysis of cross-clade Gag-specific T-cell responses in individuals with early HIV-1 infection in the US epidemic. AB - To elucidate the mechanisms underlying cross-clade T-cell reactivity, we evaluated responses to Gag peptides based on clades A, B, C, and M-group sequences at the epitope level by IFN-gamma ELISpot assay in 25 subjects following primary clade B infection. T-cell reactivity to CON (consensus), COT (center of tree), and ANC (most recent common ancestor) B peptides was similar and a high level of cross-reactivity was noted to clade A, C, and M-group peptides. T-cell responses to 15 of the 16 epitopes reacted with at least 1 of the 2 heterologous peptides (A or C or both) and 7 epitopes were invariant across all 3 clades. The remaining 9 epitopes were associated with a total of 11 variant sequences, and with the exception of 1, all substitutions were outside the HLA anchor positions. We conclude that Gag-specific cross-clade T-cell responses producing IFN-gamma can be detected in primary HIV-1 infection. Cross-reactivity is attributable to the recognized epitopes being either invariant across clades or differing by single amino acid substitutions outside the HLA anchor sites. Semi-conservative and non-conservative substitutions that presumably involve the T-cell receptor contact sites have significant effects on T-cell recognition. Finally, further studies are needed to determine if the detection of cross-clade IFN-gamma T-cell responses indeed translates to cross-reactive antiviral activity. PMID- 17112645 TI - Small cell neuroendocrine carcinoma of the sinonasal region. A propose of a case. AB - We describe a 70-year-old man with rheumatoid arthritis and pulmonary fibrosis who presented with a month's history of pain in the left lateronasal region and inferior eyelid. On examination there was left exophthalmos, difficulty in coordinating eye movements, inflammation, erythema, and pain. Computed tomography showed a 3 cm mass in the left posterior ethmoid region, a biopsy specimen from which showed a small cell neuroendocrine tumour. He refused operation and was treated unsuccessfully with four cycles of cisplatin and etoposide. PMID- 17112646 TI - Treatment of specific phobia in adults. AB - This is a comprehensive review of treatment studies in specific phobia. Acute and long-term efficacy studies of in vivo exposure, virtual reality, cognitive therapy and other treatments from 1960 to 2005 were retrieved from computer search engines. Although specific phobia is a chronic illness and animal extinction studies suggest that relapse is a common phenomenon, little is known about long-term outcome. Treatment gains are generally maintained for one year, but longer follow-up studies are needed to better understand and prevent relapse. Acutely, the treatments are not equally effective among the phobia subtypes. Most phobias respond robustly to in vivo exposure, but it is associated with high dropout rates and low treatment acceptance. Response to systematic desensitization is more moderate. A few studies suggest that virtual reality may be effective in flying and height phobia, but this needs to be substantiated by more controlled trials. Cognitive therapy is most helpful in claustrophobia, and blood-injury phobia is uniquely responsive to applied tension. The limited data on medication have not been promising with the exception of adjunctive D clycoserine. Despite the acute benefits of in vivo exposure, greater attention should be paid to improve treatment acceptance and retention, and additional controlled studies of more acceptable treatments are needed. PMID- 17112647 TI - Examining the association between parenting and childhood anxiety: a meta analysis. AB - Theoretical models emphasize the role of parenting in the development and maintenance of child anxiety, but reviews of the empirical literature have provided mixed support for existing theories. To help clarify the role parenting plays in childhood anxiety, we conducted a meta-analysis of 47 studies testing the association between parenting and child anxiety. Across these studies, parenting accounted for only 4% of the variance in child anxiety. Moderator tests indicated that methodological factors (i.e., how child anxiety and parenting were conceptualized and assessed) may be a source of inconsistent findings within the literature. In addition, our analyses revealed that parental control was more strongly associated with child anxiety than was parental rejection. Specific subdimensions within parental rejection and control differed in their association with child anxiety (e.g., autonomy-granting accounted for 18% of the variance, but warmth <1%), indicating that efforts to disaggregate parenting dimensions may inform theory development and future research. Overall, however, the modest association between parenting and child anxiety suggests that understanding the origins of children's anxiety will require identifying factors other than parenting that account for the bulk of the variance. PMID- 17112648 TI - Small volume resuscitation with tempol is detrimental during uncontrolled hemorrhagic shock in rats. AB - BACKGROUND: In a previous study, titration of a hypertonic saline (HTS) solution during severe uncontrolled hemorrhagic shock (UHS) failed to reduce mortality. In a separate study, a novel antioxidant, polynitroxylated albumin (PNA) plus tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl), infused during shock increased long-term survival. We hypothesized that combining potent antioxidants with a hypertonic solution during UHS would preserve the logistical advantage of small volume resuscitation and improve survival. METHODS: An UHS outcome model in rats was used. UHS phase I (90 min) included blood withdrawal of 30 ml/kg over 15 min, followed by tail amputation for uncontrolled bleeding. At 20 min, rats were randomized to four groups (n=10 each) for hypotensive resuscitation from 20 to 90 min (mean arterial pressure [MAP] > or = 40 mmHg): HTS/starch group received 7.2% NaCl/10% hydroxyethyl starch; HTS/albumin group received 7.5% NaCl/20% albumin; HTS/PNA group received 7.5% NaCl/20% PNA; HTS/albumin+tempol group received 7.5% NaCl/20% albumin plus tempol. Resuscitation phase II (180 min) included hemostasis, return of shed blood and administration of fluids to restore MAP > or = 80 mmHg. Observation phase III was to 72 h. RESULTS: The total amount of fluid required to maintain hypotensive MAP during HS was low and did not differ between groups (range: 3.4+/-1.9 to 5.3+/-2.5 ml/kg). The rate of fluid administration required was higher in the HTS/albumin+tempol group compared to all other groups (p=0.006). Additional uncontrolled blood loss was highest in the HTS/PNA group (16.2+/-5.7 ml/kg [p=0.01] versus 10.4+/-7.9 ml/kg in the HTS/starch group, 7.7+/ 5.2 ml/kg in the HTS/albumin group and 8.2+/-7.1 ml/kg in the HTS/albumin+tempol group). MAP after start of resuscitation in phase I was lower in the HTS/albumin+tempol group than the HTS/albumin or HTS/PNA groups (p<0.01). This group was also less tachycardic. Long-term survival was low in all groups (2 of 10 after HTS/starch and 1 of 10 after HTS/albumin, 3 of 10 after HTS/PNA, 1 of 10 after HTS/albumin+tempol). Median survival time was shortest in the HTS/albumin+tempol group (72 min [CI 34-190]) compared to all other groups (p=0.01). CONCLUSIONS: Despite its benefits in other model systems, free tempol is potentially hazardous when combined with hypertonic fluids. PNA abrogates these deleterious effects on acute mortality but may lead to increased blood loss in the setting of UHS. PMID- 17112649 TI - Emergency management of arrhythmias and/or shocks in patients with implantable cardioverter defibrillators (ICDs). A statement on behalf of the Resuscitation Council (UK), Heart Rhythm UK (formerly The British Pacing and Electrophysiology Group, BPEG), The Joint Royal Colleges Ambulance Liaison Committee (JRCALC) and the Ambulance Services Association (ASA). PMID- 17112650 TI - Effect of instrument preference for operative deliveries on obstetrical and neonatal outcomes. AB - OBJECTIVES: To examine the relationship between physicians' instrument preference and obstetrical and neonatal outcomes. STUDY DESIGN: A retrospective cohort study comparing obstetrical and neonatal outcomes of second stage deliveries between obstetricians who prefer forceps (forceps >/=90%) with obstetricians with no preference to forceps (either instrument <90%) was completed using the McGill Obstetrical and Neonatal Database. Logistic regression analysis was used to obtain an adjusted odds ratio controlling for maternal, intrapartum and neonatal confounders. RESULTS: Two thousand and three hundred thirteen infants were delivered by 5 obstetricians who preferred forceps, and 9261 infants were delivered by 15 obstetricians with no instrument preference. Baseline characteristics were similar between the two groups. As compared to obstetricians who preferred forceps, obstetricians with no instrument preference had a higher rate of operative vaginal deliveries 1.5 (1.1-2.0), a higher cesarean section rate 2.5 (1.3-4.9) and a higher episiotomy rate in non-operative vaginal deliveries 3.4 (2.7-4.3). Infants delivered by obstetricians with no instrument preference were less likely to have significant bruising 0.3 (0.2-0.6) but more likely to have a cephalohematoma 3.0 (1.1-8.3). CONCLUSION: Physician instrument preference is an important determinant of outcomes that should be considered in studies evaluating instrumental deliveries. PMID- 17112651 TI - Genetic polymorphisms in vasoactive genes and preeclampsia: a meta-analysis. AB - There are controversies in reports on the association of polymorphisms in endothelial nitric oxide synthase, angiotensinogen, angiotensin receptor type 1 and angiotensin-converting enzyme genes with an increased risk of developing preeclampsia. We performed a systematic search of published case-control studies through the PubMed database up to January 2006, and report the results of a meta analysis of polymorphisms investigated in more than five studies: Glu298Asp in eNOS gene (9 analyses involving 1055 patients and 1788 controls), Met235Thr in AGT gene (13 analyses involving 1128 patients and 2278 controls), and intron 16 insertion-deletion polymorphism in ACE gene (10 analyses involving 1121 patients and 1361 controls). Statistically significant associations with preeclampsia were identified for the Met235Thr/AGT polymorphism: OR 1.65 (95% CI 1.19, 2.29) if the polymorphism is considered under the dominant genetic model, and OR 1.54 (95% CI 1.12, 2.11) under the recessive model. For insertion-deletion/ACE polymorphism, statistical significance was demonstrated when the polymorphism was considered under the recessive model: OR 1.51 (95% CI 1.17, 1.94). No single polymorphism was identified as having a major effect. PMID- 17112652 TI - Dynamical and structural properties of charged and uncharged lidocaine in a lipid bilayer. AB - Molecular dynamics computer simulations have been performed to investigate dynamical and structural properties of a lidocaine local anesthetic. Both charged and uncharged forms of the lidocaine molecule were investigated. Properties such as membrane area per lipid, diffusion, mass density, bilayer penetration and order parameters have been examined. An analysis of the lidocaine interaction with the lipid surrounding according to a simple mean field theory has also been performed. Almost all examined properties were found to depend on which of the two forms of lidocaine, charged or uncharged, is studied. The overall picture is a rather static behavior determined by the lipids for the charged molecules and more mobile situation of the uncharged form with higher diffusion and lower orientational and positional order. PMID- 17112653 TI - Volume and hydration changes of DNA-ligand interactions. AB - We report the volumetric and other thermodynamic properties of ethidium bromide (EB), propidium iodide (PI) and daunomycin (DAU) intercalating with poly(dA).poly(dT), poly[d(A-T)].poly[d(A-T)], and poly[d(G-C)].poly[d(G-C)], respectively, as well as minor groove binder Hoechst 33258 binding with poly[d(A T)].poly[d(A-T)]. The data were obtained using fluorescence titration and hydrostatic pressure measurements. Our thermodynamic data are combined with enthalpies from literature reports to analyze the thermodynamic characteristics of the different interactions. The differences are interpreted based on three processes related to hydration: I. burial of non-polar hydrophobic solvent accessible surface, II. burial of polar surface and formation of solute-solute H bonds, and III. disruption of "structural" hydration. Sequence dependent conformational changes may also be important when comparing ligand binding to different DNA sequences. We conclude that a combination of different thermodynamic parameters, especially volume change, is essential in order to understand the role of hydration in the energetics of DNA-ligand interactions. PMID- 17112654 TI - Kinetic mechanism of the Zn-dependent aryl-phosphatase activity of myo-inositol-1 phosphatase. AB - Myo-inositol-1-phosphatase (EC 3.1.3.25) is able to hydrolyze myo-inositol-1 phosphate in the presence of Mg(2+) ions at neutral pH, and also p-nitrophenyl phosphate in the presence of Zn(2+)-ions at acidic pH. This enzyme plays a role in phosphatidylinositol cell signalling and is a putative target of lithium therapy in manic depression. We elucidate here the kinetic mechanism of the Zn dependent activity of myo-inositol-1-phosphatase. As part of this analysis it was necessary to determine the basicity constants of p-nitrophenyl phosphate and the stability constant of its metal-complex in the presence of zinc chloride. We find that the Zn-dependent reaction may be described either by a rapid-equilibrium random mechanism or an ordered steady-state mechanism in which the substrate binds to the free enzyme prior to the metal ion. In both models the Zn-substrate complex acts as a high affinity inhibitor, yielding a dead-end species through its binding to the enzyme-Zn-substrate in rapid-equilibrium or to the enzyme phosphate complexes in a steady-state model. Phosphate is a competitive inhibitor of the enzyme with respect to the substrate and an uncompetitive inhibitor with respect to zinc ions. PMID- 17112655 TI - Effect of air bubbles in the coupling medium on efficacy of extracorporeal shock wave lithotripsy. AB - OBJECTIVES: Replacement of the water bath by a water cushion in newer lithotriptors introduces an acoustic interface and an ideal coupling agent is required to prevent energy loss at this interface. We aim to study the effect of bubbles in the coupling media on efficacy of extracorporeal shock wave lithotripsy (ESWL) by an in vitro experiment. METHODS: Using a standardised in vitro model 40 artificial stones were randomly treated on Modulith SLK lithotriptor using either conventional ultrasound gel (high bubble content) before and after displacing visible bubbles, a thin ultrasound gel (Therasonic) or silicon oil (both with negligible bubbles). Percentage area covered by bubbles in each case and the diameters and depth of crater created in each stone were measured by two blinded observers to determine the correlation between the bubble contents and disintegration efficacy. In vivo effect of two ultrasound gels was compared in terms of pain scores and stone fragmentation rates in ten patients treated with both gels. RESULTS: Volume of the craters was significantly greater with the Therasonic gel (102.4+/-33.4 mm3) or silicon oil (98.8+/-9.8 mm3) than the conventional ultrasound gel (49.2+/-32.6 mm3). But it was greatest (p<0.001) with ultrasound gel without bubbles (163.5+/-22.6 mm3). Depth and volume of the stone crater increased significantly with decreasing bubble contents of gel (p<0.001). Compared to standard ultrasound gel, patients treated with Therasonic gel reported significantly higher pain scores (median 3.5 vs. 8.0; p<0.001). CONCLUSIONS: Efficacy of ESWL is significantly correlated to air bubbles within the coupling gel and can be improved significantly by eliminating the bubbles from the coupling medium. PMID- 17112656 TI - Glucose-induced swelling in rat pancreatic alpha-cells. AB - Pancreatic beta-cells increase in volume when exposed to elevated concentrations of extracellular glucose. This study has examined the effects of glucose on the volumes of pancreatic alpha-cells, which like beta-cells are regulated by glucose, and intestinal epithelial Caco-2 cells which are unresponsive to glucose. Cell volume changes were monitored by a video-imaging method. Increasing the extracellular glucose concentration caused a concentration-dependent increase in alpha-cell volume over the range 1-20mM. Glucose-induced swelling was not, however, observed in Caco-2 cells. The glucose-induced swelling in both alpha- and beta-cells was abolished by 0.5mM phloretin, an inhibitor of the GLUT proteins, indicating that GLUT mediated glucose transport is a pre-requisite for swelling. Glucose metabolism also appears to be essential, as islet cell swelling was not observed with 16 mM 3-O-methyl glucose. These data suggest that glucose induced swelling may be a property exclusive to glucose-regulated cells. PMID- 17112657 TI - The influence of ice formation on vaporization of LNG on water surfaces. AB - The spillage of LNG on water surfaces can lead, under certain circumstances, to a decrease in the surface temperature of water and subsequent freezing. A model for heat transfer from water to LNG is proposed and used to calculate the surface temperature of water and examine its influence on the vaporization rate of LNG. For this purpose LNG was modeled based on the properties of pure methane. It was concluded that when LNG spills on a confined, shallow-water surface the surface temperature of water will decrease rapidly leading to ice formation. The formation of an ice layer, that will continue to grow for the duration of the spill, will have a profound effect upon the vaporization rate. The decreasing surface temperature of ice will decrease the temperature differential between LNG and ice that drives the heat transfer and will lead to a change of the boiling regime. The overall effect would be that the vaporization flux would first decrease during the film boiling; followed by an increase during the transition boiling and a steady decrease during the nucleate boiling. PMID- 17112658 TI - Modeling the release, spreading, and burning of LNG, LPG, and gasoline on water. AB - Current interest in the shipment of liquefied natural gas (LNG) has renewed the debate about the safety of shipping large volumes of flammable fuels. The size of a spreading pool following a release of LNG from an LNG tank ship has been the subject of numerous papers and studies dating back to the mid-1970s. Several papers have presented idealized views of how the LNG would be released and spread across a quiescent water surface. There is a considerable amount of publicly available material describing these idealized releases, but little discussion of how other flammable fuels would behave if released from similar sized ships. The purpose of this paper is to determine whether the models currently available from the United States Federal Energy Regulatory Commission (FERC) can be used to simulate the release, spreading, vaporization, and pool fire impacts for materials other than LNG, and if so, identify which material-specific parameters are required. The review of the basic equations and principles in FERC's LNG release, spreading, and burning models did not reveal a critical fault that would prevent their use in evaluating the consequences of other flammable fluid releases. With the correct physical data, the models can be used with the same level of confidence for materials such as LPG and gasoline as they are for LNG. PMID- 17112659 TI - Integration of electrokinetics and chemical oxidation for the remediation of creosote-contaminated clay. AB - Remediation of clayey soils that are contaminated with polycyclic aromatic hydrocarbons (PAHs) is a challenging task that may require integration of several technologies. The benefits of integrating in situ electrokinetic remediation with chemical oxidation were evaluated in laboratory-scale experiments lasting for 8 weeks. A voltage gradient of 48 V/m of direct current and 4.7 V/m of alternating current and periodic additions of chemical oxidants were applied to creosote contaminated soil. Electrokinetically enhanced oxidation with sodium persulphate resulted in better PAH removal (35%) than either electrokinetics (24%) or persulphate oxidation (12%) alone. However, the improvement was shown only within 1/3 (5 cm) of the soil compartment. Electrokinetics did not improve the performance of Fenton oxidation. Both chemical oxidants created more positive oxidation-reduction potential than electrokinetic treatment alone. On the other hand, persulphate treatment impaired the electroosmotic flow rate. Elemental analyses showed reduction in the natural Al and Ca concentrations, increase in Zn, Cu, P and S concentrations and transfer of several metal cations towards the cathode. In conclusion, the results encourage to further optimisation of an integrated remediation technology that combines the beneficial effects of electrokinetics, persulphate oxidation and Fenton oxidation. PMID- 17112660 TI - Removal of phenol and 4-chlorophenol by surfactant-modified natural zeolite. AB - In this study the adsorption characteristics of phenol and 4-chlorophenol by surfactant-modified zeolite was investigated. Batch studies were performed to evaluate the effects of various experimental parameters such as contact time, adsorbent dose, initial concentration, and temperature on the removal of phenol and 4-chlorophenol. The sorption kinetics was tested for intraparticle diffusion, Elovich, and pseudo-second order reaction and rate constants of kinetic models were calculated. Equilibrium isotherms for the adsorption of phenol were analyzed by Freundlich, Langmuir, and Tempkin isotherm models. Freundlich isotherm was found to best represent the data for phenol and 4-chlorophenol adsorption. PMID- 17112661 TI - Extraction of heavy metals from municipal solid waste incinerator (MSWI) bottom ash with organic solutions. AB - Municipal solid waste incinerator (MSWI) bottom ash often cannot be recycled as construction material in Flanders, because leaching of Cu exceeds the limit value of 0.5mg/kg. Leaching of other components such as Mo and Sb is critical as well, but limit values for these elements are to date only informal. A treatment technique was investigated to lower pollutant leaching: extraction with solutions of organic complexants to remove Cu. Six different solutions were used, of which washing with citric acid and ammonium citrate decreases Cu leaching to below the limit value. Extraction was then performed with different concentrations of ammonium citrate. Subsequent washing of the extracted material with distilled water appears to be vital to remove all residual ammonium citrate. Extraction with a 0.2M solution of ammonium citrate followed by three washing steps decreases metal leaching to below the limit values. PMID- 17112662 TI - Optimized photocatalytic degradation of Alcian Blue 8 GX in the presence of TiO2 suspensions. AB - The photocatalytic degradation of Alcian Blue 8 GX, a cationic copper phthalocyanine dye, has been investigated in aqueous suspensions containing the commercial catalyst TiO(2) P-25. The photodegradation of the organic molecule follows approximately a pseudo-first kinetic order, according to the Langmuir Hinshelwood model. The effect of catalyst concentration, pH of the initial solution and the H(2)O(2) concentration upon the reaction rate was ascertained. It was shown that the photocatalytic degradation reaction can be mathematically described as a function of parameters such as pH, H(2)O(2) concentration and irradiation time, being modeled by the use of the response surface methodology. Optimized values for oxidizing agent, concentration, pH and UV exposure time for the studied system were determined. PMID- 17112663 TI - Impact of substrates and cell immobilization on siderophore activity by Pseudomonads in a Fe and/or Cr, Hg, Pb containing-medium. AB - To increase the amount of bioavailable metals in phytoextraction purposes, soil bioaugmentation with Pseudomonads, as siderophore producers with high metal complexation levels, could be relevant. Unfortunately, siderophore synthesis may be inhibited by soluble iron in soil and bacteria can suffer at the same time from the toxicity of some other metals, predation and oligotrophy. To overcome these drawbacks, we attempted to co-locate a carbon substrate and Pseudomonas aeruginosa or P. fluorescens in Ca-alginate beads. First, free-cell cultures showed that glycerol, fructose, mannitol and skim milk enhanced the siderophore activity which was the highest in the medium with neither Fe or TM (toxic metal) (Cr, Hg and Pb) and the lowest in the Fe-containing medium without TM. The negative effect of iron was partly offset when TM was added to the medium. In a second part, co-location of microorganisms and substrates was only feasible with skim milk. By comparison with free cells, siderophore activity by immobilized cells was higher in culture media containing Fe with or without TM (up to a ratio of 9), and varied in a narrow margin, according to the medium composition. PMID- 17112664 TI - Vagal afferents mediate the feeding response to mercaptoacetate but not to the beta (3) adrenergic receptor agonist CL 316,243. AB - To evaluate the role of subdiaphragmatic vagal afferents in the anorectic response to peripheral administration of the highly selective beta(3)-adrenergic receptor agonist CL 316,243 (CL), we tested the ability of intraperitoneal (IP) injections of CL to inhibit feeding in rats with subdiaphragmatic vagal deafferentation (SDA, n=13) or sham surgeries (SHAM, n=13). Doses of 10, 100 and 1000 ng/kg CL significantly reduced feeding by statistically similar amounts in SHAM and SDA rats. One hour after IP injection, each dose of CL also significantly increased plasma concentrations of free fatty acids and beta hydroxybutyrate, an indicator of hepatic fatty acid oxidation (FAO), whereas 6h after injection only the two highest CL doses increased plasma beta hydroxybutyrate. In contrast, peripheral administration of the FAO inhibitor mercaptoacetate (MA, 45.6 mg/kg IP) stimulated feeding in SHAM but not in SDA rats, extending previous data suggesting a necessary role of vagal afferents in the feeding-stimulatory effect of FAO inhibition. We conclude that subdiaphragmatic vagal afferents are essential for the feeding-stimulatory action of MA but not for the anorectic action of peripheral CL and that CL-induced increase in hepatic FAO is not essential for its feeding-inhibitory effect. PMID- 17112665 TI - Rate of neurite outgrowth in sympathetic neurons is highly resistant to suppression of protein synthesis: role of protein degradation/synthesis coupling. AB - Neurites projecting to their target tissues during embryogenesis are subject to many perturbations that could influence their rate of growth. For example, environmental influences such as supply of neurotrophic factor or electrical activity profoundly influence the rate of neuronal protein synthesis. Because accumulation of protein is necessary for outgrowth to proceed normally, a perturbation in protein synthesis could cause a net change in the rate of accumulation of proteins with the result that neurite outgrowth rate increases or decreases. That neurite outgrowth does not normally seem to be subject to such perturbations suggests involvement of a homeostatic system controlling the rate of outgrowth. Consistent with this hypothesis, we show here that the rate of growth of neurites of sympathetic neurons is highly resistant to decreased rates of protein synthesis. Chronic suppression of protein synthesis by 60% had no significant effect on neurite outgrowth over a 2-day period while complete suppression halted it almost immediately. By the 3rd day of exposure, 60% suppression slowed outgrowth. Sustained suppression of protein synthesis rate by 33% had no effect on rate of outgrowth even after 7 days. We show that the ability of the growing neurites to resist protein synthesis suppression appears to be caused, at least in part, by a parallel decrease in the rate of protein degradation. The result of this coupling between degradation and synthesis is that proteins can continue to accumulate even when protein synthesis rate decreases, allowing normal rates of neurite outgrowth. PMID- 17112666 TI - Myocyte enhancing factor-2A in Alzheimer's disease: genetic analysis and association with MEF2A-polymorphisms. AB - Polymorphisms at different genes have been proposed as determinants of the risk for developing late-onset Alzheimer's disease (LOAD). Among the several candidate genes are those that encode proteins involved in neuronal degeneration/survival. Studies of primary neuronal cultures supported that members of the myocyte enhancing factor-2 (MEF2) family of transcription factors have an anti-apoptotic effect, regulating the expression of proteins involved in neuronal survival and differentiation. We analysed the MEF2A gene in a total of 357 patients (mean age 72 years, range 60-97 years). Among others, a Pro279Leu in exon 8 and a polyglutamine (CAG) repeat polymorphisms in exon 12 were found. These variants were also genotyped in 495 healthy controls (>50 years old), and the frequencies were statistically compared. Eight patients were 279L (six P/L and two L/L), compared to only one control (2% vs. 0.2%; p=0.004, OR=11.32). There was a significantly higher frequency of 279L-carriers among APOE epsilon4+ (7/154=4.5%), compared to epsilon4- (1/203) (p=0.02). In conclusion, our work suggests that the variation at the MEF2A gene could be involved in the risk of developing LOAD. Because MEF2 has been related with neuronal survival, and the 279L allele has been related with a reduction in the transcriptional activation activity of MEF2A, the effect of this allele could be mediated through a down regulation of antiapoptotic genes. PMID- 17112667 TI - Cases of reduced cyathostomin egg-reappearance period and failure of Parascaris equorum egg count reduction following ivermectin treatment as well as survey on pyrantel efficacy on German horse farms. AB - In 2003 and 2004, on a total of 63 different German horse farms, a survey using the faecal egg count reduction (FECR) test was performed to investigate the efficacy of ivermectin (IVM, Ivomec) and pyrantel (PYR, Banminth) treatment against gastro-intestinal nematodes in a total of 767 horses. IVM treatment resulted in 100% reduction of the cyathostomin egg production 14 and 21 days post treatment (d.p.t.) on 37 farms. On the remaining five farms, the mean faecal egg count reduction ranged between 97.7 and 99.9%. The mean cyathostomin FECR following PYR treatment ranged between 92.2 and 100% on the 25 farms tested. Therefore, based on the 90% FECR threshold suggested for detection of anthelmintic resistance in horses, neither IVM nor PYR anthelmintic resistance was detected. However, if the thresholds recommended for the detection of resistance in small ruminants were applied, on one and four farms signs of reduced IVM and PYR efficacy, respectively, were observed. In 2005, to further investigate these findings, the cyathostomin egg-reappearance period (ERP) following IVM treatment was examined on six selected farms, two of which were found to show less than 99.8% FECR in the previous survey. On these two latter farms, the ERP was less than 5 weeks, while on the other four it was at least 8 weeks. Earlier investigations described IVM cyathostomin ERP of at least 9 weeks. The efficacy of IVM to reduce Parascaris equorum egg excretion was also studied. On one farm in 2 consecutive years, IVM treatment did not lead to a significant reduction in P. equorum faecal egg counts in one and five young horses, respectively. PMID- 17112668 TI - Prevalence of Giardia duodenalis and Cryptosporidium spp. in dogs from different living conditions in Uberlandia, Brazil. AB - Infection rates with Giardia duodenalis and Cryptosporidium spp. were compared among dogs living under different conditions. Stool samples (n = 433) collected from dogs of different ages, gender, living conditions and origin were analyzed using three techniques, i.e., centrifugal flotation in zinc sulfate solution, centrifugal flotation in sucrose solution, and methylene blue gram safranin staining. Eighty-nine of the samples were from stay dogs living in shelters run by animal protection societies, 199 were from kennels and 122 from households. A total of 119 (29.0%) had G. duodenalis cysts and six (1.4%) were positive for Cryptosporidium spp. oocysts. Dogs from kennels were most frequently affected by G. duodenalis (49.7%) while those from shelters showed a higher prevalence of Cryptosporidium spp. (2.2%). A significant difference (p < or = 0.05) was observed between immature dogs and adults only with respect to Giardia infection. There was no significant difference between the gender with regard to the presence of either protozoan. PMID- 17112669 TI - The influence of distance on movement of tabanids (Diptera: Tabanidae) between horses. AB - Two studies evaluated the potential use of spatial barriers to reduce the mechanical transmission of disease agents by tabanids in the Pantanal region of Brazil. Tabanids at stations separated by four different distances (5, 10, 25, and 50m) were marked. In the first study, tabanids were marked and allowed to feed until engorgement or natural interruption occurred and captured if they transferred to the other horse. A total of 2847 tabanids belonging to nine different species were marked. The percentage of tabanids that moved between horses was 10.5 at 5m, 6.8 at 10m, and 4.6 at 25m. In the second study, flies were marked, feeding was then interrupted, and the flies were released approximately 50cm from the host. A total of 1274 tabanids belonging to five different species were marked. The percentage of flies that moved between horses was 9.7 at 5m, 9.7 at 10m, and 4.6 at 25m. No tabanids transferred between animals separated by 50m in either experiment. The results of this study strongly support the recommendation that segregation of animals effectively prevents the mechanical transmission of pathogens by tabanids. PMID- 17112670 TI - Presence of Neospora caninum specific antibodies in three dairy farms in Georgia and two in Texas. AB - Neospora caninum is known to cause abortion in cattle. This study demonstrated the presence of specific IgG to Neospora in milk and serum samples obtained from three dairy farms in Georgia and two in Texas. Samples from four hundred fourteen dairy cows were examined using a western blot assay of which 362 were milk and 87 were serum. Samples with antibodies to Neospora were identified in 32.1% (105/327) of the examined animals in Georgia, whereas in Texas it was identified in 10.3% (9/87). Positive Georgia samples were found in 24.4% from farm A (28/115), 21.6% from farm B (30/139), and 64.4% from farm C (47/73). In Texas, 13.5% (7/52) of animals in farm D and 5.71% (2/35) from farm E also had specific antibodies to Neospora. The number of animals from Georgia dairy farms with antibodies to Neospora was significantly higher than the Texas dairy farms. This may be related to the age of the animals examined in this study (more than 2 years old). Antibodies present in sera had excellent agreement with the antibodies present in milk. Collection of milk samples for serological testing is easier and less invasive than obtaining bovine sera, therefore offering an alternative for animal testing. PMID- 17112671 TI - Infectivity rate and transmission potential of Hyalomma anatolicum anatolicum ticks for Babesia equi infection. AB - The infectivity rate of Babesia equi in the salivary glands of Hyalomma anatolicum anatolicum was assessed. The hungry nymphs were fed on a donkey experimentally infected with B. equi. The engorged dropped-off nymphs were collected at different levels of parasitaemia and kept in BOD incubator. After ecdysis, the hungry adults were prefed on rabbits for different time intervals, thereafter the salivary glands were dissected out and acini were examined after methyl green pyronin (MGP) staining. A total of 134 male and 139 female ticks were dissected out. Average infected acini per tick were found to be significantly higher (p<0.05) in male as compared to the female ticks. Further, maximum infected acini in both male and female ticks were found at 24h of prefeeding on rabbits and overall infected acini per tick increased with rise in parasitaemia. The release of infected ticks on susceptible donkeys resulted in development of clinical babesiosis. PMID- 17112672 TI - TRB3 interacts with CtIP and is overexpressed in certain cancers. AB - TRB3, a human homolog of Drosophila Tribbles, has been recently shown as a critical negative regulator of Akt and S6 kinase activation in a number of cellular processes. Here we found that TRB3 interacted with an important cell cycle regulator CtIP (CtBP-interacting protein) and the interaction involved the C-terminus of both proteins. Interestingly, TRB3 and CtIP co-localized to the nucleus in HeLa cells and exhibited a unique dot-like pattern. Finally, we demonstrated that TRB3 was overexpressed in multiple tumor tissues. Since CtIP plays important roles in cell cycle checkpoint control and it has been implicated in tumorigenesis, our data suggest that TRB3 may be involved in these biological processes through interacting with CtIP. PMID- 17112673 TI - Effects of different chemotherapy regimens on survival for advanced cervical cancer: systematic review and meta-analysis. AB - BACKGROUND: A large number of trials have assessed various chemotherapy regimens for the treatment of advanced cervical cancer, but there is uncertainty about the magnitude of survival benefits. METHODS: We searched (last update January 2006) for trials in women with locally advanced or disseminated cervical cancer that compared neo-adjuvant or concurrent chemotherapy plus radiotherapy versus radiotherapy alone; or different chemotherapy regimens among themselves (with or without background radiotherapy in both arms). Sixty-five trials were identified with survival data on 11,180 women. Results for survival were combined with fixed and random effects models and between-study heterogeneity was estimated. Separate results were obtained for different regimens, cycle length, and type of chemotherapy (neo-adjuvant, concurrent, without radiotherapy). RESULTS: Twenty two comparisons had survival data on 3837 women randomized to receive chemotherapy plus radiotherapy versus radiotherapy alone; the summary relative hazard for mortality was 0.95, 95% CI, 0.83-1.08. Modest between-study heterogeneity (I2=38%) seemed to be due to contradictory results in early trials; trials published in the last decade had a summary relative hazard 0.89 (95% CI, 0.78-1.02) and no between-study heterogeneity (I2=0%). Results were similar for neo-adjuvant chemotherapy and for concurrent chemo-radiotherapy. Cisplatin or cisplatin-based combinations had no significant benefit overall, but a potential benefit was seen with short-length cycles (14 days) and a marginally significant harm with longer-length cycles (summary relative hazards 0.80, 95% CI, 0.66-0.99 and 1.18, 95% CI, 1.02-1.38, respectively). The summary relative hazard was 1.02, (95% CI, 0.84-1.24) for trials using neo-adjuvant chemotherapy and 0.85 (95% CI, 0.73-1.00) for trials using concurrent chemotherapy. CONCLUSIONS: Evidence on chemotherapy in women with advanced cervical cancer is not encouraging for major survival benefits. However, small benefits have been observed in some trials, especially with short-length cycles of cisplatin-based regimens and concurrent chemotherapy and radiotherapy. PMID- 17112674 TI - Darbepoetin alfa (Aranesp) improves recognition memory in adult rats that have sustained bilateral ventral hippocampal lesions as neonates or young adults. AB - Recognition memory was assessed in adult rats that received bilateral injections of saline (sham lesions) or ibotenic acid (lesioned) in the ventral hippocampus as neonates (postnatal day 7, PD7) or young adult (42 days of age, PD42) using the Novel Object Recognition Test (NORT). Normal or sham-lesioned rats were able to distinguish novel from familiar objects over a 0.5 and 2 h delay between the sample and choice phases. Adult rats (PD70) lesioned as neonates performed progressively worse than sham-lesioned animals at delays of 0.5 and 2 h. A single injection of darbepoetin alfa (500 or 5000 U/kg, i.p.), given 1 h before the sample phase restored performance 0.5 or 2 h later in the choice phase to same levels as sham-lesioned rats. Adults lesioned on PD42 displayed deficits in NORT performance with a 2 h delay between the choice and sample phases that were completely reversed by administration of darbepoetin alfa (5000 U/kg, i.p.) 1 h before the sample phase. These results suggest that darbepoetin alfa may have utility in treating memory deficits associated with brain dysfunction related to developmental disorders such as schizophrenia. PMID- 17112675 TI - Phase relationships support a role for coordinated activity in the indirect pathway in organizing slow oscillations in basal ganglia output after loss of dopamine. AB - The goal of the present study was to determine the phase relationships of the slow oscillatory activity that emerges in basal ganglia nuclei in anesthetized rats after dopamine cell lesion in order to gain insight into the passage of this oscillatory activity through the basal ganglia network. Spike train recordings from striatum, subthalamic nucleus (STN), globus pallidus (GP), and substantia nigra pars reticulata (SNpr) were paired with simultaneous local field potential (LFP) recordings from SNpr or motor cortex ipsilateral to a unilateral lesion of substantia nigra dopamine neurons in urethane-anesthetized rats. Dopamine cell lesion induced a striking increase in incidence of slow oscillations (0.3-2.5 Hz) in firing rate in all nuclei. Phase relationships assessed through paired recordings using SNpr LFP as a temporal reference showed that slow oscillatory activity in GP spike trains is predominantly antiphase with oscillations in striatum, and slow oscillatory activity in STN spike trains is in-phase with oscillatory activity in cortex but predominantly antiphase with GP oscillatory activity. Taken together, these results imply that after dopamine cell lesion in urethane-anesthetized rats, increased oscillatory activity in GP spike trains is shaped more by increased phasic inhibitory input from the striatum than by phasic excitatory input from STN. In addition, results show that oscillatory activity in SNpr spike trains is typically antiphase with GP oscillatory activity and in phase with STN oscillatory activity. While these observations do not rule out additional mechanisms contributing to the emergence of slow oscillations in the basal ganglia after dopamine cell lesion in the anesthetized preparation, they are compatible with 1) increased oscillatory activity in the GP facilitated by an effect of dopamine loss on striatal 'filtering' of slow components of oscillatory cortical input, 2) increased oscillatory activity in STN spike trains supported by convergent antiphase inhibitory and excitatory oscillatory input from GP and cortex, respectively, and 3) increased oscillatory activity in SNpr spike trains organized by convergent antiphase inhibitory and excitatory oscillatory input from GP and STN, respectively. PMID- 17112676 TI - Selective disarrangement of the rostral telencephalic cholinergic system in heterozygous reeler mice. AB - Reelin (RELN) is a key molecule for the regulation of neuronal migration in the developing CNS. The reeler mice, which have spontaneous autosomal recessive mutation in the RELN gene, reveal multiple defects in brain development. Morphological, neurochemical and behavioral alterations have been detected in heterozygous reeler (HR) mice, suggesting that not only the presence, but also the level of RELN influences brain development. Several studies implicate an involvement of RELN in the pathophysiology of neuropsychiatric disorders in which an alteration of the cholinergic cortical pathways is implicated as well. Thus, we decided to investigate whether the basal forebrain (BF) cholinergic system is altered in HR mice by examining cholinergic markers at the level of both cell body and nerve terminals. In septal and rostral, but not caudal, basal forebrain region, HR mice exhibited a significant reduction in the number of choline acetyltransferase (ChAT) immunoreactive (ir) cell bodies compared with control mice. Instead, an increase in ChAT ir neurons was detected in lateral striatum. This suggests that an alteration in ChAT ir cell migration which leads to a redistribution of cholinergic neurons in subcortical forebrain regions occurs in HR mice. The reduction of ChAT ir neurons in the BF was paralleled by an alteration of cortical cholinergic nerve terminals. In particular, the HR mice presented a marked reduction of acetylcholinesterase (AChE) staining accompanied by a small reduction of cortical thickness in the rostral dorsomedial cortex, while the density of AChE staining was not altered in the lateral and ventral cortices. Present results show that the cholinergic basalo-cortical system is markedly, though selectively, impaired in HR mice. Rostral sub-regions of the BF and rostro-medial cortical areas show significant decreases of cholinergic neurons and innervation, respectively. PMID- 17112677 TI - Temporal profile of connexin 43 expression after photothrombotic lesion in rat brain. AB - Following focal ischemic injury, several mechanisms lead to secondary expansion of the affected area and therefore increase the initial damage. We thoroughly investigated the expression of astrocytic connexin 43 (Cx43) after photothrombosis in rat brain. The temporal profile of Cx43 mRNA as well as protein expression was studied in remote, structurally uninjured cortical and hippocampal areas. The hippocampal formation revealed an increased number of Cx43 mRNA positive astrocytes and an up-regulated protein expression exclusively in the ipsilateral stratum oriens. We assume a participation of this region in glia scar formation. While Cx43 mRNA positive cells were transiently increased, immunoreactivity was reduced in the somatosensory cortex of injured hemispheres. The observed decrease of Cx43 protein in the post-ischemic cerebral cortex implies an impairment of gap junctional intercellular communication which might be detrimental to the brain. PMID- 17112678 TI - Mitochondrial involvement in transhemispheric diaschisis following hypoxia ischemia: Clomethiazole-mediated amelioration. AB - Mitochondria play a central role in both the physiological and pathophysiological regulation of cell survival/death. Increasing evidence places mitochondrial dysfunction at the center of many neuropathological conditions. The present study investigates the extent of mitochondrial dysfunction in cortical, hippocampal and cerebellar tissues in a rat model of hypoxia-ischemia (HI). We hypothesized that; mitochondrial dysfunction in situ may be prevented by treatment with clomethiazole (CMZ), a GABA(A) receptor agonist. Assessment of mitochondrial FAD linked respiration at both 1- and 3-day post-HI revealed a marked decrease in activity from ipsilateral cortical and hippocampal regions (P<0.001). In addition, small changes were seen in contralateral cortical and hippocampal tissues as well as in the cerebellum at 3-days (P<0.05). Assessment of the mitochondrial electron transport chain (complexes I-V), and mitochondrial markers of integrity (citrate synthase) and oxidative stress (aconitase) confirmed mitochondrial impairment in ipsilateral regions following HI. Complexes I, II III, V and citrate synthase were also impaired in contralateral regions and cerebellum 3-days post-HI. Treatment with CMZ (414 mg/kg/day via minipumps) provided marked protection to all aspects of neuronal tissue assessed. Circulating cytokine (interleukin [IL]-1alpha, IL-1beta, tumor necrosis factor [TNF]-alpha, granulocyte macrophage colony-stimulating factor [GM-CSF], IL-4 and IL-10) levels were also assessed in these animals 3-days post-HI. Plasma IL 1alpha, IL-1beta, TNF-alpha and GM-CSF levels were significantly increased post HI. Treatment with CMZ ameliorated the increases in IL-1alpha, IL-1beta, TNF alpha and GM-CSF levels while increasing plasma IL-4 and IL-10 levels. This study provides evidence of the extent of mitochondrial damage following an HI-insult. In addition, we have shown that protection afforded by CMZ extends to preservation of mitochondrial function and integrity via anti-inflammatory mediated pathways. PMID- 17112679 TI - Potential therapeutical effects of cannabidiol in children with pharmacoresistant epilepsy. PMID- 17112680 TI - Human chorionic gonadotropin as an angiogenic factor in breast cancer during pregnancy. AB - Breast cancer associated with pregnancy is defined as the one in which the diagnosis is made in a pregnancy or within one year of delivery. Breast cancer is the second most common malignancy during pregnancy and it is generally considered to have a worse prognosis than the one that is not associated with pregnancy. The average patient is between 32 and 38 years of age. Steroid hormone receptor positive cell populations comprise 80% of breast cancers, however, estrogen receptor levels in pregnancy-associated tumors are often low or absent. Extensive laboratory data suggest that angiogenesis plays an essential role in breast cancer development, invasion, and metastasis. One of the most powerful stimulatory factors, vascular endothelial growth factor (VEGF), functions in autocrine/paracrine pathways. Current research, generally has validated the poor prognosis and early relapse that are associated with increasing microvessel density, which is related to VEGF expression in tumoral cells. During pregnancy, human chorionic gonadotropin (hCG) induces neovascularization in various tissues, one of them being the placenta. Its receptors have been detected in epithelial cells in breast carcinoma tissue, and breast cancer cell lines. According to this premise the hCG normally produced during pregnancy could induce the synthesis of VEGF and by this means stimulate the development and metastatic potential of breast cancer cells in the pregnancy period. Thus, research involving hCG and VEGF would help us understand the physiopathology of breast cancer during pregnancy, as well as provide us with probable prognostic tools. PMID- 17112681 TI - The role of methemoglobinemia in early and late complicated pregnancy. AB - The objective of this review was to direct attention about methemoglobin as a biomarker which has an important role in the detection of adverse effects of the oxidative stress, misbalanced production of ROS, RNS and RSS. According to our hypothesis, a pregnant woman continuously inhaling environmental toxics as fuel burning products, will traverse three, not two from current thought, distinct stages. The main difference among present three-stage hypothesis and other hypotheses is the assertion that, in the pathogenesis of early and late complicated pregnancy, methemoglobin takes on an important role. Secondly, we also observed the utero-placental changes as "locus manifesting minoris resistentiae" in complicated pregnancy are not the causes but a consequence of increased systemic oxidative stress. Methemoglobin and hemolysis both occur as a result of oxidative stress, but the prevalent difference between them is that methemoglobin is a reversible phenomenon (oxidant-antioxidant balance) whereas hemolysis, which occurs as a result of oxidative stress on the erythrocyte membrane, is an irreversible event. Methemoglobinemia can additionally exacerbate an existing anemia, stimulating hypoxia that may be dangerous for both mother and fetus. Own prospective study of methemoglobin in pregnancy, revealed a significant rise in the level of methemoglobin >1.5 g/L (r=0.72, p<0.01) in the exposure period, which can be explained on the basis of an oxidant-antioxidant imbalance, resulting in methemoglobinemia. Methemoglobinemia and stillbirth recorded throughout exposure period are significantly higher than those recorded in the control period (p=0.0205), and the frequencies of reproductive loss were significantly lower in the control than in the exposure period (p<0.05). Results suggest that methemoglobin as individual indicator of oxidative stress is an early marker of the identification of women with a pregnancy risk. It has the advantage of being applicable some time before ultrasonic examination becomes feasible. Further support for this assumption will require further investigations that may lead to the supposition that increasing level of methemoglobin is related to environmental toxicities complicated pregnancy and IUGR, preeclampsia, and a high percentage of perinatal mortality and morbidity. PMID- 17112682 TI - A hypothesis regarding complement activation and amniotic fluid embolism. AB - Amniotic fluid embolism, a rare, sudden and often fatal illness of pregnancy may not be a true embolic event resulting from the physical obstruction of the pulmonary vasculature. The high degree of variability in symptoms, the lack of characteristic findings on radiological exam, the absence of a dose-response effect on symptoms, and the occasional occurrence of coagulopathies are not entirely consistent with a physical block to the circulation as the main mechanism of disease. Alternatively, it might be the result of complement activation initiated by fetal antigen leaking into the maternal circulation. This rare immune response may be initiated by a rare pathological antigen, or by common antigens presented uncommonly--in amount, timing, or frequency of entry into the maternal circulation. Some very early evidence in AFE patients supports this hypothesis but is not conclusive. Complement levels remain well within the normal range during uncomplicated parturition. A prior theory that AFE might be a result of maternal anaphylaxis to fetal antigen has much less evidence to support it. The disseminated intravascular coagulation often seen in this and other serious obstetrical illnesses may be a secondary result of complement activation rather than the direct introduction of pro-coagulants into the maternal circulation although the link between the complement and coagulation pathways, if any, remains poorly defined. Through currently available laboratory testing, both the complement hypothesis and the anaphylaxis mechanism are able to be assessed. Direct measurement of serum complement as well as serum tryptase and urinary histamine are readily obtained tests in community hospitals as well as tertiary care hospitals. If the hypothesis proves true, this investigation may be of profound importance to understanding immune tolerance. Rather, than asking why one pregnant woman in 20,000 develops a violent immune reaction to the fetus, a better question is why do not all pregnant women reject the fetus which is a large collection of foreign antigens? PMID- 17112683 TI - Parikh's formula to minimize errors in calculating expected date of delivery. PMID- 17112684 TI - [Using a 48-hour delay from admission to the first positive culture without any other consideration: an accurate method to differentiate acquired and imported methicillin-resistant Staphylococcus aureus?]. AB - To differentiate imported and acquired strains of methicillin-resistant Staphylococcus aureus (MRSA), a 48-hour delay from hospital admission to the first MRSA-positive culture is usually considered. To assess if taking into account this delay without any other consideration is an accurate method, we defined 3 situations for whom we considered the MRSA acquisition status as questionable. The other situations were defined as either acquired MRSA or imported MRSA. We determined the acquisition status of MRSA (acquired, imported, or questionable) isolated during a 20-month period by considering or not considering screening samples performed on admission. The ratio "imported MRSA/acquired MRSA" (I/A) was calculated according to (1) the consideration of MRSA with questionable status as imported or acquired, and (2) the consideration of screening samples or not in the calculation of the ratio. The acquisition status in our hospital was questionable in 3.6% of patients when all samples were considered and in 12,0% when only clinical samples were taken into account (p = 0,01). The ratio I/A was 4-fold higher by considering both clinical and screening cultures and questionable status as imported than by considering only clinical samples and questionable status as acquired. Using a 48-hour delay without any other consideration is probably an accurate method to differentiate acquired and imported MRSA when a selective screening programme at admission in operational. Conversely, this definition seems to be more hazardous in the absence of screening. PMID- 17112685 TI - Molecular evolution of PIII-SVMP and RGD disintegrin genes from the genus Crotalus. AB - Several types of disintegrins have been isolated from Crotalus spp rattlesnakes, including RGD disintegrins, and PIII-SVMPs. We isolated six cDNAs from snake venom glands using RT-PCR. Three RGD disintegrins (atroxatin, mojastin, and viridistatin) and three PIII-SVMPs (catroriarin, scutiarin, and viristiarin) cDNAs were isolated from the rattlesnakes Crotalus atrox, Crotalus scutulatus scutulatus, and Crotalus viridis viridis, respectively. Atroxatin and Viridistatin shared 90% amino acid identity to each other, and 87% identity to Mojastin. Scutiarin and Viristiarin were identical. All PIII-SVMPs isolated in this study shared the highest amino acid identity with Catrocollastatin. cDNA and protein sequences for RGD disintegrins, one MVD disintegrin, and PIII-SVMPs of the genus Crotalus (present in the NCBI database), were used in phylogenetic analysis. Neighbor-joining analysis of PIII-SVMP and RGD/MVD disintegrin-coding DNA sequences showed that these groups of genes separate into separate clades. A Phi(ST) pairwise comparison and Analysis of Molecular Variance (AMOVA) between PIII-SVMPs and RGD/MVD disintegrins showed significant genetic differences. Mutations observed in ten of the cDNAs analyzed did not affect Cys-coding sequences. Our K(A)/K(S) data suggest that rapid evolution occurred between the genes coding for PIII-SVMPs resulting, in the production of RGD disintegrin coding genes. However, once these genes diverged, mutations in the PIII-SVMP coding genes were accumulated less frequently. PMID- 17112686 TI - Molecular analysis of Clostridium difficile isolates recovered from horses with diarrhea. AB - Clostridium difficile is an important cause of diarrhea in horses, causing sporadic and epidemic disease of varying severity. This study evaluated the molecular characteristics of 48 C. difficile isolates recovered from diarrheic horses admitted to a veterinary hospital by using PCR-ribotyping and toxin gene profile. Additionally, feces were tested for the presence of C. difficile toxin A/B via enzyme immunosorbant assay (EIA) in 38 horses. The toxin genes tcdA, tcdB and cdtB were present in 27 (56.25%), 35 (72.91%) and 2 (4.1%) strains, respectively. Eight isolates (16.6%) were A(-)B(+) variants. Thirteen of forty eight isolates (27.0%) did not posses any toxin genes (A(-)B(-)CDT(-)). A positive EIA result was reported in 17 (44%) of the cases. There was no association between the presence of different ribotypes or strains and toxin gene(s) profiles and the clinical outcome. PMID- 17112687 TI - Comparison of sensitivity and specificity in three commercial foot-and-mouth disease virus non-structural protein ELISA kits with swine sera in Taiwan. AB - Three commercialized ELISA kits for the detection of antibodies to the non structural proteins (NSPs) of FMD virus were compared, using sera from uninfected, vaccinated, challenged and naturally infected pigs. The kinetics of the antibody response to NSPs was compared on sequential serum samples in swine from challenge studies and outbreaks. The results showed that ELISA A (UBI) and ELISA B (CEDI) had better sensitivity than that of the 3ABC recombinant protein based ELISA C (Chekit). The peak for detection of antibodies to NSPs in ELISA C was significantly delayed in sera from natural infection and challenged swine as compared to the ELISA A and B. The sensitivity of the three ELISAs gradually declined during the 6-month post-infection as antibodies to NSP decline. ELISA kits A and B detected NSP antibody in 50% of challenged pigs by the 9-10th-day and 7-8th-day post-challenge, respectively. ELISA B and C had better specificity than ELISA A on sequential serum samples obtained from swine immunized with a type O FMD vaccine commercially available in Taiwan. Antibody to NSPs before vaccination was not detected in swine not exposed to FMD virus, however, antibody to NSPs was found in sera of some pigs after vaccination. All assays had significantly lower specificity when testing sera from repeatedly vaccinated sows and finishers in 1997 that were tested after the 1997 FMD outbreak. However, when testing sera from repeatedly vaccinated sows or finishers in 2003-2004, the specificity for ELISAs A, B and C were significantly better than those in 1997. This effect was less marked for ELISA A. The ELISA B was the best test in terms of the highest sensitivity and specificity and the lowest reactivity with residual NSP in vaccinates. PMID- 17112688 TI - Persistence of viral RNA in rabbits which overcome an experimental RHDV infection detected by a highly sensitive multiplex real-time RT-PCR. AB - An internally controlled multiplex real-time RT-PCR using TaqMan probes and external standards for absolute RNA quantification was developed as a new diagnostic tool for the detection of rabbit haemorrhagic disease virus (RHDV). The test revealed a specificity of 100%, an analytical sensitivity of 10 copies/well and a linearity over a range from 10(1) to 10(10) copies. The viral loads in organs, leukocytes, sera and excretions of seropositive, convalescent rabbits which were overcoming an experimental infection with RHDV were determined using the validated assay. As a result, viral RNA was demonstrated and quantified for at least 15 weeks. Thus, a persistence of viral RNA after experimental infection of rabbits could be shown for the first time. In contrast, neither antigen nor infectious virus could be detected by antigen-ELISA, immunohistochemistry or experimental transmission. Therefore, further experiments are necessary to prove that the persistence of RNA is linked with the persistence of infectious virus particles. PMID- 17112689 TI - Mothers' perception of Newborn Individualized Developmental Care and Assessment Program (NIDCAP) as compared to conventional care. AB - BACKGROUND: Family-centred care according to the Newborn Individualized Developmental Care and Assessment Program (NIDCAP) has been reported to positively influence family function. The aim was to examine if NIDCAP affects the views of prematurely born infants' mothers regarding maternal role, perception of the infant and the neonatal care. METHOD: Preterm infants with gestational age <32 weeks were randomly assigned to receive either care based on NIDCAP (n=12) or conventional neonatal care (n=13), forming two comparable groups with respect to gestational age, birth weight, female/male ratio, and initial illness severity. A questionnaire was designed to evaluate various aspects of the mothers' attitudes and apprehension of their maternal role, perception of their infant and the neonatal care. The questionnaire was validated and given to the mothers when the infants reached 36 weeks postmenstrual age (PMA). RESULTS: Ten mothers in each group replied to the questionnaire. The mothers in the NIDCAP group perceived more closeness to their infants than did the control mothers (p=0.022) and this feeling demonstrated no significant correlation to the infant's gestational age, weight at birth or severity of illness. Furthermore, the mothers in the NIDCAP-group tended to rate the staff's ability to support them in their role as a mother somewhat higher (p=0.066), but at the same time they expressed more anxiety than did the control mothers (p=0.033). CONCLUSION: Early intervention according to NIDCAP seems to facilitate a feeling of closeness between the mother and her premature infant regardless of the infant's birth weight or health status. The higher level of anxiety in the mothers in the NIDCAP group, may mirror that the mothers in the NIDCAP-group had already bonded to their infants during the hospital stay. PMID- 17112691 TI - Serum dioxin-like compounds and aromatase (CYP19) expression in endometriotic tissues. AB - Dioxin-like compounds (DLCs) are suspected etiological factors of endometriosis but their potential mechanisms of action remain elusive. Because endometriosis is an estrogen-dependent disease and since aromatase (CYP19), a key enzyme in estrogen biosynthesis, was recently demonstrated to be expressed in endometriotic lesions, we hypothesized that dioxin-like compounds could modulate local estrogen production through an up-regulation of aromatase. We tested this hypothesis by examining the correlation between serum DLC levels and CYP19 expression in endometriotic tissue obtained from 47 patients with peritoneal, ovarian endometriosis and/or deep endometriotic nodules of the rectovaginal septum. Aromatase expression was assessed by real-time RT-PCR in biopsied endometriotic tissues [peritoneal (n=19), ovarian (n=17) endometriosis and deep endometriotic nodules of the recto-vaginal septum (n=29)]. The relationship between aromatase expression and DLCs was traced by simple regression analysis. DLCs did not appear to be significant determinants of aromatase expression. CYP1A1 expression, measured as a positive control, was found associated with current smoking but not with DLCs. We conclude that DLCs do probably not facilitate the growth of endometriotic lesions by up-regulating the local expression of aromatase. PMID- 17112692 TI - Curcumin-phospholipid complex: Preparation, therapeutic evaluation and pharmacokinetic study in rats. AB - A novel formulation of curcumin in combination with the phospholipids was developed to overcome the limitation of absorption and to investigate the protective effect of curcumin-phospholipid complex on carbon tetrachloride induced acute liver damage in rats. The antioxidant activity of curcumin phospholipid complex (equivalent of curcumin 100 and 200 mg/kg body weight) and free curcumin (100 and 200 mg/kg body weight) was evaluated by measuring various enzymes in oxidative stress condition. Curcumin-phospholipid complex significantly protected the liver by restoring the enzyme levels of liver glutathione system and that of superoxide dismutase, catalase and thiobarbituric acid reactive substances with respect to carbon tetrachloride treated group (P < 0.05 and <0.01). The complex provided better protection to rat liver than free curcumin at same doses. Serum concentration of curcumin obtained from the complex (equivalent to 1.0 g/kg of curcumin) was higher (Cmax 1.2 microg/ml) than pure curcumin (1.0 g/kg) (Cmax 0.5 microg/ml) and the complex maintained effective concentration of curcumin for a longer period of time in rat serum. The result proved that curcumin-phospholipid complex has better hepatoprotective activity, owe to its superior antioxidant property, than free curcumin at the same dose level. PMID- 17112693 TI - Effect of a Psidii guajavae folium extract in the treatment of primary dysmenorrhea: a randomized clinical trial. AB - OBJECTIVE: To assess the efficacy of two different doses of a Psidii guajavae folium extract in the management of primary dysmenorrhea. METHODOLOGY: A double blinded randomized clinical trial was conducted in 197 women with primary dysmenorrhea. Four intervention groups were defined: two extract doses (3 and 6 mg/day); ibuprofen (1200 mg/day); placebo (3mg/day). Participants were followed up individually for 4 months. The main outcome variable was abdominal pain intensity measured according to a visual analogue scale (VAS). RESULTS: The average age of participants was 19 years; menarche occurred around age 12 years. Participants had menstrual cycles of 28 or 29 days, with menstruation lasting 5 days and mean of pain intensity of 8.2 on the VAS. During each successive treatment cycle, participants experienced a lower pain intensity score. Multiple regression analysis, after adjusting each cycle for baseline pain, treatment compliance and other variables, showed that the group receiving 6 mg/day extract had significantly reduced pain intensity (p<0.001). This effect was maintained in cycles 2 and 3, although the reduction in the mean of pain intensity was lower. The group receiving the 3mg/day extract did not show a consistent effect throughout the three cycles. CONCLUSION: At a dose of 6 mg/day, the standardized phyto-drug (Psidii guajavae folium extract) reduced menstrual pain significantly compared with conventional treatment and placebo. PMID- 17112694 TI - Phenolic-rich fraction from Rhus verniciflua Stokes (RVS) suppress inflammatory response via NF-kappaB and JNK pathway in lipopolysaccharide-induced RAW 264.7 macrophages. AB - The effects of phenolic-rich fraction (PRF) from Rhus verniciflua Stokes (Anacardiaceae) on the activities of cellular signaling molecules that mediate inflammatory responses in LPS-induced RAW 264.7 macrophages were investigated. At various concentrations of PRF significantly inhibited NO, PGE(2) and TNF-alpha production in LPS-induced RAW 264.7 macrophage cells. The PRF also significantly inhibited iNOS and COX-2 protein expression in LPS-induced RAW 264.7 macrophage in a concentration-dependent manner. Transcription factor NF-kappaB plays a key role for the inducible expression of genes mediating proinflammatory effects and here, we show that PRF can inhibit the induction of NF-kappaB activity. The PRF effectively inhibited the iNOS and COX-2 protein expression through suppression of phospho-JNK1/2 activation. Study using PDA HPLC has found that the PRF contains several low molecular compounds (i.e. p-coumaric acid, fustin, kaempferol-3-O-glucoside, sulfuretin, butein, kaempferol). Our results indicate that the anti-inflammatory properties of PRF might result from the inhibition of pro-inflammatory mediators (e.g., NO, PGE(2) and TNF-alpha) by suppression of such signaling pathways as NF-kappaB and JNK1/2. PMID- 17112695 TI - Inhibition of the pro-inflammatory mediators' production and anti-inflammatory effect of the iridoid scrovalentinoside. AB - We have studied scrovalentinoside, an iridoid with anti-inflammatory properties isolated from Scrophularia auriculata ssp. pseudoauriculata, as an anti inflammatory agent in different experimental models of delayed-type hypersensitivity. We found that scrovalentinoside reduced the edema induced by oxazolone at 0.5 mg/ear and sheep red blood cells at 10 mg/kg. The observed effect occurred during the last phase or inflammatory response; during the earlier phase or induction of the delayed-type hypersensitivity reaction, no significant activity was noted. Thus, scrovalentinoside reduced both the edema and cell infiltration in vivo and reduced lymphocyte proliferation in vitro, affecting the cycle principally during the first 48 h. Whereas cells stimulated with phytohemagglutinin changed from the G(0)/G(1) phase to the S and G(2)/M phases, when these same cells were treated with scrovalentinoside (100 microM), they remained in the G(0)/G(1) phase. Finally, scrovalentinoside inhibited the production of the pro-inflammatory mediators' TNF-alpha, IFN-gamma, IL-1beta, IL 2, IL-4, LTB(4), and NO, but had no effect on the production of the anti inflammatory cytokine IL-10. PMID- 17112696 TI - Significance of mesenteric lymphadenopathy after pancreaticoduodenectomy for periampullary carcinomas: evaluation with serial MDCT studies. AB - OBJECTIVE: To evaluate the significance of mesenteric lymphadenopathy arising after pancreaticoduodenectomy (PD) for periampullary carcinomas. MATERIALS AND METHODS: Serial multidetector CT (MDCT) studies in 44 patients after PD for periampullary carcinomas (Group A) were retrospectively reviewed, the incidence and interval changes of mesenteric lymphadenopathy were evaluated, and comparisons were made with 21 patients who underwent PD for benign diseases (Group B). RESULTS: In Group A, mesenteric lymphadenopathy was seen in 31 of 44 (70.5%) patients. The short axis diameter of the largest lymph node ranged from 6.2 to 11.5mm (mean+/-S.D.: 8.6+/-1.4mm), and the short-axis-to-long-axis (S/L) ratio ranged from 0.46 to 0.99 (mean+/-S.D.: 0.69+/-0.12). Six of the 31 (19.4%) mesenteric lymphadenopathy cases showed interval increases in size on follow-up studies, and three cases showed new development of mesenteric lymphadenopathy: all of these nine cases had concurrent recurrence in the mesenteric root. In Group B, 11 (52.4%) had mesenteric lymphadenopathy. The short axis diameter of the largest lymph node ranged from 5.6 to 10.9mm (mean+/-S.D.: 8.0+/-1.9mm) and the S/L ratio ranged from 0.48 to 0.93 (mean+/-S.D.: 0.67+/-0.12). Differences in the incidence, short axis diameter and S/L ratio were not significant between Groups A and B (p=0.154, 0.271 and 0.654, respectively). CONCLUSION: Mesenetric lymphadenopathy after PD less likely reflects recurrence. Lymph node metastasis may be suggested only when coexisting recurrent mass is found in the proximal mesenteric root. PMID- 17112697 TI - Repeat liver resection for recurrent liver metastases from colorectal cancer. AB - AIMS: Numerous patients suffer from recurrence after resection of liver metastases from colorectal cancer. Recurrence is frequently restricted to the liver and repeat liver resection may offer a curative option in these patients. This study was conducted to clarify safety and effectiveness of this treatment and to identify prognostic factors of a favourable outcome after repeat hepatectomy. METHODS: Between January 1988 and March 2006 in our institution 811 patients underwent 841 liver resections for metastases from colorectal cancer. Among these, 94 patients underwent a repeat hepatectomy. Patients were identified from a prospective database and retrospectively reviewed. Results of different time periods were assessed and prognostic factors for a favourable outcome were determined. RESULTS: The perioperative morbidity and mortality was 24% (23 of 94) and 3% (3 of 94), respectively. The one-, three-, five- and ten-year survival for all patients in this series was 89%, 55%, 38% and 23%, respectively. In the univariate analysis, pT-stage of the primary, diameter of the largest metastases, surgical radicality, period of resection and distribution of metastases showed statistically significant influence on survival. The multivariate analysis revealed only pT-stage of the primary tumour, surgical radicality and period of resection as independent prognostic factors. CONCLUSIONS: Repeat hepatectomy is a safe and effective treatment for recurrent liver metastases from colorectal cancer. Perioperative risk and long-term survival were similar when compared to the results obtained during the initial resection. Achieving a curative resection is the most relevant prognostic factor for a favourable prognosis after repeat liver resection. PMID- 17112698 TI - A 7 year experience with immediate breast reconstruction after skin sparing mastectomy for cancer. AB - Skin sparing mastectomy (SSM) is a procedure that has gained popularity for patients who elect to have a mastectomy as the treatment for carcinoma of the breast. Reconstruction of the breast after SSM yields the best aesthetic results since it preserves most of the skin envelope and the inframammary fold. The purpose of this study was to evaluate the oncologic safety and aesthetic results for SSM and immediate breast reconstruction. One hundred and five patients treated between April 1997 and December 2004 were reviewed. The mean age of the patients was 40 years. The mean follow up was for 48 months, considered from the time of SSM and reconstruction. Reconstructive techniques included pedicled transverse rectus abdominis muscle (TRAM) flap (n: 70 patients), tissue expanders and implants (n: 29 patients), latissimus dorsi flaps and implants (n: 6 patients). Aesthetic results were judged by an independent plastic surgeon and by the patient as excellent, good, fair or poor. The findings of this study demonstrate that SSM and immediate breast reconstruction can be safely performed. Detection and treatment of recurrences were not inhibited by the reconstructions. In this experience the best results were seen after TRAM flap reconstructions. The general level of satisfaction expressed by the patient was correlated with a good appearance of the reconstructed breast and the physical comfort. PMID- 17112699 TI - Analysis of the genomic structure of the porcine CD1 gene cluster. AB - CD1 is an MHC class I-like protein that presents lipid antigens to T cell receptors. We determined 470,187 bp of the genomic sequence encompassing the region encoding porcine CD1 genes. We identified 16 genes in this region and newly identified CD1A2, CD1B, CD1C, CD1D, and CD1E. Porcine CD1 genes were located in clusters between KIRREL and olfactory receptor (OR) genes, as observed in humans, although they were divided into two regions by a region encoding OR genes. Comparison of the genomic sequences of CD1 gene loci in pigs with other mammals showed that separation of the CD1 gene cluster by ORs was observed only in pigs. CD1A duplication in the porcine genome was estimated to have occurred after the divergence of the human and porcine. This analysis of the genomic sequence of the porcine CD1 family will contribute to our understanding of the evolution of mammalian CD1 genes. PMID- 17112701 TI - BMP2 and BMP6 control p57(Kip2) expression and cell growth arrest/terminal differentiation in normal primary human epidermal keratinocytes. AB - Functional studies of the canonical Bone Morphogenetic Protein (BMP) signalling pathway in human epidermal keratinocytes have been limited to the immortalized and p53-mutated HaCaT cells and are primarily dependent on BMP6 treatment in mouse epidermal keratinocytes. Despite these insightful analyses, the molecular mechanism underlying the role of BMP signalling in the precise balance between growth arrest and terminal differentiation of keratinocytes still remains not clearly defined. The current study first investigated the hitherto uncharacterized status and functions of BMP signalling in normal human keratinocytes by using three independent strains of primary interfollicular epidermal keratinocytes. Then we provided data demonstrating the role of BMP2 compared to BMP6 in the inhibition of growth and induction of subsequent terminal differentiation of these cells. A second relevant finding is based on the clonal analysis of colony types present in untreated and BMP2/6-treated cultures in absence of EGF. BMP treatment results in the clonal transition from proliferative to abortive colonies, suggesting that BMP signalling most likely inhibits stem cell proliferation and triggers cell cycle exit from transit amplifying cells. Third, we showed evidence that, of the three members of the Cip/Kip family of cyclin-dependent kinase inhibitors, only p57(Kip2) and p21(Cip1) have a BMP2/6 induced expression. One mechanism of inhibition of cell proliferation involves p57(Kip2) as an immediate early response, in contradistinction with p21(Cip1) which largely depends on de novo protein synthesis for its effect to proceed. All together, these results clarify the BMP signalling status in normal primary human keratinocytes and support a new mechanism of inhibition of the proliferation of interfollicular epidermal keratinocytes coupled with induction of their terminal differentiation following BMP2 or BMP6 addition. PMID- 17112700 TI - Gestational toluene exposure effects on spontaneous and amphetamine-induced locomotor behavior in rats. AB - Gestational Toluene Exposure Effects on Spontaneous and Amphetamine-Induced Locomotor Behavior in Rats. Bowen, S.E., Mohammadi, M.H., Batis, J.C., and Hannigan, J.H. Neurotoxicology and Teratology, XX, 2006. The abuse of volatile organic solvents (inhalants) continues to be a major health concern throughout the world. Toluene, which is found in many products such as glues and household cleaners, is among the most commonly abused organic solvents. The neurobehavioral teratogenic sequelae of solvent abuse (i.e., repeated, brief inhalation exposures to very high concentrations of solvents) have not been examined thoroughly. In a preclinical model of inhalant abuse, timed-pregnant Sprague-Dawley rats were exposed to 0, 8000, or 12,000 parts per million (ppm) for 15 min twice daily from gestation day 8 (GD8) through GD20. In the first experiment, separate groups of offspring were observed individually in an open-field on postnatal day 22 (PN22), PN42 or PN63. In the second experiment, other offspring given identical prenatal toluene exposures were observed in an "open-field" following an acute i.p. injection of amphetamine (0, 0.56, 1.78 mg/kg) on PN28. Automated measurements of distance traveled and ambulatory time were recorded. Prenatal toluene exposure resulted in small alterations in spontaneous activity compared to non-exposed rats. Prenatal exposure to 12,000 ppm toluene resulted in significant hyposensitivity to the locomotor stimulatory effects of the amphetamine challenge in male but not female rats on PN28. The results demonstrate that prenatal exposure to abuse patterns of high concentrations of toluene through inhalation can alter spontaneous and amphetamine-induced locomotor behavior in rats. The expression of these effects also appears to depend upon the postnatal age of testing. These results imply that abuse of organic solvents during pregnancy in humans may also produce long-lasting effects on biobehavioral development. PMID- 17112702 TI - Protective effect of alpha-lipoic acid in experimental spermatic cord torsion. AB - OBJECTIVE: We investigated the effect of alpha-lipoic acid (LA) in rats that were subjected to torsion/detorsion of the spermatic cord in a comparative controlled experiment. METHODS: Forty-eight male Wistar rats, randomized in two groups, received intraperitoneal injections of LA (LA group; aqueous solution at 36 mg/kg of body weight per day) or equal volume of saline (control group) 21, 9, and 1 h before torsion of the spermatic cord. Rats in each group were distributed in four subgroups, each comprising six animals. All surgical procedures were performed under inhaled ether anesthesia. Ischemia was induced by 720-degree torsion of the spermatic cord for 3 h. The right testis was assessed through longitudinal scrotal incision. After each surgical procedure, scrotal incisions were closed with 4-0 nylon monofilament. Ipsilateral testes and arterial blood samples were collected at the end of ischemia and 1, 3, and 6 h after detorsion. Thiobarbituric acid-reactive substances and reduced glutathione levels (micromoles per gram of wet tissue) were assayed in testis. Total antioxidant power was measured in blood plasma. RESULTS: LA pretreatment promoted a significant decrease in testicular concentrations of thiobarbituric acid-reactive substances and simultaneously induced an increase in reduced glutathione concentrations at all time points studied. Plasma total antioxidant power levels increased significantly during reperfusion (T-1) in LA-treated rats compared with control rats. CONCLUSION: LA administered before torsion of the spermatic cord showed significant protective effects against ischemia/reperfusion injury by decreasing lipid peroxidation and regulating testicular reduced glutathione and plasma total antioxidant power levels. PMID- 17112703 TI - Analgesic and anti-inflammatory dose-response relationship of 7.5 and 15 mg meloxicam after lower third molar removal: a double-blind, randomized, crossover study. AB - Fifty patients were scheduled to undergo removal of symmetrically positioned lower third molars in two separate appointments. Meloxicam 7.5 or 15 mg was once daily administered in a double-blind, randomized and crossover manner after the surgery for 4 days. Objective and subjective parameters were recorded for comparison of postoperative courses. Patients treated with 7.5mg meloxicam who underwent osteotomy reported higher pain scores at 1.5, 3, 4, 10, 12 and 16 h (P<0.05) and ingested a greater amount of rescue analgesic medication (P<0.05) than those who did not require osteotomy. A higher percentage of patients who underwent osteotomy medicated with 7.5mg meloxicam needed rescue medication as compared to those who did not require osteotomy (P<0.05). There was a similar mouth opening at suture removal compared with preoperative values for both doses (P>0.05). There were no significant differences concerning swelling observed on the 2nd or 7th postoperative days in comparison with baseline (P>0.05) between the two doses. Pain, trismus and swelling after lower third molar removal not requiring osteotomy can be successfully controlled by a dose regimen of 7.5mg meloxicam once daily. For more aggressive extractions 15 mg meloxicam is advisable. PMID- 17112704 TI - Povidone iodine as a haemostatic agent. PMID- 17112705 TI - Ultrasonic bone cutting for surgically assisted rapid maxillary expansion (SARME) under local anaesthesia. AB - Ultrasonic bone-cutting surgery has been recently introduced as a feasible alternative to the conventional tools of cranio-maxillo-facial surgery, due to its technical characteristics of precision and safety. The device used is unique in that the cutting action occurs when the tool is employed on mineralized tissues, but stops on soft tissues. This technical note illustrates the use of Piezosurgery for all osteotomies of surgically assisted rapid maxillary expansion (SARME). The procedure, including pterygo-maxillary detachment, can be completed under local anaesthesia. Other advantages include minimal risk of jeopardizing critical anatomic structures (e.g. palatine artery), minimal intraoperative bleeding and postoperative swelling, and minimal thermal damage to bone surfaces. Narrow and rectilinear osteotomies can be easily performed with varying vibrating scalpels, at the cost of a longer operative time. PMID- 17112706 TI - Occurrence of plasmidic AmpC type beta-lactamase-mediated resistance in Escherichia coli: report from the SENTRY Antimicrobial Surveillance Program (North America, 2004). AB - Among 1429 Escherichia coli isolates collected as part of the SENTRY Antimicrobial Surveillance Program (2004) from 30 North American medical centres, 65 (4.5%) were screen-positive for an extended-spectrum beta-lactamase (ESBL). Among the strains with a negative ESBL confirmatory test (n=26; clavulanic acid inhibition), a CMY-2 enzyme was detected in 13 isolates (50.0%), FOX-5 in 3 isolates (11.5%) and DHA-1 in 1 isolate (3.8%). These AmpC-producing E. coli were cephamycin (cefoxitin)-resistant but susceptible to cefepime (minimum inhibitory concentration (MIC) < OR =0.12-4 mg/L). Clearly, the ESBL tests recommended by the Clinical and Laboratory Standards Institute identify only a fraction of E. coli with elevated beta-lactam MIC values as ESBL-producing strains; the majority of the remaining strains would be potentially responsive to some other beta lactams, directed by accurately performed and interpreted susceptibility methods. PMID- 17112707 TI - BMP signaling regulates PGC numbers and motility in organ culture. AB - Members of the bone morphogenetic protein (BMP) family play diverse roles in multiple developmental processes. However, in the mouse, mutations in many BMPs, BMP receptors and signaling components result in early embryonic lethality making it difficult to analyze the role of these factors during organogenesis or tissue homeostasis in the adult. To bypass this early lethality, we used an organ culture system to study the role of BMPs during primordial germ cell (PGC) migration. PGCs are the embryonic precursors of the sperm and eggs. BMPs induce formation of primordial germ cells within the proximal epiblast of embryonic day 7.5 (E7.5) mouse embryos. PGCs then migrate via the gut to arrive at the developing gonads by E10.5. Addition of BMP4 or the BMP-antagonist Noggin to transverse slices dissected from E9.5 embryos elevated PGC numbers or reduced PGC numbers, respectively. Noggin treatment also slowed and randomized PGC movements, resulting in a failure of PGCs to colonize the urogenital ridges (UGRs). Based on p-Smad1/5/8 staining, migratory PGCs do not respond to endogenous BMPs. Instead, the somatic cells of the urogenital ridges exhibit elevated p-Smad1/5/8 staining revealing active BMP signaling within the UGRs. Noggin treatment abrogated p-Smad staining within the UGRs and blocked localized expression of Kitl, a cytokine known to regulate the survival and motility of PGCs and Id1, a transcription factor expressed within the UGRs. We propose that BMP signaling regulates PGC migration by controlling gene expression within the somatic cells along the migration route and within the genital ridges. PMID- 17112708 TI - Protein adsorption and stability of poly(ethylene oxide)-modified surfaces having hydrophobic layer between substrate and polymer. AB - The materials covered with poly(ethylene oxide) (PEO) are of use in a wide variety of biomaterials due to blood compatibility of this polymer. The long-term sustainability of its blood compatibility strongly depends on the stability of the PEO layer against aqueous environment. An attempt was made in the present work to immobilize a PEO layer on the silicon surfaces using a silane coupling agent with the aim to improve the waterproof durability of the layer. Several kinds of PEO-modified substrates having a densely and closely packed hydrocarbon layer between substrate and PEO layer were prepared and the stability of the PEO layer against phosphate buffer saline (pH 7.4) was examined in terms of the density of hydrocarbon chains. Those substrates which have a dense hydrophobic chain layer showed a high waterproof durability and a good ability to suppress protein adsorption. PMID- 17112709 TI - Influence of the anisotropic polarizability on the anomalous dielectric relaxation spectra. AB - The nonlinear dielectric response due to the application of a strong dc bias electric field superimposed on a weak ac electric field is considered in the context of the anomalous diffusion (subdiffusion). A perturbation procedure is used to derive analytical expressions for the first three harmonic components of the electric polarization of an assembly of both polar and anisotropically polarizable symmetric-top molecules. To accomplish that, an infinite hierarchy of multiterm (21) differential-recurrence equations of noninteger order for the moments is established and solved for the stationary regime. The results so obtained are illustrated in the form of Argand diagrams and three-dimensional relaxation spectra for the complex nonlinear dielectric increment extracted from the first harmonic component of the electric susceptibility. These plots show the role and importance played by the fractional exponent alpha and the parameter P measuring the influence of the dipole moment over the permanent one. PMID- 17112710 TI - Surface modification on microfluidic devices with 2-methacryloyloxyethyl phosphorylcholine polymers for reducing unfavorable protein adsorption. AB - Surface modification of polymer materials for preparing microfluidic devices including poly(dimethyl siloxane) (PDMS) was investigated with phospholipids polymers such as poly(2-methacryloyloxylethyl phosphorylcholine(MPC)-co-n-butyl methacrylate) (PMB) and poly(MPC-co-2-ethylhexyl methacrylate-co-2-(N,N dimethylamino)ethyl methacrylate) (PMED). The hydrophilicity of every surface on the polymer materials modified with these MPC polymers increased and the value of zeta-potential became close to zero. The protein adsorption on the polymer materials with and without the surface modification was evaluated using a protein mixture of human plasma fibrinogen and serum albumin. Amount of proteins adsorbed on these polymeric materials showed significant reduction by the surface modification with the MPC polymers compared to the uncoated surfaces ranging from 56 to 90%. Furthermore, we successfully prepared PDMS-based microchannel which was modified by simple coating with the PMB and PMED. The modified microchannel also revealed a significant reduction of adsorption of serum albumin. We conclude that the MPC polymers are useful for reducing unfavorable protein adsorption on microfluidic devices. PMID- 17112711 TI - Light scattering study of complex formation between protein and polyelectrolyte at various ionic strengths. AB - Formation of protein-polyelectrolyte complexes (PPCs) between bovine serum albumin (BSA) and potassium poly (vinyl alcohol) sulfate (KPVS) was studied at pH 3 as a function of ionic strength. Turbidimetric titration was employed by a combination of dynamic light scattering (DLS) and electrophoretic light scattering (ELS). The formal charge (Z(PPC)) of the resulting PPCs at different ionic strengths were estimated from ELS data by assuming the free draining and the non-free draining model. The radius of a BSA molecule in the complex was used in the former model for calculation of Z(PPC) with the Henry's equation, while in the latter case the hydrodynamic radius of a PPC particle determined from DLS was employed. The results obtained were compared with the Z(PPC) values calculated using a relation of Z(PPC)=n(b)Z(BSA)+alphaZ(KPVS), where Z(BSA) (> or =0) and Z(KPVS) (< or =0) denote the formal charge of BSA and KPVS, respectively. Moreover, n(b) is the number of bound proteins per complex composed of alpha polymer chains. It was suggested that the PPC between BSA and KPVS behaves as a free draining molecule during the electrophoresis, at least at a high ionic strength. Also suggested is that the PPC formation at low ionic strength follows a 1:1 stoichiometry in the charge neutralization. PMID- 17112712 TI - Identification and differentiation of Lactobacillus, Streptococcus and Bifidobacterium species in fermented milk products with bifidobacteria. AB - The aim of this study was to identify and discriminate bacteria contained in commercial fermented milks with bifidobacteria by the use of amplified ribosomal DNA restriction analysis (ARDRA) and randomly amplified polymorphic DNA (RAPD) techniques. ARDRA of the 16S rDNA gene and RAPD were performed on 13 Lactobacillus strains, 13 Streptococcus and 13 Bifidobacterium strains isolated from commercial fermented milk. Lactobacillus delbrueckii, Streptococcus thermophilus and Bifidobacterium animalis isolates were identified by genus- and species-PCR and also, they were differentiated at genus and species level by ARDRA using MwoI restriction enzyme. The ARDRA technique allowed for the discrimination among these three related genus with the use of only one restriction enzyme, since distinctive profiles were obtained for each genus. Therefore it can be a simple, rapid and useful method for routine identification. Also, RAPD technique allowed the discrimination of all bacteria contained in dairy products, at genus- and strain-level by the performance of one PCR reaction. PMID- 17112713 TI - Collagen IX is indispensable for timely maturation of cartilage during fracture repair in mice. AB - Fracture repair recapitulates in adult organisms the sequence of cell biological events of endochondral ossification during skeletal development and growth. After initial inflammation and deposition of granulation tissue, a cartilaginous callus is formed which, subsequently, is remodeled into bone. In part, bone formation is influenced also by the properties of the extracellular matrix of the cartilaginous callus. Deletion of individual macromolecular components can alter extracellular matrix suprastructures, and hence stability and organization of mesenchymal tissues. Here, we took advantage of the collagen IX knockout mouse model to better understand the role of this collagen for organization, differentiation and maturation of a cartilaginous template during formation of new bone. Although a seemingly crucial component of cartilage fibrils is missing, collagen IX-deficient mice develop normally, but are predisposed to premature joint cartilage degeneration. However, we show here that lack of collagen IX alters the time course of callus differentiation during bone fracture healing. The maturation of cartilage matrix was delayed in collagen IX-deficient mice calli as judged by collagen X expression during the repair phase and the total amount of cartilage matrix was reduced. Entering the remodeling phase of fracture healing, Col9a1(-/-) calli retained a larger percentage of cartilage matrix than in wild type indicating also a delayed formation of new bone. We concluded that endochondral bone formation can occur in collagen IX knockout mice but is impaired under conditions of stress, such as the repair of an unfixed fractured long bone. PMID- 17112714 TI - Differential expression of two tropoelastin genes in zebrafish. AB - Elastin is the extracellular matrix protein responsible for properties of extensibility and elastic recoil in large blood vessels, lung and skin of most vertebrates. Elastin is synthesized as a monomer, tropoelastin, but is rapidly transformed into its final polymeric form in the extracellular matrix. Until recently information on sequence and developmental expression of tropoelastins was limited to mammalian and avian species. We have recently identified and characterized two expressed tropoelastin genes in zebrafish. This was the first example of a species with multiple tropoelastin genes, raising the possibility of differential expression and function of these tropoelastins in elastic tissues of the zebrafish. Here we have investigated the temporal expression and tissue distribution of the two tropoelastin genes in developing and adult zebrafish. Expression was detected early in skeletal cartilage structures of the head, in the developing outflow tract of the heart, including the bulbus arteriosus and the ventral aorta, and in the wall of the swim bladder. While the temporal pattern of expression was similar for both genes, the upregulation of eln2 was much stronger than that of eln1. In general, both genes were expressed and their gene products deposited in most of the elastic tissues examined, with the notable exception of the bulbus arteriosus in which eln2 expression and its gene product was predominant. This finding may represent a sub-specialization of eln2 to provide the unique architecture of elastin and the specific mechanical properties required by this organ. PMID- 17112715 TI - Exacerbations and lung function decline in COPD: new insights in current and ex smokers. AB - AIM: To investigate whether there is a significant relationship between an increased frequency of exacerbations and the rate of forced expiratory volume in 1s (FEV(1)) decline in COPD patients. METHODS-MEASUREMENTS: About 102 COPD patients (44 smokers, 58 ex-smokers) participated in a 3-year prospective study. Exacerbations were identified as worsening of patient's respiratory symptoms as recorded on diary cards. Spirometry was performed every 6 months. The effect of frequent exacerbations on lung function was investigated using random effects models. RESULTS: The median (mean(95% CI)) annual exacerbation rate was 2.85 (3.1 (2.7-3.6)). Patients with an annual exacerbation rate over the median rate had significantly lower baseline post-bronchodilation FEV(1)(%pred), higher MRC dyspnoea score and chronic cough compared to patients who had an annual exacerbation rate less than the median. The average annual rate of FEV(1)(%pred), adjusted for smoking decline (DeltaFEV(1)), was found significantly increased in frequent compared to infrequent exacerbators (P=0.017). The highest DeltaFEV(1) was observed in smokers frequent exacerbators and a significant interaction between exacerbation frequency and DeltaFEV(1) was also observed in ex-smokers. CONCLUSIONS: Our findings suggest that an increased frequency of exacerbations is significantly associated with FEV(1) decline even in ex-smokers. Thus, smoking and frequent exacerbations may have both negative impact on lung function. Smoking cessation and prevention of exacerbations should be a major target in COPD. PMID- 17112716 TI - Life cycle assessment of the waste hierarchy--a Danish case study on waste paper. AB - The waste hierarchy is being widely discussed these days, not only by cost benefit analysts, but a growing number of life cycle assessments (LCA) have also begun to question it. In this article, we investigate the handling of waste paper in Denmark and compare the present situation with scenarios of more waste being recycled, incinerated or consigned to landfill. The investigations are made in accordance with ISO 14040-43 and based on the newly launched methodology of consequential LCA and following the recent guidelines of the European Centre on Waste and Material Flows. The LCA concerns the Danish consumption of paper in 1999, totalling 1.2 million tons. The results of the investigation indicate that the waste hierarchy is reliable; from an environmental point of view recycling of paper is better than incineration and landfilling. For incineration, the reason for the advantage of landfilling mainly comes from the substitution of fossil fuels, when incinerators provide heat and electricity. For recycling, the advantage is related to the saved wood resources, which can be used for generating energy from wood, i.e., from renewable fuel which does not contribute to global warming. PMID- 17112717 TI - Management of construction and demolition waste. PMID- 17112718 TI - Multiphase electrodes for microbead control applications: integration of DEP and electrokinetics for bio-particle positioning. AB - Advances in microfabrication have introduced new possibilities for automated, high-throughput biomedical investigations and analysis. Physical effects such as dielectrophoresis (DEP) and AC electrokinetics can be used to manipulate particles in solution to coordinate a sequence of bioanalytical processing steps. DEP is accomplished with non-uniform electric fields that can polarize particles (microbeads, cells, viruses, DNA, proteins, etc.) in suspension causing translational or rotational movement. AC electrokinetics is another phenomena involved with movement of particles in suspension with electric fields and is comprised of both electro-thermal and electro-osmotic effects. This paper investigates single layer electrodes that are effective for particle localization and clustering based on DEP and AC electrokinetic effects. We demonstrate a novel multi-electrode setup capable of clustering particles into an array of discrete bands using activated and electrically floating electrodes. These bands shift to adjacent regions on the electrode surface by altering the electrode activation scheme. The predictability of particle placement to specific locations provides new opportunities for integration and coordination with raster scanning lasers or a charge coupled device (CCD) for advanced biomedical diagnostic devices, and more sophisticated optical interrogation techniques. PMID- 17112720 TI - Opportunities for structure-based design of protease-directed drugs. AB - As a result of the recent enormous technological progress, experimental structure determination has become an integral part of the development of drugs against disease-related target proteins. The post-translational modification of proteins is an important regulatory process in living organisms; one such example is lytic processing by peptidases. Many different peptidases represent disease targets and are being used in structure-based drug design approaches. The development of drugs such as aliskiren and tipranavir, which inhibit renin and HIV protease, respectively, testifies to the success of this approach. PMID- 17112721 TI - Degradation of cyanide by Trichoderma mutants constructed by restriction enzyme mediated integration (REMI). AB - REMI technique was used to construct mutants with improved cyanide-degradation ability from biocontrol fungus Trichoderma koningii strain T30. The plasmid pV2 transformation was confirmed by PCR and Southern blot analysis. Out of 21 transformants, 15 single-copied transformants (71.4%) were found. To compare enzyme activities of rhodanese and cyanide hydratase, T. atroviride T23, T. harzianum T21 and their transformants constructed by REMI previously were also included. Transformants TkB6 (0.173 micromols thiocyanate formed min(-1)mg protein(-1)) from T30 and TaK1 (0.174 micromols thiocyanate formed min(-1)mg protein(-1)) from T23 showed higher rhodanese activity than other transformants and their wild strains. TkA9 (5.53 micromols formamide formed h(-1)mg protein( 1)) from T30 and Th64 (5.35 micromols formamide formed h(-1)mg protein(-1)) from T21 had higher cyanide hydratase activity than other transformants and their wild strains. PMID- 17112722 TI - Solid phase-assisted synthesis and screening of a small library of N-(4 hydroxyphenyl)retinamide (4-HPR) analogs. AB - Using solid phase-assisted synthesis and purification, a 49 member library of analogs of the mammary tumor chemopreventive retinoid N-(4 hydroxyphenyl)retinamide (4-HPR) has been prepared. After prescreening for growth inhibitory activity in human mammary tumor cells (MCF-7) in culture, most of those analogs which showed activity (12 of them) were assayed for apoptosis inducing activity in the MCF-7 cells. At least 3 of the analogs (13, 24, and 28) showed activity approaching that of 4-HPR. PMID- 17112723 TI - Natural dibenzoxazepinones from leaves of Carex distachya: Structural elucidation and radical scavenging activity. AB - Two new dibenzoxazepinones have been isolated from the leaves of Carex distachya, an herbaceous plant growing in the Mediterranean area. The structures have been elucidated on the basis of their spectroscopic properties. Bidimensional NMR (DQ COSY, TOCSY, NOESY, ROESY, HSQC, and HMBC) furnished important data useful for the characterization of the molecules. The compounds have been assayed, for the antioxidant activity, by measuring its capacity to scavenge the DPPH, the superoxide anion, and nitric oxide radicals. PMID- 17112724 TI - Dopamine-selective potentiometric responses by new ditopic sensory elements based on a hexahomotrioxacalix[3]arene. AB - New ditopic sensory elements 2 and 3 for catecholamines based on a hexahomotrioxacalix[3]arene, with a boronic acid substituent appended, were designed and synthesized. As an interesting mode of molecular recognition at membrane surfaces, the host, when incorporated into poly(vinyl chloride) (PVC) liquid membranes, displayed excellent potentiometric selectivity for dopamine over other catecholamines (noradrenaline and adrenaline) and inorganic cations (Na+, K+, and NH4+). PMID- 17112725 TI - Beta-secretase (BACE-1) inhibitors: accounting for 10s loop flexibility using rigid active sites. AB - BACE-1 is a flexible enzyme with experimentally determined motion in the flap region, the catalytic aspartates, and the 10s loop. Four in-house crystallographically determined complexes of tertiary carbinamine inhibitors revealed 10s loop motion in the S(3) pocket. These X-ray structures were used to correlate K(i) values, which span over five orders of magnitude, with the calculated interaction energy, using the Merck Molecular Force Field for a series of 19 tertiary carbinamine inhibitors. PMID- 17112726 TI - Human Wapl is a cohesin-binding protein that promotes sister-chromatid resolution in mitotic prophase. AB - BACKGROUND: The linkage between duplicated chromosomes (sister chromatids) is established during S phase by the action of cohesin, a multisubunit complex conserved from yeast to humans. Most cohesin dissociates from chromosome arms when the cell enters mitotic prophase, leading to the formation of metaphase chromosomes with two cytologically discernible chromatids. This process is known as sister-chromatid resolution. Although two mitotic kinases have been implicated in this process, it remains unknown exactly how the cohesin-mediated linkage is destabilized at a mechanistic level. RESULTS: The wings apart-like (Wapl) protein was originally identified as a gene product that potentially regulates heterochromatin organization in Drosophila melanogaster. We show that the human ortholog of Wapl is a cohesin-binding protein that facilitates cohesin's timely release from chromosome arms during prophase. Depletion of Wapl from HeLa cells causes transient accumulation of prometaphase-like cells with chromosomes that display poorly resolved sister chromatids with a high level of cohesin. Reduction of cohesin relieves the Wapl-depletion phenotype, and depletion of Wapl rescues premature sister separation observed in Sgo1-depleted or Esco2-depleted cells. Conversely, overexpression of Wapl causes premature separation of sister chromatids. Wapl physically associates with cohesin in HeLa-cell nuclear extracts. Remarkably, in vitro reconstitution experiments demonstrate that Wapl forms a stoichiometric, ternary complex with two regulatory subunits of cohesin, implicating its noncatalytic function in inactivating cohesin's ability to interact with chromatin. CONCLUSIONS: Wapl is a new regulator of sister chromatid resolution and promotes release of cohesin from chromosomes by directly interacting with its regulatory subunits. PMID- 17112727 TI - Reliability of the intrinsic and extrinsic patterns of level walking in older women. AB - The aim of the study was to assess short- and long-term reliability of the harmonic analysis of the trajectories of head, upper trunk, and pelvis during walking in healthy older women, which reveals mechanisms to coordinate upper body segments and control head stability in the search for a safer gait. A stereophotogrammetric system was used to measure the displacement of markers located at head, shoulder, and pelvis level in 11 healthy older women (77+/-2 years) walking on an oval shaped 20-m walkway circuit, in three experimental sessions separated by 6 weeks. The harmonic analysis of the time-curves was highly reliable both within and between sessions (standard error of measurement ranging between 0.15 mm and 1.50mm for the amplitudes and 0.03 rad and 0.41 rad for the phases) and revealed different oscillatory patterns for the pelvis, head, and upper trunk. An intrinsic pattern, representing the natural overall movement symmetry of the whole population of older women of this study, was described by the first medio-lateral harmonic and second antero-posterior and vertical harmonics. An extrinsic pattern, characteristic of each individual in the population, was described by the first antero-posterior and vertical harmonics. The intrinsic pattern was both intra- and inter-subject repeatable (coefficient of multiple correlation, CMC, ranging between 0.82 and 0.99), while the extrinsic pattern was only intra-subject repeatable (CMC ranging between 0.70 and 0.90). Harmonic analysis reliably describes upper body kinematics in older women for detecting the intrinsic and extrinsic patterns of gait, which reveal fundamental mechanisms governing their walking. PMID- 17112728 TI - Does preserving memory correlate with surviving HIV? AB - The holy grail for HIV-1 vaccine researchers is to develop an efficacious vaccine, a goal that would be aided by defining a correlate of protection from infection. Recently, the authors of two papers have come one step closer to this goal with the definition of a correlate of survival following simian immunodeficiency virus (SIV) infection: the preservation of memory CD4(+) T cells during acute infection. Both of these research groups show that vaccination can prevent the initial immune devastation caused by SIV infection and that this correlates with survival following SIV challenge. Here, we highlight the significance of these two recent papers in light of the urgent need to produce an efficacious HIV vaccine and discuss several crucial issues that must be addressed before the correlate can be used in human clinical trials. PMID- 17112729 TI - Inhibition of horseradish peroxidase catalytic activity by new 3-phenylcoumarin derivatives: synthesis and structure-activity relationships. AB - Twenty hydroxylated and acetoxylated 3-phenylcoumarins were synthesized, and the structure-activity relationships were investigated by evaluating the ability of these compounds to modulate horseradish peroxidase (HRP) catalytic activity and comparing the results to four flavonoids (quercetin, myricetin, kaempferol and galangin), previously reported as HRP inhibitors. It was observed that 3 phenylcoumarins bearing a catechol group were as active as quercetin and myricetin, which also show this substituent in the B-ring. The presence of 6,2' dihydroxy group or 6,7,3',4'-tetraacetoxy group in the 3-phenylcoumarin structure also contributed to a significant inhibitory effect on the HRP activity. The catechol-containing 3-phenylcoumarin derivatives also showed free radical scavenger activity. Molecular modeling studies by docking suggested that interactions between the heme group in the HRP active site and the catechol group linked to the flavonoid B-ring or to the 3-phenyl coumarin ring are important to inhibit enzyme catalytic activity. PMID- 17112730 TI - Effects of antioxidants on auditory nerve function and survival in deafened guinea pigs. AB - Based on in vitro studies, it is hypothesized that neurotrophic factor deprivation following deafferentation elicits an oxidative state change in the deafferented neuron and the formation of free radicals that then signal cell death pathways. This pathway to cell death was tested in vivo by assessing the efficacy of antioxidants (AOs) to prevent degeneration of deafferented CNVIII spiral ganglion cells (SGCs) in deafened guinea pigs. Following destruction of sensory cells, guinea pigs were treated immediately with Trolox (a water soluble vitamin E analogue)+ascorbic acid (vitamin C) administered either locally, directly in the inner ear, or systemically. Electrical auditory brainstem response (EABR) thresholds were recorded to assess nerve function and showed a large increase following deafness. In treated animals EABR thresholds decreased and surviving SGCs were increased significantly compared to untreated animals. These results indicate that a change in oxidative state following deafferentation plays a role in nerve cell death and antioxidant therapy may rescue SGCs from deafferentation-induced degeneration. PMID- 17112731 TI - Changes in glial glutamate transporters in human epileptogenic hippocampus: inadequate explanation for high extracellular glutamate during seizures. AB - Temporal lobe epilepsy (TLE) with hippocampal sclerosis is associated with high extracellular glutamate levels, which could trigger seizures. Down-regulation of glial glutamate transporters GLAST (EAAT1) and GLT-1 (EAAT2) in sclerotic hippocampi may account for such increases. Their distribution was compared immunohistochemically in non-sclerotic and sclerotic hippocampi and localized only in astrocytes, with weaker immunoreactivity for both transporters in areas associated with pronounced neuronal loss, especially in CA1, but no decrease or even an increase in areas with less neuronal loss, like CA2 and the subiculum in the sclerotic group. Such compensatory changes in immunoreactivity may account for the lack of differences between the groups in immunoblot studies as blots show the average concentrations in the samples. These data suggest that differences in glial glutamate transporter distribution between the two groups of hippocampi may be an insufficient explanation for the high levels of extracellular glutamate in sclerotic seizure foci observed through in vivo dialysis studies. PMID- 17112732 TI - Absence of perforin expression confers axonal protection despite demyelination. AB - Current evidence suggests that demyelination may be a necessary but not a sufficient condition for neurologic deficits associated with multiple sclerosis. Axon injury that occurs within the permissive environment of the demyelinated lesion is better correlated with functional deficits, but the mechanisms and cellular effectors of this injury are largely unknown. In an effort to identify potential axon injury mediators, we examined demyelination, motor function, and the number of spinal axons in perforin-deficient mice. Perforin is a critical molecular mediator of cytotoxic immunological injury and we hypothesized that genetic deletion of perforin expression would protect demyelinated axons. Indeed, we found that while perforin-deficient mice had considerable spinal cord demyelination 180 days after infection with Theiler's murine encephalomyelitis virus, such mice exhibited functional and axonal preservation comparable to non demyelinated perforin-competent controls. We conclude that perforin-dependent effector cells such as cytotoxic T cells, gammadelta T cells, and natural killer cells may play a role in axon damage that is dependent upon but separable from demyelination. PMID- 17112733 TI - Collateral blood flow between left coronary artery bypass grafts and chronically occluded right coronary circulation in patients with triple vessel disease. Observations during complete revascularisation of beating hearts. AB - OBJECTIVE: Preoperative measurements of collateral blood flow in patients with triple vessel disease and chronic occlusions of the right coronary artery do not, currently, ascertain the need to revascularise an occluded right coronary artery. We performed direct measurements of flow across left coronary bypass grafts to determine their contributions to collateral blood flow. METHODS: Collateral blood flow was scored preoperatively according to Rentrop in 13 patients with triple vessel disease and chronic occlusions of the right coronary artery who underwent complete, off-pump, surgical revascularisation. The transit-time flow through the left coronary grafts was measured before and after unclamping of the right coronary artery bypass graft. RESULTS: Unclamping of the right coronary artery bypass graft was associated with a 5.9+/-6.9ml/min (mean+/-SD) decrease in flow across the left circumflex territory (P=0.009), which was proportional to the preoperative Rentrop score (P=0.007). No significant change was observed in flow across the graft to the left anterior descending artery. CONCLUSIONS: Grafts to the left circumflex system are the only grafts that supply a significant, albeit modest amount of collateral blood flow to chronically occluded right coronary artery. These observations confirm that (1) most collateral flow after revascularisation is supplied by the native network, and (2) revascularisation of an occluded right coronary artery is fully justified. PMID- 17112734 TI - Proximal reconstruction during Type A aortic dissection. PMID- 17112735 TI - Tunable transition metal-ligand complexation for enhanced elucidation of flavonoid diglycosides by electrospray ionization mass spectrometry. AB - A tunable ESI-MS/MS strategy for differentiation of flavone and flavanone diglycoside isomers based on metal complexation with auxiliary ligands is reported. The addition of a metal salt and an auxiliary ligand to a flavonoid solution results in the formation of [M(II) (flavonoid-H) auxiliary ligand](+) complexes, where M(II) is a transition metal. A series of auxiliary ligands with electron-withdrawing substituents were synthesized to tailor the relative metal binding affinities of the ligands and thus directly influence the stabilities, and consequently the dissociation pathways, of the complexes. Upon collisionally activated dissociation, the complexes yield fragmentation patterns in which the abundances of key diagnostic ions are enhanced, thus facilitating isomer differentiation. PMID- 17112736 TI - Shrinking droplets in electrospray ionization and their influence on chemical equilibria. AB - We investigated how chemical equilibria are affected by the electrospray process, using simultaneous in situ measurements by laser-induced fluorescence (LIF) and phase Doppler anemometry (PDA). The motivation for this study was the increasing number of publications in which electrospray ionization mass spectrometry is used for binding constant determination. The PDA was used to monitor droplet size and velocity, whereas LIF was used to monitor fluorescent analytes within the electrospray droplets. Using acetonitrile as solvent, we found an average initial droplet diameter of 10 microm in the electrospray. The PDA allowed us to follow the evolution of these droplets down to a size of 1 microm. Rhodamine B sulfonylchloride was used as a fluorescent analyte within the electrospray. By spatially resolved LIF it was possible to probe the dimerization equilibrium of this dye. Measurements at different spray positions showed no influence of the decreasing droplet size on the monomer-dimer equilibrium. However, with the fluorescent dye pair DCM and oxazine 1 it was shown that a concentration increase does occur within electrosprayed droplets, using fluorescence resonance energy transfer as a probe for the average pair distance. PMID- 17112737 TI - Electron capture in spin-trap capped peptides. An experimental example of ergodic dissociation in peptide cation-radicals. AB - Electron capture dissociation was studied with tetradecapeptides and pentadecapeptides that were capped at N-termini with a 2-(4'-carboxypyrid-2'-yl) 4-carboxamide group (pepy), e.g., pepy-AEQLLQEEQLLQEL-NH(2), pepy-AQEFGEQGQKALKQL NH(2), and pepy-AQEGSEQAQKFFKQL-NH(2). Doubly and triply protonated peptide cations underwent efficient electron capture in the ion-cyclotron resonance cell to yield charge-reduced species. However, the electron capture was not accompanied by backbone dissociations. When the peptide ions were preheated by absorption of infrared photons close to the dissociation threshold, subsequent electron capture triggered ion dissociations near the remote C-terminus forming mainly (b(11-14) + 1)(+)* fragment ions that were analogous to those produced by infrared multiphoton dissociation alone. Ab initio calculations indicated that the N-1 and N-1' positions in the pepy moiety had topical gas-phase basicities (GB = 923 kJ mol(-1)) that were greater than those of backbone amide groups. Hence, pepy was a likely protonation site in the doubly and triply charged ions. Electron capture in the protonated pepy moiety produced the ground electronic state of the charge-reduced cation-radical with a topical recombination energy, RE = 5.43-5.46 eV, which was greater than that of protonated peptide residues. The hydrogen atom in the charge-reduced pepy moiety was bound by >160 kJ mol(-1), which exceeded the hydrogen atom affinity of the backbone amide groups (21-41 kJ mol(-1)). Thus, the pepy moiety functioned as a stable electron and hydrogen atom trap that did not trigger radical-type dissociations in the peptide backbone that are typical of ECD. Instead, the internal energy gained by electron capture was redistributed over the peptide moiety, and when combined with additional IR excitation, induced proton-driven ion dissociations which occurred at sites that were remote from the site of electron capture. This example of a spin-remote fragmentation provided the first clear-cut experimental example of an ergodic dissociation upon ECD. PMID- 17112738 TI - Accurate mass determination of organotrifluoroborates. AB - Exact mass measurements were obtained for a variety of potassium- and tetra-n butylammonium organotrifluoroborates using commercially available organic sulfate salts as internal reference standards. Accuracies were determined within 5 ppm using a sector ESI mass spectrometer operating in the negative ionization mode. PMID- 17112739 TI - Motexafin gadolinium: a novel redox active drug for cancer therapy. AB - Motexafin gadolinium (MGd, Xcytrin) is an aromatic macrocycle that has a strong affinity for electrons, i.e., it is easily reduced. In the presence of oxygen, MGd accepts electrons from various cellular reducing metabolites and forms superoxide and other reactive oxygen species (ROS) by redox cycling. The reaction with NADPH is dramatically accelerated by various oxido-reductases including thioredoxin reductase. In vitro studies with various cancer cell lines have shown an increase in ROS and intracellular free zinc in cells treated with MGd. MGd increases cytotoxicity of ionizing radiation and various chemotherapy agents and may be directly cytotoxic to tumor cells under certain conditions. MGd selectively localizes in tumors, perhaps due to their metabolic perturbations. MGd treatment in murine models enhances tumor response to radiation and chemotherapy agents. In controlled, randomized clinical trials, combining MGd treatment with ionizing radiation improves time to neurologic progression in lung cancer patients with brain metastases. The molecular target for MGd appears to be thioredoxin reductase which, when inhibited, results in cellular redox stress, cytotoxicity and an increase in tumor responsiveness to a variety of treatments. PMID- 17112740 TI - Longitudinal analysis of gray and white matter loss in patients with systemic lupus erythematosus. AB - Cerebral atrophy has been described to occur in systemic lupus erythematosus (SLE) with variable frequency. The aim of this study was to determine white and gray matter abnormalities in brain magnetic resonance imaging (MRI) of patients with SLE and to determine if these abnormalities progress over a one-year period. Seventy-five patients with SLE and 44 healthy age and sex-matched controls were enrolled in this study. T1-weighted volumetric images were used for voxel based morphometry (VBM) analyses. SLE patients exhibited a significant reduction in white matter and gray matter volume compared to controls (p=0.001). Follow-up images, after an average interval of 19 months, revealed a progressive white matter and gray matter atrophy (p=0.001). Reduced white and gray matter volume was associated with disease duration and the presence of antiphospholipid antibodies. Patients with severe cognitive impairment had a more pronounced white and gray matter reduction than patients with moderate cognitive impairment. Total corticosteroid dose was associated with gray matter reduction and not with white matter loss in SLE patients. We concluded that brain tissue loss associated with SLE is significant and progresses over a relatively short period of time. Disease duration, the presence of antiphospholipid antibodies and cognitive impairment were associated with white and gray matter loss. Corticosteroid was associated only with gray matter atrophy. PMID- 17112741 TI - Investigating the functional interaction between semantic and episodic memory: convergent behavioral and electrophysiological evidence for the role of familiarity. AB - Throughout our lives we acquire general knowledge about the world (semantic memory) while also retaining memories of specific events (episodic memory). Although these two forms of memory have been dissociated on the basis of neuropsychological data, it is clear that they typically function together during normal cognition. The goal of the present study was to investigate this interaction. One influence of semantic memory on episodic retrieval is 'Levels Of Processing'; recognition is enhanced when stimuli are processed in a semantically meaningful way. Studies examining this semantic processing advantage have largely concluded that semantic memory augments episodic retrieval primarily by enhancing recollection. The present study provides strong evidence for an alternative relationship between semantic and episodic memory. We employed a manipulation of the semantic coherence of to-be-remembered information (semantically related vs. unrelated word pairs) during an associative recognition memory test. Results revealed that associative recognition is significantly enhanced for semantically coherent material, and behavioral estimates (using the process dissociation procedure) demonstrated concomitant changes in the contribution of familiarity to retrieval. Neuroimaging data (event-related potentials recorded at test) also revealed a significant increase in familiarity based retrieval. The electrophysiological correlate of familiarity (the mid-frontal ERP old/new effect) was larger for semantically related compared to unrelated word pairs, but no difference was present in the electrophysiological correlate of recollection (the left parietal old/new effect). We conclude that semantic memory and episodic memory do indeed interact in normal functioning, and not only by modulating recollection, but also by enhancing familiarity. PMID- 17112742 TI - Motor imagery of complex everyday movements. An fMRI study. AB - The present study aimed to investigate the functional neuroanatomical correlates of motor imagery (MI) of complex everyday movements (also called everyday tasks or functional tasks). 15 participants imagined two different types of everyday movements, movements confined to the upper extremities (UE; e.g., eating a meal) and movements involving the whole body (WB; e.g., swimming), during fMRI scanning. Results showed that both movement types activated the lateral and medial premotor cortices bilaterally, the left parietal cortex, and the right basal ganglia. Direct comparison of WB and UE movements further revealed a homuncular organization in the primary sensorimotor cortices (SMC), with UE movements represented in inferior parts of the SMC and WB movements in superior and medial parts. These results demonstrate that MI of everyday movements drives a cortical network comparable to the one described for more simple movements such as finger opposition. The findings further are in accordance with the suggestion that motor imagery-based mental practice is effective because it activates a comparable cortical network as overt training. Since most people are familiar with everyday movements and therefore a practice of the movement prior to scanning is not necessarily required, the current paradigm seems particularly appealing for clinical research and application focusing on patients with low or no residual motor abilities. PMID- 17112743 TI - Focal cortical atrophy in multiple sclerosis: relation to lesion load and disability. AB - Multiple sclerosis (MS) is thought to predominantly affect white matter (WM). Recently, however, loss of cortical gray matter has also been described. Little is known about the cause of cortical atrophy in MS, whether it occurs early in the disease course, and whether it affects all cortical regions equally or if there is a preferential pattern of focal cortical atrophy. An automated method was used to compute the thickness at every vertex of the cortical surface of the brains of 425 early relapsing-remitting MS patients. We correlated cortical thickness with the WM lesion load and the Expanded Disability Status Scale score. Mean cortical thickness correlated with WM lesion load and disability. The correlations of cortical thickness with total lesion load and disability were most significant in cingulate gyrus, insula, and associative cortical regions. Conversely, primary sensory, visual, and motor areas showed a less significant relationship. The highest amount of atrophy per lesion volume or disability scale unit was in the anterior cingulate cortex. This study confirms the relation between cortical atrophy, WM lesion load, and disability in MS, and suggests that cortical atrophy occurs even in MS patients with only mild disability. Most interestingly, we show a specific regional pattern of focal atrophy in MS that is distinctively different from the one in normal aging. The predilection of the atrophic process for areas that are heavily inter-connected with other brain regions suggests that interruption of WM tracts by MS plaques contributes, at least in part, to the development of cortical atrophy. PMID- 17112744 TI - Functional local connections with differential activity-dependence and critical periods surrounding the primary auditory cortex in rat cerebral slices. AB - Sensory information is processed in neural networks connecting the primary sensory cortices with surrounding higher areas. Here, we investigated the properties of local connections between the primary auditory cortex (area 41) and surrounding areas (areas 20, 36, 18a and 39) in rat cerebral slices. Neural activities elicited by repetitive electrical stimulation were visualized using the activity-dependent changes in endogenous fluorescence derived from mitochondrial flavoproteins, which mostly reflect activities produced by polysynaptic glutamatergic transmission. Polysynaptic feedforward propagation was dominant compared with the corresponding polysynaptic feedback propagation between the primary (area 41) and secondary (areas 20 and 36) auditory cortices, while such a tendency was less clear in other pathways. Long inter-areal (>1 mm) propagation with the same dominancy was observed after layer V stimulation between areas 41 and 20, and was not affected by cutting the underlying white matter. Activity-dependent changes in neural activities induced by low-frequency stimulation in the presence of 1 microM bicuculline were investigated using Ca2+ imaging. Significant potentiation of the polysynaptic Ca2+ activities was only observed in polysynaptic feedforward pathways from the primary to secondary auditory cortices. Experience-dependence of the connections between areas 41 and 20 was investigated using flavoprotein fluorescence imaging. The activities from areas 41 to 20 were reduced by cochlear lesions produced at P12 but not at P28, while the activities from areas 20 to 41 were reduced by the lesions at P28, suggesting the critical period for the polysynaptic feedforward connection was before P28, while for the polysynaptic feedback connection was after P28. PMID- 17112745 TI - Performance on an episodic encoding task yields further insight into functional brain development. AB - To further characterize changes in functional brain development that are associated with the emergence of cognitive control, participants 14 to 28 years of age were scanned while performing an episodic encoding task with a levels-of processing manipulation. Using data from the 12 youngest and oldest participants (endpoint groups), 18 regions were identified that showed group differences in task-related activity as a function of processing depth. One region, located in left inferior frontal gyrus, showed enhanced activity in deep relative to shallow encoding that was larger in magnitude for the older group. Seventeen regions showed enhanced activity in shallow relative to deep encoding that was larger in magnitude for the youngest group. These regions were distributed across a broad network that included both cortical and subcortical areas. Regression analyses using the entire sample showed that age made a significant contribution to the difference in beta weights between deep and shallow encoding for 17 of the 18 identified regions in the direction predicted by the endpoint analysis. We conclude that the patterns of brain activation associated with deep and shallow encoding differ between adolescents and young adults in a manner that is consistent with the interactive specialization account of functional brain development. PMID- 17112746 TI - Improved quality of auditory event-related potentials recorded simultaneously with 3-T fMRI: removal of the ballistocardiogram artefact. AB - EEG signals recorded simultaneously with fMRI are massively compromised by severe artefacts, among them the cardiac cycle-related ballistocardiogram (BCG) artefact. Different methods have been proposed to remove the BCG artefact focusing on channel-wise template subtraction procedures or spatial filtering approaches such as independent component analysis (ICA). Here we systematically compared the performance of the optimal basis set (OBS), a channel-wise correction approach, with ICA and a recently proposed combination of both (OBS ICA). The three different procedures were applied to 60-channel EEG data from 12 subjects recorded during fMRI acquisition in a 3-T scanner. In addition to examination of the residual BCG artefact, the signal-to-noise ratio (SNR) and the topography of the resulting auditory evoked potential component N1 were compared. Whereas all three approaches led to a significant artefact reduction, the ICA procedure resulted in a significantly reduced N1 SNR and amplitude when compared to BCG-uncorrected data, indicating a rather poor performance. In contrast to ICA, OBS and OBS-ICA corrected data substantially improved the SNR of the N1. The quality of the auditory evoked potential N1 topography was investigated by means of equivalent current dipole modelling. On a descriptive level, all three correction procedures led to a reduced localization error when compared to BCG uncorrected data. This improvement was significant for OBS-ICA. We conclude that OBS and OBS-ICA can efficiently remove BCG artefacts and substantially improve the quality of EEG signals recorded inside the scanner, a prerequisite for the successful integration of simultaneously recorded EEG and fMRI. PMID- 17112747 TI - Complete artifact removal for EEG recorded during continuous fMRI using independent component analysis. AB - The simultaneous recording of EEG and fMRI is a promising method for combining the electrophysiological and hemodynamic information on cerebral dynamics. However, EEG recordings performed in the MRI scanner are contaminated by imaging, ballistocardiographic (BCG) and ocular artifacts. A number of processing techniques for the cancellation of fMRI environment disturbances exist: the most popular is averaged artifact subtraction (AAS), which performs well for the imaging artifact, but has some limitations in removing the BCG artifact, due to the variability in cardiac wave duration and shape; furthermore, no processing method to attenuate ocular artifact is currently used in EEG/fMRI, and contaminated epochs are simply rejected before signal analysis. In this work, we present a comprehensive method based on independent component analysis (ICA) for simultaneously removing BCG and ocular artifacts from the EEG recordings, as well as residual MRI contamination left by AAS. The ICA method has been tested on event-related potentials (ERPs) obtained from a visual oddball paradigm: it is very effective in attenuating artifacts in order to reconstruct clear brain signals from EEG acquired in the MRI scanner. It performs significantly better than the AAS method in removing the BCG artifact. Furthermore, since ocular artifacts can be completely suppressed, a larger number of trials is available for analysis. A comparison of ERPs inside the magnetic environment with those obtained out of the MRI scanner confirms that no systematic bias in the ERP waveform is produced by the ICA method. PMID- 17112748 TI - Ictal source analysis: localization and imaging of causal interactions in humans. AB - We propose a new integrative approach to characterize the structure of seizures in the space, time, and frequency domains. Such characterization leads to a new technical development of ictal source analysis for the presurgical evaluation of epilepsy patients. The present new ictal source analysis method consists of three parts. First, a three-dimensional source scanning procedure is performed by a spatio-temporal FINE source localization method to locate the multiple sources responsible for the time evolving ictal rhythms at their onsets. Next, the dynamic behavior of the sources is modeled by a multivariate autoregressive process (MVAR). Lastly, the causal interaction patterns among the sources as a function of frequency are estimated from the MVAR modeling of the source temporal dynamics. The causal interaction patterns indicate the dynamic communications between sources, which are useful in distinguishing the primary sources responsible for the ictal onset from the secondary sources caused by the ictal propagation. The present ictal analysis strategy has been applied to a number of seizures from five epilepsy patients, and their results are consistent with observations from either MRI lesions or SPECT scans, which indicate its effectiveness. Each step of the ictal source analysis is statistically evaluated in order to guarantee the confidence in the results. PMID- 17112749 TI - Automated template-based PET region of interest analyses in the aging brain. AB - The definition of regions of interest for PET data analysis poses a number of complex problems. While studies have shown that regions drawn on a template can be appropriate for extracting data for normal healthy subjects, it is unclear how these results can be applied to different populations. In this study, we focused on the aging population and examined how different parameters in the template data-extraction process may affect the accuracy of the results. We first present an automated method for extracting PET counts using a region-of-interest approach within a template framework. Then, we discuss two studies in which we measure the effects of varying specific parameters in this process. In study 1 we examined three parameters that may influence this process: choice of template, region, and threshold. In study 2 we focused on the hippocampus. We considered 6 different templates, and examined how well the subject-specific hippocampal masks overlapped with each other and with the template hippocampal masks after normalization. While the data in the older cohort are more variable than the normal population, the results suggest that using an appropriate template and selecting the correct parameters for the template-based ROI method can provide template-extracted counts that are highly correlated to counts extracted using subject-specific ROIs. PMID- 17112750 TI - Neural correlates of a 'pessimistic' attitude when anticipating events of unknown emotional valence. AB - Since we do not know what future holds for us, we prepare for expected emotional events in order to deal with a pleasant or threatening environment. From an evolutionary perspective, it makes sense to be particularly prepared for the worst-case scenario. We were interested to evaluate whether this assumption is reflected in the central nervous information processing associated with expecting visual stimuli of unknown emotional valence. While being scanned with functional magnetic resonance imaging, healthy subjects were cued to expect and then perceive visual stimuli with a known emotional valence as pleasant, unpleasant, and neutral, as well as stimuli of unknown valence that could have been either pleasant or unpleasant. While anticipating pictures of unknown valence, the activity of emotion processing brain areas was similar to activity associated with expecting unpleasant pictures, but there were no areas in which the activity was similar to the activity when expecting pleasant pictures. The activity of the revealed regions, including bilateral insula, right inferior frontal gyrus, medial thalamus, and red nucleus, further correlated with the individual ratings of mood: the worse the mood, the higher the activity. These areas are supposedly involved in a network for internal adaptation and preparation processes in order to act according to potential or certain unpleasant events. Their activity appears to reflect a 'pessimistic' bias by anticipating the events of unknown valence to be unpleasant. PMID- 17112752 TI - Co-sleeping, an ancient practice: issues of the past and present, and possibilities for the future. AB - Co-sleeping-infants sharing the mother's sleep space-has prevailed throughout human evolution, and continued over the centuries of western civilization despite controversy and blame of co-sleeping mothers for the deaths of their infants. By the past century, "crib death" was recognized, later identified as Sudden Infant Death Syndrome (SIDS), and generally found to occur more frequently during bed sharing. Pediatricians warned parents of the dangers of SIDS and other risks of bed sharing, and the frequency of bed sharing decreased markedly over the years. However, during recent decades, bed sharing began to increase, though major issues were raised, including: whether bed sharing actually exacerbates or is protective against the occurrence of SIDS, whether the practice facilitates breast feeding, whether bed sharing is beneficial for an infant's development, and other concerns. Dissention may soon be diminished by use of a crib which opens at the mother's bed-side and is becoming a popular approach to mother-and infant closeness through the night. PMID- 17112751 TI - Stromal cell networks regulate lymphocyte entry, migration, and territoriality in lymph nodes. AB - After entry into lymph nodes (LNs), B cells migrate to follicles, whereas T cells remain in the paracortex, with each lymphocyte type showing apparently random migration within these distinct areas. Other than chemokines, the factors contributing to this spatial segregation and to the observed patterns of lymphocyte movement are poorly characterized. By combining confocal, electron, and intravital microscopy, we showed that the fibroblastic reticular cell network regulated naive T cell access to the paracortex and also supported and defined the limits of T cell movement within this domain, whereas a distinct follicular dendritic cell network similarly served as the substratum for movement of follicular B cells. These results highlight the central role of stromal microanatomy in orchestrating cell migration within the LN. PMID- 17112754 TI - Multiple echo NMR velocimetry: fast and localized measurements of steady and pulsatile flows in small channels. AB - The understanding of fluid transport in miniaturized flow devices is an important component in the design of flow cells, micromixers, and microreactors. In this manuscript, we employ NMR in the form of a voxel-selective multiple modulation multiple echo sequence (MMMEV) to monitor average velocities in individual microchannels inside a six-channel network. The technique produces average velocities which are consistent with the imposed flow rates. In addition, we take advantage of the short acquisition time (32 ms per velocity component) of the technique to quantitatively track the time evolution of the fluid velocity in a pulsatile flow phantom. PMID- 17112755 TI - Perkinsus marinus, a protozoan parasite of the eastern oyster, has a requirement for dietary sterols. AB - Perkinsus marinus, a protozoan parasite of the eastern oyster, Crassostrea virginica, causes high mortality in its host along the Atlantic and Gulf coasts of North America. P. marinus meronts cultured in vitro in medium containing complete lipid supplement (cod liver oil, cholesterol and alpha tocopherol acetate in detergent) are able to synthesize a wide variety of lipids, yet cultures cannot be maintained in lipid-free medium. To determine P. marinus lipid requirements meronts were inoculated into media containing different combinations of lipid components in detergent. Treatments included complete lipid supplement (positive control), detergent only (negative control), cholesterol in detergent, alpha tocopherol acetate in detergent and cholesterol+alpha tocopherol acetate in detergent. Meronts proliferated in the positive control medium and media containing cholesterol or cholesterol+alpha tocopherol acetate, but failed to proliferate in the negative control medium and the medium containing just alpha tocopherol acetate. Gas chromatography analysis of P. marinus meronts grown in medium with added (13)C sodium acetate (0.5 mg mL(-1)) revealed the presence of fatty acids containing (13)C, but the only sterol present was cholesterol containing no (13)C. These results suggest that P. marinus cannot synthesize sterols and must sequester them from its host. PMID- 17112756 TI - Digestive enzymes and metabolic profile of Labeo rohita fingerlings fed diets with different crude protein levels. AB - Labeo rohita, commonly called rohu is one of the most important fish species for aquaculture in India. Digestive enzyme response and metabolic profile of fingerling L. rohita to different dietary crude protein (CP) levels (viz. 25, 30, 35 and 40%) were studied in an attempt to optimize a practical diet formulation for this species. After 45 days of feeding, activity of digestive enzymes and metabolite concentrations were assayed. Amylase, lipase and alkaline phosphatase (ALP) activities were not influenced by the dietary protein, but proteolytic and acid phosphatase (ACP) activities varied (P<0.05) between the treatments. Proteolytic activity showed a second order polynomial relationship with dietary crude protein (CP) as Y = 0.0734X(2) + 4.937X - 68.37, r(2)=0.97. A positive correlation was observed between dietary CP and amylase (r(2)=0.78). All the metabolites except muscle glucose showed significant change corresponding to the dietary protein levels. Glucose and glycogen levels corresponded to the dietary carbohydrate levels. Muscle and plasma pyruvic acid increased as the crude protein in the diet increased, whereas liver pyruvic acid showed the opposite trend. Muscle protein content was not affected by dietary CP. Protein fractions in plasma (total protein, albumin and globulin) showed maximum values in 30% CP fed group. It is concluded that proteolytic activity and ACP are the major digestive enzymes responsive to dietary CP in L. rohita fingerlings. Considering the cost effectiveness of the diet, and based on liver and plasma free amino acid levels and plasma protein fractions, 30% crude protein is recommended as the optimal dietary protein for L. rohita fingerlings. PMID- 17112757 TI - Hydrolysis and fermentation of amorphous cellulose by recombinant Saccharomyces cerevisiae. AB - In this study, we expressed two cellulase encoding genes, an endoglucanase of Trichoderma reesei (EGI) and the beta-glucosidase of Saccharomycopsis fibuligera (BGL1), in combination in Saccharomyces cerevisiae. The resulting strain was able to grow on phosphoric acid swollen cellulose (PASC) through simultaneous production of sufficient extracellular endoglucanase and beta-glucosidase activity. Anaerobic growth (0.03h(-1)) up to 0.27gl(-1) DCW was observed on medium containing 10gl(-1) PASC as sole carbohydrate source with concomitant ethanol production of up to 1.0gl(-1). We have thus demonstrated the construction of a yeast strain capable of growth on and one-step conversion of amorphous cellulose to ethanol, representing significant progress towards realization of one-step processing of cellulosic biomass in a consolidated bioprocessing configuration. To our knowledge, this is the first report of a recombinant strain of S. cerevisiae growing on pure cellulose. PMID- 17112758 TI - [Staging methods]. AB - There have been main changes in the methods for the work-up of lung cancer over the last 15 years. Guidelines of the main scientific societies are reviewed. The new methods are discussed: brain nuclear magnetic resonance (NMR), positron emission tomography (PET scan), mediastinal nodes biopsy by echoguided endoscopy. These new techniques will change the classical approach as recommended by ATS/ERS. PMID- 17112759 TI - Relationship between 1,25-dihydroxycholecalciferol levels and functional outcome after hip fracture in elderly patients. AB - A prospective study was performed in 109 patients with osteoporotic hip fracture to verify the relationship between 1,25-dihydroxycholecalciferol levels at admission and functional recovery in elderly patients with hip fracture. 47.7% of all the patients had 1,25-dihydroxycholecalciferol levels below the reference values of our laboratory. At 1 year of the fracture only 33 patients (30.3%) had recovered the functional level they had before the fracture. In the bivariate analysis, a statistically significant relationship was found between reduced 1,25 dihydroxycholecalciferol levels and a poorer functional recovery 1 year after the fracture was sustained. A multivariate analysis showed a significant relationship between 1,25-dihydroxycholecalciferol levels and absolute dependence of the patient 1 year after the fracture (P = 0.005; OR 6.97: CI- 1.77-27.41). PMID- 17112761 TI - Severity of injuries among sexual assault victims. AB - It is generally accepted that victims of sexual assault sustain bodily injury. This study's objective was to determine specific characteristics and severity of injuries among victims of sexual violence in Belgrade. Retrospectively, we analyzed a subgroup of victims of sexual violence that was legally processed over a five-year period. We evaluated 113 cases of sexual crimes selected from the District Court of Belgrade in order to analyze the medical records. All victims were female, at average 24.1 years old (range 5-80 years). In more than half of the cases (52%) evaluated, a medical examination was completed on the day of assault, while 84% took place within 72 hours post-assault. Due to delayed referral, body examination was not conducted on 12 victims (10.6%). We noted one or more extra-genital injuries in 64 victims (63.4%), no injuries in 36 victims (35.6%), whereas for one victim the medical records were inconclusive. Injuries, predominantly bruises, were located on limbs (32%), face (23%), and torso (7%). Abrasions and contusions were less frequently present, while two victims sustained lacerations. The Clinical Injury Extent Score was used to rate the physical severity of the assault. The majority of victims (44%) sustained light injuries, 18% were moderate, while one victim had severe injuries. PMID- 17112762 TI - The tendency of suicide among the elderly in Mie Prefecture, Japan. PMID- 17112763 TI - The risk factors of suicide by poisoning among psychiatry department outpatients. PMID- 17112764 TI - Survival following accidental ligature strangulation: a case report. AB - Survival following accidental ligature strangulation is quite rare. The present case involves an adult male strangulated by a soft cotton cloth entangled in the rotor of a machine. Unilateral neck compression allowed survival of the victim. The victim escaped with minimal injuries which were limited to contusion of the neck and edema of the vocal cords and inter arytenoid region. PMID- 17112765 TI - Postmortem sole incisions - a new sign of heroin overdose? AB - Postmortem sole incisions have been observed in a number of heroin overdose fatalities. Acqueintance of those victims confessed to producing those incisions as a life saving procedure in a futile attempt to help the comatose overdose victim. They thought that bleeding the unconscious victim would remove the overdose, in manner similar to bloodletting or phlebotomy which is still popular in the Gulf region. The presence of such wounds has become a first indication or rather "sign" of heroin poisoning. In such cases, laboratory investigation confirmed the pathologist's preliminary suspicion. In Dubai, postmortem sole incisions are important sign of death from heroin overdose even in the absence of other classical signs. This sign becomes more credible when accompanied by other signs and/or circumstantial evidence suggestive of heroin use. It is suggested that this should be called "bloodletting sign" of death from heroin overdose. The sign should not be confused with the self-inflicted cuts seen on the arms and forearms of drug misusers which are caused for other reasons. PMID- 17112766 TI - Rape trauma syndrome: time to open the floodgates? AB - The conviction rate in rape cases remains remarkably low, leading many to believe a new approach is needed to prosecuting such crimes. Many important steps have been taken in relation to the collection and retention of evidence and the setting up of specialist rape prosecutors, but important evidential weaknesses remain present in the system alongside Victorian attitudes to sexual conduct. The British Home Office is currently consulting on a number of measures, including the admissibility of evidence showing the presence of rape trauma syndrome. This paper examines the history of this controversial evidential debate. PMID- 17112767 TI - Suicides in Northern India: comparison of trends and review of literature. AB - Trends of suicide vary widely according to time, region, age group, sex, and race. Despite mixed trends of increases or decreases in suicide rates around the world, suicide remains an important public-health problem. In an effort to understand and prevent suicide, researchers have investigated medical, psychosocial, cultural, and socio-economic risk factors associated with the environment as a promising line of research. There is now considerable evidence that childhood and family adversities in general such as childhood sexual and physical abuse, witnessing domestic violence, parental separation or divorce and living with substance abusing, mentally ill or criminal family members may be both strongly interrelated and individually related to suicidal behavior in adolescents as well as adults. The approach towards prevention of suicide has to be multidisciplinary. To recognize that adverse childhood experiences that frequently take place as multiple events, identifying and treating those young people who have been exposed to such experiences, promoting increased awareness among parents, teachers, and health professionals of the important role that severe interpersonal difficulties and dysfunctional cognitions can play in the development of suicidal behavior in young people, and helping parents modify their maladaptive child-rearing behavior could help. Child and family support programs, employment support for mothers, and legal guarantees of gender equality, could moderate problems of socio-economic disparity and poverty, which predicts both parents' and children's suicidal behaviors in modern societies. PMID- 17112768 TI - The diagnostic validity of the cervical flexion-rotation test in C1/2-related cervicogenic headache. AB - This single-blind comparative group design aimed to investigate the sensitivity and specificity of the cervical flexion-rotation test in the diagnosis of C1/2 related cervicogenic headache. This study tested 23 cervicogenic headache, 23 asymptomatic controls and 12 migraine with aura subjects, all aged 18-66 years. In stage 1, an experienced manipulative physiotherapist who did not partake in the flexion-rotation test procedure identified C1/2 dysfunction using passive segmental mobility tests in the cervicogenic headache group. Those with C1/2 dysfunction participated in stage 2. In stage 2, using the flexion-rotation test, subjects were tested by two experienced manipulative physiotherapists blinded to the subjects' group allocation. Each therapist stated whether the test was positive or not based on the therapist's interpretation of range of motion. The sensitivity and specificity of the flexion-rotation test was 91% and 90%, respectively (P<.001), with an overall diagnostic accuracy of 91% (P<.001). The cervical flexion-rotation test significantly assists in the differential diagnosis of cervicogenic headache and in the identification of movement impairment at the C1/2 segment in patients with cervicogenic headache. PMID- 17112769 TI - Evaluation of optimal drug concentration in histoculture drug response assay in association with clinical efficacy for head and neck cancer. AB - Induction chemotherapy or concomitant chemoradiotherapy has been used increasingly to improve survival, organ preservation and function in patients with head and neck cancer (HNC). However, this regimen encounters significant side effects with potential adverse reactions causing many disadvantages for patients. Therefore, reliable chemosensitivity assays are needed to accurately predict the response to chemotherapy and guide the selection and treatment of patients with HNC. The main purpose of this study was to examine the optimal drug concentrations for evaluating in vitro chemosensitivity using the histoculture drug response assay (HDRA). The tested tumor specimens included 7 from oral cavities (14.3%), 12 from oropharynx (24.5%), 10 hypopharynx (20.4%), 3 larynx (6.1%), 5 sinonasal (10.2%), 2 salivary glands (4.1%), and 10 from metastatic lymph nodes (20.4%), respectively. Histopathologic types of all 49 specimens were squamous cell carcinoma. We investigated the optimal drug concentrations in HDRA searching at doses of 4-100 microg/ml for cisplatin and 60-1500 microg/ml for 5 FU. We considered the concentration of 20 microg/ml to be appropriate for evaluating cisplatin sensitivity in HNC among the tested dosages. As for cisplatin sensitivity in vitro, the 50% cut-off inhibition index (I.I.) was found to have a significant association with the clinical response to chemotherapy, with an accurate prediction rate of 77.8%. The HDRA shows a predictive value for chemosensitivity in HNC patients using the optimal drug concentration cut-off with this site specificity. PMID- 17112770 TI - The possible premalignant character of oral lichen planus and oral lichenoid lesions: a prospective five-year follow-up study of 192 patients. AB - Recently, we reported the preliminary results of a prospective study on the possible premalignant character of oral lichen planus (OLP) and oral lichenoid lesions (OLL). Based on these data it was concluded that there was some but no convincing support for the hypothesis that patients with OLL have an increased risk of development of oral cancer, but not so in patients with OLP. In the present treatise the results of prolonged follow-up of this cohort of patients have been described. A study group of 192 patients, 67 patients diagnosed with OLP and 125 patients with OLL, according to revised World Health Organization diagnostic criteria, was followed for periods ranging from 7.6 to 96.9 months (mean, 55.9 months). The expected number of patients with oral cancer in the group of patients with OLP and in the group of patients with OLL was estimated by comparing the number of patients, their ages, sex, and the length of follow-up to annual incidence rates of oral cancer for the general population in The Netherlands. The binomial test was used to determine whether the observed number of cases of cancer in the OLP group and the OLL group exceeded the expected numbers. Four out of 192 patients, two men and two women, developed a squamous cell carcinoma of the oral mucosa during follow-up. All malignant transformations occurred in the OLL group. The malignant transformation of the OLL group, based on a mean follow-up of 53.8 months, was calculated at 0.71% per year. A comparison of the expected against actual figures for the development of carcinomas revealed no increase in patients with OLP and a 142-fold increase in patients with OLL, the latter being statistically significant, with a p-value of 0.044. The present data give support to the hypothesis that patients with OLL have an increased risk of development of oral cancer. There seems to be no increased risk in patients with OLP. In view of our results we advise to monitor only the subgroup of OLL patients twice a year for early detection of possible malignant transformation. PMID- 17112771 TI - Evolution of the management of laryngeal cancer. AB - The treatment of laryngeal cancer has evolved through several phases, starting with wide extirpative surgical resection, and evolving through an era of conservation surgery and, finally, planned treatment using modalities of irradiation, chemotherapy and surgery in various combinations. Attempts to extirpate laryngeal cancer date to the nineteenth century, but only by the mid twentieth century did advances in anesthesia, blood transfusion and antibiotics, make this surgery safe and reliable. Techniques of partial laryngectomy by external approach developed in the second half of the twentieth century, and endoscopic use of the laser refined the concept and provided a new paradigm for surgical treatment, particularly for early lesions. During most of this era, radiation was employed as an alternative method of treatment, with surgery reserved for salvage of radiation failure. By the last decade of the twentieth century, and to the present time, the value of combined modality therapy, using planned combinations of irradiation, chemotherapy and surgery became the standard of care for advanced laryngeal cancer, permitting maximal laryngeal preservation with the highest attainable cure rates. PMID- 17112772 TI - The uncertainty of the surgical margin in the treatment of head and neck cancer. AB - We discuss our surgical philosophy concerning the subtle interplay between the size of the surgical margin taken and the resultant morbidity from ablative oncological procedures, which is ever more evident in the treatment of head and neck malignancy. The extent of tissue resection is determined by the "trade off" between cancer control and the perioperative, functional and aesthetic morbidity and mortality of the surgery. We also discuss our dilemmas concerning recent minimally invasive endoscopic microsurgical techniques for the trans-oral laser removal or co-ablation of aero-digestive tract tumours, which result in a minimal surgical margin of oncological clearance. By a process of inductive argument as to the nature of the surgical margin, we consider whether the risks of taking a lesser margin with adjuvant therapy is justified by the attendant gain in reduced surgical morbidity and the possible costs in tumour control. PMID- 17112773 TI - Clinical significance of intrathoracic lesions detected by 18F-fluorodeoxyglucose positron emission tomography in the management of patients with head and neck cancer. AB - Few studies have used positron emission tomography (PET) to identify metastases or simultaneous thoracic malignancies in patients with head and neck cancer (HNC). We retrospectively investigated the role of (18)F-fluorodeoxyglucose (FDG) positron emission tomography (PET) in detecting thoracic malignancies in patients with previously untreated HNC. Patients (n=86) with HNC and intrathoracic lesions on PET were divided into those who had abnormal FDG uptake in the mediastinum (n=29), lungs (n=34), or both (n=23). Whole body PET and chest computed tomography (CT) results were blindly reviewed and scored by two observers. The accuracy of FDG PET and CT were drawn from patients in whom diagnosis was confirmed, by histopathology or follow-up imaging, and risk factors for thoracic malignancy were analyzed. Malignancy was suspected in 23 of 86 patients (27%) with FDG uptake. Most of the lesions (83%) with abnormal FDG uptake were benign, with thoracic malignancy confirmed in 15 patients (17%). The overall sensitivity, specificity, and accuracy of FDG PET for intrathoracic malignancy in these patients were 80%, 85%, and 84%, respectively. The likelihood of thoracic malignancy in the HNC patients was associated with high FDG uptake of thoracic lesions. FDG PET may reveal lung and mediastinal malignancies with high accuracy in patients with HNC. The thoracic staging by FDG PET may be helpful in therapeutic planning for these patients. PMID- 17112774 TI - Epidermal growth factor receptor (EGFR) polymorphisms and survival in head and neck cancer patients. AB - EGFR overexpression has been implicated in the development of head and neck squamous cell carcinoma (HNSCC). This study evaluates the prognostic ability of four polymorphisms in EGFR gene for patients diagnosed with HNSCC and treated with chemoradiation. EGFR polymorphisms in the promoter region were not associated with clinical or pathological characteristics. In relation to R497K polymorphism, patients with the Arg/Arg genotype showed the highest risk of disease-specificity mortality and none of the patients with the Lys/Lys genotype died throughout the follow-up period of the study. Patients with (CA)(n) repeats <17 in both alleles tended toward inferior overall survival compared with those with (CA)(n) repeats > or = 17 in both alleles (p=0.07). Moreover, the distribution of patients with any (CA)(n) repeats > or = 17 and both alleles <17 was statistically different across patients who were recorded as having partial response or no response to therapy (p=0.034). Combination analysis of both polymorphisms, (CA)(n) repeats and R497K, suggests that these polymorphisms may be associated with clinical outcome in patients treated with chemoradiation. PMID- 17112775 TI - F-18 FDG-PET as a routine surveillance tool for the detection of recurrent head and neck squamous cell carcinoma. AB - In order to determine the efficacy and proper timing of routine PET scans for surveillance of recurrent head and neck squamous cell carcinoma (HNSCC), we evaluated the diagnostic performance of routine PET scans in relation to time interval from completion of treatment. Amongst 206 retrospectively evaluated post treatment PET scans of 159 patients with HNSCC, 156 were performed for routine surveillance in subclinical cases. Diagnostic performance of PET scan and follow up outcome were evaluated in relation to the time interval (2-6months, 6 12months, 12-24months, and >24months) of PET scan from the completion of treatment. Overall sensitivity and NPV of these PET scans for recurrence were 92.5% and 94.8%, compared with 55.0% and 76.9% for conventional evaluation methods. In the 156 routine scans, the diagnostic sensitivity, specificity, and NPV for locoregional recurrence were 90%, 91% and 97%, respectively, and the values for distant metastases and second primary cancers were 100%, 97% and 100%, respectively. The diagnostic accuracy of routine PET scans was not significantly altered by the time interval. Most (97%) of true negative cases on routine PET scans had no recurrence during a median 14months follow-up. PET scan may be a useful tool in routine surveillance for detection of recurrence in subclinical patients. For routine surveillance, the initial PET scan should be performed within 6months after completion of treatment and the proper timing of next routine PET scan for subclinical patient with initial negative PET result might be 1year after initial PET scan. PMID- 17112776 TI - The influence of clinical and demographic risk factors on the establishment of head and neck squamous cell carcinoma cell lines. AB - The purpose of this study was to generate stable cell cultures from head and neck squamous cell carcinomas (HNSCC), and retrospectively analyze the factors associated with successful cell line establishment. Fifty-two HNSCC cell lines were isolated from a series of 199 tumors collected between 1992 and 1997 at the University of Pittsburgh Medical Center. Cell lines were characterized at the molecular and cellular level to determine the features associated with cell line formation. Successful cell line formation was dependent on multiple factors, including gene amplification involving chromosomal band 11q13, local and/or regional involvement of lymph nodes, and alcohol usage. The establishment of HNSCC cell lines enriches the resources available for cancer research. Our findings indicate that generation of stable cell lines from HNSCC is biased towards tumors with a poor prognosis. Our 52 stable lines comprise one of the largest series of HNSCC cell lines in the literature, with complete demographic, histopathologic, clinical, and survival data. PMID- 17112777 TI - The influence of tamoxifen on growth behavior and cell-cell adhesion in OSCC in vitro. AB - The aim of this study was to evaluate the influence of tamoxifen on the growth and aggregation behavior, focusing on the expression pattern of E-cadherin and beta-catenin, in oral squamous cell carcinoma (OSCC) in vitro. Oral squamous cancer cell lines (UM-SCC-14A, UM-SCC-14B and UM-SCC-14C) were treated with various concentrations of tamoxifen. Growth and aggregation behavior as well as the protein expression and its changes were analysed. All cell lines are estrogen receptor (ER) positive. Tamoxifen induced a significant growth inhibition and induced the ability to form cell aggregates. This phenomena was not accompanied by a change in E-cadherin or beta-catenin expression or due to transcriptional changes. beta-catenin showed isolated membrane staining and nuclear distribution in all cell lines. A defective Ecadherin/beta-catenin complex was seen in UM-SCC 14C with no restoration through tamoxifen treatment. The cell-cell formation is increased in all cell lines without any alterations in the functional and quantitative status of E-cadherin or beta-catenin, indicating that novel cell cell adhesion complexes not involving the classical E-cadherin/beta-catenin influence cell growth and intercellular adhesion in OSCC. PMID- 17112778 TI - Increased frequency of micronuclei in peripheral blood lymphocytes of subjects infected with Helicobacter pylori. AB - Epidemiological studies have demonstrated a close association between infection with Helicobacter pylori and the development of gastric carcinoma and mucosa associated lymphoid tissue lymphomas in humans. The cytokinesis-block micronucleus assay was performed on peripheral blood lymphocytes of H. pylori infected patients in order to investigate the possible induction of genotoxic damage. The study group consisted of 70 infected subjects including 33 women and 37 men, and 66 healthy controls (37 females and 29 males). Our results indicate that in the infected group the overall frequency of binucleated micronucleated cells (BNMN) per 1000 cells was higher (17.65+/-1.55) than in the controls (7.39+/-0.66), this difference being statistically significant. No differences were found between the infected and control groups regarding the cytokinesis block proliferation index (CBPI). When the effect of different counfounding factors was evaluated, mutivariate statistical analysis revealed that age and alcohol consumption modulated the frequency of BNMN in infected people, and the interaction between alcohol use-smoking-infection also affected the BNMN frequency in H. pylori patients. Our results indicate that infection by H. pylori is associated with an increased level of cytogenetic damage in the cells of the host. PMID- 17112779 TI - The story behind the story: physician skepticism about relying on clinical information technologies to reduce medical errors. AB - PURPOSE: In order to better understand physicians' perspectives about the use of clinical information technology (CIT) to reduce medical errors, we asked physicians about opportunities and issues around clinical use of computerized physician order entry (CPOE) systems, order sets within CPOE, and handheld computers (HHCs). METHODS: We conducted 10 focus groups including 71 physicians involved in technology implementation efforts across the US between April 2002 and February 2005. RESULTS: Two major themes emerged across focus groups around reliance on CIT to reduce errors: (1) can it work? and (2) at what cost to the medical profession? Within the first theme, physicians expressed concern about the appropriateness of physician-directed CIT as a solution for medical errors, concerns regarding the current technical capabilities and level of technical support for CIT solutions, and concern about the introduction of new errors. Within the second theme, physicians were particularly concerned about time efficiency and workload redistribution associated with the introduction of CIT. Across focus groups, physicians tended to generalize about the role of all IT in their lives, potentially biasing opinions about specific technologies. CONCLUSIONS: Health care organizations attempting to promote physician use of CIT are advised to deepen consideration of physicians' perspectives about technology adoption and use in order to address their concerns, reduce skepticism, and increase the likelihood of implementation success. PMID- 17112780 TI - Characterizing 56 complete SARS-CoV S-gene sequences from Hong Kong. AB - BACKGROUND: The spike glycoprotein (S) gene of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) has been useful in analyzing the molecular epidemiology of the 2003 SARS outbreaks. OBJECTIVES: To characterize complete SARS-CoV S-gene sequences from Hong Kong. STUDY DESIGN: Fifty-six SARS CoV S-gene sequences, obtained from patients who presented with SARS to the Prince of Wales Hospital during March-May 2003, were analysed using a maximum likelihood (ML) approach, together with 138 other (both human and animal) S-gene sequences downloaded from GenBank. RESULTS: The maximum-likelihood (ML) trees showed little evolution occurring within these 56 sequences. Analysis with the other sequences, showed three distinct SARS clusters, closely correlated to previously defined early, middle and late phases of the 2003 international SARS outbreaks. In addition, two new single nucleotide variations (SNVs), T21615A and T21901A, were discovered, not previously reported elsewhere. CONCLUSIONS: The ML approach to the reconstruction of tree phylogenies is known to be superior to the more popular, less computationally and time-demanding neighbour-joining (NJ) approach. The ML analysis in this study confirms the previously reported SARS epidemiology analysed mostly using the NJ approach. The two new SNVs reported here are most likely due to the tissue-culture passaging of the clinical samples. PMID- 17112781 TI - Knowledge and attitudes about cancer pain management: a national survey of Italian oncology nurses. AB - The purpose of this study was to obtain information about the knowledge and attitudes of Italian oncology nurses concerning cancer pain management and to determine the predictors of nurses' pain management knowledge. The study was a nationwide descriptive survey and included 287 nurses in Italy from 21 oncology wards in the north, center and south of Italy. The Nurses' Knowledge and Attitudes Survey (Italian version) and a background information form were used to collect the data. Knowledge and attitudes regarding cancer pain were the main research variables. Among the 39 pain knowledge questions assessed, the mean number of correctly answered question was 21.4 (SD=5.5), with a range of 6-35. The correct answer rate for the entire scale, on average, was 55% (SD=25.9). Further analysis of items showed that more than 50% of oncology nurses underestimated the patients' pain and they did not treat it in the correct way; they also had an incorrect self-evaluation about their pain management knowledge. Results from stepwise regression showed that nurses with higher mean correct answer scores had attended more courses about pain education. There are still significant knowledge deficits and erroneous beliefs that may hamper treatment of oncology patients in pain. The results of this study could be useful to institutions involved in patient care and teaching of pain management. PMID- 17112782 TI - Implications of findings of bibliometric analyses in parasitology. PMID- 17112783 TI - Evolution of parasitic life in the ocean. PMID- 17112784 TI - ADAMTS-13 levels in fresh, stored, and solvent detergent treated plasma. AB - BACKGROUND: ADAMTS-13 is implicated in the pathophysiology of thrombotic thrombocytopenic purpura (TTP). Plasma exchange is thought to be effective through removal of a harmful substance or provision of a required material such as ADAMTS-13. As various methods are used to prepare plasma we determined the effects of storage and solvent detergent treatment on the ADAMTS-13 levels in plasma. METHODS: Samples from fresh plasma and fresh frozen plasma (FFP) were stored at 22 degrees C and ADAMTS-13 levels were measured at 0, 12, 24, 48 and 72 h. Samples were also taken from solvent detergent treated plasma (SDP) and cryosupernatant plasma (CSP). Total protein, albumin, fibrinogen and immunoglobulins were also measured. RESULTS: In fresh plasma, the levels of both the 175 and 140 Kd subunits of von Willebrand factor were consistent at 1.38 and 1.35 OD units from 0 to 48 h indicating normal ADAMTS-13 activity. The Vitex SDP produced slightly more of the 140 Kd subunit than did Octapharma SDP which gave equivalent fragments. Cryosupernatant plasma was the same as normal plasma. None of these values changed over 48 h. There was a 28% decrease in FVIII in fresh plasma over 24 h. Fibrinogen and albumin were unchanged. CONCLUSION: ADAMTS-13 levels are not significantly decreased by storage of plasma at room temperature for up to 48 h. Both CSP and SDP also contained essentially normal levels of ADAMTS-13 and therefore could be used for treatment of patients with TTP. PMID- 17112785 TI - The multi-step phosphorelay mechanism of unorthodox two-component systems in E. coli realizes ultrasensitivity to stimuli while maintaining robustness to noises. AB - E. coli has two-component systems composed of histidine kinase proteins and response regulator proteins. For a given extracellular stimulus, a histidine kinase senses the stimulus, autophosphorylates and then passes the phosphates to the cognate response regulators. The histidine kinase in an orthodox two component system has only one histidine domain where the autophosphorylation occurs, but a histidine kinase in some unusual two-component systems (unorthodox two-component systems) has two histidine domains and one aspartate domain. So, the unorthodox two-component systems have more complex phosphorelay mechanisms than orthodox two-component systems. In general, the two-component systems are required to promptly respond to external stimuli for survival of E. coli. In this respect, the complex multi-step phosphorelay mechanism seems to be disadvantageous, but there are several unorthodox two-component systems in E. coli. In this paper, we investigate the reason why such unorthodox two-component systems are present in E. coli. For this purpose, we have developed simplified mathematical models of both orthodox and unorthodox two-component systems and analyzed their dynamical characteristics through extensive computer simulations. We have finally revealed that the unorthodox two-component systems realize ultrasensitive responses to external stimuli and also more robust responses to noises than the orthodox two-component systems. PMID- 17112786 TI - HIV-1 immunopathogenesis: how good interferon turns bad. AB - The hallmark of acquired immunodeficiency syndrome (AIDS) is the progressive loss of CD4+ T cells that results from infection with human immunodeficiency virus type-1 (HIV-1). Despite 25 years of AIDS research, questions remain concerning the mechanisms responsible for HIV-induced CD4+ T cell depletion. Here we briefly review the in vitro and in vivo literature concerning the protective role of interferon-alpha (IFN-alpha) in HIV/AIDS. We then develop a laboratory- and clinically supported model of CD4+ T cell apoptosis in which either infectious or noninfectious HIV-1 induces the production of type I interferon by plasmacytoid dendritic cells (pDC). The interferon produced binds to its receptor on primary CD4+ T cells resulting in membrane expression of the TNF-related apoptosis inducing ligand (TRAIL) death molecule. The binding of infectious or noninfectious HIV-1 to CD4 on these T cells results in expression of the TRAIL death receptor 5 (DR5), leading to the selective death of HIV-exposed CD4+ T cells. PMID- 17112787 TI - Extra-long PCR, an identifier of DNA adducts in single nematodes (Caenorhabditis elegans). AB - DNA adducts are frequently caused by chemical induced changes in DNA. If mis repaired, they can lead to nucleotide substitutions, deletions or chromosomal rearrangements. Depending on adduct stereochemistry and properties of the DNA target, adducts can inhibit transcriptional mechanisms. Here we demonstrate how this phenomenon can be exploited to detect DNA adducts in individual nematodes (Caenorhabditis elegans). An extra-long (XL)-PCR (16,144 bp) target amplicon, the 11 exon spanning ced-1, could be amplified reliably from genomic lysate extracted from single nematodes. Amplification efficiency was assessed by means of a second, fully quantitative PCR. Following the normalization with an invariant control gene, adduct formation could be evaluated by the identification of XL-PCR amplifications that were, relative to the control gene, reduced or inhibited by >95%. No DNA adducts could be detected in C. elegans maintained under optimal growth conditions (no exposure controls) or nematodes exposed to 20 microg/g copper sulfate (exposure negative control). However, exposure to 5 mug/g benzo[a]pyrene induced a stark response, with 40% of nematodes displaying measurable DNA adducts. Similarly, adducts were identified in 10% of nematodes subjected to 3 microg/g fluoranthene or a mixture containing 0.5 microg/g benzo[a]pyrene and 1 microg/g fluoranthene. PMID- 17112788 TI - Synergistic actions of enalapril and tempol during chronic angiotensin II-induced hypertension. AB - Experiments were designed to test the hypothesis that antioxidant treatment would increase the anti-hypertensive actions of endogenous kinins during angiotensin converting enzyme (ACE) inhibition. Four groups of rats, all given angiotensin II (Ang II) for 2 weeks, were studied: 1) control, 2) enalapril, 3) tempol or 4) both tempol and enalapril. Ang II significantly increased systolic blood pressure (BP) when compared with the baseline (170+/-8 vs. 128+/-4 mm Hg, P<0.05). Neither enalapril nor tempol alone was able to attenuate the elevation in BP (165+/-7 and 164+/-6 mm Hg, respectively). In contrast, combined administration of tempol and enalapril prevented the increase in BP (137+/-5 mm Hg). Plasma 8-isoprostane increased in Ang II-infused rats when compared with control untreated rats (69+/ 14 vs. 23+/-0.5 pg/ml, P<0.05). Tempol alone or tempol plus enalapril significantly attenuated the increase in plasma 8-isoprostane (29+/-6 and 34+/-7 pg/ml, respectively). In additional experiments, we used the bradykinin B(2) antagonist, icatibant to determine if increased B(2) receptor contributes to the anti-hypertensive effect of combined tempol and enalapril in Ang II-infused rats. Icatibant decreased the ability of this combination to lower arterial pressure. Additionally, a significant increase in B(1) receptor protein expression in renal cortex of Ang II-infused rats was observed compared to control suggesting that bradykinin receptor activation could account for the effect of enalapril to enhance the actions of tempol. These data support the hypothesis that combined reduction of superoxide along with enhanced endogenous kinins may facilitate blood pressure lowering in Ang II hypertension. PMID- 17112789 TI - Combination therapy using aspirin-enhanced photodynamic selective drug delivery. AB - In photodynamic therapy (PDT), excitation of a drug by light leads to a cascade of biochemical processes that can cause closure of blood vessels. It has been observed clinically that significant short-term leakage from the irradiated vasculature can occur prior to vessel closure and blood flow stasis. In this paper we demonstrate in a chicken embryo model that this leakage can be significantly enhanced by the presence of the cyclo-oxygenase inhibitor, aspirin. We also observe that following this aspirin-enhanced leakage, blood vessels close as effectively as after PDT in the absence of aspirin. Consequently we propose that this PDT-induced aspirin-enhanced leakage can be used to locally deliver a drug for combination therapy. This is then demonstrated in the chicken embryo using Visudyne as a PDT agent in combination with aspirin and fluorescein isothiocyanate dextran 10 kDa as leakage indicator. The latter represents a hypothetical drug to be delivered in various kinds of combination therapy. Two examples of this procedure would be the photodynamic treatment of choroidal neovasculature associated with exudative age-related macular degeneracy (AMD) where local delivery of an anti-angiogenic or an anti-inflammatory drug has been shown to be effective, or PDT of cancer where local dosing of a chemotherapeutic drug may well increase the treatment efficacy. PMID- 17112790 TI - lyl-1 and tal-1/scl, two genes encoding closely related bHLH transcription factors, display highly overlapping expression patterns during cardiovascular and hematopoietic ontogeny. AB - The TAL-1/SCL and LYL-1 genes encode two closely related basic helix-loop-helix transcription factors involved in child T-acute lymphoblastic leukemia through chromosomal rearrangements and transcriptional deregulation. During ontogeny, Tal 1/SCL is required for hematopoietic cell generation, both in the yolk sac, where erythro-myeloid cells are first produced, then in the intra-embryonic compartment, where hematopoietic stem cells independently arise. We describe here the expression pattern of lyl-1 in mouse embryos from 7 to 14 days post coitus using in situ hybridization, as well as beta-Galactosidase (beta-Gal) expression in lyl-1-lacZ knock-in embryos, which express a C-terminally truncated Lyl-1 protein fused to the beta-Galactosidase (Lyl-1Delta/beta-Gal). In addition, we compare lyl-1 expression pattern with that of tal-1/scl. Similar to Tal-1/SCL, Lyl-1 mRNA expression occurs in the developing cardiovascular and hematopoietic systems. However, contrary to tal-1/scl, lyl-1 is not expressed in the developing nervous system. In lyl-1-lacZ knock-in heterozygous and homozygous embryos, beta Gal expression completely correlates with Lyl-1 mRNA expression in the intra embryonic compartment and is present: (1) in the developing hematopoietic system, precisely where hematopoietic stem cells emerge, and thereafter in the fetal liver; (2) in the developing vascular system; and (3) in the endocardium. In contrast, whereas Lyl-1 mRNA is expressed in yolk sac-derived endothelial and hematopoietic cells, Lyl-1Delta/beta-Gal is either absent or poorly expressed in these cell types, thus differing from Tal-1/SCL, which is highly expressed there at both mRNA and protein levels. PMID- 17112791 TI - Repair of alkylated DNA: recent advances. AB - Cytotoxic and mutagenic methylated bases in DNA can be generated by endogenous and environmental alkylating agents. Such damaged bases are removed by three distinct strategies. The abundant toxic lesion 3-methyladenine (3-alkyladenine) is excised by a specific DNA glycosylase that initiates a base excision-repair process. The toxic lesions 1-methyladenine and 3-methylcytosine are corrected by oxidative DNA demethylation catalyzed by DNA dioxygenases. These enzymes release the methyl moiety as formaldehyde, directly reversing the base damage. The third strategy involves the mutagenic and cytotoxic lesion O(6)-methylguanine which is also repaired by direct reversal but uses a different mechanism. Here, the methyl group is transferred from the lesion to a specific cysteine residue within the methyltransferase itself. In this review, we briefly describe endogenous alkylating agents and the extensively investigated DNA repair enzymes, mammalian 3-methyladenine-DNA glycosylase and O(6)-methylguanine-DNA methyltransferase. We provide a more detailed description of the structures and biochemical properties of the recently discovered DNA dioxygenases. PMID- 17112792 TI - The mechanics of base excision repair, and its relationship to aging and disease. AB - Base excision repair (BER) is the major pathway responsible for averting the mutagenic and cytotoxic effects of spontaneous hydrolytic, oxidative, and non enzymatic alkylation DNA damage. In particular, this pathway recognizes and repairs base modifications, such as uracil and 8-hydroxyguanine, as well as abasic sites and DNA single-strand breaks. In this review, we outline the basic mechanics of the BER process, and describe the potential association of this pathway with aging and age-related disease, namely cancer and neurodegeneration. PMID- 17112793 TI - Determination of citalopram and escitalopram together with their active main metabolites desmethyl(es-)citalopram in human serum by column-switching high performance liquid chromatography (HPLC) and spectrophotometric detection. AB - We established a method for automated quantitative analysis of (es-)citalopram and desmethyl(es-)citalopram in serum using column-switching high performance liquid chromatography (HPLC). For sample clean-up serum was injected onto a LiChrospher CN 20 microm precolumn using 8% acetonitrile in deionized water. Drugs were eluted by back-flush flow onto the analytical column (LiChrospher CN 5 microm) at a flow rate of 1.5 ml/min with phosphate buffer 8 mmol/l pH 6.4/acetonitrile (50/50, v/v). Haloperidol was used as internal standard. Analytes were detected by ultraviolet spectrophotometry at 210 nm. Detection limit of (es-)citalopram was 6 ng/ml. The method was found to be suitable for therapeutic drug monitoring of patients treated with citalopram or escitalopram. PMID- 17112794 TI - One-step immunochromatographic separation and ELISA quantification of glycyrrhizin from traditional Chinese medicines. AB - The bioactive constituent, glycyrrhizin or glycyrrhizic acid (GA), was purified from two traditional Chinese medicines (TCM), Shaoyao gancao tang and Dahuang gancao tang, and from crude extracts from licorice roots by means of immunoaffinity chromatography using anti-GA monoclonal antibody (MAb) and was quantified with an enzyme-linked immunosorbent assay (ELISA). Laboratory preparations included the synthesis of conjugate GA-human serum albumin (GA-HSA), the production of anti-GA-MAb, the optimization of the immunoaffinity column packed with the anti-GA-MAb coupled to hydrazide gel and the determination of the GA content in TCM and crude drugs from five different sources by ELISA and high performance liquid chromatography (HPLC). The experimental results reveal that the anti-GA-MAb coupled to Affi-Gel Hz gel results in a coupling efficiency of 95.2%, and the immunoaffinity chromatography gives a mean recovery of 97.6% of GA with a capacity of 33.5+/-2.40 microg/mL of immunoaffinity gel under the given conditions. The GA content of the crude extracts (ranging 74.8-114.6 microg/mg) from different sources by the ELISA method is much greater than that of the TCM (16.4-25.1 microg/mg) which is, in good agreement with the results of the HPLC method. Our report provides a rapid, reliable and sensitive approach for one-step separation and quantification of GA. PMID- 17112795 TI - Unravelling in vitro variables of major importance for the outcome of mass spectrometry-based serum proteomics. AB - The use of mass spectrometry (MS) for analysing low-molecular weight proteins and peptides from biological fluids has a great, yet not fully realized, potential for biomarker discovery. To prune MS-data as much as possible for non-relevant non-biological variation the development of standardized protocols for handling and processing the samples before MS and adjusting data after MS to compensate for method-induced variability are warranted. This calls for knowledge about how different variables contribute to MS-based proteome analyses. In addition, identification of the peptides involved in pre-analytical variation will be helpful in evaluating the clinical significance of predictive models derived from MS data. Using human sera, extraction by weak cation-exchange magnetic beads, and analysis by MALDI-TOF MS we here evaluated pre-analytical variation and identify peptides involved in this. The influences of humidity, temperature, and time for preparation of sera on spectral changes were evaluated. Also, the reproducibility of the methods and the effect of a baseline correction procedure were examined. Low temperatures, short handling times, and a baseline correction procedure minimize the contribution of artifacts to sample variability as observed by MS. The complement split product C3f and fragments thereof appear to be sensitive indicators of sample handling induced modifications. Other peptides that are indicative of such variability are fibrin and kininogen fragments. Using strict experimental guidelines as well as standardized sample collection procedures it is possible to obtain reproducible peak intensities and positions in serum mass profiling using magnetic bead-based fractionation and MALDI-TOF MS. PMID- 17112796 TI - Optimum conditions of autoclaving for hydrolysis of proteins and urinary peptides of prolyl and hydroxyprolyl residues and HPLC analysis. AB - A method for urinary peptide(s) and protein hydrolysis, involving autoclaving at 15psi (121 degrees C) for 60min, is described. Using three candidate proteins (bovine serum albumin, casein and gelatin) and urine specimens, the effect of autoclaving with respect to the optimum time required for hydrolysis under both acidic (6N HCl) and alkaline (6N KOH) conditions was studied. Recoveries of total amino acids from proteins and urine hydrolysate(s) suggest that complete hydrolysis of proteins and urinary peptides could be achieved by autoclaving for 30-60min instead of 16h of incubation at 110 degrees C. Further, stability of some of the individual amino acids was also studied. The observed differential stability of amino acids under acidic and alkaline conditions, as demonstrated in this study by HPLC analysis, makes it imperative to choose the appropriate hydrolytic condition while studying the composition of any given amino acids in urinary peptide(s)/protein hydrolysates. Further, the finding that both Pro and Hyp were stable under alkaline conditions of hydrolysis by autoclaving renders this method suitable for assaying these two amino acids from urine hydrolysates, hence its utility in the study of urinary peptide derived Hyp and Pro in bone/cartilage disorders. PMID- 17112797 TI - Colon cancer prevention in Italy: cost-effectiveness analysis with CT colonography and endoscopy. AB - BACKGROUND: Colorectal cancer (CRC) is a major cause of mortality in Italy. Although prevention of CRC is possible, its cost-effectiveness when applied to the Italian population is unknown. Recently, computerized tomographic colonography (CTC) has been proposed for CRC screening. AIM: To compare the efficacy and cost-effectiveness of CTC screening in a simulated Italian population with those of colonoscopy and flexible sigmoidoscopy (FS). METHODS: The cost-effectiveness of different screening strategies was compared using a Markov process computer model, in which in a hypothetical population of 100,000 50 year-olds were investigated by CTC, colonoscopy or FS every decade. Outcomes were projected to the Italian national level. RESULTS: CRC incidence reduction was calculated at 40.9%, 38.2%, and 31.8% with colonoscopy, CTC and FS, respectively. As compared to no screening, all screening programs were shown to be cost-saving, allowing a saving of 11 Euro, 17 Euro, and 48 Euro per person with colonoscopy, FS and CTC, respectively. FS appeared to be less cost-effective than CTC, whilst colonoscopy appeared to be an expensive option as compared to CTC. Undiscounted national expenditure was calculated to be 1,042,489,512 Euro, 1,093,268,285 Euro, and 1,198,783,428 Euro for FS, CTC and colonoscopy, respectively, as compared to 695,818,078 Euro without screening. CONCLUSION: CRC screening is cost-saving in Italy, irrespective of the technique applied. CTC appeared to be more cost-effective than FS, and it may also become a valid alternative to colonoscopy. PMID- 17112798 TI - Associations of vitamin D status with bone mineral density, bone turnover, bone loss and fracture risk in healthy postmenopausal women. The OFELY study. AB - INTRODUCTION: Vitamin D status is considered as an important determinant of bone health but supplementation trials with vitamin D(3) have yielded conflicting results. The aim of this study was to investigate the associations between serum 25-hydroxyvitamin D (25-OH D), bone turnover markers, bone mineral density (BMD), radius bone loss and incidence of fracture in postmenopausal women. METHODS: 669 postmenopausal women (mean age: 62.2 years) belonging to a population-based cohort were followed prospectively for a median of 11.2 years. At baseline, 25-OH D levels, BMD, bone turnover markers and clinical risk factors of osteoporosis were assessed. BMD loss at the radius was estimated by annual measurements of BMD and all incident fractures which occurred in 134 women were confirmed by radiographs. RESULTS: 73% and 35% of women had serum 25-OH D levels below 75 and 50 nmol/l which correspond respectively to the median and lowest optimal values recently proposed for fracture prevention. 11% of women had levels below 30 nmol/l. Serum 25-OH D correlated modestly with intact PTH (r(2)=0.023, p<0.0001), but not with bone turnover markers or BMD at the hip and radius after adjustment for age. When levels of 25-OH D were considered as a continuous variable, there was no significant association between 25-OH D levels and radius BMD loss or fracture risk. After adjustment for age, there was no significant difference in incidence of fracture, BMD, radius BMD loss, bone turnover markers, grip strength and the percentage of fallers in the previous year between women with 25-OH D levels below or above 75, 50 or 30 nmol/l. CONCLUSIONS: In a population of home dwelling healthy postmenopausal women with few of them with severe vitamin D deficiency, vitamin D status may not be an important determinant of bone health. PMID- 17112800 TI - ABCB1 gene polymorphisms are not associated with treatment outcome in elderly acute myeloid leukemia patients. AB - OBJECTIVES: The classical multidrug resistance (MDR) gene MDR1 (ABCB1) encodes for the drug efflux pump P-glycoprotein (P-gp). P-gp expression is an adverse prognostic factor for treatment outcome in acute myeloid leukemia (AML) and is more frequently observed in older patients. Single-nucleotide polymorphisms of the ABCB1 gene, C1236T, G2677T, and C3435T, have been associated with altered drug metabolism and treatment outcome. We prospectively determined these single nucleotide polymorphisms in AML blasts in a cohort of patients aged 60 years or older with AML and evaluated their relevance with regard to P-gp function and expression, ABCB1 messenger ribonucleic acid (mRNA) expression, and clinical outcome. METHODS: We have analyzed purified bone marrow-derived leukemic blasts, obtained at diagnosis, in 150 patients who were treated within a multicenter, randomized, phase 3 trial of elderly patients with AML. The significance of the allelic ABCB1 variants of C1236T, G2677T, and C3435T was evaluated with respect to P-gp expression and function in leukemic blasts and ABCB1 mRNA expression levels, and these values were correlated with treatment outcome. RESULTS: P-gp function and expression in leukemic blasts and ABCB1 mRNA levels in patients with AML did not vary significantly among any of the allelic variants of ABCB1. None of these allelic variations predicted a difference in complete response rate and survival endpoints. CONCLUSIONS: In AML patients aged 60 years or older, allelic ABCB1 variations of C1236T, G2677T, or C3435T are not associated with altered P gp function or with MDR1 expression at the transcriptional or translational level in leukemic blasts, and they do not significantly affect clinical prognosis. PMID- 17112801 TI - Transcriptional profiling of genes induced in the livers of patients treated with carbamazepine. AB - BACKGROUND: The antiepileptic drug carbamazepine (CBZ) is a potent inducer of human drug metabolism, resulting in serious interactions with many commonly prescribed drugs. The molecular mechanisms underlying this response are not well understood, however, and the spectrum of CBZ-inducible genes in human liver has not been thoroughly investigated. METHODS: The availability of liver ribonucleic acid from 2 epileptic patients treated with CBZ and from 7 control subjects enabled us to study the global induction response of drug-metabolizing enzymes, drug transporters, and nuclear receptors in vivo. RESULTS: Using expression profiling, we identified 64 significantly up-regulated transcripts but only 1 significantly down-regulated transcript (SLC22A5). We confirmed the induction of several genes that previously have been shown to be inducible by drugs in vitro, including multiple cytochrome P450 (CYP) genes in the CYP1A, CYP2A, CYP2B, CYP2C, and CYP3A subfamilies, as well as glutathione S-transferase A1, uridine diphosphate-glucuronosyltransferase 1As, the drug transporter ABCC2, and the nuclear receptors CAR (constitutive androstane receptor) and PXR (pregnane X receptor). Moreover, we identified a number of additional genes not previously known to be induced by CBZ, including CYP39A1, sulfotransferase 1A1, glutathione S-transferase Z1, and the drug transporters SLCO1A2, ABCG2, and ABCB7, as well as the glucocorticoid and aldosterone receptors. In transactivation studies in CV-1 cells, we demonstrated that both CBZ and its major metabolite, CBZ-10,11-epoxide, activate the nuclear receptor PXR in a concentration-dependent fashion and at therapeutic concentrations with 50% inhibitory concentration values of approximately 50 micromol/L. CONCLUSIONS: CBZ is a potent inducer of a broad spectrum of drug-metabolizing enzymes and drug transporters in the human liver, and these effects are mediated at least in part by activation of PXR. PMID- 17112802 TI - CYP2A6 genotype and the metabolism and disposition kinetics of nicotine. AB - BACKGROUND AND OBJECTIVE: The liver enzyme cytochrome P450 (CYP) 2A6 is primarily responsible for the metabolism of nicotine. Variants in the CYP2A6 gene have been associated with altered nicotine metabolism and with effects on smoking behavior. Our objective was to determine the relationship between variant CYP2A6 genotypes and the disposition and metabolism of nicotine administered intravenously. METHODS: Intravenous infusions of deuterium-labeled nicotine and cotinine were administered to 278 healthy twin volunteers, most of whom were white. They were genotyped for CYP2A6*1, CYP2A6*2, CYP2A6*4, CYP2A6*7, CYP2A6*8, CYP2A6*9, CYP2A6*10, and CYP2A6*12. RESULTS: On the basis of the fractional clearance of nicotine to cotinine and on the plasma ratio of 3'-hydroxycotinine to cotinine, both shown to be indicators of CYP2A6 enzymatic activity, subjects were classified into 3 groups. Group 1 included wild-type variant CYP2A6*1/*1 (n=215) and was assumed to have 100% activity. Group 2 included *1/*9 (n=21) and *1/*12 (n=12), which averaged about 80% of normal activity. Group 3 included *1/*2 (n=10), *1/*4 (n=2), *9/*12 (n=3), *9/*4 (n=2), and *9/*9 (n=3), which averaged about 50% of normal activity. The mean total plasma clearance of nicotine (+/-SD) was 18.8+/-6.0, 15.5+/-4.9, and 11.7+/-5.1 mL.min-1.kg-1 in groups 1, 2, and 3, respectively, and group 1 had significantly faster clearance than group 2 (P<.05) and group 3 (P<.01). Overall, groups 2 and 3 also had lower total clearance of cotinine, had longer half-lives for nicotine and cotinine, and excreted in the urine a greater fraction of the nicotine dose as unchanged nicotine and nicotine glucuronide and excreted less as 3'-hydroxycotinine compared with group 1. CONCLUSIONS: We provide novel pharmacokinetic and metabolic data on nicotine after systemic dosing in relation to common CYP2A6 genotypes. Our data will enhance the interpretation of CYP2A6 genotypic data as used in association studies of smoking behavior and its health consequences. PMID- 17112803 TI - High-dose methotrexate in pediatric acute lymphoblastic leukemia: impact of ABCC2 polymorphisms on plasma concentrations. AB - OBJECTIVE: The adenosine triphosphate-binding cassette (ABC) class transporter ABCC2 (MRP2 [multidrug resistance related protein 2] or cMOAT [canalicular multispecific organic anion transporter]) is involved in the cellular outward transport and elimination of methotrexate. We hypothesized that common genetic variations may contribute to the variability of high-dose methotrexate pharmacokinetics. METHODS: Polymorphisms in all 32 exons of the ABCC2 gene were analyzed in a reference group of 59 healthy white subjects by polymerase chain reaction, single-strand conformation polymorphism, and sequencing. Subsequently, we assessed the association of polymorphisms with the methotrexate plasma concentrations in 44 pediatric patients with acute lymphoblastic leukemia (ALL) (29 male and 15 female patients; mean age, 6.8+/-4.8 years). Patients received 4 cycles of 5000 mg/m2 body surface area according to the ALL-Berlin-Frankfurt Muenster (BFM) 95 or ALL-BFM 2000 protocol. RESULTS: In the reference group we detected 8 frequent single-nucleotide polymorphisms. Five of these were in complete linkage disequilibrium. Overall, 5 new polymorphisms are described. The genotype distribution of the patient cohort was not significantly different from the reference collective. The mean plasma methotrexate area under the curve from 36 to 48 hours after the start of the infusion was significantly 2-fold higher in female patients carrying at least 1 -24T allele as compared with all other patients (14.2+/-12.8 h.micromol/L versus 6.9+/-4.2 h.micromol/L, P<.001). The risk to have 2 or more cycles necessitating an intensification of folinate rescue was 9-fold (95% confidence interval, 1.8- to 44-fold) in female patients carrying at least 1 T allele (P=.0067). CONCLUSION: The data suggest a hitherto unknown gender-specific impact of the -24C>T ABCC2 gene polymorphism on high-dose methotrexate pharmacokinetics. Whereas a nonfunctional MRP2 variant has been described in a patient with severe impairment of methotrexate excretion, our study is the first to suggest that a frequent ABCC2 polymorphism contributes to variability of methotrexate kinetics. PMID- 17112804 TI - Disposition and sterol-lowering effect of ezetimibe are influenced by single-dose coadministration of rifampin, an inhibitor of multidrug transport proteins. AB - BACKGROUND AND AIMS: The disposition and sterol-lowering effect of ezetimibe are associated with long-lasting enterosystemic circulation, which is initiated by secretion of ezetimibe and its glucuronide via intestinal P-glycoprotein (P-gp) (ABCB1) and the multidrug resistance-associated protein 2 (MRP2) (ABCC2) into gut lumen. Hepatic uptake and secretion may contribute to recycling. To obtain deeper insight into the intestinal and hepatic processes, the disposition of ezetimibe was studied in the presence of rifampin (INN, rifampicin), a modulator of P-gp, MRP2, and hepatic organic anion (uptake) transporting polypeptides (OATPs) (SLCOs). METHODS: The disposition of ezetimibe (20 mg orally) alone and after coadministration of rifampin (600 mg orally) was measured in a crossover study of 8 healthy subjects with the SLCO1B1 *1a/*1a genotype. Concentrations of ezetimibe and its glucuronide in serum, urine, and feces, as well as cholesterol, lathosterol, and the plant sterols campesterol and sitosterol in serum, were quantified by use of liquid chromatography and gas chromatography with mass spectrometric detection. RESULTS: After rifampin administration, the maximum serum concentrations of ezetimibe and its glucuronide were significantly elevated (12.0+/-4.20 ng/mL versus 4.67+/-2.72 ng/mL, P=.017, and 282+/-73.8 ng/mL versus 107+/-35.3 ng/mL, P=.012, respectively). The area under the curve of ezetimibe was not affected (102+/-37.6 ng.h/mL versus 140+/-86.3 ng.h/mL, P=not significant), whereas that of the glucuronide was markedly increased (2150+/-687 ng.h/mL versus 1030+/-373 ng.h/mL, P=.012). Renal clearance remained unchanged. Fecal excretion of ezetimibe was markedly decreased (7.6+/-2.2 mg versus 10.4+/ 1.8 mg, P=.036), whereas renal excretion of the glucuronide was strongly elevated (4.8+/-1.9 mg versus 2.0+/-1.2 mg, P=.049) after coadministration. The onset of a significant sterol-lowering effect of ezetimibe was significantly shortened by rifampin coadministration. CONCLUSIONS: Coadministration of rifampin increases the maximum serum concentrations of ezetimibe but reduces its enterosystemic recycling, most likely by inhibition of the secretion of ezetimibe and its glucuronide via P-gp and MRP2. PMID- 17112805 TI - Impact of P-glycoprotein on clopidogrel absorption. AB - OBJECTIVE: The antiplatelet activity of clopidogrel is characterized by considerable interindividual differences. Variable intestinal absorption is suggested to contribute to the inconsistencies in response. We tested the hypothesis that the intestinal efflux transporter P-glycoprotein (P-gp) limits the oral bioavailability of clopidogrel and that variance in the MDR1 gene encoding P-gp predicts absorption variability. METHODS AND RESULTS: P-gp-mediated transport of clopidogrel was assessed by transflux, influx, and efflux experiments by use of Caco-2 cells. Inhibition of P-gp activity by different modulators increased the absorptive clopidogrel flux across Caco-2 monolayers from 0.51+/-0.19 pmol/cm2 (mean+/-SD) at baseline by a maximum of 5- to 9-fold (P<.001) and the intracellular accumulation from 0.99+/-0.11 pmol/mg protein by a maximum of 2.5-fold (P<.001) in response to 1-micromol/L clopidogrel and decreased clopidogrel efflux to the level of passive diffusion. In 60 patients with coronary artery disease who underwent percutaneous coronary intervention, the peak plasma concentration (Cmax) and the total area under the plasma concentration-time curve (AUC) of clopidogrel and its active metabolite after a single oral loading dose of 300, 600, or 900 mg were tested for correlation with the MDR1 genotype. In the 300-mg and 600-mg groups (but not in the 900-mg group) Cmax and AUC values were lower in subjects homozygous for the MDR1 3435T variant compared with subjects with the 3435C/T and 3435C/C genotypes. After the 600-mg loading dose, Cmax values (mean+/-SD) of clopidogrel and its active metabolite in 3435T/T carriers were 13.3+/-5.2 ng/mL and 2.5+/-1.2 ng/mL, respectively, compared with 49.7+/-41.6 ng/mL (P=.001) and 6.6+/-3.6 ng/mL (P=.011), respectively, in 3435C/T and 3435C/C carriers; AUC values were 1502+/-463 ng/mLxmin for clopidogrel and 209+/-99 ng/mL x min for its active metabolite in 3435T/T carriers compared with 7057+/-5443 ng/mLxmin (P=.0006) and 744+/-541 ng/mLxmin (P=.011), respectively, in 3435C/T and 3435C/C carriers. CONCLUSIONS: Clopidogrel absorption and thereby active metabolite formation are diminished by P-gp-mediated efflux and are influenced by the MDR1 C3435T genotype. PMID- 17112806 TI - Voriconazole, but not terbinafine, markedly reduces alfentanil clearance and prolongs its half-life. AB - OBJECTIVE: Alfentanil is a short-acting synthetic opioid analgesic, which is extensively metabolized, mainly by hepatic cytochrome P450 (CYP) 3A enzymes. Concomitant administration of alfentanil and CYP3A inhibitors may lead to clinically important drug interactions. We investigated the possible interactions between alfentanil and orally administered voriconazole and terbinafine. METHODS: A randomized crossover study design in 3 phases was used. Twelve healthy volunteers were given 20 microg/kg intravenous alfentanil without pretreatment (control), after oral voriconazole administration (400 mg twice on the first day and 200 mg twice on the second day), or after oral terbinafine administration (250 mg once daily for 3 days). Plasma concentrations of alfentanil were measured for 10 hours, and the pharmacokinetic parameters were calculated by use of noncompartmental methods. RESULTS: Voriconazole decreased the mean plasma clearance of intravenous alfentanil by 85%, from the control value of 4.4+/-2.4 mL.min-1.kg-1 to 0.67+/-0.27 mL.min-1.kg-1 (P<.001), and prolonged its elimination half-life from 1.5+/-0.49 hours to 6.6+/-1.8 hours (P<.001). The area under the alfentanil plasma concentration-time curve was increased by 6-fold by voriconazole (P<.001). Terbinafine had no statistically significant effect on the pharmacokinetics of alfentanil. Alfentanil administration caused nausea in 5 volunteers and vomiting in 2. These side effects all occurred in volunteers in the voriconazole phase. CONCLUSION: Oral voriconazole, but not terbinafine, markedly inhibited the metabolism of alfentanil. Caution should be exercised when alfentanil is given to patients receiving voriconazole. It is reasonable to assume that patients receiving voriconazole require 70% to 90% less alfentanil for the maintenance of analgesia than patients who are not receiving concomitant CYP3A inhibitors. PMID- 17112807 TI - Rifampin induces alterations in mycophenolic acid glucuronidation and elimination: implications for drug exposure in renal allograft recipients. AB - BACKGROUND: Exposure to mycophenolic acid (MPA) and its main metabolites (MPA 7-O glucuronide [MPAG] and MPA acyl-glucuronide [AcMPAG]) is characterized by a large interindividual and intraindividual variability, resulting in part from variability in glucuronidation (via uridine diphosphate-glucuronosyltransferase isoforms) and excretion via multidrug resistance-associated protein 2 (MRP2). It can be hypothesized that drugs interfering with glucuronidation and excretion will alter (Ac)MPA(G) exposure. METHODS: This prospective, open-label, nonrandomized, controlled pharmacokinetic interaction study included 8 stable renal allograft recipients, all treated with mycophenolate mofetil. Rifampin (INN, rifampicin), administered once daily (600 mg/d) for 8 days, was used as the probe drug because of its known effects on both uridine diphosphate glucuronosyltransferase activity and MRP2 transport capacity. A 12-hour pharmacokinetic time-concentration profile was assessed before rifampin administration was started, and this was repeated on the last day of rifampin administration. Total and free MPA, MPAG, and AcMPAG concentrations in plasma and urine were measured by use of HPLC with tandem mass spectrometry detection. RESULTS: Total MPA area under the plasma concentration-time curve (AUC) from 0 to 12 hours decreased significantly after rifampin coadministration (17.5% decrease [95% confidence interval (CI), 5.18%-29.9%]; P=.0234). This was mainly a result of a decrease in total MPA AUC from 6 to 12 hours (32.9% decrease [95% CI, 15.4% 50.4%]; P=.0078), representing decreased enterohepatic recirculation. Free MPA AUC from 6 to 12 hours decreased significantly, by 22.4% (95% CI, 4.71%-49.5%; P=.0391). Total MPAG and AcMPAG AUC from 0 to 12 hours increased by 34.4% (95% CI, 13.5%-55.4%; P=.0156) and 193% (95% CI, 30.3%-355%; P=.0078) respectively. Urinary recovery of MPAG and AcMPAG increased significantly (P=.0078), but renal clearance of these glucuronides did not change after rifampin coadministration. CONCLUSION: This study demonstrates an interaction between mycophenolate mofetil and rifampin, which is a result of induction of MPA glucuronidation and possibly also rifampin-associated alterations in MRP2-mediated transport of MPAG and AcMPAG. This interaction should be taken into account when rifampin or other drugs influencing pregnane X receptor activity are coadministered with mycophenolate mofetil. PMID- 17112808 TI - Effect of grapefruit juice on cytochrome P450 2A6 and nicotine renal clearance. AB - BACKGROUND AND OBJECTIVE: Grapefruit juice is an inhibitor of the cytochrome P450 (CYP) 3A4 enzyme and transporters such as P-glycoprotein and organic anion transporting polypeptides, leading to clinically important interactions. Our objective was to study the effect of grapefruit juice on the pharmacokinetics of nicotine, which is primarily metabolized by the CYP2A6 enzyme. METHODS: Ten volunteers were given a 2-mg oral dose of deuterium-labeled nicotine on 3 occasions together with 1 L of water, full-strength grapefruit juice, or half strength grapefruit juice. Concentrations of nicotine and its metabolites were analyzed in plasma and urine for 8 hours. RESULTS: Grapefruit juice inhibited the formation of cotinine from nicotine (area under the plasma cotinine concentration time curve from 0 to 8 hours of 6807 min.ng/mL, 7805 min.ng/mL, and 8007 min.ng/mL for full-strength grapefruit juice, half-strength grapefruit juice, and water, respectively; repeated-measures ANOVA, P=.009). The time to peak plasma concentration of cotinine was delayed (216 minutes, 159 minutes, and 147 minutes, respectively; ANOVA, P=.011), and the peak plasma concentration was lower with grapefruit juice compared with water (18 ng/mL, 21 ng/mL, and 22 ng/mL, respectively; ANOVA, P=.010). Oral clearance, peak plasma concentration, and time to peak plasma concentration of nicotine were not affected. Grapefruit juice increased the renal clearance of nicotine (231 mL/min, 219 mL/min, and 123 mL/min, respectively; ANOVA, P=.045) and cotinine (19 mL/min, 14 mL/min, and 16 mL/min, respectively; ANOVA, P=.002). CONCLUSIONS: Grapefruit juice inhibits the metabolism of nicotine to cotinine, a pathway mediated by CYP2A6, and increases the renal clearance of nicotine and cotinine. Nicotine oral clearance is not affected by grapefruit juice because the inhibition of hepatic metabolism is offset by the increase in the renal clearance of nicotine. However, other compounds metabolized by CYP2A6, as well as other drugs excreted via renal clearance mechanisms similar to those of nicotine, may be susceptible to significant pharmacokinetic grapefruit juice interactions. PMID- 17112809 TI - Sex differences in CYP3A activity using intravenous and oral midazolam. AB - BACKGROUND: Studies examining sex differences in CYP3A activity have produced conflicting results. Our objective is to investigate whether sex differences exist in CYP3A activity as assessed by intravenous (IV) or oral midazolam pharmacokinetic analysis in healthy volunteers. METHODS: Data from 13 previous studies were used. A single dose of IV midazolam (0.025 mg/kg) was administered to 66 white adults (37 women and 29 men; mean age, 36.3+/-7.7 years). A single dose of oral midazolam, 0.075 mg/kg (5 studies), 0.15 mg/kg (1 study), or 5 mg (1 study), was administered to 72 adults (71 white and 1 Asian; 37 women and 35 men; mean age, 38.3+/-8.9 years). Pharmacokinetic parameters were determined via population methods by use of a nonparametric adaptive grid program and a 2 compartment IV and 1-compartment oral absorption model. The maximum a posteriori probability Bayesian method was used to estimate each subject's pharmacokinetic parameters. Monte Carlo simulation was performed to determine the probability distribution of the area under the concentration-time curves (AUCs). RESULTS: Women exhibited 11% higher mean weight-corrected total body midazolam clearance and 28% higher oral clearance compared with men (PA polymorphism affects splicing of a CYP2D6 minigene. PMID- 17112819 TI - The management of head and neck melanoma. PMID- 17112820 TI - Fetal listing for organ transplantation in Canada. AB - This article presents an ethical and legal examination of whether a fetus should be listed to receive a transplanted organ. To date, relatively little discussion of this question has found its way into either the clinical or ethics literature. This article is divided into four sections. The first section analyzes the most common reasons against fetal listing offered by the nonclinicians with whom the author spoke. The two reasons involve the legal concepts of rights and best interests. Pivotal ethical foundations of the Canadian health system are also discussed so as to help develop a compatible allocation process for Canadian transplant programs. The second section analyzes common concerns raised in the author's discussions with clinicians. The third section presents four cardiac transplant scenarios to illustrate how relevant and sequential criteria for deciding whether, and in what circumstances, an available heart can be defensibly allocated to a fetus. The last section summarizes the decision process that reflects the preceding sections' analysis. Recognizing that the four scenarios do not exhaust the likely situations wherein a fetus and an infant might qualify for the same organ, the article closes with a recommendation that it be considered a catalyst for further analysis. PMID- 17112821 TI - Assessing medical student knowledge, attitudes, and behaviors regarding organ donation. AB - PURPOSE: To measure medical students' knowledge of the central issues in organ donation and transplantation and to understand their perception of the extent of training they received prior to and during medical school. METHODS: A previously validated, 41-question instrument assessing organ donation, allocation, and transplantation knowledge was directly administered to 537 first- and second-year medical students attending one of three Ohio schools from January through April 2005. Students were also asked about their support for organ donation and the donation training they had received. RESULTS: Two hundred sixty four first-year and 236 second-year students responded (response rate = 93%). Few students to date received donation and transplantation training before (11%) or during (22%) medical school. Second-year students were more likely than first-year students to have received training during medical school (40% vs 6%, P < .001) and to have read articles regarding donation (24% vs 15%, P = .017). However, both first- and second-year medical students answered the majority of the knowledge questions incorrectly (43% vs 48%, P < .002). Knowledge regarding brain death was lower among medical students compared to a random sample of Ohio adults (P < .001). Donation coursework prior to or during medical school was significantly associated with an increased knowledge regarding donation (odds ratio [OR] = 2.01, P = .001) and knowing where to find answers to patients' questions regarding donation (OR = 2.76, P < .001). CONCLUSION: Medical students have significant gaps in knowledge regarding the organ donation and transplantation system. Donation and transplantation education is associated with improved knowledge in the area and comfort in knowing how to address patient questions. PMID- 17112822 TI - Training for doctors and nurses to deal with bereaved relatives after a sudden death: evaluation of the European Donor Hospital Education Programme (EDHEP) in Germany. AB - The sudden death of a person is one of the most extreme and painful experiences for the relatives, and doctors require special communication skills to deal adequately with the bereaved. The Dutch European Donor Hospital Education Programme was developed to train doctors and nurses in talking to the bereaved relatives and to make the donation request. In Germany, the one-day workshop has been adapted to the German language, law, and clinical practice. The evaluation data of 75 workshops and experiences of 760 participants (doctors, nurses, and psychologists) are reported. The framework of, methods used, and issues dealt with in the workshop were clearly appreciated by the great majority of the participants. Criticisms and suggestions were directed mainly at the duration of the workshop. One third of the participants pleaded for a 2-day workshop, two thirds for a refresher course half a year later. The main effects reported were that two thirds of the participants rated that relatives could be helped, talked with, and cared for in a better way. The workshop participants also reported that they themselves were better able to cope with the situation and were more inclined to take on the task following the workshop. Finally, the necessities and limits of psychosocial training for doctors and their staff are discussed. PMID- 17112823 TI - Effects of financial incentives on the intention to consent to organ donation: a questionnaire survey. AB - INTRODUCTION: Shortage of donor organs is a serious problem for transplantation medicine. One controversial suggestion to increase the number of organ donors is financial incentives for consent. The aim of this study was to test whether different forms and amounts of financial incentives were apt to increase the consent to organ donation. MATERIALS AND METHODS: Data were collected via questionnaires in urban and rural regions of Austria and randomly assigned to settings with three different amounts of financial incentives. The questionnaire was designed by using the theory of planned behaviour of Ajzen. Parents 69 mothers and 35 fathers; ages 25 to 65 years were evaluated for intention to consent to organ donation, perceived social norm, and positive/ negative aspects of organ donation without and with various financial incentives. RESULTS: The intention to consent to organ donation dropped highly significantly (Z = -7.556 P = .000) from the basic condition (M = 1.13; confidence interval [CIs] 0.78 to 1.51) to the condition with financial incentives (M = -1.58; CI, 1.96 to -1.15). No influence of the amount of financial incentive was observed. Highly significant differences were measured between both conditions for the social norm (Z = -5.638; P < .000) and the attitude toward organ donation (Z = -1.962; P < .05; Z = -2.104; P < .035). CONCLUSIONS: Financial incentives led to decreased consents and elicited strong rejections and negative reactions of the participants. Taking money for consent to organ donation seems to be a strict taboo for most people in Austrian society. PMID- 17112824 TI - Current knowledge and attitudes about organ donation and transplantation among Chinese university students. AB - Current attitudes toward organ donation among university students in mainland China and the differences in attitudes between Chinese students in mainland China versus overseas are unknown. To address these issues, we conducted a cross sectional survey using questionnaires among 922 Chinese undergraduates from mainland China and overseas regions of the world. Data were analyzed by descriptive statistics, Student t tests, chi-square tests, and a logistic regression analysis. We found that blood donors showed significantly better awareness of heart, liver, lung, skin, and tendon donation among commonly transplanted organs/tissues. As to the willingness for cadaveric organ donation, 61.3% of respondents consented, 8.5% objected, and 30.3% answered "not sure." The percentage holding an organ donor card was 15.7% among students from Hong Kong; 3.0%, mainland China; 2.8%, Macau; 2.6%, Taiwan, and 4.0%, other regions of the world. In a logistic regression analysis, female students (odds ratio [OR], 2.24; 95% confidence interval [CI], 1.35 to 3.72) and blood donors (OR, 1.91; 95% CI, 1.10 to 3.32) did, but age and study specialty (medical vs nonmedical) did not show significantly more positive attitudes toward cadaveric organ donation. Compared with students from mainland China, overseas Chinese students from various regions did not show significantly different attitudes toward cadaveric organ donation. In summary, blood donors among university students have a greater knowledge of transplantation and a more positive attitude toward organ donation. Since university students are an important source of blood donors in China, they will be a potential pool of organ donors in the future. PMID- 17112825 TI - Effects of ischemia-reperfusion injury on apolipoprotein M expression in the liver. AB - The present study investigated the expression pattern of apolipoprotein M (apoM) mRNA in a rat model of hepatic ischemia-reperfusion injury (IRI). Animals were ischemic for 1 hour followed by various reperfusion times. As expected, serum alanine aminotransferase levels were significantly increased under IRI, which indicated the severity of liver injury. Hepatic mRNA levels of HSP70, which is the most common characterized protein within the family of heat-shock proteins (HSP), were significantly increased after 0.5 to 3 hours of IRI. Plasma C reactive protein, high-density lipoprotein-cholesterol, and lipoprotein (a) levels were significantly increased after 1-hour ischemia followed by 0.5 to 3 hours of reperfusion. Interestingly, similar to HSP70, apoM mRNA levels in the liver were gradually increased after 0.5 to 3 hours of IRI, whereas it returned to a lower level after 6 or 24 hours of IRI, which indicated that hepatic apoM expression was significantly influenced by the acute phase of IRI. However, plasma apoM levels were not increased in parallel, even slightly decreasing after 0.5 or 1 hour of IRI. We concluded that apoM mRNA expression pattern, like HSP70, in the liver showed rapid, significant changes during hepatic local IRI. PMID- 17112826 TI - Beneficial effect of N-acetyl-cysteine on renal injury triggered by ischemia and reperfusion. AB - Reactive oxygen species are critical mediators of the early phase of ischemic (IR) injury. The contribution of antioxidants, such as N-acetyl-cysteine (NAC), in ameliorating the parenchymal lesions, inflammatory parameters, and functional variables in renal IR is still controversial. We studied the effect of NAC administration on renal injury induced by IR. Mice were subjected to renal pedicle occlusion and subsequent reperfusion for 24 or 120 hours. NAC was administered prior to surgery at two concentrations (40 or 300 mg/kg, i.p.). Renal function and acute tubular necrosis were assessed, as well as immune phenotyping of infiltrating cells, by flow cytometry. At 40 mg/dL of NAC, we did not observe any significant improvement in renal function (1.85 +/- 0.43 md/dL, P = .367) or tissue architecture (% of ATN: 2.51 +/- 0.27 mm, P = .852) compared to the controls (1.87 +/- 0.43 mg/dL and 3.12 +/- 0.34 mm, respectively). However, animals that received 300 mg/dL of NAC showed lower serum creatinine values (24 hours: 1.25 +/- 0.54 mg/dL) compared to controls (P = .009) and less extensive acute tubular necrosis (1.54 +/- 0.12 vs, P < .05). Treatment with 300 mg/dL of NAC decreased renal dendritic cell infiltration. The protective effect of NAC was better observed at high concentrations and early times. PMID- 17112827 TI - Ulinastatin attenuates lung ischemia-reperfusion injury in rats by inhibiting tumor necrosis factor alpha. AB - Ischemia-reperfusion (I/R) injury may influence graft function following transplantation. Ulinastatin, a urinary trypsin inhibitor has been shown to attenuate I/R injury in various organs such as intestine, heart, and kidney in animals. The present experiment was designed to evaluate the effect of pretreatment with ulinastatin on I/R-induced lung injury. METHODS: After establishing a constant left lung warm ischemia-reperfusion model in rats, 45 animals were randomly divided into three experimental groups: sham group (n = 15), IR group (n = 15), and ulinastatin (5000 U/kg pre-ischemia) + IR group (n = 15). The lung injury was evaluated by tissue myeloperoxidase activity, with simultaneous estimation of the serum concentration of TNFalpha. RESULTS: The ulinastatin-pretreated animals exhibited markedly decreased lung tissue myeloperoxidase activity (P < .05). Blood gas analysis demonstrated, that the treated animals had significantly ameliorated pulmonary oxygenation (P < .05). The serum concentration of TNF-alpha in the ulinastatin-pretreated group was markedly decreased compared with that of the I/R group (P < .05). CONCLUSIONS: Ulinastatin attenuated I/R-induced lung injury. This function is partly related to the capacity of the agent to inhibit myeloperoxidase activity in lung tissue and decrease systemic expression to TNF-alpha. PMID- 17112828 TI - Evaluation of sequential perfusion with Euro-Collins and Belzer solutions for pancreas preservation in rats. AB - OBJECTIVE: To investigate the effect of Euro-Collins and Belzer solutions in a sequential preservation of the pancreas. METHODS: Forty-five Wistar-EPM rats were divided into four groups, according to the solution used during preservation: (1) saline solution (SF): animals perfused and preserved with saline solution; (2) Euro-Collins group (C): animals perfused and preserved with Euro-Collins solution; (3) Belzer group (B): animals perfused and preserved with Belzer solution; (4) Euro-Collins/Belzer group (CB): animals perfused with equal parts of Euro-Collins and Belzer solutions sequentially and preserved with Belzer solution. After perfusion, the animals underwent pancreas resection and preservation with the respective substance at 4 degrees C. Amylase was measured in the preservation solution after 12, 24, 36, or 48 hours. Finally, the pancreas was analyzed histologically, and a statistical analysis was performed. RESULTS: Groups SF and C showed the highest amylase levels in the preservation solution during all periods. The levels were higher than in groups C and CB (P = .05). Amylase levels were similar in groups B and CB to 24 hours (P = .05). Histological analysis was significant for analysis of pancreas islet cells and edema. Groups B and CB were histologically similar (P = .001) and different from groups SF and C. CONCLUSION: Sequential perfusion using Euro-Collins and Belzer solutions was effective for pancreas preservation in rats up to 24 hours. PMID- 17112829 TI - Expression of antiapoptotic survivin and aven genes in rat heart tissue after traumatic brain injury. AB - We have recently shown that experimental traumatic brain injury (TBI) results in ultrastructural damage in heart tissue. The aim of this study was to determine the two antiapoptotic signals "survivin" and "aven" in rat heart tissue following TBI, and comparing the effects of erythropoietin (EPO) and methylprednisolone (MPS). Thirty-six Wistar-Albino female rats weighing 190 to 230 g were randomly allocated into six groups: group 1 underwent head trauma with no treatment; group 2 and group 3, head trauma and intraperitoneally delivered EPO (1000 IU/kg) and MPS (30 mg/kg), respectively; group 4 (vehicle), head trauma and intraperitoneal albumin (0.4 mL/rat); groups 5 and 6, control and sham-operated groups, respectively. Three-hundred g-cm impact trauma was produced by the method of weight-drop. Real-time quantitative polymerase chain reactions were used to estimate survivin and aven gene expression at the total RNA level. Both survivin and aven were higher among the treatment than the trauma group (P = .0006, .0001 and P = .0038, .0033, respectively). Comparing survivin and aven between EPO and MPS treatment groups showed no significance (P = .3027, .2171, respectively). Also, both survivin and aven were significantly higher among the treatment than the vehicle, the control, or the sham-operated groups. These findings suggested that both EPO and MPS may play important roles in the expression of antiapoptotic survivin and aven genes in heart tissue after TBI. PMID- 17112830 TI - Amelioration of graft ischemia-reperfusion injury by breviscapine in rat small bowel transplantation. AB - OBJECTIVE: We sought to evaluate the effects of breviscapine to ameliorate graft ischemia-reperfusion (I/R) injury in a rat small bowel transplantation model. METHODS: Thirty-six recipients were randomly divided into three groups (n = 12): operative controls, in which grafts were implanted immediately after harvesting; an I/R control group with grafts preserved in cold lactated Ringer's solution at 4 degrees C for 4 hours before transplantation; and a breviscapine group wherein the graft was treated in the same way as the I/R control group but breviscapine (25 mg/kg/d) was injected intraperitoneally into both the donors and the recipients for 3 days before the operation of and into the recipients after transplantation. We compared the pathological scores for I/R injury, apoptosis index, and content of malondialdehyde (MDA) in the graft. RESULTS: Breviscapine diminished the pathological change caused by I/R injury (breviscapine vs I/R control on 24 hours after operation, 1.50 +/- 0.55 vs 2.17 +/- 0.75; P < .05), decreased the apoptotic index (breviscapine vs I/R control at 24 hours after operation, 27.33 +/- 0.167 vs 73.83 +/- 0.077; P < .05), and reduced the graft tissue content of MDA (breviscapine vs I/R control on 24 hours after operation, 1.717 +/- 0.131 vs 3.167 +/- 0.196; P < .05). CONCLUSIONS: Breviscapine may protect the transplanted small intestine against I/R injury during transplantation in rats. PMID- 17112831 TI - Living donor kidney exchange for both ABO-incompatible and crossmatch positive donor-recipient combinations. AB - BACKGROUND: Strategies to decrease the wait time for kidney transplantation include the use of living donor kidneys. However, it is not always possible to donate directly, due to ABO blood type incompatibility or a positive crossmatch. Therefore, other options were explored, including a program for living donor kidney exchange. METHODS: All Dutch kidney transplantation centers agreed on a common donor kidney exchange protocol. The Dutch Transplantation Foundation is responsible for the allocation, crossmatches are centrally performed, and exchanges take place on an anonymous basis. Donors travel to the recipient centers. Surgical procedures are simultaneously scheduled. RESULTS: From January 2004, we registered in total 116 combinations consisting of blood type incompatible pairs (n = 62) and positive crossmatch pairs (n = 54). In eight match procedures we created 58 new donor-recipient combinations with negative crossmatches, including six triplets and 20 doublets. It proved to be significantly (P = .0014) less difficult to find a solution for the crossmatch positive combinations than for the blood type-incompatible combinations (67% vs 35%). CONCLUSION: The Dutch national living donor kidney exchange program resulted in a 50% success rate. Combining blood type-incompatible and crossmatch positive donor-recipient pairs in one program is a realistic option for all blood type combinations. PMID- 17112832 TI - Evaluation of proteinuria in healthy living kidney donor candidates. AB - BACKGROUND: Evaluation of living kidney donor candidates includes careful assessment for the presence or absence of kidney disease. Kidney donation has been considered to be at least relatively contraindicated if urinary total protein excretion is above the normal range. However, at the present time, there is no uniformly accepted level of urine total protein excretion that would exclude donation. Albumin excretion instead of total protein excretion as a criterion has not previously been evaluated. MATERIALS AND METHODS: This was a prospective observational study over a 3-year period in a single tertiary care center designed to assess current selection criteria for kidney donation with respect to urine total protein and albumin excretion. RESULTS: Twenty four percent (25 of 105) of healthy adult kidney donor candidates had elevated urinary total protein excretion rates (150 to 292 mg/24 h). Of these 105 candidates, 39 had simultaneous measurements of both urinary total protein and albumin. Although one-third (13/39) had elevated 24-hour urine total protein values, none had elevated urine albumin excretion. CONCLUSION: Measurement of albumin, the most common single protein found in urine, appears to be helpful in the evaluation of proteinuria in donor candidates. Many healthy adult kidney donor candidates have mildly elevated total protein excretion but normal albumin excretion. We believe that such patients should not be excluded from donation. PMID- 17112833 TI - Minimally invasive kidney transplantation: the first experience. AB - Minimally invasive procedures in recent years have gained widespread acceptance. Within the field of transplantation, laparoscopic living donor nephrectomy (LLDN), requiring a 6- to 10-cm incision, is now considered the optimal procedure. According to recent MEDLINE searches, no minimally invasive technique has been reported for kidney transplantation. Considering the rapid evolution of minimally invasive surgery during the last decade, there is little reason to believe that kidney transplantation in future will be excluded from this development. A novel minimally invasive technique for kidney transplantation (MIKT) is presented, restricted to a 7- to 9-cm incision and minimal dissection/tissue trauma. The kidney is meticulously prepared on the back table and placed in a fitted lateral, retroperitoneal pouch. All three anastomoses are performed with the kidney in its final "in situ" position, and ureter reimplantation is done by extravesical technique. Twenty-one patients have been transplanted by MIKT and followed in a prospective manner, along with a matched control group subjected to conventional kidney transplantation. Our results indicate that MIKT may be executed safely and quickly. Beneficial effects on postoperative pain/analgesia, recovery, and complications are suggested by this first MIKT experience. The technical solutions of MIKT are per se not unique. However, the incision is minimal and not larger than the one required for LLDN. Minimally invasive surgery seems particularly attractive in the immunosuppressed population, and even more so with the recent introduction of potent antiproliferative drugs. PMID- 17112834 TI - Detection of donor-specific anti-HLA class I and II antibodies using antibody monitoring system. AB - The antibody monitoring system (AMS, GTI Inc) is a solid enzyme-linked immunosorbent assay (ELISA) crossmatch test for the detection of immunoglobulin G (IgG) antibody to donor-specific solubilized HLA class I and class II antigens. The objective of this study was to compare the results of the AMS assay with donor-specific anti-HLA IgG antibodies (DS-HLA Abs), as determined by ELISA panel reactive antibody (PRA) and the flow cytometric crossmatch test (FCXM). A total of 107 sera were screened for the presence of HLA Abs by ELISA PRA (LAT-M, One Lambda Inc), the DS-HLA Abs were determined in 34 serum samples (31.8%) by an ELISA panel (LAT class I and class II, One-Lambda Inc) and FCXM. The FCXM and AMS assays were performed with matched lymphocytes from 56 donors. There was a significant degree of concordance (89.7%) between the two tests (P < .001). The sensitivity, specificity, positive predictive value, and negative predictive value of AMS assay to detect DS-HLA Abs was 88.2%, 94.5%, 88.2%, and 94.5%, respectively. The AMS is a simple, objective test, which has several advantages over the cell-based crossmatch test, such as elimination of non-HLA antibody reactivity, elimination of non-donor-specific antibody reactivity, no need for viable cells, and preparation of the donor's HLA antigens in advance. In summary, this study suggested that AMS may be useful as a supportive crossmatch test or as a monitoring test after transplantation to detect class I or class II DS-HLA Abs. PMID- 17112835 TI - Additional effect of hyperparathyroidism on inflammatory status and rHuEPO requirements in hemodialysis patients. AB - BACKGROUND: The aim of this study was to analyze the effects of elevated parathyroid hormone (iPTH) and C-reactive protein (CRP) on rHuEPO requirements and associated clinical and biochemical parameters of hemodialysis patients. METHODS: A total of 127 hemodialysis patients were included. Laboratory values from the previous 3 months (monthly measured CRP, iPTH, albumin, prealbumin, calcium, phosphorus, and hemoglobin) and clinical findings (rHuEPO requirements, iron supplements, Kt/V) were recorded retrospectively. Patients were subgrouped according to presence of hyperparathyroidism (mean iPTH > 350 pg/mL) and chronic inflammation (mean CRP > 8.5 mg/L) as group I (low iPTH, low CRP, n = 32), group II (high iPTH, low CRP, n = 32), group III (low iPTH, high CRP, n = 32), and group IV (high iPTH, high CRP, n = 31). RESULTS: We found that group IV had lowest hemoglobin (P < .0001, .0001, .01, respectively), albumin (P < .0001), prealbumin (P < .0001, .0001, .02, respectively), and highest rHuEPO requirements (P < .0001, .0001, .01, respectively) compared to other groups despite of similar iron indices. Group III also had lower albumin (P < .002, .0001, respectively), prealbumin (P < .001, .01, respectively), hemoglobin (P < .001, .005, respectively), but higher rHuEPO requirements (P < .01) compared to group I and group II. CONCLUSIONS: We propose that hyperparathyroidism increases rHuEPO requirements and aggravates the negative effects of chronic inflammation in hemodialysis patients. PMID- 17112836 TI - Immunoglobulin class (IgG, IgM) determination by dithiothreitol in sensitized kidney transplant candidates. AB - Immunoglobulin class plays an important role in the histocompatibility crossmatch test to predict hyperacute rejection in kidney transplantation. The existing data indicates that immunoglobulin (Ig) M antibodies, particularly when they are autoantibodies, are not deleterious to the renal allograft. We used the reducing agent dithiotreitol (DTT) to inactivate IgM but not IgG in the crossmatch assay to help sensitized patients have the chance for successful transplantation. In this descriptive study, 57 candidates for kidney transplantation with final positive crossmatches who had a history of panel-reactive antibody (PRA) greater than 30% were selected. Two of 57 patients had systemic lupus erythematous (SLE). The sera of patients were treated by DTT and then measured for cytotoxicity against donor lymphocytes and a panel of 12 cells using the complement-dependent cytotoxicity (CDC) method. Autocrossmatch was also performed to differentiate autoantibodies and alloantibodies by the CDC method. Of the 57 patients, six subjects (10.53%) had IgM and 51 patients (89.47%) IgG in their serum against donor lymphocytes. Also against panel cells, 39 of 57 patients (68.43%) had IgG, three patients (5.26%) had IgM, and 15 patients (26.31%) had both IgG and IgM antibodies. Autolymphocytotoxic antibodies were detected in 1.75% patients (1 of 57) who had SLE. According to our results, 5.26% of the patients who were IgM positive and IgG-negative for both crossmatch and PRA assays may experience successful kidney transplantation. PMID- 17112837 TI - Complication of hemodialysis graft: anastomotic pseudoaneurysm: a case report. AB - Pseudoaneurysm is a relatively rare complication of synthetic vascular prostheses in patients on hemodialysis treatment and comes from a needle puncture. Anastomotic pseudoaneurysm is also rare. We have reported a case in which an anastomotic pseudoaneurysm developed in the early period in a patient on hemodialysis treatment. PMID- 17112838 TI - Long-term outcome of renal transplantation in focal glomerulosclerosis. AB - INTRODUCTION: Focal segmental glomerulosclerosis (FSGS) has a tendency to recur frequently after kidney transplantation. We evaluated 12 cases to examine the incidence and long-term outcomes of recurrent FSGS. MATERIALS AND METHODS: Twelve patients with renal failure caused by FSGS received kidney allografts from living related donors. Tacrolimus or cyclosporine was used in combination with prednisolone and azathioprine or mycophenolate mofetil. RESULTS: The mean graft survival was 87.4 +/- 46.8 months. The graft survival rates in FSGS recipients were at 1 year, 100%; 5 years, 79.6%; 10 years, 68.2%. Two out of four recipients experienced graft loss due to chronic rejection. The other two out of four recipients with graft loss displayed severe proteinuria diagnosed as recurrence of FSGS. To treat recurrent FSGS, plasma exchange was partially effective to reduce proteinuria. CONCLUSION: Our incidence of recurrent FSGS is 16.7% with graft survivals at 5 and 10 years of 79.6% and 68.2%, respectively. The recurrence of FSGS happened after scheduled reductions in immunosuppressants. Careful observation is required with maintenance of immunosuppression in these patients. PMID- 17112839 TI - Bourneville-Pringle disease for kidney transplantation: a single-center experience. AB - The Bourneville-Pringle disease is an autosomal-dominant disease affecting the kidneys in about 60%, causing end-stage renal disease in about 10% of the cases. Among more than 2800 renal transplant recipients during the last 33 years, we had two patients with this original disease. A third patient who underwent bilateral nephrectomy is currently awaiting a graft. The first patient was diagnosed at the age of 20 years after a few episodes of retroperitoneal bleeding. At the age of 26 years her left kidney was removed after a rupture; it measured 7500 g, and the histology described angiomyolipomatosis. A year later she underwent a cadaveric kidney transplantation. Subsequently her right kidney was removed due to bleeding. She is currently 5 years posttransplant with stable kidney function and good health. Our second patient was nephrectomized at the age of 35 years and 38 years because of angiomyolipomatosis. She underwent a cadaveric kidney transplantation 7 years later. After 5 years of excellent kidney function and a year after her arteriovenous fistula was ligated her upperarm had to be amputated because of uncontrollable bleeding. After another 6 months, she displayed rapid progression of a jejunal tumor and during operation received 54 U of blood transfusion but died at the age of 49 years with a well-functioning graft. Our third patient consecutively underwent two nephrectomies because of angiomyolipomatosis of her kidneys at the ages of 25 and 28 years. She has two children with the same disease. In addition she carries Leyden mutation, which has caused deep venous thromboses and pulmonary emboli. She is currently on our waiting list for kidney transplantation. The Bourneville-Pringle disease is a rare indication for kidney transplantation; the prognosis of the patient is dependent on the original disease. PMID- 17112840 TI - Donor-reactive cytokine production after HLA-identical living related kidney transplantation: a protein-array analysis. AB - In the present pilot study, we investigated which proteins are produced after donor stimulation of peripheral blood mononuclear cells from recipients of HLA identical living related kidney transplant. We used a protein-array analysis to determine cytokines, chemokines, and growth factors in supernatant from donor stimulated mixed lymphocyte reaction cultures. Autologous cultures were considered to be negative controls. In 38 out of 42 proteins (90%), the donor response was higher compared with the autologous response. Therefore, we concluded that even after HLA-identical living related kidney transplantation we could measure a donor-reactive response, which we assumed was directed toward minor histocompatibility or non-HLA antigens. PMID- 17112841 TI - Microchimerism evaluation in recipients of living-related or unrelated deceased allograft renal transplants. AB - The presence of microchimerism in the peripheral blood of solid organ graft recipients has been associated with long-term solid organ acceptance, immunologic tolerance, and less aggressive immunosuppressive therapy. Molecular biology assays are among the most sensitive methods to detect microchimerism, primarily to evaluate Y chromosome sequences in females as indirect evidence of circulating male nucleated donor cells. We screened for the presence of the SRY sequence region in peripheral blood of 13 female recipients of male kidney grafts: 5 living-related and 8 deceased grafts. Only patients who received grafts from related living donors exhibited microchimerism. Five of 13 patients studied exhibited better graft outcomes, including the 4 who were positive for the SRY sequences. PMID- 17112842 TI - The importance of patient satisfaction and health-related quality of life after renal transplantation. AB - In the last decade, patient satisfaction and quality of life have been accepted as important components of quality of medical care. Therefore, the purpose of this study was to examine and compare the quality of life and satisfaction in renal transplant patients with end-stage renal disease. In this cross-sectional study, quality of life and patient satisfaction were analyzed in 356 renal transplant recipients, 104 hemodialysis patients, 186 peritoneal dialysis patients. All patient groups were asked to estimate their subjective quality of life and satisfaction by responding to Turkish adapted instruments PSQ III (patient satisfaction questionnaire), 15D (quality of life), and sociodemographic data. The patient satisfaction (PSQ III) and quality of life (15D) mean scores were apparently better among male renal transplant than peritoneal or hemodialysis patients (P < .05). Married transplant patients' health-related quality of life (F = 4.64; P < .05) and satisfaction (F = 4.57; P < .05) mean scores were significantly higher than single or widowed patients. Total scores on the PSQ III showed a significant positive correlation with the total scores of the 15D (r = .842; P < .0001). In this study, the benefits of kidney transplantation were documented by means of satisfaction and quality-of-life assessments. The Turkish adapted PSQ III 18-item scale has excellent reliability and validity. The measure may be valuable for use in clinical trials or routine patient care. PMID- 17112843 TI - Evaluation of performance factors affecting two formulations of cyclosporine in pediatric renal transplant patients. AB - Success of renal transplantation in children is largely due to improvements in immunosuppressant therapy since the introduction of calcineurin inhibitors. The aim of this study was to identify possible factors that result in formulation differences in the exposure of pediatric patients to cyclosporine (CsA). We examined the handling of the two major formulations of CsA in a group of pediatric renal transplant recipients. The pharmacokinetic profiles of both formulations were assessed, and the data stratified to assess the effects of age, gender, time posttransplant, and other concomitant drug therapy on the two CsA formulations. The microemulsified formulation (MEC) enhanced bioavailability compared to the older oil-based formulation (CYA), especially at C2, with more predictable and consistent absorption in children. This higher bioavailability allowed a 15% reduction of dosing to achieve equal drug exposure. The concentration achieved by MEC at C2 demonstrated a much higher correlation with area under the concentration curve (AUC) than the concentration at C0. In the case of CYA a strong correlation was obtained between AUC and the concentrations obtained at both C0 and C2. Calcium channel blockers increased AUC(0-8) for both CsA formulations. Norfloxacin and pravastatin cotreatment had no effect on either of the CsA formulations. In contrast, the bioavailability of CsA was increased in boys using MEC formulation but this gender-based difference was absent during the use of CYA. This suggests that caution is required for introduction of new formulations of drugs to pediatric patients to evaluate differential effects of age, gender, and concomitant drug therapy. PMID- 17112844 TI - Use of sirolimus to facilitate steroid withdrawal from a cyclosporine regimen. AB - OBJECTIVE: Chronic steroid therapy has been associated with many comorbidities of transplant immunosuppression. Because sirolimus blocks one of the sites affected by steroids, we sought to examine whether substituting this drug mitigated these toxicities. METHODS: We used intent-to-treat methodology to compare the clinical outcomes and laboratory results between 30 renal transplant recipients converted from steroids to sirolimus with a cohort of demographically matched subjects who were transplanted concurrently with the study group and maintained on steroids. All patients received ongoing cyclosporine-based immunosuppression. To compensate for pharmacokinetic interactions with sirolimus, the cyclosporine exposure was markedly reduced in the study group. Statistical comparisons utilized analysis of variance, chi-square tests, and log rank evaluation of Kaplan-Meier curves. RESULTS: Conversion from prednisone to sirolimus was accomplished without difficulty in 27 of 30 patients. Treatment failures among the converted patients were due to chronic allograft nephropathy (n = 1), recurrence of original disease (n = 1), or chronic rejection (n = 2). By intent-to-treat analysis, the outcomes were similar in the study versus concurrent control groups. Laboratory values showed triglyceridemia as an adverse reaction to sirolimus, and reduced leukocytes, to steroid withdrawal. The observed clinical benefits solely reflected the markedly reduced cyclosporine exposure. Based on responses to a questionnaire administered prior to versus 12 and 24 months after steroid withdrawal, several domains revealed improvements in subjective complaints. CONCLUSION: Conversion from prednisone to sirolimus in combination with cyclosporine was easily accomplished in most renal transplant recipients. Although a 2-year follow-up failed to reveal objective benefits of the maneuver (other than those consequent to reduced cyclosporine exposure), most patients reported a subjectively improved health status. PMID- 17112845 TI - MDR1 C3435T polymorphisms correlate with cyclosporine levels in de novo renal recipients. AB - BACKGROUND: Single nucleotide polymorphisms (SNPs) in the multidrug resistance (MDR1) gene correlate with the intestinal function of P-glycoprotein (PGP). PGP serves as a hydrophobic export pump that extrudes cyclosporine (CsA) across the luminal membrane thus preventing CsA absorption. These genetic variants may predict CsA exposure levels in the early posttransplantation period. METHODS: CsA absorption profiles were established in 75 renal transplant patients using total daily dose and body weight adjusted 4-hour area under the time-concentration curve, AUC(0-4)/mg dose/kg body weight, on posttransplant day 3. These patients were subsequently genotyped for C3435T and G2677T polymorphisms using real-time polymerase chain reaction. An analysis was conducted to assess the independent impact of C3435T and G2677T SNPs on CsA bioavailability. RESULTS: C3435T polymorphisms were found to be an independent predictor of CsA AUC(0-4)/mg dose/kg levels on postoperative day 3. An inverse correlation was found between the number of T alleles and AUC values such that every T allele was associated with an approximate 15% decrement in AUC(0-4)/mg dose/kg (P = .034). A similar nonsignificant trend was observed for G2677T polymorphisms. CONCLUSIONS: MDR1 SNPs are correlated with CsA exposure in the early post-transplant period. Polymorphisms, in conjunction with other criteria, may become a useful tool to optimize initial drug dosing in renal transplantation. PMID- 17112846 TI - Tacrolimus dosing in Chinese renal transplant patients is related to MDR1 gene C3435T polymorphisms. AB - Tacrolimus is an immunosuppressive drug with narrow therapeutic range and wide interindividual variations in its pharmacokinetics. P-glycoprotein (P-gp) plays an important role in the absorption metabolism of tacrolimus. The polymorphism C3435T of MDR1, the gene coding P-gp, may influence the expression and activity of P-gp. The aim of this study was to evaluate whether C3435T polymorphism was associated with the tacrolimus concentration/dose ratio. Sixty-six Chinese renal transplant patients enrolled in this study were surveyed for body weight and dosage and concentration of tacrolimus as well as MDR1 genotype by polymerase chain reaction followed by restriction fragment length polymorphism analysis. The results showed a significant association between tacrolimus levels per dose mg/kg/d and MDR1 gene C3435T polymorphism (P < .05). The CC patients displayed a lower tacrolimus level per dose than CT/TT patients. Pharmacogenetic methods might be employed prospectively to help dose selection and to individualize immunosuppressive therapy. PMID- 17112847 TI - Effective immunoprophylaxis with basiliximab plus triple therapy in renal transplantation: five-year single-center experience. AB - We studied prospectively the efficacy and safety of basiliximab combined with triple immunosuppression in adult recipients of > or = 1 HLA-mismatched deceased donor renal grafts. All studied patients received equal immunosuppressive drugs: 20 mg infusion of basiliximab on day 0 and on day 4, cyclosporine microemulsion (Neoral), mycophenolate mofetil, and methylprednisolone. An analysis of 1-year data assessed the incidence of acute rejection episodes, safety of this therapy, renal graft function, and patient and graft survivals. One hundred seventy-two patients were studied. The HLA-antigen mismatches were 2.9 +/- 0.9 (mean +/- SD), and the cold ischemia time was 22.0 +/- 7.5 hours. Fifty-three (31.5%) patients experienced delayed graft function. At 12 months, 5 (3.0%) patients experienced acute rejection. Six renal grafts were lost, but not from rejection. Two patients died. Sixty-six infections required treatment in the hospital. One carcinoma of cervix (in situ) and two basal cell carcinomas of skin were detected. Hypersensitivity reactions and cytokine-release syndrome were not observed. At 12 months, serum creatinine was significantly higher (119 +/- 46 micromol/L; P < .001) in patients with delayed graft function than in patients with immediate graft function (99 +/- 26 micromol/L). Patient and graft survivals were 98.8% and 97.1%, respectively. Basiliximab combined with this triple therapy was an efficient and safe immunosuppression strategy, demonstrated with very low incidence of acute rejections, an acceptable adverse event profile, excellent graft function, and high short-term survival rates in adult recipients of deceased donor renal transplant. PMID- 17112848 TI - Conversion to enteric-coated mycophenolate sodium from various doses of mycophenolate mofetil: results of a prospective international multicenter trial in maintenance renal transplant patients receiving cyclosporine. AB - Conversion from mycophenolate mofetil (MMF, CellCept) to enteric-coated mycophenolate sodium (EC-MPS, myfortic) is safe and effective in renal transplant patients treated with the standard dose of 2 g MMF. In this 6-month, international, multicenter, open-label, single-arm trial, a large cohort of maintenance renal transplant patients receiving different doses of MMF were converted under normal clinical conditions to equimolar doses of EC-MPS. Mean calculated creatinine clearance remained stable from the time of study entry (59.6 +/- 19.7 mL/min) to the end of the study (58.3 +/- 19.8 mL/min). Adverse events were reported by 152 patients (67%), with gastrointestinal complications being observed in 45 patients (20%). Thirty-three patients (15%) experienced adverse events or infections with a suspected relation to EC-MPS, including one case of anemia and two cases of leukopenia. Eleven patients (4.9%) required a reduction in EC-MPS dose and seven patients (3.1%) permanently discontinued EC MPS owing to adverse events. At month 6 after conversion, five patients (2.2%) experienced biopsy-proven acute rejection. There were no graft losses or deaths. These data support earlier findings that stable maintenance renal transplant patients receiving MMF with cyclosporine with or without corticosteroids can be converted to EC-MPS with no compromise in efficacy and tolerability, and no adverse effect on renal function. PMID- 17112849 TI - Tolerability of enteric-coated mycophenolate sodium to 1 year in combination with cyclosporine and corticosteroids in renal transplant recipients. AB - Enteric-coated mycophenolate sodium (EC-MPS) is therapeutically equivalent to mycophenolate mofetil, but delays release of mycophenolic acid until it reaches the small intestine. De novo renal transplant patients taking part in a 12-month, multicenter, randomized study received cyclosporine microemulsion (CsA-ME, early or delayed to day 6), EC-MPS, steroids, and interleukin-2 antagonist induction. Tolerability data relating to EC-MPS are reported. Ninety-seven patients were randomized to early CsA-ME and 100 patients to delayed CsA-ME. Median daily dose of EC-MPS was 1440 mg at all time points throughout the 12-month period. The most frequently reported adverse events were constipation, anemia, urinary tract infection, abdominal pain, leukopenia, and cytomegalovirus infection; there were four malignancies. Fifty patients (24.6%) discontinued EC-MPS prematurely by 12 months, including 42 patients (84%) who discontinued owing to adverse events. No patient discontinued treatment because of gastrointestinal adverse events. Two thirds of patients (137 [67.5%]) maintained full EC-MPS dose throughout the 12 month study and did not require any dose reduction or dose interruption. EC-MPS is well tolerated in de novo renal transplant recipients when administered in combination with CsA-ME and steroids, with low rates of dose reductions or interruptions. Gastrointestinal adverse events were responsible for dose reduction or interruption in only 5% of patients. PMID- 17112850 TI - Morbidity rates with reduced mycophenolate mofetil dosage: a single-center experience. AB - OBJECTIVE: Mycophenolate Mofetil (MMF) is a key component of available immunosuppressant regimens for transplantation. Traditionally a dose of 2 g/d is utilized, with the incidence of side effects, most notably gastrointestinal, being well known to transplant teams. The aim of this study was to assess the incidence of side effects and rejection rates with reduced doses compared to standard regimens of MMF. METHODS: Case notes of 117 renal transplants performed from 2002 to 2004 were reviewed retrospectively for the incidence of side effects and rejection rates for MMF doses of 1 g twice a day (n = 86) versus 500 mg twice a day (n = 27). RESULTS: All patients received MMF, steroids, and a calcineurin inhibitor or sirolimus. The incidence of side effects was higher at 1 g twice a day. Seventy-nine percent of the patients required dose reductions due to side effects. The incidence of acute rejection episodes was similar in both groups. CONCLUSION: The balance between adequate immunosuppression, side effects, and the risk of rejection is a constant dilemma faced by transplant surgeons. This study showed a reduced side effect profile with similar rejection rates when using a lower-dose regimen of MMF. This may have implications for future immunosuppressive protocols. PMID- 17112851 TI - A randomized trial comparing two corticosteroid regimens combined with mycophenolate mofetil and cyclosporine for prevention of acute renal allograft rejection. AB - INTRODUCTION: The launching of mycophenolate mofetil (MMF) has reduced the incidence of acute rejection episodes. We sought to evaluate the efficacy of decreasing the steroid dose. MATERIALS AND METHODS: This was a quasiexperimental, randomized, prospective trial. We enrolled 150 patients who received de novo renal transplantations from living or cadaveric donors, fulfilling the screening criteria. Patients were randomized to one of the following two arms: (A) MMF at a 2 g/d dose, cyclosporine (CsA) at a dose necessary to achieve target levels, and corticosteroids at the usual doses; (B) MMF at a 2 g/d dose, CsA at a dose necessary to achieve target levels, and corticosteroids at doses 50% lower than those of group A. RESULTS: Group A included 72 (48%) and group B, 78 patients (52%). There were no differences among the variables: leukopenia occurred in 11 patients in group A, and five patients in group B. Complications occurred in 67.4% (56) of group A, but only 32.6% (27) were related to infections. One case of urinary infection occurred in group B, while six occurred in group A. There was one case of acute rejection in group A, and none in group B. One graft loss occurred in group A. There were no differences in the remaining variables under study. DISCUSSION: The results showed an increased complication rate related to receiving usual steroid doses. There was no increase in acute rejection episodes among patients receiving 50% of the usual steroid dose. PMID- 17112852 TI - In vitro monitoring of in vivo development of human anti-thymoglobulin antibodies by ELISA. AB - Thymoglobulin (rATG), polyclonal immunoglobulin, is prepared from rabbits immunized with human thymocytes. It is effective in prevention and treatment of renal allograft rejection. Human antibodies against antilymphocyte preparations can reduce efficacy by accelerating drug clearance or by inducing serum sickness. We developed an enzyme-linked immunosorbent assay (ELISA) to study posttreatment development of anti-rATG. In an Institutional Review Board-approved trial, we tested 101 allograft recipients for anti-rATG antibodies. Patients received rATG intravenously at 1.25 to 2.0 mg/kg/d for 2 to 14 days. Serum samples were obtained pretreatment and at weeks 1, 2, 4, 6, and months 3 and 6 post-rATG. ELISA plates were coated with rATG (10 microg/mL). Samples were diluted 1:100 and tested in quadruplicate. Positive samples were titrated. Horseradish peroxidase conjugated (HRPO) affinity-purified goat anti-human immunoglobulin G (H&L) antibody reacted with bound human antibody. A chromagenic substrate for HRPO was added and optical density (OD, 490 nm) was read. An OD of twice the negative control was considered positive. Mean ODs of negative and positive controls were 0.113 +/- 0.030 and 1.042 +/- 0.196, respectively. Ten patients had detectable anti-rATG before rATG administration (1:100). Thirty-five of 101 patients (35%) developed anti-rATG antibody. Patients showed an initial positive anti-rATG antibody from days 8 to 59 after infusion and titers from 1:100 to 1:4000. In spite of rATG's postulated anti-B-cell activity, this study confirms that rATG induces sensitization at a frequency and titer seen with other xenogeneic antilymphocyte antibodies. Formation of such antixenoantibodies can have a negative impact on treatment response and hence warrant monitoring. PMID- 17112853 TI - Conversion from azathioprine to mycophenolate mofetil followed by calcineurin inhibitor minimization or elimination in patients with chronic allograft dysfunction. AB - The purpose of this study was to evaluate the effects of the conversion from azathioprine (AZA) to mycophenolate mofetil (MMF) followed by calcineurin inhibitor (CNI) elimination or minimization in patients with progressive chronic allograft dysfunction (CAD). METHODS: Between November 6, 1999 and February 12, 2003, 169 patients receiving CNI/AZA/prednisone (153 CsA; 14 tacrolimus) were included in this study. Demographics, immunosuppression, graft function, hematology, and biochemistry were obtained before (-6, -3, and -1 month) and 1, 3, 6, 9 and 12 months after and at last follow-up visit after conversion. RESULTS: Mean age was 34 +/- 12 years, 66% males, 51% Caucasian, and 72% living allograft recipients. Mean follow-up times before and after conversion were 32.4 and 19.4 months; 10 patients completed 3 years of follow-up. CNI elimination was performed in 39% and minimization in 61% of patients. Overall there was significantly improved graft function at 1 year after conversion (2.6 +/- 1.0 vs 2.1 +/- 0.6 mg/dL, P = .038). The slopes of the regression lines of 1/Cr vs time were significantly improved from preconversion to after conversion (-0.026 vs +0.007 mg(-1)/dL per day(-1), P = .001). There was a significant decrease in mean systolic (141 +/- 21 vs 135 +/- 22 mm Hg, P = .015) and diastolic (89 +/- 15 vs 84 +/- 14 mm Hg, P = .005) blood pressure values at 1 year. There were four episodes of acute rejection (Banff IA) treated with steroids. Three years after conversion, patient and graft survivals were 95% and 79%, respectively. One patient developed posttransplant lymphoproliferative disease. CONCLUSION: Among patients with CAD, conversion from AZA to MMF followed by CNI minimization or elimination was a safe and effective strategy to preserve or improve graft function. PMID- 17112854 TI - Urinary albumin excretion in patients with diabetes after renal transplantation. AB - BACKGROUND: Many studies support microalbuminuria screening as the cornerstone of early detection of nephropathy in patients with diabetes mellitus (DM), but there are no studies that address its utility in the follow-up of renal transplant recipients with DM. MATERIALS AND METHODS: We retrospectively studied 104 subjects with DM who had undergone renal transplantation. Sixty three had a history of preexisting DM and 41 developed posttransplant diabetes (PTDM). We collected data on random urine albumin/creatinine (A/C) ratio, systolic blood pressure (SBP), serum creatinine concentration (Cr), and HBA1C. RESULTS: Mean subject age was 58 +/- 9.8 years; 63% received cadaveric grafts while 37% received living donor grafts. Mean follow-up was 7 years (range 1.6 to 15.6). Seventy percentage developed proteinuria over time; of these 62% developed microalbuminuria (A/C ratio 30 to 300 microg albumin/mg creatinine) and 38% developed macroalbuminuria (A/C ratio >300 microg/mg). Subjects with preexisting DM were as likely to develop an elevated A/C ratio as those with PTDM. Higher A/C ratios correlated with higher SBP (P < .01), with higher HBA1C (P < .036), and with higher Cr (P < .01). Lower A/C ratios correlated with more stable Cr over time (P < .01). CONCLUSIONS: Strict SBP and glycemic control are associated with a significantly lower A/C ratio in patients with DM after renal transplantation. Annual monitoring of A/C ratio in patients with DM after renal transplantation can identify candidates for stricter glycemic and blood pressure control, similar to current recommendations for all other patients with DM. PMID- 17112855 TI - Effects of pentoxifylline on the cytokines that may play a role in rejection and resistive index in renal transplant recipients. AB - Pentoxifylline (PTX) is a nonselective phosphodiesterase inhibitor that inhibits the production of TNFalpha and IL6 and IL-10 cytokines. In renal rejection TNFalpha, IL-6, and IL-10 may have important roles. In this study, 22 renal transplant recipients treated with tacrolimus, prednisolone, and mycophenolate mofetil were prescribed PTX (2 x 600 mg/d) for 3 months (GI), and 20 similar patients not receiving PTX were used as controls (GII). Stable subjects whose serum creatinine was lower than 1.8 mg/dL and were more than 6 months posttransplant, were enrolled into this study if the blood pressure was well controlled and there was no diabetes mellitus, infection, or inflammation. At the end of 3 months TNF-alpha decreased from 4.2 +/- 2.1 to 2.4 +/- 0.7 (P = .001) and 4.0 +/- 2.2 to 3.9 +/- 1.7 (P = .718), IL-10 also decreased from 3.90 +/- 1.9 to 2.38 +/- 0.6 (P = .001) and 4.02 +/- 1.6 to 3.82 +/- 1.5 (P = .225) in GI and GII, respectively. For IL-10 and TNF-alpha the alterations between baseline and the last visit of GI and GII were significant (P < .002 for all). Resistive index (RI) decreased in GI but the difference in alterations between baseline and the last visit of GI and GII was marginal. In summary IL-10 and TNF-alpha levels decreased in stable recipients treated with PTx. RI also decreased marginally secondary to PTx treatment. PTx was well tolerated and free side effects. PTx did not affect tacrolimus levels or other biochemical and hematological parameters. PMID- 17112856 TI - How to decrease cardiovascular mortality in renal transplant recipients. AB - Cardiovascular disease is the leading cause of death following renal transplantation, accounting for 40% to 55% of all deaths. An analysis in our center showed a 15% mortality in a cohort of renal transplant recipients followed for an average of 10 years. Various contributing risk factors of cardiovascular diseases in transplant recipients such as tobacco use, hypertension, hyperlipidemia, hereditary risk, diabetes, physical inactivity, obesity, dialysis duration, hyperuricemia, proteinuria, hyperhomocysteinemia, hyperparathyroidism, anemia; C-reactive protein level, and immunosuppressive regimen as well as some rare risk factors, such as cytomegalovirus infection, were evaluated in a population of 1200 kidney transplant recipients. Also we introduced methods for early detection, monitoring, and follow-up of proven risk factors of cardiovascular disease. PMID- 17112857 TI - Influence of methylprednisolone on plasma homocysteine levels in cadaveric renal transplant recipients. AB - The aim of the present study was to investigate plasma homocysteine levels in renal transplant recipients in the course of steroid-based or steroid-free immunosuppression. Data from 32 patients were retrospectively analyzed according to the steroid immunosuppressive regimen. The 20 recipients on methylprednisolone (MP) plus cyclosporine (CyA) or tacrolimus (TRL) (n = 20) showed similar creatinine levels when compared with those on calcineurin inhibitors plus mycophenolate mofetil (MMF; n = 12), (1.6 +/- 1.5 vs 1.6 +/- 0.4 mg/dL; P = NS) but significantly higher total plasma homocysteine (tHcy) levels (28.5 +/- 12.5 vs 16.3 +/- 5.5 micromol/L; P < .05). No differences of tHcy levels have been observed when patients were analyzed according to CyA- or TRL-based immunosuppression regardless of MP or MMF associations. Our data suggest that recipients, particularly those on steroid-based immunosuppression, should receive homocysteine-lowering treatment early after transplantation. PMID- 17112858 TI - Hepcidin, an acute-phase protein and a marker of inflammation in kidney transplant recipients with and without coronary artery disease. AB - BACKGROUND: In kidney transplant recipients, endothelial dysfunction and atherosclerosis are almost universal, as are cardiovascular complications. Inflammatory markers have been shown to play a role in the pathogenesis and progression of atherosclerosis, regarded as a chronic inflammatory condition. Iron metabolism is disturbed in chronic inflammatory diseases such as atherosclerosis. Hepcidin, the liver-expressed antimicrobial peptide, LEAP-1, is an acute-phase reactant produced in the liver that displays intrinsic antimicrobial activity. Cross-sectional study was performed to assess possible relations between hepcidin and inflammatory markers in kidney transplant recipients with versus without coronary artery disease (CAD). METHODS: Iron status, complete blood count, creatinine, albumin, and lipids were estimated using standard laboratory methods. Glomerular filtration rate (GFR) was calculated using the MDRD formula. Hepcidin, high-sensitivity C-reactive protein (CRP), IL-6, TNFalpha, and soluble receptor of transferrin were measured using commercially available kits. RESULTS: Kidney transplant recipients with CAD were older, and showed higher hepcidin, hsCRP, IL-6, TNFalpha, sTFR, ferritin, and lower cholesterol levels than did patients without CAD. Univariate analysis of values in kidney transplant recipients showed hepcidin to correlate significantly with total protein, ferritin, time after transplantation, creatinine, eGFR (simplified MDRD), cholesterol, neutrophil count, hsCRP, and IL-6. There were tendencies to correlate with TNFalpha. Multiple regression analysis showed that hepcidin was independently related to GFR, cholesterol, and hsCRP. CONCLUSIONS: Elevated hepcidin values in kidney allograft recipients may be due not only to impaired renal function, but also to a low-grade inflammatory state, as reflected by hepcidin correlations with hsCRP, IL-6, and ferritin. PMID- 17112859 TI - Simvastatin down regulates mRNA expression of RANTES and CCR5 in posttransplant renal recipients with hyperlipidemia. AB - Chemokines and hyperlipidemia are involved in the mechanism of chronic allograft nephropathy (CAN). In this study, the mRNA expression of RANTES and its receptor CCR5 on peripheral blood mononuclear cells were measured in renal transplant recipients with hyperlipidemia, and the effect of simvastatin treatment observed to investigate the mechanism and prevention of CAN. Sixty recipients selected from 167 renal transplant recipients were divided into two groups: group A without hyperlipidemia (n = 30) and group B with hyperlipidemia (n = 30). The control group consisted of 30 healthy volunteers. The recipients in group B were treated with simvastatin for 3 months. We estimated serum lipid levels and mRNA expressions of RANTES and CCR5. The mRNA expressions of RANTES and CCR5 were significantly higher in renal transplant recipients compared with controls. The expressions were much higher in group B than in group A patients. In group B patients, serum lipid levels decreased dramatically after simvastatin treatment. Meanwhile, the mRNA expressions of RANTES and CCR5 were reduced significantly after 1.5 months of simvastatin treatment to a level significantly lower than that in group A after 3 months of treatment. The increased expressions of RANTES and CCR5 mRNAs in renal transplant recipients with hyperlipidemia might be involved in CAN due to hyperlipidemia. Simvastatin seemed to reduce the chemokine transcripts in renal recipients with hyperlipidemia. PMID- 17112860 TI - Expression of transforming growth factor-beta receptor II mRNA in cyclosporine induced gingival overgrowth. AB - Gingival overgrowth (GO), characterized by increased cellular and extracellular matrix components in gingival tissue, is a frequent side effect of cyclosporine (CsA). In previous studies, elevated levels of transforming growth factor-beta (TGF-beta) have been detected in GO tissue, which led to the conclusion that TGF beta plays a major part in the pathogenesis. TGF-beta activity is mediated by three receptors; TGF-beta receptor II (TGF-beta RII), the most important, has been immunohistochemically detected in GO and normal gingival tissue. The aim of this study was to clarify whether TGF-beta RII is overexpressed in CsA-induced GO. The expression of TGF-beta RII mRNA in GO tissue of patients on CsA (n = 10, 5 women, aged 42.5 +/- 14.9 years) with renal transplantation (transplant duration 3.6 +/- 0.96 years) was compared with that in healthy gingiva of control subjects (n = 10, 5 women, aged 42.5 +/- 7.6 years). Semiquantitative reverse transcribed-polymerase chain reactions (RT-PCR) were applied with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as an internal standard. TGF-beta RII mRNA was readily detected in the GO tissue of patients on CsA. The level of TGF-beta RII mRNA relative to GAPDH in GO cases was not significantly higher than the relative TGF-beta mRNA level in normal gingiva (0.60 +/- 0.16 vs 0.52 +/- 0.19; P = .575). The precise mechanism of CsA-induced GO remains uncertain. According to our results, TGF-beta RII was not upregulated in CsA-induced GO, and may have no important role in this disorder. However, the involvement of TGF-beta in the molecular pathology of GO may be mediated via TGF-beta RI or RIII. PMID- 17112861 TI - Menstrual cycle and sex hormone profile in perimenopausal women after liver transplantation. AB - Excellent long-term outcomes of transplant patients let many female liver recipients experience perimenopausal problems. This study assessed menstrual patterns and sex hormone profiles in women of perimenopausal age who experienced end-stage liver failure treated by transplantation (OLT). MATERIALS AND METHODS: Menstrual patterns, sex hormone profiles, and biochemical parameters of liver function were analyzed before and after OLT in 13 liver-transplanted patients of perimenopausal age. Nineteen healthy perimenopausal women served as controls. RESULTS: The most common abnormality of the menstrual cycle observed in the study group was secondary amenorrhea, which affected six liver-transplanted women. Three months after OLT amenorrhea was still observed in six patients, regular menstrual cycles in six and irregular bleeding in one graft recipient. One year after transplantation regular menstruations were noted in four, irregular bleeding in four, and secondary amenorrhea in five liver-transplanted women. Similar levels of follicle stimulating hormone, luteinizing hormone, prolactin, progesterone and testosterone as well as lower levels of estradiol and DHEA sulfate were observed in patients with liver failure, both before and after grafting, compared with healthy women. After OLT E2 levels increased from 32.05 +/- 18.04 to 49.12 +/- 22.21. CONCLUSIONS: One year after OLT disturbances in menstrual patterns affect most (69%) perimenopausal female liver recipients. Both before and after OLT significantly lower levels of estradiol and DHEA-S were observed in transplanted patients compared with healthy controls. Hormonal therapy of amenorrhea or irregular menstruations may be required in that group of patients. PMID- 17112862 TI - Tamoxifen therapy in encapsulating sclerosing peritonitis in patients after kidney transplantation. AB - Sclerosing encapsulating peritonitis (SEP) is a serious complication of long-term continuous ambulatory peritoneal dialysis (CAPD) associated with obstructive symptoms and sclerosis of the peritoneal membrane. We present two cases that were successfully treated with tamoxifen and corticosteroids. Case 1: A 40-year-old patient developed end-stage renal failure (ESRF) and was managed with CAPD. He was hospitalized with symptoms of small bowel obstruction. He underwent laparotomy confirming the diagnosis of SEP. The patient was given tamoxifen 20 mg twice a day. Case 2: A 55-year-old patient with ESRF secondary to membranous glomerulonephritis. After having a cadaveric renal transplant in 1978 that failed 20 years later, the patient returned to CAPD. Six years later he had an uneventful kidney transplant and the peritoneal dialysis catheter was removed. However, 8 months later he presented with symptoms of small bowel obstruction and gross blood stained ascites. He also underwent a laparotomy that confirmed the diagnosis of SEP after biopsy. The patient was started on 20 mg of tamoxifen twice a day. Both patients' symptoms were improved gradually with an increase of serum albumin and body weight. Tamoxifen may be useful in the treatment of patients diagnosed with SEP. PMID- 17112863 TI - Renal artery aneurysm at the anastomosis after kidney transplantation. AB - Vascular complications represent serious problems after kidney transplantation. An aneurysm of the transplanted renal artery is an extremely rare but potentially devastating complication that which occurs in fewer than 1% of recipients. It can cause hypertension, functional impairment, and even graft loss. A 49-year-old man was admitted 6 months after his second renal transplantation. Duplex ultrasonography demonstrated an aneurysm at the anastomosis of the transplanted renal artery. The patient has not had any complaints. The function of the graft was stable. A computed tomography scan confirmed the diagnosis. Because of the high risk of rupture we decided upon surgical repair. During the operation, blood flow to the kidney was occluded; the graft was cooled with Euro-Collin's solution and ice-cold saline. After the resection there was enough usable arterial wall to construct a new anastomosis. The patient had an uneventful postoperative period, the serum creatinine decreased to the preoperative level, and the function of the graft was stable. Renal artery aneurysms represent high-risk complications. We decided on surgical repair, which was performed with simultaneous perfusion and cooling of the graft. There are only a few similar cases in the literature; it was the first operation using this method in our practice. Surgical reconstruction of a renal artery aneurysm, if feasible, is a safe procedure that prevents aneurysm rupture and saves the graft. PMID- 17112864 TI - Efficacy of interventional radiology procedures for the treatment of early ureteral complications after kidney transplantation. AB - Ureteral stricture and ureteral leakage are the most common early urological complications after kidney transplantation causing decreased urine output and increased serum creatinine and blood urea nitrogen. We report our experience with internal-external ureteral stent placement and ureteroplasty. MATERIALS AND METHODS: From August 1999 to January 2005, we treated nine patients presenting with stricture or leak. After an anterograde pyelogram, an internal-external nephrostomy catheter was inserted in all patients; in four patients we also performed ureteroplasty. RESULTS: The stricture and leak appeared from 12 to 93 days after kidney transplantation (mean = 39 +/- 29 days). After a mean of 80 +/- 43 days (range 25-141 days), the stent was successfully removed in seven patients (77%); no patient had a recurrence. The success was confirmed by a decline in serum creatinine (from 3.7 +/- 1.4 to 1.6 +/- 0.7 mg/dL) and blood urea nitrogen (from 54 +/- 24 to 28 +/- 7 mg/dL) with resolution of hydronephrosis on sonography. No procedure-related complications were observed. Surgical correction was necessary in two patients due to the persistence of a stricture. At long-term follow-up (50 +/- 17 months), seven kidneys were still functioning and two had failed due to chronic rejection. CONCLUSION: Nephrostomy catheter placement and ureteroplasty are safe, effective alternatives to surgery to treat early ureteral complications after kidney transplantation. Interventional radiology procedures reducing the morbidity and the likelihood of loss of graft function may improve graft and patient survival. PMID- 17112865 TI - Endoscopic management of ureteral complications following renal transplantation. AB - Management of ureteral complications after kidney transplantation can be done with a surgical, percutaneous, or endoscopic approach. The aim of this study was to determine the success rate of the endoscopic retrograde approach for the management of these complications following renal transplantation. We reviewed the records of 25 patients who underwent endoscopic management of ureteral complications after renal transplant between 1995 and 2005. Variables examined included timing of event following transplant, type of ureteral complication, equipment implemented in the procedure, operating time, success in stent placement, and complications. Initial approach was via rigid cystoscopy followed by flexible cystoscopy if needed. Initial attempts to intubate the ureteral orifice were by a flexible-tipped guide wire, and occasionally an angiocatheter guide was used for ultimate wire placement. Stents were positioned with fluoroscopic and direct visual guidance. Of 25 patients evaluated, five had a ureteral anastomotic leak with a mean time of presentation of 16.8 days. The remaining 20 patients suffered from ureteral obstruction revealed by hydronephrosis on a renal ultrasound prompted by a rising creatinine. Mean time of onset was 48 months. Although each was initially approached with rigid cystoscopy, 12 were converted to flexible cystoscopy for easier access to the ureteral orifice. Twenty of the 25 patients had successful stent placement with three failures in the ureteral obstruction group and two failures in the leakage group. Average operative time was 42 minutes. No intraoperative complications were experienced. Resolution of hydronephrosis in those with preoperative obstruction was noted and all stented urinary leaks resolved. PMID- 17112866 TI - Urine cytology as a useful screening method for polyoma virus nephropathy in renal transplant patients: a single-center experience. AB - Polyoma virus nephropathy occurs in 3% to 4% of renal transplant recipients, causing graft loss in 50% of cases. In this study we sought to identify the incidence of polyoma virus infection among our transplanted patients on the basis of age, sex, creatinine level, and postoperative period. During this study the 1086 urine samples collected from 362 patients were centrifuged and stained with the Papaniclaou method. All slides were classified as negative or positive (>1 decoy cell/sample). Among 1086 urine cytologies from 241 men and 121 women, decoy cells were identified in 26.6% (96) of patients, including 29.9% (n = 72) men and 20% (n = 24) women. The incidence of decoy cells (26.6%) was increased among men and associated with a longer transplantation period (P < .05). A significant relation was detected between older age and positive urine cytology. The patients with positive urine cytology for decoy cells showed a greater incidence of abnormal plasma creatinine values (26%) compared with patients showing a negative urine cytology (13.5%). In conclusion, identification of cells with viral inclusions (decoy cells) may help with the diagnosis of viral replication or active infection, therefore, routine urine cytology may be used as screening method for the detection of polyoma virus infection. PMID- 17112867 TI - Prevalence of human cytomegalovirus UL97 D605E mutation in transplant recipients in China. AB - Human cytomegalovirus (CMV) resistance to gancyclovir (GCV) occurs via mutation in the UL97 gene, ethylene diamine tetraacetic acid blood samples were obtained from 23 transplant recipients who received a GCV implant. A nested polymerase chain reaction amplifying UL97 codons 450 to 672 was performed. Nested amplifications were sequenced directly. No known UL97 GCV resistance mutations were found. Eighteen of 23 patients (78%) had revealed mutations at codon 605 (D to E). Mutant D605E may reverse to wild-type during the follow-up treatment. We conclude that human CMV UL97 D605E mutation occurred in Chinese transplant recipients. This mutation may be regarded as a natural sequence variant. PMID- 17112868 TI - Waiting for a liver transplant: psychosocial well-being, spirituality, and need for counselling. AB - The number of patients in need of a liver transplant vastly exceeds the number of available organs; the demand worldwide for organs leads to increased waiting times and mortality of patients on the waiting list. The aim of our study was to assess the psychosocial well-being of transplant candidates and their need for psychosocial counselling. METHODS: Sixty-nine liver transplant candidates were asked about their psychosocial well-being, quality of life, spirituality, and need for counselling assessed by interview and questionnaire (HADS-D, FLZ, LOT, SOC, SF-36, SBI-15R) during the initial evaluation procedure for organ transplantation as well as 3 and 6 months after listing. RESULTS: Candidates for a liver transplant exhibited a significant limitation in the levels of their quality of life and psychological well-being, compared with the community normal samples. They showed significantly higher levels of anxiety, but lower levels of spirituality. Almost half of the candidates (47%) expressed a need for counselling during the evaluation procedure. Patients with advanced diseases reported a lower need. Age and the personality-related "Sense of Coherence" correlated negatively with need for counselling. On the waiting list, psychosocial parameters and functions remained largely stable; the need for counselling decreased significantly. CONCLUSIONS: There is a relevant need for psychosocial counselling during the process of liver transplant evaluation. Need for counselling is associated with personality and age, as well as with somatic parameters. PMID- 17112869 TI - Efficacy of 6-month pretransplant abstinence for patients with alcoholic liver disease undergoing living donor liver transplantation. AB - PURPOSE: Questions have been raised regarding the ethics of liver transplantation in patients with alcoholic liver disease (ALD), including the fairness of cadaveric organ allocation to individuals who abuse alcohol and the efficacy of transplantation in these patients, many of whom may relapse. Living donor liver transplantation (LDLT) for ALD patients raises the similar ethical issues. ALD candidates for cadaveric liver transplants are required to abstain from alcohol for 6 months before being listed, but the efficacy of 6 months of abstinence in ALD patients receiving LDLT is not known. METHODS: We therefore determined the efficacy of 6 months of pretransplant abstinence in 15 ALD patients who underwent LDLT from February 1997 to December 2003. RESULTS: The Model for End-stage Liver Disease score was 24 +/- 10, and mean pretransplant abstinence period was 15 +/- 13 months, with 11 (73.3%) patients being abstinent for at least 6 months. Four patients received dual grafts, making the number of living donors 19: 12 children, two wives, one brother, three nephews, and one aunt. There were no unrelated donors. Three patients showed a relapse to alcohol drinking. The overall 1-, 3-, and 5-year survival rates were 100%, 100%, and 87.5%, respectively, and the cumulative 1-, 3-, and 5-year relapse rates were 6.7%, 20%, and 20%, respectively. The relapse rates in patients who did and did not maintain 6 months of abstinence were 9.1% and 50%, respectively; this difference was not significant (P = .154), likely due to the small sample size. Younger recipient age was a significant risk factor for alcohol relapse (40 +/- 8 years versus 53 +/- 6 years; P = .004). CONCLUSIONS: Pretransplant abstinence of 6 months seemed to be beneficial. For ethical reasons, a 6-month abstinence rule should be strictly observed in LDLT. PMID- 17112870 TI - Preoperative helical computerized tomography estimation of donor liver volume. AB - BACKGROUND: The purpose of this study was to evaluate the accuracy of spiral computed tomography (CT) and 3-D imaging models in measuring total and segmental liver volume in potential living donors. METHODS: A prospective study was undertaken to assess the correlation between the volumes of potential donor livers determined via helical CT and the actual volumes measured during operation in 150 donor candidates. Left-lateral segment (S2,3) or left-lobe (S2,3,4) transplantation was performed in 36 cases with 96 right-lobe liver transplants (S5,6,7,8). Ten donor candidates were refused owing to inadequate liver volumes, and 8 for other reasons. RESULTS: The regression analysis model showed a significant correlation between the preoperative CT estimates of graft volume and intraoperative weight measurement of harvested grafts in living liver donors (F: 5525.37; P < .05); 97.7% of changes in CT volume were explained by differences in graft mass (R2: 0.977). CONCLUSION: Preoperative estimation of segmental volumes of the donor liver is necessary to avoid donor-recipient size disparity, thereby preventing hepatic failure of donors after harvesting. It has a major impact on donor selection and type of surgical management. The accuracy of helical CT was high to determine total and segmental liver volumes. PMID- 17112871 TI - Impact of celiac axis stenosis on living donor hepatectomy. AB - BACKGROUND: Celiac axis stenosis was once cited as a reason for exclusion from living donor liver transplantation. Donor hepatectomy, however, leaves the pancreaticoduodenal artery arcade untouched, and theoretically, celiac axis stenosis has no impact on otherwise possible donors. METHODS: Among 350 consecutive adult living donors of liver transplantation at Tokyo University Hospital, we experienced 11 (3%) donors with celiac axis stenosis or occlusion due to the median arcuate ligament. RESULTS: Harvesting of the right liver was the most common procedure (n = 8), followed by harvesting of segments II and III (n = 2), and left liver (n = 1). The postoperative course was uneventful in all of the donors. CONCLUSIONS: Our results indicate that donor hepatectomy can be safely performed in the presence of significant celiac artery stenosis. PMID- 17112872 TI - Survival analysis of liver transplant patients in Canada 1997-2002. AB - Liver transplantation is an important health care issue for Canadians. Very few studies have assessed survival and determinants of survival in liver transplant patients in Canada. METHODS: We carried out an epidemiological analysis of 1 year survival and determinants of 1 year survival in liver transplant patients, using Canadian Organ Replacement Registry data (1997-2002). Survival curves were plotted by the Kaplan-Meier method. Cox proportional hazards analysis was applied to evaluate hazard ratios with different age groups, gender, ethnicity, blood groups, donor type, pretransplantation medical status, and HBV infection status. RESULTS: A total of 1164 liver transplant patients were included in the analysis. One-year survival rate was 84.7%. Male recipients had a 21% higher risk of developing organ failure than females. Recipients over 60 years of age had a 5% lower survival probability in comparison with recipients below 20 years of age. Pacific Islanders and Aboriginals had 32% and 9% lower survival probabilities, respectively, in comparison with Caucasians. Type B blood recipients had a 12% higher survival probability, whereas type AB blood recipients had a 7% lower survival probability compared with type O blood recipients. Twenty-six live organ recipients had 40% higher survival probabilities than 1138 cadaveric organ recipients. Patients with fulminant hepatitis (status 3F) had the highest survival, while patients with fulminant failure in ICU with intubation/ventilation (status 4F) had the lowest survival. One hundred sixty seven recipients with positive HBsAg antigen showed 10% lower survival probability than 997 cases with negative HBsAg antigen. CONCLUSION: In Canada, the first year survival rate is about 85%, which is comparable with other industrialized countries. Type of donor organs and recipient gender, ethnicity, ABO blood group, pretransplantation medical status, and HBV infection status had significant affects on the recipient survival. PMID- 17112873 TI - Results of biliary reconstructions in liver transplantation at our center. AB - Biliary complications are some of the most critical problems in liver transplantation. Despite various refinements in surgical technique, different types of liver transplantations are associated with significant numbers of biliary problems. In this study, we analyzed the results of biliary reconstructions in 127 liver transplant recipients at our center from April 2001 to May 2006. Through November 2004, we used different techniques for biliary reconstruction in 66 of these patients, including duct-to-duct (DD) anastomoses, Roux-en-Y hepaticojejunostomy (RYHJ), anastomoses over T tubes or stents, and anastomoses without stenting. During the first period, we used a DD anastomosis in 15 cadaveric whole liver grafts and in 25 right lobe and 12 left lobe or left lateral segment grafts from living-related donors. RYHJ was preferred in 2 cadaveric and 12 left lateral segment grafts. Beginning in November 2004, we employed intraoperative transhepatic biliary catheter insertion in 61 patients (29 children, 32 adults). In the most recent 61 cases of 13 liver grafts from cadavers and 48 from living-related donors, 14 patients (2 children and 12 adults) received whole-liver grafts, 22 (all adults) a right lobe, and 26 (all children) a left lateral or left lobe. Intraoperative transhepatic biliary catheter insertion was performed with DD anastomosis in 55 cases and with RYHJ in 6 cases. The mean complication rate decreased from 24% to 8.1% during the period using a new biliary reconstruction technique. Five biliary complications occurred in four patients. The new technique of biliary reconstruction using intraoperative biliary catheter insertion has significantly reduced the biliary complication rate. Transhepatic biliary stenting prevents biliary complications and maintains percutaneous access when problems arise. Intraoperative transhepatic biliary catheter insertion at the back table is a safe way to provide good biliary drainage after liver transplantation. PMID- 17112874 TI - Reappraisal of seventh-day syndrome following living donor liver transplantation. AB - Seventh-day syndrome (7DS) is characterized by sudden failure of a liver graft that had been working normally at about 1 week after transplantation, without an identifiable cause. A nonnegligible percentage of cadaveric liver transplants have shown this type of acute graft failure, whereas 7DS has not been reported after living donor liver transplantation (LDLT). Among 580 adult LDLT recipients in our institution between 1997 and 2003, 3 (0.5%) showed clinical sequences typical of 7DS. All three recipients showed similar but unique clinical sequences, consisting of initial uneventful recovery, dramatic rise of serum liver enzyme levels about 1 week later despite potent antirejection therapy, and subsequent graft loss. Liver biopsy findings were compatible with massive hemorrhagic necrosis. Sustained fever lasting for 2 days preceded deterioration of liver function. All three patients died prior to the opportunity for retransplantation. Our findings suggest that, as in cadaveric donor liver transplantation, 7DS can also occur following LDLT and that a preceding episode of sustained fever may be a prodrome of 7DS although its pathogenesis is yet poorly understood. PMID- 17112875 TI - Long-term outcomes in liver transplant patients with hepatic C infection receiving tacrolimus or cyclosporine. AB - Choice of calcineurin inhibitor may be a contributing factor to deteriorating patient and graft survival following liver transplantation for hepatitis C virus (HCV). In our multicenter, open-label LIS2T study, de novo liver transplant patients stratified by HCV status were randomized to cyclosporine or tacrolimus. Follow-up data were obtained in an observational study of 95 patients. Mean follow-up was 34 and 37 months, respectively, for cyclosporine-treated (n = 47) and tacrolimus-treated (n = 48) patients. In patients not receiving antiviral therapy, 22 of 31 given cyclosporine (72%) and 24 of 29 given tacrolimus (83%) had biochemical recurrence of HCV. In 68 patients with at least one biopsy, histological evidence of HCV-related hepatitis was present in 27 of 31 (87%) cyclosporine-treated patients and 37 of 37 (100%) tacrolimus-treated patients (P = .02, chi-square test). Three-year actuarial risk of fibrosis stage 2 was 66% with cyclosporine and 90% with tacrolimus; for fibrosis stage 3 or 4 it was 46% and 80%, respectively. Three graft losses were attributed to HCV recurrence in cyclosporine-treated patients and six in tacrolimus-treated patients. Tacrolimus may be associated with increased risk of histological HCV disease recurrence compared to cyclosporine. PMID- 17112876 TI - Domino liver transplantation with double piggyback: is this the best technique? A case report. AB - Sequential or domino liver transplantation is a well-established procedure for patients with familial amyloidotic polyneuropathy (FAP). Donation for domino liver transplantation imposed the resection of the inferior vena cava along with the liver, requiring complete suprarenal vena cava clamping and usually the use of venovenous bypass. We describe a successful case in which it was possible to perform the FAP hepatectomy by the piggyback technique. PMID- 17112877 TI - Suitable whole blood levels 2 hours after neoral in liver transplant patients: experiences at a single center. AB - Whole blood levels 2 hours after Neoral (C2) administration were observed to correlate better with area under the curve (AUC(0-4)) than trough levels (C0), suggesting that C2 may be the best single time point predictor of Neoral absorption. Owing to concerns about drug toxicity due to excessive immunosuppression, C2 adjustments to target blood levels may represent an advance. The present study measured C2 and levels to determine which correlated more closely with AUC(0-4). METHODS: Between August 2003 and July 2004, 40 adult liver transplantations were performed in our center. All patients received Neoral twice daily. They were maintained at a C0 level of about 200 ng/mL. C0 levels were measured daily. C2 levels were estimated on postoperative days 3, 5, 7, 14, and 28. AUC(0-4) performed on postoperative days 3, 7, and 28 was calculated using the trapezoidal rule. RESULTS: The mean AUC(0-4), C0, C1, C2, C3, and C4 were 1100.3 +/- 484.8 ng/mL, 197.1 +/- 84.7 ng/mL, 240.7 +/- 166.2 ng/mL, 307.8 +/- 162.6 ng/mL, 302.8 +/- 138.9 ng/mL, and 300.3 +/- 142.8 ng/mL, respectively. C2 correlated with AUC(0-4) (R2 = 0.868: P < .05) better than C0 (R2 = 0.245: P < .05), C1 (R2 = 0.604: P < .05), or C4 (R2 = 0.583: P < .05). CONCLUSIONS: Neoral dose monitoring according to a mean C2 range of 307.8 +/- 162.6 ng/mL correlated better with AUC(0-4). Further studies are required to determine suitable C2 levels in liver transplant patients. PMID- 17112878 TI - Postoperative severe pneumonia in adult liver transplant recipients. AB - Severe pneumonia in adult liver transplantation (OLT) recipients is a dangerous condition with significant morbidity and mortality. To analyze the risk factors for postoperative severe pneumonia in OLT patients, we collected data from 132 consecutive adult patients who underwent OLT between February 1999 and April 2004. According to the American Thoracic Society consensus statement, episodes of severe pneumonia were observed in 24 patients (18.2%). We retrospectively reviewed the etiology diagnosis, treatment, and prognosis of the 24 recipients. Bacteria were responsible for 95.8% of these episodes (23 of 24), fungi for 16.7% (4 of 24) and viruses for 4.4% (1 of 24). Twenty-six percent of the bacterial pneumonias were due to Streptococcus alpha hemolyticus. The mortality rate was 37.5% (9 of 24) for patients with severe pneumonia versus 7.4% (8 of 108, P = .004) for patients without pneumonia. Two cases with hepatorenal syndrome died, and three patients with coinfection by bacteria and fungi died. Acute rejection episodes occurred in 15 patients, four of whom died. Mechanical ventilation and tracheotomy were required in 13 cases (54%). Six who experienced prolonged intubation died. Sputum and pleural fluid cultures helped to establish a diagnosis in 91.7% (22 of 24) of cases. Twenty cases (83%) underwent reduction in the immunosuppressive regimen. Patient age, intraoperative transfusion requirements, extubation time, and hospital stay were fatal predictors of prognoses. We concluded that early detection of the responsible pathogen; timely and specific diagnosis; reduction in the immunosuppressive regimen; appropriate treatment with reliable, effective techniques; and implementation of sensitive culture-based antibiotics was an effective strategy to treat severe adult pneumonia in liver transplantation recipients. PMID- 17112879 TI - Pulmonary complications following adult liver transplantation. AB - PURPOSE: Pulmonary complications frequently occur after liver transplantation, but the risk factors associated with them have not been fully determined. We therefore sought to identify risk factors for pulmonary complications among adult liver transplant recipients. METHODS: We retrospectively reviewed the medical records of 128 consecutive adult patients who underwent 131 liver transplantations during 2001. We evaluated the incidence, time of onset, and outcome of radiographically determined pulmonary complications, as well as the factors predictive of infectious complications. RESULTS: Postoperative chest roentgenograms detected 68 cases of pulmonary complications, including pleural effusion (n = 50), atelectasis (n = 6), pneumonia (n = 6), pulmonary edema (n = 5), and acute respiratory distress syndrome associated with pneumonia (n = 1). Of the seven patients with pneumonia, five died. On univariate analysis the risk factors predictive for pneumonia were high serum creatinine and total bilirubin, hemodialysis at the time of occurrence, and history of acute rejection and on multivariate analysis increased total bilirubin and history of acute rejection. Pulmonary complications were dependent on the medical condition at the time of occurrence rather than on the preoperative condition. CONCLUSIONS: Although the incidence of pneumonia in liver recipients was relatively low, the mortality rate in patients who developed this complication was high. High-risk patients undergoing liver transplantation thus require early diagnosis and intensive treatment to diminish the morbidity and mortality associated with pulmonary complications. PMID- 17112880 TI - Factors related to post-liver transplantation acute renal failure. AB - Acute renal failure (ARF) after liver transplantation (OLT) is a common complication with severe impact on early and late prognosis of recipients. Factors predicting its incidence have not been fully identified due to the lack of a universal standard as well as the variance of data between transplant centers. To identify factors related to post-OLT ARF, we retrospectively collected materials on 89 patients, who underwent OLT from 1999 to 2001 in our center. Factors associated with post-OLT ARF were identified using univariate logistic regression. Significant factors were then entered into a multivariate logistic regression to identify factors independently associated with post-OLT ARF. Upon univariate analysis, intraoperative volume of blood transfusion (P = .041) and duration of operation (P = .005) were significant. ARF was associated with a poor prognosis (P < .001). Only duration of operation (P = .026) was an independent factor predicting the development of ARF. In conclusion, intraoperative volume of blood transfusion and duration of operation were factors contributing to post-OLT ARF in which the duration of the operation was an independent risk factor. The incidence of post-OLT ARF greatly increased recipient mortality in the early postoperative period. PMID- 17112881 TI - Study of the renal function in nonrenal organ transplantation. AB - Kidney disease after transplantation of a nonrenal organ has been described to be the result of the nephrotoxicity from the commonly used calcineurin-inhibitors as well as other factors. The aim of this study was to evaluate renal function and potential risk factors for the development of chronic renal failure among nonrenal organ recipients. We designed a single-center retrospective study including all 165 of our cardiac and liver recipients between February 1998 and October 2003, collecting clinical, analytic, and therapeutic data. We excluded double transplants and patients with survival less than 6 months. Creatinine clearance was calculated according to the Cockcroft-Gault and the Levey Modification of Diet in Renal Disease (MDRD)-5 equations. Although 165 patients received a cardiac or liver transplantation, 17 died in the first 6 months and three were double transplants; therefore we analyzed 145 patients: 107 (74%) cardiac transplantations and 38 (26%) liver transplantations. There were 106 male and 39 female recipients. The mean age (+/-SD) at the time of transplantation was 54 +/- 10 years and the mean follow-up was 2.9 +/- 1.7 years. Urinalysis before transplantation was only performed in 33 patients (22.8%) including three (2.1%) who had proteinuria. Serum creatinine increased until 12 months after transplantation (P < .001), then it recovered its average level. Creatinine clearance calculated using the aforementioned equations showed a similar pattern, with a progressive decline to 12 months (P < .05), with eventual stabilization or even improvement. The factors that we observed to increase the risk of renal damage were age, female sex, obesity, and the presence of proteinuria prior to transplantation. There was a good correlation (r = 0.96) between cyclosporine but not tacrolimus trough levels and serum creatinine at 48 hours after transplantation. PMID- 17112882 TI - New-onset diabetes mellitus after living donor liver transplantation: possible association with hepatitis C. AB - BACKGROUND: The relationship between hepatitis C virus (HCV) infection and new onset diabetes mellitus (NODM) after liver transplantation is a controversial issue. METHODS: A total of 223 adult living donor liver transplantation (LDLT) recipients followed for more than 6 months were analyzed for the prevalence of NODM. The prevalence was compared between 62 HCV-positive and 161 HCV-negative patients. All the HCV-positive patients underwent preemptive antiviral treatment with interferon alpha2b and ribavirin. RESULTS: Preoperative diabetes mellitus was more frequently observed in HCV-positive patients (18% vs 4%, P = .001). NODM occurred more frequently in HCV-positive patients (41% vs 22%, P = .003). Multivariate analysis, however, revealed that HCV was not a predictor for NODM. A comparison of 14 HCV-positive patients with persistent NODM and 48 patients without persistent NODM indicated that there was no significant difference in the frequency of the viral response to antiviral therapy nor in HCV-RNA levels. Impaired glucose tolerance did not impact postoperative survival after LDLT. CONCLUSIONS: HCV was not associated with the prevalence of NODM after LDLT. NODM did not influence patient survival. PMID- 17112883 TI - Is post-Lipiodol CT better than i.v. contrast CT scan for early detection of HCC? A single liver transplant center experience. AB - Hepatocellular carcinoma is a highly vascular neoplasm usually arising from a cirrhotic liver. Delayed, noncontrast, computed tomography (CT) imaging after 7 to 14 days reveals an oil-based contrast agent to be concentrated in the tumor but not in normal hepatic parenchyma. The aim of this study was to retrospectively correlate the post Lipiodol CT scan findings with respect to tumor size in the explanted liver. We retrospectively reviewed adult patients who had undergone orthotopic liver transplantation between November 1995 and December 2004 and also had an hepatic arteriogram with Lipiodol injection as part of their pretransplant workup. We calculated sensitivity, specificity, false-negativity, false-positivity, and accuracy of the test, as well as positive and negative predictive values. Lipiodol CT exam had sensitivity of 1.0; specificity of 0.6 with a calculated positive predictive value of 0.89 and a negative predictive value of 1.0. Overall accuracy of Lipiodol CT scan test was found to be 0.91, which was superior to an intravenous contrast CT alone. In conclusion, because of the higher sensitivity and accuracy values, hepatic arterial Lipiodol injection can be considered during the pretransplantation workup of high-risk cirrhotic patients, since the current model for End-stage Liver Disease scoring system for hepatocellular carcinoma is built on the ultimate bulk of the tumor. Further multicenter, controlled, large-volume prospective studies are warranted to verify this observation. PMID- 17112884 TI - Myocardial mechanisms causing heart failure early after cardiac transplantation. AB - Early after heart transplantation, some patients have heart failure (HF) with preserved left ventricular ejection fraction (LVEF), in the absence of rejection. The purpose of this study was to define the mechanisms causing HF early after transplantation and to determine whether these mechanisms involve changes that occur in active or passive myocardial properties. Eleven consecutive patients 1 week after heart transplantation underwent right heart catheterization and echocardiography with an endomyocardial biopsy. Hemodynamic measurements were obtained at spontaneous heart rate, and then were repeated at three atrially paced rates increased in 20-bpm increments above spontaneous heart rate. At baseline, 5 patients (group 1) had clinical HF and a pulmonary capillary wedge pressure (PCWP) > or = 16 mmHg, and 6 patients (group 2) had no clinical evidence of HF and a PCWP < 16 mmHg. LVEF was normal in all 11 patients. The relationships between cardiac index versus heart rate (HR) and PCWP versus HR were normal in all 11 patients. These normal function-versus-frequency relationships suggested that there were no significant abnormalities in the active myocardial processes of contraction or relaxation. In group 1 patients, the PCWP was significantly increased but the left ventricular end diastolic dimension was normal, suggestive of diastolic stiffness. Early after transplantation, there was a significant increase in LV wall thickness in group 1 patients as compared with preexplantation values despite myocardial biopsies in all 11 patients, showing no evidence of rejection, cardiomyocyte hypertrophy, or interstitial fibrosis thus suggestive of myocardial edema. PMID- 17112885 TI - Extracorporeal membrane oxygenator assistance as "bridge" to combined heart and liver transplantation. AB - Heart transplantation is accepted worldwide as the treatment of choice for patients with end-stage heart disease who have a life expectancy of less than 1 year. Mechanical Cardiac assistance might be necessary as a bridge to transplantation and, in selected cases, to multiorgan transplantation. We report a successful case of a 15-day extracorporeal membrane oxygenator assistance as bridge to combined heart and liver transplantation in a young man with dilated cardiomyopathy and chronic hepatitis C virus infection. PMID- 17112886 TI - Initial single-center experience with sirolimus after lung transplantation. AB - BACKGROUND: Standard immunosuppression after lung transplantation includes calcineurin inhibitors, mycophenolate mofetil, and steroids. Long-term survivors of lung transplantation are often confronted with chronic kidney disease, by definition related to the intake of calcineurin inhibitors. Sirolimus has been increasingly proposed as an alternative immunosuppressive agent due to its absence of nephrotoxicity, which could be used in selected patients. METHODS: We prospectively administered sirolimus as an alternative to calcineurin inhibitors in 10 lung transplantation recipients with persistent drug nephrotoxicity. They were switched from tacrolimus to sirolimus. Four patients also had bronchiolitis obliterans syndrome. The conversion scheme consisted of an immediate stop of tacrolimus and an 6 to 8-mg loading dose of sirolimus, followed by 4 mg/d. After 5 days, the sirolimus dose was adjusted to maintain trough levels between 12 and 18 ng/mL or 6 and 12 ng/mL for combined sirolimus and tacrolimus. Patients were monitored for renal and graft function as well as clinical status. RESULTS: A significant decrease in creatinine was observed after 1 week of treatment (P = .011). Azotemia decreased after 1 month, remaining stable (P < .01). Pulmonary function tests did not show significant modification from before sirolimus, inception in patients with or without bronchiolitis obliterans syndrome. There were seven infections. One patient died of complications related to bronchiolitis obliterans. CONCLUSION: Sirolimus was a useful alternative immunosuppressant, allowing significant tacrolimus withdrawal in transplant recipients with renal impairment. Sirolimus administration allowed recovery of renal function with low morbidity; it was useful for rescue of chronic renal impairment after lung transplantation. PMID- 17112887 TI - Elevated levels of serum interleukin-6 are associated with low grade cellular rejection in patients with heart transplantation. AB - Endomyocardial biopsy is the gold-standard procedure to diagnose acute cellular rejection after heart transplantation. This study assessed whether the blood levels of cytokines involved in inflammation and immune activation are useful to detect the presence of acute cellular rejection. METHODS: Blood specimens collected before 275 endomyocardial biopsies in 66 patients were assayed for levels of TNFalpha, IL6, IL1beta, and IL2 receptor. The biopsies were grouped according to the presence (n = 41) or absence (n = 234) of acute cellular rejection grade > or = 3A of the International Society for Heart and Lung Transplantation. We compared the levels of cytokines in the two groups. RESULTS: Circulating IL6 levels were significantly higher when there was a low grade (0-2) cellular rejection in the biopsy versus the group of biopsies grade > or = 3A (19.8 +/- 27 versus 12.9 +/- 10 pg/mL; P = .001). An IL6 level higher than 30 pg/mL showed a negative predictive value of 95% for the presence of acute rejection grade > or = 3A. CONCLUSION: In heart transplant patients, high levels of serum IL6 were associated with low grade cellular rejection. Determination of IL6 levels may be useful to reduce the number of endomyocardial biopsies during follow-up in these patients. PMID- 17112888 TI - Low levels of high-density lipoprotein cholesterol are associated with vascular remodeling in cardiac transplant recipients. AB - BACKGROUND: Early atherosclerosis may be associated with compensatory vessel enlargement, termed positive remodeling. Enlarged brachial artery diameter has been reported in patients with risk factors for atherosclerosis and in individuals with coronary atherosclerosis, indicating that brachial artery enlargement is a marker for the presence of atherosclerotic changes. Cardiac transplant recipients often have abnormal lipid levels, but the effect of specific lipid abnormalities on vascular remodeling in this population has not been evaluated. This study examined the relationship between lipid levels and brachial artery diameter in cardiac transplant recipients. METHODS: Thirty-five stable cardiac transplant recipients underwent high-resolution brachial artery ultrasound to evaluate resting brachial artery diameter. Levels of high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and triglycerides were determined and the presence of other cardiac risk factors was assessed. RESULTS: Brachial artery diameter was larger (4.3 +/- 0.1 mm) in subjects with low levels of HDL-C (< 40 mg/dL, n = 11) compared to subjects with high HDL-C (> or = 40 mg/dL, n = 24), who had a mean brachial artery diameter of 3.7 +/- 0.1 mm (P = .006). Neither high LDL-C (> or = 100 mg/dL) nor high triglycerides (> or = 200 mg/dL) were associated with differences in brachial artery diameter. Multivariate analysis demonstrated that the relationship between low HDL-C and increased brachial artery diameter was independent of body surface area or statin use. CONCLUSIONS: Low levels of HDL-C are an independent predictor of brachial artery enlargement in stable cardiac transplant recipients. These findings suggest that suboptimal HDL-C levels may be associated with the development of vascular remodeling and atherosclerosis in this population. PMID- 17112889 TI - Neutrophil oxidative metabolism in diabetic patients undergoing pancreas transplantation. AB - A pancreas transplantation is the only therapy capable of returning a constant, physiological euglycemic state to diabetic patients. Considering the clinical controversies in the study of infection in diabetes and the recognized effect of insulin on the oxidative metabolism of glucose in phagocytes, the present study sought to evaluate the formation of intraphagocytic oxygen-free radicals in diabetic patients undergoing simultaneous pancreas kidney transplantation (SPK). METHODS: Twenty-five diabetic patients undergoing SPK were compared with 25 normal individuals. Evaluation of the oxidative metabolism of leukocytes was performed using the NBT test. RESULTS: The abnormality in the pretransplant counts (19.32%-28.2%) reached normal levels at 48 hours after transplantation (45.11%-76.25%) and was maintained to the 5th day (46.28%-76.20%). CONCLUSION: An SPK in a diabetic patient normalized the formation of intraphagocytic oxygen-free radicals. PMID- 17112890 TI - Cryopreservation of insulin-producing cells microencapsulated in sodium cellulose sulfate. AB - INTRODUCTION: Diabetes mellitus may be treated with pancreatic islet cell transplantation. The use of xenogenic islet cells may overcome the shortage of human donor organs. Microencapsulation seems to be a promising method for immunoprotection. Since isolation, purification, encapsulation, and transplantation of islet cells are labor-intensive, cryopreservation has emerged as an attractive system for islet banking. In this study sodium cellulose sulfate (NaCS), a novel method for microencapsulation of islet cells, was tested for its capability to protect cells during cryopreservation. METHODS: HIT-T15 cells were microencapsulated in NaCS. Cells were frozen and thawed using three different media containing varying amounts of dimethylsulfoxide (DMSO) and glycerol. Cell viability and cell growth were monitored using 3-(-4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide before freezing and 1 week after thawing. RESULTS: NaCS did not show any negative impact on the growth rates of encapsulated HIT-T15 cells compared with nonencapsulated controls. Nonencapsulated cells were adequately cryopreserved by both DMSO- and glycerol-containing freezing media. DMSO was not suitable for cryopreservation of encapsulated HIT-T15 cells, whereas glycerol seemed to produce no considerable cell loss during freezing and thawing. DISCUSSION: Islet banking of cells encapsulated in NaCS was feasible. Microencapsulation did not harm islet cell recovery. As NaCS is less immunogenic and more biocompatible than other materials used for microencapsulation, it may be a promising method for immunoisolation of islet cells to replace the endocrine pancreas in a physiological way. PMID- 17112891 TI - Pancreatic islet cells antibodies in diabetic patients submitted to pancreas transplantation. AB - Several pieces of evidence suggest an autoimmune etiology of diabetes mellitus type 1. To trace patients who are susceptible to the disease, we utilized islet cells antibodies (ICAs). The aim of this study was to evaluate the presence of ICAs among diabetic patients undergoing simultaneous transplantation of the pancreas and kidney (SPK). Twenty-six diabetic patients received an SPK, 12 of whom were included in this analysis. The indirect immunofluorescence method was utilized for quantitation of ICAs. The types of ICAs were no different following transplantation of the pancreas. The serum levels of pre-existent ICAS in diabetic patients undergoing SPK with immunosuppression were not reduced, and they did not interfere with the function of the implanted pancreas over a period of 60 days. PMID- 17112892 TI - Assessment of 18F-FDG-leukocyte imaging to monitor rejection after pancreatic islet transplantation. AB - AIM: We sought to investigate the feasibility of 18F-FDG-leukocyte imaging to detect islet rejection. METHODS: Two thousand Sprague-Dawley (SD, syngeneic group) or Lewis (allogeneic group) islet equivalents were intraportally injected into SD rat recipients. Four and 7 days after transplantation, 10(8) 18F-FDG labeled splenocytes were injected into the jugular vein. Splenocytes were harvested from naive or sensitized (12 days after intraportal transplantation of 2000 Lewis IEQ) SD rats. Positron emission tomography (PET) imaging was started 5 minutes after splenocyte infusion and performed hourly for 4 hours. RESULTS: One hour after splenocyte injection, FDG was mainly detected in the heart and lungs. It was then further distributed to other organs, and from the second hour, the highest tracer concentration was located in the abdomen. Liver FDG uptake was similar between syngeneic, allogeneic, and sensitized allogeneic groups at 4 and 7 days after islet transplantation. DISCUSSION: No islet rejection was detected by 18F-FDG-leukocyte imaging. The amount of transplanted tissue was only few millilitres and the additional related inflammation in case of rejection is small and difficult to detect. The liver showed a relatively high spontaneous tracer uptake; the related background prevented detection of a potential increase in tracer uptake in cases of islet rejection. PMID- 17112893 TI - Curcumin inhibits in vitro MCP-1 release from mouse pancreatic islets. AB - OBJECTIVES: Monocyte chemoattractant proteins (MCP-1) belongs to the CC family of chemokines secreted from islets of the pancreas, producing recruitment of inflammatory cells leading to an acute immune response with graft rejection in clinical transplantation. Expression and release of many inflammatory cytokines and chemokines, including MCP-1 is regulated by the nuclear factor (NF)-kappaB pathway. Curcumin is an NF-kappaB inhibitor with a variety of biological activities anti-inflammatory, antitumor, antioxidant, and antichemotactic effects. The aim of this study was to examine the effect of curcumin on in vitro MCP-1 release from pancreatic islets. METHODS: Mouse pancreatic islets in 18-hour cultures were treated with 0 or 10 or 20 micromol/L curcumin and with LPS for an additional 24 hours. MCP-1 levels in culture supernates of islets with versus without curcumin treatment were measured by an ELISA assay. RESULTS: We observed that curcumin at the concentration of 20 micromol/L significantly decreased MCP-1 release from mouse islets compared to the control group (P = .005). In addition at both of 10 micromol/L and 20 micromol/L curcumin concentrations there was a decreased level of MCP-1 released from LPS-treated versus control islets (P = .01). PMID- 17112894 TI - Validation of the scoring system for standardization of the pancreatic donor for islet isolation as used in a new islet isolation center. AB - BACKGROUND: The aim of this study was to evaluate the effectiveness of the Edmonton Donor Scoring System for use in our much less active islet center. Because the ability to recognize an appropriate donor may help to achieve consistent and predictable success of pancreatic islet isolation, it should lead to increased effectiveness and lower cost. MATERIAL AND METHODS: Charts of 36 consecutive pancreas donors were reviewed to assess the donor points (DP). DP ranged from 0 to 100 based on donor age, body mass index, cause of death, social and medical history, hospital stay, vasopressor dosages, laboratory tests, cold ischemia time and procurement team, as well as pancreas size, consistency, fat content, damage, and quality of procurement and packing. RESULTS: Successful isolation was achieved in 39% of donors (14 of 36), a value similar to that achieved in Edmonton (40%). We used the optimal cutoff value (DP = 79) proposed by the Edmonton group. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 66%, 75%, 57%, 82% and 72%, respectively. Successful islet isolation from poor or marginal donors (DP < 49.5 and 50 to 59.5) was 0% and 28.6% respectively; it was 63% and 100% in optimal donors (DP = 80 to 89.5 and 90 to 100). We concluded that islet isolation success correlated with the previously proposed donor scoring system. CONCLUSIONS: The Donor Scoring System can be successfully implemented regardless of the level of activity of an experienced isolation center. This system permits identification of a suitable donor prior to organ processing. It may guide a center's donor selection strategy based on its goals and its budget. PMID- 17112895 TI - Long-term survival of cardiac allografts induced by cyclophosphamide combined with CTLA4Ig-gene transfer mediated by adenoviral vector. AB - There is a need to achieve donor-specific tolerance in clinical organ transplantation, where potential benefits remain overshadowed by chronic rejection and the side-effects of long-term immunosuppressive therapy. It is known that the mature immune system in mice can be reprogrammed to accept a foreign graft as if it was "self". The AdCTLA4Ig-mediated gene transfer (SC) + cyclophosphamide (CP) treatment alone prolongs allograft survival but does not induce tolerance. However, in our study, the AdCTLA4Ig-mediated gene transfer combined with SC + CP treatment yielded significantly prolonged mean survival times (149.7 +/- 18.0 days), while those in the untreated or AdLacZ treated mice were rejected in normal fashion (5.3 +/- 0.5 and 5.2 +/- 0.4 days, respectively), and survival in the AdCTLA4Ig or SC + CP treated groups were 45.7 +/- 9.6 or 50.2 +/- 5.3 days, respectively. In conclusion, a protocol of AdCTLA4Ig + SC + CP improved the survival of DA-->LEW cardiac allografts. PMID- 17112896 TI - Administration of donor-derived mesenchymal stem cells can prolong the survival of rat cardiac allograft. AB - BACKGROUND: Mesenchymal stem cells (MSCs) are multipotent adult elements that have recently been shown to have profound immunomodulatory effects both in vitro and in vivo. Herein we have examined the impact of intravenous infusion of donor MSCs on the survival of transplanted hearts in a rat allograft model. METHODS: Recipient Fisher344 rats were transplanted with hearts from inbred Wistar rats. Wistar rat MSCs were infused via the tail vein at designated intervals. In vitro mixed lymphocyte reaction (MLR) and cell-mediated lympholysis (CML) assays were performed to assess whether MSCs downregulated T-cell responses in vivo. Real time polymerase chain reaction (PCR) was used to analyze the Th1/Th2 balance in MSC-treated and control groups. RESULTS: The MSCs cultured in vitro exhibited multipotential for differentiation. Survival of the allografts was markedly prolonged by administration of MSCs compared with the controls, namely mean survivals of 12.4 vs 6.4 days, respectively. Real-time PCR showed a shift in the Th1/Th2 balance toward Th2. By MLR and CML assays, untreated control rats showed greater alloreactivity than did MSC-treated rats. CONCLUSION: Our results indicated that MSCs suppressed allogeneic T-cell responses both in vitro and in vivo. Intravenous administration of MSCs prolonged the survival of transplanted hearts, possibly by induction of allograft tolerance through changing the Th1/Th2 balance. PMID- 17112897 TI - The suppressive effect of resveratrol on protein kinase C theta in peripheral blood T lymphocytes in a rat liver transplantation model. AB - Our aim was to study the effect of resveratrol on the expressions of protein kinase C isotypes (PKC alpha, theta) in peripheral blood lymphocytes and on the expression of IkappaB kinase-beta (IKK beta) in lymphocytes in allografts in a rat liver transplantation model. METHODS: Orthotopic liver transplantations (OLT) were performed from Sprague Dawley rats to Wistar rats. The recipients were divided into two groups after OLT. In the RES group, resveratrol was given intraperitoneally once a day (100 mg kg(-1)) after OLT, whereas in the control group vehicle buffer was given. The expressions of PKC alpha, theta in peripheral blood lymphocytes, expression of IKK beta in lymphocytes in allograft, and survival periods were compared between the groups. RESULTS: The mean survival period after OLT in the RES group was significantly longer than that in control group (P < .05). On posttransplant day 7, the expression of PKC theta in peripheral blood lymphocytes in the RES group was significantly decreased compared with that in the control group (P < .05), whereas there was no obvious difference in the expressions of PKC alpha between the two groups (P > .05), and the positive rate of IKK beta protein in lymphocytes in allografts in RES group was significantly decreased compared with that in the control group (P < .05). CONCLUSION: Resveratrol showed an immunosuppressive effect on lymphocytes for allograft rejection in the rat. Down-regulation of the expression of PKC theta in peripheral blood lymphocytes may be part of the mechanism. PMID- 17112898 TI - The study of peripheral blood mononuclear cell MHC I and MHC II gene mRNA expression in acute graft rejection. AB - BACKGROUND: Early diagnosis of acute graft rejection is important in the clinic. To explore a reliable diagnostic marker, we selected skin-grafted rabbits as an animal model to study peripheral blood mononuclear cell (PBMC) major histocompatibility complex 1 (MHC I) and MHC II gene mRNA in acute graft rejection (AGR). METHODS: Fifteen New Zealand white rabbits were randomly divided into three groups to observe skin graft rejection: three rabbits were in the autograft control group; six rabbits in a cyclosporine (CsA) treated allografted group; and the other six rabbits in untreated allografted group. The CsA-treated allografted group was given CsA (5 mg/kg) daily intramuscularly. PBMC samples were obtained every 2 days to detect by real-time polymerase chain reaction, PBMC MHC I and MHC II gene mRNA. RESULTS: MHC I and MHC II gene mRNA levels did not show any obvious change in the autografted controls. MHC I gene mRNA levels showed a slow increase in the CsA-treated allografted group, but no obvious change in the untreated allografted group. MHC II gene mRNA reached the highest level at 2 to 3 days before graft rejection appeared macroscopically in the CsA treated allografted group and untreated allografted group, then decreasing to a low level. CONCLUSION: Compared with MHC I gene mRNA expression, PBMC MHC II gene mRNA expression may be considered to be an earlier marker for AGR. PMID- 17112899 TI - Effect of FTY720 in rat small bowel transplantation: apoptosis of crypt cells and lymphocytes in gut-associated lymphoid tissues. AB - AIM: We investigated the extent of apoptosis in crypt cells and Peyer's patches (PPs) during small bowel allograft rejection in rats to examine the effect of FTY720 during rejection. METHODS: Orthotopic small bowel transplantations (SBTs) were performed from BN to LEW rats. Isografted animals served as controls. Three groups of SBT animals were studied on days 3, 5, and 7 after operation: isograft, untreated allograft, allograft with FTY720. FTY720 was orally administered by gavage (1 mg/kg/d) to allograft recipients on 7 consecutive days. Cryostat sections were prepared from grafts, including PPs. An in situ end-labeling (ISEL) technique was used to detect apoptotic cells. Indirect immunoperoxidase staining was also performed using monoclonal antibodies against rat Fas/Fas-L. RESULTS: Graft survival was prolonged in the FTY720-treated group. The number of ISEL positive enterocytes in the allografts increased significantly on days 3, 5, and 7 compared with the isograft group. In the FTY720-treated group, the number of ISEL-positive enterocytes in the allografts was down-regulated significantly on days 3, 5, and 7 compared with untreated allograft group. In the PPs, the number of ISEL-positive mononuclear cells increased significantly in the allografts compared with the isograft group. In the FTY720-treated groups, the number of ISEL-positive mononuclear cells were down-regulated significantly in the allografts compared with the untreated allograft group. The number of Fas/FasL positive enterocytes were increased significantly in allografts compared with isograft group. In FTY720-treated groups, the number of Fas/FasL-positive enterocytes were down-regulated significantly on day 7 compared with the untreated allograft group. In the PPs, Fas/FasL-positive mononuclear cells also increased significantly on day 7 in the allografts compared with isografts. In the FTY720-treated groups, Fas/FasL-positive mononuclear cells were down regulated significantly in the allografts compared with the untreated allograft group. CONCLUSIONS: The number of apoptotic enterocytes, lymphocytes, and Fas/FasL-positive lymphocytes increased during small bowel graft rejection. FTY720 prevented up-regulation of the number of apoptotic enterocytes, lymphocytes, and Fas/FasL-positive lymphocytes while also prolonging small bowel allograft survival. PMID- 17112900 TI - Intrathecal implants of microencapsulated xenogenic chromaffin cells provide a long-term source of analgesic substances. AB - Adrenal medullary chromaffin cells secrete several neuroactive substances including catecholamines and opioid peptides that produce analgesic effects in the central nervous system. This study was designed to investigate whether intrathecal microencapsulated chromaffin cells could release analgesic materials producing antiallodynic effects on the chronic neuropathic pain in rats induced by chronic constriction injury (CCI) of the sciatic nerve. Prior to intrathecal implantation, chromaffin cells were encapsulated with alginate and poly-L-lysine to protect them from the host immune system. Behavior tests were performed before CCI, 1 week later, and at 4, 7, 14, 21, 28 days postimplantation. At the end of study, we performed cerebrospinal fluid (CSF) collection and implant retrieval. We observed that intrathecal implantation of encapsulated xenogenic chromaffin cells reduced the mechanical and cold allodynia in a model of neuropathic pain. CSF levels of catecholamines and metenkephalin in the rats that received implants were higher than the controls. In addition, we observed chronic survival of implants. These results suggested that intrathecal microencapsulated chromaffin cells may represent a new approach to chronic neuropathic pain management. PMID- 17112901 TI - Molecular characterization of the porcine endogenous retrovirus subclass A and B envelope gene from pigs. AB - Xenotransplantation of porcine organs has the potential to overcome the current critical shortage of allogenic organs for transplantation in humans. However, the existence of porcine endogenous retroviruses (PERVs) presents a problem for the clinical use of xenografts from pigs. In an attempt to understand the molecular characteristics of PERVs, we cloned the PERV env gene from six pig breeds (ie, Berkshire, Duroc, Landrace, Yorkshire, and two types of miniature pigs) in Korea. A total of 141 env clones were isolated and their sequences were analyzed. Phylogenetic analyses of these genes revealed the presence of PERVs, from both classes A and B, in 54% and 46% of the env clones, respectively. Among these clones, 37 isolates had the correct open reading frame (ORF; 27 clones in subclass A and 10 clones in subclass B), while the others had premature termination. These PERV nucleotide sequences can be used in a database for comparisons of PERV distribution among different pig breeds and for monitoring PERV infection using isolates with functional ORFs. Recombinant envelope of subclass A and B with functional ORF was expressed by vaccinia virus systems. Additionally isolated env clones can be used for various experiments, such as PERV control and infectivity tests, and may enhance the understanding of molecular mechanisms through pseudotyped PERV viruses. PMID- 17112902 TI - Novel magnetic rings for rapid vascular reconstruction in canine liver transplantation model. AB - OBJECTIVE: Magnetic rings were used for rapid vascular reconstruction in a canine liver transplantation model. MATERIALS AND METHODS: Thirty-two adult mongrel dogs weighing 13 to 16 kg were randomly selected as donors or recipients of transplantations. The recipients were randomly divided into two groups: group A (n = 10) had magnetic rings used for vascular reconstruction without venovenous bypass; group B (n = 6) had vascular reconstruction performed by continuous suturing with splenojugular venovenous bypass. RESULTS: In group A, the entire operative period was 3.24 +/- 0.49 hours, the durations of clamping the portal vein and the infrahepatic vena cava of the recipient were 5.89 +/- 2.27 minutes and 3.89 +/- 0.73 minutes, respectively. In group B, the entire operative period was 4.12 +/- 0.51 hours with the duration of clamping portal vein and infrahepatic vena cava, 28.33 +/- 6.04 minutes and 12.16 +/- 3.72 minutes (P < .01 vs group A). In group A, mean arterial pressure dropped during the anhepatic phase but recovered quickly after reperfusion. The fluid infusion was about 730.56 +/- 50.56 mL in the group A and a pressor agent was unnecessary. In group B, blood pressure dropped during the anhepatic phase and slowly recovered. The fluid infusion was about 2241.67 +/- 390.78 mL and a pressor agent was used to maintain the blood pressure of the recipient. No twist or thrombus was discovered in the anastomoses group A and the endothelium at the site of anastomosis was entire. In group B, errhysis was common in the anastomotic stomas. Nine of 10 dogs in group A survived more than 3 days, the longest being 8 days, whereas four of the six dogs in group B survived less than 3 days. CONCLUSION: The results showed that the anhepatic time was significantly shortened (about 5.89 +/- 2.27 minutes) in group A compared with group B and venovenous bypass was unnecessary. Magnetic rings could be used for rapid vascular reconstruction in canine liver transplantation model. The long-term results of this procedure should be clarified before it is applied in clinical practice in the future. PMID- 17112903 TI - Heterogeneity of damage between segments of rat liver after inflow-outflow obstruction. AB - BACKGROUND: Total vascular exclusion (TVE) causes warm liver ischemia. The complete explanation of the events during inflow and outflow obstruction of the liver during selective TVE has not yet been studied. The aim of this study was to investigate the liver injury caused by inflow-outflow obstruction in the rat liver. MATERIALS AND METHODS: Forty Wistar-Albino rats were divided into four groups. Liver inflow occlusion (groups A and C) or inflow-outflow occlusion (groups B and D) was applied for 30 minutes. Samples were collected at the end of the ischemia period. We examined oxidative injury in the liver tissue and liver histopathology. RESULTS: Oxidative stress and histopathologic alterations were more prominent with TVE application. Significant alterations were shown in hepatic superoxide dismutase, glutathione, and glutathione S-transferase levels. Central segments of the rat liver were affected significantly from inflow occlusion, whereas dome segments were significantly damaged from inflow-outflow occlusion. CONCLUSIONS: Inflow-outflow occlusion of the liver caused more tissue damage compared with inflow occlusion. The pattern of distribution of the damage due to TVE seemed different from other well-known ischemia-reperfusion injuries. PMID- 17112904 TI - Differentiation of hematopoietic stem cells into hepatocytes in liver fibrosis in rats. AB - It has been reported that hematopoietic stem cells (HSC) can differentiate into hepatocytes in the normal liver and in some pathologic environments. The aim of this study was to investigate whether HSC can differentiate into hepatocytes in cases of established liver fibrosis. Rat liver fibrosis was induced by subcutaneous injection of tetrachloride (CCl4). Thy+ CD3- CD45RA- HSC in bone marrow cells, which had been enriched by fluorescence-activated cell sorting (FACS), were labeled with PKH26-GL, and autologously transplanted into CCl4 treated rats. The expressions of albumin (Alb), cytokeratin 8 (CK8), and alpha smooth muscle actin (SMA) were determined by immunofluorescence methods. The PKH26-GL labeled Thy+ CD3- CD45RA- HSC expressed the hepatocyte-specific markers Alb and CK8, but did not express alpha-SMA in liver fibrosis. Thy+ CD3- CD45RA- HSC differentiated into hepatocytes, but not into hepatic stellate cells. In conclusion, autologous stem cell transplantation may be helpful to treat hepatic fibrosis. PMID- 17112905 TI - Evaluation of wound healing effect on skin-defect nude mice by using human dermis derived mesenchymal stem cells. AB - We report that human dermis-derived mesenchymal stem cells (hDMSCs) possess differentiation potential of epidermis facilitating wound healing in skin-defect nude mice in combination with the treatment using gelatin/thermosensitive poly N isopropylacrylamide (pNIPAAm)/polypropylene (PP). The results showed that the rate of cell growth and wound recovery in the hDMSC and gelatin/pNIPAAm/PP treated group was significantly greater than those in the gelatin/pNIPAAm/PP treated only group (P < .01). The reepithelialization marker of human pan cytokeratin was also significantly increased on days 14 and day 21 in the wound site of hDMSCs and gelatin/pNIPAAm/PP-treated group. Furthermore, the stem cell marker of human CD13 gradually decreased during the period of wound healing. In sum, this novel method provided a transferring system for stem cell therapy, maintaining its temperature-sensitive property of easy peeling by lower temperature treatment. PMID- 17112906 TI - Does the presence of unwanted dermal fibroblasts limit the usefulness of autologous epidermal keratinocyte grafts? AB - OBJECTIVES: Fibroblasts sometimes occur after enzymatic isolation of epidermis. They proliferate quickly, overgrowing the culture. A pure epithelial culture is essential for therapy using a keratinocyte graft. The aim of this study was to determine the possibility of fibroblast elimination from culture to prevent fibroblast overgrowth and obtain a pure monolayer of keratinocytes. MATERIAL AND METHODS: We analyzed three epidermal-derived cultures. Cells were cultured in medium contained Dulbecco's Modified Eagle Medium (DMEM) and Ham's F-12 at a 3:1 ratio with 5% autologous serum and additives. The epithelial culture was confirmed using pancytokeratin MMF. If fibroblast like cells were present, they were removed using 0.01% edetate disodium dihydrate (Na2EDDA). This procedure was repeated until we obtained pure primary keratinocyte cultures. RESULTS: Fibroblast detachment was observed after Na2EDDA treatment. The procedure was performed twice and pure primary cultures of keratinocyte were achieved in two cases. These two cultures maintained their epithelial-like morphology and cytokeratin expression. One culture was treated four times with Na2EDDA with no effect; the morphology of the cultures became fibroblast-like with no observed cytokeratin expression. CONCLUSIONS: Unwanted dermal fibroblasts can be separated from primary keratinocyte cultures during the first few days after the isolation. Cocultures of unwanted dermal fibroblasts and epidermal keratinocytes can be reverted to pure keratinocyte monolayers suitable as grafts for transplantation. PMID- 17112907 TI - Allogeneic ovarian orthotopic transplantation in rabbits without a vascular pedicle: morphological, endocrinologic, and natural pregnancy assessment. AB - BACKGROUND: To assess the natural pregnancy of rabbits undergoing bilateral oophorectomy and determine the morphofunctional aspects of orthotopic transplantation of allogeneic intact and sliced ovarian tissue without a vascular pedicle. METHODS: Thirty-two female rabbits had their ovaries removed and orthotopically transplanted between the two breeds of rabbits without vascular anastomoses: group 1 (n = 8), only laparotomy; group 2A (n = 8) intact ovaries transplanted on both sides; group 2B (n = 8), ovaries sliced and orthotopically transplanted; group 2C (n = 8), an intact ovary transplanted on one side and a sliced ovary on the other side. Three months later, the animals were paired with males for copulation. We assessed estradiol, progesterone, follicle-stimulating hormone, and luteinizing hormone levels. A histological study was performed, and the number of pregnancies and litters determined. RESULTS: Pregnancies occurred in seven rabbits of group 1; in 37.5% of rabbits in group 2A; 50% in group 2B; and 62.5% in group 2C. Hormone levels and histology confirmed the vitality of all ovaries. CONCLUSIONS: Intact or sliced orthotopic allogeneic ovarian transplantations without a vascular pedicle are viable in rabbits, preserving hormonal and fertile functions. PMID- 17112908 TI - Refined techniques for intestinal transplantation in rat. AB - Intestinal transplantation (IT), unlike other solid-organ transplantations, such as liver, kidney, and heart, has relatively disappointing results in humans. Significant advances have been made during the past 40 years, but rejection, graft-versus-host disease (GVHD), and infection remain the major obstacles to successful IT. These aspects may be studied using a rat model of IT. Without a microscope and traditional suture for vascular reconstruction, we simplified the procedure using a "three cuffs" technique for orthotopic intestinal transplantation (OIT). Technical modifications of our OIT model that induced good results included (1) adopting a "double cuffs" technique on the graft aorta, (2) using a portal cuff anastomosis to reconstitute the natural and physiologic portal graft drainage with the cuff fixed to the recipient first, and (3) administering a large volume of crystalloid or whole blood to maintain blood pressure and reduce ischemic injury to the graft during operation. In our group, the survival rate of recipients was 87.5% (21 of 24 rats), the average volume of bleeding in the recipient operation was less than 1 mL, and the cold ischemic time, 50 +/- 11 minutes. PMID- 17112909 TI - Prolonged maintenance of neointestine using subcutaneously implanted tubular scaffolds in a rat model. AB - Tissue-engineered small intestine offers a possible alternative to long-term parenteral nutrition or intestinal transplantation in patients with short bowel syndrome. The aim of this study was to investigate the prolonged development of neointestine grown on subcutaneously implanted scaffolds. Tubular polylactide coglycolide (PLGA) scaffolds were implanted into adult Lewis rats. Four weeks after scaffold implantation, a suspension of organoid units was delivered to the lumen of each scaffold. Organoid units were manufactured from small intestine harvested from neonatal Lewis rats by partial digestion using collagenase and dispase. Scaffolds were removed at 4, 8, and 12 weeks after organoid unit implantation, processed to paraffin, and sectioned. Hematoxylin and eosin staining demonstrated well-developed and well-differentiated intestinal mucosa and a vascularised submucosa within the scaffolds at 4, 8, and 12 weeks. Appearances were similar to native small intestine. Immunohistochemistry performed using primary antibody against proliferating cell nuclear antigen, a marker for cellular proliferation, demonstrated positively staining cells within the mucosa and submucosa at all time points. In the mucosal layer these positively staining cells were found primarily in the crypts. These findings show that neointestinal mucosa can be maintained for at least 12 weeks on a subcutaneous PLGA scaffold, and the presence of actively proliferating cells at 12 weeks suggests potential for further development beyond this. PMID- 17112910 TI - Embryonic stem cell derived and adult hematopoietic stem cell transplantation for tolerance induction in a renal allograft recipient:--a case report. AB - We generated an human embryonic stem cell (hESC) line to augment chimerism associated tolerance. A 40-year-old African with chronic glomerulonephritis chronic renal failure with 100% G6PD enzyme deficiency presented for renal transplantation with a 27-year-old, 6/6 HLA-matched sister as a willing donor. METHOD: We generated an hESC line from the donor's oocytes using long ovarian stimulation protocol simultaneously with tolerance induction protocol. A nuclear transfer (NT)-hESC line was derived by transferring a donor cumulus cell into an enucleated oocyte, subjected to electrical fusion, and cultured for 5 days. ESCs hatched from the blastocyst on day 6 were cocultured with her unmodified bone marrow for 2 days and suspended in Ringer's lactate. Five milliliters of suspension were collected for cell counting, viability, pluripotency, flow cytometry, and karyotyping. The remaining suspension was infused into the periphery of the recipient. Transplantation was performed 1 week later following a negative lymphocytotoxicity cross-match test using no immunosuppression. Peripheral blood chimerism (PBC) was studied using fluorescent in situ hybridization technique. Allograft biopsy was performed on day 7. RESULTS: NT hESC CD34+ count was 7.6%, viability 100%, karyotyping normal, pluripotency markers: SSEA-1, SSEA-4, OCT-3/4, TRA-1/60:positive; 12% PBC was noted at 1 week after transplantation. Serum creatinine was 1.2 mg%, graft biopsy was unremarkable, and G6PD enzyme deficiency was corrected to 0% at 100 days posttransplant. Liver function tests and hematology profile were unremarkable for graft-versus-host disease. CONCLUSION: This is the first report of tolerance induction using NT-hESC-induced hematopoietic chimerism with synergistic use of adult bone marrow. It was safe and effective. PMID- 17112911 TI - Complications of laparoscopic gastric banding in renal transplant recipients: a case study. AB - As bariatric surgery becomes more popular, the number of renal transplant recipients who undergo weight loss surgery will continue to grow. This population presents unique challenges because of increased infection risks, tendency to posttransplant weight gain, and inferior tissue-healing properties. We present two cases of renal transplant recipients who experienced the complications of band erosion and band migration after laparoscopic gastric banding, and we discuss the special considerations that apply to this patient population. PMID- 17112912 TI - Severe hyperkalemic type 4 renal tubular acidosis after kidney transplantation: a case report. AB - BACKGROUND: Hyperkalemia after transplantation is a common event, occurring in up to 70% of patients. It is usually asymptomatic but sometimes manifests as muscle weakness or cardiac arrhythmias. METHODS: Case report. RESULTS: At 102 days after a second cadaveric kidney transplantation, a 15-year-old boy, was admitted to the emergency room with severe muscle weakness. His examinations showed a serum potassium of 9.8 mEq/L; blood pH 7.1; serum bicarbonate 7.6 mmol/L; and creatinine 2.5 mg/dL. He was initially treated with sodium bicarbonate, calcium gluconate, and furosemide. Subsequent investigation showed hyperchloremic metabolic acidosis, urinary pH <5.5, positive urinary anion gap, reduced transtubular potassium gradient (TTKG, 1.5) and low levels of aldosterone (0.7 ng/mL), suggesting the presence of type 4 renal tubular acidosis (RTA). Other causes of hyperkalemia were excluded in the present case. Serum levels of potassium returned to normal when fludrocortisone was added to the bicarbonate supplementation. This case of severe hyperkalemic secondary to type 4 RTA after kidney transplantation only responded to the combination of alkali and mineralocorticoid therapies. PMID- 17112913 TI - Acute cytomegalovirus infection complicated by venous thrombosis in a renal transplant recipient. AB - We report a case of simultaneous acute cytomegalovirus infection and venous thrombosis in a renal transplant recipient. On posttransplant month 3, the patient started complaining of left leg pain and swelling. Tibiopopliteal and femoral deep venous thrombosis were confirmed by Doppler ultrasonography. A serological test for CMV ELISA was strongly positive for IgM antibodies. Acute CMV infection was diagnosed by serum quantitative DNA polymerase chain reaction. Genetic predisposing risk factors for thrombosis (eg, protein C and S deficiency, factor V Leiden and prothrombin G20210A mutations, and antithrombin III deficiency) were not present. Results of tests for anticardiolipin antibodies, lupus anticoagulant, and antinuclear antibodies were also negative. No other clinical or biologic risk factors for thrombosis were detected in the patient. The patient responded well to intravenous gancyclovir and low-molecular weight heparin therapy. He was discharged in good condition. Our observation suggests that acute CMV infection may be the cause of a thrombotic event in renal transplant recipients. PMID- 17112914 TI - Treatment of lamivudine-resistant hepatitis B infection in post-renal transplant patients with adefovir dipivoxil: preliminary results. AB - We report the treatment outcome in six post-renal transplant patients with lamivudine-resistant hepatitis B virus (LR-HBV) infection using adefovir dipivoxil and followed for 6 to 18 months. Posttransplant immunosuppressive therapy was not altered. Adefovir dipivoxil effectively suppressed hepatitis B virus DNA and improved alanine transferase, although DNA suppression seemed dependent on continued therapy. Nephrotoxicity led to withdrawal of the drug in three patients. This may limit therapeutic usefulness in a significant proportion of post-renal transplant patients with LR-HBV infection. PMID- 17112915 TI - A case of isolated Nocardia asteroides brain abscess in a kidney transplant recipient. AB - Because of the immunosuppressive drugs used after organ transplantation, there is an increased rate of certain infections and malignancies. Nocardia brain abscess is a rare condition, seen most commonly among immunocompromised patients. It may be confused with intracranial tumors and requires long-term combined antibiotic therapy after drainage. CASE REPORT: A patient who underwent renal transplantation because of end-stage renal disease of unknown origin was shown to have a nocardial brain abscess while she was taking immunosuppressive drugs. The patient was given combined antibiotics and the abscess drained surgically. After 2 months, antibiotic therapy was continued with one drug. Neither a complication nor clinical or radiological sequelae occurred in this patient. CONCLUSIONS: When central nervous system findings are observed in renal transplant recipients, nocardial brain abscess must be considered in the differential diagnosis. Recommended treatment duration is 6 to 12 months with frequent imaging. PMID- 17112916 TI - Lamivudine reverses severe acute hepatitis B and pancytopenia after renal transplantation: a case report. AB - Pancytopenia is rare after acute hepatitis B infection. The use of lamivudine in the treatment of acute hepatitis B-associated pancytopenia in renal transplant recipients has not been documented. Herein we reported a 21-year-old woman who was infected with acute hepatitis B 6 months after renal transplantation, a condition complicated by pancytopenia. Lamivudine reversed the acute hepatitis in 1 month and the pancytopenia after 3 months, without a change in renal function. Lamivudine was maintained for 2 years without a hepatitis flare-up after 4 years. PMID- 17112917 TI - Intraarterial thrombolytic treatment for hepatic artery thrombosis immediately after living donor liver transplantation. AB - Hepatic artery thrombosis (HAT) following living donor liver transplantation (LDLT) remains one of the major causes of graft failure and mortality in liver transplant recipients. This complication requires early diagnosis and revascularization to avoid graft loss. We have reported herein two cases of successful urokinase intraarterial thrombolytic treatment for HAT in the immediate postoperative period after LDLT. Significant elevation of liver transaminases was noted 6 and 4 hours after LDLT and HAT confirmed by three dimensional computed tomogram and angiogram. Both patients were treated successfully with intraarterial thrombolysis using an urokinase infusion (a total dose of 200,000 to 250,000 IU over 20 to 25 minutes) immediately after HAT was confirmed. One patient underwent laparotomy and bleeder ligation owing to hepatic arterial anastomotic site bleeding after thrombolysis. These two patients remain in good condition without any ischemic graft sequelae at 7 and 8 months follow up. In conclusion, intraarterial thrombolysis using an urokinase infusion could be considered as one of the treatment modalities of acute HAT following LDLT even in the immediate postoperative period. PMID- 17112918 TI - Malaria in a liver transplant recipient: a case report. AB - Malaria is an exotic complication in liver transplants patients. It can be acquired either by transfusion of blood products or through the transplanted organ. Infections caused by Plasmodium spp are unusual in liver transplants; to date, only four cases have been reported in the literature. Herein we have presented a case of Plasmodium vivax in a liver transplant patient. This diagnosis must be excluded in febrile transplant patients in endemic areas, especially during the first 2 months. An epidemiological history relevant for malaria both in the donor and in the recipient must be routinely included with screening tests. PMID- 17112919 TI - Favorable resolution of hepatic infarctions in transplanted liver after portal vein thrombosis treated by surgical thrombectomy: a case report. AB - Portal vein thrombosis (PVT) after orthotopic liver transplantation (OLT) is a life-threatening complication associated with a high rate of graft loss and patient death, with reported incidence of 1% to 2% in adults. We report a case of an early PVT after OLT complicated by hepatic infarctions in the liver graft. After surgical thrombectomy and restoration of the portal inflow, hepatic infarctions resolved spontaneously within 6 months, which was confirmed by computed tomography. PMID- 17112920 TI - Transient ischemic attack after rizatriptan administration in a liver transplant recipient: a case report. AB - We report the case of a male liver transplant recipient who developed de novo migraine while on tacrolimus therapy. Considering the inadequate control of pain using nonsteroidal antiinflammatory drugs, rizatriptan benzoate (10 mg orally) was administered (double administration). After both administrations a clinically transient ischemic attack (TIA) occurred. Rizatriptan was discontinued, the patient recovered without sequelae from both episodes of TIA. Remission of migraine occurred after discontinuation of tacrolimus and substitution with cyclosporine. We suggest that the association of rizatriptan and tacrolimus could potentially lead to an excessive risk of cerebral vasospasm and should be used with caution. A change in immunosuppressive therapy (from tacrolimus to cyclosporine or sirolimus) may improve migraine and should be the first choice. Further prospective comparative randomized trials are needed to establish the best therapeutic option in this particular subset of patients. PMID- 17112921 TI - Very late recurrence of hepatocellular carcinoma after liver transplantation: case report and literature review. AB - Hepatocellular carcinoma (HCC) recurs in 10% to 60% of the patients after liver transplantation (OLT) and is associated with increased mortality. The average time to recurrence ranges from 1 to 2 years following OLT, and the median survival from the time of diagnosis is about 1 year. We report a case of a 69 year-old man who underwent OLT for hepatitis C virus-related cirrhosis with HCC, and was diagnosed with recurrent HCC 6.5 years after OLT. Biopsies from the initial and recurrent tumors showed a well-differentiated HCC with foci of clear cell pattern. The patient was still alive and asymptomatic 32 months after the diagnosis despite extensive tumor burden. He expired 9 years, 9 months after OLT and 3 years, 2 months after the detection of recurrence. In conclusion, HCC may recur more than 6 years after OLT and may exhibit an indolent course. This case illustrates the highly variable rate of tumor growth and progression post-OLT. The impact of this information on the need for long-term surveillance for recurrent HCC post-OLT remains to be determined. PMID- 17112922 TI - Acute appendicitis in a kidney-pancreas transplant recipient: report of a case. AB - The prompt diagnosis and management of acute surgical conditions in immunocompromised solid organ transplant recipients are of critical importance. These conditions may or may not be related or to the transplanted allograft(s). This is a case report of a 41-year-old woman who received a simultaneous pancreas kidney transplant. Nine years after the transplant, she developed acute appendicitis with a periappendiceal abscess and a fecalith, and she was treated with percutaneous drainage of the abscess and eventual semielective appendectomy. This is the first known report of acute appendicitis in a pancreas allograft recipient in the English literature. PMID- 17112923 TI - Diabetic muscle infarction after kidney and pancreas transplantation: case report and literature review. AB - Diabetic muscle infarction (DMI) is a rare, long-term complication of poorly controlled diabetes (typically of type I). DMI was first described in 1965 and more than 100 cases have been reported thereafter in the English literature. Usually, there is a coexistence with concomitant nephropathy, neuropathy, and retinopathy. The etiology remains uncertain, but appears to be attributable to diabetic microangiopathy and hypercoagulability and is believed that hypoxia reperfusion injury is involved. DMI presents with sudden onset of pain associated with a tender mass in the thigh in most instances. The diagnosis is based on magnetic resonance imaging, which is not specific but highly indicative. Treatment is conservative with relapses occurring in 50% of the patients, but not necessarily in the same muscle group. We describe a case of DMI that occurred 4 months after simultaneous kidney and pancreas transplantation in one patient with type I diabetes mellitus and end-stage renal disease. PMID- 17112924 TI - Recurrent lymphangioleiomyomatosis after living-donor lobar lung transplantation. AB - Living-donor lobar lung transplantation (LDLLT) has been applied to patients with various end-stage lung diseases. The recurrence of pulmonary lymphangioleiomyomatosis (LAM) after lung transplantation has been rarely reported. Herein, we report a case of recurrent pulmonary LAM after LDLLT. A 24 year-old woman presented with pneumothorax and infiltrates in the left lung 1 year after bilateral LDLLT for LAM. These symptoms and radiologic findings occurred repeatedly and then improved quickly. Thereafter, computed tomography of the chest revealed a tiny emphysematous change of the subpleural region in the left lung, which was exacerbated gradually and was finally diagnosed as LAM recurrence by transbronchial lung biopsy. In previous reports of LAM recurrence, the diagnosis was made at the time of autopsy. This is also the first reported case diagnosed early, that is, when the patient was alive and her allograft had not deteriorated badly. PMID- 17112926 TI - The risk of missed diagnosis of acute myocardial infarction associated with emergency department volume. AB - STUDY OBJECTIVE: Missed diagnosis of acute myocardial infarction is associated with adverse clinical outcomes and more dollars recovered in malpractice suits than any other condition. The rate of missed diagnosis varies between emergency departments (EDs); we hypothesized that it is associated with the volume of acute myocardial infarction patients treated in an ED and that the association can be explained by other hospital characteristics. METHODS: We linked the records of all acute myocardial infarction patients admitted to an Ontario hospital in 2002 to 2003 to their ED visit records in the 7 days preceding admission. Acute myocardial infarctions were defined as missed if the diagnosis on the previous visit matched a list of cardiac symptoms and illnesses. We assessed whether annual volume of admitted acute myocardial infarction patients treated in the ED (grouped as 0 to 49; 50 to 99; 100 to 199; 200 to 299; and > or = 300) was associated with missed acute myocardial infarction, adjusting for age, sex, teaching hospital status, and acute myocardial infarction severity. In a secondary analysis, we used data from a survey of Ontario EDs to assess whether hospital characteristics (triage practices, use of diagnostic tests, and consultant availability) explained the volume association. RESULTS: Of 19,663 acute myocardial infarction patients, mean age (68.3 years), sex (63% men), and predicted 1-year mortality (mean 0.21; SD 0.18) were similar across volume groups. The rate of missed acute myocardial infarction was 2.1% (95% confidence interval [CI] 1.9% to 2.3%) and varied from 0% to 29% across EDs. Compared with very high-volume EDs, the adjusted odds ratio of missed acute myocardial infarction was 2.0 in very low- (95% CI 1.5 to 2.7) and 1.6 in low- (95% CI 1.1 to 2.3) volume EDs. Consultant availability partially explained the volume effect. CONCLUSION: Lower-volume EDs have up to 2-fold higher odds of missed acute myocardial infarctions compared with highest-volume ones after controlling for patient factors. Many current technologies designed to increase diagnostic sensitivity are feasible only in higher-volume centers. Efforts to reduce overall rates of missed acute myocardial infarctions should instead focus on simpler solutions appropriate for lower-volume EDs, such as telemedicine to improve access to consultant expertise. PMID- 17112927 TI - Images in emergency medicine. Extensive groin abscess and myositis after intravenous cubital buprenorphine injection. PMID- 17112928 TI - Volume and outcome: the more patients the better? PMID- 17112930 TI - Discordant cardiac biomarkers: frequency and outcomes in emergency department patients with chest pain. AB - STUDY OBJECTIVE: We evaluate associations between pairs of discordant cardiac biomarkers (positive MB band of creatine kinase [CKMB] with negative creatine kinase, positive CKMB with negative cardiac troponin, and positive troponin with negative CKMB) and the presence of acute coronary syndromes in emergency department (ED) chest pain patients. METHODS: This was a secondary analysis of a prospective registry. Data were obtained from the multicenter Internet Tracking Registry of Acute Coronary Syndromes, which included 17,713 ED visits for possible acute coronary syndrome between June 1999 and August 2001. First visits and first ED cardiac biomarker results from the 9 sites, 8 in the United States and 1 in Singapore, were included. Subjects were excluded for incomplete information or an initial ECG consistent with ST-segment elevation myocardial infarction. Acute coronary syndrome was defined by diagnosis-related group code indicating myocardial infarction, positive invasive or noninvasive diagnostic testing, revascularization, or death during hospitalization or within 30 days. RESULTS: Of 8,769 eligible patients, 1,614 (18.4%) had acute coronary syndrome. The CKMB and cardiac troponin results were discordant in 7% of patients (CKMB+/cardiac troponin-, 4.9%, CKMB-/cardiac troponin+ 2.1%), whereas increased CKMB with normal creatine kinase levels occurred in 239 (3.1%) patients. The unadjusted odds ratios with 95% confidence intervals for acute coronary syndrome in patients with and without discordant markers were: CKMB+/CK- 5.7 (4.4-7.4), CKMB+/CK+ 4.4 (3.6-5.2), CKMB-/cTn+ 4.8 (3.4-6.8), CKMB+/cTn- 2.2 (1.7-2.8), CKMB+/cTn+ 26.6 (18.0-39.3). For the group with cardiac troponin, the reference category was negative troponin and negative CKMB; for the group with creatine kinase, the reference category was negative CKMB but either a positive or negative creatine kinase. CONCLUSION: Among the spectrum of ED patients with chest pain, an increased CKMB level with a normal creatine kinase level identifies patients at increased risk for acute coronary syndrome. Similarly, an increased troponin level regardless of CKMB level and an increased CKMB level regardless of troponin level identify patients at higher risk for acute coronary syndrome than those with uniformly normal cardiac biomarker levels. Our data suggest that discordant cardiac biomarkers may identify patients at increased risk for acute coronary syndrome. PMID- 17112931 TI - Diagnosis of lower limb deep venous thrombosis in emergency department patients: performance of Hamilton and modified Wells scores. AB - STUDY OBJECTIVE: We validate and compare the Hamilton score for assessment of lower limb deep venous thrombosis with the modified Wells score. METHODS: Consecutive patients presenting to the emergency department of a tertiary center for suspected lower limb deep venous thrombosis were prospectively recruited. Hamilton score and modified Wells score calculations, D-dimer, and complete (calf veins included), single lower limb ultrasonographic examination were performed for all patients. All patients with a negative ultrasonographic examination result for deep venous thrombosis were followed up for 3 months. RESULTS: The study population consisted of 116 men and 193 women, with an average age of 55.6 years (SD 20.1). A total of 67 (21.7%) patients were diagnosed with deep venous thrombosis. Forty (59.7%) of these patients had isolated calf deep venous thrombosis, and the other 27 (40.3%) patients had proximal deep venous thrombosis. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (LR+), and negative likelihood ratio (LR-) for an unlikely Hamilton score (score < or = 2) and D dimer were 99% (95% confidence interval [CI] 92% to 99.96%), 42 % (95% CI 36% to 49%), 32% (95% CI 26% to 39%), 99% (95% CI 95% to 99.98%), 1.7 (95% CI 1.52% to 1.9%), and 0.04 (95% CI 0.01 to 0.25), respectively. The sensitivity, specificity, PPV, NPV, LR+ and LR- for an unlikely modified Wells score (score < or = 1) and D-dimer were 99% (95% CI 92% to 99.96%), 33 % (95% CI 27% to 39%), 29% (95% CI 23% to 35%), 99% (95% CI 93% to 99.97%), 1.47 (95% CI 1.34 to 1.62), and 0.05 (95% CI 0.01 to 0.32), respectively. CONCLUSION: An unlikely probability of Hamilton score and a negative D-dimer may effectively exclude a lower limb deep venous thrombosis. Hamilton and modified Wells scores have similar performance characteristics. PMID- 17112932 TI - Patients who leave without being seen: their characteristics and history of emergency department use. AB - STUDY OBJECTIVE: We identify patient characteristics associated with uncompleted visits to the emergency department (ED). METHODS: We used registration and billing data to conduct a pair-matched case-control study. ED patients who left without being seen (cases) between July 1 and December 31, 2004, were matched to patients who stayed and were treated (N=1,476 pairs) according to registration date and time (+/-2 hours) and triage level (controls). The association between sociodemographic characteristics, previous ED utilization, and proximity to the ED and the risk of an uncompleted visit was assessed by the odds ratio (OR) using conditional logistic regression. RESULTS: During the 6-month study period, the overall left-without-being-seen rate was 6.4%. Seventeen percent of cases compared with 5% of controls had at least 1 previous uncompleted visit during the previous year. After adjusting for all patient characteristics, younger age, being uninsured (adjusted OR=1.73; 95% confidence interval [CI] 1.35 to 2.21) or covered by Medicaid (adjusted OR=1.67; 95% CI 1.27 to 2.20), and a previous uncompleted visit (adjusted OR=3.60; 95% CI 2.67 to 4.85) were significantly associated with the risk of an uncompleted visit. CONCLUSION: Previous ED utilization is predictive of future ED utilization. EDs should make every effort to keep their left-without-being-seen rates to a minimum because patients who are the least likely to receive care elsewhere (ie, those uninsured or covered by Medicaid) are more likely to leave without being seen. PMID- 17112933 TI - Ambulance diversion and lost hospital revenues. AB - STUDY OBJECTIVE: We estimate ambulance revenues lost from each hour spent on ambulance diversion at an urban teaching hospital's emergency department (ED) and examine the financial impact of increased ICU capacity, which reduced diversion hours by 63%. METHODS: This was a secondary analysis of administrative data to determine the time and date of ambulance arrivals, as well as the insurance status and revenues from each ED patient arriving by ambulance between January 1, 2002, and December 31, 2003. The primary outcome measure was hourly revenues (ie, payments to the hospital) for ambulance patients. RESULTS: Ten thousand three hundred one adult, non-trauma-system ED patients arrived by ambulance in 2002 and 2003, with average hospital revenues of 4,492 dollars. Each hour spent on diversion was associated with 1,086 dollars (95% confidence interval 611 dollars to 1,461 dollars) in forgone hospital revenues from ambulance patients. In August 2002, the study hospital increased its staffed ICU beds from 47 to 67, and diversion decreased from an average of 307 to 114 hours per month. In association with the reduction in diversion, the hospital received more patients by ambulance, which translated into approximately 175,000 dollars in additional monthly revenues from ambulance patients. However, these gains were relatively small in relation to total ambulance revenues and to their large monthly variance. CONCLUSION: Ambulance patients generated substantial revenues for hospital services. Decreasing diversion time led to improved revenues. The potential for increased revenues may provide some incentive for hospitals to take greater efforts to reduce ambulance diversion. PMID- 17112934 TI - Ambulance diversion: economic and policy considerations. PMID- 17112935 TI - Clinical decisionmaking: opening the black box of cognitive reasoning. PMID- 17112936 TI - Critical thinking and decisionmaking: avoiding the perils of thin-slicing. PMID- 17112937 TI - Commentary: Morphing social norms through media messaging. PMID- 17112938 TI - The content of medical journal Instructions for authors. AB - STUDY OBJECTIVE: We describe the general and statistical content of the Instructions for Authors of major medical journals. METHODS: This article reports on 2 observational studies. In study 1, we investigated the online versions of Instructions for Authors of 166 journals from 33 specialties for the presence of content about 15 methodologic and statistical topics. In study 2, we categorized the general content of the online versions of the Instructions for Authors of 35 medical journals. Two abstractors independently assigned the content into 18 categories and counted the total number of words devoted to each category. Interrater reliability of the classification was assessed. RESULTS: Less than half of the 166 Instructions for Authors in study 1 provided any guidance on statistical methods, and the majority failed to cite accepted publication standards such as the International Committee of Medical Journal Editors Uniform Guidelines or CONSORT. Only 13% of journals commented on the content and style of data tables and figures. The 35 Instructions for Authors in study 2 varied greatly in length (mean 3,308; median 2,283; range 885 to 18,927) and, with few exceptions, focused on formatting issues. Forty-three percent of Instructions offered no advice on scientific content, and only 5 journals devoted more than 10% of their words to scientific content. CONCLUSION: There is great heterogeneity among medical journal Instructions for Authors. Instructions provide little guidance about methodologic and statistical issues, and the advice provided is often contradictory among journals. PMID- 17112939 TI - Unintended lessons from the veterinarian. PMID- 17112940 TI - Images in emergency medicine. Isolated proximal tibiofibular dislocation. PMID- 17112941 TI - Thirty-day versus 7-day outcomes in the San Francisco Syncope Rule. PMID- 17112943 TI - ATLS versus ETC: time for a decision? PMID- 17112945 TI - Symposium on the definition and management of anaphylaxis. PMID- 17112946 TI - Medical decisionmaking and the San Francisco Syncope Rule. PMID- 17112949 TI - 40-MHz annular array imaging of mouse embryos. AB - Ultrasound biomicroscopy (UBM) has emerged as an important in vivo imaging approach for analyzing normal and genetically engineered mouse embryos. Current UBM systems use fixed-focus transducers, which are limited in depth-of-focus. Depending on the gestational age of the embryo, regions-of-interest in the image can extend well beyond the depth-of-focus for a fixed-focus transducer. This shortcoming makes it particularly problematic to analyze 3-D data sets and to generate accurate volumetric renderings of the mouse embryonic anatomy. To address this problem, we have developed a five-element, 40-MHz annular array transducer and a computer-controlled system to acquire and reconstruct fixed- and array-focused images of mouse embryos. Both qualitative and quantitative comparisons showed significant improvement with array-focusing, including an increase of 3 to 9 dB in signal-to-noise ratio and an increase of at least 2.5 mm in depth-of-focus. Volumetric-rendered images of brain ventricles demonstrated the clear superiority of array-focusing for 3-D analysis of mouse embryonic anatomy. PMID- 17112950 TI - Examination of cancer in mouse models using high-frequency quantitative ultrasound. AB - Two mouse models of mammary cancer (a carcinoma and sarcoma) were examined using quantitative ultrasound (QUS). Scatterer property estimates, i.e., the average scatterer diameter (ASD) and average acoustic concentration (AAC), were estimated from regions-of-interest (ROIs) inside the tumors. Initially, the spherical Gaussian model was used over an analysis bandwidth of 10 to 25 MHz to obtain ASD and AAC estimates. ASD estimates were 31.7 +/- 9.36 microm and 31.0 +/- 7.20 microm for the carcinomas and sarcomas, respectively. AAC estimates were 6.77 +/- 8.75 dB[mm(-3)] and 9.87 +/- 9.24 dB[mm(-3)], respectively. The initial ASD and AAC estimates did not yield statistically significant differences between the two kinds of tumors (p = 0.83, 0.86 for the ASD and AAC estimates, respectively). However, optical photomicrographs revealed distinct morphologic differences between the tumors. F-tests on the average power spectra from the tumors revealed statistically significant differences between the spectra over the range of 16 to 25 MHz. ASD and AAC estimates using the spherical Gaussian model were then obtained over the new analysis bandwidth of 16 to 25 MHz. The new ASD estimates were 42.1 +/- 4.01 microm and 32.1 +/- 3.81 microm for the carcinomas and sarcomas, respectively. The new AAC estimates were 16.4 +/- 17.1 dB[mm(-3)] and 36.4 +/- 11.9 dB[mm(-3)], respectively. Statistically significant differences were observed for both the ASD and AAC estimates when using the new analysis bandwidth. Structural differences between the tumors were revealed by both QUS and optical photomicrographs. PMID- 17112951 TI - Harmonic intravascular ultrasound imaging with a dual-frequency catheter. AB - Recent studies have shown the feasibility of tissue and contrast harmonic imaging with a prototype nonlinear intravascular ultrasound (IVUS) system using a conventional single-element rotating IVUS catheter. In this study, a dual frequency transducer element was mounted in an IVUS catheter and its second harmonic imaging performance was investigated and compared with that of a conventional IVUS catheter. Hydrophone measurements showed a transmit efficiency improvement of >6 dB for the dual-frequency catheter at 20 MHz. In vitro phantom experiments showed a signal-to noise ratio improvement of >5 dB in second harmonic mode at 40 MHz (H40) with the dual-frequency catheter, when using equal transmit voltage for both catheters. Finally, in vivo experiments were conducted and showed image improvement in H40 acquisitions with respect to the conventional IVUS catheter. PMID- 17112952 TI - Measurement of wave velocity in arterial walls with ultrasound transducers. AB - Arterial wall stiffness can be associated with various diseases. The stiffness of an artery can be measured with the pulse wave velocity (PWV) using the "foot-to foot" method. However, the foot of the pressure pulse is not very clear, due to reflected waves. The blood pressure pulse generated by the heart is a low frequency wave and its time resolution is low. PWV is an average indicator of artery stiffness between the two measuring positions; therefore, it cannot easily identify local stiffness. In this paper, a sinusoidally modulated force with a high frequency is generated noninvasively on the arterial wall by the radiation force of ultrasound (US). The resulting vibration in the artery is measured with an US Doppler transceiver. The wave velocity in the artery is measured from a wave image obtained by scanning the force transducer and fixing the sensor transducer. Because of the high imposed force frequency, the temporal resolution of this method is much higher than the conventional pressure PWV method. Local wave velocity more than a few millimeters can be measured, which is not possible with the PWV method. PMID- 17112953 TI - Noninvasive simultaneous assessment of wall shear rate and wall distension in carotid arteries. AB - A novel technique has been developed for the noninvasive real-time simultaneous assessment of both blood velocity profile and wall displacements in human arteries. The novel technique is based on the use of two ultrasound beams, one set at optimal angle for wall motion measurements and the other for blood velocity profile measurements. The technique was implemented on a linear array probe divided into two subapertures. A modified commercial ultrasound machine and a custom PC board based on a high-speed digital signal processor was used to process the quadrature demodulated echo signals and display results in realtime. Flow phantom experiments demonstrated the validity of the technique, providing wall shear rate (WSR) estimates within 10% of the theoretical values. The system was also tested in the common carotid arteries of 16 healthy volunteers (age 30 to 53 y). Results of simultaneous diameter distension and WSR measurements were in agreement with published data. PMID- 17112954 TI - On the statistics of ultrasonic spectral parameters. AB - Several factors affect the accuracy and precision of ultrasonic spectrum analysis, which is used for characterization of normal and diseased tissue in a variety of organs. For example, averaging procedures and the sequence of operations affect the accuracy and precision of spectrum analysis. Averaging procedures and logarithmic conversion (i.e., conversion to dB) introduce a constant bias that affects spectral amplitudes and the values of intercept and midband fit; the bias depends on the sequencing of the log conversion and averaging as well as the number of independent spectra or spectral parameters that are averaged. We derive expressions that permit correction of such biases. Furthermore, we show that standard deviations for slope and midband-fit estimation can be minimized by averaging spectra before dB conversion and before computing spectral parameters by linear regression. Experimental results using phantoms agree remarkably with theoretical predictions for the data window functions studied in this article, Hamming and rectangular. PMID- 17112955 TI - Technique for rapid in vitro single-cell elastography. AB - Statistically meaningful assays require scanning a large number of cells in a short time. Here we present a technique for rapid in vitro single-cell elastography. The technique is based on atomic force acoustic microscopy but (1) requires only a few minutes of scan time, (2) can be used on live cells briefly removed from most of the nutrient fluid, (3) does negligible harm or damage to the cell, (4) provides semiquantitative information on the distribution of modulus across the cell and (5) yields data with 1- to 10-nm resolution. We describe the new technique in detail, apply it to baby hamster kidney cells, verify the results and calibrate the images with atomic force microscope force distance measurements. The technique enables rapid assessment of physical/biochemical signals on the cell modulus and contributes to current understanding of cell mechanics. PMID- 17112956 TI - ARFI imaging for noninvasive material characterization of atherosclerosis. AB - Cardiovascular disease (CVD) is the leading cause of death in the United States, with 70% of CVD mortalities the result of sequelae of atherosclerosis. An urgent need for enhanced delineation of vulnerable plaques has catalyzed the development of novel atherosclerosis imaging strategies that use X-ray computed tomography, magnetic resonance and ultrasound modalities. As suggested by the pathophysiology of plaque development and progression to vulnerability, insight to the focal material, i.e., mechanical, properties of arterial walls and plaques may enhance atherosclerosis characterization. We present acoustic radiation force impulse (ARFI) ultrasound in application to mechanically characterizing a raised focal atherosclerotic plaque in an iliac artery extracted from a relevant pig model. ARFI results are correlated to matched immunohistochemistry, indicating elastin and collagen composition. In regions of degraded elastin, slower recovery rates from peak ARFI-induced displacements were observed. In regions of collagen deposition, lower ARFI-induced displacements were achieved. This work demonstrates ARFI for characterizing the material nature of an atherosclerotic plaque. PMID- 17112957 TI - Model-based imaging. AB - The concept of model-based imaging provides a possibility of integration of both structural and functional information obtained by imaging and nonimaging sources of diagnostically relevant information. Merging information of different origin and nature in a simulated computer patient-specific three-dimensional (3-D) model is important for multimodal imaging approaches to medical diagnostics. The work of F. L. Lizzi on life-like imaging done nearly 20 years ago was the first successful attempt of patient-specific 3-D computer modeling using conventional ultrasonography data. He demonstrated the applications of such 3-D models, which incorporated acoustic, optical and thermal properties of imaged tissue, in physiologic studies, in planning and monitoring ultrasonic hyperthermia and ablation. However, numerous obstacles hinder the wide use of the model-based imaging concept. Using the example of model-based imaging of prostate, the advantages and limitation on the applicability of the concept are discussed. Attempts of implementing the model-based imaging concept in the mechanical imaging technology--imaging with the use of measurements of stress pattern on the surface of tissue--are described. It is shown that composing a patient-specific 3 D model requires well-defined and carefully validated algorithms for translating features of the object assessed by an imaging modality into the quantitative anatomic and histopathologic parameters. PMID- 17112958 TI - Biological and physical mechanisms of HIFU-induced hyperecho in ultrasound images. AB - Guidance and monitoring of high intensity focused ultrasound (HIFU) therapy, using ultrasound imaging, has primarily utilized formation of a hyperechoic region at the HIFU focus. We investigated biologic and physical mechanisms of a hyperecho, as well as safety of this phenomenon, using thermal, acoustic and light microscopy observations. Single, short-duration HIFU pulses (30-60 ms) were able to produce a hyperechoic region at the HIFU focus, 2 cm deep in a rabbit thigh muscle. When hyperechoic regions appeared, inertial cavitation was detected in vivo using a custom-made passive cavitation detection system. Light micrographs showed a large number of cavities (approximately 100/mm3), 1-10 microm in diameter, in a cytoplasm of cells located at the HIFU focus. Blood congestion was observed around a focal region, indicating an injury of microvasculature. Cellular necrosis was observed at 2 d after the treatment, while healing, scar tissue formation and regeneration were observed at 7 d and 14 d. The results indicate that a possibility of adverse tissue effects has to be taken into consideration when the hyperecho formation, induced by very-short HIFU pulses, is used for pretreatment targeting. PMID- 17112959 TI - Use of a bovine eye lens for observation of HIFU-induced lesions in real-time. AB - Study of coagulative lesion formation by high intensity focused ultrasound (HIFU) in tissue usually requires performing a sequence of experiments under different exposure conditions followed by tissue sectioning. This paper, inspired by the pioneering work of Frederic L. Lizzi, reports on the use of the bovine eye lens as a laboratory model to observe visually the development of HIFU-induced lesions. The first part of this work describes the measurement of the lens shape, density, sound speed and attenuation. The measured values were within the range of previously published values. In the second part, HIFU-induced lesion development was observed in real-time and compared with good agreement with theoretical simulation. Theoretical modeling included acoustic propagation, absorptive heating and thermal dose, as well as the experimentally measured lens characteristics. Thus, the transparent eye lens can be used as a laboratory phantom to facilitate the understanding of HIFU treatment in other tissues. PMID- 17112960 TI - Improved visualization of high-intensity focused ultrasound lesions. AB - Spectral parameter imaging in both the fundamental and harmonic of backscattered radio-frequency (RF) data were used for immediate visualization of high-intensity focused ultrasound (HIFU) lesion sites. A focused 5-MHz HIFU transducer with a coaxial 9-MHz focused single-element diagnostic transducer was used to create and scan lesions in chicken breast and freshly excised rabbit liver. B-mode images derived from the backscattered RF signal envelope were compared with midband fit (MBF) spectral parameter images in the fundamental (9-MHz) and harmonic (18-MHz) bands of the diagnostic probe. Images of HIFU-induced lesions derived from the MBF to the calibrated spectrum showed improved contrast (approximately 3 dB) of tumor margins versus surround compared with images produced from the conventional signal envelope. MBF parameter images produced from the harmonic band showed higher contrast in attenuated structures (core, shadow) compared with either the conventional envelope (3.3 dB core; 11.6 dB shadow) or MBF images of the fundamental band (4.4 dB core; 7.4 dB shadow). The gradient between the lesion and surround was 3.4 dB/mm, 6.9 dB/mm and 17.2 dB/mm for B-mode, MBF-fundamental mode and MBF-harmonic mode, respectively. Images of threshold and "popcorn" lesions produced in freshly excised rabbit liver were most easily visualized and boundaries best-defined using MBF-harmonic mode. PMID- 17112961 TI - The use of a segmented transducer for rib sparing in HIFU treatments. AB - The use of focused ultrasound as a minimally invasive treatment for tumours is rapidly expanding. Target organs include the liver and kidneys. Both single element and phased array transducers may be used in the clinic. The presence of the rib cage presents a problem in high intensity focused ultrasound (HIFU) treatment planning, due to its high attenuation of the HIFU beam resulting in a loss of power at the focus as well as an increase in the risk of damage at the rib and to overlying tissues, including the skin. In this paper, a linearly segmented transducer, in which all active elements are driven in phase, has been investigated. The aim of the study was to investigate how a beam with a clinically useful profile could be achieved by removing the contribution of edge segments from one side of the transducer to the field. We have considered the case in which the HIFU beam approaches the rib cage during a treatment and investigated configurations of the transducer for which up to three segments on the edge are switched off. This problem has been studied initially using a linear acoustic field program to model the segmented transducer's acoustic beam profile. Experimental measurements of the transducer's acoustic field were performed using an automated beam plotting system. Temperature measurements were made on a rib surface for two transducer configurations using a fine wire thermocouple. A thermochromic liquid crystal material was used to assess qualitatively the heating pattern generated by the ultrasound beam. We show the rib sparing potential of the segmented transducer during HIFU treatment by demonstrating a reduction in the prefocal width of the ultrasound beam when edge segments are switched off. This has been predicted with the acoustic field model and demonstrated experimentally by acoustic field measurements and observations of the heating pattern generated by the ultrasound beam. A significant decrease in the temperature rise on a rib was observed in the case for which three edge segments were switched off compared with when all segments were active. We conclude that a segmented transducer extends the potential for treating liver tumours. In the case where the tumour lies behind, but close to the edge of, the ribs, energy loss at the focus and excessive heating in the rib and overlying tissue can be avoided by switching off edge segments. PMID- 17112962 TI - Lesions of ultrasound-induced lung hemorrhage are not consistent with thermal injury. AB - Thermal injury, a potential mechanism of ultrasound-induced lung hemorrhage, was studied by comparing lesions induced by an infrared laser (a tissue-heating source) with those induced by pulsed ultrasound. A 600-mW continuous-wave CO2 laser (wavelength approximately 10.6 microm) was focused (680-microm beamwidth) on the surface of the lungs of rats for a duration between 10 to 40 s; ultrasound beamwidths were between 310 and 930 microm. After exposure, lungs were examined grossly and then processed for microscopic evaluation. Grossly, lesions induced by laser were somewhat similar to those induced by ultrasound; however, microscopically, they were dissimilar. Grossly, lesions were oval, red to dark red and extended into subjacent tissue to form a cone. The surface was elevated, but the center of the laser-induced lesions was often depressed. Microscopically, the laser-induced injury consisted of coagulation of tissue, cells and fluids, whereas injury induced by ultrasound consisted solely of alveolar hemorrhage. These results suggest that ultrasound-induced lung injury is most likely not caused by a thermal mechanism. PMID- 17112963 TI - Therapeutic effects of paclitaxel-containing ultrasound contrast agents. AB - Drug delivery vehicles that combine ultrasonic and molecular targeting are shown to locally concentrate a drug in a region-of-interest. The drug delivery vehicles, referred to as acoustically active lipospheres (AALs), are microbubbles surrounded by a shell of oil and lipid. In a region limited to the focal area of ultrasound application, circulating AALs are deflected by radiation force to a vessel wall and can subsequently be fragmented. Ligands targeting the alphavbeta3 integrin are conjugated to the AAL shell and increase in vitro binding by 26.5 fold over nontargeted agents. Toxicity assays demonstrate that paclitaxel containing AALs exert a greater antiproliferative effect after insonation than free paclitaxel at an equivalent concentration. Lastly, ultrasound and molecular targeting are combined to deliver a model drug to the endothelium and interstitium of chorioallantoic membrane vasculature in vivo. PMID- 17112964 TI - Vascular lesions and s-thrombomodulin concentrations from auricular arteries of rabbits infused with microbubble contrast agent and exposed to pulsed ultrasound. AB - Arterial injury resulting from the interaction of contrast agent (CA) with ultrasound (US) was studied in rabbit auricular arteries and assessed by histopathologic evaluation and s-thrombomodulin concentrations. Three sites on each artery were exposed (2.8 MHz, 5-min exposure duration, 10-Hz pulse repetition frequency, 1.4-mus pulse duration) using one of three in situ peak rarefactional pressures (0.85, 3.9 or 9.5 MPa). Saline, saline/CA, and saline/US infusion groups (n = 28) did not have histopathologic damage. The saline/CA/US infusion group (n = 10) at exposure conditions below the FDA mechanical index limit of 1.9 did not have histopathologic damage, whereas the saline/CA/US infusion group (n = 9) at exposure conditions above the FDA limit did have damage (5 of 9 arteries). Lesions were characteristic of acute coagulative necrosis. Mean s-thrombomodulin concentrations, a marker for endothelial cell injury, were highest in rabbits exposed to US at 0.85 and 3.9 MPa, suggesting that vascular injury may be physiological and not accompanied by irreversible cellular injury. PMID- 17112965 TI - Threshold estimation of ultrasound-induced lung hemorrhage in adult rabbits and comparison of thresholds in mice, rats, rabbits and pigs. AB - The objective of this study was to assess the threshold and superthreshold behavior of ultrasound (US)-induced lung hemorrhage in adult rabbits to gain greater understanding about species dependency. A total of 99 76 +/- 7.6-d-old 2.4 +/- 0.14-kg New Zealand White rabbits were used. Exposure conditions were 5.6 MHz, 10-s exposure duration, 1-kHz PRF and 1.1-micros pulse duration. The in situ (at the pleural surface) peak rarefactional pressure, p(r(in situ)), ranged between 1.5 and 8.4 MPa, with nine acoustic US exposure groups plus a sham exposure group. Rabbits were assigned randomly to the 10 groups, each with 10 rabbits, except for one group that had nine rabbits. Rabbits were exposed bilaterally with the order of exposure (left then right lung, or right then left lung) and acoustic pressure both randomized. Individuals involved in animal handling, exposure and lesion scoring were blinded to the exposure condition. Probit regression analysis was used to examine the dependence of the lesion occurrence on in situ peak rarefactional pressure and order of exposure (first vs. second). Likewise, lesion depth and lesion root surface area were analyzed using Gaussian tobit regression analysis. Neither probability of a lesion nor lesion size measurements was found to be statistically dependent on the order of exposure after the effect of p(r(in situ)) was considered. Also, a significant correlation was not detected between the two exposed lung sides on the same rabbit in either lesion occurrence or size measures. The p(r(in situ)) threshold estimates (in MPa) were similar to each other across occurrence (3.54 +/- 0.78), depth (3.36 +/- 0.73) and surface area (3.43 +/- 0.77) of lesions. Using the same experimental techniques and statistical approach, great consistency of thresholds was demonstrated across three species (mouse, rat and rabbit). Further, there were no differences in the biologic mechanism of injury induced by US and US induced lesions were similar in morphology in all species and age groups studied. The extent of US-induced lung damage and the ability of the lung to heal led to the conclusion that, although US can produce lung damage at clinical levels, the degree of damage does not appear to be a significant medical problem. PMID- 17112966 TI - Application of TILLING in plant improvement. AB - TILLING (Targeting induced local lesions in genomes) is a general reverse-genetic strategy that is used to locate an allelic series of induced point mutations in genes of interest. High-throughput TILLING allows the rapid and cost-effective detection of induced point mutations in populations of chemically mutagenized individuals. The technique can be applied not only to model organisms but also to economically important organisms in plants. Owing to its full of advantages such as simple procedure, high sensitivity, and high efficiency, TILLING provides a powerful approach for gene discovery, DNA polymorphism assessment, and plant improvement. Coupled with other genomic resources, TILLING and EcoTILLING can be used immediately as a haplotyping tool in plant breeding for identifying allelic variation in genes exhibiting expression correlating with phenotypes and establishing an allelic series at genetic loci for the traits of interest in germplasm or induced mutants. PMID- 17112967 TI - Comparative analysis of complete mitochondrial DNA control region of four species of Strigiformes. AB - The sequence of the whole mitochondrial (mt) DNA control region (CR) of four species of Strigiformes was obtained. Length of the CR was 3,290 bp, 2,848 bp, 2,444 bp, and 1,771 bp for Asio flammeus, Asio otus, Athene noctua, and Strix aluco, respectively. Interestingly, the length of the control region was maximum in Asio flammeus among all the avian mtDNA control regions sequenced thus far. In addition, the base composition and organization of mtDNA CR of Asio flammeus were identical to those reported for other birds. On the basis of the differential frequencies of base substitutions, the CR may be divided two variable domains, I and III, and a central conserved domain, II. The 3' end of the CR contained many tandem repeats of varying lengths and repeat numbers. In Asio flammeus, the repeated sequences consisted of a 126 bp sequence that was repeated seven times and a 78 bp sequence that was repeated 14 times. In Asio otus, there were also two repeated sequences, namely a 127 bp sequence that was repeated eight times and a 78 bp sequence that was repeated six times. The control region of Athene noctua contained three sets of repeats: a 89 bp sequence that was repeated three times, a 77 bp sequence that was repeated four times, and a 71 bp sequence that was repeated six times. Strix aluco, however, had only one repeated sequence, a 78 bp sequence that was repeated five times. The results of this study seem to indicate that these tandem repeats may have resulted from slipped-strand mispairing during mtDNA replication. Moreover, there are many conserved motifs within the repeated units. These sequences could form stable stem-loop secondary structures, which suggests that these repeated sequences play an important role in regulating transcription and replication of the mitochondrial genome. PMID- 17112968 TI - Cloning and characterization of Bombyx mori PP-BP a gene induced by viral infection. AB - The ENF peptide family, so termed after the consensus sequence in their amino termini (Glu-Asn-Phe-), is assumed to play multiple important roles in defense reactions, growth regulation, and homeostasis of Lepidopteran insects. The paralytic peptide of Bombyx mori (BmPP) is one such peptide that is involved in the paralytic and plasmatocyte-spreading activities in the hemocyte immune reaction. The growth-blocking peptide of Pseudaletia separata (PsGBP), which is also a member of the ENF peptide family, has similar functions that can reportedly be attenuated by the growth-blocking peptide-binding protein (GBP-BP). Using the fluorescent differential display (FDD) technique, the differential expression pattern of genes in highly susceptible silkworm strain 306 were analyzed, following infection with B. mori nuclear polyhedrosis virus (BmNPV), and a differential band (G12(782)) was obtained from the hemolymph RNA pools. Using 5'-RACE with a specially designed primer based on the FDD study, a 1,401 bp cDNA clone was obtained containing a 1,311 bp open reading frame (ORF, GenBank accession number DQ306881). The deduced protein was highly homologous in primary structure to GBP-BP and was termed B. mori paralytic peptide-binding protein (PP BP). The B. mori PP-BP gene is organized into two exons and only one intron, using bioinformatics searches.Using RT-PCR analysis, it was found that the B. mori PP-BP gene was expressed almost exclusively in the hemolymph. Real-time quantitative PCR analysis indicated that the B. mori PP-BP mRNA level in B. mori strain 306 exposed to BmNPV was much higher than that in B. mori strain without the virus infection. This result implies that the B. mori PP-BP is related to the cellular immune response after BmNPV invades the hemolymph. PMID- 17112969 TI - Analysis of expressed sequence tags from skeletal muscle-specific cDNA library of Chinese native Xiang pig. AB - A Longissimus Dorsi muscle cDNA library of Xiang Pig was constructed, and 131 randomly isolated clones were sequenced in this study. The results of bioinformatics analysis showed that 131 ESTs represented 109 unique clones sequences, of which 99 showed homology to previously identified genes in humans or other mammals, 3 matched other uncharacterized expressed sequence tags (ESTs), and 7 showed no significant matches to sequences already present in DNA databases. No protein matches were found for 10 ESTs. Functional analysis of the ESTs showed that a considerable proportion of them encoded proteins involved in gene/protein expression (45.46%). Other classes included genes involved in metabolism (10.10%), cell structure/motility (10.10%), cell/organism defense (5.05%), cell signaling/communication (2.02%), and cell division (0.0%). Unclassified genes constituted the remaining 27.27%. This study reported the results of the first gene expression profile analysis of Chinese native Xiang Pig skeletal muscle cells, thereby greatly facilitating the functional study of candidate genes involved in muscle growth as well as in the improvement of meat quality in domestic pigs. PMID- 17112970 TI - Effects of the MyoG gene on the partial growth traits in pigs. AB - Single-nucleotide polymorphisms of the MyoG gene were tested using PCR-SSCP in different pig breeds including Landrace, Large White, Duroc, Shanxi Black, and Mashen pigs, and the effects of the MyoG gene on the birth weight, the weaning weight, the 6-month body weight, and the backfat thickness were also analyzed. On the basis of the published sequence of the porcine MyoG gene, ten pairs of primers were designed, and one polymorphism was found in the PCR product amplified with In2-3 primers. The results showed that: (1) the Landrace, the Large White, and the Duroc breeds differ significantly (P < 0.05) in genotype distribution from the Shanxi Black and the Mashen breeds; (2) On the basis of the fixed effect model, significant differences were found in the birth weight and the backfat thickness among the different MyoG genotypes, whereas no significant differences existed in the weaning weight and the 6-month body weight; (3) Using least square analysis, it was seen that individuals of the BB genotype had significantly less (P < 0.01) birth weight than those of the AA and AB genotypes, with the order being AA>AB>BB; the pigs of the AA genotype had significantly lower (P < 0.01) backfat thickness than those of the AB and BB genotypes, with the order being AA50% lumen narrowing) was compared with quantitative coronary angiography. All segments were analyzed regardless of image quality from coronary calcification or motion artifacts. Results were analyzed by patient and by coronary segment (990) using the American Heart Association 15-segment model. RESULTS: Lower heart rates were associated with improved image quality. Computed tomography correctly identified 35 of 37 (95%) patients without significant stenosis and 28 of 29 (97%) patients with significant stenosis on CCA. Computed tomography correctly assessed 68 of 94 (72%) significant stenosis. Overall, accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of 64-slice CT for identifying CAD by patient was 95%, 97%, 95%, 93%, and 97%, respectively, and by segment was 97%, 72%, 99%, 91%, and 97%, respectively. CONCLUSIONS: In a routine clinical practice, 64-slice CT detects with excellent accuracy a significant CAD in patients with complete LBBB. A normal CT in this clinical setting is a robust tool to act as a filter and avoid invasive diagnostic procedures. PMID- 17112980 TI - Detecting coronary artery disease in left bundle branch block. PMID- 17112981 TI - In vivo analysis of the anatomical relationship of coronary sinus to mitral annulus and left circumflex coronary artery using cardiac multidetector computed tomography: implications for percutaneous coronary sinus mitral annuloplasty. AB - OBJECTIVES: We sought to determine the in vivo anatomical relationships between mitral annulus (MA) and coronary sinus (CS) as well as CS and left circumflex coronary artery using cardiac computed tomography. BACKGROUND: Percutaneous treatment of mitral regurgitation (MR) by annuloplasty via CS is under development. Success of such treatment depends on the close anatomical proximity of the MA to the CS. The in vivo data regarding this anatomical relationship in humans are scant. We investigated this relationship using contrast multidetector computed tomography. METHODS: We studied 25 normal individuals and 11 patients with severe MR (3 to 4+) due to mitral valve prolapse. Separation between MA and CS was measured in standard planes, in 4-chamber (4C), 2-chamber (2C), and 3 chamber views. Distance from ostium of CS to the intersection with left circumflex (LCX), and anatomical relation of LCX and CS were determined using 3 dimensional mapping (Philips Brilliance, Philips Medical Systems, Amsterdam, the Netherlands). RESULTS: There was significant variance of CS to MA separation at all planes. Separation of CS and MA was increased in lateral location (4C) and decreased in posterior location (2C) in the MR group with increase in MA size. Left circumflex artery crossed between CS and MA in 80% of patients. The LCX crossed CS at a variable distance from the ostium of CS (86.5 +/- 21 mm, range 37 to 123 mm) CONCLUSIONS: There is significant variability in the relation of CS to MA in humans. Coronary sinus to MA distance increases in patients with severe MR and annular dilation, mainly in the posterolateral location. The left circumflex crosses under the CS the majority of times, but with a significant variability in the location where it crosses the CS. These anatomical features should be taken into consideration while selecting percutaneous treatment strategies for mitral valve repair. PMID- 17112982 TI - Detection of coronary artery stenosis with whole-heart coronary magnetic resonance angiography. AB - OBJECTIVES: We sought to determine the diagnostic performance of whole-heart coronary magnetic resonance (MR) angiography for detecting significant coronary artery disease. BACKGROUND: The accuracy of whole-heart coronary MR angiography has not been determined in a large number of patients. METHODS: Three-dimensional coronary MR angiograms covering the entire heart were obtained during free breathing in 131 patients. Images were acquired during a patient-specific time window in the cardiac cycle with minimal motion of the coronary artery. Significant coronary artery disease was defined on X-ray coronary angiography as a diameter reduction of > or =50% in coronary arteries with a reference diameter of > or =2 mm. RESULTS: The acquisition of MR angiography was completed in 113 (86%) of 131 patients, with an imaging time averaged at 12.9 +/- 4.3 min. On a patient-based analysis, the sensitivity, specificity, positive and negative predictive value, and accuracy of MR angiography were 82% (95% confidence interval [CI] 69% to 91%), 90% (95% CI 79% to 96%), 88% (95% CI 74% to 95%), 86% (95% CI 75% to 93%), and 87% (95% CI 79% to 92%), respectively. These values in the individual segments were 78% (95% CI 68% to 85%), 96% (95% CI 95% to 97%), 69% (95% CI 60% to 77%), 98% (95% CI 96% to 98%), and 94% (95% CI 96% to 96%). CONCLUSIONS: Whole-heart coronary MR angiography allows for noninvasive detection of significant narrowing in coronary arterial segments with a diameter of > or =2 mm with moderate sensitivity and high specificity. PMID- 17112983 TI - Coronary MRI: more pretty pictures or present-day value? PMID- 17112984 TI - Delayed enhancement magnetic resonance imaging predicts response to cardiac resynchronization therapy in patients with intraventricular dyssynchrony. AB - OBJECTIVES: We evaluated the ability of delayed enhancement magnetic resonance imaging (DE-MRI) to predict clinical response to cardiac resynchronization therapy (CRT). BACKGROUND: Cardiac resynchronization therapy reduces morbidity and mortality in selected heart failure patients. However, up to 30% of patients do not have a response. We hypothesized that scar burden on DE-MRI predicts response to CRT. METHODS: The DE-MRI was performed on 28 heart failure patients undergoing CRT. Patients with QRS > or =120 ms, left ventricular ejection fraction < or =35%, New York Heart Association functional class II to IV, and dyssynchrony > or =60 ms were studied. Baseline and 3-month clinical follow-up, wall motion, 6-min walk, and quality of life assessment were performed. The DE MRI was performed 10 min after 0.20 mmol/kg intravenous gadolinium. Scar measured by planimetry was correlated with response criteria. RESULTS: Twenty-three patients completed the protocol (mean age 64.9 +/- 11.7 years), with 12 (52%) having a history of myocardial infarction. Thirteen (57%) patients met response criteria. Percent total scar was significantly higher in the nonresponse versus response group (median and interquartile range of 24.7% [18.1 to 48.7] vs. 1.0% [0.0 to 8.7], p = 0.0022) and predicted nonresponse by receiver-operating characteristic analysis (area = 0.94). At a cutoff value of 15%, percent total scar provided a sensitivity and specificity of 85% and 90%, respectively, for clinical response to CRT. Similarly, septal scar < or =40% provided a 100% sensitivity and specificity for response. Regression analysis showed linear correlations between percent total scar and change in each of the individual response criteria. CONCLUSIONS: The DE-MRI accurately predicted clinical response to CRT. This technique offers unique information in the assessment of patients referred for CRT. PMID- 17112985 TI - Discrimination of myocardial acute and chronic (scar) infarctions on delayed contrast enhanced magnetic resonance imaging with intravascular magnetic resonance contrast media. AB - OBJECTIVES: The purpose of this study was to examine the potential of intravascular gadolinium (Gd)-chelates in discriminating acute from chronic myocardial infarctions (MIs). BACKGROUND: A potential limitation of delayed contrast enhanced magnetic resonance imaging with standard extracellular Gd chelates is its inability to distinguish acute from chronic MIs. METHODS: Eight pigs with MIs were studied at 3 days and 8 weeks. Inversion recovery gradient echo (IR-GRE), T(1)-turbo spin echo (TSE), and T(2)-TSE images were acquired before and after administration of intravascular and extracellular Gd-chelates. Triphenyltetrazolium chloride (TTC) was used to delineate infarctions at postmortem. Masson's trichrome and Biotinylated Bandeiria simplicifolia Isolectin B4 stains were used to characterize scarred myocardium. Analysis of variance was used to compare signal intensity (SI) ratios and determine differences in infarct extent. RESULTS: The intravascular agent produced differential enhancement of acute infarctions at 3 days (SI ratio 5.8 +/- 1.3) but not at 8 weeks (1.6 +/- 0.4, p < 0.01). The extracellular agent provided differential enhancement of both acute (SI ratio 7.7 +/- 1.4) and chronic (7.5 +/- 0.9) infarctions. The extents of enhanced regions in acute infarctions were not different after intravascular (16.0 +/- 1.3%) or extracellular (17.1 +/- 1.7%) agents; at 8 weeks the extent of extracellular enhanced and TTC regions were smaller (13.2 +/- 1.4% and 12.0 +/- 1.5%, respectively). Masson's trichrome stain demonstrated dense scar tissue, signaling the complete healing of infarction. The vascular stain showed that scar tissue contained fewer microvessels oriented in a haphazard array. CONCLUSIONS: The combination of intravascular and extracellular Gd-chelates discriminates acute from chronic infarctions on delayed images. This double contrast agent approach can be used to determine the age and extent of infarctions. PMID- 17112986 TI - Contrast-enhanced cardiovascular magnetic resonance imaging of right ventricular infarction. AB - OBJECTIVES: We assessed the role of late enhancement cardiovascular magnetic resonance imaging (LE-CMR) for the diagnosis of right ventricular infarction (RVI). BACKGROUND: Right ventricular infarction occurs in about one-half of patients with inferior myocardial infarction (MI). It is associated with an unfavorable prognosis, but established methods often lack the diagnostic accuracy to detect it. Late enhancement cardiovascular magnetic resonance imaging accurately detects left ventricular MI. METHODS: Thirty-seven patients with acute inferior MI were included. To test for RVI, they prospectively underwent a physical examination, an electrocardiogram (ECG) for ST-segment elevation in the V4r right precordial lead, and an echocardiogram. After coronary reperfusion, LE CMR was performed for assessing presence and extent of late enhancement in the right ventricular (RV) wall. The LE-CMR data were compared with the other results; interobserver variability was assessed. The LE-CMR was repeated after 13 months. RESULTS: Late enhancement cardiovascular magnetic resonance imaging detected RVI in 21 of 37 (57%) patients with acute inferior MI. Interobserver variability was very good (kappa 0.83); physical exam was positive for RVI in 7 of 37 (19%) patients, V4r ECG in 13 of 37 (35%) patients, and echocardiogram in 6 of 37 (16%) patients. The LE-CMR findings for RVI showed only mild agreement with findings for RVI on physical exam (kappa 0.30), V(4)r ECG (kappa 0.38), and echocardiography (kappa 0.32). Irreversible injury of the RV persisted at 13 months (kappa 0.85). CONCLUSIONS: In patients with acute inferior MI, RVI is more frequently detected by LE-CMR than by current standard diagnostic techniques. Further CMR studies might allow for analyzing its clinical and prognostic relevance. PMID- 17112987 TI - Cardiovascular magnetic resonance, fibrosis, and prognosis in dilated cardiomyopathy. AB - OBJECTIVES: We studied the prognostic implications of midwall fibrosis in dilated cardiomyopathy (DCM) in a prospective longitudinal study. BACKGROUND: Risk stratification of patients with nonischemic DCM in the era of device implantation is problematic. Approximately 30% of patients with DCM have midwall fibrosis as detected by late gadolinium-enhancement (LGE) cardiovascular magnetic resonance (CMR), which may increase susceptibility to arrhythmia and progression of heart failure. METHODS: Consecutive DCM patients (n = 101) with the presence or absence of midwall fibrosis were followed up prospectively for 658 +/- 355 days for events. RESULTS: Midwall fibrosis was present in 35% of patients and was associated with a higher rate of the predefined primary combined end point of all cause death and hospitalization for a cardiovascular event (hazard ratio 3.4, p = 0.01). Multivariate analysis showed midwall fibrosis as the sole significant predictor of death or hospitalization. However, there was no significant difference in all-cause mortality between the 2 groups. Midwall fibrosis also predicted secondary outcome measures of sudden cardiac death (SCD) or ventricular tachycardia (VT) (hazard ratio 5.2, p = 0.03). Midwall fibrosis remained predictive of SCD/VT after correction for baseline differences in left ventricular ejection fraction between the 2 groups. CONCLUSIONS: In DCM, midwall fibrosis determined by CMR is a predictor of the combined end point of all-cause mortality and cardiovascular hospitalization, which is independent of ventricular remodeling. In addition, midwall fibrosis by CMR predicts SCD/VT. This suggests a potential role for CMR in the risk stratification of patients with DCM, which may have value in determining the need for device therapy. PMID- 17112988 TI - The expanding prognostic role of late gadolinium enhanced cardiac magnetic resonance. PMID- 17112989 TI - Left ventricular structure and function: basic science for cardiac imaging. AB - The myofiber geometry of the left ventricle (LV) changes gradually from a right handed helix in the subendocardium to a left-handed helix in the subepicardium. In this review, we associate the LV myofiber architecture with emerging concepts of the electromechanical sequence in a beating heart. We discuss: 1) the morphogenesis and anatomical arrangement of muscle fibers in the adult LV; 2) the sequence of depolarization and repolarization; 3) the physiological inhomogeneity of transmural myocardial mechanics and the apex-to-base sequence of longitudinal and circumferential deformation; 4) the sequence of LV rotation; and 5) the link between LV deformation and the intracavitary flow direction observed during each phase of the cardiac cycle. Integrating the LV structure with electrical activation and motion sequences observed in vivo provides an understanding about the spatiotemporal sequence of regional myocardial performance that is essential for noninvasive cardiac imaging. PMID- 17112990 TI - Myocardial strain and torsion quantified by cardiovascular magnetic resonance tissue tagging: studies in normal and impaired left ventricular function. AB - Accurate quantification and timing of regional myocardial function allows early identification of dysfunction, and therefore becomes increasingly important for clinical risk assessment, patient management, and evaluation of therapeutic efficacy. For this purpose, the application of tissue Doppler echocardiography has rapidly increased. However, echocardiography has some major inherent limitations. Cardiovascular magnetic resonance imaging with tissue tagging provides highly reproducible data on myocardial function, not only in longitudinal and radial directions, but also in the circumferential direction. Because of the development of faster imaging protocols, improved temporal resolution, less time-consuming postprocessing procedures, and the potential of quantifying myocardial deformation in 3 dimensions at any point in the heart, this technique may serve as an alternative for tissue Doppler echocardiography and is now ready for more widespread clinical use. This review discusses the clinical use of cardiovascular magnetic resonance tissue tagging for quantitative assessment of regional myocardial function, thereby underlining the specific features and emerging role of this technique. PMID- 17112991 TI - Assessment of left ventricular function by cardiac ultrasound. AB - Our understanding of the physical underpinnings of the assessment of cardiac function is becoming increasingly sophisticated. Recent developments in cardiac ultrasound permit exploitation of many of these newer physical concepts with current echocardiographic machines. This review will first focus on the current approach to the assessment of cardiovascular hemodynamics by cardiac ultrasound. The next focus will be the assessment of global cardiac mechanics in systole and diastole. Finally, relationships between the cardiac structure and regional myocardial function, and the way regional function can be quantified by ultrasound, will be presented. This review also discusses the clinical impact of echocardiography and its future directions and developments. PMID- 17112992 TI - Differentiation of subendocardial and transmural infarction using two-dimensional strain rate imaging to assess short-axis and long-axis myocardial function. AB - OBJECTIVES: This study sought to differentiate the transmural extent of infarction (TME) by assessment of the short-axis and long-axis function of the left ventricle (LV) using 2-dimensional (2D) strain. BACKGROUND: The differentiation of subendocardial infarction from transmural infarction has significant prognostic and clinical implications. METHODS: Contrast-enhanced magnetic resonance imaging (CE-MRI) and dobutamine stress echocardiography (DBE) were performed in 80 patients (age 63 +/- 10 years) with chronic ischemic LV dysfunction. Myocardial function was assessed in the short axis at the midventricular level using peak strain rate (SR) and strain (S) in circumferential and radial dimensions, and was assessed in the long axis using longitudinal SR and S. Wall motion analysis was performed during DBE to assess for contractile reserve. RESULTS: Transmural infarct segments had lower circumferential S (-10.7 +/- 6.3) and SR (-1.0 +/- 0.4) than subendocardial infarcts (S: -15.4 +/- 7.0, p < 0.0001; SR: -1.4 +/- 0.8, p = 0.02) and normal myocardium (S: p < 0.0001; SR: p < 0.0001). Transmural and subendocardial infarct segments had similar radial S and SR. Subendocardial infarct segments showed significant reduction of longitudinal S (-13.2 +/- 5.6) and SR (-0.91 +/- 0.45) compared with normal myocardium (S: -17.8 +/- 5.4, p < 0.0001; SR: -1.1 +/- 0.41, p < 0.0001), but there were no significant differences between subendocardial and transmural infarct segments (p = 0.09). Wall motion analysis by DBE could not identify subendocardial infarction on CE-MRI (TME 1% to 50%: DBE scar 38%, DBE viable 38%, DBE ischemic 24%, p = NS). CONCLUSIONS: The combined assessment of long-axis and short-axis function using 2D strain may be used to identify TME. PMID- 17112993 TI - Evaluation of global and regional left ventricular function with 16-slice computed tomography, biplane cineventriculography, and two-dimensional transthoracic echocardiography: comparison with magnetic resonance imaging. AB - OBJECTIVES: We sought to compare left ventricular (LV) function assessed with multislice computed tomography (MSCT), biplane cineventriculography (CVG), and transthoracic echocardiography (Echo), with magnetic resonance imaging (MRI) as the reference standard. BACKGROUND: With the same data as acquired for noninvasive coronary angiography, MSCT enables registration of myocardial function. METHODS: A total of 88 patients (64 men and 24 women) underwent MSCT with 16 x 0.5 mm detector collimation, CVG, and MRI, whereas Echo was retrospectively analyzed in a subset of 30 patients. RESULTS: Regarding the ejection fraction, the agreement was significantly superior for MSCT than for CVG (+/- 10.2% vs. +/- 16.8%; p < 0.001) and Echo (+/- 11.0% vs. +/- 21.2%; p < 0.001). For the end-diastolic and end-systolic volumes, the limits of agreement with CVG (p < 0.001) and Echo (p < 0.001 and p < 0.02, respectively) were also significantly larger than with MSCT. In comparison with MSCT, CVG significantly overestimated the end-diastolic and end-systolic volumes (p < 0.001). Intraobserver analysis of MSCT yielded limits of agreement for ejection fraction (+/- 4.8%), end-diastolic volume (+/- 15.6 ml) and end-systolic volume (+/- 8.0 ml), and myocardial mass (+/- 18.2 g). The accuracy in identifying patients and myocardial segments with abnormal regional function was significantly higher with MSCT (84% and 95%) than with CVG (63% and 90%; p < 0.002 and p < 0.001), whereas MSCT and Echo were not significantly different in identifying patients with abnormal regional function. CONCLUSIONS: Our results indicate that the assessment of global and regional LV function with MSCT is more accurate than with CVG, whereas MSCT is superior to Echo for global function. This suggests that MSCT allows reliable evaluation of global and regional LV function. PMID- 17112994 TI - Single-beat noninvasive imaging of cardiac electrophysiology of ventricular pre excitation. AB - OBJECTIVES: The aim of this study was to determine whether noninvasive imaging of cardiac electrophysiology (NICE) is feasible in patients with Wolff-Parkinson White (WPW) syndrome in the clinical setting of a catheter laboratory and to test the accuracy of the noninvasively obtained ventricular activation sequences as compared with that of standard invasive electroanatomic mapping. BACKGROUND: NICE of ventricular activation could serve as a useful tool in the treatment of cardiac arrhythmias and might help improve our understanding of arrhythmia mechanisms. METHODS: NICE works by fusing the data from high-resolution electrocardiographic mapping and a model of the patient's cardiac anatomy obtained by magnetic resonance imaging. The ventricular activation sequence was computed with a bidomain theory-based heart model to solve this inverse problem. Noninvasive imaging of cardiac electrophysiology was performed in 7 patients with WPW syndrome undergoing catheter ablation of the accessory pathway. The position error of NICE was defined as the distance between the site of earliest activation computed by NICE and the successful ablation site identified by electroanatomic mapping (CARTO; Biosense Webster, Diamond Bar, California) for normal atrioventricular (AV) conduction as well as for adenosine-induced AV block. RESULTS: The error introduced by geometric coupling of the CARTO data and the NICE model was 5 +/- 3 mm (model discretization 10 mm). All ventricular accessory pathway insertion sites were identified with an accuracy of 18.7 +/- 5.8 mm (baseline) and 18.7 +/- 6.4 mm (adenosine). CONCLUSIONS: The individual cardiac anatomy model obtained for each patient enables accurate noninvasive electrocardiographic imaging of ventricular pre-excitation in patients with WPW syndrome. Noninvasive imaging of cardiac electrophysiology might be used as a complementary noninvasive approach to localize the origin and help identify and understand the underlying mechanisms of cardiac arrhythmias. PMID- 17112995 TI - Three-dimensional echocardiography: the benefits of the additional dimension. AB - Over the past 3 decades, echocardiography has become a major diagnostic tool in the arsenal of clinical cardiology for real-time imaging of cardiac dynamics. More and more, cardiologists' decisions are based on images created from ultrasound wave reflections. From the time ultrasound imaging technology provided the first insight into the human heart, our diagnostic capabilities have increased exponentially as a result of our growing knowledge and developing technology. One of the most significant developments of the last decades was the introduction of 3-dimensional (3D) imaging and its evolution from slow and labor intense off-line reconstruction to real-time volumetric imaging. While continuing its meteoric rise instigated by constant technological refinements and continuing increase in computing power, this tool is guaranteed to be integrated in routine clinical practice. The major proven advantage of this technique is the improvement in the accuracy of the echocardiographic evaluation of cardiac chamber volumes, which is achieved by eliminating the need for geometric modeling and the errors caused by foreshortened views. Another benefit of 3D imaging is the realistic and unique comprehensive views of cardiac valves and congenital abnormalities. In addition, 3D imaging is extremely useful in the intraoperative and postoperative settings because it allows immediate feedback on the effectiveness of surgical interventions. In this article, we review the published reports that have provided the scientific basis for the clinical use of 3D ultrasound imaging of the heart and discuss its potential future applications. PMID- 17112996 TI - Transesophageal real-time three-dimensional echocardiography methods and initial in vitro and human in vivo studies. AB - OBJECTIVES: The purpose of this study was to develop a transesophageal probe that: 1) enables on-line representation of the spatial structures of the heart, and 2) enables navigation of medical instruments. BACKGROUND: Whereas transthoracic real-time 3-dimensional (3D) echocardiography could recently be implemented, there is still no corresponding transesophageal system. Transesophageal real-time 3D echocardiography would have great potential for numerous clinical applications, such as navigation of catheters. METHODS: The newly developed real-time 3D system is based on a transesophageal probe in which multiple transducers are arranged in an interlaced pattern on a rotating cylinder. This enables continuous recording of a large echo volume of 70 mm in length and a sector angle of 120 degrees . The presentation of the volume reconstructed data is made with a time lag of <100 ms. The frame rate is up to 20 Hz. In addition to conventional imaging, the observer can obtain a stereoscopic image of the structures examined with red/blue goggles. RESULTS: It was shown in vitro on ventricle- and aorta-form agar models and in vivo that the system enables excellent visualization of the 3D structures. Shape, spatial orientation, and the navigation of various catheters (e.g., EPS-catheter, Swan-Ganz-catheter), stents, or atrial septal defect occluders could be recorded on-line and stereoscopically depicted. The size of the echo sector enables a wide field of view without changing the position of the probe. CONCLUSIONS: Transesophageal real-time 3D echocardiography can be technically realized with the system presented here. The in vitro and in vivo studies show particularly the potential for navigation in the heart and large vessels on the basis of stereoscopic images. PMID- 17112997 TI - Cardiovascular imaging in the management of atrial fibrillation. AB - Atrial fibrillation (AF) is he most commonly encountered arrhythmia in clinical practice, with an overall prevalence of 0.4% in the general population. Recent advances in technology and in the understanding of the pathophysiology of AF have led to more definitive and potentially curative therapeutic approaches. Echocardiography has a well-established role in the assessment of cardiac structure and function and risk stratification, and has become an essential part of the guidelines for management of AF. The development of intracardiac echocardiography has led to real-time guidance of percutaneous interventions, including radiofrequency ablation and left atrial appendage closure procedures for patients with AF. Other imaging modalities, including computed tomography and magnetic resonance angiography, have allowed for more accurate measurement and better understanding of the cardiac anatomy. We review the impact of various imaging modalities in the evaluation and management of AF. PMID- 17112998 TI - Noninvasive imaging in myocarditis. AB - Increased recognition of the role of inflammation in acute and chronic dilated cardiomyopathy has revived an interest in noninvasive imaging for detection of myocarditis. Diagnostic strategies that are based on molecular imaging promise to further advance our understanding and improve diagnostic precision. This article reviews the strengths and limitations of common clinical tests used for the diagnosis of myocarditis, with a focus on the emerging role of cardiovascular magnetic resonance imaging. Novel imaging modalities that are currently in preclinical development are discussed with recommendations for future clinical research. PMID- 17113000 TI - Noninvasive characterization of myocardial molecular interventions by integrated positron emission tomography and computed tomography. AB - OBJECTIVES: We sought to investigate the usefulness of integrated positron emission tomography (PET) and computed tomography (CT) for in vivo characterization of an angiogenesis-directed molecular intervention. BACKGROUND: Controversies about the effectiveness of molecular therapies for cardiovascular disease have prompted the need for more powerful noninvasive imaging techniques. METHODS: In a model of regional adenoviral transfer of the VEGF(121) gene to myocardium of healthy pigs, PET-CT using multiple molecular-directed radiotracers was employed. RESULTS: Two days after gene transfer, successful transgene expression was noninvasively confirmed by a reporter probe targeting co-expressed HSV1-sr39tk reporter gene. The CT-derived ventricular function and morphology remained unaltered (left ventricular ejection fraction 57 +/- 5% in adenovirus injected animals vs. 53 +/- 5% in controls; p = 0.36). Increased regional perfusion was identified in areas overexpressing VEGF (myocardial blood flow during adenosine-induced vasodilation 1.47 +/- 0.49 vs. 1.14 +/- 0.27 ml/g/min in remote areas; p = 0.01), corroborating in vivo effects on microvascular tone and permeability. Finally, regional angiogenesis-associated alpha(v)beta3 integrin expression was not enhanced, suggesting little contribution to the perfusion increase. Fusion of CT morphology and tracer-derived molecular signals allowed for accurate regional localization of biologic signals. Findings were validated by control vectors, sham-operated animals, and ex vivo tissue analysis. CONCLUSIONS: Integrated PET-CT has the potential to dissect cardiovascular biologic mechanisms from gene expression to physiologic function and morphology. The VEGF overexpression in healthy myocardium increases myocardial perfusion without significant up-regulation of alpha(v)beta3 integrin adhesion molecules early after the intervention. PMID- 17113001 TI - Multimodality noninvasive imaging demonstrates in vivo cardiac regeneration after mesenchymal stem cell therapy. AB - OBJECTIVES: The purpose of this study was to test the hypothesis, with noninvasive multimodality imaging, that allogeneic mesenchymal stem cells (MSCs) produce and/or stimulate active cardiac regeneration in vivo after myocardial infarction (MI). BACKGROUND: Although intramyocardial injection of allogeneic MSCs improves global cardiac function after MI, the mechanism(s) underlying this phenomenon are incompletely understood. METHODS: We employed magnetic resonance imaging (MRI) and multi-detector computed tomography (MDCT) imaging in MSC treated pigs (n = 10) and control subjects (n = 12) serially for a 2-month period after anterior MI. A sub-endocardial rim of tissue, demonstrated with MDCT, was assessed for regional contraction with MRI tagging. Rim thickness was also measured on gross pathological specimens, to confirm the findings of the MDCT imaging, and the size of cardiomyocytes was measured in the sub-endocardial rim and the non-infarct zone. RESULTS: Multi-detector computed tomography demonstrated increasing thickness of sub-endocardial viable myocardium in the infarct zone in MSC-treated animals (1.0 +/- 0.2 mm to 2.0 +/- 0.3 mm, 1 and 8 weeks after MI, respectively, p = 0.028, n = 4) and a corresponding reduction in infarct scar (5.1 +/- 0.5 mm to 3.6 +/- 0.2 mm, p = 0.044). No changes occurred in control subjects (n = 4). Tagging MRI demonstrated time-dependent recovery of active contractility paralleling new tissue appearance. This rim was composed of morphologically normal cardiomyocytes, which were smaller in MSC-treated versus control subjects (11.6 +/- 0.2 mum vs. 12.6 +/- 0.2 mum, p < 0.05). CONCLUSIONS: With serially obtained MRI and MDCT, we demonstrate in vivo reappearance of myocardial tissue in the MI zone accompanied by time-dependent restoration of contractile function. These data are consistent with a regenerative process, highlight the value of noninvasive multimodality imaging to assess the structural and functional basis for myocardial regenerative strategies, and have potential clinical applications. PMID- 17112999 TI - Imaging stem cells implanted in infarcted myocardium. AB - Stem cell-based cellular cardiomyoplasty represents a promising therapy for myocardial infarction. Noninvasive imaging techniques would allow the evaluation of survival, migration, and differentiation status of implanted stem cells in the same subject over time. This review describes methods for cell visualization using several corresponding noninvasive imaging modalities, including magnetic resonance imaging, positron emission tomography, single-photon emission computed tomography, and bioluminescent imaging. Reporter-based cell visualization is compared with direct cell labeling for short- and long-term cell tracking. PMID- 17113002 TI - Relationship between coronary artery calcification detected by electron-beam computed tomography and abnormal stress echocardiography: association and prognostic implications. AB - OBJECTIVES: The purpose of this study was to compare the results and prognostic value of electron-beam computed tomography (EBCT) and exercise echocardiography. BACKGROUND: Although patients with elevated coronary artery calcium scores (CACS) might be referred for exercise echocardiography, the association of EBCT CACS with wall motion score index (WMSI) is not known. METHODS: Patients without known coronary artery disease who underwent both clinically indicated EBCT and exercise echocardiography within a 3-month period were identified. Exercise WMSI was based on a 16-segment model (normal = 1; abnormal >1). The EBCT CACS was derived with the Agatston scoring system. Follow-up was obtained for the combined end point of death and myocardial infarction. RESULTS: The study population included 556 patients (age 54 +/- 10 years; 65% male). Correlation between EBCT CACS and exercise WMSI was limited (r = 0.17, p < 0.0001) but statistically significant. The proportion of patients with abnormal exercise WMSI increased with increasing CACS severity (chi-square = 19.1, p < 0.001). However, even in those with CACS >400, 66% had normal exercise WMSI. Age, CACS, and chest pain were independently associated with abnormal exercise WMSI. Events occurred in 12 (2%) patients. Wall motion score index (risk ratio [RR] 3.7, p = 0.023) and age (RR 1.9, p = 0.019) were associated with events. CONCLUSIONS: Electron-beam computed tomography CACS was predictive of abnormal exercise WMSI, but the majority of patients with elevated CACS had normal WMSI. Wall motion score index and age were the best predictors of events. Prospective studies are indicated to establish the relative roles of these tests in risk stratification. PMID- 17113003 TI - Cardiovascular magnetic resonance in arrhythmogenic right ventricular cardiomyopathy revisited: comparison with task force criteria and genotype. AB - OBJECTIVES: We sought to assess the utility of cardiovascular magnetic resonance (CMR) in the evaluation of arrhythmogenic right ventricular cardiomyopathy (ARVC) in relation to diagnostic criteria and genotype. BACKGROUND: Timely diagnosis of ARVC is difficult as clinical findings may be subtle and nonspecific in early disease. The role of CMR is controversial owing to the absence of a standardized protocol, insufficient experience with the modality, and inherent difficulties in imaging the right ventricle. METHODS: Comprehensive CMR examination was performed in 232 patients undergoing evaluation for suspected ARVC. CMR outcomes were compared with: 1) prospective clinical diagnosis using Task Force guidelines, with and without the proposed modifications for familial ARVC; and 2) gene carrier status in 35 individuals from genotyped families. RESULTS: CMR studies were positive in all 64 patients who prospectively fulfilled Task Force criteria, resulting in 100% sensitivity. Specificity in relation to Task Force criteria was low (29%). Of the 119 apparent false positives detected by CMR, however, 63 fulfilled modified diagnostic criteria for familial ARVC and 7 were obligate gene carriers, suggesting that CMR frequently identifies individuals with early disease, in whom Task Force criteria are relatively insensitive. This was borne out by evaluation of genotyped individuals (26 gene-positive and 9 gene negative), in whom CMR had a sensitivity of 96% and a specificity of 78%. CONCLUSIONS: CMR is a valuable component of the diagnostic workup for ARVC when performed with a dedicated protocol by specialists with experience in analysis of volumes, right ventricular wall motion, and delayed-enhancement imaging. PMID- 17113004 TI - Achieving quality in cardiovascular imaging: proceedings from the American College of Cardiology-Duke University Medical Center Think Tank on Quality in Cardiovascular Imaging. AB - Cardiovascular imaging has enjoyed both rapid technological advances and sustained growth, yet less attention has been focused on quality than in other areas of cardiovascular medicine. To address this deficit, representatives from cardiovascular imaging societies, private payers, government agencies, the medical imaging industry, and experts in quality measurement met, and this report provides an overview of the discussions. A consensus definition of quality in imaging and a convergence of opinion on quality measures across imaging modalities was achieved and are intended to be the start of a process culminating in the development, dissemination, and adoption of quality measures for all cardiovascular imaging modalities. PMID- 17113005 TI - Improving imaging: our professional imperative. AB - Many factors, including disproportionate growth rates and exciting new technologies, have focused attention on cardiovascular imaging. However, critical examination of the field reveals a surprisingly weak evidence base and inconsistent systematic attention to quality improvement. Remedies span research and practice. The optimal clinical continuum of care begins with ensuring a proper match between the diagnostic test and the individual's clinical question, and progresses to include image acquisition, image interpretation, and results reporting. Better research methodologies are needed to more tightly link imaging use to improved outcomes in non-biased community populations. To accomplish these lofty goals, alignment across stakeholders is needed to ensure the necessary human and capital investment in research and systems of care. PMID- 17113006 TI - Wither the cardiac physical examination? PMID- 17113007 TI - Chemotherapy and diagnosis of tuberculosis. AB - Since after the first streptomycin 1944 trials, anti-tuberculous chemotherapy research has been focused upon establishing drug combination regimens capable of overcoming drug resistance and amenable to ambulatory treatment in resource strapped countries. The first milestone being the 1959 Madras trial comparing home and sanatorium treatment in South India. Subsequently, the MRC trials led Fox and Mitchison to indicate rifampicin, isoniazid and pyrazinamide as the first line drugs for short course, 6 month, regimens and the 1982 Hong Kong Chest Service trials established intermittent therapy as the ambulatory treatment standard for directly observed therapy (DOT). The rising of the HIV epidemic at the beginning of the 1980s has refuelled tuberculosis spread in Africa and Asia and contributed to the expansion of drug-resistant tuberculosis worldwide making the development of new drugs and drug regimens for ambulatory treatment a top priority. Led by biotechnological advances, molecular biology has been brought into TB laboratory diagnosis for the highly sensitive and specific rapid identification of Mycobacterium tuberculosis in biological samples. The field of immunological diagnosis of TB infection, dominated since the early 1900s by the intradermal tuberculin reaction has been put back in motion by the discovery of M. tuberculosis-specific proteins and peptides, now employed in blood tests of high sensitivity and specificity for the diagnosis of latent TB which may help with the identification of contacts at higher risk of active disease and the eradication of epidemic cases. PMID- 17113008 TI - Talc poudrage versus talc slurry in the treatment of malignant pleural effusion. A prospective comparative study. AB - OBJECTIVE: The aim of this study was to investigate the effectiveness, safety and appropriate mode of administration of intrapleural talc for pleurodesis, in the treatment of malignant pleural effusion (MPE). METHODS: Prospective not randomized trial was conducted to compare thoracoscopic talc poudrage (TP) with tube thoracostomy and talc slurry (TS) for the local control of malignant pleural effusion. Both procedures were previously standardized; 6g of talc was administered for each procedure. Only the patients with lung re-expansion after drainage entered the study. Patients at high risk for general anaesthesia, poor general conditions and short life-expectancy received talc slurry through a chest tube, at the bedside. All the other patients underwent videothoracoscopic talc poudrage, with a pneumatic atomizer, under general anaesthesia. Morbidity, 30-day freedom from recurrence and long-term results were assessed and the two groups were compared. RESULTS: One hundred and nine patients entered the study (72 TP, 37 TS). Sixty-three patients in the TP group (87.5%) and 27 in the TS group (73%) had an immediate successful pleurodesis (p = 0.049); 53 patients (88.3%) and 16 patients (69.6%) had a successful pleurodesis 90 days after the procedure; 59 patients (81.9%) and 23 patients (62.2%), respectively, had a life-long pleural symphysis (p = 0.023). Adverse effects were generally mild: chest pain (36.1% in TP patients, 48.6% in TS patients) and fever (38.8% and 35.1%, respectively) were the more common but the difference was not significant between the two groups. We observed neither acute respiratory failure nor mortality due to the procedure. CONCLUSIONS: Our study confirms that intrapleural talc carries good results in the treatment of malignant pleural effusion. TP was significantly more effective than TS; both methods were safe but TS had a higher incidence of thoracic pain during the procedure. Talc pleurodesis should be offered to every patient with MPE, apart from terminally ill ones, provided that a satisfying lung re-expansion has been achieved. TP should be performed whenever possible; otherwise, a slurry bedside procedure will be worthwhile, even in patients with low performance status (PS), though poorer results have to be expected. A careful selection is essential to define the proper technique. PMID- 17113009 TI - Vascular tumors of infancy and childhood: beyond capillary hemangioma. AB - Vascular tumors of infancy and childhood represent a number of clinicopathologically distinct entities for which precise histopathological diagnosis is often essential in determining effective therapeutic approach. Unfortunately, pathologists and clinicians alike have traditionally tended to lump these tumors, in addition to small vessel vascular malformations, under overly generic terms like capillary hemangioma that do little, if anything, to guide proper clinical management. In the last decade this nosologic oversimplification has begun to wane as important new diagnostic tools and better understanding of etiology have evolved, facilitated by international recognition of the need for a multidisciplinary approach in dealing with these perplexing and often clinically devastating lesions. This article provides a brief historical perspective on this progress, and then focuses on the current clinical, histological, and immunophenotypical features that distinguish the major types of vascular tumors of infancy and childhood, also reviewing new evidence regarding their mechanisms of pathogenesis. PMID- 17113010 TI - Small animal models of cardiovascular disease: tools for the study of the roles of metabolic syndrome, dyslipidemia, and atherosclerosis. AB - INTRODUCTION: Cardiovascular disease, the leading cause of death in much of the modern world, is the common symptomatic end stage of a number of distinct diseases and, therefore, is multifactorial and polygenetic in character. The two major underlying causes are disorders of lipid metabolism and metabolic syndrome. The ability to develop preventative and ameliorative treatments will depend on animal models that mimic human disease processes. The focus of this review is to identify suitable animal models and insights into cardiovascular disease achieved to date using such models. CRITERIA FOR ANIMAL MODELS: The ideal animal model of cardiovascular disease will mimic the human subject metabolically and pathophysiologically, will be large enough to permit physiological and metabolic studies, and will develop end-stage disease comparable to those in humans. Given the complex multifactorial nature of cardiovascular disease, no one species will be suitable for all studies. AVAILABLE MODELS: Potential larger animal models are problematic due to cost, ethical considerations, or poor pathophysiological comparability to humans. Rabbits require high-cholesterol diets to develop cardiovascular disease, and there are no rabbit models of metabolic syndrome. Spontaneous mutations in rats provide several complementary models of obesity, hyperlipidemia, insulin resistance, and type 2 diabetes, one of which spontaneously develops cardiovascular disease and ischemic lesions. The mouse, like normal rats, is characteristically resistant to cardiovascular disease, although genetically altered strains respond to cholesterol feeding with atherosclerosis, but not with end-stage ischemic lesions. CONCLUSIONS: The most useful and valid species/strains for the study of cardiovascular disease appear to be small rodents, rats, and mice. This fragmented field would benefit from a consensus on well-characterized appropriate models for the study of different aspects of cardiovascular disease and a renewed emphasis on the biology of underlying diseases. PMID- 17113011 TI - Primary cardiac amyloidosis with 20-year survival. AB - BACKGROUND: The natural history of primary amyloidosis is poor, and for patients with symptomatic cardiac involvement, survival is generally less than 6 months. Even among treated patients with amyloid heart disease, survival beyond 5 years is rare. CASE REPORT: We report a patient with primary cardiac amyloidosis who is currently alive 20 years after his initial diagnosis. The extent and subtype of amyloid were documented by endomyocardial biopsy both at the time of initial diagnosis and 20 years later. To our knowledge, this is the longest survival ever reported for a patient with cardiac involvement by primary amyloidosis. CONCLUSION: The remarkably long stabilization of amyloid deposition in this patient may be attributed to early diagnosis, early institution of therapy, and, possibly, favorable genetic factors. PMID- 17113012 TI - Prebypass histological and ultrastructural evaluation of the long saphenous vein as a predictor of early graft failure. AB - BACKGROUND: Twenty percent of the long saphenous vein (LSV) grafts that are employed as coronary bypass conduits occlude during the first year after the operation. The aim of this study was to evaluate the morphological parameters of the LSV grafts before implantation as predictors for the early occlusion of the grafts. METHODS: Forty-two samples of LSV grafts were examined via light, transmission electron, and scanning electron microscopy and evaluated clinically and by angiography at 6 months and 2 years after the operation. Morphological parameters were statistically analyzed and examined for their significance on the viability of the vein grafts. RESULTS: Six (14.28%) of the examined grafts occluded within the first 6 months after the operation, and 11 grafts (26.19%) occluded within the first 2 years. The grafts that occluded at 6 months were characterized by thick intima (mean value, 206+/-32.29 vs. 67.44+/-10.17 in the group functioning normally and 98.42+/-34 in the group occluded within 2 years), low endothelial coverage (22.7+/-4.04 vs. 64.61+/-2.89 and 26.06+/-1.78 in the corresponding groups), and narrow lumen (46.73+/-9.69 vs. 527.18+/-45.78 and 204.26+/-16.5 in the corresponding groups). The presence of foam cells, edema, calcification, neovascularization, and thrombus in the lumen of the veins is frequently observed in the wall of the occluded vein grafts, whereas fibrosis does not seem to be related. CONCLUSIONS: LSV grafts with low endothelial cell coverage, stenosis of the lumen, and thick walls are at an increased risk of developing intrawall lesions that lead to early graft failure. PMID- 17113013 TI - Cardiomyopathy with a unique finding of bicuspid aortic valve in Becker's muscular dystrophy. AB - We describe a patient with Becker's muscular dystrophy and cardiac failure caused by a combination of dilated cardiomyopathy and congenital bicuspid aortic valve with aortic stenosis. There is no documented association between congenital valve disease and human dystrophinopathies, and to our knowledge, this is the first reported case. PMID- 17113014 TI - Pulmonary hypertension and patent ductus arteriosus in an 82-year-old female patient. AB - An 82-year-old female patient with a history of deep vein thrombosis presented with progressive dyspnea. Echocardiogram demonstrated significant pulmonary hypertension and patent ductus arteriosus (PDA). There was considerable debate regarding the role of PDA in the patient's pulmonary hypertension. The patient died of heart failure a few months later. Autopsy demonstrated extensive chronic pulmonary thromboembolic disease. Pulmonary thromboemboli continue to be a diagnostic challenge despite modern diagnostic modalities. Autopsy continues to play a role in investigating unexplained clinical findings and in determining cause of death. PMID- 17113015 TI - Huge biatrial cardiac rhabdomyosarcoma resulting in bilateral atrioventricular valve obstruction. AB - We present a case report of a 28-year-old man with a huge tumor that occupies both dilated atria almost completely but not extending into the ventricles. Cardiac rhabdomyosarcoma (CR) is extremely rare in adult patients. Its rarity and heterogenous clinical presentation make diagnosis difficult. In our case, after we have made an initial diagnosis via echocardiography, we took biopsies under computed tomography (CT) guiding. Histologic examination showed a CR. We want to emphasize that CT-guided biopsy is a useful and applicable technique in establishing preoperative diagnosis of intracardiac masses. PMID- 17113016 TI - Diagnosis, imaging, and treatment of an unusual cardiac hydatid cyst. AB - Echinococcosis is endemic in sheep- and cattle-raising areas in Europe, especially in Southern and Central Europe. In France, most cases originated from immigrants from countries where echinococcosis is endemic. Extremely rare native cases have been reported during the last few years in France, especially those concerning isolated cardiac hydatid cyst. In this case report, we propose a complete imaging description of the features of a typical cardiac hydatid cyst from cardiac MRI, complete with surgery, parasitology, and anatomopathology images. PMID- 17113017 TI - A recommended specification for heavy vehicle rear underrun guards. AB - A large research program was carried out to evaluate the effectiveness of different heavy vehicle rear underrun guards. Based on the test results, a recommended underrun guard specification was developed to prevent passenger injuries in such accidents. Data obtained from a series of static and full-scale crash tests performed at the Transport Canada Research Center was used in the development of the specification. Four different underrun guard designs were used during the full-scale crash tests. Three different vehicle models representing sub-compact, compact vehicles and light trucks and vans traveling at 48, 56 and 65 km/h relative speeds were used to impact underrun guards head on. The first 10 of the crash tests guard was attached to a simulated trailer while the last one performed using an actual truck. The results obtained from these tests show that a guard built to the minimum requirements of U.S. FMVSS 223 may not be strong enough to prevent passenger compartment intrusion especially in compact and sub compact automobiles. This paper documents the tests performed, the results obtained and their analyses, and conclusions derived from these analyses. A recommended specification is prepared on the minimum performance that should be required for an effective underrun guard. PMID- 17113018 TI - Estimating the relationship between measured wind speed and overturning truck crashes using a binary logit model. AB - This paper develops a quantitative model that correlates overturning freight vehicle crash records in Wyoming to measured wind speeds at nearby weather stations. The database consists of 14,700 truck crashes from 1994 to 2003 and wind speed and gust information from 21 weather stations. A binary logit model was estimated from the data to determine if there was significant correlation between weather station wind data and the likelihood that the crash was of the overturning type. While it is reasonably known that local wind speeds at the crash location are critical in predicting overturning truck crash likelihood, it was not known if distant weather station data were an adequate predictor of these crash types. The results from this work indicate that weather station data can be used as a predictor of overturning crashes. This work provides the necessary first step for the development of operational rules for roadway sections that run high risk of overturning truck crashes in high wind conditions. PMID- 17113019 TI - Intentional action arises from early reciprocal exchanges. AB - Intentionality is defined as the cognitive ability to represent goals beyond the here and now of perception. First signs of intentionality appear by the second month after birth. A major mechanism responsible for such development might be the unique reciprocal and intentional ways humans communicate with each other, particularly their young progenies. I argue that starting in the second month, reciprocal exchanges, affective mirroring, and mutual imitations with others provide infants with the unique opportunity to differentiate as well as to compare and conjugate first (self) and third (others') person perspectives. This ability is a prerequisite not only for referential communication but also of intentionality. It corresponds to the emergence of a new contemplative and "meta" stance toward the world. An argument is made that the developmental origins of this stance are primarily social, not to be found in the individual infant interacting solely with physical objects. PMID- 17113020 TI - Developmental changes in oculomotor control and working-memory efficiency. AB - In the present study, we examined the developmental changes in the efficiency of saccadic inhibitory control. More specifically, the contribution of age-related changes in working-memory engagement was investigated. We manipulated the efficiency of inhibitory oculomotor control in antisaccade tasks by using fixation-offset conditions, which are supposed to affect inhibitory demands, and by adding increasing working-memory loads to the antisaccade task. In general, in comparison to antisaccade performance of adults, the antisaccade performance of 8 year-old and 12-year-old children was characterized by an increase in direction errors, and/or longer saccadic onset latencies on correct antisaccades. However, this pattern was not altered by the fixation-offset manipulations. In contrast, increased working-memory demands deteriorated 8-year-olds' antisaccade performance unequally as compared to older children and young adults. These findings suggest that - at least in young children - the available functional working-memory capacity is engaged in oculomotor inhibition. PMID- 17113021 TI - Antibody responses to Plasmodium falciparum vaccine candidate antigens in three areas distinct with respect to altitude. AB - Antibody levels against malaria antigens were measured among patients presenting with uncomplicated malaria at health centers from three locations in Zimbabwe (Bindura, Chiredzi and Kariba) that are distinct with regard to altitude and climatic conditions. Antibody levels were determined by ELISA using the antigens, apical membrane antigen 1 (AMA-1), erythrocyte binding antigen 175 (EBA-175), circumsporozoite surface protein (CSP), merozoite surface protein 1 (MSP-1) and Pfg27. For all the antigens tested, IgG and IgM levels were higher for Bindura (altitude 1100 m) compared to Kariba (<600 m, altitude) and Chiredzi (approximately 600 m, altitude) with the exception of IgG and IgM to AMA-1 and EBA-175 which were similar between Chiredzi and Bindura. Plasma samples were further analyzed for their functional activity by testing their ability to inhibit the growth of Plasmodium falciparum in culture. Our results, determined by microscopy and verified by the LDH assay revealed that plasma from the three locations had similar inhibitory activity against the growth of P. falciparum in vitro. Our data revealed that highest growth inhibition correlated with the highest levels of MSP-1 antibody values. PMID- 17113022 TI - Real-time PCR: duplexing without optimization. AB - Real-time PCR applications generally require determination of the threshold cycle of a gene of interest in parallel with an endogenous control. In standard situations, the gene-specific and control reactions are run as separate samples (singleplex). In contrast, duplex approaches combine both reactions within a single well, thereby saving time, cost, and material. However, establishing duplex reactions usually requires laborious optimization justifiable only for the analysis of large sample series. Hence, in research settings, singleplex approaches are used most commonly. To establish conditions for duplexing without the need of optimization, we tested the performance of 40 premade TaqMan gene expression assays in duplex reactions with an endogenous control using three different polymerase mixes. The results were compared with singleplex reactions. Duplexing results obtained with one of the multiplex polymerase mixes correlated extremely well (r(2)=0.95) with the singleplex reference. The findings of our study demonstrate that the combination of this polymerase mix and premade gene expression assays will yield reliable and reproducible results in duplex approaches without preceding optimization. PMID- 17113023 TI - Direct quantification of gene expression using capillary electrophoresis with laser-induced fluorescence. AB - Quantification of gene expression provides valuable information regarding the response of cells or tissue to stimuli and often is accomplished by monitoring the level of messenger RNA (mRNA) being transcribed for a particular protein. Although numerous methods are commonly used to monitor gene expression, including Northern blotting, real-time polymerase chain reaction, and RNase protection assay, each method has its own drawbacks and limitations. Capillary electrophoresis with laser-induced fluorescence (CE-LIF) can reduce protocol time, eliminate the need for radioactivity, and provide superior sensitivity and dynamic range for quantification of RNA. In addition, CE-LIF can be used to directly determine the amount of an RNA species present, something that is difficult and not normally accomplished using current methods. Gene expression is detected using a fluorescently labeled riboprobe specific for a given RNA species. This direct approach was validated by analyzing levels of 28S RNA and also used to determine the amount of discoidin domain receptor 2 mRNA in cardiac tissue. PMID- 17113024 TI - Accurate identification of desired clones from 16S-23S rDNA spacer libraries using single PCR. PMID- 17113025 TI - Intercalators: contra cruciform extrusion in DNA. AB - Although there is a wealth of structural and theoretical data relating to palindromic sequences in genomes, the mechanisms of extrusion of cruciform structures during various biological processes in the presence of intercalating agents are still poorly understood. The current study addresses the effects of temperature and intercalator on cruciform extrusion from plasmids and also considers the effects of divalent metal ions on cruciform extrusion. It presents evidence that the cytotoxic effects of certain DNA binding drugs in vivo occur over concentration ranges corresponding to those that modulate cruciform extrusion in vitro. The results confirm earlier studies showing an inverse relationship between the effects of negative superhelicity and temperature on cruciform extrusion. By extrapolation, divalent metal ions facilitate cruciform extrusion by increasing superhelicity. The results allow the concentrations that preclude cruciform extrusion in DNA to be determined, and these are potentially informative about the relationships among temperature, DNA helical winding, cruciform formation, and intercalation. Overall, we provide new and interesting insights into the potential role of cruciform structures in biology and, by implication, cancer therapy. PMID- 17113026 TI - An HPLC-based fluorometric assay for cobalamin-independent methionine synthase. PMID- 17113027 TI - Construction of an XcmI-generated T vector bearing green fluorescent protein marker for direct cloning of PCR products. PMID- 17113028 TI - Fluorescent labeling of cell-free synthesized proteins by incorporation of fluorophore-conjugated nonnatural amino acids. AB - Although fluorescent dyes, such as fluorescein derivatives, have bulky and complex structures, nonnatural amino acids carrying these fluorescein derivatives are acceptable by the Escherichia coli ribosome and are useful for the cotranslational fluorescent labeling of cell-free synthesized proteins. Surprisingly, the incorporation efficiency of nonnatural amino acids carrying fluorescein derivatives into translated proteins depends on the source of the translational machinery used in cell-free protein synthesis. That is, whereas the E. coli ribosome efficiently supported the incorporation of nonnatural amino acids carrying fluorescein derivatives into a protein structure, no detectable fluorescent signal was observed from the protein expressed in the eukaryotic cell free protein synthesis system performed in the presence of fluorescein-conjugated aminoacylated transfer RNA (tRNA). PMID- 17113029 TI - Homemade viral RNA isolation protocol using silica columns: a comparison of four protocols. PMID- 17113030 TI - A thin-layer electrophoretic assay for Asp-tRNAAsn/Glu-tRNAGln amidotransferase. PMID- 17113031 TI - The effects of cycle time on the physical demands of a repetitive pick-and-place task. AB - This study seeks to elucidate the effects of the cycle time of a pick-and-place task on muscle activity, grip force, posture, and perception-based measures (discomfort and difficulty). Six healthy adults (3 males, 3 females) participated. A 4 x 2 repeated measures design was used with cycle time (1, 2, 5, and 10s) and grip (power and chuck) as independent variables. The task consists of repetitively picking a 0.7 kg part and placing it into a bin. A reduction in cycle time (CT) resulted in both a decrease of task time and physical rest time (p<0.001). The physiological muscle rest was much lower than the physical rest time (p<0.05). An increase in static muscle loading (p<0.01), grip force (p<0.001), and discomfort (p<0.001) were also observed. These results suggest that a pace threshold (between 2 and 5s for this task) is reached at a higher CT than that defined by the ability to perform the task. PMID- 17113032 TI - Rho, Rho-kinase, and the actin cytoskeleton regulate the Na+ -H+ exchanger in sea urchin eggs. AB - At fertilization, the sea urchin egg undergoes an internal pH (pHi) increase mediated by a Na+ -H+ exchanger. We used antibodies against the mammalian antiporters NHE1 and NHE3 to characterize this exchanger. In unfertilized eggs, only anti-NHE3 cross-reacted specifically with a protein of 81-kDa, which localized to the plasma membrane and cortical granules. Cytochalasin D, C3 exotoxin (blocker of RhoGTPase function), and Y-27632 (inhibitor of Rho-kinase) prevented the pHi change in fertilized eggs. These inhibitors blocked the first cleavage division of the embryo, but not the cortical granule exocytosis. Thus, the sea urchin egg has an epithelial NHE3-like Na+ -H+ exchanger which can be responsible for the pHi change at fertilization. Determinants of this pHi change can be: (i) the increase of exchangers in the plasma membrane (via cortical granule exocytosis) and (ii) Rho, Rho-kinase, and optimal organization of the actin cytoskeleton as regulators, among others, of the intrinsic activity of the exchanger. PMID- 17113033 TI - Discoidin domain receptor 2 is involved in the activation of bone marrow-derived dendritic cells caused by type I collagen. AB - Discoidin domain receptors (DDRs), DDR1 and DDR2, are non-integrin receptor tyrosine kinases for collagen in many cell types. In this study, we investigated the contributions of DDRs to the activation of mouse bone marrow-derived dendritic cells (DCs) by type I collagen (ColI). Our data showed that transcript and protein of DDR2 were expressed constitutively in immature DCs and upregulated in TNF-alpha-stimulated mature DCs. ColI treatment induced DDR2 phosphorylation and subsequently induced the upregulation of IL-12 production, CD86 expression, and antigen uptake activity by immature DCs. Depletion of DDR2 by specific siRNA attenuated significantly an increase in expression of IL-12 and CD86 in ColI treated DCs. Additionally, DDR2-ColI interaction upregulated the ability of mature DCs to activate allogeneic T cells. These findings suggest that DDR2 is a critical collagen receptor for DC activation and that DDR2-collagen interaction plays an important role in the functional capacity of DCs regulating immune responses. PMID- 17113034 TI - Dicer1 expression in preimplantation mouse embryos: Involvement of Oct3/4 transcription at the blastocyst stage. AB - Dicer1, an RNAse III enzyme, is a key factor for the production of microRNAs involved in post-transcriptional gene silencing. To elucidate the roles of Dicer1 and the microRNA pathway in early embryo development, we initially evaluated its gene expression in mouse oocytes and embryos in vitro. The transcript levels in GV stage oocytes steadily decreased up to the 2-cell embryo stage, and expression remained stable during morulae and blastocyst formation. DICER1 protein synthesis was additionally observed in mouse oocytes and early embryos. Silencing of mRNA expression by RNA interference (siRNA) did not inhibit development up to the blastocyst stage. Real-time RT-PCR experiments confirmed the decreased expression of selected transcription factors, including POU domain, class 5, transcription factor 1 (Pou5f1), SRY-box containing gene 2 (Sox2), and Nanog homeobox (Nanog). Moreover, POU5F1 protein expression was suppressed by Dicer1 siRNA. The results suggest that Dicer1 gene expression is associated with the levels of transcription factors, Pou5f1, Sox2, and Nanog which possibly regulate differentiation processes at the blastocyst stage. PMID- 17113035 TI - Solid-phase classical complement activation by C-reactive protein (CRP) is inhibited by fluid-phase CRP-C1q interaction. AB - C-reactive protein (CRP) interacts with phosphorylcholine (PC), Fcgamma receptors, complement factor C1q and cell nuclear constituents, yet its biological roles are insufficiently understood. The aim was to characterize CRP induced complement activation by ellipsometry. PC conjugated with keyhole limpet hemocyanin (PC-KLH) was immobilized to cross-linked fibrinogen. A low-CRP serum with different amounts of added CRP was exposed to the PC-surfaces. The total serum protein deposition was quantified and deposition of IgG, C1q, C3c, C4, factor H, and CRP detected with polyclonal antibodies. The binding of serum CRP to PC-KLH dose-dependently triggered activation of the classical pathway. Unexpectedly, the activation was efficiently down-regulated at CRP levels > 150 mg/L. Using radial immunodiffusion, CRP-C1q interaction was observed in serum samples with high CRP concentrations. We propose that the underlying mechanism depends on fluid-phase interaction between C1q and CRP. This might constitute another level of complement regulation, which has implications for systemic lupus erythematosus where CRP is often low despite flare-ups. PMID- 17113036 TI - PRIMA-1 induces apoptosis by inhibiting JNK signaling but promoting the activation of Bax. AB - The p53 protein plays a major role in the maintenance of genome stability in mammalian cells. Mutations of p53 occur in over 40% of breast cancers and are indicative of tumor resistance to chemotherapeutic agents. Recently, there has been a high degree of interest in pharmacological approaches for restoring the normal function to mutant p53. The low molecular weight compound p53 reactivation and induction of massive apoptosis (PRIMA-1) was shown to induce cytotoxic effects and apoptosis in human tumor cells with mutant p53. Here, we studied the molecular mechanisms of PRIMA-1-induced apoptosis in human breast cancer cells with p53 mutations such as MDA-231 and GI-101A as compared to MCF-7 cells. We show that PRIMA-1 selectively induces apoptosis in human breast cancer cells MDA 231 and GI-101A compared to the MCF-7. This effect was paralleled by an increase in total p53 level in the nucleus and the induction of its phosphorylation at Ser 15 site. Using the chromatin immunoprecipitation (ChIP) assays, we show that PRIMA-1 restored p53 DNA binding activity to the promoters of the proapoptotic genes such as Bax and PUMA, but inhibited the binding activity to the promoters of the MAP4K4 gene. Knockdown of p53 protein in breast cancer cells using siRNA followed by PRIMA-1 treatment resulted in decline of Bax and PUMA proteins expression. Cell incubation with either PRIMA-1 or SP600125 (c-Jun NH2-terminal kinase inhibitor) resulted in the abrogation of adriamycin-induced c-Jun NH2 terminal kinase (JNK) activation, whereas Bax activation was not inhibited. We conclude that both Bax and PUMA but not JNK signaling are involved in PRIMA-1 induced apoptosis in breast cancer cells with p53 mutation. PMID- 17113037 TI - Efficient protein selection based on ribosome display system with purified components. AB - Using the PURE (Protein synthesis Using Recombinant Elements) system, we developed an efficient and highly controllable ribosome display method for selection of functional protein. The PURE system is composed of purified factors and enzymes that are responsible for gene expression in Escherichia coli. We performed the detailed analyses and optimization of the ribosome display system and demonstrated the formation of stable mRNA/ribosome/polypeptide ternary complexes. As complex formation is fundamental to successful ribosome display, these improvements resulted in a dramatic increase in the mRNA recovery rate. As a result, a approximately 12,000-fold enrichment of single-chain antibody (scFv) cDNA was achieved in a single round of selection. Specific selection of scFv mRNA from a 1:10(10) dilution in competitor mRNA was achieved with only three rounds of affinity selection. These findings, together with the results in the accompanying paper [T. Matsuura, H. Yanagida, J. Ushioda, I. Urabe, T. Yomo, Nascent chain, RNA, and ribosome complexes generated by pure translation system (see the accompanying paper).], demonstrate that the PURE system can provide a basis for reliable and reproducible ribosome display. PMID- 17113038 TI - The CSPalpha/G protein complex in PC12 cells. AB - Cysteine string proteinalpha (CSPalpha) is a regulated vesicle protein and molecular chaperone that has been found to be critical for continuous synaptic transmission and is implicated in the defense against neurodegeneration. Previous work has revealed links between CSPalpha and heterotrimeric GTP binding protein (G protein) signal transduction pathways. We have shown that CSPalpha is a guanine nucleotide exchange factor (GEF) for Galphas. In vitro Hsc70 (70 kDa heat shock cognate protein) and SGT (small glutamine-rich tetratricopeptide repeat domain protein) switch CSPalpha from an inactive GEF to an active GEF. Here we have examined the cellular distribution of the CSPalpha system in the PC12 neuroendocrine cell line. CSPalpha, an established secretory vesicle protein, was found to concentrate in the processes of NGF-differentiated PC12 cells as expected. Gbeta subunits co-localized and Galphas subunits partially co-localized with CSPalpha. However, under the conditions examined, the GEF activity of CSPalpha is expected to be inactive, in that Hsc70 was not found in PC12 processes. These results indicate that CSPalpha activity is subject to regulation by factors that alter Hsc70 distribution and translocation within the cell. PMID- 17113039 TI - Aurora kinase inhibition downregulates NF-kappaB and sensitises tumour cells to chemotherapeutic agents. AB - We have identified that Aurora-A activates NF-kappaB via IkappaBalpha phosphorylation. Here, we analysed different human tumour cell types for their NF kappaB activity. We found that there is an association between cell resistance to chemotherapeutic agents and NF-kappaB activation. A549 human lung adenocarcinoma cells and SKOV3 human ovarian cancer cells have high levels of NF-kappaB and are resistant to cytotoxic agents such as adriamycin and VP-16 (etoposide). We also found that in A549 and SKOV3 cells treated with a small molecule inhibitor towards Aurora kinases, the NF-kappaB activity was downregulated and the efficacy of cytotoxic drugs was enhanced. In addition, the transcriptional targets Bcl-XL and Bcl-2 were downregulated. This study provides evidence for a potential mechanism of chemoresistance and may be useful for the enhancement of certain chemotherapeutics regimens. PMID- 17113040 TI - Inhibition of nitric oxide-guanylate cyclase-dependent and -independent signaling contributes to impairment of beta-adrenergic vasorelaxations by cyclosporine. AB - This study investigated the role of endothelium- and smooth muscle-dependent mechanisms in the interaction of cyclosporine (CyA), an immunosuppressant drug, with beta-adrenoceptor (isoprenaline)-mediated relaxations in isolated rat aortas precontracted with phenylephrine. CyA effects were assessed in the absence and presence of NG-nitro-L-arginine methyl ester (L-NAME, nitric oxide synthase inhibitor), methylene blue (guanylate cyclase inhibitor), or propranolol (beta adrenoceptor antagonist). In aortas with intact endothelium (E+), pretreatment with L-NAME or methylene blue significantly reduced isoprenaline (1 x 10(-9) to 1 x 10(-7)M) relaxations in contrast to no effect for tetraethylammonium (K+ channel blocker), or diclophenac (cyclooxygenase inhibitor), suggesting a major role for the nitric oxide-guanylate cyclase (NO-GC) pathway, but not endothelial hyperpolarizing factor or vasodilator prostanoids, in isoprenaline responses. Isoprenaline relaxations were still evident, though significantly attenuated, in endothelium-denuded aortas (E-) and were resistant to L-NAME or methylene blue. Acute exposure to CyA (2 microM) caused propranolol-sensitive reductions in isoprenaline responses in E+ and E- aortas. The CyA-induced attenuation of isoprenaline responses in E+ aortas largely disappeared in L-NAME-treated aortas and after supplementation with L-arginine, the substrate of nitric oxide. CyA also reduced the endothelium-independent, GC-dependent aortic relaxations evoked by sodium nitroprusside, an effect that was virtually abolished by methylene blue. We conclude that: (i) endothelial and smooth muscle mechanisms contribute to aortic beta-adrenoceptor relaxations and both components are negatively influenced by CyA, and (ii) NO-GC signaling plays an integral role in the vascular CyA-beta-adrenoceptor interaction. The clinical relevance of the present study is warranted given the established role of impaired vascular function in CyA toxicity. PMID- 17113041 TI - Effects of leptin on apoptosis and adipogenesis in 3T3-L1 adipocytes. AB - Leptin has been demonstrated to induce adipose tissue apoptosis, which can contribute to the decrease of adiposity, after either central nervous system or peripheral administration. However, it is not known whether leptin acts only centrally to initiate a signal or can also act directly on adipocytes to induce apoptosis. The objective of this study was to determine the direct effect of leptin on adipocyte apoptosis and adipogenesis in vitro using 3T3-L1 cell lines. An ELISA for single stranded DNA, which is highly specific for apoptotic cells, was used to quantify apoptosis. Preconfluent preadipocytes treated with 10(-9), 10(-8), 10(-7), and 10(-6)M leptin showed inhibitory effects on cell viability, and similar observations were also found in maturing preadipocytes treated during day 0-2 and day 2-4 of maturation. After 48 h incubation with 10(-6)M leptin, LDH release was increased by 24.3% (p<0.05) in preconfluent preadipocytes and by 108.5% (p<0.01) in maturing preadipocytes. However, ssDNA analysis revealed no increased apoptosis in preconfluent or maturing preadipocytes or in mature adipocytes treated with leptin. Leptin significantly reduced lipid accumulation and GPDH activity in maturing preadipocytes, demonstrating an inhibitory effect of leptin on adipogenesis. These results indicate that leptin does not act directly to induce adipocyte apoptosis, but can act directly to inhibit maturation of preadipocytes. PMID- 17113042 TI - Myricetin induces human osteoblast differentiation through bone morphogenetic protein-2/p38 mitogen-activated protein kinase pathway. AB - Myricetin (3,3',4',5,5',7-hexahydroxyflavone), a flavonoid compound, is present in vegetables and fruits. By means of alkaline phosphatase (ALP) activity, osteocalcin, and type I collagen enzyme-linked immunosorbent assay (ELISA), we have shown that myricetin exhibits a significant induction of differentiation in MG-63 and hFOB human osteoblasts. Alkaline phosphatase and osteocalcin are phenotypic markers for early-stage differentiated osteoblasts and terminally differentiated osteoblasts, respectively. Our results indicate that myricetin stimulates osteoblast differentiation at various stages, from maturation to terminally differentiated osteoblasts. Induction of differentiation by myricetin is associated with increased bone morphogenetic protein-2 (BMP-2) production. The BMP-2 antagonist noggin blocked myricetin-mediated ALP activity and osteocalcin secretion enhancement, indicating that BMP-2 production is required in myricetin mediated osteoblast maturation and differentiation. Induction of differentiation by myricetin is associated with increased activation of SMAD1/5/8 and p38 mitogen activated protein kinases. Cotreatment of p38 inhibitor SB203580 inhibited myricetin-mediated ALP upregulation and osteocalcin production. In conclusion, myricetin increased BMP-2 synthesis, and subsequently activated SMAD1/5/8 and p38 MAPK, and this effect may contribute to its action on the induction of osteoblast maturation and differentiation, followed by an increase of bone mass. PMID- 17113043 TI - Cannabinoid receptors are localized to noradrenergic axon terminals in the rat frontal cortex. AB - Cannabinoid agonists exert complex actions on modulatory neurotransmitters involved in attention and cognition. Previous studies have demonstrated that acute systemic administration of the synthetic cannabinoid agonist, WIN 55,212-2, increases norepinephrine efflux in the rat frontal cortex. In an effort to elucidate whether cannabinoid (CB1) receptors are positioned to presynaptically modulate norepinephrine release in the frontal cortex, immunocytochemical detection of the CB1 receptor and the catecholamine-synthesizing enzyme dopamine beta-hydroxylase (DbetaH) was performed using confocal immunofluorescence microscopy and immunoelectron microscopy in rat brain. Fluorescence microscopy analysis of dually labeled tissue sections from the frontal cortex indicated that individual axonal processes exhibited both CB1 receptor and DbetaH immunoreactivities. Ultrastructural analysis confirmed that one-third of axon terminals containing CB1 immunolabeling also exhibited DbetaH labeling. Cortical neurons were also found to be targeted by separately labeled CB1- and DbetaH containing axon terminals. In conclusion, the present neuroanatomical data suggest that cortical norepinephrine release may be modulated, in part, by CB1 receptors that are presynaptically distributed on noradrenergic axon terminals. PMID- 17113044 TI - Circadian rhythms of TRH-like peptide levels in rat brain. AB - This is the first report of diurnal variations in the levels of thyrotropin releasing hormone-like peptides (pGlu-X-Pro-NH(2), where "X" can be any amino acid residue) in brain regions involved in mood regulation. These peptides have neuroprotective and antidepressant-like properties that may help stabilize chronobiologic systems that are often abnormal in neuropsychiatric disease. We hypothesized that diurnal fluctuations in the levels of these neuropeptides are components of the chronobiologic regulation of autonomic, behavioral and emotional states. Optimal use of these potentially therapeutic agents will benefit from an understanding of their response to, and effect on, normal vegetative, activity and sleep patterns, and the corresponding disordered patterns of mental illness. For these reasons, 16 male, 200 g, Sprague-Dawley rats were maintained for 4 weeks in a stable 12 h lights on, 12 h lights off photoperiod. Levels of TRH and TRH-like peptides were measured at 3.0 h, 10.5 h, 13.5 h and 21.0 h, where the subjective midnight was 0.0 h, by a combination of HPLC and RIA. Highly significant changes in TRH-like peptide levels were observed in the striatum, posterior cingulate, cerebellum, pyriform cortex, nucleus accumbens and medulla oblongata. TRH-like peptide levels, in general, were highly correlated with changes in TRH concentration, within and between brain regions, and may be colocalized in large glutamatergic neurons innervating the rat limbic system. We conclude that TRH-like peptides may be important components of chronobiologic systems involved in maintaining autonomic, behavioral and mood equilibria. PMID- 17113045 TI - Glutamate-related gene expression changes with age in the mouse auditory midbrain. AB - Glutamate is the main excitatory neurotransmitter in both the peripheral and central auditory systems. Changes of glutamate and glutamate-related genes with age may be an important factor in the pathogenesis of age-related hearing loss presbycusis. In this study, changes in glutamate-related mRNA gene expression in the CBA mouse inferior colliculus with age and hearing loss were examined and correlations were sought between these changes and functional hearing measures, such as the auditory brainstem response (ABR) and distortion product otoacoustic emissions (DPOAEs). Gene expression of 68 glutamate-related genes was investigated using both genechip microarray and real-time PCR (qPCR) molecular techniques for four different age/hearing loss CBA mouse subject groups. Two genes showed consistent differences between groups for both the genechip and qPCR. Pyrroline-5-carboxylate synthetase enzyme (Pycs) showed down-regulation with age and a high-affinity glutamate transporter (Slc1a3) showed up-regulation with age and hearing loss. Since Pycs plays a role in converting glutamate to proline, its deficiency in old age may lead to both glutamate increases and proline deficiencies in the auditory midbrain, playing a role in the subsequent inducement of glutamate toxicity and loss of proline neuroprotective effects. The up-regulation of Slc1a3 gene expression may reflect a cellular compensatory mechanism to protect against age-related glutamate or calcium excitoxicity. PMID- 17113046 TI - Cellular and behavioral effects of 5-HT1A receptor agonist 8-OH-DPAT in a rat model of levodopa-induced motor complications. AB - 5-HT1A autoreceptor stimulation can act to attenuate supraphysiological swings in extracellular dopamine levels following long-term levodopa treatment and may be useful in the treatment and prevention of the motor complications. The purpose of this study was to investigate cellular and behavioral effects of 5-HT1A receptor agonist 8-OH-DPAT in a rat model of levodopa-induced motor complications. Two sets of experiments were performed. First, animals were treated with levodopa (50 mg/kg with benserazide 12.5 mg/kg, twice daily), intraperitoneally (i.p.) for 22 days. On day 23, animals received either 8-OH-DPAT (1 mg/kg, i.p.) or 8-OH-DPAT plus WAY-100635 (0.1 mg/kg, i.p) or vehicle with each levodopa dose. In the second set, animals were treated either with levodopa (50 mg/kg, i.p.) plus 8-OH DPAT (1 mg/kg, i.p.) or levodopa (50 mg/kg, i.p.) plus vehicle, administered twice daily for 22 consecutive days. Our study showed that 8-OH-DPAT plus levodopa both prolonged the duration of the motor response and reduced peak turning. 8-OH-DPAT plus levodopa also decreased the frequency of failures to levodopa. Co-administration of WAY-100635, a 5-HT1A receptor antagonist, with 8 OH-DPAT eliminated the effect of 8-OH-DPAT on motor complications indicating that the observed 8-OH-DPAT responses were probably mediated at the 5-HT1A autoreceptor. Moreover, 8-OH-DPAT plus levodopa significantly reduced hyperphosphorylation of GluR1 at serine 845, which was closely associated with levodopa-induced motor complications. PMID- 17113047 TI - Modifications of the retina neuronal populations of the heterozygous mutant small eye mouse, the Sey(Dey). AB - We analyzed the modifications of the retinal neurons in a heterozygous mutant small eye mouse, the Sey(Dey). This mouse presents a mutation in chromosome 2 which affects the gene Pax6 and other nearby genes, such as the Wt1 gene and the gene of the Reticulocalbin. The eyes of these animals do not have lenses and their retinas present important morphological alterations: in the anterior portion they are joined to the cornea, they are found detached from the pigment epithelium, they present folds that form rosettes in some zones and alteration of the lamination can be observed. The partial loss of the genes affected does not prevent the formation of the different layers of the retina, but does affect its thickness, principally of the plexiform layers; moreover, the internal limiting membrane is found disorganized. All the neuronal populations are present in the retina of these animals and express the same neurochemical markers as the control animals, but the number of Pax6(+) cells is notably reduced. In these retinas a marked disorganization of the distribution of the dendrites and axons is observed and a notable reduction in the axons of ganglion cells. These results suggest that, although it does not appear determinant in the differentiation of the distinct neuronal types of the retina, the partial lack of genes of the heterozygotes +/Sey(Dey) provokes important morphological and neurochemical modifications in the cytoarchitecture of the retina. PMID- 17113048 TI - Distribution of ghrelin-immunoreactive neuronal networks in the human hypothalamus. AB - Ghrelin has been discovered as the endogenous ligand of the growth hormone secretagogue receptor (GHS-R). It stimulates growth hormone secretion and also potently increases food intake. To date, ghrelin is the only known peripheral orexigenic hormone. Recent studies have demonstrated that in addition to peripheral organs, ghrelin is also synthesized in the hypothalamus. In the present study, we examined the distribution of the ghrelin-immunoreactive (IR) elements in the human hypothalamus. Ghrelin-IR fibers were widely distributed throughout the hypothalamus. Based on the thickness of fibers, major subtypes of ghrelin-IR axons were observed: thick fibers with large varicosities and very fine axons with or without small varicosities. Dense networks of ghrelin-IR axons were observed in the hypothalamic suprachiasmatic, paraventricular, supraoptic, dorsomedial, ventromedial and infundibular nuclei and in the periventricular area. Ghrelin-IR axons also appeared in the external layer of the pituitary stalk. Ghrelin-IR cell bodies were not detected. Since hypothalamic regions innervated by ghrelin-IR axons also take part in the regulation of food intake and energy balance, the centrally synthesized ghrelin may play a major role in the central regulation of energy metabolism in humans. PMID- 17113049 TI - Volume of left amygdala subregion predicted temperamental trait of harm avoidance in female young subjects. A voxel-based morphometry study. AB - We investigated the relationship between temperamental predisposition and brain structure by using a standard questionnaire and high-resolution T1-weighted magnetic resonance image (MRI) in normal young volunteers. Fifty-six subjects completed the Japanese version of the Temperament and Character Inventory (TCI, 125 items) and underwent an MRI acquisition of the brain. The gray matter (GM) was extracted from the whole brain image of the subjects and normalized to the standard brain template using statistical parametric mapping and the optimized voxel-based morphometry (VBM) method. When the score on the harm avoidance (HA) subscale was used as a dependent variable, the multiple regression analysis revealed that the HA score positively correlated with the volume of the part of left amygdala. The region-of-interest analysis showed that the correlation was significant in the female subjects but not in the male subjects. The correlation was significant even after the effects of age, depression score, and total GM volume were taken into account. The differential correlation between the sexes may be caused by differences in hormonal condition and the vulnerability of women to socio-psychological stress. In addition, the novelty seeking (NS) score positively correlated with the GM of the left middle frontal gyrus. The volume of the tail of the right caudate nucleus positively correlated with the reward dependence (RD) score. With regard to the NS and RD scores, no significant sex difference was observed in the correlation. These results indicate that the temperamental traits measured using the questionnaire may have a morphological basis in the human brain. PMID- 17113050 TI - Effects of age and caloric intake on glutathione redox state in different brain regions of C57BL/6 and DBA/2 mice. AB - The main purpose of the present study was to determine whether specific regions of the mouse brain exhibit different age-related changes in oxidative stress, as indicated by glutathione redox state and the level of protein-glutathionyl mixed disulfides. Comparison of 3- and 21-month-old mice indicated an age-related decrease in the ratio of reduced to oxidized glutathione (GSH/GSSG) as well as a pro-oxidizing shift in the calculated redox potential (ranging from 6 to 15 mV) in the cortex, hippocampus, striatum and cerebellum, whereas there was little change in the brainstem. This pro-oxidizing shift in redox state was due to a modest decrease in GSH content occurring in all the brain regions examined, and elevations in GSSG amount that were most pronounced in the striatum and cerebellum. The regional changes in glutathione redox state were paralleled by increases in the amounts of protein-mixed disulfides. A reduction of caloric intake by 40% for a short period (7 weeks), implemented in relatively old mice (17 months), increased the GSH/GSSG ratio and redox potential at 19 months in the same brain regions that exhibited age-related decreases. The effects of age and caloric restriction were qualitatively similar in C57BL/6 and DBA/2 mice. However, young DBA/2 mice, which do not show extension of life span in response to long-term caloric restriction, had lower GSH/GSSG ratios and higher protein mixed disulfides than age-matched C57BL/6 mice. The current findings demonstrate that oxidative stress, as reflected by glutathione redox state, increases in the aging brain in regions linked to age-associated losses of function and neurodegenerative diseases. PMID- 17113051 TI - Neuronal nitric oxide synthase is upregulated in a subset of primary sensory afferents after nerve injury which are necessary for analgesia from alpha2 adrenoceptor stimulation. AB - alpha2-Adrenoceptor (AR) agonists increase in analgesic potency and efficacy after peripheral nerve injury, and their effects are blocked by neuronal nitric oxide synthase (nNOS) inhibitors and M4 muscarinic receptor antagonists only after injury. We tested whether nNOS and M4 muscarinic receptors are co-expressed in the spinal cord, and whether destruction of a subset of sensory afferents which are essential to alpha2-AR analgesia would also destroy nNOS and M4 receptor expression. Male Sprague-Dawley rats underwent left L5 and L6 spinal nerve ligation. Lumbar spinal cord was removed and immunostained for M4 muscarinic receptors and nNOS alone and for co-expression. Others received intrathecal injection of saporin linked to an antibody to the neurotrophin receptor p75(NTR), which eliminates cells expressing this receptor and the analgesic effects of alpha2-AR agonists. nNOS staining of fibers in the superficial dorsal horn was dramatically increased after spinal nerve ligation, and this was abolished by saporin linked anti-p75(NTR) treatment. In contrast, nNOS staining in dorsal horn neurons was unaltered by these manipulations. M4 receptors were present on neurons in the dorsal horn, some of which co-expressed nNOS, but their pattern of expression was not altered by these manipulations. Peripheral nerve injury increases nNOS expression in fibers in the superficial dorsal horn, some of which likely express p75(NTR), and alpha2-AR agonists may reduce injury-induced sensitization by activation of nNOS in these fibers In contrast, changes in nNOS and M4 receptor location on spinal cord neurons are not responsible for increased analgesic potency of alpha2-AR agonists after nerve injury. PMID- 17113052 TI - The neurotensin receptor agonist NT69L suppresses sucrose-reinforced operant behavior in the rat. AB - NT69L is a neurotensin analog that can be administered peripherally. It blocks amphetamine- and cocaine-induced hyperactivity in rats. It also blocks nicotine induced locomotor activity and has shown sustained efficacy in a rat model of nicotine-induced sensitization. The present study tested the effect of NT69L on responding for sucrose reinforcement on a continuous reinforcement schedule (CRF) and incrementing (FR1-FR5) discrimination schedule. Male Sprague-Dawley rats, on restricted food intake, were trained to press a lever for sucrose pellets on a CRF and incrementing discrimination schedule of reinforcement. On the following day, the testing session was followed by an extinction session, where lever pressing was not reinforced. Immediately after extinction, a reversal to CRF was implemented to test for relapse. Trained rats were injected with NT69L (1 mg/kg) or saline 30 min before each testing session. Dopamine, tyrosine 3-hydroxylase, and dopamine receptor mRNA levels were determined. NT69L significantly suppressed the lever pressing behavior for sucrose reinforcement on CRF which measures the "hedonic" value of the reward. NT69L also suppressed sucrose self-administration on the incrementing discrimination schedule of reinforcement (FR3-FR5) that is analogous to the motivational incentive. Reversal to CRF was significantly reduced by pretreatment with NT69L. The suppression of sucrose self administration behavior by pretreatment with NT69L had a pattern similar to that for extinction. The effect of NT69L on dopamine, tyrosine 3-hydroxylase, and dopamine receptor mRNA levels is discussed relative to changes occurring during extinction. PMID- 17113053 TI - Anticonvulsant effects of focal and intracerebroventricular adenosine on penicillin-induced epileptiform activity in rats. AB - Adenosine has potent anticonvulsant effects on various models of experimental epilepsy. In the present study, we examined the effects of focal and intracerebroventricular (i.c.v.) adenosine on penicillin-induced epileptiform activity in Wistar rats. The effects of theophylline, a non-selective adenosine receptor antagonist, were also researched. The recordings of electrocorticogram (ECoG) were carried out by using a data acquisition system, under urethane anesthesia. Adenosine was given in doses of 1, 10 and 100 microg/rat via focal and i.c.v. 30 min after penicillin administration. Theophylline was injected in doses of 1, 10 and 100 microg/rat by i.c.v. too. Adenosine administration significantly decreased the spike frequency while theophylline increased. Focal adenosine is more effective than i.c.v. adenosine. 100 microg adenosine is an effective dose that causes a decrease in epileptiform activity during experiments. We also demonstrated that 100 microg theophylline significantly increased epileptiform activity. Our findings suggest that focal adenosine is more effective than i.c.v. adenosine on epileptiform activity. PMID- 17113054 TI - Nrf2 gene deletion fails to alter psychostimulant-induced behavior or neurotoxicity. AB - The transcription factor NF-E2-related factor (Nrf2) regulates the induction of phase 2 detoxifying enzymes by oxidative stress, including synthesis of the catalytic subunit (xCT) of the heterodimeric cystine-glutamate exchanger (system xc-). Repeated cocaine treatment in rats causes persistent neuroadaptations in glutamate neurotransmission in the nucleus accumbens that result, in part, from reduced activity of system xc-. Since in vitro under- or over-expression of Nrf2 regulates system xc- activity and xCT content, it was hypothesized that in vivo deletion of the Nrf2 gene would: 1) decrease system xc- activity, 2) produce a behavioral phenotype resembling that elicited by chronic cocaine administration, and 3) enhance dopamine depletion after methamphetamine-induced oxidative stress. In all three experiments no genotypic difference was measured between mice sustaining homozygous Nrf2 gene deletion and wild-type littermates. Thus, while Nrf2 is a transcriptional regulator of xCT and capable of protecting cells from oxidative stress, following Nrf2 gene deletion this role can be partially compensated by other mechanisms and methamphetamine-induced oxidative stress and dopamine toxicity does not significantly involve Nrf2. PMID- 17113055 TI - Nerve growth factor eye drop administrated on the ocular surface of rodents affects the nucleus basalis and septum: biochemical and structural evidence. AB - It has been shown that conjunctivally applied NGF in rats can reach the retina and optic nerve. Whether topical eye NGF application reaches the central nervous system is not known. In the present study, we have addressed this question. It was found that topical eye NGF application affects brain cells. Time-course studies revealed that repeated NGF application leads to high concentration of this neurotrophins after 6 h and normal levels after 24 h. Our results also showed that topical eye application of NGF causes an enhanced expression of NGF receptors and ChAT immunoreactivity in forebrain cholinergic neurons, suggesting that ocular NGF application could have a functional role on damaged brain cells. The present findings suggest that eye NGF application can represent an alternative route to prevent degeneration of NGF-receptive neurons involved in disorders such as Alzheimer and Parkinson. PMID- 17113056 TI - Estimated distribution of specific membrane resistance in hippocampal CA1 pyramidal neuron. AB - It has been suggested that dendritic membrane properties play an important role in a synaptic integration. In particular, the specific membrane resistance, one of membrane properties, has been reported to be non-uniformly distributed in a single neuron, although the spatial distribution of the specific membrane resistance is still unclear. To reveal its non-uniformity in dendrite, we estimated the spatial distribution of specific membrane resistance in a single neuron, based on voltage imaging data, observed optically in hippocampal CA1 slices. As the optically recorded data, we used bi-directional propagations of subthreshold excitatory postsynaptic potentials in dendrite, which were not be reproduced numerically with uniform-specific membrane resistance. By numerical simulations for multi-compartment models with non-uniformity of specific membrane resistance, we estimated that the distribution obeys a step function; the optically recorded data were consistently reproduced for the distribution with a steep decrease in the specific membrane resistance at the distal apical dendrite, which occurs 300-500 microm away from the soma. In the estimated distribution, the specific membrane resistance at the distal side is less than about 10(3) Omegacm(2), whereas the resistance at the proximal side is greater than about 10(4) Omegacm(2). This result implies that the specific membrane resistance decreases drastically at the distal apical dendrite in hippocampal CA1 pyramidal neuron. PMID- 17113057 TI - Decreased NR1, NR2A, and SAP102 transcript expression in the hippocampus in bipolar disorder. AB - OBJECTIVES: Schizophrenia is associated with dysfunction of glutamatergic neurotransmission, and several studies have suggested glutamatergic abnormalities in bipolar disorder. Recent data suggest involvement of the NMDA receptor signaling complex, which includes NMDA receptor subunits as well as associated intracellular interacting proteins critical for NMDA receptor assembly, trafficking, and activation; the most well-characterized being PSD93, PSD95, SAP102, and NF-L. Previously, studies from our laboratories have described changes in glutamate receptor subunit transcript and binding site expression in schizophrenia and changes in NMDA receptor binding site expression in bipolar disorder in postmortem brain tissue. In the present work, we focus on the expression of these molecules in hippocampus in schizophrenia and bipolar affective disorder I. METHODS: We performed in situ hybridization to assess hippocampal expression of the transcripts encoding NMDA receptor subunits NR1, 2A, 2B, 2C and 2D, and the transcripts for the NMDA receptor associated PSD proteins PSD95, PSD93, NF-L, and SAP102 in subjects with schizophrenia, bipolar affective disorder I, and a comparison group. We also measured [(3)H]CGP39653 and [(3)H]MK-801 binding site expression in the hippocampus in schizophrenia. RESULTS: There was a significant decrease in the expression of transcripts for NR1 and NR2A subunits and SAP102 in bipolar disorder. We did not detect any changes in these transcripts or in binding site expression in the hippocampus in schizophrenia. CONCLUSIONS: We propose that the NMDA receptor signaling complex, including the intracellular machinery that is coupled to the NMDA receptor subunits, is abnormal in the hippocampus in bipolar disorder. These data suggest that bipolar disorder might be associated with abnormalities of glutamate-linked intracellular signaling and trafficking processes. PMID- 17113058 TI - Antioxidant and radioprotective effect of the active fraction of Pilea microphylla (L.) ethanolic extract. AB - The ethanolic extract of Pilea microphylla (L.) was defatted, successively fractionated with acetone and the residue so obtained was found to be most potent when subjected to detailed free radical scavenging and in vivo radioprotection studies. The most active fraction reacts with free radicals, such as DPPH (50 microM), ABTS(.)(-) (100 microM) and (.)OH (generated by Fenton reaction) with IC(50) value of 23.15 microg/ml, 3.0 microg/ml and 310 microg/ml, respectively. The most active fraction inhibited iron-induced lipid peroxidation in phosphatidyl choline liposomes with an IC(50) of 13.74 microg/ml. The kinetics of scavenging of DPPH and ABTS(.)(-) radicals were followed at different concentrations of the fraction by employing stopped-flow studies. The observed first order decay rate constants at 200 microg/ml and 50 microg/ml of fraction with DPPH (50 microM) and ABTS(.)(-) (50 microM) were found to be 0.4s(-1) and 2.1s(-1), respectively. The fraction when screened for in vivo radioprotection in Swiss albino mice showed 80% protection at a dose of 900 mg/kg and with a DRF of about 1.12. The fraction was also found to protect livers of irradiated mice from depletion of endogenous antioxidant enzymes like glutathione, GST, SOD, catalase and thiols. The fraction also protected the villi height, increased the number of crypt cells while offering general protection to the intestine from acute radiation effects. The fraction also protected the hematopoietic system as assessed by endogenous spleen colony assay, contributing to the overall radioprotective ability. PMID- 17113059 TI - Effect of the 252A>G polymorphism of the lymphotoxin-alpha gene on inflammatory markers of response to cigarette smoking in Korean healthy men. AB - BACKGROUND: The systemic inflammatory response is heightened in smokers. We examined whether the established cardiovascular risk factor, smoking status, might interact with the lymphotoxin-alpha (LTA) gene 252A>G polymorphism in determining concentrations of TNF-alpha and eventually IL-6, adiponectin and CRP downstream in the inflammatory cascade. METHODS: We measured anthropometric parameters, serum lipid profile, glucose, TNF-alpha, IL-6, CRP, adiponectin and urinary excretion of 8-epi PGF2alpha as well as a genotyping for 252A>G polymorphism of LTA in 480 healthy Korean men. RESULTS: After adjustment for age, 208 smokers with an average consumption of 18+/-1 cigarettes/d had higher concentrations of TNF-alpha, IL-6, CRP and urinary excretion of 8-epi PGF2alpha than nonsmokers (n=272). Nonsmokers with G/G had higher TNF-alpha and 8-epi PGF2alpha concentrations than those with A/A or A/G. TNF-alpha concentrations were higher in smokers than nonsmokers of the same genotype. Smokers with G/G showed higher TNF-alpha concentration than those with A/A and had higher IL-6 and urinary 8-epi PGF2alpha concentrations than those with A/G or A/A. Furthermore, smokers carrying the G allele showed lower adiponectin concentrations than those with A/A. There are main effects of genotype and smoking, as well as the smoking genotype interaction to TNF-alpha concentration. CONCLUSION: Our results suggest that the LTA 252A>G polymorphism may modulate the inflammatory effects and oxidative stress of smoking. The detrimental effect of smoking is most clearly seen in men with G/G, suggesting a genotype-specific interaction with smoking. PMID- 17113060 TI - Evaluation of the IRMA TRUpoint and i-STAT creatinine assays. AB - BACKGROUND: The i-STAT (Abbott Diagnostics, East Windsor, NJ) and IRMA TRUpoint (ITC, Edison, NJ) POCT analyzers were evaluated in an oncology center. METHODS: Precision and agreement with our core laboratory creatinine was judged by comparison of 50 consecutive chemotherapy patient results against the Roche rate blanked Jaffe and enzymatic creatinine methods. Glomerular filtration rate (GFR) was estimated using the Cockroft-Gault (CG) calculation and Modification of Diet in Renal Disease Study (MDRD) equation. RESULTS: Precision varied from 1% (enzymatic)-6.1% (TRUpoint). Correlation was good (r>0.9948) with slopes within 5% of the Jaffe and enzymatic methods. Intercepts were <15.9 micromol/l (<0.18 mg/dl), and statistically significant bias (p<0.0025) was noted between the mean of patient specimens for i-STAT correlations to both the Jaffe and enzymatic laboratory creatinine methods. There was statistically significant concordance of estimated GFR between all methods, however, the agreement of estimated GFR to either the Jaffe or enzymatic creatinine laboratory methods was better for the TRUpoint (by either MDRD or CG estimation) and i-STAT (by MDRD equation) (Kappa>0.60) than the i-STAT (by CG estimation) (Kappa=0.41-0.60). CONCLUSION: Small biases in the calibration of analytical creatinine methods can lead to differences in clinical concordance using estimated GFR. Selecting an optimal POCT method depends on the institution's current creatinine method and tolerance for analytical performance and clinical concordance. PMID- 17113062 TI - Isoelectric point determination of cardiac troponin I forms present in plasma from patients with myocardial infarction. AB - BACKGROUND: Cardiac troponin I (cTnI) is a specific marker of myocardial injury. In blood of patients with cardiovascular diseases, cTnI is released as a mixture of free, complexed and post-translationally modified forms. METHODS: The cTnI forms present in the plasma from 8 patients with acute myocardial infarction (AMI) have been analysed by two-dimensional gel electrophoresis (2-DE) and Western Blot using anti-cTnI mAb 19C7 and anti-phosphorylated cTnI (Serines 22 23) mAb 5E6. RESULTS: After immunoextraction of cTnI in plasma samples by 19C7 and 2-DE separation, 4 different forms were detected by 19C7 in 7 out the 8 AMI plasma samples. Two 29 kDa spots corresponding to intact free cTnI forms were detected at pIs 5.2 and 5.4. However, spot with pI 5.4 was also recognized by mAb 5E6, and should be bis-phosphorylated cTnI. Two 55 kDa spots with pIs 6.6 and 6.7 could be IC complexes. CTnI forms with pIs lower than the theoretical pI were also found in free cTnI and phosphorylated cTnI purified materials. CONCLUSIONS: 2-DE analysis of AMI plasma showed the presence of acidic cTnI forms, one of them being phosphorylated. The clinical significance of these forms has to be further investigated. PMID- 17113061 TI - Do mutations causing low HDL-C promote increased carotid intima-media thickness? AB - BACKGROUND: Although observational data support an inverse relationship between high-density lipoprotein (HDL) cholesterol and coronary heart disease (CHD), genetic HDL deficiency states often do not correlate with premature CHD. METHODS: Carotid intima-media thickness (cIMT) measurements were obtained in cases comprising 10 different mutations in LCAT, ABCA1 and APOA1 to further evaluate the relationship between low HDL resulting from genetic variation and early atherosclerosis. RESULTS: In a 1:2 case-control study of sex and age-related (+/ 5 y) subjects (n=114), cIMT was nearly identical between cases (0.66+/-0.17 cm) and controls (0.65+/-0.18 cm) despite significantly lower HDL cholesterol (0.67 vs. 1.58 mmol/l) and apolipoprotein A-I levels (96.7 vs. 151.4 mg/dl) (P<0.05) CONCLUSIONS: Genetic variants identified in the present study may be insufficient to promote early carotid atherosclerosis. PMID- 17113063 TI - Lexical restructuring in the absence of literacy. AB - Vocabulary growth was suggested to prompt the implementation of increasingly finer-grained lexical representations of spoken words in children (e.g., [Metsala, J. L., & Walley, A. C. (1998). Spoken vocabulary growth and the segmental restructuring of lexical representations: precursors to phonemic awareness and early reading ability. In J. L. Metsala & L. C. Ehri (Eds.), Word recognition in beginning literacy (pp. 89-120). Hillsdale, NJ: Erlbaum.]). Although literacy was not explicitly mentioned in this lexical restructuring hypothesis, the process of learning to read and spell might also have a significant impact on the specification of lexical representations (e.g., [Carroll, J. M., & Snowling, M. J. (2001). The effects of global similarity between stimuli on children's judgments of rime and alliteration. Applied Psycholinguistics, 22, 327-342.]; [Goswami, U. (2000). Phonological representations, reading development and dyslexia: Towards a cross-linguistic theoretical framework. Dyslexia, 6, 133-151.]). This is what we checked in the present study. We manipulated word frequency and neighborhood density in a gating task (Experiment 1) and a word-identification-in-noise task (Experiment 2) presented to Portuguese literate and illiterate adults. Ex-illiterates were also tested in Experiment 2 in order to disentangle the effects of vocabulary size and literacy. There was an interaction between word frequency and neighborhood density, which was similar in the three groups. These did not differ even for the words that are supposed to undergo lexical restructuring the latest (low frequency words from sparse neighborhoods). Thus, segmental lexical representations seem to develop independently of literacy. While segmental restructuring is not affected by literacy, it constrains the development of phoneme awareness as shown by the fact that, in Experiment 3, neighborhood density modulated the phoneme deletion performance of both illiterates and ex illiterates. PMID- 17113064 TI - Wheeze detection based on time-frequency analysis of breath sounds. AB - Abnormal breath sounds like wheezes are observed in patients with obstructive pulmonary diseases. The aim of this study was to construct an automatic technique for wheeze detection and monitoring using spectral analysis. Wheezes from 13 patients with diagnosed asthma, chronic obstructive pulmonary disease and pneumonia were recorded and a time-frequency wheeze detector (TF-WD) based on TF wheeze characteristics was constructed. The TF-WD was evaluated using 337 wheezes by comparing its findings with those from clinical auscultation performed by two experts. In addition, the TF-WD was tested against artificial noise. The experimental and testing results justified the efficient performance and high noise robustness of the TF-WD. PMID- 17113065 TI - Craniosynostosis caused by Axin2 deficiency is mediated through distinct functions of beta-catenin in proliferation and differentiation. AB - Targeted disruption of Axin2 in mice induces skeletal defects, a phenotype resembling craniosynostosis in humans. Premature fusion of cranial sutures, caused by deficiency in intramembranous ossification, occurs at early postnatal stages. Axin2 negatively regulates both expansion of osteoprogenitors and maturation of osteoblasts through its modulation on Wnt/beta-catenin signaling. We investigate the dual role of beta-catenin to gain further insights into the skull morphogenetic circuitry. We show that as a transcriptional co-activator, beta-catenin promotes cell division by stimulating its target cyclin D1 in osteoprogenitors. Upon differentiation of osteoprogenitors, BMP signaling is elevated to accelerate the process in a positive feedback mechanism. This Wnt dependent BMP signal dictates cellular distribution of beta-catenin. As an adhesion molecule, beta-catenin promotes cell-cell interaction mediated by adherens junctions in mature osteoblasts. Finally, haploid deficiency of beta catenin alleviates the Axin2-null skeletal phenotypes. These findings support a model for disparate roles of beta-catenin in osteoblast proliferation and differentiation. PMID- 17113066 TI - The ELT-2 GATA-factor and the global regulation of transcription in the C. elegans intestine. AB - A SAGE library was prepared from hand-dissected intestines from adult Caenorhabditis elegans, allowing the identification of >4000 intestinally expressed genes; this gene inventory provides fundamental information for understanding intestine function, structure and development. Intestinally expressed genes fall into two broad classes: widely-expressed "housekeeping" genes and genes that are either intestine-specific or significantly intestine enriched. Within this latter class of genes, we identified a subset of highly expressed highly-validated genes that are expressed either exclusively or primarily in the intestine. Over half of the encoded proteins are candidates for secretion into the intestinal lumen to hydrolyze the bacterial food (e.g. lysozymes, amoebapores, lipases and especially proteases). The promoters of this subset of intestine-specific/intestine-enriched genes were analyzed computationally, using both a word-counting method (RSAT oligo-analysis) and a method based on Gibbs sampling (MotifSampler). Both methods returned the same over-represented site, namely an extended GATA-related sequence of the general form AHTGATAARR, which agrees with experimentally determined cis-acting control sequences found in intestine genes over the past 20 years. All promoters in the subset contain such a site, compared to <5% for control promoters; moreover, our analysis suggests that the majority (perhaps all) of genes expressed exclusively or primarily in the worm intestine are likely to contain such a site in their promoters. There are three zinc-finger GATA-type factors that are candidates to bind this extended GATA site in the differentiating C. elegans intestine: ELT-2, ELT-4 and ELT-7. All evidence points to ELT-2 being the most important of the three. We show that worms in which both the elt-4 and the elt-7 genes have been deleted from the genome are essentially wildtype, demonstrating that ELT-2 provides all essential GATA-factor functions in the intestine. The SAGE analysis also identifies more than a hundred other transcription factors in the adult intestine but few show an RNAi-induced loss-of-function phenotype and none (other than ELT-2) show a phenotype primarily in the intestine. We thus propose a simple model in which the ELT-2 GATA factor directly participates in the transcription of all intestine-specific/intestine-enriched genes, from the early embryo through to the dying adult. Other intestinal transcription factors would thus modulate the action of ELT-2, depending on the worm's nutritional and physiological needs. PMID- 17113067 TI - Adrenergic receptors in the nucleus accumbens shell differentially modulate dopamine and acetylcholine receptor-mediated turning behaviour. AB - The role of alpha- and beta-adrenoceptors in the nucleus accumbens shell in turning behaviour of rats was investigated. Unilateral injections of the alpha adrenoceptor agonist (phenylephrine; 10 microg) and antagonist (phentolamine; 10 microg) as well as the beta-adrenoceptor agonist (isoprenaline; 1 microg) and antagonist (propranolol; 5 microg) into the nucleus accumbens shell did not produce turning behaviour more than that of control vehicle injection. Unilateral injection of a mixture of dopamine D(1) ((+/-)-1-phenyl-2,3,4,5-tetrahydro-1H-3 benzazepine-7,8-diol, SKF 38393; 5 microg) and D(2) (quinpirole; 10 microg) receptor agonists into the nucleus accumbens shell has been found to elicit contraversive pivoting. Such pivoting was dose-dependently inhibited by phenylephrine (5, 10 microg), injected into the nucleus accumbens shell, and the inhibitory effect of phenylephrine (10 microg) was antagonised by phentolamine (10 microg) that per se had no effect on this pivoting. Isoprenaline (0.5, 1 microg) dose-dependently increased the contraversive pivoting induced by the mixture of SKF 38393 (1 microg) and quinpirole (10 microg) injected into the nucleus accumbens shell. The effect of isoprenaline (1 microg) was antagonised by propranolol (5 microg) that per se had no effect on this pivoting. It is concluded that stimulation of accumbal alpha-adrenoceptors inhibits the dopamine dependent pivoting in contrast to stimulation of accumbal beta-adrenoceptors that facilitates this dopamine-dependent pivoting. Unilateral injection of the acetylcholine receptor agonist carbachol (5 microg) into the nucleus accumbens shell has been found to elicit contraversive circling. Such circling was significantly reduced by accumbal administration of either phenylephrine (10, 20 microg) or phentolamine (5, 10 microg) in a dose-independent manner; moreover, both drugs potentiated, but did not counteract, each other's effects. Carbachol induced circling was also reduced by propranolol (2.5, 5 microg), but again in an aspecific manner. It is concluded that alpha- and beta-adrenergic agents have an effect on accumbal acetylcholine receptor-mediated circling through a non adrenergic mechanism. The impact of the present study for putative new treatments of various neuropsychiatric and neurological disorders is discussed. PMID- 17113068 TI - Correlations between acetylcholinesterase inhibition, acetylcholine levels and EEG changes during perfusion with neostigmine and N6-cyclopentyladenosine in rat brain. AB - Organophosphate poisoning can result in seizures and subsequent neuropathology. In order to improve treatment strategies in organophosphate intoxication, the relationship between acetylcholinesterase inhibition, extracellular levels of acetylcholine, and electroencephalogram (EEG) changes was investigated during local perfusion of the reversible acetylcholinesterase inhibitor neostigmine in the hippocampus and striatum of freely moving rats. Acetylcholinesterase activity and acetylcholine levels were measured by microdialysis, and EEG signals were recorded from an electrode placed near the microdialysis probe. A non-linear relationship between the acetylcholinesterase activity and the extracellular amount of acetylcholine was found, the latter being approximately three times higher in the striatum than in the hippocampus upon infusion with 10(-4) M neostigmine. Highly accumulated extracellular acetylcholine significantly correlated with significant relative power increases of the EEG-gamma2-band and a significant relative power decrease in the beta2-band. Co-infusion of the adenosine A1 agonist N6-cyclopentyladenosine partly prevented acetylcholine accumulation, rendered both powers towards control values, and abolished the acetylcholine-EEG correlation. In view of the latter relationship, it is concluded that prevention of acetylcholine accumulation as a concept for neuroprotection in case of organophosphate poisoning, is worth to be further investigated. PMID- 17113069 TI - Use of scopoletin to inhibit the production of inflammatory cytokines through inhibition of the IkappaB/NF-kappaB signal cascade in the human mast cell line HMC-1. AB - Scopoletin (6-methoxy-7-hydroxycoumarin) is a coumarin compound and a pharmacologically active agent that has been isolated from several plant species. However, as yet there is no clear explanation of how scopoletin affects the production of inflammatory cytokine. We therefore used cells from the human mast cell line (HMC-1) to investigate this effect. Scopoletin significantly and dose dependently inhibits the way in which phorbol 12-myristate 13-acetate (PMA) plus A23187 induces the production of inflammatory cytokines such as tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-8 (P<0.05). The maximal rates at which scopoletin (0.2 mM) inhibited the production of TNF-alpha, IL-6, and IL-8 were 41.6%+/-4.2%, 71.9%+/-2.5%, and 43.0%+/-5.7%, respectively. In activated HMC 1 cells, the expression level of nuclear factor (NF)-kappaB/Rel A protein was increased in the nucleus whereas the level of NF-kappaB/Rel A in nucleus was decreased by treatment with scopoletin. Scopoletin decreased PMA plus A23187 induced luciferase activity. Scopoletin also inhibits IkappaBalpha phosphorylation and degradation in cytoplasm. These results indicate that scopoletin has a potential regulatory effect on inflammatory reactions that are mediated by mast cells. PMID- 17113070 TI - Effects of nicotine and vitamin E on glutathione reductase activity in some rat tissues in vivo and in vitro. AB - Effects of nicotine, and nicotine+vitamin E on glutathione reductase (Glutathione: NADP(+) oxidoreductase, EC 1.8.1.7) activity in the muscle, heart, lungs, testicles, kidney, stomach, brain and liver tissues were investigated in vivo and also in vitro. The groups were: nicotine [0.5 mg/kg/day, intraperitoneal (i.p.)]; nicotine+vitamin E [75 mg/kg/day, intragastric (i.g.)]; and control group (receiving only vehicles). There were eight rats per group and supplementation period was 3 weeks. The results showed that nicotine (0.5 mg/kg, i.p.) inhibited glutathione reductase activity significantly in the liver, lungs, heart, stomach, kidney, and testicles by approximately 61.5%, approximately 65%, approximately 70.5%, approximately 72.5%, approximately 64% and approximately 71.5%, respectively, while it had activated glutathione reductase activity in the brain by approximately 11.8%, and had no effect on the muscle glutathione reductase activity. Vitamin E supplementation prevented this nicotine-induced decrease in glutathione reductase activity in liver, lungs, heart, stomach, and kidney. However, it did not prevent this nicotine-induced decrease in testicles. In vitro studies were also carried out to elucidate the effects of nicotine and vitamin E on glutathione reductase activity. In vitro results correlated well with in vivo experimental results in liver, lungs, heart, stomach, and testicular tissues. These results show that vitamin E administration generally restores the inactivation of glutathione reductase activity due to nicotine administration in various rat tissues in vivo, and also in vitro. PMID- 17113071 TI - Endothelium-independent relaxation to raloxifene in porcine coronary artery. AB - Although the vascular action of raloxifene has been studied in several vascular beds, the underlying mechanisms are still incompletely understood. The role of endothelium in raloxifene-induced vascular responses was controversial. The present study was designed to examine endothelium-independent effects of raloxifene in isolated porcine left circumflex coronary arteries. Arterial rings were suspended in organ baths and changes in isometric tension were measured. The large-conductance Ca2+-activated K+(BK(Ca)) currents were recorded using a whole cell patch-clamp technique. Treatment with raloxifene (1-10 micromol/l) reduced the contractions to 9,11-dideoxy-11alpha,9alpha-epoxy-methanoprostaglandin F2alpha (U46619), serotonin (5-HT), endothelin-1 in normal Krebs solution and to CaCl2 in a Ca2+-free, high K+-containing solution. In endothelin-1-contracted rings, raloxifene (0.3 to 50 micromol/l) caused relaxations which were comparable in rings with and without endothelium. The raloxifene-induced relaxation was reduced by putative K+ channel blockers, iberiotoxin and tetraethyl ammonium chloride (TEA+) in rings with and without endothelium, or by elevated extracellular K+ ions (30 mmol/l K+ and 60 mmol/l K+). 13-methyl-7-[9-(4,4,5,5,5 pentafluoropentylsulfinyl)nonyl]-7,8,9,11,12,13,14,15,16, 17-decahydro-6H cyclopenta[a] phenanthrene-3,17-diol (ICI 182,780) did not affect raloxifene induced relaxation. Raloxifene enhanced the outward BK(Ca) currents, which were sensitive to inhibition by iberiotoxin. In summary, the present study shows that raloxifene acutely relaxes porcine coronary arteries via an endothelium independent mechanism without involving the ICI 182,780-sensitive estrogen receptors. Raloxifene mainly acts on the vascular smooth muscle cells to induce vasorelaxation by the inhibition of Ca2+ channels and the activation of BK(Ca) channels. The former mechanism appears to play a more significant role. PMID- 17113072 TI - The inhibitory effect of opioids on HepG2 cells is mediated via interaction with somatostatin receptors. AB - Opioids, acting via G-protein coupled membrane receptors, induce analgesia. However their role is not limited to their anti-nociceptive action. They are found in several peripheral tissues acting as negative regulators of cellular processes. Even though that is not fully elucidated, it becomes obvious that opioids exert their effects in close relation to other neuropeptides such as somatostatin. Hepatocellular carcinoma is one tumor, among others, which secrete bioactive peptides while somatostatin analogs exert an inhibitory effect. We have used the human hepatocyte-derived cancer cell line HepG2, in order to examine the effect of opioids on cell growth and their possible mode of action. Our results show that the opioid ethylketocyclazocine (EKC) inhibits cell proliferation and induces apoptosis. This inhibitory effect is not exerted via opioids receptors since it was not reversed by the opioid antagonist diprenorphine and functional opioid receptors were not found on HepG2 cells. On the contrary, we show that EKC binds to somatostatin receptors, and activates a PTP signalling cascade. In this respect, the interaction of opioids with somatostatin receptors on hepatocellular carcinoma cells, and the fact that they are widely used for pain control, may provide some additional clues for the discrepancies during treatment with somatostatin analogues. PMID- 17113073 TI - Functional roles of muscarinic M2 and M3 receptors in mouse stomach motility: studies with muscarinic receptor knockout mice. AB - Functional roles of muscarinic acetylcholine receptors in the regulation of mouse stomach motility were examined using mice genetically lacking muscarinic M(2) receptor and/or M(3) receptor and their corresponding wild-type (WT) mice. Single application of carbachol (1 nM-30 microM) produced concentration-dependent contraction in antral and fundus strips from muscarinic M(2) receptor knockout (M(2)R-KO) and M(3) receptor knockout (M(3)R-KO) mice but not in those from M(2) and M(3) receptors double knockout (M(2)/M(3)R-KO) mice. A comparison of the concentration-response curves with those for WT mice showed a significant decrease in the negative logarithm of EC(50) (pEC(50)) value (M(2)R-KO) or amplitude of maximum contraction (M(3)R-KO) in the muscarinic receptor-deficient mice. The tonic phase of carbachol-induced contraction was decreased in gastric strips from M(3)R-KO mice. Antagonistic affinity for 4-diphenylacetoxy-N-methyl piperidine (4-DAMP) or 11-([2-[(diethylamino)methyl]-1-piperdinyl]acetyl)-5,11 dihydro-6H-pyrido[2,3-b][1,4]benzodiazepine-6-one (AF-DX116) indicated that the contractile responses in M(2)R-KO and M(3)R-KO mice were mediated by muscarinic M(3) and M(2) receptors, respectively. Electrical field stimulation (EFS, 0.5-32 Hz) elicited frequency-dependent contraction in physostigmine- and N(omega)-nitro L-arginine methylester (l-NAME)-treated fundic and antral strips from M(2)R-KO and M(3)R-KO mice, but the cholinergic contractile components decreased significantly compared with those in WT mice. In gastric strips from M(2)/M(3)R KO mice, cholinergic contractions elicited by EFS were not observed but atropine resistant contractions were more conspicuous than those in gastric strips from WT mice. Gastric emptying in WT mice and that in M(2)/M(3)R-KO mice were comparable, suggesting that motor function of the stomach in the KO mice did not differ from that in the WT mice. The results indicate that both muscarinic M(2) and M(3) receptors but not other subtypes mediate carbachol- or EFS-induced contraction in the mouse stomach but that the contribution of each receptor to concentration response relationships is distinguishable. Although there was impairment of nerve mediated cholinergic responses in the stomach of KO mice, gastric emptying in KO mice was the same as that in WT mice probably due to the compensatory enhancement of the non-cholinergic contraction pathway. PMID- 17113074 TI - Trishomocubanes: novel sigma ligands modulate cocaine-induced behavioural effects. AB - Trishomocubane analogues TC1 (N-(3'-fluorophenyl)ethyl-4-azahexacyclo [5.4.1.0(2,6).0(3,10).0(5,9).0(8,11)]dodecan-3-ol) and TC4 (N-(3' fluorophenyl)methyl-4-azahexacyclo [5.4.1.0(2,6).0(3,10).0(5,9).0(8,11)]dodecan-3 ol) were evaluated for their modulatory effects on locomotor activity as well as interactions with cocaine-induced responses. TC1 and TC4 have high affinity and moderate to high selectivity for sigma(1) (Ki=10 nM, sigma1/sigma2=0.03) and sigma2 (Ki=20 nM, sigma1/sigma2=7.6) receptor subtypes respectively. Both compounds have negligible affinity for the dopamine (DAT), serotonin (SERT), and norepinephrine (NET) transporters. In behavioural studies, TC1 produced a dose related inhibition in spontaneous locomotor activity measured in a Digiscan apparatus. TC1 attenuated the stimulatory locomotor effect of 20 mg/kg cocaine with a half-maximal depressant activity (ID50) of 38.6 mg/kg. TC1 (dose range of 25 to 100 mg/kg) also partially substituted for the effect of cocaine (10 mg/kg) in a discriminative stimulus task, involving the trained discrimination between cocaine and saline using a two-lever choice method. Following a dose of 50 mg/kg TC1, a maximum of 31% substitution was reached. The response rate was reduced to 56% of vehicle control following a TC1 dose of 100 mg/kg. These behavioural effects suggest that TC1 can act as an antagonist via the sigma1 receptor. In contrast to TC1, TC4 produced a stimulant effect in locomotor activity with the ED50 estimated at 0.94 mg/kg. In addition, TC4 failed to inhibit cocaine-induced stimulation; neither did it substitute for the discriminative stimulus effects of cocaine. TC4 thus appears to interact predominantly with the sigma2 receptor subtype (sigma1/sigma2=7.6) which may result in dopamine stimulation independent of the effects of cocaine. The differential effect of TC1 and TC4 warrants further study of the mechanism of these actions. Present data also suggests a potential role for trishomocubane analogues in developing medication or research tools for cocaine addiction. PMID- 17113075 TI - Interactive effects of the mGlu5 receptor antagonist MPEP and the mGlu2/3 receptor antagonist LY341495 on nicotine self-administration and reward deficits associated with nicotine withdrawal in rats. AB - Stimulatory actions of nicotine on mesocorticolimbic dopamine transmission are partly mediated by nicotine-induced glutamate release acting on ionotropic and metabotropic glutamate (mGlu) receptors. Because both presynaptic inhibitory mGlu2/3 and postsynaptic excitatory mGlu5 receptors provide potential targets for treatment of aspects of nicotine dependence, we examined interacting effects of mGlu5 (2-methyl-6-(phenylethynyl)-pyridine, MPEP) and mGlu2/3 (LY341495) receptor antagonists on nicotine self-administration and brain reward threshold elevations associated with spontaneous nicotine withdrawal in rats. We hypothesized that increasing glutamate transmission by blocking presynaptic inhibitory mGlu2/3 autoreceptors would antagonize MPEP-induced decreases in nicotine self administration. We also hypothesized that blocking postsynaptic actions of glutamate on mGlu5 receptors would exacerbate nicotine withdrawal-induced reward deficits, and that this effect would be attenuated by co-administration of the mGlu2/3 receptor antagonist LY341495. MPEP selectively decreased nicotine, but not food, self-administration in rats. LY341495 slightly decreased both nicotine and food self-administration. Co-administration of LY341495 with MPEP attenuated the effectiveness of MPEP in decreasing nicotine intake, although MPEP was still effective. Spontaneous nicotine withdrawal induced somatic signs of withdrawal and reward threshold elevations indicating reward deficits. MPEP increased somatic signs and reward deficits in both nicotine- and saline-withdrawing rats. Thus, while mGlu5 receptor antagonists may be therapeutically useful in decreasing tobacco smoking, they worsen nicotine withdrawal. Co-administration of LY341495 reduced MPEP-induced reward deficits in both nicotine- and saline withdrawing rats. Thus, increasing glutamate transmission via mGlu2/3 autoreceptor blockade reduces the effects of mGlu5 receptor blockade on nicotine self-administration and MPEP-induced exacerbation of brain reward deficits associated with nicotine withdrawal. PMID- 17113076 TI - Immunomodulation by interleukin-4 suppresses matrix metalloproteinases and improves cardiac function in murine myocarditis. AB - Immune response is critically involved in determining the course of viral myocarditis and immunomodulation. Different cytokines may have either deleterious or protective effects. Following acute Coxsackievirus B3 infection, intramyocardial inflammation is associated with altered myocardial matrix metalloproteinase (MMP) expression and left ventricular dysfunction. In this study, we evaluated the effect of exogenous interleukin-4 treatment on myocardial inflammation, MMPs and left ventricular function in Coxsackievirus B3-induced acute murine myocarditis. Eight-week-old inbred male BALB/c (H-2d) mice (The Jackson Laboratory, Bar Harbor, Maine, USA) were used. Myocardial inflammation was measured by immunohistochemical detection of CD3(+)-, CD8a(+)-T-lymphocytes, and CD11b+ macrophages. In situ hybridization was used to detect enteroviral genome in the myocardium. Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was employed to detect cytokine and MMP mRNA. MMP activity was quantified by zymography analysis. Detection of myocytolysis was performed by Luxol fast blue staining. In the early acute phase, in comparison to infected mice without treatment, interleukin-4 administration (200 ng daily) reduced intramyocardial inflammation (CD3+ lymphocytes: 55.3+/-7.0 vs. 72.1+/ 13.7 cells/mm2, P < 0.05; CD8a+ lymphocytes: 31.7+/-3.6 vs. 64.2+/-7.7 cells/mm2, P < 0.05; CD11b+ macrophages: 5.1+/-2.3 vs. 13.2+/-2.5 cells/mm2, P < 0.05). It also down-regulated interleukin-2 (IL) (1.7-fold, P < 0.001) but increased transforming growth factor-beta1 (TGF) (1.5-fold, P < 0.001) and IL-4 (1.4-fold, P < 0.001). IL-4 suppressed MMP-2/-3/-9 transcription and activity. These biochemical alterations were accompanied by a significant improvement of left ventricular function as assessed by Milar tip catheter (left ventricular endsystolic pressure, 1.3-fold, P < 0.01; dP/dt max, 1.5-fold, P < 0.01). Immunomodulation by exogenous IL-4 treatment may lead to an anti-inflammatory effect with the inhibition of Th1 cell phenotypic response, which may further mediate the down-regulation of MMPs. A significant suppression of MMPs may mainly contribute to an improvement of left ventricular dysfunction in acute murine CVB3 induced myocarditis. PMID- 17113077 TI - Neuroprotective effects of brimonidine treatment in a rodent model of ischemic optic neuropathy. AB - Ischemic optic neuropathy (ION) is a common disorder caused by disruption of the arterial blood supply to the optic nerve. It can result in significant loss of visual acuity and/or visual field. An ischemic optic nerve injury was produced in rats by intravenous injection of Rose Bengal dye followed by argon green laser application to the retinal arteries overlying the optic nerve, causing a coagulopathy within the blood vessels and disruption of optic nerve and retinal perfusion. The effect of brimonidine tartrate eye drops on survival of retinal ganglion cell axons in this experimental paradigm was studied. One eye was treated and the contralateral eye served as a control. Four groups of animals were used for this study. Group 1 received 7 days of treatment with 0.15% brimonidine tartrate eye drops twice a day prior to the ischemic injury. Group 2 animals received 0.15% brimonidine tartrate eye drops twice a day for 14 days after photocoagulation injury. Animal groups 3 and 4 received eye drops of 0.9% NaCl twice a day either daily for 7 days before injury or daily for 14 days, respectively. All rats were sacrificed 5 months after the injury to ascertain long-term optic axon survival. Coagulopathy-induced optic nerve ischemia resulted in a 71% loss of optic axons. Treatment with brimonidine daily for the 7 days prior to the injury resulted in a greater survival of optic axons, with only a 56.1% loss compared to control. Brimonidine treatment every day for 14 days after the ischemic injury did not result in a significant rescue of optic axons compared to injury alone. In summary, the application of brimonidine eye drops for one week prior to an ischemic injury resulted in a statistically significant increase in survival of optic axons within the injured optic nerves. Brimonidine treatment of the eye after the ischemic injury did not result in axon rescue, and axon loss was similar to the injured optic nerves treated with saline only. These results suggest that brimonidine may have potential use for prevention of ION in at-risk patients. PMID- 17113078 TI - Decreased expression of l-dopa-induced dyskinesia by switching to ropinirole in MPTP-treated common marmosets. AB - Current concepts suggest that pulsatile stimulation of dopamine receptors following L-dopa administration leads to priming for dyskinesia in 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine hydrochloride (MPTP)-treated primates, while continuous dopaminergic stimulation with long-acting dopamine agonists does not. We investigated whether L-dopa-induced dyskinesia is reduced by switching to a dopamine agonist. MPTP-treated marmosets received chronic treatment with L-dopa or ropinirole in doses producing equivalent motor activity and reversal of motor deficits. Administration of L-dopa led to the rapid onset of moderate to severe dyskinesia, whereas ropinirole produced only mild dyskinesia. Animals initially treated with L-dopa were switched to an equivalent dose of ropinirole and those treated with ropinirole were switched to an equivalent dose of L-dopa for 56 days. L-dopa-primed animals that were switched to ropinirole showed a trend towards a reduction of dyskinesia intensity, whereas animals initially treated with ropinirole and switched to L-dopa showed a trend toward increased dyskinesia intensity. A subsequent, acute L-dopa challenge reversed motor deficits and induced intense dyskinesia in both groups. This suggests that L-dopa leads to the priming and expression of dyskinesia, but that expression is not maintained when switching to a long-acting dopamine agonist. In contrast, dopamine agonists may prime for dyskinesia, but do not lead to its full expression. PMID- 17113079 TI - Effects of transcranial direct current stimulation coupled with repetitive electrical stimulation on cortical spreading depression. AB - We have recently shown that two techniques of brain stimulation - repetitive electrical stimulation (ES) (that mimics transcranial magnetic stimulation) and transcranial direct current stimulation (tDCS) - modify the velocity of cortical spreading depression (CSD) significantly. Herein we aimed to study the effects of these two techniques combined on CSD. Thirty-two Wistar rats were divided into four groups according to the treatment: sham tDCS/sham ES, sham tDCS/1 Hz ES, anodal tDCS/1 Hz ES, cathodal tDCS/1 Hz ES. Our findings show that 1 Hz ES reduced CSD velocity, and this effect was modified by either anodal or cathodal tDCS. Anodal tDCS induced larger effects than cathodal tDCS. Hereby CSD velocity was actually increased significantly after anodal tDCS/1 Hz ES. Our results show that combining two techniques of brain stimulation can modify significantly the effects of ES alone on cortical excitability as measured by the neurophysiological parameter of cortical spreading depression and therefore provide important insights into the effects of this new approach of brain stimulation on cortical activity. PMID- 17113080 TI - Entamoeba histolytica: an ecto-phosphatase activity regulated by oxidation reduction reactions. AB - In this work, an ecto-phosphatase activity of Entamoeba histolytica was characterized using intact cells. This activity presented the following biochemical characteristics: (i) it hydrolyzes p-NPP with V(max) of 8.00+/-0.22 nmol p-NP x h(-1) x 10(-5) cells and K(m) of 2.68+/-0.25 mM; (ii) it is inhibited by acid phosphatase inhibitors, such as sodium molybdate (K(i)=1.70+/-0.24 microM) and sodium fluoride (K(i)=0.25+/-0.02 mM); (iii) it also showed high sensitivity to phosphotyrosine phosphatase inhibitors, such as sodium orthovanadate (K(i)=1.07+/-0.14 microM), bpV-PHEN (K(i)=0.38+/-0.02 microM) and mpV-PIC (K(i)=0.39+/-0.04 microM). Zn(2+), an oxidizing agent, decreased the enzymatic activity in 50%. DTT and GSH, two reducing agents, enhanced the activity twofold. The non-invasive E. histolytica and free-living E. moshkovskii were less efficient in hydrolyzing p-NPP than the pathogenic E. histolytica suggesting that this enzyme could represent a virulence marker for this cell. PMID- 17113081 TI - KLF6 degradation after apoptotic DNA damage. AB - Kruppel-like factor 6 (KLF6) is a cancer gene (). Here, we demonstrate that KLF6 protein is rapidly degraded when apoptosis is induced via the intrinsic pathway by cisplatin, adriamycin, or UVB irradiation in multiple cell lines (HCT116, SW40, HepG2, PC3-M, Skov3, NIH-3T3, 293T, GM09706, and MEF, IMR-90). KLF6 degradation occurred in the presence or absence of p53, was associated with ubiquitination, mediated by the proteasome (half-life 16min, unstimulated), and independent of caspases and calpain. KLF6 was unchanged by apoptosis via the extrinsic/death-receptor pathway. Deregulation of KLF6 stability may alter its tumor suppressor function and/or the response of tumors to chemotherapeutics. PMID- 17113082 TI - The mechanism of graft transmission of sense and antisense gene silencing in tomato plants. AB - We investigated the effect of target mRNA level on grafting-transmitted gene silencing in tomato plants by using a strong ACC oxidase 1 (ACO1) silencer as the stock and transgenic ACO1 overexpressers as scions. Manifestation of graft transmission of sense gene silencing required a high initial level of target mRNA in the scion. A relatively high level of siRNA, similar to that in the strong ACO1 silencer, was also detected in the silencing-susceptible strong ACO1 overexpressers prior to grafting. After grafting the silencing signal from the stock enhanced the level of the siRNAs in the scion and the ACO1 mRNA level was reduced dramatically. Using stock and scions producing different siRNAs we provided evidence that the transmissible silencing signal does not correspond to the bulk siRNAs in the stock. We also showed, contrary to a previous report, that antisense silencing was graft-transmissible but it took longer to manifest itself. The delay in graft transmission from antisense-silenced plants could be attributed to the difference in the nature or strength of the signal or the mechanism of its amplification, but is further evidence of mechanistic similarities between sense and antisense silencing. PMID- 17113083 TI - Altered beta-secretase enzyme kinetics and levels of both BACE1 and BACE2 in the Alzheimer's disease brain. AB - beta-Secretase is the rate limiting enzymatic activity in the production of amyloid-beta peptide, the primary component of senile plaque pathology in Alzheimer's disease (AD). This study performed the first comparative analysis of beta-secretase enzyme kinetics in AD and control brain tissue. Results found V(max) values for beta-secretase to be significantly increased, and K(m) values unchanged in AD temporal cortex compared to matched control temporal cortex. The increased V(max) in AD cases, did not correlate with levels of BACE1, and decreased BACE1 and BACE2 levels correlated with the severity of neurofibrillary pathology (I-VI), and synaptic loss in AD. These results indicate that increased V(max) for beta-secretase is a feature of AD pathogenesis and this increase does not correlate directly with levels of BACE1, the principal beta-secretase in brain. PMID- 17113084 TI - Glycosylation regulates turnover of cyclooxygenase-2. AB - Cyclooxygenase-2 (COX-2) catalyzes the rate-limiting step in the prostanoid biosynthesis pathway, converting arachidonic acid into prostaglandin H(2). COX-2 exists as 72 and 74kDa glycoforms, the latter resulting from an additional oligosaccharide chain at residue Asn(580). In this study, Asn(580) was mutated to determine the biological significance of this variable glycosylation. COS-1 cells transfected with the mutant gene were unable to express the 74kDa glycoform and were found to accumulate more COX-2 protein and have five times greater COX-2 activity than cells expressing both glycoforms. Thus, COX-2 turnover appears to depend upon glycosylation of the 72kDa glycoform. PMID- 17113085 TI - Arginase-flotillin interaction brings arginase to red blood cell membrane. AB - Flotillin-1 and arginase are both up-regulated in red blood cell membrane of type 2 diabetic patients. For studying why the soluble arginase can bind to the membrane and whether such binding would modify arginase activity, the arginase1 and related proteins were cloned and expressed. The results showed that flotillin 1 can interact with arginase1, and hence arginase activity was up-regulated by 26.8%. It was estimated that about 61% of arginase1 is bound to the membrane mediated by flotillin-1. The arginase activity in diabetic patients was significantly higher than that of the controls (752.4+/-38.5 U/mg protein vs 486.7+/-28.7 U/mg protein). PMID- 17113086 TI - Impact of infertility drugs after treatment of borderline ovarian tumors: results of a retrospective multicenter study. AB - OBJECTIVE: To evaluate safety and fertility outcome after the use of infertility drugs in patients who were treated conservatively for a borderline ovarian tumor (BOT). DESIGN: A retrospective multicenter study. SETTING: Centers participating in the French National Register on In Vitro Fertilization registry. PATIENT(S): Thirty patients who were treated for BOT who underwent ovarian induction (OI). INTERVENTION(S): Ovarian induction was performed in 25 patients for infertility after conservative surgery and before surgery for recurrent disease in 5 patients with a single ovary (emergency cases). MAIN OUTCOMES MEASURE(S): Fertility and recurrences rates. RESULT(S): The mean number of cycles of OI per patient was 2.6 (range, 1-10 cycles). The median follow-up time after treatment of the BOT was 93 months (range, 26-276 months). After a median follow-up time of 42 months after OI, 4 recurrences were observed (initial management was simple cystectomy in 3 of them). All recurrences were borderline tumors on a remaining ovary that had been treated by surgery alone. All patients are currently disease-free. Thirteen pregnancies were observed (10 pregnancies (40%) in the group of 25 patients who were treated for infertility). CONCLUSION(S): These results suggest that infertility drugs could be used safely in patients who experience infertility after conservative management of an early-stage BOT. PMID- 17113087 TI - Immunogenicity of four complementary deoxyribonucleic acid fragments from rabbit zona pellucida 3 and their effects on fertility. AB - OBJECTIVE: To evaluate the exact region in different exons of rabbit zona pellucida (ZP)3 involved in recognition and binding between sperm and the ZP. DESIGN: Prospective study of a female immunocontraceptive. SETTING: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences. ANIMAL(S): BALB/C mice. INTERVENTION(S): Immunization recombinant vaccines. MAIN OUTCOME MEASURE(S): Oocyte immunofluorescence and immunohistochemistry. RESULT(S): The immunogenicity and effects on fertility of these four fragments we used were different. Except for the ZP domain, the other three fragments of rabbit ZP3 may be useful as antigen to elicit antibodies. Antiserum was specific and obvious. The fertility of mice after immunization decreased slightly compared with the control. CONCLUSION(S): The most effective fragment that is associated with the sperm binding was from sequences contained in exons 5-8 or oligosaccharide linked to this region. Exons 5-8 or oligosaccharide linked to this region may exist outside of the ZP matrix and be safe for use as the antigen. The ZP domain may be not related to the recognition and binding. PMID- 17113088 TI - Role of gonadotropin-releasing hormone antagonists in poor responders. AB - OBJECTIVE: To evaluate the role of GnRH antagonists in poor-responder protocols. DESIGN: Literature review. CONCLUSION(S): The optimum stimulation protocol for poor responders is unknown. Although many IVF programs currently use GnRH antagonists for poor responders, there have been only four prospective, randomized trials comparing GnRH antagonists to alternate protocols. None of these studies had sufficient power to evaluate a difference in pregnancy rates (PRs), and in all four cases, IVF outcomes were comparable. Nevertheless, interest in the use of GnRH antagonists in poor responders has continued. GnRH antagonists may be associated with simpler stimulation protocols, lower gonadotropin requirements, reduced patient costs, and shorter downtimes between consecutive cycles. However, the greatest advantage of GnRH antagonists may lie in the ability to assess ovarian reserves immediately prior to deciding whether or not to initiate gonadotropin stimulation. The ability to respond to cycle-to cycle variation in antral follicle counts may allow the optimization of oocyte yield and reduce cycle cancellation rates. It remains to be seen if this approach (initiating gonadotropins only in cycles where an adequate antral follicle count is present) also translates into higher clinical PRs for poor responders. PMID- 17113089 TI - Immunological localization of syndecan-1 in human endometrium throughout the menstrual cycle. AB - OBJECTIVE: To examine the expression of syndecan-1 in the endometrium during the menstrual cycle. DESIGN: The expression of syndecan-1 was determined by tissue microarray and immunohistochemistry. SETTING: Academic clinical and research laboratories. PATIENT(S): Seventy-one regularly cycling women who underwent endometrial biopsy. INTERVENTION(S): Endometrial samples representing five stages of the menstrual cycle were used for the study. MAIN OUTCOME MEASURE(S): Semiquantitative analysis by evaluating the intensity of immunohistochemical reactivity of syndecan-1 by using a modified HSCORE. RESULT(S): Endometrial syndecan-1 was expressed in the luminal and glandular epithelium as well as in the stroma throughout the menstrual cycle in a nonsynchronized fashion. In the luminal epithelium, the expression of syndecan-1 was statistically significantly higher in the mid-secretory phase when compared with the proliferative phase. In the stroma, however, the expression of syndecan-1 was down-regulated after ovulation and remained at a low level through the secretory phases. Differences between the proliferative and mid secretory, as well as between the ovulatory and the early, mid, and late secretory phases, all were statistically significant. CONCLUSION(S): Syndecan-1 is up-regulated in luminal epithelial cells during the mid-secretory phase and is down-regulated in the stroma during the early to late secretory phases. The differential expression of syndecan-1 coincides with the endometrial remodeling throughout the menstrual cycle. PMID- 17113090 TI - Weight loss by bariatric surgery and subsequent fertility. AB - The purpose of this article is to appraise the literature to ascertain whether the use of bariatric surgery should have a role in the contemporary management of the morbidly obese infertile patient before proceeding with infertility treatment. PMID- 17113091 TI - Altered in vitro immune response to hypoxia-treated normal peritoneal fibroblasts. AB - Hypoxia treatment of normal peritoneal fibroblasts results in alterations that are characteristic for fibroblasts obtained from postoperative peritoneal adhesion tissue. In this study, we have shown that hypoxia treatment enhances normal peritoneal fibroblast elimination by lymphokine-activated killer cells to the level of adhesion tissue fibroblasts, suggesting a pivotal role of hypoxia in the adhesion development. PMID- 17113092 TI - A comparison of heterotopic and intrauterine-only pregnancy outcomes after assisted reproductive technologies in the United States from 1999 to 2002. AB - OBJECTIVE: To compare the risk for adverse outcomes of pregnancies between heterotopic (defined as a simultaneous intrauterine and ectopic pregnancy) and intrauterine-only pregnancies achieved through assisted reproductive technologies (ARTs). DESIGN: Retrospective cohort study. SETTING: ART centers in the United States. PATIENT(S): Patients were studied in terms of cycles reported to the population-based United States ART Registry, which included 207 heterotopic and 132,660 intrauterine-only pregnancies reported from 1999 to 2002. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Outcomes of heterotopic and intrauterine-only pregnancies and deliveries (spontaneous abortion, induced abortion, still birth, and live birth). Perinatal outcomes (preterm, low birth weight [LBW], preterm LBW, and term LBW) for live-birth deliveries were also assessed. RESULT(S): Heterotopic pregnancies were more likely to end in spontaneous (relative risk = 2.05; 95% confidence interval, 1.67-2.51) or induced (relative risk = 10.28, 95% confidence interval, 6.76-15.65) abortions than were intrauterine-only pregnancies. There was no significant difference in perinatal outcomes studied, regardless of adjustment for maternal age, infertility diagnosis, previous live births, and type of ART procedure. CONCLUSION(S): Heterotopic pregnancies were more likely to result in spontaneous or induced abortions than were intrauterine-only pregnancies. There was no difference in perinatal outcomes between heterotopic and intrauterine-only pregnancies progressing to live birth. PMID- 17113093 TI - Advancements in transfection technologies for Plasmodium. AB - Malaria is a global problem that affects millions of people annually. A relatively poor understanding of the malaria parasite biology has hindered vaccine and drug development against this disease. Robust methods for genetic analyses in Plasmodium have been lacking due to the difficulties in its genetic manipulation. Introduction of transfection technologies laid the foundation for genetic dissection of Plasmodium and recent years have seen the development of novel tools for genetic manipulation that will help us delineate the intriguing biology of this parasite. This review focuses on such recent advances in transfection technologies for Plasmodium that have improved our ability to carry out more thorough genetic analyses of the biology of the malaria parasite. PMID- 17113094 TI - Isolation and structural analysis of the cyclic fatty acid monomers formed from eicosapentaenoic and docosahexaenoic acids during fish oil deodorization. AB - Long-chain polyunsaturated fatty acids (LC-PUFAs) present in fish oils are thermolabile molecules. Among the degradation reactions encountered, thermal cyclization occurs during refining or other heat treatments. Numerous studies have been carried out in the past to quantify and determine the structures of cyclic fatty acid monomers (CFAMs) formed from oleic, linoleic and linolenic acids in heated vegetable oils. Recently, much attention have been given to LC PUFAs due to their potential health benefits. However, data on quantification of CFAMs formed from these fatty acids, such as eicosapentaenoic acid (EPA, cis-5, cis-8, cis-11, cis-14, cis-17 20:5) and docosahexaenoic acid (DHA, cis-4, cis-7, cis-10, cis-13, cis-16, cis-19 22:6), the two main LC-PUFAs in fish oils, are scarce. In the present study, structural analyses of CFAMs formed from EPA and DHA during the deodorization of fish oil are presented. Fish oil sample was deodorized at 250 degrees C for 3 h under a pressure of 1.5 mbar in a laboratory deodorizer. The CFAMs formed during heat treatment of fish oil were isolated by a combination of saponification, esterification, urea fractionations and column chromatography. Structural analyses of C20- and C22-CFAMs were achieved by gas chromatography electronic-ionization mass-spectrometry (GC-EI-MS) of their 4,4 dimethyloxazoline (DMOX) derivatives. We identified seven out of 13 possible structures of hydrogenated CFAMs formed from EPA, and nine out of 16 possible structures of CFAM formed from DHA. Major CFAMs from both EPA and DHA were cyclohexyl isomers. All possible cyclohexyl isomers were found but only nine out of 18 of the cyclopentyl isomers were present in concentration sufficient for identification. Chemical mechanisms involved in the formation of polyunsaturated LC-PUFAs have been investigated. The results have shown that general principle involved in the cyclization of LC-PUFAs is same as that for the thermal cyclization of oleic, linoleic and alpha-linolenic acids. PMID- 17113095 TI - Simultaneous analysis of polychlorinated biphenyls and organochlorine pesticides in water by headspace solid-phase microextraction with gas chromatography-tandem mass spectrometry. AB - Headspace solid-phase microextraction combined with gas chromatography-ion trap tandem mass spectrometry (HS-SPME-GC-ITMS-MS) method has been developed and studied for the simultaneous determination of 15 organochlorine pesticides (OCPs) and 20 polychlorinated biphenyls (PCBs) in aqueous samples. To perform the HS SPME polydimethylsiloxane (PDMS) (7, 30 and 100 microm film thickness) and polydimethylsiloxane-divinylbenzene (PDMS-DVB) fibers were initially compared on the basis of their absorption capacities for the selected compounds, and PDMS 100 microm film thickness was selected to accomplish the rests of essays. The influence of various parameters on OCPs and PCBs extraction efficiency by HS-SPME was thoroughly studied using GC-electron capture detector (ECD). Parameters such as collision induced dissociation (CID) resonant excitation amplitude and RF storage level were optimized to increase specificity and sensibility for ITMS-MS analysis. The performance of proposed HS-SPME-GC-ITMS-MS methodology with respect to linearity, reproducibility and limit of detection (LOD) was evaluated by water spiked with target compounds. The linear range of most compounds was found to be between 0.01 and 1 ng mL(-1) and the limits of detection were between 0.4 and 26 pg mL(-1). The reproducibility of the method (n = 6), expressed as relative standard deviation (RSD), was between 5 and 21%. Finally, developed procedure was applied to determine selected OCPs and PCBs in river water samples in concentration below 0.1 ng mL(-1) can be easily carried out with ultra selectivity and precision. PMID- 17113096 TI - Voice onset time for female trained and untrained singers during speech and singing. AB - The purpose of this study was to examine the voice onset times of female trained and untrained singers during spoken and sung tasks. Thirty females were digitally recorded speaking and singing short phrases containing the English stop consonants /p/ and /b/ in the word-initial position. Voice onset time was measured for each phoneme and statistically analyzed. Mixed-ANOVAs revealed significantly longer voice onset time durations during speech for /p/ as compared to sung productions. No significant differences between the trained singers and untrained singers were observed. In addition, no task differences occurred for the /b/ productions. The results indicated that the type of phonatory task influences VOT for voiceless stops in females. LEARNING OUTCOMES: As a result of this activity, the reader will be able to (1) understand articulatory and phonatory differences between spoken and sung productions; (2) understand the articulatory and phonatory timing differences between trained singers and untrained singers during spoken and sung productions. PMID- 17113098 TI - Physiological differences between two sugar-sensitive neurons in the galea and the maxillary palp of the spruce budworm larva Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae). AB - The L1 contact-chemoreceptor sensillum on the maxillary palp of the spruce budworm larva Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae) was examined electrophysiologically for its responses to stimulation by various pyranose and furanose sugars. The results were compared to those from previous work on the sugar-sensitive neuron of the LST sensillum on the galea. We show that the L1 contact-chemoreceptor sensillum contains one sugar-sensitive neuron with furanose but no pyranose sites. It has response characteristics that differ from those of the sugar-sensitive neuron in the LST. Behavioural 2-choice feeding experiments show that, even with both known sugar-sensitive neurons disabled, larvae can still discriminate between disks treated with either distilled water or alpha-D-glucose. We conclude that the epipharyngeal sensilla must thus also contain a sugar-sensitive neuron. PMID- 17113097 TI - Method for generation of in vivo biotinylated recombinant antibodies by yeast mating. AB - We describe here a novel method for generation of yeast-secreted, in vivo biotinylated recombinant antibodies, or biobodies. Biobodies are secreted by diploid yeast resulting from the fusion of two haploid yeast of opposite mating type. One yeast carries a cDNA encoding an antibody recognition sequence fused to an IgA1 hinge and a biotin acceptor site (BCCP) at the C-terminus; the other carries a cDNA encoding an E. coli biotin ligase (BirA) fused to KEX2 golgi localization sequences, so that BirA can catalyze the biotin transfer to the recognition sequence-fused BCCP within the yeast secretory compartment. We illustrate this technology with biobodies against HE4, a biomarker for ovarian carcinoma. Anti-HE4 biobodies were derived from clones or pools of anti-HE4 specific yeast-display scFv, constituting respectively monoclonal (mBb) or polyclonal (pBb) biobodies. Anti-HE4 biobodies were secreted directly biotinylated thus bound to labeled-streptavidin and streptavidin-coated surfaces without Ni-purification. Anti-HE4 biobodies demonstrated specificity and sensitivity by ELISA assays, flow cytometry analysis and Western blots prior to any maturation; dissociation equilibrium constants as measured by surface plasmon resonance sensor were of K(d)=4.8 x 10(-9) M and K(d)=5.1 x 10(-9) M before and after Ni-purification respectively. Thus, yeast mating permits cost-effective generation of biotinylated recombinant antibodies of high affinity. PMID- 17113099 TI - A glassy-winged sharpshooter cell line supports replication of Rhopalosiphum padi virus (Dicistroviridae). AB - Rhopalosiphum padi virus (RhPV) (family Dicistroviridae; genus Cripavirus) is an icosahedral aphid virus with a 10kb positive-sense RNA genome. To study the molecular biology of RhPV, identification of a cell line that supports replication of the virus is essential. We screened nine cell lines derived from species within the Lepidoptera, Diptera and Hemiptera for susceptibility to RhPV following RNA transfection. We observed cytopathic effects (CPE) only in cell lines derived from hemipterans, specifically GWSS-Z10 cells derived from the glassy winged sharp shooter, Homalodisca coagulata and DMII-AM cells derived from the corn leaf hopper, Dalbulus maidis. Translation and appropriate processing of viral gene products, RNA replication and packaging of virus particles in the cytoplasm of GWSS-Z10 cells were examined by Western blot analysis, Northern blot hybridization and electron microscopy. Infectivity of the GWSS-Z10 cell derived virus particles to the bird cherry-oat aphid, R. padi, was confirmed by RT-PCR and Western blot. The GWSS-Z10 cell line provides a valuable tool to investigate replication, structure and assembly of RhPV. PMID- 17113100 TI - Endosymbionts of Ctenocephalides felis felis (Siphonaptera: Pulicidae) obtained from dogs captured in Belo Horizonte, Minas Gerais, Brazil. AB - Specimens of fleas Ctenocephalides felis felis (1052 female symbol/448 male symbol), obtained from 150 dogs in Centro de Controle de Zoonoses de Belo Horizonte, Minas Gerais, Brazil, were dissected and examined for endosymbionts. Three protozoan, Nolleria pulicis, a gregarine (Actinocephalidae) and Leptomonas sp., together with one cestode, Dipylidium caninum were identified. Infections by N. pulicis and Leptomonas sp. occurred mainly in the warm-rainy period. The prevalence and distribution of these endosymbionts in fleas derived from Brazil and South America, and the their variation according to sex and season, are reported for the first time. PMID- 17113101 TI - Tandem dimerization of the human p53 tetramerization domain stabilizes a primary dimer intermediate and dramatically enhances its oligomeric stability. AB - Tetramerization of the human p53 tumor suppressor protein is required for its biological functions. However, cellular levels of p53 indicate that it exists predominantly in a monomeric state. Since the oligomerization of p53 involves the rate-limiting formation of a primary dimer intermediate, we engineered a covalently linked pair of human p53 tetramerization (p53tet) domains to generate a tandem dimer (p53tetTD) that minimizes the energetic requirements for forming the primary dimer. We demonstrate that p53tetTD self-assembles into an oligomeric structure equivalent to the wild-type p53tet tetramer and exhibits dramatically enhanced oligomeric stability. Specifically, the p53tetTD dimer exhibits an unfolding/dissociation equilibrium constant of 26 fM at 37 degrees C, or a million-fold increase in stability relative to the wild-type p53tet tetramer, and resists subunit exchange with monomeric p53tet. In addition, whereas the wild type p53tet tetramer undergoes coupled (i.e. two-state) dissociation/unfolding to unfolded monomers, the p53tetTD dimer denatures via an intermediate that is detectable by differential scanning calorimetry but not CD spectroscopy, consistent with a folded p53tetTD monomer that is equivalent to the p53tet primary dimer. Given its oligomeric stability and resistance against hetero oligomerization, dimerization of p53 constructs incorporating the tetramerization domain may yield functional constructs that may resist exchange with wild-type or mutant forms of p53. PMID- 17113102 TI - Biogenesis of eel liver citrate carrier (CIC): negative charges can substitute for positive charges in the presequence. AB - A family of structurally related carrier proteins mediates the flux of metabolites across the mitochondrial inner membrane. Differently from most other mitochondrial proteins, members of the carrier family are synthesized without an amino-terminal targeting sequence. However, in some mammalian and plant species, representatives were identified that carry a positively charged presequence. To obtain data on a carrier protein from lower vertebrates, we determined the primary structure of eel mitochondrial citrate carrier (CIC) and investigated its import pathway into the target organelle. The protein carries a cleavable presequence of 20 amino acids, including two positively charged residues. The cleavage site is recognized by a magnesium-dependent peptidase in the intermembrane space. The presequence is dispensable both for targeting and translocation, but prior to import into mitochondria, significantly increases the solubility of the precursor protein. This effect is completely retained if the positive charges are exchanged with negative charges. Following this observation, we found that several carrier proteins appear to carry non-cleavable presequences that may similarly act as charged intramolecular chaperones. PMID- 17113103 TI - Biophysical characterisation of the small ankyrin repeat protein myotrophin. AB - The 118 residue protein myotrophin is composed of four ankyrin repeats that stack linearly to form an elongated, predominantly alpha-helical structure. The protein folds via a two-state mechanism at equilibrium. The free energy change of unfolding in water (DeltaG(U-N)(H(2)O)) is 5.8 kcal.mol(-1). The chevron plot reveals that the folding reaction has a broad energy barrier and that it conforms to a two-state mechanism. The rate of folding in water (k(f)(H(2)O)) of 95 s(-1) is several orders of magnitude slower than the value predicted by topological calculations. Proline mutants were used to show that the minor kinetic phases observed for myotrophin arise from heterogeneity of the ground states due to cis trans isomerisation of prolyl as well as non-prolyl peptide bonds. Myotrophin is the first example of a naturally occurring ankyrin repeat protein that conforms to an apparent two-state mechanism at equilibrium and under kinetic conditions, making it highly suitable for high resolution protein folding studies. PMID- 17113104 TI - The crystal structure of the secreted dimeric form of the hemophore HasA reveals a domain swapping with an exchanged heme ligand. AB - To satisfy their iron needs, several Gram-negative bacteria use a heme uptake system involving an extracellular heme-binding protein called hemophore. The function of the hemophore is to acquire free or hemoprotein-bound heme and to transfer it to HasR, its specific outer membrane receptor, by protein-protein interaction. The hemophore HasA secreted by Serratia marcescens, an opportunistic pathogen, was the first to be identified and is now very well characterized. HasA is a monomer that binds one b heme with strong affinity. The heme in HasA is highly exposed to solvent and coordinated by an unusual pair of ligands, a histidine and a tyrosine. Here, we report the identification, the characterization and the X-ray structure of a dimeric form of HasA from S. marcescens: DHasA. We show that both monomeric and dimeric forms are secreted in iron deficient conditions by S. marcescens. The crystal structure of DHasA reveals that it is a domain swapped dimer. The overall structure of each monomeric subunit of DHasA is very similar to that of HasA but formed by parts coming from the two different polypeptide chains, involving one of the heme ligands. Consequently DHasA binds two heme molecules by residues coming from both polypeptide chains. We show here that, while DHasA can bind two heme molecules, it is not able to deliver them to the receptor HasR. However, DHasA can efficiently transfer its heme to the monomeric form that, in turn, delivers it to HasR. We assume that DHasA can function as a heme reservoir in the hemophore system. PMID- 17113105 TI - Lack of strand-specific repair of UV-induced DNA lesions in three genes of the archaeon Sulfolobus solfataricus. AB - In all organisms, specialized systems are devoted to repair of DNA lesions induced by exposure to UV light. In both Eucarya and Bacteria, UV-induced pyrimidine dimers in the transcribed strand of active genes are repaired at a faster rate compared to the non-transcribed strand and the rest of the genome. Preferential repair of transcribed strands requires the Transcription-Repair Coupling Factor in Escherichia coli and the CSA and CSB proteins in humans. These factors are needed for coupling of transcription to nucleotide excision repair (NER), a major pathway for repair of UV-induced lesions. Whereas transcription coupled NER (TC-NER) is an evolutionary conserved process, not all active genes show preferential repair of transcribed strands. The existence of a NER pathway in the Archaea has not been demonstrated directly, yet it is suggested by the presence and properties of homologues of NER nucleases and helicases. However, none of the proteins responsible for the lesion recognition steps or for TC-NER has been found in archaeal genomes. Moreover, the kinetics of gene or strand specific repair has never been investigated in any organism of this domain. We have analysed the kinetics of repair of UV-induced DNA damage in the transcribed and non-transcribed strands of three genes of the hyperthermophilic archaeon Sulfolobus solfataricus. We found that in all three genes the two strands are repaired with the same efficiency with each other and with the genome in general, thus providing no evidence of strand bias or transcription coupling of the repair process in the genes analysed. Further studies will be required to test the existence of a transcription-coupled repair pathway in other archaeal genes and to elucidate the mechanism of UV lesion recognition and repair in Archaea. PMID- 17113106 TI - Functionally important residues in the peptidyl-prolyl isomerase Pin1 revealed by unigenic evolution. AB - Pin1 is a phosphorylation-dependent member of the parvulin family of peptidyl prolyl isomerases exhibiting functional conservation between yeast and man. To perform an unbiased analysis of the regions of Pin1 essential for its functions, we generated libraries of randomly mutated forms of the human Pin1 cDNA and identified functional Pin1 alleles by their ability to complement the Pin1 homolog Ess1 in Saccharomyces cerevisiae. We isolated an extensive collection of functional mutant Pin1 clones harboring a total of 356 amino acid substitutions. Surprisingly, many residues previously thought to be critical in Pin1 were found to be altered in this collection of functional mutants. In fact, only 17 residues were completely conserved in these mutants and in Pin1 sequences from other eukaryotic organisms, with only two of these conserved residues located within the WW domain of Pin1. Examination of invariant residues provided new insights regarding a phosphate-binding loop that distinguishes a phosphorylation-dependent peptidyl-prolyl isomerase such as Pin1 from other parvulins. In addition, these studies led to an investigation of residues involved in catalysis including C113 that was previously implicated as the catalytic nucleophile. We demonstrate that substitution of C113 with D does not compromise Pin1 function in vivo nor does this substitution abolish catalytic activity in purified recombinant Pin1. These findings are consistent with the prospect that the function of residue 113 may not be that of a nucleophile, thus raising questions about the model of nucleophilic catalysis. Accordingly, an alternative catalytic mechanism for Pin1 is postulated. PMID- 17113107 TI - Antineuronal antibodies in a group of children with obsessive-compulsive disorder and Tourette syndrome. AB - An autoimmune hypothesis has been suggested for early onset obsessive-compulsive disorder and Tourette syndrome. The term: Paediatric autoimmune neuropsychiatric disorders associated with streptococcal infection (PANDAS) has been proposed as an aetiological subtype of OCD and TS, related to a Group A beta haemolytic streptococcal (GABHS) infection that triggers an autoimmune response. Antineural antibodies have been studied and found in the sera of some patients with these disorders, and they are thought to cross-react with streptococcal and basal ganglia antigens. The present study included 32 prepubertal-onset OCD patients, 21 with TS diagnosis (some of them meeting criteria for PANDAS) and 19 normal children, all aged between 9 and 17 years. Antibodies were assayed by immunohistochemistry and immunoblot. Special attention was paid to the methodology and a high serum dilution was used to minimize non-specific binding. No anti-basal ganglia antibodies were detected by immunohistochemistry in any of the samples. Two proteins, with approximate molecular weights of 86 kDa and 55 kDa, were found in sera from 7 patients. Though the study supports the hypothesis of an autoimmune process underlying OCD or TS in some patients, further research is needed. PMID- 17113108 TI - Modulation of the reaction rate of regulating protein induces large morphological and motional change of amoebic cell. AB - Morphologies of moving amoebae are categorized into two types. One is the "neutrophil" type in which the long axis of cell roughly coincides with its moving direction. This type of cell extends a leading edge at the front and retracts a narrow tail at the rear, whose shape has been often drawn as a typical amoeba in textbooks. The other one is the "keratocyte" type with widespread lamellipodia along the front side arc. Short axis of cell in this type roughly coincides with its moving direction. In order to understand what kind of molecular feature causes conversion between two types of morphologies, and how two typical morphologies are maintained, a mathematical model of amoebic cells is developed. This model describes movement of cell and intracellular reactions of activator, inhibitor and actin filaments in a unified way. It is found that the producing rate of activator is a key factor of conversion between two types. This model also explains the observed data that the keratocyte type cells tend to rapidly move along a straight line. The neutrophil type cells move along a straight line when the moving velocity is small, but they show fluctuated motions deviating from a line when they move as fast as the keratocyte type cells. Efficient energy consumption in the neutrophil type cells is predicted. PMID- 17113109 TI - A field experimental study on recolonization and succession of subtidal macrobenthic community in sediment contaminated with industrial wastes. AB - A field experiment was carried out in Hong Kong to study the patterns of recolonization and succession of subtidal macrobenthos in defaunated sediment contaminated with industrial wastes and to determine the time required for benthic recovery in the industrial-contaminated sediment. A total of 50 species was found with an average of 172 animals/tray and 24 species/tray recorded one month after deployment. Initial colonizers were predominantly polychaetes (96 animals/tray, accounting for 55.7%) and gastropods (47 animals/tray, accounting for 27.2%). Abundance of macrobenthos increased quickly to a peak (505 animals/tray) after four months, declined afterwards, and increased again till the end of the experiment. Species number peaked (57 species/tray) in the same month as abundance did, and gradually declined thereafter. Abundance, species number and diversity were significantly lower in the industrial-contaminated sediment as compared to the controls during the early successional stages, indicating the harmful effects of industrial wastes on recolonization and succession of macrobenthos. Although no significant differences in community parameters between the industrial-contaminated and the control sediments were found after eleven months, significant difference in species composition still existed after fourteen months, showing a relatively long-term impact of industrial wastes on macrobenthic community structure. PMID- 17113110 TI - Call for pellets! International Pellet Watch global monitoring of POPs using beached plastic resin pellets. PMID- 17113111 TI - Pharmacological and computational analysis of alpha-subunit preferential GABA(A) positive allosteric modulators on the rat septo-hippocampal activity. AB - Clinically most active anxiolytic drugs are positive allosteric modulators (PAMs) of GABA(A) receptors, represented by benzodiazepine compounds. Due to their non selective profile, however, they potently modulate several sup-type specific GABA(A) receptors, contributing to their broad-range side effects. Based on observations in genetically altered mice, however, it has been proposed that anxiolytic action of benzodiazepines is predominantly mediated by GABA(A) alpha2/3 subunit-containing receptors. In the present study we analyzed the actions of the preferential GABA(A) alpha1 and alpha2/3 PAMs, zolpidem and L 838417, respectively on hippocampal EEG and medial septum neuronal activity in anesthetized rats. In parallel, a computational model was constructed to model pharmacological actions of these compounds on the septo-hippocampal circuitry. The present results demonstrated that zolpidem inhibited theta oscillation both in the hippocampus and septum, and profoundly inhibited firing activity of septal neurons. L-838417 also inhibited hippocampal and septal theta oscillation, however, it did not significantly alter firing rate activity of septal neurons. Our computational model showed that cessation of periodic firing of hippocampo septal neurons, representing absence of hippocampal theta activity, disrupted oscillation of septal units, without altering their overall firing activity, similar to changes observed in our in vivo experiments following administration of L-838417. Understanding the correlation between changes in septo-hippocampal activity and actions of selective modulators of GABA(A) subtype receptor modulators would further advance design of anxiolytic drugs. PMID- 17113112 TI - Effects of (S)-3,4-DCPG, an mGlu8 receptor agonist, on inflammatory and neuropathic pain in mice. AB - In this study, the effect of (S)-3,4-dicarboxyphenylglycine (DCPG), a selective mGlu8 receptor agonist, has been investigated in inflammatory and neuropathic pain models in order to elucidate the role of mGlu8 receptor in modulating pain perception. Inflammatory pain was induced by the peripheral injection of formalin or carrageenan in awake mice. Systemic administration of (S)-3,4-DCPG, performed 15 min before formalin, decreased both early and delayed nociceptive responses of the formalin test. When this treatment was carried out 15 min after the peripheral injection of formalin it still reduced the late hyperalgesic phase. Similarly, systemic (S)-3,4-DCPG reduced carrageenan-induced thermal hyperalgesia and mechanical allodynia when administered 15 min before carrageenan, but no effect on pain behaviour was observed when (S)-3,4-DCPG was given after the development of carrageenan-induced inflammatory pain. When microinjected into the lateral PAG (RS)-alpha-methylserine-O-phoshate (MSOP), a group III receptor antagonist, antagonised the analgesic effect induced by systemic administration of (S)-3,4-DCPG in both of the inflammatory pain models. Intra-lateral PAG (S) 3,4-DCPG reduced pain behaviour when administered 10 min before formalin or carrageenan; both the effects were blocked by intra-lateral PAG MSOP. (S)-3,4 DCPG was ineffective in alleviating thermal hyperalgesia and mechanical allodynia 7 days after the chronic constriction injury of the sciatic nerve, whereas it proved effective 3 days after surgery. Taken together these results suggest that stimulation of mGlu8 receptors relieve formalin and carrageenan-induced hyperalgesia in inflammatory pain, whereas it would seem less effective in established inflammatory or neuropathic pain. PMID- 17113113 TI - More accurate sound localization induced by short-term light deprivation. AB - Crossmodal reorganization processes in the brain are mainly associated with early blindness, on the assumption that recruitment of genuine visual areas, such as primary visual cortex, for non-visual functions results in superior auditory and tactile performance of blind, compared to sighted, humans. This study shows that in sighted subjects the accuracy of sound localization, measured by a task of head pointing to acoustic targets, is reversibly increased after short-term light deprivation of 90 min. However, only the systematic deviations from target positions (constant error) were reduced after light deprivation, while the general precision of head pointing remained unchanged. Return to pre-deprivation values was observed after 180 min of re-exposure to light. The post-deprivation change was similar, though less in magnitude, to the effect of blindness that was demonstrated previously. Generally, these findings indicate that auditory-visual crossmodal plasticity can be quite rapidly initiated by deprivation of the visual cortex from visual input. It seems possible that visual deprivation has an influence on neuronal circuits, that are involved in processing of auditory information in visual brain areas of normal sighted humans. Since exclusively the constant error in sound localization, not general performance, was changed, the present effect of visual deprivation may, however, not be attributable to reorganization processes in the sense of a compensation for the absence of vision. It is more likely that the observed change in accuracy was specifically induced by the absence of visual calibration of the neural representation of auditory space during light deprivation. PMID- 17113114 TI - Mental representation of space: insights from an oblique distribution of hallucinations. AB - Three-dimensional spatial distributions of hypnagogic and hypnopompic hallucinations associated with sleep paralysis were used to investigate the internal representation of space. Left-right asymmetries in human preferences and abilities are well established. Parallel effects are also observed as lower-upper asymmetries. These parallels could reflect common underlying mechanisms or additive effects of independently evolved horizontal and vertical asymmetries. This study adds to the growing literature on multidimensional spatial biases in a context free from the influence of task-related factors. We present evidence of an oblique bias in the projection of both sensory and motor hallucinations toward lower-left and especially upper-right external space exceeding that accounted for by an additive model of separate horizontal and vertical biases. These observations are consistent with theories regarding a systematic functional relation of hemispheric with ventral and dorsal cerebral organization. PMID- 17113115 TI - Local and global auditory processing: behavioral and ERP evidence. AB - Differential processing of local and global visual features is well established. Global precedence effects, differences in event-related potentials (ERPs) elicited when attention is focused on local versus global levels, and hemispheric specialization for local and global features all indicate that relative scale of detail is an important distinction in visual processing. Observing analogous differential processing of local and global auditory information would suggest that scale of detail is a general organizational principle of the brain. However, to date the research on auditory local and global processing has primarily focused on music perception or on the perceptual analysis of relatively higher and lower frequencies. The study described here suggests that temporal aspects of auditory stimuli better capture the local-global distinction. By combining short (40 ms) frequency modulated tones in series to create global auditory patterns (500 ms), we independently varied whether pitch increased or decreased over short time spans (local) and longer time spans (global). Accuracy and reaction time measures revealed better performance for global judgments and asymmetric interference that were modulated by amount of pitch change. ERPs recorded while participants listened to identical sounds and indicated the direction of pitch change at the local or global levels provided evidence for differential processing similar to that found in ERP studies employing hierarchical visual stimuli. ERP measures failed to provide evidence for lateralization of local and global auditory perception, but differences in distributions suggest preferential processing in more ventral and dorsal areas respectively. PMID- 17113116 TI - Pre-partum monensin supplementation improves body reserves at calving and milk yield in Holstein cows dried-off with low body condition score. AB - The objective of this study was to determine the effect of a monensin controlled release capsule administered intraruminally at drying-off on body condition score (BCS) at calving, milk yield, fertility and concentration of energy-related blood metabolites in Holstein cows dried-off with low BCS (< or = 3.0, scale 1 to 5 with a 0.25 point of increment). Between July and August, 2001, 220 cows from parity 2 or more and dried-off 50-70 days before expected parturition, with a BCS < or = 3.0 were randomly assigned to either a treatment group (n=110; oral capsule of monensin releasing 335 mg/day for 95 days) or a control group (no capsule, n=110). At assignment, on day 21 before expected parturition, at calving, and at 7, 14, and 21 days in milk a blood sample was obtained from a random sub sample of 10 cows per group. Effects of monensin on serum NEFA, BHBA and glucose were measured. Milk yield, milk fat and protein content (%) at DHIA test days during the entire lactation, 305 ME milk production and reproductive responses were compared. Monensin significantly improved BCS at calving, increased milk yield at test days 4 and 8, decreased the percentage of milk protein, did not change the percentage of milk fat, and decreased NEFA and BHBA during the post-partum period. PMID- 17113117 TI - Serological screening and toxoplasmosis exposure factors among pregnant women in the Democratic Republic of Sao Tome and Principe. AB - The seroprevalence of Toxoplasma gondii infection among pregnant women in the Democratic Republic of Sao Tome and Principe (DRSTP) from November 2003 to March 2004 was determined by detection of serum anti-T. gondii antibodies. A short questionnaire interview for pregnant women was performed to investigate risk factors associated with T. gondii infection, including consumption of raw meat or unwashed vegetables, drinking unboiled water and keeping pets (cats and dogs). The overall seroprevalence of T. gondii infection was high (75.2%; 375/499). The older age group of > or =35 years had a significantly higher seroprevalence (85.7%; 54/63) than that of the younger age group of 15-25 years (70.4%; 178/253) (odds ratio 2.5, 95% CI 1.2-5.4; P=0.01). No significant difference in the seroprevalence of T. gondii infection was found between the pregnant women with and without exposure to the risk factors studied. However, among pregnant women with high antibody titers of > or =1:1024, it seemed likely that continual contact with pets and consumption of oocyst-contaminated water or raw unwashed vegetables rather than tissue cysts in meat was the primary route of infection. The incidence of congenital toxoplasmosis in unborn babies should be intensively monitored in the DRSTP. PMID- 17113118 TI - Effect of sphingomyelin and cholesterol on the interaction of St II with lipidic interfaces. AB - Sticholysin II (St II) is a cytolysin produced by the sea anemone Stichodactyla helianthus, characterized by forming oligomeric pores in natural and artificial membranes. In the present work the influence of the membrane lipidic components sphingomyelin (SM) and cholesterol (Cho) on binding and functional activity of St II, was evaluated using ELISA, lipid monolayers and liposomes. The aim of this work was to establish the promoting role of Cho and SM, both in St II binding and pore formation efficiency. In general the association (evaluated by ELISA and incorporation to phospholipid monolayers) of St II to lipids mixtures was better than to any one of the single components. Regarding the unique role of SM, it was found that, albeit inefficiently, St II binds to phosphatidylcholine (PC):Cho monolayers and liposomes, and is able to form active pores in these bilayers. The results in monolayers and liposomes show that the presence of SM and large amounts of Cho leads to the highest values of critical pressure and rate of association to monolayers, the most favorable interaction with liposomes, and the fastest rate of pore formation, in spite of the rigidity of the layers as suggested by the high generalized polarization (GP) of Laurdan incorporated to liposomes and FTIR data. Taken together, the present results show that the joint presence of SM and Cho, both in binary and ternary (PC containing) mixtures provide conditions particularly suitable for St II binding and function. We suggest that microdomains present in the bilayers could be important for toxin membrane association. PMID- 17113119 TI - Optimization of ciguatoxin extraction method from blood for Pacific ciguatoxin (P CTX-1). AB - Ciguatera diagnosis relies on clinical observations associated with a recent consumption of fish. Although needed, direct confirmation of exposure in subjects showing ciguatera disease symptoms is currently unavailable. We previously reported that ciguatoxins were measurable in the blood of mice exposed to extracts of Pacific ciguatoxins isolated from Gambierdiscus polynesiensis, and of Indian Ocean or Caribbean Sea ciguatoxins, isolated from fish. Although highly efficient for extracting spiked purified Caribbean-CTX-1, the methanolic extraction method previously described is found here to yield only 6% recovery of spiked Pacific-CTX-1 (P-CTX-1). We report in this short communication a substantially modified method for ciguatoxin extraction from both dried and fresh blood. With this method, toxin measurement is directly accomplished in acetonitrile deproteinated whole fresh blood or phosphate buffer solution (PBS) eluted dried blood using the N2A cell-based assay. Spike studies using increasing concentrations of purified ciguatoxins reveal linear (r2 above 0.87 for all toxins) and overall efficient toxin recoveries (62%, 96%, and 96% from fresh blood and 75%, 90%, and 74% from dried blood, for C-CTX-1, P-CTX-3C, and P-CTX-1, respectively). Comparative blood matrix analysis for P-CTX-1 recovery shows increased recovery of ciguatoxin activity from whole fresh blood than from dried blood, greater by 20% in P-CTX-1 spiked mice blood and by over 85% in P-CTX-1 exposed mouse blood. In conclusion, both Caribbean and Pacific ciguatoxins can be readily extracted from blood using this modified method; however, in the case of P-CTX-1 we find that fresh blood is optimal. PMID- 17113120 TI - Murine AIDS requires CD154/CD40L expression by the CD4 T cells that mediate retrovirus-induced disease: Is CD4 T cell receptor ligation needed? AB - LP-BM5, a retroviral isolate, induces a disease featuring an acquired immunodeficiency syndrome termed murine AIDS (MAIDS). Many of the features of the LP-BM5-initiated disease are shared with HIV/AIDS. Our lab has shown that the interaction of B and CD4 T cells that is central to MAIDS pathogenesis requires ligation of CD40 on B cells by CD154 on CD4 T cells. Despite this strict requirement for CD154 expression, whether CD4 T cell receptor (TCR) occupancy is essential for the induction of MAIDS is unknown. To block TCR engagement, Tg mouse strains with monoclonal TCR of irrelevant peptide/MHC specificities, all on MAIDS-susceptible genetic backgrounds, were tested: the study of a panel of TCR Tg CD4 T cells controlled for the possibility of serendipitous crossreactive recognition of virus-associated or induced-self peptide, or superantigen, MHC complexes by a given TCR. The results argue that TCR engagement is not necessary for the induction of MAIDS. PMID- 17113121 TI - Bone-marrow-derived mesenchymal stem cells as a target for cytomegalovirus infection: implications for hematopoiesis, self-renewal and differentiation potential. AB - Mesenchymal stem cells (MSCs) in bone marrow (BM) regulate the differentiation and proliferation of adjacent hematopoietic precursor cells and contribute to the regeneration of mesenchymal tissues, including bone, cartilage, fat and connective tissue. BM is an important site for the pathogenesis of human cytomegalovirus (HCMV) where the virus establishes latency in hematopoietic progenitors and can transmit after reactivation to neighboring cells. Here we demonstrate that BM-MSCs are permissive to productive HCMV infection, and that HCMV alters the function of MSCs: (i) by changing the repertoire of cell surface molecules in BM-MSCs, HCMV modifies the pattern of interaction between BM-MSCs and hematopoietic cells; (ii) HCMV infection of BM-MSCs undergoing adipogenic or osteogenic differentiation impaired the process of differentiation. Our results suggest that by altering BM-MSC biology, HCMV may contribute to the development of various diseases. PMID- 17113122 TI - Quantitative analysis of herpes simplex virus type 1-specific memory B cells generated by different routes of infection. AB - We compared the herpes simplex virus type 1 (HSV)-specific memory B cell (MBC) populations generated by footpad and intranasal infection in mice. Both routes of infection generated transient antibody-secreting cell responses in the draining lymph nodes and spleen, and sustained circulating IgG. HSV-specific IgG MBCs, analyzed by limiting dilution assay approximately 8 weeks after infection, were distributed in a range of lymph nodes and in the spleen and Peyer's patches. Overall, the route of infection had little effect on the MBC frequency in each anatomical location. Interestingly, after both routes of infection there was a trend towards preferential MBC accumulation in the mediastinal lymph node. Intravaginal challenge of mice primed by footpad or intranasal infection generated similar secondary IgG responses. Our findings indicate that the widespread dispersion of MBCs to lymphoid tissues throughout the body is largely independent of the route of infection, but may be influenced by tissue-specific factors. PMID- 17113123 TI - Quantitative evaluation of bacteria released by bathers in a marine water. AB - Enterococci, a common fecal indicator, and Staphylococcus aureus, a common skin pathogen, can be shed by bathers affecting the quality of recreational waters and resulting in possible human health impacts. Due to limited information available concerning human shedding of these microbes, this study focused on estimating the amounts of enterococci and S. aureus shed by bathers directly off their skin and indirectly via sand adhered to skin. Two sets of experiments were conducted at a marine beach located in Miami-Dade County, Florida. The first study, referred to as the "large pool" study, involved 10 volunteers who immersed their bodies in 4700L during four 15min cycles with exposure to beach sand in cycles 3 and 4. The "small pool" study involved 10 volunteers who were exposed to beach sand for 30min before they individually entered a small tub. After each individual was rinsed with off-shore marine water, sand and rinse water were collected and analyzed for enterococci. Results from the "large pool" study showed that bathers shed concentrations of enterococci and S. aureus on the order of 6x10(5) and 6x10(6) colony forming units (CFU) per person in the first 15min exposure period, respectively. Significant reductions in the bacteria shed per bather (50% reductions for S. aureus and 40% for enterococci) were observed in the subsequent bathing cycles. The "small pool" study results indicated that the enterococci contribution from sand adhered to skin was small (about 2% of the total) in comparison with the amount shed directly from the bodies of the volunteers. Results indicated that bathers transport significant amounts of enterococci and S. aureus to the water column, and thus human microbial bathing load should be considered as a non-point source when designing recreational water quality models. PMID- 17113124 TI - The use of oxyhalogen in photocatalytic reaction to remove o-chloroaniline in TiO2 dispersion. AB - Photodecay of o-chloroaniline (o-ClA) in various combinations of UV sources, TiO2, and oxyhalogens was investigated. To improve the conventional photocatalytic process by using UV/TiO2, the addition of oxyhalogens (ClO3(-), BrO3(-) and IO3(-)) into UV/TiO2 system was studied and the effect in such addition is very encouraging for all the selected additives. Oxyhalogens are capable of deferring the electron-hole recombination of TiO2 which significantly improved its catalytic performance. The presence of IO3(-) in UV/TiO2 resulted in the fastest o-ClA decay among three oxhalogens at the same dosage. The decay of o ClA in UV/TiO2/oxyhalogen process is characterized by a two-stage pseudo kinetics, where a faster initial decay was followed by a retardation state. A mathematics model was successfully established for the prediction of the two stage decay of o-ClA in UV/TiO2/IO3(-) with any designed [IO3(-)] concentration. PMID- 17113125 TI - Quantitative structure-activity relationships for toxicity and genotoxicity of halogenated aliphatic compounds: wing spot test of Drosophila melanogaster. AB - Halogenated aliphatic compounds were evaluated for toxic and genotoxic effects in the somatic mutation and recombination test employing Drosophila melanogaster. The tested chemicals included chlorinated, brominated and iodinated; mono-, di- and tri-substituted; saturated and unsaturated alkanes: 1,2-dibromoethane, 1 bromo-2-chloroethane, 1-iodopropane, 2,3-dichloropropene, 3-bromo-1-propene, epibromohydrin, 2-iodobutane, 3-chloro-2-methylpropene, 1,2,3-trichloropropane, 1,2-dichloroethane, 1,2-dichlorobutane, 1-chloro-2-methylpropane, 1,3 dichloropropane, 1,2-dichloropropane, 2-chloroethymethylether, 1-bromo-2 methylpropane and 1-chloropentane. N-methyl-N-nitrosourea served as the positive and distilled water as the negative control. The set of chemicals for the toxicological testing was selected by the use of statistical experiment design. Group of unsaturated aliphatic hydrocarbons were generally more toxic than saturated analogues. The genotoxic effect was observed with 14 compounds in the wing spot test, while 3 substances did not show any genotoxicity by using the wing spot test at 50% lethal concentration. The highest number of wing spots was observed in genotoxicity assay with 1-bromo-2-chloroethane, 1,2-dichloroethane, 1,2-dibromoethane and 1-iodopropane. Nucleophilic superdelocalizability calculated by quantum mechanics appears to be a good parameter for prediction of both toxicity and genotoxicity effects of halogenated aliphatic compounds. PMID- 17113126 TI - The toxic effects of neem extract and azadirachtin on the brown planthopper, Nilaparvata lugens (Stal) (BPH) (Homoptera: Delphacidae). AB - Extracts of neem (Azadirachta indica A. Juss) are used in the developing world for many purposes including management of agricultural insect pests. The effects of different neem extracts (aqueous (NSKEaq), ethanol (NSKEeth) and hexane (NSKEhex)) on mortality, survival and weight of the brown planthopper, Nilaparvata lugens (Stal) (BPH) (Homoptera: Delphacidae) third and fourth nymphal instars were investigated. When fed rice plants treated with neem derivatives in bioassays, the survival of BPH nymphs is affected. Comparisons were made with the pure neem limonoid, azadirachtin (AZA) to ascertain its role as a compound responsible for these effects. AZA was most potent in all experiments and produced almost 100% nymphal mortality at 0.5 ppm and higher concentrations. When higher concentrations were applied, the effects appeared shortly after treatment and mortality was higher. Many insects died after remaining inactive for several days or during prolonged moulting. At lower concentrations, if moulting was achieved, disturbed growth and abnormalities were then likely to occur in the moulting process. Nymphs that were chronically exposed to neem extract showed a reduction in weight (45-60%). The results clearly indicate the simple NSKE (aqueous, ethanolic or both), containing low concentrations of AZA, can be used effectively to inhibit the growth and survival of BPH. PMID- 17113127 TI - Partitioning and photodegradation of ciprofloxacin in aqueous systems in the presence of organic matter. AB - Ciprofloxacin is an extensively used antibiotic that has been reported to occur in surface water. Previous studies have indicated that ciprofloxacin photodegrades and sorbs to particulate organic material within aquatic systems. The first objective of the current study was to evaluate the influence of organic material on photodegradation rates of ciprofloxacin. Using a bench top experimental design, ciprofloxacin was added to experimental chambers that contained only water or water and fine particulate organic matter (FPOM) followed by exposure to ultraviolet light. Sorption to FPOM was rapid, reducing the amount of ciprofloxacin that was available for photodegradation. Thus, the presence of FPOM initially decreased the ciprofloxacin concentration in the aqueous compartment. However by the end of the 16 h test, 42% of the ciprofloxacin was recovered from the test system with FPOM present, while only 2% of the ciprofloxacin was recovered in systems that did not contain FPOM. The second objective of this study was to compare the sorption coefficients for ciprofloxacin between two types of organic material: FPOM, classified as amphipod processed leaves, and coarse particulate organic matter (CPOM), represented by intact leaf disks. Sorption to FPOM (log Kd of 4.54+/-0.09 l kg(-1)) was 1.6 orders of magnitude greater than sorption to CPOM (log Kd of 2.92+/-0.10 l kg( 1)) potentially resulting in differential toxicity among similar organisms that occupy these different niches and leading to different estimates of environmental fate and effects. PMID- 17113128 TI - Distribution and accumulation of heavy metals in the sediments of Kaohsiung Harbor, Taiwan. AB - The distribution, enrichment, and accumulation of heavy metals in the sediments, especially those at the vicinity of tributary estuaries of Kaohsiung Harbor, Taiwan were investigated. Sediment samples from six locations in the Kaohsiung Harbor were collected quarterly in the period from 2002 to 2005 and characterized for metal content (e.g., Hg, Pb, Cd, Cr, Cu, Zn and Al), water content, organic matter, total nitrogen, total phosphorous, total grease, and grain size. Results showed that metal concentrations varied from 0.58 mg kg(-1) for Cd to 596 mg kg( 1) for Zn. Metal concentrations at the vicinity of river mouths were higher than those at other locations. All heavy metals studied, except Cr, had relatively high enrichment factors and geo-accumulation indices in the estuaries. Moreover, metal concentrations correlated closely to the physical-chemical properties of the sediments, which strongly suggested the influence of industrial and municipal wastewaters discharged from the neighboring industrial parks and river basins. Results would help develop strategies for pollution control and sediment remediation of Kaohsiung Harbor. PMID- 17113129 TI - Radiolysis of aqueous 4-nitrophenol solution with Al2O3 or TiO2 nanoparticles. AB - Aqueous 4-nitrophenol solutions containing TiO2 or Al2O3 nanoparticles were irradiated with electron beam. 4-Nitrophenol was decomposed by the ionizing radiation process in the absence of the nanoparticles. The addition of TiO2 or Al2O3 (2 g l(-1)) before irradiation improved the removal of 4-nitrophenol, total organic carbon (TOC) but also nitrogen (TN). To identify the origin of the loss (catalysis or simply adsorption), TiO2 or Al2O3 nanoparticles were added after irradiation. Experiments show that the effect of the presence of TiO2 or Al2O3 during irradiation is just due to adsorption. PMID- 17113130 TI - Application of advanced technologies in ageing research. AB - Several technologies that emerged in the post-genomic era have been particularly useful in dissecting the molecular mechanisms of complex biological processes through the systems approach. Here, we review how three of these technologies, namely transcriptional profiling, large-scale RNA interference (RNAi) and genome wide location analysis of protein-DNA interactions, have been used in the study of ageing in metazoans. We also highlight recent developments of these three technologies and how these developments are applicable to ageing research. PMID- 17113131 TI - Studies on commuters' exposure to BTEX in passenger cars in Kolkata, India. AB - Commuters' exposure to volatile organic compounds (VOCs) especially BTEX travelling in passenger cars in Kolkata, India were quantified in Phase I (2001 2002) and Phase II (2003-2004). Monitoring was made inside and in the immediate outside of passenger cars fitted with and without catalytic converters using different types of fuels, along two congested urban routes. During Phase I of the study, the benzene content in gasoline was 5% and the mean concentration of in vehicle benzene in cars without catalytic converter was found to be as high as 721.2 microg/m3. In Phase II when the benzene content was reduced to <3% and with modified engine type, the mean in-vehicle benzene concentration was reduced to 112.4 microg/m3. The in-vehicle concentration varied with engine type and age of the vehicle. Roadside ambient mean concentration of benzene was 214.8 microg/m3 and 30.8 microg/m3 in Phase I and Phase II respectively. PMID- 17113132 TI - Acetylcholinesterase activity in seabirds affected by the Prestige oil spill on the Galician coast (NW Spain). AB - In November 2002, the tanker Prestige broke in two and sank at the bottom of the ocean spilling about 70,000 t of fuel oil, which reached the coast of Galicia. It was considered the largest spill in maritime history, greatly affecting marine and related avian species. The spilled fuel oil contained high concentrations of polycyclic aromatic hydrocarbons (PAHs). Many species were affected and were found dead, although ongoing research is still being carried out on the sublethal effects. In this sense, little is known about the action of PAHs on Cholinesterase activity in seabirds. Consequently, the purpose of this study was to provide more information on the neurotoxicity of fuel oil on the seabirds most affected by the Prestige accident: common guillemot, Atlantic puffin and razorbill. On the other hand, data on normal values of acetylcholinesterase (AChE) activity were obtained to supply non-exposed values in seabirds. The oil spill produced a clear inhibitory effect on brain AChE activity in common guillemot (16%, p< or =0.01) and razorbill (22%, p< or =0.01), but not in Atlantic puffin (4%). Physiological levels of brain AChE, expressed in nmol acetylcholine hydrolysed min(-1) mg(-1) protein were similar in non-exposed common guillemot (388.6+/-95.0) and Atlantic puffin (474.0+/-60.7), however, razorbill values were higher (644.6+/-66.9). PMID- 17113133 TI - Targeting low-arsenic aquifers in Matlab Upazila, Southeastern Bangladesh. AB - Groundwater with high concentration of geogenic arsenic (As) occurs extensively in the Holocene alluvial aquifers of Bangladesh. Local drillers in Matlab Upazilla are constructing deeper tubewells than in the recent past, primarily because of low concentrations of dissolved Fe and As. Locally a thick layer of black to grey sediments overlies an oxidised unit of yellowish-grey to reddish brown sediments. The correlation between the colour of both units and the groundwater redox conditions was investigated to provide an easy tool for targeting low-arsenic groundwater. Based on the sediment colour at the screen depths described by local drillers, 40 domestic shallow tubewells were selected for water sampling. Four colours were used to describe the sediments: black, white, off-white (buff) and red. Generally, the groundwater was anoxic and the As concentrations ranged from less than 5.2 to 355 microg/L. Water derived from the black sediment is characterized by relatively higher concentrations of dissolved NH(4)(+), DOC, Fe, P, As and by low Mn and SO(4)(2-) concentrations. The off white and red sediments had high concentration of Mn and low NH(4)(+), DOC, Fe, P and As concentrations. The water abstracted from the black sediments indicated the most reducing environment, followed by white, off-white and red respectively. Three boreholes verified the driller's perception of the subsurface lithologic conditions. Discrepancies between the driller's and the research team description of the sediment colours were insignificant. This study shows that sediment colour is a reliable indicator of high and low-As concentrations and can be used by local drillers to target low-arsenic groundwater. PMID- 17113134 TI - Melano-macrophage centres and endocytic cells in kidney and spleen of pearl gouramy and platyfish (Anabantidae, Poeciliidae: Teleostei). AB - The structure and putative cellular content of melano-macrophage centres (MMCs) and single macrophages in kidney and spleen of two teleosts, pearl gouramy Trichogaster leeri (Bleeker) and platyfish Xiphophorus maculatus (Gunther), are described and compared. The MMCs were confined to the haemopoetic tissue in both species, and were markedly more prominent in pearl gouramy than in platyfish. They were rich in iron-compounds, probably haemosiderin, in the former species, whereas they contained mainly iron-free pigments, probably lipofuscin, in the latter species. We suggest that the kidney and spleen MMCs are involved in the regular storage, relocation and recycling of iron-compounds of effete or damaged red blood cells in healthy pearl gouramies, whereas they function more or less as a deposit for undegradable and potentially harmful iron-free cell debris in the corresponding organs in healthy platies. Numerous single macrophages took up much intraperitoneally injected horse ferritin in the kidney and spleen haemopoetic tissue in platies, whereas these cells contain much iron-containing pigment, probably haemosiderin, in the control pearl gouramies. We suggest that these cells are able to play a role in cleansing the circulation of foreign macromolecules and particles in platies, whereas they are mainly involved in the regular uptake of scavenger iron-containing compounds in pearl gouramies. PMID- 17113135 TI - A two-generation chronic mixture toxicity study of Clophen A60 and diethyl phthalate on histology of adrenal cortex and thyroid of rats. AB - This study was undertaken to observe the type of interaction that exists between polychlorinated biphenyls (Clophen A60) and diethyl phthalate (DEP) on the adrenal and thyroid glands of male and female Wistar rats. Animals were divided into four groups of six animals each, group I male and female rats were fed on a normal diet and water ad libitum. Groups II, III and IV male and female rats were given Clophen A60, DEP, or mixture of Clophen A60 and DEP, respectively, each dissolved in corn oil mixed with the diet at 50mg/kg of the diet/day. One hundred days after treatment, females were mated with males for 10 days. Exposure to the pollutants was continued throughout mating, gestation (21 days) until termination at weaning (21 days), which was 150 days of total treatment period of the parental generation. When the F1-generation pups (six males and six females of each group) were 75-100g in weight, they were treated in a similar manner to the parental generation, again for a period of 150 days, with the dose reduced to 25mg/kg of the diet/day in all treated groups. After 150 days of treatment, animals were sacrificed and histology of the adrenal and thyroid glands was asessed. An antagonistic interactive effect of treatment was seen in male parental and F1-generation rats, while an inhibitory type of interactive effect was observed in female rats. In the zona fasciculata region of the adrenal cortex of treated rats of both generations, vacuolations and degeneration were seen in samples from male animals and intracellular vacuolations in samples from females. A synergistic interactive toxic effect to the thyroid gland was observed in treated parental generation male rats, and mild changes in F1-generation-treated male rats, showing follicular shrinkage, loss of thyroglobulin and fibrosis of the interfollicular epithelium. In females, an antagonistic effect to the thyroid gland was observed in both parental and F1-generation-treated rats, showing similar effects as observed in males. From this study, we can conclude that combined administration of Clophen A60 and DEP shows an enhanced toxic effect on adrenal glands of F1-generation male and female rats, but the effect is much more marked in the thyroid gland of F1-generation male rats, and seen to a lesser extent in F1-generation female rats. PMID- 17113136 TI - Possible role of insulin-like growth factor-II C-peptide on catecholamine release and ultrastructural aspects of chromaffin cells in the adrenal gland of the frog. AB - The present study was undertaken to demonstrate that insulin-like growth factor II C-peptide (IGF-II C-peptide) affects the function of the adrenal gland of Rana ridibunda (Anura, Amphibia) by stimulating chromaffin cells. Previous studies have shown that insulin-like growth factors affect adrenal gland function in mammals. On the basis of these findings, frogs were injected with IGF-II C peptide (2.5 microg/0.2 ml), whereas control animals were injected with Ringer solution (0.2 ml). The adrenal glands were removed at 12 and 48 h after injection and fixed, embedded in paraffin wax and Epon, and examined by immunohistochemistry and transmission electron microscopy to investigate whether there were structural changes and activation of chromaffin cells in the frog adrenal gland. Sections were stained with hematoxylin and eosin for overall tissue analysis and, in parallel, serotonin was localized using the streptavidin biotin complex technique while dopamine beta-hydroxylase was shown by the peroxidase-antiperoxidase-3, 3'-diaminobenzidine tetrachloride method. After injection of IGF-II C-peptide, chromaffin cells released serotonin and synthesized dopamine beta-hydroxylase. The most pronounced effect of IGF-II C peptide on the chromaffin cells was observed at 12h after injection. Our results indicate that there is a possible role of IGF-II C-peptide on chromaffin cell activity enhancing catecholamine release in the adrenal gland of the frog. PMID- 17113137 TI - CA125 expression pattern, prognosis and correlation with serum CA125 in ovarian tumor patients. From The Danish "MALOVA" Ovarian Cancer Study. AB - OBJECTIVES: To determine the CA125 tissue expression levels in borderline and invasive epithelial ovarian tumor tissues. Secondly, to evaluate whether CA125 tissue expression levels correlate with clinico-pathological parameters and serum CA125 levels and finally to investigate the prognostic value of tissue CA125 expression levels in ovarian cancer (OC) patients. METHODS: We designed tissue arrays (TA) and analyzed the CA125 expression in tissues from 778 Danish women with an ovarian tumor. Furthermore, corresponding preoperative blood samples obtained before surgery were collected from 382 women with OC. RESULTS: Significantly more CA125 expression positive tumors (no expression vs. expression) were found in the serous subtype compared to the percentage of positive tumors in mucinous, endometroid and other subtypes for patients both with borderline ovarian tumors and with OC (p<0.00001, p<0.00001). Similarly, a positive significant correlation was found between elevated serum CA125 levels and elevated levels of CA125 tissue expression (N=382 stage I-IV OC, Spearman rho=0.31, p<0.0001) (N=206 stage III OC, Spearman rho=0.30, p<0.0001). We found a significantly shorter survival for stage III/IV OC patients with no CA125 tissue expression compared to stage III/IV OC patients with positive CA125 tissue expression (p=0.0003). CONCLUSION: Our finding that tissue CA125 expression was lacking in late stage primary OC tumor of Danish women with poor survival may be of value in selecting patients as eligible candidates for individually based treatments. PMID- 17113138 TI - Wapl controls the dynamic association of cohesin with chromatin. AB - Cohesin establishes sister-chromatid cohesion from S phase until mitosis or meiosis. To allow chromosome segregation, cohesion has to be dissolved. In vertebrate cells, this process is mediated in part by the protease separase, which destroys a small amount of cohesin, but most cohesin is removed from chromosomes without proteolysis. How this is achieved is poorly understood. Here, we show that the interaction between cohesin and chromatin is controlled by Wapl, a protein implicated in heterochromatin formation and tumorigenesis. Wapl is associated with cohesin throughout the cell cycle, and its depletion blocks cohesin dissociation from chromosomes during the early stages of mitosis and prevents the resolution of sister chromatids until anaphase, which occurs after a delay. Wapl depletion also increases the residence time of cohesin on chromatin in interphase. Our data indicate that Wapl is required to unlock cohesin from a particular state in which it is stably bound to chromatin. PMID- 17113139 TI - Clinical long-term retention of etch-and-rinse and self-etch adhesive systems in non-carious cervical lesions. A 13 years evaluation. AB - OBJECTIVES: The aim of this study was to evaluate the clinical long-term retention to dentin of seven adhesive systems. METHODS: A total of 337 Class V restorations of three three-step etch-and-rinse, one two-step etch-and-rinse and three self-etch adhesive systems were placed in non-carious cervical lesions without intentional enamel involvement. The restorations were evaluated at baseline and then every 6 months during a 13 years follow-up. Dentin bonding efficiency was determined by the percentage of lost restorations. RESULTS: During the 13 years, 275 restorations could be evaluated. The cumulative loss rate at 13 years was 60.3%, with significant different failures rates for the different systems varying between 26.3 and 94.7%. Three materials fulfilled the ADA 18 months full acceptance criteria. Three systems showed already at 18 months or earlier catastrophical debonding rates. The annual failure rates for the three step etch-and-rinse systems were: Allbond 2 4.1%, Clearfil LB 2.0% and Denthesive 7.3%. For the two-step etch-and-rinse Gluma 2000 6.5%, and for the self-etch systems ART 3.2%, Denthesive 2 5.7% and PUB 3 4.5% CONCLUSION: A continuous degradation of the resin-dentin bond was observed for all bonding systems during the follow-up expressed by the increasing loss rates. A wide variation of dentin bonding effectiveness was seen between the systems independent to adhesion strategy. PMID- 17113140 TI - Implications of inadequate parental bonding and peer victimization for adolescent mental health. AB - Previous studies have indicated significant associations between relatively poor mental health of children and both perceived negative parenting and exposure to peer victimization at school. This paper examines their relative contribution to the mental status of adolescent school children. Questionnaires were administered to Australian school children (n=1432) aged 12-16 years. These contained reliable self-report measures of mental health (the GHQ) parental bonding (the PBI) and degree of peer victimization. As predicted, multiple regression analyses indicated that low levels of perceived parental care, high parental control and frequent peer victimization were each significantly and independently associated with relatively poor mental health. Together, they accounted for some 17% and 27% of variance in the mental health status of adolescent boys and girls, respectively. Theoretical and practical implications of these findings are considered. Limitations of the study are also discussed (deleted). PMID- 17113141 TI - Glycosylated trypsin-like proteases from earthworm Eisenia fetida. AB - Although groups of earthworm proteases have been found by several laboratories, it is still unclear how many of the isolated trypsin-like fibrinolytic enzymes are in glycosylated form. Here, eight glycosylated fibrinolytic proteases (EfP-0 1, EfP-0-2, EfP-I-1, EfP-I-2, EfP-II-1, EfP-II-2, EfP-III-1 and EfP-III-2) were isolated from an earthworm species (Eisenia fetida) through a stepwise purification procedure: ammonium sulfate precipitation, affinity chromatography on a Sepharose-4B column coupled with soybean trypsin inhibitor (SBTI), and ionic chromatography with a DEAE-Cellulose-52 column. Among the eight purified trypsin like glyco-proteases, EfP-0-2 and EfP-II-2 were newly isolated isozymes. Glycoprotein staining of the proteases on native-PAGE with a Schiff's reagent (sodium meta-periodate) revealed that the eight proteases were glycoproteins. Measurements of the glycan content with sodium meta-periodate and glycoprotein test reagent showed that these proteases had different carbohydrate contents. Dot blotting assay with ConA suggested the oligosaccharides were composed of mannose residues. PMID- 17113142 TI - Terraced self assembled nano-structures from laminarin. AB - The formation of self assembled nano-structures from the biopolymer laminarin dried onto mica is reported. The observed structures are composed of stacked terraced layers decreasing in size away from the mica surface. The layers display a high degree of dimensional regularity as observed using atomic force microscope imaging (AFM). The width of the layers is linearly dependent upon the number of layers in the structure and decreases with layer number away from the mica substrate. The thickness of the layers is uniform throughout the structure. A pore is contained in the central region of each structure with more than one layer. We postulate that these structures have potential use as templates in microelectronic devices and sensors where the central pore has the potential to immobilise functional materials. PMID- 17113143 TI - Processing and tensile properties of hydroxyapatite-whisker-reinforced polyetheretherketone. AB - Polyetheretherketone (PEEK) was reinforced with 0-50 vol% hydroxyapatite (HA) whiskers using a novel powder processing and compression molding technique which enabled uniform mixing at high whisker content. Texture analysis showed that viscous flow during compression molding produced a preferred orientation of whiskers along the specimen tensile axis. Consequently, the elastic modulus or ultimate tensile strength of HA-whisker-reinforced PEEK was able to be tailored to mimic human cortical bone. PEEK reinforced with 40 and 50 vol% HA whiskers exhibited elastic moduli of 17 and 23 GPa, respectively. Elastic constants were measured using ultrasonic wave propagation and revealed an orthotropic anisotropy also similar to that measured in human cortical bone. PEEK reinforced with 10 and 20 vol% HA whiskers exhibited an ultimate tensile strength of 90 and 75 MPa, respectively. Tensile specimen fracture surfaces showed evidence of brittle failure in both reinforced and un-reinforced PEEK. Whisker pullout was observed with PEEK adhered to HA whiskers, suggesting a relatively strong interface between the PEEK matrix and HA whisker reinforcements. PMID- 17113144 TI - On the biology of saphenous vein grafts fitted with external synthetic sheaths and stents. AB - Autologous saphenous vein is used as a conduit to bypass atherosclerotic lesions in both the coronary artery (coronary artery bypass graft surgery [CABG]) and in femoral arteries (infrainguinal bypass graft surgery [IIBS]). Despite the undoubted success and benefits of the procedures, graft failure occurs in 50% of cases within 10 years after surgery. A principal cause of late vein graft failure is intimal and medial hyperplasia and superimposed atherogenesis. Apart from lipid lowering therapy, no intervention has hitherto proved clinically effective in preventing late vein graft failure which clearly constitutes a major clinical and economic problem that needs to be urgently resolved. However, we have studied the effect of external synthetic stents and sheaths in pig models of vein into artery interposition grafting and found them to have a profound effect on vein graft remodelling and thickening. In this review, therefore, we will summarise the mechanisms underlying vein graft failure and how these stents influence these processes and the possible mechanisms involved as well as the application of these devices in preventing vein graft failure clinically. PMID- 17113145 TI - High expression of survivin and down-regulation of Stat-3 characterize the feto maternal interface in failing murine pregnancies during the implantation period. AB - The materno-fetal interface has for long been considered as an immune privileged biological site and thus understanding the mechanisms underlying fetal survival have been the focus of intense research. In adults, survivin and Stat-3 proteins are involved in tolerance as well as the induction of apoptosis. However, the role of these molecules in pregnancy and development has not been addressed. We have evaluated the expression of survivin and Stat-3 in allogeneic mouse models of low abortions (CBA/J x Balb/c), abortion prone (CBA/J x DBA/2J) and stress triggered abortions from DBA/2J-mated CBA/J mice. We show that survivin is over expressed in abortion-prone mating on gestation day 7.5. This effect was also found in stress-exposed mice, whereas expression was low in normal pregnancy mice. The phosphorylated Stat-3 (p-Stat-3) was down regulated in high abortion mating compared with low abortion mating, CBA/J x Balb/c. The level of apoptosis was similar in the three groups studied. Our results suggest that high expression of survivin and low expression of p-Stat-3 are involved in pregnancy loss in mice. PMID- 17113146 TI - Reduction of maternal adrenal steroids results in increased VEGF protein without increased eNOS in the ovine placenta. AB - Fetal sheep studies have shown that reduced maternal cortisol or aldosterone levels alter placental morphology, with a reduction in placental blood flow. We have now tested the hypothesis that changes in placental morphology with relative adrenal hypoadrenalism are associated with changes in vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS). Four groups of late gestation pregnant ewes with singleton fetuses were studied; controls (intact adrenals), normal cortisol and aldosterone (ewes adrenalectomized and replaced with normal cortisol and aldosterone levels), low cortisol (ewes adrenalectomized and replaced with low cortisol levels), and low aldosterone (ewes adrenalectomized and replaced with low aldosterone levels). The placenta was categorized into A, B, C or D type placentomes. There were significantly more B and C type placentomes in the adrenalectomized groups than in controls. Overall, B types had more VEGF mRNA than A types. VEGF protein levels corresponding to a 23 kDa band were highest in low aldosterone animals in A and C type placentomes. VEGF protein levels corresponding to a 47 kDa band were higher in C type placentomes than A types; protein levels were also higher overall in low cortisol animals compared to controls. Fetoplacental eNOS protein levels were lower in the adrenalectomized groups than in controls. In conclusion, our results indicate that increases in cotyledonary VEGF(164) protein were associated with fetal tissue overgrowth in the placenta when the pregnancy-induced increase in adrenal steroids was prevented in the ewe. However, cotyledonary eNOS protein was suppressed with reduced maternal adrenal steroids, which is consistent with the reduced placental perfusion previously observed in this model. PMID- 17113147 TI - Placental ABCA1 expression is reduced in primary antiphospholipid syndrome compared to pre-eclampsia and controls. AB - The ATP binding cassette transporter A1 (ABCA1) mediates cellular cholesterol and phospholipid efflux, and is implicated in phosphatidylserine translocation and apoptosis. Loss of functional ABCA1 in null mice results in severe placental malformation. This study aimed to establish the placental localisation of ABCA1 and to investigate whether ABCA1 expression is altered in placentas from pregnancies complicated by pre-eclampsia and antiphospholipid syndrome. ABCA1 mRNA and protein localisation studies were carried out using in situ hybridization and immunohistochemistry. Comparisons of gene expression were performed using real-time PCR and immunoblotting. ABCA1 mRNA and protein was localised to the apical syncytium of placental villi and endothelia of fetal blood vessels within the villi. ABCA1 mRNA expression was reduced in placentas from women with APS when compared to controls (p<0.001), and this was paralleled by reductions in ABCA1 protein expression. There were no differences in ABCA1 expression between placentas from pre-eclamptic pregnancies and controls. The localisation of ABCA1 in human placenta is consistent with a role in cholesterol and phospholipid transport. The decrease in ABCA1 protein in APS may reflect reduced cholesterol transport to the fetus affecting the formation of cell membranes and decreasing the level of substrate available for steroidogenesis. PMID- 17113148 TI - Placental diversity in malagasy tenrecs: placentation in shrew tenrecs (Microgale spp.), the mole-like rice tenrec (Oryzorictes hova) and the web-footed tenrec (Limnogale mergulus). AB - Placentation in tenrecs of the subfamily Oryzorictinae, family Tenrecidae, has not been described previously. The structure of the placenta of this group and especially of the genus Microgale was investigated to determine its similarity or dissimilarity to previously described placentas of the tenrec subfamilies Potamogalinae and Tenrecinae. Fifteen specimens of the genus Microgale ranging from an early yolk sac stage to near term were available for study. Placentation in Microgale was found to be different from other tenrecids in that there is an early simple lateral rather than central haemophagous region. In addition, a more villous portion of the placental disk forms before the formation of a more compact labyrinth. Although the definitive placenta is cellular haemomonochorial, it lacks the spongy zone found in the Tenrecinae. Neither does it resemble the endotheliochorial condition found in the Potamogalinae. Of the two genera of the subfamily Oryzorictinae represented by single specimens, the placenta of Limnogale resembled that of the Microgale but Oryzorictes had several differences including a lobulated placental disk. It is concluded that there is more variation in placentation both within the subfamily Oryzorictinae and within the family Tenrecidae than would ordinarily be expected. PMID- 17113149 TI - Abnormal stromal cells in myelodysplastic syndromes: genomics presents further evidence. PMID- 17113150 TI - In silico prediction of FVIII epitopes recognised by natural autoantibodies in polyvalent immunoglobulin concentrates. AB - Inhibitory antibodies directed against blood coagulation factor VIII (FVIII) impair FVIII replacement therapy, constituting a serious complication in haemophilic and autoimmune patients. Identifying B-cell FVIII epitopes and mapping them on the molecule remain important challenges. Using a combination of different algorithms, more than 30 hypothetical linear epitopes were predicted on the FVIII molecule surface. We selected several major predicted sequences, spanning all FVIII domains, for specific antibody induction in rabbits. All peptides tested successfully induced production of specific anti-FVIII rabbit antibodies, supporting the relevance of our approach. To investigate the presence of FVIII-reactive antibodies in the healthy donor population, a pooled fraction rich in all IgG subclasses was purified on peptide-Sepharose columns. Substantial amounts of Ig, specific for each FVIII peptide, were purified with yields ranging from 8 to 223 ng/mg immunoglobulins. Our results confirm the diversity of FVIII epitopes recognised by natural human anti-FVIII autoantibodies. All IgG subclasses were found in the affinity-isolated anti-peptide material, with overrepresentation of IgG2 and IgG4. Evidence was also found for new FVIII epitopes. Five human anti-peptide preparations displayed FVIII-neutralising activity, ranging from 1.3 to 5.3 BU/mg. Although the presence of naturally occurring anti-FVIII antibodies in healthy donors has been previously described, our methodology has allowed, for the first time, a fine mapping of several inhibitory and non-inhibitory epitopes. Our observations support the hypothesis that FVIII inhibitors in haemophilia A and autoimmune disease may originate from the proliferation of natural FVIII-specific B-cell clones. PMID- 17113151 TI - Prevention of stroke and dementia by statin therapy: experimental and clinical evidence of their pleiotropic effects. AB - Stroke and dementia are major causes of disability in most countries. Epidemiological studies have demonstrated that statins (3-hydroxy-3 methylglutaryl coenzyme A reductase inhibitors) are likely to reduce the risk for developing these formidable disorders. The favorable outcomes in statin users may be attributable to not only cholesterol-dependent actions, but also various cholesterol-independent actions called "pleiotropic effects." Several clinical trials have suggested that statins decrease the incidence of stroke, especially ischemic stroke. Statins improve endothelial function, inhibit platelet activation, reduce blood coagulability, and suppress inflammatory reactions, all of which may contribute to the beneficial effects of the therapy. Statins also reduce the risk of vasospasm caused by subarachnoid hemorrhage (SAH). In addition, statins might inhibit the development and progression of Alzheimer's disease (AD), the dominant type of dementia in most industrialized countries, upstream of the amyloid cascade. In vitro studies have shown that statins modulate the metabolism of the beta-amyloid precursor protein (APP) and reduce the extracellular level of its proteolytic product, amyloid-beta (Abeta). The aggregated Abeta is cytotoxic, leading to formation of neurofibrillary tangles and neuronal loss in the brain. Inflammatory processes are active in AD and may contribute significantly to AD pathology. We review the experimental background regarding the pleiotropic effects of statins and summarize clinical trials that examined the preventative effects of statin therapy on stroke and dementia. We include current trials in which statin therapy is initiated within 24 hr of onset of acute ischemic stroke. PMID- 17113152 TI - Determination of drugs in biological fluids by direct injection of samples for liquid-chromatographic analysis. AB - The analysis of drugs in various biological fluids is an important criterion for the determination of the physiological performance of a drug. After sampling of the biological fluid, the next step in the analytical process is sample preparation. The complexity of biological fluids adds to the challenge of direct determination of the drug by chromatographic analysis, therefore demanding a sample preparation step that is often time-consuming, tedious, and frequently overlooked. However, direct on-line injection methods offer the advantage of reducing sample preparation steps and enabling effective pre-concentration and clean-up of biological fluids. These procedures can be automated and therefore reduce the requirements for handling potentially infectious biomaterial, improve reproducibility, and minimize sample manipulations and potential contamination. The objective of this review is to present an overview of the existing literature with emphasis on advances in automated sample preparation methods for liquid chromatographic methods. More specifically, this review concentrates on the use of direct injection techniques, such as restricted-access materials, turbulent flow chromatography and other automated on-line solid-phase extraction (SPE) procedures. It also includes short overviews of emerging automated extraction phase technologies, such as molecularly imprinted polymers, in-tube solid-phase micro-extraction, and micro-extraction in a packed syringe for a more selective extraction of analytes from complex samples, providing further improvements in the analysis of biological materials. Lastly, the outlook for these methods and potential new applications for these technologies are briefly discussed. PMID- 17113153 TI - Automated sample treatment by flow techniques prior to liquid-phase separations. AB - Sample preparation (SP) is an integral and important part of an analytical process. Lately, SP has been the topic of increased interest in research and development of novel advanced technologies. Major needs in this aspect are miniaturization, automation, and enrichment. Among other methodologies, flow techniques can be conveniently and effectively coupled to liquid-phase separation technologies for on-line sample preparation. This paper reviews the current trends in on-line automated sample preparation by flow-through techniques prior to liquid-phase separations. Strategies and interfaces developed to couple flow techniques with liquid chromatography, capillary electrophoresis, and mass spectrometry are described. Advantages and limitations of the coupling are discussed. The paper also highlights certain selected applications of these coupled systems. PMID- 17113154 TI - Digitised spirography as an evaluation tool for intention tremor in multiple sclerosis. AB - This study investigated validity and reliability of digitised circle and square spiral drawing for quantifying intention tremor severity and related disability in patients with multiple sclerosis (MS). The tremor amplitude was measured as the standard deviation of the drawing velocity of the arm in the radial and tangential direction for circle spiral drawing, and in the horizontal and vertical direction for square spiral drawing. Results were compared with those of MS patients without tremor and healthy controls, and correlated with clinical assessments of tremor severity and arm functionality including Fahn's tremor rating scale, Test d'Evaluation des Membres superieurs des Personnes Agees (TEMPA) and the nine-hole-peg test to examine validity. Comparison of patient's performance between four repeated trials examined short-term test-retest reliability. All digitised spirography variables discriminated between the MS tremor and both MS-no-tremor and healthy control groups. Validity was also shown by high spearman correlation coefficients between spirography variables and clinical ratings. Tremor appeared to be most profound in the radial and vertical direction during circle and square spiral drawing, respectively. The consistency and high correlations between four repeated executions indicated short-term test retest reliability. We conclude that the digitised spirography provide a useful instrumentation for quantifying MS intention tremor. PMID- 17113155 TI - A family study of co-morbidity between generalized social phobia and generalized anxiety disorder in a non-clinic sample. AB - BACKGROUND: High rates of co-morbidity between Generalized Social Phobia (GSP) and Generalized Anxiety Disorder (GAD) have been documented. The reason for this is unclear. Family studies are one means of clarifying the nature of co-morbidity between two disorders. METHODS: Six models of co-morbidity between GSP and GAD were investigated in a family aggregation study of 403 first-degree relatives of non-clinical probands: 37 with GSP, 22 with GAD, 15 with co-morbid GSP/GAD, and 41 controls with no history of GSP or GAD. Psychiatric data were collected for probands and relatives. Mixed methods (direct and family history interviews) were utilised. RESULTS: Primary contrasts (against controls) found an increased rate of pure GSP in the relatives of both GSP probands and co-morbid GSP/GAD probands, and found relatives of co-morbid GSP/GAD probands to have an increased rate of both pure GAD and co-morbid GSP/GAD. Secondary contrasts found (i) increased GSP in the relatives of GSP only probands compared to the relatives of GAD only probands; and (ii) increased GAD in the relatives of co-morbid GSP/GAD probands compared to the relatives of GSP only probands. LIMITATIONS: The study did not directly interview all relatives, although the reliability of family history data was assessed. The study was based on an all-female proband sample. The implications of both these limitations are discussed. CONCLUSIONS: The results were most consistent with a co-morbidity model indicating independent familial transmission of GSP and GAD. This has clinical implications for the treatment of patients with both disorders. PMID- 17113156 TI - Evaluating the validity of blood-based membrane potential changes for the identification of bipolar disorder I. AB - OBJECTIVE: The objective of this study was to develop a diagnostic blood test for bipolar disorder I using membrane potentials as biological markers. METHODS: We measured the fluorescence intensity of a dye sensitive to membrane potential in whole blood samples from bipolar I, unipolar, schizophrenic patients, and psychiatrically normal controls. Patients were diagnosed through structured clinical interviews according to DSM-IV. Both the t-test and logistic regression analysis were used to analyze the data. RESULTS: The membrane potential as indicated by the fluorescence intensity of the membrane potential dye in blood cells drawn from patients with bipolar disorder I was significantly different from the blood cells drawn from unipolar and schizophrenic patients, and from psychiatrically normal controls (P<0.001). The specificity and sensitivity were determined to be 0.88 and 0.78 respectively which compared well with the state of the art diagnostic techniques for other diseases. Logistic regression analysis revealed that the membrane potential was a reliable predictor which could be used as a diagnostic marker for bipolar I. CONCLUSIONS: These results indicate that the membrane potential of blood cells can be used as a diagnostic marker to augment the DSM-IV diagnosis of bipolar disorder I. Expanded clinical trials are needed to establish this technique for general use. PMID- 17113157 TI - Frontal functions in depressed and nondepressed Parkinson's disease patients: impact of severity stages. AB - Severity of Parkinson's disease (PD) and frontal impairment are positively correlated. Testing frontal functions in depressed/nondepressed PD patients with different severity stages may reveal whether depression leads to this impairment. We aimed to relate severity of PD to frontal functional impairment and to test if negative stimuli/depressive symptoms interfered with frontal tasks. The Stroop test and the Emotional Stroop test were performed by 46 PD patients, 18 of whom were depressed. The Hoehn and Yahr scale assessed severity of the disease. We calculated the difference in seconds for each Stroop card and the interference index (C/D) between depressed and nondepressed patients sharing the same severity of disease. The differences among the groups (depressed and nondepressed) according to the severity of the disease (mild and moderate) were compared using the Mann-Whitney test. The depressed patients had a poorer performance on the test than the nondepressed PD patients, although the difference was not statistically significant. In conclusion, there is a clinically relevant but not statistically significant difference on the performance of frontal tasks between depressed and nondepressed PD patients. Neither depression nor the severity of the disease were determinant to the poorer performance on the Stroop and the Emotional Stroop tests. PMID- 17113158 TI - Plasma homovanillic acid correlates inversely with history of learning problems in healthy volunteer and personality disordered subjects. AB - Central dopaminergic activity is critical to the functioning of both motor and cognitive systems. Based on the therapeutic action of dopaminergic agents in treating attention deficit hyperactivity disorder (ADHD), ADHD symptoms may be related to a reduction in central dopaminergic activity. We tested the hypothesis that dopaminergic activity, as reflected by plasma homovanillic acid (pHVA), may be related to dimensional aspects of ADHD in adults. Subjects were 30 healthy volunteer and 39 personality disordered subjects, in whom morning basal pHVA concentration and a dimensional measure of childhood ADHD symptoms (Wender Utah Rating Scale: WURS) were obtained. A significant inverse correlation was found between WURS Total score and pHVA concentration in the total sample. Among WURS factor scores, a significant inverse relationship was noted between pHVA and history of "childhood learning problems". Consistent with the dopaminergic dysfunction hypothesis of ADHD and of cognitive function, pHVA concentrations were correlated with childhood history of ADHD symptoms in general and with history of "learning problems" in non-ADHD psychiatric patients and controls. Replication is needed in treated and untreated ADHD samples to confirm these initial results. PMID- 17113159 TI - Psychological distress among American Red Cross disaster workers responding to the terrorist attacks of September 11, 2001. AB - This study investigated American Red Cross disaster workers' symptoms of distress and posttraumatic stress resulting from exposure to disaster stimuli during their response to the September 11, 2001 terrorist attacks. A sample of 3055 Red Cross disaster workers was surveyed 1 year after the terrorist attacks regarding demographic characteristics, function during the response, and exposure to disaster stimuli. Participants were grouped by function and self-reported exposure, with the hypothesis that workers in Direct Services and/or those reporting to be directly exposed to disaster stimuli would experience greater levels of posttraumatic stress symptoms and distress than workers in indirect services or reporting no exposure. Findings revealed that while there were significant differences between both Function and Exposure groups on dependent measures, the multivariate eta2 was very small for both and did not meet medium effect size criteria. The results indicated that workers directly exposed to disaster stimuli reported no more distress than those who were not directly exposed. PMID- 17113160 TI - Gene-environment interactions in multiple sclerosis: innate and adaptive immune responses to human endogenous retrovirus and herpesvirus antigens and the lectin complement activation pathway. AB - Aspects of gene-environment interactions in multiple sclerosis (MS) were analysed in serum samples from 46 MS families (25 sporadic MS cases and 42 familial MS cases): antibodies to the MS-associated human endogenous retrovirus HERV-H, and levels of three components in the innate pathogen-associated molecular pattern recognition: mannan-binding lectin (MBL), and MASP-2 and MASP-3. For representative MS families, we also determined herpesvirus serology for HSV-1, VZV, and EBV; and tissue typed for HLA-B, and HLA DR and DQ. In MS, a significant correlation between elevated immune reactivity to HERV-H Env and disease activity was demonstrated, as were indications of a protective effect of high MBL and MASP 3 levels. The HLA alleles B*07, DRB*02, and DQB1*06 were commonly present together in the MS families, both in MS patients, and in unaffected family members. Our results support that HERV-H and the antiviral immune response may play a role in MS development, and also underline the tenuous nature of specific genetic contributions to this complex disease. PMID- 17113161 TI - Anti-ganglioside complex antibodies associated with severe disability in GBS. AB - Ganglioside complexes (GSCs) are known as target antigens in Guillain-Barre syndrome (GBS). To elucidate the clinical importance of the anti-GSC antibodies in GBS, we investigated serum antibodies to GSCs containing two of the gangliosides, GM1, GD1a, GD1b and GT1b, and analyzed clinical features of anti GSC-positive GBS patients. Thirty-nine (17%) of 234 GBS patients had IgG anti-GSC antibodies. Anti-GSC-positive GBS had antecedent gastrointestinal infection and lower cranial nerve deficits more frequently than control GBS. The presence of antibody specificity to GD1a/GD1b and/or GD1b/GT1b was significantly associated with severe disability and a requirement for mechanical ventilation. PMID- 17113162 TI - Vestibular symptoms in children with enlarged vestibular aqueduct anomaly. AB - OBJECTIVE: The objective of this study is to describe the vestibular symptoms in pediatric patients with enlarged vestibular aqueduct (EVA) anomaly. METHODS: Retrospective chart review of pediatric and adult patients with EVA anomaly who were treated at the University of Utah Hospital or Primary Children's Medical Center, between 1995 and 2005. Radiographs were reviewed to confirm the diagnosis. Comparisons were made between adult and pediatric patients. RESULTS: Thirty-two patients were included in the study, 17 females and 15 males. Twenty one patients were under the age of 18 and 11 patients were age 18 or older. On initial audiometric evaluation at a tertiary hospital, the pure tone average in the right ear was 75.0 dB and the pure tone average in the left ear was 80.4 dB. The incidence of vestibular symptoms in adult patients was 45.5% and in pediatric patients was 48.0%. Fourteen patients underwent cochlear implantation. Four patients (28.6%) who previously denied vestibular symptoms experienced post operative vertigo after cochlear implantation. CONCLUSIONS: About half of the patients with EVA in our series experienced vestibular symptoms. Pediatric patients in our series experienced vertigo and vestibular symptoms with equal frequency when compared to adult patients. Some patients with EVA undergoing cochlear implantation experienced vestibular symptoms in the post-operative period. PMID- 17113163 TI - ZENK labeling within social behavior brain regions reveals breeding context dependent patterns of neural activity associated with song in male European starlings (Sturnus vulgaris). AB - In songbirds, song learning and production are regulated by the song control system. How the rest of the brain interacts with song nuclei to ensure that song is produced in an appropriate context is not yet clear. In male European starlings (Sturnus vulgaris), breeding context song is sexually motivated, whereas, non-breeding context song is more broadly socially motivated. Brain regions involved in regulating social behavior might differentially regulate starling song depending upon the context in which it is produced. Here, we compared the number of ZENK-labeled cells in song and social behavior nuclei in starlings singing in either a breeding or a non-breeding context. Numbers of ZENK labeled cells in HVC related positively to song produced in both contexts. Interestingly, numbers of ZENK-labeled cells in one subdivision of the lateral septum (LS) related negatively to breeding context song but positively to non breeding context song. In a subdivision of the medial bed nucleus of the stria terminalis (BSTm) ZENK labeling only related positively to non-breeding context song, whereas, in the ventromedial nucleus of the hypothalamus (VMH) ZENK labeling showed a tighter positive relationship with breeding context song. Together, these findings indicate that social behavior brain regions outside of the song control system regulate singing behavior differently depending upon whether song is sexually or more broadly socially motivated. Breeding context dependent regulation of song by LS, BSTm, and VMH suggests that these nuclei may be central to adjusting song production so that it occurs in response to appropriate social and environmental stimuli. PMID- 17113164 TI - Effects of paradoxical sleep deprivation on blood parameters associated with cardiovascular risk in intact and ovariectomized rats compared with male rats. AB - The purpose of this study was to investigate the effects of paradoxical sleep deprivation (PSD) on circulating lipoproteins (total cholesterol, HDL, LDL and triglycerides) in males as well as in intact and ovariectomized (OVX) female rats. The intact female rat group was sub-distributed according to the phase of the estrous cycle (proestrus, estrus and diestrus) allowing for comparison of the lipid profile with males and OVX rats. The results indicate that PSD significantly reduced cholesterol in intact females compared to OVX and male rats; it reduced triglycerides in all groups except in diestrus rats and increased HDL levels in male rats compared with the respective controls. PSD also increased LDL levels in male and OVX rats when compared to intact females. Examinations of cholesterol fractions revealed significant increases in HDL in control-OVX animals when compared to the other groups, whereas HDL was significantly increased after PSD in male rats. Such results suggest that the cardiovascular response in intact, OVX females and male rats is differentially regulated especially when such are submitted to PSD. Similarities in blood parameters observed between OVX and male rats are likely due to the suppression of ovarian hormone release after ovariectomy. PMID- 17113165 TI - Dietary phenolic antioxidants, caffeic acid and Trolox, protect rainbow trout gill cells from nitric oxide-induced apoptosis. AB - Caffeic acid (CA) and Trolox are phenolic acids that have beneficial antioxidant effect, but the underlying mechanisms involved are not fully understood. The extent to which CA and Trolox protect against sodium nitroprusside (SNP)-induced oxidative cell injury was investigated in cultured rainbow trout gill cells. The cells exposed to SNP for 24 h displayed a dose-dependent leakage of lactate dehydrogenase (LDH) and decreased cell viability as indicated by the MTT assay (mitochondrial dehydrogenase activity). Both effects were prevented by treatment with 50 microM CA or Trolox. CA or Trolox, protected against SNP-induced caspase 3 activation and DNA fragmentation, indicating a reduction of apoptosis. Thus, the results indicate that SNP induced cell death is caspase-3 related apoptosis and the treatment with CA inhibited the apoptotic pathway. In addition, we studied the effect of CA and Trolox on expression of zinc-responsive antioxidant genes such as metallothioneins (MT), glutathione-S-transferase (GST Class pi) and glucose-6-phosphate dehydrogenase (G6PD) in cultured gill cells. CA, 100 microM, increased accumulation of mRNA for MTA, MTB, GST and G6PD in cells. Thus, in addition to its ability to sequester free radicals, CA may protect against oxidative stress through expression of zinc-induced antioxidant proteins. Because of these properties we suggest that CA could be a beneficial additive to fish feeds in aquaculture. PMID- 17113166 TI - Effects of binary mixtures of xenoestrogens on gonadal development and reproduction in zebrafish. AB - Previous studies exposing fish to xenoestrogens have demonstrated vitellogenin (VTG) induction, delayed gametogenesis, altered sex ratio, and decreased reproductive performance, with a majority of those studies focusing on exposure to single chemicals. The objective of this study was to determine the effects of binary mixtures of a weak estrogen receptor agonist, nonylphenol (NP) and a potent estrogen receptor agonist, 17alpha-ethinylestradiol (EE) on sex ratios, gametogenesis, VTG induction, heat shock protein 70 (HSP70) expression and reproductive capacity in zebrafish (Danio rerio). Fish were exposed from 2 to 60 days post-hatch (dph) to nominal concentrations of 10 or 100 microg/l NP (NP10 or NP100, respectively), 1 or 10 ng/l EE (EE1 or EE10, respectively), 1 ng/l EE+10 or 100 microg/l NP (EE1+NP10 or EE1+NP100, respectively), 10 ng/l EE+10 or 100 microg/l NP (EE10+NP10 or EE10+NP100, respectively) or solvent control (0.01% acetone, v/v) in a static-renewal system with replacement every 48 h. At 60 dph, fish from each treatment were euthanized for histological examination of gonads, and whole body VTG and HSP70 levels. Remaining fish were reared in clean water until adulthood (240 dph) for breeding studies. In all EE10 exposure groups (EE10, EE10+NP10 and EE10+NP100), increasing NP concentration acted antagonistically to the action of EE in terms of VTG induction at 60 dph. Similarly, non-additivity was observed with egg production, where EE1+NP100 exposure resulted in significantly more eggs produced per breeding trial than EE1 alone. Histological staging of oogenesis revealed suppressed gametogenesis in an additive fashion in females at 60 dph. There were no differences among treatment groups in whole body HSP70 expression in 60 dph fish or in gonadal HSP70 expression in adult fish. Although there was no statistical evidence of non additivity, breeding trials in adults revealed significant reductions in egg viability, egg hatchability and/or F1 swim-up success, suggesting that developmental exposures to xenoestrogens may cause irreversible effects on egg quality and progeny even after periods of depuration. In conclusion, these results suggest that environmentally relevant mixtures of NP and EE can produce additive or non-additive effects that depend on the particular response being determined and the respective exposure concentrations of each chemical. PMID- 17113167 TI - The amyloid precursor protein potentiates CHOP induction and cell death in response to ER Ca2+ depletion. AB - Here we investigated the role of the amyloid precursor protein (APP) in regulation of Ca(2+) store depletion-induced neural cell death. Ca(2+) store depletion from the endoplasmic reticulum (ER) was induced by the SERCA (Sarco/Endoplasmic Reticulum Calcium ATPase) inhibitor thapsigargin which led to a rapid induction of the unfolded protein response (UPR) and a delayed activation of executioner caspases in the cultures. Overexpression of APP potently enhanced cytosolic Ca(2+) levels and cell death after ER Ca(2+) store depletion in comparison to vector-transfected controls. GeneChip and RT-PCR analysis revealed that the expression of classical UPR chaperone genes was not altered by overexpression of APP. Interestingly, the induction of the ER stress-responsive pro-apoptotic transcription factor CHOP was significantly upregulated in APP overexpressing cells in comparison to vector-transfected controls. Chelation of intracellular Ca(2+) with BAPTA-AM revealed that enhanced CHOP expression after store depletion occurred in a Ca(2+)-dependent manner in APP-overexpressing cells. Prevention of CHOP induction by BAPTA-AM and by RNA interference was also able to abrogate the potentiating effect of APP on thapsigargin-induced apoptosis. Application of the store-operated channel (SOC)-inhibitors SK & F96365 and 2-APB downmodulated APP-triggered potentiation of cytosolic Ca(2+) levels and apoptosis after treatment with thapsigargin. Our data demonstrate that APP significantly modulates Ca(2+) store depletion-induced cell death in a SOC- and CHOP-dependent manner, but independent of the UPR. PMID- 17113168 TI - Heart rate variability in beta-thalassemic mice. PMID- 17113169 TI - Prognostic significance of exercise stress echocardiography in 3329 outpatients (5-year longitudinal study). AB - BACKGROUND: Appraisal of the risk to which outpatients with chest pain are exposed is a major clinical problem. Up to now, there have been no reports on the prognostic significance of exercise stress echocardiography in this patient cohort. PATIENTS AND METHODS: In order to investigate the prognostic significance of exercise stress echocardiography (SE) in outpatients only, 3329 patients were monitored during a long-term follow-up regarding the occurrence of hard events (cardiac death, myocardial infarction, revascularization). The patients came to the cardiology practice complaining of chest pain. RESULTS: The sensitivity/specificity of SE for hard events was 81.1/92.8 in the first year, that of exercise ECG, 27.4/87.0. During the observation period (5.1+/-1.1 years (median 5.2, 3-7 years)), a total of 446 (13.4%) hard events occurred. In patients with positive SE findings, 262 (61.9%) hard events occurred, in patients with negative SE findings, hard events were rarer (184, 6.3%, p<0.001). In the multivariate analysis, the positive SE finding was the most unambiguous, significant independent predictor of hard events (HR 6.6, CI 5.21-8.25, p<0.001). CONCLUSIONS: In outpatients with chest pains, exercise stress echocardiography is of major prognostic significance (independent of other parameters) and its prognostic reliability is clearly superior to that of the exercise ECG. SE should always be performed in cases with symptoms requiring clarification. PMID- 17113170 TI - Candesartan in the prevention of relapsing atrial fibrillation. AB - BACKGROUND: Several studies have indicated that treatment with angiotensin converting enzyme (ACE) inhibitors and angiotensin II type 1 receptor blockers (ARBs) may reduce the incidence of atrial fibrillation (AF) in hypertensive patients and patients with left ventricular dysfunction. However, there is limited data on the effect of ACE-inhibitors and ARBs in patients undergoing electrical cardioversion for persistent AF. We hypothesized that treatment with the ARB candesartan, without adjunct antiarrhythmic therapy, would reduce the recurrence rate of AF after successful cardioversion. METHODS: In a double blind, placebo-controlled study, 171 patients with persistent AF were randomized to receive candesartan 8 mg once daily (n=86) or placebo (n=85) for 3-6 weeks before and candesartan 16 mg once daily or placebo for 6 months after electrical cardioversion. Primary endpoint was recurrence of AF. RESULTS: A total of 68 patients in the candesartan group and 69 patients in the placebo group were successfully cardioverted. Forty-eight patients (71%) in the candesartan group and 45 (65%) in the placebo group had a recurrence of AF during 6 months follow up. Median time to recurrence was 8 and 9 days in the candesartan and placebo groups, respectively. The differences between the groups were not statistically significant. CONCLUSION: Treatment with the ARB candesartan for 3-6 weeks before and 6 months after electrical cardioversion had no effect on the recurrence rate of AF. PMID- 17113171 TI - Incomplete myocardial rupture following inferior myocardial infarction: a case report. AB - In an era of early and invasive therapeutic approaches, myocardial rupture has become an uncommon complication of myocardial infarction. While septal wall rupture most often leads to devastating haemodynamic consequences, free wall rupture is usually fatal. We report a case of a 48-year-old man in whom an incomplete myocardial rupture located in the inferior part of the interventricular septum was promptly detected during the acute phase of an inferior myocardial infarction treated by early percutaneous coronary angioplasty. A conservative rather than a surgical approach was decided with a favourable short-term outcome. PMID- 17113172 TI - Should Clinical Investigators invEnt acroNyms that are CoercivE?--The SCIENCE in medicine study. PMID- 17113173 TI - HIV prevalence and poverty in Africa: micro- and macro-econometric evidences applied to Burkina Faso. AB - Based on the data of the Demographic and Health Survey, and of the Household Priority Survey, carried out in 2003, the present study, examining the factors of HIV prevalence in Burkina Faso, provides two conclusions. Firstly, the fight against poverty is not necessarily a means of reducing simultaneously and drastically HIV/AIDS prevalence, an assertion based on several elements of empirical analysis. First of all, the micro-econometric estimates of the probit models suggest a positive relationship between HIV prevalence in adult women and men, and living standards of individuals. Then, the macro-econometric approach reveals the existence of a positive (negative) relationship between, on the one hand, the level of regional HIV prevalence, and, on the other hand, the average monetary provincial standard of living (poverty) of households. At the same time, the relationship between HIV prevalence and poverty, apprehended at the regional level, is not linear. Secondly, and correlatively, the relationship between HIV prevalence and poverty is called into question. First of all, some structural factors may contribute to a distortion of the relationship between resources of households and the prevalence of HIV/AIDS. This may be due, on the one hand, to the persistence of cognitive and behavioural factors inherent in a traditional society, and in particular, to the fact that the social construction of female attributes and roles confers to men a statute of "decision-makers" with regard to sexual intercourse, while the persistence of secular beliefs contributes to minimizing the perception of HIV/AIDS in terms of risk, independently of standards of living. In addition, the enclavement of Burkina Faso required development of road and railway traffic with neighbouring countries, in particular Cote d'Ivoire. Therefore, it may be that the structural conditions of the process of development of Burkina Faso, concomitant with significant flows of the exchange of goods, services and labour with a country where the prevalence of the HIV is particularly high, constitute an element of an explanation of the positive relationship between the resources of households and HIV seroprevalence. Also, factors related to the economic situation probably contributed to reinforcing the opposite relationship between HIV seroprevalence and poverty, the macro-econometric analysis highlighting a direct relationship between the massive return of migrants of Cote d'Ivoire and the level of HIV prevalence in Burkina Faso. PMID- 17113174 TI - Web-based education in bioprocess engineering. AB - The combination of web technology, knowledge of bioprocess engineering, and theories on learning and instruction might yield innovative learning material for bioprocess engineering. In this article, an overview of the characteristics of web-based learning material is given, as well as guidelines for the design of learning material from theories of learning and instruction and from the bioprocess engineering domain. A diverse body of learning material is presented, which illustrates the application of these guidelines; this material has been developed during the past six years for different courses, mostly at undergraduate level, and it illustrates how web-based learning material can enable various different approaches to learning objectives that might improve overall learning. Such learning material has been used for several years in education, it has been evaluated with positive results, and is now part of the regular learning material for bioprocess engineering at Wageningen University. PMID- 17113175 TI - Linkage disequilibrium analysis of the CHRNA7 gene and its partially duplicated region in schizophrenia. AB - Several previous studies have reported a significant linkage between markers in the alpha 7 nicotinic cholinergic receptor subunit (CHRNA7) gene and either schizophrenia or the P50 sensory gating deficit, a schizophrenia endophenotype. However, CHRFAM7A, a partially duplicated gene 1.6Mb upstream of the CHRNA7 gene, has complicated further genetic analysis. We genotyped 14 polymorphic markers throughout the full-length CHRNA7 gene and the duplicated region in 188 unrelated Han Chinese patients with schizophrenia and 188 controls. The duplicated regions were assessed by genotyping up- and down-stream polymorphic markers in the vicinity of each region and analyzing the linkage disequilibrium (LD) between each pair of markers. No evidence of risk variants for schizophrenia in either the CHRNA7 gene or the partially duplicated region was found in the LD analysis. A significant deviation from the Hardy-Weinberg equilibrium (HWE) was found only in the genotypic distribution of SNP9 (IVS4-1912) in patients (p=0.00829), but not in controls. In conclusion, our LD analysis did not reveal any association between schizophrenia in our Han Chinese population and the CHRNA7 gene or its partially duplicated region. However, we could not exclude the possibility of a weak genetic effect due to the small sample size. Analyses of larger samples and higher-density markers, particularly around SNP9 (IVS4-1912), are still needed. PMID- 17113176 TI - Gene delivery by combination of novel liposomal bubbles with perfluoropropane and ultrasound. AB - Microbubbles and ultrasound have recently been investigated with a view to improving the transfection efficiency of non-viral gene delivery systems. However, microbubbles are unstable and their targeting ability is insufficient for clinical use. To circumvent these problems, we developed novel polyethyleneglycol (PEG) modified liposomes (Bubble liposomes) containing perfluoropropane, which is an ultrasound imaging gas. Here, we used ultrasound to induce cavitation in Bubble liposomes and then investigated their ability to deliver genes in vitro and in vivo. Bubble liposomes could deliver plasmid DNA to many cell types without cytotoxicity. Additionally, in vivo gene delivery, Bubble liposomes were more effective delivery into femoral artery than lipofection method. Thus, Bubble liposomes might be efficient and novel non-viral tools for gene delivery. PMID- 17113177 TI - Impact of intraperitoneal, sustained delivery of paclitaxel on the expression of P-glycoprotein in ovarian tumors. AB - Recently, we developed a novel implantable drug delivery system which can provide sustained intraperitoneal (i.p.) delivery of paclitaxel (PTX). As the impact of local sustained delivery on the development of multidrug resistance (MDR) is unknown, the objective of this study was to determine the impact of this drug delivery system on the in vivo expression of MDR1/P-glycoprotein (PGP) in a human ovarian xenograft tumor model. As compared to controls, intermittent i.p. dosing with PTX formulated in Cremophor EL (PTX(CrEL)) induced a two-fold increase in mRNA levels of MDR1 after a 14-day dosing period. On the other hand, sustained i.p. delivery of PTX with the implant system (PTX(film)) did not significantly affect MDR1 expression. Immunodetection of PGP in isolated xenografts supported the mRNA data. Histological analysis by H&E staining demonstrated a dose dependent increase in tumor necrosis in the PTX(film) treated animals. Further, in vitro studies in human ovarian carcinoma cells also demonstrated a significant induction in the efflux activity of PGP with intermittent dosing schedules to PTX(CrEL) whereas this was not seen in cells dosed with PTX(film). Our findings suggest that sustained i.p. administration with PTX(film) attenuates development of MDR, suggesting that sustained, localized delivery of chemotherapeutic agents may improve current treatment strategies for ovarian cancer. PMID- 17113178 TI - Hybrid polymer nanocapsules enhance in vitro delivery of azidothymidine triphosphate to macrophages. AB - One of the main limitations in the use of nucleoside reverse transcriptase inhibitors (NRTIs) such as azidothymidine (AZT) lies in their poor intracellular activation by cellular kinases into their active tri-phosphorylated form. Thus, the direct administration of triphosphate NRTIs like azidothymidine-triphosphate (AZT-TP), has been considered for bypassing this metabolic bottleneck, but these molecules do not diffuse intracellularly, due to their too hydrophilic character. Therefore, poly(iso-butylcyanoacrylate) (PIBCA) aqueous-cored nanocapsules have been tested as carriers to overcome the cellular delivery of AZT-TP. However, encapsulation of AZT-TP remained challenging because this molecule, due to its relatively low molecular weight, rapidly leaked out of the nanocapsules. In this study, we show that association of AZT-TP to a cationic polymer such as poly(ethyleneimine) (PEI) allowed to reach high entrapment efficiency of AZT-TP in PIBCA nanocapsules (up to 90%) as well as gradual in vitro release. The resulting hybrid PIBCA/PEI nanocapsules efficiently delivered AZT-TP in vitro to macrophages: the cellular uptake was increased by 30-fold compared to the free molecule, reaching relevant cellular concentrations for therapeutic purposes. PMID- 17113179 TI - A prospective study comparing insulin and glibenclamide in gestational diabetes mellitus in Asian Indian women. PMID- 17113180 TI - Public responses to precautionary information from the Department of Health (UK) about possible health risks from mobile phones. AB - Understanding public perceptions of health information is of increasing importance in the light of the growing imperatives upon regulators to communicate information about risk and uncertainty. Communicating the possible health risks from mobile telecommunications is a domain that allows consideration of both public perceptions of uncertain public health information and public responses to precautionary advice. This research reports the results of a nationally representative survey in the UK (n=1742) that explored public responses to a leaflet issued by the Department of Health (DoH) in 2000 providing information about the possible health risks of mobile phones. The aims of the study were two fold: (a) to assess awareness of the leaflet and the extent to which participants could identify the precautionary advice that the leaflet contained as coming from the Government; and (b) to examine publics' responses to the current Government precautionary advice about mobile phone health risks; was this associated with increased concern or reassurance? The results indicate the importance of policy makers developing a clear understanding of the possible effects of communicating precautionary advice. PMID- 17113181 TI - Integrating digital topology in image-processing libraries. AB - This paper describes a method to integrate digital topology informations in image processing libraries. This additional information allows a library user to write algorithms respecting topological constraints, for example, a seed fill or a skeletonization algorithm. As digital topology is absent from most image processing libraries, such constraints cannot be fulfilled. We describe and give code samples for all the structures necessary for this integration, and show a use case in the form of a homotopic thinning filter inside ITK. The obtained filter can be up to a hundred times as fast as ITK's thinning filter and works for any image dimension. This paper mainly deals of integration within ITK, but can be adapted with only minor modifications to other image-processing libraries. PMID- 17113182 TI - Simple multichannel system for the measurement of the net water flux across biological tissues. AB - This paper describes the development of a simple system for measurement of net water movement through biological membrane barriers. The system is based on the detection of a water meniscus inside a polyethylene tube, which reflects the water movement inside one hemichamber of a modified Ussing chamber containing a membrane barrier. The detection device consists of a commercial computer controlled flat bed scanner and specifically developed software. This system allows one to perform a relatively high number of individual experiments per physical unit. It is a flexible and affordable device, which allows comparatively more information per unit to be obtained than previously described methods. PMID- 17113183 TI - CASANDRA: a prototype implementation of a system of network progressive transmission of medical digital images. AB - In this paper, a prototype for progressive transmission of medical digital 2D images through the network, called CASANDRA, is presented. The prototype consists of the server part and the client part. In the server part, the images are acquired, stored, computed their wavelet transform and the wavelet coefficients stored, then transmitted progressively, when required, via TCP to the client. In the client part, with the inverse wavelet transform, the received wavelet coefficients are used to build successive improved reconstructions of the image. This prototype has been implemented and is being tested in the Radiotherapy Service of the Valencia University Hospital (Valencia, Spain). PMID- 17113184 TI - Targeting lung cancer using an infectivity enhanced CXCR4-CRAd. AB - Conventional treatments are not adequate for the majority of lung cancer patients. Conditionally replicating adenoviruses (CRAds) represent a promising new modality for the treatment of neoplastic diseases, including non-small cell lung cancer. Specifically, following cellular infection, the virus replicates selectively in the infected tumor cells and kills the cells by cytolysis. Next, the progeny virions infect a new population of surrounding target cells, replicate again and eradicate the infected tumor cells while leaving normal cells unaffected. However, to date, there have been two main limitations to successful clinical application of these CRAd agents; i.e. poor infectivity and poor tumor specificity. Here we report the construction of a CRAd agent, CRAd-CXCR4.RGD, in which the adenovirus E1 gene is driven by a tumor-specific CXCR4 promoter and the viral infectivity is enhanced by a capsid modification, RGD4C. This agent CRAd CXCR4.RGD, as expected, improved both of the viral infectivity and tumor specificity as evaluated in an established lung tumor cell line and in primary tumor tissue from multiple patients. As an added benefit, the activity of the CXCR4 promoter was low in human liver as compared to three other promoters regularly used for targeting tumors. In addition, this agent has the potential of targeting multiple other tumor cell types. From these data, the CRAd-CXCR4.RGD appears to be a promising novel CRAd agent for lung cancer targeting with low host toxicity. PMID- 17113185 TI - Safety and feasibility of cardiopulmonary exercise testing in patients with advanced cancer. AB - PURPOSE: The use of cardiopulmonary exercise testing (CPET) is increasing in many areas of disease management, the clinical utility of this procedure in patients with advanced cancer remains to be determined. Accordingly, we conducted a pilot study to comprehensively evaluate the safety and feasibility of CPET in 85 patients diagnosed with inoperable non-small cell lung cancer (NSCLC) or metastatic breast cancer (MBC). METHODS: Using a cross-sectional design, consecutive patients with histologically or cytologically confirmed inoperable NSCLC or MBC were screened for eligibility by their attending oncologist. All consenting patients performed complete pulmonary function tests (NSCLC only) and a symptom-limited CPET on a cycle ergometer with gas exchange analysis. RESULTS: A total of 85 patients (NSCLC, n=46; MBC, n=39) were recruited and tested. Overall, there were a total of three (3.5%) positive exercise tests [NSCLC, n=2 (4.3%); MBC, n=1 (2.6%)]; 68 (80.0%) negative tests [NSCLC, n=33 (71.7%); MBC, n=35 (89.7%)]; and 14 (16.5%) indeterminate tests [NSCLC, n=11 (23.3%); MBC, n=3 (7.7%)]. Two patients experienced an adverse event during exercise testing. Mean VO(2peak) was 17.0mlkg(-1)min(-1) and 16.5mlkg(-1)min(-1) for NSCLC and MBC patients, respectively. The mean percentage of age and sex-predicted VO(2peak) maximum for both groups was 67%. CONCLUSION: A symptom-limited, individualized CPET appears to be a relatively safe and feasible assessment tool to objectively evaluate physical functioning in selected patients with advanced cancer. This study provides important information to future investigations examining the potential role of exercise training in this patient population. PMID- 17113186 TI - Gastrointestinal solitary metastases from squamous cell lung cancer. PMID- 17113187 TI - Clinical and immunopathologic findings during treatment of recalcitrant atopic eczema with efalizumab. AB - Treatment of widespread moderate to severe atopic eczema remains a challenge. The therapeutic efficacy and modifications of the immune response during treatment of atopic eczema with efalizumab are so far unknown. We hereby report the clinical findings and characterize the inflammatory infiltrate during treatment of severe recalcitrant atopic eczema with efalizumab. PMID- 17113188 TI - Oral histoplasmosis after radiation therapy for laryngeal squamous cell carcinoma. AB - Histoplasmosis is a usually asymptomatic deep fungal infection of tropical origin with respiratory entry and possible oral, pharyngeal, or metastatic localization. The condition represents an important imported systemic mycosis with oral involvement. We report the case of a patient who developed an oropharyngeal reactivation of a latent Histoplasma infection after receiving local antitumoral radiation therapy of the neck. H capsulatum was shown to be present in the lesion by both histopathology and staining, and was deduced to be the causative organism of the disease. PMID- 17113189 TI - Dermoscopy features of melanoma incognito: indications for biopsy. AB - BACKGROUND: To avoid missing melanoma, the current practice is to biopsy all suggestive skin lesions. Although most cases of melanoma exhibit clinical clues leading to the correct diagnosis, melanoma can mimic benign lesions. Dermoscopy has been shown to increase the diagnostic accuracy of clinically equivocal lesions, but little is known about its ability to detect melanoma in the context of lesions that appear clinically benign. METHODS: We present 7 difficult-to diagnose melanomas, in which additional clues provided by dermoscopy increased the index of suggestion and led us to perform a biopsy. RESULTS: Our cases highlight the following 7 management rules: 1) Dermoscopy should not be used only for suggestive skin lesions. 2) Biopsy lesions missing clinicodermoscopic correlation. 3) Biopsy lesions with unspecific pigment pattern. 4) Biopsy lesions with spitzoid features. 5) Biopsy lesions with extensive regression features. 6) In patients with multiple nevi, biopsy lesions changing after short-term follow up. 7) Biopsy pink lesions with an atypical vascular pattern. LIMITATIONS: The reported series of cases is small. Dermoscopy has not been rigorously compared with handheld magnification (as with a x7 loupe). CONCLUSIONS: Dermoscopy can increase the index of suggestion to perform biopsy in difficult-to-diagnose melanomas. PMID- 17113190 TI - A randomized, open-label trial of continuous versus interrupted etanercept therapy in the treatment of psoriasis. AB - BACKGROUND: Although etanercept is used as a continuous therapy for moderate to severe plaque psoriasis, intermittent use may be necessary in some instances. OBJECTIVE: In this randomized, open-label study, we evaluated the effectiveness and safety of continuous versus interrupted etanercept therapy. METHODS: All patients received uninterrupted etanercept 50 mg twice weekly during the first 12 weeks, followed by either continuous (n = 1272) or interrupted (n = 1274) etanercept 50 mg once weekly in the next 12 weeks. The primary effectiveness end point was the proportion of responders (those who achieved a Physician's Global Assessment [PGA] score -70 mV>-20 mV. Results of experiments using mutant NR1 and NR2 subunits identified residues that influence block by CPCn. The inhibition by CPCn was not altered significantly in the mutants at the critical asparagines in the M2 loop, NR1 N616, NR2B N615 and NR2B N616, these residues are known to form the narrowest region of the channel and the binding site of Mg(2+). However, mutations at NR1 N650, located in the vestibule of channel pore, and NR1 D669, located in the extracellular region, reduced the inhibition by CPCn, suggesting that these amino acid residues interact with CPCn. These results suggest that CPCn interacts directly with the mouth or vestibule of the ion channel, like a lid. PMID- 17113197 TI - Transcription of estrogen receptor alpha and beta in mouse cerebral cortex: effect of age, sex, 17beta-estradiol and testosterone. AB - Estrogen actions are mainly mediated by estrogen receptor (ER)alpha and ERbeta which in turn are regulated by several factors including age, sex and gonadal steroid hormones 17beta-estradiol and testosterone. In the present study, we have used nuclear run-off assay to examine the effect of these factors on the rate of transcription of ERalpha and ERbeta of mouse cerebral cortex. The run-off assay result was further corroborated with the measurement of steady state level of ERalpha and ERbeta mRNA by semiquantitative RT-PCR method. Our results reveal that ERalpha transcription rate decreases in old mice of both sexes, whereas ERbeta transcription rate decreases only in old females when compared to their adult counterparts. 17beta-Estradiol supplementation reduces the transcription rate of ERalpha and ERbeta in all groups except in adult male while testosterone treatment down regulates the transcription rate of ERalpha and ERbeta in all groups. The semiquantitative RT-PCR analysis reveals that the level of ERalpha mRNA decreases in old male but shows no effect in old female as compared to adult counterpart. In contrast, ERbeta transcript level decreases in old mice of both sexes. Furthermore, ERalpha mRNA level is higher in adult female than in adult male but no sex-dependent difference is seen in ERbeta mRNA level. Supplementation of 17beta-estradiol shows no significant alteration but testosterone reduces the ERalpha level in male mice, while 17beta-estradiol and testosterone down regulate the ERalpha level in female mice of both ages. In case of ERbeta, 17beta-estradiol decreases the transcript level in all groups except adult male while testosterone treatment results in the down regulation of transcript level in all groups. Thus these findings suggest differential effects of age, sex, 17beta-estradiol and testosterone supplementation on the transcription of mouse ER genes which may account for differences in the protein levels of ERalpha and ERbeta and their functions in the brain. PMID- 17113198 TI - Absence of angiogenic genes modification in Italian ALS patients. AB - To investigate the role of vascular endothelial growth factor (VEGF) and angiogenin (ANG) as genetic determinants in the susceptibility to sporadic ALS in Italian patients. VEGF genotype and haplotype analysis revealed no association between any variants and the risk of ALS. Regarding ANG gene, no mutation was detected and the rs11701 polymorphism, previously described as associated with ALS, was not differently distributed between patients and controls. Overall, our data argue against the hypothesis of both genes as risk factors for motoneuron neurodegeneration, at least in an Italian population. PMID- 17113199 TI - Bioecological control of inflammatory bowel disease. AB - It is today generally accepted, that the intestinal bacterial flora is deeply involved in the pathogenesis of human inflammatory bowel diseases (IBDs), although the exact presence of unwanted or lack of specific crucial bacteria are not yet known. Westerners lack to large extent important immunomodulatory and fibre-fermenting lactic acid bacteria (LAB), bacteria which are present in all with a more primitive rural lifestyle. Acute reduction of flora is observed in disease, including IBD, as well as in mental and physical stress. Some observations suggest the mucosa has lost its ability of holding back the pathogenic flora and prevent close contacts between resident microflora and the epithelial surface. Among the manifestations of IBD are increased inflammation and coagulability, impaired cellular membrane function, exaggerated nitric oxide production and impaired short-chain fatty acid production. Animal studies suggest, in addition to reduced flora, an intimate association with immunostimulatory DNA, malfunctioning trifoil factors, increased splanchnic metabolism and reduced availability of natural antioxidants. Treatment with plant fibres, antioxidants and sometimes probiotics have had limited success. The most dramatic effects are seen in the few cases where total faecal replacement (TFR) has been tried. The general experience this far is that the best effects are obtained with compositions of probiotics rather than with single LAB treatments. PMID- 17113200 TI - Head-to-head comparison on the immunogenicity of two HIV/AIDS vaccine candidates based on the attenuated poxvirus strains MVA and NYVAC co-expressing in a single locus the HIV-1BX08 gp120 and HIV-1(IIIB) Gag-Pol-Nef proteins of clade B. AB - In this investigation we have generated and defined the immunogenicity of two novel HIV/AIDS vaccine candidates based on the highly attenuated vaccinia virus strains, MVA and NYVAC, efficiently expressing in the same locus (TK) and under the same viral promoter the codon optimized HIV-1 genes encoding gp120 and Gag Pol-Nef antigens of clade B (referred as MVA-B and NYVAC-B). In infected human HeLa cells, gp120 is released from cells and GPN is produced as a polyprotein; NYVAC-B induces severe apoptosis but not MVA-B. The two poxvirus vectors showed genetic stability of the inserts. In BALB/c and in transgenic HHD mice for human HLA-A2 class I, both vectors are efficient immunogens and induced broad cellular immune responses against peptides represented in the four HIV-1 antigens. Some differences were observed in the magnitude and breadth of the immune response in the mouse models. In DNA prime/poxvirus boost protocols, the strongest immune response, as measured by fresh IFN-gamma and IL-2 ELISPOT, was obtained in BALB/c mice boosted with NYVAC-B, while in HHD mice there were no differences between the poxvirus vectors. When the prime/boost was performed with homologous or with combination of poxvirus vectors, the protocols MVA-B/MVA-B and NYVAC-B/NYVAC-B, or the combination NYVAC-B/MVA-B gave the most consistent broader immune response in both mouse models, although the magnitude of the overall response was higher for the DNA-B/poxvirus-B regime. All of the immunization protocols induced some humoral response against the gp160 protein from HIV-1 clone LAV. Our findings indicate that MVA-B and NYVAC-B meet the criteria to be potentially useful vaccine candidates against HIV/AIDS. PMID- 17113201 TI - Efficient protein boosting after plasmid DNA or recombinant adenovirus immunization with HIV-1 vaccine constructs. AB - DNA plasmids and recombinant adenovirus serotype-5 (rAd5) vectors are being studied in human clinical trials as HIV-1 vaccine candidates. Each elicits robust T-cell responses and modest antibody levels. Since protein immunization alone elicits antibody but not CD8 T-cell responses, we studied protein boosting of DNA and rAd5 HIV-1 vaccine vectors. A single Env protein immunization provided a marked boost in antibody titer in guinea pigs primed with either DNA or rAd5 vaccines, and the resulting antibody binding and neutralization levels were similar to those attained after thee sequential protein immunizations. Since both T-cell immunity and neutralizing antibodies are thought to be required for protection against HIV-1, it may be possible to establish a balanced T-cell and antibody response with appropriate vectored vaccines and improve the neutralizing antibody titer with protein boosting. PMID- 17113202 TI - FLK-1-based minigene vaccines induce T cell-mediated suppression of angiogenesis and tumor protective immunity in syngeneic BALB/c mice. AB - Angiogenesis is a rate-limiting step in the development of tumors. Here, we demonstrate that oral minigene DNA vaccines against murine vascular endothelial growth factor receptor-2 (FLK-1), a self-antigen overexpressed on proliferating endothelial cells in the tumor vasculature, induced protection against tumors of different origin in syngeneic BALB/c mice. This protection is mediated by CD8 T cells, which specifically kill FLK-1(+) endothelial cells, resulting in marked suppression of tumor angiogenesis. More importantly, the minigene vaccine proved to be of similar efficacy as a vaccine encoding the whole FLK-1 gene. These data suggest a FLK-1 minigene vaccine provides a more flexible alternative to the whole gene vaccine and will facilitate their future design and clinical applications in cancer therapy and prevention. PMID- 17113203 TI - Market incentives, human lives, and AIDS vaccines. AB - For many, an AIDS vaccine holds the promise of intervening in a widespread epidemic because it is not predicated on changing economic structures and social contexts underlying vulnerability to HIV for millions of individuals. Yet 20 years into the AIDS epidemic, there is still no vaccine. Based on interviews of AIDS vaccine researchers, watchdog organizations, and ethics groups from the United States, South Africa, and Kenya conducted between August and December of 2003, this paper explores possible answers to the question of why there is no vaccine, looking in particular at contradictions between a biomedical research industry increasingly driven by market incentives and a disease that primarily affects individuals living in low-income countries with little vaccine purchasing power. Producing a vaccine that could be effective in low-income regions requires new kinds of initiatives that can coordinate research nationally and globally, and circumvent current regulatory mechanisms that dictate against the development and dissemination of low-profit medical technologies. Until such initiatives are supported, however, vaccine research will continue at a devastatingly slow pace at the cost of millions of lives annually. PMID- 17113204 TI - Looking for answers: eye movements in non-visual cognitive tasks. AB - It is not known why people move their eyes when engaged in non-visual cognition. The current study tested the hypothesis that differences in saccadic eye movement rate (EMR) during non-visual cognitive tasks reflect different requirements for searching long-term memory. Participants performed non-visual tasks requiring relatively low or high long-term memory retrieval while eye movements were recorded. In three experiments, EMR was substantially lower for low-retrieval than for high-retrieval tasks, including in an eyes closed condition in Experiment 3. Neither visual imagery nor between-task difficulty was related to EMR, although there was some evidence for a minor effect of within-task difficulty. Comparison of task-related EMRs to EMR during a no-task waiting period suggests that eye movements may be suppressed or activated depending on task requirements. We discuss a number of possible interpretations of saccadic eye movements during non-visual cognition and propose an evolutionary model that links these eye movements to memory search through an elaboration of circuitry involved in visual perception. PMID- 17113205 TI - Categorization and affect: evidence for intra-hemispheric interactions. AB - Both emotional reactivity and categorization have long been studied within the framework of hemispheric asymmetry. However, little attempt has been made to integrate both research areas using any form of neuropsychological research, despite behavioral data suggesting a consistent relationship between affective and categorization processes. The primary goal of the current study was to examine the possibility of a laterally mediated interaction between emotional reactivity and the cognitive process of categorization. Using a split visual fields categorization task combined with affect inducing procedures, we hypothesized that the relationship between state affect and categorization would be dependent on the nature of state affect and on the hemisphere targeted. Results offered support for this hypothesis, showing that state affect related changes in categorization appeared only in the hemisphere commonly associated with both a specific affective state and categorization strategy employed. Findings are discussed in terms of possible evidence for a hemispheric arousal effect underlying the relationship between affect and categorization. PMID- 17113206 TI - Assessment of malondialdehyde levels in schizophrenia: a meta-analysis and some methodological considerations. AB - Oxidative stress has long been suggested to participate in the pathophysiology of schizophrenia. Most of the published data on the subject rely on malondialdehyde levels assessment through thiobarbituric acid reactive substances (TBARS), despite significant methodological concerns. The present paper reports on a meta analysis of studies published up to July 2006. Relevant studies were identified through PubMed search and the References section of experimental and review papers on the subject. Studies were retained for analysis if they provided adequate methods description, controls and subjects numbers, means and standard deviations. Using a random effect model, an effect size of 1.22 (CI: 0.64-1.80) was found, with significant bias and high heterogeneity. Inspection of the bias assessment plot suggested a lower value in the order of d approximately 0.5, in line with the results of the larger studies. The heterogeneity of the results appeared to be unrelated to assay type or biological sample. However a meta regression analysis suggested an association, at trend level, between the precision-weighted effect size and the proportion of drug-free patients, which might therefore account for some of the heterogeneity. PMID- 17113207 TI - An open-label trial of N-acetylcysteine for the treatment of cocaine dependence: a pilot study. AB - Recent preclinical studies implicate N-acetylcysteine (NAC), a cysteine prodrug, as a potential medication for preventing relapse to cocaine use; however, little is known about the safety and tolerability of NAC in cocaine-dependent subjects in an outpatient setting. This pilot study examines the safety and tolerability of 3 doses of NAC for the treatment of cocaine dependence. Twenty three treatment seeking cocaine-dependent patients participated in a 4-week medication trial and received NAC at doses of 1200 mg/day, 2400 mg/day or 3600 mg/day. Results suggested that the three doses were well tolerated. Overall, the retention rates appeared to favor higher doses of NAC (2400 mg/day and 3600 mg/day). The majority of subjects who completed the study (n=16) either terminated use of cocaine completely or significantly reduced their use of cocaine during treatment. Overall the findings suggest that it is feasible to treat cocaine-dependent treatment seekers with N-acetylcysteine on an outpatient basis. PMID- 17113208 TI - Assessment of CPR-D skills of nurses in Goteborg, Sweden and Espoo, Finland: teaching leadership makes a difference. AB - INTRODUCTION: Construction of an effective in-hospital resuscitation programme is challenging. To document and analyse resuscitation skills assessment must provide reliable data. Benchmarking with a hospital having documented excellent results of in-hospital resuscitation is beneficial. The purpose of this study was to assess the resuscitation skills to facilitate construction of an educational programme. MATERIALS AND METHODS: Nurses working in a university hospital Jorvi, Espoo (n=110), Finland and Sahlgrenska University Hospital, Goteborg (n=40), Sweden were compared. The nurses were trained in the same way in both hospitals except for the defining and teaching of leadership applied in Sahlgrenska. Jorvi nurses are not trained to be, nor do they act as, leaders in a resuscitation situation. Their cardiopulmonary resuscitation (CPR) skills using an automated external defibrillator (AED) were assessed using Objective Structured Clinical Examination (OSCE) which was build up as a case of cardiac arrest with ventricular fibrillation (VF) as the initial rhythm. The subjects were tested in pairs, each pair alone. Group-working skills were registered. RESULTS: All Sahlgrenska nurses, but only 49% of Jorvi nurses, were able to defibrillate. Seventy percent of the nurses working in the Sahlgrenska hospital (mean score 35/49) and 27% of the nurses in Jorvi (mean score 26/49) would have passed the OSCE test. Statistically significant differences were found in activating the alarm (P<0.001), activating the AED without delay (P<0.01), setting the lower defibrillation electrode correctly (P<0.001) and using the correct resuscitation technique (P<0.05). The group-working skills of Sahlgrenska nurses were also significantly better than those of Jorvi nurses. CONCLUSIONS: Assessment of CPR-D skills gave valuable information for further education in both hospitals. Defining and teaching leadership seems to improve resuscitation performance. PMID- 17113209 TI - Outcomes from out-of-hospital cardiac arrest in Detroit. AB - OBJECTIVE: To determine the out-of-hospital cardiac arrest survival rate, and prevalence of modifiable factors associated with survival, in Detroit, Michigan, over a 6-month period of time in 2002. METHODS: A retrospective review of all out of-hospital cardiac arrests responded to by the Detroit Fire Department, Division of Emergency Medical Services. All elements of the EMS runsheet were transcribed to a database for analysis. Patient hospital records were reviewed to determine survival to hospital admission. All survivors to hospital admission were surveyed later in the Michigan Department of Vital Records death registry search. RESULTS: During this study timeframe, there were 538 confirmed out-of-hospital cardiac arrests within the City of Detroit, of which 67 were excluded for being dead on scene [51 (12.5%)] or having no available hospital records [16 (3.0%)]. Of the remaining 471 patients, 443 (94.1%) died before hospital admission. Only 44 (9.9%) of the 471 patients had a first recorded rhythm of ventricular fibrillation (VF), and 339 (76.5%) were asystolic. Of the 28 patients who survived to hospital admission, only 2 (7.1%) were noted to have a first rhythm of VF, and 15 (53.6%) were asystolic. Only one patient survived to hospital discharge. CONCLUSIONS: In this urban setting, out-of-hospital cardiac arrest is an almost uniformly fatal event. PMID- 17113210 TI - Treatment of polycystic ovary syndrome with spironolactone plus licorice. AB - OBJECTIVE: The aim of the study was to compare the effect of spironolactone (antagonist of mineralocorticoid and androgen receptors) versus spironolactone plus licorice (agonist of mineralocorticoid receptors and mild inhibitor of androgen synthesis) on plasma renin activity, aldosterone and androgen levels in women with polycystic ovary syndrome (PCOS). STUDY DESIGN: Thirty-two women with PCOS were divided into two groups: 16 received 100 mg spironolactone and 16 spironolactone plus 3.5 g of licorice a day. Blood pressure, body mass index, serum electrolytes, plasma renin activity, plasma aldosterone and cortisol, serum testosterone, and urinary tetrahydrocortisol/tetrahydrocortisone ratio were measured before and during treatment. RESULTS: Mean blood pressure was significantly reduced during spironolactone treatment, while it was unchanged in women receiving spironolactone plus licorice. Twenty percent of women treated with spironolactone and none treated with the addition of licorice complained of symptoms related to volume depletion. Consistently, the activation of the renin aldosterone system was significantly lower during spironolactone plus licorice than with spironolactone alone. The prevalence of metrorrhagia was lower in the combined therapy. CONCLUSIONS: In patients with PCOS the mineralocorticoid properties of licorice can reduce the prevalence of side effects related to the diuretic activity of spironolactone. PMID- 17113211 TI - Effect of latency period after premature rupture of membranes on 2 years infant mortality (DOMINOS study). AB - OBJECTIVE: To assess the impact of a short latency period after preterm premature rupture of the membranes (PPROM) on infant mortality. STUDY DESIGN: A prospective cohort study of women with PPROM between 24(0/7) and 33(6/7) weeks' in singleton gestation was performed in all maternity wards of the Rhone-Alpes Region. Neonatal and infant outcomes were compared according to the latency period (<48 h and > or =48 h). The primary outcome was the mortality rate and the secondary outcome was a composite variable of significant neurological disorders at 2 years of age. Outcomes was stratified according to gestational age at rupture. Univariate and multiple logistic regression analyses were used with SAS statistical software. RESULTS: Out of 471 women recruited in the study at a mean gestational age of 30.5+/-0.2 weeks, 170 (37%) presented with a <48-h latency period, and 301 (63%), a > or =48-h latency period. While prior to 30 weeks' gestation, the mortality rate was higher in neonates with a short latency period (16.3% versus 7.3%, p < 0.01) with pulmonary disease being the major cause of death, a short latency period was associated with a lower mortality rate after 30 weeks' gestation (0% versus 3.7%, p=0.02). After adjusting for confounding factors, a <48-h latency period remained an independent factor associated with infant mortality prior to 30 week's gestation (odds ratio 3.8, 95% confidence interval 1.3-11.7). Significant neurological disorders were not modified by the length of the latency period. CONCLUSION: For PPROM that occur before 30 weeks' gestation, a short latency period was associated with a higher infant mortality rate. Inversely, it was associated with a lower mortality rate after 30 weeks'. There is an urgent need for a thorough evaluation of expectant management of PPROM after 30 weeks' gestation. PMID- 17113212 TI - Twin ectopic pregnancy in a previous cesarean scar section and subsequent fertility. PMID- 17113213 TI - The role of magnetic resonance imaging in the local staging of penile cancer. AB - OBJECTIVES: To assess the accuracy of magnetic resonance imaging (MRI) as a local staging technique in penile cancer and its role in selecting patients for conservative surgical management. METHODS: Fifty-five men diagnosed with invasive penile carcinoma on biopsy were locally staged with the use of MRI. Prostaglandin E1 (alprostadil) was injected into the corpora to induce an artificial erection. Radiologic staging was compared against final histopathologic stage of the tumour. Sensitivity, specificity, and kappa agreement values were calculated for each tumour stage. Additionally, corpora cavernosa involvement was reviewed in 20 consecutive cases and suitability for conservative surgery assessed. RESULTS: A good correlation between radiologic and histologic staging was achieved with an overall kappa value of 0.75 (p<0.001). Stage-specific sensitivities and specificities were calculated: T1 (85%; 83%), T2 (75%; 89%), and T3 (88%; 98%). MRI accurately predicted corpora cavernosa invasion in all cases of pathologically proven disease. These patients were selected to undergo partial penectomy. There were no complications using this imaging technique. CONCLUSIONS: This study demonstrates that penile MRI is highly accurate in the local staging of penile cancer. Associated improvements in surgical planning allow the provision of conservative surgical treatments over more radical procedures. PMID- 17113214 TI - Predicting the outcome of prostatectomy using noninvasive bladder pressure and urine flow measurements. AB - OBJECTIVES: To determine whether categorisation of bladder outlet obstruction (BOO) using measurements of bladder pressure and urine flow obtained by a novel noninvasive medical device (the penile cuff test) improves prediction of outcome from endoscopic prostatectomy (TURP). METHODS: A consecutive cohort of 208 men undergoing TURP following standard assessment in our institution was recruited, and 179 (86%) completed the protocol. Each subject underwent a penile cuff test prior to surgery; outcome was assessed by change in IPSS at 4 mo. The proportion of men with good outcome (>50% reduction in IPSS) was compared according to categorisation by noninvasive bladder pressure and urine flow measurements. RESULTS: The cuff test was completed by 93% of men with 2% experiencing an adverse event. Men categorised as having BOO by the test (37% of total) had an 87% chance of a good outcome from TURP (p<0.01), whilst of those deemed not obstructed (19% of total) 56% experienced good outcome (p<0.01). For the remaining men not categorised in these two groups, 77% had good outcome, which was identical to the result of the cohort as a whole (77%, p=NS). CONCLUSIONS: Urodynamic categorisation using measurements obtained by the noninvasive penile cuff test improves prediction of outcome for men with LUTS undergoing TURP. This finding together with the ease and acceptability of the test suggest its suitability for office-based clinical use to assist men and their physicians in the selection for surgical treatment for relief of LUTS. PMID- 17113215 TI - Histologic subtype of metastatic renal cell carcinoma predicts response to combined immunochemotherapy with interleukin 2, interferon alpha and 5 fluorouracil. AB - OBJECTIVES: Combined immunochemotherapy with interleukin 2 (IL-2), interferon alpha (IFN-alpha), and 5-fluorouracil (5-FU) is an established first-line therapy for metastatic renal cell carcinoma (RCC). However, data on histologic parameters predictive of clinical benefit are rare. In this study, we evaluated the response to immunochemotherapy in the main histologic subtypes of renal cell carcinoma and performed a subgroup analysis of inoperable patients. METHODS: From 164 patients treated with one or two cycles of combined immunochemotherapy, radical nephrectomy had revealed 22 cases of papillary RCC (pRCC; 13.4%) and 131 cases of clear cell RCC (ccRCC; 79.9%). In the remaining 11 (6.7%) their disease was inoperable. The overall response rates were evaluated according to World Health Organization criteria. RESULTS: For ccRCC and inoperable disease, responses of 34.4% and 27.3% after one cycle and 28.8% and 16.7% after two cycles, respectively, were noted. In contrast, no patient with pRCC showed any response after two cycles of combined immunochemotherapy. CONCLUSIONS: No objective response was seen in patients with pRCC. Hence, the use of immunotherapeutic agents must be questioned in this histologic subtype. PMID- 17113216 TI - Sacral neuromodulation for refractory lower urinary tract dysfunction: results of a nationwide registry in Switzerland. AB - OBJECTIVE: To assess the efficacy and safety of sacral neuromodulation (SNM) in patients with refractory lower urinary tract dysfunction in Switzerland based on a nationwide registry. PATIENTS AND METHODS: A total of 209 patients (181 females, 28 males) underwent SNM testing between July 2000 and December 2005 in Switzerland. Subjective symptom improvement, bladder/pain diary variables, adverse events, and their management were prospectively registered. RESULTS: SNM testing was successful (defined as improvement of more than 50% in bladder/pain diary variables) in 102 of 209 patients (49%). An implantable pulse generator (IPG) was placed in 91 patients (89% of all successfully tested and 44% of all tested patients). Of the IPG-implanted patients, 71 had urge incontinence, 13 nonobstructive chronic urinary retention, and 7 chronic pelvic pain syndrome. After a median follow-up of 24 mo, SNM was successful in 64 of the 91 IPG implanted patients (70%) but failed in 27 patients. SNM was continued in 15 of the 27 patients considered failures, because following troubleshooting SNM response improved subjectively and the patients were satisfied. However, improvement in bladder/pain diary variables remained less than 50%. In the other 12 patients both the leads and the IPG were explanted. During the test phase and during/following IPG implantation, 6% (12 of 209) and 11% (10 of 91) adverse event rates and 1% (3 of 209) and 7% (6 of 91) surgical revision rates were reported, respectively. CONCLUSIONS: SNM is an effective and safe treatment for refractory lower urinary tract dysfunction. Adverse events are usually transient and can be treated effectively. PMID- 17113217 TI - Implications of prostate-specific antigen doubling time as indicator of failure after surgery or radiation therapy for prostate cancer. AB - OBJECTIVES: To review the methodology of PSA doubling time (PSA DT) calculations and the implications of PSA DT for the follow-up of prostate cancer patients curatively treated with surgery or radiation therapy. METHODS: A literature search of the most recent articles on PSA DT (those published after 2000) led to the selection of six studies with the largest and best-documented cohorts of patients treated with surgery or irradiation with curative intent. RESULTS: PSA kinetics, in the form of PSA DT, is the most effective parameter for identifying patients at significant risk for mortality specific to prostate cancer. Thresholds of 3, 6, and 12 mo have shown prognostic significance both in surgical and radiation series, notwithstanding differences in treatment selection, definition of biochemical recurrence, and methods of DT calculation. CONCLUSIONS: In retrospective studies, PSA DT is a reliable predictor of prognosis; however, prospective validation studies are needed to determine the cut points of PSA DT. Optimal time intervals for calculation and optimal thresholds are still to be determined. PMID- 17113218 TI - Re: Giacomo Novara, Antonio Galfano, Vincenzo Ficarra and Walter Artibani. Anticholinergic drugs in patients with bladder outlet obstruction and lower urinary tract symptoms: a systematic review. Eur urol 2006;50:675-83. PMID- 17113219 TI - Endothelial nitric oxide synthase keeps erection regulatory function balance in the penis. AB - OBJECTIVES: We evaluated the regulatory influence of endothelial nitric oxide (NO) on the basal functional states of the NO and RhoA/Rho-kinase signaling pathways in the penis using endothelial NO synthase (eNOS) mutant mice and eNOS gene transfer technology. METHODS: Four groups of mice were used: wild type (WT), eNOS gene deleted (eNOS-/-), eNOS and neuronal NOS gene deleted (dNOS-/-), and eNOS-/- mutant mice transfected intracavernosally with eNOS. Cyclic guanosine monophosphate (cGMP) concentration, protein kinase G (PKG) activity, activated RhoA, and Rho-kinase activity were determined in penes of WT and both mutant mouse groups. Constitutive NOS and PKG activities, RhoA, Rho-kinase-alpha and beta isoforms, and phosphorylated myosin light-chain phosphatase target subunit (p-MYPT-1) expressions and Rho-kinase activity were determined in penes of eNOS-/ mice after eNOS gene transfer. RESULTS: Compared with results in the WT penis, eNOS-/- and dNOS-/- mutant mouse penes had significant reductions in NOS activity, cGMP concentration, PKG activity, Rho-kinase activity, and p-MYPT-1 expression (p<0.05) with no significant changes in activated RhoA or in RhoA and Rho-kinase-alpha and -beta protein expressions. After eNOS gene transfer to penes of eNOS-/- mice, Rho-kinase-beta and p-MYPT-1 expressions and total Rho-kinase activity were significantly increased from baseline levels (p<0.05). CONCLUSIONS: These data suggest that endothelial NO has a role in the penis as a regulator of the basal signaling functions of the NO and RhoA/Rho-kinase erection mediatory pathways. These data offer new insight into the homeostasis of erection regulatory biology. PMID- 17113220 TI - Bioturbation-induced phosphorous release from an insoluble phosphate source. AB - The influence of bioturbation caused by common carp fry in 5 L jars (5 L each) in the laboratory and in 150 L outdoor vats in increasing the fertilizer value of phosphate rock was evaluated. Soluble reactive phosphate (SRP) was determined to quantify the effects of bioturbation, fish excrements and soil. The level of SRP was always lowest in the control series. Introduction of common carp fry resulted in a net increase of 0.09-0.10 mg L(-1) of SRP attributable to the effect of fish excrement. Bioturbation caused by common carp resulted in a 64.8-90% influx of phosphate from bottom soil in the presence of phosphate rock but only about 6.3 7.2% in the absence of phosphate rock. The bioturbation that occurred in these treatments resulted in a significant release of phosphorous into the overlying water from the apatite source. The results confirm the benefits of the application of environmentally friendly phosphate rock in fish farming ponds at low cost. PMID- 17113221 TI - Standardization of FSH, LH and hCG--current position and future prospects. AB - Gonadotropin measurements contribute significantly to patient management in both endocrinology and oncology. Differences in calibration, antibody specificities and assay design mean that gonadotropin results obtained in different methods are still not comparable. Comparing patient results obtained in different methods therefore remains problematic, whether for individual patient care, when assessing the results of multicentre clinical trials, or when formulating national and international guidelines and recommendations. Achieving improved comparability of results for these important analytes will require clear descriptive nomenclature, accurate calibration with highly purified standards, careful characterization of what gonadotropin isoforms methods are measuring, broad recommendations about the most clinically appropriate antibody combinations, and increased awareness of clinically relevant interferences and the action required to minimise their effect. Encouraging manufacturers to standardize and carefully describe the evaluation methods they use, such that data from different manufacturers can readily be compared, is also a pre requisite for future progress. PMID- 17113222 TI - p16(INK4a) promoter hypermethylation in oral scrapings of oral squamous cell carcinoma risk patients. AB - Aberrant promoter hypermethylation is common in oral squamous cell carcinoma (OSCC) and may be useful as a marker for cancer cells. The aim of our study was to assess whether the detection of hypermethylation of the promoter region of p16(INK4a) in oral cytological samples could be a marker in patients with risk of developing an OSCC. Hypermethylation was detected in 20% (29/145) of the cases analyzed. These findings provide further evidence that the inactivation of p16(INK4a), due to methylation, is an early event in the carcinogenic pathway of OSCC. PMID- 17113223 TI - Breast cancer risk associated with AURKA 91T -->A polymorphism in relation to BRCA mutations. AB - In this study 759 breast cancer patients, including 9 BRCA1 and 98 BRCA2 mutation carriers, and 653 mutation-negative unaffected controls were genotyped for the AURKA 91T -->A polymorphism. Individuals homozygous for the 91A allele were found to be at increased risk of breast cancer compared to 91T homozygotes (OR=1.87; 95% CI=1.09-3.21). This association was strengthened when cases carrying BRCA mutations were excluded (OR=2.00; 95% CI=1.15-3.47). BRCA carrier cases differed from sporadic cases and their allele distribution was very similar to controls. These results show a statistically significant increased risk of sporadic breast cancer for individuals that are homozygous for the 91A allele but no effect in carriers of BRCA mutations. This may throw light on previously conflicting results. PMID- 17113224 TI - Dietary intake of B-vitamins, polymorphisms in thymidylate synthase and serine hydroxymethyltransferase 1, and colorectal adenoma risk: a Dutch case-control study. AB - Thymidylate synthase and serine hydroxymethyltransferase are involved in folate metabolism. In a case-control study, including 768 cases and 709 controls, we investigated the associations between colorectal adenomas and TS tandem repeat and SHMT1 C1420T polymorphisms, and the interplay with B-vitamins. The polymorphisms were not associated with adenomas, but there was a borderline significant interaction between TS genotype and vitamin B6: the association between vitamin B6 and adenomas seemed positive in TS 3R/3R individuals, but inverse in TS 2R/2R individuals. This study does not provide evidence for a role of SHMT1 genotype in adenoma occurrence. Future research has to indicate whether the TS-B6 interplay is a real effect or a chance finding. PMID- 17113225 TI - The boiling liquid expanding vapour explosion (BLEVE): mechanism, consequence assessment, management. AB - Among the most devastating of accidents likely in chemical process industry is the boiling liquid expanding vapour explosion (BLEVE). It is accompanied by highly destructive blast waves and missiles. In most situations there is also a fireball or a toxic gas cloud. The damaging effect of BLEVEs is reflected in the fact that the 80-odd major BLEVEs that have occurred between 1940 and 2005 have claimed over a 1000 lives and have injured over 10,000 persons besides harming property worth billions of dollars. Release of toxic chemicals like chlorine and phosgene from BLEVEs have damaged large chunks of areas surrounding the BLEVE site. This paper presents an overview of the mechanism, the causes, the consequences, and the preventive strategies associated with BLEVEs. PMID- 17113226 TI - Adsorption of a cationic dye (methylene blue) onto spent activated clay. AB - The adsorption characteristics of methylene blue (MB) onto spent activated clay (SAC), a waste produced from an edible oil manufacturer was investigated. Results showed that the adsorption increased with increasing MB concentration, temperature, and pH. The adsorption equilibrium data was well fitted by multilayer adsorption isotherm. The maximum adsorption capacities for MB ranged from 0.94x10(-4) to 3.41x10(-4)mol/g between 5 and 45 degrees C. Thermodynamic parameters suggest that the adsorption is spontaneous and endothermic. We proposed a modified double exponential equation accounting both with chemical and mathematical point of view to describe the adsorption kinetic data. The increases of mass transfer and adsorption capacity were mainly attributed to the interlayer of the SAC expanding at higher temperature. An activation energy of 13.5 kcal/Kmol was determined suggesting that the adsorption involved a chemical reaction mechanism. PMID- 17113227 TI - Arsenic removal by electrocoagulation using combined Al-Fe electrode system and characterization of products. AB - Combination of electrodes, such as aluminum and iron in a single electrochemical cell provide an alternative method for removal of arsenic from water by electrocoagulation. The removal process has been studied with a wide range of arsenic concentration (1-1000 ppm) at different pH (4-10). Analysis of the electrochemically generated by-products by XRD, XPS, SEM/EDAX, FT-IR, and Mossbauer Spectroscopy revealed the expected crystalline iron oxides (magnetite (Fe3O4), lepidocrocite (FeO(OH)), iron oxide (FeO)) and aluminum oxides (bayerite (Al(OH)3), diaspore (AlO(OH)), mansfieldite (AlAsO(4).2(H2O)), as well as some interaction between the two phases. The amorphous or very fine particular phase was also found in the floc. The substitution of Fe3+ ions by Al3+ ions in the solid surface has been observed, indicating an alternative removal mechanism of arsenic in these metal hydroxides and oxyhydroxides by providing larger surface area for arsenic adsorption via retarding the crystalline formation of iron oxides. PMID- 17113228 TI - Evaluation of gas removal and bacterial community diversity in a biofilter developed to treat composting exhaust gases. AB - The performance of a new, but simply constructed, biofilter system, developed to purify composting exhaust air, was evaluated. The biofilter was packed with mature compost mixed with activated carbon and sludge sourced from a wastewater treatment plant. An alternating air flow system and a bioaerosol reduction device were designed to prevent pressure drop and reduce bioaerosol release. Experimental results demonstrated that satisfactory removal efficiencies of nitrogen-containing compounds, sulfur-containing compounds, fatty acids, total hydrocarbon and odor were achieved at an empty bed retention time (EBRT) of 30s. No significant acidification or alkalinity in the biofilter was observed, and the system was characterized by a small pressure drop and a low level of bioaerosol emission. Denaturing gradient gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH) techniques were used to uncover the changes in the bacterial community of the biofilter during the deodorization processes. A minimum of 16 bands were observed in the DGGE profile. Phylogenetic analysis revealed the phylum of Proteobacteria to be predominant, followed by Actinobacteria, Bacteroidetes, and Firmicutes, in descending order. However, the occurrence and predominance of specific bacterial species varied with the environmental conditions of the biofilter. Our results demonstrate - from both an engineering and biological point of view - the feasibility of the biofilter system described herein in purifying the gases derived from composting food waste. PMID- 17113229 TI - The influence of different temperature programmes on the bioremediation of polycyclic aromatic hydrocarbons (PAHs) in a coal-tar contaminated soil by in vessel composting. AB - The biodegradation of 16 US. EPA-listed polycyclic aromatic hydrocarbons (sigma PAHs), with accompanying humification and microbial community structure changes during simulated in-vessel composting-bioremediation of an aged coal-tar contaminated soil amended with green waste were studied over 56 days. The experimental design compared one constant temperature profile (TC=38 degrees C) with three variable temperature profiles (TP1, TP2 and TP3), including treatment at 70 degrees C to comply with regulatory requirements. Greatest sigma PAHs removal (75.4+/-0.1%; k(1)=0.026 day(-1), R(2)=0.98) occurred at TC=38 degrees C compared to all variable temperature profiles TP1 (62.1+/-11.0%; k(1)=0.016 day( 1), R(2)=0.93), TP2 (71.8+/-8.2%; k(1)=0.021 day(-1), R(2)=0.95) and TP3 (45.3+/ 9.7%; k(1)=0.010 day(-1), R(2)=0.91). This study proved that using thermophilic temperatures (70 degrees C) towards the end of in-vessel composting processes (TP2) resulted in greater sigma PAHs removal than using other variable temperature profiles (TP1, TP3), as long as the increase was stepwise via an intermediate temperature (55 degrees C). Phospholipid fatty acid (PLFA) signatures indicated that use of thermophilic temperatures towards the end of the in-vessel composting-bioremediation (TP2) resulted in a higher fungal to bacterial PLFA ratio and a lower Gram-positive to Gram-negative (G(+)/G(-)) bacterial ratio. Fluorescence excitation-emission matrix (EEM) showed the presence of peaks typical of humic-like (Ex/Em wavelength pair approximately 340/460 nm) and fulvic-acid-like (Ex/Em wavelength pair approximately 245/460 nm) substances, indicating mineralization and/or maturation of the compost. Varying the temperature during in-vessel composting to comply with regulatory requirements for pathogen control, promoted contaminant biodegradation, microbial activity and compost maturation. PMID- 17113230 TI - Protection of vincristine-induced neuropathy by WldS expression and the independence of the activity of Nmnat1. AB - The slow Wallerian degeneration protein (WldS), a fusion protein containing amino terminal E4B and full-length nicotinamide mononucleotide adenylyltransferase 1 (Nmnat1), delays axon degeneration caused by physical damages, toxins and genetic mutations which result in patients being diagnosed with neurodegenerative diseases. It is still controversial whether the suppression of axonal degeneration by WldS is due to Nmnat1 or other portion. We generated WldS or Nmnat1-overexpressing Neuro2A cell lines, in which neuronal differentiation including neurite elongation can be induced by retinoic acid. The overexpression of WldS delayed the neurite degeneration by vincristine, whereas that of Nmnat1 did not delay it much. Taken together, Nmnat1 is considerably weaker than WldS for protection from toxic injury in vitro, suggesting that amino-terminal region of WldS is likely to be more significant for protection from axonal degeneration. PMID- 17113231 TI - Association of APOE with Parkinson disease age-at-onset in women. AB - APOE polymorphism has received extensive attention as a risk factor for Parkinson's disease (PD), but findings have been equivocal. Analysis of APOE variants in an Australian PD case-control sample revealed a robust association between genotype and age-at-onset (AAO) of PD in women (P=0.0008). These data not only further implicate APOE in PD, but also provide a stark example of the effects that gender may play in complex disorders. PMID- 17113232 TI - Doramectin and albendazole resistance in sheep in The Netherlands. AB - A faecal egg count reduction test was conducted on a sheep farm with suspected avermectin resistance. Five groups of 10 sheep were formed. Group 1 was the untreated control group. Groups 2-5 were treated according to weight with the recommended dose of, respectively, levamisole, doramectin, moxidectin or albendazole. Resistance was found in the sheep treated with doramectin (15% efficacy) and albendazole (87% efficacy). Levamisole and moxidectin were 100 and 99% effective, respectively. Larval identification of the faecal cultures of the doramectin-treated sheep revealed 100% Haemonchus contortus larvae. After albendazole treatment, 77% of the cultured larvae were H. contortus and 23% Teladorsagia/Trichostrongylus. Because there is a lot of trade in sheep on the farm, it is probable that the resistant worms were introduced with livestock from other farms than being selected on the farm. PMID- 17113233 TI - Involvement of cytoplasmic membrane damage in the copper (II)-dependent cytotoxicity of a novel naturally occurring tripyrrole. AB - In the presence of a nonlethal concentration of Cu(II), washed Escherichia coli ATCC8739 cells were killed by a novel tripyrrole 1, isolated as a red pigment from the Serratia sp. Cell killing was accompanied by a depletion in the potassium pools of the cells due to the damage to the cytoplasmic membrane, without any detectable DNA damage as revealed by the transformed plasmid DNA and phage induction assay. This revealed that the bactericidal activity of compound 1 in the presence of Cu(II) results from membrane damage. Induction of endogenous catalase in the E. coli cells increased their resistance against the combination of compound 1 and Cu(II). Although compound 1 alone generated large amount of reactive oxygen species (ROS), it did not show any cell killing against E. coli in the absence of Cu(II). The Cu(II)-dependent bactericidal activity of compound 1 was suppressed by ethylenediaminetetraacetate, bathocuproine, catalase and superoxide disumutase (SOD), but not by dimethyl sulfoxide. These findings suggest that recycling redox reactions between Cu(II) and Cu(I), involving compound 1 and hydrogen peroxide on the cell surface, must be important in the mechanism of the killing. Compound 1 alone showed selective bactericidal activity against the gram positive bacterium, Bacillus cereus ATCC 6630, possibly due to its differential cellular transport. PMID- 17113234 TI - The 17q12-q21 amplicon: Her2 and topoisomerase-IIalpha and their importance to the biology of solid tumours. AB - Her2 and topoisomerase-IIalpha (T2A) gene amplification are separate events, although the latter is more frequently seen in Her2 amplified (34-90%) than in Her2 non-amplified (5-10%) tumours. There is a better correlation between Her2 amplification and protein overexpression in breast cancer (BC) than in other tumour types. This marker is also considered a powerful prognostic factor in BC, with similar data emerging in other solid tumours such as bladder, ovarian, endometrial, gastro-oesophageal and non-small cell lung cancer. Her2 amplification and/or overexpression are highly predictive of response to HER2 targeted compounds such as trastuzumab and lapatinib but have been inconsistent predictors of response to cytotoxic chemotherapy. There is also evidence that these tumours are relatively resistant to anti-oestrogen therapy (tamoxifen) but not to oestrogen deprivation (e.g. with aromatase inhibitors). T2A aberrations are uncommon events in solid tumours, with an overall prevalence of approximately 10%. T2A amplification has shown inconsistent correlation with T2A protein expression in preclinical and clinical studies, mainly because non-genetic events such as proliferation rate can also affect protein expression. Expression of T2A protein has not been shown to reliably predict response to T2A inhibitors, despite the fact that this enzyme is the direct target for these compounds. In BC, T2A amplification appears to be a good predictor of response to anthracyclines, but these data are still in the process of validation. The significance of T2A deletions is currently under investigation, but contrary to what was previously thought, it may also predict benefit from treatment with T2A inhibitors. The prognostic significance of T2A aberrations is currently unknown. PMID- 17113235 TI - Human autoantibodies against early endosome antigen-1 enhance excitatory synaptic transmission. AB - Early endosome antigen 1 (EEA1), a peripheral membrane protein associated with the cytoplasmic face of early endosomes, controls vesicle fusion during endocytosis, as extensively studied in non-neuronal cells. In neurons, early endosomes are involved in recycling of synaptic vesicles and neurotransmitter receptors. Since certain patients bearing autoantibodies that target EEA1 develop neurological disease, we studied the subcellular distribution of EEA1 in neurons and the effect on neurotransmission of purified immunoglobulins from the serum of a patient bearing EEA1 autoantibodies. EEA1 was localized in the soma and in the postsynaptic nerve terminals. Electrophysiological recordings in hippocampal slices including purified EEA1 antibodies in the patch pipette solution, revealed a run-up of AMPA, N-methyl-D-aspartate and kainate receptor-mediated excitatory post-synaptic currents recorded from CA3 pyramidal neurons, which was absent in the recordings obtained in the presence of control human immunoglobulin G. Inclusion of human EEA1 antibodies had no effect on inhibitory post-synaptic responses. Recordings in the presence of a dominant-negative C-terminal EEA1 deletion mutant produced a similar effect as observed with human anti-EEA1 antibodies. This specific effect on the excitatory synaptic transmission may be due to the impairment of internalization of specific glutamate receptors and their subsequent accumulation in the synapse. These results may account for the neurological deficits observed in some patients developing EEA1 autoantibodies. PMID- 17113236 TI - Rembrandt's Maria Bockenolle has a butterfly rash and digital deformities: overlapping syndrome of rheumatoid arthritis and systemic lupus erythematosus. AB - Rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) are the most common autoimmune disorders, although they each have very different pathophysiology. In general, RA is considered to be a Th1-mediated disease, while SLE is a Th2-mediated disease. Thus, their overlapping, in so called "rhupus", is a rare condition. In Rembrandt van Rijn's (1606-1669) portrait of the middle-aged Maria Bockenolle, we have what may be the earliest depiction of a case of rhupus syndrome: the coexistence of a butterfly rash and digital deformities. This suggests the possible historical importance of an RA epidemic which took place in the early 17th century. PMID- 17113237 TI - Alzheimer's disease Braak Stage progressions: reexamined and redefined as Borrelia infection transmission through neural circuits. AB - Brain structure in health is a dynamic energized equation incorporating chemistry, neuronal structure, and circuitry components. The chemistry "piece" is represented by multiple neurotransmitters such as Acetylcholine, Serotonin, and Dopamine. The neuronal structure "piece" incorporates synapses and their connections. And finally circuits of neurons establish "architectural blueprints" of anatomic wiring diagrams of the higher order of brain neuron organizations. In Alzheimer's disease, there are progressive losses in all of these components. Brain structure crumbles. The deterioration in Alzheimer's is ordered, reproducible, and stepwise. Drs. Braak and Braak have described stages in the Alzheimer disease continuum. "Progressions" through Braak Stages benchmark "Regressions" in Cognitive function. Under the microscope, the Stages of Braak commence in brain regions near to the hippocampus, and over time, like a tsunami wave of destruction, overturn healthy brain regions, with neurofibrillary tangle damaged neurons "marching" through the temporal lobe, neocortex and occipital cortex. In effect the destruction ascends from the limbic regions to progressively destroy the higher brain centers. Rabies infection also "begins low and finishes high" in its wave of destruction of brain tissue. Herpes Zoster infections offer the paradigm of clinical latency of infection inside of nerves before the "marching commences". Varicella Zoster virus enters neurons in the pediatric years. Dormant virus remains inside the neurons for 50-80 years, tissue damage late in life (shingles) demonstrates the "march of the infection" down neural pathways (dermatomes) as linear areas of painful blisters loaded with virus from a childhood infection. Amalgamation of Zoster with Rabies models produces a hybrid model to explain all of the Braak Stages of Alzheimer's disease under a new paradigm, namely "Alzheimer's neuroborreliosis" in which latent Borrelia infections ascend neural circuits through the hippocampus to the higher brain centers, creating a trail of neurofibrillary tangle injured neurons in neural circuits of cholinergic neurons by transsynaptic transmission of infection from nerve to nerve. PMID- 17113238 TI - Some fibrocystic breast change may be caused by sexually transmitted H. pylori during oral nipple contact: supporting literature and case report of resolution after gut H. pylori eradication treatment. AB - OBJECTIVES: To briefly review previously published evidence for Helicobacter pylori (Hp), colonization of extra-intestinal sites and suggest an hypothesis that breast acini and ducts be added to this list, concluding such breast colonization is not rare and is a sexually transmitted infection. METHODS: PubMed literature search and review with a case report. CONCLUSIONS: (1) Evidence indicates oral Hp is common and can remain in the mouth after successful eradication in stomach and duodenum. (2) Evidence indicates that the breast is also occasionally colonized by Hp. (3) Hp may be injected retrograde up into ducts of the breast during oral nipple stimulation during sexual activity and this Hp may give rise to some cases of fibrocystic breast change. (4) A case of painful fibrocystic change that had been present for two years in a 27 year old female, resolved after gastrointestinal Hp treatment. PMID- 17113239 TI - Targeting SERCA2a as an innovative approach to the therapy of congestive heart failure. AB - CHF prevalence is continuously increasing worldwide and maintains one of the poorest prognoses of any major disease. Abundant evidence points to derangement of Ca(2+) cycling as the primary biochemical mark of the failing myocyte. Istaroxime is a novel compound with a dual mechanism of action: inhibition of Na(+), K(+)-ATPase and stimulation of SERCA2a. The increase in cytoplasmic Ca(2+) due to Na(+), K(+)-ATPase inhibition together with greater sarcoplasmic reticulum reloading result in both increased inotropy and lusitropy. This effect is seen in normal and failing in vitro and in vivo models. Istaroxime improvement of the contraction-relaxation cycle constitutes a novel therapeutic approach to the treatment of heart failure. PMID- 17113240 TI - Effectiveness of complete diagnostic examination in clinical practice settings. AB - BACKGROUND: Thorough follow-up of a positive fecal occult blood test (FOBT) result, or a complete diagnostic evaluation (CDE), is recommended as routine care on the basis of findings from colorectal cancer (CRC) screening trials. CDE involves either colonoscopy or the combination of flexible sigmoidoscopy and double contrast barium enema X-ray. However, little evidence outside clinical screening trial settings has been reported in the literature to support CDE performance. The focus of this study was to determine the impact of CDE in primary care practice settings. METHODS: We determined diagnostic outcomes for 461 adult patients with a positive FOBT result in 318 primary care practices in southeastern Pennsylvania and southern New Jersey. Sociodemographic data were collected and CDE status was ascertained for these patients. Polytomous logistic models were used to identify whether having CDE was associated with subsequently being diagnosed with lower gastrointestinal "neoplastic disease" or "other gastrointestinal disease" as compared to "normal findings. RESULTS: Patients who underwent CDE were significantly more likely to have a reported diagnosis of colorectal neoplasia than normal findings (adjusted odds ratio = 3.65, 95% confidence interval = 1.58-8.39, p = 0.02). CDE performance did not result in the differential diagnosis of other gastrointestinal disease. CONCLUSIONS: Patients with a positive screening FOBT who underwent CDE were more likely to be diagnosed with colorectal neoplasia than with less serious conditions or have normal findings. Results support the use of CDE in CRC screening. PMID- 17113241 TI - Nucleophosmin and human cancer. AB - Nucleophosmin (NPM) is a nucleolar phosphoprotein that shuttles between the nucleus and cytoplasm during the cell cycle. NPM has several interacting partners and diverse cellular functions, including the processing of ribosomal RNA, centrosome duplication and the control of cellular processes to ensure genomic stability. Subcellular localization of NPM appears to be strongly correlated with NPM functions and cell proliferation. NPM is phosphorylated mainly at its central acidic domain by several upstream kinases, and its phosphorylation appears to be involved in regulating its functions in ribosome biogenesis and centrosome duplication. Recent studies suggest that NPM may act as a licensing factor to maintain proper centrosome duplication and that the Ran/CRM1 nucleocytoplasmic complex regulates local trafficking of NPM to centrosomes by interacting through its nuclear export sequence motif. Here, we provide a brief overview of NPM functions and its roles in human carcinogenesis, and discuss our recent findings related to the potential mechanisms underlying its regulation of centrosome duplication. PMID- 17113242 TI - Novel adenine adducts, N7-guanine-AFB1 adducts, and p53 mutations in patients with schistosomiasis and aflatoxin exposure. AB - INTRODUCTION: The most frequent mutation in human hepatocellular carcinoma (HCC) in populations exposed to a high dietary intake of aflatoxin B1 (AFB1) is a mutation in codon 249 of the p53 gene. Schistosomiasis is known to cause p53 mutation. We hypothesized that the combination of schistosomiasis and aflatoxin B1 increases the incidence of p53 gene mutation. METHODS: Liver tissue from 21 patients with schistosomiasis and 5 patients without schistosomiasis were analyzed for occurrence of mutations of the p53 gene and levels of N7-guanine AFB1 adducts. RESULTS: The presence of mutations in codon 249 of p53 gene was higher in patients infected with Schistosoma haematobium (S. haematobium) than in those infected with Schistosoma mansoni (S. mansoni) or a combination of both strains (p < 0.01), compared to control subjects. No mutations were detected in p53 gene in liver DNA from schistosomiasis-free patients. Significant amounts of N7-guanine-AFB1 adducts and novel adenine-adducts (p < 0.01) were detected in patients with schistosomiasis, mostly in patients infected with S. haematobium or a combination of both strains, compared to control subjects. CONCLUSION: These data suggest that schistosomiasis and exposure to aflatoxin B1 act synergistically to increase the incidence of p53 gene mutation. The increase in p53 mutations may enhance progression of HCC at an early age in patients with schistosomiasis. PMID- 17113243 TI - Further investigation of psychological and environmental correlates of substance use in adolescence in six European countries. AB - AIM: To study the multifactorial correlates of adolescents' use of legal and illegal substances in six European countries and to assess whether a common pattern of factors exists irrespective of the countries' different sociocultural backgrounds. DESIGN: Cross-sectional European school population survey (ESPAD) following standardized methodology. PARTICIPANTS: National probability samples of 16-year-old high school students from Bulgaria, Croatia, Greece, Romania, Slovenia and the UK. Total sample 16,445. MEASUREMENTS: Anonymous questionnaire self-administered in the classroom. Self-reported use of tobacco, alcohol, marijuana and other illegal drugs. Correlates examined: environmental, such as peer culture and family-related; behavior-related such as antisocial behavior, truancy and anomie; and psychological factors such as self-esteem and depressive mood. FINDINGS: Separate logistic regressions for the two genders produced a set of psychosocial correlates common to the use of all legal and illegal substances. The strongest were peer and older sibling models of use, and peer-oriented lifestyle, followed by patterns of antisocial behavior and truancy. Family related variables such as not living with both parents, parental monitoring and relationships with parents were less significant. Self-esteem and depressive mood were not significant. Girls' use of substances, especially illegal ones, showed stronger associations than boys' with a deviant behavior pattern. Few interactions between country and other correlates were significant. CONCLUSIONS: Common correlates can be identified across countries. Older siblings' and peers' substance have a strong impact on adolescents' use. Preventive interventions should include all substances with addictive potential. PMID- 17113244 TI - Identification of the Salmonella phage epsilon 34 tailspike gene. AB - To understand the interaction between lipopolysaccharide (LPS) and proteins in molecular detail, a molecular genetic approach has been employed, using phage as a model system. The phage epsilon(34) is a Salmonella phage whose tailspike protein (TSP) uses the host LPS as its initial host cell receptor. Previous studies indicated that there was a similarity between the well-studied tail protein of Salmonella phage P22 and the epsilon(34). This study reports the identification of the gene for the epsilon(34) TSP as well as its initial characterization. In addition, some aspects of the structure of the epsilon(34) TSP have been deduced. PMID- 17113245 TI - Insight in to the phylogeny of polyhydroxyalkanoate biosynthesis: horizontal gene transfer. AB - Polyhydroxyalkanoates (PHAs) are gaining more and more importance the world over due to their structural diversity and close analogy to plastics. Their biodegradability makes them extremely desirable substitutes for synthetic plastics. PHAs are produced in organisms under certain stress conditions. Here, we investigated 253 sequenced (completely and unfinished) genomes for the diversity and phylogenetics of the PHA biosynthesis. Discrepancies in the phylogenetic trees for phaA, phaB and phaC genes of the PHA biosynthesis have led to the suggestion that horizontal gene transfer (HGT) may be a major contributor for its evolution. Twenty four organisms belonging to diverse taxa were found to be involved in HGT. Among these, Bacillus cereus ATCC 14579 and Xanthomonas axonopodis pv. citri str. 306 seem to have acquired all the three genes through HGT events and have not been characterized so far as PHA producers. This study also revealed certain potential organisms such as Streptomyces coelicolor A3(2), Staphylococcus epidermidis ATCC 12228, Brucella suis 1330, Burkholderia sp., DSMZ 9242 and Leptospira interrogans serovar lai str. 56601, which can be transformed into novel PHA producers through recombinant DNA technology. PMID- 17113247 TI - Non-coding RNAs: lost in translation? AB - In the last ten years, several RNAs with no protein-coding potential have been accumulating in RNA databases and are in need of further molecular characterization. At the same time, examples of non-coding RNAs (ncRNAs) such as microRNAs, small RNAs, small interfering RNAs (siRNAs) and medium/large RNAs with various functions have been described in the literature. Recent evidence points to a widespread role of these molecules in eukaryotic cells, suggesting that the majority of the new ncRNA examples might have specific functions. The aim of this review is to describe several new functional ncRNAs that have been recently identified and characterized, providing some clues that these molecules might not be produced by chance or as by-products of transcription as has been speculated. PMID- 17113246 TI - Mitogenomic analysis for coelacanths (Latimeria chalumnae) caught in Tanzania. AB - In recent years, a large number of individuals of the species Latimeria chalumnae, one of the living fossil coelacanths, have been landed off the coast of Tanzania. Although L. chalumnae specimens have also been landed at other localities in the western Indian Ocean, so far, viable populations of this species have been identified only at two localities, Comoros and South Africa. Therefore, the recent active catch off Tanzania suggests a new habitat for L. chalumnae. To examine the genetic background of the Tanzanian fish, we analyzed complete mtDNA sequences of two Tanzanian individuals (Kigombe-9 and Songo Mnara 1) collected from the north and south coasts of Tanzania. Using the recently reported criteria for six haplotypes established in a population genetic study for coelacanths living in the western Indian Ocean [Schartl, M., Hornung, U., Hissman, K., Schauer, J., Fricke, H., 2005. Relatedness among east African coelacanths. Nature 435, 901.], we characterized Songo Mnara-1 as haplotype 1 and Kigombe-9 as haplotype 5. We suggest that the Songo Mnara specimen is a member of the Comoran group, but was swept away by the South Equatorial current. The individual from Kigombe may be a member of an undiscovered population that exists near the boundary between Tanzania and Kenya. Further analysis using more than 19 individuals recently captured off the north coast of Tanzania will reveal whether a new population exists there. Our sequence data suggest additional variable sites in the mtDNA sequence that may define the population structure of coelacanths in the western Indian Ocean and also raise the possibility that the previously published Comoran coelacanth mtDNA sequence contains several critical errors including base changes and indels. PMID- 17113248 TI - The intragenomic polymorphism of a partially inverted repeat (PIR) in Gallus gallus domesticus, potential role of inverted repeats in satellite DNAs evolution. AB - We report here the molecular characterization of the basic repeating unit of a novel repetitive family, partially inverted repeat (PIR), previously identified from chicken genome. This repetitive DNA family shares a close evolutionary relationship with XhoI/EcoRI repeats and chicken nuclear-membrane-associated (CNM) repeat. Sequence analyses reveal the 1430 bp basic repeating unit can be divided into two regions: the central region ( approximately 1000 bp) and the flanking region ( approximately 430 bp). Within the central region, a pair of repeats (86 bp) flanks the central core ( approximately 828 bp) in inversed orientation. Due to the tandem array feature shared by the repeating units, the inverted repeats fall between the central core and flanking region. Southern blot analyses further reveal the intragenomic polymorphism of PIR, and the molecular size of repeating units ranges from 1.1 kb to 1.6 kb. The identified monomer variants may result from multiple crossing-over events, implying the potential roles of inverted repeats in satellite DNAs variation. PMID- 17113249 TI - Highly pathogenic avian influenza virus subtype H5N1 in Mute swans in the Czech Republic. AB - In order to determine the actual prevalence of avian influenza viruses (AIV) in wild birds in the Czech Republic extensive surveillance was carried out between January and April 2006. A total of 2101 samples representing 61 bird species were examined for the presence of influenza A by using PCR, sequencing and cultivation on chicken embryos. AIV subtype H5N1 was detected in 12 Mute swans (Cygnus olor). The viruses were determined as HPAI (highly pathogenic avian influenza) and the hemagglutinin sequence was closely similar to A/mallard/Italy/835/06 and A/turkey/Turkey/1194/05. Following the first H5N1 case, about 300 wild birds representing 33 species were collected from the outbreak region and tested for the presence of AIV without any positive result. This is the first report of highly pathogenic avian influenza subtype H5N1 in the Czech Republic. The potential role of swan as an effective vector of avian influenza virus is also discussed. PMID- 17113251 TI - The liberation of thiocholine from acetylthiocholine (ASCh) by pralidoxime iodide (2=PAM) and other oximes (obidoxime and diacetylmonoxime). PMID- 17113252 TI - Early postnatal exposure to cigarette smoke impairs the antigen-specific T-cell responses in the spleen. AB - Annually, approximately two million babies are exposed to cigarette smoke in utero and postnatally through cigarette smoking of their mothers. Exposure to mainstream cigarette smoke is known to impair both innate and adaptive immunities, and it has been hypothesized that the effects of in utero exposure to cigarette smoke on children's health might primarily stem from the adverse effects of cigarette smoke on the immune system. To simulate the environment that babies from smoking mothers encounter, we examined the effects of prenatal mainstream and postnatal sidestream cigarette smoke on spleen cell responses. Results show that postnatal exposure of newborn Balb/c mouse pups to sidestream cigarette smoke through the first 6 weeks of life strongly suppresses the antibody response of spleen cells to the T-cell-dependent antigen, sheep red blood cells. The reduction in the antibody response seen within 6 weeks of postnatal smoke exposure is much quicker than the published data on the time 25 weeks) required to establish reproducible immunosuppression in adult rats and mice. Moreover, the immunosuppression is not associated with significant changes in T-cell numbers or subset distribution. While the postnatal exposure to cigarette smoke did not affect the mitogenic response of T and B cells, the exposure inhibited the T cell receptor-mediated rise in the intracellular calcium concentration. These results suggest that the early postnatal period is highly sensitive to the immunosuppressive effects of environmental tobacco smoke, and the effects are causally associated with impaired antigen-mediated signaling in T cells. PMID- 17113253 TI - Sustained antibacterial effect of a hand rub gel incorporating chlorhexdine loaded nanocapsules (Nanochlorex). AB - In the present study, an original chlorhexidine-loaded nanocapsule-based gel (Nanochlorex) was tested as hand rub gel against the resident skin flora in comparison with 2-propanol 60% (v/v) and 62% (v/v) ethanol-based gel (Purell). After 30-s hand rub, the immediate bactericidal effect of Nanochlorex was found comparable to 2-propanol 60% (v/v) (reduction factor, RF: 0.30+/-0.35 versus 0.38+/-0.55, P>0.05) against aerobic bacteria, whereas the post-values of surviving anaerobes were shown significantly lower from Nanochlorex (P<0.001) and insignificant from 2-propanol 60% (v/v) (P>0.05). Sustained antibacterial effect of Nanochlorex was confirmed against the resident and transient hand flora in two sets of experiment. In the first, the results obtained with the glove-juice technique showed that the bactericidal effect induced by Nanochlorex hand rub persisted throughout 3-h period, while Purell failed to reduce significantly the post-values of surviving bacteria. In the second, repeated artificial contaminations with Staphylococcus epidermidis was carried out onto ex vivo human skin pre-treated by either Nanochlorex or Purell for 5min, then maintained in cell diffusion apparatus for 4h. The log(10) reduction of surviving bacteria was significantly higher with Nanochlorex than that determined with Purell after three successive contaminations (from approximately 5.5 to 1.5 log(10) reduction for Nanochlorex between the first and the third contamination; approximately 1log(10) reduction for Purell throughout the experiment), confirming the sustained antibacterial effect of chlorhexidine-loaded nanocapsule-based gel. The immediate and sustained antibacterial effect of Nanochlorex was explained by chlorhexidine carrier system which improved the drug targeting to bacteria and reduced from osmotic gel further bacterial growth on the skin. Nanochlorex) might constitute a promising approach for hygienic hand disinfection in care practice performing multiple procedures. PMID- 17113254 TI - Mechanisms of cisplatin-induced ototoxicity and prevention. AB - Cisplatin is a widely used chemotherapeutic agent to treat malignant disease. Unfortunately, ototoxicity occurs in a large percentage of patients treated with higher dose regimens. In animal studies and in human temporal bone investigations, several areas of the cochlea are damaged, including outer hair cells in the basal turn, spiral ganglion cells and the stria vascularis, resulting in hearing impairment. The mechanisms appear to involve the production of reactive oxygen species (ROS), which can trigger cell death. Approaches to chemoprevention include the administration of antioxidants to protect against ROS at an early stage in the ototoxic pathways and the application of agents that act further downstream in the cell death cascade to prevent apoptosis and hearing loss. This review summarizes recent data that shed new light on the mechanisms of cisplatin ototoxicity and its prevention. PMID- 17113255 TI - In vitro study of the effects of khat (Catha edulis Forsk) extract on isolated mouse interstitial cells. AB - Isolated mouse interstitial cells were incubated with different concentrations of khat (Catha edulis) extract (0.06 mg/ml, 0.6 mg/ml, 6 mg/ml, 30 mg/ml and 60 mg/ml) and cell viability as well as testosterone concentration measured at 30 min intervals over a 3h incubation period. High concentrations of khat extract (30 mg/ml and 60 mg/ml) significantly inhibited testosterone production while low concentrations (0.06 mg/ml, 0.6 mg/ml and 6 mg/ml) significantly stimulated (P<0.05) testosterone production by mouse interstitial cells. Similarly, at concentrations of 30 mg/ml and 60 mg/ml, there was a significant decrease in interstitial cell viability, whereas at 0.06 mg/ml, 0.6 mg/ml and 6 mg/ml there was no significant decrease. There was only a weak correlation (r=0.39) between testosterone production and viable interstitial cells. We postulate that khat extract at high concentrations may cause reproductive function impairment in the user but at low concentrations, may enhance testosterone production with accompanying effects on reproductive functions in male mice. PMID- 17113256 TI - Kaempferia parviflora ethanolic extract promoted nitric oxide production in human umbilical vein endothelial cells. AB - The rhizomes of Kaempferia parviflora (KP) (Zingiberaceae) have been used in Thai traditional medicine for health promotion and for the treatment of digestive disorders and gastric ulcer. This study investigated effect of KP on endothelial function. Studies in human umbilical vein endothelial cells (HUVEC) showed that KP dose-dependently increased nitrite concentrations in culture media after 48 h incubation. eNOS mRNA and protein expression were also enhanced. The induction of eNOS mRNA was detected at 4 h and plateau at 48 h while iNOS expression was not observed. These data demonstrate that KP has a great potential for a supplemental use in vascular endothelial health promotion. PMID- 17113257 TI - Drugs held and sold by pharmacists of the Jewish community of medieval (11-14th centuries) Cairo according to lists of materia medica found at the Taylor Schechter Genizah collection, Cambridge. AB - The importance of the Genizah for the research of the medieval Mediterranean communities, supplying information on almost every aspect of life, is well known among historian. Less known is that pharmacy was the most popular of all branches of the healing art in the medieval Jewish community of Cairo, according to the Genizah manuscripts. Sources for study of medieval practical drugs are extremely rare since most records naturally vanish over the years, and only some medical books, which contained theoretical pharmacology, have survived to the present day. Drugs lists enable us to understand medieval practical pharmacy and to reconstruct their inventories. This study reports on 71 original drugs lists that were found in the Genizah; they are different from merchants' letters dealing with commerce in drugs and give no instructions for the use or preparation of formulas as usually found in prescriptions. Twenty-six lists are written in Judeo Arabic and 45 in Arabic, none of the lists is written in Hebrew. The longest list contains 63 identified substances. These lists were apparently used by pharmacists for professional and business purposes as inventories of drugs, records, orders, or even receipts. Two hundred and six different drugs are mentioned in the drugs lists of which 167 are of plant origin, 16 are of animal origin, and the remaining 23 are inorganic. The lists point directly to the place they occupied on the shelves of the pharmacies that could be found in the lanes and alleys of the Jewish quarter of Cairo. The most frequently mentioned substance were myrobalan (27), pepper and saffron (21), lentisk (15), almond, basil, rose, rosemary (14), cattle products, camphor and spikenard (13). PMID- 17113258 TI - Effects of pre- and neonatal exposure to bisphenol A on murine brain development. AB - Bisphenol A (BPA), known as an environmental endocrine disrupter, is widely used in industry and dentistry. We investigated the effects of fetal and neonatal exposure to bisphenol A (BPA) on the brain development of mice. The density of tyrosine hydroxylase (TH)-immunoreactive (IR) neurons in substantia nigra was significantly decreased in BPA-exposed female mice (3 microg/g powder food), but not in the male mice, as compared with that of the control mice. The densities of calbindin D-28 K-, calretinin- and parvalbumin-IR neurons in the cerebral cortex were not different between BPA-exposed and the control mice. The present study indicates that chronic exposure of BPA during prenatal and neonatal periods causes a decrease of TH-positive neurons in substantia nigra only in female mice brain. PMID- 17113259 TI - Clinical application of high and ultra high-field MRI. AB - The 21st century ushered in the century of human beings. The previous era characterized by the chase after super technology has been replaced by a new era which focuses on the meaning of human existence and quality of life. Clinical practice has accordingly also undergone rapid change. Amongst the many competing technologies, clearly magnetic resonance technology, especially ultra high-field magnetic resonance imaging, plays a major role in defining current clinical practice. Elimination of all invasive aspects from diagnostic imaging, including intravenous infusion or use of ionizing radiation, is one of the final goals of the new generation of clinical imaging. This goal is especially worthwhile when one consider the welfare of children. Technological MRI advancements are steadily bridging the gap towards this goal. With T2 reversed and three-dimensional anisotropy (3DAC) contrast imaging on a 3.0T system, the anatomical resolution of routine clinical images has reached a level of resolution equivalent to general pathology. Realistic imaging microscopy application is also on the horizon with the establishment of clinical 7.0T systems. Individual brain activation maps can now be readily obtained under clinical settings thanks to high-field functional MRI (fMRI). Nevertheless, because active self-organizing processes of cortical functionalities are under active development in the pediatric population, fMRI has only limited, if any, clinical usage in children. Similarly, whereas connectivity analysis in the individual patient using diffusion tensor imaging (DTI) has little clinical usage in the pediatric population, DTI can be successfully applied to multiple subject analysis for exploring unknown connectivity abnormalities in this age group. Magnetic resonance spectroscopy (MRS) and its pictorial display (spectroscopic imaging) is now finding more and more clinical applications across the age spectrum of patients. PMID- 17113260 TI - MR spectroscopy and diffusion tensor imaging of the brain in congenital muscular dystrophy with merosin deficiency: metabolite level decreases, fractional anisotropy decreases, and apparent diffusion coefficient increases in the white matter. AB - Brain magnetic resonance spectroscopy (MRS) and diffusion tensor imaging (DTI) in one patient with merosin-deficient congenital muscular dystrophy (MDCMD) revealed significant metabolite (choline, creatine, N-acetyl aspartate) level reductions, fractional anisotropy (FA) reduction and increased apparent diffusion coefficient (ADC) in the white matter (p<0.01, all). In the gray matter, the MRS properties did not differ significantly from those in controls. The ADC and FA, however, differed significantly as in the white matter, although the differences were less pronounced. This is the first quantitative MR study of the brain in a patient with MDCMD, which revealed that the concentrations of all MRS measured metabolites were decreased only in the white matter. This observation, combined with the DTI observed ADC increases and FA decrease, indicated a presence of vasogenic edema in the white matter. PMID- 17113261 TI - EEG abnormalities in West syndrome: correlation with the emergence of autistic features. AB - Autism may develop in children with West syndrome. This study was conducted to determine if EEG abnormalities in patients with West syndrome predict the later onset of autism. Two groups of patients with West syndrome, older than 6 years of age, were studied. One group consisted of those with a past history of West syndrome plus autism (N=14); the control group consisted of those with a past history of West syndrome but without autism (N=14). Patients were followed at regular intervals and video-EEG recordings were done. A total of 108 (autistic group) and 123 (non-autistic group) video-EEGs were examined. The two groups were compared with respect to age, presence or absence of hypsarrhythmia, and characteristics and localization of the epileptogenic foci. chi2 and Fisher's exact tests were used. The number of patients with at least one hypsarrhythmic EEG at the age of one year or later was significantly higher in autistic subjects (86%) than in non-autistic controls (29%). The incidence of EEGs with hypsarrhythmia was also higher in the autistic group, especially in older children (autistic, 49% versus non-autistic, 18% at age 3 years and later). Frontal predominance of the primary foci on EEGs with or without hypsarrhythmia was seen in 95.3% of the autistic group and 28.8% of the non-autistic group (p=0.001). Frontal abnormalities on the EEGs, which were mainly bilateral, and the persistence of hypsarrhythmia were significantly related to the emergence of autistic behavior in patients with West syndrome. These findings suggest that paroxysmal discharges in the cortical areas undergoing rapid maturation may be involved in the development of autistic features. PMID- 17113262 TI - A cross-sectional test of the similar-trajectory hypothesis among adults with mental retardation. AB - The similar-sequence and the similar-structure hypotheses are the two mainstays of the developmental approach to mental retardation. In the present study, a third way, the similar-trajectory hypothesis, is described and illustrated using the WAIS-R results of adults with and without mental retardation aged from 20 to 54 years. The whole sample (N=633) comprised 306 participants with mental retardation and 327 without mental retardation. Hierarchical regression analyses comparing the two groups showed similar evolutions of scores with increasing age for verbal and performance scales. These results seem to validate the similar trajectory hypothesis, at least for the present samples and for the aspects of cognitive development considered here. Some weaknesses and implications of the study are considered in the discussion. PMID- 17113263 TI - A learning algorithm for adaptive canonical correlation analysis of several data sets. AB - Canonical correlation analysis (CCA) is a classical tool in statistical analysis to find the projections that maximize the correlation between two data sets. In this work we propose a generalization of CCA to several data sets, which is shown to be equivalent to the classical maximum variance (MAXVAR) generalization proposed by Kettenring. The reformulation of this generalization as a set of coupled least squares regression problems is exploited to develop a neural structure for CCA. In particular, the proposed CCA model is a two layer feedforward neural network with lateral connections in the output layer to achieve the simultaneous extraction of all the CCA eigenvectors through deflation. The CCA neural model is trained using a recursive least squares (RLS) algorithm. Finally, the convergence of the proposed learning rule is proved by means of stochastic approximation techniques and their performance is analyzed through simulations. PMID- 17113264 TI - Combinatorial activation of FAK and AKT by transforming growth factor-beta1 confers an anoikis-resistant phenotype to myofibroblasts. AB - Transforming growth factor-beta (TGF-beta) is a prototypical tumour-suppressor cytokine with cytostatic and pro-apoptotic effects on most target cells; however, mechanisms of its pro-survival/anti-apoptotic signalling in certain cell types and contexts remain unclear. In human lung fibroblasts, TGF-beta1 is known to induce myofibroblast differentiation in association with the delayed activation of focal adhesion kinase (FAK) and protein kinase B (PKB/AKT). Here, we demonstrate that FAK and AKT are independently regulated by early activation of SMAD3 and p38 MAPK, respectively. Pharmacologic or genetic approaches that disrupt SMAD3 signalling block TGF-beta1-induced activation of FAK, but not AKT; in contrast, disruption of early p38 MAPK signalling abrogates AKT activation, but does not alter FAK activation. TGF-beta1 is able to activate AKT in cells expressing mutant FAK or in cells treated with an RGD-containing peptide that interferes with integrin signalling, inhibits FAK activation and induces anoikis (apoptosis induced by loss of adhesion signalling). TGF-beta1 protects myofibroblasts from anoikis, in part, by activation of the PI3K-AKT pathway. Thus, TGF-beta1 co-ordinately and independently activates the FAK and AKT protein kinase pathways to confer an anoikis-resistant phenotype to myofibroblasts. Activation of these pro-survival/anti-anoikis pathways in myofibroblasts likely contributes to essential roles of TGF-beta1 in tissue fibrosis and tumour promotion. PMID- 17113265 TI - Sphingosine 1-phosphate phosphatase 2 is induced during inflammatory responses. AB - Sphingosine 1-phosphate (S1P) levels in cells and, consequently, its bioactivity as a signalling molecule are controlled by the action of enzymes responsible for its synthesis and degradation. In the present report, we examined alterations in expression patterns of enzymes involved in S1P-metabolism (sphingosine kinases including their splice variants, sphingosine 1-phosphate phosphatases, and sphingosine 1-phosphate lyase) under certain inflammatory conditions. We found that sphingosine kinase type 1 (SPHK1) mRNA could be triggered in a cell type specific manner; individual SPHK1 splice variants were induced with similar kinetics. Remarkably, expression and activity of S1P phosphatase 2 (SPP2) was found to be highly upregulated by inflammatory stimuli in a variety of cells (e.g., neutrophils, endothelial cells). Bandshift analysis using oligonucleotides spanning predicted NFkappaB sites within the SPP2 promoter and silencing of NFkappaB/RelA via RelA-directed siRNA demonstrated that SPP2 is an NFkappaB dependent gene. Silencing of SPP2 expression in endothelial cells, in turn, led to a marked reduction of TNF-alpha-induced IL-1beta mRNA and protein and to a partial reduction of induced IL-8, suggesting a pro-inflammatory role of SPP2. Notably, up-regulation of SPP2 was detected in samples of lesional skin of patients with psoriasis, an inflammatory skin disease. This study provides detailed insights into the regulation of SPP2 gene expression and suggests that SPP2 might be a novel player in pro-inflammatory signalling. PMID- 17113266 TI - Schneider's first rank symptoms and continuous performance disturbance as indices of dysconnectivity of left- and right-hemispheric components of language in schizophrenia. AB - BACKGROUND: Here we investigate pathophysiological dimensions (language disturbance, negative symptoms, lateralisation and the continuous performance test) in relation to ICD-10 and DSM-IV concepts of diagnosis. METHODS: A total of 32 consecutive psychotic patients with at least one Schneider's first rank symptom (SFRS), 15 depressed patients without SFRS and 17 normal volunteers were assessed with the Clinical Language Disorder Rating Scale (CLANG), SFRS, "pure defect" Huber's basic symptoms (HBS), handedness (Annett's pegboard task), and the A-X Continuous Performance Test (AX-CPT). RESULTS: CLANG total score (an index of severity of language disorder) was correlated with the severity of SFRS, a higher leftward shift of handedness, and poorer performance on AX-CPT. Receiver operating characteristic (ROC) analysis showed that only CLANG and AX-CPT variables had adequate predictive validity in separating cases of ICD-10 schizophrenia from other diagnoses. The logistic regression model predicting the presence of ICD-10 schizophrenia was statistically significant using CLANG and AX CPT variables, but not SFRS or other variables. HBS did not correlate with other variables and did not predict ICD-10 diagnosis. CONCLUSIONS: A cross-sectional diagnosis based on language disturbance and CPT performance yields a diagnostic construct largely overlapping with the ICD-10 definition of schizophrenia. We suggest that Schneider's first rank symptoms (that play a large role in the DSM IV concept) can be considered an index of left hemisphere dysconnectivity for language whereas CPT dysfunction reflects dysconnectivity of the right hemisphere for those remoter ("spatial") associations that are closer to Bleuler's core defect and to the chronicity implicit in the ICD definition. Thus the dimensions of language disturbance in psychosis can be traced to specific cortico-cortical dysconnectivities. PMID- 17113267 TI - Tetrachloroethylene exposure and risk of schizophrenia: offspring of dry cleaners in a population birth cohort, preliminary findings. AB - Tetrachloroethylene is a solvent used in dry cleaning with reported neurotoxic effects. Using proportional hazard methods, we examined the relationship between parental occupation as a dry cleaner and risk for schizophrenia in a prospective population-based cohort of 88,829 offspring born in Jerusalem from 1964 through 1976, followed from birth to age 21-33 years. Of 144 offspring whose parents were dry cleaners, 4 developed schizophrenia. We observed an increased incidence of schizophrenia in offspring of parents who were dry cleaners (RR=3.4, 95% CI, 1.3 9.2, p=0.01). Tetrachloroethylene exposure warrants further investigation as a risk factor for schizophrenia. PMID- 17113268 TI - DRD2 C957T polymorphism interacts with the COMT Val158Met polymorphism in human working memory ability. AB - The C957T polymorphism in the dopamine D2 receptor (DRD2) gene and the Val158Met polymorphism in the Catechol-O-Methyl-Transferase (COMT) gene affect dopamine transmission and have been found to be associated with schizophrenia. Since DRD2 in mice and the COMT gene in humans modulate working memory, we examined the relationship and possible interaction of both polymorphisms to working memory performance in 188 healthy adults. Subjects having the DRD2 C/C allele showed the poorest performance in a word serial position test. Moreover, the effect of the C957T genotype was strengthened when interaction with the COMT Val158Met polymorphism was included in the analysis. We propose that an interaction of the DRD2 C957T and COMT Val158Met may be involved in the generation of some working memory deficits in schizophrenia. PMID- 17113269 TI - Alterations in rdxA and frxA genes and their upstream regions in metronidazole resistant Helicobacter pylori isolates. AB - Metronidazole resistance among Helicobacter pylori strains has been related to alterations in gene products having metronidazole nitroreductase activities. RdxA and FrxA proteins are the two major contributing factors. In this investigation, the rdxA and frxA genes and their upstream regions were analyzed in 19 H. pylori isolates, 8 of which were metronidazole-sensitive (MIC < or = 8 microg/mL) and 11 of which were metronidazole-resistant (MIC > or = 8 microg/mL), as determined by the E-test. Among the metronidazole-resistant isolates, three contained both RdxA and FrxA proteins with premature truncation caused by gene nonsense mutations or frameshift mutations, while three contained only stop mutations in FrxA and two only in RdxA. Substitutions of amino acids occurred in other RdxA (5/6) and FrxA (4/5) proteins from metronidazole resistant isolates as compared with those from metronidazole-sensitive ones. All metronidazole-resistant isolates had alterations in RdxA and/or FrxA proteins. Moreover, the upstream regions (-1 to 35) of rdxA and frxA genes in some metronidazole-resistant isolates varied by nucleotide insertion and/or deletion or substitution. The patterns of variation in both genes and their upstream regions were highly diversified. Alterations in rdxA and frxA genes and their upstream regions may be involved in the development of metronidazole resistance in H. pylori. PMID- 17113270 TI - Importance of amino acid alterations and expression of penicillin-binding protein 5 to ampicillin resistance of Enterococcus faecium in Taiwan. AB - The importance of the amino acid sequence in the C-terminal domain of penicillin binding protein 5 (PBP5) and the levels of PBP5 expression to ampicillin resistance of Taiwan clinical isolates of Enterococcus faecium were studied. Sequence data revealed the existence of 12 amino acid sequence variants within the C-terminal domain of PBP5 in the 33 tested isolates (ampicillin minimum inhibitory concentrations (MICs) 1 mg/L to >256 mg/L). Western blot analyses of the levels of PBP5 showed that, with few exceptions, lower amounts of PBP5 were present in the susceptible strains than in the resistant strains. More importantly, a significant correlation (P=0.004, Fisher's exact test) between the expression of PBP5 and ampicillin resistance was detected. Point mutations in PBP5, including addition of aspartic acid or serine after position 466 and change of methionine to alanine or threonine at position 485, alanine or isoleucine to threonine at position 499 and glutamate to valine at position 629, were found to be significantly associated with ampicillin resistance. A significant correlation was obtained for the combined mutation (alleles 10 and 11), suggesting that combined mutation of PBP5 can be a marker for ampicillin resistance of E. faecium. PMID- 17113271 TI - Dysfunction of amyloid precursor protein signaling in neurons leads to DNA synthesis and apoptosis. AB - The classic neuropathological diagnostic markers for AD are amyloid plaques and neurofibrillary tangles, but their role in the etiology and progression of the disease remains incompletely defined. Research over the last decade has revealed that cell cycle abnormalities also represent a major neuropathological feature of AD. These abnormalities appear very early in the disease process, prior to the appearance of plaques and tangles; and it has been suggested that neuronal cell cycle regulatory failure may be a significant component of the pathogenesis of AD. The amyloid precursor protein (APP) is most commonly known as the source of the beta-amyloid (Abeta) peptides that accumulate in the brains of patients with AD. However, a large body of work supports the idea that APP is also a signaling receptor. Most recently, it has been shown that familial AD (FAD) mutations in APP or simple overexpression of wild type APP cause dysfunction of APP signaling, resulting in initiation of DNA synthesis in neurons and consequent apoptosis. In this article, we review the evidence that APP has the potential to activate aberrant neuronal cell cycle re-entry in AD, and we describe a signal transduction pathway that may mediate this abnormal activation of the cell cycle. PMID- 17113272 TI - BCL9-2 binds Arm/beta-catenin in a Tyr142-independent manner and requires Pygopus for its function in Wg/Wnt signaling. AB - The Wingless (Wg)/Wnt signal transduction pathway controls fundamental processes during animal development. Deregulation of the Wg/Wnt pathway has been causally linked to several forms of cancer, most notably to colorectal cancer. In response to Wg/Wnt signaling, Armadillo/beta-catenin associates in the nucleus with DNA bound TCF and several co-factors, among them Legless/BCL9, which provides a link to Pygopus. Recently, the second vertebrate homologue of Legless, BCL9-2 (or B9L), was characterized and proposed to mediate Wnt signaling in a Pygopus independent manner, by binding to a Tyrosine-142-phosphorylated form of beta catenin. Here we examine the role of Tyrosine-142 phosphorylation in several assays and find that it is neither important for the recruitment of BCL9-2, nor for the transcriptional activity of beta-catenin in cultured mammalian cells, nor in Drosophila for Wg signaling activity in vivo. Furthermore, we demonstrate that BCL9-2 can functionally replace Lgs both in cultured cells as well as in vivo and that this rescue activity depends on the ability of BCL9-2 to bind Pygo. Our results do not show a significant functional difference between BCL9-2 and BCL9 but rather suggest that the two proteins represent evolutionary duplicates of Legless, which have acquired distinct expression patterns while acting in a largely redundant manner. PMID- 17113273 TI - In vitro antidrepanocytary actvity (anti-sickle cell anemia) of some congolese plants. AB - Thirty aqueous and ethanolic extracts from 13 congolese plants were evaluated for their antidrepanocytary activity. Twelve of these plants, Alchornea cordifolia, Afromomum albo violaceum, Annona senegalensis, Cymbopogon densiflorus, Bridelia ferruginea, Ceiba pentandra, Morinda lucida, Hymenocardia acida, Coleus kilimandcharis, Dacryodes edulis, Caloncoba welwithsii, and Vinga unguiculata exhibited significant activities, thus, supporting the claims of the traditional healers and suggesting a possible correlation between the chemical composition of these plants and their uses in traditional medicine. PMID- 17113274 TI - Effects of arachidonic acid, docosahexaenoic acid and prostaglandin E(2) on cell proliferation and morphology of MG-63 and MC3T3-E1 osteoblast-like cells. AB - During bone remodelling bone is resorbed by osteoclasts and replaced again by osteoblasts through the process of bone formation. Clinical trials and in vivo animal studies suggest that specific polyunsaturated fatty acids (PUFAs) might benefit bone health. As the number of functional osteoblasts is important for bone formation the effects of specific PUFAs on in vitro osteoblastic cell proliferation were investigated. Morphological studies were conducted to determine whether exposure of the cells to these agents caused structural damage to the cells thereby yielding invalid results. Results from this study showed that arachidonic acid (AA) and docosahexaenoic acid (DHA) both inhibit cell growth significantly at high concentrations. The anti-mitotic effect of AA is possibly independent of PGE(2) production, as PGE(2) per se had little effect on proliferation. Further study is required to determine whether reduced proliferation due to fatty acids could be due to increased differentiation of osteoblasts to the mature mineralising osteoblastic phenotype. PMID- 17113275 TI - Immune epitope mapping in the post-genomic era: lessons for vaccine development. AB - Technological advances have allowed for the creation of ever more complete maps of targets of immune responses in infectious pathogens. The evidence accumulating from such recent studies points to a broader range of targets recognized than previously expected, in terms of both numbers and characteristics of the targeted antigens. Also, multiple studies report a substantial variation in the targets recognized in different human individuals. These findings are not in conflict with the concept of immunodominance, because there are still only a few targets recognized compared with the multitude of potential targets available in a complex pathogen. However, they raise the question if vaccines, which try to emulate protective natural immune responses, should elicit an equally broad range of responses to efficiently convey protection. PMID- 17113276 TI - Diversity and distribution of saprobic microfungi in leaf litter of an Australian tropical rainforest. AB - The diversity and distribution of microfungal assemblages in leaf litter of a tropical Australian forest was assessed using two methods: (1) cultures were isolated using a particle filtration protocol (wet season 2001), and (2) fruit bodies were observed directly on leaf surfaces following incubation in humid chambers (wet and dry season of 2002). Four tree species were studied using both methods, namely Cryptocarya mackinnoniana (Lauraceae), Elaeocarpus angustifolius (Elaeocarpaceae), Ficus pleurocarpa (Moraceae), and Opisthiolepis heterophylla (Proteaceae). An additional two species, Darlingia ferruginea (Proteaceae) and Ficus destruens (Moraceae), were studied using direct observations. In total, fruiting bodies of 185 microfungal species were recorded on leaf surfaces (31-81 species per tree species), and 419 morphotypes were detected among isolates obtained by particle filtration (111-203 morphotypes per tree species). Although the observed microfungal diversity was higher with the particle filtration protocol, both methods concurred with respect to microfungal distributions. The overlap of microfungal species in pair wise comparisons of tree species was low (14-30%), and only 2 and 3% of microfungal species were observed in leaves of all tree species by particle filtration and by direct observations respectively. Multivariate analysis of data from direct observations confirmed the hypothesis that microfungal assemblages are strongly influenced by host phylogeny and are also affected by seasonal and site factors. The importance of host species in shaping microfungal distributions was also supported by the particle filtration data. Several taxa new to science, as well as some widespread saprotrophs, were detected on only one host. The underlying reasons for this affinity remain unclear, but we hypothesise that a number of factors may be involved such as fungal adaptation to plant secondary metabolites or the presence of a biotrophic phase in the fungus' life cycle. PMID- 17113277 TI - External validity of randomized controlled trials in COPD. AB - BACKGROUND: COPD is a heterogeneous disease comprising a wide range of clinical phenotypes, depending on the degree to which emphysema, chronic bronchitis, reversible bronchospasm and small airways inflammation are present. Not all of these phenotypes may be represented among the subjects included in randomized controlled drug trials (RCTs) in COPD, making it difficult for doctors to know to what extent RCT evidence applies to individual patients. From a respiratory health survey of adults randomly selected from the community, we have estimated the proportion of subjects with COPD who would have been eligible for inclusion in major COPD RCTs. METHODS: A postal survey was sent to 3500 randomly selected individuals aged 25-75 years. Respondents were invited to complete a detailed respiratory questionnaire and pulmonary function tests. Subjects with COPD defined by post-bronchodilator spirometry were assessed against the eligibility criteria of 18 major RCTs cited in the Global Initiative for Chronic Obstructive Lung Disease (GOLD) guidelines. FINDINGS: Of 749 subjects completing the full survey, 117 had COPD. Of these, a median of 5% (range 0-20%) of subjects met inclusion criteria for the major RCTs. Of 55 subjects with COPD receiving treatment, 0-9% (median 5%) met inclusion criteria for the major RCTs. INTERPRETATION: The major COPD RCTs on which the GOLD treatment guidelines are based may have limited external validity. Over 90% of the COPD subjects in the community who were taking medication, did so on the basis of RCTs for which they would not have been eligible. PMID- 17113278 TI - Preserved enzymatic activity of glucose oxidase immobilized on unmodified electrodes for glucose detection. AB - Glucose sensing electrodes have been realized by immobilizing glucose oxidase (GOx) on unmodified edge plane of highly oriented pyrolytic graphite (epHOPG) and the native oxide of heavily doped silicon (SiO2/Si). Both kinds of electrode show direct interfacial electron transfer due to the redox process of the immobilized GOx. The measured formal potential of the redox process agrees with that of the native enzyme, suggesting that the immobilized GOx has retained its enzymatic activity. The electron transfer rates of the GOx immobilized electrode are 2s(-1) for GOx/epHOPG electrode and 7.9s(-1) for GOx/SiO2/Si electrode, which are greater than those for which GOx is immobilized on modified electrodes, probably due to the fact that the enzyme makes direct contact to electrode surface. The preservation of the enzymatic activity of the immobilized GOx has been confirmed by observing the response of the GOx/epHOPG and GOx/SiO2/Si electrodes to glucose with a detection limit of 0.050 mM. The response signals the catalyzed oxidation of glucose and, therefore, confirms that the immobilized GOx retained its enzymatic activity. The properties of the electrode as a glucose sensor are presented. PMID- 17113279 TI - Poly-(3-hexylthiophene) self-assembled monolayer based cholesterol biosensor using surface plasmon resonance technique. AB - Cholesterol oxidase (ChOx) has been covalently immobilized onto 1-fluoro-2-nitro 4-azidobenzene (FNAB) modified poly-(3-hexylthiophene) (P3HT) self-assembled monolayer (SAM) onto gold coated glass plates. These ChOx/FNAB/P3HT/Au bio electrodes have been characterized using contact angle (CA) measurements, UV-vis spectroscopy, electrochemical impedance technique, cyclic voltammetric technique and atomic force microscopic (AFM) technique, respectively. The ChOx/FNAB/P3HT/Au bio-electrodes were utilized for the estimation of cholesterol concentration in standard solutions using surface plasmon resonance (SPR) technique. It is shown that this SPR biosensor has linearity from 50 to 500 mg/dl of cholesterol in solution with detection limit of 50 mg/dl, sensitivity of 1.0 4 m degrees /(mg dl), reusability of around 15 times and a shelf-life of about 10 weeks when stored at 4 degrees C. PMID- 17113280 TI - Facultative or obligate anaerobic bacteria have the potential for multimodality therapy of solid tumours. AB - Recent understanding of the unique pathology of solid tumours has shed light on the difficult and disappointing nature of their clinical treatment. All solid tumours undergo angiogenesis that results in biological changes and adaptive metabolisms, i.e. formation of defective vessels, appearance of hypoxic areas, and emergence of an heterogeneous tumour cell population. This micro-milieu provides a haven for anaerobic bacteria. The strictly anaerobic clostridia have several advantages over other facultative anaerobes such as salmonella or lactic acid-producing, Gram-positive, obligate, anaerobic bifidobacteria. Both pathogenic and non-pathogenic clostridia have been demonstrated to specifically colonise and destroy solid tumours. Early trials of non-pathogenic strains in humans had shown plausible safety. Genetic modifications and adaptation of pathogenic and non-pathogenic strains have further created improved features. However, these manipulations rarely generate strains that resulted in complete tumour control alone. Combined modalities of therapies with chemo and radiation therapies, on the other hand, often perform better, including 'cure' of solid tumours in a high percentage of animals. Considering that clostridia have unlimited capacities for genetic improvement, we predict that designer clostridia forecast a promising future for the development of potent strains for tumour destruction, incorporating mechanisms such as immunotherapy to overcome immune suppression and to elicit strong anti-tumour responses. PMID- 17113281 TI - Consultation in palliative care: the relevance of clarification of problems. AB - This study aims to determine the extent and nature of problems in palliative care that are newly identified in the consultation process and the factors influencing their identification. The consultation process includes clarification of problems mentioned by professionals requesting advice. Data are derived from the standard registration forms of Palliative Care Consultation teams. Multilevel logistic regression analysis was carried out with newly identified problem as dependent variable. Fifty seven percent of problems (n=7854) were newly identified. Most newly identified problems were related to physical and pharmacological problems. If psychosocial/spiritual problems were identified, this occurred in most cases through clarification (70%). Newly identified problems were more likely to be identified in the domain of spiritual and psychosocial problems, in bedside consultations, in requests from clinical physicians, and for patients accommodated in a hospice or hospital. Explicit clarification of problems facilitates the identification and addressing of a more comprehensive and specific scope of problems. PMID- 17113282 TI - Application of response surface methodology to cell immobilization for the production of palatinose. AB - Response surface methodology (RSM), based on multivariate non-linear model, was applied to study the interactions and optimization of the immobilization parameters for cell entrapment, namely alginate concentration, cell loading and bead diameter using Erwinia rhapontici NCPPB 1578 that produced palatinose. ANOVA analysis and statistical parameters calculations showed that RSM could be used effectively to model and improve a complex system like cell immobilization. Palatinose yield was increased by 40%. The maximum yield of 140 mg/ml was achieved in a batch of 1h at alginate concentration of 5% w/v, cell loading of 5 g l(-1) and 2.25 mm bead diameter. Thus, the E. rhapontici NCPPB 1578 immobilization in alginate bead and subsequent palatinose yield was successfully improved by application of RSM technique. PMID- 17113283 TI - Batch removal of chromium(VI) from aqueous solution by Turkish brown coals. AB - The ability of using low-rank Turkish brown coals (Ilgin: BC1, Beysehir: BC2, and Ermenek: BC3) to remove Cr(VI) from aqueous solutions was studied as a function of contact time, solution pH, temperature, concentration of metal solutions and amount of adsorbent. Their sorption properties were compared with the activated carbon from Chemviron (AQ-30). Adsorption of Cr(VI) uptake is in all cases pH dependent showing a maximum at equilibrium pH values between 2.0 and 3.2, depending on the biomaterial, that correspond to initial pH values of 2.3 units for BC1, 3.0 units for BC2 and 3.2 units for BC3 and AQ-30. Batch equilibrium tests showed that the Cr(VI) removal was fitted with Freundlich isotherm and the adsorption reached equilibrium in 80 min. It was proceeding effectively into a short acid pH interval (2.0-3.2) where processes of Cr(VI) sorption are maximized. It was observed that the maximum adsorption capacity of 11.2 mM of Cr(VI)/g for Ilgin (BC1), 12.4 mM of Cr(VI)/g for Beysehir (BC2), 7.4 mM of Cr(VI)/g for Ermenek (BC3) and 6.8 mM of Cr(VI)/g for activated carbon (AQ-30) was achieved at pH of 3.0. The rise in temperature caused a slight decrease in the value of the equilibrium constant (K(c)) for the sorption of Cr(VI) ion. The Cr(VI) sorption capacities of Beysehir and Ilgin brown coals were the same. Ermenek brown coals and activated carbon (AQ-30) showed a similar sorption capacity. PMID- 17113284 TI - Metal tolerance and biosorption potential of filamentous fungi isolated from metal contaminated agricultural soil. AB - Heavy metal analysis of agricultural field soil receiving long-term (>20 years) application of municipal and industrial wastewater showed two- to five-fold accumulation of certain heavy metals as compared to untreated soil. Metal resistant fungi isolated from wastewater-treated soil belonged to genera Aspergillus, Penicillium, Alternaria, Geotrichum, Fusarium, Rhizopus, Monilia and Trichoderma. Minimum inhibitory concentrations (MIC) for Cd, Ni, Cr, Cu, and Co were determined. The MIC ranged from 0.2 to 5 mg ml(-1) for Cd, followed by Ni (0.1-4 mg ml(-1)), Cr (0.3-7 mg ml(-1)), Cu (0.6-9 mg ml(-1)) and for Co (0.1-5 mg ml(-1)) depending on the isolate. Aspergillus and Rhizopus isolates were tested for their metal biosorption potential for Cr and Cd in vitro. Biosorption experiments were conducted with initial metal concentrations of 2, 4, 6 and 8 mM with a contact time of 4 h and wet fungal biomass (1-5 g) at 25 degrees C. Maximum biosorption of Cr and Cd ions was found at 6 mM initial metal concentration. Aspergillus sp.1 accumulated 1.20 mg of Cr and 2.72 mg of Cd per gram of biomass. Accumulation of these two metals by very tolerant Aspergillus sp.2 isolate was at par with relatively less tolerant Aspergillus sp.1 isolate. Rhizopus sp. accumulated 4.33 mg of Cr and 2.72 mg of Cd per g of biomass. The findings indicated promising biosorption of cadmium and chromium by the Rhizopus and Aspergillus spp. from aqueous solution. There is little, if any, correlation between metal tolerance and biosorption properties of the test fungi. PMID- 17113285 TI - Addition of Al and Fe salts during treatment of paper mill effluents to improve activated sludge settlement characteristics. AB - Metal salts, ferrous sulphate and aluminium chloride, were added to laboratory scale activated sludge plant treating paper mill effluents to investigate the effect on settlement characteristics. Before treatment the sludge was filamentous, had stirred sludge volume index (SSVI) values in excess of 300 and was moderately hydrophobic. The use of FeSO4.7H2O took three weeks to reduce the SSVI to 90. Microscopic examination showed that Fe had converted the filamentous flocs into a compact structure. When the iron dosing was stopped, the sludge returned to its bulking state within four weeks. In a subsequent trial, the addition of AlCl3 initially resulted in an improvement of the settlement index but then caused deterioration of the sludge properties. It is possible that aluminium was overdosed and caused charge reversal, increasing the SSVI. PMID- 17113286 TI - Hazelnut husk as a substrate for the cultivation of shiitake mushroom (Lentinula edodes). AB - The possibility of using hazelnut husk (HH) as a new basal ingredient for substrate preparation in Lentinula edodes cultivation was investigated. Some chemical properties of the substrates prepared by HH alone and its mixtures with wheat straw (WS), beech wood-chip (BWC) and wheat bran (WB) in different ratios were compared, and their effects on spawn run time, days to first harvest (earliness), yield and biological efficiency (BE) were determined. The N content of the substrate prepared from HH alone was very high (0.82%), and thus the C:N ratio of substrates decreased with an increase in the rate of HH in the mixtures. Yield and BE in the HH alone substrate was considerably low compared with the controls (80BWC:10WS:10M and 60BWC:20WS:20WB), and decreased with an increase in the rate of HH in the mixtures. However, when the HH content in the mixtures was kept below 50%, the yield was relatively high (50HH:50WS and 50HH:50BWC). Even when the HH content increased to 75% in the mixture, the comparable yield and BE to the controls could be obtained by adding 10% of WB as nutrients (75HH:15WS:10WB and 75HH:15BWC:10WB). The results revealed that HH could be used as a new basal ingredient for substrate preparation in L. edodes cultivation. PMID- 17113287 TI - Influence of an additional 2-amino substituent of the 1-aminoethyl pharmacophore group on the potency of rimantadine against influenza virus A. AB - We examined whether the incorporation of a second amino group into the 1 aminoethyl pharmacophore of rimantadine 2 and into the piperidine pharmacophore of the heterocyclic rimantadine 4 was compatible with anti-influenza virus A activity. The new synthetic molecules are capable of forming two hydrogen bonds within the receptor. We identified molecules 8 and 16, bearing the adamantyl and 1,2-diaminoethyl groups, which are equipotent to rimantadine 2 bearing the adamantyl and 1-aminoethyl pharmacophore groups. Interestingly, diamino compound 16 is a 4-fold more potent inhibitor than its parent monoamino heterocyclic rimantadine 4 propably because of additional hydrogen bonding interactions with the M2 protein receptor. PMID- 17113288 TI - Synthesis of 2'-paclitaxel methyl 2-glucopyranosyl succinate for specific targeted delivery to cancer cells. AB - A novel glucose-conjugated paclitaxel 5 was synthesized using succinic acid as linker between 2'-paclitaxel and methyl 2'-glucopyranose. 5 has not only improved the pharmaceutical properties of paclitaxel, such as solubility and stability, but also enhanced the specific target delivery to MCF-7 cells without the cytotoxicity against normal cells. Therefore, the glucose conjugation may be potentially used in the targeted delivery of other drugs into cells via glucose transporters (GLUTs) for cancer therapy. PMID- 17113290 TI - Identification of the benzodiazepines as a new class of antileishmanial agent. AB - The continual increase in drug resistance; the lack of new chemotherapeutic agents; the toxicity of existing agents and the increasing morbidity with HIV co infection mean the search for new antileishmanial agents has never been more urgent. We have identified the benzodiazepines as a structural class for antileishmanial hit optimisation, and demonstrated that their in vitro activity is comparable with the clinically used drug, sodium stibogluconate, and that the compounds are not toxic to macrophages. PMID- 17113289 TI - Solid-phase synthesis of Stat3 inhibitors incorporating O-carbamoylserine and O carbamoylthreonine as glutamine mimics. AB - O-Carbamoylserine and O-carbamoylthreonine are glutamine analogues that were incorporated into a Stat3 inhibitory peptide to probe the requirements of Gln at the pY+3 position. Fmoc-Ser-NHBn and Fmoc-Thr-NHBn were converted to nitrophenyl carbonates and were attached to Rink resin via a side-chain carbamate linkage. After assembly of the peptide, acid treatment resulted in O-carbamoylserine and O carbamoylthreonine-containing peptides. The order of affinity for Stat3 was Gln > Ser(CONH2) >> Thr(CONH2) suggesting a relatively tight binding pocket for the side chain of glutamine. PMID- 17113291 TI - Synthesis and DNA-binding ability of C2R-fluoro substituted DC-81 and its dimers. AB - C2R-Fluoro substituted DC-81 and its dimers have been synthesized that exhibit significant DNA-binding ability, particularly the five carbon alkane spacer compound (6c) showed the helix melting temperature (DeltaTm) of 18.8 degrees C after incubation of 36 h at 37 degrees C. PMID- 17113292 TI - Discovery of [7-(2,6-dichlorophenyl)-5-methylbenzo [1,2,4]triazin-3-yl]-[4-(2 pyrrolidin-1-ylethoxy)phenyl]amine--a potent, orally active Src kinase inhibitor with anti-tumor activity in preclinical assays. AB - We describe the identification of [7-(2,6-dichlorophenyl)-5-methylbenzo [1,2,4]triazin-3-yl]-[4-(2-pyrrolidin-1-ylethoxy)phenyl]amine (3), a potent, orally active Src inhibitor with desirable PK properties, demonstrated activity in human tumor cell lines and in animal models of tumor growth. PMID- 17113293 TI - Endovascular embolization of ruptured cerebral aneurysms in patients older than 70 years. AB - The decision to manage ruptured cerebral aneurysms (RCAs) aggressively in elderly patients remains difficult because of inherent procedural risks and patient comorbidities. The purpose of this study was to report our experiences with and outcomes of endovascular embolization of RCAs in patients older than 70years. We studied 25 patients older than 70years with RCAs treated by endovascular embolization. There were 10 men and 15 women with a mean age of 77years. Subarachnoid haemorrhages of grades 1-3 were found in 24 patients, and one patient had grade 4 subarachnoid haemorrhage. Twenty-two RCAs were located in the anterior circulation, and three were in the posterior circulation. Seventeen RCAs were wide-necked aneurysms and five had to undergo stent-assisted coil embolization. Endovascular embolization was technically successful for all RCAs. Total angiographic obliteration of RCAs was achieved in eight (32%) cases, whereas the other 17 RCAs were subtotally or partially occluded. Procedure related death or severe disability occurred in two patients (8%). There was no rebleeding in any patient on follow-up. Endovascular embolization of RCAs has been proven to be both safe and effective in elderly patients, and should be performed in patients for whom surgical clipping would be difficult. PMID- 17113294 TI - Variations in the origin of the thalamoperforating arteries. AB - The anatomy of the thalamoperforating arteries located in the interpeduncular fossa must be well understood by surgeons to enable safe surgical treatment of basilar and posterior cerebral artery aneurysms. Therefore, we studied 30 posterior cerebral arteries obtained from 15 fresh adult cadaver brains. By filling the vertebral and internal carotid arteries of the brains with coloured latex, we found thalamoperforating arteries in 97% of the brains studied. The average number of arteries was two (range 0-5). Thalamoperforating arteries were classified into four different types according to their origin at the P1 segment: type I (bilateral multiple), 20%; type II (unilateral multiple, unilateral single), 33%; type III (bilateral single), 40%; type IV (one side multiple, the other side with no branches), 7%. In conclusion, it is important to bear in mind that these arteries can be the unilateral single type, and that they may be absent on the other side. Unilateral single arteries are very significant for surgical technique. PMID- 17113295 TI - Comparison of the histological and immunohistochemical features of the thymus in young- and elderly-onset myasthenia gravis without thymoma. AB - We performed histological and immunohistochemical analyses of the removed thymuses from 20 elderly (onset age > 60 years) and 23 young (onset age < 40 years) patients with myasthenia gravis (MG) who tested positive for serum anti acetylcholine receptor (AChR) antibodies, but who did not have associated thymoma. In the elderly group, nine (45%) patients had accumulations of lymphocytes, indicating an atrophied thymus with loss of the basic structure. The elderly MG patients with atrophied thymic tissues had higher titres of anti-AChR antibody (59.6+/-81.0 nmol/L) than those with adipose infiltration of the thymus alone (20.1+/-20.9 nmol/L). In immunohistochemical studies using image analysis, both young patients and elderly patients with atrophied thymic tissues were found to have significantly higher levels of CD20 than age-matched controls (p < 0.005). Atrophied thymic tissues, often seen immunohistochemically in young MG patients, may also be found in elderly patients, particularly in those with high titres of the anti-AChR antibody, even though adipose infiltration is marked in these patients. PMID- 17113296 TI - Biomechanical evaluation of different fixation plates in medial opening upper tibial osteotomy. AB - In this biomechanical study, 25 in vitro calf tibial models were used in order to compare the stability of the plates under axial compression loading. A 10-mm medial opening gap was stabilized in each of the five calf tibial models either with four or two-holed rectangular shaped plates with wedges, with four-holed reversed L-shaped plates with wedges, with the combination of these two types of plates, or with six-holed anatomical T-plates. The compression behavior of the model was tested by using a universal mechanical testing system. The specimens fixed with the combination of plates with the four-holed reversed L-shaped and with two-holed rectangular shaped; or with six-holed anatomical T-plates, showed significantly better stability than those of others. Four different kinds of failure (slippage of wedge, lateral cortex fracture, damage and/or loosening of screws, and bending of plates) were observed on the models. When the average value of force loading on the plates that were designed by the first author was considered, the plates were stable and the average force values at these points were higher than the loading force on a knee during the normal paced walking or running conditions. PMID- 17113297 TI - Haemorrhagic synovial cyst of the posterior cruciate ligament: a case report. AB - Cystic lesions arising in relation to the cruciate ligaments of the knee joint may become symptomatic and they can cause restriction of joint movement. We report here on a case of haemorrhagic synovial cyst arising from the posterior cruciate ligament with extension into the posterior compartment of the knee joint. We treated this lesion using the arthroscopic posterior-posterior triangulation technique. The histopathological findings of the synovial cyst were also verified. We would like to suggest that the posterior cruciate ligament synovial cyst should be considered in the differential diagnosis of cystic lesions that arise from the posterior compartment of the knee joint. PMID- 17113298 TI - Synthesis and biological evaluation of gamma-aminophosphonates as potent, subtype selective sphingosine 1-phosphate receptor agonists and antagonists. AB - The synthesis of N-arylamide phosphonates and related arylether and arylamine analogues provided potent, subtype-selective agonists and antagonists of the five known sphingosine 1-phosphate (S1P) receptors (S1P(1-5)). To this end, the syntheses of phosphoserine mimetics-selectively protected and optically active phosphonoserines-are described. In vitro binding assays showed that the implementation of phosphonates as phosphate mimetics provided compounds with similar receptor binding affinities as compared to their phosphate precursors. meta-substituted arylamide phosphonates were discovered to be antagonists of the S1P(1) and S1P(3) receptors. When administered to mice, an antagonist blocked the lymphopenia evoked by a S1P receptor agonist and caused capillary leakage in both lung and kidney. PMID- 17113299 TI - Molecular modeling of binding between amidinobenzisothiazoles, with antidegenerative activity on cartilage, and matrix metalloproteinase-3. AB - The aim of the work was to investigate the mechanism of binding between human metalloproteinase-3 (MMP-3) and new compounds belonging to the benzisothiazolylamidines class. In vitro tests suggest that these molecules, endowed with antinflammatory and cartilage antidegenerative activity, could act as ligands toward MMP-3. In lack of experimental structural informations, we performed molecular docking simulations to probe the interactions of benzisothiazolylamidines with matrix metalloproteinase-3, using the docking package GOLD and the software HINT as a post-process scoring function. Both GOLD and HINT predicted a binding mode for the compounds under analysis within the hydrophobic S1' pocket of MMP-3, without interaction with the catalytic Zn(2+) ion. The scores assigned by the programs to the interaction between the tested benzisothiazolylamidines and human MMP-3 were consistent with a potential direct enzyme inhibitory activity. The highest affinity was predicted for the N (benzo[d]isothiazol-3-yl)-4-chlorobenzamidine (2), emerged as the most active derivative also in the in vitro tests. PMID- 17113300 TI - Physicochemical and biological study of selected hydrophobic polyethylenimine based polycationic liposomes and their complexes with DNA. AB - Non-viral gene therapy is based on the development of efficient and safe gene carrier systems able to transfer DNA into cells. Polyethylenimine (PEI), the most promising non-viral vector, with its high cationic-charge-density potential is able (1) to compact DNA in complexes (polyplexes) smaller than those formed by liposomes (lipoplexes) and (2) to destabilize the endosomal membrane by a 'proton sponge' effect. Several PEI's hydrophobic modifications were reported in the last several years but in some cases a reduced transfection efficiency was observed. The mechanism underlying this phenomenon is not well understood so far. In order to extensively investigate these mechanisms, we reported a physicochemical and biological study of selected hydrophobic PEI's derivatives grafted with chains of different length and percentages of substitution able to form vesicles (polycationic liposomes) and to bind DNA. Their properties were studied by means of dynamic light scattering, freeze-fracture microscopy, potentiometric titrations, gel retardation assays, polyanion exchange reactions, toxicity assays, in vitro transfections, and fluorescence microscopy. Our results indicate that even if polyplexes are able to pass through the cellular membrane, the stability of PEI's hydrophobic polyplexes likely explain their different transfection efficiency in vitro. PMID- 17113301 TI - Lead optimization of [(S)-gamma-(arylamino)prolyl]thiazolidine focused on gamma substituent: Indoline compounds as potent DPP-IV inhibitors. AB - Dipeptidyl peptidase IV (DPP-IV) inhibitors are looked to as a potential new antidiabetic agent class. A series of [(S)-gamma-(arylamino)prolyl]thiazolidine compounds in which the electrophilic nitrile is removed are chemically stable DPP IV inhibitors. To discover a structure for the gamma-substituent of the proline moiety more suitable for interacting with the S(2) pocket of DPP-IV, optimization focused on the gamma-substituent was carried out. The indoline compound 22e showed a DPP-IV-inhibitory activity 100-fold more potent than that of the prolylthiazolidine 10 and comparable to that of NVP-DPP728. It also displayed improved inhibitory selectivity for DPP-IV over DPP8 and DPP9 compared to compound 10. Indoline compounds such as 22e have a rigid conformation with double restriction of the aromatic moiety by proline and indoline structures to promote interaction with the binding site in the S(2) pocket of DPP-IV. The double restriction effect provides a potent inhibitory activity which compensates for the decrease in activity caused by removing the electrophilic nitrile. PMID- 17113302 TI - High affinity Grb2-SH3 domain ligand incorporating Cbeta-substituted prolines in a Sos-derived decapeptide. AB - Peptide ligands that disrupt MAPK pathways are of great interest for a better understanding of these signalling cascades and represent therefore an attractive target to control cell degenerative processes. In that context, selective disruption of the upstream Grb2/Sos complex in the Ras/MAPK cascade has focused extensive work. The Sos PPII decapeptide, which interacts with the Grb2-SH3 domains, has been modified in various positions and the best inhibitors designed so far are either dimeric ligands or peptoid analogues of the VPPPVPPRRR sequence. We report the synthesis of new Grb2 ligands in which the key Val5 residue has been replaced by a cis C(beta)-substituted proline. Both fluorescence and ITC assays have been employed to measure the affinity of these substituted peptides for a recombinant Grb2 protein. Whereas proline in position 5 completely abolished the binding potency, a cis C(beta)-methyl-L-proline restored the affinity. Other cis C(beta)-proline substituents led to a complete loss of binding potency. Combining the best modifications: a cis C(beta)-methylproline 5, N-acetylation, C-carboxamide and dimerization yielded a 560-fold affinity enhancement compared to the wild-type VPPPVPPRRR sequence. This study shows that C(beta)-substituted prolines may constitute a new alternative for PPII ligands, combining entropy and enthalpy beneficial effects. PMID- 17113303 TI - Theoretical data of external Bremsstrahlung radiation cross-section of bone. AB - Theoretical data of external Bremsstrahlung (EB) radiation cross section of bone is estimated using tabulated results of EB cross section given for various elements at various photon and electron energies. This data may be useful in the analysis of Bremsstrahlung imaging which is the technique applied in medical therapy. PMID- 17113304 TI - Aortic valve surgery: time to be open-minded and to rethink. PMID- 17113305 TI - A new treatment option for pulmonary valvar insufficiency: first experiences with implantation of a self-expanding stented valve without use of cardiopulmonary bypass. AB - OBJECTIVE: Pulmonary regurgitation is the predominant problem in the long-term follow-up of tetralogy of Fallot (TOF) patients after primary repair. Apart from standard homograft implantation, a percutaneous valve delivery approach has been described recently. A right ventricular outflow tract (RVOT) diameter of greater than 22mm, however, precludes percutaneous valve delivery. We describe a novel technique with a transventricular implantation of a stented bio-prosthesis without cardiopulmonary bypass that allows for implantation of prosthesis with diameters greater than 22mm. METHODS: All patients (9-27 years of age) had undergone total correction of TOF at a mean age of 4.2+/-4.0 years. The RVOT was enlarged at that time with a transannular patch in all but one patient. All patients presented with severe pulmonary regurgitation without any significant RVOT obstruction. Mean MRI pulmonary regurgitation was 53+/-8%. The mean magnetic resonance imaging (MRI) right ventricular end diastolic volume index (RVEDVI) was 143+/-23ml/m(2), with a mean MRI right ventricular ejection fraction (RVEF) of 46+/-9%. In another two patients indication for treatment was based on reduced exercise capacity with patients being in NYHA Class III. After repeat sternotomy, a porcine valve mounted inside a self-expandable stent, covered with No-React treated porcine pericardium (Shelhigh, Model NR-4000MIS), was introduced just beneath the RVOT without use of cardiopulmonary bypass. External sutures were placed at the proximal and distal site of the valve to ensure fixation. RESULTS: The implantations were uneventful, with the patients hemodynamically stable throughout the procedure. One patient with severely dilated RVOT (up to 31mm) exhibited paravalvular leakage and the valve was replaced by a homograft after 2 days. At 6-12 month follow-up the remaining five patients exhibited no more than mild pulmonary regurgitation. The mean MRI RVEDVI was 94+/-18ml/m(2), with a mean MRI RVEF of 58+/-27%. CONCLUSIONS: Cardiopulmonary bypass for repeat RVOT interventions can be avoided in selected patients with this newly available device. In combination with a wide range of prosthesis sizes it offers yet another important treatment option. PMID- 17113306 TI - Glycyrrhizin inhibits the manifestations of anti-inflammatory responses that appear in association with systemic inflammatory response syndrome (SIRS)-like reactions. AB - In association with the systemic inflammatory response syndrome (SIRS), anti inflammatory response syndrome is commonly manifested in patients with trauma, burn injury, and after major surgery. These patients are increasingly susceptible to infection with various pathogens due to the excessive release of anti inflammatory cytokines from anti-inflammatory effector cells. Recently, CC chemokine ligand 2 (CCL2) found in the sera of mice with pancreatitis was identified as an active molecule for SIRS-associated anti-inflammatory response manifestation. Also, the inhibitory activity of glycyrrhizin (GL) on CCL2 production was reported. Therefore, the effect of GL on SIRS-associated anti inflammatory response manifestation was investigated in a murine SIRS model. Without any stimulation, splenic T cells from mice 5 days after SIRS induction produced cytokines associated with anti-inflammatory response manifestation. However, these cytokines were not produced by splenic T cells from SIRS mice previously treated with GL. In dual-chamber transwells, IL-4-producing cells were generated from normal T cells cultured with peripheral blood polymorphonuclear neutrophils (PMN) from SIRS mice. However, IL-4-producing cells were not generated from normal T cells in transwell cultures performed with PMN from GL treated SIRS mice. CCL2 was produced by PMN from SIRS mice, while this chemokine was not demonstrated in cultures of PMN from SIRS mice treated with GL. These results indicate that GL has the capacity to suppress SIRS-associated anti inflammatory response manifestation through the inhibition of CCL2 production by PMN. PMID- 17113307 TI - Exploring the energy landscape of protein folding using replica-exchange and conventional molecular dynamics simulations. AB - Two independent replica-exchange molecular dynamics (REMD) simulations with an explicit water model were performed of the Trp-cage mini-protein. In the first REMD simulation, the replicas started from the native conformation, while in the second they started from a nonnative conformation. Initially, the first simulation yielded results qualitatively similar to those of two previously published REMD simulations: the protein appeared to be over-stabilized, with the predicted melting temperature 50-150K higher than the experimental value of 315K. However, as the first REMD simulation progressed, the protein unfolded at all temperatures. In our second REMD simulation, which starts from a nonnative conformation, there was no evidence of significant folding. Transitions from the unfolded to the folded state did not occur on the timescale of these simulations, despite the expected improvement in sampling of REMD over conventional molecular dynamics (MD) simulations. The combined 1.42 micros of simulation time was insufficient for REMD simulations with different starting structures to converge. Conventional MD simulations at a range of temperatures were also performed. In contrast to REMD, the conventional MD simulations provide an estimate of Tm in good agreement with experiment. Furthermore, the conventional MD is a fraction of the cost of REMD and continuous, realistic pathways of the unfolding process at atomic resolution are obtained. PMID- 17113308 TI - Oral and anal immunisation with alloantigen induces active cell-mediated cytotoxic responses in carp. PMID- 17113309 TI - Intentions and expectations in temporal binding. AB - Recently, it has been shown that the perceived times of voluntary movements and their effects are perceived as shifted towards each other. This temporal binding phenomenon was explained by an integrated representation of movement and effect, facilitating operant learning and the experience of intentionality. Here, we investigated whether temporal binding depends on explicit intentional attributions. In Experiment 1, participants intended to either produce or avoid producing an effect (a tone) by the timing of their movements, with the ratio of success being fixed at 2:1. In Experiments 2 and 3, the influence of the action effect contingency ratio on temporal binding was controlled for by removing the intentional attribution of the effect. The results indicate that temporal binding is a general associative mechanism that facilitates the learning of movement effect contingencies. Beyond that, temporal binding is sensitive to explicit intentional attributions, which selectively enhance the link between an intentional movement and the effect a moving agent intends to produce. PMID- 17113310 TI - Metabolic brain networks associated with cognitive function in Parkinson's disease. AB - The motor manifestations of Parkinson's disease (PD) have been linked to an abnormal spatial covariance pattern involving basal ganglia thalamocortical pathways. By contrast, little is known about the functional networks that underlie cognitive dysfunction in this disorder. To identify such patterns, we studied 15 non-demented PD patients using FDG PET and a voxel-based network modeling approach. We detected a significant covariance pattern that correlated (p<0.01) with performance on tests of memory and executive functioning. This PD related cognitive pattern (PDCP) was characterized by metabolic reductions in frontal and parietal association areas and relative increases in the cerebellar vermis and dentate nuclei. To validate this pattern, we analyzed data from 32 subsequent PD patients of similar age, disease duration and severity. Prospective measurements of PDCP activity predicted memory performance (p<0.005), visuospatial function (p<0.01), and perceptual motor speed (p<0.005) in this validation sample. PDCP scores additionally exhibited an excellent degree of test retest reliability (intraclass correlation coefficient, ICC=0.89) in patients undergoing repeat FDG PET at an 8-week interval. Unlike the PD-related motor pattern, PDCP expression was not significantly altered by antiparkinsonian treatment with either intravenous levodopa or deep brain stimulation (DBS). These findings substantiate the PDCP as a reproducible imaging marker of cognitive function in PD. Because PDCP expression is not altered by routine antiparkinsonian treatment, this measure of network activity may prove useful in clinical trials targeting the progression of non-motor manifestations of this disorder. PMID- 17113311 TI - Measuring brain variability by extrapolating sparse tensor fields measured on sulcal lines. AB - Modeling and understanding the variability of brain structures is a fundamental problem in neurosciences. Improved mathematical representations of structural brain variation are needed to help detect and understand genetic or disease related sources of abnormality, as well as to improve statistical power when integrating functional brain mapping data across subjects. In this paper, we develop a new mathematical model of normal brain variation based on a large set of cortical sulcal landmarks (72 per brain) delineated in each of 98 healthy human subjects scanned with 3D MRI (age: 51.8+/-6.2 years). We propose an original method to compute an average representation of the sulcal curves, which constitutes the mean anatomy. After affine alignment of the individual data across subjects, the second order moment distribution of the sulcal position is modeled as a sparse field of covariance tensors (symmetric, positive definite matrices). To extrapolate this information to the full brain, one has to overcome the limitations of the standard Euclidean matrix calculus. We propose an affine invariant Riemannian framework to perform computations with tensors. In particular, we generalize radial basis function (RBF) interpolation and harmonic diffusion partial differential equations (PDEs) to tensor fields. As a result, we obtain a dense 3D variability map that agrees well with prior results on smaller subject samples. Moreover, "leave one (sulcus) out" tests show that our model is globally able to recover the missing information on brain variation when there is a consistent neighboring pattern of variability. Finally, we propose an innovative method to analyze the asymmetry of brain variability. As expected, the greatest asymmetries are found in regions that includes the primary language areas. Interestingly, any such asymmetries in anatomical variance, if it remains after anatomical normalization, could explain why there may be greater power to detect group activation in one hemisphere versus the other in fMRI studies. PMID- 17113312 TI - Strategies for attenuation compensation in neurological PET studies. AB - Molecular brain imaging using positron emission tomography (PET) has evolved into a vigorous academic field and is progressively gaining importance in the clinical arena. Significant progress has been made in the design of high-resolution three dimensional (3-D) PET units dedicated to brain research and the development of quantitative imaging protocols incorporating accurate image correction techniques and sophisticated image reconstruction algorithms. However, emerging clinical and research applications of molecular brain imaging demand even greater levels of accuracy and precision and therefore impose more constraints with respect to the quantitative capability of PET. It has long been recognized that photon attenuation in tissues is the most important physical factor degrading PET image quality and quantitative accuracy. Quantitative PET image reconstruction requires an accurate attenuation map of the object under study for the purpose of attenuation compensation. Several methods have been devised to correct for photon attenuation in neurological PET studies. Significant attention has been devoted to optimizing computational performance and to balancing conflicting requirements. Approximate methods suitable for clinical routine applications and more complicated approaches for research applications, where there is greater emphasis on accurate quantitative measurements, have been proposed. The number of scientific contributions related to this subject has been increasing steadily, which motivated the writing of this review as a snapshot of the dynamically changing field of attenuation correction in cerebral 3D PET. This paper presents the physical and methodological basis of photon attenuation and summarizes state of the art developments in algorithms used to derive the attenuation map aiming at accurate attenuation compensation of brain PET data. Future prospects, research trends and challenges are identified and directions for future research are discussed. PMID- 17113313 TI - Analysis of oxygen metabolism implies a neural origin for the negative BOLD response in human visual cortex. AB - The sustained negative blood oxygenation level-dependent (BOLD) response in functional MRI is observed universally, but its interpretation is controversial. The origin of the negative response is of fundamental importance because it could provide a measurement of neural deactivation. However, a substantial component of the negative response may be due to a non-neural hemodynamic artifact. To distinguish these possibilities, we have measured evoked BOLD, cerebral blood flow (CBF), and oxygen metabolism responses to a fixed visual stimulus from two different baseline conditions. One is a normal resting baseline, and the other is a lower baseline induced by a sustained negative response. For both baseline conditions, CBF and oxygen metabolism responses reach the same peak amplitude. Consequently, evoked responses from the negative baseline are larger than those from the resting baseline. The larger metabolic response from negative baseline presumably reflects a greater neural response that is required to reach the same peak amplitude as that from resting baseline. Furthermore, the ratio of CBF to oxygen metabolism remains approximately the same from both baseline states (approximately 2:1). This tight coupling between hemodynamic and metabolic components implies that the magnitude of any hemodynamic artifact is inconsequential. We conclude that the negative response is a functionally significant index of neural deactivation in early visual cortex. PMID- 17113314 TI - Phylogeographic incongruence of codistributed amphibian species based on small differences in geographic distribution. AB - Codistributed species may display either congruent phylogeographic patterns, indicating similar responses to a series of shared climatic and geologic events, or discordant patterns, indicating independent responses. This study compares the phylogeographic patterns of two similarly distributed salamander species within the Pacific Northwest of the United States: Cope's giant salamander (Dicamptodon copei) and Van Dyke's salamander (Plethodon vandykei). Previous studies of P. vandykei support two reciprocally monophyletic lineages corresponding to coastal populations, located from the Olympic Mountains to the mouth of the Columbia River, and inland populations within the Cascade Mountains. We hypothesized that D. copei would have a congruent phylogeographic pattern to P. vandykei due to similarity in distribution and dependence upon similar stream and stream-side habitats. We test this hypothesis by estimating the phylogeny of D. copei using approximately 1800bp of mitochondrial DNA and comparing it to that of P. vandykei. Sympatric populations of D. copei and of P. vandykei display an identical phylogeographic pattern, suggesting similar responses within their shared distribution. Populations of D. copei occurring outside the range of P. vandykei displayed high levels of genetic divergence from those sympatric to P. vandykei. Overall, phylogeographic patterns between the two species were ultimately incongruent due to the high divergence of these allopatric populations. These results provide an example of codistributed species displaying overall incongruent phylogeographic patterns while simultaneously displaying congruent patterns within portions of their shared geographic distribution. This pattern demonstrates that a simple dichotomy of congruent and incongruent phylogeographic patterns of codistributed species may be too simplistic and that more complex intermediate patterns can result even from minor differences in species' ranges. PMID- 17113315 TI - Combined mitochondrial and nuclear sequences support the monophyly of forcipulatacean sea stars. AB - Previous molecular phylogenetic analyses of forcipulatacean sea stars (Echinodermata: Asteroidea) have reconstructed a non-monophyletic order Forcipulatida, provided that two or more forcipulate families are included. This result could mean that one or more assumptions of the reconstruction method was violated, or else the traditional classification could be erroneous. The present molecular phylogenetic analysis included 12 non-forcipulatacean and 39 forcipulatacean sea stars, with multiple representatives of all but one of the forcipulate families and/or subfamilies. Bayesian analysis of approximately 4.2kb of sequence data representing seven partitions (nuclear 18S rRNA and 28S rRNA, mitochondrial 12S rRNA, 16S rRNA, 5 tRNAs and cytochrome oxidase I with first and second codon positions analyzed separately from third codon positions) recovered a consensus tree with three well-supported clades (78%-100% bootstrap support) that corresponded at least approximately to traditional taxonomic ranks: the superorder Forcipulatacea (Forcipulatida + Brisingida) + Pteraster, the Brisingida/Brisingidae and Asteriidae + Rathbunaster + Pycnopodia. When a molecular clock was enforced, the partitioned Bayesian analysis recovered the traditional Forcipulatacea. Five of six genera represented by two or more species were monophyletic with 100% bootstrap support. Most of the traditional subfamilial and familial groupings within the Forcipulatida were either unresolved or non-monophyletic. The separate partitions differed considerably in estimates of model parameters, mainly between nuclear sequences (with high GC content, low rates of sequence substitution and high transition/transversion rate ratios) and mitochondrial sequences. PMID- 17113316 TI - How and when did Old World ratsnakes disperse into the New World? AB - To examine Holarctic snake dispersal, we inferred a phylogenetic tree from four mtDNA genes and one scnDNA gene for most species of the Old World (OW) and New World (NW) colubrid group known as ratsnakes. Ancestral area distributions are estimated for various clades using divergence-vicariance analysis and maximum likelihood on trees produced using Bayesian inference. Dates of divergence for the same clades are estimated using penalized likelihood with statistically crosschecked calibration references obtained from the Miocene fossil record. With ancestral areas and associated dates estimated, various hypotheses concerning the age and environment associated with the origin of ratsnakes and the dispersal of NW taxa from OW ancestors were tested. Results suggest that the ratsnakes originated in tropical Asia in the late Eocene and subsequently dispersed to the Western and Eastern Palearctic by the early Oligocene. These analyses also suggest that the monophyletic NW ratsnakes (the Lampropeltini) diverged from OW ratsnakes and dispersed through Beringia in the late Oligocene/early Miocene when this land bridge was mostly composed of deciduous and coniferous forests. PMID- 17113317 TI - Shoulder arthroplasty in cases with avascular necrosis of the humeral head. AB - Avascular necrosis (AVN) is a relatively uncommon cause of glenohumeral arthritis. Previous retrospective reviews of shoulder arthroplasty for AVN have shown very good results in small numbers of patients. This study prospectively evaluated a consecutive series of 21 shoulders in 19 patients treated with the same modular prosthesis at a single institution. Of the shoulders, 8 developed AVN after a proximal humeral fracture, 1 was associated with a massive rotator cuff tear, 10 developed after corticosteroid therapy, and 2 were idiopathic. The patients, 14 women and 5 men, with a mean age of 54 years, were followed up for a mean of 4.7 years (range, 2 to 8 years). Hemiarthroplasty was performed in 15 shoulders, whereas 6 required total shoulder arthroplasty. Assessment included visual analog scales, the Simple Shoulder Test, and the American Shoulder and Elbow Surgeons Shoulder Score Index (SSI). Physical examination was performed, and radiographs were obtained. The visual analog scale scores for pain and function improved significantly after surgery (P < .01). The mean SSI score rose from 36 to 81 (where 0 indicates the worst outcome and 100 indicates the best outcome). The mean number of positive responses on the Simple Shoulder Test increased from 3 to 10. Mean active elevation increased from 88 degrees to 123 degrees . External rotation improved from 7 degrees to 34 degrees , and internal rotation improved from L4 to T12. No difference in outcome was noted based on age or sex. Patients with steroid-related or idiopathic AVN had greater pain and functional impairment preoperatively (mean SSI score, 26) than those with prior fracture or rotator cuff tear (mean SSI score, 49) (P < .01). Postoperatively, however, these groups fared equally well (mean SSI score, 78 vs 82). Shoulder arthroplasty for AVN produces good pain relief and function, although a concurrent series with osteoarthritis yielded better results. Forward elevation did not approach normal as had been previously reported. PMID- 17113318 TI - An in vivo comparison of the modified Mason-Allen suture technique versus an inclined horizontal mattress suture technique with regard to tendon-to-bone healing: a biomechanical and histologic study in sheep. AB - The purpose of this study is to examine long-term tendon-to-bone healing, by use of a sheep animal model, after rotator cuff repairs performed with 2 different suture techniques: an inclined horizontal mattress suture pattern placed with special arthroscopic instrumentation (HMS) and the modified Mason-Allen pattern (MMA). After a pre hoc power analysis, 18 skeletally mature sheep were randomly assigned to either the HMS or MMA repair technique, with contralateral limbs used for the control group. At 26 weeks, the animals were euthanized. Six sheep from each group underwent biomechanical testing. Load-to-failure and stiffness results indicated no statistically significant difference between the 2 groups. Avulsion of the tuberosity was the primary mode of failure for both groups. In the remaining 6 sheep, histologic evaluation demonstrated that, regardless of treatment, the tendon appeared completely healed in the bony trough. Because the long-term biomechanical and histologic properties of healed tendons repaired with an HMA technique are equal to those obtained with an MMA technique, the inclined horizontal mattress suture may be appropriate for arthroscopic rotator cuff repair. Short-term studies are necessary to determine whether these findings are true early after tendon repair, when failure may be most common. PMID- 17113319 TI - Arthroscopic repair of partial-thickness tears of the rotator cuff. AB - This study prospectively evaluated the clinical outcome of 41 patients who underwent arthroscopic repair of a significant (>50% thickness) partial-thickness supraspinatus tear. Tears were converted to full-thickness lesions and repaired with suture anchors and simple sutures. Of the tears, 80% (33/41) involved the articular surface. The mean age of the patients was 49 years (range, 23-70 years), and the mean follow-up was 38 months (range, 24-50 months). All patients had improvements in range of motion and strength. Postoperative isometric strength measurements revealed no significant difference between the operative and asymptomatic shoulder. Significant improvements were demonstrated for American Shoulder and Elbow Surgeons scores (from 42 to 93 points, P < .001), pain relief (from 6.5 to 0.8 points, P < .001), and satisfaction (from 3.0 to 9.2 points, P < .001). Of the 41 patients, 40 (98%) were satisfied with their outcome. This study documented the successful clinical outcome of arthroscopic repair of significant partial-thickness rotator cuff tears. PMID- 17113320 TI - The effect of cartilage-derived morphogenetic protein 2 on initial healing of a rotator cuff defect in a rat model. AB - This animal study evaluated the healing of supraspinatus tendon tears by use of a cartilage-derived morphogenetic protein 2 growth factor (CDMP-2) delivered to the repair. Forty-eight rats had bilateral, surgically created complete tears repaired by sutures with the growth factor introduced on one side. They were killed at 2, 3, 4, and 6 weeks, and the strength of the repairs was determined and histologic analysis performed. At 4 and 6 weeks, the CDMP-2-treated repairs were significantly stronger than the untreated repairs and histologic analysis showed more organized healing. The use of growth factors introduced at the time of rotator cuff repair might promote more rapid healing and subsequent, rapid patient rehabilitation. PMID- 17113321 TI - Early complications of operatively treated proximal humeral fractures. AB - Minimal information exists regarding early complications after operatively treated proximal humeral fractures. Of the 82 shoulders that had osteosynthesis, 42 had a (nonmedical) complication, with 21 requiring further surgery. Of 42 shoulders with complications, 12 were related to incomplete reduction, 16 had loss of anatomic fracture fixation, 9 had delayed healing, 3 had an infection, 1 had rotator cuff failure, and 1 had loose bodies. Fixed-angle plates had lower rates of initial malpositioning and resultant malunion. Of the 22 shoulders requiring hemiarthroplasty, 14 had an early complication. Of these, 7 had complications relating to implant insertion or tuberosity malreduction at the index operation and 7 had problems with tuberosity healing. The rate of complications after operative treatment of proximal humeral fractures is high. All efforts at fracture fragment fixation with osteosynthesis and hemiarthroplasty should be directed at obtaining anatomic fracture fixation that resists displacement. PMID- 17113322 TI - Surgical treatment of varus malunion of the proximal humerus with valgus osteotomy. AB - Among the complications of 2-part fractures of the upper end of the humerus, little attention has been paid to the treatment of the varus malunion. However, this deformity frequently causes pain and disability of the shoulder. To improve this condition, we treated this malunion with a valgus wedge osteotomy. From August 1995 to January 1999, 5 patients with this deformity, all complaining about pain or unsatisfactory function of the shoulder (University of California, Los Angeles score <13), underwent osteotomy and internal fixation with a plate and screws. The mean age was 53 years (range, 25-73 years), there were 4 male patients, and the left side was involved in 4 cases. The mean follow-up was 34 months (range, 22-63 months). Union occurred in all cases by 6 weeks. Three of the results were excellent, and two were good (University of California, Los Angeles score >30). All patients were satisfied with the treatment. PMID- 17113323 TI - The influence of the acromial coverage index in rotator cuff tears. AB - Several intrinsic and extrinsic factors have been advocated in the pathogenesis of rotator cuff tears, but it is still unclear whether the origin of the tear is related to tendon degeneration itself or induced by several morphologic changes. The purpose of this study is to determine the relationship between the acromial coverage of the humeral head and the presence of a cuff tear. We evaluated 148 shoulders, including 45 that underwent surgical rotator cuff repair (group I), 26 with documented rotator cuff tears treated conservatively (group II), and 77 with no cuff pathology as a control group (group III). The mean acromial coverage index was 0.68 in group I, 0.72 in group II, and 0.59 in group III, giving a highly significant difference (P < .0001) between the control group and both cuff tear groups. Patients with a cuff tear have a significantly higher acromial coverage index than the control group. PMID- 17113324 TI - Three-dimensional rotation of the scapula during functional movements: an in vivo study in healthy volunteers. AB - The goal of this study was to measure 3-dimensional shoulder motion by use of a direct invasive technique during 4 different arm movements in healthy volunteers. Eight subjects with healthy shoulders were recruited. Optoelectronic marker carriers (ie, infrared light-emitting diodes) were mounted on bone pins, which were inserted into the lateral scapular spine. Subjects performed 4 different arm movements while the motion was being recorded by a precision optoelectronic camera. Joint angles were calculated in 3 dimensions. Intraclass correlation coefficients and root-mean-square differences were calculated as measures of reliability. During abduction, the scapula tipped posteriorly (44 degrees +/- 11 degrees), rotated upward (49 degrees +/- 7 degrees), and rotated externally (27 degrees +/- 11 degrees). For reaching, the scapula consistently rotated upward (17 degrees +/- 3 degrees) and rotated internally (18 degrees +/- 6 degrees) whereas tipping was generally less than 10 degrees (5 degrees +/- 2 degrees). Overall, the range of scapular movement for the hand behind the back was small and variable, with most rotations not exceeding 15 degrees. For horizontal adduction, the scapula tipped anteriorly (8 degrees +/- 3 degrees), rotated upward (5 degrees +/- 2 degrees), and rotated internally (27 degrees +/- 6 degrees). These scapular rotations provide normative data that will be useful for diagnosing scapular dysfunction. PMID- 17113325 TI - The Genetics of Generalized Osteoarthritis (GOGO) study: study design and evaluation of osteoarthritis phenotypes. AB - PURPOSE: The primary goal of the Genetics of Generalized Osteoarthritis (GOGO) study is to identify chromosomal regions associated with increased susceptibility to generalized osteoarthritis (OA). Here we describe the study design and phenotype of the 2728 participants from the 1145 families recruited for this study. METHODS: GOGO is an investigator-initiated collaboration involving seven clinical academic sites and sponsored by GlaxoSmithKline. Family ascertainment was carried out between 1999 and 2002. A qualifying family required self-reported Caucasian ethnicity and at least two affected siblings with clinical hand OA. We hypothesized that this clinical phenotype would facilitate identification of participants with multijoint radiographic OA (rOA) in and beyond the hand. The "gold standard" case definition, however, was based on rOA (Kellgren-Lawrence grade > or =2) involving > or =3 hand joints distributed bilaterally and including at least one distal interphalangeal joint, with two of the three involved joints within a joint group (distal interphalangeal, proximal interphalangeal, or carpometacarpal). Radiographs of hips, knees and spine were also obtained. Additional siblings and living parents from qualifying families, both affected and unaffected, were invited to participate. RESULTS: A total of 2706 participants had complete clinical and radiological examination data. Of these, 2569 participants met clinical examination criteria for affected status; while 1963 (73%) participants met the prespecified radiographic criteria for affected status. This corresponded to a total of 707 families with at least two affected siblings that met the hand rOA criteria. Of those individuals with rOA of the hand, the frequency of rOA at other sites was highest for the knee (51%) and spine (54%), and less common for the hip (25%). Concordance rates among hand affected siblings were greatest for spine (36%) followed by knee (31%) and hip (9%); a total of 53% of the affected sib pairs were concordant for specific patterns of generalized rOA involving the hand and large joints (knees, hips or spine). CONCLUSIONS: GOGO represents a large multicenter collection of families with multiple joint OA that have been characterized both clinically and radiographically. The GOGO study will employ a comprehensive strategy for genetic screening based upon both qualitative and quantitative radiographic trait analyses, circulating biomarkers in a quantitative trait-based analysis, fine mapping, and candidate gene analysis. This sample should provide sufficient power to detect linkage to OA associated genes. PMID- 17113326 TI - Isolation of an isoflavone-metabolizing, Clostridium-like bacterium, strain TM 40, from human faeces. AB - Recently, the biological effects of isoflavones have attracted much attention. Intestinal microbiota plays an important role in the metabolism and bioavailability of isoflavones. However, few reports have discussed intestinal bacteria that metabolize daidzein into dihydrodaidzein. In this study, we isolated the dihydrodaidzein-producing intestinal bacterium TM-40 from a healthy boy's faeces. The bacteria from faecal samples were incubated with daidzein. Among all tested bacteria, one strain (strain TM-40) produced dihydrodaidzein both from daidzein and daidzin. However, in our experimental conditions, strain TM-40 did not produce equol from daidzein. The 16S rRNA partial sequence of strain TM-40 (AB249652) exhibited a 93% similarity to that of Coprobacillus catenaformis (AB030218). This strain seems to be a new species. PMID- 17113327 TI - RNA targeting with peptide conjugates of oligonucleotides, siRNA and PNA. AB - Towards the development of oligonucleotide analogues and siRNA as drugs, one potential alternative to the use of liposomal transfection agents is the covalent conjugation of a cell-penetrating peptide (CPP), with the intention of imparting on the oligonucleotide or siRNA an enhanced ability to enter mammalian cells and reach the appropriate RNA target. We have developed robust methods for the chemical synthesis of disulfide-linked conjugates of oligonucleotide analogues, siRNA and peptide nucleic acids (PNA) with a range of cationic and other CPPs. In a HeLa cell assay with integrated plasmid reporters of Tat-dependent trans activation at the TAR RNA target in the cell nucleus, we were unable to obtain steric block inhibition of gene expression for conjugates of CPPs with a 12-mer oligonucleotide mixmer of 2'-O-methyl and locked nucleic acids units. By contrast, we were able to obtain some reductions in expression of P38alpha MAP kinase mRNA in HeLa cells using microM concentrations of Penetratin or Tat peptides conjugated to the 3'-end of the sense strand of siRNA. However, the most promising results to date have been with a 16-mer PNA conjugated to the CPP Transportan or a double CPP R(6)-Penetratin, where we have demonstrated Tat dependent trans-activation inhibition in HeLa cells. Results to date suggest the possibility of development of CPP-PNA conjugates as anti-HIV agents as well as other potential applications involving nuclear cell delivery, such as the redirection of splicing. PMID- 17113328 TI - Blockade of metabotropic glutamate receptor 5 activation inhibits mechanical hypersensitivity following abdominal surgery. AB - This study used the metabotropic glutamate 5 (mGlu5) receptor subtype-selective antagonist 2-methyl-6-(phenylethynyl)pyridine (MPEP) to characterise the contribution of mGlu5 receptor activity to pain and hypersensitivity in an animal model of post-surgical pain. Adult female Wistar rats (200-250g) were anaesthetised with isoflurane (2%) and underwent a midline laparotomy with gentle manipulation of the viscera, and the effects of pre- (30min) or post- (5h) operative treatment with MPEP (1, 3 or 10mgkg(-1); i.p.) or drug-vehicle on hindpaw withdrawal latency (in seconds) to thermal stimulation (Hargreave's Test) and response threshold (in grams) to mechanical stimulation (using a dynamic plantar aesthesiometer) were measured. Animals that underwent surgery displayed significant hypersensitivity to mechanical stimulation of the hindpaws. Hypersensitivity was maximum at 6h post-surgery (44.5+/-2.4% decrease; p<0.01 vs. anaesthesia only controls) and persisted for 48h. Surgery had no effect on thermal withdrawal latency. Both pre-operative and post-operative administration of 10mgkg(-1)MPEP blocked mechanical hypersensitivity induced by surgery (p<0.01 vs. vehicle treatment). MPEP had no effect on acute nociceptive thresholds in naive animals. These data suggest that activity at mGlu5 receptors contributes to development of pain and hypersensitivity following surgery. PMID- 17113329 TI - Long term methadone for chronic pain: a pilot study of pharmacokinetic aspects. AB - Methadone is used as an alternative opioid when first line opioids fail to provide adequate pain control. Highly variable morphine:methadone dose ratios make switching challenging and little is known about the pharmacokinetics of long lasting methadone treatment for pain. Twelve patients treated with morphine for chronic non-malignant pain were switched to methadone. Seven of these patients continued with methadone throughout the nine months study period and only minor dose adjustments were performed. Serum concentrations of morphine, methadone and their metabolites were measured at baseline, day one and two, after dose titration and one week, five weeks, three months and nine months after the end of dose titration. Serum concentrations of methadone and its metabolite EDDP did not change significantly from the end of dose titration and during the nine months (repeated measures ANOVA: p=0.88 and p=0.06). Very low correlation between dose ratios and serum concentration ratios between morphine and methadone was observed. Large interindividual differences in serum concentrations and metabolism were observed. Our findings contradict that autoinduction of methadone metabolism takes place during long term treatment and supports that a 3-day opioid switch from morphine to methadone followed by a one week titration seems pharmacologically sound. PMID- 17113330 TI - Clinical assessment of chronic cough severity. AB - The assessment of cough severity solely through consultation with the patient is limited by its subjective nature and variability of physician judgement. The assessment of cough has been hampered by a paucity of objective tools, those available have been poorly validated, non-standardized and are impractical for clinical use. Recent developments have overcome some of these problems and a variety of tools can now be used to assess cough that include visual analogue scales (VAS), quality-of-life questionnaires, cough reflex sensitivity measurement and automated cough frequency monitors. These tools can be used to validate the presence of cough and assess response to therapy. They will also have an important role in clinical trials. PMID- 17113331 TI - [Present and future of the image guided radiotherapy (IGRT) and its applications in lung cancer treatment]. AB - These last years, the new irradiation techniques as the conformal 3D radiotherapy and the IMRT are strongly correlated with the technological developments in radiotherapy. The rigorous definition of the target volume and the organs at risk required by these irradiation techniques, imposed the development of various image guided patient positioning and target tracking techniques. The availability of these imaging systems inside the treatment room has lead to the exploration of performing real-time adaptive radiation therapy. In this paper we present the different image guided radiotherapy (IGRT) techniques and the adaptive radiotherapy (ART) approaches. IGRT developments are focused in the following areas: 1) biological imaging for better definition of tumor volume; 2) 4D imaging for modeling the intra-fraction organ motion; 3) on-board imaging system or imaging devices registered to the treatment machines for inter-fraction patient localization; and 4) treatment planning and delivery schemes incorporating the information derived from the new imaging techniques. As this paper is included in the "Cancer-Radiotherapie" special volume dedicated to the lung cancers, in the description of the different IGRT techniques we try to present the lung tumors applications when this is possible. PMID- 17113332 TI - Salmonella Typhimurium SPI-1 genes promote intestinal but not tonsillar colonization in pigs. AB - Salmonella Pathogenicity Island 1 (SPI-1) genes are indispensable for virulence of Salmonella Typhimurium in several animal species. The role of SPI-1 in the pathogenesis of Salmonella Typhimurium infections of pigs, however, is not well described. The interactions of a porcine Salmonella Typhimurium field strain and its isogenic mutants with disruptions in the SPI-1 genes hilA, sipA and sipB with porcine intestinal epithelial cells were characterized in vitro and in a ligated intestinal loop model in pigs. HilA and SipB were essential in the invasion of porcine intestinal epithelial cells in vitro. A sipA mutant was impaired for invasion using a polarized cell line, but fully invasive in a non-polarized cell line. All SPI-1 mutants induced a significant decrease in influx of neutrophils in the porcine intestinal loop model compared with the wild type strain. Pigs were orally inoculated with 10(8) colony forming units of both the wild type Salmonella Typhimurium strain and its isogenic sipB::kan mutant strain. The sipB mutant strain was significantly impaired to invade the intestinal, but not the tonsillar tissue, one day after inoculation and was unable to efficiently colonize the intestines and the GALT, but not the tonsils, 3 days after inoculation. This study shows that SPI-1 plays a crucial role in the invasion and colonization of the porcine gut and in the induction of influx of neutrophils towards the intestinal lumen, but not in the colonization of the tonsils. PMID- 17113333 TI - Analysis of HIV-1 sequences before and after co-infecting syphilis. AB - Increasing syphilis incidence among men who have sex with men (MSM) has been reported. The index case was a human immunodeficiency virus type 1 (HIV-1) positive MSM who presented coincidentally with the secondary syphilis and a rebound of plasma viral load after complete suppression of HIV-1 (below 50 copies/ml) for 13 months with potent antiretroviral therapy (PART), suggesting a possibility of HIV-1 superinfection. We analyzed HIV-1 sequences before and after syphilis in four HIV-1-positive patients including the index case to explore drug resistance mutations (DRMs) and a possibility of HIV-1 superinfection. There were patients who obtained DRMs around syphilis infection but no evidence of HIV-1 superinfection was obtained. Our results underline the importance of strict adherence to PART. PMID- 17113334 TI - Synthesis of 1,5-dinitroaryl-1,4-pentadien-3-ones under ultrasound irradiation. AB - 1,5-Dinitroaryl-1,4-pentadien-3-ones were synthesized with ultrasound irradiation in the presence of K(2)CO(3) as catalyst. The reaction mechanism and the factors influencing the product were also discussed. The present procedure is more convenient with shorter reaction time and higher yields compared with conventional methods. PMID- 17113335 TI - The correlation between rates of unemployment and suicides rates in Japan between 1984 and 2003. PMID- 17113336 TI - Inhibition of plasma membrane Ca2+-ATPase by heparin is modulated by potassium. AB - Heparin is related to several protein receptors that control Ca2+ homeostasis. Here, we studied the effects of heparin on the plasma membrane Ca2+-ATPase from erythrocytes. Both ATP hydrolysis and Ca2+ uptake were inhibited by heparin without modification of the steady-state level of phosphoenzyme formed by ATP. Calmodulin did neither modify the inhibition nor the binding of heparin. Inhibition by heparin was counteracted by K+ but not by Li+. This effect was extended to other sulfated polysaccharides with high number of sulfate residues. Hydrolysis of p-nitrophenylphosphate was equally inhibited by heparin. No evidence for enzyme uncoupling was observed: Ca2+ uptake and ATP hydrolysis remained tightly associated at any level of heparin, and heparin did not increase the passive Ca2+ efflux of inside-out vesicles. Vanadate blocked this efflux, indicating that the main point of Ca2+ escape from these vesicles was linked to the Ca2+ pump. It is discussed that sulfated polysaccharides may physiologically increase the steady-state level of Ca2+ in the cytosol by inhibiting the Ca2+ pumps in a K+ (and tissue) regulated way. It is suggested that heparin regulates the plasma membrane Ca2+-ATPase by binding to the E2 conformer. PMID- 17113337 TI - Insulin-like growth factors control cell migration in health and disease. AB - Insulin-like growth factors I and II (IGF-I and IGF-II) have an ancient origin and play essential roles in fundamental biological processes. Although IGFs are principally known for their roles in regulating cell growth and survival, their ability to influence cell motility is just as significant. In the past 20 years, research has provided indisputable evidence for the regulatory role of IGFs in the migration of various cell types. Cell migration is crucial for reproduction, development, and tissue regeneration; IGFs play an important role in coordinating these processes. Moreover, studies continue to uncover the IGFs' role in stimulating cancer cell migration, invasion and metastasis. This review surveys current knowledge on the cell migration-modulating properties of IGFs and the biochemical pathways by which these peptides regulate cell movement in both physiological and pathological conditions. PMID- 17113338 TI - The regulation and role of osteopontin in malignant transformation and cancer. AB - Osteopontin (OPN) is a predominantly secreted extracellular matrix glycophosphoprotein which binds to alpha v-containing integrins and has an important role in malignant cell attachment and invasion. High OPN expression in the primary tumor is associated with early metastasis and poor outcome in human breast and other cancers. Forced OPN overexpression in benign cells may induce neoplastic-like cell behaviour including increased attachment and invasion in vitro as well as the ability to metastasize in vivo. Conversely, OPN inhibition by antisense cDNA impedes cell growth and tumor forming capacity. OPN is not mutationally activated in cancer but its expression is regulated by Wnt/Tcf signaling, steroid receptors, growth factors, ras, Ets and AP-1 transcription factors. Presumably these factors are implicated in induction of OPN overexpression in cancer. Greater understanding of the role of OPN in neoplastic change and its transcriptional regulation may enable development of novel cancer treatment strategies. PMID- 17113339 TI - A biplanar fluoroscopic approach for the measurement, modeling, and simulation of needle and soft-tissue interaction. AB - A methodology for modeling the needle and soft-tissue interaction during needle insertion is presented. The approach consists of the measurement of needle and tissue motion using a dual C-arm fluoroscopy system. Our dual C-arm fluoroscopy setup allows real time 3-D extraction of the displacement of implanted fiducials in the soft tissue during needle insertion to obtain the necessary parameters for accurate modeling of needle and soft-tissue interactions. The needle and implanted markers in the tissue are tracked during the insertion and withdrawal of the needle at speeds of 1.016 mm/s, 12.7 mm/s and 25.4 mm/s. Both image and force data are utilized to determine important parameters such as the approximate cutting force, puncture force, the local effective modulus (LEM) during puncture, and the relaxation of tissue. We have also validated the LEM computed from our finite element model with arbitrary needle puncture tasks. Based on these measurements, we developed a model for needle insertion and withdrawal that can be used to generate a 1-DOF force versus position profile that can be experienced by a user operating a haptic device. This profile was implemented on a 7-DOf haptic device designed in our laboratory. PMID- 17113340 TI - Overexpression of cyclin D1 and cortactin is primarily independent of gene amplification in salivary gland adenoid cystic carcinoma. AB - Adenoid cystic carcinoma (ACC) of the salivary glands exhibits persistent growth, invasion and metastasis. Chromosome 11q13 amplification is a frequent event associated with tumor progression in a number of carcinomas and is associated with poor prognosis. Two genes within the 11q13 amplicon that are overexpressed as a result of 11q13 amplification are the cell cycle regulatory protein cyclin D1 (CCND1) and cortactin (CTTN), a protein involved cell motility and invasion. To determine the expression and gene status of cyclin D1 and cortactin in ACC, we evaluated 39 ACC cases by immunohistochemistry (IHC) for cyclin D1 and cortactin expression. Amplification of CCND1 and CTTN was determined by fluorescent in situ hybridization (FISH). Cyclin D1 overexpression was present in 90% (35/39) and cortactin expression in 62% (24/39) of evaluated cases, although CCND1 and CTTN levels were elevated in only two cases (5%) as determined by FISH. Our results indicate that chromosome 11q13 amplification is uncommon in ACC, but that cyclin D1 and cortactin are frequently overexpressed and may therefore contribute to the growth and invasive potential of ACC. PMID- 17113341 TI - Investigations of EPR parameters for the trigonal Ti3+-Ti3+ pair in beryl crystal. AB - By using the complete diagonalization of energy matrix of 3d1 ions in trigonal symmetry, the EPR parameters (g factors g( parallel), g( perpendicular) and zero field splitting D) of the trigonal Ti3+-Ti3+ pair in beryl crystal are calculated. In the calculations, the exchange interaction in the Ti3+-Ti3+ pair is taken as the perturbation and the local trigonal distortion in the defect center is considered. The results (which are in agreement with the experimental values) are discussed. PMID- 17113342 TI - Synthesis and characterization of iron(III), manganese(II), cobalt(II), nickel(II), copper(II) and zinc(II) complexes of salicylidene-N anilinoacetohydrazone (H2L1) and 2-hydroxy-1-naphthylidene-N anilinoacetohydrazone (H2L2). AB - Salicylidene-N-anilinoacetohydrazone (H(2)L(1)) and 2-hydroxy-1-naphthylidene-N anilinoacetohydrazone (H(2)L(2)) and their iron(III), manganese(II), cobalt(II), nickel(II), copper(II) and zinc(II) complexes have been synthesized and characterized by IR, electronic spectra, molar conductivities, magnetic susceptibilities and ESR. Mononuclear complexes are formed with molar ratios of 1:1, 1:2 and 1:3 (M:L). The IR studies reveal various modes of chelation. The electronic absorption spectra and magnetic susceptibility measurements show that the iron(III), nickel(II) and cobalt(II) complexes of H(2)L(1) have octahedral geometry. While the cobalt(II) complexes of H(2)L(2) were separated as tetrahedral structure. The copper(II) complexes have square planar stereochemistry. The ESR parameters of the copper(II) complexes at room temperature were calculated. The g values for copper(II) complexes proved that the Cu-O and Cu-N bonds are of high covalency. PMID- 17113343 TI - Theoretical studies on NMR chemical shifts in azacubanes. AB - Successive substitution of CH groups of cubane (CH)(8), by isoelectronic nitrogen atoms leads to a class of energy-rich azacubanes (CH)(8-alpha)N(alpha) (with alpha=1-8). In the present work, we systematically investigate how substitution of nitrogen in a cubanoid influence deshielding of carbon and manifest in the chemical shift in NMR spectra calculated using the second order Moller-Plesset (MP2) perturbation level of theory. PMR spectra predict a large downshift of delta(H)=7.9 ppm in heptaazacubane owing to the more number of nitrogen and the stronger C-H...N interactions. These chemical shifts are explained by the net atomic charges derived from the population analysis based on Hirshfeld partitioning scheme. PMID- 17113344 TI - Raman spectroscopic study of the molecular structure of the uranyl mineral zippeite from Jachymov (Joachimsthal), Czech Republic. AB - Raman spectra at 298 and 77K and infrared spectra of the uranyl sulfate mineral zippeite from Jachymov (Joachimsthal), Czech Republic, K(0.6)(H(3)O)0.4[(UO(2))6(SO(4))3(OH)7].8H2O, were studied. Observed bands were tentatively attributed to the (UO(2))2+ and (SO(4))2- stretching and bending vibrations, the OH stretching vibrations of water molecules, hydroxyls and oxonium ions, and H(2)O, oxonium, and delta U-OH bending vibrations. Empirical relations were used for the calculation of U-O bond lengths in uranyl R (A)=f(nu(3) or nu(1)(UO(2))2+). Calculated U-O bond lengths are in agreement with U-O bond lengths from the single crystal structure analysis and those inferred for uranyl anion sheet topology of uranyl pentagonal dipyramidal coordination polyhedra. The number of observed bands supports the conclusion from single crystal structure analysis that at least two symmetrically distinct U6+ (in uranyls) and S6+ (in sulfates), water molecules and hydroxyls may be present in the crystal structure of the zippeite studied. Strong to very weak hydrogen bonds present in the crystal structure of zippeite studied were inferred from the IR spectra. PMID- 17113345 TI - Using digital videos displayed on personal digital assistants (PDAs) to enhance patient education in clinical settings. AB - OBJECTIVES: To evaluate the effects of using an audiovisual animation (i.e., digital video) displayed on a personal digital assistant (PDA) for patient education in a clinical setting. METHODS: Quasi-experimental study of a prospective technology intervention conducted in an outpatient infectious diseases clinic at an academic medical center. Subjects responded to questions immediately before, immediately after, and 4-6 weeks after watching a digital video on a PDA. Outcome measures include participant knowledge of disease, knowledge of medications, and knowledge of adherence behaviors; attitudes toward the video and PDA; self-reported adherence; and practicality of the intervention. RESULTS: Fifty-one English-speaking adults who were initiating or taking medications for the treatment of HIV/AIDS participated in the study. At visit one, statistically significant improvements in knowledge of disease (p<0.005; paired t-test), knowledge of medications (p<0.005; paired t-test), and knowledge of adherence behaviors (p<0.05; ANOVA) were measured after participants watched the PDA-based video. At visit two (4-6 weeks later), statistically significant improvements in self-reported adherence to the medication regimens (p<0.005; paired t-test) were reported. Participants liked the PDA-based video and indicated that it was an appropriate medium for learning, regardless of their baseline literacy skills. The video education process was estimated to take 25 min of participant time and was viewed in both private and semi-private locations. CONCLUSIONS: Technology-assisted education using a digital video delivered via PDA is a convenient and potentially powerful way to deliver health messages. The intervention was implemented efficiently with participants of a variety of ages and educational levels, and in a range of locations within clinical environments. Additional study of this methodology is warranted. PMID- 17113346 TI - Comparative antibacterial activity of polymeric 3-alkylpyridinium salts isolated from the Mediterranean sponge Reniera sarai and their synthetic analogues. AB - Metabolites from marine sponges are considered a promising alternative to heavy metals in antifouling coatings. Water-soluble polymeric 3-alkylpyridinium salts and 14 related synthetic analogues showed considerable antibacterial activity against marine biofilm bacteria and may represent good candidates as natural biocides for marine technology applications. PMID- 17113347 TI - A new platelet storage lesion index based on paired samples, without and with EDTA and cell counting: comparison of three types of leukoreduced preparations. AB - Platelets derived from whole blood and diverse apheresis procedures come in contact with various artificial surfaces and undergo contact activation, sheer stress-induced shape changes, aggregation and microvesiculation during collection, processing and storage. These dynamic changes are qualitatively reflected in the log-normal platelet size distribution patterns seen, when using modern automated cell counters and can be measured quantitatively by counting the paired samples, without and with added EDTA, and calculating the differences (d) in platelet cellular indices (dPLT/dMPV/dPDW). Reporting the differences instead of the absolute values of cellular indices makes the measurements independent of basic principles used for cell counting (i.e. aperture-impedance or flow-optic). The measurements can be performed either in blood centres, hospital blood banks or nearby patient clinics equipped with a validated cell counter. At least three useful quality indices could be derived simultaneously from this procedure (accurate estimation of platelet yields using the value of the EDTA-containing sample); quantitative assessment of the % of reversible platelet aggregates, an age/pH/temperature-dependent degree of microvesiculation/apoptosis/PS exposure, based on the response to EDTA. This new set of indices correlate significantly with other conventional tests for platelet quality; hence, provide additional supportive evidence for confirming the low pH values and the subjective poor swirling data occasionally seen with stored PC. In the following study, the Sysmex SE 9000 was successfully employed for estimation of platelet cellular indices comparing the platelet storage lesion index of three types of leukoreduced platelet concentrates in current practice. The relationship between this in vitro response to clinical outcome remains to be established. PMID- 17113349 TI - Both ramipril and telmisartan reverse indices of early diabetic cardiomyopathy: a comparative study. AB - AIMS: We tested the hypothesis that renin-angiotensin system inhibition could reverse left ventricular diastolic dysfunction in patients with type 2 diabetes. METHODS AND RESULTS: Forty asymptomatic patients with type 2 diabetes were recruited in this double-blind cross-over trial. Left ventricular diastolic function was assessed at baseline with Doppler echocardiography; ratios of early to late peak flow velocity through the mitral orifice (E/A) and velocity time integral of early to late transmitral diastolic flow (VTIE/VTIA) were evaluated. In addition, plasma brain natriuretic peptide (BNP) was measured. Patients received randomly either ramipril (2.5 mg/day), or telmisartan (40 mg/day) or their combination for 3 months. Subsequently, every patient was crossed over to alternative regimens after a 2-week washout period. Measurements were repeated at the end of each treatment period. Both E/A and VTIE/VTIA ratios were increased (29 and 20% with ramipril, 25 and 23% with telmisartan and 36 and 28% with combination treatment, respectively, p < 0.001), whereas plasma BNP levels were significantly reduced with all 3 regimens (9% with ramipril, 25% with telmisartan and 36% with combination, p < 0.001). CONCLUSIONS: Both ramipril and telmisartan improve echocardiographic left ventricular diastolic indices and reduce plasma BNP levels in diabetic patients; their combination yields an even better therapeutic effect. PMID- 17113348 TI - Novel tumor antigens elicit anti-tumor humoral immune reactions in a subset of patients with polycythemia vera. AB - We attempted to determine whether the immune reactions elicited by aberrantly expressed testis antigens contribute to the beneficial responses to interferon (IFN)-alpha therapy and other therapies in patients with polycythemia vera (PV). We screened a human testis cDNA library using SEREX (serological analysis of tumor antigens by screening an expression cDNA library with sera from three patients with PV who had undergone IFN-alpha-induced or other therapeutics induced remission). We identified two novel PV associated tumor antigens, PV65 (eIF-2alpha) and PV13 (protamine 2). These 2 antigens elicited IgG antibody reactions in a subset of PV patients but not in healthy donors, suggesting that they are authentic tumor antigens. Increased phosphorylation of PV65 in response to stimulation of IFN-alpha, and upregulation of PV13 in tumor cells might enhance their abilities in elicitation of immune reactions in patients. These findings provide new insights into the mechanism underlying the regulation of the self-antigen repertoire in eliciting anti-tumor immune reactions in patients with polycythemia vera, and suggest their potential as the targets of novel immunotherapy. PMID- 17113350 TI - Pain catastrophizing, response to experimental heat stimuli, and post-cesarean section pain. AB - This prospective study assessed the relation between pain catastrophizing, response to experimental pain stimuli, and pain perceived by women after elective cesarean sections. Forty-seven women who were scheduled for elective cesarean section were enrolled in the study. Magnitude estimation to suprathreshold phasic and tonic heat pain stimuli was assessed 1 or 2 days before surgery. Women completed the Pain Catastrophizing Scale after the heat stimuli and again on the first postoperative day. During the first and second postoperative days, perception of pain intensity was assessed by visual analog scale at each analgesia request. A multiple regression analysis revealed that pain on the first postoperative day was predicted by patient response to preoperative tonic heat stimuli (r(2) = .167, P = .008). Pain on the second postoperative day was predicted by preoperative pain catastrophizing (r(2) = .139, P = .021). No significant association was observed between preoperative response to heat stimuli or pain catastrophizing and the patient's analgesic consumption in the obstetrical ward. It is concluded that pain catastrophizing and response to experimental tonic heat pain correlate with post-cesarean section pain. PERSPECTIVE: This article presents psychological and psychophysical measures that may be of help in the prediction of post-cesarean section pain. It may therefore contribute to the treatment of the sequelae of the most common major surgical procedure performed in women in their reproductive years. PMID- 17113351 TI - Relationship between the firing frequency of injured peripheral neurons and inhibition of firing by sodium channel blockers. AB - Animal models of neuropathic pain in which a peripheral nerve is damaged result in spontaneous activity in primary afferents that can be inhibited by intravenous administration of sodium channel blockers. Many of these compounds exhibit use dependent block of sodium current, leading to the prediction that they should more readily inhibit neurons that fire at higher frequencies. This prediction was tested in 2 rat models of nerve injury, L5 spinal nerve section and sciatic nerve section. Sciatic nerve section produced average firing frequencies that were higher than spinal nerve section and often manifested as high-frequency bursting. Inhibition of firing by intravenous sodium channel blockers was longer lasting in this model. Within each model, higher frequency of firing did not translate into more effective block. In the spinal nerve section model, there was a robust inverse correlation between frequency and inhibition. Within the sciatic section model, only neurons that fired in rhythmic bursts were inhibited, and again, those firing at lower mean frequencies were more effectively inhibited. These results indicate that the efficacy of sodium channel blockers depends on the nature of the injury and the pattern of the resulting activity rather than simply the frequency of action potentials generated. PERSPECTIVE: This study examines the ability of frequency-dependent sodium channel blockers to inhibit spontaneous firing of injured peripheral nerves in vivo. It outlines the conditions under which inhibition is more and less effective and will provide insight into conditions under which sodium channel blockers are likely to be therapeutically useful. PMID- 17113352 TI - Critical evaluation of the colocalization between calcitonin gene-related peptide, substance P, transient receptor potential vanilloid subfamily type 1 immunoreactivities, and isolectin B4 binding in primary afferent neurons of the rat and mouse. AB - Calcitonin gene-related peptide (CGRP) and/or substance P (SP) immunoreactivity as well as isolectin B(4) (IB(4)) binding are commonly used to define peptidergic and non-peptidergic nociceptor populations, respectively. Although this demarcation is well supported in the mouse, there is accumulating evidence to suggest it is not so in the rat. Hence, this investigation was undertaken to evaluate and quantify the colocalization of the neuropeptides CGRP and SP with IB(4) binding sites and the transient receptor potential vanilloid subfamily type 1 (TRPV1) channel and to compare this colocalization between trigeminal (TG) and dorsal root ganglia (DRG) in adult rats. These findings illustrate that there is a substantial overlap ( approximately 45% in the DRG and approximately 30% in the TG) between peptidergic neurons (ie, CGRP- and SP-expressing) and neurons that bind IB(4) in rat sensory ganglia. However, there were also significant differences in the colocalization of these markers between the DRG and TG. For instance, in the DRG, significantly more CGRP-immunoreactive neurons also expressed IB(4) binding sites (44.5%) compared with the TG (27.5%). In contrast, significantly fewer CGRP-immunoreactive neurons in the DRG colocalized TRPV1 immunoreactivity (49.2%) compared with the TG (70%). Moreover, we directly assessed the colocalization of CGRP and IB(4) in the TG of rats and mice using a CGRP antibody that recognizes this peptide in both species. Thus, whereas only an approximately 10% overlap was observed in TG neurons of mouse, significantly greater overlap (approximately 35%) was observed in those of rat. PERSPECTIVE: These data indicate that in adult rat sensory ganglia, there is not a clear distinction between the peptidergic and non-peptidergic nociceptor subclasses as a function of IB(4) binding. Furthermore, there are significant differences between the TG and DRG in the degree to which commonly utilized nociceptive neuronal markers are co-expressed. Taken together, the present findings dictate prudence when extrapolating experimental conclusions about the neurochemical classification of neurons between sensory ganglia or between species, including humans. PMID- 17113353 TI - Placebo-controlled comparison of a morphine/dextromethorphan combination with morphine on experimental pain and hyperalgesia in healthy volunteers. AB - In this double-blind, placebo-controlled, crossover study we compared the analgesic effect of a single oral dose of 30-mg dextromethorphan and 30-mg morphine combination (MS/DM) to 30 mg morphine (MS) alone and either placebo or 30 mg dextromethorphan (DM) on cutaneous sensitization induced by heat/capsaicin (topical) sensitization on the forearm and the brief thermal sensitization model on the thigh in 22 healthy volunteers. Outcome measures were areas of secondary hyperalgesia to brush and von Frey hair stimulation in both sensitization models and the painfulness of acute thermal noxious stimulation on the upper arm. Compared with placebo, both MS/DM and morphine had some effect on the secondary hyperalgesia and reduced the painfulness of a noxious thermal stimulus. The analgesic effect of MS/DM was not superior to that of morphine on any outcome measure. These results differ from preclinical studies with animal experimental pain models in which DM markedly potentiated the analgesic effects of opioids, but they are in accordance with recent clinical trials for chronic pain. PERSPECTIVE: Adding dextromethorphan to morphine (1:1 ratio) did not enhance analgesia on measures of experimental cutaneous sensitization and acute noxious thermal stimulation in healthy volunteers. The results differ from preclinical studies but agree with clinical trials. Human experimental models of pain and neuronal sensitization, which are responsive to oral opioids, allow efficient study of opioid combination analgesics and simplify the process for determining the optimal dose and/or dose ratio. PMID- 17113354 TI - Importance of metallothioneins in the cadmium detoxification process in Daphnia magna. AB - Good knowledge of the relationship between toxic metals and biological systems, particularly the sub-cellular fraction, could be a suitable early indicator of toxic effects. These effects and the sub-cellular behaviour of cadmium were studied with a widely used species in freshwater toxicity bioassays, Daphnia magna. In spite of this very commonplace usage in ecotoxicological studies, very few data are available on its toxicant metabolism and in particular metal homeostasis. Combining multi-tools analysis, a soluble protein was found: it is heat-stable, rich in sulfhydryl groups (differential pulse polarography), characterised by a molecular mass of approximately 6.5 kDa, with a G-75 chromatographic profile corresponding to the rabbit metallothioneins monomer, with few if any aromatic-containing amino acids, it binds metals (e.g. Cd, Cu), and its concentration increases with Cd exposure. This evidence led us to hypothesise that metallothioneins (MTs) are present in D. magna. Up to 75% of the Cd body burden with Cd exposure is bound to the MTs fraction. The increase in the Cd concentration in the surrounding medium and concomitantly in daphnids induces sub-cellular reorganisation of essential metals such as Cu and Zn. The rate of metals in the soluble cellular fraction and associated with MTs increases with the Cd body burden. Monitoring sub-cellular distribution of metals after exposure in the natural environment could be very useful for ecotoxicological assessment. PMID- 17113355 TI - Ibuprofen or ozagrel increases NO release and l-nitro arginine induces TXA(2) release from cultured porcine basilar arterial endothelial cells. AB - The vascular resting tone of the porcine basilar artery appears to be mostly maintained by a balance between spontaneously released nitric oxide (NO) from endothelial cells and thromboxane (TX) A(2) from endothelial and smooth muscle cells. However the precise role of the interaction between the above two substances in the control of vascular tone is unclear. We attempted to clarify the interaction between NO and TXA(2) using cultured porcine basilar arterial endothelial cells. The cultured endothelial cells produced NO spontaneously, while TXB(2) (a stable metabolite of TXA(2)) production remained below the detection limit. Ibuprofen (a COX inhibitor) and ozagrel (a TXA(2) synthetase inhibitor) significantly increased the spontaneous production of NO, which was not affected by 1400W (an iNOS inhibitor). l-Nitro arginine (a NOS inhibitor) significantly induced TXB(2) production. These results suggest that NO may inhibit COX or TXA(2) synthetase, and that therefore inhibition of NOS might disinhibit COX or TXA(2) synthetase, subsequently inducing TXA(2) production. On the other hand, as TXA(2) and other contractility-related prostaglandin(s) may inhibit NOS, therefore the inhibition of COX or TXA(2) synthetase might disinhibit NOS, and then increase the spontaneous production of NO in porcine basilar arterial endothelial cells. PMID- 17113356 TI - Inhibition of Src family tyrosine kinases prevents lipopolysaccharide-induced hyporeactivity in isolated rat tail arteries. AB - Tyrosine kinases may play a role in the vascular response to sepsis. We investigated the effect of selective inhibitors of Src family tyrosine kinases (SFK) on lipopolysaccharide (LPS)-induced vascular hyporeactivity. Rat tail artery segments were mounted in an isometric wire myograph. The effect of incubation with LPS was examined on phenylephrine (PE) and high potassium (KPSS) induced contraction, with and without the selective SFK inhibitors SU6656 or PP1. Western blotting was performed to assess SFK phosphorylation and iNOS induction. Incubation with LPS for 18 h induced marked vascular hyporeactivity to both PE (p<0.001) and KPSS (P<0.001). Incubation with SU6656 alone had no effect on contractility to PE and KPSS, and SU6656 partially prevented LPS-induced hyporeactivity to PE (p<0.01) and KPSS (p<0.001). In contrast, PP1 alone diminished contractility to PE (p<0.01) and KPSS (p<0.001), and co-incubation of LPS with PP1 completely prevented LPS-induced hyporeactivity. LPS increased tyrosine phosphorylation of SFK and this effect was inhibited by SFK inhibitors. LPS also increased levels of iNOS and this was also inhibited by SU6656 and PP1. LPS-induced hyporeactivity in vitro is mediated by activation of SFK. Selective inhibitors of SFK may have therapeutic potential in the management of septic shock. PMID- 17113357 TI - Phase II multi-step planning methods in oncology: comparison, recommendations and potential applications. AB - Phase II clinical trials in oncology are commonly used to determine whether a new treatment has a sufficient response rate to be compared with the best standard therapy in phase III. The multi-step planning methods such as Fleming's procedure and the triangular test adapted for phase II trials were elaborated to terminate phase II trials early. We compared these methods considering a fixed number of steps (4 or 5) on their statistical properties: overall type II error and observed type II error, average number of subjects necessary to conclude, probability to conclude at each step and maximum number of subjects necessary to conclude. Fleming's procedure has an actual power similar to the theoretical power for low to high response rate. In contrast, triangular test has an actual power different from the theoretical power in a few situations: it was very high in case of a very low minimum response rate and very low (<20%) in case of a very high minimum response rate (>80%). In these situations, Fleming's procedure and triangular test were not compared and triangular test cannot be recommended. For intermediate response rate, triangular test required a lower average number of subjects to conclude, a larger number of subjects at each step and thus a larger maximum number of subjects. It provided a larger probability to conclude during the first steps. These advantages should be balanced with the risk to have to include a larger number of patients. Multi-step methods, when correctly used are useful for cytotoxic development when response rate is the end point. They can also be used in trials where toxicity is the end point, and could be of great interest for cytostatic development for example with biological surrogate endpoints. An example using real phase II data is also presented. PMID- 17113358 TI - Autonomic pathways regulating pancreatic exocrine secretion. AB - The parasympathetic (PNS) and sympathetic (SNS) and nervous systems densely innervate the exocrine pancreas. Efferent PNS pathways, consisting of central dorsal motor nucleus of the vagus (DMV) and peripheral pancreatic neurons, stimulate exocrine secretion. The DMV integrates cortical (olfactory, gustatory) and gastric, and intestinal vagal afferent input to determine central PNS outflow during cephalic, gastric and intestinal phases of exocrine secretion. Pancreatic neurons integrate DMV input with peripheral enteric, sympathetic, and, possibly, afferent axon reflexes to determine final PNS input to all exocrine effectors. Gut and islet hormones appear to modulate both central and peripheral PNS pathways. Preganglionic sympathetic neurons in the intermediolateral (IML) column of the spinal cord receive inputs from brain centers, some shared with the PNS, and innervate postganglionic neurons, mainly in prevertebral ganglia. Sympathetic innervation of the exocrine pancreas is primarily indirect, and inhibits secretion by decreasing blood flow and inhibiting transmission in pancreatic ganglia. Interactions between SNS and PNS pathways appear to occur in brain, spinal cord, pancreatic and prevertebral ganglia, and at neuroeffector synapses. Thus, the PNS and SNS pathways regulating the exocrine pancreas are directly or indirectly antagonistic at multiple sites: the state of exocrine secretion reflects the balance of these influences. Despite over a century of study, much remains to be understood about the connections of specific neurons forming pancreatic pathways, their processes of neurotransmission, and how disruption of these pathways contributes to pancreatic disease. PMID- 17113359 TI - Voltammetric determination of ethinylestradiol at a carbon paste electrode in the presence of cetyl pyridine bromine. AB - Electrochemical behaviors of ethinylestradiol at a carbon paste electrode (CPE) in the presence of cetyl pyridine bromide (CPB) are investigated by electrochemical techniques. Compared with that at a CPE without CPB, the oxidation peak potential of ethinylestradiol shifts negatively and the peak current is increased significantly, due to the enhanced accumulation of ethinylestradiol via electrostatic interaction with CPB at the hydrophobic electrode surface. It is verified by the influences of different kinds of surfactants on the electrochemical signals of ethinylestradiol. Some parameters such as pH, scan rate, accumulation potential and accumulation time on the oxidation of ethinylestradiol are optimized. Under optimal conditions, the oxidation peak current is proportional to ethinylestradiol concentration in the range of 5.0 x 10(-8) to 2.0 x 10(-5) mol L(-1) with a detection limit of 3.0 x 10(-8) mol L(-1) for 150 s accumulation by linear sweep voltammetry (LSV). The proposed procedure is successfully applied to determine ethinylestradiol in pharmaceutical formulation (Levonorgestrel and Etinylestradiol tablets) and the results are satisfying compared with that of high-performance liquid chromatography (HPLC). PMID- 17113360 TI - Impedance sensing of allergen-antibody interaction on glassy carbon electrode modified by gold electrodeposition. AB - The interactions between the recombinant dust mite allergen Der f2 and murine monoclonal antibody were monitored by electrochemical impedance spectroscopy (EIS). Allergen Der f2 were immobilized through the nanogold formed by electrodeposition of gold on planar glassy carbon electrode. A 30-s gold electrodeposition provided a desirable substrate for the immobilization of allergen. Electrochemical deposition of gold on a glassy carbon electrode showed significant improvement in allergen immobilization. The impedance measurements were based on the charge-transfer kinetics of the [Fe(CN)(6)](3-/4-) redox pair. The interactions between allergen and antibody occurred on electrode surface altered the interfacial electron transfer resistance, R(CT), by preventing the redox species approaching the electrode. The results showed that R(CT) increased with increasing concentration of monoclonal antibodies. PMID- 17113361 TI - Voltammetric determination of catalytic reaction parameters of laccase based on electrooxidation of hydroquinone and ABTS. AB - A convenient method for the measurement of the catalytic activity of laccase is proposed based on the voltammetric determination of catalytic reaction substrates: 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate) diammonium salt (ABTS) and 1,4-benzenediol (hydroquinone). The measurement performed using microelectrodes working under spherical diffusion conditions is both accurate and simple, and allows to monitor parallely the consumption of substrate and formation of product of the catalytic reaction. The method proposed in this paper was compared with the two generally employed procedures based on oxygen measurement by Clark electrode and on spectrophotometry. The procedure described in the present paper was found to be simpler and more reproducible results were obtained than using Clark electrode. Compared to spectrophotometry a larger range of catalytic reaction substrates can be studied including colorless compounds. PMID- 17113362 TI - The F(1)F(0) ATP synthase and mitochondrial respiratory chain complexes are present on the plasma membrane of an osteosarcoma cell line: An immunocytochemical study. AB - F(1)F(0) ATP synthase is ectopically expressed on the surface of several cell types, including endothelium and cancer cells. This study uses immunocytochemical detection methods via highly specific monoclonal antibodies to explore the possibility of plasma membrane localization of other mitochondrial proteins using an osteosarcoma cell line in which the location of the mitochondrial reticulum can be clearly traced by green fluorescent protein tagging of the organelle. We found that subunits of three of the four respiratory chain complexes were present on the surface of these cells. Additionally, we show for the first time that F(0) subunits d and OSCP of the ATP synthase are ectopically expressed. In all cases the OXPHOS proteins show a punctate distribution, consistent with data from proteome analysis of isolated lipid rafts that place the various mitochondrial proteins in plasma membrane microdomains. We also examined the cell surface for marker membrane proteins from several other intracellular organelles including ER, golgi and nuclear envelope. They were not found on the surface of the osteosarcoma cells. We conclude that mitochondrial membrane proteins are ectopically expressed, but not proteins from other cellular organelles. A specific mechanism by which the mitochondrion and plasma membrane fuse to deliver organellar proteins is suggested. PMID- 17113363 TI - Non-invasive measurement of total respiratory compliance and resistance in cats. AB - We adapted non-invasive techniques developed for human infants to measure total respiratory system compliance (Crs) and resistance (Rrs) in 21 healthy cats. The animals breathed through a face mask attached to a respiratory circuit and measurements were taken of changes in lung volume and airway pressure during brief occlusions of the airway at different lung volumes. The slope of the plot of change in volume against airway pressure yielded the multiple occlusion Crs with a mean (+/-95%CI) value of 6.8 (6.3-7.3) ml/cm H2O. In 12 animals measurements were made by the single breath technique in which occlusion was made early in expiration and on release, a plot of the subsequent relaxed expiratory flow and volume yielded the time constant (taurs), Crs and Rrs with mean (+/ 95%CI) values of 0.27 (0.22-0.31) s, 7.0 (6.1-7.8) ml/cm H2O, and 38.7 (33.7 43.6) cm H2O/l/s, respectively. Rrs was significantly correlated inversely with forced expiratory flow at resting lung volume (V'maxFRC). PMID- 17113364 TI - Prenatal nicotine exposure transiently alters the lung mechanical response to hypoxia in young lambs. AB - To test the hypothesis that fetal nicotine exposure alters the lung mechanical response to hypoxia (10% O(2)) 10 lambs were exposed during the last fetal trimester to a low dose nicotine (LN) and 10 to a moderate dose (MN) (maternal dose 0.5 and 1.5mg/(kgday) free base, respectively). There were 10 controls (C). At 12 days, minute ventilation increased significantly less in MN compared with LN but not with C. In contrast to C and LN, MN did not show anticipated increases in dynamic compliance, specific compliance and FRC or decrease in lung resistance but had signs of airway hyperreactivity during hypoxia. Nicotine exposure did not alter the cardiovascular response. These adverse effects decreased with advancing age. In summary, prenatal nicotine exposure alters the lung mechanical response to hypoxia. We speculate that prenatal nicotine-induced alterations of lung mechanics during hypoxia may contribute to an increased vulnerability to hypoxic stress during infancy. PMID- 17113365 TI - Quantitative determination of donepezil in human plasma by liquid chromatography/tandem mass spectrometry employing an automated liquid-liquid extraction based on 96-well format plates. Application to a bioequivalence study. AB - An automated high-throughput liquid chromatography/tandem mass spectrometry (LC MS/MS) method was developed for quantitative determination of donepezil in human plasma. 150 MicroL of plasma samples were placed in 2.2 mL 96-deepwell plates and both donepezil and loratadine (IS) were extracted from human plasma by liquid liquid extraction (LLE), using hexane as the organic solvent. Robotic liquid handling workstations were employed for all liquid transfer and solution preparation steps and resulted in a short sample preparation time. After vortexing, centrifugation and freezing, the supernatant organic solvent was evaporated and reconstituted in a small volume of reconstitution solution. The method developed, includes a sample analysis performed by reversed phase LC MS/MS, with positive ion electrospray ionization, using multiple reaction monitoring (MRM). The chromatographic run time was set for 2.0 min with a flow rate of 0.7 mL/min in a C18 analytical column. The method was significantly sensitive, specific, accurate and precise for the determination of donepezil in human plasma and had the shortest run time. The curve was proven to be linear for the concentration range of 0.1-100 ng/mL. After validation, the method was applied to the rapid and reliable quantitative determination of donepezil in a bioequivalence study after per os administration of a 5mg donepezil tablet. PMID- 17113366 TI - Combining poly (methacrylic acid-co-ethylene glycol dimethacrylate) monolith microextraction and on-line pre-concentration-capillary electrophoresis for analysis of ephedrine and pseudoephedrine in human plasma and urine. AB - A method based on poly (methacrylic acid-co-ethylene glycol dimethacrylate) (MAA EGDMA) monolith microextraction (PMME) and field-enhanced sample injection (FESI) pre-concentration technique was proposed for sensitive capillary electrophoresis ultraviolet (CE-UV) analysis of ephedrine (E) and pseudoephedrine (PE) in human plasma and urine. The PMME device consisted of a regular plastic syringe (1 mL), a poly (MAA-EGDMA) monolithic capillary (2 cm x 530 microm I.D.) and a plastic pinhead connecting the former two components seamlessly. The extraction was achieved by driving the sample solution through the monolithic capillary tube using a syringe pump, for the desorption step, an aliquot of organic solvent, which normally provided an excellent medium to ensure direct compatibility for FESI in CE, was injected via the monolithic capillary and collected into a vial for subsequent analysis by CZE. The best separation was achieved using a buffer composed of 0.1M phosphate electrolyte (pH 2.5) and 10% acetonitrile (v/v). The combination of both pre-concentration procedures allowed the detection limits of the analytes down to 5.3 ng/mL and 8.0 ng/mL in human plasma and urine, respectively. Excellent method of reproducibility was found over a linear range 50-5000 ng/mL in plasma and urine sample. Plasma and urine samples from volunteers receiving pseudoephedrine have also been successfully analysed. PMID- 17113367 TI - Cation exchange chromatography in antibody purification: pH screening for optimised binding and HCP removal. AB - The production of pharmaceutical antibodies requires reliable and rapid processes with high purity and yield. Although protein A gels selectively and efficiently bind antibodies in the capture step, intense research is going on to find alternatives that can abolish the drawbacks of protein A chromatography. Ion exchangers e.g. are more robust, considerably cheaper and can eliminate ligand leaching. For the strong cation exchangers Fractogel EMD SO3- (M) and Fractogel EMD SE Hicap (M) we have evaluated the influence of pH for optimised binding and removal of host cell protein (HCP). In a fast initial screening we measured batch binding capacities. Subsequent scale-down to 96-well plate format proved that assay miniaturisation still provided reliable data. We demonstrated with the principle of residence time that scout columns are suitable for dynamic studies. The optimum pH range from batch binding was transferred to scout columns which were then used to screen for maximum dynamic capacities. In addition IEF titration curve analysis was employed to define a final operational pH. With this pH we ran labscale columns to purify monoclonal antibody. The cation exchangers showed high step yields and host cell proteins in the pools from gradient elution were reduced very effectively. PMID- 17113368 TI - Affinity chromatography: a useful tool in proteomics studies. AB - Separation or fractionation of a biological sample in order to reduce its complexity is often a prerequisite to qualitative or quantitative proteomic approaches. Affinity chromatography is an efficient protein separation method based on the interaction between target proteins and specific immobilized ligands. The large range of available ligands allows to separate a complex biological extract in different protein classes or to isolate the low abundance species such as post-translationally modified proteins. This method plays an essential role in the isolation of protein complexes and in the identification of protein-protein interaction networks. Affinity chromatography is also required for quantification of protein expression by using isotope-coded affinity tags. PMID- 17113369 TI - Jumbo squid beaks: inspiration for design of robust organic composites. AB - The hard tissues found in some invertebrate marine organisms represent intriguing paradigms for robust, lightweight materials. The present study focuses on one such tissue: that comprising the beak of the jumbo squid (Dosidicus gigas). Its main constituents are chitin fibers (15-20wt.%) and histidine- and glycine-rich proteins (40-45%). Notably absent are mineral phases, metals and halogens. Despite being fully organic, beak hardness and stiffness are at least twice those of the most competitive synthetic organic materials (notably engineering polymers) and comparable to those of Glycera and Nereis jaws. Furthermore, the combination of hardness and stiffness makes the beaks more resistant to plastic deformation when in contact with blunt abrasives than virtually all metals and polymers. The 3,4-dihydroxy-l-phenylalanine and abundant histidine content in the beak proteins as well as the pigmented hydrolysis-resistant residue are suggestive of aromatic cross-linking. A high cross-linking density between the proteins and chitin may be the single most important determinant of hardness and stiffness in the beak. Beak microstructure is characterized by a lamellar arrangement of the constituents, with a weak interface that promotes crack deflection and endows the structure with high fracture toughness. The susceptibility of this microstructure to cracking along these interfaces from contact stresses at the external surface is mitigated by the presence of a protective coating. PMID- 17113370 TI - Spotted hyenas. PMID- 17113371 TI - Digital cows grazing on digital grounds. AB - Picture a pasture open to all (...) As a rational being, each herdsman seeks to maximize his gain. Explicitly or implicitly, more or less consciously, he asks, "What is the utility to me of adding one more animal to the herd?" This utility has one negative and one positive component. 1) The positive component is a function of the increment of one animal. Since the herdsman receives all the proceeds from the sale of the additional animal, the positive utility is nearly +1. 2) The negative component is a function of the additional overgrazing created by one more animal. Since, however, the effects of overgrazing are shared by all the herdsmen, the negative utility for any particular decision-making herdsman is only a fraction of -1. Adding together the component partial utilities, the rational herdsman concludes that the only sensible course for him to pursue is to add another animal to the herd. And another; and another... PMID- 17113372 TI - There's more to magic than meets the eye. PMID- 17113373 TI - Deep brain stimulation of the subthalamic nucleus: a two-edged sword. PMID- 17113374 TI - Axon guidance: ephrins at WRK on the midline. AB - Recent findings indicate that the embryonic motor neurons act as gatekeepers to regulate midline crossing during development of the nematode Caenorhabditis elegans. The newly identified protein WRK-1 and ephrins cooperate to prevent longitudinal axons from crossing the midline. PMID- 17113375 TI - DNA repair: corrections in the golden years. AB - Genetic changes increase with the age of organisms, but the basis for this increase is unclear. A study has found that the major pathway of DNA repair is altered with age in the testes of male Drosophila, thus providing a powerful system to dissect the basis for age-related genomic changes. PMID- 17113376 TI - Evolution: the ecological reverberations of toxic trace elements. AB - A recent study of plants that accumulate selenium from soils illustrates how plant defenses can be sequestered and presumably exploited defensively by herbivores that have co-evolved selenium resistance. PMID- 17113377 TI - Cooperation: integrating evolutionary and ecological perspectives. AB - Putting a competitive squeeze on a cooperative group has long been considered to encourage cheats. Now we learn that competition, by driving diversification among cooperators, can create groups that are both more productive and more resistant to defection. PMID- 17113378 TI - Cognitive neuroscience: rewired or crosswired brains? AB - A new study of congenitally blind participants has provided important insights into the neuronal mechanisms of brain reorganization after injury, with implications for our knowledge of other cross-modal phenomena, such as synaesthesia, and for the generation of qualia. PMID- 17113379 TI - Disease pathology: wasting energy fighting infection. AB - Drosophila melanogaster infected with Mycobacterium marinum suffer metabolic wasting similar to that seen in humans suffering from tuberculosis. This wasting is linked to insulin signaling and hastens host death. PMID- 17113380 TI - Germline specification: small things have a big role. AB - Germline cell fate is specified by localized OSK, VAS and other components in the pole plasm of the Drosophila embryo. New work shows that a PIWI-mediated miRNA pathway contributes to this process by regulating OSK and VAS localization. PMID- 17113381 TI - Circadian rhythms: perturbing a food-entrained clock. AB - When food is scarce, a food-entrainable circadian clock coordinates mammalian activity rhythms with a predictable daily mealtime. Neural and molecular substrates of this circadian function have long eluded localization, but new studies suggest a critical role for a familiar circadian clock gene. PMID- 17113382 TI - Selenium-tolerant diamondback moth disarms hyperaccumulator plant defense. AB - BACKGROUND: Some plants hyperaccumulate the toxic element selenium (Se) to extreme levels, up to 1% of dry weight. The function of this intriguing phenomenon is obscure. RESULTS: Here, we show that the Se in the hyperaccumulator prince's plume (Stanleya pinnata) protects it from caterpillar herbivory because of deterrence and toxicity. In its natural habitat, however, a newly discovered variety of the invasive diamondback moth (Plutella xylostella) has disarmed this elemental defense. It thrives on plants containing highly toxic Se levels and shows no oviposition or feeding deterrence, in contrast to related varieties. Interestingly, a Se-tolerant wasp (Diadegma insulare) was found to parasitize the tolerant moth. The insect's Se tolerance mechanism was revealed by X-ray absorption spectroscopy and liquid chromatography-mass spectroscopy, which showed that the Se-tolerant moth and its parasite both accumulate methylselenocysteine, the same form found in the hyperaccumulator plant, whereas related sensitive moths accumulate selenocysteine. The latter is toxic because of its nonspecific incorporation into proteins. Indeed, the Se-tolerant diamondback moth incorporated less Se into protein. Additionally, the tolerant variety sequestered Se in distinct abdominal areas, potentially involved in detoxification and larval defense to predators. CONCLUSIONS: Although Se hyperaccumulation protects plants from herbivory by some invertebrates, it can give rise to the evolution of unique Se-tolerant herbivores and thus provide a portal for Se into the local ecosystem. In a broader context, this study provides insight into the possible ecological implications of using Se-enriched crops as a source of anti-carcinogenic selenocompounds and for the remediation of Se-polluted environments. PMID- 17113383 TI - Spatial and temporal control of cofilin activity is required for directional sensing during chemotaxis. AB - BACKGROUND: Previous work has led to the hypothesis that cofilin severing, as regulated by PLC, is involved in chemotactic sensing. We have tested this hypothesis by investigating whether activation of endogenous cofilin is spatially and temporally linked to sensing an EGF point source in carcinoma cells. RESULTS: We demonstrate that inhibition of endogenous cofilin activity with either siRNA or overexpression of LIMK suppresses directional sensing in carcinoma cells. LIMK siRNA knockdown, which suppresses cofilin phosphorylation, and microinjection of S3C cofilin, a cofilin mutant that is constitutively active and not phosphorylated by LIMK, also inhibits directional sensing and chemotaxis. These results indicate that phosphorylation of cofilin by LIMK, in addition to cofilin activity, is required for chemotaxis. Cofilin activity concentrates rapidly at the newly formed leading edge facing the gradient, whereas cofilin phosphorylation increases throughout the cell. Quantification of these results indicates that the amplification of asymmetric actin polymerization required for protrusion toward the EGF gradient occurs at the level of cofilin but not at the level of PLC activation by EGFR. CONCLUSIONS: These results indicate that local activation of cofilin by PLC and its global inactivation by LIMK phosphorylation combine to generate the local asymmetry of actin polymerization required for chemotaxis. PMID- 17113384 TI - Coordinated control of cell adhesion, polarity, and cytoskeleton underlies Hox induced organogenesis in Drosophila. AB - BACKGROUND: Hox genes control animal body plans by directing the morphogenesis of segment-specific structures. As transcription factors, HOX proteins achieve this through the activation of downstream target genes. Much research has been devoted to the search for these targets and the characterization of their roles in organogenesis. This has shown that the direct targets of Hox activation are often transcription factors or signaling molecules, which form hierarchical genetic networks directing the morphogenesis of particular organs. Importantly, very few of the direct Hox targets known are "realizator" genes involved directly in the cellular processes of organogenesis. RESULTS: Here, we describe for the first time a complete network linking the Hox gene Abdominal-B to the realizator genes it controls during the organogenesis of the external respiratory organ of the larva. In this process, Abdominal-B induces the expression of four intermediate signaling molecules and transcription factors, and this expression results in the mosaic activation of several realizator genes. The ABD-B spiracle realizators include at least five cell-adhesion proteins, cell-polarity proteins, and GAP and GEF cytoskeleton regulators. Simultaneous ectopic expression of the Abd-B downstream targets can induce spiracle-like structure formation in the absence of ABD-B protein. CONCLUSION: Hox realizators include cytoskeletal regulators and molecules required for the apico-basal cell organization. HOX-coordinated activation of these realizators in mosaic patterns confers to the organ primordium its assembling properties. We propose that during animal development, Hox-controlled genetic cascades coordinate the local cell-specific behaviors that result in organogenesis of segment-specific structures. PMID- 17113385 TI - Ritualized submission and the reduction of aggression in an invertebrate. AB - Ritualized behaviors that signify acceptance of a dominance relationship and reduce aggression between rivals are a common feature of vertebrate social behavior. Although some invertebrates, including crayfish, lobsters, and ants, display dominance postures, more complex dominance rituals and their effects on fitness have not been reported. We found that crayfish display such a complex ritual, when two males engaged in pseudocopulatory behavior to signify their dominance relationship. This was followed by a reduction in aggression and an increased likelihood of the subordinate's survival. Pseudocopulation was initiated by the eventual dominant and could be accepted or refused by the eventual subordinate. The frequency of aggressive behavior declined significantly during the first hour in all pairs that pseudocopulated but remained high in pairs that did not. Whereas all the subordinate members of pairs that pseudocopulated survived the initial 24 hr of pairing, half of subordinates that did not pseudocopulate were killed during that time. This differential mortality indicates that the reduction of aggression induced by the pseudocopulatory ritual directly enhances the differential survival of male crayfish that engage in this behavior. PMID- 17113386 TI - Element 1360 and RNAi components contribute to HP1-dependent silencing of a pericentric reporter. AB - In eukaryotes, distinct regions of the genome are packaged as euchromatin (less condensed, more active) or heterochromatin (condensed, silenced). Studies in yeast, plants, and flies suggest that RNA interference (RNAi) is linked to heterochromatin formation and transcriptional silencing of transposable element (TE) sequences. We previously reported that insertion of a mobile hsp70-white reporter within 10 kb of a 1360 element on chromosome four of Drosophila melanogaster correlates with variegation (silencing). Here, we report small RNAs (approximately 23 nt) corresponding to 1360, indicating processing by the RNAi machinery. To directly test the ability of 1360 to silence a nearby gene in vivo, we introduced a P element construct carrying a single copy of 1360 upstream of the hsp70-white reporter into flies. This 1360 element contributes to HP1 dependent variegation at a pericentric insertion site, as demonstrated by a decrease in silencing after FLP-mediated removal of 1360. In euchromatin, 1360 is not sufficient to induce silencing, suggesting that proximity to pericentric heterochromatin and/or a high local TE density contributes to heterochromatin formation. Silencing of the 1360, hsp70-white reporter is sensitive to mutations in RNAi components. Our results implicate 1360 as a target for sequence-specific heterochromatic silencing through an RNAi-dependent mechanism. PMID- 17113387 TI - Male chimpanzees prefer mating with old females. AB - Cross-cultural studies indicate that women's sexual attractiveness generally peaks before motherhood and declines with age. Cues of female youth are thought to be attractive because humans maintain long-term pair bonds, making reproductive value (i.e. future reproductive potential) particularly important to males. Menopause is believed to exaggerate this preference for youth by limiting women's future fertility. This theory predicts that in species lacking long-term pair bonds and menopause, males should not exhibit a preference for young mates. We tested this prediction by studying male preferences in our closest living relative, the chimpanzee (Pan troglodytes). We show that despite their promiscuous mating system, chimpanzee males, like humans, prefer some females over others. However, in contrast to humans, chimpanzee males prefer older, not younger, females. These data robustly discriminate patterns of male mate choice between humans and chimpanzees. Given that the human lineage evolved from a chimpanzee-like ancestor, they indicate that male preference for youth is a derived human feature, likely adapted from a tendency to form unusually long term mating bonds. PMID- 17113388 TI - CK2 controls the recruitment of Wnt regulators to target genes in vivo. AB - Nuclear beta-catenin is a transcriptional coactivator of LEF-1/TCF DNA-binding proteins in the Wnt/Wg signaling pathway. Casein Kinase 2 (CK2), a positive regulator of Wnt signaling, is present in beta-catenin complexes and activated in Wnt-signaling cells. We show here that CK2 enhances beta-catenin:LEF-1 transactivation in vivo and in vitro and that beta-catenin and CK2 cycle on and off the DNA in an alternating manner with the TLE1 corepressor at Wnt target genes. Interestingly, CK2 phosphorylates hLEF-1 directly and stimulates binding and transactivation of beta-catenin:LEF-1 complexes on chromatin templates in vitro. In vitro, CK2 phosphorylation of hLEF-1 strongly enhances its affinity for beta-catenin and reduces its affinity for TLE1. MALDI-TOF mass spectrometry (MS) identified two CK2 phosphorylation sites (S42, S61) within the amino terminus of hLEF-1, and mutation of these sites reduced binding to beta-catenin in vitro and transactivation in vivo. Remarkably, treatment of cells with TBB, a pharmaceutical inhibitor of CK2, blocked the recruitment and cycling of beta catenin and TLE1 at Wnt target genes in vivo. Taken together, these data indicate that CK2 is required for the assembly and cycling of Wnt-enhancer complexes in vivo. PMID- 17113389 TI - A role for kinesin-2 in COPI-dependent recycling between the ER and the Golgi complex. AB - Transport carriers operating between early compartments in the mammalian secretory pathway have to travel long distances in the cell by mostly relying on the microtubule network and its associated motor proteins. Although anterograde transport from the endoplasmic reticulum (ER) to the Golgi complex is mediated by cytoplasmic dynein, the identity of the motor(s) mediating transport in the retrograde direction is presently unclear. Some studies have suggested that the heterotrimeric kinesin-2 complex plays a role in transport between the ER and the Golgi. Here, we have examined kinesin-2 function by using an RNA-interference approach to downregulate the expression of KAP3, the nonmotor subunit of kinesin 2, in HeLa cells. KAP3 silencing results in the fragmentation of the Golgi apparatus and a change in the steady-state localization of the KDEL-receptor (KDEL-R). Using specific transport assays, we show that the rate of anterograde secretory traffic is unaffected in these cells but that KDEL-R-dependent retrograde transport is strongly abrogated. Our data strongly support a role for kinesin-2 in the KDEL-R-/COPI-dependent retrograde transport pathway from the Golgi complex to the ER. PMID- 17113390 TI - Apoptotic cells induce a phosphatidylserine-dependent homeostatic response from phagocytes. AB - Engulfment of apoptotic cells by phagocytes is important throughout development and adult life. When phagocytes engulf apoptotic cells, they increase their cellular contents including cholesterol and phospholipids, but how the phagocytes respond to this increased load is poorly understood. Here, we identify one type of a phagocyte response, wherein the recognition of apoptotic cells triggers enhanced cholesterol efflux (to apolipoprotein A-I) from macrophages. Phosphatidylserine (PS) exposed on apoptotic cells was necessary and sufficient to stimulate the efflux response. A major mechanism for this enhanced efflux by macrophages was the upregulation of the mRNA and protein for ABCA1, a membrane transporter independently linked to cholesterol efflux as well as engulfment of apoptotic cells. This increase in phagocyte ABCA1 levels required the function of nuclear receptor LXRalpha/beta, a known regulator of cholesterol homeostasis in humans and mice. Taken together, these data reveal a "homeostatic program" initiated in phagocytes that include a proximal membrane signaling event initiated by PS recognition, a downstream signaling event acting through nuclear receptors, and an effector arm involving upregulation of ABCA1, in turn promoting reverse cholesterol transport from the phagocytes. These data also have implications for macrophage handling of contents derived from apoptotic versus necrotic cells in atherosclerotic lesions. PMID- 17113391 TI - Role of CLASP2 in microtubule stabilization and the regulation of persistent motility. AB - In motile fibroblasts, stable microtubules (MTs) are oriented toward the leading edge of cells. How these polarized MT arrays are established and maintained, and the cellular processes they control, have been the subject of many investigations. Several MT "plus-end-tracking proteins," or +TIPs, have been proposed to regulate selective MT stabilization, including the CLASPs, a complex of CLIP-170, IQGAP1, activated Cdc42 or Rac1, a complex of APC, EB1, and mDia1, and the actin-MT crosslinking factor ACF7. By using mouse embryonic fibroblasts (MEFs) in a wound-healing assay, we show here that CLASP2 is required for the formation of a stable, polarized MT array but that CLIP-170 and an APC-EB1 interaction are not essential. Persistent motility is also hampered in CLASP2 deficient MEFs. We find that ACF7 regulates cortical CLASP localization in HeLa cells, indicating it acts upstream of CLASP2. Fluorescence-based approaches show that GFP-CLASP2 is immobilized in a bimodal manner in regions near cell edges. Our results suggest that the regional immobilization of CLASP2 allows MT stabilization and promotes directionally persistent motility in fibroblasts. PMID- 17113392 TI - Intracellular trafficking of interleukin-1 receptor I requires Tollip. AB - Interleukin-1 receptor (IL-1RI) is a master regulator of inflammation and innate immunity. When triggered by IL-1beta, IL-1RI aggregates with IL-1R-associated protein (IL-1RAcP) and forms a membrane proximal signalosome that potently activates downstream signaling cascades. IL-1beta also rapidly triggers endocytosis of IL-1RI. Although internalization of IL-1RI significantly impacts signaling, very little is known about trafficking of IL-1RI and therefore about precisely how endocytosis modulates the overall cellular response to IL-1beta. Upon internalization, activated receptors are often sorted through endosomes and delivered to lysosomes for degradation. This is a highly regulated process that requires ubiquitination of cargo proteins as well as protein-sorting complexes that specifically recognize ubiquitinated cargo. Here, we show that IL-1beta induces ubiquitination of IL-1RI and that via these attached ubiquitin groups, IL 1RI interacts with the ubiquitin-binding protein Tollip. By using an assay to follow trafficking of IL-1RI from the cell surface to late endosomes and lysosomes, we demonstrate that Tollip is required for sorting of IL-1RI at late endosomes. In Tollip-deficient cells and cells expressing only mutated Tollip (incapable of binding IL-1RI and ubiquitin), IL-1RI accumulates on late endosomes and is not efficiently degraded. Furthermore, we show that IL-1RI interacts with Tom1, an ubiquitin-, clathrin-, and Tollip-binding protein, and that Tom1 knockdown also results in the accumulation of IL-1RI at late endosomes. Our findings suggest that Tollip functions as an endosomal adaptor linking IL-1RI, via Tom1, to the endosomal degradation machinery. PMID- 17113393 TI - A role for Tim21 in membrane-potential-dependent preprotein sorting in mitochondria. AB - The mitochondrial inner membrane harbors complexes of the respiratory chain and translocase complexes for preproteins. The membrane potential generated by the respiratory chain is essential for ATP production by the mitochondrial ATP synthase and as a driving force for protein import. It is generally believed that the preprotein translocases just use the membrane potential without getting into physical contact with respiratory-chain complexes. Here, we show that the presequence translocase interacts with the respiratory chain. Tim21, a specific subunit of the sorting-active presequence translocase , recruits proton-pumping respiratory-chain complexes and stimulates preprotein insertion. Thus, the presequence translocase cooperates with the respiratory chain and promotes membrane-potential-dependent protein sorting into the inner mitochondrial membrane. These findings suggest a new coupling mechanism in an energy transducing membrane. PMID- 17113394 TI - Treatment of congestion in acute heart failure syndromes: importance, strategies, and challenges. Introduction. PMID- 17113395 TI - Pathophysiology of volume overload in acute heart failure syndromes. AB - The inability to effectively regulate volume status is a major consequence of acute heart failure syndromes (AHFS). A variety of pathophysiologic processes contribute to this impairment, most notably neurohormonal activation of the renin angiotensin-aldosterone system, arginine vasopressin, and the sympathetic nervous system. As a result, addressing volume overload is one of the most challenging aspects of AHFS management. Neurohormonal activation leads to substantial changes in hemodynamics and myocardial remodeling, which further contribute to the severity of heart failure (HF) disease and thereby cyclically increase the risk of further neurohormonal activation. Pulmonary capillary wedge pressure is a dependable reflection of volume status and has been used as a surrogate marker in recent studies to assess disease progression in response to innovative HF treatment strategies. Future approaches to HF treatment should focus on the more accurate assessment and management of volume status in an effort to improve patient care. PMID- 17113396 TI - The confounding issue of comorbid renal insufficiency. AB - The United States is currently beleaguered by twin epidemics, heart failure (HF) and renal insufficiency (RI). HF and RI frequently coexist in the same patient, and this conjunction, often called the "cardiorenal syndrome," has important therapeutic and prognostic implications. Approximately 60% to 80% of patients hospitalized for HF have at least stage III renal dysfunction as defined by the National Kidney Foundation (NKF), and this comorbid RI is associated with significantly increased morbidity and mortality risk. Numerous studies have demonstrated that in patients with HF, indices of renal function are the most powerful independent mortality risk predictors. Comorbid RI can result from both intrinsic renal disease and inadequate renal perfusion. Atherosclerosis, renal vascular disease, diabetes mellitus, and hypertension are significant precursors of both HF and RI. Moreover, diminished renal perfusion is frequently a consequence of the hemodynamic changes associated with HF and its treatment. Both HF and RI stimulate neurohormonal activation, increasing both preload and afterload and reducing cardiac output. Inotropic agents augment this neurohormonal activation. In addition, diuretics can produce hypovolemia and intravenous vasodilators can cause hypotension, further diminishing renal perfusion. Management of these patients requires successfully negotiating the delicate balance between adequate volume reduction and worsening renal function. Despite this, few evidence-based data are available to guide management decisions, indicating a compelling need for additional studies in this patient population. PMID- 17113397 TI - Practical applications of intravenous diuretic therapy in decompensated heart failure. AB - Intravenous (IV) loop diuretics play an important role in the treatment of decompensated heart failure (DHF). They inhibit the Na(+)-K(+)-2Cl(-) reabsorptive pump in the thick ascending limb of the loop of Henle, and the resultant natriuresis and diuresis decreases volume load, improves hemodynamics, and reduces DHF symptoms. However, loop diuretics have a short half-life and their efficacy may be limited by postdiuretic sodium rebound during the period between doses in which the tubular diuretic concentration is subtherapeutic. Moreover, they can produce electrolyte abnormalities, neurohormonal activation, intravascular volume depletion, and renal dysfunction. Several studies have reported an association between diuretic therapy and increased morbidity and mortality. In addition, many patients, especially those with more advanced forms of heart failure (HF), are resistant to standard doses of loop diuretics. These high-risk, resistant patients may benefit from pharmacologic and/or nonpharmacologic interventions to improve hemodynamic performance, treatment of renovascular disease, discontinuation of aspirin and other sodium-retaining drugs, manipulation of the route of delivery or combination of diuretic classes, or hemofiltration. Despite >50 years of use, many questions regarding the use of intravenous diuretic agents in patients with DHF are still unanswered, and there remains a compelling need for well-designed randomized, controlled clinical trials to establish appropriate treatment regimens that maximize therapeutic benefit while minimizing morbidity and mortality. PMID- 17113398 TI - Congestion in acute heart failure syndromes: an essential target of evaluation and treatment. AB - Patients with acute heart failure syndromes (AHFS) typically present with signs and symptoms of systemic and pulmonary congestion at admission. However, elevated left ventricular (LV) filling pressures (hemodynamic congestion) may be present days or weeks before systemic and pulmonary congestion develop, resulting in hospital admission. This "hemodynamic congestion," with or without clinical congestion, may have deleterious effects including subendocardial ischemia, alterations in LV geometry resulting in secondary mitral insufficiency, and impaired cardiac venous drainage from coronary veins resulting in diastolic dysfunction. It is possible that these hemodynamic abnormalities in addition to neurohormonal activation may contribute to LV remodeling and heart failure progression. Approximately 50% of patients admitted for AHFS are discharged with persistent symptoms and/or minimal or no weight loss in spite of the fact that the main reason for admission was clinical congestion. Accordingly, the assessment and management of pulmonary and systemic congestion in these patients require reevaluation. PMID- 17113399 TI - Beyond diuretics: management of volume overload in acute heart failure syndromes. AB - Diuretics are an established foundation of therapy for patients with chronic heart failure (HF) as well as for those hospitalized for treatment of acute HF syndromes. Despite the accepted use of diuretics in acute HF syndromes, treatment patterns with diuretics vary widely, and there are no data from randomized studies on the benefit of diuretics on morbidity or mortality in patients hospitalized with acute HF syndromes. Additional pharmacologic therapies that complement or replace diuretics in this setting, especially in patients with diuretic resistance, include positive inotropes, nitrovasodilators, and natriuretic peptides, but data are likewise lacking on important clinical outcomes. Ultrafiltration has also been used as a nonpharmacologic strategy to treat patients with acute HF syndromes who exhibit resistance to diuretics. Effective monitoring of volume status with newer modalities may allow more selective use of diuretics and diuretic-like modalities, but additional randomized trial data are clearly needed to establish ideal strategies to promote volume removal in acute HF syndromes. PMID- 17113400 TI - WHO 2007-12: the era of Margaret Chan. PMID- 17113401 TI - Is the sewer still the conscience of the city? PMID- 17113402 TI - A framework to slim down Europe. PMID- 17113403 TI - The ever growing story of cyclo-oxygenase inhibition. PMID- 17113404 TI - Preventing unintended pregnancy: let us count the ways. PMID- 17113405 TI - Changes in sexual behaviours to prevent HIV. PMID- 17113406 TI - The evolving doctor. PMID- 17113407 TI - Influenza vaccination for elderly people and their care workers. PMID- 17113408 TI - A gendered look at Mexico's health-sector reform. PMID- 17113409 TI - Health reform in Mexico: a work in progress. PMID- 17113410 TI - Expertise unusually redefined. PMID- 17113411 TI - Ana Langer. Interview. PMID- 17113413 TI - Intimate-partner violence. PMID- 17113414 TI - Intimate-partner violence. PMID- 17113415 TI - Intimate-partner violence. PMID- 17113416 TI - Role of drug bulletins. PMID- 17113417 TI - Intimate-partner violence. PMID- 17113419 TI - Mortality in patients with hepatitis B or hepatitis C. PMID- 17113420 TI - Chagas' disease in Mexico. PMID- 17113422 TI - Diagnosis and prevention of pertussis. PMID- 17113424 TI - Supporting physicians in low-income countries. PMID- 17113425 TI - India's efforts to boost neonatal survival. PMID- 17113426 TI - Cardiovascular outcomes with etoricoxib and diclofenac in patients with osteoarthritis and rheumatoid arthritis in the Multinational Etoricoxib and Diclofenac Arthritis Long-term (MEDAL) programme: a randomised comparison. AB - BACKGROUND: Cyclo-oxygenase-2 (COX-2) selective inhibitors have been associated with an increased risk of thrombotic cardiovascular events in placebo-controlled trials, but no clinical trial has been reported with the primary aim of assessing relative cardiovascular risk of these drugs compared with traditional non steroidal anti-inflammatory drugs (NSAIDs). The MEDAL programme was designed to provide a precise estimate of thrombotic cardiovascular events with the COX-2 selective inhibitor etoricoxib versus the traditional NSAID diclofenac. METHODS: We designed a prespecified pooled analysis of data from three trials in which patients with osteoarthritis or rheumatoid arthritis were randomly assigned to etoricoxib (60 mg or 90 mg daily) or diclofenac (150 mg daily). The primary hypothesis stated that etoricoxib is not inferior to diclofenac, defined as an upper boundary of less than 1.30 for the 95% CI of the hazard ratio for thrombotic cardiovascular events in the per-protocol analysis. Intention-to-treat analyses were also done to assess consistency of results. These trials are registered at http://www.clinicaltrials.gov with the numbers NCT00092703, NCT00092742, and NCT00250445. FINDINGS: 34 701 patients (24 913 with osteoarthritis and 9 787 with rheumatoid arthritis) were enrolled. Average treatment duration was 18 months (SD 11.8). 320 patients in the etoricoxib group and 323 in the diclofenac group had thrombotic cardiovascular events, yielding event rates of 1.24 and 1.30 per 100 patient-years and a hazard ratio of 0.95 (95% CI 0.81-1.11) for etoricoxib compared with diclofenac. Rates of upper gastrointestinal clinical events (perforation, bleeding, obstruction, ulcer) were lower with etoricoxib than with diclofenac (0.67 vs 0.97 per 100 patient-years; hazard ratio 0.69 [0.57-0.83]), but the rates of complicated upper gastrointestinal events were similar for etoricoxib (0.30) and diclofenac (0.32). INTERPRETATION: Rates of thrombotic cardiovascular events in patients with arthritis on etoricoxib are similar to those in patients on diclofenac with long term use of these drugs. PMID- 17113427 TI - Unintended pregnancy and use of emergency contraception among a large cohort of women attending for antenatal care or abortion in Scotland. AB - BACKGROUND: Unintended pregnancy is common. Although many unintended pregnancies end in induced abortion, up to a third of those proceeding to birth might be unplanned. Some of these pregnancies could be prevented by emergency contraception. We have sought to establish how many pregnancies ending in either childbirth or abortion are unintended, and what proportion of women use emergency contraception to try to prevent pregnancy. METHODS: 2908 women who attended an Edinburgh hospital for antenatal care and 907 attending for abortion fully completed a self-administered questionnaire including a validated measure of pregnancy intention and questions about emergency contraceptive use. FINDINGS: 814 (89.7%) of 907 pregnancies among women requesting abortion were unintended compared with only 250 (8.6%) among 2908 women who planned to continue pregnancy. However, only 1909 (65.6%) of continuing pregnancies were intended. The rest of the women were ambivalent about pregnancy intention. In women who continued with their pregnancies intendedness was related to age, with unintended pregnancy most probable in young women (p<0.0001). Emergency contraception was used by 113 (11.8%) of women who requested abortion but only 40 (1%) of those planning to continue pregnancy. In those whose pregnancy was continuing, the proportions reporting use of emergency contraception were higher in young women than in older women and in those who reported that their pregnancies were unintended than in those who meant to become pregnant (both p<0.0001). INTERPRETATION: Unintended pregnancy is common, even among women planning to continue pregnancy. However, EC use is low even among women with no intention of conceiving, and is thus unlikely to reduce unintended pregnancy rates. Rather, we need to find ways to improve the use of regular contraception. PMID- 17113428 TI - Sexual abstinence, contraception, and condom use by young African women: a secondary analysis of survey data. AB - BACKGROUND: Drug therapy for people with AIDS is a humanitarian priority but prevention of HIV infection remains essential. Focusing on young single African women, we aimed to assess trends in a set of behaviours-sexual abstinence, contraceptive use, and condom use-that are known to affect the rates of HIV transmission. METHODS: We did a secondary analysis of public-access data sets in 18 African countries (132,800 women), and calculated changes in a set of behavioural indicators over time. We standardised these trends from nationally representative surveys to adjust for within-country changes in age, education, and type of residential location. FINDINGS: Between about 1993 and 2001, the percentage of women reporting no sexual experience changed little. During the same period, the percentage of sexually experienced women who reported no sexual intercourse in the previous 3 months (secondary abstinence) rose significantly in seven of 18 countries and the median for all 18 countries increased from 43.8% to 49.2%. Use of condoms for pregnancy prevention rose significantly in 13 of 18 countries and the median proportion increased from 5.3% to 18.8%. The median rate of annual increase of condom use was 1.41 percentage points (95% CI 1.12-2.25). In the 13 countries with available data, condom use at most recent coitus rose from a median of 19.3% to 28.4%. Over half (58.5%) of condom users were motivated, at least in part, by a wish to avoid pregnancy. INTERPRETATION: Condom promotion campaigns in sub-Saharan Africa have affected the behaviour of young single women; the pace of change has matched the rise in contraceptive use by married couples in developing countries over recent decades. Thus continuing efforts to promote condom use with emphasis on pregnancy prevention are justified. PMID- 17113429 TI - Pneumoperitoneum following percutaneous lung biopsy. PMID- 17113430 TI - Retinitis pigmentosa. AB - Hereditary degenerations of the human retina are genetically heterogeneous, with well over 100 genes implicated so far. This Seminar focuses on the subset of diseases called retinitis pigmentosa, in which patients typically lose night vision in adolescence, side vision in young adulthood, and central vision in later life because of progressive loss of rod and cone photoreceptor cells. Measures of retinal function, such as the electroretinogram, show that photoreceptor function is diminished generally many years before symptomic night blindness, visual-field scotomas, or decreased visual acuity arise. More than 45 genes for retinitis pigmentosa have been identified. These genes account for only about 60% of all patients; the remainder have defects in as yet unidentified genes. Findings of controlled trials indicate that nutritional interventions, including vitamin A palmitate and omega-3-rich fish, slow progression of disease in many patients. Imminent treatments for retinitis pigmentosa are greatly anticipated, especially for genetically defined subsets of patients, because of newly identified genes, growing knowledge of affected biochemical pathways, and development of animal models. PMID- 17113431 TI - Family planning: the unfinished agenda. AB - Promotion of family planning in countries with high birth rates has the potential to reduce poverty and hunger and avert 32% of all maternal deaths and nearly 10% of childhood deaths. It would also contribute substantially to women's empowerment, achievement of universal primary schooling, and long-term environmental sustainability. In the past 40 years, family-planning programmes have played a major part in raising the prevalence of contraceptive practice from less than 10% to 60% and reducing fertility in developing countries from six to about three births per woman. However, in half the 75 larger low-income and lower middle income countries (mainly in Africa), contraceptive practice remains low and fertility, population growth, and unmet need for family planning are high. The cross-cutting contribution to the achievement of the Millennium Development Goals makes greater investment in family planning in these countries compelling. Despite the size of this unfinished agenda, international funding and promotion of family planning has waned in the past decade. A revitalisation of the agenda is urgently needed. Historically, the USA has taken the lead but other governments or agencies are now needed as champions. Based on the sizeable experience of past decades, the key features of effective programmes are clearly established. Most governments of poor countries already have appropriate population and family-planning policies but are receiving too little international encouragement and funding to implement them with vigour. What is currently missing is political willingness to incorporate family planning into the development arena. PMID- 17113432 TI - Evidence is good for your health system: policy reform to remedy catastrophic and impoverishing health spending in Mexico. AB - Absence of financial protection in health is a recently diagnosed "disease" of health systems. The most obvious symptom is that families face economic ruin and poverty as a consequence of financing their health care. Mexico was one of the first countries to diagnose the problem, attribute it to lack of financial protection, and propose systemic therapy through health reform. In this article we assess how Mexico turned evidence on catastrophic and impoverishing health spending into a catalyst for institutional renovation through the reform that created Seguro Popular (Popular Health Insurance). We present 15-year trends on the evolution of catastrophic and impoverishing health spending, including evidence on how the situation is improving. The results of the Mexican experience suggest an important role for the organisation and financing of the health system in reducing impoverishment and protecting households during periods of individual and collective financial crisis. PMID- 17113433 TI - Caecal cancer associated with longstanding Crohn's disease. PMID- 17113436 TI - The use of internal maxillary distraction for maxillary hypoplasia: a preliminary report. AB - PURPOSE: Distraction osteogenesis is a useful alternative to advance the maxilla in complicated cases of maxillary hypoplasia. The purpose of this article is to review the workup, experience, and preliminary results with the use of internal distraction osteogenesis for maxillary hypoplasia at one teaching institution. PATIENTS AND MATERIALS: Over a 5-year period, more than 300 patients with craniofacial and dentofacial defects have undergone oral and maxillofacial surgery at our center to correct their skeletal discrepancies. Of these, 10 have had maxillary distraction osteogenesis done with internal distractors. Follow-up of 6 months or more was available for 8 patients. Stereolithographic models were used to bend distractors prior to surgery in 6 patients. RESULTS: Latency prior to the start of distraction was 3 to 7 days and varied with the age of the patient. Distraction occurred at approximately 1 mm per day with an average distraction length of 8.5 mm (range, 6-10 mm). Excellent occlusal results were obtained in 5 patients. Major complications including nonunion and failure to achieve acceptable occlusal results were observed in 3 patients. Minor complications including pain and loosening of the distracter devices were observed in 2 patients, but did not appear to affect the esthetic and functional results. CONCLUSIONS: Distraction osteogenesis is a useful alternative to traditional orthognathic surgery to treat maxillary hypoplasia. Internal distractions are attractive to patients, but are more difficult to place and can cause discomfort to patients when trying to achieve an ideal primary vector of distraction. Stereolithographic models can help with placement of the device. Changes in design of distractors may help with patient discomfort. PMID- 17113437 TI - Assessment of oral health-related quality of life before and after third molar surgery. AB - PURPOSE: This study was designed to further our understanding of recovery after third molar surgery by using 2 instruments to measure quality of life outcomes, the more global Oral Health Impact Profile (OHIP-14), and the condition-specific Health-Related Quality of Life (HRQOL) instrument. PATIENTS AND METHODS: Clinical and quality of life data pre- and postsurgery from 63 patients with all 4 third molars below the occlusal plane, treated with topical minocycline during third molar surgery to reduce the incidence of delayed clinical healing, were available for analyses. Each patient was given 2 questionnaires to complete; the more global OHIP-14 and the HRQOL instrument designed to assess recovery after third molar surgery. Prevalence, Extent, and Severity of the OHIP-14 scores were calculated presurgery and for postsurgery days (PSD) 1, 7, 14. The percentage of patients reporting clinically relevant responses detrimental to quality of life from the condition-specific HRQOL instrument were reported for the same time frame. RESULTS: Study patients were most likely female, less than 25 years old, and Caucasian. Most (72%) had bone removed from both lower third molars. Median surgery time was 27 minutes (interquartile range [IQ], 20, 40 minutes.). Median surgeons' estimate of overall difficulty was 14 of a possible 28 (IQ 10, 18), and the median degree of difficulty for lower third molars was 8 of 14 (IQ 6, 10). Few patients (only 10%) had delayed clinical healing. Prevalence for all OHIP-14 items, percent of patients reporting items "fairly often" or "very often," were increased from presurgery on PSD 1 and then decreased on PSD 7 and PSD 14. OHIP 14 Severity scores, the sum of OHIP-14 responses, followed the same pattern as the Prevalence scores. OHIP-14 Severity scores on PSD 1 were 27 (IQ 16, 34), decreasing to 8 (IQ 3, 13) by PSD 7, and 1 (IQ 0, 5) by PSD 14. Recovery for outcomes addressed by both instruments followed a similar pattern and time course. However, each instrument also assessed distinctly different outcomes, adding information that could not be obtained by 1 instrument alone. CONCLUSION: Complementary instruments to measure quality of life outcomes provide a broader understanding of recovery after third molar surgery. PMID- 17113438 TI - A retrospective analysis of oral and maxillofacial injuries in motor vehicle accidents. AB - PURPOSE: To clarify the relations of the severity of oral and maxillofacial injuries from traffic accidents (TAs) to seating position and the use of restraint systems. PATIENTS AND METHODS: Hospital records were reviewed for all patients who had sustained oral or maxillofacial injuries in TAs and then were admitted to the Department of Oral and Maxillofacial Surgery, Dokkyo University School of Medicine (Tochigi, Japan), from 1994 through 2003. RESULTS: A total of 201 patients, with a mean injury severity score (ISS) of 7.9 +/- 7.6, were included in this study. Although patients with any oral or maxillofacial injury with an Abbreviated Injury Scale (AIS) score of 2 or more had extremely low ISSs, hospitalization was relatively long. The ISS and AIS score of the head or neck were significantly higher in unrestrained drivers (12.4 +/- 11.2, 1.2 +/- 1.4, respectively) than in restrained drivers (6.5 +/- 4.6, 0.4 +/- 1.0, respectively). However, AIS scores of the face were similar in unrestrained drivers (1.9 +/- 0.7) and restrained drivers (1.7 +/- 0.5). Furthermore, the incidence of maxillofacial fractures did not differ between the 2 groups. CONCLUSION: Because wearing seat belts cannot prevent all oral and maxillofacial injuries in motor vehicle occupants, both physicians and engineers must pay greater attention to the mechanisms of oral and maxillofacial injuries in TAs. PMID- 17113439 TI - Speech outcome after cranial-based pharyngeal flap in children born with total cleft, cleft palate, or primary velopharyngeal insufficiency. AB - PURPOSE: The aim of this study was to compare the effect of a cranial-based pharyngeal flap on the speech of children born with a unilateral cleft lip and palate (UCLP), bilateral cleft lip and palate (BCLP), cleft palate (CP), or primary velopharyngeal insufficiency (VPI) without cleft. PATIENTS AND METHODS: A total of 234 children born with clefts and 22 children born with primary VPI were evaluated. Children with associated abnormalities were excluded from this study. The Borel-Maisonny classification system was used to evaluate the velar insufficiency. The cranial-based pharyngeal flap was performed using the Sanvenero-Rosselli technique. RESULTS: Between 1984 and 2001, 74 children underwent pharyngeal flap for VPI. The mean follow-up period was 7 years. Borel Maisonny scores after pharyngeal flap surgery were as follows: children with UCLP (n = 22), 59.1% type 1, 36.4% type 1/2, and 4.5% type 2; children with BCLP (n = 18), 44.4% type 1, 27.8% type 1/2, 16.7% type 2, and 11.1% type 2/3; children with CP (n = 17), 64.7% type 1, 23.5% type 1/2, and 11.8% type 2; children with primary VPI (n = 17), 29.4% type 1, 29.4% type 1/2, 29.4% type 2/3, and 11.8% type 3. There were significant differences in outcome among the 4 groups (P = .029; Fisher exact test). CONCLUSIONS: The positive effect on speech of a cranial based pharyngeal flap is greater in children born with a UCLP or CP than in those born with a BCLP. In children born with primary VPI, this operation has only a slightly positive effect on speech that shows compensatory misarticulations; in such cases, alternative surgical choices or secondary procedures may be indicated. This information should be clearly conveyed to the parents in presurgical consultation so that they know what to expect from the procedure and postoperative adjuvant therapy. PMID- 17113440 TI - The early psychological adjustment of cleft patients after maxillary distraction osteogenesis and conventional orthognathic surgery: a preliminary study. AB - PURPOSE: To compare the early psychological changes of cleft lip and palate (CLP) and noncleft patients after maxillofacial corrective surgery, including maxillary distraction osteogenesis and conventional orthognathic surgery. MATERIALS AND METHODS: Nine CLP patients were compared with a group of 9 non-CLP patients having similar dentofacial deformities in a prospective longitudinal cohort study. Five of the CLP patients underwent maxillary distraction osteogenesis and 4 underwent conventional orthognathic surgery. A control group of 9 noncleft patients received conventional orthognathic surgery. All patients completed a set of questionnaires to enable their psychological profile to be assessed. The data were collected immediately before surgery (T1), and at 3 weeks (T2) and 12 weeks (T3) after surgery. RESULTS: The CLP patients treated with distraction osteogenesis were happier, but had a higher level of social anxiety and distress than the CLP patients receiving conventional orthognathic surgery. On the other hand, the CLP patients overall were happier, with lower social anxiety and distress, than the noncleft control group. The CLP patients showed a higher level of parental self-esteem than the noncleft patients. CONCLUSION: This preliminary study shows that CLP patients were generally happier, and had a higher level of parental support, than normal patients suffering from dentofacial deformities. Maxillary distraction osteogenesis seemed to induce a higher level of anxiety and distress in CLP patients than conventional orthognathic surgery in both cleft and noncleft patients. PMID- 17113441 TI - Qualitative descriptors used by patients following orthognathic surgery to portray altered sensation. AB - PURPOSE: Following orthognathic surgery, patients use qualitatively different words to describe the altered sensation on their face that results from tissue inflammation and nerve injury. These words indicate normal, hypoesthetic, paresthetic, and dysesthetic sensations, and reflect the intrusiveness of the alteration. Our intent was to study the words chosen by patients from a standardized list to characterize sensory recovery during the first 6 months after surgery and to examine whether patients who underwent different surgical procedures tended to choose different sets of words. PATIENTS AND METHODS: Patients' selections from a list of 27 words that described their assessment of spontaneous and evoked facial sensations were obtained before surgery and at 1 week, 1 month, 3 months, and 6 months after surgery. Data were obtained from 146 patients enrolled in a randomized controlled clinical trial designed to evaluate the potential of sensory retraining in the rehabilitation of patients who experience impairment in sensory function after nerve injury. Mantel Haenszel general correlation and row mean score statistics were used to assess the association between time and word choice and to compare the word choice categories of 4 surgical groups: bilateral sagittal split osteotomy (BSSO) only, with or without genioplasty; BSSO + Le Fort I, with or without genioplasty. RESULTS: In general, the number of words selected to describe the alteration in sensation decreased over time, as did the intrusiveness of the category of words chosen. However, the intrusiveness remained the same or worsened from 1 week to 6 months for 32% of patients. With increased time after surgery, the percentage of patients who reported altered evoked sensations exceeded the percentage who reported spontaneous sensations. For example, at 6 months the altered sensation of 66% of the patients was classified in the paresthesia and dysesthesia categories by the evoked assessment of sensation; whereas, that of only 47% of the patients were classified as such by the spontaneous assessment. The addition of Le Fort I to BSSO did not affect the way patients reported altered sensation on their lower face. Hypoesthesia and paresthesia, but not dysesthesia, were less of a problem on the midface than on the lower face after BSSO + Le Fort I. Patients who had genioplasty more frequently chose descriptors for the lower face that reflected soft tissue trauma and inflammation ("swollen," "tender," and "burning") than patients without genioplasty; however, this difference decreased with time after surgery. CONCLUSION: The current findings indicate that patients' selection of words differentiates individuals who experienced only a simple loss in sensation (ie, present negative symptoms), those who experienced active sensations that are not normally present (ie, present positive symptoms), and those whose active sensations are additionally uncomfortable or painful. It is possible that continued study of the latter group of patients will reveal patterns of word usage that predict poor long-term recovery and disabling sensory disorders. PMID- 17113442 TI - The effect of a single dose prednisolone with and without diclofenac on pain, trismus, and swelling after removal of mandibular third molars. AB - PURPOSE: The purpose of this study was to evaluate the effect of a single intramuscular dose of prednisolone and the prednisolone-diclofenac combination on postoperative pain, trismus, and edema after the removal of third molars. PATIENTS AND METHODS: Forty-five patients who were to undergo surgical removal of lower third molars were studied. Patients were divided into 3 groups. In the first group, each patient was given 25 mg prednisolone intramuscularly immediately after surgery. In the second group, each patient was given 25 mg prednisolone and diclofenac intramuscularly immediately after surgery, and in the third group, each patient was given sterile saline solution as control group. Postoperative pain was evaluated by visual analogue scale on the day of surgery. Facial swelling and trismus were evaluated on postoperative days 2 and 7. ANOVA was used to analyze these data. RESULTS: Statistical analysis of the data indicated the prednisolone-diclofenac combination suppressed pain intensity in comparison with control (P < .05) at the 6-hour observation. Both the prednisolone and prednisolone-diclofenac combinations suppressed pain at the seventh postoperative hour in comparison with the control (P < .05). The prednisolone-diclofenac combination group also had a smaller loss of opening at postoperative days 2 and 7 in comparison with both the prednisolone and control groups (P < .05). Postoperative swelling was less in both the prednisolone and prednisolone-diclofenac combination groups, as compared with the control group (P < .05) at postoperative day 2. The prednisolone-diclofenac combination group also had a smaller swelling at postoperative day 7 in comparison with both the prednisolone and control groups (P < .05). CONCLUSION: It was determined that the combination of a single dose of prednisolone and diclofenac is well-suited to the treatment of postoperative pain, trismus, and swelling after dental surgical procedures and should be used when extensive postoperative swelling of soft tissue is anticipated. PMID- 17113443 TI - Outcome assessment of inferior alveolar nerve microsurgery: a retrospective review. AB - PURPOSE: This retrospective study was performed to assess the clinical outcome of patients who have undergone trigeminal nerve microsurgical repair of the inferior alveolar nerve. MATERIALS AND METHODS: This study includes all patients who underwent microsurgical repair of the inferior alveolar nerve at the University of Medicine and Dentistry of New Jersey from July 1, 1998 through June 30, 2003. Each patient underwent a thorough evaluation of sensory nerve function that included the type of injury, date of injury, and neurosensory testing. The evaluation was performed pre- and postoperatively to assess sensory improvement. Through chart review and quantitative statistical analysis, the outcome of inferior alveolar nerve microsurgical repair was assessed to ascertain which sensory variables were statistically significant in showing improvement from microsurgical procedures. RESULTS: Thirty-two patients underwent microsurgical repair of their inferior alveolar nerve injury by the same surgeon. The average period of time from initial nerve injury until primary surgical repair was 6.6 months. Four patients did not follow-up postoperatively and were excluded from the final data. Of the remaining 28 patients, mean follow-up period was 9.5 months. It was determined that 26 patients (92.9%) had statistically significant neurosensory improvement, with 14 reporting (50%) significant improvement, 12 patients (42.9%) with slight improvement, and 2 patients (7.1%) demonstrating no improvement. No statistical evidence was found to support that a decrease in time from injury to surgery had improved results in this limited patient population. CONCLUSION: Microsurgical repair provides an improvement in neurosensory function in patients that present with an inferior alveolar nerve injury. PMID- 17113444 TI - Open reduction and rigid internal fixation of mandibular condylar fractures by an intraoral approach: a long-term follow-up study of 15 patients. AB - PURPOSE: To evaluate the long-term results obtained with open reduction and rigid internal fixation of mandibular condylar fractures by an intraoral approach. PATIENTS AND METHODS: Fifteen patients with 24 mandibular condylar fractures were retrospectively examined with an average follow-up of 23 months (range, 6-63 months). Clinical and radiographic examination was conducted according to the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD), including an evaluation of maximum voluntary bite force measurements and facial nerve function. Statistical analysis was performed on maximum voluntary bite force measurements and maximum pressure pain threshold. RESULTS: Two patients fulfilled the criteria for a RDC/TMD diagnosis. Myofacial pain (group I) and bilateral arthralgia (group III), combined with a moderate nonspecific physical symptom score, was diagnosed in 1 patient and 1 patient received a diagnosis of disc displacement with reduction (group II). Satisfying radiographic fracture healing was seen in 12 joints. However, miniplate fracture occurred in 3 patients and severe bone resorption of the condylar head was seen in one patient. Minor adjustment of the postoperative occlusion was necessary in 6 patients. No significant difference between maximum voluntary isometric bite force measurements or maximum pressure pain threshold was found between the fracture side and the opposite side in unilateral cases or between the operated and nonoperated side in bilateral cases. None of the patients showed facial nerve injury or visible facial scars. CONCLUSION: Within the limitations of a retrospective study, the present study emphasized that optimal management of dislocated bilateral condylar injuries combined with other fractures of the facial skeleton constitute a challenging issue in maxillofacial trauma. Moreover, open reduction and rigid internal fixation of mandibular condylar fractures by an intraoral approach is a technically demanding surgical procedure associated with a high risk of postoperative complications in these injuries. PMID- 17113445 TI - Cost-effectiveness analysis for computer-aided surgical simulation in complex cranio-maxillofacial surgery. AB - PURPOSE: The purpose of this study is to assess the costs and benefits of computer-aided surgical simulation (CASS) and to compare it with the current surgical planning methods for complex cranio-maxillofacial (CMF) surgery. MATERIALS AND METHODS: The comparison of methods applies to all CMF surgeries where the patient's condition is severe enough to undergo a computed tomography scan and a stereolithographic model is necessary for the surgical planning process. The costs for each method can be divided into time and other costs. The time was estimated based on the authors' experience as well as on a survey of a small group of 6 experienced CMF surgeons in the United States. The other costs were estimated based on the authors' experience. RESULTS: CASS has lower costs in terms of surgeon time, patient time, and material costs. Specifically, total surgeon hours spent in planning are 5.25 hours compared with 9.75 for current standard methods. Material and scanning costs are Dollars 1,900 for CASS compared with about Dollars 3,510 for standard methods. Patient time for planning is reduced from 4.75 hours to 2.25 hours with CASS. The reduction in both time and other costs remains when the fixed fee costs of CASS are added to the variable costs. Amortized across the 600 patients per year (1,800 for the assumed 3-year life of the training and software), this adds only a few dollars and a fraction of an hour per surgery. Even in the case of a small clinic when the cost is amortized for 6 patients per year (18 patients for the assumed 3-year life of the training and software), the per surgery costs (9.65 hours and Dollars 2,456) will still favor CASS. CONCLUSION: Any great new design should consist of at least 2 of the 3 following features: faster, cheaper, and better outcome. This analysis demonstrates that CASS is faster and less costly than the current standard planning methods for complex CMF surgery. Previous studies have also shown that CASS results in better surgical outcomes. Thus, in all regards, CASS appears to be at least as good as the current methods of surgical planning. PMID- 17113446 TI - Repair of nasal complex fractures and the need for secondary septo-rhinoplasty. AB - PURPOSE: This study was undertaken to evaluate the efficacy of closed reduction of nasal fractures and determine the incidence of the need for post-traumatic septo-rhinoplasty in the management of residual nasal deformities. PATIENTS AND METHODS: Medical records of all patients with nasal fractures evaluated and treated by the Division of Oral and Maxillofacial Surgery at the University of Florida Health Science Center, Jacksonville, FL between January 2001 and October 2004 were retrospectively evaluated. Out of a total of 344 patients, 50 patients met the inclusion criteria. Forty-four patients (group A) underwent closed reduction of nasal bones and septum along with a septoplasty if needed within 2 weeks of initial injury. Six patients (group B) could not tolerate any surgical intervention because of multisystem injury or comorbidities. All 50 patients were then followed up in the Division of Oral and Maxillofacial Surgery to determine overall efficacy of the initial treatment modality, as well as the need for secondary post-traumatic septo-rhinoplasty. Group A was then further subdivided into groups A1 and A2. Group A1 consisted of patients who underwent a closed reduction of their nasal complex fracture without a residual deformity or the need for a secondary post-traumatic septo-rhinoplasty. Group A2 consisted of patients who underwent a closed reduction of their nasal complex fracture and developed a secondary nasal deformity significant enough to require a septo rhinoplasty. RESULTS: The follow-up period ranged from 1 week to 12 months. Nine patients in group A were lost to follow-up. Patients in group A1 (31 patients) were pleased with their results and did not require a secondary surgery. Four patients developed a post-traumatic nasal deformity requiring a post-traumatic septo-rhinoplasty (group A2). All patients in group B required post-traumatic septo-rhinoplasty. CONCLUSION: Closed reduction of nasal fractures appears to be an effective method of treatment as long as careful attention is paid to the key regions in the nasal complex, including the septum at the initial time of treatment. Ideal results are obtained when surgery is performed within 2 weeks of initial injury. Factors such as timing of surgery, the status of the nasal septum, delay in treatment, and other associated injuries may influence the overall result. PMID- 17113447 TI - Etiology of lingual nerve injuries in the third molar region: a cadaver and histologic study. AB - PURPOSE: It has been suggested that different etiologies of lingual nerve damage in the third molar area will produce a different clinical and histologic appearance in the nerve. If the clinical and histologic pictures were different, it could result in different treatments being recommended. MATERIALS AND METHODS: Eight preserved cadavers (16 lingual nerves) were used for this study. As far as possible, the nerves were left in situ and damaged in a way that could be envisaged during third molar surgery. In each case, the damaged sections of nerve were photographed, resected, embedded in paraffin wax, sectioned in 5 mum sections, stained with hematoxylin-eosin, and examined histologically. RESULTS: The scalpel clinically produced a clean wound with sharply defined edges; this was confirmed histologically with minimal disruption to the fascicles. The 702 fissure bur produced a ragged stretch-type injury clinically, and histologically this was confirmed with an irregular-edged border to the lesion and stretching and internal damage to the fascicles immediately adjacent to the wound. The crush injury clinically caused considerable apparent damage to the nerve, which was confirmed histologically with crushing and disruption of the fascicles and reduction to approximately 25% of their preinjury thickness. The stretch injury clinically showed no damage, but histologically showed irregular internal disruption of the fascicles over the whole area subject to stretching movements. CONCLUSION: It does appear that different modalities in nerve injury produce a different type of injury both clinically and histologically. This information has implications for both natural clinical recovery and the indications for surgical intervention. Clinical recovery may occur best with close approximation of a sharp scalpel-type wound or excision of a crushed area of nerve with reapproximation of the nerve endings, but a ragged wound caused by a fissure bur may require excision back to healthy nerve with subsequent reapproximation, whereas with the stretching injury it may be difficult to ascertain the edges and limits of the wound, and difficult to repair, and it may be most appropriate to rely on a natural healing process for the best results. PMID- 17113448 TI - Fatal rhino-orbito-cerebral zygomycosis caused by Apophysomyces elegans in a healthy patient. PMID- 17113449 TI - Training for oral and maxillofacial surgery, academic oral surgery, and surgical dentistry in the United Kingdom. PMID- 17113450 TI - The case against a 2-tiered training system. PMID- 17113451 TI - Removal of asymptomatic third molars: a supporting view. PMID- 17113452 TI - Removal of asymptomatic third molars: an opposing view. PMID- 17113453 TI - Traumatic changes of the inferior alveolar nerve and Gasserian ganglion after removal of a mandibular third molar: report of a case. PMID- 17113454 TI - Trans-geniohyoid dermoid cyst: considerations on a combined oral and dermal surgical approach and on histogenesis. PMID- 17113455 TI - Recurrent superficial mucoceles associated with lichenoid disorders. PMID- 17113456 TI - Removal of a large odontoma by sagittal split osteotomy. PMID- 17113458 TI - A changing paradigm of glioma biology. AB - The past 30 years have witnessed a major paradigm shift in brain tumor research with the development of a wide variety of molecular biology techniques. These methods have permitted a better understanding of the pathogenesis of gliomas including the finding of neural stem cells that contribute to the establishment and continuous population of brain tumors. Molecular biology has contributed to our understanding of prognosis in these tumors with findings of genetic correlations to patient age, response to treatment, and outcome. Gene therapy has been made possible by molecular techniques that contribute to new treatment options. Nevertheless, if these advances are to make substantial clinical improvements, attention must be paid to issues of tumor evolution, local versus general pathogenesis, tumor heterogeneity, both general and regional, and the development of resistance to treatment. Appropriate clinical trials will be needed to test these new findings. PMID- 17113459 TI - Glioma oncogenesis and animal models of glioma formation. AB - Recent advances in animal models have improved our understanding of the pathway abnormalities driving glioma growth. This article reviews key molecular abnormalities that have been modeled in mice, and describes major tumor modeling techniques along with examples of astrocytoma and oligodendroglioma models. Animal models are important not only for the testing of novel therapeutics but also as a means to understand the molecular explanations for treatment success and failure in humans. PMID- 17113460 TI - Multi-modality molecular imaging of tumors. AB - Noninvasive in vivo molecular-genetic imaging uses nuclear, magnetic resonance, and optical imaging techniques. Described and discussed are "direct" imaging of specific molecules and pathway activity, "indirect" reporter gene imaging, and "bio-marker" or "surrogate" imaging. Applications of PET- and optical-based reporter imaging are demonstrated, including imaging of oncogenesis in genetic mouse models, endogenous molecular-genetic-biological properties, and response to therapy in animal models of human disease. Molecular imaging studies complement established ex vivo molecular-biological assays that require tissue sampling by providing a spatial as well as temporal dimension to our understanding of oncogenesis, and the progression and treatment of cancer. Molecular imaging studies being performed in experimental animals will be translated to animals in the near future. PMID- 17113461 TI - The changing management of low-grade astrocytomas and oligodendrogliomas. AB - Low-grade gliomas are uncommon primary brain tumors that preferentially affect young to middle-aged adults. Although they are indolent tumors, low-grade gliomas cause considerable and progressive morbidity and are ultimately fatal. Surgery and radiotherapy are the primary therapeutic options for patients with these diseases. Chemotherapy is playing a larger role in the management of patients with low-grade gliomas. Patients with oligodendrogliomas or other low-grade gliomas that harbor a distinct genetic derangement characterized by allelic loss of chromosomes 1p and 19q appear to have a superior prognosis that is due in part to a more predictable and durable response to treatment. For this subset of patients with low-grade gliomas, treatment with initial chemotherapy and deferred radiotherapy is an increasingly attractive therapeutic approach. PMID- 17113462 TI - An update on primary central nervous system lymphoma. AB - The most important recent advance in treatment of primary central nervous system lymphoma has been the introduction of high-dose methotrexate-based chemotherapy. Convincing data demonstrate that the regimens of such chemotherapy improve survival compared with historical controls treated with radiotherapy alone. However, the optical treatment approach is still unclear and therapy can be associated with long-term neurotoxicity. Current research focuses on maximizing survival while minimizing neurologic sequelae. PMID- 17113463 TI - Chemotherapy and the treatment of brain metastases. AB - Brain metastases have traditionally been treated with a surgical or radiotherapeutic approach. Chemotherapy is used occasionally as salvage therapy. The blood-brain barrier excludes most chemotherapeutic agents, rendering many systemic options ineffective within the CNS. Intrathecal chemotherapies do not penetrate into brain tissue or bulky parenchymal tumors, so are ineffective in treatment of brain metastases. However, some patients with brain metastases benefit from chemotherapy, and temozolomide or targeted therapies like gefitinib have demonstrated activity. A better understanding of the biological behavior of brain metastases may lead to development of effective treatments for this common complication of systemic cancer. The review discusses the biology of brain metastases and provides an update on current chemotherapeutic strategies. PMID- 17113464 TI - Clinical approach to metastatic epidural spinal cord compression. AB - Metastatic epidural spinal cord compression (MESCC) is a devastating complication of cancer that occurs when cancer metastasizes to the spine and then secondarily compresses the spinal cord. It is a relatively common complication of cancer and. in the United States, more than 20,000 cases of MESCC are diagnosed annually. If left untreated, virtually 100% of these patients would become paraplegic; therefore, it is considered a true medical emergency and immediate intervention is required. Even with aggressive therapy, results can often be unsatisfactory. Although most patients with MESCC have limited survival, up to one third will survive beyond one year. Thus. it is essential to consider aggressive therapy to preserve or improve the quality of life and prevent paraplegia. PMID- 17113465 TI - Surgical approach to epidural spinal cord compression. AB - NOMS provides a framework to make decisions regarding surgery or radiation in the face of changing technology. NOMS reflects the most important decision points including neurologic, oncologic, mechanical stability, and systemic disease. Currently, patients who have high-grade epidural spinal cord compression (N) from radioresistant disease (O) or demonstrate mechanical instability (M) are offered surgery if they can tolerate it from a systemic (S) standpoint. Patients with radiosensitive tumors (O) are offered external beam radiation regardless of the degree of spinal cord compression (N). Patients with radioresistant tumors (O) who do not have significant spinal cord compression (N) are now offered IGIMRT as the best chance of controlling local tumor and avoiding an operation. PMID- 17113467 TI - Supportive care of brain tumor patients. AB - The supportive care of patients who have brain tumors consists mainly of the treatment of brain edema, seizures, venous thromboembolism, and cognitive dysfunction. Each of these complications may occur in patients who have primary or metastatic brain tumors. The development of any of these complications significantly increases the morbidity and mortality associated with brain tumors. Effective treatment is usually possible, however, and can result in an improved quality of life for these patients. PMID- 17113466 TI - Clinical and immunological diversity of limbic encephalitis: a model for paraneoplastic neurologic disorders. AB - The most important contribution to the understanding and management of paraneoplastic neurologic disorders (PND) is the discovery that many of these diseases are immune mediated. It is believed that cytotoxic T-cell responses and antibodies that target neuronal proteins usually expressed by the underlying tumor cause the neurologic symptoms. The detection of these antibodies has provided diagnostic tests that allow recognition of the disorder as paraneoplastic and direct the search of the tumor to selected organs. This article summarizes the authors' findings of limbic encephalitis and postulates that a similar approach can be used for syndromes involving other areas of the nervous system. PMID- 17113468 TI - Cognitive functions in brain tumor patients. AB - As effective treatment interventions have increased survival rates, there has been greater awareness that many brain tumor patients experience cognitive dysfunction despite adequate disease control. Cognitive difficulties often have an impact on quality of life and interfere with the patient's ability to function at premorbid levels; however, the incidence of cognitive dysfunction in brain tumor patients is unknown, because it has not been investigated systematically. Future prospective clinical trials in neuro-oncology should include cognitive outcome measures to increase understanding of the contribution of the tumor and the delayed effects of treatment to cognitive dysfunction. PMID- 17113469 TI - Central nervous system toxicity from cancer therapies. AB - The central nervous system (CNS) is an organ with a unique profile of vulnerability to antineoplastic treatments. In many cases, CNS neurotoxicity is the dose-limiting side effect of treatment for systemic and CNS neoplasms. Novel methods of delivering radiation and chemotherapy agents have led to recognition of new forms of CNS neurotoxicity. In this article, the authors review the most important CNS toxicities of cancer treatment. PMID- 17113470 TI - The ultrastructure and reproduction of Amphiamblys capitellides (Microspora, Metchnikovellidae), a parasite of the gregarine Ancora sagittata (Apicomplexa, Lecudinidae), with redescription of the species and comments on the taxonomy. AB - The ultrastructural cytology and reproduction of the hyperparasitic microsporidium Amphiamblys capitellides (Caullery and Mesnil, 1897) is described. Merogonial reproduction was not observed. The sporogony comprises two sequences: a sac-bound sporogony in close contact with the cytoplasm of the host and a free sporogony in parasitophorous vacuoles. The free sporogony, which probably precedes the sac-bound, yields a small number of rounded spores. The sac-bound sporogony is polysporoblastic, generating two rows of elongated spores. All stages have isolated nuclei. Both spore types have an extrusion apparatus of the metchnikovellidean type, with a polar sac devoid of anchoring disc, a polar filament with one manubroid and one bulbous part, and a posterior semicircular membrane fold enclosing rounded or tubular structures. Hosts are gregarines of the species Ancora sagittata living in the intestine of polychaetes of the genus Capitella, probably the species Capitella giardi. The cytology, life cycle and classification are discussed. The species is redescribed and the diagnosis of the genus Amphiamblys Caullery and Mesnil, 1914 is emended. PMID- 17113471 TI - Rigidothrix goiseri nov. gen., nov. spec. (Rigidotrichidae nov. fam.), a new "flagship" ciliate from the Niger floodplain breaks the flexibility-dogma in the classification of stichotrichine spirotrichs (Ciliophora, Spirotrichea). AB - Rigidothrix goiseri nov. gen., nov. spec. was discovered in soil from the Niger floodplain near to the town of Timbuktu, Republic of Mali. Its morphology, ontogenesis, and 18S rDNA gene sequence were studied with standard methods. Rigidothrix goiseri is very conspicuous in vivo because of its average size of 230x70 microm and a distinct tail. Further main characteristics include the rigid body; the conspicuous, stylonychid frontal area; the undulating membranes in Oxytricha pattern; a mighty adoral zone of membranelles not reorganized during ontogenesis; distinct midventral rows of which those of the opisthe develop right of the parental ones; and eight dorsal kineties, of which three develop intrakinetally and five are generated dorsomarginally. Rigidothrix goiseri possesses main characteristics of oxytrichine, stylonychine, and urostyline stichotrichs, specifically, it is the first rigid stichotrich with midventral rows, and thus it breaks the flexibility dogma. Distinct similarities with the genus Uroleptus and the CEUU hypothesis suggest that R. goiseri is more closely related to the oxytrichine than urostyline stichotrichs, in spite of the conspicuous midventral rows. This is also supported by the sequence data which show that R. goiseri is almost equally similar to both Oxytricha granulifera (95.88%) and Uroleptus gallina (94.93%), but fairly different from Urostyla grandis (92.7%). The main morphological peculiarities of R. goiseri show that it represents a new genus which should be classified into a new family, the Rigidotrichidae, together with the genera Uroleptus, Territricha, and Afrophrya nov. gen., the latter comprising Rigidothrix-like stichotrichs with cyrtohymenid oral apparatus. The conspicuous size and shape make R. goiseri a biogeographic flagship likely confined to Africa. PMID- 17113472 TI - Pseudovorticella zhengae n. sp., P. difficilis (Kahl, 1933) Jankowski, 1976, and P. punctata (Dons, 1918) Warren, 1987, three marine peritrichous ciliates from north China. AB - The morphology, infraciliature and silverline system of three marine peritrichous ciliates, Pseudovorticella zhengae n. sp., P. difficilis (Kahl, 1933) Jankowski, 1976 and P. punctata (Dons, 1918) Warren, 1987, were investigated by observation in vivo and silver-staining methods. The new species is characterized by its body shape, the appearance of the peristomial lip, the number of transverse silverlines, and the configuration of infundibular polykinety 3; P. difficilis by its body shape, the configuration of infundibular polykinety 3, and unusual annular swellings on the stalk; and P. punctata by the appearance of the pellicle, the configuration of infundibular polykinety 3, and the number of transverse silverlines. PMID- 17113473 TI - Taxonomic characterization of two marine peritrichous ciliates, Epicarchesium corlissi n. sp. and Pseudovorticella jiangi n. sp. (Ciliophora: Peritrichia), from northern China. AB - Two new marine peritrich ciliates, Epicarchesium corlissi n. sp. and Pseudovorticella jiangi n. sp., were discovered in mariculture waters on the coast of northern China near Qingdao. Their morphology, infraciliature and silverline system were investigated based on both living and silver-impregnated specimens. E. corlissi is characterized as follows: marine Epicarchesium with dichotomously branched stalk; zooids elongate, approximately 60-70 x 25-35 microm in vivo; peristomial collar double-folded; macronucleus J-shaped; single, small contractile vacuole ventrally positioned; more than 60 striations between peristome and aboral trochal band, 13-18 from aboral trochal band to scopula; abstomal end of row 1 of infundibular polykinety 3 terminating at same level as rows 2 and 3 of infundibular polykinety 3; rows 2 and 3 of infundibular polykinety 3 much longer than row 1 and converging adstomally with infundibular polykinety 1. The new species P. jiangi is diagnosed as follows: marine Pseudovorticella; zooid inverted bell-shaped, approximately 80x60 microm in vivo and with a broad, flat, thin peristomial collar that measures approximately 90 microm across; pellicle with transparent cortical vesicles; macronucleus J shaped; number of silverlines between peristome and aboral trochal band 20-24, from aboral trochal band to scopula 9-11; abstomal end of row 1 of infundibular polykinety 3 diverges from the other two rows of this polykinety and ends alongside row 3 of infundibular polykinety 2. PMID- 17113474 TI - The microfauna communities and operational monitoring of an activated sludge plant in China. AB - The composition of the microfauna community in aeration tanks at the Baoding Sewage Treatment Plant in China was analysed each week from July 2002 to July 2003. The community composition of these microfauna populations was compared with effluent quality data recorded on the same days. A total of 94 species of ciliates, 40 species of amoebae and 13 species of large flagellates were identified in the 50 samples analysed. Numbers of metazoa including rotifers, nematodes, gastrotrichs and oligochaeta were also recorded. Although, Aspidisca cicada showed the highest mean abundance and was present in 98% of the samples, factor analysis revealed, among other things, that high Vorticella convallaria and Arcella hemisphaerica populations correlated with good performance of the treatment plant, while high numbers of Litonotus obtusus indicated poor conditions for settlement of sludge. These results show some agreement with an earlier study of sewage plants in Beijing, but analysis of more plants having a diversity of input components working under a range of different operating conditions should be performed to gain a general understanding of the value of indicator species for predicting the efficiency of activated sludge plants in China. PMID- 17113475 TI - New entodiniomorphid ciliates from the intestine of the wild African white rhinoceros belong to a new family, the Gilchristidae. AB - Gilchristia artemis n.g., n.sp. and Digilchristia draconis n.g., n.sp. in the order Entodiniomorphida are described from the large intestine of the African white rhinoceros, and a new family Gilchristidae is proposed to contain them. These new species have a C-shaped adoral polybrachykinety, a slender vestibular polybrachykinety, and paralabial kineties along the ventral side of the adoral polybrachykinety in their retractable adoral ciliary zone, showing the same arrangement as in the rumen ciliates in the family Ophryoscolecidae. G. artemis has two skeletal plates and D. draconis one plate. In both species the dorsal skeletal plate is bow-shaped, folded in half longitudinally, twisting in the anterior part, and lying along the dorsal left side of the macronucleus. The second plate of G. artemis is slender and lies along the ventral side of the macronucleus. G. artemis has three ciliary arches and D. draconis has four arches along the dorsal and ventral sides of the body. Their arches are long and non retractable, closely resembling those of ciliates in the families, Spirodiniidae and Cycloposthiidae, and are not analogous to the single retractable ciliary arch of the rumen ciliates in the family Ophryoscolecidae. PMID- 17113476 TI - Newly discovered linkages between the cortical (pellicular) ridges of Opalina. AB - Samples of Opalina ranarum have been prepared for electron microscopy by ultra rapid cryofixation followed by substitution fixation in a solvent containing tannic acid. This technique has made it possible to see that very thin linkages exist between the pleated ridges that form the surface of the cell. Between any two adjacent cortical ridges, the linkages, which are approximately 0.1 microm long, occur as a single row, 0.1 microm below the free edge, with an impressively regular spacing of 0.1 microm. The cortical ridges of the Opalinids are spaced with remarkable uniformity, even when thrown into undulating patterns. The linkages described here will inevitably stabilize the complex architecture of the cortex. Other possible functions are discussed. PMID- 17113477 TI - Ultrastructural study of developmental stages of Mattesia dispora (Neogregarinorida: Lipotrophidae), a parasite of the flour moth Ephestia kuehniella (Lepidoptera). AB - The ultrastructure of merozoites, gamonts and oocysts of the neogregarine Mattesia dispora and their development in larvae of the flour moth Ephestia kuehniella were studied by electron microscopy. The apical complex of free macronuclear merozoites was very distinct in micrographs of sections, the polar rings being especially prominent. Two gamonts associated in head-to-head syzygy and the apical complexes served as the contact point during pairing. At this stage the rhoptries became reduced and the conoid widened. The gamonts had a foam like appearance in the light microscope. Paired gamonts formed an envelope and developed into a gametocyst, within which the gamonts were separated by a distinct border. Four gametes and two residual cells developed inside the gametocyst. The gametes were covered with a single membrane. The gametes fused in pairs to form two spherical zygotes, each covered by two membranes and with one large nucleus. The external layer appeared more undulated than the inner one. A single membrane covered each residual cell. Walls were formed around both zygotes to produce two oocysts. Each mature oocyst was lemon-shaped with polar plugs and eight peripheral sporozoites, which had a pellicle similar to that of the merozoites, lay beneath the thick oocyst wall. PMID- 17113478 TI - The impact of HIV/AIDS on men's health: an action imperative. PMID- 17113480 TI - Prevention with HIV-infected men: recommendations for practice and research. AB - In the United States in 2004, 74% of the new AIDS cases and 70% of the new HIV cases were in men; in addition, 75% of the cases of HIV in women were classified as heterosexually acquired. These numbers make it clear that expanded prevention efforts for men who are infected with HIV would make a large contribution to containing the epidemic. This report explores epidemiologic and psychosocial issues related to prevention in men with HIV and compares how those variables relate to prevention efforts. The report ends with a discussion of a method to approach HIV risk reduction in clinical care settings. PMID- 17113479 TI - A portrait of HIV infection among men in the United States. AB - HIV/AIDS has been a major public health problem in the United States for over 25 years and has significantly contributed to morbidity and mortality among men. At the onset of the HIV/AIDS epidemic, White men were predominantly affected, representing 95% of the cases in 1982. Over time, the burden of disease shifted from White men to Black and Hispanic men. Currently, Blacks and Hispanics represent 64% of the men living with HIV. AIDS is the sixth leading cause of death for Black men and the tenth leading cause of death for Hispanic men. Because the transmission of HIV is predominantly behavior-driven, it is imperative that nurses have open, honest discussions with clients about their behaviors. PMID- 17113481 TI - Understanding the HIV risk reduction needs of heterosexual African American substance-abusing men. AB - With the increasing prevalence of HIV/AIDS, nurses find themselves caring for diverse populations at risk for HIV. One subpopulation at risk is African American men with a history of substance use. To better understand the risk reduction needs of these men, a focus group was conducted with 16 African American men attending an outpatient drug treatment program in Philadelphia. The purpose was to identify perceptions of HIV risk, engagement in HIV risk behaviors, and barriers to condom use in order to generate recommendations for risk reduction programs tailored to the needs of this population. Results revealed that African American substance-abusing men perceive themselves to be at risk for HIV infection and other adverse health outcomes yet lacked adequate information related to HIV prevention. The need for culture- and gender-specific interventions to reduce HIV-related risk behaviors among African American substance-using men is discussed. PMID- 17113482 TI - Disclosure decisions of rural African American men living with HIV disease. AB - A qualitative study was conducted to explore disclosure decisions of rural African American men living with HIV disease. The sample consisted of 20 HIV infected African American men living in the rural South who had been diagnosed with HIV for at least 6 months. Audiotaped semistructured interviews were used for data determination. The men were questioned about who they had told about their disease, reactions to their disclosures, and their advice to others about disclosing. Results showed that initially the men did not disclose their disease to others, and many of them continued not to disclose. They were concerned about negative consequences such as rejection, fear of contagion, and the recipients telling others. If and when they disclosed, it was likely to be to sexual partners, immediate family members, and health care providers. Their decision not to disclose protected them from the possible negative reactions, but it also limited the amount of social and emotional support they received related to their HIV disease. PMID- 17113483 TI - Conducting nursing research with men who have sex with men: challenges and strategies for nurse researchers. AB - The group most affected by the HIV epidemic is men who have sex with men (MSM). Since the beginning of the epidemic in 1981, this group was one the four identified high-risk groups, and MSM continue to comprise nearly 50% of all cases of HIV/AIDS. In the context of HIV infection and safer sex behaviors, this population has been the focus of numerous research studies. Despite the wealth of research that has been conducted on this population, very little information is available on research methods and strategies that nurse researchers can use to study this population. This report details one nurse researcher's experiences in gaining access to this population, unique recruitment issues, and challenges in data collection. In addition, strategies and interventions that were used by this researcher to overcome these challenges in the research process are discussed. PMID- 17113484 TI - Internet chat rooms: connecting with a new generation of young men of color at risk for HIV infection who have sex with other men. AB - The purpose of this study was to explore the use of gay-related Internet chat rooms by young men who have sex with other men of color (YMSMC) in a specific catchment area. Participants were 104 YMSMC age 18 to 24 (M = 21.56) who were encountered in two gay-related Internet chat rooms during April 2005. Participants were mainly African American (53.7%, n = 56), HIV-negative (57.6%, n = 60), and online looking for some type of sexual encounter (80.7%, n = 84). The results of this study support the need to develop specific culturally appropriate HIV prevention Internet outreach protocols targeting YMSMC at risk for HIV infection. Lessons learned while conducting this study and recommendations are also discussed. PMID- 17113485 TI - Men's health: it is not just about HIV anymore. PMID- 17113486 TI - Where were the doctors? Torture and the betrayal of medicine. PMID- 17113488 TI - Trends that will affect your future ... where were you? PMID- 17113489 TI - Glyconutrients: the state of the science and the impact of glycomics. AB - The science of glycobiology has made rapid strides in the past few decades. A recent development has been the introduction of glyconutrients as nutritional supplements for health support and disease management. This article reviews the basic and clinical science of glyconutrients and presents a brief perspective on the impact of the field of glycomics. PMID- 17113490 TI - Positive emotional change: mediating effects of forgiveness and spirituality. AB - We evaluated the efficacy of an emotional education program that seeks to reduce the intergenerational transmission of negative interaction patterns by increasing forgiveness and spirituality. We examined both reduction of psychological symptoms and increase in positive psychological outcomes over the course of a year, as well as the mediators of this change. At baseline, the sample consisted of 99 participants and 47 waiting list controls. Comparisons of scores from baseline (Time 1) to one week after the Hoffman Quadrinity Process (Time 2) showed large declines in negative affect (depressive symptoms) and increases in both positive outcomes (mastery, empathy, emotional intelligence, life satisfaction, forgiveness, and spiritual experience) and health and well-being. Over the course of a year, most of these gains were sustained, in comparison with the control group. Further, increases in forgiveness and spirituality mediated the effect of program participation on depressive symptoms. PMID- 17113491 TI - World hypotheses and the evolution of integrative medicine: combining categorical diagnoses and cause-effect interventions with whole systems research and nonvisualizable (seemingly "impossible") healing. AB - It has been proposed that to understand (1) the evolution of science and medicine, and (2) the integration of conventional, complementary and alternative medicine, it is essential to consider at least eight universal implicit meta cognitive hypotheses. It has been suggested that these implicit "world" hypotheses can be applied in every discipline of science. The present paper reviews the eight world hypotheses and proposes an additional hypothesis, termed the nonvisualizable or "Nth" world hypothesis (adopting the mathematical concept of "N"; eg, as in N dimensional space). Drawing on contemporary mathematics and quantum physics, we propose that certain theories and data-by their inherent nature-can not be visualized, and therefore may seem "unimaginable" and "impossible" (if not "unbelievable"), even though they are real. Certain seemingly anomalous observations in mind-body and energy medicine, including areas historically labeled as parapsychology or spiritual energy healing, often elicit strongly skeptical and dismissive reactions. We propose that these skeptical and dismissive reactions to purportedly impossible (yet logical) theories and seemingly unbelievable (yet replicable) data can be tempered when the Nth world hypothesis is understood and incorporated. Integrity in evidence based science and medicine may require that scientists and nonscientists alike develop comfort and humility in accepting the human mind's restricted ability to envision and imagine certain nonvisualizable-yet fundamental and real-concepts and effects, as illustrated in contemporary physics and complementary and alternative medicine. PMID- 17113492 TI - Disease of distinction. AB - Gout is one of the rare diseases that defines its sufferers by class and culture. It is also one of the first chronic diseases to be clinically described. The Egyptians had identified gout as a distinct disorder by 2640 bce. This paper traces the history of gout from its earliest recorded period down to modern times, with a particular emphasis on the cultural, political, geopolitical, and social aspects. Included is a discussion of its role in the American Revolution. Today, gout is a well-understood clinically managed arthritic disease that excites little comment. This is an entirely modern perspective. For most of human history, gout was a disease of distinction that dominated much of medicine, playing the same role in Rome's third century bce aristocracy it would later play in the aristocracies of 17th and 18th century France and England, when each of these countries dominated the world. Because it was considered a disease of lifestyle until modern times, when genetics began to be understood, gout was associated with rich, high status Caucasian men and their excessive consumption of drink and rich foods. It was virtually unknown in Asia, until Western dietary practices became widespread there. From earliest history, gout has been linked with high IQ and sexual promiscuity, which made it grist for artists and writers, and their social commentary up to the time of Dickens; this is discussed, with examples. Because of its association with the rich, gout also developed a powerful moralistic aspect, particularly during the Christian era when the concept of sin was a cultural fundamental. The loose living and indulgence of the rich and the gout it produced made the disease a parable of Christian ethics. The Italian poet Francesco Petrarch (1304-1374) was one of the first to establish this nuance, and it influenced how gout was seen for centuries. Part of what gave gout its special character was that while it tortured, it rarely killed. Indeed, when death was a frequent visitor to families, it was thought a painful but welcomed prophylactic against diseases that did kill. Even in modern times, gout still favors the rich and powerful. American research conducted in the 1960s found that corporate executives, just like their English gentry, or Roman senatorial predecessors, had higher urate concentrations than their blue-collar employees. PMID- 17113494 TI - The integrative family practice. An interview with Ben Kligler by Bonnie J. Horrigan. PMID- 17113493 TI - Fever and high C-reactive protein level as the sole manifestation of allergy to cow milk. PMID- 17113495 TI - Potential health benefits of green tea (Camellia sinensis): a narrative review. PMID- 17113496 TI - Spiritual assessment: a chaplain's perspective. AB - Every hospital and medical center is a dynamic, shifting terrain; each develops its own culture. Even within a single hospital system that is attempting to integrate a strong mission statement, providers routinely experience differences in culture between departments, floors and campuses! These differences are characterized largely by the quality of the past history of the organization combined with the human beings currently working together as staff. Even though the cultural sands are continually shifting, Chaplain Larry Austin's voice is clear, unburdened and practical. He identifies a problem of lack of collaboration between health care professionals of diverse disciplines, as well as a resultant lack of understanding and specificity of professional functions and concepts. These include the challenge of shared languages. While each of the powerful realities identified below offers readers the opportunity for further reflection and dialogue, his starting points for standards in spirituality in health care are symphonic; they stress the wisdom of collaboration between disciplines. Music to the integrative medicine ear! PMID- 17113497 TI - Integrating tuina acupressure and traditional Chinese medicine concepts into a holistic nursing practice. PMID- 17113498 TI - Innovations in integrative healthcare education: mind-body faculty development at UCLA and the symposium for portland area research on complementary and alternative medicine. AB - Content on integrative healthcare and complementary and alternative medicine (CAM) is being taught in hundreds of educational programs across the country. Nursing, medical, osteopathic, chiropractic, acupuncture, naturopathic, and other programs are finding creative and innovative ways to include these approaches in new models of education and practice. This column spotlights such innovations in integrative healthcare and CAM education and presents readers with specific educational interventions they can adapt into new or ongoing educational efforts at their institution or programs. We invite readers to submit brief descriptions of efforts in their institutions that reflect the creativity, diversity, and interdisciplinary nature of the field. Please submit to Dr. Sierpina at or Dr. Kreitzer at . Submissions should be no more than 700-800 words. Please include any Web site or other resource that is relevant, as well as contact information. PMID- 17113500 TI - The management of life-threatening haemorrhage following blunt facial trauma. AB - Life-threatening bleeding is uncommon following blunt facial trauma. There have been few reports in the literature describing its optimal management and a clear approach to treatment is yet to be defined. Reported strategies for control of facial haemorrhage include oro-nasal packing, external carotid artery ligation, transantral ligation of the internal maxillary artery, maxillary reduction and angiographic embolisation. Advances in angiography and selective vessel embolisation have made this the treatment of choice in cases of bleeding following penetrating facial injury. Its use in the management of bleeding following blunt facial trauma is unclear. The combined experience of the facial trauma teams at Harborview Hospital, Seattle, USA and Liverpool Hospital, Sydney, Australia yielded four cases of severe life-threatening haemorrhage following blunt trauma that underwent angiography. The roles of various management strategies were evaluated to generate a preferred management pathway in treating severe bleeding following blunt facial trauma. PMID- 17113501 TI - The effect of surgical resection in the region of the retromolar trigone. AB - Tumours in the mucosa of the retromolar trigone (RMT) are rare, but develop insidiously and spread rapidly into surrounding structures. Resection may require radical dissection beginning usually on the medial side of the mandible. Such surgery can put important structures at risk. The normal anatomy of the RMT and its relations has been studied together with simulated surgical incisions and resections. Tissue removed was processed by histological techniques in order to demonstrate structures excised or damaged by the operation. The simulated incision showed that the lingual nerve, submandibular duct and palatoglossus were at particular risk. This could affect sensation, speech, swallowing and movements of the tongue. The findings pose immediate concerns for surgeons operating in this area. Although cancer surgery often involves sacrificing tissue, care should be taken to preserve structures vital to the patient's oral function without compromising oncological principles. PMID- 17113502 TI - Voice reconstruction using the free ileocolon flap versus the pneumatic artificial larynx: a comparison of patients' preference and experience following laryngectomy. AB - This study compares the psychological adjustment and voice function of patients undergoing voice rehabilitation using the free ileocolon flap for creation of a voice tube shunt and patients undergoing voice rehabilitation using the pneumatic artificial larynx. Twelve laryngectomy patients were included; six underwent free ileocolon transfer following a period of pneumatic artificial larynx use. Mean duration after laryngectomy was 5.2 years. Mean follow-up was 210 days. A chart review, questionnaires and a prospective evaluation were performed. Voice tube shunt patients had better speech function and higher self-esteem. People's discrimination and appearance when speaking were important in the patients' choice of method for rehabilitation. There was a high preference for choosing the voice tube shunt and a higher motivation and willingness to use that voice mechanism in the voice tube shunt group. Patients who undergo free flap reconstruction of voice have better speech function and self-esteem than patients who continue to use the external pneumatic device. Psychological assessments are important for surgical patients in order to evaluate a critical aspect of our perceived success - the patients' perception. PMID- 17113503 TI - Experience with the modified hockey stick incision for block dissection of neck. AB - BACKGROUND: The modified hockey stick incision was originally described by Lahey in 1940. This allows elevation of a superiorly based cervical skin flap, with additional exposure of the parotid as necessary. METHODS: The longitudinal portion of the incision runs from the mastoid process downward, behind the anterior border of the trapezius muscle, and curves gently at the junction of the lateral one-third and medial two-thirds of the clavicle. The transverse component extends medially, approximately 2 cm below the clavicle. When parotidectomy is required, incision is extended in front of the ear and a forward cut is made for additional exposure. RESULTS: We present our experience with a series of 16 patients, undergoing a total of 17 neck dissections for various pathologies. The majority (11 patients) received postoperative radiotherapy. There were two episodes of minor skin necrosis which were not related to radiotherapy, but to patient co-morbidity, and settled with conservative management. All patients had a satisfactory cosmetic result. CONCLUSION: We find the modified hockey stick incision to be cosmetically superior and provide excellent exposure of the neck, with protection of the carotid vessels. PMID- 17113504 TI - Periocular metatypical cell carcinoma: clinicopathologic correlation, management, and follow-up in 35 patients. AB - BACKGROUND: To present the clinical characteristics and treatment outcomes in a relatively large series of patients with periocular metatypical cell carcinoma. METHODS: This is a retrospective study of 35 patients with periocular metatypical cell carcinoma, who were seen between January 2000 and December 2004 in Izmir Ataturk Research and Training Hospital, Turkey. The patients were histologically confirmed metatypical cell carcinoma with radiologic evidence of periocular region. The main outcome measures were patients' demographics, duration of tumour, clinical presentation, tumour site, treatment modalities, recurrence rate and tumour-related exenteration. Statistical comparisons between the nonrecurring group and recurring group were completed using the Cox regression analysis and the log rank test. Significance was determined for all statistical tests as pe 0.05. RESULTS: The median age of patients at time of diagnosis was 61 years. The most common site for metatypical cell carcinoma was medial canthus and lower eyelid region. The median follow-up for all patients was 18 months. The average length of time until recurrence was 13 months. Five of the 35 patients received adjuvant therapy to the primary tumour site after excision. Signs suggestive of orbital involvement included bone fixation of the mass, limitation of ocular motility and globe displacement. The early lack of diagnostic criteria for metatypical cell carcinoma created considerable debate and confusion regarding its diagnosis. We believe that once this diagnosis is made, the treatment modality should be individualized and chosen with consideration of the extent of orbital involvement, visual function, and the patient's general health. PMID- 17113505 TI - Parotid metastasis--an independent prognostic factor for head and neck cutaneous squamous cell carcinoma. AB - BACKGROUND: Metastatic parotid cutaneous squamous cell carcinoma (SCC) is the most common parotid gland malignancy in New Zealand and Australia. The current AJCC TNM staging system does not account for the extent of nodal metastasis. A staging system that separates parotid (P stage) from neck disease (N stage) has been proposed recently. AIM: To review the outcome of patients with metastatic head and neck cutaneous SCC treated at our multidisciplinary Head and Neck Service using the proposed staging system. METHOD: Consecutive patients were culled from our Head and Neck/Skull Base Database, 1990-2004. These patients were restaged according to the proposed staging system: P stage: P0 = no disease in the parotid (i.e., neck disease only); P1 = metastatic node < or = 3 cm; P2=metastatic node > 3 cm and < or =6 cm, or multiple nodes; and P3 = metastatic node > 6 cm, or disease involving the facial nerve or skull base. N stage: N0=no disease in the neck (i.e., parotid disease only); N1 = single ipsilateral metastatic node < or = 3 cm; and N2 = multiple metastatic nodes, or any node > 3 cm, or contralateral neck involvement. Loco-regional recurrence and disease specific survival were calculated using the Kaplan-Meier method and comparison of graphs made with the log-rank test. Multivariate analysis using the Cox regression model was carried out to assess the impact of various parameters. RESULTS: Sixty-seven patients with metastatic head and neck cutaneous SCC were identified. Thirty-seven patients had parotid metastasis (of whom 13 also had neck disease) while 21 had neck metastasis alone. Nine patients had dermal or soft tissue metastasis. These nine patients were excluded from this series, and data analysis was carried out on the remaining 58 (46 men, 12 women, mean age 71 years) patients. Sixty-seven percent of the patients underwent post-operative adjuvant radiotherapy. The five-year disease-specific survival rate was 54%. Among 56 patients followed up to disease recurrence or for a minimum period of 18 months, the loco-regional recurrence rate was 52%. The presence of parotid disease was an independent prognostic factor on survival (p < 0.01), and P3 fared significantly worse than P1 and P2. Those patients who had both parotid and neck disease fared worse than those who had parotid or neck disease alone (p = 0.01). N2 had a significantly poorer outcome compared with N1 (p < 0.01). Immunosuppression (p = 0.01) and a positive surgical margin (p < 0.01) were significant adverse prognostic factors for survival. Adjuvant radiotherapy, extracapsular spread, and perineural and vascular invasion did not influence survival. Our study demonstrates that the extent of parotid disease is an independent prognostic factor for metastatic head and neck cutaneous SCC. PMID- 17113506 TI - Merkel cell carcinoma: a report of 34 cases and literature review. AB - BACKGROUND: Merkel cell carcinoma (MCC) is a rare and aggressive skin cancer, with unclear histogenesis. To date there is no consensus on the optimal treatment of this neoplasm, with controversy surrounding the use of radiotherapy and chemotherapy. There are also limited data on biological behaviour and prognosis, with reported survival ranging from 31% at three years to 74% at five years. METHOD: The medical records of 34 patients with a diagnosis of primary MCC, treated at two NHS trusts in Birmingham and Coventry, were reviewed. An extensive review of the English literature was also performed. RESULTS: MCC occurred predominantly in Caucasians (97%) with a mean age of 75 years. Identified risk factors were a previous history of SCC (37%), BCC (18%) and AK (20%). Ten percent of patients showed evidence of immunocompromise. Most tumours were located on the extremity, where they reached a mean size of 2.1cm. Fifty percent had regional metastasis during the course of their disease. A sub group analysis of the excision margins showed that a 2-cm excision margin, extending to the deep fascia, resulted in a 50% incomplete excision rate and a 33% local recurrence rate. In contrast a 3-cm margin including deep fascia resulted in no incomplete excisions and a 10.5% local recurrence rate Prognosis was poor with a 40% 3-year survival. Combining the data from two trusts has produced a relatively large series and highlighted differences in patient characteristics and management between the units. We advocate a 3-cm excision margin, including fascia wherever possible, combined with post-operative radiotherapy to offer the best chance of local control. Survival is fairly dismal and in keeping with the aggressive nature of this tumour. The respective roles of radiotherapy and chemotherapy remain controversial. PMID- 17113507 TI - Microvascular reconstruction of nasal ala using a reversed superficial temporal artery auricular flap. AB - Microsurgical technique allows successful transfer of an auricular flap in a one stage procedure, using the root of the helix. Although a free composite auricular flap with the superficial temporal artery pedicle provides a good solution to repair nasal defects, its vascular pedicle is so limited that a vein graft from other area of the body is usually needed to reach the recipient site, leaving an unpleasant scar on the donor site. The authors present a reversed superficial temporal artery auricular free flap for alar reconstruction by microsurgical transfer. This technique has been performed on four patients with posttraumatic alar defects. In three patients, the reversed superficial temporal vessels of the flap were anastomosed directly with the recipient facial vessels in the nasolabial fold. In one patient, the reversed superficial temporal artery of the flap was anastomosed with the facial artery as above, its accompanying vein to the proximal stem of the superficial temporal vein by a graft taken from the excess length of the reversed superficial temporal artery pedicle because a suitable vein was not found for microvascular anastomosis in the nasolabial area. In these four patients, the size of the flap was 2.5 x 2.0-4.0 x 2.5 cm, the length of the vascular pedicle is 5-8 cm, average 6.5 cm. The reversed superficial temporal artery auricular flap offers a long vascular pedicle of the auricular free flap for microvascular anastomosis in the reconstruction of the ala of nose, delivers a good solution to the problem of the vascular pedicle shortage of the proximal superficial artery auricular flap. There is no need of vein graft from other parts of the body because the superficial temporal vessels on the temple provide not only the flap pedicle but also a source of vessel grafts. This technique may have even wider applications in other facial cutaneous defect. PMID- 17113508 TI - Reconstruction of orbital floor and maxilla with divided vascularised calvarial bone flap in one session. AB - We present four cases which underwent reconstruction of orbital floor and anterior maxillary wall with a vascularised bone flap following partial maxillectomy. After tumour resections, superficial temporal artery (STA) and vein based calvarial bone flaps from the outer tabula were prepared. Without disrupting the integrity of fascia and periosteum, the bone was separated into two segments in the same direction as the blood flow and one is 3 cm and the other 5 cm. The two bone segments were transferred as one single flap and one segment of the flap was used to reconstruct the orbital floor and the other for reconstruction of the anterior maxillary wall. Since two cases had large skin defects, lateral frontal skin to which the frontal branch of the STA supplies blood was incorporated into the flaps. Functional and aesthetic results were satisfactory at the end of 8-20 months follow-up. This technique allowed reconstruction of the orbital floor and anterior maxillary wall and even skin defects with a single pedicled flap in one session. PMID- 17113509 TI - Periodic alterations of jejunal mucosa morphology following free microvascular transfer for pharyngoesophageal reconstruction. AB - BACKGROUND: Free jejunal flap reconstruction is the treatment of choice for patients after pharyngoesophagectomy. It remains unclear as to how the transplanted jejunal mucosal damage proceeds after the warm ischaemia. The current study aims to assess the relationship between the duration of ischaemia and the damage of jejunal mucosa. PATIENTS AND METHODS: From May 2002 to February 2003, 15 free jejunal flaps in 15 patients were transplanted to the cervical area for the reconstruction after pharyngoesophagectomy. Biopsy specimens were taken from the monitor loop at the time of pedicle ligation, 10 min after reperfusion, every day for 10 days, 14th day, 28th day, and 40th day after operation. Mucosal injury was assessed based on an accepted three-point scale which evaluates oedema, inflammation, mucosal necrosis or exfoliation, shortening of villi, and increase of goblet cells. FINDINGS: All 15 jejunal flaps survived. The mean ischaemia time was 68.7+/-5.2 min (range: 37-116). Serious injury to the mucosa was observed at 10 min after reperfusion, and gradually recovered until the 8th day, when it became normal in all flaps. The degree of damage was not found to be correlated with the length of ischaemia (less than 116 min). Severe ischaemia/reperfusion-induced mucosal damage occurs immediately following reperfusion and gradually recovers with time. The severity of the damage is not related linearly to the ischaemia time within 2h. The mucosa recovers gradually from the 8th day and returns to normal at the 28th day. PMID- 17113510 TI - A simple classification for standardisation of nomenclature in free flap outcome. AB - The numbers of free flap donor site as well as their indications are constantly increasing. Despite increasing popularity of microvascular reconstructive procedures, literature lacks clear and objective outcome criteria. This paper reports on a simple outcome classification that has become a routine part of the unit's large workload of microvascular outcome recording. The classification was formed through a retrospective analysis of 241 consecutive cases from 2000 to 2001 and is a five graded numerical classification. Grade 1 equates to total success without co-morbidity and grade 5 to a major complication such as amputation, etc., whatever the status of the flap itself. From 2002 to 2005 the classification was prospectively used on 527 consecutive cases with ease of integration into routine clinical practice. The Classification would enable a more objective record keeping thus analysis of the outcome. It would allow a more realistic comparison of different techniques or donor types as well set a benchmarking level for further improvement of the results. PMID- 17113511 TI - Giant combined microdissected thin thigh perforator flap. AB - Despite recent advances in reconstructive surgery, extremely wide and thin flap coverage has rarely been reported. Recently, the authors developed a technique for transferring a very wide and thin flap from the thigh area using microdissection. In this procedure, both perforators of the anterolateral thigh flap and the tensor fasciae latae perforator flap were microdissected simultaneously and these two perforator flaps were elevated in combination based on the common pedicle of the vessels. In this report, the detailed technique of the procedure is described along with a discussion of the safety of this giant flap in consideration of 60 clinical experiences of microdissected thin tensor fasciae latae perforator and anterolateral thigh flaps. PMID- 17113512 TI - Further application of the bilobed flap: the split bilobed flap for reconstruction of composite posterior auricular and mastoid defects. AB - In this article a modified bilobed flap from mastoid and lateral neck skin for reconstruction of complex defects of the posteromedial surface of the auricle and mastoid skin, with the preservation of the retroauricular sulcus, is described. Reconstruction of the postero-medial auricular surface has almost never been a concern for reconstructive surgeons. It is in fact a shaded area with little aesthetic relevance and direct closure, skin grafting and even secondary healing are used for skin cancer defects repair. Also mastoid skin defects can be repaired with simple techniques such as skin grafts or transposition flaps from the remaining mastoid skin or from the neck. On the other hand, cancers involving the postero-medial auricular surface, the retroauricular sulcus and the mastoid skin require wide and deep resections that involve the posterior auricular muscles and reach the perichondral and periosteal surfaces. Direct closure with undermining, if feasible, will obliterate the retroauricular sulcus causing asymmetry with the contralateral ear and, if defects are cephalad will impair the possibility of wearing spectacles, thus leaving functional and aesthetic impairment. Transposition flaps from the remaining mastoid skin, due to the lack of skin laxity, are not feasible because the donor site cannot be closed. Two patients, both affected by basal cell carcinoma involving the posteromedial auricular surface and the mastoid skin have been treated with this flap. In both cases the use of the modified bilobed flap described in this article allowed preservation of the retroauricular sulcus and closure of the donor site. Scars were hidden along minimal tension lines and the possibility of wearing spectacles along with sensitivity all over the reconstructed area were maintained. PMID- 17113513 TI - Reversed anterior interosseous flap. AB - Reversed flaps from the forearm have been firmly established for hand coverage. Each has its own advantages and disadvantages. The reversed anterior interosseous flap is one option with special advantages in hand reconstruction. From January 2002 to July 2003 we used this flap in five consecutive male patients aged between 36 and 59. The defects were located on the first web space (2), on the dorsal side of the thumb (1), on the first metacarpal bone (1), and on the volar side of the wrist (1). Flap size was between 6 x 3 cm and 11 x 7 cm. All flaps healed without any problem and no complication was observed during the postoperative period. The major advantage of this flap is the preservation of the main arteries of the upper limb. Other advantages are as follows: good texture and colour for hand reconstruction, satisfactory rotation arc, and availability of composite and fascial flaps. Besides its advantages, the major disadvantages are unsightly donor area scar and the need for meticulous technique. As a conclusion, we found this flap very useful in hand reconstruction for coverage of small and moderate sized soft tissue defects of the hand. PMID- 17113514 TI - A new test for demonstrating the action of flexor digitorum superficialis (FDS) tendon. AB - The available tests for demonstrating the action of flexor digitorum superficialis (FDS) can be used on one finger at a time. A new test is proposed which can test all the four FDS tendons simultaneously. It was administered along with other available tests on 100 uninjured hands of 50 volunteers and on seven clinical cases with FDS injury. It correlated fully (100%) with the standard test on middle and ring fingers and with the pulp-to-pulp forced pinch test with the thumb on index fingers. It was more accurate than the standard test in detecting the presence of FDS action on the little finger. It was found to be in strong agreement with the modified standard test on little finger with a kappa coefficient of 0.98. It correlated fully for the absence of FDS tendon action in seven operatively proved cases. The present test is a new test which is faster and more versatile than the available tests. PMID- 17113515 TI - Architecture of European plastic surgery. AB - The architecture of European Plastic Surgery was published in 1996 [Nicolai JPA, Scuderi N. Plastic surgical Europe in an organogram. Eur J Plast Surg 1996; 19: 253-256.] It is the objective of this paper to update information of that article. Continuing medical education (CME), science, training, examination, quality assurance and relations with the European Commission and Parliament all are aspects covered by the organisations to be discussed. PMID- 17113516 TI - Internal thoracic vessels as recipient vessels for free flap reconstruction in head and neck surgery. AB - In rare cases the usage of the internal thoracic vessels as recipient vessels in reconstructive surgery of the head and neck region with free tissue transfer is a challenging but valid alternative if local recipient vessels are unusable. PMID- 17113517 TI - Maxillary reconstruction using a free deep inferior epigastric perforator (DIEP) flap combined with vascularised costal cartilages. AB - In this article, we describe a case of maxillary carcinoma requiring reconstruction in which we used a free deep inferior epigastric perforator (DIEP) flap combined with vascularised costal cartilages. The DIEP flap was harvested with a rectus abdominis muscle in its cranial side. Eighth and ninth costal cartilages were harvested and connected with the muscle. The Zygomaticomaxillary buttress was reconstructed using vascularised costal cartilages. Nasal and oral lining were reconstructed with a DIEP flap. The viability of grafted bone was proved in bone scintigraphy and three-dimensional computed tomography after six months. Cosmetically the result was good. Compared with other methods, this flap can be thinned easily to match a defect and vascularised cartilages connected with rectus muscle can be nourished by the same vascular pedicle. This is a first report of perforator flap combined with vascularised cartilage. This flap has a possibility to be used for a lot of reconstruction that needs bone reconstruction with perforator flap. PMID- 17113518 TI - Eleven-year survival from an intra-dermal melanoma. AB - Stage IV metastatic malignant melanoma of unknown primary (TxNxM1a) is known to have a poor prognosis. However, some patients suffering from cutaneous disease originally thought to represent metastasis have fared much better than expected. We report a patient who has survived 11 years following such a diagnosis. Due to the prolonged survival and absence of an identified primary, it is unlikely that the lesion was metastatic but may represent one of a number of other possibilities. A small number of similar cases in the literature suggest a need for awareness of this unusual group of patients. PMID- 17113519 TI - Multiple malignant melanomas in association with neurofibromatosis type 1. AB - We present a case of multiple primary malignant melanomata occurring over a six year period in a 63-year-old Caucasian man with neurofibromatosis type 1. There is doubt regarding a definite association between these two diseases despite a number of case reports and clear, potential pathological mechanisms. This case not only strengthens support for an association but also highlights the great difficulties that arise in the management of cutaneous melanomata in patients with neurofibromatosis. PMID- 17113520 TI - Successful management of recalcitrant groin lymphorrhoea with the combination of intraoperative lymphatic mapping and muscle flap. AB - Recalcitrant groin lymphorrhoea in high-risk patients remains a problem. In this report, a cardiac transplant patient with recalcitrant groin lymphorrhoea was successfully treated with a combination of intraoperative lymphatic mapping and sartorius muscle flap. We believe that the combined use of these two treatment options offers a more effective approach for surgical treatment of recalcitrant groin lymphorrhoea and should be considered when managing this difficult clinical problem, especially in high-risk patients. PMID- 17113521 TI - Post-traumatic and post-surgical Absidia corymbifera infection in a young, healthy man. AB - Absidia corymbifera infection in a healthy individual is rare. Most of the infection occurs in immunocompromised patients or diabetic patients. Cutaneous and subcutaneous mucormycosis have been increasingly reported in the literature as a result of massive trauma with contaminated wounds. We present a case of cutaneous mucormycosis in a healthy, young patient after surgical amputation for a crush injury of the leg. We also highlight the importance of the high index of clinical suspicion in the diagnosis and treatment of this fungal infection in the hype of methicillin-resistant Staphylococcus aureus (MRSA) infection in hospital setting these days. Despite an initial life-saving amputation, it was inadequate to ensure the eradication of A. corymbifera infection. A second amputation was required with parenteral liposomal amphotericin B to achieve a satisfactory cure. PMID- 17113522 TI - Extensive actinomycosis of the face requiring radical resection and facial nerve reconstruction. AB - We present a case of extensive actinomycosis of the face, which appeared after dental surgery. Since antibiotic therapy was ineffective, the lesion was radically resected, and the skin, soft tissue and facial nerve were reconstructed using a free rectus abdominis musculocutaneous flap and simultaneously harvested intercostal nerves. Successful reanimation of the face was achieved 14 months postoperatively. PMID- 17113523 TI - Pneumococcal septicaemia with Purpura fulminans in an 11-month-old child. AB - Purpura fulminans (PF) is a syndrome characterised by acute onset of rapidly progressive haemorrhagic necrosis of the skin due to dermal vascular thrombosis, mainly occurring during meningococcal sepsis. It occurs rarely in the course of infection with Streptococcus pneumoniae and most cases report Meningococcus as the causing agent. This is a case report of successful conservative limb preserving management of PF and sepsis caused by Streptococcus pneumoniae in an 11-month-old girl. PMID- 17113524 TI - Bacteraemic necrotizing fasciitis with compartment syndrome caused by non-O1 Vibrio cholerae. AB - Non-O1 Vibrio cholerae are gram-negative rods that can cause sporadic gastroenteritis, bacteraemia, and extraintestinal infections, primarily following the consumption of raw seafood or exposure of damaged skin to contaminated saltwater during the summer months. Bacteraemic necrotizing fasciitis caused by non-O1 V. cholerae has rarely been reported. Liver cirrhosis, haemochromatosis, and immunosuppression are important factors contributing to the severity of the infections and outcome. This report describes a case of liver cirrhosis in which right lower leg compartment syndrome and acute renal failure presented as the initial symptoms of bacteraemic necrotizing fasciitis. The organisms growing in the wound and blood cultures were identified as non-O1 V. cholerae. After antibiotic therapy, fasciotomy, right above-knee amputation, repeat debridement of the left lower leg and split-thickness skin grafts, the patient was eventually discharged in a stable condition. PMID- 17113525 TI - Bilateral dermal thymus of neck in branchio-oculo-facial syndrome. AB - Dermal thymus in bilateral sides of neck is very rare, probably unique anomaly of branchio-oculo-facial syndrome (BOF). We report a case of BOF with cleft lip and non-healing erosions on bilateral sides of neck. The thymus of neck in our case showed unusual clinical course that appeared at 1 year and half of age after the repair of cleft lip deformity. The lesions of neck were completely excised, and the histopathologic examination for neck lesion confirmed ectopic dermal thymus. Ectopic thymus can be excised completely after careful evaluation of haematologic, immunologic status if normal thymic shadow cannot be identified on chest X-p. PMID- 17113526 TI - Craniofacial cleft: a case of Tessier no. 3, 7 and 11 cleft. AB - The incidence of the rare facial clefts is between 1.43 and 4.85 per 100.000 births. We report a case of right associated Tessier no. 3, 7 and 11 craniofacial clefts with cardiac malformation. The epidemiology, classification, embryology and pathogenesis of each craniofacial malformation are briefly reviewed. After an extensive review of the literature, we conclude that this association has not previously been reported. PMID- 17113527 TI - The first case of primary metacarpal V restoration with titanium mesh and cancellous bone graft. AB - Any references of use of the titanium mesh in hand reconstruction could not be found. A case of primary metacarpal reconstruction after severe hand trauma with a help of cage made of titanium mesh and cancellous iliac bone graft is presented. PMID- 17113528 TI - Recurrent carpal tunnel syndrome in a child due to fibrolipomatous hamartoma of the median nerve successfully treated by limited excision and decompression. AB - We present a case of recurrent carpal tunnel syndrome in a child caused by fibrolipomatous hamartoma of the median nerve which was successfully treated by limited excision of the fibrolipomatous tissue and decompression. PMID- 17113529 TI - Inclusion of a skin strip into Goldstein's myomucosal flap for labial reconstruction. AB - Traumatic defects of the vermilion are often associated with defects of the adjacent skin. We present a case in which we reconstructed a combined labial defect, both mucosal and cutaneous, by means of a composite advancement flap consisting of orbicularis oris muscle, oral mucosa and underlying skin. PMID- 17113530 TI - Are primary cutaneous carcinisarcomas underdiagnosed? Five cases and a review of the literature. AB - Primary cutaneous carcinosarcoma is a biphasic tumour containing both malignant epithelial and malignant mesenchymal elements. To date, only 26 cases have been reported in the literature. However, our findings suggest that this may reflect underreporting and possibly underdiagnosis. We present five cases and a comprehensive review of the literature: The disease most commonly presents in the eighth and ninth decades of life, is twice as common in males and may be related to sun exposure. Surgery is the primary therapeutic modality. Despite treatment, 27% of cases developed metastatic disease. Both epithelial and mesenchymal elements have been implicated in disease spread. We hope to raise awareness of this uncommon but serious disease and to assist clinicians in its management. PMID- 17113531 TI - Complications of polyalkylimide 4% injections (Bio-Alcamid): a report of 18 cases. AB - Injectable filler materials can be valuable to aesthetic surgeons. To date, hardly any short-term and no long-term complications of polyalkylimide injections (Bio-Alcamid) have been reported. We present and discuss the history of 18 patients who had such complications. The patients were between 31 and 55 years of age. The time between injection and the onset of complications of polyalkylimide ranged from 1 month to 3 years. Additional invasive therapy at, or near, the site of injections triggered the onset of infection in 10 patients. By use of T2 weighted MRI with fat suppressing spectro-presaturation inversion recovery (SPIR) the filler material can be visualised. Once infection or migration of the permanent filler occurs, the therapeutic options are limited to surgical removal by a direct approach. Polyalkylimide should be handled under strict antiseptic circumstances. This does not only apply at the time of initial injections, but even more during any subsequent invasive treatment such as evacuation of surplus deposits or additional surgical procedures at, or near, the site of injection. PMID- 17113532 TI - A prepelvic tunnel for the rectus abdominis myocutaneous flap in perineal reconstruction. AB - BACKGROUND: The use of the transpelvic vertical rectus abdominis myocutaneous (VRAM) flap in pelvic reconstruction is well documented. It can be used to fill large defects after pelvic exenteration, reconstruct the vagina and provide skin coverage in perineal reconstruction. This study examines an alternate prepelvic pathway for the flap to enhance its versatility and reliability. PATIENTS AND METHOD: A female patient with recurrent squamous cell carcinoma in the pelvis, who underwent radical pelvic exenteration and a successful VRAM flap reconstruction with a prepelvic tunnel. RESULTS: The patient experienced a small area of epithelial tip necrosis over the sacral promontory from shear forces. This healed with dressings within two weeks. There were no major flap complications and the patient had good flap integrity at one-year follow-up. The prepelvic pathway for the VRAM flap is advantageous to the conventional transpelvic course in perineal reconstruction. The more direct, shorter path to the defect allows for a more reliable skin paddle design without the need for de epithelialisation. A greater area of skin paddle is available and creates a more versatile flap with no tension on the pedicle. This is especially in cases where a skin paddle is needed for vaginal reconstruction or when pelvic organs such as bladder and uterus are left in situ. These advantages may result in less flap complications. PMID- 17113533 TI - Tube pedicle flap in the management of a Grade III C lower limb injury. AB - Salvage of a Grade III C lower limb injury is a challenging problem. Apart from microsurgical revascularisation, they frequently need soft tissue coverage procedures. Due to the magnitude of the injury, local flaps from adjacent tissues may not be available and microsurgical free flaps are the flaps of choice. We present an instance where the defect in the middle third of a revascularised leg was covered by a tube pedicle flap raised from the groin and transferred, with the wrist as the carrier. This was chosen because of the below knee amputation on the opposite side combined with long segment circumferential loss of skin in the affected extremity and paucity of vein graft sources. Thus the reconstruction of a limb which was saved by microsurgery was completed by tube pedicle flap. In exceptional circumstances 'old' techniques are still useful. PMID- 17113534 TI - V-N plasty for the release of severe postburn contractures. AB - The authors describe a technique for the release of severe postburn contractures of the web spaces by using all the available tissues in the web space as local flaps to avoid using skin grafts or distant flaps. We rely on the central part of the web to raise the main triangular flaps that may be used to create a functional web space. We called it the V-N plasty. PMID- 17113535 TI - Reconstruction of abdominal wall by whole thigh flap. AB - Closure of extensive abdominal wall defects can be a very challenging task as there are no known large local or free vascularized flaps available that could cover the entire abdomen. Tensor fascia latae (TFL) has been widely used for abdominal wall reconstruction [Hill HL, Nahai F, Vasocnez LO. The tensor fascia lata myocutaneous free flap. Plast Reconstr Surg 1978;61:517-22]. However, the dimensions of the standard TFL flap limit its use in cases of large full thickness abdominal wall defects. Therefore, we have used an ingenious technique of raising the entire thigh skin as a fasciocutaneous flap (whole thigh flap) based on the concept of fusion of angiosomal territories, to reconstruct such a defect following excision of a large abdominal wall tumour. PMID- 17113536 TI - Preservation of lateral thoracic artery to improve vascular supply of distal skin without compromising pedicle length in harvesting pectoralis major myocutaneous flap. AB - The pectoralis major myocutaneous (PM) flap is supplied by three arterial systems. The lower chest skin of the PM flap is mainly supplied by the branches of lateral thoracic artery and internal mammary artery. The conventional harvesting technique for head and neck reconstruction utilizes single arterial supply from the pectoral branch of thoracoacromial artery. The distal skin island of PM flap is therefore compromised and requires indirect blood supply by communicating vessels. In harvesting the PM flap, the pectoralis minor muscle is divided to preserve the lateral thoracic artery and its blood supply to the lateral distal skin island of PM flap without compromising the pedicle length for head and neck reconstruction. Six PM flaps were harvested for reconstruction of head and neck defects with preservation of both the pectoral artery and lateral thoracic artery. The focal pint of swing of all six flaps was at the same point just below the mid-point of clavicle for both pectoral artery and lateral thoracic artery. The flaps can reach the oral cavity, tonsil or hypopharynx without limitation and there is no flap necrosis. In conclusion, the lateral thoracic artery can be preserved without compromising the pedicle length of PM flap. It is a recommended technique to improve the blood supply to the distal skin of PM flap. PMID- 17113537 TI - A new flap design for reconstruction of full-thickness defects of the lower lip: the extended upper lip island (EULI) flap. AB - The extended upper lip island (EULI) flap has two advantages. It provides a wide mucosal surface, like that of the facial artery musculomucosal flap and does not leave a pedicle across the mouth. Aesthetic results with the EULI flap are almost as good as those with the cross-lip flap. An 89-year-old woman presented with squamous cell carcinoma of the lower oral vestibule. Full-thickness lower lip resection and marginal mandibulectomy, including resection of the surrounding gingiva and mucosa, were performed. The defect was reconstructed with an EULI flap that included the facial artery and vein. There were no severe postoperative complications, but two mild complications did occur: mild congestion of the distal end of the skin paddle and mucosa for two days after surgery and partial paralysis of the orbicularis oris and levator anguli oris muscles. The EULI flap is useful for extended lower lip reconstruction. PMID- 17113538 TI - A review of the surgical treatment of vulval lymphangioma and lymphangiectasia: four case reviews. AB - OBJECTIVE: To evaluate whether surgical management of vulval lymphoedema and/or lymphangiectasia conveys any longstanding patient benefit. PATIENT AND METHODS: A qualitative analysis of signs and symptoms that occurred before and after surgical treatment for vulval lymphoedema and/or lymphangiectasia was performed. This was done by analysis of patient notes and telephone conducted interview. RESULTS: From a hospital database search, four patients were found to have had surgical treatment--three for lymphangioma and one for lymphangiectasia. Overall there was a clear improvement in the signs and symptoms associated with these conditions. In particular, all patients reported an improvement (i.e. a reduction or elimination) in the amount of oedema following surgery. CONCLUSION: Carbon dioxide laser therapy and superficial radiotherapy have been previously described for the management of vulval lymphangioma and lymphangiectasia with limited success, whereas our data suggests surgery offers a more permanent solution. In particular, labial reduction seems to be more successful than methods such as lymphovenous anastomoses and lymphangioplasties. A single operation may provide benefit for up to ten years. This approach has the potential to allow patients to be rehabilitated to normal life and activity. PMID- 17113539 TI - Operating in an eczematous surgical field: don't be rash, delay surgery to avoid infective complications. AB - Prominent ear correction is a common operation. Complication as a result of infection has been quoted at between 3% and 5% [Calder JC, Nasaan A. Morbidity of otoplasty: a review of 562 consecutive cases. Br J Plast Surg 1994;47:170-4 and Jeffery SLA. Complications following correction of prominent ears: an audit review of 122 cases. Br J Plast Surg 1999;52:588-90.]. We present two cases referred for ear reconstruction following catastrophic post-operative infection at the time of pinnaplasty, leaving each patient with significant helical rim deformities. Both patients displayed evidence of active post-auricular eczema at the time of their primary surgery. Dermatological research has highlighted the increased colonisation of Staphylococcus aureus in particular within areas of atopic eczema in comparison to normal skin. We advise delaying ear surgery in the presence of a rash in view of the potentially devastating complications that may result. This approach may be extended to all cutaneous surgery where treatment of the rash is advocated prior to embarking on an elective surgical procedure. PMID- 17113540 TI - Treatment of auricular pseudocyst with aspiration and local pressure. AB - An auricular pseudocyst is a fluid filled cavity unlined by epithelium in the intracartilaginous space of the ear. Clinically, it presents as a painless outpouching on the upper anterior surface of the auricle with sterile straw coloured fluid found upon aspiration. If left untreated, permanent deformity of the pinna may result. Treatment of the pseudocysts has varied in the literature with techniques involving aspiration alone or incision and drainage; however, due to the high recurrence rates with these methods more aggressive therapies have been instituted. Although these techniques have achieved reasonable results with few recurrences, their invasive nature and chemotherapeutic and toxic agents used have caused significant morbidity. We present treatment of an auricular pseudocyst by simple aspiration and local pressure application using an auricular prosthesis formulated with the creation of a moulage fitted to the ear by our prosthetist. This minimally invasive and simplistic approach can avoid some of the complications related to other therapies, such as cartilage deformity, depigmentation of skin, and scarring and yields optimal cosmetic results. PMID- 17113541 TI - Surgical approach to the congenital megaprepuce. AB - The megaprepuce (MP) is a very rare malformation of unknown aetiology, with anatomical findings similar to those observed in the congenital buried penis (BP). The aspect of this entity and the symptoms are rather typical: the penis is totally buried and before micturition an evident pubic and scrotal swelling denounces the urine collection in an abnormal preputial cavity surrounding the entire shaft. Early surgery is recommended, in order to deal with functional and cosmetic aspects. The very diminutive phallus is the most important cause of family anxiety. The authors present six infants, aged 5, 12, 13, 18, 20 and 43 months, operated upon for MP. The surgical approach consisted in the complete exteriorization of the shaft with section of the penile ligament, the restoring of the pubo-penile and peno-scrotal angles and the tailoring of the cutaneous cylinder. No intra- or postoperative complications were observed. Late functional and cosmetic results were judged satisfactory by both parents and surgeons. The surgical approach to BP is not simply transferable to the correction of MP. The cosmetic arrangement of the shaft skin is the more difficult step of the MP correction in order to avoid postoperative complications and an unsatisfied cosmetic appearance. PMID- 17113542 TI - Malignant peripheral nerve sheath tumour of the cervical vagus nerve in a neurofibromatosis type 1 patient. AB - One serious complication of neurofibromatosis type 1 (NF1) is the development of malignant peripheral nerve sheath tumours (MPNSTs). These malignancies often develop within pre-existing plexiform neurofibromas and their development is now thought to be associated with both tumour suppressor gene mutations and dysregulated growth factor signalling. Recent work demonstrates that the lifetime risk of malignant transformation is significantly greater than previously thought. Ionising radiation, a long-standing disease, particularly the presence of a large number of plexiform neurofibromas from an early age, are suggested risk factors. We present an NF1 patient who developed an MPNST of the cervical vagus nerve which was successfully treated with surgery. Close monitoring of patients with NF and a high level of suspicion towards rapidly enlarging and painful swellings is merited as these features may signify malignant transformation. Whether a positive history of MPNST in other affected family members predisposes the individual to a higher risk of malignant transformation is unclear. PMID- 17113543 TI - Three episodes of gracilis free muscle transfer under epidural anaesthesia. AB - The use of regional anaesthesia in major surgery is associated with a lower risk of complications. However, recent evidence suggests that a vascular steal phenomenon may result in a reduction of free flap blood flow in such patients. We report three cases of free gracilis transfer under epidural anaesthesia in patients who were considered high risk for general anaesthesia. Our experience suggests that there remains an important role for epidural anaesthesia in the management of patients undergoing lower limb free flap reconstruction. The inability to undergo general anaesthesia does not preclude free flap surgery in carefully selected patients. PMID- 17113544 TI - Lack of operative experience is only the tip of the iceberg for plastic surgery SHOs in the UK, in response to Wong et al. [The effect of the new deal on the operative experience of plastic surgical SHOs' (Journal of Plastic, Reconstructive and Aesthetic Surgery, Volume 59, Issue 3, March 2006, Pages 311 312)]. PMID- 17113545 TI - Correction of long term joint contractures of the hand by distraction. A case report. PMID- 17113546 TI - Comment on "An adherent dressing for aplasia cutis congenita". PMID- 17113547 TI - Reply to 'Raghavan U, Jones NS. The complications of giant titanium implants in nasal reconstruction. J Plast Reconstr Aesthetic Surg (2006);59:74-79'. PMID- 17113548 TI - In response to 'Facial herpes simplex infection from possible cross contamination through the laser handpiece following cutaneous laser resurfacing'. PMID- 17113549 TI - Nipple sharing for nipple-areolar complex reconstruction: still a useful technique. PMID- 17113550 TI - Modernizing medical careers in the UK and plastic surgery as a possible career choice: undergraduate opinions. PMID- 17113551 TI - A laser pointer guide to take accurate intraoperative photographs. PMID- 17113552 TI - A crash-prediction model for multilane roads. AB - Considerable research has been carried out in recent years to establish relationships between crashes and traffic flow, geometric infrastructure characteristics and environmental factors for two-lane rural roads. Crash prediction models focused on multilane rural roads, however, have rarely been investigated. In addition, most research has paid but little attention to the safety effects of variables such as stopping sight distance and pavement surface characteristics. Moreover, the statistical approaches have generally included Poisson and Negative Binomial regression models, whilst Negative Multinomial regression model has been used to a lesser extent. Finally, as far as the authors are aware, prediction models involving all the above-mentioned factors have still not been developed in Italy for multilane roads, such as motorways. Thus, in this paper crash-prediction models for a four-lane median-divided Italian motorway were set up on the basis of accident data observed during a 5-year monitoring period extending between 1999 and 2003. The Poisson, Negative Binomial and Negative Multinomial regression models, applied separately to tangents and curves, were used to model the frequency of accident occurrence. Model parameters were estimated by the Maximum Likelihood Method, and the Generalized Likelihood Ratio Test was applied to detect the significant variables to be included in the model equation. Goodness-of-fit was measured by means of both the explained fraction of total variation and the explained fraction of systematic variation. The Cumulative Residuals Method was also used to test the adequacy of a regression model throughout the range of each variable. The candidate set of explanatory variables was: length (L), curvature (1/R), annual average daily traffic (AADT), sight distance (SD), side friction coefficient (SFC), longitudinal slope (LS) and the presence of a junction (J). Separate prediction models for total crashes and for fatal and injury crashes only were considered. For curves it is shown that significant variables are L, 1/R and AADT, whereas for tangents they are L, AADT and junctions. The effect of rain precipitation was analysed on the basis of hourly rainfall data and assumptions about drying time. It is shown that a wet pavement significantly increases the number of crashes. The models developed in this paper for Italian motorways appear to be useful for many applications such as the detection of critical factors, the estimation of accident reduction due to infrastructure and pavement improvement, and the predictions of accidents counts when comparing different design options. Thus this research may represent a point of reference for engineers in adjusting or designing multilane roads. PMID- 17113553 TI - Increased expression of iron-containing superoxide dismutase-A (TcFeSOD-A) enzyme in Trypanosoma cruzi population with in vitro-induced resistance to benznidazole. AB - Superoxide dismutase (SOD) removes excess superoxide radicals via dismutation to oxygen and hydrogen peroxide. In this work, we have characterized TcFeSOD-A gene from 25 Trypanosoma cruzi populations and clones susceptible, naturally resistant or with in vitro-induced (17 LER) or in vivo-selected resistance to benznidazole (BZR). In the 17 LER T. cruzi population, the levels of TcFeSOD-A mRNA were at least 3-fold higher than its drug-susceptible counterpart 17 WTS. The levels of TcFeSOD-A mRNA were similar among the other T. cruzi populations and clones regardless of the drug-resistance phenotype. We determined whether the increase in mRNA levels was due to gene amplification using Southern blot analysis of the T. cruzi populations and clones. We found that the number of TcFeSOD-A gene copies was similar for all samples tested, except for 17 LER that presented twice as many copies. The chromosomal location of the TcFeSOD-A gene and polymorphisms detected in nucleotide and amino acid sequences of TcFeSOD-A were associated with the zymodeme of the T. cruzi strain but not with drug-resistance phenotype. We observed a 23 kDa TcFeSOD-A polypeptide in all analysed T. cruzi strains. The level of this polypeptide was increased only in the 17 LER population. Specific enzyme activity analysis of TcFeSOD in the T. cruzi samples revealed a correlation between expression and activity. Our findings show an increased expression of the TcFeSOD-A enzyme in the T. cruzi population with in vitro induced resistance to benznidazole. PMID- 17113554 TI - Improved efficacy with amodiaquine instead of chloroquine in sulfadoxine/pyrimethamine combination treatment of falciparum malaria in Uganda: experience with fixed-dose formulation. AB - Amodiaquine (AQ) is an affordable compound, chemically related to chloroquine (CQ) but often effective against CQ resistant Plasmodium falciparum. In Uganda, a pre-packed fixed-dose combination of CQ plus sulfadoxine/pyrimethamine (CQ+SP) called Homapak is used in the home based management of fever program (HBM). We performed a single blind randomized trial to determine the efficacy of AQ+SP in comparison with the fixed-dose CQ+SP (Homapak) in the treatment of uncomplicated falciparum malaria in Ugandan children aged 6 months to 5 years. The study was done in 2004 at Walkuba Health Center, a sub-urban area in Jinja district, Uganda. Primary outcome was the day 14 per protocol clinical and parasitological response according to the WHO. A total of 183 children were included (mean age 28 months) and 90% completed 28 days of follow up. The day 14 adequate clinical and parasitological response was 70.9% for CQ+SP and 97.4% for AQ+SP (p<0.001). In those given CQ+SP, treatment failure rates for the 6 months to 2 years age group were much higher (48.2%) than in the older children (18.2%, p=0.004). The day 28 PCR adjusted parasitological failure rates were also higher in the CQ+SP (31.3%) than in the AQ+SP group (13.1%) (p=0.003), with a higher gametocyte carriage among the CQ+SP group. We conclude that the efficacy of AQ+SP was significantly superior to the fixed-dose CQ+SP (Homapak), particularly among the youngest children. Thus, AQ could be used instead of CQ in combination with SP to improve the effectiveness against falciparum malaria in Uganda. PMID- 17113555 TI - Demographic and spatio-temporal variation in human plague at a persistent focus in Tanzania. AB - Human plague in the Western Usambara Mountains in Tanzania has been a public health problem since the first outbreak in 1980. The wildlife reservoir is unknown and eradication measures that have proved effective elsewhere in Tanzania appear to fail in this region. We use census data from 2002 and hospital records kept since 1986 to describe the temporal, spatial and demographic variation in human plague. A seasonal peak in cases occurs from December to February with the numbers of cases during this peak varying between 0 and 1150. Variation in incidence, calculated for each village as the mean number of cases per thousand inhabitants per year, indicates that human plague is concentrated around a group of three neighbouring, relatively isolated, high-altitude villages; Nywelo, Madala and Gologolo. However, there was no evidence that these villages were acting as a source of infection for the remainder of the focus. The likelihood of becoming infected with plague is highest between the ages of 5 and 19 and lowest for adult men. This was most clear in the ward encompassing the three high incidence villages where the risk of plague among children aged 10-14 was 2.2 times higher than for adults aged 30-34, and among adults aged 30-34, the risk was 2.4 times higher for women than men. PMID- 17113556 TI - Anisometropic amblyopia treated with spectacle correction alone: possible factors predicting success and time to start patching. AB - PURPOSE: To evaluate factors predicting resolution of amblyopia and the time course of improvement in VA in children 3 to 7 years old with anisometropic amblyopia treated with spectacles alone. DESIGN: Prospective, noncomparative intervention. METHODS: Measurement of corrected amblyopic logMAR visual acuity (VA) in newly diagnosed children at four-weekly intervals until VA stabilized or amblyopia resolved. The time course of improvement in VA and the factors related to amblyopia resolution were assessed. RESULTS: Sixty children with a mean age of 5.3 years and mean anisometropia of 2.95 diopters (D) were included. Amblyopia improved by 2 or more logMAR lines in 56 patients (93%) and resolved in 27 patients (45%), with a mean improvement in VA of 0.38 logMAR. The improvement in VA in the amblyopic eye was considerable at four to 12 weeks then reached a plateau, after which it improved only slowly. Resolution of amblyopia was related to better initial VA (0.2 to 0.6 logMAR) and lesser amounts of anisometropia (<4 D). The time to resolution ranged from four to 40 weeks, but no patient with residual amblyopia showed an improvement in VA of more than 0.1 logMAR over four consecutive visits. CONCLUSIONS: With spectacle correction alone, 3- to 7-year old children with previously untreated anisometropic amblyopia achieved approximately four-line improvement and resolved nearly in half. The nearly two month plateau periods during improvement of VA should be noticed. After four months with no improvement in VA, occlusion therapy or atropine penalization may be considered. PMID- 17113557 TI - Radial optic neurotomy in central retinal vein occlusion: comparison of outcome in younger vs older patients. AB - PURPOSE: To evaluate the effectiveness of radial optic neurotomy (RON) for central retinal vein occlusion (CRVO) in patients < or =50 years of age (group 1) vs those >50 (group 2). DESIGN: Prospective, interventional, comparative case series. METHODS: The study included 43 consecutive patients with CRVO and preoperative visual acuity (VA) < or =0.70 logarithm of minimal angle of resolution (logMAR). All patients underwent pars plana vitrectomy and RON at the nasal border of the optic disk. VA and optical coherence tomography (OCT) findings were recorded preoperatively and at one, six, and 12 months postoperatively. RESULTS: Systemic hypertension, diabetes, and open-angle glaucoma were statistically significantly more prevalent in group 2 (P < .05). One patient in group 1 had hyperhomocysteinemia, and had another antiphospholipid syndrome. Fifty percent of patients in group 1 gained > or =3 lines of Early Treatment Diabetic Retinopathy Study (ETDRS) vision, vs 33% in group 2. Mean final VA was 0.5 logMAR VA in group 1 vs 0.8 in group 2 (P = .04). Foveal thickness decreased significantly in both groups (P < .001). Ten patients (55.6%) in group 1 and 13 (54.2%) in group 2 developed retinochoroidal collaterals. CONCLUSIONS: Underlying systemic disease does not seem to be an important factor in the pathogenesis of CRVO in younger patients, and thrombophilia was present in only 11% of patients in this age group. RON yielded better functional results in younger patients, although functional improvement remained limited in those with low baseline VA. PMID- 17113558 TI - Binding thermodynamics of substituted diaminopyrimidine renin inhibitors. AB - Renin is an aspartyl protease involved in the production of angiotensin II, a potent vasoconstrictor. Renin inhibitors can prevent blood vessel constriction and therefore could be useful for the treatment of hypertension. High-throughput screening efforts identified a small molecule renin inhibitor with a core substituted diaminopyrimidine ring. Parallel medicinal chemistry efforts based on this lead resulted in compound 1. A complex of 1 bound to renin was crystallized, and structural data were obtained by X-ray diffraction. The structure indicated that there were adjacent unoccupied binding pockets. Synthetic efforts were initiated to extend functionality into these pockets so as to improve affinity and adjust pharmacokinetic parameters. Thermodynamics data for inhibitor binding to renin were also collected using isothermal titration calorimetry. These data were used to help guide inhibitor optimization by suggesting molecular alterations to improve binding affinity from both thermodynamic and structural perspectives. The addition of a methoxypropyl group extending into the S3 subpocket improved inhibitor affinity and resulted in greater binding enthalpy. Initial additions to the pyrimidine ring template that extended into the large hydrophobic S2 pocket did not improve affinity and dramatically altered the thermodynamic driving force for the binding interaction. Binding of the core template was enthalpically driven, whereas binding of initial inhibitors with S2 extensions was both enthalpically and entropically driven but lost significant binding enthalpy. Additional electrostatic interactions were then incorporated into the S2 extension to improve binding enthalpy while taking advantage of the favorable entropy. PMID- 17113559 TI - Development and specific induction of apoptosis of cultured cell models overexpressing human tau during neural differentiation: Implication in Alzheimer's disease. AB - Apoptosis or programmed cell death is considered to be involved in neurodegenerative disorders, including Alzheimer's disease (AD). AD is characterized by intracellular aggregates of hyperphosphorylated tau, a microtubule-associated protein. To investigate the induction of apoptosis by abnormal tau resembling AD, cultured cells may be useful tools. We developed a cell culture model and established NG108-15 and P19 cells stably transfected with human tau, naming them tau/NG and tau/P19 cells, respectively. Increased accumulation and phosphorylation of tau were observed during neural differentiation in tau/NG cells. Tau/P19 cells underwent drastic apoptosis during neural differentiation induced by retinoic acid (RA). Tau protein was distributed throughout the cytoplasm and in specific zones of the nucleus. The cytoplasmic tau was associated with microtubules, but the nucleic tau was observed to form clusters and was associated with RA receptor (RAR). The apoptosis induced by RA was inhibited by the treatment of glycogen synthase kinase 3 (GSK3) inhibitor in tau/P19 cells. We propose that translocation of tau into nucleus affects RA signaling in apoptosis via GSK3 in the cells. These cells are useful for monitoring the apoptosis by abundant tau and may be applied to investigate the molecular mechanism of apoptosis resembling AD. PMID- 17113560 TI - One-step generation of degraded DNA by UV irradiation. PMID- 17113561 TI - Fiber-based single cell analysis of reporter gene expression in yeast two-hybrid systems. AB - Yeast two-hybrid (Y2H) systems conventionally are used when screening for protein protein interactions. A recurring problem with Y2H systems is the high prevalence of both false positives and false negatives. Distinguishing between real results and artifacts to characterize a protein-protein interaction properly can often be time-consuming and laborious. In this article, we report a new method for statistically evaluating single cell reporter gene transcriptional activation and expression in yeast two-hybrid systems. The system is based on statistical analysis of many yeast cells contained in a fiber-optic array, allowing for thorough characterization of reporter gene expression at different stringency levels. PMID- 17113562 TI - Genotyping of eight polymorphic genes encoding drug-metabolizing enzymes and transporters using a customized oligonucleotide array. AB - Polymorphisms in drug-metabolizing genes may lead to the production of dysfunctional proteins and consequently affect therapeutic efficacy and toxicity of drugs. Different frequencies of polymorphic alleles among the races have been postulated to account for the observed ethnic variations in drug responses. In the current study, we aimed to estimate the frequencies of 14 polymorphisms in eight genes (TPMT, NQO1, MTHFR, GSTP1, CYP1A1, CYP2D6, ABCB1, and SLC19A1) in the Singapore multiracial populations by screening 371 cord blood samples from healthy newborns. To improve genotyping efficacy, we designed an oligonucleotide array based on the principle of allele-specific primer extension (AsPEX) that was capable of detecting the 14 polymorphisms simultaneously. Cross-validation using conventional polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) assays demonstrated 99% concordant results. Measurements on the fluorescent intensity displayed clear distinctions among different genotypes. Statistical analyses showed significantly different allele distributions in several genes among the three races, namely Chinese, Malays, and Indians. Comparing the allelic frequencies in Chinese with previous studies in Caucasian populations, NQO1 609C>T and SLC19A1 80G>A were distinctly different, whereas close similarity was observed for MTHFR 677C>T. We have demonstrated an array based methodology for rapid multiplex detection of genetic polymorphisms. The allelic frequencies reported in this study may have important therapeutic and prognostic implications in the clinical use of relevant drugs. PMID- 17113563 TI - Cation exchange-HPLC and mass spectrometry reveal C-terminal amidation of an IgG1 heavy chain. AB - A unique, late-eluting "basic peak" (relative to the "main peak") was observed by weak cation exchange-HPLC (WCX) for a recombinant monoclonal antibody (mAb) sample. Peak fractions were collected, desalted, and analyzed by high-resolution MS using a top-down characterization approach that provided accurate masses of intact mAb charge isoforms and a comprehensive profile of the structural heterogeneity. The individual light (L) and heavy (H) chain subunits from the main and basic peaks were analyzed by reversed-phase (RP) HPLC/MS after disulfide bond reduction and cysteine alkylation. Three mAb isoforms were detected, and their modifications were localized to H chain. Bottom-up characterization using RP-HPLC/MS peptide mapping and accurate mass measurements identified three distinct H chain C-terminal peptides ending in glycine, lysine, or alpha-amidated proline. The combined analyses showed that the main WCX peak mAb isoform contained two unmodified L chains and two H chains terminating in glycine. Each mAb isoform that coeluted in the basic peak consisted of two unmodified L chain subunits and a single H chain ending in glycine, but the second H chain terminated in lysine for one isoform and alpha-amidated proline for another isoform. The WCX elution positions of the isoforms were consistent with their respective net charge. To the best of our knowledge, the occurrence of C-terminal alpha-amidation in mAbs has not been reported previously. PMID- 17113564 TI - Comparison of stoop versus prone postures for a simulated agricultural harvesting task. AB - Physical and psychophysical differences between working in the stooped and prone postures were compared while performing a simulated agricultural harvesting task for 30 min. Fifteen male subjects participated. The measures used to compare the two postures included perceived discomfort, electromyography (EMG), and heart rate (HR). Average hamstrings localized discomfort (0-10 scale) was 6.17 (SD=2.9) for the stoop posture and 0.67 (SD=1.29) for the prone posture. Erector spinae and hamstring EMG RMS increased 68% and 18%, respectively, while mean power frequency for the hamstrings decreased 13% for the stoop task. Mean power frequency for the middle trapezius muscle decreased in both postures (stoop 4.13%, prone 3.79%). Average heart rate during the last work cycle was 35% greater than the resting heart rate for the stoop posture while average heart rate was 17% greater for the prone posture. Subjects worked on the prone workstation without rest during the 15 min work simulations with less discomfort, no localized fatigue in the back or leg muscles tested, and lower working heart rates than subjects working in the stoop posture. PMID- 17113565 TI - Roles of basic residues and salt-bridge interaction in a vacuolar H+-pumping pyrophosphatase (AVP1) from Arabidopsis thaliana. AB - To investigate the possible role of basic residues in H+ translocation through vacuolar-type H+-pumping pyrophosphatases (V-PPases), conserved arginine and lysine residues predicted to reside within or close to transmembrane domains of an Arabidopsis thaliana V-PPase (AVP1) were subjected to site-directed mutagenesis. One of these mutants (K461A) exhibited a "decoupled" phenotype in which proton-pumping but not hydrolysis was inhibited. Similar results were reported previously for an E427Q mutant, resulting in the proposal that E427 might be involved in proton translocation. However, the double mutant E427K/K461E has a wild type phenotype, suggesting that E427 and K461 form a stabilising salt bridge, but that neither residue plays a critical role in proton translocation. PMID- 17113566 TI - Detection of emotional expressions in rapidly changing facial displays in high- and low-socially anxious women. AB - Facial information and attention to facial displays are distributed over spatial as well as temporal domains. Thus far, research on selective attention to (dis)approving faces in the context of social anxiety has concentrated primarily on the spatial domain. Using a rapid serial visual presentation (RSVP) paradigm, the present study examined the temporal characteristics of visual attention for happy and angry faces in high- (n=16) and low-socially anxious individuals (n=17), to test whether also in the temporal domain socially anxious individuals are characterized by threat-confirming attentional biases. Results indicated that presenting angry faces as the first target (T1) did not aggravate the detection of the emotional expression of the second target (T2). Yet, participants generally showed superior detection of the emotional expression of T2, if T2 was an angry face. Casting doubt on the role of such attenuated attentional blink for angry faces in social anxiety, no evidence emerged to indicate that this effect was relatively strong in high-socially anxious individuals. Finally, the presentation of an angry face as T2 resulted in a relatively hampered identification of a happy-T1. Again, this "backward blink" was not especially pronounced in high-socially anxious individuals. The present anger superiority effects are consistent with evolutionary models stressing the importance of being especially vigilant for signals of dominance. Since the effects were not especially pronounced in high-anxious individuals, the present study adds to previous findings indicating that socially anxious individuals are not characterized by a bias in the (explicit) detection of emotional expressions [Philippot, P., & Douilliez, C. (2005). Social phobics do not misinterpret facial expression of emotion. Behaviour Research and Therapy, 43, 639-652]. PMID- 17113567 TI - Calcineurin interacts with KIN-29, a Ser/Thr kinase, in Caenorhabditis elegans. AB - Calcineurin is a Ca2+/Calmodulin activated Ser/Thr phosphatase that is well conserved from yeast to human. In Caenorhabditis elegans, tax-6 and cnb-1 encode catalytic and regulatory subunits of calcineurin, respectively. We performed yeast two-hybrid screening using TAX-6 as a bait to identify calcineurin interacting proteins. KIN-29 is one of proteins that specifically interacted with TAX-6. KIN-29 is a Ser/Thr kinase previously shown to be involved in regulating gene expression of a subset of chemoreceptors in specific neurons. Both TAX-6 and KIN-29 are expressed in hypodermis, muscles, and neurons. Moreover, both calcineurin and kin-29 mutants exhibit similar phenotypes, namely small body size, small brood size, and slow growth. Here we describe specific genetic interaction between tax-6 and kin-29 in regulating body size, serotonin mediated egg laying, and chemoreceptor expression. PMID- 17113568 TI - Substrate-dependent inhibition or stimulation of HIV RNase H activity by non nucleoside reverse transcriptase inhibitors (NNRTIs). AB - HIV reverse transcriptase (HIV-RT) contains two distinct protein domains catalyzing DNA polymerase and RNase H activities. Non-nucleoside reverse transcriptase inhibitor (NNRTI) binding to HIV-RT can affect RNase H activity. The structurally diverse NNRTIs capravirine, efavirenz, GW8248, TMC-125, and nevirapine all inhibited 5'-RNA directed HIV RNase H activity as partial inhibitors with maximal inhibition of 40-65%. Potencies of RNase H inhibition correlated with the respective potencies of DNA polymerase inhibition. Mutations in the NNRTI binding site (K103N, Y181C, Y188L, and K103N/Y181C) reduced the potency of RNase H inhibition, similar to their effects on DNA polymerase activity. The NNRTIs did not affect the activity of the isolated HIV RNase H domain. In contrast, 3'-DNA directed RNase H activity of HIV-RT was mechanistically distinct from 5'-RNA directed RNase H activity and was stimulated rather than inhibited by NNRTI binding to HIV-RT. Therefore, NNRTI binding to the polymerase domain of HIV-RT interferes with RNase H activity through a long-range effect, which is affected by the structure of the RNA:DNA hybrid substrate, but is independent of NNRTI compound structure and nucleic acid substrate sequence. PMID- 17113569 TI - Characterization of Schistosoma mansoni ATPDase2 gene, a novel apyrase family member. AB - Schistosoma mansoni is a major causative agent of schistosomiasis, which constitutes a severe health problem in developing countries. We have previously described the SmATPDase1 gene, encoding a protein from the external surface of the parasites. In this work, we describe the cloning and characterization of SmATPDase2, a novel CD39-like ATP diphosphohydrolase gene in S. mansoni. In silico analysis of the protein encoded by SmATPDase2 predicts a single N-terminal transmembrane domain similar to that described for secreted human apyrase isoforms. Immuno-colocalization experiments detected both SmATPDase proteins at the S. mansoni adult worm tegument basal and apical membranes, but only SmATPDase2 in the tegument syncytium. SmATPDase2 but not SmATPDase1 protein was detected by Western blot in culture medium supernatants following incubation of adult worms in vitro, indicating that SmATPDase2 was secreted by the parasite to the medium. Taken together these data suggest a non-redundant role for SmATPDase2 in the parasite-host interplay. PMID- 17113570 TI - Temperature regulation is compromised in experimental limbic status epilepticus. AB - Temperature dysregulation is well known in generalized convulsive status epilepticus but so far has not been reported in non-convulsive forms. In order to detect possible subtle alterations, we have analyzed the capability to compensate for external cooling in an animal model of limbic status epilepticus. Rats with electrically induced self-sustaining status epilepticus (SSSE) (n=6) as well as rats without electrical stimulation (n=6) were cooled for 3 h and then rewarmed for another hour. The time course of changes in epidural temperature in animals of both groups that underwent cooling and in control rats that were not cooled and not stimulated (n=6) was compared. In animals with limbic SSSE, temperature fell continuously and was significantly lower at all time points under cooling as compared with each of the two other groups. In animals that were not stimulated, temperature under cooling fell by 1 to 2 degrees C only and was not significantly different at any time point as compared with controls. The effect of cooling was reversible in both groups. The current data indicate that temperature homeostasis in limbic status epilepticus is markedly disturbed. This finding may suggest ictal involvement of primary thermoregulatory neurons in the anterior hypothalamus probably by spread of epileptic activity from temporo-mesial structures. PMID- 17113571 TI - ERP measures of auditory word repetition and translation priming in bilinguals. AB - Motivated by the demonstration of similarly localized adaptation of the hemodynamic response in a first (L1) and second (L2) language, this study examined event-related brain potentials (ERPs) to spoken words in L1 and L2 in 15 English-French bilinguals. We examined whether the temporal pattern of N400 adaptation due to within-language repetitions (i.e., repetition priming) was similar in L1 and L2 and whether the release from adaptation elicited by a within language word change was similar. Furthermore, using word changes across language, we examined the phonological mismatch negativity (PMN) and N400 components to determine the kind of information activated during translation priming. In contrast to within-language repetition, we expected between-language repetition (i.e., translations) to be characterized by conceptual rather than lexical/phonological word form priming. Overall, the pattern of adaptation and release from adaptation was similar in L1 and L2, with evidence of delayed semantic analysis in L2 in the form of a later N400 effect. A change in language (L1 to L2) elicited a similar pattern of PMN and N400 activity compared to a within-language change in meaning in L1, suggesting that neither word form nor conceptual information was available on-line for the forward translation. In contrast, the presence of strong PMN but minimal N400 effects for L2-to-L1 translations suggests that conceptual but not phonological information is available on-line for backwards translation. L2 proficiency influenced the extent to which conceptual representations were activated by translations. These data are discussed in light of current models of bilingual word processing and suggest modality differences in the pattern of activation of lexical and conceptual information. PMID- 17113572 TI - Effects of time pressure on verbal self-monitoring: an ERP study. AB - The Error-Related Negativity (ERN) is a component of the event-related brain potential (ERP) that is associated with action monitoring and error detection. The present study addressed the question whether or not an ERN occurs after verbal error detection, e.g., during phoneme monitoring. We obtained an ERN following verbal errors which showed a typical decrease in amplitude under severe time pressure. This result demonstrates that the functioning of the verbal self monitoring system is comparable to other performance monitoring, such as action monitoring. Furthermore, we found that participants made more errors in phoneme monitoring under time pressure than in a control condition. This may suggest that time pressure decreases the amount of resources available to a capacity-limited self-monitor thereby leading to more errors. PMID- 17113573 TI - Facilitation of performance in a working memory task with rTMS stimulation of the precuneus: frequency- and time-dependent effects. AB - Although improvements in performance due to TMS have been demonstrated with some cognitive tasks, performance improvement has not previously been demonstrated with working memory tasks. In the present study, a delayed match-to-sample task was used in which repetitive TMS (rTMS) at 1, 5, or 20 Hz was applied to either left dorsolateral prefrontal or midline parietal cortex during the retention (delay) phase of the task. Only 5 Hz stimulation to the parietal site resulted in a significant decrease in reaction time (RT) without a corresponding decrease in accuracy. This finding was replicated in a second experiment, in which 5 Hz rTMS at the parietal site was applied during the retention phase or during presentation of the recognition probe. Significant speeding of RT occurred in the retention phase but not the probe phase. This finding suggests that TMS may improve working memory performance, in a manner that is specific to the timing of stimulation relative to performance of the task, and to stimulation frequency. PMID- 17113574 TI - Endothelial nitric oxide synthase activation leads to dilatory H2O2 production in mouse cerebral arteries. AB - OBJECTIVE: Hydrogen peroxide (H2O2) produced by the vascular endothelium is a signaling molecule regulating vascular tone. We hypothesized that H2O2 derived from eNOS activity could play a physiological role in endothelium-dependent dilation of mouse cerebral arteries. METHODS: Simultaneous endothelium-dependent dilation and fluorescence-associated free radical (DCF-DA) or NO (DAF-2) production were recorded in isolated and pressurized (60 mm Hg) cerebral artery of C57Bl/6 male mice. RESULTS: Without synergism, N-nitro-L-arginine (L-NNA) or the H2O2 scavengers catalase, PEG-catalase and pyruvate reduced (P < 0.05) by 50% the endothelium-dependent dilation induced by acetylcholine (ACh). Simultaneously with the dilation, H2O2--but not NO--production, sensitive to either L-NNA or catalase, was detected. In cerebral arteries from C57Bl/6.eNOS-/- mice, catalase had no effect on ACh-induced dilation and no H2O2-associated fluorescence was observed. In C57Bl/6 mice, silver diethyldithiocarbamate (DETC), a superoxide dismutase (SOD) inhibitor, but not the specific NO scavenger 2-phenyl-4,4,5,5 tetramethyl-imidazoline-1-oxyl3-oxide (PTIO), prevented ACh-induced dilation and H2O2 production suggesting that eNOS-derived superoxide is an intermediate in the production of H2O2. The catalase-sensitive ACh-induced dilation was restored by the eNOS cofactor tetrahydrobiopterin (BH4). This reversal was associated with a NO-associated fluorescence sensitive to PTIO but not to catalase. Soluble guanylate cyclase inhibition with 1H-[1,2,4]-oxadiazole-4,3-aquinoxalin-1-one (ODQ) prevented the dilation induced by ACh and by exogenous H2O2. Lastly, L-NNA, PTIO and ODQ--but not DETC, catalase or pyruvate--increased the pressure dependent myogenic tone, suggesting that eNOS produces NO at rest, but leads to H2O2 during muscarinic stimulation. CONCLUSION: H2O2-dependent dilation in mouse cerebral arteries appears to be a physiological eNOS-derived mechanism. PMID- 17113575 TI - Expression of Hoxa2 in rhombomere 4 is regulated by a conserved cross-regulatory mechanism dependent upon Hoxb1. AB - The Hoxa2 gene is an important component of regulatory events during hindbrain segmentation and head development in vertebrates. In this study we have used sequenced comparisons of the Hoxa2 locus from 12 vertebrate species in combination with detailed regulatory analyses in mouse and chicken embryos to characterize the mechanistic basis for the regulation of Hoxa2 in rhombomere (r) 4. A highly conserved region in the Hoxa2 intron functions as an r4 enhancer. In vitro binding studies demonstrate that within the conserved region three bipartite Hox/Pbx binding sites (PH1-PH3) in combination with a single binding site for Pbx-Prep/Meis (PM) heterodimers co-operate to regulate enhancer activity in r4. Mutational analysis reveals that these sites are required for activity of the enhancer, suggesting that the r4 enhancer from Hoxa2 functions in vivo as a Hox-response module in combination with the Hox cofactors, Pbx and Prep/Meis. Furthermore, this r4 enhancer is capable of mediating a response to ectopic HOXB1 expression in the hindbrain. These findings reveal that Hoxa2 is a target gene of Hoxb1 and permit us to develop a gene regulatory network for r4, whereby Hoxa2, along with Hoxb1, Hoxb2 and Hoxa1, is integrated into a series of auto- and cross regulatory loops between Hox genes. These data highlight the important role played by direct cross-talk between Hox genes in regulating hindbrain patterning. PMID- 17113576 TI - The sea urchin's siren. AB - This issue of Developmental Biology features articles that constitute a new wave of insights into how a genome interacts with itself (as DNA) and with effectors proteins and probably RNAs, collectively operating as a kind of "cis-trans" dualism. We learned a test for allelism in genetics class that bore that Latin name but now it comes as a new day for biological science-a welcome era in which a phenomenon as complex as development can be envisioned from principles of chemical binding energy and specificity. The buzzword (the term is just-as there is deserved buzz) is that the genome is hard-wired, in the sense that it has been shaped to both encode and react to a regulatory network, of which it is itself a part. I here review some of the milestones of embryology in which the sea urchin was the key player, segueing into the modern era in which this organism launched an entirely new intellectual construct of genome organization and gene expression during development. This essay also contains a number of personal perspectives as well as some views on the overall epistemological fabric of developmental biology. Like all of us, I am excited to see the S. purpuratus genome appear and heartily congratulate, by writing this essay, the trailblazers whose intellectual courage and persistence has brought us to this happy position. PMID- 17113577 TI - Intrinsic differences among spatially distinct neural crest stem cells in terms of migratory properties, fate determination, and ability to colonize the enteric nervous system. AB - We have systematically examined the developmental potential of neural crest stem cells from the enteric nervous system (gut NCSCs) in vivo to evaluate their potential use in cellular therapy for Hirschsprung disease and to assess differences in the properties of postmigratory NCSCs from different regions of the developing peripheral nervous system (PNS). When transplanted into developing chicks, flow-cytometrically purified gut NCSCs and sciatic nerve NCSCs exhibited intrinsic differences in migratory potential and neurogenic capacity throughout the developing PNS. Most strikingly, gut NCSCs migrated into the developing gut and formed enteric neurons, while sciatic nerve NCSCs failed to migrate into the gut or to make enteric neurons, even when transplanted into the gut wall. Enteric potential is therefore not a general property of NCSCs. Gut NCSCs also formed cholinergic neurons in parasympathetic ganglia, but rarely formed noradrenergic sympathetic neurons or sensory neurons. Supporting the potential for autologous transplants in Hirschsprung disease, we observed that Endothelin receptor B (Ednrb)-deficient gut NCSCs engrafted and formed neurons as efficiently in the Ednrb-deficient hindgut as did wild-type NCSCs. These results demonstrate intrinsic differences in the migratory properties and developmental potentials of regionally distinct NCSCs, indicating that it is critical to match the physiological properties of neural stem cells to the goals of proposed cell therapies. PMID- 17113578 TI - New insight into the inhibition of the inflammatory response to experimental delayed-type hypersensitivity reactions in mice by scropolioside A. AB - Scropolioside A, an iridoid isolated from Scrophularia auriculata ssp. pseudoauriculata, showed anti-inflammatory properties against different experimental models of delayed-type hypersensitivity. This iridoid reduced the oedema induced by oxazolone by 79% (72 h) at 0.5 mg/ear while reducing that induced by sheep red blood cells by 47% (18 h), 45% (24 h) and 36% (48 h) at 10 mg/kg. In vivo it reduced both oedema formation and cell infiltration whereas in vitro it reduced the proliferation of activated T-lymphocytes (IC50 of 67.74 microM). Treatment with scropolioside A (100 microM) 18 and 24 h after phytohemagglutinin stimulation increased the number of cells arrested in the subG(0) phase whereas treatment 3 h after stimulation clearly increased the number of cells that passed to the S phase. Scropolioside A also inhibited the production of prostaglandin E2, leukotriene B4, nitric oxide, interleukin-1beta, interleukin-2, interleukin-4, tumour necrosis factor-alpha and interferon-gamma, but had no effect on the production of interleukin-10. Moreover, it modified the expression of both nitric oxide synthase-2 and cyclooxygenase-2, as well as the activation of nuclear factor-kappaB in RAW 264.7 macrophages. PMID- 17113579 TI - Peroxisome proliferator-activated receptor alpha-independent effects of peroxisome proliferators on cysteinyl leukotriene production in mast cells. AB - The effects of peroxisome proliferators, the ligands of a nuclear receptor peroxisome proliferator-activated receptor (PPAR) alpha, on cysteinyl leukotriene production were investigated in rodent mast cells. Peroxisome proliferators Wy 14,643 (30 microM) and fenofibrate (100 microM) significantly inhibited the cysteinyl leukotriene production that was induced by antigen (Ag) treatment after overnight sensitization to Ag specific immunoglobulin E (IgE) in a rat basophilic leukemia (RBL)-2H3 mast cell line. Similar inhibition by these drugs was observed in IgE and Ag-treated mouse bone marrow-derived mast cells, A23187-treated RBL 2H3 and A23187-treated mouse peritoneal macrophages. Wy-14,643 (30 microM) and fenofibrate (100 microM) did not affect the release of radioactivity from RBL-2H3 pre-incubated with [(3)H]-arachidonic acid, which is considered an index of phospholipase A(2) activity. Wy-14,643 (30 microM) and fenofibrate (100 microM) did not directly inhibit 5-lipoxygenase activity. Troglitazone was found to directly inhibit the activity of 5-lipoxygenase. The PPARalpha mRNA level was at less than the limit of detection for the realtime polymerase chain reaction both in RBL-2H3 and bone marrow-derived mast cells. Wy-14,643 (30 microM) and fenofibrate (100 microM) did not induce acyl-CoA oxidase mRNA in RBL-2H3, which was reported to be induced by peroxisome proliferators via PPARalpha in hepatocytes. Wy-14,643 (30 microM) and fenofibrate (100 microM) inhibited the cysteinyl leukotriene production in bone marrow-derived mast cells from PPARalpha null mice. It was concluded that the inhibitory effects of these peroxisome proliferators on cysteinyl leukotriene production are independent of PPARalpha in mast cells. PMID- 17113580 TI - Mitochondrial thioredoxin in regulation of oxidant-induced cell death. AB - Mitochondrial thioredoxin (mtTrx) can be oxidized in response to inducers of oxidative stress; yet the functional consequences of the oxidation have not been determined. This study evaluated the redox status of mtTrx and its association to oxidant-induced apoptosis. Results showed that mtTrx was oxidized after exposure to peroxides and diamide. Overexpression of mtTrx protected against diamide induced oxidation and cytotoxicity. Oxidation of mtTrx was also achieved by knocking down its reductase; and lead to increased susceptibility to cell death. The data indicate that the redox status of mtTrx is a regulatory mechanism underlying the vulnerability of mitochondria to oxidative injury. PMID- 17113581 TI - Descending serotonergic facilitation of spinal ERK activation and pain behavior. AB - Serotonin (5-HT) derived from bulbo-spinal projection is released by nociceptive input into the spinal dorsal horn. Here we report that formalin injection in the paw produced pain behavior (flinching) and phosphorylation of spinal ERK1/2 (P ERK1/2, indicating activation) in rats. Depletion of spinal 5-HT by intrathecal (IT) 5,7-DHT, a serotonergic neurotoxin, profoundly reduced formalin evoked flinching and the increase in P-ERK1/2. Ondansetron (a 5-HT3 receptor antagonist) at IT doses that inhibited flinching also attenuated spinal ERK activation. These findings reveal that primary afferent-evoked activation of spinal ERK requires the input from an excitatory 5-HT descending pathway. PMID- 17113582 TI - Different modulation of TRAIL-induced apoptosis by inhibition of pro-survival pathways in TRAIL-sensitive and TRAIL-resistant colon cancer cells. AB - Epithelial cells can be manipulated to undergo apoptosis depending on the balance between pro-survival and apoptotic signals. We showed that TRAIL-induced apoptosis may be differentially regulated by inhibitors of MEK ERK (U0126) or PI3K/Akt (LY294002) pathway in TRAIL-sensitive (HT-29) and TRAIL-resistant (SW620) human epithelial colon cancer cells. U0126 or LY294002 significantly enhanced TRAIL-induced apoptosis in HT-29 cells, but not in SW620 cells. We report a different regulation of the level of an anti-apoptotic Mcl-1 protein under MEK/ERK or PI3K/Akt pathway inhibition and suggest the mechanisms involved. A special attention was paid to the role of the ERK1/2, Akt, and glycogen synthase kinase 3beta. PMID- 17113583 TI - Pan-genome isolation of low abundance transcripts using SAGE tag. AB - The SAGE (serial analysis of gene expression) method is sensitive at detecting the lower abundance transcripts. More than a third of human SAGE tags identified are novel representing the low abundance unknown transcripts. Using the GLGI method (generation of longer 3' EST from SAGE tag for gene identification), we converted 1009 low-copy, human X chromosome-specific SAGE tags into 10210 3' ESTs. We identified 3418 unique 3' ESTs, 46% of which are novel and originated from the lower abundance transcripts. However, nearly all 3' ESTs were mapped to various regions across the genome but not X chromosome. Detailed analysis indicates that those 3' ESTs were isolated by SAGE tag mis-priming to the non parent transcripts. Replacing SAGE tags with non-transcribed genomic DNA tags resulted in poor amplification, indicating that the sequence similarity between different transcripts contributed to the amplification. Our study shows the prevalence of novel low abundance transcripts that can be isolated efficiently through SAGE tags mis-priming. PMID- 17113584 TI - Rat RFamide-related peptide-3 stimulates GH secretion, inhibits LH secretion, and has variable effects on sex behavior in the adult male rat. AB - A recently described avian neuropeptide, gonadotropin inhibitory hormone (GnIH), has been shown to have seasonal regulatory effects on the hypothalamic-pituitary gonadotropin axis (HPG) in several avian species. In the bird, GnIH expression is increased during the photorefractory period and has inhibitory effects on the HPG. A recently described mammalian neuropeptide, RF-amide-related peptide-3 (RFRP-3), may be genetically related and functionally similar to this avian neuropeptide. The purposes of this study were to first see if rat RFRP-3 is expressed in the male rat brain and second to determine if ICV injections of RFRP 3 will have effects on feeding and sex behaviors, as well as hormone release from the anterior pituitary. Results confirm other studies in that immunoreactive cell bodies and fibers are observable in areas of the male rat brain known to control the HPG and feeding and sex behaviors. RFRP-3 fibers are also observed in close proximity to GnRH immunoreactive cell bodies. Behavioral tests indicate that high but not low ICV RFRP-3 (500 vs. 100 ng, respectively) significantly (p<0.05) suppressed all facets of male sex behavior while not having any observable effects on their ability to ambulate. Sex behavior was later exhibited when those same male rats received the ICV vehicle. While suppressing sex behavior, ICV RFRP 3 significantly (p<0.05) increased food intake compared to controls. ICV RFRP-3 also significantly reduced plasma levels of luteinizing hormone but increased growth hormone regardless of the time of day; however, at no time did RFRP-3 alter plasma levels of FSH, thyroid hormone, or cortisol. These results indicate that although RFRP-3 has similar effects on LH as observed with GnIH in avian species, in the rat RFRP-3 has additional roles in regulating feeding and growth. PMID- 17113585 TI - Use of high-resolution MRI for investigation of fluid flow and global permeability in a material with interconnected porosity. AB - We present a multi-scale experimental approach designed to improve the investigation of both localized and global fluid flow in biomaterials with randomly interconnected porosity. Coralline hydroxyapatite (ProOsteon 500 from Interpore-Cross), having a relatively well-defined porosity, was used as an in vitro model of typical bone architecture. Axial fluid velocity profiles within the pores of a cylindrical hydroxyapatite sample were characterized using high resolution MRI in conjunction with the measurement of global flow and associated permeability based on the Darcy-type relationship. Assuming Newtonian fluid behaviour, image analysis permitted computation of local porosity, intra-pore fluid shear, and visualization of flow heterogeneity within the sample. These results may benefit applications in biomaterials for the evaluation of factors influencing bony incorporation in porous scaffolds and on porous implant and bone surfaces. Normal and diseased biological tissues are also clinical relevant applications. PMID- 17113586 TI - Determination of DNA entrapment into liposomes using short monolithic columns. AB - A high-performance liquid chromatography (HPLC) method for the determination of DNA entrapment efficiency in liposomes has been developed. Plasmid DNA was encapsulated into positively charged liposomes. Non-entrapped DNA was separated by ultracentrifugation from liposomes and supernatant was chromatographed on Convective Interaction Media (CIM) DEAE disk. The elution of DNA was monitored by the absorbance at 260 nm and the quantity of DNA in the tested sample was calculated from the integrated peak areas using the appropriate standard curve. This method is fast, simple, precise and does not require any kind of DNA labelling in contrast with mostly used methods for determination of DNA entrapment efficiency. PMID- 17113587 TI - Preparation of convection interaction media isobutyl disc monolithic column and its application to purification of secondary alcohol dehydrogenase and alcohol oxidase. AB - A convection interaction media (trade name CIM, BIA Separation, Ljubljana, Slovenia) isobutyl monolithic disc was prepared by incubating a CIM epoxy monolithic disc with isobutylamine, and it was then applied to the purification of secondary alcohol dehydrogenase (S-ADH) and primary alcohol oxidase (P-AOD). Both enzymes were adsorbed on this column and eluted with high purity. Thus, S ADH was purified to an electrophoretically homogeneous state by four column chromatographies using CIM DEAE-8 and CIM C4-8 tube monolithic columns, blue Sepharose column and CIM isobutyl disc monolithic column. P-AOD was also purified to an electrophoretically homogeneous state by three column chromatographies of CIM DEAE-8 tube, CIM C4-8 tube and CIM isobutyl disc columns. PMID- 17113588 TI - Nonaqueous capillary electrophoretic behavior of 2-aryl propionic acids in the presence of an achiral ionic liquid. A chemometric approach. AB - Ionic liquids (ILs) appear really attractive as electrolyte additives in nonaqueous capillary electrophoresis (NACE). These salts may offer new possibilities of interactions to modulate analyte effective mobilities. The presence of 1-n-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide (BMIM NTf2) in acetonitrile/alcohol background electrolytes (BGEs) was investigated in this work. The aim of this study was to elucidate the influence of the IL concentration on the electrophoretic behavior of four arylpropionic acids and to identify the interactions between the analytes and the IL cation. The influence on mobility of the IL concentration, the nature and the proportion of the organic solvents, and the concentration of the ionic components of the BGE was first studied by a univariate approach. A four-factor D-optimal experimental design was then applied to provide a deeper insight into analyte interaction with IL cation present both free in BGE and adsorbed onto the capillary wall. PMID- 17113589 TI - Direct determination of chlorophenols in environmental water samples by hollow fiber supported ionic liquid membrane extraction coupled with high-performance liquid chromatography. AB - The 1-octyl-3-methylimidazolium hexafluorophosphate ([C8MIM][PF6]) ionic liquid was immobilized in the pores of a polypropylene hollow fiber for hollow fiber protected liquid-phase microextraction. Analytes including 4-chlorophenol (4-CP), 3-chorophenol (3-CP), 2,4-dichlorophenol (2,4-DCP) and 2,4,6-trichlorophenol (2,4,6-TCP) were extracted into this ionic liquid membrane, and back extracted into 10microL sodium hydroxide acceptor solution in the lumen of the hollow fiber. Then, the acceptor solution was withdrawn into the high-performance liquid chromatography (HPLC) microsyringe connected to the hollow fiber, and directly injected into the HPLC system for analysis. Some parameters that might affect the extraction efficiency were optimized, and low detection limits (0.5microgL(-1) for 4-CP, 3-CP, DCP and 1.0microgL(-1) for TCP) were obtained. Good repeatability was achieved because of the stability of the hollow fiber-supported ionic liquid membrane. The proposed procedure was applied for direct determination of the four chlorophenols in some real water samples including groundwater, river water, wastewater and tap water. All of the four chlorophenols in these water samples were under the limits of determination, and the recoveries were in the range of 70.0-95.7% at 5microgL(-1) spiked level. PMID- 17113590 TI - Characterisation of UV-cured acrylate networks by means of hydrolysis followed by aqueous size-exclusion combined with reversed-phase chromatography. AB - UV-cured networks prepared from mixtures of di-functional (polyethylene-glycol di acrylate) and mono-functional (2-ethylhexyl acrylate) acrylates were analysed after hydrolysis, by aqueous size-exclusion chromatography coupled to on-line reversed-phase liquid-chromatography. The mean network density and the fraction of dangling chain ends of these networks were varied by changing the concentration of mono-functional acrylate. The amount and the molar-mass distribution of the polyethylene-glycol chains between cross-links (M(XL)) and polyacrylic acid (PAA) backbone chains (the so-called kinetic chain length (kcl)) in the different acrylate networks were determined quantitatively. The molar-mass distribution of kcl revealed an almost linear dependence on the concentration of mono-functional acrylate. Analysis of the starting materials showed a significant concentration of mono-functional polyethylene-glycol acrylate. In combination with the analysis of the extractables of the UV-cured networks (polymers not attached to the network, impurities that originate from the photo-initiator and unreacted monomers), more insight in the total network structure was obtained. It was shown that the UV-cured networks contain only small fractions of residual compounds. With these results, the chemical network structure for the different UV-cured acrylate polymers was expressed in network parameters such as the number of PAA units which are cross-linked, the degree of cross-linking, and the network density, which is the molar concentration of effective network chains between cross-links per volume of the polymers. The mean molar mass of chains between chemical network junctions (M(C)) was calculated and compared with results obtained from solid-state NMR and DMA. The mean molar mass of chains between network junctions as determined by these methods was similar. PMID- 17113591 TI - Time-weighted average water sampling with a diffusion-based solid-phase microextraction device. AB - A new diffusion-based solid-phase microextraction (SPME) time-weighted average (TWA) field water sampling device was developed and investigated by field trial. The sampler is constructed with copper tube and caps and a commercial SPME fiber assembly. The device possesses all advantages of SPME; it is solvent-free, reusable, combines sampling, isolation and enrichment into one step, and the fiber can be directly injected into a gas chromatograph for analysis with a commercial SPME fiber holder, without further treatment. Field trials in Laurel Creek (Waterloo, Ont., Canada) and Hamilton Harbour (Hamilton, Ont., Canada) illustrated that the device is durable, easy to deploy, and the mass uptake of the device is independent of the face velocity. The device provides good precision [relative standard deviations (RSDs) are less than 20%] and the data obtained with this device are quite comparable to those obtained with the spot sampling method, which demonstrates that the newly developed SPME water sampling device is suitable for long-term monitoring of organic pollutants in water. PMID- 17113592 TI - Improved single-drop microextraction for high sensitive analysis. AB - This paper described a simple approach to prepare a small bell-mouthed extraction device for single-drop microextraction (SDME). Analytical sensitivity was improved by increasing the suspended acceptor volume. Because of the increased contact area and the rough inner surface of the extraction device, the stability of drop was markedly increased. The merits of the proposed method were demonstrated by using 1-octanol as extractant and with cyanazine, simazine and atrazine as model compounds. The related parameters and the effect of humic acid were systematically investigated. Under the optimized extraction conditions, the linear range, detection limit (S/N=3) and precision (RSD, n=6) were 0.2-50, 0.06microgL-1, 5.7% for cyanazine, 0.1-25, 0.03microgL-1, 6.7% for simazine, and 0.15-37.5, 0.04microgL-1, 5.0% for atrazine, respectively. The established method was applied to determine the target compounds in four real water samples, and the satisfactory spiked recoveries at two concentration levels were obtained. Moreover, the comparison of the proposed SDME with the traditional SDME was performed. These results indicated that the proposed improvement made SDME be a competitive analytical tool and an alternative of the traditional methods for the analysis of organic pollutants at trace level. PMID- 17113593 TI - Salt effect on the interactions between gemini surfactant and oppositely charged polyelectrolyte in aqueous solution. AB - The effect of alkali halides (NaBr, NaCl, KCl) on the interactions between the cationic gemini surfactant hexylene-1,6-bis(dodecyldimethylammonium bromide) (12 6-12) and the anionic polyelectrolyte sodium polyacrylate (NaPAA) in aqueous solution has been investigated by fluorescence emission spectroscopy, UV transmittance, zeta potential, and transmission electron microscopy (TEM). With increased addition of NaBr, a counterbalancing salt effect on the critical aggregation concentration (CAC) is observed. At low concentrations, NaBr facilitates the formation of micelle-like structures between surfactant and polyelectrolyte and results in a smaller CAC. At high concentrations, NaBr screens the electrostatic attraction between surfactant and polyelectrolyte and leads to a larger CAC. Upon the formation of micelle-like structures at high surfactant concentrations, the addition of NaBr is favorable for larger aggregates. The microstructure detected by TEM show that a global structure is generally formed in the presence of NaBr. The interactions also depend on ion species. Compared to NaBr, the addition of NaCl or KCl yields a smaller CAC. PMID- 17113594 TI - Effect of dispersion pH on the formation and stability of Pickering emulsions stabilized by layered double hydroxides particles. AB - Using positively charged plate-like layered double hydroxides (LDHs) particles as emulsifier, liquid paraffin-in-water emulsions stabilized solely by such particles are successfully prepared. The effects of the pH of LDHs aqueous dispersions on the formation and stability of the emulsions are investigated here. The properties of the LDHs dispersions at different pHs are described, including particle zeta potential, particle aggregation, particle contact angle, flow behavior of the dispersions and particle adsorption at a planar oil/water interface. The zeta potential decreases with increasing pH, leading to the aggregation of LDHs particles into large flocs. The structural strength of LDHs dispersions is enhanced by increasing pH and particle concentration. The three phase contact angle of LDHs also increases with increasing pH, but the variation is very small. Visual observation and SEM images of the interfacial particle layers show that the adsorption behavior of LDHs particles at the planar oil/water interface is controlled by dispersion pH. We consider that the particle particle (at the interface) and particle-interface electrostatic interactions are well controlled by adjusting the dispersion pH, leading to pH-tailored colloid adsorption. The formation of an adsorbed particle layer around the oil drops is crucial for the formation and stability of the emulsions. Emulsion stability improves with increasing pH and particle concentration because more particles are available to be adsorbed at the oil/water interface. The structural strength of LDHs dispersions and the gel-like structure of emulsions also influence the stability of the emulsions, but they are not necessary for the formation of emulsions. The emulsions cannot be demulsified by adjusting emulsion pH due to the irreversible adsorption of LDHs particles at the oil/water interface. TEM images of the emulsion drops show that a thick particle layer forms around the oil drops, confirming that Pickering emulsions are stabilized by the adsorbed particle layers. The thick adsorbed particle layer may be composed of a stable inner particle layer which is in direct contact with the oil phase and a relatively unstable outer particle layer surrounding the inner layer. PMID- 17113595 TI - Gluttony and sex in female ixodid ticks: how do they compare to other blood sucking arthropods? AB - The central issue dealt with here is the role of copulation in the control of feeding behaviour in ticks and some haematophagous insects. Female ticks of the family Ixodidae normally engorge to approximately 100 x their unfed body weight, and then drop from the host, produce and lay eggs, and die. Virgins, on the other hand, normally do not exceed 5-40% (depending on species) of the normal engorged body weight. But instead of detaching voluntarily at that point most virgins remain fixed to the host for extended periods, waiting for males to find them so they can complete engorgement. Virgin haematophagous insects, and virgin ticks of the family Argasidae display little, if any, reduction in blood meal size compared to mated females, at least not during the first ovarian cycle. During subsequent ovarian cycles, meal size in some virgin insects may be somewhat reduced depending on how many eggs are retained in the reproductive tract, but the reduction is not nearly to the same extent as that observed for virgin ixodid females. The stimulatory effect of copulation on engorgement in the latter is caused by a pair of proteins (voraxin alpha and beta) produced in the testis and transferred to the female with the spermatophore. Here, I propose why it might be adaptive for an ixodid female to remain small until mated. The hypothesis is suggested from the facts that ixodid ticks remain attached to the host for days (rather than minutes), and that virgin ticks, above a certain critical weight, lose all opportunity for producing viable offspring should they be groomed off the host prematurely, or should the host die while ticks are still attached. PMID- 17113596 TI - Domain interdependence in the biosynthetic assembly of CFTR. AB - The dimerization of their two nucleotide binding domains (NBDs) in a so-called "nucleotide-sandwich" is the hallmark of ATP cassette binding (ABC) proteins and the basis of their catalytic activities. The major disease-causing mutation in the cystic fibrosis transmembrane conductance regulator (CFTR or ABCC7), deletion of Phe508 in NBD1, does not grossly alter the structure of that domain but prevents conformational maturation of the whole CFTR protein, possibly by disrupting the native interaction between NBD1 and NBD2. However, the role of inter-domain interactions in CFTR folding has been brought into question by a recent report that all CFTR domains fold independently. Here we show that in addition to domain folding, correct inter-domain assembly is essential to form a stable unit that satisfies endoplasmic reticulum (ER) quality control. N-terminal domains depend on their more C-terminal neighbors, most essentially the second membrane-spanning domain (MSD2) but significantly, not NBD2. Wild-type C-terminal truncation constructs, completely devoid of NBD2 are transported out of the ER and to the cell surface where they form characteristic CFTR chloride channels with low open probability. The DeltaNBD2 wild-type protein matures and has similar stability as its full-length counterpart. Therefore, the catalytically crucial inter-NBD associations are not required to satisfy ER quality control mechanisms. The DeltaF508 mutation arrests the maturation of DeltaNBD2 just as it does full-length CFTR, indicating that DeltaF508 perturbs other portions of the molecule in addition to NBD2. We find that the mutation prevents formation of a compact MSD1, reflected in its susceptibility to protease digestion. This perturbation of MSD1 may in turn prevent its normal integration with MSD2. The dispensability of NBD2 in the folding of more N-terminal domains stands in contrast to the known hypersensitivity to proteolysis of NBD2 in the DeltaF508 protein. PMID- 17113597 TI - The mechanosensitive channel protein MscL is targeted by the SRP to the novel YidC membrane insertion pathway of Escherichia coli. AB - The mechanosensitive channel MscL in the inner membrane of Escherichia coli is a homopentameric complex involved in homeostasis when cells are exposed to hypo osmotic conditions. The E. coli MscL protein is synthesized as a polypeptide of 136 amino acid residues and uses the bacterial signal recognition particle (SRP) for membrane targeting. The protein is inserted into the membrane independently of the Sec translocon. Mutants affected in the Sec-components are competent for MscL assembly. Translocation of the periplasmic domain was detected using a membrane-impermeant, sulfhydryl-specific gel-shift reagent. The modification of a single cysteine residue at position 68 indicated its translocation across the inner membrane. From these in vivo experiments, it is concluded that the electrical chemical membrane potential is not necessary for membrane insertion of MscL. However, depletion of the membrane insertase YidC inhibits translocation of the protein across the membrane. We show here that YidC is essential for efficient membrane insertion of the MscL protein. YidC is a component of a recently identified membrane insertion pathway that is evolutionarily conserved in bacteria, mitochondria and chloroplasts. PMID- 17113598 TI - Cerebrovascular reactivity in depressed patients without vascular risk factors. AB - INTRODUCTION: Cerebrovascular reactivity (CVR) seems to be gaining importance as a prognostic factor for stroke risk. CVR reflects the compensatory dilatory capacity of cerebral arterioles to a dilatory stimulus; this mechanism plays an important role in maintaining a constant cerebral blood flow. Evaluating factors that influence CVR will help prevention or early detection of cerebrovascular disease (CVD). In this study we aimed to measure the CVR in vascular-risk free depressed individuals so as to evaluate the effect depression has on CVR and hence its role as a stroke risk factor. METHODS: Using acetazolamid (ACZ) stimulation, CVR was assessed with a transcranial Doppler ultrasound in 25 non smoking depressed patients (average age: 48.48 +/- 14.40) and in 25 healthy non smoking controls (average age: 46.76 +/- 13.69) by calculating the difference between the maximal mean blood flow velocity at baseline and the maximal mean blood flow velocity after ACZ stimulation. RESULTS: Basal Cerebral Blood flow in Patients was 50.6 cm/s (SD: 11.6) versus controls 52.80 cm/s (SD: 12.70) whereas after stimulation maximal blood flow velocity was 72.64 cm/s (SD: 15.75) in patients versus 80.20 cm/s (SD: 18.43) in controls. In an analysis of covariance we found that cerebrovascular reactivity was significantly reduced in the vascular-risk free depressed sample. Age had a significant influence whereas gender did not. DISCUSSION: Major Depression appears to decrease cerebrovascular reactivity supporting the idea of increased risk for stroke in depressed patients. The mechanisms leading to this phenomenon and its subtle subgroup differences should be further investigated. PMID- 17113599 TI - Associations between MDR1 gene polymorphisms and schizophrenia and therapeutic response to olanzapine in female schizophrenic patients. AB - Multidrug resistant protein (MDR1) gene, which codes for P-glycoprotein and functions as an efflux transporter in different cells, is widely localized in normal tissues including the gastrointestinal tract, blood cells, biliary tract, kidney and brain and plays a major role in absorption, distribution and elimination of various xenobiotics. Therefore, MDR1 gene variants were proposed as potential susceptibility factors for diseases and as determinants of treatment response to various drugs. We investigated the relationships between exon 21 G2677T and exon 26 C3435T genetic variants of MDR1 gene with susceptibility and treatment response in female schizophrenic patients. The study was conducted in two steps. We first compared allele, genotype and haplotype distributions between 117 female schizophrenic patients and 123 control female subjects. Afterwards, we studied treatment response to olanzapine, in 87 out of 117 previously unmedicated female patients. Overall, we found lower representation of G2677/C3435 haplotype in schizophrenic female patients compared to controls. Test result for linkage disequilibrium between loci was found to be significant. Furthermore, we found significant associations between MDR1 exon 21 G2677T genotypes and treatment response measured with positive PANSS percentage changes, with T allele and TT genotype being associated with significantly better treatment response. A borderline, non-significant statistical association was found between MDR1 exon 26 C3435T genotypes and treatment response, with TT genotype being associated with better treatment response. Our data support functional importance of the MDR1 mutations for the susceptibility and treatment response in female schizophrenic patients. PMID- 17113600 TI - Epidemiology of depression and its treatment in the general population. AB - This study examines the correlates of a major depressive disorder and its treatment in the general population. The sample was composed of 6694 individuals aged between 18 and 96 years, representative of the general population of the states of California and New York (48 million inhabitants aged 18 years or older). They were interviewed by telephone using the Sleep-EVAL system. The interviews included various sleep and health topics and the assessment of DSM-IV sleep and psychiatric disorders. The 1-month prevalence of a major depressive disorder was 5.2% in the sample, and was higher in women, middle-aged and non Hispanic white individuals. Obesity (BMI > or =30kg/m(2)), poor health status and smoking were also strongly correlated with a major depressive disorder. A total of 57.7% of depressed subjects were receiving some forms of treatment for depression: 28.3% were taking antidepressants (alone or in combination with psychiatric health care) and 29.4% received psychiatric health care (without antidepressant medication). Severity of depression, ethnicity and weight (overweight or obese) were strongly associated with the presence of treatment. A major depressive disorder is frequent in the general population. Although its identification and treatment have improved over the years, some segments of the population, namely elderly and non-white individuals are less likely to receive appropriate care. PMID- 17113601 TI - Consumer product-related eye injury in the United States, 1998-2002. AB - PROBLEM: Eye injury is currently a leading cause of visual impairment and monocular blindness in the United States. Information regarding consumer products associated with eye injuries can have important implications for the prevention of these injuries. METHODS: The National Electronic Injury Surveillance System (NEISS) was used to describe the types of consumer products associated with emergency department treated eye injuries in the United States from 1998 through 2002. RESULTS: The leading product type associated with eye injuries was welding equipment followed by household cleaners, basketball equipment, workshop equipment, and adhesives. Eye injuries attributed to hardware, tools, construction, sports, toys, and lawn equipment were more common among males. In females, eye injuries attributable to chemicals, housewares, storage and organization, and bed and bath items were more common. Differences were also apparent across the age spectrum. CONCLUSION: This study identified specific products and categories of products frequently associated with eye injury and prevention initiatives should focus on these items. IMPACT ON INDUSTRY: This study has identified consumer products associated with eye injuries requiring medical treatment in the United States. Manufacturers of these products could be encouraged to add or strengthen safety messages regarding the potential for eye injury. PMID- 17113602 TI - Lipoprotein (a) and risk of ischemic stroke in young adults. AB - Lipoprotein (a) [Lp(a)] is a LDL-particle linked to apoprotein (a) [apo(a)]. High Lp(a) plasma level is a risk factor for coronary heart disease and, in older men, for ischemic stroke. The role of Lp(a) as a risk factor for ischemic stroke in young adults is uncertain. METHODS: Lp(a) concentration was prospectively measured in 100 consecutive patients with acute ischemic stroke (58 men and 42 women) aged 18-55 years, and in 100 controls matched for age and gender. RESULTS: The distribution of Lp(a) concentration was skewed toward the highest and median tertiles in male patients. In multivariate logistic regression analyses adjusting on classical risk factors for ischemic stroke and lipid variables, Lp(a) concentration in the highest and medium tertiles compared with the lowest tertile was significantly associated with ischemic stroke in men (OR 3.55, 95% CI 1.33 9.48, p = 0.012), but was not in women (OR 0.42, 95% CI 0.14-1.26, p = 0.12). Although large vessel atherosclerosis was more common in men than in women, there were no differences in Lp(a) concentration according to the cause of ischemic stroke. CONCLUSION: Among subjects aged 18-55 years, a slightly elevated Lp(a) concentration was strongly and independently associated with ischemic stroke in men, but not in women. Further studies are required to elucidate the mechanisms underlying this gender-specific association. PMID- 17113603 TI - No association between MTHFR A1298C and MTRR A66G polymorphisms, and MS in an Australian cohort. AB - Multiple sclerosis (MS) is a complex neurological disease that affects the central nervous system (CNS) resulting in debilitating neuropathology. Pathogenesis is primarily defined by CNS inflammation and demyelination of nerve axons. Methionine synthase reductase (MTRR) is an enzyme that catalyzes the remethylation of homocysteine (Hcy) to methionine via cobalamin and folate dependant reactions. Cobalamin acts as an intermediate methyl carrier between methylenetetrahydrofolate reductase (MTHFR) and Hcy. MTRR plays a critical role in maintaining cobalamin in an active form and is consequently an important determinant of total plasma Hcy (pHcy) concentrations. Elevated intracellular pHcy levels have been suggested to play a role in CNS dysfunction, neurodegenerative, and cerebrovascular diseases. Our investigation entailed the genotyping of a cohort of 140 cases and matched controls for MTRR and MTHFR, by restriction length polymorphism (RFLP) techniques. Two polymorphisms: MTRR A66G and MTHFR A1298C were investigated in an Australian age and gender matched case control study. No significant allelic frequency difference was observed between cases and controls at the alpha = 0.05 level (MTRR chi2 = 0.005, P = 0.95, MTHFR chi2 = 1.15, P = 0.28). Our preliminary findings suggest no association between the MTRR A66G and MTHFR A1298C polymorphisms and MS. PMID- 17113604 TI - Human heart mitochondria do not produce physiologically relevant quantities of nitric oxide. AB - Previous studies raised the possibility that nitric oxide synthase is present in heart mitochondria (mtNOS) and the existence of such an enzyme became generally accepted. However, original experimental evidence is rather scarce and positive identification of the enzyme is lacking. We aimed to detect an NOS protein in human and mouse heart mitochondria and to measure the level of NO released from the organelles. Western blotting with 7 different anti-NOS antibodies failed to detect a NOS-like protein in mitochondria. Immunoprecipitation or substrate affinity purification of the samples concentrated NOS in control preparations but not in mitochondria. Release of NO from live respiring human mitochondria was below 2 ppb after 45 min of incubation. In a bioassay system, mitochondrial suspension failed to cause vasodilation of human mammary artery segments. These results indicate that mitochondria do not produce physiologically relevant quantities of NO in the heart and are unlikely to have any physiological importance as NO donors, nor do they contain a recognizable mtNOS enzyme. PMID- 17113605 TI - Physiological and therapeutic factors affecting cholesterol metabolism: does a reciprocal relationship between cholesterol absorption and synthesis really exist? AB - Cholesterol absorption and synthesis contribute to maintaining cholesterol homeostasis. Several physiological and therapeutic factors affect cholesterol homeostasis, including: genetics, circadian rhythm, body weight, plant sterols, ezetimibe, and statin therapy. The present objective is to determine the main vector, i.e. cholesterol absorption or synthesis, affected by each of these factors, and to examine whether an alteration in one vector is linked to a reciprocal change in the other. Current techniques used to assess cholesterol absorption and synthesis are also reviewed. Review of physiological factors affecting cholesterol metabolism suggest a reciprocal relationship between these two vectors. Carriers of the E2 isoform of apolipoprotein E and ATP binding cassette (ABC) G8 19H (exon 1 mutation) show a decrease in cholesterol absorption accompanied by a corresponding increase in synthesis. Circadian rhythm affects cholesterol synthesis, however, its effect on absorption has yet to be established. Obese subjects show an increase in cholesterol synthesis with a subsequent decrease in cholesterol absorption. Weight loss down regulates cholesterol synthesis, but has little or no effect on absorption. In the case of therapeutic factors, plant sterols and stanols inhibit cholesterol absorption, which results in a compensatory increase in synthesis. Ezetimibe also decreases intestinal absorption, while reciprocally increasing synthesis. Statin therapy down regulates synthesis, which is accompanied by a rise in absorption. These findings suggest that a change in one vector, fairly consistently, results in a compensatory and opposing change in the other. An understanding of this reciprocal relationship between cholesterol absorption and synthesis may allow for the development of more effective interventions for dyslipidemic disorders. PMID- 17113606 TI - Urban seagrass: status of Posidonia oceanica facing the Genoa city waterfront (Italy) and implications for management. AB - A system of five adjacent Posidonia oceanica meadows facing the waterfront of Genoa city (Ligurian Sea, NW Mediterranean) was investigated over different spatial scales (meters-kilometers) using three environmental indices: conservation index (CI), substitution index (SI) and phase-shift index (PSI). CI revealed differences mostly at large spatial scale, distinguishing the poor condition of the meadows closest to Genoa centre and harbour from the comparatively healthy condition of the farthest meadows. SI showed differences mostly at small spatial scale (i.e., within meadows), suggesting the influence of local factors in the re-colonisation of regressed meadows by the seagrass Cymodocea nodosa and/or the invasive alga Caulerpa racemosa. Mapping of PSI showed that the meadows closest to Genoa centre and harbour have undergone a nearly total phase shift and have no real potential for recovery: attempts to re establish P. oceanica there might be a waste of time and money. On the contrary, the meadows farthest from Genoa centre and harbour showed a comparatively low level of phase shift and could still fully recover given specific management actions. PMID- 17113607 TI - Spatial distribution and species composition of PAHs in surface sediments from the Bohai Sea. PMID- 17113608 TI - A comparison of acute and chronic toxicity tests used to examine the temporal stability of a gradient in copper tolerance of Hediste diversicolor from the Fal estuary, Cornwall, UK. AB - The aim of this study was to use two different toxicity tests to verify the existence of a gradient in tolerance along Rostronguet Creek. Hediste diversicolor was collected from five populations in the Fal estuary previously shown to vary in copper tolerance. Exposure to 4 mgL(-1) of copper in an acute assay demonstrated that Mylor Creek worms were sensitive (LT(50) 86 h) and the tolerance of Rostronguet Creek worms increased moving upstream from the mouth of the creek (LT(50)s 100-258 h). There was no significant difference in tolerance between Mylor worms and worms from the mouth of Rostronguet Creek. This is in agreement with a previous study [Grant, A., Hateley, J.G., Jones, N.V., 1989. Mapping the ecological impact of heavy metals on the estuarine polychaete Nereis diversicolor using inherited metal tolerance. Marine Pollution Bulletin 20, 235 238] and demonstrates temporal stability of the gradient. Copper tolerance was also measured using a chronic toxicity test run for 90 d using step-wise increases in challenge concentration. A significant difference in tolerance was shown between populations from Mylor Creek and those at the mouth of Rostronguet Creek, which has not been reported previously. Experimental protocol was therefore an important factor in detecting population variation in tolerance. PMID- 17113609 TI - Chemical contamination of the Hudson-Raritan Estuary as a result of the attack on the World Trade Center: analysis of polycyclic aromatic hydrocarbons and polychlorinated biphenyls in mussels and sediment. AB - The September 11, 2001 attack on the World Trade Center (WTC) resulted in a massive plume of dust and smoke that blanketed lower Manhattan and part of the Hudson-Raritan Estuary (HRE). The NOAA National Status and Trends Mussel Watch Program has long-term monitoring sites in the area and thus had an opportunity to assess the effect of the WTC attack on PAH and PCB contamination of the surrounding estuary. Seven additional sites were added in the Upper HRE to attain higher sampling resolution for comparison with regularly sampled Mussel Watch Project HRE sites. Elevated background levels of PCBs and PAHs in mussel tissue and sediments were high enough before the WTC attack that concentrations were not measurably changed by WTC derived contaminant input. PMID- 17113610 TI - Emotional & electroencephalographic responses during affective picture viewing after exercise. AB - We examined the effects of 30 min of cycling exercise at a moderate intensity of 50% peak oxygen uptake, compared to 30 min of rest, on changes in emotional responses to pictorial foreground stimuli that reliably elicit unpleasant, neutral, and pleasant affect. Emotional responses were measured by self-reports of valence (unpleasant to pleasant) and arousal (low to high) and by hemispheric asymmetry (R-L) of frontal and parietal brain electroencephalographic (EEG) activity in 13 females and 21 males (24+/-3 y). Compared to after rest, self reports of arousal in response to unpleasant slides were diminished after exercise, but self-reports of valence and frontal asymmetry of alpha frequencies were generally unchanged. Even so, there were differential responses in asymmetry in the beta frequencies in the frontal region and for alpha and beta frequencies in the parietal region, indicative of decreased activity in the left frontal and right parietal regions after exercise compared to after rest. We conclude that moderately intense cycling exercise generally does not alter emotional responding to pleasant and neutral pictures, but may reduce emotional arousal during exposure to unpleasant stimuli. PMID- 17113611 TI - Preparative enzymatic solid phase synthesis of cis(+)-12-oxo-phytodienoic acid - physical interaction of AOS and AOC is not necessary. AB - The pathway of jasmonic acid (JA) biosynthesis was established in the 1980s by Vick and Zimmerman but, until now, the preparative biosynthesis of the jasmonic acid precursors 12-oxo-phytodienoic acid (OPDA) and 3-oxo-2-[2'-pentenyl] cyclopentan-1-octanoic acid (OPC-8:0) in their endogenous and biologically relevant cis(+)-configuration was only possible in small amounts and had to put up with high costs. This was mainly due to the lack of high amounts of pure and enzymatically active allene oxide cyclase (AOC), which is a key enzyme in the biosynthesis of jasmonates in that it releases, in a coupled reaction with allene oxide synthase (AOS), the first cyclic and biological active metabolite - OPDA. We describe here the expression and purification of AOS and AOC and their subsequent coupling to solid matrices to produce an enantioselective, reusable bioreactor for octadecanoid production. With the method described here it is possible to produce optically pure enantiomers of octadecanoids in high amounts in a cost- and time-efficient manner. Furthermore, it could be demonstrated that a physical interaction of AOS and AOC, hitherto postulated to be required for substrate channeling from AOS to AOC, is not necessary for the in vitro cyclization of the unstable epoxide generated by the AOS reaction. PMID- 17113612 TI - Poppy alkaloid profiling by electrospray tandem mass spectrometry and electrospray FT-ICR mass spectrometry after [ring-13C6]-tyramine feeding. AB - Papaver alkaloids play a major role in medicine and pharmacy. In this study, [ring-(13)C(6)]-tyramine as a biogenetic precursor of these alkaloids was fed to Papaver somniferum seedlings. The alkaloid pattern was elucidated both by direct infusion high-resolution ESI-FT-ICR mass spectrometry and liquid chromatography/electrospray tandem mass spectrometry. Thus, based on this procedure, the structure of about 20 alkaloids displaying an incorporation of the labeled tyramine could be elucidated. These alkaloids belong to different classes, e.g. morphinan, benzylisoquinoline, protoberberine, benzo[c]phenanthridine, phthalide isoquinoline and protopine. The valuable information gained from the alkaloid profile demonstrates that the combination of these two spectrometric methods represents a powerful tool for evaluating biochemical pathways and facilitates the study of the flux of distant precursors into these natural products. PMID- 17113613 TI - Changes in the nasal carriage of drug-resistant Streptococcus pneumoniae in urban and rural Vietnamese schoolchildren. AB - Studying the antimicrobial drug resistance of nasopharyngeal or nasal carriage isolates of Streptococcus pneumoniae in children is likely to have predictive potential for invasive isolates. Streptococcus pneumoniae nasal carriage was studied in 1422 Vietnamese children. Forty-six percent of 536 isolates showed reduced susceptibility to penicillin and 7% showed intermediate susceptibility to ceftriaxone; and 50% of 518 isolates showed resistance to erythromycin. All isolates were sensitive to levofloxacin and gatifloxacin. Urban and suburban children were significantly more likely to carry drug-resistant isolates than rural children. Rates of non-susceptibility to penicillin and erythromycin increased significantly in the rural province Khanh Hoa in 2003/2004 compared with rates obtained in 1997. An emerging clone of penicillin non-susceptible S. pneumoniae of serogroup 15 was identified, which was widely distributed in addition to the pandemic clones Spain(23F)-1 and Taiwan(19F)-14. Although resistance to fluoroquinolones was not observed, 6 (18%) of 34 isolates had a Lys137Asn mutation in the parC gene. This study shows that drug resistance is increasing in carriage isolates of S. pneumoniae in rural areas in Vietnam owing to spread of pandemic and emerging resistant clones. PMID- 17113614 TI - Application of metabonomics on an experimental model of fibrosis and cirrhosis induced by thioacetamide in rats. AB - Metabonomics has already been used to discriminate different pathological states in biological fields. The metabolic profiles of chronic experimental fibrosis and cirrhosis induction in rats were investigated using (1)H NMR spectroscopy of liver extracts and serum combined with pattern recognition techniques. Rats were continuously administered with thioacetamide (TAA) in the drinking water (300 mg TAA/L), and sacrificed on 1st, 2nd, and 3rd month of treatment. (1)H NMR spectra of aqueous and lipid liver extracts, together with serum were subjected to Principal Component Analysis (PCA). Liver portions were also subjected to histopathological examination and biochemical determination of malondialdehyde (MDA). Liver fibrosis and cirrhosis were progressively induced in TAA-treated rats, verified by the histopathological examination and the alterations of MDA levels. TAA administration revealed a number of changes in the (1)H NMR spectra compared to control samples. The performance of PCA in liver extracts and serum, discriminated the control samples from the fibrotic and cirrhotic ones. Metabolic alterations revealed in NMR spectra during experimental liver fibrosis and cirrhosis induction, characterize the stage of fibrosis and could be illustrated by subsequent PCA of the spectra. Additionally, the PCA plots of the serum samples presented marked clustering during fibrosis progression and could be extended in clinical diagnosis for the management of cirrhotic patients. PMID- 17113615 TI - The interaction of spider gating modifier peptides with voltage-gated potassium channels. AB - Gating modifier peptides bind to ion channels and alter the gating process of these molecules. One of the most extensively studied peptides, Hanatoxin (HaTx), isolated from a Chilean tarantula, has been used to characterize the blocking properties of the voltage-gated potassium channel Kv2.1. These studies have provided some insight into the gating mechanism in Kv channels. In this review we will discuss the interaction of HaTx and related spider peptides with Kv channels illustrating the properties of the binding surface of these peptides, their membrane partitioning characteristics, and will provide a working hypothesis for how the peptides inhibit gating of Kv channels. Advanced simulation results support the concept of mutual conformational changes upon peptide binding to the S3b region of the channel which will restrict movement of S4 and compromise coupling of the gating machinery to opening of the pore. PMID- 17113616 TI - Peptides inhibitors of acid-sensing ion channels. AB - Acid-sensing ion channels (ASICs) channels are proton-gated cationic channels mainly expressed in central and peripheric nervous system and related to the epithelial amiloride-sensitive Na(+) channels and to the degenerin family of ion channels. ASICs comprise four proteins forming functional channel subunits (ASIC1a, ASIC1b, ASIC2a, and ASIC3) and two proteins (ASIC2b and ASIC4) without yet known activators. Functional channels are activated by external pH variations ranging from pH(0.5) 6.8 to 4.0 and currents are characterized by either rapid kinetics of inactivation (ASIC1a, ASIC1b, ASIC3) or slow kinetics of inactivation (ASIC2a) and sometimes the presence of a plateau phase (ASIC3). ASIC1a and ASIC3, which are expressed in nociceptive neurons, have been implicated in inflammation and knockout mice studies support the role of ASIC3 in various pain processes. ASIC1a seems more related to synaptic plasticity, memory, learning and fear conditioning in the CNS. ASIC2a contributes to hearing in the cochlea, sour taste sensation, and visual transduction in the retina. The pharmacology of ASICs is limited to rather nonselective drugs such as amiloride, nonsteroid anti inflammatory drugs, and neuropeptides. Recently, two peptides, PcTx1 and APETx2, isolated from a spider and a sea anemone, have been characterized as selective and high-affinity inhibitors for ASIC1a and ASIC3 channels, respectively. PcTx1 inhibits ASIC1a homomers with an affinity of 0.7 nM (IC(50)) without any effect on ASIC1a containing heteromers and thus helped to characterize ASIC1a homomeric channels in peripheric and central neurons. PcTx1 acts as a gating modifier since it shifts the channel from the resting to an inactivated state by increasing its affinity for H(+). APETx2 is less selective since it inhibits several ASIC3 containing channels (IC(50) from 63 nM to 2 microM) and to date its mode of action is unknown. Nevertheless, APETx2 structure is related to other sea anemone peptides, which act as gating modifiers on Nav and Kv channels. PMID- 17113617 TI - Double-layer PVDF transducer and V(z) measurement system for measuring leaky Lamb waves in a piezoelectric plate. AB - This paper presents a new experimental measurement method for leaky Lamb waves propagating in a piezoelectric plate immersed in a conductive fluid. The measurement system is a low-frequency version of lens-less acoustic microscopy which has been developed based on a line-focus double-layer PVDF transducer. The transducer and its defocusing measurement system can perform V(z) measurements on a sample plate immersed in a fluid, and therefore can obtain the leaky Lamb wave velocities with high accuracy. An X-cut LiNbO(3) plate is investigated with this experimental measurement system to find out its fluid-loading effects, especially the conductive loading effects by water of various conductivities. Angular dependence of this conductive loading effect along different propagating directions on the X-cut LiNbO(3) plate is measured. It is found out the conductive loading effects are strongly dependent on the piezoelectric coupling factor. Theoretical calculations based on partial wave theory have also been carried out and compared with experimental data. Good agreements have been observed. PMID- 17113618 TI - Molecular features of new Peach Latent Mosaic Viroid variants suggest that recombination may have contributed to the evolution of this infectious RNA. AB - Nucleotide sequences of a broad range of Peach Latent Mosaic Viroid (PLMVd) variants were determined. The variants were isolated from peach, pear, and almond tree samples collected in Tunisia. Sequence analysis confirmed the high variability of PLMVd, as no less than 119 new variants were identified. Variations included new polymorphic positions, insertions of 11 to 14 nucleotides, and new mutations within the hammerhead self-cleavage motifs. We provide the first covariation-based evidence for certain stems within the proposed secondary structure. Our covariation analysis also strengthens the view that a pseudoknot closes the replication domain. On the basis of phylogenetic tree studies and informative positions, PLMVd variants are proposed to cluster into groups and subgroups likely to have resulted from recombination events. PLMVd thus emerges as a suitable viroid for retracing the evolution of an RNA genome. PMID- 17113619 TI - Diversity and evolution of West Nile virus in Illinois and the United States, 2002-2005. AB - Evolutionary analyses of West Nile virus (WNV) have been limited by uneven sampling across geographic regions and over time. In this study, an expanded data set of 68 WNV envelope gene sequences from the Midwest (Illinois) was created and combined with published sequences to investigate spatial and temporal structuring in the United States viral population. Results indicate an overall lack of geographic structure to WNV in the United States, supporting the notion of WNV as a rapidly expanding pathogen not significantly restricted in its spread by geographic distance. However, analyses of viral genetic diversity show a steady increase in WNV nucleotide-level diversity over time. Additionally, evolutionary rate calculations indicate that WNV has evolved at approximately 0.85 x 10(-3) substitutions/site/year, largely through neutral substitution and purifying selection. Overall, these results show WNV across the United States to be a panmictic viral population that is diversifying and evolving. PMID- 17113620 TI - Effect of ultrasound pretreatment in mesophilic and thermophilic anaerobic digestion with emphasis on naphthalene and pyrene removal. AB - In many anaerobic digestion processes for the treatment of the sludge produced in wastewater treatment plants, the hydrolysis of the organic matter has been identified as the rate limiting step. This study is focused on the effect of ultrasonic pretreatment of raw sewage sludge before being fed to the mesophilic and the thermophilic anaerobic digestion. From particle size reduction, COD disintegration degree and biodegradability test, 11,000kJ/kg TS was estimated as the optimal specific energy in ultrasonic pretreatment. Moreover, the use of pretreated sludge improved significantly the COD removal efficiency and biogas production in lab-scale anaerobic digesters when compared with the performance without pretreatment, specially under mesophilic conditions. During ultrasonic pretreatment, the diffusion of polycyclic aromatic hydrocarbons (PAH) compounds to the aqueous phase was stated by a reduction in the pretreated sludge micropollutants content. With sonication, naphthalene was better removed than without this pretreatment, particularly in the mesophilic digester. However, pyrene removal remained at same efficiency level with and without ultrasonic pretreatment. PMID- 17113621 TI - Development and application of a resazurin-based biomass activity test for activated sludge plant management. AB - A rapid, robust and cost-effective method of assaying the metabolic activity of the biomass of activated sludge plants would be a valuable process control tool in the wastewater treatment industry. We have developed and optimised a simple colorimetric test protocol, based on the redox dye resazurin, in which levels of reduction of the dye are proportional to cell biomass and respiration rate in both freshly sampled municipal sludges and in a surrogate activated sludge culture, Polytox. The method has been used to assess the impact of trade wastes on the activities of two municipal activated sludge populations of differing characteristics. PMID- 17113622 TI - Adsorption enhancement of laterally interacting phenol/aniline mixtures onto nonpolar adsorbents. AB - Adsorption equilibria of phenol and aniline onto nonpolar macroreticular adsorbents were investigated in single and binary-solute aqueous systems at 293 K and 313 K. All adsorption isotherms can be well represented by the Langmuir equation. Larger uptake of aniline than phenol onto all the adsorbents probably results from the higher hydrophobicity of the former compound as well as the greater electronic density of the aromatic ring of aniline. It is interestingly observed that at a relatively high loading, the total uptake of phenol and aniline in a binary system is remarkably higher than those in a single system. Such uptake difference was elucidated by the cooperative effect arising from the lateral acid-base interaction between the loaded phenol and aniline molecules. Moreover, larger average pore size of the adsorbent is found to result in a greater cooperative coefficient, as observed from the equimolar phenol/aniline adsorption system. PMID- 17113623 TI - Polycyclic aromatic hydrocarbon emissions from clinical waste incineration. AB - Since the introduction of the Environmental Protection Act in the UK, there are few reports of PAH emissions from clinical waste incinerators (CWIs) operating to improved performance standards. The main aim of this study is to determine PAH emissions from a state-of-the-art CWI focusing on the effects of reactive gases and operating variables on emissions. This was carried out by collection of stack samples over three phases of operation. At stack conditions, most PAHs are predicted to be in the vapour phase. Reactive losses of PAHs were closely correlated by rank with expected reactivities from laboratory studies. Estimates of emissions incorporating sampling losses were derived, although no correlation was found between PAH losses and the modest levels of reactive stack gases. PAH concentrations were one to two orders of magnitude lower than earlier reports from incinerators without effective air pollution control equipment (APCE). The low levels of carbon monoxide recorded were not correlated with any PAHs. This study demonstrates the impact of efficient combustion conditions and APCE on PAH emissions from a CWI. PMID- 17113624 TI - Microbial activities in soils of a former sawmill area. AB - To find out microbial metabolic functioning and toxicity in a former sawmill area, carbon dioxide evolution, methane oxidation potential, 10 hydrolytic enzyme activities, Vibrio fischeri test, fluorescein diacetate hydrolysis activity (FDA), soil pH, carbon, nitrogen and pentachlorophenol (PCP) content were measured at four sites. The area is contaminated with aged chlorophenols. Chlorophenol content of soil was analyzed with a novel HPLC-MS technique, which allowed to measure chlorophenols without derivatization. The sites had a pollution gradient from 0.5 to 15 microg PCP g dw of soil(-1). Endogenous carbon dioxide evolution, methane oxidation potential, butyrate-esterase, acetate esterase, sulphatase and aminopeptidase activities were lower at the site 2 than 3, although the site 2 and 3 had similar content of carbon and nitrogen. The soil was toxic in V. fischeri test at the site 2, which had high content of PCP (3.93+/-1.00 microg PCP g dw of soil(-1)). The results indicated that endogenous carbon dioxide evolution, methane oxidation potential, butyrate-esterase, acetate esterase, sulphatase and aminopeptidase activities were sensitive to PCP in the soil. The results indicated that alpha-glucosidase, beta-glucosidase, beta xylosidase, beta-cellobiosidase, phosphomonoesterase, N-acetyl-glucosaminidase activity and FDA hydrolysis activity were not sensitive to PCP in the soil. Soil processes involved in the cycling of carbon, nitrogen, sulphur and phosphorus were only slightly vulnerable in the former sawmill area and most sensitive microbial species were probably replaced with more tolerant ones to maintain and recover functioning of the former sawmill soils. PMID- 17113625 TI - Crystal structure analysis of enantiomerically pure (+) and (-) beta hexabromocyclododecanes. AB - The molecular structures of individual HBCD stereoisomers are not elucidated yet. Recently, we isolated 8 of the 16 possible stereoisomers from a technical HBCD mixture and tentatively assigned their relative configurations. Herein we report on the isolation of enantiomerically pure (+) and (-) beta-HBCDs, both obtained from preparative chiral-phase liquid chromatography, and we present their absolute configurations determined from X-ray diffraction analysis. The absolute configuration of (+) beta-HBCD was found to be (1S,2S,5S,6R,9S,10R), while the one of (-) beta-HBCD was assigned to (1R,2R,5R,6S,9R,10S). The given structural information allows, for the first time, the unambiguous identification of these two important HBCD stereoisomers, which are typically found in technical products at proportions of about 3-5% for each enantiomer. PMID- 17113626 TI - The redox state and activity of superoxide dismutase classes in Arabidopsis thaliana under cadmium or copper stress. AB - The redox state of glutathione and ascorbate as well as the activity of superoxide dismutase classes were determined in leaves of Arabidopsis thaliana grown for seven days in the nutrient solution containing 0, 5 and 50 microM Cd or Cu excess. A decrease in GSH/GSSG ratio was found in plants under Cd and Cu stress. In the plants exposed to Cu stress the activity of all SOD classes increased. However, in the plants treated with Cd the activity of FeSOD and MnSOD was elevated, but CuZnSOD activity was diminished in comparison with control. In these plants the activity of SOD classes was dependent on both the GSH/GSSG and AA/DHA ratios, while in those exposed to Cu excess - on the GSH/GSSG ratio. Differences were shown in the changes both in redox state and activity of SOD classes caused by the metals differing in physiochemical properties. Moreover, relationships between changes in SOD class activities and ROS levels were discussed. PMID- 17113627 TI - QSPR model of Henry's law constant for a diverse set of organic chemicals based on genetic algorithm-radial basis function network approach. AB - Six quantitative structure-property relationship (QSPR) models for a diverse set of experimental data of Henry's law constant (H) of organic chemicals under environmental condition (T=25 degrees C; water-air system) have been developed based on four different molecular descriptor sets. Three different models based on the descriptors of CODESSA (Comprehensive Descriptors for Structural and Statistical Analysis), Tsar, and Dragon software and a model based on a combined descriptor set from these packages, and in addition from HYBOT software, have been established using the stepwise regression method. The combined descriptors set model gave the best results. Furthermore, a genetic algorithm was used for descriptor selection from a combined set of descriptors, and a radial basis function network was utilized to establish a model with a low root mean square error (RMSE). The results of this study were compared with the well-known bond contribution and group contribution methods. The group contribution method failed to predict Henry's law constant of 170 from all 940 compounds in the data-set. RMSEs of 0.693, 0.798, and 0.564 were achieved for bond contribution, group contribution and the best QSPR model of this study, respectively, based on logarithm of H. Analysis of different QSPR models showed that hydrogen bonding between the organic solute and water as a solvent has the greatest influence on this partitioning phenomenon. PMID- 17113628 TI - Regulatory networks: linking microarray data to systems biology. AB - Gene regulation and aging are intrinsically linked and these links often reach directly to transcription factors and their actions in gene regulation. However, it is very difficult to follow all the individual directions such factors can affect. Therefore, the opposite approach became more popular recently, i.e. observing the endpoints of all these actions. Microarrays are the preferred technology to monitor large-scale changes in transcripts across whole genomes. The trade-off for being able to survey whole genome transcriptomes is that the results are mere observations, which do not directly reveal the underlying mechanisms that represent the real link to transcription factors and their actions. Fortunately, a combination of knowledge mining (including but not restricted to literature mining) with genomics analyses can be harnessed to elucidate at least some of the regulatory networks orchestrating the transcriptional changes observed by microarray experiments. Thus, a considerable part of the functional system structure of cells and organisms can be revealed, which is a pivotal prerequisite for any meaningful systems biology approach towards aging related phenotypes. PMID- 17113629 TI - GAGEC1, a cancer/testis associated antigen family member, is a target of TGF beta1 in age-related prostatic disease. AB - Transforming growth factor beta (TGF-beta) is a multi-functional cytokine that plays a fundamental role during embryonic development and tissue homeostasis in metazoans. Changes in TGF-beta signalling are implicated in prostate cancer (PCa) and benign prostatic hyperplasia (BPH), two of the most common diseases affecting ageing males. GAGEC1 belongs to the GAGE-related family of cancer/testis associated antigens and in males is expressed only in prostate and testis. Previous reports demonstrate that GAGEC1 is up-regulated in symptomatic BPH and PCa. We demonstrate GAGEC1 up-regulation by TGF-beta1 in primary prostatic stromal and epithelial cells. Our data suggest that disease-associated increases in TGF-beta1 may account for the increase in GAGEC1 expression in BPH and PCa. Given its restricted spatial expression in males, GAGEC1 represents a promising target for therapeutic intervention of BPH and PCa. PMID- 17113630 TI - Use of dogs as indicators of metal exposure in rural and urban habitats in NW Spain. AB - Many different species have been used in environmental biomonitoring studies in diverse habitats including forest, farmland, and urban and sub-urban areas. However, there is little information on domestic animals living in rural or urban habitats and exposed to the same pollutants as the human population. In this connection, pets could prove to be good indicators of human metal exposure since they closely share the same environment as their owners, and are therefore exposed, at least in part, to the same pollutants. The present study investigated toxic metal exposure in dogs in NW Spain and compared metal exposures between dogs from rural and urban habitats, considering the influence of diet, sex and age. Samples of liver and kidney from 57 male and female dogs, aged between 6 months and 18 years, were collected after euthanasia at veterinary clinics. Samples were acid-digested and metal concentrations determined by ICP-MS. Geometric mean concentrations of metals in the liver and kidney (microg/kg wet weight) were 12.6 and 15.9 for arsenic, 58.0 and 175 for cadmium, 32.7 and 53.4 for mercury, and 57.7 and 23.1 respectively. Hepatic lead concentrations were significantly higher (p<0.05) in dogs fed commercial diets than dogs fed home made feed (32%) or a mixture of commercial and home-made feeds (95%). Mercury concentrations in the kidney were significantly higher (3-fold, p<0.05) in dogs from urban areas than in dogs from rural areas. Cadmium levels in kidney were significantly higher (p<0.05) in females (67%) and increased with age (p<0.001). Although no human samples were obtained in this study and no direct correlations between dogs and human metal exposure have been conducted, given our results pets could be suggested as surrogate indicators of human metal exposure. PMID- 17113631 TI - Arsenic uptake by common marsh fern Thelypteris palustris and its potential for phytoremediation. AB - Hydroponic and soil cultivations of Thelypteris palustris, the common marsh fern, were used to investigate its potential for use in phytoremediation of arsenic (As) contaminated water or soil. ICP-MS analyses indicate that both roots and fronds accumulated arsenic in levels up to 100 times the concentration of treatment solutions of 250 microg/L and 500 mug/L arsenic, but values varied widely and there was no significant difference in concentrations in fronds between the control (no arsenic) and treatments. Plants exposed to 500 microg/L exhibited necrosis in their fronds, suggesting that Thelypteris palustris is not a good candidate for phyotoremediation of arsenic-contaminated sites. PMID- 17113632 TI - Cytokine gene variants and venous thrombotic risk in the BRATROS (BRAZILIAN THROMBOSIS STUDY). AB - INTRODUCTION: Venous thrombosis (VT) and inflammation are two closely related entities. In the present investigation we assessed whether there is a relation between genetic modifiers of the inflammatory response and the risk of VT. MATERIALS AND METHODS: 420 consecutive and unrelated patients with an objective diagnosis of deep VT and 420 matched controls were investigated. The frequencies of the following gene polymorphisms were determined in all subjects: TNF-alpha 308 G/A, LT-alpha+252 A/G, IL-6-174 G/C, IL1-ra 86 bp VNTR, IL-10-1082 A/G and CD 31 125 C/G. RESULTS: Overall odds ratio (OR) for VT related to TNF-alpha-308 G/A, LT-alpha+252 A/G, IL-6-174 G/C, A1 allele (4 bp repeat) of the IL1-ra 86 bp VNTR, IL-10-1082 A/G and CD-31 125 C/G were respectively: 1.0 (CI95: 0.8-1.5), 1.3 (CI95: 1.0-1.7), 1.1 (CI95: 0.9-1.5), 1.6 (CI95: 1-2.5), 1.2 (CI95: 0.8-1.7) and 0.8 (CI95: 0.6-1.1). A possible interaction between polymorphisms was observed only for the co-inheritance of the mutant alleles of the LT-alpha+252 A/G and IL 10-1082 G/A polymorphisms (OR=2; CI95: 1.1-3.8). The risk of VT conferred by factor V Leiden and FII G20210A was not substantially altered by co-inheritance with any of the cytokine gene polymorphisms. CONCLUSIONS: Cytokine gene polymorphisms here investigated did not significantly influence venous thrombotic risk. PMID- 17113633 TI - In vitro induction of differentiation by retinoic acid in an immortalized olfactory neuronal cell line. AB - In this study, we used a neuronal cell line generated by transfection of rat olfactory epithelium with immortalizing recombinant oncogene E1A of adenovirus-2. The resulting 13.S.1.24 line of transformed cells expressed an antigenic phenotype of olfactory neuronal progenitors. Time-dependency assessments over 1 week of treatment indicated that apoptosis and differentiation induced by retinoic acid (RA) were concomitant. Indeed, RA altered the cell proliferation rate, but it also stimulated differentiation of surviving 13.S.1.24 cells into bipolar olfactory marker protein-immunoreactive neurons. To characterize the nature of the cells we used immunocytochemistry, optical imaging, scanning electron microscopy and atomic force microscopy. PMID- 17113634 TI - The melanocortin agonist, melanotan II, enhances proceptive sexual behaviors in the female rat. AB - Melanocortins have been reported to play a role in the control of both male and female sexual behavior. The present study examined the effects of melanotan-II (MT-II), a cyclic peptide analogue of alpha-melanocyte stimulating hormone on appetitive and consummatory aspects of female sexual behavior, including aspects of sexual proceptivity (solicitations, hops and darts, ear wiggling, pacing) and receptivity (lordosis). One group of ovariectomized Long-Evans rats (n=7) was primed subcutaneously with estradiol benzoate (EB) and progesterone (P) (10 microg and 500 microg respectively) and another group (n=7) with EB (10 microg) and oil (EB alone). Paced mating tests were performed with sexually experienced males in unilevel chambers, which were bisected by a Plexiglas divider containing three holes, through which only the female could pass. MT-II (1 and 3 mg/kg) or saline was injected intravenously 10 min before each 30-min paced mating test. Each female received the 3 treatments. In females primed with EB+P both doses of MT-II increased the number of hops and darts and ear wiggling significantly, but did not alter pacing or lordosis. With EB alone, no effect of MT-II was observed on any of the parameters measured. These results suggest that P can interact with MT-II to increase proceptive behaviors. Because hops and darts are essentially solicitations, made in close proximity to the male, that indicate a desire on the part of females to receive mounts and intromissions, these data suggest that activation of melanocortin receptors may represent a promising mode of action for the treatment of women with hypoactive sexual desire. PMID- 17113635 TI - Effects of solvent drying time on micro-shear bond strength and mechanical properties of two self-etching adhesive systems. AB - OBJECTIVES: The all-in-one adhesives are simplified forms of two-step self etching adhesive systems that must be air dried to remove solvent and water before curing. It was investigated whether those two systems perform equally well and if their performance is affected by air-drying of the solvent containing agent. METHODS: Two adhesive systems (both by Kuraray Medical) were evaluated; Clearfil Tri-S bond (TS) and Clearfil SE bond (SE). Micro-shear bond strengths to human dentin after solvent air-drying times of 2, 5 or 10 s for each group were measured (n=10). The indentation creep and hardness of the bonding layer were also determined for each group. RESULTS: The lowest micro-shear bond strength, nano-indentation hardness and creep stress exponents were obtained for 2 s air dried specimens of each material. After 10 s air blowing, SE showed superior properties compared to TS groups (p<0.05). SIGNIFICANCE: When properly handled, two step self-etching material performs better than the all-in-one adhesive. Air drying is a crucial step in the application of solvent containing adhesives and may affect the overall clinical performance of them, through changes in the bond strength and altering nano-scale mechanical properties. PMID- 17113636 TI - Increased angiogenesis and blood vessel maturation in acellular collagen-heparin scaffolds containing both FGF2 and VEGF. AB - An important issue in tissue engineering is the vascularisation of the implanted construct, which often takes several weeks. In vivo, the growth factors VEGF and FGF2 show a combined effect on both angiogenesis and maturation of blood vessels. Therefore, we hypothesise that the addition of these growth factors to an acellular construct increases blood vessel formation and maturation. To systematically evaluate the contribution of each scaffold component with respect to tissue response and in particular to blood vessel formation, five porous scaffolds were prepared and characterised, viz.: collagen, collagen with heparin, and collagen with heparin plus one or two growth factors (rrFGF2 and rrVEGF). Scaffolds were subcutaneously implanted in 3 months old Wistar rats. Of all scaffolds tested, the one with a combination of growth factors displayed the highest density of blood vessels (type IV collagen) and most mature blood vessels (smooth muscle actin). In addition, no hypoxic cells were found in this scaffold at day 7 and 21 (hypoxia inducible factor 1-alpha). These results indicate that the addition of both FGF2 and VEGF to an acellular construct enhances an early mature vasculature. This opens prospects for (acellular) tissue-engineered constructs in conditions as ischaemic heart disease or diabetic ulcers. PMID- 17113637 TI - PEGylation of microspheres for therapeutic embolization: preparation, characterization and biological performance evaluation. AB - In this study, microspheres designed for embolization, defined as GF2000 Trisacryl MS (GF-MS) and DEAE-Trisacryl MS (DEAE-MS), were originally PEGylated using (3-amino propyl) triethoxy silane as coupling agent. Indomethacin was loaded into both PEGylated and non-PEGylated DEAE-MS, displaying ion-exchange ability, through a batch process with a respective capacity of 1.2 and 0.25 g/g. The morphology of naked and PEGylated MS was evaluated by scanning electron microscopy (SEM). Both micosphere resins surface looked like orange skin, although DEAE-MS showed a slightly rougher surface due to the copolymerization process. PEGylated microspheres have a most likely swelling surface owing to the presence of PEG hydrophilic chains. The mean diameters were of about 66 and 60 microm for GF-MS and DEAE-MS, respectively. Data obtained for PEGylated MS by Fourier Transform Infrared spectroscopy (FTIR) confirmed that microspheres were successfully PEGylated. Finally, complement activation in vitro was performed to evaluate the activating capacity of different microspheres. Both PEGylated GF-MS and DEAE-MS activated the complement system of about 33% less than their corresponding naked microspheres, while loading PEGylated DEAE-MS with indomethacin almost suppressed complement activation. This inhibiting role implies that PEGylation as well as loading the microspheres with anti inflammatory drug has a compact effect on the interaction of microspheres with blood proteins. PMID- 17113638 TI - Prognostic signature of ALL blasts at diagnosis: what can we really find? PMID- 17113639 TI - Match unrelated bone marrow transplantation in a case of high risk myelodysplastic syndrome treated with azacitidine and concomitant 1alpha-25 dihydroxyvitamin D3, as differentiating agent. PMID- 17113640 TI - The discovery of endothelial progenitor cells. An historical review. AB - Although the earliest sites of hematopoietic cell and endothelial cell differentiation in the yolk sac blood islands were identified about 100 years ago, cells with hemangioblast properties have not yet been identified in vivo. Endothelial cells differentiate from angioblasts in the embryo and from endothelial progenitor cells, mesoangioblasts and multipotent adult progenitor cells in the adult bone marrow. Endothelial progenitor cells (EPC) were initially described by Asahara et al. [Asahara T, Murohara T, Sullivan A, et al. Isolation of putative progenitor endothelial cells for angiogenesis. Science 1997;275:964 7.], and the past few years have seen a rapid expansion of our knowledge of EPC biology. Prior to the discovery of this cell type, new vessel formation was believed to occur to proliferation of existing endothelial cells. These findings have overturned the previous dogma that vasculogenesis can only occur during embryogenesis. Questions persist regarding their functional characteristics, as well as the precise panel of cell surface markers that define this cell population. PMID- 17113642 TI - Correlates of problem recognition and intentions to change among caregivers of abused and neglected children. AB - OBJECTIVE: To identify individual, family, and caseworker characteristics associated with problem recognition (PR) and intentions to change (ITC) in a sample of caregivers who received in-home child welfare services following substantiated reports of child abuse or neglect. METHODS: Caregivers were interviewed at 4 weeks, 16 weeks, and 1 year after referral for in-home services. In these interviews, the University of Rhode Island Change Assessment scale assessed PR and ITC in relation to caregiving practices. Additional data were obtained from administrative records and surveys of in-home services caseworkers. We used growth models to identify caregiver, family, and caseworker characteristics associated with initial levels of PR and ITC, and with changes in PR and ITC over time. RESULTS: Contrary to expectations, there were no overall increases in PR and ITC during the first 4 months of in-home services. PR and ITC scores fluctuated over time, in relation to some external events, case characteristics, and caseworker characteristics; however, we were able to account for small portions of the variance in PR and ITC. Controlling for social desirability bias and other variables in the analysis, negative life events, lack of network support, and the severity of caregiver depression were associated with greater PR; housing problems were associated with lower PR. Caregiver depression and age were associated with ITC. Caregivers whose children had been removed from their homes showed significant increases in ITC in the first few months of in home services. The duration of caseworkers' child welfare experience predicted increases in their clients' PR in the first 4 months and more frequent contacts with an experienced caseworker predicted small, but significant increases in ITC over time. CONCLUSIONS: PR and ITC are associated with somewhat different case characteristics and may be affected by caseworkers' experience. PMID- 17113643 TI - Child protection and welfare reform. PMID- 17113644 TI - Evolving a theoretical model of child safety in maltreating families. PMID- 17113645 TI - Genotyping of Edwardsiella tarda isolated from freshwater fish culture system. AB - The applicability of PCR-RFLP of 16S rDNA and conventional phenotypic methods for differentiation of Edwardsiella tarda associated in freshwater fish culture system was studied. In this study, by conventional biochemical tests and antibiotic resistant patterns 2 and 14 groups were obtained. But these methods failed to discriminate the isolates habitat wise. However, PCR-RFLP of 16S rDNA was found to be specific to detect habitat-specific isolates. All the fish isolates belonging to particular genotypes were found only in fish, not in water or sediment. Some of the genotypes were exclusively present in water and sediment. This study indicates the prevalence of site-specific genotypes in freshwater ecosystems. Molecular method is found to be superior to discriminate the E. tarda habitat wise to conventional typing methods. PMID- 17113646 TI - Association between tofu intake and serum polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) in the elderly Taiwanese. AB - Age and gender are related to human dietary habits, which affect serum levels of dioxin-like compounds via body fat content. This study examined whether different dietary patterns and body fat content were associated with serum polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F) concentrations for different age groups of Taiwanese subjects. In total, 1165 volunteers completed a semi quantitative food-consumption frequency questionnaire for serum PCDD/Fs analysis. In the elderly (age, >45 years), a significant positive multivariate-adjusted association was identified between serum PCDD/F levels and sea fish intake. Additionally, consumption of tofu was negatively correlated with serum PCDD/F levels. This study also found a negative correlation between tofu intake and body fat content in the elderly. It seems that when elderly people ingested considerable amounts of soybean protein, such as tofu, they typically had low serum PCDD/Fs levels in relation to reduced body fat content and induction of metabolic enzymes. The relationship between tofu intake, metabolic enzymes, and serum PCDD/F accumulations warrants further investigated previously to recommend how to prevent PCDD/F accumulations via intake of soybean products. PMID- 17113647 TI - A member of the cathelicidin family of antimicrobial peptides is produced in the upper airway of the chinchilla and its mRNA expression is altered by common viral and bacterial co-pathogens of otitis media. AB - Cationic antimicrobial peptides (AMPs), a component of the innate immune system, play a major role in defense of mucosal surfaces against a wide spectrum of microorganisms such as viral and bacterial co-pathogens of the polymicrobial disease otitis media (OM). To further understand the role of AMPs in OM, we cloned a cDNA encoding a cathelicidin homolog (cCRAMP) from upper respiratory tract (URT) mucosae of the chinchilla, the predominant host used to model experimental OM. Recombinant cCRAMP exhibited alpha-helical secondary structure and killed the three main bacterial pathogens of OM. In situ hybridization showed cCRAMP mRNA production in epithelium of the chinchilla Eustachian tube and RT-PCR was used to amplify cCRAMP mRNA from several other tissues of the chinchilla URT. Quantitative RT-PCR analysis of chinchilla middle ear epithelial cells (CMEEs) incubated with either viral (influenza A virus, adenovirus, or RSV) or bacterial (nontypeable H. influenzae, M. catarrhalis, or S. pneumoniae) pathogens associated with OM demonstrated distinct microbe-specific patterns of altered expression. Collectively, these data showed that viruses and bacteria modulate AMP messages in the URT, which likely contributes to the disease course of OM. PMID- 17113649 TI - Fixed point analysis of nystagmus. AB - Motor disorders frequently contain a rhythmic component, but the associated oscillations are not usually precisely periodic. This lack of strict periodicity can make it difficult to identify the effects of experimental manipulations on the oscillation. In this report, we describe the application of a numerical technique for identifying fixed points of a nonlinear map to the recovery of underlying periodicities of the eye movement disorder of nystagmus. The technique is illustrated by application to two different types of nystagmus. In addition we use a local analysis of the behaviour at the fixed points to distinguish between different bifurcations in the two examples with changes in gaze angle. We conclude that the technique reveals consistent effects of experimental manipulations, which may be useful for quantitative characterisation of experimental and therapeutic manipulations of motor disorders. PMID- 17113648 TI - Upregulation of vitamin D binding protein (Gc-globulin) binding sites during neutrophil activation from a latent reservoir in azurophil granules. AB - Vitamin D binding protein (DBP) is a multifunctional plasma transport protein that is also found on the surface of many cell types. Cell surface DBP significantly enhances chemotactic activity of complement (C) peptides C5a and C5a des Arg. However, both DBP binding and C5a chemotaxis enhancement can vary among neutrophil donors. To test if activation during cell purification is responsible for this variability, neutrophils were isolated using both standard and lipopolysaccharide (LPS)-free protocols. Cells isolated by the LPS-free method had no DBP-enhanced chemotaxis to C5a or DBP binding to plasma membranes. Moreover, neutrophils treated with LPS bound more avidity to immobilized DBP than sham-treated cells. Subcellular fractionation of neutrophils (standard protocol) revealed a heavy plasma membrane (HM) band that contained components of light plasma membranes and all three granules. The HM band possessed most of the DBP binding activity (58%), and activation of cells with ionomycin greatly increased DBP binding to HM. Azurophil granules contained 33% of the total DBP binding sites and there was a highly significant positive correlation (r=0.988) between release of the granule marker myeloperoxidase and DBP binding. These results indicate that fusion of granules with the plasma membrane forms HM that contains DBP binding sites. PMID- 17113650 TI - Alteration of cardiac autonomic functions in patients with major depression: a study using heart rate variability measures. AB - BACKGROUND: Depression is associated with greater cardiac morbidity and mortality. One of the contributory factors for this may be altered cardiac autonomic activity in depression. However, cardiac autonomic involvement in depression remains controversial because of methodological issues. In this study, alteration of cardiac autonomic functions was studied in drug-naive patients with major depression without co-morbidity. Heart rate variability, a sensitive measure of neurocardiac autonomic regulation was used in addition to conventional methods of measuring cardiac autonomic functions. METHODS: We recruited 40 patients suffering from major depression, diagnosed based on DSM-IV-TR criteria. Their cardiac autonomic functions were measured using both conventional and heart rate variability measures. These were compared with those of age- and gender matched healthy controls. RESULTS: Patients with major depression showed significantly lesser Valsalva ratio, maximum/minimum ratio and greater sympathovagal balance than healthy controls indicating decreased parasympathetic and increased sympathetic activity. CONCLUSIONS: Depression is associated with alteration of cardiac autonomic tone towards decreased parasympathetic activity and an increased sympathetic activity. It is possible that a common neurobiological dysfunction contributes to both depression and cardiac autonomic changes in the illness. PMID- 17113652 TI - High anxiety and migraine are associated with the s allele of the 5HTTLPR gene polymorphism. AB - The 5HTTLPR polymorphism of the serotonin transporter gene has been associated with anxiety disorders and also migraine, suggesting a common etiological background of these disorders. This association is further supported by the high comorbidity of these disorders. In our study Spielberger's State-Trait Anxiety Inventory and the 5HTTLPR genotype were investigated in a cohort of 97 psychiatrically healthy females also including 45 migraineurs. Higher state anxiety scores were significantly associated with the s allele either in the whole sample or when the group was separated into migraineurs and non migraineurs. Migraineurs also had a significantly higher frequency of the s allele. Our results indicate that even in a healthy population the s allele is associated with a high anxiety endophenotype. The association of migraine with anxiety may be explained by the higher rate of individuals carrying the s allele among migraineurs. PMID- 17113651 TI - Early developmental characteristics and features of major depressive disorder among child psychiatric patients in Hungary. AB - OBJECTIVE: We investigate the relations of early atypical characteristics (perinatal problems, developmental delay, and difficult temperament) and onset age (as well as severity of) first major depressive disorder (MDD) and first internalizing disorder in a clinical sample of depressed children in Hungary. METHOD: Participants were 371 children (ages 7-14) with MDD, and their biological mothers, recruited through multiple clinical sites. Diagnoses (via DSM-IV criteria) and onset dates of disorders were finalized "best estimate" psychiatrists, and based on multiple information sources. Mothers provided developmental data in a structured interview. RESULTS: Difficult temperament predicted earlier onset of MDD and first internalizing disorder, but its effect was ameliorated if the family was intact during early childhood. Further, the importance of difficult temperament decreased as a function of time. Perinatal problems and developmental delay did not impact onset ages of disorders, and none of the early childhood characteristics associated with MDD episode severity. CONCLUSIONS: Children with MDD may have added disadvantage of earlier onset if they had a difficult temperament in infancy. Because early temperament mirrors physiological reactivity and regulatory capacity, it can affect various areas of functioning related to psychopathology. Early caregiver stability may attenuate some adverse effects of difficult infant temperament. PMID- 17113653 TI - Lactoferrin ameliorates symptoms of experimental encephalomyelitis in Lewis rats. AB - Lactoferrin (LF) is a multifunctional protein present in secretory fluids of mammals and circulating neutrophils. Beside anti-inflammatory properties, LF was found to inhibit some autoimmune disorders. In this investigation we studied effects of oral administration of LF on experimental autoimmune encephalomyelitis (EAE) in Lewis rats. LF was given in drinking water as 0.25% solution beginning the day of elicitation of EAE or with a seven-day delay. The effects of LF were evaluated by the following criteria: clinical score, lymph node cell number, serum cytokine levels and histopathological changes. We found that LF treatment led to a significant acceleration of the recovery process, particularly on days 16-18 following elicitation of EAE. The delayed administration of LF was less effective in reducing the score of EAE. In addition, cell number of the inguinal lymph nodes of untreated EAE rats, almost 3 times higher as compared with control, naive rats, was normalized by LF treatment. Furthermore, LF decreased elevated serum concentrations of tumor necrosis factor alpha and transforming growth factor beta. The histological analysis of the spinal cord revealed reduction in the number and size of inflammatory foci in LF-treated rats. In summary, treatment of EAE Lewis rats with LF reduced the clinical symptoms and accelerated the recovery of animals. PMID- 17113654 TI - Inhibitory effect of pasireotide and octreotide on lymphocyte activation. AB - Somatostatin (SST) regulates the function of the central and peripheral nervous system, the endocrine and exocrine organs, as well as the vascular and immune system. These actions are mediated by five specific membrane somatostatin receptors. This study compares the effects on human lymphocytes of two long acting somatostatin analogues that have different receptor affinity: octreotide and pasireotide. Both analogues have an antiproliferative effect on human lymphocyte proliferation, but they act at different concentration and, while octreotide enhances IL10 and inhibits gamma IFN pasireotide inhibits IL2 and gamma IFN. In both sets of experiment the different behaviour of the two analogues could be due to their different affinity to the SSTR subtypes. Finally this study suggest that the growth inhibitory action of somatostatin analogues is an apoptotic phenomenon and it can be mediated by SSTR2a, in the case of octreotide, and by SSTR3 when pasireotide is used or it can be mediated by the heterodimerization of the two receptor. PMID- 17113655 TI - Lasting changes in neuronal activation patterns in select forebrain regions of aggressive, adolescent anabolic/androgenic steroid-treated hamsters. AB - Repeated exposure to anabolic/androgenic steroids (AAS) during adolescence stimulates high levels of offensive aggression in Syrian hamsters. The current study investigated whether adolescent AAS exposure activated neurons in areas of hamster forebrain implicated in aggressive behavior by examining the expression of FOS, i.e., the protein product of the immediate early gene c-fos shown to be a reliably sensitive marker of neuronal activation. Adolescent AAS-treated hamsters and sesame oil-treated littermates were scored for offensive aggression and then sacrificed 1 day later and examined for the number of FOS immunoreactive (FOS-ir) cells in regions of the hamster forebrain important for aggression control. When compared with non-aggressive, oil-treated controls, aggressive AAS-treated hamsters showed persistent increases in the number of FOS-ir cells in select aggression regions, namely the anterior hypothalamus and lateral septum. However, no differences in FOS-ir cells were found in other areas implicated in aggression such as the ventrolateral hypothalamus, bed nucleus of the stria terminals, central and/or medial amygdala or in non-aggression areas, such as the samatosensory cortex and the suprachiasmatic nucleus. These results suggest that adolescent AAS exposure may constitutively activate neurons in select forebrain areas critical for the regulation of aggression in hamsters. A model for how persistent activation of neurons in one of these brain regions (i.e., the anterior hypothalamus) may facilitate the development of the aggressive phenotype in adolescent-AAS exposed animals is presented. PMID- 17113656 TI - The effects of forced exercise on hippocampal plasticity in the rat: A comparison of LTP, spatial- and non-spatial learning. AB - Physical activity may have the potential to improve cognitive function. Here we show that forced treadmill-running results in selective improvements in hippocampal plasticity. Rats that underwent exercise training demonstrated enhanced expression of long-term potentiation in dentate gyrus and enhanced object recognition learning. Spatial learning in the Morris watermaze was unaffected by exercise. These changes were associated with an increase in expression of brain-derived neurotrophic factor in the dentate gyrus. PMID- 17113657 TI - The trial-spacing effect in olfactory patterning discriminations in honeybees. AB - Harnessed bees conditioned to associate odors and sucrose reward learn to discriminate between olfactory mixtures and their odor components in negative (NP: A+, B+, AB-) and positive (PP: A-, B-, AB+) patterning experiments. They thus extend the proboscis to the reinforced (CS+) but not to the non-reinforced (CS-) stimuli. Using the same protocol, we studied whether or not trials, which are spaced in time, are more effective in supporting patterning discrimination than massed trials which succeed fast to each other ('trial-spacing effect'). Training followed a NP (4 A+, 4 B+, 8 AB-) or a PP (4 A-, 4 B-, 8 AB+) schedule, with a 1:1 ratio between CS+ and CS- trials (8 CS+ and 8 CS- trials). ITIs of 1, 3, 5 and 8min were used in both tasks. Increasing ITI resulted in better differentiation between reinforced and non-reinforced CSs in both NP and PP tasks. However, whereas only the longest ITI of 8min allowed discrimination in NP, PP could already be solved with an ITI of 5min. This difference might be due to the fact that NP, but not PP, would require the formation of a unique cue and thus longer processing times. We thus show that the trial-spacing effect, previously demonstrated for single stimulus conditioning, also determines performance in patterning tasks in which three different stimuli (A, B, AB) alternate so that elements have to be discriminated from their compound. PMID- 17113658 TI - Copper-induced oxidative stress and responses of antioxidants and phytochelatins in Hydrilla verticillata (L.f.) Royle. AB - Copper, though essential, is potentially toxic heavy metal at supraoptimal level and has widespread contamination. The present investigation was carried out to study the responses induced by lower as well as higher doses of copper (0.1-25 microM) in an aquatic macrophyte, Hydrilla verticillata (L.f.) Royle for a period of 1-7 days. The plants accumulated copper in high amount with a maximum of 770 microg g(-1) dw on day 7 at 25 microM. Biomass and photosynthetic pigments showed less alteration up to 1 microM while at higher concentrations, significant decline occurred. Malondialdehyde (MDA) content and electrical conductivity (EC) also showed sharp increase at higher concentrations indicating oxidative stress. In response to copper exposure, plants showed significant induction of proteins and enzymes like superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), catalase (CAT) and glutathione reductase (GR), however, only up to moderate exposures. Total non-protein thiols (NP-SH) and cysteine levels increased significantly up to 5 microM copper exposure while at 25 microM, their level declined drastically. Reduced glutathione (GSH) showed decrease at all concentrations while oxidized glutathione (GSSG) simultaneously increased. Phytochelatins (PCs) were also induced significantly at studied concentrations of 1 and 5 microM on day 4 in comparison to control. However, copper chelation depicted by PC-SH to copper ratio was found to be low (6.5% at 1 microM and 2.4% at 5 microM) suggesting that PCs play only a part in integrated mechanisms of copper homeostasis and detoxification. Tolerant response of plants to moderate copper exposures and high accumulation potential warrants their suitability for remediation of moderately copper polluted water bodies. PMID- 17113659 TI - The pancreatic polypeptide family and the migrating motor complex of the rat: differential effects in the duodenum and jejunum. AB - AIM: To investigate the effects of members of the pancreatic polypeptide family on migrating myoelectric complexes in rats in vivo. METHODS: Rats were supplied with bipolar electrodes at 5 (duodenum), 15 and 25 cm (jejunum) distal to pylorus for electromyography. The natural ligands neuropeptide Y, pancreatic polypeptide, peptide YY1-36 and peptide YY3-36 were infused IV at doses of 0.5-400 pmol kg(-1) min(-1). The mechanisms of action were studied after pre-treatment with N(omega) nitro-L-arginine (L-NNA) 1 mg kg(-1), guanethidine 3 mg kg(-1) and in bilaterally vagotomized animals. RESULTS: PP inhibited myoelectrical activity dose dependently in both the duodenum (ED50 5.8 pmol kg(-1) min(-1)) and jejunum (2.6 pmol kg(-1) min(-1)). PYY1-36 and PYY3-36 also had inhibitory effect in the jejunum (4.4 and 130 pmol kg(-1) min(-1), respectively). PYY1-36 had no significant effect in the duodenum, whereas PYY3-36 stimulated myoelectrical activity at the highest doses. NPY was without effect. In the jejunum neither L NNA, guanethidine or vagotomy had any significant influence on the inhibitory effects of PP, PYY1-36 and PYY3-36. In the duodenum, the effect of PP was inhibited by guanethidine, but not L-NNA or vagotomy. The stimulatory effect of PYY3-36 in the duodenum was blocked by L-NNA and vagotomy, whereas guanethidine was without effect. CONCLUSION: Peptides of the PP family modulate small bowel motility differentially. Whereas their general effect is inhibitory in the jejunum, the mixing duodenal compartment is stimulated by PYY3-36, suggested to reflect receptor distribution distinction in the gut. This implicates distribution of distinct receptors in the gut being activated by either peptide. PMID- 17113660 TI - Aging influences the level and functions of fasting plasma ghrelin levels: the POWIRS-Study. AB - OBJECTIVE: Ghrelin, known for its orexigenic activity, also have functions such as vasodilation and a growth hormone releasing action. It is uncertain whether these functions change with increasing age. This study aimed to determine whether ghrelin levels differ between young and older women with different levels of obesity; and secondly whether the associations of ghrelin with metabolic syndrome (MS) components, adipocytokines, coagulation factors, and cortisol change with increasing age. METHODS AND RESULTS: Caucasian women (N=107) were divided into young (19-29 years) and older groups (30-56 years). Fasting ghrelin, leptin, adiponectin, glucose, insulin, cortisol, fibrinogen and plasminogen activator inhibitor-1 (PAI-1) levels were determined. Blood pressure (BP), body mass index and waist circumferences were measured. Older lean women showed lower levels of ghrelin (p<0.05) than young lean women, with no differences regarding BP, obesity, lipids, adipokines or insulin resistance (IR). Ghrelin levels of older women remained constant with increasing obesity, but younger women showed significantly reduced ghrelin levels in obese groups. Only younger women showed significant correlations between ghrelin and leptin, adiponectin, fibrinogen and PAI-1 (adjusted for age, obesity and menstrual phase), whereas both age groups showed significant correlations with IR. In younger women factor analysis grouped ghrelin with coagulation factors and all MS components. In older women ghrelin was absent from the MS cluster, but was associated with lower BP, cortisol and IR. CONCLUSIONS: Ghrelin levels were not significantly elevated in lean older women, and did not change with increased obesity in older women--as were observed in younger women. The functions of ghrelin also seem to change with increased age since only in young women ghrelin was associated with obesity, coagulation factors and leptin. PMID- 17113661 TI - Exceptional survival: double inlet left ventricle presenting with aortic dissection. AB - Adult survival with unoperated univentricular heart is unusual, particularly into the latter decades. Survival is most common in balanced circulations, such as double inlet left ventricle with moderate pulmonary stenosis. We describe a woman who presented with an aortic dissection and previously unrecognized univentricular heart. PMID- 17113662 TI - Lone and secondary nonvalvular atrial fibrillation: role of a genetic susceptibility. AB - BACKGROUND: An involvement of the renin angiotensin system in atrial fibrillation (AF) has been hypothesized, and ACE DD genotype has been suggested to influence the predisposition to AF. The aim of this study was to investigate the role of the ACE I/D polymorphism in relation to the different clinical forms of AF, lone and secondary nonvalvular atrial fibrillation (NVAF). METHODS: 510 consecutive patients with documented NVAF (106 patients had lone, and 404 secondary NVAF), and 520 controls with a negative history of cardiovascular disease have been studied. RESULTS: A significant difference in allele frequency between lone and secondary NVAF (p=0.002) has been found. The ACE D allele was associated with the predisposition to lone NVAF under a dominant, recessive and additive model, both at univariate and multivariate analysis, after adjustment for age and gender (multivariate analysis: dominant OR=2.87, p=0.02; recessive OR=2.01, p=0.003; additive OR=4.47, p<0.0001). ACE D allele was significantly associated with secondary NVAF at both univariate and multivariate analysis under a recessive and additive, but not dominant, model (multivariate analysis: recessive OR=1.89, p=0.001; additive OR=2.50, p<0.0001). CONCLUSIONS: This study highlights the role of ACE gene in predisposing to both lone and secondary NVAF, further contributing to penetrate the genetic mechanisms responsible for this complex disease. The clinical relevance of our results may be related to the possible characterization of subjects predisposed to NVAF in the absence of traditional risk factors, and to the use of ACE-inhibitors therapy able to improve the arrhythmogenic substrate. PMID- 17113663 TI - Catastrophic antiphospholipid syndrome with heart involvement: diagnostic utility of the cardiac MRI. PMID- 17113664 TI - Biotechnological exploitation of bacteriophage research. AB - The experimentally amenable nature of phage and their use in testing fundamental biological questions have meant that phage research has had a profound effect on modern molecular biology. Phage research has also fuelled multiple biotechnological developments. For example, phage display has recently been harnessed in a multidisciplinary approach for the generation of novel nanotechnologies. In addition, with the emerging threat of antibiotic-resistant bacterial infections, phage have begun to provide technologies to combat these problems. Finally, recent data acquired from genome sequencing and advances in phage biology research have aided the development of phage-derived bacterial detection and treatment strategies in addition to methods to control the detrimental effects of phage in industry. Here, we examine the promising uses of phage in these important areas of biotechnology. PMID- 17113665 TI - EU biotech crop regulations and environmental risk: a case of the emperor's new clothes? AB - European Union Commissioner for the Environment Stavros Dimas recently hailed 'upgraded' non-genetically modified (GM) crops as an alternative to GM crops. A comparative analysis of the environmental risks associated with such non-GM herbicide-resistant crops and GM herbicide-resistant crops is presented here. The analysis highlights serious weaknesses in the European Union (EU) regulatory framework, and the contradictory policy of the EU Commission on the precautionary principle is also shown. The continued political stance of ignoring these regulatory and policy inconsistencies is examined and found to be flawed. It is postulated that, even in the face of these flaws and coupled with recent statements from the UK drawing attention to inconsistencies in the EU regulatory framework, the EU will continue to ignore the real and present environmental risks associated with upgraded non-GM crops for biopolitical reasons. PMID- 17113666 TI - High density dental materials and radiotherapy planning: comparison of the dose predictions using superposition algorithm and fluence map Monte Carlo method with radiochromic film measurements. AB - BACKGROUND AND PURPOSE: During radiotherapy planning high density dental materials create a major challenge in determining correct dose distribution inside patients with head-and-neck tumors. PATIENTS AND METHODS: In this work we investigated the absorbed dose distribution inside a solid water slab phantom with embedded high density material irradiated by a 6MV photon beam of field size 10x10cm. We evaluated the absorbed dose distribution with three different techniques: superposition algorithm, radiochromic film, and the fluence map Monte Carlo (FMMC) method. RESULTS: The results obtained with radiochromic film and FMMC were in good agreement (within +/-5% of the dose) with one another. The superposition algorithm, which is often considered superior to other commercially available dose calculation algorithms, produced appreciably less accurate results than FMMC. In particular, downstream from the high density cerrobend inhomogeneity the superposition algorithm predicts a higher dose than the measurement does by at least 10-16% depending upon the size of the inhomogeneity and the distance from it. Upstream of the high density inhomogeneities the superposition algorithm predicts a lower than measured dose due to its failure to predict the dose enhancement close to the inhomogeneity interface. CONCLUSIONS: The delivered dose downstream from a high density inhomogeneity would be significantly less than the prescribed dose calculated by the superposition algorithm. The FMMC method which is based on a hybrid of the superposition algorithm input fluence data and Monte Carlo can be a useful tool in predicting dose in the presence of high density (e.g. dental) materials. PMID- 17113667 TI - Individual differences in chromosomal aberrations after in vitro irradiation of cells from healthy individuals, cancer and cancer susceptibility syndrome patients. AB - BACKGROUND: Radiosensitivity of normal tissue is a crucial factor of radiotherapy (RT)-related side effects. Here, we report the analysis of spontaneous and in vitro irradiation-induced chromosomal aberrations in 256,679 metaphases from 222 different individuals using three-color fluorescence in situ hybridization as a measure of radiosensitivity. MATERIALS AND METHODS: Samples were categorized into the following 6 groups: (1) healthy individuals, (2) cancer patients prior to radiotherapy, (3) RT-treated cancer patients, (4) individuals heterozygous or (5) homozygous for a mutation in the ataxia telangiectasia mutated (ATM) gene or in the Nijmegen breakage syndrome (NBS1) gene and (6) hypersensitive patients (outliers). RESULTS: A normal distribution of the number of chromosomal aberrations, measured as breaks per metaphase (B/m), was adopted for all examined groups. The mean value of the control group was 0.40B/m (SD+/-0.07). This value was lower compared to the mean breakage rate from 175 non-exposed (0.50+/ 0.12B/m) and pre-exposed (0.50+/-0.16B/m) cancer patients. Nineteen of the metaphase spreads from the analyzed cancer patients had a high number of chromosomal aberrations (1.04+/-0.29B/m) and were designated as a separate hypersensitive subgroup (outliers). The aberration frequency of this group was comparable to those of ATM or NBS1 heterozygotes (0.86+/-0.26B/m). The highest incidence of aberrations was observed in ATM and NBS1 homozygous patients (2.23+/ 1.03B/m). CONCLUSION: The frequency of break events in the analyzed groups resulted in a normal distribution with varying means and broadnesses defining a characteristic sensitivity pattern for each group. In the RT-relevant group of cancer patients, those patients who have cancer, about one-third of the normally distributed samples were determined to be sensitive as defined by the number of induced aberrations higher than the 99% confidence interval of the normal individual's Gaussian distribution. About 5% of these samples were outside of the 99% confidence interval for the RT-relevant group's normal distribution. These outliers with higher chromosomal breakage rates suggest a unique class of hypersensitive individuals that are susceptible to chromosomal damage and may be directly associated with an increased risk to suffer from radiotherapy-related complications. PMID- 17113668 TI - Comparison of helical, maximum intensity projection (MIP), and averaged intensity (AI) 4D CT imaging for stereotactic body radiation therapy (SBRT) planning in lung cancer. AB - BACKGROUND AND PURPOSE: To compare helical, MIP and AI 4D CT imaging, for the purpose of determining the best CT-based volume definition method for encompassing the mobile gross tumor volume (mGTV) within the planning target volume (PTV) for stereotactic body radiation therapy (SBRT) in stage I lung cancer. MATERIALS AND METHODS: Twenty patients with medically inoperable peripheral stage I lung cancer were planned for SBRT. Free-breathing helical and 4D image datasets were obtained for each patient. Two composite images, the MIP and AI, were automatically generated from the 4D image datasets. The mGTV contours were delineated for the MIP, AI and helical image datasets for each patient. The volume for each was calculated and compared using analysis of variance and the Wilcoxon rank test. A spatial analysis for comparing center of mass (COM) (i.e. isocenter) coordinates for each imaging method was also performed using multivariate analysis of variance. RESULTS: The MIP-defined mGTVs were significantly larger than both the helical- (p=0.001) and AI-defined mGTVs (p=0.012). A comparison of COM coordinates demonstrated no significant spatial difference in the x-, y-, and z-coordinates for each tumor as determined by helical, MIP, or AI imaging methods. CONCLUSIONS: In order to incorporate the extent of tumor motion from breathing during SBRT, MIP is superior to either helical or AI images for defining the mGTV. The spatial isocenter coordinates for each tumor were not altered significantly by the imaging methods. PMID- 17113669 TI - CT-guided intensity-modulated radiotherapy for bladder cancer: isocentre shifts, margins and their impact on target dose. AB - BACKGROUND AND PURPOSE: Image-guided radiotherapy (IGRT) has great potential to improve the treatment of tumour sites that exhibit large geometrical uncertainties such as the bladder. This study quantifies the size and direction of the daily 3D isocentre shift resulting from the use of IGRT and its impact on reducing the margins required. The changes to target dose coverage, delivered dose and dose homogeneity appearing when used in conjunction with intensity modulated radiotherapy (IMRT) are also assessed. MATERIALS AND METHODS: The study is performed using a series of 19 bladder cancer patients that underwent weekly repeat conventional CT scanning during their 6-7 weeks treatment course. The isotropic margin required to cover various percentages of the volumes of the repeat scans CTVs is found by growing the planning scan PTV by incrementing the margin in steps of 1mm until full coverage is obtained. The optimum daily isocentre shift is determined by moving the repeat scan CTV in 3D so that its volume that lies outside the planning CTV is minimised. Reduction in the isotropic margin after applying the optimum isocentre shift is found by repeating the above procedure. Individual optimum sizes of the six margins required in the sup/inf/ant/post/right/left directions are determined using a recently published empirical margin determination method. Finally, five-field IMRT plans are set up (that minimise the dose deviation throughout the target) using different planning CTV margin alternatives, the optimal isocentre shifts are applied and the target dose distributions are assessed after recalculation without changing any of the beam parameters. RESULTS: Without use of IGRT (i.e., without shifting to the optimal isocentre), an isotropic margin of 30mm is required to cover fully all the CTV volumes on the repeat scans for all patients. The direction of the optimum isocentre shift required is random with an average directional shift less than 1mm and is unlikely to exceed an absolute value of 15mm (average of 7mm). Applying the optimum isocentre shift reduces the isotropic margin to 16mm for full volume coverage. Determining the optimum margin individually in each of six orthogonal directions reduces the target volume by approximately 30% but requires complex daily planning. Applying the optimum isocentre shift to IMRT plans shows little change to the overall mean target dose of 100% (an average increase of 1%), but produces a spread in the daily mean dose ranging from 96% to 106%. The 3D dose variation over the target is within the 95-107% acceptable range in 72% of cases for a 12mm uniform margin, which increases to 91% if any deviation from the daily prescribed dose is removed. The minimum and maximum doses within the target can show significant changes. CONCLUSION: The use of IGRT in the treatment of bladder cancer leads to a marked reduction in the margins required. When used in conjunction with IMRT, the pre-treatment plan is shown to be acceptable for daily treatment (after shifting the isocentre) in terms of the resulting dose distribution provided the correct daily mean dose is delivered. That can be achieved by rescaling the daily monitor units for each treatment beam. PMID- 17113670 TI - Dose distribution in 3-dimensional conformal radiotherapy for prostate cancer: comparison of two treatment techniques (six coplanar fields and two dynamic arcs). AB - PURPOSE: To compare dose distribution for two techniques of 3-dimensional conformal radiotherapy (RT): 6-field technique (6F) and 2-dynamic arc therapy (2DA). METHODS AND MATERIALS: Thirty nonmetastatic prostate cancer patients were included. In each patient, two treatment plans were prepared: with six coplanar fields (45 degrees , 90 degrees , 135 degrees , 225 degrees , 270 degrees , 315 degrees ) and with two dynamic lateral 100 degrees -wide arcs (40-140 degrees , 220-320 degrees ). Dose-volume histograms (DVHs) were computed and mean area under curve (AUC) values were calculated for the DVHs of Planning Target Volume (PTV), rectum, urinary bladder and femoral heads. Doses given to 30% of rectum (DR(30)), to 60% of rectum (DR(60)), to 50% of bladder (DB(50)), to 50% of femoral head (DF(50)) and to 95% of PTV (DPTV(95)) were reported as a percentage of the total dose. RESULTS: Mean DR(30) and DR(60) for 6F and 2DA were 75.8%, 51.5% and 72.2%, 37.2%, respectively. Mean DB(50) for 6F and 2DA were 68% and 64.2%, respectively. Mean right DF(50) for 6F and 2DA were 35.4% and 45.5%, respectively. Mean DPTV(95) for 6F and 2DA were 99% and 99.2%, respectively. Mean AUCs of DVHs of rectum and urinary bladder were significantly higher for 6F (this was more evident for small PTV and in the intermediate dose range). Mean AUC of DVHs of PTV and femoral heads were significantly higher for 2DA. CONCLUSIONS: Both 6F and 2DA offer good dose distribution for PTV. 2DA allows for significantly better sparing of rectum and urinary bladder with slightly worse femoral head dose distribution. Further study is warranted in order to establish the clinical relevance of these differences. PMID- 17113671 TI - Comparison of dose-volume histograms of IMRT treatment plans for ethmoid sinus cancer computed by advanced treatment planning systems including Monte Carlo. AB - BACKGROUND AND PURPOSE: To recompute clinical intensity-modulated treatment plans for ethmoid sinus cancer and to compare quantitatively the dose-volume histograms (DVHs) of the planning target volume (PTV) and the optic organs at risk. MATERIAL AND METHODS: Ten step-and-shoot intensity-modulated treatment plans were enrolled in this study. Large natural and surgical air cavities challenged the calculation systems. Each optimized treatment plan was recalculated by two superposition convolution (TMS and Pinnacle) and a Monte Carlo system (MCDE). To compare the resulting DVHs, a one-way ANOVA for repeated measurements was performed and multiple pairwise comparisons were made. RESULTS: The tails of the PTV-DVHs were significantly higher for the Monte Carlo system. The DVHs of the critical organs displayed some statistically but not always clinically significant differences. For the individual patients, the three planning systems sometimes reproduced clinically discrepant DVHs that were not significantly different when averaged over all patients. CONCLUSIONS: Dose to air cavities contains computational uncertainty. As this dose is clinically irrelevant and optimizing it is meaningless, we recommended extracting the air from the PTV when constructing the PTV-DVH. The planning systems considered reproduce DVHs that are significantly different, especially in the tail region of PTV-DVHs. PMID- 17113672 TI - Motor brain regions are involved in the encoding of delayed intentions: a fMRI study. AB - In studies of prospective memory, recall of the content of delayed intentions is normally excellent, probably because they contain actions that have to be enacted at a later time. Action words encoded for later enactment are more accessible from memory than those encoded for later verbal report [Freeman, J.E., and Ellis, J.A. 2003a. The representation of delayed intentions: A prospective subject performed task? Journal of Experimental Psychology: Learning, Memory, and Cognition, 29, 976-992.]. As this higher assessibility is lost when the intended actions have to be enacted during encoding, or when a motor interference task is introduced concurrent to intention encoding, Freeman and Ellis suggested that the advantage of to-be-enacted actions is due to additional preparatory motor operations during encoding. Accordingly, in a fMRI study with 10 healthy young participants, we investigated whether motor brain regions are differentially activated during verbal encoding of actions for later enactment with the right hand in contrast to verbal encoding of actions for later verbal report. We included an additional condition of verbal encoding of abstract verbs for later verbal report to investigate whether the semantic motor information inherent in action verbs in contrast to abstract verbs activates motor brain regions different from those involved in the verbal encoding of actions for later enactment. Differential activation for the verbal encoding of to-be-enacted actions in contrast to to-be-reported actions was found in brain regions known to be involved in covert motor preparation for hand movements, i.e. the postcentral gyrus, the precuneus, the dorsal and ventral premotor cortex, the posterior middle temporal gyrus and the inferior parietal lobule. There was no overlap between these brain regions and those differentially activated during the verbal encoding of actions in contrast to abstract verbs for later verbal report. Consequently, the results of this fMRI study suggest the presence of preparatory motor operations during the encoding of delayed intentions requiring a future motor response, which cannot be attributed to semantic information inherent to action verbs. PMID- 17113673 TI - The role of noticing in prospective memory forgetting. AB - Two experiments used autonomic reactions (i.e., skin conductance responses; SCRs) in conjunction with behavioral responses to study retrieval processes in prospective memory. SCRs were recorded while participants performed a prospective memory task embedded in an ongoing task. Stimuli that received the same behavioral response (i.e., no prospective memory response) evoked different autonomic reactions as a function of whether they were versus were not prospective cues (Experiments 1 and 2) and as a function of whether they did versus did not share perceptual or conceptual features with prospective cues (Experiment 2). To the extent that SCRs provide an index of noticing a stimulus, increased SCRs for prospective cues and for stimuli that shared features with prospective cues (even though they were not responded to as prospective cues) provided evidence that noticing a stimulus is not invariably accompanied by recognizing the stimulus as a cue to perform an intended action. The results are consistent with the general 2-stages cue-focused view of prospective memory retrieval, which proposes that noticing a prospective cue prompts a directed memory search, which can result in recognizing the stimulus as a cue to perform an intended action and retrieving the intended action. PMID- 17113674 TI - Electrochemical biosensor for catechol using agarose-guar gum entrapped tyrosinase. AB - An electrochemical biosensor using tyrosinase was constructed for the determination of catechol. The enzyme was extracted from a plant source Amorphophallus companulatus and entrapped in agarose-guar gum composite biopolymer matrix. Catechol was determined by direct reduction of biocatalytically liberated quinone species at -0.1 V versus Ag/AgCl (3M KCl). The response was found to be linear and concentration dependent in the range of 6 x 10(-5) to 8 x 10(-4)M with a lower detection limit of 6 microM. It has reusability up to 20 cycles and a shelf life of more than 2 months when stored at 4 degrees C. PMID- 17113675 TI - Expression and self-assembly of cowpea chlorotic mottle virus-like particles in Pseudomonas fluorescens. AB - Coat protein of the cowpea chlorotic mottle virus (CCMV), a plant bromovirus, has been expressed in a soluble form in a prokaryote, Pseudomonas fluorescens, and assembled into virus-like particles (VLPs) in vivo that were structurally similar to the native CCMV particles derived from plants. The CCMV VLPs were purified by PEG precipitation followed by separation on a sucrose density gradient and analyzed by size exclusion chromatography, UV spectrometry, and transmission electron microscopy. DNA microarray experiments revealed that the VLPs encapsulated very large numbers of different host RNAs in a non-specific manner. The development of a P. fluorescens expression system now enables production of CCMV VLPs by bacterial fermentation for use in pharmaceutical or nanotechnology applications. PMID- 17113676 TI - Insulin glulisine imparts effective glycaemic control in patients with Type 2 diabetes. AB - INTRODUCTION: Insulin glulisine (glulisine) was evaluated versus regular human insulin (RHI) in Type 2 diabetes (T2DM) patients. METHODS: Patients previously on >6 months' continuous insulin treatment aged >or=18 years in a randomized, multinational, controlled, open-label, parallel group, 26-week study received twice-daily NPH insulin and either glulisine (0-15 min before breakfast and dinner; n=448) or RHI (30-45 min before breakfast and dinner; n=442) at least twice daily. RESULTS: Mean baseline characteristics were similar between groups. There were no differences in baseline to endpoint HbA(1c) reductions (glulisine: 0.32%; RHI: -0.35%; p=0.5726), and the non-inferiority of glulisine versus RHI was demonstrated (difference in adjusted mean change 0.03%; 95% CI: -0.07, 0.13). Postprandially, glulisine lowered plasma glucose significantly more versus RHI at 2h (14.14 mmol/L versus 15.28 mmol/L; p=0.0025) and excursions at 1h (3.99 versus 4.59; p=0.0151) and 2h (4.87 versus 6.03; p=0.0002). No between-group differences occurred in the frequencies and monthly rates of all symptomatic hypoglycaemia; nocturnal hypoglycaemia from Month 4 to treatment end was less frequent with glulisine versus RHI (9.1% versus 14.5%; p=0.029). CONCLUSION: Glulisine was non inferior to RHI in reducing HbA(1c) in T2DM. Glulisine demonstrated superior postprandial glucose control and was associated with fewer nocturnal hypoglycaemic episodes, indicating clinical benefits. PMID- 17113677 TI - Carbon nanotubes as nanomedicines: from toxicology to pharmacology. AB - Various biomedical applications of carbon nanotubes have been proposed in the last few years leading to the emergence of a new field in diagnostics and therapeutics. Most of these applications will involve the administration or implantation of carbon nanotubes and their matrices into patients. The toxicological and pharmacological profile of such carbon nanotube systems developed as nanomedicines will have to be determined prior to any clinical studies undertaken. This review brings together all the toxicological and pharmacological in vivo studies that have been carried out using carbon nanotubes, to offer the first summary of the state-of-the-art in the pharmaceutical development of carbon nanotubes on the road to becoming viable and effective nanomedicines. PMID- 17113678 TI - Urbanization and the ecology of wildlife diseases. AB - Urbanization is intensifying worldwide, with two-thirds of the human population expected to reside in cities within 30 years. The role of cities in human infectious disease is well established, but less is known about how urban landscapes influence wildlife-pathogen interactions. Here, we draw on recent advances in wildlife epidemiology to consider how environmental changes linked with urbanization can alter the biology of hosts, pathogens and vectors. Although urbanization reduces the abundance of many wildlife parasites, transmission can, in some cases, increase among urban-adapted hosts, with effects on rarer wildlife or those living beyond city limits. Continued rapid urbanization, together with risks posed by multi-host pathogens for humans and vulnerable wildlife populations, emphasize the need for future research on wildlife diseases in urban landscapes. PMID- 17113679 TI - Limits to evolution at range margins: when and why does adaptation fail? AB - What stops populations expanding into new territory beyond the edge of a range margin? Recent models addressing this problem have brought together population genetics and population ecology, and some have included interactions among species at range edges. Here, we review these models of adaptation at environmental or parapatric margins, and discuss the contrasting effects of migration in either swamping local adaptation, or supplying the genetic variation that is necessary for adaptation to continue. We illustrate how studying adaptation at range margins (both with and without hybridization) can provide insight into the genetic and ecological factors that limit evolution more generally, especially in response to current rates of environmental change. PMID- 17113680 TI - Modelling the long-term performance of zero-valent iron using a spatio-temporal approach for iron aging. AB - Zero-valent iron (ZVI) permeable reactive barriers (PRBs) have become popular for the degradation of chlorinated ethenes (CEs) in groundwater. However, a knowledge gap exists pertaining to the longevity of ZVI. The present investigation addresses this situation by suggesting a numerical simulation model that is intended to be used in conjunction with field or column tests in order to describe long-term ZVI performance at individual sites. As ZVI aging processes are not yet completely understood and are still subject to research, we propose a phenomenological modelling technique instead of a common process-based approach. We describe ZVI aging by parameters that characterise the extent and rate of ZVI reactivity change depending on the propagation of the precipitation front through ZVI. We approximate degradation of CEs by pseudo-first order kinetics accounting for the formation of partially dechlorinated products, and describe ZVI reactivity change by scaling the degradation rate constants. Three independent modelling studies were carried out to test the suitability of the conceptual and numerical model to describe the observations of accelerated column tests. All three tests indicated that ZVI reactivity declined with an increasing number of exchanged pore volumes. Measured and modelled concentrations showed good agreement, thereby proving that resolving spatial as well as temporal changes in ZVI reactivity is reasonable. PMID- 17113681 TI - Healthy eating: perceptions and practice (the ASH30 study). AB - Perceptions of healthy eating may influence food intake. Anthropometric and dietary data were collected from 197 respondents (average age 32.5 years: 2000/2001) in Northumberland (78%) and elsewhere in the UK (22%). A questionnaire and two 3-day food diaries were completed. Foods consumed were assigned to one of five food categories from The Balance of Good Health. This paper explores respondents' concepts of 'healthy eating' and responses to the statement, 'My eating patterns are healthy' and compares responses with measured intakes for each of the five food categories. Fifty-three respondents disagreed, 62 neither agreed nor disagreed and 82 agreed with the statement. Intakes of foods containing fat and/or sugar, fruit and vegetables and meat, fish and alternatives were significantly different between the three response groups. The 'agree' group had the highest intake of fruit and vegetables and the lowest intake of foods containing fat and/or sugar and meat, fish and alternatives. A significantly higher proportion of individuals from the highest socio-economic group agreed with the statement. Significantly more individuals with Body Mass Indexes in the two lower quartiles agreed with the statement. This paper shows a relationship between perceptions of eating patterns and socio-economic status, adiposity and measured food intake. PMID- 17113682 TI - A comprehensive framework for determining the cost of an emergency medical services system. AB - To determine the cost of an emergency medical services (EMS) system, researchers, policymakers, and EMS providers need a framework with which to identify the components of the system that must be included in any cost calculations. Such a framework will allow for cost comparisons across studies, communities, and interventions. The objective of this article is to present an EMS cost framework. This framework was developed by a consensus panel after analysis of existing peer reviewed and non-peer-reviewed resources, as well as independent expert input. The components of the framework include administrative overhead, bystander response, communications, equipment, human resources, information systems, medical oversight, physical plant, training, and vehicles. There is no hierarchical rank to these components; they are all necessary. Within each component, there are subcomponents that must be considered. This framework can be used to standardize the calculation of EMS system costs to a community. Standardizing the calculation of EMS cost will allow for comparisons of costs between studies, communities, and interventions. PMID- 17113683 TI - Tramadol/acetaminophen or hydrocodone/acetaminophen for the treatment of ankle sprain: a randomized, placebo-controlled trial. AB - STUDY OBJECTIVE: This randomized, multicenter study compares the analgesic efficacy and safety of tramadol/acetaminophen versus hydrocodone/acetaminophen versus placebo for the treatment of acute musculoskeletal pain caused by ankle sprain. METHODS: Adults were enrolled with ankle sprain with a diagnosis of partial ligament tear, pain visual analog scale score of 50 to 100 mm (0="no pain," 100="extreme pain"), and pain numeric rating scale score of 2 to 3 (0="none," 3="severe"). Patients reported pain intensity on these scales and pain relief (-1="pain worse," 4="complete relief") hourly for 4 hours after the first dose of tramadol/acetaminophen 75 mg/650 mg, hydrocodone/acetaminophen 7.5 mg/650 mg, or placebo, and daily for 5 days, with as-needed dosing. RESULTS: Tramadol/acetaminophen (n=192) and hydrocodone/acetaminophen (n=204) provided greater total pain relief than placebo (n=207; P<.001) during the first 4 hours (mean scores [95% confidence interval (CI)] 6.6 [95% CI 6.1 to 7.1], 6.8 [95% CI 6.3 to 7.3], and 5.4 [95% CI 4.9 to 5.9], respectively; possible range -4 to 16), decreased pain intensity during the first 4 hours, and increased average pain relief on days 1 to 5. No efficacy measure was significantly different between the tramadol/acetaminophen and hydrocodone/acetaminophen groups. Common adverse events included somnolence, nausea, dizziness, and vomiting. CONCLUSION: One or 2 capsules of 37.5 mg tramadol/325 mg acetaminophen and 1 capsule of 7.5 mg hydrocodone/650 mg acetaminophen were well tolerated, had comparable clinical utility, and were more effective than placebo in the management of acute musculoskeletal pain caused by ankle sprain. PMID- 17113684 TI - Implementing an HIV and sexually transmitted disease screening program in an emergency department. AB - STUDY OBJECTIVE: We assess the feasibility, effectiveness, and cost of routinely recommended HIV/sexually transmitted disease screening in an urban emergency department (ED). METHODS: From April 2003 to August 2004, patients aged 15 to 54 years were offered rapid HIV testing, and those aged 15 to 25 years were also offered gonorrhea and chlamydia testing (nucleic acid amplification), Monday through Friday, 11 am to 8 pm. Infected patients were referred for treatment and care. Prevalence, treatment rates, and cost were assessed. RESULTS: Among 3,030 patients offered HIV testing, 1,447 (47.8%) accepted, 8 (0.6%) tested positive, and 3 (37.5%) were linked to care. Among 791 patients offered sexually transmitted disease testing, 386 (48.8%) accepted, 320 provided urine (82.9%), 48 (15.0%) tested positive, and 42 (87.5%) were treated for gonorrhea or chlamydia. The program cost was $72,928. Costs per HIV-infected patient identified and linked to care were, respectively, $9,116 and $24,309; cost per sexually transmitted disease-infected patient treated was $1,736. The program cost for HIV/sexually transmitted disease screening was only $14,340 more than if we screened only for HIV. CONCLUSION: Through ED-based HIV/sexually transmitted disease screening, we identified and treated many sexually transmitted disease infected patients but identified few HIV-infected patients and linked even fewer to care. However, sexually transmitted disease screening can be added to HIV screening at a reasonable cost. PMID- 17113685 TI - B-vitamins reduce plasma levels of beta amyloid. AB - Elevated plasma homocysteine (tHcy) is a risk factor for Alzheimer's disease (AD), and thus B vitamins may have a role in the prevention of AD. The objective of this study was to determine if tHcy lowering vitamins decrease the circulating levels of A-beta protein 1-40 (A beta 40). We randomized 299 older men to treatment with 2mg of folate, plus 25mg of B6 and 400 microg of B12, or placebo. After 2 years of treatment the mean (S.E.) increase of A beta 40 was 7.0 pg/ml (8.4) in the vitamin group (4.9%), and 26.8 pg/ml (7.7) (18.5%) in the placebo group. We conclude that B vitamins may decrease the plasma level of A beta 40 and have a role in the prevention of AD. PMID- 17113686 TI - Mental health nursing students' relationships with the pharmaceutical industry. AB - INTRODUCTION: The medical profession's relationship with the pharmaceutical industry (PI) has come under increased scrutiny in recent years, however little is known on the subject in mental health nursing. AIMS: The study sought to investigate: (1) the frequency of contact between mental health nursing students and PI employees; (2) students' attitudes and beliefs about their relationship with the PI; (3) the range of 'gifts', promotional items and hospitality accepted or seen in clinical environments by students in a one year period; and (4) students' attitudes to 'gifts', promotional items and hospitality offered by the industry. METHOD: Employing a survey design, a 35-item questionnaire was distributed to 472 students at two universities in the UK. Data were analysed from 347 respondents by means of descriptive statistics and simple content analysis. RESULTS: The findings suggest that students have significant contact with the industry through one-to-one meetings with pharmaceutical representatives (PRs) and by attending events giving information on specific drugs or general mental health issues. Students also identified a number of benefits (e.g. receiving "up-to-date" information on new drugs) and problems (e.g. the potential influence exerted on practitioners to use their drugs) arising out of this contact. Most students (79.8%) had accepted some form of 'gift' from the industry but few (11.5%) believed it was unacceptable to do so. The presence of promotional items in the clinical environment was seen as advertising (84.4%) but few students (19.3%) believed clinical environment should be free of these items. Over half (57.1%) of the students believed that PRs did not always give unbiased information but thought that they and mental health nurses in general would be able to detect any bias. CONCLUSIONS: In parallel with medicine, the study has shown that the pharmaceutical industry has at least the potential to influence mental health nursing students. Within medicine this realisation has triggered a vigorous debate on how medical schools should respond to the promotional activities of the PI. We suggest this study goes some way to demonstrating there is a need for these issues to be debated in the education of mental health nurses. PMID- 17113687 TI - Suppression of diabetes in non-obese diabetic (NOD) mice by oral administration of a cholera toxin B subunit-insulin B chain fusion protein vaccine produced in silkworm. AB - Oral tolerance has been applied successfully as a potential therapeutic strategy for preventing and treating autoimmune diseases including type 1 diabetes. In this paper we constructed an edible vaccine consisting of a fusion protein composed of cholera toxin B subunit (CTB) and insulin B chain (InsB) that was produced in silkworm larvae. The silkworm larvae produced this fusion protein at levels of up to 0.97mg/ml of hemolymph as the pentameric CTB-InsB form, which retained the GM1-ganglioside binding affinity and the native antigenicity of CTB. Non-obese diabetic mice fed hemolymph containing microgram quantities of the CTB InsB fusion protein showed a prominent reduction in pancreatic islet inflammation and a delay in the development of diabetic symptoms. This study demonstrates that silkworm-produced CTB-InsB fusion protein can be used as an ideal oral protein vaccine for induction of immunological tolerance against autoimmune diabetes. PMID- 17113688 TI - High antibody prevalence in an unconventional ecosystem is related to circulation of a low-virulent strain of Japanese encephalitis virus. AB - Liu-Chiu islet, a relatively isolated ecosystem that is free of rice cultivation, has long considered free of Japanese encephalitis (JE). However, a new strain (T1P1) of JE virus was isolated from the mosquito, Armigeres subalbatus, and a rather broad distribution of neutralizing antibody has been reported on the islet, suggesting that the circulating virus could be an attenuated strain. In an assessment on 219 blood samples obtained from residents of Liu-Chiu islet, the positive rate of JEV-specific IgM antibodies decreased with age while that of neutralizing antibodies increased with age. Both antibodies were mainly responsive to the T1P1 strain since higher positive rates and titers of specific neutralizing antibodies are shown in this investigation. Importantly, the T1P1 strain is herein characterized as being broader in neutralizing virus strains, stable in genetic traits, and productive in Vero cells. Taken together, the JE virus strain endemically circulating on Liu-Chiu islet may have served as a natural form of a live-attenuated vaccine. As a result, it possibly can be utilized as a new and effective vaccine candidate in the future. PMID- 17113689 TI - Protective efficacy of recombinant OmpTS protein of Aeromonas hydrophila in Indian major carp. PMID- 17113690 TI - Manuka honey dressing: An effective treatment for chronic wound infections. AB - The battle against methicillin-resistant Staphylococcus aureus (MRSA) wound infection is becoming more difficult as drug resistance is widespread and the incidence of MRSA in the community increases. Manuka honey dressing has long been available as a non-antibiotic treatment in the management of chronic wound infections. We have been using honey-impregnated dressings successfully in our wound care clinic and on the maxillofacial ward for over a year. PMID- 17113691 TI - Conscious analgesia and sedation during orthognathic surgery: preliminary results of a method of preventing condylar displacement. AB - A patient who is unconscious and paralysed has a condyle that is different from what it would be were he awake and the same force applied. The occlusion may relapse as a result of changes in condylar position immediately after the removal of intermaxillary fixation (IMF). Examination of the occlusion and an understanding of the changes secondary to a condylar displacement can identify it reliably during the operation. A study group (n=78) and a control group (n=74) were chosen randomly from patients listed for bimaxillary orthognathic surgery. No local anaesthetic was infiltrated so as not to disturb proprioception. In the study group, the IMF was removed immediately after the fixation and the occlusions were checked with light digital pressure on the chin. The patients were then woken rapidly (maintaining the intubation) in a state of conscious analgesia and sedation and invited to open and close their mouths and to move the mandible laterally. If clinical examination of the passive and active movements of the mandible were suitable, the anaesthetics were topped up and the operation completed. "Conscious" analgesia and sedation is certainly a valid aid during orthognathic surgery. PMID- 17113692 TI - Rare presentation of Mycobacterium avium-intracellulare infection. AB - Mycobacterium avium-intracellulare causes clinical disease mainly in immunocompromised patients with long-standing pulmonary disease, the symptoms of which are identical to those of pulmonary tuberculosis. In patients with AIDS the infection is typically disseminated. Extrapulmonary infection by M. avium intracellulare is seen in children aged 1-5 years as cervicofacial lymphadenitis. Other extrapulmonary sites are less common and include musculoskeletal, maxillary sinus, mastoid, breast, small bowel, genitourinary tract and cornea. Such infection of the hard palate is rare. We present an unusual case of infection of the palate by M. avium-intracellulare in a 53-year-old diabetic man. To our knowledge, this infection presenting as a palatal lump in a patient without HIV or pre-existing lung disease has not been previously reported. PMID- 17113693 TI - Endoscopic treatment of maxillary sinus disease before grafting. AB - We present our experience of the treatment of four patients with maxillary sinus disease by endoscopic sinus surgery to restore the normal physiology of the sinus before grafting. PMID- 17113694 TI - Tuberculosis of parotid gland. PMID- 17113695 TI - Urban-rural differences in the socioeconomic deprivation--sexual behavior link in Kenya. AB - We compare the impact of socioeconomic deprivation on risky sexual outcomes in rural and urban Kenya. Quantitative data are drawn from the Demographic & Health Surveys (DHS) and qualitative data from the Sexual Networking and Associated Reproductive and Social Health Concerns study. Using two separate indicators of deprivation we show that, although poverty is significantly associated with the examined sexual outcomes in all settings, the urban poor are significantly more likely than their rural counterparts to have an early sexual debut and a greater incidence of multiple sexual partnerships. The disadvantage of the urban poor is accentuated for married women; those in Nairobi's slums are at least three times as likely to have multiple sexual partners as their rural counterparts. The implications of these findings are discussed. PMID- 17113696 TI - Vitamin A and beta-carotene inhibitory effect during 1,2-dimethylhydrazine induced hepatocarcinogenesis potentiated by 5-azacytidine. AB - 5-Azacytidine is being used for reactivation of tumor suppressor genes. However, its administration during DNA repair pontentiates hepatocarcinogenesis. We observed chemopreventive activities by vitamin A and beta-carotene during early hepatocarcinogenesis. Thus, in the present study we evaluated vitamin A and beta carotene chemopreventive potential during early hepatocarcinogenesis potentiated by 5-azacytidine. Wistar rats received vitamin A (VAA group), beta-carotene (betaCA group) or corn oil (CO and COA groups). After three weeks of treatment, all animals were initiated with 1,2-dimethylhydrazine. Twelve hours later VAA, betaCA and COA groups received a single dose of 5-Azc. Hepatocytes were selected/promoted by 2-acetylaminofluorene and 70% partial hepatectomy. All animals were sacrificed six weeks after initiation. Compared to CO group (without 5-azacytidine), COA group presented higher (p<0.05) nodule multiplicity, larger (p<0.05) gamma-GT positive lesions that occupied a larger (p<0.05) area of liver section. Compared to COA group, VAA group presented decreased (p<0.05) nodule multiplicity while betaCA group tended to present smaller gamma-GT positive lesions and to decrease occupied liver section. These results reinforce vitamin A and beta-carotene chemopreventive potential. Considering that 5-azacytidine potentiates hepatocarcinogenesis, more studies are needed to elucidate the efficacy and safety of this drug for cancer control. PMID- 17113697 TI - Chronic exposure of mice to environmentally relevant, low doses of cadmium leads to early renal damage, not predicted by blood or urine cadmium levels. AB - Mice were exposed to cadmium (Cd) concentrations ranging from 0 to 100mg CdCl(2)/l in the drinking water for 1, 4, 8, 16 and 23 weeks. Urine samples were taken regularly, Cd content was determined in blood, liver, kidney and urine and histological analyses of the kidney were performed. Kidney cortex Cd content increased linearly with time and dose, while blood levels reached a plateau at 8 weeks and liver at 16 weeks in mice exposed to 100mg CdCl(2)/l after which both started to decrease. Urinary Cd levels were not correlated with the kidney Cd content. A multivariate regression model taking into account the actual Cd intake, calculated from the volume of water taken in by each animal and the exposure concentration, confirmed that blood is an indicator of acute exposure, while kidney Cd content is a reliable indicator of chronic exposure. The urinary protein content was significantly increased from 16 weeks on in mice exposed to 100mg CdCl(2)/l (p<0.05), while other signs of proximal tubular damage (glucosuria, enzymuria) were not detected. Histologically more vacuoles and lysosomes were present in the proximal tubule cells with increasing time and dose. The results indicate that chronic exposure to low doses of Cd induced functional and histological signs of early damage at concentrations in or below the ones generally accepted as safe. Our study does not corroborate the statement that urine Cd levels are a reliable indicator of total Cd body burden, at least when the body burden is low. PMID- 17113698 TI - The role of acrylophosphonic acid monomers in the formation of hybrid layers based on self-etch adhesives. AB - The role of acrylophosphonic acid monomers in the formation of hybrid layers based on self-etch adhesives. OBJECTIVES: Our plan was to define the reaction products formed when an acrylophosphonic acid reacts with tooth hard tissue. Our aim was to describe the incorporation of the reaction products in the hybrid layer formed. METHODS: Potentiometric methods were used to measure acid dissociation constants and investigate calcium complex formation. Infrared spectroscopy and NMR were used to follow water contents and show transformation of phosphorous containing compounds. RESULTS: The acrylophosphonic acid contained in AdheSE (Ivoclar Vivadent, Schaan, Liechtenstein) is characterized by two acidities with pK(a1)=2.4 and pK(a2)=7.0, and interacts with calcium ions in a purely ionic fashion. When hydroxyapatite crystals are dissociated by the presence of AdheSE, brushite is formed together with a calcium cross-linked network of the phosphonate containing copolymer. CONCLUSION: The results give a new image of the hybrid layer where the adhesive behaves like an ionomer resin incorporating collagen but also minerals and salt bridges. PMID- 17113699 TI - Effect of operator skill in relation to microleakage of total-etch and self-etch bonding systems. AB - OBJECTIVES: To evaluate the influence of operator skill on microleakage in class V restorations using two-step bonding systems. METHODS: Two standardized box cavities were made on the buccal and lingual surfaces of 54 human bicuspid teeth, with the cervical margin in dentin. The teeth were randomly assigned to two groups according to the operator's skill. Students group: undergraduate students. Expert group: a dentist with 20 years of clinical experience in dental adhesion. The buccal cavities of each tooth were treated with Prime&Bond NT (two-step total etch system); the lingual cavities were treated with AdheSE (two-step self-etch system). All cavities were restored by a single calibrated operator with one bulk increment of resin composite (InTens; Ivoclar Vivadent). Specimens were thermocycled, immersed in 2% methylene blue and sectioned in a bucco-lingual plane in the middle of the restorations. They were then examined under a stereomicroscope and scored according to microleakage by two operators who were blind to the specimen preparation. The data was subjected to a multilevel statistical model. RESULTS: The microleakage resulting from the self-etch adhesive was similar in the student and in the expert groups. On the other hand, the total-etch adhesive microleakage within the expert group resulted lower than that within the student group. However, the interaction term skill x adhesive resulted statistically significant at the dentin margin (p=0.0474) but not at the enamel margin (p=0.1267). CONCLUSION: While the total-etch adhesive used in this study showed to be skill-sensitive, the self-etch one proved to be less skill sensitive in obtaining a reliable seal with dentin. PMID- 17113700 TI - Modulated photoactivation methods: Influence on contraction stress, degree of conversion and push-out bond strength of composite restoratives. AB - OBJECTIVES: Verify the influence of curing methods on contraction stress, stress rate, and degree of conversion (DC) of a restorative composite and on bond strength of composite restoratives. METHODS: For the stress test, composite (0.84 mm thick) was applied between two 5-mm diameter glass rods, mounted in a servohydraulic machine. Stress rate was taken by the value of stress/time at each second. DC was measured by micro-FTIR. Bond strength testing was performed using a push-out test. The C-factor in all tests was 3.0. Four curing methods were tested: continuous light (CL), soft-start (SS), and two pulse delay methods using different initial irradiances--150 mW/cm(2) (PD150) and 80 mW/cm(2) (PD80). Results were analyzed by ANOVA and Tukey's test (alpha=0.05). RESULTS: Stress values ranged from 7.9 MPa (PD80) to 10.3 MPa (CL). No statistical difference was verified among CL, SS, and PD150. PD80 presented statistically lower stress values compared to CL and SS. CL presented the highest maximum stress rate, followed by SS, PD150 and PD80. Mean DC values ranged from 54.2% (PD150) to 55.9% (PD80), with no difference observed among the methods. For the bond strength test, values ranged from 26.4 MPa (CL) to 35.5 MPa (PD150). PD150 and PD80 were both statistically superior to SS and CL. SS presented statistically higher bond strength compared to CL. CONCLUSIONS: Modulated curing methods were shown to be effective in reducing contraction stress rate and improving the strength of the bonded interface, and without compromising the DC of the restorative composite. PMID- 17113701 TI - Magnetoencephalography: in search of neural processes for visual motion information. AB - Magnetoencephalography (MEG) has become a standard approach to the investigation of human brain functions. This review starts with a brief review of the human visual system and studies on visual motion detection mechanisms is followed by the presentation of MEG studies that have contributed to the field. Emphasis is placed on the fact that because the neural activities measured in functional magnetic resonance imaging (fMRI) differ substantially from those measured in MEG -fMRI data cannot be used directly to estimate MEG signal sources. The basic ideas behind the methods of signal processing and analyses generally used in MEG studies are described and theoretical considerations of the neural mechanisms determining MEG response latency and amplitude changes are discussed. Here, scalar fields theory is proposed to explain MEG responses to incoherent motions, and the ways in which detection of complex motions such as transparency, rotation and expansion can be explained by this theory are also presented. Relationships between human behavioral reaction time and MEG response latency suggest a new concept underlying the reasons why humans are late in detecting slow motion. PMID- 17113702 TI - Prediction of protein mutant stability using classification and regression tool. AB - Prediction of protein stability upon amino acid substitutions is an important problem in molecular biology and the solving of which would help for designing stable mutants. In this work, we have analyzed the stability of protein mutants using two different datasets of 1396 and 2204 mutants obtained from ProTherm database, respectively for free energy change due to thermal (DeltaDeltaG) and denaturant denaturations (DeltaDeltaG(H(2)O)). We have used a set of 48 physical, chemical energetic and conformational properties of amino acid residues and computed the difference of amino acid properties for each mutant in both sets of data. These differences in amino acid properties have been related to protein stability (DeltaDeltaG and DeltaDeltaG(H(2)O)) and are used to train with classification and regression tool for predicting the stability of protein mutants. Further, we have tested the method with 4 fold, 5 fold and 10 fold cross validation procedures. We found that the physical properties, shape and flexibility are important determinants of protein stability. The classification of mutants based on secondary structure (helix, strand, turn and coil) and solvent accessibility (buried, partially buried, partially exposed and exposed) distinguished the stabilizing/destabilizing mutants at an average accuracy of 81% and 80%, respectively for DeltaDeltaG and DeltaDeltaG(H(2)O). The correlation between the experimental and predicted stability change is 0.61 for DeltaDeltaG and 0.44 for DeltaDeltaG(H(2)O). Further, the free energy change due to the replacement of amino acid residue has been predicted within an average error of 1.08 kcal/mol and 1.37 kcal/mol for thermal and chemical denaturation, respectively. The relative importance of secondary structure and solvent accessibility, and the influence of the dataset on prediction of protein mutant stability have been discussed. PMID- 17113703 TI - Advanced age is associated with poorer bladder cancer-specific survival in patients treated with radical cystectomy. AB - OBJECTIVE: Bladder cancer (BCa) is a disease of older persons, the incidence of which is expected to increase as the population ages. There is controversy, however, regarding the outcomes of radical cystectomy (RC), the gold standard treatment of high-risk BCa, in patients of advanced chronological age. The aim of our study was to assess the impact of patient age on pathological characteristics and recurrence-free and disease-specific survival following RC. METHODS: The records of 888 consecutive patients who underwent RC for transitional cell carcinoma (TCC) were reviewed. Age at RC was analyzed both as a continuous (yr) and categorical (< or =60 yr old, n=240; 60.1-70 yr old, n=331; 70.1-80 yr old, n=266; >80 yr old, n=51) variable. Logistic regression and survival analyses were performed. RESULTS: Higher age at RC, analyzed as a continuous or categorical variable, was associated with extravesical disease and pathological upstaging (all p<0.02). Older patients were less likely to receive postoperative chemotherapy (< or =60 yr: 32% vs. >80 yr: 14%, p=0.008). In both pre- and postoperative multivariate models, higher age at RC as a categorical variable was associated with BCa-specific survival (p<0.05). Patients >80 yr old had a significantly greater risk of disease recurrence than patients aged < or =60 yr (p<0.05). CONCLUSION: Greater patient age at the time of RC for BCa is independently associated with adverse outcomes. Better understanding of factors associated with postoperative outcomes in this growing segment of the population is necessary. Prospective corroboration and further refinement of similar analyses in other large datasets is needed. PMID- 17113704 TI - The complex structure of the smooth muscle layer of spermatic veins and its potential role in the development of varicocele testis. AB - OBJECTIVES: Varicocele, a dilatation of the pampiniform venous plexus, is considered to cause male infertility. The exact mechanism of varicocele development is not clarified yet. This study focused on the structure of varicocele veins, compared with normal spermatic veins, and its potential role in varicocele development. METHODS: Morphologic and immunohistochemical studies using antibodies against vWF and neurofilament-200 (NF-200) were performed on spermatic vein fragments of 20 varicocele patients and 40 normal spermatic cords. Casting preparation of veins was performed on five normal spermatic cords. RESULTS: Casting preparation frequently revealed circular constrictions of normal spermatic vein lumina. Histologic evaluation showed a strong longitudinal smooth muscle layer in the adventitia of large veins in addition to the circularly organised tunica media. Serial sections showed smooth muscle fibres branching from the outer longitudinal into the inner circular layer. Immunostaining for vWF revealed high vascularisation of this outer layer. Interestingly, the number of nerve fibres marked by NF-200 immunostaining was considerably higher in large veins compared to the testicular artery. The longitudinal smooth muscle layer was significantly degraded in the presence of varicocele grades I and II, and did not even exist in varicocele grade III. Correspondingly, the number of vasa vasorum and nerve fibres was reduced in varicocele veins. CONCLUSIONS: Our data show a complex smooth muscle organisation of spermatic veins, which serves the basis for a contractile mechanism, providing an effective blood transport through pampiniform plexus. This mechanism is obviously damaged in the varicocele. Molecular processes behind this impairment remain to be clarified. PMID- 17113705 TI - Modelling the dynamic structure of biological state-based systems. AB - The paper discusses the modelling aspects of systems with dynamic processes and dynamic structure. A combination of models bringing together the benefits of two paradigms, Population P Systems and Communicating X-machines, is introduced. A simple case study is used in order to illustrate the potential of the combined use of the two methods. PMID- 17113706 TI - Ca2+ but not H2O2 modulates GRE-element activation by the human mineralocorticoid receptor in HEK cells. AB - The mineralocorticcoid receptor (MR) plays an important role in salt and water homeostasis as well as during cardiovascular and renal fibrosis but little is known regarding its modulation by other signaling pathways. To investigate a possible modulation under controlled conditions we used human embryonic kidney (HEK) cells (devoid of endogenous MR) transfected with the human MR and measured transactivation with a GRE-SEAP-reporter construct. MR was compared to the glucocorticoid receptor (GR) as well as to MR lacking the N-terminal domains AB (MR(CDEF)). Chelation of cytosolic Ca2+ enhanced MR activity and SGK1-expression, whereas elevation of cytosolic Ca2+ with ionomycin or thapsigargin reduced MR activity. GR activity was not affected by ionomycin or thapsigargin. MR(CDEF) activity was not affected by chelation or elevation of cytosolic Ca2+. Inhibition of ERK1/2 activation by U0126 or activation of PKA by cAMP, previously shown to modulate MR and GR activity, did not affect MR(CDEF) activity either. H2O2<500micromol/l did not affect basal nor hormone-induced reporter activity. Higher concentrations exerted the same relative inhibitory effect on GRE-SEAP activity under basal conditions as in the presence of aldosterone-stimulated MR and elicited cytotoxic effects. Our data indicate that the genomic function of MR can be modulated by cytosolic Ca2+, PKA and ERK1/2 via an interaction with the AB domain. H2O2 seems not to affect relative MR activity directly under our experimental conditions. PMID- 17113707 TI - Growth inhibition and sensitization to cisplatin by zoledronic acid in osteosarcoma cells. AB - Since osteosarcoma is a drug-resistant disease, the aim of the present study was to explore the possible interest of therapeutic approaches including nitrogen containing biphosphonate zoledronic acid using osteosarcoma cell lines with different genetic backgrounds. Parental p53+/pRb+ U2-OS, p53-mutant U2-OS (U2 OS/175) and p53-/pRb- SAOS were sensitive to zoledronic acid with no significant differences in IC50 values. Analysis of cell cycle distribution revealed a time dependent shifting of U2-OS cells towards G2 phase with cell cycle arrest in G2 phase at 96 h of exposure to the compound. Conversely, U2-OS/175 and SAOS cells responded to treatment with transient cell accumulation in S phase up to 48-72 h, respectively. Cell lines were exposed to increasing concentrations of cisplatin alone or combined with sub-toxic doses of zoledronic acid. A growth inhibitory effect was seen after combined treatment in U2-OS, otherwise resistant to cisplatin up to 100 ng/ml. Zoledronic acid did not efficiently sensitized U2 OS/175 and SAOS to cisplatin, thereby suggesting that different behavior may depend on p53 mutation. This data was confirmed in U2-OS cells where p53 expression was downregulated by RNA interference. Present findings indicate occurrence of sensitization to cisplatin by zoledronic acid in wild-type p53 osteosarcoma cells but not in p53-null cells nor in cells expressing a dominant negative form of p53, supporting that wild-type p53 is required for synergistic interaction of cisplatin and zoledronic acid. PMID- 17113708 TI - Effects of concentration, head group, and structure of surfactants on the degradation of phenanthrene. AB - The effects of concentration, polar/ionic head group, and structure of surfactants on the biodegradation of polycyclic aromatic hydrocarbons (PAHs) in the aqueous phase, as well as their effects on the bacterial activity were investigated. The toxicity ranking of studied surfactants is: non-ionic surfactants (Tween 80, Brij30, 10LE and Brij35) 5%. The Ki67 immunoreactivity was below 25% in 53 tumors (62.4%) and above 25% in 32 (32.6%). Our results revealed the existence of a statistically significant relationship of DNA ploidy with nodal status (p = 0.042) and grade (p = 0.005). Adenocarcinomas and large cell carcinomas were more frequently encountered in x-ploid-multiploid tumors as compared to squamous cell carcinomas, which were more frequently peridiploid (p = 0.003). 5cER showed statistically significant association with nodal status (p = 0.037). Univariate analysis with respect to survival revealed significant association with stage (p < 0.001), nodal status (p < 0.001), tumor status (p < 0.001), DNA ploidy (p = 0.008) and 5cER (p = 0.0124). Multivariate analysis revealed stage and ploidy status as independent factors: peridiploid tumors were associated with better survival as compared to x-ploid-multiploid tumors (p = 0.022). CONCLUSION: Our results suggest that DNA ploidy, as determined by image analysis, provides an independent prognostic parameter for patients with NSCLC and thus, could be used to identify a subset of patients with more aggressive tumors. PMID- 17113722 TI - Is blinding the endoscopists to bowel preparations in randomized-controlled trials a reality? AB - BACKGROUND: A number of studies compared the quality, efficacy and tolerability of oral sodium phosphate (NaP) and polyethylene glycol (PEG)-based solutions in preparation for colonoscopy. The primary aim of this study was to explore whether endoscopists can be effectively blinded to the type of bowel preparation. METHODS: We recruited 3 experienced endoscopists and 57 outpatients (18-65 years old) undergoing colonoscopy. We randomized eligible patients to receive one of the two bowel preparations. Endoscopists who performed the tests were blinded to the type of preparation, and made their best judgment on the type and quality of the bowel preparation. RESULTS: Forty-five patients completed the study. The overall correct estimation of the type of bowel preparation was 60.0% (95% CI; 45.5%, 73.0%). The cleansing quality did not differ between the two preparations. Patients found oral NaP solution much easier to take (81.8% versus 36.4%; P = 0.005) and the PEG-based group tended to have more nausea or vomiting. 47.6% of patients in the PEG group indicated they would prefer to try another bowel preparation in the future compared to 4.5% in the oral NaP group (P = 0.002). We stopped the study after an interim analysis indicating that more than 600 patients would be required to detect statistically significant differences in the primary aim. CONCLUSION: Our findings suggest that blinding of endoscopists in clinical trials comparing oral NaP to PEG had a relatively low likelihood of bias. The study also suggests that oral NaP is easier to take and more tolerable than PEG without impairing cleansing quality. PMID- 17113723 TI - Characterization of human breast epithelial cells by confocal Raman microspectroscopy. AB - BACKGROUND: The past decade has seen an explosion of interest in utilizing Raman spectroscopy in cancer diagnosis, due to its capability to probe changes in the biochemical composition of tissue that accompany disease progression. However, most of the existing methods used multivariate statistical analysis/chemometrics to differentiate normal and diseased tissues, which did not identify the compositional and chemical changes associated with the tumorigenic transition explicitly; also the sub-cellular level spatial resolution achievable through Confocal Raman microscopy was not fully utilized. METHODS: Confocal Raman microspectroscopy was used to characterize normal and transformed human breast epithelial cell lines. Key molecular components (DNA, RNA, and proteins) were extracted from cell nuclei and their Raman spectra were measured and used as a basis set to fit the spectra of cell nuclei. Contributions of each component and their relative contents were evaluated based on the fitting coefficients. RESULTS: Spectrum-fitting revealed that DNA duplication activities in tumorigenic cell nuclei are significantly higher than in normal cells. The fitting coefficients could serve as good spectral markers for disease state identification. CONCLUSIONS: A spectroscopic approach that yields compositional information of cell nuclei could be a powerful tool for rapid cell characterization and assessment of cellular activities at the sub-cellular level. PMID- 17113724 TI - Large genomic BRCA2 rearrangements and male breast cancer. AB - BACKGROUND: Germ-line mutations of the BRCA2 gene are the highest known risk factors for male breast cancer (MBC). Mutations in BRCA2 are mainly point mutations in contrast to BRCA1 in which large genomic rearrangements are quite common. In recent literature, however, genomic alterations of BRCA2 have been linked especially to male breast cancer families. We wanted to screen large genomic deletions and duplications of BRCA2 among Finnish male breast cancer patients. METHODS: We used multiplex ligation-dependent probe amplification (MLPA) to detect large genomic rearrangements in the BRCA2 gene among 36 unselected Finnish male breast cancer patients previously tested and found negative for Finnish BRCA1 and BRCA2 founder mutations. RESULTS: No genomic mutations of BRCA2 nor CHEK2*1100delC point mutations, also included in the assay, were found in this study. CONCLUSION: Large genomic BRCA2 rearrangements were not found among our 36 Finnish male breast cancer patients. Screening of large BRCA2 rearrangements is not likely to be advantageous in Finland. PMID- 17113725 TI - Evidence for an association of TP53 codon 72 polymorphism with breast cancer risk. AB - BACKGROUND: A common Arg/Pro polymorphism at codon 72 of the TP53 gene has been investigated as a risk factor for cancer in different populations. So far, the results have been controversial. Our purpose was to investigate the association of this polymorphism with breast carcinoma in women from Southern Brazil, a high risk area for breast cancer. METHODS: Blood samples collected from 118 women with primary breast carcinoma and from 202 female blood donors were analyzed through polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing. RESULTS: The relative frequency of each allele was 0.75 for Arg and 0.25 for Pro in patients with cancer, and 0.62 for Arg and 0.38 for Pro in normal controls (P < 0.001). The Arg/Arg genotype was significantly associated with an increased risk for breast cancer (OR 2.9; 95% CI 1.43-3.6; P < 0.002). No correlation between the genotype distribution and specific prognostic predictors for the disease outcome was observed. DISCUSSION: TP53 codon 72 polymorphism might be implicated in breast carcinogenesis, with the Arg/Arg genotype being associated with an increased susceptibility for this malignancy. PMID- 17113726 TI - Analgesic activity of Calotropis gigantea flower. AB - The alcoholic extract of the flowers of Calotropis gigantea was administered orally and explored for its analgesic activity in chemical and thermal models in mice. In acetic acid induced writhing test, an inhibition of 20.97% and 43.0% in the number of writhes was observed at the doses of 250 and 500 mg/kg, respectively. In the hot plate method the paw licking time was delayed. The analgesic effect was observed after 30 min of dose administration which reached its maximum after 90 min. PMID- 17113727 TI - Multiple roles for MSH2 in the repair of a deletion mutation directed by modified single-stranded oligonucleotides. AB - The mechanism by which modified single-stranded oligonucleotides (MSSOs) direct base changes in genes is not completely understood, but there is evidence that DNA damage, repair and cell cycle checkpoint proteins are involved in the targeted nucleotide exchange (TNE) process. We are interested in the role of the mismatch repair protein, Msh2 in the correction of a frameshift mutation in both yeast and mammalian cells. We show that this protein exerts different and opposing influences on the TNE reaction in MSH2 deficient yeast compared to MSH2( /-) mammalian cells and in wild-type cells that have RNAi silenced Msh2. Data from yeast show a 10-fold decrease in the targeting frequency whereas mammalian cells have an elevated correction frequency. These results show that in yeast this protein is required for efficient targeting and may play a role in mismatch recognition and repair. In mammalian cells, Msh2 plays a suppressive role in TNE reaction by either precluding the oligonucleotide annealing to the target gene or by maintenance of a cell cycle checkpoint induced by the MSSO itself. These results reveal that the mechanism of TNE between yeast and mammalian cells is not conserved, and demonstrate that the suppression of the TNE reaction can be bypassed using RNAi against MSH2 designed to knockdown its expression. PMID- 17113728 TI - Development of an improved species specific PCR test for detection of Haemophilus parasuis. AB - A PCR test for identification of Haemophilus parasuis was optimized using the 16S rDNA sequences of the 15 serotype reference strains of H. parasuis. The test was evaluated on a collection of 218 Danish field isolates as well as on 81 representatives of 27 other species, including genetically affiliated species within Pasteurellaceae. In addition, DNA preparations from 56 H. parasuis isolates from North America were included. To obtain a test that was specific for H. parasuis, a multiplex PCR using 3 different primers was developed. The PCR test produced an amplicon of approximately 1090 bp only with representatives of H. parasuis. The test was further evaluated on 55 clinical samples from 16 Danish pigs suspected for being infected with H. parasuis, showing polyserositis or septicemia at autopsy as well as on 492 nasal swabs. The test was compared with the performance of a PCR test earlier published by Oliveira et al. [Oliveira, S., Galina, L., Pijoan, C., 2001. Development of a PCR test to diagnose Haemophilus parasuis infections. J. Vet. Diagn. Invest. 13, 495-501]. The sensitivity of the present PCR test was found to be slightly lower when applied on clinical samples from diseased pigs and 10-fold lower when tested on pure cultures of H. parasuis (5CFU and 0.5CFU/PCR reaction, respectively). Addition of 1.4 x 10(5) Escherichia coli to each PCR tube did not alter the sensitivity of the tests. No difference in sensitivity of the tests was observed when tested on purified DNA. On the other hand, the present PCR test was found to be 100% species specific for H. parasuis, in contrast to the PCR test of Oliveira et al., which also tested positive for strains belonging to A. indolicus, A. porcinus, and A. minor, species commonly occurring in the upper respiratory tract. However, when the PCR test of Oliveira et al. is used on samples from systemic locations the chances for false positive results are apparently low. The present PCR test represents a rapid and reliable method for genetically based identification of H. parasuis. The high species specificity of the test makes it suitable for detection of H. parasuis in clinical samples, regardless of the presence of affiliated species and contaminating flora. As the two PCR tests differ in sensitivity and specificity, the use of both PCR tests for different purposes is a possibility. PMID- 17113729 TI - Genetic variation of foot-and-mouth disease virus isolates recovered from persistently infected water buffalo (Bubalus bubalis). AB - Genetic variation of foot-and-mouth disease virus (FMDV) isolates, serotype O, recovered serially over a 1-year period from persistently infected buffalos was assessed. The persistent state was established experimentally with plaque purified FMDV, strain O(1)Campos, in five buffalos (Bubalus bubalis). Viral isolates collected from esophageal-pharyngeal (EP) fluids for up to 71 weeks after infection were analyzed at different times by nucleotide sequencing and T(1) RNase oligonucleotide fingerprinting to assess variability in the VP1-coding region and in the complete genome, respectively. Genetic variation increased, although irregularly, with time after infection. The highest values observed for the VP1-coding region and for the whole genome were 2.5% and 1.8%, respectively. High rates of fixation of mutations were observed using both methodologies, reaching values of 0.65 substitutions per nucleotide per year (s/nt/y) and 0.44s/nt/y for nucleotide sequencing and oligonucleotide fingerprinting, respectively, when selected samples recovered at close time periods were analyzed. The data herein indicate that complex mixtures of genotypes may arise during FMDV type O persistent infection in water buffalos, which can act as viral reservoirs and also represent a potential source of viral variants. These results fit within the quasi-species dynamics described for FMDV, in which viral populations are constituted by related, non-identical genomes that evolve independently from each other, and may predominate at a given time. PMID- 17113730 TI - Influence of intensive and extensive breeding on lactic acid bacteria isolated from Gallus gallus domesticus ceca. AB - In the present study, lactic acid bacteria (LAB) from the cecum of chickens bred either under intensive (commercial broilers) or extensive (free-range) conditions were isolated, identified and some of their probiotic characteristics determined. The LAB identified by 16S-23S rRNA PCR-ARDRA were mainly of Lactobacillus species and to a lesser extent of Enterococcus spp. for all animals. Free-range chickens showed a higher presence of Lactobacillus acidophilus while Lactobacillus reuteri and Lactobacillus johnsonii were more frequently recovered from commercial broilers. Lactobacillus crispatus was found only in commercial broilers, Lactobacillus vaginalis and Lactobacillus agilis only in free-range chickens and Lactobacillus salivarius in both types. Enterococcus isolates from ceca of commercial broilers showed a higher resistance to antimicrobial drugs. Lactobacillus isolates from free-range chickens presented a higher frequency of in vitro antagonistic activity against selected pathogens than from commercial broilers. All LAB isolates had predominantly non-hydrophobic surfaces, but with variations depending on age of the chickens and breeding conditions. Animal breeding caused variation on composition, antimicrobial susceptibility, antagonistic activity and surface hydrophobicity of LAB from chicken cecum. LAB isolates from ceca of free-range chickens have potential as probiotic agents, which may be used in the future as replacing the use of antimicrobials as growth promoters. PMID- 17113731 TI - Comparison of heart rate responses during cortical and subcortical arousals in term and preterm infants. AB - The aim of this study was to determine whether prematurity affects heart rate responses during spontaneous arousals. Polygraphic recordings were performed during undisturbed daytime naps in 35 preterm infants (gestational age at birth 32+/-2 weeks) and 35 term infants. Arousals were scored according to the recommendations of the International Paediatric Work Group on Arousals and categorized either as cortical arousals (CA) or subcortical arousals (SCA). Heart rate (HR) and respiratory frequency (RF) were measured during arousal and during the 10-s and 30-s period before and after arousal. Changes in HR and RF were expressed as the percentage of modification normalized for the 30-s period preceding arousal. Altogether, 122 arousals in preterm infants (66 CA, 56 SCA) and 105 arousals in term infants (57 CA, 48 SCA) were scored. Mean duration of the arousal period was 9+/-4 s and 8+/-3 s, respectively. In term infants, a significant increase in HR during arousal could be shown (11.3+/-8.2%; p<0.001), whereas this increase was significantly greater during CA compared to SCA (13.7+/ 6.2% versus 8.4+/-9.4%; p<0.001). In contrast, HR decreased during arousal in preterm neonates (-3.9+/-19.3%; p<0.05). These findings suggest that cardiovascular control seems to be maturationally delayed in preterm infants, which may contribute to their increased risk for Sudden Infant Death Syndrome (SIDS). PMID- 17113732 TI - Alginate microspheres of isoniazid for oral sustained drug delivery. AB - In the present study, spherical microspheres able to prolong the release of INH were produced by a modified emulsification method, using sodium alginate as the hydrophilic carrier. The shape and surface characteristics were determined by scanning electron microscopy using gold sputter technique. Particle sizes of both placebo and drug-loaded formulations were measured by SEM and the particle size distribution was determined by an optical microscope. The physical state of the drug in the formulation was determined by differential scanning calorimetry (DSC). The release profiles of INH from microspheres were examined in simulated gastric fluid (SGF pH 1.2) and simulated intestinal fluid (SIF pH 7.4). Gamma scintigraphic studies were carried out to determine the location of microspheres on oral administration and the extent of transit through the gastrointestinal tract (GIT). The microspheres had a smoother surface and were found to be discreet and spherical in shape. The particles were heterogeneous with the maximum particles of an average size of 3.719mum. Results indicated that the mean particle size of the microspheres increased with an increase in the concentration of polymer and the cross-linker as well as the cross-linking time. The entrapment efficiency was found to be in the range of 40-91%. Concentration of the cross linker up to 7.5% caused increase in the entrapment efficiency and the extent of drug release. Optimized isoniazid-alginate microspheres were found to possess good bioadhesion (72.25+/-1.015%). The bioadhesive property of the particles resulted in prolonged retention in the small intestine. Microspheres could be observed in the intestinal lumen at 4h and were detectable in the intestine 24h post-oral administration, although the percent radioactivity had significantly decreased (t(1/2) of (99m)Tc=4-5h). Increased drug loading (91%) was observed for the optimized formulation suggesting the efficiency of the method. Nearly 26% of INH was released in SGF pH 1.2 in 6h and 71.25% in SIF pH 7.4 in 30h. No significant drug-polymer interactions were observed in FT-IR studies. Dissolution and gamma-scintigraphy studies have shown promising results proving the utility of the formulation for enteric drug delivery. PMID- 17113733 TI - Influence of carrier on the performance of dry powder inhalers. AB - The aim of this work is to study carriers which can become alternatives to monohydrate lactose in dry powder inhalers and to consider particle parameters that influence adhesion between drug and carrier in dry powder inhalers. Different forms of mannitol, lactose and maltitol were mixed with either terbutaline sulphate or formoterol fumarate. The blends were submitted to different adhesion tests where drug detachment from the carrier was obtained either through mechanical vibration or by aspiration. Parameters like particle shape, roughness, amorphous content and cristalline form may affect interactions between drug and carrier. In our case, crystallized forms of the carrier offered lower adhesion but better release of the active ingredient than spray-dried forms. The crystallized mannitol produced maximal fine particle dose. The blends of the mannitols and the two active ingredients gave different results. The two techniques used to assess the adhesion of drugs to carrier particles provide complementary information about drug/carrier interactions and detachment. The mechanical sieving allows to assess blend stability and the air-jet sieving makes it possible to determine how easily the drug separates from carrier. For the drugs tested, the results of fine particle doses are in agreement with the Alpine air-jet sieve results. The tests used are helpful for the choice of a new carrier in the field of the development of new carriers for dry powder inhalers. PMID- 17113734 TI - Synthesis and characterization of xylan-coated magnetite microparticles. AB - This work evaluates an experimental set-up to coat superparamagnetic particles in order to protect them from gastric dissolution. First, magnetic particles were produced by coprecipitation of iron salts in alkaline medium. Afterwards, an emulsification/cross-linking reaction was carried out in order to produce magnetic polymeric particles. The sample characterization was performed by X-ray powder diffraction, laser scattering particle size analysis, optical microscopy, thermogravimetric analysis and vibrating sample magnetometry. In vitro dissolution tests at gastric pH were evaluated for both magnetic particles and magnetic polymeric particles. The characterization data have demonstrated the feasibility of the presented method to coat, and protect magnetite particles from gastric dissolution. Such systems may be very promising for oral administration. PMID- 17113735 TI - Diuretic activity of the aqueous extracts of Carum carvi and Tanacetum vulgare in normal rats. AB - In the Moroccan traditional medicine, the ripe fruits of Carum carvi L. (Apiaceae) and the leaves of Tanacetum vulgare L. (Asteraceae/Compositae), two widely available plant materials, are used as diuretics. Since, the diuretic activity of these substances has not been investigated in scientifically controlled studies, the aim of the present study was to evaluate the diuretic potential of aqueous extracts of Carum carvi fruit (caraway) and the leaves of Tanacetum vulgare (tansy) in normal rats after acute and sub-chronic oral administration. Water extracts of Carum carvi and Tanacetum vulgare (100 mg/kg) or the reference drug, furosemide (10 mg/kg) were administrated orally to male Wistar rats and their urine output was quantitated at several intervals of time after the dose. After single doses of the extracts of both caraway seeds and tansy leaves, urine output was significantly increased at all time points, and at 24 h after the dose, the total volume of urine excreted was similar for the plant extracts and furosemide. Both extracts increased urinary levels of Na(+) and K(+), to about the same extent, while furosemide increased urinary levels of only Na(+) and decreased urinary K(+). Despite changes in urinary excretion of the electrolytes, plasma Na(+) and K(+) levels were not affected by any of the three substances. In the 8-day sub-chronic study, all three substances induced significant diuresis and natriuresis; only tansy increased urinary potassium excretion. The plant extracts did not appear to have renal toxicity or any other adverse effects during the study period. In conclusion, water extracts of both Carum carvi and Tanacetum vulgare have strong diuretic action confirming their ethnopharmacological use. From the pattern of excretion of water, sodium and potassium, it may be deduced that there are atleast two types of active principals present in these extracts, one having a furosemide-like activity and the other a thiazide-like activity. PMID- 17113736 TI - Analgesic activity of affinin, an alkamide from Heliopsis longipes (Compositae). AB - Heliopsis longipes (Compositae) is a Mexican plant used as analgesic in pain toothache. A solution of 10mug/ml of dichloromethane extract from this plant showed analgesic activity determined by means of GABA release in mice brain slices. Through a bioassay-directed separation, fractions G-1, G-2, G-4 and G-6 at the same concentration were active. Affinin was the unique and common active compound, and evoke the GABA release 0.5min after administration at 1x10(-4)M concentration. Inactive compound were undeca-2E-en-8,10-dyinoic acid isobutylamide, hinokinin, 2'-hydroxyhinokinin, 3beta-sn-glyceroyl-(1'' palmitoxy)urs-12-ene, 13(18)-ursen-3beta-ol, 13(18)-ursen-3beta-acetate, beta sitosterol and stigmasterol. The analgesic activity of Heliopsis longipes could be associated to affinin. PMID- 17113737 TI - Antimetastatic activities of Selaginella tamariscina (Beauv.) on lung cancer cells in vitro and in vivo. AB - Selaginella tamariscina is a traditional Chinese herb for the therapy of chronic trachitis and has been approved some anti-tumor activity. However, the anti metastasis effects of Selaginella tamariscina in the lung cancer have not been understood clearly. The objectives of study were to investigate the effects of the Selaginella tamariscina extracts (STE) on the invasion and motility of highly metastatic A549 and Lewis lung carcinoma (LLC) cells. To further investigate the precise involvement of STE in tumor metastasis, A549 and LLC cells were treated with STE at various concentrations (0-100 microg/mL) for a specified period. The results from zymography showed that a STE treatment decreased (p<0.05) the expressions of matrix metalloproteinase (MMP)-2, -9 and urokinase plasminogen activator (u-PA) in a dose-dependent manner in the A549 and LLC cell. Meanwhile, their endogenous inhibitors, which are tissue inhibitor of metalloproteinase-2 (TIMP-2) and plasminogen activator inhibitor-1 (PAI-1), were increased in the A549 cell. Furthermore, the inhibitory effect of STE on the growth and metastasis of LLC cells in vivo was also proven. These results demonstrated that STE could be a candidate antimetastatic agent against lung cancer. PMID- 17113738 TI - In vitro and in vivo anthelmintic activity of crude extracts of Coriandrum sativum against Haemonchus contortus. AB - In vitro anthelmintic activities of crude aqueous and hydro-alcoholic extracts of the seeds of Coriandrum sativum (Apiaceae) were investigated on the egg and adult nematode parasite Haemonchus contortus. The aqueous extract of Coriandrum sativum was also investigated for in vivo anthelmintic activity in sheep infected with Haemonchus contortus. Both extract types of Coriandrum sativum inhibited hatching of eggs completely at a concentration less than 0.5 mg/ml. ED(50) of aqueous extract of Coriandrum sativum was 0.12 mg/ml while that of hydro-alcoholic extract was 0.18 mg/ml. There was no statistically significant difference between aqueous and hydro-alcoholic extracts (p>0.05). The hydro-alcoholic extract showed better in vitro activity against adult parasites than the aqueous one. For the in vivo study, 24 sheep artificially infected with Haemonchus contortus were randomly divided into four groups of six animals each. The first two groups were treated with crude aqueous extract of Coriandrum sativum at 0.45 and 0.9 g/kg dose levels, the third group with albendazole at 3.8 mg/kg and the last group was left untreated. Efficacy was tested by faecal egg count reduction (FECR) and total worm count reduction (TWCR). On day 2 post treatment, significant FECR was detected in groups treated with higher dose of Coriandrum sativum (p<0.05) and albendazole (p<0.001). On days 7 and 14 post treatment, significant FECR was not detected for both doses of Coriandrum sativum (p>0.05). Significant (p<0.05) TWCR was detected only for higher dose of Coriandrum sativum compared to the untreated group. Reduction in male worms was higher than female worms. Treatment with both doses of Coriandrum sativum did not help the animals improve or maintain their PCV while those treated with albendazole showed significant increase in PCV (p<0.05). PMID- 17113739 TI - The in vitro activity of geraniin and 1,3,4,6-tetra-O-galloyl-beta-D-glucose isolated from Phyllanthus urinaria against herpes simplex virus type 1 and type 2 infection. AB - Phyllanthus urinaria Linnea (Euphorbiaceae) is a widely used traditional medicinal plant by oriental countries and has been reported to possess various biological activities. Previously, the acetone extract from Phyllanthus urinaria was found to inhibit herpes simplex virus (HSV) infection. In this study, geraniin and 1,3,4,6-tetra-O-galloyl-beta-D-glucose (1346TOGDG), both of which were isolated from the acetone extract of Phyllanthus urinaria, were examined for their activity against HSV-1 and HSV-2 in vitro. Results showed that geraniin actively suppressed HSV-2 infection, whereas 1346TOGDG effectively inhibited HSV 1 infection. The 50% inhibitory concentration (IC(50)) was 18.4+/-2.0 microM for geraniin against HSV-2 infection, and 19.2+/-4.0 microM for 1346TOGDG against HSV 1. No toxic effect towards the host cell was observed at the antiviral concentrations. In conclusion, geraniin and 1346TOGDG were found to inhibit HSV-1 and HSV-2 multiplication at different magnitudes of potency. PMID- 17113740 TI - Immunomodulatory activity of the rhizomes of Impatiens pritzellii var. hupehensis on collagen-induced arthritis mice. AB - Impatiens pritzellii Hook. f. var. hupehensis Hook. f. (Balsaminaceae) has been well-known and widely used in China as an anti-rheumatoid arthritis (anti-RA) herb. In this present study, mice with collagen-induced arthritis (CIA) have been treated with the methanol (MeOH) extract (0.56, 1.12, 1.68 and 2.24 g/kg body weight) and the n-butanol (BuOH) fraction (0.13, 0.27, 0.40 and 0.53 g/kg body weight) of the rhizomes of Impatiens pritzellii orally for 3 weeks. The progression of CIA was evaluated by macroscopic scoring. Administration of the MeOH extract at dose of 1.12 g/kg and the BuOH fraction at 0.53 g/kg suppressed the development of CIA in mice significantly. The spleen and thymus indexes were measured and the levels of IgG, IL-10, INF-gamma and IL-18 in the serum of CIA mice were examined after the treatment of the MeOH extract (1.12 and 1.68 g/kg body weight) and the BuOH fraction (0.40 and 0.53 g/kg body weight). Administration of the MeOH extract and the BuOH fraction of Impatiens pritzellii decreased the spleen and thymus indexes, down-regulated the levels of IgG, INF gamma, IL-18, and up-regulated the concentration of IL-10 in the serum of mice with CIA. From the results, it was concluded that administration of Impatiens pritzellii had obviously therapeutic effects on RA including immunomodulatory activity. Moreover, the BuOH fraction exerted the activity of anti-RA of Impatiens pritzellii. PMID- 17113741 TI - In vivo study of propolis supplementation effects on antioxidative status and red blood cells. AB - In vivo study has been conducted on 47 healthy women and men in order to investigate whether daily intake of powdered propolis extract during 30 days has any influence on the following blood parameters: activity of superoxide dismutase, glutathione peroxidase and catalase, concentration of plasma malondialdehyde, total cholesterol, low- and high-density lipoprotein cholesterol, triglycerides, glucose, uric acid, ferritin and transferrin, together with routine red blood cell parameters. The effect of daily propolis intake seems to be time and gender related. For the men test group after the initial 15 days of propolis treatment, 23.2% (p=0.005) decrease in concentration of malondialdehyde was observed. After 30 days of treatment, statistically significant (p=0.010) 20.9% increase in superoxide dismutase activity and change in some of the red blood cell parameters were detected. For the women test group, the propolis treatment did not induce a change in any of the measured parameters. PMID- 17113742 TI - Distortion product otoacoustic emissions during the first year in term infants: a longitudinal study. AB - Understanding of any age-related differences in distortion product otoacoustic emissions (DPOAEs) during infancy is important for the use of DPOAEs in detection of cochlear impairment in infants. We studied DPOAEs at 10 frequencies of f2 primary tone between 500 Hz to 10 kHz longitudinally during the first year of life in 70 ears of 35 normal term infants. On days 3-5 after birth DPOAE pass rates ranged between the highest 98.6% at f2 frequencies 6-10 kHz and the lowest 22.9% at 750 Hz. The higher the frequency, the higher was the pass rate. At 6 months the pass rates at various frequencies were generally similar to those on days 3-5, and were greater than 91% across the frequencies of 3-10 kHz. At 12 months the pass rates were 100% or near 100% across 3-10 kHz. During the first year the pass rate was always very low at 750 Hz (<40%) and tended to decrease at 1 kHz with the increase in age. DPOAE level at f2 frequencies 4kHz increased from birth to 6 months but then reduced slightly at 12 months. At lower frequencies the age-related DPOAE levels change was less significant. These results indicate that there are no major changes in DPOAE or cochlear function during the first year of life at most frequencies. However, the interpretation of DPOAE results in infants need to take into account the age-related difference in DPOAE pass rate at low-frequency and in DPOAE level at high-frequency. PMID- 17113743 TI - HPLC determination of four triterpenoids in rat urine after oral administration of total triterpenoids from Ganoderma lucidum. AB - A sensitive and simple high-performance liquid chromatography (HPLC) method was applied for the quantitative determination of four major triterpenoids (ganoderic acids C(2), B, K and H) in rat urine after oral administration of total triterpenoids from Ganoderma lucidum. The urine sample was extracted with dichloromethane-ethyl acetate (90:10) after acidification by hydrochloric acid (0.2 mol/ml). Chromatographic separation was achieved on a Zorbax SB-C(18) column (250 mm x 4.6 mm, 5 microm) at 35 degrees C, with a linear gradient of acetonitrile and 0.03% aqueous phosphoric acid (v/v), at a flow rate of 1.2 ml/min. The four triterpenoids and internal standard (hydrocortisone) were detected at a wavelength 252 nm. The within- and between-day assay coefficients of variation for the four triterpenoids in urine were less than 9% and the extraction recovery of this method was higher than 90%. Using this method, the excretion profile of the triterpenoids in rat urine after oral administration of total triterpenoids of G. lucidum was revealed for the first time. PMID- 17113744 TI - Effects of iodinated contrast media on endothelium: An in vitro study. AB - The aim of this study was to determine the influence of radiographic contrast media (CM) on endothelial cells in order to compare the effects of non-ionic (Iomeron and Visipaque) and ionic (Hexabrix and Uromiro) CM on the endothelial cells (EC). Human and murine cells were exposed for 2, 4 and 24h to increasing concentrations (12.5, 25, 50 and 100mg/mL) of test compounds. Controls were incubated with complete growth medium or mannitol solution (osmotic control). MTT assay was used to evaluate the cell viability, LDH assay was used to evaluate the membrane damage. The results demonstrate a difference between non-ionic and ionic compounds in the effect on endothelium. Ionic CM show to strongly affect endothelial cells viability under all tested conditions, while non-ionic CM show effects only after prolonged exposure at 50 and 100mg/mL, which represent instant concentrations lasting just minutes after intravascular injection of CM. Taken together, these results confirm that the currently employed non-ionic contrast media are well tolerated by the vascular endothelium and have wide margins of safety. PMID- 17113745 TI - Characterization and induction of human pre-adipocytes. AB - Human processed lipoaspirate is a source of multipotent adult stem cells that are able to differentiate between mesenchymal and neurogenic lineage. We characterized PLA cells by cytometry and then they were cultured to induce differentiation into myogenic and neurogenic lineage. Lipoaspirates were digested with collagenase to obtain the pellet, which was labelled with anti-CD44, anti CD45, and anti-CD90. We used BD FACS Calibur flow cytometer to acquire cellular events. Some cells were cultured at 37 degrees C and 5% CO(2) in neurogenic or myogenic medium and analysed by immunocytochemistry, using Neuron specific enolase, Vimentin, Glial fibrillary acidic protein, Tau, MAP2 to confirm neurogenic differentiation, MyoD1, Myosin heavy chain, Actin smooth muscle, vimentin to confirm myogenic differentiation. The cytometry results suggest that a part of the cells are of a mesenchymal origin, among which there are progenitor endothelial cells and leucocytes. Microscopy observation already reveals neuronal morphology and longitudinal multinucleated cells compared to control cells. Neurogenic cells only express NSE (early neuronal progenitor marker), but not GFAP, Tau, MAP2 (mature neuron and glial markers); myogenic cells are positive for MyoD1 and Myosin heavy chain. This demonstrates that lipoaspirate cells are capable of differentiating in vitro over a short period of time, and could be employed in biological and clinical research, like mesenchymal adult stem cells. PMID- 17113746 TI - Effect of mercury, cadmium, nickel, chromium and zinc on kinetic properties of NADPH-cytochrome P450 reductase purified from leaping mullet (Liza saliens). AB - Information on the mechanism of metal ion inhibition of NADPH-cytochrome P450 reductase is limited. The purpose of the present paper was to elucidate in vitro effect of Hg(+2), Cd(+2), Ni(+2), Cr(+3) and Zn(+2) ions on the purified mullet NADPH-cytochrome P450 reductase. NADPH-cytochrome P450 reductase was purified from detergent-solubilized liver microsomes from leaping mullet (Liza saliens). All of the metal ions caused inhibition of the enzyme activity except Zn(+2). At 50 microM metal concentration, Hg(+2) inhibited the cytochrome P450 reductase activity completely (100%), while, at the same concentrations, Cd(+2), Cr(+3) and Ni(+2) caused 66%, 65% and 37% inhibition, respectively. At 50 microM metal concentration, Zn(+2) had no apparent effect on cytochrome P450 reductase activity. The IC(50) values of HgCl(2), CrCl(3), CdCl(2) and NiCl(2) were estimated to be 0.07 microM, 24 microM, 33 microM and 143 microM, respectively. Of the metal ions tested, Hg(+2) exhibited much higher inhibitory effect at lower concentrations, so it was evidently a more potent inhibitor than the others. All four metal ions displayed noncompetitive type of inhibition mechanism for the purified reductase as analyzed by Dixon plot. K(i) values of Hg(+2), Cr(+3), Cd(+2), and Ni(+2) were calculated from Dixon plots as 0.048 microM, 18 microM, 73 microM and 329 microM, respectively. PMID- 17113748 TI - Possible therapeutic vaccines for canine myasthenia gravis: implications for the human disease and associated fatigue. AB - Myasthenia gravis (MG) is caused by T cell-dependent antibodies reactive with acetylcholine receptors. These autoreactive antibodies cause muscle weakness by interfering with neuromuscular transmission via removal of acetylcholine receptors from the neuromuscular junction as well as changing the architecture of the junction itself. Consequently, muscle fatigue is a debilitating aspect of MG often leading to more general feelings of tiredness not directly due to muscle weakness. We have previously described two peptides that are mimetics of antigen receptors on certain autoreactive T and B cells that are involved in MG. When used as vaccines in the rat model of MG, these peptides prevented and ameliorated disease and muscle fatigue by blunting acetylcholine receptor antibody responses. Such disease protection resulted from vaccine-induced anergizing antibodies against acetylcholine receptor-specific T and B cell antigen receptors. The present study prospectively evaluated the efficacy of these two vaccines in spontaneous acquired MG in pet dogs. When compared to historical controls that were prospectively studied, the vaccines increased the proportion of remitted dogs from 17 to 75%. In comparison to retrospectively studied historical controls that spontaneously remitted from MG, the vaccines accelerated the rate of decline in acetylcholine receptor antibody titers which resulted in a 3-fold decrease in the mean time to remission. These results are suggestive of a new type of targeted therapy that can drive autoimmune responses into long-term remission and possibly afford a means of determining whether correction of a physical cause of muscle weakness also corrects the perception of chronic, generalized fatigue. PMID- 17113749 TI - Host susceptibility to tuberculosis: CARD15 polymorphisms in a South African population. AB - Tuberculosis (TB) is one of the leading causes of death worldwide. The nucleotide binding oligomerisation domain 2 protein (NOD2) has recently been recognised as a non-redundant recognition mechanism of Mycobacterium tuberculosis. The caspase recruitment domain-containing protein 15 gene (CARD15), which encodes the NOD2 protein, is a susceptibility gene for Crohn's disease (CD), a granulomatous, chronic inflammatory disorder. CARD15 was therefore investigated as a candidate gene in TB. We genotyped the R702W, G908R and 1007fs variants, previously associated with CD, in TB cases and controls from the admixed South African Coloured population. No statistically significant differences between cases and controls were observed for these variants. We determined that the CD-associated mutations occur at very low frequencies in this population. Our results indicate that CARD15 is not a major susceptibility gene for TB in the South African Coloureds. PMID- 17113750 TI - Quantitative analysis of template-based attenuation compensation in 3D brain PET. AB - An atlas-guided attenuation correction method was recently proposed for 3D brain positron emission tomography (PET) imaging eliminating the need for acquisition of a patient-specific measured transmission scan. The algorithm was validated through comparison to transmission-based attenuation correction (gold standard) using voxelwise statistical parametric mapping (SPM) analysis of clinical data. In contrast to brain 'activation' studies for which SPM is primarily developed, brain PET research studies often involve absolute quantification. In the preliminary validation study published earlier, there is no validation as to how such quantification can be affected by the two methods as the assessment was carried out by an SPM group analysis alone. It is quite important to demonstrate how the proposed method performs individually, particularly for diagnostic applications or individual quantification. In this study, we assess the quantitative accuracy of this method in clinical setting using automated volume of interest (VOI)-based analysis by means of the commercially available BRASS software. There is a very good correlation (R(2)=0.91) between the atlas-guided and measured transmission-guided attenuation correction techniques and the regression line agreed well with the line of identity (slope=0.96) for the grouped analysis of patient data. The mean relative difference between the two methods for all VOIs across the whole population is 2.3% whereas the maximum difference is less than 7%. No proof of statistically significant differences could be verified for all regions. These encouraging results provide further confidence in the adequacy of the proposed approach demonstrating its performance particularly for research studies or diagnostic applications involving quantification. PMID- 17113751 TI - Mitogen-activated protein kinases, Erk and p38, phosphorylate and regulate Foxo1. AB - The members of the transcription factor Foxo family regulate the expression of genes concerned with the stress response, cell cycle and gluconeogenesis. Foxo1 (FKHR) contains 15 consensus phosphorylation sites for the mitogen-activated protein kinase (MAPK) family. Therefore, we hypothesized that MAPKs could directly regulate the transcriptional activity of Foxo1 via phosphorylation. In vitro kinase assay showed that Foxo1 was phosphorylated by extracellular signal regulated kinase (Erk) and p38 MAPK (p38) but not by c-jun NH2-terminal kinase (JNK). In NIH3T3 cells, epidermal growth factor or anisomycin increased phosphorylation of exogenous Foxo1, which was significantly inhibited by pretreatment with an MEK 1 inhibitor, PD98059, or a p38 inhibitor, SB203580. Two dimensional phosphopeptide mapping using mutation of phosphorylation sites for MAPK revealed that the nine serine residues in Foxo1 are specifically phosphorylated by Erk and that five of the nine residues are phosphorylated by p38 in vivo. Moreover, we also found that Foxo1 interacts with Ets-1 and functions as a coactivator for Ets-1 on the fetal liver kinase (Flk)-1 promoter in bovine carotid artery endothelial cells. Mutation of the nine phosphorylation sites for Erk in Foxo1 was shown to lead to less binding and synergistic activity for Ets-1 on the Flk-1 promoter when compared with wild-type Foxo1. These results suggest that Foxo1 is specifically phosphorylated by Erk and p38, and that this phosphorylation regulates the function of Foxo1 as a coactivator for Ets-1. PMID- 17113752 TI - Biological significance of dietary polyamines. AB - Polyamines are classically known by their names of putrescine, spermine, and spermidine. They are synthesized endogenously from ornithine and are interconvertible. In addition, an exogenous supply of polyamines is provided by dietary intake and by intestinal absorption from the products of bacterial metabolism. Polyamine uptake occurs almost entirely in the gut, and afterward the various forms are metabolized in different tissues under the strict regulation of ornithine decarboxylase, which is the first enzyme involved in their synthesis. Polyamines are eliminated from the organism by means of oxidation reactions, appearing in urine in all their metabolic forms. Polyamines play an important role in regulating cell growth and proliferation, the stabilization of negative charges of DNA, RNA transcription, protein synthesis, apoptosis, and the regulation of the immune response. They are components of breast milk and might be important in neonatal gut maturation, for which reason the possible supplementation of infant formulas with these compounds is under study. PMID- 17113753 TI - Serum levels of insulin-like growth factors (IGFs) and their binding proteins (IGFBPs), -1, -2, -3, in oral cancer. AB - INTRODUCTION: Insulin-like growth factor (IGF) possesses mitogenic properties promoting cellular proliferation and inhibiting cellular apoptosis. Reported evidence suggests that cancer (non-oral) is associated with high circulating levels of IGF-1 and low levels of IGFBP-3. AIMS: This study measured circulating levels of IFG-1 and IGFBPs -1, -2 and -3 in oral cancer patients. METHODOLOGY: Blood was collected from surgical patients and controls. Samples were assayed for IGF-1 and IGFBPs -1, -2, -3, using commercially available enzyme-linked immunosorbant assays (ELISA; R&D Systems Europe, Oxon, UK). RESULTS: Twenty-seven oral cancer and 31 age- and sex-matched patients were recruited. Mean IGF-1 and IGFBP-3 levels were significantly lower in cancer patients than controls (85.3 ng/ml and 2008 ng/ml versus 191 ng/ml and 2935 ng/ml, P<0.001). In contrast, levels of IGFBPs 1 and 2 were significantly higher in cancer patients than in controls. No significant association was demonstrated with tumour size or nodal metastases. DISCUSSION: This study has shown that in contrast to other cancers, circulating levels of IGF-1 and IGFBP-3 may both be lowered in patients with head and neck cancer. PMID- 17113754 TI - Comparison of three methods of facial measurement. AB - The aims of this study were to assess the accuracy of measurements recorded by 3D stereophotogrammetry and to compare three methods of facial measurement: manual anthropometry, 3D stereophotogrammetry and 2D photography. Measurements were taken from 14 landmarks on each of six volunteers and compared. In addition, the variability of the methods was assessed. Three-dimensional measurements were shown to compare well with manual measurements on volunteers as well as test objects for which the mean difference was 0.23 mm (shortest distance) and 0.13 mm (surface). All the three methods of measurement were found to have good levels of repeatability. Two-dimensional measurements were more variable than manual measurements (P=0.021). Three-dimensional stereophotogrammetric measurements were shown to compare well with manual measurements although the values obtained were mostly slightly larger. Stereophotogrammetry allows images to be taken in a Medical Photography Department, facilitating the accurate measurement of facial morphology from digitized data, including changes associated with treatment or growth. There are clear potential benefits of using 3D measurements in the assessment of facial deformity. PMID- 17113755 TI - Comparison of human alveolar osteoblasts cultured on polymer-ceramic composite scaffolds and tissue culture plates. AB - The effects of medical grade polycaprolactone-tricalcium phosphate (mPCL-TCP) (80:20) scaffolds on primary human alveolar osteoblasts (AOs) were compared with standard tissue-culture plates. Of the seeded AOs, 70% adhered to and proliferated on the scaffold surface and within open and interconnected pores; they formed multi-layered sheets and collagen fibers with uniform distribution within 28 days. Elevation of alkaline phosphatase activity occurred in scaffold cell constructs independent of osteogenic induction. AO proliferation rate increased and significant decrease in calcium concentration of the medium for both scaffolds and plates under induction conditions were seen. mPCL-TCP scaffolds significantly influenced the AO expression pattern of osterix and osteocalcin (OCN). Osteogenic induction down-regulated OCN at both RNA and protein level on scaffolds (3D) by day 7, and up-regulated OCN in cell-culture plates (2D) by day 14, but OCN levels on scaffolds were higher than on cell culture plates. Immunocytochemical signals for type I collagen, osteopontin and osteocalcin were detected at the outer parts of scaffold-cell constructs. More mineral nodules were found in induced than in non-induced constructs. Only induced 2D cultures showed nodule formation. mPCL-TCP scaffolds appear to stimulate osteogenesis in vitro by activating a cellular response in AO's to form mineralized tissue. There is a fundamental difference between culturing AOs on 2D and 3D environments that should be considered when studying osteogenesis in vitro. PMID- 17113756 TI - Deletion of distal promoter of VCXA in a patient with X-linked ichthyosis associated with borderline mental retardation. AB - BACKGROUND: X-linked ichthyosis (XLI) is caused by deficiency of steroid sulfatase (STS) activity. About 90% XLI patients have large deletions involving the entire STS gene and flanking regions. Recently, VCXA, which is located approximately 0.7Mb telomeric to the STS gene, was reported as a candidate gene for mental retardation (MR) in patients with XLI. OBJECTIVE: To delineate the X chromosomal deletion of a XLI patient with borderline mental retardation. METHODS: We carried out FISH analysis to show that the whole STS gene is deleted, and PCR analysis for fine-scale deletion mapping. RESULTS: The deleted segment is approximately 1.6Mb in size, and includes the entire STS and VCXB1 genes. VCXA itself is intact, but its promoter is deleted. CONCLUSION: A deletion that includes the VCXA promoter is associated with borderline mental retardation in a patient with XLI. PMID- 17113757 TI - Phenotypic and molecular characterisation of fish-borne Flavobacterium johnsoniae like isolates from aquaculture systems in South Africa. AB - Fish infections caused by pathogenic Flavobacterium species are a major problem in the aquaculture industry worldwide, often leading to large economic losses. Thirty-two Flavobacterium spp. isolates, obtained from various diseased fish species and biofilm growth, were characterised genetically using 16S rDNA PCR restriction fragment length polymorphism (RFLP), randomly amplified polymorphic DNA (RAPD) PCR, repetitive extragenic palindromic (REP) element PCR, plasmid profiling, whole cell protein (WCP) and outer membrane protein (OMP) analyses. Although the Flavobacterium spp. isolates displayed a high degree of genetic heterogeneity when differentiated by RAPD-PCR, REP-PCR and OMP fingerprinting techniques, isolates appeared very homogeneous by plasmid profiling and WCP analysis. No specific correlation was observed between the RAPD, REP and/or OMP profiles and fish host, site of isolation, geographic location or date of isolation of the Flavobacterium spp. isolates. Experimental infection of tilapia fish revealed variable levels of virulence and pathogenicity by isolates following handling stress and could not be linked to specific molecular types. This is the first reported isolation and characterisation of Flavobacterium johnsoniae-like spp. isolated from diseased fish in Southern Africa. PMID- 17113758 TI - Strain selection and improvement of gene transfer for genetic manipulation of Pseudomonas savastanoi isolated from olive knots. AB - Research on diseases of herbaceous plants caused by Pseudomonads has been rapidly progressing; however, for most pathovars which infect woody plants, strains accessible to genetic manipulation have not yet been reported. At present, few studies have reported the transfer of genes to Pseudomonas savastanoi pv. savastanoi, the causal agent of olive knot disease. A collection of P. savastanoi pv. savastanoi isolates was tested for its ability to receive, by conjugation, the broad-host range plasmid pBBR1MCS-2; four of them, showing conjugation frequencies higher than 10(-3) transconjugants/recipient, were selected. Differences in motility, colony size and morphology, and knot formation in olive explants were observed among the selected strains; nonetheless, amplification and sequencing of the 16S rRNA gene confirmed that they belonged to P. savastanoi species. Transformation frequency by electroporation of pBBR1MCS-2 into these strains was improved up to four orders of magnitude using plasmids isolated from a P. savastanoi strain and from an Escherichia coli modification/restriction deficient strain. Three of the selected strains maintained pBBR1MCS-2 stably and compatibly with their native plasmids during at least 90 generations, allowing the use of this vector for gene expression studies. Transposition via conjugation of different mini-Tn5, with or without the reporter genes gfp or luxAB, yielded frequencies varying from 1 x 10(-5) to 2.4 x 10(-9) transconjugants/recipient. Southern analysis of mutants obtained in strain NCPPB 3335 using a collection of DNA sequence tag transposons indicated that transposition occurs randomly, and in most cases at single sites in the genome of this strain, allowing the utilization of transposon tools for cell tagging and for the construction of insertional mutations. Knots developed on one-year-old plants inoculated with a Gfp-tagged strain clearly showed green fluorescence. PMID- 17113759 TI - A double-blind randomised comparative trial of amisulpride versus olanzapine for 2 months in the treatment of subjects with schizophrenia and comorbid depression. AB - PURPOSE: To compare the efficacy and safety of amisulpride and olanzapine in subjects with schizophrenia and comorbid depression in a randomised double-blind trial. PATIENTS: Eighty-five adult patients fulfilling DSM-IV criteria for schizophrenia and presenting a depressive episode were randomised to amisulpride (200-600 mg/day) or olanzapine (5-15 mg/day) for 8 weeks. Primary efficacy variables were change in Calgary Depression Scale (CDS) score and Clinical Global Impression (CGI) of Change. Safety was monitored by adverse event reporting and determination of extrapyramidal function and metabolic variables. RESULTS: The mean change from baseline of CDS score was -6.84 in the amisulpride group and 7.36 in the olanzapine group. 65.9% and 61.5% of subjects, respectively, were considered "much" or "very much" improved. No significant inter-group difference in effect size was observed. The frequency of adverse events was low and emergence of extrapyramidal symptoms was not seen. Four patients in the olanzapine group developed abnormal triglyceride levels. Mean weight gain was 1.45 and 0.5 kg, respectively, in the olanzapine and amisulpride groups. CONCLUSION: Amisulpride and olanzapine are effective in patients with schizophrenia and comorbid depression. Tolerance of both drugs was acceptable, although use of olanzapine was associated with a trend toward greater metabolic side-effects . PMID- 17113760 TI - Cdk5 is involved in NFT-like tauopathy induced by transient cerebral ischemia in female rats. AB - Although neurofibrillary tangle (NFT) formation is a central event in both familial and sporadic Alzheimer's disease (AD), neither cellular origin nor functional consequence of the NFTs are fully understood. This largely is due to the lack of available in vivo models for neurofibrillary degeneration (NFD). NFTs have only been identified in transgenic mice, bearing a transgene for a rare hereditary neurodegenerative disease, frontotemporal dementia with Parkinsonism linked to chromosome 17 (FTDP17). Epidemiological evidence suggests a much higher occurrence of dementia in stroke patients. This may represent the underlying cause of the pathogenesis of sporadic AD, which accounts for the majority of AD cases. We examined pathological markers of AD in a rodent stroke model. Here we show that after transient cerebral ischemia, hyperphosphorylated tau accumulates in neurons of the cerebral cortex in the ischemic area, forms filaments similar to those present in human neurodegenerative tauopathies and colocalizes with markers of apoptosis. As a potential underlying mechanism, we were able to determine that transient ischemia induced tau hyperphosphorylation and NFT-like conformations are associated with aberrant activation of cyclin dependent kinase 5 (Cdk5) and can be rescued by delivery of a potent, but non-specific cyclin dependent kinase inhibitor, roscovitine to the brain. Our study further indicates that accumulation of p35 and its calpain-mediated cleavage product, p25 may account for the deregulation of Cdk5 induced by transient ischemia. We conclude that Cdk5 may be the principal protein kinase responsible for tau hyperphosphorylation and may be a hallmark of the tauopathies in this stroke model. PMID- 17113761 TI - The ITPA c.94C>A and g.IVS2+21A>C sequence variants contribute to missplicing of the ITPA gene. AB - Inosine triphosphate pyrophosphatase (ITPase) catalyzes the conversion of inosine triphosphate (ITP) to the correspondent monophosphate. The ITPA c.94C>A and g.IVS2+21A>C allelic variants are associated with decreased red cell enzyme activity. The ITPA c.94C>A [P32T] sequence variant is associated with an increased risk of adverse drug reactions in patients treated with the thiopurine drug azathioprine. The aim of this study was to explore the molecular mechanisms of ITPase deficiency. ITPA mRNA was extracted from peripheral blood leukocytes (PBL), Epstein-Barr virus transformed lymphoblast cell cultures, reticulocytes, and cultured fibroblast from patients with known ITPA genotypes. ITPA mRNA was reversed transcribed, sequenced and the relative amounts of misspliced transcripts quantitated from three independent experiments. The ITPA g.IVS2+21A>C sequence variant resulted in missplicing of exon 3. The ITPA c.94C>A allelic variant resulted in missplicing of exons 2 and 3 representing, in PBL samples, 61% of the total mRNA expressed in ITPA c.94C>A homozygotes. We proposed that the ITPA c.94C>A allelic variant destroys an exonic splicing silencing (ESS) element in exon 2, resulting in the activation of two nearby upstream 5' splice sites and missplicing of the exons 2 and 3 cassette causing structural changes to the enzyme and contributing to ITPase deficiency. PMID- 17113762 TI - Functional nanofibrous scaffolds for bone reconstruction. AB - DNA compaction, encapsulation and stabilization strategies as well as a scheme for DNA chain stabilization by complex formation with modified fullerenes for gene delivery are discussed. DNA can be compacted in organic solvents and encapsulated with amphiphilic triblock copolymers. The rapid removal of the solvent mixtures by electrospinning together with a biodegradable polymer preserves the globular DNA conformation and can be used for bone reconstruction applications. Cationic fullerene surfactants can decorate and stabilize DNA coils in aqueous solution. The complex formation process is studied by static light scattering and analyzed in detail. PMID- 17113763 TI - Elevated neopterin levels in non-allergic asthma. AB - Neopterin is synthesized by human monocyte-derived macrophages upon stimulation with interferon-gamma (IFN-gamma). Measurement of neopterin concentration is useful to monitor cell-mediated (Th1-type) immune activation. In this study, we aimed to analyze the behaviour of neopterin in long lasting asthma considering its role as a marker of the Th1 environment and to establish the distinction between patients belonging either to the allergic or the non-allergic population, particularly in the elderly where asthma is often under diagnosed. Therefore we evaluated allergic parameters such as skin prick tests, IgE and hemogram (eosinophils count), and we compared our findings with neopterin values found in an age-matched control population. A group of individuals older than 65 was selected. It included 64 asthmatic patients (mean age 72+/-5 years) and 41 healthy individuals (mean age 79+/-7 years). In our study population, 42 patients presented positive skin tests, mainly to house dust mites. All patients were clinically stable and presented an average percentage of predicted forced expiratory volume in the first second (FEV1) of 73.6+/-25.3 and predicted median expiratory flow percentage (MEF50) of 38.8+/-26.7. Blood cell counts showed statistically different mean values of eosinophils between allergic and non allergic controls (5.42+/-4.7% versus 2.8+/-2.8%; p<0.04). IgE values were increased in allergic asthmatic patients when compared with non-allergic asthmatic patients (493.2+/-549.8IU/ml versus 85.3+/-194.4IU/ml; p=0.000). Allergic asthmatic patients presented mean neopterin levels similar to those found in the control group (2.4+/-2.8ng/ml versus 2.1+/-1.9ng/ml). In contrast, in non-allergic asthmatic patients these values were higher when compared with the control group (4.0+/-4.7ng/ml versus 2.1+/-1.9ng/ml). Neopterin levels were lower in allergic asthmatic patients when compared with non-allergic asthmatic patients (2.4+/-2.8ng/ml versus 4.0+/-4.7ng/ml). Within asthmatic patients, those with higher neopterin values (>2.1ng/ml) presented lower mean IgE values (IgE0.05) in either the peak heights of the GH response or the area under the curve (AUC) of the GH response after bGRF challenge on any of the four occasions of intensive bleeding. There were overall increases in plasma GH concentrations (P<0.01), AGR (P<0.01) and FCE (P=0.05) in the treatment group compared with the control animals. The study showed that GH responsiveness to administration of bGRF at 15-day intervals over 9 months of treatment remained unchanged in buffalo heifers. Exogenous bGRF treatment for a long period can therefore enhance GH release leading to higher growth rates and better feed conversion efficiency in buffalo heifers. PMID- 17113798 TI - The potential of oral vaccines for disease control in wildlife species. AB - Numerous infectious diseases caused by bacteria or viruses persist in developed and developing countries due to ongoing transmission among wildlife reservoir species. Such diseases become the target of control and management programmes in cases where they represent a threat to public health (for example rabies, sylvatic plague, Lyme disease), or livestock production (for example bovine tuberculosis, brucellosis, pseudorabies), or where they threaten the survival of endangered animal populations. In the majority of cases, lethal control operations are neither economically feasible nor publicly supported as a practical means for disease management. Prophylactic vaccination has emerged over the last 15 years as an alternative control strategy for wildlife diseases, mainly driven by the success of widescale oral rabies vaccination programmes for meso-carnivores in North America and Northern Europe. Different methods have been trialled for the effective delivery of wildlife vaccines in the field, however oral vaccination remains the most widely used approach. Successful implementation of an oral wildlife vaccine is dependent on a combination of three components: an efficacious immunogen, a suitable delivery vehicle, and a species-specific bait. This review outlines the major wildlife disease problems for which oral vaccination is currently under consideration as a disease management tool, and also focuses on the technological challenges that face wildlife vaccine development. The major conclusion is that attenuated or recombinant live microbes represent the most widely-used vaccines that can be delivered by the oral route; this in turn places major emphasis on effective delivery systems (to maintain vaccine viability), and on selective baiting systems, as the keys to wildlife vaccine success. Oral vaccination is a valuable adjunct or alternative strategy to culling for the control of diseases which persist in wildlife reservoirs. PMID- 17113799 TI - Blastic natural killer cell leukaemia in a dog--a case report. AB - A case of canine non-T, non-B lymphoid leukaemia was determined to be of natural killer (NK) cell lineage by detecting specific expression of canine CD56 mRNA by reverse transcriptase polymerase chain reaction analysis. Although NK cells are usually considered to be morphologically large granular lymphocytes, the malignant NK cells in this case were agranular and blast-like, resembling human blastic NK cell leukaemia. The prognosis of human NK cell leukaemia is usually poor. In this case, the dog died 10 days after initial presentation, despite chemotherapy. PMID- 17113800 TI - Enhanced resolution for EPR imaging by two-step deblurring. AB - The broad spectrum of spin probes used for electron paramagnetic resonance imaging (EPRI) result in poor spatial resolution of the reconstructed images. Conventional deconvolution procedures can enhance the resolution to some extent but obtaining high resolution EPR images is still a challenge. In this work, we have implemented and analyzed the performance of a postacquisition deblurring technique to enhance the spatial resolution of the EPR images. The technique consists of two steps; noniterative deconvolution followed by iterative deconvolution of the acquired projections which are then projected back using filtered backprojection (FBP) to reconstruct a high resolution image. Further, we have proposed an analogous technique for iterative reconstruction algorithms such as multiplicative simultaneous iterative reconstruction technique (MSIRT) which can be a method of choice for many applications. The performance of the suggested deblurring approach is evaluated using computer simulations and EPRI experiments. Results suggest that the proposed procedure is superior to the standard FBP and standard iterative reconstruction algorithms in terms of mean-square-error (MSE), spatial resolution, and visual judgment. Although the procedure is described for 2D imaging, it can be readily extended to 3D imaging. PMID- 17113801 TI - Experimentally induced cough. AB - The experimental induction of cough has become an important component of clinical cough research. Measurement of cough reflex sensitivity allows the evaluation of the effect of pharmacological and other interventions on the cough reflex, as well as the performance of epidemiological studies relevant to cough. The most commonly used tussive agents include capsaicin, citric acid and ultrasonically nebulized distilled water (fog). Methodological considerations are vital to the performance of high quality, reproducible cough challenge, which is especially important when measuring the effect of an experimental intervention on cough reflex sensitivity. For the optimal execution of clinical trials employing inhalation cough challenge, the subject population must be carefully selected, and the usefulness and potential limitations of data obtained from cough challenge studies need to be appreciated. PMID- 17113802 TI - Some vaguely explored (but not trivial) costs of tail autotomy in lizards. AB - Lizard tail autotomy is considered an efficient anti-predator strategy that allows animals to escape from a predator attack. However, since the tail also is involved in many alternative functions, tailless animals must cope with several costs following autotomy. Here we explicitly evaluate the consequences of tail autotomy for two costs that have been virtually unexplored: 1. we test whether the anatomical change that occurs after tail loss causes a reduction in the role of the tail as a distraction mechanism to predators; 2. we analyzed whether tail synthesis comprises an energetically costly process in itself, by directly comparing the cost of maintenance before and after autotomy. We found that original tails displace further and at greater velocity than regenerated tails, indicating that the anti-predation responses of a lizard probably changes according to whether its tail is original or regenerated. With regard to the energetic cost of tail synthesis, we observed a significant increase in the standard metabolic rate, which rose 36% in relation to the value recorded prior to tail loss. This result suggests that the energetic cost of tail synthesis itself could be enough to affect lizard fitness. PMID- 17113803 TI - Chemical characterization of the oligosaccharides in Bryde's whale (Balaenoptera edeni) and Sei whale (Balaenoptera borealis lesson) milk. AB - Samples of milk from a Bryde's whale and a Sei whale contained 2.7 g/100 mL and 1.7 g/100 mL of hexose, respectively. Both contained lactose as the dominant saccharide along with small amounts of Neu5Ac(alpha2-3)Gal(beta1-4)Glc (3'-N acetylneuraminyllactose), Neu5Ac(alpha2-6)Gal(beta1-4)Glc (6'-N acetylneuraminyllactose) and Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1 4)Glc (LST c). The dominance of lactose in the carbohydrate of these milks is similar to that of Minke whale milk and bottlenose dolphin colostrum, but the oligosaccharide patterns are different from those of these two species, illustrating the heterogeneity of milk oligosaccharides among the Cetacea. PMID- 17113804 TI - Serum insulin-like growth factor-I (IGF-I) IGF binding protein-3 (IGFBP-3) and leptin levels are related to abdominal aortic intima-media thickness in macrosomic newborns. AB - OBJECTIVE: Exposure to diabetes in utero has been established as a significant risk factor for some of the components of metabolic syndrome, and was associated with increased levels of maternal, placental, and fetal insulin-like growth factors and leptin. The atherogenic effects of leptin and insulin-like growth factor-I (IGF-I) have been extensively described. The present study was therefore designed to investigate relationships between abdominal aortic intima-media thickness (aIMT), serum IGF-I, IGF binding protein-3 (IGFBP-3) and leptin levels in macrosomic newborns. DESIGN: Neonates whose birth weights exceed 90th percentile for gestational age and gender are termed macrosomic. Abdominal aortic intima-media thickness was measured in 30 macrosomic neonates of diabetic mothers (group A), 30 macrosomic neonates of healthy mothers (group B) and 30 healthy neonates (group C). Serum IGF-I, IGFBP-3 and leptin levels were determined in all infants and their mothers. Stepwise logistic regression analysis was used to determine independent risk factors for aortic intima-media thickness. RESULTS: Mean aortic intima-media thickness was significantly higher in groups A and B (0.489+/-0.015,0.466+/-0.019 mm, respectively) than in controls (0.375+/-0.024 mm, p<0.0001). Weight-adjusted aortic intima-media thickness was significantly higher in-group A than in groups B (p=0.004) and C (p=0.048). Serum leptin concentration in-group B (37.4+/-10.7 ng/ml) was significantly greater than in group C (23.5+/-7.1 ng/ml, p<0.0001), but significantly lower than in-group A (46.6+/-14.1 ng/ml, p<0.0001). Serum IGF-I levels of the infants were significantly lower in-group C (113.2+/-33.1 ng/ml) than in groups A and B (205.2+/-60.1 and 179.3+/-55.1 ng/ml respectively, p<0.0001). Serum IGF-I, IGFBP 3 and leptin levels of the infants were positively correlated with mean (p<0.0001) and weight-adjusted aortic intima-media thickness measurements (p=0.003, p=0.006 and p=0.001, respectively). CONCLUSIONS: Macrosomic neonates of diabetic mothers have significantly increased aortic intima-media thickness with higher serum IGF-I, IGFBP-3 and leptin concentrations than those of controls. It might be speculated that these changes may exaggerate the atherosclerotic process later in life. PMID- 17113805 TI - Evolution of a Saccharomyces cerevisiae metabolic pathway in Escherichia coli. AB - The Saccharomyces cerevisiae glycerol pathway (GPD1 and GPP2) was evolved in vivo in Escherichia coli. The central metabolism of E. coli was engineered to link glucose consumption and glycerol production. The engineered strain was evolved in a chemostat culture and a high glycerol producer was rapidly obtained. The evolution of the strain was associated to a deletion between GPD1 and GPP2, resulting in the production of a fusion protein with both glycerol-3-P dehydrogenase and glycerol-3-P phosphatase activities. The higher efficiency of the fusion protein was due to partial glycerol-3-P channeling between the two active sites. The evolved strain produces glycerol from glucose at high yield, concentration and productivity. PMID- 17113806 TI - Mutation and biochemical analysis of 19 probands with mut0 and 13 with mut- methylmalonic aciduria: identification of seven novel mutations. AB - Isolated methylmalonic acidurias (MMA-urias) comprise a group of rare autosomal recessively inherited disorders characterised by accumulation of MMA in urine and other body fluids, resulting from deficient activity of the mitochondrial enzyme methylmalonyl-CoA mutase (MCM). Isolated MMA-uria results from either MCM apoenzyme defects (mut(0) and mut(-)) or defects in synthesis of its cofactor 5 deoxyadenosylcobalamin, i.e. cblA, cblB and cblD-variant 2. To date various studies have identified 171 disease-causing mutations in the MCM gene (MUT). We report mutation analysis in 32 probands with mut MMA-uria including 13 probands with a mut(-) defect. Sixty two of 64 possible mutant alleles were identified, seven of which were novel missense alleles. We found three novel mutations (c.427C>T/p.H143Y; c.862T>C/p.S288P; c.1361G>A/p.G454E) among 19 probands with a mut(0) defect and four novel mutations (c.299A>G/p.Y100C; c.1031C>T/p.S344F; c.1097A>G/p.N366S; c.2081G>T/p.R694L) among 13 probands with a mut(-) defect. Our study provides evidence that the p.Y100C, p.R108H, p.N366S, p.V633G, p.R694W, p.R694L and p.M700K mutations are associated with a mut(-) phenotype. PMID- 17113807 TI - Evaluation of adverse treatment effects in controlled trials. PMID- 17113808 TI - Polymyalgia rheumatica: diagnosis and treatment. AB - Polymyalgia rheumatica (PMR) typically manifests as inflammatory pain in the shoulder and/or pelvic girdles in a patient over 50 years of age. This condition was long underrecognized and therefore underdiagnosed. Today, however, overdiagnosis may occur. Physicians must be aware that many conditions may simulate PMR, including diseases that carry a grim prognosis or require urgent treatment. PMR may be the first manifestation of giant cell arteritis, and a painstaking search for other signs is mandatory. PMR may inaugurate other rheumatologic diseases such as rheumatoid arthritis, RS3PE syndrome, spondyloarthropathy, systemic lupus erythematosus (SLE), myopathy, vasculitis, and chondrocalcinosis. Finally, PMR may be the first manifestation of an endocrine disorder, a malignancy, or an infection. Failure to respond to glucocorticoid therapy should suggest giant cell arteritis, malignant disease, or infection. Ultrasonography may assist in the diagnosis by showing bilateral subdeltoid bursitis. Glucocorticoids are the mainstay of the treatment of PMR. Although the optimal starting dosage and tapering schedule are not agreed on, a low starting dosage and slow tapering may decrease the relapse rate. Methotrexate is probably useful when glucocorticoid dependency develops. In contrast, TNF alpha antagonists are probably ineffective. PMID- 17113809 TI - [Diagnosis of endometrial pathologies in West Africa: contribution of saline infusion sonography. Experience of Yopougon's teaching hospital (Abidjan, Ivory Coast)]. AB - OBJECTIVE: The aim of this survey was to value the contribution of saline infusion sonography (SIS) to endometrial pathologies diagnosis in a context of work where hysterography is the reference exam rather than hysteroscopy. PATIENTS AND METHODS: A prospective comparative cohort survey led at Yopougon's teaching hospital, from July 1st to April 30th 2003. This study concerned 65 patients. The sensitivity, the specificity, and the Kappa test have been calculated from the morbid results. RESULTS: The SIS has been successfully carried out among 63 patients (96,8%). The polyps (23,8%), and the mucous myomas (14%) dominate the endometrial pathologies. The analysis of the results indicated sensitivity and a global specificity respectively of 100% and 76,5%. The dissonant diagnoses have been decided in 82% of the cases in favour of SIS. Besides, SIS permits to save 31.67 Euros/patient. DISCUSSION AND CONCLUSION: SIS could therefore constitute a tool of choice for the exploration of the uterine cavity in the developing countries. PMID- 17113810 TI - [Are there any factors predicting failure or complications rates of trans obturator surgery for stress urinary incontinence?]. AB - OBJECTIVE: Since 2001 and the publication by Delorme of the trans-obturator route in the stress urinary incontinence (SUI), this technique has known an increasing development in France. The aim of this study is to evaluate the impact of different predicting factors on results and complications of trans-obturator surgery. PATIENTS AND METHODS: It is a retrospective, multicentric study, including 4 centers, 14 surgeons and 196 patients operated between February 2003 and August 2005. We have realized a univariate (Chi2 test) and multivariate (logistic regression test) statistic analysis concerning 7 sub-groups defined according to the literature on the TVT. RESULTS: Age>55 years (P=0,044) and SUI grade>2 (P=0,028) are statistically associated with a decrease of surgical success, age>55 years is also associated with an increase of complications rate in univariate (P=0,033) and multivariate (P=0,048) analysis. DISCUSSION AND CONCLUSION: Age>55 years should be considered, according to us, as a risk factor of surgical failure and complications in the trans-obturator surgery for SUI, none of the others risk factors found in the literature on the TVT seems to have an influence, in this study, on the results of trans-obturator surgery for SUI. PMID- 17113811 TI - [Prevalence of smoking cessation during pregnancy according to trimester]. AB - OBJECTIVE: To investigate the smoking cessation period during pregnancy. PATIENTS AND METHODS: Questionnaire-based, descriptive study of 979 pregnant women in four regions of France. The variables analysed included the characteristics of the mother and neonate at delivery, the smoking habits of the mother before and during pregnancy, the perception of risk linked to smoking, and the reasons for giving up smoking. RESULTS: Eighteen percent of women smoked until delivery. Forty-five percent of women gave up smoking during pregnancy, usually in the first trimester. More precisely, about one woman who smoked out of 50 gives up in order to prepare pregnancy. The proportion of women who stop smoking in each of the three trimesters of pregnancy is 84,1, 8,8 and 7,1% respectively. DISCUSSION AND CONCLUSION: Most women appear to stop smoking before any intervention therapy is possible. The first contact with a midwife or an obstetrician takes place whereas smoking cessation is already successful. PMID- 17113812 TI - FEM calculation of an acoustic field in a sonochemical reactor. AB - The spatial distribution of the acoustic amplitude in a sonochemical reactor has been numerically calculated using the finite element method (FEM). In the FEM program, the acoustic field in a sonochemical reactor is coupled with the vibration of the reactor's wall. The present calculations have revealed that the thin (thick) glass or stainless steel wall is nearly a free (rigid) boundary and that the glass wall is freer than the stainless steel wall. The influence of the attenuation coefficient of ultrasound on the acoustic field has also been studied in order to see the effect of bubbles on the acoustic field. As the attenuation coefficient increases, the vibration amplitude of the reactor's wall becomes smaller and the acoustic emission from the vibrating wall becomes weaker. The qualitative feature of the spatial pattern of sonochemiluminescence from an aqueous luminol solution has been reproduced by the calculation when the attenuation coefficient is in the range of 0.5-5m(-1). When the attenuation coefficient is less than about 0.05 m(-1), the standing wave pattern of an acoustic field in the liquid is very complex due to the acoustic emission from the vibrating wall. The present calculations have also revealed that some stripes of pressure antinodes have also been disconnected when the radius of the transducer is much smaller than the side length of the vibrating plate. The dependence of the acoustic field on the liquid height is also discussed. PMID- 17113813 TI - Immunohistochemical distribution of cyclic nucleotide phosphodiesterase (PDE) isoenzymes in the human vagina: a potential forensic value? AB - OBJECTIVES: Phosphodiesterase (PDE) isoenzymes are key proteins involved in the maintenance of the normal function of various tissues of the human body including those of the male and female urogenital tract. More recently, PDEs and their main substrates, cyclic GMP and cyclic AMP, have also been assumed to play a crucial role in the control of the human vagina. In order to elucidate the potential significance of phosphodiesterases as marker proteins in female genital organs, it was the aim of the present study to evaluate by means of immunohistochemistry the distribution of cGMP- and cAMP-PDE isoenzymes in specimens of the human vagina. METHODS: Conventional immunohistochemical techniques (double antibody technique, laser fluorescence microscopy) were applied to sections of the human vaginal wall in order to evaluate the presence of the PDE isoenzymes 1, 2, 3, 4, 5 and 10. RESULTS: Immunoreactivities (IR) specific for PDE1 (cAMP/cGMP-PDE, Ca(2+)/Calmodulin-dependent), PDE2 (cAMP-PDE, cGMP-dependent) and PDE5 (cGMP-PDE) were exclusively registered in the smooth musculature of vaginal arterial vessels, whereas no signals were detected in non-vascular tissue. IR indicating the expression of the cAMP-degrading PDE4 was mainly observed in the vaginal epithelium. Vaginal epithelial cells also presented immunosignals specific for PDE3 (cAMP-PDE, inhibited by cGMP) and PDE10 (dual substrate PDE), nevertheless, these stainings were less abundant than those related to the PDE4. IR for PDE10 was also registered in inflammatory cells located in the subepithelial region of the vaginal wall. CONCLUSION: Our study revealed the presence of IR specific for PDE1, PDE2, PDE4, PDE5 and PDE10 in sections of the human vagina and demonstrated that these enzymes are not evenly distributed in the tissue. Especially, the prominent expression of the cyclic AMP-PDE4A in the vaginal epithelium may give hint to a potential significance of this isoenzyme as a forensic marker protein. The findings give a rationale to investigate further as to whether the immunohistochemical detection of PDE4 may represent a new forensic tool in order to identify human vaginal epithelial cells. PMID- 17113814 TI - Phospholipids are synthesized in the G2/M phase of the cell cycle. AB - Very little is known about the metabolism of phospholipids in the G2 and M phases of the cell cycle, but limited studies have led to the postulation that phospholipid synthesis ceases during this period. To investigate whether phospholipids are synthesized in the G2/M phase of the cell cycle, protocols were developed to produce synchronized MCF-7 cell populations with greater than 80% of the cells in G1/S or G2/M phases that moved in synchrony following removal of the blocking agent. Analysis of the activities of key phosphatidylcholine and phosphatidylethanolamine biosynthetic enzymes in subcellular fractions obtained from MCF-7 cells at different cell cycle phases revealed that there was robust activity of key enzymes in the fractions prepared from MCF-7 cells in G2/M phase. Radiolabeled choline and ethanolamine were rapidly incorporated into cells maintained at G2/M phase with nocodazole, and the rates of incorporation were similar to those obtained in cells allowed to progress into the G1 phase. Furthermore, radiolabeled glycerol was incorporated into phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine and phosphatidic acid in MCF-7 cells maintained at G2/M phase with nocodazole. Similar results were obtained in CHO cells. These results demonstrate that glycerophospholipid synthesis is very active in the G2/M phase of these cells. Therefore, the postulated cessation of phospholipid synthesis in G2/M phases is not applicable to all cell types. PMID- 17113815 TI - The role of TNF-alpha in diabetic nephropathy: pathogenic and therapeutic implications. AB - Diabetes mellitus and its complications are a public health problem. Diabetic nephropathy has become the main cause of renal failure, and furthermore is associated with a dramatic increase in cardiovascular risk. Unfortunately, the mechanisms leading to the development and progression of renal injury in diabetes are not yet fully known. There is now evidence that activated innate immunity and inflammation are relevant factors in the pathogenesis of diabetes. Furthermore, different inflammatory molecules, including pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha), play a critical role in the development of microvascular diabetic complications, including nephropathy. This review discusses the role of TNF-alpha as a pathogenic factor in renal injury, focusing on diabetic nephropathy, and describes potential treatment strategies based on modulation of TNF-alpha activity. PMID- 17113816 TI - Sequencing single molecules of DNA. AB - In 2004, the NIH set a remarkable challenge: the 1000 dollars genome. Roughly speaking, success would provide, by 2015, the ability to sequence the complete genome of an individual human, quickly and at an accessible price. An intermediate goal of a 100,000 dollars genome was set for 2010. While the cost of Sanger sequencing has dropped dramatically over the past two decades, it is unlikely that the 100,000 dollars genome will be achieved by this means. New massively parallel technologies will push the cost of sequencing towards this mark, but it is doubtful whether these efforts will match the 1000 dollars goal. The best bets for ultrarapid, low-cost sequencing are single-molecule approaches. PMID- 17113817 TI - Report of the IWGT working group on strategy/interpretation for regulatory in vivo tests II. Identification of in vivo-only positive compounds in the bone marrow micronucleus test. AB - A survey conducted as part of an International Workshop on Genotoxicity Testing (IWGT) has identified a number of compounds that appear to be more readily detected in vivo than in vitro. The reasons for this property varies from compound to compound and includes metabolic differences; the influence of gut flora; higher exposures in vivo compared to in vitro; effects on pharmacology, in particular folate depletion or receptor kinase inhibition. It is possible that at least some of these compounds are detectable in vitro if a specific in vitro test is chosen as part of the test battery, but the 'correct' choice of test may not always be obvious when testing a compound of unknown genotoxicity. It is noted that many of the compounds identified in this study interfere with cell cycle kinetics and this can result in either aneugenicity or chromosome breakage. A decision tree is outlined as a guide for the evaluation of compounds that appear to be genotoxic agents in vivo but not in vitro. The regulatory implications of these findings are discussed. PMID- 17113818 TI - Effects of trematode double infection on the shell size and distribution of snail hosts. AB - Infection with larval trematodes sometimes alters the phenotypes of their snail hosts. While some trematode species have distinct effects on host phenotypes, it is still unclear how snail phenotypes are altered when they are parasitized with multiple trematode species. Here, we report that double infection with trematode species averages the effects of parasitic alteration on host phenotype. We found that snail hosts Batillaria attramentaria (Batillariidae) infected with Cercaria batillariae (Heterophyidae) have abnormally large shells and distribute in lower areas of the intertidal zone. Snails with another dominant trematode species, the renicolid cercaria I (Renicolidae), have slightly larger shells and distribute in upper areas of the intertidal zone. A number of double infections with both trematodes was observed in this study. Snails infected with both trematode species exhibited an intermediate size and inhabited a depth between those of snails solely infected with either trematode species, suggesting that the two trematodes simultaneously affected the snail phenotypes. Because altered host phenotypes are frequently beneficial to parasites, two trematode species may compete for successful transmission through alteration of host phenotypes. PMID- 17113819 TI - Spectrophotometric determination of gatifloxacin in pure form and in pharmaceutical formulation. AB - Simple, rapid, and extractive spectrophotometric methods were developed for the determination of gatifloxacin (GT) in bulk and pharmaceutical dosage form. These methods are based on the formation of yellow ion-pair complexes between the basic nitrogen of the drug and three sulphonphthalein acid dyes, namely; bromocresol green (BCG), bromocresol purple (BCP), bromophenol blue (BPB) and bromothymol blue (BTB) in phthalate buffer pH 3.0, 3.4 and 3.2, using BCG, BCP and (BPB or BTB), respectively. The formed complexes were extracted with chloroform and measured at 415, 417, 412 and 414 nm for BCG, BPB, BCP and BTB, respectively. The analytical parameters and their effects on the reported systems are investigated. The reactions were extremely rapid at room temperature and the absorbance values remains unchanged at 48 h for all reactions. Beer's law was obeyed in the ranges 2.0-20, 2.0-14 and 2.0-16 microg mL(-1) for BCG, BCP and (BPB or BTB), respectively. The composition of the ion pairs was found 1:1 by Job's method. Beer's law validation, accuracy, precision, limits of detection, limits of quantification. The proposed methods have been applied successfully for the analysis of the drug bulk form and its dosage form. The results were in good agreement with those obtained by the official and reported methods. PMID- 17113820 TI - Loss of characteristic absorption bands of C60 conjugation systems in the addition with aliphatic amines. AB - The characteristic absorption bands disappear and the shortest band at approximately 244 nm in cyclohexane or 282 nm in toluene remained only with long smoothing tail as C60 reacts adequately with aliphatic amines under sunlight radiation at approximately 40 degrees C. Simultaneously, fluorescence emission shifts from a weak band initially at longer wavelength to another strong one finally at shorter wavelength. The results might imply that the pi-conjugation system of C60 parent molecule is isolated into smaller separated parts. Therefore, some possible isolation models associated with observed experimental results are designed under some reasonable assumptive conditions. PMID- 17113821 TI - Multifrequency electron paramagnetic resonance study on deproteinized human bone. AB - Irradiated samples of deproteinized powdered human bone (femur) have been examined by electron paramagnetic resonance (EPR) spectroscopy in X, Q and W bands. In the bone powder sample only one type of CO2- radical ion is stabilized in the hydroxyapatite structure in contrast to powdered human tooth enamel, a material also containing hydroxyapatite, widely used for EPR dosimetry and in which a few radicals are stable at room temperature. It is suggested that the use of deproteinized bone for EPR dosimetry could improve the accuracy of dose determination. PMID- 17113822 TI - The molecular structure and vibrational spectra of 3-acetyl-4-[N-(2' aminopyridinyl)-3-amino]-3-buten-2-one by Hartree-Fock and density functional theory calculations. AB - The molecular structure and vibrational spectra of 3-acetyl-4-[N-(2' aminopyridinyl)-3-amino]-3-buten-2-one (C(11)H(13)N(3)O(2)) in the ground state have been investigated by Hartree-Fock and density functional method (B3LYP and BLYP) with 6-31G(d) basis set. The optimized geometric bond lengths and bond angles obtained by using HF and DFT show the best agreement with the experimental data. Comparison of the observed fundamental vibrational frequencies of title compound and calculated results by HF and DFT methods indicate that B3LYP is superior to the scaled HF approach for molecular problems. PMID- 17113823 TI - Infrared spectroscopy of racemic and enantiomeric forms of atenolol. AB - The molecular structure of conformational isomorphs given by X-ray diffraction for racemic and enantiomeric atenolol were optimized at the HF/6-31G* level of theory and the infrared spectra of the structure were calculated. These spectra are used to characterize the differences between the various atenolol conformers. The spectra of the (R,S)- and S-atenolol solid forms were recorded and the bands corresponding to the functional groups identified with the aid of the calculated spectra, fitting analysis, temperature effect and H/D isotopic exchange. Particular attention was paid to the stretch vibration modes of the functional groups present in the atenolol. PMID- 17113824 TI - Modeling in biomedical informatics: an exploratory analysis part 2. AB - OBJECTIVE: Modeling is a significant part of research, education and practice in biomedical and health informatics. Our objective was to explore which types of models of processes are used in current biomedical/health informatics research, as reflected in publications of scientific journals in this field. Also, the implications for medical informatics curricula were investigated. METHODS: Retrospective, prolective observational study on recent publications of the two official journals of the International Medical Informatics Association (IMIA), the International Journal of Medical Informatics (IJMI) and Methods of Information in Medicine (MIM). All publications of the years 2004 and 2005 from these journals were indexed according to a given list of model types. Random samples out of these publications were analysed in more depth. RESULTS: Three hundred and eighty-four publications have been analysed, 190 of IJMI and 194 of MIM. For publications in special issues (121 in IJMI) and special topics (132 in MIM) we found differences between theme-centered and conference-centered special issues/special topics (SIT) publications. In particular, we could observe a high variation between modeling in publications of theme-centered SITs. It became obvious that often sound formal knowledge as well as a strong engineering background is needed for carrying out this type of research. Usually, this knowledge and the related skills can be best provided in consecutive B.Sc. and M.Sc. programs in medical informatics (respectively, health informatics, biomedical informatics). If the focus should be primarily on health information systems and evaluation this can be offered in a M.Sc. program in medical informatics. CONCLUSIONS: In analysing the 384 publications it became obvious that modeling continues to be a major task in research, education and practice in biomedical and health informatics. Knowledge and skills on a broad range of model types are needed in biomedical/health informatics. PMID- 17113825 TI - We need more research into lung cancer. PMID- 17113826 TI - Role of phosphodiesterases in neurological and psychiatric disease. AB - Cyclic nucleotides such as cAMP and cGMP play a central role in neuronal cell function and are regulated by changes in synthesis and/or degradation. Degradation is regulated by phosphodiesterases (PDEs), a group of enzymes consisting of at least 11 families, several of which have multiple isoforms. As inhibition of PDEs can have profound effects on cell function, there is considerable interest in selective antagonists of these enzymes. Recent work has also revealed that PDEs are heterogeneously distributed, thus making them interesting targets for drug development. In particular, PDE10A may play a role in disorders that involve striatal neurons, such as Huntington's disease and psychosis. PMID- 17113827 TI - A niche for Wolbachia. AB - Wolbachia are endosymbionts of arthropods and filarial nematodes. Arthropods infected with these endobacteria display altered reproductive phenotypes, including cytoplasmic incompatibility and sex-ratio distortion. In nematodes, the endobacteria are essential for embryogenesis and worm survival. Wolbachia are transmitted vertically from mother to progeny, and Frydman et al. recently showed that, after transfer to uninfected Drosophila, Wolbachia rapidly accumulate in the somatic stem cell niche. From this location, the endobacteria might enter the developing oocytes and infect the progeny. PMID- 17113828 TI - Detection of antibodies reacting with the antithetical duffy blood group antigens Fy(a) and Fy(b) using recombinant fusion proteins containing the duffy extracellular domain. AB - Detecting blood group-specific antibodies in patient sera is essential to the management of blood transfusions or pregnancies. We produced the antithetical forms of the 65 amino acid extracellular domain (ECD) of the Duffy (Fy) blood group protein fused to glutathione sulfotransferase (GST): GST-Fy(a); and GST Fy(b), differing only in Gly or Asp at position 44, respectively. The purified recombinant proteins were recognized more effectively by reference polyclonal or monoclonal antibodies than the antithetical Fy specificity by either ELISA or immunoblotting. Combined immunoblot and ELISA tests performed at 1:200 dilutions of sera using the recombinant proteins gave results in agreement with undiluted sera and agglutination for 17/19 alloimmunized patients. At 1:200, agglutination detected anti-Fy(a) or anti-Fy(b) in only three of 12 samples that were positive by ELISA. Recombinant ECD-Fy proteins are suitable and sensitive reagents for the detection of anti-Fy that use technology amenable to automation and/or miniaturization and avoid the need for intact red cells. PMID- 17113829 TI - Detection of low avidity desmoglein 3-reactive T cells in pemphigus vulgaris using HLA-DR beta 1*0402 tetramers. AB - In the present study, we developed a HLA class II tetramer-based detection system utilizing DRB1*0402 tetramers loaded with recently identified immunodominant peptides of desmoglein 3 (Dsg3), the major autoantigen of pemphigus vulgaris (PV). Initial experiments demonstrated staining of a Dsg3-reactive T cell hybridoma which was derived from HLA-DR0402-transgenic mice with loaded PE labeled DRbeta1*0402 tetramers. However, staining of autoreactive T cell clones (TCC) derived from PV patients resulted only in positive staining by addition of exogenous peptides to the staining reactions. There was a dose-dependent specific binding of TCC to the tetramers with the agonistic Dsg3 peptide which was not altered by exogenous unrelated Dsg3 peptide. Noteworthy, the TCC did not stain with HLA-DR4 tetramers complexed with unrelated Dsg3 peptides. The findings of this study suggest that HLA class II tetramers may provide a highly specific approach to monitor ex vivo the T cellular autoimmune response against Dsg3 in patients with PV. PMID- 17113830 TI - Adrenoceptor and cholinoceptor modulation of rat pulmonary vein cardiac muscle contractility. AB - Cardiac muscle extends into mammalian pulmonary veins for variable distances according to species. This study has addressed the autonomic control of electrically paced cardiac muscle of the pulmonary vein of the rat. Contractile responses of Wistar rat pulmonary veins were investigated under isometric conditions in vitro. Vessels were electrically paced at 1 Hz (10 V, 1 ms pulse width). Acetylcholine (ACh, 1 nM-10 microM) attenuated the contractile response (maximum inhibition at 1 microM, 41+/-15%, mean+/-SD). The attenuation was inhibited by atropine (p<0.05) and partially inhibited (7+/-4%, mean+/-SD, p<0.01) by removal of the endothelium. Noradrenaline (NA, 1 nM-10 microM) augmented the cardiac muscle contractility in a fashion partially inhibited by atenolol; augmentation at 10 microM was reduced from 169+/-9% (n=6) to 135+/-9% (n=5), (p<0.05). The ability of ACh to attenuate the contractile responses was unaffected by the presence of NA. In conclusion, ACh has a muscarinic receptor mediated negative inotropic effect upon the cardiac muscle of the pulmonary vein of the rat mediated in part by the endothelium. The cardiac muscle expresses a positive inotropic response to NA partly mediated by beta1-adrenoceptors that can be antagonised by ACh. Therefore, pulmonary vein cardiac muscle function is modulated by competing autonomic influences which may be of significance to the generation of atrial fibrillation events. PMID- 17113832 TI - Infection, Genetics & Evolution: a journal with a high impact but no impact factor (as yet). PMID- 17113831 TI - The clinical trial of Women On the Move through Activity and Nutrition (WOMAN) study. AB - The Women On the Move through Activity and Nutrition (WOMAN) study is the first randomized clinical trial of nonpharmacological intervention designed to modify lipoproteins, weight loss and exercise among postmenopausal women using noninvasive measures of atherosclerosis as the primary endpoint. The trial was initially designed to test whether intervention as compared to health education would be more effective in slowing progression of subclinical atherosclerosis among women on hormone therapy (HT), estrogen or estrogen+progestin. It was designed and implemented prior to the results of the Women's Health Initiative (WHI). The trial was since modified to include women who had been on HT but went off after the results of the WHI were reported. Eligible women were between the ages of 52-62, had waist circumference>or=80 cm, low density lipoprotein cholesterol between 100-160 mg% and controlled blood pressure. The intervention is low in total and saturated fat, trans fats, higher in fiber and promotes loss of 7-10% of body weight and includes at least 150 min of physical activity per week. The study has recruited 508 women. The primary endpoints are change in extent of carotid intima-media wall thickness as measured by carotid ultrasound, pulse wave velocity as a measure of vascular stiffness and coronary artery calcium using electron beam computed tomography. Body composition is measured by dual-energy X-ray absorptiometry. PMID- 17113834 TI - Methods for samples preparation in proteomic research. AB - Sample preparation is one of the most crucial processes in proteomics research. The results of the experiment depend on the condition of the starting material. Therefore, the proper experimental model and careful sample preparation is vital to obtain significant and trustworthy results, particularly in comparative proteomics, where we are usually looking for minor differences between experimental-, and control samples. In this review we discuss problems associated with general strategies of samples preparation, and experimental demands for these processes. PMID- 17113833 TI - Hypersensitivity phenotypes associated with genetic and synthetic inhibitor induced base excision repair deficiency. AB - Single-base lesions in DNA are repaired predominantly by base excision repair (BER). DNA polymerase beta (pol beta) is the polymerase of choice in the preferred single-nucleotide BER pathway. The characteristic phenotype of mouse fibroblasts with a deletion of the pol beta gene is moderate hypersensitivity to monofunctional alkylating agents, e.g., methyl methanesulfonate (MMS). Increased sensitivity to MMS is also seen in the absence of pol beta partner proteins XRCC1 and PARP-1, and under conditions where BER efficiency is reduced by synthetic inhibitors. PARP activity plays a major role in protection against MMS-induced cytotoxicity, and cells treated with a combination of non-toxic concentrations of MMS and a PARP inhibitor undergo cell cycle arrest and die by a Chk1-dependent apoptotic pathway. Since BER-deficient cells and tumors are similarly hypersensitive to the clinically used chemotherapeutic methylating agent temozolomide, modulation of DNA damage-induced cell signaling pathways, as well as BER, are attractive targets for potentiating chemotherapy. PMID- 17113835 TI - Methods of analysis and separation of chiral flavonoids. AB - Although the analysis of the enantiomers and epimers of chiral flavanones has been carried out for over 20 years, there often remains a deficit within the pharmaceutical, agricultural, and medical sciences to address this issue. Hence, despite increased interest in the potential therapeutic uses, plant physiology roles, and health-benefits of chiral flavanones, the importance of stereoselectivity in agricultural, nutrition, pharmacokinetic, pharmacodynamic, pharmacological activity and disposition has often been ignored. This review presents both the general principles that allow separation of chiral flavanones, and discusses both the advantages and disadvantages of the available chromatographic assay methods and procedures used to separately quantify flavanone enantiomers and epimers in biological matrices. PMID- 17113836 TI - Preparation of membrane proteins for analysis by two-dimensional gel electrophoresis. AB - In order to separate hydrophobic membrane proteins, we have developed a novel two dimensional electrophoresis system. For the iso-electric focusing, agarose was used as a supporting matrix and n-dodecyl-beta-D-maltopyranoside was used as a surfactant. In combination with a previously developed Tris/MES electrophoresis system in the second dimension, distinct spots were reproducibly detected from hydrophobic membrane proteins whose grand average hydropathicity (GRAVY) exceed 0.3. In contrast to the immobilized pH gradient system, c-type heme was also visualized in this system. PMID- 17113837 TI - Purification and characterization of cassiicolin, the toxin produced by Corynespora cassiicola, causal agent of the leaf fall disease of rubber tree. AB - Cassiicolin, a phytotoxin produced by the necrotrophic fungus Corynespora cassiicola, was purified to homogeneity from a rubber tree isolate. The optimized protocol involves reverse phase chromatography followed by size exclusion chromatography, with monitoring of the toxicity on detached rubber tree leaves. Cassiicolin appeared to be a peptide composed of 27 amino acids, glycosylated on the second residue, with a N-terminal pyroglutamic acid and 6 cysteines involved in disulfide bonds. Its molecular mass was estimated to be 2885 Da. No significant sequence homology with other proteins could be found. The availability of pure toxin in sufficient amount is a prerequisite for its structure determination, which is a key step in the understanding of the aggression mechanism. PMID- 17113838 TI - HPLC columns partition by chemometric methods based on peptides retention. AB - In recent years, multivariate techniques have been utilized to evaluate reversed phase high-performance liquid chromatographic data. In the present study, 11 high performance liquid chromatography (HPLC) columns were divided into several groups according to the retention factors of 12 peptides. Principal component analysis (PCA) and cluster analysis (CA) were used in column and peptides' comparison and grouping. CA results indicated that all stationary phases may be generally grouped into several clusters, due to stationary phase structure and properties. On the other hand, interesting results were obtained with the use of PC. There is almost linear relationship between classified HPLC columns in the space of new PCs, which is connected with meaning of the PC's reflected in their loading values. The first component describes non-polar properties of peptides, whereas the second component is loaded with polar peptides having much lower logP values. PCA and CA were also used in peptides comparison however, complete explanation of peptides grouping still remains unclear. PMID- 17113839 TI - Simultaneous determination of ten antiarrhythic drugs and a metabolite in human plasma by liquid chromatography--tandem mass spectrometry. AB - A simple, accurate and selective LC-MS/MS method was developed and validated for simultaneous quantification of ten antiarrhythic drugs (diltiazem, amiodarone, mexiletine, propranolol, sotalol, verapamil, bisoprolol, metoprolol, atenolol, carvedilol) and a metabolite (norverapamil) in human plasma. Plasma samples were simply pretreated with acetonitrile for deproteinization. Chromatographic separation was performed on a Capcell C(18) column (50mmx2.0mm, 5microm) using a gradient mixture of acetonitrile and water (both containing 0.02% formic acid) as a mobile phase at flow rate of 0.3ml/min. The analytes were protonated in the positive electrospray ionization (ESI) interface and detected in multiple reaction monitoring (MRM) mode. Calibration curves were linear over wide ranges from sub- to over-therapeutic concentration in plasma for all analytes. Intra- and inter-batch precision of analysis was <12.0%, accuracy ranged from 90% to 110%, average recovery from 85.0% to 99.7%. The validated method was successfully applied to therapeutic drug monitoring (TDM) of antiarrhythic drugs in routine clinical practice. PMID- 17113840 TI - Simultaneous enantiomer determination of 20 (R)- and 20 (S)-ginsenoside-Rg2 in rat plasma after intravenous administration using HPLC method. AB - 20 (R,S)-Ginsenoside-Rg2, an anti-shock agent, is prescribed as a racemate. To analyze simultaneously the enantiomers of 20 (R)-ginsenoside-Rg2 and 20 (S) ginsenoside-Rg2 in plasma, a simple and reproducible high-performance liquid chromatographic (HPLC) method has been developed. The enantiomeric separation and determination were successfully achieved using a Diamonsil ODS C18 reversed-phase column (5 microm, 250 mm x 4.6 mm) with an RP18 (5 microm) guard column and a mobile phase of MeOH-aq. 4% H3PO4 (65:35, v/v, pH 5.1) with UV detection at 203 nm. Both enantiomers, 20 (R)-ginsenoside-Rg2 and 20 (S)-ginsenoside-Rg2, were well separated at 14.5 min and 13.6 min, respectively. The linear ranges of the standard curves were 2.0-250 microg/ml. The intra- and inter-day precision (R.S.D.) were 0.05). The relative mortality risk within 30 days of RP was approximately nine times the baseline risk (95% confidence interval 3 to 38) and was similar for men in all three age groups (P >0.05). CONCLUSIONS: The results of our study indicate that men aged 70 to 79 years do not have a greater absolute excess or relative risk of 30-day mortality after RP compared with men aged 60 to 69 years. PMID- 17113898 TI - Prospective health-related quality-of-life assessment in an initial cohort of patients undergoing robotic radical prostatectomy. AB - OBJECTIVES: To prospectively assess the health-related quality-of-life outcomes of patients undergoing robot-assisted laparoscopic prostatectomy using a validated patient self-assessment questionnaire. METHODS: Patients undergoing robot-assisted laparoscopic prostatectomy between September 2003 and May 2005 were given the Expanded Prostate Cancer Index Composite questionnaire preoperatively and 1, 3, 6, 9, 12, and 18 months postoperatively. Patients with a minimum follow-up of 3 months were included in the analysis. The mean domain specific health-related quality-of-life scores +/- SD and the proportion of patients achieving their baseline scores were calculated. Multivariate proportional hazards regression analysis was used to determine the potential prognostic factors for a return to baseline of the domain scores and continence. RESULTS: The median follow-up was 9.5 months. The median time to recovery of the baseline summary scores was 6.6 months (95% confidence interval [CI] 5.9 to 7.2) in the urinary domain, 2.8 months (95% CI 2.0 to 3.7) in the bowel domain, and 3.0 months (95% CI 2.2 to 3.9) in the hormonal domain. The baseline sexual summary score was recovered by 19.2% of patients at 12 months. The median time to return of continence (0 to 1 pads/day) was 4.0 months (95% CI 3.0 to 4.9). The median time to the return of erections firm enough for intercourse was 13.5 months (95% CI 9.9 to 17.1). On multivariate proportional hazards regression analysis, age, body mass index, prostate size, nerve-sparing technique, and number of comorbidities were not significantly associated with the time to recovery of the baseline domain scores or continence. CONCLUSIONS: Patients undergoing robot-assisted laparoscopic prostatectomy have a favorable health related quality-of-life recovery profile that appears comparable to those of established surgical approaches. PMID- 17113899 TI - Intravesical injection of botulinum toxin type A: management of neuropathic bladder and bowel dysfunction in children with myelomeningocele. AB - OBJECTIVES: To evaluate the efficacy of botulinum toxin A in the treatment of children with detrusor hyperreflexia caused by myelomeningocele and the effects of this treatment on neuropathic bladder and bowel dysfunction. METHODS: In a prospective study, 26 children with myelomeningocele (20 boys and 6 girls, mean age 6.9 years) were included. All patients had been nonresponders to medical treatment and required clean intermittent catheterization. Under cystoscopic guidance, 10 IU/kg of botulinum toxin A was injected into the detrusor muscle, sparing the trigone and ureteral orifices. In each patient, urinary incontinence grade and improvement in parameters of interest in the evaluation of bowel dysfunction were assessed before and 4 months after injection. Conventional urodynamic studies to determine maximal bladder capacity and maximal detrusor pressure and voiding cystoureterography were also performed. RESULTS: Four months after procedure, 19 patients (73%) had become completely dry between clean intermittent catheterizations, and the total improvement in urine incontinence was 88%. The mean maximal detrusor pressure was decreased to 83.2 +/- 4.6 cm H2O from the baseline of 139.3 +/- 11.2 (P <0.01). The average maximal bladder capacity increased from 102.8 +/- 6.3 mL to 270.2 +/- 9.5 mL (P <0.01). Of the 15 patients who had varying degrees of vesicoureteral reflux before the procedure, 11 (73%) had decrease in the vesicoureteral reflux grade. Also, bowel dysfunction improved in 10 (66%) of the 15 patients. CONCLUSIONS: Botulinum toxin A appears to be a safe, minimally invasive procedure for the management of neuropathic bladder and bowel dysfunction in children with myelomeningocele. PMID- 17113902 TI - Development of nocturnal urinary control in Chinese children younger than 8 years old. AB - OBJECTIVES: To estimate the development of nocturnal urinary control (NUC) with age in Chinese children younger than 8 years of age using cross-sectional and retrospective surveys. METHODS: We used a cross-sectional survey of 4754 children (1 to 8 years old), a retrospective investigation of 2745 children younger than 9 years old, and an anonymous questionnaire of 8222 children 9 to 18 years old . The children without NUC were subdivided into infant (1 to 3 years old), preschool age (4 to 6 years old), and primary school (7 to 8 years old) groups. RESULTS: The response rate to the cross-sectional and retrospective surveys was 90% and 89%, respectively. In the cross-sectional survey, the prevalence of children attaining NUC was 52% for those younger than 2 years of age, 76% for those aged 2 to 3 years, and 93% at age 8. Girls were more likely than boys to acquire NUC earlier. In the retrospective survey, the prevalence of children attaining NUC before age 2 was 17% and was 72% for those aged 2 to 3 years, and 98% by age 8. The proportion of nonmonosymptomatic bedwettings in children without NUC was 14%. Arousal difficulty and a positive family history were found in 67% and 11% of children with nocturnal wetting, respectively. The severity of bedwetting and arousal difficulty was significantly greater in infants than in preschool and school-age children. CONCLUSIONS: The results from the cross sectional and retrospective surveys showed that the most important period of attaining NUC is 2 to 3 years of age. Nearly 90% of children attained NUC by the age of 5. PMID- 17113905 TI - An efficient solution to the retained Foley catheter. AB - The most common cause of a retained Foley catheter is failure of the balloon to deflate. If noninvasive means are unsuccessful, balloon puncture is required. We present a safe and efficient technique using flexible video cystoscopy for balloon puncture and removal of any free fragments formed. PMID- 17113906 TI - Adrenocortical carcinoma with renal vein invasion in a woman with a horseshoe kidney. PMID- 17113907 TI - Unusual case of bladder duplication: complete duplication in coronal plane with single urethra and no associated anomalies. AB - Duplication of the bladder is an unusual congenital anomaly. Complete and incomplete forms are well defined in published reports. We present an unusual case of a 3-year-old boy with two separate-walled bladders, one lying in front of the other, communicating through a tiny septum, at the neck of the primary bladder that drained to a normal single urethra. This type of isolated bladder anomaly was not defined in the classification of the duplication of the bladder. In patients with a lower abdominal cystic mass, complete bladder duplication must be included in the differential diagnosis of pelvic cystic masses. PMID- 17113908 TI - Use of positron emission tomography in spindle cell carcinoma of the penis. AB - A 60-year-old man presented with spindle cell carcinoma of the penis. He underwent surgery and additional positron emission tomography and computed tomography scans to evaluate for possible metastases. Positron emission tomography showed a left inguinal and paravesical hot spot on the right. Only the left inguinal lesion could be confirmed on computed tomography. The patient underwent additional surgery with curative intent. Three months later, the patient underwent repeat computed tomography, which revealed an osteolytic process in the right acetabulum. This lesion corresponded with the right paravesical hot spot on the positron emission tomography scan 3 months earlier. PMID- 17113909 TI - Absence of Gerota's fascia in pelvic ectopic kidney: implications in laparoscopic radical nephrectomy. AB - We report the anatomic alterations in pelvic ectopic kidney and its bearing on the performance of laparoscopic radical nephrectomy (LRN) and its likely effect on the ultimate oncologic outcome. A patient with Stage T2N0M0 renal tumor in a pelvic kidney underwent transperitoneal LRN. LRN was performed by mobilizing the specimen with an adequate amount of perirenal fat, because no distinct Gerota's fascia can be found in the pelvic kidney. At 1 year of follow-up, the patient was free of disease. LRN is a good option for managing localized renal tumor even in the pelvic kidney. The impact of the absence of Gerota's fascia in the pelvic kidney needs further evaluation. PMID- 17113910 TI - Partial nephrectomy using a monopolar radiofrequency device: an animal model. AB - OBJECTIVES: To compare the use of radiofrequency (RF) and electrocautery in partial nephrectomy without renal artery clamping for bleeding and tissue destruction. METHODS: Sixteen adult rabbits were randomized into two groups. Partial nephrectomy using a monopolar RF device without renal artery clamping was performed in 8 rabbits (RF group) and partial nephrectomy using electrocautery was performed in the rest (cautery group). Four rabbits in each group (rabbits 1, 3, 5, and 7) were kept for follow-up, and the operated kidneys of the rest were removed for histopathologic evaluation. The tissue samples were placed in 10% formalin solution and sent to the pathology laboratory. The groups were compared in terms of bleeding and tissue destruction. RESULTS: The mean blood loss was 3.6 +/- 1.2 mL in the RF group and 8.3 +/- 2.7 mL in the control group (P = 0.003). A rabbit in the control group died on postoperative day 3 because of bleeding. Others were followed up for 3 months postoperatively. The amount of thermal destruction was comparable between the two groups. Varying degrees of thermal destruction were observed at the cutting margins in both groups. No difference was found between the two groups in terms of the deepness of thermal injury (1 to 2 mm). CONCLUSIONS: The findings of our experimental study showed that the use of the RF electrode in partial nephrectomy without renal artery clamping resulted in less intraoperative bleeding without differences in terms of tissue destruction. PMID- 17113911 TI - Participation of adrenomedullin and its relation with vascular endothelial growth factor in androgen regulation of prostatic blood flow in vivo. AB - OBJECTIVES: We had previously reported that androgen-regulated prostatic blood flow and vascular endothelial growth factor (VEGF) were involved in the signal transduction pathway. Adrenomedullin (ADM) is a multifunctional regulatory peptide with mitogenic and angiogenic capabilities that are regulated by androgen. ADM is abundantly expressed in the prostate. We focused on ADM and evaluated its participation and relation with VEGF in androgen prostatic blood flow regulation using a castrated rat model. METHODS: We examined the effect of locally injected dihydrotestosterone (DHT) and ADM, and the co-administration of DHT with an ADM receptor antagonist (ADM 22-52) on prostatic blood flow. Furthermore, prostatic blood flow was evaluated after ADM and VEGF administration with each other's antagonist, VEGF neutralizing antibody and ADM 22-52, respectively. Changes in the mRNA expression levels of ADM in the prostate after castration and successive androgen stimulation were also evaluated. RESULTS: The administration of ADM promptly increased prostatic blood flow in a dose-dependent manner within 30 minutes. The DHT-induced increase in prostatic blood flow was completely abolished by co-administration with anti-ADM. Anti-ADM inhibited the VEGF-induced prostatic blood flow elevation, but a VEGF neutralizing antibody did not affect the ADM-mediated blood flow elevation. Furthermore, upregulation of the ADM gene induced by DHT was inhibited by co-administration with a VEGF neutralizing antibody. CONCLUSIONS: These results have clearly demonstrated the direct regulation of prostatic blood flow by ADM and its involvement in androgenic prostatic blood flow regulation. Furthermore, ADM was estimated to be a downstream mediator of VEGF action in the signal transduction pathway. PMID- 17113912 TI - Alteration of contractile and regulatory proteins in estrogen-induced hypertrophy of female rabbit bladder. AB - OBJECTIVES: Estrogen is essential to mediate physiologic functions in female bladders. Deficiency of estrogen has been speculated to be an etiologic factor for bladder dysfunction in postmenopausal women. Our previous studies have demonstrated that estrogen supplementation in female rabbits induces a "functional hypertrophy" of the urinary bladder smooth muscle. The present study investigated the alterations in the contractile and regulatory proteins in this model. METHODS: Twenty New Zealand white female rabbits were separated into five groups of 4 rabbits each. Group 1 served as the control, groups 2 to 6 underwent ovariectomy (Ovx), and group 2 served as the Ovx without estradiol treatment group. Two weeks after Ovx, groups 3 to 5 were given 17-beta estradiol (1 mg/kg/day) by subcutaneous implant for 1, 3, and 7 days, respectively. The expression of the contractile and regulatory proteins, such as myosin light chain kinase, rho-kinase, and caldesmon, was analyzed by Western blotting. RESULTS: The expression of myosin light chain kinase was enhanced by estradiol supplementation. The expression of rho-kinase-alpha was increased significantly (20-fold) after Ovx, which was downregulated after estrogen supplementation. No significant change was seen in rho-kinase-beta after Ovx or estradiol supplementation. The expression of caldesmon isoforms was enhanced by 1-day estradiol supplementation but decreased to lower levels than those of the control group by 3 and 7 days of estrogen treatment. CONCLUSIONS: The results of the present study have provided more understanding about the role of the contractile and regulatory proteins in detrusor muscle, in both dysfunctional atrophy induced by Ovx and functional hypertrophy induced by estrogen supplementation. PMID- 17113913 TI - Mannose-binding lectin binds IgM to activate the lectin complement pathway in vitro and in vivo. AB - Recent evidence has implicated a role for the MBL-dependent lectin pathway in gastrointestinal and myocardial ischemia/reperfusion (I/R)-induced injury. However, previous studies have implicated IgM and the classical pathway as initiators of complement activation following I/R. Thus, we investigated the potential interaction between MBL and IgM leading to complement activation. Using surface plasmon resonance, we demonstrate that MBL does bind human IgM. Subsequently, functional complement activation was demonstrated in vitro following sensitization of human RBCs with mouse anti-human CD59 IgM and more lysis was observed with MBL sufficient sera compared to MBL deficient (KO) sera. Similarly, treatment of human endothelial cells with mouse anti-human CD59 IgM, MBL and MASP-2 activated and deposited C4. These data suggest that the presence of both IgM and MBL can activate the lectin pathway in vitro. Serum ALT levels increased significantly in sIgM/MBL-A/C KO mice reconstituted with WT plasma compared to sIgM/MBL-A/C KO mice reconstituted with MBL-A/C KO plasma following gastrointestinal (G) I/R. Similarly, intestinal C3 deposition was greater in sIgM/MBL-A/C KO mice reconstituted with WT plasma compared to sIgM/MBL-A/C KO mice treated with MBL-A/C KO plasma. These data indicate for the first time that both IgM and MBL-A/C are required for GI/R-induced complement activation and subsequent injury. PMID- 17113914 TI - Synthetic triacylated lipid A derivative activates antigen presenting cells via the TLR4 pathway and promotes antigen-specific responses in vivo. AB - Triggering the maturation of dendritic cells (DC) with toll-like receptor (TLR) agonists is a favored strategy for the development of vaccine adjuvants. The triacyl pseudo-dipeptidic agent OM-197-MP-AC mimicking the lipid A structure of endotoxin induces the maturation of human monocyte-derived DC. In this study we investigated the signaling pathway by which this molecule activates DC. The ability of OM-197-MP-AC to induce maturation of human and mouse DC and macrophages was dependent on TLR4, not TLR2. Ovalbumin-specific humoral and T helper cell responses were significantly augmented by OM-197-MP-AC treatment. Taken together these results indicate that OM-197-MP-AC is a TLR4 agonist inducing DC maturation and represents a novel class of vaccine adjuvants devoid of the known pyrogenic effects associated with classical LPS derivatives. PMID- 17113915 TI - Calcium-pterin suppresses mitogen-induced tryptophan degradation and neopterin production in peripheral blood mononuclear cells. AB - Antitumor activity of a calcium-pterin suspension has been described in vitro and in animal model systems. Recent studies provide some evidence that this effect involves immune-mediated mechanisms. We investigated the influence of calcium pterin on freshly isolated human peripheral blood mononuclear cells (PBMC) stimulated with the mitogens phytohaemagglutinin and concanavalin A in vitro. Influence of calcium-pterin on tryptophan-degrading enzyme indoleamine (2,3) dioxygenase (IDO) and on neopterin production was monitored in supernatants of cells. Increased neopterin concentrations as well as accelerated tryptophan degradation have been found to predict poor prognosis in patients with cancer, and both these immunobiochemical pathways are induced by the pro-inflammatory cytokine interferon-gamma. Compared to unstimulated cells, mitogens induced degradation of tryptophan and formation of neopterin in PBMC, and upon addition of calcium-pterin, both biochemical results were suppressed in a dose-dependent way. Thus, calcium-pterin suppresses immunological pathways in vitro that in patients with malignant diseases characterize an unfavorable prognosis. The effect of the compound to suppress IDO activity could be of considerable relevance for the antitumoral effect of the compound because activation of the enzyme is considered as an immune-escape mechanism of tumor cells. PMID- 17113916 TI - Low glucocorticoid receptor (GR), high Dig2 and low Bcl-2 expression in double positive thymocytes of BALB/c mice indicates their endogenous glucocorticoid hormone exposure. AB - Several studies have shown that of the four major thymocyte subsets, the CD4/CD8 double positive (DP) thymocytes are the most sensitive to in vivo glucocorticoid hormone (GC)-induced apoptosis. Our aim was to analyse fine molecular differences among thymocyte subgroups that could underlie this phenomenon. Therefore, we characterised the glucocorticoid hormone receptor (GR) expression of thymocyte subgroups both at the mRNA and protein levels by real-time PCR and flow cytometry, and correlated these features to their apoptotic sensitivity. We also investigated the time-dependent effects of the GC agonist dexamethasone (DX) with or without GC antagonist (RU486) treatments on GR mRNA/protein expression. We also analysed the expression of two apoptosis-related gene products: dexamethasone-induced gene 2 (Dig2) mRNA and Bcl-2 protein. We found that DN thymocytes had the highest GR expression, followed by CD8 single positive (SP), CD4 SP and DP thymocytes in 4-week-old BALB/c mice, both at the mRNA and protein levels, respectively. In DP cells, the Dig2 expression was significantly higher, while the Bcl-2 expression was significantly lower than in DN, CD4 SP and CD8 SP thymocytes. Single high dose DX treatment caused time-dependent depletion of DP thymocytes due to their higher apoptosis rate, which could not be abolished with RU486 pretreatment. After a single high dose DX treatment, there was a transient, significant increase of the GR mRNA and protein level of unsorted thymocytes after 8 and 16 h, followed by a significant decrease at 24 h, respectively. The time-dependent GR expression changes after DX administration could not be inhibited by the GC antagonist RU486. Twenty-four hours after exposure to high dose DX the DN, CD4 SP and CD8 SP cells showed a significant decrease of GR mRNA and protein expression, whereas the DP thymocytes, showed no significant alteration of GR mRNA or protein expression. The kinetical analysis of GR expression and apoptotic marker changes upon single high dose GC analogue administration revealed a two-phase process in thymocytes: early events, within 4 8 h, include GR upregulation and early apoptosis induction, while the late events appear most prominently at 16-20 h, when the GR is already downregulated and apoptotic cell ratio reaches its peak, with marked DP cell depletion. The low GR, high Dig2 and low Bcl-2 expression, coupled with the absence of homologous downregulation of GR after exogenous GC analogue treatment, could contribute to the high GC sensitivity of DP thymocytes. The downregulated GR and Bcl-2 together with the upregulated Dig2 level in DP cells indicates the significance of intrathymic GC effects at this differentiation stage. Since GR expression changes and apoptotic events could not be completely inhibited by GC antagonist, we propose the involvement of non-genomic GR mechanisms in these processes. PMID- 17113917 TI - Development of diagnostic reagents: raising antibodies against synthetic peptides of PfHRP-2 and LDH using microsphere delivery. AB - Malaria causes significant morbidity and mortality worldwide, including countries with mainly imported malaria. In developing nations, scarce resources lead to inadequate diagnostic procedures. Microscopy of Giemsa-stained thick and thin films remains the current gold standard for diagnosis. Although it has good sensitivity and allows species identification, it is time consuming, requires microscopical expertise and maintenance of equipment. Antigen tests are promising tools for the diagnosis of malaria. Two such antigens are Plasmodium falciparum histidine rich protein (pfHRP-2) and lactate dehydrogenase (LDH). The present study was aimed to develop indigenous, rapid and sensitive immunodiagnostic method based on detection of PfHRP-2 and LDH antigen in the blood. Unique peptide sequences of PfHRP-2 (two regions) and LDH (three regions) antigen were synthesized by solid phase technique and purified to homogeneity. The antibodies raised against these sequences were raised in mice as well as rabbit using microspheres (PLGA) to generate high titre and affinity antibodies. The peptide specific peak titres varied from 25,000 to 50,000 and affinity of the antibodies produced was found to be in order of 0.73-5.3 nM. The antibodies generated using microspheres were able to detect the PfHRP-2 and LDH antigen in the culture supernatant and parasitized RBC lysate of P. falciparum respectively by sandwich ELISA up to 0.002% parasitaemia level. The assay allowed the detection of parasite infections of 0.08-2.68% parasitaemia with a sensitivity of 100% in the whole blood of P. falciparum positive patients. No cross-reactivity was observed with P. vivax infected patients. PMID- 17113918 TI - The superantigen staphylococcal enterotoxin A (SEA) and monoclonal antibody L243 share a common epitope but differ in their ability to induce apoptosis via MHC II. AB - Crosslinking of MHC class II (MHC-II) molecules by antibodies or by superantigens (SAg) induces a variety of functional responses in the antigen presenting cell. We were able to allocate K39 as the residue that is essential for binding of antibody L243 to the alpha chain of HLA-DR. K39 is also essential for binding of staphylococcal enterotoxin A (SEA). However, the functional responses of the two ligands differ considerably exemplified by the ability of L243 to induce apoptosis in monocytic cells and in B cells, whereas SEA is unable to activate the apoptosis pathway. Despite the differences in functional responses, both ligands induce cell aggregation in MonoMac-1 cells. The SEA molecule with its two different binding sites associates one MHC alpha chain with one beta chain as opposed to two alpha chains that are brought into close proximity by the two identical antigen binding sites of L243. We therefore conclude that the spatial orientation of dimerized MHC-II and their associated proteins is an important factor for the nature of the transduced signal and consequently the outcome of functional responses. PMID- 17113919 TI - Inflammatory signal transduction from the Fc epsilon RI to NF-kappa B. AB - Mast cells are essential effector cells in IgE-associated immune responses. The major receptor for mast cell activation is the high affinity IgE receptor Fc epsilon RI. Fc epsilon RI crosslinking induces mast cell degranulation and de novo synthesis of potent proinflammatory mediators. Recent work identified Bcl10 and Malt1 as central regulators of a specific signaling pathway that controls NF kappaB activation and proinflammatory cytokine production upon Fc epsilon RI ligation on mast cells. Bcl10 and Malt1 cooperate for the activation of this signaling cascade and selectively function downstream of PKC isoforms. However, Bcl10 and Malt1 are not involved in Fc epsilon RI- or PKC-induced signaling events that control degranulation or leukotriene synthesis. Thus, the Bcl10/Malt1 complex specifically uncouples the pathway for cytokine production from degranulation events. This review will summarize our current knowledge of the regulation of Fc epsilon RI-induced NF-kappaB activation in mast cells and discuss potential implications for allergic inflammatory diseases. PMID- 17113920 TI - Subcortical control of dopamine neurons: the good, the bad and the unexpected. AB - The function of the phasic dopamine signal, seen in response to salient and rewarding stimuli, has been heavily debated. The reward prediction error hypothesis has been criticised for the suggestion that such a complex signal could be derived at short latencies, relying only on subcortical inputs. However, as more has been learnt about the nature of the subcortical inputs, we are led to challenge this criticism. Here we suggest that the subcortical inputs can indeed support complex calculations and that it would be unwise to underestimate their processing capabilities. Whilst our analysis cannot differentiate between the reward prediction error hypothesis and its opponents, it does suggest that the initial argument against a prediction error is incorrect. PMID- 17113921 TI - The role of sleep in the consolidation of route learning in humans: a behavioural study. AB - Considerable evidence support the role of sleep in learning and memory processes. In rodents, the relationships between sleep and memory consolidation have been extensively investigated by taking into account mainly spatial learning. On the contrary, in humans the relationship between sleep and spatial memory consolidation has so far been scarcely taken into account. Here, we investigated the importance of sleep in the consolidation of the spatial memory traces of a new route learned in a real-life unfamiliar environment. Fifty-one subjects followed a defined route in a neighbourhood they had never been to before. Then, they were tested in the laboratory in a sequence-recognition test requiring them to evaluate whether or not sequences of three views, taken along the route, represented a correct sequential order as seen while walking along the route. Participants were then assigned to one of three groups: the sleep group was retested after one night's sleep, the sleep-deprived group was retested after a night of sleep deprivation, and the day-control group was retested the same day after 8h of wakefulness. At retest, performance speed increased in all groups, whereas the accuracy in the sequence-recognition task was improved only in the sleep group: neither sleep deprivation nor the simple passage of time gave way to any performance improvement. These preliminary findings shed more light on the role of sleep in spatial memory consolidation by extending to humans the considerable evidence found in animals. PMID- 17113922 TI - Cerebellar interposed nucleus lesions suppress lymphocyte function in rats. AB - We previously reported that the cerebellar fastigial nucleus, output nucleus of the spinocerebellum, modulates lymphocyte function. To further explore the role of the cerebellum in neuroimmunomodulation, we here lesioned bilaterally the cerebellar interposed nuclei (IN) of rats with kainic acid (KA) injections. On days 8, 16 and 32 after IN lesions, lymphocyte percentage in peripheral white blood cells was examined. Furthermore, proliferation of lymphocytes from mesenteric lymph nodes induced by concanavalin A, sheep red blood cell-specific IgM antibody in the serum and cytotoxicity of natural killer cells from spleen against YAC-1 cells were measured by methyl-thiazole-tetrazolium assay, enzyme linked immunosorbent assay and flow cytometric assay, respectively. On days 8, 16 and 32 after KA injection in the IN, the lymphocyte percentage in the peripheral white blood cells was notably diminished with respect to control rats injected with saline in the IN. Concanavalin A-induced lymphocyte proliferation, serum sheep red blood cell-specific IgM antibody and natural killer cell toxicity of the IN-lesioned rats were significantly attenuated with respect to IN-saline control rats at all the post-lesion time points. The findings reveal that KA induced neuronal loss in the IN of both sides exerts an inhibitory effect on number and functions of T, B and natural killer lymphocytes, and indicate that the cerebellar IN participates in regulating immune function. Thus, the data suggest that the cerebellum may be an important brain area for neuroimmunomodulation, besides its well-known role in motor control. PMID- 17113923 TI - Forced swimming test increases superoxide anion positive cells and angiotensin II positive cells in the cerebrum and cerebellum of the rat. AB - Situations of stress are capable of inducing depression and oxidative stress in the brain. Previous reports have shown that angiotensin II (Ang II) induces the production of superoxide anion (O(2)(-)), and impairment of endothelial function in cerebral microvessels in vivo. Substances that reduce angiotensin functions may be important in the treatment of depression. These data suggest a role for both Ang II and O(2)(-) in depression; thus, the aim of this study was to determine the effect of forced swimming test (FST), a model of stress/depression, on the cellular expression of Ang II and O(2)(-) in the central nervous system. To induce stress/depression, rats were subjected to FST daily (30 min) for 15 days. Unstressed animals were used as controls. Motor activity was automatically analyzed daily before swimming. Cerebrum and cerebellum frozen sections were studied for O(2)(-) by a histochemical method and for Ang II producing cells by a polyclonal antibody. In the FST group, struggle time, total horizontal activity, ambulatory movements, and vertical movements, were significantly decreased when the data from the 1st and 15th day were compared. Food intake and body weight gain also decreased when unstressed and FST rats were compared at the 15th day. Increased number of cerebrum and cerebellum O(2)(-), and Ang II positive cells, were observed in FST rats. Significant correlation was found between O(2)(-) positive cells and Ang II positive cell in the cerebrum. These results suggest that stress/depression situations could be involved in the increase of Ang II and oxidative stress in the central nervous system, with possible implications in the depressive condition. PMID- 17113924 TI - The connexin 36 blockers quinine, quinidine and mefloquine inhibit cortical spreading depression in a rat neocortical slice model in vitro. AB - A protocol for inducing cortical spreading depression (SD) on rat neocortical slices in vitro, upon local application of calibrated approximately nl drops of KCl, 3M was used to elicit SD events, recorded at two different points on the slice. This in vitro model was validated by the inhibition of SD episodes by the NMDA antagonist MK-801 (20 microM), the reference SD blocker. Quinine, its stereoisomer quinidine, and mefloquine consistently inhibited the SD episodes. Quinine and quinidine, 100 and 200 microM reduced the duration, while mefloquine, 100 and 200 microM reduced the amplitude of SD events, all in a concentration dependent manner. These compounds have been reported to block gap junctions, specifically the neuronal connexin (Cx) 36, but they also exert other cellular effects. While further investigation is warranted to settle whether SD inhibition in vitro by quinine, quinidine and mefloquine reflects an involvement of neuronal Cx36 channels in SD generation/propagation, these results bear potential drug discovery relevance for the migraine with aura. PMID- 17113925 TI - Protective effect of rhubarb derivatives on amyloid beta (1-42) peptide-induced apoptosis in IMR-32 cells: a case of nutrigenomic. AB - Amyloid beta (1-42) peptide is considered responsible for the formation of senile plaques that accumulate in the brains of patients with Alzheimer's disease (AD). In the last years considerable attention has been focused on identifying natural food products, such as phytochemicals that prevent or almost retard the appearance of amyloid beta (1-42)-related neurotoxic effects. In this study, human neuroblastoma cells (IMR-32) was used as system model to evaluate the protective role of rhaponticin (3,3',5-trihydroxy-4'-methoxystilbene 3-O-d glucoside) a stilbene glucoside extracted from rhubarb roots (Rhei rhizoma) and rhapontigenin, its aglycone metabolite, against amyloid beta (1-42)-dependent toxicity. The obtained results show that rhapontigenin maintains significant cell viability in a dose-dependent manner and it exerts a protective effect on mitochondrial functionality, as evidenced by mitochondrial oxygen consumption experiments. A similar behaviour, but to a lesser extent, has been shown by rhaponticin. The protective mechanism mediated by the two stilbenes could be related to their effect on bcl-2 gene family expression. Bax, a pro-apoptotic gene, resulted down-regulated by the treatment with rhaponticin and rhapontigenin compared with the results obtained in the presence of amyloid beta (1-42) peptide. Conversely, bcl-2, an anti-apoptotic gene, highly down-regulated by amyloid beta (1-42) treatment, resulted expressed in the presence of stilbenes similarly to that shown by control cells. The obtained results support the hypothesis that amyloid beta (1-42)-induced neurotoxicity occurs via bax over expression, bcl-2 down-regulation, firstly indicating that rhaponticin and its aglycone moiety may alter this cell death pathway. Based on these studies, we suggest that rhaponticin and its main metabolite could be developed as agents for the management of AD. PMID- 17113926 TI - Cardiovascular responses produced by central injection of hydrogen peroxide in conscious rats. AB - Reactive oxygen species (ROS) have been shown to modulate neuronal synaptic transmission and may play a role on the autonomic control of the cardiovascular system. In this study we investigated the effects produced by hydrogen peroxide (H(2)O(2)) injected alone or combined with the anti-oxidant agent N-acetil-l cysteine (NAC) or catalase into the fourth brain ventricle (4th V) on mean arterial pressure and heart rate of conscious rats. Moreover the involvement of the autonomic nervous system on the cardiovascular responses to H(2)O(2) into the 4th V was also investigated. Male Holtzman rats (280-320 g) with a stainless steel cannula implanted into the 4th V and polyethylene cannulas inserted into the femoral artery and vein were used. Injections of H(2)O(2) (0.5, 1.0 and 1.5 micromol/0.2 microL, n=6) into the 4th V produced transient (for 10 min) dose dependent pressor responses. The 1.0 and 1.5 micromol doses of H(2)O(2) also produced a long lasting bradycardia (at least 24 h with the high dose of H(2)O(2)). Prior injection of N-acetyl-l-cysteine (250 nmol/1 microL/rat) into the 4th V blockade the pressor response and attenuated the bradycardic response to H(2)O(2) (1 micromol/0.5 microL/rat, n=7) into the 4th V. Intravenous (i.v.) atropine methyl bromide (1.0 mg/kg, n=11) abolished the bradycardia but did not affect the pressor response to H(2)O(2). Prazosin hydrochloride (1.0 mg/kg, n=6) i.v. abolished the pressor response but did not affect the bradycardia. The increase in the catalase activity (500 UEA/1 microL/rat injected into the 4th V) also abolished both, pressor and bradycardic responses to H(2)O(2). The results suggest that increased ROS availability into 4th V simultaneously activate sympathetic and parasympathetic outflow inducing pressor and bradycardic responses. PMID- 17113927 TI - Methylenetetrahydrofolate reductase gene polymorphisms are associated with ischemic and hemorrhagic stroke: Dual effect of MTHFR polymorphisms C677T and A1298C. AB - Hyperhomocysteinemia is an independent risk factor for ischemic stroke. The enzyme methylenetetrahydrofolate reductase (MTHFR) plays a critical role in modulating the levels of plasma homocysteine. Two polymorphisms in the MTHFR gene, C677T, A1298C result in reduced enzyme activity. The mechanisms of ischemic and hemorrhagic stroke are not well understood. Although controversial, previous studies have shown evidence of causality of both stroke subtypes in patients with methylenetetrahydrofolate reductase gene polymorphisms. Therefore, we examined whether the C677T and A1298C polymorphisms of MTHFR gene are genetic risk factors for both ischemic and hemorrhagic stroke in a Turkish Caucasian population. In a case-control study, 120 total unrelated stroke patients (92 ischemic stroke, 28 hemorrhagic stroke), and 259 healthy controls were genotyped for C677T and A1298C polymorphisms of the MTHFR gene using a PCR-RFLP based-method. The MTHFR 1298C allele (chi(2)=8.589; P=0.014), C1298C genotype (OR=2.544; P=0.004), and C677C/C1298C compound genotype (OR=3.020; P=0.001) were associated with overall stroke. The MTHFR 1298C allele (chi(2)=11.166; P=0.004), C1298C genotype (OR=2.950; P=0.001), and C677C/C1298C compound genotype (OR=3.463, P=0.0001) were strongly associated with ischemic stroke. Interestingly however, the MTHFR 677T allele (chi(2)=6.033; P=0.049), T677T genotype (OR=3.120; P=0.014), and T677T/A1298A compound genotype (OR=4.211; P=0.002) were associated with hemorrhagic stroke. In conclusion, the C677T and A1298C polymorphisms of the MTHFR gene are genetic risk factors for hamorrhagic and ischemic stroke respectively, independent of other atherothrombotic risk factors. PMID- 17113928 TI - Chemical stimulation of visceral afferents activates medullary neurones projecting to the central amygdala and periaqueductal grey. AB - Cholecystokinin (CCK) stimulates gastrointestinal vagal afferent neurones that signal visceral sensations. We wished to determine whether neurones of the nucleus of the solitary tract (NTS) or ventrolateral medulla (VLM) convey visceral afferent information to the central nucleus of the amygdala (CeA) or periaqueductal grey region (PAG), structures that play a key role in adaptive autonomic responses triggered by stress or fear. Male Sprague-Dawley rats received a unilateral microinjection of the tracer cholera toxin subunit B (CTB, 1%) into the CeA or PAG followed, 7 days later, by an injection of CCK (100 microg/kg, i.p.) or saline. Brains were processed for detection of Fos protein (Fos-IR) and CTB. CCK induced increased expression of Fos-IR in the NTS and the VLM, relative to control. When CTB was injected into the CeA, CTB-immunoreactive (CTB-IR) neurones were more numerous in the rostral NTS ipsilateral to the injection site, whereas they were homogeneously distributed throughout the VLM. Double-labelled neurones (Fos-IR+CTB-IR) were most numerous in the ipsilateral NTS and caudal VLM. The NTS contained the higher percentage of CTB-IR neurones activated by CCK. When CTB was injected into the PAG, CTB-IR neurones were more numerous in the ipsilateral NTS whereas they were distributed relatively evenly bilaterally in the rostral VLM. Double-labelled neurones were not differentially distributed along the rostrocaudal axis of the NTS but were more numerous in this structure when compared with the VLM. NTS and VLM neurones may convey visceral afferent information to the CeA and the PAG. PMID- 17113929 TI - Stimulation of mu and delta opioid receptors induces hyperalgesia while stimulation of kappa receptors induces antinociception in the hot plate test in the naked mole-rat (Heterocephalus glaber). AB - The antinociceptive effects of highly selective mu (DAMGO), delta (DPDPE) and kappa (U-50488 and U-69593) opioid agonists were evaluated following intraperitoneal (i.p.) administration in the naked mole-rat. A hot plate test set at 60 degrees C was used as a nociceptive test and the latency to the stamping of the right hind paw (response latency) was used as the end-point. DAMGO (5-10 mg/kg) and DPDPE (2.5-5 mg/kg) caused a naloxone-reversible significant decrease in the mean response latency. Subcutaneous injection of naloxonazine (20 mg/kg) 24h prior to the administration of DAMGO (5 mg/kg) also blocked the reduction in the response latency observed when DAMGO was injected alone. On the contrary, U 50488 (2.5-5 mg/kg) or U-69593 (0.08 or 0.1 mg/kg) caused a naloxone-reversible significant increase in the mean response latency. These results showed that activation of mu or delta receptors caused hyperalgesia, whereas activation of kappa receptors caused antinociception in the hot plate test in naked mole-rat. This suggests that mu and delta receptors modulate thermal pain in a different way than kappa receptors in the naked mole-rat. It is not possible at the moment to point out how they modulate thermal pain as little is known about the neuropharmacology of the naked mole-rat. PMID- 17113930 TI - Quantitative EEG analysis in post-traumatic anosmia. AB - Many objective and quantitative methods have been developed to create a procedure or a device to prove, describe and quantify olfactory deficit and anosmia, especially after a head trauma. Electrophysiological testing throughout olfactoelectroencephalography (olfactoEEG) is based on brain activity desynchronisation, and on the subsequent disappearance of alpha activity on the posterior regions after an olfactory stimulus. Yet traditional evaluation of EEG can be difficult, because of little or hardly detectable alpha activity on the posterior regions ('alpha rare'). The aim of this study was to evaluate the Olfactory Stop Reaction (OSR) by means of frequency band power calculation and subsequent topographical mapping in patients with post-traumatic anosmia, who presented 'alpha rare' EEG. Twenty-five consecutive patients, affected by anosmia caused by head trauma, were submitted to an EEG recording with olfactory stimulation. After signal processing and analysis, an Olfactory Stop Reaction was detected in 17 out of 25 patients; moreover, in these patients we detected a significant decrease in alpha band power in the occipital regions and an increase in theta band power on midline frontal and central regions after olfactory stimulation. In the remaining eight patients, no significant variation in band power was observed. In conclusion, an objective evaluation of the olfactory function with this method of automatic EEG signal analysis allows the limits given by psychophysical methods and traditional EEG to be overcome and attempts to fulfil the requirements for standardization of olfactory function evalution. PMID- 17113931 TI - Neural response to the visual familiarity of faces. AB - Recognizing personally familiar faces is the result of a spatially distributed process that involves visual perceptual areas and areas that play a role in other cognitive and social functions, such as the anterior paracingulate cortex, the precuneus and the amygdala [M.I. Gobbini, E. Leibenluft, N. Santiago, J.V. Haxby, Social and emotional attachment in the neural representation of faces, Neuroimage 22 (2004) 1628-1635; M.I. Gobbini, J.V. Haxby, Neural systems for recognition of familiar faces, Neuropsychologia, in press; E. Leibenluft, M.I. Gobbini, T. Harrison, J.V. Haxby, Mothers' neural activation in response to pictures of their, and other, children, Biol. Psychiatry 56 (2004) 225-232]. In order to isolate the role of visual familiarity in face recognition, we used fMRI to measure the response to faces characterized by experimentally induced visual familiarity that carried no biographical information or emotional content. The fMRI results showed a stronger response in the precuneus to the visually familiar faces consistent with studies that implicate this region in the retrieval of information from long-term memory and imagery. Moreover, this finding supports the hypothesis of a key role for the precuneus in the acquisition of familiarity with faces [H. Kosaka, M. Omori, T. Iidaka, T. Murata, T. Shimoyama, T. Okada, N. Sadato, Y. Yonekura, Y. Wada, Neural substrates participating in acquisition of facial familiarity: an fMRI study, Neuroimage 20 (2003) 1734-1742]. By contrast, the visually familiar faces evoked a weaker response in the fusiform gyrus, which may reflect the development of a sparser encoding or a reduced attentional load when processing stimuli that are familiar. The visually familiar faces also evoked a weaker response in the amygdala, supporting the proposed role of this structure in mediating the guarded attitude when meeting someone new. PMID- 17113932 TI - Relaxation strategies and enhancement of hypnotic susceptibility: EEG neurofeedback, progressive muscle relaxation and self-hypnosis. AB - Hypnosis has been shown to be efficacious in a range of clinical conditions, including the management of chronic pain. However, not all individuals are able to enter a hypnotic state, thereby limiting the clinical utility of this technique. We sought to determine whether hypnotic susceptibility could be increased using three methods thought to facilitate relaxation, with particular interest in an EEG neurofeedback protocol which elevated the theta to alpha ratio. This was compared with progressive muscle relaxation and self-hypnosis. Ten subjects with moderate levels of susceptibility (2-7/12) were randomly assigned to each condition and assessed for hypnotic susceptibility prior to and upon completion of 10 sessions of training. Hypnotic susceptibility increased post-training in all groups, providing further evidence that operant control over the theta/alpha ratio is possible, but contrary to our predictions, elevation of the theta/alpha ratio proved no more successful than the other interventions. Nonetheless, all three techniques successfully enhanced hypnotic susceptibility in over half of the participants (17/30), a similar incidence to that reported using other methods. As previously reported, the majority who were not susceptible to modification were at the lower levels of susceptibility, and the greater increases tended to occur in the more susceptible subjects. However, here enhancement was disclosed in some at low levels, and capability was found of reaching high levels, both features not typically reported. Further research is warranted. PMID- 17113933 TI - Localization of ZnT7 and zinc ions in mouse retina--immunohistochemistry and selenium autometallography. AB - Zinc transporter 7 (ZnT7, Slc30a7), a member of the Slc30 family, is involved in mobilizing zinc ions from the cytoplasm into the Golgi apparatus. In the present study, we examined the distribution and localization of ZnT7 and the labile zinc ions in the mouse retina using immunohistochemistry and in vivo zinc-selenium autometallography (ZnSe(AMG)). Our results showed that ZnT7 is abundantly expressed in the ganglion cells and pigment epithelial cells of the mouse retina. ZnT7 is also expressed in the amacrine cells and the layer of optic fibers of the mouse retina, but to a lesser extent. Weak staining of ZnT7 was detected in the inner plexiform layer, outer plexiform layer, and outer segment of the photoreceptors. However, ZnT7 was not detected in the outer nuclear layer and inner segment of the photoreceptors. A high level of labile zinc pool was detected in the pigment epithelial cells, the inner segment of the photoreceptors, and the marginal region of the inner nuclear layer. Less amount of labile zinc ions were detected in the ganglion cells of the retina. These observations strongly suggest that ZnT7 may play critical roles in retinal zinc homeostasis and that chelatable zinc pools may have multiple functions in the retina. PMID- 17113934 TI - Modulation of photic response by the metabotropic glutamate receptor agonist t ACPD. AB - Glutamate is the primary excitatory transmitter in the hypothalamus. It conveys photic information to the suprachiasmatic nucleus of the hypothalamus, thereby entraining the circadian clock to environmental light cycles. While ionotropic glutamate receptors have been implicated in the transduction of photic information in suprachiasmatic nucleus cells, there is evidence that metabotropic glutamate receptors play a significant modulatory role. We investigated the effects of the metabotropic glutamate agonist (+/-)-1-aminocyclopentane-trans-1,3 dicarboxylic acid (ACPD) on light-evoked phase responses in Syrian hamsters at three phase points: circadian time 6, a time when light has no effect on the circadian timing system; circadian time 13.5, when light evokes the maximum phase delay; circadian time 19, the maximum phase advance. We found that ACPD significantly increased the light-evoked phase shift at circadian time 13.5, and had no effect at other phase points tested. These data support a role for metabotropic glutamate receptors in the circadian photic signal transduction system. PMID- 17113935 TI - Vision of the hand prior to movement onset allows full motor adaptation to a multi-force environment. AB - In everyday life, because of unexpected mechanical perturbation applied to the hand or to the whole body, hand movements may become suddenly inaccurate. With prolonged exposure to the perturbation, trajectories slowly recover their normal accuracy, which is the mark of motor adaptation. However, full development of this adaptive process in complete darkness has been recently challenged in a multi-force environment. Here, we report on the effectiveness of static hand position information as specified through vision prior to movement onset on the adaptative changes, over trials, of pointing movements performed in a gravitoinertial force field. For this, subjects seated off-center on a platform rotating at constant velocity, were either confined to complete darkness (No Vision Session, NV) or provided with vision of the hand resting on the starting position prior to movement onset (Hand Vision Prior to Movement Session, HVPM). Overall, our results showed that adaptation to the centrifugal force was very rapid, and allowed subjects to demonstrate appropriate motor control as early as of the very first trials performed during the rotation period, even in the NV condition. They also showed that the integration by the Central Nervous System (CNS) of visual and proprioceptive information prior to the execution of a reaching movement allows subjects to reach full motor adaptation in a multi-force environment. Furthermore, our data confirm the existence of differentiated motor adaptive mechanisms for centrifugal and Coriolis forces. Adaptation to the former may fully develop on the basis of an a priori coding of the characteristics of the background force level even without visual information, while the latter needs visual cues about hand position prior to movement onset to take place. PMID- 17113936 TI - The effects of ethanol intake and withdrawal on penicillin-induced epileptiform activity in rats. AB - Previous experiments have shown that ethanol may have either pro-convulsive or anti-convulsive effects on epileptic activity in different experimental epilepsy models. In this study, the effect of low dose ethanol and its withdrawal on penicillin-induced epileptiform activity in rat was investigated. Eight groups of adult, male Wistar rats were studied: (1) control, (2) penicillin pretreated (500 units), (3) alpha-tocopherol (500 mg/kg, i.m.), (4) penicillin pretreated+alpha tocopherol, (5) ethanol-treated (3g/kg, per day, for 15 days, intragastrically)+penicillin, (6) ethanol-treated+penicillin+alpha-tocopherol, (7) ethanol withdrawal+penicillin, (8) ethanol withdrawal+penicillin+alpha tocopherol. Each animal group was composed of seven rats. The epileptiform activity was verified by electrocorticographic (ECoG) recordings. The epileptiform activity was induced by microinjection of penicillin into the left sensorimotor cortex. Administration of ethanol (3g/kg, per day, for 15 days, intragastrically) did not change either frequency or amplitude of penicillin induced epileptiform activity. The frequency and amplitude of epileptiform activity were evaluated 40 h after the last ethanol administration in withdrawal groups. There was no significant change in the mean frequency and amplitude of epileptiform activity compared with penicillin pretreated and ethanol-treated groups. The effective dose of alpha-tocopherol (500 mg/kg, i.m.) significantly decreased the mean frequency of epileptiform activity in the 60, 70, and 120 min after alpha-tocopherol injection in penicillin pretreated, ethanol-treated, ethanol withdrawal groups, respectively. However, alpha-tocopherol did not affect the amplitude of epileptiform activity in all groups. In conclusion, the present results indicate that low dose of ethanol does not have either anticonvulsive or proconvulsive effect on penicillin-induced epileptiform activity. alpha Tocopherol has anti-convulsive effect on penicillin-induced epileptiform activity in effective dose. PMID- 17113937 TI - Glial-derived neurotrophic factor (GDNF) prevents ethanol (EtOH) induced B92 glial cell death by both PI3K/AKT and MEK/ERK signaling pathways. AB - We investigated the neuroprotective effect of glial-derived neurotrophic factor (GDNF) upon alcohol-exposed B92 cultures, as well as the role of the cytoskeleton and mitogen-activated protein kinase (MAPK) pathways in this effect. Ethanol (EtOH) was added to cultures, either alone or in combination with 30 ng/ml GDNF. Exposure to EtOH (86 and 172 mM; 60 and 120 min) increased the frequency of apoptotic cells identified by nuclear DNA staining with 4,6-diamidino-2 phenylindole (DAPI). Cultures treated with GDNF showed a decrease in ethanol induced apoptosis. A jun N-terminal kinase (JNK) pathway is activated by EtOH and their pharmacological inhibition (by SP600125) neutralized ethanol-induced apoptosis, suggesting a role for JNK in EtOH neurotoxicity. Immunocytochemically detected phospho-JNK (p-JNK) showed an unusual filamental expression, and localized together with actin stress fibers. Examination of the cytoskeleton showed that EtOH depolymerized actin filaments, inducing p-JNK dissociation and translocation to the nucleus, which suggests that released p-JNK may contribute to glial cell death after EtOH exposure. Treatment with GDNF, in turn, may neutralize the ethanol-induced cell death pathway. Either a phosphatidylinositol 3-kinase (PI3K)/AKT pathway inhibitor (LY294002) or an inhibitor of the extracellular signal-regulated kinase (ERK) 1, 2 pathways (UO126) failed to neutralize GDNF protective effects. However, the simultaneous use of both inhibitors blocked the protective effect of GDNF, suggesting a role for both signaling cascades in the GDNF protection. These findings provide further insight into the mechanism involved in ethanol-induced apoptosis and the neurotrophic protection of glial cells. PMID- 17113938 TI - Drug resistance and hippocampal damage after delayed treatment of pilocarpine induced epilepsy in the rat. AB - Temporal lobe epilepsy (TLE) is the most common and pharmacoresistant form of epilepsy. Problems that cause pharmacoresistance may include delayed therapy due to late consultation, especially in developing countries. Our study aimed at unraveling consequences of delayed drug treatment using a rat model of TLE. Following pilocarpine-induced status epilepticus interrupted after 4h, rats were continuously videorecorded for onset and recurrence of spontaneous convulsive seizures. The animals were then treated for 50 days with carbamazepine (CBZ; first-line drug in TLE and effective also in rats), starting at seizure onset (27.22+/-3.38 days after status epilepticus) or 50 days later, and compared with epileptic untreated rats and non-epileptic CBZ-treated ones. Convulsive seizure frequency and duration, and hippocampal cell changes were evaluated. In particular, parvalbumin-containing hippocampal interneurons, astrocytes and microglia were characterized with immunohistochemistry and quantitative analyses. Prompt administration of CBZ suppressed seizures; delayed treatment only decreased frequency of convulsive seizures, which were also relatively prolonged. In hippocampal regions, histopathological damage, parvalbumin immunoreactivity loss, and glial activation were very marked after delayed treatment, and were reduced only slightly compared to untreated epilepsy, but enhanced compared to early treatment. The data on high frequency and duration of convulsive seizures in late-therapy rats indicate that delayed CBZ administration caused a high degree of drug resistance. This condition was subserved by severe damage in the hippocampus, presumably consequent to long-term seizure recurrence. Overall the data indicate that the paradigm of delayed treatment of limbic epilepsy could provide a model of drug-refractory TLE with hippocampal sclerosis. PMID- 17113939 TI - Translocation of cytochrome c during cerebellar degeneration in Lurcher and weaver mutant mice. AB - Cytochrome c translocation from the inner mitochondrial membrane into the cytosol is the initial step of the intrinsic apoptotic pathway. As no evidence was ever presented for cytochrome c translocation during cerebellar degeneration in Lurcher (Lc/+) and weaver (wv/wv) mutant mice, we searched for the presence of such a process in cerebellar homogenates of mutant and wild-type mice from postnatal day (P)1 to P56. Here we present the first documented time course of cytochrome c translocation spanning the entire period of neurodegeneration in both mutant types. We identified cytochrome c with Western blotting and monitored cell loss in the cerebellum with Calbindin D-28k immunohistochemistry, Nissl staining and morphometry. No cytochrome c translocation was ever detected in wild types at any age investigated. Translocated cytochrome c appeared between P13 and P21 in Lc/+ and between P5 and P6 in wv/wv. These two intervals precisely coincide with the respective periods of maximal neuronal death in the cerebellum. Secondary translocation was also observed at a later stage between P42 and P49 in Lc/+ and from P22 onwards in wv/wv. Since no substantial neuronal loss has ever been observed in Lc/+ and wv/wv mutants at these postnatal ages, the delayed translocation may correspond to cytochrome c of extraneuronal, presumably glial origin. Observations of an increased expression of glial fibrillary acidic protein and sustained remodeling of the astrocytic network in the cerebellum of both mutants, long after the cessation of neuronal death make this assumption rather plausible. PMID- 17113940 TI - Low-frequency subthalamic oscillations increase after deep brain stimulation in Parkinson's disease. AB - This work is the second of a series of papers in which we investigated the neurophysiological basis of deep brain stimulation (DBS) clinical efficacy using post-operative local field potential (LFP) recordings from DBS electrodes implanted in the subthalamic nucleus (STN) in patients with Parkinson's disease. We found that low-frequency (1-1.5Hz) oscillations in LFP recordings from the STN of patients with Parkinson's disease dramatically increase after DBS of the STN itself (log power change=0.93+/-0.62; Wilcoxon: p=0.0002, n=13), slowly decaying to baseline levels after turning DBS off. The DBS-induced increase of low frequency LFP oscillations is highly reproducible and appears only after the delivery of DBS for a time long enough to induce clinical improvement. This increase of low-frequency LFP oscillations could reflect stimulation-induced modulation of network activity or could represent changes of the electrochemical properties at the brain-electrode interface. PMID- 17113941 TI - Exercise intensity influences cell injury in rat hippocampal slices exposed to oxygen and glucose deprivation. AB - We evaluated the effects of two levels of daily forced exercise intensity (moderate and high) in the treadmill over cell susceptibility to oxygen and glucose deprivation (OGD) in hippocampal slices from Wistar rats. Moderate exercise decreased lactate dehydrogenase (LDH) release after OGD, while a significant increase in LDH release was observed in the high intensity group submitted to OGD. Our data corroborate the hypothesis that higher training intensity exacerbates brain damage, while a moderate intensity reduces the injury caused by in vitro ischemia. PMID- 17113942 TI - Opposite effects of a GABA(B) antagonist in two models of epileptic seizures in developing rats. AB - The action of a GABA(B) antagonist CGP 35348 and a GABA(B) agonist baclofen was studied in two models of epileptic seizures characterized by EEG spike-and-wave rhythm in freely moving immature rats. Rhythmic metrazol activity (RMA, model of human absences) was induced by low systemic dose of pentylenetetrazol (PTZ) in 18 and 25-day-old rats, epileptic after discharges (ADs, model of human myoclonic seizures) were elicited by electrical stimulation of sensorimotor cortex in rat pups 12, 18, and 25 days old. CGP 35348 (50, 100 and 200 mg/kg i.p.) suppressed RMA in both age groups in a dose-dependent manner. Simultaneously it increased the incidence of clonic seizures, potentiating thus an effect of PTZ. Baclofen (1, 3 and 6 mg/kg i.p.) augmented markedly RMA in 25-day-old rats. On the contrary, baclofen suppressed RMA in a part of 18-day-old animals. Incidence of seizures was not changed by baclofen in either age group. As ADs are concerned CGP 35348 (100 and 200 mg/kg i.p.) exhibited a proconvulsant action, baclofen (3, 6 or 12 mg/kg i.p.) was anticonvulsant, but again an irregularity of action was found in 18-day-old rats. The role of GABA(B)-mediated inhibition in epileptogenesis depends on the type of seizures and also on the stage of maturation. PMID- 17113943 TI - Recognition memory of newly learned faces. AB - We used event-related fMRI to study recognition memory of newly learned faces. Caucasian subjects memorized unfamiliar, neutral and happy South Korean faces and 4 days later performed a memory retrieval task in the MR scanner. We predicted that previously seen faces would be recognized faster and more accurately and would elicit stronger neural activation than novel faces. Consistent with our hypothesis, novel faces were recognized more slowly and less accurately than previously seen faces. We found activation in a distributed cortical network that included face-responsive regions in the visual cortex, parietal and prefrontal regions, and the hippocampus. Within all regions, correctly recognized, previously seen faces evoked stronger activation than novel faces. Additionally, in parietal and prefrontal cortices, stronger activation was observed during correct than incorrect trials. Finally, in the hippocampus, false alarms to happy faces elicited stronger responses than false alarms to neutral faces. Our findings suggest that face recognition memory is mediated by stimulus-specific representations stored in extrastriate regions; parietal and prefrontal regions where old and new items are classified; and the hippocampus where veridical memory traces are recovered. PMID- 17113944 TI - Exposure to high- and low-light conditions in an open-field test of anxiety increases c-Fos expression in specific subdivisions of the rat basolateral amygdaloid complex. AB - Anxiety states and anxiety-related behaviors appear to be regulated by a distributed and highly interconnected system of forebrain structures including the basolateral amygdaloid complex (basolateral amygdala). Despite a wealth of research examining the role of the basolateral amygdala in anxiety-related behaviors and anxiety states, the specific subdivisions of the basolateral amygdala that are involved in responses to anxiogenic stimuli have not been examined. In this study, we investigated the effects of exposure to a novel open field environment, with either low- or high-levels of illumination, on expression of the protein product of the immediate-early gene c-Fos in subdivisions of the rat basolateral amygdala. The subdivisions studied included the lateral, ventrolateral and ventromedial parts of the lateral amygdaloid nucleus, the anterior, posterior and ventral parts of the basolateral amygdaloid nucleus and the anterior and posterior part of the basomedial amygdaloid nucleus. Small increases in the number of c-Fos-immunoreactive cells were observed in several, but not all, of the subdivisions of the basolateral amygdala studied following exposure of rats to either the high- or low-light conditions, compared to home cage or handled control groups. Open-field exposure in both the high- and low light conditions resulted in a marked increase in c-Fos expression in the anterior part of the basolateral amygdaloid nucleus compared to either home cage or handled control groups. These findings point toward anatomical and functional heterogeneity within the basolateral amygdaloid complex and an important role of the anterior part of the basolateral amygdaloid nucleus in the neural mechanisms underlying physiological or behavioral responses to this anxiety-related stimulus. PMID- 17113945 TI - Involvement of anteroventral third ventricular AMPA/kainate receptors in both hyperosmotic and hypovolemic AVP secretion in conscious rats. AB - The area of the brain called the anteroventral third ventricular region (AV3V) includes three different subtypes of glutamate receptor, as well as neural circuits controlling fluid balance and cardiovascular and neuroendocrine functions. Although our previous data indicate the ability of AV3V N-methyl-d aspartate (NMDA) and metabotropic receptors to provoke vasopressin (AVP) releasing, pressor and hyperglycemic responses, the roles of non-NMDA receptors selective for alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid and kainate have not been elucidated to date. To address this question, the effects of intracerebral infusion with FWD or NBQX (specific agonist and antagonist for non-NMDA receptors, respectively) on plasma AVP, glucose, osmolality, electrolytes and cardiovascular parameters were examined in conscious rats in the absence or presence of an osmotic or volemic stimulus. When applied topically to AV3V structures such as the median preoptic nucleus, FWD augmented plasma AVP, osmolality, glucose and arterial pressure in a dose-associated fashion. All responses of the variables were abolished by pre-administering NBQX, which exerted no conspicuous effect on any variable except arterial pressure. It was revealed that NBQX administration in AV3V structures such as the median preoptic nucleus and the periventricular nucleus inhibited the rise of plasma AVP in response to intravenous infusion with hypertonic saline or removal of systemic blood through the femoral artery. Elevation of plasma osmolality and sodium evoked by osmotic load, and elevation of plasma osmolality, glucose and angiotensin II and decrease of arterial pressure caused by bleeding, were not significantly affected by NBQX treatment. These results suggest that AV3V non NMDA receptors, as well as NMDA receptors, may elicit AVP-releasing, pressor and hyperglycemic actions when stimulated in the basal state, and may facilitate AVP secretion under both hyperosmotic and hypovolemic conditions, without contributing to cardiovascular, blood glucose or other responses. PMID- 17113946 TI - Arginine vasopressin enhances periaqueductal gray synthesis and secretion of enkephalin and endorphin in the rat. AB - Previous study has proven that arginine vasopressin (AVP) enhances periaqueductal gray (PAG) secreting enkephalin and endorphin in vivo in the rat. Present work investigated that AVP effect on PAG secretion and synthesis of enkephalin and endorphin in vitro. Radioimmunoassy results showed that AVP increased leucine enkephalin (L-Ek), methionine-enkephalin (M-Ek), beta-endorphin (beta-Ep) rather than dynorphin A(1-13) (DynA(1-13)) concentrations in PAG slice culture liquid, and V(2) receptor antagonist: d(CH(2))(5)[D-Ile(2), Ile(4), Ala(9)-NH(2)]AVP decreased L-Ek, M-Ek and beta-Ep, not DynA(1-13) concentrations in PAG slice culture liquid in a dose-dependent manner, but V(1) receptor antagonist: d(CH(2))(5)Tyr(Me)AVP did not change these peptide concentrations in PAG slice culture liquid. RT-PCR data displayed that AVP enhanced proenkephalin and proopiomelanocortin (Pro-beta-Ep) rather than prodynorphin mRNA expressions in culture PAG tissues, and V(2) receptor antagonist weakened proenkephalin and proopiomelanocortin (Pro-beta-Ep), not prodynorphin mRNA expressions in culture PAG tissues, but V(1) receptor antagonist did not influence these mRNA expressions in culture PAG tissues. The data suggest that AVP enhances PAG synthesizing and secreting enkephalin and endorphin rather than dynorphin through V(2) receptors. PMID- 17113947 TI - Zolpidem, a selective GABA(A) receptor alpha1 subunit agonist, induces comparable Fos expression in oxytocinergic neurons of the hypothalamic paraventricular and accessory but not supraoptic nuclei in the rat. AB - Functional activation of oxytocinergic (OXY) cells in the hypothalamic paraventricular (PVN), supraoptic (SON), and accessory (ACC) nuclei was investigated in response to acute treatment with Zolpidem (a GABA(A) receptor agonist with selectivity for alpha(1) subunits) utilizing dual Fos/OXY immunohistochemistry. Zolpidem was administered intraperitoneally in dose 10 mg/kg of BW and 60 min later the animals were sacrificed by transcardial perfusion with fixative. The Fos/OXY co-labelings were analyzed on 40 microm thick serial coronal sections using computerized light microscopy. Zolpidem elicited a concordant Fos/OXY staining in all four PVN sub-areas investigated, including the anterior (15.71+/-2.35%), middle (14.52+/-2.53%), dorsal (13.34+/ 2.61%), and periventricular (18.21+/-4.75%) ones, however, had no significant stimulatory effect on OXY cells in the SON. In response to Zolpidem, statistically significant activations were also seen in certain groups of accessory structures including the circular nucleus (13.99+/-3.43%), small clusters of accessory neurons (10.55+/-1.94%), and the lateral hypothalamic perivascular nucleus (9.42+/-2.74%). Between the naive and vehicle controls, the dual Fos/OXY labelings did not elicit any significant differences. Our data provide insight into the topographic patterns of brain activity within the clusters of magnocellular OXY cells in the hypothalamus associated with stimulation of GABA(A) benzodiazepine receptors and for the first time illustrate the triggering contemporaneousness within the cells of the principal and accessory magnocellular nuclei in response to Zolpidem treatment. The present study provides a comparative background that may help in the further understanding of a possible extend of Zolpidem effect on the brain. PMID- 17113948 TI - The growth factor response in ischemic rat retina and superior colliculus after brimonidine pre-treatment. AB - The alpha-2-adrenergic receptor agonist brimonidine has been shown to increase survival of retinal ganglion cells following ischemic injury to the rat retina. Increased expression of growth factors has been suggested to be involved in this action. We investigated expressional changes of growth factors and their receptors following transient retinal ischemia induced by selective ligature of ophthalmic vessels in rats pre-treated with vehicle or 0.5% brimonidine. In addition, analysis of expression in retinal samples following unilateral administration of brimonidine to normal tissue was performed. Tissue samples of retina and superior colliculus were collected at time points between 6h and 14 days of retinal reperfusion. Analysis of mRNA levels of the ligands BDNF, NT3, CNTF, FGF1, FGF2, FGF9 and HGF; as well as the receptors TrkB, TrkC, p75(NTR), CNTFRalpha, FGFR1, FGFR3, FGFR4 and HGFR were performed using qRT-PCR. The cell specific markers Thy1 and GFAP were analysed. We report transiently increased retinal levels of BDNF, NT3, p75(NTR), FGFR1 and HGFR and decreased levels of FGF9, HGF, TrkB, TrkC, FGFR4 and Thy1 following ischemia. The decreases were counteracted by brimonidine. Brimonidine treatment gave an increase in BDNF, NT3 and CNTF levels compared to the vehicle treated group. In superior colliculus increased levels of growth factor mRNA were found. In conclusion, transient ischemia has a profound effect on gene expression in rat retina. Alterations can also be seen in the superior colliculus but are smaller. Brimonidine pre treatment attenuates an acute injury-induced response by decreasing the expression of several genes, among them p75(NTR). Brimonidine also causes a prolonged increase of several growth factors as well as receptors in retina and superior colliculus compared to the ischemic situation. The increased expression of several growth factors represents a coordinated growth factor system response that differs from the ischemia-induced changes and is likely part of the neuroprotective activity that is elicited by BMD pre-treatment. PMID- 17113949 TI - OFF response of bullfrog cones is shaped by terminal ionotropic GABA receptors. AB - We recently reported an ionotropic GABA receptor expressed at the bullfrog retinal cone terminal that is potentiated by the GABA(A) receptor antagonist bicuculline (BIC) and suppressed by the GABA(C) receptor antagonist imidazole-4 acetic acid (I4AA) . In this study, by using the patch clamp technique in current clamp mode, we show that activation of this GABA receptor causes voltage changes of cones, which are closely dependent on the membrane potential level in relation to the chloride equilibrium potential of the cells. Furthermore, the OFF overshoot of cone light responses is enhanced or diminished when this receptor is potentiated by BIC or suppressed by I4AA, suggesting the involvement of this GABA receptor in shaping OFF light responses of bullfrog cones. PMID- 17113950 TI - The occurrence of cone inclusions in the ageing human retina and their possible effect upon vision: an electron microscope study. AB - During normal ageing, photoreceptors of the human retina undergo various structural changes. We examined retinas from 33 donors (56 eyes; age span 13-94 years) by electron microscopy to see morphological changes in the cones with ageing. We show mitochondrial alterations and occurrence of electron-dense globules in the cone inner segments from the fifth decade of life. The globules are more prevalent in the macular cones than those in the mid-peripheral or nasal retinas (p<0.05) and absent in peripheral retinal cones and rods. They peak in the sixth decade and then decline in the seventh decade (p<0.05), from seventh to ninth decade, however, there was no significant change in their occurrence in the cones. We also show a type of inclusion, made up of bundled microtubules, which occur exclusively in the macular cones at the eighth decade of life. Evidence suggests that altered cone mitochondria with cristae remnants and dense matrix participate in globule formation in the ageing retina. Such mitochondrial changes may cause energy depletion, and bundling of microtubules (to form filamentous inclusions) could result in decreasing intracellular transport, in which case cones may die in the long run. These factors may be responsible for reported cone loss in the human retina with ageing. PMID- 17113951 TI - Activation of cerebellar nuclei comparing finger, foot and tongue movements as revealed by fMRI. AB - The aim of the present study was to compare possible activation of the interposed and dentate cerebellar nuclei during finger, foot and tongue movements using functional magnetic resonance imaging (fMRI). Nineteen healthy control subjects performed sequential finger and repetitive tongue and foot movement tasks. Thin slices (2.5mm) were acquired of the cerebellar region containing the cerebellar nuclei with high spatial resolution (matrix size 128 x 128 x 10) using a Siemens 1.5T Sonata system. Use of an eight channel head coil provided better signal-to noise-ratio compared to standard head coils. Only data of those 12 subjects were included in final statistical analysis, who showed significant activation of the cerebellar nuclei at least in one task. Cortical activations of the superior cerebellum were found in accordance to the known somatotopy of the human cerebellar cortex. Nuclear activations were most significant in the sequential finger movement task. Both interposed nuclei and ipsilateral dentate nucleus were activated. Dentate activation was present in the more caudal parts of both the dorsal and ventral nucleus. Activation overlapped with motor and non-motor domains of the dentate nucleus described by Dum and Strick [R.P. Dum, P.L. Strick, An unfolded map of the cerebellar dentate nucleus and its projections to the cerebral cortex, J. Neurophysiol. 89 (2003) 634-639] based on anatomical data in monkey. Tongue movement related activations were less extensive and overlapped with activations of caudal parts of the dentate nucleus in the finger movement task. No nuclear activation was seen following foot movements. The present findings show that both interposed and dentate nuclei are involved in sequential finger movements in humans. Interposed nucleus likely contributes to movement performance. Although no direct conclusions could be drawn based on the present data, different parts of the dentate nucleus may contribute to movement performance, planning and possible non-motor parts of the task. PMID- 17113952 TI - Purinergic nerves mediate the non-nitrergic relaxation of the human ileum in response to electrical field stimulation. AB - There has been no direct functional evidence for a purinergic innervation of the human intestinal muscle. In the present study, the relaxant effects of electrical field stimulation (1 or 10 Hz for 20s), ATP, and isoprenaline were studied in organ bath experiments on precontracted circular muscle strips of the human ileum. Non-adrenergic, non-cholinergic relaxations in response to electrical field stimulation in the presence of a nitric oxide synthase inhibitor were significantly reduced by the P(2) purinoceptor antagonists pyridoxal-phosphate-6 azophenyl-2',4'-disulphonic acid (PPADS; 50 microM) or suramin (100 microM). A combination of the two antagonists yielded an approximately 70% inhibition at 1 Hz. The relaxant effect of exogenous ATP, but not that of isoprenaline, was inhibited by PPADS+suramin. It is concluded that purinergic nerves (through P(2) purinoceptors) play a mediating role in the non-nitrergic relaxation in the human ileum. PMID- 17113953 TI - Decreased desychronisation during self-paced movements in frequency bands involving sensorimotor integration and motor functioning in Parkinson's disease. AB - This study examined sensorimotor integration and motor functioning in seven patients with Parkinson's disease (PD) who had mild symptoms, and seven age matched controls. Neuro-oscillations were recorded by high-density 128-channel electroencephalography (EEG). Participants were required to perform two tasks: simple tapping of the index finger and thumb and a complex Luria finger apposition task. Both tasks were performed unimanually and bimanually. There were no significant group differences in the task-related power (TRPow) within alpha 1 (mu1) or in beta 1 frequencies (beta1). In contrast, there were significant group differences in the alpha 2 (mu2) and beta 2 frequencies (beta2). Patients had less desychronisation than controls at the electrodes covering the central regions of the scalp. Alpha 2 and beta 2 frequencies have been associated with task-specific sensorimotor integration and motor function, respectively. This activity difference in patients with Parkinson's disease may be due to deficits in sensorimotor integration. PMID- 17113954 TI - GABA(B2) receptor subunit mRNA decreases in the thalamus of monoarthritic animals. AB - Many studies have implicated GABA(B) receptors in pain transmission mechanisms, especially in the spinal cord. In the thalamus, mRNA expression of the GABA(B(1b)) isoform was shown to be regulated in relay nuclei in response to chronic noxious input arising from experimental monoarthritis. GABA(B(1a)) and GABA(B2) mRNA expression was here determined by in situ hybridisation in the brain of control, 2, 4, 7 and 14 days monoarthritic rats, to evaluate whether this expression was regulated by chronic noxious input in thalamic nuclei. mRNA labelling was analysed quantitatively in the ventrobasal complex, posterior, central medial/central lateral and reticular thalamic nuclei; the thalamic visual relay and dentate gyrus were examined for control. No mRNA expression was detected for GABA(B(1a)) in control and monoarthritic animals. Similarly, GABA(B2) mRNA was not found in the reticular nucleus. However, GABA(B2) mRNA expression was observed in the ventrobasal complex, posterior and central medial/central lateral nuclei of control animals. A significant decrease of 42% at 2 days and 27% at 4 days of monoarthritis was observed in the ventrobasal complex contralaterally, when compared with controls, returning to basal levels at 7 days of monoarthritis. In the ipsilateral posterior nucleus, there was a significant decrease of 38% at 2 days of monoarthritis. No significant changes were observed in central medial/central lateral nuclei. The data suggest that GABA(B2) mRNA expression in the ventrobasal complex and posterior nucleus is regulated by noxious input and that GABA(B) receptors might play a role in the plasticity of these relay nuclei during chronic inflammatory pain. PMID- 17113955 TI - Functional changes in the activity of cerebellum and frontostriatal regions during externally and internally timed movement in Parkinson's disease. AB - We used fMRI to investigate the neurofunctional basis of externally and internally timed movements in Parkinson's disease (PD) patients. Ten PD patients whose medication had been withheld for at least 18h and 11 age- and sex-matched healthy controls were scanned while performing continuation paradigm with a visual metronome. Compared with the controls, PD patients displayed an intact capability to store and reproduce movement frequencies but with a significantly increased movement latencies. No differences in BOLD response were found in both groups when comparing the continuation with the preceding synchronization phase and viceversa, except for activity in visually related regions. Relative to healthy controls during the synchronization phase, PD patients exhibited an overall signal increase in the cerebellum and frontostriatal circuit (putamen, SMA and thalamus) activity together with specific brain areas (right inferior frontal gyrus and insula cortex) that are also implicated in primary timekeeper processes. By contrast, in the continuation phase the only neural network involved to a greater extent by the PD group was the cerebello-thalamic pathway. The lack of neurofunctional differences between the two timing phases suggests that rhythmic externally and internally guided movements engage similar neural networks in PD and matched healthy controls. Moreover, between-group comparison indicates that PD patients OFF medication may compensate for their basal ganglia cortical loop's dysfunction using different motor pathways involving cerebellum and basal ganglia relays during the two phases of rhythmic movement. PMID- 17113956 TI - Semantic priming effect during REM-sleep inertia in patients with narcolepsy. AB - Patients with narcolepsy-cataplexy (NC) present excessive daytime sleepiness (EDS), cataplexy and an altered architecture of nocturnal sleep, with frequent episodes of REM-sleep at sleep onset (SOREM-sleep). This altered organization of nocturnal sleep may be accompanied by some differences in the functioning of the cognitive processes involved in the access, organization and consolidation of information during sleep. This study attempts to ascertain whether the activation of semantic memory during REM-sleep, as measured using a technique of semantic priming (namely, the facilitation of the activation of strongly-related rather than weakly-related and, overall, unrelated pairs of prime-target words) is different in NC patients compared to normal subjects. A lexical decision task (LDT) was carried out twice in wakefulness (at 10a.m. and after a 24h interval) and twice in the period of sleep inertia following awakening from SOREM and 4th cycle REM-sleep on 12 NC patients and from 1st- and 4th-cycle REM-sleep on 12 matched controls. Reaction time (RT) to target words, taken as a measure of the semantic priming effect, proved to be longer (a) in NC patients than in control subjects; (b) in the period of REM-sleep inertia than in wakefulness; (c) in the first rather than the second session; and (d) for unrelated compared to weakly related and, overall, strongly-related prime-target pairs. RT in post-REM-sleep sessions was less impaired, compared to waking sessions, and less dependent on the associative strength of prime-target pairs in NC patients than in normal subjects. Finally, RT of NC patients, although longer than that of normal subjects in waking sessions, significantly improved in the second session, as a consequence of either the amount of exercise or the consolidation advantage provided by REM-sleep for the procedural components of the task. The whole picture suggests a greater effectiveness of the activation of semantic memory during (SO)REM-sleep in NC patients rather than in normal subjects, and overall for the organization of new and unexpected relationships (such as those between unrelated pairs) between items of information. PMID- 17113957 TI - Changes in pain behavior induced by formalin, substance P, glutamate and pro inflammatory cytokines in immobilization-induced stress mouse model. AB - In the present study, we examined the change of pain behaviors induced by formalin injected subcutaneously (s.c.) into the hind paw, or substance P (SP), glutamate, and pro-inflammatory cytokines (TNF-alpha, IL-1beta, and IFN-gamma) injected intrathecally (i.t.) in the mouse immobilization stress model. The mouse was restrained either once for 1h or five times for 5 days (once/day). In the formalin test, a single immobilization stress attenuated pain behaviors (licking, biting or scratching) in the second phase, while it had no effect on the pain behaviors revealed during the first phase. In addition, repeated immobilization stress attenuated pain behaviors revealed during the second phase but not in the first phase. A single as well as repeated immobilization stress decreased pain behaviors induced by substance P i.t. injection, but there were no significant changes in the glutamate test. In the pro-inflammatory cytokine pain model, a single immobilization stress decreased the pain behaviors induced by TNF-alpha, IL-1beta administered i.t. but not by IFN-gamma administered i.t. Moreover, a mouse applied with repeated immobilization stress did not show any changes in pain behaviors elicited by pro-inflammatory cytokines (TNF-alpha, IL-1beta and IFN-gamma) compared to the control group. These results suggest that a single and repeated immobilization stress differentially affects such nociceptive processing induced by formalin, SP, glutamate and pro-inflammatory cytokines in different manners. PMID- 17113958 TI - Co-modulation of acute ethanol-induced motor impairment by mouse cerebellar adenosinergic A1 and GABA(A) receptor systems. AB - We have previously demonstrated that cerebellar adenosine modulates ethanol ataxia. Using Rotorod method, we investigated the role of cerebellar GABA(A) receptors in the adenosinergic modulation of ethanol ataxia in mice. Direct cerebellar microinfusion of GABA(A) agonist, muscimol (2.5, 5 and 10 ng) and antagonist, bicuculline (50, 100 and 200 ng), via permanently implanted guide cannulas, produced a marked and dose-dependent accentuation and attenuation, respectively, of ethanol (2g/kg; IP) ataxia. The accentuation of ethanol ataxia by intracerebellar muscimol was through GABA(A) receptor because intracerebellar pretreatment with bicuculline virtually abolished muscimol effect. Intracerebellar microinfusion of adenosine A(1) agonist, N(6)-cyclohexyladenosine (CHA: 4 ng), and antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX: 100 ng) markedly accentuated and attenuated, respectively, ethanol ataxia consistent with our previously published data. Intracerebellar microinfusion of CHA (4 ng) or DPCPX (100 ng) markedly enhanced and reduced, respectively, muscimol (10 ng) induced accentuation of ethanol ataxia suggesting co-modulation of ethanol ataxia by cerebellar adenosinergic A(1) and GABA(A) receptors. Similarly, intracerebellar bicuculline (200 ng) pretreatment not only prevented CHA-induced accentuation of ethanol ataxia, but caused further decrease in ethanol ataxia. No change in the normal coordination was observed when microinfusion of the highest dose of muscimol, bicuculline, DPCPX or CHA alone or in combination was followed by saline injection instead of ethanol. The results of the present study suggest a functional similarity between GABA(A) and adenosine A(1) receptors even though both receptor types are known to couple to different signaling system and their location is on the opposite ends of the cerebellar granule cells, axons and axonal terminals (i.e., GABA(A) at the granule cells and adenosine A(1) on axons and axonal terminals of the granule cells) and act as co-modulators of ethanol ataxia. PMID- 17113959 TI - Olanzapine attenuates brain damage after focal cerebral ischemia in vivo. AB - Atypical antipsychotic drugs are widely used in the treatment of schizophrenia. These agents are discovered to have some additional beneficial effects beyond their effectiveness as antipsychotic drugs. Among these initially unexpected effects are their potential effects as mood stabilizers in bipolar disorder and their efficacy in improving long-term outcome in schizophrenia. These effects recently raised the question whether these drugs may also have some neuroprotective effect in the brain. To examine this matter, in this study we evaluated the neuroprotective effect of olanzapine after permanent focal cerebral ischemia. Anaesthetized male C57BL/6j mice were submitted to permanent thread occlusion of the middle cerebral artery (MCA). Olanzapine (0.1 and 1 mg/kg) or vehicle was applied intraperitoneally just after permanent ischemia. Twenty-four hours after permanent ischemia, brain injury was evaluated by triphenyltetrazolium chloride staining (TTC). Olanzapine (0.1 and 1 mg/kg) showed significant neuroprotection after permanent focal cerebral ischemia. PMID- 17113960 TI - Acute and chronic methylphenidate dose-response assessment on three adolescent male rat strains. AB - Methylphenidate (MPD), commonly known as Ritalin, is the most frequently prescribed drug to treat children and adults with attention deficit hyperactivity disorder (ADHD). Adolescence is a period of development involving numerous neuroplasticities throughout the central nervous system (CNS). Exposure to a psychostimulant such as MPD during this crucial period of neurodevelopment may cause transient or permanent changes in the CNS. Genetic variability may also influence these differences. Thus, the objective of the present study was to determine whether acute and chronic administration of MPD (0.6, 2.5, or 10.0mg/kg, i.p.) elicit effects among adolescent WKY, SHR, and SD rats and to compare whether there were strain differences. An automated, computerized, open field activity monitoring system was used to study the dose-response characteristics of acute and repeated MPD administration throughout the 11-day experimental protocol. Results showed that all three adolescent rat groups exhibited dose-response characteristics following acute and chronic MPD administration, as well as strain differences. These strain differences depended on the MPD dose and locomotor index. Chronic treatment of MPD in these animals did not elicit behavioral sensitization, a phenomenon described in adult rats that is characterized by the progressive augmentation of the locomotor response to repeated administration of the drug. These results suggest that the animal's age at time of drug treatment and strain/genetic variability play a crucial role in the acute and chronic effect of MPD and in the development of behavioral sensitization. PMID- 17113961 TI - Involvement of NMDA receptor mechanisms in the modulation of serotonin release in the lateral parabrachial nucleus in the rat. AB - Microdialysis was employed to investigate whether N-methyl-d-asparatate (NMDA) glutamate receptor mechanisms are involved in the modulation of serotonin (5 hydoxytryptamine, 5-HT) release in the region of the lateral parabrachial nucleus (LPBN) in freely moving rats. Perfusion of NMDA (10 and 50 microM) through the microdialysis probe significantly enhanced extracellular concentrations of 5-HT and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) in the LPBN area. Local perfusion of the NMDA antagonist dizocilpine (MK801, 10 and 50 microM) did not change the basal 5-HT and 5-HIAA levels in the LPBN area. MK801 (10 microM) administered together with NMDA antagonized the stimulant effect of NMDA (10 microM). The intake of 0.3M NaCl and water induced by subcutaneous injections of the diuretic furosemide (FURO, 10 mg/kg) and the angiotensin converting enzyme inhibitor captopril (CAP, 5 mg/kg) produced significant increases in the 5-HT and 5-HIAA concentrations in the LPBN area. The increased levels of 5-HT and 5-HIAA caused by the combined treatment with FURO and CAP were attenuated by perfusion of MK801 (10 microM). These results indicate the participation of NMDA receptors in the control of 5-HT release in the LPBN area. PMID- 17113962 TI - Electrophysiological correlates of visual affective priming. AB - The present study used event-related potentials (ERPs) to investigate the underlying neural mechanisms of visual affective priming. Eighteen young native English-speakers (6 males, 12 females) participated in the study. Two sets of 720 prime-target pairs (240 affectively congruent, 240 affectively incongruent, and 240 neutral) used either words or pictures as primes and only words as targets. ERPs were recorded from 64 scalp electrodes while participants pressed either "Happy" or "Sad" buttons to indicate target pleasantness. The response time (RT) results confirmed an affective priming effect, with faster responses to affectively congruent trials (659 ms) than affectively incongruent trials (690 ms). Affectively incongruent trials had larger and more negative N200 activation than those to neutral trials. Importantly, a delayed N400 for word prime-target pairs matched the RT results with larger negative amplitudes for incongruent than congruent pairs. This finding suggests that the N400 component is not only sensitive to semantic mismatches, but is also sensitive to affective mismatches for word prime-target pairs. PMID- 17113964 TI - Neutrophilic dermatoses. AB - Neutrophils may infiltrate all layers of the skin and consequently may cause different disorders, each with its own characteristic clinical and laboratory findings. We discuss how these disorders present and how they are diagnosed and treated. In addition, important associations with internal diseases are discussed to assist clinicians in evaluating for a concurrent illness. Because treatment of these disorders may often require systemic therapy, the potential short-term and long-term effects of commonly used medications are discussed. Finally, treatment of recalcitrant diseases, mostly by use of therapies published in the form of small case series or reports, is also included to guide clinicians in dealing with the more challenging cases. PMID- 17113965 TI - Relapsing polychondritis. AB - Relapsing polychondritis is a rare disease most commonly presenting as inflammation of the cartilage of the ears and nose. Auricular chondritis, with red ears resembling infectious cellulitis, is the most common initial finding. Antibodies to type II collagen in cartilage are found, and the earlobes are classically spared. Chronic disease may result in a flabby, droopy ear, cauliflower ear, or saddle nose deformity. Acute involvement of the tracheal cartilage may cause collapse of the airway with obstruction and pulmonary infections. Arthritis may be oligoarticular or polyarticular, most often involving the costochondral junctions. Other manifestations include audiovestibular damage; heart valve disease; and neurologic, ocular, and renal disease. Corticosteroids remain the major treatment. Other therapies include nonsteroidal anti-inflammatory drugs, dapsone, colchicine, azathioprine, methotrexate, cyclophosphamide, hydroxychloroquine, cyclosporine, and infliximab. PMID- 17113966 TI - Multicentric reticulohistiocytosis. AB - Multicentric reticulohistiocytosis is a rare granulomatous disease of unknown etiology, characterized by cutaneous nodules and destructive arthritis. Skin lesions can cause significant deformity, and approximately half of affected patients develop a severe disabling arthritis. The disease is often associated with malignancy; however, the paraneoplastic nature of multicentric reticulohistiocytosis is not established. The diagnosis is confirmed by the presence of oncocytic ("ground-glass") histiocytes and multinucleated giant cells on histopathology of the cutaneous nodules and the synovial membrane. PMID- 17113967 TI - Scleromyxedema. AB - Scleromyxedema is a rare cutaneous mucinous disease characterized by a generalized papular sclerodermoid eruption and systemic manifestations that can lead to significant morbidity and mortality. Although its etiology remains unknown, most theories focus on a pathogenic role by paraproteins; it must be noted, however, that nonparaprotein factors have been suggested to cause fibroblast proliferation and increased mucin production. Several treatment modalities including melphalan, cyclophosphamide, interferon alfa, and plasmapheresis have been suggested; however, further research is needed to prove treatment efficacy. PMID- 17113968 TI - Metabolic diseases: gout. AB - Gout is a disease of antiquity but is increasing once again in prevalence despite availability of reasonably effective treatments. This may be related to a combination of factors, including diet, obesity, and diuretic use. Allergic reactions, noncompliance, drug interactions, and sometimes inefficacy all limit the effective use of current hypouricemic agents. There are new treatments for gout on the horizon, including febuxostat, a nonpurine inhibitor of xanthine oxidase with a potentially better combination of efficacy and side effects than allopurinol. Diagnostic progress is being made in that ultrasound may offer a noninvasive means of diagnosing tophaceous deposits in and around joints. The increasing prevalence of gout means that dermatologists will see more cutaneous manifestations of gout, including tophi, draining sinus tracts, panniculitis, and dystrophic calcifications. PMID- 17113969 TI - Diagnosis, treatment, and prognosis of erythema migrans and Lyme arthritis. AB - Most patients with erythema migrans, the pathognomonic rash of Lyme disease, do not recall a deer tick bite. The rash is classically 5 to 68 cm of annular homogenous erythema (59%), central erythema (30%), central clearing (9%), or central purpura (2%). Serologic testing is not indicated for patients with erythema migrans, because initially, the result is usually negative. Successful treatment of a patient with erythema migrans can be accomplished with 20 days of oral doxycycline, amoxicillin, or cefuroxime axetil. Patients with Lyme arthritis usually present with a mildly painful swollen knee. Patients with Lyme arthritis have markedly positive serology and can usually be successfully treated with 28 days of oral doxycycline or amoxicillin. Some patients may have persistent effusion despite 4 to 8 weeks of antibiotics and may need synovectomy. Persistent effusion is not due to persistent infection. Antibiotic therapy for more than 8 weeks for patients with Lyme disease is not indicated. Chronic Lyme disease due to antibiotic resistant infection has not been demonstrated. PMID- 17113970 TI - Joint hypermobility and skin elasticity: the hereditary disorders of connective tissue. AB - The hereditary disorders of connective tissues (HDCTs) encompass a spectrum of conditions linked pathophysiologically by abnormalities of collagen, fibrillin, and matrix proteins. The clinical picture ranges from morbidity because of musculoskeletal, skin, ocular and visceral pathologies to mortality from acute vascular collapse. For many of the conditions, there is a considerable overlap in clinical features, although severity varies; appreciating the subtle differences in presentation is vital to the clinician in determining the diagnosis. Though conditions associated with severe vascular pathology are rare, other hereditary disorders of connective tissues such as the joint hypermobility syndrome and Stickler's disease are common and probably underrecognized. Abnormal skin elasticity and scaring, joint hypermobility, and chronic arthralgia are important clues that should trigger the clinician to search for underlying hereditary disorders of connective tissues. In this article, we discuss the spectrum of clinical findings, management, and genetic screening of the more common hereditary disorders of connective tissues, highlighting their diagnostic criteria and their differences. PMID- 17113971 TI - Human immunodeficiency virus infection and pregnancy. AB - Human immunodeficiency virus infection profoundly affects the medical community and is spreading rapidly in women of childbearing age worldwide. Transmission of HIV from mother to child can occur in utero, during labor, or after delivery through breast-feeding. Most of the infants are infected during delivery. We focus on the factors affecting the transmission of HIV, diagnostic and resistance tests, strategies to prevent mother-to-child transmission with special reference to mode of delivery, infant feeding, and use of antiretroviral therapy. The risk of infection for the infant can be decreased by reducing maternal viral load, by elective cesarean delivery, and by avoidance of breast-feeding. The efficacy of antiretroviral treatment should be balanced against the possibility of embryonic or fetal toxicity. The choice of therapy should be based on the woman's treatment history, the clinical status, and the available prognostic markers, which are related to the progression of disease in the mother and the risk of mother-to child transmission HIV transmission. PMID- 17113972 TI - Great chefs, dermatologists, and the black bag. PMID- 17113973 TI - From prurigo gestationis Besnier to atopic eruption of pregnancy: the confusing nosology of the less well-defined dermatoses of pregnancy has been largely clarified. PMID- 17113974 TI - Hulusi Behcet, MD: February 20, 1889 to March 8, 1948. PMID- 17113975 TI - Cosmetics preservation: sense and nonsense. PMID- 17113976 TI - Mechanisms of endothelin-1-induced MAP kinase activation in adrenal glomerulosa cells. AB - G protein-coupled receptors (GPCRs) such as angiotensin II, bradykinin and endothelin-1 (ET-1) are critically involved in the regulation of adrenal function, including aldosterone production from zona glomerulosa cells. Whereas, substantial data are available on the signaling mechanisms of ET-1 in cardiovascular tissues, such information in adrenal glomerulosa cells is lacking. Bovine adrenal glomerulosa (BAG) cells express receptors for endothelin-1 (ET-1) and their stimulation caused phosphorylation of Src (at Tyr416), proline-rich tyrosine kinase (Pyk2 at Tyr402), extracellularly regulated signal kinases (ERK1/2), and their dependent proteins, p90 ribosomal S6 kinase (RSK-1) and CREB. ET-1 elicited these responses predominantly through activation of a G(i)-linked cascade with a minor contribution from the G(q)/PKC pathway. Whereas, selective inhibition of EGF-R kinase with AG1478 caused complete inhibition of EGF-induced ERK/RSK-1/CREB activation, it caused only partial reduction (30-40%) of such ET-1 induced responses. Consistent with this, inhibition of matrix metalloproteinases (MMPs) with GM6001 reduced ERK1/2 activation by ET-1, consistent with partial involvement of the MMP-dependent EGF-R activation in this cascade. Activation of ERK/RSK-1/CREB by both ET-1 and EGF was abolished by inhibition of Src, indicating its central role in ET-1 signaling in BAG cells. Moreover, the signaling characteristics of ET-1 in cultured BAG cells closely resembled those observed in clonal adrenocortical H295R cells. The ET-1-induced proliferation of BAG and H295 R cells was much smaller than that induced by Ang II or FGF. These data demonstrate that ET-1 causes ERK/RSK-1/CREB phosphorylation predominantly through activation of G(i) and Src, with a minor contribution from MMP-dependent EGF-R transactivation. PMID- 17113977 TI - The role of estrogen in the initiation of breast cancer. AB - Estrogens are considered to play a major role in promoting the proliferation of both the normal and the neoplastic breast epithelium. Their role as breast carcinogens has long been suspected and recently confirmed by epidemiological studies. Three major mechanisms are postulated to be involved in their carcinogenic effects: stimulation of cellular proliferation through their receptor-mediated hormonal activity, direct genotoxic effects by increasing mutation rates through a cytochrome P450-mediated metabolic activation, and induction of aneuploidy. Recently it has been fully demonstrated that estrogens are carcinogenic in the human breast by testing in an experimental system the natural estrogen 17beta-estradiol (E(2)) by itself or its metabolites 2-hydroxy, 4-hydroxy, and 16-a-hydroxy-estradiol (2-OH-E(2), 4-OH-E(2), and 16-alpha-OH E(2)), respectively, by inducing neoplastic transformation of human breast epithelial cells (HBEC) MCF-10F in vitro to a degree at least similar to that induced by the chemical carcinogen benz(a)pyrene (BP). Neither Tamoxyfen (TAM) nor ICI-182,780 abrogated the transforming efficiency of estrogen or its metabolites. The E(2) induced expression of anchorage independent growth, loss of ductulogenesis in collagen, invasiveness in Matrigel, is associated with the loss of 9p11-13 and only invasive cells that exhibited a 4p15.3-16 deletion were tumorigenic. Tumors were poorly differentiated ER-alpha and progesterone receptor negative adenocarcinomas that expressed keratins, EMA and E-cadherin. The E(2) induced tumors and tumor-derived cell lines exhibited loss of chromosome 4, deletions in chromosomes 3p12.3-13, 8p11.1-21, 9p21-qter, and 18q, and gains in 1p, and 5q15-qter. The induction of complete transformation of the human breast epithelial cell MCF-10F in vitro confirms the carcinogenicity of E(2), supporting the concept that this hormone could act as an initiator of breast cancer in women. This model provides a unique system for understanding the genomic changes that intervene for leading normal cells to tumorigenesis and for testing the functional role of specific genomic events taking place during neoplastic transformation. PMID- 17113978 TI - Aromatase and breast cancer. AB - Several aromatase inhibitors and also new antiestrogens are now available for treating breast cancer. We have developed a model to compare the antitumor efficacy of these agents and to explore strategies for their optimal use. Results from the model have been predictive of clinical outcome. In this model, tumors are grown in ovariectomized, immunodeficient mice from MCF-7 human breast cancer cells transfected with the aromatase gene (MCF-7Ca). The possibility that blockade of estrogen action and estrogen synthesis may be synergistic was explored by treating mice with the aromatase inhibitor letrozole and the antiestrogen tamoxifen alone and in combination. The results indicated that letrozole alone was better than all other treatments. In addition, when tamoxifen treatment was no longer effective, tumor growth was significantly reduced in mice switched to letrozole treatment. However, tumors ultimately began to grow during continued treatment. To investigate the mechanisms by which tumors eventually adapt and grow during letrozole treatment, we determined the expression of signaling proteins in tumors during the course of letrozole treatment compared to the tumors of control mice. Tumors initially up-regulated the ER while responding to treatment, but subsequently receptor levels decreased in tumors unresponsive to letrozole. Also, Her-2 and adapter proteins (p-Shc and Grb-2) as well as all of the signaling proteins in the MAPK cascade (p-Raf, p-Mekl/2, and p-MAPK), but not in the Pl3/Akt pathway, were increased in tumors no longer responsive to letrozole. To investigate whether sensitivity to letrozole could be regained, cells were isolated from the letrozole resistant tumors (LTLT) and treated with inhibitors of the MAPKinase pathway (PD98059 and UO126). These compounds reduced MAPK activity and increased ER expression. EGFR/Her-2 inhibitors, gefitinib and AEE78S although not effective in the parental MCF-70a cells, restored the sensitivity of LTLT cells to letrozole. In xenografts, beginning treatment with letrozole and faslodex to down regulate the ER prevented increases in Her-2 and activation of MAPK and was highly effective in inhibiting tumor growth throughout 29 weeks of treatment. These results suggest that blocking both ER- and growth factor-mediated transcription may delay development of resistance and maintain growth inhibition of ER+ breast cancer. PMID- 17113979 TI - Vitamin D and cancer. AB - 1,25-dihydroxy Vitamin D [1,25-(OH)(2)D] exerts its effects via the vitamin D receptor (VDR) that belongs to the steroid/thyroid hormone receptor superfamily leading to gene regulation which results in various biological responses. Within the last two decades, the receptor has been shown to be present not only in classical target tissues such as bone, kidney and intestine but also in many other non-classical tissues. Besides the almost universal presence of VDRs, some cell types (e.g. keratinocytes, monocytes, bone, placenta) are capable of metabolizing 25-hydroxyvitamin D to 1,25(OH)(2)D by the enzyme 1alpha-hydroxylase (CYP27B1). The combined presence of 25(OH)D-1alpha-hydroxylase as well as the specific receptor in several tissues introduced the idea of a paracrine role for 1,25(OH)(2)D. Moreover, it has been demonstrated that 1,25(OH)(2)D can induce differentiation and inhibit proliferation of a wide variety of cell types. The molecular mechanisms behind this antiproliferative action is thoroughly explored but the whole picture is still difficult to understand. Important cell cycle regulators are involved such as cyclins, cyclin dependent kinases and their corresponding inhibitors as well as E2F transcription factors and accompanying pocket proteins. However the precise hierarchical structure of this wide diversity of actions of 1,25(OH)(2)D on genes influencing cell cycle progression is not firmly established nor do we understand which pathways are essential and which redundant. The antiproliferative action makes 1,25-(OH)(2)D and its analogs a possible therapeutic tool to treat hyperproliferative disorders, among which different types of cancer. This review focuses on the effects of 1,25(OH)(2)D and its analogs on cell proliferation, the results in in vivo experiments in Vitamin D deficient or resistant animals to cancer and the current epidemiological and intervention studies linking Vitamin D status or treatment and human cancer. PMID- 17113980 TI - Non-genomic steroid hormone effects: membrane or intracellular receptors? AB - A controversy regarding the identity of receptors that mediate nongenomic, transcription-independent cellular responses to steroids is presently attracting considerable scientific interest. While there is strong evidence for classic receptors belonging to the nuclear receptor superfamily to mediate nongenomic steroid effects in some cases, it does not exist for others. Nongenomic estrogen effects seem to predominantly involve classical estrogen receptors, both residing in cytoplasm and at the cell membrane. On the other hand, there is increasing evidence for the existence of nonrelated membrane receptors for estrogens, mediating CNS effects. Novel membrane receptors for other steroids have been recently cloned, with the demonstration of their biological relevance still largely pending. Recent findings on new and unexpected properties of classic receptors have partially deflected the interest from novel, nonclassic membrane receptors, which are being progressively identified at present. In addition, new findings pose challenges to some of the conclusions drawn from earlier experiments, and potential involvement of receptors and mechanisms of action need to be reconsidered. To know the nature of receptors involved will be key to beneficial medical translation of specific and targeted steroid responses. Differential pharmacological exploitation of different steroid receptors seems to become a tangible option. PMID- 17113981 TI - Biosynthesis, mode of action and functional significance of neurosteroids in the developing Purkinje cell. AB - New findings over the past decade have shown that the brain has the capability of forming steroids de novo from cholesterol, the so-called "neurosteroids". To understand neurosteroid action in the brain, data on the regio- and temporal specific synthesis of neurosteroids are needed. Recently, we have demonstrated that the Purkinje cell, a cerebellar neuron, is a major site for neurosteroid formation in a variety of vertebrates. This is the first demonstration of de novo neuronal neurosteroidogenesis in the brain. Since this discovery, organizing actions of neurosteroids are becoming clear by the studies on mammals using the Purkinje cell as an excellent cellular model. In mammals, the Purkinje cell actively synthesizes progesterone de novo from cholesterol during neonatal life, when cerebellar neuronal circuit formation occurs. The Purkinje cell may also produces estradiol in the neonate. Interestingly, both progesterone and estradiol promote dendritic growth, spinogenesis and synaptogenesis via each cognate nuclear receptor in the developing Purkinje cell. Such organizing actions may contribute to the formation of cerebellar neuronal circuit during neonatal life. This paper summarizes the advances made in our understanding of the biosynthesis, mode of action and functional significance of neurosteroids in the developing Purkinje cell. PMID- 17113982 TI - Metabolism of exogenous sex steroids and effect on brain functions with a focus on tibolone. AB - Around the menopause, changes in ovarian secretion of steroids result in changes in brain function: hot flushes and sweating later followed by changes in mood, libido and cognition. The relationship between sex steroids and brain functions are reviewed, with focus on hormonal treatments, in particular tibolone, on the postmenopausal brain and on associations between tissue levels and brain functions. Data on steroid levels in human brain are limited. Exogenous oestrogens alone or combined with progestagens reduce hot flushes and sweating, and may favourably affect anxiety, depression and mood. Testosterone alone or combined with E(2) improves libido and mood. Tibolone reduces hot flushes and sweating, and improves mood and libido, but does not stimulate endometrium or breast, like oestrogens. Tibolone is an ideal compound for studying steroid levels and metabolism in brain in view of its structural differences from endogenous steroids and its extensive metabolism required to express its endocrine effects. Brain levels of tibolone metabolites were measured in ovariectomized cynomolgus monkeys receiving tibolone for 36 days. Compared to serum, higher levels of the oestrogenic 3alpha/beta-hydroxytibolone and the androgenic/progestagenic Delta(4)-tibolone, and lower levels of sulphated metabolites are found in various brain regions. The high levels of oestrogenic metabolites in the hypothalamus explain hot flush reduction. Combined with the presence of Delta(4)-tibolone, the tibolone-induced increase in free testosterone through SHBG reduction explains androgenic effects of tibolone on mood and libido. The levels of tibolone metabolites in the monkey brain support tibolone's effects on brain functions. PMID- 17113983 TI - Prostate cancer risk in testosterone-treated men. AB - Men with classical androgen deficiency have reduced prostate volume and blood prostate-specific antigen (PSA) levels compared with their age peers. As it is plausible that androgen deficiency partially protects against prostate disease, and that restoring androgen exposure increases risk to that of eugonadal men of the same age, men using ART should have age-appropriate surveillance for prostate disease. This should comprise rectal examination and blood PSA measurement at regular intervals (determined by age and family history) according to the recommendations, permanently revisited, published by ISSAM, EAU, Endocrine Society.... Testosterone replacement therapy is now being prescribed more often for aging men, the same population in which prostate cancer incidence increases; it has been suggested that administration in men with unrecognised prostate cancer might promote the development of clinically significant disease. In hypogonadal men who were candidates for testosterone therapy, a 14% incidence of occult cancer was found. A percentage (15.2%) of prostate cancer has been found in the placebo group (with normal DRE and PSA) in the prostate cancer prevention study investigating the chemoprevention potential of finasteride. The hypothesis that high levels of circulating androgens is a risk factor for prostate cancer is supported by the dramatic regression, after castration, of tumour symptoms in men with advanced prostate cancer. However these effects, seen at a very late stage of cancer development, may not be relevant to reflect the effects of variations within a physiological range at an earlier stage. Data from all published prospective studies on circulating level of total and free testosterone do not support the hypothesis that high levels of circulating androgens are associated with an increased risk of prostate cancer. A study on a large prospective cohort of 10,049 men, contributes to the gathering evidence that the long standing "androgen hypothesis" of increasing risk with increasing androgen levels can be rejected, suggesting instead that high levels within the reference range of androgens, estrogens and adrenal androgens decrease aggressive prostate cancer risk. Indeed, high-grade prostate cancer has been associated with low plasma level of testosterone. Furthermore, pre-treatment total testosterone was an independent predictor of extraprostatic disease in patients with localized prostate cancer; as testosterone decreases, patients have an increased likelihood of non-organ confined disease and low serum testosterone levels are associated with positive surgical margins in radical retropubic prostatectomy. A clinical implication of these results concerns androgen supplementation which has become easier to administer with the advent of transdermal preparations (patch or gel) that achieve physiological testosterone serum levels without supra physiological escape levels. During the clinical development of a new testosterone patch in more than 200 primary or secondary hypogonadal patients, no prostate cancer was diagnosed. PMID- 17113984 TI - Noninvasive selective brain cooling by head and neck cooling is protective in severe traumatic brain injury. AB - Therapeutic hypothermia is a promising treatment for patients with severe traumatic brain injury (TBI). We present here the results of a study in which noninvasive selective brain cooling (SBC) was achieved using a head cap and neckband. Ninety patients with severe TBI were divided into a normothermia control group (n=45) and a SBC group (n=45), whose brain temperature was maintained at 33-35 degrees C for 3 days using a combination of head and neck cooling. At 24, 48 and 72h after injury, the mean intracranial pressure (ICP) values of the patients who underwent SBC were lower than those of the normothermia controls (19.14+/-2.33, 19.72+/-1.73 and 17.29+/-2.07 mmHg, versus 23.41+/-2.51, 20.97+/-1.86, and 20.13+/-1.87 mmHg, respectively, P<0.01). There was a significant difference in the neurological recovery of the two groups at the 6-month follow-up after TBI. Good neurological outcome (Glasgow Outcome Scale score of 4 to 5) rates 6 months after injury were 68.9% for the SBC group, and 46.7% for the control group (P<0.05). There were no complications resulting in severe sequelae. In conclusion, the noninvasive SBC described here is a safe method of administering therapeutic hypothermia, which can reduce ICP and improve prognosis without severe complications in patients with severe TBI. PMID- 17113985 TI - Metabotropic glutamate receptor antagonists and agonists: potential neuroprotectors in diffuse brain injury. AB - Our previous study has suggested that metabotropic glutamate receptors (mGluRs) were significantly involved in the secondary processes after diffuse brain injury (DBI) and that mGluRs antagonists or agonists may be used for the treatment of DBI. In the present study, the neuroprotective effects of antagonists or agonists of mGluRs on DBI were further investigated. Sprague-Dawly rats were randomized into the following six groups: (i) normal control; (ii) sham-operated control; (iii) DBI; (iv) DBI treated with normal saline (NS); (v) DBI treated with alpha methyl-4-carboxy-phenylglycine (MCPG); and (vi) DBI treated with (2S,1'R,2'R,3'R) 2-(2,3-dicarboxycyclopropyl)glycine (DCG-IV). Animals were injected intracerebroventricularly (icv) with 10 microL MCPG (100mmol/L), DCG-IV (10nmol/L) or the equivalent volume of normal saline 1 h after injury. The neurological severity score (NSS), brain water content and the number of damaged neurons were determined 6, 12, 24, 72 and 168 h after injury. In rats with DBI, it was found that the NSS was improved and the water content in the frontal cortex and the number of damaged neurons in the parietal cortex were significantly reduced following icv injection of either MCPG or DCG-IV. This suggests that icv injection of the mGluR group I antagonist MCPG or the mGluR group II agonist DCG-IV may exert neuroprotective effects in the early stage after DBI. PMID- 17113986 TI - Pituitary abscess. AB - Pituitary abscess is a rare disease, but one with potentially high mortality and morbidity. We present a 46-year-old man with progressive visual disturbance and general malaise for 1 year. Endocrine studies revealed hypopituitarism, and magnetic resonance imaging revealed a pituitary lesion with suprasellar extension. We attempted to excise the lesion using a transsphenoidal approach, but pus in the pituitary fossa was found at operation, and no tumour was identified. The culture yielded coagulase-negative Staphylococcus. Antibiotics were administered for 3 weeks, and the patient made a good postoperative recovery. He required life-long hormone replacement therapy. After one and a half years of follow-up, he was well and had no evidence of focal or systemic infection. We review the literature regarding pituitary abscess and discuss the appropriate treatment and possible pathological mechanism. PMID- 17113987 TI - Ganglion cyst of the cervical spine presenting with Brown-Sequard syndrome. AB - Ganglion cysts of the spine are uncommon. They occur mostly in the dorsolateral trunk and arise with the greatest frequency in the lumbar spine. However, they are rarely symptomatic. We report a rare case of a patient with a ganglion cyst of the lower cervical spine presenting with acute Brown-Sequard syndrome. The patient had no history of trauma. Magnetic resonance imaging of the cervical spine showed a cystic lesion connecting to the synovial joint C6-7 and compressing the posterior aspect of the spinal cord. The patient underwent emergent C6-7 laminectomy with total removal of the cyst. Neurological function recovered completely 4 months after operation. Ganglion cysts should be considered in the differential diagnosis of an extradural mass of the cervical spine. Magnetic resonance imaging provides a rapid and correct diagnosis, and laminectomy with removal of the cyst results in good neurological recovery. PMID- 17113988 TI - Cerebellopontine angle lipoma with extracranial extension. AB - Lipomas of the cerebellopontine angle (CPA) are rare. A recent literature review identified only 98 reported cases of CPA lipoma. We present here a case of CPA lipoma in a 28-year-old woman who was admitted to our hospital with hearing loss in her left ear. Computed tomography scan and magnetic resonance imaging revealed a CPA mass lesion with extracranial extension around the left internal carotiol artery. The patient was operated on in the sitting position via a right suboccipital craniectomy. The intracranial part of the mass was partially removed. Histopathological examination resulted in a diagnosis of lipoma. Surgical treatment of CPA lipomas is rarely indicated, and the aim of surgery must be decompression of neural structures. PMID- 17113989 TI - Brain abscess following intracerebral haemorrhage. AB - We report two cases of brain abscess, which developed at the site of an intracerebral haemorrhage (ICH) in a 75-year-old man and a 32-year-old-man. The patients recovered after surgical treatment and systemic antibiotic therapy. The route of infection could not be detected in either case. The literature contains only 13 reported cases of brain abscess as a complication of ICH. Although the interval from initial ICH to abscess formation ranged from 4 to 20 weeks, almost all patients had episodes of high fever, indicating the presence of systemic infection and bacterial seeding, 0-14 days after the onset of their ICH. Therefore, abscess formation appears to be caused by haematogenous seeding of infection in patients with ICH. Abscess formation should be considered when a patient deteriorates clinically with a febrile episode after an ICH. PMID- 17113990 TI - Granulomatous hypophysitis: presentation and MRI appearance. AB - Granulomatous hypophysitis (GrHy) is a relatively rare inflammatory disease compared with lymphocytic hypophysitis. Only a few cases with magnetic resonance imaging (MRI) findings have been reported to date. We describe the MRI findings for two patients with GrHy with unusual histories and clinical outcomes. PMID- 17113991 TI - Speedel staunches the renin cascade at its source. PMID- 17113992 TI - Targeting lipoxygenases with care. AB - Though fish oils possess cardio-protective, anti-inflammatory, and anti-cancer properties, their molecular and biochemical mechanism of action is lacking. In this issue of Chemistry & Biology, Tjonahen and colleagues identify a new metabolite of eicosapentanoic acid, resolvin E2, produced by 5-lipoxygenase. PMID- 17113993 TI - A new small-molecule Stat3 inhibitor. AB - In this issue of Chemistry & Biology, Schust et al. report the discovery of a small molecule (Stattic) that inhibits the binding of a high affinity phosphopeptide for the SH2 domain of Stat3. Stattic is a new tool for studying Stat3 signaling and demonstrates that the SH2 domain is not a dead target. PMID- 17113994 TI - Pseudouridine synthases. AB - Pseudouridine synthases are the enzymes responsible for the most abundant posttranscriptional modification of cellular RNAs. These enzymes catalyze the site-specific isomerization of uridine residues that are already part of an RNA chain, and appear to employ both sequence and structural information to achieve site specificity. Crystallographic analyses have demonstrated that all pseudouridine synthases share a common core fold and active site structure and that this core is modified by peripheral domains, accessory proteins, and guide RNAs to give rise to remarkable substrate versatility. PMID- 17113995 TI - Redesign of a central enzyme in alkaloid biosynthesis. AB - Plant alkaloids exhibit a diverse array of structures and pharmaceutical activities, though metabolic engineering efforts in these eukaryotic pathways have been limited. Strictosidine synthase (STR) is the first committed step in the biosynthesis of over two thousand terpene indole alkaloids. We describe a rational redesign of the STR binding pocket to selectively accommodate secologanin substrate analogs. The mutant is selective for a substrate that can be chemoselectively derivatized. Evidence that this substrate can be processed by later steps of the terpene indole alkaloid pathway is provided. The work demonstrates that the central enzyme of this alkaloid pathway can be redesigned and that the pathway can turn over the unnatural intermediate that is generated. Modulation of the substrate specificity of enzymes of this complex pathway is therefore likely to enable metabolic engineering efforts of these alkaloids. PMID- 17113996 TI - Insights into the synthesis of lipopolysaccharide and antibiotics through the structures of two retaining glycosyltransferases from family GT4. AB - Glycosyltransferases (GTs) catalyze the synthesis of the myriad glycoconjugates that are central to life. One of the largest families is GT4, which contains several enzymes of therapeutic significance, exemplified by WaaG and AviGT4. WaaG catalyses a key step in lipopolysaccharide synthesis, while AviGT4, produced by Streptomyces viridochromogenes, contributes to the synthesis of the antibiotic avilamycin A. Here we present the crystal structure of both WaaG and AviGT4. The two enzymes contain two "Rossmann-like" (beta/alpha/beta) domains characteristic of the GT-B fold. Both recognition of the donor substrate and the catalytic machinery is similar to other retaining GTs that display the GT-B fold. Structural information is discussed with respect to the evolution of GTs and the therapeutic significance of the two enzymes. PMID- 17113997 TI - Brasilicardin A, a natural immunosuppressant, targets amino Acid transport system L. AB - Lymphocytes in T cell activation require extracellular nutrients to provide energy for cellular proliferation and effector functions. Therefore, inhibitors of nutrient transporters are expected to be a new class of immunosuppressant. Here, we report that the molecular target of brasilicardin A (BraA), an immunosuppressive compound, is the amino acid transporter system L. BraA inhibited the cell-cycle progression of murine T cell lymphocyte CTLL-2 cells in G1 phase, and potently inhibited the uptake of amino acids that are substrates for amino acid transport system L. Moreover, BraA stimulated the GCN2 activation and, subsequently, the phosphorylation of eIF2alpha. These results suggest that the immunosuppressive activity of BraA is induced by amino acid deprivation via the inhibition of system L and that the amino acid transporter is a target for immunosuppressant. PMID- 17113998 TI - Biosynthesis of lovastatin analogs with a broadly specific acyltransferase. AB - The natural product lovastatin and its semisynthetic, more effective derivative, simvastatin, are important drugs for the treatment of hypercholesterolemia. Here, we report the biochemical characterization of a dedicated acyltransferase, LovD, encoded in the lovastatin biosynthetic pathway. We demonstrate that LovD has broad substrate specificity towards the acyl carrier, the acyl substrate, and the decalin acyl acceptor. LovD can efficiently catalyze the acyl transfer from coenzyme A thioesters or N-acetylcysteamine (SNAC) thioesters to monacolin J. When alpha-dimethylbutyryl-SNAC was used as the acyl donor, LovD was able to convert monacolin J and 6-hydroxyl-6-desmethylmonacolin J into simvastatin and huvastatin, respectively. Using the Escherichia coli LovD overexpression strain as a whole-cell biocatalyst, preparative amounts of simvastatin were synthesized in a single fermentation step. Our results demonstrate LovD is an attractive enzyme for engineered biosynthesis of pharmaceutically important cholesterol lowering drugs. PMID- 17113999 TI - Heterologous production of fosfomycin and identification of the minimal biosynthetic gene cluster. AB - Fosfomycin is a clinically utilized, highly effective antibiotic, which is active against methicillin- and vancomycin-resistant pathogens. Here we report the cloning and characterization of a complete fosfomycin biosynthetic cluster from Streptomyces fradiae and heterologous production of fosfomycin in S. lividans. Sequence analysis coupled with gene deletion and disruption revealed that the minimal cluster consists of fom1-4, fomA-D. A LuxR-type activator that was apparently required for heterologous fosfomycin production was also discovered approximately 13 kb away from the cluster and was named fomR. The genes fomE and fomF, previously thought to be involved in fosfomycin biosynthesis, were shown not to be essential by gene disruption. This work provides new insights into fosfomycin biosynthesis and opens the door for fosfomycin overproduction and creation of new analogs via biomolecular pathway engineering. PMID- 17114000 TI - Enzymatic generation of the antimetabolite gamma,gamma-dichloroaminobutyrate by NRPS and mononuclear iron halogenase action in a streptomycete. AB - Four adjacent open reading frames, cytC1-C4, were cloned from a cytotrienin producing strain of a Streptomyces sp. by using primers derived from the conserved region of a gene encoding a nonheme iron halogenase, CmaB, in coronamic acid biosynthesis. CytC1-3 were active after expression in Escherichia coli, and CytC4 was active after expression in Pseudomonas putida. CytC1, a relatively promiscuous adenylation enzyme, installs the aminoacyl moieties on the phosphopantetheinyl arm of the holo carrier protein CytC2. CytC3 is a nonheme iron halogenase that will generate both gamma-chloro- and gamma,gamma dichloroaminobutyryl-S-CytC2 from aminobutyryl-S-CytC2. CytC4, a thioesterase, hydrolytically releases the dichloroaminobutyrate, a known streptomycete antibiotic. Thus, this short four-protein pathway is likely the biosynthetic source of this amino acid antimetabolite. This four-enzyme system analogously converts the proS-methyl group of valine to the dichloromethyl product regio- and stereospecifically. PMID- 17114001 TI - Resolvin E2: identification and anti-inflammatory actions: pivotal role of human 5-lipoxygenase in resolvin E series biosynthesis. AB - The family of resolvins consists of omega-3 fatty acid-derived mediators, including E series resolvins generated from eicosapentaenoic acid (EPA), and carry potent anti-inflammatory properties. Here, we report the isolation, identification, and bioactions of resolvin E2 (RvE2), which is 5S,18-dihydroxy eicosapentaenoic acid. RvE2 stopped zymosan-induced polymorphonuclear (PMN) leukocyte infiltration and displayed potent anti-inflammatory properties in murine peritonitis. We also demonstrate that human recombinant 5-lipoxygenase generates RvE2 from a common precursor of E series resolvins, namely, 18 hydroxyeicosapentaenoate (18-HEPE). Furthermore, the initial 5-hydroperoxide intermediate was also converted to a 5(6)-epoxide intermediate in RvE1 formation. These results demonstrate that RvE2, together with RvE1, may contribute to the beneficial actions of omega-3 fatty acids in human diseases. Moreover, they indicate that the 5-lipoxygenase in human leukocytes is a pivotal enzyme that can produce both pro- and anti-inflammatory chemical mediators. PMID- 17114002 TI - Inhibition of Hsp90 with synthetic macrolactones: synthesis and structural and biological evaluation of ring and conformational analogs of radicicol. AB - A series of benzo-macrolactones of varying ring size and conformation has been prepared by chemical synthesis and evaluated by structural and biological techniques. Thus, 12- to 16-membered lactones were obtained by concise routes, involving ring-closing metathesis as a key step. In enzyme assays, the 13-, 15-, and 16-membered analogs are good inhibitors, suggesting that they can adopt the required conformation to fit in the ATP-binding site. This was confirmed by cocrystallization of 13-, 14-, and 15-membered lactones with the N-terminal domain of yeast Hsp90, showing that they bind similarly to the "natural" 14 membered radicicol. The most active compounds in the ATPase assays also showed the greatest growth-inhibitory potency in HCT116 human colon cancer cells and the established molecular signature of Hsp90 inhibition, i.e., depletion of client proteins with upregulation of Hsp70. PMID- 17114003 TI - A fluorescent broad-spectrum proteasome inhibitor for labeling proteasomes in vitro and in vivo. AB - The proteasome is an essential evolutionary conserved protease involved in many regulatory systems. Here, we describe the synthesis and characterization of the activity-based, fluorescent, and cell-permeable inhibitor Bodipy TMR-Ahx(3)L(3)VS (MV151), which specifically targets all active subunits of the proteasome and immunoproteasome in living cells, allowing for rapid and sensitive in-gel detection. The inhibition profile of a panel of commonly used proteasome inhibitors could be readily determined by MV151 labeling. Administration of MV151 to mice allowed for in vivo labeling of proteasomes, which correlated with inhibition of proteasomal degradation in the affected tissues. This probe can be used for many applications ranging from clinical profiling of proteasome activity, to biochemical analysis of subunit specificity of inhibitors, and to cell biological analysis of the proteasome function and dynamics in living cells. PMID- 17114004 TI - A monoselective sphingosine-1-phosphate receptor-1 agonist prevents allograft rejection in a stringent rat heart transplantation model. AB - FTY720 is an immunomodulator with demonstrated efficacy in a phase II trial of relapsing multiple sclerosis. FTY720-phosphate, the active metabolite generated upon phosphorylation in vivo, acts as a potent agonist on four of the five known sphingosine-1-phosphate (S1P(1)) receptors. AUY954, an aminocarboxylate analog of FTY720, is a low nanomolar, monoselective agonist of the S1P(1) receptor. Due to its selectivity and pharmacokinetic profile, AUY954 is an excellent pharmacological probe of S1P(1)-dependent phenomena. Oral administration of AUY954 induces a profound and reversible reduction of circulating lymphocytes and, in combination with RAD001 (Certican/Everolimus, an mTOR inhibitor), is capable of prolonging the survival of cardiac allografts in a stringent rat transplantation model. This demonstrates that a selective agonist of the S1P(1) receptor is sufficient to achieve efficacy in an animal model of transplantation. PMID- 17114005 TI - Stattic: a small-molecule inhibitor of STAT3 activation and dimerization. AB - Signal transducers and activators of transcription (STATs) are a family of latent cytoplasmic transcription factors that transmit signals from the cell membrane to the nucleus. One family member, STAT3, is constitutively activated by aberrant upstream tyrosine kinase activities in a broad spectrum of cancer cell lines and human tumors. Screening of chemical libraries led to the identification of Stattic, a nonpeptidic small molecule shown to selectively inhibit the function of the STAT3 SH2 domain regardless of the STAT3 activation state in vitro. Stattic selectively inhibits activation, dimerization, and nuclear translocation of STAT3 and increases the apoptotic rate of STAT3-dependent breast cancer cell lines. We propose Stattic as a tool for the inhibition of STAT3 in cell lines or animal tumor models displaying constitutive STAT3 activation. PMID- 17114006 TI - Who's going to do my operation? Expectations for the next generation of surgeons. PMID- 17114007 TI - 1423 pancreaticoduodenectomies for pancreatic cancer: A single-institution experience. AB - Pancreaticoduodenectomy (PD) with the possible addition of neoadjuvant or adjuvant therapy is the standard of care in the United States for adenocarcinoma originating in the pancreatic head, neck, and uncinate process. We reviewed 1423 patients who underwent a PD for a malignancy originating in the pancreas at our institution between 1970 and 2006. We examined 1175 PDs for ductal adenocarcinomas in greater detail. Eighteen different histological types of pancreatic cancer were identified; the most common diagnoses included ductal adenocarcinoma, neuroendocrine carcinoma, and IPMN with invasive cancer. Patients with ductal adenocarcinoma were analyzed in detail. The median age was 66 years, with patients in the present decade significantly older (68 years), on average, than patients in the three prior decades (e.g., 60 years in 1970, P = 0.02). The median tumor diameter was 3 cm; 42% of the resections had positive margins and 78% had positive lymph nodes. The perioperative morbidity was 38%. The median postoperative stay declined over time, from 16 days in the 1980s to 8 days in the 2000s (P < 0.001). The perioperative mortality declined from 30% in the 1970s to 1% in the 2000s (P < 0.001). The median survival for all patients with ductal adenocarcinoma was 18 months (1-year survival = 65 %, 2-year survival = 37%, 5 year survival = 18%). In a Cox proportional hazards model, pathological factors having a significant impact on survival included tumor diameter, resection margin status, lymph node status, and histologic grade. This is the largest single institution experience with PD for pancreatic cancer. Patients who have cancers with favorable pathological features have a statistically significant improved long-term survival. PMID- 17114008 TI - Pancreatic cancer in the general population: Improvements in survival over the last decade. AB - BACKGROUND: It is unknown whether the improved survival seen at high-volume centers has been translated to all patients with pancreatic cancer. OBJECTIVE: To use the Surveillance, Epidemiology, and End Results (SEER) database to evaluate population-based trends in surgical resection and survival. METHODS: All patients diagnosed with pancreatic cancer from 1988-1999 were identified. The survival and proportion of patients undergoing surgical resection were compared for each of three equal time periods. RESULTS: There were 24,016 patients with pancreatic cancer. 19,533 had stage data available. 9% had localized, 29% had regional, and 62% had distant disease. Resection rates increased for patients with localized and regional disease over the three time periods. Survival increased for patients with regional and distant disease. For regional pancreatic cancer patients, 2 year survival increased from 9.5% to 13.5% (p < 0.0001) and from 21.5% to 28.9% following surgical resection (p = 0.002). For resected local/regional pancreatic cancer, the year of diagnosis was and independent predictor of improved survival (p = 0.0001). CONCLUSIONS: SEER patients with regional and distant pancreatic cancer have improved survival over the past decade in both unadjusted and adjusted models. The improvement is most striking for patients with regional disease and reflects increased resection rates and improved resection techniques over time. PMID- 17114009 TI - Exocrine function following the whipple operation as assessed by stool elastase. AB - What impact does pancreaticoduodenectomy (PD) have on exocrine function? Does the pancreatic anastomosis remain patent? When stool elastase became available for testing in November 2001, we began preoperative assessment and then increasingly employed postoperative measurements. From December 2001 until March 2006, 182 patients underwent PD by the same surgeon. Preoperative stool elastase was measured in 138 (76%) patients and was repeated postoperatively at 3 +/- 1 month, 12 +/- 2 months, and 24 +/- 3 months. At the same time periods, an abdominal CT scan was used to assess patency of the pancreatic anastomosis as implied by pancreatic duct dilation in the remnant (dilation = duct >3 mm or, if duct dilated preoperatively, then duct that failed to decrease in size). All cases were reconstructed with duct-to-mucosa pancreaticojejunostomy. Stool elastase was expressed as normal (>200 microg/gram stool), moderately reduced (100-200 microg/gram), or severely reduced (<100 microg/gram). Preoperative stool elastase values were "normal" in 78% (pancreatic cancer 32% normal vs. all other groups >78%; P < or = 0.001). As compared with preoperative values, the percent of cases with reduced elastase levels at 3 months, 1 year, and 2 years postoperatively was 48%, 73%, and 50%, respectively. The CT scans at the time of the 69 stool elastase measurements after PD showed pancreatic duct dilation in the pancreatic remnant in 9 of 69 (9%) stools but was not more frequent in the group with decreased elastase. Based on cases elastase, one third of patients about to have PD will have exocrine insufficiency, an observation most common among the patients with pancreatic cancer (68%). Stool elastase levels are further depressed in the majority of cases after PD from parenchymal loss because we could not implicate an occluded pancreatic anastomosis. These results suggest that, after PD, exocrine supplementation should be given to all patients with pancreatic cancer, especially those with impending adjuvant therapy. To further improve the long-term results after PD, each surgeon should assess the effect of their own type of pancreaticoenteric technique on exocrine function. PMID- 17114010 TI - Pancreatic regeneration in chronic pancreatitis requires activation of the notch signaling pathway. AB - Chronic pancreatitis as an inflammatory process characterized by morphological changes, pancreatic dysfunction, and pain. During pancreatic injury and repair the Notch signaling pathway is reinstated. The current study analyzed this pathway in chronic pancreatitis and characterized its influence on fibrogenesis. Real-time quantitative PCR and immunohistochemistry were used for expression studies. Notch activation was determined by a specific luciferase-HES-1-reporter gene constructs. Cells were stimulated with alcohol, glucose, bile acids, and steroids. Notch-2, -3, and -4 mRNA, were overexpressed in chronic pancreatitis specimens. The ligands Jagged-1, -2, and Delta-1 were highly overexpressed. Jagged-1 and Notch receptors were observed in nerves, regenerating exocrine cells, and endocrine cells. Delta staining was present in ductal but not in acinus cells and not in nerves. Activation of Notch signaling was detectable upon cell stimulation with glucose, steroids, and bile acids. High glucose levels were further associated with increased collagen-I production. The Notch pathway is reactivated during chronic pancreatitis. Among the stimuli activating the Notch pathway are steroids, high glucose levels, and bile acids. These findings suggest a possible role of the Notch pathway during pancreatic regeneration since Jagged 1 inhibits inducible collagen-1 production, suggesting a new mechanism of tissue repair in this disease. PMID- 17114011 TI - Hospital readmission after pancreaticoduodenectomy. AB - Data exist on the morbidity and mortality of patients undergoing pancreaticoduodenectomy (PD), but there are few reports about hospital readmissions after this procedure. Our aim was to evaluate the number of and reasons for readmission after PD and the factors influencing readmission. We reviewed the initial hospitalization and readmissions for 1643 patients undergoing PD compared patients requiring readmission to patients that did not require readmission. Twenty-six percent of patients were readmitted a total of 678 times after PD. Patients readmitted were younger than those not readmitted (61.8 versus 64.6 years, P < 0.0001). Vessel resection, abscess formation, wound infection, postoperative percutaneous biliary stents, estimated blood loss >1000 ml, and age < or =65 years were independently associated with readmission. The length of stay for all patients decreased over time, from 10.5 days in 1996 to 7 days in 2003. The percentage of patients being readmitted also decreased from 33% in 1996 to 20% (P = 0.004) in 2003. The readmission rate after PD was 26%. Younger age, blood loss, postoperative complications, and vessel resection were independent risk factors for readmission. The early hospital readmission rate has not increased in association with a decreased LOS, supporting the idea that reduction in LOS did not lead to increased readmission rates. PMID- 17114012 TI - Lentivirus-mediated RNA interference of HMGA1 promotes chemosensitivity to gemcitabine in pancreatic adenocarcinoma. AB - The high mobility group A1 (HMGA1) proteins are overexpressed in pancreatic cancers. They are architectural nuclear proteins, which regulate expression of multiple genes implicated in the malignant phenotype. In this study, we hypothesized that HMG A1 silencing will promote chemosensitivity in pancreatic adenocarcinoma. We studied highly malignant pancreatic adenocarcinoma cell lines (MiaPaCa2 and PANC1). Lentiviral short-hairpin RNA (shHMGA1) expression vectors targeting HMGA1 were used for generation of lentiviral particles. Stable transfectants were developed after lentiviral transduction. Nuclear expression of HMGA1 was assayed using Western blot analysis. Chemosensitivity to gemcitabine was determined by IC50 analysis. Caspase activity was quantitated using fluorometric caspase profiling. Apoptosis was assessed by flow cytometric analysis. Lentivirus-mediated RNA interference resulted in 90% silencing of HMGA1 expression in each of MiaPaCa2 and PANC1 cell lines. HMGA1 silencing enhanced chemosensitivity to gemcitabine with an approximately 50% reduction in IC50 in each cell line. Lentivirus-mediated HMGA1 silencing promoted the activation of caspases 3, 2, 9, and 8, on exposure to gemcitabine. HMGA1 silencing resulted in reduction in Akt kinase activity. Lentivirus-mediated RNA interference of HMGA1 promoted chemosensitivity to gemcitabine in pancreatic adenocarcinoma. HMGA1 may represent a novel therapeutic target in pancreatic cancer. PMID- 17114013 TI - Postoperative pancreatic fistulas are not equivalent after proximal, distal, and central pancreatectomy. AB - It is uncertain whether postoperative pancreatic fistulas after distal and central pancreatectomies behave similarly to those after pancreaticoduodenectomy. To date, this concept has not been validated either clinically or economically. Overall, 256 consecutive pancreatic resections from October 2001 to February 2006 (184 pancreaticoduodenectomies, 66 distal pancreatectomies, and 6 central pancreatectomies) were evaluated according to the International Study Group of Pancreatic Fistula classification scheme. Pancreatic fistula was defined as any measurable drainage on or after postoperative day 3, with amylase content greater than three times the normal serum value. Outcomes were divided into four grades: (1) no fistula, (2) grade A: biochemical fistula without clinical sequelae, (3) grade B: fistula requiring any therapeutic intervention, or (4) grade C: fistula with severe clinical sequelae. Grades B and C are considered clinically relevant fistulas based on worsening morbidity, increased length of stay, frequent hospital readmission, and increased costs/resource utilization. Clinical and economic outcomes were compared-grade for grade-across the three resection types. Fistulas of any extent (Grades A-C) occurred in one third of all patients; two thirds had no fistula. Overall, there were 16 readmissions (6%), six reoperations (2%), and no deaths attributable to pancreatic fistula. Outcomes between no fistula and grade A patients were identical across resection types, though grade A fistula was more common in distal pancreatectomy. For each resection type, length of stay and costs progressively increased with grades B and C. However, the negative impact of these clinically relevant fistulas varied between resection types. Rates for intensive care unit admission and rehabilitation placement were higher among pancreaticoduodenectomy patients. Total parenteral nutrition and antibiotic use were similar, but percutaneous drainage was used more often for distal pancreatectomy. Grade B fistula was more severe after distal pancreatectomy, as indicated by increased length of stay, readmissions, and total cost. Although reoperation rates for grade C fistulas were equivalent, intervals to reoperation were substantially longer after distal and central pancreatectomies. When classified according to International Study Group of Pancreatic Fistula criteria, clinically relevant pancreatic fistulas behaved differently depending on type of pancreatectomy. This translates into variable severity that guides management decisions, which ultimately dictate clinical outcomes and economic impact. PMID- 17114014 TI - Does pancreatic duct stenting decrease the rate of pancreatic fistula following pancreaticoduodenectomy? Results of a prospective randomized trial. AB - Pancreatic duct stenting remains an attractive strategy to reduce the incidence of pancreatic fistulas following pancreaticoduodenectomy (PD) with encouraging results in both retrospective and prospective studies. We performed a prospective randomized trial to test the hypothesis that internal pancreatic duct stenting reduces the development of pancreatic fistulas following PD. Two hundred thirty eight patients were randomized to either receive a pancreatic stent (S) or no stent (NS), and stratified according to the texture of the pancreatic remnant (soft/normal versus hard). Four patients were excluded from the study; in three instances due to a pancreatic duct that was too small to cannulate and in the other instance because a total pancreatectomy was performed. Patients who randomized to the S group had a 6-cm-long segment of a plastic pediatric feeding tube used to stent the pancreaticojejunostomy anastomosis. In patients with a soft pancreas, 57 randomized to the S group and 56 randomized to the NS group. In patients with a hard pancreas, 58 randomized to the S group and 63 randomized to the NS group. The S and NS groups for the entire study population, as well as for the subgroup of high-risk patients with soft pancreata, were similar as regard to demographics, past medical history, preoperative symptoms, preoperative procedures, and intraoperative data. The pancreatic fistula rate for the entire study population was 9.4%. The fistula rates in the S and NS subgroups with hard pancreata were similar, at 1.7% and 4.8% (P = 0.4), respectively. The fistula rates in the S and NS subgroups with soft pancreata were also similar, at 21.1% and 10.7% (P = 0.1), respectively. A nonstatistically significant increase in the pancreatic fistula rate in the S group persisted after adjusting for the operating surgeon and technical details of the operation (e.g., anastomotic technique, anastomotic orientation, pancreatic duct size, and number of intra abdominal drains placed). In patients with soft pancreata, 63% percent of the pancreatic fistulas in stented patients required adjustment to the clinical pathway (including two deaths), compared to 47% of the pancreatic fistulas in patients in the NS group (P = 0.3). Internal pancreatic duct stenting does not decrease the frequency or the severity of postoperative pancreatic fistulas. PMID- 17114015 TI - Treatment with gemcitabine and TRA-8 anti-death receptor-5 mAb reduces pancreatic adenocarcinoma cell viability in vitro and growth in vivo. AB - Gemcitabine is a first line agent for pancreatic cancer, but yields minimal survival benefit. This study evaluated in vitro and in vivo effects of a monoclonal antibody (TRA-8) to human death receptor 5, combined with gemcitabine, using two human pancreatic cancer cell lines, S2VP10 and MIA PaCa-2. A subcutaneous model of pancreatic cancer was employed to test in vivo efficacy. S2VP10 and MIA PaCa-2 cells were treated with varying doses of gemcitabine and TRA-8. Cell viability and apoptosis were determined with an adenosine triphosphate assay and annexin V staining, respectively. Mitochondrial membrane destabilization was evaluated with fluorescence-activated cell sorting analysis of JC-1 stained cells. Caspase activation was evaluated by Western blot analysis. MIA PaCa-2 subcutaneous xenografts in athymic nude mice were evaluated for response to treatment with 200 mug of TRA-8 (intraperitoneal on days 9, 13, 16, 20, 23, and 27 postimplant) and 120 mg/kg gemcitabine (I.P. on days 10, 17, and 24). Tumor growth was measured with calipers. MIA PaCa-2 and S2VP10 cells receiving combination treatment with TRA-8 and gemcitabine demonstrated enhanced cytotoxicity, annexin V staining, and mitochondrial destabilization compared to either agent alone. Combination treatment produced enhanced caspase-3 and -8 activation in both cell lines compared with either agent alone. In vivo studies demonstrated mean subcutaneous tumor surface area (produce of two largest diameters) doubling times of 38 days untreated, 32 days gemcitabine, 49 days TRA 8, and 64 days combination treatment. TRA-8 is an apoptosis-inducing agonistic monoclonal antibody that produced synergistic cytotoxicity in combination with gemcitabine in vitro through enhanced caspase activation. These findings, with substantial inhibition of tumor growth in a mouse pancreatic cancer xenograft model receiving combination therapy, are encouraging for anti-death receptor therapy in the treatment of pancreatic cancer. PMID- 17114016 TI - The effects of neoadjuvant chemoradiation on pTNM staging and its prognostic significance in esophageal cancer. AB - For esophageal cancer, it is not clear if pathologic TNM staging after chemoradiation and resection will have the same prognostic significance compared with patients who undergo resection only. From 1995 to 2004, prospectively collected data from 279 patients with intrathoracic squamous cell cancers were analyzed. Patients were given chemoradiation either as part of a randomized trial comparing neoadjuvant chemoradiation with surgical resection alone, or because of advanced disease at presentation. One hundred seventy patients had surgical resection only (surgery), and 109 had neoadjuvant chemoradiation (CRT plus surgery). In the surgery group, pT1, 2, 3, and 4 disease was found in 15, 17, 104, and 34 patients, respectively; their respective pN1 rates were 13.3%, 29.4%, 57.7%, and 64.7%, P < 0.01. In CRT plus surgery, pT0, T1, 2, 3, and 4 were found in 48, 12, 23, 21, and 5 patients, respectively; their respective pN1 rates were 31.3%, 16.7%, 21.7%, 52.4%, and 20%, P = 0.44. Logistic regression analysis of factors predictive of pN1 showed that pT stage correlated with pN1 status (P = 0.005) in the surgery group, but not for the CRT plus surgery group. Cox regression analysis demonstrated that in the surgery group, pT, pN, and R category, and overall pTNM stage, were independent prognostic factors, whereas pN, R category, and gender were identified as relevant for CRT plus surgery. After chemoradiation, pT and overall pTNM stage groupings were not as clearly prognostic as in patients without prior therapy. Nodal status remains an important prognostic factor. PMID- 17114017 TI - Prospective trial of laparoscopic nissen fundoplication versus proton pump inhibitor therapy for gastroesophageal reflux disease: Seven-year follow-up. AB - Laparoscopic Nissen fundoplication and proton pump inhibitor (PPI) therapy are both established treatments for gastroesophageal reflux disease (GERD). We have performed a prospective randomized study comparing these two treatments and now have long-term follow-up data. Between July 1997 and August 2001, 183 patients in Norwich took part in a randomized controlled trial comparing laparoscopic Nissen fundoplication and PPI therapy for the treatment of GERD. In October 2005, patients were followed up and asked to complete a reflux symptom questionnaire. Ninety-one patients were randomized to have surgery and 92 to have optimized PPI therapy. After 12 months, those who had been randomized to PPI were offered the opportunity to have surgery. Fifty-four patients went on to have antireflux surgery; the remaining 38 did not. In all three groups, there was a significant improvement in symptom score after the initial 12 months (P < 0.01; Mann-Whitney U test). However, those who later had surgery despite having had optimal PPI treatment beforehand experienced further symptomatic improvement (P < 0.01) at long-term follow-up (median 6.9 years, range, 4.3-8.3). Both optimal PPI therapy and laparoscopic Nissen fundoplication are effective treatments for GERD. However, surgery offers additional benefit for those who have only partial symptomatic relief whilst on PPIs. PMID- 17114038 TI - Developing dendrites demonstrate unexpected specificity. AB - Our knowledge of how developing dendrites attain their mature state is still rudimentary. In this issue of Neuron, Mumm et al. rely on time-lapsed analysis of ingrowing dendrites of retinal ganglion cells in transgenic zebrafish to show that this process is much more specific than has been suspected. PMID- 17114039 TI - Dissecting the coupling between the voltage sensor and pore domains. AB - The gating mechanism of K(v) channels is not known. In this issue of Neuron, Soler-Llavina et al. present fascinating results that support the concept of relatively independent voltage-sensing modules. However, they also find that its interactions with the pore domain are rather complex, with specific S4-S5 intersubunit contacts underlying the concerted transition leading to the channel opening. PMID- 17114040 TI - Synaptic homeostasis on the fast track. AB - Synaptic homeostasis is a phenomenon that prevents the nervous system from descending into chaos. In this issue of Neuron, Frank et al. overturn the notion that synaptic homeostasis at Drosophila NMJs is a slow developmental process. They report that postsynaptic changes are offset within minutes by a homeostatic increase in neurotransmitter release that requires the presynaptic Ca(2+) channel Cacophony. PMID- 17114041 TI - A startling role for synaptic zinc. AB - It has long been known that the synaptic vesicles of certain glutamatergic terminals, as well as some inhibitory terminals, are richly supplied with zinc ions, yet the functional role of this pool of zinc in synaptic transmission has remained elusive. In this issue of Neuron, Hirzel et al. provide direct in vivo evidence that endogenous zinc is required for proper functioning of neuronal circuitry in the brainstem and spinal cord. They show that knockin mice carrying a point mutation which eliminates zinc potentiation of alpha1-containing glycine receptors develop severe sensorimotor deficits characteristic of impaired glycinergic neurotransmission. PMID- 17114042 TI - Maps, codes, and sequence elements: can we predict the protein output from an alternatively spliced locus? AB - Alternative splicing choices are governed by splicing regulatory protein interactions with splicing silencer and enhancer elements present in the pre mRNA. However, the prediction of these choices from genomic sequence is difficult, in part because the regulators can act as either enhancers or silencers. A recent study describes how for a particular neuronal splicing regulatory protein, Nova, the location of its binding sites is highly predictive of the protein's effect on an exon's splicing. PMID- 17114043 TI - Experience-dependent plasticity in adult visual cortex. AB - Experience-dependent plasticity is a prominent feature of the mammalian visual cortex. Although such neural changes are most evident during development, adult cortical circuits can be modified by a variety of manipulations, such as perceptual learning and visual deprivation. Elucidating the underlying mechanisms at the cellular and synaptic levels is an essential step in understanding neural plasticity in the mature animal. Although developmental and adult plasticity share many common features, notable differences may be attributed to developmental cortical changes at multiple levels. These range from shifts in the molecular profiles of cortical neurons to changes in the spatiotemporal dynamics of network activity. In this review, we will discuss recent progress and remaining challenges in understanding adult visual plasticity, focusing on the primary visual cortex. PMID- 17114044 TI - The familial Parkinsonism gene LRRK2 regulates neurite process morphology. AB - Mutations in LRRK2 underlie an autosomal-dominant, inherited form of Parkinson's disease (PD) that mimics the clinical features of the common "sporadic" form of PD. The LRRK2 protein includes putative GTPase, protein kinase, WD40 repeat, and leucine-rich repeat (LRR) domains of unknown function. Here we show that PD associated LRRK2 mutations display disinhibited kinase activity and induce a progressive reduction in neurite length and branching both in primary neuronal cultures and in the intact rodent CNS. In contrast, LRRK2 deficiency leads to increased neurite length and branching. Neurons that express PD-associated LRRK2 mutations additionally harbor prominent phospho-tau-positive inclusions with lysosomal characteristics and ultimately undergo apoptosis. PMID- 17114045 TI - Son of sevenless directly links the Robo receptor to rac activation to control axon repulsion at the midline. AB - Son of sevenless (Sos) is a dual specificity guanine nucleotide exchange factor (GEF) that regulates both Ras and Rho family GTPases and thus is uniquely poised to integrate signals that affect both gene expression and cytoskeletal reorganization. Here, using genetics, biochemistry, and cell biology, we demonstrate that Sos is recruited to the plasma membrane, where it forms a ternary complex with the Roundabout receptor and the SH3-SH2 adaptor protein Dreadlocks (Dock) to regulate Rac-dependent cytoskeletal rearrangement in response to the Slit ligand. Intriguingly, the Ras and Rac-GEF activities of Sos can be uncoupled during Robo-mediated axon repulsion; Sos axon guidance function depends on its Rac-GEF activity, but not its Ras-GEF activity. These results provide in vivo evidence that the Ras and RhoGEF domains of Sos are separable signaling modules and support a model in which Robo recruits Sos to the membrane via Dock to activate Rac during midline repulsion. PMID- 17114046 TI - In vivo imaging reveals dendritic targeting of laminated afferents by zebrafish retinal ganglion cells. AB - Targeting of axons and dendrites to particular synaptic laminae is an important mechanism by which precise patterns of neuronal connectivity are established. Although axons target specific laminae during development, dendritic lamination has been thought to occur largely by pruning of inappropriately placed arbors. We discovered by in vivo time-lapse imaging that retinal ganglion cell (RGC) dendrites in zebrafish show growth patterns implicating dendritic targeting as a mechanism for contacting appropriate synaptic partners. Populations of RGCs labeled in transgenic animals establish distinct dendritic strata sequentially, predominantly from the inner to outer retina. Imaging individual cells over successive days confirmed that multistratified RGCs generate strata sequentially, each arbor elaborating within a specific lamina. Simultaneous imaging of RGCs and subpopulations of presynaptic amacrine interneurons revealed that RGC dendrites appear to target amacrine plexuses that had already laminated. Dendritic targeting of prepatterned afferents may thus be a novel mechanism for establishing proper synaptic connectivity. PMID- 17114047 TI - Functional interactions at the interface between voltage-sensing and pore domains in the Shaker K(v) channel. AB - Voltage-activated potassium (K(v)) channels contain a central pore domain that is partially surrounded by four voltage-sensing domains. Recent X-ray structures suggest that the two domains lack extensive protein-protein contacts within presumed transmembrane regions, but whether this is the case for functional channels embedded in lipid membranes remains to be tested. We investigated domain interactions in the Shaker K(v) channel by systematically mutating the pore domain and assessing tolerance by examining channel maturation, S4 gating charge movement, and channel opening. When mapped onto the X-ray structure of the K(v)1.2 channel the large number of permissive mutations support the notion of relatively independent domains, consistent with crystallographic studies. Inspection of the maps also identifies portions of the interface where residues are sensitive to mutation, an external cluster where mutations hinder voltage sensor activation, and an internal cluster where domain interactions between S4 and S5 helices from adjacent subunits appear crucial for the concerted opening transition. PMID- 17114048 TI - Anomalous diffusion in Purkinje cell dendrites caused by spines. AB - We combined local photolysis of caged compounds with fluorescence imaging to visualize molecular diffusion within dendrites of cerebellar Purkinje cells. Diffusion of a volume marker, fluorescein dextran, within spiny dendrites was remarkably slow in comparison to its diffusion in smooth dendrites. Computer simulations indicate that this retardation is due to a transient trapping of molecules within dendritic spines, yielding anomalous diffusion. We considered the influence of spine trapping on the diffusion of calcium ions (Ca(2+)) and inositol-1,4,5-triphospate (IP(3)), two synaptic second messengers. Diffusion of IP(3) was strongly influenced by the presence of dendritic spines, while Ca(2+) was removed so rapidly that it could not diffuse far enough to be trapped. We conclude that an important function of dendritic spines may be to trap chemical signals and thereby create slowed anomalous diffusion within dendrites. PMID- 17114049 TI - State-dependent mechanisms of LTP expression revealed by optical quantal analysis. AB - The expression mechanism of long-term potentiation (LTP) remains controversial. Here we combine electrophysiology and Ca(2+) imaging to examine the role of silent synapses in LTP expression. Induction of LTP fails to change p(r) at these synapses but instead mediates an unmasking process that is sensitive to the inhibition of postsynaptic membrane fusion. Once unmasked, however, further potentiation of formerly silent synapses leads to an increase in p(r). The state of the synapse thus determines how LTP is expressed. PMID- 17114050 TI - Mechanisms underlying the rapid induction and sustained expression of synaptic homeostasis. AB - Homeostatic signaling systems are thought to interface with the mechanisms of neural plasticity to achieve stable yet flexible neural circuitry. However, the time course, molecular design, and implementation of homeostatic signaling remain poorly defined. Here we demonstrate that a homeostatic increase in presynaptic neurotransmitter release can be induced within minutes following postsynaptic glutamate receptor blockade. The rapid induction of synaptic homeostasis is independent of new protein synthesis and does not require evoked neurotransmission, indicating that a change in the efficacy of spontaneous quantal release events is sufficient to trigger the induction of synaptic homeostasis. Finally, both the rapid induction and the sustained expression of synaptic homeostasis are blocked by mutations that disrupt the pore-forming subunit of the presynaptic Ca(V)2.1 calcium channel encoded by cacophony. These data confirm the presynaptic expression of synaptic homeostasis and implicate presynaptic Ca(V)2.1 in a homeostatic retrograde signaling system. PMID- 17114051 TI - Hyperekplexia phenotype of glycine receptor alpha1 subunit mutant mice identifies Zn(2+) as an essential endogenous modulator of glycinergic neurotransmission. AB - Zn(2+) is thought to modulate neurotransmission by affecting currents mediated by ligand-gated ion channels and transmitter reuptake by Na(+)-dependent transporter systems. Here, we examined the in vivo relevance of Zn(2+) neuromodulation by producing knockin mice carrying the mutation D80A in the glycine receptor (GlyR) alpha1 subunit gene (Glra1). This substitution selectively eliminates the potentiating effect of Zn(2+) on GlyR currents. Mice homozygous for Glra1(D80A) develop a severe neuromotor phenotype postnatally that resembles forms of human hyperekplexia (startle disease) caused by mutations in GlyR genes. In spinal neurons and brainstem slices from Glra1(D80A) mice, GlyR expression, synaptic localization, and basal glycinergic transmission were normal; however, potentiation of spontaneous glycinergic currents by Zn(2+) was significantly impaired. Thus, the hyperekplexia phenotype of Glra1(D80A) mice is due to the loss of Zn(2+) potentiation of alpha1 subunit containing GlyRs, indicating that synaptic Zn(2+) is essential for proper in vivo functioning of glycinergic neurotransmission. PMID- 17114052 TI - Phospholipase Cbeta 3 mediates the scratching response activated by the histamine H1 receptor on C-fiber nociceptive neurons. AB - Phospholipase Cbeta (PLCbeta) isozymes represent a family of molecules that link G protein-coupled receptors (GPCRs) to an intracellular signaling network. Here, we investigated the function of PLCbeta isozymes in sensory neurons by using mutant mice deficient for specific PLCbeta family members. Expression analysis indicated that PLCbeta3, one of the four isoforms, is predominantly expressed in a subpopulation of C-fiber nociceptors. A subset of these neurons expressed the histamine H1 receptor. Ca(2+) imaging studies revealed that PLCbeta3 specifically mediates histamine-induced calcium responses through the histamine H1 receptor in cultured sensory neurons. In line with this, we found that PLCbeta3(-/-) mice showed significant defects in scratching behavior induced by histamine; histamine trifluoromethyl-toluidine (HTMT), a selective H1 agonist; and compound 48/80, a mast cell activator. These results demonstrate that PLCbeta3 is required to mediate "itch" sensation in response to histamine acting on the histamine H1 receptor in C-fiber nociceptive neurons. PMID- 17114053 TI - Nonmonotonic synaptic excitation and imbalanced inhibition underlying cortical intensity tuning. AB - Intensity-tuned neurons, characterized by their nonmonotonic response-level function, may play important roles in the encoding of sound intensity-related information. The synaptic mechanisms underlying intensity tuning remain unclear. Here, in vivo whole-cell recordings in rat auditory cortex revealed that intensity-tuned neurons, mostly clustered in a posterior zone, receive imbalanced tone-evoked excitatory and inhibitory synaptic inputs. Excitatory inputs exhibit nonmonotonic intensity tuning, whereas with tone intensity increments, the temporally delayed inhibitory inputs increase monotonically in strength. In addition, this delay reduces with the increase of intensity, resulting in an enhanced suppression of excitation at high intensities and a significant sharpening of intensity tuning. In contrast, non-intensity-tuned neurons exhibit covaried excitatory and inhibitory inputs, and the relative time interval between them is stable with intensity increments, resulting in monotonic response-level function. Thus, cortical intensity tuning is primarily determined by excitatory inputs and shaped by cortical inhibition through a dynamic control of excitatory and inhibitory timing. PMID- 17114054 TI - Backward shift of head direction tuning curves of the anterior thalamus: comparison with CA1 place fields. AB - The head direction cell system is composed of multiple regions associated with the hippocampal formation. The dynamics of head direction tuning curves (HDTCs) were compared with those of hippocampal place fields. In both familiar and cue altered environments, as a rat ran an increasing number of laps on a track, the center of mass (COM) of the HDTC tended to shift backward, similar to shifting observed in place cells. However, important differences existed between these cells in terms of the shift patterns relative to the cue-altered conditions, the proportion of backward versus forward shifts, and the time course of shift resetting. The demonstration of backward COM shifts in head direction cells and place cells suggests that similar plasticity mechanisms (such as temporally asymmetric LTP induction or spike timing-dependent plasticity) may be at work in both brain systems, and these processes may reflect a general mechanism for storing learned sequences of neural activity patterns. PMID- 17114055 TI - Modular organization of finger movements by the human central nervous system. AB - The motor system may generate automated movements, such as walking, by combining modular spinal motor synergies. However, it remains unknown whether a modular neuronal architecture is sufficient to generate the unique flexibility of human finger movements, which rely on cortical structures. Here we show that finger movements evoked by transcranial magnetic stimulation (TMS) of the primary motor cortex reproduced distinctive features of the spatial representation of voluntary movements as identified in previous neuroimaging studies, consistent with naturalistic activation of neuronal elements. Principal component analysis revealed that the dimensionality of TMS-evoked movements was low. Principal components extracted from TMS-induced finger movements resembled those derived from end-postures of voluntary movements performed to grasp imagined objects, and a small subset of them was sufficient to reconstruct these movements with remarkable fidelity. The motor system may coordinate even the most dexterous movements by using a modular architecture involving cortical components. PMID- 17114056 TI - Mapping the phosphoinositide-binding site on chick cofilin explains how PIP2 regulates the cofilin-actin interaction. AB - Cofilin plays a key role in the choreography of actin dynamics via its ability to sever actin filaments and increase the rate of monomer dissociation from pointed ends. The exact manner by which phosphoinositides bind to cofilin and inhibit its interaction with actin has proven difficult to ascertain. We determined the structure of chick cofilin and used NMR chemical shift mapping and structure directed mutagenesis to unambiguously locate its recognition site for phosphoinositides (PIs). This structurally unique recognition site requires both the acyl chain and head group of the PI for a productive interaction, and it is not inhibited by phosphorylation of cofilin. We propose that the interaction of cofilin with membrane-bound PIs abrogates its binding to both actin and actin interacting protein 1, and facilitates spatiotemporal regulation of cofilin activity. PMID- 17114057 TI - Evaluation of a diffusion-driven mechanism for substrate ubiquitination by the SCF-Cdc34 ubiquitin ligase complex. AB - Release of ubiquitin-charged Cdc34 from the SCF ubiquitin ligase followed by diffusion-driven collision with substrate has been proposed to underlie ubiquitination of the canonical SCF substrate Sic1. Cdc34 F72V, reported to be defective in dissociation from SCF, served as key validation. Here, we test predictions of this "hit-and-run" hypothesis. We find that Cdc34 F72V is generally defective in SCF-mediated activation but, contrary to expectation, does not compete with wild-type Cdc34 in vitro or in vivo and can fulfill the physiological role of Cdc34 with only moderate delay in Sic1 turnover. Whereas a hit-and-run mechanism might explain how Cdc34 can transfer ubiquitin to the ends of growing ubiquitin chains on SCF-bound substrates, molecular modeling suggests that an E2 docked to SCF can do so without dissociating. We propose that interactions between Cdc34 approximately Ub and SCF directly activate ubiquitin transfer within a substrate-SCF-Cdc34 approximately Ub ternary complex. PMID- 17114058 TI - RNA polymerase pausing regulates translation initiation by providing additional time for TRAP-RNA interaction. AB - RNA polymerase (RNAP) pause sites have been identified in several prokaryotic genes. Although the presumed biological function of RNAP pausing is to allow synchronization of RNAP position with regulatory factor binding and/or RNA folding, a direct causal link between pausing and changes in gene expression has been difficult to establish. RNAP pauses at two sites in the Bacillus subtilis trpEDCFBA operon leader. Pausing at U107 and U144 participates in transcription attenuation and trpE translation control mechanisms, respectively. Substitution of U144 caused a substantial pausing defect in vitro and in vivo. These mutations led to increased trp operon expression that was suppressed by overproduction of TRAP, indicating that pausing at U144 provides additional time for TRAP to bind to the nascent transcript and promote formation of an RNA structure that blocks translation of trpE. These results establish that pausing is capable of playing a role in regulating translation in bacteria. PMID- 17114059 TI - Architecture of the 99 bp DNA-six-protein regulatory complex of the lambda att site. AB - The highly directional and tightly regulated recombination reaction used to site specifically excise the bacteriophage lambda chromosome out of its E. coli host chromosome requires the binding of six sequence-specific proteins to a 99 bp segment of the phage att site. To gain structural insights into this recombination pathway, we measured 27 FRET distances between eight points on the 99 bp regulatory DNA bound with all six proteins. Triangulation of these distances using a metric matrix distance-geometry algorithm provided coordinates for these eight points. The resulting path for the protein-bound regulatory DNA, which fits well with the genetics, biochemistry, and X-ray crystal structures describing the individual proteins and their interactions with DNA, provides a new structural perspective into the molecular mechanism and regulation of the recombination reaction and illustrates a design by which different families of higher-order complexes can be assembled from different numbers and combinations of the same few proteins. PMID- 17114060 TI - SeqA blocking of DnaA-oriC interactions ensures staged assembly of the E. coli pre-RC. AB - DnaA occupies only the three highest-affinity binding sites in E. coli oriC throughout most of the cell cycle. Immediately prior to initiation of chromosome replication, DnaA interacts with additional recognition sites, resulting in localized DNA-strand separation. These two DnaA-oriC complexes formed during the cell cycle are functionally and temporally analogous to yeast ORC and pre-RC. After initiation, SeqA binds to hemimethylated oriC, sequestering oriC while levels of active DnaA are reduced, preventing reinitiation. In this paper, we investigate how resetting of oriC to the ORC-like complex is coordinated with SeqA-mediated sequestration. We report that oriC resets to ORC during sequestration. This was possible because SeqA blocked DnaA binding to hemimethylated oriC only at low-affinity recognition sites associated with GATC but did not interfere with occupation of higher-affinity sites. Thus, during the sequestration period, SeqA repressed pre-RC assembly while ensuring resetting of E. coli ORC. PMID- 17114061 TI - PET tracer development--a tale of mice and men. AB - PET scanning is an emerging technology for the clinical evaluation of many disease processes in man. The vast majority of clinical positron emission tomography (PET) studies are performed using a single tracer, fluorodeoxyglucose. Despite the excellent diagnostic performance of this tracer, it has recognised limitations. New tracers offer the potential to both address these limitations, and to establish new applications for PET. Small animal PET is a logical technique for validating new tracers relevant to human diseases. However, interspecies differences in the handling of chemicals may significantly influence the handling of novel tracers. This requires caution in extrapolating findings in animals to expectations of performance in man. Already there are several examples where biodistribution studies in mice would not have predicted the clinical utility of existing PET tracers. Nevertheless, application of a systematic approach to tracer development is likely to speed transition of new tracers from animals into man. PMID- 17114062 TI - From anatomical to biological target volumes: the role of PET in radiation treatment planning. AB - Progress in radiation oncology requires a re-evaluation of the methods of target volume delineation beyond anatomical localization. New molecular imaging techniques for tumour visualisation such as positron emission tomography (PET) provide insight into tumour characteristics and can be complementary to the anatomical data of computed tomography or magnetic resonance imaging. In this review, three issues are discussed: First, can PET identify a tumour more accurately? Second, can biological tumour characteristics be visualised? Third, can intratumoural heterogeneity of these characteristics be identified? PMID- 17114063 TI - PET imaging of tumour hypoxia. AB - Tumour hypoxia represents a significant challenge to the curability of human tumours leading to treatment resistance and enhanced tumour progression. Tumour hypoxia can be detected by non-invasive and invasive techniques but the inter relationships between these remains largely undefined. [18F]Fluoromisonidazole-3 fluoro-1-(2'-nitro-1'-imidazolyl)-2-propanol ([18F]MISO) and Cu-diacetyl-bis(N4 methylthiosemicarbazone (Cu-ATSM)-positron emission tomography (PET), and blood oxygen level-dependent (BOLD)-magnetic resonance imaging (MRI) are the lead contenders for human application based on their non-invasive nature, ease of use and robustness, measurement of hypoxia status, validity, ability to demonstrate heterogeneity and general availability; PET techniques are the primary focus of this review. PMID- 17114065 TI - Imaging response assessment in oncology. AB - The role of imaging in the clinical setting as well as in the drug development process is expanding rapidly. Imaging technology now exists that is capable of detecting tumor response within hours. In parallel with this advance, a new array of more targeted and specific therapies are being developed. This paradigm shift in turn demands a more sophisticated way of quantifying response. There is a need to update and modify the current response evaluation criteria in solid tumors (RECIST), which rely solely on anatomic size measurement of tumors. In addition, response assessment guidelines will need to be increasingly disease-specific. Response assessment by imaging is now intimately involved with all stages of the drug development process, from exploratory drug discovery through clinical trials, as well as in clinical use. Imaging biomarkers and surrogate endpoints have the potential to speed drug approval significantly. The major funding institutions and the pharmaceutical industry are working more and more with researchers to help maintain progress in this multidisciplinary area involving oncologists, radiologists, molecular imaging specialists, medical physicists, and computer scientists. PMID- 17114066 TI - Screening for colon cancer. AB - Colorectal cancer screening reduces mortality in individuals 50 years and older. Each of the screening tests currently available has advantages and limitations, and there is no consensus as to which test or combination of tests is best. What is clear, however, is that the rates of colorectal cancer screening remain low. This review summarizes the clinical evidence supporting colorectal cancer screening in the average risk population and in high risk groups, discusses the advantages and disadvantages of the available screening tests, outlines the currently recommended guidelines for screening based on risk category, and discusses new and emerging technologies for colorectal cancer screening. PMID- 17114067 TI - Radiation injury: imaging findings in the chest, abdomen and pelvis after therapeutic radiation. AB - Radiation may be used as adjuvant or primary therapy in a variety of tumors in the chest, abdomen and pelvis. Therapeutic radiation affects not only malignant tumors but also surrounding normal tissues. The risk of injury depends on the size, number and frequency of radiation fractions, volume of irradiated tissue, duration of treatment, and method of radiation delivery. Concomitant chemotherapy can act synergistically to produce injury. Other predisposing factors include infection, prior surgery and chronic illness like hypertension, diabetes mellitus and atherosclerosis. Radiation changes vary, based on the target organ and the time from completion of therapy. While most serious complications related to radiotherapy are relatively uncommon, given the number of patients that are treated and the relatively long latency period for development of radiation changes, follow-up imaging studies frequently have findings that should be recognized as radiation related. Familiarity with the spectrum of imaging findings after radiation injury permits differentiation from other etiologies such as recurrent malignancy. The following will discuss imaging findings that may be seen during imaging surveillance in patients with malignancy affecting the chest, abdomen and pelvis. PMID- 17114068 TI - Imaging tumour motion for radiotherapy planning using MRI. AB - Novel technology has made dynamic magnetic resonance imaging (MRI) of lung motion and lung tumour mobility during continuous respiration feasible. This might be beneficial for planning of radiotherapy of lung tumours, especially when using high precision techniques. This paper describes the recent developments to analyze and visualize pulmonary nodules during continuous respiration using MRI. Besides recent dynamic two-dimensional approaches to quantify motion of pulmonary nodules during respiration novel three-dimensional techniques are presented. Beyond good correlation to pulmonary function tests MRI also provides regional information about differences between tumour-bearing and non-tumour bearing lung and the restrictive effects of radiotherapy as well as the compensation by the contralateral lung. PMID- 17114069 TI - Head and neck cancer: how imaging predicts treatment outcome. AB - Sophisticated imaging methods, such as computed tomography, magnetic resonance imaging and positron emission tomography, play an increasingly important role in the management of head and neck cancer. Pretreatment imaging findings have predictive value for patient outcome, independently from the currently used TNM classification, and may be used to tailor treatment to the individual patient. Based on per-treatment imaging, individualised replanning during radiotherapy may ameliorate tumour control rates and reduce toxic effects to normal tissues. Early posttreatment imaging studies contain important prognostic information, and allow selection of patients for further treatment or watchful waiting. PMID- 17114072 TI - CT/MRI of neuroendocrine tumours. AB - Neuroendocrine tumours (NETs) are often thought to be rare and rather recherche cancers which are of little concern to the general physician, surgeon or radiologist because of their rarity and esoteric nature. In fact, while relatively uncommon, the total group of gastro-entero-pancreatic (GEP) tumours incorporates the spectrum of all types of carcinoids, including bronchial carcinoids, and the whole gamut of islet-cell tumours. Some of these may present as functioning tumours, with a plethora of hormonal secretions and concomitant clinical syndromes, and GEPs in general have an incidence around 30 per million population per year. This means that in the whole European Union, for example, there will be in the region of 12,000 new patients every year presenting with one or another manifestation of these tumours. Furthermore, the comparatively long survival of many of these patients, compared to more common adenocarcinomas or epithelial tumours, implies that the point prevalence is also not inconsiderable. However, it is undoubtedly true that these tumours can be difficult to identify, especially in their early stages, and it is then that radiological investigation becomes of paramount importance. Having taken into account all these considerations, most investigators would initiate investigation of a suspected or biochemically proven islet-cell tumour with cross-sectional imaging-either CT or MRI. This will clearly identify the larger lesions, allow assessment of the entire abdomen, and provide valuable information on the presence of hepatic metastates. PMID- 17114075 TI - Imaging of the spine in patients with malignancy. AB - This contribution presents an approach to the diagnosis of symptoms referable to spinal pathology in patients with known malignancy. Pain and neurological disturbance are distressing and disabling symptoms, which in patients with cancer may be a result of bony metastases, paraspinal soft tissue disease and meningeal and intra-axial spinal metastases. Imaging studies are pivotal, and typical and atypical imaging features are presented. PMID- 17114073 TI - Nuclear medicine imaging and therapy of neuroendocrine tumours. AB - Radiolabelled peptides are used for specific targeting of receptors (over )expressed by tumour cells. Dependent on the kind of labelling and the radionuclide used, these compounds may be utilised for imaging or for therapy. A concise overview is provided on basic principles of designing and developing radiopeptides for these applications. Furthermore, clinical application of these compounds for imaging and therapy is described. Advantages of the method compared to other techniques (such as the use of radiolabelled antibodies or antibody fragments) are discussed as well as pitfalls and limitations. PMID- 17114076 TI - The orbits in cancer imaging. AB - Primary malignant lesions in the orbit are relatively uncommon. However, the orbits are frequently involved in haematogeneous metastasis or by direct extension from malignancies originating from the adjacent nasal cavity or paranasal sinuses. This paper focuses on the more commonly encountered primary orbital malignancies and the mapping of tumour spread into the orbits. PMID- 17114077 TI - Solitary pulmonary nodule: detection and management. AB - Pulmonary nodules are commonly detected at computed tomography (CT) of the chest. More than 95% are < or = 10 mm; of these more than 95% are benign. Visual detection of pulmonary nodules by human readers is suboptimal, particularly with small nodules < or =10 mm. Computer-assisted detection can improve sensitivity and diagnostic confidence. Due to the high proportion of malignant lesions in nodules >10 mm immediate, often invasive workup is required including contrast enhanced dynamic CT, positron emission tomography (PET) or biopsy. However, in nodules < or =10 mm the high proportion of benign lesions requires a non-invasive work-up usually based on follow-up with unenhanced CT. Invasive procedures are only required for growing nodules. Stable nodules require further follow-up and decreasing nodules are considered benign. PMID- 17114079 TI - PET/CT: will it change the way that we use CT in cancer imaging? AB - Accurate staging of cancer is of fundamental importance to treatment selection and planning. Current staging paradigms focus, first, on a detailed delineation of the primary tumour in order to determine its suitability for resection, and, thereafter, on assessment of the presence of metastatic spread that would alter the surgical approach, or mandate non-surgical therapies. This approach has, at its core, the assumption that the best, and sometimes the only, way to cure a patient of cancer is by surgical resection. Unfortunately, all non-invasive techniques in current use have imperfect ability to identify those primary tumours that are able to be completely excised, and even worse ability to define the extent of metastatic spread. Nevertheless, because of relatively low cost and widespread availability, computed tomography (CT) scanning is the preferred methodology for tumour, nodal and systemic metastasis (TNM) staging. This is often supplemented by other tests that have improved performance in particular staging domains. For example, magnetic resonance imaging (MRI), mammography, or endoscopic ultrasound may be used as complementary tests for T-staging; surgical nodal sampling for N-staging; and bone scanning, MRI or ultrasound for M-staging. Accordingly, many patients undergo a battery of investigations but, even then, are found to have been incorrectly staged based on subsequent outcomes. Even for those staged surgically, pathology can only identify metastases within the resection specimens and has no capability for detecting remote disease. As a result of this, many patients undergo futile operations for disease that could never have been cured by surgery. In the case of restaging, the situation is even worse. The sequelae of prior treatment can be difficult to differentiate from residual cancer and the likelihood of successful salvage therapy is even less than at presentation. More deleteriously, patients may be subjected to additional morbid treatments when cure has already been achieved. Thus, in post-treatment follow-up, the presence and extent of disease is equally critical to treatment selection and patient outcome as it is in primary staging. One of the major strengths of positron emission tomography (PET)/CT as a cancer staging modality is its ability to identify systemic metastases. At any phase of cancer evaluation, demonstration of systemic metastasis has profound therapeutic and prognostic implications. Only in the absence of systemic metastasis does nodal status become important, and only when unresectable nodal metastasis has been excluded does T-stage become important. There are now accumulating data that PET/CT could be used as the first, rather than the last test to assess M- and N stage for evaluating cancers with an intermediate to high pre-test likelihood of metastatic disease based on poor long-term survival. In this scenario, there is great opportunity for subsequently selecting and tailoring the performance of anatomically based imaging modalities to define the structural relations of abnormalities identified by PET, when this information would be of relevance to management planning. Primary staging of oesophageal cancer and restaging of colorectal cancer are illustrative examples of a new paradigm for cancer imaging. PMID- 17114080 TI - PET in lymphoma. AB - This review attempts to discuss the role of positron emission tomography (PET) imaging for staging, treatment response and follow-up of patients with lymphoma. The pitfalls and impact of PET imaging on the clinical management are also addressed. PMID- 17114081 TI - FDG-PET in colorectal cancer. AB - [18F]Fluorodeoxyglucose (FDG) positron emission tomography (PET) is a useful imaging tool in the evolving management of patients with colorectal carcinoma. This technique is able to measure and visualize metabolic changes in cancer cells. This feature results in the ability to distinguish viable tumor from scar tissue, in the detection of tumor foci at an earlier stage than possible by conventional anatomic imaging and in the measurement of alterations in tumor metabolism, indicative of tumor response to therapy. Nowadays, FDG-PET plays a pivotal role in staging patients before surgical resection of recurrence and metastases, in the localization of recurrence in patients with an unexplained rise in serum carcinoembryonic antigen and in assessment of residual masses after treatment. In the presurgical evaluation, FDG-PET may be best used in conjunction with anatomic imaging in order to combine the benefits of both anatomical (CT) and functional (PET) information, which leads to significant improvements in preoperative liver staging and preoperative judgment on the feasibility of resection. Integration of FDG-PET into the management algorithm of these categories of patients alters and improves therapeutic management, reduces morbidity due to futile surgery, leads to substantial cost savings and probably also to a better patient outcome. FDG-PET also appears to have great potential in monitoring the success of local ablative therapies soon after intervention and in the prediction and evaluation of response to radiotherapy, systemic therapy, and combinations thereof. This review aims to outline the current and future role of FDG-PET in the field of colorectal cancer. PMID- 17114082 TI - PET: other thoracic malignancies. AB - The vast majority of esophageal cancers are fluorodeoxyglucose (FDG) avid; the primary use for positron emission tomography (PET) in patients with esophageal cancer is in the detection of distant metastases, because known distant metastatic disease precludes surgical resection. High standardized uptake values (SUVs) may be predictive of poor prognosis. PET findings may be used to assess therapy response and evaluate for esophageal tumor recurrence after treatment. PET findings may be non-specific in different types of thymic lesions, although thymic carcinomas tend to be extremely FDG avid. PET can be helpful in detecting distant spread from invasive thymomas and thymic carcinomas. Similarly, PET may be used to assess the extent of disease in patients with malignant pleural mesothelioma, thereby facilitating optimal therapy approaches. PMID- 17114083 TI - PET in face and neck tumours. AB - FDG-PET is a useful tool in the imaging of head and neck tumours. It can be used to stage the primary tumour, to assess response to therapy and most importantly for the detection of recurrent tumour. The advantages and limitations of this technique are discussed in this article. PMID- 17114084 TI - Screening for lung cancer. AB - The lethality of lung cancer is related to the advanced stage at diagnosis. Initial studies have demonstrated that screening computed tomography (CT) is effective in diagnosing lung cancer at an earlier stage when compared with current clinical practice, however the best clinical approach for screening detected nodules has to be defined. The population to be identified as high risk should be over 50 years of age and should have smoked at least one pack/day for 20 years. CT protocols should use multidetector CT, low dose and a 2.5 reconstruction interval. Diagnostic workup on detected nodules should be designed according to size and consider CT at 3 or 12 months to evaluate doubling time, CT enhancement, PET/CT and/or FNAB or VATS. The prevalence of lung cancer in the screened population is 1.1%-2.7%, and the incidence is 0.2%-1.1%. Eighty-one percent of cancers are diagnosed in stage I. The percentage of surgery performed for benign lesions ranges from 21% to 55%. In our series, the overall mortality rate was 3.2% in 5 years. The results of randomized clinical studies, when available, will assess the real efficacy of CT in reducing lung cancer related mortality. PMID- 17114086 TI - Use and misuse of motor-vehicle crash death rates in assessing highway-safety performance. AB - OBJECTIVES: The objectives of the article are to assess the extent to which comparisons of motor-vehicle crash death rates can be used to determine the effectiveness of highway-safety policies over time in a country or to compare policy effectiveness across countries. METHODS: Motor-vehicle crash death rates per mile traveled in the 50 U.S. states from 1980 to 2003 are used to show the influence on these rates of factors independent of highway-safety interventions. Multiple regression models relating state death rates to various measures related to urbanization and demographics are used. RESULTS: The analyses demonstrate strong relationships between state death rates and urbanization and demographics. Almost 60% of the variability among the state death rates can be explained by the independent variables in the multiple regression models. When the death rates for passenger vehicle occupants (i.e., excluding motorcycle, pedestrian, and other deaths) are used in the regression models, almost 70% of the variability in the rates can be explained by urbanization and demographics. CONCLUSIONS: The analyses presented in the article demonstrate that motor-vehicle crash death rates are strongly influenced by factors unrelated to highway-safety countermeasures. Overall death rates should not be used as a basis for judging the effectiveness (or ineffectiveness) of specific highway-safety countermeasures or to assess overall highway-safety policies, especially across jurisdictions. There can be no substitute for the use of carefully designed scientific evaluations of highway-safety interventions that use outcome measures directly related to the intervention; e.g., motorcyclist deaths should be used to assess the effectiveness of motorcycle helmet laws. While this may seem obvious, there are numerous examples in the literature of death rates from all crashes being used to assess the effectiveness of interventions aimed at specific subsets of crashes. PMID- 17114087 TI - Effects of electronic stability control: an update. AB - OBJECTIVE: An earlier study reported that electronic stability control (ESC) in passenger vehicles reduced single-vehicle crash involvement risk by 41% and single-vehicle fatal crash involvement risk by 56%. The purpose of the present study was to update these effectiveness estimates using an additional year of crash data and a larger set of vehicle models. METHODS: The amount of data increased by half, allowing for separate effectiveness estimates for cars and sport utility vehicles (SUVs) and a more detailed examination of multiple-vehicle crash types. Crash involvement rates per registered vehicle were compared for otherwise identical vehicle models with and without ESC. RESULTS: Based on all police-reported crashes in 10 states during three years, ESC reduced single vehicle crash involvement risk by approximately 41%. Effects were significantly higher for SUVs than for cars. ESC reduced single-vehicle crash involvement risk by 49% for SUVs and 33% for cars. Based on all fatal crashes in the United States during four years, ESC was found to have reduced single-vehicle fatal crash involvement risk by 56%. Again, effectiveness estimates were higher for SUVs than for cars--59% for SUVs and 53% for cars, but these differences were not statistically significant. Multiple-vehicle fatal crash involvement risk was reduced by 32%-37% for SUVs and 25% for cars. CONCLUSIONS: The present study confirms the results of the earlier study. There are significant reductions in single-vehicle crash rates when passenger vehicles are equipped with ESC. In addition, ESC leads to reductions in severe multiple-vehicle crashes. PMID- 17114088 TI - Relationship between traffic fatalities and drunk driving in Japan. AB - OBJECTIVE: The present study was performed to clarify the relation between alcohol use and traffic fatalities in accidents involving motor vehicles in Japan. METHODS: Data on traffic accidents were collected from Fukuoka Prefectural Police records of traffic accidents which occurred in that prefecture between 1987 and 1996. Multiple logistic regression models were used to assess the effect of alcohol use on the risk of traffic-accident death. RESULTS: The data showed that 58,421 male drivers were involved in traffic accidents during the 10-year study period, and that 271 of these were killed as a result of the accident. Alcohol use was significantly associated with speed, seat belt use, time, and road form. Among male motorcar drivers, the odds ratio of alcohol use before driving, after adjusting for age, calendar year, time, and road form, was 4.08 (95% confidence interval, 3.08-5.40), which means that about 75% of fatalities (attributable risk percent among exposed) might have been prevented if drivers had not drunk before driving. CONCLUSIONS: Alcohol use before driving resulted in a 4.08-fold increase in the risk of death in a traffic accident. It is suggested that alcohol use is considered an important risk factor for fatality in traffic accidents. PMID- 17114089 TI - The role of personality characteristics in young adult driving. AB - BACKGROUND: Motor vehicle injury is the major cause of mortality among young adults. Information about the individual characteristics of those who drive dangerously could enhance traffic safety programs. The goal of this research was to examine the association between various personality-related characteristics and risky driving behaviors. METHODS: Young adults in Michigan, USA (n = 5,362) were surveyed by telephone regarding several personality factors (risk-taking, hostility, aggression, tolerance of deviance, achievement expectations) and driving behaviors (competitive driving, risk-taking driving, high-risk driving, aggressive driving, and drink/driving). Michigan driver records were obtained to examine offenses, serious offenses, driving offense points, crashes and serious crashes in the three pre-interview years. Multivariate regression analyses, adjusting for age, race, and marital status were conducted separately by sex to identify personality factors related to driving. RESULTS: For men and women, greater risk-taking propensity, physical/verbal hostility, aggression, and tolerance of deviance were significant predictors of a competitive attitude toward driving, risk-taking driving, high-risk driving, driving aggression, and drink/driving. Greater risk-taking propensity, physical/verbal hostility, aggression, and to a small degree, expectations for achievement predicted higher numbers of offenses, serious offenses, and points. CONCLUSION: Traffic safety policies and programs could be enhanced through recognition of the role personality factors play in driving behavior and the incorporation of this knowledge into the design and implementation of interventions that modify the behaviors associated with them. PMID- 17114090 TI - Trends over time in the risk of driver death: what if vehicle designs had not improved? AB - OBJECTIVES: Passenger vehicle driver death rates per million vehicle registrations declined steadily during calendar years 1985-2004. The present study sought to separate the effect of vehicle design changes from this trend. METHODS: Restricting the trend to a fixed set of model years removed the vehicle design effects, but there were still effects due to vehicle aging. Risk of driver death was found to increase each year vehicles aged, probably due to changes in vehicle use patterns. RESULTS: After separating out the vehicle design effects and making adjustments for the vehicle age effects, a different picture emerged of trends in death rates over time. Absent the vehicle design changes, the historical decline in driver fatality risk would have ended in 1993, with risk climbing ever since. This underlying trend has been obscured by changes in the vehicle fleet. CONCLUSIONS: The push for vehicle improvements has been worthwhile and can be credited with saving thousands of lives. However, the analysis shows that the gains in occupant protection from vehicle design improvements have been offset partially by an increasingly risky environment in recent years. Therefore, more attention needs to be paid to programs targeting improvement in roadway design and driver behavior. PMID- 17114091 TI - Determining older driver crash responsibility from police and insurance data. AB - This study aimed to determine the extent to which older drivers can be considered responsible for their crashes, to identify key factors in those crashes for which older drivers have been judged responsible, and to assess the extent to which older drivers' extra crash responsibility contributes to the road toll. Insurance claims from the State of Tasmania, Australia, for 1998-2002 were linked with police records for crashes involving drivers aged either 41-55 years or 65 years or older. Insurance and police data sets contained independent judgments of crash responsibility. There was a high level of agreement between the two sets of judgments, with older drivers judged around 1.5 times more likely to be responsible for their crashes than middle-aged drivers and, conversely, older drivers were around 0.6 as likely to be absolved from crash responsibility. It was concluded that older drivers' additional crash responsibility while valuable in explaining "what went wrong," currently makes only a small contribution to the overall road toll. PMID- 17114092 TI - A cost-benefit analysis of risk-reduction strategies targeted at older drivers. AB - OBJECTIVE: The risk of motor-vehicle collisions increases as driving-related functional abilities decline. These declines can accompany normal or pathological aging and can be identified through driving-related functional screening exams upon license renewal. The objective of this cost-benefit analysis was to determine the utility of four functional screening procedures used to identify drivers at risk for motor-vehicle collisions, as well as an intervention designed to maintain or improve functional abilities. Additionally, this study sought to determine the expected cost per driver if an intervention was designed to target only those drivers who failed the functional ability-based driving screen, versus the expected cost per driver if the intervention was distributed en masse to all drivers 75 years and older. Improving functional abilities in older adults has potential far-reaching health and financial impacts which are broader than their impact of maintaining mobility. METHODS: A decision tree was constructed to evaluate the expected costs and benefits of (a) screening all drivers and intervening when indicated (several screening batteries of varying length were considered), (b) no screening, but intervening with all drivers of older age, or (c) neither screening nor intervening (i.e., re-licensing per usual). Test characteristics and risk probabilities were based on a cohort of drivers aged 75 and older from a previous study (Ball et al., 2006). Relevant sensitivity analyses were conducted. RESULTS: Providing all drivers with the speed-of processing intervention is the most cost-beneficial option (expected cost per driver = $493.30), even if the cost of the intervention doubles. Sensitivity analysis indicated the effectiveness of the intervention could drop from 86% to 25% and the preventative approach of intervening with all drivers remains the most cost-beneficial strategy. The least cost-beneficial option is almost always re-licensing per usual (expected cost per driver = $1,562.84). CONCLUSION: Screening drivers upon license renewal is not currently beneficial because the available technology cannot consistently identify drivers at risk for a collision. However, the speed-of-processing intervention has demonstrated efficacy in improving driving competence (Roenker et al., 2003) and is a non invasive, moderate-cost intervention that has the potential to protect the safety and mobility, as well as the financial interests, of older drivers and the community at large. PMID- 17114093 TI - The impact of chronic pain patients' psychotropic drug knowledge and warning labels on the decision whether to drive a car or not. AB - OBJECTIVE: The attitudes of patients towards driving a car while taking medication with psychotropic side effects is unclear. A growing number of patients use these psychotropic medicines on a daily basis, and this may interfere with their ability to drive a car. METHODS: By means of a survey, we examined attitudes towards driving while using psychotropic medicinal drugs and the effect of warning labels on the decision whether to drive a car or not in patients with chronic pain. RESULTS: Fifty-eight of 100 patients possessing a driver's license used psychotropic medication. Despite warning labels affixed on the packages that these drugs might impair driving ability, the majority (71%) of these patients continued driving a car. A point of concern is that 40% of these patients reported not to be more cautious in traffic after taking psychotropic drugs. CONCLUSION: The results of this survey indicate that drug warning labels applied by Dutch pharmacies do not significantly change attitudes towards driving a car in patients taking medicinal drugs with psychotropic side effects. Future road-safety campaigns should pay more attention to the impairing effects of psychotropic drugs on driving. PMID- 17114094 TI - The effect of driver eye height on speed choice, lane-keeping, and car-following behavior: results of two driving simulator studies. AB - OBJECTIVE: Two simulator studies were conducted that assessed the effect of driver eye height on speed choice, lane-keeping, and car-following behavior. The effect of eye height on the subjective variables of mental workload, frustration, and confidence was also investigated, as was the contribution of drivers' aggression. METHODS: A total of 43 participants drove a simulated route while seated at two different eye heights: one that represented the view of the road from a large SUV and one that represented the view of the road from a small sports car. Driving scenarios were comprised of both open road and car-following segments. Dependent variables included driver-selected speed, speed variability, lane position, following distance to a slower-moving lead vehicle, and the subjective variables of frustration, confidence, and mental workload. RESULTS: When viewing the road from a high eye height, drivers drove faster, with more variability, and were less able to maintain a consistent position within the lane than when viewing the road from a low eye height. Driver eye height did not influence following distance to a slower-moving lead vehicle. Driver aggression had no effect on any of the dependent variables except level of frustration. CONCLUSIONS: The two studies demonstrate that, when they are not able to reference a speedometer, drivers choose to drive faster when they view the road from an eye height that is representative of a large SUV compared to that of a small sports car. There is a need to educate drivers of SUVs and other tall vehicles of this perceptual phenomenon in order to prevent collisions that may occur in conditions where it is impossible for drivers to base their speed selection solely on posted speed limits, such as in inclement weather. PMID- 17114095 TI - Parent driver characteristics associated with sub-optimal restraint of child passengers. AB - OBJECTIVES: To identify parent driver demographic and socioeconomic characteristics associated with the use of sub-optimal restraints for child passengers under nine years. METHODS: Cross-sectional study using in-depth, validated telephone interviews with parent drivers in a probability sample of 3,818 vehicle crashes involving 5,146 children. Sub-optimal restraint was defined as use of forward-facing child safety seats for infants under one or weighing under 20 lbs, and any seat-belt use for children under 9. RESULTS: Sub-optimal restraint was more common among children under one and between four and eight years than among children aged one to three years (18%, 65%, and 5%, respectively). For children under nine, independent risk factors for sub-optimal restraint were: non-Hispanic black parent drivers (with non-Hispanic white parents as reference, adjusted relative risk, adjusted RR = 1.24, 95% CI: 1.09 1.41); less educated parents (with college graduate or above as reference: high school, adjusted RR = 1.27, 95% CI: 1.12-1.44; less than high school graduate, adjusted RR = 1.36, 95% CI: 1.13-1.63); and lower family income (with $50,000 or more as reference: <$20,000, adjusted RR = 1.23, 95% CI: 1.07-1.40). Multivariate analysis revealed the following independent risk factors for sub-optimal restraint among four-to-eight-year-olds: older parent age, limited education, black race, and income below $20,000. CONCLUSIONS: Parents with low educational levels or of non-Hispanic black background may require additional anticipatory guidance regarding child passenger safety. The importance of poverty in predicting sub-optimal restraint underscores the importance of child restraint and booster seat disbursement and education programs, potentially through Medicaid. PMID- 17114096 TI - A new challenge to child and adolescent survival in urban Africa: an increasing burden of road traffic injuries. AB - OBJECTIVES: To develop an epidemiological profile and present the burden of urban road-traffic injuries (RTI) in children and adolescents in urban Sub-Saharan Africa. METHOD: A systematic review of published and gray literature of children and adolescents < or =19 years old involved in RTI in urban regions in Sub Saharan Africa from January 1980-December 2003. RESULTS: The mean annual incidence for urban road-traffic injuries was 109.8 per 100,000 children, and males were twice as involved as their female counterparts. Children between 10 to 14 years constitute the most frequent group involved in road crashes (36%) while pedestrians represent an average of 68% of all childhood RTI cases. Twenty Healthy Life Years per 1,000 children and adolescents are being lost annually in the region from RTI. CONCLUSIONS: There is an urgent need to develop a regional health research agenda to generate an accurate estimate of the burden of road traffic injuries on children. This is a new challenge for child health and there is a need to raise awareness among policy makers to promote appropriate interventions. PMID- 17114097 TI - Cervical spine loads and intervertebral motions during whiplash. AB - OBJECTIVE: To quantify the dynamic loads and intervertebral motions throughout the cervical spine during simulated rear impacts. METHODS: Using a biofidelic whole cervical spine model with muscle force replication and surrogate head and bench-top mini-sled, impacts were simulated at 3.5, 5, 6.5, and 8 g horizontal accelerations of the T1 vertebra. Inverse dynamics was used to calculate the dynamic cervical spine loads at the centers of mass of the head and vertebrae (C1 T1). The average peak loads and intervertebral motions were statistically compared (P < 0.05) throughout the cervical spine. RESULTS: Load and motion peaks generally increased with increasing impact acceleration. The average extension moment peaks at the lower cervical spine, reaching 40.7 Nm at C7-T1, significantly exceeded the moment peaks at the upper and middle cervical spine. The highest average axial tension peak of 276.9 N was observed at the head, significantly greater than at C4 through T1. The average axial compression peaks, reaching 223.2 N at C5, were significantly greater at C4 through T1, as compared to head-C1. The highest average posterior shear force peak of 269.5 N was observed at T1. CONCLUSION: During whiplash, the cervical spine is subjected to not only bending moments, but also axial and shear forces. These combined loads caused both intervertebral rotations and translations. PMID- 17114098 TI - Field data: distributions and costs of road-traffic fatalities in South Africa. AB - BACKGROUND: Road-traffic crashes and fatalities constitute major social and economic issues in South Africa. They are a major cause of morbidity and mortality comparable to HIV/AIDS, homicides, and some chronic diseases. METHODS: Road-traffic accident data for the year 2003 obtained from the Department of Transport, Pretoria, South Africa were used for this study. The valuation of the costs of road-traffic crashes and fatalities in South Africa is based on the gross output or human capital approach. RESULTS: 10,197 fatal road crashes and 12,353 fatalities were reported during the study period. More than 50% of the fatal road crashes and fatalities occurred in only three out of 11 provinces. The Northern Cape, which is the least populated province, had the highest fatal road crashes per 100,000 population and fatalities per 100,000 population. The number of road-traffic fatalities in the rural areas was 2.7 times that in the urban areas. The total costs of the road-traffic fatalities which was about R 8 billion (>US$ 1 billion) is about 0.6% of the country's nominal GDP for 2003. 60% of the cases and costs of road-traffic fatalities involved persons aged 20-39 years, although this age group is only 27% of the country's population. The rural areas accounted for 73% and the urban areas 27% of the total costs of fatal road traffic crashes. CONCLUSION: Those living in the rural areas of the South African society and those aged 20-49 years constitute high-risk groups of road-traffic crashes and fatalities. They are also responsible for most of the attendant costs of fatal crashes and fatalities in the country. PMID- 17114099 TI - Field data: a study on trend and prediction of fatal traffic injuries prevalence in Shanghai. AB - OBJECTIVE: The main objectives of the study are to analyze fatal traffic-injury trends in 1987-2003 in Shanghai and predict its prevalence in near future and provide scientific data for the local governmental decision on developing practical working methods on traffic-injury prevention and control. METHODS: In this study, epidemiological method and Grey dynamic model GM (1,1) were introduced to analyze and forecast traffic-injury mortality rates respectively. RESULTS: There was an apparent increasing trend of traffic-related injuries in Shanghai from 1987 to 2003 with the rate of growth in motorization. The average rates of annual increase are 3.59% in fatalities (from 7.78 per 100,000 population to 14.18 per 100,000 population) during the period. Pedestrians were the most common type of victims (29.6%), followed by bicyclists (25.1%), and motorcyclists (24.1%). Males accounted for the majority of all victims, over 69%. The population of high-school and lower high-school education level represented 66.4% victims of total road-traffic injuries. And if no special factors effect its development, the traffic fatalities would be up to 17.84 per 100,000 population in 2010, when calculating from equations we found and validated Y(t) = 359.90 x e0.027(t-1)-352.13, (t = 1, 2, ..., N) for Shanghai. CONCLUSION: Our data indicate the risk of fatal traffic injuries has increased in recent years and will go on growing in the near future in Shanghai. The findings showed that Grey dynamic model GM (1,1) is eligible on the prediction and can be a tool for injuries forecasting, implementing effective policies, programs, and interventions for reducing traffic injuries in the big cities. PMID- 17114100 TI - Study of biological aerated filters for the treatment of effluents from the citrus industry. AB - The primary objective of this research study was to test the applicability and optimize the design parameters of a system of biological aerated filters in order to obtain an optimal effluent from the citrus industry, which would allow its drainage into the municipal sewer system. Expanded clay was used as a support material. After experimenting with both countercurrent and cocurrent flows as well different aeration levels, it was found that cocurrent flow was more efficient. Backwashing was carried out on a daily basis. The results of our study showed that for an aeration of 10.47 Nm3/h/m2, the volumetric load should be less than 20 Kg. COD/m3/d in order to obtain an effluent with a maximum concentration of 600 mgCOD/L, and the hydraulic load should be less than 0.36 m/h. PMID- 17114101 TI - Toxicity of metal oxide nanoparticles in mammalian cells. AB - Along with existing and emerging use of nanoscale materials, growing concerns have arisen about their unintentional health and environmental impact. The objective of the ongoing study was to assess the toxicity profile of metal oxide nanoparticles proposed for use in industrial production methodology. Metal oxide nanoparticles used in this study included TiO2, ZnO, Fe3O4, Al2O3, and CrO3 with particle sizes ranging from 30 to 45 nm. Cellular morphology, mitochondrial function, membrane leakage of lactate dehydrogenase (LDH), permeability of plasma membrane, and apoptosis were assessed under controlled and exposed conditions (2 to 72 h of exposure). The microscopic studies demonstrated that nanoparticle exposed Neuro-2A cells (especially ZnO) at doses >100 microg/mL became abnormal in size, displaying cellular shrinkage, and detachment from the surface of flasks. Mitochondrial function decreased significantly in the cells exposed to ZnO at 50 to 100 microg/mL. However, Fe3O4, Al2O3, and TiO2 had no measurable effect on the cells until the concentrations reached greater than 200 microg/mL. LDH leakage significantly increased in the cells exposed to ZnO (50 to 100 microg/mL), while other nanoparticles tested displayed LDH leakage only at higher doses (>200 microg/mL). Flow cytometer tests showed that apoptosis took place in cells exposed to ZnO nanoparticles. More cells became necrotic as the concentrations increased. PMID- 17114102 TI - Comparative sensitivity of three tropical cladoceran species (Diaphanosoma brachyurum, Ceriodaphnia rigaudii and Moinodaphnia macleayi) to six chemicals. AB - The toxicities of six chemicals (cadmium chloride, potassium dichromate, sodium dodecyl sulfate, potassium chloride, Tritox X-100 and copper (II) sulphate) was determined for three tropical freshwater cladoceran, Moinodaphnia macleayi, Ceriodaphnia rigaudii and Diaphanosoma brachyurum. The data was subsequently used to compare the relative sensitivity of the three species. Relative sensitivities were determined by comparing the 48-hour LC50 values using an independent t-test and interspecies correlation. Potassium dichromate was the most toxic compound tested, while potassium chloride was the least toxic to all three species. The 48 h LC50 values for D. brachyurum ranged from 0.003 mg/L to 30.4 mg/L; M. macleayi LC50 values ranged from 0.003 mg/L to 30.1 mg/L whereas for C. rigaudii the values ranging between 0.002 mg/L to 21.1 mg/L. The LC50 values for C. rigaudii was significantly less (P < 0.05) than M. macleayi for five of the compounds tested, whereas for D. brachyurum it was significantly (P < 0.05) less than four of the compounds tested. The interspecies correlation also suggested that C. rigaudii and M. macleayi were more similar in sensitivity (R2 = 0.96) to each other than D. brachyurum (R2 = 0.91 with M. macleayi, and R2 = 0.87 with C. rigaudii). PMID- 17114103 TI - Sustainable approach for leachate treatment: electricity generation in microbial fuel cell. AB - Electricity generation from landfill leachate was examined by using both a dual chamber microbial fuel cell (MFC) and a single chamber MFC. Experimental results showed that the maximum power density of 2060.19 mW/m3 in the dual-chamber MFC and that of 6817.4 mW/m3 in the single chamber MFC were obtained. It was recognized that the difference in internal resistance for two MFC systems was the main reason for resulting in the difference of power generation. Power generation as function of chemical oxygen demand (COD) in single chamber MFC showed a Monod type relationship with Pmax of 5920.96 mW/m3 and Ks of 251.39 mg/L at an external resistance of 500 Omega. Cyclic voltammetry showed that electrons were directly transferred onto the anode by bacteria in biofilms, rather than self-produced mediators of bacteria in the solutions. At low COD concentration, electricity generation was limited by the anode due to kinetic limitation; while at high COD concentration, the cathode was shown to have more significant effects on the electricity generation than the anode. COD in leachate could be removed when it increases, mainly because oxygen diffused from the cathode was substantially reduced by aerobic or anoxic bacteria in the anode chamber, leading to the substrate loss. Removal of ammonium-nitrogen was not observed in the single chamber MFC. This novel technology provides an economical route for electricity energy recovery in leachate treatment. PMID- 17114104 TI - Determination of Pb(II) with a dithizone-modified carbon paste electrode. AB - A dithizone (DTZ) modified carbon paste electrode was developed for the sensitive and selective determination of Pb(II) using differential pulse anodic stripping voltammetry as well in batch as in FIA system. The analysis procedure is based on an open circuit accumulation step in a stirred sample solution. This was followed by a medium exchange to a clean solution and subsequently by a proper anodic stripping. The analytical performance was evaluated with respect to the quantity of modifier in the paste, accumulation time, background electrolyte, Pb(II) concentration and other variables. When the accumulation time applied was 5 minutes, linear calibration graphs were obtained in the concentration range 1 x 10(-7)-1 x 10(-5) M and 8 x 10(-8)-5 x 10(-6) M for batch and Flow Injection Analysis (FIA), respectively. The detection limits found were 8.65 x 10(-8) M in batch and 4.45 x 10(-8) M in FIA. A convenient and rapid renewal of electrode surface allows the use of a single modified electrode surface in multiple analytical determinations. Several coexisting metals ions such as Cd(II), Hg(II), Cu(II) and Zn(II) had no interference on the determination of Pb(II). The proposed method was applied in the determination of lead in soils located in the vicinity of metallurgic transformation industry. The results obtained were in accordance to the ones supplied by Atomic Absorption Spectroscopy (AAS). PMID- 17114105 TI - Characterization and source assessment of heavy metals and polycyclic aromatic hydrocarbons (PAHs) in sediments of the Fosu Lagoon, Ghana. AB - The first results ever obtained on polycyclic aromatic hydrocarbon (PAHs) concentrations in the Fosu lagoon surface sediments are presented together with corresponding heavy metal (Fe, Mn, Cd, Zn and Ni) concentrations. Samples collected on a monthly basis from November 2003-April 2004 (Heavy metals) and December 2003-January 2004 (PAHs) at 8 locations, representing different anthropogenic sources of contamination to the lagoon, were analyzed. Concentrations of Cd and Ni in the lagoon sediment suggest greater contamination to the lagoon from industrial activities in the vicinity of the lagoon; 50% of the sediment samples exceeded some established sediment Cd guidelines for the protection of aquatic lives. Then, 15 PAHs were detected among the sediments from the different locations and the compositional pattern in decreasing order was 3 ring > 5-ring > 4-ring > 6-ring > 2-ring PAH compounds. Sigma PAH concentrations in the sediment samples ranged from 254 to 558 mg/kg, with a mean of 359.4 mg/kg. Two distinct areas were identified to be a major source of anthropogenic load of both heavy metals and PAH; the mechanical shop in the northeastern sector of the lagoon is the main location for the input of Cd and it's associated PAH compounds (e.g., acenapthylene, acenaphthene, naphthalene and benzo[a]fluoranthene) and to a lesser extent Ni. Both combustion and petroleum sources may account for PAH loads from this area. The residential area in the northern sector is responsible for high loads of Mn and its associated PAH compounds (e.g., phenanthrene, benzo[a]pyrene and anthracene). These chemicals seem to enter the lagoon mainly by the combustion of especially wood or coal. PMID- 17114106 TI - Order of photocatalytic degradation as ranked by critical photonic times (CPTs) indicates the composition of organic dye mixtures: selectivity of hydroxyl radicals. AB - A simple critical photonic time (CPT) ranking method for the simultaneous determination of known organic dyes in the industrial wastewater of different concentrations was developed. A kinetic-measure, CPT theory, was developed to discriminate between the dyes in the textile effluent solution. The CPTs were calculated and ranked 1-4 from the smallest to the largest. The ranks indicate the order in which the individual dyes in the effluents were photocatalytically degraded. The described procedure allowed us to monitor the decolorization kinetics of the dyes in mixtures of different concentrations, when subjected to immobilized TiO2 photocatalysis in the presence of H2O2 from the start (H2O2FS). The outcome of hydroxyl radical (HO*) attack was consistently specific and regioselective for the individual dyes in the textile effluent. To explain the effect of HO*, the order of degradation of Acid Orange 52 < Acid Yellow 36 < Acid Red 17 < Acid Blue 45 < polyvinyl alcohol (PVA) was inferred. PMID- 17114107 TI - Total petroleum hydrocarbons and trace metals in street dusts from Tshwane Metropolitan area, South Africa. AB - This study reports the level of total petroleum hydrocarbons (TPHs) and trace metals in dusts from some petroleum handling facilities (gasoline stations), selected high traffic density roads and residential areas within the Tshwane Metropolitan area in South Africa. Total petroleum hydrocarbons in dust samples were extracted using soxhlet extraction and isolated gravimetrically after column cleanup, while total trace metals in dust samples were digested using mineral acid digestion and analyzed using atomic absorption spectrometry (AAS). The mean TPH level from the residential sites ranged from 206 +/- 20.3-300 +/- 36.4 microg g(-1) while those from gasoline stations and high traffic density roads varied between 562 +/- 43.9-340 +/- 62.6 microg g(-1) and 404 +/- 32.7-852 +/- 55.4 microg g(-1) respectively. Mean concentrations of analyzed trace metals at residential areas ranged from 0.04 +/- 0.02-0.07 +/- 0.03 microg g(-1); ND; 0.21 +/- 0.01-0.34 +/- 0.02 microg g(-1); 0.66 +/- 0.06-2.11 +/- 0.82 microg g(-1); 18.7 +/- 0.86-33.4 +/- 0.83 microg g(-1) and 0.07 +/- 0.04-0.23 +/- 0.01 microg g(-1) for Cd, Pb, Cu, Zn, Mn and Cr respectively. Mean levels from gasoline stations ranged between 0.05 +/- 0.02-0.17 +/- 0.07; 0.08 +/- 0.02-0.12 +/- 0.04; 0.21 +/- 0.04-4.34 +/- 0.60; 7.44 +/- 0.40-13.0 +/- 0.56; 23.36 +/- 4.37-131 +/- 10.5 and 0.05 +/- 0.02-0.17 +/- 0.05 microg g(-1) while those from high traffic density roads ranged from 0.07 +/- 0.04-0.10 +/- 0.04; ND-0.30 +/- 0.05; 0.67 +/- 0.05-3.80 +/- 0.82; 2.40 +/- 0.25-10.6 +/- 0.96; 19.8 +/- 1.03-108 +/- 4.44 and 0.06 +/- 0.02-0.12 +/- 0.04 microg g(-1) also in the above metal order. Contamination by TPH and trace metals both at gasoline stations and high traffic density roads is revealed by their significantly higher values compared to those of residential areas. PMID- 17114108 TI - Effect of surface tension reduction on VOC removal during surfactant-enhanced air sparging. AB - In this study, the effect of decreased surface tension of water on the removal efficiency of a volatile organic compound (VOC) during air sparing was evaluated using a laboratory-scale, two-dimensional physical model packed with sand and water containing dissolved toluene as a representative VOC. Two sets of air sparging experiments were performed: the first at a surface tension of 69 dyne/cm with no surfactant applied, and with a toluene concentration of 110 mg/L; the second at a surface tension of 50 dyne/cm due to 110 mg/L of anionic surfactant, and 99 mg/L of toluene. Under the experimental conditions used in this study, the sparging influence zone estimated at the lower surface tension was about 2.5 times that estimated at the high surface tension. Also the rate of toluene removal by air sparging was found to be much faster at the lower surface tension. The sparging time required for 50% removal of the initial mass of toluene was 16.8 hours at 50 dyne/cm, which was much smaller than 82.5 hours measured at 69 dyne/cm. The final mass recoveries at low and high surface tensions were 92.1% and 56.6%, respectively. Therefore it was concluded that air sparging at reduced surface tension (surfactant-enhanced air sparging) greatly improved the removal efficiency of VOCs from the porous medium compared to conventional air sparging with no applied surfactant. PMID- 17114109 TI - Influence of nitrate feeding on carbon dioxide fixation by microalgae. AB - In this study, the effects of nitrate feeding on microalgal growth and associated CO2 fixation were evaluated, as a strategy to enhance carbon fixation by increasing the duration of the exponential phase of cell growth in the batch operation of a photobioreactor. Two species of green algae, Chlorella and Scenedesmus, and two species of cyanobacteria, Microcystis ichthyoblabe and Microcystis aeruginosa, were used after adaptation to a 15% (v/v) CO2 environment. In the absence of nitrate feeding, nitrate concentrations declined rapidly and soon became a limiting factor. Nitrate feeding, administered in fed batch mode to maintain 15-20 ppm of NO3-N, allowed for an extension of the exponential growth phase by more than 3 days, as well as a higher cell density, which subsequently resulted in an increase in photoautotrophic carbon fixation. The increases in the carbon fixation rate were in the ranges of 56.1-56.6% for the green algae, and between 68.2-68.8% for the cyanobacteria. The results indicated that intermittent nitrate feeding was a viable strategy for the augmentation of fixation productivity, and may thus be effectively applied as a substitute for conventional medium change, which has traditionally been employed in order to prolong the active growth duration. PMID- 17114110 TI - Endocrine disrupting chemicals: human exposure and health risks. AB - Endocrine disrupting chemicals (EDCs) have been emphasized due to their threats in fertility, intelligence, and survival. For the last decade, many researchers have investigated EDC-health outcome. However, EDC responses in human were not clearly clarified through experimental and epidemiological data. Therefore, considering particular status of EDC endpoints, we suggest that one of the best ways to prevent unknown health risks from EDCs is to perform exposure monitoring or to do surveillance for EDC release into the environment. For this purpose, this review considers exposure status of EDCs, and EDC-related health risks, focusing on the mainly highlighted EDCs, such as dioxins/PCBs, DDT/DDE, bisphenol A, phthalates, alkylphenols, and phytoestrogens. We also reviewed tobacco, a mixed source of EDC-related endocrine disorders. PMID- 17114111 TI - Human health risk on environmental exposure to Bisphenol-A: a review. AB - Bisphenol-A (BPA), identified as an environmental hormone (i.e., endocrine disruptor), is an industrially important chemical that is being used as a primary raw material for the production of engineering plastics (e.g., polycarbonate/epoxy resins), food cans (i.e., lacquer coatings), and dental composites/sealants. From the ecotoxicology, human health and regulatory points of view, it is urgent to restrict the emissions and releases of the estrogenic chemical from the industrial processes and commercial products. This article reviews BPA in the current literature in terms of physiochemical properties, industrial/commercial uses, environmental distributions in the atmospheric/aquatic/terrestrial phases, possible human toxicity, and its exposure standards and limits. Emphasis is put on the most significant distribution in the aquatic environment, and occupational and non-occupational human exposures. Overall, it is strongly convinced that BPA is not a carcinogenic risk to humans, and is also rapidly glucuronidated and excreted through the route of urine. PMID- 17114112 TI - Occupational acrylonitrile exposure and lung cancer: a meta-analysis. AB - The present work summarizes the currently available published studies on lung cancer and occupational acrylonitrile exposure. Meta-analytic methods were used to estimate the overall risk. To adjust for the healthy worker effect, rate ratio estimates based on regression analyses and ratios of standard mortality ratios were aggregated. Overall effect estimates were 0.95 (95% CI 0.86 to 1.06) and 1.25 (95% CI 1.10 to 1.43) before and after adjustment for the healthy worker effect, respectively. Therefore, a 25% increase in lung cancer risk attributable to occupational acrylonitrile exposure is suggested. Possible contribution of smoking confounding the increased risk cannot be fully excluded. PMID- 17114114 TI - Tear glucose dynamics in diabetes mellitus. AB - This study compares tear glucose dynamic differences between 121 diabetic and nondiabetic subjects after the administration of a carbohydrate load. A quantitative chromatographic analysis of tear glucose was used and the values correlated to blood glucose values. Diabetic and nondiabetic tear glucose mean values were 0.35 +/- 0.04 mmol/L and 0.16 +/- 0.03 mmol/L, respectively. Significant differences were observed among the subject groups in both the tear and capillary blood glucose values. A correlation between tear glucose and capillary blood glucose was observed. The concentration of glucose in the tear fluid changes proportionately with respect to capillary blood glucose after a carbohydrate challenge. Although it is possible to determine the diabetic status of a subject using tear glucose values alone, in the clinical setting this may not prove to be practical due to technical limitations. PMID- 17114115 TI - Healing process at the flap edge in its influence in the development of corneal ectasia after LASIK. AB - PURPOSE: Post-laser in situ keratomileusis (LASIK) corneal ectasia is a progressive deformation of the gross corneal anatomy that occurs after surgery. However, this is a rare event even after deep lamellar keratoplasty. We hypothesize that the strength of the lamellar keratoplasty wound is derived from the sutures that enhance the wound edge healing response. This study compares, in a rabbit model, the stability of previously sutured and unsutured microkeratome flaps. METHODS: Unilateral 160-micro m-thick LASIK flaps using a mechanical microkeratome was performed in 20 rabbit eyes. Animals were then divided in two groups: In group A, the flap was left without sutures. In group B, the flap was sutured with 12 interrupted 10/0 nylon stitches that were removed after 3 weeks under general anesthesia. Six weeks after surgery, all rabbits had corneal topographies performed at their baseline intraocular pressure (IOP) (14 mmHg) and at two artificially increased pressures (25 and 45 mmHg) using an anterior chamber maintainer implanted in the inferior limbal area. The animals were humanely euthanized, and immunohistological analysis of the corneas was performed. RESULTS: A delta K1 value, which indicates the difference in the simulated keratometric value at baseline and the one measured at 25 mmHg, was calculated for all eyes. It showed a mean steepening effect of 2.74 D +/- 0.38 D in group A compared with 1.08 D +/- 0.27 D in group B (p < 0.05). Similarly, a delta K2 value, which indicates the difference in the simulated keratometric value at baseline and the one obtained at 45 mmHg, was registered. It showed a mean steepening effect of 3.02 D +/- 0.87 D in group A compared with 0.75 D +/- 0.44 D in group B (p < 0.05). Six weeks after surgery, the peripheral flap interface in group B consisted of 14.3% +/- 4.15% of positive monoclonal mouse anti-alpha smooth muscle actin (alpha-SMA) cells compared with 4.18 +/- 3.76% in group A (p < 0.05). CONCLUSIONS: The addition of sutures in the corneal flap after LASIK appears to reduce the amount of corneal steepening when the IOP is artificially increased up to 25 mmHg in this rabbit model. Our results suggest that an increase in the amount of myofibroblastas induced by the sutures may be responsible for this behavior. Corneal ectasia may be related to the clinically observed lack of corneal wound-healing at the edge of the flap that allows the cornea to bulge. By stimulating a stronger wound-healing response at the edge of the flap, the cornea may better resist steepening under increased IOP conditions and improve the long-term stability of LASIK surgery in borderline thin corneas. PMID- 17114116 TI - Filtering bleb evaluation with slit-lamp-adapted 1310-nm optical coherence tomography. AB - PURPOSE: Investigation of slit-lamp-adapted 1310-nm optical coherence tomography (OCT) as an in vivo imaging device in the postoperative course of glaucoma surgery. METHODS: Postoperative images of filtering blebs and deep sclerectomies and their healing processes were qualitatively evaluated with a slit-lamp-adapted anterior segment OCT (AS-OCT; Heidelberg Engineering, Heidelberg, Germany) in 28 patients. Ophthalmologic examinations included slit-lamp examination, applanation tonometry, and slit-lamp photography. The OCT scans were qualitatively correlated with the morphologic and functional outcome of the filtering bleb. RESULTS: 1310 nm OCT was able to demonstrate the internal structure and the dimensions of filtering blebs, as well as the scleral flap and the deep sclerectomy location including Descemet membrane. Functioning filtering blebs showed a low OCT signal, small fluid-filled cysts, superficial microcystic layer, and a slack internal texture. High internal reflectivity indicated an earlier scarring of the filtering bleb. Nonfunctioning filtering blebs delivered a high OCT signal, no or few cysts, and a dense internal texture. These different OCT patterns correlated with the clinical outcome. CONCLUSIONS: Slit-lamp-adapted 1310-nm OCT allowed the noncontact observation and documentation of the postoperative healing course of filtering blebs after glaucoma surgery. Internal structures of the filtering bleb and deep sclerectomies could be visualized. Functioning and dysfunctioning filtering blebs delivered different OCT pattern and correlated with the clinical outcome. This could be a new way to assess the postoperative healing process with the possibility of earlier intervention in cases of impending scarring. PMID- 17114117 TI - Immunolocalization of EMMPRIN (CD147) in the human eye and detection of soluble form of EMMPRIN in ocular fluids. AB - PURPOSE: To study the cellular distribution of extracellular matrix metalloproteinase inducer (EMMPRIN; CD147) in the human eye and the corneal and retinal pigment epithelium cell lines and its possible existence as a soluble protein in ocular fluids. METHODS: Immunohistochemistry was performed on human eyes and for cell cultures. Different EMMPRIN isoforms were analyzed by Western blotting in ocular fluids. RESULTS: EMMPRIN immunostaining could be detected in the corneal and conjunctival epithelium, the endothelium, and in the stromal keratocytes, the retinal pigment epithelium, several retinal layers and nerve fibers in the optic nerve head. Both cell lines deposit EMMPRIN on the cell membranes. Soluble EMMPRIN could also be detected in the tear fluid, aqueous humor, and vitreous samples in the form of multiple immunoreactive species. CONCLUSIONS: EMMPRIN is specifically expressed in the human eye only by certain tissue structures, thus suggesting specialized functions. The protein also exists naturally in soluble forms in ocular fluids representing presumably monomeric and multimeric forms, a notion that confirm and further extends its previously known role mainly as a transmembrane protein. These findings suggest that EMMPRIN can regulate not only cell surface functions in the human eye but also certain peri- and extracellular functions. PMID- 17114118 TI - Ultraviolet light-induced and green light-induced transient pupillary light reflex in mice. AB - PURPOSE: To study UV light-induced and green light-induced pupillary light reflex (PLR) in mice and to measure the illumination thresholds of rod-mediated and cone mediated PLR responses. METHODS: We measured dark-adapted transient PLR- in C57BL/6 mice elicited with ultraviolet and green light over an intensity range of 9 log units. To assist in isolating the responses mediated by rods and cones, we studied the PLRs in mouse models presenting pure cone and pure rod functions. We also characterized ERG signals in these mice under the same experimental conditions. RESULTS: The UV light-induced transient PLR has identical intensity response curves as green light-induced PLR in all the three mice strains. The threshold (5% PLR) of rod-driven PLR is 107 approximately 108 photons cm2 s-1, which is 1 approximately 2 log units lower than the dark-adapted ERG b-wave. The threshold of cone-driven PLR is approximately 1012.5 photons cm-2 s-1 and is similar to that of the cone ERG. CONCLUSIONS: We demonstrated that mice have PLR responses under UV stimulation. The cone-elicited PLRs have a threshold that is approximately 5 log units higher than that of rod-elicited PLR at both UV and green wavelengths. We observed a divergence between the spectra responses in PLR and ERG. However, the mechanism and implications of this phenomenon are yet to be identified. PMID- 17114119 TI - Effects of intravitreal dispase on vitreoretinal interface in rabbits. AB - OBJECTIVE: To explore the efficacy and safety of dispase on the production of posterior vitreous detachment (PVD) following intravitreal injection at various concentrations for different exposure times in the rabbit eyes. METHODS: Sixty four pigmented rabbits were assigned to two groups according to different combinations of concentrations of dispase and exposure times, namely group 1 (10, 5, or 1 U/ml for 30 min) and group 2 (0.25, 0.1, or 0.05 U/ml for one week). The control was established in both groups. One eye of each experimental rabbit was injected with dispase, and one eye of each control rabbit was injected with phosphate buffered saline. The presence of PVD and intraocular toxicity were observed by biomicroscope, macroexamination, and histology. RESULTS: None of the control eyes showed PVD. In group 1, PVD was present in 6 of 7 eyes in each sub group. In group 2, all 7 eyes of 0.25 U/ml or 0.1 U/ml and 6 of 7 eyes of 0.05 U/ml showed PVD. Intraocular toxicity including inflammation in anterior chamber, vitreous haze, epiretinal membrane, and retinal damage were found in the experimental eyes with relatively higher concentrations of dispase. CONCLUSIONS: Although dispase has a definite effect on the induction of PVD in rabbits, its toxicity to retina also alerts us to the application in clinic. The intravitreal injection of dispase of 1 U/ml for 30 min may be acceptable. PMID- 17114120 TI - Induction of endoplasmic reticulum stress in retinal pericytes by glucose deprivation. AB - Diabetic retinopathy is one of the major microvascular complications associated with diabetes mellitus, and the selective degeneration of retinal capillary pericytes is considered to be a hallmark of early retinopathy. Because glucose fluctuations commonly occur in diabetes, we hypothesized that these fluctuations will increase the endoplasmic reticulum (ER) stress and induce the unfolded protein response (UPR) in retinal pericytes. To study whether ER stress and the UPR can be induced in retinal pericytes, rat retinal capillary pericytes were cultured in different concentrations of glucose. Hypoglycemia but not hyperglycemia was found to activate UPR-specific enzymes in pericytes. Strong UPR activation leading to apoptosis was also observed when pericytes were cultured in glucose concentrations that were reduced from high to low or no glucose. These results indicate that induction of UPR is related not only to absolute concentrations but also to a shifting from higher to lower concentrations of glucose. PMID- 17114121 TI - Acute inflammation and loss of retinal architecture and function during experimental Bacillus endophthalmitis. AB - Rapid vision loss and explosive inflammation are devastating consequences of Bacillus endophthalmitis that have not been well defined. We therefore analyzed the evolution of intraocular inflammation and loss of retinal architecture and function during experimental Bacillus endophthalmitis. Mice were intravitreally injected with 100 CFU of B. cereus, and eyes were analyzed for bacterial growth, retinal function, architectural changes and retinal cellular stress, inflammatory cytokines, and infiltrating cells. Retinal electrophysiologic and structural changes began as early as 4 to 6 hr postinfection. Significant declines in retinal function paralleled the loss of retinal architecture. Glial fibrillary acidic protein (GFAP) was detected in retina, indicating potential stress. Polymorphonuclear leukocyte (PMN) infiltration into the vitreous began as early as 4 hr postinfection, coinciding with a significant increase in TNF-alpha in the eye. These results indicated that acute inflammation and detrimental architectural and electrophysiologic changes in the retina began earlier than once thought, suggesting that therapeutic intervention should be given at the earliest possible time to avoid vision loss during Bacillus endophthalmitis. PMID- 17114122 TI - Dendritic abnormalities in retinal ganglion cells of three-month diabetic rats. AB - PURPOSE: To determine the effect of three-month diabetes on the retinal ganglion cell (RGC) morphology and density. METHODS: Experimental diabetes was induced in Sprague-Dawley rats by intraperitoneal injection of streptozotocin (STZ). Retinas from three-month diabetic and age-matched control rats were harvested, and immunohistochemistry with monoclonal anti-Thy-1 antibody was carried out for calculating RGC density. Random RGC labeling with gene gun propelled lipophilic fluorescent dye, DiI, coated particles (DiOlistic method) was done for detailed RGC classification, dendritic field, and soma size measurement. RESULTS: The number of Thy1-labeled RGCs in the three-month diabetic rats was significantly reduced compared with that in the age-matched control. Obvious RGC morphology changes were observed, and the number of RGCs that could not be classified was significantly greater in the diabetic retinas. Among those well-classified RGCs, cells with enlarged dendritic fields were more frequently seen in the RGA group (401+/-86 um, n = 59, P < 0.001), but the soma sizes were unchanged from the controls (P > 0.05). For cells in the groups of RGB and RGC, no significant changes in the dendritic fields and soma sizes were found (P > 0.05). CONCLUSIONS: In the three-month STZ-induced diabetic rat, retinal ganglion cell loss is associated with morphology change. The surviving RGCs in the diabetic retina, especially those in RGA group, show significant dendritic field enlargement. This plasticity of the surviving RGC dendrites may represent a compensatory response to the overall loss of RGCs in diabetes. PMID- 17114123 TI - Erythrocyte membrane anionic content and urinary glycosaminoglycan excretion in type 1 diabetes: association with retinopathy. AB - BACKGROUND: In diabetic nephropathy, increased excretion of glycosaminoglycans and loss of basement membrane anionic charge had been documented to be related with diabetic microalbumuniria. There was no study that studied those two factors in clinical settings with a degree of diabetic retinopathy in type 1 diabetics. METHOD: Forty subjects (aged 27.3 +/- 6.3) with type 1 diabetes with different levels of diabetic retinopathy and 30 healthy subjects (aged 29.52 +/- 8.7) were included in the study. Subjects were first divided as patients without (R0) and with (R1) retinopathy. They then were further divided into two subgroups with the help of fundus angiography as diabetic lesions demonstrable with fluorescein angiography (R1A) and early diabetic retinopathy lesions on fundus examination (R1B). Erythrocyte anionic charge (EAC) was determined by the binding of cationic dye, alcian blue, and urinary glycosaminoglycan excretion and (UGAG) was determined spectrophotometrically. RESULTS: UGAG was increased (35.74 +/- 14.01 vs 21.25 +/- 6.19 micro g/mL, 95% confidence intervals [CI], 9.01-19.96, p = 0.02) and EAC (62.14 +/- 27.17 vs. 158.53 +/- 36.98 ng alcian blue 106 per 106 RBC, 95% [CI] -111.68-81.10, p = 0.0001) was decreased significantly in diabetic patients with respect to controls. As the grade of diabetic retinopathy increased, UGAG increased and EAC decreased within subgroups of diabetic patients (p < 0.005). UGAG positively correlated (r = 0.36 and p = 0.03) and EAC negatively (r = -0.695, p = 0.0001) correlated with diabetes duration. EAC and UGAG negatively correlated (r = -0.58 and p = 0.0001) with each other in type 1 diabetics. EAC (p = 0.007) and diabetes duration (p = 0.001) were found to be the two significant factors to have diabetic retinopathy in diabetics with logistic regression analysis. CONCLUSION: Although we need more prospective and larger studies to get a direct conclusion, we found that type 1 diabetic patients with less EAC and more UGAG are more likely to have diabetic retinopathy. PMID- 17114124 TI - Basic fibroblast growth factor suppresses retinal neuronal apoptosis in form deprivation myopia in chicks. AB - PURPOSE: Chorioretinal atrophy including retinal neuronal apoptosis occurred in chronic form-deprivation myopia induced by lid suture in chicks. We investigated whether exogenous basic fibroblast growth factor (bFGF) could simultaneously inhibit the excessive axial elongation and retinal neuronal loss in the myopic chick eyes. METHODS: Unilateral form deprivation was produced in neonatal male Hyline chicks by lid suture 24 hr after hatching. Ten microliters of solution containing 5 ng bFGF or PBS (vehicle) was injected into the vitreal chamber at 10 weeks of age, once every 3 days until week 12, when the animals were sacrificed. Ocular refraction and axial length were assessed by retinoscopy and calipers at the age of 12 weeks. Retinal apoptotic neurons and their caspase-3-like protease activity were analyzed by transmission electron microscopy, the terminal deoxynucleotidyl transferase-mediated biotin-deoxyuridine triphosphate nick-end labeling (TUNEL) staining, and a colorimetric method using Ac-DEVD-pNA as a substrate. The number of TUNEL-positive cells was counted to analyize the survival activity of bFGF on retina. RESULTS: After 12 weeks of lid suture, the control eyes were either emmetropic or hyperopic, whereas the deprived eyes became myopic with axial length increased. TUNEL-positive nuclei, condensed nuclear chromatin, and apoptotic bodies were observed in posterior retinal outer nuclear layer (ONL) and inner nuclear layer (INL) in the myopic chick eyes. Activity of retinal caspase-3-like protease in these eyes was elevated. In addition to ameliorating myopic ocular growth, intravitreal bFGF therapy significantly reduced the number of retinal apoptotic neurons and downregulated caspase-3 activity. CONCLUSIONS: Exogenous bFGF effectively ameliorates the excessive axial elongation and retinal neuron apoptosis in chronic form deprivation myopia in chicks. It is possible that bFGF will be a promising therapeutic agent for high human myopia. PMID- 17114126 TI - Occupational voice complaints and objective acoustic measurements-do they correlate? AB - To enable the development of appropriate diagnostics and treatment for occupational voice disorders, this study addresses connections between subjective voice complaints and objective observations. The subjects of this study were 24 female customer advisors, who mainly use the telephone during their working hours. During one working day, at four different times, speech samples covering 20 minutes of telephone conversation by the customer service advisors (CSAs) were recorded. In addition, the CSAs filled in a questionnaire (visual analogue scale) concerning their voice problems. To represent the vocal symptoms three variables were used: vocal fatigue, hoarseness and a general sum-variable. A 5-minute sample was taken from recordings for further analyses. This included fundamental frequency, sound pressure level, alpha ratio (the ratio between the spectral energy below and above 1000 Hz) and number of vocal fold vibrations. In the objective acoustic measurements, it was found that fundamental frequency (F0) rose significantly during the working day. Also the self-reported voice symptoms increased significantly during the working day. However, correlations between vocal symptoms and acoustic measures were not found. PMID- 17114127 TI - The role of F3 in the vocal expression of emotions. AB - The present study investigates the role of F3 in the perception of valence of emotional expressions by using a vowel [a:] with different F3 values: the original, one with F3 either lowered or raised by 30% in frequency, and one with F3 removed. The vowel [a:] was extracted from the simulated emotions, inverse filtered and manipulated. The resulting 12 synthesized samples were randomized and presented to 30 listeners who evaluated the valence (positiveness/negativeness) of the expressions. The vowel with raised F3 was perceived more often as positive than the sample with original (p = 0.063), lowered (p = 0.006) or removed F3 (p = 0.066). F3 may affect perception of valence if the signal has sufficient energy in high frequency range. PMID- 17114128 TI - Subglottal pressure and normalized amplitude quotient variation in classically trained baritone singers. AB - The subglottal pressure (Ps) and voice source characteristics of five professional baritone singers have been analyzed and the normalized amplitude quotient (NAQ), defined as the ratio between peak-to-peak pulse amplitude and the negative peak of the differentiated flow glottogram and normalized with respect to the period time, was used as an estimate of glottal adduction. The relationship between Ps and NAQ has been investigated in female subjects in two earlier studies. One of these revealed NAQ differences between both singing styles and phonation modes, and the other, based on register differences in female musical theatre singers, showed that NAQ differed between registers for the same Ps value. These studies thus suggest that NAQ and its variation with Ps represent a useful parameter in the analysis of voice source characteristics. The present study aims at increasing our knowledge of the NAQ parameter further by finding out how it varies with pitch and Ps in professional classically trained baritone singers, singing at high and low pitch (278 Hz and 139 Hz, respectively). Ten equally spaced Ps values were selected from three takes of the syllable [pae:], initiated at maximum vocal loudness and repeated with a continuously decreasing vocal loudness. The vowel sounds following the selected Ps peaks were inverse filtered. Data on peak-to-peak pulse amplitude, maximum flow declination rate and NAQ are presented. PMID- 17114129 TI - Voice problems experienced by Finnish comprehensive school teachers and realization of occupational health care. AB - A survey was conducted among teachers to explore 1) risk factors of voice disorders, 2) the number and nature of problems experienced, 3) the need for medical help, and 4) how occupational health care reacts to this need. Additionally, the knowledgeability of occupational health care physicians and the way treatment of voice disorders is currently organized was explored. Of the 181 teachers who responded, 42% reported voice symptoms occurring daily or weekly. Every tenth teacher also had vocal nodules, and 40% of those with recurring voice problems had been on sick-leave. The respondents suggested improvements in occupational health care: it should also cover prevention, waiting times need to be shortened and care chains established, and resources need to be allocated for voice therapy services. PMID- 17114130 TI - The intelligibility of tracheoesophageal speech, with an emphasis on the voiced voiceless distinction. AB - Total laryngectomy has far-reaching effects on vocal tract anatomy and physiology. The preferred method for restoring postlaryngectomy oral communication is prosthetic tracheoesophageal (TE) speech, which like laryngeal speech is pulmonary driven. TE speech quality is better than esophageal or electrolarynx speech quality, but still very deviant from laryngeal speech. For a better understanding of neoglottis physiology and for improving rehabilitation results, study of TE speech intelligibility remains important. Methods used were perceptual evaluation, acoustic analyses, and digital high-speed imaging. First results show large variations between speakers and especially difficulty in producing voiced-voiceless distinction. This paper discusses first results of our experiment. PMID- 17114131 TI - Spectral noise estimation in the evaluation of pathological voice. AB - A measure of the harmonics-to-noise ratio (HNR) in voice signals is used for assisting in the classification of voice pathologies. HNR estimation for voice signals has been investigated using time domain, cepstral, Fourier series and Fourier transform analyses. The present investigation focuses on methods that use a direct application of the Fourier transform to the signal. Three approaches to obtaining a HNR index from the power spectrum are reviewed. The present study uses synthetic voice signals to provide an unambiguous assessment of methods. The study highlights the fact that even though the indices derived from the power spectrum are useful in separating pathological and normal voice data sets, they provide only indirect information regarding the HNR of the glottal signal. PMID- 17114135 TI - What progress have we made with tinnitus? The Tonndorf Lecture 2005. AB - CONCLUSION: The field of tinnitus research is vibrant and active. Prospects for progress are high, but would be optimized by the growth of inter-disciplinary collaborations. OBJECTIVE: Tinnitus remains a source of urgent scientific investigation, and truly effective treatments continue to be elusive. The Tonndorf Lecture of the International Tinnitus Seminars represents an opportunity to reflect upon progress to date regarding tinnitus, and the actions needed to further that progress in future. METHOD: Progress regarding tinnitus mechanisms is reviewed, with particular regard to the conceptual distinction between ignition sites for tinnitus, and the physiological mechanisms that then promote the tinnitus through the central auditory pathway. The current status of both the Jastreboff neurophysiological model and the psychological model of tinnitus is reviewed. Some concerns regarding each model are raised, and the need for models that integrate the insights of both perspectives is identified as urgent. RESULTS: There are clear indications of progress in tinnitus, specifically regarding mechanisms, models and treatments. For knowledge to progress further, however, there is a pressing need for an inter-disciplinary approach to tinnitus, more involvement in teaching at a postgraduate level, and the development of experimental models of tinnitus that are both congruent with, and represent the complexity of, human experience of tinnitus. PMID- 17114136 TI - Cortical tonotopic map reorganization and its implications for treatment of tinnitus. AB - CONCLUSION: There appears to be a definite link between reorganization of the cortical tonotopic map and increased spontaneous firing rates. The results have implications for the reduction of noise-induced hearing loss and in the prevention of noise-induced tinnitus in humans. OBJECTIVES: To review animal and human studies related to neural correlates of tinnitus. Among those are increased spontaneous firing rate, enhanced neural synchrony, and reorganization of the cortical frequency-place (tonotopic) map. MATERIALS AND METHODS: To separate these issues one would want to have a situation where hearing loss is present but without reorganization of the cortical frequency-place map. For that purpose, noise-exposed cats were placed, immediately after the trauma and for at least 3 weeks, either in a quiet or in a high-frequency or low-frequency enriched acoustic environment. RESULTS: In exposed cats that were placed in the quiet environment there was an increase in spontaneous firing rate and synchrony of neurons in primary auditory cortex. In contrast, exposed cats placed in the high frequency-enriched acoustic environment did not show any significant difference in spontaneous firing rate or synchrony compared to the non-traumatized controls. PMID- 17114137 TI - Salicylate- and quinine-induced tinnitus and effects of memantine. AB - CONCLUSION: Memantine, an antiglutamatergic drug, has been proposed as a treatment for tinnitus. OBJECTIVES: The purpose of this study was to determine if memantine would prevent salicylate-induced tinnitus. Local field potentials were also recorded from auditory cortex to determine what effect salicylate, memantine, and the combination of both drugs would have on evoked potential amplitudes. MATERIALS AND METHODS: Schedule induced polydipsia-avoidance conditioning was used to identify the doses of salicylate or quinine that reliably induced tinnitus in rats. Rats were trained to lick for water during quiet intervals and avoid licking during sound intervals. RESULTS: Rats injected with saline or a low dose of sodium salicylate or quinine failed to develop tinnitus-like behaviors. However, high doses of salicylate (150-300 mg/kg/day) or quinine (100-150 mg/kg/day) greatly reduced licks-in-quiet, behavior consistent with the presence of tinnitus. Licks-in-quiet increased slightly when memantine (1.5 or 3 mg/kg/day) was co-administered with salicylate; however, the effect was not statistically significant or dose-dependent. These results indicate that memantine does not completely suppress salicylate-induced tinnitus. Cortical auditory evoked potential amplitude increased after salicylate treatment; co administration of memantine failed to block this salicylate-induced increase. PMID- 17114138 TI - Summary of evidence pointing to a role of the dorsal cochlear nucleus in the etiology of tinnitus. AB - Evidence has accumulated in the last decade that the dorsal cochlear nucleus (DCN) may be an important site in the etiology of tinnitus. This evidence comes from a combination of studies conducted in animals and humans. This paper will review the key findings, as follows. 1) Direct electrical stimulation of the DCN leads to changes in the loudness of tinnitus. This suggests that the loudness of tinnitus may be linked to changes in the level of neural activity in the DCN. 2) Exposure to tinnitus inducers, such as intense sound or cisplatin, causes neural activity in the DCN to become chronically elevated, a condition known as neuronal hyperactivity. 3) This hyperactivity is very similar to the activity that is evoked in the DCN by sound stimulation, suggesting that the hyperactivity represents a code that signals the presence of sound, even when there is no longer any sound stimulus. 4) Noise-induced hyperactivity in the DCN is correlated with tinnitus. Behavioral studies have demonstrated that animals exposed to the same intense sound that causes hyperactivity in the DCN develop tinnitus-like percepts. The correlation between the level of hyperactivity and the behavioral index of tinnitus was found to be statistically significant. 5) The DCN is a polysensory integration center, and electrophysiological studies have shown that both spontaneous activity and hyperactivity of neurons in the DCN can be modulated by stimulation of certain ipsilateral cranial nerves, such as the sensory branch of the trigeminal nerve. This ipsilateral modulation of DCN activity offers a plausible explanation of how tinnitus, when perceived on one side, can be modulated by certain manipulations of the head and neck on the side ipsilateral to the tinnitus, but rarely on the contralateral side. 6) The DCN exhibits various forms of neuronal plasticity that parallel the various forms of plasticity that characterize tinnitus. These findings collectively strengthen the view that the DCN may be a key structure that should be included as a target of anti-tinnitus treatment. PMID- 17114139 TI - Residual inhibition functions in relation to tinnitus spectra and auditory threshold shift. AB - CONCLUSIONS: Psychoacoustic functions relating the depth and duration of tinnitus suppression ('residual inhibition') to the center frequency of band-passed noise masking sounds appear to span the region of hearing loss, as do psychoacoustic measurements of the tinnitus spectrum. The results (1) suggest that cortical map reorganization induced by hearing loss is not the principal source of the tinnitus sensation and (2) provide a necessary baseline for optimizing residual inhibition in individual cases. OBJECTIVE: To measure residual inhibition functions and tinnitus spectra using sounds spanning the region of hearing loss. MATERIALS AND METHODS: Three subject-driven, computer-based tools were developed and applied to measure psychoacoustic properties of tinnitus and residual inhibition in 32 subjects with chronic tonal, ringing, or hissing tinnitus. Residual inhibition functions were measured with band-passed noise sounds varying in center frequency up to 12.0 kHz. RESULTS: The depth and duration of residual inhibition increased with the center frequency of the band-passed noise stimuli. Near-elimination of tinnitus for up to 45 s was reported by 8/24 (33%) subjects at center frequencies above 3 kHz (these cases distributed across tinnitus types). Tinnitus spectra covered the region of hearing loss with no preponderance of frequencies near the audiometric edge of normal hearing. PMID- 17114140 TI - New instrumentation for automated tinnitus psychoacoustic assessment. AB - CONCLUSION: Although tinnitus is a major health problem, techniques to quantify its perceptual aspects are not standardized. This study represents a key step in our efforts to develop clinical methodology to accurately and reliably quantify the sensation of tinnitus, using a uniform method for obtaining a battery of tinnitus measures. OBJECTIVES: The purpose of this study was to evaluate the performance of the automated system, which was redesigned to reduce time of testing and to add new testing capabilities. The primary difference in function was the use of a 'knob' device that enabled patient control of auditory stimuli. The new tests included assessment of minimum masking level (MML) and residual inhibition (RI). MATERIALS AND METHODS: As with previous iterations of the system, a computer program ran all testing and subjects read instructions and provided responses via a computer touch-screen. Three separate studies were conducted. Study 1 evaluated within- and between-session test-retest response reliability of tinnitus loudness matches (LMs) and pitch matches (PMs). Study 2 was conducted to evaluate differences in LMs and PMs between subjects with and without tinnitus - to obtain pilot data to assist in the development of a test for 'tinnitus malingering.' Study 3 evaluated the system's capability of obtaining MMLs and RI as well as the between-session reliability of these measures. RESULTS: Study 1 documented that the new system could obtain LMs and PMs within approximately 20 min, while maintaining clinically acceptable reliability. Study 2 revealed characteristic differences in LM and PM test results for individuals who did not experience tinnitus. Study 3 documented the system's ability to obtain measures of MML and RI that were reliable across sessions. PMID- 17114141 TI - The role of cognition in tinnitus. AB - CONCLUSIONS: The role of cognition in tinnitus is difficult to ignore. First, tinnitus is likely to disrupt cognitive functioning, and there are some indications that tinnitus patients have impaired capacity to perform certain cognitive tasks. Second, evidence is emerging that tinnitus patients show cognitive bias in the way they handle information. Such information processing style suggests either depressive functioning, or anxious vigilance, or both. Finally, self-report measures of tinnitus distress all require conscious recollection of how tinnitus is perceived and the consequences of tinnitus. Such reports necessitate cognitive capacity. OBJECTIVES: To review the literature on the interface between cognitive function and tinnitus, with special regard to the role of different levels of information processing. MATERIALS AND METHODS: A selective systematic literature search was conducted using the search engines of Medline and Psychological Abstracts, and by hand search of conference proceedings. RESULTS: There are yet relatively few published studies on cognitive functioning in tinnitus patients. Most research has been conducted by a few separate research groups. However, the available studies clearly implicate an important role of cognitive processes at different levels from basic cognitive function to more conscious appraisal of the consequences of tinnitus. Finally, a tentative model of the road from tinnitus generation to annoyance via cognitive function is suggested. PMID- 17114142 TI - Considerations for the design of clinical trials for tinnitus. AB - CONCLUSION: There are many possible control conditions to consider in designing research on tinnitus treatments. Some of the counseling procedures involve more than simply 'talking' or providing information, and it is important to make this distinction. Several good handicap scales are available, but we believe that 100 point interval scales have some superior attributes. Both primary and secondary measures of benefit should be used. Open trials have some merit, but should only be used cautiously. Several recent guidelines have been suggested for improving the design and reporting of clinical trials. OBJECTIVES: This paper reviews some basic considerations in the design of research to evaluate tinnitus treatments, particularly counseling and sound therapies. METHODS: We have reviewed some of the basic issues, referenced some relevant work, and provided some data supporting some of our assertions. RESULTS: We provide some recommendations for consideration for the design of clinical trials. PMID- 17114143 TI - Amygdalohippocampal involvement in tinnitus and auditory memory. AB - CONCLUSION: The preliminary results suggest that in chronic unilateral tinnitus the contralateral amygdalohippocampal complex does seem to be involved in tinnitus perception of pure tones. OBJECTIVES: Functional neuroimaging studies have revealed that the hippocampus and amygdala are involved in tinnitus perception. The amygdala and hippocampus are supplied by the anterior choroidal artery. Selective amobarbital injections in the anterior choroidal artery result in a non-functional amygdalohippocampal area for 10 min. The aim of this study was to assess the influence of this procedure on tinnitus perception. PATIENTS AND METHODS: Amobarbital (80 mg in total) was injected selectively in two sessions in the left and right anterior choroidal artery in six male patients with tinnitus: four with unilateral tinnitus, two with bilateral tinnitus. Of the patients with unilateral tinnitus, three had right-sided tinnitus and one had left-sided tinnitus. The average age was 57.3 years (range 43-69). Average tinnitus duration was 5.3 years (range 1-10). The differences in visual analogue scale (VAS) scores before and after the amytal tests were analysed. RESULTS: Amytal injection ipsilateral to the side where the tinnitus was perceived resulted in a maximum of 30% tinnitus suppression, whereas amytal injection contralateral to the tinnitus side yielded a 60-70% tinnitus suppression in three patients with unilateral chronic tinnitus (>4 years). Only pure tone tinnitus was suppressed, white noise was not. Two patients with bilateral tinnitus had no suppression, irrespective of the tinnitus type. A third patient without clinical tinnitus suppression had tinnitus of more recent origin (1.5 years). PMID- 17114144 TI - Acoustic shock injury (ASI). AB - CONCLUSION: The potential severity and persistence of ASI symptoms has significant clinical and medico-legal implications. With the rapid growth of call centres around the world, professionals providing tinnitus and hyperacusis therapy are increasingly likely to encounter some or all of the cluster of ASI symptoms in their clients. BACKGROUND: Acoustic shock injury (ASI), occurring as a result of exposure to a sudden unexpected loud sound, has been observed to cause a specific and consistent pattern of neurophysiological and psychological symptoms. These include aural pain, tinnitus, hyperacusis/phonophobia, vertigo and other unusual symptoms such as numbness or burning sensations around the ear. A range of emotional reactions including trauma, anxiety and depression can develop. Call centre staff using a telephone headset or handset are vulnerable to ASI because of the increased likelihood of exposure, close to their ear(s), of sudden unexpected loud sounds randomly transmitted via the telephone line. DISCUSSION: This paper presents an overview of a study of 103 people exposed to 123 acoustic incidents, and of the proposed neurophysiological mechanism of ASI, in particular tonic tensor tympani syndrome (TTTS). An understanding of TTTS has the potential to provide insight into the neurophysiological and psychological development of tinnitus and hyperacusis and the association with high levels of emotional trauma and anxiety. REHABILITATION: ASI rehabilitation is discussed. PMID- 17114145 TI - Use of bedside sound generators by patients with tinnitus-related sleeping difficulty: which sounds are preferred and why? AB - CONCLUSIONS: Most tinnitus patients who have difficulty sleeping experience some improvement in sleep after short-term use of bedside sound generators (BSSGs), although this study does not allow conclusions to be drawn as to how much other factors contribute. Many patients seem to find BSSGs helpful in reducing autonomic arousal. Further research is needed, but these findings raise the possibility that the emotional effects of sound enrichment have an important role to play in improving sleep among tinnitus patients. OBJECTIVES: This study investigated which sounds out of the options available on BSSGs are commonly chosen by patients and the reasons behind these choices. It also aimed to provide an indication as to whether BSSGs improve sleep quality in the short term. PATIENTS AND METHODS: A consecutive series of 39 tinnitus clinic patients who made a subjective complaint of sleep disturbance took part in the study. All participants were given a Naturecare BSSG to use at night. The Pittsburgh Sleep Quality Index (PSQI) and a semi-structured interview were used as outcome measures. RESULTS: Among the 35 participants who attended for follow-up there was a significant improvement in PSQI scores (p=0.001). 'Brook' and 'birds' were the most popular sounds, while 'white noise' proved the least popular. Most BSSG users listened to one sound only and most said that they chose their sound because of a pleasant emotional effect. A minority gave the quality of sound or its perceived effect on tinnitus as a reason for their choice. PMID- 17114146 TI - Clinical trial to compare tinnitus masking and tinnitus retraining therapy. AB - CONCLUSION: Both tinnitus masking (TM) and tinnitus retraining therapy (TRT) can be effective therapies for amelioration of tinnitus. TM may be more effective for patients in the short term, but with continued treatment TRT may produce the greatest effects. OBJECTIVES: Although TM and TRT have been used for many years, research has not documented definitively the efficacy of these methods. The present study was a controlled clinical trial to prospectively evaluate the clinical efficacy of these two methods for US military veterans with severe tinnitus. SUBJECTS AND METHODS: Over 800 veterans were screened to ensure that enrolled patients had tinnitus of sufficient severity to justify 18 months of individualized treatment. Qualifying patients (n=123) were placed quasi-randomly (alternating placement) into treatment with either TM or TRT. Treatment was administered at 0, 3, 6, 12, and 18 months. Outcomes of treatment were evaluated primarily using three self-administered tinnitus questionnaires (Tinnitus Handicap Inventory, Tinnitus Handicap Questionnaire, Tinnitus Severity Index). RESULTS: Findings are presented from the three written questionnaires with respect to three categories of patients: describing tinnitus as a 'moderate,' 'big,' and 'very big' problem at baseline. Based on effect sizes, both groups showed considerable improvement overall. In general, TM effects remained fairly constant over time while TRT effects improved incrementally. For the patients with a 'moderate' and 'big' problem, TM provided the greatest benefit at 3 and 6 months; benefit to these TRT patients was slightly greater at 12 months, and much greater at 18 months. For patients with a 'very big' problem, TM provided the greatest benefit at 3 months. For these latter patients, results were about the same between groups at 6 months, and improvement for TRT was much greater at 12 months, with further gains at 18 months. PMID- 17114147 TI - 15-year prospective follow-up study of behavioral therapy in a large sample of inpatients with chronic tinnitus. AB - CONCLUSION: The results of this study are in accordance with the assumption that cognitive-oriented therapy enabling the patient to live with tinnitus is of primary importance to enhance quality of life. BACKGROUND AND OBJECTIVES: This study describes the success of an integrative behavioral-medicine inpatient treatment for complex chronic tinnitus and presents its long-term effects. In 1987 we developed and evaluated a new treatment concept of psychological treatment of complex chronic tinnitus based on international experience and results. To evaluate the influence, effects and individual results of the specific therapy we analyzed the data of 434 consecutively treated patients. To investigate the long-term effects of the treatment, we contacted the patients 15 years after discharge from the hospital. PATIENTS AND METHODS: We used the tinnitus questionnaire (TQ) and visual analog scales (VAS) for specific tinnitus variables (loudness, discomfort, control of tinnitus, stress, general mood). The German version of the Derogatis psychopathology checklist (SCL-90-R) was used to analyze the impact of additional symptoms (depression, anxiety, introversion, etc.). RESULTS: Compared with a control group (patients on a waiting list) significant and clinically relevant effects were found. At the outcome, there were significant improvements in almost all parameters measured. For evaluation of the long-term effect we succeeded in contacting 312 of 434 former patients. Data were assessed using the same questionnaires that had been employed at the first contact. In all, 271 patients (86%) returned the questionnaires. Data for 244 cases (mean age 63 years; 79 females, 165 males) were complete enough to be used for data analysis. The results of the follow-up were as unexpected as clear: 15 years after conclusion of the treatment, the improvements of the tinnitus parameters and additional symptoms were stable when compared with the end of therapy. PMID- 17114148 TI - Auditory discrimination therapy (ADT) for tinnitus managment: preliminary results. AB - CONCLUSION: This clinical assay has demonstrated the efficacy of auditory discrimination therapy (ADT) in tinnitus management compared with a waiting-list group. In all, 43% of the ADT patients improved their tinnitus, and its intensity together with its handicap were statistically decreased (EMB rating: B-2). OBJECTIVE: To describe the effect of sound discrimination training on tinnitus. ADT designs a procedure to increase the cortical representation of trained frequencies (damaged cochlear areas with a secondary reduction of cortical stimulation) and to shrink the neighbouring over-represented ones (corresponding to tinnitus pitch). STUDY DESIGN: This prospective descriptive study included 14 patients with high frequency matched tinnitus. Tinnitus severity was measured according to a visual analogue scale (VAS) and the Tinnitus Handicap Inventory (THI). Patients performed a 10-min auditory discrimination task twice a day for 1 month. Discontinuous 8 kHz pure tones were randomly mixed with 500 ms 'white noise' sounds through a MP3 system. ADT group results were compared with a waiting-list group (n=21). RESULTS: In all, 43% of our patients had improvement in their tinnitus. A significant improvement in VAS (p=0.004) and THI mean scores was achieved (p=0.038). Statistical differences between ADT and the waiting-list group have been proved, considering patients' self-evaluations (p=0.043) and VAS scores (p=0.004). A non-significant reduction of THI was achieved (p=0.113). PMID- 17114149 TI - The impact of auditory cortex activity on characterizing and treating patients with chronic tinnitus--first results from a PET study. AB - CONCLUSION: Unilaterally increased metabolic activity within the primary auditory cortex (PAC) represents a robust finding in tinnitus patients. Targeting these hyperactive areas with image-guided low frequency repetitive transcranial magnetic stimulation (rTMS) results in subjective tinnitus reduction. More pronounced activation of the PAC predicted higher resistance to rTMS. OBJECTIVES: [18F]deoxyglucose (FDG)-positron emission tomography (PET) was used to assess metabolic activity within the central auditory system in tinnitus. The study investigated whether patterns of neuronal activity correlate with clinical features or may be used for the prediction of treatment outcome. PATIENTS AND METHODS: Twenty patients with chronic tinnitus underwent PET imaging followed by low frequency rTMS treatment. Neuroimaging data were compared with clinical parameters and treatment outcome. RESULTS: PET data demonstrated an asymmetric activation of the central auditory system. Seventeen patients revealed increased activity of the primary auditory cortex on the left side, three on the right side. The extent of hypermetabolic activity prior to treatment correlated significantly with tinnitus reduction after rTMS, but not with clinical characteristics such as tinnitus severity, tinnitus laterality or tinnitus duration. PMID- 17114150 TI - Strategies of the Tinnitus Research Consortium. AB - CONCLUSIONS: Research supported by private philanthropy complements governmental support of research, and its organization can undertake analysis of the whole field, identify the initial steps required to advance the field, draw attention to the intellectual challenge of a field such as tinnitus, recruit scientists to a neglected area, direct support to the most promising research approaches and opportunities, and dedicate support to those endeavors. OBJECTIVES: The efforts of the Tinnitus Research Consortium (TRC) are to accelerate progress in basic and clinical research on tinnitus. METHODS: The TRC analyzes the field of tinnitus research, brainstorms for new research approaches to tinnitus and provides guidance to the scientific community through requests for applications (RFAs) on promising research approaches and opportunities. The analysis of the research field has focused on the validity of animal models of tinnitus, the need for a standardized outcome measure in clinical trials, and the control of confounding variables in basic and clinical studies of the mechanisms of and site(s) associated with tinnitus. RESULTS: The TRC judged that the existing animal models were worthy of further study and refinement and that additional animal models should be developed. In response to an RFA, a project for the development and validation of the new Tinnitus Functional Index was initiated. A confounding aspect of experimental induction of tinnitus is the occurrence of hearing loss. The experimental manipulation causing the hearing loss may or may not cause tinnitus. Segregation of the correlates of tinnitus from the consequences of hearing loss requires the inclusion of a second control group: animals that have been subjected to the manipulation for the induction of tinnitus and have a hearing loss but fail to exhibit signs of tinnitus. Comparison of normal animals, animals with hearing loss without tinnitus and animals with hearing loss and tinnitus greatly enhances the value of the research. Clinical research on the mechanisms and sites of tinnitus is also confounded by this problem, and the solution is to include controls matched for hearing loss without tinnitus. PMID- 17114151 TI - The use of flupirtine in treatment of tinnitus. AB - CONCLUSION: Flupirtine, a functional NMDA antagonist, does not seem to be efficacious in the treatment of tinnitus. OBJECTIVES: The purpose of this study was to investigate whether flupirtine has any beneficial effect on tinnitus perception. PATIENTS AND METHODS: Twenty-four patients were selected (6 female and 18 male patients) with continuous subjective tinnitus. Eight patients suffered left-sided tinnitus, 4 right-sided tinnitus and 12 bilateral tinnitus. We assessed the burden of the tinnitus by loudness visual analogue scale (VAS) and tinnitus questionnaire (TQ) according to Hallam et al., and Hiller and Goebel. All patients were treated with a 2 x 100 mg daily dosage of oral flupirtine for 3 weeks in an open prospective design. RESULTS: There was no statistical effect on VAS and TQ of the treatment with flupirtine. Only one patient (4.2%) experienced a positive effect on the tinnitus but discontinued the treatment because of amnesia and concentration disorders. PMID- 17114152 TI - Effects of repetitive transcranial magnetic stimulation (rTMS) on chronic tinnitus. AB - CONCLUSIONS: Preliminary results indicate that repetitive transcranial magnetic stimulation (rTMS) can suppress tinnitus for some people. However, several procedural questions need to be addressed before the results of TMS studies can be interpreted or applied. For example, the placebo effect might be a significant factor because it is easy for patients to distinguish between real and sham stimulation. OBJECTIVES: This study was undertaken to determine the following: Can rTMS reduce patients' perception of chronic tinnitus? Is ipsilateral or contralateral stimulation most effective at reducing patients' perception of tinnitus? What is the extent and duration of the change in tinnitus following rTMS? PATIENTS AND METHODS: Fifteen subjects rated the loudness of their tinnitus on a 1-10 scale (1 = very quiet, 10 = very loud) before and after sham or real TMS. Participants wore foam ear plugs during the following procedures. After the patient's motor threshold was established, a figure-of-eight stimulating coil was positioned over the temporal region of the head at a location that corresponds to International 10-20 electrode position T3 (left) or T4 (right). 'Sham' rTMS was then delivered to this region of the head, first on the ipsilateral side, then on the contralateral side from where participants perceived tinnitus. Sham TMS consisted of an audio recording of actual TMS stimulus sounds. Actual rTMS was next delivered to each side of the head, starting with the side ipsilateral to tinnitus perception. Stimulation intensity was 100% of the resting motor threshold. A train of 30 pulses at 10 Hz was delivered every minute for 5 min. RESULTS: Two subjects reported reductions in tinnitus loudness following sham stimulation. Actual TMS resulted in partial suppression of tinnitus for six subjects. The amount of suppression ranged from 19% to 86% (average 50%). The durations of tinnitus suppression for each of the six subjects were: 20 min, 30 min, 45 min, 60 min, and 1 and 4 days, respectively. PMID- 17114153 TI - Repetitive transcranial magnetic stimulation and chronic tinnitus. AB - CONCLUSION: There is a good theoretical basis and early research evidence suggesting that transcranial magnetic stimulation (TMS) may have treatment potential in tinnitus. Further studies with larger sample sizes and additional assessment of neurobiological effects are necessary. OBJECTIVES: Tinnitus is a common and often severely disabling disorder for which there is no satisfactory treatment. TMS is a new, non-invasive method of modifying the excitability of the cerebral cortex, which has proven effective in auditory hallucinations and other disorders. Some early studies have been published in which TMS has been used in the treatment of tinnitus. The objective of this paper is to examine the literature and consider the potential for TMS as a therapy in tinnitus. METHODS: A thorough search of the tinnitus and TMS literature was conducted, and all available relevant material was examined. RESULTS: Tinnitus is common, with a prevalence of 8.2% in subjects aged 50 years and over, and may be associated with great distress (tinnitus sufferers). There are no effective treatments. Tinnitus is frequently associated with deafness, and may be the result of a pathological plasticity process. Neuroimaging studies demonstrate increased activity within the central auditory system. TMS is a non-invasive method of modulating excitability in cerebral cortex. It uses electromagnetic principles and has been successfully employed in the treatment of other conditions associated with increased activity of the cerebral cortex. Meanwhile, a growing number of studies suggest that repetitive TMS may be effective in the treatment of chronic tinnitus. PMID- 17114154 TI - Job-related social networks and communication technology. AB - In order to examine the social networks of individuals who rely on AAC, 38 adults who used AAC provided information about how they developed and maintained job related social networks and how communication technologies helped them to do so. The respondents met new people who might become part of their job-related networks during social events with family or friends, as well as at meetings, conferences, and workshops. They also frequently used generic communication technologies such as the phone, email, computer, and the Internet to maintain contact with people in their social networks. Findings suggest that land-line telephones, email, and the Internet were used by the largest percentage of respondents, with email and the Internet used most frequently to contact the largest number of people. Qualitative data provided information about the use of technology for enhancing participants' job-related networks and suggestions for new or improved technologies. PMID- 17114155 TI - Exploring visual-graphic symbol acquisition by pre-school age children with developmental and language delays. AB - The process of language acquisition requires an individual to organize the world through a system of symbols and referents. For children with severe intellectual disabilities and language delays, the ability to link a symbol to its referent may be a difficult task. In addition to the intervention strategy, issues such as the visual complexity and iconicity of a symbol arise when deciding what to select as a medium to teach language. This study explored the ability of four pre school age children with developmental and language delays to acquire the meanings of Blissymbols and lexigrams using an observational experiential language intervention. In production, all four of the participants demonstrated symbol-referent relationships, while in comprehension, three of the four participants demonstrated at least emerging symbol-referent relationships. Although the number of symbols learned across participants varied, there were no differences between the learning of arbitrary and comparatively iconic symbols. The participants' comprehension skills appeared to influence their performance. PMID- 17114156 TI - What happens to reading between first and third grade? Implications for students who use AAC. AB - School-age students who use AAC need access to communication, reading, and writing tools that can support them to actively engage in literacy learning. They also require access to core literacy learning opportunities across grade levels that foster development of conventional literacy skills. The importance of the acquisition of conventional literacy skills for students who use AAC cannot be overemphasized. And yet, one of the critical challenges in supporting the literacy learning of students who use AAC has been a lack of knowledge about literacy curricula and supports to literacy learning for these students. Most students who use AAC do not become conventionally literate and few of those who do achieve literacy skills beyond the second grade level. This article will provide an overview of the most frequent reading instructional activities in first and third grade classrooms. To better understand the foundational experiences important to literacy learning, the results of a survey project that examined the reading activities of general education students and teachers during primary grade instruction are presented, and critical shifts in instruction that occurred between first and third grade are highlighted. The primary instructional focus of core reading activities is also examined, along with adaptations for students who use AAC. PMID- 17114157 TI - Augmentative and alternative communication use and acceptance by adults with traumatic brain injury. AB - The purpose of this study was to document augmentative and alternative communication (AAC) acceptance and use patterns of 25 adults with traumatic brain injuries (TBI) who used either high- or low-tech AAC devices or strategies at some point during their recovery. Specifically, the purposes were to (a) document acceptance of AAC system recommendations, (b) identify AAC use patterns by persons who accepted the recommendation and for whom AAC intervention was implemented, (c) identify AAC access patterns for message formulation and encoding, and (d) document the kind of communicative functions that different AAC strategies supported. Information was gathered via a questionnaire from speech language pathologists who provided AAC assessments and interventions at six different sites. The speech-language pathologists provided information about individuals with TBI from their clinics for whom they had recommended AAC. Results revealed that these adults generally accepted both high- and low-tech AAC recommendations and used their AAC systems for extended periods of time. Most utilized letter-by-letter message formulation strategies. When AAC technology was abandoned, it was usually a reflection of a loss of facilitator support rather than a rejection of the technology. PMID- 17114158 TI - Identifying patterns of communicative behaviors in girls with Rett syndrome. AB - The purpose of this study was to investigate behaviors of girls with Rett syndrome under different conditions and to determine if there were patterns of functional communicative behaviors. Six communication protocols portraying familiar and non-familiar activities were created to assist in identifying specified behaviors. The protocols incorporated activities and pre-scheduled interruptions designed to enhance communicative behaviors. Eight girls in the third and fourth stages of Rett syndrome were videotaped during the protocols. Results indicated that the girls' alternating eye gazing increased and persistence decreased when activities were interrupted. Additional behaviors varied by participant across different protocols. Several patterns of effective and ineffective communication were observed. Implications for practice and future research are discussed. PMID- 17114159 TI - Reducing the risk of sexual abuse for people who use augmentative and alternative communication. AB - To date little attention has been focused on the sexual abuse experiences of people who use augmentative and alternative communication (AAC) and on addressing ways to reduce their risk for this type of abuse. This paper describes the results of a 3-year project that aimed to: (a) learn about the sexual abuse experiences of people who use AAC; (b) provide educational forums and resources on topics relating to sexual abuse for adults who use AAC; (c) define implications in risk reduction for various community service workers who support people who use AAC (e.g., attendant service providers, abuse counselors, sexual health educators, police, victim assistance services, legal professionals, and health care professionals); and (d) make recommendations to parents, educators, service providers, and consumer advocacy organizations about their roles in reducing the risk of abuse for youth and adults who use AAC. The findings suggest that the majority of participants in this project have experienced a range of abuses including sexual abuse, lack information about healthy and abusive relationships, have no way of communicating about sexuality and abuse, and lack supports in their personal lives and from within the community-at-large that are necessary to cope with relationship difficulties and specifically problems associated with abuse and justice system services. These findings and implications are shared with the intent of highlighting the need for more research and attention to the issue of abuse prevention for people who use AAC. PMID- 17114160 TI - The phonological awareness abilities of children with cerebral palsy who do not speak. AB - To investigate the importance of the connection between being able to speak and the emergence of phonological awareness abilities, the performance of children with cerebral palsy (five speakers and six non-speakers) was assessed at syllable, onset-rime, and phoneme levels. The children were matched with control groups of children for non-verbal intelligence. No group differences were found for the identification of syllables, reading non-words, or judging spoken rhyme. The children with cerebral palsy who could speak, however, performed better than the children with cerebral palsy who could not speak and the control group of children without disabilities, judging written words for rhyme. The children with cerebral palsy who could not speak performed poorly in comparison to those who could speak (but not the control group of children) when segmenting syllables and on the phoneme manipulation task. The findings suggest that non-speaking children with cerebral palsy have phonological awareness performance that varies according to the mental processing demands of the task. The ability to speak facilitates performance when phonological awareness tasks (written rhyme judgment, syllable segmentation, and phoneme manipulation) require the use of an articulatory loop. PMID- 17114161 TI - Concomitant use of the matrix strategy and the mand-model procedure in teaching graphic symbol combinations. AB - Matrix strategies employing parts of speech arranged in systematic language matrices and milieu language teaching strategies have been successfully used to teach word combining skills to children who have cognitive disabilities and some functional speech. The present study investigated the acquisition and generalized production of two-term semantic relationships in a new population using new types of symbols. Three children with cognitive disabilities and little or no functional speech were taught to combine graphic symbols. The matrix strategy and the mand-model procedure were used concomitantly as intervention procedures. A multiple probe design across sets of action-object combinations with generalization probes of untrained combinations was used to teach the production of graphic symbol combinations. Results indicated that two of the three children learned the early syntactic-semantic rule of combining action-object symbols and demonstrated generalization to untrained action-object combinations and generalization across trainers. The results and future directions for research are discussed. PMID- 17114162 TI - AAC and community partnerships: the participation path to community inclusion. AB - The Life Needs Model (LNM) of service delivery emphasizes the importance of community participation in children's development and quality of life. This article is a case illustration of two community partnership programs based on the LNM. StoryTime and Dress Up and Drama provide participation opportunities in the community for children who use Augmentative and Alternative Communication (AAC). The authors describe the development and implementation of both programs. Regardless of level of functioning, each child who participated in the programs did so by communicating, interacting socially, and controlling the environment. Multiple benefits for children, parents/caregivers, community partners, and AAC clinicians are highlighted. The authors hope that this article will assist others in developing partnerships and implementing inclusive practices within their communities. PMID- 17114163 TI - Accessible information for people with complex communication needs. AB - Information can be empowering if it is accessible. While a number of known information access barriers have been reported for the broader group of people with disabilities, specific information issues for people with complex communication needs have not been previously reported. In this consumer-focused study, the accessibility of information design and dissemination practices were discussed by 17 people with complex communication needs; by eight parents, advocates, therapists, and agency representatives in focus groups; and by seven individuals in individual interviews. Participants explored issues and made recommendations for content, including language, visual and audio supports; print accessibility; physical access; and human support for information access. Consumer-generated accessibility guidelines were an outcome of this study. PMID- 17114164 TI - Purposes of AAC device use for persons with ALS as reported by caregivers. AB - Thirty-four informal caregivers who support 26 persons with ALS reported on AAC technology use. Each caregiver completed the Communication Device Use Checklist, a survey tool developed for this study based on Light's (1988) classification of the purposes of social interaction (Augmentative and Alternative Communication, 4, 66-82). The checklist includes 17 purposes of communication and asks participants to judge importance, mode, and frequency of use for each purpose. Results show that the three communication purposes used most frequently and valued as important by caregivers involve regulating the behavior of others for basic needs and wants (getting needs met; giving instructions or directions to others; and clarifying needs). Consistent reports of use and frequency for the purposes of staying connected (social closeness) and discussing important issues (information transfer) indicate that AAC technology can assist the dyad in maintaining previous relationships. The face-to-face spontaneous conversation mode is used most frequently, despite the slow rate of production, the lack of permanence, and the demands on conversational partners during message generation. Clinical and research implications are discussed. PMID- 17114165 TI - Use of safe-laser access technology to increase head movement in persons with severe motor impairment: a series of case reports. AB - The purpose of this article is to describe the impact of an intervention involving safe-laser pointing technology on six persons with locked-in syndrome. When these individuals were invited to participate in this project (4 weeks to 18 years post onset), none were able to speak and none were able to access an augmentative and alternative communication (AAC) device. All communicated using eye movements (e.g., looking up or down), eye blinks, dependent scanning strategies with eye movement signals, or eye linking. Following intervention with the Safe-Laser Access System, three of the six participants developed head movement sufficient to control AAC technology. Two participants continue to develop head control; however, their progress has been slowed by repeated illnesses. One participant has discontinued his involvement with the project because of medical and psychological concerns. These six participants represent consecutive referrals to the project. PMID- 17114166 TI - AAC menu interface: effectiveness of active versus passive learning to master abbreviation-expansion codes. AB - This study investigated the accuracy with which 30 young adults without disabilities learned abbreviation expansion codes associated with specific vocabulary items that were stored in an AAC device with two accessing methods: mouse access and keyboard access. Both accessing methods utilized a specialized computer application, called AAC Menu, which allowed for errorless practice. Mouse access prompted passive learning, whereas keyboard access prompted active learning. Results revealed that participants who accessed words via a keyboard demonstrated significantly higher mastery of abbreviation-expansion codes than those who accessed words via a computer mouse. PMID- 17114167 TI - Perspectives of speech language pathologists regarding success versus abandonment of AAC. AB - This three-phase investigation used focus groups and a survey to identify factors that perceived by speech language pathologists as being related to long-term success versus inappropriate abandonment of augmentative and alternative communication (AAC) systems. Factors deemed most important by six focus groups were included in a 106-question survey that was returned by 275 ASHA Special Interest Division #12 (AAC) members. Factor analysis indicated the constructs of Support, Attitude, and System characteristics and Fit as most important to the long-term success of AAC systems. The constructs of Not Maintaining/Adjusting the System, Attitude, Lack of Training, Lack of Support, and Poor Fit were most often related to inappropriate abandonment of AAC systems. Systematic implementation of intervention targeting the constructs is recommended. PMID- 17114168 TI - Comparison of standardized assessments for cognitive and receptive communication skills in young children with complex communication needs. AB - This study compared estimates of receptive language development for children with complex communication needs to estimates of related skill domains. Subscores of the Battelle Developmental Inventory were compared with each other and the MacArthur Communication Developmental Inventory for 41 children with severe expressive impairments over a period of 18 months, beginning at ages 12-24 months. Across all ages studied, children with severe expressive impairments had better receptive language scores than expected for either their cognitive or overall developmental age equivalence scores. While receptive language measures were correlated with each other and age, scores for receptive language skills on the MacArthur were consistently more stringent than for the Battelle for this population. Implications for clinical application of these measures for children with expressive impairments are discussed. PMID- 17114169 TI - Effects of repeated listening experiences on the perception of synthetic speech by individuals with mild-to-moderate intellectual disabilities. AB - The purpose of this study was to examine whether individuals with intellectual disabilities (n = 18) demonstrate improvement in the perception of synthetic speech as a result of repeated exposure to it. Specifically, effects of training on novel versus repeated stimuli produced by the ETI Eloquence speech synthesizer were analyzed. Results revealed that training and experimental task each played a significant (p values < 0.01) role in the perception of synthetic words and sentences. Further, there was an absence of significant effect (p > 0.01) for stimulus type (i.e., repeated versus novel) suggesting that individuals with intellectual disabilities are able to generalize their knowledge of the acoustic phonetic properties of synthetic speech to novel stimuli. Data are also presented for typical participants to establish a benchmark condition for the newly developed Eloquence synthesizer. PMID- 17114170 TI - Preschoolers' speed of locating a target symbol under different color conditions. AB - A pressing decision in AAC concerns the organization of aided visual symbols. One recent proposal suggested that basic principles of visual processing may be important determinants of how easily a symbol is found in an array, and that this, in turn will influence more functional outcomes like symbol identification or use. This study examined the role of color on accuracy and speed of symbol location by 16 preschool children without disabilities. Participants searched for a target stimulus in an array of eight stimuli. In the same-color condition, the eight stimuli were all red; in the guided search condition, four of the stimuli were red and four were yellow; in the unique-color condition, all stimuli were unique colors. Accuracy was higher and reaction time was faster when stimuli were unique colors than when they were all one color. Reaction time and accuracy did not differ under the guided search and the color-unique conditions. The implications for AAC are discussed. PMID- 17114171 TI - Post-school quality of life for individuals with developmental disabilities who use AAC. AB - Even when augmentative and alternative communication (AAC) interventions enhance the communication skills and educational achievements of students with complex communication needs while they are in school, there is no guarantee that these gains will be maintained following students' transition to adult life. Unfortunately, information on the post-school quality of life and related outcomes of individuals with complex communication needs is scarce. This study addressed this issue by examining the post-school outcomes of eight Canadian individuals with developmental disabilities who used AAC technology while they were in school. Two surveys were used to compile the data: the Quality of Life Profile: People with Physical and Sensory Disabilities (Renwick, Rudman, Raphael, & Brown, 1998) and a Communication Survey designed specifically for this study. Four of the participants and the people who knew them best also participated in brief interviews in which they discussed the positive and negative aspects of their school and post-school experiences. Results indicated that participant outcomes in important life domains were generally discouraging. A high positive correlation was found between quality of life and quality of communication scores, and participants who achieved relatively better outcomes showed evidence of higher communicative competence. However, the majority of participants and their supporters were very dissatisfied with the lack of AAC and other services that were available to them as young adults. The results are discussed in relation to outcomes for adults with development disabilities who use AAC and their implications for future research, practice, and advocacy efforts related to transition planning. PMID- 17114172 TI - Distinct Fermi-momentum-dependent energy gaps in deeply underdoped Bi2212. AB - We used angle-resolved photoemission spectroscopy applied to deeply underdoped cuprate superconductors Bi2Sr2Ca(1-x)YxCu2O8 (Bi2212) to reveal the presence of two distinct energy gaps exhibiting different doping dependence. One gap, associated with the antinodal region where no coherent peak is observed, increased with underdoping, a behavior known for more than a decade and considered as the general gap behavior in the underdoped regime. The other gap, associated with the near-nodal regime where a coherent peak in the spectrum can be observed, did not increase with less doping, a behavior not previously observed in the single particle spectra. We propose a two-gap scenario in momentum space that is consistent with other experiments and may contain important information on the mechanism of high-transition temperature superconductivity. PMID- 17114173 TI - Proteus mirabilis clinical isolate harbouring a new variant of Salmonella genomic island 1 containing the multiple antibiotic resistance region. AB - OBJECTIVES: A clinical isolate of Proteus mirabilis strain 18306, which displayed the multidrug resistance phenotype of Salmonella genomic island 1 (SGI1), was examined for the presence of this island including its multiple antibiotic resistance genomic region. METHODS: P. mirabilis 18306 was isolated in March 2006 from a patient in Palestine with diabetic foot infection. Antibiotic susceptibility tests and various molecular techniques, including PCR, cloning and DNA sequencing were used for detection and characterization of SGI1 in P. mirabilis 18306. RESULTS: P. mirabilis 18306 showed the typical multidrug resistance phenotype of SGI1 as it was resistant to ampicillin, chloramphenicol, streptomycin, sulphonamides and tetracycline, in addition to trimethoprim and nalidixic acid. Molecular characterization showed that P. mirabilis 18306 harboured a structure similar to SGI1, except that the aadA2 gene, which confers resistance to streptomycin and spectinomycin, of standard SGI1 had been replaced with dfrA15, which confers resistance to trimethoprim. Furthermore, the nucleotide sequence of the extrachromosomal circular form of SGI1 in P. mirabilis was found to be identical to that of Salmonella Typhimurium DT104. However, PCR results showed that P. mirabilis 18306 was negative for the left and right junctions which represent the integration sites of SGI1 into Salmonella enterica chromosome. Hence, this new variant of SGI1 may be integrated at a different site into the chromosome of P. mirabilis 18306. Tn1826-derived class 2 integron, which carries only two gene cassettes, sat2 and aadA1, was also identified in this strain. CONCLUSIONS: In this study, we identified a new variant SGI1 containing the multiple resistance genomic region in a multidrug-resistant strain of P. mirabilis. This is the first report for SGI1 in a genus other than Salmonella. PMID- 17114174 TI - Variant Salmonella genomic island 1 antibiotic resistance gene clusters in Salmonella enterica serovar Derby isolates from humans in Taiwan. PMID- 17114175 TI - Efficacy of nitazoxanide, tizoxanide and tizoxanide/albendazole sulphoxide combination against Taenia crassiceps cysts. AB - OBJECTIVES: Neurocysticercosis is a common parasitic disease in the CNS in humans caused by the metacestode Taenia solium, with high incidence in developing countries. Albendazole is the drug of choice. However, a wide interindividual variability in the response has been reported. In order to evaluate alternative treatment options, the in vitro efficacy of nitazoxanide, its main metabolite tizoxanide as well as the tizoxanide and albendazole sulphoxide combination was tested against Taenia crassiceps cysts. METHODS: T. crassiceps cysts were incubated in culture medium containing different concentrations of nitazoxanide, tizoxanide and albendazole sulphoxide (0.037-0.42 microg/mL). The effect of the tizoxanide and albendazole sulphoxide combination was evaluated in a fixed concentration ratio (1:1). Isobolographic analyses were used to define the kind of interaction between drugs. Morphological and ultrastructural alterations over the parasite tissue were observed by light and transmission electron microscopy. RESULTS: Nitazoxanide and tizoxanide exhibited cestocidal activity which was time concentration-dependent. The EC(50) values were 0.15, 0.12 and 0.080 microg/mL for nitazoxanide, tizoxanide and albendazole sulphoxide, respectively. No statistical differences between EC(50) values were found, indicating that nitazoxanide and tizoxanide are equally potent as albendazole sulphoxide. The effect of the tizoxanide and albendazole sulphoxide combination was faster than that observed with each drug alone. Isobolographic analysis showed that the effect of the combination was additive. Nitazoxanide and tizoxanide had an effect on the germinal layer, where lipid droplets were found. Nitazoxanide and tizoxanide produced less damage than albendazole sulphoxide on the germinal layer. After the tizoxanide and albendazole sulphoxide combination, a high accumulation of lipid droplets within the germinal layer of the parasite was found. CONCLUSIONS: Our results suggest that nitazoxanide in combination with albendazole could be useful for treatment of cysticercosis infections. Additional in vivo studies are required to confirm this hypothesis. PMID- 17114176 TI - Immunomodulatory effects of fosfomycin in an endotoxin model in human blood. AB - OBJECTIVES: Although a wide range of therapeutic strategies have been developed to improve the outcome of severe sepsis, a convincing reduction in mortality is lacking. Recently, increasing attention has been paid to immunomodulatory effects of antimicrobials. This study set out to explore the immunomodulatory effects of fosfomycin, a broad-spectrum antibiotic frequently used in septic patients, at the protein and molecular levels in vitro. METHODS: Whole blood from 11 healthy volunteers was incubated with 50 pg/mL endotoxin and 100 microg/mL fosfomycin or physiological sodium chloride for 4 h. Real-time RT-PCR was performed for various pro- and anti-inflammatory cytokines. Concentrations of tumour necrosis factor (TNF)-alpha and interleukin (IL)-6 in the supernatant were measured using a commercially available ELISA. RESULTS: Incubation of human leucocytes with endotoxin increased messenger RNA (mRNA) levels of cytokines several thousand fold compared with baseline. The addition of fosfomycin significantly inhibited mRNA levels of pro-inflammatory cytokines such as IL-1-alpha, IL-6 and TNF-alpha after 2 h (P < 0.01), while no significant reduction was observed for the anti inflammatory cytokines IL-4, IL-10 and IL-13 (P = 0.26). At the protein level, the concentrations of IL-6 and TNF-alpha increased approximately 3000- and 600 fold after 4 h of incubation with lipopolysaccharide as compared with baseline, respectively. Addition of fosfomycin significantly reduced cytokine levels by 56% and 73% for IL-6 and TNF-alpha, respectively. CONCLUSIONS: Fosfomycin extensively decreased mRNA levels and release of pro-inflammatory cytokines in human blood. The broad antimicrobial coverage of fosfomycin and its immunosuppressive effects could be clinically useful in patients with sepsis. PMID- 17114177 TI - Prevalence of efflux activity in low-level macrolide-resistant Campylobacter species. PMID- 17114178 TI - Purification and identification of proteins that bind to the hereditary persistence of fetal hemoglobin -198 mutation in the gamma-globin gene promoter. AB - Expression of the gamma-globin gene is silenced in adult humans. However, certain point mutations in the gamma-globin gene promoter are capable of maintaining expression of this gene during adult erythropoiesis, a condition called non deletion hereditary persistence of fetal hemoglobin (HPFH). Among these, the British form of HPFH carrying a T-->C point mutation at position -198 of the Agamma-globin gene promoter results in 4-10% fetal hemoglobin in heterozygotes. In this study, we used nuclear extracts from murine erythroleukemia cells to purify a protein complex that binds the HPFH -198 gamma-globin gene promoter. Members of this protein complex were identified by mass spectrometry and include DNMT1, the transcriptional coactivator p52, the protein SNEV, and RAP74 (the largest subunit of the general transcription factor IIF). Sp1, which was previously considered responsible for HPFH -198 gamma-globin gene activation, was not identified. The potential role of these proteins in the reactivation and/or maintenance of gamma-globin gene expression in the adult transcriptional environment is discussed. PMID- 17114179 TI - Gamma-tocotrienol inhibits nuclear factor-kappaB signaling pathway through inhibition of receptor-interacting protein and TAK1 leading to suppression of antiapoptotic gene products and potentiation of apoptosis. AB - Unlike the tocopherols, the tocotrienols, also members of the vitamin E family, have an unsaturated isoprenoid side chain. In contrast to extensive studies on tocopherol, very little is known about tocotrienol. Because the nuclear factor kappaB (NF-kappaB) pathway has a central role in tumorigenesis, we investigated the effect of gamma-tocotrienol on the NF-kappaB pathway. Although gamma tocotrienol completely abolished tumor necrosis factor alpha (TNF)-induced NF kappaB activation, a similar dose of gamma-tocopherol had no effect. Besides TNF, gamma-tocotrienol also abolished NF-kappaB activation induced by phorbol myristate acetate, okadaic acid, lipopolysaccharide, cigarette smoke, interleukin 1beta, and epidermal growth factor. Constitutive NF-kappaB activation expressed by certain tumor cells was also abrogated by gamma-tocotrienol. Reducing agent had no effect on the gamma-tocotrienol-induced down-regulation of NF-kappaB. Mevalonate reversed the NF-kappaB inhibitory effect of gamma-tocotrienol, indicating the role of hydroxymethylglutaryl-CoA reductase. Gamma-tocotrienol blocked TNF-induced phosphorylation and degradation of IkappaBalpha through the inhibition of IkappaBalpha kinase activation, thus leading to the suppression of the phosphorylation and nuclear translocation of p65. gamma-Tocotrienol also suppressed NF-kappaB-dependent reporter gene transcription induced by TNF, TNFR1, TRADD, TRAF2, TAK1, receptor-interacting protein, NIK, and IkappaBalpha kinase but not that activated by p65. Additionally, the expressions of NF-kappaB regulated gene products associated with antiapoptosis (IAP1, IAP2, Bcl-xL, Bcl-2, cFLIP, XIAP, Bfl-1/A1, TRAF1, and Survivin), proliferation (cyclin D1, COX2, and c-Myc), invasion (MMP-9 and ICAM-1), and angiogenesis (vascular endothelial growth factor) were down-regulated by gamma-tocotrienol. This correlated with potentiation of apoptosis induced by TNF, paclitaxel, and doxorubicin. Overall, our results demonstrate that gamma-tocotrienol inhibited the NF-kappaB activation pathway, leading to down-regulation of various gene products and potentiation of apoptosis. PMID- 17114180 TI - The molecular basis for ligand specificity in a mouse olfactory receptor: a network of functionally important residues. AB - Sequence differences between members of the mouse olfac-tory receptor MOR42 subfamily (MOR42-3 and MOR42-1) are likely to be the basis for variation in ligand binding preference among these receptors. We investigated the specificity of MOR42-3 for a variety of dicarboxylic acids. We used site-directed mutagenesis, guided by homology modeling and ligand docking studies, to locate functionally important residues. Receptors were expressed in Xenopus oocytes and assayed using high throughput electrophysiology. The importance of the Val-113 residue, located deep within the receptor, was analyzed in the context of interhelical interactions. We also screened additional residues predicted to be involved in ligand binding site, based on comparison of ortholog/paralog pairs from the mouse and human olfactory receptor genomes (Man, O., Gilad, Y., and Lancet, D. (2004) Protein Sci. 13, 240-254). A network of 8 residues in transmembrane domains III, V, and VI was identified. These residues form part of the ligand binding pocket of MOR42-3. C12 dicarboxylic acid did not activate the receptor in our functional assay, yet our docking simulations predicted its binding site in MOR42-3. Binding without activation implied that C12 dicarboxylic acid might act as an antagonist. In our functional assay, C12 dicarboxylic acid did indeed act as an antagonist of MOR42-3, in agreement with molecular docking studies. Our results demonstrate a powerful approach based on the synergy between computational predictions and physiological assays. PMID- 17114181 TI - Regulatory effects of mammalian target of rapamycin-activated pathways in type I and II interferon signaling. AB - The mechanisms regulating initiation of mRNA translation for the generation of protein products that mediate interferon (IFN) responses are largely unknown. We have previously shown that both Type I and II IFNs engage the mammalian target of rapamycin (mTOR), resulting in downstream phosphorylation and deactivation of the translational repressor 4E-BP1 (eIF4E-binding protein 1). In the current study, we provide direct evidence that such regulation of 4E-BP1 by IFNalpha or IFNgamma results in sequential dissociation of 4E-BP1 from eukaryotic initiation factor-4E and subsequent formation of a functional complex between eukaryotic initiation factor-4E and eukaryotic initiation factor-4G, to allow initiation of mRNA translation. We also demonstrate that the induction of key IFNalpha- or IFNgamma inducible proteins (ISG15 (interferon-stimulated gene 15) and CXCL10) that mediate IFN responses are enhanced in 4E-BP1 (4E-BP1(-/-)) knockout MEFs, as compared with wild-type 4E-BP1(+/+) MEFs. On the other hand, IFN-dependent transcriptional regulation of the Isg15 and Cxcl10 genes is intact in the absence of 4E-BP1, as determined by real time reverse transcriptase-PCR assays and promoter assays for ISRE and GAS, establishing that 4E-BP1 plays a selective negative regulatory role in IFN-induced mRNA translation. Interestingly, the induction of expression of ISG15 and CXCL10 proteins by IFNs was also strongly enhanced in cells lacking expression of the tuberin (TSC2(-/-)) or hamartin (TSC1(-/-)) genes, consistent with the known negative regulatory effect of the TSC1-TSC2 complex on mTOR activation. In other work, we demonstrate that the induction of an IFN-dependent antiviral response is strongly enhanced in cells lacking expression of 4E-BP1 and TSC2, demonstrating that these elements of the IFN-activated mTOR pathway exhibit important regulatory effects in the generation of IFN responses. Taken altogether, our data suggest an important role for mTOR dependent pathways in IFN signaling and identify 4E-BP1 and TSC1-TSC2 as key components in the generation of IFN-dependent biological responses. PMID- 17114182 TI - Knock-out of the plastid-encoded PetL subunit results in reduced stability and accelerated leaf age-dependent loss of the cytochrome b6f complex. AB - The cytochrome-b6f complex, a key component of the photosynthetic electron transport chain, contains a number of very small protein subunits whose functions are not well defined. Here we have investigated the function of the 31-amino acid PetL subunit encoded in the chloroplast genome in all higher plants. Chloroplast transformed petL knock-out tobacco plants display no obvious phenotype, suggesting that PetL is not essential for cytochrome b6f complex biogenesis and function (Fiebig, A., Stegemann, S., and Bock, R. (2004) Nucleic Acids Res. 32, 3615-3622). We show here that, whereas young mutant leaves accumulate comparable amounts of cytochrome b6f complex and have an identical assimilation capacity as wild type leaves, both cytochrome b6f complex contents and assimilation capacities of mature and old leaves are strongly reduced in the mutant, indicating that the cytochrome b6f complex is less stable than in the wild type. Reduced complex stability was also confirmed by in vitro treatments of isolated thylakoids with chaotropic reagents. Adaptive responses observed in the knockout mutants, such as delayed down-regulation of plastocyanin contents, indicate that plants can sense the restricted electron flux to photosystem I yet cannot compensate the reduced stability of the cytochrome b6f complex by adaptive up regulation of complex synthesis. We propose that efficient cytochrome b6f complex biogenesis occurs only in young leaves and that the capacity for de novo synthesis of the complex is very low in mature and aging leaves. Gene expression analysis indicates that the ontogenetic down-regulation of cytochrome b6f complex biogenesis occurs at the post-transcriptional level. PMID- 17114183 TI - Ligand dynamics in an electron transfer protein. Picosecond geminate recombination of carbon monoxide to heme in mutant forms of cytochrome c. AB - Substitution of the heme coordination residue Met-80 of the electron transport protein yeast iso-1-cytochrome c allows external ligands like CO to bind and thus increase the effective redox potential. This mutation, in principle, turns the protein into a quasi-native photoactivable electron donor. We have studied the kinetic and spectral characteristics of geminate recombination of heme and CO in a series of single M80X (X = Ala, Ser, Asp, Arg) mutants, using femtosecond transient absorption spectroscopy. In these proteins, all geminate recombination occurs on the picosecond and early nanosecond time scale, in a multiphasic manner, in which heme relaxation takes place on the same time scale. The extent of geminate recombination varies from >99% (Ala, Ser) to approximately 70% (Arg), the latter value being in principle low enough for electron injection studies. The rates and extent of the CO geminate recombination phases are much higher than in functional ligand-binding proteins like myoglobin, presumably reflecting the rigid and hydrophobic properties of the heme environment, which are optimized for electron transfer. Thus, the dynamics of CO recombination in cytochrome c are a tool for studying the heme pocket, in a similar way as NO in myoglobin. We discuss the differences in the CO kinetics between the mutants in terms of the properties of the heme environment and strategies to enhance the CO escape yield. Experiments on double mutants in which Phe-82 is replaced by Asp or Gly as well as the M80D substitution indicate that such steric changes substantially increase the motional freedom-dissociated CO. PMID- 17114184 TI - The Hemolymph of the ascidian Styela plicata (Chordata-Tunicata) contains heparin inside basophil-like cells and a unique sulfated galactoglucan in the plasma. AB - The hemolymph of ascidians (Chordata-Tunicata) contains different types of hemocytes embedded in a liquid plasma. In the present study, heparin and a sulfated heteropolysaccharide were purified from the hemolymph of the ascidian Styela plicata. The heteropolysaccharide occurs free in the plasma, is composed of glucose ( approximately 60%) and galactose ( approximately 40%), and is highly sulfated. Heparin, on the other hand, occurs in the hemocytes, and high performance liquid chromatography of the products formed by degradation with specific lyases revealed that it is composed mainly by the disaccharides DeltaUA(2SO(4))-1-->4-beta-d-GlcN(SO(4)) (39.7%) and DeltaUA(2SO(4))-1-->4-beta-d GlcN(SO(4))(6SO(4)) (38.2%). Small amounts of the 3-O-sulfated disaccharides DeltaUA(2SO(4))-1-->4-beta-d-GlcN(SO(4))(3SO(4)) (9.8%) and DeltaUA(2SO(4))-1-->4 beta-d-GlcN(SO(4))(3SO(4))(6SO(4)) (3.8%) were also detected. These 3-O-sulfated disaccharides were demonstrated to be essential for the binding of the hemocyte heparin to antithrombin III. Electron microscopy techniques were used to characterize the ultrastructure of the hemocytes and to localize heparin and histamine in these cells. At least five cell types were recognized and classified as univacuolated and multivacuolated cells, amebocytes, hemoblasts, and granulocytes. Immunocytochemistry showed that heparin and histamine co-localize in intracellular granules of only one type of hemocyte, the granulocyte. These results show for the first time that in ascidians, a sulfated galactoglucan circulates free in the plasma, and heparin occurs as an intracellular product of a circulating basophil-like cell. PMID- 17114185 TI - Structural characterization of human 8-oxoguanine DNA glycosylase variants bearing active site mutations. AB - The human 8-oxoguanine DNA glycosylase (hOGG1) protein is responsible for initiating base excision DNA repair of the endogenous mutagen 8-oxoguanine. Like nearly all DNA glycosylases, hOGG1 extrudes its substrate from the DNA helix and inserts it into an extrahelical enzyme active site pocket lined with residues that participate in lesion recognition and catalysis. Structural analysis has been performed on mutant versions of hOGG1 having changes in catalytic residues but not on variants having altered 7,8-dihydro-8-oxoguanine (oxoG) contact residues. Here we report high resolution structural analysis of such recognition variants. We found that Ala substitution at residues that contact the phosphate 5' to the lesion (H270A mutation) and its Watson-Crick face (Q315A mutation) simply removed key functionality from the contact interface but otherwise had no effect on structure. Ala substitution at the only residue making an oxoG-specific contact (G42A mutation) introduced torsional stress into the DNA contact surface of hOGG1, but this was overcome by local interactions within the folded protein, indicating that this oxoG recognition motif is "hardwired." Introduction of a side chain intended to sterically obstruct the active site pocket (Q315F mutation) led to two different structures, one of which (Q315F(*149)) has the oxoG lesion in an exosite flanking the active site and the other of which (Q315F(*292)) has the oxoG inserted nearly completely into the lesion recognition pocket. The latter structure offers a view of the latest stage in the base extrusion pathway yet observed, and its lack of catalytic activity demonstrates that the transition state for displacement of the lesion base is geometrically demanding. PMID- 17114186 TI - ApoA-I mimetic peptides with differing ability to inhibit atherosclerosis also exhibit differences in their interactions with membrane bilayers. AB - Two homologous apoA-I mimetic peptides, 3F-2 and 3F(14), differ in their in vitro antiatherogenic properties (Epand, R. M., Epand, R. F., Sayer, B. G., Datta, G., Chaddha, M., and Anantharamaiah, G. M. (2004) J. Biol. Chem. 279, 51404-51414). In the present work, we demonstrate that the peptide 3F-2, which has more potent anti-inflammatory activity in vitro when administered intraperitoneally to female apoE null mice (20 microg/mouse/day) for 6 weeks, inhibits atherosclerosis (lesion area 15,800 +/- 1000 microm(2), n = 29), whereas 3F(14) does not (lesion area 20,400 +/- 1000 microm(2), n = 26) compared with control saline administered (19,900 +/- 1400 microm(2), n = 22). Plasma distribution of the peptides differs in that 3F-2 preferentially associates with high density lipoprotein, whereas 3F(14) preferentially associates with apoB-containing particles. After intraperitoneal injection of (14)C-labeled peptides, 3F(14) reaches a higher maximal concentration and has a longer half-time of elimination than 3F-2. A study of the effect of these peptides on the motional and organizational properties of phospholipid bilayers, using several NMR methods, demonstrates that the two peptides insert to different extents into membranes. 3F-2 with aromatic residues at the center of the nonpolar face partitions closer to the phospholipid head group compared with 3F(14). In contrast, only 3F(14) affects the terminal methyl group of the acyl chain, decreasing the (2)H order parameter and at the same time also decreasing the molecular motion of this methyl group. This dual effect of 3F(14) can be explained in terms of the cross-sectional shape of the amphipathic helix. These results support the proposal that the molecular basis for the difference in the biological activities of the two peptides lies with their different interactions with membranes. PMID- 17114187 TI - Enhanced upper respiratory tract airflow and head fanning reduce brain temperature in brain-injured, mechanically ventilated patients: a randomized, crossover, factorial trial. AB - BACKGROUND: Heat loss from the upper airways and through the skull are physiological mechanisms of brain cooling which have not been fully explored clinically. METHODS: This randomized, crossover, factorial trial in 12 brain injured, orally intubated patients investigated the effect of enhanced nasal airflow (high flow unhumidified air with 20 p.p.m. nitric oxide gas) and bilateral head fanning on frontal lobe brain temperature and selective brain cooling. After a 30 min baseline, each patient received the four possible combinations of the interventions--airflow, fanning, both together, no intervention--in randomized order. Each combination was delivered for 30 min and followed by a 30 min washout, the last 5 min of which provided the baseline for the next intervention. RESULTS: The difference in mean brain temperature over the last 5 min of the preceding washout minus the mean over the last 5 min of intervention, was 0.15 degrees C with nasal airflow (P=0.001, 95% CI 0.06-0.23 degrees C) and 0.26 degrees C with head fanning (P<0.001, 95% CI 0.17-0.34 degrees C). The estimate of the combined effect of airflow and fanning on brain temperature was 0.41 degrees C. Selective brain cooling did not occur. CONCLUSION: Physiologically, this study demonstrates that heat loss through the upper airways and through the skull can reduce parenchymal brain temperature in brain-injured humans and the onset of temperature reduction is rapid. Clinically, in ischaemic stroke, a temperature decrease of 0.27 degrees C may reduce the relative risk of poor outcome by 10-20%. Head fanning may have the potential to achieve a temperature decrease of this order. PMID- 17114188 TI - Infliximab maintains a high degree of clinical response in patients with active psoriatic arthritis through 1 year of treatment: results from the IMPACT 2 trial. AB - OBJECTIVE: To evaluate the efficacy and safety of infliximab through 1 year in patients with psoriatic arthritis (PsA) enrolled in the IMPACT 2 trial. METHODS: In this double blind, placebo controlled, phase III study, 200 patients with active PsA were randomised to receive infusions of infliximab 5 mg/kg or placebo at weeks 0, 2, 6, and every 8 weeks thereafter through 1 year. Patients with persistent disease activity could enter early escape at week 16, and all remaining placebo patients crossed over to infliximab at week 24. Patients randomised to infliximab who had no response or who lost response could escalate their dose to 10 mg/kg starting at week 38. Clinical efficacy was assessed based on the proportion of patients achieving ACR 20 and PASI 75 responses. Major clinical response (that is, maintenance of ACR 70 response for 24 continuous weeks) was assessed for the first time in PsA. RESULTS: Through 1 year of treatment, 58.9% and 61.4% of patients in the randomised infliximab and placebo/infliximab groups, respectively, achieved ACR 20; corresponding figures for PASI 75 were 50.0% and 60.3%. At week 54, major clinical response was achieved by 12.1% of patients in the infliximab group. The safety profile of infliximab through week 54 was consistent with that seen through week 24. Two malignancies occurred: basal cell skin cancer (placebo) and stage I Hodgkin's lymphoma (infliximab). CONCLUSION: Infliximab maintains a high degree of clinical efficacy and continues to be well tolerated in patients with PsA through 1 year of treatment. PMID- 17114189 TI - T cell responses to a non-glycosylated epitope predominate in type II collagen immunised HLA-DRB1*0101 transgenic mice. AB - AIM: To study collagen-induced arthritis in human leucocyte antigen (HLA)-DR1 transgenic mice lacking endogenous major histocompatibility complex class II molecules (MHC-II) and to determine T cell specificity against the arthritogenic CII(259-273) epitope of type II collagen either unmodified or post translationally glycosylated at Lys(264). METHODS: Arthritis was induced by immunisation with human type II collagen in complete Freund's adjuvant and measured by footpad swelling, clinical score and histology. T cell responses were assessed by proliferation of spleen and lymph node cells and in antigen presentation assays, using T cell hybridomas specific for the glycosylated and non-glycosylated CII(259-273) epitope. RESULTS: The incidence of arthritis was 50% in DR1-transgenic mice lacking endogenous MHC-II molecules. Recall T cell responses in draining lymph nodes and spleen were consistently greater against the non-glycosylated epitope than to the glycosylated CII(259-273). Most of the T cell hybridomas generated from CII-immunised mice recognised the non-glycosylated CII epitope and this form of the epitope was also presented with 100-fold higher efficiency and 1 h faster kinetics by both macrophages and dendritic cells. CONCLUSION: This study shows that T cell responses to the non-glycosylated epitope of heterologous (human) CII are dominant in HLA-DR1 transgenic mice lacking MHC-II, which could contribute to the pathogenicity of autoimmune arthritis. PMID- 17114190 TI - A randomised, double-blind, placebo-controlled trial of a recombinant version of human alpha-fetoprotein (MM-093) in patients with active rheumatoid arthritis. AB - BACKGROUND: Rheumatoid arthritis (RA) tends to remit during pregnancy, with more patients achieving remission in the third trimester, coinciding with an increase in levels of alpha-fetoprotein (AFP). In vitro and animal studies have shown that AFP has immunomodulatory properties. MM-093 is a non-glycosylated, recombinant version of human AFP. OBJECTIVE: To assess the safety, tolerability and clinical effects of MM-093 during a 12-week, randomised, double-blind, placebo-controlled study. METHODS: 12 patients with RA, who had active disease and were on stable doses of methotrexate, received weekly subcutaneous injections of placebo or 21 mg of MM-093. Assessments were carried out at baseline and weekly thereafter. RESULTS: Baseline characteristics were similar in both groups. There was one dropout in the placebo group, due to flare of disease. Treatment with MM-093 was well tolerated. No serious adverse event was observed. By day 85, MM-093 produced a significant mean improvement from baseline in Disease Activity Score 28 (DAS28; 0.913 vs 0.008, p = 0.033) and patient's global assessment (28.9% vs -36.3%, p = 0.02) compared with placebo. CONCLUSION: This is the first randomised, controlled trial of MM-093, a recombinant version of human AFP, in patients with RA. MM-093 was well tolerated. Evidence of efficacy was observed, suggesting that MM-093 may have therapeutic potential in RA. PMID- 17114191 TI - Does knee pain in the community behave like a regional pain syndrome? Prospective cohort study of incidence and persistence. AB - OBJECTIVES: To investigate whether knee pain in the community behaves like a regional pain syndrome, determined by its association with mental health, self rated health (SRH) and beliefs about prognosis. METHODS: An 18-month postal follow-up was conducted in 1798 working-aged subjects, sampled from the community. At baseline questions were asked about pain in the knee lasting > or =1 day in the previous 12 months, mental health (Short-Form 36), somatising tendency (elements of the Brief Symptom Inventory), SRH and concern about 12 month prognosis. At follow-up we asked about knee pain during the last 4 weeks, and whether it had been present for > or =14 days or prescription-treated. Associations with incidence and persistence were explored using logistic regression. RESULTS: The 1256 participants (70% response) comprised 468 with knee pain at baseline and 788 without. Among the former, 49% had persistent knee pain at follow-up, while among the latter, 15% reported new symptoms. Incident prescription-treated knee pain was strongly associated with all of the mental health variables and with SRH. The odds of knee pain persisting were significantly raised in the least versus most favourable bands of somatising tendency and SRH, and persistence was also significantly more common among those who at baseline were concerned that they would still have a problem in 12 months. CONCLUSIONS: Our observations support the hypothesis that knee pain in the community shares risk factors in common with other non-specific regional pain syndromes. PMID- 17114192 TI - Reduced telomere length in rheumatoid arthritis is independent of disease activity and duration. AB - BACKGROUND: Rheumatoid arthritis (RA) is associated with reduced lifespan and shortened telomere length in lymphocytes, but the mechanism underlying this is unclear. Telomere loss in white blood cells (WBC) is accelerated by oxidative stress and inflammation in vitro. It was postulated that the accelerated WBC telomere shortening in RA occurs as a result of exposure to chronic inflammation. OBJECTIVES: To measure telomere terminal restriction fragment (TRF) length in a large cohort of RA cases and healthy controls, to explore associations of TRF length with features of disease and with RA-associated HLA-DRB1 alleles. METHODS: WBC and TRF length were measured by Southern blot in DNA from 176 hospital-based RA cases satisfying the 1987 American College of Rheumatology criteria and from 1151 controls. TRF length was compared between cases and controls, and the effects of disease duration, severity and HLA-DRB1 alleles encoding the shared epitope (SE) were assessed. RESULTS: Age- and sex-adjusted TRF length was significantly shorter in RA cases compared with controls (p<0.001). There was no association between age- and sex-adjusted TRF length and disease duration, C reactive protein or Larsen score. The presence of one or more SE-encoding alleles was associated with reduced adjusted TRF length in RA cases (SE positive vs SE negative cases, p=0.038), but not in controls. CONCLUSION: The reduced TRF length in a large group of patients with RA compared with controls has been shown. The reduction is apparently independent of disease duration and markers of disease severity, but is influenced by HLA-DRB1 genotype. PMID- 17114193 TI - Effect of different sites for cryopreserved ovarian tissue implantation in rabbit. AB - BACKGROUND: Autotransplantation of frozen-thawed ovarian tissue has proven to be an effective method to restore endocrine function and fertility. But it remains to be studied which site and which method is most effective and practical. We therefore implanted small pieces of cryopreserved ovarian tissues into different sites in rabbits to find the optimal position. METHODS: Fifteen New Zealand white female rabbits were randomly divided into three groups. In group 1, fresh ovarian tissues were implanted into the mesometrium and ovarian bursa. In group 2, cryopreserved ovarian tissues were implanted into the mesometrium and ovarian bursa. In group 3, cryopreserved ovarian tissues were implanted into the preserved ovary. RESULTS: There were no significant differences among the three groups as to the proportions of normal and morphologically changed follicles in implanted ovarian tissues. The implanted ovarian tissues in the three groups did not show any evident changes in histology and ultrastructure, and all resumed follicle development and revealed maturescent follicles. CONCLUSIONS: Cryopreservation and implantation of small pieces of ovarian tissues are feasible. Generally, the mesometrium, ovarian bursa and ovary are all available sites for implantation and have similar rates of acceptance, despite some differences in the details of implantation. PMID- 17114194 TI - Stimulation of embryo hatching and implantation by prostacyclin and peroxisome proliferator-activated receptor delta activation: implication in IVF. AB - BACKGROUND: Successful IVF depends in part on quality embryos. Recent work suggests that prostaglandin I(2) (PGI(2) or prostacyclin) promotes the development of embryos in vitro and enhances their implantation potential. The mechanism underlying the effects of PGI(2) is unclear. It has been reported that peroxisome proliferator-activated receptor delta (PPARdelta) mediates the effects of PGI(2) at the implantation sites. METHODS: The expression of PPARdelta in the preimplantation embryos was examined by RT-PCR, western blot analysis and immunohistochemistry. Synthetic PPARdelta ligand (L-165041) and PPARdelta targeted (PPARdelta(-/-)) embryos were used to reveal the roles of PPARdelta in PGI(2)-stimulated and spontaneous embryo development. RESULTS: Preimplantation embryos express PPARdelta, which is essential for the enhancing effect of PGI(2) and the spontaneous progression of preimplantation embryos. Enhanced blastocyst hatching by PGI(2) (P < 0.05) was abrogated by PPARdelta deletion. Blastocyst formation and embryo hatching were impaired in PPARdelta(-/-) embryos. PPARdelta deletion significantly reduced embryo cell proliferation (P < 0.01); PPARdelta activation increased embryo cell proliferation (P < 0.05). PPARdelta activation enhanced the implantation of wild-type (WT) embryos (P < 0.05); PPARdelta deletion reduced embryo implantation (P < 0.05). CONCLUSIONS: PPARdelta is essential for spontaneous and PGI(2)-stimulated embryo development and blastocyst hatching. The implantation of cultured embryos is enhanced by PPARdelta activation. PPARdelta represents a novel therapeutic target to improve IVF outcome. PMID- 17114195 TI - Novel disulphide esters of carbothioic acid as potent, non-detergent spermicides with low toxicity to Lactobacillus and HeLa cells in vitro. AB - BACKGROUND: The design, synthesis, characterization and evaluation of a novel series of non-detergent spermicides has led to the discovery of two unique molecules (DSE-36 and DSE-37) that were approximately 25 times more potent spermicides than nonoxynol-9 (N-9). METHODS: Normal human spermatozoa were used to assess the spermicidal activity (Sander-Cramer Assay), the effect on sperm membrane integrity [hypo-osmotic swelling test (HOST)], supravital staining and scanning electron microscopy (SEM) and the induction of apoptosis [fluorescein isothiocyanate (FITC) Annexin-V and JC-1 labelling using flow cytometry] by the new class of compounds. HeLa and Lactobacillus cultures were used to assess the cytotoxicity of compounds and their compatibility to normal vaginal flora, respectively. RESULTS: Compounds DSE-36 and DSE-37 exhibited a strong spermicidal activity [minimum effective concentration (MEC) = 0.002%], which was approximately 25 times more potent than that of N-9 and Sapindus saponins (MEC = 0.05%). As compared with surfactants, DSE-36 and DSE-37 were found to be safer at MEC towards the growth and survival of Lactobacilli and HeLa cells in vitro and to have a milder effect on sperm plasma membrane. At EC(50) both induced apoptosis in sperm cells as characterized by increased labelling with Annexin-V and decreased polarization of sperm mitochondria. CONCLUSION: Preliminary studies have revealed that in sharp contrast to the non-specific surfactant action of N 9, DSE-36 and DSE-37 have a highly potent, mechanism-based, detrimental action on human sperm. The unique ability of these non-detergent molecules to selectively kill sperm and spare Lactobacilli and HeLa cells at MEC values much lower than that required for N-9 indicates their potential as superior ingredients for formulation into microbicidal contraceptives. PMID- 17114196 TI - Influence of follicular fluid meiosis-activating sterol on aneuploidy rate and precocious chromatid segregation in aged mouse oocytes. AB - BACKGROUND: Follicular fluid meiosis-activating sterol (FF-MAS) protects young oocytes from precocious chromatid separation (predivision). Reduced expression of cohesion and checkpoint proteins and predivision has been hypothesized to occur in age-related aneuploidy in oocytes. METHODS: To know whether FF-MAS also protects aged oocytes from predivision and from age-related non-disjunction, we analysed chromosome constitution in mouse oocytes matured spontaneously with or without 10 microM FF-MAS and in hypoxanthine (HX)-arrested young and aged oocytes induced to resume maturation by FF-MAS. Messenger RNA for checkpoint protein MAD2 and cohesion protein SMC1beta was compared between oocytes matured with or without FF-MAS. RESULTS: Aged oocytes possessed many bivalents with single distal chiasma at meiosis I. Predivision was especially high in aged oocytes cultured sub-optimally to metaphase II in alpha-minimum essential medium (alpha-MEM). FF MAS reduced predivision significantly (P < 0.001) but neither reduced non disjunction nor induced aneuploidy in aged oocytes. Polyploidy was high in FF-MAS stimulated maturation, in particular in the aged oocytes (P > 0.001). Relative levels of Smc1beta mRNA appeared increased by maturation in FF-MAS, and mitochondrial clustering was restored. CONCLUSIONS: Sister chromatids of aged oocytes appear to be highly susceptible to precocious chromatid separation, especially when maturation is under sub-optimal conditions, e.g. in the absence of cumulus and FF-MAS. This may relate to some loss of chromatid cohesion during ageing. FF-MAS protects aged oocytes from predivision during maturation, possibly by supporting Smc1beta expression, thus reducing risks of meiotic errors, but it cannot prevent age-related non-disjunction. Aged oocytes appear prone to loss of co-ordination between nuclear maturation and cytokinesis suggesting age-related relaxed cell cycle control. PMID- 17114197 TI - Evaluation of the utility of multiple endocrine and ultrasound measures of ovarian reserve in the prediction of cycle cancellation in a high-risk IVF population. AB - BACKGROUND: Unexpectedly poor response leading to IVF cycle cancellation is a distressing treatment outcome. We have prospectively assessed several markers of ovarian reserve in a high risk IVF population to determine their utility in predicting IVF cycle cancellation. METHODS: Eighty-four women at high risk of cycle cancellation due to raised FSH, previous poor response and/or age > or =40 years attending for high-dose short protocol IVF treatment had baseline measures of FSH, inhibin B, anti-Mullerian hormone (AMH), antral follicle count (AFC) and ovarian volume. A GnRH agonist was then administered and, 24 h later, estradiol (E(2)) and inhibin B measures were repeated. RESULTS: Fifty-seven per cent of patients in this study had a poor response to stimulation, and 15% were cancelled. Using multivariate logistic regression, we found that day 3 inhibin B levels were the best predictor of cycle cancellation with an area under the receiver operating curve (ROC AUC) = 0.78 (P = 0.017). When only considering baseline variables, mean ovarian volume was the best predictor of cycle cancellation (ROC AUC = 0.78; P = 0.016). AMH concentrations were the best predictor of a poor response (P = 0.003), and AMH was also predictive of cycle cancellation (P = 0.007) with very little inter-cycle variability. None of the parameters studied were predictive of ongoing pregnancy. CONCLUSIONS: This group of at-risk patients had a high rate of poor response to simulation and cancellation. Although several measures of ovarian reserve were able to predict cycle cancellation, none were able to predict pregnancy. AMH was predictive of both cycle cancellation and poor response with little inter-cycle variability. PMID- 17114198 TI - C-reactive protein levels predict the incidence of delirium and recovery from it. PMID- 17114199 TI - Endovascular intervention for symptomatic bilateral carotid artery stenosis in an octogenarian. AB - An 89-year-old man presented with two separate minor stroke episodes due to high grade bilateral carotid stenoses, which were successfully treated with endovascular angioplasty and stenting. The role of operative interventions for high grade symptomatic carotid stenosis in patients aged over 80 years is discussed. PMID- 17114200 TI - Is augmentation index a good measure of vascular stiffness in the elderly? AB - OBJECTIVES: we investigated the exact relationship between age and gender on augmentation pressure (AG) and augmentation index (AI) measured over the radial (muscular) and carotid (elastic) arteries. DESIGN AND METHODS: AG is the contribution that wave reflection makes to systolic arterial pressure. AI is an indirect measure of arterial stiffness and is calculated as AG divided by pulse pressure (PP) x100. AG and AI both increase with age. AG and AI were measured in 458 subjects using SphygmoCor. A total of 755 readings were obtained (302 carotid, 453 radial). The mean age was 57.5 +/- 13.7 years. Diabetic subjects were excluded. Among the subjects, 13.5% were hypertensive. RESULTS: statistically, women had mean values of AI significantly higher than men in both radial and carotid arteries. These differences were less marked with AG. Quadratic equations better described the relationship between AI and age but not AG and age. Thus, AI increased with age up to our median age of 55 years but plateaued thereafter, whereas the AG continued to increase steadily with age. A multiple regression analysis demonstrated that both AI and AG were negatively related to height and positively related to diastolic blood pressure (DBP). CONCLUSIONS: AG continues to increase in the elderly over the age of 55, but not AI. AI is higher in women and higher when measured over the carotid than the radial. AI is positively related to DBP and negatively to height. AG is proposed as a more suitable measure of arterial stiffness than AI. PMID- 17114202 TI - The effect of delirium education on use of target PRN medications in older orthopaedic patients. PMID- 17114201 TI - Low serum carotenoids and development of severe walking disability among older women living in the community: the women's health and aging study I. AB - OBJECTIVE: to determine whether low serum carotenoid levels, an indicator of low intake of fruits and vegetables, are associated with the progression of disability in older women. DESIGN: longitudinal analysis in a population-based cohort. SETTING: moderately-severely disabled women, >or=65 years, living in the community in Baltimore, Maryland (the Women's Health and Aging Study I). PARTICIPANTS: 554 women without severe walking disability (inability to walk or walking speed <0.4 m/s) at baseline. MAIN OUTCOME MEASURE: incidence of severe walking disability assessed every 6 months over 3 years. RESULTS: 155 women (27.9%) developed severe walking disability during follow-up. Rates of development of severe walking disability per 100 person-years among women in the lowest and in the three upper quartiles of total carotenoids were, respectively, 13.8 versus 10.9 (P=0.0017). Adjusting for confounders, women in the lowest quartile of total carotenoids were more likely to develop severe walking disability (hazards ratio 1.57, 95% confidence interval 1.24-2.00, P=0.0002) compared with women in the three upper quartiles. CONCLUSION: low serum carotenoid levels, an indicator of low intake of fruits and vegetables, are independent predictors of the progression towards severe walking disability among older women living in the community. PMID- 17114203 TI - Minimal-preparation CT colon in detection of colonic cancer, the Oxford experience. AB - BACKGROUND: the main colonic imaging modalities, including barium enema, colonoscopy and computed tomography colonography, require bowel preparation. Performing these imaging procedures in the elderly can difficult due to immobility, incontinence and poor tolerance of bowel cleansing. Minimal preparation CT (MPCT) colon was introduced in the early 1990s in the UK. Much of the published literature on MPCT colon is limited by small patient numbers and short duration of follow-up. OBJECTIVE: the aim of this study is to review our experience with the MPCT technique involving a large consecutive cohort of patients with long follow-up. METHODS: all studies of MPCT performed in a 1-year period between July 2000 and July 2001 at our institution were reviewed retrospectively. MPCT reports were cross-referenced with the cancer registry to allow for an average period of 30 months follow-up. A definite diagnosis of cancer was only given following the appearance on the cancer registry. Those patients who had negative MPCT colon were assumed to be true negatives if no corresponding name was identified on the cancer registry. In the event of data mismatch, patient notes were reviewed to ascertain a diagnosis. RESULTS: 391 MPCT examinations were performed during the period of the study (209 males, median age 82: age range 56-91 years). Thirty-four patients who had MPCT colon during the study period appeared on the cancer registry. A further three patients with disseminated colorectal malignancy identified on MPCT colon died without histological confirmation (tumour prevalence = 9.5%). Thirty-two of the registry confirmed 34 cases were detected on MPCT colon, giving a sensitivity of 0.94 (95% confidence interval 0.86-1.00). Including the three cases without histological confirmation gives a slightly higher sensitivity of 0.95. There were seven patients with definitely abnormal MPCT colons, who did not appear on the registry, resulting in specificity for definite abnormality of 0.98 (confidence interval 0.97-1.0). However, three of these seven are those who died of disseminated colorectal malignancy as above, raising the specificity to 0.99. Fourteen cases (3.5%) of extra-colonic malignancies were observed in this study. CONCLUSION: even with the longer follow-up of this large cohort of patients the sensitivity and specificity in our study for the diagnosis of colorectal cancer with MPCT remains comparable with that of other studies and this technique competes well with other common colonic imaging modalities. PMID- 17114204 TI - Restless arm symptoms as an extension of restless leg syndrome. PMID- 17114205 TI - Intravenous pamidronate treatment of infants with severe osteogenesis imperfecta. AB - OBJECTIVE: Children with the severe forms of osteogenesis imperfecta have in several studies been treated with intravenous pamidronate, but there are only few reports of the effect of early treatment. AIM: To evaluate the effect of treatment started in infancy. METHODS: In a prospective observational study, with a historic control group, intravenous disodium pamidronate (APD) was given as monthly infusions to 11 children with osteogenesis imperfecta aged 3-13 (median 3.6) months, who had severe osteogenesis imperfecta with congenital bowing of the femora and vertebral compression fractures. RESULTS: During treatment of children aged between 3 and 6 (median 4.5) years, dual-energy x ray absorptiometry measurements of the lumbar spine showed a gradual increase in bone density. Bone metabolism parameters in serum (alkaline phosphatase, osteocalcin, procollagen 1 carboxy-terminal peptide, collagen 1 teleopeptide) and in urine (deoxypyridinoline) indicated a decrease in bone turnover. An improvement of mobility was seen and at the latest recording, at the age of 3.3-6.5 (median 4.8) years, the children could all walk. Vertebral remodelling was seen, with increased vertebral height, and no child developed scoliosis, kyphosis or basilar impression. All children required femoral intramedullar rods for fractures, and five needed tibial rodding for extreme curvatures that prevented functional standing and walking. No adverse effects were seen on growth, fracture healing or blood chemistry. CONCLUSIONS: APD is an efficient symptomatic treatment for infants with severe osteogenesis imperfecta, but additional orthopaedic surgery is often needed. Early treatment may prevent scoliosis and basilar impression. Long-term follow-up is important. PMID- 17114206 TI - Association of three isoforms of the meiotic BOULE gene with spermatogenic failure in infertile men. AB - The complex process of spermatogenesis requires the expression and precise coordination of a multitude of genes. Abnormal function of such genes is frequently associated with male infertility. Among these candidates is the human BOULE gene that is a possible fundamental mediator of meiotic transition. In this study, we describe for the first time the existence of three BOULE transcript variants (B1, B2 and B3). We investigated their tissue specificity and mRNA transcript levels in 23 testis biopsies from infertile men. B1, B2 and B3 differed solely in their N-terminal sequences, which are encoded by three alternatively spliced exons 1. In humans, all three isoforms are exclusively expressed in the testes in a relative proportion of 80:220:1 for B1, B2 and B3, respectively. RT-PCR quantification revealed significantly reduced mRNA expression of all three variants in testicular biopsies with meiotic arrest (MA) compared with those with qualitatively complete spermatogenesis. Alteration of the B1/B2 and B1/B3 transcript ratios was correlated with reduced meiotic capacity of spermatocytes to produce round spermatids as assessed by flow cytometry. Furthermore, BOULE mRNA reduction in biopsies with MA paralleled the absence of BOULE protein as analysed by immunohistochemistry. In conclusion, the relative proportions of B1, B2 and B3 may serve as predictive markers for meiotic efficiency and thus the probability of finding haploid cells in the human testis. Among the three isoforms, B2 might have the major role for meiotic completion. PMID- 17114207 TI - Strategies and outcomes of PGD of familial adenomatous polyposis. AB - Owing to adult onset of hereditary cancer, prenatal diagnosis (PND) raises numerous ethical issues on the acceptability to terminate an affected pregnancy (TOP). PND for these disorders is often considered as unacceptable by couples as well as geneticists and legal or ethical authorities, but preimplantation genetic diagnosis (PGD), even if subject to controversy, seems to be a more acceptable option. Therefore, many couples, who do not want to transmit their cancer to their children, consider PGD as their only reproductive option. This article describes our experience of PGD for familial adenomatous polyposis (FAP). Twelve couples were referred between 2000 and 2005. We developed PGD tests to detect the mutation alone, but we rapidly set up multiplex PCR combining mutation detection and indirect diagnosis. Finally, we set up duplex and triplex indirect diagnoses to be able to offer a PGD, whatever mutation was involved in familial cases. PGD strategies were based on (i) a new double allele-specific PCR approach (D-ARMS) allowing the detection of the wild-type and mutated allele; (ii) PCR fragments sizing and (iii) restriction length polymorphisms. For the 12 referrals, we developed eight tests, and 11 cycles have been performed for four couples, resulting in eight embryo transfers and five pregnancies, with the birth of one healthy boy and two ongoing pregnancies. We are now able to propose PGD to most couples at risk of transmitting FAP to their offspring, whether the mutation is familial or occurred de novo. PMID- 17114208 TI - Derivation of oocyte-like cells from a clonal pancreatic stem cell line. AB - Adult pancreatic stem cells (PSCs) are able to differentiate spontaneously in vitro into various somatic cell types. Stem cells isolated from rat pancreas show extensive self-renewal ability and grow in highly viable long-term cultures. Additionally, these cells express typical stem cell markers such as Oct-4, nestin and SSEA-1. Although differentiation potential is slightly decreasing in long term cultures, it is possible to keep cell lines up to passage 140. Clonal cell lines could be established from different passages and showed similar characteristics. Remarkably, one clonal cell line, generated from passage 75, showed deviant properties during further culture. Clonal cells formed aggregates, which built tissue-like structures in suspension culture. These generated 3D aggregates produced permanently new cells at the outside margin. Released cells had remarkable size, and closer examination by light microscopy analysis revealed oocyte-like morphology. A comparison of the gene expression patterns between primary cultures of passages 8 and 75, the clonal cell line and the produced oocyte-like cells (OLCs) from tissue-like structures demonstrated some differences. Expression of various germ cell markers, such as Vasa, growth differentiation marker 9 and SSEA-1, increased in the clonal cell line, and OLCs showed additionally expression of meiosis-specific markers SCP3 and DMC1. We here present a first pilot study investigating the putative germ line potential of adult PSCs. PMID- 17114209 TI - A new paradigm for profiling testicular gene expression during normal and disturbed human spermatogenesis. AB - The aim of this study was to identify gene expression patterns of the testis that correlate with the appearance of distinct stages of male germ cells. We avoided the pitfalls of mixed pathological phenotypes of the testis and circumvented the inapplicability of using the first spermatogenic wave as done previously on rodents. This was accomplished by using 28 samples showing defined and highly homogeneous pathologies selected from 578 testicular biopsies obtained from 289 men with azoospermia (two biopsies each). The molecular signature of the different developmental stages correlated with the morphological preclassification of the testicular biopsies, as shown by resampling-based hierarchical clustering using different measures of variability. By using analysis of variance (ANOVA) and extensive permutation analysis, we filtered 1181 genes that exhibit exceptional statistical significance in testicular expression and grouped subsets with transcriptional changes within the pre-meiotic (348 genes), post-meiotic (81 genes) and terminal differentiation (38 genes) phase. Several distinct molecular classes, metabolic pathways and transcription factor binding sites are involved, depending on the transcriptional profile of the gene clusters that were built using a novel clustering procedure based on not only similarity but also statistical significance. PMID- 17114210 TI - Pandemic obesity in Europe. PMID- 17114211 TI - A model clinical trials agreement. PMID- 17114212 TI - Community based occupational therapy for patients with dementia and their care givers: randomised controlled trial. AB - OBJECTIVE: To determine the effectiveness of community based occupational therapy on daily functioning of patients with dementia and the sense of competence of their care givers. DESIGN: Single blind randomised controlled trial. Assessors were blinded for treatment allocation. SETTING: Memory clinic and day clinic of a geriatrics department and participants' homes. PARTICIPANTS: 135 patients aged > or =65 with mild to moderate dementia living in the community and their primary care givers. INTERVENTIONS: 10 sessions of occupational therapy over five weeks, including cognitive and behavioural interventions, to train patients in the use of aids to compensate for cognitive decline and care givers in coping behaviours and supervision. MAIN OUTCOME MEASURES: Patients' daily functioning assessed with the assessment of motor and process skills (AMPS) and the performance scale of the interview of deterioration in daily activities in dementia (IDDD). Care giver burden assessed with the sense of competence questionnaire (SCQ). Participants were evaluated at baseline, six weeks, and three months. RESULTS: Scores improved significantly relative to baseline in patients and care givers in the intervention group compared with the controls (differences were 1.5 (95% confidence interval 1.3 to 1.7) for the process scale; -11.7 (-13.6 to -9.7) for the performance scale; and (11.0; 9.2 to 12.8) for the competence scale). This improvement was still significant at three months. The number needed to treat to reach a clinically relevant improvement in motor and process skills score was 1.3 (1.2 to 1.4) at six weeks. Effect sizes were 2.5, 2.3, and 1.2, respectively, at six weeks and 2.7, 2.4, and 0.8, respectively, at 12 weeks. CONCLUSIONS: Occupational therapy improved patients' daily functioning and reduced the burden on the care giver, despite the patients' limited learning ability. Effects were still present at 12 weeks, which justifies implementation of this intervention. TRIAL REGISTRATION: Clinical Trials NCT00295152 [ClinicalTrials.gov]. PMID- 17114213 TI - Primary tumor levels of tissue inhibitor of metalloproteinases-1 are predictive of resistance to chemotherapy in patients with metastatic breast cancer. AB - PURPOSE: Only about 50% of metastatic breast cancer patients benefit from cytotoxic chemotherapy. Today, no validated markers exist for prediction of chemotherapy sensitivity/resistance in this patient group. Tissue inhibitor of metalloproteinases-1 (TIMP-1) has been shown to protect against apoptosis, and the purpose of the present study was to test the hypothesis that tumors expressing high levels of TIMP-1 are protected against apoptosis-inducing agents and thus less sensitive to apoptosis-inducing chemotherapeutic drugs. EXPERIMENTAL DESIGN: We investigated the association between primary tumor expression levels of TIMP-1 protein and objective response to first-line chemotherapy in 173 patients with metastatic breast cancer. RESULTS: When analyzed as a continuous log-transformed variable, increasing TIMP-1 levels were significantly associated with lack of response to cyclophosphamide/methotrexate/5 fluorouracil and anthracycline-based chemotherapy (P = 0.01; odds ratio, 2.0; 95% confidence interval, 1.1-3.3). In a multivariate model, including lymph node status, steroid hormone receptor status, menopausal status, dominant metastases site, type of chemotherapy, and disease-free interval, TIMP-1 was significantly associated with resistance to treatment (P = 0.03; odds ratio, 1.7; 95% confidence interval, 1.1-3.3). CONCLUSIONS: In the present exploratory study, we showed that elevated tumor tissue TIMP-1 levels were significantly associated with a poor response to chemotherapy. By using TIMP-1, we identified a group of patients with metastatic breast cancer, which hardly respond to the most frequently used chemotherapy regimes (i.e., cyclophosphamide/methotrexate/5 fluorouracil and anthracyclines). PMID- 17114214 TI - Regulation of cardiac L-type Ca2+ current in Na+-Ca2+ exchanger knockout mice: functional coupling of the Ca2+ channel and the Na+-Ca2+ exchanger. AB - L-type Ca2+ current (I(Ca)) is reduced in myocytes from cardiac-specific Na+-Ca2+ exchanger (NCX) knockout (KO) mice. This is an important adaptation to prevent Ca2+ overload in the absence of NCX. However, Ca2+ channel expression is unchanged, suggesting that regulatory processes reduce I(Ca). We tested the hypothesis that an elevation in local Ca2+ reduces I(Ca) in KO myocytes. In patch clamped myocytes from NCX KO mice, peak I(Ca) was reduced by 50%, and inactivation kinetics were accelerated as compared to wild-type (WT) myocytes. To assess the effects of cytosolic Ca2+ concentration on I(Ca), we used Ba2+ instead of Ca2+ as the charge carrier and simultaneously depleted sarcoplasmic reticular Ca2+ with thapsigargin and ryanodine. Under these conditions, we observed no significant difference in Ba2+ current between WT and KO myocytes. Also, dialysis with the fast Ca2+ chelator BAPTA eliminated differences in both I(Ca) amplitude and decay kinetics between KO and WT myocytes. We conclude that, in NCX KO myocytes, Ca2+-dependent inactivation of I(Ca) reduces I(Ca) amplitude and accelerates current decay kinetics. We hypothesize that the elevated subsarcolemmal Ca2+ that results from the absence of NCX activity inactivates some L-type Ca2+ channels. Modulation of subsarcolemmal Ca2+ by the Na+-Ca2+ exchanger may be an important regulator of excitation-contraction coupling. PMID- 17114215 TI - Domain registration in raft-mimicking lipid mixtures studied using polymer tethered lipid bilayers. AB - The degree of domain registration in a liquid-ordered/liquid-disordered phase separating lipid mixture consisting of 1-stearoyl-2-oleoyl-sn-3-phosphocholine, egg sphingomyelin, and cholesterol (molar mixing ratio of 1:1:1) was studied using three different planar lipid bilayer architectures distinguished by their bilayer-substrate distance d using epifluorescence microscopy. The bilayer systems, which were built layer by layer using Langmuir-Blodgett/Schaefer film depositions, included a solid-supported bilayer (d approximately 15 A) and two polymer-supported bilayers with d approximately 30 A and d approximately 58 A, respectively. Complete domain registration between Langmuir-Blodgett and Schaefer monolayer domains was observed for d approximately 58 A but not in the cases when d approximately 15 A and d approximately 30 A. Building the bilayer layer by layer guaranteed that any preexisting domains were not in registration initially; our data show that the domain registration observed was not caused by lipid flip flop or by lateral rearrangement of preexisting large-scale domains. Instead, additional studies on bilayer systems with asymmetric lipid composition indicate that preexisting domains in the Langmuir-Blodgett monolayer induce the formation of completely registered domains in the opposite Schaefer monolayer. This study provides insight into possible biophysical mechanisms of transbilayer domain coupling. Our findings support the concept that the formation of transbilayer signaling platforms based on registered raft domains may occur without the active involvement of membrane-spanning proteins. PMID- 17114216 TI - Action potential duration dispersion and alternans in simulated heterogeneous cardiac tissue with a structural barrier. AB - Structural barriers to wave propagation in cardiac tissue are associated with a decreased threshold for repolarization alternans both experimentally and clinically. Using computer simulations, we investigated the effects of a structural barrier on the onset of spatially concordant and discordant alternans. We used two-dimensional tissue geometry with heterogeneity in selected potassium conductances to mimic known apex-base gradients. Although we found that the actual onset of alternans was similar with and without the structural barrier, the increase in alternans magnitude with faster pacing was steeper with the barrier--giving the appearance of an earlier alternans onset in its presence. This is consistent with both experimental structural barrier findings and the clinical observation of T-wave alternans occurring at slower pacing rates in patients with structural heart disease. In ionically homogeneous tissue, discordant alternans induced by the presence of the structural barrier arose at intermediate pacing rates due to a source-sink mismatch behind the barrier. In heterogeneous tissue, discordant alternans occurred during fast pacing due to a barrier-induced decoupling of tissue with different restitution properties. Our results demonstrate a causal relationship between the presence of a structural barrier and increased alternans magnitude and action potential duration dispersion, which may contribute to why patients with structural heart disease are at higher risk for ventricular tachyarrhythmias. PMID- 17114217 TI - Disulfide trapping the mechanosensitive channel MscL into a gating-transition state. AB - The mechanosensitive channel of large conductance, MscL, serves as a biological emergency release valve protecting bacteria from acute osmotic downshock, and is to date the best characterized mechanosensitive channel. The N-terminal region of the protein has been shown to be critical for function by random, site-directed, and deletion mutagenesis, yet is structurally poorly understood. One model proposes that the extreme N-termini form a cluster of amphipathic helices that serves as a cytoplasmic second gate, separated from the pore-forming transmembrane domain by a "linker". Here, we have utilized cysteine trapping of single-cysteine mutated channels to determine the proximity, within the homopentameric complex, of residues within and just peripheral to this proposed linker. Our results indicate that all residues in this region can form disulfide bridges, and that the percentage of dimers increases when the channel is gated in vivo. Functional studies suggest that oxidation traps one of these mutated channels, N15C, into a gating-transition state that retains the capacity to obtain both fully open and closed states. The data are not easily explained by current models for the smooth transition from closed-to-open states, but predict that an asymmetric movement of one or more of the subunits commonly occurs upon gating. PMID- 17114218 TI - Differential regulation of ionotropic glutamate receptors. AB - Ionotropic glutamate receptors (iGluRs), a family of ligand-gated ion channels, are responsible for the majority of fast excitatory neurotransmission in the central nervous system. Within this family, different members serve distinct roles at glutamatergic synapses. Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors mediate fast depolarization while N-methyl-D-aspartate (NMDA) receptors mediate the slower component of the excitatory postsynaptic potential. These disparate functions suggest alternate modes of regulation. In this work, we show that endogenous regulators of iGluRs have different abilities to bind to specific domains of NMDA NR1-1b and AMPA GluR2 subunits. We have previously shown that the sulfated neurosteroids pregnenolone sulfate and 3alpha hydroxy-5beta-pregnan-20-one sulfate bind to the extracellular glutamate-binding core (S1S2) of the GluR2 subunit. Here we show that neither neurosteroid binds to the S1S2 domain of the NMDA NR1-1b subunit. This NR1-1b NMDA domain does, however, bind to the endogenous polyamines spermine and spermidine as well as Zn(II). Binding of the polyamines and Zn(II) to the S1S2 domain of the GluR2 subunit was not observed. This binding of Zn(II) and polyamines to the S1S2 domain of the NR1-1b subunit defines a new binding site for each of these modulators. PMID- 17114219 TI - Orientation and dynamics of melittin in membranes of varying composition utilizing NBD fluorescence. AB - Melittin is a cationic hemolytic peptide isolated from the European honey bee, Apis mellifera. The organization of membrane-bound melittin has earlier been shown to be dependent on the physical state and composition of membranes. In this study, we covalently labeled the N-terminal (Gly-1) and Lys-7 of melittin with an environment-sensitive fluorescent probe, the NBD group, to monitor the influence of negatively charged lipids and cholesterol on the organization and dynamics of membrane-bound melittin. Our results show that the NBD group of melittin labeled at its N-terminal end does not exhibit red edge excitation shift in DOPC and DOPC/DOPG membranes, whereas the NBD group of melittin labeled at Lys-7 exhibits REES of approximately 8 nm. This could be attributed to difference in membrane microenvironment experienced by the NBD groups in these analogs. Interestingly, the membrane environment of the NBD groups is sensitive to the presence of cholesterol, which is supported by time-resolved fluorescence measurements. Importantly, the orientation of melittin is found to be parallel to the membrane surface as determined by membrane penetration depth analysis using the parallax method in all cases. Our results constitute the first report to our knowledge describing the orientation of melittin in cholesterol-containing membranes. These results assume significance in the overall context of the role of membrane lipids in the orientation and function of membrane proteins and peptides. PMID- 17114220 TI - Insight into the putative specific interactions between cholesterol, sphingomyelin, and palmitoyl-oleoyl phosphatidylcholine. AB - The effects of cholesterol (Chol) on phospholipid bilayers include ordering of the fatty acyl chains, condensing of the lipids in the bilayer plane, and promotion of the liquid-ordered phase. These effects depend on the type of phospholipids in the bilayer and are determined by the nature of the underlying molecular interactions. As for Chol, it has been shown to interact more favorably with sphingomyelin than with most phosphatidylcholines, which in given circumstances leads to formation of lateral domains. However, the exact origin and nature of Chol-phospholipid interactions have recently been subjects of speculation. We examine interactions between Chol, palmitoylsphingomyelin (PSM) and palmitoyl-oleoyl-phosphatidylcholine (POPC) in hydrated lipid bilayers by extensive atom-scale molecular dynamics simulations. We employ a tailored lipid configuration: Individual PSM and Chol monomers, as well as PSM-Chol dimers, are embedded in a POPC lipid bilayer in the liquid crystalline phase. Such a setup allows direct comparison of dimeric and monomeric PSMs and Chol, which ultimately shows how the small differences in PSM and POPC structure can lead to profoundly different interactions with Chol. Our analysis shows that direct hydrogen bonding between PSM and Chol does not provide an adequate explanation for their putative specific interaction. Rather, a combination of charge-pairing, hydrophobic, and van der Waals interactions leads to a lower tilt in PSM neighboring Chol than in Chol with only POPC neighbors. This implies improved Chol-induced ordering of PSM's chains over POPC's chains. These findings are discussed in the context of the hydrophobic mismatch concept suggested recently. PMID- 17114221 TI - The cooperative response of synaptotagmin I C2A. A hypothesis for a Ca2+-driven molecular hammer. AB - In the current understanding of exocytosis at the nerve terminal, the C2 domain of synaptotagmin (C2A) is presumed to bind Ca2+ and the membrane in a stepwise fashion: cation then membrane as cation increases the affinity of protein for membrane. Fluorescence spectroscopy data were gathered over a variety of lipid and Ca2+ concentrations, enabling the rigorous application of microscopic binding models derived from partition functions to differentiate between Ca2+ and phosphatidylserine contributions to binding. The data presented here are in variance with previously published models, which were based on the Hill approximation. Rather, the data are consistent with two forms of cooperativity that modulate the responsiveness of C2A: in Ca2+ binding to a network of three cation sites and in interaction with the membrane surface. We suggest synaptotagmin I C2A is preassociated with the synaptic vesicle membrane or nerve terminal. In this state, upon Ca2+ influx the protein will bind the three Ca2+ ions immediately and with high cooperativity. Thus, membrane association creates a high-affinity Ca2+ switch that is the basis for the role of synaptotagmin I in Ca2+-regulated exocytosis. Based on this model, we discuss the implications of protein-induced phosphatidylserine demixing to the exocytotic process. PMID- 17114222 TI - Atomic-scale structure and electrostatics of anionic palmitoyloleoylphosphatidylglycerol lipid bilayers with Na+ counterions. AB - Anionic palmitoyloleoylphosphatidylglycerol (POPG) is one of the most abundant lipids in nature, yet its atomic-scale properties have not received significant attention. Here we report extensive 150-ns molecular dynamics simulations of a pure POPG lipid membrane with sodium counterions. It turns out that the average area per lipid of the POPG bilayer under physiological conditions is approximately 19% smaller than that of a bilayer built from its zwitterionic phosphatidylcholine analog, palmitoyloleoylphosphatidylcholine. This suggests that there are strong attractive interactions between anionic POPG lipids, which overcome the electrostatic repulsion between negative charges of PG headgroups. We demonstrate that interlipid counterion bridges and strong intra- and intermolecular hydrogen bonding play a key role in this seemingly counterintuitive behavior. In particular, the substantial strength and stability of ion-mediated binding between anionic lipid headgroups leads to complexation of PG molecules and ions and formation of large PG-ion clusters that act in a concerted manner. The ion-mediated binding seems to provide a possible molecular level explanation for the low permeability of PG-containing bacterial membranes to organic solvents: highly polar interactions at the water/membrane interface are able to create a high free energy barrier for hydrophobic molecules such as benzene. PMID- 17114223 TI - Coadsorption of human milk lactoferrin into the dipalmitoylglycerolphosphatidylcholine phospholipid monolayer spread at the air/water interface. AB - The coadsorption of human milk lactoferrin into a spread monolayer of dipalmitoylglycerol phosphatidylcholine (DPPC) at the air/water interface has been studied by neutron reflection. The system is a good model of the preocular tear film outer interface, which was the motivation for the study. The association of the protein with the surface was indicated by an increase of the surface pressure exerted by the DPPC monolayer. The extent of lactoferrin coadsorption was found to decrease with increasing surface pressure in the lipid monolayer, a trend consistent with the observation reported for other proteins, such as lysozyme and beta-lactoglobulin. The neutron reflectivity measurements were subsequently carried out at the three surface pressures of 8, 15, and 35 mN/m to examine the structure and composition of lactoferrin coadsorbed at the interface. Whereas the DPPC monolayer effectively prevented lactoferrin insertion at the high surface pressure, a measurable amount of lactoferrin was found at the air/water interface at the two lower surface pressures. At 15 mN/m it was difficult to identify the distribution of lactoferrin with respect to the DPPC monolayer, due to its relatively low adsorbed amount and much broader distribution. At the lowest surface pressure of 8 mN/m, the lactoferrin coadsorption was found to increase with time over the first few hours. After 5 h the distribution of the lactoferrin layer became similar to, though quantitatively lower than, that adsorbed in the absence of the DPPC monolayer. It is characterized by a top dense sublayer of 15 A with a bottom diffuse sublayer of 60 A, indicating structural unfolding induced by surface adsorption under these conditions. PMID- 17114224 TI - FRET study of membrane proteins: determination of the tilt and orientation of the N-terminal domain of M13 major coat protein. AB - A formalism for membrane protein structure determination was developed. This method is based on steady-state FRET data and information about the position of the fluorescence maxima on site-directed fluorescent labeled proteins in combination with global data analysis utilizing simulation-based fitting. The methodology was applied to determine the structural properties of the N-terminal domain of the major coat protein from bacteriophage M13 reconstituted into unilamellar DOPC/DOPG (4:1 mol/mol) vesicles. For our purpose, the cysteine mutants A7C, A9C, N12C, S13C, Q15C, A16C, S17C, and A18C in the N-terminal domain of this protein were produced and specifically labeled with the fluorescence probe AEDANS. The energy transfer data from the natural Trp-26 to AEDANS were analyzed assuming a two-helix protein model. Furthermore, the polarity Stokes shift of the AEDANS fluorescence maxima is taken into account. As a result the orientation and tilt of the N-terminal protein domain with respect to the bilayer interface were obtained, showing for the first time, to our knowledge, an overall alpha-helical protein conformation from amino acid residues 12-46, close to the protein conformation in the intact phage. PMID- 17114225 TI - Altered membrane dynamics of quantum dot-conjugated integrins during osteogenic differentiation of human bone marrow derived progenitor cells. AB - Functionalized quantum dots offer several advantages for tracking the motion of individual molecules on the cell surface, including selective binding, precise optical identification of cell surface molecules, and detailed examination of the molecular motion without photobleaching. We have used quantum dots conjugated with integrin antibodies and performed studies to quantitatively demonstrate changes in the integrin dynamics during osteogenic differentiation of human bone marrow derived progenitor cells (BMPCs). Consistent with the unusually strong BMPC adhesion previously observed, integrins on the surface of undifferentiated BMPC were found in clusters and the lateral diffusion was slow (e.g., approximately 10(-11) cm2/s). At times as early as those after a 3-day incubation in the osteogenic differentiation media, the integrin diffusion coefficients increased by an order of magnitude, and the integrin dynamics became indistinguishable from that measured on the surface of terminally differentiated human osteoblasts. Furthermore, microfilaments in BMPCs consisted of atypically thick bundles of stress fibers that were responsible for restricting the integrin lateral mobility. Studies using laser optical tweezers showed that, unlike fully differentiated osteoblasts, the BMPC cytoskeleton is weakly associated with its cell membrane. Based on these findings, it appears likely that the altered integrin dynamics is correlated with BMPC differentiation and that the integrin lateral mobility is restricted by direct links to microfilaments. PMID- 17114226 TI - Protein particulates: another generic form of protein aggregation? AB - Protein aggregation is a problem with a multitude of consequences, ranging from affecting protein expression to its implication in many diseases. Of recent interest is the specific form of aggregation leading to the formation of amyloid fibrils, structures associated with diseases such as Alzheimer's disease. The ability to form amyloid fibrils is now regarded as a property generic to all polypeptide chains. Here we show that around the isoelectric point a different generic form of aggregation can also occur by studying seven widely different, nonrelated proteins that are also all known to form amyloid fibrils. Under these conditions gels consisting of relatively monodisperse spherical particulates are formed. Although these gels have been described before for beta-lactoglobulin, our results suggest that the formation of particulates in the regime where charge on the molecules is minimal is a common property of all proteins. Because the proteins used here also form amyloid fibrils, we further propose that protein misfolding into clearly defined aggregates is a generic process whose outcome depends solely on the general properties of the state the protein is in when aggregation occurs, rather than the specific amino acid sequence. Thus under conditions of high net charge, amyloid fibrils form, whereas under conditions of low net charge, particulates form. This observation furthermore suggests that the rules of soft matter physics apply to these systems. PMID- 17114227 TI - Structural stabilization and functional improvement of horseradish peroxidase upon modification of accessible lysines: experiments and simulation. AB - Horseradish peroxidase (HRP) is an important heme enzyme with enormous medical diagnostic, biosensing, and biotechnological applications. Thus, any improvement in the applicability and stability of the enzyme is potentially interesting. We previously reported that covalent attachment of an electron relay (anthraquinone 2-carboxylic acid) to the surface-exposed Lys residues successfully improves electron transfer properties of HRP. Here we investigated structural and functional consequences of this modification, which alters three accessible charged lysines (Lys-174, Lys-232, and Lys-241) to the hydrophobic anthraquinolysine residues. Thermal denaturation and thermoinactivation studies demonstrated that this kind of modification enhances the conformational and operational stability of HRP. The melting temperature increased 3 degrees C and the catalytic efficiency enhanced by 80%. Fluorescence and circular dichroism investigations suggest that the modified HRP benefits from enhanced aromatic packing and more buried hydrophobic patches as compared to the native one. Molecular dynamics simulations showed that modification improves the accessibility of His-42 and the heme prosthetic group to the peroxide and aromatic substrates, respectively. Additionally, the hydrophobic patch, which functions as a binding site or trap for reducing aromatic substrates, is more extended in the modified enzyme. In summary, this modification produces a new derivative of HRP with enhanced electron transfer properties, catalytic efficiency, and stability for biotechnological applications. PMID- 17114228 TI - Dendritic spine viscoelasticity and soft-glassy nature: balancing dynamic remodeling with structural stability. AB - Neuronal dendritic spines are a key component of brain circuitry, implicated in many mechanisms for plasticity and long-term stability of synaptic communication. They can undergo rapid actin-based activity-dependent shape fluctuations, an intriguing biophysical property that is believed to alter synaptic transmission. Yet, because of their small size (approximately 1 microm or less) and metastable behavior, spines are inaccessible to most physical measurement techniques. Here we employ atomic force microscopy elasticity mapping and novel dynamic indentation methods to probe the biomechanics of dendritic spines in living neurons. We find that spines exhibit 1), a wide range of rigidities, correlated with morphological characteristics, axonal association, and glutamatergic stimulation, 2), a uniquely large viscosity, four to five times that of other cell types, consistent with a high density of solubilized proteins, and 3), weak power-law rheology, described by the soft-glassy model for cellular mechanics. Our findings provide a new perspective on spine functionality and identify key mechanical properties that govern the ability of spines to rapidly remodel and regulate internal protein trafficking but also maintain structural stability. PMID- 17114229 TI - The formation of fibrils by intertwining of filaments: model and application to amyloid Abeta protein. AB - We outline a model that describes the interaction of rods that form intertwined bundles. In this simple model, we compare the elastic energy penalty that arises due to the deformation of the rods to the gain in binding energy upon intertwining. We find that, for proper values of the bending Young's modulus and the binding energy, a helical pitch may be found for which the energy of intertwining is most favorable. We apply our description to the problem of Alzheimer's Abeta protein fibrillization. If we forbid configurations that exhibit steric overlap between the protofilaments that make up a protein fibril, our model predicts that fibrils consisting of three protofilaments shall form. This agrees well with experimental results. Our model can also provide an estimate for the helical pitch of suitable fibrils. PMID- 17114230 TI - Hydration changes accompanying the binding of minor groove ligands with DNA. AB - 4',6-diamidino-2-phenylindole (DAPI), netropsin, and pentamidine are minor groove binders that have terminal -C(NH2)2+ groups. The hydration changes that accompany their binding to the minor groove of the (AATT)2 sequence have been studied using the osmotic stress technique with fluorescence spectroscopy. The affinity of DAPI for the binding site decreases with the increasing osmolality of the solution, resulting in acquisition of 35+/-1 waters upon binding. A competition fluorescence assay was utilized to measure the binding constants and hydration changes of the other two ligands, using the DNA-DAPI complex as the fluorescence reporter. Upon their association to the (AATT)2 binding site, netropsin and pentamidine acquire 26+/-3 and 34+/-2 additional waters of hydration, respectively. The hydration changes are discussed in the context of the terminal functional groups of the ligands and conformational changes in the DNA. PMID- 17114231 TI - Membrane simulations of OpcA: gating in the loops? AB - Mobility of extracellular loops may play an important role in the function of outer membrane proteins from Gram-negative bacteria. Molecular dynamics simulations of OpcA from Neisseria meningitidis, embedded in a lipid bilayer, have been used to explore the relationship between the crystal structure and dynamic function of this protein. The results suggest that the crystal environment may constrain the membrane protein structure in a nonphysiological state. The presence of lipids and physiological salt concentrations result in changes in the conformation of the extracellular loops of OpcA, leading to opening of a pore, and to modulation of the molecular surface implicated in recognition of proteoglycan. These changes may be related to the role of OpcA in pathogenesis via modulation of the conformation of a possible sialic acid binding site. PMID- 17114232 TI - Bending elasticity of anti-parallel beta-sheets. AB - Using a coarse-grained elastic model, we examine the bending properties of anti parallel beta-sheets comprised of uniform amino-acid residues in vacuum as well as in explicit solvent. By comparing the conformational probability of the beta sheet from molecular dynamics simulations with the same quantities obtained from the coarse-grained model, we compute the elastic bending constant, kappa. Equilibrium fluctuations of the beta-sheet and its response to external forces are well reproduced by a model with a uniform isotropic bending constant. An anisotropic bending model is also investigated, although the computed anisotropy is relatively weak and most of the observed properties are well described by an isotropic model. The presence of explicit solvent also lowers the bending constant. The sequence dependence of our result and its implications in protein conformational dynamics are discussed. PMID- 17114233 TI - Ion conduction through MscS as determined by electrophysiology and simulation. AB - The mechanosensitive channel of small conductance (MscS) is a membrane protein thought to act as a safety valve in bacteria, regulating the release of ions and small solutes when gated by membrane tension under challenging osmotic conditions. The influence of voltage on channel activation and the functional state depicted by the available crystal structure of MscS remain debated. Therefore, in an effort to relate electrophysiological measurements on MscS and properties of the MscS crystal conformation, we report here MscS's response to voltage and pressure as determined by patch-clamp experiments, as well as MscS electrostatics and transport properties as determined through all-atom molecular dynamics simulations of the protein embedded in a lipid bilayer, a 224,000-atom system. The experiments reveal that MscS is a slightly anion-selective channel with a conductance of approximately 1 ns, activated by pressure and inactivated in a voltage-dependent manner. On the other hand, the simulations, covering over 200 ns and including biasing electrostatic potentials, show that MscS restrained to the crystal conformation exhibits low conductance; unrestrained it increases the channel radius upon application of a large electrostatic bias and exhibits then ion conduction that matches experimentally determined conductances. The simulated conductance stems mainly from Cl- ions. PMID- 17114234 TI - Cost-effectiveness of the unrestricted use of sirolimus-eluting stents vs. bare metal stents at 1 and 2-year follow-up: results from the RESEARCH Registry. AB - AIMS: To assess the cost-effectiveness of sirolimus-eluting stents (SESs) compared with bare metal stents (BMSs) as the default strategy in unselected patients treated in the Rapamycin Eluting Stent Evaluated At Rotterdam Cardiology Hospital (RESEARCH) Registry at 1 and 2-years following the procedure. METHODS AND RESULTS: A total of 508 consecutive patients with de novo lesions exclusively treated with SES were compared with 450 patients treated with BMS from the immediate preceding period. Resource use and costs of the index procedure, and clinical outcomes were prospectively recorded over a 2-year follow-up period. Follow-up costs were measured as unit costs per patient based on the incidence of clinically driven target vessel revascularization (TVR), to obtain cumulative costs at 1 and 2-years. Cost-effectiveness was measured as the incremental cost effectiveness ratio (ICER) per TVR avoided. The use of SES cost euro 3,036 more per patient at the index procedure, driven by the price of SES. Follow-up costs after 1-year were euro 1,089 less with SES when compared with BMS, due to less TVR, resulting in a net excess cost of euro 1,968 per patient in the SES group, and reduced by a further euro 100 per patient in the second year. The incidence of death or myocardial infarction between groups was similar at 1 and 2 years. Rates of TVR in the SES and BMS groups were 3.7% vs. 10.4%, P<0.01 at 1 year, respectively; and 6.4% vs. 14.7%, P<0.001 at 2 years. The ICER per TVR avoided was euro 29,373 at 1 year, and euro 22,267 at 2 years. CONCLUSION: The use of SES, while significantly beneficial in reducing the need for repeat revascularization, was more expensive and not cost-effective in the RESEARCH registry at either 1 or 2-years when compared with BMS. On the basis of these results, in an unselected population with 1 year of follow-up, the unit price of SES would have to be euro 1,023 in order to be cost-neutral. PMID- 17114235 TI - I2 imaging: cancer biology and the tumor microenvironment. AB - The use of imaging techniques to understand the role of the tumor microenvironment in cancer progression was the topic of a National Cancer Institute (NCI)-sponsored think tank entitled "I2 Imaging: Cancer Biology and the Tumor Microenvironment," held in Alexandria, Virginia on June 8 to 10, 2006. Participants discussed both recent progress in the use of imaging to dissect cellular and molecular interactions within the tumor microenvironment and the challenges that remain. Recommendations made to the NCI included (a) holding an annual meeting at which biologists, clinicians, and imaging scientists could exchange data, facilitating new collaborations within this multidisciplinary field; (b) funding both research and training specifically designed to foster a cross-disciplinary focus; (c) creating and making available a variety of resources to interested investigators, such as a repository of stromal cells and extracellular matrix molecules; and (d) taking steps to encourage translation of the basic research findings into the clinic. PMID- 17114236 TI - Marked genomic differences characterize primary and secondary glioblastoma subtypes and identify two distinct molecular and clinical secondary glioblastoma entities. AB - Glioblastoma is classified into two subtypes on the basis of clinical history: "primary glioblastoma" arising de novo without detectable antecedent disease and "secondary glioblastoma" evolving from a low-grade astrocytoma. Despite their distinctive clinical courses, they arrive at an indistinguishable clinical and pathologic end point highlighted by widespread invasion and resistance to therapy and, as such, are managed clinically as if they are one disease entity. Because the life history of a cancer cell is often reflected in the pattern of genomic alterations, we sought to determine whether primary and secondary glioblastomas evolve through similar or different molecular pathogenetic routes. Clinically annotated primary and secondary glioblastoma samples were subjected to high resolution copy number analysis using oligonucleotide-based array comparative genomic hybridization. Unsupervised classification using genomic nonnegative matrix factorization methods identified three distinct genomic subclasses. Whereas one corresponded to clinically defined primary glioblastomas, the remaining two stratified secondary glioblastoma into two genetically distinct cohorts. Thus, this global genomic analysis showed wide-scale differences between primary and secondary glioblastomas that were previously unappreciated, and has shown for the first time that secondary glioblastoma is heterogeneous in its molecular pathogenesis. Consistent with these findings, analysis of regional recurrent copy number alterations revealed many more events unique to these subclasses than shared. The pathobiological significance of these shared and subtype-specific copy number alterations is reinforced by their frequent occurrence, resident genes with clear links to cancer, recurrence in diverse cancer types, and apparent association with clinical outcome. We conclude that glioblastoma is composed of at least three distinct molecular subtypes, including novel subgroups of secondary glioblastoma, which may benefit from different therapeutic strategies. PMID- 17114237 TI - Macrophages regulate the angiogenic switch in a mouse model of breast cancer. AB - The development of a tumor vasculature or access to the host vasculature is a crucial step for the survival and metastasis of malignant tumors. Although therapeutic strategies attempting to inhibit this step during tumor development are being developed, the biological regulation of this process is still largely unknown. Using a transgenic mouse susceptible to mammary cancer, PyMT mice, we have characterized the development of the vasculature in mammary tumors during their progression to malignancy. We show that the onset of the angiogenic switch, identified as the formation of a high-density vessel network, is closely associated with the transition to malignancy. More importantly, both the angiogenic switch and the progression to malignancy are regulated by infiltrated macrophages in the primary mammary tumors. Inhibition of the macrophage infiltration into the tumor delayed the angiogenic switch and malignant transition whereas genetic restoration of the macrophage population specifically in these tumors rescued the vessel phenotype. Furthermore, premature induction of macrophage infiltration into premalignant lesions promoted an early onset of the angiogenic switch independent of tumor progression. Taken together, this study shows that tumor-associated macrophages play a key role in promoting tumor angiogenesis, an essential step in the tumor progression to malignancy. PMID- 17114238 TI - In vitro and in vivo activity of SKI-606, a novel Src-Abl inhibitor, against imatinib-resistant Bcr-Abl+ neoplastic cells. AB - Resistance to imatinib represents an important scientific and clinical issue in chronic myelogenous leukemia. In the present study, the effects of the novel inhibitor SKI-606 on various models of resistance to imatinib were studied. SKI 606 proved to be an active inhibitor of Bcr-Abl in several chronic myelogenous leukemia cell lines and transfectants, with IC(50) values in the low nanomolar range, 1 to 2 logs lower than those obtained with imatinib. Cells expressing activated forms of KIT or platelet-derived growth factor receptor (PDGFR), two additional targets of imatinib, were unaffected by SKI-606, whereas activity was found against PIM2. SKI-606 retained activity in cells where resistance to imatinib was caused by BCR-ABL gene amplification and in three of four Bcr-Abl point mutants tested. In vivo experiments confirmed SKI-606 activity in models where resistance was not caused by mutations as well as in cells carrying the Y253F, E255K, and D276G mutations. Modeling considerations attribute the superior activity of SKI-606 to its ability to bind a conformation of Bcr-Abl different from imatinib. PMID- 17114239 TI - Exercise-induced brachial artery vasodilation: role of free radicals. AB - Originally thought of as simply damaging or toxic "accidents" of in vivo chemistry, free radicals are becoming increasingly recognized as redox signaling molecules implicit in cellular homeostasis. Indeed, at the vascular level, it is plausible that oxidative stress plays a regulatory role in normal vascular function. Using electron paramagnetic resonance (EPR) spectroscopy, we sought to document the ability of an oral antioxidant cocktail (vitamins C, E, and alpha lipoic acid) to reduce circulating free radicals, and we employed Doppler ultrasound to examine the consequence of an antioxidant-mediated reduction in oxidative stress on exercise-induced vasodilation. A total of 25 young (18-31 yr) healthy male subjects partook in these studies. EPR spectroscopy revealed a reduction in circulating free radicals following antioxidant administration at rest ( approximately 98%) and as a consequence of exercise ( approximately 85%). Plasma total antioxidant capacity and vitamin C both increased following the ingestion of the antioxidant cocktail, whereas vitamin E levels were not influenced by the ingestion of the antioxidants. Brachial artery vasodilation during submaximal forearm handgrip exercise was greater with the placebo (7.4 +/- 1.8%) than with the antioxidant cocktail (2.3 +/- 0.7%). These data document the efficacy of an oral antioxidant cocktail in reducing free radicals and suggest that, in a healthy state, the aggressive disruption of the delicate balance between pro- and antioxidant forces can negatively impact vascular function. These findings implicate an exercise-induced reliance upon pro-oxidant-stimulated vasodilation, thereby revealing an important and positive vascular role for free radicals. PMID- 17114240 TI - High-fat diet postinfarction enhances mitochondrial function and does not exacerbate left ventricular dysfunction. AB - Lipid accumulation in nonadipose tissue due to enhanced circulating fatty acids may play a role in the pathophysiology of heart failure, obesity, and diabetes. Accumulation of myocardial lipids and related intermediates, e.g., ceramide, is associated with decreased contractile function, mitochondrial oxidative phosphorylation, and electron transport chain (ETC) complex activities. We tested the hypothesis that the progression of heart failure would be exacerbated by elevated myocardial lipids and an associated ceramide-induced inhibition of mitochondrial oxidative phosphorylation and ETC complex activities. Heart failure (HF) was induced by coronary artery ligation. Rats were then randomly assigned to either a normal (10% kcal from fat; HF, n = 8) or high saturated fat diet (60% kcal from saturated fat; HF + Sat, n = 7). Sham-operated animals (sham; n = 8) were fed a normal diet. Eight weeks postligation, left ventricular (LV) function was assessed by echocardiography and catheterization. Subsarcolemmal and interfibrillar mitochondria were isolated from the LV. Heart failure resulted in impaired LV contractile function [decreased percent fractional shortening and peak rate of LV pressure rise and fall (+/-dP/dt)] and remodeling (increased end diastolic and end-systolic dimensions) in HF compared with sham. No further progression of LV dysfunction was evident in HF + Sat. Mitochondrial state 3 respiration was increased in HF + Sat compared with HF despite elevated myocardial ceramide. Activities of ETC complexes II and IV were elevated in HF + Sat compared with HF and sham. High saturated fat feeding following coronary artery ligation was associated with increased oxidative phosphorylation and ETC complex activities and did not adversely affect LV contractile function or remodeling, despite elevations in myocardial ceramide. PMID- 17114241 TI - Magnesium attenuates isoproterenol-induced acute cardiac dysfunction and beta adrenergic desensitization. AB - Sympathetic nervous activation is a crucial compensatory mechanism in heart failure. However, excess catecholamine may induce cardiac dysfunction and beta adrenergic desensitization. Although magnesium is known to be a cardioprotective agent, its beneficial effects on acute cardiac dysfunction remain to be elucidated. We examined the effects of magnesium on left ventricular (LV) dysfunction induced by a large dose of isoproterenol in dogs. Sixteen anesthetized dogs underwent a continuous infusion of isoproterenol (1 micro g.kg( 1).min(-1)) with or without a magnesium infusion (1 mg.kg(-1).min(-1)). The dose response to small doses of isoproterenol (0.025-0.2 micro g.kg(-1).min(-1)) was tested hourly. A large dose of isoproterenol decreased LV systolic function, increased the time constant of LV isovolumic relaxation, and suppressed the dose response to small doses of isoproterenol in a time-dependent manner. Magnesium significantly attenuated isoproterenol-induced LV systolic and diastolic dysfunction and preserved the dose response to isoproterenol. Serum-ionized calcium significantly decreased with a large dose of isoproterenol but was fully maintained at baseline level with magnesium. A large dose of isoproterenol increased serum lipid peroxide levels and serological markers of myocardial damage, which were significantly suppressed by magnesium. In conclusion, magnesium significantly attenuated excess isoproterenol-induced acute cardiac dysfunction and beta-adrenergic desensitization. PMID- 17114242 TI - Wave intensity analysis of left atrial mechanics and energetics in anesthetized dogs. AB - The left atrium (LA) acts as a booster pump during late diastole, generating the Doppler transmitral A wave and contributing incrementally to left ventricular (LV) filling. However, after volume loading and in certain disease states, LA contraction fills the LV less effectively, and retrograde flow (i.e., the Doppler Ar wave) into the pulmonary veins increases. The purpose of this study was to provide an energetic analysis of LA contraction to clarify the mechanisms responsible for changes in forward and backward flow. Wave intensity analysis was performed at the mitral valve and a pulmonary vein orifice. As operative LV stiffness increased with progressive volume loading, the reflection coefficient (i.e., energy of reflected wave/energy of incident wave) also increased. This reflected wave decelerated the forward movement of blood through the mitral valve and was transmitted through the LA, accelerating retrograde blood flow in the pulmonary veins. Although total LA work increased with volume loading, the forward hydraulic work decreased and backward hydraulic work increased. Thus wave reflection due to increased LV stiffness accounts for the decrease in the A wave and the increase in the Ar wave measured by Doppler. PMID- 17114243 TI - High dietary salt reduces the contribution of 20-HETE to arteriolar oxygen responsiveness in skeletal muscle. AB - The coupling of tissue blood flow to cellular metabolic demand involves oxygen dependent adjustments in arteriolar tone, and arteriolar responses to oxygen can be mediated, in part, by changes in local production of 20-HETE. In this study, we examined the long-term effect of dietary salt on arteriolar oxygen responsiveness in the exteriorized, superfused rat spinotrapezius muscle and the role of 20-HETE in this responsiveness. Rats were fed either a normal-salt (NS, 0.45%) or high-salt (HS, 4%) diet for 4-5 wk. There was no difference in steady state tissue Po(2) between NS and HS rats, and elevation of superfusate oxygen content from 0% to 10% caused tissue Po(2) to increase by the same amount in both groups. However, the resulting reductions in arteriolar diameter and blood flow were less in HS rats than NS rats. Inhibition of 20-HETE formation with N methylsulfonyl-12,12-dibromododec-11-enamide (DDMS) or 17-octadecynoic acid (17 ODYA) attenuated oxygen-induced constriction in NS rats but not HS rats. Exogenous 20-HETE elicited arteriolar constriction that was greatly reduced by the large-conductance Ca(2+)-activated potassium (K(Ca)) channel inhibitors tetraethylammonium chloride (TEA) and iberiotoxin (IbTx) in NS rats and a smaller constriction that was less sensitive to TEA or IbTx in HS rats. Arteriolar responses to exogenous angiotensin II were similar in both groups but more sensitive to inhibition with DDMS in NS rats. Norepinephrine-induced arteriolar constriction was similar and insensitive to DDMS in both groups. We conclude that 20-HETE contributes to oxygen-induced constriction of skeletal muscle arterioles via inhibition of K(Ca) channels and that a high-salt diet impairs arteriolar responses to increased oxygen availability due to a reduction in vascular smooth muscle responsiveness to 20-HETE. PMID- 17114244 TI - Ca2+ signaling in mouse mesenteric small arteries: myogenic tone and adrenergic vasoconstriction. AB - Arteries that have developed myogenic tone (MT) are in a markedly different physiological state compared with those that have not, with higher cytosolic [Ca(2+)] and altered activity of several signal transduction pathways. In this study, we sought to determine whether alpha(1)-adrenoceptor-induced Ca(2+) signaling is different in pressurized arteries that have spontaneously developed MT (the presumptive physiological state) compared with those that have not (a common experimental state). At 32 degrees C and intraluminal pressure of 70 mmHg, cytoplasmic [Ca(2+)] was steady in most smooth muscle cells (SMCs). In a minority of cells (34%), however, at least one propagating Ca(2+) wave occurred. alpha(1) Adrenoceptor activation (phenylephrine, PE; 0.1-10.0 microM) caused strong vasoconstriction and markedly increased the frequency of Ca(2+) waves (in virtually all cells). However, when cytosolic [Ca(2+)] was elevated experimentally in these arteries ([K(+)] 20 mM), PE failed to elicit Ca(2+) waves, although it did elevate [Ca(2+)] (F/F(0)) further and caused further vasoconstriction. During development of MT, the cytosolic [Ca(2+)] (F/F(0)) in individual SMCs increased, Ca(2+) waves disappeared (from SMCs that had them), and small Ca(2+) ripples (frequency approximately 0.05 Hz) appeared in approximately 13% of cells. PE elicited only spatially uniform increases in [Ca(2+)] and a smaller change in diameter (than in the absence of MT). Nevertheless, when cytosolic [Ca(2+)] and MT were decreased by nifedipine (1 microM), PE did elicit Ca(2+) waves. Thus alpha(1)-adrenoceptor-mediated Ca(2+) signaling is markedly different in arteries with and without MT, perhaps due to the elevated [Ca(2+)], and may have a different molecular basis. alpha(1) Adrenoceptor-induced vasoconstriction may be supported either by Ca(2+) waves or by steady elevation of cytoplasmic [Ca(2+)], depending on the amount of MT. PMID- 17114245 TI - Endothelial nitric oxide synthase (NOS3) knockout decreases NOS2 induction, limiting hyperoxygenation and conferring protection in the postischemic heart. AB - Although it has been shown that endothelial nitric oxide synthase (eNOS)-derived nitric oxide downregulates mitochondrial oxygen consumption during early reperfusion, its effects on inducible NOS (iNOS) induction and myocardial injury during late reperfusion are unknown. Wild-type (WT) and eNOS(-/-) mice were subjected to 30 min of coronary ligation followed by reperfusion. Expression of iNOS mRNA and protein levels and peroxynitrite production were lower in postischemic myocardium of eNOS(-/-) mice than levels in WT mice 48 h postreperfusion. Significantly improved hemodynamics (+/-dP/dt, left ventricular systolic pressure, mean arterial pressure), increased rate pressure product, and reduced myocardial infarct size (18 +/- 2.5% vs. 31 +/- 4.6%) were found 48 h after reperfusion in eNOS(-/-) mice compared with WT mice. Myocardial infarct size was also significantly decreased in WT mice treated with the specific iNOS inhibitor 1400W (20.5 +/- 3.4%) compared with WT mice treated with PBS (33.9 +/- 5.3%). A marked reperfusion-induced hyperoxygenation state was observed by electron paramagnetic resonance oximetry in postischemic myocardium, but Po(2) values were significantly lower from 1 to 72 h in eNOS(-/-) than in WT mice. Cytochrome c-oxidase activity and NADH dehydrogenase activity were significantly decreased in postischemic myocardium in WT and eNOS(-/-) mice compared with baseline control, respectively, and NADH dehydrogenase activity was significantly higher in eNOS(-/-) than in WT mice. Thus deficiency of eNOS exerted a sustained beneficial effect on postischemic myocardium 48 h after reperfusion with preserved mitochondrial function, which appears to be due to decreased iNOS induction and decreased iNOS-derived peroxynitrite in postischemic myocardium. PMID- 17114246 TI - Deficiency of TNFR1 protects myocardium through SOCS3 and IL-6 but not p38 MAPK or IL-1beta. AB - Tumor necrosis factor-alpha (TNF-alpha) plays an important role in the development of heart failure. There is a direct correlation between myocardial function and myocardial TNF levels in humans. TNF may induce local inflammation to exert tissue injury. On the other hand, suppressors of cytokine signaling (SOCS) proteins have been shown to inhibit proinflammatory signaling. However, it is unknown whether TNF mediates myocardial inflammation via STAT3/SOCS3 signaling in the heart and, if so, whether this effect is through the type 1 55-kDa TNF receptor (TNFR1). We hypothesized that TNFR1 deficiency protects myocardial function and decreases myocardial IL-6 production via the STAT3/SOCS3 pathway in response to TNF. Isolated male mouse hearts (n = 4/group) from wild-type (WT) and TNFR1 knockout (TNFR1KO) were subjected to direct TNF infusion (500 pg.ml( 1).min(-1) x 30 min) while left ventricular developed pressure and maximal positive and negative values of the first derivative of pressure were continuously recorded. Heart tissue was analyzed for active forms of STAT3, p38, SOCS3 and SOCS1 (Western blot analysis), as well as IL-1beta and IL-6 (ELISA). Coronary effluent was analyzed for lactate dehydrogenase (LDH) activity. As a result, TNFR1KO had significantly better myocardial function, less myocardial LDH release, and greater expression of SOCS3 (percentage of SOCS3/GAPDH: 45 +/- 4.5% vs. WT 22 +/- 6.5%) after TNF infusion. TNFR1 deficiency decreased STAT3 activation (percentage of phospho-STAT3/STAT3: 29 +/- 6.4% vs. WT 45 +/- 8.8%). IL-6 was decreased in TNFR1KO (150.2 +/- 3.65 pg/mg protein) versus WT (211.4 +/- 26.08) mice. TNFR1 deficiency did not change expression of p38 and IL-1beta following TNF infusion. These results suggest that deficiency of TNFR1 protects myocardium through SOCS3 and IL-6 but not p38 MAPK or IL-1beta. PMID- 17114247 TI - Derepression of excision of integrative and potentially conjugative elements from Streptococcus thermophilus by DNA damage response: implication of a cI-related repressor. AB - A DNA-damaging agent, mitomycin C, derepresses the site-specific excision of two integrative and potentially conjugative elements from Streptococcus thermophilus, ICESt1 and ICESt3. The regulation pathway involves a repressor related to phage lambda cI repressor. It could also involve a putative regulator related to another type of phage repressors, the "cI-like" repressors. PMID- 17114248 TI - The Streptococcus mutans vicX gene product modulates gtfB/C expression, biofilm formation, genetic competence, and oxidative stress tolerance. AB - Streptococcus mutans is considered one of the primary etiologic agents of dental caries. Previously, we characterized the VicRK two-component signal transduction system, which regulates multiple virulence factors of S. mutans. In this study, we focused on the vicX gene of the vicRKX tricistronic operon. To characterize vicX, we constructed a nonpolar deletion mutation in the vicX coding region in S. mutans UA159. The growth kinetics of the mutant (designated SmuvicX) showed that the doubling time was longer and that there was considerable sensitivity to paraquat-induced oxidative stress. Supplementing a culture of the wild-type UA159 strain with paraquat significantly increased the expression of vicX (P < 0.05, as determined by analysis of variance [ANOVA]), confirming the role of this gene in oxidative stress tolerance in S. mutans. Examination of mutant biofilms revealed architecturally altered cell clusters that were seemingly denser than the wild type cell clusters. Interestingly, vicX-deficient cells grown in a glucose supplemented medium exhibited significantly increased glucosyltransferase B/C (gtfB/C) expression compared with the expression in the wild type (P < 0.05, as determined by ANOVA). Moreover, a sucrose-dependent adhesion assay performed using an S. mutans GS5-derived vicX null mutant demonstrated that the adhesiveness of this mutant was enhanced compared with that of the parent strain and isogenic mutants of the parent strain lacking gtfB and/or gtfC. Also, disruption of vicX reduced the genetic transformability of the mutant approximately 10-fold compared with that of the parent strain (P < 0.05, as determined by ANOVA). Collectively, these findings provide insight into important phenotypes controlled by the vicX gene product that can impact S. mutans pathogenicity. PMID- 17114249 TI - Role of a putative polysaccharide locus in Bordetella biofilm development. AB - Bordetellae are gram-negative bacteria that colonize the respiratory tracts of animals and humans. We and others have recently shown that these bacteria are capable of living as sessile communities known as biofilms on a number of abiotic surfaces. During the biofilm mode of existence, bacteria produce one or more extracellular polymeric substances that function, in part, to hold the cells together and to a surface. There is little information on either the constituents of the biofilm matrix or the genetic basis of biofilm development by Bordetella spp. By utilizing immunoblot assays and by enzymatic hydrolysis using dispersin B (DspB), a glycosyl hydrolase that specifically cleaves the polysaccharide poly beta-1,6-N-acetyl-D-glucosamine (poly-beta-1,6-GlcNAc), we provide evidence for the production of poly-beta-1,6-GlcNAc by various Bordetella species (Bordetella bronchiseptica, B. pertussis, and B. parapertussis) and its role in their biofilm development. We have investigated the role of a Bordetella locus, here designated bpsABCD, in biofilm formation. The bps (Bordetella polysaccharide) locus is homologous to several bacterial loci that are required for the production of poly beta-1,6-GlcNAc and have been implicated in bacterial biofilm formation. By utilizing multiple microscopic techniques to analyze biofilm formation under both static and hydrodynamic conditions, we demonstrate that the bps locus, although not essential at the initial stages of biofilm formation, contributes to the stability and the maintenance of the complex architecture of Bordetella biofilms. PMID- 17114250 TI - Physical and functional interactions between Escherichia coli MutY glycosylase and mismatch repair protein MutS. AB - Escherichia coli MutY and MutS increase replication fidelity by removing adenines that were misincorporated opposite 7,8-dihydro-8-oxo-deoxyguanines (8-oxoG), G, or C. MutY DNA glycosylase removes adenines from these mismatches through a short patch base excision repair pathway and thus prevents G:C-to-T:A and A:T-to-G:C mutations. MutS binds to the mismatches and initiates the long-patch mismatch repair on daughter DNA strands. We have previously reported that the human MutY homolog (hMYH) physically and functionally interacts with the human MutS homolog, hMutSalpha (Y. Gu et al., J. Biol. Chem. 277:11135-11142, 2002). Here, we show that a similar relationship between MutY and MutS exists in E. coli. The interaction of MutY and MutS involves the Fe-S domain of MutY and the ATPase domain of MutS. MutS, in eightfold molar excess over MutY, can enhance the binding activity of MutY with an A/8-oxoG mismatch by eightfold. The MutY expression level and activity in mutS mutant strains are sixfold and twofold greater, respectively, than those for the wild-type cells. The frequency of A:T to-G:C mutations is reduced by two- to threefold in a mutS mutY mutant compared to a mutS mutant. Our results suggest that MutY base excision repair and mismatch repair defend against the mutagenic effect of 8-oxoG lesions in a cooperative manner. PMID- 17114251 TI - RamB, the transcriptional regulator of acetate metabolism in Corynebacterium glutamicum, is subject to regulation by RamA and RamB. AB - In Corynebacterium glutamicum, the transcriptional regulator RamB negatively controls the expression of genes involved in acetate metabolism. Here we show that RamB represses its own expression by direct interaction with a 13-bp motif in the ramB promoter region. Additionally, ramB expression is subject to carbon source-dependent positive control by RamA. PMID- 17114252 TI - Biological relevance of colony morphology and phenotypic switching by Burkholderia pseudomallei. AB - Melioidosis is a notoriously protracted illness and is difficult to cure. We hypothesize that the causative organism, Burkholderia pseudomallei, undergoes a process of adaptation involving altered expression of surface determinants which facilitates persistence in vivo and that this is reflected by changes in colony morphology. A colony morphotyping scheme and typing algorithm were developed using clinical B. pseudomallei isolates. Morphotypes were divided into seven types (denoted I to VII). Type I gave rise to other morphotypes (most commonly type II or III) by a process of switching in response to environmental stress, including starvation, iron limitation, and growth at 42 degrees C. Switching was associated with complex shifts in phenotype, one of which (type I to type II) was associated with a marked increase in production of factors putatively associated with in vivo concealment. Isogenic types II and III, derived from type I, were examined using several experimental models. Switching between isogenic morphotypes occurred in a mouse model, where type II appeared to become adapted for persistence in a low-virulence state. Isogenic type II demonstrated a significant increase in intracellular replication fitness compared with parental type I after uptake by epithelial cells in vitro. Isogenic type III demonstrated a higher replication fitness following uptake by macrophages in vitro, which was associated with a switch to type II. Mixed B. pseudomallei morphologies were common in individual clinical specimens and were significantly more frequent in samples of blood, pus, and respiratory secretions than in urine and surface swabs. These findings have major implications for therapeutics and vaccine development. PMID- 17114253 TI - Role of the beta1 subunit in the function and stability of the 20S proteasome in the hyperthermophilic archaeon Pyrococcus furiosus. AB - The hyperthermophilic archaeon Pyrococcus furiosus genome encodes three proteasome component proteins: one alpha protein (PF1571) and two beta proteins (beta1-PF1404 and beta2-PF0159), as well as an ATPase (PF0115), referred to as proteasome-activating nucleotidase. Transcriptional analysis of the P. furiosus dynamic heat shock response (shift from 90 to 105 degrees C) showed that the beta1 gene was up-regulated over twofold within 5 minutes, suggesting a specific role during thermal stress. Consistent with transcriptional data, two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that incorporation of the beta1 protein relative to beta2 into the 20S proteasome (core particle [CP]) increased with increasing temperature for both native and recombinant versions. For the recombinant enzyme, the beta2/beta1 ratio varied linearly with temperature from 3.8, when assembled at 80 degrees C, to 0.9 at 105 degrees C. The recombinant alpha+beta1+beta2 CP assembled at 105 degrees C was more thermostable than either the alpha+beta1+beta2 version assembled at 90 degrees C or the alpha+beta2 version assembled at either 90 degrees C or 105 degrees C, based on melting temperature and the biocatalytic inactivation rate at 115 degrees C. The recombinant CP assembled at 105 degrees C was also found to have different catalytic rates and specificity for peptide hydrolysis, compared to the 90 degrees C assembly (measured at 95 degrees C). Combination of the alpha and beta1 proteins neither yielded a large proteasome complex nor demonstrated any significant activity. These results indicate that the beta1 subunit in the P. furiosus 20S proteasome plays a thermostabilizing role and influences biocatalytic properties, suggesting that beta subunit composition is a factor in archaeal proteasome function during thermal stress, when polypeptide turnover is essential to cell survival. PMID- 17114254 TI - Essential bacterial functions encoded by gene pairs. AB - To address the need for new antibacterials, a number of bacterial genomes have been systematically disrupted to identify essential genes. Such programs have focused on the disruption of single genes and may have missed functions encoded by gene pairs or multiple genes. In this work, we hypothesized that we could predict the identity of pairs of proteins within one organism that have the same function. We identified 135 putative protein pairs in Bacillus subtilis and attempted to disrupt the genes forming these, singly and then in pairs. The single gene disruptions revealed new genes that could not be disrupted individually and other genes required for growth in minimal medium or for sporulation. The pairwise disruptions revealed seven pairs of proteins that are likely to have the same function, as the presence of one protein can compensate for the absence of the other. Six of these pairs are essential for bacterial viability and in four cases show a pattern of species conservation appropriate for potential antibacterial development. This work highlights the importance of combinatorial studies in understanding gene duplication and identifying functional redundancy. PMID- 17114255 TI - Identification of diverse archaeal proteins with class III signal peptides cleaved by distinct archaeal prepilin peptidases. AB - Most secreted archaeal proteins are targeted to the membrane via a tripartite signal composed of a charged N terminus and a hydrophobic domain, followed by a signal peptidase-processing site. Signal peptides of archaeal flagellins, similar to class III signal peptides of bacterial type IV pilins, are distinct in that their processing sites precede the hydrophobic domain, which is crucial for assembly of these extracytoplasmic structures. To identify the complement of archaeal proteins with class III signal sequences, a PERL program (FlaFind) was written. A diverse set of proteins was identified, and many of these FlaFind positives were encoded by genes that were cotranscribed with homologs of pilus assembly genes. Moreover, structural conservation of primary sequences between many FlaFind positives and subunits of bacterial pilus-like structures, which have been shown to be critical for pilin assembly, have been observed. A subset of pilin-like FlaFind positives contained a conserved domain of unknown function (DUF361) within the signal peptide. Many of the genes encoding these proteins were in operons that contained a gene encoding a novel euryarchaeal prepilin peptidase, EppA, homolog. Heterologous analysis revealed that Methanococcus maripaludis DUF361-containing proteins were specifically processed by the EppA homolog of this archaeon. Conversely, M. maripaludis preflagellins were cleaved only by the archaeal preflagellin peptidase FlaK. Together, the results reveal a diverse set of archaeal proteins with class III signal peptides that might be subunits of as-yet-undescribed cell surface structures, such as archaeal pili. PMID- 17114256 TI - Genetic diversity among Botulinum Neurotoxin-producing clostridial strains. AB - Clostridium botulinum is a taxonomic designation for many diverse anaerobic spore forming rod-shaped bacteria that have the common property of producing botulinum neurotoxins (BoNTs). The BoNTs are exoneurotoxins that can cause severe paralysis and death in humans and other animal species. A collection of 174 C. botulinum strains was examined by amplified fragment length polymorphism (AFLP) analysis and by sequencing of the 16S rRNA gene and BoNT genes to examine the genetic diversity within this species. This collection contained representatives of each of the seven different serotypes of botulinum neurotoxins (BoNT/A to BoNT/G). Analysis of the16S rRNA gene sequences confirmed previous identifications of at least four distinct genomic backgrounds (groups I to IV), each of which has independently acquired one or more BoNT genes through horizontal gene transfer. AFLP analysis provided higher resolution and could be used to further subdivide the four groups into subgroups. Sequencing of the BoNT genes from multiple strains of serotypes A, B, and E confirmed significant sequence variation within each serotype. Four distinct lineages within each of the BoNT A and B serotypes and five distinct lineages of serotype E strains were identified. The nucleotide sequences of the seven toxin genes of the serotypes were compared and showed various degrees of interrelatedness and recombination, as was previously noted for the nontoxic nonhemagglutinin gene, which is linked to the BoNT gene. These analyses contribute to the understanding of the evolution and phylogeny within this species and assist in the development of improved diagnostics and therapeutics for the treatment of botulism. PMID- 17114257 TI - Morphogenesis of the Bacillus anthracis spore. AB - Bacillus spp. and Clostridium spp. form a specialized cell type, called a spore, during a multistep differentiation process that is initiated in response to starvation. Spores are protected by a morphologically complex protein coat. The Bacillus anthracis coat is of particular interest because the spore is the infective particle of anthrax. We determined the roles of several B. anthracis orthologues of Bacillus subtilis coat protein genes in spore assembly and virulence. One of these, cotE, has a striking function in B. anthracis: it guides the assembly of the exosporium, an outer structure encasing B. anthracis but not B. subtilis spores. However, CotE has only a modest role in coat protein assembly, in contrast to the B. subtilis orthologue. cotE mutant spores are fully virulent in animal models, indicating that the exosporium is dispensable for infection, at least in the context of a cotE mutation. This has implications for both the pathophysiology of the disease and next-generation therapeutics. CotH, which directs the assembly of an important subset of coat proteins in B. subtilis, also directs coat protein deposition in B. anthracis. Additionally, however, in B. anthracis, CotH effects germination; in its absence, more spores germinate than in the wild type. We also found that SpoIVA has a critical role in directing the assembly of the coat and exosporium to an area around the forespore. This function is very similar to that of the B. subtilis orthologue, which directs the assembly of the coat to the forespore. These results show that while B. anthracis and B. subtilis rely on a core of conserved morphogenetic proteins to guide coat formation, these proteins may also be important for species-specific differences in coat morphology. We further hypothesize that variations in conserved morphogenetic coat proteins may play roles in taxonomic variation among species. PMID- 17114258 TI - Making a point: the role of DivIVA in streptococcal polar anatomy. PMID- 17114259 TI - Evolution of catabolic pathways: Genomic insights into microbial s-triazine metabolism. PMID- 17114260 TI - Functional reconstitution of SdcS, a Na+-coupled dicarboxylate carrier protein from Staphylococcus aureus. AB - In Staphylococcus aureus, the transport of dicarboxylates is mediated in part by the Na+-linked carrier protein SdcS. This transporter is a member of the divalent anion/Na+ symporter (DASS) family, a group that includes the mammalian Na+/dicarboxylate cotransporters NaDC1 and NaDC3. In earlier work, we cloned and expressed SdcS in Escherichia coli and found it to have transport properties similar to those of its eukaryotic counterparts (J. A. Hall and A. M. Pajor, J. Bacteriol. 187:5189-5194, 2005). Here, we report the partial purification and subsequent reconstitution of functional SdcS into liposomes. These proteoliposomes exhibited succinate counterflow activity, as well as Na+ electrochemical-gradient-driven transport. Examination of substrate specificity indicated that the minimal requirement necessary for transport was a four-carbon terminal dicarboxylate backbone and that productive substrate-transporter interaction was sensitive to substitutions at the substrate C-2 and C-3 positions. Further analysis established that SdcS facilitates an electroneutral symport reaction having a 2:1 cation/dicarboxylate ratio. This study represents the first characterization of a reconstituted Na+-coupled DASS family member, thus providing an effective method to evaluate functional, as well as structural, aspects of DASS transporters in a system free of the complexities and constraints associated with native membrane environments. PMID- 17114261 TI - Characterization of OpdH, a Pseudomonas aeruginosa porin involved in the uptake of tricarboxylates. AB - The Pseudomonas aeruginosa outer membrane is intrinsically impermeable to many classes of antibiotics, due in part to its relative lack of general uptake pathways. Instead, this organism relies on a large number of substrate-specific uptake porins. Included in this group are the 19 members of the OprD family, which are involved in the uptake of a diverse array of metabolites. One of these porins, OpdH, has been implicated in the uptake of cis-aconitate. Here we demonstrate that this porin may also enable P. aeruginosa to take up other tricarboxylates. Isocitrate and citrate strongly and specifically induced the opdH gene via a mechanism involving derepression by the putative two-component regulatory system PA0756-PA0757. Planar bilayer analysis of purified OpdH demonstrated that it was a channel-forming protein with a large single-channel conductance (230 pS in 1 M KCl; 10-fold higher than that of OprD); however, we were unable to demonstrate the presence of a tricarboxylate binding site within the channel. Thus, these data suggest that the requirement for OpdH for efficient growth on tricarboxylates was likely due to the specific expression of this large channel porin under particular growth conditions. PMID- 17114262 TI - Catalase (KatA) and alkyl hydroperoxide reductase (AhpC) have compensatory roles in peroxide stress resistance and are required for survival, persistence, and nasal colonization in Staphylococcus aureus. AB - Oxidative-stress resistance in Staphylococcus aureus is linked to metal ion homeostasis via several interacting regulators. In particular, PerR controls the expression of a regulon of genes, many of which encode antioxidants. Two PerR regulon members, ahpC (alkylhydroperoxide reductase) and katA (catalase), show compensatory regulation, with independent and linked functions. An ahpC mutation leads to increased H2O2 resistance due to greater katA expression via relief of PerR repression. Moreover, AhpC provides residual catalase activity present in a katA mutant. Mutation of both katA and ahpC leads to a severe growth defect under aerobic conditions in defined media (attributable to lack of catalase activity). This results in the inability to scavenge exogenous or endogenously produced H2O2, resulting in accumulation of H2O2 in the medium. This leads to DNA damage, the likely cause of the growth defect. Surprisingly, the katA ahpC mutant is not attenuated in two independent models of infection, which implies reduced oxygen availability during infection. In contrast, both AhpC and KatA are required for environmental persistence (desiccation) and nasal colonization. Thus, oxidative stress resistance is an important factor in the ability of S. aureus to persist in the hospital environment and so contribute to the spread of human disease. PMID- 17114263 TI - Translation control of trpG from transcripts originating from the folate operon promoter of Bacillus subtilis is influenced by translation-mediated displacement of bound TRAP, while translation control of transcripts originating from a newly identified trpG promoter is not. AB - Bacillus subtilis trpG encodes a glutamine amidotransferase subunit that participates in the biosynthesis of both tryptophan and folic acid. TRAP inhibits translation of trpG in response to tryptophan by binding to a site that overlaps the trpG Shine-Dalgarno sequence, thereby blocking ribosome binding. Similar mechanisms regulate trpP and ycbK translation. The equilibrium binding constants of tryptophan-activated TRAP for the trpG, ycbK, and trpP transcripts were determined to be 8, 3, and 50 nM, respectively. Despite TRAP having a higher affinity for the trpG transcript, TRAP exhibited the least control of trpG expression. The trpG Shine-Dalgarno sequence overlaps the stop codon of the upstream pabB gene, while six of nine triplet repeats within the TRAP binding site are located upstream of the pabB stop codon. Thus, ribosomes translating the upstream pabB cistron could be capable of reducing TRAP-dependent control of TrpG synthesis by displacing bound TRAP. Expression studies using pabB-trpG'-'lacZ fusions in the presence or absence of an engineered stop codon within pabB suggest that translation-mediated displacement of bound TRAP reduces TRAP dependent inhibition of TrpG synthesis from transcripts originating from the folate operon promoter (P(pabB)). A new trpG promoter (P(trpG)) was identified in the pabB coding sequence that makes a larger contribution to trpG expression than does P(pabB). We found that TRAP-dependent regulation of trpG expression is more extensive for a transcript originating from P(trpG) and that transcripts originating from P(trpG) are not subject to translation-mediated displacement of bound TRAP. PMID- 17114264 TI - Pathway confirmation and flux analysis of central metabolic pathways in Desulfovibrio vulgaris hildenborough using gas chromatography-mass spectrometry and Fourier transform-ion cyclotron resonance mass spectrometry. AB - Flux distribution in central metabolic pathways of Desulfovibrio vulgaris Hildenborough was examined using 13C tracer experiments. Consistent with the current genome annotation and independent evidence from enzyme activity assays, the isotopomer results from both gas chromatography-mass spectrometry (GC-MS) and Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICR MS) indicate the lack of an oxidatively functional tricarboxylic acid (TCA) cycle and an incomplete pentose phosphate pathway. Results from this study suggest that fluxes through both pathways are limited to biosynthesis. The data also indicate that >80% of the lactate was converted to acetate and that the reactions involved are the primary route of energy production [NAD(P)H and ATP production]. Independently of the TCA cycle, direct cleavage of acetyl coenzyme A to CO and 5,10-methyl tetrahydrofuran also leads to production of NADH and ATP. Although the genome annotation implicates a ferredoxin-dependent oxoglutarate synthase, isotopic evidence does not support flux through this reaction in either the oxidative or the reductive mode; therefore, the TCA cycle is incomplete. FT-ICR MS was used to locate the labeled carbon distribution in aspartate and glutamate and confirmed the presence of an atypical enzyme for citrate formation suggested in previous reports [the citrate synthesized by this enzyme is the isotopic antipode of the citrate synthesized by the (S)-citrate synthase]. These findings enable a better understanding of the relation between genome annotation and actual metabolic pathways in D. vulgaris and also demonstrate that FT-ICR MS is a powerful tool for isotopomer analysis, overcoming the problems with both GC-MS and nuclear magnetic resonance spectroscopy. PMID- 17114265 TI - Bacillus subtilis genome diversity. AB - Microarray-based comparative genomic hybridization (M-CGH) is a powerful method for rapidly identifying regions of genome diversity among closely related organisms. We used M-CGH to examine the genome diversity of 17 strains belonging to the nonpathogenic species Bacillus subtilis. Our M-CGH results indicate that there is considerable genetic heterogeneity among members of this species; nearly one-third of Bsu168-specific genes exhibited variability, as measured by the microarray hybridization intensities. The variable loci include those encoding proteins involved in antibiotic production, cell wall synthesis, sporulation, and germination. The diversity in these genes may reflect this organism's ability to survive in diverse natural settings. PMID- 17114266 TI - Characterization of Francisella tularensis outer membrane proteins. AB - Francisella tularensis is a gram-negative coccobacillus that is capable of causing severe, fatal disease in a number of mammalian species, including humans. Little is known about the proteins that are surface exposed on the outer membrane (OM) of F. tularensis, yet identification of such proteins is potentially fundamental to understanding the initial infection process, intracellular survival, virulence, immune evasion and, ultimately, vaccine development. To facilitate the identification of putative F. tularensis outer membrane proteins (OMPs), the genomes of both the type A strain (Schu S4) and type B strain (LVS) were subjected to six bioinformatic analyses for OMP signatures. Compilation of the bioinformatic predictions highlighted 16 putative OMPs, which were cloned and expressed for the generation of polyclonal antisera. Total membranes were extracted from both Schu S4 and LVS by spheroplasting and osmotic lysis, followed by sucrose density gradient centrifugation, which separated OMs from cytoplasmic (inner) membrane and other cellular compartments. Validation of OM separation and enrichment was confirmed by probing sucrose gradient fractions with antibodies to putative OMPs and inner membrane proteins. F. tularensis OMs typically migrated in sucrose gradients between densities of 1.17 and 1.20 g/ml, which differed from densities typically observed for other gram-negative bacteria (1.21 to 1.24 g/ml). Finally, the identities of immunogenic proteins were determined by separation on two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometric analysis. This is the first report of a direct method for F. tularensis OM isolation that, in combination with computational predictions, offers a more comprehensive approach for the characterization of F. tularensis OMPs. PMID- 17114267 TI - Genetic characterization and virulence role of the RALP3/LSA locus upstream of the streptolysin s operon in invasive M1T1 Group A Streptococcus. AB - Group A Streptococcus (GAS) is a leading human pathogen associated with a wide spectrum of mucosal and invasive infections. GAS expresses a large number of virulence determinants whose expression is under the control of several transcriptional regulatory networks. Here we performed the first mutational analysis of a genetic locus immediately upstream of the streptolysin S biosynthetic operon in several GAS genome sequences, including that of the M1T1 serotype, the leading isolates associated with serious invasive disease. The locus consists of a predicted RofA-like stand-alone transcriptional regulator (RALP3) and the largest open reading frame in the GAS genome, encoding a predicted LPXSG motif cell wall-anchored protein we have named LSA (for "large surface-anchored" protein). Comparative reverse transcription-PCR analysis of wild-type M1T1 GAS and an isogenic RALP3-deficient mutant identifies RALP3 as a global transcriptional regulator affecting expression of numerous virulence factor genes, including those for strong repression of the hyaluronic acid capsule and cysteine protease production. RALP3 contributed to GAS epithelial cell invasion and bloodstream survival. LSA was found to be under negative regulation by RALP3 and to influence GAS-epithelial cell interactions and GAS antimicrobial peptide sensitivity. Isogenic M1T1 GAS mutants lacking either RALP3 or LSA were attenuated in a murine model of systemic infection, indicating that this locus plays a role in the virulence potential of the organism. PMID- 17114268 TI - Anaerobic central metabolic pathways in Shewanella oneidensis MR-1 reinterpreted in the light of isotopic metabolite labeling. AB - It has been proposed that during growth under anaerobic or oxygen-limited conditions, Shewanella oneidensis MR-1 uses the serine-isocitrate lyase pathway common to many methylotrophic anaerobes, in which formaldehyde produced from pyruvate is condensed with glycine to form serine. The serine is then transformed through hydroxypyruvate and glycerate to enter central metabolism at phosphoglycerate. To examine its use of the serine-isocitrate lyase pathway under anaerobic conditions, we grew S. oneidensis MR-1 on [1-13C]lactate as the sole carbon source, with either trimethylamine N-oxide (TMAO) or fumarate as an electron acceptor. Analysis of cellular metabolites indicated that a large percentage (>70%) of lactate was partially oxidized to either acetate or pyruvate. The 13C isotope distributions in amino acids and other key metabolites indicate that under anaerobic conditions, although glyoxylate synthesized from the isocitrate lyase reaction can be converted to glycine, a complete serine isocitrate pathway is not present and serine/glycine is, in fact, oxidized via a highly reversible degradation pathway. The labeling data also suggest significant activity in the anapleurotic (malic enzyme and phosphoenolpyruvate carboxylase) reactions. Although the tricarboxylic acid (TCA) cycle is often observed to be incomplete in many other anaerobes (absence of 2-oxoglutarate dehydrogenase activity), isotopic labeling supports the existence of a complete TCA cycle in S. oneidensis MR-1 under certain anaerobic conditions, e.g., TMAO-reducing conditions. PMID- 17114269 TI - AccD6, a member of the Fas II locus, is a functional carboxyltransferase subunit of the acyl-coenzyme A carboxylase in Mycobacterium tuberculosis. AB - The Mycobacterium tuberculosis acyl-coenzyme A (CoA) carboxylases provide the building blocks for de novo fatty acid biosynthesis by fatty acid synthase I (FAS I) and for the elongation of FAS I end products by the FAS II complex to produce meromycolic acids. The M. tuberculosis genome contains three biotin carboxylase subunits (AccA1 to -3) and six carboxyltransferase subunits (AccD1 to -6), with accD6 located in a genetic locus that contains members of the FAS II complex. We found by quantitative real-time PCR analysis that the transcripts of accA3, accD4, accD5, and accD6 are expressed at high levels during the exponential growth phases of M. tuberculosis in vitro. Microarray analysis of M. tuberculosis transcripts indicated that the transcripts for accA3, accD4, accD5, accD6, and accE were repressed during later growth stages. AccD4 and AccD5 have been previously studied, but there are no reports on the function of AccD6. We expressed AccA3 (alpha3) and AccD6 (beta6) in E. coli and purified them by affinity chromatography. We report here that reconstitution of the alpha3-beta6 complex yielded an active acyl-CoA carboxylase. Kinetic characterization of this carboxylase showed that it preferentially carboxylated acetyl-CoA (1.1 nmol/mg/min) over propionyl-CoA (0.36 nmol/mg/min). The activity of the alpha3 beta6 complex was inhibited by the epsilon subunit. The alpha3-beta6 carboxylase was inhibited significantly by dimethyl itaconate, C75, haloxyfop, cerulenin, and 1,2-cyclohexanedione. Our results suggest that the beta6 subunit could play an important role in mycolic acid biosynthesis by providing malonyl-CoA to the FAS II complex. PMID- 17114271 TI - Tobacco isoenzyme 1 of NAD(H)-dependent glutamate dehydrogenase catabolizes glutamate in vivo. AB - Glutamate (Glu) dehydrogenase (GDH, EC 1.4.1.2-1.4.1.4) catalyzes in vitro the reversible amination of 2-oxoglutarate to Glu. The in vivo direction(s) of the GDH reaction in higher plants and hence the role(s) of this enzyme is unclear, a situation confounded by the existence of isoenzymes comprised totally of either GDH beta- (isoenzyme 1) or alpha- (isoenzyme 7) subunits, as well as another five alpha-beta isoenzyme permutations. To clarify the in vivo direction of the reaction catalyzed by GDH isoenzyme 1, [(15)N]Glu was supplied to roots of two independent transgenic tobacco (Nicotiana tabacum) lines with increased isoenzyme 1 levels (S4-H and S49-H). The [(15)N]ammonium (NH(4)(+)) accumulation rate in these lines was elevated approximately 65% compared with a null segregant control line, indicating that isoenzyme 1 catabolizes Glu in roots. Leaf glutamine synthetase (GS) was inhibited with a GS-specific herbicide to quantify any contribution by GDH toward photorespiratory NH(4)(+) reassimilation. Transgenic line S49-H did not show enhanced resistance to the herbicide, indicating that the large pool of isoenzyme 1 in S49-H leaves was unable to compensate for GS and suggesting that isoenzyme 1 does not assimilate NH(4)(+) in vivo. PMID- 17114270 TI - Insights into the role of specific lipids in the formation and delivery of lipid microdomains to the plasma membrane of plant cells. AB - The existence of sphingolipid- and sterol-enriched microdomains, known as lipid rafts, in the plasma membrane (PM) of eukaryotic cells is well documented. To obtain more insight into the lipid molecular species required for the formation of microdomains in plants, we have isolated detergent (Triton X-100)-resistant membranes (DRMs) from the PM of Arabidopsis (Arabidopsis thaliana) and leek (Allium porrum) seedlings as well as from Arabidopsis cell cultures. Here, we show that all DRM preparations are enriched in sterols, sterylglucosides, and glucosylceramides (GluCer) and depleted in glycerophospholipids. The GluCer of DRMs from leek seedlings contain hydroxypalmitic acid. We investigated the role of sterols in DRM formation along the secretory pathway in leek seedlings. We present evidence for the presence of DRMs in both the PM and the Golgi apparatus but not in the endoplasmic reticulum. In leek seedlings treated with fenpropimorph, a sterol biosynthesis inhibitor, the usual Delta(5)-sterols are replaced by 9beta,19-cyclopropylsterols. In these plants, sterols and hydroxypalmitic acid-containing GluCer do not reach the PM, and most DRMs are recovered from the Golgi apparatus, indicating that Delta(5)-sterols and GluCer play a crucial role in lipid microdomain formation and delivery to the PM. In addition, DRM formation in Arabidopsis cells is shown to depend on the unsaturation degree of fatty acyl chains as evidenced by the dramatic decrease in the amount of DRMs prepared from the Arabidopsis mutants, fad2 and Fad3+, affected in their fatty acid desaturases. PMID- 17114272 TI - Cold transiently activates calcium-permeable channels in Arabidopsis mesophyll cells. AB - Living organisms are capable of discriminating thermal stimuli from noxious cold to noxious heat. For more than 30 years, it has been known that plant cells respond to cold with a large and transient depolarization. Recently, using transgenic Arabidopsis (Arabidopsis thaliana) expressing the calcium-sensitive protein aequorin, an increase in cytosolic calcium following cold treatment was observed. Applying the patch-clamp technique to Arabidopsis mesophyll protoplasts, we could identify a transient plasma membrane conductance induced by rapid cooling. This cold-induced transient conductance was characterized as an outward rectifying 33 pS nonselective cation channel. The permeability ratio between calcium and cesium was 0.7, pointing to a permeation pore >3.34 A (o of cesium). Our experiments thus provide direct evidence for the predicted but not yet measured cold-activated calcium-permeable channel in plants. PMID- 17114273 TI - Short vegetative phase-like MADS-box genes inhibit floral meristem identity in barley. AB - Analysis of the functions of Short Vegetative Phase (SVP)-like MADS-box genes in barley (Hordeum vulgare) indicated a role in determining meristem identity. Three SVP-like genes are expressed in vegetative tissues of barley: Barley MADS1 (BM1), BM10, and Vegetative to Reproductive Transition gene 2. These genes are induced by cold but are repressed during floral development. Ectopic expression of BM1 inhibited spike development and caused floral reversion in barley, with florets at the base of the spike replaced by tillers. Head emergence was delayed in plants that ectopically express BM1, primarily by delayed development after the floral transition, but expression levels of the barley VRN1 gene (HvVRN1) were not affected. Ectopic expression of BM10 inhibited spike development and caused partial floral reversion, where florets at the base of the spike were replaced by inflorescence-like structures, but did not affect heading date. Floral reversion occurred more frequently when BM1 and BM10 ectopic expression lines were grown in short-day conditions. BM1 and BM10 also inhibited floral development and caused floral reversion when expressed in Arabidopsis (Arabidopsis thaliana). We conclude that SVP-like genes function to suppress floral meristem identity in winter cereals. PMID- 17114274 TI - Putative role of aquaporins in variable hydraulic conductance of leaves in response to light. AB - Molecular and physiological studies in walnut (Juglans regia) are combined to establish the putative role of leaf plasma membrane aquaporins in the response of leaf hydraulic conductance (K(leaf)) to irradiance. The effects of light and temperature on K(leaf) are described. Under dark conditions, K(leaf) was low, but increased by 400% upon exposure to light. In contrast to dark conditions, K(leaf) values of light-exposed leaves responded to temperature and 0.1 mm cycloheximide treatments. Furthermore, K(leaf) was not related to stomatal aperture. Data of real-time reverse transcription-polymerase chain reaction showed that K(leaf) dynamics were tightly correlated with the transcript abundance of two walnut aquaporins (JrPIP2,1 and JrPIP2,2). Low K(leaf) in the dark was associated with down-regulation, whereas high K(leaf) in the light was associated with up regulation of JrPIP2. Light responses of K(leaf) and aquaporin transcripts were reversible and inhibited by cycloheximide, indicating the importance of de novo protein biosynthesis in this process. Our results indicate that walnut leaves can rapidly change their hydraulic conductance and suggest that these changes can be explained by regulation of plasma membrane aquaporins. Model simulation suggests that variable leaf hydraulic conductance in walnut might enhance leaf gas exchanges while buffering leaf water status in response to ambient light fluctuations. PMID- 17114275 TI - Decreases in stomatal conductance of soybean under open-air elevation of [CO2] are closely coupled with decreases in ecosystem evapotranspiration. AB - Stomatal responses to atmospheric change have been well documented through a range of laboratory- and field-based experiments. Increases in atmospheric concentration of CO(2) ([CO(2)]) have been shown to decrease stomatal conductance (g(s)) for a wide range of species under numerous conditions. Less well understood, however, is the extent to which leaf-level responses translate to changes in ecosystem evapotranspiration (ET). Since many changes at the soil, plant, and canopy microclimate levels may feed back on ET, it is not certain that a decrease in g(s) will decrease ET in rain-fed crops. To examine the scaling of the effect of elevated [CO(2)] on g(s) at the leaf to ecosystem ET, soybean (Glycine max) was grown in field conditions under control (approximately 375 micromol CO(2) mol(-1) air) and elevated [CO(2)] (approximately 550 micromol mol( 1)) using free air CO(2) enrichment. ET was determined from the time of canopy closure to crop senescence using a residual energy balance approach over four growing seasons. Elevated [CO(2)] caused ET to decrease between 9% and 16% depending on year and despite large increases in photosynthesis and seed yield. Ecosystem ET was linked with g(s) of the upper canopy leaves when averaged across the growing seasons, such that a 10% decrease in g(s) results in a 8.6% decrease in ET; this relationship was not altered by growth at elevated [CO(2)]. The findings are consistent with model and historical analyses that suggest that, despite system feedbacks, decreased g(s) of upper canopy leaves at elevated [CO(2)] results in decreased transfer of water vapor to the atmosphere. PMID- 17114276 TI - The mechanical diversity of stomata and its significance in gas-exchange control. AB - Given that stomatal movement is ultimately a mechanical process and that stomata are morphologically and mechanically diverse, we explored the influence of stomatal mechanical diversity on leaf gas exchange and considered some of the constraints. Mechanical measurements were conducted on the guard cells of four different species exhibiting different stomatal morphologies, including three variants on the classical "kidney" form and one "dumb-bell" type; this information, together with gas-exchange measurements, was used to model and compare their respective operational characteristics. Based on evidence from scanning electron microscope images of cryo-sectioned leaves that were sampled under full sun and high humidity and from pressure probe measurements of the stomatal aperture versus guard cell turgor relationship at maximum and zero epidermal turgor, it was concluded that maximum stomatal apertures (and maximum leaf diffusive conductance) could not be obtained in at least one of the species (the grass Triticum aestivum) without a substantial reduction in subsidiary cell osmotic (and hence turgor) pressure during stomatal opening to overcome the large mechanical advantage of subsidiary cells. A mechanism for this is proposed, with a corollary being greatly accelerated stomatal opening and closure. Gas-exchange measurements on T. aestivum revealed the capability of very rapid stomatal movements, which may be explained by the unique morphology and mechanics of its dumb-bell-shaped stomata coupled with "see-sawing" of osmotic and turgor pressure between guard and subsidiary cells during stomatal opening or closure. Such properties might underlie the success of grasses. PMID- 17114277 TI - Tobacco nectaries express a novel NADPH oxidase implicated in the defense of floral reproductive tissues against microorganisms. AB - Hydrogen peroxide produced from the nectar redox cycle was shown to be a major factor contributing to inhibition of most microbial growth in floral nectar; however, this obstacle can be overcome by the floral pathogen Erwinia amylovora. To identify the source of superoxide that leads to hydrogen peroxide accumulation in nectary tissues, nectaries were stained with nitroblue tetrazolium. Superoxide production was localized near nectary pores and inhibited by diphenylene iodonium but not by cyanide or azide, suggesting that NAD(P)H oxidase is the source of superoxide. Native PAGE assays demonstrated that NADPH (not NADH) was capable of driving the production of superoxide, diphenyleneiodonium chloride was an efficient inhibitor of this activity, but cyanide and azide did not inhibit. These results confirm that the production of superoxide was due to an NADPH oxidase. The nectary enzyme complex was distinct by migration on gels from the leaf enzyme complex. Temporal expression patterns demonstrated that the superoxide production (NADPH oxidase activity) was coordinated with nectar secretion, the expression of Nectarin I (a superoxide dismutase in nectar), and the expression of NOX1, a putative gene for a nectary NADPH oxidase that was cloned from nectaries and identified as an rbohD-like NADPH oxidase. Further, in situ hybridization studies indicated that the NADPH oxidase was expressed in the early stages of flower development although superoxide was generated at later stages (after Stage 10), implicating posttranslational regulation of the NADPH oxidase in the nectary. PMID- 17114278 TI - AtERF14, a member of the ERF family of transcription factors, plays a nonredundant role in plant defense. AB - We had previously shown that several transcription factors of the ethylene (ET) response factor (ERF) family were induced with different but overlapping kinetics following challenge of Arabidopsis (Arabidopsis thaliana) with Pseudomonas syringae pv tomato DC3000 (avrRpt2). One of these genes, a transcriptional activator, AtERF14, was induced at the same time as ERF-target genes (ChiB, basic chitinase). To unravel the potential function of AtERF14 in regulating the plant defense response, we have analyzed gain- and loss-of-function mutants. We show here that AtERF14 has a prominent role in the plant defense response, since overexpression of AtERF14 had dramatic effects on both plant phenotype and defense gene expression and AtERF14 loss-of-function mutants showed impaired induction of defense genes following exogenous ET treatment and increased susceptibility to Fusarium oxysporum. Moreover, the expression of other ERF genes involved in defense and ET/jasmonic acid responses, such as ERF1 and AtERF2, depends on AtERF14 expression. A number of ERFs have been shown to function in the defense response through overexpression. However, the effect of loss of AtERF14 function on defense gene expression, pathogen resistance, and regulation of the expression of other ERF genes is unique thus far. These results suggest a unique role for AtERF14 in regulating the plant defense response. PMID- 17114279 TI - Morphometry of the extremely thin pulmonary blood-gas barrier in the chicken lung. AB - The gas exchanging region in the avian lung, although proportionally smaller than that of the mammalian lung, efficiently manages respiration to meet the high energetic requirements of flapping flight. Gas exchange in the bird lung is enhanced, in part, by an extremely thin blood-gas barrier (BGB). We measured the arithmetic mean thickness of the different components (endothelium, interstitium, and epithelium) of the BGB in the domestic chicken lung and compared the results with three mammals. Morphometric analysis showed that the total BGB of the chicken lung was significantly thinner than that of the rabbit, dog, and horse (54, 66, and 70% thinner, respectively) and that all layers of the BGB were significantly thinner in the chicken compared with the mammals. The interstitial layer was strikingly thin in the chicken lung ( approximately 86% thinner than the dog and horse, and 75% thinner than rabbit) which is a paradox because the strength of the BGB is believed to come from the interstitium. In addition, the thickness of the interstitium was remarkably uniform, unlike the mammalian interstitium. The uniformity of the interstitial layer in the chicken is attributable to a lack of the supportive type I collagen cable that is found in mammalian alveolar lungs. We propose that the surrounding air capillaries provide additional structural support for the pulmonary capillaries in the bird lung, thus allowing the barrier to be both very thin and extremely uniform. The net result is to improve gas exchanging efficiency. PMID- 17114280 TI - Acid aspiration-induced lung inflammation and injury are exacerbated in NADPH oxidase-deficient mice. AB - Increased reactive oxidant intermediates (ROIs) from primed leukocytes have been implicated in the pathogenesis of acid aspiration lung injury. To evaluate the specific role of the phagocyte NADPH oxidase-derived ROIs in acid lung injury, the p47phox-/- knockout mouse model of chronic granulomatous disease was used. p47phox-/- mice developed a significantly greater alveolar neutrophilic leukocytosis compared with wild-type mice at all time points after acid injury, with the difference between genotypes being most marked at 48 h. In contrast, the p47phox-/- mice had a decreased number of macrophages in bronchoalveolar lavage (BAL) compared with wild-type at 48 h after acid or saline aspiration. Albumin concentration in BAL reflecting capillary leak was also greater in p47phox-/- compared with wild-type mice. BAL concentrations of proinflammatory cytokines and chemokines were greater in p47phox-/- compared with wild-type mice. These findings suggest that NADPH oxidase, directly or indirectly, plays a role in attenuating the acute neutrophilic response after acid lung injury. We speculate that this downmodulating effect may be mediated by promoting the transition from production of cytokines and chemokines involved in neutrophilic infiltration to a less injurious, chronic inflammatory response. PMID- 17114281 TI - Mesenchymal maintenance of distal epithelial cell phenotype during late fetal lung development. AB - Classical tissue recombination experiments have reported that at early gestation both tracheal and distal lung epithelium have the plasticity to respond to mesenchymal signals. Herein we examined the role of epithelial-mesenchymal interactions in maintaining epithelial differentiation at late (E19-E21, term = 22 days) fetal gestation in the rat. Isolated distal lung epithelial cells were recombined with mesenchymal cells from lung, skin, and intestine, and the homotypic or heterotypic recombinant cell aggregates were cultured for up to 5 days. Recombining lung epithelial cells with mesenchyme from various sources induced a morphological pattern that was specific to the type of inducing mesenchyme. In situ analysis of surfactant protein (SP)-C, SP-B, and Clara cell secretory protein (CCSP) expression, as well as SP-C and CCSP promoter transactivation experiments, revealed that distal lung epithelium requires lung mesenchyme to maintain the alveolar, but not bronchiolar, phenotype. Incubation of lung recombinants with an anti-FGF7 antibody resulted in a partial inhibition of mesenchyme-induced SP-C promoter transactivation. Immunoreactivity for Delta and Lunatic fringe, components of the Notch pathway that regulates cell differentiation, was downregulated in the heterotypic recombinants. In contrast, Hes1 mRNA expression was increased in these recombinants. Cumulatively, these results suggest that at late fetal gestation, distal lung epithelial cells are not fully committed to a specific phenotype and still have the plasticity to respond to various signals. Their alveolar phenotype is likely maintained by Notch/Notch ligand interactions and mesenchymal factors, including FGF7. PMID- 17114282 TI - Soluble guanylate cyclase-dependent relaxation is reduced in the adult rat bronchial smooth muscle. AB - Cyclic nucleotides are relaxants of the airway smooth muscle, yet most of the available data were obtained in adult animals. The expression and activity of cyclases have been reported to be developmentally regulated in the lung, and little is known about the age-related changes in their bronchial muscle relaxation potential. We evaluated and compared the newborn and adult rat bronchial smooth muscle response to cyclic AMP- and GMP-dependent agonists in isometric mounted bronchial rings. In acetylcholine-precontracted bronchial muscle, the relaxant response to the cAMP agonist forskolin was not age dependent, but the relaxant response to the nitric oxide (NO) donor sodium nitroprusside (SNP) was significantly greater (P<0.01) in the newborn. To further evaluate the cGMP pathway, we stimulated the soluble guanylate cyclase (sGC) with the specific agonists BAY 41-2272 and YC-1. In keeping with the SNP dose-response curves, the sGC agonists significantly relaxed the newborn, but not the adult bronchial muscle. Protein expression of the sGC alpha1- and beta1-subunits were significantly lower (P<0.01) in the adult compared with the newborn bronchial tissue. Consistent with these results, the NO-stimulated sGC activity was significantly greater in the newborn compared with the adult (P<0.01). In conclusion, the bronchial smooth muscle cGMP-, but not cAMP-dependent, relaxant response is developmentally regulated and significantly reduced in the adult rat. PMID- 17114283 TI - Bone morphogenetic protein signaling regulates the size of hair follicles and modulates the expression of cell cycle-associated genes. AB - Bone morphogenetic protein (BMP) signaling is involved in the regulation of a large variety of developmental programs, including those controlling organ sizes. Here, we show that transgenic (TG) mice overexpressing the BMP antagonist noggin (promoter, K5) are characterized by a marked increase in size of anagen hair follicles (HFs) and by the replacement of zig-zag and auchen hairs by awl-like hairs, compared with the age-matched WT controls. Markedly enlarged anagen HFs of TG mice show increased proliferation in the matrix and an increased number of hair cortex and medulla cells compared with WT HFs. Microarray and real-time PCR analyses of the laser-captured hair matrix cells show a strong decrease in expression of Cdk inhibitor p27(Kip1) and increased expression of selected cyclins in TG vs. WT mice. Similar to TG mice, p27(Kip1) knockout mice also show an increased size of anagen HFs associated with increased cell proliferation in the hair bulb. Primary epidermal keratinocytes (KC) from TG mice exhibit significantly increased proliferation and decreased p27(Kip1) expression, compared with WT KC. Alternatively, activation of BMP signaling in HaCaT KC induces growth arrest, stimulates p27(Kip1) expression, and positively regulates p27(Kip1) promoter activity, thus further supporting a role of p27(Kip1) in mediating the effects of BMP signaling on HF size. These data suggest that BMP signaling plays an important role in regulating cell proliferation and controls the size of anagen HFs by modulating the expression of cell-cycle-associated genes in hair matrix KC. PMID- 17114284 TI - Characterization of a cancer/testis (CT) antigen gene family capable of eliciting humoral response in cancer patients. AB - Cancer/testis (CT) antigens are immunogenic proteins expressed in normal gametogenic tissues and in different types of tumors. CT antigens are promising candidates for cancer immunotherapy, and the identification of novel CT antigens is a prerequisite for the development of cancer vaccines. We have identified a CT antigen, named CTSP-1, with partial similarity to the breast differentiation antigen NY-BR-1. CTSP-1 presents several splicing and polyadenylation variants and has a very restricted expression pattern among normal tissues. CTSP-1 is exclusively expressed in normal testis and is aberrantly expressed in 47.6% (10 of 21) of tumor cell lines and in 44.4% (75 of 169) of tumors from different histological types. The highest percentages of positive expression were observed in melanomas (59.0%) followed by prostate (58.0%) and lung (57.0%) tumors. CTSP-1 is part of a highly conserved gene family, and members of this family also have a restricted expression pattern and similar protein structure. Antibodies against members of this gene family were detected in 10% (14 of 141) of plasma samples from patients with a wide spectrum of tumors. The highest percentages of antibody response were observed in patients with prostate (20.8%), thyroid (20.0%), and breast (16.6%) tumors. Because of its very restricted expression pattern in normal tissues and immunogenicity in different types of tumors, CTSP-1 should be considered a promising candidate for cancer immunotherapy. PMID- 17114285 TI - The gatekeeper residue controls autoactivation of ERK2 via a pathway of intramolecular connectivity. AB - Studies of protein kinases have identified a "gatekeeper" residue, which confers selectivity for binding nucleotides and small-molecule inhibitors. We report that, in the MAP kinase ERK2, mutations at the gatekeeper residue unexpectedly lead to autoactivation due to enhanced autophosphorylation of regulatory Tyr and Thr sites within the activation lip that control kinase activity. This occurs through an intramolecular mechanism, indicating that the gatekeeper residue indirectly constrains flexibility at the activation lip, precluding access of the phosphoacceptor residues to the catalytic base. Other residues that interact with the gatekeeper site to form a hydrophobic cluster in the N-terminal domain also cause autoactivation when mutated. Hydrogen-exchange studies of a mutant within this cluster reveal perturbations in the conserved DFG motif, predicting a route for side chain connectivity from the hydrophobic cluster to the activation lip. Mutations of residues along this route support this model, explaining how information about the gatekeeper residue identity can be transmitted to the activation lip. Thus, an N-terminal hydrophobic cluster that includes the gatekeeper forms a novel structural unit, which functions to maintain the "off" state of ERK2 before cell signal activation. PMID- 17114286 TI - Bone marrow-derived fibroblast precursors mediate ischemic cardiomyopathy in mice. AB - We previously described a mouse model of fibrotic ischemia/reperfusion cardiomyopathy (I/RC) arising from daily, brief coronary occlusion. One characteristic of I/RC was the prolonged elevation of monocyte chemoattractant protein 1 (MCP-1), which was obligate to its phenotype and may contribute to the uptake of bloodborne cells. Here we describe in I/RC hearts a population of small spindle-shaped fibroblasts that were highly proliferative and expressed collagen I and alpha-smooth muscle actin (myofibroblast markers), CD34 (a precursor marker), and CD45 (a hematopoietic marker). These cells represented 3% of all nonmyocyte live cells. To confirm the cells' bone marrow origin, chimeric mice were created by the rescue of irradiated C57BL/6 mice with marrow from ROSA26, a congenic line expressing lacZ. I/RC resulted in a large population of spindle shaped fibroblasts containing lacZ. We postulated that the fibroblast precursors represented a developmental path for a subset of monocytes, whose phenotype we have shown to be influenced by serum amyloid P (SAP). Thus, we administered SAP in vivo, which markedly reduced the number of proliferative spindle-shaped fibroblasts and completely prevented I/RC-induced fibrosis and global ventricular dysfunction. By contrast, SAP did not suppress the inflammation or chemokine expression seen in I/RC. SAP, a member of the pentraxin family, binds to Fcgamma receptors and modifies the pathophysiological function of monocytes. Our data suggest that SAP interferes with assumption of a fibroblast phenotype in a subset of monocytes and that SAP may be an important regulator in the linkage between inflammation and nonadaptive fibrosis in the heart. PMID- 17114287 TI - Laser trapping in anisotropic fluids and polarization-controlled particle dynamics. AB - Anisotropic fluids are widespread, ranging from liquid crystals used in displays to ordered states of a biological cell interior. Optical trapping is potentially a powerful technique in the fundamental studies and applications of anisotropic fluids. We demonstrate that laser beams in these fluids can generate anisotropic optical trapping forces, even for particles larger than the trapping beam wavelength. Immersed colloidal particles modify the fluid's ordered molecular structures and locally distort its optic axis. This distortion produces a refractive index "corona" around the particles that depends on their surface characteristics. The laser beam can trap such particles not only at their center but also at the high-index corona. Trapping forces in the beam's lateral plane mimic the corona and are polarization-controlled. This control allows the optical forces to be reversed and cause the particle to follow a prescribed trajectory. Anisotropic particle dynamics in the trap varies with laser power because of the anisotropy of both viscous drag and trapping forces. Using thermotropic liquid crystals and biological materials, we show that these phenomena are quite general for all anisotropic fluids and impinge broadly on their quantitative studies using laser tweezers. Potential applications include modeling thermodynamic systems with anisotropic polarization-controlled potential wells, producing optically tunable photonic crystals, and fabricating light-controlled nano- and micropumps. PMID- 17114288 TI - Suppression of spontaneous genome rearrangements in yeast DNA helicase mutants. AB - Saccharomyces cerevisiae mutants lacking two of the three DNA helicases Sgs1, Srs2, and Rrm3 exhibit slow growth that is suppressed by disrupting homologous recombination. Cells lacking Sgs1 and Rrm3 accumulate gross-chromosomal rearrangements (GCRs) that are suppressed by the DNA damage checkpoint and by homologous recombination-defective mutations. In contrast, rrm3, srs2, and srs2 rrm3 mutants have wild-type GCR rates. GCR types in helicase double mutants include telomere additions, translocations, and broken DNAs healed by a complex process of hairpin-mediated inversion. Spontaneous activation of the Rad53 checkpoint kinase in the rrm3 mutant depends on the Mec3/Rad24 DNA damage sensors and results from activation of the Mec1/Rad9-dependent DNA damage response rather than the Mrc1-dependent replication stress response. Moreover, helicase double mutants accumulate Rad51-dependent Ddc2 foci, indicating the presence of recombination intermediates that are sensed by checkpoints. These findings demonstrate that different nonreplicative helicases function at the interface between replication and repair to maintain genome integrity. PMID- 17114289 TI - Genomic analysis of the uncultivated marine crenarchaeote Cenarchaeum symbiosum. AB - Crenarchaeota are ubiquitous and abundant microbial constituents of soils, sediments, lakes, and ocean waters. To further describe the cosmopolitan nonthermophilic Crenarchaeota, we analyzed the genome sequence of one representative, the uncultivated sponge symbiont Cenarchaeum symbiosum. C. symbiosum genotypes coinhabiting the same host partitioned into two dominant populations, corresponding to previously described a- and b-type ribosomal RNA variants. Although they were syntenic, overlapping a- and b-type ribotype genomes harbored significant variability. A single tiling path comprising the dominant a type genotype was assembled and used to explore the genomic properties of C. symbiosum and its planktonic relatives. Of 2,066 ORFs, 55.6% matched genes with predicted function from previously sequenced genomes. The remaining genes partitioned between functional RNAs (2.4%) and hypotheticals (42%) with limited homology to known functional genes. The latter category included some genes likely involved in the archaeal-sponge symbiotic association. Conversely, 525 C. symbiosum ORFs were most highly similar to sequences from marine environmental genomic surveys, and they apparently represent orthologous genes from free-living planktonic Crenarchaeota. In total, the C. symbiosum genome was remarkably distinct from those of other known Archaea and shared many core metabolic features in common with its free-living planktonic relatives. PMID- 17114290 TI - Increased prevalence of insulin resistance and nonalcoholic fatty liver disease in Asian-Indian men. AB - Type 2 diabetes mellitus (T2DM) is strongly associated with obesity in most, but not all, ethnic groups, suggesting important ethnic differences in disease susceptibility. Although it is clear that insulin resistance plays a major role in the pathogenesis of T2DM and that insulin resistance is strongly associated with increases in hepatic (HTG) and/or intramyocellular lipid content, little is known about the prevalence of insulin resistance and potential differences in intracellular lipid distribution among healthy, young, lean individuals of different ethnic groups. To examine this question, 482 young, lean, healthy, sedentary, nonsmoking Eastern Asians (n = 49), Asian-Indians (n = 59), Blacks (n = 48), Caucasians (n = 292), and Hispanics (n = 34) underwent an oral glucose tolerance test to assess whole-body insulin sensitivity by an insulin sensitivity index. In addition, intramyocellular lipid and HTG contents were measured by using proton magnetic resonance spectroscopy. The prevalence of insulin resistance, defined as the lower quartile of insulin sensitivity index, was approximately 2- to 3-fold higher in the Asian-Indians compared with all other ethnic groups, and this could entirely be attributed to a 3- to 4-fold increased prevalence of insulin resistance in Asian-Indian men. This increased prevalence of insulin resistance in the Asian-Indian men was associated with an approximately 2-fold increase in HTG content and plasma IL-6 concentrations compared with Caucasian men. These data demonstrate important ethnic and gender differences in the pathogenesis of insulin resistance in Asian-Indian men and have important therapeutic implications for treatment of T2DM and for the development of steatosis-related liver disease in this ethnic group. PMID- 17114291 TI - Elucidation of the function of type 1 human methionine aminopeptidase during cell cycle progression. AB - Processing of the N-terminal initiator methionine is an essential cellular process conserved from prokaryotes to eukaryotes. The enzymes that remove N terminal methionine are known as methionine aminopeptidases (MetAPs). Human MetAP2 has been shown to be required for the proliferation of endothelial cells and angiogenesis. The physiological function of MetAP1, however, has remained elusive. In this report we demonstrate that a family of inhibitors with a core structure of pyridine-2-carboxylic acid previously developed for the bacterial and yeast MetAP1 is also specific for human MetAP1 (HsMetAP1), as confirmed by both enzymatic assay and high-resolution x-ray crystallography. Treatment of tumor cell lines with the MetAP1-specific inhibitors led to an accumulation of cells in the G(2)/M phase, suggesting that HsMetAP1 may play an important role in G(2)/M phase transition. Overexpression of HsMetAP1, but not HsMetAP2, conferred resistance of cells to the inhibitors, and the inhibitors caused retention of N terminal methionine of a known MetAP substrate, suggesting that HsMetAP1 is the cellular target for the inhibitors. In addition, when HsMetAP1 was knocked down by gene-specific siRNA, cells exhibited slower progression during G(2)/M phase, a phenotype similar to cells treated with MetAP1 inhibitors. Importantly, MetAP1 inhibitors were able to induce apoptosis of leukemia cell lines, presumably as a consequence of their interference with the G(2)/M phase checkpoint. Together, these results suggest that MetAP1 plays an important role in G(2)/M phase of the cell cycle and that it may serve as a promising target for the discovery and development of new anticancer agents. PMID- 17114292 TI - Paclitaxel induces calcium oscillations via an inositol 1,4,5-trisphosphate receptor and neuronal calcium sensor 1-dependent mechanism. AB - Taxol (Paclitaxel) is an important natural product for the treatment of solid tumors. Despite a well documented tubulin-stabilizing effect, many side effects of taxol therapy cannot be explained by cytoskeletal mechanisms. In the present study submicromolar concentrations of taxol, mimicking concentrations found in patients, induced cytosolic calcium (Ca(2+)) oscillations in a human neuronal cell line. These oscillations were independent of extracellular and mitochondrial Ca(2+) but dependent on intact signaling via the phosphoinositide signaling pathway. We identified a taxol binding protein, neuronal Ca(2+) sensor 1 (NCS-1), a Ca(2+) binding protein that interacts with the inositol 1,4,5-trisphosphate receptor from a human brain cDNA phage display library. Taxol increased binding of NCS-1 to the inositol 1,4,5-trisphosphate receptor. Short hairpin RNA-mediated knockdown of NCS-1 in the same cell line abrogated the response to taxol but not to other agonists stimulating the phosphoinositide signaling pathway. These findings are important for studies involving taxol as a research tool in cell biology and may help to devise new strategies for the management of side effects induced by taxol therapy. PMID- 17114293 TI - NOTCH1 directly regulates c-MYC and activates a feed-forward-loop transcriptional network promoting leukemic cell growth. AB - The NOTCH1 signaling pathway directly links extracellular signals with transcriptional responses in the cell nucleus and plays a critical role during T cell development and in the pathogenesis over 50% of human T cell lymphoblastic leukemia (T-ALL) cases. However, little is known about the transcriptional programs activated by NOTCH1. Using an integrative systems biology approach we show that NOTCH1 controls a feed-forward-loop transcriptional network that promotes cell growth. Inhibition of NOTCH1 signaling in T-ALL cells led to a reduction in cell size and elicited a gene expression signature dominated by down regulated biosynthetic pathway genes. By integrating gene expression array and ChIP-on-chip data, we show that NOTCH1 directly activates multiple biosynthetic routes and induces c-MYC gene expression. Reverse engineering of regulatory networks from expression profiles showed that NOTCH1 and c-MYC govern two directly interconnected transcriptional programs containing common target genes that together regulate the growth of primary T-ALL cells. These results identify c-MYC as an essential mediator of NOTCH1 signaling and integrate NOTCH1 activation with oncogenic signaling pathways upstream of c-MYC. PMID- 17114294 TI - Participation of mouse DNA polymerase iota in strand-biased mutagenic bypass of UV photoproducts and suppression of skin cancer. AB - DNA polymerase iota (pol iota) is a conserved Y family enzyme that is implicated in translesion DNA synthesis (TLS) but whose cellular functions remain uncertain. To test the hypothesis that pol iota performs TLS in cells, we compared UV induced mutagenesis in primary fibroblasts derived from wild-type mice to mice lacking functional pol eta, pol iota, or both. A deficiency in mouse DNA polymerase eta (pol eta) enhanced UV-induced Hprt mutant frequencies. This enhanced UV-induced mutagenesis and UV-induced mutagenesis in wild-type cells were strongly diminished in cells deficient in pol iota, indicating that pol iota participates in the bypass of UV photoproducts in cells. Moreover, a clear strand bias among UV-induced base substitutions was observed in wild-type cells that was diminished in pol eta- and pol iota-deficient mouse cells and abolished in cells deficient in both enzymes. These data suggest that these enzymes bypass UV photoproducts in an asymmetric manner. To determine whether pol iota status affects cancer susceptibility, we compared the UV-induced skin cancer susceptibility of wild-type mice to mice lacking functional pol eta, pol iota, or both. Although pol iota deficiency alone had no effect, UV-induced skin tumors in pol eta-deficient mice developed 4 weeks earlier in mice concomitantly deficient in pol iota. Collectively, these data reveal functions for pol iota in bypassing UV photoproducts and in delaying the onset of UV-induced skin cancer. PMID- 17114295 TI - Mst3b, a purine-sensitive Ste20-like protein kinase, regulates axon outgrowth. AB - The growth of axons is fundamental to the development and repair of brain circuitry. We show here that Mst3b, a neuron-specific homolog of the yeast kinase Ste20, is critical for axon outgrowth. Mst3b is activated in response to trophic factors, and suppressing its expression (via siRNAs) or its function (by a dominant-negative mutant) blocks axon outgrowth. Inosine, a purine nucleoside that stimulates axon outgrowth, activates Mst3b kinase activity, whereas 6 thioguanine, a purine analog that blocks outgrowth, inhibits the activity of this kinase. These findings place Mst3b as a key regulator of axon outgrowth and help explain the purine sensitivity of this process. PMID- 17114296 TI - Prolyl hydroxylase-1 negatively regulates IkappaB kinase-beta, giving insight into hypoxia-induced NFkappaB activity. AB - Hypoxia is a feature of the microenvironment of a growing tumor. The transcription factor NFkappaB is activated in hypoxia, an event that has significant implications for tumor progression. Here, we demonstrate that hypoxia activates NFkappaB through a pathway involving activation of IkappaB kinase-beta (IKKbeta) leading to phosphorylation-dependent degradation of IkappaBalpha and liberation of NFkappaB. Furthermore, through increasing the pool and/or activation potential of IKKbeta, hypoxia amplifies cellular sensitivity to stimulation with TNFalpha. Within its activation loop, IKKbeta contains an evolutionarily conserved LxxLAP consensus motif for hydroxylation by prolyl hydroxylases (PHDs). Mimicking hypoxia by treatment of cells with siRNA against PHD-1 or PHD-2 or the pan-prolyl hydroxylase inhibitor DMOG results in NFkappaB activation. Conversely, overexpression of PHD-1 decreases cytokine-stimulated NFkappaB reporter activity, further suggesting a repressive role for PHD-1 in controlling the activity of NFkappaB. Hypoxia increases both the expression and activity of IKKbeta, and site-directed mutagenesis of the proline residue (P191A) of the putative IKKbeta hydroxylation site results in a loss of hypoxic inducibility. Thus, we hypothesize that hypoxia releases repression of NFkappaB activity through decreased PHD-dependent hydroxylation of IKKbeta, an event that may contribute to tumor development and progression through amplification of tumorigenic signaling pathways. PMID- 17114297 TI - Structural adaptations in the murine colon microcirculation associated with hapten-induced inflammation. AB - BACKGROUND: Blood flowing across the vascular endothelium creates wall shear stress, dependent on velocity of flow and vessel geometry, that tends to disrupt lymphocyte-endothelial cell adhesion. OBJECTIVE: The microcirculation in a murine model of acute colitis was investigated to identify structural adaptations during acute colitis that may facilitate transmigration. METHODS: In 2,4,6 trinitrobenzenesulphonic acid-induced acute colitis, the infiltrating cells and colonic microcirculation was investigated by cellular topographic mapping, corrosion casting and three-dimensional scanning electron microscopy (SEM). Colonic blood velocimetry was performed using intravital microscopy. RESULTS: Clinical and histological parameters suggested a peak inflammatory response at 96 h (p<0.001). The infiltrating cells were spatially related to the mucosal capillary plexus by three-dimensional topographic mapping (p<0.001). In normal mice, corrosion casting and three-dimensional SEM showed a polygonal mucosal plexus supplied by ascending arteries and descending veins. After 2,4,6 trinitrobenzenesulphonic acid stimulation, three-dimensional SEM showed preserved branch angles (p = 0.52) and nominal vessel lengths (p = 0.93), but a significantly dilated mucosal capillary plexus (p<0.001). Intravital microscopy of the mucosal plexus showed a greater than twofold decrease in the velocity of flow (p<0.001). CONCLUSIONS: The demonstrable slowing of the velocity of flow despite an increase in volumetric flow suggests that these microvascular adaptations create conditions suitable for leucocyte adhesion and transmigration. PMID- 17114298 TI - Protease-activated receptor-2 protects against pancreatitis by stimulating exocrine secretion. AB - BACKGROUND: Protease-activated receptor-2 (PAR-2) is present in the pancreas, where it has been shown to play a protective role during pancreatitis. However, the mechanism by which it protects against pancreatitis still remains to be elucidated. Acute pancreatitis is associated with premature zymogen activation and a blockage in digestive enzyme secretion. AIM: To examine the effects of PAR 2 activation on the severity of pancreatitis, and to determine whether its protective effects are mediated by affecting either premature activation or secretory blockage, or both. RESULTS: The results confirmed that PAR-2 -/- mice have more severe pancreatitis than wild-type mice. Interestingly, intrapancreatic trypsin levels in the PAR-2 knockouts remained high after 6 h of pancreatitis, whereas they reverted to normal in the wild types. During pancreatitis, PAR-2 mRNA levels were upregulated in wild-type mice in response to supramaximal caerulein administration. Further, after a single injection of supramaximal caerulein, PAR-2 mRNA levels were also elevated, reaching a peak at 3 h. Stimulating PAR-2 with trypsin or the PAR-2-activating peptide, serine-leucine isoleucine-glycine-arginine-leucine (SLIGRL), induced significantly more secretion from the acini of these caerulein-sensitised mice compared with the controls. PAR-2 activation also reversed the inhibition of secretion observed in both the caerulein and arginine models. CONCLUSIONS: Trypsin released during the early stages of pancreatitis activates PAR-2 receptors on the acinar cells and stimulates secretion from these cells. Thus, PAR-2 activation may decrease pancreatic injury and limit the severity of pancreatitis by allowing extracellular trypsin to act as a secretagogue. PMID- 17114299 TI - Benzo[a]pyrene-7,8-dihydrodiol promotes checkpoint activation and G2/M arrest in human bronchoalveolar carcinoma H358 cells. AB - Polycyclic aromatic hydrocarbons (PAHs) are potent carcinogens that require metabolic activation inside cells. The proximate carcinogens PAH-diols can be converted to o-quinones by aldo-keto reductases (AKRs) or to diol-epoxides by cytochrome P450 (P450) enzymes. We assessed the effect of benzo[a]pyrene-7,8 dihydrodiol (BPD) on proliferation in p53-null bronchoalveolar carcinoma H358 cells. BPD treatment led to a significant inhibition of proliferation and arrest in G2/M in H358 cells. The relative contribution of the AKR and P450 pathways to cell cycle arrest was assessed. Overexpression of AKR1A1 did not affect cell proliferation or cell cycle progression, and benzo[a]pyrene-7,8-dione did not cause any noticeable effect on cell growth, suggesting that AKR1A1 metabolic products were not involved in the antiproliferative effect of BPD. On the other hand, blockade of P450 induction or inhibition of P450 activity greatly impaired the effect of BPD. Moreover, P450 induction by 2,3,7,8-tetrachlorodibenzo-p dioxin significantly enhanced the antiproliferative effect of BPD. Mechanistic studies revealed that BPD caused a DNA damage response, Chk1 activation, and accumulation of phospho-Cdc2 (Tyr15) in H358 cells, effects that were impaired by an ataxia-telangectasia mutated (ATM)/ATM-related (ATR) inhibitor. Similar results were observed in human bronchoepithelial BEAS-2B cells, arguing for analogous mechanisms in tumorigenic and immortalized nontumorigenic cells lacking functional p53. Our data suggest that a p53-independent pathway operates in lung epithelial cells in response to BPD that involves P450 induction and subsequent activation of the ATR/ATM/Chk1 damage check-point pathway and cell cycle arrest in G2/M. PMID- 17114300 TI - Satisfaction with present life predicts survival in octogenarians. AB - We examined the effect of life satisfaction on survival over 10 years among 80 year-old and older same-sex twins of whom 320 individuals responded to the Life Satisfaction Index Z questionnaire in connection with the OCTO-Twin study. We treated participants as individuals in semiparametric Cox regression mixed effects models (frailty) by adjusting the similarity of mortality risk within twin pairs by modeling it as a random variable. An exploratory factor analysis yielded three factors: Zest and Mood represented satisfaction with present life and Congruence represented satisfaction with past life. Those in the lowest quartile of factors of satisfaction with present life had an almost twofold risk for mortality compared with those in the highest quartile, even after adjustment for multiple confounders. Satisfaction with past life satisfaction showed no association with mortality. PMID- 17114301 TI - Age differences in memory-load interference effects in syntactic processing. AB - The effects of a memory load on syntactic processing by younger and older adults were examined. Participants were asked to remember a noun phrase (NP) memory load while they read sentences varying in syntactic complexity. Two types of NPs were used as memory loads: proper names or definite descriptions referring to occupations or roles. The NPs used in the sentence and memory load either matched (e.g., all proper names or all occupations), or they mismatched. Participants read complex sentences more slowly than they did simpler sentences; for young adults, this complexity effect was exacerbated when memory interference was generated by matching NPs in the sentence and memory load, whereas for older adults, memory-load interference did not vary with sentence complexity or memory load matching. These results suggest that a general reduction in older adults' processing capacity was produced by the memory load, whereas the matching memory loads and sentence NPs produced a more specific form of interference that affected young adults' online processing. PMID- 17114302 TI - Perceptions of forgetful and slow employees: does age matter? AB - Participants (perceivers) read a vignette describing a young or older employee (target) in a young-relevant or old-relevant work context who is either forgetful or slow. Regardless of work context, perceivers attributed older targets' forgetful and slow behavior more to internal stable causes but the identical behavior of younger targets more to internal unstable causes. Perceivers also felt less anger and greater sympathy and were more likely to recommend a promotion and raise for older than for young targets. Perceivers' anger for young and older targets was mediated by their internal unstable attributions, but their sympathy was not mediated by their internal stable attributions. Perceivers' promotion and raise recommendations were associated with the degree of sympathy they felt more for young than for older targets. PMID- 17114303 TI - Release from implicit interference in memory and metamemory: older adults know that they can't let go. AB - Cued recall performance is better when cue and targets have a small number of semantic associates, which is an effect of implicit interference from shared associates ( Nelson, McKinney, Gee, & Janczura, 1998). The present study examined age-related effects on memory under conditions of implicit interference. Recall and recognition performance of both younger and older adults was evaluated for small- versus large-set-size cues under two contexts. Comparable cue-set-size effects were obtained for both age groups under extralist cueing, but they were eliminated only for younger adults under intralist cueing. Older adults were not able to use the context to effectively eliminate implicit interference from associates of the cue as did younger adults, perhaps because of an inhibition deficit. Both groups had equivalent metamemory accuracy and sensitivity, indicating that the monitoring of learning prior to a test reflected the effects of implicit interference and is not impaired by aging. PMID- 17114304 TI - Effects of repeated testing in a longitudinal age-homogeneous study of cognitive aging. AB - Estimates of gains related to repeated test exposure (retest effects) and within person cognitive changes are confounded in most longitudinal studies because of the nonindependent time structures underlying both processes. Recently developed statistical approaches rely on between-person age differences to estimate effects of repeated testing. This study, however, demonstrates how retest effects can be evaluated at the group level in an age-homogeneous population-based study by use of a sampling-based design approach in which level and change of cognitive performance of previous participants, measured at ages 70, 75, 79, 81, 85, 88, 90, 92, 95, 97, and 99 years, were compared with performances of survivors of a representative sample identified and drawn from the same original population cohort but invited for the first time at age 85 with subsequent measurements at ages 88, 90, 92, 95, 97, and 99. The comparisons revealed a trend toward retest effects on two out of five cognitive measurements. The study demonstrates how a design-based approach can provide valuable insights into continuous learning processes embedded in population average aging trajectories that are not confounded with cohort and mortality-related selective attrition. PMID- 17114305 TI - Twin study of depressive symptoms among older african-american women. AB - This study examines factors associated with depressive symptoms in a genetically informative sample of African-American female twins aged 65 years and older. A telephone interview was conducted with 180 pairs of twins. Questions included demographics, health behaviors, health status, activities of daily living (ADLs), instrumental ADLs, and depressive symptoms as measured by the Center for Epidemiologic Studies-Depression scale. Regression methods for clustered data were used to examine the associations. In univariate analyses, ADLs (odds ratio or OR = 1.4, 95% confidence interval or CI = 1.1-1.7), fractures (OR = 4.4, 95% CI = 1.3-15.6), and vision problems (OR = 1.9, 95% CI = 1.0-3.8) were significantly associated with depressive symptoms. In multivariable analyses, ADLs (OR = 1.4, 95% CI = 1.2-1.7) and vision problems (OR = 2.0, 95% CI = 1.2 3.5) remained significantly associated with depressive symptoms. A within-pair analysis, controlling for genetic or familial influences, produced similar results. The results suggest that efforts targeted at reducing levels of disability may reduce depressive symptoms in this population. PMID- 17114306 TI - Personality and perceived health in older adults: the five factor model in primary care. AB - Responses to specific questions tapping perceived health are associated with morbidity, mortality, and the use of health services, yet there has been little research on their personality correlates. We examined the associations between Five Factor Model personality traits and responses to four items extracted from the Medical Outcomes Study Short Form-36 in 266 primary care patients who were 65 years of age or older. Multivariate analyses controlling for age, gender, depressive symptoms, and physical disease burden showed that having a higher Neuroticism score was associated with worse perceived health in response to all items except "I am as healthy as anybody I know." Having a lower Extraversion score was associated with worse perceived health in response to the item "I expect my health to get worse." We discuss implications for understanding personality influences on morbidity, mortality, and health services utilization. PMID- 17114307 TI - Saying versus touching: age differences in short-term memory are affected by the type of response. AB - We examined whether the type of response used to report items recalled from short term memory affects the age difference in verbal and spatial memory spans. Younger and older adults viewed either a series of letters or a series of locations in a grid, and then they reported their memory for the items either vocally or by using a touch screen. Overall, age differences were larger for spatial memory spans than for verbal memory spans, replicating previous results. Changing the response modality affected only older adults' verbal spans, which were approximately one item higher with a vocal response than with a manual response. This resulted in a smaller age difference for verbal items reported vocally than for any other condition. The results can best be explained by age related difficulties in both spatial processing and in dealing with stimulus response incongruity. PMID- 17114308 TI - Social status, risky health behaviors, and diabetes in middle-aged and older adults. AB - OBJECTIVE: This article investigates: (a) how social status influences diabetes prevalence and incidence; (b) how risky health behaviors contribute to the prediction of incident diabetes; (c) if the effects of health behaviors mediate the effects of social status on incident diabetes; and (d) if these effects differ in midlife and older age. METHODS: We examined nationally representative data from the 1992/1993-1998 panels of the Health and Retirement Study for middle aged and older adults using logistic regression analyses. RESULT: The odds of prevalent diabetes were higher for people of older age, men, Black adults, and Latino adults. Higher early-life social status (e.g., parental schooling) and achieved social status (e.g., respondent schooling, economic resources) reduced the odds in both age groups. We observed similar patterns for incident diabetes in midlife but not in older age. Risky health behaviors--particularly obesity- increased the odds of incident diabetes in both age groups independent of social status. The increased odds of incident diabetes in midlife persisted for Black and Latino adults net of other social status factors. DISCUSSION: Risky health behaviors are key predictors of incident diabetes in both age groups. Economic resources also play an important protective role in incident diabetes in midlife but not in older age. PMID- 17114309 TI - The color of hospitalization over the adult life course: cumulative disadvantage in black and white? AB - OBJECTIVES: Drawing from cumulative disadvantage theory, this research addresses the following questions: Do hospital admission and discharge rates differ for White and Black adults? If yes, do the differences amplify in later life? METHODS: This study made use of hospital records abstracted from a long-term prospective study of adults in the National Health and Nutrition Examination Survey I: Epidemiologic Follow-up Study (N = 6,833). Semi-Markov models were specified to examine the likelihood of hospital admission and discharge for Black and White adults aged 25 to 74 years old at baseline. RESULTS: Black adults were less likely than White adults to be admitted to the hospital, but they had longer lengths of stay. The risk of death in the hospital was greater for both Black men and women than for White men and women. In addition, the observed racial differences in hospitalization experiences amplified in later life. DISCUSSION: Health inequality in America is manifest in how White and Black adults enter and exit hospitals. The findings demonstrate growing heterogeneity in later life by race. PMID- 17114310 TI - The impact of comorbidity on wealth changes in later life. AB - OBJECTIVES: Despite the high prevalence of comorbidity in later life, scientists do not fully understand its financial impact. The objective of this study was to enhance researchers' understanding of the impact of compounded health problems on the wealth of older people. METHODS: Using data from the Asset and Health Dynamics Among the Oldest Old study (1995 to 2002 waves), we conducted ordinary least squares regression analysis on wealth changes. RESULTS: We found that comorbidity leads to significant wealth depletion in later life, especially for single elders. Single elders with comorbidity depleted 20% to 22% of their wealth over a 2- to 3-year time period, especially those with the combination of heart disease and diabetes. The impact of comorbidity was disproportionately greater than the estimated impact of a single health problem. However, the impact of comorbidity did not appear to be significant among married people. DISCUSSION: We found that compounded health problems also create compounded financial problems in later life. For an accurate estimation of the financial consequences of health problems, it is important to consider comorbid health problems, as the effect of comorbidity is not equal to the sum of the effects of single health problems. PMID- 17114311 TI - Strains of singlehood in later life: do race and gender matter? AB - OBJECTIVES: Few studies have identified the distinctive aspects of singlehood that are distressing to older adults. The objectives of our study were: (a) to examine whether divorced, widowed, and never-married older adults differed in their experiences of single strain, an indicator of chronic stressors associated with being unmarried; and (b) to assess whether the marital status differences we explored varied by gender and race. METHODS: Using data from a subsample of 530 unmarried older adults and ordinary least squares regression, we estimated main and interactive effects of marital status, gender, and race on single strain. RESULTS: Divorced and widowed persons reported higher single strain than never married persons, although the magnitude of these effects varied considerably by race and gender. Never-married White women reported higher levels of single strain than their male counterparts. White widows and widowers exhibited higher single strain than widowed Black adults. Black women uniformly fared better than White women, whereas divorced and never-married Black men were not different from their White peers in terms of single strain. DISCUSSION: Psychological adjustment to singlehood among older adults reflects patterns of gender and race stratification and socialization over the life course. PMID- 17114312 TI - Attrition of older adults in longitudinal surveys: detection and correction of sample selection bias using multigenerational data. AB - OBJECTIVE: The purpose of this study was (a) to investigate whether attrition due to death and nonresponse leads to bias in estimated growth-decline trajectories when only complete data are used in longitudinal research, and (b) to examine the extent of the bias and possible solutions. METHODS: The study sample was a subset of the Longitudinal Study of Generations and included data from 208 G1-G2 parent child dyads and 538 G2-G3 dyads over 30 years. We used a latent growth-decline curve model based on full information maximum likelihood estimation in order to compare parents' and adult children's reports on older respondents' health and intergenerational solidarity by parents' attrition status. RESULT: Results indicated that attrition due to mortality biased estimates of respondents' assessments of their functional health status over time, and parents' perceptions of the quality of the parent-child relationship deteriorated more rapidly among those who died by Time 7, but nonresponse did not seriously bias estimates of these measures. Using proxies, we found that functional impairment increased more rapidly when children reported about parents, especially in advanced old age. DISCUSSION: These results support the use of full information in estimating growth curves where mortality is present but raise concerns when using child proxies to evaluate parental health or the quality of intergenerational relationships. PMID- 17114313 TI - A mixed-methods approach to understanding loneliness and depression in older adults. AB - OBJECTIVE: Depression in late life may be difficult to identify, and older adults often do not accept depression treatment offered. This article describes the methods by which we combined an investigator-defined definition of depression with a person-derived definition of depression in order to understand how older adults and their primary care providers overlapped and diverged in their ideas about depression. METHODS: We recruited a purposive sample of 102 persons aged 65 years and older with and without significant depressive symptoms on a standardized assessment scale (Center for Epidemiologic Studies-Depression scale) from primary care practices and interviewed them in their homes. We applied methods derived from anthropology and epidemiology (consensus analysis, semi structured interviews, and standardized assessments) in order to understand the experience and expression of late-life depression. RESULT: Loneliness was highly salient to older adults whom we asked to describe a depressed person or themselves when depressed. Older adults viewed loneliness as a precursor to depression, as self-imposed withdrawal, or as an expectation of aging. In structured interviews, loneliness in the week prior to interview was highly associated with depressive symptoms, anxiety, and hopelessness. DISCUSSION: An improved understanding of how older adults view loneliness in relation to depression, derived from multiple methods, may inform clinical practice. PMID- 17114314 TI - Continuity of leisure participation from middle age to old age. AB - OBJECTIVE: Continuity in habits, activities, and roles is important upon entering old age according to the continuity theory of aging. Few studies have investigated patterns of leisure participation over an extended period of time among older adults. This study examines changes in nine different leisure activities in a nationally representative sample of individuals followed over a 34-year period in Sweden. METHODS: We used longitudinal data from three waves of an interview survey that followed 495 individuals from 1968 to 2002. Individuals were aged 43-65 in 1968 and 77-99 in 2002. We conducted logistic regression analyses on each of the leisure activities. RESULT: For the panel followed, a decline in participation rates was the most common pattern over time. Analyses at the individual level showed that late-life participation was generally preceded by participation earlier in life. Previous participation, both 10 and 34 years earlier, predicted late-life participation. The modifying effect of functional status in late life was small. DISCUSSION: In accordance with the continuity theory of aging, leisure participation in old age is often a continuation of previous participation. However, there is considerable variation among both activities and individuals. PMID- 17114315 TI - Driving cessation and consumption expenses in the later years. AB - OBJECTIVES: This study examined the association between consumption and driving status among older persons within the context of selected variables, including self-rated health and functional status. METHODS: The data were from the 1998, 2000, and 2002 Health and Retirement Study and the 2003 Health and Retirement Study Consumption and Activities Mail Survey. We conducted Tobit regression analyses on five consumption categories of basic needs (such as food) and higher order needs (such as trips and dining out). RESULTS: Consumption and driving status were significantly associated, showing that driving cessation was related to a 46% to 63% reduction in spending on trips, tickets, and dining out. Another significant relationship emerged between consumption and having never driven. Driving cessation was minimally related to consumption of basic needs (such as food and clothing) and was more strongly associated with higher order needs (such as trips). DISCUSSION: The findings demonstrate the association between older people's driving status and consumption, specifically higher order activities. Older persons who drive and, presumably, have more opportunities to go to stores, restaurants, and other outside events, spend more on food, tickets, and dining out than those who cease driving or have never driven. Although the direction of causality remains unclear, these findings have implications for those concerned with alternative transportation resources for older adults. PMID- 17114316 TI - Novel role for Aeromonas jandaei as a digestive tract symbiont of the North American medicinal leech. AB - The gut bacteria of the North American medicinal leech, Macrobdella decora, were characterized. Biochemical tests and DNA sequences indicated that Aeromonas jandaei is the dominant culturable symbiont in leeches from a broad geographic area. In this work we identified a new habitat for A. jandaei, and here we suggest that there is unexpected specificity between leeches and Aeromonas species. PMID- 17114317 TI - Degradation and mineralization of nanomolar concentrations of the herbicide dichlobenil and its persistent metabolite 2,6-dichlorobenzamide by Aminobacter spp. isolated from dichlobenil-treated soils. AB - 2,6-Dichlorobenzamide (BAM), a persistent metabolite from the herbicide 2,6 dichlorobenzonitrile (dichlobenil), is the pesticide residue most frequently detected in Danish groundwater. A BAM-mineralizing bacterial community was enriched from dichlobenil-treated soil sampled from the courtyard of a former plant nursery. A BAM-mineralizing bacterium (designated strain MSH1) was cultivated and identified by 16S rRNA gene sequencing and fatty acid analysis as being closely related to members of the genus Aminobacter, including the only cultured BAM degrader, Aminobacter sp. strain ASI1. Strain MSH1 mineralized 15 to 64% of the added [ring-U-(14)C]BAM to (14)CO(2) with BAM at initial concentrations in the range of 7.9 nM to 263.1 muM provided as the sole carbon, nitrogen, and energy source. A quantitative enzyme-linked immunoassay analysis with antibodies against BAM revealed residue concentrations of 0.35 to 18.05 nM BAM following incubation for 10 days, corresponding to a BAM depletion of 95.6 to 99.9%. In contrast to the Aminobacter sp. strain ASI1, strain MSH1 also mineralized the herbicide itself along with several metabolites, including ortho chlorobenzonitrile, ortho-chlorobenzoic acid, and benzonitrile, making it the first known dichlobenil-mineralizing bacterium. Aminobacter type strains not previously exposed to dichlobenil or BAM were capable of degrading nonchlorinated structural analogs. Combined, these results suggest that closely related Aminobacter strains may have a selective advantage in BAM-contaminated environments, since they are able to use this metabolite or structurally related compounds as a carbon and nitrogen source. PMID- 17114318 TI - Phylogenetic analysis of the pPT23A plasmid family of Pseudomonas syringae. AB - The pPT23A plasmid family of Pseudomonas syringae contains members that contribute to the ecological and pathogenic fitness of their P. syringae hosts. In an effort to understand the evolution of these plasmids and their hosts, we undertook a comparative analysis of the phylogeny of plasmid genes and that of conserved chromosomal genes from P. syringae. In total, comparative sequence and phylogenetic analyses were done utilizing 47 pPT23A family plasmids (PFPs) from 16 pathovars belonging to six genomospecies. Our results showed that the plasmid replication gene (repA), the only gene currently known to be distributed among all the PFPs, had a phylogeny that was distinct from that of the P. syringae hosts of these plasmids and from those of other individual genes on PFPs. The phylogenies of two housekeeping chromosomal genes, those for DNA gyrase B subunit (gyrB) and primary sigma factor (rpoD), however, were strongly associated with genomospecies of P. syringae. Based on the results from this study, we conclude that the pPT23A plasmid family represents a dynamic genome that is mobile among P. syringae pathovars. PMID- 17114319 TI - Functional analysis of Burkholderia cepacia genes bceD and bceF, encoding a phosphotyrosine phosphatase and a tyrosine autokinase, respectively: role in exopolysaccharide biosynthesis and biofilm formation. AB - The biosynthesis of the exopolysaccharide (EPS) cepacian by Burkholderia cepacia complex strains requires the 16.2-kb bce cluster of genes. Two of the clustered genes, bceD and bceF, code for two proteins homologous to phosphotyrosine phosphatases and tyrosine kinases, respectively. We show experimental evidence indicating that BceF is phosphorylated on tyrosine and that the conserved lysine residue present at position 563 in the Walker A ATP-binding motif is required for this autophosphorylation. It was also proved that BceD is capable of dephosphorylating the phosphorylated BceF. Using the artificial substrate p nitrophenyl phosphate (PNPP), BceD exhibited a V(max) of 8.8 mumol of PNPP min( 1) mg(-1) and a K(m) of 3.7 mM PNPP at 30 degrees C. The disruption of bceF resulted in the abolishment of cepacian accumulation in the culture medium, but 75% of the parental strain's EPS production yield was still registered for the bceD mutant. The exopolysaccharide produced by the bceD mutant led to less viscous solutions and exhibited the same degree of acetylation as the wild-type cepacian, suggesting a lower molecular mass for this mutant biopolymer. The size of the biofilm produced in vitro by bceD and bceF mutant strains is smaller than the size of the biofilm formed by the parental strain, and this phenotype was confirmed by complementation assays, indicating that BceD and BceF play a role in the establishment of biofilms of maximal size. PMID- 17114320 TI - Genes that enhance the ecological fitness of Shewanella oneidensis MR-1 in sediments reveal the value of antibiotic resistance. AB - Environmental bacteria persist in various habitats, yet little is known about the genes that contribute to growth and survival in their respective ecological niches. Signature-tagged mutagenesis (STM) of Shewanella oneidensis MR-1 coupled with a screen involving incubations of mutant strains in anoxic aquifer sediments allowed us to identify 47 genes that enhance fitness in sediments. Gene functions inferred from annotations provide us with insight into physiological and ecological processes that environmental bacteria use while growing in sediment ecosystems. Identification of the mexF gene and other potential membrane efflux components by STM demonstrated that homologues of multidrug resistance genes present in pathogens are required for sediment fitness of nonpathogenic bacteria. Further studies with a mexF deletion mutant demonstrated that the multidrug resistance pump encoded by mexF is required for resistance to antibiotics, including chloramphenicol and tetracycline. Chloramphenicol-adapted cultures exhibited mutations in the gene encoding a TetR family regulatory protein, indicating a role for this protein in regulating expression of the mexEF operon. The relative importance of mexF for sediment fitness suggests that antibiotic efflux may be a required process for bacteria living in sediment systems. PMID- 17114321 TI - Use of quantum dot luminescent probes to achieve single-cell resolution of human oral bacteria in biofilms. AB - Oral biofilms are multispecies communities, and in their nascent stages of development, numerous bacterial species engage in interspecies interactions. Better insight into the spatial relationship between different species and how species diversity increases over time can guide our understanding of the role of interspecies interactions in the development of the biofilms. Quantum dots (QD) are semiconductor nanocrystals and have emerged as a promising tool for labeling and detection of bacteria. We sought to apply QD-based primary immunofluorescence for labeling of bacterial cells with in vitro and in vivo biofilms and to compare this approach with the fluorophore-based primary immunofluorescence approach we have used previously. To investigate QD-based primary immunofluorescence as the means to detect distinct targets with single-cell resolution, we conjugated polyclonal and monoclonal antibodies to the QD surface. We also conducted simultaneous QD conjugate-based and fluorophore conjugate-based immunofluorescence and showed that these conjugates were complementary tools in immunofluorescence applications. Planktonic and biofilm cells were labeled effectively by considering two factors: the final nanomolar concentration of QD conjugate and the amount of antibody conjugated to the QD, which we define as the degree of labeling. These advances in the application of QD-based immunofluorescence for the study of biofilms in vitro and in vivo will help to define bacterial community architecture and to facilitate investigations of interactions between bacterial species in these communities. PMID- 17114322 TI - Influence of dangling ends and surface-proximal tails of targets on probe-target duplex formation in 16S rRNA gene-based diagnostic arrays. AB - Dangling ends and surface-proximal tails of gene targets influence probe-target duplex formation and affect the signal intensity of probes on diagnostic microarrays. This phenomenon was evaluated using an oligonucleotide microarray containing 18-mer probes corresponding to the 16S rRNA genes of 10 waterborne pathogens and a number of synthetic and PCR-amplified gene targets. Signal intensities for Klenow/random primer-labeled 16S rRNA gene targets were dissimilar from those for 45-mer synthetic targets for nearly 73% of the probes tested. Klenow/random primer-labeled targets resulted in an interaction with a complex mixture of 16S rRNA genes (used as the background) 3.7 times higher than the interaction of 45-mer targets with the same mixture. A 7-base-long dangling end sequence with perfect homology to another single-stranded background DNA sequence was sufficient to produce a cross-hybridization signal that was as strong as the signal obtained by the probe-target duplex itself. Gibbs free energy between the target and a well-defined background was found to be a better indicator of hybridization signal intensity than the sequence or length of the dangling end alone. The dangling end (Gibbs free energy of -7.6 kcal/mol) was found to be significantly more prone to target-background interaction than the surface-proximal tail (Gibbs free energy of -64.5 kcal/mol). This study underlines the need for careful target preparation and evaluation of signal intensities for diagnostic arrays using 16S rRNA and other gene targets due to the potential for target interaction with a complex background. PMID- 17114323 TI - Role of branched-chain fatty acids in pH stress tolerance in Listeria monocytogenes. AB - In alkaline conditions, Listeria monocytogenes cells develop higher proportions of branched-chain fatty acids (FAs), including more anteiso forms. In acid conditions, the opposite occurs. Reduced growth of pH-sensitive mutants at adverse pH (5.0/9.0) was alleviated by the addition of 2-methylbutyrate (an anteiso-FA precursor), suggesting that anteiso-FAs are important in adaptation to adverse pH. The balance between anteiso- and iso-FAs may be more important than changes in the amounts and/or degrees of saturation of FAs in pH adaptation. PMID- 17114324 TI - Sulfur-oxidizing bacteria in Soap Lake (Washington State), a meromictic, haloalkaline lake with an unprecedented high sulfide content. AB - Culture-dependent and -independent techniques were used to study the diversity of chemolithoautotrophic sulfur-oxidizing bacteria in Soap Lake (Washington State), a meromictic, haloalkaline lake containing an unprecedentedly high sulfide concentration in the anoxic monimolimnion. Both approaches revealed the dominance of bacteria belonging to the genus Thioalkalimicrobium, which are common inhabitants of soda lakes. A dense population of Thioalkalimicrobium (up to 10(7) cells/ml) was found at the chemocline, which is characterized by a steep oxygen sulfide gradient. Twelve Thioalkalimicrobium strains exhibiting three different phenotypes were isolated in pure culture from various locations in Soap Lake. The isolates fell into two groups according to 16S rRNA gene sequence analysis. One of the groups was closely related to T. cyclicum, which was isolated from Mono Lake (California), a transiently meromictic, haloalkaline lake. The second group, consisting of four isolates, was phylogenetically and phenotypically distinct from known Thioalkalimicrobium species and unique to Soap Lake. It represented a new species, for which we suggest the name Thioalkalimicrobium microaerophilum sp. nov. PMID- 17114325 TI - Biotransformation of patulin by Gluconobacter oxydans. AB - A bacterium isolated from patulin-contaminated apples was capable of degrading patulin to a less-toxic compound, ascladiol. The bacterium was identified as Gluconobacter oxydans by 16S rRNA gene sequencing, whereas ascladiol was identified by liquid chromatography-tandem mass spectrometry and proton and carbon nuclear magnetic resonance. Degradation of up to 96% of patulin was observed in apple juices containing up to 800 microg/ml of patulin and incubated with G. oxydans. PMID- 17114326 TI - Molecular analysis of arsenate-reducing bacteria within Cambodian sediments following amendment with acetate. AB - The health of millions is threatened by the use of groundwater contaminated with sediment-derived arsenic for drinking water and irrigation purposes in Southeast Asia. The microbial reduction of sorbed As(V) to the potentially more mobile As(III) has been implicated in release of arsenic into groundwater, but to date there have been few studies of the microorganisms that can mediate this transformation in aquifers. With the use of stable isotope probing of nucleic acids, we present evidence that the introduction of a proxy for organic matter ((13)C-labeled acetate) stimulated As(V) reduction in sediments collected from a Cambodian aquifer that hosts arsenic-rich groundwater. This was accompanied by an increase in the proportion of prokaryotes closely related to the dissimilatory As(V)-reducing bacteria Sulfurospirillum strain NP-4 and Desulfotomaculum auripigmentum. As(V) respiratory reductase genes (arrA) closely associated with those found in Sulfurospirillum barnesii and Geobacter uraniumreducens were also detected in active bacterial communities utilizing (13)C-labeled acetate in microcosms. This study suggests a direct link between inputs of organic matter and the increased prevalence and activity of organisms which transform As(V) to the potentially more mobile and thus hazardous As(III) via dissimilatory As(V) reduction. PMID- 17114327 TI - Use of Tetrahymena thermophila to study the role of protozoa in inactivation of viruses in water. AB - The ability of a ciliate to inactivate bacteriophage was studied because these viruses are known to influence the size and diversity of bacterial populations, which affect nutrient cycling in natural waters and effluent quality in sewage treatment, and because ciliates are ubiquitous in aquatic environments, including sewage treatment plants. Tetrahymena thermophila was used as a representative ciliate; T4 was used as a model bacteriophage. The T4 titer was monitored on Escherichia coli B in a double-agar overlay assay. T4 and the ciliate were incubated together under different conditions and for various times, after which the mixture was centrifuged through a step gradient, producing a top layer free of ciliates. The T4 titer in this layer decreased as coincubation time increased, but no decrease was seen if phage were incubated with formalin-fixed Tetrahymena. The T4 titer associated with the pellet of living ciliates was very low, suggesting that removal of the phage by Tetrahymena inactivated T4. When Tetrahymena cells were incubated with SYBR gold-labeled phage, fluorescence was localized in structures that had the shape and position of food vacuoles. Incubation of the phage and ciliate with cytochalasin B or at 4 degrees C impaired T4 inactivation. These results suggest the active removal of T4 bacteriophage from fluid by macropinocytosis, followed by digestion in food vacuoles. Such ciliate virophagy may be a mechanism occurring in natural waters and sewage treatment, and the methods described here could be used to study the factors influencing inactivation and possibly water quality. PMID- 17114328 TI - Role of zearalenone lactonase in protection of Gliocladium roseum from fungitoxic effects of the mycotoxin zearalenone. AB - Zearalenone is a mycotoxin with estrogenic effects on mammals that is produced by several species of Fusarium. We found that zearalenone and its derivatives inhibit the growth of filamentous fungi on solid media at concentrations of < or =10 microg/ml. The fungitoxic effect declined in the order zearalenone > alpha zearalenol > beta-zearalenol. The mycoparasitic fungus Gliocladium roseum produces a zearalenone-specific lactonase which catalyzes the hydrolysis of zearalenone, followed by a spontaneous decarboxylation. The growth of G. roseum was not inhibited by zearalenone, and the lactonase may protect G. roseum from the toxic effects of this mycotoxin. We inactivated zes2, the gene encoding zearalenone lactonase in G. roseum, by inserting a hygromycin resistance cassette into the coding sequence of the gene by means of Agrobacterium tumefaciens mediated genetic transformation. The zes2 disruption mutants could not hydrolyze the lactone bond of zearalenone and were more sensitive to zearalenone. These data are consistent with a hypothesis that resorcylic acid lactones exemplified by zearalenone act to reduce growth competition by preventing competing fungi from colonizing substrates occupied by zearalenone producers and suggest that they may play a role in fungal defense against mycoparasites. PMID- 17114329 TI - Bioluminescence resonance energy transfer as a screening assay: Focus on partial and inverse agonism. AB - The reported data for compound screening with the bioluminescence resonance energy transfer (BRET2) assay is very limited, and several questions remain unaddressed, such as the behavior of agonists. Eleven beta2 adrenergic receptor (beta2-AR) agonists were tested for full or partial agonism in an improved version of the receptor/beta-arrestin2 BRET2 assay and in 2 cyclic adenosine monophosphate (cAMP) assays (column cAMP assay and ALPHAscreen cAMP assay). Tested in the highly sensitive ALPHAscreen cAMP assay, all selected agonists behaved as full agonists, using isoproterenol as a reference compound. In the less sensitive column cAMP assay, ephedrine and dopamine had a clear partial response. For the BRET2 assay, a highly graded picture was obtained. Moreover, beta2-AR antagonists were tested for inverse agonism. Pronounced inverse agonism was detected in the ALPHAscreen cAMP assay. Only marginal inverse agonistic responses were seen for alprenolol and pindolol in the column cAMP assay, and no inverse agonism was seen in the BRET2 assay. For the beta2-AR, the BRET2 assay may be superior for secondary screening of agonists where a separation of full and partial agonists is needed and the ALPHAscreen cAMP assay may be preferred for primary screening of agonists where all receptor activating compounds are desired. PMID- 17114330 TI - A socioecological approach to improving mammography rates in a tribal community. AB - This article highlights the processes and intermediate outcomes of a pilot project to increase mammography rates of women in an American Indian tribe in New Mexico. Using a socioecological framework and principles of community-based participatory research, a community coalition was able to (a) bolster local infrastructure to increase access to mammography services; (b) build public health knowledge and skills among tribal health providers; (c) identify community specific knowledge, attitudes, and beliefs related to breast cancer; (d) establish interdependent partnerships among community health programs and between the tribe and outside organizations; and (e) adopt local policy initiatives to bolster tribal cancer control. These findings demonstrate the value of targeting a combination of individual, community, and environmental factors, which affect community breast cancer screening rates and incorporating cultural strengths and resources into all facets of a tribal health promotion intervention. PMID- 17114332 TI - Personal involvement of young people in HIV prevention campaign messages: the role of message format, culture, and gender. AB - To examine young people's reactions to and understanding of HIV prevention messages developed for MTV's global HIV prevention campaign Staying Alive, videotaped campaign materials were shown to focus group discussion (FGD) participants living in urban areas of Brazil, Kenya, Nepal, and Senegal. Responses related to "personal involvement" with the message were identified in the data from these FGDs and were examined in relationship to the emerging message themes, the message format (public service announcements [PSAs] vs. documentary), cultural context (site), and participant gender. Across groups, greater personal involvement (measured by personal connections, emotional reactions, and lessons learned) was found in responses about the documentary format compared to the PSA format. Exceptions were found for specific PSAs that were considered more relevant within specific gender or cultural contexts. Implications of findings for global campaigns were considered. PMID- 17114331 TI - Randomized trials on consider this, a tailored, internet-delivered smoking prevention program for adolescents. AB - The Internet may be an effective medium for delivering smoking prevention to children. Consider This, an Internet-based program, was hypothesized to reduce expectations concerning smoking and smoking prevalence. Group-randomized pretest posttest controlled trials were conducted in Australia (n = 2,077) and the United States (n = 1,234) in schools containing Grades 6 through 9. Australian children using Consider This reported reduced 30-day smoking prevalence. This reduction was mediated by decreased subjective norms. The amount of program exposure was low in many classes, but program use displayed a dose-response relationship with reduced smoking prevalence. American children only reported lower expectations for smoking in the future. Intervening to prevent smoking is a challenge, and this data suggest small benefits from an Internet-based program that are unlikely to be of practical significance unless increased by improved implementation. Implementation remains the major challenge to delivering interventions via the Internet, both for health educators and researchers. PMID- 17114333 TI - Attitudinal and relational factors predicting the use of solar water disinfection: a field study in Nicaragua. AB - Solar water disinfection (SODIS) is an uncomplicated and cheap technology providing individuals with safe drinking water by exposing water-filled plastic bottles to sunlight for 6 hours to kill waterborne pathogens. Two communities were visited, and 81 families (40 SODIS users and 41 nonusers) were interviewed. The relationship between several factors and the intention to use SODIS in the future and actual use were tested. The results showed that intention to use and actual use are mainly related to an overall positive attitude, intention to use is related to the use of SODIS by neighbors, and actual use is related to knowledge about SODIS; SODIS users reported a significantly lower incidence in diarrhea than SODIS nonusers. These results suggest that promotion activities should aim at creating a positive attitude, for example, by choosing a promoter that is able to inspire confidence in the new technology. PMID- 17114339 TI - Down syndrome candidate region 1 isoform 1 mediates angiogenesis through the calcineurin-NFAT pathway. AB - Down syndrome candidate region 1 (DSCR1) is one of more than 50 genes located in a region of chromosome 21 that has been implicated in Down syndrome. DSCR1 can be expressed as four isoforms, one of which, isoform 4 (DSCR1-4), has recently been found to be strongly induced by vascular endothelial growth factor A (VEGF A(165)) and to provide a negative feedback loop that inhibits VEGF-A(165)-induced endothelial cell proliferation in vitro and angiogenesis in vivo. We report here that another DSCR1 isoform, DSCR1-1L, is also up-regulated by VEGF-A(165) in cultured endothelial cells and is strongly expressed in several types of pathologic angiogenesis in vivo. In contrast to DSCR1-4, the overexpression of DSCR1-1L induced the proliferation and activation of the transcription factor NFAT in cultured endothelial cells and promoted angiogenesis in Matrigel assays in vivo, even in the absence of VEGF-A. Similarly, small interfering RNAs specific for DSCR1-1L and DSCR1-4 had opposing inhibitory and stimulatory effects, respectively, on these same functions. DSCR1-4 is thought to inhibit angiogenesis by inactivating calcineurin, thereby preventing activation and nuclear translocation of NFAT, a key transcription factor. In contrast, DSCR1-1L, regulated by a different promoter than DSCR1-4, activates NFAT and its proangiogenic activity is inhibited by cyclosporin, an inhibitor of calcineurin. In sum, DSCR1-1L, unlike DSCR1-4, potently activates angiogenesis and could be an attractive target for antiangiogenesis therapy. PMID- 17114340 TI - Lipocalin 2 antagonizes the proangiogenic action of ras in transformed cells. AB - Lipocalin 2 is an iron-binding secreted protein that converts embryonic kidney mesenchyme to epithelia. Previously, we reported that lipocalin 2 could revert 4T1-ras-transformed mesenchymal tumor cells to a more epithelial phenotype, increase E-cadherin expression, and suppress cell invasiveness in vitro and in vivo, indicating that lipocalin 2 is a metastasis suppressor. Here, we show that lipocalin 2 can suppress the ras-induced expression of vascular endothelial growth factor in 4T1 cells via down-regulation of ras mitogen-activated protein kinase and ras phosphatidylinositol-3-kinase signaling. In addition, the expression of thrombospondin-1 (an antiangiogenic molecule) was increased in tumors formed by 4T1-ras cells into which lipocalin 2 was stably introduced. Tumor angiogenesis, assessed via an intradermal tumor angiogenesis assay, was also suppressed by lipocalin 2. We also show that caveolin-1 is a critical mediator of this activity. These data provide new insights into the action of lipocalin 2 and raise the possibility that the administration of lipocalin 2 may be useful for inhibiting tumor angiogenesis, in addition to suppressing tumor metastasis, in cancers which show ras activation. PMID- 17114341 TI - Inhibition of matrilysin expression by antisense or RNA interference decreases lysophosphatidic acid-induced epithelial ovarian cancer invasion. AB - Our previous reports show that matrilysin [matrix metalloproteinase (MMP)-7] is overexpressed in epithelial ovarian cancer (EOC) and recombinant MMP-7 promotes EOC invasion in vitro. In the present study, we further evaluated the correlation of MMP-7 expression to EOC invasiveness and examined its role in lysophosphatidic acid (LPA)-induced invasion. By sense and antisense gene transfection in vitro, we show that overexpression of MMP-7 in all MMP-7 stably transfected DOV13 clones significantly enhanced their invasiveness, although MMP-7 antisense transfection caused a 91% decrease of MMP-7 expression (P < 0.01) and 87% decrease of invasion (P < 0.05) in geneticin (G418)-selected DOV13 clone P47-M7As-3 compared with vector-transfected control. As assessed by MMP-7 ELISA, LPA treatment at 10 to 80 micromol/L significantly stimulated the secretion of total MMP-7 in DOV13 conditioned medium (P < 0.01). In addition, LPA apparently induced the activation of MMP-7 in DOV13 cells as detected by gelatin zymography. In the antisense MMP-7 transfected DOV13 clone (P47-M7As-3), LPA-increased invasion was significantly decreased compared with vector control. Moreover, knocking down of MMP-7 by small interfering RNA also suppressed LPA-induced invasion in two EOC cell lines (DOV13 and R182). Altogether, our results show that MMP-7 expression is correlated with EOC invasiveness and LPA-induced MMP-7 secretion/activation may represent a new mechanism that facilitates ovarian cancer invasion besides the well-known induction of MT1-MMP-mediated proMMP-2 activation by LPA. PMID- 17114342 TI - Enhanced expression of LKB1 in breast cancer cells attenuates angiogenesis, invasion, and metastatic potential. AB - LKB1 (also known as STK11) is a recently identified tumor suppressor gene whose mutation can lead to Peutz-Jeghers syndrome, which is characterized by gastrointestinal polyps and cancers of different organ systems. Approximately 30% of sporadic breast cancer samples express low levels of LKB1. This suggests that the LKB1 gene may be related to the tumorigenesis of breast cancer. We reintroduced LKB1 into MDA-MB-435 breast cancer cells that lack the LKB1 gene to investigate how overexpression of LKB1 affects tumor invasiveness and metastasis. Overexpression of the LKB1 protein in breast cancer cells resulted in significant inhibition of in vitro invasion. In vivo, LKB1 expression reduced tumor growth in the mammary fat pad, microvessel density, and lung metastasis. LKB1 overexpression was associated with down-regulation of matrix metalloproteinase-2, matrix metalloproteinase-9, vascular endothelial growth factor, and basic fibroblast growth factor mRNA and protein levels. Overexpression of the LKB1 protein in human breast cancer is significantly associated with a decrease in microvessel density. Our results indicate that LKB1 plays a negative regulatory role in human breast cancer, a finding that may lead to a new therapeutic strategy. PMID- 17114343 TI - NR0B1 is required for the oncogenic phenotype mediated by EWS/FLI in Ewing's sarcoma. AB - A number of solid tumors, such as alveolar rhabdomyosarcoma, synovial sarcoma, and myxoid liposarcoma, are associated with recurrent translocation events that encode fusion proteins. Ewing's sarcoma is a pediatric tumor that serves as a prototype for this tumor class. Ewing's sarcomas usually harbor the (11;22)(q24;q12) translocation. The t(11;22) encodes the EWS/FLI fusion oncoprotein. EWS/FLI functions as an aberrant transcription factor, but the key target genes that are involved in oncogenesis are largely unknown. Although some target genes have been defined, many of these have been identified in heterologous model systems with uncertain relevance to the human disease. To understand the function of EWS/FLI and its targets in a more clinically relevant system, we used retroviral-mediated RNAi to "knock-down" the fusion protein in patient-derived Ewing's sarcoma cell lines. By combining transcriptional profiling data from three of these lines, we identified a conserved transcriptional response to EWS/FLI. The gene that was most reproducibly up regulated by EWS/FLI was NR0B1. NR0B1 is a developmentally important orphan nuclear receptor with no previously defined role in oncogenesis. We validated NR0B1 as an EWS/FLI-dysregulated gene and confirmed its expression in primary human tumor samples. Functional studies revealed that ongoing NR0B1 expression is required for the transformed phenotype of Ewing's sarcoma. These studies define a new role for NR0B1 in oncogenic transformation and emphasize the utility of analyzing the function of EWS/FLI in Ewing's sarcoma cells. PMID- 17114344 TI - The tumor suppressor KLF11 mediates a novel mechanism in transforming growth factor beta-induced growth inhibition that is inactivated in pancreatic cancer. AB - c-myc promoter silencing is a key step in epithelial cell growth inhibition by transforming growth factor beta (TGFbeta). During carcinogenesis, however, epithelial cells escape from c-myc repression and consequently become refractory to TGFbeta-mediated antiproliferation. Here, we assessed the role of the repressor, KLF11, in TGFbeta-induced growth inhibition in normal epithelial as well as pancreatic carcinoma cells. Endogenous KLF11 was stably down-regulated by RNA interference technology, and the functional consequences were studied by proliferation assays, reporter assays, DNA binding studies, and expression analyses. Coimmunoprecipitation and glutathione S-transferase pulldown assays were conducted to define KLF11-Smad3 interaction and U0126 was administered to examine the effects of the extracellular signal-regulated kinase (ERK)-mitogen activated protein kinase on complex formation and c-myc promoter binding of KLF11 and Smad3 in pancreatic cancer cells. In TGFbeta-stimulated normal epithelial cells, nuclear KLF11, in concert with Smad3, binds to and represses transcription from the core region of the TGFbeta-inhibitory element (TIE) of the c-myc promoter. Disruption of KLF11-Smad3 interaction or small interfering RNA-mediated knockdown of endogenous KLF11 strongly diminishes Smad3-TIE promoter binding and repression, and consequently impairs TGFbeta-mediated growth inhibition. In pancreatic cancer cells with oncogenic Ras mutations, hyperactive ERK counteracts TGFbeta-induced c-myc repression and growth inhibition through at least two mechanisms, i.e., via disruption of KLF11-Smad3 complex formation and through inhibition of KLF11-Smad3 binding to the TIE element. Together, these results suggest a central role for KLF11 in TGFbeta-induced c-myc repression and antiproliferation and identifies a novel mechanism through which ERK signaling antagonizes the tumor suppressor activities of TGFbeta in pancreatic cancer cells with oncogenic Ras mutations. PMID- 17114345 TI - MUC1 oncoprotein functions in activation of fibroblast growth factor receptor signaling. AB - Activation of the fibroblast growth factor (FGF) receptor 3 (FGFR3) has been linked to the development of human cancers by mechanisms that are not well understood. The MUC1 oncoprotein is aberrantly overexpressed by certain hematologic malignancies and most human carcinomas. The present studies show that MUC1 associates with FGFR3. Stimulation of cells with FGF1 increased the interaction between MUC1 and FGFR3. FGF1 stimulation also induced c-Src-dependent tyrosine phosphorylation of the MUC1 cytoplasmic domain on a YEKV motif. FGF1 induced tyrosine phosphorylation of MUC1 was associated with increased binding of MUC1 to beta-catenin and targeting of MUC1 and beta-catenin to the nucleus. FGF1 also induced binding of MUC1 to the heat shock protein 90 (HSP90) chaperone by a mechanism dependent on phosphorylation of the YEKV motif. Notably, beta-catenin and HSP90 compete for binding to the MUC1 cytoplasmic domain, indicating that MUC1 forms mutually exclusive complexes with these proteins. The results also show that inhibition of HSP90 with geldanamycin or 17-(allylamino)-17 demethoxygeldanamycin attenuates FGF1-induced binding of MUC1 to HSP90 and targeting of MUC1 to the mitochondrial outer membrane. These findings indicate that FGF1 induces phosphorylation of MUC1 on YEKV and thereby activates two distinct pathways: (a) nuclear localization of MUC1 and beta-catenin and (b) delivery of MUC1 to mitochondria by HSP90. PMID- 17114346 TI - Tuberin nuclear localization can be regulated by phosphorylation of its carboxyl terminus. AB - Tuberin, the tuberous sclerosis 2 (TSC2) gene product, has been identified as a tumor suppressor protein genetically implicated in the pathology of tuberous sclerosis and the female-specific lung disease lymphangioleiomyomatosis. Tuberin and its predominant cytoplasmic binding partner hamartin have been shown to complex with a variety of intracellular signaling regulators and affect the processes of protein translation, cellular proliferation, cellular migration, and cellular transcription. In previous studies, we have presented evidence for tuberin binding to the calcium-dependent intracellular signaling protein calmodulin (CaM), overlap of tuberin CaM binding domain with a binding domain for estrogen receptor alpha, and the phosphorylation-associated nuclear localization of tuberin. In the study presented here, we expand our findings on the mechanism of tuberin nuclear localization to show that the CaM-estrogen receptor-alpha binding domain of tuberin can also serve as a tuberin nuclear localization sequence. Furthermore, we identify an Akt/p90 ribosomal S6 kinase-1 phosphorylation site within the carboxyl terminus of tuberin that can regulate tuberin nuclear localization and significantly affect the ability of tuberin to modulate estrogen genomic signaling events. These findings suggest a link between tuberin nuclear localization and a variety of intracellular signaling events that have direct implications with respect to the role of tuberin in the pathology of tuberous sclerosis and lymphangioleiomyomatosis. PMID- 17114347 TI - First evidence of higher female recombination in a species with temperature dependent sex determination: the saltwater crocodile. AB - The first evidence of genetic linkage and sex-specific recombination in the order Crocodylia is reported. This study was conducted using a resource pedigree of saltwater crocodiles consisting of 16 known-breeding pairs (32 adults) and 101 juveniles. A total of 21 microsatellite loci were available for analysis. Ten of the 21 loci showed linkage with 4 linkage groups: 3 pairwise (Cj131/Cj127, CUD68/Cj101, and Cj107/Cp10) and 1 four-locus (Cj122, CUD78, Cj16, and Cj104) being found. Linkage analysis on the 21 loci revealed evidence of sex-specific differences in recombination rates. All 5 nonzero interlocus intervals were longer in females than in males, with the 4-loci linkage group 3-fold longer in females than in males (41.63 cM and 14.1 cM, respectively). This is the first report of sex-specific recombination rates in a species that exhibits temperature dependent sex determination. PMID- 17114348 TI - SND1, a NAC domain transcription factor, is a key regulator of secondary wall synthesis in fibers of Arabidopsis. AB - Secondary walls in fibers and tracheary elements constitute the most abundant biomass produced by plants. Although a number of genes involved in the biosynthesis of secondary wall components have been characterized, little is known about the molecular mechanisms underlying the coordinated expression of these genes. Here, we demonstrate that the Arabidopsis thaliana NAC (for NAM, ATAF1/2, and CUC2) domain transcription factor, SND1 (for secondary wall associated NAC domain protein), is a key transcriptional switch regulating secondary wall synthesis in fibers. We show that SND1 is expressed specifically in interfascicular fibers and xylary fibers in stems and that dominant repression of SND1 causes a drastic reduction in the secondary wall thickening of fibers. Ectopic overexpression of SND1 results in activation of the expression of secondary wall biosynthetic genes, leading to massive deposition of secondary walls in cells that are normally nonsclerenchymatous. In addition, we have found that SND1 upregulates the expression of several transcription factors that are highly expressed in fibers during secondary wall synthesis. Together, our results reveal that SND1 is a key transcriptional activator involved in secondary wall biosynthesis in fibers. PMID- 17114349 TI - Arabidopsis chromatin-associated HMGA and HMGB use different nuclear targeting signals and display highly dynamic localization within the nucleus. AB - In plants, the chromatin-associated high mobility group (HMG) proteins occur in two subfamilies termed HMGA and HMGB. The HMGA proteins are characterized by the presence of four AT-hook DNA binding motifs, and the HMGB proteins contain an HMG box DNA binding domain. As architectural factors, the HMG proteins appear to be involved in the regulation of transcription and other DNA-dependent processes. We have examined the subcellular localization of Arabidopsis thaliana HMGA, HMGB1, and HMGB5, revealing that they localize to the cell nucleus. They display a speckled distribution pattern throughout the chromatin of interphase nuclei, whereas none of the proteins associate with condensed mitotic chromosomes. HMGA is targeted to the nucleus by a monopartite nuclear localization signal, while efficient nuclear accumulation of HMGB1/5 requires large portions of the basic N terminal part of the proteins. The acidic C-terminal domain interferes with nucleolar targeting of HMGB1. Fluorescence recovery after photobleaching experiments revealed that HMGA and HMGB proteins are extremely dynamic in the nucleus, indicating that they bind chromatin only transiently before moving on to the next site, thereby continuously scanning the genome for targets. By contrast, the majority of histone H2B is basically immobile within the nucleus, while linker histone H1.2 is relatively mobile. PMID- 17114350 TI - Arabidopsis SLIM1 is a central transcriptional regulator of plant sulfur response and metabolism. AB - Sulfur is an essential macronutrient required for plant growth. To identify key transcription factors regulating the sulfur assimilatory pathway, we screened Arabidopsis thaliana mutants using a fluorescent reporter gene construct consisting of the sulfur limitation-responsive promoter of the SULTR1;2 sulfate transporter and green fluorescent protein as a background indicator for monitoring plant sulfur responses. The isolated mutant, sulfur limitation1 (slim1), was unable to induce SULTR1;2 transcripts under low-sulfur (-S) conditions. Mutations causing the sulfur limitation responseless phenotypes of slim1 were identified in an EIL family transcription factor, ETHYLENE INSENSITIVE3-LIKE3 (EIL3), whose functional identity with SLIM1 was confirmed by genetic complementation. Sulfate uptake and plant growth on -S were significantly reduced by slim1 mutations but recovered by overexpression of SLIM1. SLIM1 functioned as a central transcriptional regulator, which controlled both the activation of sulfate acquisition and degradation of glucosinolates under -S conditions. Metabolite analysis indicated stable accumulation of glucosinolates in slim1 mutants, even under -S conditions, particularly in the molecular species with methylsulfinylalkyl side chains beneficial to human health. Overexpression of SLIM1 and its rice (Oryza sativa) homologs, but no other EIL genes of Arabidopsis, restored the sulfur limitation responseless phenotypes of slim1 mutants, suggesting uniqueness of the SLIM1/EIL3 subgroup members as sulfur response regulators. PMID- 17114351 TI - Protein polyubiquitination plays a role in basal host resistance of barley. AB - To study protein ubiquitination pathways in the interaction of barley (Hordeum vulgare) with the powdery mildew fungus (Blumeria graminis), we measured protein turnover and performed transient-induced gene silencing (TIGS) of ubiquitin and 26S proteasome subunit encoding genes in epidermal cells. Attack by B. graminis hyperdestabilized a novel unstable green fluorescent protein fusion that contains a destabilization domain of a putative barley 1-aminocyclopropane-1-carboxylate synthase, suggesting enhanced protein turnover. Partial depletion of cellular ubiquitin levels by TIGS induced extreme susceptibility of transformed cells toward the appropriate host pathogen B. graminis f. sp hordei, whereas papilla based resistance to the nonhost pathogen B. graminis f. sp tritici and host resistance mediated by the mlo gene (for mildew resistance locus O) remained unaffected. Cells were rescued from TIGS-induced ubiquitin depletion by synthetic genes encoding wild-type or mutant barley monoubiquitin proteins. The strongest rescue was from a gene encoding a K63R mutant form of ubiquitin blocked in several ubiquitination pathways while still allowing Lys-48-dependent polyubiquitination required for proteasomal protein degradation. Systematic RNA interference of 40 genes encoding all 17 subunits of the proteasome 19S regulatory particle failed to induce hypersusceptibility against B. graminis f. sp hordei. This suggests a role for Lys-48-linked protein polyubiquitination, which is independent from the proteasome pathway, in basal host defense of barley. PMID- 17114352 TI - Lack of the light-harvesting complex CP24 affects the structure and function of the grana membranes of higher plant chloroplasts. AB - The photosystem II (PSII) light-harvesting antenna in higher plants contains a number of highly conserved gene products whose function is unknown. Arabidopsis thaliana plants depleted of one of these, the CP24 light-harvesting complex, have been analyzed. CP24-deficient plants showed a decrease in light-limited photosynthetic rate and growth, but the pigment and protein content of the thylakoid membranes were otherwise almost unchanged. However, there was a major change in the macroorganization of PSII within these membranes; electron microscopy and image analysis revealed the complete absence of the C(2)S(2)M(2) light-harvesting complex II (LHCII)/PSII supercomplex predominant in wild-type plants. Instead, only C(2)S(2) supercomplexes, which are deficient in the LHCIIb M-trimers, were found. Spectroscopic analysis confirmed the disruption of the wild-type macroorganization of PSII. It was found that the functions of the PSII antenna were disturbed: connectivity between PSII centers was reduced, and maximum photochemical yield was lowered; rapidly reversible nonphotochemical quenching was inhibited; and the state transitions were altered kinetically. CP24 is therefore an important factor in determining the structure and function of the PSII light-harvesting antenna, providing the linker for association of the M trimer into the PSII complex, allowing a specific macroorganization that is necessary both for maximum quantum efficiency and for photoprotective dissipation of excess excitation energy. PMID- 17114353 TI - Insights into nuclear organization in plants as revealed by the dynamic distribution of Arabidopsis SR splicing factors. AB - Serine/arginine-rich (SR) proteins are splicing regulators that share a modular structure consisting of one or two N-terminal RNA recognition motif domains and a C-terminal RS-rich domain. We investigated the dynamic localization of the Arabidopsis thaliana SR protein RSZp22, which, as we showed previously, distributes in predominant speckle-like structures and in the nucleolus. To determine the role of RSZp22 diverse domains in its nucleolar distribution, we investigated the subnuclear localization of domain-deleted mutant proteins. Our results suggest that the nucleolar localization of RSZp22 does not depend on a single targeting signal but likely involves different domains/motifs. Photobleaching experiments demonstrated the unrestricted dynamics of RSZp22 between nuclear compartments. Selective inhibitor experiments of ongoing cellular phosphorylation influenced the rates of exchange of RSZp22 between the different nuclear territories, indicating that SR protein mobility is dependent on the phosphorylation state of the cell. Furthermore, based on a leptomycin B- and fluorescence loss in photobleaching-based sensitive assay, we suggest that RSZp22 is a nucleocytoplasmic shuttling protein. Finally, with electron microscopy, we confirmed that RSp31, a plant-specific SR protein, is dynamically distributed in nucleolar cap-like structures upon phosphorylation inhibition. Our findings emphasize the high mobility of Arabidopsis SR splicing factors and provide insights into the dynamic relationships between the different nuclear compartments. PMID- 17114354 TI - The transcription factors WRKY11 and WRKY17 act as negative regulators of basal resistance in Arabidopsis thaliana. AB - Transcription factors are believed to play a pivotal role in the activation and fine-tuning of plant defense responses, but little is known about the exact function of individual transcription factors in this process. We analyzed the role of the IId subfamily of WRKY transcription factors in the regulation of basal resistance to Pseudomonas syringae pv tomato (Pst). The expression of four members of the subfamily was induced upon challenge with virulent and avirulent strains of Pst. Mutant analyses revealed that loss of WRKY11 function increased resistance toward avirulent and virulent Pst strains and that resistance was further enhanced in wrky11 wrky17 double mutant plants. Thus, WRKY11 and WRKY17 act as negative regulators of basal resistance to Pst. Genome-wide expression analysis and expression studies of selected genes in single and double mutants demonstrated that both transcription factors modulate transcriptional changes in response to pathogen challenge. Depending on the target gene, WRKY11 and WRKY17 act either specifically or in a partially redundant manner. We demonstrate complex cross-regulation within the IId WRKY subfamily and provide evidence that both WRKY transcription factors are involved in the regulation of Pst-induced jasmonic acid-dependent responses. These results provide genetic evidence for the importance of WRKY11 and WRKY17 in plant defense. PMID- 17114355 TI - Subcellular trafficking of the Arabidopsis auxin influx carrier AUX1 uses a novel pathway distinct from PIN1. AB - The directional flow of the plant hormone auxin mediates multiple developmental processes, including patterning and tropisms. Apical and basal plasma membrane localization of AUXIN-RESISTANT1 (AUX1) and PIN-FORMED1 (PIN1) auxin transport components underpins the directionality of intercellular auxin flow in Arabidopsis thaliana roots. Here, we examined the mechanism of polar trafficking of AUX1. Real-time live cell analysis along with subcellular markers revealed that AUX1 resides at the apical plasma membrane of protophloem cells and at highly dynamic subpopulations of Golgi apparatus and endosomes in all cell types. Plasma membrane and intracellular pools of AUX1 are interconnected by actin dependent constitutive trafficking, which is not sensitive to the vesicle trafficking inhibitor brefeldin A. AUX1 subcellular dynamics are not influenced by the auxin influx inhibitor NOA but are blocked by the auxin efflux inhibitors TIBA and PBA. Furthermore, auxin transport inhibitors and interference with the sterol composition of membranes disrupt polar AUX1 distribution at the plasma membrane. Compared with PIN1 trafficking, AUX1 dynamics display different sensitivities to trafficking inhibitors and are independent of the endosomal trafficking regulator ARF GEF GNOM. Hence, AUX1 uses a novel trafficking pathway in plants that is distinct from PIN trafficking, providing an additional mechanism for the fine regulation of auxin transport. PMID- 17114356 TI - Psb27, a cyanobacterial lipoprotein, is involved in the repair cycle of photosystem II. AB - Photosystem II (PSII) performs one of the key reactions on our planet: the light driven oxidation of water. This fundamental but very complex process requires PSII to act in a highly coordinated fashion. Despite detailed structural information on the fully assembled PSII complex, the dynamic aspects of formation, processing, turnover, and degradation of PSII with at least 19 subunits and various cofactors are still not fully understood. Transient complexes are especially difficult to characterize due to low abundance, potential heterogeneity, and instability. Here, we show that Psb27 is involved in the assembly of the water-splitting site of PSII and in the turnover of the complex. Psb27 is a bacterial lipoprotein with a specific lipid modification as shown by matrix-assisted laser-desorption ionization time of flight mass spectrometry. The combination of HPLC purification of four different PSII subcomplexes and (15)N pulse label experiments revealed that lipoprotein Psb27 is part of a preassembled PSII subcomplex that represents a distinct intermediate in the repair cycle of PSII. PMID- 17114357 TI - A constitutive shade-avoidance mutant implicates TIR-NBS-LRR proteins in Arabidopsis photomorphogenic development. AB - In plants, light signals caused by the presence of neighbors accelerate stem growth and flowering and induce a more erect position of the leaves, a developmental strategy known as shade-avoidance syndrome. In addition, mutations in the photoreceptors that mediate shade-avoidance responses enhance disease susceptibility in Arabidopsis thaliana. Here, we describe the Arabidopsis constitutive shade-avoidance1 (csa1) mutant, which shows a shade-avoidance phenotype in the absence of shade and enhanced growth of a bacterial pathogen. The csa1 mutant has a T-DNA inserted within the second exon of a Toll/Interleukin1 receptor-nucleotide binding site-leucine-rich repeat (TIR-NBS LRR) gene, which leads to the production of a truncated mRNA. Arabidopsis plants transformed with the truncated TIR-NBS-LRR gene recapitulate the mutant phenotype, indicating that csa1 is a dominant-negative mutation that interferes with phytochrome signaling. TIR-NBS-LRR proteins have been implicated in defense responses in plants. RPS4, the closest homolog of CSA1, confers resistance to Pseudomonas syringae and complements the csa1 mutant phenotype, indicating that responses to pathogens and neighbors share core-signaling components in Arabidopsis. In Drosophila melanogaster and Caenorhabditis elegans, TIR domain proteins are implicated in both development and immunity. Thus, the dual role of the TIR domain is conserved across kingdoms. PMID- 17114359 TI - Celebrating 60 years of Thorax. PMID- 17114358 TI - A comparison of transcriptomic and metabonomic technologies for identifying biomarkers predictive of two-year rodent cancer bioassays. AB - Two-year rodent bioassays play a central role in evaluating the carcinogenic potential of both commercial products and environmental contaminants. The bioassays are expensive and time consuming, requiring years to complete and costing $2-4 million. In this study, we compare transcriptomic and metabonomic technologies for discovering biomarkers that can efficiently and economically identify chemical carcinogens without performing a standard two-year rodent bioassay. Animals were exposed subchronically to two chemicals (one genotoxic and one nongenotoxic) that were positive for lung and liver tumors in a standard two year bioassay, two chemicals that were negative, and two control groups. Microarray analysis performed on liver and lung tissues identified multiple biomarkers in each tissue that could discriminate between carcinogenic and noncarcinogenic treatments. The discriminating biomarkers shared a common expression profile among carcinogenic treatments despite different genotoxicity categories and potential modes of action, suggesting that they reflect underlying cellular changes in the transition toward neoplasia. Statistical classification analysis exhibited 100% accuracy in both tissues when the number of genes was less than 5000. Additional genes reduced the predictive accuracy of the model. Serum samples were analyzed by 1H nuclear magnetic resonance (NMR) spectroscopy, and chemical-specific metabolites were removed from the spectra. The statistical classification analysis of the endogenous serum metabolites showed relatively low predictive accuracy with few metabolites in the model, but the accuracy increased to a maximum of 94% when all metabolites were added. These results suggest that individual endogenous metabolites are relatively poor biomarkers, but the metabolite profile as a whole is altered following carcinogen treatment. PMID- 17114368 TI - Carcinoma of the bronchus 60 years later. PMID- 17114369 TI - Emphysema in COPD: consequences and causes. PMID- 17114370 TI - Underdiagnosed chronic obstructive pulmonary disease in England: new country, same story. PMID- 17114371 TI - Alpha-1-antitrypsin augmentation treatment: does one size fit all? PMID- 17114373 TI - Lymphocytic interstitial pneumonia associated with common variable immunodeficiency resolved with intravenous immunoglobulins. AB - Lymphocytic interstitial pneumonia (LIP) is a rare form of interstitial lung disease. A few case reports have described an association with common variable immunodeficiency (CVID). Corticosteroids are usually used to treat symptomatic patients but their efficacy has never been studied in a controlled trial. We describe a patient with LIP and CVID who was treated monthly with intravenous immunoglobulins (IVIG) without steroids. The patient improved dramatically. We believe that, in selected cases of LIP and immunodeficiency, IVIG given monthly should be considered as the only treatment without adding steroids. PMID- 17114374 TI - Still awaiting a useful tool for predicting severe CAP. PMID- 17114375 TI - Bacterial co-infection and interpretation of immunological data from BAL fluid specimens in severe RSV bronchiolitis. PMID- 17114376 TI - Concurrence of sarcoidosis and lung cancer: a diagnostic dilemma. PMID- 17114377 TI - British Thoracic Society Winter Meeting 2006, London, England, 6-8 December 2006. Abstracts. PMID- 17114380 TI - Herd health planning: farmers' perceptions in relation to lameness and mastitis. AB - Between December 2002 and December 2003, the herd health planning activities on 61 dairy farms in the uk were compared with several measures of lameness and mastitis. Lameness had been reported as a problem in 53 of the herds directly by the farm and in the other eight by the nominating local veterinary practice; 54 of the farms also reported having a mastitis problem. Fifty-three (87 per cent) of the farms had some form of written herd health plan, of which 21 (40 per cent) had been in place for 12 months or less. All the farms were recording mastitis in some way, although 38 (62 per cent) of the farmers did not review these records and only four retained the results of a comprehensive record review. Farms defined as having a high incidence of mastitis were more likely to be reviewing their health records, but farms defined as having a high prevalence of lameness in a sentinel group of early lactation heifers were less likely to be reviewing their health records. PMID- 17114381 TI - Effects of two doses of buprenorphine four or six hours apart on nociceptive thresholds, pain and sedation in dogs after castration. AB - Twenty-eight dogs were randomly allocated into two groups. They were premedicated with either 10 or 20 microg/kg buprenorphine and 0.05 mg/kg acepromazine administered intramuscularly, and then anaesthetised with intravenous thiopentone to effect and maintained with isoflurane in 100 per cent oxygen. The dogs underwent routine castration, and a second dose of 10 microg/kg buprenorphine was administered four hours after the first or 20 microg/kg six hours after the first dose. Levels of pain and sedation were scored on a visual analogue scale and in terms of the dogs' requirement for rescue analgesia, and mechanical nociceptive thresholds were measured at the hock and wound at premedication and one, two, three, four, five, six, seven, 10 and 21 to 22 hours later. Pain scores were low in both groups, with a trend for lower scores in the high dose group; administration of the second dose of buprenorphine further decreased the pain scores. Buprenorphine produced good preoperative sedation and the level of sedation decreased over time after surgery. Administration of the second high dose of buprenorphine did not increase the level of sedation. Both doses of buprenorphine prevented hyperalgesia at the wound and hock postoperatively. Three dogs given the low dose and one dog given the high dose required rescue analgesia with carprofen. PMID- 17114382 TI - In vitro study of the effectiveness of different dressings for debriding fibrin in blood clots from horses. AB - Franz diffusion cells containing uniformly dehydrated equine blood clots to simulate fibrinous eschar were used to measure the rate of debridement of fibrin by novel and traditional wound dressings, under standardised conditions of temperature, pH and humidity. Significant increases in protein breakdown occurred within 24 hours with all the dressings, but not thereafter. In general, dressings hydrated in normal saline were better as debriding agents than dressings hydrated in water. Autolytic debriding agents were 47 per cent more effective than chemical debriding agents; specifically, hydrofibre and gauze dressings hydrated in saline broke down more than 3500 microg/ml of protein whereas dressings impregnated with proteolytic enzymatic agents digested less than 1400 microg/ml. PMID- 17114383 TI - Comparison of three ELISAs for the diagnosis of porcine reproductive and respiratory syndrome. PMID- 17114384 TI - Coexistence of pituitary adenocarcinoma and intraocular melanoma in a sheep. PMID- 17114385 TI - Mycotic blepharitis associated with Fusarium species in a Eurasian eagle owl (Bubo bubo hispanus). PMID- 17114386 TI - Efficacy of vaccination against canine parvovirus. PMID- 17114387 TI - Ovariohysterectomy versus ovariectomy. PMID- 17114388 TI - West Nile virus in horses. PMID- 17114391 TI - MRSA in companion animals. PMID- 17114392 TI - Vets' voice needed in modern farming. PMID- 17114393 TI - Interactions between the neural regulation of stress and aggression. AB - Socially aggressive interaction is stressful. What is more, social aggression is stressful for both dominant and subordinate animals. Much of the neurocircuitry for stress and aggression overlap. The pattern of neurochemical and hormonal events stimulated by social interaction make it clear that subtle differences in this pattern of response distinguish social rank. The neurotransmitter serotonin (5-HT) responds rapidly to stress, and also appears to play the most important role for inhibitory regulation of aggressive interactions. In addition, the adrenocortical/interrenal steroid hormones corticosterone and cortisol are responsive to stress and influence aggression. However, while 5-HT and glucocorticoids can both be inhibitory to aggression, the relationship between 5 HT and glucocorticoids is not straightforward, and much of the distinctions in function depend upon timing. Neither is inhibitory during the early stressful phase of aggression. This transmitter-hormone combination follows and influences a four-stage functional pattern of effect: (1) predisposed (positively or negatively) toward aggression, (2) motivated toward behavior, (3) responsive to stress (including aggression) and passively allowing aggression, and finally (4) chronically applied 5-HT and glucocorticoids inhibit aggression. PMID- 17114394 TI - Fooling a freshwater fish: how dietary salt transforms the rainbow trout gill into a seawater gill phenotype. AB - Numerous fish species, including rainbow trout (Oncorhynchus mykiss), are able to inhabit both freshwater and seawater and routinely migrate between the two environments. One of the most critical adjustments allowing such successful migrations is a remodelling of the gill in which a suite of morphological and molecular changes ensure optimal function in the face of reversing requirements for salt and water balance. The remodelling leads to specific freshwater and seawater gill phenotypes that are readily identified by the orientation and/or quantities of specific ion transporters and the presence or absence of specific cell types. The proximate cues promoting gill phenotypic plasticity are unknown. Here, by assessing the consequences of a salt-enriched diet (in the absence of any changes in external salinity) in the freshwater rainbow trout, we demonstrate that internal salt loading alone, is able to induce various elements of the seawater gill phenotype. Specifically, we show upregulation of three ion transport genes, cystic fibrosis transmembrane conductance regulator (CFTR), Na(+)/K(+)/2Cl(-) co-transporter (NKCC1) and Na(+)/K(+)-ATPase, which are essential for ionic regulation in seawater, and the appearance of chloride cell accessory cell complexes, which are normally restricted to fish inhabiting seawater. These data provide compelling evidence that gill remodelling during migration from freshwater to seawater may involve sensing of elevated levels of internal salt. PMID- 17114395 TI - A comparison of visual and haltere-mediated feedback in the control of body saccades in Drosophila melanogaster. AB - The flight trajectories of fruit flies consist of straight flight segments interspersed with rapid turns called body saccades. Although the saccades are stereotyped, it is not known whether their brief time course is due to a feed forward (predetermined) motor program or due to feedback from sensory systems that are reflexively activated by the rapid rotation. Two sensory modalities, the visual system and the mechanosensory halteres, are likely sources of such feedback because they are sensitive to angular velocities within the range experienced during saccades. Utilizing a magnetic tether in which flies are fixed in space but free to rotate about their yaw axis, we systematically manipulated the feedback from the visual and haltere systems to test their role in determining the time course of body saccades. We found that altering visual feedback had no significant effect on the dynamics of saccades, whereas increasing and decreasing the amount of haltere-mediated feedback decreased and increased saccade amplitude, respectively. In other experiments, we altered the aerodynamic surface of the wings such that the flies had to actively modify their wing-stroke kinematics to maintain straight flight on the magnetic tether. Flies exhibit such modification, but the control is compromised in the dark, indicating that the visual system does provide feedback for flight stability at lower angular velocities, to which the haltere system is less sensitive. Cutting the wing surface disrupted the time course of the saccades, indicating that although flies employ sensory feedback to modulate saccade dynamics, it is not precise or fast enough to compensate for large changes in wing efficacy. PMID- 17114396 TI - Jumping performance of froghopper insects. AB - The kinematics of jumping in froghopper insects were analysed from high speed sequences of images captured at rates up to 8000 s(-1). In a jump, the attitude of the body is set by the front and middle legs, and the propulsion is delivered by rapid and synchronous movements of the hind legs that are 1.5 times longer than the other legs, but are only about half the length of the body and represent just 2% of the body mass. The wings are not moved and the front and middle legs may be raised off the ground before take-off. The hind legs are first cocked by a slow levation of the trochantera about the coxae so that the femora are pressed against the ventral, indented wall of the thorax, with the femoro-tibial joints tucked between the middle legs and body. Only the tips of the hind tarsi are in contact with the ground. In this position, the hind legs stay motionless for 1-2 s. Both trochantera are then synchronously and rapidly depressed about the coxae at rotational velocities of 75 500 deg. s(-1) and the tibiae extended, to launch a jump that in Philaenus reaches a height of 700 mm, or 115 body lengths. In the best jumps by Philaenus, take-off occurs within 0.875 ms of the start of movements of the hind legs at a peak velocity of 4.7 m s(-1) and involves an acceleration of 5400 m s(-2), equivalent to 550 times gravity. This jumping performance requires an energy output of 136 microJ, a power output of 155 mW and exerts a force of 66 mN. PMID- 17114397 TI - Morphology and action of the hind leg joints controlling jumping in froghopper insects. AB - The morphology and movements of key joints of the hind legs that generate the rapid jumping of froghoppers were analysed. The movements of an individual hind leg during a jump occur in three phases. First, the trochanter is slowly levated about the coxa so that the femur moves anteriorly and engages with a lateral protrusion on the coxa. Second, both hind legs are held in this fully levated (cocked) position without moving for a few seconds. Third, both hind legs depress and extend completely in less than 1 ms. The critical, power-generating movement underlying a jump is the rapid and simultaneous depression of the trochantera about the coxae. The lever arm of the hind trochanteral depressor muscle is smallest at the cocked position, but does not appear to go over the centre of the pivot. It then increases to a maximum after some 80 degrees of depression movement. By contrast, the lever arm of the trochanteral levator tendon is similar over the range of joint movements and is exceeded by that of the depressor only after 40 degrees of depression. Three prominent arrays of hairs on the trochantin, coxa and trochanter are appropriately positioned to act as proprioceptors signalling key movements in jumping. In the fully levated position, a protrusion on the dorsal, proximal surface of a hind femur engages with a protrusion from the ventral and lateral part of a coxa. These structures are not present on the front and middle legs. Both protrusions are covered with a dense array of small projections (microtrichia) that both increase the surface area and may interlock with each other. To depress rapidly in a jump these protrusions must disengage. If the hind leg of a dead froghopper is forcibly levated, it will lock in its cocked position, from which it can depress rapidly by movement of the coxo-trochanteral joint and disengagement of the femoral and coxal protrusions. A prominent click sound occurs at the start of a jump that results either from the initial movements of the coxo-trochanteral joint, or from the disengagement of the microtrichia on the coxa and femur. Larval Philaenus, which do not jump, lack a femoral protrusion and have no microtrichia in equivalent positions on either the coxa or femur. PMID- 17114398 TI - P-type Na+/K+-ATPase and V-type H+-ATPase expression patterns in the osmoregulatory organs of larval and adult mosquito Aedes aegypti. AB - This study describes the expression patterns of P-type Na(+)/K(+)-ATPase and V type H(+)-ATPase in the larval and adult forms of the mosquito Aedes aegypti and provides insight into their relative importance in ion transport function of key osmoregulatory organs. RT-PCR assays indicate that, at the level of the gene, both ATPases are expressed in all of the osmoregulatory tissues of larvae (midgut, Malpighian tubules, rectum and anal papillae) and adults (stomach, Malpighian tubules, anterior hindgut and rectum). Immunohistochemical studies determined that both ATPases are present in high levels in all the relevant organs, with the exception of the larval rectum (P-type Na(+)/K(+)-ATPase only). In larval gastric caeca, ATPase location corresponds to the secretory (basal P type Na(+)/K(+)-ATPase, apical V-type H(+)-ATPase) and ion-transporting (V-type H(+)-ATPase on both membranes) regions as previously described. The two ATPases switch membrane location along the length of the larval midgut, indicating three possible regionalizations, whereas the adult stomach has uniform expression of basolateral P-type Na(+)/K(+)-ATPase and apical V-type H(+)-ATPase in each cell. In both larval and adult Malpighian tubules, the distal principal cells exhibit high expression levels of V-type H(+)-ATPase (apically and cytoplasmically) whereas P-type Na(+)/K(+)-ATPase is highly expressed in stellate cells found only in the distal two-thirds of each tubule. By contrast, the proximal principal cells express both P-type Na(+)/K(+)-ATPase (basal) and V-type H(+)-ATPase (apical). These results suggest a functional segregation along the length of the Malpighian tubules based on cell type and region. P-type Na(+)/K(+)-ATPase is the only pump apparent in the larval rectum whereas in the larval anal papillae and the adult hindgut (including the anterior hindgut and rectum with rectal pads), P type Na(+)/K(+)-ATPase and V-type H(+)-ATPase localize to the basal and apical membranes, respectively. We discuss our findings in light of previous physiological and morphological studies and re-examine our current models of ion transport in these two developmental stages of mosquitoes that cope with disparate osmoregulatory challenges. PMID- 17114399 TI - Sensorimotor control during isothermal tracking in Caenorhabditis elegans. AB - In order to purposefully navigate their environments, animals rely on precise coordination between their sensory and motor systems. The integrated performance of circuits for sensorimotor control may be analyzed by quantifying an animal's motile behavior in defined sensory environments. Here, we analyze the ability of the nematode C. elegans to crawl isothermally in spatial thermal gradients by quantifying the trajectories of individual worms responding to defined spatiotemporal thermal gradients. We show that sensorimotor control during isothermal tracking may be summarized as a strategy in which the worm changes the curvature of its propulsive undulations in response to temperature changes measured at its head. We show that a concise mathematical model for this strategy for sensorimotor control is consistent with the exquisite stability of the worm's isothermal alignment in spatial thermal gradients as well as its more complex trajectories in spatiotemporal thermal gradients. PMID- 17114400 TI - Lipid remodeling in wild and selectively bred hard clams at low temperatures in relation to genetic and physiological parameters. AB - A temperature decrease usually induces an ordering effect in membrane phospholipids, which can lead to membrane dysfunction. Poikilotherms inhabiting eurythermal environments typically counteract this temperature effect by remodeling membrane lipids as stipulated in the homeoviscous adaptation theory (HVA). Hard clams, Mercenaria mercenaria, can suffer high overwintering mortalities in the Gulf of St Lawrence, Canada. The selectively bred M. mercenaria var. notata can have higher overwintering mortalities than the wild species, thus suggesting that the two varieties have different degrees of adaptation to low temperatures. The objective of this study was to investigate the changes in lipid composition of soft tissues in wild and selected hard clams in relation to their metabolic and genetic characteristics. Clams were placed at the northern limit of their distribution from August 2003 to May 2004; they were exposed to a gradual temperature decrease and then maintained at <0 degrees C for 3.5 months. This study is the first to report a major remodeling of lipids in this species as predicted by HVA; this remodeling involved a sequential response of the phospholipid to sterol ratio as well as in levels of 22:6n-3 and non methylene interrupted dienoic fatty acids. Hard clams showed an increase in 20:5n 3 as temperature decreased, but this was not maintained during overwintering, which suggests that 20:5n-3 may have been used for eicosanoid biosynthesis as a stress response to environmental conditions. Selectively bred hard clams were characterized by a higher metabolic demand and a deviation from Hardy-Weinberg equilibrium at several genetic loci due to a deficit in heterozygote frequency compared with wild clams, which is believed to impose additional stress and render these animals more vulnerable to overwintering mortality. Finally, an intriguing finding is that the lower metabolic requirements of wild animals coincide with a lower unsaturation index of their lipids, as predicted by Hulbert's theory of membranes as pacemakers of metabolism. PMID- 17114401 TI - Neural control of the velum in larvae of the gastropod, Ilyanassa obsoleta. AB - Larval molluscs commonly use ciliated vela to swim and feed. In this study we used immunohistochemistry to demonstrate innervation of velar cilia and muscles by monoaminergic and peptidergic fibres in the caenogastropod, Ilyanassa obsoleta. Photoelectric recordings from pre-oral cilia on isolated pieces of velum revealed that serotonin increased, whereas catecholamines (dopamine and norepinephrine) decreased beat frequency at concentrations of 10(-6) to 10(-9) mol l(-1). Catecholamines also increased the frequency of momentary, isolated arrests of pre-oral cilia, but failed to suppress beating of the post-oral cilia at these concentrations. The neuropeptides, FMRFamide and Leu-enkephalin, did not affect the frequency of ciliary beating or of isolated ciliary arrests, but did induce numerous muscular contractions, which were accompanied by sustained ciliary arrests. In terms of whole animal behaviour, serotonin caused larvae to concentrate toward the top of a water column and to increase feeding, whereas catecholamines caused larvae to concentrate toward the bottom of a water column and decrease feeding. Monoamine analogues which facilitated or opposed the effects of synthetic transmitters on larval behaviour, further suggested that these transmitters are released endogenously to control velar function. Finally, applications of peptides to whole larvae caused increased frequency of locomotory arrests. Together these findings demonstrate several potential roles for the nervous system in controlling larval behaviour in gastropods. PMID- 17114402 TI - Photoperiod-induced plasticity of thermosensitivity and acquired thermotolerance in Locusta migratoria. AB - The mechanisms by which different life histories affect neural circuits are largely unknown. We show that the thermosensitivity and thermotolerance of neural circuit operation are affected in a complex dynamic fashion by photoperiod, prior heat experience and the sex of the animal. We compared thermosensitivity and thermotolerance of ventilatory motor pattern generation in locusts reared under two photoperiods (12:12 and 16:8; i.e. 12 h:12 h and 16 h:8 h L:D, respectively) before and after heat shock pre-treatment (HS: 3 h, 45 degrees C) in order to determine the effect of daylength on properties of neural function. We monitored central pattern generator (CPG) output electromyographically from muscle 161 in the second abdominal segment during ramped increases in temperature and also measured the time taken for the circuit to fail at high temperatures and the time taken to recover on return to room temperature. There were effects of photoperiod, heat pre-treatment and the sex of the animal on ventilatory rate, time-to-failure and time-to-recovery. The ventilatory motor pattern of 16:8 and 12:12 locusts responded differently to increasing and maintained high temperature stress in both control and heat shocked locusts. We found that 12:12 locusts were generally more robust than 16:8 locusts: they lived longer, they showed greater tolerance to high temperatures, and they recovered more quickly from temperature induced circuit failure. A faster ventilatory rate in 12:12 animals at high temperatures may have accelerated evaporative cooling to mediate improved temperature tolerance. PMID- 17114403 TI - A critical analysis of carbonic anhydrase function, respiratory gas exchange, and the acid-base control of secretion in the rectal gland of Squalus acanthias. AB - We compared in vivo responses of rectal gland secretion to carbonic anhydrase (CA) inhibition (10(-4) mol l(-1) acetazolamide) in volume-loaded dogfish with in vitro responses in an isolated-perfused gland stimulated with 5 x 10(-6) mol l( 1) forskolin and removed from systemic influences. We also measured respiratory gas exchange in the perfused gland, described the acid-base status of the secreted fluid, and determined the relative importance of various extracellular and intracellular acid-base parameters in controlling rectal gland secretion in vitro. In vivo, acetazolamide inhibited Cl(-) secretion and decreased pHi in the rectal gland, but interpretation was confounded by an accompanying systemic respiratory acidosis, which would also have contributed to the inhibition. In the perfused gland, M(CO(2)) and M(O(2)) increased in linear relation to increases in Cl(-) secretion rate. CA inhibition (10(-4) mol l(-1) acetazolamide) had no effect on Cl(-) secretion rate or pHi in the perfused gland, in contrast to in vivo, but caused a transitory 30% inhibition of M(CO(2)) (relative to stable M(O(2))) and elevation in secretion P(CO(2)) effects, which peaked at 2 h and attenuated by 3.5-4 h. Secretion was inhibited by acidosis and stimulated by alkalosis; the relationship between relative Cl(-) secretion rate and pHe was almost identical to that seen in vivo. Experimental manipulations of perfusate pH, P(CO(2)) and HCO(3)(-) concentration, together with measurements of pHi, demonstrated that these responses were most strongly correlated with changes in pHe, and were not related to changes in P(CO(2)), extracellular HCO(3)(-), or intracellular HCO(3)(-) levels, though changes in pHi may also have played a role. The acid-base status of the secreted fluid varied with that of the perfusate, secretion pH remaining about 0.3-0.5 units lower, and changing in concert with pHe rather than pHi; secretion HCO(3)(-) concentrations remained low, even in the face of greatly elevated perfusate HCO(3)(-) concentrations. We conclude that pH effects on rectal gland secretion rate are adaptive, that CA functions to catalyze the hydration of CO(2), thereby maintaining a gradient for diffusive efflux of CO(2) from the working cells, and that differences in response to CA inhibition likely reflect the higher perfusion-to-secretion ratio in vitro than in vivo. PMID- 17114404 TI - Cuttlefish responses to visual orientation of substrates, water flow and a model of motion camouflage. AB - Low-level mechanisms in vertebrate vision are sensitive to line orientation. Here we investigate orientation sensitivity in the cuttlefish Sepia pharaonis, by allowing animals to settle on stripe patterns. When camouflaging themselves cuttlefish are known to be sensitive to image parameters such as contrast and spatial scale, but we find no effect of background orientation on the patterns displayed. It is nonetheless clear that the animals see orientation, because they prefer to rest with the body-axis perpendicular to the stripes. We consider three possible mechanisms to account for this behaviour. Firstly, that the body patterns are themselves oriented, and that the cuttlefish align themselves to aid static camouflage. This is unlikely, as the patterns displayed have no dominant orientation at any spatial scale. A second possibility is that motion camouflage favours alignment of the body orthogonal to background stripes, and we suggest how this alignment can minimise motion signals produced by occlusion. Thirdly we show that cuttlefish prefer to rest with their body-axis parallel to the water flow, and it is possible that they use visual patterns such as sand ripples to determine water flow. PMID- 17114405 TI - Ontogeny of swim performance and mechanics in bottlenose dolphins (Tursiops truncatus). AB - Morphological and physiological development impacts swimming performance throughout ontogeny. Our investigation of the ontogeny of swim performance (mean and maximum swim speed) and swim effort (stroke amplitude and tailbeat frequency) of independently swimming bottlenose dolphins (Tursiops truncatus) demonstrated that swimming capabilities are extremely limited in calves. Mean and maximum swim speeds of 0-1-month-old calves were only 37% and 52% of that for adults, respectively, and levels similar to those of adults were not achieved until one year post-partum. Limitations in swim speed were associated with an inability to achieve mature thrusting capabilities, as stroke amplitude and distance covered per stroke remained significantly lower than adult levels during the first-year post-partum. Although calves were expected to require less thrusting power to propel their smaller bodies through water, size-specific stroke amplitudes of 0-3 month-olds (23-26% of body length) were smaller than those of dolphins >or=10 months post-partum (29-30% of body length). As a result, swim speed standardized by body length was significantly slower for 0-3-month-old dolphins compared with dolphins >or=10 months post-partum. These results suggest that other factors, such as underdeveloped physiology, act synergistically with small body size to limit independent swim performance in dolphins during ontogeny. PMID- 17114406 TI - Biomimetic evolutionary analysis: testing the adaptive value of vertebrate tail stiffness in autonomous swimming robots. AB - For early vertebrates, a long-standing hypothesis is that vertebrae evolved as a locomotor adaptation, stiffening the body axis and enhancing swimming performance. While supported by biomechanical data, this hypothesis has not been tested using an evolutionary approach. We did so by extending biomimetic evolutionary analysis (BEA), which builds physical simulations of extinct systems, to include use of autonomous robots as proxies of early vertebrates competing in a forage navigation task. Modeled after free-swimming larvae of sea squirts (Chordata, Urochordata), three robotic tadpoles (;Tadros'), each with a propulsive tail bearing a biomimetic notochord of variable spring stiffness, k (N m(-1)), searched for, oriented to, and orbited in two dimensions around a light source. Within each of ten generations, we selected for increased swimming speed, U (m s(-1)) and decreased time to the light source, t (s), average distance from the source, R (m) and wobble maneuvering, W (rad s(-2)). In software simulation, we coded two quantitative trait loci (QTL) that determine k: bending modulus, E (Nm(-2)) and length, L (m). Both QTL were mutated during replication, independently assorted during meiosis and, as haploid gametes, entered into the gene pool in proportion to parental fitness. After random mating created three new diploid genotypes, we fabricated three new offspring tails. In the presence of both selection and chance events (mutation, genetic drift), the phenotypic means of this small population evolved. The classic hypothesis was supported in that k was positively correlated (r(2)=0.40) with navigational prowess, NP, the dimensionless ratio of U to the product of R, t and W. However, the plausible adaptive scenario, even in this simplified system, is more complex, since the remaining variance in NP was correlated with the residuals of R and U taken with respect to k, suggesting that changes in k alone are insufficient to explain the evolution of NP. PMID- 17114407 TI - Magnetic compass in the cornea: local anaesthesia impairs orientation in a mammal. AB - The mechanism of signal transduction during magnetic compass orientation is rarely evident in vertebrates and is as yet unknown in mammals. This transmission has been associated with magnetite-based receptors innervated by the ophthalmic nerve or with the involvement of the eye, particularly the retina. We provide the first behavioural support for the cornea carrying the respective primary sensors in mole-rats (Fukomys anselli) by showing that local anaesthesia disrupts their normal directional magnetic orientation. During corneal anaesthesia in normal geomagnetic conditions, mole-rats did not maintain their preferred nesting direction, but displayed a random orientation pattern. A second experiment showed that the ability of the mole-rat to discriminate between light and dark was not impeded by the same anaesthetic treatment, suggesting no retinal involvement in mole-rat magnetic orientation. Our study restricts the peripheral primary sensors in mole-rats to the ophthalmic region, probably the cornea and indicates magnetite as the responsible signal mediator. PMID- 17114408 TI - Regulation of troponin T expression during muscle development in sea bream Sparus auratus Linnaeus: the potential role of thyroid hormones. AB - In the sea bream Sparus auratus three stage-specific fast troponin T (fTnT) isoforms have been cloned and correspond to embryonic-, larval- and adult specific isoforms. Characterisation, using database searches, of the putative genomic organisation of Fugu rubripes and Tetraodon nigroviridis fTnT indicates that alternative exon splicing in the 5 region of the gene generates the different isoforms. Moreover, comparison of teleost fTnTs suggests that alternative splicing of fTnT appears to be common in teleosts. A different temporal expression pattern for each fTnT splice varotnt is found during sea bream development and probably relates to differing functional demands, as a highly acidic embryonic form (pI 5.16) is substituted by a basic larval form (pI 9.57). Thyroid hormones (THs), which play an important regulatory role in muscle development in flatfish and tetrapods, appear also to influence TnT gene expression in the sea bream. However, THs have a divergent action on different sea bream TnT genes and although the slow isoform (sTnT1) is TH-responsive, fTnT, sTnT2 and the itronless isoform (iTnT) are unaffected. The present results taken together with those published for flatfish seem to suggest differences may exist in the regulation of larval muscle development in teleosts. PMID- 17114409 TI - Path selection in cockroaches. AB - In gregarious insects, the exploration and the use of the home range can involve both individual navigational abilities and/or chemical trails. Trail formation can result from an active laying of pheromones but can also derive from the incidental deposition of chemical cues. In this study, we investigated whether scent trails can influence path selection in the cockroach Blattella germanica (L.). Experiments were designed to separate the role of prior experience based on the orientation of the path and the presence of trails. In a first phase, cockroaches were able to access freely one or two branches of a platform during a 48 h period. In a second phase, cockroaches were offered a binary choice between one marked and one clean branch, or between two clean branches. In the absence of trails, cockroaches prefer the novel orientation but in the presence of a trail previously laid by the same group of individuals, they prefer the path with the trail, irrespective of orientation. However, cockroaches tended to avoid trails laid by a different group of cockroaches. Overall, our results indicate that both scent cues and response to novelty influence, weakly but significantly, path selection in cockroaches. The plausible nature of scent marks used by cockroaches is discussed. Our study suggests that the influence of incident trailing cues can be modulated by learning to support a flexible orientation strategy depending on individual experience. PMID- 17114410 TI - Cone photoreceptor oil droplet pigmentation is affected by ambient light intensity. AB - The cone photoreceptors of many vertebrates contain spherical organelles called oil droplets. In birds, turtles, lizards and some lungfish the oil droplets are heavily pigmented and function to filter the spectrum of light incident upon the visual pigment within the outer segment. Pigmented oil droplets are beneficial for colour discrimination in bright light, but at lower light levels the reduction in sensitivity caused by the pigmentation increasingly outweighs the benefits generated by spectral tuning. Consequently, it is expected that species with pigmented oil droplets should modulate the density of pigment in response to ambient light intensity and thereby regulate the amount of light transmitted to the outer segment. In this study, microspectrophotometry was used to measure the absorption spectra of cone oil droplets in chickens (Gallus gallus domesticus) reared under bright (unfiltered) or dim (filtered) sunlight. Oil droplet pigmentation was found to be dependent on the intensity of the ambient light and the duration of exposure to the different lighting treatments. In adult chickens reared in bright light, the oil droplets of all cone types (except the violet sensitive single cones, whose oil droplet is always non-pigmented) were more densely pigmented than those in chickens reared in dim light. Calculations show that the reduced levels of oil droplet pigmentation in chickens reared in dim light would increase the sensitivity and spectral bandwidth of the outer segment significantly. The density of pigmentation in the oil droplets presumably represents a trade-off between the need for good colour discrimination and absolute sensitivity. This might also explain why nocturnal animals, or those that underwent a nocturnal phase during their evolution, have evolved oil droplets with low pigment densities or no pigmentation or have lost their oil droplets altogether. PMID- 17114411 TI - Correlation of C-start behaviors with neural activity recorded from the hindbrain in free-swimming goldfish (Carassius auratus). AB - Startle behaviors in teleost fishes are well suited for investigations of mechanisms of sensorimotor integration because the behavior is quantifiable and much of the underlying circuitry has been identified. The teleost C-start is triggered by an action potential in one of the two Mauthner (M) cells. To correlate C-start behavior with electrophysiology, extracellular recordings were obtained from the surface of the medulla oblongata in the hindbrain, close to the M-axons, in freely swimming goldfish monitored using high-speed video. The recordings included action potentials generated by the two M-axons, as well as neighboring axons in the dorsal medial longitudinal fasciculus. Axonal backfills indicated that the latter originate from identifiable reticulospinal somata in rhombomeres 2-8 and local interneurons. Diverse auditory and visual stimuli evoked behaviors with kinematics characteristic of the C-start, and the amplitude of the first component of the hindbrain field potential correlated with the C start direction. The onset of the field potential preceded that of the simultaneously recorded trunk EMG and movement initiation by 1.08+/-0.04 and 8.13+/-0.17 ms, respectively. A subsequent longer latency field potential was predictive of a counterturn. These results indicate that characteristic features of the C-start can be extracted from the neural activity of the M-cell and a population of other reticulospinal neurons in free-swimming goldfish. PMID- 17114412 TI - Lack of generalization of object discrimination between spatial contexts by a bat. AB - Discrimination and generalization are important elements of cognition in the daily lives of animals. Nectar-feeding bats detect flowers by olfaction and probably vision, but also use echolocation and echo-perception of flowers in immediate target surroundings. The echo received from an interference-rich flower corolla is a function of a bat's own relative position in space. This raises the question how easily a free-flying bat will generalize an echo stimulus from a learning situation to a new spatial context where differences in relative flight approach trajectories may lead to an unfamiliar spectral composition of the self generated echoes. We trained free-flying Glossophaga soricina in echoacoustic discrimination in a two-alternative forced-choice (2-AFC) paradigm at location A. We then tested at location B for spontaneous transfer of discrimination ability. Bats did not spontaneously transfer the discrimination ability acquired at A to location B. This lack of spontaneous generalization may have been caused by factors of the underlying learning mechanisms. 2-AFC tasks may not be representative of the natural foraging behaviour of flower-visiting bats. In contrast to insect-eating bats that constantly evaluate the environment to detect unpredictable prey, the spatial stability of flowers may allow flower visitors to rely on spatial memory to guide foraging. The 2-AFC task requires the disregard (learned irrelevance) of salient spatial location cues that are different at each new location. In Glossophaga, a conjunction between spatial context and 2-AFC discrimination learning may have inhibited the transfer of learned irrelevance of spatial location in the 2-AFC task to new spatial locations. Alternatively, the bats may have learnt the second discrimination task completely anew, and were faster only because of an acquired learning set. We suggest a dissociation between 2-AFC task acquisition and novel object discrimination learning to resolve the issue. PMID- 17114413 TI - Structure and sexual dimorphism of the electrocommunication signals of the weakly electric fish, Adontosternarchus devenanzii. AB - Electrocommunication signals of electric fish vary across species, sexes and individuals. The diversity of these signals and the relative simplicity of the neural circuits controlling them make them a model well-suited for studying the mechanisms, evolution and sexual differentiation of behavior. In most wave-type gymnotiform knifefishes, electric organ discharge (EOD) frequency and EOD modulations known as chirps are sexually dimorphic. In the most speciose gymnotiform family, the Apteronotidae, EOD frequency is higher in males than females in some species, but lower in males than females in others. Sex differences in EOD frequency and chirping, however, have been examined in only three apteronotid species in a single genus, Apteronotus. To understand the diversity of electrocommunication signals, we characterized these behaviors in another genus, Adontosternarchus. Electrocommunication signals of Adontosternarchus devenanzii differed from those of Apteronotus in several ways. Unlike in Apteronotus, EOD frequency was not sexually dimorphic in A. devenanzii. Furthermore, although A. devenanzii chirped in response to playbacks simulating conspecific EODs, the number of chirps did not vary with different stimulus frequencies. A. devenanzii chirps also differed in structure from Apteronotus chirps. Whereas Apteronotus species produce functionally distinct chirp types differing in frequency modulation (FM), A. devenanzii produced only high frequency chirps that had either single or multiple frequency peaks. Males produced more multi-peaked chirps than females. Thus, the temporal structure of chirps, rather than the amount of FM, delineated chirp types in A. devenanzii. Our results demonstrate that the structure, function and sexual dimorphism of electrocommunication signals are evolutionary labile in apteronotids and may be useful for understanding the diversity of sexually dimorphic behavior. PMID- 17114414 TI - A nuclear factor in B cells and beyond. PMID- 17114415 TI - Multiple nuclear factors interact with the immunoglobulin enhancer sequences. Cell 1986. 46: 705-716. PMID- 17114416 TI - DUSP meet immunology: dual specificity MAPK phosphatases in control of the inflammatory response. AB - The MAPK family members p38, JNK, and ERK are all activated downstream of innate immunity's TLR to induce the production of cytokines and inflammatory mediators. However, the relative intensity and duration of the activation of different MAPK appears to determine the type of immune response. The mammalian genome encodes a large number of dual specificity phosphatases (DUSP), many of which act as MAPK phosphatases. In this study, we review the emergence of several DUSP as genes that are differentially expressed and regulated in immune cells. Recently, a series of investigations in mice deficient in DUSP1, DUSP2, or DUSP10 revealed specificity in the regulation of the different MAPK proteins, and defined essential roles in models of local and systemic inflammation. The DUSP family is proposed as a set of molecular control devices specifying and modulating MAPK signaling, which may be targeted to unleash or attenuate innate and adaptive immune effector functions. PMID- 17114417 TI - Cutting Edge: TLR3 stimulation suppresses experimental autoimmune encephalomyelitis by inducing endogenous IFN-beta. AB - Experimental autoimmune encephalomyelitis is a well-characterized model of cell mediated autoimmunity. TLRs expressed on APCs recognize microbial components and induce innate immune responses, leading to the elimination of invading infectious agents. Certain TLR agonists have been reported to have adjuvant properties in CNS autoimmune inflammatory demyelination. We report in this study that TLR3 stimulation by polyinosinic-polycytidylic acid, a double-stranded RNA analog, suppresses relapsing demyelination in a murine experimental autoimmune encephalomyelitis model. Disease suppression is associated with the induction of endogenous IFN-beta and the peripheral induction of the CC chemokine CCL2. These data indicate that a preferential activation of the MyD88-independent, type I IFN inducing TLR pathway has immunoregulatory potential in this organ-specific autoimmune disease. PMID- 17114418 TI - Cutting Edge: Plasmacytoid dendritic cells provide innate immune protection against mucosal viral infection in situ. AB - Dendritic cells (DCs) are powerful APCs capable of activating naive lymphocytes. Of the DC subfamilies, plasmacytoid DCs (pDCs) are unique in that they secrete high levels of type I IFNs in response to viruses but their role in inducing adaptive immunity remains divisive. In this study, we examined the importance of pDCs and their ability to recognize a virus through TLR9 in immunity against genital HSV-2 infection. We show that a low number of pDCs survey the vaginal mucosa at steady state. Upon infection, pDCs are recruited to the vagina and produce large amounts of type I IFNs in a TLR9-dependent manner and suppress local viral replication. Although pDCs are critical in innate defense against genital herpes challenge, adaptive Th1 immunity developed normally in the absence of pDCs. Thus, by way of migrating directly into the peripheral mucosa, pDCs act strictly as innate antiviral effector cells against mucosal viral infection in situ. PMID- 17114419 TI - Cutting Edge: IL-12 inversely regulates T-bet and eomesodermin expression during pathogen-induced CD8+ T cell differentiation. AB - Cytokines are critical determinants for specification of lineage-defining transcription factors of CD4+ T cell subsets. Little is known, however, about how cytokines regulate expression of T-bet and eomesodermin (Eomes) in effector and memory CD8+ T cells. We now report that IL-12, a signature of cell-mediated immunity, represses Eomes while positively regulating T-bet in effector CD8+ T cells during infection with Listeria monocytogenes. After resolution of infection and abatement of IL-12 signaling, Eomes expression rises whereas T-bet expression declines in memory CD8+ T cells. Eomes becomes derepressed in effector cells by ablation of IL-12 signaling. In the absence of IL-12, the dynamics of clonal expansion and contraction are also perturbed. Together, these results reveal how a pathogen-associated signal, such as IL-12, could act as a switch, regulating appropriate clonal growth and decline while, in parallel, shaping a unique pattern of fate-determining transcription factors. PMID- 17114420 TI - Cutting Edge: Pivotal function of Ubc13 in thymocyte TCR signaling. AB - The Ubc13 E2 ubiquitin-conjugating enzyme is essential for BCR-, TLR-, and IL-1 receptor (IL-1R)-mediated immune responses. Although Ubc13-deficient mice show defects in BCR-, TLR/IL-1R-, or CD40-mediated activation of mitogen-activated protein kinases, the function of Ubc13 in TCR-mediated signaling and responses remains uncertain. To address this, we here generated T cell-specific conditional Ubc13-deficient mice. The frequency of T lymphocytes was severely reduced in spleens from Ubc13-deficient mice. Moreover, Ubc13-deficient thymocytes displayed defective proliferation in response to anti-CD3/CD28 or PMA/ionophore stimulation. Regarding the signal transduction, although NF-kappaB activation was modestly affected, PMA/ionophore-induced activation of Jnk and p38 was profoundly impaired in Ubc13-deficient thymocytes. In addition, PMA/ionophore-mediated ubiquitination of NF-kappaB essential modulator (NEMO)/IkappaB kinase gamma (IKKgamma) and phosphorylation of TGF-beta-activated kinase 1 (TAK1) were nearly abolished in Ubc13-deficient thymocytes. Thus, Ubc13 plays an important role in thymocyte TCR-mediated signaling and immune responses. PMID- 17114421 TI - Cutting Edge: Stimulation of dopamine D4 receptors induce T cell quiescence by up regulating Kruppel-like factor-2 expression through inhibition of ERK1/ERK2 phosphorylation. AB - The neurotransmitter dopamine (DA) is an important regulator of human T cell functions. Although it has been observed that DA, by acting through the D1/D5, D2, and D3 receptors, can activate resting T cells by stimulating the release of cytokines and the expression of surface integrins and also inhibit the proliferation of activated T cells by down-regulating nonreceptor tyrosine kinases, there is not yet a report indicating the functional significance of the D4 DA receptors present in these cells. The present work, for the first time, demonstrates that the stimulation of D4 DA receptors in human T cells induces T cell quiescence by up-regulating lung Kruppel-like factor-2 expression through the inhibition of ERK1/ERK2 phosphorylation. These results reveal a new link between the nervous system and T cell quiescence and indicate that D4 DA receptor agonists may have a therapeutic value in diseases with uncontrolled T cell proliferation. PMID- 17114422 TI - CCR4 is a key modulator of innate immune responses. AB - CCR4 is recognized as a key receptor in Th2-associated immune processes, although very little is known about its role in innate immunity. Previous studies reported increased resistance to LPS-induced lethality in CCR4(-/-) mice compared with wild-type mice. This study demonstrates that CCR4(-/-) mice are similarly resistant to challenge with other TLR agonists, as well as bacterial peritonitis. Resistance was associated with enhanced early leukocyte recruitment, increased TLR expression, a skewed type 2 cytokine/chemokine profile, and improved bacterial clearance. Macrophages from CCR4(-/-) mice exhibited many features consistent with alternative activation, including elevated secretion of type 2 cytokines/chemokines and the found in inflammatory zone 1 (FIZZ1) protein. MyD88 dependent NF-kappaB signaling was significantly down-regulated in CCR4(-/-) macrophages, whereas p38 MAPK and JNK activation were conversely increased. These data stress the importance of CCR4 in macrophage differentiation and innate immune responses to pathogens, as well as the involvement of chemokine receptor expression in TLR signaling regulation. PMID- 17114423 TI - Cytotoxic T lymphocytes induce caspase-dependent and -independent cell death in neuroblastomas in a MHC-nonrestricted fashion. AB - The MHC class I- restricted processing and presentation pathway is frequently nonfunctional in tumor cells; therefore, the direct targeting of tumor cells by CTLs may be difficult, if at all possible, to achieve. We used neuroblastoma (NB), which represents a striking example of a tumor with an impaired MHC class I pathway, as a model to study bystander effects of activated T lymphocytes on tumor cells. We found that NB cell lines are susceptible to killing by differentiated CD8(+) CTL clones in a MHC class I-nonrestricted manner that involves two programs of cell death distinguished on the basis of different kinetics, sensitivities to caspase inhibitors, and cytokine-blocking reagents. The "early" death exhibited characteristic features of apoptosis, whereas the "delayed" caspase-independent death exhibited features associated with necrosis and was partially inhibited by TNF-alpha-blocking and prevented by overexpression of Bcl-2 or Bcl-x(L). Our data reveal a previously unappreciated complexity of death pathways induced in tumor cells by immune activation and suggest that redirecting nonspecific effector CTLs to even a small proportion of NB cells or activating CTLs in a tumor's proximity may have therapeutic effects in patients with NB. PMID- 17114424 TI - Macrophages and myeloid dendritic cells, but not plasmacytoid dendritic cells, produce IL-10 in response to MyD88- and TRIF-dependent TLR signals, and TLR independent signals. AB - We have previously reported that mouse plasmacytoid dendritic cells (DC) produce high levels of IL-12p70, whereas bone marrow-derived myeloid DC and splenic DC produce substantially lower levels of this cytokine when activated with the TLR-9 ligand CpG. We now show that in response to CpG stimulation, high levels of IL-10 are secreted by macrophages, intermediate levels by myeloid DC, but no detectable IL-10 is secreted by plasmacytoid DC. MyD88-dependent TLR signals (TLR4, 7, 9 ligation), Toll/IL-1 receptor domain-containing adaptor-dependent TLR signals (TLR3, 4 ligation) as well as non-TLR signals (CD40 ligation) induced macrophages and myeloid DC to produce IL-10 in addition to proinflammatory cytokines. IL 12p70 expression in response to CpG was suppressed by endogenous IL-10 in macrophages, in myeloid DC, and to an even greater extent in splenic CD8alpha(-) and CD8alpha(+) DC. Although plasmacytoid DC did not produce IL-10 upon stimulation, addition of this cytokine exogenously suppressed their production of IL-12, TNF, and IFN-alpha, showing trans but not autocrine regulation of these cytokines by IL-10 in plasmacytoid DC. PMID- 17114425 TI - A novel E3 ubiquitin ligase substrate screen identifies Rho guanine dissociation inhibitor as a substrate of gene related to anergy in lymphocytes. AB - Ubiquitination of eukaryotic proteins regulates a broad range of cellular processes, including regulation of T cell activation and tolerance. We have previously demonstrated that gene related to anergy in lymphocytes (GRAIL), a ring finger ubiquitin E3 ligase, is required for the induction of T cell anergy; however, the substrate(s) for GRAIL E3 ligase activity is/are unknown. In this study, we report a novel prokaryotic system developed to screen for substrates of E3 ligases. Using this screen, Rho guanine dissociation inhibitor (RhoGDI) was identified as a potential substrate of GRAIL. GRAIL was subsequently demonstrated to bind and ubiquitinate RhoGDI, although GRAIL-mediated ubiquitination of RhoGDI did not result in proteosomal degradation. Expression of GRAIL in T cells resulted in specific inhibition of RhoA GTPase activation; activation of Rac1, cdc42, and Ras GTPases were not affected. Interestingly, stable T cell lines expressing dominant-negative RhoA mimicked the GRAIL-mediated IL-2 inhibition phenotype, and T cells expressing constitutively active RhoA were able to overcome GRAIL-mediated inhibition of IL-2 expression. These findings validate our prokaryotic screen as a method of identifying substrates for ubiquitin E3 ligases and suggest a role for Rho effector molecules in T cell anergy. PMID- 17114426 TI - Paternal antigen-bearing cells transferred during insemination do not stimulate anti-paternal CD8+ T cells: role of estradiol in locally inhibiting CD8+ T cell responses. AB - Maternal immunological tolerance of the semiallogeneic fetus involves several overlapping mechanisms to balance maternal immunity and fetal development. Anti paternal CD8+ T cells are suppressed during pregnancy in some but not all mouse models. Since semen has been shown to mediate immune modulation, we tested whether exposure to paternal Ag during insemination activated or tolerized anti paternal CD8+ T cells. The uterine lumen of mated female mice contained male MHC I+ cells that stimulated effector, but not naive, CD8+ T cells ex vivo. Maternal MHC class I+ myeloid cells fluxed into the uterine lumen in response to mating and cross-presented male H-Y Ag to effector, but not naive, CD8+ T cells ex vivo. However, neither unprimed nor previously primed TCR-transgenic CD8+ T cells specific for either paternal MHC I or H-Y Ag proliferated in vivo after mating. These T cells subsequently responded normally to i.p. challenge, implicating ignorance rather than anergy as the main reason for the lack of response. CD8+ T cells responded to either peptide Ag or male cells delivered intravaginally in ovariectomized mice, but this response was inhibited by systemic estradiol (inducing an estrus-like state). Subcutaneous Ag induced responses in both cases. Allogeneic dendritic cells did not induce responses intravaginally even in ovariectomized mice in the absence of estradiol. These results suggest that inhibition of antiallogeneic responses is restricted both locally to the reproductive tract and temporally to the estrous phase of the menstrual cycle, potentially decreasing the risk of maternal immunization against paternal Ags during insemination. PMID- 17114427 TI - IL-27 induces Th1 differentiation via p38 MAPK/T-bet- and intercellular adhesion molecule-1/LFA-1/ERK1/2-dependent pathways. AB - IL-27, a novel member of the IL-6/IL-12 family, activates both STAT1 and STAT3 through its receptor, which consists of WSX-1 and gp130 subunits, resulting in positive and negative regulations of immune responses. We recently demonstrated that IL-27 induces Th1 differentiation through ICAM-1/LFA-1 interaction in a STAT1-dependent, but T-bet-independent mechanism. In this study, we further investigated the molecular mechanisms by focusing on p38 MAPK and ERK1/2. IL-27 induced Th1 differentiation was partially inhibited by lack of T-bet expression or by blocking ICAM-1/LFA-1 interaction with anti-ICAM-1 and/or anti-LFA-1, and further inhibited by both. Similarly, the p38 MAPK inhibitor, SB203580, or the inhibitor of ERK1/2 phosphorylation, PD98059, partially suppressed IL-27-induced Th1 differentiation and the combined treatment completely suppressed it. p38 MAPK was then revealed to be located upstream of T-bet, and SB203580, but not PD98059, inhibited T-bet-dependent Th1 differentiation. In contrast, ERK1/2 was shown to be located downstream of ICAM-1/LFA-1, and PD98059, but not SB203580, inhibited ICAM-1/LFA-1-dependent Th1 differentiation. Furthermore, it was demonstrated that STAT1 is important for IL-27-induced activation of ERK1/2, but not p38 MAPK, and that IL-27 directly induces mRNA expression of growth arrest and DNA damage inducible 45gamma, which is known to mediate activation of p38 MAPK. Finally, IL 12Rbeta2 expression was shown to be up-regulated by IL-27 in both T-bet- and ICAM 1/LFA-1-dependent mechanisms. Taken together, these results suggest that IL-27 induces Th1 differentiation via two distinct pathways, p38 MAPK/T-bet- and ICAM 1/LFA-1/ERK1/2-dependent pathways. This is in contrast to IL-12, which induces it via only p38 MAPK/T-bet-dependent pathway. PMID- 17114428 TI - Loss of invariant chain protects nonobese diabetic mice against type 1 diabetes. AB - The invariant (Ii) chain acts as an essential chaperone to promote MHC class II surface expression, Ag presentation, and selection of CD4(+) T cells. We have examined its role in the development of type 1 diabetes in NOD mice and show that Ii chain-deficient NOD mice fail to develop type 1 diabetes. Surprisingly, Ii chain functional loss fails to disrupt in vitro presentation of islet Ags, in the context of NOD I-A(g7) molecules. Moreover, pathogenic effector cells could be shown to be present in Ii chain-deficient NOD mice because they were able to transfer diabetes to NOD.scid recipients. The ability of these cells to transfer diabetes was markedly enhanced by depletion of CD25 cells coupled with in vivo anti-CD25 treatment of recipient mice. The numbers of CD4(+)CD25(+)Foxp3(+) T cells in thymus and periphery of Ii chain-deficient NOD mice were similar to those found in normal NOD mice, in contrast to conventional CD4(+) T cells whose numbers were reduced. This suggests that regulatory T cells are unaffected in their selection and survival by the absence of Ii chain and that an alteration in the balance of effector to regulatory T cells contributes to diabetes prevention. PMID- 17114429 TI - The chemokine CX3CL1 reduces migration and increases adhesion of neurons with mechanisms dependent on the beta1 integrin subunit. AB - Fractalkine/CX3CL1 and its specific receptor CX3CR1 are constitutively expressed in several regions of the CNS and are reported to mediate neuron-microglial interaction, synaptic transmission, and neuronal protection from toxic insults. CX3CL1 is released both by neuronal and astrocytic cells, whereas CX3CR1 is mainly expressed by microglial cells and neurons. Microglial cells efficiently migrate in response to CX3CL1, whereas no evidence is reported to date on CX3CL1 induced neuronal migration. For this reason, we have investigated in vitro the effects of CX3CL1 on basal migration of neurons and of the microglial and astrocytic populations, all these cells being obtained from the hippocampus and the cerebellum of newborn rats. We report that CX3CL1 stimulates microglial cell migration but efficiently reduces basal neuronal movement, regardless of the brain source. The effect of CX3CL1 is pertussis toxin (PTX) sensitive and PI3K dependent on hippocampal neurons, while it is PTX sensitive, PI3K dependent, and ERK dependent on cerebellar granules. Interestingly, CX3CL1 also increases neuron adhesion to the extracellular matrix component laminin, with mechanisms dependent on PTX-sensitive G proteins, and on the ERK and PI3K pathways. Both the reduction of migration and the increase of neuron adhesion require the activation of the beta(1) and alpha(6) integrin subunits with the exception of cerebellar neuron migration, which is only dependent on the beta(1) subunit. More importantly, in neurons, CX3CL1/CXCL12 cotreatment abolished the effect mediated by a single chemokine on chemotaxis and adhesion. In conclusion, our findings indicate that CX3CL1 reduces neuronal migration by increasing cell adhesion through integrin dependent mechanisms in hippocampal and cerebellar neurons. PMID- 17114430 TI - ERK5 activates NF-kappaB in leukemic T cells and is essential for their growth in vivo. AB - MAPK cascades play a central role in the cellular response to the environment. The pathway involving the MAPK ERK5 mediates growth factor- and stress-induced intracellular signaling that controls proliferation or survival depending upon the cell context. In this study, we show that reducing ERK5 levels with a specific small hairpin RNA 5 (shERK5) reduced cell viability, sensitized cells to death receptor-induced apoptosis, and blocked the palliative effects of phorbol ester in anti-Fas Ab-treated cells. shERK5 decreased nuclear accumulation of the NF-kappaB p65 subunit, and conversely, ectopic activation of ERK5 led to constitutive nuclear localization of p65 and increased its ability to trans activate specific reporter genes. Finally, the T lymphoma cell line EL-4, upon expression of shERK5, proliferated in vitro, but failed to induce s.c. tumors in mice. Our results suggest that ERK5 is essential for survival of leukemic T cells in vivo, and thus represents a promising target for therapeutic intervention in this type of malignancy. PMID- 17114431 TI - IL-12-programmed long-term CD8+ T cell responses require STAT4. AB - Immunological adjuvants activate innate immune cells for Ag presentation and elicitation of cytokines like IL-12 that promote T cell expansion and effector differentiation. An important but elusive aim for most immunization strategies is to produce memory T cells that provide durable immunity. Recent evidence demonstrates that the context of Ag presentation instructionally programs T cells for short- and long-term responses. However, the role and mechanisms by which cytokines like IL-12 condition CD8 T cells for long-term responses remain relatively uncharacterized. In this study, we show that brief exposure (20 h) of naive TCR-transgenic CD8 cells to IL-12 during Ag stimulation leads to transient phosphorylation of STAT4 for robust effector differentiation. Moreover, the IL-12 induced STAT4 engenders greater clonal expansion of the Ag-activated CD8 cells by regulating the expression of the transcriptional factor Bcl3- and Bcl2-related genes that promote survival of Ag-activated CD8 cells. Remarkably, the IL-12 conditioned CD8 T cells demonstrate increased sensitivity to IL-7 and IL-15, whereby they are rendered "fit" for homeostatic self-renewal as well as augmented CD4-dependent recall responses that are effective at controlling Salmonella infection in vivo. This information provides new insights into mechanisms by which IL-12 conditions CD8 T cells for long-term immunity, which is likely to benefit development of new strategies for the use of IL-12 in infectious diseases and cancer. PMID- 17114432 TI - Distinct and non-overlapping T cell receptor repertoires expanded by DNA vaccination in wild-type and HER-2 transgenic BALB/c mice. AB - Central tolerance to tumor-associated Ags is an immune-escape mechanism that significantly limits the TCR repertoires available for tumor eradication. The repertoires expanded in wild-type BALB/c and rat-HER-2/neu (rHER-2) transgenic BALB-neuT mice following DNA immunization against rHER-2 were compared by spectratyping the variable (V)beta and the joining (J)beta CDR 3. Following immunization, BALB/c mice raised a strong response. Every mouse used one or more CD8+ T cell rearrangements of the Vbeta9-Jbeta1.2 segments characterized by distinct length of the CDR3 and specific for 63-71 or 1206-1214 rHER-2 peptides. In addition, two CD4+ T cell rearrangements recurred in >50% of mice. Instead, BALB-neuT mice displayed a limited response to rHER-2. Their repertoire is smaller and uses different rearrangements confined to CD4+ T cells. Thus, central tolerance in BALB-neuT mice acts by silencing the BALB/c mice self-reactive repertoire and reducing the size of the CD8+ T cell component. CD8+ and CD4+ T cells from both wild-type and transgenic mice home to tumors. This definition of the T cell repertoires available is critical to the designing of immunological maneuvers able to elicit an effective immune reaction against HER-2-driven carcinogenesis. PMID- 17114433 TI - Oral tolerance induction with antigen conjugated to cholera toxin B subunit generates both Foxp3+CD25+ and Foxp3-CD25- CD4+ regulatory T cells. AB - Oral administration of Ag coupled to cholera toxin B subunit (CTB) efficiently induces peripheral immunological tolerance. We investigated the extent to which this oral tolerance is mediated by CD25+CD4+ regulatory T cells (T(reg)). We found that total T(reg), KJ1-26+ T(reg) and CTLA-4+ T(reg) were all increased in Peyer's patches, mesenteric lymph nodes, and, to a lesser extent, in spleen of mice after intragastric administration of OVA/CTB conjugate, which also increased TGF-beta in serum. This could be abolished by co-administering cholera toxin or by treatment with anti-TGF-beta mAb. CD25+ T(reg), but also CD25-CD4+ T cells from OVA/CTB-treated BALB/c or DO11.10 mice efficiently suppressed effector T cell proliferation and IL-2 production in vitro. Following adoptive transfer, both T cell populations also suppressed OVA-specific T cell and delayed-type hypersensitivity responses in vivo. Foxp3 was strongly expressed by CD25+ T(reg) from OVA/CTB-treated mice, and treatment also markedly expanded CD25+Foxp3+ T(reg). Furthermore, in Rag1(-/-) mice that had adoptively received highly purified Foxp3-CD25-CD4+ OT-II T cells OVA/CTB feeding efficiently induced CD25+ T(reg) cells, which expressed Foxp3 more strongly than naturally developing T(reg) and also had stronger ability to suppress effector OT-II T cell proliferation. A remaining CD25- T cell population, which also became suppressive in response to OVA/CTB treatment, did not express Foxp3. Our results demonstrate that oral tolerance induced by CTB-conjugated Ag is associated with increase in TGF-beta and in both the frequency and suppressive capacity of Foxp3+ and CTLA-4+ CD25+ T(reg) together with the generation of both Foxp3+ and Foxp3-CD25- CD4+ T(reg). PMID- 17114434 TI - Regulation of immunity by a novel population of Qa-1-restricted CD8alphaalpha+TCRalphabeta+ T cells. AB - Regulatory mechanisms involving CD8+ T cells (CD8 regulatory T cells (Tregs)) are important in the maintenance of immune homeostasis. However, the inability to generate functional CD8 Treg clones with defined Ag specificity has precluded a direct demonstration of CD8 Treg-mediated regulation. In the present study, we describe the isolation of functional lines and clones representing a novel population of TCRalphabeta+ Tregs that control activated Vbeta8.2+ CD4 T cells mediating experimental autoimmune encephalomyelitis. They express exclusively the CD8alphaalpha homodimer and recognize a peptide from a conserved region of the TCR Vbeta8.2 chain in the context of the Qa-1a (CD8alphaalpha Tregs). They secrete type 1 cytokines but not IL-2. CD8alphaalpha Tregs kill activated Vbeta8.2+ but not Vbeta8.2- or naive T cells. The CD8alphaalpha Tregs prevent autoimmunity upon adoptive transfer or following in vivo activation. These findings reveal an important negative feedback regulatory mechanism targeting activated T cells and have implications in the development of therapeutic strategies for autoimmune diseases and transplantation. PMID- 17114435 TI - Regulation of Th2 cell development by Polycomb group gene bmi-1 through the stabilization of GATA3. AB - The Polycomb group (PcG) gene products regulate the maintenance of the homeobox gene expression in Drosophila and vertebrates and also the cell cycle progression in thymocytes and Th2 cell differentiation in mature T cells. We herein studied the role of PcG gene bmi-1 product in Th1/Th2 cell differentiation and found that Bmi-1 facilitates Th2 cell differentiation in a Ring finger-dependent manner. Biochemical studies indicate that Bmi-1 interacts with GATA3 in T cells, which is dependent on the Ring finger of Bmi-1. The overexpression of Bmi-1 resulted in a decreased ubiquitination and an increased protein stability of GATA3. In bmi-1 deficient Th cells, the levels of Th2 cell differentiation decreased as the degradation and ubiquitination on GATA3 increased. Therefore, Bmi-1 plays a crucial role in the control of Th2 cell differentiation in a Ring finger dependent manner by regulating GATA3 protein stability. PMID- 17114436 TI - Oncostatin M enhances CCL21 expression by microvascular endothelial cells and increases the efficiency of dendritic cell trafficking to lymph nodes. AB - CCL21, a lymphatic endothelial cell (LEC)-derived chemokine, and its receptor CCR7 regulate dendritic cell (DC) trafficking to lymph nodes (LN), but it is unclear how CCL21 expression is regulated. Oncostatin M (OSM) is an IL-6-like cytokine synthesized by activated DC and other leukocytes. In vitro, OSM (but not TNF-alpha) stimulated CCL21 mRNA and protein expression by human dermal microvascular EC (DMEC) in an ERK1/2-dependent fashion. Conditioned medium from OSM-treated DMEC stimulated CCL21-dependent chemotaxis of mouse bone marrow derived DC (BMDC). Cultured BMDC expressed OSM, which was increased with the addition of LPS. Topical application of the contact-sensitizing hapten, trinitrochlorobenzene, resulted in enhanced OSM expression in the skin, whereas cutaneous injection of TNF-alpha did not. Injection of OSM into the footpad increased CCL21 mRNA expression in the draining LN by approximately 10-fold and in mouse skin by approximately 4-fold without increasing CCR7 mRNA. In vitro, OSM increased the permeability of DMEC and lung microvascular EC monolayers to FITC dextran beads, and, in vivo, it enhanced accumulation of Evans blue dye in draining LN by approximately 3-fold (p = 0.0291). Of note, OSM increased trafficking of BMDC injected in footpads to draining LN by 2-fold (p = 0.016). In summary, OSM up-regulates CCL21 expression in skin and draining regional LN. We propose that OSM is a regulator of CCL21 expression and endothelial permeability in skin, contributing to efficient migration of DC to regional LN. PMID- 17114437 TI - Microenvironment-dependent requirement of STAT4 for the induction of P-selectin ligands and effector cytokines on CD4+ T cells in healthy and parasite-infected mice. AB - T effector cells require selectin ligands to migrate into inflamed regions. In vitro, IL-12 promotes induction of these ligands as well as differentiation of CD4+ T cells into IFN-gamma-producing Th1 but not Th2 cells. STAT4 is strongly involved in these processes. However, the presence of selectin ligands on various T effector cell subsets in vivo points to more complex regulatory pathways. To clarify the role of the IL-12/STAT4 signaling pathway, we analyzed the impact of STAT4 deficiency on the expression of P-selectin ligands (P-lig) on CD4+ T cells in vitro and in vivo, including conditions of infection. In vitro, we found significant expression of P-lig upon activation not only in the presence, but also in the absence, of IL-12, which was independent of STAT4. TGF-beta, an alternative inducer of selectin ligands in human T cells, was not effective in murine CD4+ T cells, suggesting a role of additional signaling pathways. In vivo, a significant impact of STAT4 for the generation of P-lig+CD4+ T cells was observed for cells from peripheral lymph nodes, but not for those from spleen or lung. However, upon infection with the Th2-inducing parasite Nippostrongylus brasiliensis, P-lig expression became dependent on STAT4 signaling. Interestingly, also the frequency of IL-4-producing cells was greatly diminished in absence of STAT4. These data reveal a hitherto unknown contribution of STAT4 to the generation of Th2 cells in parasite infection and suggest that signals inducing inflammation-seeking properties in vivo vary depending on environmental conditions, such as type of organ and infection. PMID- 17114438 TI - Immunoproteasome subunit deficiencies impact differentially on two immunodominant influenza virus-specific CD8+ T cell responses. AB - Primary CD8+ T cell (T(CD8+)) responses to viruses are directed toward multiple Ags and shaped by both the level of Ag presentation and the underlying Ag specific T(CD8+) repertoire. The relative importance of these factors in deciding the hierarchy of T(CD8+) responses and how they are influenced by the immunoproteasome are not well understood. Using an influenza infection model in mice deficient in various immunoproteasome subunits, we observe that Ag presentation and T(CD8+) repertoire are altered in an epitope-specific and immunoproteasome subunit-dependent manner. More importantly, we find that the level of Ag presentation and the extent of the underlying repertoire can work either alone or in concert to determine definitively the magnitude of the individual T(CD8+) responses and hence the overall T(CD8+) hierarchy. Together, these results provide a clearer understanding of how immunodominance hierarchies are established. PMID- 17114439 TI - Cyclosporin A abolishes CD28-mediated resistance to CD95-induced apoptosis via superinduction of caspase-3. AB - Costimulation of T cells via CD28 promotes both proliferation and resistance to apoptosis. In this study, we show that the immunosuppressive drug cyclosporin A (CsA) fully reverses resistance to CD95-mediated cell death after TCR/CD28 costimulation or superagonistic anti-CD28 mAb stimulation of primary rat lymph node T cells. This effect correlated with a pronounced superinduction of caspase 3 on both mRNA and protein levels, whereas its main antagonist, X chromosome linked inhibitor of apoptosis, was unaffected by inclusion of CsA. Apoptosis triggered by CD95 cross-linking was characterized by robust caspase-3 activation. Furthermore, CsA sensitization to CD95-mediated apoptosis of CD28-activated T cells did not alter mRNA stability of superinduced caspase-3 mRNA, suggesting a transcriptional regulation of the caspase-3 gene. Addition of Ca(2+) ionophores to TCR/CD28 or superagonistic CD28-stimulated cells reduced caspase-3 levels, further supporting a role for Ca(2+)-dependent signaling pathways in negatively regulating caspase-3. Taken together, these findings suggest that CsA promotes sensitivity to CD95-mediated apoptosis in CD28-stimulated T cells by superinduction of the caspase-3 gene via a mechanism involving suppression of the calcineurin pathway. PMID- 17114440 TI - Control of memory CD4 T cell recall by the CD28/B7 costimulatory pathway. AB - The CD28/B7 costimulatory pathway is generally considered dispensable for memory T cell responses, largely based on in vitro studies demonstrating memory T cell activation in the absence of CD28 engagement by B7 ligands. However, the susceptibility of memory CD4 T cells, including central (CD62L(high)) and effector memory (T(EM); CD62L(low)) subsets, to inhibition of CD28-derived costimulation has not been closely examined. In this study, we demonstrate that inhibition of CD28/B7 costimulation with the B7-binding fusion molecule CTLA4Ig has profound and specific effects on secondary responses mediated by memory CD4 T cells generated by priming with Ag or infection with influenza virus. In vitro, CTLA4Ig substantially inhibits IL-2, but not IFN-gamma production from heterogeneous memory CD4 T cells specific for influenza hemagglutinin or OVA in response to peptide challenge. Moreover, IL-2 production from polyclonal influenza-specific memory CD4 T cells in response to virus challenge was completely abrogated by CTLA4Ig with IFN-gamma production partially inhibited. When administered in vivo, CTLA4Ig significantly blocks Ag-driven memory CD4 T cell proliferation and expansion, without affecting early recall and activation. Importantly, CTLA4Ig treatment in vivo induced a striking shift in the phenotype of the responding population from predominantly T(EM) in control-treated mice to predominantly central memory T cells in CTLA4Ig-treated mice, suggesting biased effects of CTLA4Ig on T(EM) responses. Our results identify a novel role for CD28/B7 as a regulator of memory T cell responses, and have important clinical implications for using CTLA4Ig to abrogate the pathologic consequences of T(EM) cells in autoimmunity and chronic disease. PMID- 17114441 TI - Talin1 regulates TCR-mediated LFA-1 function. AB - The leukocyte integrin LFA-1 plays a critical role in T cell trafficking and T cell adhesion to APCs. It is known that integrin-mediated adhesion is regulated by changes in integrin ligand-binding affinity and valency through inside-out signaling. However, the molecular mechanisms involved in TCR-mediated LFA-1 regulation are not well understood. In this study, we show that the cytoskeletal protein talin1 is required for TCR-mediated activation of LFA-1 through regulation of LFA-1 affinity and clustering. Depletion of talin1 from human T cells by small interfering RNAs impairs TCR-induced adhesion to ICAM-1 and T cell APC conjugation. TCR-induced LFA-1 polarization, but not actin polarization, is defective in talin1-deficient T cells. Although LFA-1 affinity is also reduced in talin1-deficient T cells, rescue of LFA-1 affinity alone is not sufficient to restore LFA-1 adhesive function. Together, our findings indicate that TCR-induced up-regulation of LFA-1-dependent adhesiveness and resulting T cell-APC conjugation require talin1. PMID- 17114442 TI - B cell-mediated antigen presentation is required for the pathogenesis of acute cardiac allograft rejection. AB - Acute allograft rejection requires the activation of alloreactive CD4 T cells. Despite the capacity of B cells to act as potent APCs capable of activating CD4 T cells in vivo, their role in the progression of acute allograft rejection was unclear. To determine the contribution of B cell APC function in alloimmunity, we engineered mice with a targeted deficiency of MHC class II-mediated Ag presentation confined to the B cell compartment. Cardiac allograft survival was markedly prolonged in these mice as compared to control counterparts (median survival time, >70 vs 9.5 days). Mechanistically, deficient B cell-mediated Ag presentation disrupted both alloantibody production and the progression of CD4 T cell activation following heart transplantation. These findings demonstrate that indirect alloantigen presentation by recipients' B cells plays an important role in the efficient progression of acute vascularized allograft rejection. PMID- 17114443 TI - Imprinting the fate of antigen-reactive B cells through the affinity of the B cell receptor. AB - Long-lived plasma cells (PCs) and memory B cells (B(mem)) constitute the cellular components of enduring humoral immunity, whereas short-lived PCs that rapidly produce Ig correspond to the host's need for immediate protection against pathogens. In this study we show that the innate affinity of the BCR for Ag imprints upon naive B cells their differentiation fate to become short- or long lived PCs and B(mem). Using BCR transgenic mice with varying affinities for Ag, naive B cells with high affinity lose their capacity to form germinal centers (GCs), develop neither B(mem) nor long-lived PCs, and are destined to a short lived PC fate. Moderate affinity interactions result in hastened GC responses, and differentiation to long-lived PCs, but B(mem) remain extinct. In contrast, lower affinity interactions show tempered GCs, producing B(mem) and affinity matured, long-lived PCs. Thus, a continuum of elementary to comprehensive humoral immune responses exists that is controlled by inherent BCR affinity. PMID- 17114444 TI - Expression of Integrin beta3 is correlated to the properties of quiescent hemopoietic stem cells possessing the side population phenotype. AB - With significant attention paid to the field of tissue-specific stem cells, the identification of stem cell-specific markers is of considerable importance. Previously, the side population (SP) phenotype, with the capacity to efflux the DNA-binding dye Hoechst 33342, has been recognized as a common feature of adult tissue-specific stem cells. In this study, we show that high expression of integrin beta(3) (CD61) is an attribute of SP cells isolated from mouse bone marrow. Additionally, we confirmed that the expression of integrin beta(3) is correlated with properties of quiescent hemopoietic stem cells (HSCs) including the strength of the SP phenotype, cell cycle arrest, expression of HSC markers, and long-term hemopoiesis. Importantly, Lineage(-) (Lin(-))/integrin beta(3)(high) (beta(3)(high)) SP cells have as strong a capacity for long-term hemopoiesis as c-Kit(+)/Sca-1(+)/Lin(-) SP cells, which are regarded as one of the most highly enriched HSC populations. Finally, the integrin beta(3) subunit that is present in SP cells having the properties of HSCs, is associated with integrin alpha(v) (CD51). Therefore, our results demonstrate that high expression of integrin beta(3) is correlated to the properties of quiescent HSCs and suggest that the integrin beta(3) subunit is available as a common surface marker of tissue-specific stem cells. PMID- 17114445 TI - TLR2/TLR4-independent neutrophil activation and recruitment upon endocytosis of nucleosomes reveals a new pathway of innate immunity in systemic lupus erythematosus. AB - The nucleosome is a major autoantigen in systemic lupus erythematosus (SLE); it can be detected as a circulating complex in the serum, and nucleosomes have been suggested to play a key role in disease development. In the present study, we show for the first time that physiological concentrations of purified nucleosomes trigger innate immunity. The nucleosomes are endocytosed and induce the direct activation of human neutrophils (polymorphonuclear leukocytes (PMN)) as revealed by CD11b/CD66b up-regulation, IL-8 secretion, and increased phagocytic activity. IL-8 is a neutrophil chemoattractant detected in high concentrations in the sera of patients, and IL-8 secretion might thus result in enhanced inflammation, as observed in lupus patients, via an amplification loop. Nucleosomes act as free complexes requiring no immune complex formation and independently of the presence of unmethylated CpG DNA motifs. Both normal and lupus neutrophils are sensitive to nucleosome-induced activation, and activation is not due to endotoxin or high mobility group box 1 contamination. In mice, i.p. injection of purified nucleosomes induces neutrophil activation and recruitment in a TLR2/TLR4 independent manner. Importantly, neutrophils have been suggested to link innate and adaptive immunity. Thus, nucleosomes trigger a previously unknown pathway of innate immunity, which may partially explain why peripheral tolerance is broken in SLE patients. PMID- 17114446 TI - Dendritic cells amplify T cell-mediated immune responses in the central nervous system. AB - Neuroinflammation often starts with the invasion of T lymphocytes into the CNS leading to recruitment of macrophages and amplification of inflammation. In this study, we show that dendritic cells (DCs) facilitate T-T cell help in the CNS and contribute to the amplification of local neuroinflammation. We adoptively transferred defined amounts of naive TCR-transgenic (TCR) recombination activating gene-1-deficient T cells into another TCR-transgenic mouse strain expressing different Ag specificity. Following adoptive transfers, we coinjected DCs that presented one or multiple Ags into the brain and followed the activation of T cells with defined specificities simultaneously. Injection of DCs presenting both Ags simultaneously led to significantly higher infiltration of T cells into the brain compared with injection of a mixture of DCs pulsed with two Ags separately. DCs mediated either cooperative or competitive interactions between T cell populations with different specificities depending upon their MHC restricting element usage. These results suggest that DC-mediated cooperation between brain-infiltrating T cells of different Ag specificities in the CNS plays an important role in regulation of neuroinflammation. This work also implies that blocking Ag-specific responses may block not only the targeted specificities, but may also effectively block their cooperative assistance to other T cells. Therefore, these data justify more attention to Ag-specific therapeutic approaches for neuroinflammation. PMID- 17114447 TI - IL-10 inhibits lipopolysaccharide-induced CD40 gene expression through induction of suppressor of cytokine signaling-3. AB - Costimulation between T cells and APCs is required for adaptive immune responses. CD40, an important costimulatory molecule, is expressed on a variety of cell types, including macrophages and microglia. The aberrant expression of CD40 is implicated in diseases including multiple sclerosis, rheumatoid arthritis, and Alzheimer's disease, and inhibition of CD40 signaling has beneficial effects in a number of animal models of autoimmune diseases. In this study, we discovered that IL-10, a cytokine with anti-inflammatory properties, inhibits LPS-induced CD40 gene expression. We previously demonstrated that LPS induction of CD40 in macrophages/microglia involves both NF-kappaB activation and LPS-induced production of IFN-beta, which subsequently activates STAT-1alpha. IL-10 inhibits LPS-induced IFN-beta gene expression and subsequent STAT-1alpha activation, but does not affect NF-kappaB activation. Our results also demonstrate that IL-10 inhibits LPS-induced recruitment of STAT-1alpha, RNA polymerase II, and the coactivators CREB binding protein and p300 to the CD40 promoter, as well as inhibiting permissive histone H3 acetylation (AcH3). IL-10 and LPS synergize to induce suppressor of cytokine signaling (SOCS)-3 gene expression in macrophages and microglia. Ectopic expression of SOCS-3 attenuates LPS-induced STAT activation, and inhibits LPS-induced CD40 gene expression, comparable to that seen by IL-10. These results indicate that SOCS-3 plays an important role in the negative regulation of LPS-induced CD40 gene expression by IL-10. PMID- 17114448 TI - Gut IgA class switch recombination in the absence of CD40 does not occur in the lamina propria and is independent of germinal centers. AB - Conflicting findings have recently been presented as to the sites and sources of B cells that undergo class switch recombination (CSR) to IgA in the gut. In this study we provide compelling evidence in CD40(-/-) mice demonstrating that IgA CSR can be independent of CD40 signaling and germinal center formation and does not occur in the gut lamina propria (LP) itself. We found that CD40(-/-) mice had near normal levels of gut total IgA despite lacking germinal centers and completely failing to raise specific responses against the T cell-dependent Ags cholera toxin and keyhole limpet hemocyanin. The Peyer's patches in CD40(-/-) mice expressed unexpectedly high levels of activation-induced cytidine deaminase mRNA and germline alpha transcripts, but few postswitch circular DNA transcripts, arguing against significant IgA CSR. Moreover and more surprisingly, wild-type mice exhibited no to low IgA CSR in mesenteric lymph nodes or isolated lymphoid follicles. Importantly, both strains failed to demonstrate any of the molecular markers for IgA CSR in the gut LP itself. Whereas all of the classical sites for IgA CSR in the GALT in CD40(-/-) mice appeared severely compromised for IgA CSR, B cells in the peritoneal cavity demonstrated the expression of activation induced cytidine deaminase mRNA comparable to that of wild-type mice. However, peritoneal cavity B cells in both strains expressed intermediate levels of the germinal center marker GL7 and exhibited no germline alpha transcripts, and only three of 51 mice analyzed showed the presence of postswitch circular DNA transcripts. Taken together, these findings strongly argue for alternative inductive sites for gut IgA CSR against T cell-independent Ags outside of the GALT and the nonorganized LP. PMID- 17114449 TI - Characterization of myeloid and plasmacytoid dendritic cells in human lung. AB - Dendritic cells (DCs) are bone marrow-derived mononuclear cells that play a central role in the initiation of immune responses. Because human lung DCs have been incompletely characterized, we enumerated and phenotyped mononuclear cell populations from excess lung tissue obtained at surgery. Myeloid DCs (MDCs) were identified as CD1c(+)CD11c(+)CD14(-)HLA-DR(+) cells and comprised approximately 2% of low autofluorescent (LAF) mononuclear cells. Plasmacytoid DCs (PDCs) were characterized as CD123(+)CD11c(-)CD14(-)HLA-DR(+) cells and comprised approximately 1.0% of the LAF mononuclear cells. Cells enriched in MDCs expressed CD86, moderate CD80, and little CD40, but cells enriched in PDCs had little to no expression of these three costimulatory molecules. CD11c(+)CD14(-) lineage negative (MDC-enriched) LAF cells were isolated and shown to be much more potent in stimulating an alloreaction than CD11c(+)CD14(+) lineage-negative (monocyte enriched) LAF cells. PDC-enriched cells were more capable of responding to a TLR 7 agonist by secreting IFN-alpha than MDC-enriched cells. MDC-enriched cells were either CD123(+) or CD123(-), but both subsets secreted cytokines and chemokines typical of MDC upon stimulation with a TLR-4 agonist and both subsets failed to secrete IFN-alpha upon stimulation with a TLR-7 agonist. By immunohistochemistry, we identified MDCs throughout different anatomical locations of the lung. However, our method did not allow the localization of PDCs with certainty. In conclusion, in the human lung MDCs were twice as numerous and expressed higher levels of costimulatory molecules than PDCs. Our data suggest that both lung DC subsets exert distinct immune modulatory functions. PMID- 17114450 TI - Regulation of dendritic cell function and T cell priming by the fatty acid binding protein AP2. AB - The fatty acid-binding protein (FABP) family consists of a number of conserved cytoplasmic proteins with roles in intracellular lipid transport, storage, and metabolism. Examination of a comprehensive leukocyte gene expression database revealed strong expression of the adipocyte FABP aP2 in human monocyte-derived dendritic cells (DCs). We isolated bone marrow-derived DC from aP2-deficient mice, and showed that expression of DC cytokines including IL-12 and TNF was significantly impaired in these cells. Degradation of IkappaBalpha was also impaired in aP2-deficient DCs, indicative of reduced signaling through the IkappaB kinase-NF-kappaB pathway. The cytokine defect was selective because there was no effect on Ag uptake or expression of MHC class II, CD40, CD80, or CD86. In an MLR, aP2-deficient DCs stimulated markedly lower T cell proliferation and cytokine production than did wild-type DCs. Moreover, aP2-deficient mice immunized with keyhole limpet hemocyanin/CFA showed reduced production of IFN gamma by restimulated draining lymph node cells, suggesting a similar defect in DC function in vivo. Similarly, infection of aP2-deficient mice with the natural mouse pathogen ectromelia virus resulted in substantially lower production of IFN gamma by CD8+ T cells. Thus, FABP aP2 plays an important role in DC function and T cell priming, and provides an additional link between metabolic processes and the regulation of immune responses. PMID- 17114451 TI - Generation and growth of CD28nullCD8+ memory T cells mediated by IL-15 and its induced cytokines. AB - Accumulation of CD28(null)CD8+ T cells and the defects of these cells in response to antigenic stimulation are the hallmarks of age-associated decline of T cell function. However, the mechanism of these age-associated changes is not fully understood. In this study, we report an analysis of the growth of human CD28(null) and CD28+CD8+ memory T cells in response to homeostatic cytokine IL-15 in vitro. We showed that 1) there was no proliferative defect of CD28(null)CD8+ memory T cells in response to IL-15 compared with their CD28+ counterparts; 2) stable loss of CD28 expression occurred in those actively dividing CD28+CD8+ memory T cells responding to IL-15; 3) the loss of CD28 was in part mediated by TNF-alpha that was induced by IL-15; and 4) CCL4 (MIP-1beta), also induced by IL 15, had a significant inhibitory effect on the growth of CD28(null) cells, which in turn down-regulated their expression of CCL4 receptor CCR5. Together, these findings demonstrate that CD28(null)CD8+ memory T cells proliferate normally in response to IL-15 and that IL-15 and its induced cytokines regulate the generation and growth of CD28(null)CD8+ T cells, suggesting a possible role of IL 15 in the increase in CD28(null)CD8+ T cells that occurs with aging. PMID- 17114452 TI - Cyclooxygenase-2 inhibition attenuates antibody responses against human papillomavirus-like particles. AB - Vaccination to generate protective humoral immunity against infectious disease is becoming increasingly important due to emerging strains of virus, poorly immunogenic vaccines, and the threat of bioterrorism. We demonstrate that cyclooxygenase-2 (Cox-2) is crucial for optimal Ab responses to a model vaccine, human papillomavirus type 16 virus-like particles (HPV 16 VLPs). Cox-2-deficient mice produce 70% less IgG, 50% fewer Ab-secreting cells, and 10-fold less neutralizing Ab to HPV 16 VLP vaccination compared with wild-type mice. The reduction in Ab production by Cox-2(-/-) mice was partially due to a decrease in class switching. SC-58125, a structural analog of the Cox-2-selective inhibitor Celebrex reduced by approximately 70% human memory B cell differentiation to HPV 16 VLP IgG-secreting cells. The widespread use of nonsteroidal anti-inflammatory drugs and Cox-2-selective inhibitory drugs may therefore reduce vaccine efficacy, especially when vaccines are poorly immunogenic or the target population is poorly responsive to immunization. PMID- 17114453 TI - A regulatory CD4+ T cell subset in the BB rat model of autoimmune diabetes expresses neither CD25 nor Foxp3. AB - Biobreeding (BB) rats model type 1 autoimmune diabetes (T1D). BB diabetes-prone (BBDP) rats develop T1D spontaneously. BB diabetes-resistant (BBDR) rats develop T1D after immunological perturbations that include regulatory T cell (Treg) depletion plus administration of low doses of a TLR ligand, polyinosinic polycytidylic acid. Using both models, we analyzed CD4+CD25+ and CD4+CD45RC- candidate rat Treg populations. In BBDR and control Wistar Furth rats, CD25+ T cells comprised 5-8% of CD4+ T cells. In vitro, rat CD4+CD25+ T cells were hyporesponsive and suppressed T cell proliferation in the absence of TGF-beta and IL-10, suggesting that they are natural Tregs. In contrast, CD4+CD45RC(-) T cells proliferated in vitro in response to mitogen and were not suppressive. Adoptive transfer of purified CD4+CD25+ BBDR T cells to prediabetic BBDP rats prevented diabetes in 80% of recipients. Surprisingly, CD4+CD45RC-CD25- T cells were equally protective. Quantitative studies in an adoptive cotransfer model confirmed the protective capability of both cell populations, but the latter was less potent on a per cell basis. The disease-suppressing CD4+CD45RC-CD25- population expressed PD-1 but not Foxp3, which was confined to CD4+CD25+ cells. We conclude that CD4+CD25+ cells in the BBDR rat act in vitro and in vivo as natural Tregs. In addition, another population that is CD4+CD45RC-CD25- also participates in the regulation of autoimmune diabetes. PMID- 17114454 TI - Evidence for NK cell subsets based on chemokine receptor expression. AB - To help understand the role of chemokines in NK cell trafficking, we determined the chemokine receptor profiles of three different human NK cell lines and freshly isolated primary human NK cells. The cell lines overlapped in their chemokine receptor profiles: CXCR3 and CXCR4 were expressed by all three lines, whereas CCR1, CCR4, CCR6, CCR7, and CX3CR1 were expressed by only one or two of the lines, and no other chemokine receptors were detected. Freshly isolated primary NK cells were found to express CXCR1, CXCR3, and CXCR4, and to contain subsets expressing CCR1, CCR4, CCR5, CCR6, CCR7, CCR9, CXCR5, and CXCR6. With the exception of CCR4, these chemokine receptors were expressed at higher percentages by CD56(bright) NK cells than by CD56(dim) NK cells. In particular, CCR7 was expressed by almost all CD56(bright) NK cells but was not detected on CD56(dim) NK cells. CCR9 and CXCR6 have not been described previously on primary NK cells. These results indicate that within both the CD56(bright) and CD56(dim) NK cell populations, subsets with the capacity for differential trafficking programs exist, which likely influence their functions in innate and adaptive immunity. PMID- 17114455 TI - IFN-beta-mediated up-regulation of CD1d in bacteria-infected APCs. AB - The expression of CD1d molecules is essential for the selection and activation of a unique subset of T cells, invariant NKT cells, which express limited TCR diversity and have been demonstrated to function in both regulatory and antimicrobial immune responses. Although it has been reported that the levels of CD1d expression can be modulated during infection, the mechanisms that mediate this effect are poorly defined. In this study, we show that infection of dendritic cells and macrophages both in vitro and in vivo with the intracellular pathogen Listeria monocytogenes leads to up-regulation of CD1d. IFN-beta is required to mediate this up-regulation in L. monocytogenes infection, as well as being sufficient to up-regulate CD1d expression in vitro. Unlike MHC class I molecules, the increased surface expression of CD1d by IFN-beta is not regulated at the transcriptional level. Confocal microscopy and metabolic labeling experiments show that the total pool of CD1d protein is increased in IFN-beta treated cells and that increased surface expression of CD1d is not due to the redistribution of the intracellular pool of CD1d. IFN-beta treatment increases the de novo synthesis of CD1d. This change in surface CD1d expression was functionally relevant, as IFN-beta-treated dendritic cells are more efficient in stimulating invariant NKT cells than untreated controls. Taken together, these data support a role for early IFN-beta-mediated up-regulation of CD1d in NKT cell activation during infection. PMID- 17114456 TI - Heat shock protein 70 is secreted from tumor cells by a nonclassical pathway involving lysosomal endosomes. AB - Heat shock protein (HSP)70 can be released from tumor cells and stimulate a potent antitumor immune response. However, HSP70 does not contain a consensus secretory signal and thus cannot traverse the plasma membrane by conventional mechanisms. We have observed HSP70 release from intact human prostate carcinoma cell lines (PC-3 and LNCaP) by a mechanism independent of de novo HSP70 synthesis or cell death. This pathway is similar to one used by the leaderless protein IL 1beta. Our studies show that HSP70 release involves transit though an endolysosomal compartment and is inhibited by lysosomotropic compounds. In addition, the rate of HSP70 secretion correlates well with the appearance of the lysosomal marker LAMP1 on the cell surface, further suggesting the role for endolysosomes. The entry of HSP70 into this secretory compartment appears to involve the ABC family transporter proteins and ABC transporter inhibitor glibenclamide antagonizes secretion. Although the cell signals involved in triggering stress induced HSP70 release though this lysosomal pathway are largely unknown, our experiments suggest a regulatory role for extracellular ATP. These mechanisms appear to be shared by IL-1beta secretion. Following release, we observed the binding of extracellular HSP70 to the cell surface of the prostate carcinoma cells. These findings suggest that secreted HSP70 can take part in paracrine or autocrine interactions with adjacent cell surfaces. Our experiments therefore suggest a mechanism for HSP70 secretion and binding to the surface of other cells that may be involved in recognition of the tumor cells by the immune system. PMID- 17114457 TI - Spontaneous large-scale lymphoid neogenesis and balanced autoimmunity versus tolerance in the stomach of H+/K+-ATPase-reactive TCR transgenic mouse. AB - Autoimmunity is often accompanied by the development of ectopic lymphoid tissues in the target organ, and these tissues have been believed to have close relevance to the severity of the disease. However, the true relationship between the extent of such lymphoid structures and the intensity or type of immune responses mediated by self-reactive T cells has remained unclear. In the present study, we generated transgenic mice expressing TCR from an autoimmune gastritis (AIG) inducing Th1 cell clone specific for one of the major stomach self-Ags, H(+)/K(+) ATPase alpha subunit. The transgenic mice spontaneously develop massive lymphoid neogenesis with a highly organized tissue structure in the gastric mucosa, demonstrating Ag-specific, T cell-mediated induction of the lymphoid tissues. Nevertheless, the damage of surrounding tissue and autoantibody production were considerably limited compared with those in typical AIG induced by neonatal thymectomy. Such a moderate pathology is likely due to the locally restricted activation and Th2 skewing of self-reactive T cells, as well as the accumulation of naturally occurring regulatory T cells in the target organ. Altogether, the findings suggest that lymphoid neogenesis in chronic autoimmunity does not simply correlate with the destructive response; rather, the overall activation status of the T cell network, i.e., the balance of self-reactivity and tolerance, in the local environment has an impact. PMID- 17114458 TI - Microarrays reveal distinct gene signatures in the thymus of seropositive and seronegative myasthenia gravis patients and the role of CC chemokine ligand 21 in thymic hyperplasia. AB - Myasthenia gravis (MG) is an autoimmune disease mainly caused by antiacetylcholine receptor autoantibodies (seropositive (SP) disease) or by Abs against unknown autoantigenic target(s) (seronegative (SN) disease). Thymectomy is usually beneficial although thymic hyperplasia with ectopic germinal centers is mainly observed in SP MG. To understand the role of thymus in the disease process, we compared the thymic transcriptome of non-MG adults to those of SP patients with a low or high degree of hyperplasia or SN patients. Surprisingly, an overexpression of MHC class II, Ig, and B cell marker genes is observed in SP but also SN MG patients. Moreover, we demonstrate an overexpression of CXCL13 in all MG thymuses leading probably to the generalized B cell infiltration. However, we find different chemotactic properties for MG subgroups and, especially, a specific overexpression of CCL21 in hyperplastic thymuses triggering most likely ectopic germinal center development. Besides, SN patients present a peculiar signature with an abnormal expression of genes involved in muscle development and synaptic transmission, but also genes implicated in host response, suggesting that viral infection might be related to SN MG. Altogether, these results underline differential pathogenic mechanisms in the thymus of SP and SN MG and propose new research areas. PMID- 17114459 TI - General nature of the STAT3-activated anti-inflammatory response. AB - Although many cytokine receptors generate their signals via the STAT3 pathway, the IL-10R appears unique in promoting a potent anti-inflammatory response (AIR) via STAT3 to antagonize proinflammatory signals that activate the innate immune response. We found that heterologous cytokine receptor systems that activate STAT3 but are naturally refractory (the IL-22R), or engineered to be refractory (the IL-6, leptin, and erythropoietin receptors), to suppressor of cytokine signaling-3-mediated inhibition activate an AIR indistinguishable from IL-10. We conclude that the AIR is a generic cytokine signaling pathway dependent on STAT3 but not unique to the IL-10R. PMID- 17114460 TI - Nucleocytoplasmic shuttling of HMGB1 is regulated by phosphorylation that redirects it toward secretion. AB - The high mobility group box 1 (HMGB1) protein can be secreted by activated monocytes and macrophages and functions as a late mediator of sepsis. HMGB1 contains two nuclear localization signals (NLSs) for controlled nuclear transport, and acetylation of both NLSs of HMGB1 is involved in nuclear transport toward secretion. However, phosphorylation of HMGB1 and its relation to nuclear transport have not been shown. We show here that HMGB1 is phosphorylated and dynamically shuttled between cytoplasmic and nuclear compartments according to its phosphorylation state. Phosphorylation of HMGB1 was detected by metabolic labeling and Western blot analysis after treatments with TNF-alpha and okadaic acid, a phosphatase inhibitor. Hyperphosphorylated HMGB1 in RAW 264.7 and human monocytes was relocated to the cytoplasm. In a nuclear import assay, phosphorylated HMGB1 in the cytoplasm did not enter the nucleus. We mutated serine residues of either or both NLSs of HMGB1 to glutamic acid to simulate a phosphorylated state and examined the binding of HMGB1 to karyopherin-alpha1, which was identified as the nuclear import protein for HMGB1 in this study. Substitution to glutamic acid in either NLSs decreased the binding with karyopherin-alpha1 by approximately 50%; however, substitution of both NLSs showed no binding, and HMGB1 was relocated to the cytoplasm and subsequently secreted. These data support the hypothesis that HMGB1 could be phosphorylated and that the direction of transport is regulated by phosphorylation of both NLS regions. PMID- 17114461 TI - IFN regulatory factor 4 and 8 promote Ig light chain kappa locus activation in pre-B cell development. AB - Previous studies have shown that B cell development is blocked at the pre-B cell stage in IFN regulatory factor (IRF)4 (pip) and IRF8 (IFN consensus sequence binding protein) double mutant mice (IRF4,8(-/-)). In this study, the molecular mechanism by which IRF4,8 regulate pre-B cell development was further investigated. We show that IRF4,8 function in a B cell intrinsic manner to control pre-B cell development. IRF4,8(-/-) mice expressing a Bcl-2 transgene fail to rescue pre-B cell development, suggesting that the defect in B cell development in IRF4,8(-/-) mice is not due to a lack of survival signal. IRF4,8( /-) pre-B cells display a high proliferation index that may indirectly inhibit the L chain rearrangement. However, forced cell cycle exit induced by IL-7 withdrawal fails to rescue the development of IRF4,8(-/-) pre-B cells, suggesting that cell cycle exit by itself is not sufficient to rescue the development of IRF4,8(-/-) pre-B cells and that IRF4,8 may directly regulate the activation of L chain loci. Using retroviral mediated gene transduction, we show that IRF4 and IRF8 function redundantly to promote pre-B cell maturation and the generation of IgM(+) B cells. Molecular analysis indicates that IRF4, when expressed in IRF4,8( /-) pre-B cells, induces kappa germline transcription, enhances V(D)J rearrangement activity at the kappa locus, and promotes L chain rearrangement and transcription. Chromatin immunoprecipitation assay further reveals that IRF4 expression leads to histone modifications and enhanced chromatin accessibility at the kappa locus. Thus, IRF4,8 control pre-B cell development, at least in part, by promoting the activation of the kappa locus. PMID- 17114462 TI - Allergy-driven alternative splicing of IL-13 receptor alpha2 yields distinct membrane and soluble forms. AB - IL-13 is a key mediator of allergic inflammation. Its diverse functions are mediated by a complex receptor system including IL-4Ralpha, IL-13Ralpha1, and IL 13Ralpha2. IL-4Ralpha and IL-13Ralpha1 form a high-affinity signaling heterodimer. IL-13Ralpha2 binds IL-13 with high affinity and has been found to exist in membrane and soluble forms. Soluble IL-13Ralpha2 has been postulated as a critical endogenous modulator of IL-13 responses. However, the mechanism of generation for the soluble form remains unclear. We present the initial study that a mechanism for generation of the soluble form is alternative splicing and that alternative splicing yields a distinct form of soluble IL-13Ralpha2. We found that several mouse organs expressed two IL-13Ralpha2 transcripts, the 1152 bp transcript encoding the full-length protein and the 1020-bp transcript lacking exon10, which encodes the transmembrane region. Deletion of exon 10 (DeltaEx10) caused a frameshift resulting in a different amino acid sequence from position 327 to position 339 and early termination. Constructs encoding both splice variants were transfected into WEHI-274.1 cells. Transfectants expressing the full-length transcript had IL-13Ralpha2 on the cell surface but produced minimal soluble IL-13Ralpha2 in the supernatants. In contrast, transfectants expressing the DeltaEx10 transcript displayed no membrane IL-13Ralpha2 but secreted high levels of soluble IL-13Ralpha2 capable of inhibiting IL-13 signaling. Both variants bound IL-13, but the DeltaEx10 variant displayed approximately 2-fold increase in IL-13 binding activity. Expression of the two IL-13Ralpha2 transcripts was differentially regulated in vivo in an experimental allergic asthma model. Thus, alternatively spliced variants of IL-13Ralpha2 may have a distinct biologic function in vivo. PMID- 17114463 TI - Analysis of the individual contributions of Igalpha (CD79a)- and Igbeta (CD79b) mediated tonic signaling for bone marrow B cell development and peripheral B cell maturation. AB - The individual contribution of Igalpha and Igbeta for BCR-triggered fates is unclear. Prior evidence supports conflicting ideas concerning unique as well as redundant functions for these proteins in the context of BCR/pre-BCR signaling. Part of this ambiguity may reflect the recent appreciation that Igalpha and Igbeta participate in both Ag-independent (tonic) and Ag-dependent signaling. The present study undertook defining the individual requirement for Igalpha and Igbeta under conditions where only ligand-independent tonic signaling was operative. In this regard, we have constructed chimeric proteins containing one or two copies of the cytoplasmic domains of either Igalpha or Igbeta and Igalpha/Igbeta heterodimers with targeted Tyr-->Phe modifications. The ability of these proteins to act as surrogate receptors and trigger early bone marrow and peripheral B cell maturation was tested in RAG2(-/-) primary pro-B cell lines and in gene transfer experiments in the muMT mouse model. We considered that the threshold for a functional activity mediated by the pre-BCR/BCR might only be reached when two functional copies of the Igalpha/Igbeta ITAM domain are expressed together, and therefore the specificity conferred by these proteins can only be observed in these conditions. We found that the ligand-independent tonic signal is sufficient to drive development into mature follicular B cells and both Igalpha and Igbeta chains supported formation of this population. In contrast, neither marginal zone nor B1 mature B cell subsets develop from bone marrow precursors under conditions where only tonic signals are generated. PMID- 17114464 TI - Mapping of quantitative trait loci determining NK cell-mediated resistance to MHC class I-deficient bone marrow grafts in perforin-deficient mice. AB - NK cells reject allogeneic and MHC class I-deficient bone marrow (BM) grafts in vivo. The mechanisms used by NK cells to mediate this rejection are not yet thoroughly characterized. Although perforin plays a major role, perforin independent mechanisms are involved as well. C57BL/6 mice deficient in perforin (B6 perforin knockout (PKO)) reject class I-deficient TAP-1 KO BM cells as efficiently as normal B6 mice. In contrast, perforin-deficient 129S6/SvEvTac mice (129 PKO) cannot mediate this rejection while normal 129 mice efficiently reject. This suggests that in 129, but not in B6, mice, perforin is crucial for NK cell mediated rejection of MHC class I-deficient BM grafts. To identify loci linked to BM rejection in perforin-deficient mice, we generated backcross 1 progeny by crossing (129 x B6)F(1) PKO mice to 129 PKO mice. In transplantation experiments, >350 backcross 1 progeny were analyzed and displayed a great variation in ability to reject TAP-1 KO BM grafts. PCR-based microsatellite mapping identified four quantitative trait loci (QTL) on chromosomes 2, 4, and 8, with the QTL on chromosome 8 showing the highest significance, as well as a fifth epistatic QTL on chromosome 3. This study describes the first important step toward identifying BM graft resistance gene(s). PMID- 17114465 TI - Gene expression profiling reveals unique pathways associated with differential severity of lyme arthritis. AB - The murine model of Lyme disease provides a unique opportunity to study the localized host response to similar stimulus, Borrelia burgdorferi, in the joints of mice destined to develop severe arthritis (C3H) or mild disease (C57BL/6). Pathways associated with the response to infection and the development of Lyme arthritis were identified by global gene expression patterns using oligonucleotide microarrays. A robust induction of IFN-responsive genes was observed in severely arthritic C3H mice at 1 wk of infection, which was absent from mildly arthritic C57BL/6 mice. In contrast, infected C57BL/6 mice displayed a novel expression profile characterized by genes involved in epidermal differentiation and wound repair, which were decreased in the joints of C3H mice. These expression patterns were associated with disease state rather than inherent differences between C3H and C57BL/6 mice, because C57BL/6-IL-10(-/-) mice infected with B. burgdorferi develop more severe arthritis than C57BL/6 mice and displayed an early gene expression profile similar to C3H mice. Gene expression profiles at 2 and 4 wk postinfection revealed a common response of all strains that was likely to be important for the host defense to B. burgdorferi and mediated by NF-kappaB-dependent signaling. The gene expression profiles identified in this study add to the current understanding of the host response to B. burgdorferi and identify two novel pathways that may be involved in regulating the severity of Lyme arthritis. PMID- 17114466 TI - Increased natural CD4+CD25+ regulatory T cells and their suppressor activity do not contribute to mortality in murine polymicrobial sepsis. AB - Regulatory T cells (Tregs), including natural CD4+CD25+ Tregs and inducible IL-10 producing T regulatory type 1 (T(R)1) cells, maintain tolerance and inhibit autoimmunity. Recently, increased percentages of Tregs have been observed in the blood of septic patients, and ex vivo-activated Tregs were shown to prevent polymicrobial sepsis mortality. Whether endogenous Tregs contribute to sepsis outcome remains unclear. Polymicrobial sepsis, induced by cecal ligation and puncture, caused an increased number of splenic Tregs compared with sham-treated mice. Splenic CD4+CD25+ T cells from septic mice expressed higher levels of Foxp3 mRNA and were more efficient suppressors of CD4+CD25- T effector cell proliferation. Isolated CD4+ T cells from septic mice displayed increased intracellular IL-10 staining following stimulation, indicating that T(R)1 cells may also be elevated in sepsis. Surprisingly, Ab depletion of total CD4+ or CD4+CD25+ populations did not affect mortality. Furthermore, no difference in survival outcome was found between CD25 or IL-10 null mice and wild-type littermates, indicating that Treg or T(R)1-generated IL-10 are not required for survival. These results demonstrate that, although sepsis causes a relative increase in Treg number and increases their suppressive function, their presence does not contribute significantly to overall survival in this model. PMID- 17114467 TI - TLR4-dependent NF-kappaB activation and mitogen- and stress-activated protein kinase 1-triggered phosphorylation events are central to Helicobacter pylori peptidyl prolyl cis-, trans-isomerase (HP0175)-mediated induction of IL-6 release from macrophages. AB - Helicobacter pylori infection is associated with the local production of chemokines and cytokines, of which IL-6 is overexpressed at the margin of gastric ulcer in H. pylori-positive gastritis. Cells of the monocytic lineage are the major sources of IL-6, and mononuclear cell infiltration in the lamina propria is characteristic of H. pylori-induced chronic infection. Our study shows for the first time that a secreted peptidyl prolyl cis-, trans-isomerase, HP0175 elicits IL-6 gene expression and IL-6 release from macrophages. An isogenic strain inactivated in the HP0175 gene (knockout) was attenuated in its IL-6-inducing ability, which was restored after complementation with the HP0175 gene. The specificity of the HP0175-induced effect was confirmed by the fact that rHP0175 purified from HEK293 cells could also induce IL-6 release, ruling out the possibility that the observed effect was due to bacterial contaminants. HP0175 was capable of interacting directly with the extracellular domain of TLR4. HP0175 induced IL-6 gene expression was critically dependent on TLR4-dependent NF-kappaB and MAPK activation. TLR4/PI3K-dependent ERK1/2 and p38 MAPK signaling converged upon activation of mitogen- and stress-activated protein kinase 1 (MSK1). The central role of MSK1 was borne out by the fact that silencing of MSK1 expression abrogated HP0175-mediated NF-kappaB-dependent IL-6 gene transcription. MSK1 regulated the recruitment of p65 and phopho-Ser(10)-histone H3 to the IL-6 promoter. HP0175 therefore regulated IL-6 gene transcription through chromatin modification at the IL-6 promoter. PMID- 17114468 TI - Human Langerhans cells express a specific TLR profile and differentially respond to viruses and Gram-positive bacteria. AB - Dendritic cells (DC) are APCs essential for the development of primary immune responses. In pluristratified epithelia, Langerhans cells (LC) are a critical subset of DC which take up Ags and migrate toward lymph nodes upon inflammatory stimuli. TLR allow detection of pathogen-associated molecular patterns (PAMP) by different DC subsets. The repertoire of TLR expressed by human LC is uncharacterized and their ability to directly respond to PAMP has not been systematically investigated. In this study, we show for the first time that freshly purified LC from human skin express mRNA encoding TLR1, TLR2, TLR3, TLR5, TLR6 and TLR10. In addition, keratinocytes ex vivo display TLR1-5, TLR7, and TLR10. Accordingly, highly enriched immature LC efficiently respond to TLR2 agonists peptidoglycan and lipoteichoic acid from Gram-positive bacteria, and to dsRNA which engages TLR3. In contrast, LC do not directly sense TLR7/8 ligands and LPS from Gram-negative bacteria, which signals through TLR4. TLR engagement also results in cytokine production, with marked differences depending on the PAMP detected. TLR2 and TLR3 ligands increase IL-6 and IL-8 production, while dsRNA alone stimulates TNF-alpha release. Strikingly, only peptidoglycan triggers IL-10 secretion, thereby suggesting a specific function in tolerance to commensal Gram-positive bacteria. However, LC do not produce IL-12p70 or type I IFNs. In conclusion, human LC are equipped with TLR that enable direct detection of PAMP from viruses and Gram-positive bacteria, subsequent phenotypic maturation, and differential cytokine production. This implies a significant role for LC in the control of skin immune responses. PMID- 17114469 TI - The role of TCR engagement and activation-induced cell death in sepsis-induced T cell apoptosis. AB - Sepsis induces extensive apoptosis in T and B cells suggesting that the loss of immune effector cells could be one explanation for the profound immunosuppression observed in this disorder. Unfortunately, the mechanisms responsible for lymphocyte apoptosis in sepsis remain unknown. In T cells, apoptosis can occur through activation-induced cell death (AICD) in which engagement of the Ag receptors by cognate Ag or polyclonal activators such as bacteria-derived superantigens induces activation, proliferation, and apoptosis. We examined whether proliferation and AICD are necessary for apoptotic cell death in sepsis using normal and TCR transgenic mice. Results show that although sepsis resulted in activation of a small percentage of T cells, no proliferation was detected during the first 48 h following onset, a time when extensive apoptosis is observed. We also observed that T cells do not enter the cell cycle, and stimulation via the TCR in TCR transgenic animals does not enhance or decrease cell death in sepsis. Interestingly, T cells recovered from septic mice retained their ability to proliferate and synthesize cytokines albeit at reduced levels. With the exception of IL-10, which was increased in lymphocytes from mice with sepsis, sepsis caused a decrease in the production of both proinflammatory and anti-inflammatory cytokines. We conclude that lymphocyte apoptosis in sepsis does not require proliferation, TCR engagement, or AICD. Thus the immunosuppression observed in sepsis cannot be the result of T cell deletion via the TCR. PMID- 17114470 TI - Antigen-specific CD8+ T cells respond to Chlamydia trachomatis in the genital mucosa. AB - Following sexual transmission, Chlamydia trachomatis specifically targets genital tract epithelial cells. Because epithelial cells are readily recognized by CD8+ T cells, the response of CD8+ T cells to Chlamydia infection has been explored in a number of studies. It has been shown that CD8+ T cells are present in the genital tracts of mice following C. trachomatis infection, but the specificity of these T cells has remained undefined. To determine whether Chlamydia-specific CD8+ T cells migrate to the genital tract in response to Chlamydia infection, we generated retrogenic mice that express a TCR specific for a Chlamydia-specific T cell Ag CrpA. T cells from the retrogenic mice were transferred into naive recipient animals to increase the frequency of Chlamydia-specific T cells to a level at which they could be tracked during primary infection. We observed that the Chlamydia-specific retrogenic T cells proliferated in lymph nodes draining the genital tract in response to genital infection with C. trachomatis. Furthermore, we found that these cells acquired the ability to produce IFN-gamma and migrated into the genital mucosa of the infected mice. PMID- 17114471 TI - TLR4 mediates vaccine-induced protective cellular immunity to Bordetella pertussis: role of IL-17-producing T cells. AB - Whole cell pertussis vaccines (Pw) induce Th1 responses and protect against Bordetella pertussis infection, whereas pertussis acellular vaccines (Pa) induce Ab and Th2-biased responses and also protect against severe disease. In this study, we show that Pw failed to generate protective immunity in TLR4-defective C3H/HeJ mice. In contrast, protection induced with Pa was compromised, but not completely abrogated, in C3H/HeJ mice. Immunization with Pw, but not Pa, induced a population of IL-17-producing T cells (Th-17), as well as Th1 cells. Ag specific IL-17 and IFN-gamma production was significantly lower in Pw-immunized TLR4-defective mice. Furthermore, treatment with neutralizing anti-IL-17 Ab immediately before and after B. pertussis challenge significantly reduced the protective efficacy of Pw. Stimulation of dendritic cells (DC) with Pw promoted IL-23, IL-12, IL-1beta, and TNF-alpha production, which was impaired in DC from TLR4-defective mice. B. pertussis LPS, which is present in high concentrations in Pw, induced IL-23 production by DC, which enhanced IL-17 secretion by T cells, but the induction of Th-17 cells was also dependent on IL-1. In addition, we identified a new effector function for IL-17, activating macrophage killing of B. pertussis, and this bactericidal activity was less efficient in macrophages from TLR4-defective mice. These data provide the first definitive evidence of a role for TLRs in protective immunity induced by a human vaccine. Our findings also demonstrate that activation of innate immune cells through TLR4 helps to direct the induction of Th1 and Th-17 cells, which mediate protective cellular immunity to B. pertussis. PMID- 17114472 TI - Interleukin-8 induction by Helicobacter pylori in gastric epithelial cells is dependent on apurinic/apyrimidinic endonuclease-1/redox factor-1. AB - Helicobacter pylori infection causes inflammation and increases the expression of IL-8 in human gastric epithelial cells. H. pylori activates NF-kappaB and AP-1, essential transcriptional factors in H. pylori-induced IL-8 gene transcription. Although colonization creates a local oxidative stress, the molecular basis for the transition from infection to the expression of redox-sensitive cytokine genes is unknown. We recently reported that the expression of apurinic/apyrimidinic endonuclease-1/redox factor-1 (APE-1/Ref-1), which repairs oxidative DNA damage and reductively activates transcription factors including AP-1 and NF-kappaB, is increased in human gastric epithelia during H. pylori infection. In this study, we examine whether APE-1/Ref-1 functions in the modulation of IL-8 gene expression in H. pylori-infected human gastric epithelial cells. Small interfering RNA-mediated silencing of APE-1/Ref-1 inhibited basal and H. pylori induced AP-1 and NF-kappaB DNA-binding activity without affecting the nuclear translocation of these transcription factors and also reduced H. pylori-induced IL-8 mRNA and protein. In contrast, overexpression of APE-1/Ref-1 enhanced basal and H. pylori-induced IL-8 gene transcription, and the relative involvement of AP 1 in inducible IL-8 promoter activity was greater in APE-1/Ref-1 overexpressing cells than in cells with basal levels of APE-1/Ref-1. APE-1/Ref-1 inhibition also reduced other H. pylori-induced chemokine expression. By implicating APE-1/Ref-1 as an important regulator of gastric epithelial responses to H. pylori infection, these data elucidate a novel mechanism controlling transcription and gene expression in bacterial pathogenesis. PMID- 17114473 TI - Pseudomonas aeruginosa-induced human mast cell apoptosis is associated with up regulation of endogenous Bcl-xS and down-regulation of Bcl-xL. AB - Mast cells play a critical role in the host defense against bacterial infection. Recently, apoptosis has been demonstrated to be essential in the regulation of host response to Pseudomonas aeruginosa. In this study we show that human mast cell line HMC-1 and human cord blood-derived mast cells undergo apoptosis as determined by the ssDNA formation after infection with P. aeruginosa. P. aeruginosa induced activation of caspase-3 in mast cells as evidenced by the cleavage of D4-GDI, an endogenous caspase-3 substrate and the generation of an active form of caspase-3. Interestingly, P. aeruginosa treatment induced up regulation of Bcl-x(S) and down-regulation of Bcl-x(L). Bcl-x(S), and Bcl-x(L) are alternative variants produced from the same Bcl-x pre-mRNA. The former is proapoptotic and the latter is antiapoptotic likely through regulating mitochondrial membrane integrity. Treatment of mast cells with P. aeruginosa induced release of cytochrome c from mitochondria and loss of mitochondrial membrane potentials. Moreover, P. aeruginosa treatment reduced levels of Fas associated death domain protein-like IL-1beta-converting enzyme-inhibitory proteins (FLIPs) that are endogenous apoptosis inhibitors through counteraction with caspase-8. Thus, human mast cells undergo apoptosis after encountering P. aeruginosa through a mechanism that likely involves both the Bcl family protein mitochondrial-dependent and the FLIP-associated caspase-8 pathways. PMID- 17114474 TI - The IFN-independent response to virus particle entry provides a first line of antiviral defense that is independent of TLRs and retinoic acid-inducible gene I. AB - The innate immune system responds to pathogen infection by eliciting a nonspecific immune response following the recognition of various pathogen associated molecular patterns. TLRs and the RNA helicases retinoic acid-inducible gene I (RIG-I) and melanoma differentiation-associated gene 5 recognize foreign nucleic acid within endosomal and cytoplasmic compartments, respectively, initiating a signaling cascade that involves the induction of type I IFN through the transcription factors IFN regulatory factor (IRF) 3 and NF-kappaB. However, a recent paradigm has emerged in which bacterial DNA and double-stranded B-form DNA trigger type I IFN production through an uncharacterized TLR- and RIG-I independent pathway. We have previously described a response in primary fibroblasts wherein the entry of diverse RNA- and DNA-enveloped virus particles is sufficient to induce a subset of IFN-stimulated genes and a complete antiviral response in an IRF3-dependent, IFN-independent manner. In this study, we show that the innate immune response to virus particle entry is independent of both TLR and RIG-I pathways, confirming the existence of novel innate immune mechanisms that result in the activation of IRF3. Furthermore, we propose a model of innate antiviral immunity in which exposure to increasing numbers of virus particles elevates the complexity of the cellular response from an intracellular, IFN-independent response to one involving secretion of cytokines and activation of infiltrating immune cells. PMID- 17114475 TI - A new staphylococcal anti-inflammatory protein that antagonizes the formyl peptide receptor-like 1. AB - Bacteria have developed mechanisms to escape the first line of host defense, which is constituted by the recruitment of phagocytes to the sites of bacterial invasion. We previously described the chemotaxis inhibitory protein of Staphylococcus aureus, a protein that blocks the activation of neutrophils via the formyl peptide receptor (FPR) and C5aR. We now describe a new protein from S. aureus that impaired the neutrophil responses to FPR-like1 (FPRL1) agonists. FPRL1 inhibitory protein (FLIPr) inhibited the calcium mobilization in neutrophils stimulated with MMK-1, WKYMVM, prion-protein fragment PrP(106-126), and amyloid beta(1-42). Stimulation with low concentrations of fMLP was partly inhibited. Directed migration was also completely prevented toward MMK-1 and partly toward fMLP. Fluorescence-labeled FLIPr efficiently bound to neutrophils, monocytes, B cells, and NK cells. HEK293 cells transfected with human C5aR, FPR, FPRL1, and FPRL2 clearly showed that FLIPr directly bound to FPRL1 and, at higher concentrations, also to FPR but not to C5aR and FPRL2. FLIPr can reveal unknown inflammatory ligands crucial during S. aureus infections. As a novel described FPRL1 antagonist, it might lead to the development of therapeutic agents in FPRL1 mediated inflammatory components of diseases such as systemic amyloidosis, Alzheimer's, and prion disease. PMID- 17114476 TI - Direct CD28 costimulation is required for CD8+ T cell-mediated resistance to an acute viral disease in a natural host. AB - Previous studies have suggested that, differing from model Ags, viruses that replicate extensively in the host still induce normal CD8+ T cell responses in the absence of CD28 costimulation. Because these studies were performed with viruses that do not normally cause acute disease, an important remaining question is whether CD28 costimulation is required for CD8+ T cell-mediated resistance to widely replicating but pathogenic viruses. To address this question, we studied the role of CD28 costimulation in CD8+ T cell-mediated resistance to mousepox, a disease of the mouse caused by the natural mouse pathogen, the ectromelia virus (ECTV). C57BL/6 (B6) mice are naturally resistant to mousepox, partly due to a fast and strong CD8+ T cell response. We found that B6 mice deficient in CD28 (CD28 knockout (KO)) are highly susceptible to lethal mousepox during the early stages of ECTV infection but can be protected by immunization with the antigenically related vaccinia virus (VACV) or by adoptive transfer of CD28 KO anti-VACV memory CD8+ cells. Of interest, a thorough comparison of the CD8+ T cell responses to ECTV and VACV suggests that the main reason for the susceptibility of CD28 KO mice to mousepox is a reduced response at the early stages of infection. Thus, while in the absence of CD28 costimulation the end point strength of the T cell responses to nonpathogenic viruses may appear normal, CD28 costimulation increases the speed of the T cell response and is essential for resistance to a life-threatening acute viral disease. PMID- 17114477 TI - An HLA-A2.1-transgenic rabbit model to study immunity to papillomavirus infection. AB - We have established several HLA-A2.1-transgenic rabbit lines to provide a host to study CD8(+) T cell responses during virus infections. HLA-A2.1 protein expression was detected on cell surfaces within various organ tissues. Continuous cultured cells from these transgenic rabbits were capable of presenting both endogenous and exogenous HLA-A2.1-restricted epitopes to an HLA-A2.1-restricted epitope-specific CTL clone. A DNA vaccine containing an HLA-A2.1-restricted human papillomavirus type 16 E7 epitope (amino acid residues 82-90) stimulated epitope specific CTLs in both PBLs and spleen cells of transgenic rabbits. In addition, vaccinated transgenic rabbits were protected against infection with a mutant cottontail rabbit papillomavirus DNA containing an embedded human papillomavirus type 16 E7/82-90 epitope. Complete protection was achieved using a multivalent epitope DNA vaccine based on epitope selection from cottontail rabbit papillomavirus E1 using MHC class I epitope prediction software. HLA-A2.1 transgenic rabbits will be an important preclinical animal model system to study virus-host interactions and to assess specific targets for immunotherapy. PMID- 17114478 TI - The tumor-promoting effect of TNF-alpha involves the induction of secretory leukocyte protease inhibitor. AB - According to the cancer immunoediting concept, inflammatory mediators play not only a critical role in promoting host protection against cancer but also contribute to cancer cell growth and survival. TNF-alpha is a critical factor in this network. However, the mechanisms underlying the tumor-promoting effect of TNF-alpha have not been fully elucidated yet. We previously reported that in vitro culture of Lewis lung carcinoma 3LL cells with TNF-alpha-producing macrophages resulted in enhanced resistance toward TNF-alpha-mediated lysis and increased malignancy of the 3LL cells. In this study, we analyzed the effects of endogenous TNF-alpha on TNF-alpha resistance and malignant behavior in vivo of low-malignant/TNF-alpha-sensitive 3LL-S cells and cancer cells derived from 3LL-S tumors that developed in wild-type or TNF-alpha(-/-) mice. Interestingly, 3LL-S cells acquired a malignant phenotype in vivo depending on the presence of host TNF-alpha, whereas acquisition of TNF-alpha resistance was TNF-alpha-independent. This result suggested that malignancy-promoting characteristics of 3LL-S cells other than TNF-alpha resistance are influenced in vivo by TNF-alpha. We previously identified the malignancy-promoting genes, secretory leukocyte protease inhibitor (SLPI) and S100A4, as being up-regulated in 3LL-S cells upon their s.c. growth in wild-type mice. In this study, we show that SLPI, but not S100A4, was induced in 3LL-S cells both in vitro and in vivo by TNF-alpha, and that silencing of in vivo induced 3LL-S SLPI expression using RNA interference abrogated in vivo progression but did not influence TNF-alpha resistance. These data indicate that SLPI induction may be one mechanism whereby TNF-alpha acts as an endogenous tumor promoter. PMID- 17114479 TI - CXCL12 limits inflammation by localizing mononuclear infiltrates to the perivascular space during experimental autoimmune encephalomyelitis. AB - The inflammatory response in the CNS begins with the movement of leukocytes across the blood-brain barrier in a multistep process that requires cells to pass through a perivascular space before entering the parenchyma. The molecular mechanisms that orchestrate this movement are not known. The chemokine CXCL12 is highly expressed throughout the CNS by microendothelial cells under normal conditions, suggesting it might play a role maintaining the blood-brain barrier. We tested this hypothesis in the setting of experimental autoimmune encephalomyelitis (EAE) by using AMD3100, a specific antagonist of the CXCL12 receptor CXCR4. We demonstrate that the loss of CXCR4 activation enhances the migration of infiltrating leukocytes into the CNS parenchyma. CXCL12 is expressed at the basolateral surface of CNS endothelial cells in normal spinal cord and at the onset of EAE. This polarity is lost in vessels associated with an extensive parenchymal invasion of mononuclear cells during the peak of disease. Inhibition of CXCR4 activation during the induction of EAE leads to loss of the typical intense perivascular cuffs, which are replaced with widespread white matter infiltration of mononuclear cells, worsening the clinical severity of the disease and increasing inflammation. Taken together, these data suggest a novel anti inflammatory role for CXCL12 during EAE in that it functions to localize CXCR4 expressing mononuclear cells to the perivascular space, thereby limiting the parenchymal infiltration of autoreactive effector cells. PMID- 17114480 TI - Attenuating burn wound inflammatory signaling reduces systemic inflammation and acute lung injury. AB - The relationship between local inflammation and the subsequent systemic inflammatory response is poorly described. In a burn injury model, the dermal inflammatory response may act as an ongoing trigger for the systemic inflammatory response syndrome (SIRS) and subsequent systemic complications. We hypothesized that topical attenuation of burn wound inflammatory signaling will control the dermal inflammatory source, attenuate SIRS, and reduce acute lung injury. Mice received a 30% total body surface area burn. Subgroups were treated with specific p38 MAPK inhibitor or vehicle, which was topically applied to wounds. Topical p38 MAPK inhibition significantly reduced burn wound inflammatory signaling and subsequent systemic expression of proinflammatory cytokines and chemokines. In vitro macrophage functional assays demonstrated a significant attenuation in serum inflammatory mediators from animals receiving the topical inhibitor. Topical p38 MAPK inhibition resulted in significantly less pulmonary inflammatory response via reduction of pulmonary neutrophil sequestration, pulmonary cytokine expression, and a significant reduction in pulmonary microvascular injury and edema formation. Although dermal activating transcription factor-2, a downstream p38 MAPK target, was significantly reduced, there was no reduction in pulmonary activating transcription factor-2 expression, arguing against significant systemic absorption of the topical inhibitor. These experiments demonstrate a strong interaction between dermal inflammation and systemic inflammatory response. Attenuating local inflammatory signaling appears effective in reducing SIRS and subsequent systemic complications after burn injury. PMID- 17114481 TI - Macrophage migration inhibitory factor induces macrophage recruitment via CC chemokine ligand 2. AB - Macrophage migration inhibitory factor (MIF) was originally identified for its ability to inhibit the random migration of macrophages in vitro. MIF is now recognized as an important mediator in a range of inflammatory disorders. We recently observed that the absence of MIF is associated with a reduction in leukocyte-endothelial cell interactions induced by a range of inflammatory mediators, suggesting that one mechanism whereby MIF acts during inflammatory responses is by promoting leukocyte recruitment. However, it is unknown whether MIF is capable of inducing leukocyte recruitment independently of additional inflammatory stimuli. In this study, we report that MIF is capable of inducing leukocyte adhesion and transmigration in postcapillary venules in vivo. Moreover, leukocytes recruited in response to MIF were predominantly CD68(+) cells of the monocyte/macrophage lineage. Abs against the monocyte-selective chemokine CCL2 (JE/MCP-1) and its receptor CCR2, but not CCL3 and CXCL2, significantly inhibited MIF-induced monocyte adhesion and transmigration. CCL2(-/-) mice displayed a similar reduction in MIF-induced recruitment indicating a critical role of CCL2 in the MIF-induced response. This hypothesis was supported by findings that MIF induced CCL2 release from primary microvascular endothelial cells. These data demonstrate a previously unrecognized function of this pleiotropic cytokine: induction of monocyte migration into tissues. This function may be critical to the ability of MIF to promote diseases such as atherosclerosis and rheumatoid arthritis, in which macrophages are key participants. PMID- 17114482 TI - The differing roles of the classical and mannose-binding lectin complement pathways in the events following skeletal muscle ischemia-reperfusion. AB - Complement is an important mediator of the injuries observed after skeletal muscle ischemia and subsequent reperfusion. Although the classical pathway had been assumed to be the major pathway of activation leading to injury, the mannose binding lectin (MBL) pathway might also play a contributing role. In this study, we found that MBL-deficient mice had significant protection after skeletal muscle reperfusion injury compared with wild-type, classical pathway-specific C1q deficient mice, or MBL-deficient mice reconstituted with recombinant human MBL. MBL-deficient mice, however, were not protected from permeability edema or secondary lung injury after ischemia-reperfusion. These data indicate that blockade of the classical pathway alone (C1q) is protective against permeability edema and remote pulmonary injury but not protective against histologic muscle injury. In contrast, blocking the MBL pathway alone protects against histological injury but is not protective against permeability edema or lung injury. Thus, the activation of both pathways is likely responsible for the full spectrum of injuries observed after skeletal muscle reperfusion injury. PMID- 17114483 TI - The Duffy antigen modifies systemic and local tissue chemokine responses following lipopolysaccharide stimulation. AB - The Duffy blood group Ag (dfy) binds selective CXC and CC chemokines at high affinity and is expressed on erythrocytes and endothelial cells. However, it does not transmit a signal via G proteins, as occurs with other seven-transmembrane receptors. We hypothesized that dfy functions as a chemokine reservoir and regulates inflammation by altering soluble chemokine concentrations in the blood and tissue compartments. We determined whether Duffy Ag "loss-of-function" phenotypes (human and murine) are associated with alterations in plasma chemokine concentrations during the innate inflammatory response to LPS. Plasma CXCL8 and CCL2 concentrations from humans homozygous for the GATA-1 box polymorphism, a dfy polymorphism that abrogates erythrocyte chemokine binding, were higher than in heterozygotes following LPS stimulation of their whole blood in vitro. Similarly, dfy(-/-) mice showed higher plasma MIP-2 concentrations than dfy(+/+) mice following LPS stimulation of whole blood in vitro. We then determined the relative contributions of erythrocyte and endothelial Duffy Ag in modifying chemokine concentrations and neutrophil recruitment in the lungs following intratracheal LPS administration in dfy(-/-) and dfy(+/+) mice reconstituted with dfy(-/-) or dfy(+/+) marrow. Mice lacking endothelial dfy expression had higher MIP-2 and keratinocyte chemoattractant concentrations in the airspaces. Mice lacking erythrocyte dfy had higher MIP-2 and keratinocyte chemoattractant concentrations in the lung tissue vascular space, but lower plasma chemokine concentrations associated with attenuated neutrophil recruitment into the airspaces. These data indicate that dfy alters soluble chemokine concentrations in blood and local tissue compartments and enhances systemic bioavailability of chemokines produced during local tissue inflammation. PMID- 17114484 TI - Translational regulation of autoimmune inflammation and lymphoma genesis by programmed cell death 4. AB - Both inflammatory diseases and cancer are associated with heightened protein translation. However, the mechanisms of translational regulation and the roles of translation factors in these diseases are not clear. Programmed cell death 4 (PDCD4) is a newly described inhibitor of protein translation. To determine the roles of PDCD4 in vivo, we generated PDCD4-deficient mice by gene targeting. We report here that mice deficient in PDCD4 develop spontaneous lymphomas and have a significantly reduced life span. Most tumors are of the B lymphoid origin with frequent metastasis to liver and kidney. However, PDCD4-deficient mice are resistant to inflammatory diseases such as autoimmune encephalomyelitis and diabetes. Mechanistic studies reveal that upon activation, PDCD4-deficient lymphocytes preferentially produce cytokines that promote oncogenesis but inhibit inflammation. These results establish that PDCD4 controls lymphoma genesis and autoimmune inflammation by selectively inhibiting protein translation in the immune system. PMID- 17114485 TI - A requirement for microglial TLR4 in leukocyte recruitment into brain in response to lipopolysaccharide. AB - To study the mechanisms involved in leukocyte recruitment induced by local bacterial infection within the CNS, we used intravital microscopy to visualize the interaction between leukocytes and the microvasculature in the brain. First, we showed that intracerebroventricular injection of LPS could cause significant rolling and adhesion of leukocytes in the brain postcapillary venules of wild type mice, while negligible recruitment was observed in TLR4-deficient C57BL/10ScCr mice and CD14 knockout mice, suggesting recruitment is mediated by TLR4/CD14-bearing cells. Moreover, we observed reduced but not complete inhibition of recruitment in MyD88 knockout mice, indicating both MyD88-dependent and -independent pathways are involved. The leukocyte recruitment responses in chimeric mice with TLR4-positive microglia and endothelium, but TLR4-negative leukocytes, were comparable to normal wild-type mice, suggesting either endothelium or microglia play a crucial role in the induction of leukocyte recruitment. LPS injection induced both microglial and endothelial activation in the CNS. Furthermore, minocycline, an effective inhibitor of microglial activation, completely blocked the rolling and adhesion of leukocytes in the brain and blocked TNF-alpha production in response to LPS in vivo. Minocycline did not affect activation of endothelium by LPS in vitro. TNFR p55/p75 double knockout mice also exhibited significant reductions in both rolling and adhesion in response to LPS, indicating TNF-alpha signaling is critical for the leukocyte recruitment. Our results identify a TLR4 detection system within the blood-brain barrier. The microglia play the role of sentinel cells detecting LPS thereby inducing endothelial activation and leading to efficient leukocyte recruitment to the CNS. PMID- 17114486 TI - Cyclooxygenase-2 gene transcription in a macrophage model of inflammation. AB - Infections involving LPS-bearing, Gram-negative bacteria can lead to acute inflammation and septic shock. Cyclooxygenase-2 (COX-2), the target of nonsteroidal anti-inflammatory drugs and selective COX-2 inhibitors, is importantly involved in these responses. We examined the dynamics of COX-2 gene expression in RAW264.7 murine macrophages treated with LPS as a model for COX-2 gene expression during inflammation. We established, using Northern blotting, nuclear run-on assays, and RT-PCR, that COX-2 transcriptional activation continues for at least 12 h after LPS treatment and involves at least three phases. Previous studies with murine macrophages identified an NF-kappaB site, a C/EBP site, and a cAMP response element-1 (CRE-1) as cis-acting elements in the COX-2 promoter. We identified three additional functional elements including a second CRE (CRE-2), an AP-1 site, and an E-box that overlaps CRE-1. The E-box mediates transcriptional repression whereas the other cis-elements are activating. Using electrophoretic mobility supershift and chromatin immunoprecipitation assays, we cataloged binding to each functional cis element and found them occupied to varying extents and by different transcription factors during the 12 h following LPS treatment. This suggests that the cis elements and their cognate transcription factors participate in a sequential, coordinated regulation of COX-2 gene expression during an inflammatory response. In support of this concept, we found, using inhibitors of Jun kinase and NF-kappaB p50 nuclear localization, that COX-2 gene transcription was completely dependent on phospho-c-Jun plus p50 at 6 h after LPS treatment but was only partially dependent on the combination of these factors at later treatment times. PMID- 17114487 TI - Neutrophils are a key component of the antitumor efficacy of topical chemotherapy with ingenol-3-angelate. AB - Harnessing neutrophils for the eradication of cancer cells remains an attractive but still controversial notion. In this study, we provide evidence that neutrophils are required to prevent relapse of skin tumors following topical treatment with a new anticancer agent, ingenol-3-angelate (PEP005). Topical PEP005 treatment induces primary necrosis of tumor cells, potently activates protein kinase C, and was associated with an acute T cell-independent inflammatory response characterized by a pronounced neutrophil infiltrate. In Foxn1(nu) mice depleted of neutrophils and in CD18-deficient mice (in which neutrophil extravasation is severely impaired) PEP005 treatment was associated with a >70% increase in tumor relapse rates. NK cell or monocyte/macrophage deficiency had no effect on relapse rates. Both in vitro and in mice, PEP005 induced MIP-2/IL-8, TNF-alpha, and IL-1beta, all mediators of neutrophil recruitment and activation. In vitro, PEP005 activated human endothelial cells resulting in neutrophil adhesion and also induced human neutrophils to generate tumoricidal-reactive oxygen intermediates. Treatment of tumors with PEP005 significantly elevated the level of anticancer Abs, which were able to promote neutrophil-mediated Ab-dependent cellular cytotoxicity (ADCC) in vitro. PEP005 treatment of tumors grown in SCID mice was also associated with >70% increase in tumor relapse rates. Taken together, these data suggest a central role for neutrophil-mediated ADCC in preventing relapse. PEP005-mediated cure of tumors therefore appears to involve initial chemoablation followed by a neutrophil dependent ADCC-mediated eradication of residual disease, illustrating that neutrophils can be induced to mediate important anticancer activity with specific chemotherapeutic agents. PMID- 17114488 TI - Recombinant soluble forms of extracellular TLR4 domain and MD-2 inhibit lipopolysaccharide binding on cell surface and dampen lipopolysaccharide-induced pulmonary inflammation in mice. AB - In this study, we sought the possibility of a new therapeutic strategy for dampening endotoxin-induced inflammation using soluble form of extracellular rTLR4 domain (sTLR4) and soluble form of rMD-2 (sMD-2). Addition of sTLR4 plus sMD-2 was significantly effective in inhibiting LPS-elicited IL-8 release from U937 cells and NF-kappaB activation in the cells transfected with TLR4 and MD-2 when compared with a single treatment with sTLR4 or sMD-2. Thus, we investigated the role of the extracellular TLR4 domain in interaction of lipid A with MD-2. Biotinylated sTLR4 failed to coprecipitate [(3)H]lipid A when it was sedimented with streptavidin-agarose, demonstrating that the extracellular TLR4 domain does not directly bind lipid A by itself. The amounts of lipid A coprecipitated with sMD-2 significantly increased when coincubated with sTLR4, and sTLR4 increased the affinity of lipid A for the binding to sMD-2. Soluble CD14 is required for the sTLR4-stimulated increase of lipid A binding to sMD-2. We also found that addition of sTLR4 plus sMD-2 inhibited the binding of Alexa-conjugated LPS to the cells expressing TLR4 and MD-2. Murine lungs that had received sTLR4 plus sMD-2 with LPS did not show any findings indicative of interstitial edema, neutrophil flux, and hemorrhage. Co-instillation of sTLR4 plus sMD-2, but not sTLR4 or sMD-2 alone, significantly decreased neutrophil infiltration and TNF-alpha levels in bronchoalveolar lavage fluids from LPS-treated mice. This study provides novel usage of sTLR4 and sMD-2 as an antagonist against endotoxin-induced pulmonary inflammation. PMID- 17114489 TI - Gene therapy of arthritis with TCR isolated from the inflamed paw. AB - In recent years, the treatment of autoimmune diseases has been significantly advanced by the use of biological agents. However, some biologics are accompanied with severe side effects, including tuberculosis and other types of infection. There is thus a critical need for nonsystemic and lesion-specific methods of delivering these therapeutic agents. We attempted to treat a mouse model of arthritis by using T cells that expressed a regulatory molecule and were specifically directed to the inflamed paw. To this end, we first identified the TCR alphabeta genes accumulating in the inflamed paw of mice with collagen induced arthritis (CIA) by a combination of single-strand chain polymorphism analysis of TCR and single-cell sorting. We identified an expanded clone B47 which is autoreactive but is not specific to type II collagen. In vivo, TCR genes from B47-transduced T cells accumulated in the inflamed paw. Injection of cells cotransduced with the B47 and soluble TNFRIg genes resulted in a significant suppression of CIA. The suppression was correlated with the amount of TNFRIg transcripts in the hind paw, not with the serum concentrations of TNFRIg. Moreover, T cells cotransduced with the B47 and intracellular Foxp3 genes significantly suppressed CIA with reductions in TNF-alpha, IL-17A, and IL-1beta expression and bone destruction. T cells cotransduced with B47 and Foxp3 genes also suppressed the progression of established CIA. Therefore, immunosuppressive therapy with autoreactive TCR is a promising therapeutic strategy for arthritis whether the TCRs are used to deliver either soluble or intracellular suppressive molecules. PMID- 17114490 TI - Annexin 1 negatively regulates IL-6 expression via effects on p38 MAPK and MAPK phosphatase-1. AB - Annexin 1 (Anx-1) is a mediator of the anti-inflammatory actions of glucocorticoids, but the mechanism of its anti-inflammatory effects is not known. We investigated the role of Anx-1 in the regulation of the proinflammatory cytokine, IL-6. Lung fibroblast cell lines derived from Anx-1(-/-) and wild-type (WT) mice were treated with dexamethasone and/or IL-1. IL-6 mRNA and protein were measured using real-time PCR and ELISA, and MAPK pathway activation was studied. Compared with WT cells, unstimulated Anx-1(-/-) cells exhibited dramatically increased basal IL-6 mRNA and protein expression. In concert with this result, Anx-1 deficiency was associated with increased basal phosphorylated p38, JNK, and ERK1/2 MAPKs. IL-1-inducible phosphorylated p38 was also increased in Anx-1(-/-) cells. The increase in IL-6 release in Anx-1(-/-) cells was inhibited by inhibition of p38 MAPK. Anx-1(-/-) cells were less sensitive to dexamethasone inhibition of IL-6 mRNA expression than WT cells, although inhibition by dexamethasone of IL-6 protein was similar. MAPK phosphatase-1 (MKP-1), a glucocorticoid-induced negative regulator of MAPK activation, was up-regulated by dexamethasone in WT cells, but this effect of dexamethasone was significantly impaired in Anx-1(-/-) cells. Treatment of Anx-1(-/-) cells with Anx-1 N-terminal peptide restored MKP-1 expression and inhibited p38 MAPK activity. These data demonstrate that Anx-1 is an endogenous inhibitory regulator of MAPK activation and IL-6 expression, and that Anx-1 is required for glucocorticoid up-regulation of MKP-1. Therapeutic manipulation of Anx-1 could provide glucocorticoid mimicking effects in inflammatory disease. PMID- 17114491 TI - The role of TLR2 in vivo following challenge with Staphylococcus aureus and prototypic ligands. AB - Based on a wealth of in vitro macrophage studies, immunity to Staphylococcus aureus cell wall-derived peptidoglycan (PGN) and lipoteichoic acid has been attributed to TLR2. We investigated whether the in vitro paradigm of TLR2 dominance would hold true in vivo. Using an experimental peritonitis model, we challenged mice with PGN or lipoteichoic acid and found that only PGN resulted in significant leukocyte (primarily neutrophil) accumulation in the peritoneum at 4 h. PGN-mediated leukocyte recruitment was P-/E-selectin dependent but only partially TLR2 dependent, and also involved the C5aR. Concomitant inhibition of TLR2 and C5aR resulted in a further reduction in PGN-induced peritonitis. Peritoneal neutrophilia was partially mast cell dependent; however, the defect could not be reconstituted with TLR2(-/-) or C5aR(-/-) mast cells. Interestingly, macrophage-deficient mice did not have defective neutrophil recruitment. By 24 h, the response to PGN involved primarily monocytes and was TLR2 and C5aR independent. Finally, we challenged mice with live S. aureus and found a similar degree of TLR2 involvement in leukocyte recruitment to that observed with PGN. Most importantly, bacterial clearance from the spleen and peritoneum was not altered in TLR2(-/-) mice vs wild-type mice. Morbidity was only significantly increased in S. aureus-infected mice treated with a blocking Fab against C5aR. Taken together, these studies indicate that in vivo responses to prototypic TLR2 ligands do not necessarily recapitulate the absolute necessity for TLR2 observed in vitro, and additional receptors contribute, in a significant manner, to PGN and S. aureus-mediated immune responses. PMID- 17114492 TI - Oligodeoxynucleotides differentially modulate activation of TLR7 and TLR8 by imidazoquinolines. AB - Among the 11 human TLRs, a subfamily TLR7, TLR8, and TLR9 display similarities in structure and endosomal localization. Natural agonists consisting of nucleic acids, such as ssRNA or DNA with CpG motifs, activate the innate immune cells through these TLRs. Immune response modifiers (IRMs) of imidazoquinoline class compounds 3M-001, 3M-002, and 3M-003 have been shown to activate the innate immune system via TLR7, TLR8, and TLR7/8, respectively. In looking at the effect of the agonists of the TLR7, TLR8, and TLR9 on the activation of NF-kappaB of transfected HEK cells, we discovered that some oligodeoxynucleotides (ODNs) could modulate imidazoquinoline effects in a negative or positive manner. In this study we demonstrate that poly(T) ODNs can inhibit TLR7 and enhance TLR8 signaling events involving NF-kappaB activation in HEK cells and cytokine production (IFN alpha, TNF, and IL-12) by human primary PBMC. In contrast, TLR3 agonist poly(I:C) does not affect imidazoquinoline-induced responses. The modulation of TLR7 and TLR8 responses is independent of CpG motifs or the nature of the ODN backbone structure. Furthermore, we show that to be an effective modulator, the ODNs need to be in the cell at the same time with either of the TLR7 or TLR8 agonist. We have also demonstrated that there is a physical interaction between IRMs and ODNs. The cross-talk between ODNs, IRMs, and TLR7 and TLR8 uncovered by this study may have practical implications in the field of microbial infections, vaccination, and tumor therapy. PMID- 17114493 TI - Plasminogen activator inhibitor type-1-deficient mice have an enhanced IFN-gamma response to lipopolysaccharide and staphylococcal enterotoxin B. AB - Plasminogen activator inhibitor type-1 (PAI-1) is a major inhibitor of fibrinolysis by virtue of its capacity to inhibit urokinase-type plasminogen activator (uPA) and tissue-type plasminogen activator (tPA). Systemic inflammation is invariably associated with elevated circulating levels of PAI-1, and during human sepsis plasma PAI-1 concentrations predict an unfavorable outcome. Knowledge about the functional role of PAI-1 in a systemic inflammatory response syndrome is highly limited. In this study, we determined the role of endogenous PAI-1 in cytokine release induced by administration of LPS or staphylococcal enterotoxin B (SEB). Both LPS and SEB elicited secretion of PAI-1 into the circulation of normal wild-type (Wt) mice. Relative to Wt mice, PAI-1 gene-deficient (PAI-1(-/-)) mice demonstrated strongly elevated plasma IFN-gamma concentrations after injection of either LPS or SEB. In addition, PAI-1(-/-) splenocytes released more IFN-gamma after incubation with LPS or SEB than Wt splenocytes. Both PAI-1(-/-) CD4+ and CD8+ T cells produced more IFN-gamma upon stimulation with SEB. LPS-induced IFN-gamma release in mice deficient for uPA, the uPA receptor, or tPA was not different from IFN-gamma release in LPS-treated Wt mice. These results identify a novel function of PAI-1 during systemic inflammation, where endogenous PAI-1 serves to inhibit IFN-gamma release by a mechanism that does not depend on its interaction with uPA/uPA receptor or tPA. PMID- 17114494 TI - Selective localization of recognition complexes for leukotriene B4 and formyl-Met Leu-Phe within lipid raft microdomains of human polymorphonuclear neutrophils. AB - Neutrophilic polymorphonuclear leukocytes contain glycosphingolipid- and cholesterol-enriched lipid raft microdomains within the plasma membrane. Although there is evidence that lipid rafts function as signaling platforms for CXCR chemokine receptors, their role in recognition systems for other chemotaxins such as leukotriene B4 (LTB4) and fMLP is unknown. To address this question, human neutrophils were extracted with 1% Brij-58 and fractionated on sucrose gradients. B leukotriene receptor-1 (BLT-1), the primary LTB4 receptor, partitioned to low density fractions, co-isolating with the lipid raft marker, flotillin-1. By contrast, formyl peptide receptor (FPR), the primary fMLP receptor, partitioned to high density fractions, co-isolating with a non-raft marker, Cdc42. This pattern was preserved after the cells were stimulated with LTB4 or fMLP. Fluorescence resonance energy transfer (FRET) was performed to confirm the proximity of BLT-1 and FPR with these markers. FRET was detected between BLT1 and flotillin-1 but not Cdc42, whereas FRET was detected between FPR and Cdc42, but not flotillin-1. Pretreating neutrophils with methyl-beta-cyclodextrin, a lipid raft-disrupting agent, suppressed intracellular Ca(2+) mobilization and ERK1/2 phosphorylation in response to LTB4 but had no effect on either of these responses to fMLP. We conclude that BLT-1 is physically located within lipid raft microdomains of human neutrophils and that disrupting lipid raft integrity suppresses LTB4-induced activation. By contrast, FPR is not associated with lipid rafts, and fMLP-induced signaling does not require lipid raft integrity. These findings highlight the complexity of chemotaxin signaling pathways and offer one mechanism by which neutrophils may spatially organize chemotaxin signaling within the plasma membrane. PMID- 17114495 TI - Antigen-specific central memory CD4+ T lymphocytes produce multiple cytokines and proliferate in vivo in humans. AB - The function of Ag-specific central (T(CM)) and effector (T(EM)) memory CD4+ T lymphocytes remains poorly characterized in vivo in humans. Using CD154 as a marker of Ag-specific CD4+T cells, we studied the differentiation of memory subsets following anti-hepatitis B immunization. Hepatitis B surface Ag (HBs) specific memory CD4+T cells were heterogeneous and included T(CM) (CCR7+CD27+) and T(EM) (CCR7(-)CD27(+/-)). HBs-specific T(CM) and T(EM) shared the capacity to produce multiple cytokines, including IL-2 and IFN-gamma. Several years postimmunization, approximately 10% of HBs-specific memory CD4+ T cells were in cycle (Ki67+) and the proliferating cells were CCR7+. These results suggest that the model of functional specialization of T(CM) and T(EM) cannot be applied to protein vaccine Ags and support the concept that T(CM) are capable of self renewal and contribute to maintain the pool of memory cells. PMID- 17114496 TI - CD8-mediated type 1 antitumor responses selectively modulate endogenous differentiated and nondifferentiated T cell localization, activation, and function in progressive breast cancer. AB - CD8 T cell-mediated immune responses fall into two distinct types based on effector cell-derived cytokine production. Type I CD8 T cells (Tc1) produce IFN gamma, whereas type 2 cells (Tc2) secrete IL-4, IL-5, IL-10, and GM-CSF. Using a murine TCR transgenic T cell/breast tumor model, we show that adoptively transferred Ag-specific Tc1 cells are more effective in delaying mammary tumor growth and progression than that of functionally distinct Tc2 cells. Donor Tc1 cells administered 7 days posttumor challenge localized and persisted at sites of primary tumor growth with antitumor responses that were dependent, in part, on effector cell-derived IFN-gamma. Tc1-mediated responses markedly enhanced the appearance and local accumulation of highly differentiated (CD44(high)) CD4 and CD8 endogenous tumor-infiltrating T cells when compared with that of untreated tumor-bearing mice. Conversely, Tc1 cell transfer markedly delayed the appearance of corresponding nondifferentiated (CD44(low)) endogenous T cells. Such cells were acutely activated as defined by coexpression of surface markers associated with TCR engagement (CD69) and early T cell activation (CD25). Moreover, cellular response kinetics appeared to further correlate with the up-regulation of endogenous T cells producing the chemokine IFN-gamma-inducible protein-10 in vivo. This suggested that CD8-mediated type 1 antitumor responses cannot only promote accumulation of distinct endogenous CD4 and CD8 T cell subpopulations, but also facilitate and preferentially modulate their localization kinetics, persistence, states of activation/differentiation, and function within the primary tumor environment at various stages of tumor progression. These studies offer insight into potential mechanisms for enhancing T cell-based immunotherapy in breast cancer. PMID- 17114497 TI - IRAK-4 mutation (Q293X): rapid detection and characterization of defective post transcriptional TLR/IL-1R responses in human myeloid and non-myeloid cells. AB - Innate immunodeficiency has recently been reported as resulting from the Q293X IRAK-4 mutation with consequent defective TLR/IL-1R signaling. In this study we report a method for the rapid allele-specific detection of this mutation and demonstrate both cell type specificity and ligand specificity in defective IL-1R associated kinase (IRAK)-4-deficient cellular responses, indicating differential roles for this protein in human PBMCs and primary dermal fibroblasts and in LPS, IL-1beta, and TNF-alpha signaling. We demonstrate transcriptional and post transcriptional defects despite NF-kappaB signaling and intact MyD88-independent signaling and propose that dysfunctional complex 1 (IRAK1/TRAF6/TAK1) signaling, as a consequence of IRAK-4 deficiency, generates specific defects in MAPK activation that could underpin this patient's innate immunodeficiency. These studies demonstrate the importance of studying primary human cells bearing a clinically relevant mutation; they underscore the complexity of innate immune signaling and illuminate novel roles for IRAK-4 and the fundamental importance of accessory proinflammatory signaling to normal human innate immune responses and immunodeficiencies. PMID- 17114498 TI - Spontaneous CD8 T cell responses against the melanocyte differentiation antigen RAB38/NY-MEL-1 in melanoma patients. AB - The melanocyte differentiation Ag RAB38/NY-MEL-1 was identified by serological expression cloning (SEREX) and is expressed in the vast majority of melanoma lesions. The immunogenicity of RAB38/NY-MEL-1 has been corroborated previously by the frequent occurrence of specific Ab responses in melanoma patients. To elucidate potential CD8 T cell responses, we applied in vitro sensitization with overlapping peptides spanning the RAB38/NY-MEL-1 protein sequence and the reverse immunology approach. The identified peptide RAB38/NY-MEL-1(50-58) exhibited a marked response in ELISPOT assays after in vitro sensitization of CD8 T cells from HLA-A *0201(+) melanoma patients. In vitro digestion assays using purified proteasomes provided evidence of natural processing of RAB38/NY-MEL-1(50-58) peptide. Accordingly, monoclonal RAB38/NY-MEL-1(50-58)-specific T cell populations were capable of specifically recognizing HLA-A2(+) melanoma cell lines expressing RAB38/NY-MEL-1. Applying fluorescent HLA-A2/RAB38/NY-MEL-1(50 58) multimeric constructs, we were able to document a spontaneously developed memory/effector CD8 T cell response against this peptide in a melanoma patient. To elucidate the Ag-processing pathway, we demonstrate that RAB38/NY-MEL-1(50-58) is produced efficiently by the standard proteasome and the immunoproteasome. In addition to the identification of a RAB38/NY-MEL-1-derived immunogenic CD8 T cell epitope, this study is instrumental for both the onset and monitoring of future RAB38/NY-MEL-1-based vaccination trials. PMID- 17114499 TI - Antibodies against the activated coagulation factor X (FXa) in the antiphospholipid syndrome that interfere with the FXa inactivation by antithrombin. AB - Antiphospholipid Ab have been shown to promote thrombosis and fetal loss in the antiphospholipid syndrome (APS). Previously, we found IgG anti-thrombin Ab in some APS patients that could interfere with inactivation of thrombin by antithrombin (AT). Considering that activated coagulation factor X (FXa) is homologous to thrombin in the catalytic domains and is also regulated primarily by AT, we hypothesized that some thrombin-reactive Ab may bind to FXa and interfere with AT inactivation of FXa. To test these hypotheses, we studied reactivity of eight patient-derived monoclonal IgG antiphospholipid Ab with FXa and the presence of IgG anti-FXa Ab in APS patients and investigated the effects of FXa-reactive mAb on AT inactivation of FXa. The results revealed that six of six thrombin-reactive IgG mAb bound to FXa and that the levels of plasma IgG anti FXa Ab in 38 APS patients were significantly higher than those in 30 normal controls (p < 0.001). When the mean plus 3 SDs of the 30 normal controls was used as the cutoff, 5 of 38 APS patients (13.2%) had IgG anti-FXa Ab. Importantly, three of six FXa-reactive mAb significantly inhibited AT inactivation of FXa. Combined, these results indicate that anti-FXa Ab may contribute to thrombosis by interfering with the anticoagulant function of AT on FXa in some APS patients. PMID- 17114500 TI - Synovial autoreactive T cells in rheumatoid arthritis resist IDO-mediated inhibition. AB - A hallmark of T cell-mediated autoimmunity is the persistence of autoreactive T cells. However, it remains to elucidate the manner in which synovial T cells are sustained in patients with rheumatoid arthritis (RA). We found that dendritic cells (DC) and tissues from the synovial joints of RA patients expressed higher levels of IDO than DC from healthy donors. Interestingly, T cells derived from the joint synovial fluid (SF) of RA patients proliferated in response to either autologous or allogeneic IDO-positive DC, an outcome that was not affected by the addition of IDO inhibitor 1-methyl-D-tryptophan (1-MT). In contrast, addition of 1-MT to the culture stimulated with allogeneic or autologous IDO-positive DC significantly enhanced the proliferation of T cells derived from peripheral blood of healthy donors or from peripheral blood of RA patients. Furthermore, we found that functionally active tryptophanyl-tRNA-synthetase (TTS) was significantly elevated in T cells derived from the SF of RA patients, leading to enhanced storage of tryptophan in T cells and to subsequent resistance to IDO-mediated deprivation of tryptophan. The RA SF enhancement of TTS expression in T cells was blocked by mAb to IFN-gamma and TNF-alpha. These results suggest that the resistance of T cells to IDO-mediated deprivation of tryptophan represents a mechanism by which autoreactive T cells are sustained in vivo in RA patients. Specifically, blocking of the up-regulation of TTS expression in T cells presents an avenue for development of a novel therapeutic approach to treatment of RA. PMID- 17114501 TI - Mimicry and antibody-mediated cell signaling in autoimmune myocarditis. AB - The mechanisms by which autoantibodies against cardiac myosin (CM) may lead to heart dysfunction is unknown. We show that autoantibodies to CM in anti-CM sera and mAbs derived from experimental autoimmune myocarditis targeted the heart cell surface and induced Ab-mediated cAMP-dependent protein kinase A activity. Ab mediated cell signaling of protein kinase A was blocked by CM, anti-IgG, or by specific inhibitors of the beta-adrenergic receptor (beta-AR) pathway. mAbs confirmed mimicry between CM and the beta-AR. Passive transfer of purified Ab (IgG) from CM-immunized rats resulted in IgG deposition and apoptosis in the heart, leading to a cardiomyopathic heart disease phenotype in recipients. Our novel findings link anti-CM Ab with the beta-AR and subsequent Ab-mediated cell signaling in the heart. PMID- 17114502 TI - Coadministration of plasmid DNA constructs encoding an encephalitogenic determinant and IL-10 elicits regulatory T cell-mediated protective immunity in the central nervous system. AB - We have previously shown that Ag-specific IL-10-producing regulatory T cells (Tr1) participate in the regulation of experimental autoimmune encephalomyelitis and that their specificity undergoes determinant spread in a reciprocal manner to effector T cell specificity. The current study shows that coadministration of plasmid DNA vaccines encoding IL-10 together with a plasmid encoding a myelin basic protein (MBP) encephalitogenic determinant during an ongoing disease rapidly amplifies this Tr1-mediated response, in a disease-specific manner. Thus, coadministration of both plasmids, but not the plasmid DNA encoding MBP alone, rapidly suppresses an ongoing disease. Tolerance included elevation in Ag specific T cells producing IL-10 and an increase in apoptosis of cells around high endothelial venules in the CNS after successful therapy. Tolerance could be transferred by MBP-specific primary T cells isolated from protected donors and reversed by neutralizing Abs to IL-10 but not to IL-4. Due to the nature of determinant spread in this model, we could bring about evidence implying that rapid and effective induction of Tr1-induced active tolerance is dependent on redirecting the Tr1 response to the epitope to which the effector function dominates the response at a given time. The consequences of these findings to multiple sclerosis, and possibly other inflammatory autoimmune diseases are discussed. PMID- 17114503 TI - Stability and translation of TCR zeta mRNA are regulated by the adenosine-uridine rich elements in splice-deleted 3' untranslated region of zeta-chain. AB - Systemic lupus erythematosus (SLE) T cells display reduced expression of TCR zeta protein. Recently, we reported that in SLE T cells, the residual TCR zeta protein is predominantly derived from an alternatively spliced form that undergoes splice deletion of 562 nt (from 672 to 1233 bases) within the 3' untranslated region (UTR) of TCR zeta mRNA. The stability and translation of the alternatively spliced form of TCR zeta mRNA are low compared with that of the wild-type TCR zeta mRNA. We report that two adenosine-uridine-rich sequence elements (AREs), defined by the splice-deleted 3' UTR region, but not an ARE located upstream are responsible for securing TCR zeta mRNA stability and translation. The stabilizing effect of the splice-deleted region-defined AREs extended to the luciferase mRNA and was not cell type-specific. The findings demonstrate distinct sequences within the splice-deleted region 672 to 1233 of the 3' UTR, which regulate the transcription, mRNA stability, and translation of TCR zeta mRNA. The absence of these sequences represents a molecular mechanism that contributes to altered TCR zeta-chain expression in lupus. PMID- 17114504 TI - Role for nephritogenic T cells in lupus glomerulonephritis: progression to renal failure is accompanied by T cell activation and expansion in regional lymph nodes. AB - Autoreactive T cells are critical in the initiation and maintenance of autoantibody responses that are a hallmark of systemic lupus erythematosus. However, the direct contribution of T cells in end-organ disease like lupus glomerulonephritis (GN) is poorly understood. In this study, we investigated the role of T cells in progression of lupus GN in NZM2328 mice, a murine model of spontaneous systemic lupus erythematosus. At 26 wk of age, NZM2328 female mice showed glomerular immune complex deposits and acute proliferative GN. This was associated with up-regulation of MHC class II and the detection of T cells and CD11c(+) dendritic cells in the glomeruli. The regional lymph nodes (LN) showed preferential activation of T cells and an oligoclonal T cell response with skewed expansion of certain Vbeta families. This suggests an Ag-driven response occurring in the regional LN of nephritic mice during acute GN. In contrast, male NZM2328 mice developed glomerular immune complexes and acute GN, but rarely progressed to fatal chronic GN. Significantly, male kidneys at 40 wk of age did not have detectable dendritic cells and T cells in the glomeruli. Thus, glomerular immune complex deposition initiates an immune response against renal Ags in the regional LN, leading to T cell recruitment into the kidney during acute proliferative GN. This T cell activation and infiltration are influenced by gender-dependent end-organ factors and may determine the progression of acute GN to chronic GN and renal failure. PMID- 17114505 TI - IL-22-mediated tumor growth reduction correlates with inhibition of ERK1/2 and AKT phosphorylation and induction of cell cycle arrest in the G2-M phase. AB - IL-22 is a recently discovered cytokine of the IL-10 family that binds to a class II cytokine receptor composed of IL-22R1 and IL-10R2(c) and influences a variety of immune reactions. As IL-22 has also been shown to modulate cell cycle and proliferation mediators such as ERK1/2 and JNK, we studied the role of IL-22 in proliferation, apoptosis, and cell cycle regulation in EMT6 murine breast cancer cells in vitro and in vivo. In this study, we report that murine breast cancer cells express functional IL-22R as indicated by RT-PCR studies, immunoblotting, and STAT3 activation assays. Importantly, IL-22 exposure of EMT6 cells resulted in decreased levels of phosphorylated ERK1/2 and AKT protein kinases, indicating an inhibitory effect of IL-22 on signaling pathways promoting cell proliferation. Furthermore, IL-22 induced a cell cycle arrest of EMT6 cells in the G(2)-M phase. IL-22 reduced EMT6 cell numbers and the proliferation rate by approximately 50% as measured by [(3)H]thymidine incorporation. IL-22 treatment of EMT6 tumor bearing mice lead to a decreased tumor size and a reduced tumor cell proliferation in vivo, as determined by 3'-deoxy-3'-fluorothymidine-positron emission tomography scans. Interestingly, IL-22 did not induce apoptosis, as determined in annexin V binding assay and caspase-3 activation assay and had no effect on angiogenesis in vivo. In conclusion, our results indicate that IL-22 reduced tumor growth by inhibiting signaling pathways such as ERK1/2 and AKT phosphorylation that promote tumor cell proliferation in EMT6 cells. Therefore, IL-22 may play a role in the control of tumor growth and tumor progression. PMID- 17114506 TI - Intensity of signal contacting meniscal surface in recurrent tears on MR arthrography compared with that of contrast material. AB - OBJECTIVE: Several previous studies reported that the signal contacting the meniscal surface in a recurrent tear on MR arthrography had intensity equal to that of intraarticular contrast material. Because we failed to diagnose recurrent tears using this criterion, we reviewed our knee MR arthrograms in patients who had prior meniscal surgery. CONCLUSION: On knee MR arthrograms, the signal contacting the surface of a recurrent meniscal tear may be equal to or less than that of adjacent intraarticular gadolinium contrast material. PMID- 17114507 TI - MRI of Takayasu's arteritis: typical appearances and complications. AB - OBJECTIVE: Although Takayasu's arteritis (TA) is more common in Asian countries, it has a worldwide distribution. MRI is widely used for its diagnosis. Our purpose is to present a comprehensive pictorial review of its typical appearances and complications on MRI. CONCLUSION: MRI can effectively provide almost all anatomic information in patients with TA. MRI is very useful for accurate diagnosis of TA and its complications. Knowledge of MRI findings is essential for improving patient outcome. PMID- 17114508 TI - Comparison of full-field digital mammography and screen-film mammography for detection and characterization of simulated small masses. AB - OBJECTIVE: The two objectives of this study were to create an ex vivo phantom model that closely mimics human breast cancer for detection tasks and to compare the performance of full-field digital mammography with screen-film mammography in detecting and characterizing small breast masses in a phantom with a spectrum of complex tissue backgrounds. MATERIALS AND METHODS: Sixteen phantom breast masses of varying sizes (0.3-1.2 cm), shapes (round and irregular), and densities (high and low) were created from shaved tumor specimens and imaged using both full field digital and screen-film mammography techniques. We created 408 detection tasks that were captured on 68 films. On each radiograph, six detection tasks were partially obscured by areas of varying breast-pattern complexity, including low (predominantly fatty), mixed (scattered fibroglandular densities and heterogeneously dense), and high (extremely dense) density patterns. Each detection task was scored using a five-point confidence scale by three mammographers. Receiver operating characteristic (ROC) curve analysis was performed to analyze differences in detection of masses between the two imaging systems, and sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were computed. RESULTS: Full-field digital mammography showed higher area under the ROC curve than screen-film mammography for detecting masses in each breast background and performed significantly better than screen-film mammography in mixed (p = 0.010), dense (p = 0.029), and all breast backgrounds combined (p = 0.004). Full-field digital mammography was superior to screen-film mammography for characterizing round and irregular masses and low- and high-density masses. CONCLUSION: Full-field digital mammography was significantly superior to screen-film technique for detecting and characterizing small masses in mixed and dense breast backgrounds in a phantom model. PMID- 17114509 TI - Comparison of lymphotropic nanoparticle-enhanced MRI sequences in patients with various primary cancers. AB - OBJECTIVE: This study was performed to empirically evaluate T2-weighted fast spin echo, moderately T2*-weighted gradient-refocused echo (GRE), and heavily T2* weighted GRE sequences to determine which sequence is the most effective for nodal characterization on lymphotropic nanoparticle-enhanced MRI (LNMRI). MATERIALS AND METHODS: The study included 65 patients who had proven primary cancer and were scheduled for either surgical lymph node dissection or imaging guided lymph node biopsy. All patients underwent LNMRI using T2-weighted fast spin-echo, moderately T2*-weighted GRE, and heavily T2*-weighted GRE sequences. Unequivocal correlation of histopathology and MRI could be made in 140 nodes and only these were included in the analysis. Two blinded reviewers performed qualitative analysis of the nodes. Alternative free-response receiver operating characteristic (ROC) curves with a continuous rating scale were plotted for each sequence for both reviewers and the diagnostic accuracy of fast spin-echo T2 weighted and GRE T2*-weighted images were compared by calculating the area under the curve (A(Z)). A two-tailed Student's t test was performed to test the significance (p < 0.05) of the differences between the ROC curves derived from the three sequences. RESULTS: Irrespective of reviewer experience, T2*-weighted sequences showed better nodal characterization when compared with T2-weighted sequences. For both reviewers, there was a statistically significant difference between the A(Z) for T2- and the two T2*-weighted sequences (p < 0.05). Neither reviewer showed a statistically significant difference between the two T2* weighted sequences. CONCLUSION: GRE T2*-weighted sequences are superior for nodal characterization on LNMRI to fast spin-echo T2-weighted sequences. Imaging protocols for LNMRI should include fast spin-echo T2-weighted imaging for anatomic localization, but characterization of nodes should be based on their appearance on contrast-enhanced T2*-weighted images. The T2*-weighted images acquired with dual TE values, one of which is intermediate and the other longer, improve nodal characterization. PMID- 17114510 TI - Prevalence and sonographic detection of Chilaiditi's sign in cirrhotic patients without ascites. AB - OBJECTIVE: Although Chilaiditi's sign is uncommon, its recognition is mandatory to avoid intestinal injury during percutaneous transhepatic procedures. Our purpose was to investigate the prevalence of Chilaiditi's sign in cirrhotic patients without ascites and to review the diagnostic ability of sonography to detect this rare abnormality. CONCLUSION: The prevalence of Chilaiditi's sign was much higher in cirrhotic patients without ascites than in the general population. A precise diagnosis of Chilaiditi's sign was possible using sonography. Percutaneous transhepatic procedures can be performed safely if a route that avoids the intestine is found. PMID- 17114511 TI - Pouchography, CT, and MRI features of ileal J pouch-anal anastomosis. AB - OBJECTIVE: Our objective is to describe pouchography, CT, and MRI features of the J-shaped pouch, both normal and with pouch-related complications. CONCLUSION: Pouchography is performed before closure of the loop ileostomy to assess the integrity of the ileal pouch and anastomosis. CT and MRI can be performed when postoperative complications, such as small-bowel obstruction, pouchitis, leakage, abscess, intramural hematoma, desmoid tumor, or recurrent Crohn's disease, are suspected. PMID- 17114512 TI - Intraductal papillary mucinous tumors of the pancreas: branch duct tumor penetrating the stomach and duodenum. PMID- 17114513 TI - Heterotopic pancreas: presentation as jejunal tumor. PMID- 17114514 TI - 99mTc-MAG3 renography: normal values for MAG3 clearance and curve parameters, excretory parameters, and residual urine volume. AB - OBJECTIVE: Specific quantitative measurements have been recommended to assist in the interpretation of technetium-99m mercaptoacetyltriglycine (MAG3) renal studies. Our objective was to define the sex- and age-specific normal ranges for these recommended parameters. MATERIALS AND METHODS: Data were obtained from a retrospective analysis of 106 subjects who were evaluated for kidney donation. The MAG3 clearance was calculated using a common camera-based method. The relative uptake, prevoid/postvoid and postvoid/maximum count ratios were determined using whole-kidney regions of interest (ROIs). Time to peak, time to half-peak, 20 min/maximum and 20 min/2-3 min count ratios were determined for cortical and whole-kidney ROIs. Residual urine volume was calculated on the basis of the pre- and postvoid bladder counts and voided urine volume. RESULTS: The mean camera-based MAG3 clearance was 321 +/- 69 mL/min/1.73 m2, essentially the same as the mean plasma sample MAG3 clearance in comparable populations. The percentages of relative uptake in the right and left kidneys were 49% and 51% +/- 4%, respectively; no difference was seen between men and women. Cortical values were lower than the whole-kidney values (p < 0.001); the mean cortical 20 min/maximum count ratio was 0.19 (SD, 0.07 and 0.04 for right and left kidneys, respectively). The mean postvoid/maximum whole-kidney count ratio was < 0.1, and the mean postvoid residual bladder volume was < 30 mL. CONCLUSION: Normal limits adjusted for age and sex have been established. Applying normal ranges to quantitative MAG3 parameters may assist in the interpretation of MAG3 scintigraphy and facilitate appropriate patient management. PMID- 17114515 TI - 18F-choline PET/CT for initial staging of advanced prostate cancer. PMID- 17114516 TI - Hepatic visceral larva migrans of Toxocara canis: CT and sonographic findings. AB - OBJECTIVE: The aim of this study was to describe the CT and sonographic findings of hepatic visceral larva migrans of Toxocara canis. MATERIALS AND METHODS: Fifty four patients (44 men, 10 women; age range, 30-80 years; mean age, 53 years) with serologically confirmed visceral larva migrans of Toxocara canis underwent evaluation of the liver with CT (n = 25), sonography (n = 48), or both. Two radiologists used consensus for retrospective evaluation of CT and sonographic findings. Correlation between the presence and severity of hepatic abnormalities on images and the degree of peripheral eosinophilia was assessed. RESULTS: Seventeen (68%) of 25 patients who underwent CT had single or multiple ill defined, oval or elongated, small, low-attenuating lesions in the liver. Eighteen (38%) of 48 patients who underwent sonography had single or multiple small, poorly defined, oval or elongated, hypoechoic scattered focal lesions in the liver. In the 19 patients who underwent both CT and sonography, the two techniques had no significant difference in rate of detection of hepatic lesions (p = 0.375, McNemar test). The lesion numbers on CT and sonography showed excellent linear correlation (r = 0.844, p = 0.001) by Pearson's correlation test. An independent samples t test showed that eosinophil count and percentage in the peripheral blood were significantly higher in patients with hepatic lesions on CT and sonography than in patients without lesions. CONCLUSION: CT and sonographic findings of hepatic visceral larva migrans of T. canis are multiple, ill-defined, oval or elongated, small, nodular lesions scattered in the liver parenchyma. The presence of hepatic lesions on images was associated with higher peripheral eosinophil count and percentage. PMID- 17114517 TI - Myocardial T1 mapping for detection of left ventricular myocardial fibrosis in chronic aortic regurgitation: pilot study. AB - OBJECTIVE: The aim of this study was to identify diffuse myocardial fibrosis secondary to chronic aortic regurgitation by comparing the T1 relaxation times of left ventricular myocardium in a pilot patient group with a previously established normal range of times. SUBJECTS AND METHODS: Eight patients with chronic aortic regurgitation and normal coronary arteries awaiting surgical valve replacement underwent a comprehensive MRI examination that included assessment of left ventricular function, severity of valvular regurgitation, and presence of overt myocardial scar evidenced by delayed enhancement. For each patient, myocardial T1 relaxation times determined with a modified Look-Locker technique before and after contrast administration were compared with values previously established for 15 healthy volunteers. RESULTS: There was no statistical difference (p > 0.05) in slice-averaged myocardial T1 relaxation times either before or after gadolinium administration in the patient group compared with the normal range of times. Segmental averaged T1 relaxation times in segments with abnormal wall motion did, however, show statistically significant differences from healthy controls 10, 15, and 20 minutes after administration of gadolinium (510 vs 476 milliseconds, p = 0.001; 532 vs 501 milliseconds, p = 0.002; 560 vs 516 milliseconds, p = 0.001, respectively). Two of the aortic regurgitation patients also had focal areas of myocardial delayed enhancement. CONCLUSION: Segment-based myocardial T1 mapping has the potential for showing differences between relaxation times in aortic regurgitation and in normal hearts, suggesting the existence of a diffuse myocardial fibrotic process. PMID- 17114518 TI - Cytomegalovirus pneumonia after stem cell transplantation: correlation of CT findings with clinical outcome in 30 patients. AB - OBJECTIVE: The purpose of our study was to assess the correlation between early high-resolution CT findings of cytomegalovirus (CMV) pneumonia in patients with blood disorders and their clinical outcomes. CONCLUSION: The initial high resolution CT findings in immunocompromised patients with CMV pneumonia seem to predict the patient's outcome being unfavorable in those forms of disease beginning mostly bilaterally as diffuse or patchy ground-glass opacity followed by progressive air-space consolidation. Also, a change in the CT morphology of pulmonary lesions toward diffuse ground-glass opacity seems to correlate with an unfavorable disease course. PMID- 17114519 TI - Sonographically guided percutaneous liver biopsy in infants: a retrospective review. AB - OBJECTIVE: The purpose of our study was to assess the technical success and complication rate of sonographically guided percutaneous liver biopsies performed in infants under 1 year old at a tertiary pediatric center. MATERIALS AND METHODS: Retrospective analysis of 65 biopsies performed in 61 infants between January 1999 and December 2003 was conducted. Data collected included patient demographics; details of the biopsy procedure including indication, needle type and size, number of passes, and samples; pathology results; and procedure-related complications. RESULTS: The 61 infants studied included 37 males and 24 females with a mean age of 119 days (age range, 7-348 days; median age, 83 days) and a mean weight of 4.5 kg (1.9-8.3 kg). A total of 65 biopsies were performed in these 61 infants. General anesthesia was used in 66.1% of procedures. An 18-gauge needle was used in 47 (72.3%) procedures. Coaxial technique was used in seven procedures, and five biopsy tracts were embolized. In 63 of 65 procedures, the mean number of passes was 1.8. In two procedures, using a coaxial technique, 11 and 12 passes were made. One biopsy was considered technically unsuccessful, and 64 of 65 (98.5%) of the biopsies provided adequate tissue for pathologic analysis. There were three (4.6%) major complications related to bleeding: one requiring a blood transfusion, one requiring surgery, and one arteriobiliary fistula requiring transarterial embolization. Three (4.6%) minor complications also occurred. There were no deaths. CONCLUSION: Sonographically guided percutaneous liver biopsy in infants is a good and effective diagnostic tool. The complication rate, however, even when performed by an experienced physician, is not insignificant in this age group of patients. PMID- 17114520 TI - Progression of middle cerebral artery susceptibility sign on T2*-weighted images: its effect on recanalization and clinical outcome after thrombolysis. AB - OBJECTIVE: The middle cerebral artery (MCA) "susceptibility sign" on T2*-weighted imaging has been reported to indicate acute thrombotic occlusion. We evaluated the serial progression of this susceptibility sign on follow-up MRI and its effect on recanalization and clinical outcome after intraarterial thrombolysis. MATERIALS AND METHODS: Thirty-three acute ischemic stroke patients who were treated with intraarterial thrombolysis and underwent MRI within 6 hours of symptom onset were enrolled in this study. All study participants had either M1 or M2 occlusion on digital subtraction angiography before thrombolysis and underwent follow-up MRI 2-3 days after thrombolysis. Recanalization status was evaluated using the thrombolysis in myocardial infarction (TIMI) flow grade on digital subtraction angiography immediately after thrombolysis. The serial progression of the susceptibility sign on follow-up T2*-weighted imaging was compared with the MR angiographic findings. Baseline clinical parameters and clinical outcome were also reviewed. RESULTS: A positive MCA susceptibility sign on the initial T2*-weighted imaging was detected in 16 (48%) of the 33 patients. The mean TIMI grade was higher in the patients with a positive sign on imaging than in those without the sign (2.3 vs 1.0, respectively; p < 0.005). In the risk factor analysis, a history of atrial fibrillation was significantly higher in the patients with the MCA susceptibility sign than in those with negative findings for the sign (13/16 [81%] vs 4/17 [24%], respectively). In 14 of the 16 patients with the positive sign, the sign disappeared on follow-up MRI, and that finding (i.e., disappearance of the sign) was well correlated with complete recanalization on follow-up MR angiography in 12 patients. Multivariate logistic regression analysis showed that this sign was not associated with a favorable functional outcome 30 days after thrombolytic treatment. CONCLUSION: The MCA susceptibility sign can be indicative of acute thromboembolic occlusion and can be used to predict the immediate effectiveness of intraarterial thrombolysis. However, the appearance of this sign was not associated with a favorable clinical outcome after thrombolysis in our small series study. PMID- 17114521 TI - MR features of vertebral tophaceous gout. PMID- 17114522 TI - Postpartum spontaneous coronary artery dissection: a case of pseudoaneurysm evolution detected on MDCT. PMID- 17114523 TI - Utilization of the double-contrast barium enema in the early era of screening CT colonography. PMID- 17114524 TI - Incidence of myocardial bridging observed on MDCT. PMID- 17114525 TI - Imagining imaging: radiology practice in 2050. PMID- 17114526 TI - Development and management of a noninvasive cardiovascular imaging service. PMID- 17114527 TI - "Multiple fractures in the long bones of infants suffering from chronic subdural hematoma"--a commentary. PMID- 17114528 TI - Retirement patterns and plans of radiologists. AB - OBJECTIVE: The objectives of our study were to describe radiologists' recent retirement plans and patterns and to assess whether changes in radiologists' retirement patterns over the period of 1995-2003 explain the recent easing of the radiologist shortage. MATERIALS AND METHODS: We present detailed information from 2003 about the planned retirement age of radiologists, their labor force participation late in their careers, and their actual retirement pattern based on data from the American College of Radiology's (ACR) 2003 Survey of Radiologists. To analyze changes over time, we compare these data with information from the ACR's 1995 and 2000 Surveys of Radiologists. Multivariate regression analysis was also used to identify the effects of radiologist and practice characteristics on radiologists' retirement plans. RESULTS: The percentage of radiologists fully retired and the average retirement age were the same in 1995 and 2003. Overall, labor force participation rates were decreasing over the period 1995-2003 for both women and men. Standardized labor force participation rates for radiologists age 55-74 years appeared to decrease from 1995 to 2000 and remained at a lower level in 2003, but the changes were not statistically significant. As of 2003, radiologists retired at 64, approximately 2 years older than the average U.S. worker. CONCLUSION: Radiologists remain active in their profession longer than the typical U.S. worker. There was no change in radiologists' pattern of gradually moving into retirement. If anything, radiologists were retiring earlier in 2003 than in the past. A delay in retirement is not an explanation of the recent easing of the radiologist shortage. PMID- 17114529 TI - MRI findings of femoroacetabular impingement. AB - OBJECTIVE: The purpose of our study was to evaluate MRI in the identification of labral and articular cartilage lesions in patients with a clinical suspicion of femoroacetabular impingement. MATERIALS AND METHODS: Preoperative MRI was performed in 46 consecutive patients (26 men, 20 women; age range, 21-45 years; mean age, 32.3 years) for whom femoroacetabular impingement was clinically suspected. Two musculoskeletal radiologists independently assessed the MR images for the presence and anatomic site of labral disorders, labral-chondral transitional zone disorders, femoral cartilage lesions, and acetabular cartilage lesions. Surgical correlation was obtained in all cases by two surgeons who were experienced in hip arthroscopy. RESULTS: Seven patients showed labral tears on MRI that were confirmed surgically in all cases. Thirty-seven patients (97%) of the 38 surgically confirmed cases had lesions of the labral-chondral transitional zone on MRI. The sites of labral-chondral transitional zone abnormalities at arthroscopy were 50% anterosuperior, 36% anterosuperior and superolateral, 11% superolateral, and 3% superolateral and posterosuperior. The site was identified correctly in 92% (reviewer 1) and 95% (reviewer 2) of cases on MRI. Separate acetabular cartilage abnormality was surgically identified in 39% of cases, and femoral cartilage lesions were found in 20%. The acetabular chondral lesions were correctly identified in 89-94% of cases. CONCLUSION: MRI provides a useful assessment of patients in whom a femoroacetabular impingement is clinically suspected. A high-resolution, nonarthrographic technique can provide preoperative information regarding the presence and anatomic site of labral and cartilage abnormalities. PMID- 17114530 TI - MRI of the sacroiliac joints in patients with moderate to severe ankylosing spondylitis. AB - OBJECTIVE: The objectives of our study were to evaluate whether MRI findings of the sacroiliac joints are able to distinguish between active and inactive disease in patients with established ankylosing spondylitis and to determine whether these findings correlate with markers of clinical activity, disease duration, severity, and degree of radiographic damage. MATERIALS AND METHODS: Eighteen patients with symptomatic moderate to severe ankylosing spondylitis were evaluated. MRI of the sacroiliac joint (1.5 T) was performed using fat-saturated T2-weighted, T1-weighted, STIR, and fat-saturated contrast-enhanced T1-weighted sequences. The sacroiliac joints were evaluated by two radiologists for enhancement, subchondral bone marrow edema, erosions, and subchondral fatty marrow infiltration. Findings on MRI were analyzed for correlation with multiple clinical characteristics and measures of disease activity, including radiographic scoring. RESULTS: In 17 patients, MRI showed abnormal findings of the sacroiliac joint. Ten patients showed active disease on MRI as measured by abnormal enhancement and subchondral bone marrow edema. Disease activity detected using MRI correlated in a positive fashion with only C-reactive protein (CRP) level. There was no correlation with the other measures of disease activity or with disease duration. In 14 patients, fatty subchondral bone marrow was detected on MRI. These changes were seen in patients with active and chronic disease and correlated with higher radiographic scores but not with disease duration or markers of disease activity. CONCLUSION: Contrast-enhanced MRI of the sacroiliac joint is sensitive in depicting sacroiliitis in patients with established ankylosing spondylitis. Subchondral edema and enhancement correlate with high CRP levels. Subchondral fatty bone marrow changes were seen in both active and chronic sacroiliitis and are correlated with higher radiographic scores; these changes may be a marker of more advanced disease. PMID- 17114531 TI - Therapeutic effect and outcome predictors of sciatica treated using transforaminal epidural steroid injection. AB - OBJECTIVE: The objectives of this retrospective study were to assess the therapeutic effect of transforaminal epidural steroid for sciatica and to identify outcome predictors. MATERIALS AND METHODS: Transforaminal epidural steroid injections were performed in 248 patients from June 2003 to May 2004. Fifty-six patients (33 women, 23 men; mean age, 53.3 years; age range, 30-83 years) were included. Therapeutic effects were evaluated 2 weeks after injection. The possible outcome predictors were as follows: intraepineural or extraepineural injection, saddle-type distribution pattern (contrast material distributed rostrally to the epidural portion of the preganglionic nerve root) or not saddle type, cause of sciatica (spinal stenosis vs herniated disk), patient age, patient sex, and duration of sciatica (acute or subacute [< 6 months] vs chronic [> 6 months]). The relationships between possible outcome predictors and therapeutic effects were analyzed. Statistical analysis was performed using Fisher's exact test, the chi-square test, and multiple logistic regression analysis. RESULTS: Forty-three (76.8%) of the 56 patients achieved a satisfactory result 2 weeks after transforaminal epidural steroid injection. Nineteen (65.5%) of the 29 patients treated by intraepineural injection and 24 (88.9%) of the 27 patients treated by extraepineural injection achieved a satisfactory result, and this difference was significantly different (p < 0.05). Other possible predictors of a better outcome were identified--that is, saddle-type pattern of contrast distribution, a herniated disk, and sciatica of less than 6 months' duration. Multiple regression analysis showed that the only factor significantly associated with outcome was the type of injection (p = 0.04, odds ratio: 5.01). CONCLUSION: Transforaminal epidural steroid is an effective tool for managing sciatica, and an extraepineural injection may be a predictor of a better outcome for sciatica treated using transforaminal epidural steroid. PMID- 17114532 TI - A novel approach to flexor hallucis longus tenography. AB - OBJECTIVE: This article presents a technically simple and more accurate approach to flexor hallucis longus (FHL) tenography than any we found reported in the literature. CONCLUSION: Tenography is used to evaluate and treat tenosynovitis. Standard FHL tenography protocol involves either direct percutaneous access of the FHL synovial sheath posterior to the medial malleolus or indirect filling of the FHL sheath from an injection of the flexor digitorum longus (FDL) tendon sheath, which often communicates with the FHL tendon sheath. However, with these methods, difficulty entering the FHL sheath may be encountered. We adapted our technique to access the FHL sheath as it courses below the sustentaculum talus. Our early experience with five cases using this technique reflects a 100% success rate with accurate needle placement within the FHL tendon sheath, thereby improving procedural efficiency. PMID- 17114533 TI - Using sonography for the early detection of elbow injuries among young baseball players. AB - OBJECTIVE: The aim of this study was to determine the usefulness of sonography for detecting elbow injuries among young baseball players. SUBJECTS AND METHODS: One hundred fifty-three volunteers ranging in age from 9 to 12 years and belonging to youth baseball teams participated. Sonography of the elbow was performed in the field when baseball exercises were being conducted. We analyzed the relationship between elbow pain and sonographic abnormalities and the relationship between pitchers and sonographic abnormalities. RESULTS: Sonography showed that 33 subjects had medial epicondylar fragmentation and two had early stage osteochondritis dissecans of the capitellum. In 25 subjects who agreed to further examination and treatment, radiography confirmed the sonographic findings. All of the 23 subjects with medial epicondylar fragmentation, who stopped throwing, obtained union of the bone and returned to baseball. The two subjects with osteochondritis dissecans of the capitellum underwent surgery before the osteochondral fragment became loosened. Sonographic abnormalities correlated with episodes of elbow pain. Pitchers statistically significantly had sonographic abnormalities. CONCLUSION: Sonography in the field can provide an opportunity to detect and treat elbow injuries before they become more advanced. PMID- 17114534 TI - Iterative decomposition of water and fat with echo asymmetry and least-squares estimation (IDEAL) fast spin-echo imaging of the ankle: initial clinical experience. AB - OBJECTIVE: Reliable, uniform fat suppression is important. Multiple approaches currently exist, many of which suffer from either suboptimal signal-to-noise ratio (SNR), or the inability to obtain consistent fat suppression around the ankle joint. Our purpose was to test iterative decomposition of water and fat with echo asymmetry and the least-squares estimation (IDEAL) method in combination with fast spin-echo imaging, which is able to achieve reliable high SNR images with uniform fat-water separation. SUBJECTS AND METHODS: We compared IDEAL fast spin-echo with conventional fat-suppressed fast spin-echo imaging in 33 ankles in 32 patients. Quantitative measurements of SNR and contrast-to-noise ratio efficiency were made, and qualitative diagnostic image quality and fat suppression scores were determined. RESULTS: We found that the SNR efficiency for both cartilage and fluid was similar for both techniques, and fluid/cartilage contrast-to-noise ratio efficiency was higher with IDEAL fast spin-echo imaging. Fat suppression and diagnostic quality scores using the IDEAL method were superior (p < 0.01) to fat-suppressed fast spin-echo imaging. CONCLUSION: IDEAL fast spin-echo imaging is a promising technique for MRI of the ankle. PMID- 17114535 TI - Sensitivity and specificity in detection of labral tears with 3.0-T MRI of the shoulder. AB - OBJECTIVE: MRI of the shoulder has been found to be sensitive and specific for detection of labral tears at 1.5 T or lower field strength compared with arthroscopy, whereas 3.0-T MRI of the shoulder has not been specifically assessed. This study assesses the sensitivity and specificity of MRI at 3.0 T for labral tears compared with arthroscopy. CONCLUSION: MRI of the shoulder at 3.0 T is very sensitive and specific compared with arthroscopy in detection of superior, anterior, and posterior labral tears. PMID- 17114536 TI - An international survey of hospital practice in the imaging of acute scaphoid trauma. AB - OBJECTIVE: Scaphoid fractures are relatively common. If not treated promptly there may be risk of long-term disability. However, unnecessary wrist immobilization is inconvenient and may hinder professional activities. Therefore, early accurate diagnosis is essential. Currently, the American College of Radiology deems MRI and radiographs as the most appropriate investigations in imaging acute scaphoid trauma. Our objective was to assess scaphoid imaging protocols. MATERIALS AND METHODS: To assess scaphoid imaging protocols, an international survey of imaging practice was performed. Two hundred hospitals worldwide were sent a survey regarding their scaphoid trauma imaging protocols. Only replies from hospitals that had full CT, MRI, and scintigraphy facilities were accepted. RESULTS: Data were obtained from 105 hospitals, of which 23 had fixed protocols. The number of scaphoid radiographic views varied from two to six. Before second-line investigations were initiated, repeat radiographs were usually performed in 76 of the 105 hospitals. In 29 hospitals, other imaging techniques were used without further radiography. The usual second-line investigation was MRI in 31/105, CT in 19/105, and scintigraphy in 14/105. Further protocols included CT or MRI in 10/105, CT or scintigraphy in 6/105, scintigraphy or MRI in 6/105, CT then MRI (if CT was negative) in 1/105, both CT and scintigraphy in 1/105, and scintigraphy then CT (if positive) in 1/105. There was equal preference among MRI, CT, and scintigraphy in 10/105 centers, and clinical examination and radiographs were used alone in 6/105. CONCLUSION: The survey reveals marked inconsistency in the imaging of acute scaphoid injury. Although other factors may have played a role, limited scientific evidence regarding the ideal imaging in acute scaphoid trauma may be the root of this inconsistency. PMID- 17114537 TI - Sonography and MRI of rectus abdominis muscle strain in elite tennis players. AB - OBJECTIVE: The objective of this study was to describe the imaging findings at sonography and MRI of rectus abdominis muscle strain in tennis players. CONCLUSION: Asymmetrical hypertrophy of the recti is seen in elite tennis players. The muscle belly hypertrophies on the side opposite the dominant arm and is subject to muscle tears of its deep fibers below the umbilicus. Imaging can be used to show these injuries. PMID- 17114538 TI - Segmental arterial mediolysis: CTA findings at presentation and follow-up. AB - OBJECTIVE: Segmental arterial mediolysis is a rare noninflammatory vascular disease of the abdominal splanchnic arteries. The purpose of our study was to retrospectively describe the CT angiography (CTA) findings of this disease and the evolution of those findings over time in five patients. CONCLUSION: Comparison of CTA and digital subtraction angiography suggests that CTA is useful to diagnose symptomatic segmental arterial mediolysis. Midterm CTA follow-up (median, 3 years) indicates that segmental arterial mediolysis lesions may resolve or remain unchanged. PMID- 17114539 TI - CAD in screening mammography. AB - OBJECTIVE: The benefit and cost of computer-assisted detection (CAD) mammography screening remains a topic of great interest in breast imaging. Our purpose is to reflect on and interleave two articles in this issue of the AJR that highlight the difficulty in assessing the actual benefit of using CAD from either retrospective or prospective studies. CONCLUSION: This commentary describes the possible benefit and some of the issues associated with the clinical use of current CAD technology while emphasizing the expectation of and need for future improvements in CAD performance. PMID- 17114540 TI - Testing the effect of computer-assisted detection on interpretive performance in screening mammography. AB - OBJECTIVE: The objective of our study was to test whether the use of computer assisted detection (CAD) improves sensitivity at no cost to specificity for the detection of breast cancer and enables more accurate assessment of fatty breast tissue compared with dense breast tissue. MATERIALS AND METHODS: We created a stratified random sample of screening mammograms weighted with difficult cases split evenly among women with fatty breast tissue and those with dense breast tissue: 114 patients were cancer-free, 114 had cancer 1 year after screening, and 113 had cancer 13-24 months after screening. In test settings 6 months apart, 19 community radiologists interpreted 341 bilateral screening mammograms with and without CAD. We compared the sensitivity and specificity using regression models adjusting for repeated measures. RESULTS: CAD assistance did not affect overall sensitivity (cancer by 1 year: 63.2% without CAD and 62.0% with CAD; cancer in 13 24 months: 33.5% without CAD and 32.3% with CAD), but its effect differed for visible masses that were marked by CAD compared with those that were not marked by CAD (hereafter referred to as "unmarked"). CAD was associated with improved sensitivity for marked visible cancers and decreased sensitivity for unmarked visible masses; the sensitivities without and with CAD, respectively, were as follows: marked cancer by 1 year, 82.7% versus 83.1%; marked cancer in 13-24 months, 44.2% versus 57.9%; unmarked cancer by 1 year, 37.4% versus 30.1%; unmarked cancer in 13-24 months, 29.7% versus 23.0% (p < 0.03 for both interactions between assistance and CAD marking for cancer by 1 year and cancer in 13-24 months). CAD marked 77% (70/91) of the visible cancers by 1 year and 67.3% (37/55) of the visible cancers in 13-24 months. CAD marked more visible calcified lesions (86%) than masses and asymmetric densities (67%) (p < 0.05). Overall specificity was 72% without and 75% with CAD (p < 0.02). CAD had a greater effect on both specificity (p < 0.02) and sensitivity (p < 0.03) among radiologists who interpret more than 50 mammograms per week. The results were the same for fatty breast tissue and dense breast tissue. CONCLUSION: In this experiment, CAD increased interpretive specificity but did not affect sensitivity because visible noncalcified lesions that went unmarked by CAD were less likely to be assessed as abnormal by radiologists. Breast density did not affect CAD's performance. PMID- 17114541 TI - Prospective assessment of computer-aided detection in interpretation of screening mammography. AB - OBJECTIVE: The purpose of this study was to prospectively assess the usefulness of computer-aided detection (CAD) in the interpretation of screening mammography and to provide the true sensitivity and specificity of this technique in a clinical setting. SUBJECTS AND METHODS: Over a 26-month period, 5,016 screening mammograms were interpreted without, and subsequently with, the assistance of the iCAD MammoReader detection system. Data collected for actionable findings included dominant feature (calcification, mass, asymmetry, architectural distortion), detection method (radiologist only, CAD only, or both radiologist and CAD), BI-RADS assessment code, associated histopathology for those undergoing biopsy, and tumor stage for malignant lesions. The study population was cross checked against an independent reference standard to identify false-negative cases. RESULTS: Of the 5,016 cases, the recall rate increased from 12% to 14% with the addition of CAD. Of the 107 (2%) patients who underwent biopsy, 101 (94%) were prompted by the radiologist and six (6%) were prompted by CAD. Of the 124 biopsies performed on actionable findings in the 107 patients, findings in 79 (64%) were benign and in 45 (36%) were in situ or invasive carcinoma. Three study participants who were not recalled by the radiologist with the assistance of CAD developed cancer within 1 year of the screening mammogram and were considered to be false-negative cases. The radiologist detected 43 (90%) of the 48 total malignancies and 45 (94%) of the 48 malignancies with the assistance of CAD. CAD missed eight cancers that were detected by the radiologist, which presented as architectural distortions (n = 3), irregular masses (n = 4), and a circumscribed mass (n = 1). CAD detected two in situ cancers as a faint cluster of calcifications that had not been perceived by the radiologist and one mass that was dismissed by the radiologist, accounting for at least a 4.7% increase in cancer detection rate. Sensitivity of screening mammography with the use of CAD (94%) represented an absolute and relative 4% increase over the sensitivity of the radiologist alone (90%). Specificity of screening mammography with and without the use of CAD was 99%. CONCLUSION: Routine use of CAD while interpreting screening mammograms significantly increases recall rates, has no significant effect on positive predictive value for biopsy, and can increase cancer detection rate by at least 4.7% and sensitivity by at least 4%. This study provides "true" values for sensitivity and specificity for use of CAD in interpretation of screening mammography as measured prospectively in the context of a working clinical setting. PMID- 17114542 TI - Full-field digital mammography on LCD versus CRT monitors. AB - OBJECTIVE: Our purpose was to determine if the display of full-field digital mammograms on a 5-megapixel liquid crystal display (LCD) monitor is at least equivalent to the display of the same on a 5-megapixel cathode ray tube (CRT) monitor. MATERIALS AND METHODS: Five radiologists evaluated normal anatomy and features of 61 abnormalities in 48 full-field digital mammograms. A 9-point Likert scale was used to compare images on two identical soft-copy review workstations, one equipped with two 5-megapixel CRTs and the other with two 5 megapixel LCDs. Outcomes were evaluated using a random-effects analysis of variance model. Means and SEs were reported. Ninety-five percent confidence intervals and p values were calculated. RESULTS: The two systems were equivalent for most features. The LCDs were rated better for the sharpness of mass margins (p = 0.011) and mass conspicuity (p = 0.050). For calcium features, the LCDs were rated better than the CRTs for lesion conspicuity (p = 0.010) and number of calcifications (p = 0.043). For architectural distortions, there was no statistically significant difference between the monitors in any of the features evaluated. For display characteristics, the LCDs were better for luminance (p = 0.021). The CRTs were significantly better for image noise (p = 0.001). In the overall ratings, there was no statistically significant difference between the two displays. CONCLUSION: The 5-megapixel monochrome active-matrix LCD is equivalent to and in some respects better than the 5-megapixel CRT display for full-field digital mammograms over a range of normal and abnormal findings. PMID- 17114543 TI - Comparison of MRI and sonography in the preliminary evaluation for fibroid embolization. AB - OBJECTIVE: The purpose of our study was to evaluate whether pelvic MRI provides additional clinically relevant information after sonography in the preprocedure evaluation of uterine artery embolization of fibroids. MATERIALS AND METHODS: Forty-nine women who presented for consultation for uterine artery embolization were retrospectively reviewed. The MRI and sonography scans were independently evaluated and compared for uterine size, fibroid size and location (categorized as paraendometrial, intramural, subserosal, or pedunculated) of the four largest fibroids in each patient, and the total number of fibroids present. RESULTS: One hundred twenty-two fibroids were measured. The uterine volume was significantly smaller as measured on MRI compared with sonography (p = 0.01). We found good MRI and sonography correlation of the volume of the single largest fibroid in each patient (R = 0.87) but poor correlation of fibroid location (R = 0.17). MRI detected 31 paraendometrial fibroids and three pedunculated fibroids that were thought to be intramural fibroids on sonography. Five fibroids thought to be paraendometrial on sonography were confirmed to be subserosal or intramural on MRI. Discrepancy in the total number of fibroids was noted, with additional fibroids found on MRI in 31 of 49 patients and erroneously suspected on sonography in five of 49 patients. Pelvic MRI affected management in 11 of 49 patients, leading to cancellation of uterine artery embolization in four patients. In another seven patients who were originally thought to be poor candidates on the basis of sonographic findings, uterine artery embolization was performed. MRI did not alter the management plan in 38 patients. CONCLUSION: MRI provided considerable additional information compared with sonography and affected clinical decision making in a substantial number of patients. MRI should be considered in all patients being evaluated for uterine artery embolization. PMID- 17114544 TI - Evaluation of real-time single-shot fast spin-echo MRI for visualization of the fetal midline corpus callosum and secondary palate. AB - OBJECTIVE: The objective of our study was to assess the visibility of the fetal corpus callosum and soft palate on standard single-shot fast spin-echo (SSFSE) imaging versus real-time (RT) SSFSE imaging. SUBJECTS AND METHODS: Part 1 of the study was a prospective analysis using a questionnaire rating the ease of use and utility of RT imaging. Part 2 of the study was a retrospective analysis of 69 fetal MRI studies with RT sagittal midline imaging of the head, face, or both. Standard and RT SSFSE image sets were de-identified, randomized, and shown to three pediatric neuroradiologists who rated on a 5-point scale whether the images were midline and how well they could see and characterize as normal the corpus callosum and secondary palate. The imaging results were correlated with postnatal diagnosis. Statistical methods included the Wilcoxon's signed rank test, McNemar chi-square test, and analysis of variance. RESULTS: Prospectively, the RT SSFSE technique was ranked as excellent in all the categories assessed. Retrospective analysis showed that the midline view obtained with RT SSFSE imaging was helpful in diagnosing the normal and abnormal secondary palate, allowing improved diagnosis of 19 (30.6%) of 62 cases of normal palate and four (57.1%) of seven cases of abnormal palate, when compared with the standard SSFSE technique. RT SSFSE imaging improved the ability to diagnose a normal corpus callosum on the midline view in 13 (27.6%) of 47 fetuses of 20 or more weeks gestational age. CONCLUSION: The RT SSFSE technique can aid in obtaining images in planes that are critical to the evaluation of a moving fetus, particularly when a midline sagittal view of the corpus callosum or palate is required. The use of this technique may lead to improved diagnosis of CNS or orofacial abnormalities in fetuses. PMID- 17114545 TI - MDCT of pancreatic adenocarcinoma: optimal imaging phases and multiplanar reformatted imaging. AB - OBJECTIVE: The objective of our study was to evaluate the individual contributions of arterial, pancreatic parenchymal, and portal venous phase (PVP) images and the utility of coronal and sagittal multiplanar reformatted (MPR) images in the assessment of pancreatic adenocarcinoma using triple-phase MDCT. MATERIALS AND METHODS: Thirty-one patients with and 35 patients without pancreatic adenocarcinoma underwent triple-phase MDCT. Three radiologists independently attempted to detect pancreatic adenocarcinoma and assess local extension using the MDCT images in five sessions. The first three sessions involved sets of images obtained in arterial phase, pancreatic parenchymal phase, and PVP separately and respectively. In the fourth session, a combination of axial images from all phases was evaluated. During the fifth session, radiologists had access to coronal and sagittal MPR images together with the axial images obtained in all phases. Results were compared with surgical findings using receiver operating characteristic (ROC) analysis and kappa statistics. RESULTS: Regarding tumor detection, the image set composed of coronal and sagittal MPR images and of axial images obtained in all phases had a significantly higher value for the area under the ROC curve (A(Z), 0.98 +/- 0.01) than the other image sets and yielded the highest sensitivity (93.5%). The sensitivity of the arterial phase image set (80.6%) was significantly lower than that of all other image sets. Whereas the image set composed of coronal and sagittal MPR images and axial images obtained in all phases yielded the highest kappa values for all local extension factors evaluated, the image set composed of only arterial phase images yielded the lowest kappa values for almost all of the factors. CONCLUSION: A combination of pancreatic parenchymal phase and PVP imaging is necessary and efficient for the assessment of pancreatic adenocarcinoma. The addition of coronal and sagittal MPR images increased the performance of MDCT, especially in the evaluation of local extension. PMID- 17114546 TI - ADC measurement of abdominal organs and lesions using parallel imaging technique. AB - OBJECTIVE: The purpose of our study was to assess the reliability and usefulness of parallel imaging for apparent diffusion coefficient (ADC) measurement of abdominal organs and lesions. MATERIALS AND METHODS: Single-shot spin-echo echo planar diffusion-weighted MRI (TE = 66, b = 0, 600 s/mm2) was performed in phantom and clinical studies. The b value was set to minimize the effects of perfusion in tissue and to maintain signal-to-noise ratio. Bottle phantoms were scanned with and without parallel imaging and with various parallel imaging factors and at various positions to evaluate the effects of parallel imaging on ADCs. In 200 consecutive clinical patients (122 men and 78 women: mean age, 61.9 years), ADCs were calculated for liver (four segments), spleen, pancreas (head, body, tail), gallbladder, renal parenchyma, and back muscle, and then compared to evaluate the reliability of clinical ADC measurements with parallel imaging. ADCs were also calculated for diffuse diseases and focal lesions (94 malignant and 93 benign) of abdominal organs to evaluate the clinical usefulness of ADC. RESULTS: Location-dependent changes in water ADCs were minimal with parallel imaging factors first of 3, then of 4, and were small except for measurements at the image periphery. Acetone ADCs were saturated at 4.00 x 10(-3) mm2/s. Degraded image quality prevented ADC measurement of the left hepatic lobe and pancreas in 7-18 patients. There was no significant difference among ADCs of four liver segments (1.50 +/- 0.24 [SD] x 10(-3) mm2/s - 1.56 +/- 0.31 x 10(-3) mm2/s) and between ADCs of the right and left kidneys (2.65 +/- 0.30 x 10(-3) mm2/s, 2.59 +/ 0.33 x 10(-3) mm2/s). ADC of the pancreas tail (1.65 +/- 0.37 x 10(-3) mm2/s) was significantly lower than those of the head (1.81 +/- 0.40 x 10(-3) mm2/s) and body (1.81 +/- 0.41 x 10(-3) mm2/s) (p < 0.005). Renal ADCs were significantly lower in patients with renal failure (right: 2.15 +/- 0.30 x 10(-3) mm2/s; left: 2.11 +/- 0.25 x 10(-3) mm2/s) than in those without disease (right: 2.67 +/- 0.29 x 10(-3) mm2/s; left: 2.60 +/- 0.32 x 10(-3) mm2/s) (p < 0.005). ADC of pancreatic cancer was significantly higher than that of healthy pancreas (p < 0.05). ADC of renal angiomyolipoma was significantly lower than those of renal cell carcinoma and healthy renal parenchyma (p < 0.0005). CONCLUSION: Clinical ADC measurements of abdominal organs and lesions using parallel imaging appear to be reliable and useful, and the effect of parallel imaging on calculated values is considered to be minimal. PMID- 17114547 TI - Fine-needle aspiration biopsy of solid pancreatic masses: comparison of CT and endoscopic sonography guidance. AB - OBJECTIVE: Imaging-guided biopsies of solid pancreatic masses are performed with either CT or endoscopic sonography at our institution. We compared test characteristics of fine-needle aspiration biopsies guided using CT with those guided using endoscopic sonography and secondarily evaluated for an effect of mass size. MATERIALS AND METHODS: Of 70 solid pancreatic masses, 43 (mean size, 4.4 cm; range, 1.5-10.3 cm) underwent fine-needle (20- to 22-gauge) aspiration biopsy with CT guidance and 27 (mean size, 2.3 cm; range, 1.0-5.0 cm) underwent fine-needle (22-gauge) aspiration biopsy with endoscopic sonography guidance. The diagnostic rate, sensitivity, and negative predictive value (NPV) for each technique were compared using Fisher's exact test before and after stratifying masses by size as small (< or = 3 cm) or large (> 3 cm). RESULTS: The overall diagnostic rate, sensitivity, and NPV of fine-needle aspiration biopsies guided using CT (97.7%, 94.9%, and 60%, respectively) were not significantly different from those guided using endoscopic sonography (88.9%, 85%, and 57.1%, respectively). Among small masses, the diagnostic rate and sensitivity for biopsies guided using CT (100% and 100%, respectively) were not significantly different from those for biopsies guided using endoscopic sonography (90.9% and 93.8%, respectively). Among large masses, the diagnostic rate and sensitivity (96.6% and 92.3%, respectively) for biopsies guided using CT were not significantly different from those for biopsies guided using endoscopic sonography (83.3% and 50%, respectively). CONCLUSION: When biopsying solid pancreatic masses with fine needles, procedures guided with CT and those guided with endoscopic sonography have similar test characteristics regardless of mass size. PMID- 17114548 TI - Congenital anomalies and normal variants of the pancreaticobiliary tract and the pancreas in adults: part 1, Biliary tract. AB - OBJECTIVE: The purpose of this article is to highlight the imaging features of congenital anomalies and normal variants of the biliary tract with contemporary imaging techniques such as MR cholangiopancreatography (MRCP), MRI, and helical CT. CONCLUSION: Recognizing findings of congenital anomalies and normal variants of the biliary tract at MRCP, MRI, and helical CT, and knowledge of the clinical significance of each entity, are important for establishing a correct diagnosis and in guiding appropriate clinical management. PMID- 17114549 TI - Congenital anomalies and normal variants of the pancreaticobiliary tract and the pancreas in adults: part 2, Pancreatic duct and pancreas. AB - OBJECTIVE: The purpose of this article is to highlight the imaging features of congenital anomalies and normal variants of the pancreatic duct and the pancreas using contemporary imaging techniques such as MR cholangiopancreatography (MRCP), MRI, and helical CT. CONCLUSION: Congenital anomalies and normal variants of the pancreatic duct and the pancreas may be clinically significant and may create a diagnostic challenge. Recognition of the updated imaging features of these entities is important in clinical management and for avoiding misdiagnosis. PMID- 17114550 TI - Preoperative staging of rectal cancer: comparison of 3-T high-field MRI and endorectal sonography. AB - OBJECTIVE: The aim of this study was to compare phased-array 3-T MRI and endorectal sonography in the preoperative staging of rectal cancer. MATERIALS AND METHODS: During an 8-month period, 24 patients with rectal cancer underwent both 3-T MRI performed with phased-array coils and 7.5- to 10-MHz endorectal sonography in the 3 weeks before surgical resection. Three radiologists independently reviewed the MR and endorectal sonographic images. The histopathologic findings in resected specimens were used to evaluate the sensitivities and specificities of these techniques for invasion of the muscularis propria and perirectal tissue and for lymph node involvement. Receiver operating characteristic (ROC) analysis was used to compare the diagnostic accuracies of the techniques. RESULTS: For muscularis propria invasion, the mean sensitivities of both MRI and endorectal sonography were 100%, and the mean specificities were 66.7% and 61.1%, respectively. The differences in the mean sensitivities and specificities were not statistically significant (p > 0.05 in each case). For perirectal tissue invasion, MRI and endorectal sonography had comparable sensitivities and specificities (91.1% vs 100%, 92.6% vs 81.5%; p > 0.05 in each case). They also had similar sensitivities and specificities for lymph node involvement (63.6% vs 57.6%, 92.3% vs 82.1%; p > 0.05 in each case). ROC curves for muscularis propria invasion and lymph node involvement showed no differences in diagnostic accuracy. The mean area under the ROC curve for endorectal sonography (A(Z) = 0.996) for perirectal tissue invasion, however, showed higher accuracy than that of MRI (A(Z) = 0.938, p = 0.028). CONCLUSION: The sensitivity, specificity, and accuracy of 3-T MRI were similar to those of endorectal sonography for muscularis propria invasion and lymph node involvement, but for perirectal tissue invasion, 3-T MRI was less accurate than endorectal sonography. PMID- 17114551 TI - Isotropic 3D T2-weighted MR cholangiopancreatography with parallel imaging: feasibility study. AB - OBJECTIVE: The aim of this study was to compare the quality of images obtained with fast 3D T2-weighted turbo spin-echo (TSE) MR cholangiopancreatography (MRCP) sequences and 1-mm isotropic voxels with the quality of conventional 2D MRCP images. SUBJECTS AND METHODS: Thirty consecutively registered patients (14 women, 16 men; average age, 60.2 years; age range, 32-87 years) underwent imaging at 1.5 T with a 6-element body array coil. All imaging was performed with three MRCP techniques: free-breathing 3D T2-weighted TSE (TR/TE, 1,300/680; flip angle, 180 degrees; field of view, 250-300 mm; matrix size, 256 x 256; slice thickness, 1 mm; parallel acquisition technique factor, 2); breath-hold 3D T2-weighted TSE (same parameters as the free-breathing 3D technique); breath-hold coronal and oblique coronal thick-slab 2D TSE without parallel acquisition technique (2,800/1,100; flip angle, 150-180 degrees). Quantitative measures of image signal and contrast were evaluated by analysis of variance and paired Student's t tests. A 5-point scale (1, nondiagnostic, to 5, high diagnostic confidence) was used to compare the 3D and 2D data sets for image quality and definition of biliary and pancreatic ductal anatomic features. Friedman's nonparametric and Wilcoxon's rank sum tests were performed for statistical analysis of the qualitative assessments. RESULTS: Quantitative results showed free-breathing and breath-hold 3D TSE images had significantly higher relative signal intensity and contrast than 2D TSE images (p < 0.0001). The qualitative findings showed that both free-breathing and breath-hold 3D TSE techniques gave better delineation of biliary anatomy (p < 0.0001) than the 2D technique. The overall quality of 3D images was better than that of 2D images, and 3D imaging was better at depicting pancreatic ducts, although the difference did not reach statistical significance. CONCLUSION: Three dimensional volumetric MRCP images are of superior quality and give better delineation of pancreaticobiliary anatomy than conventional 2D images and have the added advantage of multiplanar and postprocessing capabilities. PMID- 17114552 TI - CT findings of cholangiocarcinoma associated with recurrent pyogenic cholangitis. AB - OBJECTIVE: The purpose of our study was to determine the characteristic CT findings of cholangiocarcinoma associated with recurrent pyogenic cholangitis. CONCLUSION: CT findings of cholangiocarcinoma associated with recurrent pyogenic cholangitis are important in order to improve early diagnosis and proper treatment. Cholangiocarcinoma associated with recurrent pyogenic cholangitis is predominantly located in the atrophic hepatic lobes and in the hepatic lobes of biliary calculi and is associated with the narrowing or obliteration of the portal vein. PMID- 17114553 TI - Radiologic removal and replacement of port-catheter systems for hepatic arterial infusion chemotherapy. AB - OBJECTIVE: The purpose of our study was to retrospectively evaluate the safety and efficacy of radiologic removal and replacement of port-catheter systems. MATERIALS AND METHODS: Between January 1999 and December 2004, 532 patients with unresectable advanced liver cancer underwent radiologic placement of port catheter systems at our institution. Of these, 18 patients (nine men and nine women; age range, 32-83 years; mean age, 53.8 years) underwent removal of an implanted port-catheter system via the right femoral artery and radiographically guided replacement with a new system to allow continuous hepatic arterial infusion chemotherapy; we retrospectively reviewed these 18 cases. The reasons for removal of the previously implanted systems were as follows: catheter dislodgement (n = 15), catheter obstruction (n = 1), infection related to the implanted port (n = 1), and hemodynamic change (n = 1). Digital subtraction angiography and CT were performed, usually during injection of contrast medium through the implanted port-catheter system, within a few days after the replacement procedure and every 3 months thereafter. RESULTS: We successfully performed radiologic removal and replacement of the portcatheter system while the patient was under local anesthesia in all 18 patients without complications requiring treatment. The cumulative patency rates of the hepatic artery after removal of the old port-catheter system and replacement with a new port-catheter system were 87.8% and 64.1% at 6 months and 1 year, respectively. Hepatic arterial infusion chemotherapy after replacement was performed 0-68 times (median, 19 times). CONCLUSION: When an implanted port-catheter system can no longer be used but the patency of the hepatic artery is confirmed and continuous hepatic arterial infusion chemotherapy is required, removal and replacement of the port-catheter system are recommended. PMID- 17114554 TI - Percutaneous needle aspiration of multiple pyogenic abscesses of the liver: 13 year single-center experience. AB - OBJECTIVE: The aim of this study was to report 13 years of experience in the management of multiple pyogenic liver abscesses with only percutaneous needle aspiration of the abscess cavities under sonographic guidance. MATERIALS AND METHODS: From December 1991 to October 2004, 39 consecutively registered patients (29 men and 10 women; age range, 37-82 years; mean age, 64 years) with 118 pyogenic liver abscesses were treated with sonographically guided percutaneous needle aspiration at our institution. The number of pyogenic liver abscesses per patient ranged from two to 15 (mean, 3.0). RESULTS: Eighty-seven percutaneous needle aspirations were performed on 39 patients with 118 pyogenic liver abscesses (range, 1-4 aspirations per patient; mean, 2.2 aspirations per patient). Because they were close to another aspirated abscess in the right lobe of the liver, 31 (26.3%) of 118 abscesses were aspirated without removal of the needle from the liver. Thirty-six (92.3%) of 39 patients were treated with a single aspiration of an abscess in a single session. The other three patients needed two aspiration sessions. No patient needed imaging-guided percutaneous catheter drainage or open surgical drainage. Complete reconstitution of liver parenchyma occurred within a maximum of 80 days. No abscesses recurred during the follow-up period, which ranged from 7 to 42 months (mean, 18 months). CONCLUSION: Percutaneous needle aspiration of multiple pyogenic abscesses under sonographic guidance is a safe, effective, and low-cost procedure. In our experience, percutaneous needle aspiration was acceptable to patients. Our data suggest that a trial of percutaneous needle aspiration should always be undertaken before catheter drainage or surgery. PMID- 17114555 TI - Imaging-guided catheter drainage of abdominal collections with fistulous pancreaticobiliary communication. AB - OBJECTIVE: Our aim was to study the success of percutaneous imaging-guided catheter drainage of abdominal collections with documented fistulous pancreaticobiliary communication. MATERIALS AND METHODS: Fifty-seven patients (age range, 23-88 years) with abdominal collections who underwent imaging-guided catheter drainage were included in this retrospective study. These collections showed communication with either the pancreatic duct (n = 15) or the biliary duct (n = 42) on imaging. The imaging guidance included CT (n = 40), sonography (n = 17), and fluoroscopy (n = 4), either alone or in combination. The success of catheter drainage was described as resolution of the collection on follow-up imaging and clinical improvement. Other treatments directed toward management of leaks or collections were also recorded. RESULTS: The success rates of catheter drainage for abdominal collections with biliary and pancreatic ductal communication were 93% (39/42) and 67% (10/15), respectively. The difference was statistically significant (p = 0.01). The three complications seen in this study were pneumothorax in one patient, bowel perforation in one, and death in one. The drainage catheter was upsized in five patients and an additional catheter was needed in nine patients. CONCLUSION: Imaging-guided catheter drainage is a clinically useful option for drainage of abdominal collections that have pancreaticobiliary communications. The success rate is significantly better for collections with biliary communication than for those with pancreatic communication. PMID- 17114556 TI - MDCT of right ventricular function: comparison of right ventricular ejection fraction estimation and equilibrium radionuclide ventriculography, part 1. AB - OBJECTIVE: The aim of this study was to calculate right ventricular ejection fraction by use of ECG-gated MDCT and to compare the results with those of equilibrium radionuclide ventriculography. SUBJECTS AND METHODS: Forty-nine consecutively examined patients (30 men, 19 women; mean age, 59 years) with known or suspected right ventricular dysfunction secondary to bronchopulmonary (n = 30) or pulmonary vascular (n = 19) disease underwent ECG-gated 16-MDCT angiography of the heart (rotation time, 0.42 second; 120 kV; 300 mAs; collimation, 12 x 0.75 mm; pitch, 0.2) after CT angiographic examination of the entire thorax according to a standard protocol. Biphasic administration of a 30% contrast agent was systematically performed (phase 1, 90 mL at 3 mL/s; phase 2, 30 mL at 1.5 mL/s); no patient received additional medication. Right ventricular ejection fraction was calculated after two reviewers in consensus determined the reconstruction windows and segmentation of the right ventricular cavity on a series of diastolic and systolic short-axis images. The results were compared with those of equilibrium radionuclide ventriculography. RESULTS: At data acquisition, the mean (+/- SD) heart rate of the study group was 82 +/- 13.87 beats per minute (BPM) (range, 51-115 BPM). ECG showed a sinus rhythm in 30 (61%) of the patients and irregular cardiac rhythm in 19 (39%) of the patients. Agreement between the two techniques was estimated by intraclass correlation coefficient (0.77), the method of Bland and Altman (limits of concordance, -14.9 and 13.7), and percentage of variability between two measurements expressed by mean absolute percentage error (12.1%). The estimated effective dose for heart examination was 7.48 mSv with CT and 5 mSv with scintigraphy. The mean effective dose for the chest and heart CT examinations was 11.64 mSv. CONCLUSION: Right ventricular ejection fraction can be reliably estimated with 16-MDCT in unselected patients. PMID- 17114557 TI - MDCT of right ventricular function: impact of methodologic approach in estimation of right ventricular ejection fraction, part 2. AB - OBJECTIVE: The purpose of this study was to evaluate the impact of the methodologic approach for MDCT estimation of right ventricular ejection fraction (RVEF). MATERIALS AND METHODS: In 49 consecutive patients (30 men, 19 women; mean age, 59 years) known to have or suspected of having right ventricular (RV) dysfunction secondary to pulmonary disease, 16-MDCT of the heart was performed after standard CT angiographic examination of the entire thorax, with determination of RVEF by two reviewers who had limited experience in cardiac CT. The reconstruction windows were determined using the ECG tracing (reviewer 1) or using transverse test images obtained in 5% steps through the entire R-R interval showing the largest and smallest RV cavity areas (reviewer 2). After manual segmentation of the ventricular cavity on diastolic and systolic short-axis reformations by each reviewer, the end-diastolic and end-systolic RV volumes were calculated, with subsequent determination of the RVEF. CT results were compared with those of equilibrium radionuclide ventriculography. RESULTS: Agreement between the two methods for determining the end-systolic and end-diastolic phases was observed in 61% of cases (n = 30) for the systole and 59% of cases (n = 29) for the diastole. Discordant selections were observed in 39% of cases (n = 19) for determination of the systole and in 41% of cases (n = 20) for determination of the diastole, ranging from 5% to 15% of the R-R interval, suggesting that selection of the reconstruction window on the ECG tracing does not differ significantly from that obtained by the visual analysis of transverse test images. Focusing on the 59 common selections of the reconstruction windows made by the two reviewers, no statistically significant differences were found in the determination of mean (+/- SD) end-diastolic volumes (reviewer 1, 176.21 +/- 67 mL vs reviewer 2, 175.55 +/- 71.24 mL; p = 0.98) and end-systolic (reviewer 1, 97.3 +/- 26.49 mL vs reviewer 2, 96.33 +/- 65.72 mL; p = 0.65), suggesting the lack of operator dependence in the manual-contour drawing process. No significant difference was found between the mean values of RVEF obtained by each reviewer with MDCT and equilibrium radionuclide ventriculography, and there was excellent interobserver agreement with MDCT (intraclass correlation coefficient, 0.86). Using a Bland-Altman approach, the limits of concordance between the two reviewers ranged between -10.2 and 10.9. The mean absolute percentage error for measuring RVEF between the two reviewers was 9.7%. A moderate agreement was found between RVEFs obtained on CT by each reviewer and scintigraphy (intraclass correlation coefficients, 0.76 for reviewer 1 and 0.64 for reviewer 2). CONCLUSION: These results show that RVEF can be accurately assessed with ECG gated MDCT using commercially available software. PMID- 17114558 TI - 99mTc sestamibi uptake by acute pulmonary embolism. PMID- 17114559 TI - Normal CT appearance of the distal thoracic duct. AB - OBJECTIVE: The aims of this study were to determine how often the distal thoracic duct can be identified on neck CT and to characterize the CT appearance of the duct. MATERIALS AND METHODS: In a review of electronic medical records from January 2001 to January 2003 we identified the cases of 500 patients who had undergone CT of the neck. Because they had confounding factors such as cancer or cervical lymphadenopathy, 199 of these patients were excluded, leaving 301 patients in the study: 131 (44%) male patients and 170 (56%) female patients. The age range was 11-92 years (average age, 46 years). Two head and neck radiologists used strict diagnostic criteria and consensus to identify the distal thoracic duct on both sides of the neck. One half of the images selected at random were flipped left to right. The purpose of randomization was to avoid interpretation bias, because the thoracic duct is known to typically course within the left side of the neck. The configuration of the distal duct was tabulated, and effects of age and sex were statistically evaluated. RESULTS: The left side of the neck was unevaluable in 26 (9%) of 301 patients because of streak artifact. In the other 275 patients, the distal thoracic duct was identified in the left side of the necks of 150 (55%) of the patients. Eleven of these patients (4%) also had a visible duct in the right side of the neck, but a right-sided duct was never identified without a left-sided counterpart. The distal thoracic duct had a tubular configuration in 70 (43%), a flared configuration in 72 (45%), and a long segmental fusiform dilation in 19 (12%) of 161 patients. Patient sex had no significant effect on the appearance of the distal thoracic duct. Older patient age had a slight positive effect on the size of the duct. CONCLUSION: Familiarity with the normal CT appearance of the distal thoracic duct can be helpful in differentiating a normal duct from pathologic lesions of the lower neck, such as lymphadenopathy. PMID- 17114560 TI - Conscious sedation reduces distress in children undergoing voiding cystourethrography and does not interfere with the diagnosis of vesicoureteric reflux: a randomized controlled study. AB - OBJECTIVE: Voiding cystourethrography (VCU) is a distressing procedure for children. Conscious sedation using oral midazolam may reduce this distress, but its use may also alter the ability of the VCU to show vesicoureteric reflux (VUR). The objectives of our study were to assess the effectiveness of conscious sedation using oral midazolam when administered routinely in children undergoing VCU and to ensure that conscious sedation using oral midazolam does not alter the ability of VCU to show VUR. SUBJECTS AND METHODS: Our study was a randomized double-blind controlled trial performed at a university teaching hospital; our study group consisted of children over the age of 1 year who been referred for their first VCU examination from July 2001 to July 2003. Participants were randomized to receive a placebo or midazolam syrup (0.5 mg/kg) before the examination. The primary outcome measures were the Groningen Distress Rating Scale (GDRS) and grading of VUR, as defined by the international grading system established by the International Reflux Study Group. RESULTS: There were no serious adverse events. One hundred thirty-nine children were randomized in the study, and 117 underwent complete assessment. Eight who underwent VCU after the study day were included in a "complete case" intention-to-treat analysis. In the placebo group, 34 children (61%) experienced serious distress or severe distress (GDRS score, 3 or 4). In the midazolam group, 16 children (26%) experienced the same degree of distress. There was a significant difference between the GDRS scores (nonlinear mixed-model analysis, p < 0.001) of the two study groups. The number needed to treat to reduce serious or severe distress in one child was 2.9 (95% CI, 1.9-5.5). VUR was identified in 16% of all children. There was no difference in VUR grading between the groups (nonlinear mixed-model analysis, p = 0.31). CONCLUSION: Routine use of oral midazolam (0.5 mg/kg) for conscious sedation of children undergoing VCU reduces distress and does not alter the ability of VCU to show VUR well enough to allow diagnosis. PMID- 17114561 TI - Sonographic appearance of the epididymis in pediatric testicular torsion. AB - OBJECTIVE: The purpose of this study was to determine the frequency of an enlarged epididymis in pediatric patients with testicular torsion and to determine whether an altered epididymis might be a helpful ancillary sonographic sign of testicular torsion. MATERIALS AND METHODS: The sonograms of 50 pediatric patients (age range, neonate-17 years) with testicular torsion were retrospectively reviewed for the size, appearance, and blood flow of the epididymis. Medical records were reviewed for surgical and pathologic findings and to determine whether testicular salvage had been possible. RESULTS: The epididymis was enlarged (maximum dimension, 5.5 cm) in 47 of the 50 patients with acute or late phase torsion and after manual or spontaneous detorsion. The average difference in volume between the ipsilateral epididymis and the contralateral epididymis was 30 cm3, highly significant (p < 0.0001). The shape of the epididymis was altered in 92% of the cases (globular, bilobular, or multilobular). Seventy-three percent showed increased echogenicity and 27% appeared isoechoic. Of those with active torsion, 93% of the epididymides were avascular; 2%, hypovascular; and 5%, hypervascular. After detorsion, 100% of the epididymides (10/10) had blood flow. In patients with testicular loss due to infarction, pathology showed engorgement and enlargement of the epididymis with hemorrhagic infarction. In one patient with a hypervascular epididymis, surgery showed inflammation and erythema of the epididymis. Testicular loss occurred in each type of epididymal flow pattern. CONCLUSION: A markedly enlarged, echogenic, and avascular or hypovascular epididymis is an ancillary sonographic sign in pediatric patients with testicular torsion. A hypervascular enlarged epididymis infrequently occurs (5% of cases) and should not be mistaken for epididymitis. In addition, the return of epididymal blood flow is an ancillary sign of successful testicular detorsion. PMID- 17114562 TI - Comparison of CT venography with MR venography in cerebral sinovenous thrombosis. AB - OBJECTIVE: The purpose of this study was to compare cerebral CT venography with MR venography and determine the reliability of CT venography in the diagnosis of cerebral sinovenous thrombosis. SUBJECTS AND METHODS: Fifty patients who were clinically suspected of having cerebral sinovenous thrombosis, irrespective of age and sex, underwent cerebral CT venography and MR venography. Projection venograms were displayed using maximum-intensity-projection images for both CT venography and MR venography. The CT venograms were also displayed using the integral algorithm, which depicts the average intensity value of the first five voxels deep in relation to the model surface that is nearest the viewer, allowing direct visualization of the thrombus in the sinuses. All CT venograms and MR venograms were independently evaluated by experienced neuroradiologists. RESULTS: Of these 50 patients, 30 patients were diagnosed as having cerebral sinovenous thrombosis on both CT venography and MR venography. The total numbers of sinuses involved were 81 and 77 (CT venography and MR venography). When MR venography was used as the gold standard, CT venography was found to have both a sensitivity and a specificity of 75-100%, depending on the sinus and vein involved. CONCLUSION: CT venography is as accurate as MR venography for diagnosing cerebral sinovenous thrombosis. PMID- 17114564 TI - CT and MRI of coronary artery disease: evidence-based review. AB - OBJECTIVE: The educational objective of this evidence-based self-assessment module is to use case examples to review the current evidence and the roles of CT and MRI in evaluating and managing patients with both congenital and acquired coronary artery disease. CONCLUSION: In this educational module, we review the use of CT and MRI in the noninvasive diagnosis and management of patients with coronary artery disease. PMID- 17114565 TI - AJR teaching file: Right ventricular mass presenting with a heart murmur. PMID- 17114566 TI - AJR teaching file: High-output cardiac failure in a patient with a history of hereditary hemorrhagic telangiectasia. PMID- 17114567 TI - Cocaine esterase: interactions with cocaine and immune responses in mice. AB - Cocaine esterase (CocE) is the most efficient protein catalyst for the hydrolysis of cocaine characterized to date. The aim of this study was to investigate the in vivo potency of CocE in blocking cocaine-induced toxicity in the mouse and to assess CocE's potential immunogenicity. Cocaine toxicity was quantified by measuring the occurrence of convulsions and lethality. Intravenous administration of CocE (0.1-1 mg) 1 min before cocaine administration produced dose-dependent rightward shifts of the dose-response curve for cocaine toxicity. More important, i.v. CocE (0.1-1 mg), given 1 min after the occurrence of cocaine-induced convulsions, shortened the recovery time after the convulsions and saved the mice from subsequent death. Effects of repeated exposures to CocE were evaluated by measuring anti-CocE antibody titers and the protective effects of i.v. CocE (0.32 mg) against toxicity elicited by i.p. cocaine (320 mg/kg) (i.e., 0-17% occurrence of convulsions and lethality). CocE retained its potency against cocaine toxicity in mice after a single prior CocE exposure (0.1-1 mg), and these mice did not show an immune response. CocE retained similar effectiveness in mice after three prior CocE exposures (0.1-1 mg/week for 3 weeks), although these mice displayed 10-fold higher antibody titers. CocE partially lost effectiveness (i.e., 33-50% occurrence of convulsions and lethality) in mice with four prior exposures to CocE (0.1-1 mg/2 week for four times), and these mice displayed approximately 100 fold higher antibody titers. These results suggest that CocE produces robust protection and reversal of cocaine toxicity, indicating CocE's therapeutic potential for acute cocaine toxicity. Repeated CocE exposures may increase its immunogenicity and partially reduce its protective ability. PMID- 17114568 TI - Presynaptic delta opioid receptors regulate ethanol actions in central amygdala. AB - Endogenous opioid systems are implicated in the reinforcing effects of ethanol consumption. For example, delta opioid receptor (DOR) knockout (KO) mice show greater ethanol consumption than wild-type (WT) mice (Roberts et al., 2001). To explore the neurobiological correlates underlying these behaviors, we examined effects of acute ethanol application in brain slices from DOR KO mice using whole cell patch recording techniques. We examined the central nucleus of amygdala (CeA) because the CeA is implicated in alcohol reinforcement (Koob et al., 1998). We found that the acute ethanol effects on GABA(A) receptor-mediated inhibitory postsynaptic currents (IPSCs) were greater in DOR KO mice than in WT mice. Ethanol increased the frequency of miniature IPSCs (mIPSCs) significantly more in DOR KO mice than in WT mice. In CeA of WT mice, application of ICI 174864 [[allyl]2-Tyr-alpha-amino-isobutyric acid (Aib)-Aib-Phe-Leu-OH], a DOR inverse agonist, augmented ethanol actions on mIPSC frequency comparable with ethanol effects seen in DOR KO mice. Superfusion of the selective DOR agonist D-Pen(2),D Pen(5)-enkephalin decreased the mean frequency of mIPSCs; this effect was reversed by the DOR antagonist naltrindole. These findings suggest that endogenous opioids may reduce ethanol actions on IPSCs of CeA neurons in WT mice through DOR-mediated inhibition of GABA release and that the increased ethanol effect on IPSCs in CeA of DOR KO mice could be, at least in part, due to absence of DOR-mediated inhibition of GABA release. This result supports the hypothesis that endogenous opioid peptides modulate the ethanol-induced augmentation of GABA(A) receptor-dependent circuitry in CeA (Roberto et al., 2003). PMID- 17114569 TI - Chromosomal abnormalities in marrow stromal cells from myelodysplastic syndromes (MDS). PMID- 17114570 TI - Discrete stem cells: subsets or a continuum? PMID- 17114571 TI - Increased mortality with FLA compared with ADE chemotherapy in high-risk AML. PMID- 17114572 TI - Osteonecrosis of the jaws in newly diagnosed multiple myeloma patients treated with zoledronic acid and thalidomide-dexamethasone. PMID- 17114573 TI - Use of the International System for Human Cytogenetic Nomenclature (ISCN). PMID- 17114574 TI - PASD1 is a potential multiple myeloma-associated antigen. PMID- 17114575 TI - A PHD finger protein involved in both the vernalization and photoperiod pathways in Arabidopsis. AB - The proper timing of flowering is critical for successful reproduction. The perception of the seasonal cues of day-length changes and exposure to cold influences flowering time in many plant species through the photoperiod and vernalization pathways, respectively. Here we show that a plant homeodomain (PHD) finger-containing protein, VIN3-LIKE 1 (VIL1), participates in both the photoperiod and vernalization pathways in Arabidopsis thaliana by regulating expression of the related floral repressors FLOWERING LOCUS C (FLC) and FLOWERING LOCUS M (FLM). In the vernalization pathway, VIL1, along with VERNALIZATION INSENSITIVE 3 (VIN3), is necessary for the modifications to FLC and FLM chromatin that are associated with an epigenetically silenced state and with acquisition of competence to flower. In addition, VIL1 regulates FLM independently of VIN3 in a photoperiod-dependent manner. PMID- 17114576 TI - Wnt/beta-catenin signaling regulates vertebrate limb regeneration. AB - The cellular and molecular bases allowing tissue regeneration are not well understood. By performing gain- and loss-of-function experiments of specific members of the Wnt pathway during appendage regeneration, we demonstrate that this pathway is not only necessary for regeneration to occur, but it is also able to promote regeneration in axolotl, Xenopus, and zebrafish. Furthermore, we show that changes in the spatiotemporal distribution of beta-catenin in the developing chick embryo elicit apical ectodermal ridge and limb regeneration in an organism previously thought not to regenerate. Our studies may provide valuable insights toward a better understanding of adult tissue regeneration. PMID- 17114578 TI - The molecular pathogenesis of pancreatic cancer: clarifying a complex circuitry. PMID- 17114577 TI - Reduced growth of Drosophila neurofibromatosis 1 mutants reflects a non-cell autonomous requirement for GTPase-Activating Protein activity in larval neurons. AB - Neurofibromatosis type 1 (NF1) is among the most common genetic disorders of humans and is caused by loss of neurofibromin, a large and highly conserved protein whose only known function is to serve as a GTPase-Activating Protein (GAP) for Ras. However, most Drosophila NF1 mutant phenotypes, including an overall growth deficiency, are not readily modified by manipulating Ras signaling strength, but are rescued by increasing signaling through the cAMP-dependent protein kinase A pathway. This has led to suggestions that NF1 has distinct Ras- and cAMP-related functions. Here we report that the Drosophila NF1 growth defect reflects a non-cell-autonomous requirement for NF1 in larval neurons that express the R-Ras ortholog Ras2, that NF1 is a GAP for Ras1 and Ras2, and that a functional NF1-GAP catalytic domain is both necessary and sufficient for rescue. Moreover, a Drosophila p120RasGAP ortholog, when expressed in the appropriate cells, can substitute for NF1 in growth regulation. Our results show that loss of NF1 can give rise to non-cell-autonomous developmental defects, implicate aberrant Ras-mediated signaling in larval neurons as the primary cause of the NF1 growth deficiency, and argue against the notion that neurofibromin has separable Ras- and cAMP-related functions. PMID- 17114579 TI - Rhomboid proteins: conserved membrane proteases with divergent biological functions. AB - The rhomboid gene was discovered in Drosophila, where it encodes a seven transmembrane protein that is the signal-generating component of epidermal growth factor (EGF) receptor signaling during development. Although metazoan developmental regulators are rarely conserved outside the animal kingdom, rhomboid proteins are conserved in all kingdoms of life, but the significance of this remains unclear. Recent biochemical reconstitution and high-resolution crystal structures have provided proof that rhomboid proteins function as novel intramembrane proteases, with a serine protease-like catalytic apparatus embedded within the membrane bilayer, buried in a hydrophilic cavity formed by a protein ring. A thorough consideration of all known examples of rhomboid function suggests that, despite biochemical similarity in mechanism and specificity, rhomboid proteins function in diverse processes including quorum sensing in bacteria, mitochondrial membrane fusion, apoptosis, and stem cell differentiation in eukaryotes; rhomboid proteins are also now starting to be linked to human disease, including early-onset blindness, diabetes, and parasitic diseases. Regulating cell signaling is at the heart of rhomboid protein function in many, but not all, of these processes. Further study of these novel enzymes promises to reveal the evolutionary path of rhomboid protein function, which could provide insights into the forces that drive the molecular evolution of regulatory mechanisms. PMID- 17114580 TI - Precise destruction: an emerging picture of the APC. AB - Cell cycle transitions are often accompanied by the degradation of regulatory molecules. Targeting proteins to the proteasome for degradation is accomplished by the covalent addition of ubiquitin chains. The specificity of this pathway is largely dictated by a set of enzymes called ubiquitin ligases (or E3s). The anaphase-promoting complex (or APC) is a ubiquitin ligase that has a particularly prominent role in regulating cell cycle progression. To date, the APC is the most complicated member of the RING/cullin family of multisubunit E3s. It includes at least 13 core subunits and three related adaptors. A combination of biochemical, genetic, and structural approaches are now shedding light on the enzymology of the APC. This review will focus on these data, drawing parallels with related ubiquitin ligases. PMID- 17114581 TI - Variation in the epigenetic silencing of FLC contributes to natural variation in Arabidopsis vernalization response. AB - Vernalization, the cold-induced acceleration of flowering, involves the epigenetic silencing of the floral repressor gene FLOWERING LOCUS C (FLC). We investigated the molecular basis for variation in vernalization in Arabidopsis natural accessions adapted to different climates. A major variable was the degree to which different periods of cold caused stable FLC silencing. In accessions requiring long vernalization, FLC expression was reactivated following nonsaturating vernalization, but this reactivation was progressively attenuated with increasing cold exposure. This response was correlated with the rate of accumulation of FLC histone H3 Lys 27 trimethylation (H3K27me3). Thus, variation in epigenetic silencing of FLC appears to have contributed to Arabidopsis adaptation. PMID- 17114582 TI - MtHAP2-1 is a key transcriptional regulator of symbiotic nodule development regulated by microRNA169 in Medicago truncatula. AB - In the model legume Medicago truncatula, we identified a new transcription factor of the CCAAT-binding family, MtHAP2-1, for which RNA interference (RNAi) and in situ hybridization experiments indicate a key role during nodule development, possibly by controlling nodule meristem function. We could also show that MtHAP2 1 is regulated by microRNA169, whose overexpression leads to the same nodule developmental block as MtHAP2-1 RNAi constructs. The complementary expression pattern of miR169 and MtHAP2-1 and the phenotype of miR169-resistant MtHAP2-1 nodules strongly suggest, in addition, that the miR169-mediated restriction of MtHAP2-1 expression to the nodule meristematic zone is essential for the differentiation of nodule cells. PMID- 17114583 TI - The S. cerevisiae Rrm3p DNA helicase moves with the replication fork and affects replication of all yeast chromosomes. AB - The Saccharomyces cerevisiae DNA helicase Rrm3p is needed for normal fork progression through >1000 discrete sites scattered throughout the genome. Here we show that replication of all yeast chromosomes was markedly delayed in rrm3 cells. Delayed replication was seen even in a region that lacks any predicted Rrm3p-dependent sites. Based on the pattern of replication intermediates in two dimensional gels, the rate of fork movement in rrm3 cells appeared similar to wild-type except at known Rrm3p-dependent sites. These data suggest that although Rrm3p has a global role in DNA replication, its activity is needed only or primarily at specific, difficult-to-replicate sites. By the criterion of chromatin immunoprecipitation, Rrm3p was associated with both Rrm3p-dependent and -independent sites, and moved with the replication fork through both. In addition, Rrm3p interacted with Pol2p, the catalytic subunit of DNA polymerase epsilon, in vivo. Thus, rather than being recruited to its sites of action when replication forks stall at these sites, Rrm3p is likely a component of the replication fork apparatus. PMID- 17114584 TI - Smad4 is dispensable for normal pancreas development yet critical in progression and tumor biology of pancreas cancer. AB - SMAD4 is inactivated in the majority of pancreatic ductal adenocarcinomas (PDAC) with concurrent mutational inactivation of the INK4A/ARF tumor suppressor locus and activation of the KRAS oncogene. Here, using genetically engineered mice, we determined the impact of SMAD4 deficiency on the development of the pancreas and on the initiation and/or progression of PDAC-alone or in combination with PDAC- relevant mutations. Selective SMAD4 deletion in the pancreatic epithelium had no discernable impact on pancreatic development or physiology. However, when combined with the activated KRAS(G12D) allele, SMAD4 deficiency enabled rapid progression of KRAS(G12D)-initiated neoplasms. While KRAS(G12D) alone elicited premalignant pancreatic intraepithelial neoplasia (PanIN) that progressed slowly to carcinoma, the combination of KRAS(G12D) and SMAD4 deficiency resulted in the rapid development of tumors resembling intraductal papillary mucinous neoplasia (IPMN), a precursor to PDAC in humans. SMAD4 deficiency also accelerated PDAC development of KRAS(G12D) INK4A/ARF heterozygous mice and altered the tumor phenotype; while tumors with intact SMAD4 frequently exhibited epithelial-to mesenchymal transition (EMT), PDAC null for SMAD4 retained a differentiated histopathology with increased expression of epithelial markers. SMAD4 status in PDAC cell lines was associated with differential responses to transforming growth factor-beta (TGF-beta) in vitro with a subset of SMAD4 wild-type lines showing prominent TGF-beta-induced proliferation and migration. These results provide genetic confirmation that SMAD4 is a PDAC tumor suppressor, functioning to block the progression of KRAS(G12D)-initiated neoplasms, whereas in a subset of advanced tumors, intact SMAD4 facilitates EMT and TGF-beta-dependent growth. PMID- 17114585 TI - Aggressive pancreatic ductal adenocarcinoma in mice caused by pancreas-specific blockade of transforming growth factor-beta signaling in cooperation with active Kras expression. AB - Pancreatic ductal adenocarcinoma (PDAC) is an almost uniformly lethal disease in humans. Transforming growth factor-beta (TGF-beta) signaling plays an important role in PDAC progression, as indicated by the fact that Smad4, which encodes a central signal mediator downstream from TGF-beta, is deleted or mutated in 55% and the type II TGF-beta receptor (Tgfbr2) gene is altered in a smaller subset of human PDAC. Pancreas-specific Tgfbr2 knockout mice have been generated, alone or in the context of active Kras (Kras(G12D)) expression, using the Cre-loxP system driven by the endogenous Ptf1a (pancreatic transcription factor-1a) locus. Pancreas-selective Tgfbr2 knockout alone gave no discernable phenotype in 1.5 yr. Pancreas-specific Kras(G12D) activation alone essentially generated only intraepithelial neoplasia within 1 yr. In contrast, the Tgfbr2 knockout combined with Kras(G12D) expression developed well-differentiated PDAC with 100% penetrance and a median survival of 59 d. Heterozygous deletion of Tgfbr2 with Kras(G12D) expression also developed PDAC, which indicated a haploinsufficiency of TGF-beta signaling in this genetic context. The clinical and histopathological manifestations of the combined Kras(G12D) expression and Tgfbr2 knockout mice recapitulated human PDAC. The data show that blockade of TGF-beta signaling and activated Ras signaling cooperate to promote PDAC progression. PMID- 17114586 TI - Hedgehog/Ras interactions regulate early stages of pancreatic cancer. AB - Pancreatic ductal adenocarcinoma (PDA) constitutes a lethal disease that affects >30,000 people annually in the United States. Deregulation of Hedgehog signaling has been implicated in the pathogenesis of PDA. To gain insights into the role of the pathway during the distinct stages of pancreatic carcinogenesis, we established a mouse model in which Hedgehog signaling is activated specifically in the pancreatic epithelium. Transgenic mice survived to adulthood and developed undifferentiated carcinoma, indicating that epithelium-specific Hedgehog signaling is sufficient to drive pancreatic neoplasia but does not recapitulate human pancreatic carcinogenesis. In contrast, simultaneous activation of Ras and Hedgehog signaling caused extensive formation of pancreatic intraepithelial neoplasias, the earliest stages of human PDA tumorigenesis, and accelerated lethality. These results indicate the cooperation of Hedgehog and Ras signaling during the earliest stages of PDA formation. They also mark Hedgehog pathway components as relevant therapeutic targets for both early and advanced stages of pancreatic ductal neoplasia. PMID- 17114587 TI - p63 regulates proliferation and differentiation of developmentally mature keratinocytes. AB - p63 is a multi-isoform p53 family member required for epidermal development. Contrasting roles for p63 in either the initial commitment to the stratified epithelial cell fate or in stem cell-based self-renewal have been proposed. To investigate p63 function in a post-developmental context, we used siRNAs directed against p63 to down-regulate p63 expression in regenerating human epidermis. Loss of p63 resulted in severe tissue hypoplasia and inhibited both stratification and differentiation in a cell-autonomous manner. Although p63-deficient cells exhibited hypoproliferation, differentiation defects were not due to tissue hypoplasia. Simultaneous p63 and p53 knockdown rescued the cell proliferation defect of p63 knockdown alone but failed to restore differentiation, suggesting that defects in epidermal proliferation and differentiation are mediated via p53 dependent and -independent mechanisms, respectively. Furthermore, DeltaNp63 isoforms are the main mediators of p63 effects, although TAp63 isoforms may contribute to late differentiation. These data indicate that p63 is required for both the proliferative and differentiation potential of developmentally mature keratinocytes. PMID- 17114588 TI - Trachoma, antibiotics and randomised controlled trials. PMID- 17114589 TI - Rebound tonometry: new opportunities and limitations of non-invasive determination of intraocular pressure. PMID- 17114590 TI - Vascular endothelial growth factor biology: clinical implications for ocular treatments. AB - Decades of research on vascular endothelial growth factor (VEGF) have reached fruition with the recent development of intravitreal anti-VEGF treatments for exudative age-related macular degeneration. VEGF is a critical regulator of angiogenesis and vascular permeability with diverse roles, both pathological and physiological, during development and adulthood. The aim of this article is to review aspects of VEGF biology that may be relevant to the clinical use of anti VEGF agents in ophthalmology: molecular characteristics and isoforms of VEGF; its roles in vasculogenesis, vascular maintenance and angiogenesis; systemic effects of VEGF inhibition; and properties of current anti-VEGF agents. PMID- 17114591 TI - Nyctalopia and hemeralopia: the current usage trend in the literature. PMID- 17114592 TI - Lothian combined paediatric ophthalmology and rheumatology service. PMID- 17114593 TI - A tiger with glaucoma. PMID- 17114594 TI - Buckle, no cryo: scleral buckle with no cryotherapy for retinal detachment secondary to commotio retinae. PMID- 17114595 TI - Validity and reliability of the classroom participation questionnaire with deaf and hard of hearing students in public schools. AB - The Classroom Participation Questionnaire (CPQ) was administered to 136 deaf or hard of hearing (D/HH) students attending general education classrooms in Grades 4-10. The CPQ is a student-rated measure that yields scores for Understanding Teachers, Understanding Students, Positive Affect, and Negative Affect. Validity and reliability of a long (28-item) and a short (16-item) form are reported. We provide evidence of (a) internal structure validity through an examination of the relationships between the subscales and an analysis of interitem reliability within each scale, (b) reliability over time by examining the scores of students over a 3-year period, and (c) external structure validity through an examination of the relationships of the CPQ with measures of teacher-rated academic competence and Stanford achievement scores. The results suggest that both the long and short form of the CPQ can be used to assess participation of D/HH students in general education classrooms. PMID- 17114596 TI - Radial mass density, charge, and epitope distribution in the Cryptococcus neoformans capsule. AB - Exposure of Cryptococcus neoformans cells to gamma radiation results in a gradual release of capsular polysaccharide, in a dose-dependent manner. This method allows the systematic exploration of different capsular regions. Using this methodology, capsule density was determined to change according to the radial distribution of glucuronoxylomannan and total polysaccharide, becoming denser at the inner regions of the capsule. Scanning electron microscopy of cells following gamma radiation treatment confirmed this finding. The zeta potential of the capsule also increased as the capsule size decreased. However, neither charge nor density differences were correlated with any change in sugar composition (xylose, mannose, and glucuronic acid) in the different capsular regions, since the proportions of these sugars remained constant throughout the capsule. Analysis of the capsular antigenic properties by monoclonal antibody binding and Scatchard analysis revealed fluctuations in the binding affinity within the capsule but not in the number of antibody binding sites, suggesting that the spatial organization of high- and low-affinity epitopes within the capsule changed according to radial position. Finally, evidence is presented that the structure of the capsule changes with capsule age, since the capsule of older cells became more resistant to gamma radiation-induced ablation. In summary, the capsule of C. neoformans is heterogeneous in its spatial distribution and changes with age. Furthermore, our results suggest several mechanisms by which the capsule may protect the fungal cell against exogenous environmental factors. PMID- 17114597 TI - Ste12 transcription factor homologue CpST12 is down-regulated by hypovirus infection and required for virulence and female fertility of the chestnut blight fungus Cryphonectria parasitica. AB - A putative homologue of the Saccharomyces cerevisiae Ste12 transcription factor was identified in a series of expressed sequence tag-based microarray analyses as being down-regulated in strains of the chestnut blight fungus, Cryphonectria parasitica, infected by virulence-attenuating hypoviruses. Cloning of the corresponding gene, cpst12, confirmed a high level of similarity to Ste12 homologues of other filamentous fungi. Disruption of cpst12 resulted in no alterations in in vitro growth characteristics or colony morphology and an increase in the production of asexual spores, indicating that CpST12 is dispensable for vegetative growth and conidiation on artificial medium. However, the disruption mutants showed a very substantial reduction in virulence on chestnut tissue and a complete loss of female fertility, two symptoms normally conferred by hypovirus infection. Both virulence and female fertility were restored by complementation with the wild-type cpst12 gene. Analysis of transcriptional changes caused by cpst12 gene disruption with a custom C. parastica cDNA microaray chip identified 152 responsive genes. A significant number of these putative CpST12-regulated genes were also responsive to hypovirus infection. Thus, cpst12 encodes a cellular transcription factor, CpST12, that is down-regulated by hypovirus infection and required for female fertility, virulence and regulated expression of a subset of hypovirus responsive host genes. PMID- 17114598 TI - Vesicular polysaccharide export in Cryptococcus neoformans is a eukaryotic solution to the problem of fungal trans-cell wall transport. AB - The mechanisms by which macromolecules are transported through the cell wall of fungi are not known. A central question in the biology of Cryptococcus neoformans, the causative agent of cryptococcosis, is the mechanism by which capsular polysaccharide synthesized inside the cell is exported to the extracellular environment for capsule assembly and release. We demonstrate that C. neoformans produces extracellular vesicles during in vitro growth and animal infection. Vesicular compartments, which are transferred to the extracellular space by cell wall passage, contain glucuronoxylomannan (GXM), a component of the cryptococcal capsule, and key lipids, such as glucosylceramide and sterols. A correlation between GXM-containing vesicles and capsule expression was observed. The results imply a novel mechanism for the release of the major virulence factor of C. neoformans whereby polysaccharide packaged in lipid vesicles crosses the cell wall and the capsule network to reach the extracellular environment. PMID- 17114599 TI - Treatment of toenail onychomycosis with oral terbinafine plus aggressive debridement: IRON-CLAD, a large, randomized, open-label, multicenter trial. AB - This study was conducted to investigate the efficacy of oral terbinafine with and without aggressive debridement for the treatment of toenail onychomycosis. Onychomycosis patients aged 18 to 75 years received 12 weeks of terbinafine, 250 mg/day, alone (n = 255) or with aggressive debridement (n = 249). Both groups showed marked improvement from baseline at all time points. At week 48, complete, mycologic, and clinical cure rates were higher in the terbinafine plus debridement group compared with the terbinafine alone group, although significance was reached only for clinical cure (59.8% versus 51.4%; P = .023). Although approximately 39% of the patients received at least one antidiabetic, antihypertensive, or cholesterol-lowering agent concomitantly, including statins, the incidence of treatment-emergent adverse events was low and the adverse events were generally mild to moderate in severity. No clinically significant changes in liver transaminase levels were observed 6 weeks after treatment or after 12 weeks in those tested. These results support the well-established safety and efficacy of terbinafine for treatment of onychomycosis. PMID- 17114600 TI - Effect of functional foot orthoses on first metatarsophalangeal joint dorsiflexion in stance and gait. AB - Reduction in first metatarsophalangeal joint maximum degree of dorsiflexion with dorsiflexion of the first ray has been proposed to be the predominant cause of hallux abducto valgus and hallux rigidus. We sought to determine whether orthoses made from a cast with the first ray plantarflexed and a 4-mm medial skive could increase the maximum degree of dorsiflexion in patients with functional hallux limitus in stance and gait. Forty-eight feet of 27 subjects were casted for orthoses with the first ray plantarflexed and in the customary neutral rearfoot position with locked midtarsal joint. First metatarsophalangeal joint maximum dorsiflexion was measured with and without orthoses in stance, and subhallux pressure was measured with and without orthoses at heel-off. Changes in mean maximum dorsiflexion in stance and in mean maximum subhallux pressure in gait with orthoses were significant. We investigated the relationship between this increase in dorsiflexion and gender, shoe size, resting calcaneal stance position, and change in resting calcaneal stance position with the use of orthoses. These correlations were not statistically significant. The biomechanical implication of increasing limited first metatarsophalangeal joint dorsiflexion with orthoses is discussed and related to the clinical treatment of deformities, including hallux valgus and hallux rigidus. The use of orthoses to decrease subhallux pressure is also discussed. PMID- 17114601 TI - Efficacy of concentrated autologous platelet-derived growth factors in chronic lower-extremity wounds. AB - The efficacy of concentrated autologous platelet-derived growth factors in the healing and closure of chronic lower-extremity wounds was evaluated in 24 patients with 33 lower-extremity wounds treated previously for at least 6 months using traditional methods. Surgical wound debridement was performed to convert chronic ulcers into acute wounds. Concentrated autologous platelet-derived growth factors and thrombin were applied to the wound bases and protected with a nonadhering compression dressing that remained intact for 7 days. Wounds were evaluated and the concentrate was reapplied every 2 weeks. Wound closure and complete epithelialization was achieved in 20 wounds. Seventy-five percent or greater wound closure was obtained in three wounds, 50% to 74% closure in three wounds, and 25% to 49% closure in two wounds. Five wounds displayed no improvement. Mean time to complete closure was 11.15 weeks. The application of concentrated autologous platelet-derived growth factors and thrombin resulted in substantial wound healing and wound-diameter reduction. This technique constitutes a safe and effective treatment option and avoids lengthy treatment periods that increase the potential for infection. PMID- 17114602 TI - Use of plantar contact area to predict medial longitudinal arch height during walking. AB - A study was conducted to determine whether plantar surface contact area measures calculated from footprints collected during walking can be used to predict the height of the medial longitudinal arch. Thirty healthy women participated in the study. Arch height was determined by the distance from the navicular tuberosity to the floor and by the "bony" arch index. Dynamic plantar surface contact area was recorded using a pressure platform as the subjects walked across a 12-m walkway. The arch index and the total plantar surface contact area were determined from the pressure sensor data. The results indicated that plantar surface contact area could be used to estimate only approximately 27% of the height of the medial longitudinal arch as determined by navicular tuberosity height and the bony arch index. These findings demonstrate the inability of the clinician to predict the vertical height of the medial longitudinal arch on the basis of the amount of foot plantar surface area in contact with the ground during walking. PMID- 17114603 TI - The biomechanical effects of talectomy on the foot. AB - The biomechanical effects of talectomy on the foot were investigated in seven fresh below-the-knee amputation specimens using pressure-sensitive films placed on the facets of the calcaneus, footprints, and loading-pattern diagrams in the intact foot and after talectomy with anterior and posterior displacement of the foot. Both talectomy techniques distorted the loads carried by the facets of the calcaneus. In the intact foot, 65.6% of the loads were carried by the posterior facet of the calcaneus and 34.4% by the anterior and middle facets. After talectomy with anterior displacement of the foot, although the loads carried by the anterior and middle facets decreased significantly (P = .018), the increase in the loads carried by the posterior facet was not significant compared with the intact foot (P = .176). Similarly, the loads carried by the posterior facet decreased significantly after talectomy with posterior displacement of the foot (P = .028), but the increases in the loads carried by the anterior and middle facets were not significant (P = .735). Comparing the two types of talectomy, the loads carried by each facet changed significantly (P = .018). Talectomy with posterior displacement of the foot also changed the loading patterns and resulted in significant pronation of the foot. These results suggest that talectomy should be performed only as a salvage procedure and that talectomy with anterior displacement of the foot may be preferred when talectomy is indicated. PMID- 17114604 TI - Relationship of functional leg-length discrepancy to abnormal pronation. AB - The objective of this study was to determine whether a correlation exists between abnormal pronation and functional leg-length discrepancies. Visual assessment and a pelvic thrust maneuver were used to identify the functionally short leg in 56 indigenous Mexicans (20 males and 36 females; mean age, 33 years; mean weight, 59 kg; and mean height, 1.60 m). The Foot Posture Index was used with a modified stance position to identify the more pronated foot. The posterosuperior iliac spines were used to identify the "relative" position of the innominate bones. The raw data obtained from this study were evaluated using the McNemar test for paired proportions. A significant positive correlation was found between abnormal pronation and hip position and between hip position and functional leg-length discrepancy. These results are consistent with a theoretical ascending dysfunctional pelvic model: Abnormal pronation pulls the innominate bones anteriorly (forward); anterior rotation of the innominate bones shifts the acetabula posteriorly and cephalad (backward and upward); and this shift in the acetabula hyperextends the knees and shortens the legs, with the shortest leg corresponding to the most pronated foot. PMID- 17114607 TI - Arthroplasty of the interphalangeal joint of the great toe using costal osteochondral grafting. AB - Although many reports have been published on the usefulness of costal cartilage grafting in the reconstruction of interphalangeal joints of fingers, there are only a few published reports on the reconstruction of interphalangeal joints of toes. We describe a 21-year-old woman with a tissue defect of the dorsum pedis and a partial defect of the interphalangeal joint of the great toe caused by a motor-vehicle accident. We attempted arthroplasty using a free latissimus dorsi myocutaneous flap and a costal osteochondral graft. The grafted rib and cartilage survived, allowing the patient to resume functional ambulation for day-to-day activities. Arthroplasty using costal osteochondral grafts seems to be an effective means of reconstructing the interphalangeal joints of toes. PMID- 17114608 TI - Desyndactylization of the first and second toes using full-thickness autologous skin graft from the ankle. AB - We present a unique case of congenital bilateral simple syndactyly of the first and second toes that was surgically treated using a full-thickness skin graft harvested from the same foot at the lateral aspect of the ankle. This surgical approach eliminates the potential need to involve another surgical team to harvest a donor graft from above the ankle, saving operating room time, anesthesia time, and overall cost to the patient. Cosmetically, scar formation above the ankle is also eliminated. PMID- 17114609 TI - Virtual microscopes in podiatric medical education. AB - In many medical schools, microscopes are being replaced as teaching tools by computers with software that emulates the use of a light microscope. This article chronicles the adoption of "virtual microscopes" by a podiatric medical school and presents the results of educational research on the effectiveness of this adoption in a histology course. If the trend toward virtual microscopy in education continues, many 21st-century physicians will not be trained to operate a light microscope. The replacement of old technologies by new is discussed. The fundamental question is whether all podiatric physicians should be trained in the use of a particular tool or only those who are likely to use it in their own practice. PMID- 17114610 TI - Cutaneous ulceration secondary to hydroxyurea treatment. PMID- 17114613 TI - Has the time arrived to image placental perfusion? AB - Taillieu et al have shown that it is possible to noninvasively measure the placental blood flow, fractional volume of the maternal vascular placental compartment, and rate of transfer of contrast material between the maternal and fetal circulation in gravid mice through the use of dynamic contrast-enhanced MR imaging. Clearly, much work lies ahead before functional evaluation of the placenta becomes a clinical reality, but contrast-enhanced imaging shows promise for functional evaluation of placental disease. PMID- 17114614 TI - Building research programs in diagnostic radiology. Part I. Framing the issues. PMID- 17114615 TI - RSNA Clinical Trials Methodology Workshop. PMID- 17114616 TI - RSNA takes another step in support of imaging research. PMID- 17114617 TI - Standardized nomenclature and description of CT scanning techniques. PMID- 17114618 TI - Standardization in CT terminology: a physicist's perspective. PMID- 17114619 TI - Recent advances in chest radiography. AB - There have been many remarkable advances in conventional thoracic imaging over the past decade. Perhaps the most remarkable is the rapid conversion from film based to digital radiographic systems. Computed radiography is now the preferred imaging modality for bedside chest imaging. Direct radiography is rapidly replacing film-based chest units for in-department posteroanterior and lateral examinations. An exciting aspect of the conversion to digital radiography is the ability to enhance the diagnostic capabilities and influence of chest radiography. Opportunities for direct computer-aided detection of various lesions may enhance the radiologist's accuracy and improve efficiency. Newer techniques such as dual-energy and temporal subtraction radiography show promise for improved detection of subtle and often obscured or overlooked lung lesions. Digital tomosynthesis is a particularly promising technique that allows reconstruction of multisection images from a short acquisition at very low patient dose. Preliminary data suggest that, compared with conventional radiography, tomosynthesis may also improve detection of subtle lung lesions. The ultimate influence of these new technologies will, of course, depend on the outcome of rigorous scientific validation. PMID- 17114620 TI - Computer-aided detection in digital mammography: comparison of craniocaudal, mediolateral oblique, and mediolateral views. AB - PURPOSE: To retrospectively compare the sensitivity of a computer-aided detection (CAD) system for depicting breast cancer in three digital mammographic views. MATERIALS AND METHODS: This study was conducted with institutional review board approval; informed consent was waived. A commercially available CAD system was applied to the craniocaudal, mediolateral oblique, and mediolateral digital mammographic views of 83 women (mean age, 48 years; range, 30-66 years) with 83 histologically proved breast cancers. Findings were 59 masses and 41 microcalcifications (17 lesions showed both findings; 42 lesions, mass only; and 24 lesions, microcalcification only). The paired t test was used to analyze sensitivity of the CAD system for the detection of cancer in these three mammographic views and in combinations of the views. RESULTS: The sensitivities of the CAD system were 92% (76 of 83) in the craniocaudal view, 83% (69 of 83) in the mediolateral oblique view, and 86% (71 of 83) in the mediolateral view; the differences were not significant (P = .07-.62). Sensitivity increased to 96% (80 of 83) in the craniocaudal plus mediolateral oblique views and to 99% (82 of 83) in the craniocaudal plus mediolateral oblique plus mediolateral views. For masses, the sensitivity of the CAD system was 76% (45 of 59) in the craniocaudal view and 75% (44 of 59) in the mediolateral oblique view and increased to 93% (55 of 59) when mediolateral oblique and craniocaudal views were combined (P < .001). For microcalcifications, sensitivity was 98% (40 of 41) in the craniocaudal view and 95% (39 of 41) in the mediolateral oblique view, and this increased to 100% (41 of 41) when the mediolateral oblique and craniocaudal views were combined (P = .31). CONCLUSION: The sensitivities of the CAD system were not significantly different among these three digital mammographic views. Sensitivity for depicting masses was significantly increased (P < .001) when the craniocaudal view was added to the mediolateral oblique view. PMID- 17114621 TI - Effects of surgical ventricular restoration on left ventricular function: dynamic MR imaging. AB - PURPOSE: To retrospectively evaluate with dynamic magnetic resonance (MR) imaging the changes in global and regional left ventricular (LV) function after surgical ventricular restoration (SVR) performed in chronic ischemic heart disease patients with large nonaneurysmal or aneurysmal postmyocardial infarction zones. MATERIALS AND METHODS: The study was performed with institutional review board approval, and a waiver of individual informed consent was obtained. The study was HIPAA compliant. Patients (83 men, 22 women; mean age, 61 years +/- 9 [standard deviation]) were evaluated with MR imaging before and after SVR as follows: pre SVR examination (n = 105; 25 days +/- 39 before SVR; median, 7 days; range, 1-189 days), early post-SVR examination (n = 95, 7 days +/- 3 after SVR), and late post SVR (n = 35, 313 days +/- 158 after SVR). Cine MR imaging allowed calculation of ejection fraction and rate-corrected velocity of circumferential fiber shortening (Vcf(C)) for global LV functional evaluation, whereas tagged MR imaging (spatial modulation of magnetization with harmonic phase analysis) permitted assessment of regional circumferential strain (E(C)) with coronary distribution. Vcf(C) and E(C) were computed at both LV base- and mid-LV short-axis levels remote from the site of anteroapical SVR. RESULTS: Prior to SVR, LV dilatation and diminished global and regional LV function were observed. At early post-SVR examination, Vcf(C) had improved significantly but E(C) showed a worsening trend overall, although only E(C )of the right coronary artery at the mid-LV level worsened significantly. At late post-SVR examination, Vcf(C) values were improved when compared with pre-SVR values, although E(C) showed no statistically significant improvement. When compared with that at early post-SVR examination, however, E(C) showed significant improvement in two segments: left anterior descending artery and right coronary artery at mid-LV level. CONCLUSION: Although volume-based indexes of global LV function improve significantly after SVR, regional LV function did not improve significantly; there was evidence of continued LV remodeling after SVR. PMID- 17114622 TI - CT of small-bowel ischemia associated with obstruction in emergency department patients: diagnostic performance evaluation. AB - PURPOSE: To retrospectively evaluate the diagnostic performance of computed tomography (CT) for detection of small-bowel ischemia in emergency department patients with abdominal pain and to compare the prospective interpretation with a retrospective interpretation by using surgical or pathologic findings as the reference standard. MATERIALS AND METHODS: The HIPAA-compliant study was approved by the institutional review board, and patients consented to research authorization. Sixty patients (61 examinations) (25 male, 35 female patients; median age, 67 years; range, 0.9-89.7 years) with acute abdominal pain underwent immediate abdominal and pelvic CT and subsequent surgery of the small bowel within 7 days of CT. Prospective radiologic reports were reviewed for diagnosis of small-bowel obstruction and ischemia. Two gastrointestinal radiologists performed blinded, independent, retrospective review of the CT studies with no clinical data other than presence of acute abdominal pain. The reviewers categorized CT signs of obstruction and ischemia and estimated diagnostic certainty. Discordant findings were resolved by consensus review by a third gastrointestinal radiologist. CT interpretations were compared with prospective interpretations and surgical or pathologic findings. Sensitivity and specificity estimates with confidence intervals were calculated. Fisher exact and chi2 tests were used to assess associations between CT signs and the diagnosis of ischemia; kappa statistics were used to estimate agreement between readers. RESULTS: In 27 (44%) of 61 CT studies, small-bowel ischemia was surgically or pathologically confirmed. Sensitivity and specificity for the diagnosis of ischemia were, respectively, 14.8% and 94.1% for prospective interpretations, 29.6% and 91.2% for reader 1, 40.7% and 85.3% for reader 2, and 51.9% and 88.2% for the consensus review. Decreased segmental enhancement was the most specific sign for small bowel ischemia (P = .001), and its recognition would have improved the diagnostic performance of all readers. There was a significant association of the small bowel feces sign with the presence of small-bowel ischemia (P = .046). CONCLUSION: Diagnostic performance assessment of CT for the diagnosis of small bowel ischemia revealed poor prospective interpretation sensitivity. PMID- 17114623 TI - MR-guided radiofrequency ablation: do magnetic fields influence extent of coagulation in ex vivo bovine livers? AB - PURPOSE: To prospectively determine if static magnetic fields of magnetic resonance (MR) imagers affect radiofrequency (RF) ablation coagulation volume and shape. MATERIALS AND METHODS: Ex vivo RF ablations of bovine livers were performed with magnetic field strengths of 0.2, 1.5, and 3.0 T and were compared with ablations performed outside the magnetic field in a control group. Two MR compatible monopolar RF devices (internally cooled single and cluster electrodes) were systematically tested. Length of long axis (y-axis), length of two short axes (x- and z-axes), and coagulation volume and shape measured outside and inside different magnetic fields were compared with the Dunnett test. Significance level was set to .05. RESULTS: For the single electrode, no significant difference was observed between length of short axes and coagulation volume and shape measured inside and outside the magnetic field. Mean x- and z axis lengths were 2.3 and 2.6 cm, respectively, outside the magnetic field; 2.4 and 2.4 cm, respectively, at 0.2 T; 2.5 and 2.6 cm, respectively, at 1.5 T; and 2.2 and 2.5 cm, respectively, at 3.0 T. Differences between length of long axis, length of short axis perpendicular to static magnetic field, and coagulation volume and shape achieved with the cluster electrode inside and outside the magnetic field were not significant. Mean x- and z-axis lengths were 3.9 and 3.9 cm, respectively, outside the magnetic field; 3.7 and 3.8 cm, respectively, at 0.2 T; 4.0 and 4.3 cm, respectively, at 1.5 T; and 3.8 and 3.8 cm, respectively, at 3.0 T. Differences between ablations performed at 1.5 T and those performed in the control group with the cluster electrode were significant (P = .026). In this case, a difference of 4 mm in the length of the short axis parallel to the magnetic field was detected, but there was no significant difference in coagulation volume. CONCLUSION: No significant differences in coagulation volume and shape could be recorded between RF ablations performed outside and those performed inside the static magnetic field. PMID- 17114624 TI - Elastase-induced pulmonary emphysema in rats: comparison of computed density and microscopic morphometry. AB - PURPOSE: To prospectively compare computed tomographic (CT) quantification of pulmonary emphysema in elastase-treated rats with morphometry and to evaluate the information yielded by CT quantification and pulmonary function tests. MATERIALS AND METHODS: This study was approved by the local committee on care and use of animals in research. Thirty-six animals were used. Emphysema was produced by means of one or two tracheal injections of 300 IU of elastase, 8 weeks apart, in seven and 12 rats, respectively. As a control group, 10 rats received an injection of normal saline. The dynamic resistance, dynamic compliance, and static compliance were measured. CT was performed with 1-mm section thickness and 3-mm intervals. Relative areas of lung with attenuation coefficients lower than nine chosen thresholds (from -900 to -980 HU) and eight percentiles (from 1st to 18th percentiles) of the distribution of attenuation coefficients were compared with measurements of alveoli size--that is, mean interwall distance (MIWD) and mean perimeter per field (MP). Correlations between data obtained with thresholds and percentiles and MIWD and MP were investigated by means of Spearman coefficients (r(s)). Values of pulmonary function tests, most appropriate relative area threshold, and percentile were investigated by means of stepwise multiple regressions. RESULTS: For thresholds, relative surface area with attenuation coefficients less than -940 HU (RA(940)) showed the strongest correlations with findings at microscopy (r(s) = 0.676, P < .001 for MIWD; r(s) = -0.720, P < .001 for MP). For percentiles, the 3rd percentile showed the strongest correlations (r(s) = -0.647, P < .001 for MIWD; r(s) = 0.701, P < .001 for MP). Dynamic compliance and RA(940) or 3rd percentile were complementary for predicting microscopic measurements. CONCLUSION: In rats, RA(940) and the 3rd percentile reflect the extent of elastase-induced pulmonary emphysema and are complementary to dynamic compliance to predict microscopic extent. PMID- 17114625 TI - Vascular access: comparison of US guidance with the sonic flashlight and conventional US in phantoms. AB - PURPOSE: To prospectively evaluate whether ultrasonography (US)-guided vascular access can be learned and performed faster with the sonic flashlight than with conventional US and to demonstrate sonic flashlight-guided vascular access in a cadaver. MATERIALS AND METHODS: Institutional review board approval and oral and written informed consent were obtained. The sonic flashlight replaces the standard US monitor with a real-time US image that appears to float beneath the skin and is displayed where it is scanned. In studies 1 and 2, participants performed sonic flashlight-guided needle insertion tasks in vascular phantoms. In study 1, 16 participants (nine women, seven men) with no US experience performed 60 simulated vascular access trials with sonic flashlight or conventional US guidance. With analysis of variance (ANOVA) and power-curve fitting, improvement with practice rate and mean differences between techniques and tasks were examined. In study 2, 14 female nurses (mean age, 50.1 years) proficient with conventional US performed simulated vascular access trials on three tasks with the sonic flashlight and conventional US. With random assignment, half the participants used the sonic flashlight first and half used conventional US first. Mean performance with each technique and that with each task were compared by using ANOVA. In study 3, feasibility of sonic flashlight guidance for access to internal jugular and basilic veins was demonstrated in a cadaver. RESULTS: For study 1, learning rates (ie, decrease in access time over trials) did not differ for vascular access with sonic flashlight and conventional US. Overall, participants achieved faster vascular access times with sonic flashlight guidance (P < .007). In study 2, participants performed procedures faster overall with the sonic flashlight (P < .02) and found the sonic flashlight easier to use. In study 3, sonic flashlight-guided vascular access was gained in the cadaver. CONCLUSION: Learning and performance of vascular access were significantly faster with the sonic flashlight than with conventional US, and vascular access could be gained in a cadaver; the sonic flashlight is ready for clinical trials. PMID- 17114626 TI - Perforated versus nonperforated acute appendicitis: accuracy of multidetector CT detection. AB - PURPOSE: To retrospectively evaluate the accuracy of multidetector computed tomography (CT) in the diagnosis of perforated acute appendicitis by using surgery and pathologic examination combined as the reference standard. MATERIALS AND METHODS: The study was institutional review board approved and HIPAA compliant. Informed patient consent was waived. The authors retrospectively identified 244 patients (150 male, 94 female; mean age, 32.8 years; age range, 4 83 years) with pathologically proved acute appendicitis who underwent abdominopelvic multidetector CT. Two radiologists reviewed in consensus the multidetector CT images obtained in all patients for various findings that may be associated with appendiceal perforation. For continuous variables, a comparison of means between the perforated and nonperforated groups was performed by using the Wilcoxon rank sum test. For categorical variables, the sensitivity and specificity of each CT finding for the diagnosis of perforated appendicitis were determined. RESULTS: The CT findings of abscess (99%), extraluminal gas (98%), and ileus (93%) had the highest specificities for appendiceal perforation; however, the sensitivities of these findings were low: 34%, 35%, and 53%, respectively. The appendix was larger in patients with perforated appendicitis: The mean diameter was 15.1 mm compared with a mean diameter of 11.7 mm in patients with nonperforated appendicitis (P < .001). Appendicolith, free fluid, enlarged abdominal lymph nodes, and enhancement defect in the appendiceal wall were neither highly sensitive nor highly specific for the detection of perforation. CONCLUSION: Although certain multidetector CT findings are very specific for the diagnosis of perforated appendicitis, overall multidetector CT sensitivity is poor. Unless abscess or extraluminal gas is present, multidetector CT cannot enable the diagnosis of perforation. PMID- 17114627 TI - Polyps: linear and volumetric measurement at CT colonography. AB - PURPOSE: To retrospectively determine which of several computed tomographic (CT) colonography-based polyp measurements is most compatible with the linear measurement at optical colonoscopy and which is best for assessing change in polyp size. MATERIALS AND METHODS: This HIPAA-compliant study had institutional review board approval; informed consent was obtained. Prone and supine CT colonography with same-day optical colonoscopy was performed in 216 patients (147 men and 69 women; age range, 46-79 years; mean age, 59.2 years) with 338 polyps detected at CT colonography. Polyp size was measured with three linear measurements and two volume measurements. One linear measurement and one volume measurement were performed by using automated segmentation; remaining measurements were performed manually. Compatibility with linear size at optical colonoscopy and measurement reproducibility were assessed three ways: variation from size measurement at optical colonoscopy, change between prone and supine scans, and variability between observers. Confidence analysis assessed the ability of each measurement to identify polyps with an optical colonoscopy measurement of 1 cm or greater. RESULTS: Two hundred fifty-one segmentable polyps were present on both supine and prone scans. Linear polyp diameter manually measured on a three-dimensional endoluminally viewed surface (L(M3D)) indicated with 95% confidence that a polyp measured as 0.8 cm or smaller was less than 1.0 cm at optical colonoscopy. Prone and supine polyp size difference was smallest for L(M3D) and the linear diameter computed from manual and automated volume measurements, with interquartile ranges smaller than or equal to 0.3, 0.2, and 0.5 cm, respectively. Interobserver and intraobserver variability was smallest for linear polyp diameter measurements on a two-dimensional display, with a mean percentage difference of 2.8% (95% Bland-Altman limits of agreement: -17.8%, 23.4%) and 5.0% (95% Bland-Altman limits of agreement: -28.3%, 38.3%), respectively. CONCLUSION: L(M3D) best approximated polyp size measurements at optical colonoscopy. Linear diameter calculated from automated volume measurements showed the smallest variation between supine and prone scans while avoiding observer variability and may be best for assessing polyp size changes with serial examinations. PMID- 17114628 TI - Functional evaluation of transplanted kidneys with diffusion-weighted and BOLD MR imaging: initial experience. AB - PURPOSE: To prospectively evaluate feasibility and reproducibility of diffusion weighted (DW) and blood oxygenation level-dependent (BOLD) magnetic resonance (MR) imaging in patients with renal allografts, as compared with these features in healthy volunteers with native kidneys. MATERIALS AND METHODS: The local ethics committee approved the study protocol; patients provided written informed consent. Fifteen patients with a renal allograft and in stable condition (nine men, six women; age range, 20-67 years) and 15 age- and sex-matched healthy volunteers underwent DW and BOLD MR imaging. Seven patients with renal allografts were examined twice to assess reproducibility of results. DW MR imaging yielded a total apparent diffusion coefficient including diffusion and microperfusion (ADC(tot)), as well as an ADC reflecting predominantly pure diffusion (ADC(D)) and the perfusion fraction. R2* of BOLD MR imaging enabled the estimation of renal oxygenation. Statistical analysis was performed, and analysis of variance was used for repeated measurements. Coefficients of variation between and within subjects were calculated to assess reproducibility. RESULTS: In patients, ADC(tot), ADC(D), and perfusion fraction were similar in the cortex and medulla. In volunteers, values in the medulla were similar to those in the cortex and medulla of patients; however, values in the cortex were higher than those in the medulla (P < .05). Medullary R2* was higher than cortical R2* in patients (12.9 sec(-1) +/- 2.1 [standard deviation] vs 11.0 sec(-1) +/- 0.6, P < .007) and volunteers (15.3 sec(-1) +/- 1.1 vs 11.5 sec(-1) +/- 0.5, P < .0001). However, medullary R2* was lower in patients than in volunteers (P < .004). Increased medullary R2* was paralleled by decreased diffusion in patients with allografts. A low coefficient of variation in the cortex and medulla within subjects was obtained for ADC(tot), ADC(D), and R2* (<5.2%), while coefficient of variation within subjects was higher for perfusion fraction (medulla, 15.1%; cortex, 8.6%). Diffusion and perfusion indexes correlated significantly with serum creatinine concentrations. CONCLUSION: DW and BOLD MR imaging are feasible and reproducible in patients with renal allografts. PMID- 17114629 TI - Neural stem cell transplant survival in brains of mice: assessing the effect of immunity and ischemia by using real-time bioluminescent imaging. AB - PURPOSE: To use bioluminescent imaging in a murine transplant model to monitor the in vivo responses of transplanted luciferase-gene-positive neural progenitor cells (NPCs) to host immunity and ischemia. MATERIALS AND METHODS: All animal studies were conducted according to institutional guidelines, with approval of the Subcommittee on Research Animal Care. Cranial windows were created in all animals, and all animals underwent NPC (C17.2-Luc-GFP-gal) transplantation into the right basal ganglia. An observational study was performed on C57 BL/6 (n = 5), nude (n = 4), and CD-1 (n = 4) mice, with bioluminescent imaging performed at days 7, 11, and 14 after transplantation. A study on the effects of ischemia was performed in a similar manner, but with the following differences: On day 9 after transplantation, the C57 BL/6 mice underwent 18 minutes of transient forebrain ischemia by means of temporary bilateral carotid occlusions (n = 6). A control group of C57 BL/6 mice underwent sham surgery (n = 6). Bioluminescent imaging was performed on the ischemic animals and control animals at days 7, 9, 11, and 14. Repeated-measures analysis of variance or Student t test was used to compare the means of the luciferase activities. RESULTS: In vivo cell tracking demonstrated that (a) C17.2-Luc-GFP-gal NPCs survived and proliferated better in the T-cell deficient nude mice than in the immunocompetent C57 BL/6 or CD-1 mice, in which progressive immune mediated cell loss was shown, and (b) transient forebrain ischemia appeared, unexpectedly, to act as a short-term stimulus to transplanted NPC growth and survival in immunocompetent mice. CONCLUSION: Immune status and host immunity can have an influence on NPC graft survival, and these changes can be noninvasively assessed with bioluminescent imaging in this experimental model. PMID- 17114630 TI - Lung cancer: computerized quantification of tumor response--initial results. AB - PURPOSE: To prospectively quantify tumor response or progression in patients with lung cancer by using thin-section computed tomography (CT) and a semiautomated algorithm to calculate tumor volume and other parameter values. MATERIALS AND METHODS: This HIPAA-compliant study was institutional review board approved; informed patient consent was waived. CT scans of 15 measurable non-small cell lung cancers (in five men and 10 women; mean age, 64 years; range, 38-78 years) before and after gefitinib treatment were analyzed. A semiautomated three dimensional lung cancer segmentation algorithm was developed and applied to each tumor at baseline and follow-up. The computer calculated the greatest diameter (unidimensional measurement), the product of the greatest diameter and the greatest perpendicular diameter (bidimensional measurement), and the volume of each tumor. Exact McNemar tests were used to analyze differences in the percentage change calculated with different measurement techniques. RESULTS: The computer accurately segmented 14 of the 15 tumors. One paramediastinal tumor required manual separation from the mediastinum. Eleven (73%) of the 15 patients had an absolute change in tumor volume of at least 20%, compared with one (7%) and four (27%) patients who had similar changes in unscaled unidimensional (P < .01) and bidimensional (P = .04) tumor measurements, respectively. Seven (47%) patients had an absolute change in tumor volume of at least 30%. In contrast, at unscaled analysis, no patients at unidimensional measurement (P = .02) and two (13%) patients at bidimensional measurement (P = .06) had a change of at least 30%. CONCLUSION: Compared with the unidimensional and bidimensional techniques, semiautomated tumor segmentation enabled the identification of a larger number of patients with absolute changes in tumor volume of at least 20% and 30%. PMID- 17114631 TI - CT angiography of pulmonary arteries to detect pulmonary embolism: improvement of vascular enhancement with low kilovoltage settings. AB - PURPOSE: To retrospectively compare a low kilovoltage scanning protocol with a reduced radiation dose with a standard high kilovoltage, moderate-dose protocol for the depiction of central and peripheral pulmonary arteries at single-detector spiral computed tomography (CT). MATERIALS AND METHODS: This retrospective study had institutional review board approval; informed consent was waived. A 100-kVp protocol (volume CT dose index [CTDI(vol)], 3.4 mGy) was compared with a standard 140-kVp protocol (CTDI(vol), 10.4 mGy) in two groups that were each composed of 35 consecutive patients who were suspected of having pulmonary embolism (PE) and scanned with otherwise identical acquisition parameters and contrast material injection protocols. Mean main pulmonary artery enhancement and maximum enhancement in peripheral pulmonary arteries were compared. In a blinded evaluation, the percentages of segmental and subsegmental arteries that were considered analyzable for assessment of PE were determined. Overall image quality and delineation of various anatomic areas were subjectively assessed. Comparison of percentages of analyzable segmental and subsegmental arteries and subjective grading of image quality between the two different protocols were performed with the Mann-Whitney U test. RESULTS: There were 38 male and 24 female patients (mean age, 61 years; range, 17-86 years) in the final evaluation. There was a significantly higher average CT number in the main pulmonary artery (379 HU +/- 95) for the 100-kVp protocol than for the 140-kVp protocol (268 HU +/- 63, P < .001, two-sided t test). Maximum CT numbers in peripheral pulmonary arteries at the level of the aortic arch and lung bases, respectively, were 290 HU +/- 91 and 279 HU +/- 100 for 100 kVp and 185 HU +/- 65 and 144 HU +/- 63 for 140 kVp (P < .001). Mean percentage of subsegmental arteries considered analyzable per patient was higher for 100 kVp than for 140 kVp (segmental arteries, 92% vs 88%, P = .13; subsegmental arteries, 71% vs 55%, P < .001). Subjective grading of overall image quality and of the delineation of structures in the lungs, mediastinum, and upper abdomen did not significantly differ between protocols. CONCLUSION: At reduced radiation exposure, low kilovoltage scanning increases the percentage of central and peripheral pulmonary arteries that can be evaluated with CT angiography without a substantial decrease in image quality. PMID- 17114634 TI - Case 102: pituitary aplasia. PMID- 17114635 TI - Case 103: PHACE syndrome. PMID- 17114636 TI - The ring-around-the-artery sign. PMID- 17114637 TI - Routine CT venography after CT for pulmonary embolism: evidence-based radiology or hemorrhage from anticoagulation of false-positive deep venous thrombosis? PMID- 17114638 TI - Regarding the spoke wheel sign. PMID- 17114639 TI - Evaluation of bowel distention with a neutral contrast agent: some statistical concerns. PMID- 17114640 TI - Design and implementation of an application and associated services to support interdisciplinary medication reconciliation efforts at an integrated healthcare delivery network. AB - Confusion about patients' medication regimens during the hospital admission and discharge process accounts for many preventable and serious medication errors. Many organizations have begun to redesign their clinical processes to address this patient safety concern. Partners HealthCare, an integrated delivery network in Boston, Massachusetts, has answered this interdisciplinary challenge by leveraging its multiple outpatient electronic medical records (EMR) and inpatient computerized provider order entry (CPOE) systems to facilitate the process of medication reconciliation. This manuscript describes the design of a novel application and the associated services that aggregate medication data from EMR and CPOE systems so that clinicians can efficiently generate an accurate pre admission medication list. Information collected with the use of this application subsequently supports the writing of admission and discharge orders by physicians, performance of admission assessment by nurses, and reconciliation of inpatient orders by pharmacists. Results from early pilot testing suggest that this new medication reconciliation process is well accepted by clinicians and has significant potential to prevent medication errors during transitions of care. PMID- 17114643 TI - Up-regulation of DLK1 as an imprinted gene could contribute to human hepatocellular carcinoma. AB - Dysregulation of a genomic imprinting gene can contribute to carcinogenesis. Here, delta-like 1 homolog (Drosophila) (DLK1), a paternally expressed gene, was found to be significantly up-regulated in 60 (73.2%) of a total of 82 hepatocellular carcinoma (HCC) specimens using reverse transcription-polymerase chain reaction. In addition, immunohistochemistry staining was performed in another 88 HCC specimens, of which 50 (56.8%) cancerous tissues were considered as positive. The expression of DLK1 was obviously induced in HCC cells, Bel-7402 and MHCC-H, by a demethylation agent, 5-aza-2'-deoxycytidine. Furthermore, both demethylation of the DLK1 promoter (-565 to -362) and hypermethylation of the imprinting control domain in the region upstream of maternally expressed gene 3 were identified in a few HCC specimens. This implies that deregulation of genomic DNA methylation of the imprinted domain could be attributed to the up-regulation of DLK1 in HCC, although the undoubtedly complex mechanisms involved in the epigenetic event should be further investigated in HCC. Surprisingly, the expression of DLK1 in HCC was confirmed to be monoallelic specific, not biallelic, in three HCC specimens with a single nucleotide polymorphism as at T852C (rs2295660). Importantly, the exogenous DLK1 can significantly promote the cell proliferation of SMMC-7721 cells, a HCC cell line, whereas the suppression of endogenetic DLK1 through RNA interference can markedly inhibit cell growth, colony formation and tumorigenicity of HepG2, Hep3B and HuH-7 cells. These data suggest that DLK1 as an imprinted gene could be significantly up-regulated in HCC due to certain epigenetic events and contribute to the oncogenesis of this tumor. PMID- 17114644 TI - Ascofuranone suppresses PMA-mediated matrix metalloproteinase-9 gene activation through the Ras/Raf/MEK/ERK- and Ap1-dependent mechanisms. AB - The expression of matrix metalloproteinase-9 (MMP-9) has been implicated in the invasion and metastasis of cancer cells. Here, we found that an antitumor antibiotic, ascofuranone, inhibits invasion and MMP-9 induction induced by phorbol myristate acetate (PMA) in human cell lines. Ascofuranone also inhibits the protein expression and transcription of MMP-9 induced by tumor necrosis factor-alpha. The inhibition of MMP-9 induction was observed in human cancer cell lines as well as primary rat mesangial cells. Furthermore, as evidenced by MMP-9 promoter and electrophoretic mobility shift assays, ascofuranone specifically inhibited MMP-9 gene expression by blocking PMA-stimulated activation of activator protein-1 (AP-1). In addition, ascofuranone suppressed PMA-induced phosphorylation of Ras, Raf, MEK and extracellular signal-regulated kinase (ERK), upstream factors involved in AP-1activation, whereas the phosphorylation of p38 and JNK/mitogen-activated protein kinase was not affected by ascofuranone, suggesting that the primary target of ascofuranone for suppression of the AP-1 induction is present in upstream of ERK signaling pathway. These results suggest that the suppression of MMP-9 expression, at least in part, contributes to the antitumor activity of ascofuranone. PMID- 17114645 TI - Effects of novel 5-lipoxygenase inhibitors on the incidence of pulmonary adenomas in the A/J murine model when administered via nose-only inhalation. AB - The objective of this study was to determine the effects of 5-lipoxygenase (5-LO) inhibitors on the incidence of benzo(a)pyrene-induced pulmonary adenomas in female A/J mice. Two novel compounds, S-29606 and S-30621, and the Food and Drug Administration-approved Zileuton were investigated. S-29606 and S-30621 were selected from a group of similar active structures on the basis of local versus systemic 5-LO inhibitory activity. Preliminary studies found them to lack oral bioavailability, in direct contrast to Zileuton. Treatment was initiated 1 week following exposure to the carcinogen benzo(a)pyrene. Both S-29606 and S-30621 were dosed via nose-only inhalation 5 days a week, for 16 weeks, whereas Zileuton was administered orally. Dose levels for S-29606 and S-30621 were determined to be 220 and 430 microg/kg for the low- and high-dose groups, respectively, whereas the dose of Zileuton was 245 mg/kg. Both test compounds exhibited a significant reduction of pulmonary adenomas, compared with a positive control for high and low doses, P < 0.05. Additionally, a dose response for both S-29606 and S-30621 was observed when compared with placebo. Despite a dose 575 times greater than that of the novel test compounds, orally administered Zileuton did not produce a reduction in adenoma occurrence. The findings of this study offer compelling preliminary data for the use of S-29606 and S-30621 in further investigations of the treatment of pulmonary adenomas and support the use of inhalation drug delivery as an alternate to oral delivery for these compounds. PMID- 17114646 TI - Formation and persistence of DNA adducts formed by the carcinogenic air pollutant 3-nitrobenzanthrone in target and non-target organs after intratracheal instillation in rats. AB - Sprague-Dawley rats were treated by intratracheal instillation with a single dose of 0.2 mg/kg body wt of 3-nitrobenzanthrone (3-NBA), and whole blood, lungs, pancreases, kidneys, urinary bladders, hearts, small intestines and livers were removed at various times after administration. At five posttreatment times (2 days, 2, 10, 20 and 36 weeks), DNA adducts were analysed in each tissue by (32)P postlabelling to study their long-term persistence. 3-NBA-derived DNA adducts consisting of the same adduct pattern were observed in all tissues from animals killed between 2 days and 36 weeks and between 2 days and 20 weeks in blood. DNA isolated from whole blood contained the same 3-NBA-specific adduct pattern as that found in tissues. Although total adduct levels in the blood were much lower than those found in the lung, the target organ of 3-NBA tumourigenicity, they were related (20-25%, R(2) = 0.98) to the levels found in lung. In all organs, total adduct levels decreased over time to 20-30% of the initial levels till the latest time point (36 weeks) and showed a biphasic profile, with a rapid loss during the first 2 weeks followed by a much slower decline that reached a stable plateau at 20 weeks after treatment. These results show that uptake of 3-NBA by the lung induces high levels of specific DNA adducts in target and non-target organs of the rat. The correlation between DNA adducts in lung and blood suggests that persistent 3-NBA-DNA adducts in the blood may be useful biomarkers for human respiratory exposure to 3-NBA. PMID- 17114647 TI - Interleukin-4 inhibits caspase-3 by regulating several proteins in the Fas pathway during initial stages of human T helper 2 cell differentiation. AB - Interleukin-4 (IL-4) is the main cytokine that polarizes activated naive CD4+ T cells in the T helper 2 (Th2) direction. IL-4 also regulates the subsequent stages of Th2 cell-mediated diseases, such as allergies. We conducted a proteomics study to identify IL-4-induced differences during the initial stages of T helper cell differentiation. Primary CD4+ T lymphocytes were isolated from human cord blood, activated through CD3 and CD28, and cultured in the presence or absence of IL-4. Soluble proteins were separated by two-dimensional electrophoresis and visualized by staining with autoradiography, which indicated that at least 20 proteins might be regulated by IL-4. From this minimum of 20 stained proteins, altogether 35 proteins were identified using tandem mass spectrometry. Interestingly the fragmented form of GDP dissociation inhibitor expressed in lymphocytes/Rho GDP dissociation inhibitor 2 (Ly-GDI), a known target of Caspase-3, was observed to be down-regulated in IL-4-treated cells. It was shown in further studies that IL-4 decreases Caspase-3 activity and cell death in these cells. Neutralizing Fas-Fas ligand interaction led to decreased Caspase-3 activity and lowered Ly-GDI fragmentation. We further characterized the effects of IL-4 on the expression of main regulators in the Fas-mediated pathway. We demonstrated that IL-4 decreases expression of Fas receptor and increases expression of Bid, Bcl-2, and Bcl-xL. Importantly IL-4 significantly up-regulated the short form of c-FLIP, although the levels of c-FLIP long were unaltered after IL-4 induction. Taken together, our results indicate that IL-4 inhibits caspase activity during the initial stages of human Th2 cell differentiation by regulating expression of several key players in the Fas-induced pathway. PMID- 17114648 TI - Phosphorylation of B14.5a subunit from bovine heart complex I identified by titanium dioxide selective enrichment and shotgun proteomics. AB - Shotgun proteomics was used to study the steady phosphorylation state of NADH:ubiquinone oxidoreductase (complex I) subunits from bovine heart mitochondria. A total tryptic digestion of enzymatically active complex I was performed, and the resulting peptide mixture was subjected to phosphopeptide enrichment by the use of titanium dioxide (TiO2). The phosphopeptide-enriched fraction was separated and analyzed with nanoscale reverse-phase HPLC-ESI-MS/MS in single information-dependent acquisition. Hence two phosphorylated complex I subunits were detected: 42 kDa and B14.5a. Phosphorylation of 42-kDa subunit at Ser-59 has already been determined with fluorescent phosphoprotein-specific gel staining and mass spectrometry (Schilling, B., Aggeler, R., Schulenberg, B., Murray, J., Row, R. H., Capaldi, R. A., and Gibson, B. W. (2005) Mass spectrometric identification of novel phosphorylation site in subunit NDUFA10 of bovine mitochondrial complex I. FEBS Lett. 579, 2485-2490). In our work, this finding was confirmed using a non-gel-based approach. In addition, we report novel phosphorylation on B14.5a nuclear encoded subunit. We demonstrated evidence of the phosphorylation site at Ser-95 residue by collision-induced dissociation experiments on three different molecular ions of two tryptic phosphopeptides of B14.5a. PMID- 17114649 TI - Qualitative and quantitative analyses of protein phosphorylation in naive and stimulated mouse synaptosomal preparations. AB - Activity-dependent protein phosphorylation is a highly dynamic yet tightly regulated process essential for cellular signaling. Although recognized as critical for neuronal functions, the extent and stoichiometry of phosphorylation in brain cells remain undetermined. In this study, we resolved activity-dependent changes in phosphorylation stoichiometry at specific sites in distinct subcellular compartments of brain cells. Following highly sensitive phosphopeptide enrichment using immobilized metal affinity chromatography and mass spectrometry, we isolated and identified 974 unique phosphorylation sites on 499 proteins, many of which are novel. To further explore the significance of specific phosphorylation sites, we used isobaric peptide labels and determined the absolute quantity of both phosphorylated and non-phosphorylated peptides of candidate phosphoproteins and estimated phosphorylation stoichiometry. The analyses of phosphorylation dynamics using differentially stimulated synaptic terminal preparations revealed activity-dependent changes in phosphorylation stoichiometry of target proteins. Using this method, we were able to differentiate between distinct isoforms of Ca2+/calmodulin-dependent protein kinase (CaMKII) and identify a novel activity-regulated phosphorylation site on the glutamate receptor subunit GluR1. Together these data illustrate that mass spectrometry-based methods can be used to determine activity-dependent changes in phosphorylation stoichiometry on candidate phosphopeptides following large scale phosphoproteome analysis of brain tissue. PMID- 17114650 TI - Recent declines in hormone therapy utilization and breast cancer incidence: clinical and population-based evidence. PMID- 17114651 TI - Presence or the emergence of a F317L BCR-ABL mutation may be associated with resistance to dasatinib in Philadelphia chromosome-positive leukemia. PMID- 17114652 TI - Multicenter phase II study of irinotecan, cisplatin, and bevacizumab in patients with metastatic gastric or gastroesophageal junction adenocarcinoma. AB - PURPOSE: Bevacizumab improves survival in several solid tumor malignancies when combined with chemotherapy. We evaluated the efficacy and safety of the addition of bevacizumab to chemotherapy in the treatment of gastric and gastroesophageal junction (GEJ) adenocarcinoma. PATIENTS AND METHODS: Forty-seven patients with metastatic or unresectable gastric/GEJ adenocarcinoma were treated with bevacizumab 15 mg/kg on day 1, irinotecan 65 mg/m2, and cisplatin 30 mg/m2 on days 1 and 8, every 21 days. The primary end point was to demonstrate a 50% improvement in time to progression over historical values. Secondary end points included safety, response, and survival. RESULTS: Patient characteristics were as follows: median age 59 years (range, 25 to 75); Karnofsky performance status 90% (70% to 100%); male:female, 34:13; and gastric/GEJ, 24:23. With a median follow up of 12.2 months, median time to progression was 8.3 months (95% CI, 5.5 to 9.9 months). In 34 patients with measurable disease, the overall response rate was 65% (95% CI, 46% to 80%). Median survival was 12.3 months (95% CI, 11.3 to 17.2 months). We observed no increase in chemotherapy related toxicity. Possible bevacizumab-related toxicity included a 28% incidence of grade 3 hypertension, two patients with a gastric perforation and one patient with a near perforation (6%), and one patient with a myocardial infarction (2%). Grade 3 to 4 thromboembolic events occurred in 25% of patients. Although the primary tumor was unresected in 40 patients, we observed only one patient with a significant upper gastrointestinal bleed. CONCLUSION: Bevacizumab can be safely given with chemotherapy even with primary gastric and GEJ tumors in place. The response rate, time to disease progression (TTP), and overall survival are encouraging, with TTP improved over historical controls by 75%. Further development of bevacizumab in gastric and GEJ cancers is warranted. PMID- 17114653 TI - Meta-analysis of randomized controlled trials of prophylactic granulocyte colony stimulating factor and granulocyte-macrophage colony-stimulating factor after autologous and allogeneic stem cell transplantation. AB - PURPOSE: The primary objective of our meta-analysis was to determine whether prophylactic hematopoietic colony-stimulating factors (CSFs) after hematopoietic autologous and allogeneic stem-cell transplantation (SCT) reduced documented infections. Our secondary objectives were to determine whether prophylactic CSFs affected other outcomes including parenteral antibiotic therapy duration, infection-related mortality, graft-versus-host disease (GVHD), or treatment related mortality. METHODS: We included studies if there was random assignment between CSFs and placebo/no therapy and CSFs were given after SCT and before recovery of neutrophils. From 3,778 reviewed study articles, 34 were included based on predefined inclusion criteria. All analyses were conducted using a random effects model. RESULTS: CSFs reduced the risk of documented infections (relative risk [RR] 0.87; 95% CI, 0.76 to 1.00; P = .05) and duration of parenteral antibiotics (weighted mean difference, -1.39 days, 95% CI, -2.56 to 0.22; P = .02) but did not reduce infection-related mortality (RR, 0.76; 95% CI, 0.41 to 1.44; P = .4). CSFs did not increase grade 2 to 4 acute GVHD (RR, 1.03; 95% CI, 0.81 to 1.31; P = .8) or treatment-related mortality (RR, 1.00; 95% CI, 0.78 to 1.29; P = .98). CONCLUSION: CSFs were associated with a small reduction in the risk of documented infections but did not affect infection or treatment related mortality. PMID- 17114654 TI - C-reactive protein levels, variation in the C-reactive protein gene, and cancer risk: the Rotterdam Study. AB - PURPOSE: It remains unclear if inflammation itself may induce cancer, if inflammation is a result of tumor growth, or a combination of both exists. The aim of this study was to examine whether C-reactive protein (CRP) levels and CRP gene variations were associated with an altered risk of colorectal, lung, breast, or prostate cancer. PATIENTS AND METHODS: A total of 7,017 participants age > or = 55 years from the Rotterdam Study were eligible for analyses. Mean follow-up time was 10.2 years. High-sensitivity CRP measurements were performed to identify additional values of 0.2 to 1.0 mg/L compared with standard procedures. Genotypes of the CRP gene were determined with an allelic discrimination assay. RESULTS: High levels (> 3 mg/L) of CRP were associated with an increased risk of incident cancer (hazard ratio, 1.4; 95% CI, 1.1 to 1.7) compared with persons with low levels (< 1 mg/L), even after a potential latent period of 5 years was introduced. Although CRP seems to affect several cancer sites, the association was strongest for lung cancer (hazard ratio, 2.8; 95% CI, 1.6 to 4.9). A CRP single nucleotide polymorphism associated with decreased CRP levels was associated with an increased lung cancer risk of 2.6 (95% CI, 1.6 to 4.4) in homozygous carriers. CONCLUSION: Baseline CRP levels seem to be a biomarker of chronic inflammation preceding lung cancer, even after subtracting a 5-year latent period. Furthermore, CRP gene variation associated with low CRP blood levels was relatively common in patients with lung cancer. Both chronic inflammation and impaired defense mechanisms resulting in chronic inflammation might explain these results. PMID- 17114655 TI - Identification of tumor-specific molecular signatures in intracranial ependymoma and association with clinical characteristics. AB - PURPOSE: To delineate clinically relevant molecular signatures of intracranial ependymoma. MATERIALS AND METHODS: We analyzed 24 primary intracranial ependymomas. For genomic profiling, microarray-based comparative genomic hybridization (CGH) was used and results were validated by fluorescent in situ hybridization and loss of heterozygosity mapping. We performed gene expression profiling using microarrays, real-time quantitative reverse transcriptase polymerase chain reaction, and methylation analysis of selected genes. We applied class comparison analyses to compare both genomic and expression profiling data with clinical characteristics. RESULTS: A variable number of genomic imbalances were detected by array CGH, revealing multiple regions of recurrent gain (including 2q23, 7p21, 12p, 13q21.1, and 20p12) and loss (including 5q31, 6q26, 7q36, 15q21.1, 16q24, 17p13.3, 19p13.2, and 22q13.3). An ependymoma-specific gene expression signature was characterized by the concurrent abnormal expression of developmental and differentiation pathways, including NOTCH and sonic hedgehog signaling. We identified specific differentially imbalanced genomic clones and gene expression signatures significantly associated with tumor location, patient age at disease onset, and retrospective risk for relapse. Integrated genomic and expression profiling allowed us to identify genes of which the expression is deregulated in intracranial ependymoma, such as overexpression of the putative proto-oncogene YAP1 (located at 11q22) and downregulation of the SULT4A1 gene (at 22q13.3). CONCLUSION: The present exploratory molecular profiling study allowed us to refine previously reported intervals of genomic imbalance, to identify novel restricted regions of gain and loss, and to identify molecular signatures correlating with various clinical variables. Validation of these results on independent data sets represents the next step before translation into the clinical setting. PMID- 17114656 TI - Phase II study of consolidation paclitaxel after concurrent chemoradiation in poor-risk stage III non-small-cell lung cancer: SWOG S9712. AB - PURPOSE: A previous Southwest Oncology Group (SWOG) study (S9429) demonstrated efficacy and tolerability of concurrent chemoradiotherapy in poor-risk stage III non-small-cell lung cancer (NSCLC). This study evaluated adding consolidation paclitaxel after chemoradiotherapy for a similar patient cohort. PATIENTS AND METHODS: Patients with histologically/cytologically determined stage III NSCLC were eligible based on performance status (PS) 2 and either low albumin or weight loss more than 10%, poor pulmonary function, or comorbidities precluding cisplatin use. Treatment was carboplatin 200 mg/m2 days 1, 3, 29, and 31, and etoposide 50 mg/m2 days 1 through 4, and 29 to 32. Beginning day 1, thoracic radiation was delivered at 1.8 Gy in 25 fractions plus 16-Gy boost (total dose, 61 Gy). Patients without disease progression received paclitaxel 175 mg/m2 every 21 days for three cycles. RESULTS: Characteristics of 87 eligible patients were age 51 to 82 years; 57% PS 0 to 1, 43% PS 2; and 51% stage IIIA, 49% stage IIIB. Toxicities of concurrent chemoradiotherapy included grade 3 esophagitis (7%) and grade 3/4 neutropenia (43%). Fifty-four assessable patients received paclitaxel consolidation. Four treatment-related deaths occurred during chemoradiotherapy and four occurred during consolidation. Overall response rate was 53%. Median progression free- and overall survival were 6.1 and 10.2 months, respectively. One- and 2-year survival rates were 43% and 25%. CONCLUSION: Compared with a previous SWOG trial in a similar patient population, the addition of consolidation paclitaxel after chemoradiotherapy resulted in increased toxicity without a survival advantage. More PS 2 patients (43% v 18%) enrolled onto S9712, which may explain increased toxicity and lack of benefit. The optimal chemoradiotherapy approach for poor-risk patients remains to be defined. PMID- 17114657 TI - Phase II study of etoposide and cisplatin with concurrent twice-daily thoracic radiotherapy followed by irinotecan and cisplatin in patients with limited disease small-cell lung cancer: West Japan Thoracic Oncology Group 9902. AB - PURPOSE: We initially conducted a randomized phase II study to compare irinotecan and cisplatin (IP) versus irinotecan, cisplatin, and etoposide (IPE) after etoposide and cisplatin (EP) with concurrent twice-daily thoracic radiotherapy (TRT) in limited-disease small-cell lung cancer (LD-SCLC). We amended the protocol to evaluate IP after EP with concurrent twice-daily TRT in a single-arm phase II study because of an unacceptable toxicity in IPE. PATIENTS AND METHODS: Previously untreated patients with LD-SCLC were treated intravenously with etoposide 100 mg/m2 on days 1 through 3 and cisplatin 80 mg/m2 on day 1 with concurrent twice-daily TRT (1.5 Gy per fraction, a total dose of 45 Gy) beginning on day 2 followed by three cycles of irinotecan 60 mg/m2 on days 1, 8, and 15 and cisplatin 60 mg/m2 on day 1 of a 4-week cycle. RESULTS: Of the 51 patients enrolled, 49 patients were assessable for response and toxicity. The overall response rate and complete response rate were 88% and 41%, respectively. The median survival time for all patients was 23 months. The 2-year and 3-year survival rates were 49% and 29.7%, respectively. The median progression-free survival was 11.8 months. The major toxicities observed were neutropenia (grade 4, 84%), febrile neutropenia (grade 3, 31%), infection (grade 3 to 4, 33%), electrolytes imbalance (grade 3 to 4, 20%), and diarrhea (grade 3 to 4, 14%). CONCLUSION: EP with concurrent twice-daily TRT followed by the consolidation of IP appears to be an active regimen which deserves further phase III testing in patients with LD-SCLC. PMID- 17114658 TI - Phase II trial of cetuximab in patients with previously treated non-small-cell lung cancer. AB - PURPOSE: To determine the efficacy of cetuximab in patients with recurrent or progressive non-small-cell lung cancer (NSCLC) after receiving at least one prior chemotherapy regimen. PATIENTS AND METHODS: This was an open-label, phase II study of patients with epidermal growth factor receptor (EGFR) -positive and EGFR negative advanced NSCLC with Eastern Cooperative Oncology Group performance status 0 to 1. Patients received cetuximab 400 mg/m2 intravenously (IV) during 120 minutes on week 1 followed by weekly doses of cetuximab 250 mg/m2 IV during 60 minutes. A cycle was considered as 4 weeks of treatment and therapy was continued until disease progression or intolerable toxicities. The primary end point was to assess response rate. Secondary end points included an estimation of time to progression and survival. RESULTS: Patient and disease characteristics (n = 66) included EGFR-positive status (n = 60); EGFR-negative status (n = 6); number of prior regimens (one, n = 28; two, n = 27; > or = three, n = 11); male (n = 41); female (n = 25); adenocarcinoma (n = 36); and smoking status (never, n = 13; former, n = 45; current, n = 8). Grade 3/4 toxicities included acne-like rash (6.1%), anaphylactic reactions (1.5%), and diarrhea (1.5%). The response rate for all patients (n = 66) was 4.5% (95% CI, 0.9% to 12.7%) and the stable disease rate was 30.3% (95% CI, 19.6% to 42.9%). The response rate for patients with EGFR-positive tumors (n = 60) was 5% (95% CI, 1.0% to 13.9%). The median time to progression for all patients was 2.3 months (95% CI, 2.1 to 2.6 months) and median survival time was 8.9 months (95% CI, 6.2 to 12.6 months). CONCLUSION: Although the response rate with single-agent cetuximab in this heavily pretreated patient population with advanced NSCLC was only 4.5%, the disease control rates and overall survival seem comparable to that of pemetrexed, docetaxel, and erlotinib in similar groups of patients. PMID- 17114659 TI - Phase II study of temozolomide in relapsed or refractory high-risk neuroblastoma: a joint Societe Francaise des Cancers de l'Enfant and United Kingdom Children Cancer Study Group-New Agents Group Study. AB - PURPOSE: To determine the response rate (RR) of neuroblastoma (NB) in children to temozolomide (TMZ), and evaluate the duration of response and tolerance of the drug in this patient population. PATIENTS AND METHODS: A multicenter, phase II evaluation of an oral, daily schedule of TMZ (200 mg/m2/d x 5 days every 28 days) was undertaken in children with refractory or relapsed high-risk NB (metastatic or localized with Myc-N amplification). Response assessment was based on imaging with two-dimentional measurement of disease and meta-iodobenzylguanidine (MIBG) score. Activity was defined by a reduction in lesion size or isotope uptake at anytime. Methodology included a two-step design using Fleming's method with a first step of 15 patients and a second of 10 additional patients if two to four responses had been observed in the first cohort. All data was centrally reviewed by a panel. RESULTS: Twenty-five assessable patients were recruited over a 14 month period in 14 centers and received 94 cycles of chemotherapy. Twenty-three patients had metastatic NB either refractory (n = 9) or in relapse (n = 14). Grade 3 or 4 thrombocytopenia was the most frequent toxicity (16% of cycles). Myelosuppression resulted in treatment delays and dose reductions (24% and 21% of cycles, respectively). Response (complete response, very good partial response, or partial response) was observed in five patients (RR = 20% +/- 8%) with a median duration of 6 months and an objective or mixed response in five additional patients. CONCLUSION: Temozolomide shows activity in heavily pretreated patients with NB, and deserves further evaluation in combination with another drug. PMID- 17114660 TI - Communication about prognosis between parents and physicians of children with cancer: parent preferences and the impact of prognostic information. AB - PURPOSE: Concerns about the harms of prognostic information, including distress and loss of hope, cause some physicians to avoid frank disclosure. We aimed to determine parent preferences for prognostic information about their children with cancer and the results of receiving such information. PATIENTS AND METHODS: We surveyed 194 parents of children with cancer (overall response rate, 70%), treated at the Dana-Farber Cancer Institute and Children's Hospital (Boston, MA) and the children's physicians. Our main outcome measure was parent rating of prognostic information as extremely or very upsetting. RESULTS: The majority of parents desired as much information about prognosis as possible (87%) and wanted it expressed numerically (85%). Although 36% of parents found information about prognosis to be extremely or very upsetting, those parents were more likely to want additional information about prognosis than those who were less upset (P = .01). Parents who found information upsetting were no less likely to say that knowing prognosis was important (P = .39), that knowing prognosis helped in decision making (P = .40), or that hope for a cure kept them going (P = .72). CONCLUSION: Although many parents find prognostic information about their children with cancer upsetting, parents who are upset by prognostic information are no less likely to want it. The upsetting nature of prognostic information does not diminish parents' desire for such information, its importance to decision making, or parents' sense of hope. PMID- 17114661 TI - Irinotecan plus temozolomide for relapsed or refractory neuroblastoma. AB - PURPOSE: To report on an irinotecan and temozolomide regimen for neuroblastoma (NB). Quality of life and minimizing toxicity were major considerations. PATIENTS AND METHODS: The plan stipulated 5-day courses of irinotecan 50 mg/m2 (1-hour infusion) and temozolomide 150 mg/m2 (oral) every 3 to 4 weeks, with a pretreatment platelet count more than 30,000/microL. Granulocyte colony stimulating factor was used when the absolute neutrophil count was less than 1,000/microL. RESULTS: Forty-nine NB patients received 1 to 15 courses (median, 5). Gastrointestinal and myelosuppressive toxicities were readily managed. Lymphocyte responses to phytohemagglutinin after 2 to 10 courses (median, 3.5) were normal in 10 of 10 patients treated after nonimmunosuppressive therapy, and normalized in five of seven patients first treated less than 2 months after high dose alkylators. Of 19 patients treated for refractory NB and assessable for response, nine showed evidence of disease regression, including two complete responses and seven objective responses. Of 17 patients treated for progressive disease, three showed evidence of disease regression, including one partial response and two objective responses. Multiple courses entailed no cumulative toxicity and controlled disease for prolonged periods in many patients, including some who were unable to complete prior treatments because of hematologic, infectious, cardiac, or renal problems. CONCLUSION: This regimen has anti-NB activity, spares vital organs, is feasible with poor bone marrow reserve, causes limited immunosuppression, and allows good quality of life. PMID- 17114662 TI - Changes in attentional performance of children and young adults with localized primary brain tumors after conformal radiation therapy. AB - PURPOSE: To prospectively assess the impact of conformal radiation therapy (CRT) and demographic and clinical variables on four measures of attention in pediatric and young adult patients with localized primary brain tumors. PATIENTS AND METHODS: We prospectively evaluated 120 patients with primary brain tumors, ages 2 to 24.4 years (median, 9.2 years). Evaluations were done using the computerized Conners' Continuous Performance Test (CCPT). We analyzed errors of omission (inattentiveness), errors of commission (impulsivity), reaction time, and an overall index of performance before CRT, weekly during CRT, and serially up to 60 months after the start of CRT. RESULTS: Before CRT, patients exhibited mild inattentiveness. During CRT, impulsivity decreased significantly (P = .002). After CRT, inattentiveness increased significantly (P = .03), and global attention disorders were associated with craniopharyngioma (P < .0001), supratentorial tumors (P = .008), optic pathway and diencephalic tumors (P = .012), and subtotal resection of the tumor (P = .010). CONCLUSION: Brain tumors and their treatment impair sustained attention and reaction time. A decline in impulsivity and relative stability of the other CCPT scores over the course of CRT demonstrated the absence of early radiation-related cognitive sequelae. Local tumor effects, initial surgical intervention, and focal irradiation of central structures contribute to long-lasting attentional problems in pediatric and young adult patients. PMID- 17114663 TI - Plasma osteopontin is an independent prognostic marker for head and neck cancers. AB - PURPOSE: To confirm the relationship between plasma osteopontin (OPN) levels and treatment outcomes in head and neck squamous cell carcinoma (HNSCC) patients in an expanded study. PATIENTS AND METHODS: One hundred forty patients with newly diagnosed HNSCC were enrolled onto this study, 54 previously reported and 86 new patients. Pretreatment plasma OPN levels were assessed in all patients by an enzyme-linked immunosorbent assay method. OPN levels were correlated to treatment outcomes in the new group of patients. Detailed analyses were also performed on the relationship between OPN and tumor control rate, event-free survival (EFS), and postrelapse survival for the entire group. RESULTS: Using a previously defined cut off point of 450 ng/mL, there was a significant correlation between OPN and freedom-from-relapse (P = .047), overall survival (P = .019), and EFS (P = .023) in the new, independent patient cohort (n = 86). Sequence of event analyses using the entire group (N = 140) revealed that OPN was an independent prognostic factor for initial tumor control, EFS in those who have achieved tumor control, and postrelapse survival. CONCLUSION: In this expanded study, we were able to replicate the prognostic significance of OPN using a predefined cut off point in an independent patient group and demonstrated that plasma OPN is an independent prognostic marker for HNSCC. PMID- 17114664 TI - Patient and treatment factors associated with complications after prostate brachytherapy. AB - PURPOSE: To assess the prevalence and predictors of complications after prostate brachytherapy in a population-based sample of older men. PATIENTS AND METHODS: We analyzed claims for Medicare-enrolled men older than age 65 years living in Surveillance, Epidemiology, and End Results (SEER) surveillance areas diagnosed with prostate cancer from 1991 to 1999 who underwent brachytherapy as initial treatment. RESULTS: There were 5,621 men who had brachytherapy with at least 2 years of follow-up. A complication diagnosis or invasive procedure occurred in 54.5% of men within 2 years, with 14.1% undergoing an invasive procedure. Urinary, bowel, and erectile morbidity rates were 33.8%, 21.0%, and 16.7%, respectively, and invasive procedure rates were 10.3%, 0.8%, and 4.0%, respectively. On multivariable analysis, combined urinary diagnoses and invasive procedures (obstruction, incontinence, bleeding, fistula) were associated with older age (P < .01), nonwhite race (odds ratio [OR], 1.30; P = .01), low income (OR, 1.74; P < .01), external-beam radiotherapy (EBRT; OR, 0.85; P = .01), androgen deprivation (OR, 1.31; P < .01), later year of brachytherapy (OR, 1.03/yr; P = .02), higher Charlson comorbidity score (P < .01), and prior transurethral resection of the prostate (OR, 1.65; P < .01). Bowel morbidity (bleeding/proctitis, injury) was associated with older age (P = .04), EBRT (OR, 1.46; P < .01), later year (OR, 1.04/yr; P < .01), higher Charlson score (P = .01), and inflammatory bowel disease (OR, 2.60; P < .01). Erectile morbidity was associated with younger age (P < .01), nonwhite race (OR, 1.37; P < .01), AD (OR, 1.18; P = .04), and later year (OR, 1.08/yr; P < .01). Invasive procedure rates declined with later year of brachytherapy (OR, 0.93/yr; P < .01). CONCLUSION: Morbidity after prostate brachytherapy was common, though invasive procedures were required infrequently. Invasive procedures for complications declined during the 1990s, suggesting technical improvement with experience. PMID- 17114665 TI - Aromatase inhibitors and bone health in women with breast cancer. PMID- 17114666 TI - Thalidomide use and digital gangrene. PMID- 17114667 TI - Mesothelioma of the tunica vaginalis complicated by chyluria. PMID- 17114668 TI - Pneumocystis carinii pneumonia during dose-dense chemotherapy for breast cancer. PMID- 17114669 TI - Successful treatment of syncope in head and neck cancer with induction chemotherapy. PMID- 17114670 TI - The ideal oncology curriculum for medical students. PMID- 17114671 TI - American Society of Clinical Oncology recommendations on fertility preservation should be implemented regardless of disease status or previous treatments. PMID- 17114672 TI - Fertility preservation in (breast) cancer patients: is it safe? PMID- 17114673 TI - In vitro maturation is an efficient technique to generate oocytes and should be considered in combination with cryopreservation of ovarian tissue for preservation of fertility in women. PMID- 17114674 TI - Sensitivity of remediastinoscopy: influence of adhesions, multilevel N2 involvement, or surgical technique? PMID- 17114675 TI - A century of rickettsiology: emerging, reemerging rickettsioses, clinical, epidemiologic, and molecular diagnostic aspects and emerging veterinary rickettsioses: an overview. AB - This overview summarize the salient features of advances in the epidemiology, vectors, and clinical and laboratory diagnoses of rickettsiology. Presentations on veterinary rickettsiology highlight the importance of the rickettsiae in animal husbandry. PMID- 17114676 TI - Insights into mechanisms of bacterial antigenic variation derived from the complete genome sequence of Anaplasma marginale. AB - Persistence of Anaplasma spp. in the animal reservoir host is required for efficient tick-borne transmission of these pathogens to animals and humans. Using A. marginale infection of its natural reservoir host as a model, persistent infection has been shown to reflect sequential cycles in which antigenic variants emerge, replicate, and are controlled by the immune system. Variation in the immunodominant outer-membrane protein MSP2 is generated by a process of gene conversion, in which unique hypervariable region sequences (HVRs) located in pseudogenes are recombined into a single operon-linked msp2 expression site. Although organisms expressing whole HVRs derived from pseudogenes emerge early in infection, long-term persistent infection is dependent on the generation of complex mosaics in which segments from different HVRs recombine into the expression site. The resulting combinatorial diversity generates the number of variants both predicted and shown to emerge during persistence. PMID- 17114677 TI - Rickettsiosis in Europe. AB - In Europe, rickettsioses are long-known infectious diseases. Until recently, it was thought that Mediterranean spotted fever due to Rickettsia conorii was the only tick-borne rickettsiosis in Europe. In the last decade new Rickettsia spp. have been implicated in human pathology (R. slovaca, R. sibirica mongolotimonae, R. helvetica). Furthermore, cases of infection due to flea-borne rickettsioses (R. typhi, R. felis) have been described. Finally, although no outbreak of epidemic typhus has been reported yet in central and southern Europe, we should be aware of the possibility of reemergence of this disease in Europe. Other rickettsioses exist that have not yet been implicated in human pathology. We should consider that climate changes and other factors could contribute to the emergence and reemergence of other new diseases. PMID- 17114678 TI - Epidemiology of rickettsioses in North Africa. AB - The first description of Mediterranean spotted fever (MSF) was made by Conor and Brush in 1910 in Tunisia, where, at the same time, Nicolle described the role of lice in transmission of epidemic typhus. However, along this century, there have been few and fragmentary reports about ecology and epidemiology of rickettsioses in North Africa. This region was always considered, for these diseases, like other Mediterranean regions. The most human tick-borne rickettsiosis known to occur in North Africa is MSF caused by R. conorii and transmitted by the brown dog tick, Rhipicephalus sanguineus. Recent studies showed that other arthropode transmitted rickettsiae are prevalent in North Africa: R. aeschlimannii, R. massiliae, and R. felis. Moreover, R. felis and R. aeschlimannii human infection were respectively confirmed, by serology in Tunisia, and by PCR in Morocco. The seroprevalence of R. conorii among healthy population was ranging from 5% to 8% in most of the countries. Epidemiological and clinical features are frequently resumed in an eruptive fever with eschar occurring in hot season in rural areas. Typhus group rickettsioses, caused by R. typhi and R. prowazekii are less frequently reported than in the 1970s. Seroprevalence of R. typhi among blood donors was from 0.5% to 4%. In Algeria about 2% of febrile patients had R. prowazekii antibodies. Moreover, reemerging threat of epidemic typhus should be considered, after the two cases recently diagnosed in the highlands of Algeria. Murine typhus, considered as "benign" typhus, is underestimated. When R. typhi was inserted in serologic tests, murine typhus became more frequently confirmed. In a recent study in Central Tunisia, we confirmed an emergence of murine typhus mistaken for R. conorii or viral infection. In addition to typhus surveillance, future studies have to determine which spotted fever group rickettsiae are prevalent in vectors and in human pathology. PMID- 17114679 TI - Rickettsioses in sub-Saharan Africa. AB - Although rickettsioses are among the oldest known vector-borne zoonoses, several species or subspecies of rickettsias have been identified in recent years as emerging pathogens throughout the world including in sub-Saharan Africa. To date, six tick-borne spotted fever group pathogenic rickettsias are known to occur in sub-Saharan Africa, including Rickettsia conorii conorii, the agent of Mediterranean spotted fever; R. conorii caspia, the agent of Astrakhan fever; R. africae, the agent of African tick-bite fever; R. aeschlimannii; R. sibirica mongolitimonae; and R. massiliae. On the other hand, fleas have long been known as vectors of the ubiquitous murine typhus, a typhus group rickettsiosis induced by R. typhi. However, a new spotted fever rickettsia, R. felis, has also been found to be associated with fleas, to be a human pathogen, and to be present in sub-Saharan Africa. Finally, R. prowazekii the agent of louse-borne epidemic typhus continues to strikes tens to hundreds of thousands of persons who live in Sub-Saharan with civil war, famine and poor conditions. We present an overview of these rickettsioses occurring in sub-Saharan Africa, focusing on the epidemiological aspects of emerging diseases. PMID- 17114680 TI - Rickettsial diseases in Russia. AB - Currently, several rickettsioses are officially being reported in the Russian Federation. These are epidemic typhus and Brill-Zinsser disease, both caused by Rickettsia prowazekii which has a historic prevalence in Russia. Nowadays only single sporadic cases of R. prowazekii infection are reported. The last significant outbreak occurred in 1997 in a mental nursing home, where 29 cases were identified. Registered morbidity of typhus in Russia varies from 0 to 0.01 per thousand for the last decade. Siberian tick typhus, caused by R. sibirica, is registered on a large territory from Pacific coasts to Western Siberia, and its incidence continuously increases, varying between 2.5 and 4.0 thousand officially registered cases per year. Astrakhan spotted fever, caused by R. conorii subsp. caspia has been recognized since 1983. Recently, Far Eastern tick-borne rickettsiosis, caused by R. heilongjiangensis, has been described. Several other pathogenic spotted fever group rickettsiae have been detected and isolated from ticks in Russia; however, they have not yet been linked with clinical cases in these regions. PMID- 17114681 TI - Rickettsioses in Japan and the far East. AB - Three rickettsial diseases are known to exist in Japan currently: Japanese spotted fever (JSF), Tsutsugamushi disease (TD; scrub typhus), and Q fever. Since April 1999, the system for infection control and prevention in Japan has changed drastically. JSF, Q fever, and TD, as emerging infectious diseases, are designated as national notifiable diseases.The geographic distribution of JSF patients is along the coast of central and southwestern Japan, whereas TD and Q fever occur almost all over the country. The number of JSF patients reported was 216 cases during 1984-1998 and 268 cases, under the revised law, in 1999-2004. About 300-1000 cases of TD occur every year, and 7-46 cases of Q fever in 1999 2004. The number of cases of JSF and its endemic area are gradually increasing. There was only one fatality due to JSF until 2003, whereas two patients died of JSF in 2004, so JSF is still a life-threatening disease in Japan. Treatment of fulminant JSF consists of prompt administration of a combination of tetracycline and quinolone. Recent tick surveys revealed that the most probable vectors of JSF are Haemophysalis flava and Haemophysalis hystericis. In addition to R. japonica, two serotypes or species of spotted fever group rickettsiae have been isolated from ticks in Japan; one is closely related to R. helvetica and the other is a new genotype of unknown genotype AT, which is closely related to a Slovakian genotype. These serotypes are of uncertain clinical significance. Epidemiology of rickettsioses in the Far East is mentioned briefly. PMID- 17114682 TI - Rickettsioses in Australia. AB - Australia, an island continent in the southern hemisphere, has a range of rickettsial diseases that include typhus group rickettsiae (Rickettsia typhi), spotted fever group rickettsiae (R. australis, R. honei), scrub typhus group rickettsiae (R. tsutsugamushi), and Q fever (C. burnetii). Our knowledge of Australian rickettsiae is expanding with the recognition of an expanded range of R. honei (Flinders Island spotted fever) to Tasmania and southeastern mainland Australia (not just on Flinders Island), and the detection of a new SFG species (or subspecies), tentatively named "R. marmionii" in the eastern half of Australia. This rickettsia causes both acute disease (7 cases, recognized so far) and is also associated (as a "R. marmionii" bacteriaemia) with patients having a chronic illness. The significance of the latter is under investigation. It may be a marker of autoimmune disease or chronic fatigue in some patients. PMID- 17114683 TI - Far eastern tick-borne rickettsiosis: identification of two new cases and tick vector. AB - We recently reported the first documented cases of a new rickettsial disease caused by Rickettsia heilongjiangensis in the Russian Far East (Far Eastern tick borne rickettsiosis). Here we report the amplification of DNA of R. heilongjiangensis from both the skin biopsy of an acutely ill patient and the tick removed from him prior to the disease development. The tick has been identified as Haemaphysalis spp. The clinical picture was that of a spotted fever group rickettsiosis and a seroconversion was noted with R. heilongjiangensis antigen. Screening testing of both species of Haemaphysalis ticks inhabiting Russian Far Eastern regions showed that up to 28.13% of H. concinnae and 4.48% of H. japonica douglasii ticks harbor R. heilongjiangensis. It has been concluded that H. concinnae may serve as the main vector for the transmission of R. heilongjiangensis. H. japonica douglasii ticks harbor several varieties of rickettsiae. DNA of "Candidatus Rickettsia tarasevichiae," previously found in Ixodes persulcatus ticks, was amplified from one male tick. Two sequenced complete gltA genes belong to the novel spotted fever group rickettsial species provisionally called here "Candidatus Rickettsia principis" variants Hjd54 and Hjd61. The rate of infection has been found to be not higher than 1.5%. PMID- 17114684 TI - Seroprevalence of Anaplasma phagocytophilum among forestry rangers in northern and northeastern Poland. AB - The aim of this study was to evaluate the seroprevalence of A. phagocytophilum in a group of workers from 13 forest management areas of northern and northeastern Poland. A total of 478 sera were tested by indirect immunofluorescence assay (IFA). Elevated IgG antibody titers were detected in 46 samples (9.6%). Of these, 34 (73.9%) had a titer of 1:64, 7 (15.2%) 1:128, and 5 (10.9%) 1:256. All seropositive persons disclaimed any clinical symptoms of HGA. The antibodies prevalent in persons with extreme outdoor activity and in indoor workers was comparable-9.6 % and 9.7%, respectively. Moreover, participants had no statisfically significant differences in the prevalence of antibodies related to sex, age, or years of employment. PMID- 17114685 TI - Seroprevalence of human anaplasmosis in slovene forestry workers. AB - The aim of the present study was to establish the prevalence and incidence of symptomatic and asymptomatic infection with Anaplasma phagocytophilum during the period of tick activity and to compare the risk of infection for forestry workers and indoor workers in Slovenia. PMID- 17114686 TI - Molecular epidemiology of human and bovine anaplasmosis in southern Europe. AB - The genus Anaplasma (Rickettsiales: Anaplasmataceae) includes several pathogens such as A. marginale and A. phagocytophilum that have an impact on veterinary and human health. In this study, we characterized A. marginale and A. phagocytophilum infections in humans, wild and domestic animals, and ticks in southern Europe (particularly in south-central Spain and in Sicily) by means of serologic study, PCR, and sequence analysis of major surface proteins (msp) 1alpha and 4 and 16S rDNA. The results suggest that A. marginale infections in this region are maintained in cattle and deer, with ticks and tabanids serving as biological and mechanical vectors of the pathogen, respectively. Infections with A. phagocytophilum may occur in humans and are maintained in cattle, donkeys, deer, and birds and are most likely transmitted by several tick species with as yet an unknown role as reservoir hosts for other wild and domesticated mammals. The presence of concurrent infections in cattle and deer suggests that these pathogens may multiply in the same reservoir host and illustrates the complexity of the epidemiology of bovine and human anaplasmosis in this region. PMID- 17114687 TI - Human exposure to Anaplasma phagocytophilum in Portugal. AB - A retrospective study to detect Anaplasma phagocytophilum antibodies by indirect immunofluorescence (IFA) and Western blot (WB) assay was conducted in 367 potentially exposed patients from Portugal. The study included 26 patients with confirmed Lyme borreliosis (LB), 77 with suspected LB, 264 seronegative patients studied for possible tick-transmitted LB and boutonneuse fever (LB/BF) infection, and 96 healthy blood donors. Overall, patients with LB and suspected LB (n = 2 [7.7%] and n = 6 [7.8%], respectively) were more often seropositive (n = 8 [7.8%]; P < 0.001), whereas only 1 (0.4%; P = 0.046) patient in the LB/BF seronegative group had confirmed disease. This study is the first evidence of human exposure to A. phagocytophilum or an antigenically similar bacterium in Portugal, and suggests that LB patients are significantly more likely to contact A. phagocytophilum. PMID- 17114688 TI - Human granulocytic anaplasmosis in northeastern Italy. AB - Sporadic cases of human granulocytic anaplasmosis (HGA) have been reported in areas with a high prevalence of tick-borne diseases (TBDs) in Europe. We aimed at estimating the sero-prevalence of A. phagocytophilum and other TBDs in northeastern Italy in outpatients with a history of recent tick bite or suspected TBD. In the 1-year study, 79 patients were enrolled and 30 (38%) received a diagnosis of TBD: 24 (30%) with Lyme disease and 5 (6%) with HGE. Our findings indicate the presence of HGA in northeastern Italy; so, since co-infection with Lyme disease appeared to be frequent, physicians assessing patients after a tick bite should consider HGA in the diagnosis. PMID- 17114689 TI - Human infection with Ehrlichia canis accompanied by clinical signs in Venezuela. AB - A total of 20 human patients with clinical signs compatible with human monocytic ehrlichiosis (HME), who were admitted to the emergency clinic in Lara State, Venezuela, were studied. Thirty percent (6/20) patients were positive for Ehrlichia canis 16S rRNA on gene-specific polymerase chain reaction (PCR). Compared with the U.S. strains, 16S rRNA gene sequences from all six patients had the same base mutation as the sequence of the E. canis Venezuelan human Ehrlichia (VHE) strain previously isolated from an asymptomatic human. This study is the first report of E. canis infection of human patients with clinical signs of HME. PMID- 17114690 TI - Human monocytic ehrlichiosis and human granulocytic anaplasmosis in the United States, 2001-2002. AB - The epidemiologic features are described of cases of human monocytic ehrlichiosis and human granulocytic anaplasmosis in the United States. PMID- 17114691 TI - Anthropogenic effects on changing Q fever epidemiology in Russia. AB - In the northwestern region of Russia (Leningrad province) cattle is proved to be the main source of C. burnetii infection in humans, both in menaced professionals and in formally nonmenaced groups. Liquidation of specialized cattle-breeding complexes (with their well-organized veterinary surveillance) and broadening of the circle of non-professionals that contact with agriculture or domestic animals infected with C. burnetii provide the prerequisites to Q fever spreading among various groups of population. PMID- 17114692 TI - Human Coxiella burnetii infections in regions of Bosnia and Herzegovina, 2002. AB - Acute infections in humans and animals caused by Coxiella burnetii (C. burnetii) are becoming an important medical problem for Bosnia and Herzegovina (B&H). From a clinical and epidemiological aspect, Q fever represents a complex medical problem, considering that one of the highest incidence rates of Q fever in Europe has been recorded during the last few years in B&H. The first case of this disease in B&H was described in 1950, by Muray et al., and the first epidemic, with 16 infected individuals, was recorded the same year. Confirmed animal infections by C. burnetii in B&H were first reported in 1985 when, of all tested sheep, positive results were found in 12.4%. During 2001, 2.11% of tested sheep and goats were found to have a positive result, which was also confirmed by studies from the following years in particular regions of B&H. These studies suggest that endemic loci of infected animals are established in particular geographic regions in B&H, which is important to emphasize for better understanding of the sources and routes of C. burnetii transmission to the human population. This conclusion is based on the studies from 2000, when 2.17% of positive cattle, 1.85% of positive sheep, and 0.27% of positive goats were registered. During the same period, in B&H, in 6 different regions, 156 individuals with Q fever were registered as were 3 separate epidemics with 115 infected individuals. Official data on the number of detected animal C. burnetii infections during 2002 suggest that 10 positive cattle and 88 positive sheep or goats were registered. During 2003, 24 positive cattle, 29 positive goats, and 167 positive sheep were detected, while in 2004, 71 positive cattle, 4 positive goats, 37 positive sheep, and 72 positive animals from the sheep-goat group were registered. According to official reports from 2001, 19 individuals with Q fever were registered in B&H, while in 2002, the number of infected individuals increased to 250. In five cantons in B&H, 43 infected individuals were registered during 2002, while in Republika Srpska of B&H, 207 infected individuals in the region of Banja Luka were registered. From 1998 to 2003, 373 individuals with Q fever were reported in B&H, whereof 265 individuals (71.04%) were infected during epidemics, and 108 (28.95%) sporadically. Q fever incidence rates in B&H were high during 1998 (5.68%ooo) and very high in 2000, with 115 individuals with an acute clinical form and an incidence rate of 6.95%ooo. The incubation time varied between 9 and 28 days. PMID- 17114693 TI - Gipuzkoa, Basque Country, Spain (1984-2004): a hyperendemic area of Q fever. AB - Overall 1,261 cases of Q fever were diagnosed between 1984 and 2004 in Gipuzkoa (Basque Country, Spain). Most (75.5%) of the cases ocurred in subjects 15-45 years of age. A total of 79.5% of the cases (n = 1003) ocurred between January and June. The annual incidence for acute Q fever in Gipuzkoa was 7.7, 15.8, 9.6, and 5.7 for the periods 1984-1989, 1990-1994, 1995-1999, and 2000-2004, respectively. In 94% of the cases IgM titer was >/=1/256. The most frequent clinical manifestation was pneumonia (79%). Only two cases of chronic Q fever were detected during the 21 years studied. PMID- 17114694 TI - Serotesting of human Q fever distribution in Bosnia and Herzegovina. AB - Q fever is a zoonotic disease with worldwide distribution. It occurs in different geographic regions and climate zones. From 1990 till the end of 1997, only three infected individuals were registered in Bosnia and Herzegovina, during the year 1991, with the incidence of 0.05% 000. From 1996 onward, there was a sudden aggravation of epizoological and epidemiological situation in particular regions of Bosnia and Herzegovina. We performed serotesting during the 4-year period from 2000 to 2003. We tested serum samples from 708 individuals from different regions of Bosnia and Herzegovina. Q fever was serologically diagnosed in 249 individuals. The overall seroprevalence was 35.2%. The acute disease form was confirmed in 79.9% of the whole seropositive sample. Most of the Q seropositive individuals were from Kakanj (17.3%), Mostar (15.3%), Sarajevo (12.5%), Bihac (9.6%), Zenica (9.2%), Gornji Vakuf (8.9%), Tesanj (4.4%), Visoko (2.8%), and Travnik (2.4%). The number and distribution of seropositive individuals suggests that Q fever is endemic in Bosnia and Herzegovina. PMID- 17114695 TI - Ticks and tick-borne Rickettsiae surveillance in Montesinho Natural Park, Portugal. AB - This study constitutes the first contribution to the knowledge of tick dynamics and its implication in the epidemiology of rickettsial diseases in Montesinho Natural Park (MNP), Braganca district of Portugal. Of 76 ticks collected, 12 (15.8%) were Dermacentor (D.) marginatus, 36 (47.4%) D. reticulatus, and 28 (36.8%) Rhipicephalus (R.) sanguineus. Isolation assays were performed by shell vial technique on 41 ticks. Israeli spotted fever strain was an isolate from R. sanguineus, and three isolates of Rickettsia slovaca were obtained from D. reticulatus. All 76 ticks were screened by PCR for Rickettsia sp., Ehrlichia (E.) chaffeensis, and Anaplasma (A.) phagocytophilum. Rickettsia RpA4 strain DNA was detected in 10 D. marginatus and 2 D. reticulatus, and Israeli spotted fever strain in 1 R. sanguineus. No E. chaffeensis or A. phagocytophilum infection was detected. New host records are provided for D. reticulatus. Also described for the first time in Portugal is the isolation of R. slovaca from D. reticulatus and the isolation of Israeli spotted fever strain from R. sanguineus. This confirms the association of the last rickettsiae strain with the same vector tick as previously described in Israel and Sicily. PMID- 17114696 TI - Current knowledge of rickettsial diseases in Italy. AB - Rickettsial diseases continue to be the cause of serious health problems in Italy. From 1998 to 2002, 4,604 clinical cases were reported, with 33 deaths in the period from 1998 to 2001. Almost all the cases reported in Italy are cases of Mediterranean spotted fever (MSF). Other rickettsioses that have been historically documented are murine typhus and epidemic typhus. Since 1950, only sporadic cases of murine typhus have been reported, and Italy currently appears to be free of epidemic typhus. As in other European countries, imported cases of rickettsialpox, African tick-bite fever (ATBF), and scrub typhus have been reported. In 2004, three cases of a mild form of rickettsiosis were serologically attributed to Rickettsia helvetica. PMID- 17114697 TI - No serological evidence for rickettsial diseases among Danish elite orienteerers. AB - A series of sudden unexpected cardiac deaths among Swedish elite orienteerers in the 1980s have resulted from the combination of infectious diseases and physical exercise. Studies in the late 1990s have pointed to Chlamydia and Barontella, which both had a high seroprevalence among Swedish elite orienteerers. We conducted a case-control study aimed to elucidate the serologic prevalence of rickettsial diseases among Danish elite orienteerers. Ticks are known as vectors for some rickettsial diseases. None of the orienteerers had a positive antibody titer against any of the tested Rickettsia despite a very high frequency of tick bites in this group. PMID- 17114698 TI - Rocky mountain spotted fever in the United States, 1997-2002. AB - The increased incidence of Rocky Mountain spotted fever (RMSF) in 1997-2002 compared with previous years may be related to enhanced awareness and reporting of RMSF as well as changes in human-vector interaction. However, reports on RMSF mortality underscore the need for physician vigilance in considering a diagnosis of RMSF for febrile individuals potentially exposed to ticks and stress the importance of treating such persons regardless of the presence of a rash. PMID- 17114699 TI - Rickettsia felis in the Americas. AB - The authors describe their work in the Americas in Rickettsia felis cases in humans and the presence of Rickettsia felis in vectors. PMID- 17114700 TI - Evidence of infection in humans with Rickettsia typhi and Rickettsia felis in Catalonia in the Northeast of Spain. AB - Murine typhus is a cause of fever of intermediate duration in the south of Spain, where antibodies against Rickettsia typhi and Rickettsia felis were observed in humans. This study presents the first report from the northeast of Spain. Human serum samples were tested by serological test. R. typhi and R. felis seroprevalences were 8.8% and 3.2%, respectively. PMID- 17114701 TI - Boutonneuse fever and climate variability. AB - Researchers have long appreciated the role of climate in vector-borne diseases, including the resurgence of boutonneuse fever (BF). Portugal usually is classified as having temperate Mediterranean climate. In this new century, in analyzing the data from the Meteorology Institute, this pattern has changed and an accentuated variability in climate is being observed. BF (febre escaro nodular) is endemic and high season is from late spring and summer. The brown dog tick Rhipicephalus sanguineus. is the vector and reservoir of Rickettsia conorii complex strains: R. conorii Malish and Israeli spotted fever strain. To assess the influence of climate change in BF seasonality our aim was to compare the human sera samples received at CEVDI-INSA for laboratory diagnosis of MSF for 5 months per year from October to February, ("off-season") from 2000 to 2005. Of 1,299 sera samples in persons with suspected clinical diagnosis of MSF, 45 (3.4%) were considered positive cases and the number of positive cases has doubled in the last 2 years. BF epidemiology clearly appears to be associated with climate change, especially with low precipitation values. Physicians should be aware of increasing off-season BF cases. PMID- 17114702 TI - Brazilian spotted fever: a case series from an endemic area in southeastern Brazil: epidemiological aspects. AB - Brazilian spotted fever (BSF) is the most important tick-borne disease in Brazil and is caused by Rickettsia rickettsii and transmitted by the Ixodid tick Amblyomma cajennense, its main vector. We present epidemiologic aspects of a case series of patients admitted to the Hospital das Clinicas da UNICAMP from 1985 to 2003 with a confirmed diagnosis of BSF either by a fourfold rise in indirect immunofluorescence (IFA) titers of IgG antibodies reactive with R. rickettsii or isolation of R. rickettsii from blood or skin specimens. Seasonal variation of case occurrence seems to be associated with the life cycle of the tick. The recent reemergence of cases seems to be associated with the growing numbers of the capybara (Hydrochaeris hydrochaeris) and their expansion into urban areas. PMID- 17114703 TI - Prospective evaluation of rickettsioses in the Trakya (European) region of Turkey and atypic presentations of Rickettsia conorii. AB - In 2004 between the months of May-November, 11 patients with spotted fever group (SFG) rickettsioses were admitted to the Trakya University Hospital in Edirne, Turkey. SFG rickettsioses were diagnosed clinically. Before treatment, punch biopsy from skin lesions, especially from the eschar, was performed. Serum specimens were tested by IFA using a panel of nine rickettsial antigens, including SFG rickettsiae and R. typhi. Western blotting and standard PCR were also performed. The average age of the 11 patients (4 male and 7 female) was 51 years. All the patients had high fever; 10 (91%) had maculopapular rash; 8 (73%) had rash in the palms or on the soles. Five patients had a unique eschar; two had double eschars (64%). Two patients presented with multiple organ failure and one of them died. All the patients had significant antibody titers against SFG rickettsiae. PCR experiments of skin biopsies were positive in six (60%) of 10 skin biopsy samples and DNA sequencing of the positive PCR products gave 100% homology with Rickettsia conorii Malish 7 for opmA and gltA. Trakya Region in an endemic area for rickettsioses. In this series, three patients presented with life-threatening diseases and one of them died. This patient was the first fatal case (2.8%). Atypic and serous life-threatening presentations of rickettsioses must be kept in mind for the differential diagnosis of febrile disease in Turkey. PMID- 17114704 TI - Serologic study of rickettsioses among acute febrile patients in central Tunisia. AB - Although Mediterranean spotted or "boutonneuse" fever (MSF) has been documented in central Tunisia, other spotted fever group rickettsioses (SFGR) and typhus group rickettsioses (TGR) have received little attention in our region. We sought to determine the role of rickettsioses, Q fever, ehrlichioses, and bartonelloses among patients with acute fever. The results of this study of 47 persons with acute fever of undetermined origin are reported in this paper. We concluded that SFGR, murine typhus, and acute Q fever are common causes of acute isolate fever in summer in central Tunisia and should be investigated systematically in patients with acute fever of unknown origin. PMID- 17114705 TI - Reemergence of rickettsiosis in Oran, Algeria. AB - The presumptive cases of Mediterranean spotted fever have been identified in 1993 and since that time, its frequency has steadily increased. The prospective study, in summer 2004, was conducted in order to present the descriptive clinic and epidemiology, to identify more severe forms, the presence of the multiple eschars, and different rickettsial strains caused the disease in our region. In Oran, the cases were diagnosed clinically. In Marseille, serum specimens were tested by IFA using the panel of eight rickettsial antigen; Western blot and cross-adsorption studies were also performed in order to confirm the diagnosis. Ninety-three patients clinically diagnosed were recorded from July 3 to October 28, 2004. Eighty percent were male, the mean age was 44.3 years, 90% were exposed to dog and 32% reported tick bites. Clinical signs were as follow: presence of underlying disease (44%), sudden onset (78%), fever (100%), loss of weight (63%), conjunctivitis (43%), and a tache noire was noticed in 70%. Interestingly, two patients had two and three eschars, respectively. The rash was maculopapular (palm and sole) and purpuric in nine cases. Doxycycline was the most antibiotic (91%) with favourable outcome in 91% of the cases. Malignant form with death is reported for three patients (3.2%). Among the 93 patients, 104 serum from 65 patients were tested (serums of others patients were lost or ticket not found on tube. Sixty-three patients out of 65 had a positive serology by IFA with cross reactive antibodies especially between R. conorii, R. felis and/or R. typhi. Two others negative serology were: one precocious serum and second from the patient, which presented symptoms of MSF and tested two serums, Western blot and cross adsorption. PMID- 17114706 TI - Geoinformational mapping of foci of Siberian tick-borne rickettsiosis in Altai Krai. AB - The high rate of Siberian tick-borne rickettsiosis morbidity in Altai Krai calls for research into its causation as well as public heath concern. We constructed a thematic map to assess the medical-geographical situation, reveal disease foci, and define the risk level in the geographical regions covered. PMID- 17114707 TI - The foci of scrub typhus and strategies of prevention in the Spring in Pingtan Island, Fujian Province. AB - This study investigates the foci of tsutsugamushi disease on Pingtan Island, China, with emphasis on cases of illness that occur in the spring. The investigation of the physical and medical geography in the endemic area and detection of 141 sera samples in the population of Pingtan Island is described. The serum samples were taken from 102 soldiers and 39 residents and were tested by the indirect immunofluorescence assay. An island resident's history of illness inquiry was also conducted, and the results are discussed. We were able to determine the most prevalent types of rats and mites from the rat bodies. We isolated the DNA from the spleen of 40 rats and from five groups of mites by polymerase chain reaction. Orientia tsutsugamushi was isolated by perctoneally injecting KM mice with the patient's untreated blood along with ground rat viscera (R. losea) and ground mites (L. deliense), respectively. The isolates were typed at the molecular level. This study will then present direct prevention strategies that emphasize personal hygiene, methods of personal protection, keeping living environments clean, and it will provide strategies to eliminate rats and mites. As a result of adopting these strategies, no case of scrub typhus occurred in this region in approximately 2003-2004. PMID- 17114708 TI - Detection and identification of a novel spotted fever group rickettsia in Western Australia. AB - The extent to which rickettsiae are present in Western Australia (WA) is largely unknown. Recently there has been anecdotal evidence of a disease of unknown but possibly rickettsial origin occurring on Barrow Island, WA. Ticks were collected from people and screened using PCR. The rickettsial species was then cultured and its novelty and phylogenetic position examined. The infecting rickettsial species is divergent enough to be classified as a novel species. Sequence data suggest that the evolutionary route for Australian rickettsiae did not progress through a recent common ancestor. The pathogenic potential of the novel species is as yet unknown. PMID- 17114709 TI - Low incidence of tick-borne rickettsiosis in a Spanish Mediterranean area. AB - The aim of this study was to know the incidence of tick-borne rickettsial disease in a Mediterranean area. The incidence in 5 years for 100,000 inhabitants was 1.7 for tick-borne-lymphadenopathy (TIBOLA) and 0.4 for Mediterranean spotted fever (MSF). MSH incidence during the last few years has been lower than expected; in contrast TIBOLA seems to be an emerging disease. At the present time our data suggest a low tick-borne rickettsiosis incidence of MSF and a superior incidence of TIBOLA than MSF in the Spanish Mediterranean area. PMID- 17114710 TI - Public health problem of zoonoses with emphasis on Q fever. AB - Zoonoses are animal and human diseases. Q fever is primarily a zoonosis-an animal disease that can be transmitted to humans under certain conditions. Recent epidemiological studies suggest that Q fever should be considered as a public health problem in many countries where it is present, but unrecognizable due to inadequate disease controls. Through specific serological diagnosis of clinically suspected human Q fever cases, we are trying to determine a level of general Coxiella burnetii (C. burnetii) exposition among populations in different regions of Bosnia and Herzegovina. This would be a contribution in controlling the present and the future disease outbreaks, as well as its prevention, which is one of the prime objectives of public health. During the period from January to June 2004, in the Laboratory of the Department for Microbiology in the Medical Faculty of the University of Sarajevo, of 58 tested sera from 48 clinically suspected individuals, we confirmed the presence of specific anti-C. burnetii antibodies in 30 sera (51.7%), from 25 seropositive individuals (52.0%), by means of indirect immunofluorescent antibody (IFA) testing. Urgent steps must be taken in public education to help decrease the risk of C. burnetii infection among at-risk populations in regions of Bosnia and Herzegovina. PMID- 17114711 TI - Rickettsia slovaca infection: DEBONEL/TIBOLA. AB - This study describes the epidemiological, clinical, and microbiological characteristics of a new tick-borne disease in Spain-Dermacentor-borne necrosis erythema lymphadenopathy (DEBONEL). The clinical presentations include an eschar at the site of the tick bite, surrounded by an erythema and painful regional lymphadenopathy. The disease appears during the colder months and its vector is Dermacentor marginatus (D. marginatus). From January 1990 to December 2004, 54 patients presented at Hospital of La Rioja with these clinical and epidemiological data. The ratio of females to males was 32/22. The average age was 37 years. In all cases tick bites were located on the upper body (90% on the scalp). The median incubation period was 4.7 days. Signs and symptoms were mild in all cases. Only a small number of patients presented mild and nonspecific abnormalities in a complete blood cell count and mild elevation of erythrocyte sedimentation rates and C-protein reactive and liver enzyme levels. Serological evidence of acute rickettsiosis was observed in 19 patients (61%). In 29% sera tested by polymerase chain reactions (PCRs) were positive. The sequence obtained from a PCR product revealed 98% identity with Rickettsia sp. strains RpA4, DnS14, and DnS28. All ticks removed from patients were PCR-positive. Sequencing showed 8 of them identified as R. slovaca and 2 as Rickettsia sp. strains RpA4, DnS14, and DnS28. PMID- 17114712 TI - Infective endocarditis due to Bartonella spp. and Coxiella burnetii: experience at a cardiology hospital in Sao Paulo, Brazil. AB - Bartonella spp. and Coxiella burnetii are recognized as causative agents of blood culture-negative endocarditis (BCNE) in humans and there are no studies of their occurrences in Brazil. The purpose of this study is to investigate Bartonella spp. and C. burnetii as a causative agent of culture-negative endocarditis patients at a cardiology hospital in Sao Paulo, Brazil. From January 2004 to December 2004 patients with a diagnosis of endocarditis at our Institute were identified and recorded prospectively. They were considered to have possible or definite endocarditis according to the modified Duke criteria. Those with blood culture-negative were tested serologically using the indirect immunofluorescent assay (IFA) for Bartonella henselae, B. quintana, and C. burnetii. IFA-IgG titers >800 for Bartonella spp. and C. burnetii were considered positive. A total of 61 patients with endocarditis diagnosis were evaluated, 17 (27%) were culture negative. Two have had IgG titer greater than 800 (>/=3,200) against Bartonella spp. and one against C. burnetii (phase I and II>/=6,400). Those with Bartonella induced endocarditis had a fatal disease. Necropsy showed calcifications and extensive destruction of the valve tissue, which is diffusely infiltrated with mononuclear inflammatory cells predominantly by foamy macrophages. The patient with C. burnetii endocarditis received specific antibiotic therapy. Reports of infective endocartitis due to Bartonella spp. and C. burnetii in Brazil reveal the importance of investigating the infectious agents in culture-negative endocarditis. PMID- 17114713 TI - Arthropod-borne diseases in homeless. AB - Homeless people are particularly exposed to ectoparasite. The living conditions and the crowded shelters provide ideal conditions for the spread of lice, fleas, ticks, and mites. Body lice have long been recognized as human parasites and although typically prevalent in rural communities in upland areas of countries close to the equator, it is now increasingly encountered in developed countries especially in homeless people or inner city economically deprived population. Fleas are widespread but are not adapted to a specific host and may occasionally bite humans. Most common fleas that parasite humans are the cat, the rat, and the human fleas, Ctenocephalides felis, Xenopsylla cheopis, and Pulex irritans, respectively. Ticks belonging to the family Ixodidae, in particular, the genera Dermacentor, Rhipicephalus, and Ixodes, are frequent parasites in humans. Sarcoptes scabiei var. hominis is a mite (Arachnida class) responsible for scabies. It is an obligate parasite of human skin. The hematophagic-biting mite, Liponyssoides sanguineus, is a mite of the rat, mouse, and other domestic rodents but can also bite humans. Finally, the incidence of skin disease secondary to infestation with the human bedbug, Cimex lectularius, has increased recently. Bacteria, such as Wolbacchia spp. have been detected in bedbug. The threat posed by the ectoparasite in homeless is not the ectoparasite themselves but the associated infectious diseases that they may transmit to humans. Except for scabies all these ectoparasites are potential vectors for infectious agents. Three louse-borne diseases are known at this time. Trench fever caused by Bartonella quintana (B. quintana), epidemic typhus caused by Rickettsia prowazekii, and relapsing fever caused by the spirochete Borrelia recurrentis. Fleas transmit plague (Xenopsylla cheopis and Pulex irritans), murine typhus (Xenopsylla cheopis), flea-borne spotted rickettsiosis on account of the recently described species Rickettsia felis (C. felis), and occasionally cat scratch disease on account of Bartonella henselae (C. felis). The role of fleas as potential vector of B. quintana has recently been suggested. Among the hematophagic-biting mites, L. sanguineus, is responsible for the transmission of Rickettsia akari, the etiologic agent of rickettsialpox. Virtually, no data are available on tick-borne disease in this population. This article will deal with epidemiology, diagnosis, prevention, and treatment of these ectoparasite and the infectious diseases they transmit to the homeless people. PMID- 17114714 TI - Clinical diagnosis and treatment of human granulocytotropic anaplasmosis. AB - Tick-borne rickettsiae in the genera Ehrlichia and Anaplasma are intracellular bacteria that infect wild and domestic mammals and, more recently, man. The increased desire of humans for recreational activities outdoors has increased the exposure to potential human pathogens that previously cycled almost exclusively within natural, nonhuman enzootic hosts. Anaplasma phagocytophilum causes an acute, nonspecific febrile illness of humans previously known as human granulocytotropic ehrlichiosis (HGE) and now called human granulocytotropic anaplasmosis (HGA). The first patient to have recognized HGA was hospitalized at St Mary's Hospital in Duluth, Minnesota, USA in 1990. However, the clinical and laboratory presentation of this infection remained undefined until 1994, when Bakken and collaborators published their experience with 12 patients who had HGA. By the end of December 2004, at least 2,871 cases of HGA had been reported from 13 U.S. states to the Centers for Disease Control and Prevention (CDC). A limited number of laboratory-confirmed cases have been reported from countries in Europe, including Austria, Italy, Latvia, the Netherlands, Norway, Poland, Slovenia, Spain, and Sweden. Ixodes persulcatus-complex ticks are the arthropod hosts for Borrelia burgdorferi, the agent of Lyme borreliosis, and are also the arthropod hosts for A. phagocytophilum. Most cases of HGA have been contracted in geographic regions that are endemic for Lyme borreliosis. Male patients outnumber female patients by a factor of 3 to 1 and as many as 75% of patients with HGA have had a tick bite prior to their illness. Seroepidemiologic studies have demonstrated that HGA for the most part is a mild or even asymptomatic illness. However, older individuals and patients who are immunocompromised by natural disease processes or medications may develop an acute, influenza-like illness characterized by high fever, rigors, generalized myalgias, and severe headache. Local skin reactions at the site of the tick bite have not been described, and nonspecific skin rashes have been reported only occasionally. Anaplasmosis is associated with variable but suggestive changes in routine laboratory test parameters. Most patients develop transient reductions in total leukocyte and platelet concentrations. Relative granulocytosis accompanied by a left shift and lymphopenia during the first week of illness has been reported frequently. Serum hepatic transaminase concentrations usually increase two- to fourfold, and inflammatory markers, such as C-reactive protein and the erythrocyte sedimentation rate, rise during the acute phase. Abnormal laboratory findings may return toward normal range for patients who have been ill for more than 7 days, which may obfuscate the clinical decision making. Characteristic clusters of bacteria (morulae) are observed in the cytoplasm of peripheral blood granulocytes in 20% to 80% of infected patients during the acute phase of illness. The clinical diagnosis may be confirmed retrospectively by specific laboratory tests, which include positive polymerase chain reaction (PCR), identification of A. phagocytophilum in culture of acute-phase blood, or the detection of specific antibodies to A. phagocytophilum in convalescent serum. Virtually all patients have developed serum antibodies to A. phagocytophilum after completion of antibiotic therapy, and demonstration of seroconversion by indirect immunofluorescent antibody testing of acute-phase and convalescent-phase serum samples is currently the most sensitive and specific tool for laboratory confirmation of HGA. Treatment with doxycycline usually results in rapid improvement and cure. Most patients with HGA have made an uneventful recovery even without specific antibiotic therapy. However, delayed diagnosis in older and immunocompromised patients may place those individuals at risk for an adverse outcome, including death. Thus, prompt institution of antibiotic therapy is advocated for any patient who is suspected to have HGA and for all patients who have confirmed HGA. PMID- 17114715 TI - Diagnosis of Coxiella burnetii pericarditis using a systematic prescription kit in case of pericardial effusion. AB - Coxiella burnetii, regarded as a potential agent of pericarditis, wa found to be responsible for almost 5% of the cases of idiopathic pericardial effusion reported in this series. Diagnosis was aided by use of a systematic kit described in this paper. PMID- 17114716 TI - Brazilian spotted fever: a case series from an endemic area in southeastern Brazil: clinical aspects. AB - This case series study is based on a retrospective review of medical records and case notification files of patients admitted to The Hospital das Clinicas da UNICAMP from 1985 to 2003 with a confirmed diagnosis of BSF either by fourfold rise in indirect immunofluorescence assay (IFA) titers of IgG antibodies reactive with R. rickettsii or isolation of R. rickettsii from blood or skin specimens. A median lethality of 41.9 % was observed between 1985 and 2004. The case-fatality ratio of 30 % in our study, lower than the overall Sao Paulo state ratio, could be explained by a higher index of suspicion and a larger experience in our hospital, a regional referral center for BSF. The presence of the classical triad of fever, rash, and headache as described in RMSF was observed in fever than half (35.2%) of our patients. PMID- 17114717 TI - Revisiting brazilian spotted fever focus of Caratinga, Minas Gerais State, Brazil. AB - We revisited a Brazilian spotted fever focal area in Minas Gerais state, Brazil, in 2002, and performed a serologic survey in dogs and cats. The results of this survey are compared with the survey made 10 years before. The possible efficacy of vector control measures adopted in this area and the role of dogs and horses as sentinels of infection by Rickettsia are discussed. PMID- 17114718 TI - Fatal case of Brazilian spotted fever confirmed by immunohistochemical staining and sequencing methods on fixed tissues. AB - The authors describe the first characterization of Rickettsia rickettsii in a fatal case occurring in Rio de Janeiro State, Brazil. PMID- 17114719 TI - Detection of Rickettsia rickettsii and Rickettsia sp. in blood clots in 24 patients from different municipalities of the State of Sao Paulo, Brazil. AB - The authors detected Rickettsia genus organisms using shell vial and polymerase chain reaction (PCR)/sequencing analysis in blood clots in patients suspected of having Brazilian spotted fever (BSF). DNA was detected using PCR with three sets of primers to access the gltA, ompA, and ompB genes. Sequence analysis was carried out using an automatic sequencer with Bioedit software. Seventy-five percent of the culture samples were positive and all samples amplified rickettsial gene fragments. To date, 46% of the samples have been sequenced. PMID- 17114720 TI - Mediterranean spotted fever in crete, Greece: clinical and therapeutic data of 15 consecutive patients. AB - The clinical, epidemiological, and therapeutic aspects of 15 patients with Mediterranean spotted fever (MSF), admitted to the Internal Medicine Department of the General Hospital of Sitia (southeastern Crete, Greece) between December 2000 and July 2003, were studied. Diagnosis was made on the basis of clinical signs and symptoms and was confirmed by serology. Of the patients studied, 67% were men and 33% women, with a median age of 52 years (range of 23-76 years). Ten cases (67%) were diagnosed between May and July. Of all the patients, 93% had a history of contact with animals, mainly with sheep (11 patients, 73%), while 53% of them had a history of tick-bite (33%), or reported the presence of ticks in their environment (20%). The typical eschar lesion (tache noir) at the tick-bite site was present in 53% of the patients, while the rash was present in 87% of them. Laboratory findings included leukopenia (47%), thrombocytopenia (54%), elevation of transaminases (80%), hyponatremia (33%), and microscopic hematuria (80%). Four patients (27%) displayed pulmonary infiltrates on chest radiography. All patients were treated with doxycycline (200 mg daily) and recovered rapidly. Renal function deteriorated in one patient with chronic renal failure, but he recovered thereafter. PMID- 17114721 TI - Prevalence of rickettsia felis-like and Bartonella Spp. in Ctenocephalides felis and Ctenocephalides canis from La Rioja (Northern Spain). AB - Our aim was to determine the presence of Rickettsia spp. and Bartonella spp. in Ctenocephalides felis and Ctenocephalides canis from La Rioja (Spain). A total of 88 specimens were tested by polymerase chain reaction (PCR) using gltA and ompB genes as targets for Rickettsia spp., and 16S rRNA and ribC genes for Bartonella spp. Rickettsia felis-like (28.4%), Bartonella clarridgeiae (6.8%), and Bartonella henselae (3.4%) were detected in Ctenocephalides spp. Other Bartonella sp. different from B. clarridgeiae and B. henselae could also be present in fleas from La Rioja. PMID- 17114722 TI - Prediction of habitat suitability for ticks. AB - This article offers a personal perspective on the efforts to model the habitat suitability for ticks. A serious effort is being made to provide maps at an adequate resolution that can plot the probability of the presence and even the abundance of tick parasites of humans and that can be used to develop control programs for species with economic interest. Some of the methods being currently evaluated are described together with a summary of the techniques using remotely sensed information to capture the abiotic features of the habitat. Some examples, developed using different methods, are provided for the Mediterranean region and part of the U.S., together with an overview of the performance of these models. PMID- 17114723 TI - Natural infection, transovarial transmission, and transstadial survival of Rickettsia bellii in the Tick Ixodes loricatus (Acari: Ixodidae) from Brazil. AB - An Ixodes loricatus engorged female, infected with Rickettsia bellii, was collected from an opossum (Didelphis aurita) in Mogi das Cruzes, Sao Paulo State, Brazil. Two consecutive laboratory tick generations (F(1) and F(2)) reared from this single engorged female were evaluated for Rickettsia infection by polymerase chain reaction (PCR) targeting specific Rickettsia genes. Immature ticks fed on naive Wistar rats (Rattus norvegicus) and adult ticks fed on opossum (D. aurita), both free of ticks and rickettsial infection. PCR performed on individual ticks from the F(1) (20 larvae, 10 nymphs, and 10 adults) and the F(2) (30 larvae, 30 nymphs, and 15 adults) yielded expected bands compatible with Rickettsia. All the PCR products that were sequenced, targeting gltA gene, resulted in sequences identical to each other and 99.7% (349/350) similar to the corresponding sequence of R. bellii in GenBank. The R. bellii infection on ticks from the second laboratory generation (F(2)) was confirmed by other PCR protocols and successful isolation of R. bellii in cell culture. We report for the first time a Rickettsia species infecting I. loricatus, and the first report of R. bellii in the tick genus Ixodes. We conclude that there was an efficient transovarial transmission and transstadial survival of this Rickettsia species in the tick I. loricatus. Our results suggest that R. bellii might be maintained in nature solely by transovarial transmission and transstadial survival in ticks (no amplifier vertebrate host is needed), since there has been no direct or indirect evidence of infection of vertebrate hosts by R. bellii. PMID- 17114724 TI - Prevalence of bacterial agents in Ixodes persulcatus ticks from the Vologda Province of Russia. AB - The prevalence of rickettsiae, ehrlichiae, and the rickettsia-like endosymbiont called Montezuma relative to that of Borrelia was determined in questing Ixodes persulcatus (I. persulcatus) ticks collected in 2002-2003 from Vologda Province, Russia. Ehrlichia muris, Anaplasma phagocytophilum, Montezuma, and new spotted fever group rickettsiae were detected by polymerase chain reaction (PCR) for the first time in this area. The rickettsiae were all Candidatus Rickettsia tarasevichiae, the furthest west this organism has been detected. After Borrelia, Montezuma was the agent most frequently detected; it may be present throughout the distribution of I. persulcatus in Russia. Ehrlichiae and rickettsiae frequently share the same tick host with Borrelia burgdorferi sensu lato so cotransmission and mixed infections in vertebrate hosts, including humans, may occur. PMID- 17114725 TI - Ecology and molecular epidemiology of tick-borne rickettsioses and anaplasmoses with natural foci in Russia and Kazakhstan. AB - During our more than 20 years of monitoring, we have used epidemiological, field, and experimental methods for characterization of natural foci of tick-borne rickettsioses in Russia. The main results were obtained through genetic methods (PCR sequence) at the Universite de la Mediterranee (Marseille, France). We describe considerable heterogeneity of tick-borne alpha(1)-proteobacteria: 16 microorganisms the of the order Rickettsiales were detected in Russia and Kazakhstan. R. sibirica-caused North Asiatic tick-borne rickettsiosis is the main tick-borne rickettsiosis in Russia, with wide distribution in Siberia and the Russian Far East and high epidemic activity of natural foci of different landscape types. Our results show circulation of different pathogenic rickettsiae in the same endemic territories. In the Far East region, R. sibirica subsp. R. sibirica, R. sibirica subsp. BJ-90, and R. heilongjiangensis were detected; in the Altay and Krasnojarsk regions, R. sibirica subsp. R. sibirica and R. heilongjiangensis; and in the Kurgan district of West Siberia, R. sibirica subsp. R. sibirica and R. slovaca. The roles of more than 15 new genotypes of alpha(1) proteobacteria in infectious disease in Russia and Kazakhstan are in need of further study. PMID- 17114726 TI - Prevalence of Rickettsia felis in Ctenocephalides felis and Ctenocephalides canis from Uruguay. AB - Our aim was to determine the presence of Rickettsia spp. in 66 fleas from Uruguay. Rickettsial DNA was amplified using gltA and ompB PCR primers. Rickettsia spp. were found in 41% of the fleas (25 Ctenocephalides felis and 2 Ctenocephlides canis). Sequences resulted in the identification of Rickettsia felis and four genotypes closely related to this species (Rickettsia sp. TwKM03, California 2, Hf187, and RF2125). The presence of R. felis in fleas from Uruguay in was demonstrated. This is the second species of Rickettsia identified in Uruguay in the past 2 years using molecular approaches, and it is helping to clarify the etiology of rickettsial diseases in the region. PMID- 17114727 TI - Highly variable year-to-year prevalence of Anaplasma phagocytophilum in Ixodes ricinus ticks in northeastern Poland: a 4-year follow-up. AB - Anaplasma phagocytophilum is transmitted mainly by Ixodes ricinus ticks in Europe. We followed-up A. phagocytophilum infection rate in I. ricinus in three selected collection sites in northeastern Poland during a four-year period. Overall infection rate was 14.1% (208/1474) with highest infection rate among females (36.8% verses males 8.2% and nymphs 0.9%). We noted a very big year-to year variation of infection prevalence in each collection cite every year reflecting changeable granulocytic anaplasmosis risk for humans and animals. PMID- 17114728 TI - Detection of Anaplasma phagocytophilum, Coxiella burnetii, Rickettsia spp., and Borrelia burgdorferi s. l. in Ticks, and wild-living animals in western and middle Slovakia. AB - In this study, three tick species (Ixodes ricinus, Dermacentor marginatus, and D. reticulatus), small terrestrial mammals, and game were examined by PCR for the presence of tick-borne pathogens Anaplasma phagocytophilum, Coxiella burnetii, Rickettsia spp., and Borrelia burgdorferi Sensu lato. PMID- 17114729 TI - Prevalence of Anaplasma phagocytophilum, Rickettsia sp. and Borrelia burgdorferi sensu lato DNA in questing Ixodes ricinus ticks from France. AB - A total of 4701 Ixodes ricinus, collected during the summer of 2003, were analyzed for three pathogens. DNA was detected from the three pathogens. Co detection of more than one pathogen was observed. PMID- 17114730 TI - Prevalence of spotted fever group Rickettsia species detected in ticks in La Rioja, Spain. AB - Our objective was to learn the prevalence of spotted fever group (SFG) Rickettsia detected in ticks in La Rioja, in the north of Spain. From 2001 to 2005, 496 ticks representing 7 tick species were analysed at the Hospital de La Rioja. Ticks were removed from humans with or without rickettsial syndrome (n = 59) or collected from mammals (n = 371) or from vegetation by dragging (n = 66). The presence of SFG Rickettsia in these ticks was investigated by semi-nested PCR (ompA gene) and sequencing. A phylogenetic tree using Clustal method (neighbor joining) was constructed with these data. Only 3 of 170 Hyalomma marginatum ticks carried SFG Rickettsia. Sequencing analysis demonstrated the presence of Rickettsia aeschlimannii (1.8%). Furthermore, Rickettsia massiliae and BAR29 were found in 3 of 120 Rhipicephalus sanguineus specimens (2.5%). In contrast, 81 of 83 tested Dermacentor marginatus ticks were PCR-positive (97%). Rickettsia slovaca (40.6%) and Rickettsia sp. strains RpA4, DnS14, DnS28 and JL-02 (59.3%) were found within this tick species. No SFG Rickettsia was detected using ompA primers when Ixodes ricinus, Rhipicephalus bursa, Rhipicephalus turanicus, Rhipicephalus eversti eversti, Hyalomma detritum scupense and Rhipicephalus sp. were analyzed. We detected 17.5% of ticks associated with different SFG Rickettsia: R. aeschlimannii, R. massiliae, BAR29, R. slovaca and Rickettsia sp. strains RpA4, DnS14, DnS28 and JL-02. Their presence has to be taken into account since most of them have been recognized as human pathogens. PMID- 17114731 TI - Prevalence of Rickettsia slovaca in Dermacentor marginatus ticks removed from wild boar (Sus scrofa) in northeastern Spain. AB - Rickettsia slovaca, the causative agent of TIBOLA, is transmitted by Dermacentor ticks. Dermacentor marginatus is the most widely species distributed in northeastern Spain, and the wild boar constitutes the main host. D. marginatus ticks were collected from hunter-killed wild boar and were tested by PCR/RFLP. Rickettsial DNA-positive ticks were sequenced using the ompA PCR primers. The prevalence of R. slovaca in D. marginatus ticks was 17.7%. Other spotted fever group rickettsiae were detected in ticks, but these were not definitely identified. PMID- 17114732 TI - Prevalence data of Rickettsia slovaca and other SFG Rickettsiae species in Dermacentor marginatus in the southeastern Iberian peninsula. AB - In southern Spain, Dermacentor marginatus ticks can be infected with several genospecies of spotted fever Group (SFG) Rickettsia. We developed a nested polymerase chain reaction assay by using a species-specific probe targeting the ompA gene to detect and differentiate between the two groups of rickettsiae previously described in D. marginatus. SFG rickettsia has been detected in 85.15% of ticks studied (26.7% of positives have been to R. slovaca, the causative agent of TIBOLA-DEBONEL, and 73.3% to SFG rickettsia closely related to strains RpA4-JL 02-DnS14-DnS28). PMID- 17114733 TI - Spotted fever group rickettsiae in ticks feeding on humans in northwestern Spain: is Rickettsia conorii vanishing? AB - During a 7-year study, we identified and analyzed by PCR 4,049 ticks removed from 3,685 asymptomatic patients in Castilla y Leon (northwestern Spain). A total of 320 ticks (belonging to 10 species) were PCR-positive for rickettsiae. Comparison of amplicon sequences in databases enabled us to identify eight different spotted fever group (SFG) rickettsiae: Rickettsia slovaca, Rickettsia sp. IRS3/IRS4, R. massiliae/Bar29, R. aeschlimannii, Rickettsia sp. RpA4/DnS14, R. helvetica, Rickettsia sp. DmS1, and R. conorii. Although Mediterranean spotted fever (MSF) is an endemic disease in Castilla y Leon, R. conorii was found in only one Rhipicephalus sanguineus tick, whereas other pathogenic SFG rickettsiae were much more prevalent in the same area. Our data suggest that in Castilla y Leon, many MSF or MSF-like cases attributed to R. conorii could have been actually caused by other SFG rickettsiae present in ticks biting people in this region of Spain. PMID- 17114734 TI - A rickettsial mixed infection in a Dermacentor variabilis tick from Ohio. AB - We present the first report of superinfection in a Dermacentor variabilis tick from nature. The single tick, collected in Ohio, was found infected with Rickettsia belli, R. nontanensis, and R. rickettsii. PMID- 17114735 TI - Rocky Mountain spotted fever in Arizona: documentation of heavy environmental infestations of Rhipicephalus sanguineus at an endemic site. AB - A recent epidemiologic investigation identified 16 cases and 2 deaths from Rocky Mountain spotted fever (RMSF) in two eastern Arizona communities. Prevalence studies were conducted by collecting free-living ticks (Acari: Ixodidae) from the home sites of RMSF patients and from other home sites within the community. Dry ice traps and flagging confirmed heavy infestations at many of the home sites. Only Rhipicephalus sanguineus ticks were identified and all developmental stages were detected. It is evident that under certain circumstances, this species does transmit Rickettsia rickettsii to humans and deserves reconsideration as a vector in other geographic areas. PMID- 17114736 TI - An outbreak of Rocky Mountain Spotted Fever associated with a novel tick vector, Rhipicephalus sanguineus, in Arizona, 2004: preliminary report. AB - This study describes preliminary results of an investigation of RMSF in Arizona associated with the brown dog tick, Rhipicephalus sanguineus. High numbers of dogs and heavy infestations of ticks created a situation leading to human disease. PMID- 17114737 TI - Incidence and distribution pattern of Rickettsia felis in peridomestic fleas from Andalusia, Southeast Spain. AB - The presence of Rickettsia felis was investigated in three species of pulicid fleas (Ctenocephalides felis, Ctenocephalides canis and Pulex irritans) collected in 38 locales in Andalusia (Spain) over the period 1999-2004. Amplification of a fragment of OmpB gene was positive in 54.17 % of lots of Ct. felis. The identity of the PCR bands was confirmed as R. felis by sequence data obtained directly from the PCR amplicon. No rickettsia was found in Ct. canis nor P. irritans. PMID- 17114738 TI - Molecular identification of Rickettsia felis-like bacteria in Haemaphysalis sulcata ticks collected from domestic animals in southern Croatia. AB - Haemaphysalis sulcata ticks collected from sheep and goats in southern Croatia were found infected with rickettsiae. Molecular analysis of the complete gltA gene and portion of 17 kDa and ompB genes revealed the presence of Rickettsia felis-like bacteria in up to 26% of tested ticks. PMID- 17114739 TI - Expression of rOmpA and rOmpB protein in Rickettsia massiliae during the Rhipicephalus turanicus life cycle. AB - Rhipicephalus turanicus tick colony infected in the laboratory with Rickettsia massiliae showed that the rickettsia is transovarially and transdatially tramsmitted. The expression of rOmpB did not change with temperature or the stages of the tick life cycle. In contrast, rOmpA was less expressed during the larval stage. PMID- 17114740 TI - Lice infestation and lice control remedies in the Ukraine. AB - A permanent decrease was seen in the prevalence of lice infestation among population of the Ukraine from 1990-2004. The prevalence of lice infestation among children under 14 years of age was 6-27 times more than that in adults. The highest figures were among children 7-14 years old. During all of the observation period there were changing tendencies relative to the groups. The greatest number of cases of infection with lice were noted in the months when the control inspections of children in education institutions were performed. PMID- 17114741 TI - Prevalence of Rickettsia felis in the fleas Ctenocephalides felis felis and Ctenocephalides canis from two Indian villages in Sao Paulo Municipality, Brazil. AB - We evaluated the presence of Rickettsia infection among fleas collected on domestic dogs in two Guarani Indian communities in the suburban area of Sao Paulo Municipality, Brazil. A total of 114 Ctenocephalides felis felis and 47 Ctenocephalides canis were collected from 40 dogs. A total of 41 C. felis felis (36.0%) and 9 C. canis (19.1%) fleas yielded expected bands by PCR, which were all shown by DNA sequencing to be indentical to the corresponding sequence of a fragment of the Rickettsia felis gltA gene deposited in GenBank. The overall prevalence of R. felis was 31.0% (49/161). PMID- 17114742 TI - Population survey of Egyptian arthropods for rickettsial agents. AB - Between June 2002 and July 2003, 987 fleas, representing four species, and 1019 ticks, representing one argasid and eight ixodid species, were collected from Egyptian animals. These arthropods were tested for rickettsial agents using polymerase chain reaction. DNAs from Anaplasma and Ehrlichia spp. were detected in 13 ticks. Previously undescribed Bartonella spp. were detected in 21 fleas. Coxiella burnetii was detected in two fleas and 20 ticks. Rickettsia typhi was detected in 27 fleas from 10 cities. Spotted fever group rickettsiae were detected in both fleas and ticks and included Rickettsia aeschlimanii and an unnamed Rickettsia sp. PMID- 17114743 TI - First molecular detection of R. conorii, R. aeschlimannii, and R. massiliae in ticks from Algeria. AB - Ticks collected in Northern Algeria between May 2001 and November 2003 were tested by PCR for the presence of Rickettsia spp. DNA using primer amplifying gltA and OmpA genes. Three different spotted fever group rickettsias were amplified from these ticks: R. Conorii subsp. P. conorii strain Malish in Rhipicephalus sanguineus, R. aeschlimannii in Hyalomma marginatum, and R. massiliae in Rhipicephalus turanicus. Our results confirm the presence of R. conorii in ticks in Algeria and provide the first detection of R. aeschlimannii and R. massiliae in Algeria. PMID- 17114744 TI - Ornithodoros moubata, a soft tick vector for Rickettsia in east Africa? AB - Omithodoros moubata complex (Argasidae) ticks collected from human dwellings in central Tanzania were found to carry a novel rickettsial species that clustered among the spotted fever group. Although no evidence of human infection was evident, these ticks feed primarily on man, thus providing opportunity for zoonotic infection. PMID- 17114745 TI - Detection of members of the genera Rickettsia, Anaplasma, and Ehrlichia in ticks collected in the Asiatic part of Russia. AB - A total of 395 adult ixodid ticks from three genera (Dermacentor, Haemaphysalis, and Ixodes) collected from the Urals to the Far East of Russia were tested by PCR and sequencing for the presence of spotted fever rickettsiae, anaplasmae, and ehrlichiae. Four, pathogens recognized in humans were detected in ticks: Rickettsia sibirica, R. heilongjiangensis, R. helvetica, and Anaplasma phagocytophilum. In addition, rickettsiae and ehrlichiae of unknown pathogenicity were detected, including Rickettsia sp. RpA4, Rickettsia sp. DnS14, Rickettsia sp. DnS28, "Candidatus R. tarasevichiae," a rickettsia closely related to R. helvetica, A. bovis, Ehrlichia muris, "Ehrlichia-like" "Schotti variant," and bacterium "Montezuma." Our findings indicated the distribution of rickettsiae and ehrlichiae in hard ticks in Russia. PMID- 17114746 TI - Characterization of Dermacentor variabilis molecules associated with Rickettsial infection. AB - To ultimately define the virulence factors of rickettsiae, an understanding of the biology of the organism is essential. Comprehension of the pathogen-human interaction is critical to the development of control measures; and, in the case of vector-borne diseases, the role of the vector in maintaining and transmitting pathogens to vertebrate hosts is crucial to ultimate control. Recent studies have identified tick molecules that are likely involved in the tick-rickettsiae interchange, including tick response to infection and possible molecules exploited by rickettsiae during transmission events. We have further characterized several tick-derived molecules, including a histamine release factor, serine proteases, and lysozymes. PMID- 17114747 TI - Ticks, tick-borne rickettsiae, and Coxiella burnetii in the Greek Island of Cephalonia. AB - Domestic animals are the hosts of several tick species and the reservoirs of some tick-borne pathogens; hence, they play an important role in the circulation of these arthropods and their pathogens in nature. They may act as vectors, but, also, as reservoirs of spotted fever group (SFG) rickettsiae, which are the causative agents of SFG rickettsioses. Q fever is a worldwide zoonosis caused by Coxiella burnetii (C. burnetii), which can be isolated from ticks. A total of 1,848 ticks (954 female, 853 male, and 41 nymph) were collected from dogs, goats, sheep, cattle, and horses in 32 different localities of the Greek island of Cephalonia. Rhipicephalus (Rh.) bursa, Rh. turanicus, Rh. sanguineus, Dermacentor marginatus (D. marginatus), Ixodes gibbosus (I. gibbosus), Haemaphysalis (Ha.) punctata, Ha. sulcata, Hyalomma (Hy.) anatolicum excavatum and Hy. marginatum marginatum were the species identified. C. burnetii and four different SFG rickettsiae, including Rickettsia (R.) conorii, R. massiliae, R. rhipicephali, and R. aeschlimannii were detected using molecular methods. Double infection with R. massiliae and C. burnetii was found in one of the positive ticks. PMID- 17114748 TI - Molecular characterization of Rickettsia rickettsii infecting dogs and people in North Carolina. AB - Rocky Mountain spotted fever (RMST) is an important cause of morbidity and mortality in people and dogs in the United States. Disease manifestations are strikingly similar in both species, and illness in dogs can precede illness in people. R. rickettsii has been identified as a Select Agent by the CDC as a Category C priority pathogen by the National Institute of Allergic and Infectious Diseases because it is amenable to use as a bioterror agent. The clinical and temporal relationship of naturally occurring diseases in dogs and people suggests that dogs could serve as sentinels for natural infection and bioterrorist attacks using this organism. Recognizing genetic modifications in naturally occurring disease agents in order to distinguish them from intentionally released agents are priorities put forth by the NIAID. To determine whether the rickettsiae naturally infecting dogs is the same as those that infect persons in a given geographical region, we characterized rickettsial isolates obtained from three dogs and two persons diagnosed with RMSF in North Carolina. Portions of three genes (ompA, rrs, and gltA) amplified by PCR were cloned and sequenced or directly sequenced. Reactions were run in duplicate in forward and reverse directions. Gene sequences were aligned with known sequences deposited in GenBank and with each other. Sequences of the 5' region of the ompA gene were 100% homologous with a tick strain (Bitterroot) of R. rickettsii for all five isolates. Sequences of the rrs gene were 99.8 99.9% homologous with a tick strain (Sawtooth) of R. rickettsii. rrs gene sequences from one dog and the two persons was identical. Sequences of one dog isolate differed from these by one base pair. Sequences from another dog isolate differed by two base pairs. Sequences of the gltA gene are pending. This confirms on a molecular level that R. rickettsii causing naturally occurring RMSF in dogs in North Carolina is highly homologous to R. rickettsii that causes the disease in people in the same region. Sequence data will be deposited in GenBank, thereby providing genetic information regarding naturally occurring R. rickettsii. PMID- 17114749 TI - Bartonella infection in domestic cats and wild felids. AB - Bartonella are vector-borne, fastidious Gram-negative bacteria causing persistent bacteremia in their reservoir hosts. Felids represent a major reservoir for several Bartonella species. Domestic cats are the main reservoir of B. henselae, the agent of cat-scratch disease. Prevalence of infection is highest in warm and humid climates that are optimal for the survival of cat fleas, as fleas are essential for the transmission of the infection. Flea feces are the likely infectious substrate. Prevalence of B. henselae genotypes among cat populations varies worldwide. Genotype Houston I is more prevalent in the Far East and genotype Marseille is dominant in western Europe, Australia, and the western United States. Cats are usually asymptomatic, but uveitis, endocarditis, neurological signs, fever, necrotic lesions at the inoculation site, lymphadenopathy, and reproductive disorders have been reported in naturally or experimentally infected cats. Domestic cats are also the reservoir of B. clarridgeiae and co-infection has been demonstrated. B. koehlerae has been isolated from domestic cats, and was identified in cat fleas and associated with a human endocarditis case. B. bovis was isolated from a few cats in the United States and B. quintana DNA was recently identified in a cat tooth. Bartonella spp. have also been isolated from free-ranging and captive wild felids from North America and Africa. Whereas, B. henselae was identified in African lions and a cheetah, some strains specific to these wild cats have also been identified, leading to the concept of a B. henselae group including various subspecies, as previously described for B. vinsonii. PMID- 17114750 TI - Anaplasmosis: focusing on host-vector-pathogen interactions for vaccine development. AB - Anaplasma marginale and A. phagocytophylum are intracellular rickettsiae that cause bovine anaplasmosis and human granulocytic anaplasmosis, respectively. The ultimate vaccine for the control of anaplasmosis would be one that reduces infection and transmission of the pathogen by ticks. Effective vaccines for control of anaplasmosis are not available despite attempts using different approaches, such as attenuated strains, infected erythrocyte and tick cell derived purified antigens, and recombinant pathogen and tick-derived proteins. Three lines of functional analyses were conducted by our laboratory to characterize host-tick-Anaplasma interactions to discover potential vaccine candidate antigens to control tick infestations and the infection and transmission of Anaplasma spp.: (1) characterization of A. marginale adhesins involved in infection and transmission of the pathogen, (2) global expression analysis of genes differentially expressed in HL-60 human promyelocytic cells in response to infection with A. phagocytophilum, and (3) identification and characterization of tick-protective antigens by expression library immunization (ELI) and analysis of expressed sequence tags (EST) in a mouse model of tick infestations and by RNA interference in ticks. These experiments have resulted in the characterization of the A. marginale MSP1a as an adhesin for bovine erythrocytes and tick cells, providing support for its use as candidate vaccine antigen for the control of bovine . Microarray analysis of genes differentially expressed in human cells infected with A. phagocytophilum identified key molecules involved in pathogen infection and multiplication. The screening for tick-protective antigens resulted in vaccine candidates reducing tick infestation, molting, and oviposition and affecting Anaplasma infection levels in ticks. PMID- 17114751 TI - Evaluation of E. ruminantium genes in DBA/2 mice as potential DNA vaccine candidates for control of heartwater. AB - Heartwater caused by the rickettsia Ehrlichia ruminantium (E. ruminantium) is an acute and fatal tick-borne disease of domestic and some wild ruminants. A user friendly vaccine does not exist. We selected and tested nine genes of E. ruminantium for protection against challenge in a DBA/2 mouse model, in order to identify candidate genes for incorporation into a recombinant vaccine. Of the nine DNA vaccine constructs tested, four DNA constructs 14HWORF1/VR1012, 14HWORF2/VR1012, 27HWORF1/VR1012, and HSP58/VR1012 were not protective and were excluded from the study. The remaining five DNA constructs-MAP2/ VR1012, 1HWORF3/ VR1012, 4HWORF1/ VR1012, 18HWORF1/ VR1012, and 3GDORF3/ VR1012-offered partial protection against lethal challenge demonstrated by reduced mortalities compared to control groups. Protection was augmented when DNA primed mice were boosted with a respective homologous recombinant protein. Protection in these five groups was associated with the induction of cell-mediated or T helper 1 (Th1) type of immune responses characterized by the production of large amounts of interferon gamma and interleukin-2 in in vitro proliferation assays using E. ruminantium antigens for stimulation. These responses were enhanced when the DNA-vaccinated DBA/2 mice were boosted with specific homologous recombinant protein vaccination. In a preliminary follow-up study, protection conferred by DNA vaccination with individual gene constructs was not enhanced when the protective constructs were administered in combination (including the map-1 gene of E. ruminantium). Further evaluation of these and other untested DNA constructs is necessary to optimize their expression in vivo in the presence of molecular adjuvants, such as the IFN gamma gene, GM-CSF gene, IL-12 gene, and CpG motifs to fully evaluate their protective value. PMID- 17114752 TI - New findings on members of the family Anaplasmataceae of veterinary importance. AB - Members of the family Anaplasmataceae are obligate intracellular Gram-negative bacteria that naturally infect a variety of wild and domestic animal species, the spillover of which may lead to zoonosis. I discuss new findings on members of the family Anaplasmataceae of veterinary importance and therefore, I will describe the recent findings on Neorickettsia risticii in the trematode and related Neorickettsia species. I also will review the recent progress on Aegyptianella pullorum and other Aegyptianella sp., "Candidatus Neoehrlichia mikurensis" and Anaplasma phagocytophilum strains in various hosts. The whole genome sequences of two important veterinary pathogens-Anaplasma marginale, the bovine anaplasmosis agent, and Ehrlichia (formerly Cowdria) ruminantium, the agent of heartwater of ruminants-have been published. Taken together, these advances in research of the family Anaplasmataceae in the veterinary field provide us with insights into the evolution, reservoir, and transmission of these organisms in nature and their pathogenesis in natural and accidental hosts. It is through this work that surveillance, diagnosis, preventive measures, and treatment of ehrlichioses of both animals and humans can be improved. PMID- 17114753 TI - Anaplasma phagocytophilum in ruminants in Europe. AB - The agent that causes tick-borne fever (TBF) in sheep was first described in 1940, 8 years after the disease was first recognized in Scotland. The same agent was soon shown to cause TBF in sheep and pasture fever in cattle in other parts of the UK, Scandinavia, and other parts of Europe. After the initial use of the name Rickettsia phagocytophila, the organism was given the name Cytoecetes phagocytophila to reflect its association with granulocytes and its morphological similarity with Cytoecetes microti. This name continued to be used by workers in the UK until the recent reclassification of the granulocytic ehrlichiae affecting ruminants, horses, and humans as variants of the same species, Anaplasma phagocytophilum. TBF and pasture fever are characterized by high fever, recurrent bacteremia, neutropenia, lymphocytopenia, thrombocytopenia, and general immunosuppression, resulting in more severe secondary infections such as tick pyemia, pneumonic pasteurellosis, listeriosis, and enterotoxemia. During the peak period of bacteremia as many as 90% of granulocytes may be infected. The agent is transmitted transtadially by the hard tick Ixodes ricinus, and possibly other ticks. After patent bacteremia, sheep, goats, and cattle become persistently infected "carriers," perhaps playing an important role in the maintenance of infection, in the flock/herd. Little is known about how efficiently ticks acquire and maintain infection in ruminant populations or whether "carrier" domestic ruminants play an important role as reservoirs of infection, but deer, other free living ruminants, and wild rodents are also potential sources of infection. During the late 1990s serological evidence of infection of humans was demonstrated in several European countries, creating a renewed interest and increased awareness of the zoonotic potential of TBF variants. More recently, a few cases of human granulocytic anaplasmosis (HGA) have been reported in some European countries, but it remains to be established whether the variants causing HGA in Europe are genetically and biologically different from those causing TBF in ruminants. TBF is readily diagnosed by demonstrating intracytoplasmic inclusions in peripheral blood granulocytes or monocytes of febrile animals or by detecting specific DNA by polymerase chain reaction (PCR), and TBF variants of A. phagocytophilum can be cultivated in tick cell lines, but the differentiation of TBF variants from HGA variants awaits further investigations. PMID- 17114754 TI - Epidemiological survey of Ehrlichia canis and related species infection in dogs in eastern Sudan. AB - The infection rates of Ehrlichia canis and related species in dogs in eastern Sudan were examined using molecular methods. Among 78 dogs examined, 63 (80.8%), 19 (24.4%), and 26 (33.3%) were positive for E. canis, Anaplasma platys, Mycoplasma haemocanis, and "Candidatus Mycoplasma haemoparvum," respectively. Among these, 30 dogs were single-positive: 25 for E. canis, 2 for A. platys, 1 for M. hemocanis, and 2 for "C. M. haemoparvum." The rest of the dogs (48.7%) were positive for two or more pathogens. PMID- 17114755 TI - Surveys on seroprevalence of canine monocytic ehrlichiosis among dogs living in the Ivory Coast and Gabon and evaluation of a quick commercial test kit dot ELISA. AB - Canine monocytic ehlichiosis (CME), an enzootic disease in Africa, has been studied in canine blood samples (serum). These dogs, without any clinical sign of disease, were living in Abidjan (Ivory Coast) and in several small villages located in northeasst Gabon (Ogooue Ivindo). The results obtained by indirect fluorescent antibody (IFA) test, used as a point of reference, and by a quick test dot-Elisa were compared. Blood samples taken from 390 asymptomatic dogs in 2003 (137 in Ivory Coast and 253 in Gabon) were screened by IFA (antigen from Symbiotics Europe, Lyon) with a positive threshold set at 1/80. Afterwards, CME was detected by the commercial test kit Dot-Elisa in solid phase Snap 3Dx (Idexx, Westbrook, Maine, USA), using recombinant proteins which belong to Ehrlichia canis, p30 and p30-1. Using the IFA test, CME seroprevalence in the Ivory Coast is found to be 67.8%. Among 93 Ivorian seropositive blood samples, 76 samples show an antibody titer >1/2560. In Gabon, IFA showed that seroprevalence is only 3.1%. Among 8 seropositive Gabonese dogs, only one sample shows an antibody test titer> 1/2560. Results from the Snap 3Dx test used on 390 blood samples are 100 positive samples and 290 negative ones. Comparison between IFA and Snap test 3DX revealed that the Snap test shows 97.9% specificity, 93.1% sensitivity, a 94% positive predictive value, a 97.6%, negative predictive value, and 96.6% reliability. In conclusion, CME seroprevalence in Abidjan is very high. Dogs studied for CME were watchdogs, living in kennels, where infection transmitted by Rhipicephalus sanguineus seems to be higher in the Gabonese area called Ogooue Ivindo, where semi-stray dogs were subjected to the test. These dogs where carried ticks identified as Haemaphysalis leachi, but this kind of tick is not considered as bearing Ehrlichia canis. Finally, results of the Snap 3Dx show that it is a simple and reliablemeans for quickly detecting dogs suspected of asymptomatic canine ehrlichiosis. PMID- 17114756 TI - Experimental Infections in dogs with Ehrlichia canis strain Borgo 89. AB - This study attemps to clarify the virulence and the pathogenicity of the Borgo 89 strain of Ehrlichia canis isolated from a sick dog in Corsica (France). Four unscathed beagles were intravenously injected with an inoculum of leukocytes infected with the Borgo 89 strain and the animals were examined daily for clinical signs of disease, and blood samples were drawn at frequent intervals for biochemical and hematologic assessment. Serologic (IFI) and PCR assays were also carried out. The results at autopsy are presented in this paper, leading to the conclusion that the Borgo 89 strain has a pathogenicity comparable to that of the known strains. However, the discovery of a case of completely unapparent infection raises the question of a possible individual immunization whose origin remains unexplained. PMID- 17114757 TI - Reservoir competency of goats for Anaplasma phagocytophilum. AB - The susceptibility of goats to infection by Anaplasma phagocytophilum (A. phagocytophilum) strains Ap-Variant 1 and Ap-ha was assessed by infestation of goats with field-collected Ixodes scapularis (I. scapularis) ticks. Both strains were infectious in the goat model. These results demonstrate that goats can be used in a laboratory setting to propagate A. phagocytophilum. Transmission from an infected goat to a naive goat by I. scapularis tick feeding was used to demonstrate that goats are reservoir-competent for the Ap-Variant 1 strain. PMID- 17114758 TI - An epidemiological study on Anaplasma infection in cattle, sheep, and goats in Mashhad Suburb, Khorasan Province, Iran. AB - The prevalence of Anaplasma infection was studied in cattle, sheep, and goats in the Mashhad area from 1999 to 2002. A total of 160 cattle from 32 farms and 391 sheep and 385 goats from 77 flocks were clinically examined for the presence of Anaplasma spp. in blood smears. The study revealed that 19.37% of cattle were infected with Anaplasma marginale and 80.3% of sheep and 38.92% of goats were infected with Anaplasma ovis. Prevalence of Anaplasma infection between male and female and between different age groups of cattle, sheep, and goats were statistically nonsignificant. Seasonally, the prevalence of Anaplasma infection in sheep and goats reached its highest level in summer, while a decrease was observed in autumn, and reached the lowest level in winter. The seasonal prevalence of Anaplasma infection in cattle was not significantly different. Symptomatic cases were not observed in any of the cattle, sheep, and goats. The ranges of anaplasmatemia in infected cattle, sheep, and goats were 0.005-0.5%, 0.01-3%, and 0.01-3%, respectively. PMID- 17114759 TI - Cytokine gene expression by peripheral blood leukocytes in dogs experimentally infected with a new virulent strain of Ehrlichia canis. AB - Ehrlichia canis (E. canis) is a lipopolysaccharide (LPS)-deficient obligatory intracellular bacterium that causes canine monocytic ehrlichiosis, a chronic febrile disease accompanied with hematological abnormality. This study analyzed temporal expression levels of IL-1beta, IL-2, IL-6, IL-8, IFN-gamma, and TNF alpha mRNA by peripheral blood leukocytes from dogs experimentally infected with a new virulent strain of E. canis by using real-time RT-PCR. Relative levels of IL-1beta and IL-8 transcripts normalized by the beta-actin transcript levels, were significantly upregulated, whereas those of TNF-alpha and IFN-gamma transcripts were only weakly upregulated in all three infected dogs, starting from 2 days up to 52 days post inoculation. The expressions of IL-2 and IL-6 genes were extremely low compared with the positive control (ConA-stimulated canine peripheral blood leukocytes). This study showed that E. canis can induce chronic expression of a subset of proinflammatory cytokine genes: balance, timing, and duration of these cytokine generations may contribute to the progression of canine ehrlichiosis. PMID- 17114760 TI - Serological evaluation of Anaplasma phagocytophilum infection in livestock in northwestern Spain. AB - A total of 1,098 serum samples were analyzed against Anaplasma phagocytophilum by immunofluorescent antibody (IFA) test. These serum samples belonged to four different populations distributed throughout two provinces of Galicia (Ourense and Pontevedra) located in northwestern Spain: bovine population (456 samples); ovine population (389 samples); caprine population (207 samples); and equine population (46 serum samples, all from Pontevedra). The seroprevalence against A. phagocytophilum within the bovine population was 3.07%. On the other hand, two of 389 (0.51%) sheep and one of 207 (0.48%) goats tested were seropositive, all of them showing low antibody titer. Seroprevalence within the equine population was 6.52% (3/46). Our results reveal the presence of antibodies against A. phagocytophilum in livestock from northwestern Spain, mainly in Pontevedra. PMID- 17114761 TI - Anaplasma phagocytophilum infection in cattle in France. AB - Anaplasma phagocytophilum is the agent of pasture fever or tick-borne fever, a disease of ruminants and humans in the United States and in Europe. Although several hundred cases have been suspected to occur in cattle in France, none has yet been microbiologically confirmed. We report the first identification of A. phagocytophilum 16S RNA gene sequence in a case of TBF in France. This indicates that the diagnosis of tick-borne fever should be also evoked in cattle exposed to Ixodes ticks in France. PMID- 17114762 TI - Understanding the mechanisms of transmission of Ehrlichia ruminantium and its influence on the structure of pathogen populations in the field. AB - The understanding of the structure of Ehrlichia ruminantium stock population in the field was highlighted by experiments done in controlled conditions on the goat model. The mixture of strains observed in ticks seemed to be due to simultaneous infections rather than successive infections of the carrier. During a dual infection, the timing of Ehrlichia ruminantium circulation of the two stocks in hosts influenced their selection by ticks. PMID- 17114763 TI - Incidence of ovine abortion by Coxiella burnetii in northern Spain. AB - The infectious causes of ovine abortion occurring in 148 farms in northern Spain between 1999 and 2003 were investigated. Laboratory analysis included microbiological, serological, pathological and molecular techniques. Border disease was diagnosed in 16% of the flocks, toxoplasmosis in 15%, chlamydiosis in 12%, salmonellosis in 10%, Q fever in 3%, miscellaneous infections in 7% (Yersinia spp., Listeria spp., Brucella spp.), and inflammatory lesions compatible with an infectious cause were seen in 7% of the flock. In an additional 1% of the flocks non-infectious causes were identified, and a diagnosis was not reached in 38% of the flocks. When a PCR retrospective study was carried out to investigate the possible implication of Coxiella burnetii in the cases without diagnosis, including those with inflammatory lesions, the prevalence of this pathogen increased from 3% up to 9% of the flocks, revealing the importance of this zoonotic pathogen as a small-ruminant abortifacient agent. Placenta was the most commonly positive sample, but other fetal tissues were also of value for C. burnetii DNA detection. The present results update information about the situation of abortion in sheep farms in northern Spain, and highlight the relevance of molecular diagnostic tools in routine laboratory analysis of abortions by C. burnetii. PMID- 17114764 TI - Detection of Coxiella burnetii in market chicken eggs and mayonnaise. AB - We tried to detect C. burnetii in market chicken eggs and mayonnaise by nested PCR assay. The PCR target was the com 1 gene of C. burnetii. The positive rate for egg and mayonnaise samples was 4.2% and 17.6%, respectively. Direct sequence of some of the positive egg samples shows mutations whereas no mutation was found in the positive mayonnaise samples. The number of molecules of the Q fever agent is estimated at 10(4) to 10(6) per egg, according to our quantitative PCR test. PMID- 17114765 TI - Efficacy of several anti-tick treatments to prevent the transmission of Rickettsia conorii under natural conditions. AB - The efficacy of several anti-tick treatments to prevent the transmission of Rickettsia was evaluated under natural conditions of tick pressure in a kennel. Only Amitraz (Preventic) provided total control on transmission (no dogs were infected), whereas with Frontline, Advantix, or Scalibor, the rate of infection varied among the dogs. PMID- 17114766 TI - Rickettsia spp. in Ixodes ricinus ticks in Bavaria, Germany. AB - This study aims to provide information on the occurrence of spotted fever rickettsiae in Ixodes ricinus ticks in southern Germany. A total of 2,141 I. ricinus ticks was collected in Bavaria. Pools of 5-10 ticks were studied by a PCR targeting the rickettsial citrate synthase gene gltA. The average prevalence rate was 12% (257 of 2,141). Sequencing data exclusively identified Rickettsia helvetica DNA. Results and other data demonstrate the possible role of R. helvetica in I. ricinus as a source of human infections in southern Germany. PMID- 17114767 TI - The occurrence of Spotted Fever Group (SFG) Rickettsiae in Ixodes ricinus ticks (Acari: Ixodidae) in northern Poland. AB - Ixodes ricinus, the most commonly observed tick species in Poland, is known vector of microorganisms pathogenic for humans as TBE virus, Borrelia burgdorferi s.1., Anaplasma phagocytophilum and Babesia sp. in this country. Our study aimed to find out whether this tick can also transmit also rickettsiae of the spotted fever group (SFG). DNA extracts from 560 ticks (28 females, 34 males, and 488 nymphs) collected in different wooded areas in northern Poland were examined by PCR for the detection of Rickettsia sp., using a primer set RpCS.877p and RpCS.1258n designated to amplify a 381-bp fragment of gltA gene. A total of 2.9% ticks was found to be positive. The percentage of infected females and males was comparable (10.5% and 11.8%, respectively) and 6.6-7.6 times higher than in nymphs (1.6%). Sequences of four PCR-derived DNA fragments (acc. no. DQ672603) demonstrated 99% similarity with the sequence of Rickettsia helvetica deposited in GenBank. The results obtained suggest the possible role of I. ricinus as a source of a microorganism, which recently has been identified as an agent of human rickettsioses in Europe. PMID- 17114768 TI - Molecular survey of Ehrlichia canis and Anaplasma phagocytophilum from blood of dogs in Italy. AB - We investigated the prevalence of Ehrlichia canis (E. canis) (n = 601) and Anaplasma phagocytophilum (A. phagocytophilum) (n = 460) infection by means of real-time PCR from blood of Italian dogs. The prevalence of E. canis in northern, central, and southern Italy was 2.9%, 8%, and 9.7%, respectively. The prevalence of A. phagocytophilum was 0%. PMID- 17114769 TI - Spotted fever group rickettsial infection in dogs from eastern Arizona: how long has it been there? AB - A serosurvey of free-roaming dogs for antibodies to spotted fever group rickettsiae was conducted using archival samples that had been collected in the White Mountain region of eastern Arizona during a plague study in 1996. Immunoglobulin G antibodies to Rickettsia rickettsii (5.1%) and to R. rhipicephali (3.6%) were demonstrated, and no cross-reactive samples were identified. This study indicates that R. rickettsii was likely present in the dog populations in this area prior to the recognition of human cases of Rocky Mountain spotted fever (RMSF). The role of dogs as short-term reservoirs and primary hosts for the vector tick, Rhipicephalus sanguineus, should receive closer attention. PMID- 17114770 TI - Isolation of Rickettsia rickettsii and Rickettsia bellii in cell culture from the tick Amblyomma aureolatum in Brazil. AB - Brazilian spotted fever (BSF) is a highly lethal disease caused by Rickettsia rickettsii. In the present study, rickettsial infection was evaluated in 669 Amblyomma aureolatum adult ticks collected from naturally infested dogs in Taiacupeba, a BSF-endemic area in the state of Sao Paulo. Ten (1.49%) ticks were infected with Rickettsia bellii, and 6 (0.89%) ticks were infected with R. rickettsii. Both Rickettsia species were isolated and established in Vero cell cultures. The Rickettsia isolates were characterized by molecular analyses, sequencing fragments of different rickettsial genes. Our results suggest that A. aureolatum is an important vector of R. rickettsii in Brazil. PMID- 17114771 TI - Multiplexed serology in atypical bacterial pneumonia. AB - Atypical pneumonia is a term applied to lower respiratory tract infections that are not characterized by signs and symptoms of lobar consolidation. This article will discuss the epidemiology, clinical manifestations, and laboratory diagnoses of Mycoplasma pneumoniae, Chlamydia sp., Legionella sp., Francisella tularensis, and Coxiella burnetii, which are the agents most commonly associated with atypical pneumonia. Because many of these pathogens are intracellular, diagnosis depends upon serological confirmation. The current serological tests used to identify these agents in the etiologic diagnosis of atypical pneumonia are described. Recently, however, it has become possible to make a diagnosis directly in these cases using DNA or protein microarrays. Here, we describe the development of a new, automated technique for simultaneous testing and detection of several pathogens using a multiplexed serology test. This should prove to be a valuable tool for the rapid determination of patient status, allowing effective and efficient postexposure prophylaxis and treatment. PMID- 17114772 TI - Isolation of Anaplasma phagocytophilum strain Ap-variant 1 in a tick-derived cell line. AB - Ten isolates of the Ap-Variant 1 strain of Anaplasma phagocytophilum were made in the Ixodes scapularis (I. scapularis)-derived cell line, ISE6. Two isolates were obtained from laboratory-infected goats and eight isolates were obtained from field-collected I. scapularis ticks. Each isolate showed 16S rRNA sequences identical to those as previously described for the Ap-Variant 1 strain. These are the first tissue culture isolates of the Ap-Variant 1 strain and will allow for further characterization of the biological and antigenic properties of this strain. PMID- 17114773 TI - Human anaplasmosis: the first Spanish case confirmed by PCR. AB - We report a case of human anaplasmosis (HA) fulfilling the confirmation criteria: epidemiologic data and clinical picture compatible with HA; presence of a morulae within polymorphonuclear leukocyte; and positive PCR assay for Anaplasma phagocytophilum: This case report shows the presence of HA in Spain. PMID- 17114774 TI - Two cases of human granulocytic ehrlichiosis in Sardinia, Italy confirmed by PCR. AB - In this work we report the first two cases of human granulocytic ehrlichiosis (HGE) in Sardinia. In early September 2004, a 69-year-old woman (patient 1) was admitted to the Infectious Diseases Institute of Sassari for rickettsiosis like syndrome: high fever (39.5-40 degrees C), dyspnea, reduced consciousness, vomiting, and cutaneous rash. In late September 2004, a 30-year-old man (patient 2) with high fever was admitted for an evident palmar and oral erythema, edema of the labium, very intense arthralgia, myalgia, and dyspnea. In these two hospitalized patients, the diagnosis was made through indirect IgM and IgG immunofluorescent technique and confirmed by the presence of the specific DNA in the leukocytes. The two patients were A. phagocytophilum-PCR positive. PMID- 17114775 TI - Multiplex detection of Ehrlichia and Anaplasma pathogens in vertebrate and tick hosts by real-time RT-PCR. AB - Tick-borne rickettsial infections are responsible for many emerging diseases in humans and several vertebrates. These include human infections with Ehrlichia chaffeensis, Ehrlichia ewingii and Anaplasma phagocytophilum. As single or co infections can result from a tick bite, the availability of a rapid, multiplex molecular test will be valuable for timely diagnosis and treatment. We recently described a muliplex-molecular test that can detect single or co-infections with up to five Ehrlichia and Anaplasma species. We reported that the test has the sensitivity to identify single infections in the canine host with E. chaffeensis, E. canis, E. ewingii, A. phagocytophilum, and A. platys and co-infection with E. canis and A. platys. In this study, ticks were collected from different parts of the state of Kansas during summer months of the year 2003 and tested for the presence of infection using the molecular test. The analysis revealed a minimum of 3.66% of the ticks to be positive for either E. chaffeensis or E. ewingii in A. americanum and Dermacenter species. This assay will be valuable in monitoring infections in dogs and ticks, and with minor modifications it can be used for diagnosing infections in people and other vertebrates. PMID- 17114776 TI - Identification and characterization of Coxiella burnetii strains and isolates using monoclonal antibodies. AB - We evaluated 6 monoclonal antibodies (MAbs) for their usefulness in identifying and characterizing recognized laboratory strains as well as field isolates of Coxiella burnetii. Five had been generated in response to strain Nine Mile (3 IgM class, 1 IgG class, 1 light chain producers only) and were polypeptide-specific, and 1 was anti-Priscilla (IgG class) and was lipopolysaccharide (LPS)-specific. Initially, the MAbs were used in conjunction with a dot blot assay with which we could differentiate C. burnetii from rickettsiae or chlamydiae. Confirmation of the specificity of these MAbs was provided by demonstrating that only C. burnetii antigens were recognized by certain combinations of antibodies used for immunoblotting proteins of various C. burnetii strains. Subsequently, we characterized antigens of 11 C. burnetii field isolates and 3 reference strains by Western blotting with individual MAbs. MAb 921 and 922 (IgG class), MAb 241, 242, 384, 386, 614 (IgM class), and 7A5, 7A1 (light chain) consistently recognized a protein. Staining intensity differed, depending on the strain tested, and there was variability in the size of the antigen immunoreactive with MAb 14H (IgG class, LPS-specific). The most reactive region was at about 249 kD. Variability of reactivities with field isolates was seen in both the distribution of individual bands and their intensities. We conclude that an extensive immunoblotting technique may be useful for C. burnetii strain differentiation and routine identification of C. burnetii can be accomplished using this MAb-based dot blot assay. PMID- 17114777 TI - Comparison of four commercially available assays for the detection of IgM phase II antibodies to Coxiella burnetii in the diagnosis of acute Q fever. AB - Four commercially available serological assays for the detection of IgM phase II antibodies in patients with acute Q fever infection were compared using a panel of 23 serum samples from patients with acute Q fever and 88 control sera from blood donors. PMID- 17114778 TI - Evaluation of a real-time PCR assay to detect Coxiella burnetii. AB - We evaluated real-time PCR assays for the detection of C. burnetii which targets sequences that are present either in one (icd) or in several copies (transposase of IS1111a) on the chromosome. The assays are highly sensitive, with reproducible detection limits of approximately 10 copies per reaction, at least 100 times more sensitive than capture ELISA, when performed on infected placenta material and specific for C. burnetii. The numbers of IS1111 elements in the genomes of 75 C. burnetii isolates were quantified by real-time PCR and proved to be highly variable. PMID- 17114779 TI - Diagnosis of acute Q fever by PCR on sera during a recent outbreak in rural south Australia. AB - Diagnosis of Q fever has largely been dependent upon serology, which may lead to delayed diagnosis as seroconversion can take weeks to develop. During a recent Q fever outbreak (27 patients) in rural South Australia, we compared the diagnostic rate of serology with two separate real-time PCRs, the 27kDa outer membrane protein and the insertion sequence. PCR was positive (on either or both PCR assays) in sera of 67% of the patients. Median time required for making serological diagnosis was 17 days, compared with 4 days by PCR. Q fever PCR is an effective tool in the diagnosis of acute Q fever infection. PMID- 17114780 TI - Evaluation of IgG antibody response against Rickettsia conorii and Rickettsia slovaca in patients with DEBONEL/TIBOLA. AB - The aim of the study was to determine the IgG antibody response to spotted fever group Rickettsia (SFGR) R. conorii and R. slovaca, and its specificity and sensitivity in patients with DEBONEL/TIBOLA. A prospective study of 31 patients with DEBONEL was carried out from January 2001 to May 2004. The SFGR serology testing (IgG IFA) for the diagnosis of DEBONEL/TIBOLA showed 61% sensitivity and 100% specificity. The R. slovaca antigen allowed the diagnoses in 18 of the 31 patients (58%), and 17 patients (55%) were diagnosed with this disease using R. conorii antigen. Therefore, using R. slovaca as antigen did not improve the sensitivity of the assay. PMID- 17114781 TI - Molecular typing of novel Rickettsia rickettsii isolates from Arizona. AB - Seven isolates of Rickettsia rickettsii were obtained from a skin biopsy, two whole-blood specimens, and from Rhipicephalus sanguineus ticks from eastern Arizona. Molecular typing of seven isolates of R. rickettsii and DNA samples from two other Rh. sanguineus ticks infected with R. rickettsii was conducted by PCR and DNA sequencing of rompA and 12 variable-number tandem repeat regions (VNTRs). All DNA specimens from Arizona were identical to each other and to reference human and Dermacentor andersoni isolates of R. rickettsii from Montana in their rOmpA gene sequences and 10 VNTRs. Two of the twelve VNTRs had differences in the number of repeat sequences in isolates from Arizona compared to those from Montana, thus conferring the novelty of the Rh. sanguineus-associated R. rickettsii. PMID- 17114782 TI - Ten years' experience of isolation of Rickettsia spp. from blood samples using the shell-vial cell culture assay. AB - Two strategies to improve the efficacy of the shell-vial method for Rickettsia were analyzed. Blood samples from 59 patients with Mediterranean spotted fever (MSF) were examined using the shell-vial technique. (i) DNA from positive lenses was obtained when they were contaminated. (ii) Blood sample from one patient was cultured in 17 shell-vials. R. conorii was identified in four cases by polymerase chain reaction (PCR)-RFLP. Three of these were obtained from cells adherent to lenses and the fourth one by using total patient blood sample. Rickettsia isolation using all blood samples as well as DNA from shell-vial lenses could be useful in the study of rickettsial infections. PMID- 17114783 TI - Automated method based in VNTR analysis for Rickettsiae genotyping. AB - A genetic locus named Rc-65, which is 5' adjacent to gene dksA and 3' adjacent to xerC gene, has previously been demonstrated to contain a VNTR with high discriminatory power in several rickettsial strains and thus, potentially useful for genetically similar strains identification. In this work, we present an automated molecular identification method based on capillary electrophoresis separation of VNTRs amplicons. The resulting electropherograms were in agreement with the sequence data obtained in a previous work. The presented genotyping method is fast and suitable for full automation, being a powerful tool for epidemiological surveillance in a large number of samples and enables the detection co-infected samples. The combination of other VNTR loci should improve the discriminatory capacity of this typing system, providing greater resolution and contributing to a more accurate VNTR-based assay. To our knowledge, this is the first automated assay for rickettsial strains identification. PMID- 17114784 TI - Monitoring of humans and animals for the presence of various Rickettsiae and Coxiella burnetii by serological methods. AB - Serological examination of humans in Slovakia suspected of having rickettsial infections revealed the presence of antibodies to spotted fever group rickettsiae (R. conorii, R. slovaca, and R. typhi). Of interest is the finding of serological positivity to the newly recognized "IRS" agent. Antibodies to these rickettsiae and to C. burnetii were demonstrated also in domestic and hunting dogs and pet animals. These results confirm the occurrence and possible circulation of these rickettsiae and C. burnetii in the Slovak Republic. PMID- 17114785 TI - Early diagnosis of rickettsioses by electrochemiluminscence. AB - The diagnosis of acute rickettsioses during the acute phase of the disease is challenging. We present preliminary evidence that antigen-capture using streptavidin-coated magnetic beads with biotinylated anti-rickettsia typhi rabbit polyclonal antibodies followed by electrochemiluminescent detection with ruthenylated antibodies of the same specificity could be used for the diagnosis of rickettsial diseases in the acute phase. PMID- 17114786 TI - Corpuscular antigenic microarray for the serodiagnosis of blood culture-negative endocarditis. AB - Blood culture-negative endocarditis is due to fastidious bacteria, including Coxiella burnetii and Bartonella spp. Diagnosis of such infection relies on serology and microimmunofluorescence is therefore the reference method. We developed a multiplex serology test featuring automatic incubation and reading and incorporating internal controls. Preliminary results indicate that this new serologic test is valuable for the rapid, automated serological diagnosis of blood culture-negative endocarditis. PMID- 17114787 TI - Proposal to create subspecies of Rickettsia sibirica and an emended description of Rickettsia sibirica. AB - The Rickettsia sibirica species is composed of isolates that are genotypically close but can be classified within two distinct serotypes, that is, R. sibirica sensu stricto and R. sibirica mongolitimonae (incorrectly named R. mongolotimonae). We investigated the possibility of classifying rickettsiae closely related to R. sibirica as R. sibirica subspecies, as proposed by the ad hoc Committee on Reconciliation of Approaches to Bacterial Systematics. For this, we first estimated the genotypic variability by using multilocus sequence typing (MLST), including the sequencing of five genes, and multispacer typing (MST) using three intergenic spacers, of five isolates and three tick amplicons of R. sibirica sensu stricto and six isolates of R. sibirica mongolotimonae. Then, we selected a representative of each MLST genotype and used mouse serotyping to estimate their degree of taxonomic relatedness. Among the 14 isolates or tick amplicons studied, 2 MLST genotypes were identified: (i) the R. sibirica sensu stricto type; and (ii) the R. sibirica mongolitimonae type. Representatives of the two MLST types were classified within three MST types and into two serotypes. Therefore, as isolates within the R. sibirica species are genotypically homogeneous but show MST genotypic, serotypic, and epidemio-clinical dissimilarities, we propose to modify the nomenclature of the R. sibirica species through the creation of subspecies. We propose the names R. sibirica subsp. sibirica subsp. nov. (type strain = 2-4-6, ATCC VR-541(T)), and R. sibirica subsp. mongolitimonae subsp. nov. (type strain = HA-91, ATCC VR-1526(T)). The description of R. sibirica is emended to accommodate the two subspecies. PMID- 17114788 TI - Comparison of immune response against Orientia tsutsugamushi, a causative agent of scrub typhus, in 4-week-old and 10-week-old scrub typhus-infected laboratory mice using enzyme-linked immunosorbent assay technique. AB - Using the indirect enzyme-linked immunosorbent assay (ELISA) technique, we compared the immune response of specific antibodies (IgM and IgG) in serum samples of scrub typhus-infected and non-infected mice against Orientia sutsugamushi (a causative agent of scrub typhus). Two different age groups (4 week-old and 10-week-old) of ICR laboratory mice were infected with O. tsutsugamushi by the animal passage procedure. Serum samples were detected for scrub typhus-specific antibodies using ELISA technique. Results from determining the presence of IgM and IgG in the serum samples obtained from scrub typhus infected mice showed that the IgG was first detected on day-13 after the initial infection in both the 4-week-old and 10-week-old mice. The IgG titer levels of both groups were not significantly different. Although the presence of IgM in the in serum samples obtained from scrub typhus-infected mice was first detected on day 13 in the 4-week-old mice and on day-12 in the 10-week-old mice, the IgM titer in the 4-week-old mice was slightly lower than in the 10-week-old mice. Clinical observations of the scrub typhus-infected mice showed that the older mice become ill on day 9 whereas the younger mice exhibited the symptoms on day 12. Considering the earlier detection and slightly higher level of specific IgM antibody, it could be interpreted that the older mice may appear to have responded against O. tsutsugamushi faster than the younger mice. PMID- 17114789 TI - Methods of isolation and cultivation of new Rickettsiae from the Nosoarea of the north Asian tick typhus in Siberia. PMID- 17114790 TI - Validation of a Rickettsia prowazekii-specific quantitative real-time PCR cassette and DNA extraction protocols using experimentally infected lice. AB - The presence of R. prowazekii in lice using R. prowazekii-specific qPCR cassettes showed the changes of the rickettsial burden in the lice post-infection. PMID- 17114791 TI - Identification of the hydrophobic ligand binding pocket of the S1P1 receptor. AB - Sphingosine 1-phosphate (S1P), a naturally occurring sphingolipid mediator and also a second messenger with growth factor-like actions in almost every cell type, is an endogenous ligand of five G protein-coupled receptors (GPCRs) in the endothelial differentiation gene family. The lack of GPCR crystal structures sets serious limitations to rational drug design and in silico searches for subtype selective ligands. Here we report on the experimental validation of a computational model of the ligand binding pocket of the S1P1 GPCR surrounding the aliphatic portion of S1P. The extensive mutagenesis-based validation confirmed 18 residues lining the hydrophobic ligand binding pocket, which, combined with the previously validated three head group-interacting residues, now complete the mapping of the S1P ligand recognition site. We identified six mutants (L3.43G/L3.44G, L3.43E/L3.44E, L5.52A, F5.48G, V6.40L, and F6.44G) that maintained wild type [32P]S1P binding with abolished ligand-dependent activation by S1P. These data suggest a role for these amino acids in the conformational transition of S1P1 to its activated state. Three aromatic mutations (F5.48Y, F6.44G, and W6.48A) result in differential activation, by S1P or SEW2871, indicating that structural differences between the two agonists can partially compensate for differences in the amino acid side chain. The now validated ligand binding pocket provided us with a pharmacophore model, which was used for in silico screening of the NCI, National Institutes of Health, Developmental Therapeutics chemical library, leading to the identification of two novel nonlipid agonists of S1P1. PMID- 17114792 TI - Control of adipose triglyceride lipase action by serine 517 of perilipin A globally regulates protein kinase A-stimulated lipolysis in adipocytes. AB - Phosphorylation of the lipid droplet-associated protein perilipin A (Peri A) mediates the actions of cyclic AMP-dependent protein kinase A (PKA) to stimulate triglyceride hydrolysis (lipolysis) in adipocytes. Studies addressing how Peri A PKA sites regulate adipocyte lipolysis have relied on non-adipocyte cell models, which express neither adipose triglyceride lipase (ATGL), the rate-limiting enzyme for triglyceride catabolism in mice, nor the "downstream" lipase, hormone sensitive lipase (HSL). ATGL and HSL are robustly expressed by adipocytes that we generated from murine embryonic fibroblasts of perilipin knock-out mice. Adenoviral expression of Peri A PKA site mutants in these cells reveals that mutation of serine 517 alone is sufficient to abrogate 95% of PKA (forskolin) stimulated fatty acid (FA) and glycerol release. Moreover, a "phosphomimetic" (aspartic acid) substitution at serine 517 enhances PKA-stimulated FA release over levels obtained with wild type Peri A. Studies with ATGL-and HSL-directed small hairpin RNAs demonstrate that 1) ATGL activity is required for all PKA stimulated FA and glycerol release in murine embryonic fibroblast adipocytes and 2) all PKA-stimulated FA release in the absence of HSL activity requires serine 517 phosphorylation. These results provide the first demonstration that Peri A regulates ATGL-dependent lipolysis and identify serine 517 as the Peri A PKA site essential for this regulation. The contributions of other PKA sites to PKA stimulated lipolysis are manifested only in the presence of phosphorylated or phosphomimetic serine 517. Thus, serine 517 is a novel "master regulator" of PKA stimulated adipocyte lipolysis. PMID- 17114793 TI - Rab GTPases containing a CAAX motif are processed post-geranylgeranylation by proteolysis and methylation. AB - Post-translational modification by protein prenylation is required for membrane targeting and biological function of monomeric GTPases. Ras and Rho proteins possess a C-terminal CAAX motif (C is cysteine, A is usually an aliphatic residue, and X is any amino acid), in which the cysteine is prenylated, followed by proteolytic cleavage of the AAX peptide and carboxyl methylation by the Rce1 CAAX protease and Icmt methyltransferase, respectively. Rab GTPases usually undergo double geranylgeranylation within CC or CXC motifs. However, very little is known about processing and membrane targeting of Rabs that naturally contain a CAAX motif. We show here that a variety of Rab-CAAX proteins undergo carboxyl methylation, both in vitro and in vivo, with one exception. Rab38(CAKS) is not methylated in vivo, presumably because of the inhibitory action of the lysine residue within the AAX motif for cleavage by Rce1. Unlike farnesylated Ras proteins, we observed no targeting defects of overexpressed Rab-CAAX proteins in cells deficient in Rce1 or Icmt, as reported for geranylgeranylated Rho proteins. However, endogenous geranylgeranylated non-methylated Rab-CAAX and Rab-CXC proteins were significantly redistributed to the cytosol at steady-state levels and redistribution correlates with higher affinity of RabGDI for non-methylated Rabs in Icmt-deficient cells. Our data suggest a role for methylation in Rab function by regulating the cycle of Rab membrane recruitment and retrieval. Our findings also imply that those Rabs that undergo post-prenylation processing follow an indirect targeting pathway requiring initial endoplasmic reticulum membrane association prior to specific organelle targeting. PMID- 17114794 TI - Mechanism of Dun1 activation by Rad53 phosphorylation in Saccharomyces cerevisiae. AB - Despite extensive studies, the molecular mechanism of DNA damage checkpoint activation remains incompletely understood. To better dissect this mechanism, we developed an activity-based assay for Dun1, a downstream DNA damage check-point kinase in yeast, using its physiological substrate Sml1. Using this assay, we confirmed the genetic basis of Dun1 activation. Rad53 was found to be directly responsible for Dun1 activation. We reconstituted the activation of Dun1 by Rad53 and found that phosphorylation of Thr-380 in the activation loop of Dun1 by Rad53 is responsible for Dun1 activation. Interestingly, phosphorylation of the evolutionarily conserved Thr-354 in the activation loop of Rad53 is also important for the regulation of Rad53 activity. Thus, this conserved mode of activation loop phosphorylation appears to be a general mechanism for the activation of Chk2 family kinases. PMID- 17114795 TI - Role of RAD51C and XRCC3 in genetic recombination and DNA repair. AB - In germ line cells, recombination is required for gene reassortment and proper chromosome segregation at meiosis, whereas in somatic cells it provides an important mechanism for the repair of DNA double-strand breaks. Five proteins (RAD51B, RAD51C, RAD51D, XRCC2, and XRCC3) that share homology with RAD51 recombinase and are known as the RAD51 paralogs are important for recombinational repair, as paralog-defective cell lines exhibit spontaneous chromosomal aberrations, defective DNA repair, and reduced gene targeting. The paralogs form two distinct protein complexes, RAD51B-RAD51C-RAD51D-XRCC2 and RAD51C-XRCC3, but their precise cellular roles remain unknown. Here, we show that, like MLH1, RAD51C localized to mouse meiotic chromosomes at pachytene/diplotene. Using immunoprecipitation and gel filtration analyses, we found that Holliday junction resolvase activity associated tightly and co-eluted with the 80-kDa RAD51C-XRCC3 complex. Taken together, these data indicate that the RAD51C-XRCC3-associated Holliday junction resolvase complex associates with crossovers and may play an essential role in the resolution of recombination intermediates prior to chromosome segregation. PMID- 17114796 TI - Cerebrospinal fluid dynamics between the intracranial and the subarachnoid space of the optic nerve. Is it always bidirectional? AB - CSF is thought to flow continuously from the site of production in the ventricles into interconnected spaces; i.e. cisterns and subarachnoid spaces (SASs). Since the SAS of the optic nerve is defined by a cul-de-sac anatomy, it is not evident how local CSF might recycle from that region to the general SAS. The concept of free communication of CSF has recently been challenged by the description of a concentration gradient of beta-trace protein, a lipocalin-like prostaglandin d synthase (L-PGDS), between the spinal CSF and that in the SAS of the optic nerve, indicating diminished local clearance or local overproduction of L-PGDS here. In fact, computed cisternography with a contrast agent in three patients with idiopathic intracranial hypertension and asymmetric papilloedema demonstrate a lack of contrast-loaded CSF in the SAS of the optic nerve despite it being present in the intracranial SAS, thus suggesting compartmentation of the SAS of the optic nerve. The concept of an optic nerve compartment syndrome is further supported by a concentration gradient of brain-derived L-PGDS between the spinal CSF and the CSF from the optic nerve SAS in the same patients. PMID- 17114797 TI - Prolonged activation of ASIC1a and the time window for neuroprotection in cerebral ischaemia. AB - Acid-sensing ion channels (ASICs), newly discovered members of epithelial Na+ channels/degenirin superfamily, are widely distributed throughout the mammalian peripheral and central nervous system and have been implicated in many physiological and pathophysiological processes. We have recently shown that activation of calcium-permeable ASIC1a is involved in acidosis-mediated, glutamate independent, ischaemic brain injury. In this study the neuroprotective time window for ASIC1a blockade in a mouse model of focal ischaemia is examined and the role of acidosis per se addressed by continuous pH measurements in penumbral cortex and post-ischaemic alkalization of brain. The effects of NMDA receptor blockade and ASIC1a blockade were compared. Specific ASIC1a blockade by the tarantula toxin psalmotoxin, PcTX, administered intracerebroventricularly as late as 5 h after 60 min of transient middle cerebral artery occlusion (MCAO) reduced infarct volume by >50%; the protection persisted for at least 7 days. Protection was also demonstrated after permanent MCAO. In penumbral cortex alkaline pH preceded acid pH and infarction. Attenuating brain acidosis by NaHCO3 or blocking ASIC1a with PcTX were both protective. NMDA blockade produced additive neuroprotection and the presence of PcTX prolonged the time window of effectiveness of NMDA blockade. Neuroprotection by PcTX was also achievable by intranasal administration. These findings further suggest that ASIC1a is a novel molecular target involved in ischaemic brain injury. Post-ischaemic administration of an ASIC1a blocker may prove to be an effective neuroprotective strategy for stroke patients. PMID- 17114798 TI - The link between mechanical stretch and glucose metabolism--a conceptual advance in understanding diabetic (and non diabetic?) renal disease. PMID- 17114799 TI - A painful hand in a kidney transplant recipient. PMID- 17114800 TI - What is a rheumatologist for? PMID- 17114801 TI - Influenza vaccination as model for testing immune modulation induced by anti-TNF and methotrexate therapy in rheumatoid arthritis patients. AB - OBJECTIVES: To compare serological response to influenza vaccine in patients with long-standing rheumatoid arthritis (RA) treated with tumour necrosis factor (TNF) blockers and/or methotrexate (MTX) and controls. METHODS: Altogether, 149 patients with RA and 18 healthy subjects were vaccinated. Fifty patients were treated with TNF blockers (etanercept or infliximab) in combination with MTX (TNF blockers + MTX), while 62 patients received TNF blockers alone or with other disease-modifying anti-rheumatic drugs (DMARDs) (TNF blockers without MTX). Thirty-seven patients were treated with MTX without TNF blockers (MTX). Vaccination was performed with trivalent vaccine (Influvax or Vaxigrip) both containing 15 microg haemagglutination inhibition (HI) of each of two A strains (H1N1 and H3N2) and one of B strains (B1 or B2). Serum samples were collected prior to and 4-6 weeks after vaccination and titrated against all four strains using HI assay. A positive immune response was defined as > or =4-fold increase compared with pre-vaccination titre levels. A titre > or =40 was considered protective. Pre- and post-vaccination geometric mean titres (GMT) were compared. RESULTS: Post-vaccination titre levels increased significantly in all groups, also reflected by high frequencies of positive immune responders. A positive immune response to combinations of all strains was significantly better for the MTX group. Individuals with protective levels before vaccination responded less well as a group. CONCLUSIONS: RA patients treated with MTX without TNF blockers had significantly better serological response to influenza vaccination compared with those receiving TNF blockers alone or in combination with MTX and/or other DMARDs. However, the immune response is sufficiently large to warrant influenza vaccination to all RA patients regardless of treatment. PMID- 17114802 TI - 18F-Flourodeoxyglucose positron emission tomography in polymyalgia rheumatica: novel insight into complex pathogenesis but questionable use in predicting relapses. PMID- 17114803 TI - Repetitive 18-fluorodeoxyglucose positron emission tomography in isolated polymyalgia rheumatica: a prospective study in 35 patients. AB - OBJECTIVE: To study fluorodeoxyglucose (FDG) deposition in different vascular beds and in the large joints of patients with isolated polymyalgia rheumatica (PMR), and to investigate whether there is a relation between FDG-positron emission tomography (PET) results and risk of relapse. METHODS: All consecutive patients with isolated PMR underwent a FDG-PET scan before treatment with steroids was started and--if logistics allowed--at 3 and 6 months. PET scans were scored at seven different vascular areas and a total vascular score (TVS) was calculated, ranging from 0 to 21. FDG uptake in the shoulders, the hips and the processi spinosi of the vertebrae was scored as 0 (no uptake), 1 (moderate uptake) or 2 (intense uptake). RESULTS: Thirty-five patients entered the study. At diagnosis, vascular FDG uptake was noted in 11 patients (31%), predominantly at the subclavian arteries. Mean TVS was low. FDG uptake in the shoulders was noted in 94% of patients, in the hips in 89% and in the processi spinosi of the vertebrae in 51%. The intensity of FDG uptake in the large vessels or in the shoulders, hips or processi spinosi did not correlate with the risk of relapse. CONCLUSIONS: Only one in three patients has an (moderately) increased vascular FDG uptake, especially in the subclavian arteries. The vast majority has inflammation of shoulders and hips, and half of them have increased FDG-uptake at the processi spinosi. Results of FDG-PET scans in patients with PMR did not correlate with their risk of relapse. PMID- 17114805 TI - Taking stock and moving forward. PMID- 17114804 TI - Histoplasmosis in a child with JRA on low-dose methotrexate. PMID- 17114806 TI - Regulation of intestinal NPC1L1 expression by dietary fish oil and docosahexaenoic acid. AB - To address the effect of the n-3 fatty acid, docosahexaenoic acid (22:6), on proteins that play a role in cholesterol absorption, CaCo-2 cells were incubated with taurocholate micelles alone or micelles containing 22:6 or oleic acid (18:1). Compared with controls or 18:1, 22:6 did not interfere with the cellular uptake of micellar cholesterol. Apical cholesterol efflux was enhanced in cells incubated with 22:6. Cholesterol trafficking from the plasma membrane to the endoplasmic reticulum was decreased by 22:6. 22:6 decreased Niemann-Pick C1-Like 1 (NPC1L1) protein and mRNA levels without altering gene or protein expression of ACAT2, annexin-2, caveolin-1, or ABCG8. Peroxisome proliferator-activated receptor delta (PPARdelta) activation decreased NPC1L1 mRNA levels and cholesterol trafficking to the endoplasmic reticulum, suggesting that 22:6 may act through PPARdelta. Compared with hamsters fed a control diet or olive oil (enriched 18:1), NPC1L1 mRNA levels were decreased in duodenum and jejunum of hamsters ingesting fish oil (enriched 22:6). In an intestinal cell, independent of changes in ABCG8 expression, 22:6 increases the apical efflux of cholesterol. 22:6 interferes with cholesterol trafficking to the endoplasmic reticulum by the suppression of NPC1L1, perhaps through the activation of PPARdelta. Moreover, a diet enriched in n-3 fatty acids decreases the gene expression of NPC1L1 in duodenum and jejunum of hamster. PMID- 17114807 TI - Chronic high-fat diet affects intestinal fat absorption and postprandial triglyceride levels in the mouse. AB - The effects of chronic fat overconsumption on intestinal physiology and lipid metabolism remain elusive. It is unknown whether a fat-mediated adaptation to lipid absorption takes place. To address this issue, mice fed a high-fat diet (40%, w/w) were refed or not a control diet (3%, w/w) for 3 additive weeks. Despite daily lipid intake 7.7-fold higher than in controls, fecal lipid output remained unchanged in mice fed the triglyceride (TG)-rich diet. In situ isolated jejunal loops revealed greater [1-(14)C]linoleic acid uptake without TG accumulation in mucosa, suggesting an increase in lipid absorption capacity. Induction both in intestinal mitotic index and in the expression of genes involved in fatty acid uptake, trafficking, and lipoprotein synthesis was found in high-fat diet mice. These changes were lipid-mediated, in that they were fully abolished in mice refed the control diet. A lipid load test performed in the presence or absence of the LPL inhibitor tyloxapol showed a sustained blood TG clearance in fat-fed mice likely attributable to intestinal modulation of LPL regulators (apolipoproteins C-II and C-III). These data demonstrate that a chronic high-fat diet greatly affects intestinal physiology and body lipid use in the mouse. PMID- 17114808 TI - Topology and membrane association of lecithin: retinol acyltransferase. AB - Fatty acid retinyl esters are the storage form of vitamin A (all-trans-retinol) and serve as metabolic intermediates in the formation of the visual chromophore 11-cis-retinal. Lecithin:retinol acyltransferase (LRAT), the main enzyme responsible for retinyl ester formation, acts by transferring an acyl group from the sn-1 position of phosphatidylcholine to retinol. To define the membrane association and localization of LRAT, we produced an LRAT-specific monoclonal antibody, which we used to study enzyme partition under different experimental conditions. Furthermore, we examined the membrane topology of LRAT through an N linked glycosylation scanning approach and protease protection assays. We show that LRAT is localized to the membrane of the endoplasmic reticulum (ER) and assumes a single membrane-spanning topology with an N-terminal cytoplasmic/C terminal luminal orientation. In eukaryotic cells, the C-terminal transmembrane domain is essential for the activity and ER membrane targeting of LRAT. In contrast, the N-terminal hydrophobic region is not required for ER membrane targeting or enzymatic activity, and its amino acid sequence is not conserved in other species examined. We present experimental evidence of the topology and subcellular localization of LRAT, a critical enzyme in vitamin A metabolism. PMID- 17114809 TI - Tumor necrosis factor-alpha-stimulated cell proliferation is mediated through sphingosine kinase-dependent Akt activation and cyclin D expression. AB - Tumor necrosis factor-alpha (TNF-alpha) has been shown to activate sphingosine kinase (SphK) in a variety of cell types. The extent to which SphK signaling mediates the pleiotropic effects of TNF-alpha is not entirely clear. The current study examined the role of SphK activity in TNF-alpha-stimulated cell proliferation in 1321N1 glioblastoma cells. We first demonstrated that pharmacological inhibitors of SphK markedly decrease TNF-alpha-stimulated DNA synthesis. Signaling mechanisms through which SphK mediated the effect of TNF alpha on DNA synthesis were then examined. Inhibition of Rho proteins with C3 exoenzyme or of Rho kinase with Y27632 attenuated TNF-alpha-stimulated DNA synthesis. However, RhoA activation by TNF-alpha was not blocked by SphK inhibition. ERK activation was also required for TNF-alpha-stimulated DNA synthesis but likewise TNF-alpha-induced ERK activation was not blocked by inhibition of SphK. Thus, neither RhoA nor ERK activation are the SphK-dependent transducers of TNF-alpha-induced proliferation. In contrast, TNF-alpha-stimulated Akt phosphorylation, which was also required for DNA synthesis, was attenuated by SphK inhibition or SphK1 knockdown by small interfering RNA. Furthermore, cyclin D expression was increased by TNF-alpha in a SphK- and Akt-dependent manner. Additional studies demonstrated that TNF-alpha effects on DNA synthesis, ERK, and Akt phosphorylation are not mediated through cell surface Gi -coupled S1P receptors, because none of these responses were inhibited by pertussis toxin. We conclude that SphK-dependent Akt activation plays a significant role in TNF-alpha induced cyclin D expression and cell proliferation. PMID- 17114810 TI - Comparison of angiotensin-converting enzyme, malonaldehyde, zinc, and copper levels in preeclampsia. AB - Preeclampsia is a syndrome of unknown etiopathogenesis. Recent studies carried out on preeclampsia have focused on the increase in free radicals in the feto placental unit with poor perfusion. It is believed that the renin-angiotensin system (RAS) has a role in the poor perfusion of the placenta. It is uncertain whether there is a pre-existing impairment in RAS in pre-eclamptic pregnant women or not. In the present study, we measured angiotensin-converting enzyme (ACE), malonaldehyde (MDA), zinc, and copper levels in the placental tissue of 16 pre eclamptic pregnant women and compared them with those in 20 healthy pregnant women. Whereas ACE activity and MDA were found to be high in the placentas of pre eclamptic patients, zinc and copper levels were low and there was a negative correlation between ACE activity and zinc concentration. These findings suggest that high ACE activity might play a role in the increase in tissue hypoxia and consequent lipid peroxidation through vasoconstriction; zinc deficiency in the placental tissue might cause insufficiency of superoxide dismutase, an antioxidant enzyme. Furthermore, deficiency in placental zinc also plays a role in the biosynthesis of connective tissue, maintaining its integrity, which might have an impact on the structure of the spiral arteries. PMID- 17114811 TI - Differentiation of serum levels of trace elements in normal and malignant breast patients. AB - The objective of this study was to determine which elements in serum best differentiated breast cancer in a case-control study. Concentrations of 13 elements in serum of 68 breast tumor patients (25 malignant and 43 benign) and 26 healthy controls were measured. Logistic regression with different variable selection procedures was used to determine a possible configuration of elements. Sensitivity and specificity of the model were calculated to obtain the optimal cutoff point for discriminating malignant breast cancers vs other individuals (including benign breast disease and normal ones). Acombination of Cd, Mn, and Fe was found to have a specificity and sensitivity of 100% using forward-type logistic regression, when the cutoff value of the combination score was 52.71. Using stepwise-type logistic regression, a combination of Cr and Mn had a sensitivity of 100% and a specificity of 97.1% when the combination score of 17.4 was chosen as the cutoff. Similar analysis could be implemented to compare the malignant and control groups. Specificity and sensitivity were 100% for Mn (forward and stepwise type) with a cutoff point of 6.40. For the backward regression, specificity was 84.6% and sensitivity was 100% for Zn, with a cutoff point of 869.1. In conclusion, there was a significant difference in concentrations of all 13 elements in serum between breast cancer patients and controls. A combination among Cd, Mn, Fe, Cr, and Zn might be important to determine a differentiating reference for breast cancers if a long-term followed up study is to be conducted. PMID- 17114812 TI - Essential trace and toxic element distribution in the scalp hair of Pakistani myocardial infarction patients and controls. AB - The pathogenesis of heart disease has been associated with changes in the balance of certain trace elements. The aim of this study was to evaluate the Zn, Fe, Cu, Cr, Ni, Pb, and Cd contents in scalp hair samples of myocardial infarction (MCI) patients hospitalized in the cardiac ward of National Hospital in Hyderabad city (Pakistan). Scalp hair samples were collected from 193 patients (104 male, 89 female) of 3 age groups (46-60, 61-75, and 76-90 yr), for a comparative study, 200 normal, healthy subjects (103 male, 97 female) of the same age groups residing in the same city were selected. All metals in scalp hair samples were assessed by a flame/ graphite furnace atomic absorption spectrophotometer, prior to microwave-assisted and conventional wet acid digestion methods. Results were calculated in micrograms per gram. The mean values of Fe and Zn of scalp hair samples of MCI patients were significantly reduced compared to the control subjects of both genders. The mean Fe concentrations in male patients were 19.42, 12.36, and 6.98 vs 30.69, 24.42, and 16.75 for the control patients in the three age groups (46-60, 61-75, and 76-90 yrs, respectively). The mean Zn concentration in male patients were 169.2, 149.4, and 107.7 microg/g vs 206.1, 188.0, and 154.4 microg/g for the control group (p < 0.002, 0.004, and 0.001) in all three age groups, respectively. These differences were also observed in the female study groups. The mean values of Pb, Cd, and Ni were significantly high in patients compared to healthy subjects (mean Pb in male patients: 11.85, 12.89, and 14.52 those of female patients were 11.88, 12.73, and 14.21 vs the male controls patients (6.08, 7.56, and 8.56) and female controls (5.99, 7.41, and 8.25) for all three age groups, respectively. The concentration of Ni and Cd in the scalp hair samples of the heart patients of both sexes were significantly higher compared to the control; in the case of Ni the range of significant difference for males was found to be p < 0.001-0.009 and for females to be p < 0.0.002-0.007 and significantly high concentration of Cd were observed in hair samples of patients than in controls in the range for males (p < 0.001-0.009) and in females (p < 0.001-0.011). The Zn/Cu and Zn/Cd ratios in the scalp hair (p < 0.01) of the diseased groups were significantly lower than that of the healthy groups. Deficiency of essential trace metals and high level of toxic metals might play a role in the development of heart disease in the subjects of this study. Toxic metals might also cause diminished absorption of essential elements. PMID- 17114813 TI - No meaningful increase in urinary tubular dysfunction markers in a population with 3 microg cadmium/g creatinine in urine. AB - The critical Cd exposure level to induce tubular dysfunctions is a focus of public concern among general populations in Japan. To answer this question, one group each (about 1000 adult women/area) in nonpolluted areas with high (Area H) and low Cd exposure (Area L) was obtained, and 742 strictly age-matched pairs of never-smoking adult women were selected for comparison. Cd, alpha1-MG (microglobulin) and beta2- MG in urine were taken as markers of exposure and tubular dysfunction, respectively. Geometric mean Cd levels as corrected for creatinine (Cdcr) was greater than three times higher in Area H (2.8 microg/g cr) than in Area L (0.8 microg/g cr). Nevertheless, beta2-MGcr did not differ between the two areas (125 microg/g cr for Area H vs 118 microg/g cr for Area L). alpha1 MGcr was only marginally higher in Area H (2.8 mg/g cr) than in Area L (2.1 mg/g cr), with no biomedical significance. Results were essentially the same when analyses were conducted with noncorrected observed values or values corrected for a specific gravity. Thus, the effects of Cd exposure in Area H on renal tubular function should be essentially nil. PMID- 17114814 TI - The effect of Zn on the Zn accumulation and biosynthesis of amino acids in mycelia of Cordyceps sinensis. AB - Zinc is an essential trace element in the human body and it participates in various pathways of metabolism. Cordyceps sinensis is a wellknown traditional Chinese medicine that contains cordycepin, cordycepic polysaccharides, proteins, vitamins, trace elements, and many other biological active materials. In this study, we cultured C. sinensis in liquid medium containing Zn ions and then analyzed the biomass, the ratios of total Zn accumulation, and the organic Zn content in the mycelia. The results showed that when the media contain 150 mg/L Zn, the biomass of mycelia of C. sinensis reached 10.7 g/L and the Zn content present in the mycelia reached 4875.1 microg/g. The percentage of Zn accumulated in the mycelia was 34.05% of the total Zn in the media, of which the organic Zn accounted for 76.33%. The results also revealed that the content of total amino acid (TAA) and essential amino acid (EAA) in the mycelia were increased substantially TAA and EAA in the Zn-treated mycelia were 11.1% and 15.2% higher, respectively, than that in the mycelia without Zn treatment. These results will be useful for elucidation of the physiological mechanism of Zn effects on the biological metabolism in the mycelia of C. sinensis. PMID- 17114815 TI - High-dose chromium(III) supplementation has no effects on body mass and composition while altering plasma hormone and triglycerides concentrations. AB - Chromium is generally believed to be an essential element and is often claimed to have value as a weight loss or muscle building agent. Recent studies in humans and rats have failed to demonstrate effects on body composition, although recent studies with pharmacological doses of the cation [Cr(III)3O(O2CCH2CH3)6(H2O)3]+ (or Cr3) (< or =1 mg Cr/kg body mass) in rats have noted a trend toward body mass loss and fat mass loss. Thus, the effects of large gavage doses of Cr3 (1-10 mg Cr/kg) on body mass, organ mass, food intake, and blood plasma variables (insulin, glucose, leptin, cholesterol, and triglycerides) were examined over a 10-wk period using male Sprague-Dawley rats. No effects on body composition were noted, although Cr3 administration lowered (p < 0.05) plasma insulin, leptin, and triglycerides concentrations. As Cr3 is absorbed greater than 10-fold better than commercially available nutritional supplements, the lack of an effect of the Cr(III) compound at these levels of administration clearly indicates that Cr(III) supplements do not have an effect on body composition at any reasonable dosage. PMID- 17114816 TI - Analysis of zinc and magnesium levels in pinealectomized chicks: roles on development of spinal deformity? AB - Melatonin is the main product of the pineal gland, and trace metals play a critical role in growth and development. The purpose of this study was to assess the serum zinc (Zn) and magnesium (Mg) levels in pinealectomized chicks and their possible interactions with the development of spinal deformity. Chicks were divided into two equal groups: unoperated controls (group M) and pinealectomized chicks (group N). Pinealectomies were performed at the age of 3 d. After 8 wk, serum Zn and Mg levels of 10 animals from each group were measured by spectrophotometric assay. The results of analyses were compared using the Mann Whitney U-test. The correlation between serum Zn and Mg levels were assessed by Spearman's correlation. In this study, it was obvious that the serum Zn levels in group N were significantly lower than those in group M (2.8 +/- 0.10 vs 4.2 +/- 0.14 ppm; p < 0.0005). In contrast, Mg levels in group N was high compared with the values in group M, although there was no significant difference (17.8 +/- 0.69 vs 15.7 +/- 0.85 ppm; p > 0.05). In pinealectomized animals, serum Zn levels declined significantly while serum Mg levels increased, albeit insignificantly. Thus, there was a moderately positive but not statistically significant correlation between Mg and Zn levels in unoperated controls (r = 0. 273, p > 0.05), whereas there was a negative but not statistically significant correlation between Zn and Mg levels in pinealectomized chicks (r = -0.115, p > 0.05). In addition, the serum Mg to serum Zn ratio was significantly higher in group N than in the group M control (6. 39 +/- 0.32 vs 3.75 +/- 0.22, respectively; p < 0.001). From the results of the current study, it is clear that surgical pinealectomy in newly hatched Hybro Broiler chicks has a significant effect on serum Zn level. However, the serum Mg did not change significantly. Because serum Mg is not a good indicator of Mg status in chicks, it is speculated that other tissues, such as muscle or spine, might productively be explored as a more sensitive Mg biomarker for this model. The present study provides experimental evidence that serum trace metal levels might be affected in pinealectomized animals because of the lack of its main neurohormone melatonin. PMID- 17114817 TI - Acorus calamus extracts and nickel chloride: prevention of oxidative damage and hyperproliferation response in rat kidney. AB - Nickel, a major environmental pollutant, is known for its clastogenic, toxic, and carcinogenic potential. In this article, we report the effect of Acorus calamus on nickel chloride (NiCl2)-induced renal oxidative stress, toxicity, and cell proliferation response in male Wistar rats. NiCl2 (250 micromol/kg body weight/mL) enhanced reduced renal glutathione content (GSH), glutathione- S transferase (GST), glutathione reductase (GR), lipid peroxidation (LPO), H2O2 generation, blood urea nitrogen (BUN), and serum creatinine with a concomitant decrease in the activity of glutathione peroxidase (GPx) (p < 0.001). NiCl2 administration also dose-dependently induced the renal ornithine decarboxylase (ODC) activity several-fold as compared to salinetreated control rats. Similarly, renal DNA synthesis, which is measured in terms of [3H] thymidine incorporation in DNA, was elevated following NiCl2 treatment. Prophylactic treatment of rats with A. calamus (100 and 200 mg/kg body weight po) daily for 1 wk resulted in the diminution of NiCl2- mediated damage, as evident from the downregulation of glutathione content, GST, GR, LPO, H2O2 generation, BUN, serum creatinine, DNA synthesis (p < 0.001), and ODC activity (p < 0.01) with concomitant restoration of GPx activity. These results clearly demonstrate the role of oxidative stress and its relation to renal disfunctioning and suggest a protective effect of A. calamus on NiCl2-induced nephrotoxicity in a rat experimental model. PMID- 17114818 TI - Upregulation of metallothionein-I mRNA expression in a rodent model for amyotrophic lateral sclerosis. AB - Metallothionein (MT) mRNA expression was investigated in a rodent model (G93A SOD1 transgenic mouse) for a lethal motor neuron disease, amyotrophic lateral sclerosis (ALS). In 8-wk-old mice that did not yet exhibit motor paralysis, MT-I mRNA expression was already significantly upregulated in the region of the spinal cord responsible for motor paralysis. The expression of another isoform, MT-III, was not changed. In the cerebellum, which is not responsible for motor paralysis in ALS, neither the expression profiles of MT-I nor MT-III were altered. In 16-wk old mice exhibiting motor paralysis, the expression of MT-I mRNA remained upregulated and the MT-III level tended to be elevated. Although no significant differences were found in the levels of both isoforms in the liver or kidney of 8 wk-old mice, the MT-I mRNA expression level was significantly upregulated in the kidney and liver of 16-wk-old mice. These results indicated that the MT-I isoform, but not the MT-III isoform, is associated with motor neuron death in ALS and suggested that the disease might be a systemic disorder to which the spinal cord is particularly susceptible. PMID- 17114819 TI - Effects of azadirachtin on six inorganic cation distributions in Ostrinia furnacalis (G.). AB - Azadirachtin (Az), as a botanical insecticide, is relatively safe and biodegradable. It affects a wide variety of biological processes, including the reduction of feeding, suspension of molting, death of larvae and pupae, and sterility of emerged adults in a dose-dependent manner. However,the mode of action of this toxin remains obscure. By using ion chromatography, we analyzed changes in six inorganic cation (Li+, Na+, NH4+, K+, Mg2+, and Ca2+) distributions of the whole body and hemolymph in Ostrinia furnacalis (G.) after exposure to sublethal doses of Az. The results showed that Az dramatically interfered with Na+, NH4 +, K+, Mg2+, and Ca2+ distributions in hemolymph of O. furnacalis (G.) and concentrations of these five cations dramatically increased. However, in the whole body, the levels of K+, Mg2+, and Ca2+ significantly, decreased after exposure to Az, except that Na+ and NH4 + remained constant. Li+ was undetected in both the control and treated groups in the whole body and hemolymph. It is suggested that Az exerts its insecticidal effects on O. furnacalis (G.) by interfering with the inorganic cation distributions related to ion channels. PMID- 17114820 TI - Autoimmune modulation of astrocyte-mediated homeostasis. AB - Astrocytes are principal mediators of homeostasis in the central nervous system (CNS). They supply neurons and oligodendrocytes with substrates for energy metabolism and clear the extracellular space of excess neurotransmitters. In neuroinflammation, astrocytes have classically been regarded as unimportant since their capacity to present antigen to T cells is limited and has been questioned in vivo. However, it is an evolving concept that autoimmunity in the CNS has a profound impact on astrocytes. In this review, we focus on the alterations in astrocyte functions that occur during an autoimmune attack of the CNS. PMID- 17114821 TI - Energy intake and amyotrophic lateral sclerosis. AB - Roy Walford, a physician and scientist who pioneered research on the anti-aging effects of caloric restriction and subjected himself to a low-energy diet, recently died from amyotrophic lateral sclerosis (ALS). Information from his case, epidemiological findings, and recent controlled studies in mouse models of ALS suggest that low-energy diets might render motor neurons vulnerable to degeneration, whereas high-energy diets are ameliorative. This contrasts with the effects of low-energy diets on various neuronal populations in the brain that respond adaptively, activating pathways that promote plasticity and resistance to disease. One reason that motor neurons might be selectively vulnerable to low energy diets is that they are unable to engage neuroprotective responses to energetic stress response involving the protein chaperones, such as, heat-shock protein-70. PMID- 17114822 TI - Pathomechanism of leukoaraiosis: a molecular bridge between the genetic, biochemical, and clinical processes (a mitochondrial hypothesis). AB - Ischemic demyelination in the white matter of the brain is a frequent clinical entity. In neuroimaging terms, it is referred to as leukoaraiosis (LA). LA can reflect a broad public health problem, which is caused by a cognitive impairment ranging from mild slowness of thinking to full-blown subcortical dementia. One quarter of subjects aged 65 yr or over are affected by some degree of white matter changes. There are a number of genetic factors that can be associated with circulatory disturbances of the white matter of the brain. A slight chronic hypoperfusion or an endothelial dysfunction associated with unfavorable genetic variations such as methylenetetrahydrofolate reductase C677T variation and angiotensin-converting enzyme I/D polymorphism then may lead indirectly to a malfunction of the molecular cross-talk between the nucleus and the mitochondria. This results in a decrease in the production of energy in the glia cells and thereby the beginning of demyelination. From another aspect, the presence of either the apolipoprotein E 2 or 4 alleles may cause an increased vulnerability to a slight chronic hypoperfusion of the white matter by reducing the range of mechanical and chemical flexibility of the glial cytoskeleton. In consequence of the chronic hypoperfusion, the functionally damaged kinesin protein gives rise also to the disturbances of the trafficking of the myelin basic protein mRNAs in the oligodendrocytes. On the basis of the current knowledge on LA, this article suggests a hypothetical molecular bridge between the genetic, biochemical, and clinical processes. PMID- 17114823 TI - Defects of immune regulation in the presenilin-1 mutant knockin mouse. AB - Mutations in the presenilin-1 (PS1) gene are causally linked to early-onset Alzheimer's disease (AD). Studies of neurons suggest that PS1 mutations result in a gain-of-function, which perturbs calcium regulation and increases cell vulnerability to apoptosis. Alterations in immune cell function have also been demonstrated in AD, and a role for PS1 in immune regulation has been suggested recently. We now report that splenocytes from PS1-mutant (M146V) knockin mice exhibit increased caspase activity, abnormal calcium regulation and aberrant mitochondrial function. Isolated splenic T cells from PS1-mutant mice respond poorly to proliferative signals and have downregulated cluster designation 3 and interleukin (IL)- 2-receptor expression necessary for a normal T-cell immune response. Thus, adverse effects of a mutation that causes AD on immune function that involves perturbed calcium regulation and cytokine signaling in lymphocytes, and associated sensitivity of lymphocytes to apoptosis are demonstrated. These findings suggest that abnormalities in immune function might play major roles in the pathogenesis of AD. PMID- 17114824 TI - Lack of minocycline efficiency in genetic models of Huntington's disease. AB - According to the recent controversy regarding the effects of minocycline in the R6/2 transgenic model of Huntington's disease (HD), this tetracycline has been re evaluated in another model, the N171-82Q strain. Ten miligrams per kilogram minocycline was given daily from the age of 2 mo, corresponding to an early symptomatic stage. We did not observe improvement in survival, weight loss, or motor function in treated transgenic mice. In addition, minocycline failed to mitigate the ventricle enlargement as well as the striatal and cortical atrophies induced by the transgene. Using high-performance liquid chromatography, it was observed that minocycline was similarly present in the plasma and the brain of both wild-type and N171-82Q mice following 14 daily injections. Using Western blot, we show that the increased expression of procaspase-1 induced by the transgene in the cortex was significantly reduced by the antibiotic. Combining together these data support that despite minocycline crosses blood-brain barrier in N171-82Q mice and displays an expected effect on procaspase-1 expression, it does not provide protection in this HD model. These in vivo results are in accordance with in vitro data, since minocycline failed to protect against mutated Huntingtin in an inducible PC12-clone expressing exon1 of mutated Huntingtin103Q. Altogether, the present data does not support minocycline as a beneficial drug for HD. PMID- 17114827 TI - Management of dyslipidemia and other cardiovascular risk factors in HIV-infected patients: case-based review. AB - Many HIV-infected patients have dyslipidemia and other cardiovascular risk factors prior to acquiring infection. Both HIV infection itself and antiretroviral therapy can cause or worsen lipid abnormalities. Management of dyslipidemia in the HIV-infected patient requires awareness of the effects of antiretroviral agents on lipid profiles, including potential sex- and race related effects, and interactions between lipid-modifying agents and antiretroviral agents. This article uses individual case histories to illustrate the decisions encountered in treating HIV infection and dyslipidemia. The article is based on a presentation on management of dyslipidemia and other cardiovascular risk factors in HIV infection made by Judith A. Aberg, MD, at the International AIDS Society-USA Los Angeles CME program in February 2006. PMID- 17114826 TI - Aluminum adjuvant linked to Gulf War illness induces motor neuron death in mice. AB - Gulf War illness (GWI) affects a significant percentage of veterans of the 1991 conflict, but its origin remains unknown. Associated with some cases of GWI are increased incidences of amyotrophic lateral sclerosis and other neurological disorders. Whereas many environmental factors have been linked to GWI, the role of the anthrax vaccine has come under increasing scrutiny. Among the vaccine's potentially toxic components are the adjuvants aluminum hydroxide and squalene. To examine whether these compounds might contribute to neuronal deficits associated with GWI, an animal model for examining the potential neurological impact of aluminum hydroxide, squalene, or aluminum hydroxide combined with squalene was developed. Young, male colony CD-1 mice were injected with the adjuvants at doses equivalent to those given to US military service personnel. All mice were subjected to a battery of motor and cognitive-behavioral tests over a 6-mo period postinjections. Following sacrifice, central nervous system tissues were examined using immunohistochemistry for evidence of inflammation and cell death. Behavioral testing showed motor deficits in the aluminum treatment group that expressed as a progressive decrease in strength measured by the wire-mesh hang test (final deficit at 24 wk; about 50%). Significant cognitive deficits in water-maze learning were observed in the combined aluminum and squalene group (4.3 errors per trial) compared with the controls (0.2 errors per trial) after 20 wk. Apoptotic neurons were identified in aluminum-injected animals that showed significantly increased activated caspase-3 labeling in lumbar spinal cord (255%) and primary motor cortex (192%) compared with the controls. Aluminum-treated groups also showed significant motor neuron loss (35%) and increased numbers of astrocytes (350%) in the lumbar spinal cord. The findings suggest a possible role for the aluminum adjuvant in some neurological features associated with GWI and possibly an additional role for the combination of adjuvants. PMID- 17114825 TI - Tryptamine induces tryptophanyl-tRNA synthetase-mediated neurodegeneration with neurofibrillary tangles in human cell and mouse models. AB - The neuropathological hallmarks of Alzheimer's disease (AD) and other taupathies include neurofibrillary tangles and plaques. Despite the fact that only 2-10% of AD cases are associated with genetic mutations, no nontransgenic or metabolic models have been generated to date. The findings of tryptophanyl-tRNA synthetase (TrpRS) in plaques of the AD brain were reported recently by the authors. Here it is shown that expression of cytoplasmic-TrpRS is inversely correlated with neurofibrillary degeneration, whereas a nonionic detergent-insoluble presumably aggregated TrpRS is simultaneously accumulated in human cells treated by tryptamine, a metabolic tryptophan analog that acts as a competitive inhibitor of TrpRS. TrpRSN- terminal peptide self-assembles in double-helical fibrils in vitro. Herein, tryptamine causes neuropathy characterized by motor and behavioral deficits, hippocampal neuronal loss, neurofibrillary tangles, amyloidosis, and glucose decrease in mice. Tryptamine induced the formation of helical fibrillary tangles in both hippocampal neurons and glia. Taken together with the authors' previous findings of tryptamine-induced nephrotoxicity and filamentous tangle formation in kidney cells, the authors' data indicates a general role of tryptamine in cell degeneration and loss. It is concluded that tryptamine as a component of a normal diet can induce neurodegeneration at the concentrations, which might be consumed along with food. Tryptophan-dependent tRNAtrp aminoacylation catalyzed by TrpRS can be inhibited by its substrate tryptophan at physiological concentrations was demonstrated. These findings indicate that the dietary supplementation with tryptophan as a tryptamine competitor may not counteract the deleterious influence of tryptamine. The pivotal role of TrpRS in protecting against neurodegeneration is suggested, providing an insight into the pathogenesis and a possible treatment of neurodegenerative diseases. PMID- 17114828 TI - Nucleoside analogue reverse transcriptase inhibitor options: a re-examination of the class. AB - The main options for dual nucleoside (or nucleotide) analogue reverse transcriptase inhibitors (nRTIs) as a component of initial antiretroviral therapy regimens are tenofovir/emtricitabine, zidovudine/lamivudine, and abacavir/lamivudine as fixed-dose combinations. Resistance to nRTIs can limit usefulness of many of the drugs in the class. Investigation of triple nRTI regimens has shown that zidovudine/lamivudine/abacavir does not provide benefits compared with dual nRTIs plus efavirenz and that others (tenofovir/lamivudine/abacavir and didanosine/lamivudine/abacavir) are associated with very high virologic failure rates. Further, 4-nRTI regimens are under investigation. The article summarizes a presentation on nRTIs made by Scott M. Hammer, MD, at the International AIDS Society-USA course in New York in March 2006. The original presentation is available as a Webcast at www.iasusa.org. PMID- 17114829 TI - Eye of the needle. PMID- 17114830 TI - Internet sex and dating sites need warnings. AB - For this issue's Commentary column, Peter L. Tenore, MD, discusses risk for infection with HIV or acquisition of sexually transmitted diseases associated with the use of the Internet to find sexual partners and presents results of an informal survey to determine whether Internet sites used to find sexual partners provide warnings about sexually transmitted diseases and HIV. He also calls for the operators of such sites to add warnings to their sites. PMID- 17114831 TI - Maternalizing the meninges: a pregnant Arabic legacy. PMID- 17114832 TI - Association between intravascular coagulopathy and outcome after traumatic brain injury. PMID- 17114833 TI - The coagulation system: a vulnerable system and a potential therapeutic aimpoint in treatment of traumatic brain injury. PMID- 17114834 TI - Genetic risk determinants in stroke: a global task. PMID- 17114835 TI - Recognition of impaired cognition in multiple sclerosis. PMID- 17114836 TI - Managing the first seizure: sharing the decision with the patient result in better care. PMID- 17114837 TI - Neuromyotonia: a diverse disorder. PMID- 17114838 TI - Mitochondrial complex-1 in Parkinson's disease. PMID- 17114839 TI - Inflammation in intracerebral hemorrhage: clearly present, but what is its role? PMID- 17114840 TI - Stereotaxy: the paradigm changes towards image-guided frameless procedures. PMID- 17114841 TI - Refractory status epilepticus. AB - Refractory status epilepticus (RSE) is a common problem in intensive care units and emergency departments. The important risk factor predisposing patients with SE to RSE is delay in receiving treatment. Self-sustaining SE is associated with progressive, time-dependent development of pharmacoresistance. Early termination of convulsive SE by aggressive treatment is the best way to prevent RSE. RSE once develop, requires more aggressive treatment as it is associated with higher mortality and morbidity. To date, no randomized controlled trials have been done for RSE. The most experience exists with coma inducing agents like pentobarbital, midazolam and propofol. New evidence suggests for the possible role of newer AEDs. PMID- 17114842 TI - Perspectives towards predictive testing in Huntington disease. AB - OBJECTIVE: Genetic counseling for individuals undergoing presymptomatic testing is lacking in India although testing is easily available. This has an impact on family members of Huntington's disease (HD), an autosomal dominant disease, wherein the age at onset of symptoms varies. AIM: We examine if attitudes differ towards presymptomatic testing for HD amongst HD family members, physicians and laypersons. MATERIALS AND METHODS: A modified questionnaire enquiring about opinions on various personal, family, social and future health care with regards to presymptomatic testing of HD was designed. A physician explained briefly about HD and presymptomatic testing of HD and recorded responses of unaffected family members of HD (n=25) and laypersons (n=50). Medical doctors (n=50) answered the questionnaire based on their knowledge of HD. RESULTS: HD family members, Medical doctors and laypersons were similar in their opinion to undergo the testing. Majority (60%) of HD family members did not wish to communicate test results with their friends when compared to the other two groups. Medical doctors and HD family members were more concerned about certainty of developing disease when the test results are positive. Majority (80%) of Medical doctors and less than half in the other groups felt that their decision to have a child would strongly depend on test results. Large proportion (80%) of HD family members did not wish to report their test results to their employers. CONCLUSIONS: Individuals with knowledge about HD and the test differ in their decision of sharing test results and reproductive choices. PMID- 17114843 TI - Fibrinolytic markers and neurologic outcome in traumatic brain injury. AB - AIMS: To determine the usefulness of fibrinolytic markers as early prognostic indicators in patients with isolated head trauma. MATERIALS AND METHODS: Sixty two consecutive patients (26 women and 36 men; mean age 61 years, range 2-76 years) with isolated head trauma seen within the first three hours of the trauma were included in the study. The Glasgow Coma score (GCS), platelet counts (Plt), prothrombin time (PT), partial thromboplastin time (PTT), fibrinogen, fibrin degradation products (FDP) and D-dimer levels were measured. Head computerized tomography (CT) findings were categorized as brain edema, linear fracture, depressed fracture, contusion and bleeding. Plt counts, PT, PTT, fibrinogen, FDP, D-dimer levels and CT findings were compared with both GCS and mortality in the first week. Statistical significance was accepted at P T) have been reported to affect the levels of IL-1 as well as its antagonist, IL-1Ra. It is also reported in several studies that these polymorphisms are associated with the susceptibility to cardio-cerebral vascular disease. However, data are limited in China. In this article, we studied the relationships between these polymorphisms and the risk of ischemic stroke in China. MATERIALS AND METHODS: One hundred and twelve patients committed ischemic stroke were compared with 95 demographically matched healthy volunteers. RESULTS: The frequencies of the IL-1Ra 1/1 genotype and IL-1Ra allele 1 (Ra*1 allele) in stroke patients were significantly higher than those in healthy volunteers [93.7% vs. 82.1%, P =0.014; 0.964 vs. 0.905, P =0.007]. No significant differences were found in the IL-1beta -511 genotype and the allele distribution between the two groups. CONCLUSIONS: Our results implicated that IL-1 gene polymorphism might be associated with the susceptibility to ischemic stroke. PMID- 17114845 TI - Differential patterns of memory performance in relapsing, remitting and secondary progressive multiple sclerosis. AB - BACKGROUND: Memory dysfunction is common in multiple sclerosis (MS). A retrieval failure has been reported as the primary cause for the memory deficits, although some studies also described a faulty acquisition. AIMS: The aim of the study was to examine memory function in relapsing remitting (RR) and secondary progressive (SP) MS patients, analyze the patterns of performance and to investigate whether disease course influences this performance. DESIGN AND SETTINGS: Case-control prospective study conducted in a clinical setting. MATERIALS AND METHODS: Fifty five RR, 23 SP MS patients and 80 normal subjects were evaluated with a comprehensive neuropsychological battery. Memory was assessed with tasks from the Signoret memory battery. Attention and executive function were also assessed. STATISTICAL ANALYSIS: Univariate analysis of variance, Mann-Whitney U-test, multivariate logistic regression and Chi-square test were used as appropriate. RESULTS: MS patients performed significantly worse than controls on almost all measures of memory (P < 0,001). MS subgroups differed in tasks of delayed recall (logical memory- P =0,019; wordlist delayed recall, P < 0,001), semantic cued recall (P < 0,001), recognition trials (P =0,006) rate of forgetting (P < 0,001) and confabulation and intrusion errors (P =0,004). CONCLUSIONS: Memory is consistently impaired in MS patients and disease course differentially affects the pattern of performance. SP patients show greater difficulties and a more pervasive pattern of dysfunction than RR patients. Delayed recall was the most affected memory measure and performance on this task discriminates between RR and SP MS patients. Relapsing remitting patients performed within the mildly impaired range while SP patients showed a moderate to severe impairment. PMID- 17114846 TI - Expanding traumatic intracerebral contusion/hematoma. AB - BACKGROUND AND AIMS: Delayed traumatic hematomas and expansion of already detected hematomas are not uncommon. Only few studies are available on risk factors of expanding hematomas. A prospective study was aimed to find out risk factors associated with such traumatic lesions. MATERIALS AND METHODS: Present study is based on 262 cases of intracerebral hematomas / contusions out of which 43 (16.4%) hematomas expanded in size. computerized tomography (CT) scan was done in all the patients at the time of admission and within 24 hours of injury. Repeat CT scan was done within 24 hours, 4 days and 7 days. Midline shift if any, prothrombin time, activated partial thromboplastin time, bleeding time, clotting time and platelet counts, Glasgow coma scale at admission and discharge and Glasgow outcome score at 6 months follow up were recorded. RESULTS: Twenty six percent, 11.3 and 0% patients developed expanding hematoma in Glasgow Coma scale (GCS) of 8 and below, 9-12 and 13-15 respectively. The chances of expanding hematomas were higher in patients with other associated hematomas (17.4%) as compared to isolated hematoma (4.8%) (Fisher's exact results P =0.216). All the cases of expanding hematoma had some degree of midline shift and considerably higher proportion had presence of coagulopathy. The results of logistic regression analysis showed GCS, midline shift and coagulopathy as significant predictors for the expanding hematoma. Thirty nine patients (90.7%) of the total expanding hematomas developed within 24 hours of injury. CONCLUSIONS: Enlargement of intracerebral hematomas is quite common and majority of them expand early after the injury. These lesions were common in patients with poor GCS, associated hematomas, associated coagulopathy and midline shift. PMID- 17114847 TI - Neuromyotonia: clinical profile of twenty cases from northwest India. AB - OBJECTIVES: We are presenting 20 cases of the intriguing clinico electromyographic entity, now considered a potassium channel disorder, Neuromyotonia. Our experience with the clinical manifestations, underlying abnormalities and response to various therapies is documented. MATERIALS AND METHODS: Patients with diffuse pain or undulating muscle movements, with or without stiffness were sent for electromyographic and further studies. Patients with "neuromyotonic discharges" were included after exclusion of hypocalcaemia. RESULTS: Our cases included 19 males and one female of age group 15 to 52 years, the majority being between 30 to 45 years. Undulating movements were seen in 19, of which two had focal twitching. Muscle stiffness was a complaint in five; pain was the chief presenting complaint of 19, which started in the calf in all. Irritability, insomnia and a peculiar worried pinched face were present in 12 patients. CSF was abnormal with mildly raised protein in eight. Curiously, 11 of these patients had taken ayurvedic treatment for various complaints in the preceding one month. Bell's palsy was associated in four, peripheral neuropathy in two and residual poliomyelitis in two. Electromyographic evidence of spontaneous activity in the form of "neuromyotonic discharges" was seen in all. Antibodies to voltage gated potassium channels was tested in one patient and was positive (titer was 1028 pM). Membrane stabilizers (e.g, phenytoin sodium) in our experience did not provide adequate rapid relief; we tried high-dose intravenous Methylprednisolone in 19 with significant amelioration of complaints. One patient was offered intravenous immunoglobulin, to which he responded. CONCLUSIONS: Neuromyotonia is a heterogeneous condition and can present in varied ways including diffuse nonspecific pain. This uncommon condition is potentially treatable and can be picked up with high index of suspicion. PMID- 17114848 TI - Patients' preferences towards antiepileptic drug therapy following first attack of seizure. AB - BACKGROUND: Antiepileptic drug (AED) therapy following first unprovoked seizure is controversial. AIM: To study the patients' preferences towards AED therapy following first unprovoked generalized tonic clonic seizure (GTCS). DESIGN: Prospective cohorts with one year follow-up study. SETTING: Government teaching hospital, a tertiary care center. MATERIALS AND METHODS: Patient cohort included patients with first attack of unprovoked GTCS within 30 days of onset, aged between 18-60 years and with normal brain CT scan. Counseling was done for all the patients and the relatives regarding seizure recurrence, duration and adverse effects of AED therapy if preferred. Patients were encouraged to make their own decision in preferring or deferring AED with reasons. They were followed up for one year. RESULTS: Of the 73 enrolled (54 males and 19 females) 39 (53%) preferred to go on AED therapy. The reasons for preferring AED therapy were; (a) fear of seizure recurrence, 21 (54%); (b) risky occupation, 14 (36%); and (c) fear of injury, 4 (10%). The reasons for deferring were: (a) fear of adverse effects of long-term AED therapy, 19 (56%) and (b) preferring to wait for the second attack, 15 (44%). All the patients were happy about being involved in the decision-making. CONCLUSION: Following first attack of unprovoked GTCS the decision regarding AED therapy may be taken by the patients and their family members after adequate counseling and such decisions have more relevance from their perspective. PMID- 17114849 TI - Respiratory-chain enzyme activities in isolated mitochondria of lymphocytes from patients with Parkinson's disease: preliminary study. AB - BACKGROUND: Evidence suggests that mitochondrial dysfunction stimulates the production of reactive oxygen species (ROS) that promote neural cell death in stroke and in Parkinson's disease. The sites of mitochondrial ROS production are not established but are generally believed to be located within the electron transport chain. AIMS: We studied the mitochondrial respiratory chain enzymes function from human circulating lymphocytes. SETTING AND DESIGN: Open study. MATERIALS AND METHODS: Forty patients with Parkinson's disease (PD) with 30 age matched control subjects were selected in this study. The patients had received no treatment before the study was conducted. STATISTICAL ANALYSIS: The data from patients and controls were compared using two-tailed student's t-test and values were expressed as means +/- standard deviation (SD). RESULTS: Respiratory complex I + III and IV activities were significantly lower (P < 0.001) in patients than in control subjects. CONCLUSIONS: The use of lymphocytes for investigating the respiratory chain enzymes provides an easy, noninvasive method to assess mitochondrial function in patients with PD. Furthermore, our study supports the hypothesis that a biochemical defect in the respiratory chain may be involved in the pathogenesis of PD. PMID- 17114850 TI - Comparative analysis of diagnostic accuracy of different brain biopsy procedures. AB - BACKGROUND: Image-guided procedures such as computed tomography (CT) guided, neuronavigator-guided and ultrasound-guided methods can assist neurosurgeons in localizing the intraparenchymal lesion of the brain. However, despite improvements in the imaging techniques, an accurate diagnosis of intrinsic lesion requires tissue sampling and histological verification. AIMS: The present study was carried out to examine the reliability of the diagnoses made on tumor sample obtained via different stereotactic and ultrasound-guided brain biopsy procedures. MATERIALS AND METHODS: A retrospective analysis was conducted of all brain biopsies (frame-based and frameless stereotactic and ultrasound-guided) performed in a single tertiary care neurosciences center between 1995 and 2005. The overall diagnostic accuracy achieved on histopathology and correlation with type of biopsy technique was evaluated. RESULTS: A total of 130 cases were included, which consisted of 82 males and 48 females. Age ranged from 4 to 75 years (mean age 39.5 years). Twenty per cent (27 patients) were in the pediatric age group, while 12% (16 patients) were >or= 60-years of age. A definitive histological diagnosis was established in 109 cases (diagnostic yield 80.2%), which encompassed 101 neoplastic and eight nonneoplastic lesions. Frame-based, frameless stereotactic and ultrasound-guided biopsies were done in 95, 15 and 20 patients respectively. Although the numbers of cases were small there was trend for better yield with frameless image-guided stereotactic biopsy and maximum diagnostic yield was obtained i.e, 87% (13/15) in comparison to conventional frame-based CT-guided stereotactic biopsy and ultrasound-guided biopsy. CONCLUSIONS: Overall, a trend of higher diagnostic yield was seen in cases with frameless image-guided stereotactic biopsy. Thus, this small series confirms that frameless neuronavigator-guided stereotactic procedures represent the lesion sufficiently in order to make histopathologic diagnosis. PMID- 17114851 TI - IgM anti-GM1 antibody titers in patients with monomelic amyotrophy. AB - BACKGROUND: Monomelic amyotrophy (MMA) is a benign motor neuron disorder, which particularly affects young people and the etiology is still unknown. Gangliosides are located on the outer surface of motor neurons. Anti-GM1 antibodies have been found to be elevated in multi-focal motor neuropathy with conduction block and other neurological diseases, which may have therapeutic implication. AIM: To evaluate IgM anti-GM1 antibody titers in patients of monomelic amyotrophy. SETTING AND DESIGN: prospective controlled study. MATERIALS AND METHODS: Forty six clinically and electrophysiologically diagnosed cases of MMA were assessed for IgM anti-GM1 antibody titers by enzyme-linked immunosorbent assay (ELISA) method and compared with titers in healthy controls, cases of amyotrophic lateral sclerosis (ALS) and acute inflammatory demyelinating polyneuropathy (AIDP). Titer of 800 units was taken as upper limit of normal (Buhlmann Laboratories AG, Switzerland). STATISTICAL ANALYSIS USED: one-way ANOVA. RESULTS: The mean age of 46 patients with MMA was 24.5 (+/- 7.3) years, with male female ratio of 44:2. The mean age of 19 healthy controls was 24.1 (+/- 3) years with male: female ratio of 18:1. Five (26%) individuals in the healthy control group, 22 (48%) patients of MMA, four (30%) of ALS and five (50%) of AIDP had high titers of IgM anti-GM1 antibody (P> 0.05). CONCLUSIONS: Although larger number of patients with MMA had higher IgM anti-GM1 antibody titers, the difference was not statistically significant from titers of healthy individuals and of patients in the ALS and AIDP group. PMID- 17114852 TI - Brain edema after intracerebral hemorrhage in rats: the role of inflammation. AB - BACKGROUND: Intracerebral hemorrhage (ICH) results in secondary brain edema and injury that may lead to death and disability. ICH also causes inflammation. It is unclear whether inflammation contributes to brain edema and neuron injury or functions in repairing the brain tissue. AIMS: To understand the effect of inflammation in ICH, we have carried out an investigation on the various aspects and the dynamic changes of inflammation. SETTINGS AND DESIGN: An ICH model was generated by injecting 50 microl autologous tail artery blood stereotactically into the right caudate nucleus of 30 rats, which were randomly divided into five ICH groups. Similarly, five Sham control groups were generated by inserting the needle to the right caudate nucleus of rats. MATERIALS AND METHODS: Rat behavior was evaluated over the time course (6 h, 24 h, 48 h, 72 h and 7 d) in each group. The rats were then killed by administering an overdose of pentobarbital. Following the euthanasia, the brain water content, neuronal loss, glia proliferation, inflammatory infiltration and brain morphology of the rats were measured. Additionally, the expression of TNF-alpha, IL-6, ICAM-1, VEGF, NF kappaB, C3 and CR2 was analyzed by immunohistochemistry. STATISTICAL ANALYSIS: The data were analyzed by student's t test. RESULTS: Rat brain water content increased progressively over the time course and reached its peak at 48 h followed ICH. The maximum of inflammatory infiltrate (especially neutrophils) and immunopositive cells of TNF-alpha, IL-6 and NF-kappaB, were at 48 h. The expression of C3 and CR2 reached their peaks at 48-72 h, while the expression ICAM-1 and VEGF were at maximum at 72 h followed ICH. CONCLUSIONS: The results suggested that the inflammatory cytokines, complement system and VEGF may have a function in the development of the brain edema and neuron injury followed ICH. PMID- 17114853 TI - Subclinical neurological involvement in Behcet's disease. AB - CONTEXT: Behcet's disease (BD) is a multisystem inflammatory disorder with unknown etiology characterized by recurrent oral and genital aphthous ulcers and uveitis. Behcet's disease can affect the central nervous system. AIMS: We aimed to investigate subclinical neurological involvement in patients who were suffering from BD and who had no neurological symptoms. SETTINGS AND DESIGN: A total of 49 patients were included in the study. For the investigation of subclinical neurological involvement, the patients received imaging and/or neurophysiologic evaluations. MATERIALS AND METHODS: The evaluation techniques were as follows: single photon emission computed tomography, 33 patients; cranial magnetic resonance imaging (MRI), 25 patients; brainstem auditory evoked potential examination, 36 patients; and electroencephalography (EEG), 30 patients. STATISTICAL ANALYSIS USED: The Mann-Whitney U test and Wilcoxon Rank Sum W test were used. RESULTS: Patients in the MRI and EEG groups showed significantly more abnormalities than did age- and gender-matched controls. CONCLUSIONS: Early diagnosis of neurological involvement in BD is important in reducing or preventing complications. Cranial MRI and EEG were found to be useful for detecting subclinical neurological abnormalities in patients with Behcet's disease. PMID- 17114854 TI - Ataxia and deafness in a young male: an unusual aetiology. AB - We report here a case of 18 year old male with tremors of hands, deafness, tendency to fall while walking, drowsiness and double vision of total duration 1(1/2) years. He had internuclear ophthalmoplegia, broken saccades, hypertonia and hyperreflexia of all four limbs, intention tremors, signs of gait and limb ataxia. Pupillary reactions and fundus examination were normal and signs of meningeal irritation or sensory neurological deficit were absent. MRI head and cervical spine with gadolinium enhancement revealed demyelination as evident from multiple oblong foci isointense on T1-weighted images and hyperintense on T2 weighted and fluid attenuated inversion recovery sequences in corpus callosum, sub-cortical white matter, right thalamus, pons and periaqueductal region of midbrain. Ill-defined linear hyperintense signals were observed in cervical spinal cord. No skeletal abnormality was noted in the skull or cervical spine. Oligoclonal bands were present in the cerebrospinal fluid. Brainstem auditory evoked potentials were abnormal, although visual evoked potentials were in normal range. A diagnosis of primary progressive multiple sclerosis (PPMS) was made fulfilling the revised criteria as laid down. In view of its presentation, it is a unique case of PPMS from India. PMID- 17114855 TI - Global aphasia due to left thalamic hemorrhage. AB - Global aphasia is an acquired language disorder characterized by severe impairments in all modalities of language. The specific sites of injury commonly include Wernike's and Broca's areas and result from large strokes--particularly those involving the internal carotid or middle cerebral arteries. Rarely, deep subcortical lesions may cause global aphasia. We present three cases with global aphasia due to a more rare cause: left thalamic hemorrhage. Their common feature was the large size of the hemorrhage and its extension to the third ventricule. HMPAO-SPECT in one of the cases revealed ipsilateral subcortical, frontotemporal cortical and right frontal cortical hypoperfusion. Left thalamic hemorrhage should be considered in the differential diagnosis of global aphasia. PMID- 17114856 TI - Sporadic onset Creutzfeldt-Jacob disease: interesting MRI observations. AB - We describe a 60-year-old woman with "probable" sporadic Creutzfeldt-Jacob disease (CJD) who manifested with two months history of rapidly progressive dementia and abnormal behavior, speech and gait abnormality, excessive sleepiness and myoclonic jerks. Scalp EEG showed diffuse slowing of background activity to delta range and triphasic sharp wave complexes occurring periodically twice in one-second interval. Magnetic resonance Imaging (MRI) of brain revealed high signal intensity on T2 weighted image (T2WI) and fluid attenuated inversion recovery sequences in the caudate and putamen bilaterally. Diffusion weighted images showed bilateral symmetric hyperintense signals in the caudate and putamen. The role of MRI in the diagnosis of CJD is discussed. PMID- 17114857 TI - Multiple simultaneous intracerebral hemorrhages following accidental massive lumbar cerebrospinal fluid drainage: case report and literature review. AB - Multiple simultaneous intracerebral hemorrhages (ICH) are uncommon. We report the case of an 80-year-old woman with previous diagnosis of normal pressure hydrocephalus and who was brought to our hospital with altered mental status and urinary incontinence. Medical history of hypertension, hematological disorders or severe head trauma was absent. Platelet count and coagulation profile were unremarkable. An initial head computed tomography (CT) showed sulcal enlargement and ventricular dilatation, but no evidence of ICH. A tap test indicated as a guide to case selection for shunt surgery accidentally resulted in cerebrospinal fluid (CSF) overdrainage. The patient presented sudden neurological deterioration, with sluggishly responsive pupils and generalized tonic-clonic seizures. A new head CT demonstrated multiple supra and infratentorial ICH. The patient became comatose and had a fatal course. Hence, CSF overdrainage may either cause or precipitate multiple simultaneous ICHs, affecting both the infratentorial and supratentorial regions. PMID- 17114858 TI - Transverse myelitis following spinal anesthesia. AB - Spinal anesthesia is widely used during surgical procedures. It is generally safe and the frequency of severe, permanent neurological complications associated with it has been reported to be extremely low. We report a patient, who developed paraplegia following spinal anesthesia. A 29-year-old male was referred with acute, flaccid, sensory motor paraplegia, with bladder and bowel involvement. He developed this immediately after an operation for inguinal hernia under spinal anesthesia. Spinal magnetic resonance imaging revealed hemorrhagic myelitis in the conus at D12. He was referred after he did not respond to intravenous methylprednisolone for 10 days. This case brings up the difficulty encountered in determination of the interspace used for spinal anesthesia and the potential for traumatic injury to the spinal cord. It also demonstrates the tragic outcome after a clinician violates some important, standard and established guidelines. PMID- 17114859 TI - Cases of glioblastoma multiforme metastasizing to spinal cord. AB - Cases of glioblastoma multiforme (GBM) metastasizing to the leptomeninx or the intramedullary spine are quite rare and prognoses are relatively poor. We present three cases of GBM with spinal metastasis, one of which also had leptomeningeal dissemination. Three patients with GBM were admitted to our clinic for postoperative radiotherapy after surgery. Leptomeningeal metastasis and dissemination were diagnosed with magnetic resonance imaging. Radiotherapy provided only temporary relief from pain with small improvement in neurological deficit but no survival advantage. PMID- 17114860 TI - Massive pontine hemorrhagic transformation associated with an anticoagulant for basilar artery occlusion. AB - Symptomatic hemorrhagic transformation is common in supratentorial and cerebellar infarction, but is rare in brainstem infarction. It is seldom reported in basilar artery occlusion. Although early arterial recanalization by thrombolytic agent has became the new trend of treatment, for some neurologists anticoagulant is still a conventional alternative treatment of basilar artery occlusion, especially in longer-existing ischemic deficits. We report a case of massive pontine hemorrhage associated with enoxaparin (low-molecular-weight heparin) treatment for basilar artery occlusion. On the basis of the clinical information and neuroimaging, an embolism was the most likely cause of stroke. The case presented herein adds massive pontine hemorrhagic transformation to the list of possible complications of anticoagulants for basilar artery occlusion. Apart from no evidence-based benefit in treatment of basilar artery occlusion, anticoagulant may contribute to devastating hemorrhagic transformation. PMID- 17114861 TI - Surgical management of a pial arteriovenous fistula with giant varix in an infant. AB - A seven-month-old infant presented with a generalized seizure. The radiological evaluation revealed a large arteriovenous fistula in the left sylvian region. The fistula connected the left middle cerebral artery and the vein of Trolard. A giant varix was present at the venous end. The child underwent craniotomy, complete disconnection of the fistula and excision of the varix. Except for recurrent seizures, which were eventually controlled on anticonvulsants, the child's neurological development has been good on long-term follow-up of three years. Pial arteriovenous fistulae are rare intracranial vascular malformations. Though significant operative risks exist, they can be successfully managed surgically with good long-term prognosis. PMID- 17114862 TI - Schwannoma of the oculomotor nerve. AB - A 63-year-old woman presented with an extremely rare oculomotor schwannoma not associated with neurofibromatosis, manifesting as a transient diplopia and ptosis. Magnetic resonance images showed a well-enhanced mass extending from the cavernous sinus to the intraorbital region. Surgical exposure confirmed the tumor originating from the oculomotor nerve in the cavernous sinus. The intraorbital cystic part of the tumor was partially resected to preserve the oculomotor function. This is the first case of oculomotor schwannoma in the cavernous sinus with intraorbital component. PMID- 17114863 TI - Diabetic nonketotic hyperosmolar state: interesting imaging observations in 2 patients with involuntary movements and seizures. AB - We report two patients of diabetic nonketotic hyperosmolar state presenting acutely with "self-limiting hemichorea - hemiballismus" and "generalized convulsive status epilepticus". CT scan in both the patients revealed a hyperdense nonenhancing basal ganglia. Magnetic resonance imaging brain of patient 1 showed it to be hyperintense on T1W image and iso-hyper intense on T2W image, minimally enhancing with contrast injection. PMID- 17114864 TI - Neurosurgical education. PMID- 17114865 TI - Stroke awareness program: the necessary facts for a successful campaign. PMID- 17114866 TI - Severe childhood autosomal recessive muscular dystrophy, mental subnormality and chorea. PMID- 17114867 TI - Cystathionine beta-synthase T833C/844ins68 polymorphism and stroke. PMID- 17114868 TI - Multiple dural sinus thrombosis: a differential diagnosis of a postpuncture headache in a puerperal patient. PMID- 17114869 TI - Incidentally detected intracranial sewing needles: an enigma. PMID- 17114870 TI - Ossification of the posterior longitudinal ligament of the thoracic spine in association with polycystic ovary syndrome. PMID- 17114871 TI - Adult onset hemiparkinsonism with brain hemihypoplasia. PMID- 17114872 TI - Rupture of a saccular microaneurysm of the supraclinoid internal carotid artery after mild head injury in a case with prominent posterior clinoid process. PMID- 17114873 TI - Supratentorial to infratentorial and antigravity migration of intracranial bullet. PMID- 17114874 TI - Double calvarial fibrous dysplasia. PMID- 17114875 TI - Smiling face odontoid. PMID- 17114876 TI - Ventilation and its control during incremental exercise in obesity. AB - BACKGROUND: In obesity, the addition of mass loading of the chest wall by adipose tissue decreases compliance, but its ventilation does not seem to be a limiting factor to physical performance. Plasma K(+) and lactic acid are considered important determinants of ventilation during exercise. Obesity is characterized by insulin resistance. OBJECTIVES: The aim of this study was to assess ventilatory adaptations to sustained effort and the effects of lactic acid and potassium in young obese subjects. METHODS: Twelve obese subjects with a body mass index of 40 (mean age 27 years, 6 males) and 12 normal subjects with a body mass index of 22 (aged 28 years, 6 males) performed a progressive cycloergometric test with increases of 20 W every 4 min to exhaustion while minute ventilation, oxygen consumption, carbon dioxide production, end-tidal oxygen pressure, and end tidal carbon dioxide pressure were measured. Blood samples were collected at the end of every step to determine plasma K(+). Lactic acid was measured at rest, 40, 80, 120 W and peak exercise (or only at peak exercise if <120 W). Before each exercise, we tested insulin sensitivity using the quantitative insulin sensitivity check index. RESULTS: Obese subjects had lower insulin sensitivity (0.318 vs. 0.345, p < 0.01). Peak exercise was not significantly different between both groups (125 W in the obese group vs. 137 W in the control group), but the ventilatory threshold was at lower power output in the obese group compared to the controls (76 vs. 107 W, p < 0.05). Ventilation increased less in the obese group but oxygen saturation of hemoglobin remained within normal limits up to exhaustion in both groups. Ventilation was appropriate for the CO(2) increase but less appropriate for the increased O(2) consumption. Both K(+) and lactic acid increased less in the obese group. CONCLUSIONS: In our obese subjects, ventilation was not a limiting factor during exercise. Its lower increase may be due, in addition to the characteristics of their chest walls, to insulin resistance which may limit the increase in lactic acid during effort, and to the hypertrophy of muscle fibers previously noted, which may be linked to a lower increase in plasma K(+) during physical exercise. PMID- 17114877 TI - Frequency and predictors of symptomatic intracerebral hemorrhage in patients with ischemic stroke treated with recombinant tissue plasminogen activator outside clinical trials. AB - BACKGROUND: To determine the frequency and predictors of symptomatic intracerebral hemorrhage (SICH) in patients treated with recombinant tissue plasminogen activator (rt-PA). METHODS: We reviewed the databases of 7 tertiary hospitals that treated ischemic stroke patients with intravenous rt-PA. We recorded demographic data, vascular risk factors, time between onset and treatment, dose, the NIHSS score, body temperature, blood pressure, platelet count, blood glucose, antiplatelet treatment, and CT data. We also registered the study protocol used for treatment and deviations from the accepted protocol. A control CT was performed on all patients. SICH was diagnosed if a parenchymal hematoma was detected within the 36 h after rt-PA and was associated with an increase of > or =4 in the NIHSS score. Bivariate analyses were performed followed by a logistic regression analysis. RESULTS: A total of 347 patients were studied, whose mean age was 68 +/- 10.9 years; 56% were men. Thirty-two patients (9.2%) exhibited a parenchymal hematoma, and 8 patients (2.3%) suffered a SICH. Patients with SICH had a higher frequency of previous transient ischemic attack (p = 0.04), early signs of ischemia (p = 0.003), hyperdense arterial sign (p = 0.008), and deviations (p = 0.002). Early signs of ischemia (OR 8.5, 95% CI 1.6 45.4, p = 0.01) and deviation from the protocol (OR 11.1, 95% CI 2.4-50, p = 0.002) were independent predictors of SICH. CONCLUSIONS: SICH is infrequent in patients with ischemic stroke treated with rt-PA outside of a clinical trial. Its frequency increases in the presence of early signs of ischemia on the non contrast CT scan and deviations from the recommended protocol. PMID- 17114878 TI - Revascularization compared to medical treatment in patients with silent vs. symptomatic residual ischemia after thrombolyzed myocardial infarction--the DANAMI study. AB - AIMS: The aim was to compare the effect of revascularization to conservative treatment in patients with residual silent and with residual symptomatic ischemia following acute myocardial infarction (AMI). The study was a subanalysis of the DANAMI (DANish AMI) randomized study of invasive vs. conservative treatment in patients with inducible ischemia after thrombolysis in AMI. METHODS AND RESULTS: One thousand and eight patients were randomized to invasive or conservative treatment, stratified by the type of ischemia: silent, i.e. ST depression during an exercise test prior to discharge in 56%, or symptomatic, i.e. chest pain occurring either spontaneously during admission or during the exercise test, with or without ST changes, in 44%. Compared to a conservative strategy, invasive treatment reduced the incidence of nonfatal reinfarction, after in median 2.4 years, in both symptomatic patients (13.3-7.2%, p < 0.006) and patients with silent ischemia (10.1 vs. 5.7%, p < 0.05), and of admissions with unstable angina in symptomatic (44.5-27.6%, p < 0.0001) and silent ischemia (21.6-13.3%, p < 0.0006). CONCLUSIONS: Compared to conservative strategy, invasive treatment reduces the risk of nonfatal reinfarction and hospital admissions for unstable angina in thrombolyzed post-AMI patients with silent as well as symptomatic exercise-induced ischemia. PMID- 17114879 TI - Exercise testing in asymptomatic aortic stenosis. AB - AIMS: To determine the safety of exercise testing (ET) in patients with moderate or severe asymptomatic aortic stenosis (AAS) and its accuracy to predict the need for surgery and mortality. METHODS: 106 consecutive patients with AAS performed a maximal ET. RESULTS: Follow-up [10.7 (4.9-19.4) months (percentile 25-75)] was completed in 102 patients (96.2%), 63.9 years (+/-15.1), 65 (61.3%) male, peak gradient 82.8 mm Hg (+/-25.4), mean gradient 50.5 mm Hg (+/-16.6), valve area 0.67 cm(2) (+/-0.16); 67 patients (65.7%) had abnormal ET. Among the 35 patients with normal ET, there were no deaths and 10 aortic valve replacements (AVR) (28.5%) were performed. Among the 67 patients with abnormal ET, 37 (55.2%) had events (35 AVR and 2 died) (p <0.0001). There were no complications with ET. CONCLUSION: ET may be performed safely in patients with AAS. ET gives additional information to an AVR decision. PMID- 17114880 TI - Efficacy and safety of clopidogrel 600 mg administered pre-hospitally to improve primary percutaneous coronary intervention in patients with acute myocardial infarction (CIPAMI): study rationale and design. AB - Clopidogrel, in combination with acetylsalicylic acid, has become a mainstay of the pharmacological therapy for patients with acute coronary syndromes, especially in those undergoing percutaneous coronary interventions (PCI). While a series of studies has shown that pre-treatment with a loading dose of clopidogrel 300 or 600 mg prior to PCI is effective in reducing cardiovascular complications, the optimal dose and timing in various patient groups is still unclear. The primary objective of the present randomized, open-label Clopidogrel to Improve Primary percutaneous coronary Intervention in Acute Myocardial Infarction (CIPAMI) study is to evaluate the efficacy and the safety of a 600 mg loading dose of clopidogrel in addition to standard acetylsalicylic acid/heparin treatment in the pre-hospital setting in 654 patients with acute ST elevation myocardial infarction scheduled for primary PCI. The primary efficacy endpoint is the TIMI 2/3 patency of the infarct-related artery immediately prior to PCI. The rationale, design and methods of this study are described. PMID- 17114881 TI - A current problem in cardiology: very late thrombosis after implantation of sirolimus eluting stent. AB - Discontinuation of antiplatelet medications has been strongly associated with coronary stent thrombosis. The first reported cases have been documented at 6 h to 6 weeks after stent implantation. This article presents a case of very late stent thrombosis 24 months after sirolimus eluting stent implantation and 18 months after clopidogrel discontinuation, despite aspirin continuation, and argues in favor of prolonging dual antiplatelet medication including clopidogrel in this setting, at least until data from randomized trials address this important issue. PMID- 17114882 TI - Depression in vascular dementia is quantitatively and qualitatively different from depression in Alzheimer's disease. AB - BACKGROUND/AIMS: To compare the prevalence and characteristics of depression in vascular dementia (VaD) and Alzheimer's disease (AD) after adjusting for dementia severity and gender. METHODS: One hundred and eight pairs of VaD and AD patients matched for dementia severity and gender were assessed. RESULTS: Major depressive disorder (MDD) was more prevalent in the VaD group than in the AD group (20.4% in VaD, 10.2% in AD, p = 0.04, Cochran-Mantel-Haenszel, CMH, test) regardless of the dementia severity and gender. The odds ratio for developing MDD in the VaD group versus the AD group was estimated to be 2.20 (95% confidence interval = 1.02 4.74). Neurovegetative symptoms such as 'felt tired and weak all the time' (30.6% in VaD, 13.9% in AD, p = 0.003, CMH test) and 'changed weight without trying' (16.7% in VaD, 6.5% in AD, p = 0.02, CMH test) were more prevalent in the VaD group than in the AD group. CONCLUSION: Depression in VaD was quantitatively and qualitatively different from that in AD regardless of the severity of dementia and gender; depression was more prevalent, severer and more retarded and vegetative in VaD than in AD. PMID- 17114883 TI - Preliminary assessment of cognitive function in older adults by clock drawing, box copying and narrative writing. AB - OBJECTIVE: To examine 3 cognitive screening tasks in older adults and determine which demographic, medical and lifestyle variables obtained 13-17 years previously predicted cognitive performance. DESIGN: Population-based longitudinal cohort study begun in 1981. SETTING: Leisure World Laguna Hills, a California retirement community. PARTICIPANTS: The cohort is predominantly white, well educated, upper-middle class; two thirds are women. Data from 1,744 participants (mean age 83 years, range 57-103) were analyzed. MEASUREMENTS: The baseline postal survey (1981-1985) asked demographic information, medical history, selected drug use and personal habits. The 1998 follow-up included 3 tasks to assess cognitive function: (1) clock drawing, (2) copying a box drawing and (3) narrative writing to describe a pictured scene. RESULTS: Within age groups, women tended to have lower scores than men on box copying, similar scores on clock drawing, but higher informational and complexity scores on narrative writing. Performance decreased with increasing age and was poorest in individuals > or = 90 years old. In addition to age, sex and education, significant predictors of performance were: exercise, body mass index and cataract surgery (clock); vitamin E supplements, daily aspirin use and gallbladder surgery (box); vitamin A supplements (narrative writing). Mean scores on all tasks were lower in those later identified as demented compared with nondemented but significantly different only for clock and box drawings. CONCLUSION: Performance on 3 cognitive tests showed significant effects of age, sex and education and was related to later dementia. Better performance by users of antioxidant supplements and aspirin suggests that these drugs may maintain cognitive function in later life. PMID- 17114884 TI - Severe bradycardia in a stroke patient caused by a single low dose of escitalopram. PMID- 17114885 TI - Transcortical sensory aphasia as a result of left frontal cortical-subcortical infarction. A case report. PMID- 17114886 TI - Backward genotype-trait association (BGTA)-based dissection of complex traits in case-control designs. AB - BACKGROUND: The studies of complex traits project new challenges to current methods that evaluate association between genotypes and a specific trait. Consideration of possible interactions among loci leads to overwhelming dimensions that cannot be handled using current statistical methods. METHODS: In this article, we evaluate a multi-marker screening algorithm--the backward genotype-trait association (BGTA) algorithm for case-control designs, which uses unphased multi-locus genotypes. BGTA carries out a global investigation on a candidate marker set and automatically screens out markers carrying diminutive amounts of information regarding the trait in question. To address the 'too many possible genotypes, too few informative chromosomes' dilemma of a genomic-scale study that consists of hundreds to thousands of markers, we further investigate a BGTA-based marker selection procedure, in which the screening algorithm is repeated on a large number of random marker subsets. Results of these screenings are then aggregated into counts that the markers are retained by the BGTA algorithm. Markers with exceptional high counts of returns are selected for further analysis. RESULTS AND CONCLUSION: Evaluated using simulations under several disease models, the proposed methods prove to be more powerful in dealing with epistatic traits. We also demonstrate the proposed methods through an application to a study on the inflammatory bowel disease. PMID- 17114887 TI - Rapid genetic analysis in congenital hyperinsulinism. AB - BACKGROUND: In severe, medically unresponsive congenital hyperinsulinism (CHI), the histological differentiation of focal versus diffuse disease is vital, since the surgical management is completely different. Genetic analysis may help in the differential diagnosis, as focal CHI is associated with a paternal germline ABCC8 or KCNJ11 mutation and a focal loss of maternal chromosome 11p15, whereas a maternal mutation, or homozygous/compound heterozygous ABCC8 and KCNJ11 mutations predict diffuse-type disease. However, genotyping usually takes too long to be helpful in the absence of a founder mutation. METHODS: In 4 patients, a rapid genetic analysis of the ABBC8 and KCNJ11 genes was performed within 2 weeks on request prior to the decision of pancreatic surgery. RESULTS: Two patients had no mutations, rendering the genetic analysis non-informative. Peroperative multiple biopsies showed diffuse disease. One patient had a paternal KCNJ11 mutation and focal disease confirmed by positron emission tomography scan and biopsies. One patient had a de novo heterozygous ABBC8 mutation and unexplained diffuse disease confirmed by positron emission tomography scan and biopsies. CONCLUSION: A rapid analysis of the entire ABBC8 and KCNJ11 genes should not stand alone in the preoperative assessment of patients with CHI, except for the case of maternal, or homozygous/compound heterozygous disease-causing mutations. PMID- 17114888 TI - Acylated ghrelin secretion is acutely suppressed by oral glucose load or insulin induced hypoglycemia independently of basal growth hormone secretion in humans. AB - BACKGROUND: Ghrelin has been reported to be the natural ligand of growth hormone (GH) secretagogue receptor, and it is known that exogenous ghrelin administration strongly stimulates GH release in humans. However, the effects of endogenous ghrelin on GH secretion and changes in ghrelin levels during dynamic changes in GH levels are not well understood. METHODS: Therefore, we measured circulating acylated ghrelin concentrations during oral glucose tolerance tests (OGTTs) in patients with active acromegaly (AA, n = 9) and in age/sex/BMI-matched group A controls (n = 12), and during insulin tolerance testing (ITT) in patients with GH deficiency (GHD, n = 10) and in group B controls (n = 10). Plasma acylated ghrelin, serum GH, insulin and glucose levels were measured during each test. RESULTS: Fasting plasma ghrelin levels correlated negatively with serum insulin levels in both group A and B controls (r = -0.665; p < 0.05) but not in patients with AA or GHD. During OGTTs, circulating ghrelin levels decreased significantly with a nadir at 30 min in both patients with AA (p < 0.05) and group A controls (p < 0.01). Also, ITTs were followed by a significant decrease in circulating ghrelin levels with a nadir at 30 min in patients with GHD (p < 0.05) and in group B controls (p < 0.05). CONCLUSION: The results of the study show that at baseline acylated ghrelin levels do not differ with respect to the GH status (GH excess or GH deficiency) and, furthermore, the suppression of acylated ghrelin levels during OGTT or ITT is independent of the GH response to the tests. PMID- 17114889 TI - Safety and efficacy of Juniperus ashei sublingual-swallow ultra-rush pollen immunotherapy in cypress rhinoconjunctivitis. A double-blind, placebo-controlled study. AB - BACKGROUND: The safety and efficacy of high-dose sublingual-swallow immunotherapy (SLIT) has been established in pollen rhinoconjunctivitis. This treatment has now been evaluated using an ultra-rush incremental dose regimen with a Juniperus ashei allergen extract in patients allergic to Cupressus sempervirens and Cupressus arizonica. METHODS: Patients received either placebo or SLIT. Evaluation of safety was based on the frequency of adverse events during the incremental dose period (half a day) and during maintenance therapy (4 months). Evaluation of efficacy was based on symptom and medication scores at the pollen peak. RESULTS: Seventy of the 76 patients included completed the study. There were no drop-outs during the rush procedure. One patient in the active group dropped out during the maintenance therapy due to adverse events: gastric pain and vomiting. There was also 1 drop-out in the placebo group due to pregnancy. Adverse events were infrequent, local and mild. Symptom scores for rhinitis and conjunctivitis were not statistically different between groups, but there was a marked and significant (p < 0.03) decrease of the medication score (about 50%) and nasal steroid consumption (about 75%) in the active treatment group. An increase from baseline of serum IgE and IgG4 J. ashei-specific antibodies was only observed in actively treated patients (p < 0.04 and p < 0.01, respectively). CONCLUSIONS: The tolerability and safety of high-dose ultra-rush SLIT were comparable to those reported in previous SLIT studies. SLIT with J. ashei extract, due to its high Jun a 1 content, significantly reduced nasal steroid consumption in patients allergic to European cypress. PMID- 17114890 TI - Allergic sensitization to common airborne allergens among adults in Estonia. AB - BACKGROUND: Allergies and asthma exhibit a growing health problem in the world. Sparse data are available about the prevalence of allergic sensitization in Estonia and Eastern Europe as well. We studied the prevalence of allergic sensitization, the associations between respiratory symptoms and allergic sensitization, and the influence of age, gender, area of residence, number of siblings and other demographic attributes on allergic sensitization in the population aged 17-69 years. METHODS: We conducted a cross-sectional study with 1,346 randomly selected participants distributed in 3 geographically and culturally distinct areas of Estonia. Sensitization to 15 aeroallergens was assessed by skin prick tests. The study also included a structured interview. RESULTS: The total prevalence of allergic sensitization was 33.0%. When estimated in patients aged 20-44 years, the prevalence was 38.5%. The most common sensitizer was cockroach followed by storage mites, while sensitization to cat, dog or pollen (common sensitizers in Scandinavian countries) was low. Allergic sensitization was significantly more common in urban and suburban compared with rural areas. Storage mites were the most common sensitizers in rural areas. Living in urban or suburban areas before the age of 5 significantly increased the risk for positive skin prick tests to several allergens. CONCLUSIONS: A higher prevalence of allergic sensitization than previously believed was found. Cockroach and storage mite allergens are suggested to be included in the routine investigation panel in Estonia. PMID- 17114891 TI - Differential inhibition of primary versus preactivated T cells by pimecrolimus but not by tacrolimus in vitro. AB - BACKGROUND: Recent studies in murine models of allergic contact dermatitis have shown that systemic treatment with pimecrolimus in contrast to tacrolimus did not inhibit the sensitization phase, whereas both compounds equivalently suppressed the inflammatory response in sensitized animals. This finding indicated a differential sensitivity of antigen-naive and primed T cells towards pimecrolimus and tacrolimus. METHODS: T cells obtained from healthy and allergic donors were subjected to primary and secondary stimulation by allogeneic or staphylococcal superantigen-presenting dendritic cells (DC). Human skin-derived, allergen specific T cell clones from an atopic dermatitis patient were activated by anti CD3 antibodies or by specific allergen-presenting DC. The inhibition of T cell proliferation and cytokine release by graded doses of calcineurin inhibitors was evaluated. RESULTS: Primary stimulation of T cells was inhibited by pimecrolimus with an approximately 8-fold lower potency as compared with tacrolimus. In contrast, the secondary response of ex vivo expanded T cells activated by allogeneic or staphylococcal superantigen-presenting DC was inhibited by both compounds with equivalent potency. Likewise, both drugs showed very similar potency to inhibit the proliferation and cytokine synthesis from antigen- stimulated T cell clones and the induction of cytokines in Jurkat T cells. CONCLUSION: These data indicate that pimecrolimus has a selectivity for antigen primed memory T cells not seen with tacrolimus. PMID- 17114892 TI - The bioinformatic study of transmembrane molecular transport. PMID- 17114893 TI - Phylogenetic analysis of general bacterial porins: a phylogenomic case study. AB - Bacterial porin proteins allow for the selective movement of hydrophilic solutes through the outer membrane of Gram-negative bacteria. The purpose of this study was to clarify the evolutionary relationships among the Type 1 general bacterial porins (GBPs), a porin protein subfamily that includes outer membrane proteins ompC and ompF among others. Specifically, we investigated the potential utility of phylogenetic analysis for refining poorly annotated or mis-annotated protein sequences in databases, and for characterizing new functionally distinct groups of porin proteins. Preliminary phylogenetic analysis of sequences obtained from GenBank indicated that many of these sequences were incompletely or even incorrectly annotated. Using a well-curated set of porins classified via comparative genomics, we applied recently developed bayesian phylogenetic methods for protein sequence analysis to determine the relationships among the Type 1 GBPs. Our analysis found that the major GBP classes (ompC, phoE, nmpC and ompN) formed strongly supported monophyletic groups, with the exception of ompF, which split into two distinct clades. The relationships of the GBP groups to one another had less statistical support, except for the relationships of ompC and ompN sequences, which were strongly supported as sister groups. A phylogenetic analysis comparing the relationships of the GenBank GBP sequences to the correctly annotated set of GBPs identified a large number of previously unclassified and mis-annotated GBPs. Given these promising results, we developed a tree-parsing algorithm for automated phylogenetic annotation and tested it with GenBank sequences. Our algorithm was able to automatically classify 30 unidentified and 15 mis-annotated GBPs out of 78 sequences. Altogether, our results support the potential for phylogenomics to increase the accuracy of sequence annotations. PMID- 17114894 TI - Engineering transport protein function: theoretical and technical considerations using the sugar-transporting phosphotransferase system of Escherichia coli as a model system. AB - Potential experimental approaches for developing and applying protein-engineering protocols to transmembrane transport systems are described. We specifically consider procedures designed to alter protein function. These procedures are designed for the specific purposes of (1) changing protein interaction specificities and (2) changing a protein's catalytic function. We use sugar transporting bacterial phosphotransferase systems as model systems to illustrate the proposed approaches. These and other similar procedures are likely to prove to be of utility for biotechnological manipulation of proteins as well as for elucidating potential evolutionary pathways taken for the appearance of novel functions within a protein family. PMID- 17114895 TI - MdfA from Escherichia coli, a model protein for studying secondary multidrug transport. AB - MdfA is a prototypic secondary multidrug transporter from Escherichia coli, which recognizes and exports a broad spectrum of structurally and electrically dissimilar toxic compounds. Here we review recent studies of MdfA, which, on the one hand, provide advanced understanding of certain aspects of secondary multidrug transport, and, on the other, address major mechanistic questions, some of which remain to be elucidated. Using biochemical, genetic, and physiological approaches, we have revealed several surprisingly promiscuous properties of MdfA including its multidrug recognition capacity, proton recognition determinants, aspects of energy utilization, and physiological role. PMID- 17114896 TI - Domain structure and pore loops in the 2-hydroxycarboxylate transporter family. AB - The 2-hydroxycarboxylate transporter (2HCT) family is a family of bacterial secondary transporters for substrates like citrate, malate and lactate. The family is in class ST[3] of the MemGen classification system that groups membrane proteins in structural classes based on hydropathy profile analysis. The combination of computational analysis of the proteins in class ST[3] and available experimental data on members of the 2HCT family has yielded a detailed structural model of the transporters. The core of the model is formed by two homologous domains with opposite orientation in the membrane. Each domain consists of 5 trans membrane segments and contains a pore loop between the 4th and 5th segment. The two pore loops enter the membrane-embedded part from opposite sides of the membrane (trans pore loops) and are believed to form the translocation pathway in the 3D structure. A genome wide study of the cellular location of the C-terminus of all Escherichia coli membrane proteins [Daley et al., 2005. Science 308:1321-1323] showed that the C-termini of the 19 E. coli proteins in class ST[3] were correctly predicted by the structural model. PMID- 17114897 TI - Calcium channel auxiliary subunits. AB - Many channels and carriers associate with auxiliary subunits which modify their activities and facilitate biogenesis. Advances in genome sequencing as well as biochemical, molecular genetic, and physiological experimentation have allowed for the discovery of many transport auxiliary subunits. Recent interests in the pharmacology of the calcium auxiliary subunits prompted a large amount of effort in deciphering their specific role in the conductance of calcium ions. In this review, we evaluate the functions of the 'extra' subunits of the voltage-gated calcium channels in animals as an example of auxiliary subunits of transporters in general. We discuss the functional data available for each of these subunits, present phylogenetic analyses, and discuss their potential evolutionary origins. Our analyses also reveal novel homologues of these subunits which might be of interest to the community. PMID- 17114898 TI - Topological predictions for integral membrane permeases of the phosphoenolpyruvate:sugar phosphotransferase system. AB - We report bioinformatic analyses of the largest superfamily of integral membrane permeases of the bacterial phosphotransferase system (PTS), the Enzyme IIC constituents of the Glc superfamily. Phylogenetic analyses reveal that this superfamily consists of five equally distant families, the Glucose (Glc), beta Glucoside (Bgl), Fructose (Fru), Mannitol (Mtl) and Lactose (Lac) families. Average hydropathy, amphipathicity and similarity plots were generated for these five families as well as for the entire superfamily. Charged residue distribution was analyzed, and the most conserved sequence motif, common to all five families, was identified. The results show that the members of all five families exhibit similar average hydropathy plots with regions of average amphipathicity and relative conservation also being similar. Evidence is presented suggesting that the Glucitol (Gut) family of Enzyme IIC constituents is a distant member of the Glc superfamily. Based on our analyses we offer a topological model that resembles, but differs in detail from the two previously proposed models. PMID- 17114899 TI - DNA fragmentation in chronic glomerulonephritis: an immunohistological analysis. AB - BACKGROUND: Experimental data suggest that apoptosis plays an important pathophysiological role in glomerulonephritis by restoring tissue structure after proliferation of intrinsic renal cells and infiltration of leukocytes. Relatively little is known of apoptosis in human glomerulonephritis, particularly in predicting renal function during follow-up. METHODS: In order to colocalize different markers for cell damage in renal tissue from patients with different forms of glomerulonephritis (GN), a series of semithin sections from 34 kidney biopsies were studied retrospectively. Normal kidney from a nephrectomy specimen with a small renal adenocarcinoma served as a control. DNA fragmentation, expression of tissue transglutaminase II, BAX and BCL-2 were visualized immunohistochemically. In some renal biopsies, immunohistochemical staining for activated caspase 3 was performed. Proinflammatory markers (C-reactive protein, leukocytes), serum creatinine, creatinine clearance, total proteinuria, albuminuria, alpha(1)-microglobulin and IgG excretion were determined at the time of biopsy. Serum creatinine and total proteinuria were assessed 6 and 12 months after renal biopsy. RESULTS: Nuclei with different degrees of DNA fragmentation were mainly found in epithelial cells of tubules, but also in glomerular cells, regardless of the form of GN studied. Transglutaminase II expression was found only in cells with a strong staining for DNA fragmentation. DNA fragmentation localized to glomerular cells was more pronounced in proliferative than in non proliferative forms of GN, being most abundant in patients with rapid progressive GN. Staining for activated caspase 3 in selected biopsies confirmed the presence of apoptosis. BAX and BCL-2 staining was detected within the same cells, but exhibited a different intracellular distribution. In proliferative GN, the extent of DNA damage in tubular epithelial cells significantly corresponds with the concentration of serum creatinine (p < 0.04) and with urinary excretion of alpha(1)-microglobulin (p < 0.01) at the time of biopsy. A significant correlation (p < 0.01) was seen between glomerular DNA fragmentation and follow up total proteinuria 12 months after biopsy for proliferative forms of GN. The damaged glomerular area (e.g. mesangial sclerosis) significantly correlated with DNA fragmentation in proliferative, but not in nonproliferative GN at the time of biopsy. Furthermore, glomerular damaged showed a significant correlation with tubular DNA damage in proliferative GN. CONCLUSION: In glomerular cells, apoptosis may be important for the clearance of proliferating cells whereas in tubules, cell damage showed dependence on the degree of tubular injury mediated by inflammation and/or proteinuria. Although the degree of apoptosis in tubular cells correlates with serum creatinine in proliferative GN at the time of biopsy, it is of limited use to predict future renal function. PMID- 17114900 TI - Hemoglobin changes at the initiation of high-flux hemodialysis. AB - BACKGROUND: The aim of the present study was to assess hemoglobin changes occurring at the beginning of high-flux hemodialysis (HD). METHODS: In a group of 20 chronic HD patients (group A), total hemoglobin (tHb), hematocrit (Hct) and total serum proteins (TP) were measured in blood samples drawn from an arterial fistula needle before the initiation of high-flux HD, and from an arterial line 5 min after HD with the dialysate in the bypass mode. 31 chronic stable HD patients (group B) served as controls. In group B patients, tHb was measured in blood samples drawn from an arterial fistula needle before the initiation of high-flux HD, and from arterial and venous lines simultaneously 5 min later. Blood flow rates in groups A and B were set from the beginning of the study to 300 ml/min, while the bicarbonate dialysate flow rate and ultrafiltration rate in group B patients was set to 700 ml/min and zero, respectively. The same high-flux dialyzer was used for all patients (FLX-18, membrane PEPA 1.8 m(2)). RESULTS: A comparison of baseline (pre-dialysis) values with those derived from an analysis of the arterial line in groups A and B at 5 min revealed that tHb decreased by 0.6 +/- 0.2 g/dl (5.2 +/- 1.7%, p < 0.001) and 0.7 +/- 0.7 g/dl (5.4 +/- 6.2%, p < 0.001), respectively. At the same time, Hct and TP in group A decreased by 1.32 +/- 0.7% (3.8 +/- 2.0%, p <0.001) and 0.3 +/- 0.1 g/dl (4.8 +/- 1.4%, p < 0.001), respectively. Blood volume (BV) and plasma volume (PV) in group A patients at 5 min as calculated from tHb and TP values increased by 5.6 +/- 1.9 and 5.2 +/- 1.7%, respectively, while BV in group B patients increased by 6.1 +/- 7.0% (not significant when compared to group A). tHb did not change significantly in 14 patients (group C) studied immediately after adopting the supine position and 5 min later in the absence of HD. CONCLUSION: A 5% decrease in tHb was observed 5 min after the initiation of high-flux HD with a zero ultrafiltration rate, and was due to an increase in BV. PMID- 17114901 TI - Sleep disorders are underdiagnosed in patients on maintenance hemodialysis. AB - BACKGROUND: Sleep apnea-hypopnea syndrome (SAHS) is a cardiovascular risk factor. The aim of this study was to evaluate sleep disorders using polysomnography on a non-selected population of patients on maintenance hemodialysis. METHODS: Overnight polysomnography was performed on 32 hemodialysis patients (24 men/8 women, 54 +/- 16 years), and on 19 healthy subjects of similar age, sex and body mass index who were used as controls. RESULTS: In hemodialysis patients, the most frequent sleep disorder was SAHS in 44% (14/32), followed by insomnia in 41% (13/32). Compared to healthy controls, patients on hemodialysis showed less slow wave sleep and rapid eye movement sleep (23 vs. 36%, p = 0.001), less sleep efficiency (71 vs. 87%, p = 0.0079) and a higher periodic limb movement index (39.7 vs. 9.1; p = 0.003). An increase in apnea-hypopnea index (18.9 vs. 4.3; p = 0.007) and dips in the SaO(2) (> or =4%) per hour of sleep (22.6 vs. 6.4; p = 0.021) were also significantly greater in hemodialysis patients than controls. 72% of the cases of SAHS were diagnosed solely by means of polysomnography. CONCLUSIONS: The patients on hemodialysis showed poor sleep quality with a significant increase in the apnea-hypopnea index and in the number of dips in SaO(2). SAHS was underdiagnosed in a large percentage of the hemodialysis patients. PMID- 17114902 TI - Total nitrogen and free amino acid losses and protein calorie malnutrition of hemodialysis patients: do they really matter? AB - BACKGROUND/AIMS: Protein calorie malnutrition (PCM) in patients on hemodialysis (HD) is multifactorial; however, HD per se induces nutrient losses. The aim of the present study was to characterize the losses of total nitrogen (TN) and free amino acids (FAs) through the dialysate and to determine the relationship between this loss and PCM, food ingestion, and the characteristics of the hemodialyzer in patients on HD. METHODS: In a prospective study, 21 patients submitted to low flux HD 3 times a week were evaluated within a period of 6 months regarding nutritional status, dietary calorie and protein intake, and losses through the dialysate of TN, FA, and urea nitrogen (UN). The type, surface area and reuses (up to 12) of the dialysis membrane were determined on each occasion, and the adequacy of dialysis was estimated by Kt/V. RESULTS: 50% of the patients were considered malnourished, although the mean protein and energy intakes were similar for the malnourished and nourished patients. Mean TN losses through the dialysate were 16 g/session (60% UN). FA losses varied from 3.8 to 4.2 g/total volume. TN and FA in the dialysate did not differ significantly between malnourished and nourished patients. There was a positive correlation between membrane (polysulfone) area and TN (p <0.05) and ultrafiltrate volume and TN (p < 0.05), and a nonsignificant correlation between reuse of the dialysis membrane and TN. CONCLUSIONS: TN and FA losses through the dialysate were similar for malnourished and non-malnourished patients on chronic HD, thus they do not act as indicators of nutritional status impairment. PMID- 17114903 TI - Anandamide inhibits endothelin-1 production by human cultured endothelial cells: a new vascular action of this endocannabinoid. AB - The endogenous cannabinoid receptor agonist anandamide (AEA) exerts vascular effects such as vasodilatation and hypotension. In this study, we determined the effect of AEA on endothelin-1 production by cultured human umbilical vein endothelial cells. Anandamide (>or=5 micromol/l) significantly decreased endothelin-1 production in a dose-dependent manner, a response not affected by the specific CB1 receptor antagonist/inverse agonist SR-141716A. Adenosine, via activation of adenosine receptors (also targets for SR-141716A), was not involved in these effects. Conversely, AEA increased nitric oxide (NO) production, an effect inhibited by SR-141716A, indicating the involvement of CB1 receptors. Therefore, we hypothesize that AEA effects on endothelial cells may lead to vasodilatation through independent concerted mechanisms, involving a non-CB1 receptor-dependent inhibition of endothelin-1 production and a CB1-mediated increase of NO. PMID- 17114904 TI - Unacylated as well as acylated ghrelin promotes cell survival and inhibit apoptosis in HIT-T15 pancreatic beta-cells. AB - Ghrelin is mainly produced by the stomach, although it is expressed in other tissues, including the pancreas. Among its pleiotropic actions, ghrelin prevents the development of diabetes in rats and exerts mitogenic and antiapoptotic effects in different cell types. In addition, a ghrelin-producing epsilon-cell population has been demonstrated in rodent islets, suggesting a direct role in the control of islet cell survival. In this study, we investigated the effect of acylated ghrelin (AG) and unacylated ghrelin (UAG) on cell survival of HIT-T15 pancreatic beta cells. We show that both AG and UAG equally prevented beta cell death induced by serum withdrawal. In addition, both peptides inhibited serum starvation-induced apoptosis. These findings indicate that UAG and AG prevent cell death and apoptosis of pancreatic beta cells. Since only AG, but not UAG, binds the GRLN receptor, a different and as yet unknown receptor is likely involved in these survival mechanisms. PMID- 17114905 TI - A randomized controlled trial to evaluate the efficacy of ultrasound-guided laser photocoagulation for treatment of benign thyroid nodules. AB - This randomized controlled study was designed to test the efficacy and safety of percutaneous ultrasound (US)-guided laser photocoagulation (PLP) for treatment of subjects with compressive symptoms due to benign thyroid nodules and/or at high surgical risk. Twenty six subjects were randomized to the intervention (no. 13, age 68+/-3 yr, mean+/-SEM) or observation (no. 13, age 71+/-2 yr) groups. In the control group, the volume of nodules did not significantly change over the 30 week period of observation. In the intervention group, median nodule volume at baseline was 8.2 ml (range 2.8-26.9) and was not significantly different from that of the control group. Nodules decreased significantly (p<0.0001) by 22% after 2 weeks (6.5 ml; range 2.4-16.7) and by 44% after 30 weeks (4.6 ml; range 0.69-14.2). Energy given was correlated (p<0.05) with the reduction of thyroid nodule volume. All patients tolerated the treatment well and reported relief from compressive and cosmetic complaints (p<0.05). At the time of enrolment 7/13 (54%) and 6/13 (46%) of patients in the intervention and control groups, respectively, had sub clinical hyperthyroidism. PLP normalized thyroid function at 6 and 30 weeks after treatment. In conclusion, PLP is a promising safe and effective procedure for treatment of benign thyroid nodules in patients at high surgical risk. PMID- 17114906 TI - Long-term effects of elevated gastrin levels on calcitonin secretion. AB - Gastrin and pentagastrin stimulate thyroid C cells and elevate serum calcitonin levels. Gastrin levels may be elevated when histamine-2 receptor blockers (H2RB) and/or proton pump inhibitors (PPI) are used, or in patients with pernicious anemia (PA). This study was designed to investigate the long-term effects of elevated gastrin levels on serum calcitonin levels. We conducted a pentagastrin stimulation test to evaluate C cell reserves in patients who had been using PPI and/or H2RB for an extended period, as well as in patients with PA. We compared the results with a healthy control group of similar age and sex. A total of 40 controls (26 women, 24 men) and 25 patients (15 women, 10 men) using H2RB and/or PPI, and 37 patients (24 women, 13 men) with PA were enrolled. The groups were similar in terms of mean age and sex distribution. Mean fasting gastrin levels, and mean baseline and pentagastrin-stimulated calcitonin levels were significantly higher in the H2RB+PPI and PA groups than in controls. No significant differences were observed between the H2RB+PPI and PA groups. There was no correlation between gastrin and calcitonin levels. However, mean calcitonin levels were significantly higher in subjects with high baseline gastrin levels than in controls. The prevalence of autoimmune thyroid disease was 32% in the PA group. Patients with PA and thyroiditis had significantly higher baseline gastrin levels than patients with PA only (p<0.01). PA with autoimmune thyroid disease had also significantly higher baseline and pentagastrin stimulated calcitonin levels than did PA patients without autoimmune thyroid disease. In conclusion, chronic elevated gastrin levels led to elevated calcitonin levels. Further histopathological studies showing C cell hyperplasia are needed to confirm the mechanism of this relationship. PMID- 17114907 TI - Lack of association between the tetranucleotide repeat polymorphism in the 3' flanking region of the leptin gene and hypertension in severely obese patients. AB - Conflicting data suggest an association between leptin gene polymorphisms and essential hypertension independently of obesity. The aim of this study was to evaluate, in severely obese subjects, the role of one of these polymorphic markers in relation to the development of hypertension. The study included 325 obese patients with mean body mass index (BMI) of 46+/-6.94 kg/m2. One hundred sixty-six were hypertensive and 159 normotensive. In both groups, the presence of a tetranucleotide repeat in the 3' flanking region of the Ob gene was investigated using polymerase chain reaction (PCR). Due to the genetic variant, in the region studied it is possible to distinguish two alleles with different size distribution: Class I (shorter one) and Class II (longer one). Class I and Class II allele frequencies were not significantly different in obese patients when analyzed according to the presence or absence of hypertension. The results presented herein do not support a significant association of this Ob gene polymorphism with hypertension. These findings are in contrast with that reported in other populations. However, we cannot rule out that different ethnicity and/or phenotypic variability might mask small effects. PMID- 17114908 TI - Ghrelin and cortistatin in lung cancer: expression of peptides and related receptors in human primary tumors and in vitro effect on the H345 small cell carcinoma cell line. AB - Ghrelin, a natural GH secretagogue (GHS) acylated peptide, and cortistatin (CST), a natural SRIF-like peptide, interfere with neoplastic growth in different cancers. We tested forty-one lung carcinomas and the H345 small cell lung carcinoma (SCLC) cell line by RT-PCR to investigate the presence of ghrelin and CST and related receptors, including type 1a GHS receptor (GHS-R1a), all SRIF receptor subtypes (sst 1-5) and MRGX2. Moreover, the presence of ghrelin and CST peptides was studied in both tumors and H345 cells. Ghrelin and CST mRNA were present in the majority of tested tumors, but ghrelin and CST proteins were revealed only in tumors with a neuroendocrine phenotype. All the receptors mRNA had a heterogeneous expression without correlation between ghrelin (or CST) and their receptor distribution. All the transcripts, but not GHS-R1a, were expressed in H345 cells. However, ghrelin and desacyl ghrelin induced in vitro a dose dependent inhibition on the H345 cell proliferation and increased apoptosis. Conversely, neither CST nor SRIF affected H345 cell growth, despite the presence of their specific receptors. The anti-proliferative and the pro-apoptotic effects of ghrelin were consistent with binding experiments on H345 cell, where either acylated or des-acylated ghrelin recognized a common binding site. In conclusion, the present study indicates that: a) ghrelin and CST mRNAs are expressed in lung cancers, although some neuroendocrine tumors contain detectable amounts of the peptides; b) GHSR-1a mRNA is present exclusively in neuroendocrine tumors, whereas MRGX2 mRNA (but not peptide) is expressed in all histological types; c) both ghrelin forms inhibit H345 cell proliferation, both directly and enhancing apoptosis, despite the absence of GHS-R1a, whereas CST and its receptors do not interfere with cell growth. PMID- 17114909 TI - Oxidative stress in the metabolic syndrome. AB - The metabolic syndrome represents a cluster of several risk factors for atherosclerosis that increases the risk of future cardiovascular events. In this study, we evaluated whether oxidative stress is increased in subjects with the metabolic syndrome. We studied 100 subjects (50 men and 50 women) with the metabolic syndrome, as defined by the Adult Treatment Panel III, and 50 (25 men and 25 women) matched subjects without the syndrome. Insulin sensitivity was assessed with the homeostasis model assessment (HOMA) methods; endothelium dependent flow-mediated vasodilation (FMD) was evaluated in the right brachial artery with a high-resolution ultrasound machine; oxidative stress was assessed by measuring the circulating levels of nitrotyrosine (NT), considered a good marker for the formation of endogenous peroxynitrite. Compared with control subjects, patients with the metabolic syndrome had greater waist circumference, higher HOMA and systolic pressure values, higher triglyceride and lower HDL cholesterol levels. NT levels were higher (0.44+/-0.12 micromol/l, mean+/-SD) while FMD was lower [7.3 (4.4/9.6), median and interquartile range] in subjects with the metabolic syndrome as compared with control subjects [0.27+/-0.08 and 11.8 (8.6/14.9), respectively, p<0.001]. There was an increase in NT levels and HOMA score as the number of components of the metabolic syndrome increased. NT levels were associated with waist circumference (r=0.38, p=0.01), triglycerides (r=0.32, p<0.02), systolic blood pressure (r=0.21, p<0.05) and fasting glucose (r=0.24, p<0.05). The oxidative stress that accompanies the metabolic syndrome is associated with both insulin resistance and endothelial dysfunction, providing a connection which is highly deleterious for vascular functions. PMID- 17114910 TI - Cortisol, dehydroepiandrosterone sulphate and dehydroepiandrosterone sulphate/cortisol ratio responses to physical stress in males are influenced by pubertal development. AB - To evaluate the influence of chronological age and pubertal development on the hypothalamus-pituitary-adrenal (HPA) axis response to stress, we studied the possible correlations between male pubertal characteristics and salivary cortisol (C), DHEAS and the DHEAS/C ratio before (pre-stress) and after acute exercise stress in young male volunteers (no. 87; 13.3+/-2.1 yr). In our overall study population, the mean pre-stress salivary C and DHEAS concentrations, significantly increased after exercise-related stress, whereas the DHEAS/C ratio significantly decreased. Pre-stress salivary C was positively correlated with chronological age, and after-stress salivary C concentration variations were negatively correlated with pubertal stage, mean testis volume and pre-stress salivary DHEAS. Furthermore, salivary DHEAS concentrations and the DHEAS/C ratio, before and after exercise stress, were positively correlated with chronological age, pubertal stage, pre-stress salivary testosterone (T), testis volume and body mass index (BMI). In contrast with late pubertal stages (P4, P5), young individuals at early stages of puberty (P1 to P3) showed higher C increase and lower DHEAS/C ratio after exercise-related stress. In conclusion, since C is also a mediator of stress-related negative effects on health and the DHEAS/C ratio has been hypothesized as an index for the degree to which an individual is buffered against the negative effects of stress, these data might suggest potentially increased stress-related risks at early stages of male puberty. PMID- 17114911 TI - GH response to hypoglycemia and clonidine in the GH-releasing hormone resistance syndrome. AB - GH secretion by the pituitary is the result of the balance between the stimulatory effect of GHRH and the inhibitory effect of SS. Patients with mutations in GHRH receptor (GHRH-R) gene (GHRH-R) offer a unique model to study the mechanism of action of different GH secretion stimuli. In the past, we have demonstrated a small but significant GH response to a GH secretagogue (GHRP-2) in a homogenous cohort of patients with severe GH deficiency (GHD) due to a homozygous null mutation in GHRH-R (IVS1+1G-->A). Now, we sought to determine if we could detect a GH response to hypoglycemia (ITT: insulin tolerance test) or clonidine (CL) in these patients. Nine young GHD subjects underwent both ITT and CL tests, and 2 additional subjects underwent only CL test. There was a small but significant GH increase during ITT, but not during CL test. These results indicate that a minimal albeit significant GH response to ITT can occur despite complete lack of GHRH-R function. PMID- 17114912 TI - High prevalence of secondary hyperparathyroidism due to hypovitaminosis D in hospitalized elderly with and without hip fracture. AB - OBJECTIVES: To determine and compare the prevalence of secondary hyperparathyroidism (HPTH) in a population of community-dwelling and institutionalized older adults hospitalized with and without hip fracture, and to evaluate factors correlated with secondary HPTH in this population. METHODS: Circulating concentrations of serum intact PTH, 25-hydroxyvitamin D [25(OH)D] total serum calcium and albumin were measured in 160 subjects with an osteoporotic fracture of the proximal femur and in 160 matched controls hospitalized for a disease unrelated to bone status. Patients with secondary causes of bone loss and taking medications affecting bone metabolism were excluded. Age, sex, place of residence and the ability to perform basic activities of daily living (BADL) two weeks before hospital admission were recorded at baseline. RESULTS: Patients were comparable with regard to the baseline demographic, biochemical and functional characteristics. The overall prevalence of secondary hyperparathyroidism was 51.2%, without significant differences between hip fractured patients and controls (50.6 vs 51.9%, p=0.911). In bivariate analysis only the age and functional status (BADL) demonstrated a significant relationship with secondary HPTH, while sex and place of residence were not significant. These results were also confirmed in multivariate analysis. Particularly, the risk of secondary HPTH increased with age and with the number of functions lost in BADL: patients fully dependent showed a 3 times as high risk (odd ratio 3.07, 95% confidence interval 1.73 to 5.46, p=0.000) compared to patients independent in BADL, and subject aged >88 yr had a twice as high risk of developing secondary HPTH compared to younger ones (odd ratio 2.28, 95% confidence interval 1.20 to 4.32, p=0.012). CONCLUSION: These results show that secondary HPTH due to hypovitaminosis D is a frequent disorder in hospitalized elderly, strongly correlated with the functional status, irrespective of sex and place of residence. PMID- 17114913 TI - Methylenetetrahydrofolate reductase gene polymorphism, homocysteine and risk of macroangiopathy in Type 2 diabetes mellitus. AB - OBJECTIVE: A polymorphism in the gene for methylenetetrahydrofolate reductase (MTHFR) has been reported to be associated with hyperhomocysteinemia and risk for atherosclerotic vascular diseases. In this case-control study, we examined the distribution of the MTHFR genotypes in the Chinese population and clarified the relationship between the gene polymorphism for MTHFR and macroangiopathy in Chinese Type 2 diabetes mellitus. METHODS: Two hundred and sixteen unrelated patients with Type 2 diabetes mellitus, 112 of whom had macroangiopathy, and 114 healthy control subjects, were recruited. The MTHFR C677T genotype was analyzed by polymerase chain reaction-restriction fragment length polymorphism. Plasma total homocysteine levels were measured using high-performance liquid chromatography (HPLC) with fluorescence detection. RESULTS: In 114 healthy control subjects, the frequency of the mutant T allele was 31.1%. The genotype distribution did not differ between control subjects and Type 2 diabetic patients (chi2=3.03, p=0.220). Genotypic analysis revealed that the MTHFR genotype was different between diabetic patients with and without macroangiopathy (chi2=12.42, p=0.002). Type 2 diabetic patients with macroangiopathy displayed a greater prevalence of T allele than Type 2 diabetic patients without macroangiopathy (44.6 vs 29.3%; chi2=10.82, p=0.001). The odds ratio for macroangiopathy in Type 2 diabetic patients in presence of T allele was 1.94 [confidence interval (CI) 95%: 1.31-2.89]. Moreover, plasma homocysteine levels were markedly higher in individuals with TT genotype than those with CC or CT genotype. CONCLUSIONS: The C677T mutation of MTHFR gene is common in the Chinese population. MTHFR C677T gene polymorphism associated with a predisposition to increased plasma homocysteine levels could constitute a useful predictive marker for macroangiopathy in Chinese Type 2 diabetic patients. PMID- 17114914 TI - Development of acromegaly in a patient with anorexia nervosa: pathogenetic and diagnostic implications. AB - Anorexia nervosa (AN) is a psychosomatic disorder characterized by an alteration in body image, resulting in eating abnormalities that lead to malnutrition associated with secondary endocrinological disturbances. AN is associated with high levels of GH and low levels of IGF-I, suggestive of a nutritionally acquired lack of GH action or GH resistance and of an unbalanced hypothalamic GH neuroregulation with GHRH prevalence on SS tone. In this clinical report, we describe the development of acromegaly in a patient affected by AN and we discuss pathogenetic and diagnostic implications. PMID- 17114915 TI - Dissociation between tumor shrinkage and hormonal response during somatostatin analog treatment in an acromegalic patient: preferential expression of somatostatin receptor subtype 3. AB - INTRODUCTION: About a third of acromegalic patients is resistant to available SS analogs (SA), octreotide (OCT) and lanreotide (LAN). Such resistance is related to reduction of SS receptor (SSTR) density or to a different expression of SSTR subtypes. There are 5 known SSTR subtypes. SSTR2 and SSTR5 are usually expressed in GH-secreting pituitary tumors, and both SA bind preferentially to SSTR2 and, to a lesser extent, to SSTR5. We herein describe an acromegalic patient who presented impressive tumor shrinkage without hormonal normalization during primary therapy with SA. MATERIAL AND METHODS: This 23-yr-old male acromegalic patient was treated with slow-release LAN (LAN-SR), 30 mg every 10 days for six months, followed by OCT-LAR, 30 mg every 28 days for an additional six months with a 75% tumor volume reduction but without GH and IGF-I normalization. Subsequently, he underwent pituitary surgery and expression of SSTR in the removed tumor was performed by real time RT-PCR by the 2-deltaCt method, using GAPDH as internal control. All PCR products were confirmed by automated sequencing. RESULTS: SSTR expression revealed an unusual profile, with almost exclusively expression of SSTR3. CONCLUSIONS: These unusual clinical and receptor subtypes profile suggest an important role of SSTR3 on tumor shrinkage. The low affinity of LAN and OCT for this SSTR subtype could be compensated by its high expression in this GH-secreting pituitary macroadenoma. PMID- 17114916 TI - Occurrence of overt celiac disease in the elderly following total thyroidectomy. AB - We report the case of a female patient in whom gluten-induced entheropathy was revealed at the age of 71 yr by resistance to treatment with levothyroxine (L T4), calcium carbonate and alfacalcidol. Hypothyroidism and hypoparathyroidism were the consequence of a total thyroidectomy performed at the age of 65 yr for a large multinodular goiter. Six months after thyroid ablation the patient started to complain of abdominal pain, diarrhea and weight loss. Following, anemia and osteopenia were documented. A progressive increase of replacement therapy for hypothyroidism and hypoparathyroidism was necessary. The clinical presentation suggested a malabsorption syndrome: celiac disease (CD) was diagnosed by serological markers and duodenal biopsy. Following gluten-free diet a normalization of clinical and serological findings was observed, bone mass density improved and a reduction of L-T4, calcium and vitamin D requirements was observed. PMID- 17114917 TI - Sarcoidosis of the thyroid gland associated with hyperthyroidism: review of the literature and report of two peculiar cases. AB - Sarcoidosis is a systemic disease characterized by non-caseating granulomas that rarely involve the thyroid gland. Thyroid sarcoidosis has seldom been documented, and few cases have so far been described in association with hyperthyroidism. Here, we review the literature on this association, report two patients presenting with hyperthyroidism and histologically-proven sarcoidosis, and discuss related clinical, biochemical, pathological and genetic findings. PMID- 17114918 TI - How to localize parathyroid tumors in primary hyperparathyroidism? AB - The management of primary hyperparathyroidism (PHPT) has dramatically changed in the last 5 yr. Many more patients now undergo focused, limited or minimally invasive parathyroidectomy instead of traditional bilateral neck exploration. This change has taken place because of the improved accuracy of pre-operative localizing studies in selecting patients who have single-gland parathyroid disease (single adenoma) and can therefore have a minimally invasive parathyroidectomy. Sestamibi scanning followed by ultrasound, magnetic resonance imaging (MRI) and computed tomography (CT) scans are most accurate for localizing parathyroid tumors in patients with PHPT. Selective venous catheterization for PTH levels is useful when other localizing studies are negative or discordant in patients with persistent or recurrent PHPT. The routine use of one or more localizing studies commonly identifies the parathyroid tumor in patients with single-gland disease; but if localizing studies are negative or discordant, patients should have intra-operative PTH levels monitored or have a bilateral neck exploration to ensure a high rate of biochemical cure. PMID- 17114919 TI - Reduced need of glucocorticoid therapy in a woman with congenital adrenal hyperplasia due to 21-hydroxylase deficiency during pregnancy. PMID- 17114920 TI - A novel heterozygous deletion frameshift mutation of GATA3 in a Japanese kindred with the hypoparathyroidism, deafness and renal dysplasia syndrome. PMID- 17114921 TI - How should the "optimal" pre-operative localizing imaging work-up in hyperparathyroid patients be? PMID- 17114922 TI - Regulation of Unr expression by 5'- and 3'-untranslated regions of its mRNA through modulation of stability and IRES mediated translation. AB - Unr (upstream of N-ras) is a cytoplasmic RNA-binding protein that can act as a regulator of mRNA stability and IRES-mediated translation. Unr, a member of the cold-shock domain (CSD) protein super-family, is ubiquitously expressed, with variable abundance, in different tissues or during embryonic development. Prokaryotic and eukaryotic cold-shock protein expression is highly regulated at both the transcriptional and post-transcriptional levels. Here we analyzed the role of the 5'- and 3'-untranslated regions (UTR) of unr mRNA in post transcriptional regulation of Unr expression. We show that, in vitro, unr 3'-UTR specifically destabilizes unr transcripts. Accordingly, in vivo, the half-life of unr messages deleted of noncoding regions is increased by approximately 3.6 fold, resulting in an enhanced steady-state level of Unr protein. We also show that the 5'-UTR exhibits IRES activity both when translated in vitro and in transiently transfected cells. This IRES activity displays cell type specificity with a higher efficiency in HeLa and HuH7 than in ES cells. Moreover, Unr IRES activity was higher in unr(-/-) than in unr(+/+) ES cells, indicating that Unr negatively regulates its own IRES activity. Our studies further reveal that Unr specifically interacts with its own mRNAs in vivo. These results suggest that a feedback control of mRNA translation is involved in regulating Unr expression. PMID- 17114923 TI - Rational probe optimization and enhanced detection strategy for microRNAs using microarrays. AB - MicroRNAs (miRNAs) are post-transcriptional regulators participating in biological processes ranging from differentiation to carcinogenesis. We developed a rational probe design algorithm and a sensitive labelling scheme for optimizing miRNA microarrays. Our microarray contains probes for all validated miRNAs from five species, with the potential for drawing on species conservation to identify novel miRNAs with homologous probes. These methods are useful for high-throughput analysis of micro RNAs from various sources, and allow analysis with limiting quantities of RNA. The system design can also be extended for use on Luminex beads or on 96-well plates in an ELISA-style assay. We optimized hybridization temperatures using sequence variations on 20 of the probes and determined that all probes distinguish wild-type from 2 nt mutations, and most probes distinguish a 1 nt mutation, producing good selectivity between closely-related small RNA sequences. Results of tissue comparisons on our microarrays reveal patterns of hybridization that agree with results from Northern blots and other methods. PMID- 17114924 TI - Specific RNAi mediated gene knockdown in zebrafish cell lines. AB - Here we demonstrate highly efficient RNA interference in ZFL, SJD and ZF4 cell lines derived from adult and embryonic zebrafish Danio rerio. Microinjection of siRNAs resulted in silencing in almost 100% of cells while transfection using cationic liposomes led to silencing in 30%. Use of siRNAs against zebrafish lamin A, lamin B2 and the motor protein Eg5, led to knockdown of the target genes with the specific phenotypes expected from prior studies in mammalian cells. In contrast injection of lamin A, GL2 and eGFP siRNAs into zebrafish embryos resulted in morphological defects, abnormal development and early death of most embryos. The results indicate unspecific responses to siRNAs in the embryo but a fully developed and active RNAi machinery in cell lines. PMID- 17114925 TI - A cullin E3 ubiquitin ligase complex associates with Rik1 and the Clr4 histone H3 K9 methyltransferase and is required for RNAi-mediated heterochromatin formation. AB - The assembly of heterochromatin in fission yeast and metazoans requires histone H3-lysine 9 (-K9) methylation by the conserved Clr4/Suv39h methyltransferase. In fission yeast, H3-K9 methylation requires components of the RNAi machinery and is initiated by the RNA-Induced Transcriptional Silencing (RITS) complex. Here we report the purification of a novel complex that associates with the Clr4 methyltransferase, termed the CLRC (CLr4-Rik1-Cul4) complex. By affinity purification of the Clr4-associated protein Rik1, we show that, in addition to Clr4, Rik1 is associated with the fission yeast E3 ubiquitin ligase Cullin4 (Cul4, encoded by cul4(+)), the ubiquitin-like protein, Ned8, and two previously uncharacterized proteins, designated Cmc1 and Cmc2. In addition, the complex contains substochiometric amounts of histones H2B and H4, and the 14-3-3 protein, Rad24. Deletion of cul4(+), cmc1(+), cmc2(+) and rad24(+) results in a complete loss of silencing of a ura4(+) reporter gene inserted within centromeric DNA repeats or the silent mating type locus. Each of the above deletions also results in accumulation of noncoding RNAs transcribed from centromeric repeats and telomeric DNA regions, and a corresponding loss of small RNAs that are homologous to centromeric repeats, suggesting a defect in the processing of noncoding RNA to small RNA. Based on these results, we propose that the components of the Clr4 Rik1-Cul4 complex act concertedly at an early step in heterochromatin formation. PMID- 17114926 TI - A loxP-containing pol II promoter for RNA interference is reversibly regulated by Cre recombinase. AB - Several DNA vectors for RNA interference in mammalian cells have been described. These express a short hairpin RNA (shRNA) that is subsequently processed into mature small interfering RNAs (siRNAs). We previously developed the siRNA expressing vector psiUx based on the polII promoter of the U1 small nuclear RNA gene. Here we describe the conversion of such construct into an inducible system. The starting construct psiUStuff contains a loxP-Stuffer-loxP cassette just upstream the transcription initiation site and does not express the shRNA until the two canonical loxP sites undergo Cre-mediated recombination. If sustained expression of the recombinase is maintained, transcription is repressed and shRNA synthesis is abolished. Therefore, in our system the Cre recombinase exhibits the dual function of activator and repressor allowing the on/off regulation of siRNAs production. Using a Cre recombinase whose transcription is under the control of a tetOn system, we show the temporally controlled expression of an shRNA directed towards the lamin A/C mRNA, as well as the regulated knockdown of its target. PMID- 17114927 TI - Determination and analysis of the pre-mRNA cleavage sites in Arabidopsis. AB - Alignment of the Arabidopsis cDNA to genome DNA sequences revealed that approximately half of the mRNAs were found to contain 3' non-templated nucleotide addition prior to the poly(A) sequence. These findings suggest that we can more precisely determine the cleavage sites. Based on the nucleotide downstream of cleavage sites, we are able to derive a hierarchy of cleavage preferences A>>U>C>>G. Interestingly, the completely different hierarchy of preferences was derived to be U>>A>>G>C, based on the nucleotide upstream of cleavage sites. Dinucleotide AU is the most preferred composition for cleavage. The determination of the cleavage sites and systematical analysis can help in the enhancement of 3' processing site prediction and mechanistic understanding of 3'-end processing in plant. PMID- 17114928 TI - Optimal random libraries for the isolation of catalytic RNA. AB - The relationship between ribozyme size and catalytic activity is of fundamental importance for RNA catalysis and molecular evolution in the RNA world. We have performed a series of competitive in vitro selection experiments to probe the relationship using RNA libraries containing size-heterogeneous random regions. Our experiments have established an inverse correlation between RNA replication efficiency (the combined efficiency of PCR amplification, transcription, and reverse transcription) and RNA size. A number of ribozyme sequences have been isolated from different RNA size groups under competitive selection conditions. Comprehensive kinetic analysis on isolated ribozymes has revealed that large ribozymes do not confer a significant catalytic superiority over smaller ones under most selection conditions, and actually impose two significant problems of replication inefficiency and RNA misfolding into inactive conformations. The fraction of a misfolded ribozyme population is defined as alpha. Large ribozymes tend to possess high alpha values, which may significantly reduce ribozyme performance. Our results suggest that a random region of around 60 nucleotides represents the optimal balance between ribozyme catalytic activity, RNA misfolding (alpha), and replication efficiency, and may therefore constitute the most advantageous RNA libraries for successful isolation of functional RNA sequences. PMID- 17114929 TI - Redesign of an artificial ligase ribozyme based on the analysis of its structural elements. AB - The catalytic and folding properties of "DSL ribozyme" were investigated. This artificial ligase ribozyme was constructed by installing a catalytic unit to a designed self-folding RNA. The self-folding RNA was composed of three helices connected via two tertiary interactions that served as scaffolding in the molecular design. The present analysis revealed that the tertiary interaction between the GAAA loop and its specific receptor plays a crucial role in the folding of the active structure and the precise positioning of the catalytic site. On the basis of the analyses, the ribozyme was redesigned and converted to two advanced forms--a smaller derivative with appreciable catalytic activity and a derivative with RNA polymerase-like activity. The study demonstrates that redesign of an artificial ribozyme is effective and efficient if its structural elements are finely resolved. This kind of molecular transformation should serve as a prototypic model for understanding the molecular organization and evolution of naturally occurring ribozymes. PMID- 17114930 TI - Variable splicing of non-coding roX2 RNAs influences targeting of MSL dosage compensation complexes in Drosophila. AB - The non-coding roX1 and roX2 RNAs are components of the MSL dosage compensation complex in Drosophila. We found that multiple species of roX2 RNA are produced by alternative splicing, with one major and at least 20 different minor forms associated with MSL proteins. The alternative forms are generated by variable usage of multiple 5' and 3' splice sites between two common exons. This alternative splicing is evolutionarily conserved in several distant Drosophila species in spite of differences in primary sequences. Transgenic constructs expressing individual major or minor D. melanogaster roX2 species display low steady-state levels of roX2 RNA, weak accumulation of MSL complex on the X chromosome, and low rescue of male-specific roX(-) lethality. Increased expression of individual roX2 forms using the constitutive Hsp83 promoter results in increased transgenic rescue of roX(-) mutant male flies. However, although males survive they are delayed in their development. In addition, MSL complexes still show low affinity for the X chromosome and abnormal accumulation at the transgenic site of synthesis of the individual roX2 alternative splice form. Taken together, these results suggest an important role for roX2 RNA splicing in optimal MSL complex assembly or function. PMID- 17114931 TI - Hypomodification of transfer RNA in cancer with respect to queuosine. AB - Queuosine is a highly modified nucleoside analogue of guanosine. It is present only in the first position of anticodon loop of specific tRNA i.e., tRNA(his), tRNA(asp), tRNA(asn) and tRNA(tyr) and post transcriptionally modified with base for-base exchange of guanine to queuine. The transfer RNA modifying enzyme transfer RNA guanine transglycosylase (TGTase) catalyzes the modification of tRNAs. Transfer RNA is completely modified with respect to queuosine in mature tissue, however modification is often incomplete in mitotically active cells. Hypomodification of transfer RNA is correlated with cell proliferation and malignancy. In the present study queuosine modification of transfer RNA and TGTase activity is compared in normal, Dalton's lymphoma ascites transplanted (DLAT) cancerous and queuine treated DLAT cancerous mouse liver. Transfer RNA of cancerous mouse is hypomodified in terms of queuosine modification. TGTase activity of cancerous mouse is found to decrease to less then half of enzyme activity of normal mouse; suggesting that the enzyme may be responsible for transfer RNA hypomodification. Exogenous treatment of queuine during development of cancer improves the queuosine modification of transfer RNA. The activators NaPP and ATP enhance TGTase activity of normal and DLAT cancerous mouse, where as 7mG inhibits the TGTase activity. PMID- 17114932 TI - A cup full of functions. AB - Data from different laboratories have recently indicated that Cup is a multi functional protein acting both during Drosophila ovary development and early embryogenesis. Cup directly and/or indirectly affects the activity of different mRNAs and proteins to achieve a broad range of biological functions: (1) Cup interacts with Nanos to promote maintenance and survival of the female germ-line stem cells; (2) it binds eIF4E and 3'-UTR-bound factors to repress translation of oskar and nanos mRNAs; (3) it interacts genetically with eIF4E to control translation initiation during ovary development and growth; (4) it may play a role in the control of the phosphorylation status of eIF4E within the developing ovary; finally, (5) it possesses nucleo-cytoplasmic shuttling properties thus pointing to a still uncharacterized function in the cell nucleus. A multi disciplinary approach, ranging from genetics to proteomics, will be required to shed light on the diverse molecular mechanisms involving Cup and the growing family of its specific interactors. PMID- 17114933 TI - Polymorphic CUG repeats in human mRNAs and their effects on gene expression. AB - Expanded CUG repeats in the 3'-untranslated region (UTR) of the gene encoding myotonic dystrophy protein kinase (DMPK) cause myotonic dystrophy type 1 disease (DM1). The presence of such repeats has been found to impede gene expression at several levels in model systems. We took a bioinformatic approach to survey all human mRNA sequences for polymorphic CUG repeats. Our survey revealed that CUG repeats occur widely in various regions of mRNAs, with higher frequency in protein coding regions than 5'-UTRs or 3'-UTRs. About 30 genes were found to contain CUG repeats that are polymorphic in the number of repeats, suggesting the potential to expand or shrink. However, long polymorphic repeats were restricted to the 3'-UTR of the DMPK gene and the coding region of the ribosomal protein L14 gene. Using cell-free translation systems, we showed that extended CUG repeats can inhibit protein synthesis in vitro in the rabbit reticulocyte lysate, but not in wheat germ extracts, consistent with our previous finding of an interaction of CUG repeats with the protein kinase PKR. In transfected cells, CUG repeats can inhibit gene expression both in cis and in trans. However, observations with PKR minus cells indicate that these effects are not primarily attributable to the interaction of extended CUG repeats with PKR. Northwestern blotting detected the presence in human cells of more CUG-binding proteins than are currently known. PMID- 17114934 TI - The NYN domains: novel predicted RNAses with a PIN domain-like fold. AB - Using sensitive sequence profile searches and contextual information gleaned from domain architectures and predicted operons we identify a novel family of protein domains with predicted ribonuclease activity. These domains are found in the eukaryotic proteins typified by the Nedd4-binding protein 1 and the bacterial YacP-like proteins (Nedd4-BP1, YacP nucleases; NYN domains). We show that the NYN domain shares a common protein fold with two other previously characterized groups of nucleases, namely the PIN (PilT N-terminal) and FLAP/5' --> 3' exonuclease superfamilies. We also show that all these proteins share a common set of 4 acidic conserved residues that are predicted to constitute their active site. Based on the conservation of the acidic residues and structural elements we suggest that PIN and NYN domains are likely to bind only a single metal ion, unlike the FLAP/5' --> 3' exonuclease superfamily, which binds two metal ions. We also present evidence that the other conserved residues shared by all these three domains are likely to play critical roles in sensing the substrate and positioning the catalytic residues in the right conformation. Based on conserved gene neighborhoods we infer that the bacterial members are likely to be components of the processome/degradsome that process tRNAs or ribosomal RNAs. Eukaryotic versions appear to have undergone extensive functional diversification as suggested by the several distinctive multi-domain architectures showing fusions with various other RNA-binding domains like CCCH, PPR and KH domains. Interestingly, the eukaryotic NYN domains also show multiple fusions to the UBA domain, an ubiquitin-binding adaptor domain. This observation, together with the monoubiquitination of Nedd4-BP1 by the ubiquitin ligase Nedd4 suggests that the NYN domain proteins of eukaryotes are regulated by monoubiquitination. Given the localization of Nedd4-BP1 to punctuate nuclear bodies, it is likely that they are parts of nuclear RNA-processing complexes that are dependent on monoubiquitination for their assembly. PMID- 17114935 TI - The prokaryotic origin of the pathways for synthesis and post-synthetic modification of deoxyribonucleic acid. AB - A previous study showed that in mammals the pathways leading to synthesis and post-synthetic modification of DNA employ methionine as common donor of atoms: the carbon coming from the methyl group of this amino acid is needed for replication; its entire methyl group is needed to build m(5)C on semiconservatively newly replicating chains. This work showed that the two pathways originate in bacteria where an enzymatic system forms, on DNA, m(6)A in addition to m(5)C. The formation rate of m(6)A gradually decreased during the bacterial CGC, while that of m(5)C reached an optimum in its middle. This shift suggested that the dcm and dam methyltransferase activities, as well as the activities of the methyltransferase moieties of the RM enzymes, are uncoupled. PMID- 17114936 TI - Discrimination of non-protein-coding transcripts from protein-coding mRNA. AB - Several recent studies indicate that mammals and other organisms produce large numbers of RNA transcripts that do not correspond to known genes. It has been suggested that these transcripts do not encode proteins, but may instead function as RNAs. However, discrimination of coding and non-coding transcripts is not straightforward, and different laboratories have used different methods, whose ability to perform this discrimination is unclear. In this study, we examine ten bioinformatic methods that assess protein-coding potential and compare their ability and congruency in the discrimination of non-coding from coding sequences, based on four underlying principles: open reading frame size, sequence similarity to known proteins or protein domains, statistical models of protein-coding sequence, and synonymous versus non-synonymous substitution rates. Despite these different approaches, the methods show broad concordance, suggesting that coding and non-coding transcripts can, in general, be reliably discriminated, and that many of the recently discovered extra-genic transcripts are indeed non-coding. Comparison of the methods indicates reasons for unreliable predictions, and approaches to increase confidence further. Conversely and surprisingly, our analyses also provide evidence that as much as approximately 10% of entries in the manually curated protein database Swiss-Prot are erroneous translations of actually non-coding transcripts. PMID- 17114937 TI - Evidence for control of splicing by alternative RNA secondary structures in Dipteran homothorax pre-mRNA. AB - In a recent study that identified highly evolutionary conserved sequences in three genomes of Diptera species we described an ultraconserved element found at an internal exon-intron junction of the Drosophila melanogaster homothorax (hth) gene that appeared to be involved in the control of hth pre-mRNA splicing. We also discussed a possible role of RNA secondary structure at this site in the regulation of hth pre-mRNA splicing. In this report we identify a shorter evolutionary conserved intronic element within the hth gene that is located downstream of the first element and has sequence complementarity to it. We demonstrate that intramolecular interactions between these two elements would give rise to alternative RNA secondary structures, which in turn may result in differential control of homothorax pre-mRNA splicing. We also provide additional comparative genomic data from several newly available insect genomes supporting our original conclusion that these conserved elements are important in the post transcriptional regulation of homothorax gene expression in Diptera. PMID- 17114939 TI - The cap-dependent translation apparatus integrates and amplifies cancer pathways. AB - Deregulation of a plethora of cancer genes causes pathological changes in only a small set of pathways that confer a cell with malignant properties. This principle of convergence of oncogenic signaling-the ability of several hundred oncogenes to focus their effects on a few critical regulatory nodes that impart autonomy to the cell-motivates the search for putative focal points. Genomic, transcriptional and posttranslational mechanisms that regulate the function of cancer gene pathways are all well established. It has recently become evident that cancer is also subject to translational control. Here we discuss cancer related regulatory events that are mediated by the cap-dependent mRNA binding stage of translation initiation. This information implicates the cap-dependent protein synthesis pathway as a pleotropic integrator and amplifier of many essential oncogenic signals, and the translational control network as a bona fide molecular target for anti-cancer therapeutic interventions. PMID- 17114938 TI - A-to-I RNA editing and human disease. AB - The post-transcriptional modification of mammalian transcripts by A-to-I RNA editing has been recognized as an important mechanism for the generation of molecular diversity and also regulates protein function through recoding of genomic information. As the molecular players of editing are characterized and an increasing number of genes become identified that are subject to A-to-I modification, the potential impact of editing on the etiology or progression of human diseases is realized. Here we review the recent knowledge on where disturbances in A-to-I RNA editing have been correlated with human disease phenotypes. PMID- 17114940 TI - Altered expressions of the noncoding hsromega gene enhances poly-Q-induced neurotoxicity in Drosophila. AB - In an earlier report two P-transposon insertion alleles of the noncoding hsromega gene, hsromega(05241) and P292 were shown to enhance neurodegeneration caused by expression of ataxin-1 protein with expanded poly-Q in a Drosophila model. In present study, we examined the possible relation between hsromega gene expression and toxicity due to poly-Q pathogenesis. The Drosophila hsromega gene produces several noncoding transcripts in almost all cell types, of which the >10 kb long hsromega-n transcript organizes heterogeneous RNA binding (hnRNPs) and related proteins as nucleoplasmic omega speckles. We show that P insertion alleles of the hsromega gene, which cause its overexpression, dominantly enhance neurodegeneration in fly eyes expressing either expanded poly-Q (127Q) or mutant huntingtin protein. Null allele of Hrb87F gene, encoding hnRNPA1, and a novel gene's mutant allele (l(3)pl10(R)), which affects the omega speckles, also dominantly enhance 127Q-induced neurodegeneration. The hsromega-n transcripts or the hnRNPs do not colocalize with the poly-Q nuclear inclusion bodies, neither in hsromega wild type, nor in hsromega mutant background. However, the levels of poly-Q and Hsp70 were significantly higher in hsromega mutant eye discs. Sequestration of hnRNPs and other related RNA-binding proteins by overexpression of hsromega transcripts in hsromega(05241) or in l(3)pl10(R) background or the reduced levels of Hrb87F protein seem to affect nuclear RNA metabolism, thus enhancing the toxicity due to poly-Q expansion. PMID- 17114941 TI - An apical GAGA loop within 5' UTR of the coxsackievirus B3 RNA maintains structural organization of the IRES element required for efficient ribosome entry. AB - Coxsackievirus B3 (CVB3) a close relative of Poliovirus has been implicated as the primary cause of dilated cardiomyopathy in humans. The single-stranded, plus sense, viral genomic RNA is naturally uncapped and the translation of viral encoded protein is initiated from an 'internal ribosome entry site' (IRES) element at the 5' untranslated region (UTR). The 741 nucleotide 5'UTR is highly structured, with defined stem loops. Previously, we have reported results of probing the secondary structure of the 5'UTR RNA, and showed the existence of 11 putative stem loop (SL) structures, SL A-K. Further, using mutants, we showed the importance of an apical GAGA loop in SL-H, in IRES activity. In addition to significantly reducing CVB3 IRES activity the mutations in the 5'UTR drastically reduced interaction with La autoantigen, a critical trans acting factor. Here, we demonstrate that mutation of the GAGA loop to CAGU alters the structure around SL H, and reduces the interaction of La protein with critical cis acting elements of the IRES RNA. Interestingly, mutation of the loop to GAAA also has similar effect, suggesting the GAGA sequence of the loop, is necessary for the function. Furthermore, using gene specific siRNA against La protein we have demonstrated La protein to be a bona fide trans acting factor for CVB3 IRES. However, upon partial knockdown of La protein, translation mediated by the mutant 5'UTRs are affected to a similar extent as the wild type, implicating the structural alteration as the reason for the effect of the mutations on IRES activity. PMID- 17114942 TI - Development and application of a high-throughput assay for glmS riboswitch activators. AB - Riboswitches are newly-discovered gene control elements that are promising targets for antibacterial drug development. To facilitate the rapid discovery and development of riboswitch-targeted compounds, modern drug discovery techniques such as structure-based design and high-throughput screening will need to be applied. One promising riboswitch drug target is the glmS riboswitch, which upon binding glucosamine-6-phosphate (GlcN6P) functions as a ribozyme and catalyzes self-cleavage. Herein we report the development of a high-throughput assay for glmS ribozyme cleavage that relies on fluorescence resonance energy transfer (FRET). This assay can be used to screen for compounds that bind to and activate glmS ribozyme cleavage. To validate the screen, we demonstrate that the assay can identify the active compounds from a library of GlcN6P analogs whose affinities for ribozyme were determined by commonly used electrophoretic methods with radiolabeled RNA. Furthermore, the primary screen of a library of 960 compounds previously approved for use in humans identified five active compounds, one of which is a GlcN6P analog known to stimulate ribozyme activity. These results demonstrate that modern high-throughput screening techniques can be applied to the discovery of riboswitch-targeted drug compounds. PMID- 17114943 TI - MicroRNA modulation of RNA-binding protein regulatory elements. AB - We propose that microRNAs could modulate RNA-binding protein binding sites in a dynamic manner. We suggest that the cis-regulatory code targeted by microRNAs is, at least in part, the same as that read by mRNA-binding proteins. Our hypothesis predicts that microRNAs indirectly or directly bind to RNA-binding protein binding sites. Alternatively, the microRNA-mRNA interactions can themselves be the target of an mRNA-binding protein. Lastly, we envision examples where multiple mRNA regulatory elements are simultaneously influenced by microRNA-mRNA interactions such that the binding of one or more microRNA results in conformational changes in the structure of the mRNA, thereby, either revealing or masking a second regulatory element. PMID- 17114944 TI - Gene silencing with siRNA duplexes composed of target-mRNA-complementary and partially palindromic or partially complementary single-stranded siRNAs. AB - Synthetic small interfering RNA (siRNA) duplexes are widely used to transiently and sequence-specifically disrupt gene expression in mammalian cultured cells. The efficiency and specificity of mRNA cleavage is partly affected by the presence of the nontargeting "passenger" or "sense" siRNA strand, which is required for presentation of the target-complementary or guide siRNA strand to the double-strand-specific RNA silencing protein machinery. We show that siRNA duplexes can be designed that are solely composed of two fully target complementary guide strands that are sufficiently complementary to each other to form stable duplexes with characteristic 3' overhanging ends. The general feasibility of this approach is documented by transient knockdown of lamin A/C and emerin in HeLa cells. The silencing efficiencies of guide-only siRNA duplexes are comparable to prototypical fully paired passenger/guide duplex siRNAs, even though guide-only siRNA duplexes may contain a significant number of nonWatson Crick and G/U wobble base pairs. Such siRNA duplexes may offer advantages regarding production costs and specificity of gene silencing. PMID- 17114945 TI - The splicing factor PSF is part of a large complex that assembles in the absence of pre-mRNA and contains all five snRNPs. AB - PSF (PTB-associated splicing factor) is a large nuclear protein that has been implicated in numerous processes including transcription and RNA splicing. It has been shown to directly associate with U5 snRNA and has also been found within numerous purified splicing complexes. Here, we show that when HeLa nuclear extracts are adjusted to splicing conditions, PSF is found as part of a large complex that contains all five snRNPs and most known splicing factors. Formation of the complex does not require addition of exogenous pre-mRNA substrate and occurs at 4 degrees C but is salt sensitive. Sedimentation experiments and identification of individual components by mass spectrometry revealed association with multiple nuclear factors, most of which overlap with spliceosome components. PMID- 17114946 TI - A non-discriminating aspartyl-tRNA synthetase from Halobacterium salinarum. AB - The tRNA-dependent transamidation pathway is the essential route for Asn tRNA(Asn) formation in organisms that lack an asparaginyl-tRNA synthetase. This pathway relies on a nondiscriminating aspartyl-tRNA synthetase (ND-AspRS encoded by aspS), an enzyme with relaxed tRNA specificity, to form Asp-tRNA(Asn). The misacylated tRNA is then converted to Asn-tRNA(Asn) by the action of an Asp tRNA(Asn) amidotransferase. Here we show that Asn-tRNA(Asn) formation in the extreme halophile Halobacterium salinarum also occurs by this transamidation mechanism, and we explore the property of the haloarchaeal AspRS to aspartylate tRNA(Asn) in vivo and in vitro. Transformation of the E. coli trpA34 strain with the H. salinarum aspS and tRNA(Asn) genes led to restoration of tryptophan prototrophy by missense suppression of the trpA34 mutant with heterologously in vivo formed Asp-tRNA(Asn). The haloarchaeal AspRS works well at low and high (0.1 3 M) salt concentrations but it is unable to use Escherichia coli tRNA as substrate. We show that mutations of two amino acids (H26 and P84) located in the AspRS anticodon binding domain limit the specificity of this nondiscriminating enzyme towards tRNA(Asn). Thus, as was observed in an archaeal discriminating AspRS and a bacterial ND-AspRS, amino acids in these positions influence the enzyme's tRNA selection. PMID- 17114947 TI - Crystal structure of tRNA pseudouridine synthase TruA from Thermus thermophilus HB8. AB - The pseudouridine synthase (Psi synthase) TruA catalyzes the conversion of uridine to pseudouridine at positions 38, 39 and/or 40 in the anticodon stem-loop (ASL) of tRNA. We have determined the crystal structure of TruA from Thermus thermophilus HB8 at 2.25 A resolution. TruA and the other (Psi synthases have a completely conserved active site aspartate, which suggests that the members of this enzyme family share a common catalytic mechanism. The T. thermophilus TruA structure reveals the remarkably flexible structural features in the tRNA-binding cleft, which may be responsible for the primary tRNA interaction. In addition, the charged residues occupying the intermediate positions in the cleft may lead the tRNA to the active site for catalysis. Based on the TruB-tRNA complex structure, the T. thermophilus TruA structure reveals that the tRNA probably makes the melting base pairs move into the cleft, and suggests that a conformational change of the substrate tRNA is necessary to facilitate access to the active site aspartate residue, deep within the cleft. PMID- 17114948 TI - Post-transcription meets post-genomic: the saga of RNA binding proteins in a new era. AB - Gene expression is regulated by a complex series of events that take place both at the transcriptional and post-transcriptional levels. The mechanisms and factors involved in transcriptional regulation are relatively well understood, whereas post-transcriptional regulation, in comparison, is still a poorly appreciated process. RNA binding proteins (RBPs) are the key regulators of all post-transcriptional events (RNA splicing, stability, transport and translation). Essentially, in order to improve our knowledge in post-transcriptional regulation, we need to elucidate the mechanisms employed by RBPs to control gene expression. The combination of genomic tools with traditional biochemical approaches generated novel technologies, like ribonomics. The application of these novel technologies not only had a profound impact in the study of RBPs but also created the ground work necessary for the identification of post transcriptional gene networks; a selected group of mRNAs associated with a particular function or biological pathway/ process that is regulated by the same RBP. In this review article, we explore the latest achievements and discuss future challenges that lie ahead in the RBP world. PMID- 17114950 TI - Trends in the adoption of medications for alcohol dependence. AB - Increasing attention is being paid to the development and dissemination of effective pharmacotherapies for the treatment of alcohol and other drug dependence. However, numerous structural and philosophical barriers impede the widespread adoption of these treatment approaches in everyday clinical practice. Research is needed to understand and overcome this gap. Drawing upon data collected from 2 large samples of substance abuse treatment providers at multiple time points, this article examines the prevalence and correlates of the adoption of the currently available pharmacotherapies for alcohol dependence: disulfiram, oral naltrexone, and acamprosate. These data suggest that the proportion of treatment programs using pharmacotherapies for alcohol dependence has been declining over time. In addition, the proportion of patients to whom these medications are prescribed is notably low. The adoption of disulfiram and naltrexone is significantly more likely in programs that are accredited, employ at least 1 physician, offer integrated care for patients with co-occurring psychiatric conditions, derive proportionately more revenue from commercial insurance payers, and have fewer linkages with the criminal justice system. Preliminary data suggest that the early adoption of acamprosate is following a similar pattern. Recommendations for addressing challenges to the diffusion of pharmacotherapies for alcohol dependence are presented. PMID- 17114952 TI - A rational approach to the pharmacotherapy of alcohol dependence. AB - The use of alcohol is widespread in the world, and although there are many regular users, some individuals drink excessively. Understanding the time course of the development and maintenance of alcohol dependence is important in assessing the potential risk/benefit of an intervention and in accurately treating the problem. Most, although not all, literature suggests that alcoholism is a chronic, relapsing disorder and that there is a general progression from less to more severe problems. Understanding the neurobiology that underlies alcohol dependence as it relates to different clinical stages may help in the development of effective targeted pharmacological treatments. Important clinical stages that may be amenable to pharmacological intervention include the transition from alcohol use to heavy drinking particularly in vulnerable individuals, the cessation of heavy drinking in individuals who want to quit, and the prevention of relapse in individuals who have initiated abstinence but may struggle with craving or the desire to resume alcohol use. Neurotransmitter systems implicated in these stages include glutamate, gamma-aminobutyric acid, opioid, and serotonin systems that may act directly or via the indirect modulation of dopamine function. The treatment implications will also be discussed. PMID- 17114954 TI - Combining psychosocial treatment with pharmacotherapy for alcohol dependence. AB - Pharmacotherapy for alcohol dependence is always delivered in a psychosocial context that may affect the outcome of the treatment. The rigorous study of different psychotherapeutic treatments for alcohol dependence has shown several distinct approaches to be effective. This article reviews the combination of alcohol dependence pharmacotherapies, including disulfiram, naltrexone, and acamprosate, with different psychosocial interventions. Many psychosocial interventions for alcohol dependence, including Alcoholics Anonymous, can be integrated successfully with pharmacotherapy. Psychosocial interventions, ranging from brief medical management to more intensive manualized psychotherapies, have all been shown to produce positive outcomes in certain studies, depending on the specific medication and the study context. Particularly successful combinations may include the use of behavioral marital therapy plus a disulfiram contract for patients taking that medication, and the combination of naltrexone or acamprosate with cognitive-behavioral therapy or psychosocial support. Ongoing research examining the optimal combinations of medications with different psychosocial treatments for alcohol dependence may further inform the field. PMID- 17114955 TI - Medulloblastoma simulating acute myeloid leukemia: case report with a review of "myeloid antigen" expression in nonhematopoietic tissues and tumors. AB - Medulloblastoma is a primitive neuroectodermal tumor arising in the posterior fossa usually in the first decade of life. Systemic metastases are infrequent at diagnosis and usually occur after surgical resection or shunt placement. We report a rare case of medulloblastoma in an 18-year-old woman who presented with headache, leukopenia, and anemia. Neurologic examination was normal. Bone marrow evaluation revealed primitive cells morphologically resembling blasts. By flow cytometry, these cells lacked CD45 and expressed CD13/33, CD15, CD34, HLA-DR, and strong CD56. The presence of myeloid antigens and CD34 suggested acute myeloid leukemia; however, the bone marrow core biopsy architecture and tumor cells in cerebrospinal fluid were more compatible with a nonhematopoietic tumor. Further workup revealed a cerebellar mass, and a diagnosis of desmoplastic medulloblastoma was made. To our knowledge, this is the first reported case of a nonhematopoietic small round blue-cell tumor expressing multiple myeloid antigens and CD34 by flow cytometry. PMID- 17114956 TI - Routine use of PET scans after completion of therapy in pediatric Hodgkin disease results in a high false positive rate. AB - PURPOSE: Fluoro-2-deoxy-D-glucose positron emission tomography scans are becoming standard of care in the evaluation of Hodgkin disease (HD). The frequency of false positive (FP) PET scans in pediatric HD after completion of therapy has not been well studied. PATIENTS AND METHODS: All PET scan reports on pediatric HD patients at our institution between February 2000 and February 2005 were retrospectively reviewed. Scans were considered positive if the interpretation was most consistent with malignancy. FP results were determined by pathologic evaluation, resolution on scan, or absence of disease progression over at least 1 year without intervention. RESULTS: We reviewed 255 PET scans on 47 patients, including 156 posttherapy scans on 34 patients. Positive predictive value for scans obtained during routine follow-up was 11%, with an FP rate of 16%. Identifiable etiologies of FP scans included: fibrosis, progressive transformation of germinal centers, abdominal wall hernia, appendicitis, thymus and HIV associated lymphadenopathy. CONCLUSIONS: Routine PET scans after completion of therapy in pediatric HD patients have a low positive predictive value and a high FP rate. Prospective studies are needed to reduce the ambiguity of positive results. In the interim, positive PET scans after treatment should be interpreted cautiously and therapeutic decisions should not be made without histologic confirmation. PMID- 17114957 TI - Imerslund-Grasbeck syndrome associated with recurrent aphthous stomatitis and defective neutrophil function. AB - Vitamin B(12) deficiency is a well-known cause of recurrent aphthous stomatitis (RAS). However, the mechanism by which this deficiency causes the stomatitis is not well understood. Imerslund-Grasbeck syndrome (IGS) causes vitamin B(12) deficiency and proteinuria due to a defect in the vitamin B(12) receptor. We sought to determine whether the RAS observed in IGS patients is associated with neutrophil dysfunction. We report 3 infants with vitamin B(12) deficiency due to IGS, who presented with borderline or normal hemoglobin concentrations, RAS, and a neutrophil function defect. All 3 patients were homozygous for a splice site mutation affecting exon 4 of the AMN gene. A direct correlation was observed between low serum vitamin B12 levels and defective neutrophil function (low chemotaxis and elevated superoxide production) in the patients. Vitamin B(12) therapy led to an immediate resolution of aphthous stomatitis and full correction of neutrophil function. We demonstrated that serum vitamin B(12) deficiency is associated with a neutrophil chemotactic defect and RAS in IGS patients. We suggest that the RAS observed in these patients is due to this defect. PMID- 17114958 TI - A pilot pharmacokinetic and antiangiogenic biomarker study of celecoxib and low dose metronomic vinblastine or cyclophosphamide in pediatric recurrent solid tumors. AB - Tumor vasculature is a reasonable target for cancer therapy and lower more frequent doses of traditional chemotherapeutics [low-dose metronomic (LDM) chemotherapy] has been shown to have antiangiogenic efficacy. This study evaluated the safety and pharmacokinetics of celecoxib and LDM vinblastine or cyclophosphamide in children with recurrent, refractory solid tumors. We also investigated whether a subset of circulating plasma proteins are surrogate markers of angiogenic activity. Thirty-three children were enrolled in this pilot study and received celecoxib (250 mg/m(2) PO b.i.d.) and either vinblastine (1 mg/m(2) IV 3 x /wk) or cyclophosphamide (30 mg/m(2) PO daily) continually. Celecoxib alone and with LDM chemotherapy was well tolerated and plasma concentrations were consistent with those shown to have antiangiogenic activity. Four patients (13%) had durable stable disease (28 to 78 wk) although no complete or partial responses were observed. The surrogate markers measured (vascular endothelial growth factor, basic fibroblast growth factor, soluble vascular cell adhesion molecule, soluble intercellular cell adhesion molecule, endostatin, and thrombospondin-1) were highly variable and no statistically significant relationship between them and disease progression or maintenance of stable disease was observed. We concluded that this regimen is well tolerated hence supporting the use of this form of therapy in pediatric patients. However, future studies should include more homogenous patient populations and focus on validating surrogate markers to monitor treatment activity. PMID- 17114959 TI - Teaching teachers about childhood cancer: the effects of a web-based training program. AB - The purpose of this study was to examine the utility and acceptability of a modular computer-based training program on childhood cancer (eg, acute and late effects of treatment, intervention strategies) for teachers. A within-subjects design was implemented with 41 teachers and teachers in training. Participants completed tests of childhood cancer knowledge and application skills both before and after completing the web-based training. An acceptability questionnaire was completed after the training. Results indicated significant gains in knowledge and in case application, as well as high levels of acceptability of the training. It seems that a web-based training program can be accessed by teachers and in 2 to 4 hours can significantly increase cancer knowledge in an acceptable manner. PMID- 17114960 TI - Preleukemic TEL-AML1-positive clones at cell level of 10(-3) to 10(-4) do not persist into adulthood. AB - The TEL-AML1 translocation, t(12;21)(p13;q22), is one of the most frequent genetic aberrations in childhood B-cell precursor acute lymphoblastic leukemia (ALL), where it occurs in 25% of all cases. In contrast, the translocation is seen in only 3% of adult ALL cases. Evidence suggests that the TEL-AML1 translocation occurs in utero in 1% of all newborn children at cell levels of 10 to 10. In this study, we explore the prevalence of TEL-AML1-positive cells in 2 cohorts of healthy blood donors by real-time and nested reverse transcription polymerase chain reaction. Overall, TEL-AML1-positive cells were demonstrated in 10 of 2005 healthy donors, that is, a prevalence of 0.5% (95% confidence interval, 0.2-0.3%). The level of TEL-AML1-positive cells was estimated to 10 to 10. The observed prevalence of TEL-AML1-positive cells in healthy adults is of the same order of magnitude as the prevalence reported in healthy newborns, but the observed cell level of 10 to 10 is much lower. These data indicates that prenatal TEL-AML1 subclones does not persist throughout adult life at cell levels of 10 to 10. The findings are compatible with the risk of t(12;21)(p13;q22) ALL correlating with the total number of TEL-AML1-positive cells in peripheral blood in both childhood and adulthood. PMID- 17114961 TI - The first infant case with hepatosplenic gammadelta T-cell lymphoma after acute disseminated encephalomyelitis (ADEM)-like exacerbation. AB - We report the first infant case with hepatosplenic gammadelta T-cell lymphoma after recurrent acute disseminated encephalomyelitis-like, which were rapidly resolved with steroid pulse therapy. The patient had a history of recurrent bronchitis, intractable diarrhea, and failure to thrive since 4 months of age. Immunologic analysis revealed higher percentage of circulating gammadelta T-cells with markedly reduced numbers of CD3TCRalphabetaCD8 T-cells. The patient developed gammadelta T-cell lymphoma at the age of 15 months. Clinical course of the patient suggests the importance of immunological background for the development of hepatosplenic gammadelta T-cell lymphoma. PMID- 17114962 TI - A case report of langerhans histiocytosis presenting sequentially over a 21-year period with Letterer-Siwe disease, hand-Schuller-Christian disease and eosinophillic granuloma of bone. AB - Langerhans cell histiocytosis (LCH) is a very rare disorder, and usually considered a disease of childhood. The adult form of LCH is even rare, and in some case may be considered to represent delayed presentation of a pathologic process beginning in childhood. This case report highlights the range of presentations of LCH possible even in a single patient, and supports the contention that some adult LCH cases may be due to delayed presentation of childhood LCH. PMID- 17114963 TI - Germ cell tumor showing partial trisomy 1 in a gonadectomized intersex child with monosomy X and double Y mosaicism. AB - High incidence of germ cell tumors arising from dysgenetic gonads in patients with sexual chromosome abnormalities has been described, especially in patients with a Y chromosome bearing cell line. Here we report a 14-year-old patient with ambiguous genitalia. Constitutional karyotype showed 45,X/46,X,derY [?t(Yp;Yq)] mosaicism. The patient developed an abdominally located mixed malignant germ cell tumor 5 years after the removal of the dysgenetic gonads. Tumor karyotype showed partial trisomy 1q, a derivative 8q, and a hyperdiploidy with +X, +7, +12, +15, +19, +21, and an unidentified marker. PMID- 17114964 TI - An extremely uncommon complication of ITP: spontaneous rupture of an ovarian follicle cyst and massive intra-abdominal bleeding. AB - Idiopathic thrombocytopenic purpura usually presents with minor bleeding such as petechia and purpura. Rarely, life-threatening events as intracranial and intra abdominal bleedings can be seen. We would like to present a rare case diagnosed as idiopathic thrombocytopenic purpura, presenting with abdominal pain and paleness. In this 17-year-old female patient, extensive abdominal sensitivity was revealed on physical examination and massive intra-abdominal hemorrhage secondary to distended ovarian follicle rupture was seen on ultrasonography and abdominal computed tomography. The case was treated successfully with intravenous immunoglobin, thrombocyte suspension, and pulse methylprednisolone. PMID- 17114965 TI - Concomitant Candida epiglottitis and disseminated Varicella zoster virus infection associated with acute lymphoblastic leukemia. AB - Acute epiglottitis by nonbacterial pathogens is an uncommon but life-threatening clinical entity. Herein, we report the concomitant occurrence of Candida epiglottitis and mucosal and visceral Varicella zoster virus infection in a child with acute lymphoblastic leukemia. Both infections were atypical in their presentation, occurred in a severely immunocompromised host, and required invasive procedures for diagnosis. PMID- 17114966 TI - Clinical manifestations of hemoglobin Chico at high altitude. AB - Hemoglobin Chico is a rare hemoglobinopathy characterized by low oxygen affinity and a right-shifted oxygen dissociation curve. Detailed clinical evaluations of affected individuals have not been previously reported. We therefore report on the clinical features of Hemoglobin Chico in a Latino male living at high altitude, who desired to participate in school sports. As a young boy with asthma, he had the unusual finding of growth delay and digital clubbing which improved with asthma control. At 16 years of age, he had mild anemia and a decreased pulse oximetry (83%) but sufficient pulmonary reserve to participate in physically demanding activities. PMID- 17114967 TI - Donor cell-derived acute myeloblastic leukemia after allogeneic peripheral blood hematopoietic stem cell transplantation for juvenile myelomonocytic leukemia. AB - Despite its rarity, donor cell leukemia (DCL) is a most intriguing entity. We report here the case of a 5 year-old girl with juvenile myelomonocytic leukemia and normal female karyotype who developed acute myeloblastic leukemia with a karyotype of 46, X, t(X; 7) (p21; p11.2), der(7) t(3; 7) (q13.3; q22) 5 months after peripheral blood hematopoietic stem cell transplantation from her HLA matched sister. We performed the analysis of short tandem repeat sequence markers to DNA obtained from donor peripheral blood, patient's peripheral blood including leukemic blasts and patient's hair root. This analysis showed that the leukemic blood DNA matched the donor blood DNA and not the patient's DNA, thus confirming DCL. To our knowledge, this is the first case of DCL after peripheral blood SCT for juvenile myelomonocytic leukemia. PMID- 17114968 TI - Endodermal sinus tumor of the infant vagina treated exclusively with chemotherapy. AB - Endodermal sinus tumor (EST) of the vagina is a rare malignancy usually diagnosed before 3 years of age. Historically, the approach to therapy has included radical surgical resection, with adjuvant irradiation, and chemotherapy. An infant presented with vaginal bleeding, imaging evidence of a vaginal mass and an elevated alpha-fetoprotein level. Examination under anesthesia with vaginal biopsies confirmed the diagnosis of an EST (yolk sac) tumor of the vagina. After 5 cycles of chemotherapy, the alpha-fetoprotein had normalized and repeat vaginal biopsies for suspected residual disease was negative for malignancy. To allow preservation of sexual and reproductive function, chemotherapy as a sole modality of treatment for EST should be considered. PMID- 17114969 TI - Autologous hematopoietic stem cell transplantation in children with relapse or refractory Hodgkin disease. PMID- 17114970 TI - Eighth Wolf Creek Conference on Cardiopulmonary Resuscitation: addressing the scientific basis of reanimation. Preface. PMID- 17114971 TI - Definition of successful defibrillation. AB - OBJECTIVES: The definition of defibrillation shock "success" endorsed by the International Liaison Committee on Resuscitation since the publication of Guidelines 2000 for Cardiopulmonary Resuscitation and Emergency Cardiac Care has been removal of ventricular fibrillation at 5 secs after shock delivery. Although this success criterion provides a direct assessment of the primary task of a shock, it may not be the only clinically useful measure of shock outcome. We evaluated a different defibrillation success criterion to determine whether it could provide additional insight into the relative performance of different defibrillation shocks. DESIGN: A randomized study comparing monophasic and biphasic waveform shocks is reported with return of organized rhythm as the primary outcome measure of defibrillation success. PATIENTS: A total of 120 patients with out-of-hospital ventricular fibrillation as the first recorded rhythm were treated with defibrillation with automated external defibrillators. MEASUREMENTS AND MAIN RESULTS: Return of organized rhythm (two QRS complexes, <5 secs apart, <60 secs after defibrillation) was achieved in 31 monophasic shock (45%) and 35 biphasic shock (69%) patients (relative risk, 1.53, 95% confidence interval, 1.11-2.10). Logistic regression analysis revealed that shock waveform was the strongest independent predictor of return of organized rhythm (odds ratio, 4.0; 95% confidence interval, 1.67-10.0). Defibrillation success with the conventional International Liaison Committee on Resuscitation criterion was very high (91% and 98%, respectively) and not significantly different between groups. CONCLUSIONS: Return of organized rhythm proved to be a more sensitive measure of relative defibrillation shock performance than the conventional shock success criterion. Inclusion of return of organized rhythm as an end point in future clinical research could help discern more subtle defibrillation shock effects and contribute to further optimization of defibrillation technology. PMID- 17114972 TI - Difficulty of cardiac arrest rhythm identification does not correlate with length of chest compression pause before defibrillation. AB - OBJECTIVE: Performing high-quality cardiopulmonary resuscitation immediately before electrical defibrillation serves as an important predictor of shock success. Long preshock pauses in cardiopulmonary resuscitation frequently occur, as noted by recent clinical investigations. We sought to determine whether these long pauses were due to difficulties in identifying shockable rhythms or rather due to local factors during resuscitation attempts. DESIGN: Prospective in hospital study of cardiac arrest resuscitation attempts coupled with a retrospective review of preshock pause rhythms by 12 trained providers. Reviewers scored rhythms by ease of identification using a discrete Likert scale from 1 (most difficult to identify) to 5 (easiest to identify). The resuscitation cohort was organized into preshock pause-duration quartiles for statistical analysis. Resident physicians were then surveyed regarding human factors affecting preshock pauses. RESULTS: A total of 118 preshock pauses from 45 resuscitation episodes were collected. When evaluated by quartiles of preshock pause duration, difficulty of rhythm identification did not correlate with increasing pause time. In fact, the opposite was found (longest preshock pause quartile of 23.8-60.2 secs vs. shortest pause quartile of 1.1-7.9 secs; rhythm difficulty scores, 3.2 vs. 3.0; p = .20). When 29 resident physicians who recently served on resuscitation teams were surveyed, 18 of 29 (62.1%) attributed long pauses to lack of time sense during resuscitation, and 16 of 29 (55.2%) thought that room crowding prevented rapid defibrillation. CONCLUSIONS: Long cardiopulmonary resuscitation pauses before defibrillation are likely due to human factors during the resuscitation and not due to inherent difficulties with rhythm identification. This preliminary work highlights the need for more research and training in the area of team performance and human factors during resuscitation. PMID- 17114973 TI - Analysis of the ventricular fibrillation waveform in refibrillation. AB - OBJECTIVES: Frustrating outcomes are driving investigation of alternative resuscitation protocols. Previous analysis of the ventricular fibrillation (VF) waveform has focused on guiding whether to shock immediately or to delay for delivery of cardiopulmonary resuscitation in the case of presenting VF. The same issues emerge in the case of refibrillation. MEASUREMENTS AND MAIN RESULTS: All cases of witnessed VF cardiac arrest in the Rochester, MN, area in a 9-yr period were analyzed. Rochester rescuers employed an early defibrillation protocol during the study period. A summary measure of the VF waveform before the shock delivered in 35 incidents of refibrillation was compared with the time elapsed from the initial shock, the intervening electrocardiographic rhythm, ambulance response time, and call-to-shock time for prediction of early return of spontaneous circulation and of neurologically intact survival. VF waveform analysis separated patients with good outcomes when treated with early defibrillation of refibrillation from those without good outcomes more clearly than other predictors. CONCLUSIONS: Analysis of VF waveform offers promise for real-time guidance of resuscitation efforts on the basis of individual patient characteristics, in refibrillation and in the initial shock. It has advantages over guidance based on individual or aggregate system response times. PMID- 17114974 TI - Preterminal gasping and effects on the cardiac function. AB - OBJECTIVE: Gasping, also known as agonal respirations, is the terminal pattern that occurs after anoxia or ischemia and is a universal phenomenon in mammals. In this article we review the physiology of gasping, the prevalence and significance of gasping in cardiac arrest, and the effects of gasping on cardiac function. DESIGN: Review relevant human and animal literature on gasping and cardiac function during gasping. RESULTS: Gasping originates in the medullary area of the central nervous system. Gasping is prevalent during cardiac arrest: it occurs in all animals during ventricular fibrillation, in a majority of infants (31 of 32) with sudden infant death syndrome, and in 30-40% of witnessed episodes of cardiac arrest in adults. Animal studies demonstrated that gasping is associated with a decrease in intrathoracic pressure during the inspiratory phase, which promotes venous return and an increase in intrathoracic pressure during the expiratory phase, which favors coronary perfusion. Gasping increases cardiac output and cardiac contractility in immature animals exposed to anoxia. CONCLUSIONS: Gasping is auto-resuscitative in immature mammals and improves the outcome of cardiopulmonary resuscitation in mature mammals. Gasping is associated with important cardiorespiratory changes: improved pulmonary gas exchange, increased venous return to the heart, increased cardiac output, cardiac contractility, aortic pressure, and coronary perfusion pressure. PMID- 17114975 TI - Role of buccal PCO2 in the management of fluid resuscitation during hemorrhagic shock. AB - Arterial pressure is a widely used measurement for estimating the severity of hemorrhagic shock and to guide its management. However, this capability is reduced when very low arterial pressure values cannot be reliably measured by noninvasive methods. Moreover, hypoperfusion may be masked by compensatory hemodynamic changes, and therefore, in the presence of near normal blood pressure, tissue hypoperfusion may progress undetected. Accordingly, hypercarbia is a general phenomenon of perfusion failure, which occurs in coincidence of the onset of hypotension and is promptly reversed with restoration of normal blood flows. Increases in buccal mucosa PCO2 are highly correlated with increases in gastric wall and sublingual mucosa PCO2 and decreases in tissue blood flows during hemorrhagic shock. In both clinical and experimental settings, tissue PCO2 measured in the oral mucosa proved to be a practical and reliable measurement for the diagnosis of circulatory failure states and an indicator of its severity. In contrast to intraarterial pressure, buccal PCO2 discriminated between short- and long-term survival after large-volume blood loss. Buccal PCO2 measurement therefore emerges as a useful predictor for survival and outcome and a useful guide to manage fluid resuscitation during hemorrhagic shock. PMID- 17114976 TI - Comparison of buccal microcirculation between septic and hemorrhagic shock. AB - OBJECTIVE: Microcirculatory perfusion is disturbed in sepsis, and global hemodynamics does not necessarily reflect microcirculatory blood flow. In this study, we investigated the effect of the same level of mean arterial pressure (MAP) or cardiac index on the changes in buccal microcirculation between septic and hemorrhagic shock. DESIGN: Prospective, controlled laboratory study. SETTING: University-affiliated research laboratory. INTERVENTIONS: A total of 20 Sprague Dawley rats were divided into four groups: 1) septic shock induced by cecal ligation and perforation: when MAP decreased to 80 mm Hg, saline was infused at a rate of 25 mL.kg.hr for 2 hrs; 2) both time- and MAP-matched hemorrhagic shock: approximately 30% of total blood volume was withdrawn during the corresponding interval, followed by infusion aiming to restore MAP as required when MAP decreased to 80 mm Hg; 3) both time- and cardiac index-matched hemorrhagic shock: approximately 40% of total blood volume was withdrawn during the corresponding interval until MAP decreased to 50 mm Hg, which generally generated a cardiac index similar to those in septic animals, followed by infusion at the same rate for 2 hrs; and 4) sham control: animals underwent the same procedure except no cecal ligation and perforation, bleeding, and infusion. MEASUREMENTS AND MAIN RESULTS: Buccal microcirculation was visualized with the aid of an orthogonal polarization spectral image device. A semiquantitative score was calculated for vessels of <20 mum, primarily representing the capillaries. Impaired buccal capillary blood flows in septic animals were more severe than those in MAP matched hemorrhagic animals and were similar to those in cardiac index-matched hemorrhagic animals during the hypoperfusion period before infusion. Significantly improved global hemodynamics after resuscitation cannot effectively improve the buccal capillary blood flows in septic animals, in contrast to those in MAP-matched and cardiac index-matched hemorrhagic animals. CONCLUSIONS: Impaired microcirculatory alteration in septic shock is more severe than hemorrhagic shock; microcirculation is relatively independent of improved systemic hemodynamics, in contrast to those in hemorrhagic shock. PMID- 17114977 TI - Microcirculation during cardiac arrest and resuscitation. AB - OBJECTIVE: Direct observations of the microcirculation using orthogonal polarization spectral imaging have attracted attention and revealed that, especially in cardiogenic and distributive shock, there is discordance between the macrocirculation and the microcirculation. We evaluated serial changes and the effects of epinephrine on microcirculatory blood flow in the most severe form of circulatory failure, namely, cardiac arrest. DESIGN AND SETTING: : Controlled laboratory animal study. SUBJECTS AND INTERVENTIONS: A total of 15 pigs were subjected to 5 mins of ventricular fibrillation and 5 mins of precordial compression before electrical defibrillation was attempted. In a subset, six animals received 1 mg of epinephrine after 1 min of precordial compression. MEASUREMENTS AND MAIN RESULTS: Microcirculatory blood flow was visualized in the sublingual mucosa at baseline and 0.5, 1, and 5 mins of ventricular fibrillation, at 1 and 5 mins of precordial compression, and at 1 and 5 mins after return of spontaneous circulation. In addition, coronary perfusion pressure was recorded. Microcirculatory blood flow decreased dramatically in the 0.5 min after the onset of ventricular fibrillation. Precordial compression partially restored microcirculatory blood flow in each animal but to a significantly greater extent in animals that achieved return of spontaneous circulation. These changes were paralleled by similar changes in coronary perfusion pressure. Both variables were highly correlated. Administration of epinephrine resulted in a massive reduction of microcirculatory blood flow that lasted for >/=5 mins. CONCLUSIONS: In this model, microcirculatory blood flow was highly correlated with macrocirculatory hemodynamics, including coronary perfusion pressure in distinction with septic shock. Administration of epinephrine dramatically decreased microcirculatory blood flow. PMID- 17114978 TI - Endothelium and cardiopulmonary resuscitation. AB - The endothelium is a viable target for injury, repair and cellular modulation. Because of its vast extension and active metabolic status of producing mediators for vasomotor tone, coagulation, and inflammation, it is a key target for therapy during ischemia/reperfusion injury. Cardiopulmonary resuscitation is a model of whole-body ischemia/reperfusion injury. It has become apparent that the endothelium participates in a host of responses elicited by ischemia/reperfusion. This review examines the role of the endothelium during and after ischemia/reperfusion and the participation by its mediators and evidence for endothelial involvement during and after cardiopulmonary resuscitation. The strategic location of the endothelium makes it an excellent signal transduction mechanism for a host of disease processes. In addition to biochemical stimuli, mechanical stimulation of the endothelium elicits production of several mediators, including endothelium-derived nitric oxide, prostaglandins, and antithrombotics and anticoagulants. Whole-body, periodic acceleration is a novel method of stimulating the endothelium via pulsatile shear stress. Periodic acceleration has been shown to be an effective experimental method of cardiopulmonary resuscitation, with evidence of postresuscitation cardioprotective effects. This review indicates that understanding endothelial modulation during and after ischemia/reperfusion will significantly improve therapeutic choices. PMID- 17114979 TI - Vital organ blood flow with the impedance threshold device. AB - OBJECTIVE: The purpose of this study is to review cardiopulmonary resuscitation hemodynamics and vital organ blood flow in animal models with the use of the impedance threshold device (ITD) and to correlate these findings with the results of human clinical trials. RESULTS: Animal studies have demonstrated near normalization of cerebral blood flow and an increase between 50% and 100% in cardiac blood flow with use of the ITD. Coincident coronary perfusion pressure is significantly increased with the ITD. Results of human clinical trials generally reflect the data seen in animal models, with near normal blood pressure during active compression-decompression cardiopulmonary resuscitation and the ITD, near doubling of blood pressure with standard cardiopulmonary resuscitation plus the ITD, and significantly increased short-term survival rates. CONCLUSIONS: Improved vital organ perfusion with ITD use during cardiopulmonary resuscitation is an important advance in resuscitation. Incorporation of the ITD into protocols that improve other aspects of the care of patients during cardiac arrest and after successful resuscitation should result in further benefit from the ITD. PMID- 17114980 TI - Transient and partial mitochondrial inhibition for the treatment of postresuscitation injury: getting it just right. AB - OBJECTIVE: Within minutes of reperfusing ischemic cardiomyocytes, oxidant stress dramatically increases and is associated with postresuscitation injury. Because mitochondria produce deleterious oxidants and useful metabolic substrates, utilization of electron transport chain inhibitors against reperfusion injury, though promising, must not overly compromise recovery of mitochondrial function. This study sought to further characterize the oxidant source at reperfusion and develop a strategy for therapeutic intervention by manipulation of dose, duration, and the degree of reversibility of mitochondrial inhibition. DESIGN: Comparative laboratory investigation. SETTING: Laboratory of a research university. SUBJECTS: Embryonic chick cardiomyocytes. INTERVENTIONS: Synchronously contracting chick cardiomyocytes were exposed to 1 hr of simulated ischemia and 3 hrs of reperfusion and were monitored for cell viability (propidium iodide) and oxidant generation (dichlorofluorescein). Inhibitors were administered either all course or for the first 15 mins of reperfusion. MEASUREMENTS AND MAIN RESULTS: : Application of diethyldithiocarbamic acid, 2 anthracene-carboxylic acid (rhein tech), and alpha-nicotinamide adenine dinucleotide dehydrogenase (NADH) demonstrated attenuation of the oxidant burst. In addition, diethyldithiocarbamic acid (1 mM), rhein tech (0.1 microM), and alpha-NADH (20 microM) significantly attenuated cell death from a control of 49.7% +/- 6.7% to 15.7% +/- 4.7% (n = 5, p < .01), 26.1% +/- 4.1% (n = 5, p < .01), and 13.8% +/- 1.3% (n = 5, p < .001), respectively. All doses of stigmatellin attenuated reactive oxygen species, but only a 2-20 nM dose during the first 15 mins of reperfusion abrogated cell death from 53.8% +/- 3.5% to 10.8% +/- 2.9% (n = 5, p < .001). Increased doses and durations of stigmatellin abolished reactive oxygen species but augmented injury. Although rotenone (5 microM) attenuated reactive oxygen species, no dose or duration of exposure that ameliorated cell death was found. CONCLUSIONS: Early events of reperfusion are marked by rapid mitochondrial oxidant generation and postresuscitation injury. Electron transport chain blockade provides an effective method of attenuating reactive oxygen species. However, inhibitor administration should be both transient and reversible to necessitate cardioprotection and successful metabolic recovery. PMID- 17114981 TI - Simultaneous blockade of alpha1- and beta-actions of epinephrine during cardiopulmonary resuscitation. AB - OBJECTIVE: Experimental and clinical studies have implicated that alpha1- and beta-adrenergic effects of epinephrine significantly increased the severity of postresuscitation myocardial dysfunction by increasing myocardial oxygen consumption during ventricular fibrillation. This prompted experimental studies to investigate the effect of simultaneous blockade of alpha1- and beta-actions of epinephrine during cardiopulmonary resuscitation. DESIGN: Literature review. RESULTS: Improved postresuscitation myocardial dysfunction was observed in epinephrine-treated animals after its alpha1- and beta-actions were blocked, which were associated with less postresuscitation arrhythmia, lower blood lactate level, better neurologic recovery, and longer duration of survival. CONCLUSIONS: After simultaneous alpha1- and beta-adrenergic blockade, epinephrine administered during cardiopulmonary resuscitation yielded improved postresuscitation myocardial functions and significantly better postresuscitation outcomes. PMID- 17114982 TI - delta-Opioid-induced pharmacologic myocardial hibernation during cardiopulmonary resuscitation. AB - OBJECTIVES: Cardiac arrest and cardiopulmonary resuscitation is an event of global myocardial ischemia and reperfusion, which is associated with severe postresuscitation myocardial dysfunction and fatal outcome. Evidence has demonstrated that mammalian hibernation is triggered by cyclic variation of a delta-opiate-like compound in endogenous serum, during which the myocardial metabolism is dramatically reduced and the myocardium tolerates the stress of ischemia and reperfusion without overt ischemic and reperfusion injury. Previous investigations also proved that the delta-opioid agonist elicited the cardioprotection in a model of regional ischemic intact heart or myocyte. Accordingly, we were prompted to search for an alternative intervention of pharmacologically induced myocardial hibernation that would result in rapid reductions of myocardial metabolism and therefore minimize the myocardial ischemic and reperfusion injury during cardiac arrest and cardiopulmonary resuscitation. DESIGN: Prospective, controlled laboratory study. SETTING: University-affiliated research laboratory. INTERVENTIONS: In the series of studies performed in the established rat and pig model of cardiac arrest and cardiopulmonary resuscitation, the delta-opioid receptor agonist, pentazocine, was administered during ventricular fibrillation. MEASUREMENTS AND MAIN RESULTS: : The myocardial metabolism reflected by the concentration of lactate, or myocardial tissue PCO2 and PO2, is dramatically reduced during cardiac arrest and cardiopulmonary resuscitation. These are associated with less severe postresuscitation myocardial dysfunction and longer duration of postresuscitation survival. CONCLUSIONS: delta-Opioid-induced pharmacologic myocardial hibernation is an option to minimize the myocardial ischemia and reperfusion injury during cardiac arrest and cardiopulmonary resuscitation. PMID- 17114983 TI - Therapeutic hypothermia after cardiac arrest: unintentional overcooling is common using ice packs and conventional cooling blankets. AB - OBJECTIVES: Although therapeutic hypothermia for cardiac arrest survivors has been shown to improve neurologically intact survival, optimal methods to ensure controlled induction and maintenance of cooling are not clearly established. Precise temperature control is important to evaluate because unintentional overcooling below the consensus target range of 32-34 degrees C may place the patient at risk for serious complications. We sought to measure the prevalence of overcooling (<32 degrees C) in postarrest survivors receiving primarily noninvasive cooling. DESIGN: Retrospective chart review of postarrest patients. SETTING: Three large teaching hospitals. PATIENTS: Cardiac arrest survivors receiving therapeutic hypothermia. INTERVENTIONS: Charts were reviewed if primarily surface cooling was used with a target temperature goal between 32 degrees C and 34 degrees C. MEASUREMENTS AND MAIN RESULTS: Of the 32 cases reviewed, overcooling lasting for >1 hr was identified as follows: 20 of 32 patients (63%) reached temperatures of <32 degrees C, 9 of 32 (28%) reached temperatures of <31 degrees C, and 4 of 32 (13%) reached temperatures of <30 degrees C. Of those with overcooling of <32 degrees C, 6 of 20 (30%) survived to hospital discharge, whereas of those without overcooling, 7 of 12 (58%) survived to hospital discharge (p = not significant). CONCLUSIONS: The majority of the cases reviewed demonstrated unintentional overcooling below target temperature. Improved mechanisms for temperature control are required to prevent potentially deleterious complications of more profound hypothermia. PMID- 17114984 TI - Intrathoracic pressure regulation for intracranial pressure management in normovolemic and hypovolemic pigs. AB - OBJECTIVE: To evaluate the potential to use subatmospheric intrathoracic pressure to regulate intracranial pressure (ICP) in normovolemic and hypovolemic animals, we tested the hypothesis that mechanical devices designed to reduce intrathoracic pressure will decrease ICP in a dose-related manner. An inspiratory impedance threshold device was used in spontaneously breathing animals and an intrathoracic pressure regulator was attached to a positive pressure ventilator and used in apneic animals: both devices lower intrathoracic pressure. DESIGN: Prospective, randomized animal study. SETTING: Animal laboratory facilities. SUBJECTS: A total of 36 female farm pigs in four different protocols (n = 12, 6, 12, and 6, respectively). INTERVENTIONS, MEASUREMENTS, AND MAIN RESULTS: In all protocols, endotracheal, right atrial, central aortic, and ICP were measured continuously. In protocol 1, spontaneously breathing animals were randomized to breath for 15 mins through an impedance threshold device with a cracking pressure of -10 or -15 mm Hg. In protocol 2, after untreated ventricular fibrillation for 4 mins and successful defibrillation to a normal rhythm, spontaneously breathing pigs were used to evaluate the effect of two different impedance threshold device cracking pressures (-10 and -15 mm Hg) on increased ICP. In protocol 3, the acute effects of an intrathoracic pressure regulator on ICP were evaluated in combination with a positive pressure mechanical ventilator in apneic hypovolemic hypotensive pigs after 35% or 50% blood loss. In protocol 4, after 40% blood loss, an intrathoracic pressure regulator was applied for 120 mins and ICP was recorded to determine whether the intrathoracic pressure regulator effects were sustained over time. Inspiratory impedance successfully decreased ICP in spontaneously breathing pigs in a dose-dependent manner and decreased elevated ICP immediately after cardiac arrest and successful resuscitation. The same effect was seen in apneic animals with the use of the intrathoracic pressure regulator. The effect was more pronounced in hypovolemia, and it was sustained for >/=2 hrs. CONCLUSIONS: Reduction of intrathoracic pressure to subatmospheric levels resulted in an instantaneous and sustained reduction in ICP in spontaneously breathing and apneic animals. The effect was most pronounced in the hypovolemic animals. PMID- 17114985 TI - [Persistent unsealed sclerotomy after intravitreal injection of triamcinolone acetonide with a 30-gauge needle]. AB - We report the case of a persistent unsealed sclerotomy following intravitreous injection of triamcinolone through the pars plana using a 30-gauge needle. The injection was made for the treatment of diabetic macular edema. The eye had been treated 9 months before by pars plana vitrectomy and pan retinal photocoagulation for vitreous hemorrhage complicating diabetic proliferative retinopathy. Five days after injection, the patient presented with severe hypotony and chemosis. A conjunctival Seidel was noted at the site of injection. Surgical exploration revealed a punctiform scleral wound without vitreous incarceration, which was closed with a 10-0 nylon suture. No other complication occurred. Our observation highlights that in spite of its simplicity, intravitreous injection of triamcinolone is an invasive procedure that requires rigorous follow-up. In vitrectomized eyes, the higher risk of unsealed scleral wound after intravitreous injection should encourage this injection to be made in a site where no sclerotomy usually occurs. The 6'o-clock meridian could be a good location. PMID- 17114986 TI - [Choroidal white dots and sarcoidosis]. AB - INTRODUCTION: Sarcoidosis is a multisystemic disease characterized by a noncaseating granuloma. Posterior segment disease with white choroidal dots occurs in only 5% of sarcoidosis cases with ocular involvement. OBSERVATION: We report here the case of a 45-year-old Caucasian patient complaining of a skin rash. The biomicroscopic examination of the posterior poles revealed bilateral choroidal white dots with kidney insufficiency. A kidney and skin biopsy revealed the diagnosis of sarcoidosis. CONCLUSION: This case is a rare example of posterior segment manifestation in sarcoidosis that must be considered when observing white choroidal dots. PMID- 17114987 TI - [Meningococcus endophthalmitis without meningitidis]. AB - Meningococcus endophthalmitis is exceptional. We report a case of ocular damage following type C meningococcus septicemia with no meningitis. A 20-year-old man reported to the emergency unit for polyarthritis pain in various joints, associated with chills, nausea, and diarrhea without fever. Ophthalmological examination revealed uveitis. A few days later, endogenous endophthalmitis was suggested because of a worsening general condition and fever spells to 39 degrees C. A hemoculture sampled on the patient's admission 4 days earlier revealed Neisseria meningitidis positivity. Meningococcus septicemia with no meningitis was diagnosed. Before the introduction of antibiotics, meningococcus meningitis was unfortunately frequent and ocular septic embolism was not a rare occurrence. The diagnosis of meningococcemia was delayed in our patient because of the atypical symptomatology and ocular manifestations in the forefront. As with any endogenous endophthalmitis, prognosis is bleak and it should be raised whenever suspected uveitis does not react to standard treatment. PMID- 17114988 TI - [Leukemic infiltration of the optic nerve]. AB - INTRODUCTION: We report a case of unilateral leukemic infiltration of the optic nerve caused by acute lymphoblastic leukemia. CASE REPORT: A 30-year-old woman with acute lymphoblastic leukemia beginning 3 months before was referred for left visual loss. The left fundus showed optic disc engorgement by infiltrating tumor, with numerous hemorrhages and exudates caused by optic nerve leukemic infiltration. CONCLUSION: Optic nerve leukemic infiltration is considered as central nervous system damage with severe prognosis. Treatment is based upon systemic and intrathecal chemotherapy and central nervous system radiation. PMID- 17114989 TI - [Iridoschisis and angle-closure glaucoma: a case report]. AB - INTRODUCTION: Iridoschisis is a rare condition where a localized area of iris stroma is cleaved in two, with the anterior atrophic portion disintegrating into fibrils. It is frequently associated with angle closure glaucoma and with cataract. CASE REPORT: A 70-year-old male presented with reduction of visual acuity in the left eye. Visual acuity was 20/25 OD and 20/70 OS. He had suffered acute angle-closure glaucoma in the right eye 10 years before, treated with laser iridotomy and topical antiglaucomatous therapy. Intraocular pressure was 12 mmHg OD and 46 mmHg OS. Slit lamp biomicroscopic examination revealed bilateral shallow anterior chamber, iris atrophy, and leaf degeneration of the iris stroma. A-scan and B-scan standardized echography in both eyes showed shortening of the anterior chamber, a thickened and more anteriorly located lens, the presence of a few hyperechogeneous flocculated masses floating in the vitreous, and optic disc excavation. CONCLUSION: The mechanism by which iridoschisis causes angle closure is unclear. A pupillary block could be favored by the thickening and anteriorization of the lens and the ciliary processes. Both the presence of strings of iris stroma occluding the trabecula and pupillary block could cause an elevation in intraocular pressure. PMID- 17114990 TI - [Comparison of macular limited translocation and photodynamic therapy for management of choroidal neovascularization in degenerative myopia: a retrospective study]. AB - PURPOSE: To analyze the results of limited macular translocation (MT) and photodynamic therapy (PDT) in subfoveal choroidal neovascularization due to degenerative myopia, with a minimum follow-up of 12 months. METHODS: Retrospective review of 55 consecutive patients: 31 eyes were treated using PDT and 24 were operated on with the limited macular translocation technique with chorioscleral infolding described by de Juan. Before and after each treatment, a complete examination comprised visual acuity, fundus examination, fluorescein angiography, and optical coherence tomography. Mean follow-up was 14 months in the PDT group and 19 months in the MT group. RESULTS: The improvement in visual acuity was better in the MT group than in the PDT group and was correlated with younger age (p<0.001). At month 12, visual acuity improved by 3 ETDRS lines or more in 6 six eyes of the PDT group (19%), with no improvement over 6 lines. In the MT group, visual acuity improved by 3 lines in 14 eyes (58%) including 8 eight eyes (33%) with an improvement of 6 lines or more. Final visual acuity remain unchanged (+/-2 lines) in 16 eyes of the PDT group (66%) and 8 eight eyes of the MT group (33%), and decreased in 9 nine eyes in the PDT group (29%) versus 2 two eyes in the MT group (8%). Mean gain in visual acuity at 12 months was +3.5 ETDRS lines in the MT group and -0.1 line in the PDT group (p=0.001). The mean displacement of the fovea after translocation was 950 microm. The mMean number of PDT treatments was 2.3 during the 12-month follow-up. CONCLUSIONS: Limited macular translocation allowed provided a significant improvement in visual acuity in some eyes with subfoveal neovascularization in myopia, especially in young patients, and resulted in a moderate rate of complications. Longer follow-up and further controlled and randomized studies are required to confirm these encouraging findings. PMID- 17114991 TI - [Late recurrent retinal detachment after scleral buckling]. AB - PURPOSE: To analyze the physiopathologic mechanism, therapeutic modalities, and prognosis of late recurrent retinal detachment. METHODS: A retrospective study was conducted on late recurrent retinal detachment operated with episcleral surgery over a 15-year period. Ten patients were included in this study. RESULTS: Late recurrent retinal detachments occurred in 0.39% of all retinal detachments repaired by episcleral surgery over 15 years. Redetachment occurred 3-7 years after surgery, with etiologies including new retinal breaks (seven cases), reopening of old breaks (three cases), and removal of scleral explant (one case). Proliferative vitreoretinopathy (PVR) grade B was seen in three cases, grade C in six cases, and grade D in one case. After reoperation, the retina was reattached in nine cases. Three eyes were treated with scleral buckle, the others underwent vitreoretinal surgery. Failure occurred in one patient who had a very high level of vitreoretinal proliferation. CONCLUSION: Late recurrent retinal detachments are rare and vitreous base traction seems to be an important factor, although the associated PVR was probably a secondary factor. Treatment depends on PVR, with vitreoretinal surgery necessary in some cases. They usually have a good prognosis. PMID- 17114992 TI - [Macular fold following retinal detachment surgery]. AB - INTRODUCTION: The occurrence of a retinal fold after vitrectomy for retinal reattachment is not often described. In this case report, its treatment, prevention, and mechanism are discussed. CASE REPORT: A 45-year-old pseudophakic woman experienced a superotemporal retinal detachment with macula-on and numerous retinal tears. The treatment consisted of a vitrectomy, perfluorocarbon injection, cryotherapy, and a complete fluid/gas exchange. Postoperative examination disclosed a retinal fold centered by the fovea. The treatment of this macular fold included the creation of a new detachment of the posterior pole by means of an injection of balanced saline solution into the subretinal space through a 39-gauge cannula, the injection of perfluorocarbon liquid to move the fold superiorly, endolaser at the inferior limit of the fold, and silicone oil injection. At the postoperative examination, the macula was flattened and the fold was near the superior temporal arcade. DISCUSSION: The formation of a macular fold after vitrectomy was probably caused by the complete fluid/gas exchange, which displaced the subretinal fluid from the periphery to the posterior pole, detaching the macula. Tangential traction exerted by the presence of intravitreal gas and subretinal fluid might have stretched the retina and resulted in the formation of the fold. This mechanism is similar to the technique used in macular translocation surgery. To avoid this complication in macula-on retinal detachment, we suggest not using perfluorocarbon liquid systematically and replacing the complete fluid/gas exchange with a limited bubble of expansive gas combined with postoperative positioning. PMID- 17114993 TI - [Keratoconus: correlations between clinical aspects and Orbscan evaluations]. AB - PURPOSE: To look for potential correlations between clinical and topographical aspects of keratoconus evaluated by Orbscan (Chauvin-Bausch & Lomb). METHODS: We analyzed 228 keratoconic eyes in a retrospective study over a period of 16 months. The best corrected visual acuity (BCVA) and the corneal opacities were correlated with 26 topographical parameters evaluated by Orbscan. RESULTS: The maximal keratometry is the parameter that correlated best with clinical aspects. The other well-correlated keratometric parameters were the SimKmax and the Mean Power in the 3-mm zone. The distance between the central point and the point with maximal keratometry is the best localization parameter. The maximal amplitude of elevation on the anterior elevation map and the value of the minimal pachymetry also correlate well with the BCVA. The results were nevertheless quite scattered, but could be explained by bias, which will be corrected in further prospective studies. CONCLUSION: Orbscan provides useful information to assess the topography of keratoconus. Some parameters correlate well with clinical aspects, which may lead to a new clinico-topographical classification. More investigations are necessary to reduce the scattering of the results. PMID- 17114994 TI - [Photodynamic therapy of circumscribed choroidal hemangiomas]. AB - PURPOSE: To evaluate the safety and efficacy of photodynamic therapy in the management of symptomatic circumscribed choroidal hemangiomas (CCH) with exudative activity. PATIENTS AND METHODS: This prospective nonrandomized study included eight patients referred to us for management of CCH with exudative activity. PDT was applied by diode laser (689 nm) with a light dose of 100 J/cm2 and using verteporfin (6 mg/m2) IV bolus injection. The treatment spot diameter accurately corresponded to the tumor basal diameter (TBD). RESULTS: The mean tumor thickness before PDT was 3.3 mm (range, 2.7-7.5 mm) and the mean largest TBD was 6.1 mm (range, 5-12 mm). All patients were managed with a single PDT application. After a mean follow-up of 24.2 months (range, 17-29 months), all CCH (100%) showed regression of tumor thickness and complete resorption of the serous retinal fluid; 87.5% of the patients showed a flat tumor scar. All CCH showed overlying PDT-related retinal pigmented epithelium atrophic changes. No case of retinal vascular occlusion or recurrent leakage was documented. Five patients (62.5%) showed visual improvement, one (12.5%) retained stable vision and two (25%) developed worse vision due to chronic exudation-related RPE changes in one patient and preretinal fibrosis in another. CONCLUSION: PDT using verteporfin offers a safe and effective therapeutic option to manage CCH. Complete resorption of subretinal fluid is usually associated with visual improvement. PMID- 17114995 TI - [Ocular manifestations associated with nephronophthisis and genetic study in three Tunisian families]. AB - PURPOSE: Nephronophthisis is a familial interstitial nephropathy with an autosome recessive mode of transmission. In some cases, it is associated with ocular manifestations such as retinitis pigmentosa in Senior-Loken syndrome. We report ocular abnormalities and genetic results in three affected Tunisian families. PATIENTS AND METHODS: Twenty-two members of these three families underwent a complete ophthalmologic examination (visual acuity, slit lamp biomicroscopy, ophthalmoscopy, and retinal electrophysiology). For genetic study, all individuals were genotyped and underwent a genomic sequence. RESULTS: Twenty-two subjects, nine of whom presented nephronophthisis, were included in this study. Retinitis pigmentosa was found in three cases. Our genetic study demonstrated that patients belonging to family 1 had homozygous deletions in NPHP1, all affected individuals from family 3 were linked to NPHP4 and presented a deletion in exons 2 and 3. Results are pending for patients in family 2. CONCLUSION: Senior-Loken syndrome is a rare hereditary disease that combines familial juvenile nephronophthisis and retinitis pigmentosa. This association was described in the literature in 39%-43% of cases. In our study, it was approximately 33% of cases. The genetic study can sometimes obviate the need for renal puncture, especially when the homozygous deletion of NPHP1 gene is confirmed. PMID- 17114996 TI - [Experience with 25-gauge transconjunctival vitrectomy compared to a 20-gauge system. Analysis of 132 cases]. AB - PURPOSE: The transconjunctival sutureless vitrectomy (TSV 25) is a new technique showing substantial progress. METHODS: A prospective study of 66 eyes of 66 patients who underwent surgical procedures using the transconjunctival standard vitrectomy system (TSV 25), between June 2004 and June 2005. The results were compared with the results in an equivalent population (66 patients) that underwent vitrectomy with a standard 20-gauge system. RESULTS: The most important differences between groups were 1) shorter surgical time in the 25-gauge vitrectomy group than in the 20-gauge group, with a mean time of 28.15+/-5.50 min for group 1 and 44.15+/-5.64 min for group 2 (p<0.001) and 2) lower mean postoperative intraocular tension in the first group of patients: in this group, the intraocular tension before surgery was a mean of 16.27+/-1.76 mmHg, decreasing to 12.17+/-1.36 mmHg, with a posterior increase to normal values of 15.06+/-1.66 mmHg at 7 days after surgery and a mean of 16.10+/-1.51 mmHg at 15 days. This decrease was not observed in the group of patients who underwent 20 gauge vitrectomy surgery. The other postoperative complications were similar in both groups except for four cases of anterior proliferation and two cases of fibrinous uveitis in group 2. CONCLUSION: The transconjunctival sutureless system (TSV 25) decreases surgical time and postoperative inflammation, with optimal postoperative patient comfort, but currently the accurate selection of patients remains important. PMID- 17114997 TI - [Diagnosis of Acanthamoeba spp. keratitis with PCR]. AB - AIM: Evaluation of a PCR assay as a diagnostic tool for detection of Acanthamoeba spp. in patients presenting infectious keratitis. METHODS: Between August 2001 and November 2002, 342 clinical specimens consisting in corneal scrapings from 334 patients were tested for Acanthamoeba using direct microscopy, culture, and PCR. A fragment of Acanthamoeba 18S rRNA gene was amplified using a set of primers referred to as Nelson's primers. RESULTS: A diagnosis of Acanthamoeba keratitis was considered for nine patients. Amoeba growth in culture was unfruitful for all of these cases. Eight patients had corneal scrapings that tested positive with PCR; in two cases direct microscopy observations confirmed PCR results. For one patient, a negative PCR result was obtained; however, a second corneal sample and cysts staining on May-Grunwald-Giemsa were positive. A false-positive PCR result was noted related to another amebic genus. A risk factor was found in all Acanthamoeba keratitis cases (contact lenses, trauma). Topical treatment was successful, and keratoplasty was necessary afterwards for optical rehabilitation in five patients. CONCLUSION: This study suggests that PCR is a sensitive diagnostic tool, superior to conventional techniques for detecting Acanthamoeba in corneal lesions. PMID- 17114998 TI - [Dosage and kinetics of MMC release of a collagen implant used as a delivery device in glaucoma surgery in the rabbit eye]. AB - PURPOSE: To determine the absorption and the release of mitomycin-C from a collagen implant and tissue impregnation in the anterior structures of the rabbit eye. METHODS: Determining the quantity of mitomycin-C that a collagen implant can absorb with the difference between dry and soaked weight. Mitomycin-C release was measured in vitro using spectrophotometry. The measurement was repeated using a bioassay. Ocular tissue impregnation was determined in 12 eyes of six rabbits. Sclera, implant, aqueous, and ciliary body specimens were collected for concentration measurement using HPLC from 1 to 6 h after surgery. RESULTS: The mean mitomycin-C quantity absorbed in the implant was 3.22+/-0.2 microg. In vitro release was 0.13 mg/ml after 10 min and 0.05 microg/ml at 6 h. The bioassay showed almost no antifibrotic activity in sclera. In vivo release of mitomycin-C was high from the first to the sixth hour. CONCLUSION: After filtering surgery, mitomycin-C in the collagen implant is clearly released and ocular tissues are effectively impregnated. PMID- 17115000 TI - [Lacrimal film and the ocular surface]. AB - Abnormalities of the ocular surface affect a large proportion of the general population. Symptoms depend on whether the lids, the corneoconjunctival surface, or the lacrimal layer are involved. The latter results from the balance of a fragile mixture of water, lipids, and mucus. Different circumstances can lead to the disruption of the lacrimal layer associated with dry-eye symptoms, eventually leading to inflammation. Alterations observed during dry-eye disease, meibomian dysfunction, allergic conjunctivitis, contact lens wear infectious conjunctivitis, or after refractive surgery are reviewed. Patient relief depends on a reliable analysis of the mechanism involved in the ocular surface disease. PMID- 17115001 TI - [Corneal epithelial diseases related to limbal stem cell deficiency]. AB - Treatment of corneal epithelial diseases induced by limbal stem cell deficiency is an important challenge in ocular surface reconstruction. Since the 1990s, corneal stem cells have been localized in the limbus. This new concept completely changed the way we consider ocular surface reconstruction, with new diseases now found to be isolated in the ocular surface. Limbus insufficiency syndromes are specific depending on their origin (congenital or acquired), their expression (unilateral or bilateral, partial or total), their progression (acute or chronic), and the mechanism involved (burn, infection, chronic inflammation, etc.). Some of these diseases are local diseases and others are systemic diseases. Clinically, limbus insufficiency is a switch of the normal corneal epithelial phenotype (expression of a specific keratin, avascularity, and transparency of the corneal matrix) in an opaque and fibrovascularized cornea. In terms of cellular biology, a phenotype is a terminal expression of a cell differentiation process. This process is the outcome of the interaction between the genome of a cell or a group of cells with their microenvironment. In limbus insufficiency, epithelial cells and corneal matrix are destroyed, and it is the destruction of these two components that leads to limbus insufficiency syndrome. PMID- 17115002 TI - [Advantages of amniotic membrane transplantation in eye surface diseases]. AB - Amniotic membrane transplantation is now a widely adopted technique in the field of eye surface diseases. Depending on the indication, the amniotic membrane can be used as either a graft or a patch. When used as a graft, the amniotic membrane serves as a substrate for regrowth of deficient epithelium; the aim is to integrate this membrane. The basal membrane reinforces the adhesion and differentiation of the corneal epithelial cells, facilitates their migration, and prevents their apoptosis. When used as a patch, the amniotic membrane is sutured epithelium-down so as to maximize the concentration of biological factors delivered by this membrane: the membrane covers the diseased cornea and acts as a biological bandage and analgesic. The best indications for amniotic membrane grafts are acute chemical burns and trophic corneal ulcers refractory to all medical treatment. When these ulcers are perforated or in the early stages of perforation, it is best to use multiple layers of amniotic membrane, restoring the thickness of the cornea. In cases of confirmed limbal deficiency, amniotic membrane grafts may be a useful complement to the necessary limbal stem cell grafts. In the future, amniotic membranes will provide an indispensable support for the expansion of cultured stem cells. Amniotic membrane grafts may also be used to reconstruct the conjunctiva following the exeresis of symblepharons or conjunctival tumors. However, the use of this technique is currently limited to diseases with little inflammation and no extensive fibrosis. PMID- 17115005 TI - [Do dendritic cells modified with retroviral vectors carrying cytokine genes become a therapeutic tool?]. AB - For many years, the application of various types of immunostimulators (among which cytokines have proved to play a crucial role) has been one of the ways to enhance anti-tumor immunity. Dendritic cells genetically modified towards cytokine production have been proposed as a potential therapeutic tool to enable restoration of the cytokine environment diminished by a growing tumor. Retroviral vectors have been the most often used carriers of cytokine genes. Therefore, optimization of the genetic procedure of inserting retroviral vectors into DCs has been widely studied largely in order to achieve better immunotherapeutic effects against tumor. PMID- 17115006 TI - The screening analysis of antiradical activity of some plant extracts. AB - INTRODUCTION: There is a need for screening studies in order to select plant extracts or plant raw materials with strong antiradical activity which could be used as medicines or substances to protect food from oxidation. In this paper the antiradical activities of some plant raw materials were investigated. MATERIAL/METHODS: The intensity of antiradical activity of extracts was investigated using DPPH* (1,1-diphenyl-2-picrylhydrazyl) radical as a substrate. The antiradical activity unit was defined and the number of antiradical activity units EAU(515) per 1 mg of plant extract and TAU(515) per 1 g of plant raw materials were calculated. Plant extracts were obtained with a methanol or methanol-water (1:1) solution. RESULTS AND DISCUSSION: The highest numbers of antiradical activity units EAU(515) were found for ethyl acetate extracts from the leaves of green and black tea. The lowest EAU(515) value was demonstrated for garlic extracts. When the number of activity units TAU(515) was calculated per 1 g of raw material, the highest value was found for the leaves of green tea, much lower for bee propolis and the leaves of black tea. On the basis of the presented results, green tea leaves, bee propolis, and the leaves of black tea could be considered as potential sources of extracts with strong antiradical activity. PMID- 17115007 TI - [Mechanisms involved in the regulation of immune response in experimental autoimmune encephalomyelitis in mice]. AB - Multiple Sclerosis (MS) is a chronic inflammatory autoimmune disease of the central nervous system (CNS) characterized by the presence of cellular infiltrates consisting primarily of lymphocytes and macrophages and localized areas of demyelination in the CNS. MS is thought to be initiated by self-reactive CD4(+) Th1 T cells. Thus far, treatment modalities for MS are limited, with the most common acting nonspecifically on the immune system, resulting in a general immunosuppression accompanied by severe side effects. There is a large demand for developing MS therapy that particularly targets pathogenic myelin-specific T cells. Experimental allergic encephalomyelitis (EAE) is a well-characterized animal model that mimics many of the disease symptoms of MS, including the presence of cellular infiltrates and demyelination. EAE can be actively induced in genetically susceptible strains of mice, rats, and monkeys and is mediated by activated autoreactive CD4(+) T cells that are specific to MBP (myelin basic protein). The knowledge acquired using EAE allows us to better understand the pathogenesis of MS and thus manipulate particular components of the immune response in order to develop an efficient therapy. PMID- 17115008 TI - [Dexrazoxane (ICRF-187)--a cardioprotectant and modulator of action of some anticancer drugs]. AB - The nthracycline antibiotics are among the most widely used and effective anticancer drugs. The therapeutic efficacy of this class of drugs is limited by cumulative cardiac toxicity. Dexrazoxane is the only clinically approved cardioprotective agent used in anthracycline-containing anticancer therapy. Its cardioprotective action allows the use of a much higher cumulative dose of anthracyclines and improvement in the effectiveness of treatment. Anthracyclines form complexes with iron ions, which are very active in the production of reactive oxygen species responsible for the lipid peroxidation of mitochondrial and endoplasmatic reticulum membranes. This process seems to be the major cause of anthracycline-induced cardiotoxicity. Dexrazoxane exerts its protective effects by rapid and complete binding of ferric and ferrous ions, even by displacing the metal ions from complexes with anthracyclines. Besides its cardioprotective effect, dexrazoxane also exhibits anticancer properties. Like other derivatives of bisdioxopiperazine, dexrazoxane is a catalytic inhibitor of eukaryotic DNA topoisomerase II, the key enzyme controlling DNA topology and contributing to the replication and transcription processes. Dexrazoxane is able to lock topoisomerase II at the stage of the enzyme reaction cycle where the enzyme forms a closed clamp around the DNA. This phenomenon seems to be the main reason for the generation of DNA double-strand breaks by dexrazoxane as well as its cytotoxicity against quickly proliferating cancer cells. Other effects of its topoisomerase II catalytic inhibition is the induction of cell differentiation and apoptosis. Dexrazoxane may be used not only as a cardioprotective agent, but also as a modulator of action of some anticancer drugs by enhancing their selectivity or by delaying the development of multidrug resistance. PMID- 17115009 TI - [Structure and function of midkine, a novel heparin-binding growth factor]. AB - Midkine is a multifunctional peptide which, together with pleiotrophin, forms a structurally distinct family of heparin-binding growth factors. The paper presents the structure of midkine together with its amino-acid sequence and the functions of its domains as well as structural differences between the physiological forms of this peptide and those found in tumors. The localization of the midkine gene and its tissue expression both in embryonic life and in mature organisms is described. The available information on midkine receptors is discussed in detail. Most often they seem to be proteoglycans containing heparan sulfate and chondroitin sulfate, which can also be a part of the multimolecular receptor complexes binding midkine. The variety of midkine receptors is probably responsible for the diverse biological activity of this peptide. The paper presents up-to-date views on the biological activity of midkine both at the cellular (mitogenic properties, participation in apoptosis, and cellular migration, adhesion, morphological differentiation, and chemokine synthesis stimulation) and tissue levels (participation in organogenesis, tissue regeneration and protection and in the formation and degradation of extracellular matrix). PMID- 17115010 TI - Anaesthetic management in asthma. AB - Anaesthetic management in asthmatic patients has been focused on avoiding bronchoconstriction and inducing bronchodilation. However, the definition of asthma has changed over the past decade. Asthma has been defined as a clinical syndrome characterized by an inflammatory process that extends beyond the central airways to the distal airways and lung parenchyma. With this concept in mind, and knowing that asthma is a common disorder with increasing prevalence rates and severity worldwide, a rational choice of anaesthetic agents and procedures is mandatory. Thus, we pursued an update on the pharmacologic and technical anaesthetic approach for the asthmatic patient. When feasible, regional anaesthesia should be preferred because it reduces airway irritation and postoperative complications. If general anaesthesia is unavoidable, a laryngeal mask airway is safer than endotracheal intubation. Lidocaine inhalation, alone or combined with albuterol, minimizes histamine-induced bronchoconstriction. Propofol and ketamine inhibit bronchoconstriction, decreasing the risk of bronchospasm during anaesthesia induction. Propofol yields central airway dilation and is more reliable than etomidate or thiopental. Halothane, enflurane, and isoflurane are potent bronchodilators and can be helpful even in status asthmaticus. Sevoflurane has shown controversial results in asthmatic patients. Vecuronium, rocuronium, cisatracurium, and pancuronium do not induce bronchospasm, while atracurium and mivacurium can dose-dependently release histamine and should be cautiously administered in those patients. Further knowledge about the sites of action of anaesthetic agents in the lung, allied with our understanding of asthma pathophysiology, will establish the best anaesthetic approach for people with asthma. PMID- 17115012 TI - Open intensive care units: the case in favour. AB - Intensive care units traditionally have a closed structure in Italy. They generally have highly restrictive visiting policies, limiting the admission and attendance of family members. This article deals with the issue of open intensive care unit (ICU), i.e. a unit oriented towards the implementation of non restrictive visiting policies and committed to removing all barriers that have no justifiable necessity, on the level of time, on the physical level and on the level of relationships. The most common objections to opening intensive care units are examined, and the clinical and ethical reasons behind this alternative are considered. As things stand, there is no solid scientific basis for limiting visitors' access to ICUs and keeping ICUs ''closed''. On the contrary, opening ICUs offers a strategy which is to patients' advantage. Opening ICUs should come about not so much in answer to pressure generated by a growing social awareness, or in simple recognition of a right, but because this policy addresses more comprehensively the issue of respect for the patient, as well as providing more appropriate responses to many needs of both patients and families. It is a decision which requires doctors and nurses to rethink their relationships with patients and their families, which calls for original solutions for each individual situation, and which should be subject to periodic checks. PMID- 17115013 TI - BIS - AAI and clinical measures during propofol target controlled infusion with Schnider's pharmacokinetic model. AB - AIM: The A-line autoregressive index (AAI) and the Bispectral Index Score (BIS) are two commercially available indexes of anesthetic depth widely used in clinical practice. The aim of the current study was to compare the accuracy of AAI, BIS, Schnider's predicted effect-site concentration of propofol (Ce propofol) to assess depth of anesthesia. METHODS: Forty-four patients scheduled for major elective abdominal surgery received target effect-site controlled infusion of propofol. Target effect-site (Ce propofol) was started at 1.5 mug/mL and increased every 4 min by 1.0 microg/mL until 5.5 microg/mL were achieved. At every step sedation level was estimated, using AAI, BIS, Observer's Assessment of Alertness/Sedation scale (OAA/S), loss of eyelash reflex and Ce propofol. RESULTS: We enrolled 44 patients, 20 males and 24 females, ASA I/II 18/26, 48+/ 10 years, 68.2+/-9 kg, 165+/-7.1 cm, body mass index (BMI) 25+/-3.5. At increasing Ce propofol BIS-AAI values decreased progressively (BIS range 97-38) (AAI range 97-17). Values of BIS < or = 50, of AAI < or = 48 and of Ce propofol > or = 5.1 resulted in OAA/S=0, while values of BIS < or = 62, AAI < or = 53 and Ce propofol < or = 3.5 resulted in OAA/S=2. Loss of eyelash reflex occurred when values were BIS < or = 64 and AAI < or 61. CONCLUSION: BIS, AAI, propofol site effect concentration revealed information on sedation level and consciousness but no gold standard yet exists because of consistent overlap between ''conscious'' and ''not conscious'' states. PMID- 17115014 TI - Severe scombroid fish poisoning syndrome requiring aggressive fluid resuscitation in the emergency department: two case reports. AB - Scombroid fish poisoning (scombrotoxism, scombroid ichthyotoxicosis) is a food related illness typically associated with the consumption of dark and white meat fish. Two patients presented to the emergency department. Metilprednisone 1000 mg and ranitidine 150 mg were administered initially. A large amount of crystalloids and colloids in in combination with vasoactive drugs were required to maintain normopressure. Levels of histamine and N-methylhistamine were far above the normal mean. Carboxyhemoglobin levels were also tested to exclude a superimposition of carbon monoxide intoxication. In both cases, major symptoms occurred and were treated aggressively. Early goal directed fluid therapy corrected the DO2/VO2 unbalance, due to a distributive pattern of hypovolemic impending shock, and permitted a rapid stabilisation of both patients. It is important to recognize the syndrome as an intoxication (rather than an allergic reaction) so that the source of the toxin can be identified and further cases prevented. It is also important to investigate where the fish was cooked (i.e. in an open space vs. closed space), to exclude the possibility of a concomitant carbon monoxide intoxication, which would require transfer the patient to a hospital facility equipped with a hyperbaric chamber. PMID- 17115015 TI - Parenteral nutrition in ventilated patients with chronic obstructive pulmonary disease: long chain vs medium chain triglycerides. AB - AIM: The aim of this study was to assess the usefulness of a lipid formulation containing a physical mixture of medium (MCT) and long chain triglycerides (LCT) compared with a long chain triglycerides emulsion in patients affected by chronic obstructive pulmonary disease with acute respiratory failure. METHODS: Twenty four patients requiring mechanical ventilation were randomly selected in 2 groups and received total parenteral nutrition. Twelve patients received a MCT/LCT emulsion (50:50), the others used a 100% LCT emulsion. Nutritional status, metabolic rate, time of ventilatory support and weaning were evaluated. RESULTS: Both groups showed an improvement of all nutritional parameters evaluated; oxygen uptake, carbon dioxide output and respiratory gas exchange ratio were similar in both groups. The duration of mechanical ventilation was not significantly different; however, the time of weaning in the MCT/LCT group was significantly shorter. The longer weaning time in the LCT group patients could be related to vasoactive intermediates deriving from long chain fatty acids. The T-cell subsets, which were evaluated for both groups, showed a significant decrease of T helper-T suppressor ratio in the LCT group. CONCLUSION: MCT/LCT emulsion is an effective lipid supplementation and should be considered the therapy of choice in COPD patients; however, the relationship between lipid emulsions administered and length of weaning requires further investigations. PMID- 17115016 TI - Nurses' knowledge and application of evidence-based guidelines for preventing ventilator-associated pneumonia. AB - AIM: The aim of the study was to evaluate the nurses' knowledge and to highlight the causes that hinder guidelines implementation. METHODS: EXPERIMENTAL DESIGN: descriptive study. SETTING AND PARTICIPANTS: 106 nurses working in the ICUs of a major Italian hospital of national importance. INTERVENTION: administration of a questionnaire listing 21 non-pharmacological strategies considered the most useful in the literature. RESULTS: Eighty-four nurses responded to the questionnaire. Only 19 (22.6%) declared that their knowledge of ventilation associated pneumonia (VAP) and the strategies used to prevent it were satisfactory, whereas 46 (54.8%) declared that they were poorly informed; 68 nurses (80.9%) said that they applied one or more strategies, and 15 (17.9%) that they applied none. The reasons given for not applying the strategies were: method not foreseen in Department protocols (31.5%), lack of the necessary resources (14.3%), disagreement with the method (3.2%), high costs (2.6%), the possibility of causing discomfort (1%) or side effects (0.6%). CONCLUSIONS: In our experience, VAP preventive strategies are widely applied by nurses, but not in a responsible and informed manner. It is important to ensure that nurses receive continuous training and are involved in drawing up and updating Departmental protocols and guidelines for care and behaviour. PMID- 17115017 TI - Metastatic renal cell carcinoma presenting as a chalazion. PMID- 17115018 TI - One-stage correction for blepharophimosis syndrome. AB - PURPOSE: To classify the severity of blepharophimosis, describe associated features and their effects on the incidence of amblyopia and to recommend guidelines for surgical treatment and management of surgical complications. METHODS: The case records of 23 patients with blepharophimosis syndrome were examined retrospectively. Patients' photographs and measurements were reviewed to analyse the severity of blepharophimosis, surgical techniques undertaken, surgical outcomes, and complications. Statistical analyses were performed using paired-sample t-tests to evaluate the surgical outcome and Spearman correlation to examine the influence of blepharophimosis on the interpalpebral fissure height (PFH). RESULTS: Eighteen out of 23 (78%) patients underwent one-stage surgery before the age of 5 years. About 31% of these patients had amblyopia. Only two patients had a blepharophimosis ratio greater than 1.5 as poor result. Two out of 18 (11%) patients with PFHs more than 2 mm needed a repeat operation, but all five (100%) patients with s less than 2 mm (very severe ptosis) needed repeat operations. CONCLUSIONS: The one-stage corrective procedure provided acceptable results both in function and cosmesis. However, patients with very severe ptosis required multiple stages of reconstruction for ptosis correction at an earlier age, after which correction of telecanthus and small horizontal palpebral fissure length followed at an older age. PMID- 17115019 TI - Surgical removal of massive macular hard exudate combined with intravitreal triamcinolone in diabetic maculopathy. PMID- 17115020 TI - First-line therapy with latanoprost 0.005% results in improved ocular circulation in newly diagnosed primary open-angle glaucoma patients: a prospective, 6-month, open-label study. AB - PURPOSE: To evaluate the effect of latanoprost 0.005% on the optic nerve head (ONH) and retinal circulation of newly diagnosed and previously untreated primary open-angle glaucoma (POAG) patients. METHODS: Twenty-two newly diagnosed and previously untreated POAG patients (mean age+/-SD: 68.38+/-11.92 years) were included in this longitudinal open-label study. Patients were treated with latanoprost 0.005% once a day. Intraocular pressure (IOP), systemic blood pressure (BP), mean ocular perfusion pressure (MOPP), and ocular perfusion parameters 'volume', 'velocity', and 'flow' measured at the optic nerve head (ONH) and retina by means of Heidelberg Retina Flowmeter system were evaluated during a 6-month follow-up period. RESULTS: Treatment with latanoprost 0.005% resulted in a significant decrease in IOP (P<0.0001) and increase in MOPP (P<0.0001). After correcting for changes in MOPP, the blood velocity measured at the ONH level was significantly higher after 6 months of treatment than at baseline (P=0.0310). In addition, blood volume and flow measured at the peripapillary retina level improved after 3 and 6 months of treatment (P=0.0170; P=0.0260, and P=0.0170; P=0.0240 respectively). CONCLUSION: Previously untreated POAG patients exhibit reduced IOP, increased MOPP and improved ocular perfusion at the ONH and retina levels when treated with Latanoprost 0.005%. These effects could be beneficial for glaucoma patients suffering from ocular vascular dysregulation. PMID- 17115021 TI - 'Late' functionally successful repair of Descemet's membrane detachment following phacoemulsification. PMID- 17115022 TI - A case of recurrent infectious crystalline keratopathy secondary to Haemophilus influenzae. PMID- 17115023 TI - Expression profiles associated with aggressive behavior in Merkel cell carcinoma. AB - Primary neuroendocrine carcinoma of the skin, or Merkel cell carcinoma, is the most aggressive cutaneous neoplasm. In spite of its similarities to small cell carcinomas from other locations, Merkel cell carcinoma shows many peculiarities probably related to its epidermal origin and the etiologic role of UV radiation. We have immunohistochemically investigated 43 markers on a tissue microarray in which 31 surgically resected Merkel cell carcinomas were represented. Of these, 15 patients remained free of disease after removal, whereas 16 developed metastases. Immunoreactivity was scored according to staining intensity and the percentage of positive cells. We found statistically significant correlations between metastatic tumor spread and overexpression of matrix metalloproteinase (MMP) 7, MMP10/2, tissue inhibitor of metalloproteinase 3, vascular endothelial growth factor (VEGF), P38, stromal NF-kappaB, and synaptophysin. Also detected were statistically significant correlations between the expression levels of MMP7 and VEGF, MMP7 and P21, MMP7 and P38, MMP10/2 and VEGF, P38 and synaptophysin, P38 and P53, and P21 and stromal NF-kappaB. These findings may be helpful in predicting the clinical course of Merkel cell carcinoma and are potentially useful for the development of targeted therapies. PMID- 17115024 TI - The flexibility window in zeolites. AB - Today synthetic zeolites are the most important catalysts in petrochemical refineries because of their high internal surface areas and molecular-sieving properties. There have been considerable efforts to synthesize new zeolites with specific pore geometries, to add to the 167 available at present. Millions of hypothetical structures have been generated on the basis of energy minimization, and there is an ongoing search for criteria capable of predicting new zeolite structures. Here we show, by geometric simulation, that all realizable zeolite framework structures show a flexibility window over a range of densities. We conjecture that this flexibility window is a necessary structural feature that enables zeolite synthesis, and therefore provides a valuable selection criterion when evaluating hypothetical zeolite framework structures as potential synthetic targets. We show that it is a general feature that experimental densities of silica zeolites lie at the low-density edge of this window--as the pores are driven to their maximum volume by Coulomb inflation. This is in contrast to most solids, which have the highest density consistent with the local chemical and geometrical constraints. PMID- 17115025 TI - FeCo/graphitic-shell nanocrystals as advanced magnetic-resonance-imaging and near infrared agents. AB - Nanocrystals with advanced magnetic or optical properties have been actively pursued for potential biological applications, including integrated imaging, diagnosis and therapy. Among various magnetic nanocrystals, FeCo has superior magnetic properties, but it has yet to be explored owing to the problems of easy oxidation and potential toxicity. Previously, FeCo nanocrystals with multilayered graphitic carbon, pyrolytic carbon or inert metals have been obtained, but not in the single-shelled, discrete, chemically functionalized and water-soluble forms desired for biological applications. Here, we present a scalable chemical vapour deposition method to synthesize FeCo/single-graphitic-shell nanocrystals that are soluble and stable in water solutions. We explore the multiple functionalities of these core-shell materials by characterizing the magnetic properties of the FeCo core and near-infrared optical absorbance of the single-layered graphitic shell. The nanocrystals exhibit ultra-high saturation magnetization, r1 and r2 relaxivities and high optical absorbance in the near-infrared region. Mesenchymal stem cells are able to internalize these nanoparticles, showing high negative contrast enhancement in magnetic-resonance imaging (MRI). Preliminary in vivo experiments achieve long-lasting positive-contrast enhancement for vascular MRI in rabbits. These results point to the potential of using these nanocrystals for integrated diagnosis and therapeutic (photothermal-ablation) applications. PMID- 17115026 TI - Lsh is required for meiotic chromosome synapsis and retrotransposon silencing in female germ cells. AB - Lymphoid specific helicase (Lsh) is a major epigenetic regulator that is essential for DNA methylation and transcriptional silencing of parasitic elements in the mammalian genome. However, whether Lsh is involved in the regulation of chromatin-mediated processes during meiosis is not known. Here, we show that Lsh is essential for the completion of meiosis and transcriptional repression of repetitive elements in the female gonad. Oocytes from Lsh knockout mice exhibit demethylation of transposable elements and tandem repeats at pericentric heterochromatin, as well as incomplete chromosome synapsis associated with persistent RAD51 foci and gammaH2AX phosphorylation. Failure to load crossover associated foci results in the generation of non-exchange chromosomes. The severe oocyte loss observed and lack of ovarian follicle formation, together with the patterns of Lsh nuclear compartmentalization in the germ line, demonstrate that Lsh has a critical and previously unidentified role in epigenetic gene silencing and maintenance of genomic stability during female meiosis. PMID- 17115027 TI - Cyclin E-Cdk2 temporally regulates centrosome assembly and establishment of polarity in Caenorhabditis elegans embryos. AB - Establishment of polarity in C. elegans embryos is dependent on the centrosome. The sperm contributes a pair of centrioles to the egg and these centrioles remain incapable of polarizing the cortex while the egg completes meiosis. Coincident with the establishment of polarity, the centrioles recruit centrosomal proteins, several of which are required for polarity, suggesting that the temporal regulation of centrosome assembly may control the initiation of polarization. We found that cyclin E-Cdk2 is required for the establishment of polarity. Cyclin E Cdk2 controls the recruitment of centrosomal proteins specifically at the time of polarity establishment. Cyclin E is required for several examples of asymmetric cell division and fate determination in C. elegans and Drosophila. Here, we suggest a possible mechanism for cyclin E-Cdk2-dependent differentiation: the establishment of cortical polarity by the centrosome. PMID- 17115028 TI - Numb is a suppressor of Hedgehog signalling and targets Gli1 for Itch-dependent ubiquitination. AB - The developmental protein Numb is a major determinant of binary cell fates. It is also required for the differentiation of cerebellar granule cell progenitors (GCPs) at a stage of development responsive to the morphogenic glycoprotein Hedehog. Hedgehog signalling is crucial for the physiological maintenance and self-renewal of neural stem cells and its deregulation is responsible for their progression towards tumorigenesis. The mechanisms that inhibit this pathway during the differentiation stage are poorly understood. Here, we identify Numb as a Hedgehog-pathway inhibitor that is downregulated in early GCPs and GCP-derived cancer cells. We demonstrate that the Hedgehog transcription factor Gli1 is targeted by Numb for Itch-dependent ubiquitination, which suppresses Hedgehog signals, thus arresting growth and promoting cell differentiation. This novel Numb-dependent regulatory loop may limit the extent and duration of Hedgehog signalling during neural-progenitor differentiation, and its subversion may be a relevant event in brain tumorigenesis. PMID- 17115029 TI - A novel function of Drosophila eIF4A as a negative regulator of Dpp/BMP signalling that mediates SMAD degradation. AB - Signalling by the TGF-beta superfamily member and BMP orthologue Decapentaplegic (Dpp) is crucial for multiple developmental programmes and has to be tightly regulated. Here, we demonstrate that the Drosophila Dpp pathway is negatively regulated by eukaryotic translation initiation factor 4A (eIF4A), which mediates activation-dependent degradation of the Dpp signalling components Mad and Medea. eIF4A mutants exhibit increased Dpp signalling and accumulation of Mad and phospho-Mad. Overexpression of eIF4A decreases Dpp signalling and causes loss of Mad and phospho-Mad. Furthermore, eIF4A physically associates with Mad and Medea, and promotes their degradation following activation of Dpp signalling in a translation-independent manner. Finally, we show that eIF4A acts synergistically with, but independently of, the ubiquitin ligase DSmurf, indicating that a dual system controls SMAD degradation. Thus, in addition to being an obligatory component of the cap-dependent translation initiation complex, eIF4A has a novel function as a specific inhibitor of Dpp signalling that mediates the degradation of SMAD homologues. PMID- 17115030 TI - The armadillo protein p0071 regulates Rho signalling during cytokinesis. AB - Cytokinesis requires the spatio-temporal coordination of cell-cycle control and cytoskeletal reorganization. Members of the Rho-family of GTPases are crucial regulators of this process and assembly of the contractile ring depends on local activation of Rho signalling. Here, we show that the armadillo protein p0071, unlike its relative p120(ctn), is localized at the midbody during cytokinesis and is essential for cell division. Both knockdown and overexpression of p0071 interfered with normal cell growth and survival due to cytokinesis defects with formation of multinucleated cells and induction of apoptosis. This failure of cytokinesis seemingly correlated with the deregulation of Rho activity in response to altered p0071 expression. The function of p0071 in regulating Rho activity occurred through an association of p0071 with RhoA, as well as the physical and functional interaction of p0071 with Ect2, the one Rho guanine nucleotide exchange factor (GEF) essential for cytokinesis. These findings support an essential role for p0071 in spatially regulating restricted Rho signalling during cytokinesis. PMID- 17115031 TI - Regulation of cell shape by Cdc42 is mediated by the synergic actin-bundling activity of the Eps8-IRSp53 complex. AB - Actin-crosslinking proteins organize actin into highly dynamic and architecturally diverse subcellular scaffolds that orchestrate a variety of mechanical processes, including lamellipodial and filopodial protrusions in motile cells. How signalling pathways control and coordinate the activity of these crosslinkers is poorly defined. IRSp53, a multi-domain protein that can associate with the Rho-GTPases Rac and Cdc42, participates in these processes mainly through its amino-terminal IMD (IRSp53 and MIM domain). The isolated IMD has actin-bundling activity in vitro and is sufficient to induce filopodia in vivo. However, the manner of regulation of this activity in the full-length protein remains largely unknown. Eps8 is involved in actin dynamics through its actin barbed-ends capping activity and its ability to modulate Rac activity. Moreover, Eps8 binds to IRSp53. Here, we describe a novel actin crosslinking activity of Eps8. Additionally, Eps8 activates and synergizes with IRSp53 in mediating actin bundling in vitro, enhancing IRSp53-dependent membrane extensions in vivo. Cdc42 binds to and controls the cellular distribution of the IRSp53-Eps8 complex, supporting the existence of a Cdc42-IRSp53-Eps8 signalling pathway. Consistently, Cdc42-induced filopodia are inhibited following individual removal of either IRSp53 or Eps8. Collectively, these results support a model whereby the synergic bundling activity of the IRSp53-Eps8 complex, regulated by Cdc42, contributes to the generation of actin bundles, thus promoting filopodial protrusions. PMID- 17115032 TI - Tyrosine phosphorylation controls PCNA function through protein stability. AB - The proliferating cell nuclear antigen (PCNA) is an essential protein for DNA replication and damage repair. How its function is controlled remains an important question. Here, we show that the chromatin-bound PCNA protein is phosphorylated on Tyr 211, which is required for maintaining its function on chromatin and is dependent on the tyrosine kinase activity of EGF receptor (EGFR) in the nucleus. Phosphorylation on Tyr 211 by EGFR stabilizes chromatin-bound PCNA protein and associated functions. Consistently, increased PCNA Tyr 211 phosphorylation coincides with pronounced cell proliferation, and is better correlated with poor survival of breast cancer patients, as well as nuclear EGFR in tumours, than is the total PCNA level. These results identify a novel nuclear mechanism linking tyrosine kinase receptor function with the regulation of the PCNA sliding clamp. PMID- 17115033 TI - Hierarchical regulation of mitochondrion-dependent apoptosis by BCL-2 subfamilies. AB - Although the BCL-2 family constitutes a crucial checkpoint in apoptosis, the intricate interplay between these family members remains elusive. Here, we demonstrate that BIM and PUMA, similar to truncated BID (tBID), directly activate BAX-BAK to release cytochrome c. Conversely, anti-apoptotic BCL-2-BCL-X(L)-MCL-1 sequesters these 'activator' BH3-only molecules into stable complexes, thus preventing the activation of BAX-BAK. Extensive mutagenesis of BAX-BAK indicates that their activity is not kept in check by BCL-2-BCL-X(L)-MCL-1. Anti-apoptotic BCL-2 members are differentially inactivated by the remaining 'inactivator' BH3 only molecules including BAD, NOXA, BMF, BIK/BLK and HRK/DP5. BAD displaces tBID, BIM or PUMA from BCL-2-BCL-X(L) to activate BAX-BAK, whereas NOXA specifically antagonizes MCL-1. Coexpression of BAD and NOXA killed wild-type but not Bax, Bak doubly deficient cells or Puma deficient cells with Bim knockdown, indicating that activator BH3-only molecules function downstream of inactivator BH3-only molecules to activate BAX-BAK. Our data establish a hierarchical regulation of mitochondrion-dependent apoptosis by various BCL-2 subfamilies. PMID- 17115034 TI - Forward chemical genetic approach identifies new role for GAPDH in insulin signaling. AB - Insulin and insulin-like growth factor have an essential role in growth, development and the maintenance of metabolic homeostasis, including glucose uptake from the bloodstream. Researchers have identified mutations in insulin receptors that cause severe insulin resistance, and a temperature-sensitive daf-2 (a gene encoding an insulin receptor-like protein) mutant in Caenorhabditis elegans has served as an insulin resistance model. Here we report a forward chemical genetic approach with a tagged library that we used to identify a small molecule, GAPDH segregator (GAPDS), that suppresses the dauer formation induced by the daf-2 mutant. Like insulin, GAPDS increased both glucose uptake and the concentration of phosphatidylinositol-3,4,5-trisphosphate (PtdIns(3,4,5)P(3)) in mammalian preadipocytes. Using affinity matrices and RNA interference, we identified glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a GAPDS target. We discovered that GAPDH stimulates phosphatase activity against not only PtdIns(3,4,5)P(3) but also PtdIns(4,5)P(2). These results suggest that GAPDH is both an active regulator in the phosphoinositide-mediated signaling pathway and a potential new target for insulin resistance treatment. PMID- 17115035 TI - Sensitive and specific method for detecting G protein-coupled receptor mRNAs. AB - G protein-coupled receptors (GPCRs) mediate effects of extracellular signaling molecules in all the body's cells. These receptors are encoded by scarce mRNAs; therefore, detecting their transcripts with conventional microarrays is difficult. We present a method based on multiplex PCR and array detection of amplicons to assay GPCR gene expression with as little as 1 mug of total RNA, and using it, we profiled three human bone marrow stromal cell (BMSC) lines. PMID- 17115036 TI - fruitless regulates aggression and dominance in Drosophila. AB - When competing for resources, two Drosophila melanogaster flies of the same sex fight each other. Males and females fight with distinctly different styles, and males but not females establish dominance relationships. Here we show that sex specific splicing of the fruitless gene plays a critical role in determining who and how a fly fights, and whether a dominance relationship forms. PMID- 17115037 TI - SYD-2 Liprin-alpha organizes presynaptic active zone formation through ELKS. AB - A central event in synapse development is formation of the presynaptic active zone in response to positional cues. Three active zone proteins, RIM, ELKS (also known as ERC or CAST) and Liprin-alpha, bind each other and are implicated in linking active zone formation to synaptic vesicle release. Loss of function in Caenorhabditis elegans syd-2 Liprin-alpha alters the size of presynaptic specializations and disrupts synaptic vesicle accumulation. Here we report that a missense mutation in the coiled-coil domain of SYD-2 causes a gain of function. In HSN synapses, the syd-2(gf) mutation promotes synapse formation in the absence of syd-1, which is essential for HSN synapse formation. syd-2(gf) also partially suppresses the synaptogenesis defects in syg-1 and syg-2 mutants. The activity of syd-2(gf) requires elks-1, an ELKS homolog; but not unc-10, a RIM homolog. The mutant SYD-2 shows increased association with ELKS. These results establish a functional dependency for assembly of the presynaptic active zone in which SYD-2 plays a key role. PMID- 17115038 TI - Cochlear efferent feedback balances interaural sensitivity. AB - Neurons in the lateral superior olive (LSO) compute sound location based on differences in interaural intensity, coded in ascending signals from the two cochleas. Unilateral destruction of the neuronal feedback from the LSO to the cochlea, the lateral olivocochlear efferents, disrupted the normal interaural correlation in response amplitudes to sounds of equal intensity. Thus, lateral olivocochlear feedback maintains the binaural balance in neural excitability required for accurate localization of sounds in space. PMID- 17115039 TI - Hierarchical assembly of presynaptic components in defined C. elegans synapses. AB - The presynaptic regions of axons accumulate synaptic vesicles, active zone proteins and periactive zone proteins. However, the rules for orderly recruitment of presynaptic components are not well understood. We systematically examined molecular mechanisms of presynaptic development in egg-laying synapses of Caenorhabditis elegans, demonstrating that two scaffolding molecules, SYD-1 and SYD-2, have key roles in presynaptic assembly. SYD-2 (liprin-alpha) was previously shown to regulate the size and the shape of active zones. We now show that in syd-1 and syd-2 mutants, synaptic vesicles and numerous other presynaptic proteins fail to accumulate at presynaptic sites. SYD-1 and SYD-2 function cell autonomously at presynaptic terminals, downstream of synaptic specificity molecule SYG-1. SYD-1 is likely to act upstream of SYD-2 to positively regulate its synaptic assembly activity. These data imply a hierarchical organization of presynaptic assembly, in which transmembrane specificity molecules initiate synaptogenesis by recruiting a few key scaffolding proteins, which in turn assemble other presynaptic components. PMID- 17115040 TI - The P2Y12 receptor regulates microglial activation by extracellular nucleotides. AB - Microglia are primary immune sentinels of the CNS. Following injury, these cells migrate or extend processes toward sites of tissue damage. CNS injury is accompanied by release of nucleotides, serving as signals for microglial activation or chemotaxis. Microglia express several purinoceptors, including a G(i)-coupled subtype that has been implicated in ATP- and ADP-mediated migration in vitro. Here we show that microglia from mice lacking G(i)-coupled P2Y(12) receptors exhibit normal baseline motility but are unable to polarize, migrate or extend processes toward nucleotides in vitro or in vivo. Microglia in P2ry(12)(-/ ) mice show significantly diminished directional branch extension toward sites of cortical damage in the living mouse. Moreover, P2Y(12) expression is robust in the 'resting' state, but dramatically reduced after microglial activation. These results imply that P2Y(12) is a primary site at which nucleotides act to induce microglial chemotaxis at early stages of the response to local CNS injury. PMID- 17115041 TI - Dynorphin A activates bradykinin receptors to maintain neuropathic pain. AB - Dynorphin A is an endogenous opioid peptide that produces non-opioid receptor mediated neural excitation. Here we demonstrate that dynorphin induces calcium influx via voltage-sensitive calcium channels in sensory neurons by activating bradykinin receptors. This action of dynorphin at bradykinin receptors is distinct from the primary signaling pathway activated by bradykinin and underlies the hyperalgesia produced by pharmacological administration of dynorphin by the spinal route in rats and mice. Blockade of spinal B1 or B2 receptor also reverses persistent neuropathic pain but only when there is sustained elevation of endogenous spinal dynorphin, which is required for maintenance of neuropathic pain. These data reveal a mechanism for endogenous dynorphin to promote pain through its agonist action at bradykinin receptors and suggest new avenues for therapeutic intervention. PMID- 17115042 TI - Propagating waves mediate information transfer in the motor cortex. AB - High-frequency oscillations in the beta range (10-45 Hz) are most active in motor cortex during motor preparation and are postulated to reflect the steady postural state or global attentive state of the animal. By simultaneously recording multiple local field potential signals across the primary motor and dorsal premotor cortices of monkeys (Macaca mulatta) trained to perform an instructed delay reaching task, we found that these oscillations propagated as waves across the surface of the motor cortex along dominant spatial axes characteristic of the local circuitry of the motor cortex. Moreover, we found that information about the visual target to be reached was encoded in terms of both latency and amplitude of evoked waves at a time when the field phase-locked with respect to the target onset. These findings suggest that high-frequency oscillations may subserve intra- and inter-cortical information transfer during movement preparation and execution. PMID- 17115043 TI - Cannabinoids reveal importance of spike timing coordination in hippocampal function. AB - Cannabinoids impair hippocampus-dependent memory in both humans and animals, but the network mechanisms responsible for this effect are unknown. Here we show that the cannabinoids Delta(9)-tetrahydrocannabinol and CP55940 decreased the power of theta, gamma and ripple oscillations in the hippocampus of head-restrained and freely moving rats. These effects were blocked by a CB1 antagonist. The decrease in theta power correlated with memory impairment in a hippocampus-dependent task. By simultaneously recording from large populations of single units, we found that CP55940 severely disrupted the temporal coordination of cell assemblies in short time windows (<100 ms) yet only marginally affected population firing rates of pyramidal cells and interneurons. The decreased power of local field potential oscillations correlated with reduced temporal synchrony but not with firing rate changes. We hypothesize that reduced spike timing coordination and the associated impairment of physiological oscillations are responsible for cannabinoid-induced memory deficits. PMID- 17115044 TI - Dyslexia and the failure to form a perceptual anchor. AB - In a large subgroup of dyslexic individuals (D-LDs), reading difficulties are part of a broader learning and language disability. Recent studies indicate that D-LDs perform poorly in many psychoacoustic tasks compared with individuals with normal reading ability. We found that D-LDs perform as well as normal readers in speech perception in noise and in a difficult tone comparison task. However, their performance did not improve when these same tasks were performed with a smaller stimulus set. In contrast to normal readers, they did not benefit from stimulus-specific repetitions, suggesting that they have difficulties forming perceptual anchors. These findings are inconsistent with previously suggested static models of dyslexia. Instead, we propose that D-LDs' core deficit is a general difficulty in dynamically constructing stimulus-specific predictions, deriving from deficient stimulus-specific adaptation mechanisms. This hypothesis provides a direct link between D-LDs' high-level difficulties and mechanisms at the level of specific neuronal circuits. PMID- 17115045 TI - Simple fall-off pattern of correlated neural activity in the developing lateral geniculate nucleus. AB - Activity-dependent models for cortical simple-cell receptive field development predict specific patterns of correlated neural activity within the visual pathway, such as a Mexican hat-shaped pattern of correlated activity in the lateral geniculate nucleus (LGN). However, such activity patterns have yet to be experimentally demonstrated. We performed multielectrode recordings in the LGN of immature awake ferrets and found simple fall-off-shaped, rather than Mexican hat shaped, patterns of correlated activity. A weak surround in the LGN neuron's receptive field and the statistics of the input contributed to this pattern of correlated activity. Computer simulation of cortical receptive field development incorporating the experimentally observed activity patterns demonstrated that a simple-cell receptive field emerges when a newly devised 'split' constraint on synaptic growth is combined with Hebbian synaptic modification rules. Thus, given certain developmental constraints on synaptic plasticity, patterns of correlated activity within the LGN are compatible with Hebbian models of simple-cell receptive field development. PMID- 17115046 TI - Microbial translocation is a cause of systemic immune activation in chronic HIV infection. AB - Chronic activation of the immune system is a hallmark of progressive HIV infection and better predicts disease outcome than plasma viral load, yet its etiology remains obscure. Here we show that circulating microbial products, probably derived from the gastrointestinal tract, are a cause of HIV-related systemic immune activation. Circulating lipopolysaccharide, which we used as an indicator of microbial translocation, was significantly increased in chronically HIV-infected individuals and in simian immunodeficiency virus (SIV)-infected rhesus macaques (P 1.5 million SNPs. Some differences were simple insertions and deletions, but in regions containing CNVs, segmental duplication and repetitive DNA, they were more complex. Our results uncover substantial undescribed variation in humans, highlighting the need for comprehensive annotation strategies to fully interpret genome scanning and personalized sequencing projects. PMID- 17115058 TI - Conservation of hotspots for recombination in low-copy repeats associated with the NF1 microdeletion. AB - Several large-scale studies of human genetic variation have provided insights into processes such as recombination that have shaped human diversity. However, regions such as low-copy repeats (LCRs) have proven difficult to characterize, hindering efforts to understand the processes operating in these regions. We present a detailed study of genetic variation and underlying recombination processes in two copies of an LCR (NF1REPa and NF1REPc) on chromosome 17 involved in the generation of NF1 microdeletions and in a third copy (REP19) on chromosome 19 from which the others originated over 6.7 million years ago. We find evidence for shared hotspots of recombination among the LCRs. REP19 seems to contain hotspots in the same place as the nonallelic recombination hotspots in NF1REPa and NF1REPc. This apparent conservation of patterns of recombination hotspots in moderately diverged paralogous regions contrasts with recent evidence that these patterns are not conserved in less-diverged orthologous regions of chimpanzees. PMID- 17115059 TI - In germ cells of mouse embryonic ovaries, the decision to enter meiosis precedes premeiotic DNA replication. AB - The transition from mitosis to meiosis is a defining juncture in the life cycle of sexually reproducing organisms. In yeast, the decision to enter meiosis is made before the single round of DNA replication that precedes the two meiotic divisions. We present genetic evidence of an analogous decision point in the germ line of a multicellular organism. The mouse Stra8 gene is expressed in germ cells of embryonic ovaries, where meiosis is initiated, but not in those of embryonic testes, where meiosis does not begin until after birth. Here we report that in female embryos lacking Stra8 gene function, the early, mitotic development of germ cells is normal, but these cells then fail to undergo premeiotic DNA replication, meiotic chromosome condensation, cohesion, synapsis and recombination. Combined with previous findings, these genetic data suggest that active differentiation of ovarian germ cells commences at a regulatory point upstream of premeiotic DNA replication. PMID- 17115060 TI - Hematopoietic progenitor kinase 1 negatively regulates T cell receptor signaling and T cell-mediated immune responses. AB - HPK1 is a Ste20-related serine-threonine kinase that inducibly associates with the adaptors SLP-76 and Gads after T cell receptor (TCR) signaling. Here, HPK1 deficiency resulted in enhanced TCR-induced phosphorylation of SLP-76, phospholipase C-gamma1 and the kinase Erk, more-persistent calcium flux, and increased production of cytokines and antigen-specific antibodies. Furthermore, HPK1-deficient mice were more susceptible to experimental autoimmune encephalomyelitis. Although the interaction between SLP-76 and Gads was unaffected, the inducible association of SLP-76 with 14-3-3tau (a phosphorylated serine-binding protein and negative regulator of TCR signaling) was reduced in HPK1-deficient T cells after TCR stimulation. HPK1 phosphorylated SLP-76 and induced the interaction of SLP-76 with 14-3-3tau. Our results indicate that HPK1 negatively regulates TCR signaling and T cell-mediated immune responses. PMID- 17115061 TI - Hyaluronate and risperidone for hemorrhagic cystitis. PMID- 17115063 TI - Significance of additional high-dose cytarabine in combination with cyclophosphamide plus total body irradiation regimen for allogeneic stem cell transplantation. AB - The combination of cyclophosphamide (CY) and total body irradiation (TBI) has been used as a standard conditioning regimen for allogeneic transplantation. Several studies showed an advantage of adding high-dose cytarabine (HDCA) to this regimen. To clarify the significance of additional HDCA, we conducted a retrospective multicenter study and compared the clinical results of these two regimens. From June 1985 to March 2003, 219 patients with hematological malignancies underwent allogeneic transplantation after conditioning with CY+TBI 12Gy (n=73) or CA+CY+TBI 12Gy (n=146). Engraftment, overall survival, transplant related mortality (TRM), relapse rate and incidence of graft-versus-host disease (GVHD) were compared according to risks and donors. Addition of HDCA had no impact on the relapse rate in all subgroups, and it was associated with lower TRM among standard-risk patients after related transplantation, and with higher TRM and worse survival among standard-risk patients after unrelated transplantation. The incidence of acute GVHD was not significantly different between the two regimens, and HDCA resulted in a higher incidence of chronic GVHD among standard risk patients after related transplantation. In summary, addition of HDCA is not beneficial for high-risk patients, and is not recommended for standard-risk patients receiving unrelated transplantation. PMID- 17115062 TI - LACE-conditioned autologous stem cell transplantation for relapsed or refractory Hodgkin's lymphoma: treatment outcome and risk factor analysis in 67 patients from a single centre. AB - High-dose chemotherapy followed by autologous stem cell transplantation (ASCT) is a recognized treatment option for patients with relapsed Hodgkin's lymphoma. We have analysed 67 patients who underwent ASCT after LACE (lomustine (CCNU), cytarabine (Ara-C), cyclophosphamide, etoposide) conditioning for relapsed (n=61) or primary refractory (n=6) Hodgkin's lymphoma. The 100-day treatment-related mortality was 3%. With a median follow-up of 67 months (range 3.3-161.0) the probabilities of overall survival (OS) and progression-free survival (PFS) at 5 years were 68 and 64%, respectively. Probabilities for OS and PFS at 5 years for patients with chemosensitive relapse (n=40) were 81 and 78% versus 50 and 35%, respectively, for patients (n=27) with chemoresistant relapse (P=0.012 for OS, P=0.002 for PFS). In multivariate analysis mixed cellularity classical or lymphocyte-depleted classical histology subtype and haemoglobin level of 10 g/dl or less at the time of ASCT were identified as risk factors for worse OS, whereas stage III or IV disease at diagnosis and disease status at ASCT other than complete or partial remission predicted inferior PFS. LACE followed by ASCT is an effective treatment for the majority of patients with chemosensitive relapsed Hodgkin's lymphoma and a proportion of chemorefractory patients also benefit. PMID- 17115064 TI - Submyeloablative cord blood transplantation corrects clinical defects seen in IPEX syndrome. PMID- 17115065 TI - Conditioning with fludarabine alone and allogeneic transplantation as a successful rescue therapy for persistent, severe iatrogenic aplasia after relapse of acute myeloid leukemia. PMID- 17115066 TI - Short-term methotrexate could reduce early immune reactions and improve outcomes in umbilical cord blood transplantation for adults. AB - Post transplant immune disorders are problematic in cord blood transplantation (CBT) for adult patients, and optimal prophylaxis has not been established. We investigated whether intensive graft-versus-host disease (GVHD) prophylaxis using short-term methotrexate (MTX) has a prognostic impact on CBT. Post-CBT immune reactions were classified according to time course as pre-engraftment immune reaction (PIR), engraftment syndrome (ES) or acute GVHD. Between March 2001 and November 2005, a total of 77 patients underwent CBT at eight transplantation centers. Median age was 48 years (range, 18-69 years). Preparative regimens comprised myeloablative (n=31) or reduced-intensity (n=46). Acute GVHD prophylaxis included cyclosporine alone (n=23), tacrolimus alone (n=12), cyclosporine plus MTX (n=17), tacrolimus plus short-term MTX (n=23) or cyclosporine plus methylprednisolone (n=2). Cumulative incidences of PIR, ES and grade II-IV GVHD were 36, 12 and 23%, respectively. Short-term MTX exerted significant favorable effects on post-CBT immune reactions (hazard ratio, 0.55; 95% confidence interval (95% CI), 0.31-0.98; P=0.04) in multivariate analysis. Overall survival rates for patients with and without short-term MTX at day 180 were 59% (95% CI, 42-73%) and 16% (95% CI, 6.6-30%) (P=0.0001), respectively. Short-term MTX could offer one optimal regimen to reduce immune reactions and improve outcomes in CBT. PMID- 17115067 TI - Study on the cyclic GMP-dependency of relaxations to endogenous and exogenous nitric oxide in the mouse gastrointestinal tract. AB - BACKGROUND AND PURPOSE: cGMP mediates nitrergic relaxations of intestinal smooth muscle, but several studies have indicated that cGMP-independent mechanisms may also be involved. We addressed this contention by studying the effect of ODQ and ns2028, specific inhibitors of soluble guanylate cyclase, on nitrergic relaxations of the mouse gut. EXPERIMENTAL APPROACH: Mouse gastric fundus and small intestinal muscle preparations were mounted in organ baths to study relaxations to exogenous NO, NO donors and electrical field stimulation (EFS) of enteric nerves. KEY RESULTS: In gastric fundus longitudinal muscle strips, ODQ and NS2028 abolished the L-nitroarginine-sensitive relaxations to EFS and the relaxations to NO and NO donors, glyceryl trinitrate (GTN), SIN-1 and sodium nitroprusside (SNP). EFS of intestinal segments and muscle strips showed L nitroarginine-resistant relaxations, which were abolished by the purinoceptor blocker suramin. In the presence of suramin, ODQ and NS2028 abolished all relaxations to EFS in intestinal segments and strips. ODQ and NS2028 abolished the relaxations to exogenous NO and to the NO donors GTN, SIN-1 and SNP in circular and longitudinal intestinal muscle strips. Intestinal segments showed residual relaxations to NO and GTN. CONCLUSIONS AND IMPLICATIONS: Our results indicate that relaxations to endogenous NO in the mouse gastric fundus and small intestine are completely dependent on cGMP. ODQ and NS2028 incompletely blocked nitrergic relaxations to exogenous NO in intact intestinal segments. However, it is unlikely that this is due to the involvement of cGMP-independent pathways because ODQ and NS2028 abolished all relaxations to endogenous and exogenous NO in intestinal muscle strips. PMID- 17115068 TI - Beyond raloxifene for the prevention of osteoporosis and breast cancer. AB - Selective oestrogen receptor modulators (SERMs) can build bone in the postmenopausal woman and lower circulating cholesterol. These oestrogen-like properties contrast with the anti-oestrogenic properties observed in the breast where SERMs inhibit the oestrogen-mediated development and growth of ER positive breast cancers. The two clinically useful SERMs, tamoxifen and its chemical cousin raloxifene, are currently used successfully either for the treatment and prevention of breast cancer (tamoxifen) or the treatment and prevention of osteoporosis (raloxifene). However, raloxifene has the beneficial side-effect of breast cancer prevention. These multifunction medicines provide proof of concept that novel molecules can be selectively targeted to diseases mediated by the endocrine system. PMID- 17115069 TI - Alpha(2A)-adrenoceptors regulate sympathetic transmitter release in mice kidneys. AB - BACKGROUND AND PURPOSE: In the present study, a rodent model was used to investigate whether the alpha(2A)-adrenoceptor (alpha(2A)) represents the presynaptic autoinhibitory receptor regulating sympathetic transmitter release in the kidney. Moreover, the potential role of alpha(2A) as a heteroceptor regulating adenosine triphosphate (ATP) release was tested. EXPERIMENTAL APPROACH: Kidneys from wild-type (WT) and alpha(2A)-knockout (KO) mice were isolated and perfused. Renal nerves were stimulated with platinum-electrodes. Endogenously released noradrenaline (NA) was measured by HPLC. The perfusion pressure was monitored continuously. KEY RESULTS: Renal nerve stimulation (RNS) induced a frequency (1,2,5,7.5,10,15 Hz)-dependent release of NA in WT mice (994+/-373, 2355+/-541, 6375+/-950, 11626+/-1818, 19138+/-2001 pg NA g(-1) kidney (means+/-s.e.m.)). There was a 2.7-fold (5 Hz) increase of NA release in alpha(2A)-KO mice. In WT animals alpha-adrenoceptor blockade by phentolamine increased RNS-induced NA release in a concentration-dependent manner up to 350% of control. No facilitation by phentolamine was observed in alpha(2A)-KO mice. Pressor responses to 1 Hz and 2 Hz were resistant to alpha(1)-adrenoceptor blockade (0.03 microM prazosin) but abolished by P(2) receptor blockade (5 microM PPADS). Blockade of alpha(2)-adrenoceptors (1 microM rauwolscine) increased these purinergic pressor responses to 296+/-112% (1 Hz) in WT but not in alpha(2A)-KO mice. Exogenous ATP (100 microM) increased basal but not RNS-induced NA release. CONCLUSIONS AND IMPLICATIONS: alpha(2A)-Adrenoceptor-activation inhibits NA and ATP release from renal sympathetic nerves. Pressor responses to RNS at higher stimulation frequencies (>2 Hz) are mediated by NA. At lower frequencies neuronally released ATP seems to be the predominant transmitter mediating renovascular resistance. PMID- 17115070 TI - Biological activities of a novel selective oestrogen receptor modulator derived from raloxifene (Y134). AB - BACKGROUND AND PURPOSE: Selective oestrogen receptor (ER) modulators (SERMs) are of great value in the treatment of breast cancer and osteoporosis. The aim of this study was to characterize pharmacologically a new class of SERMs synthesized based on the core structure of raloxifene. EXPERIMENTAL APPROACH: Competitive receptor binding and luciferase-based reporter methods were used to study the bioactivities of raloxifene analogues, followed by efficacy determination in breast cancer cell proliferation assay. ER antagonist effects were investigated in female rats by measuring uterine and mammary gland growth, using wet weight, BrdU incorporation and terminal end bud (TEB) as indicators. KEY RESULTS: Five analogues, belonging to two different structural series and display higher binding affinities for ERalpha than ERbeta were functionally evaluated. One such analogue, Y134, exhibited potent antagonist activity at ERs in CV-1 cells cotransfected with plasmids containing ERalpha or ERbeta and oestrogen-response element-driven luciferase. The estimated IC(50) value was 0.52 nM for ERalpha and 2.94 nM for ERbeta, comparable to that of raloxifene. Little cytotoxicity was observed at Y134 concentrations below 10 microM. Y134 suppressed oestrogen stimulated proliferation of ER-positive human breast cancer MCF-7 and T47D cells. At an identical dose, administered to ovariectomized rats, Y134 was more effective than raloxifene at arresting oestrogen-induced outgrowth of TEB and mammary gland DNA synthesis, but their inhibitory effects on the uterus were comparable. CONCLUSIONS AND IMPLICATIONS: Y134 is a potent ER antagonist with better mammary gland selectivity than raloxifene and shows potential for development as a new SERM for therapeutic use. PMID- 17115071 TI - Regulation of OX1 orexin/hypocretin receptor-coupling to phospholipase C by Ca2+ influx. AB - BACKGROUND AND PURPOSE: Orexin (OX) receptors induce Ca2+ elevations via both receptor-operated Ca2+ channels (ROCs) and the "conventional" phospholipase C (PLC)-Ca2+ release-store-operated Ca2+ channel (SOC) pathways. In this study we assessed the ability of these different Ca2+ influx pathways to amplify OX1 receptor signalling to PLC in response to stimulation with the physiological ligand orexin-A. EXPERIMENTAL APPROACH: PLC activity was assessed in CHO cells stably expressing human OX1 receptors. KEY RESULTS: Inhibition of total Ca2+ influx by reduction of the extracellular [Ca2+] to 1 microM effectively inhibited the receptor-stimulated PLC activity at low orexin-A concentrations (by 93% at 1 nM), and this effect was gradually reduced by higher orexin-A concentrations. A similar but weaker inhibitory effect (84% at 1 nM) was obtained on depolarization to approximately 0 mV, which disrupts most of the driving force for Ca2+ entry. The inhibitor of the OX1 receptor-activated ROCs, tetraethylammonium chloride (TEA), was somewhat less effective than the reduction in extracellular [Ca2+] at inhibiting PLC activation, probably because it only partially blocks ROCs. The partial inhibitor of both ROCs and SOCs, Mg2+, and the SOC inhibitors, dextromethorphan, SKF-96365 (1-[beta-(3-(4-methoxyphenyl)propoxy)-4 methoxyphenethyl]-1H-imidazole HCL) and 2-APB (2-aminoethoxydiphenyl borate), inhibited PLC activity at low concentrations of orexin-A, but were not as effective as TEA. CONCLUSIONS AND IMPLICATIONS: Both ROCs and SOCs markedly amplify the OX(1) receptor-induced PLC response, but ROCs are more central for this response. These data indicate the crucial role of ROCs in orexin receptor signalling. PMID- 17115072 TI - Alpha(1A)-adrenoceptors mediate contractions to phenylephrine in rabbit penile arteries. AB - BACKGROUND AND PURPOSE: Maintained penile erection depends on the absence of alpha-adrenoceptor (alpha-AR) activation and so can be facilitated by alpha blockers. This study seeks the alpha(1)-AR subtypes involved in order to inform the pro-erectile consequences of subtype selective blockade. EXPERIMENTAL APPROACH: Wire myography was used with dorsal (nutritional supply) and cavernous (erectile inflow) penile arteries; standard alpha-AR-selective agonists and antagonists were employed to classify responses. KEY RESULTS: In both penile arteries noradrenaline (NA) and phenylephrine (PE, alpha(1)-AR agonist) caused concentration-dependent contractions. Sensitivity to NA was increased by NA uptake blockers, cocaine (3 microM) and corticosterone (30 microM). PE responses were antagonised by phentolamine (non-selective alpha-AR: dorsal pK(B) 8.00, cavernous 8.33), prazosin (non-subtype-selective alpha(1)-AR: dorsal 8.60, cavernous 8.41) and RS100329 (alpha(1A)-AR selective: dorsal 9.03, cavernous 8.80) but not by BMY7378 (alpha(1D)-AR selective: no effect at 1-100 nM) or Rec15/2615 (alpha(1B)-AR selective: no effect at 1-100 nM). Schild analysis was straightforward in cavernous artery, indicating that PE activates only alpha(1A) AR. In dorsal artery Schild slopes were low, though alpha(1A)-AR was still indicated. Analysis using UK 14,304 and rauwolscine indicated an alpha(2)-AR component in dorsal artery that may account for low slopes to alpha(1)-AR antagonists. CONCLUSIONS AND IMPLICATIONS: Penile arteries have a predominant, functional alpha(1A)-AR population with little evidence of other alpha(1)-AR subtypes. Dorsal arteries (nutritional supply) also have alpha(2)-ARs. Thus, alpha-AR blockers with affinity for alpha(1A)-AR or alpha(2)-AR would potentially have pro-erectile properties; the combination of these perhaps being most effective. This should inform the design of drugs to assist/avoid penile erection. PMID- 17115073 TI - The neuronal microRNA system. AB - A class of small, non-coding transcripts called microRNAs (miRNAs) that provide a crucial and pervasive layer of post-transcriptional gene regulation has recently emerged and become the focus of intense research. miRNAs are abundant in the nervous system, where they have key roles in development and are likely to be important mediators of plasticity. A highly conserved pathway of miRNA biogenesis is closely linked to the transport and translatability of mRNAs in neurons. Although there are nearly 500 known human miRNA sequences, each of only approximately 21 nucleotides, which bind to multiple mRNA targets, the accurate prediction of miRNA targets seems to lie just beyond our grasp. Nevertheless, the identification of such targets promises to provide new insights into many facets of neuronal function. PMID- 17115074 TI - High-conductance potassium channels of the SLO family. AB - High-conductance, 'big' potassium (BK) channels encoded by the Slo gene family are among the largest and most complex of the extended family of potassium channels. The family of SLO channels apparently evolved from voltage-dependent potassium channels, but acquired a large conserved carboxyl extension, which allows channel gating to be altered in response to the direct sensing of several different intracellular ions, and by other second-messenger systems, such as those activated following neurotransmitter binding to G-protein-coupled receptors (GPCRs). This versatility has been exploited to serve many cellular roles, both within and outside the nervous system. PMID- 17115075 TI - Axonal conduction and injury in multiple sclerosis: the role of sodium channels. AB - Multiple sclerosis (MS) is the most common cause of neurological disability in young adults. Recent studies have implicated specific sodium channel isoforms as having an important role in several aspects of the pathophysiology of MS, including the restoration of impulse conduction after demyelination, axonal degeneration and the mistuning of Purkinje neurons that leads to cerebellar dysfunction. By manipulating the activity of these channels or their expression, it might be possible to develop new therapeutic approaches that will prevent or limit disability in MS. PMID- 17115076 TI - The mirror neuron system and the consequences of its dysfunction. AB - The discovery of premotor and parietal cells known as mirror neurons in the macaque brain that fire not only when the animal is in action, but also when it observes others carrying out the same actions provides a plausible neurophysiological mechanism for a variety of important social behaviours, from imitation to empathy. Recent data also show that dysfunction of the mirror neuron system in humans might be a core deficit in autism, a socially isolating condition. Here, we review the neurophysiology of the mirror neuron system and its role in social cognition and discuss the clinical implications of mirror neuron dysfunction. PMID- 17115078 TI - The short-latency dopamine signal: a role in discovering novel actions? AB - An influential concept in contemporary computational neuroscience is the reward prediction error hypothesis of phasic dopaminergic function. It maintains that midbrain dopaminergic neurons signal the occurrence of unpredicted reward, which is used in appetitive learning to reinforce existing actions that most often lead to reward. However, the availability of limited afferent sensory processing and the precise timing of dopaminergic signals suggest that they might instead have a central role in identifying which aspects of context and behavioural output are crucial in causing unpredicted events. PMID- 17115077 TI - Towards multimodal atlases of the human brain. AB - Atlases of the human brain have an important impact on neuroscience. The emergence of ever more sophisticated imaging techniques, brain mapping methods and analytical strategies has the potential to revolutionize the concept of the brain atlas. Atlases can now combine data describing multiple aspects of brain structure or function at different scales from different subjects, yielding a truly integrative and comprehensive description of this organ. These integrative approaches have provided significant impetus for the human brain mapping initiatives, and have important applications in health and disease. PMID- 17115079 TI - Effect of Rho-associated kinase inhibition on actin cytoskeleton structure and calcium response in glioma C6 cells. AB - The role of actin cytoskeleton functional state in glioma C6 cell morphology and calcium signaling was investigated through modification of myosin II activity by blocking Rho-associated kinase with the specific inhibitor Y-27632. Treatment of glioma C6 cells with ROCK inhibitor resulted in actin cytoskeleton reorganization and also in the changed shape and distribution of mitochondria. Changes in the distribution of ER, the main calcium store in glioma C6 cells, were not visible. The inhibition of myosin II activity influences the first phase of calcium signaling evoked by agonist, and both phases of thapsigargin-evoked calcium response. We suggest that the observed increase in Ca2+ release from intracellular stores induced by IP3 formation as well as inhibition of SERCA ATPase is at least in part related to severely affected mitochondria. Enhancement of capacitative calcium entry evoked by thapsigargin is probably associated with the reorganization of the acto-myosin II system. ATP-induced calcium response presents no changes in the second phase. We observed that ATP stimulation of Y 27632 pretreated cells leads to immediate morphological rearrangement of glioma C6 cells. It is a consequence of actin cytoskeleton reorganization: formation of stress fibers and relocation of phosphorylated myosin II to actin filaments. It seems that the agonist-evoked strong calcium signal may be sufficient for myosin II activation and the stress fiber organization. This is the first work showing the dependence between the functional state of the acto-myosin II system and calcium signaling stressing the reversible character of this relationship. PMID- 17115080 TI - Thyroid hormones and their receptors in the regulation of cell proliferation. AB - In the present work, we have reviewed data showing that triiodothyronine and its nuclear receptors modify expression of different genes/proteins involved in cell cycle control beginning from growth factors (such as EGF and TGF-beta), to cell surface receptors (EGFR), as well as proteins acting at the cell membrane (Ras), various transcription factors (c-Fos, c-Myc, E2F1), cyclins, Cip/Kip family of cdk2 inhibitors, and p53 inhibitor Mdm2 (Table 1). We have shown how TRs are also able to modify the fate of a cell, thanks to their ability to form complexes with other transcription factors such as p53 - a key regulator of apoptosis and proliferation. Available data show that the function of thyroid hormones and of their receptors on cell proliferation is not homogenous. In fact, it strongly depends on the cell type, its developmental state (progenitor or differentiated), its patho-physiological state (normal or tumor cell), and the so-called 'cellular context'. Therefore, it is not possible to uniformly recommend T3 treatment or T3 depletion to stop or initiate proliferation of all cell types. Instead, a very individual and careful action should be considered. PMID- 17115081 TI - Evaluation and measurement of health-related quality of life for individuals with diabetes mellitus by Health Utilities Index Mark 3 (HUI3) system. AB - The Health Utilities Index Mark 3 (HUI3) is an important indicator when measuring the health-related quality of life (HRQOL) and assessing the burden of disease, especially for chronic conditions such as diabetes mellitus (DM). The objectives of this study were to evaluate the scores of HRQOL for respondents with DM to examine associations between overall HUI3 scores and eight component attributes, and various sociodemographic and lifestyle attributes, and by doing so, provide information to improve the HRQOL of individuals with diabetes. The study was based on the Canadian National Population Survey (NPHS) longitudinal data, from 1994-1995 to 2002-2003. We evaluated overall HUI3 scores and eight attributes (vision, hearing, speech, ambulation, dexterity, emotion, cognition, and pain) between respondents with and without diabetes in relation to demographic characteristics (age, sex, lifestyle, and socioeconomic status using ANCOVA[analysis of covariance]). Awareness of diabetes appeared to affect the HRQOL of older age groups more so than younger age groups (p < 0.01). Diabetes also appeared to have a greater impact on males' quality of life compared to females', and among individuals with single marital status and low socioeconomic status (p < 0.01). These findings add to what is known about cognitive representations and the self-regulation of diabetes as well as the relationships between cognitive representations of diabetes, HRQOL, and behavioral factors. In particular, results from this study suggest the need to address ways of reducing the burden of diabetes associated with health behaviours, and increasing the quality of life for the individuals with diabetes in Canada. PMID- 17115082 TI - Human development VI: supracellular morphogenesis. The origin of biological and cellular order. AB - Uninterrupted morphogenesis shows the informational potentials of biological organisms. Experimentally disturbed morphogenesis shows the compensational dynamics of the biological informational system, which is the rich informational redundancy. In this paper, we use these data to describe morphogenesis in terms of the development of supracellular levels of the organism, and we define complex epigenesis and supracellular differentiation. We review the phenomena of regeneration and induction of Hydra and amphibians, and the higher animal's informational needs for developing their complex nervous systems. We argue, also building on the NO-GO theorem for ontogenesis as chemistry, that the traditional chemical explanations of high-level informational events in ontogenesis, such as transmutation, regeneration, and induction, are insufficient. We analyze the informational dynamics of three embryonic compensatory reactions to different types of disturbances: (1) transmutations of the imaginal discs of insects, (2) regeneration after removal of embryonic tissue, and (3) embryonic induction, where two tissues that normally are separated experimentally are made to influence each other. We describe morphogenesis as a complex bifurcation, and the resulting morphological levels of the organism as organized in a fractal manner and supported by positional information. We suggest that some kind of real nonchemical phenomenon must be taking form in living organisms as an information carrying dynamic fractal field, causing morhogenesis and supporting the organism's morphology through time. We argue that only such a phenomenon that provides information-directed self-organization to the organism is able to explain the observed dynamic distribution of biological information through morphogenesis and the organism's ability to rejuvenate and heal. PMID- 17115083 TI - Human development VII: a spiral fractal model of fine structure of physical energy could explain central aspects of biological information, biological organization and biological creativity. AB - In this paper we have made a draft of a physical fractal essence of the universe, a sketch of a new cosmology, which we believe to lay at the root of our new holistic biological paradigm. We present the fractal roomy spiraled structures and the energy-rich dancing "infinite strings" or lines of the universe that our hypothesis is based upon. The geometric language of this cosmology is symbolic and both pre-mathematical and pre-philosophical. The symbols are both text and figures, and using these we step by step explain the new model that at least to some extent is able to explain the complex informational system behind morphogenesis, ontogenesis, regeneration and healing. We suggest that it is from this highly dynamic spiraled structure that organization of cells, organs, and the wholeness of the human being including consciousness emerge. The model of "dancing fractal spirals" carries many similarities to premodern cultures descriptions of the energy of the life and universe. Examples are the Native American shamanistic descriptions of their perception of energy and the old Indian Yogis descriptions of the life-energy within the body and outside. Similar ideas of energy and matter are found in the modern superstring theories. The model of the informational system of the organism gives new meaning to Bateson's definition of information: "A difference that makes a difference", and indicates how information-directed self-organization can exist on high structural levels in living organisms, giving birth to their subjectivity and consciousness. PMID- 17115084 TI - Human development VIII: a theory of "deep" quantum chemistry and cell consciousness: quantum chemistry controls genes and biochemistry to give cells and higher organisms consciousness and complex behavior. AB - Deep quantum chemistry is a theory of deeply structured quantum fields carrying the biological information of the cell, making it able to remember, intend, represent the inner and outer world for comparison, understand what it "sees", and make choices on its structure, form, behavior and division. We suggest that deep quantum chemistry gives the cell consciousness and all the qualities and abilities related to consciousness. We use geometric symbolism, which is a pre mathematical and philosophical approach to problems that cannot yet be handled mathematically. Using Occam's razor we have started with the simplest model that works; we presume this to be a many-dimensional, spiral fractal. We suggest that all the electrons of the large biological molecules' orbitals make one huge "cell orbital", which is structured according to the spiral fractal nature of quantum fields. Consciousness of single cells, multi cellular structures as e.g. organs, multi-cellular organisms and multi-individual colonies (like ants) and human societies can thus be explained by deep quantum chemistry. When biochemical activity is strictly controlled by the quantum-mechanical super-orbital of the cell, this orbital can deliver energetic quanta as biological information, distributed through many fractal levels of the cell to guide form and behavior of an individual single or a multi-cellular organism. The top level of information is the consciousness of the cell or organism, which controls all the biochemical processes. By this speculative work inspired by Penrose and Hameroff we hope to inspire other researchers to formulate more strict and mathematically correct hypothesis on the complex and coherence nature of matter, life and consciousness. PMID- 17115085 TI - Human Development IX: a model of the wholeness of man, his consciousness, and collective consciousness. AB - In this paper we look at the rational and the emotional interpretation of reality in the human brain and being, and discuss the representation of the brain-mind (ego), the body-mind (Id), and the outer world in the human wholeness (the I or "soul"). Based on this we discuss a number of factors including the coherence between perception, attention and consciousness, and the relation between thought, fantasies, visions and dreams. We discuss and explain concepts as intent, will, morals and ethics. The Jungian concept of the human collective conscious and unconscious is also analyzed. We also hypothesis on the nature of intuition and consider the source of religious experience of man. These phenomena are explained based on the concept of deep quantum chemistry and infinite dancing fractal spirals making up the energetic backbone of the world. In this paper we consider man as a real wholeness and debate the concepts of subjectivity, consciousness and intent that can be deduced from such a perspective. PMID- 17115086 TI - Anterograde and retrograde effects of benzodiazepines on memory. AB - Benzodiazepines are known as "acquisition-impairing" molecules, and their effects on anterograde memory processes are well described. In contrast, the impact of benzodiazepines on retrograde memory and, more particularly, on retrieval processes, is only marginally studied. This mini-review provides an overlook of the main studies evidencing an effect of benzodiazepines on retrograde memory, both in humans and animals, with special emphasis on retrieval processes. The conditions for the emergence of the benzodiazepine-induced retrieval impairments are also discussed. PMID- 17115087 TI - Effectiveness of the Tier 1 Program of the Project P.A.T.H.S.: preliminary objective and subjective outcome evaluation findings. AB - There are two tiers of programs in the Project P.A.T.H.S. (Positive Adolescent Training through Holistic Social Programmes). In the Tier 1 Program, teaching units based on different positive youth development constructs are covered. Pre- and post-test data utilizing the Chinese Positive Youth Development Scale (CPYDS) and post-test subjective outcome evaluation data were collected from 546 students who participated in the 20 h Tier 1 Program of the P.A.T.H.S. Project. Results showed that high proportions of the respondents had positive perceptions of the program and the instructors, with 85.3% of the respondents regarding the program as helpful to them. Positive changes in the program participants in many measures of positive youth development were also observed. Although there were some increases in problem behavior in some areas, adolescent problem behavior was generally stable. The present study provides preliminary support for the effectiveness of the Tier 1 Program of the Project P.A.T.H.S. in Hong Kong. PMID- 17115088 TI - [Myxoma of the middle ear-a rare cause of facial palsy]. AB - In case of the co-occurrence of facial palsy and inflammation-like symptoms of the same ear, the differential diagnosis is focused on viral (herpes zoster) or bacterial diseases. We report a patient for whom the surgical exploration of the middle ear revealed a benign tumor: a myxoma. These neoplasias are rare tumors in the head and neck region. The typical tumor site is the atrium of heart. In the ear, the tumor grows slowly and remains asymptomatic unless it irritates structures such as the facial nerve or the vestibular organ. Histologically, the tumor presents a "myxoid" matrix that is rich in acid mucopolysaccarides. The treatment of choice is complete surgical resection. Using the case presented, we discuss the causality between the tumor and the facial palsy, although during the operation the bony canal of the nerve was found to be intact. In any cases with clinically and radiologically unclear findings of the ear in connection with facial palsy, surgical exposure should be considered. PMID- 17115089 TI - [Low frequency fluctuating hearing loss without labyrinthine vertigo--a genuine disease? A follow up study after 4 and 10 years]. AB - BACKGROUND: Besides the typical attacks of dizziness, recurrent low-frequency sensory hearing loss--together with mostly low-frequency tinnitus--is also a characteristic sign of Meniere's disease. It is therefore often assumed to be a prodromal sign of Meniere's disease, even without dizziness. METHODS: During our longitudinal study, which was started in 1995, we reported that there were 81 patients with recurrent low-frequency hearing loss with no initial vertigo in the first suspense year of 1999. After a further 6 years, we investigated 46 (57%) of these original 81 patients in the second suspense year, 2005, for new components of vertigo, with the questions of development of Meniere's disease and further development of the patients' hearing ability and psychic situation in mind. RESULTS: In all, 12 (26%) of the 81 former patients suffered from vertigo, but only 4 (9%) had developed the typical signs of full-blown Meniere's disease with the typical labyrinthine vertigo. Of the 12 patients who suffered from vertigo, 6 (13% of the 81 with vertigo) were diagnosed with psychogenic vertigo, 1 (1%) suffered from benign and treatable paroxysmal positional vertigo and 1 (1%) had developed vertigo after acoustic neurinoma surgery. CONCLUSIONS: We conclude from our observations that, although almost every patient with Meniere's disease suffers from recurrent low-frequency hearing loss, only a few patients with recurrent hearing loss develop Meniere's disease. However, many patients with low frequency sensory hearing loss develop anxiety leading to psychogenic dizziness in fearful expectation of "imminent" Meniere's disease. We found that 26% of the patients had persisting bilateral normacusis in the low-frequency ranges, while 34% had unilateral hearing loss that was sufficiently severe to affect their lives and 39%, bilateral hearing loss; however, none of the patients became completely deaf. PMID- 17115090 TI - [Clinical experience with the hygienic reprocessing of rigid endoscopes lacking a working channel with reference to practicability in clinical routine]. AB - BACKGROUND: Contaminated medical products may be vectors for infections. A safe disinfection method for the optical instruments used by ENT specialists is undoubtedly needed. So far, a standard method that sufficiently covers the risk of infection for patients and medical staff alike, while serving the need of practicability in daily routine, has not been established. PATIENTS AND METHODS: In the ENT departments of the Mannheim and Kaiserslautern hospitals, a study on the practicability of disinfection rigid optical instruments lacking working channels was conducted using cleaning and disinfection automats. A total of 735 patients were examined (Hopkins 30 degrees and 70 degrees) and the endoscopes subsequently sterilized using a cleaning and disinfection automat. RESULTS: Each cleaning cycle took about 40 min. Examining 70-100 patients a day, a minimum of eight 70 degrees Hopkins endoscopes would be needed. CONCLUSION: The sterilization of medical products such as endoscopes is the best practice, but certain hindrances, such as loss of time and costs involved, complicate its general application. PMID- 17115091 TI - [Sinonasal sarcoidosis]. AB - BACKGROUND: The sinonasal system is rarely involved in the clinical picture of sarcoidosis. In the absence of pulmonary disease, sinonasal sarcoidosis is extremely rare. PATIENTS AND METHODS: Four patients with isolated sarcoidosis of the nose and the sinuses are reported; in one of these patients the histological evidence was found in the mucosa of the nasopharynx. RESULTS: None of these patients was found to have a pulmonary illness or any other extrapulmonary manifestation of sarcoidosis. Retrospectively, two patients fulfilled the more specific diagnostic criteria for sinonasal sarcoidosis reported in the literature. CONCLUSION: Sarcoidosis should be considered in the differential diagnosis of inflammatory sinonasal disease, especially if the mucosa shows signs of granulation, but it should also be considered when an incidental finding of rhinosinusitis is recorded. Rhinosurgical intervention seems to be an appropriate therapy in terms of improving the symptoms of the disease, despite the prolonged period of postoperative healing and the necessity for individual treatment with medication. Following histological confirmation of the diagnosis, adequate medication and further appropriate diagnostic procedures drawing on internal medicine are essential. PMID- 17115092 TI - [Chronic polypous rhinosinusitis: Genesis, clinical picture, therapy and relapse rate--a retrospective study]. AB - BACKGROUND: The prevalence of chronic polypous rhinosinusitis is high at 1-2%. In the present study, the predisposing factors for this condition, the clinical symptomatology, results of surgical interventions, significance of computed tomography diagnosis, as well as histology and post-operative therapy with steroids were investigated. METHODS AND RESULTS: A 15% stratified sample, made up of 194 post-paranasal sinus surgery patients of 5 surgical years was investigated. Above all, the eosinophil dominated type led to the formation of polyps. In the diagnosis of chronic polypous rhinosinusitis computed tomography is only sufficiently sensitive for the ethmoid bone. The complication rate was, with 4.6%, in the lower range of comparable pre-examinations. In addition, the 18% relapse rate in the patient population, of whom 49% had already been operated on once or several times, corresponds to a good result when compared at an international level. CONCLUSIONS: These data confirm that surgical experience and a consistent post-surgical treatment can reduce the relapse frequency of chronic polypous rhinosinusitis. This is indicated by the on average late occurrence of a relapse. PMID- 17115093 TI - [Posttraumatic amaurosis after complex frontobasal fracture. Differential diagnosis and therapy]. AB - BACKGROUND: Whether surgical nerve decompression is indicated for the treatment of posttraumatic reduced vision with optic nerve compression in the bony canal has been a subject of controversy for decades. On balance, the recent literature suggests that this procedure is indeed indicated, as a supplement to high-dosed cortisone therapy. The risk of surgery-related side effects is usually rated low in the literature. CASE REPORT: We report on a woman patient in whom craniocerebral trauma involved a fracture of the left optic canal with unilateral loss of vision. In the decompression operation, intraoperative symptoms gave rise to the suspicion of an arteriovenous fistula, which had not been revealed by computer tomography and which was seen as sufficient grounds for discontinuing the procedure. In addition to a carotid artery-sinus cavernosus fistula (CCF Barrow type A), subsequent angiography revealed a dissected aneurysm at the branching of the occluded ophthalmic artery. DISCUSSION AND CONCLUSIONS: In decisions on whether surgical relief of pressure on the optic canal is indicated after trauma-related visual loss, the possibility of secondary lesions near the tip of the orbita and the optic canal must be taken into account. These are not always revealed by computer tomography. The indications should be critically weighed up in each individual case, with additional imaging examinations, such as MR-angiography, CT-angiography, or conventional angiography, performed as needed. The options and indications for imaging are discussed. The procedure can by no means be rated as "minimally invasive", as is postulated by some authors. PMID- 17115094 TI - [Critical factors for subjective burden of mothers of children with developmental language disorders]. AB - BACKGROUND: In earlier studies, it was found that mothers of children with developmental language disorders had a higher incidence of anxiety and depression in comparison to the normal population. This study concentrated on the following questions: is the perceived stress, anxiety and depression of mothers with children with developmental language disorders higher than in the normal population, even for those mothers who are not exposed to any further stressors? and what environmental factors play an important role in the mental health of the mothers? MATERIALS AND METHODS: A total of 90 mothers (aged 32.9+/-4.7 years) of preschoolers diagnosed with a developmental language disorder, but with otherwise normal development, participated in the study. The mental health of the mothers was assessed using the Perceived Stress Scale (PSS) and the German version of the Hospital Anxiety and Depression Scale (HASD-D). Environmental factors were captured by a structured anamnestic procedure. Predictors of mental health were determined using multivariate regression analysis. RESULTS: The entire sample demonstrated significant levels of stress, anxiety and depression. In comparison to the standardized values, those mothers of children with developmental language disorders who were not exposed to any further stressors, were also shown to have significantly higher rates of stress and depression on the scales used, although not significantly higher levels of anxiety. A significant predictor of positive overall mental health of the mothers was the extent of adequate support in child rearing. CONCLUSIONS: Health professionals dealing with children with developmental language disorders must reckon with a considerable number of mothers whose mental health is negatively affected. When treating a child with a developmental language disorder, measures to improve the mother's state of well being should be considered, including counseling and self-help groups. PMID- 17115095 TI - Bilateral orbital emphysema and pneumocephalus as a result of accidental compressed air exposure. AB - Orbital emphysema is a rare condition in the absence of trauma or sinus disease. A 22-year-old man suffering from left orbital trauma due to sudden exposure to compressed air tube was admitted with severe pain in the left eye, swelling, and mild periorbital ecchymosis. Physical examination revealed a large conjunctival laceration in the left orbit. Multislice computed tomographic scanning of the head and orbits showed extensive radiolucencies consistent with the air in both orbits, more prominent in the left. There was also subcutaneous air in the left periorbital soft tissue extending through fronto-temporal and zygomatic areas. Air was also demonstrated adjacent to the left optic canal and within the subarachnoid space intracranially. There was no evidence of any orbital, paranasal sinus, or cranial fracture. Visual acuity was minimally decreased bilaterally. The conjunctiva was sutured under local anesthesia. After 3 weeks of follow-up, the patient completely recovered without visual loss. Bilateral orbital emphysema with pneumocephalus can occur from a high-pressure compressed air injury after unilateral conjunctival trauma without any evidence of fracture. PMID- 17115096 TI - Calcified fecolith--a rare cause of large bowel obstruction. AB - The authors present a rare case of colonic obstruction caused by a fecolith in a 59-year-old female who presented with symptoms suggestive of intestinal obstruction. The computed tomography (CT) confirmed the diagnosis. The patient was managed by laparotomy and colotomy to remove the fecolith. PMID- 17115097 TI - Jejunogastric intussusception: a case report with the review of literature. AB - Jejunogastric intussusception (JGI) is a rare but potentially lethal complication of gastrectomy or gastrojejunostomy. In the acute setting, early diagnosis and prompt surgical intervention are mandatory to avoid mortality. We present a case of JGI in a patient with a history of gastrojejunostomy who had increasing vomiting, hematemesis, and abdominal pain for 1 day. PMID- 17115098 TI - Massive pulmonary embolus with hemodynamic compromise: therapeutic options. AB - Recent advances in pharmacotherapeutics and interventional techniques have resulted in resurgence in interest regarding the optimal means of management of hemodynamically significant pulmonary embolic disease. However, these various techniques have many associated disadvantages, and it is of paramount importance that the responsible physician has a detailed understanding of each of these so that the appropriate intervention be requested and performed. By way of background discussion and relevant cases in point, we consider each of these management options in turn, with particular reference to the advantages, disadvantages, and application of each. PMID- 17115099 TI - Meta-analysis is no substitute for a comprehensive national registry. AB - Anti-tumor necrosis factor alpha (TNF-alpha) agents have become an established treatment option for rheumatoid arthritis (RA), but are not without risks. As TNF alpha has a role in tumour surveillance, anti-TNF-alpha blockade could potentially increase the risk of malignancy. Recent meta-analysis by Bongartz et al. (JAMA 295:2275-2285, 2006) reported an increase in the incidence of malignancy attributed to anti-TNF-alpha antibodies, and the information has quickly and uncritically reached several secondary sources with huge effects on public perception of risks related to anti-TNF-alpha therapy. In contrast, the results from the British Society for Rheumatology Biologics Register show that in clinical practice, anti-TNF-alpha therapy in RA does not appear to increase the risk of malignancy in those patients with low risk of malignancy. We discuss the issues emerging from these published data and suggest caution against giving weight to meta-analysis of short-term drug studies. PMID- 17115101 TI - Mononeuritis multiplex as a presenting feature of Wegener granulomatosis: a case report. AB - We report the case of a man presenting with clinical features of a mononeuritis multiplex, a perinuclear-ANCA (p-ANCA), and a renal biopsy suggestive of Wegener granulomatosis (WG). We wish to highlight this case as a learning point for clinicians as WG rarely presents in this form, and can be easily overlooked as a cause of mononeuritis multiplex. PMID- 17115100 TI - Central sensitization: a biopsychosocial explanation for chronic widespread pain in patients with fibromyalgia and chronic fatigue syndrome. AB - In addition to the debilitating fatigue, the majority of patients with chronic fatigue syndrome (CFS) experience chronic widespread pain. These pain complaints show the greatest overlap between CFS and fibromyalgia (FM). Although the literature provides evidence for central sensitization as cause for the musculoskeletal pain in FM, in CFS this evidence is currently lacking, despite the observed similarities in both diseases. The knowledge concerning the physiological mechanism of central sensitization, the pathophysiology and the pain processing in FM, and the knowledge on the pathophysiology of CFS lead to the hypothesis that central sensitization is also responsible for the sustaining pain complaints in CFS. This hypothesis is based on the hyperalgesia and allodynia reported in CFS, on the elevated concentrations of nitric oxide presented in the blood of CFS patients, on the typical personality styles seen in CFS and on the brain abnormalities shown on brain images. To examine the present hypothesis more research is required. Further investigations could use similar protocols to those already used in studies on pain in FM like, for example, studies on temporal summation, spatial summation, the role of psychosocial aspects in chronic pain, etc. PMID- 17115102 TI - Anaplastic thyroid carcinoma: expression profile of targets for therapy offers new insights for disease treatment. AB - BACKGROUND: Anaplastic thyroid cancer is an endocrine malignancy. Its rare and rapidly lethal disease course has made it challenging to study. Little is known regarding the expression by anaplastic tumors of molecular targets for new human anticancer agents that have been studied in the preclinical or clinical setting. The objective of this work was to evaluate the expression profile of anaplastic thyroid tumors for molecular targets for treatment. METHODS: Of the 94 cases of anaplastic thyroid cancers diagnosed and treated in British Columbia, Canada over a 20-year period (1984-2004), 32 cases (34%) had adequate archival tissue available for evaluation. A tissue microarray was constructed from these anaplastic thyroid tumors and immunohistochemistry was utilized to evaluate expression of 31 molecular markers. The markers evaluated were: epidermal growth factor receptor (EGFR), HER2, HER3, HER4, ER, PR, uPA-R, clusterin, E-cadherin, beta-catenin, AMF-R, c-kit, VEGF, ILK, aurora A, aurora B, aurora C, RET, CA-IX, IGF1-R, p53, MDM2, p21, Bcl-2, cyclin D1, cyclin E, p27, calcitonin, MIB-1, TTF 1, and thyroglobulin. RESULTS: A single tumor with strong calcitonin expression was identified as a poorly differentiated medullary carcinoma and excluded from the study cohort. The mean age of the anaplastic cohort was 66 years; 16 patients (51%) were females, and the median patient survival was 23 weeks. A wide range in molecular marker expression was observed by the anaplastic thyroid cancer tumors (0-100%). The therapeutic targets most frequently and most strongly overexpressed by the anaplastic tumors were: beta-catenin (41%), aurora A (41%), cyclin E (67%), cyclin D1 (77%), and EGFR (84%). CONCLUSIONS: Anaplastic thyroid tumors exhibit considerable derangement of their cell cycle and multiple signal transduction pathways that leads to uncontrolled cellular proliferation and the development of genomic instability. This report is the first to comprehensively evaluate a panel of molecular targets for therapy of anaplastic thyroid cancer and supports the development of clinical trials with agents such as cetuximab, small-molecule tyrosine kinase inhibitors, and aurora kinase inhibitors, which may offer new hope for individuals diagnosed with this fatal thyroid malignancy. PMID- 17115103 TI - Perioperative interstitial brachytherapy for soft tissue sarcomas: prognostic factors and long-term results of 155 patients. AB - BACKGROUND: The goal of this study was to evaluate the efficacy of temporary interstitial brachytherapy (BRT) for patients undergoing combined modality management of soft tissue sarcomas (STS). METHODS: From January 1990 to December 2003, 155 adults 18-88 years of age (median = 42 years) with STS who had received BRT as part of locoregional treatment were included in this review. Sixty-four percent were males. Sixty-nine percent had primary lesions. Sixty percent had lesions involving the lower extremities. Spindle cell sarcoma (28%) and synovial sarcoma (16%) were the most common histologic types and 51% had grade III lesions. Treatment included wide local excision of primary tumor with BRT with or without external beam radiotherapy (EBRT). RESULTS: After a median followup of 45 months, the local control (LC), disease-free survival (DFS), and overall survival (OS) for the entire cohort was 71%, 57%, and 73%, respectively. DFS was superior for superficial tumors compared with that for deep tumors (96% vs. 54%, P =.02). Patients with a tumor less than 5 cm had superior OS (88% vs. 63%, P =.05). Cumulative radiotherapy dose greater than 60 Gy had a significant positive impact on LC (P = .003), DFS (P =.003), and OS (P =.048). Subcutaneous fibrosis (21%) was the major complication. CONCLUSIONS: Temporary perioperative iridium-192 interstitial BRT with or without EBRT after function-preserving surgery results in satisfactory outcome in patients with STS. Both low dose rate and high dose rate BRT are equivalent in terms of disease control and complications when used alone or in combination with EBRT. BRT results in fewer complications compared with the combination of BRT and EBRT. PMID- 17115104 TI - Response of Bdellovibrio and like organisms (BALOs) to the migration of naturally occurring bacteria to chemoattractants. AB - A dual culture-based and non-culture-based approach was applied to characterize predator bacterial groups in surface water samples collected from Apalachicola Bay, Florida. Chemotaxis drop assays were performed on concentrated samples in an effort to isolate predator bacteria by their chemotactic ability. Yeast extract (YE) and casamino acids (CA) proved to be strong chemoattractants and resulted in three visibly distinct bands; however, dextrose, succinate, pyruvate, and concentrated cells of Vibrio parahaemolyticus P5 as prey did not elicit any response. The three distinct bands from YE and CA were separately collected to identify the chemotactic microbial assemblages. Plaque-forming unit assays from different chemotaxis bands with P5 as prey indicated 5- (CA) to 10-fold (YE) higher numbers of predator bacteria in the outermost chemotactic bands. Polymerase chain reaction-restriction fragment length polymorphism and 16S rDNA sequencing of clones from different chemotaxis bands resulted in identification of Pseudoalteromonas spp., Marinomonas spp., and Vibrio spp., with their numbers inversely proportional to the numbers of predators-i.e., Bdellovibrio spp. and Bacteriovorax spp-in the chemotaxis bands. This study indicates that predatorial bacteria potentially respond to high densities of microbial biomass in aquatic ecosystems and that chemotaxis drop assay may be an alternate culture-independent method to characterize predatorial bacterial guilds from the environment. PMID- 17115105 TI - Prevalence of shiga toxin-producing Escherichia coli in a diarrheagenic Tunisian population, and the report of isolating STEC O157:H7 in Tunis. AB - In Mellassine (a major city in the state of Tunis) and Ben Arous state (south east of Tunis), a total of 212 stool samples were collected from children and adults (symptomatic and asymptomatic groups) between November 2001 and November 2004. Three hundred and twenty-seven E. coli strains were isolated and studied, to look for shiga toxin-producing Escherichia coli (STEC) strains, which were further analysed to investigate and determine clonal relationship among Tunisian STEC strains isolated from different sources (diarrheal cases and food products). They were analysed to characterize their serotypes, virulence genes by PCR, cytotoxic effect on Vero cell, plasmid profiles, and pulsed-field gel electrophoresis (PFGE) patterns. Eleven isolates (10 nontypeable, one O157:H7) carried stx gene and shared Stx restriction fragment length polymorphism (RFLP) patterns (stx1 ( + ), stx2 ( + )). Seven of these strains were isolated from acute diarrheal cases, and four were isolated from a control group (among which the only isolated STEC O157:H7). Two of the STEC strains harboured both eae and ehxA genes. Analysis of the cytotoxic effect on Vero cells showed that a correlation exists between carrying stx1 ( + ), stx2 ( + ) genes and cytotoxicity. Also a correlation was noticed between STEC strains recovered from different sources regarding plasmid profiles and PFGE patterns. All stool samples positive for STEC were nonbloody. None of the STEC-positive patients developed severe diseases. These data demonstrate that although STEC is not a major cause of acute diarrhea in Tunis, it should not be overlooked. Measures should be taken to improve the detection and isolation of STEC from acute diarrheal cases as well as carriers. PMID- 17115106 TI - Anaerobic biodegradation of natural gas condensate can be stimulated by the addition of gasoline. AB - Biodegradation of a broad range of linear and branched alkanes, parent and alkyl alicyclic hydrocarbons, and benzene and alkyl-substituted benzenes was observed when sediment and groundwater samples collected from a gas condensate contaminated aquifer were incubated under methanogenic and especially under sulfate-reducing conditions, even though no exogenous nitrogen or phosphorus was added. This finding expands the range of hydrocarbon molecules known to undergo anaerobic decay and confirms that natural attenuation is an important process at this site. The addition of 1 mul of gasoline to the samples (approximately 10 ppm) had minimal impact on the biodegradation of saturated compounds, but substantially increased the diversity and extent of aromatic compounds undergoing transformation. We attribute this to the promotion or induction of biodegradation pathways in the indigenous microflora following the addition of the gasoline components. The promoting compounds are not precisely known, but may have been present in the initial condensate and reduced in concentration by various mechanisms (dissolution, biodegradation, etc.) such that their concentration in the aquifer fell below necessary levels. A variety of aromatic hydrocarbons would appear to be likely candidates. PMID- 17115107 TI - Risk for contralateral breast cancers in a population covered by mammography: effects of family history, age at diagnosis and histology. AB - BACKGROUND: Improved survival for breast cancer is increasing the likelihood of contralateral tumors. Mammographic screening is partially contributing to the survival advantage, while changing many aspects of breast cancer presentation, including age at diagnosis, histology and familial risk. As mammography has become widely used, it is important to quantify the risks for contralateral breast cancer in a population with a national access to mammographic screening service. METHODS: The nation-wide Swedish Family-Cancer Database was used to calculate risks for contralateral breast cancer between years 1990 (1993) and 2002. The standardized incidence ratio (SIR) measured the risk for contralateral breast compared to first breast cancer. RESULTS: The risks for contralateral breast cancer ranged between 1.85 and 3.79, and they tended to be higher when in situ cancer was diagnosed. Family history and early diagnosis of first cancer increased the risks for contralateral breast cancer, approximately equally for invasive and in situ cancers. The risk for contralateral in situ cancer was 9.01 following two independent invasive cancers. The risk for the same, concordant histology between the first and the contralateral cancer was higher than that for discordant histologies. The risks for concordant histologies were particularly high for mucinous (12.16), comedo (11.74) and lobular (5.06) tumors. When the first lobular cancer was diagnosed before age 45 years, the risk for contralateral lobular cancer was 32.20. CONCLUSION: In situ breast cancer poses an approximately equally high risk as invasive cancer. Family history and earlier age of onset are associated with high risks needing clinical attention. PMID- 17115108 TI - Influence of young age at diagnosis and family history of breast or ovarian cancer on breast cancer outcomes in a population-based cohort study. AB - PURPOSE: The objective of this study was to examine the association of: (i) diagnosis at age or =36 years was, respectively, 1.08 (95% CI: 0.99-1.18), 1.12 (95% CI: 1.03-1.23), 1.17 (95% CI: 1.06-1.29), and 1.12 (95% CI: 1.01-1.25), compared to women born to mothers aged < or =20 years (P for trend = 0.008). Similarly, advanced paternal age was associated with increased incidence of breast cancer (P for trend = 0.03), but the association disappeared when conditioning on maternal age. The positive association between maternal age and incidence of breast cancer was stronger for estrogen receptor-positive and progesterone receptor-positive tumors (P for trend = 0.003) than for tumors with both receptors negative (P for trend = 0.78), and was more consistent among postmenopausal women, women without a family history and women who were first born. CONCLUSION: Our findings support a modest positive association between maternal age and daughter's risk of breast cancer, possibly mediated by hormonal factors. PMID- 17115114 TI - Frequent loss of Dab2 protein and infrequent promoter hypermethylation in breast cancer. AB - Disabled-2 (Dab2), a putative tumor suppressor protein, is lost in 80-90% ovarian tumors and ovarian/breast cancer cell lines. The clinical significance of Dab2 protein in breast cancer remains yet unknown. Immunohistochemical analysis of Dab2 protein showed no detectable expression in 67/91 (74%) breast tumors, while all 10 normal tissues showed presence of Dab2 protein. We hypothesized that epigenetic silencing of Dab2 may account for loss of protein in breast cancer. Methylation of Dab2 exon 1, a putative promoter, was analyzed in six breast cancer cell lines and in 54 primary breast tumors by methylation specific PCR. Methylation was observed in MDA-MB-231 and MDA-MB-157 cells and in 6 of 54 (11%) primary breast tumors that also showed loss of Dab2 protein. Expression of Dab2 transcripts was detected in all cell lines except MDA-MB-157. However, none of these six cell lines showed detectable levels of Dab2 protein by western blotting, while non-malignant mammary epithelial cell line MCF 10A showed Dab2 protein expression. To our knowledge this is the first report showing low frequency of Dab2 (putative) promoter methylation (11%) in primary breast tumors. Frequent loss of Dab2 protein (74%) suggest that hypermethylation of Dab2 promoter may only be one of the mechanisms accounting for its loss in breast cancer. Further, in silico analysis of Dab2 3'-UTR revealed existence of miRNA complimentary to this region of the gene, suggesting microRNA mediated targeting of Dab2 mRNA might account for loss of the protein in breast cancer. PMID- 17115116 TI - Effects of resveratrol in inflammatory arthritis. AB - Nuclear factor kappa B (NF-kappaB), is a pivotal transcription factor involved in the activation of the TNF-alpha and IL-1beta genes. Activation of NF-kappaB in synovial cells is a feature seen in arthritis patients. Resveratrol, a polyphenolic, natural phytoalexin found with particularly high levels in grape skin and red wine is potent and specific inhibitor of TNF-alpha and IL-1beta induced NF-kappaB activation. We aimed to determine the in vivo effects of intra articular injections of resveratrol on cartilage and synovium in an experimental rabbit inflammatory arthritis model. MATERIALS AND METHODS: Arthritis was induced by intra-articular injection of three times of 50 mug lipopolysaccharide (LPS) at day 0, 4 and 8 at 4-day intervals into the knee joints of rabbits. To the test group, 10 muMol/kg resveratrol in the DMSO was injected in the knees at day 0 and then it was continued once daily for 2 weeks. To the control group the same time and amount of DMSO was injected the knees of rabbits. All rabbits were killed 1 week after the last injection and cartilage tissue and synovium were evaluated with semiquantitative scoring histologically. RESULTS: According to control group in the resveratrol group, significantly decreased cartilage destruction was determined by H&E staining (p = 0.04). Loss of matrix proteoglycan content in the cartilage was much lower, as determined by safranin O staining (p = 0.03). We also observed marked synovial inflammation after intra-articular injection to control knees, but not in the resveratrol treated group knees (p = 0.01). CONCLUSION: This study suggests that intra-articular injection of resveratrol may protect cartilage against the development of experimentally induced IA. PMID- 17115117 TI - Proinsulin C-peptide and insulin: Limited pattern similarities of interest in inter-peptide interactions but no C-peptide effect on insulin and IGF-1 receptor signaling. AB - The recently reported influence of proinsulin C-peptide on insulin prompted us to examine structural features of the C-peptide. Four sets of limited pattern similarities towards inter-chain end regions of insulin were noticed, involving secondary structure elements, binding residues and intra- as well as inter peptide residue similarities. Using surface plasmon resonance, we examined insulin binding to truncated, soluble insulin receptor A and IGF-1 receptor, but C-peptide effects on these bindings were not detectable. Two forms of the insulin receptor, differing in activation of gene transcription with regards to (pre)proinsulin and glucokinase, respectively, were also uninfluenced by C peptide. We conclude that the pattern similarities, if functional, reflect C peptide interactions with molecules other than both insulin A and B receptors and IGF-1 receptors. Any such effects are of interest in relation to reported binding interactions between insulin and C-peptide. PMID- 17115118 TI - Structural characterization of two papaya chitinases, a family GH19 of glycosyl hydrolases. AB - Two chitinases, able to use tetra-N-acetylglucosamine, chitin and chitosan as substrates, were characterized after purification from Carica papaya latex. The complete amino acid sequence of the major form and about 40% of the minor one were determined through proteolytic digestions and mass spectroscopy analysis. Sequencing demonstrated that both papaya chitinases are members of the family 19 of glycosyl hydrolases (GH19). Based on the known 3-D structures of other members of family GH19, it was expected that papaya chitinases would adopt all-alpha structures. However, circular dichroism and infrared spectroscopy indicated, for the papaya chitinases, a content of 15-20% of extended structures besides the expected 40% of alpha helices. Since the fully sequenced papaya chitinase contains a large number of proline residues the possibility that papaya chitinase contains polyproline II stretches was examined in the context of their resistance against proteolytic degradation. PMID- 17115119 TI - Claudins: multifunctional players in epithelial tight junctions and their role in cancer. AB - The molecular architecture of tight junctions has been a subject of extensive studies that have shown tight junctions to be composed of many peripheral and integral membrane proteins. Claudins have been considered the main tight junction forming proteins; however, the role they play in a series of pathophysiological events, including human carcinoma development, is only now beginning to be understood. Increasing evidence from in vitro and in vivo studies have identified the influence of claudins on tight junction structure and function, although claudins also participate in cellular contexts other than tight junctions. The aim of this review is to summarize and discuss the conceptual framework concerning claudins, focusing on the involvement of these proteins in epithelial cell polarity establishment, paracellular transport control, signal transduction and tumorigenesis. PMID- 17115120 TI - The semenogelins: proteins with functions beyond reproduction? AB - The coagulum proteins of human semen, semenogelins I and II, are secreted in abundance by the seminal vesicles. Their function in reproduction is poorly understood as they are rapidly degraded in ejaculated semen. However, more recent results indicate that it is time to put the semenogelins in a broader physiological perspective that goes beyond reproduction and fertility. PMID- 17115121 TI - Pelizaeus-Merzbacher disease: Genetic and cellular pathogenesis. AB - Pelizaeus-Merzbacher disease (PMD) and the allelic spastic paraplegia type 2 (SPG2) arise from mutations in the X-linked gene encoding myelin proteolipid protein (PLP). Analysis of mutations affecting PLP, the major protein in central nervous system myelin, has revealed previously unsuspected roles for myelinating glia in maintaining the integrity of the nervous system. The disease spectrum for PMD and SPG2 is extraordinarily broad and can be best understood by accounting not only for the wide range of mutations that can occur but also for the effects of PLP1 mutations on both cell autonomous and non-cell autonomous processes in myelinating cells. Appreciating the wide range of genetic and cellular effects of PLP1 mutations is important for patient and family counseling, understanding disease pathogenesis, and, ultimately, for developing future disease-specific therapies. PMID- 17115122 TI - Mechanical response and conformational changes of alpha-actinin domains during unfolding: a molecular dynamics study. AB - Alpha-actinin is a cytoskeleton-binding protein involved in the assembly and regulation of the actin filaments. In this work molecular dynamics method was applied to investigate the mechanical behaviour of the human skeletal muscle alpha-actinin. Five configurations were unfolded at an elongation speed of 0.1 nm/ps in order to investigate the conformational changes occurring during the extension process. Moreover, a sensitivity analysis at different velocities was performed for one of the R2-R3 spectrin-like repeat configuration extracted in order to evaluate the effect of the pulling speed on the mechanical behaviour of the molecule. Two different behaviours were recognized with respect to the pulling speed. In particular, at speed higher than 0.025 nm/ps a continuous rearrangement without evident force peaks was obtained, on the contrary at lower speed evident peaks in the range 500-750 pN were detected. R3 repeat resulted more stable than R2 during mechanical unfolding, due to the lower hydrophobic surface available to the solvent. The characterization of the R2-R3 units can be useful for the development of cytoskeleton network models based on stiffness values obtained by analyses performed at the molecular level. PMID- 17115123 TI - B1 field-insensitive transformers for RF-safe transmission lines. AB - OBJECTIVE: Integration of transformers into transmission lines suppresses radiofrequency (RF)-induced heating. New figure-of-eight-shaped transformer coils are compared to conventional loop transformer coils to assess their signal transmission properties and safety profile. MATERIALS AND METHODS: The transmission properties of figure-of-eight-shaped transformers were measured and compared to transformers with loop coils. Experiments to quantify the effect of decoupling from the B1 field of the MR system were conducted. Temperature measurements were performed to demonstrate the effective reduction of RF-induced heating. The transformers were investigated during active tracking experiments. RESULTS: Coupling to the B1 field was reduced by 18 dB over conventional loop shaped transformer coils. MR images showed a significantly reduced artifact for the figure-of-eight- shaped coils generated by local flip-angle amplification. Comparable transmission properties were seen for both transformer types. Temperature measurements showed a maximal temperature increase of 30 K/3.5 K for an unsegmented/segmented cable. With a segmented transmission line a robotic assistance system could be successfully localized using active tracking. CONCLUSION: The figure-of-eight-shaped transformer design reduces both RF field coupling with the MR system and artifact sizes. Anatomical structure close to the figure-of-eight-shaped transformer may be less obscured as with loop-shaped transformers if these transformers are integrated into e.g. intravascular catheters. PMID- 17115124 TI - Accurate quantification methods to evaluate cervical cord atrophy in multiple sclerosis patients. AB - OBJECT: Automatic accurate measurement techniques are needed to increase reproducibility in the quantification of cervical cord area (CCA) with magnetic resonance (MR) imaging in the assessment of central nervous system (CNS) atrophy in multiple sclerosis (MS) patients. MATERIALS AND METHODS: Two segmentation methods were implemented: (1) spatial mean brightness level estimation (SMBLE), and (2) partial-volume modeling (PVM). These were evaluated with the inclusion of spinal cord inclination and/or partial-volume-effect corrections. An averaged manually segmented set was considered as reference. Thirty MR studies were used to compare the different methods. A set of 15 MS patients and 15 control subjects within a two-year longitudinal study were used to evaluate cord atrophy with the best method. Statistical evaluation was made by using an intraclass correlation coefficient and Bland-Altman comparisons. RESULTS: Partial-volume modeling with spinal cord inclination correction and partial-volume spinal-cord contour contribution estimation was the most accurate method. The longitudinal test showed a 4% decrease in CCA in MS patients with no significant reduction in control subjects. CONCLUSION: The automatic PVM cord-segmentation approach, taking into consideration the spinal-cord inclination and partial-volume treatment, provides reproducibility and increased accuracy in the evaluation of cord atrophy, allowing the monitoring of changes in MS patients. PMID- 17115125 TI - Modification of gene expression of the small airway epithelium in response to cigarette smoking. AB - The earliest morphologic evidence of changes in the airways associated with chronic cigarette smoking is in the small airways. To help understand how smoking modifies small airway structure and function, we developed a strategy using fiberoptic bronchoscopy and brushing to sample the human small airway (10th-12th order) bronchial epithelium to assess gene expression (Affymetrix HG-U133A and HG 133 Plus 2.0 array) in phenotypically normal smokers (n = 16, 25 +/- 7 pack years) compared to matched nonsmokers (n = 17). Compared to samples from large (second to third order) bronchi, the small airway samples had a higher proportion of ciliated cells, but less basal, undifferentiated, and secretory cells, and contained Clara cells. Even though the smokers were phenotypically normal, microarray analysis of gene expression of the small airway epithelium of the smokers compared to the nonsmokers demonstrated up- and downregulation of genes in multiple categories relevant to the pathogenesis of chronic obstructive lung disease (COPD), including genes coding for cytokines/innate immunity, apoptosis, mucin, response to oxidants and xenobiotics, and general cellular processes. In the context that COPD starts in the small airways, these gene expression changes in the small airway epithelium in phenotypically normal smokers are candidates for the development of therapeutic strategies to prevent the onset of COPD. PMID- 17115126 TI - A high-density consensus map of barley to compare the distribution of QTLs for partial resistance to Puccinia hordei and of defence gene homologues. AB - A consensus map of barley was constructed based on three reference doubled haploid (DH) populations and three recombinant inbred line (RIL) populations. Several sets of microsatellites were used as bridge markers in the integration of those populations previously genotyped with RFLP or with AFLP markers. Another set of 61 genic microsatellites was mapped for the first time using a newly developed fluorescent labelling strategy, referred to as A/T labelling. The final map contains 3,258 markers spanning 1,081 centiMorgans (cM) with an average distance between two adjacent loci of 0.33 cM. This is the highest density of markers reported for a barley genetic map to date. The consensus map was divided into 210 BINs of about 5 cM each in which were placed 19 quantitative trait loci (QTL) contributing to the partial resistance to barley leaf rust (Puccinia hordei Otth) in five of the integrated populations. Each parental barley combination segregated for different sets of QTLs, with only few QTLs shared by any pair of cultivars. Defence gene homologues (DGH) were identified by tBlastx homology to known genes involved in the defence of plants against microbial pathogens. Sixty three DGHs were located into the 210 BINs in order to identify candidate genes responsible for the QTL effects. Eight BINs were co-occupied by a QTL and DGH(s). The positional candidates identified are receptor-like kinase, WIR1 homologues and several defence response genes like peroxidases, superoxide dismutase and thaumatin. PMID- 17115127 TI - Identification, characterization and utilization of EST-derived genic microsatellite markers for genome analyses of coffee and related species. AB - Genic microsatellites or EST-SSRs derived from expressed sequence tags (ESTs) are desired because these are inexpensive to develop, represent transcribed genes, and often a putative function can be assigned to them. In this study we investigated 2,553 coffee ESTs (461 from the public domain and 2,092 in-house generated ESTs) for identification and development of genic microsatellite markers. Of these, 2,458 ESTs (all >100 bp in size) were searched for SSRs using MISA--search module followed by stackPACK clustering that revealed a total of 425 microsatellites in 331 (13.5%) non-redundant ESTs/consensus sequences suggesting an approximate frequency of 1 SSR/2.16 kb of the analysed coffee transcriptome. Identified microsatellites mainly comprised of di-/tri-nucleotide repeats, of which repeat motifs AG and AAG were the most abundant. A total of 224 primer pairs could be designed from the non-redundant SSR-positive ESTs (excluding those with only mononucleotide repeats) for possible use as potential genic markers. Of this set, a total of 24 (10%) primer pairs were tested and 18 could be validated as usable markers. Sixteen of these markers revealed moderate to high polymorphism information content (PIC) across 23 genotypes of C. arabica and C. canephora, while 2 markers were found to be monomorphic. All the markers also showed robust cross-species amplifications across 14 Coffea and 4 Psilanthus species. The apparent broad cross-species/genera transferability was further confirmed by cloning and sequencing of the amplified alleles. Thus, the study provides an insight about the frequency and distribution of SSRs in coffee transcriptome, and also demonstrates the successful development of genic-SSRs. It is expected that the potential markers described here would add to the repertoire of DNA markers needed for genetic studies in cultivated coffee and also related taxa that constitute the important secondary genepool for coffee improvement. PMID- 17115128 TI - A microsatellite marker based linkage map of tobacco. AB - We report the first linkage map of tobacco (Nicotiana tabacum L.) generated through microsatellite markers. The microsatellite markers were predominantly derived from genomic sequences of the Tobacco Genome Initiative (TGI) through bioinformatics screening for microsatellite motives. A total of 684 primer pairs were screened for functionality in a panel of 16 tobacco lines. Of those, 637 primer pairs were functional. Potential parents for mapping populations were evaluated for their polymorphism level through genetic similarity analysis. The similarity analysis revealed that the known groups of tobacco varieties (Burley, Flue-cured, Oriental and Dark) form distinct clusters. A mapping population, based on a cross between varieties Hicks Broad Leaf and Red Russian, and consisting of 186 F2 individuals, was selected for mapping. A total of 282 functional microsatellite markers were polymorphic in this population and 293 loci could be mapped together with the morphological trait flower color. Twenty four tentative linkage groups spanning 1,920 cM could be identified. This map will provide the basis for the genetic mapping of traits in tobacco and for further analyses of the tobacco genome. PMID- 17115129 TI - Identification and mapping of a tiller inhibition gene (tin3) in wheat. AB - Tillering is one of the most important agronomic traits in cereal crops because tiller number per plant determines the number of spikes or panicles per plant, a key component of grain yield and/or biomass. In order to characterize the underlying genetic variation for tillering, we have isolated mutants that are compromised in tillering ability using ethyl methanesulphonate (EMS)-based mutagenesis in diploid wheat (Triticum monococcum subsp. monococcum). The tillering mutant, tiller inhibition (tin3) produces only one main culm compared to the wild type with many tillers. The monoculm phenotype of tin3 is due to a single recessive mutation. Genetic and molecular mapping in an F(2) population of diploid wheat located the tin3 gene on the long arm of chromosome 3A(m). One codominant RFLP marker Xpsr1205 cosegregated with tin3 in the F(2) population. Physical mapping of PSR1205 in a set of Chinese Spring deletion lines of group-3 chromosomes placed the tin3 gene in the distal 10% of the long arm of chromosome 3A, which is a recombination-rich region in wheat. The implications of the mapping of tin3 on chromosome arm 3A(m)L are discussed with respect to putative orthologs of tin3 in the 3L colinear regions across various cereal genomes and other tillering traits in grasses. PMID- 17115130 TI - Genetic mapping of the bean golden yellow mosaic geminivirus resistance gene bgm 1 and linkage with potyvirus resistance in common bean (Phaseolus vulgaris L.). AB - Bean golden yellow mosaic virus (BGYMV) is a whitefly-transmitted geminivirus of the Begomovirus family that causes important yield losses to common beans grown in tropical and sub-tropical countries of Latin America and the Caribbean. A major resistance gene that has been widely deployed in this region is the recessive locus bgm-1 that prevents the development of severe yellowing typical of the disease. In this study, we developed a co-dominant sequence-characterized amplified region (SCAR) marker, SR2, based on a previously identified random amplified polymorphic DNA (RAPD) marker that is tightly linked to the bgm-1 resistance gene and identified the position of the locus in the common bean genome through comparative mapping using two genetic maps for the species. The SR2 marker was mapped relative to bgm-1 in a segregating population of recombinant inbred lines developed from the resistant x susceptible cross of DOR476 x SEL1309. Polymorphism was shown to be based on a 37 bp insertion event in the SR2 allele associated with susceptibility compared to the allele associated with resistance and the marker mapped at a distance of 7.8 cM from the resistance gene. The SR2 marker was significantly associated with overall disease symptoms and with three of the four symptoms associated with the disease (yellowing or chlorosis, flower abortion, foliar deformation) in a greenhouse trial in Colombia with the mechanically transmissible BGYMV-Guatemala strain. In both the DOR364 x G19833 and BAT93 x Jalo EEP558 mapping populations, SR2 was located near the end of linkage group b03 (chromosome 5) suggesting a sub telomeric position. The position of the bgm-1 resistance gene was estimated to be close to that of bc-1, a strain-specific resistance gene for Bean common mosaic virus (BCMV), based on linkage of SR2 with the SCAR marker SBD5 in the DOR364 x G19833 mapping population. The implications of linkage between these two recessive resistance genes are discussed, as this is the first association between resistance genes against both a begomovirus and a potyvirus. PMID- 17115131 TI - Persistently low plasma thioredoxin is associated with meningococcal septic shock in children. AB - OBJECTIVE: To compare plasma levels of thioredoxin (Trx), TNF-alpha and IL-1 beta in children during the acute phase of meningococcal septic shock (MSS) and in convalescence. DESIGN AND SETTING: Retrospective, observational study in the paediatric intensive care unit of a postgraduate teaching hospital. PATIENTS: Thirty-five children requiring intensive care for meningococcal sepsis; paired convalescent samples from 30 survivors (median interval between samples 62 days); 25 healthy control children. MEASUREMENTS AND RESULTS: Plasma Trx levels were significantly lower in the children with MSS, both during the acute illness (5.5 ng/ml, IQR 1.4-11.4) and in convalescence (2.5 ng/ml, IQR 0.4-6.9) than controls (18.8 ng/ml, IQR 7.9-25.0). Levels of IL-1 beta and TNF-alpha were higher in patients with acute MSS (30.3 pg/ml, IQR 3.6-63.6, and 145.9 pg/ml, IQR 31.8 278.1 respectively) than controls (3.7 pg/ml, IQR 0-36.9, and 23.8 pg/ml, IQR 0 124.3, respectively). Levels fell in convalescence (3.7 pg/ml, IQR 0-25.5, 3.7 pg/ml, IQR 0-304.8, respectively). Plasma Trx was higher in non-survivors, albeit a small group (n=5), than in survivors (n=30). Trx, IL-1 beta, and TNF-alpha levels were not correlated with predicted mortality as assessed by the paediatric risk of mortality (PRISM) score. CONCLUSIONS: Children with MSS exhibit persistently low plasma levels of Trx during acute illness and in convalescence. PMID- 17115132 TI - Intestinal motility disturbances in intensive care patients pathogenesis and clinical impact. AB - BACKGROUND: Gastrointestinal motility disturbances in critically ill patients are frequent in the ICU setting, causing considerable discomfort and are associated with increased rates of morbidity and mortality. This review focuses on the pathophysiological basis of intestinal motility, the major patterns of pathological motility alterations, the impact on patient outcome, and current therapeutic options. DISCUSSION: Intestinal motility is controlled by the enteric nervous system, modulated by hormones and extrinsic afferent and efferent neurons. Pathological motility disturbances can affect the stomach, small bowel, and colon separately or in combination. Changes in esophageal motor activity contribute to the aspiration of gastric juice, whereas early enteral feeding most frequently fails due to gastric intolerance. Disturbances in digestive and interdigestive motility patterns and the inability to switch motor activity from the interdigestive to the digestive pattern also contribute to feeding disability and thus to increased morbidity and mortality as well. CONCLUSIONS: The therapeutic options for motility disturbances in critically ill patients include the adjustment of electrolyte imbalances, tailored fluid management, early enteral feeding, appropriate management of catecholamines and drugs used for analgosedation, and prokinetic drugs. Unfortunately, the therapeutic options for treating motility disturbances in ICU patients are still limited. This situation requires careful assessment of ICU patients with respect to gut motility disturbances and their pathophysiological mechanisms and an individually tailored treatment to prevent further aggravation of existing motility disturbances. PMID- 17115133 TI - Danger of helmet continuous positive airway pressure during failure of fresh gas source supply. AB - OBJECTIVE: To assess the behavior of different helmets after discontinuation of fresh gas flow by disconnection at the helmet inlet, flow generator, or gas source. DESIGN AND SETTING: Randomized physiological study in a university research laboratory. PATIENTS: Five healthy volunteers. INTERVENTION: CPAP (FIO2 50%, PEEP 5 cmH2O) delivered in random sequence with three different helmets: 4Vent (Rusch), PN500 (Harol), CaStar (StarMed) with antisuffocation valve open or locked. For each helmet all three disconnections were randomly employed up to 4 min. MEASUREMENTS AND RESULTS: During flow disconnection we measured: respiratory rate and tidal volume by respitrace; inspiratory and expiratory CO2 concentration, and FIO2 from a nostril; SpO2 by pulse oxymetry. Independently of the site of disconnection we observed a fast increase in CO2 rebreathing and minute ventilation, associated with a decrease in inspired O2 concentration. In the absence of an operational safety valve, larger helmet size and lower resistance of the inlet hose resulted in slower increase in CO2 rebreathing. The presence of the safety valve limited the rebreathing of CO2, and the increase in minute ventilation but did not protect from a decrease in FIO2 and loss of PEEP. CONCLUSIONS: While the use of a safety valve proved effective in limiting CO2 rebreathing, it did not protect from the risk of hypoxia related to decrease in FIO2 and loss of PEEP. In addition to a safety antisuffocation valve, a dedicated monitoring and alarming systems are needed to employ helmet CPAP safely. PMID- 17115134 TI - Intensive care course after stage 1 Norwood procedure: are there early predictors of failure? AB - OBJECTIVE: The purpose of this study was to review the early postoperative course of stage 1 Norwood with Blalock-Taussig shunt (BTS) or right ventricle-to pulmonary artery conduit (RVPA) and to identify early predictors of failure. MATERIAL AND METHODS: A retrospective analysis was conducted in 33 consecutive neonates who underwent BTS (n=19) or RVPA (n=14) stage 1 Norwood procedure between 2000 and 2005. Pre-, peri-, and postoperative data included: hourly hemodynamics and blood gases, pulmonary to systemic flow ratio, duration of mechanical ventilatory and inotrope support, intensive care and hospital stay. Failure was defined as death or transplantation. RESULTS: Thirteen patients failed the procedure (39.4%): 10 BTS (52.6%) and 3 RVPA (21.4%). Failure decreased from 61.1% in 2000-2002 to 13.3% in 2003-2005 and was associated with: low systolic, mean and diastolic blood pressure, urine output, pH, base excess, bicarbonates, and high pulmonary to systemic flow ratio within 24 h postoperatively. Arterial oxygen and CO2 pressure, and oxygen saturation did not differ with failure. RVPA had higher diastolic blood pressure and more stable hemodynamics despite similar pulmonary to systemic flow ratio. Duration of mechanical ventilation, inotrope support, intensive care stay were shorter in RVPA. Postoperative echographic ventricular dysfunction and tricuspid regurgitation grade were correlated with failure. CONCLUSIONS: Excessive pulmonary to systemic flow ratio and low blood pressure are associated with failure. High diastolic blood pressure more than low pulmonary to systemic flow ratio seems to account for more favorable outcomes in RVPA compared to BTS procedure. PMID- 17115135 TI - Impact of antifungal treatment on Candida-Pseudomonas interaction: a preliminary retrospective case-control study. AB - OBJECTIVE: A pathogenic interaction between Candida albicans and Pseudomonas aeruginosa has recently been demonstrated. In addition, experimental and clinical studies identified Candida spp. tracheobronchial colonization as a risk factor for P. aeruginosa pneumonia. The aim of this study was to determine the impact of antifungal treatment on ventilator-associated pneumonia (VAP) or tracheobronchial colonization due to P. aeruginosa. DESIGN AND SETTING: Retrospective observational case-control study conducted in a 30-bed ICU during a 1-year period. PATIENTS AND METHODS: One hundred and two patients intubated and ventilated for longer than 48 h with tracheobronchial colonization by Candida spp. Routine screening for Candida spp. and P. aeruginosa was performed at ICU admission and weekly. Antifungal treatment was based on medical staff decisions. Patients with P. aeruginosa VAP or tracheobronchial colonization were matched (1:2) with patients without P. aeruginosa VAP or tracheobronchial colonization. In case and control patients, risk factors for P. aeruginosa VAP or tracheobronchial colonization were determined using univariate and multivariate analyses. RESULTS: Thirty-six patients (35%) received antifungal treatment. Nineteen patients (18%) developed a P. aeruginosa VAP or tracheobronchial colonization, and all were successfully matched. Antifungal treatment [31% vs 60%; p=0.037, OR (95% CI)=0.67 (0.45-0.90)], and duration of antifungal treatment (7+/-11 vs 14+/-14 days; p=0.045, in case and control patients respectively) were significantly associated with reduced risk for P. aeruginosa VAP or tracheobronchial colonization. Antifungal treatment was the only variable independently associated with P. aeruginosa VAP or tracheobronchial colonization (OR=0.68, 95% CI=0.49-0.90, p=0.046). CONCLUSION: In patients with Candida spp. tracheobronchial colonization, antifungal treatment may be associated with reduced risk for P. aeruginosa VAP or tracheobronchial colonization. PMID- 17115136 TI - Ligands selective for alpha4beta2 but not alpha3beta4 or alpha7 nicotinic receptors generalise to the nicotine discriminative stimulus in the rat. AB - RATIONALE: Nicotine produces behavioural effects that are potentially related to its interaction with diverse nicotinic acetylcholine receptor populations. Evidence from gene deletion studies suggests that the interoceptive stimulus properties of nicotine are mediated by heteromeric high-affinity receptors containing alpha4beta2 subunits. Mice lacking beta2 subunits do not discriminate nicotine (Shoaib et al., Neuropharmacology, 42:530-539, 2002), and nicotine does not elicit dopamine release in these animals (Grady et al., J Neurochem, 76:258 268, 2001). The stimulus properties of nicotine can be detected in rats using a two-lever operant drug discrimination paradigm, allowing them to be classified pharmacologically using ligands with selectivity for receptors containing alpha4beta2, alpha3beta4 or alpha7 subunits. MATERIALS AND METHODS: Rats trained to discriminate 0.4 mg/kg nicotine from vehicle were given the nicotinic receptor agonists, cytisine, varenicline, TC2559, ABT-594, A-85380 (all having high affinity but varying selectivity for alpha4beta2-containing receptors), and WO 03/062224 and WO 01/60821A1 (selective for beta4- and alpha7-containing receptors, respectively). In separate studies, WO 03/062224 was used as the training stimulus. RESULTS: Nicotine, TC-2559, A-85380 and ABT-594 showed dose dependent and complete stimulus substitution, whilst WO 03/062224 and WO 01/60821A1 were completely without effect. Cytisine and varenicline showed partial generalisation, consistent with their partial agonist activity at nicotinic receptors eliciting dopamine release in rat striatal slices. After almost 50 training sessions with WO 03/062224, there was no clear evidence that an alpha3beta4 receptor agonist could sustain a discriminable stimulus. CONCLUSION: Substitution to the nicotine discriminative stimulus required high affinity and high intrinsic activity at beta2 but not at beta4- or at alpha7 containing nicotinic receptors. PMID- 17115137 TI - mRNA transport to and translation in neuronal dendrites. AB - Transport of mRNA is an important biological process in all cells that sets up gradients of translated protein from the site of mRNA docking and translation. Neurons are highly polarized cells where the targeted movement of RNAs and local translation at that site have been shown to be integral to the proper functioning of the neuron. Indeed, this specialized biological function for localized RNAs in particular neurons may in part confer a selective advantage on these cells such that they "out-compete" others in the race to establish synaptic connectivity. In this mini-review we highlight some of the salient features of RNA targeting and translation in neurons. PMID- 17115138 TI - Development of a sensitive micro-magnetic chemiluminescence enzyme immunoassay for the determination of carcinoembryonic antigen. AB - A micro-magnetic chemiluminescence (CL) enzyme immunoassay with high sensitivity, selectivity, and reproducibility was developed for the determination of the tumor marker, carcinoembryonic antigen (CEA) in human serum. A sandwich scheme assay has been utilized with fluorescein isothiocyanate antibody (FITC)-labeled anti CEA antibody and alkaline phosphate (ALP)-labeled anti-CEA antibody being used in the CL detection. The CL signal produced by the emission of photons from 4 methoxy-4-(3-phosphate-phenyl)-spiro-(1,2-dioxetane-3,2'-adamantane) (AMPPD) was directly proportional to the amount of analyte present in a sample solution. The influences of the reaction time of antigen with antibody, the reaction time of substrate with label, the dilution ratio of ALP-labeled anti-CEA antibody, the concentration of FITC-labeled anti-CEA antibody, and other relevant variables upon the CL signal were examined and optimized. The CL responses depended linearly on the CEA concentration over the range from 2 to 162 ng mL-1 in a logarithmic plot. Assay sensitivity as low as 0.69 ng mL-1 was achieved. A coefficient of variance of less than 13% was obtained for intra- and inter-assay precision. This method has been successfully applied to the analysis of CEA in human serum. According to the procedure based on spiked standards, the recoveries obtained were 80-110%. Comparison experiments were carried out with the commercially available CEA chemiluminescence immunoassay. Satisfactory results were obtained according to a paired t-test method (t value80%) and effluent concentrations of, e.g., diclofenac, ketoprofen, ranitidine, gemfibrozil, bezafibrate, pravastatin, and ofloxacin were steadier than for the conventional system. Occasionally removal efficiency was very similar, and high, for both treatments (e.g. for ibuprofen, naproxen, acetaminophen, paroxetine, and hydrochlorothiazide). The antiepileptic drug carbamazepine was the most persistent pharmaceutical and it passed through both the MBR and CAS systems untransformed. Because there was no washout of biomass from the reactor, high-quality effluent in terms of chemical oxygen demand (COD), ammonium content (N-NH(4)), total suspended solids (TSS), and total organic carbon (TOC) was obtained. PMID- 17115142 TI - The curse of having a bad supervisor. PMID- 17115141 TI - A new sample substrate for imaging and correlating organic and trace metal composition in biological cells and tissues. AB - Many disease processes involve alterations in the chemical makeup of tissue. Synchrotron-based infrared (IR) and X-ray fluorescence (XRF) microscopes are becoming increasingly popular tools for imaging the organic and trace metal compositions of biological materials, respectively, without the need for extrinsic labels or stains. Fourier transform infrared microspectroscopy (FTIRM) provides chemical information on the organic components of a material at a diffraction-limited spatial resolution of 2-10 microm in the mid-infrared region. The synchrotron X-ray fluorescence (SXRF) microprobe is a complementary technique used to probe trace element content in the same systems with a similar spatial resolution. However to be most beneficial, it is important to combine the results from both imaging techniques on a single sample, which requires precise overlap of the IR and X-ray images. In this work, we have developed a sample substrate containing a gold grid pattern on its surface, which can be imaged with both the IR and X-ray microscopes. The substrate consists of a low trace element glass slide that has a gold grid patterned on its surface, where the major and minor parts of the grid contain 25 and 12 nm gold, respectively. This grid pattern can be imaged with the IR microscope because the reflectivity of gold differs as a function of thickness. The pattern can also be imaged with the SXRF microprobe because the Au fluorescence intensity changes with gold thickness. The tissue sample is placed on top of the patterned substrate. The grid pattern's IR reflectivity image and the gold SXRF image are used as fiducial markers for spatially overlapping the IR and SXRF images from the tissue. Results show that IR and X-ray images can be correlated precisely, with a spatial resolution of less than one pixel (i.e., 2-3 microns). The development of this new tool will be presented along with applications to paraffin-embedded metalloprotein crystals, Alzheimer's disease, and hair composition. PMID- 17115143 TI - Determination of major, minor and trace elements in cobalt-substituted lithium nickelate ceramic powders by inductively coupled plasma optical emission spectrometry. AB - An analytical method was developed for the determination of three major (Li, Ni and Co) and fourteen minor or trace elements (Al, Ba, Ca, Cu, Cr, Fe, K, Mg, Mn, Na, Si, Sr, Ti and V) in LiNi1-xCoxO2 (x=0.2-0.8) ceramic powders by inductively coupled plasma optical emission spectrometry. Sample dissolution was achieved by 25% nitric acid digestion in a microwave oven. For each element, an analytical line free from spectral interferences was selected. A detailed study of matrix effects over a wide interval of total excitation energy (TEE) lines (1.62-16.50 eV) was performed at near-robust plasma conditions. A remarkable enhancement in atomic lines with TEE<4 eV was noticed, whereas a significant reduction in atomic and ionic lines with TEE>4 eV was observed. The extrapolation to infinite dilution method was successfully used to overcome these nonspectroscopic interferences. Detection limits (3sigma) varied from 0.21 mg kg-1 for Sr to 49.7 mg kg-1 for Na. The precision of determination (obtained as the relative standard deviation) was lower than 1% for the major elements Li, Ni and Co and between 0.69 and 10% for minor and trace elements. The accuracy of the method ranged from 91 to 101% for major elements, and from 90 to 110%, or close to this range, for most of the impurities in both of the samples studied. PMID- 17115144 TI - Selective detection of dopamine in the presence of ascorbic acid by use of glassy carbon electrodes modified with both polyaniline film and multi-walled carbon nanotubes with incorporated beta-cyclodextrin. AB - A simple, sensitive, and reliable method based on a combination of multi-walled carbon nanotubes with incorporated beta-cyclodextrin (beta-CD-MWNTs) and a polyaniline (PANI) film-modified glassy-carbon (GC) electrode has been successfully developed for determination of dopamine (DA) in the presence of ascorbic acid (AA). The PANI film had good anti-interference properties and long term stability, because of the permselective and protective properties of the conducting redox polymer film. The acid-treated MWNTs with carboxylic acid functional groups promoted the electron-transfer reaction of DA and inhibited the voltammetric response of AA. Sensitive detection of DA was further improved by the preconcentration effect of formation of a supramolecular complex between beta CD and DA. The analytical response of the beta-CD-MWNTs/PANI film to the electrochemical behavior of DA was, therefore, better than that of a MWNTs/PANI film, a PANI film, or a bare glassy-carbon (GC) electrode. Under the conditions chosen a linear calibration plot was obtained in the range 1.0 x 10(-7)-1.0 x 10( 3) mol L(-1) and the detection limit was 1.2 x 10(-8) mol L(-1). Interference from AA was effectively eliminated and the sensitivity, selectivity, stability, and reproducibility of the electrodes was excellent for determination of DA. PMID- 17115145 TI - Method optimization for determination of selected perfluorinated alkylated substances in water samples. AB - In recent years perfluorinated alkylated substances (PFAS) have appeared as a new class of global pollutant. Besides being an industrially important group of compounds, PFAS are regarded as highly toxic and extraordinarily persistent chemicals that pervasively contaminate human blood and wildlife throughout the world. They are therefore regarded as PBT (persistent, bioaccumulative, and toxic) chemicals. Two comprehensive methods have been developed for determination of eleven of the most environmentally relevant PFAS (seven perfluoroalkylcarboxylates, two perfluoroalkylsulfonates, and two perfluoroctanesulfonamides) in aqueous samples. The compounds were isolated by liquid-liquid extraction (LLE) and solid-phase extraction (SPE), and identification and quantification of the target analytes were achieved by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS). With LLE detection limits ranged from 0.26 to 0.62 ng L(-1) for enrichment of 900 mL water samples; recovery of PFAS with a carbon chain longer than C7 was excellent (80-93%). With SPE, carboxylates with carbon chains A, 98T > C, 766G > A) and UGT2B7 (211G > T, 802C > T, 1192G > A) SNPs were determined in WA (n = 133, 5 countries), PNG (n = 153), and NA (n = 350, 4 ethnic groups) individuals. RESULTS: The UGT1A9 variant alleles were not common in the study populations. None of the SNPs were present in WA and PNG. Among NA, all 3 SNPs were present (1% each) in Asian-Americans, while 98T > C was present only in Caucasian-Americans (1%) and Hispanic-Americans (1%). Regarding UGT2B7 SNPs, the prevalence of 802C > T was 21% in WA, 28% in PNG, and 28-52% in NA. The SNP 211G > T was present only in Asian-Americans (9%) and Hispanic-Americans (2%), while 1192G > A was not present in any of the subjects. No significant linkage was observed at UGT1A9, UGT2B7, and between both the loci in any of the study populations. CONCLUSIONS: Taken together, the UGT1A9 UGT2B7 polymorphism profile in WA and PNG populations is similar to African Americans, but different from Asian-Americans. It is important to determine if these differences, along with previously reported differences in cytochrome P450 2B6 allele frequencies, are associated with altered metabolism/effectiveness of artemisinin drugs. PMID- 17115151 TI - Changes induced by hyperosmotic mannitol in cerebral endothelial cells: an atomic force microscopic study. AB - Understanding the reaction of living cells in response to different extracellular stimuli, such as hyperosmotic stress, is of primordial importance. Mannitol, a cell-impermeable non-toxic alcohol, has been used successfully for reversible opening of the blood-brain barrier in hyperosmotic concentrations. In this study we analyzed the effect of hyperosmotic mannitol on the shape and surface structure of living cerebral endothelial cells by atomic force microscope imaging technique. Addition of clinically relevant concentrations of mannitol to the culture medium of the confluent cells induced a decrease of about 40% in the observed height of the cells. This change was consistent both at the nuclear and peripheral region of the cells. After mannitol treatment even a close examination of the contact surface between the cells did not reveal gap between them. We could observe the appearance of surface protrusions of about 100 nm. By force measurements the elasticity of the cells were estimated. While the Young's modulus of the control cells appeared to be 8.04 +/- 0.12 kPa, for the mannitol treated cells it decreased to an estimated value of 0.93 +/- 0.04 kPa which points to large structural changes inside the cell. PMID- 17115152 TI - Dimerization of Neu/Erb2 transmembrane domain is controlled by membrane curvature. AB - Secondary structures of the proto-oncogenic Neu/ErbB2 transmembrane segment and its mutant analogue have been determined in phospholipids. It is found that the mutated peptide possesses less helical character possibly due to the valine/glutamic acid point mutation. Embedding peptides in lipid systems whose topology can change from small (100-200 A) tumbling objects to bilayer discs of 450 A diameter leads to the finding that coiled-coil interactions are only observed in the presence of a bilayer membrane of low curvature, independent of mutation. This strongly suggests that any event that may change membrane topology can therefore perturb the dimerization/ologomerization and subsequent phosphorylation cascade leading to cell growth or cancer processes. PMID- 17115153 TI - Evaluation of the utility of the Ponseti method of correction of clubfoot deformity in a developing nation. AB - Clubfoot is the commonest congenital deformity in babies. More than 100,000 babies are born worldwide each year with congenital clubfoot. Around 80% of the cases occur in developing nations. We treated 154 feet [mean Pirani score (total) 5.57] in 96 children (78 males, 18 females) by the Ponseti method from January 2003 to December 2005. A prospective follow-up for a mean duration of 19.5 months (range 6-32 months) was undertaken. After six months of treatment the Pirani score was reduced to zero for all patients. The results show that corrective surgery, sometimes multiple, can be avoided in most cases which are usually associated with the development of a stiff, painful foot. Low socio-economic status and illiteracy prevailing in developing nations increases the prevalence of neglected clubfoot that is still harder to correct. Integration into various programs and proper use of available resources can decrease neglected clubfoot and improve chances of successful and timely correction of deformity. Bracing constitutes an important part of treatment and proper motivation and education of the parents mitigates the chances of losing correction. The Ponseti method of correcting clubfoot is especially important in developing countries, where operative facilities are not available in the remote areas and well-trained physicians and personnel can manage the cases effectively with cast treatment only. PMID- 17115154 TI - Influence of the tibial stem design on bone density after cemented total knee arthroplasty: a prospective seven-year follow-up study. AB - We prospectively measured the changes in bone mineral density (BMD) in the proximal tibia of 20 total knee arthroplasties, ten with cruciform stems and ten with cylindrical stems. The measurements were made one, four and seven years after surgery. We observed a uniform density decrease in three regions of interest from one to seven years of follow-up. Cylindrical stems showed an asymmetrical density decrease between the three regions of interest, with no change in the central region, a slight decrease in the lateral region, and large decrease in the medial region. Multivariate analysis with general linear model showed the stem type factor as statistically significant for medial region of interest (p = 0.006). The cylindrical stem produces heterogeneous BMD changes under the tibial platform in knee arthroplasties, and this could be a potential risk factor for asymmetrical subsidence of this component. PMID- 17115155 TI - Effect of shed blood retransfusion on pulmonary perfusion after total knee arthroplasty: a prospective controlled study. AB - Postoperative shed blood retransfusion (autotransfusion) is a commonly used salvage method following major surgical operations, such as total knee arthroplasty (TKA). The systemic effects of shed blood are still unclear. We studied the effect of residual substances in the retransfused shed blood, on lung perfusion after TKA. Fifteen unilateral and one bilateral TKAs were performed with autotransfusion (the study group) and 15 unilateral and three bilateral TKAs were performed in a control group. Lung X-rays, arterial blood gases (ABG), D dimer values, and lung perfusion scintigraphies were performed preoperatively and postoperatively. A mean of 300.0 +/- 335.6 ml of bank blood was needed in the autotransfusion group and a mean of 685.7 +/- 365.5 ml of bank blood was needed in the control group (p=0.001). There was a postoperative segmental perfusion defect at the lateral segment of the superior lobe of the left lung in one patient of the control group and he also had risk factors for thrombosis. Although both groups had a decrease in lung perfusion postoperatively, there were no significant differences among the groups regarding the lung perfusion scintigraphy, chest X-rays, ABG, and D-dimer values. In conclusion, although pulmonary perfusion diminishes following TKA, shed blood retransfusion does not add any risk to pulmonary perfusion. PMID- 17115156 TI - Experiences with computer navigated total knee arthroplasty. AB - The successful outcome of total knee arthroplasty (TKA) is very much dependent on precise positioning of the components. Inaccuracy may result in complaints as well as in early mechanical failure. Between March 2003 and September 2005, 69 TKA procedures were performed by the computer navigated technique. The postoperative outcome of this cohort was compared with the same number of TKAs done by the traditional technique. The lower limb anatomical axis was determined in all cases pre- and postoperatively by weight-bearing anteroposterior (AP) and lateral full length X-rays. The positions of femoral and tibial components were recorded. Comparing the data in the navigation group on the AP view, 96.6% of femoral and 96.9% of tibial components and on the lateral view in 95.4% of femoral and in 95.4% of tibial components, the overall postoperative axis in 95.4% fell in the range considered in the literature as optimal. In the traditional group on the AP view, 75.7% of femoral and 68.1% of tibial components and on the lateral view 81.8% of femoral and 63.6% of tibial components, the overall postoperative axis in 60.6% fell between the values considered optimal in the literature. It seems to be proven that the computer navigated total knee arthroplasty technique ensures positioning of components significantly more precisely compared with the traditional surgical method. Accuracy of navigation depends on the software used, on the correct detection of anatomical reference points, and on a potentially uneven thickness of the cement layer during final insertion of the components. The computer navigated technique does not substitute professional skill and experience, since it merely transmits information for the surgeon. The decision is in the hands of the doctor during the entire procedure. The real benefits of the computer navigated technique require further research and can be determined only after long-term analyses. PMID- 17115157 TI - A first-in-man phase I tolerability and pharmacokinetic study of the cyclin dependent kinase-inhibitor AZD5438 in healthy male volunteers. AB - AZD5438 is a novel cyclin-dependent kinase inhibitor with preclinical pharmacodynamic (PD) activity against a range of human tumour xenografts. A first in-man tolerability and pharmacokinetic (PK) study involving single ascending doses of AZD5438 was conducted in healthy male volunteers. Single oral doses ranging from 5 to 160 mg were studied in 23 subjects. Dose-limiting nausea and vomiting occurred at 160 mg in the absence of prophylactic anti-emetics. The maximum tolerated dose (the dose at which no dose limiting toxicities occurred) was 80 mg, and the maximum well-tolerated dose was deemed to be 60 mg, which was associated with grade1 nausea but no vomiting. Tmax occurred between 0.5-3.0 hours with a relatively short plasma half-life of 1-3 h. The coefficient of variation of exposures within a dose level ranged from 22-71% (AUC) to 16-63% (C max), and exposure increased with increasing dose across the doses studied. <1% of the parent compound was excreted in the urine, suggesting metabolism as the major clearance mechanism. The maximum well-tolerated dose and a number of doses below this level will be taken forward into a PD study using normal tissue biomarkers in humans to determine proof of AZD5438's action on the cell cycle. The pharmacokinetic profile of AZD5438 determined within this study will be used to guide the time-points for PD analysis within the planned PD study. PMID- 17115159 TI - Evaluation of in-stent restenosis in proximal coronary arteries with multidetector computed tomography (MDCT). AB - The purpose of this study was to assess the ability of 16-slice computed tomography (CT) to detect in-stent restenosis of proximal coronary arteries. From November 2002 to April 2004, 134 consecutive patients with proximal stents (3.25 +/- 0.47 mm) were prospectively studied. Multidetector CT (MDCT) was performed 24 h (baseline) and 6 months after angioplasty and analysed by two radiologists blinded to the results of the coronary angiography. Sensitivity, specificity, positive and negative predictive values for in-stent restenosis were compared with conventional quantitative coronary angiography (QCA). Stenosis with a diameter >or=50% was considered diagnostic of in-stent restenosis. The CT analysis was performed in 131 and 114 patients at baseline and 6 months, respectively. The in-stent lumen was evaluable in 111 (121 stents) and 99 patients (108 stents) at baseline and 6 months, respectively. The prevalence of in-stent restenosis was 22.5%. Restenoses were correctly identified in 91.7 and 87.5% by the two radiologists. The sensitivity, specificity, positive and negative predictive values for the assessment of significant in-stent restenosis were 92, 67, 43, 97% and 87, 66, 41, 95% for the radiologists, respectively. MDCT is a potential non-invasive technique for the screening of in-stent restenosis of proximal coronary arteries that needs further improvements. PMID- 17115158 TI - Activating and inhibitory FcgammaRs in autoimmune disorders. AB - Autoimmune disorders are characterized by the destruction of self-tissues by the immune system. Multiple checkpoints are in place to prevent autoreactivity under normal circumstances. Coexpression of activating and inhibitory Fc receptors (FcR) represents such a checkpoint by establishing a threshold for immune cell activation. In many human autoimmune diseases, however, balanced FcR expression is disturbed. Analysis of murine model systems provides strong evidence that aberrant FcR expression can result in uncontrolled immune responses and the initiation of autoimmune disease. This review will summarize this data and explain how this information might be used to better understand human autoimmune diseases and to develop novel therapeutic strategies. PMID- 17115160 TI - Quantification of bronchial dimensions at MDCT using dedicated software. AB - This study aimed to assess the feasibility of quantification of bronchial dimensions at MDCT using dedicated software (BronCare). We evaluated the reliability of the software to segment the airways and defined criteria ensuring accurate measurements. BronCare was applied on two successive examinations in 10 mild asthmatic patients. Acquisitions were performed at pneumotachographically controlled lung volume (65% TLC), with reconstructions focused on the right lung base. Five validation criteria were imposed: (1) bronchus type: segmental and subsegmental; (2) lumen area (LA)>4 mm2; (3) bronchus length (Lg) > 7 mm; (4) confidence index - giving the percentage of the bronchus not abutted by a vessel (CI) >55% for validation of wall area (WA) and (5) a minimum of 10 contiguous cross-sectional images fulfilling the criteria. A complete segmentation procedure on both acquisitions made possible an evaluation of LA and WA in 174/223 (78%) and 171/174 (98%) of bronchi, respectively. The validation criteria were met for 56/69 (81%) and for 16/69 (23%) of segmental bronchi and for 73/102 (72%) and 58/102 (57%) of subsegmental bronchi, for LA and WA, respectively. In conclusion, BronCare is reliable to segment the airways in clinical practice. The proposed criteria seem appropriate to select bronchi candidates for measurement. PMID- 17115161 TI - Effects of injection rate and dose on image quality in time-resolved magnetic resonance angiography (MRA) by using 1.0M contrast agents. AB - In time-resolved MRA (TR MRA), injection parameters and contrast agent (CA) dose are important factors influencing image quality. In this study, three different injection schemes with different CA volumes were evaluated in 12 healthy volunteers. Injection rates between 0.2 and 0.8 ml/s were evaluated with CA volumes of 10 and 20 ml. To measure circulatory parameters, cine cardiac MRI was performed before each exam. Spatial resolution could be reduced to 2 x 1.4 x 2 mm3, temporal resolution was 2.25 s/frame. To exclude signal saturation at high CA concentrations, a phantom with fixed CA concentrations was placed in the field of view. SNR was measured, and the area under the curve of the arterial signal of the different injection schemes was calculated. Results showed the largest diagnostic window at a relatively slow injection rate of 0.4 ml/s and a CA volume of 10 ml. Circulatory parameters have an important impact on CA arrival, so delay times have to be set depending on these parameters. PMID- 17115162 TI - Energetic differences between viable and non-viable myocardium in patients with recent myocardial infarction are not an effect of differences in wall thinning- a multivoxel (31)P-MR-spectroscopy and MRI study. AB - To evaluate multivoxel (31)P-MR spectroscopy (MRS) for assessment of energy metabolism in patients with myocardial infarction (MI) in correlation to left ventricular (LV) wall thickness and the outcome of revascularization. Thirty patients with subacute anterior myocardial infarction and planned revascularization were enrolled. 3D-chemical shift imaging was applied to determine PCr/ATP ratios in two areas: infarcted/anterior and noninfarcted/septal myocardium. MRI was used to evaluate LV function and wall thickness, and was repeated 6 months after revascularization to assess myocardial viability. Fifteen volunteers were controls. Fifteen patients showed normalization of wall motion abnormalities after revascularization (Group 1; viable), 15 not (Group 2; non viable). Regarding infarcted/anterior myocardium, Group 2 had lower PCr/ATP ratios (0.81 +/- 0.60 vs 1.17 +/- 0.25), and PCr/ATP ratios were reduced in both groups compared to controls (1.45 +/- 0.29). Regarding noninfarcted/septal myocardium, again Group 2 had lower ratios (0.93 +/- 0.53 vs 1.31 +/- 0.38); however, compared to controls (1.51 +/- 0.32) a reduction of PCr/ATP ratios was only found in Group 2. For both myocardial regions, no correlations between PCr/ATP ratios and LV wall thickness were detected. The more severe energetic alteration in irreversibly damaged myocardium is not an effect of differences of wall thinning. Additional alterations of noninfarcted, adjacent myocardium can be detected. PMID- 17115163 TI - Hip pain in adults: MR imaging appearance of common causes. AB - To determine the exact origin of hip pain can be challenging. Symptoms apparently originating from the hip may arise from the pelvis, the sacroiliac joint, the lumbar spine, periarticular structures such as muscles and bursae, or from unexpected sites such as the abdominal wall, the genitourinary tract, or the retroperitoneal space. This article reviews the differential diagnosis of hip pain arising from the hip and surrounding structures and the role of different imaging methods with emphasis on magnetic resonance imaging where most recent advances have occurred. PMID- 17115164 TI - Quantitative prediction of contrast enhancement from test bolus data in cardiac MSCT. AB - The purpose of this study was to evaluate a new algorithm for the prediction of contrast enhancement from test bolus data in cardiac multislice spiral computed tomography (MSCT). An algorithm for the prediction of contrast enhancement using test bolus data was developed. A total of 30 consecutive patients (15 male, 69.5 +/- 9.6 years) underwent cardiac MSCT (12 x 0.75 mm, 120 kV, 500 mAs(eff.)) with a biphasic contrast material injection protocol. Contrast timing was derived from a standard 20 ml test bolus injection. Based on the test bolus time attenuation curves, expected enhancement values were computed for the ascending and descending aorta and the pulmonary trunk and compared with measured data from the cardiac CT scan. At the level of the test bolus measurement in the ascending aorta, the corresponding attenuation values were 309.4 +/- 49.6 Hounsfield Units (HU) for the predicted and 285.6 +/- 42.6 HU for the measured attenuation, respectively. The mean deviation between predicted and measured CT values was 32.8 +/- 48.2 HU (upper and lower limits of agreement 101.4/-53.8 HU), indicating a slight systematic tendency for overestimation. For 80% of the patients the prediction error was less than 50 HU. Prediction of contrast enhancement in cardiac MSCT from test bolus data is feasible with a relatively small mean deviation; 80% of the predictions were within a range that might be acceptable for routine clinical application. PMID- 17115165 TI - Efficacy of continuous thrombolytic therapy for portal vein occlusion after hepatobiliary pancreatic surgery. PMID- 17115166 TI - Do flat detector cardiac X-ray systems convey advantages over image-intensifier based systems? Study comparing X-ray dose and image quality. AB - The recent introduction of "flat-panel detector" (FD)-based cardiac catheterisation laboratories should offer improvements in image quality and/or dose efficiency over X-ray systems of conventional design. We compared three X ray systems, one image-intensifier (II)-based system (system A), and two FD-based designs (systems B and C), assessing their image quality and dose efficiency. Phantom measurements were performed to assess dose rates in fluoroscopy and cine acquisition. Phantom dose rates were broadly similar for all systems, with all systems classified as offering "low" dose rates in fluoroscopy on standard phantoms. Patient X-ray dose rate and subjective image quality was assessed for 90 patients. Dose area product (DAP) rates were similar for all systems, except system C, which had a lower DAP rate in fluoroscopy. In terms of subjective image quality, the order of preference was (best to worst): system C, system A, system B. This study indicates that the use of an FD detector does not infer an automatic improvement in image quality or dose efficiency over II based designs. Specification and configuration of all of the components in the X-ray system contribute to the dose levels used and image quality achieved. PMID- 17115167 TI - Placement of endovascular stent-grafts for emergency repair of acute traumatic aortic rupture: a single-centre experience. AB - The aim of this retrospective study was to evaluate mid- and long-term results of endovascular stent-graft placement for emergency repair of acute traumatic thoracic aortic rupture. From 1996 through 2005, 22 consecutive patients (mean age: 38.7 years) underwent endovascular repair of acute traumatic thoracic aortic rupture located at the aortic isthmus in most cases. All patients were at high surgical risk due to severe associated injuries. The endografts were inserted via femoral or iliac artery access under fluoroscopic guidance. Follow-up was performed postinterventionally, at 6 and 12 months and yearly thereafter, and included clinical examination and computed tomography (CT) scans. Technical and clinical success rates were 86.3%. Mean follow-up was 31.7 months. Three patients developed early type I endoleak due to the inability of the rigid graft to adapt to the curved aortic contour. In two of them conversion to open surgery was necessary. One patient had late type I endoleak and died. No other complications were observed. The outcome was successful in most patients. The mid- and long term results of our current study are promising. However, early type I endoleak represents a problem, especially in adolescent patients with a marked curvature of the aortic arch. PMID- 17115168 TI - Increased expression of phosphate-activated glutaminase in hippocampal neurons in human mesial temporal lobe epilepsy. AB - Patients with mesial temporal lobe epilepsy (MTLE) have increased basal concentrations of extracellular glutamate in the epileptogenic versus the non epileptogenic hippocampus. Such elevated glutamate levels have been proposed to underlie the initiation and maintenance of recurrent seizures, and a key question is what causes the elevation of glutamate in MTLE. Here, we explore the possibility that neurons in the hippocampal formation contain higher levels of the glutamate synthesizing enzyme phosphate-activated glutaminase (PAG) in patients with MTLE versus patients with other forms of temporal lobe epilepsy (non-MTLE). Increased PAG immunoreactivity was recorded in subpopulations of surviving neurons in the MTLE hippocampal formation, particularly in CA1 and CA3 and in the polymorphic layer of the dentate gyrus. Immunogold analysis revealed that PAG was concentrated in mitochondria. Double-labeling experiments indicated a positive correlation between the mitochondrial contents of PAG protein and glutamate, as well as between PAG enzyme activity and PAG protein as determined by Western blots. These data suggest that the antibodies recognize an enzymatically active pool of PAG. Western blots and enzyme activity assays of hippocampal homogenates revealed no change in PAG between MTLE and non-MTLE, despite a greatly (>50%) reduced number of neurons in the MTLE hippocampal formation compared to non-MTLE. Thus, the MTLE hippocampal formation contains an increased concentration and activity of PAG per neuron compared to non-MTLE. This increase suggests an enhanced capacity for glutamate synthesis-a finding that might contribute to the disrupted glutamate homeostasis in MTLE. PMID- 17115169 TI - Morphometric analysis of fibers of the human vestibular nerve: sex differences. AB - The aim of this study was to analyze myelinated axons in the human vestibular nerve (VN). We assumed that a smaller total number and average transverse area of myelinated axons in the female than male VN, would partly explain the female preponderance of vestibular disorders. The materials were obtained from 24 cadavers (12 females and 12 males) aged 54-90 years (average 74.8 years). We counted the myelinated axons, measured the transverse area of the myelinated axons, and analyzed morphological differences between the female and male specimens. The total number differed significantly between the female and male specimens. The older generation of both sexes tended to have lower total counts, but there was no significant difference among the generations. The average transverse area of the myelinated axons did not differ significantly between the female and male specimens. The older generation of both sexes tended to have a smaller average transverse area, and there was a significant difference among the generations. The presented results indicated that the lower total number, not the average transverse area, of myelinated axons in the female VN might be one of the reasons why vestibular disorders have a female preponderance. PMID- 17115170 TI - Carisoprodol intoxications: a retrospective study of forensic autopsy material from 1992-2003. AB - Carisoprodol is commonly prescribed as a centrally acting muscle relaxant, but it is also subject to abuse. The literature describing fatal intoxications with the drug is limited to a relatively small number of cases, and there are inconsistencies with regard to which concentration levels that are toxic. We therefore investigated all forensic autopsies at the Norwegian Institute of Public Health during the period 1992-2003 where carisoprodol was detected. The median concentrations of carisoprodol in intoxication with carisoprodol only or with only minor other analytical findings was 36 mg/l (range 8-65 mg/l; n=5). In the rest of the intoxications, the relevance of carisoprodol relative to the other drugs detected was variable (n=93). When the number of intoxications with carisoprodol each year were divided by the number of defined daily doses (DDD) sold, a fatal toxicity index between 5.6 and 6.9 deaths/1 million DDD was obtained. The total number of cases where carisoprodol was detected increased during the period studied, which correlated to sales figures for the drug. We conclude that carisoprodol can be fatal in concentrations below those indicated in some of the previously published literature. There were, however, only a small number of cases where the cause of death can be attributed to use of carisoprodol alone. PMID- 17115171 TI - Increase in clusterin-containing follicles in the adenohypophysis of drug abusers. AB - The hypothalamic-pituitary-adrenocortical (HPA) system in drug abusers may be affected due to disorders of the hypothalamic dopaminergic system. The present study investigated alterations in the adenohypophysis of middle-aged drug abusers (40-60 years of age), using clusterin-containing mixed cell-follicles as the indicator, in which clusterin (apolipoprotein J) is a multifunctional glycoprotein related to neurodegeneration. The paraffin-embedded adenohypophyses of methamphetamine and psychotropic drug abusers (n = 76) were compared with those of non-abusers (n = 82). The number of follicles was larger in drug abusers independent of the immediate cause of death, although the size was not significantly different. When cell types forming the follicles were immunohistochemically examined, drug abusers showed an increase of prolactin (PRL) cells and gonadotroph cells and a reciprocal decrease of growth hormone cells, suggesting hypofunction of dopaminergic neurons in the hypothalamus, while there was no change in the adrenocorticotropic hormone and thyroid-stimulating hormone cells. These increases of the clusterin-containing follicles and PRL cells in the follicles may be related to the dysfunction of dopaminergic neurons in the hypothalamus of chronic drug abusers and may be useful for investigating drug abuse in forensic casework. PMID- 17115172 TI - Fatal outcome in a child after ingestion of a transdermal fentanyl patch. AB - The case history and toxicological findings of a fatal fentanyl intoxication due to ingestion of a transdermal patch are presented. A 1-year-old otherwise healthy girl was put to bed and 2 h later she was found dead. The autopsy revealed a 25 microg/h (4.2 mg) transdermal fentanyl patch in the stomach. Toxicological analysis by liquid chromatography-tandem mass spectrometry with positive electrospray ionization yielded fentanyl and norfentanyl concentrations in the peripheral blood of 5.6 and 5.9 ng/ml, heart blood 19.0 and 8.9 ng/ml, and liver 235 and 26 ng/g, respectively. The cause of death was determined to be a fentanyl overdose. The investigation established that the child has unintentionally swallowed the patch, which had been lying on the floor. PMID- 17115173 TI - Changes in colour of different human tissues as a marker of age. AB - This study deals with age estimation based on colour changes of human tissue from the intervertebral discs, Achilles tendon and rib cartilage. The investigated colour changes are the result of the accumulation of non-enzymatic browning products in the tissue. Samples of excised tissues were photographed with a digital camera and the pictures were evaluated using the image analysis processor Lucia G 4.11 processor. The values of the intensities of the RGB channels (MeanRed, Mean Green, MeanBlue) and parameters from the IHS system (MeanSaturation, HueTypical, HueVariation, BrightVariation and MeanBrightness) were evaluated. The results confirm that colour changes of some tissues depend on ageing and are a good tool for age estimation. PMID- 17115174 TI - Successful RNA extraction from various human postmortem tissues. AB - Recently, several authors described the observation that RNA degradation does not correlate with the postmortem interval (PMI), but rather with other parameters like environmental impact and the circumstances of death. Therefore, the question arose if the analysis of gene expression could be a valuable tool in forensic genetics to contribute to the determination of the cause of death. In our study, six human tissues obtained from six individuals with PMI varying between 15 and 118 h were used for total RNA extraction. Quantification was performed using a GAPDH real-time assay, and the quality of mRNA was checked by amplification of different fragment lengths of the GAPDH transcript. In our set of samples, nearly all tissues in all PMI revealed satisfactory results, while skeletal muscle, followed by brain and heart, gave the best results. No correlation between PMI and RNA degradation could be detected, as very good results were observed for all tissues from the individual with the longest PMI. The highly promising results obtained in this study raise hopes that in the near future several fields of forensic investigation may profit from additional information about gene expression patterns and their correlation with pathological findings. PMID- 17115175 TI - Pegaptanib for myopic choroidal neovascularization in a young patient. AB - BACKGROUND: The treatment of choroidal neovascularization (CNV) due to myopic degeneration can include laser photocoagulation, photodynamic therapy, corticosteroids, and subretinal surgery. We report a case of a young patient with myopic CNV refractive to laser photocoagulation, photodynamic therapy, and intravitreal triamcinolone acetonide treated with intravitreal pegaptanib injections. METHODS: Interventional case report. The medical chart of a 36-year old female treated with intravitreal pegaptanib injections was reviewed for changes in visual acuity on the Early Treatment of Diabetic Retinopathy Study (ETDRS) chart, CNV leakage on fluorescein angiography, and adverse events reported. RESULTS: ETDRS visual acuity improved from counting fingers (CF) to 20/40 in the right eye after five, 6-weekly pegaptanib injections. CONCLUSIONS: This is the sentinel case of US Food and Drug Administration (FDA) approved pegaptanib usage for non-wet macular degeneration via an Investigational New Drug application (Title 21, Code of Federal Regulations part 132). Pegaptanib appears to be effective in treating myopic CNV refractive to laser photocoagulation, photodynamic therapy, and intravitreal triamcinolone acetonide. PMID- 17115176 TI - Effect of a 24-h continuous walking race on cardiac autonomic control. AB - This study investigated the relationships between walking speed and heart rate (HR) variability (HRV) in eleven subjects during a 24-h race. It was hypothesized that the nycthemeral rhythm on HR is preserved during the race. RR intervals and walking speed were measured. Fast Fourier transform was applied to samples of 1,024 successive RR intervals collected every hour from a HR monitor. Walking speed was averaged every hour and decreased (first lap: 8.8 +/- 0.3 vs. last lap: 7.3 +/- 0.8 km h(-1), P < 0.001) with HR also decreasing (max at 19:00 h: 143 +/- 9 vs. min at 7:00 h: 117 +/- 14 beats min(-1), P < 0.001) following a third order polynomial shape. HRV power spectral components followed distribution patterns similar to the mean RR during the race with a minimum in the early evening (19:00 20:00 h) and a maximum in the morning (5:00-8:00 h). Thus, as for mean RR, spectral components over time are also fitted to a third order polynomial regression. LF/HF ratio increased linearly (min = 0.5 +/- 0.3, max = 2.8 +/- 5.3, P = 0.02). Although mean HF peak did not decrease significantly over time, it was positively correlated with walking speed. In conclusion, this study showed that despite a constant decrease in walking speed, HR circadian rhythm is preserved during a continuous 24-h walking race. The short-term HRV components remain linked to HR whereas the LF/HF ratio increases linearly until the end of the race whatever HR is. PMID- 17115177 TI - Causes of differences in exercise-induced changes of base excess and blood lactate. AB - It has been concluded from comparisons of base excess (BE) and lactic acid (La) concentration changes in blood during exercise-induced acidosis that more H+ than La- leave the muscle and enter interstitial fluid and blood. To examine this, we performed incremental cycle tests in 13 untrained males and measured acid-base status and [La] in arterialized blood, plasma, and red cells until 21 min after exhaustion. The decrease of actual BE (-deltaABE) was 2.2 +/- 0.5 (SEM) mmol l( 1) larger than the increase of [La]blood at exhaustion, and the difference rose to 4.8 +/- 0.5 mmol l(-1) during the first minutes of recovery. The decrease of standard BE (SBE), a measure of mean BE of interstitial fluid (if) and blood, however, was smaller than the increase of [La] in the corresponding volume (delta[La](if+blood)) during exercise and only slightly larger during recovery. The discrepancy between -deltaABE and delta[La]blood mainly results from the Donnan effect hindering the rise of [La]erythrocyte to equal values like [La]plasma. The changing Donnan effect during acidosis causes that Cl- from the interstitial fluid enter plasma and erythrocytes in exchange for HCO3(-). A corresponding amount of La- remains outside the blood. SBE is not influenced by ion shifts among these compartments and therefore is a rather exact measure of acid movements across tissue cell membranes, but changes have been compared previously to delta[La]blood instead to delta[La](if+blood). When performing correct comparisons and considering Cl-/HCO3(-) exchange between erythrocytes and extracellular fluid, neither the use of deltaABE nor of deltaSBE provides evidence for differences in H+ and La- transport across the tissue cell membranes. PMID- 17115178 TI - Influence of recovery mode (passive vs. active) on time spent at maximal oxygen uptake during an intermittent session in young and endurance-trained athletes. AB - The aim of this study was to analyze the effects of recovery mode (active/passive) on time spent at high percentage of maximal oxygen uptake (VO2max) i.e. above 90% of VO2max (t90VO2max) and above 95% of VO2max (t95VO2max) during a single short intermittent session. Eight endurance-trained male adolescents (15.9 +/- 1.4 years) performed three field tests until exhaustion: a graded test to determine their VO2max (57.4 +/- 6.1 ml min(-1) kg(-1)), and maximal aerobic velocity (MAV; 17.9 +/- 0.4 km h(-1)), and in a random order, two intermittent exercises consisting of repeated 30 s runs at 105% of MAV alternated with 30 s passive (IE(P)) or active recovery (IE(A), 50% of MAV). Time to exhaustion (t(lim)) was significantly longer for IE(P) than for IE(A) (2145 +/- 829 vs. 1072 +/- 388 s, P < 0.01). No difference was found in t90VO2max and t95VO2max between IE(P) (548 +/- 499-316 +/- 360 s) and IE(A) (746 +/- 417-459 +/ 332 s). However, when expressed as a percentage of t(lim), t90VO2max and t95VO2max were significantly longer (P < 0.001 and P < 0.05, respectively) during IE(A) (67.7 +/- 19%-42.1 +/- 27%) than during IE(P) (24.2 +/- 19%-13.8 +/- 15%). Our results demonstrated no influence of recovery mode on absolute t90VO2max or t95VO2max mean values despite significantly longer t(lim) values for IE(P) than for IE(A). In conclusion, passive recovery allows a longer running time (t(lim)) for a similar time spent at a high percentage of VO2max. PMID- 17115179 TI - Development of nonexercise prediction models of maximal oxygen uptake in healthy Japanese young men. AB - The present study developed nonexercise models for predicting maximal oxygen uptake (VO2max) using skeletal muscle (SM) mass and cardiac dimensions and to investigate the validity of these equations in healthy Japanese young men. Sixty healthy Japanese men were randomly separated into two groups: 40 in the development group and 20 in the validation group. VO2max during treadmill running was measured using an automated breath-by-breath mass spectrometry system. Left ventricular internal dimensions at end-diastole (LVIDD) and at end-systole (LVIDS) were measured using M-mode ultrasound with a 2.5 MHz transducer. Stroke volume (SV) was calculated based on the Pombo rule. SM mass was predicted by B mode ultrasound muscle thickness. Correlations were observed between VO2max and predicted thigh (r = 0.74, P < 0.001) and lower leg SM mass (r = 0.55, P < 0.001). Furthermore, there were correlations between VO2max and LVIDD (r = 0.74, P < 0.001) and SV (r = 0.72, P < 0.001). Stepwise regression analysis was applied to thigh SM mass and SV for prediction of VO2max in the development group, and these parameters were closely correlated with absolute measured VO2max (R2 = 0.72, P < 0.001) by multiple regression analysis. When the VO2max prediction equations were applied to the validation group, significant correlations were also observed between the measured and predicted VO2max (R2 = 0.83, P < 0.001). These results suggested that nonexercise prediction of VO2max using thigh SM mass and cardiac dimension is a valid method to predict VO2max in young Japanese adults. PMID- 17115180 TI - Effects of glutamine and hyperoxia on pulmonary oxygen uptake and muscle deoxygenation kinetics. AB - The aim of the present study was to determine whether glutamine ingestion, which has been shown to enhance the exercise-induced increase in the tricarboxylic acid intermediate (TCAi) pool size, resulted in augmentation of the rate of increase in oxidative metabolism at the onset of exercise. In addition, the potential interaction with oxygen availability was investigated by completing exercise in both normoxic and hyperoxic conditions. Eight male cyclists cycled for 6 min at 70% VO2max following consumption of a drink (5 ml kg body mass(-1)) containing a placebo or 0.125 g kg body mass(-1) of glutamine in normoxic (CON and GLN respectively) and hyperoxic (HYP and HPG respectively) conditions. Breath-by breath pulmonary oxygen uptake and continuous, non-invasive muscle deoxygenation (via near infrared spectroscopy: NIRS) data were collected throughout exercise. The time constant of the phase II component of pulmonary oxygen uptake kinetics was unchanged between trials (CON: 21.5 +/- 3.0 vs. GLN: 18.2 +/- 1.3 vs. HYP: 18.9 +/- 2.0 vs. HPG: 18.6 +/- 1.2 s). There was also no alteration of the kinetics of relative muscle deoxygenation as measured via NIRS (CON: 5.9 +/- 0.7 vs. GLN: 7.3 +/- 0.8 vs. HYP: 6.5 +/- 0.9 vs. HPG: 5.2 +/- 0.4 s). Conversely, the mean response time of pulmonary oxygen uptake kinetics was faster (CON: 33.4 +/- 1.2 vs. GLN: 29.8 +/- 2.3 vs. HYP: 33.2 +/- 2.6 vs. HPG: 31.6 +/- 2.6 s) and the time at which muscle deoxygenation increased above pre-exercise values was earlier (CON: 9.6 +/- 0.9 vs. GLN: 8.7 +/- 1.1 vs. HYP: 8.5 +/- 0.8 vs. HPG: 8.4 +/- 0.7 s) following glutamine ingestion. In normoxic conditions, plasma lactate concentration was lower following glutamine ingestion compared to placebo. Whilst the results of the present study provide some support for the present hypothesis, the lack of any alteration in the time constant of pulmonary oxygen uptake and muscle deoxygenation kinetics suggest that the normal exercise induced expansion of the TCAi pool size is not limiting to oxidative metabolism at the onset of cycle exercise at 70% VO2max. PMID- 17115181 TI - Eye blink frequency during different computer tasks quantified by electrooculography. AB - The purpose of the study was to evaluate electrooculography (EOG) as an automatic method to measure the human eye blink frequency (BF) during passive and interactive computer tasks performed at two screen heights. Ten healthy subjects (5 males and 5 females) participated in the study in a 23 degrees C temperature and 30-35% relative humidity controlled simulated office environment. Each test subject completed a 2 x 10 min active task of computer work and a 3 x 10 min passive task of watching a film on a video display unit (VDU). Both tasks included two viewing angles: standard (the monitors' upper edge was in the same height as the subjects' eyes) and low (lowered by 25 degrees). EOG signals were recorded with two Ag/AgCl surface electrodes positioned above and below the right eye, and a reference electrode was placed behind the ear. The experiments were video filmed, and eye blinks were counted manually from the video recordings and compared to the EOG measurements. The method showed a high validity to detect blinks during computer work: 95.4% of the blinks were retrieved by the EOG method and very few artefacts from eye movements were erroneously classified as eye blinks (2.4%). By use of the EOG method, the computer task was found to significantly decrease the BF by 69% compared to the passive task (P < 0.001), and a small decrease (12-14%) was found by lowering the viewing angle by 25 degrees. PMID- 17115182 TI - Functional diversity of tocochromanols in plants. AB - Tocochromanols encompass a group of compounds with vitamin E activity essential for human nutrition. They accumulate in photooxidative organisms, e.g. in some algae and in plants, where they localize to thylakoid membranes and plastoglobules of chloroplasts. Tocochromanols contain a polar chromanol head group with a long isoprenoid side chain. Depending on the nature of the isoprenoid chain, tocopherols (containing a phytyl chain) or tocotrienols (geranylgeranyl chain) can be distinguished in plants. The tocochromanol biosynthetic pathway has been studied in Arabidopsis and Synechocystis in recent years, and the respective mutants and genes were isolated. Mutant characterization revealed that tocopherol protects lipids in photosynthetic membranes and in seeds against oxidative stress. In addition to its antioxidant characteristics, tocopherol was shown be involved in non-antioxidant functions such as primary carbohydrate metabolism. A considerable proportion of tocopherol is synthesized from free phytol suggesting that excess amounts of phytol released from chlorophyll breakdown during stress or senescence might be deposited in the form of tocopherol in chloroplasts. PMID- 17115183 TI - Acquired torticollis as the only initially presenting symptom in a child with a brainstem glioma. PMID- 17115184 TI - Oral purified bacterial extracts in acute respiratory tract infections in childhood: a systematic quantitative review. AB - BACKGROUND: Recurrent acute respiratory tract infections (ARTI) are a common problem in childhood. Some evidence suggests a benefit regarding the prevention of ARTI in children treated with the immunomodulator OM-85 BV (Bronchovaxom). METHODS: We summarised the evidence on the effectiveness of the immunomodulator OM-85 BV in the prevention of ARTI in children. We searched randomised comparisons of oral purified bacterial extracts against inactive controls in children with respiratory tract diseases in nine electronic databases and reference lists of included studies. We extracted salient features of each study, calculated relative risks (RR) or weighted mean differences (WMD) and performed meta-analyses using random-effects models. RESULTS: Thirteen studies (2,721 patients) of low to moderate quality tested OM-85 BV. Patients and outcomes differed substantially, which impeded pooling results of more than two trials. Two studies (240 patients) reporting on the number of patients with less than three infections over 6 month of follow-up in children not in day care showed a trend for benefit RR 0.82 (95% CI, 0.65-1.02). One out of two studies examining the number of children not in day care without infections over 4-6 month reported a significant RR of 0.42 (95% CI, 0.21-0.82) whereas the smaller, second study did not [RR 0.92 (95% CI, 0.58-1.46)]. Two studies reporting the number of antibiotic courses indicated a benefit for the intervention arm [WMD 2.0 (95% CI, 1.7-2.3)]. Two out of the three studies showed a reduction of length of episodes of 4-6 days whereas a third study showed no difference between the two groups. CONCLUSION: Evidence in favour of OM-85 BV in the prevention of ARTI in children is weak. There is a trend for fewer and shorter infections and a reduction of antibiotic use. PMID- 17115185 TI - Functional inference of the methylenetetrahydrofolate reductase 677C > T and 1298A > C polymorphisms from a large-scale epidemiological study. AB - Two functional single nucleotide polymorphisms, 677C > T and 1298A > C have been described for the methylenetetrahydrofolate (MTHFR) gene. Both are associated with reduced enzyme activity in vitro. For the 677T, but not the 1298C allele, significantly lower serum folate and higher plasma total homocysteine (tHcy) have been reported. We genotyped 10,034 middle-aged (50-64 years old) subjects and measured serum folate and tHcy. Within strata of 677 genotypes, 1,298 genotypes had significantly different serum folate and tHcy (P < or = 0.03 for all comparisons). Each additional 1298C allele reduced mean serum folate and increased mean tHcy, by (on average) 4.5 and 3.0%, respectively. In comparison, within strata of 1,298 genotypes, the increase from no, to one 677T-allele reduced serum folate and increased tHcy by, 7.1 and 6.3%, respectively. Lowest serum folate and highest tHcy level was found for the 677TT/1298AA genotype. The difference in tHcy was significantly larger at low folate than at high folate when genotypes 677TT/1298AA and 677CT/1298AA, 677CT/1298AC and 677CC/1298AC, and genotypes 677CT/1298AC and 677CT/1298AA were compared. We interpreted these data in the context of a model of the MTHFR enzyme that describes the enzyme as a dimer that mainly exist in six different configurations. The model reconciled the observed phenotypic effects of the 677/1,298 combination genotypes with previous in vitro measurements, and identified enzyme configurations that are sensitive to low folate levels. In conclusion, this report demonstrates functional inference of the MTHFR 677 C > T and 1,298 A > C polymorphisms from a large-scale epidemiological study. PMID- 17115186 TI - TagSNP evaluation for the association of 42 inflammation loci and vascular disease: evidence of IL6, FGB, ALOX5, NFKBIA, and IL4R loci effects. AB - Inflammatory markers have consistently been associated with vascular disease. Evidence of genetic polymorphisms in inflammatory loci that predict severe carotid artery disease (CAAD) would suggest that this relationship is not secondary to other correlated factors, but related to inflammation itself. We examined the full common genetic variation in 42 inflammatory loci for prediction of severe CAAD versus ultrasound proven controls using a tagSNP approach. For selected loci, monocyte RNA levels were contrasted in subjects with and without CAAD. We confirm the association of IL6(-174), FGB (-455), and ALOX5 with CAAD and show that multiple ALOX5 SNPs independently predict CAAD. We provide evidence for previously unreported associations of SNPs in IL4R, NFKBIA, and PLG with CAAD, and weaker evidence for associations with CSF3, IL10RA, and VCAM1. The NFKBIA and IL10RA expression levels significantly differed between subjects with CAAD and controls. These results support a role for genetic variation related to inflammation in CAAD and a causal role for specific gene products. PMID- 17115187 TI - A genome-wide linkage scan for quantitative trait loci underlying obesity related phenotypes in 434 Caucasian families. AB - To identify quantitative trait loci (QTLs) that contribute to obesity, we performed a large-scale whole genome linkage scan (WGS) involving 4,102 individuals from 434 Caucasian families. The most pronounced linkage evidence was found at the genomic region 20p11-12 for fat mass (LOD = 3.31) and percentage fat mass (PFM) (LOD = 2.92). We also identified several regions showing suggestive linkage signals (threshold LOD = 1.9) for obesity phenotypes, including 5q35, 8q13, 10p12, and 17q11. PMID- 17115188 TI - Extremely skewed X-chromosome inactivation is increased in pre-eclampsia. AB - Pre-eclampsia is a disorder that affects approximately 5% of pregnancies. We tested the hypothesis that skewed X-chromosome inactivation (XCI) could be involved in the pathogenesis of pre-eclampsia. Peripheral blood DNA was obtained from 67 pre-eclampsia patients and 130 control women. Androgen receptor (AR) was analyzed by the HpaII/polymerase chain reaction assay to assess XCI patterns in DNA extracted from peripheral-blood cells. In addition, buccal cells were obtained from seven patients, and the analysis repeated. Extremely skewed XCI was observed in 10 of 46 informative patients (21.74%), and in 2 of 86 informative controls (2.33%, P = 0.0005; chi(2) test). Our findings support a role for the X chromosome in the pathogenesis of pre-eclampsia in a subgroup of patients. PMID- 17115189 TI - Seasonal, daily and diurnal variations in the stable carbon isotope composition of carbon dioxide respired by tree trunks in a deciduous oak forest. AB - The stable C isotope composition (delta13C) of CO2 respired by trunks was examined in a mature temperate deciduous oak forest (Quercus petraea). Month-to month, day-to-day and diurnal, measurements were made to determine the range of variations at different temporal scales. Trunk growth and respiration rates were assessed. Phloem tissue was sampled and was analysed for total organic matter and soluble sugar 13C composition. The CO2 respired by trunk was always enriched in 13C relative to the total organic matter, sometimes by as much as 5 per thousand. The delta13C of respired CO2 exhibited a large seasonal variation (3.3 per thousand), with a relative maximum at the beginning of the growth period. The lowest values occurred in summer when the respiration rates were maximal. After the cessation of radial trunk growth, the respired CO2 delta13C values showed a progressive increase, which was linked to a parallel increase in soluble sugar content in the phloem tissue (R=0.95; P<0.01). At the same time, the respiration rates declined. This limited use of the substrate pool might allow the discrimination during respiration to be more strongly expressed. The late-season increase in CO2 delta13C might also be linked to a shift from recently assimilated C to reserves. At the seasonal scale, CO2 delta13C was negatively correlated with air temperature (R=-0.80; P<0.01). The diurnal variation sometimes reached 3 per thousand, but the range and the pattern depended on the period within the growing season. Contrary to expectations, diurnal variations were maximal in winter and spring when the leaves were missing or not totally functional. By contrast to the seasonal scale, these diurnal variations were not related to air temperature or sugar content. Our study shows that seasonal and diurnal variations of respired 13C exhibited a similar large range but were probably explained by different mechanisms. PMID- 17115190 TI - Physical background of the development of oxygen depletion in ice-covered lakes. AB - The effect of the heat interaction between a water column and sediments on the formation, development, and duration of existence of anaerobic zones in ice covered lakes is estimated based on observational data from five frozen lakes located in northwestern Russia and North America. A simple one-dimensional model that describes the formation and development of the dissolved oxygen deficit in shallow ice-covered lakes is suggested. The model reproduces the main features of dissolved oxygen dynamics during the ice-covered period; that is, the vertical structure, the thickness, and the rate of increase of the anaerobic zone in bottom layers. The model was verified against observational data. The results from the verification show that the model adequately describes the dissolved oxygen dynamics in winter. The consumption rates of DO by bacterial plankton and by bottom sediments, which depend on the heat transfer through the water-sediment interface, are calculated. The results obtained allow the appearance of potentially dangerous anaerobic zones in shallow lakes and in separate lake areas, which result from thermal regime changes, to be predicted. PMID- 17115192 TI - Treatment of steroid-resistant membranous lupus nephritis with plasmapheresis and low-dose cyclosporine. PMID- 17115193 TI - Developmental origins of adult hypertension: new insights into the role of the kidney. AB - It is now accepted that early life environment can modulate adult phenotype. One of the best documented examples is the effect of prenatal environment on adult hypertension and cardiovascular morbidity. Human epidemiologic studies have been complemented with experimental models showing, for example, that maternal dietary manipulations during pregnancy in the rat can be used to induce adult hypertension in the offspring. The weight of the emerging evidence suggests that abnormal Na handling by the kidney plays an important role in the pathogenesis of the hypertension. Although the number of nephrons is modestly reduced in most experimental models, there is very little change in total glomerular filtration rate, casting doubt on the hypothesis that restricted Na filtration is the major mechanism. Recent studies have instead strongly suggested that renal tubular handling of Na is altered, resulting in an altered set-point for Na balance. The mechanism may involve intrarenal inflammation and increased oxidative stress which disrupt the tubulointerstitial microenvironment, leading to constitutively upregulated Na reabsorption in the distal tubule. The upregulation may be mediated by autocrine and paracrine factors promoting distal tubule Na reabsorption. A similar mechanism has been hypothesized to be important in other types of hypertension and may hence be a common pathway in the genesis of volume dependent hypertension. PMID- 17115194 TI - Hyponatremia resulting from arginine vasopressin receptor 2 gene mutation. AB - Chronic hyponatremia, unless associated with extracellular fluid volume expansion, is an infrequent electrolyte imbalance in pediatrics. We report an infant with chronic hyponatremia suggestive of a syndrome of inappropriate secretion of antidiuretic hormone (SIADH), in the absence of ADH secretion. A mutation was found in the same codon of the gene that results in a loss-of- function of arginine vasopressin receptor 2 (AVPR2) observed in congenital nephrogenic diabetes insipidus. In this case, a gain-of- function of AVPR 2 was found to be responsible for a SIADH-like state. PMID- 17115195 TI - Blood pressure control in pediatric hemodialysis: the Midwest Pediatric Nephrology Consortium Study. AB - Hypertension is frequent in pediatric patients receiving dialysis, with an especially high rate reported in children on hemodialysis (HD). We performed the present study to assess blood pressure (BP) status and identify risk factors for poor BP control in children on maintenance HD. One month's dialysis records were collected from 71 subjects receiving HD in ten dialysis units participating in the Midwest Pediatric Nephrology Consortium (MWPNC). For each HD session, data on pre- and posttreatment weights and BPs were recorded. Hypertension, defined as mean BP >or= 95th percentile, was found in 42 (59%) subjects. Eleven subjects (15.5%) had prehypertension, defined as mean BP between the 90th and 95th percentiles, while 18 subjects (25.3%) had normal BP (<90th percentile). BP significantly decreased at the end of a dialysis session; however, only 15 of 42 hypertensive subjects (35%) normalized their BP. Hypertensive subjects were younger (p = 0.03), had higher serum phosphorus (p = 0.01), and had more elevated posttreatment weight above estimated dry weight (p = 0.02). Logistic regression showed that younger age (p = 0.02) and higher serum phosphorus (p = 0.02) independently predicted hypertensive status. In conclusion, this study emphasizes the difficulty of BP control in pediatric HD patients. Especially poor BP control was found in younger children; those patients who do not reach their posttreatment weight goals, perhaps reflecting their hypervolemic state; and those who have higher serum phosphorus levels. PMID- 17115196 TI - High-sensitivity C-reactive protein (hs-CRP) level in children with nephrotic syndrome. AB - The aim of this study was to assess the serum concentration of high-sensitivity C reactive protein (hs-CRP) in children with nephrotic syndrome (NS) treated with prednisone and cyclosporine A (CyA). Patients were divided into three groups: (I) 20 NS children (aged 4-14 years) in relapse and examined twice, (A) before treatment and (B) after proteinuria regression (a 3-4 week course of prednisone therapy); (II) 20 children with steroid-dependent or steroid-resistant NS, treated with CyA, also examined twice, (D) before treatment with CyA, (E) 6 months after therapy. A control group (C) consisted of 20 healthy children. Serum hs-CRP level was determined by a nephelometric method with a Behring Nephelometer 100 Analyzer, Dade Behring. The results showed that median hs-CRP concentration was the highest in children with relapsing steroid-sensitive NS before treatment (IA). After proteinuria regression (IB), the hs-CRP level had decreased and did not differ from that of healthy controls (C) (P>0.05). In group II, before CyA administration (IID), the level of hs-CRP was normal, but it had increased after 6 months of treatment (IIE) up to a level six-times higher than that of the control group (P<0.01). We concluded that, in children with steroid-sensitive nephrotic syndrome in relapse, the serum hs-CRP level is increased but returns to normal after 3-4 weeks of glucocorticoid treatment. In children chronically treated with CyA due to NS, serum hs-CRP level increases significantly during the therapy. PMID- 17115198 TI - Xanthinuria type I: a rare cause of urolithiasis. AB - Xanthinuria type I is a rare disorder of purine metabolism caused by xanthine oxidoreductase or dehydrogenase (XDH) deficiency. We report a family with two affected children out of 335 pediatric stone patients studied since 1991 in Armenia. The propositus, a 13-month-old boy, presented with abdominal pain and urinary retention followed by stone passage (0.9x0.6 cm). Infrared spectroscopy in Yerevan revealed a pure xanthine stone. Family examination in the parents and brother was normal, but the propositus and his 8-year-old asymptomatic sister had hypouricemia, hypouricosuria, and high urinary excretion of hypoxanthine and xanthine. Ultrasonography in the index patient showed bilateral stones requiring pyelolithotomy. High fluid intake and purine restriction did not prevent further stone passages. The affected asymptomatic sister had a small pelvic stone (4 mm). Mutation analysis revealed a heterozygous novel base pair substitution in exon 25 of the XDH gene (c.2810C>T), resulting in an amino acid substitution (p.Thr910Met). The second mutation could not be detected. Despite this, the heterozygous mutation, the chemical findings, and the positive allopurinol test altogether prove xanthinuria type I, which may present wide clinical intrafamilial variation. Diagnosis is suspected usually from low serum uric acid. No specific therapy is available. PMID- 17115199 TI - Lack of evidence of hypervolemia in children with insulin-dependent diabetes mellitus. AB - Hypervolemia is considered to play a major role in the pathogenesis of diabetic vasculo- and nephropathy. The aim of our study is to determine whether children and adolescents with insulin-dependent diabetes mellitus (IDDM) experience alterations in blood volume (BV) before onset of apparent nephropathy. BV (calculated as the sum of measured plasma volume (PV) and red cell volume (RCV)) was determined in 31 children (9-16 yr) with a mean duration of IDDM of 6.6 yr and without microalbuminuria. Due to dependence of these values on age, size and sex, all data were normalised for body size parameters. While no statistical difference for BV normalised for lean body mass (LBM) (86.98+/-9.5 ml/kg) was found in diabetic children compared with our control population (84.91+/-12.08 ml/kg), a difference could be shown when normalised for body surface area (BSA) (diabetic children 2.37+/-0.3 L/m(2); control population 2.15+/-0.38 L/m(2), p=0.002). Increased BV is only present when normalising for BSA and not for the theoretical superior LBM-index. Because the study population exhibited a poor glycemic control (HbA1c 10.2+/-2.4 %), an influence of glucosuria-induced polyuria on BV cannot be excluded. Taking into account these limitations our data do not confirm the presence of hypervolemia before onset of diabetic nephropathy. PMID- 17115200 TI - Clinicians' practice and attitudes toward cancer pain management in Korea. AB - OBJECTIVES: The purposes of this study were to assess clinicians' (n = 250) current practices and attitudes about cancer pain management and to identify perceived concerns about and barriers to pain control in urban cancer-treatment settings in Korea. MATERIALS AND METHODS: Survey data (20 items) were collected either by mail or interview. Approximately 40% of the sample were nurses. More medical clinicians than surgical clinicians had more than 5 years of experience with cancer care (43 vs 31%) and committed more than 30% of their working hours to palliation (61 vs 19%). Significantly more medical clinicians claimed to be somewhat more or much more liberal than their professional peers in using analgesics compared with surgical clinicians (54 vs 35%). The liberal sample was more likely to be male (OR = 3.3, p < 0.001) and to be more experienced with cancer care (OR = 1.2, p < 0.001). Medical clinicians also reported more adequate pain-management training and a more proactive approach to assessing and treating pain. RESULTS: Overall, the greatest concerns regarding opioid use were safety, side effects, and fear of addiction. Inadequate pain assessment and lack of staff knowledge and time were identified as barriers to pain management. Unrealistic expectations and denial from both patient and family were the most troublesome issues for delivery of care to dying patients. This study suggests a more conservative attitude toward cancer pain management in Korea than in other countries surveyed in a similar manner. CONCLUSION: A combination of routine professional education and dissemination of guidelines is needed to bring about significant improvement in cancer pain control in Korea. PMID- 17115201 TI - Paxillus involutus mycorrhiza attenuate NaCl-stress responses in the salt sensitive hybrid poplar Populusxcanescens. AB - In order to characterise the effect of ectomycorrhiza on Na+-responses of the salt-sensitive poplar hybrid Populus x canescens, growth and stress responses of Paxillus involutus (strain MAJ) were tested in liquid cultures in the presence of 20 to 500 mM NaCl, and the effects of mycorrhization on mineral nutrient accumulation and oxidative stress were characterised in mycorrhizal and non mycorrhizal poplar seedlings exposed to 150 mM NaCl. Paxillus involutus was salt tolerant, showing biomass increases in media containing up to 500 mM NaCl after 4 weeks growth. Mycorrhizal mantle formation on poplar roots was not affected by 150 mM NaCl. Whole plant performance was positively affected by the fungus because total biomass was greater and leaves accumulated less Na+ than non mycorrhizal plants. Energy dispersive X-ray microanalysis using transmission electron microscopy analysis of the influence of mycorrhization on the subcellular localisation of Na+ and Cl- in roots showed that the hyphal mantle did not diminish salt accumulation in root cell walls, indicating that mycorrhization did not provide a physical barrier against excess salinity. In the absence of salt stress, mycorrhizal poplar roots contained higher Na+ and Cl- concentrations than non-mycorrhizal poplar roots. Paxillus involutus hyphae produced H2O2 in the mantle but not in the Hartig net or in pure culture. Salt exposure resulted in H2O2 formation in cortical cells of both non-mycorrhizal and mycorrhizal poplar and stimulated peroxidase but not superoxide dismutase activities. This shows that mature ectomycorrhiza was unable to suppress salt induced oxidative stress. Element analyses suggest that improved performance of mycorrhizal poplar under salt stress may result from diminished xylem loading of Na+ and increased supply with K+. PMID- 17115202 TI - The association between cervical spine curvature and neck pain. AB - Degenerative changes of the cervical spine are commonly accompanied by a reduction or loss of the segmental or global lordosis, and are often considered to be a cause of neck pain. Nonetheless, such changes may also remain clinically silent. The aim of this study was to examine the correlation between the presence of neck pain and alterations of the normal cervical lordosis in people aged over 45 years. One hundred and seven volunteers, who were otherwise undergoing treatment for lower extremity problems in our hospital, took part. Sagittal radiographs of the cervical spine were taken and a questionnaire was completed, enquiring about neck pain and disability in the last 12 months. Based on the latter, subjects were divided into a group with neck pain (N = 54) and a group without neck pain (N = 53). The global curvature of the cervical spine (C2-C7) and each segmental angle were measured from the radiographs, using the posterior tangent method, and examined in relation to neck complaints. No significant difference between the two groups could be found in relation to the global curvature, the segmental angles, or the incidence of straight-spine or kyphotic deformity (P > 0.05). Twenty-three per cent of the people with neck pain and 17% of those without neck pain showed a segmental kyphosis deformity of more than 4 degrees in at least one segment--most frequently at C4/5, closely followed by C5/6 and C3/4. The average segmental angle at the kyphotic level was 6.5 degrees in the pain group and 6.3 degrees in the group without pain, with a range of 5-10 degrees in each group. In the group with neck pain, there was no association between any of the clinical characteristics (duration, frequency, intensity of pain; radiating pain; sensory/motor disturbances; disability; healthcare utilisation) and either global cervical curvature or segmental angles. The presence of such structural abnormalities in the patient with neck pain must be considered coincidental, i.e. not necessarily indicative of the cause of pain. This should be given due consideration in the differential diagnosis of patients with neck pain. PMID- 17115203 TI - On the use of denaturing gradient gel electrophoresis approach for bacterial identification in endodontic infections. AB - Bacteria in infected root canals of teeth evincing chronic apical periodontitis lesions were identified by a polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) approach. DNA was extracted from root canal samples, and part of the 16S rRNA gene of all bacteria was amplified by PCR and separated by DGGE, generating banding patterns representative of the community structure. Twenty visible bands were cut out of the gel, re-amplified, and sequenced to provide identification. Sequencing analysis revealed the presence of both cultivable and as-yet-uncultivated species in the samples analyzed, including representatives of the genera Fusobacterium, Bacteroides, Dialister, Synergistes, Prevotella, Eubacterium and Peptostreptococcus. Unambiguous identification was not always possible and the method's limitations are discussed. In general, the findings showed that PCR-DGGE can be useful for the identification of both cultivable and as-yet-uncultivated bacteria in endodontic infections. PMID- 17115204 TI - Neuroradiological findings in vascular dementia. AB - INTRODUCTION: There are multiple diagnostic criteria for vascular dementia (VaD) that may define different populations. Utilizing the criteria of the National Institute of Neurological Disorders and Stroke and Association Internationale pour la Recherche et l'Enseignement en Neurosciences (NINDS-AIREN) has provided improved consistency in the diagnosis of VaD. The criteria include a table listing brain imaging lesions associated with VaD. METHODS: The different neuroradiological aspects of the criteria are reviewed based on the imaging data from an ongoing large-scale clinical trial testing a new treatment for VaD. The NINDS-AIREN criteria were applied by a centralized imaging rater to determine eligibility for enrollment in 1,202 patients using brain CT or MRI. RESULTS: Based on the above data set, the neuroradiological features that are associated with VaD and that can result from cerebral small-vessel disease with extensive leukoencephalopathy or lacunae (basal ganglia or frontal white matter), or may be the consequence of single strategically located infarcts or multiple infarcts in large-vessel territories, are illustrated. These features may also be the consequence of global cerebral hypoperfusion, intracerebral hemorrhage, or other mechanisms such as genetically determined arteriopathies. CONCLUSION: Neuroimaging confirmation of cerebrovascular disease in VaD provides information about the topography and severity of vascular lesions. Neuroimaging may also assist with the differential diagnosis of dementia associated with normal pressure hydrocephalus, chronic subdural hematoma, arteriovenous malformation or tumoral diseases. PMID- 17115205 TI - High level expression of a recombinant phospholipase C from Bacillus cereus in Bacillus subtilis. AB - Twenty-two Bacillus cereus strains were screened for phospholipase C (PLC, EC 3.1.4.3) activity using p-nitrophenyl phosphorylcholine as a substrate. Two strains (B. cereus SBUG 318 and SBUG 516) showed high activity at elevated temperatures (>70 degrees C) at acidic pH (pH 3.5-6) and were selected for cloning and functional expression using Bacillus subtilis. The genes were amplified from B. cereus DNA using primers based on a known PLC sequence and cloned into the expression vector pMSE3 followed by transformation into B. subtilis WB800. On the amino acid level, one protein (PLC318) was identical to a PLC described from B. cereus, whereas PLC516 contained an amino acid substitution (E173D). PLC production using the recombinant strains was performed by an acetoin controlled expression system. For PLC516, 13.7 U g(-1) wet cell weight was determined in the culture supernatant after 30 h cultivation time. Three purification steps resulted in pure PLC516 with a specific activity of 13,190 U mg(-1) protein. PMID- 17115206 TI - Membrane biofouling by extracellular polymeric substances or soluble microbial products from membrane bioreactor sludge. AB - This study extracted the soluble microbial products and loosely bound and tightly bound extracellular polymeric substances (EPS) from suspended sludge from a membrane bioreactor, original and aerobically/anaerobically digested, and compared their fouling potentials on a microfiltration membrane. The resistance of cake layer accounts for 95-98% of the total filtration resistances when filtering the whole sludges, with anaerobically digested sludge presenting the highest resistance among the three tested sludges. The tightly bound EPS has the highest potential to foul the membrane; however, the loosely bound EPS contribute most of the filtration resistances of the whole sludges. The foulants corresponding to the irreversible fouling have chemical fingerprints similar to those from loosely bound EPS, which have a greater predilection to proteins and humic substances than to polysaccharides. PMID- 17115207 TI - Comparison of methods for total community DNA extraction and purification from compost. AB - The differences on DNA yield and purity of three different DNA extraction protocols were compared with regard to the use for PCR and other molecular analyses. Total DNA was extracted from compost by the three protocols, and then was purified by spin-bind cartridges after being precipitated by PEG8000. The detection performed on a nucleic acid and protein analyzer showed that all three methods produced high DNA yields. The agarose gel electrophoresis showed that the fragments of crude and purified DNA had a length of about 23 kb. A eubacterial 16S rRNA gene-targeted primer pair was used for PCR amplification, and full length 16S rDNAs were amplified from all the purified DNA samples. After being digested by restriction endonucleases, the restriction map of amplified rDNA showed identical genetic diversity. The products of PCR using primer pair GC341F and 907R were also used for denaturing gradient gel electrophoresis analysis. The results indicated that high-quality DNA was extracted from compost by the three protocols, and each of the protocols is adapted to extract microbial genome DNA from compost expediently and cheaply. PMID- 17115208 TI - Molecular cloning and heterologous expression of a new xylanase gene from Plectosphaerella cucumerina. AB - A gene encoding a new xylanase, named xynZG, was cloned by the genome-walking PCR method from the nematophagous fungus Plectosphaerella cucumerina. The genomic DNA sequence of xynZG contains a 780 bp open reading frame separated by two introns with the sizes of 50 and 46 bp. To our knowledge, this would be the first functional gene cloned from P. cucumerina. The 684 bp cDNA was cloned into vector pHBM905B and transformed into Pichia pastoris GS115 to select xylanase-secreting transformants on RBB-xylan containing plate. The optimal secreting time was 3 days at 25 degrees C and enzymatic activities in the culture supernatants reached the maximum level of 362 U ml(-1). The molecular mass of the enzyme was estimated to be 19 kDa on SDS-PAGE. The optimal pH and temperature of the purified enzyme is 6 and 40 degrees C, respectively. The purified enzyme is stable at room temperature for at least 10 h. The Km and Vmax values for birchwood xylan are 2.06 mg ml(-1) and 0.49 mmol min(-1)mg(-1), respectively. The inhibitory effects of various mental ions were investigated. It is interesting to note that Cu2+ ion, which strongly inhibits most other xylanases studied, reduces enzyme activity by only 40%. Furthermore, enzyme activity is unaffected by EDTA even at a concentration of 5 mM. PMID- 17115209 TI - Engineering the lycopene synthetic pathway in E. coli by comparison of the carotenoid genes of Pantoea agglomerans and Pantoea ananatis. AB - The lycopene synthetic pathway was engineered in Escherichia coli using the carotenoid genes (crtE, crtB, and crtI) of Pantoea agglomerans and Pantoea ananatis. E. coli harboring the P. agglomerans crt genes produced 27 mg/l of lycopene in 2YT medium without isopropyl-beta-D: -thiogalactopyranoside (IPTG) induction, which was twofold higher than that produced by E. coli harboring the P. ananatis crt genes (12 mg/l lycopene) with 0.1 mM IPTG induction. The crt genes of P. agglomerans proved better for lycopene production in E. coli than those of P. ananatis. The crt genes of the two bacteria were also compared in E. coli harboring the mevalonate bottom pathway, which was capable of providing sufficient carotenoid building blocks, isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), with exogenous mevalonate supplementation. Lycopene production significantly increased using the mevalonate bottom pathway and 60 mg/l of lycopene was obtained with the P. agglomerans crt genes, which was higher than that obtained with the P. ananatis crt genes (35 mg/l lycopene). When crtE among the P. ananatis crt genes was replaced with P. agglomerans crtE or Archaeoglobus fulgidus gps, both lycopene production and cell growth were similar to that obtained with P. agglomerans crt genes. The crtE gene was responsible for the observed difference in lycopene production and cell growth between E. coli harboring the crt genes of P. agglomerans and P. ananatis. As there was no significant difference in lycopene production between E. coli harboring P. agglomerans crtE and A. fulgidus gps, farnesyl diphosphate (FPP) synthesis was not rate-limiting in E. coli. PMID- 17115210 TI - A chemostat study of Streptomyces peucetius var. caesius N47. AB - The behavior of Streptomyces peucetius var. caesius N47 was studied in a glucose limited chemostat with a complex cultivation medium. The steady-state study yielded the characteristic constants mu (max) over 0.10 h(-1), Y (XS) 0.536 g g( 1), and m(S) 0.54 mg g(-1) h(-1). The product of secondary metabolism, epsilon rhodomycinone, was produced with characteristics Y (PX) 12.99 mg g(-1) and m (P) 1.20 mg g(-1) h(-1). Significant correlations were found for phosphate and glucose consumption with biomass and epsilon-rhodomycinone production. Metabolic flux analysis was conducted to estimate intracellular fluxes at different dilution rates. TCA, PPP, and shikimate pathway fluxes exhibited bigger values with production than with growth. Environmental perturbation experiments with temperature, airflow, and pH changes on a steady-state chemostat implied that an elevation of pH could be the most effective way to shift the cells from growing to producing, as the pH change induced the biggest transient increase to the calculated epsilon-rhodomycinone flux. PMID- 17115211 TI - High level expression of human endostatin in Pichia pastoris using a synthetic gene construct. AB - Endostatin, a 20-kDa C-terminal fragment derived from type XVIII collagen, is a potent angiogenesis inhibitor and an antitumor factor. To improve the production of recombinant human endostatin on increasing demand in clinical practice, we constructed an artificial gene encoding its mature peptide sequence in human collagen XVIII. The synthetic gene consisted of 20 codons in preference in methylotropic yeast-Pichia pastoris and was cloned into expression vector pPICZalphaA; and the recombinant protein was expressed in P. pastoris strain SMD1168 and purified to near homogeneity using heparin affinity chromatography. The amount of expressed recombinant protein in cultural media using described strategy was 80 mg/l in shake flask cultivation and 435 mg/l in high-density bioreactor fermentation. Methylthiazolium assay demonstrated that human endostatin expressed in P. pastoris using artificial synthetic gene of preference in P. pastoris was able to inhibit the acidic fibroblast growth factor-induced proliferation of endothelial cells in vitro. PMID- 17115212 TI - Biosynthesis and engineering of isoprenoid small molecules. AB - Isoprenoid secondary metabolites are a rich source of commercial products that have not been fully explored. At present, there are isoprenoid products used in cancer therapy, the treatment of infectious diseases, and crop protection. All isoprenoids share universal prenyl diphosphate precursors synthesized via two distinct pathways. From these universal precursors, the biosynthetic pathways to specific isoprenoids diverge resulting in a staggering array of products. Taking advantage of this diversity has been the focus of much effort in metabolic engineering heterologous hosts. In addition, the engineering of the mevalonate pathway has increased levels of the universal precursors available for heterologous production. Finally, we will describe the efforts to produce to commercial terpenoids, paclitaxel and artemisinin. PMID- 17115213 TI - CT manifestations of ileal dysgenesis. AB - Ileal dysgenesis is an uncommon condition of unknown etiology occurring in the distal ileum in the region of the vitelline duct. The CT appearance of this lesion, although not previously described to our knowledge, is characteristic. We report a patient with ileal dysgenesis who had an abdominal CT scan to evaluate chronic iron deficiency anemia and protein-losing enteropathy. Recognition of this lesion by pediatric radiologists is important; so that surgical treatment, which is simple and effective, can be initiated quickly. PMID- 17115214 TI - Imaging atherosclerosis in the vulnerable patient. PMID- 17115215 TI - A hybrid algorithm for PET/CT image merger in hybrid scanners. AB - PURPOSE: To improve the PET image quality of a hybrid PET/CT scanner by merging CT borders with PET texture. PET/CT scanners provide both high-resolution CT images showing anatomical details and PET images of low-resolution physiological information about radiopharmaceutical uptake. Standard smoothing of noisy PET images may further impair PET resolution, reducing small lesion detectability. METHODS: The CT edge data and the PET texture data were merged using a modified form of an algorithm called HCT (hybrid computed tomography). In merged PET/CT images, each PET pixel value was estimated by iteratively applying a corrected 2D Taylor expansion to each of its eight neighbors. The spatial derivative term was used only near anatomical edges provided by the CT. This counts-preserving algorithm was tested on a special resolution phantom and patient data sets obtained by PET/CT acquisitions. RESULTS: The HCT algorithm provided phantom PET images with sharp borders and improved resolution (< or = 3 mm as compared to > or = 4 mm). HCT increased the signal to background contrast ratios by an average of 61% (40-89%) while maintaining noise reduction similar to the Gaussian filtering standard in PET. In the clinical PET images, HCT allowed for an improved delineation of pulmonary and pelvic lesions and an improved visualization of the brain. CONCLUSION: A new reconstruction algorithm for merging CT anatomical edge data with functional PET data has been introduced. The algorithm smooths noisy PET images while retaining sharper edges at corresponding anatomical borders, resulting in an improvement in resolution and contrast ratio. PMID- 17115216 TI - Experiments and models of sensorimotor interactions during locomotion. AB - During locomotion sensory information from cutaneous and muscle receptors is continuously integrated with the locomotor central pattern generator (CPG) to generate an appropriate motor output to meet the demands of the environment. Sensory signals from peripheral receptors can strongly impact the timing and amplitude of locomotor activity. This sensory information is gated centrally depending on the state of the system (i.e., rest vs. locomotion) but is also modulated according to the phase of a given task. Consequently, if one is to devise biologically relevant walking models it is imperative that these sensorimotor interactions at the spinal level be incorporated into the control system. PMID- 17115217 TI - Modeling the pre- and post-synaptic components involved in the synaptic modification between cones and horizontal cells in carp retina. AB - In retinal synapses between cones and luminosity type horizontal cells (LHC), it was previously found in this laboratory that repetitive red flashes progressively strengthened the LHC's response to red flash, whereas weakened the LHC's response to green flash; repetitive green flash remarkably depressed the LHC's red response, but caused little changes in the cell's green response. However, the detailed mechanisms underlying these phenomena are not entirely clear. In the present study, based on an ion-channel model described mainly in the form of Hodgkin-Huxley equations, possible mechanisms of the short-term synaptic modification are investigated. The simulation results suggest that: (1) the auto enhancement effect might be induced by the Ca2+-dependent process on the post synaptic AMPA receptors, which could lead to changes of the ionic channel's properties; (2) the asymmetric response to red- and green-flashes and the mutual chromatic suppression effects might be attributed to the regulatory effects on the presynaptic glutamate release. PMID- 17115218 TI - Use of a neural mass model for the analysis of effective connectivity among cortical regions based on high resolution EEG recordings. AB - Assessment of brain connectivity among different brain areas during cognitive or motor tasks is a crucial problem in neuroscience today. Aim of this work is to use a neural mass model to assess the effect of various connectivity patterns in cortical electroencephalogram (EEG) power spectral density, and investigate the possibility to derive connectivity circuits from EEG data. To this end, a model of an individual region of interest (ROI) has been built as the parallel arrangement of three populations, each described as in Wendling et al. (Eur J Neurosci 15:1499-1508, 2002). Connectivity among ROIs includes three parameters, which specify the strength of connection in the different frequency bands. The following main steps have been followed: (1) we analyzed how the power spectral density (PSD) is significantly modified by the kind of coupling hypothesized among the ROIs; (2) with the model, and using an automatic fitting procedure, we looked for a simple connectivity circuit able to reproduce PSD of cortical EEG in three ROIs during a finger-movement task. The estimated parameters represent the strength of connections among the ROIs in the different frequency bands. Cortical EEGs were computed with an inverse propagation algorithm, starting from measurement performed with 96 electrodes on the scalp. The present study suggests that the model can be used as a simulation tool, able to mimic the effect of connectivity on EEG. Moreover, it can be used to look for simple connectivity circuits, able to explain the main features of observed cortical PSD. These results may open new prospectives in the use of neurophysiological models, instead of empirical models, to assess effective connectivity from neuroimaging information. PMID- 17115219 TI - Computer modelling of the sinoatrial node. AB - Over the past decades patch-clamp experiments have provided us with detailed information on the different types of ion channels that are present in the cardiac cell membrane. Sophisticated cardiac cell models based on these data can help us understand how the different types of ion channels act together to produce the cardiac action potential. In the field of biological pacemaker engineering, such models provide important instruments for the assessment of the functional implications of changes in density of specific ion channels aimed at producing stable pacemaker activity. In this review, an overview is given of the progress made in cardiac cell modelling, with particular emphasis on the development of sinoatrial (SA) nodal cell models. Also, attention is given to the increasing number of publicly available tools for non-experts in computer modelling to run cardiac cell models. PMID- 17115220 TI - Molecular mapping of two cultivar-specific avirulence genes in the rice blast fungus Magnaporthe grisea. AB - Rice blast, caused by the fungus Magnaporthe grisea, is a globally important disease of rice that causes annual yield losses. The segregation of genes controlling the virulence of M. grisea on rice was studied to establish the genetic basis of cultivar specificity in the interaction of rice and M. grisea. The segregation of avirulence and virulence was studied in 87 M. grisea F(1) progeny isolates from a cross of two isolates, Guy11 and JS153, using resistance gene-differential rice cultivars. The segregation ratio indicated that avirulence and virulence in the rice cultivars Aichi-asahi and K59, respectively, are controlled by single major genes. Genetic analyses of backcrosses and full-sib crosses in these populations were also performed. The chi(2 )test of goodness-of fitness for a 1:1 ratio indicated that one dominant gene controls avirulence in Aichi-asahi and K59 in this population. Based on the resistance reactions of rice differential lines harboring known resistance genes to the parental isolates, two genetically independent avirulence genes, AVR-Pit and AVR-Pia, were identified. Genetic linkage analysis showed that the SSR marker m355-356 is closely linked to AVR-Pit, on the telomere of chromosome 1 at a distance of approximately 2.3 cM. The RAPD marker S487, which was converted to a sequence-characterized amplified region (SCAR) marker, was found to be closely linked to AVR-Pia, on the chromosome 7 telomere at a distance of 3.5 cM. These molecular markers will facilitate the positional cloning of the two AVR genes, and can be applied to molecular-marker-assisted studies of M. grisea populations. PMID- 17115221 TI - T cells remaining after intensive chemotherapy for acute myelogenous leukemia show a broad cytokine release profile including high levels of interferon-gamma that can be further increased by a novel protein kinase C agonist PEP005. AB - Cytokines are released during T cell activation, including the potentially anti leukemic interferon-gamma (IFNgamma), but also the hematopoietic growth factor granulocyte-macrophage colony-stimulating factor (GM-CSF) that enhance proliferation and inhibit apoptosis of acute myelogenous leukemia (AML) cells. In the present study we investigated the release of IFNgamma and GM-CSF by circulating T cells in AML patients with chemotherapy-induced cytopenia. T cells were activated with anti-CD3 plus anti-CD28 in a whole-blood assay in the presence of their natural cytokine network. We examined 63 samples derived from 16 AML patients during 28 chemotherapy cycles. Activated T cells showed a broad cytokine release profile, but IFNgamma and GM-CSF levels showed a significant correlation and were generally higher than the other cytokine levels. Higher IFNgamma and GM-CSF responses were associated with a low CD4:CD8 ratio, older patient age and no ongoing chemotherapy indicating potential utility of T cell activation regimes for the older AML patient. The cytokine levels could be further increased by the novel protein kinase C agonist PEP005, which also induced significant production of IL2 and TNFalpha which could contribute to anti tumor effects in AML patients. We conclude that remaining T cells after intensive AML therapy show a broad cytokine release profile including high and significantly correlated levels of potentially anti-leukemic IFNgamma and the AML growth factor GM-CSF. The final outcome of an AML-initiated T cell cytokine response will thus depend on the functional characteristics of the AML cells, in particular the relative expression of IFNgamma and GM-CSF receptors which differs between AML patients. PMID- 17115222 TI - Objects do not aid inhibition of return in crossing the vertical meridian. AB - Location-based cuing experiments have shown that inhibition of return (IOR) spreads beyond a cued location but appears to be confined to the cued hemifield by the vertical meridian. Previous studies have also shown that IOR can spread across objects and here we investigate whether an object can be used to mediate the spreading of IOR into the opposite hemifield. Two experiments used a rectangular object that surrounded four target locations, two to the left and right of a central fixation point. The spreading of IOR in the presence of the object was determined and compared with a condition where the object-frame was absent. Object-present and object-absent trials were either mixed within a block (Experiment 1) or divided into separate blocks (Experiment 2). Both experiments revealed robust inhibition in the cued but not the uncued hemifield, further demonstrating the hemifield-based spreading of IOR. PMID- 17115223 TI - Habitat requirements of the seabird tick, Ixodes uriae (Acari: Ixodidae), from the Antarctic Peninsula in relation to water balance characteristics of eggs, nonfed and engorged stages. AB - The seabird tick Ixodes uriae is exposed to extreme environmental conditions during the off-host phase of its life cycle on the Antarctic Peninsula. To investigate how this tick resists desiccation, water requirements of each developmental stage were determined. Features of I. uriae water balance include a high percentage body water content, low dehydration tolerance limit, and a high water loss rate, which are characteristics that classify this tick as hydrophilic. Like other ticks, I. uriae relies on water vapor uptake as an unfed larva and enhanced water retention in the adult, while nymphs are intermediate and exploit both strategies. Stages that do not absorb water vapor, eggs, fed larvae and fed nymphs, rely on water conservation. Other noteworthy features include heat sensitivity that promotes water loss in eggs and unfed larvae, an inability to drink free water from droplets, and behavioral regulation of water loss by formation of clusters. We conclude that I. uriae is adapted for life in a moisture-rich environment, and this requirement is met by clustering in moist, hydrating, microhabitats under rocks and debris that contain moisture levels that are higher than the tick's critical equilibrium activity. PMID- 17115224 TI - FM signals produce robust paradoxical latency shifts in the bat's inferior colliculus. AB - Previous studies in echolocating bats, Myotis lucifugus, showed that paradoxical latency shift (PLS) is essential for neural computation of target range and that a number of neurons in the inferior colliculus (IC) exhibit unit-specific PLS (characterized by longer first-spike latency at higher sound levels) in response to tone pulses at the unit's best frequency. The present study investigated whether or not frequency-modulated (FM) pulses that mimic the bat's echolocation sonar signals were equally effective in eliciting PLS. For two-thirds of PLS neurons in the IC, both FM and tone pulses could elicit PLS, but only FM pulses consistently produced unit-specific PLS. For the remainder of PLS neurons, only FM pulses effectively elicited PLS; these cells showed either no PLS or no response, to tone pulses. PLS neurons generally showed more pronounced PLS in response to narrow-band FM (each sweeping 20 kHz in 2 ms) pulse that contained the unit's best frequency. In addition, almost all PLS neurons showed duration independent PLS to FM pulses, but the same units exhibited duration-dependent PLS to tone pulses. Taken together, when compared to tone pulses, FM stimuli can provide more reliable estimates of target range. PMID- 17115225 TI - Repression of anti-apoptotic genes via AP-1 as a mechanism of apoptosis induction in ventricular cardiomyocytes. AB - Nitric oxide (NO) is increased under several pathophysiological, mainly inflammatory processes in the heart and has been characterized as an inducer of apoptosis in cardiomyocytes. The transcription factor activating protein-1 (AP-1) has been identified as a mediator of NO-induced apoptosis. Genes that are regulated by AP-1 under apoptotic conditions have not been identified yet. Therefore, we performed a microarray analysis with subsequent real-time polymerase chain reaction (PCR) to identify genes regulated by AP-1 in NO-induced ventricular cardiomyocytes of rats and tested the functional role of these genes in apoptosis. Cardiomyocytes were transformed with AP-1 decoy oligonucleotides for inhibition of AP-1 activity. These, as well as non-transformed control cells, were stimulated with the NO donor (+/-)-S-nitroso-N-acetylpenicillamine (SNAP, 100 microM) for 2 h. Some of the genes with differential gene expression on microarrays were further analysed by real-time PCR. Genes that are induced by SNAP were not identified. However, four genes, pyridoxal kinase, heat shock protein 10 (Hsp10), antigen identified by monoclonal antibodies 4F2 (4F2) and myosin light chain 2, were downregulated by SNAP in presence of AP-1. Pyridoxal kinase, Hsp10 and 4F2 have anti-apoptotic effects in unstimulated cells because downregulation of their expression by antisense oligos induced apoptosis in cardiomyocytes. An involvement of these genes in NO-induced apoptosis could only be proven for pyridoxal kinase. In conclusion, using microarray technology, we identified three anti-apoptotic genes (Hsp10, 4F2 and pyridoxal kinase) in ventricular cardiomyocytes, which may help the cells to resist some apoptotic stimuli. The downregulation of these genes results in cardiomyocyte apoptosis. Prevention of their downregulation may protect cardiomyocytes against apoptotic stimuli, and this may be of therapeutic benefit. PMID- 17115226 TI - Radiofrequency surgery of the tongue base in the treatment of snoring--a pilot study. AB - In a previously published study, a significant reduction of snoring was reported after treatment with radiofrequency surgery of the tongue base in patients suffering from obstructive sleep apnea syndrome. The aim of this study was to investigate the efficacy of radiofrequency surgery of the tongue base in the treatment of primary snoring. Twenty patients suffering from primary snoring (AHI < 10/h, body mass index < 32 kg/m(2)) and an isolated hypertrophic tongue base at clinical examination were enrolled in this clinical trial. The patients underwent bipolar radiofrequency surgery of the tongue base under local anaesthesia. Pre- and post-operative body weight, daytime sleepiness (Epworth sleepiness scale) and snoring scores (visual analogue scales) were evaluated by the patients and their bed partners, respectively. Postoperative follow-up data was collected 6-8 weeks after treatment. A statistically significant reduction of the preoperative snoring levels from 7.5 +/- 2.4 to 6.1 +/- 2.8 was seen after treatment (p < 0.001). Body weight and daytime sleepiness remained unaffected. Only 3 out of 20 patients were satisfied with the result as defined by VAS < 3. Despite statistically significant reduction of the subjective snoring scores after radiofrequency of the tongue base, only minimal clinical improvement was achieved. Only 3 out of 20 patients were satisfied with the results. With regard to the clearly beneficial effect seen in patients with obstructive sleep apnea, this result indicates different pathophysiological principles in the generation of snoring. PMID- 17115227 TI - Pediatric arteriovenous malformation: University of Toronto experience using stereotactic radiosurgery. AB - INTRODUCTION AND BACKGROUND: Arteriovenous malformations (AVMs) are congenital vascular lesions of the brain, which behave differently in pediatric population compared to adults. Treatment of pediatric AVMs includes a combination of microsurgery, embolization and radiation therapies. However, the role of radiosurgery in the treatment of pediatric AVMs is not fully accepted because of concerns regarding the long-term effects of radiation on the pediatric brain. DISCUSSION: In this study, we review our experience at the University of Toronto with treating pediatric AVMs using linear accelerator-based (LINAC) radiosurgery over the past 15 years. We report our results, obliteration rates, and complications on a total of 40 patients. In addition, we provide a review of series published to date combined with our own results to determine whether radiosurgery is a safe and reasonable treatment modality for pediatric AVMs. PMID- 17115228 TI - Chondroblastoma of the apex portion of petrousal bone. AB - BACKGROUND: Skull chondroblastoma is extremely rare. We described in this study the first case of petrousal bone apex chondroblastoma. METHODS: The tumor occurred in a 12-year-old girl and was revealed by left hypoacusis, vertigo, and walking instability. Computerized tomography scan depicted well-demarcated and osteolytic lesion of the left petrousal bone just before the internal acoustic meatus. The tumor was operated on by a subtemporal approach with a subtotal resection. Lesion recurred and was operated a second time by a combined approach subtemporal and translabyrinthine. This treatment was completed by radiotherapy. We used a combination of photon therapy and proton therapy. Outcome was good and free of recurrence 36 months after the second operation. CONCLUSION: Proton therapy can be particularly useful for skull base tumors frequently radioresistant and near very important structures. This case is the first one of a successful treatment by proton therapy for chondroblastoma recurrence. PMID- 17115229 TI - Laparoscopic appendectomy is a favorable alternative for complicated appendicitis in children. AB - Laparoscopic appendectomy in children is a generally accepted procedure for the treatment of non-complicated acute appendicitis. Nevertheless, the role of laparoscopy in complicated appendicitis is more controversial. The objective of this study was to examine the safety, efficacy and complications of laparoscopy in children with complicated appendicitis. This is a retrospective review of the children who underwent laparoscopic appendectomy for complicated appendicitis at King Khalid University Hospital, Riyadh, Saudi Arabia between January 1998 and March 2006. Complicated appendicitis includes perforated appendicitis, gangrenous appendicitis and appendicular masses found intra-operatively. Data collected include demographic, duration of symptoms, operative time, analgesia, complications, length of hospitalization and histopathology. Laparoscopic appendectomy was done by three trocar technique in all cases. During the study period, 59 children aged 3-12 years underwent laparoscopic appendectomy for complicated appendicitis. There were 34 patients with perforated appendicitis, 12 patients with gangrenous appendicitis and 13 patients with appendicular mass. The average operating time was 62 min. The average length of hospitalization was 5 days. The post-operative narcotic analgesic requirement was minimal. Laparoscopy was converted to open surgery in two patients (3.38%). These two cases were excluded from further analysis. Four out of 57 patients (7.01%) had post operative complications. Three patients (5.26%) developed wound infection. One patient (1.75%) developed haematoma at umbilical port site. There was no post operative intra-abdominal collection. Laparoscopic appendectomy is a safe alternative for the treatment of complicated appendicitis. It does not increase the incidence of complications even with complicated appendicitis. Contrary to the previous studies, we did not have increased incidence of intra-abdominal collection in this review. However, prospective randomized controlled trials are needed to verify these findings. PMID- 17115230 TI - Neoadjuvant chemotherapy for bladder cancer. AB - The 30-45% failure rate after radical cystoprostatectomy mandates that we explore and optimize multimodal therapy to achieve better disease control in these patients. Cisplatin-based multi-agent combination chemotherapy has been used with success in metastatic disease and has therefore also been introduced in patients with high-risk but non-metastatic bladder cancer. There is now convincing evidence that chemotherapy given pre-operatively can improve survival in these patients. In this review we establish the need for peri-operative chemotherapy in bladder cancer patients and summarize the evidence for the efficacy of neoadjuvant chemotherapy. The advantages and disadvantages of neoadjuvant versus adjuvant chemotherapy are discussed, and the main shortcomings of both--treatment related toxicity and the inability to prospectively identify likely responders- are presented. Finally, a risk-adapted approach to neoadjuvant chemotherapy is presented, whereby the highest risk patients are offered treatment while those unlikely to benefit are spared the treatment-related toxicity. PMID- 17115231 TI - Sexual function following surgery for urodynamic stress incontinence. AB - The objective of this study was to compare sexual function in women before and after surgery for urodynamic stress incontinence in the absence of pelvic organ prolapse. This was a prospective questionnaire survey. Fifty-four women undergoing surgery (tension-free vaginal tape/tension-free vaginal tape obturator) for urodynamic stress incontinence with no evidence of detrusor overactivity or concomitant prolapse were assessed preoperatively and 6 months post operatively. Assessment was based on the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ), the International Consultation on Incontinence Questionnaire (ICIQ) and the Patient Global Impression of Improvement. Paired t-tests were used for comparing pre- and post-op scores and unpaired t-tests for comparing observations between groups. Spearman's rank correlation was used for testing whether two numerically scored items were related, and McNemar test was used to compare pre- and postoperative responses to individual questions. ICIQ scores showed significant improvement after surgery (p < 0.001). Women completing PISQ were significantly younger (mean = 54) than those who did not (mean = 65; p < 0.001). The total PISQ score was better postoperatively (preoperative = 87.2, postoperative = 92.7; p < 0.001), with improvements in both the physical (preoperative = 31.0, postoperative = 35.2; p < 0.001) and partner-related domains (preoperative = 18.8, postoperative = 19.9; p = 0.002) but no improvement in behaviour emotive domains (preoperative = 37.3, postoperative = 37.6; p = 0.70). There was a reduction in episodes of coital incontinence postoperatively (preoperatively = 16/54, postoperatively = 39/54; p < 0.002). Previous vaginal surgery, oestrogen status of respondents and hysterectomy status did not affect the PISQ. Surgical correction of stress incontinence is associated with an improvement in sexual function. PMID- 17115232 TI - Variability of current symptoms in women with pelvic organ prolapse. AB - To describe the variability of current (day-to-day) symptoms of pelvic organ prolapse (POP) and to compare women's overall (comprehensive) and current ratings of prolapse symptoms. This was a prospective observational study of 45 women with symptomatic POP followed-up for 3 months between November 2004 and June 2005. Women had a total of three visits during the study period. We measured current symptoms using daily symptom diaries completed for a total of 4 weeks during the study period. The diaries included 100-mm visual analogue scales anchored at 0 (no symptoms) and 100 (extremely severe symptoms) and patients completed these at three different times each day. At each study visit, patients were examined and completed a 10-item questionnaire, which included questions adapted from the PFDI. This questionnaire compared overall and current symptoms at each study visit. Statistical analyses included descriptive statistics, ANOVA, and Pearson's correlation coefficient. Current symptoms of vaginal bulging varied on a day-to day basis. Current symptoms varied depending on the time of day with women reporting more severe symptoms in the evenings (P<.01). Women reported experiencing their most severe symptoms at only 13.7% of all study visits. On average, women reported less severe ratings of current symptoms at the time of each visit compared to overall recall ratings (P<.05). Women seeking care for symptomatic POP can have dynamic symptoms and are often not evaluated at the time of their most severe symptoms. This should be considered by clinicians evaluating women for symptoms of POP. PMID- 17115233 TI - Accuracy and precision of a new portable ultrasound scanner, the BME-150A, in residual urine volume measurement: a comparison with the BladderScan BVI 3000. AB - The objective of the study was to determine the relative accuracy of a new portable ultrasound unit, BME-150A, and the BladderScan BVI 3000, as assessed in comparison with the catheterized residual urine volume. We used both of these machines to prospectively measure the residual urine volumes of 89 patients (40 men and 49 women) who were undergoing urodynamic studies. The ultrasound measurements were compared with the post-scan bladder volumes obtained by catheterization in the same patients. The ultrasounds were followed immediately (within 5 min) by in-and-out catheterizations while the patients were in a supine position. There were a total of 116 paired measurements made. The BME-150A and the BVI 3000 demonstrated a correlation with the residual volume of 0.92 and 0.94, and a mean difference from the true residual volume of 7.8 and 3.6 ml, respectively. Intraclass correlation coefficients for the accuracy of the two bladder scans were 0.90 for BME-150A and 0.95 for BVI 3000. The difference of accuracy between the two models was not significant (p = 0.2421). There were six cases in which a follow-up evaluation of falsely elevated post-void residual urine volume measurements on the ultrasound studies resulted in comparatively low catheterized volumes, with a range of differences from 66 to 275.5 ml. These cases were diagnosed with an ovarian cyst, uterine myoma, or uterine adenomyosis on pelvic ultrasonography. The accuracy of the BME-150A is comparable to that of the BVI 3000 in estimating the true residual urine volumes and is sufficient enough for us to recommend its use as an alternative to catheterization. PMID- 17115234 TI - Identification of novel RANK polymorphisms and their putative association with low BMD among postmenopausal women. AB - INTRODUCTION: Bone mineral density (BMD) is the major factor for determining bone strength, which is closely correlated to osteoporotic fracture risk and is largely determined by multiple genetic factors. The RANK (TNFRSF11A), receptor for RANKL, is a member of the tumor necrosis factor receptor (TNFR) superfamily and plays a central role in osteoclast development. METHODS: In order to investigate the effects of RANK polymorphism on BMD and osteoporosis, we directly sequenced the RANK gene in 24 Korean individuals and identified 25 sequence variants. Eleven of these polymorphisms were selected and genotyped in a larger scale study of postmenopausal women (n = 560). Areal BMD (g/cm(2)) of the anterior-posterior lumbar spine and the nondominant proximal femur were measured using dual-energy X-ray absorptiometry. RESULTS: We found that two intronic polymorphisms in the RANK gene [RANK + 34863G > A (rs12458117) and RANK + 35928insdelC (new polymorphism found in this study) in intron 6] were significantly associated with the BMD of the lumbar spine, i.e., rare alleles were significantly associated with low BMD of the lumbar spine among Korean postmenopausal women (p = 0.04 and 0.02, respectively). These polymorphisms were also associated with low BMD of proximal femur sites, including Ward's triangle, trochanter, and total femur. Our results suggest that +34863G > A and +35928insdelC polymorphisms in RANK are possible genetic factors for low BMD in postmenopausal women. PMID- 17115235 TI - In vivo confocal scanning laser microscopy in dermatology. AB - The need to improve the diagnostic accuracy and sensitivity for skin tumours has led to the development of new non-invasive, in vivo techniques including ultrasound, dermoscopy, digital photography, confocal scanning laser microscopy, magnetic resonance imaging and optical coherence tomography. Of all these in vivo techniques, only confocal scanning laser microscopy allows for the examination of the epidermis and papillary dermis at a resolution approaching histological detail. This review article looks at some of the most important applications of this new technology, highlighting its qualities and limitations. PMID- 17115236 TI - A comparative SEM study between hand instrument and Er:YAG laser scaling and root planing. AB - Scaling and root planing are one of the most commonly used procedures for the treatment of periodontal diseases. Removal of calculus using conventional hand instruments is incomplete and rather time-consuming. In search for more efficient and less difficult instrumentation, investigators have proposed lasers as alternatives or adjuncts for scaling and root planing. The aim of the present study was to compare the effectiveness of subgingival scaling and root planing with erbium: yttrium, aluminium, garnet (Er:YAG) laser and hand instrumentation in vitro. The mesial and distal surfaces of 15 periodontal loosed extracted teeth were treated randomly either by hand instrumentation or by Er:YAG laser irradiation. After choosing the "very long pulse mode" (pulse duration of about 700 micros), the output energy of 160 mJ with 920-microm beam diameter (RO7 Perio tip, Fidelis, Fotona, Slovenia) and frequency of 12 Hz were selected, both according to the best results of past studies. In addition, air water spray was used during the procedures. The morphology of the root surface was evaluated by three observers with a scanning electron microscopy in magnifications of 50x and 400x. The result of this setting showed that the rate of remained roughness on treated root surfaces in two groups of hand instruments and Er:YAG laser had a meaningful difference: The surface roughness in Er:YAG laser group was more than in hand instruments group. The present study could demonstrate the in vitro capability of the Er:YAG laser for scaling and root planing in periodontitis, although the effectiveness of this setting did not reach that achieved by hand instrumentation. It could be concluded that lower frequency and long pulse duration maybe more suitable for the micro-morphology of root surface after treatment. This theory is going to be tested with the same laser instrument in the next study. PMID- 17115237 TI - Comparative evaluation of the effects of Nd:YAG and Er:YAG laser in dentin hypersensitivity treatment. AB - Dentin hypersensitivity (DH) is one of the most common complications that affect patients after periodontal therapy. So far, many investigators have successfully used different types of laser on DH treatment. The aim of this study was to evaluate the comparative effect of Nd:YAG laser and Er:YAG laser on human teeth desensitization. A group of nine patients with a total of 63 chronic hypersensitive teeth were selected. Each one of them should at least have three hypersensitive teeth. These teeth were randomly allocated into three groups. Group 1, Nd:YAG laser (1 W, 15 Hz, 60 s, two times); group 2, Er:YAG laser (100 mJ, 3 Hz, 60 s, two times); and group 3 serves as control group without any treatment. Assessment of pain was performed by a visual analysing scale (VAS) after stimulation of sensitive tooth by using the sharp tip of an explorer. This test was performed before treatment, immediately after that and at 1-, 3- and 6 month intervals after treatment by one blinded examiner. Analysis of VAS score between the three groups at the time of treatment did not show any significant difference (p = 0.506). However, by using repeated-measurement analysis of variance test, significant differences were seen in the three groups between before-treatment VAS score and after treatment (p < 0.0005). This statistically significant difference in the control group demonstrated a placebo effect. However, the effect of using Nd:YAG and Er:YAG lasers was stronger than this placebo effect, so that after removing the effect of the placebo, differences immediately after, 1, 3 and 6 months post treatment between all three groups still were statistically highly significant (p < 0.0005). Compared to the Er:YAG laser group, using Nd:YAG laser resulted in a significant reduction of VAS score at each follow-up examination (p < 0.0005). Although using Nd:YAG and Er:YAG laser in desensitization of hypersensitive teeth showed a placebo effect limited to a short time, results of this study demonstrated that both of these lasers have an acceptable therapeutic effect. The observed effects seemed to last for at least 6 months. It was concluded that Nd:YAG laser is more effective than Er:YAG laser in reduction of patients' pain. PMID- 17115238 TI - Effect of laser soldering irradiation on covalent bonds of pure collagen. AB - Laser tissue welding and soldering is being increasingly used in the clinical setting for defined surgical procedures. The exact induced changes responsible for tensile strength are not yet fully investigated. To further improve the strength of the bonding, a better understanding of the laser impact at the subcellular level is necessary. The goal of this study was to analyze whether the effect of laser irradiation on covalent bonding in pure collagen using irradiances typically applied for tissue soldering. Pure rabbit and equine type I collagen were subjected to laser irradiation. In the first part of the study, rabbit and equine collagen were compared using identical laser and irradiation settings. In the second part of the study, equine collagen was irradiated at increasing laser powers. Changes in covalent bonding were studied indirectly using the sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) technique. Tensile strengths of soldered membranes were measured with a calibrated tensile force gauge. In the first experiment, no differences between the species-specific collagen bands were noted, and no changes in banding were found on SDS-PAGE after laser irradiation. In the second experiment, increasing laser irradiation power showed no effect on collagen banding in SDS-PAGE. Finally, the laser tissue soldering of pure collagen membranes showed virtually no determinable tensile strength. Laser irradiation of pure collagen at typical power settings and exposure times generally used in laser tissue soldering does not induce covalent bonding between collagen molecules. This is true for both rabbit and equine collagen proveniences. Furthermore, soldering of pure collagen membranes without additional cellular components does not achieve the typical tensile strength reported in native, cell-rich tissues. This study is a first step in a better understanding of laser impact at the molecular level and might prove useful in engineering of combined collagen-soldering matrix membranes for special laser soldering applications. PMID- 17115239 TI - MTHFR polymorphism and bone mineral density: meta-analysis of published studies. AB - The C677T (rs1801133) polymorphism of methylenetetrahydrofolate reductase (MTHFR) has been associated with bone status in some studies, but the results have been mixed. In order to have a better understanding of this issue, we performed a meta analysis of studies about the association of the C677T polymorphism and bone mineral density (BMD). Eight studies analyzed the relationship with spine BMD. When their results were combined, individuals with TT genotype showed a small but significantly reduced BMD compared to those with TC and CC genotypes. The weighted mean difference (WMD) was 18.0 mg/cm2 (P = 0.001, 95% confidence interval [CI] 7.1-28.9), without statistical evidence for between-study heterogeneity (P = 0.28, I2 = 17%). Six studies analyzed femoral neck BMD. A test for heterogeneity was significant (P = 0.03, I2 = 56%). Individuals with TT alleles tended to have somewhat lower BMD, but the difference was not statistically significant. In random effects model, the WMD between the TT and TC/CC genotypes was 6.4 mg/cm2 (95% CI -7.8 to 21.2, P = 0.36). Total hip BMD was measured in four studies. They showed a significantly lower BMD in subjects with TT alleles: WMD 19.7 (95% CI 5.3-34.1) mg/cm2, P = 0.007, in comparison with TC/CC subjects. When we considered only studies on women, the WMD in BMD between TT and TC/CC genotypes was significant at the spine (22.1 mg/cm2, 95% CI 8.6 35.6; P = 0.001) and the femoral neck (15.5 mg/cm2, 95% CI 4.3-26.7; P = 0.007). There was no evidence for heterogeneity. The small number of studies did not allow a meaningful sex-stratified analysis of total hip BMD or a separate analysis of male data. In conclusion, the C677T polymorphism of the MTHFR gene is associated with small differences in BMD, at least in women. PMID- 17115240 TI - Racial differences in the association of subcutaneous and visceral fat on bone mineral content in prepubertal children. AB - Total fat mass plays a significant role in determining bone mass, but the specific role of central adiposity independent of total fat mass has not been widely studied. Prepubertal (Tanner 1) children (n = 181; 65 boys, 116 girls, 7.8 +/- 1.5 years), including 99 Caucasians and 82 African Americans from Birmingham, Alabama, participated in this study. Body composition, including total body and trunk fat mass, and bone mineral content (BMC) were measured using dual-energy X ray absorptiometry. Subcutaneous abdominal adipose tissue (SAAT) and intra abdominal adipose tissue (IAAT) were determined by single-slice computed tomography (CT). After adjusting for gender, age, height, total fat, and lean mass, trunk weight was inversely correlated with BMC in Caucasians (r = -0.56, P < 0.0001) and in African Americans (r = -0.37, P < 0.05). In Caucasians, independent of gender, age, height, total fat, and lean mass, there was an inverse correlation between SAAT and BMC (r = -0.58, P < 0.0001) but no significant correlation between IAAT and BMC; in addition, SAAT explained 6% of the variance in BMC. In contrast, in African Americans, SAAT and BMC were not significantly correlated. However, while adjusting for gender, age, height, SAAT, total fat, and lean mass, an inverse association between IAAT and BMC was observed in African Americans (r = -0.50, P < 0.01); IAAT also explained 3% of the variance in BMC. These findings suggest that, in general, total abdominal weight is negatively associated with bone mass, but there appear to be racial differences with regard to the contributions of subcutaneous and visceral fat to BMC in prepubertal children. PMID- 17115241 TI - Mechanism by which MLO-A5 late osteoblasts/early osteocytes mineralize in culture: similarities with mineralization of lamellar bone. AB - The mechanisms whereby bone mineralizes are unclear. To study this process, we used a cell line, MLO-A5, which has highly elevated expression of markers of the late osteoblast such as alkaline phosphatase, bone sialoprotein, parathyroid hormone type 1 receptor, and osteocalcin and will mineralize in sheets, not nodules. In culture, markers of osteocytes and dendricity increase with time, features of differentiation from a late osteoblast to an early osteocyte. Mineral formation was examined using transmission electron microscopy, scanning electron microscopy with energy-dispersive X-ray analysis, and atomic force microscopy. At 3-4 days of culture, spheres of approximately 20-50 nm containing calcium and phosphorus were observed budding from and associated with developing cellular projections. By 5-6 days, these calcified spheres were associated with collagen fibrils, where over time they continued to enlarge and to engulf the collagen network. Coalescence of these mineralized spheres and collagen-mediated mineralization were responsible for the mineralization of the matrix. Similar calcified spheres were observed in cultured fetal rat calvarial cells and in murine lamellar bone. We propose that osteoid-osteocytes generate spherical structures that calcify during the budding process and are fully mineralized on their developing cellular processes. As the cellular process narrows in diameter, these mineralized structures become associated with and initiate collagen mediated mineralization. PMID- 17115243 TI - Urethral injury as a complication of isolated diastasis pubis: case report. AB - Diastasis of the symphysis pubis is typically associated with straddle-type pelvic trauma. A case of diastasis pubis due to a recreational water-ski accident is described here. This case is complicated with a partial urethral rupture. PMID- 17115244 TI - Case report: primary carcinoid tumor of the testicle without metastases in combination with testicular atrophy and testosterone deficiency. AB - The first case of testicular carcinoid was represented as an element of a benign cystic teratoma by Simon et al. J Urol 1954; 72: 892-894. It is a rare disease accounting for less than 1% of all testicular neoplasms. We report a case of carcinoid of the testis without carcinoid syndrome and metastasis but with testosterone deficiency based on a bilateral testicular atrophy, which has not been previously reported. PMID- 17115245 TI - A large seminal vesicle cyst with contralateral renal agenesis. AB - Seminal vesicle cysts (SVC) are rather rare disorders. Our case is the first in literature where contralateral renal agenesis was seen together with SVC and surgically managed. We believe that the occurrence of these two coinciding abnormalities is incidental. PMID- 17115246 TI - Solitary hydatid cyst in the pelvis: a case report. AB - Hydatid disease mainly affects the liver and the lungs. Pelvic involvement have been rarely reported in the literature. Herein we present a rare case of isolated hydatid cyst of pelvis attached to the urinary bladder. PMID- 17115247 TI - Treatment of bladder cancer in cardiac transplant patients. AB - We report a patient who presented with superficial bladder tumors 8 years after cardiac transplantation. He was managed conservatively with intra-vesical therapy for 5 years and ultimately underwent a successful radical cystoprostatectomy and urethrectomy 13 years after cardiac transplantation. This illustrates that cardiac transplant patients with superficial tumors may be managed with conservative therapies, but close vigilance is necessary to identify the opportune time for extirpative surgery. PMID- 17115248 TI - Laparoscopic nephroplication and nephropexy as an adjunct to pyeloplasty in UPJO with giant hydronephrosis. AB - Giant hydronephrosis due to uretero-pelvic junction obstruction (UPJO) is rare and usually the treatment is nephrectomy. When renal salvage is required, reconstruction of the UPJ alone is usually inadequate to improve drainage from the pelvicalyceal system. Various open surgical techniques have been described to improve gravity-assisted drainage from the giant hydronephrotic kidney. We describe, for the first time, a complete laparoscopic approach for pyeloplasty combined with nephroplication and nephropexy. PMID- 17115249 TI - Radiotherapy for rectal carcinoma: an unusual cause of foreskin phimosis. AB - Phimosis of the foreskin secondary to radiotherapy for a pelvic malignancy has not been previously described in the world literature. However, as radiotherapy is ever more widely used in the treatment of various pelvic malignancies, it is important to ensure that this complication is prevented by the use of appropriate penile shielding. PMID- 17115250 TI - Molecular cloning of Brassica napus TRANSPARENT TESTA 2 gene family encoding potential MYB regulatory proteins of proanthocyanidin biosynthesis. AB - Three members of Brassica napus TRANSPARENT TESTA 2 (BnTT2) gene family encoding potential R2R3-MYB regulatory proteins of proanthocyanidin biosynthesis were isolated. BnTT2-1, BnTT2-2, and BnTT2-3 are 1102 bp with two introns, and have a 938-bp full-length cDNA with a 260 amino acid open reading frame. They share 98.2 99.3% nucleotide and 96.5-98.5% amino acid identities to each other, and are orthologous to Arabidopsis thaliana TT2 (AtTT2) with 74.1-74.8% nucleotide and 71.1-71.8% amino acid identities. An mRNA type of BnTT2-2 was found to contain unspliced intron 2 and encode a premature protein. They all have an alternative polyadenylation site. BnTT2-1 and BnTT2-3 also have an alternative transcription initiation site. Aligned with AtTT2, their 5' untranslated regions (UTRs) are astonishingly conserved, and two conserved regions were also found in their 3' UTRs. Oligonucleotide deletion leads to double-start codons of them. Resembling AtTT2, BnTT2 proteins are nuclear-located R2R3-MYB proteins containing predicted DNA-binding sites, bHLH interaction residues, and transcription activation domains. Southern blot indicated that there might be three BnTT2 members in B. napus, lower than triplication-based prediction. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) revealed that the expression of BnTT2-2 is mostly like AtTT2 with intensive expression in young seeds, but it is also expressed in root in which AtTT2 has no expression. BnTT2-1 shows lower tissue specificity and transcription levels, whereas BnTT2-3 is the lowest. Comparative cloning and RT-PCR indicated that seed color near-isogenic lines L1 and L2 have equivalent BnTT2 genes, and the yellow seed color in L2 might be caused by locus/loci other than BnTT2. Our results lay the basis for further investigating the regulatory mechanism of BnTT2 genes in flavonoid pathway and for transgenic creation of novel yellow-seeded B. napus stocks. PMID- 17115251 TI - Group I intron renders differential susceptibility of Candida albicans to Bleomycin. AB - The alarming increase in drug resistance gained by fungal pathogens has raised an urgent need to develop drugs against novel targets. Candida albicans, an opportunistic fungal pathogen, harbors in its 25S rRNA gene, a self-splicing Group I intron, which can act as a selective drug target. We report that Bleomycin selectively inhibits the self-splicing of Group I intron of C. albicans at IC(50) = 1.2 microM, leading to accumulation of precursor RNA as evinced by Reverse Transcriptase PCR. Drug susceptibility assays including MIC determination, growth curve analysis and disc diffusion assays indicate a strong susceptibility of the intron-containing strain (4-1) than the intronless strain (62-1). These results on the preferential targeting of Group I intron of C. albicans by Bleomycin might form a basis for design of small molecules that inhibit self-splicing of RNA as a antimicrobial tool against life-threatening microorganisms. PMID- 17115252 TI - Attitudes of medical students towards psychiatry : effects of training, courses in psychiatry, psychiatric experience and gender. AB - OBJECTIVE: The attitudes of medical students towards psychiatry and psychotherapy were examined considering the extent of their education, previous psychiatry experience, the evaluation of the course, their career intentions and socio demographic variables. METHODS: Five hundred and eight medical students in their second, fifth, ninth and tenth semester completed a questionnaire on "Attitudes Towards Psychiatry" (ATP-30). RESULTS: With 508 participants (return quota: 88%), the study showed no overall change in attitude in the ATP-30 during the course of the study. No general change in attitude following practical training in psychiatry was found. The more positive the psychiatry course was rated the more positive attitudes towards psychiatry were. Female students and students with psychiatry/psychotherapy experience had a significantly more positive attitude towards psychiatry as a subject. Overall 5.8% of all students showed a very positive attitude towards psychiatry. CONCLUSION: A positive education experience as well as personal experience increases the probability of a positive student attitude towards psychiatry. In order to assure adequate care for the mentally ill, it should be a fundamental aim of medical education to promote positive attitudes towards the mentally ill and psychiatry. It remains to be investigated, however, whether an improvement in the attitudes of students towards the psychiatry discipline is sufficient to increase the number of students who would like to become psychiatrists or whether other factors are more deciding such as career opportunities, conditions of further education, or income potential. PMID- 17115253 TI - A study of crop-to-crop gene flow using farm scale sites of fodder maize (Zea mays L.) in the UK. AB - From 2000 to 2003 a range of Farm Scale Evaluation (FSE) trials were established in the UK to assess the effect of the release and management of herbicide tolerant (HT) crops on arable weeds and invertebrates. The FSE trials for maize were also used to investigate crop-to-crop gene flow and to develop a statistical model for the prediction of gene flow frequency that can be used to evaluate current separation distance guidelines for GM crops. Seed samples were collected from the non-GM half of 55 trial sites and 1,055 were tested for evidence of gene flow from the GM HT halves using a quantitative PCR assay specific to the HT (pat) gene. Rates of gene flow were found to decrease rapidly with increasing distance from the GM source. Gene flow was detected in 30% of the samples (40 out of 135) at 150 m from the GM source and events of GM to non-GM gene flow were detected at distances up to and including 200 m from the GM source. The quantitative data were subjected to statistical analysis and a two-step model was found to provide the best fit for the data. A dynamic whole field model predicted that a square field (150 m x 150 m in size) of grain maize would require a separation distance of 3 m for the adjacent crop to be below a 0.9% threshold (with <2% probability of exceeding the threshold). The data and models presented here are discussed in the context of necessary separation distances to achieve various possible thresholds for adventitious presence of GM in maize. PMID- 17115254 TI - Sequence representation and prediction of protein secondary structure for structural motifs in twilight zone proteins. AB - Characterizing and classifying regularities in protein structure is an important element in uncovering the mechanisms that regulate protein structure, function and evolution. Recent research concentrates on analysis of structural motifs that can be used to describe larger, fold-sized structures based on homologous primary sequences. At the same time, accuracy of secondary protein structure prediction based on multiple sequence alignment drops significantly when low homology (twilight zone) sequences are considered. To this end, this paper addresses a problem of providing an alternative sequences representation that would improve ability to distinguish secondary structures for the twilight zone sequences without using alignment. We consider a novel classification problem, in which, structural motifs, referred to as structural fragments (SFs) are defined as uniform strand, helix and coil fragments. Classification of SFs allows to design novel sequence representations, and to investigate which other factors and prediction algorithms may result in the improved discrimination. Comprehensive experimental results show that statistically significant improvement in classification accuracy can be achieved by: (1) improving sequence representations, and (2) removing possible noise on the terminal residues in the SFs. Combining these two approaches reduces the error rate on average by 15% when compared to classification using standard representation and noisy information on the terminal residues, bringing the classification accuracy to over 70%. Finally, we show that certain prediction algorithms, such as neural networks and boosted decision trees, are superior to other algorithms. PMID- 17115255 TI - Botulinum neurotoxin light chain refolds at endosomal pH for its translocation. AB - Botulinum neurotoxins (BoNTs), the most poisonous member of class A biothreat agent, cause neuroparalysis by blocking neurotransmitter release at the neuromuscular junctions. In its mechanism of action, the catalytic domain (light chain (LC) of BoNT) is transported to the cytosol by the heavy chain (HC) in order to reach its proteolytic substrates. The BoNT HC forms a membrane channel under acidic conditions encountered in endosomes to serve as a passageway for LC to enter into cytosol. We demonstrate here that BoNT/A LC undergoes unique structural changes under the low pH conditions, and adopts a molten globule state, exposing substantial number of hydrophobic groups. The flexibility of the molten globular structure combined with retention of the secondary structure and exposure of specific residues of LC for interaction with the HC, allows its translocation through the narrow endosomal membrane channel. PMID- 17115256 TI - Cloning and structural analysis of a haemocyanin from the Stonefly Perla grandis. AB - We characterized two subunits of a putative haemocyanin from the stonefly species Perla grandis. In particular, we cloned and sequenced the corresponding cDNAs and studied their expression in different insect stages. Moreover, using the deduced amino acid sequences, homology studies were performed both on their primary and tertiary structures. 3-D molecular modelling data showed that the residues involved in the oxygen transport and subunits contacts were located in spatial positions preserving the functionality of the molecule. Despite it was paradigmatically affirmed that insects do not have respiratory proteins, our data suggest that the haemocyanin could be involved in the respiratory mechanisms of P. grandis. As far as we know, this is the first haemocyanin 3-D structure described and analyzed in insects. PMID- 17115257 TI - Data-driven smooth tests of the proportional hazards assumption. AB - A new test of the proportional hazards assumption in the Cox model is proposed. The idea is based on Neyman's smooth tests. The Cox model with proportional hazards (i.e. time-constant covariate effects) is embedded in a model with a smoothly time-varying covariate effect that is expressed as a combination of some basis functions (e.g., Legendre polynomials, cosines). Then the smooth test is the score test for significance of these artificial covariates. Furthermore, we apply a modification of Schwarz's selection rule to choosing the dimension of the smooth model (the number of the basis functions). The score test is then used in the selected model. In a simulation study, we compare the proposed tests with standard tests based on the score process. PMID- 17115258 TI - Asymptotic theory for the Cox semi-Markov illness-death model. AB - Irreversible illness-death models are used to model disease processes and in cancer studies to model disease recovery. In most applications, a Markov model is assumed for the multistate model. When there are covariates, a Cox (1972, J Roy Stat Soc Ser B 34:187-220) model is used to model the effect of covariates on each transition intensity. Andersen et al. (2000, Stat Med 19:587-599) proposed a Cox semi-Markov model for this problem. In this paper, we study the large sample theory for that model and provide the asymptotic variances of various probabilities of interest. A Monte Carlo study is conducted to investigate the robustness and efficiency of Markov/Semi-Markov estimators. A real data example from the PROVA (1991, Hepatology 14:1016-1024) trial is used to illustrate the theory. PMID- 17115260 TI - Metal binding of metallothioneins in human astrocytomas (U87 MG, IPDDC-2A). AB - Astroglia cells structurally and nutritionally support neurons in the central nervous system. They play an important role in guiding the construction of the nervous system and controlling the chemical and ionic environment of neurons. They also represent the major sites for accumulation and immobilisation of toxic metal ions most probably connected with metallothioneins. For this reason astroglia cells possess high cytosolic levels of metallothioneins I, II and III (MT-I,II,III). Our aim was to establish the inducibility and metal binding of MTs in two human astrocytoma cell lines, U87 MG (astrocytoma-glioblastoma, grade IV) and IPDDC-2A (astrocytoma, grade II), on exposure to cadmium chloride (1 microM). MTs were identified by molecular weight (size exclusion chromatography) and their metal content (Cd, Zn and Cu) to follow the interactions between metals. We showed that MTs are constitutively expressed in both human astrocytoma cell lines. In accordance with the higher malignancy grade of U87 MG, the amount of MTs was higher in U87 MG than in IPDDC-2A cells. After 24 hours of exposure to Cd their expression greatly increased in both cell lines and they were capable of immobilising almost all water soluble Cd. Induction of MTs in U87 MG cells was additionally followed up to 48 hours with exposure to different concentrations of CdCl(2) (1, 10 microM). Induction was a time dependent process throughout the period. Isoform III (identified by chromatographic separation of isoform III from I/II) was present at all exposure times, but only in traces with respect to the prevailing amounts of MT-I/II isoforms. So induction can be attributed to isoform I/II only. PMID- 17115261 TI - Characteristics of copper tolerance in Yarrowia lipolytica. AB - We discovered that a mutant strain of the dimorphic yeast Yarrowia lipolytica could grow in the yeast form in high concentrations of copper sulfate. The amount of metal accumulated by Y. lipolytica increased with increasing copper concentrations in the medium. Washing with 100 mM EDTA released at least 60% of the total metal from the cells, but about 20-25 micromol/g DW persisted, which represented about 30% of the soluble fraction of cultured cells. The soluble fraction (mainly cytosol) contained only about 10% of the total metal content within cells cultured in medium supplemented with 6 mM copper. We suggest that although a high copper concentration induces an efflux mechanism, the released copper becomes entrapped in the periplasm and in other parts of the cell wall. Washing with EDTA liberated not only copper ions, but also melanin, a brown pigment that can bind metal and which located at the cell wall. These findings indicated that melanin participates in the mechanism of metal accumulation. Culture in medium supplemented with copper obviously enhanced the activities of Cu, Zn-SOD, but not of Mn-SOD. PMID- 17115262 TI - A comprehensive survey of brain interface technology designs. AB - In this work we present the first comprehensive survey of Brain Interface (BI) technology designs published prior to January 2006. Detailed results from this survey, which was based on the Brain Interface Design Framework proposed by Mason and Birch, are presented and discussed to address the following research questions: (1) which BI technologies are directly comparable, (2) what technology designs exist, (3) which application areas (users, activities and environments) have been targeted in these designs, (4) which design approaches have received little or no research and are possible opportunities for new technology, and (5) how well are designs reported. The results of this work demonstrate that meta analysis of high-level BI design attributes is possible and informative. The survey also produced a valuable, historical cross-reference where BI technology designers can identify what types of technology have been proposed and by whom. PMID- 17115263 TI - Illuminating the center: mechanisms regulating lumen formation and maintenance in mammary morphogenesis. AB - The lumens present in ductal structures are required for transport of fluids and air. Studies in model organisms and cells in culture suggest that lumens can be generated by multiple mechanisms including apoptosis of centrally located cells, and re-modeling of epithelia. Several studies point to a role for apoptosis during lumen formation in the mammary ducts. However, a role for other mechanisms during lumen formation in the mammary ducts is largely unexplored. Understanding how lumens are formed and maintained free of cells is of clinical importance because filling of the luminal space is associated with cancer and inflammation. Thus, further investigation can lead to new diagnostic and therapeutic opportunities. PMID- 17115264 TI - Mammary gland macrophages: pleiotropic functions in mammary development. AB - Mammary gland development is a complex process involving epithelial cells and supporting stromal cells. Macrophages (MOs) are an important component of the mammary gland stroma and are critical for normal mammary gland development; however, the mechanisms by which macrophages regulate these processes are not well understood. MOs are known to interact with numerous cell types, including epithelial cells, fibroblasts, adipocytes, and endothelial cells, all of which are significant components of mammary gland development. Therefore, the purpose of this review is to describe the interactions between MOs and these various cell types and use this knowledge to identify potential functions of MOs in the mammary gland. PMID- 17115265 TI - D-beta-hydroxybutyrate prevents glutamate-mediated lipoperoxidation and neuronal damage elicited during glycolysis inhibition in vivo. AB - Excitotoxic neuronal death mediated by over-activation of glutamate receptors has been implicated in ischemia, hypoglycemia and some neurodegenerative diseases. It involves oxidative stress and is highly facilitated during impairment of energy metabolism. We have shown previously that in vivo systemic glycolysis inhibition with iodoacetate (IOA), exacerbates glutamate excitotoxicity. We have now investigated whether this effect involves oxidative damage to membrane lipids, as evaluated by the presence of thiobarbituric acid-reactive substances. We have also tested whether the ketone body, D-beta-hydroxybutyrate (D-BHB), prevents lipoperoxidation and tissue damage. Results show that glutamate intrastriatal injection in control rats transiently enhances lipoperoxidation, while in IOA treated animals increased lipoperoxidation is sustained. Treatment with D-BHB significantly reduces striatal lesions and lipoperoxidation. Vitamin E also reduced neuronal damage and lipoperoxidation. Results suggest that glycolysis impairment favors a pro-oxidant condition and situates oxidative damage as an important mediator of in vivo induced excitotoxicity. Results provide evidence for the neuroprotective effect of D-BHB against glutamate toxicity. PMID- 17115266 TI - Ablation of difficult right-sided accessory pathways aided by mapping of tricuspid annular activation using a Halo catheter : Halo-mapping of right sided accessory pathways. AB - OBJECTIVE: To demonstrate that the use of a 20-pole catheter (Halotrade mark) positioned around the tricuspid valve annulus (TVA) is helpful in rapidly localising right free wall accessory pathways (AP), enhancing catheter stability during ablation, and leading to increased success in ablating these challenging pathways. PATIENTS AND METHODS: Seven consecutive patients who underwent Halo mapping of right-sided AP were studied. All but one had previously failed ablation. With a Halo catheter deployed at TVA, the accessory pathway location was rapidly identified using the sites of earliest atrial (A) activation during ventricular (V) pacing or orthodromic tachycardia, or earliest V-activation during sinus rhythm or A-pacing were identified. The stability of the ablation catheter was guided fluoroscopically (with reference to the stationary Halo), and electrically (contact artefact between the ablation catheter and Halo poles). RESULTS: AP locations were identified by the Halo (anterior in one patient, antero-lateral in one, lateral in two, and postero-lateral in three) where similar local VA/AV intervals were recorded at both the ablation catheter and Halo bipoles recording the shortest VA/AV intervals (four of seven patients), contact artefact between the ablation catheter and those Halo bipoles was seen (six of seven patients), or both (three of seven patients). All APs were ablated successfully after a mean RF duration of 5+/-2 min, and 25+/-17 min post Halo deployment without clinical recurrence at 12+/-4 months follow-up. CONCLUSION: A Halo positioned at the TVA can ease the localisation of right-sided AP, facilitate catheter stability during ablation, and guides successful ablation. PMID- 17115267 TI - Permanent direct his bundle pacing does not induce ventricular dyssynchrony unlike conventional right ventricular apical pacing. An intrapatient acute comparison study. AB - BACKGROUND: Benefits of A-V synchrony during right ventricular apical pacing are neutralized by induction of ventricular dyssynchrony. Only a few data are reported about direct His bundle pacing influence on ventricular synchronism. AIM: Was to assess the capability of direct His bundle pacing to prevent pacing induced ventricular dyssynchrony comparing DDD- (or VVI- in case of Atrial Fibrillation) right ventricular apical pacing with DDD- (or VVI-) direct His bundle pacing in the same patients cohort. METHODS: 23 of 24 patients (mean age 75.1 +/- 6.4 years) with narrow QRS (HV < 65 ms) underwent permanent direct His bundle pacing for "brady-tachy syndrome" (11) or supra-Hisian II/III-degree AV Block (permanent atrial fibrillation 7, AV Node ablation 1). A 4.1 F screw-in lead was fixed in His position, guided by endocardial pacemapping and unipolar recordings. Additional permanent (13 patients) or temporary right ventricular apical pacing leads were also positioned. Inter- and left intra-ventricular dyssynchrony, mitral regurgitation and left systolic ventricular function Tei index were assessed during either direct His bundle pacing or right ventricular apical pacing. RESULTS: Permanent direct His bundle pacing was obtained in 23 of 24 patients. Indexes of ventricular dyssynchrony were drastically reduced, mitral regurgitation decreased and left systolic ventricular function Tei index improved during direct His bundle pacing (or His bundle and septum pacing) in comparison to apical pacing (p < 0.05). No statistically significant differences were observed between direct His bundle pacing and combined His bundle and septum pacing. CONCLUSION: Direct His bundle pacing (also fused with adjacent septum capture) prevents pacing-induced ventricular dyssynchrony. PMID- 17115268 TI - Osteoclastic differentiation and function regulated by old and new pathways. AB - The osteoclast is a specialized multinucleated variant of the macrophage family. It degrades mineralized tissue, and is required for modeling and remodeling of bone. The osteoclast has long been known to require vitamin D for its differentiation and to be regulated by parathyroid hormone via circulating Ca(2+) levels. Two local signals important in osteoclast survival and differentiation, CSF-1 and RANKL, were characterized by the mid-1990 s. A basic framework of specialized cell attachment and resorption molecules was also clear by that time, including the alpha(v)beta(3) integrin, the key adhesion molecule of the mature osteoclast, the highly expressed vacuolar-type H(+)-ATPase that drives acid secretion to dissolve mineral, and cathepsin K, the predominant acid proteinase for collagenolysis. Recently, additional detail has been added to this framework, showing that the osteoclast has more complex regulation than was previously believed. These include the findings that one component of the V-H(+)-ATPase is unique to the osteoclast, that chloride transport and probably Cl(-)/H(+) exchange are also required for mineral degradation, and that additional receptors besides RANK and Fms regulate osteoclast formation and survival. Additional receptors include estrogen receptor-alpha, TNF-family receptors other than RANK, and, at least in some cases, glycoprotein hormone receptors including the TSH-R and the FSH-R. Challenges in understanding osteoclast biology include how the signalling mechanisms function cooperatively. Recent findings suggest that there is a network of cytoplasmic adapters, including Gab-2 and BCAR1, which are modified by multiple signalling mechanisms and which serve to integrate the signalling pathways. PMID- 17115269 TI - A "two-step" educational approach for patients taking oral anticoagulants does not improve therapy control. AB - BACKGROUND AND OBJECTIVE: The educational aspect of oral anticoagulant treatment is considered a possible cause of variability in anticoagulation levels. The aim of this prospective study was to investigate whether the time spent in the therapeutic range (TSTR) by patients taking oral anticoagulants could be improved by two different, consecutive educational approaches on the crucial aspects of oral anticoagulant therapy. DESIGN AND METHODS: Between May and June 2004, validated interviews were conducted with 240 patients (128 male and 112 female, mean age 50 +/- 12 years) enrolled in the study. Three months later, the patients were randomly allocated to three groups. A course that focused on the questions in the interview was followed by the first group (n = 80); a brochure containing the correct answers to questions was given to the second (n = 81); nothing was provided for the third (n = 79). RESULTS: A significant difference was found in the TSTR between the quarters preceding and following the interview. Mean TSTR increase was 13%. Patients that were randomly selected to attend the educational course, read a brochure or do nothing showed similar TSTR percentages in the quarter following the interview. A good control of the anticoagulant therapy (TSTR >70%) was maintained, with no significant variation during the following three-quarters. INTERPRETATION AND CONCLUSIONS: A two-step educational approach for patients on long-term oral anticoagulation does not improve TSTR percentages in the short term. PMID- 17115270 TI - Diagnosis of heavy metal contamination in agro-ecology of Gujranwala, Pakistan using cattle egret (Bubulcus ibis) as bioindicator. AB - The present study investigated the status of heavy metals: Lead (Pb), Cadmium (Cd), Chromium (Cr), Cobolt (Co), Silver (Ag) and Nickle (Ni) residues in egg, regurgitate and sediment samples collected from two colonies of cattle egret (Bubulcus ibis) during the breeding seasons between April and August in 2004 and 2005. The mean concentration of heavy metals in eggs and regurgitates was found higher compared to the maximum residue limit (mrl) standards prescribed by Environmental Protection Agency (EPA). A further comparison was made with a similar study conducted in China. High concentrations of heavy metals have contributed to the altered breeding behaviour of the bird species studied. Based on these findings we suggested a regular monitoring of the spread of these pollutants as these have not yet reached to the sediments. PMID- 17115271 TI - Predicting toxicity in marine sediment in Taranto Gulf (Ionian Sea, Southern Italy) using Sediment Quality Guidelines and a battery bioassay. AB - The goal of this study was to assess coastal marine pollution in the Mar Piccolo and Taranto Gulf (Ionian Sea, Southern Italy) by combining chemical and toxicological data in order to compare and integrate both approaches. Pollutants levels, traditionally, have limited ability to predict adverse effects on living resources. Moreover, in order to provide information on the ecological impact of sediment contamination on aquatic biota Numerical Sediment Quality Guidelines (SQGs) and sediment toxicity bioassays were carefully recommended. In this study ERL (effect range low)/ERM (effect range medium value) and TEL (threshold effect level)/PEL (probable effect level) guidelines have been used. Bioassays were performed with two species of amphipods Gammarus aequicauda and Corophium insidiosum, one species of isopod Idotea baltica and bivalve Mytilus galloprovincialis larvae. The TEL/PEL analysis suggested that, especially for stations 1 and 2, sediments in Mar Piccolo should contain acutely toxic concentrations of metals. In particular Hg content, in station 1, was about 17 times PEL value. 96 h LC(50) and 48 h EC(50) values were estimated for cadmium, copper and mercury in these species using the static acute toxicity test. M. galloprovincialis larvae was more sensitive than other species to all the reference toxicants tested (EC(50) determined for cadmium copper and mercury were of 0.59, 0.11 and 0.01 mg/l respectively). Significant differences in sensitivity of species tested to all reference toxicants (ANOVA p < 0.001) were recorded. Bioassays with these species allowed to estimated sediment toxicity from the different studied sites. On the basis of results obtained a good agreement was reported between chemical data and response of the biological endpoints tested. PMID- 17115272 TI - The interactive role of anxiety sensitivity and pubertal status in predicting anxious responding to bodily sensations among adolescents. AB - The present study examined the interaction between pubertal status and anxiety sensitivity (AS) in predicting anxious and fearful responding to a three-minute voluntary hyperventilation challenge among 124 (57 females) adolescents between the ages of 12 and 17 years (Mage = 15.04; SD = 1.49). As predicted, after controlling for baseline anxiety, age, and gender, there was a significant interaction between pubertal status and AS in predicting anxious responding to bodily sensations to the hyperventilation challenge. Specifically, adolescents reporting more advanced pubertal status and higher levels of AS reported the greatest post-challenge self-reported anxiety focused on bodily sensations, whereas pubertal status had relatively less of an effect on low AS adolescents. A test of specificity also was conducted; as expected, the interaction between AS and pubertal status was unrelated to generalized negative affectivity, suggesting the predictor variables interact to confer specific risk for anxious responding to bodily sensations. Finally, exploratory analyses of psychophysiological reactivity to the challenge indicated AS, but not pubertal status, moderated the relation between challenge-related change in heart-rate and post-challenge anxiety such that high AS youth who had experienced a relatively greater heart rate change reported the most anxious reactivity to the challenge. Results are discussed in relation to theory regarding vulnerability to anxious responding to bodily sensations among adolescents. PMID- 17115273 TI - Incorporating injured employee outcomes into physical and occupational therapists' practice: a controlled trial of the Worker-Based Outcomes Assessment System. AB - BACKGROUND: Work related musculoskeletal disorders (WRMSDs) remain costly. The Worker-Based Outcomes Assessment System (WBOAS) is an injury treatment improvement tool. Its purpose is to increase treatment effectiveness and decrease the cost of care delivered in Occupational Health Service clinics. METHODS: The study used a non-randomized (parallel cohort) control trial design to test the effects on injured employee outcomes of augmenting the standard care delivered by physical and occupational therapists (PT/OTs) with the WBOAS. The WBOAS works by putting patient-reported functional health status, pain symptom, and work role performance outcomes data into the hands of PT/OTs and their patients. Test clinic therapists were trained to incorporate WBOAS trends data into standard practice. Control clinic therapists delivered standard care alone. RESULTS: WBOAS augmented PT/OT care did improve (p< or =.05) physical functioning and new injury/re-injury avoidance and, on these same dimensions, cost-adjusted outcome. It did not improve (p>.05) mental health or pain symptoms or return-to-work or stay-at-work success nor, on these same dimensions, cost-adjusted outcome. CONCLUSION: Training PT/OTs to incorporate patient-reported health status, pain symptom, and work role performance outcomes trends data into standard practice does appear to improve treatment effectiveness and cost on some (e.g. physical functioning) but not other (e.g. mental health, pain symptoms) outcomes. PMID- 17115274 TI - An unusual pro-inflammatory role of interleukin-10 induced by arabinosylated lipoarabinomannan in murine peritoneal macrophages. AB - Various species of Mycobacteria produce a major cell wall-associated lipoglycan, called Lipoarabinomannan (LAM), which is involved in the virulence of Mycobacterial species. In this study, we tried to establish the role of the increased IL-10 secretion under Arabinosylated-LAM (Ara-LAM) treatment, the LAM that induces apoptosis in host macrophages or PBMC. We have studied the survival and apoptotic factors by western blotting, and estimated nitrite generation by Griess reaction, quantified iNOS mRNA by semi-quantitative RT-PCR, and ultimately the fate of the cells were studied by Flow Cytometric Analysis of AnnexinV-FITC binding. As per our observations, neutralization of released IL-10 in C57BL/6 peritoneal macrophages prior to Ara-LAM treatment, as well as macrophages from IL 10 knockout (KO) mice treated with Ara-LAM, showed significant down regulation of pro-apoptotic factors and up regulation of survival factors. These effects were strikingly similar to those when peritoneal macrophages were subjected to TNF alpha and IL-12 neutralization followed by Ara-LAM-treatment. However, under similar conditions virulent Mannosylated-LAM (from Mycobacterium tuberculosis) treatment of macrophages clearly depicts the importance of IL-10 in the maintenance of pathogenesis, proving its usual immunosuppressive role. Thus, from our detailed investigations we point out an unusual pro-inflammatory action of IL 10 in Ara-LAM treated macrophages, where it behaves in a similar manner as the known Th1 cytokines TNF- alpha and IL-12. PMID- 17115275 TI - Regioselectively modified sulfated cellulose as prospective drug for treatment of malaria tropica. AB - Adhesion of Plasmodium falciparum infected erythrocytes (IE) to placental chondroitin-4-sulfate (CSA) has been linked to the severe disease outcome of pregnancy-associated malaria. Consequently, sulfated polysaccharides with inhibitory capacity may be considered for therapeutic strategies as anti-adhesive drugs. During in vitro screening a regioselectively modified cellulose sulfate (CS10) was selected as prime candidate for further investigations because it was able to inhibit adhesion to CSA expressed on CHO cells and placental tissue, to de-adhere already bound infected erythrocytes, and to bind to infected erythrocytes. Similar to the undersulfated placental CSA preferred by placental binding infected erythrocytes, CS10 is characterized by a clustered sulfate pattern along the polymer chain. In further evaluation of its effects on P. falciparum interactions with host erythrocytes, we now show that CS10 inhibits the in vitro asexual growth of parasites in erythrocytes. Furthermore, we show that CS10 interferes with C1 of the classical complement pathway but not with MBL of the lectin pathway. In order to gain insights into the possible interactions of CS10 with known parasite receptors at the molecular level, we designed 3D structures of characteristic stretches of CS10. CS10 fragments with clustered sulfate groups showed complex patterns of hydrophobic and hydrophilic patches most likely suitable for interactions with protein binding partners. The significance of CS10 interactions with the complement system as well as its anti malarial effect for prospective drug application are discussed. PMID- 17115276 TI - Monocyte macrophage differentiation in vitro: Fibronectin-dependent upregulation of certain macrophage-specific activities. AB - Transendothelial migration of monocytes followed by their differentiation into macrophages involves interaction of monocytes with subendothelial matrix. The influence of extracellular matrix on monocyte-macrophage differentiation was studied using an in vitro model system with human PBMC maintained on different matrix protein substrata. Upregulation of macrophage specific marker activities such as endocytosis of modified proteins, changes in expression of cell surface antigen, and production of matrix metalloproteinases was studied. Cells maintained on Fibronectin (Fn) showed significantly higher rate of endocytosis and production of MMP2 and MMP9 when compared to other matrix protein substrata. Immunoblot analysis, ELISA, and zymography showed that Fn-dependent upregulation of MMPs was blocked by antibodies to alpha(5)beta(1) integrin indicating that the Fn effect was mediated by integrins. The Fn effect on mo-mPhi was blocked by genistein and herbimycin. As monocytes differentiate to macrophages there was an increase in the rate of production of Fn. These results indicate the influence of the microenvironment of the cell, particularly Fn, on mo-mPhi differentiation and integrin-mediated downstream signaling through focal adhesion kinase and Src type tyrosine kinase is involved in this. PMID- 17115277 TI - Immunological evaluation of urinary trypsin inhibitors in blood and urine: role of N- & O-linked glycoproteins. AB - Urinary trypsin inhibitors (uTi) suppress serine proteases during inflammation. After liberation from proinhibitors (P-alpha-I and I-alpha-I) by the white blood cell (WBC) response, uTi readily pass through the kidneys into urine. A key uTi, bikunin, is attached to O-linked and N-linked glycoconjugates. Recently, uTi inhibitors, called uristatins, were found to lack the O-linked glycoconjugates. Monoclonal antibodies were produced using purified uristatin and screened for binding differences to uristatin, bikunin, P-alpha-I, and I-alpha-I. Antibody binding patterns were characterized using immunoaffinity binding onto protein chip surfaces and analysis by Surface Enhanced Laser Desorption/Ionization mass spectrometry (SELDI), using specimens from patients and from purified uTi standards. Antibodies were developed and used in an enzyme-linked immunosorbent assay (ELISA) method for uTi measurement in urine and plasma specimens. ELISA was performed on specimens from normal, presumed healthy, controls and from patients who had been screened for inflammation using a high sensitivity C-reactive protein (CRP) test and a complete blood count (CBC). Polyclonal antibody against uTi showed cross-reactivity with the Tamm-Horsfall protein (THP) and with proinhibitors. Screening of anti-uTi monoclonal antibodies (Mab) revealed antibodies that did not cross-react with either of the above, thus providing a tool to measure both uristatin and bikunin in urine with Mab 3G5 and in plasma with Mab 5D11. The monoclonal antibody 5D11 cross-reacts with specific N-linked glycoconjugates of uristatin present in plasma. In ca 96% of healthy adults, uTi were present at <12 mg/l in urine and <4 mg/l in plasma. We also found that patients with an inflammation and a CRP of >2.0 mg/l had higher urinary concentrations of uTi than the control population in every subject. Free uristatin and bikunin pass readily into urine and are primarily bound to heavy chains that constitute the proinhibitor form in plasma. PMID- 17115279 TI - Profiling terminal N-acetyllactosamines of glycans on mammalian cells by an immuno-enzymatic assay. AB - Profiling of carbohydrate structures on cell membranes has been difficult to perform because of the complexity and the variations of such structures on cell surface glycans. This study presents a novel method for rapid profiling of cell surface glycans for terminal N-acetyllactosamines (Galbeta1-(3)4GlcNAc-R) that are uncapped, capped with sialic acid as SA-Galbeta1-(3)4GlcNAc-R, or with alpha1,3galactosyls as the alpha-gal epitope- Galalpha1-3Galbeta1-(3)4GlcNAc-R. This method includes two enzymatic reactions: (1) Terminal sialic acid is removed by neuraminidase, and (2) alpha-gal epitopes are synthesized on the exposed N acetyllactosamines by alpha1,3galactosyltransferase. Existing and de novo synthesized alpha-gal epitopes on cells are quantified by a modification of radioimmunoassay designated as "ELISA inhibition assay," which measures binding of the monoclonal anti-Gal antibody M86 to alpha-gal epitopes. This binding is proportional to the number of cell surface alpha-gal epitopes. The amount of free M86 antibody molecules remaining in the solution is determined by ELISA using synthetic alpha-gal epitopes linked to albumin as solid phase antigen. The number of alpha-gal epitopes on cells is estimated by comparing binding curves of M86 incubated with the assayed cells, at various concentrations of the cells, with the binding of M86 to rabbit red cells expressing 2 x 10(6) alpha-gal epitopes/cell. We could demonstrate large variations in the number of sialic acid capped N-acetyllactosamines, alpha-gal epitopes and uncapped N-acetyllactosamines on different mammalian red blood cells, and on nucleated cells originating from a given tissue in various species. This method may be useful for rapid identification of changes in glycosylation patterns in cells subjected to various treatments, or in various states of differentiation. PMID- 17115280 TI - Interaction of N-linked glycans, having multivalent GlcNAc termini, with GM3 ganglioside. AB - GM3 ganglioside interacts specifically with complex-type N-linked glycans having multivalent GlcNAc termini, as shown for (1) and (2) below. (1) Oligosaccharides (OS) isolated from ConA-non-binding N-linked glycans of ovalbumin, whose structures were identified as penta-antennary complex-type with bisecting GlcNAc, having five or six GlcNAc termini (OS B1, B2), or bi-antennary complex-type having two GlcNAc termini (OS I). OS I is a structure not previously described. (2) Multi-antennary complex-type N-linked OS isolated from fetuin, treated by sialidase followed by beta-galactosidase, having three or four GlcNAc termini exposed. These OS, conjugated to phosphatidylethanolamine (PE), showed clear interaction with (3)H-labeled liposomes containing GM3, when various doses of OS PE conjugate were adhered by drying to multi-well polystyrene plates. Interaction was clearly observed only with liposomes containing GM3, but not LacCer, Gb4, or GalNAcalpha1-3Gb4 (Forssman antigen). GM3 interaction with PE conjugate of OS B1 or B2 was stronger than that with PE conjugate of OS I. GM3 interacted clearly with PE conjugate of N-linked OS from desialylated and degalactosylated fetuin, but not native fetuin. No binding was observed to cellobiose-PE conjugate, or to OS-PE conjugate lacking GlcNAc terminus. Thus, GM3, but not other GSL liposomes, interacts with various N-linked OS having multiple GlcNAc termini, in general. These findings suggest that the concept of carbohydrate-to-carbohydrate interaction can be extended to interaction of specific types of N-linked glycans with specific GSLs. Natural occurrence of such interaction to define cell biological phenomena is under investigation. PMID- 17115281 TI - Differential surface expression and possible function of 9-O- and 7-O-acetylated GD3 (CD60 b and c) during activation and apoptosis of human tonsillar B and T lymphocytes. AB - The disialoganglioside GD3 (CD60 a) and its O-acetylated variants have previously been described as surface molecules of human T lymphocytes of the peripheral blood system. Here we report the expression of the 9-O-, and 7-O-acetylated disialoglycans of GD3 (CD60 b and CD60 c respectively) on human tonsillar lymphocytes. CD60 b and c are surface-expressed on activated germinal centre B cells and colocalize in raft-like structures on the cell surface together with the cytoplasmic tyrosine kinase Lyn and Syk. Addition of CD60 b and c mAb together with anti-IgM/IL-4 to in vitro cultivated tonsillar B cells resulted in a costimulatory effect. During spontaneous and staurosporine-induced apoptosis a distinct population of activated annexin V+/CD60 b+/CD60 c- B cells was observed. CD60 b and c are also found on cells of the extrafollicular T cell area. On tonsillar T cells, CD60 b mAb had a costimulatory effect together with PHA while CD60 c mAb alone was sufficient to induce proliferation. In further contrast to B cells, during apoptosis a distinct CD60 b+ T cell subpopulation was not observed. Together, surface-expressed CD60 b and c are differently expressed on tonsillar B and T cells and may be involved in the regulation of activation and apoptosis of lymphocytes in secondary lymphatic tissue. PMID- 17115282 TI - An analyst's experience working in a skilled nursing facility: a case study. AB - With the closing of long-term psychiatric facilities, no alternative social institution has arisen to treat those persons whose chronic mental illness makes self-care impossible. This paper explores the typical paths to placement in a skilled nursing facility or nursing home and the author's experience in working with this population using the principles and techniques of interpersonal psychoanalysis. She presents several case examples and explicates both the analyst's and the patient's transference. The author discusses the difficulties inherent in creating an analytic space in such a setting and working outside of the usual psychoanalytic frame. PMID- 17115283 TI - From neurotic guilt to existential guilt as grief: the road to interiority, agency, and compassion through mourning. Part II. AB - This study is a continuation of an article that was published in the previous issue of The American Journal of Psychoanalysis (Volume 66, Number 3, September 2006). The case of Helen is illustrated in terms of the analysand's developmental mourning, countertransference, interiority, antilibidinal ego, and transformation of aggression into self-agency. PMID- 17115284 TI - A competency-based approach to managing violence with involuntary outpatient treatment. AB - Involuntary outpatient treatment is one of the most controversial areas in public psychiatry. There are cogent arguments and strong emotions both for and against the use of it. Yet there is violent behavior towards others by individuals with mental illness who reside in the community that is not managed well even when recognized as highly likely. For individuals already in the community mental health system, the ability to keep them in treatment, even against their will, is necessary in some instances to decrease the likelihood of them engaging in outwardly directed violent behavior. PMID- 17115285 TI - Expression of VEGFR3 in glioma endothelium correlates with tumor grade. AB - Angiogenic processes are regulated by vascular endothelial growth factors (VEGFs) and their receptors VEGFR1 (Flt-1), 2 (Flk-1) and 3 (Flt-4). While VEGFR2 is thought to play a central role in tumor angiogenesis, anti-angiogenic therapies targeting VEGFR2 in glioma models can show escape phenomena with secondary onset of angiogenesis. The purpose of this study was to find explanations for these processes by searching for alternative pathways regulating glioma angiogenesis and reveal a correlation with tumor grade. Thus, VEGFR3, which is not expressed in normal brain, and its ligands VEGF-C and -D, were assessed in high grade (WHO degrees IV, glioblastomas, GBM) and low grade gliomas [WHO degrees II astrocytomas (AII)]. In all GBM, a strong protein expression of VEGFR3 was found on tumor endothelium, VEGF-C and -D expression was found on numerous cells in areas of high vascularization. On RNA level, a significant up-regulation of VEGFR3 was detected in GBM compared to AII and non-neoplastic brain. In AII, only very moderate VEGFR3, VEGF-C and -D expression was found on protein and RNA level indicating a correlation of VEGFR3 expression with tumor grade. VEGFR3 signal in both grades was found predominantly on endothelial cells, confirmed by VEGFR3 expression on isolated CD31 positive cells and the expression of various endothelial markers on VEGFR3-positive cells isolated from GBM. The demonstration of a complete angiogenic signaling system that is dependent on tumor grade may influence the traditional paradigm of glioma angiogenesis and may provide a basis for more effective anti-angiogenic treatment strategies. PMID- 17115287 TI - Caregivers' perceptions of child mental health needs and service utilization: an urban 8-year old sample. AB - This study extended previous research on mental health utilization to a sample of 214 preadolescent children and their caregivers. Predictors of two distinct phases of service utilization were tentatively examined in multivariate analyses: caregivers' perceptions that children needed services and receipt of those services by children. Of these children, 24.8% were perceived by their caregivers as needing mental health services; 11.7% received mental health services and 13.1% did not. Internalizing behavioral problems increased children's likelihood of being seen as needing services, but failed to increase likelihood of receiving services. Four factors predicted receipt of mental health services: relative lack of poverty, non-African American ethnicity, externalizing behavior problems, and child history of maltreatment. Implications for services targeted at preadolescent children are discussed. PMID- 17115286 TI - Child sociodemographic characteristics and common psychiatric diagnoses in medicaid encounter data: are they valid? AB - This study describes the rate that Medicaid encounter data on gender, race/ethnicity, and diagnosis matched information in the medical record, among a statewide sample of Medicaid children who received ongoing care for attention deficit hyperactivity disorder (ADHD), conduct disorder (CD), and major depression (MD) in outpatient specialty mental health clinics in 1998-1999. The match rate for gender was 99%; and for race/ethnicity it was 71.8%, 90.5%, and 89.7% for Caucasian, African American, and Hispanic children, respectively. Misidentified Caucasian children were more likely to be recorded as African American or Hispanic than misidentified minority children to be recorded as Caucasian. Diagnosis match rates were high (ADHD: 98%, CD: 89%, MD: 89%). If the California Department of Mental Health relied solely on Medicaid encounter data, misclassification of African American or Hispanic children as Caucasian could produce an underestimate of their service use. PMID- 17115288 TI - Marginal zone lymphoma of both spleen and kidney displaying transformation into large B-cell lymphoma. AB - We report a case of simultaneous involvement of the spleen and the left kidney in a marginal zone lymphoma with a monotypic lymphoplasmacytic cell component, which transformed into a diffuse large B-cell lymphoma of the immunoblastic type. PCR showed that the small and large B-cell populations carried the same type of immunoglobulin heavy chain gene rearrangement. This type of rearrangement was detected in the spleen, the latero-aortic lymphadenopathy and the kidney demonstrating that it is the same lymphoma that affected both organs and the lymph nodes. Primary renal lymphoma is very rare and only a few cases of renal marginal zone lymphoma, MALT type, have been reported. Involvement of simultaneous multiple sites has been described in MALT type lymphoma, but splenic involvement secondary to renal MALT lymphoma seems to have never been observed. Nevertheless, in our case the huge size of the spleen associated with splenic hilar node involvement is consistent with primary splenic marginal zone lymphoma. The extension into latero-aortic lymph nodes of this lymphoma can explain secondary kidney involvement. The nodal Kaposi's sarcoma observed in this patient of Mediterranean origin was probably coincidental. PMID- 17115289 TI - Eosinophilic cystitis simulating invasive bladder cancer: a real diagnostic challenge. AB - OBJECTIVES: To report our experience with eosinophilic cystitis (EC) presented as invasive bladder cancer. MATERIALS AND METHODS: We recently treated three patients with bladder masses mimicking neoplasms and biopsy proved the diagnosis of EC. Data from our three patients were presented. RESULTS: There were two females and one male with ages of 14, 43 and 38 years. All the three patients had irritative bladder symptoms, suprapubic pain and hematuria. Bladder mass was detected by pelvic examination in the three patients and confirmed by radiologic tests and cystoscopy. In one patient, the mass caused bilateral hydroureteronephrosis while the upper tract was normal in the other two. Elevated serum leukocytes was evident in the three cases while peripheral eosinophilia was observed in one. Biopsy showed EC in all the three patients who were treated by transurethral resection of the lesions followed by a combination of corticosteroids, antibiotics and antihistaminics. All patients experienced marked improvement during a follow-up duration up to 30 months. CONCLUSION: EC is a rare disease. In addition to symptoms of frequency, dysuria, hematuria and suprapubic pain, the disease may present with a bladder mass mimicking invasive bladder cancer. Resection of the lesion is mandatory with systemic treatment of corticosteroids, antihistaminics and antibiotics. Early detection and prompt treatment usually result in a good prognosis. PMID- 17115290 TI - Surgical management of bilateral ureteral endometriosis. AB - Ureteral endometriosis is a rare disease that typically is unilateral. Endometriosis involving both ureters and surgical management after hormone therapy failure has seldom been described. We describe a patient with bilateral ureteral endometriosis who underwent ureteroneocystostomy with psoas hitches of both ureters. A 33-year-old woman with advanced endometriosis and recurrent pyelonephritis was found to have high-grade bilateral ureteral obstruction at the pelvic inlet from ureteral endometriosis. The patient subsequently underwent a supracervical hysterectomy with bilateral salpingo-oophorectomy, ureterolysis, and ureteroneocystostomy with psoas hitches and ureteral stent placements. Surgical therapy is reserved for advanced disease with the optimal choice being a ureteral reimplantation with a psoas hitch. The key operative point for a successful psoas hitch ureteral reimplantation is completely mobilizing the bladder anteriorly and laterally. PMID- 17115291 TI - Common symptoms in men with prostatic inflammation. AB - PURPOSE: We evaluated 96 patients with prostatic inflammation in terms of their symptoms and aimed to find common types and frequencies of symptoms in these patients. PATIENTS AND METHODS: The mean age of the patients was 38.0+/-8.7 (range 21-58) years. Physical examination, digital rectal examination, microscopic prostatic secretion assessment and urine cultures after taking a detailed medical history were performed. Urine samples before and after prostatic massage were collected for urine culture. Frequency and types of patients' symptoms were evaluated. All patients were asked about lower urinary tract symptoms, sexual dysfunction and other complaints. RESULTS: Lower urinary tract symptoms and lumbal pain were more prevalent in elder patients. Ejaculation disorder was the most common sexual problem (n=65, 67.7%). Erectile dysfunction and decreased libido were observed in 29 (30.2%) and 22 (22.9%) of the patients. Other complaints were lumbal pain (n=34, 35.4%), perineal fullness (n=50, 52.1%), haemospermia (n=20, 20.8%) and scrotal pain (n=43, 44.8%). CONCLUSION: Prostatic inflammation was usually seen in men of the third and fourth decade. Sexual dysfunction was the most common symptom in this particular group of patients. PMID- 17115292 TI - Bladder leiomyosarcoma in a boy. AB - Leiomyosarcoma of the urinary bladder in childhood is uncommon. We reported bladder leiomyosarcoma in a 10-year-old boy. He had underwent the extirpation of the tumor and performed adjuvant chemotherapy in combination with radiotherapy according to the Intergroup Rhabdomyosarcoma Study-4 regimens. He had no evidence of recurrence of disease after 3 years. PMID- 17115293 TI - Brucella orchitis: a rare cause of testicular mass: report of a case. AB - Brucellosis is a multiorgan infectious disease. The genitourinary system is affected in 2-20% of the cases and the most common form is orchitis. Rarely, patients may present with a testicular mass and it must be distinguished from malignant processes. We report brucellar orchitis, a rare cause of testicular mass, in a 22-year-old man. We described the clinicopathological features of this rare entity and reviewed the literature. PMID- 17115294 TI - Familial torsion of the testis. AB - We report a family of a man and two of his three male children who all had torsion of the left testis, all at the age of 15 years and all occurring during sleep. Their management and the possible modes of inheritance are discussed. PMID- 17115295 TI - The effect of two different doses comprising the simultaneous administration of intravenous B-complex vitamins and oral folic acid on serum homocysteine levels in hemodialysis patients. AB - BACKGROUND: Several regimens using different doses of folic acid (FA) alone or supplemented with B-complex vitamins (BCVs) have been tested for their ability to reduce total homocysteine (tHcy) serum levels in hemodialysis (HD) patients. In the present study, we assessed the effect of two different doses comprising the simultaneous administration of intravenous (IV) BCVs and an oral FA supplementation on serum tHCy levels in HD patients. PATIENTS-METHODS: In a cohort of 49 patients (31 male, 18 female) undergoing chronic HD treatment for a mean of 40.0+/-40.7 months, serum concentrations of tHcy, folate and vitamin-B12 (vB12) were determined at the end of three sequential periods as follows: 20 weeks without any BCV and/or FA supplementation (period A), 20 weeks with a dose comprising the simultaneous administration of IV BCVs and an oral supplementation of 5 mg of FA once a week (period B), and 20 weeks with a dose comprising the simultaneous administration of IV BCVs and an oral supplementation of 5 mg of FA thrice a week (period C). An IV dose of BCVs consisting of a 5 mL solution containing vitamin B1 (250 mg), vitamin B6 (250 mg) and vitamin B12 (1.5 mg) was administered at the end of hemodialysis. RESULTS: Mean serum tHcy levels were significantly higher at the end of period A relative to levels at the end of periods B and C (35.8+/-23 micromol/L vs. 22.0+/-17.6 and 15.0+/-4.5 micromol/L, respectively; p<0.000001). Mean serum folate levels and mean serum vB12 levels were significantly lower at the end of period A relative to levels at the end of periods B and C (p<0.000001). Mean serum tHcy levels were lowest at the end of period C (p<0.000001 in comparison to periods A and B), and 26 of the 49 HD patients (67.3%) possessed tHcy levels below 16 micromol/L. CONCLUSIONS: In HD patients, high doses consisting of the simultaneous administration of IV BCVs and an oral FA supplementation resulted in the efficient reduction of serum tHcy levels. PMID- 17115296 TI - Case series: adult testicular dermoid tumours--mature teratoma or pre-pubertal teratoma? AB - Adult testicular dermoid tumours are rare tumours with no reported potential for recurrent or metastatic spread. Despite this they are currently classified as mature teratoma and managed as if they have equivalent malignant potential. This report describes two cases of adult mature teratoma of dermoid type and questions the classification and pathogenesis of this disease. In one of the cases there was a clear history of a testicular lump arising pre-pubertally, raising the possibility that some adult dermoid tumours may in fact be pre-pubertal teratomas that have persisted into adulthood. Classification as a mature teratoma carries with it a follow-up regimen that includes numerous radiological investigations with their attendant radiation exposure. A positive histological diagnosis and separate classification of adult dermoid tumours would allay clinical fears of recurrence and metastasis and negate the need for repeated radiological investigations. PMID- 17115297 TI - The management of hydronephrosis in patients undergoing TURBT. AB - OBJECTIVE: Previous studies have shown the negative prognostic correlation of hydronephrosis in bladder cancer; however, practical uncertainties remain regarding the management of these patients. METHODS: We retrospectively reviewed the notes of patients undergoing TURBT over a three year period and recorded the management and outcome of patients with hydronephrosis. RESULTS: Six percent with bladder cancer had hydronephrosis. Nearly all the cases had muscle invasive disease. At TURBT, the ureteric orifice was seen in 41%; in the remaining 59% of patients, the ureteric orifice was involved and resected. This resolved the hydronephrosis in only one patient (who had superficial disease). CONCLUSIONS: Hydronephrosis in bladder cancer is associated with a poor prognosis. The hydronephrosis does not resolve with resection alone. As awaiting it's resolution may delay definitive treatment, we suggest aggressive management of hydronephrosis from the time of initial diagnosis with ureteric stenting in order to protect renal units and optimize renal function prior to further definitive treatment of bladder cancer. PMID- 17115298 TI - Giant condylomata accuminata of scrotum representing as carcinoma (a case report). AB - In this case report a 51-year-old man admitted with a large warty lesion of scrotum representing as carcinoma was presented. A wide surgical resection of the lesion was done with the hystopathologic diagnosis of condyloma accuminata. He was followed for 12 months with satisfactory functional and cosmetic result with no recurrence with the time being. PMID- 17115299 TI - Management of renal abscess formation after embolization due to renal angiomyolipomas in two cases. AB - Current management strategies of renal angiomyolipomas (AMLs) include observation, embolization and partial or total nephrectomy. Selective arterial embolization is an effective and safe treatment of large angiomyolipomas with low complication rates. Percutaneous drainage was the recommended treatment for renal abscess formation following the embolization. Herein we describe two cases which we performed percutaneous drainage of the liquefaction of entire tumors after embolization. Open surgery was needed for one of the patients who showed recurrence after percutaneous drainage and alcohol irrigation of the cavity, whereas percutaneous drainage was the sufficient treatment for the other patient as recommended. PMID- 17115301 TI - Sobemoviruses possess a common CfMV-like genomic organization. AB - Based on structural differences in the ORF2 region, the sobemoviruses have been subdivided into southern cowpea mosaic virus (SCPMV)-like and cocksfoot mottle virus (CfMV)-like types of genome organization. However, nearly identical amino acid sequences are encoded by these subgroups in different reading frames of ORF2, suggesting that insertion or deletion of appropriate nucleotides could restore similar genomic organizations for these viruses. We resequenced the regions of inconsistency for isolates of four SCPMV-like viruses: lucerne transient streak virus, ryegrass mottle virus, southern bean mosaic virus, and SCPMV. A comparison of nucleic acid composition of these sequences with previously published ones revealed crucial differences that established a common CfMV-like genomic organization for these sobemoviruses. PMID- 17115300 TI - Transcriptomic adaptations in rice suspension cells under sucrose starvation. AB - Sugar is an important resource for energy generation and developmental regulation in plants, and sucrose starvation causes enormous changes in cellular morphology, enzyme activities and gene expression. Genome-wide gene expression profiling provides a comprehensive knowledge of gene expression under nutrient depletion and senescence; however, that of a monocot model plant, rice, under sucrose depletion is still under investigation. Here, the time-course monitoring of gene expression profiles in sucrose-starved rice (Oryza sativa cv Tainung67) suspension cells was investigated by 21495 probes contained in Agilent rice chip. In sucrose-starved cells, the induced vacuolar biogenesis coincided with significantly upregulated transcripts of H+-pyrophosphatase, delta-TIP, one putative alpha-TIP, several vacuolar proteases and proteinase inhibitors, and one OsATG3. To survey the overall metabolic adaptations under sucrose depletion, the genes with significantly altered expression level were incorporated into multiple metabolic pathways. Most genes encoding enzymes involved in biosynthesis and degradation pathways of various macromolecules were comprehensively down-and upregulated, respectively, with sucrose starvation. Transcriptional regulation of gene expression is important for physiological adaptations to environmental stress, and many transcription factors, including bZIPs, NACs, and WRKY, showed significant increase in transcriptional level under sucrose starvation. Concurrently, statistical analysis revealed that their corresponding consensus cis-elements, such as ABA-responsive element, CACG, ACI, ACII and CTTATCC, were frequently found in the promoter regions of many sucrose starvation-upregulated genes. Particle bombardment-mediated and luciferase activity-based transient promoter assays revealed the CTTATCC, derived form TATCCA, and the AC motifs to be promising sucrose-starvation responsive activators in rice suspension cells. PMID- 17115302 TI - Simian varicella virus gene 61 encodes a viral transactivator but is non essential for in vitro replication. AB - Simian varicella virus (SVV) is closely related to varicella-zoster virus (VZV), the causative agent of chickenpox and shingles. The SVV and VZV gene 61 polypeptides are homologs of the HSV-1 ICP0, a viral transactivator which appears to play a role in viral latency and reactivation. In this study, the molecular properties of the SVV 61 were characterized. The SVV open reading frame (ORF) 61 encodes a 54.1-kDa polypeptide with 37% amino acid identity to the VZV 61. Homology to the HSV-1 ICP-0 is limited to a conserved RING finger motif at the amino terminus of the protein. A nuclear localization sequence (nls) at the carboxy-terminus directs the SVV 61 to the cell nucleus, while a SVV 61nls(-) mutant is confined to the cell cytoplasm. The SVV 61 transactivates its own promoter as well as SVV immediate early (IE, ORF 62), early (ORFs 28 and 29), and late (ORF 68) gene promoters in transfected Vero cells. The RING finger and nls motifs are required for efficient SVV 61 transactivation. The SVV 61 has no effect on the ability of the major SVV transactivator (IE62) to induce SVV promoters. Generation and propagation of a SVV gene 61 deletion mutant demonstrated that the SVV 61 is non-essential for in vitro replication. SVV gene 61 is expressed in liver, lung, and neural ganglia of infected monkeys during acute simian varicella. PMID- 17115303 TI - Sequencing of the bicistronic genome segments S7 and S9 of Mal de Rio Cuarto virus (Fijivirus, Reoviridae) completes the genome of this virus. AB - The nucleotide sequences of genomic segments S7 and S9 of Mal de Rio Cuarto virus (MRCV, Fijivirus group II) have been determined, thus completing the entire genome sequence of the virus. These segments showed a non-overlapping bicistronic structure, as in other members of the genus. MRCV S7 ORF-1 had a length of 1086 bp and encoded a 41.5 kDa putative polypeptide, whereas MRCV S7 ORF-2 had a length of 930 bp and encoded a 36.8 kDa putative polypeptide. Proteins of 39 and 20.5 kDa were predicted for the 1014 bp long MRCV S9 ORF-1 and the 537 bp long MRCV S9 ORF-2, respectively. The terminal 5' and 3' sequences of both segments were 5'AAGUUUUU3' and 5'CAGCUnnnGUC3', respectively. Specific imperfect inverted repeats of each segment were identified. Comparison of the predicted proteins with those of related virus genome segments counterparts in maize rough dwarf virus (MRDV) and rice black streaked dwarf virus (RBSDV), showed 64.5-44.3% identities. These values are lower than those resulting from comparisons between MRDV and RBSDV. The topology of the trees obtained using the complete nucleotide and amino acid sequences of MRCV S7 and MRCV S9 was consistent with the analysis of the other MRCV segments previously published. PMID- 17115304 TI - Analysis of hepatitis B surface antigen mutations in Mongolia: molecular epidemiology and implications for mass vaccination. AB - Although the potential significance of hepatitis B surface antigen (HBsAg) mutants for failure of immunization has been studied in some endemic countries, whether the "a" determinant variants are responsible for vaccine failure in Mongolia remains unknown. Fifty-nine HBsAg-positive children (age: 8.8 +/- 0.9 years) who had been observed during the nationwide survey of vaccinated cohorts conducted in 2004 were subjected to molecular analyses of hepatitis B virus (HBV). Partial S gene sequences encoding amino acids (aa) 40-171 of HBsAg were determined in 57 children (96.6%) who had detectable HBV DNA. Phylogenetic analysis of the S gene sequences revealed that genotype D accounted for 93.0% and genotype A for 5.3%. Only one child (1.7%) had HBVs of genotypes A and D. HBsAg mutations were found in 17 (29.8%) children ranging from 1 to 4 aa per subject (mean +/- SD, 1.6 +/- 0.9 aa). Pro127Thr and Thr118Ala were the most common substitutions, which occurred in 6 (10.5%) and 3 (5.3%) subjects, respectively; none had Gly145Arg. There were no significant associations in the prevalence of HBsAg mutations with age, sex, residential area, or vaccination status against hepatitis B. Analysis of the deduced amino acid sequence of the entire preS1/preS2/S gene revealed that eight genotype D isolates and one genotype A isolate were quite similar to previously-reported wild-type isolates, suggesting that they are essentially wild-type, but not vaccine-induced mutants. In conclusion, the results demonstrate that hepatitis B surface gene mutants do not play a significant role in vaccination failure in Mongolia. PMID- 17115305 TI - Genome characterisation of the newly discovered avian influenza A H5N7 virus subtype combination. AB - In Denmark, in 2003, a previously unknown subtype combination of avian influenza A virus, H5N7 (A/Mallard/Denmark/64650/03), was isolated from a flock of 12,000 mallards. The H5N7 subtype combination might be a reassortant between recent European avian influenza A H5, H7, and a third subtype, possibly an H6. The haemagglutinin and the acidic polymerase genes of the virus were closely related to a low-pathogenic Danish H5N2 virus A/Duck/Denmark/65041/04 (H5N2). The neuraminidase gene and the non-structural gene were most similar to the highly pathogenic A/Chicken/Netherlands/1/03 (H7N7) and the human-fatal A/Netherlands/219/03 (H7N7), respectively. The basic polymerase 1 and 2 genes were phylogenetically equidistant to both A/Duck/Denmark/65047/04 (H5N2) and A/Chicken/Netherlands/1/03 (H7N7). The nucleoprotein and matrix gene had highest nucleotide sequence similarity to the H6 subtypes A/Duck/Hong Kong/3096/99 (H6N2) and A/WDk/ST/1737/2000 (H6N8), respectively. All genes of the H5N7 strain were of avian origin, and no further evidence of pathogenicity to humans has been found. PMID- 17115306 TI - Rectovaginal fistula after STARR procedure complicated by haematoma of the posterior vaginal wall: report of a case. AB - We report the case of a patient treated with the stapled transanal rectal resection (STARR) procedure for obstructed defecation, who developed an early postoperative haematoma of the posterior vaginal wall and, after 30 days, a rectovaginal fistula (RVF), even though the intervention had been performed according to the standardized technique. After clinical examination and three dimensional anal endosonography, we carried out a successful surgical correction with double vaginal and rectal flaps with repair of the rectovaginal septum and without faecal diversion. The STARR procedure, even if performed according to a rigorous application of the methodological standards, may be followed by a RVF possibly due to a blood collection leading to ischaemia of the vaginal wall. PMID- 17115307 TI - Endoanal ultrasonography in the diagnosis and operative management of perianal endometriosis: report of two cases. AB - We report two cases of perianal endometriosis in which we were greatly assisted by endoanal ultrasonography. Patient 1 was a 43-year-old woman with perianal pain. Endosonography showed a hypoechoic mass in the anterior perianal region without involvement of the anal sphincter. Local excision was performed under spinal anesthesia without damage to the anal sphincter. Patient 2 was a 30-year old woman with perianal pain coinciding with her menstrual period. Endosonography showed a heterogeneous mass containing cystic anechoic areas in the right anterior perianal region and involving the external anal sphincter. Wide excision, including the episiotomy scar and part of the external anal sphincter, and primary sphincteroplasty were performed under spinal anesthesia. According to our experience, preoperative endosonography is a reliable technique for visualizing perianal endometriosis and for diagnosing anal sphincter involvement. Operative management should be determined on the basis of preoperative and intraoperative ultrasonographic assessment. PMID- 17115308 TI - Colonic perforation as a complication of ventriculoperitoneal shunt: a case report. AB - Late perforation after ventriculoperitoneal (VP) shunting is extremely rare. Colonic perforation is uncommon and represents 0.1%-0.7% of abdominal complications. Colonic perforation can challenge diagnostic and therapeutic decisions, and there are no clear guidelines on the management of this problem. We present a 34-year-old woman who was admitted for a 1-week history of sensation of a foreign body through the anus at the time of bowel movements. She had previously undergone a VP derivation for hydrocephalus secondary to neurocysticercosis. Plain abdominal radiographs demonstrated the shunt within the colonic lumen and through the descendening and sigmoid colon. The shunt was exteriorized in the cervical area and a laparotomy was performed with a primary two-layer colonic close. The patient received antibiotic therapy for 2 weeks with good outcome. Percutaneous and endoscopic approaches have been reported in patients with no abdominal signs. Prompt recognition of this complication is critical to avoid high mortality rates. PMID- 17115309 TI - Laparoscopic combined rectal anterior resection and total hysterectomy with bilateral salpingo-oophorectomy. AB - Pelvic conditions involving both the colorectum and gynaecological organs are traditionally treated by laparotomy. We report two cases of colorectal cancer and one of endometriosis treated by laparoscopic anterior resection and total hysterectomy bilateral salpingooophorectomy (LapAR & THBSO), at the same session. There have been no previously reported cases of the feasibility of such combined procedures, safely performed. Our results confirm that LapAR & THBSO is feasible and offers the advantages of a laparoscopic procedure in the hands of a well trained laparoscopic colorectal surgeon and gynaecologist working together. PMID- 17115310 TI - Long-acting octreotide in the treatment of diarrhea after pelvic pouch surgery. AB - After pelvic pouch surgery (restorative proctocolectomy), periodic or continuous diarrhea is common. Distressing diarrhea may be triggered by pouchitis, cuffitis or an anastomotic stricture. Medical therapy with loperamide or diphenoxylate is often unsatisfactory even after the correction of the underlying problem. Seven patients, who earlier had undergone restorative proctocolectomy, were treated with a standard dose of 10 mg of long-acting octreotide (Sandostatin LAR) for prolonged and distressing diarrhea which had not responded to conventional medication. Five of the patients had complete relief of diarrhea. The effect lasted for at least two months after a single dose. Two patients did not respond. No sideeffects were reported. Long-acting octreotide is effective in the treatment of severe diarrhea in patients after pelvic pouch surgery. PMID- 17115311 TI - Gasless transanal endoscopic surgery for rectal adenomas and early carcinomas. AB - Transansal endoscopic microsurgery (TEM) allows the excision of rectal tumors but is difficult and complex. A new TEM modification simplifying the procedure and reducing its cost is required. Between 1999 and 2004, 128 patients (78 women and 50 men) underwent gasless transanal endoscopic surgery (GTES) without use of a completely closed system by means of laparoscopic equipment. Histological examination revealed 112 adenomas, including 15 with neoplastic changes (6 cases of Tis and 9 cases of T1G1G2), 12 adenocarcinomas (1 case of Tis, 8 cases of T1G1G2, and 1 case of T2G1G2) and 3 carcinoid tumors. There was no operative mortality. One patient had postoperative bleeding. Adenomas recurred in 9 patients (8.3%). The adenocarcinomas demonstrated neither local recurrences nor distal spread. GTES may be considered a safe and effective minimally invasive treatment for patients with large adenomas and early carcinomas of the rectum. PMID- 17115312 TI - Improved detection of colorectal neoplasms with selective use of chromoendoscopy in 2005 consecutive patients. AB - BACKGROUND: Colorectal cancer mortality is decreased by endoscopic polypectomy, but conventional colonoscopy may be inadequate for detecting subtle colonic lesions. METHODS: We selectively performed chromoendoscopy in all patients undergoing colonoscopy between January 1999 and December 2005 at the International Health Union of Rome. Patients with a history of colorectal polyps, inflammatory bowel disease, colorectal surgery or coagulopathy and those with poor bowel preparation were excluded from this analysis. Whenever colonoscopy revealed suspicious mucosal areas, dye-spraying with 0.2% indigo carmine solution was also performed. Findings from conventional and dyespraying views were classified morphologically, and specimens were analyzed histologically. Non adenomatous lesions were classified as negative findings. RESULTS: A total of 2005 patients underwent conventional colonoscopy and in 305 cases (15%) chromoendoscopy was also performed. Conventional colonoscopy identified 508 neoplasms in 381 patients (19%). Selective chromoendoscopy found an additional 244 neoplasms in 212 patients (11%). Thus, chromoendoscopy was positive in 212 (70%) of 305 patients in whom the examination was performed. Overall, 56 large, ulcerated, advanced cancers and 696 non-advanced neoplasms were found. Of the 696 nonadvanced neoplasms, 448 (65%) were polypoid and 248 (35%) were non-polypoid. All but 4 non-polypoid lesions were only detected with chromoendoscopy. Of the 248 non-polypoid lesions, 12 (5%) were depressed and 236 (95%) were flat. Advanced histology was present in 39 non-polypoid lesions (15%) and was more common in depressed lesions than in flat ones (58% vs. 13%; p<0.001). CONCLUSIONS: Our study confirms the existence of flat and depressed neoplasms in an Italian population. The vast majority of non-polypoid lesions were only detected by chromoendoscopy, and many lesions with advanced histology were missed by conventional colonoscopy. We therefore recommend selectively performing chromoendoscopy when conventional colonoscopy provides clues for non-polypoid lesions. Therefore, endoscopists should be trained in the detection of these subtle mucosal clues, as well as in the use of chromoendoscopy to enhance their detection. PMID- 17115313 TI - Bioabsorbable staple-line reinforcement to reduce staple-line bleeding in the transection of mesenteric vessels during laparoscopic colorectal resection: a pilot study. AB - BACKGROUND: Staple line hemorrhage and leak are the 2 most dreaded complications of laparoscopyassisted resection of colorectal cancer (LARCC). Recently, anastomotic staple lines have been reinforced with a range of absorbable and non absorbable bioprosthetic reinforcement materials. This pilot prospective study used the bioabsorbable Seamguard (BSG; WL Gore & Associates, Flagstaff, AZ) for routine reinforcement of the mesenteric vascular staple transection to prevent mesenteric hemorrhage and to assess its safety and feasibility in a consecutive series of laparoscopic colonic resections. METHODS: Twenty-five patients consecutively scheduled to undergo LARCC were enrolled in the study. All operations were performed with a standard LARCC technique which included loading of the BSG sleeves onto the jaws of the cutter/stapler. RESULTS: There were 23 wholly LARCC cases and 2 open conversions. BSG was used in all 25 patients. No patient experienced staple-line bleeding or other complications during the surgical procedure. The mean number of staple-line reinforcement sleeves used was 2.6 (range, 2-4). The mean operative time was 118 minutes (range, 65-184 minutes). Additional measures to achieve hemostasis were not required in any case. The mean cost was US 475.20 dollars for the procedure. CONCLUSIONS: This small pilot study has shown the routine use of BSG mesenteric staple line reinforcement to be safe, quick and effective during LARCC. PMID- 17115314 TI - Low hemorrhoidopexy staple line does not improve results and increases risk for incontinence. AB - BACKGROUND: The ability of stapled hemorrhoidopexy (SH) to cure hemorrhoidal symptoms appears to depend on patient characteristics and operative technique. We assessed the association between outcome of SH and patients' characteristics and procedure parameters (associated procedure, suture line height, doughnut size, presence of malpighian tissue or smooth muscle in specimen). METHODS: A total of 68 consecutive patients (56 males) were prospectively operated by 3 different surgeons. Hemorrhoids were grade II (6%), grade III (76%) or grade IV (18%). RESULTS: At a mean 32-week follow-up (range, 9-77), symptoms had resolved in 77% of patients, independently of any operative or clinical parameter. New onset anal incontinence occurred in 11 men (17%): all had urgency, with flatus and liquid stool incontinence in two, and flatus incontinence and mucus soiling in one. Univariate analysis revealed that persistent incontinence was associated with a staple line <6.5 mm from the dentate line, doughnut height <22 mm, and congestive external hemorrhoids; it was also operator dependent (p<0.05). At the 4-week follow-up, 19% of patients had persisting symptoms but only 8% had a demonstrable mucosal prolapse. CONCLUSION: Although the success rate of SH may not be influenced by technical variations, risk for moderate incontinence is elevated when the stapled line is low. PMID- 17115315 TI - The mesorectum: hypothesis on its evolution. AB - BACKGROUND: Two points are controversial in the anatomy of the mesorectum: (1) its origin; and (2) the existence of the lateral ligaments. We studied these structures in animals and in human fetuses. METHODS: Dissections were performed on quadrupedal mammals (29 dogs and 32 pigs) and 28 primates (Macaca apes). Moreover, macroslices of Macaca ape and of 182 human fetuses were examined histologically. RESULTS: In quadrupedal mammals, we found no traces of any adipose masses comparable to the human mesorectum nor were there ligaments of suspension. In the ape, the adipose tissue in the mesosigmoid forms an adipose cuff that completely surrounds the extraperitoneal rectum. Two dense connective bands were found between the lateral wall of the pelvis and the perirectal tissue. Both the mesorectum and the lateral ligaments were clearly identified in the sections of human fetus only at the end of the fifth month but not earlier. CONCLUSIONS: On the basis of our analysis of 3 animal species, we conclude that the mesorectum and lateral ligaments are absent in quadrupedal mammals but are present in primates. Therefore, we hypothesize that these structures appeared with the attainment of the upright position, even though other hypotheses are possible. PMID- 17115316 TI - Laparoscopic rectovaginopexy for rectal prolapse. AB - BACKGROUND: Open rectovaginopexy is an effective procedure for the treatment of both rectal prolapse and anterior rectocele. This study investigates our results of laparoscopic rectovaginopexy (LRVP). METHODS: A consecutive series of 14 patients (median age, 73 years; range 24-92) with rectal prolapse was planned for LRVP. Pre-, per- and postoperative parameters were recorded. Followup was performed at the outpatients' clinic. RESULTS: The median length of hospital stay was 6 days (range, 3-14). There was one fatal cerebrovascular accident 14 days postoperatively; this patient was excluded from further analysis. Median follow up was 7 months (range, 0.75-38). During follow-up, 11 of 13 patients (85%) experienced resolution or major improvement of their symptoms. Anal incontinence was diminished in 9 of 13 cases (69%). Constipation improved in 2 of 3 patients (66%). These three patients experienced a combination of both anal incontinence and costipation, preoperatively. Recurrence occurred in 2 patients (15%). Two others had a minor residual mucosal prolapse. No patients reported symptoms suggestive of operation-induced constipation or dyspareunia. CONCLUSIONS: LRVP is feasible, and seems to be an effective procedure for rectal prolapse. No operation-induced constipation was observed in this series. Taking into account the age and co-morbidities of these patients, morbidity and mortality may be considered acceptable. PMID- 17115317 TI - Early results of the treatment of internal hemorrhoid disease by infrared coagulation and elastic banding: a prospective randomized cross-over trial. AB - BACKGROUND: Rubber band ligation (RBL) is probably the most commonly performed nonsurgical therapy for hemorrhoidal disease. Infrared coagulation (IRC) is one of the most recent advances based on the use of "heat". Recent studies have demonstrated similar efficacy for both modalities. This prospective randomized crossover trial compared IRC and RBL for pain, complications, effectiveness, and patient satisfaction and preference in the treatment of internal hemorrhoids (IH). METHODS: Patients were randomized to receive either RBL (Group A) or IRC (Group B) for treatment of the first hemorrhoid; in a second procedure two weeks later, patients underwent the other procedure on the second hemorrhoid, thereby serving as their own control. The procedure preferred by the patient was employed two weeks later for the third hemorrhoid. Post-treatment pain was evaluated on a visual analog scale and on the basis of the percentage of patients requiring analgesics. Bleeding and early outcome of treatment were also recorded, together with the patient's satisfaction. RESULTS: A total of 94 patients were included in this study (47 patients in each group). At 30 minutes and 6 hours after treatment, pain scores were significantly higher in patients treated with RBL than in those treated with IRC (p<0.01). There was no significant difference in pain scores between the two procedures immediately and 24 hours after the procedures (p<0.05). After 72 hours and one week, the pain scores for RBL and IRC were similar. The percentage of patients using analgesics was significantly higher in RBL group than in IRC group at 6 hours (29.6% vs. 19.2%, respectively; p<0.05) and 24 hours (22.5% vs. 13.5%, respectively; p<0.05) after treatment. However, significant differences were not noted at 72 hours (12.7% vs. 6.4%; p<0.05) and one week (5.6% vs. 7.1%; p>0.05) after the procedures. There were significantly higher incidences of bleeding immediately, 6 hours, and 24 hours after RBL compared to IRC (immediate: 32.4% vs. 4.3%; 6 hours: 13.4% vs. 3.6%, 24 hours: 26.8% vs. 10.2%, respectively; p<0.01). However, there were no significant differences noted regarding the incidence of bleeding between the two groups at 72 hours. Complications were more likely after RBL than IRC, however this difference was not significant (p>0.05). Overall, 91 patients (96.8%) were successfully treated and 93 patients (99%) were very satisfied with the treatment. In the third treatment session, 50% of patients selected RBL and 50% chose IRC. CONCLUSIONS: Both RBL and IRC were well-accepted and highly efficacious methods for the treatment of IH; in addition, both procedures were associated with relatively minor complications. However, RBL was associated with more pain than IRC in the 24-hour postoperative period. PMID- 17115318 TI - The effect of polyethylene glycol and butyrate on anastomotic healing in the rat colon. AB - BACKGROUND: The use of mechanical bowel preparation is much debated. METHODS: We evaluated the effects of polyethylene glycol (PEG), with or without a single dose of 3.0 mmol butyrate (BUT), on the bursting pressure (BP) of an intact colon segment and a colon anastomosis in rats. Also, histopathologic damage was studied. RESULTS: In rats without colectomy, the mean BP was 159.2 mmHg (SD=18.9) after PEG treatment and 116.7 mmHg (SD=27.5) in controls (p=0.001). In rats with colectomy, the mean BP was 90.4 mmHg (SD=45.9) in the PEG group, 108.0 mmHg (SD=31.9) in the BUT group, and 102.7 mmHg (SD=44.7) in controls (p=0.44). No significant differences in histopathologic scores were observed between rats treated with PEG and controls. CONCLUSIONS: PEG does not interfere with anastomotic healing in rats as measured by BP. No benefit of a single dose of butyrate was observed. PMID- 17115319 TI - Minimally invasive resection for colorectal cancer: perioperative and medium-term results in an unselected patient group at a single institution. AB - BACKGROUND: The use of laparoscopy for colorectal cancer resection is still controversial. METHODS: We prospectively analyzed the outcome of minimally invasive resection for colorectal cancer, performed at our institution from 1998, when laparoscopic surgery became the treatment of choice for colorectal cancer, until 2004. All patients undergoing elective resection were assessed in terms of perioperative results (duration of surgery, number of lymph nodes removed, length of specimen, rate of conversion, complications) and survival. Patients were assessed yearly with follow-up visits and telephone interviews. RESULTS: In the study period, 302 patients (mean age 66.1 years; range, 32-93 years) underwent 114 left hemicolectomies, 108 low anterior resections, 61 right hemicolectomies, 12 Miles procedures, 4 subtotal colectomies, and 3 transverse colon resections. Surgery took an average of 226 minutes (SD=71 min). The number of lymph nodes removed was 14+/-8. The conversion rate was 10%; most of the conversions were due to locally advanced cancer (15 cases) and bowel distension (7 cases). Fifteen anastomotic leaks were observed (5%). Twenty patients needed reoperation and two died: one of septic shock due to an anastomotic leak; the other of electrolyte imbalance and dehydration after peritonitis due to a bowel loop injury. Follow-up was available for 91% of patients. Cancer-related survival curves showed a 90% survival for stage II, 85% for stage III, and 10% for stage IV disease, 30 months after surgery. CONCLUSIONS: Minimally invasive laparoscopic resection for colorectal cancer enables an oncologically adequate resection with complication and survival rates that are no worse than are to be expected after traditional open surgery. Locally advanced tumor and bowel distension are the most frequent reasons for conversion to open surgery. PMID- 17115320 TI - Transperineal versus hydrogen peroxide-enhanced endoanal ultrasonography in never operated and recurrent cryptogenic fistula-in-ano: a pilot study. AB - BACKGROUND: Accurate delineation of anal fistula anatomy in recurrent cases will assist in surgical fistula eradication whilst preserving continence. Recently, transperineal ultrasonography (TPUS) has been used in perirectal inflammation where there may be advantage over endoanal ultrasonography (EAUS) in complex fistulae- in-ano which lie outside the focal range of the endoanal probe. We assessed the sensitivity of these two imaging modalities to characterize fistula in-ano, compared to surgical findings. METHODS: Hand-held 7.5 MHz TPUS was performed in the axial and sagittal planes in never-operated (Group 1, n=10) and recurrent (Group 2, n=10) cryptogenic fistulae where the ultrasonographer was blinded to the initial operative findings. This was compared with hydrogen peroxide-enhanced EAUS using a 7.5 MHz rotating probe, assessing the fistula anatomy, site of the internal opening, confirmation of Goodsall's rule and the presence of secondary tracks, abscess collections and significant horsehoeing of the track. RESULTS: Overall sensitivity for the detection of trans-sphincteric and extrasphincteric fistulae was 100% using both techniques with a 90% sensitivity for TPUS and an 85% sensitivity for EAUS in the prediction of the internal fistula opening site. The TPUS sensitivity for horseshoeing was poor (28.6%) as was the detection of ancillary abscesses confirmed at surgery (63.6%) but TPUS demonstrated rectovaginal fistulae. CONCLUSION: TPUS is a novel technique for use in perirectal infection which has a significant learning curve but which is highly accurate for prediction of the anatomy of complex recurrent as well as simple anal fistulae. PMID- 17115321 TI - Different segmental transit times in patients with irritable bowel syndrome and "normal" colonic transit time: is there a correlation with symptoms? AB - BACKGROUND: The Rome criteria serve as gold standard for establishing a diagnosis of irritable bowel syndrome (IBS), but only represent a cluster of symptoms. On the other hand, measurement of colonic transit time (CTT) with radiopaque markers is a solid and more objective method to quantify functional abnormalities. The goal of this study was to investigate whether the IBS symptoms, as defined in the Rome II criteria, correspond to objective physiological parameters, i.e. CCTs. METHODS: The study enrolled 148 healthy control subjects and 1385 consecutive IBS patients. Transit times were measured for the whole rectocolon (overall CTT) and for 3 segments (right colon, left colon, rectosigmoid area); segmental distribution of markers and diffusion coefficients were also assessed. In order to analyze homogeneous groups, we restricted analysis to subjects with "normal" CTT (< or =70 hours). RESULTS: Six hundred forty four IBS patients (46%) and 14 control subjects (9%) had CTT >70 h and were eliminated. In subjects with CTT < or =70 h, CTT did not follow a normal (Gaussian) distribution. We identified 3 different CTT clusters in healthy controls and 4 clusters in IBS patients. Even if CTT was not significantly different between clusters, each cluster was characterized by a specific pattern of segmental colonic transit. There was a marked gender difference: women had longer overall CTT values than men, both in control and IBS patient groups (p<0.001). However, female IBS patients had significantly shorter colorectal transit times than female controls (p<0.001), as well as faster transit than in men through the left colon and rectosigmoid area. There were no significant differences in transit time between male IBS patients and male controls with the exception of a faster rectal transit in IBS patients (p<0.01). There was no association between segmental colonic transit values and sign or symptoms comprising the Rome II criteria. CONCLUSIONS: In subjects with CTT < or =70 h, CTT does not follow a normal distribution but is clustered in subgroups that can be distinguished only by measuring segmental colonic transit. Within these subgroups, there is a marked difference in transit times between IBS patients and normal subjects, suggesting that IBS patients with "normal" CTT are not "normal". The Rome II criteria do not reflect differences in segmental transit times in IBS patients with "normal" CTT. We therefore propose to evaluate segmental transit times in IBS patients with "normal" CTT, before and after treatment, in order to correctly interpretate variations in signs and symptoms. These findings have important implications in evaluating the effect of drugs on bowel function and should help define better inclusion criteria for studies evaluating new drugs for the treatment of IBS. PMID- 17115322 TI - Outcome of parastomal hernia repair with and without midline laparotomy. AB - BACKGROUND: Surgical options for parastomal hernia (PSH) repair are primary fascial, mesh repair, and relocation with or without midline laparotomy. Overall, recurrence rates are higher after fascial repairs than after relocation. However, stoma relocation may require a midline laparotomy with higher associated morbidity. The aim of this study was to assess the outcome of PSH repair with relocation with or without a midline laparotomy. METHODS: All patients who underwent PSH repair with relocation were identified from a clinical database. Data were collected by retrospective review of medical records including patient demographics, presenting symptoms, predisposing factors, type of surgery, postoperative complications, recurrence, and follow-up. Patients were divided into two subgroups, with or without a midline laparotomy. In patients without a laparotomy, the stoma was intraperitoneally mobilized, passed behind the abdominal wall, and delivered and matured through a premarked stoma site, across the midline. RESULTS: Between 1992 and 2001, a total of 27 patients underwent PSH repair with relocation of the stoma to the opposite side of the abdominal wall. Of these, the operation was performed without a midline laparotomy in 11 patients (41%). There were no significant differences in age, gender, body mass index, and the duration of hernia between the non-laparotomy and laparotomy groups. Prior abdominal surgery was recorded for 3 patients in the group without a laparotomy and for 9 patients in the group with a laparotomy (p=NS). Although not quantified, patients in the non-laparotomy group were less likely to have significant intraabdominal adhesions. Conversely, patients in the laparotomy group had more advanced adhesions. The operative time was longer in the group with a laparotomy than in the group without [96.8 (50-220) minutes vs. 123.9 (45 360) minutes; (p=NS)], and the mean hospital stay was significantly less in patients without vs. with a laparotomy [5.5 (SD=1.6) days vs. 9.5 (SD=3.8) days, respectively; (p<0.05)]. There was only one recurrence in the group without a laparotomy compared to 3 in the group with a laparotomy. The mean follow-up periods were 36.8 and 56.6 months in the groups without and with a laparotomy, respectively. The postoperative complications included wound infection that occurred in 3 patients in each group. CONCLUSIONS: PSH repair with relocation without laparotomy was associated with a significantly shorter hospital stay, possibly due to the lack of a midline abdominal wound. It may not be feasible in patients with significant intraabdominal adhesions. PMID- 17115323 TI - High prevalence of smoking among urban-dwelling Canadian men who have sex with men. AB - A small but consistent literature from the United States suggests increased risk for smoking among lesbians and men who have sex with men (MSM). Few studies have investigated smoking among MSM in other countries where different social norms and restrictions on smoking in public apply. We measured smoking behaviours in a convenience sample of urban-dwelling young Canadian MSM (median age 28 years). We compared the prevalence of smoking among MSM with that among other men in British Columbia (BC) using National Population Health Survey data to compute an age adjusted standardized prevalence ratio (SPR). Independent predictors of smoking among MSM were identified using adjusted odds ratios (OR) with 95% confidence intervals (CI). Smoking during the previous year was reported by twice as many MSM (54.5% of 354) as other men in BC (25.9%) (SPR = 1.94, 95% CI 1.48-2.59), with largest differentials observed among men under 25 years of age. In multivariable analyses, smoking among MSM was significantly associated with younger age (OR 0.94, CI 0.88-1.00 per year), greater number of depressive symptoms (OR 1.12, CI 1.06-1.19 per symptom) and Canadian Aboriginal ethnicity (OR 2.64, CI 1.05-6.60). In summary, MSM in our study were twice as likely to smoke as other men in BC; the greatest differences were observed among the youngest men. The design of effective prevention and cessation programs for MSM will require identification of the age-dependent determinants of smoking initiation, persistence, and attempts to quit. PMID- 17115324 TI - Dentin alteration of deciduous teeth in human hypophosphatemic rickets. AB - Familial hypophosphatemic rickets is in most cases transmitted as an X-linked dominant trait and results from mutation of the PHEX gene, predominantly expressed in osteoblast and odontoblast. Patients have been reported to display important dentin defects, and therefore, we explored the dentin structure, composition, and distribution of extracellular matrix (ECM) molecules in hypophosphatemic human deciduous teeth. Compared to age-matched controls, the dentin from hypophosphatemic patients exhibited major differences: presence of large interglobular spaces resulting from the lack of fusion of calcospherites in the circumpulpal dentin; defective mineralization in the interglobular spaces contrasting with normal Ca-P levels in the calcospherites on X-ray microanalysis; abnormal presence of low-molecular weight protein complexes recognized on Western blots by antibodies against matrix extracellular phosphoglycoprotein (MEPE), dentin sialoprotein, osteopontin, and reduced osteocalcin (OC) level; and accumulation in the interglobular spaces of immunolabeling with antibodies against DSP, dentin matrix protein, bone sialoprotein, MEPE and OC, while chondroitin/dermatan sulfate glycosaminoglycans were exclusively located inside calcospherites. Alterations of the post-translational processing or partial degradation of some ECM appear as key factors in the formation of the defective hypophosphatemic dentin. PMID- 17115325 TI - The prognostic impact of successful cardioversion of atrial fibrillation in patients with organic heart disease. AB - The aim of the study was to evaluate the prognostic impact of successful cardioversion (CV) compared to failed CV in patients with atrial fibrillation (AF) and organic heart disease. A total of 471 consecutive patients with organic heart disease from the prospective single center anticoagulation registry ANTIK who underwent CV of AF or atrial flutter were analyzed. 417 patients (89%) could be successfully cardioverted. In 54 patients (11%) CV failed, these patients remained in AF. After 5 years there were 92 (24%) deaths among patients with restored sinus rhythm at index admission and 20 (38%) deaths among those who remained in AF after CV (unadjusted OR 1.9, 95% CI 1.1-3.6). After adjustment for age, gender and ejection fraction, successful CV was not associated with a beneficial effect on mortality (OR 0.72, 95% CI 0.43-1.21). Thus, successful CV is not an independent predictor of mortality on multivariate analysis. However, it remains a marker for a better prognosis in patients with organic heart disease as these patients have a lower unadjusted longterm mortality. PMID- 17115326 TI - Atrial fibrillation in carcinoid heart disease: The role of serotonin. A review of the literature. PMID- 17115328 TI - Chromatin domains and the interchromatin compartment form structurally defined and functionally interacting nuclear networks. AB - In spite of strong evidence that the nucleus is a highly organized organelle, a consensus on basic principles of the global nuclear architecture has not so far been achieved. The chromosome territory-interchromatin compartment (CT-IC) model postulates an IC which expands between chromatin domains both in the interior and the periphery of CT. Other models, however, dispute the existence of the IC and claim that numerous chromatin loops expand between and within CTs. The present study was undertaken to resolve these conflicting views. (1) We demonstrate that most chromatin exists in the form of higher-order chromatin domains with a compaction level at least 10 times above the level of extended 30 nm chromatin fibers. A similar compaction level was obtained in a detailed analysis of a particularly gene-dense chromosome region on HSA 11, which often expanded from its CT as a finger-like chromatin protrusion. (2) We further applied an approach which allows the experimental manipulation of both chromatin condensation and the width of IC channels in a fully reversible manner. These experiments, together with electron microscopic observations, demonstrate the existence of the IC as a dynamic, structurally distinct nuclear compartment, which is functionally linked with the chromatin compartment. PMID- 17115329 TI - CENP-B box and pJalpha sequence distribution in human alpha satellite higher order repeats (HOR). AB - Using our Key String Algorithm (KSA) to analyze Build 35.1 assembly we determined consensus alpha satellite higher-order repeats (HOR) and consensus distributions of CENP-B box and pJalpha motif in human chromosomes 1, 4, 5, 7, 8, 10, 11, 17, 19, and X. We determined new suprachromosomal family (SF) assignments: SF5 for 13mer (2211 bp), SF5 for 13mer (2214 bp), SF2 for 11mer (1869 bp), SF1 for 18mer (3058 bp), SF3 for 12mer (2047 bp), SF3 for 14mer (2379 bp), and SF5 for 17mer (2896 bp) in chromosomes 4, 5, 8, 10, 11, 17, and 19, respectively. In chromosome 5 we identified SF5 13mer without any CENP-B box and pJalpha motif, highly homologous (96%) to 13mer in chromosome 19. Additionally, in chromosome 19 we identified new SF5 17mer with one CENP-B box and pJalpha motif, aligned to 13mer by deleting four monomers. In chromosome 11 we identified SF3 12mer, homologous to 12mer in chromosome X. In chromosome 10 we identified new SF1 18mer with eight CENP-B boxes in every other monomer (except one). In chromosome 4 we identified new SF5 13mer with CENP-B box in three consecutive monomers. We found four exceptions to the rule that CENP-B box belongs to type B and pJalpha motif to type A monomers. PMID- 17115330 TI - Identification and physical mapping of induced translocation breakpoints involving chromosome 1R in rye. AB - To obtain translocations involving specific chromosomes in rye, pollen of a line in which chromosome 1R has large C-bands on its two telomeres, but which lacks C bands (or has very small ones) on the telomeres of the remaining chromosomes, was X-irradiated. All translocations involving the labelled chromosome (1R) could be easily recognized in C-banded mitotic metaphases. The non-labelled chromosome involved in each translocation was identified either from mitotic C-banding analysis or from the meiotic configurations observed in some specific progenies. A physical map including 40 translocation breakpoints has been developed by means of synaptonemal complex (SC) analysis of well-paired pachytene quadrivalents. The results agree with the hypothesis of chromosomes 2R to 7R having similar probabilities of participating in translocations with chromosome 1R. However, the locations of the breakpoints are not entirely random: an excess of translocation breakpoints located on the short arm of chromosome 1R was obtained, and the two acentric translocated segments of each translocation show a trend towards having similar sizes. The possible reasons for these two non-random situations are discussed. PMID- 17115331 TI - Triploid origin of the gibel carp as revealed by 5S rDNA localization and chromosome painting. AB - 5S ribosomal DNA (rDNA) was isolated and sequenced from the gibel carp Carassius auratus gibelio with 162 chromosomes and crucian carp Carassius auratus with 100 chromosomes, and fluorescent probes for chromosome localization were prepared to ascertain the ploidy origin and evolutionary relationship between the two species. Using fluorescence in-situ hybridization (FISH), major 5S rDNA signals were localized to the short arms of three subtelocentric chromosomes in the gibel carp and to the short arms of two subtelocentrics in the crucian carp. In addition, some minor signals were detected on other chromosomes of both species. Simultaneously, six chromosomes were microdissected from the gibel carp metaphase spreads using glass needles, and the isolated chromosomes were amplified in vitro by degenerate oligonucleotide primed-polymerase chain reaction (DOP-PCR). Significantly, when the DOP-PCR-generated probes prepared from each single chromosome were hybridized, three same-sized chromosomes were painted in each gibel carp metaphase, whereas only two painted chromosomes were observed in each crucian carp metaphase spread. The data indicate that gibel carp is of triploid origin in comparison with diploid crucian carp. PMID- 17115332 TI - On the positions of centromeres in chicken lampbrush chromosomes. AB - Using immunostaining with antibodies against cohesin subunits, we show here that cohesin-enriched structures analogous to the so-called centromere protein bodies (PB) are the characteristic of galliform lampbrush chromosomes. Their centromeric location was verified by FISH with certain DNA probes. PB-like structures were used as markers for centromere localization in chicken lampbrush chromosomes. The gap predicted to be centromeric in current chicken chromosome 3 sequence assembly was found to correspond to the non-centromeric cluster of CNM repeat on the q-arm of chromosome 3; the centromere is proposed to be placed at another position. The majority of chicken microchromosomes were found to be acrocentric, in contrast to Japanese quail microchromosomes which are biarmed. Centromere cohesin-enriched structures on chicken and quail lampbrush microchromosomes co-localize with pericentromeric CNM and BglII- repeats respectively. FISH to the nascent transcripts on chicken lampbrush chromosomes revealed numerous non-centromeric CNM clusters in addition to pericentromeric arrays. Complementary CNM transcripts from both C- and G-rich DNA strands were revealed during the lampbrush stage. PMID- 17115333 TI - Extracolonic tumors of the gastrointestinal tract detected incidentally at screening CT colonography. AB - PURPOSE: The aim of this article is to report our experience with incidental detection of extracolonic tumors of the gastrointestinal tract identified prospectively at screening CT colonography. METHODS: A total of 2014 patients (1097 females, 917 males; mean age, 56.9 years) underwent primary CT colonography evaluation at our institution over an 18-month period. Following cathartic preparation and colonic distention, supine and prone multidetector CT scans were obtained with thin-collimation low-dose technique without intravenous contrast. We reviewed our database for lesions of the extracolonic gastrointestinal tract that were detected during the prospective reading. RESULTS: Focal extracolonic gastrointestinal tract lesions were prospectively detected in 10 (0.5 percent) of 2014 patients (8 females; 2 males; mean age, 58.5 years). All patients were asymptomatic. Tumor locations included ileum (n = 3), stomach (n = 3), jejunum (n = 2), and appendix (n = 2). Mean tumor size was 2.2 (range, 0.8-3.4) cm. Lesions in eight patients were subsequently confirmed by conventional or capsule endoscopy and/or by intravenous contrast-enhanced CT. Seven lesions were surgically excised and one was removed at endoscopy; two patients with lipomas did not undergo further evaluation or treatment. Final diagnoses were benign in all cases and included lipoma (n = 3), small-bowel hamartoma (n = 2), appendiceal mucinous cystadenoma (n = 2), gastric leiomyoma (n = 1), small-bowel lymphangioma (n = 1), and gastric fundic gland polyp (n = 1). CONCLUSIONS: Incidental extracolonic tumors of the gastrointestinal tract detected at screening CT colonography were all asymptomatic and benign but often prompted more invasive workup. Although the incidence of these tumors was relatively low, widespread population screening with CT colonography would result in new surgical referrals for these findings. PMID- 17115334 TI - Functional results of laparoscopic resection rectopexy for symptomatic rectal intussusception. AB - PURPOSE: This study was designed to assess the role of laparoscopic resection rectopexy for symptomatic rectal intussusception in patients who failed medical treatment. The functional outcomes of laparoscopic resection rectopexy were evaluated. METHODS: Patients who underwent laparoscopic resection rectopexy for rectal intussusception between July 1998 and November 2004 were identified. All patients with obstructed defecation failing medical treatment were included. Data were prospectively collected for the perioperative period. A follow-up questionnaire was used to assess functional outcome. RESULTS: Between 1998 and 2004, a total of 56 patients (53 females (95 percent); age range, 23-83 years) underwent laparoscopic resection rectopexy for rectal intussusception. The median operative time was 123 minutes. Morbidity was 7 percent, and there was no mortality. Fifty-two patients were available for follow-up, and of these 33 (63 percent) reported an overall improvement in their function after surgery. Of 28 patients suffering constipation, 15 (53 percent) reported an improvement in bowel frequency. Sixty-seven percent of patients incontinent before surgery improved. Symptoms of incomplete evacuation resolved in 38 percent of affected patients. Thirty-six percent of patients needing to strain at stool did not have this problem after surgery. Median follow-up was 44 (range, 15-92) months. CONCLUSIONS: The management of patients with rectal intussusception and obstructed defecation failing medical treatment is challenging. Laparoscopic resection rectopexy is an option that might offer symptomatic relief and improved function. Further studies are required to define the selection criteria to optimize the outcome in this patient group. PMID- 17115336 TI - Risk factors for anemia in patients with ileal pouch-anal anastomosis. AB - PURPOSE: Anemia is frequently observed in patients with ileal pouch-anal anastomosis. The identification of the underlying causes can be challenging. This study was designed to define the prevalence and to identify etiologic factors for anemia in this patient population. METHODS: A prospectively maintained database and medical records of patients who had restorative proctocolectomy between 1998 and 2005 were reviewed. All patients with laboratory evaluation at least six months after the surgery were studied. The last reported hemoglobin served as the index value. All patients with anemia (hemoglobin < 13.5 g/dl for males, <12 g/dl for females) were identified. A second group of randomly selected, ileal-pouch patients with normal hemoglobin served as control. Demographic and clinical variables were evaluated. RESULTS: A total of 389 patients (214 males) had documented hemoglobin values. Sixty-seven patients (17 percent; 40 males) had anemia. The prevalence of anemia was 19 and 15 percent in males and females, respectively. The prevalence was 17 percent among patients with underlying ulcerative colitis vs. 26 percent in patients with familial adenomatous polyposis (P = 0.27). The mean hemoglobin in the anemia group was 11.4 (median, 11.7) g/dl. One patient (2 percent) had severe (<7 g/dl), 11 (16 percent) had moderate (7-9.9 g/dl), and 55 (82 percent) had mild (> or =10 g/dl) anemia. One patient (2 percent) had macrocytic, 16 (24 percent) had microcytic, and 49 (74 percent) had normocytic anemia. Sixteen patients (24 percent) had unidentified causes for anemia. Multivariable analysis showed that the presence of malignancy or desmoid tumor and the J-pouch configuration were the only independent risk factors associated with anemia. CONCLUSIONS: Anemia is common in ileal-pouch patients. Malignancy or desmoid tumor and J-pouch configuration are independent risk factors for anemia. One-fourth of the patients with anemia have unclear etiology. PMID- 17115337 TI - Lymphatic vessel density at the site of deepest penetration as a predictor of lymph node metastasis in submucosal colorectal cancer. AB - PURPOSE: Lymph node metastasis is an important factor that influences curability after endoscopic treatment of submucosal colorectal cancer. This study was designed to determine the usefulness of identification of lymphatic vessels by immunohistochemistry in predicting lymph node metastasis of submucosal colorectal cancer. METHODS: Lymphatic involvement was assessed by hematoxylin and eosin staining and podoplanin immunostaining on samples resected from 268 patients with submucosal colorectal cancer. Lymphatic vessel density was estimated by two investigators by average count of three fields (x200) in the area of greatest number of podoplanin-positive capillaries at the site of deepest submucosal penetration. Relations with other clinicopathologic parameters also were investigated. RESULTS: Lesions with high lymphatic vessel density (> or =9 vessels per field) showed a significantly greater incidence of lymph node metastasis than did those with low lymphatic vessel density (<9 vessels per field; 23.3 vs. 8.4 percent). By multivariate analysis, lymphatic vessel density was determined to be an independent risk factor for lymph node metastasis of submucosal colorectal cancer (P = 0.0044). Lymphatic vessel density also correlated with tumor budding and the degree of inflammation at the invasive front. CONCLUSIONS: Identification of lymphatic vessels by podoplanin immunostaining provides objective and accurate evaluation of lymphatic involvement. Lymphatic vessel density at the site of deepest penetration is a useful predictor of lymph node metastasis of submucosal colorectal cancer. PMID- 17115338 TI - Factors affecting circumferential resection margin involvement after rectal cancer excision. AB - PURPOSE: This study was designed to assess factors affecting rates of circumferential resection margin involvement after rectal cancer excision, the association between circumferential resection margin involvement rates for patients undergoing anterior resection and abdominoperineal excision within the same unit, and trends in outcomes between units. METHODS: Data about patients undergoing rectal cancer excision between 2000 and 2003 were extracted from the Association of Coloproctology of Great Britain and Ireland database. Multivariate logistic regression analysis was used to identify independent predictors of circumferential resection margin involvement. Pearson correlation coefficient was used to evaluate the association between circumferential resection margin involvement for anterior resection and abdominoperineal excision. RESULTS: A total of 1,430 patients satisfied the inclusion criteria. The circumferential resection margin involvement rate for anterior resection (n=794) was 6.7 percent, between hospital variability was 0 to 40 percent, and for abdominoperineal excision (n=521) was 17.6 percent, between hospital variability 0 to 100 percent. Independent predictors of circumferential resection margin involvement were T stage (P<0.001), nodal involvement (P=0.007), and operative procedure (P<0.001). Units with a high circumferential resection margin involvement rate for anterior resection also had a high circumferential resection margin involvement rate for abdominoperineal excision (Pearson correlation=0.349; P=0.01). CONCLUSIONS: Circumferential resection margin involvement is more common in lymph-node positive tumors and is more common after abdominoperineal excision compared with anterior resection. This relationship was consistent across units irrespective of their individual circumferential resection margin involvement rates. PMID- 17115339 TI - Early results of surgery in 123 patients with pseudomyxoma peritonei from a perforated appendiceal neoplasm. AB - PURPOSE: Epithelial appendiceal tumors are uncommon but can present as an emergency simulating appendicitis, or unexpectedly at laparotomy, laparoscopy, or on cross-sectional imaging. Occult rupture with features of pseudomyxoma peritonei may be encountered. We report the operative findings, pathologic assessment, and early outcomes in 123 consecutive patients with a perforated appendiceal neoplasm presenting as pseudomyxoma peritonei. METHODS: From March 1994 to March 2004, 292 patients were referred to a peritoneal malignancy surgical treatment center. Complete tumor removal (cytoreduction) was attempted in selected patients and, if achieved, surgery was combined with intraoperative, intraperitoneal mitomycin C (10 mg/m(2)). RESULTS: In total, 123 patients (52 males; 41 percent) underwent laparotomy for a perforated appendiceal malignancy presenting as pseudomyxoma peritonei. The median age was 52 (range 30-77) years. Complete cytoreduction was achieved in 83 of 123 patients (67 percent), major palliative resection in 34 patients (28 percent), and 6 patients (5 percent) were inoperable. Postoperative mortality was 6 of 123 patients (5 percent). Kaplan Meier analysis of the 83 patients who had complete tumor removal predicted 75 percent disease-free survival at five years. CONCLUSIONS: A perforated appendiceal epithelial tumor most frequently presents as pseudomyxoma peritonei. This treatment strategy, involving surgery and intraperitoneal chemotherapy, can result in good outcomes in this rare and otherwise fatal disease. PMID- 17115340 TI - The role of positron emission tomography in the management of recurrent colorectal cancer: a review. AB - PURPOSE: Surgery remains the only option for potential cure in patients with recurrent colorectal cancer. Accurate staging modalities aid in the avoidance of futile surgery, which may result in considerable morbidity in patients with incurable disease. Current imaging techniques used in disease staging often are not sensitive enough to identify low-volume metastatic disease. This study reviews the role of positron emission tomography in the assessment of patients with suspected recurrent colorectal cancer. METHODS: A literature search using the PubMed, MEDLINE, and Embase database was performed, locating English language articles on positron emission tomography, positron emission tomography, recurrent colon, and/or rectal cancer. The references of these papers were searched manually for further references. RESULTS: Positron emission tomography is more sensitive and more specific than conventional diagnostic imaging for metastatic disease and local recurrence respectively. Studies confirm the superior ability of positron emission tomography scans compared with conventional diagnostic imaging in differentiating between scar tissue and invasive tumor. Positron emission tomography scanning is more sensitive and specific for the assessment of liver metastases (and probably in patients with lung metastasis) than conventional diagnostic imaging. Positron emission tomography is superior to conventional diagnostic imaging in the investigation of raised carcinoembryonic antigen in the postoperative patient and alters management in approximately 37 percent of patients with recurrent colorectal cancer. The limitations and cost effectiveness of positron emission tomography are discussed. CONCLUSIONS: Positron emission tomography scanning is emerging as the imaging modality of choice for patients being considered for surgery for locally recurrent colorectal cancer. Positron emission tomography has the greatest impact by detecting unresectable disease and thereby averting inappropriate surgery. Despite the high set-up costs, its use seems to be cost effective. PMID- 17115341 TI - Malone antegrade continent enema: an alternative to resection in severe defecation disorders. AB - PURPOSE: This study was designed to evaluate patient self-reported outcome of the Malone antegrade continent enema at a single institution in patients suffering from severe defecatory disorders. METHODS: A total of 18 patients (15 females; median age, 31 (range, 12-63) years) underwent a Malone antegrade continent enema (August 1999 to September 2004). The Malone antegrade continent enema technique has been previously described; however, in this series emphasis was placed on method appendix tunneling. Patients' charts were reviewed and follow-up telephone interviews were conducted. Indications for Malone antegrade continent enema were chronic constipation (n = 12), intractable fecal incontinence (n = 5), or both (n = 1). The underlying pathology included neurogenic (n = 2), congenital (n = 4), postsurgery-related (n = 4), irritable bowel syndrome (n = 6), and megarectum (n = 2). The appendix (n = 17) or cecum (n = 1) was used as a conduit. RESULTS: The mean follow-up was 18.5 (range, 3-67) months. Fourteen patients (78 percent) still use the Malone antegrade continent enema routinely and report good functional outcome. Three patients (20 percent) required stoma creation as subsequent alternate treatment. A total of 10 patients experienced 12 complications: 3 perioperative (infections) and 9 postoperative Malone antegrade continent enema use/nonuse complications (4 stomal orifice strictures, 2 fecal impactions, 2 appendiceal perforations, and 1 irrigation catheter knot). No patient experienced leakage from the appendiceal stoma. During the follow-up interval, one patient underwent proctectomy for megarectum. No failures occurred in patients with congenital or neurogenic disorders. CONCLUSIONS: Malone antegrade continent enema is a reasonable option for the treatment of select patients with severe defecation disorders. Good functional patient self-reported outcome was achieved by 78 percent of patients. The social inconvenience of stoma leakage is avoided with appropriate surgical technique. Malone antegrade continent enema is one option that provides a less invasive surgical alternative than colectomy or ileostomy for severe defecation disorders. PMID- 17115342 TI - The effect of capecitabine on the healing of colonic anastomoses in rats. AB - PURPOSE: Capecitabine is a fluoropyrimidine carbamate with antineoplasmatic activity, recommended for the treatment of colorectal cancer. This study was designed to assess the effectiveness of the perioperative administration of capecitabine on the healing process of colonic anastomosis. METHODS: Sixty Wistar rats were used, which were randomized in 2 groups of 30 each. The study group was subjected to colonic anastomosis and treated with therapeutic doses of capecitabine (359 mg/kg, or 2/3 of the mean toxic dose) by mouth one week before anastomosis and throughout the study. The control group received only placebo medication. Both groups were further divided into three subgroups, each of ten animals. In both study and control groups, ten animals were killed in each session on postoperative Days 3, 7, and 14. RESULTS: We found no negative impact on the healing of colonic anastomosis on capecitabine administration. The rate of anastomotic leakage and septic complications were not found to be significantly different between the study and control groups. The median bursting pressure was found to be significantly higher in the study subgroup killed on the third day (68 vs. 46 mmHg of the control group). CONCLUSIONS: Perioperative administration of capecitabine does not have a negative impact on colonic anastomosis in rats. PMID- 17115344 TI - Pharmacotherapy of obesity. AB - Obesity has become a significant health problem in industrialised and developing countries, and despite all nutritional and behavioural approaches, its prevalence is still increasing. In recent years, the identification and characterisation of central and peripheral mechanisms involved in the regulation of energy balance has made remarkable progress and provided numerous targets for novel anti-obesity agents. However, only few anti-obesity drugs are on the market and not many compounds have entered clinical development. In the present review, the clinically available agents are discussed and their pharmacological profiles are compared. Some of the drugs that are currently in clinical development are mentioned as examples of the possible future range of anti-obesity agents. Selected topics in drug discovery are presented to illustrate novel targets and concepts for the pharmacotherapy of obesity. PMID- 17115345 TI - Evaluation of the diagnostic value of the first thyroglobulin determination in detecting metastases after differentiated thyroid carcinoma surgery. AB - AIM: Evaluation of the diagnostic value of the first thyroglobulin (Tg) level measurement, performed after thyroidectomy, before another treatment, as an early marker of either metastases or local recurrence of differentiated thyroid carcinoma (DTC). MATERIALS AND METHODS: Data of 178 patients (160 women, 18 men, 14-79 years) with DTC and without known interference in Tg assay were evaluated retrospectively. In all patients, neck radioiodine uptake (Tup (24)), thyroid remnants volume (V), TSH and Tg were measured. The Tg/V and Tg/Tup (24) ratios were calculated to correct Tg concentration with regard to V and Tup (24). Six months after initial evaluation and routine therapy all patients underwent control examinations under endogenous TSH stimulation. RESULTS: During follow-up metastases or local recurrence were found in 32 patients. The groups of patients with no diagnosed metastases (M0) and with detected metastases (M1), did not differ with regard to V, serum TSH or Tup (24); difference between the two groups was found in Tg concentration (4.3 ng/ml VS 97.4 ng/ml; p=0.000001). The ratios of Tg/Tup (24) (p=0.000000) and Tg/V (p=0.004) were lower in the group M0 than M1. The areas under receiver operating characteristic curves (ROC) for Tg concentrations, Tg/Tup (24), and Tg/V ratios were 0.773 (95% CI - 0.655-0.892), 0.817 (0.709-0.925) and 0.712 (0.541-0.884), respectively. CONCLUSIONS: Both the absolute Tg concentration and Tg/V and Tg/Tup (24) ratios, determined after thyroidectomy but before another treatment in patients with metastases of DTC, diagnosed within 6 months after (131)I administration, are higher than those in patients without such metastases. This indicates that the mentioned parameters may be applied as early markers of either local recurrence or metastases of DTC. The highest discriminative value demonstrates Tg/Tup (24) ratio, Tg concentration has a lower value and Tg/V ratio has the lowest one. PMID- 17115346 TI - A cross-sectional study to investigate long-term cognitive function in people with treated pituitary Cushing's disease. AB - INTRODUCTION: It has been proposed that exposure to high levels of endogenous steroids in untreated pituitary Cushing's disease damages hippocampal structures leading to impairment in learning and memory processes. We hypothesised that patients with treated pituitary Cushing's disease would perform significantly worse on tests of cognitive ability than those with nonfunctioning pituitary adenomas. DESIGN: Sixteen adults with pituitary Cushing's disease (PCD) and 16 adults with non-functioning pituitary adenomas (NFA) undertook the following comprehensive neuropsychological assessments: National Adult Reading Test (NART: premorbid abilities), California Verbal Learning Test (CVLT 2 UK: learning and recall), Stroop (executive functioning), Trail-Making Test (TMT: executive functioning and attention), Adult Memory and Information Processing Battery (AMIPB: Information Processing Speed and Story Recall subtests). RESULTS: There was no significant difference in premorbid IQ scores (NFA mean=101 SD=13; PCD mean=102, SD=13), in verbal learning nor any significant difference in the percentage of verbal material retained in story recall (AMIPB). Performance on higher executive tasks Stroop and TMT and on measures of information processing was similar. However, there were significant decrements between some mean scores for both groups and published normative data with a clear association between higher HADS depression scores and impaired objective memory and attention which was not specific to PCD. CONCLUSIONS: We found no difference in cognitive function between patients with PCD and NFA. The results suggest a discrepancy between patients' subjective perception of functional cognitive impairments and objective findings on psychometric testing and point to the influence of affective symptoms on cognitive performance, particularly in Cushing's disease. PMID- 17115347 TI - The German Acromegaly Registry: description of the database and initial results. AB - Patient registries are valuable tools to study long-term morbidity and mortality of rare diseases. Acromegaly is rare (incidence 3-4/mill/year, prevalence 40 70/mill; approx. 300 new patients/yr and up to 5700 patients in Germany). Diagnostic and therapeutic possibilities have considerably improved, but treatment results remain often unsatisfactory. The main cause is residual disease activity after surgery, most importantly due to invasive macroadenomas. The German Acromegaly Registry is an initiative of the Pituitary Study Group of the German Endocrine Society (DGE). Formally established in January 2003 by the Board of the DGE, long-term financial support is guaranteed by an unrestricted grant from Novartis Pharma GmbH to the DGE. The registry cooperates closely with the United Kingdom and the Austrian registries. The aim of the German Acromegaly Registry is to establish a database of sufficient epidemiological strength in order to (1) document co-morbidity and mortality, (2) provide data on diagnostic and therapeutic procedures/effectiveness, (3) enable comparison of procedures in different national centres, (4) provide information for patient support groups/interaction with health care providers, (5) enable comparison with other national registries within Europe. The registry has at present 82 participating centres, and 42 have included patients (20 university clinics, 8 non-university hospitals, 14 centres in private practice). The database aims to include all acromegalic patients in Germany who are cared for and treated at present. Up to December 2005 1543 patients have been entered in a retrospective manner. Data collection is by external monitoring by highly trained study nurses who visit the individual centres. Inclusion is planned to continue at a rate of 500 per year. Starting in 2005 centres are revisited every 3 years at a rate of 500 per year (prospective phase of the registry). Quality of the data has been validated by an independent monitoring team which demonstrated high data concordance. CONCLUSIONS: Initial results of the German Acromegaly Registry show that it was possible to include a large number of patients within 3 years into the registry. Data quality has been validated and shown to be satisfactory. Therefore, the registry will be a useful tool to study long-term morbidity and mortality in a large series of patients. PMID- 17115348 TI - Biochemical markers of bone turnover during pregnancy: a longitudinal study. AB - OBJECTIVE: The objective of this study was to prospectively investigate the effect of pregnancy on biochemical markers of bone turnover in healthy pregnant women. METHODS: During the course of our longitudinal study, biochemical markers of bone remodeling were measured in all three trimester of pregnancy (first trimester: 12.5+/-1.8 SD, second trimester: 21.6+/-1 SD, third trimester: 34.8+/ 1.6 SD weeks of gestation). Serum type I collagen C-telopeptides (CTX) and a crosslinked peptide of the carboxy-terminal telopeptide of type I collagen (ICTP) were used as markers of bone resorption. Bone alkaline phosphatase (BAP) and the N-terminal propeptides of type I collagen (PINP) were used as biochemical markers of bone formation. Blood samples for the analysis of all 4 biochemical markers according to each trimester of pregnancy were available in 49 patients. RESULTS: The main changes for all biochemical markers were seen between the second and the third trimester. According to the markers of bone resorption, both serum CTX and ICTP showed a significant increase from the first to the third and from the second to the third trimester (p<0.001; median percentage change: CTX=101.5% and ICTP=40%). Concerning markers of bone formation, PINP showed a significant decrease from the first to the second trimester (p=0.001) followed by a significant increase from the second to the third trimester (p<0.001, 63.8%) and an overall increase from the first to the third trimester (p<0.001). BAP also showed a significant increase from the second to the third trimester (p<0.001; 51.7%) and an overall increase from the first to the third trimester (p<0.001). CONCLUSION: Markers of bone resorption were significantly increased during pregnancy. In contrast to bone resorption, markers of bone formation showed an increase as well as a decrease during pregnancy indicating a state of high bone turnover. This might coincide with the change in bone mineral density that was observed in some, but not all, studies using "dual-energy x-ray absorptiometry" (DXA) as well as "quantitative ultrasonometry" (QUS). PMID- 17115349 TI - Biphasic insulin aspart given thrice daily is as efficacious as a basal-bolus insulin regimen with four daily injections: a randomised open-label parallel group four months comparison in patients with type 2 diabetes. AB - AIMS: To show that a thrice daily meal-time biphasic insulin aspart (BIAsp) treatment regimen is as efficacious as a 4 times daily basal-bolus regimen with human isophane insulin (NPH) and insulin aspart (IAsp). METHODS: A multinational, randomised, open-label parallel-group trial in 394 patients with type 2 diabetes on a once or twice daily insulin regimen. Patients were randomised 1:1 to BIAsp or IAsp+NPH for 16 weeks. The BIAsp group was treated according to individual needs using BMI as a surrogate index of insulin resistance. Subjects administered BIAsp 70 (BMI< or =30 kg/m (2)) or BIAsp 50 (BMI>30 kg/m (2)) with breakfast and lunch and BIAsp 30 with dinner. The IAsp+NPH group injected IAsp at meals and NPH at bedtime as basal insulin. HbAlc levels after 16 weeks were compared between treatments using a predefined non-inferiority criterion of 0.4%. The incidence of hypoglycaemic episodes and adverse events was evaluated. RESULTS: Mean HbAlc (+/ SD) decreased from 9.1+/-0.7% to 7.8+/-1.0% with both treatments. Glycaemic control provided by BIAsp was non-inferior to that obtained by the IAsp+NPH (intention to treat ITT) population: diff, HbAlc -0.05%; 95% CI (-0.24; 0.14); per protocol (PP) population: diff, HbAlc -0.03%; 95% CI (-0.23; 0.16). Similar improvements in glycaemic control in both groups were confirmed by self-measured 8-point plasma glucose (PG) profiles, average and fasting PG concentrations, and average prandial PG increments. The incidence of adverse events and hypoglycaemic episodes was similar in the two treatment groups. CONCLUSIONS: A thrice daily meal-time BIAsp regimen is a suitable alternative to an intensified insulin regimen in people with inadequately controlled type 2 diabetes mellitus, and requires fewer daily injections than a basal-bolus therapy without compromising efficacy and safety. PMID- 17115350 TI - Does rapid transition to insulin therapy in subjects with newly diagnosed type 2 diabetes mellitus benefit glycaemic control and diabetes-related complications? A German population-based study. AB - OBJECTIVE: The aim of this study was to assess whether earlier transition to insulin in subjects with newly diagnosed type 2 diabetes improves glycaemic control and reduces diabetes-related complications. METHODS: Subjects with newly diagnosed type 2 diabetes, and 2 or more recorded glycosylated haemoglobin (HbA1c) values, were identified from the Mediplus Germany database between June 1993 and May 2001. Subjects were stratified by treatment group: diet and exercise, sulfonylurea drugs, antihyperglycaemic drugs, insulin, or insulin plus sulfonylurea or antihyperglycaemic drugs. Treatment modifications were tracked over time and a rapid transition to insulin recorded if insulin was the initial therapy administered or the immediate treatment after diet and exercise. The area under the curve (AUC) for HbA1c was calculated and a linear regression model used to explain AUC as a function of rapid transitioning to insulin. A Cox proportional hazard model assessed the relationship between the time to first complication and AUC, rapid transition to insulin, and the number of treatment modifications. RESULTS: Of the 3136 subjects who met the study entry criteria, just 151 (4.8%) were initiated on insulin; after 5 years only 811 (25.9%) subjects had received insulin therapy. In the regression model explaining AUC, rapid transition to insulin significantly improved glycaemic control (-0.20, p=0.03). The Cox proportional hazard model demonstrated that the time to first complication was negatively related to AUC (-0.05, p<0.01) and the rapid use of insulin (-0.27, p<0.01), and positively related to the number of treatment modifications (0.07, p<0.01). CONCLUSIONS: Using actual real world clinical practice data, the present study found that the immediate use of insulin in patients with type 2 diabetes improved blood glucose control as measured by the AUC for HbA1c readings. This, in turn, reduced the risk of diabetes-related complications. In contrast, we observed that a stepwise transition treatment pattern (switching from diet and exercise to sulfonylureas and then perhaps to antihyperglycaemic agents and finally insulin) increased the risk of diabetes related complications. Greater effort is required to remove some of the barriers currently preventing earlier initiation of insulin therapy in patients with type 2 diabetes. PMID- 17115351 TI - Starting insulin therapy in type 2 diabetes: twice-daily biphasic insulin Aspart 30 plus metformin versus once-daily insulin glargine plus glimepiride. AB - AIM: To compare the efficacy and safety of two analog insulins as starting regimens in insulin-naive Type 2 diabetes patients. METHODS: In this randomized, open-label parallel study, twice-daily biphasic insulin aspart 30 (30% soluble and 70% protaminated insulin aspart; BIAsp 30) plus metformin (met) was compared with once-daily insulin glargine (glarg) plus glimepiride (glim) in 255 insulin naive patients (131 male; mean+/-SD age, 61.2+/-9.1 years). Mean baseline HbA (1c) (+/-SD) was 9.2+/-1.4% and 8.9+/-1.3% for BIAsp 30 plus met ( N=128) and glarg plus glim ( N=127), respectively ( P=0.0747). Primary endpoint was the difference in absolute change in HbA (1c) between groups after 26 weeks of treatment. RESULTS: HbA (1c) change was significantly greater in the BIAsp 30 plus met group than the glarg plus glim group (between-group difference: -0.5% (95% CI: -0.8; -0.2); P=0.0002). Mean prandial plasma glucose increment was significantly lower for BIAsp 30 plus met compared with glarg plus glim: 1.4+/ 1.4 mmol/l vs. 2.2+/-1.8 mmol/l; P=0.0002. During the maintenance phase (weeks 6 26), one major hypoglycemic episode occurred in each group; 20.3% and 9% of patients experienced minor hypoglycemic episodes in the BIAsp 30 plus met and glarg plus glim groups, respectively ( P=0.0124). At end-of-trial, mean daily insulin doses were 0.40 U/kg BIAsp 30 and 0.39 U/kg glarg. Glarg plus glim treated patients experienced significant weight gain of 1.5 kg (95% CI: 0.84; 2.19; P<0.0001). Weight change with BIAsp 30 plus met of +0.7 kg was not statistically significant (95% CI: -0.07; 1.42; P=0.0762). CONCLUSIONS: Starting insulin in Type 2 diabetes patients with twice-daily BIAsp 30 plus met can reduce HbA (1c) and mean prandial plasma glucose increment to a greater extent than once daily glarg plus glim. PMID- 17115352 TI - Wegener's granulomatosis presenting as diabetes insipidus. AB - Pituitary gland involvement in Wegener's granulomatosis (WG) occurs most commonly in the form of central diabetes insipidus (CDI). However, CDI as a presenting manifestation of WG is very rare. We report two such cases; one of them had multi organ involvement at presentation, while other developed it during follow-up. CDI was reversible following cytotoxic drug therapy in one of them. PMID- 17115353 TI - The effects of promoting colorectal cancer screening on screening utilisation: evaluation of the German Campaign "Aktiv gegen Darmkrebs" (Action against Colorectal Cancer). AB - BACKGROUND: Although colorectal cancer is the second leading cause of cancer death in Germany, screening rates remain low. It is unknown whether local approaches to promoting colorectal cancer screening are effective. This study evaluates the effectiveness of a regional awareness campaign consisting of public information sessions and leaflet distribution, upon the use of colorectal cancer screening by faecal occult blood test (FOBT) and colonoscopy. METHODS: Data on FOBT and colonoscopy were collected by doctors for 12 months following the campaign, including reason for and result of the testing. 37 % (n = 43) of all physicians and practitioners in two administrative districts (249 632 inhabitants) that were exposed to the campaign participated in the study. It was recorded whether the individuals requesting colorectal cancer screening had been prompted by the campaign, so the number and outcome of campaign-related tests was determined. RESULTS: 3398 individuals (male: 54.6 %, female: 45.4 %, age: 64.3 +/ 8.3 years) underwent 3551 screening tests (2446 FOBT, 819 colonoscopy, 133 both). 141 additional diagnostic colonoscopies were performed because of a positive result in the FOBT. Adenomas were detected in 279 individuals, malignancies in 11 individuals. 225 FOBT (8.7 %) and 176 colonoscopies (18.5 %) were performed due to the campaign; among these individuals, 49 had adenoma(s), 1 had carcinoma. The effect could be attributed both to leaflets and to information sessions. The proportion of campaign-related screening tests declined over time. CONCLUSION: The regional information campaign could reach target individuals and motivate them to utilise colorectal cancer screening. The combination of mass media and personal communicative elements seems useful. PMID- 17115354 TI - [Attitudes towards cadaveric organ donation--results from a representative survey of the German population]. AB - The increasing deficit of organs causes a drastic decline in the quality of life and survival of numerous patients in need of a transplantation. The purpose of this representative community study was to survey attitudes toward transplantation in the German population and to identify underlying determinants. Unlike previous surveys, fears and concerns were elicited based on a concrete case vignette. Among the 1,002 participants, 90 % were in favour of organ donation in general; 21 % reported having signed an organ donor card. Consent to organ donation was associated with younger age and higher social class; the same was true for the possession of an organ donor card. In the virtual decision situation, the majority (77 %) voted in favour of an organ donation based on saving lives, consolation for bereaved and the absence of disadvantages for the donor. Common (up to 50 %), however, were also fears and concerns regarding determination of the time of death, displacement of medical concern from the donor to the recipient of the organ, utilisation of organs for other purposes, or explantation before death. The knowledge of the determinants identified, of existing fears and concerns are helpful not only for informing the public, but also for the dialogue with the next of kin of potential donors. Here, it may be helpful not only to address arguments pro organ donation, but also to address potential fears and concerns. PMID- 17115355 TI - [Blue rubber-bleb nevus syndrome and therapeutic double balloon enteroscopy]. AB - This is the first report about the endoscopic removal of hemangiomas in the gastrointestinal tract using the double-balloon enteroscopic technique. We report on a 16-year-old female patient with a 10-year history of chronic anemia due to recurrent gastrointestinal bleeding. Besides permanent iron substitution, up to 3 blood transfusions per week are necessary. At birth a blue rubber-bleb nevus syndrome was diagnosed, with blue angiomatous lesions preferring her skin and digestive tract. In the 8 years before admittance numerous conventional endoscopic procedures and one intraoperative endoscopy with laser coagulation of many cavernous hemangiomas were performed. In our department the successful treatment of 150 hemangiomas with argon plasma coagulation or polypectomy in combination with double-balloon enteroscopy and conventional endoscopy was achieved without complications. PMID- 17115356 TI - [An unusual case of upper GI bleeding: esophageal cancer metastasis at a percutaneous endoscopic gastrostomy site--case report and review of the literature]. AB - We present the case of a patient with an esophageal squamous cell carcinoma, who was treated primarily by radiotherapy. Due to dysphagia, the patient received a percutaneous endoscopic gastrostomy (PEG) without any sign of tumour at that time. Five months later the patient presented with an upper GI bleeding from a gastric ulcer, which histologically turned out to be a metastasis of the previously diagnosed squamous cell carcinoma. So-called "implantation metastases" at the percutaneous endoscopic gastrostomy site are rare and most of the cases have been described in patients with head and neck tumours. Moreover, the presentation as an upper GI bleed is very uncommon and needs the attention of both endoscopists as well as gastrointestinal oncologists. Clinicopathological features of this case with a brief review of the literature are presented. PMID- 17115357 TI - A rare case of primary rectal rhabdomyosarcoma in an adult. AB - We report an extremely rare case (1:200 000) of primary rectal rhabdomyosarcoma (RMS) in an adult. In the literature, this case report is the first one dealing with an adult patient. The diagnosis was assessed by computed tomography (CT), magnetic resonance imaging (MRI), positron emission tomography (PET), endoscopy and histological examination. Neoadjuvant systemic chemotherapy with adriamycin and isofosfamide followed by anterior rectum resection was performed. This article should implicate RMS as a rare differential diagnosis of atypical rectal tumours in adults. PMID- 17115358 TI - [Colorectal carcinoma screening in patients with type 2 diabetes mellitus]. AB - Colorectal carcinoma is one of the most common tumour entities in Western countries. Colorectal carcinoma and type 2 diabetes mellitus share common risk factors. Recent epidemiological studies show an increased risk for colorectal carcinomas in patients with type 2 diabetes mellitus, even more pronounced at therapy with sulfonylureas or insulin. Moreover, a 3-fold risk increase for patients with insulin-dependent type 2 diabetes mellitus in comparison to the general population has been observed. The hyperinsulinaemia hypothesis is based on the premise that elevated plasma levels of insulin and free IGF-1 promote the proliferation of colon cells and lead to a survival benefit of transformed colon carcinoma cells. This is reflected by an altered tumour biology; in patients with type 2 diabetes, tumour progression is more rapid and tumour-associated mortality is increased. Colorectal carcinoma represents an entity that is well amenable to and can potentially be avoided by screening colonoscopy. Recommendations for colorectal carcinoma screening should employ the recent epidemiologic evidence. All patients with type 2 diabetes should be recommended to undergo colonoscopy before starting insulin therapy, and screening intervals should not exceed 5 years. This work provides a review of the evidence, and an algorithm is proposed for a modified screening in patients with type 2 diabetes. PMID- 17115359 TI - [Treatment and management of celiac disease]. AB - In most patients the clinical course of celiac disease is unproblematic after the diagnosis has been made and a strict gluten-free diet is established. However, in rare cases complications like refractory sprue or lymphoma can occur. Individual management is required since the clinical presentation of celiac disease can be very heterogeneous. For example, it is a matter of controversy if asymptomatic patients, who have the same typical histological changes in their small bowel like patients with symptomatic celiac disease, should adhere to a gluten-free diet. A major problem is the compliance and the unintentional intake of gluten. A 100 % gluten-free diet is not possible since most food components are contaminated with trace amounts of gluten. Fortunately most patients tolerate these contaminations. Furthermore, the threshold for gluten contamination can differ highly among patients. One central point in patient care is the monitoring of a gluten-free diet and the timely recognition of complications. Therefore, the role of antibodies and duodenal histology in monitoring the course of the disease will be discussed. PMID- 17115360 TI - [Liver-directed gene and cell therapy: future or present?]. PMID- 17115361 TI - [Pancreatic cancer screening: mission impossible?]. PMID- 17115362 TI - [Endoscopy in IBD. A Consensus Report of the IBD Working of the Austrian Association of Gastroenterology and Hepatology]. AB - Ileocolonoscopy including biopsies is the first line investigation in suspected inflammatory bowel disease (IBD). In up to 90 % of the cases ulcerative colitis and Crohn's disease are differentiated on endoscopic presentation. Standardised reporting of endoscopic results increases the validity and comparability of IBD findings. When there is a firm diagnosis of IBD, colonoscopy should only be performed for specific questions. An upper gastrointestinal endoscopy is only indicated in patients with upper gastrointestinal symptoms. Push and capsule endoscopy should also be limited to specific questions and situations. IBD with extended colitis is associated with an increased risk for colorectal cancer. Endoscopic surveillance with accurate biopsy sampling is a valuable tool for the prevention of colorectal cancer. PMID- 17115366 TI - New biology, old metaphors. PMID- 17115367 TI - Our mate animals. PMID- 17115368 TI - Expanding the cell theory. PMID- 17115369 TI - [The role of cholesterol in Alzheimer's neuro-pathogenesis]. AB - Alzheimer's Disease (AD) is the most common neurodegenerative disorder in western societies affecting up to 15 million individuals worldwide.It leads to death after a progressive memory deficit and cognitive impairment accompanied by the appearance of two pathological hallmarks in specific brain areas: neurofibrillary tangles and amyloid plaques. Cholesterol homeostasis may play a key role in AD pathogenesis and this is supported by the demonstration that cholesterol-rich membrane domain, so-called Rafts,are disorganized in affected brains. Retrospective clinical studies indicate that individuals chronically treated with cholesterol synthesis inhibitors,statins, are at lower risk of developing AD but current literature is conflicting with regard to the neuroprotective effects of statins on cognitive impairment. Before recommending statins for prevention and/or treatment of AD it is important to investigate more the role of cholesterol levels in neurodegenerative disorders. PMID- 17115370 TI - Relevance of plant lectins in human cell biology and immunology. AB - Protein-carbohydrate interactions are used for intercellular communication. Mammalian cells are known to bear a variety of glycoconjugates. Lectins, first discovered in plants, are proteins which can specifically bind carbohydrates. Given the high affinity of plant lectins for carbohydrates, they have always been important as molecular tools in the identification, purification and stimulation of specific glycoproteins on human cells. Lectins have provided important clues to the repertoire of carbohydrate structures in animal cells. The discovery of plant lectins gave a great impulse to modern glycobiology. They represent important biochemical reagents for numerous applications in the biomedical field and in research. Sequence determinations and structural characterization helped to understand the mechanism of action in many biologic systems. Plant lectins have been fundamental in human immunological studies because some of them are mitogenic/activating to lymphocytes. Understanding the molecular basis of lectin carbohydrate interactions and of the intracellular signalling evoked holds promise for the design of novel drugs for the treatment of infectious, inflammatory and malignant diseases. It may also be of help for the structural and functional investigation of glycoconjugates and their changes during physiological and pathological processes. PMID- 17115371 TI - On the unfitness of natural selection to explain sexual reproduction, and the difficulties that remain. AB - In its essence, the explanatory potential of the theory of natural selection is based on the iterative process of random production and variation, and subsequent non-random, directive selection. It is shown that within this explanatory framework, there is no place for the explanation of sexual reproduction. Thus in Darwinistic literature, sexual reproduction - one of nature's most salient characteristics - is often either assumed or ignored, but not explained. This fundamental and challenging gap within a complete naturalistic understanding of living beings calls for the need of a cybernetic account for sexual reproduction, meaning an understanding of the dynamic and creative potential of living beings to continuously and autonomously produce new organisms with unique and specific constellations. PMID- 17115372 TI - A theoretical framework for defining some concepts in evolution. AB - We present a theoretical framework for biological evolution with the intention of giving precise mathematical definitions of some concepts in evolutionary biology such as fitness, evolutionary pressure, specialization and natural selection. In this framework, such concepts are identified with well-known mathematical terms within the theory of dynamical systems. We also discuss some more general implications in evolution; for instance, the fact that our model naturally exhibits a frequency spectrum of the type 1/f for low frequencies of evolutionary events. PMID- 17115373 TI - The history of hesperopithecus: the human-ape link that turned out to be a pig. AB - Nebraska Man was a fossil discovery that was regarded by several leading experts as important in understanding evolutionary history. The only evidence for this anthropod was a single tooth (which turned out to be a pigs' tooth). The discovery and controversy surrounding the Nebraska Man (Hesperopithecus haroldcookii hominoidea) fossil find and its importance in history are reviewed. Its supporters' writings reveal the critical role that preconceptions played in interpreting the limited evidence. Nebraska Man provides a valuable lesson on the importance of presumptions in interpreting evidence in the field of human origins. It also stresses the need for careful evaluation of the empirical evidence for new ideas, and the danger of going beyond what the facts warrant. PMID- 17115374 TI - The geometric side for an axiomatic theory of evolution. AB - In this paper we present a geometric model for a proposal of axiomatization of Evolution Theory. For this aim, we use suitable tools of Geometry and Topology. In particular, we define the concept of fertility factor as a main instrument for the studying of speciation. This concept, in our opinion, has an important biological meaning. PMID- 17115375 TI - The specificity enigma: from mechanics to poems. AB - Biological specificity is usually described in terms of the lock-and-key metaphor. However, this metaphor is to a certain extent misleading and does not grasp the complexity underlying biological specificity. The failure of the lock and-key metaphor makes it difficult to understand immune recognition. This is the reason why immune specificity has been described as the "Specificity Enigma." In this article, I point at three important differences between biological specificity and mechanical specificity, and suggest an alternative lens through which immune specificity can be considered. PMID- 17115376 TI - Role of canonical Wnt-signalling in joint formation. AB - The individual elements of the vertebrate skeleton are separated by three different types of joints, fibrous, cartilaginous and synovial joints. Synovial joint formation in the limbs is coupled to the formation of the prechondrogenic condensations, which precede the formation of the joint interzone. We are beginning to understand the signals involved in the formation of prechondrogenic condensations and the subsequent differentiation of cells within the condensations into chondrocytes. However, relatively little is known about the molecules and molecular pathways involved in induction of the early joint interzone and the subsequent formation of the synovial joints. Based on gain-of function studies Wnt-signalling, in particular the canonical pathway, has been implicated in the joint induction process. Here we provide genetic evidence from loss-of function analysis of embryos lacking either the central player of the canonical Wnt-pathway, beta-catenin, in the limb mesenchyme or the two ligands, Wnt9a and Wnt4, demonstrating that canonical Wnt-signalling plays an important role in suppressing the chondrogenic potential of cells in the joint thereby actively allowing joint formation. Furthermore our data show that the beta catenin activity is not essential for the induction of molecular markers expressed in the joint interzone. Thus, suggesting that canonical Wnt-signalling is not required for the induction, but for the subsequent maintenance of the fate of the joint interzone cells. PMID- 17115377 TI - Minimum proton affinity for efficient ionization with atmospheric pressure desorption/ionization on silicon mass spectrometry. AB - We performed a systematic study using a set of compounds with different proton affinities (PAs) on the ionization in atmospheric pressure desorption/ionization on silicon mass spectrometry (AP-DIOS-MS). The compounds studied included various aromatic molecules of different sizes. The PAs of these compounds were calculated using ab initio and hybrid density functional theory calculations at the B3LYP/6 31G(d) level of theory. We observed that only compounds with relatively high PAs above a threshold value of 920-950 kJ/mol were efficiently ionized as protonated molecules under AP-DIOS conditions and produced very clean mass spectra. PMID- 17115378 TI - Improved matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry of carboxymethyl cellulose. AB - A refined sample preparation procedure for matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS) was developed for the evaluation of the degree of substitution (DS) in partially depolymerised carboxymethyl cellulose (CMC). By adding ammonium sulphate to the sample mixture prior to the analysis, good quality mass spectra could be acquired. The usual time-consuming search for 'sweet-spots' at the crystalline rim of the MALDI target spot was also avoided. This quality improvement made it possible to investigate whether various positions on the target spot generated mass spectra in which the measured DS varied. The accuracy and reproducibility of the sample preparation procedure were tested by applying it on three commercial CMCs. The study shows that the DS values that were calculated from the spectra acquired from the centre region of the MALDI target spot were in better agreement with the DS provided by the supplier than were the values obtained from the large crystals at the target spot rim. This observation could be one reasonable explanation for the higher DS values reported in other publications. By applying our refined MALDI sample preparation procedure DS values that were in good agreement with the values provided by the manufacturer could be obtained. This indicates that MALDI-TOFMS of partially depolymerised CMCs can be used for an estimation of the DS as a complement to the more established methods, e.g. NMR, titrimetry, and chromatographic techniques. PMID- 17115379 TI - Langhans giant cells from M. tuberculosis-induced human granulomas cannot mediate mycobacterial uptake. AB - Tuberculosis is characterized by a tight interplay between Mycobacterium tuberculosis (M. tb) and host cells within granulomas. These cellular aggregates restrain M. tb spreading but do not kill all bacilli, which persist for years. A more detailed investigation of the interaction between M. tb and granuloma cells is needed to improve our understanding of this persistence and to explain the physiopathology of tuberculosis. In the present study, a recently developed in vitro human model of tuberculous granulomas has been used to analyse the modulation of granuloma cell differentiation by M. tb, in comparison to poorly virulent mycobacteria, which do not persist. It is reported that whilst all mycobacteria species induce granuloma formation, only M. tb triggers the differentiation of granuloma macrophages into very large multinucleated giant cells (MGCs) that are unable to mediate any bacterial uptake. This loss of function is not due to cell quiescence, as MGCs still display NADPH oxidase activity, but it correlates with decreased expression of phagocytosis receptors. This phenomenon is specific for the virulent species of M. tuberculosis complex, as poorly virulent species only induce the formation of small multinucleated cells (MCs) with conserved mycobacterial uptake ability, which never reach the MGC differentiation stage. The phenotype of MGCs thus strongly resembles mature dendritic cells with a loss of microbial uptake ability, despite conserved antigen presentation. In M. tb-induced granulomas, MGCs thus seem to be devoted to the destruction of bacilli that have been ingested in previous differentiation stages, ie in macrophages and MCs. PMID- 17115380 TI - Sumanirole versus placebo or ropinirole for the adjunctive treatment of patients with advanced Parkinson's disease. AB - The aims of this study were to assess the safety, tolerability, and efficacy of sumanirole, a highly selective D(2) dopamine receptor agonist, versus placebo in subjects with advanced Parkinson's disease (PD), and to demonstrate noninferiority of sumanirole to ropinirole. In this flexible-dose, randomized, double-blind, double-dummy, parallel-group study, 948 subjects were treated with sumanirole 1 to 48 mg/day, ropinirole 0.75 to 24 mg/day, or placebo. Treatment consisted of 13 weeks of dose escalation, 26 weeks of maintenance, and 1 week of tapering. Approximately 70% of subjects treated with either sumanirole or ropinirole completed the study. Statistical significance (P < 0.0001) was achieved when both sumanirole and ropinirole groups were compared with placebo, with mean differences of -7.7 and -8.8 on combined sum of the Unified Parkinson's Disease Rating Scale (UPDRS) part II (average on and off) and part III total scores at the end of maintenance. Noninferiority of sumanirole to ropinirole was also demonstrated, with a sumanirole minus ropinirole difference of 1.17 (90% CI: -0.56 to 2.89). Both dopamine agonists, sumanirole and ropinirole, were statistically superior compared with placebo as adjunctive therapy for patients with advanced Parkinson's disease, based on UPDRS II + III total score. Noninferiority of sumanirole to ropinirole was established, with comparable tolerability profiles. PMID- 17115381 TI - Orthostatic tremor due to thiamine deficiency. PMID- 17115382 TI - Incorporation of photodynamic therapy as an induction modality in non-small cell lung cancer. AB - BACKGROUND AND OBJECTIVES: Incorporation of photodynamic therapy (PDT) into the induction therapy regimen utilized for treatment of locally advanced primary non small cell bronchogenic carcinoma (NSCLC) is explored. STUDY DESIGN/MATERIALS AND METHODS: We present a retrospective review of 41 patients diagnosed with non metastatic NSCLC who underwent induction PDT with chemotherapy and/or radiation. RESULTS: Fifty percent of patients initially deemed unresectable, were able to undergo definitive surgical resection after trimodality induction therapy. Twenty seven percent of patients considered to require pneumonectomy were able to have a lobectomy. The pathological stage was less than the preinduction clinical stage in 14 of 22 cases; of which four patients had no residual tumor. There was no 30/90-day postoperative mortality. Mean survival was 35.9 months (lobectomy), 25.5 months (pneumonectomy) and 14.7 months (no surgery). Median survival was 78% (12 months) and 46% (36 months). The main postoperative complication following pneumonectomy was bronchopleural fistula formation. All patients who developed this complication had undergone trimodality induction therapy. Incorporation of PDT into the induction arsenal for patients with loco-regionally advanced NSCLC may be safely performed. CONCLUSIONS: PDT may define an alternative induction strategy for patients requiring pneumonectomy; further studies exploring the true efficacy of PDT as an induction modality are encouraged. PMID- 17115383 TI - An animal study of the effects on p16 and PCNA expression of repeated treatment with high-energy laser and intense pulsed light exposure. AB - BACKGROUND AND OBJECTIVE: Non-ablative skin rejuvenation treatments that involve the use of laser/light sources together with cooling devices have gained much popularity in recent years due to the lack of down time that is associated with them. One important but neglected issue is long-term safety. Does the repeated use of non-ablative skin rejuvenation lead to photoaging? Are we creating another sun-bed phenomenon? Recently, we performed an in vitro study to examine the effect of sub-lethal QS 755 nm lasers on the expression of p16INK4a on melanoma cell lines, and found that sub-lethal laser damage could increase DNA damage, which led to an increase in p16 expression. Our objective was to assess the cutaneous effect of repeated exposure to high-energy lasers and intense pulsed light sources on male Institute of Cancer Research (ICR) mice. STUDY DESIGN/MATERIALS AND METHODS: Twenty-eight male ICR mice were divided into four groups. Other than the control group, all groups received either laser (585 nm pulsed dye laser or 1,320 nm Nd:YAG laser) or intense pulsed light (IPL) treatment. All four groups were anesthetized with a mixture of Hypnorm/Dormicum before treatment. The animals were irradiated twice a week for 6 months. Signs of toxicity such as mortality and weight loss were checked once a week. Skin tumor formation was evidenced by lesions of greater than 1 mm in diameter that persisted for 2 weeks. At the end of the 6 months, the expression of proliferating cell nuclear antigen (PCNA) and p16 in the mouse skin was determined by immunohistochemical staining and immunoblotting using specific monoclonal antibodies for mouse PCNA and p16. The results were expressed as mean +/- standard error of the mean (SEM). Statistical difference was assessed by multiple ANOVA. A P-value of <0.05 was considered to be significant. RESULTS: At the end of the 6 months, none of the animals had developed any signs of toxicity such as mortality or weight lost. There was no evidence of tumor formation. There were significant elevations of p16 and PCNA in all treated groups as compared to the control group (ANOVA P < 0.05). This particularly applied to the group that was treated with the 1,320 nm Nd:YAG laser. CONCLUSION: The repeated use of high energy laser and intense pulsed light source did not cause any toxicity in mice. The changes in p16 and PCNA imply that further studies are necessary to consider the implications of repeated exposure to longer wavelength radiation in human skin. PMID- 17115384 TI - Ex vivo histological characterization of a novel ablative fractional resurfacing device. AB - BACKGROUND AND OBJECTIVES: We introduce a novel CO(2) laser device that utilizes ablative fractional resurfacing for deep dermal tissue removal and characterize the resultant thermal effects in skin. STUDY DESIGN/MATERIALS AND METHODS: A prototype 30 W, 10.6 microm CO(2) laser was focused to a 1/e(2) spot size of 120 microm and pulse duration up to 0.7 milliseconds to achieve a microarray pattern in ex vivo human skin. Lesion depth and width were assessed histologically using either hematoxylin & eosin (H&E) or lactate dehydrogenase (LDH) stain. Pulse energies were varied to determine their effect on lesion dimensions. RESULTS: Microarrays of ablative and thermal injury were created in fresh ex vivo human skin irradiated with the prototype CO(2) laser device. Zones of tissue ablation were surrounded by areas of tissue coagulation spanning the epidermis and part of the dermis. A thin condensed lining on the interior wall of the lesion cavity was observed consistent with eschar formation. At 23.3 mJ, the lesion width was approximately 350 microm and depth 1 mm. In this configuration, the cavities were spaced approximately 500 microm apart and interlesional epidermis and dermis demonstrated viable tissue by LDH staining. CONCLUSION: A novel prototype ablative CO(2) laser device operating in a fractional mode was developed and its resultant thermal effects in human abdominal tissue were characterized. We discovered that controlled microarray patterns could be deposited in skin with variable depths of dermal tissue ablation depending on the treatment pulse energy. This is the first report to characterize the successful use of ablative fractional resurfacing as a potential approach to dermatological treatment. PMID- 17115385 TI - Spatial refractive index measurement of porcine artery using differential phase optical coherence microscopy. AB - BACKGROUND AND OBJECTIVES: We describe a methodology to record spatial variation of refractive index of porcine renal artery using differential phase optical coherence microscopy (DP-OCM). STUDY DESIGN/MATERIALS AND METHODS: The DP-OCM provides quantitative measurement of thin specimen phase retardation and refractive index by measuring optical path-length changes on the order of a few nanometers and with a lateral resolution of 3 microm. The DP-OCM instrumentation is an all-fiber, dual-channel Michelson interferometer constructed using a polarization maintaining (PM) fiber. RESULTS: Two-dimensional en face dual channel phase images are taken over a 150 x 200 microm region on a microscopic slide, and the images are reconstructed by plotting a two-dimensional refractive index map as the OCM beam is moved across the sample. CONCLUSIONS: Because the DP OCM can record transient changes in the optical path-length, the system may be used to record quantitative optical path-length alterations of tissue in response to various stimuli. A fiber-based DP-OCM may have the potential to substantially improve in vivo imaging of individual cells for a variety of clinical diagnostics, and monitoring applications. PMID- 17115386 TI - Autopsy-proven Huntington's disease with 29 trinucleotide repeats. AB - Huntington's disease (HD) is a neurodegenerative disorder associated with expansion of CAG trinucleotide repeats in the huntingtin gene. A minimum of 36 CAG repeats is usually reported in patients with clinical features of HD; 30 to 35 repeats represent an intermediate range. Here we report a 65-year-old male with autopsy-proven HD and 29 CAG repeats. PMID- 17115387 TI - Movement Disorder Society-sponsored revision of the Unified Parkinson's Disease Rating Scale (MDS-UPDRS): Process, format, and clinimetric testing plan. AB - This article presents the revision process, major innovations, and clinimetric testing program for the Movement Disorder Society (MDS)-sponsored revision of the Unified Parkinson's Disease Rating Scale (UPDRS), known as the MDS-UPDRS. The UPDRS is the most widely used scale for the clinical study of Parkinson's disease (PD). The MDS previously organized a critique of the UPDRS, which cited many strengths, but recommended revision of the scale to accommodate new advances and to resolve problematic areas. An MDS-UPDRS committee prepared the revision using the recommendations of the published critique of the scale. Subcommittees developed new material that was reviewed by the entire committee. A 1-day face-to face committee meeting was organized to resolve areas of debate and to arrive at a working draft ready for clinimetric testing. The MDS-UPDRS retains the UPDRS structure of four parts with a total summed score, but the parts have been modified to provide a section that integrates nonmotor elements of PD: I, Nonmotor Experiences of Daily Living; II, Motor Experiences of Daily Living; III, Motor Examination; and IV, Motor Complications. All items have five response options with uniform anchors of 0 = normal, 1 = slight, 2 = mild, 3 = moderate, and 4 = severe. Several questions in Part I and all of Part II are written as a patient/caregiver questionnaire, so that the total rater time should remain approximately 30 minutes. Detailed instructions for testing and data acquisition accompany the MDS-UPDRS in order to increase uniform usage. Multiple language editions are planned. A three-part clinimetric program will provide testing of reliability, validity, and responsiveness to interventions. Although the MDS UPDRS will not be published until it has successfully passed clinimetric testing, explanation of the process, key changes, and clinimetric programs allow clinicians and researchers to understand and participate in the revision process. PMID- 17115388 TI - Acute effects of immediate and controlled-release levodopa on mood in Parkinson's disease: A double-blind study. AB - Mood fluctuations related to levodopa (LD) dosing are well-known psychiatric complications of Parkinson's disease (PD). No formal studies explored how affective response to LD relates to the type of motor response to oral LD (stable or wearing-off) and to different pharmacokinetic profiles of oral LD. We used an intrasubject randomized double-blind crossover design to study 14 patients (7 stable, 7 wearing-off) who were monitored for motor status, mood, anxiety, and plasma LD levels 1 hour before and 6 hours after an oral dose of immediate release (IR) and controlled-release LD formulations. Analysis of the dose response curves showed a significant interaction between the type of motor response and the type of LD. Only the wearing-off patients had a significant mood elevation, and this effect was only significant following challenge with IR LD. Motor status strongly correlated with LD plasma levels and anxiety but not with mood ratings. Mood changes in PD patients are related to the patient's type of motor response to oral LD and also to the kinetic profile of the LD formulation used for dopaminergic replacement. PMID- 17115389 TI - Angiogenesis inhibition with bevacizumab and the surgical management of colorectal cancer. AB - BACKGROUND: Bevacizumab is an angiogenesis inhibitor and a new therapy for the treatment of colorectal cancer. It is a humanized monoclonal antibody that targets vascular endothelial growth factor. METHODS: This review is based on a literature search of Medline, Pubmed, ISI web of knowledge and other published work for original articles, reviews and abstracts relevant to the surgical management of colorectal cancer with bevacizumab. RESULTS AND CONCLUSION: Combined with current chemotherapy regimens, bevacizumab offers a significant survival advantage, making it likely to see widespread use. Despite being generally well tolerated, serious toxicities, including wound complications and gastrointestinal perforation, have been reported that affect surgical management. Consideration should be given to the timing of surgical and adjuvant intervention when using this drug. PMID- 17115390 TI - Randomized clinical trial of 0.2 per cent glyceryl trinitrate ointment for wound healing and pain reduction after open diathermy haemorrhoidectomy. AB - BACKGROUND: Open haemorrhoidectomy is associated with considerable postoperative pain and discomfort. This study assessed whether glyceryl trinitrate (GTN) ointment promotes wound healing and reduces pain after open haemorrhoidectomy. METHODS: A randomized prospective double-blind placebo-controlled trial was conducted. Patients were randomized to either 0.2 per cent GTN ointment or placebo ointment (petroleum jelly). Patients were asked to fill in a pain diary. Complete healing was defined as complete epithelialization. RESULTS: There were 40 patients in the GTN group and 42 in the placebo group. There were no statistically significant differences in sex, weight, type of haemorrhoid, type of surgery (emergency or elective), number of haemorrhoids excised, duration of surgery, hospital stay and complication rate between the groups. Pain scores and analgesic use were not significantly different. By week 3, however, 17 patients in the GTN group had completely epithelialized wounds compared with eight patients in the placebo group (P = 0.021). Only one patient who received GTN experienced headache requiring discontinuation of the ointment. CONCLUSION: TGN 0.2 per cent ointment improved wound healing rates, but did not reduce pain in this study. PMID- 17115393 TI - Laryngeal dystonia as a presenting symptom of young-onset Parkinson's disease. PMID- 17115391 TI - Screening for Lrrk2 G2019S and clinical comparison of Tunisian and North American Caucasian Parkinson's disease families. AB - Mutations in the leucine-rich repeat kinase-2 gene (LRRK2) are responsible for some forms of familial as well as sporadic Parkinson's disease (PD). The purpose of this study was to examine the frequency of a single pathogenic mutation (6055G > A) in the kinase domain of this gene in United States and Tunisian familial PD and to compare clinical characteristics between patients with and without the mutation. Standardized case report forms were used for clinical and demographic data collection. We investigated the frequency of the most common substitution of LRRK2 (G2019S, 6055G>A) and its impact on epidemiological and phenotypic features. The frequency of mutations in Tunisian families was 42% (38/91) and in U.S. families 2.6% (1/39), with the unique opportunity to compare homozygous (n = 23) and heterozygous (n = 109) Tunisian carriers of G2019S substitutions. Individuals with G2019S substitutions had an older age at onset but few other differences compared with families negative for the substitution. Patients with LRRK2 mutations had typical clinical features of PD. Comparisons between individuals with heterozygous and homozygous LRRK2 mutations suggested that gene dosage was not correlated with phenotypic differences; however, the estimated penetrance was greater in homozygotes across all age groups. PMID- 17115394 TI - Evidence-based guidelines for using the Short Form 36 in cervical dystonia. AB - We aimed to provide evidence-based guidelines for using the Short Form 36 (SF-36) as an outcome measure in cervical dystonia (CD). To do this, we tested the hypothesized relationships between items, scales, and summary measures of the SF 36 using psychometric analyses in data from a postal survey of 235 people with CD. Although the majority of subscales performed adequately, the Role Physical and Role Emotional subscales had substantial floor and/or ceiling effects. Evidence did not support computing SF-36 Physical and Mental Component Summary scores. We propose guidelines that include the recommendation that these subscale and summary scores should be reported with caution. PMID- 17115395 TI - A tale of three papers. PMID- 17115396 TI - Valproate-induced Parkinsonism in epilepsy patients. AB - We systematically examined 226 epilepsy patients in a tertiary-referral center and found 6 (5.04%) to have valproate-induced Parkinsonism. There was a significantly higher prevalence of patients with Parkinsonism in the group of patients treated with valproate compared to those who were on other antiepileptic drugs (6 [5.04%] of 119 vs. 0 [0%] of 107; chi2 = 5.54; P = 0.025). These six patients had been on valproate for more than 3 years (mean, 75.67 +/- 25.32 months) at an average dose of 750 +/- 273.86 mg/day. The valproate doses were decreased or discontinued with supplementation from another antiepileptic medication. The mean UPDRS motor score significantly improved from 10.67 +/- 5.1 to 4.75 +/- 2.75 (P < 0.05). There was no relapse of seizures. Clinicians working in tertiary-referral centers should have a high index of suspicion for valproate induced Parkinsonism. Early recognition and switching into another antiepileptic medication may help reduce unnecessary suffering in these patients. PMID- 17115397 TI - Randomized clinical trial of bowel preparation with a single phosphate enema or polyethylene glycol before elective colorectal surgery (Br J Surg 2006; 93; 427 433). PMID- 17115398 TI - Randomized clinical trial assessing the effect of Doppler-optimized fluid management on outcome after elective colorectal resection (Br J Surg) 2006; 93; 1069-1076. PMID- 17115399 TI - Randomized clinical trial of the effect of quilting latissimus dorsi flap donor site on seroma formation (Br J Surg) 2006; 93; 825-830. PMID- 17115400 TI - Re: clinical feature profile of spinocerebellar ataxia type 1-8 predicts genetically defined subtypes. PMID- 17115402 TI - Neoadjuvant chemotherapy and resection of advanced synchronous liver metastases before treatment of the colorectal primary (Br J Surg) 2006; 93; 872-878. PMID- 17115404 TI - Enhanced degradation of atrazine under field conditions correlates with a loss of weed control in the glasshouse. AB - Enhanced degradation of atrazine has been reported in the literature, indicating the potential for reduced residual weed control with this herbicide. Experiments were conducted to determine the field dissipation of atrazine in three cropping systems: continuous Zea mays L. (CC) receiving atrazine applications each year, Gossypium hirsutum L.-Z. mays rotation (CCR) receiving applications of atrazine once every 2 years and a no atrazine history soil (NAH). Subsequent laboratory and greenhouse experiments were conducted with soil collected from these cropping systems to determine atrazine degradation, mineralization and residual weed control. Field dissipation of atrazine followed first-order kinetics, and calculated half-life values for atrazine combined over 2003 and 2005 increased in the order of CC (9 d) = CCR (10 d) < NAH (17 d). Greenhouse studies confirmed that the persistence of atrazine was approximately twofold greater in NAH soil than in CC or CCR soil. Biometer flask mineralization studies suggested that enhanced degradation of atrazine was due to rapid catabolism of the s-triazine ring. Glasshouse efficacy studies revealed a loss of residual weed control in CC and CCR soil compared with NAH soil. These data indicate that, under typical Mississippi Delta field conditions and agronomic practices, the persistence of atrazine may be reduced by at least 50% if the herbicide is applied more than once every 24 months. Glasshouse studies suggest that under these conditions a loss of residual weed control is possible. PMID- 17115406 TI - Adjuvant therapy for breast cancer. PMID- 17115408 TI - Peutz-Jeghers syndrome and screening for pancreatic cancer. AB - BACKGROUND: Cancer risk, including pancreatic, is high in those with Peutz Jeghers syndrome (PJS). It has been suggested that such patients should undergo screening for pancreatic cancer. METHODS: The risk of pancreatic cancer in PJS, pancreatic screening and potential screening strategies were reviewed. Cost effectiveness was assessed according to American Gastroenterology Association guidelines and a risk stratification model proposed by the European Registry of Hereditary Pancreatitis and Familial Pancreatic Cancer. RESULTS: The risk of pancreatic cancer is increased in PJS but screening would cost over US 35,000 dollars per life saved. Risk stratification reduces cost by 100,000 dollars and costs fall to 50,000 dollars per life saved if deaths from other forms of cancer are avoided. CONCLUSION: Screening should be performed only on a research basis to evaluate the benefit and cost-effectiveness in high-risk groups. PMID- 17115409 TI - Vascular permeability induced by VEGF family members in vivo: role of endogenous PAF and NO synthesis. AB - We previously reported that vascular endothelial growth factor (VEGF) increases vascular permeability through the synthesis of endothelial platelet-activating factor (PAF), while others reported the contribution of nitric oxide (NO). Herein, we addressed the contribution of VEGF receptors and the role played by PAF and NO in VEGF-induced plasma protein extravasation. Using a modified Miles assay, intradermal injection in mice ears of VEGF-A(165), VEGF-A(121), and VEGF-C (1 microM) which activate VEGFR-2 (Flk-1) receptor increased vascular permeability, whereas a treatment with VEGFR-1 (Flt-1) analogs; PlGF and VEGF-B (1 microM) had no such effect. Pretreatment of mice with PAF receptor antagonist (LAU8080) or endothelial nitric oxide synthase (eNOS) inhibitor (L-NAME) abrogated protein extravasation mediated by VEGF-A(165). As opposed to PAF (0.01 1 microM), treatment with acetylcholine (ACh; up to 100 microM; inducer of NO synthesis) or sodium nitroprusside (SNP; up to 1 microM; NO donor) did not induce protein leakage. Simultaneous pretreatment of mice with eNOS and protein kinase A (PKA) inhibitors restored VEGF-A(165) vascular hyperpermeability suggesting that endogenous NO synthesis leads to PKA inhibition, which support maintenance of vascular integrity. Our data demonstrate that VEGF analogs increase vascular permeability through VEGFR-2 activation, and that both endogenous PAF and NO synthesis contribute to VEGF-A(165)-mediated vascular permeability. However, PAF but not NO directly increases vascular permeability per se, thereby, suggesting that PAF is a direct inflammatory mediator, whereas NO serves as a cofactor in VEGF-A(165) proinflammatory activities. PMID- 17115410 TI - Mouse embryonic fibroblast cells from transgenic mice overexpressing tNOX exhibit an altered growth and drug response phenotype. AB - Mouse embryonic fibroblast (MEF) cells prepared from transgenic mice overexpressing a cancer-specific and growth-related cell surface NADH oxidase with protein disulfide-thiol interchange activity grew at rates approximately twice those of wild-type embryonic fibroblast cells. Growth of transgenic MEF cells overexpressing tNOX was inhibited by low concentrations of the green tea catechin (-)-epigallocatechin-3-gallate (EGCg) or the synthetic isoflavene phenoxodiol. Both are putative tNOX-targeted inhibitors with anti-cancer activity. With both EGCg and phenoxodiol, growth inhibition was followed after about 48 h by apoptosis. Growth of wild-type mouse fibroblast cells from the same strain was unaffected by EGCg and phenoxodiol and neither compound induced apoptosis even at concentrations 100-1,000-fold higher than those that resulted in apoptotic death in the transgenic MEF cells. The findings validate earlier reports of evidence for tNOX presence as contributing to unregulated growth of cancer cells as well as the previous identification of the tNOX protein as the molecular target for the anti-cancer activities attributed to both EGCg and phenoxodiol. The expression of tNOX emerges as both necessary and sufficient to account for the cancer cell-specific growth inhibitions by both EGCg and phenoxodiol. PMID- 17115411 TI - Approach to systematic analysis of serine/threonine phosphoproteome using Beta elimination and subsequent side effects: intramolecular linkage and/or racemisation. AB - Complete analysis of the phosphorylation of serine and threonine residues directly from biological extracts is still at an early stage and will remain a challenging goal for many years. Analysis of phosphorylated proteins and identification of the phosphorylated sites in a crude biological extract is a major topic in proteomics, since phosphorylation plays a dominant role in post translational protein modification. Beta elimination of the serine/threonine bound phosphate by alkali action generates (methyl)dehydroalanine. The reactivity of this group susceptible of nucleophilic attacks might be used as a tool for phosphoproteome analysis. Most of the known serine/threonine kinases recognize motifs in protein targets that are rich in lysine(s) and/or arginine(s). The (methyl)dehydroalanine resulting from beta elimination of the serine/threonine bound phosphate by alkali action is likely to react with the amino groups of these neighboring amino acids. Furthermore, the addition reaction of dehydroalanine-peptides with a nucleophilic group more likely generates diastereoisomers derivatives. The internal cyclic bonds and/or the stereoisomer peptide derivatives thus generated confer resistance to trypsin cleavage and/or constitute stop signals for exopeptidases such as carboxypeptidase. This might form the basis of a method to facilitate the systematic identification of phosphorylated peptides. PMID- 17115412 TI - Hyaline cartilage formation and enchondral ossification modeled with KUM5 and OP9 chondroblasts. AB - What is it that defines a bone marrow-derived chondrocyte? We attempted to identify marrow-derived cells with chondrogenic nature and immortality without transformation, defining "immortality" simply as indefinite cell division. KUM5 mesenchymal cells, a marrow stromal cell line, generated hyaline cartilage in vivo and exhibited enchondral ossification at a later stage after implantation. Selection of KUM5 chondroblasts based on the activity of the chondrocyte-specific cis-regulatory element of the collagen alpha2(XI) gene resulted in enhancement of their chondrogenic nature. Gene chip analysis revealed that OP9 cells, another marrow stromal cell line, derived from macrophage colony-stimulating factor deficient osteopetrotic mice and also known to be niche-constituting cells for hematopoietic stem cells expressed chondrocyte-specific or -associated genes such as type II collagen alpha1, Sox9, and cartilage oligomeric matrix protein at an extremely high level, as did KUM5 cells. After cultured OP9 micromasses exposed to TGF-beta3 and BMP2 were implanted in mice, they produced abundant metachromatic matrix with the toluidine blue stain and formed type II collagen positive hyaline cartilage within 2 weeks in vivo. Hierarchical clustering and principal component analysis based on microarray data of the expression of cell surface markers and cell-type-specific genes resulted in grouping of KUM5 and OP9 cells into the same subcategory of "chondroblast," that is, a distinct cell type group. We here show that these two cell lines exhibit the unique characteristics of hyaline cartilage formation and enchondral ossification in vitro and in vivo. PMID- 17115413 TI - VDR-mediated gene expression patterns in resting human coronary artery smooth muscle cells. AB - Vitamin D analogs such as paricalcitol and calcitriol that activate the vitamin D receptor (VDR) provide survival benefit for Stage 5 chronic kidney disease (CKD) patients, possibly associated with a decrease in cardiovascular (CV)-related incidents. Phenotypic changes of smooth muscle cells play an important role in CV disease. The role of vitamin D analogs in modulating gene expression in smooth muscle cells is still not well understood. In this study, DNA microarray analysis of approximately 22,000 different human genes was used to characterize the VDR mediated gene expression profile in human coronary artery smooth muscle cells (CASMC) at rest. Cells in serum free medium were treated with 0.1 microM calcitriol (1alpha,25-dihydroxyvitamin D(3)) or paricalcitol (19-nor-1alpha,25 (OH)(2)D(2)) for 30 h. A total of 181 target genes were identified, with 103 genes upregulated and 78 downregulated (>two fold changes in either drug treatment group with P < 0.01). No significant difference was observed between calcitriol and paricalcitol. Target genes fell into various categories with the top five in cellular process, cell communication, signal transduction, development, and morphogenesis. Twenty-two selected genes linked to the CV system were also impacted. Real-time RT-PCR and/or Western blotting analysis were employed to confirm the expression patterns of selected genes such as 25 hydroxyvitamin D-24-hydroxylase, Wilms' tumor gene 1, transforming growth factorbeta3, plasminogen activator inhibitor-1, thrombospondin-1 (THBS1), and thrombomodulin (TM). This study provides insight into understanding the role of VDR in regulating gene expression in resting smooth muscle cells. PMID- 17115414 TI - What is the function of centrioles? AB - The function of centrioles has been controversial and remains incompletely resolved. This is because centrioles, in and of themselves, do not directly perform any physiological activity. Instead, their role is only to act as a jig or breadboard onto which other functional structures can be built. Centrioles are primarily involved in forming two structures-centrosomes and cilia. Centrioles bias the position of spindle pole formation, but because spindle poles can self organize, the function of the centriole in mitosis is not obligatory. Consequently, lack of centrioles does not generally prevent mitosis, although recent experiments suggest acentriolar spindles have reduced fidelity of chromosome segregation. In contrast, centrioles are absolutely required for the assembly of cilia, including primary cilia that act as cellular antennae. Consistent with this requirement, it is now becoming clear that many ciliary diseases, including nephronophthisis, Bardet-Biedl syndrome, Meckel Syndrome, and Oral-Facial-Digital syndrome, are caused by defects in centriole-associated proteins. PMID- 17115415 TI - Palladin regulates cell and extracellular matrix interaction through maintaining normal actin cytoskeleton architecture and stabilizing beta1-integrin. AB - Cell and extracellular matrix (ECM) interaction plays an important role in development and normal cellular function. Cell adhesion and cell spreading on ECM are two basic cellular behaviors related to cell-ECM interaction. Here we show that palladin, a novel actin cytoskeleton-associated protein, is actively involved in the regulation of cell-ECM interaction. It was found that palladin deficient mouse embryonic fibroblasts (MEFs) display decreased cell adhesion and compromised cell spreading on various ECMs. Disorganized actin cytoskeleton architecture characterized by faint stress fibers, less lamellipodia and focal adhesions can account for the weakened cell-ECM interaction in palladin(-/-) MEFs. Furthermore, decreased polymerized filament actin and increased globular actin can be observed in palladin(-/-) MEFs, strongly suggesting that palladin is essential for the formation or stabilization of polymerized filament actin. Elevated phospho-cofilin level and proper responses in cofilin phosphorylation to either Rho signal agonist or antagonist in palladin(-/-) MEFs indicate that disrupted stress fibers in palladin(-/-) MEFs is not associated with cofilin phosphorylation. More interestingly, the protein level of ECM receptor beta1 integrin is dramatically decreased in MEFs lacking palladin. Down-regulation of beta1-integrin protein can be restored by proteasome inhibitor MG-132 treatment. All these data implicate that palladin is essential for cell-ECM interaction through maintaining normal actin cytoskeleton architecture and stabilizing beta1 integrin protein. PMID- 17115416 TI - PACAP/PAC1 autocrine system promotes proliferation and astrogenesis in neural progenitor cells. AB - The Pituitary adenylate cyclase-activating peptide (PACAP) ligand/type 1 receptor (PAC1) system regulates neurogenesis and gliogenesis. It has been well established that the PACAP/PAC1 system induces differentiation of neural progenitor cells (NPCs) through the Gs-mediated cAMP-dependent signaling pathway. However, it is unknown whether this ligand/receptor system has a function in proliferation of NPCs. In this study, we identified that PACAP and PAC1 were highly expressed and co-localized in NPCs of mouse cortex at embryonic day 14.5 (E14.5) and found that the PACAP/PAC1 system potentiated growth factor-induced proliferation of mouse cortical NPCs at E14.5 via Gq-, but not Gs-, mediated PLC/IP3-dependent signaling pathway in an autocrine manner. Moreover, PAC1 activation induced elongation of cellular processes and a stellate morphology in astrocytes that had the bromodeoxyuridine (BrdU)-incorporating ability of NPCs. Consistent with this notion, we determined that the most BrdU positive NPCs differentiated to astrocytes through PAC1 signaling. These results suggest that the PACAP/PAC1 system may play a dual role in neural/glial progenitor cells not only differentiation but also proliferation in the cortical astrocyte lineage via Ca2+-dependent signaling pathways through PAC1. PMID- 17115417 TI - IL-2 gene C/T polymorphism is associated with prostate cancer. AB - Cytokines are reported to be associated with the formation of prostate cancer. Our aim was to investigate whether C/T polymorphisms of the interleukin-2 (IL-2) gene and IL-2 receptor beta (IL-2RB) gene are associated with prostate cancer. We compared the frequency of the polymorphisms of the IL-2 gene and the IL-2RB gene between 96 patients with prostate cancer and 105 healthy male volunteers from the same area (age >60 years). They were followed for at least 5 years. There was a significant difference in distribution of the genotype of the IL-2 gene polymorphism between the prostate cancer group and the control group (P = 0.017). The distribution of the TT homozygote of the IL-2 gene was significantly higher in the cancer group (32.3%) than in the control group (16.2%). However, no significant statistical difference was found between the polymorphism of the IL-2 gene and prostate cancer in survival analysis during a 5-year follow up period (log rank test; P = 0.19). There was no significant difference in the distribution of the genotype of the IL-2RB gene polymorphism between controls and cancer patients (P = 0.388). This study suggests that the IL-2 gene may be associated with susceptibility to prostate cancer in the Taiwan population. PMID- 17115418 TI - Establishment of a flow cytometric assay for determination of human platelet glycoprotein VI based on a mouse polyclonal antibody. AB - Glycoprotein VI (GPVI) is the major signaling receptor for collagen on platelets. Recent studies suggest that the surface density of GPVI is related to the activation of platelets by collagen. To measure the level of GPVI on platelets, a mouse polyclonal antibody BJ010 was prepared using an amplified fragment of extracellular domain in GPVI. The specific reactivity of BJ010 was identified by anti-GPVI specific monoclonal antibody 11a12 using immunoprecipitation, sandwich enzyme-linked immunosorbent assay (ELISA) and flow cytometry. The antibody BJ010 recognized both native and denatured human GPVI so that it was used to set up a flow cytometric assay to detect the level of GPVI in normal subjects. The relative level of GPVI on platelets was detected in 101 healthydonors. The median geometric mean fluorescence intensity (GMFI) of platelet GPVI level was 57.7. The variation between minimum and maximum values of platelet GPVI and integrin alpha2beta1 were found to be 3.5- and 4.1-fold, respectively, in the normal subjects. There was a week correlation between the amount of GPVI and integrin alpha2beta1 on platelet surfaces. For the method, the intraassay and interassay coefficient of variation was 6.3% and 8.8%, respectively. The flow cytometric assay described here provides a simple, reliable, reproducible, and readily available means of quantitation of collagen receptor GPVI density on the platelet surface in a larger number of blood samples. PMID- 17115419 TI - Proinflammatory cytokine gene polymorphisms among Hashimoto's thyroiditis patients. AB - Proinflammatory cytokines are involved in the pathogenesis of Hashimoto's thyroiditis (HT). To test whether certain specific proinflammatory cytokine gene polymorphisms could be genetic markers for an individual's susceptibility to HT, we investigated single-site polymorphisms of certain proinflammatory cytokine genes of interest for 107 HT sufferers and 163 controls, subsequent to preparing the necessary experimental genomic DNA from peripheral blood, using a polymerase chain reaction (PCR)-based restriction analysis. The polymorphisms we detected were as follows: 1) C/T and E1/E2 polymorphisms for the interleukin (IL)-1beta gene at promoter (-511) and exon 5, respectively; 2) a variable number of tandem repeats (VNTRs) for the IL-1 receptor antagonist (IL-1Ra) gene at intron 2; 3) a C/G polymorphism for the IL-6 gene at promoter (-572); and 4) an A/G polymorphism for the tumor necrosis factor (TNF)-alpha gene at promoter (-308). The data demonstrated an increased ratio of CG genotype and decreased ratios of CC and GG genotypes (chi-squared test; P = 0.025) for the IL-6 gene promoter for HT patients when compared with normal controls. The odds ratio (OR) for the CG genotype, as compared to the GG genotype, for HT patients was shown to be 4.065 (95% confidence interval (CI): 1.268-13.032). Comparison of the genotype analysis for the remaining gene polymorphisms and the allelic analysis for all of the screened gene polymorphisms, however, all revealed no statistically significant difference between the two study groups as regards frequency of genotype. In conclusion, we suggest that an IL-6 gene promoter (-572) C/G polymorphism could represent a potential "candidate" genetic marker to predict an individual's susceptibility to HT. PMID- 17115420 TI - Comparison of BNP and NT-proBNP assays in the approach to the emergency diagnosis of acute dyspnea. AB - N-terminal pro-brain natriuretic peptide (NT-proBNP) and BNP measurement could have a significant role in differentiating dyspnea between cardiac or pulmonary origin in the emergency room. The development of new and different commercial assays for these B-type natriuretic peptides offers the possibility of improving and simplifying their measurements but this could be defaulted due to the differences in methodology and the lack of assay standardization. We compared four available methods of measuring NT-proBNP and BNP and evaluated their usefulness in diagnosing the causes of breathlessness in the emergency room. The correlation of BNP with different assays was strong with r>0.98 (P<0.0001). Comparison studies between NT-proBNP and BNP procedures were in good agreement with r>0.87. The area under the receiver-operating characteristic curve (ROC) for BNP or NT-proBNP for detecting any cardiac dysfunction was higher than 0.96 (95% CI). A BNP value of 116 pg/mL measurement with the Access BNP assay (Beckman Coulter Inc., Fullerton, CA), a BNP value of 79 pg/mL with Advia Centaur BNP assay (Bayer Diagnostics, Tarrytown, NY), and an NT-proBNP level of 817 pg/mL in Elecsys NT-proBNP assay (Roche Diagnostic, Mannheim, Germany), showed both high sensitivity (>92%) and high specificity (>93%). We have found that NT-proBNP and BNP present similar diagnostic accuracies for the differential diagnosis of dyspnea. PMID- 17115421 TI - Switching from HPLC/UV to MEIA for whole blood sirolimus quantitation: comparison of methods. AB - Sirolimus is a immunosuppressive agent for renal transplant recipients. Monitoring of whole blood sirolimus concentration is necessary in order to improve clinical outcomes. An increasing number of clinical laboratories (4-14% during 2005) are using microparticle enzyme immunoassay (MEIA) for sirolimus quantitation but previous reports indicated a high variability, with a mean difference of 17% for MEIA method vs. high-performance liquid chromatography/ultraviolet (HPLC/UV). This study was aimed at comparing the reliability of MEIA with the HPLC/UV method. Blood samples from transplant patients were processed using both HPLC/UV and MEIA assays. Comparison and Bland Altman plots, as well as regression analysis and paired t-test were used to compare results of the assays. Concentrations were stratified into three groups and used to investigate whether any observed difference between methods could be influenced by sirolimus concentration. Regression analysis yielded a coefficient of correlation R of 0.9756, the line of best fit being y=0.9832x+0.1976. The statistical analysis showed no difference between the two sets of experimental data. The average percentage difference between the two methods was found to be 0.2+/-19.2%. On the basis of our present results, the tested MEIA assay is able to quantify sirolimus concentration with a clinically acceptable imprecision, similar to that of HPLC/UV method. PMID- 17115422 TI - Lack of association of interleukin-6 and interleukin-8 gene polymorphisms in Chinese patients with systemic lupus erythematosus. AB - The purpose of this study was to determine whether interleukin (IL)-6 and IL-8 gene polymorphisms were markers of susceptibility to or severity of systemic lupus erythematosus (SLE) in Chinese patients. The study included 150 Chinese patients with SLE. A total of 130 unrelated healthy individuals living in central Taiwan served as control subjects. Polymorphisms of the IL-6 and IL-8 gene were typed from genomic DNA. The genotypes, allelic frequencies, and carriage rates were compared between SLE patients and control subjects. The relationship between allelic frequencies and clinical manifestations of 135 SLE patients was evaluated. There were no statistically significant differences in IL-6 and IL-8 gene polymorphisms between the SLE and control groups (chi-squared test, P=0.53, chi(2)=1.27 and P=0.44, chi(2)=1.62, respectively). In addition, there was no significant association between the two groups in allelic frequency of IL-6 and IL-8 (P=0.89 and P=0.26, respectively). We also did not detect any association between the IL-6 and IL-8 genotype and clinical or laboratory profiles in SLE patients. The results suggest that the IL-6 and IL-8 gene polymorphisms are not related to SLE. PMID- 17115423 TI - Evaluation of the Treponema pallidum particle agglutination technique (TP.PA) in the diagnosis of neurosyphilis. AB - The Treponema pallidum particle agglutination technique (TP.PA) was evaluated, in comparison with the Venereal Disease Research Laboratory (VDRL) test, microhemagglutination assay for Treponema pallidum antibodies (MHA-TP), and fluorescent treponemal antibody-ABS (FTA-Abs) test for the diagnosis of neurosyphilis. We have studied 198 cerebrospinal fluid (CSF) samples from patients with syphilis, including neurosyphilis, treated syphilis, and with other neurological manifestations than neurosyphilis. All tests were nonreactive in these last group of patients. In the neurosyphilis patients, sensitivity of the TP.PA was 100%. The performance of this test in CSF from patients with primary syphilis was as good as that of the other tests. In secondary and latent syphilis, the TP.PA results (27 reactive samples/73) were similar to those of the MHA-TP (25 reactive samples/73). In the individuals treated for syphilis, the TP.PA, FTA-Abs, and MHA-TP tests were found to be reactive in eight, six, and eight samples, respectively. In conclusion, it seems that the TP.PA can be used in CSF to diagnose neurosyphilis, although as for other serological tests, interpretation of results should be done in conjunction with other neurosyphilis parameters. PMID- 17115424 TI - Darbepoetin alpha for the treatment of anemia in patients with myelodysplastic syndromes. AB - BACKGROUND: Anemia occurs as a comorbidity in from 80% to 85% of patients with myelodysplastic syndromes (MDS): It causes fatigue, increases transfusion needs, and reduces quality of life. Darbepoetin alpha (DA) is an erythropoiesis stimulating protein (ESP) that is more highly glycosylated and has a longer half life relative to recombinant human erythropoietin (rHuEPO), thus, allowing less frequent administration, increased convenience, and better compliance. METHODS: This retrospective analysis included 81 patients with MDS who were enrolled at 9 Spanish centers and who received once-weekly, subcutaneous DA (75-300 microg) for 16 weeks. RESULTS: Fifty-five percent of all patients (38 of 69 evaluable patients) achieved responses; 30.4% of were major responses, and 24.6% were minor responses; 64.7% of rHuEPO-naive patients and 45.7% rHuEPO-treated patients responded; and 43.2% had received previous rHuEPO. Most responses (65.8%) occurred at or before Week 8. The median age at diagnosis was 70 years (range, 38 87 years), the median age at the initiation of DA treatment was 75 years (range, 39-91 years), and 56.8% of patients were women. The median time from last ESP dose to DA initiation was 16.8 weeks (range, 0.0-159.0 weeks; <1 week in 53.1% of patients). According to the French-American-British classification system (n = 81 patients), 39.5% had refractory anemia (RA), 46.9% had RA with ringed sideroblasts, 9.9% had RA with excess blasts (RAEB), 1.2% had RAEB in transformation, and 2.5% had chronic myelomonocytic leukemia. According to the International Prognostic Scoring System (n = 47 patients), 55.3% of patients were in the low-risk group, and 36.2% of patients were in the intermediate-1-risk group. The median baseline hemoglobin level was 8.9 g/dL (range, 8.4-9.1 g/dL). The Starting DA dose was 75 microg per week in 3.7% of patients, 150 microg per week in 65.4% of patients, and 300 microg per week in 29.6% of patients (the dose was increased in 18.5% of patients and reduced in 9.9% of patients; median time to dose adjustment, 8 weeks). Five patients received granulocyte colony stimulating factors. No DA-related adverse reactions occurred. CONCLUSIONS: In the current study, 55% of evaluable patients with MDS safely achieved an erythroid response. PMID- 17115425 TI - Genome-based identification and characterization of a putative mucin-binding protein from the surface of Streptococcus pneumoniae. AB - Streptococcus pneumoniae open reading frame SP1492 encodes a surface protein that contains a novel conserved domain similar to the repeated fragments of mucin binding proteins from lactobacilli and lactococci. To investigate the functional role(s) of this protein and its potential adhesive properties, the surface exposed region of SP1492 was expressed in Escherichia coli, purified to homogeneity, and partially characterized by biophysical and immunological methods. Circular dichroism and sedimentation measurements confirmed that SP1492 is an all-beta protein that exists in solution as a monomer. The SP1492 protein has been shown to be expressed by S. pneumoniae and was experimentally localized to its surface. The protein functional domain binds to mucins II and III from porcine stomach and to purified submaxillary bovine gland mucin. It appears to be one of the very few unambiguous pneumococcal adhesin molecules known to date. A hypothetical model constructed by ab initio techniques predicts a novel beta sandwich protein structure. PMID- 17115426 TI - Molecular dynamics studies of hexamers of amyloid-beta peptide (16-35) and its mutants: influence of charge states on amyloid formation. AB - To study the early stage of amyloid-beta peptide (Abeta) aggregation, hexamers of the wild-type (WT) Abeta(16-35) and its mutants with amyloid-like conformations have been studied by molecular dynamics simulations in explicit water for a total time of 1.7 micros. We found that the amyloid-like structures in the WT oligomers are destabilized by the solvation of ionic D23/K28 residues, which are buried in the fibrils. This means that the desolvation of D23/K28 residues may contribute to the kinetic barrier of aggregation in the early stage. In the E22Q/D23N, D23N/K28Q, and E22Q/D23N/K28Q mutants, hydration becomes much less significant because the mutated residues have neutral amide side-chains. These amide side chains can form linear cross-strand hydrogen bond chains, or "polar zippers", if dehydrated. These "polar zippers" increase the stability of the amyloid-like conformation, reducing the barrier for the early-stage oligomerization. This is in accord with experimental observations that both the D23/K28 lactamization and the E22Q/D23N mutation promote aggregation. We also found that the E22Q/D23N mutant prefers an amyloid-like conformation that differs from the one found for WT Abeta. This suggests that different amyloid structures may be formed under different conditions. PMID- 17115427 TI - Interpersonal process and outcome in variants of cognitive-behavioral psychotherapy. AB - Early sessions from three variants of cognitive-behavioral therapy (CBT) were examined to replicate work done in psychodynamic-interpersonal treatments linking interpersonal process to outcome (W. P. Henry, T. E. Schacht, & H. H. Strupp, 1986, 1990). Cases were available from a component study of CBT for generalized anxiety disorder (T. D. Borkovec, M. G. Newman, A. L. Pincus, & R. Lytle, 2002) and were selected to form good and poor outcome groups maintained through a 1 year follow-up. A third group was also examined that had initial positive outcomes and marked decline by follow-up (n = 8 for each). Structural analysis of social behavior (SASB) was used to identify interpersonal behaviors. Contrary to the authors' expectation, SASB variables were not strong predictors of outcome, and lower levels of interpersonal hostility were found than was the case in previous work. Findings are discussed in light of differences observed between treatment variants and the role that manuals may have in standardizing some aspects of the therapeutic relationship. PMID- 17115428 TI - A longitudinal study of perceived parental psychological control and psychological well-being in Chinese adolescents in Hong Kong. AB - On two occasions separated by one year, Chinese adolescents (N = 2,758) responded to instruments measuring their perceived parental psychological control and psychological well-being, including hopelessness, mastery, life satisfaction, and self-esteem. Pearson correlation analyses revealed that perceived parental psychological control was concurrently related to adolescent psychological well being at Time 1 and Time 2. Multiple regression analyses demonstrated that the relationships between perceived parental psychological control and adolescent psychological well-being over time were bidirectional in nature. Regarding the differential contribution of paternal and maternal psychological control to adolescent psychological well-being over time, paternal psychological control at Time 1 predicted changes in adolescent life satisfaction at Time 2, particularly for adolescent girls. On the other hand, maternal psychological control at Time 1 predicted changes in adolescent self-esteem at Time 2. Relative to those conditions in which one or none of the adolescents' parents was perceived to display high psychological control at Time 1, the psychological well-being of adolescents at Time 2 was poorer under the condition in which both parents were perceived to display high levels of psychological control at Time 1. The clinical implications of these findings and directions for future research are discussed. PMID- 17115429 TI - Examination of the analytic quality of behavioral health randomized clinical trials. AB - Adoption of evidence-based practice (EBP) policy has implications for clinicians and researchers alike. In fields that have already adopted EBP, evidence-based practice guidelines derive from systematic reviews of research evidence. Ultimately, such guidelines serve as tools used by practitioners. Systematic reviews of treatment efficacy and effectiveness reserve their strongest endorsements for treatments that are supported by high-quality randomized clinical trials (RCTs). It is unknown how well RCTs reported in behavioral science journals fare compared to quality standards set forth in fields that pioneered the evidence-based movement. We compared analytic quality features of all behavioral health RCTs (n = 73) published in three leading behavioral journals and two leading medical journals between January 2000 and July 2003. A behavioral health trial was operationalized as one employing a behavioral treatment modality to prevent or treat an acute or chronic physical disease or condition. Findings revealed areas of weakness in analytic aspects of the behavioral health RCTs reported in both sets of journals. Weaknesses were more pronounced in behavioral journals. The authors offer recommendations for improving the analytic quality of behavioral health RCTs to ensure that evidence about behavioral treatments is highly weighted in systematic reviews. PMID- 17115430 TI - Bipolar disorder: improving diagnosis and optimizing integrated care. AB - Bipolar disorder is a chronic, severe condition commonly causing substantial mortality and psychosocial morbidity. Challenges in recognition can delay the institution of appropriate management, whereas misdiagnosis may initiate pharmacologic interventions that adversely affect the condition's course. Pharmacotherapy remains the foundation of treatment. In addition to efficacy, tolerability is an important consideration in medication choice, particularly for long-term maintenance because of its impact on adherence. Mood stabilizers are the classic treatments for bipolar disorder. Newer agents such as atypical antipsychotics may offer efficacy and/or tolerability advantages compared with other medications. The role of antidepressants in bipolar disorder remains controversial. Growing evidence indicates that adjunctive psychosocial interventions improve long-term functioning; consequently, psychologists are becoming increasingly involved in the long-term care of patients with bipolar disorder. This review seeks to update psychologists and related healthcare professionals on recent advances and the current limitations in the diagnosis and treatment of bipolar disorder. PMID- 17115431 TI - The efficacy of personal construct therapy: a comprehensive review. AB - Prior narrative and quantitative reviews have suggested that personal construct therapy (PCT) is an efficacious and viable form of treatment. However, these reviews failed to include all published, controlled PCT outcome studies and did not account for all measures of outcome. The present quantitative review, therefore, offers a comprehensive integration of the 22 published studies that compare PCT to a control group, examining its overall efficacy at posttest and follow-up as well as its effects for different types of outcome measures. Overall, the efficacy of PCT was found to be somewhat weaker than that reported in previous reviews. It was also found that traditional self-report and behavioral observation measures tended to yield larger effects compared to measures that focused on personal meanings and scored content supplied by the client. PMID- 17115432 TI - Relative impact of adverse events and screened symptoms of posttraumatic stress disorder and depression among active duty soldiers seeking mental health care. AB - Symptoms of depression and posttraumatic stress are among the most studied psychological difficulties among soldiers. Such symptoms have been linked to a history of adverse events among both civilians and combat veterans. There is a paucity of research on this topic that can be applied to an active duty clinical population. Intake screening data were reviewed for 1,626 soldiers presenting to an outpatient mental health clinic to identify variables, including history of potentially traumatic experiences, associated with screened symptoms of posttraumatic stress disorder (PTSD) and depression. Demographics such as age, gender, and military rank, as well as number of adverse childhood experiences were significant predictors of screened PTSD and depression. A history of deployment to a combat zone predicted screened PTSD, but not depression. The role of childhood abuse as a risk factor is discussed and highlighted in the etiology of symptoms for soldiers seeking mental health care. PMID- 17115434 TI - Mass-forming extramedullary hematopoiesis diagnosed by fine-needle aspiration cytology. AB - Extramedullary hematopoiesis (EMH) is usually a microscopic finding. However, it may present as a mass-forming lesion making it amenable to fine-needle aspiration biopsy (FNAB). When mass-forming EMH occurs, it can simulate a neoplasm clinically and radiologically. Additionally, the megakaryocytes can mimic malignant neoplastic cells, particularly if EMH is not a considered diagnosis. We report six cases of mass-forming EMH diagnosed by FNAB and evaluate the utility of FNAB in diagnosing EMH. Four patients had prior diagnoses of hematologic disorders, one patient had malignant mastocytosis who presented with lymphadenopathy and one patient had a history of carcinoma. The patients' ages ranged from 46 to 78 yr with an equal sex distribution. Aspirate smears showed trilineage hematopoiesis. The cytomorphologic differential diagnosis included metastatic carcinoma, Hodgkin lymphoma and myeloid sarcoma. No special stains were necessary due to the classic cytologic findings and prior hematologic history. PMID- 17115435 TI - Fine-needle aspiration biopsy for the primary diagnosis of lymphoproliferative disorders involving the spleen: one institution's experience and review of the literature. AB - We report the fine-needle aspiration (FNA) biopsy findings for the primary diagnosis of lymphoproliferative disorders involving the spleen. We analyzed six cases of primary diagnosed lymphoma involving the spleen and out of these cases, identified one case of primary splenic lymphoma. We explore the potential pitfalls and difficulties encountered in making a primary diagnosis of lymphoma involving the spleen and how the preparation of the specimen for flow cytometric studies and/or cell block for immunohistochemical analysis can greatly aid in making a definitive diagnosis. To the best of our knowledge, this is the first report of T-cell rich B-cell subtype of diffuse large B-cell lymphoma and only the second report of primary splenic lymphoma of the spleen diagnosed by FNA biopsy. An extensive review and analysis of the literature involving FNA biopsy of the spleen is also discussed. PMID- 17115436 TI - Positive predictive value of the breast FNAB diagnoses of epithelial hyperplasia with atypia, papilloma, and radial scar. AB - Proliferative breast lesions are a spectrum of lesions that have overlapping cytological features and are difficult to subclassify in fine-needle aspiration biopsies (FNABs). This study of breast FNABs from the period of 1997-2001 aimed to correlate the cytological diagnoses of epithelial hyperplasia with atypia (EHA), papilloma (PAP), and radial scar/complexing sclerosing lesions with relevant histological follow-up, and to assess the positive predictive value (PPV) of the cytological diagnosis. The PPV for the three categories were found to be 0.16, 0.74, and 0.63 respectively. The low value observed in the cytological category of EHA was due to the heterogeneous nature of the group, which makes it difficult to correlate cytological appearances with the varying histological lesions. However, it was found that 70% of these EHA cases had benign histological diagnoses, matching the expected outcome. The PPV of 0.74 for the cytological diagnosis of PAP of the breast compares favorably to similar published studies, and we suggest that the specific diagnosis of PAP can be made reliably using cytological criteria. PMID- 17115437 TI - Seminal vesicle cell in a spontaneously voided urine. PMID- 17115438 TI - Lymph node metastasis of large-cell carcinoma of the lung in a seventeen-year-old patient: diagnosis by fine-needle aspiration. PMID- 17115439 TI - The diagnostic utility of D2-40 for malignant mesothelioma versus pulmonary carcinoma with pleural involvement. AB - Differentiating malignant mesothelioma (MM) from pulmonary carcinoma in pleural fluid cytology can be challenging. Recent studies have suggested that D2-40, a novel lymphatic marker, may be a useful marker for mesothelial differentiation in surgical specimens. However, there are no available data regarding its utility in effusion cytology specimens. We investigated the utility of D2-40 in pleural fluid cytology in differentiating MM from pulmonary carcinomas. Twenty cases of pleural effusion smears of surgically confirmed MM with their corresponding cell blocks were retrieved from the database of the hospital computer system. We also included 10 cases of metastatic pulmonary adenocarcinoma (PA) and 10 cases metastatic pulmonary squamous cell carcinoma (PSCC) involving the pleural fluid. Cell blocks were formalin-fixed, paraffin embedded, and immunostained for TTF1, p63, calretinin, CK5/6, WT-1, and D2-40. Cases were scored as negative (<5% positivity) or positive (>5% moderate/strong positivity). The positive rates for TTF1, p63, calretinin, CK5/6, WT-1, and D2-40 were as follows: MM (0/20), (0/20), (17/20), (18/20), (19/20), (17/20), for PA (8/10), (0/10), (3/10), (0/10), (0/10), (0/10), and for PSCC (1/10), (10/10), (6/10), (10/10), (0/15), (0/10). The staining pattern for D2-40 was characterized by thick membranous staining. Diffuse cytoplasmic staining by D2-40 was seen in 2 cases of pulmonary carcinoma, counted as negative. Our study showed that in differentiating MM from PA, CK5/6, WT-1, and D2-40 have high specificity and sensitivity for MM. Although calretinin is a sensitive IHC marker for MM, it is not specific since it stained 30% of PA. Conversely, to differentiate between MM and PSCC, p63 and WT-1 are the best available markers. We recommend a panel of CK5/6, p63, D2-40, and WT-1 to differentiate MM from pulmonary carcinomas in effusion cytology specimens. PMID- 17115440 TI - The role of fine-needle aspiration cytology and Ziehl Neelsen staining in the diagnosis of cutaneous tuberculosis. AB - The present study highlights the role of fine-needle aspiration cytology (FNAC) and Ziehl Neelsen (ZN) staining in diagnosis of cutaneous tuberculosis and correlates the cytomorphological features with histopathology.FNAC and biopsy was performed on 30 cases of cutaneous tuberculosis and along with the routine stains, ZN and periodic acid Schiffs staining was carried out in all cases. On cytology, out of 9 cases of lupus vulgaris, 89% showed cohesive epithelioid cell granulomas with or without chronic inflammatory infiltrate; however, acid fast bacilli (AFB) could be demonstrated only in 22.2% on cytology while none on histopathology. Of 19 cases diagnosed as Scrofuloderma, 79% showed caseation necrosis with or without granulomas, 10.5% revealed granulomas with acute inflammatory infiltrates. AFB was demonstrated in 78.9% cases on cytology when compared with 15.8% on histopathology. No conclusion could be drawn in one case each of TBVC and lichen scrofulosorum.Hence, correlating cytomorphological patterns with clinical presentations often yields diagnostic information in cases of cutaneous tuberculosis and frequently obviates the need for biopsy especially in cases of scrofuloderma. PMID- 17115441 TI - Fine-needle aspiration cytology of papillary renal cell carcinoma: the association with concomitant secondary malignancies. AB - Papillary renal cell carcinoma is a rare type of renal malignancy. Cytogenetic findings characteristic for this tumor have been described as well as mutations of the proto-oncogene c-met. Secondary malignancies occurring together with papillary renal cell carcinomas are rare, and are often of genitourinary tract origin. We describe two cases of papillary renal cell carcinoma occurring in association with two other visceral malignancies, gastrointestinal stromal tumor and colon adenocarcinoma.Two cases of papillary renal cell carcinoma diagnosed by fine-needle aspiration (FNA), occurring in association with gastrointestinal malignancies were reviewed. Both aspirates showed cytologic features characteristic for papillary renal cell carcinoma, namely papillary structures, foamy histiocytes, intracytoplasmic hemosiderin, and nuclear grooves. Subsequent histology and immunohistochemical stains supported the cytologic diagnosis. The histologic diagnosis of gastrointestinal stromal tumor and colon adenocarcinoma were confirmed. Papillary renal cell carcinoma is a type of renal carcinoma that can be often accurately diagnosed by FNA. The occurrence of associated visceral malignancies has never been reported. The possible role of the protooncogene c met in the development of these tumors was explored. PMID- 17115442 TI - Fine-needle aspiration of a paraesophageal diverticulum masquerading as a thyroid nodule. AB - Fine-needle aspiration (FNA) is a widely utilized cost-effective method for the initial evaluation and triage of thyroid nodules. On occasion, a variety of non thyroidal lesions can be mistaken for thyroid nodules on physical examination or in imaging studies and aspirated. Paraesophageal/ pharyngoesophageal diverticula masquerading as a thyroid nodule have been reported in the radiology literature but, as far as we know, not in the cytology literature. We describe FNA of a paraesophageal diverticulum that had been reported as a thyroid nodule on ultrasound. Cytologists should be familiar with this entity and suggest the diagnosis when appropriate. PMID- 17115443 TI - Reed-Sternberg cells in bronchial brushings from a patient with Hodgkin's disease. PMID- 17115444 TI - Variation of baculovirus-harbored transgene transcription among mesenchymal stem cell-derived progenitors leads to varied expression. AB - We have previously demonstrated that baculovirus can efficiently transduce human mesenchymal stem cells (MSCs) and MSCs-derived adipogenic, chondrogenic, and osteogenic progenitors without compromising the differentiation capacity. Remarkably, the transgene expression level and duration varied widely with the differentiation states at which the progenitors were transduced. However, whether the variation was a general phenomenon and what caused the variation were unclear. Here we demonstrated that transduction of the MSCs and MSC-derived progenitors using baculoviruses carrying egfp driven by CMV, EF-1alpha or CAG promoter resulted in a general trend of varied expression, that is, the chondrogenic progenitors allowed for the poorest expression while the adipogenic progenitors conferred the best expression. Quantification of the nuclear and cytoplasmic egfp gene copy numbers by quantitative real-time PCR revealed that the varied expression did not arise from the discrepancies in gene delivery efficiency nor was it due to the disparities in nuclear transport efficiency. In contrast, the transcription levels paralleled the overall expression levels. These data suggested that although the egfp genes could be efficiently delivered into the nuclei of chondrogenic progenitors, they were not transcribed as well as they were in the adipogenic progenitors. In conclusion, the rapidly altering cellular transcription machinery in the course of differentiation progression predominantly led to the varied expression levels. PMID- 17115445 TI - Dynamic analysis of GS-NS0 cells producing a recombinant monoclonal antibody during fed-batch culture. AB - In this study we have analyzed the dynamic covariation of the mammalian cell proteome with respect to functional phenotype during fed-batch culture of NS0 murine myeloma cells producing a recombinant IgG(4) monoclonal antibody. GS-NS0 cells were cultured in duplicate 10 L bioreactors (36.5 degrees C, 15% DOT, pH 7.0) for 335 h and supplemented with a continuous feed stream after 120 h. Cell specific growth rate declined continuously after 72 h of culture. Cell-specific recombinant monoclonal antibody production rate (qP) varied sixfold through culture. Whilst qP correlated with relative recombinant heavy chain mRNA abundance up to 216 h, qP subsequently declined, independent of recombinant heavy chain or light chain mRNA abundance. GS-NS0 cultures were sampled at 48 h intervals between 24 and 264 h of culture for proteomic analyses. Total protein abundance and nascent polypeptide synthesis was determined by 2D PAGE of unlabeled proteins visualized by SYPRO Ruby and autoradiography of (35)S-labeled polypeptides, respectively. Covariation of nascent polypeptide synthesis and abundance with biomass-specific cell growth, glucose and glutamate consumption, lactate and Mab production rates were then examined using two partial least squares regression models. Most changes in polypeptide synthesis or abundance for proteins previously identified by mass spectrometry were positively correlated with biomass-specific growth rate. We conclude that the substantial transitions in cell physiology and qP that occur during culture utilize a relatively constant complement of the most abundant host cell machines that vary primarily with respect to induced changes in cell growth rate. PMID- 17115446 TI - Measurement of biodegradability parameters for single unsubstituted and methylated polycyclic aromatic hydrocarbons in liquid bacterial suspensions. AB - Substrate depletion experiments were conducted to characterize aerobic biodegradation of 20 single polycyclic aromatic hydrocarbons (PAHs) by induced Sphingomonas paucimobilis strain EPA505 in liquid suspensions. PAHs consisted of low molecular weight, unsubstituted, and methyl-substituted homologs. A material balance equation containing the Andrews kinetic model, an extension of the Monod model accounting for substrate inhibition, was numerically fitted to batch depletion data to estimate extant kinetic parameters including the maximal specific uptake rates, q(max), the affinity coefficients, K(S), and the substrate inhibition coefficients, K(I). Strain EPA505 degraded all PAHs tested. Applied kinetic models adequately simulated experimental data. A cell proliferation assay involving reduction of the tetrazolium dye WST-1 was used to evaluate the ability of strain EPA505 to utilize individual PAHs as sole energy and carbon sources. Of the 22 PAHs tested, 9 supported bacterial growth. Evaluation of the biokinetic data showed that q(max) correlated highly with transmembrane flux as theoretically estimated by a diffusion model, pointing to transmembrane transport as a potential rate-determining process. The biodegradability data generated in this study is essential for the development of quantitative structure-activity relationships (QSARs) for biodegradability and for modeling biodegradation of simple PAH mixtures. PMID- 17115447 TI - Kinetics of biodegradation of binary and ternary mixtures of PAHs. AB - The kinetics of biodegradation of mixtures of polycyclic aromatic hydrocarbons (PAHs) by Sphingomonas paucimobilis strain EPA505 were investigated. The investigation focused on three- and four-ring PAHs, specifically 2 methylphenanthrene, fluoranthene, and pyrene. Uptake rates in aerobic batch suspended cultivations were measured for the individual PAHs and their binary and ternary mixtures. It was observed that kinetics were influenced by the mixture composition and the kinetic properties of the components. A material balance equation containing the Monod model was numerically fitted to uptake data to determine extant kinetic parameters for the individual PAHs. Similarly, equations containing kinetic interaction models derived from enzyme kinetics were fitted to the uptake data obtained from experiments with binary and ternary mixtures. The investigation considered the following interaction types: no-interaction (Monod), pure competitive interaction, noncompetitive or mixed-type interaction, uncompetitive inhibition, and nonspecific interaction based on pure competition (SKIP). Model fit was evaluated based on probabilistic and statistical criteria and inferences were reached about underlying interaction mechanisms based on model fit. Mixture kinetics were most adequately simulated by the pure competitive interaction model with mutual substrate exclusivity. This model is fully predictive, relying only on parameters determined in the sole-PAH experiments. It was shown that for low percent inhibition values and with limited data, pure competitive interaction kinetics may not be evident, resembling no interaction kinetics. This study is a reasonable starting point for understanding and modeling biodegradation of complex PAH mixtures in engineered and natural systems. PMID- 17115448 TI - Nitrifying bacterial communities in an aquaculture wastewater treatment system using fluorescence in situ hybridization (FISH), 16S rRNA gene cloning, and phylogenetic analysis. AB - Aquaculture, especially shrimp farming, has played a major role in the growth of Thailand's economy in recent years, as well as in many South East Asian countries. However, the nutrient discharges from these activities have caused adverse impacts on the quality of the receiving waterways. In particular nitrogenous compounds, which may accumulate in aquaculture ponds, can be toxic to aquatic animals and cause environmental problems such as eutrophication. The mineralization process is well known, but certain aspects of the microbial ecology of nitrifiers, the microorganisms that convert ammonia to nitrate, are poorly understood. A previously reported enrichment of nitrifying bacteria (ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB)) from a shrimp farm inoculated in a sequencing batch reactor (SBR) was studied by molecular methods. The initial identification and partial quantification of the nitrifying bacteria (AOB and NOB) were carried out by fluorescence in situ hybridization (FISH) using previously published 16S rRNA-targeting oligonucleotide probes. The two dominant bacterial groups detected by FISH were from the Cytophaga-Flavobacterium-Bacteroides and Proteobacteria (beta subdivision) phyla. Published FISH probes for Nitrobacter and Nitrospira did not hybridize to any of the bacterial cells. Therefore it is likely that new communities of NOBs, differing from previously reported ones, exist in the enrichments. Molecular genetic techniques (cloning, sequencing, and phylogenetic analysis) targeting the 16S rRNA genes from the nitrifying enrichments were performed to identify putative AOBs and NOBs. PMID- 17115449 TI - A simplified bioprocess for human alpha-fetoprotein production from inclusion bodies. AB - A simple and effective Escherichia coli (E. coli) bioprocess is demonstrated for the preparation of recombinant human alpha-fetoprotein (rhAFP), a pharmaceutically promising protein that has important immunomodulatory functions. The new rhAFP process employs only unit operations that are easy to scale and validate, and reduces the complexity embedded in existing inclusion body processing methods. A key requirement in the establishment of this process was the attainment of high purity rhAFP prior to protein refolding because (i) rhAFP binds easily to hydrophobic contaminants once refolded, and (ii) rhAFP aggregates during renaturation, in a contaminant- dependent way. In this work, direct protein extraction from cell suspension was coupled with a DNA precipitation centrifugation step prior to purification using two simple chromatographic steps. Refolding was conducted using a single-step, redox-optimized dilution refolding protocol, with refolding success determined by reversed phase HPLC analysis, ELISA, and circular dichroism spectroscopy. Quantitation of DNA and protein contaminant loads after each unit operation showed that contaminant levels were reduced to levels comparable to traditional flowsheets. Protein microchemical modification due to carbamylation in this urea-based process was identified and minimized, yielding a final refolded and purified product that was significantly purified from carbamylated variants. Importantly, this work conclusively demonstrates, for the first time, that a chemical extraction process can substitute the more complex traditional inclusion body processing flowsheet, without compromising product purity and yield. This highly intensified and simplified process is expected to be of general utility for the preparation of other therapeutic candidates expressed as inclusion bodies. PMID- 17115450 TI - Regulation and characterization of the polarity of cells embedded in a reconstructed basement matrix using a three-dimensional micro-culture system. AB - Three cell lines, that is, the human breast cancer cell line (MCF-7) and the human mammary epithelial cell line (S-1) and its malignant form (T4-2) were embedded in a reconstituted basement membrane (Matrigel) that had 20-nL pyramid shaped silicon microstructures. The proliferative behavior of the MCF-7 cells was dependent on the surrounding conditions (2-D, collagen gel, or Matrigel), whereas the respiratory activity of a single cell (F(c)) was almost identical under different culture conditions. The F(c) value changed with cellular polarity. The F(c) value for the S-1 cells was observed to decrease slightly, whereas that of the T4-2 cells increased 2 days after cultivation in the microstructures within the Matrigel. However, when the T4-2 cells were cultured in the presence of tyrphostin AG 1478 (T4-2 tyr) to inhibit epidermal growth factor (EGF) signaling, the F(c) value decreased slightly and remained almost constant for an additional 1 week; this was similar to the behavior of the S-1 cells. Further, fluorescence images showed that the T4-2 tyr cells formed polar structures that were similar to those formed by the S-1 cells whereas the T4-2 cells did not form such structures. These results indicate that cellular polarity can be assessed by measuring cellular respiratory activity. PMID- 17115451 TI - Degradation of supercoiled plasmid DNA within a capillary device. AB - Supercoiled plasmid DNA is susceptible to fluid stress in large-scale manufacturing processes. A capillary device was used to generate controlled shear conditions and the effects of different stresses on plasmid DNA structure were investigated. Computational fluid dynamics (CFD) analysis was employed to characterize the flow environment in the capillary device and different analytical techniques were used to quantify the DNA breakage. It was found that the degradation of plasmid DNA occurred at the entrance of the capillary and that the shear stress within the capillary did not affect the DNA structure. The degradation rate of plasmids was well correlated with the average elongational strain rate or the pressure drop at the entrance region. The conclusion may also be drawn that laminar shear stress does not play a significant role in plasmid DNA degradation. PMID- 17115452 TI - High O2 affinity hemoglobin-based oxygen carriers synthesized via polymerization of hemoglobin with ring-opened 2-chloroethyl-beta-D-fructopyranoside and 1-o octyl-beta-D-glucopyranoside. AB - Second generation hemoglobin-based O(2) carriers (HBOCs) are being developed with high O(2) affinity (low P(50)) in order to suppress vasoconstriction elicited by over-oxygenating tissues, a problem associated with low O(2) affinity first generation HBOCs. Our group has previously investigated the polymerization of hemoglobin (Hb) with dialdehydes as a strategy for engineering high O(2) affinity HBOCs. In this study, two novel reactive dialdehydes were synthesized by ring opening 2-chloroethyl-beta-D-fructopyranoside (2-CEFP) and 1-o-octyl-beta-D glucopyranoside (1-OGP) at the 1,2-diol position, respectively, to yield novel Hb polymerizing reagents. High-affinity polymerized HBOCs were synthesized by reacting R-state bovine hemoglobin (bHb) with ring-opened 2-CEFP and 1-OGP at cross-linker to bHb molar ratios ranging from 10:1 to 30:1. The resulting polymerized bovine HBOCs (bHBOCs) displayed P(50)s ranging from 7 to 18 mmHg, cooperativities ranging from 0.8 to 1.4, and methemoglobin (metHb) levels ranging from 3% to 10%. The cross-linking reaction also stabilized the third stepwise Adair coefficient for bHbs reacted with ring-opened 1-OGP at cross-linker to bHb molar ratios of 20:1 and 30:1 and for bHbs reacted with ring-opened 2-CEFP at molar ratios of 30:1. Additionally, the number-averaged molecular weight, M(n), of each polymerized bHBOC was larger compared to bHb. Molecular weight distributions leaning towards larger molecular weight bHBOCs were obtained by increasing the cross-linker to bHb molar ratio. Taken together, the results of this study have identified novel Hb polymerization reagents that are easy to synthesize, and that are capable of yielding bHBOCs with higher O(2) affinities and weight-averaged molecular weights compared to bHb. PMID- 17115453 TI - Proteomic analysis of log to stationary growth phase Lactobacillus plantarum cells and a 2-DE database. AB - Lactobacillus plantarum is part of the natural microbiota of many food fermentations as well as the human gastro-intestinal tract. The cytosolic fraction of the proteome of L. plantarum WCFS1, whose genome has been sequenced, was studied. 2-DE was used to investigate the proteins from the cytosolic fraction isolated from mid- and late-log, early- and late-stationary phase cells to generate reference maps of different growth conditions offering more knowledge of the metabolic behavior of this bacterium. From this fraction, a total of 200 protein spots were identified by MALDI-MS and a proteome production map was constructed to facilitate further studies such as detection of suitable biomarkers for specific growth conditions. More than half (57%) of the identified proteins were predicted to be involved in metabolic pathways of the bacterium. The protein profile changed during the growth of the bacteria such that 29% of the identified proteins involved in anabolic pathways were at least twofold up regulated throughout the mid- and late-exponential and early-stationary phases. In the late-stationary phase, six proteins involved in stress or with a potential role for survival during starvation were up-regulated significantly. PMID- 17115454 TI - Three stage contingent screening for Down syndrome. PMID- 17115456 TI - De novo chromosomal abnormalities and month of conception. Data from the southern hemisphere. PMID- 17115457 TI - Limited value of echography to predict true fetal mosaicism for trisomy 12. PMID- 17115458 TI - Psychological distress and concerns of elderly patients treated with palliative radiotherapy for lung cancer. AB - We conducted a prospective observational cohort study of 83 elderly patients (aged 75 and above) being treated with palliative radiotherapy for lung cancer, with a comparison group of 49 younger patients (aged 65 and under). Psychological distress and concerns were measured before and after treatment using the Hospital Anxiety and Depression Scale (HADS) and a Concerns Checklist. Psychosocial morbidity was common, however, prevalence was similar in both age groups. There was a trend towards worsening of both anxiety and depression scores after treatment, but this did not reach statistical significance. Younger patients reported more concerns than the older group (median 12 vs 10) but this too was not statistically significant. Concerns about the illness and symptoms were more likely to have been addressed by the care team than were concerns about psychosocial issues such as the family and the future. People of all ages have similar concerns and levels of anxiety and depression whilst receiving palliative radiotherapy for lung cancer. Further research is needed to explore the use of screening tools, like those used in this study, to identify patients' difficulties and target interventions to improve their quality of life. PMID- 17115459 TI - Complex biohopanoids synthesis: efficient anchoring of ribosyl subunits onto a C(30) hopane. AB - Bacteriohopanoids represent a particularly important series of triterpenoids, widely distributed in bacteria. One of the common features of these pentacyclic hopanepolyols is the presence of an extended non-terpenoid and polyhydroxylated side chain attached to the triterpenic moiety through a C--C bond. The biological function of biohopanoids also has to be addressed when one considers the broad diversity in both structures and functionalities found in the side chain. Moreover, the stereochemistries of some biohopanoids are still unconfirmed, due to the lack of synthetic methods to prepare them. In this study we describe an efficient methodology for the formation of the C--C bond between the C(30)-hopane component and C(5)-polyhydroxylated carbohydrates through the use of a hopanyllithium intermediate, which has enabled us to synthesize several biohopanoid derivatives. We also report the first synthesis of hopanepentol bearing an additional hydroxy group at position C31. PMID- 17115460 TI - Oxo- and imidovanadium complexes incorporating methylene- and dimethyleneoxa bridged calix[3]- and -[4]arenes: synthesis, structures and ethylene polymerisation catalysis. AB - Reaction of [V(X)(OR)3] (X=O, Np-tolyl; R=Et, nPr or tBu) with p-tert butylhexahomotrioxacalix[3]areneH3, LH3, affords the air-stable complexes [{V(X)L}n] (X=O, n=1 (1); X=Np-tolyl, n=2 (2)). Alternatively, 1 is readily available either from interaction of [V(mes)3THF] with LH3, and subsequent oxidation with O2 or upon reaction of LLi3 with [VOCl3]. Reaction of [V(Np tolyl)(OtBu)3] with 1,3-dimethylether-p-tert-butylcalix[4]areneH2, Cax(OMe)2(OH)2, afforded [{VO(OtBu)}2(mu-O)Cax(OMe)2(O)2].2 MeCN (42 MeCN), in which two vanadium atoms are bound to just one calix[4]arene ligand; the n propoxide analogue of 4, namely [{VO(OnPr)}2(mu-O)Cax(OMe)2(O)2].1.5 MeCN (51.5 MeCN), has also been isolated from a similar reaction using [V(O)(OnPr)3]. Reaction of [VOCl3], LiOtBu, (Me3Si)2O and Cax(OMe)2(OH)2 gave [{VO(OtBu)Cax(OMe)2(O)2}2Li4O2].8 MeCN (68 MeCN), in which an Li4O4 cube (two of the oxygen atoms are derived from the calixarene ligands) is sandwiched between two Cax(OMe)2(O)2. The reaction between [V(Np-tolyl)(OtBu)3] and Cax(OMe)2(OH)2, afforded [V(Np-tolyl)(OtBu)2Cax(OMe)2(O)(OH)]5 MeCN (75 MeCN), in which two tert butoxide groups remain bound to the tetrahedral vanadium atom, which itself is bound to the calix[4]arene through only one phenolic oxygen atom. Reaction of p tert-butylcalix[4]areneH4, Cax(OH)4 and [V(Np-tolyl)(OnPr)3] led to loss of the imido group and formation of the dimeric complex [{VCax(O)4(NCMe)}2].6 MeCN (86 MeCN). Monomeric vanadyl oxo- and imidocalix[4]arene complexes [V(X)Cax(O)3(OMe)(NCMe)] (X=O (11), Np-tolyl (12)) were obtained by the reaction of the methylether-p-tert-butylcalix[4]areneH3, Cax(OMe)(OH)3, and [V(X)(OR)3] (R=Et or nPr). Vanadyl calix[4]arene fragments can be linked by the reaction of 2,6-bis(bromomethyl)pyridine with Cax(OH)4 and subsequent treatment with [VOCl3] to afford the complex [{VOCax(O)4}2(mu-2,6-(CH2)2C5H3N)].4 MeCN (134 MeCN). The compounds 1-13 have been structurally characterised by single-crystal X-ray diffraction. Upon activation with methylaluminoxane, these complexes displayed poor activities, however, the use of dimethylaluminium chloride and the reactivator ethyltrichloroacetate generates highly active, thermally stable catalysts for the conversion of ethylene to, at 25 degrees C, ultra-high molecular-weight (>5, 500,000), linear polyethylene, whilst at higher temperature (80 degrees C), the molecular weight of the polyethylene drops to about 450,000. Using 1 and 2 at 25 degrees C for ethylene/propylene co-polymerisation (50:50 feed) leads to ultra-high-molecular-weight (>2,900,000) polymer with about 14.5 mol% propylene incorporation. The catalytic systems employing the methyleneoxa bridged complexes 1 and 2 are an order of magnitude more active than the bimetallic complexes 5 and 13, which, in turn, are an order of magnitude more active than pro-catalysts 8, 11 and 12. These differences in activity are discussed in terms of the structures of each class of complex. PMID- 17115461 TI - Identification of 7,4'-dihydroxy-5-methoxyflavylium in "Dragon's blood": to be or not to be an anthocyanin. AB - The compound 7,4'-dihydroxy-5-methoxyflavylium (dracoflavylium) was identified as the major red colorant in samples of the resin "dragon's blood", extracted from the tree Dracaena draco. The complex network of reversible chemical reactions that dracoflavylium undergoes in aqueous solution is fully described; for the first time, all the equilibrium constants that enable a complete characterisation of the system have been obtained (K'(a)=1.6 x 10(-4), K(a1)=1.0 x 10(-4), K(a2)=3.2 x 10(-8), K(Ct1)=1.0 x 10(-7), K(Ct2)=1.3 x 10(-10)). It is concluded that the red colour is due to a stable quinoid base, A, which is the major species at pH 4-7. It is further shown that this compound does not fit the commonly accepted definitions of anthocyanidin nor 3-deoxyanthocyanidin. Similarly to synthetic flavylium salts, the natural compound 7,4'-dihydroxy-5 methoxyflavylium gives rise to several species (multistate system) reversibly interconverted by external stimuli, such as pH. PMID- 17115462 TI - Long-range-distance NMR effects in a protein labeled with a lanthanide-DOTA chelate. AB - A two-thiol reactive lanthanide-DOTA (1,4,7,10-tetraazacyclododecane N,N',N'',N'''-tetraacetic acid) chelate, CLaNP-3 (CLaNP=caged lanthanide NMR probe), was synthesized for the rigid attachment to cysteine groups on a protein surface, and used to obtain long-range-distance information from the {15N,1H} HSQC spectra of the protein-lanthanide complex. The DOTA ring exhibits several isomers that are in exchange; however, single resonances were observed for most amide groups in the protein, allowing determination of a single, apparent magnetic-susceptibility tensor. Pseudocontact shifts caused by Yb-containing CLaNP-3 were observed for atoms at 15-35 A from the metal. By using Gd-containing CLaNP-3, relaxation effects were observed, allowing distances up to 30 A from the paramagnetic center to be determined accurately. Similar results were obtained with a Gd-DTPA (diethylene-triaminepentaacetic acid) chelate, CLaNP-1, bound in the same bidentate manner to the protein. This study demonstrates that bidentate attachment of a paramagnetic probe enables determination of long-range distances. PMID- 17115463 TI - Characterizing ionic liquids as reaction media through a chemical probe. AB - The triplet N,N-dimethylaminophenyl cation, a highly reactive but chemospecific electrophile, has been used as a probe for characterizing the properties of reaction media for a series of imidazolium ILs. With the N-hexyl-N-methyl imidazolium derivatives (not with the N-butyl analogues), hydrogen transfer leading to the aniline was the main process. Trapping by iodide occurred with an inverse dependence on viscosity. Trapping by pi nucleophiles exhibited a more complex behavior. This was explained by the effect of both the bulk viscosity and the structure of the IL cation on both steps of the reaction, namely, initial electrophilic attack and ensuing cation elimination or nucleophile addition. However, with an excellent nucleophile, such as thiophene, or when the latter step was intramolecular, as with 4-pentenol, the difference was obliterated and trapping became uniform. Incorporation of the probe into the IL cation (through insertion into the C--H bond alpha to the imidazolium ring) was demonstrated, while no addition to the anion tested (including bis(trifluoromethanesulfonimide)) took place. PMID- 17115464 TI - Mechanistic insights into stereoselective catalysis-the effects of counterions in a CuII-bissulfoximine-catalyzed Diels-Alder reaction. AB - The initial steps of an enantioselective Diels-Alder reaction catalyzed by a CuII bissulfoximine complex were followed by EXAFS (EXAFS=extended X-ray absorption fine structure), EPR (EPR=electron paramagnetic resonance) spectroscopy (CW-EPR, FID-detected EPR, pulse ENDOR, HYSCORE; CW=continuous wave; ENDOR=electron nuclear double resonance; HYSCORE=hyperfine sublevel correlation; FID=free induction decay), and UV-visible spectroscopy. The complexes formed between the parent CuX2 (X=Cl-, Br-, TfO-, SbF6-) salts, the chiral bissulfoximine ligand (S,S)-1, and N-(1-oxoprop-2-en-1-yl)oxazolidin-2-one (2) as the substrate in CH2Cl2 were investigated in frozen and fluid solution. In all cases, penta- or hexacoordinated CuII centers were established. The complexes with counterions indicating high stereoselectivity (TfO- and SbF6-) reveal one unique species in which substrate 2 binds to pseudoequatorial positions (via O atoms), shifting the counterions to axial locations. On the other hand, those lacking stereoselectivity (X=Cl- and Br-) form two species in which the parent halogen anions remain at equatorial positions preventing the formation of geometries compatible with those found for X=TfO- and SbF6-. PMID- 17115465 TI - Catalytic applications of metal nanoparticles in imidazolium ionic liquids. AB - Metal nanoparticles (MNPs) with a small diameter and narrow size distribution can be prepared by H(2) reduction of metal compounds or decomposition of organometallic species dissolved in ionic liquids (ILs). MNPs dispersed in ILs are catalysts for reactions under multiphase conditions. These soluble MNPs possess a pronounced surfacelike rather than single-site like catalytic properties. In other cases the MNPs are not stable and tend to aggregate or serve as reservoirs of mononuclear catalytically active species. PMID- 17115468 TI - Recent applications of chiral ferrocene ligands in asymmetric catalysis. AB - Despite the impressive progress achieved in asymmetric catalysis during the last decade, an increasing number of new catalysts, ligands, and applications are reported every year to satisfy the need to embrace a wider range of reactions and to improve the efficiency of existing processes. Because of their availability, unique stereochemical aspects, and wide variety of coordination modes and possibilities for the fine-tuning of the steric and electronic properties, ferrocene-based ligands constitute one of the most versatile ligand architectures in the current scenario of asymmetric catalysis. Over the last few years ferrocene catalysts have been successfully applied in an amazing variety of enantioselective processes. This Review documents these recent advances, with special emphasis on the most innovative asymmetric processes and the development of novel, efficient types of ferrocene ligands. PMID- 17115470 TI - Confidence intervals for the standardized effect arising in the comparison of two normal populations. AB - Confidence intervals for a standardized effect are derived after stabilizing the variance of the Welch t-statistic. Simulation studies demonstrate the viability of the resulting intervals for a wide range of parameter values and sample sizes as small as five. The methodology is extended to the combination of results from several studies, so as to obtain a confidence interval for a representative standardized effect for all the studies. The methods are illustrated on a recent meta-analytic study of systolic blood pressure reduction during a weight reducing regime, as well as the classical Mumford data on psychological intervention and hospital length of stay. PMID- 17115471 TI - Blocking breast cancer relapses. PMID- 17115472 TI - When treatment is hard to take. PMID- 17115473 TI - Moving forward after a stroke. PMID- 17115474 TI - Skipping the syringe: options for insulin. PMID- 17115475 TI - Vitamin D: the bone builder. PMID- 17115476 TI - Is coffee unhealthy? Is decaffeinated coffee better for my health? PMID- 17115477 TI - The creation of the almshouse: institutions as solutions to the problem of poverty. PMID- 17115478 TI - Gastrostomy feeding in cerebral palsy: too much of a good thing? PMID- 17115479 TI - Improving, murine health surveillance programs with help of on-site enzyme-linked immunosorbent assay. PMID- 17115480 TI - [Treating mental disorders in the elderly: avant-garde control or... lost cause?]. PMID- 17115481 TI - The HEALTH Act's FDA defense to punitive damages: a gift to drug makers or to the public? PMID- 17115482 TI - Listening to the disabled: end-of-life medical decision making and the never competent. PMID- 17115483 TI - The Commerce Clause and federal abortion law: why progressives might be tempted to embrace federalism. PMID- 17115484 TI - Soy, black cohosh may have some benefit for menopause symptoms. PMID- 17115485 TI - Who said murder? PMID- 17115486 TI - Dicing with death : there's a good chance that the pills your doctor prescribed will do you no good and might even harm you. PMID- 17115487 TI - Report calls for easing rules on research involving prisoners. PMID- 17115488 TI - Bush vetoes bill to loosen policy on stem-cell research. PMID- 17115489 TI - Hyaluronic acid and knee osteoarthritis. PMID- 17115490 TI - Editorial. PMID- 17115491 TI - Ethical research with participants who are deaf. AB - When conducting research with participants who are deaf there is a range of potential issues to be considered: issues relating to language and communication, participants' past experience and culture, and the potential power imbalance between researcher and participant. From an ethical perspective these may compromise informed consent and confidentiality, may inadvertently result in deception and may interfere with the validity of the data. The consequences may be negative for both the deaf population and the research community. This paper highlights the issues that arose in four studies with participants who were deaf and discusses the extent to which the measures taken were effective. PMID- 17115493 TI - Oral as good as intramuscular vitamin B12 replacement. PMID- 17115492 TI - Research in developing countries. AB - The rapid expansion over the last decade in medical research, sponsored by rich countries but undertaken in developing countries, has led to several reports and guidelines, three of which are excerpted here. PMID- 17115494 TI - Methicillin-resistant staph common in abscessed skin infections. PMID- 17115495 TI - Effective methods for preventing pressure ulcers. PMID- 17115496 TI - Human reproductive technologies and the law: a select committee report. AB - The House of Commons Science & Technology Committee has reviewed the Human Fertilisation and Embryology Act. It considered a) the balance between legislation, regulation and reproductive freedom; b) the role of Parliament in human reproductive technologies; and c) the foundation, adequacy and appropriateness of the ethical framework for legislation. It also considered the Act itself and the workings of the Human Fertilisation and Embryology Authority. Its report is written from a very liberal perspective, but is a very thorough overview of current issues and debate in the field. There follow, slightly abridged, the conclusions and recommendations of the 200-page report. PMID- 17115497 TI - Antibiotics slightly effective for purulent rhinitis. PMID- 17115498 TI - The research misconception. AB - Recently, several researchers and philosophers argued that clinical research trials are not therapy. Their position is based on foundational research ethics documents, such as the Belmont Report, on conceptual analysis, and on the general way clinical trials are conducted. After examining and rejecting these arguments, we claim that good research is consistent with good therapy; that often trials are good therapy; and that a blanket attack on clinical trials as non-therapeutic creates a research misconception. This misconception is potentially harmful because it could weaken trial recruitment, could adversely affect funding for trials, and could overturn needed moral safeguards on therapeutic trials. Our more careful and accurate analysis of the nature of clinical trials can avoid such problems. PMID- 17115499 TI - Animal activists flee UK clampdown. PMID- 17115500 TI - The incredibles. PMID- 17115501 TI - Not brain-dead, but ripe for transplant. PMID- 17115502 TI - Man and superman. PMID- 17115503 TI - Essential animals. PMID- 17115504 TI - Big abortion: what the antiabortion movement can learn from big tobacco. PMID- 17115505 TI - An unwelcome discovery. Walter DeNino was a young lab technician who analyzed data for his mentor, Eric Poehlman. What he found was that Poehlman was not the scientist he appeared to be. PMID- 17115506 TI - Neuroscience, nuance, and neuroethics. PMID- 17115507 TI - Supporting organ transplantation in non-resident aliens within limits. AB - It is common knowledge that the supply of cadaveric organs does not meet demand. This shortage is often used as ethical argument against transplantation in Non Resident Aliens; however, this fact in isolation does not present a comprehensive picture of organ allocation in USA. Even though approximately 153 cadaveric livers, kidneys, and hearts are transplanted into Non-Resident Aliens each year, roughly another 85 livers, kidneys and hearts are recovered as usable for transplantation but are not transplanted due to inability to find a recipient. These organs are also unable to be exported due to logistics or lack of patient matching. Because usable, recovered allografts are discarded on a yearly basis, there is no justification to use "allograft scarcity" as argument against transplantation in Non-Resident Aliens. Further, consistent with other countries, a system of two waiting lists which allocates organs to US Residents with the first right of refusal (with Non-Resident Aliens having to access organs refused by or not matched to US Residents) is ethically appropriate. Justification for this two-list system lies in deconstructing "who" is the transplant community, and who are "guests" of the transplant community. PMID- 17115508 TI - The Quebec air medical evacuation program: an evolution influenced by medical progress and airline industry. PMID- 17115509 TI - Possible transmission of hepatitis A in a school setting. PMID- 17115510 TI - Addressing a public health priority: first Canadian research agenda on influenza prevention, control and management. PMID- 17115511 TI - Addendum. Update: guidelines for the prevention and control of meningococcal disease. PMID- 17115512 TI - E. coli O157:H7 outbreak in the United States associated with bagged fresh spinach. PMID- 17115513 TI - Direct-to-consumer online genetic testing and the four principles: an analysis of the ethical issues. AB - The development of genetic tests marketed and sold direct-to-consumers (DTC) via the internet raises moral concerns and debate about their appropriateness and ethical and clinical significance. These tests are offered for a wide range of diseases and conditions, and the mutations have variable penetrance and associated risk. A number of these tests lack data on their accuracy and reliability, making interpretation of results difficult. DTC genetic testing is undertaken outside the context of the physician-patient relationship and may lack appropriate individual and family genetic counseling, leaving the consumer vulnerable to potential harms, such as misinterpretation of results, including false positive or false reassurance, with limited or no benefits. Beauchamp and Childress's four principles of biomedical ethics provide a framework for analyzing the ethical issues raised by DTC genetic testing. We argue that the potential harms outweigh the potential benefits of such tests, that respect for autonomy should be limited in light of potential harm from DTC testing, and that the availability of genetic testing over the internet may be considered unfair and unjust and affect resource allocation by placing an unfair burden on primary care physicians. In light of the moral issues posed by these tests, practical responses are suggested in the areas of consumer education, medical education, and interaction with commercial companies. PMID- 17115514 TI - Worldwide variation in the performance of children and adolescents: an analysis of 109 studies of the 20-m shuttle run test in 37 countries. AB - This study is a meta-analysis of 109 reports of the performance of children and adolescents on the 20-m shuttle run test (20-mSRT). The studies were performed in 37 countries and included data on 418,026 children, tested between 1981 and 2003. Results were expressed as running speed (km x h(-1)) at the final completed stage of the 20-mSRT. Raw data were combined with pseudodata using Monte Carlo simulation. The 20-mSRT performances were expressed as z-scores relative to all children of the same age and sex from all countries. An overall "performance index" was derived for each country as the average of the age- and sex-specific z scores for all children from that country. Factorial analysis of variance was used to compare scores among countries and regions, and between boys and girls of the same age. There was wide and significant (P < 0.0001) global variability in the performance of children. The best performing children were from the Northern European countries Estonia, Iceland, Lithuania, and Finland (0.6 - 0.9 standard deviations above the global average). The worst performing children were from Singapore, Brazil, USA, Italy, Portugal, and Greece (0.4 - 0.9 standard deviations below the global average). There is evidence that performance was negatively related to being overweight, as well as to a country's average temperature. PMID- 17115515 TI - Estimated maturity status and perceptions of adult autonomy support in youth soccer players. AB - In this study, we examined the relations between biological maturity status, body mass index, age, and perceptions of adult autonomy support in the context of youth soccer. A total of 70 female and 43 male soccer players, aged 9 - 15 years, completed three adult-specific versions (i.e. mother, father, coach) of the perceived autonomy support subscale from the Interpersonal Style Scale. The participants' percent predicted adult stature was used as an estimate of biological maturity status. Multiple linear regression analyses revealed that advanced maturity status in male players predicted lower perceptions of autonomy support from the coach. Maturity status was unrelated to perceptions of autonomy support from the coach in female soccer players, and paternal and maternal autonomy support in male and female players. Age and body mass index were unrelated to perceptions of adult (i.e. coach, mother, father) autonomy support in male and female players. PMID- 17115516 TI - Assisted procreation: too little consideration for the babies? AB - Recent studies have revealed much higher risks of cerebral palsy and malformations in babies conceived by in vitro fertilization (IVF) than in babies conceived naturally. Here we question whether parents can legitimately accept this risk on behalf of offspring. We argue that parents can expose their baby to a risk only to preserve it from a worse possibility, and this is not the case of IVF, which is not a therapeutic tool for children because when the IVF decision is taken, the child has not yet been conceived. It is concluded that procreative techniques require considerably more research before being made available to couples. PMID- 17115517 TI - "The Civil Rights Movement of the 1990s?": The anti-abortion movement and the struggle for racial justice. AB - In 1964, Claude and Jeanne Nolen, who were white, joined an interracial NAACP team intent on desegregating local restaurants in Austin, Texas as a test of the recently passed Civil Rights ACt. Twenty-five years later, the Nolens pleaded "no contest" in a courtroom for their continued social activism. This time the issue was not racial segregation, but rather criminal trespassing for blockading abortion clinics with Operation Rescue. The Nolens served prison sentences for direct action protests that they believe stemmed from the same commitment to Christianity and social justice as the civil rights movements. Despite its relationship to political and cultural conservatism, the anti-abortion movement since Roe v. Wade (1973) was also a product of the progressive social movements of the turbulent sixties. Utilizing oral history interviews and organizational literature, the article explores the historical context of the anti-abortion movement, specifically how the lengthy struggle for racial justice shaped the rhetoric, tactics, and ideology of the anti-abortion activists. Even after political conservatives dominated the movement in the 1980s, the successes and failures of the sixties provided a cultural lens through which grassroots anti abortion activists forged what was arguably the largest movement of civil disobedience in American history. PMID- 17115518 TI - Exploring ethical justification for self-demand amputation. AB - Self-demand amputees are persons who need to have one or more healthy limbs or digits amputated to fit the way they see themselves. They want to rid themselves of a limb that they believe does not belong to their body-identity. The obsessive desire to have appendages surgically removed to fit an alternative body-image is medically and ethically controversial. My purpose in this paper is to provide a number of normative and professional ethical perspectives on whether or not it is possible to justify surgery for self-demand amputees. In doing so I proceed dialogically, moving between empirical context and normative theory, revealing the taken for granted normative assumptions (what I call the natural attitude--a technical term borrowed from phenomenology) that provide ethical limits to justifying the treatment of self-demand amputees. While I critically examine both Kantian responses against as well as Utilitarian responses for amputation on demand, I conclude that neither normative tradition can fully incorporate an understanding of what it is like to be a self-demand amputee. Since neither theory can justify the apparent non-rational desire of amputation on demand, ethical justification, I argue, falls short of the recognition that there may be a problem. To end, I introduce a meta-ethical idea, "the struggle for recognition," opening up the theoretical possibility of a hermeneutics of recognition before ethical justification that may be more sensitive to the problem of radical embodied difference exemplified by self-demand amputees. PMID- 17115519 TI - Power output of field-based downhill mountain biking. AB - The purpose of this study was to assess the power output of field-based downhill mountain biking. Seventeen trained male downhill cyclists (age 27.1 +/- 5.1 years) competing nationally performed two timed runs of a measured downhill course. An SRM powermeter was used to simultaneously record power, cadence, and speed. Values were sampled at 1-s intervals. Heart rates were recorded at 5-s intervals using a Polar S710 heart rate monitor. Peak and mean power output were 834 +/- 129 W and 75 +/- 26 W respectively. Mean power accounted for only 9% of peak values. Paradoxically, mean heart rate was 168 +/- 9 beats x min(-1) (89% of age-predicted maximum heart rate). Mean cadence (27 +/- 5 rev x min(-1)) was significantly related to speed (r = 0.51; P < 0.01). Analysis revealed an average of 38 pedal actions per run, with average pedalling periods of 5 s. Power and cadence were not significantly related to run time or any other variable. Our results support the intermittent nature of downhill mountain biking. The poor relationships between power and run time and between cadence and run time suggest they are not essential pre-requisites to downhill mountain biking performance and indicate the importance of riding dynamics to overall performance. PMID- 17115520 TI - The effect of combined self- and expert-modelling on the performance of the double leg circle on the pommel horse. AB - In this study, we investigated whether video modelling can enhance gymnasts' performance of the circle on a pommel horse. The procedure associated expert modelling with self-modelling and quantitative performance analysis. Sixteen gymnasts were randomly assigned to one of two groups: (1) a modelling group, which received expert- and self-modelling, and performance feedback, or (2) a control group, which received no feedback. After five sessions of training, an analysis of variance with repeated measures indicated that the gains in the back, entry, front, and exit phases of the circle were greater for the modelling group than for the control group. During the training sessions, the gymnasts in the modelling group improved their body segmental alignment during the back phase more quickly than during the other phases. As predicted, although both groups performed the same number of circles (300 in 5 days, with 10 sequences of 6 circles), the modelling group improved their body segmental alignment more than the control group. It thus appears that immediate video modelling can help to correct complex sports movements such as the circle performed on the pommel horse. However, its effectiveness seemed to be dependent on the complexity of the phase. PMID- 17115521 TI - When "where" is more important than "when": birthplace and birthdate effects on the achievement of sporting expertise. AB - In this study, we assessed whether contextual factors related to where or when an athlete is born influence their likelihood of playing professional sport. The birthplace and birth month of all American players in the National Hockey League, National Basketball Association, Major League Baseball, and Professional Golfer's Association, and all Canadian players in the National Hockey League were collected from official websites. Monte Carlo simulations were used to verify if the birthplace of these professional athletes deviated in any systematic way from the official census population distribution, and chi-square analyses were conducted to determine whether the players' birth months were evenly distributed throughout the year. Results showed a birthplace bias towards smaller cities, with professional athletes being over-represented in cities of less than 500,000 and under-represented in cities of 500,000 and over. A birth month/relative age effect (in the form of a distinct bias towards elite athletes being relatively older than their peers) was found for hockey and baseball but not for basketball and golf. Comparative analyses suggested that contextual factors associated with place of birth contribute more influentially to the achievement of an elite level of sport performance than does relative age and that these factors are essentially independent in their influences on expertise development. PMID- 17115522 TI - Hormonal and psychological adaptation in elite male rowers during prolonged training. AB - In this study, we examined possible hormonal and psychological changes in elite male rowers during a 24-week preparatory period. Eleven elite male rowers were tested on seven occasions over the 6-month training season. Fasting testosterone, growth hormone, cortisol, and creatine kinase activity, together with perceived recovery-stress state were evaluated after a day of rest. Maximal oxygen consumption (VO2max) was determined before and after the training period. Training was mainly organized as low-intensity prolonged training sessions. Significant increases in VO2max (from 6.2 +/- 0.5 to 6.4 +/- 0.6 l x min(-1)) were observed as a result of training. The overall perceived recovery-stress index did not change during the preparatory period. Standardized recovery and stress scores changed during the course of training in comparison with pre training values. When basal hormone concentrations were compared with the first measurement, significant changes in testosterone and cortisol were observed together with changes in mean weekly training volume. Basal testosterone (r = 0.416; P = 0.010) and cortisol (r = 0.527; P = 0.001) were related to mean weekly training volume. Basal growth hormone did not change during the training. Changes in creatine kinase activity demonstrated similar pattern with changes in mean weekly training volume. The overall perceived recovery-stress index was related to testosterone, cortisol, growth hormone, and creatine kinase activity (r > 0.299; P < 0.015). Our findings indicate that testosterone and cortisol are more sensitive to changes in training volume than either growth hormone or perceived recovery-stress state in elite rowing training. Increases in these stress hormone concentrations represent a positive adaptation to current training load. Significant relationships between hormonal and perceived recovery-stress state suggest that metabolic and psychological changes should be carefully monitored to avoid a negative effect on the training status of elite rowers. PMID- 17115523 TI - Consequences of players' dismissal in professional soccer: a crisis-related analysis of group-size effects. AB - This study documents the effect of players' dismissals on team performance in professional soccer. Our aim was to determine whether the punishment meted out for unacceptable player behaviour results in reduced team performance. The official web site of the German Soccer Association was used for coding data from games played in the first Bundesliga between the 1963 - 64 and 2003 - 04 (n = 41) seasons. A sample of 743 games where at least one red card was issued was used to test hypotheses derived from crisis theory (Bar-Eli & Tenenbaum, 1989a). Players' dismissals weaken a sanctioned team in terms of the goals and final score following the punishment. The chances of a sanctioned team scoring or winning were substantially reduced following the sanction. Most cards were issued in the later stages of matches. The statistics pertaining to outcome results as a function of game standing, game location, and time phases - all strongly support the view that teams can be considered conceptually similar to individuals regarding the link between stress and performance. To further develop the concept of team and individual psychological performance crisis in competition, it is recommended that reversal theory (Apter, 1982) and self-monitoring and distraction theories (Baumeister, 1984) be included in the design of future investigations pertaining to choking under pressure. PMID- 17115524 TI - The effects of synchronous music on 400-m sprint performance. AB - The aim of the present study was to assess the effects of motivating and oudeterous (neither motivating nor demotivating) synchronous music on 400-m sprint performance while controlling for the potential confound of pre performance mood. A panel of volunteer Caucasian males (n = 20; mean age = 20.5 years, s = 1.2) rated the motivational qualities of 32 musical selections using the Brunel Music Rating Inventory-2. An experimental group of volunteer Caucasian males (n = 36; mean age = 20.4 years, s = 1.4) completed three 400-m time trials under conditions of motivational music, oudeterous music, and a no-music control. Pre-performance mood was assessed using the Brunel University Mood Scale (BRUMS). A series of repeated-measures analyses of variance with Bonferroni adjustment revealed no differences in the BRUMS subscales. A repeated-measures analysis of variance on the 400-m times showed a significant effect (F1.24, 42.19 = 10.54, P < 0.001, eta 2 = 0.24) and follow-up pair wise comparisons revealed differences between the synchronous music conditions and the control condition. This finding supported the first research hypothesis, that synchronous music would result in better performance than a no-music control, but not the second hypothesis, that performance in the motivational synchronous music condition would be better than that in the oudeterous condition. It appears that synchronous music can be applied to anaerobic endurance performance among non-elite sports persons with a considerable positive effect. PMID- 17115525 TI - [The League of Nations Health Organization and the rise of Latin American participation, 1920-40]. AB - The League of Nations Health Organization collaborated with Latin American specialists in public health and infectious diseases from the early 1920s to the outbreak of the Second World War. The League developed studies of infant health and nutrition, and leprosy. The approach was expert-oriented, and designed to develop public health on a scientific basis. There were conferences, tours and reports in Latin America. This paper demonstrates that the Latin American collaboration with the Health Organization was extensive and multi-faceted. PMID- 17115526 TI - Thinking outside of the box: the role of environmental adaptation in the acquisition of skilled and expert performance. AB - According to current theories of expert performance, experts gain an advantage by acquiring through practice cognitive skills and strategies that increase the efficiency with which information specific to their domain is processed. Consequently, experts are able to circumvent natural processing limitations. In this study, a description is provided of how experts make use of strategies that involve adapting physical elements of their domain environment to reduce cognitive workload during performance. Telephone interviews were conducted with 15 expert orienteers and six coaches of national orienteering squads about how expert orienteers carry and arrange their navigational equipment while orienteering. A content analysis of the interview data revealed that expert orienteers adapt their navigational equipment to reduce the cognitive and, more specifically, attentional workload during performance. A theory of how experts circumvent natural processing limitations requires consideration of the role of such strategies. PMID- 17115527 TI - ["The demon that turned into worms": the translation of public health in the British Caribbean, 1914-1920]. AB - The earliest programs of the Rockefeller Foundation's International Health Commission - IHC were pilot projects for the treatment of hookworm disease in the British colonies of British Guiana and Trinidad. These pioneering ventures into international health have often been portrayed as governed by rigid biomedical principles. In contrast to this view, the article emphasizes the degree to which the exigencies of a public health project that sought to make biomedicine intelligible within the medical systems of subject populations combined with the knowledge of local IHC staff members of Indo-Caribbean descent to generate some fascinating experiments in ethno-medical translation. One term in particular "The Demon that Turned into Worms" is focused on to show how these efforts at medical translation may have legitimized and promoted medical pluralism. PMID- 17115528 TI - [Pressure or legitimization? Power and alternatives in the planning and adoption of health reforms in Costa Rica, 1988-1998]. AB - Policies are made in response to the rationale of pressure and legitimization, which join forces in many different ways. This work analyzes the planning and adoption of the health reforms undertaken in Costa Rica between 1988 and 1998. It questions whether political parties, international financial institutions and the technical and bureaucratic elites in each sector can be taken as sufficiently explanatory of themselves. Empirical evidence would suggest that apart from investigating the individual interests of the agents involved, one must also consider the sector reforms that are actually available internationally. The paper draws this discussion into the larger scenario of policy making in Latin America and draws links between this and other stages in policy making and other moments in the construction of the State. PMID- 17115529 TI - [The transition from 'international' to 'global' public health and the World Health Organization]. AB - Within the context of international public health, 'global health' seems to be emerging as a recognized term of preference. This article presents a critical analysis of the meaning and importance of 'global health' and situates its growing popularity within a historical context. A specific focus of this work is the role of the World Health Organization - WHO in both 'international' and 'global' health, and as na agent of transition from one to the other. Between 1948 and 1998, the WHO went through a period of hardship as it came up against an organizational crisis, budget cuts and a diminished status, especially when confronted with the growing influence of new, power players like the World Bank. We suggest that the WHO has responded to this changing international context by inititating its own process of restructuring and repositioning as an agent for coordinating, strategically planning and leading 'global health' initiatives. PMID- 17115530 TI - Autologous chondrocyte implantation. PMID- 17115531 TI - [Oxygen inhalation therapy--low oxygen-flow system with the simple mask]. PMID- 17115532 TI - [Oxygen inhalation therapy-low-oxygen-flow system with OxyArm]. PMID- 17115533 TI - [Low flow oxygen inhalation therapy with the transtracheal catheter system]. PMID- 17115534 TI - [Oxygen inhalation therapy with the high flow oxygen nebuliser]. PMID- 17115535 TI - [High flow oxygen inhalation therapy with the reservoir-attached nasal cannula]. PMID- 17115537 TI - [Not Available]. PMID- 17115536 TI - [Hyperbaric oxygen therapy]. PMID- 17115538 TI - [Not Available]. PMID- 17115539 TI - [Not Available]. PMID- 17115540 TI - [Not Available]. PMID- 17115541 TI - [Not Available]. PMID- 17115543 TI - [Not Available]. PMID- 17115542 TI - [Not Available]. PMID- 17115544 TI - [Not Available]. PMID- 17115545 TI - [Not Available]. PMID- 17115546 TI - [Not Available]. PMID- 17115547 TI - [Not Available]. PMID- 17115548 TI - [Not Available]. PMID- 17115549 TI - [Not Available]. PMID- 17115550 TI - [Not Available]. PMID- 17115551 TI - [Not Available]. PMID- 17115553 TI - [Not Available]. PMID- 17115552 TI - [Not Available]. PMID- 17115554 TI - [Not Available]. PMID- 17115555 TI - [Not Available]. PMID- 17115556 TI - Make CPT codes public. PMID- 17115557 TI - Diagnosing mental illness. PMID- 17115558 TI - Reimbursement shell game. PMID- 17115559 TI - Targeting heart disease. PMID- 17115560 TI - A cost-effectiveness evaluation of single and combined smoking cessation interventions in Texas. PMID- 17115561 TI - Dengue and South Texas: information for clinicians. PMID- 17115562 TI - Texas liability reform an example for the nation. PMID- 17115563 TI - The next exponential leap. PMID- 17115564 TI - Health information technology's potential to improve care: what is the reality? PMID- 17115565 TI - Embracing EMRs: physicians who have done so say change is worth the cost. PMID- 17115566 TI - At your fingertips: EMRs can help comply with pay for performance. PMID- 17115567 TI - Driving the information train: Feds put HIT adoption on fast track. PMID- 17115568 TI - A few tips on avoiding EMR pitfalls. PMID- 17115569 TI - The angel of EMS. PMID- 17115570 TI - Future physicians dissuaded by hassles. PMID- 17115571 TI - TMA takes on insurers for you. PMID- 17115572 TI - Paging Dr. Nurse. PMID- 17115573 TI - Decrease the increase. PMID- 17115574 TI - Pride of ownership. PMID- 17115575 TI - Consent may not matter. PMID- 17115576 TI - Another first for Dr. Burns. PMID- 17115577 TI - What if it happens here? Preparation is the key to surviving catastrophes. PMID- 17115578 TI - A pox on your practice. iPLEDGE program scars dermatologists. PMID- 17115579 TI - Too young. El Paso ordinance curbs underage drinking. PMID- 17115580 TI - Supply versus demand. Number of Texas primary care physicians jumps. PMID- 17115581 TI - Medicine on the front lines. Dr. Rivera returns home after duty in Iraq. PMID- 17115582 TI - Old school. Dr. Homer hopes to improve medicine's professionalism. PMID- 17115583 TI - Investigation of Texas poison center calls regarding a chlorine gas release: implications for terrorist attack toxicosurveillance. AB - The investigation reported here was conducted to describe the pattern of calls received by the Texas Poison Center Network (TPCN) in relation to a chlorine gas release that resulted from a train collision in Bexar County, Texas, on June 28, 2004, and to test various methods for conducting toxicosurveillance. TPCN received a total of 42 calls; the first call was received approximately 35 minutes after the collision. Calls continued for 10 days after the collision. Comparison of the number of calls received from Bexar County on the collision date with the number of similar calls received in the past revealed that numbers for this collision date were elevated for total calls, total information calls, total human exposure calls, chlorine gas calls, and calls involving coughing or choking, headache, throat irritation, or bronchospasm. When a similar analysis was performed for the entire state, call numbers were elevated only for chlorine gas calls and calls involving bronchospasm. PMID- 17115584 TI - Understanding your patients. PMID- 17115585 TI - "Hello, doctor"? PMID- 17115586 TI - Leaving campus. PMID- 17115587 TI - A surgeons's tale: back to West Africa. PMID- 17115588 TI - Hepatitis C virus seroprevalence: selected health care settings in Texas. PMID- 17115589 TI - [New possibilities of surgical treatment of Clatskin tumor and extended focal hepatic affection]. AB - Modern approaches for surgical treatment of hepatic tumors are presented. Results of hepatic resection conduction in 220 patients were analyzed. The indications are substantiated, the main diagnostic methods in hepatic tumors are suggested, and issues on surgical tactics and technique of extended resection performance are put on light. New methods of combined resection of liver and main vessels are elaborated. Their introduction has permitted to lower mortality, to raise the tumors resectability, and to enhance the patients' survival indices. PMID- 17115590 TI - [Modern tactics for choledocholithiasis treatment]. AB - There is presented tactics of treatment of 1325 patients, in whom choledocholithiasis was diagnosed. In 1023 (7.4%) of them in the first stage endoscopic papillosphincterotomy was performed and transpapillar endobiliary manipulations, in 1-2 days--laparoscopic cholecystectomy. In 302 (2.9%) patients laparoscopic exploration of common biliary duct with cholecystectomy were performed. Intraoperative complications occurred in 28 (2.7%) and postoperative- in 72 (7%) patients. All patients are alive. PMID- 17115591 TI - [Preoperative biliary decompression in patients with malignant tumor of duodenal major papilla]. AB - Experience of surgical treatment of 241 patients with malignant tumor of duodenal major papilla in 1992-2005 yrs was analyzed. In 95 patients the treatment was conducted in two stages, the first of which have had included biliary decompression. There were analyzed 18 potential risk factors for postoperative complications occurrence. Preoperative biliary decompression conduction have had constituted one of the risk factors for postoperative complications occurrence. There were proposed strict recommendations concerning preoperative biliary decompression conduction in patients with malignant tumor of duodenal major papilla. PMID- 17115592 TI - [Surgical tactics in the treatment of duodenal ulcer disease coexistent with diseases of other organs of abdominal cavity]. AB - Surgical treatment was performed in 102 patients suffering duodenal ulcer and chronical calculous cholecystitis, gastroesophageal leyomyoma, esophageal, duodenal, small intestinal diverticulum, cardiospasm and other diseases. In all the patients the simultant operative intervention was performed for duodenal ulcer and other diseases of the abdominal cavity organs. All the patients had survived after the operation. Good and fair results were noted in terms of 6 mo 20 yrs follow-up in 79 patients. PMID- 17115593 TI - [Peculiarities of microflora in an acute appendicitis and its impact on the postoperative purulent-inflammatory complications rate]. AB - Bacteriological investigation of processus vermiformis contents was performed in 102 patients suffering an acute appendicitis (AA) and of abdominal cavity exudate -in 82 patients, operated for an AA. The microflora identification was done, as well as probable connection between microorganisms associations, the rate of the AA destructive forms revealing and postoperative purulent-inflammatory complications occurrence. PMID- 17115594 TI - [Application of preperitoneal access for surgical treatment of inguinal hernia]. AB - The results of treatment of 51 patients suffering uncomplicated inguinal hernia were analyzed. There was established that application of preperitoneal access permits to estimate the inguinal channel height and to determine the indications for application of alloplasty. In 29.4% of observations the internal inguinal space height have had exceeded 3.1 cm, predisposing to hernia recurrence in postoperative period. While the internal inguinal space height have constituted 4.1 cm and more it is mandatory to use the net for its secure fixation in the early postoperative period and for prophylaxis of the abdominal wall deformity, setting the net no less than 5 cm behind the muscles edges. In two years after the operation there were examined 24 patients, recurrence was absent. PMID- 17115595 TI - [The splanchnic blood flow indices in the patients, suffering external pancreatic fistula, and their dynamics under the influence of sandostatin and somatulin according to the ultrasonographic duplex scanning data]. AB - Hemodynamical effects of the somatostatin analogues were studied in the patients with external pancreatic fistula. There were examined 29 patients, using ultrasonographic duplex scanning, for investigation of the blood flow indices in a. mesenterica superior, truncus coeliacus and its branches, v. lienalis, v. portae and pancreatic intraorgan arteries. Initial indices of splanchnic blood flow were compared with such while administration of octreotide. The main splanchnic blood flow indices in patients, suffering external pancreatic fistula, did not differ from that in controls, the lowering of the pulsation and resistance indices was noted as well as enhancement of the vessels quantity in pancreatic surgical margin. Under the influence of somatostatin the quantity of visualized vessels had reduced, the pulsation and resistance indices increased, the blood flow velocity along a. mesenterica superior, a. lienalis and pancreatic intraorgan arteries lowered, causing reduction of the blood flow linear and volumetric velocity along v. mesenterica superior and vv. intrapancreatici. Inhibitory action of the preparation on splanchnic blood flow was maximal in terms from 6 till 24 h after its infusion and had lowered step by step during all the follow-up period. PMID- 17115596 TI - [Twenty-years experience of relaparotomy performance]. AB - The results of 204 relaparotomy operations, performed by a single surgeon in 1985 2004 yrs, were analyzed. The data concerning primary operations, their order of emergency, complications, occurred in postoperative period, demanding relaparotomy performance, the causes of their occurrence, were presented. PMID- 17115597 TI - [Pulmonary tissue welding--the method of nonresectional intervention for spontaneous pneumothorax]. AB - New method of the viable tissues welding was applied for spontaneous pneumothorax treatment. There was presented first experience of pulmonary tissue welding without its resection. Since January, 2005 there were performed 17 operations of restricted thoracotomy in randomly selected patients with spontaneous pneumothorax. During the operation there were revealed solitary disseminated bullas in 4 (23.5%) observations, intrapulmonary and superficial--in 8 (47%), thin-wall bullas conglomerate--in 5 (29.5%). In 6 patients up to the operation performance the air passage was preserved. In all the patients the parts of emphysematously changed pulmonary tissue were processed using bipolar set up of original construction. In all the patients there were achieved the bullas electroablation and pulmonary tissue closure without usage of suturing devices or manual suturing. The pneumothorax recurrence, bleeding, durable passage of air, infection complications were not observed. The authors consider the method of emphysematously changed tissue welding a secure one in treatment of spontaneous pneumothorax, omitting pulmonary tissue resection. PMID- 17115598 TI - [Prognostication of the treatment results in patients with angiospasm and subarachnoid hemorrhage, caused by arterial aneurysm]. AB - Treatment of angiospasm as one of the most dangerous complications of subarachnoidal hemorrhage in the cerebral arterial aneurysm rupture constitutes an actual problem. Possibility of the cerebral vessels visualization have permitted to use intraarterial infusion of vasoactive preparations (IAIVAP) for treatment of angiospasm. Therapeutic efficacy for angiospasm, cerebral ischemia and edema depends on many factors and even in application of IAIVAP not always terminates successfully. The disease predictors studying makes possible to plan the intensive therapy tactics correctly, to prognosticate the disease course and the result of treatment. While analyzing the material there were used statistical methods of the data processing, permitting to reveal the prognostically meaningful criterions, influencing the treatment efficacy for angiospasm and its consequences. There was proved the influence of some predictors on the survival and mortality indexes, as well as on disablement and securing of ability to work in patients after application of IAIVAP. It was established, that conduction of IAIVAP had promoted the trustworthy raising of the survival indexes, as well as the lowering of mortality and the disablement rate in patients suffering angiospasm, cerebral ischemia and edema. PMID- 17115599 TI - [The reasons and frequency of early reocclusion origin in vessels of femoro popliteal-tibial segment]. AB - Main reasons and frequency of early postoperative thrombotic complications were analyzed. The frequency of early postoperative thrombosis in vessels of femoro popliteal-tibial segment composed 19.1%. The most frequency of early reocclusion observed after the femoro-tibial reconstructive operations (in 32.4% patients), the least--after femoro-popliteal grafting at above knee level (in 14.1%). The main reason for early reocclusion is an inadequate estimation of distal arteries damage. PMID- 17115600 TI - [Necessity of bladder extirpation performance in the treatment of severe complications of its cancer]. AB - Impact of bladder extirpation on quality of life in the patients, suffering severe complications of bladder cancer, was studied and efficacy of the urine turn off was determined. Results of treatment of 52 patients with complicated course of the bladder cancer, to whom bladderectomy was performed for vital indications using various methods of the urine turn off, were analyzed. After performance of intrarectal turn off of the urine with formation urinal reservoir or ileo-neobladderplasty there was noted the lowest rate of ureterohydronephros occurrence. Comparative analysis on quality of life index in the patients there was revealed its trustful improvement in postoperative period. Overall, positive result of treatment was achieved in more than 90% of patients. PMID- 17115601 TI - [The changes of general potential of hemocoagulation in children, suffering an acute hematogenic osteomyelitis]. AB - Chronometric hypocoagulation was observed in children, suffering an acute hematogenic ostheomyelitis, witnessed by processes of thrombin formation according to internal (the prolonged time of the blood plasm recalcification and activated partial thromboplastin time) and external (the thrombin time enhancement) ways of the blood coagulation process, as well as changes in fibrinogenesis mechanisms (the thrombin time prolongation). The lowering of anticoagulant capacity of the blood (the antithrombin III activity inhibition by 18.5%) was combined with significant increase of the thrombocytes functional activity (the rising of their adhesive and aggregational properties) in more than two times, which have occurred on the background of constant content of fibrinogen in the blood. Changes in the system of the plasm fibrinolysis in an acute hematogenic ostheomyelitis was characterized by inhibition of cofermental and, mainly, fermental fibrinolytic activity of the blood plasm, in conjunction with Hageman-dependent fibrinolysis intensification and was accompanied by accumulation of soluble complexes of fibrin-monomer in the blood. So far, chronometric hypocoagulation is secondary process, caused by the influence of soluble complexes of fibrin-monomer, which blocks fibrinogenesis. That's why the general potential of the blood coagulation system in children with an acute hematogenic ostheomyelitis must be regarded as a structural hypercoagulation. PMID- 17115602 TI - [The influence of photochemically activated 40% solution of levomecol ointment toward apoptosis in muscles of the rats posterior extremities in inflammatory purulent complications of diabetes mellitus]. AB - The influence of photochemically activated 40% solution of Levomecol ointment toward the tissue apoptosis while occurrence of ischemic complications of diabetes mellitus was studied in experiment on 184 adult mongrel white male rats weighted 250-310 g. While modelling of ischemic complications of diabetes mellitus there was revealed the cells apoptosis regulation in soft tissues of the affected lower extremities, causing worsening of the reparative processes course. Application of photochemically activated 40% solution of Levomecol ointment had promoted an effective correction of disorders in the cells apoptosis regulation. It substantiates the expediency of the method introduction into clinical practice. PMID- 17115603 TI - [Perforative gastric ulcer and an acute gastric hemorrhage in a patient with an acute purulent meningoencephalitis]. PMID- 17115604 TI - [Incarcerated posttraumatic diaphragmatic hernia coexistent with duodenal ampulla's ulcer perforation into the left pleural cavity]. PMID- 17115605 TI - [Choice of the method for late reconstruction of the breast for their inborn and acquired defects]. AB - Algorhythm of the method choice in late reconstruction of the breast for postmastectomy syndrome and other defects of mammarial gland was proposed, basing on analysis of own experience of treatment of 57 patients in 1998-2005 yrs. Indications for the breast restoration, using autotissues, endoprostheses, expander-endoprostheses systems and the methods combination are depicted in detail. PMID- 17115606 TI - [Retrospective analysis and modern concept of treatment of the limbs lymphoedema]. AB - Lymphoedema--is a chronic disease, characterized by the volume enhancement of some part of body, more often--of the limbs, as a consequence of tissues oedema, caused by the lymphatic system transport and resorptive function disorder. Modern approaches to lymphoedema treatment foresees combination of conservative and surgical ones, for example, microsurgical, taking into account the oedema character, stage of the disease, the previous treatment efficacy. PMID- 17115607 TI - [Complications of endoscopic transpapillary interventions]. AB - The results of endoscopic papillosphincterotomy with subsequent endobiliary intervention performance in the clinic in 2235 patients in 2000-2006 yrs were studied up. Early complications had occurred in 113 (5.1%) patients, including hemorrhage--in 57 (50%); late complications--in 64 (2.9%). All the patients are alive. PMID- 17115608 TI - [Influence of method of cholecystectomy performance on the central hemodynamics indices in patients with an acute cholecystitis]. AB - There were examined 120 patients, using the method of the integral rheography of the body (according to method of M. I. Tyshchenko), and than operated on for the acute cholecystitis. In 60 patients, constituting the main group, cholecystectomy using minimal approach (CHMA) was done and in 60 (control group)--open cholecystectomy (OCH) using wide laparotomy. After performance of OCH the significant reduction of volumetric indices of blood flow was established. Extremely significant changes of the hemodynamics indices was observed on 1-3 days and than they had slowly improved and on 7-9 day restored, but not completely up to primary data in majority of cases. After performance of CHMA the indices changes were less significant and had restored earlier. PMID- 17115609 TI - [Surgical correction of the biliary ducts injury while performance of laparoscopic cholecystectomy]. AB - There were analyzed the results of treatment of 65 patients, in whom unintended injury of biliary ducts occurred while performance of laparoscopic cholecystectomy in 1993-2005 yrs, including 9 patients, operated in the clinic and 56--in other hospitals. For correction of intraoperatively revealed injuries of biliary ducts there were formated bilio-biliary anastomosis, cholecystoenteroanastomosis--in 1, hepaticojejunoanastomosis end to side (in 2), "high" hepaticojejunoanastomosis (in 5). If the injuries were revealed more lately, application of two-staged treatment was needed in majority of cases. Miniinvasive intervention, as the definitive method of correction, was applied in 11 patients with small injury of biliary ducts, insufficiency of the gallbladder stump sutures due to residual choledocholithiasis, and also as an additive manipulation in the treatment of complete injury of biliary ducts--in 23. Postoperative mortality was 1.5%, and in late period--1.5%. PMID- 17115610 TI - [Optimization of prevention of postoperative purulent-inflammatory complications in patients, operated on for diseases of the abdominal cavity organs]. AB - There were examined 59 patients with diseases of the abdominal cavity organs, in whom high risk of postoperative purulent-inflammatory complications was established. There were conducted 652 different sowing on various stages of operative intervention conduction to study up the operative field and the blood microflora. There was established, that in microbal scenery in the patients of the risk group the mixed bacterial-fungal infection prevails. Complex of prophylactic measures, which approves the reduction of the purulent-inflammatory complications occurrence, was proposed. PMID- 17115611 TI - [Treatment of bilateral inguinal hernia]. AB - Experience of treatment of 130 patients suffering bilateral inguinal hernia was summarized. Intraoperative classification, elaborated by Gilbert-Rutkow and modified for bilateral inguinal hernia, was applied. Collagenic insufficiency signs in fascia transversalis were revealed in patients, suffering bilateral inquinal hernia, according to morphological investigation data obtained. The plasty method, using synthetic prostheses, was choused according to the hernial type established, in 36 patients the Ukrainian polypropylene meshes were implanted. The organism reactivity toward implanted Ukrainian prostheses was similar to that, observed after implantation of foreign analogues. Hernial recurrence was absent; the complications rate was similar to that, observed after implantation of foreign prostheses. The elaborated tactic introduction had promoted the improvement of the surgical treatment results due to postoperative complications rate reduction in 3 times, hernial recurrence--by 10%, the disability period duration--in 1.3 times as well as terms of the patients social rehabilitation. PMID- 17115612 TI - [New pharmacotherapeutic approach to the treatment and prophylaxis of ischemic insult]. AB - Complex analysis of Extra Erbisol influence on cerebral hemodynamics, bioelectric activity of the brain in patients, who survived the ischemic insult, the expediency of the preparation application in complex of treatment and rehabilitation of the patients were suggested. There was established, that Extra Erbisol application had promoted the cerebral hemodynamics improvement and intrahemispheric interrelationship harmonization. This substantiates the recommendations for including of the preparation to the complex treatment of patients, who had survived an ischemic insult. PMID- 17115613 TI - [Estimation of frequency of the main clinical features in patients with malignant tumor of thyroid gland after radical operative intervention]. AB - The rate of main clinical features was studied in 25 patients with the thyroid gland carcinoma after performance of total thyroidectomy, it was compared also with such a rate in patients with stable hypoparathyroidism as a complication of radical operative intervention made on thyroid gland or parathyroid glands. There was established, that nearly all physical features and majority of somatic symptoms are caused by the stable postoperative hypoparathyroidism occurrence. PMID- 17115614 TI - [Application of videothoracoscopy in surgical treatment of the funnel-shaped thorax]. AB - In 2002-2005 yrs in the clinic 3251 patients were operated on, in 42 (1.29%) of them--for the inborn thorax deformity the method of Nuss was applied. There were 40 men and 2 women. Results of treatment were studied in terms from 1 to 3 years, positive effect was noted in all the patients. The complications were not observed. PMID- 17115615 TI - [Principles of patients selection to esthetic plastic surgical clinic]. AB - The original principles of patients selection for esthetic operations performation are presented. Formalized card of registration of medical information that permit to estimate individual peculiarities of any patient is proposed. Presence of functional and organic disturbances in patients show the need of reviewing of present statement that esthetic interventions are performed in healthy patients. PMID- 17115616 TI - [Postvagotomy complications as a cause of unsatisfactory results of organpreserving operations]. PMID- 17115617 TI - [Modern approaches to the breast reconstruction after mastectomy, made for cancer]. PMID- 17115618 TI - [Apparatus for treatment and support]. PMID- 17115619 TI - [Deceleration syndrome in polytrauma]. PMID- 17115620 TI - [Observation of bilateral thoracoabdominal wounding]. PMID- 17115621 TI - [Perforative gastric ulcer in destructive appendicitis]. PMID- 17115622 TI - [Not Available]. PMID- 17115623 TI - [Not Available]. PMID- 17115624 TI - [Not Available]. PMID- 17115625 TI - [Not Available]. PMID- 17115626 TI - [Not Available]. PMID- 17115627 TI - [Not Available]. PMID- 17115628 TI - [Not Available]. PMID- 17115629 TI - [Not Available]. PMID- 17115630 TI - [Not Available]. PMID- 17115631 TI - [Not Available]. PMID- 17115632 TI - [Not Available]. PMID- 17115633 TI - [Not Available]. PMID- 17115634 TI - [Not Available]. PMID- 17115635 TI - [Not Available]. PMID- 17115637 TI - [Not Available]. PMID- 17115636 TI - [Not Available]. PMID- 17115638 TI - [Not Available]. PMID- 17115639 TI - [Not Available]. PMID- 17115641 TI - [Not Available]. PMID- 17115640 TI - [Not Available]. PMID- 17115642 TI - [Not Available]. PMID- 17115643 TI - [Not Available]. PMID- 17115645 TI - [Not Available]. PMID- 17115644 TI - [Not Available]. PMID- 17115646 TI - [Not Available]. PMID- 17115647 TI - [Not Available]. PMID- 17115648 TI - [Not Available]. PMID- 17115649 TI - [Not Available]. PMID- 17115650 TI - [Not Available]. PMID- 17115651 TI - [The world famous professor involved with plagiarism and fraud]. PMID- 17115652 TI - [Discovery of mechanisms behind genetic regulation awarded with Nobel Prize]. PMID- 17115653 TI - [Intraoperative fluid administration. More difficult than believed--methods for customized fluid management available now]. PMID- 17115654 TI - [Molecular medicine tools. Future prospects with new techniques]. PMID- 17115655 TI - [Osteoporosis fractures. A national plan of action required]. PMID- 17115657 TI - [Male osteoporosis--a growing problem]. PMID- 17115656 TI - [Osteoporosis fracture epidemiology]. PMID- 17115658 TI - [Osteoporosis in health economic perspective]. PMID- 17115659 TI - [Fracture panorama in Umea]. PMID- 17115660 TI - [Osteoporosis--a hereditary disease. Physiology and disease mechanisms]. PMID- 17115661 TI - [Bone remodeling]. PMID- 17115662 TI - [Vitamin D deficiency behind osteoporosis]. PMID- 17115663 TI - [Daily exercise strengthens the skeleton]. PMID- 17115664 TI - [Risk fractures of fracture and osteoporosis]. PMID- 17115665 TI - [Osteoporosis screening requires cooperation]. PMID- 17115666 TI - [Measuring methods in research and everyday routines]. PMID- 17115667 TI - [Hip fractures--an enormous public health problem]. PMID- 17115668 TI - [Osteoporosis fractures]. PMID- 17115669 TI - [Falls and hip fractures in elderly can be prevented]. PMID- 17115670 TI - [Special care program for patients with hip fractures]. PMID- 17115671 TI - [Successful cooperation between orthopedic departments and outpatient care]. PMID- 17115672 TI - [What the MONICA project has taught us about cardiovascular diseases--society, public health and health care]. PMID- 17115673 TI - [Treatment of sleep disorders during pregnancy and breast feeding]. PMID- 17115674 TI - [Low carbohydrate diets reduce the need of statin treatment]. PMID- 17115676 TI - [Not Available]. PMID- 17115675 TI - [Psychosomatic disorders--a 150-year retrospective diagnosis of Darwin's symptoms]. PMID- 17115677 TI - [Not Available]. PMID- 17115678 TI - [Not Available]. PMID- 17115679 TI - [Not Available]. PMID- 17115680 TI - [Not Available]. PMID- 17115681 TI - [Not Available]. PMID- 17115682 TI - [Not Available]. PMID- 17115683 TI - [Not Available]. PMID- 17115684 TI - [Not Available]. PMID- 17115685 TI - [Not Available]. PMID- 17115687 TI - [Not Available]. PMID- 17115686 TI - [Not Available]. PMID- 17115688 TI - The Werner and Bloom syndrome proteins catalyze regression of a model replication fork. AB - The premature aging and cancer-prone diseases Werner and Bloom syndromes are caused by loss of function of WRN and BLM proteins, respectively. At the cellular level, WRN or BLM deficiency causes replication abnormalities, DNA damage hypersensitivity, and genome instability, suggesting that these proteins might participate in resolution of replication blockage. Although WRN and BLM are helicases belonging to the RecQ family, both have been recently shown to also facilitate pairing of complementary DNA strands. In this study, we demonstrate that both WRN and BLM (but not other selected helicases) can coordinate their unwinding and pairing activities to regress a model replication fork substrate. Notably, fork regression is widely believed to be the initial step in responding to replication blockage. Our findings suggest that WRN and/or BLM might regress replication forks in vivo as part of a genome maintenance pathway, consistent with the phenotypes of WRN- and BLM-deficient cells. PMID- 17115689 TI - Proteome-wide profiling of isoniazid targets in Mycobacterium tuberculosis. AB - Isoniazid (INH) is an essential drug used to treat tuberculosis. The mycobactericidal agents are INH adducts [INH-NAD(P)] of the pyridine nucleotide coenzymes, which are generated in vivo after INH activation and which bind to, and inhibit, essential enzymes. The NADH-dependent enoyl-ACP reductase (InhA) and the NADPH-dependent dihydrofolate reductase (DfrA) have both been shown to be inhibited by INH-NAD(P) adducts with nanomolar affinity. In this paper, we profiled the Mycobacterium tuberculosis proteome using both the INH-NAD and INH NADP adducts coupled to solid supports and identified, in addition to InhA and DfrA, 16 other proteins that bind these adducts with high affinity. The majority of these are predicted to be pyridine nucleotide-dependent dehydrogenases/reductases. They are involved in many cellular processes, including S-adenosylmethionine-dependent methyl transfer reactions, pyrimidine and valine catabolism, the arginine degradative pathway, proton and potassium transport, stress response, lipid metabolism, and riboflavin biosynthesis. The targeting of multiple enzymes could, thus, account for the pleiotropic effects of, and powerful mycobactericidal properties of, INH. PMID- 17115690 TI - Amphipathic polymers: tools to fold integral membrane proteins to their active form. AB - Among the major obstacles to pharmacological and structural studies of integral membrane proteins (MPs) are their natural scarcity and the difficulty in overproducing them in their native form. MPs can be overexpressed in the non native state as inclusion bodies, but inducing them to achieve their functional three-dimensional structure has proven to be a major challenge. We describe here the use of an amphipathic polymer, amphipol A8-35, as a novel environment that allows both beta-barrel and alpha-helical MPs to fold to their native state, in the absence of detergents or lipids. Amphipols, which are extremely mild surfactants, appear to favor the formation of native intramolecular protein protein interactions over intermolecular or protein-surfactant ones. The feasibility of the approach is demonstrated using as models OmpA and FomA, two outer membrane proteins from the eubacteria Escherichia coli and Fusobacterium nucleatum, respectively, and bacteriorhodopsin, a light-driven proton pump from the plasma membrane of the archaebacterium Halobacterium salinarium. PMID- 17115691 TI - Differences in binding specificity for the homologous gamma- and beta-chain "holes" on fibrinogen: exclusive binding of Ala-His-Arg-Pro-amide by the beta chain hole. AB - The beta-chain amino-terminal sequences of all known mammalian fibrins begin with the sequence Gly-His-Arg-Pro- (GHRP-), but the homologous sequence in chicken fibrin begins with the sequence Ala-His-Arg-Pro- (AHRP-). Nonetheless, chicken fibrinogen binds the synthetic peptide GHRPam, and a previously reported crystal structure has revealed that the binding is in exact conformance with that observed for the human GHRPam-fragment D complex. We now report that human fibrinogen, which is known not to bind APRP, binds the synthetic peptide AHRPam. Moreover, a crystal structure of AHRPam complexed with fragment D from human fibrinogen shows that AHRPam binds exclusively to the beta-chain hole and, unlike GHRPam, not at all to the homologous gamma-chain hole. The difference can be attributed to the methyl group of the alanine residue clashing with a critical carboxyl group in the gammaC hole but being accommodated in the roomier betaC hole where the equivalent carboxyl is situated more flexibly. PMID- 17115692 TI - Structure of the catalytic domain of human protein kinase C beta II complexed with a bisindolylmaleimide inhibitor. AB - The conventional protein kinase C isoform, PKCII, is a signaling kinase activated during the hyperglycemic state and has been associated with the development of microvascular abnormalities associated with diabetes. PKCII, therefore, has been identified as a therapeutic target where inhibitors of its kinase activity are being pursued for treatment of microvascular-related diabetic complications. In this report, we describe the crystal structure of the catalytic domain of PKCbetaII complexed with an inhibitor at 2.6 A resolution. The kinase domain of PKCbetaII was cleaved and purified from full-length PKCbetaII expressed in baculovirus-infected insect cells. The overall kinase domain structure follows the classical bilobal fold and is in its fully activated conformation with three well-defined phosphorylated residues: Thr-500, Thr-641, and Ser-660. Different from the crystal structures of nonconventional PKC isoforms, the C-terminus of the PKCbetaII catalytic domain is almost fully ordered and features a novel alpha helix in the turn motif. An ATP-competitive inhibitor, 2-methyl-1H-indol-3-yl-BIM 1, was crystallized with the PKCbetaII catalytic domain as a dimer of two enzyme inhibitor complexes. The bound inhibitor adopts a nonplanar conformation in the ATP-binding site, with the kinase domain taking on an intermediate, open conformation. This PKCbetaII-inhibitor complex represents the first structural description of any conventional PKC kinase domain. Given the pathogenic role of PKCbetaII in the development of diabetic complications, this structure can serve as a template for the rational design of inhibitors as potential therapeutic agents. PMID- 17115693 TI - Comparison of solution structures and stabilities of native, partially unfolded and partially refolded pepsin. AB - A zymogen-derived protein, pepsin, appears to be incapable of folding to the native state without the presence of the prosegment. To better understand the nature of the irreversible denaturation of pepsin, the present study reports on the characterization of the stability and low-resolution tertiary and secondary structures of native, alkaline unfolded and acid refolded porcine pepsin. Through a combination of small-angle neutron scattering (SANS), CD, and DSC, acid refolded pepsin (Rp) was shown to have secondary and tertiary structures intermediate between the alkaline denatured and native forms but was found to be thermodynamically stable relative to the native state. It was also observed that the acid refolded state of pepsin was dependent on the protein concentration during refolding because CD and SANS data revealed that both the secondary and tertiary structures of concentrated-refolded pepsin (>10 mg/mL) (CRp) were native like, in contrast to the intermediate nature of Rp, refolded under dilute concentration (<10 mg/mL). Despite a native-like conformation, CRp was more stable and had substantially reduced activity compared to that of the native state, suggesting that the protein was misfolded. It is proposed that the stable but misfolded, acid-refolded states are evidence that pepsin in its native conformation was metastable. Furthermore, the disruption of the active site cleft in the denatured states could be discerned by modeling of the SANS data. PMID- 17115694 TI - Electrostatic contributions to residue-specific protonation equilibria and proton binding capacitance for a small protein. AB - Charge-charge interactions in proteins are important in a host of biological processes. Here we use 13C NMR chemical shift data for individual aspartate and glutamate side chain carboxylate groups to accurately detect site-specific protonation equilibria in a variant of the B1 domain of protein G (PGB1-QDD). Carbon chemical shifts are dominated by changes in the electron distribution within the side chain and therefore excellent reporters of the charge state of individual groups, and the data are of high precision. We demonstrate that it is possible to detect local charge interactions within this small protein domain that stretch and skew the chemical shift titration curves away from "ideal" behavior and introduce a framework for the analysis of such convoluted data to study local charge-charge interactions and electrostatic coupling. It is found that, due to changes in electrostatic potential, the proton binding affinity, Ka, of each carboxyl group changes throughout the titration process and results in a linearly pH dependent pKa value. This result could be readily explained by calculations of direct charge-charge interactions based on Coulomb's law. In addition, the slope of pKa versus pH was dependent on screening by salt, and this dependence allowed the selective study of charge-charge interactions. For PGB1 QDD, it was established that mainly differences in self-energy, and not direct charge-charge interactions, are responsible for shifted pKa values within the protein environment. PMID- 17115695 TI - Engineering of a protein with cyclooxygenase and prostacyclin synthase activities that converts arachidonic acid to prostacyclin. AB - Prostacyclin (PGI2), a vascular protector with vasodilation and antithrombotic properties, is synthesized by coupling reactions of cyclooxygenase (COX, the first enzyme) with PGI2 synthase (PGIS, the second enzyme) using arachidonic acid (AA) as an initial substrate. The first COX product, prostaglandin H2 (PGH2) is also a command substrate for other prostanoid enzymes that produce distinct eicosanoids, such as thromboxane A2 (TXA2). The actions of TXA2 to cause vasoconstriction and platelet aggregation oppose the vasodilatory and anti aggregatory effects of PGI2. Specifically upregulating PGI2 biosynthesis is an ideal model for the prevention and treatment of the TXA2-mediated thrombosis involved in strokes and myocardial infarctions. Here, we report that a single protein was constructed by linking COX-2 and PGIS together to form a new fusion enzyme through a transmembrane domain with 10 or 22 residues. The engineered protein expressed in HEK293 and COS-7 cells was able to continually convert AA to prostaglandin (PG) G2 (catalytic step 1), PGH2 (catalytic step 2), and PGI2 (catalytic step 3). The studies first demonstrate that a single protein with three catalytic functions could directly synthesize PGI2 from AA with a Km of approximately 3.2 microM. Specific upregulation of PGI2 biosynthesis through expression of the engineered single protein in the cells has shown strong activity in inhibiting platelet aggregation induced by AA in vitro, which creates a great potential for the fusion enzyme to be used as one of the new therapeutic interventions for strokes and heart attacks. The studies have also provided a model linking COX with its downstream enzymes to specifically regulate biosynthesis of eicosanoids which have potent biological functions. PMID- 17115696 TI - Effects of glycine substitutions on the structure and function of gramicidin a channels. AB - Tryptophan residues often are found at the lipid-aqueous interface region of membrane-spanning proteins, including ion channels, where they are thought to be important determinants of protein structure and function. To better understand how Trp residues modulate the function of membrane-spanning channels, we have examined the effects of Trp replacements on the structure and function of gramicidin A channels. Analogues of gramicidin A in which the Trp residues at positions 9, 11, 13, and 15 were sequentially replaced with Gly were synthesized, and the three-dimensional structure of each analogue was determined using a combination of two-dimensional NMR techniques and distance geometry-simulated annealing structure calculations. Though Trp --> Gly substitutions destabilize the beta6.3-helical gA channel structure, it is possible to determine the structure of analogues with Trp --> Gly substitutions at positions 11, 13, and 15, but not for the analogue with the Trp --> Gly substitution at position 9. The Gly11-, Gly13-, and Gly15-gA analogues form channels that adopt a backbone fold identical to that of native gramicidin A, with only small changes in the side chain conformations of the unsubstituted residues. Single-channel current measurements show that the channel function and lifetime of the analogues are significantly affected by the Trp --> Gly replacements. The conductance variations appear to be caused by sequential removal of the Trp dipoles, which alter the ion-dipole interactions that modulate ion movement. The lifetime variations did not appear to follow a clear pattern. PMID- 17115697 TI - Identification of alpha-1 acid glycoprotein as a lysophospholipid binding protein: a complementary role to albumin in the scavenging of lysophosphatidylcholine. AB - Alpha-1 acid glycoprotein (AGP, orosomucoid), a major acute phase protein in plasma, displays potent cytoprotective and anti-inflammatory activities whose molecular mechanisms are largely unknown. Because AGP binds various exogenous drugs, we have searched for endogenous ligands for AGP. We found that AGP binds lysophospholipids in a manner discernible from albumin in several ways. First, mass spectrometric analyses showed that AGP isolated from plasma and serum contained lysophosphatidylcholine (LPC) enriched in mono and polysaturated acyl chains, whereas albumin contained mostly saturated LPC. Second, AGP bound LPC in a 1:1 molar ratio and with a higher affinity than free fatty acids, whereas albumin bound LPC in a 3:1 ratio but with a lower affinity than that of free fatty acids. Consequently, free fatty acids displaced LPC more avidly from albumin than from AGP. Competitive ligand displacement indicated the highest affinity for AGP to LPC20:4, 18:3, 18:1, and 16:0 (150-180 nM), lysophosphatidylserine (Kd 190 nM), and platelet activating factor (PAF) (Kd 235 nM). The high affinity of AGP to LPC in equilibrium was verified by stopped-flow kinetics, which implicated slow dissociation after fast initial binding, being consistent with an induced-fit mechanism. AGP also bound pyrene-labeled phospholipids directly from vesicles and more efficiently than albumin. AGP prevented LPC-induced priming and PAF-induced activation of human granulocytes, thus indicating scavenging of the cellular effects of the lipid ligands. The results suggest that AGP complements albumin as a lysophospholipid scavenging protein, particularly in inflammatory conditions when the capacity of albumin to sequester LPC becomes impaired. PMID- 17115698 TI - Trapped conformational states of semiquinone (D+*QB-*) formed by B-branch electron transfer at low temperature in Rhodobacter sphaeroides reaction centers. AB - The reaction center (RC) from Rhodobacter sphaeroides captures light energy by electron transfer between quinones QA and QB, involving a conformational gating step. In this work, conformational states of D+*QB-* were trapped (80 K) and studied using EPR spectroscopy in native and mutant RCs that lack QA in which QB was reduced by the bacteriopheophytin along the B-branch. In mutant RCs frozen in the dark, a light induced EPR signal due to D+*QB-* formed in 30% of the sample with low quantum yield (0.2%-20%) and decayed in 6 s. A small signal with similar characteristics was also observed in native RCs. In contrast, the EPR signal due to D+*QB-* in mutant RCs illuminated while freezing formed in approximately 95% of the sample did not decay (tau >107 s) at 80 K (also observed in the native RC). In all samples, the observed g-values were the same (g = 2.0026), indicating that all active QB-*'s were located in a proximal conformation coupled with the nonheme Fe2+. We propose that before electron transfer at 80 K, the majority (approximately 70%) of QB, structurally located in the distal site, was not stably reducible, whereas the minority (approximately 30%) of active configurations was in the proximal site. The large difference in the lifetimes of the unrelaxed and relaxed D+*QB-* states is attributed to the relaxation of protein residues and internal water molecules that stabilize D+*QB-*. These results demonstrate energetically significant conformational changes involved in stabilizing the D+*QB-* state. The unrelaxed and relaxed states can be considered to be the initial and final states along the reaction coordinate for conformationally gated electron transfer. PMID- 17115699 TI - Circular dichroism and magnetic circular dichroism studies of the active site of p53R2 from human and mouse: iron binding and nature of the biferrous site relative to other ribonucleotide reductases. AB - Ribonucleotide reductases (RNR) catalyze the rate-limiting step in the synthesis of deoxyribonucleotides from the corresponding ribonucleotides in the synthesis of DNA. Class I RNR has two subunits: R1 with the substrate binding and active site and R2 with a stable tyrosyl radical and diiron cluster. Biferrous R2 reacts with oxygen to form the tyrosyl radical needed for enzymatic activity. A novel R2 form, p53R2, is a 351-amino acid protein induced by the "tumor suppressor gene" p53. p53R2 has been studied using a combination of circular dichroism, magnetic circular dichroism, variable-temperature variable-field MCD, and EPR spectroscopies. The active site of biferrous p53R2 in both the human (hp53R2) and mouse (mp53R2) forms is found to have one five-coordinate and one four-coordinate iron, which are weakly antiferromagnetically coupled through mu-1,3-carboxylate bridges. These spectroscopic data are very similar to those of Escherichia coli R2, and mouse R2, with a stronger resemblance to data of the former. Titrations of apo-hp53R2 and apo-mp53R2 with Fe(II) were pursued for the purpose of comparing their metal binding affinities to those of other R2s. Both p53R2s were found to have a high affinity for Fe(II), which is different from that of mouse R2 and may reflect differences in the regulation of enzymatic activity, as p53R2 is mainly triggered during DNA repair. The difference in ferrous affinity between mammalian R2 and p53R2 suggests the possibility of specific inhibition of DNA precursor synthesis during cell division. PMID- 17115700 TI - Femtosecond time-resolved absorption spectroscopy of main-form and high-salt peridinin-chlorophyll a-proteins at low temperatures. AB - Steady-state and femtosecond time-resolved optical methods have been used to compare the spectroscopic features and energy transfer dynamics of two systematically different light-harvesting complexes from the dinoflagellate Amphidinium carterae: main-form (MFPCP) and high-salt (HSPCP) peridinin chlorophyll a-proteins. Pigment analysis and X-ray diffraction structure determinations [Hofmann, E., Wrench, P. M., Sharples, F. P., Hiller, R. G., Welte, W., Diederichs, K. (1996) Science 272, 1788-1791; T. Schulte, F. P. Sharples, R. G. Hiller, and E. Hofmann, unpublished results] have revealed the composition and geometric arrangements of the protein-bound chromophores. The MFPCP contains eight peridinins and two chlorophyll (Chl) a, whereas the HSPCP has six peridinins and two Chl a, but both have very similar pigment orientations. Analysis of the absorption spectra has shown that the peridinins and Chls absorb at different wavelengths in the two complexes. Also, in the HSPCP complex, the Qy transitions of the Chls are split into two well-resolved bands. Quantum computations by modified neglect of differential overlap with partial single and double configuration interaction (MNDO-PSDCI) methods have revealed that charged amino acid residues within 8 A of the pigment molecules are responsible for the observed spectral shifts. Femtosecond time-resolved optical spectroscopic kinetic data from both complexes show ultrafast (<130 fs) and slower (approximately 2 ps) pathways for energy transfer from the peridinin excited singlet states to Chl. The Chl-to-Chl energy transfer rate constant for both complexes was measured and is discussed in terms of the Forster mechanism. It was found that, upon direct Chl excitation, the Chl-to-Chl energy transfer rate constant for MFPCP was a factor of 4.2 larger than for HSPCP. It is suggested that this difference arises from a combination of factors including distance between Chls, spectral overlap, and the presence of two additional peridinins in MFPCP that act as polarizable units enhancing the rate of Chl-to Chl energy transfer. The study has revealed specific pigment-protein interactions that control the spectroscopic features and energy transfer dynamics of these light-harvesting complexes. PMID- 17115701 TI - Replacing Asn207 by aspartate at the neck of the D channel in the aa3-type cytochrome c oxidase from Rhodobacter sphaeroides results in decoupling the proton pump. AB - Cytochrome oxidase catalyzes the reduction of O2 to water and conserves the considerable free energy available from this reaction in the form of a proton motive force. For each electron, one proton is electrogenically pumped across the membrane. Of particular interest is the mechanism by which the proton pump operates. Previous studies of the oxidase from Rhodobacter sphaeroides have shown that all of the pumped protons enter the enzyme through the D channel and that a point mutant, N139D, in the D channel completely eliminates proton pumping without reducing oxidase activity. N139 is one of three asparagines near the entrance of the D channel, where there is a narrowing or neck, through which a single file of water molecules pass. In the current work, it is shown that replacement of a second asparagine in this region by an asparate, N207D, also decouples the proton pump without altering the oxidase activity of the enzyme. Previous studies demonstrated that the N139D mutant results in an increase in the apparent pKa of E286, a functionally critical residue that is located 20 A away from N139 at the opposite end of the D channel. In the current work, it is shown that the N207 mutation also increases the apparent pKa of E286. This finding reinforces the proposal that the elimination of proton pumping is the result of an increase of the apparent proton affinity of E286, which, in turn, prevents the timely proton transfer to a proton accepter group within the exit channel of the proton pump. PMID- 17115702 TI - Structural and dynamic repercussions of heme binding and heme-protein cross linking in Synechococcus sp. PCC 7002 hemoglobin. AB - The recombinant two-on-two hemoglobin from the cyanobacterium Synechoccocus sp. PCC 7002 (S7002 rHb) is a bishistidine hexacoordinate globin capable of forming a covalent cross-link between a heme vinyl and a histidine in the C-terminal helix (H helix). Of the two heme axial histidines, His46 (in the E helix, distal side) and His70 (in the F helix, proximal histidine), His46 is displaced by exogenous ligands. S7002 rHb can be readily prepared as an apoglobin (apo-rHb), a non-cross linked hemichrome (ferric iron and histidine axial ligands, rHb-R), and a cross linked hemichrome (rHb-A). To determine the effects of heme binding and subsequent cross-linking, apo-rHb, rHb-R, and rHb-A were subjected to thermal denaturation and 1H/2H exchange. Interpretation of the latter data was based on nuclear magnetic resonance assignments obtained with uniformly 15N- and 13C,15N labeled proteins. Apo-rHb was found to contain a cooperative structural core, which was extended and stabilized by heme binding. Cross-linking resulted in further stabilization attributed mainly to an unfolded-state effect. Protection factors were higher at the cross-link site and near His70 in rHb-A than in rHb-R. In contrast, other regions became less resistant to exchange in rHb-A. These included portions of the B and E helices, which undergo large conformational changes upon exogenous ligand binding. Thus, the cross-link readjusted the dynamic properties of the heme pocket. 1H/2H exchange data also revealed that the B, G, and H helices formed a robust core regardless of the presence of the heme or cross-link. This motif likely encompasses the early folding nucleus of two-on two globins. PMID- 17115703 TI - Structural and functional studies on SCO1815: a beta-ketoacyl-acyl carrier protein reductase from Streptomyces coelicolor A3(2). AB - Aromatic polyketides are medicinally important natural products produced by type II polyketide synthases (PKSs). Some aromatic PKSs are bimodular and include a dedicated initiation module which synthesizes a non-acetate primer unit. Understanding the mechanism of this initiation module is expected to further enhance the potential for regiospecific modification of bacterial aromatic polyketides. A typical initiation module is comprised of a ketosynthase (KS), an acyl carrier protein (ACP), a malonyl-CoA:ACP transacylase (MAT), an acyl-ACP thioesterase, a ketoreductase (KR), a dehydratase (DH), and an enoyl reductase (ER). Thus far, the identities of the ketoreductase, dehydratase, and enoyl reductase remain a mystery because they are not encoded within the PKS biosynthetic gene cluster. Here we report that SCO1815 from Streptomyces coelicolor A3(2), an uncharacterized homologue of a NADPH-dependent ketoreductase, recognizes and reduces the beta-ketoacyl-ACP intermediate from the initiation module of the R1128 PKS. SCO1815 exhibits moderate specificity for both the acyl chain and the thiol donor. The X-ray crystal structure of SCO1815 was determined to 2.0 A. The structure shows that SCO1815 adopts a Rossmann fold and suggests that a conformational change occurs upon cofactor binding. We propose that a positively charged patch formed by three conserved residues is the ACP docking site. Our findings provide new engineering opportunities for incorporating unnatural primer units into novel polyketides and shed light on the biology of yet another cryptic protein in the S. coelicolor genome. PMID- 17115704 TI - Characterization of a Thermobifida fusca beta-1,3-glucanase (Lam81A) with a potential role in plant biomass degradation. AB - Thermobifida fusca is a filamentous soil bacterium that plays a major role in the breakdown of plant biomass. In this paper, we report the cloning, expression, purification, and characterization of the T. fusca enzyme, Lam81A. The Carbohydrate Active Enzymes Database (http://afmb.cnrs-mrs.fr/CAZY/) indicates that Lam81A belongs to a relatively uncharacterized family of beta-1,3 glucanases, family GH-81 [Coutinho, P. M., and Henrissat, B. (1999) in Recent Advances in Carbohydrate Bioengineering (Gilbert, H. J., Davies, G., Henrissat, B., and Svensson, B., Eds.) pp 3-12, The Royal Society of Chemistry, Cambridge, U.K.]. Microarray analysis suggests that Lam81A plays a role in biomass degradation, where its natural substrate may be the plant cell wall polysaccharide, callose, which is a polymer of beta-1,3-linked glucose. Characterization of Lam81A has shown that the enzyme is specific for beta-1,3 linked glucose polysaccharides, is endohydrolytic, and utilizes an inverting mechanism for substrate hydrolysis. In addition, the enzyme has a broad pH optimum from 5.5 to 10, a temperature optimum of 50 degrees C, and demonstrates substrate inhibition, as well as showing a high basal level of expression. PMID- 17115705 TI - A monofunctional and thermostable prephenate dehydratase from the archaeon Methanocaldococcus jannaschii. AB - Prephenate dehydratase (PDT) is an important but poorly characterized enzyme that is involved in the production of L-phenylalanine. Multiple-sequence alignments and a phylogenetic tree suggest that the PDT family has a common structural fold. On the basis of its sequence, the PDT from the extreme thermophile Methanocaldococcus jannaschii (MjPDT) was chosen as a promising representative of this family for pursuing structural and functional studies. The corresponding pheA gene was cloned and expressed in Escherichia coli. It encodes a monofunctional and thermostable enzyme with an N-terminal catalytic domain and a C-terminal regulatory ACT domain. Biophysical characterization suggests a dimeric (62 kDa) protein with mixed alpha/beta secondary structure elements. MjPDT unfolds in a two-state manner (Tm = 94 degrees C), and its free energy of unfolding [DeltaGU(H2O)] is 32.0 kcal/mol. The purified enzyme catalyzes the conversion of prephenate to phenylpyruvate according to Michaelis-Menten kinetics (kcat = 12.3 s-1 and Km = 22 microM at 30 degrees C), and its activity is pH independent over the range of pH 5-10. It is feedback-inhibited by L phenylalanine (Ki = 0.5 microM), but not by L-tyrosine or L-tryptophan. Comparison of its activation parameters (DeltaH(++)= 15 kcal/mol and DeltaS(++)= 3 cal mol-1 K-1) with those for the spontaneous reaction (DeltaH(++) = 17 kcal/mol and DeltaS(++)= -28 cal mol-1 K-1) suggests that MjPDT functions largely as an entropy trap. By providing a highly preorganized microenvironment for the dehydration-decarboxylation sequence, the enzyme may avoid the extensive solvent reorganization that accompanies formation of the carbocationic intermediate in the uncatalyzed reaction. PMID- 17115706 TI - Evolution of multi-enzyme complexes: the case of tryptophan synthase. AB - The prototypical tryptophan synthase is a stable heterotetrameric alpha-betabeta alpha complex. The constituting TrpA and TrpB1 subunits, which are encoded by neighboring genes in the trp operon, activate each other in a bi-directional manner. Recently, a novel class of TrpB2 proteins has been identified, whose members contain additional amino acids that might sterically prevent complex formation with TrpA. To test this hypothesis, we characterized the TrpA and TrpB proteins from Sulfolobus solfataricus. This hyperthermophilic archaeon does not contain a TrpB1 protein but instead contains two TrpB2 homologues that are encoded within (TrpB2i) and outside (TrpB2o) the trp operon. We find that TrpB2i and TrpA form a weak and transient complex during catalysis, with a uni directional activation of TrpA by TrpB2i. In contrast, TrpB2o and TrpA do not form a detectable complex. These results suggest a model for the evolution of the tryptophan synthase in which TrpB2o, TrpB2i, and TrpB1 reflect the stepwise increase of TrpB affinity for TrpA and the refinement of functional subunit interaction, concomitant with the co-localization of the encoding genes in the trp operon. PMID- 17115707 TI - Tryptophan solvent exposure in folded and unfolded states of an SH3 domain by 19F and 1H NMR. AB - The isolated N-terminal SH3 domain of the Drosophila signal transduction protein Drk (drkN SH3) is a useful model for the study of residual structure and fluctuating structure in disordered proteins since it exists in slow exchange between a folded (Fexch) and compact unfolded (Uexch) state in roughly equal proportions under nondenaturing conditions. The single tryptophan residue, Trp36, is believed to play a key role in forming a non-native hydrophobic cluster in the Uexch state, with a number of long-range nuclear Overhauser contacts (NOEs) observed primarily to the indole proton. Substitution of Trp36 for 5-fluoro-Trp36 resulted in a substantial shift in the equilibrium to favor the Fexch state. A variety of 19F NMR measurements were performed to investigate the degree of solvent exposure and hydrophobicity associated with the 5-fluoro position in both the Fexch and Uexch states. Ambient T1 measurements and H2O/D2O solvent isotope effects indicated extensive protein contacts to the 5-fluoro position in the Fexch state and greater solvent exposure in the Uexch state. This was corroborated by the measurements of paramagnetic effects (chemical shift perturbations and T1 relaxation enhancement) from dissolved oxygen at a partial pressure of 20 atm. In contrast, paramagnetic effects from dissolved oxygen revealed less solvent exposure to the indole proton of Trp36 in the Uexch state than that observed for the Fexch state, consistent with the model in which Trp36 indole belongs to a non-native cluster. Thus, although the Uexch state may be described as a dynamically interconverting ensemble of conformers, there appears to be significant asymmetry in the environment of the indole group and the six membered ring or backbone of Trp36. This implied lack of averaging of a side chain position is in contrast to the general view of fluctuating side chains within disordered states. PMID- 17115708 TI - The stoichiometry of host PrPC glycoforms modulates the efficiency of PrPSc formation in vitro. AB - A central event in the formation of infectious prions is the conformational change of a host-encoded glycoprotein, PrPC, into a pathogenic isoform, PrPSc. However, the molecular requirements for efficient PrP conversion remain unknown. In this study, we employed the recently developed protein misfolding cyclic amplification (PMCA) and scrapie cell assay (SCA) techniques to study the role of N-linked glycosylation on prion formation in vitro. The results show that unglycosylated PrPC molecules are required to propagate mouse RML prions, whereas diglycosylated PrPC molecules are required to propagate hamster Sc237 prions. Furthermore, the formation of Sc237 prions is inhibited by substoichiometric levels of hamster unglycosylated PrPC molecules. Thus, interactions between different PrPC glycoforms appear to control the efficiency of prion formation in a species-specific manner. PMID- 17115709 TI - Dimerization and folding processes of Treponema denticola cystalysin: the role of pyridoxal 5'-phosphate. AB - Cystalysin, the key virulence factor in the bacterium Treponema denticola responsible for periodontitis, is a homodimeric pyridoxal 5'-phosphate (PLP)-C-S lyase. The dimerization process and the urea-induced unfolding equilibrium of holocystalysin were compared with those of the apo form. The presence of PLP decreases approximately 4 times the monomer-dimer equilibrium dissociation constant. By using a variety of spectroscopic and analytical procedures, we demonstrated a difference in their unfolding profiles. Upon the monomerization of apocystalysin, occurring between 1 and 2 M urea, a self-associated equilibrium intermediate with a very high beta-sheet content is stabilized over the 2.5-4 M urea range, giving rise to a fully unfolded monomer at higher urea concentrations. On the other hand, highly destabilizing conditions, accompanied by the formation of a significant amount of insoluble aggregates, are required for PLP release and monomerization. Refolding studies, together with analysis of the dissociation/association process of cystalysin, shed light on how the protein concentration and the presence or absence of PLP under refolding conditions could affect the recovery of the active dimeric enzyme and the production of insoluble aggregates. When the protein is completely denatured, the best reactivation yield found was approximately 50% and 25% for holo and apocystalysin, respectively. The dimerization and folding processes of cystalysin have been compared with those of another PLP C-S lyase, MalY from E. coli, and the possible relevance of their PLP binding mode in these processes has been discussed. PMID- 17115710 TI - Interactions of two monoclonal antibodies with BNP: high resolution epitope mapping using fluorescence correlation spectroscopy. AB - Structure-function studies of antibody-antigen systems include the identification of amino acid residues in the antigen that interact with an antibody and elucidation of their individual contributions to binding affinity. We used fluorescence correlation spectroscopy (FCS) and alanine-scanning mutagenesis to characterize the interactions of brain natriuretic peptide (BNP) with two monoclonal antibodies. Human BNP is a 32 amino acid residue long cyclic polypeptide with the ring structure confined between cysteines in positions 10 and 26. It is an important cardiovascular hormone and a valuable diagnostic cardiac marker. We compare the binding strength of the N-terminus Alexa488 labeled BNP, native cyclic BNP, BNP alanine-substituted mutants, linear BNP, and its short fragments to determine the individual contributions of amino acid residues included in the continuous antigenic epitopes that are recognized by two different monoclonal antibodies raised toward BNP. Implementation of FCS for these studies offers all of the advantages of solution phase measurements, including high sensitivity, simplicity of manipulation with reagents, and elimination of solid phase interferences or separation steps. Significant differences in the molecular masses of the free and antibody bound BNP results in a substantial ( approximately 2.5-times) increase in the diffusion rates. Determination of the binding constants and inhibition effects by measuring the diffusion rates of the ligand at the single molecule level introduces the ultimate opportunity for researching systems where the fluorescence intensity and/or fluorescence anisotropy do not change upon interaction of the ligand with the protein. Monoclonal antibodies 106.3 and BC203 demonstrate high affinities to BNP and bind two distant epitopes forming robust antibody sandwiches. Both antibodies are used in Abbott diagnostic assays on AxSYM, IMx, and Architect platforms. PMID- 17115711 TI - Role of the [2Fe-2S]2+ cluster in biotin synthase: mutagenesis of the atypical metal ligand arginine 260. AB - Biotin synthase (BS) is an S-adenosylmethionine (AdoMet)-dependent radical enzyme that catalyzes the addition of sulfur to dethiobiotin. Like other AdoMet radical enzymes, BS contains a [4Fe-4S] cluster that is coordinated by a highly conserved CxxxCxxC sequence motif and by the methionyl amine and carboxylate of AdoMet. The close association of the [4Fe-4S]+ cluster with AdoMet facilitates reductive cleavage of the sulfonium and the generation of transient 5'-deoxyadenosyl radicals, which are then proposed to sequentially abstract hydrogen atoms from the substrate to produce carbon radicals at C9 and C6 of dethiobiotin. BS also contains a [2Fe-2S]2+ cluster located approximately 4-5 A from dethiobiotin, and we have proposed that a bridging sulfide of this cluster quenches the substrate radicals, leading to formation of the thiophane ring of biotin. In BS from Escherichia coli, the [2Fe-2S]2+ cluster is coordinated by cysteines 97, 128, and 188, and the atypical metal ligand, arginine 260. The evolutionary conservation of an arginine guanidinium as a metal ligand suggests a novel role for this residue in tuning the reactivity or stability of the [2Fe-2S]2+ cluster. In this work, we explore the effects of mutagenesis of Arg260 to Ala, Cys, His, and Met. Although perturbations in a number of characteristics of the [2Fe-2S]2+ cluster and the proteins are noted, the reconstituted enzymes have in vitro single turnover activities that are 30-120% of that of the wild type. Further, in vivo expression of each mutant enzyme was sufficient to sustain growth of a bioB- mutant strain on dethiobiotin-supplemented medium, suggesting the enzymes were active and efficiently reconstituted by the in vivo iron-sulfur cluster (ISC) assembly system. Although we cannot exclude an as-yet-unidentified in vivo role in cluster repair or retention, we can conclude that Arg260 is not essential for the catalytic reaction of BS. PMID- 17115712 TI - Deuterium solvent isotope effect and proton-inventory studies of factor Xa catalyzed reactions. AB - Kinetic solvent isotope effects (KSIEs) for the factor Xa (FXa)-catalyzed activation of prothrombin in the presence and absence of factor Va (FVa) and 5.0 x 10(-5) M phospholipid vesicles are slightly inverse, 0.82-0.93, when substrate concentrations are at 0.2 Km. This is consistent with the rate-determining association of the enzyme-prothrombin assembly, rather than the rate-limiting chemical transformation. FVa is known to effect a major conformational change to expose the first scissile bond in prothrombin, which is the likely event triggering a major solvent rearrangement. At prothrombin concentrations > 5 Km, the KSIE is 1.6 +/- 0.3, when FXa is in a 1:1 ratio with FVa but becomes increasingly inverse, 0.30 +/- 0.05 and 0.19 +/- 0.04, when FXa/FVa is 1:4, with an increasing FXa and substrate concentration. The rate-determining step changes with the conditions, but the chemical step is not limiting under any circumstance. This corroborates the proposed predominance of the meizothrombin pathway when FXa is well-saturated with the prothrombin complex. In contrast, the FXa-catalyzed hydrolysis of N-alpha-Z-D-Arg-Gly-Arg-pNA.2HCl (S-2765) and H-D-Ile L-Pro-L-Arg-pNA.HCl (S-2288) is most consistent with two-proton bridges forming at the transition state between Ser195 OgammaH and His57 N(epsilon)2 and His57 Ndelta1 and Asp102 COObeta- at the active site, with transition-state fractionation factors of phi1 = phi2 = 0.57 +/- 0.07 and phiS = 0.78 +/- 0.16 for solvent rearrangement for S-2765 and phi1 = phi2 = 0.674 +/- 0.001 for S-2288 under enzyme saturation with the substrate at pH 8.40 and 25.0 +/- 0.1 degrees C. The rate-determining step(s) in these reactions is most likely the cleavage of the C-N bond and departure of the leaving group. PMID- 17115713 TI - Use of an inhibitor to identify members of the hormone-sensitive lipase family. AB - Hormone-sensitive lipase (HSL) contributes importantly to the mobilization of fatty acids from the triacylglycerols stored in adipocytes, which provide the main source of energy in mammals. On the basis of amino acid sequence alignments and three-dimensional structures, this enzyme was previously found to be a suitable template for defining a family of serine carboxylester hydrolases. In this study, the HSL family members are characterized rather on the basis of their inhibition by 5-methoxy-3-(4-phenoxyphenyl)-3H-[1,3,4]oxadiazol-2-one (compound 7600). This compound inhibits mammalian HSL as well as other HSL family members, such as EST2 from the thermophilic eubacterium Alicyclobacillus acidocaldarius and AFEST from the hyperthermophilic archaeon Archaeoglobus fulgidus. Various carboxylester hydrolases that are not members of the HSL family were found not to be inhibited by compound 7600 under the same experimental conditions. These include nonlipolytic hydrolases such as Torpedo californica acetylcholinesterase and pig liver esterase, as well as lipolytic hydrolases such as human pancreatic lipase, dog gastric lipase, Thermomyces lanuginosus lipase, and Bacillus subtilis LipA. When vinyl esters were used as substrates, the residual activity of HSL, AFEST, and EST2 decreased with an increase in compound 7600 concentration in the incubation mixture. The inhibitor concentration at which the enzyme activity decreased to 50% after incubation for 5 min was 70, 20, and 15 nM with HSL, AFEST, and EST2, respectively. Treating EST2 and AFEST with the inhibitor resulted in an increase in the molecular mass, as established by performing matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis. This increase in the molecular mass, which corresponds approximately to the molecular mass of the inhibitor, indicates that a covalent enzyme-inhibitor complex has been formed. Surface-enhanced laser desorption ionization time-of flight mass spectrometry analysis of a trypsin digest of AFEST treated with the inhibitor or not treated showed the occurrence of an increase in the molecular masses of the "GESAGG"-containing peptide, which is compatible with the formation of a covalent complex with the inhibitor. PMID- 17115715 TI - Hypochlorous acid-derived modification of phospholipids: characterization of aminophospholipids as regulatory molecules for lipid peroxidation. AB - Hypochlorous acid (HOCl), an inflammatory oxidant derived from neutrophil myeloperoxidase, can chlorinate cytosolic proteins and nuclear DNA bases of target cells by passing through the cell membrane. However, little is known about the consequences of HOCl-derived modification of cell membrane components, including phospholipids. In this study, we characterize the reaction of HOCl with phospholipid molecules and found that aminophospholipids are the key molecules that chemically regulate lipid peroxidation. Upon incubation with HOCl, the peroxidation of egg yolk phosphatidylcholine was significantly enhanced in the presence of phosphatidylethanolamine (PE). In contrast, the peroxidation was significantly inhibited in the presence of phosphatidylserine (PS). On the basis of mass spectrometric and electron paramagnetic resonance characterization, the initiator of the peroxidation was identified as the nitrogen-centered radical originating from PE-derived chloramines, especially N,N-dichlorinated PE, a major product in the HOCl-modified PE. Although PS was also chlorinated upon reaction with HOCl, the formed chloramine rapidly decomposed to phosphatidylglycolaldehyde, a novel class of lipid aldehyde. Formation of phosphatidylglycolaldehyde was also confirmed in the porcine brain PS and erythrocyte cell membrane ghost exposed to HOCl. These results provide a novel mechanism for the HOCl-induced oxidative damage and its endogenous protection in the cell membrane at the site of inflammation. PMID- 17115714 TI - Uncoupling of nucleotide flipping and DNA bending by the t4 pyrimidine dimer DNA glycosylase. AB - Bacteriophage T4 pyrimidine dimer glycosylase (T4-Pdg) is a base excision repair protein that incises DNA at cyclobutane pyrimidine dimers that are formed as a consequence of exposure to ultraviolet light. Cocrystallization of T4-Pdg with substrate DNA has shown that the adenosine opposite the 5'-thymine of a thymine thymine (TT) dimer is flipped into an extrahelical conformation and that the DNA backbone is kinked 60 degrees in the enzyme-substrate (ES) complex. To examine the kinetic details of the precatalytic events in the T4-Pdg reaction mechanism, investigations were designed to separately assess nucleotide flipping and DNA bending. The fluorescent adenine base analogue, 2-aminopurine (2-AP), placed opposite an abasic site analogue, tetrahydrofuran, exhibited a 2.8-fold increase in emission intensity when flipped in the ES complex. Using the 2-AP fluorescence signal for nucleotide flipping, kon and koff pre-steady-state kinetic measurements were determined. DNA bending was assessed by fluorescence resonance energy transfer using fluorescent donor-acceptor pairs located at the 5'-ends of oligonucleotides in duplex DNA. The fluorescence intensity of the donor fluorophore was quenched by 15% in the ES complex as a result of an increased efficiency of energy transfer between the labeled ends of the DNA in the bent conformation. Kinetic analyses of the bending signal revealed an off rate that was 2.5-fold faster than the off rate for nucleotide flipping. These results demonstrate that the nucleotide flipping step can be uncoupled from the bending of DNA in the formation of an ES complex. PMID- 17115716 TI - Two toxins from Conus striatus that individually induce tetanic paralysis. AB - We describe structural properties and biological activities of two related O glycosylated peptide toxins isolated from injected (milked) venom of Conus striatus, a piscivorous snail that captures prey by injecting a venom that induces a violent, spastic paralysis. One 30 amino acid toxin is identified as kappaA-SIVA (termed s4a here), and another 37 amino acid toxin, s4b, corresponds to a putative peptide encoded by a previously reported cDNA. We confirm the amino acid sequences and carry out structural analyses of both mature toxins using multiple mass spectrometric techniques. These include electrospray ionization ion trap mass spectrometry and nanoelectrospray techniques for small volume samples, as well as matrix-assisted laser desorption/ionization time of flight mass spectrometric analysis as a complementary method to assist in the determination of posttranslational modifications, including O-linked glycosylation. Physiological experiments indicate that both s4a and s4b induce intense repetitive firing of the frog neuromuscular junction, leading to a tetanic contracture in muscle fiber. These effects apparently involve modification of voltage-gated sodium channels in motor axons. Notably, application of either s4a or s4b alone mimics the biological effects of the whole injected venom on fish prey. PMID- 17115717 TI - Proteins in load-bearing junctions: the histidine-rich metal-binding protein of mussel byssus. AB - Building complex load-bearing scaffolds depends on effective ways of joining functionally different biomacromolecules. The junction between collagen fibers and foamlike adhesive plaques in mussel byssus is robust despite the strikingly dissimilar connected structures. mcfp-4, the matrix protein from this junction, and its presecreted form from the foot tissue of Mytilus californianus were isolated and characterized. mcfp-4 has a mass of approximately 93 kDa as determined by MALDI-TOF mass spectrometry. Its composition is dominated by histidine (22 mol %), but levels of lysine, arginine, and aspartate are also significant. A small amount of 3,4-dihydroxyphenyl-l-alanine (2 mol %) can be detected by amino acid analysis and redox cycling assays. The cDNA-deduced sequence of mcfp-4 reveals multiple variants with highly repetitive internal structures, including approximately 36 tandemly repeated His-rich decapeptides (e.g., HVHTHRVLHK) in the N-terminal half and 16 somewhat more degenerate aspartate-rich undecapeptides (e.g., DDHVNDIAQTA) in the C-terminal half. Incubation of a synthetic peptide based on the His-rich decapeptide with Fe3+, Co2+, Ni2+, Zn2+, and Cu2+ indicates that only Cu is strongly bound. MALDI-TOF mass spectrometry of the peptide modified with diethyl pyrocarbonate before and after Cu binding suggests that histidine residues dominate Cu binding. In contrast, the aspartate-rich undecapeptides preferentially bind Ca2+. mcfp-4 is strategically positioned to function as a macromolecular bifunctional linker by using metal ions to couple its own His-rich domains to the His-rich termini of the preCOLs. Ca2+ may mediate coupling of the C-terminus to other calcium-binding plaque proteins. PMID- 17115719 TI - Christopher Foote's discovery of the role of singlet oxygen [1O2 (1Delta g)] in photosensitized oxidation reactions. AB - The chemistry of singlet molecular oxygen [1O2 (1Delta g)], its importance in atmospheric, biological, and therapeutic processes, and its use as a reagent in organic synthesis have been of considerable interest. Many aspects of singlet oxygen chemistry have emanated from the work of Christopher S. Foote and co workers. Singlet oxygen is a historically interesting molecule with an unusual story connected with its discovery. Foote and Wexler conducted experiments in the 1960s where evidence was obtained supporting 1O2 generation via two independent routes: (1) a photochemical reaction (dye-sensitized photooxidation) and (2) a chemical reaction (NaOCl with H2O2). An important factor in the discovery of 1O2 as the critical reaction intermediate in dye-sensitized photooxygenations was Foote's reassessment of the chemical literature of the 1930s, when 1O2 was suggested to be a viable intermediate in dye-sensitized photooxidation reactions. Experiments that used silica gel beads provided evidence for a volatile diffusible oxidant such as 1O2. However, a contemporaneous quarrel surrounded this early work, and the possible existence of solution-phase 1O2 was ignored for over 2 decades. Not long after Foote's initial studies were published in 1964, the idea of singlet oxygen as an intermediate in photooxidation chemistry gained increasing recognition and verification in organic, gas phase, and biological processes. There are many documented impacts that 1O2 has had and continues to have on biology and medicine, for example, photodynamic therapy and plant defenses. PMID- 17115720 TI - Nucleotide sugar transporters of the Golgi apparatus: from basic science to diseases. AB - Approximately 80% of secreted and membrane proteins (40% of all proteins) of eukaryotes become covalently linked to sugars in the lumen of the Golgi apparatus, a cellular organelle that is part of the secretory system of all eukaryotes. The sugar donors are mostly nucleoside diphosphate sugars (nucleotide sugars) and must be translocated from the cytosol, their site of synthesis, across the Golgi apparatus membrane and into the lumen by specific transporters. These are hydrophobic, homodimeric proteins that span the membrane multiple times. Mutants of these proteins have developmental phenotypes including diseases in humans and cattle. PMID- 17115721 TI - Oxidatively induced abstraction reactions. A synthetic approach to low-coordinate and reactive early transition metal complexes containing metal-ligand multiple bonds. AB - A library of low-coordinate titanium and vanadium complexes containing terminal metal-ligand multiply bonded functionalities such as alkylidenes, alkylidynes, and imides have been prepared by one-electron oxidatively induced alpha-hydrogen abstraction reactions. In the case of the alkylidene motif, the nucleophilic nature of the M-C multiple bond permits subsequent reactions such as alpha hydrogen migration to generate other rare functionalities such as phosphinidene alkyl and imide-alkyls. Identifying and fine-tuning of the supporting ancillary ligand on the metal has allowed the isolation of kinetically stable titanium alkylidene and phosphinidene systems. The former is a key functionality to generate transient titanium alkylidynes, which readily engage in intermolecular C H activation reactions of arenes and alkanes, and the ring-opening metathesis of aromatic substrates such as pyridines. In this Account, we describe several synthetic strategies to achieve reactive functionalities, functionalities that were previously portrayed as "incompatible" or "too kinetically reactive" with 3d early transition metals. PMID- 17115722 TI - Benchmarking state-of-the-art ab initio thermochemical predictions with accurate pulsed-field ionization photoion-photoelectron measurements. AB - This Account presents a comparison between highly precise thermochemical data, including ionization energies, 0 K dissociative photoionization thresholds, and 0 K bond dissociation energies, of selected radicals and molecules and their cations obtained by pulsed field ionization photoion-photoelectron measurements and state-of-the-art thermochemical predictions calculated by the wavefunction based ab initio CCSD(T)/CBS procedures with high-level corrections. The CCSD(T)/CBS method combines the coupled cluster approach including single, double, and quasi-perturbative triple excitations [CCSD(T)] and the complete basis set (CBS) extrapolation approximation. This benchmarking effort indicates that the CCSD(T)/CBS procedures together with high-level corrections are capable of yielding reliable thermochemical predictions with error limits < or =10 meV for small radicals and molecules and their cations. The error limits increase to approximately 35 meV for larger molecular species, such as phenyl and benzyl radicals. PMID- 17115723 TI - Adventures in drug discovery: potent agents based on ligands for cell-surface receptors. AB - How does one go about discovering new drugs? This question is addressed by descriptions of drug discovery research in three project areas that pertain to antagonist ligands for cell-surface receptors. The molecular targets of interest are protease-activated receptor-1 (PAR-1), vasopressin receptors (V1a and V2 subtypes), and the fibrinogen receptor (GPIIb/IIIa). I present different approaches to the identification of high-affinity ligands for these receptors, en route to drug candidates. The PAR-1 project resulted in a pharmacological tool compound that facilitated in vivo proof-of-principle studies, whereas the vasopressin and fibrinogen receptor projects resulted in several preclinical development compounds, three of which advanced into human clinical trials. PMID- 17115724 TI - Synthetic strategies and structural aspects of metal-mediated multiporphyrin assemblies. AB - Porphyrins play a major role as active chromophores in artificial systems mimicking the natural photoinduced processes. The formation of coordination bonds between peripheral donor sites on the porphyrins and external metal fragments has proven to be an efficient alternative to covalent synthesis for the construction of multiporphyrin assemblies, whose complexity and beauty gradually approach those of the multichromophore systems found in nature. In a modular approach, relatively simple metal-mediated porphyrin adducts, owing to their thermodynamic and kinetic stability, can be exploited as building blocks in the construction of higher order architectures. Thus, multichromophore systems become accessible on demand, with a limited synthetic effort. The collection of solid-state structures reported here demonstrates that the flexibility of the porphyrins and the metal junctions, combined with the conformational freedom of the coordination bonds, may lead to assemblies with hardly predictable architectures. Examples in which X ray structural determination was essential for establishing the real composition and geometry of the multiporphyrin assemblies are highlighted. PMID- 17115725 TI - Applications of planar-chiral heterocycles as ligands in asymmetric catalysis. AB - A new family of ligands for asymmetric transition metal-catalyzed reactions has been designed and synthesized. Thus, planar-chiral heterocycles furnish high enantioselectivity in a variety of processes, including isomerizations of allylic alcohols, O-H insertions, and 1,3-dipolar cycloadditions. PMID- 17115726 TI - Excess electrons stabilized on ionic oxide surfaces. AB - Surface excess electrons are remarkable chemical entities that provide great opportunities for the design of new materials with precisely tuned electronic and magnetic properties. In this Account, we describe the structure and electronic properties of excess electron centers generated at the surface of insulating oxides. We also outline the elementary mechanisms that are at the basis of the generation of excess electrons at solid surfaces, setting a comparison to the general problem of excess electron localization in condensed media. Emphasis is given to morphological aspects relative to the surface-trapping sites as deduced from combined electron paramagnetic resonance and accurate quantum chemical calculations. The remarkable reactivity featured by the so formed "electron-rich" surfaces is illustrated, describing the reduction of simple diatomic molecules that form adsorbed radical anions via direct surface to adsorbate electron transfer. PMID- 17115727 TI - Chemistry of metal thio- and selenocarboxylates: precursors for metal sulfide/selenide materials, thin films, and nanocrystals. AB - This Account focuses on recent developments of the chemistry of metal thio- and selenocarboxylates in our laboratory and potential use of some of these compounds as single-source precursors for making metal sulfide and selenide bulk materials, thin films, and nanoparticles. PMID- 17115729 TI - A direct measurement of the dissociation energy of water. AB - We have performed a direct measurement of one of the most fundamental thermochemical values: the O-H bond energy in water. Using a triple-resonance laser excitation scheme, we excite the molecule through a series of vibrational overtone transitions to access directly the onset of the dissociative continuum. The dissociation energy obtained from our experiments, 41145.94+/-0.15 cm(-1), is approximately 30 times more accurate than the currently accepted value and has important implications for other thermochemical quantities linked to the bond energy of water. PMID- 17115730 TI - Resonance energy transfer from a fluorescent dye to a metal nanoparticle. AB - A quantum mechanical theory of the rate of excitation energy transfer from a fluorescent dye molecule to the surface plasmonic modes of a spherical metal nanoparticle is presented. The theory predicts the distance dependence of the transfer rate to vary as 1/d(sigma), with sigma=3-4 at intermediate distances, in partial agreement with the recent experimental results. Forster's 1/d(6) dependence is recovered at large separations. The predicted rate exhibits nontrivial nanoparticle size dependence, ultimately going over to an asymptotic, a(3) size dependence. Unlike in conventional fluorescence resonance energy transfer, the orientational factor is found to vary between 1 and 4. PMID- 17115731 TI - Liquid-liquid transition in supercooled water investigated by interaction with LiCl and Xe. AB - The hypothesis that supercooled water consists of two distinct liquid phases has been explored on the basis of their ability to hydrate nonpolar (Xe) and electrolytic (LiCl) species. Xe incorporated in the bulk of amorphous solid water survives in the deeply supercooled regime above the glass-transition temperature of 136 K and is finally dehydrated at 165 K, whereas LiCl dissolves only in the liquid phase appearing above 165 K. The second liquid phase connects with normal water as inferred from high (poor) solubility of LiCl (Xe). This result also suggests that decoupling of translational diffusion and viscosity in the deeply supercooled regime is caused by domain structures of the two liquid phases formed during a possible liquid-liquid transition. PMID- 17115732 TI - Generalization of the Gaussian electrostatic model: extension to arbitrary angular momentum, distributed multipoles, and speedup with reciprocal space methods. AB - The simulation of biological systems by means of current empirical force fields presents shortcomings due to their lack of accuracy, especially in the description of the nonbonded terms. We have previously introduced a force field based on density fitting termed the Gaussian electrostatic model-0 (GEM-0) J.-P. Piquemal et al. [J. Chem. Phys. 124, 104101 (2006)] that improves the description of the nonbonded interactions. GEM-0 relies on density fitting methodology to reproduce each contribution of the constrained space orbital variation (CSOV) energy decomposition scheme, by expanding the electronic density of the molecule in s-type Gaussian functions centered at specific sites. In the present contribution we extend the Coulomb and exchange components of the force field to auxiliary basis sets of arbitrary angular momentum. Since the basis functions with higher angular momentum have directionality, a reference molecular frame (local frame) formalism is employed for the rotation of the fitted expansion coefficients. In all cases the intermolecular interaction energies are calculated by means of Hermite Gaussian functions using the McMurchie-Davidson [J. Comput. Phys. 26, 218 (1978)] recursion to calculate all the required integrals. Furthermore, the use of Hermite Gaussian functions allows a point multipole decomposition determination at each expansion site. Additionally, the issue of computational speed is investigated by reciprocal space based formalisms which include the particle mesh Ewald (PME) and fast Fourier-Poisson (FFP) methods. Frozen-core (Coulomb and exchange-repulsion) intermolecular interaction results for ten stationary points on the water dimer potential-energy surface, as well as a one-dimensional surface scan for the canonical water dimer, formamide, stacked benzene, and benzene water dimers, are presented. All results show reasonable agreement with the corresponding CSOV calculated reference contributions, around 0.1 and 0.15 kcal/mol error for Coulomb and exchange, respectively. Timing results for single Coulomb energy-force calculations for (H(2)O)(n), n=64, 128, 256, 512, and 1024, in periodic boundary conditions with PME and FFP at two different rms force tolerances are also presented. For the small and intermediate auxiliaries, PME shows faster times than FFP at both accuracies and the advantage of PME widens at higher accuracy, while for the largest auxiliary, the opposite occurs. PMID- 17115733 TI - Hybrid quantum/classical path integral approach for simulation of hydrogen transfer reactions in enzymes. AB - A hybrid quantum/classical path integral Monte Carlo (QC-PIMC) method for calculating the quantum free energy barrier for hydrogen transfer reactions in condensed phases is presented. In this approach, the classical potential of mean force along a collective reaction coordinate is calculated using umbrella sampling techniques in conjunction with molecular dynamics trajectories propagated according to a mapping potential. The quantum contribution is determined for each configuration along the classical trajectory with path integral Monte Carlo calculations in which the beads move according to an effective mapping potential. This type of path integral calculation does not utilize the centroid constraint and can lead to more efficient sampling of the relevant region of conformational space than free-particle path integral sampling. The QC-PIMC method is computationally practical for large systems because the path integral sampling for the quantum nuclei is performed separately from the classical molecular dynamics sampling of the entire system. The utility of the QC-PIMC method is illustrated by an application to hydride transfer in the enzyme dihydrofolate reductase. A comparison of this method to the quantized classical path and grid-based methods for this system is presented. PMID- 17115734 TI - Extended multicanonical method combined with thermodynamically optimized potential: application to the liquid-crystal transition of silicon. AB - A novel method is proposed to study first-order phase transition in real materials. It is applied to the liquid-crystal transition of silicon successfully. It consists of two parts: a direct simulation of the transition by an extended multicanonical ensemble with an order parameter defined with structure factors that characterize the transition, and optimization of a model interatomic potential in terms of the ensemble from an accurate one. These provide a principle to project a first-principles approach on a model-based approach conserving thermodynamic properties of multiple phases. PMID- 17115735 TI - Spherically and system-averaged pair density functional theory. AB - In a couple of recent papers Gori-Giorgi and Savin [Phys. Rev. A 71, 032513 (2005)] proposed a theory that provides simple radial equations to generate the spherically and system averaged pair density. In a recent density matrix functional theory [A. Nagy, Phys. Rev. A 66, 022505 (2002)] it was shown that the problem of an arbitrary system can be reduced to a two-particle problem. Based on this theory, via a double adiabatic connection, it is rigorously derived that the square root of the ground-state spherically and system averaged pair density is the solution of a simple radial equation, that is, contrary to the theory of Gori Giorgi and Savin only a single equation has to be considered. PMID- 17115736 TI - Fully flexible unit cell simulation with recursive thermostat chains. AB - The recursive thermostat chained fully flexible cell molecular dynamic simulation (NsigmaT ensemble) is performed. The ensemble is based on the metric tensor, whose components are used as extended variables. These variables are combined with Nose-Poincare recursive thermostat chains. This extended Hamiltonian approach preserves Hamiltonian in structure, and the partition function satisfies the NsigmaT ensemble state in phase space. In the present study, the generalized leap frog method was employed for time integration. The resulting molecular dynamics simulation was performed for bulk and thin film solid materials in the face-centered-cubic crystal structure. Uniaxial tension test and simple shear test are performed to predict the behaviors of a solid material in the bulk state and nanoscale thin film state. The proposed flexible cell method should serve as a powerful tool for the prediction of mechanical and thermal properties of solid materials including nanoscale behavior. PMID- 17115737 TI - Improved diffusion Monte Carlo propagators for bosonic systems using Ito calculus. AB - The construction of importance sampled diffusion Monte Carlo (DMC) schemes accurate to second order in the time step is discussed. A central aspect in obtaining efficient second order schemes is the numerical solution of the stochastic differential equation (SDE) associated with the Fokker-Plank equation responsible for the importance sampling procedure. In this work, stochastic predictor-corrector schemes solving the SDE and consistent with Ito calculus are used in DMC simulations of helium clusters. These schemes are numerically compared with alternative algorithms obtained by splitting the Fokker-Plank operator, an approach that we analyze using the analytical tools provided by Ito; calculus. The numerical results show that predictor-corrector methods are indeed accurate to second order in the time step and that they present a smaller time step bias and a better efficiency than second order split-operator derived schemes when computing ensemble averages for bosonic systems. The possible extension of the predictor-corrector methods to higher orders is also discussed. PMID- 17115738 TI - Dimer statistics on a Bethe lattice. AB - We discuss the exact solutions of various models of the statistics of dimer coverings of a Bethe lattice. We reproduce the well-known exact result for noninteracting hard-core dimers by both a very simple geometrical argument and a general algebraic formulation for lattice statistical problems. The algebraic formulation enables us to discuss loop corrections for finite dimensional lattices. For the Bethe lattice we also obtain the exact solution when either (a) the dimers interact via a short-range interaction or (b) the underlying lattice is anisotropic. We give the exact solution for a special limit of dimers on a Bethe lattice in a quenched random potential in which we consider the maximal covering of dimers on random clusters at site occupation probability p. Surprisingly the partition function for "maximal coverage" on the Bethe lattice is identical to that for the statistics of branched polymers when the activity for a monomer unit is set equal to -p. Finally we give an exact solution for the number of residual vacancies when hard-core dimers are randomly deposited on a one dimensional lattice. PMID- 17115739 TI - Kohn-Sham perturbation theory: simple solution to variational instability of second order correlation energy functional. AB - The orbital-dependent correlation energy functional resulting from second order Kohn-Sham perturbation theory leads to atomic correlation potentials with correct shell structure and asymptotic behavior. The absolute magnitude of the exact correlation potential, however, is greatly overestimated. In addition, this functional is variationally instable, which shows up for systems with nearly degenerate highest occupied and lowest unoccupied levels like Be. In this contribution we examine the simplest resummation of the Kohn-Sham perturbation series which has the potential to resolve both the inaccuracy and the instability problem of the second order expression. This resummation includes only the hole hole terms of the Epstein-Nesbet series of diagrams, which has the advantage that the resulting functional is computationally as efficient as the pure second order expression. The hole-hole Epstein-Nesbet functional is tested for a number of atoms and ions. It is found to reproduce correlation and ground state energies with an accuracy comparable to that of state-of-the-art generalized gradient approximations. The correlation potential, on the other hand, is dramatically improved compared to that obtained from generalized gradient approximations. The same applies to all quantities directly related to the potential, as, for instance, Kohn-Sham eigenvalues and excitation energies. Most importantly, however, the hole-hole Epstein-Nesbet functional turned out to be variationally stable for all neutral as well as all singly and doubly ionized atoms considered so far, including the case of Be. PMID- 17115740 TI - One-electron properties and electrostatic interaction energies from the expectation value expression and wave function of singles and doubles coupled cluster theory. AB - One-electron density matrices resulting from the explicitly connected commutator expansion of the expectation value were implemented at the singles and doubles coupled cluster (CCSD) level. In the proposed approach the one-electron density matrix is obtained at a little extra cost in comparison to the calculation of the CCSD correlation energy. Therefore, in terms of the computational time the new method is significantly less demanding than the conventional linear-response CCSD theory which requires additionally an expensive calculation of the left-hand solution of the CCSD equations. The quality of the new density matrices was investigated by computing a set of one-electron properties for a series of molecules of varying sizes and comparing the results with data obtained using the full configuration interaction method or higher level coupled cluster theory. It has been found that the results obtained using the new approach are of the same quality as those predicted by the linear-response CCSD method. The novel one electron density matrices have also been applied to study the energy of the electrostatic interaction for a number of van der Waals complexes, including the benzene and azulene dimers. PMID- 17115741 TI - Calculation of smooth potential energy surfaces using local electron correlation methods. AB - The geometry dependence of excitation domains in local correlation methods can lead to noncontinuous potential energy surfaces. We propose a simple domain merging procedure which eliminates this problem in many situations. The method is applied to heterolytic bond dissociations of ketene and propadienone, to SN2 reactions of Cl(-) with alkylchlorides, and in a quantum mechanical/molecular mechanical study of the chorismate mutase enzyme. It is demonstrated that smooth potentials are obtained in all cases. Furthermore, basis set superposition error effects are reduced in local calculations, and it is found that this leads to better basis set convergence when computing barrier heights or weak interactions. When the electronic structure strongly changes between reactants or products and the transition state, the domain merging procedure leads to a balanced description of all structures and accurate barrier heights. PMID- 17115742 TI - Long-range excitations in time-dependent density functional theory. AB - Adiabatic time-dependent density functional theory fails for excitations of a heteroatomic molecule composed of two open-shell fragments at large separation. Strong frequency dependence of the exchange-correlation kernel is necessary for both local and charge-transfer excitations. The root of this is the static correlation created by the step in the exact Kohn-Sham ground-state potential between the two fragments. An approximate nonempirical kernel is derived for excited molecular dissociation curves at large separation. Our result is also relevant when the usual local and semilocal approximations are used for the ground-state potential, as static correlation there arises from the coalescence of the highest occupied and lowest unoccupied orbital energies as the molecule dissociates. PMID- 17115743 TI - The nature of the hydrogen bond: a synthesis from the interacting quantum atoms picture. AB - The interacting quantum atoms approach [IQA, as presented by Blanco et al., J. Chem. Theory Comput. 1, 1096 (2005)] is applied to standard hydrogen bonded dimers. IQA is an interpretation tool based on a real space energy decomposition scheme fully consistent with the quantum theory of atoms in molecules. It provides a partition of every physical term present in the Hamiltonian into atomic and interatomic contributions. The procedure is orbital-free and self contained, needing neither external references nor artificial intermediate states. Binding is the result of a competition between the destabilizing deformations suffered by the interacting fragments upon interaction and the stabilizing interaction energy itself. According to IQA, there is no incompatibility between the prevalent electrostatic image of hydrogen bonded systems and that favoring important covalent contributions. Depending on how we gather the different energetic terms, we may recover electrostatic or covalent pictures from the same underlying quantum mechanical description. Our results show that the nonclassical contributions to hydrogen bonding are spatially localized, involving only the H atom and its two nearest neighbors. IQA is well suited as a comparative tool. Its thin energetic decomposition allows us to recover exactly (or to a very good approximation) the quantities of the most widely used energy decomposition schemes. Such a comparison sheds light on the virtues and faults of the different methods and on the origin of the 50 years old debate regarding the covalent/electrostatic nature of the hydrogen bond. PMID- 17115744 TI - Relativistic heavy-atom effects on heavy-atom nuclear shieldings. AB - The principal relativistic heavy-atom effects on the nuclear magnetic resonance (NMR) shielding tensor of the heavy atom itself (HAHA effects) are calculated using ab initio methods at the level of the Breit-Pauli Hamiltonian. This is the first systematic study of the main HAHA effects on nuclear shielding and chemical shift by perturbational relativistic approach. The dependence of the HAHA effects on the chemical environment of the heavy atom is investigated for the closed shell X(2+), X(4+), XH(2), and XH(3) (-) (X=Si-Pb) as well as X(3+), XH(3), and XF(3) (X=P-Bi) systems. Fully relativistic Dirac-Hartree-Fock calculations are carried out for comparison. It is necessary in the Breit-Pauli approach to include the second-order magnetic-field-dependent spin-orbit (SO) shielding contribution as it is the larger SO term in XH(3) (-), XH(3), and XF(3), and is equally large in XH(2) as the conventional, third-order field-independent spin orbit contribution. Considering the chemical shift, the third-order SO mechanism contributes two-thirds of the difference of approximately 1500 ppm between BiH(3) and BiF(3). The second-order SO mechanism and the numerically largest relativistic effect, which arises from the cross-term contribution of the Fermi contact hyperfine interaction and the relativistically modified spin-Zeeman interaction (FC/SZ-KE), are isotropic and practically independent of electron correlation effects as well as the chemical environment of the heavy atom. The third-order SO terms depend on these factors and contribute both to heavy-atom shielding anisotropy and NMR chemical shifts. While a qualitative picture of heavy-atom chemical shifts is already obtained at the nonrelativistic level of theory, reliable shifts may be expected after including the third-order SO contributions only, especially when calculations are carried out at correlated level. The FC/SZ-KE contribution to shielding is almost completely produced in the s orbitals of the heavy atom, with values diminishing with the principal quantum number. The relative contributions converge to universal fractions for the core and subvalence ns shells. The valence shell contribution is negligible, which explains the HAHA characteristics of the FC/SZ-KE term. Although the nonrelativistic theory gives correct chemical shift trends in present systems, the third-order SO-I terms are necessary for more reliable predictions. All of the presently considered relativistic corrections provide significant HAHA contributions to absolute shielding in heavy atoms. PMID- 17115745 TI - Comparison of free energy methods for molecular systems. AB - We present a detailed comparison of computational efficiency and precision for several free energy difference (DeltaF) methods. The analysis includes both equilibrium and nonequilibrium approaches, and distinguishes between unidirectional and bidirectional methodologies. We are primarily interested in comparing two recently proposed approaches, adaptive integration, and single ensemble path sampling to more established methodologies. As test cases, we study relative solvation free energies of large changes to the size or charge of a Lennard-Jones particle in explicit water. The results show that, for the systems used in this study, both adaptive integration and path sampling offer unique advantages over the more traditional approaches. Specifically, adaptive integration is found to provide very precise long-simulation DeltaF estimates as compared to other methods used in this report, while also offering rapid estimation of DeltaF. The results demonstrate that the adaptive integration approach is the best overall method for the systems studied here. The single ensemble path sampling approach is found to be superior to ordinary Jarzynski averaging for the unidirectional, "fast-growth" nonequilibrium case. Closer examination of the path sampling approach on a two-dimensional system suggests it may be the overall method of choice when conformational sampling barriers are high. However, it appears that the free energy landscapes for the systems used in this study have rather modest configurational sampling barriers. PMID- 17115746 TI - Mass-dependent and non-mass-dependent isotope effects in ozone photolysis: resolving theory and experiments. AB - In addition to the anomalous (17)O and (18)O isotope effects in the three-body ozone formation reaction O+O(2)+M, isotope effects in the destruction of ozone by photolysis may also play a role in determining the isotopic composition of ozone and other trace gases in the atmosphere. While previous experiments on ozone photolysis at 254 nm were interpreted as evidence for preferential loss of light ozone that is anomalous (or "non-mass-dependent"), recent semiempirical theoretical calculations predicted a preferential loss of heavy ozone at that wavelength that is mass dependent. Through photochemical modeling results presented here, we resolve this apparent contradiction between experiment and theory. Specifically, we show that the formation of ozone during the UV photolysis experiments is not negligible, as had been assumed, and that the well known non-mass-dependent isotope effects in ozone formation can account for the non-mass-dependent enrichment of the heavy isotopologs of ozone observed in the experiment. Thus, no unusual non-mass-dependent fractionation in ozone photolysis must be invoked to explain the experimental results. Furthermore, we show that theoretical predictions of a mass-dependent preferential loss of the heavy isotopologs of ozone during UV photolysis are not inconsistent with the experimental data, particularly if mass-dependent isotope effects in the chemical loss reactions of ozone during the photolysis experiments or experimental artifacts enrich the remaining ozone in (17)O and (18)O. Before the calculated fractionation factors can be quantitatively evaluated, however, further investigation of possible mass-dependent isotope effects in the reactions of ozone with O((1)D), O((3)P), O(2)((1)Delta), and O(2)((1)Sigma) is needed through experiments we suggest here. PMID- 17115747 TI - High resolution photofragment translational spectroscopy studies of the near ultraviolet photolysis of imidazole. AB - The fragmentation dynamics of imidazole molecules following excitation at 193.3 nm and at many wavelengths in the range of 210< or =lambda(phot)< or =240 nm have been investigated by H Rydberg atom photofragment translational spectroscopy. Long wavelength excitation within this range results in population of the 1 (1)A(")((1)pisigma(*)) excited state, but the 2 (1)A(')<--X (1)A(')(pi(*)<--pi) transition becomes the dominant absorption once lambda(phot)< or =220 nm. The measured energy disposals show parallels with those found in recent studies of the UV photolysis of pyrrole [Cronin et al., Phys Chem. Chem. Phys. 6, 5031 (2004)]. The total kinetic energy release (TKER) spectra display a "fast" feature, centred at TKER approximately 9200 cm(-1). The analysis of the structure evident in the fast feature reveals the selective population of specific in-plane stretching vibrational levels of the imidazolyl cofragment; these fragments are deduced to carry only modest amounts of rotational excitation. Comparison with calculated normal mode vibrational frequencies allows the assignment of the populated levels and a precise determination of the N-H bond strength in imidazole: D(0)=33,240+/-40 cm(-1). The observed energy disposal can be rationalized using Franck-Condon arguments, assuming that the potential energy surface (PES) for the 1 (1)A(")((1)pisigma(*)) state has a topology similar to that of the corresponding (1)pisigma(*) state of pyrrole. As in pyrrole, photoexcitation populates skeletal motions in the S(1) state (in-plane motions in the present case) that are only weakly coupled to the N-H dissociation coordinate and thus map through into the corresponding product vibrations. A second, "slow" feature is increasingly evident in TKER spectra recorded at shorter lambda(phot). This component, which exhibits no recoil anisotropy, is attributed to H atoms formed by the "statistical" decay of highly vibrationally excited ground state molecules. The form of the TKER spectra observed at short lambda(phot) is rationalized by assuming two possible decay routes for imidazole molecules excited to the 2 (1)A(')((1)pipi(*)) state. One involves fast 2 (1)A(')((1)pipi(*)) right arrow-wavy 1 (1)A(")((1)pisigma(*)) radiationless transfer and subsequent fragmentation on the 1 (1)A(')((1)pisigma(*)) PES, yielding fast H atoms (and imidazolyl cofragments)-reminiscent of behavior seen at longer excitation wavelengths where the 1 (1)A(")((1)pisigma(*)) PES is accessed directly. The second is assumed to involve radiationless transfer to the ground state, most probably by successive 2 (1)A(') right arrow-wavy 1 (1)A(") right arrow-wavy X (1)A(') couplings, mediated by conical intersections between the relevant PESs and the subsequent unimolecular decay of the resulting highly vibrationally excited ground state molecules yielding slow H atoms. PMID- 17115748 TI - Two size regimes of methanol clusters produced by adiabatic expansion. AB - Free neutral methanol clusters produced by adiabatic expansion have been studied by photoelectron spectroscopy and line shape modeling. The results show that clusters belonging to two distinct size regimes can be produced by changing the expansion conditions. While the larger size regime can be well described by line shapes calculated for clusters consisting of hundreds of molecules, the smaller size regime corresponds to methanol oligomers, predominantly of cyclic structure. There is little contribution from dimers to the spectra. PMID- 17115749 TI - Classical line shapes based on analytical solutions of bimolecular trajectories in collision induced emission. AB - The classical theory of collision induced emission (CIE) from pairs of dissimilar atoms is studied. This radiation emitted by accelerating dipoles is produced essentially by their overlap and exchange interaction at short range. This classical calculation is based on exact collinear solutions of the trajectories of two spherical particles in a head-on collision under the influence of a Morse potential with a well of arbitrary depth. Without too restrictive conditions a variety of simple solutions in closed form is obtained that illustrates the connection between collision dynamics and CIE. Depending on the well depth and energy of the particles, they may radiate in the far infrared to infrared or visible and shorter wavelength regions. The results are characterized by a fixed total energy. Nevertheless, a connection is demonstrated between the line shape in CIE computed through the trajectory and that based on the emission of a thermally equilibrated system. PMID- 17115750 TI - Combined nonadiabatic transition-state theory and ab initio molecular dynamics study on selectivity of the alpha and beta bond fissions in photodissociation of bromoacetyl chloride. AB - The selectivity of the alpha C-Cl and beta C-Br bond fissions upon n-->pi(*) excitation of bromoacetyl chloride has been investigated with combined nonadiabatic Rice-Ramsperger-Kassel-Marcus theory and ab initio molecular dynamics calculations, which are based on the potential energy profiles calculated with the complete active space self-consistent field and multireference configuration interaction methods. The Zhu-Nakamura [J. Chem. Phys. 101, 10630 (1994); 102, 7448 (1995)] theory is chosen to calculate the nonadiabatic hopping probability. It is found that nonadiabatic effect plays an important role in determining selective dissociations of the C-Cl and C-Br bonds. The calculated rate constants are close to those from experimentally inferred values, but the branching ratio of the alpha C-Cl and beta C-Br bond fissions is different from the experimental findings. The direct molecular dynamics calculations predict that fission of the C-Cl bond occurs on a time scale of picoseconds and cleavage of the beta C-Br bond proceeds with less probability within the same period. This reveals that the initial relaxation dynamics is probably another important factor that influences the selectivity of the C-Cl and C-Br bond fissions in photodissociation of BrCH(2)COCl at 248 nm. PMID- 17115751 TI - Theoretical investigation of the stability of highly charged C60 molecules produced with intense near-infrared laser pulses. AB - We theoretically investigated the stability of highly charged C(60) (z+) cations produced from C(60) with an ultrashort intense laser pulse of lambda approximately 1800 nm. We first calculated the equilibrium structures and vibrational frequencies of C(60) (z+) as well as C(60). We then calculated key energies relevant to dissociation of C(60) (z+), such as the excess vibrational energy acquired upon sudden tunnel ionization from C(60). By comparing the magnitudes of the calculated energies, we found that C(60) (z+) cations up to z approximately 12 can be produced as a stable or quasistable (microsecond-order lifetime) intact parent cation, in agreement with the recent experimental report by V. R. Bhardwaj et al. [Phys. Rev. Lett. 93, 043001 (2004)] that almost only intact parent C(60) (z+) cations up to z=12 are detected by a mass spectrometer. The results of Rice-Ramsperger-Kassel-Marcus calculation suggest that the lifetime of C(60) (z+) drastically decreases by ten orders of magnitude as z increases from z=11 to z=13. Using the time-dependent adiabatic state approach, we also investigated the vibrational excitation of C(60) and C(60) (z+) by an intense near-infrared pulse. The results indicate that large-amplitude vibration with energy of >10 eV is induced in the delocalized h(g)(1)-like mode of C(60) (z+). PMID- 17115752 TI - Br(2Pj) atom formation dynamics in ultraviolet photodissociation of tert-butyl bromide and iso-butyl bromide. AB - The photodissociation dynamics of tert-C(4)H(9)Br and iso-C(4)H(9)Br has been studied at 234 and 265 nm using two-dimensional velocity map imaging technique. The translational energy and angular distributions have been analyzed for Br, Br(*), and tert-C(4)H(9) radical. The energy distribution of Br atom in the photodissociation of tert-C(4)H(9)Br is found to consist of two Gaussian components. The two components are correlated to two independent reaction paths on the excited potential energy surfaces: (1) the high-energy component from the prompt dissociation along the C-Br stretching mode and (2) the low-energy component from the repulsive mode along the C-Br stretching, coupled with some bending motions. For the energy distribution of Br(*) atom in the photodissociation of tert-C(4)H(9)Br, a third multiphoton dissociative ionization channel is observed at 265 nm in addition to the two energy components corresponding to channels (1) and (2). The energy distributions of Br and Br(*) atoms in the photodissociation of iso-C(4)H(9)Br can be fitted using only one Gaussian function indicating a single formation channel. Relative quantum yields for Br((2)P(32)) at 234 and 265 nm in the photodissociation of tert-C(4)H(9)Br are measured to be 0.76 and 0.65, respectively. For iso-C(4)H(9)Br, the measured value is Phi(234 nm)(Br)=0.81. The contribution of bending modes to Br and Br(*) is much more obvious in the photodissociation of tert-C(4)H(9)Br than in iso C(4)H(9)Br. PMID- 17115753 TI - IR plus vacuum ultraviolet spectroscopy of neutral and ionic organic acid molecules and clusters: acetic acid. AB - Infrared (IR) vibrational spectroscopy of acetic acid (A) neutral and ionic monomers and clusters, employing vacuum ultraviolet (VUV), 10.5 eV single photon ionization of supersonically expanded and cooled acetic acid samples, is presented and discussed. Molecular and cluster species are identified by time of flight mass spectroscopy: the major mass features observed are A(n)H(+) (n=1-9), ACOOH(+) (VUV ionization) without IR radiation present, and A(+) with both IR and VUV radiation present. The intense feature ACOOH(+) arises from the cleavage of (A)(2) at the beta-CC bond to generate ACOOH(+)+CH(3) following ionization. The vibrational spectrum of monomeric acetic acid (2500-7500 cm(-1)) is measured by nonresonant ionization detected infrared (NRID-IR) spectroscopy. The fundamentals and overtones of the CH and OH stretches and some combination bands are identified in the spectrum. Mass selected IR spectra of neutral and cationic acetic acid clusters are measured in the 2500-3800 cm(-1) range employing nonresonant ionization dip-IR and IR photodissociation (IRPD) spectroscopies, respectively. Characteristic bands observed at approximately 2500-2900 cm(-1) for the cyclic ring dimer are identified and tentatively assigned. For large neutral acetic acid clusters A(n)(n>2), spectra display only hydrogen bonded OH stretch features, while the CH modes (2500-2900 cm(-1)) do not change with cluster size n. The IRPD spectra of protonated (cationic) acetic acid clusters A(n)H(+) (n=1 7) exhibit a blueshift of the free OH stretch with increasing n. These bands finally disappear for n> or =6, and one broad and weak band due to hydrogen bonded OH stretch vibrations at approximately 3350 cm(-1) is detected. These results indicate that at least one OH group is not involved in the hydrogen bonding network for the smaller (n< or =5) A(n)H(+) species. The disappearance of the free OH stretch feature at n> or =6 suggests that closed cyclic structures form for A(n)H(+) for the larger clusters (n> or =6). PMID- 17115754 TI - IR plus vacuum ultraviolet spectroscopy of neutral and ionic organic acid monomers and clusters: propanoic acid. AB - The vibrational spectrum of molecular propanoic acid, cooled in a supersonic expansion, in the region of 2500 to 7500 cm(-1) is obtained employing infrared plus vacuum ultraviolet nonresonant ionization detected spectroscopy. The fundamental and first overtone of the CH and OH stretch modes of cold propanoic acid molecules can be identified in the spectrum. Propanoic acid neutral and ionic clusters are also studied employing nonresonant ion dip and photodissociation spectroscopic techniques, respectively. For the neutral dimer, a sequence of features observed at ca. 2500-2700 cm(-1) can be assigned as combination bands of low frequency modes with the COH bending overtone; these features characterize the cyclic dimer ring structure. IR spectra of the larger neutral clusters n=3, 4, 5 indicate that they also have cyclic structures in which the OH groups are engaged in the cluster hydrogen bonding network. The CH groups are not involved in this hydrogen bonding structure. Free OH features are observed for the protonated ion clusters (C(2)H(5)COOH)(n)H(+), n=1,...,5, indicating that at least one OH group of these cluster ions is not involved in the cluster hydrogen bonding network. A comparison of the results for four hydrogen bonding neutral and ionic clusters (CH(3)OH, C(2)H(5)OH, CH(3)COOH, and C(2)H(5)COOH) is presented and discussed. PMID- 17115755 TI - Photodissociation spectroscopy of the Mg+-acetic acid complex. AB - We have studied the structure and photodissociation of Mg(+)-acetic acid clusters. Ab initio calculations suggest four relatively strongly bound ground state isomers for the [MgC(2)H(4)O(2)](+) complex. These isomers include the cis and trans forms of the Mg(+)-acetic acid association complex with Mg(+) bonded to the carbonyl O atom of acetic acid, the Mg(+)-acetic acid association complex with Mg(+) bonded to the hydroxyl O atom of acetic acid, or to a Mg(+)-ethenediol association complex. Photodissociation through the Mg(+)-based 3p<--3s absorption bands in the near UV leads to direct (nonreactive) and reactive dissociation products: Mg(+), MgOH(+), Mg(H(2)O)(+), CH(3)CO(+), and MgCH(3) (+). At low energies the dominant reactive quenching pathway is through dehydration to Mg(H(2)O)(+), but additional reaction channels involving C-H and C-C bond activation are also open at higher energies. PMID- 17115756 TI - Dissociative ionization of ethanol by 400 nm femtosecond laser pulses. AB - The dissociative ionization of ethanol in short-pulsed laser fields at approximately 400 nm is investigated. The yield ratio of the C-O bond breaking with respect to the C-C bond breaking increases sharply as the temporal width increases from 60 to 400 fs, and the yield ratio is two to three times as large as that at 800 nm in the entire pulse-width range of 60-580 fs. The enhancement of the C-O bond breaking of singly charged ethanol at 400 nm and the bond elongation prior to the Coulomb explosion of doubly charged ethanol occurring in the relatively weak light field intensity of 10(12)-10(13) W cm(2) is interpreted by the efficient light-induced coupling among the electronic states at the shorter wavelength of 400 nm. From the double pulse experiment, in which ethanol is irradiated with a pair of short pulses (<80 fs), the most efficient coupling occurs at Deltat=160 fs that is much earlier than Deltat=250 at 800 nm, where Deltat denotes the temporal separation of the two pulses, indicating that the nonadiabatic field-induced potential crossings of singly charged ethanol occurs much earlier at 400 nm than at 800 nm. PMID- 17115757 TI - Ab initio potential energy surfaces, bound states, and electronic spectrum of the Ar-SH complex. AB - New ab initio potential energy surfaces for the (2)Pi ground electronic state of the Ar-SH complex are presented, calculated at the RCCSD(T)/aug-cc-pV5Z level. Weakly bound rotation-vibration levels are calculated using coupled-channel methods that properly account for the coupling between the two electronic states. The resulting wave functions are analyzed and a new adiabatic approximation including spin-orbit coupling is proposed. The ground-state wave functions are combined with those obtained for the excited (2)Sigma(+) state [D. M. Hirst, R. J. Doyle, and S. R. Mackenzie, Phys. Chem. Chem. Phys. 6, 5463 (2004)] to produce transition dipole moments. Modeling the transition intensities as a combination of these dipole moments and calculated lifetime values [A. B. McCoy, J. Chem. Phys. 109, 170 (1998)] leads to a good representation of the experimental fluorescence excitation spectrum [M.-C. Yang, A. P. Salzberg, B.-C. Chang, C. C. Carter, and T. A. Miller, J. Chem. Phys. 98, 4301 (1993)]. PMID- 17115758 TI - Solvent configuration-interaction calculations of intermolecular states in molecule-(atom)N clusters: application to Br2-4HeN. AB - We describe variational calculations of J=0 intermolecular states in Br(2) (4)He(N) clusters. The method employed is analogous to configuration-interaction calculations in electronic-structure work and relies on the ability to express the intermolecular Hamiltonian H(v) as a sum of one- and two-body terms. A basis set is built up from solutions to the Schrodinger equation in which only the one body terms of H(v) are included. These configurations are products of N=1 eigenstates. The matrix of H(v) in a symmetry-adapted configuration basis is then computed, the two-body terms of H(v) serving to couple different configurations. This computation involves integrals of dimension five or less. Filter diagonalization is then used to obtain energies and eigenfunctions within a selected energy range. Results on clusters having N=2-5 are reported. PMID- 17115759 TI - Two-photon absorption cross sections: an investigation of solvent effects. Theoretical studies on formaldehyde and water. AB - The effects of a solvent on the two-photon absorption of microsolvated formaldehyde and liquid water have been studied using hybrid coupled cluster/molecular mechanics (CC/MM) response theory. Both water and formaldehyde were considered solvated in water, where the solvent water molecules were described within the framework of molecular mechanics. Prior to the CC/MM calculations, molecular dynamics simulations were performed on the water/formaldehyde and water/water aggregates and many configurations were generated. By carrying out CC/MM response calculations on the individual configurations, it was possible to obtain statistically averaged results for both the excitation energies and two-photon absorption cross sections. For liquid water, the comparison between one- and two-photon absorption spectra is in good agreement with the experimental data available in the literature. In particular, the lowest energy transition occurring in the one-photon absorption spectrum of water only occurs with a relatively small strength in the two-photon absorption spectrum. This result is important for the interpretation of two-photon absorption data as these results show that in the absence of selection rules that determine which transitions are forbidden, the spectral profile of the two-photon absorption spectrum can be significantly different from the spectral profile of the one-photon absorption spectrum. PMID- 17115760 TI - Effective Hamiltonians by optimal control: solid-state NMR double-quantum planar and isotropic dipolar recoupling. AB - We report the use of optimal control algorithms for tailoring the effective Hamiltonians in nuclear magnetic resonance (NMR) spectroscopy through sophisticated radio-frequency (rf) pulse irradiation. Specifically, we address dipolar recoupling in solid-state NMR of powder samples for which case pulse sequences offering evolution under planar double-quantum and isotropic mixing dipolar coupling Hamiltonians are designed. The pulse sequences are constructed numerically to cope with a range of experimental conditions such as inhomogeneous rf fields, spread of chemical shifts, the intrinsic orientation dependencies of powder samples, and sample spinning. While the vast majority of previous dipolar recoupling sequences are operating through planar double-or zero-quantum effective Hamiltonians, we present here not only improved variants of such experiments but also for the first time homonuclear isotropic mixing sequences which transfers all I(x), I(y), and I(z) polarizations from one spin to the same operators on another spin simultaneously and with equal efficiency. This property may be exploited to increase the signal-to-noise ratio of two-dimensional experiments by a factor of square root 2 compared to conventional solid-state methods otherwise showing the same efficiency. The sequences are tested numerically and experimentally for a powder of (13)C(alpha),(13)C(beta)-L-alanine and demonstrate substantial sensitivity gains over previous dipolar recoupling experiments. PMID- 17115761 TI - Molecular dynamics investigation with the time resolved optical Kerr effect on the CS2-C6H6 mixtures. AB - An investigation of the molecular dynamics in pure liquids and in mixtures through the technique of time resolved optical Kerr effect is performed. The samples studied were the mixtures of carbon disulfide (CS(2)) with benzene (C(6)H(6)). The molecular dynamics of the pure liquids is briefly discussed while the main results are obtained for the mixtures. A slow dynamics component is observed for the optical heterodyne detected optical Kerr effect transient decaying exponentially with time constants on picosecond time scale. The fast subpicosend time relaxations are analyzed in terms of the nondiffusive component of the spectral response that is associated with the molecular dynamics. The modifications of the spectrum are quantified, and the explanation of the observed changes is given in terms of the structural interaction configurations that produced changes in the intermolecular potential within which the molecules execute librational motions. PMID- 17115762 TI - Melting temperature: from nanocrystalline to amorphous phase. AB - By extrapolating the mean grain size of nanocrystal to an infinitesimal value, an amorphous phase has been obtained from the Voronoi construction. The molecular dynamics simulations indicated that for nanocrystal, the grain size variation of melting temperature exhibits two characteristic regions. As mean grain size above about 4 nm for Ag, the melting temperatures decrease with decreasing grain size. However, with grain size further shrinking, the melting temperatures almost keep a constant. This is because the dominant factor on the melting temperature of nanocrystal shifts from grain phase to grain boundary. As a result of fundamental difference in structure, the amorphous phase has a much lower solid-to-liquid transformation temperature than that of nanocrystal. PMID- 17115763 TI - Structural relaxation in complex liquids: non-Markovian dynamics in a bistable potential. AB - The time correlation function C(t) identical with of the distance fluctuations of a particle moving in a bistable potential under the action of fractional Gaussian noise (fGn) is calculated from a Smoluchowski-type equation derived from a generalized Langevin equation (GLE). The time derivative of this function, dC(t)dt, is compared with data from optical Kerr effect measurements of liquid crystal dynamics in the vicinity of the isotropic-to-nematic transition, which are related to the time derivative of an orientational correlation function. A number of characteristic features of the experimental decay curves, including short and intermediate time power law behavior and long time exponential relaxation, are qualitatively reproduced by the analytical calculations, even though the latter do not explicitly treat orientational degrees of freedom. The GLE formalism with fGn was, in fact, originally proposed as a model of protein conformational fluctuations, so the present results suggest that it may also serve more generally as a model of structural relaxation in complex condensed phase media. PMID- 17115764 TI - Computational study of structural and dynamical properties of formamide-water mixtures. AB - A molecular dynamics simulation study of structural and dynamical properties in liquid mixtures of formamide and water is presented. Site-site radial pair distribution functions, local mole fractions, pair energy distributions, and tetrahedral orientational order are the quantities analyzed to investigate the local structure in the simulated mixtures, along with a review of the intermolecular structure in terms of the distribution of hydrogen bonds. Our results indicate that there is a substitution of formamide molecules by water in the hydrogen bonds and a formation of a common hydrogen bond network. By analyzing the extent of tetrahedral order in the liquid as a function of composition, it is observed that whereas the tetrahedral network of liquid water is progressively lost by increasing the formamide concentration, the water structure within the first coordination shell is preserved and somewhat enhanced. The hydrogen-bond mean lifetimes were estimated by performing a time integration of the autocorrelation functions of bond occupation numbers. The lifetimes associated with hydrogen bonds between water, formamide, and interspecies pairs are found to increase with increasing formamide concentration. The lifetimes of the water hydrogen bonds show the largest variations, supporting the picture of an enhancement of the water structure among the nearest neighbors within the first coordination shell. We have used two different force field models for water, SPC/E [J. C. Berendsen et al., J. Phys. Chem. 91, 6269 (1987)] and TIP4P/2005 [J. L. F. Abascal and C. Vega, J. Chem. Phys. 123, 234505 (2005)]. Our results for structural and dynamical properties yield very small differences between those models, the TIP4P/2005 predicting a slightly more structured liquid and, consequently, exhibiting a slightly slower translational and librational dynamics. PMID- 17115765 TI - An accurate and simple quantum model for liquid water. AB - The path-integral molecular dynamics and centroid molecular dynamics methods have been applied to investigate the behavior of liquid water at ambient conditions starting from a recently developed simple point charge/flexible (SPC/Fw) model. Several quantum structural, thermodynamic, and dynamical properties have been computed and compared to the corresponding classical values, as well as to the available experimental data. The path-integral molecular dynamics simulations show that the inclusion of quantum effects results in a less structured liquid with a reduced amount of hydrogen bonding in comparison to its classical analog. The nuclear quantization also leads to a smaller dielectric constant and a larger diffusion coefficient relative to the corresponding classical values. Collective and single molecule time correlation functions show a faster decay than their classical counterparts. Good agreement with the experimental measurements in the low-frequency region is obtained for the quantum infrared spectrum, which also shows a higher intensity and a redshift relative to its classical analog. A modification of the original parametrization of the SPC/Fw model is suggested and tested in order to construct an accurate quantum model, called q-SPC/Fw, for liquid water. The quantum results for several thermodynamic and dynamical properties computed with the new model are shown to be in a significantly better agreement with the experimental data. Finally, a force-matching approach was applied to the q-SPC/Fw model to derive an effective quantum force field for liquid water in which the effects due to the nuclear quantization are explicitly distinguished from those due to the underlying molecular interactions. Thermodynamic and dynamical properties computed using standard classical simulations with this effective quantum potential are found in excellent agreement with those obtained from significantly more computationally demanding full centroid molecular dynamics simulations. The present results suggest that the inclusion of nuclear quantum effects into an empirical model for water enhances the ability of such model to faithfully represent experimental data, presumably through an increased ability of the model itself to capture realistic physical effects. PMID- 17115766 TI - Chemistry, photophysics, and ultrafast kinetics of two structurally related Schiff bases containing the naphthalene or quinoline ring. AB - The two structurally related Schiff bases, 2-hydroxynaphthylidene-(8 aminoquinoline) (HNAQ) and 2-hydroxynaphthylidene-1(')-naphthylamine (HNAN), were studied by means of steady-state and time resolved optical spectroscopies as well as time-dependent density functional theory (TDDFT) calculations. The first one, HNAQ, is stable as a keto tautomer in the ground state and in the excited state in solutions, therefore it was used as a model of a keto tautomer of HNAN which exists mainly in its enol form in the ground state at room temperature. Excited state intramolecular proton transfer in the HNAN molecule leads to a very weak (quantum yield of the order of 10(-4)) strongly Stokes-shifted fluorescence. The characteristic time of the proton transfer (about 30 fs) was estimated from femtosecond transient absorption data supported by global analysis and deconvolution techniques. Approximately 35% of excited molecules create a photochromic form whose lifetime was beyond the time window of the experiment (2 ns). The remaining ones reach the relaxed S(1) state (of a lifetime of approximately 4 ps), whose emission is present in the decay associated difference spectra. Some evidence for the back proton transfer from the ground state of the keto form with the characteristic time of approximately 13 ps was also found. The energies and orbital characteristics of main electronic transitions in both molecules calculated by TDDFT method are also discussed. PMID- 17115767 TI - Space-time thermodynamics and subsystem observables in a kinetically constrained model of glassy materials. AB - In a recent article [M. Merolle et al., Proc. Natl. Acad. Sci. U.S.A. 102, 10837 (2005)], it was argued that dynamic heterogeneity in d-dimensional glass formers is a manifestation of an order-disorder phenomenon in the d+1 dimensions of space time. By considering a dynamical analog of the free energy, evidence was found for phase coexistence between active and inactive regions of space time, and it was suggested that this phenomenon underlies the glass transition. Here we develop these ideas further by investigating in detail the one-dimensional Fredrickson-Andersen (FA) model, in which the active and inactive phases originate in the reducibility of the dynamics. We illustrate the phase coexistence by considering the distributions of mesoscopic space-time observables. We show how the analogy with phase coexistence can be strengthened by breaking microscopic reversibility in the FA model, leading to a nonequilibrium theory in the directed percolation universality class. PMID- 17115768 TI - Neutron diffraction data and molecular dynamics simulations of the molten mixture Ag(Br0.7I0.3). AB - The structure factors of the ionic liquid mixture Ag(Br(0.7)I(0.3)) at three temperatures, 723, 923, and 1023 K, as well as of the pure molten AgI at 923 K and the pure molten AgBr at 773 and 923 K, were studied experimentally and by means of molecular dynamics simulations. The experiments were carried out using the high intensity total scattering time-of-flight spectrometer, HIT-II, at the KENS spallation neutron source in Japan. The experimental data are very reliable, with the possible exception of the small momentum transfer region, whose accessibility is limited by neutron energy and detector positions. The simulations made use of the semiempirical rigid ion potentials of the Vashishta Rahman [Phys. Rev. Lett. 40, 1337 (1978)] type using a new set of parameters appropriate for the mixture. Within the known constraints of the pairwise rigid ion potentials, the simulated structure factors are in fair agreement with experiment. The results for the pair distribution functions suggest that the molten mixture retains the superionic character found in previous calculations of both the AgI and AgBr melts. This suggestion is confirmed by the results for the self-diffusion coefficients. Values obtained for the ionic conductivities are also presented. PMID- 17115769 TI - The Prigogine-Defay ratio revisited. AB - One of the basic characteristics of the glass transition, the Prigogine-Defay ratio, connecting jumps of the thermal expansion coefficient, isothermal compressibility, and isobaric specific heat capacity in vitrification is rederived in the framework of the thermodynamics of irreversible processes employing the order-parameter concept introduced by de Donder and van Rysselberghe [Thermodynamic Theory of Affinity (Stanford University Press, Stanford, 1936)]. In our analysis, glass-forming liquids and glasses are described by only one structural order parameter. However, in contrast to previous approaches to the derivation of this ratio, the process of vitrification is treated not in terms of Simon's simplified model [Z. Anorg. Allg. Chem. 203, 219 (1931)] as a freezing-in process proceeding at some sharp temperature, the glass transition temperature T(g), but in some finite temperature interval accounting appropriately for the nonequilibrium character of vitrifying systems in this temperature range. As the result of the theoretical analysis, we find, in particular, that the Prigogine-Defay ratio generally has to have values larger than 1 for vitrification in cooling processes. Quantitative estimates of the Prigogine-Defay ratio are given utilizing a mean-field lattice-hole model of glass-forming melts. Some further consequences are derived concerning the behavior of thermodynamic coefficients, in particular, of Young's modulus in vitrification. The theoretical results are found to be in good agreement with experimental data. PMID- 17115770 TI - Phase separation and percolation of reversibly aggregating spheres with a square well attraction potential. AB - Reversible aggregation of spheres is simulated using a novel method in which clusters of bound spheres diffuse collectively with a diffusion coefficient proportional to their radius. It is shown that the equilibrium state is the same as with other simulation techniques, but with the present method more realistic kinetics are obtained. The behavior as a function of volume fraction and interaction strength was tested for two different attraction ranges. The binodal and the percolation threshold were determined. The cluster structure and size distribution close to the percolation threshold were found to be consistent with the percolation model. Close to the binodal phase separation occurred through the growth of spherical dense domains, while for deep quenches a system spanning network is formed that coarsens with a rate that decreases with increasing attraction. We found no indication for arrest of the coarsening. PMID- 17115771 TI - The dynamical properties of the aromatic hydrogen bond in NH4(C6H5)4B from quasielastic neutron scattering. AB - NH(4)(C(6)H(5))(4)B represents a prototypical system for understanding aromatic H bonds. In NH(4)(C(6)H(5))(4)B an ammonium cation is trapped in an aromatic cage of four phenyl rings and each phenyl ring serves as a hydrogen bond acceptor for the ammonium ion as donor. Here the dynamical properties of the aromatic hydrogen bond in NH(4)(C(6)H(5))(4)B were studied by quasielastic incoherent neutron scattering in a broad temperature range (20< or =T< or =350 K). We show that in the temperature range from 67 to 350 K the ammonium ions perform rotational jumps around C(3) axes. The correlation time for this motion is the lifetime of the "transient" H bonds. It varies from 1.5 ps at T=350 K to 150 ps at T=67 K. The activation energy was found to be 3.14 kJ mol, which means only 1.05 kJ mol per single H bond for reorientations around the C(3) symmetry axis of the ammonium group. This result shows that the ammonium ions have to overcome an exceptionally low barrier to rotate and thereby break their H bonds. In addition, at temperatures above 200 K local diffusive reorientational motions of the phenyl rings, probably caused by interaction with ammonium-group reorientations, were found within the experimental observation time window. At room temperature a reorientation angle of 8.4 degrees +/-2 degrees and a correlation time of 22+/-8 ps were determined for the latter. The aromatic H bonds are extremely short lived due to the low potential barriers allowing for molecular motions with a reorientational character of the donors. The alternating rupture and formation of H bonds causes very strong damping of the librational motion of the acceptors, making the transient H bond appear rather flexible. PMID- 17115772 TI - On theoretical predictions of noble-gas hydrides. AB - We discuss the present status and reliability of theoretical predictions of noble gas hydride molecules. It is shown that the single-reference MP2 calculations can produce a rather inaccurate energy diagram for the formation of noble-gas hydrides, and this may mislead the theoretical predictions. We suggest that the computational dissociation energy of the HY precursors should always be compared with the experimental values as a checkpoint for the computational accuracy. The computational inaccuracy probably explains why some compounds that are stable with the single-reference MP2 method (HArC(4)H, HArC(3)N, and HArCN) did not appear in matrix-isolation experiments, whereas the corresponding compounds with Kr and Xe are known. PMID- 17115773 TI - Microscopic theory for interface fluctuations in binary liquid mixtures. AB - Thermally excited capillary waves at fluid interfaces in binary liquid mixtures exhibit simultaneously both density and composition fluctuations. Based on a density functional theory for inhomogeneous binary liquid mixtures we derive an effective wavelength dependent Hamiltonian for fluid interfaces in these systems beyond the standard capillary-wave model. Explicit expressions are obtained for the surface tension, the bending rigidities, and the coupling constants of compositional capillary waves in terms of the profiles of the two number densities characterizing the mixture. These results lead to predictions for grazing-incidence x-ray scattering experiments at such interfaces. PMID- 17115774 TI - The symmetry of single-molecule conduction. AB - We introduce the conductance point group which defines the symmetry of single molecule conduction within the nonequilibrium Green's function formalism. It is shown, either rigorously or to within a very good approximation, to correspond to a molecular-conductance point group defined purely in terms of the properties of the conducting molecule. This enables single-molecule conductivity to be described in terms of key qualitative chemical descriptors that are independent of the nature of the molecule-conductor interfaces. We apply this to demonstrate how symmetry controls the conduction through 1,4-benzenedithiol chemisorbed to gold electrodes as an example system, listing also the molecular-conductance point groups for a range of molecules commonly used in molecular electronics research. PMID- 17115775 TI - Site-directed electronic tunneling through a vibrating molecular network. AB - The effect of electronic-nuclear coupling on electronic transport through a complex molecular network is studied. Electronic tunneling dynamics in a network of N donor/acceptor sites, connected by molecular bridges, is shown to be controlled by electronic-nuclear coupling at the bridges. Particularly, electronic coupling to an accepting nuclear mode at the contact site between the donor and the rest of the network is shown to affect the tunneling path selection to specific acceptors. In the "deep" tunneling regime, the network is mapped onto an N-level system using a recursive perturbation expansion, enabling analytical treatment of the electronic dynamics. The analytic formulation is applied for two model systems, demonstrating site-directed tunneling by electronic-nuclear coupling. Numerical simulations suggest that this phenomenon is not limited to the deep tunneling regime. PMID- 17115776 TI - Surface properties of fluids of charged platelike colloids. AB - Surface properties of mixtures of charged platelike colloids and salt in contact with a charged planar wall are studied within density functional theory. The particles are modeled by hard cuboids with their edges constrained to be parallel to the Cartesian axes corresponding to the Zwanzig model [J. Chem. Phys. 39, 1714 (1963)] and the charges of the particles are concentrated at their centers. The density functional applied is an extension of a recently introduced functional for charged platelike colloids. It provides a qualitative approach because it does not determine the relation between the actual and the effective charges entering into the model. Technically motivated approximations, such as using the Zwanzig model, are expected not to influence the results qualitatively. Analytically and numerically calculated bulk and surface phase diagrams exhibit first-order wetting for sufficiently small macroion charges and isotropic bulk order as well as first-order drying for sufficiently large macroion charges and nematic bulk order. The asymptotic wetting and drying behaviors are investigated by means of effective interface potentials which turn out to be asymptotically the same as for a suitable neutral system governed by isotropic nonretarded dispersion forces. Wetting and drying points as well as predrying lines and the corresponding critical points have been located numerically. A crossover from monotonic to nonmonotonic electrostatic potential profiles upon varying the surface charge density has been observed. Nonmonotonic electrostatic potential profiles are equivalent to the occurrence of charge inversion. Due to the presence of both the Coulomb interactions and the hard-core repulsions, the surface potential and the surface charge do not vanish simultaneously, i.e., the point of zero charge and the isoelectric point of the surface do not coincide. PMID- 17115777 TI - The environmental effect on the radial breathing mode of carbon nanotubes. II. Shell model approximation for internally and externally adsorbed fluids. AB - We have previously shown that the upshift in the radial breathing mode (RBM) of closed (or infinite) carbon nanotubes in solution is almost entirely due to coupling of the RBM with an adsorbed layer of fluid on the nanotube surface. The upshift can be modeled analytically by considering the adsorbed fluid as an infinitesimally thin shell, which interacts with the nanotube via a continuum Lennard-Jones potential. Here we extend the model to include internally as well as externally adsorbed waterlike molecules, and find that filling the nanotubes leads to an additional upshift of two to six wave numbers. We show that using molecular dynamics, the RBM can be accurately reproduced by replacing the fluid molecules with a mean field harmonic shell potential, greatly reducing simulation times. PMID- 17115778 TI - Accelerated molecular dynamics simulation of the thermal desorption of n-alkanes from the basal plane of graphite. AB - We utilize accelerated molecular dynamics to simulate alkane desorption from the basal plane of graphite. Eight different molecules, ranging from n-pentane to n hexadecane, are studied in the low coverage limit. Acceleration of the molecular dynamics simulations is achieved using two different methods: temperature acceleration and a compensating potential scheme. We find that the activation energy for desorption increases with increasing chain length. The desorption prefactor increases with chain length for molecules ranging from pentane to decane. This increase subsides and the value of the preexponential factor fluctuates about an apparently constant value for decane, dodecane, tetradecane, and hexadecane. These trends are consistent with data obtained in experimental temperature-programed desorption (TPD) studies. We explain the dependence of the preexponential factor on alkane chain length by examining conformational changes within the alkane molecules. For the shorter molecules, torsional motion is not activated over experimental temperature ranges. These molecules can be treated as rigid rods and their partial loss in translational and rotational entropies upon adsorption increases as chain length increases, leading to an increasing preexponential factor. At their typical TPD peak temperatures, torsions are activated in the longer adsorbed chain molecules to a significant extent which increases with increasing chain length, increasing the entropy of the adsorbed molecule. This increase counteracts the decrease in entropy due to a loss of translation and rotation, leading to a virtually constant prefactor. PMID- 17115779 TI - Surface order-disorder phase transitions and percolation. AB - In the present paper, the connection between surface order-disorder phase transitions and the percolating properties of the adsorbed phase has been studied. For this purpose, four lattice-gas models in the presence of repulsive interactions have been considered. Namely, monomers on honeycomb, square, and triangular lattices, and dimers (particles occupying two adjacent adsorption sites) on square substrates. By using Monte Carlo simulation and finite-size scaling analysis, we obtain the percolation threshold theta(c) of the adlayer, which presents an interesting dependence with w/k(B)T (w, k(B), and T being the lateral interaction energy, the Boltzmann constant, and the temperature, respectively). For each geometry and adsorbate size, a phase diagram separating a percolating and a nonpercolating region is determined. PMID- 17115780 TI - Submolecular imaging of chloronitrobenzene isomers on Cu(111). AB - We compare computer simulations to experimental scanning tunneling microscopy (STM) images of chloronitrobenzene molecules on a Cu(111) surface. The experiments show that adsorption induced isomerization of the molecules takes place on the surface. Furthermore, not only the submolecular features can be seen in the STM images, but different isomers can also be recognized. The Todorov Pendry approach to tunneling produces simulated STM images which are in good accordance with the experiments. Alongside with STM simulations in a tight binding basis, ab initio calculations are performed in order to analyze the symmetry of relevant molecular orbitals and to consider the nature of tunneling channels. Our calculations show that while the orbitals delocalized to the phenyl ring create a relatively transparent tunneling channel, they also almost isolate the orbitals of the substitute groups at energies which are relevant in STM experiments. These features of the electronic structure are the key ingredients of the accurate submolecular observations. PMID- 17115781 TI - Exciton-phonon coupling and disorder in the excited states of CdSe colloidal quantum dots. AB - We study the origin of the spectral line shape in colloidal CdSe nanocrystal quantum dots. The three-pulse photon echo peak shift (3PEPS) data reveal a temperature-independent fast decay, obscuring the quantification of the homogeneous linewidth. The optical gap and Stokes shift are found to have an anomalous behavior with temperature, which is size, capping group, and surrounding polymer matrix independent. Using these results and combining them with simulations, we discuss the role of exciton-phonon coupling, static inhomogeneity, exciton fine structure, and exciton state disorder in the linewidth of the nanocrystal. In particular, our analysis shows that the disorder due to surface imperfections and finite temperature effects, as well as the relaxation within the fine structure, can have significant impact on the steady state absorption spectrum, 3PEPS data, and dephasing processes. PMID- 17115782 TI - Coexistence of one- and two-dimensional supramolecular assemblies of terephthalic acid on Pd(111) due to self-limiting deprotonation. AB - The adsorption of terephthalic acid [C(6)H(4)(COOH)(2), TPA] on a Pd(111) surface has been investigated by means of scanning tunneling microscopy (STM), x-ray photoelectron spectroscopy, and near-edge x-ray absorption fine structure spectroscopy under ultrahigh vacuum conditions at room temperature. We find the coexistence of one- (1D) and two-dimensional (2D) molecular ordering. Our analysis indicates that the 1D phase consists of intact TPA chains stabilized by a dimerization of the self-complementary carboxyl groups, whereas in the 2D phase, consisting of deprotonated entities, the molecules form lateral ionic hydrogen bonds. The supramolecular growth dynamics and the resulting structures are explained by a self-limiting deprotonation process mediated by the catalytic activity of the Pd surface. Our models for the molecular ordering are supported by molecular mechanics calculations and a simulation of high resolution STM images. PMID- 17115783 TI - Molecular dynamics simulations of surface-initiated melting of nitromethane. AB - The melting of nitromethane initiated at solid-vacuum interfaces has been investigated using molecular dynamics nvt simulations with a realistic force field [D. C. Sorescu et al., J. Phys. Chem. B 104, 8406 (2000)]. The calculated melting point (251+/-5 K) is in good agreement with experiment (244.73 K) and values obtained previously (approximately 255.5 and 266.5+/-8 K) using other simulation methods [P. M. Agrawal et al., J. Chem. Phys. 119, 9617 (2003)]. Analyses of the molecular orientations and diffusion during the simulations as functions of the distance from the exposed surfaces show that the melting is a direct crystal-to-liquid transition, in which the molecules first gain rotational freedom, then mobility. There is a slight dependence of the melting temperature on the exposed crystallographic face. PMID- 17115784 TI - Second harmonic generation from small gold metallic particles: from the dipolar to the quadrupolar response. AB - Hyper Raleigh scattering, a common technique to investigate the second harmonic light scattered from a liquid suspension of molecular compounds and to determine their quadratic hyperpolarizability, has been used for aqueous suspensions of gold nanoparticles, the diameter of which ranges from 20 up to 150 nm. The hyper Rayleigh signal intensity was recorded as a function of the angle of polarization of the incident fundamental wave. For the particles with a diameter smaller than 50 nm, the response is dominated by the dipolar contribution arising from the deviation of the particle shape from that of a perfect sphere. For larger diameter particles, retardation effects in the interaction of the electromagnetic fields with the particles cannot be neglected any longer and the response deviates from the pure dipolar response, exhibiting a strong quadrupolar contribution. It is then shown that in order to quantify the relative magnitude of these two dipolar and quadrupolar contributions, a weighting parameter zeta(V) which equals unity for a pure quadrupolar contribution and vanishes for a pure dipolar response, can be introduced. PMID- 17115785 TI - Single quintuple bond [PhCrCrPh] molecule as a possible molecular switch. AB - The electronic transport properties of a single quintuple bond [PhCrCrPh] molecule sandwiched between two Au(111) surfaces with the trans-bent and linear configurations are studied by a fully self-consistent nonequilibrium Green's function method combined with density functional theory. The calculated transmission spectra of two chemical isomers are remarkably distinctive. Theoretical results suggest that the current through the trans-bent configuration is significantly larger than the corresponding linear one. The predicted on-off ratio of currents ranging from around 50 to 200 in the applied bias window [-1.5 V, 1.5 V] suggests that multiple bond compounds have attractive potential in molecular switch technology. PMID- 17115786 TI - Achieving optimum hydrogen permeability in PdAg and PdAu alloys. AB - The present work investigates both the diffusivity and permeability of hydrogen (H) in palladium-silver (PdAg) and palladium-gold (PdAu) alloys over a 400-1200 K temperature range for Pd(100-X)M(X), M=Ag or Au and X=0%-48% using density functional theory (DFT) and kinetic Monte Carlo simulations (KMC). DFT has been employed to obtain octahedral (O)-, tetrahedral (T)-, and transition state (TS)- site energetics as a function of local alloy composition for several PdAg and PdAu alloys with compositions in supercells of X=14.18%, 25.93%, 37.07%, and 48.15% with the nearest (NNs) and next nearest neighbors (NNNs) varied over the entire range of compositions. The estimates were then used to obtain a model relating the O, T, and TS energies of a given site with NN(X), NNN(X), and the lattice constant. The first passage approach combined with KMC simulations was used for the H diffusion coefficient predictions. It was found that the diffusion coefficient of H in PdAg alloy decreases with increasing Ag and increases with increasing temperature, matching closely with the experimental results reported in the literature. The calculated permeabilities of H in these novel binary alloys obtained from both diffusivity and solubility predictions were found to have a maximum at approximately 20% Ag and approximately 12% Au, which agree well with experimental predictions. Specifically, the permeability of H in PdAg alloy with approximately 20% Ag at 456 K is three to four times that of pure Pd, while the PdAu alloy at 12% Au is four to five times that of pure Pd at 456 K. PMID- 17115787 TI - Three-particle correlation functions of quasi-two-dimensional one-component and binary colloid suspensions. AB - We report the results of experimental determinations of the triplet correlation functions of quasi-two-dimensional one-component and binary colloid suspensions in which the colloid-colloid interaction is short ranged. The suspensions studied range in density from modestly dilute to solid. The triplet correlation function of the one-component colloid system reveals extensive ordering deep in the liquid phase. At the same density the ordering of the larger diameter component in a binary colloid system is greatly diminished by a very small amount of the smaller diameter component. The possible utilization of information contained in the triplet correlation function in the theory of melting of a quasi-two-dimensional system is briefly discussed. PMID- 17115788 TI - Charge driven, electrohydrodynamic patterning of thin films. AB - In electrohydrodynamic patterning, electrical forces and surface tension acting at the interface between two fluids sandwiched between silicon wafers compete to set the period of pillar arrays, gratings, and concentric rings. Shrinking the period to deep submicron lengths requires a precise understanding of the source of the electric field. Previous modeling efforts have assumed that applied voltages, contact potentials, and static charge drive the flow. Here we show the location of that charge and the tangential stress it engenders to impact profoundly how the period and growth rate depend on the dielectric contrast and the relative film thickness. The pillar-to-pillar spacing scales inversely proportional to the charge density, and densities of approximately 1 mC/m(2) (approximately 1 charge/100 nm(2)) suffice to produce micron sized pillars. PMID- 17115789 TI - Multiple short time power laws in the orientational relaxation of nematic liquid crystals. AB - Relaxation in the nematic liquid crystalline phase is known to be sensitive to its proximity to both isotropic and smectic phases. Recent transient optical Kerr effect (OKE) studies have revealed, rather surprisingly, two temporal power laws at short to intermediate times and also an apparent absence of the expected exponential decay at longer times. In order to understand this unusual dynamics, we have carried out extensive molecular dynamics simulations of transient OKE and related orientational time correlation functions in a system of prolate ellipsoids (with aspect ratio equal to 3). The simulations find two distinct power laws, with a crossover region, in the decay of the orientational time correlation function at short to intermediate times (in the range of a few picoseconds to a few nanoseconds). In addition, the simulation results fail to recover any long time exponential decay component. The system size dependence of the exponents suggests that the first power law may originate from the local orientational density fluctuations (like in a glassy liquid). The origin of the second power law is less clear and may be related to the long range fluctuations (such as smecticlike density fluctuations)--these fluctuations are expected to involve small free energy barriers. In support of the latter, the evidence of pronounced coupling between orientational and spatial densities at intermediate wave numbers is presented. This coupling is usually small in normal isotropic liquids, but it is large in the present case. In addition to slow collective orientational relaxation, the single particle orientational relaxation is also found to exhibit slow dynamics in the nematic phase in the long time. PMID- 17115790 TI - Schmidt number effects in dissipative particle dynamics simulation of polymers. AB - Simulation studies for dilute polymeric systems are presented using the dissipative particle dynamics method. By employing two different thermostats, the velocity-Verlet and Lowe's scheme, we show that the Schmidt number (S(c)) of the solvent strongly affects nonequilibrium polymeric quantities. The fractional extension of wormlike chains subjected to steady shear is obtained as a function of S(c). Poiseuille flow in microchannels for fixed polymer concentration and varying number of repeated units within a chain is simulated. The nonuniform concentration profiles and their dependence on S(c) are computed. We show the effect of the bounce-forward wall boundary condition on the depletion layer thickness. A power law fit of the velocity profile in stratified Poiseuille flow in a microchannel yields wall viscosities different from bulk values derived from uniform, steady plane Couette flow. The form of the velocity profiles indicates that the slip flow model is not useful for the conditions of these calculations. PMID- 17115791 TI - Simulation studies of self-assembly of end-tethered nanorods in solution and role of rod aspect ratio and tether length. AB - We present temperature versus concentration phase diagrams for "shape amphiphiles" comprised of tethered moderate and low aspect ratio rods. Simulations of moderate aspect ratio rods (first reported by Horsch et al. [Phys. Rev. Lett. 95, 056105 (2005)]) predict their self-assembly into spherical micelles with bcc order, long micelles with nematic order, a racemic mixture of hexagonally ordered chiral cylinders, two perforated phases: one with tetragonal order and one with hexagonal order, and a smectic C lamellar phase. In contrast, we predict here that small aspect ratio tethered rods self-assemble into bcc ordered spherical micelles, hexagonally ordered cylinders, and a smectic C lamellar phase. We compare and contrast the phases obtained for the two aspect ratios and examine in further detail several unusual phases. Our simulations also reveal that for moderate aspect ratio rods there is a tendency toward phases with decreasing interfacial curvature with decreasing coil size, including a double gyroid phase. In addition, we investigate the role of tether length on the assembled structures. Our results are applicable to short rod-coil block copolymers and rodlike nanoparticles with polymer tethers, and to colloidal building blocks comprised of a flexible string of colloids tethered to a rigid string of colloids, with the interactions scaled appropriately. PMID- 17115792 TI - Single chain in mean field simulations: quasi-instantaneous field approximation and quantitative comparison with Monte Carlo simulations. AB - The description of fluctuations by single chain in mean field (SCMF) simulations is discussed and the results of this particle-based self-consistent field technique are quantitatively compared to Monte Carlo simulations of the same discretized Edwards-Hamiltonian providing exact reference data. In SCMF simulations one studies a large ensemble of noninteracting molecules subjected to real, external fields by Monte Carlo simulations. The external fields approximate nonbonded, instantaneous interactions between molecules. In the self-consistent mean field theory the external fields are static and fluctuation effects are ignored. In SCMF simulations, the external fields fluctuate since they are frequently recalculated from the instantaneous density distribution of the ensemble of molecules. In the limit of infinitely high density or instantaneous update of the external fields, the SCMF simulation method accurately describes long-wavelength fluctuations. At high but finite updating frequency the accuracy depends on the discretization of the model. The accuracy is illustrated by studying the single chain structure and intermolecular correlations in polymer melts, and fluctuation effects on the order-disorder transition of symmetric diblock copolymers. PMID- 17115793 TI - Minimal model of relaxation in an associating fluid: viscoelastic and dielectric relaxations in equilibrium polymer solutions. AB - Cluster formation and disintegration greatly complicate the description of relaxation processes in complex fluids. We systematically contrast the viscoelastic and dielectric properties for models of equilibrium polymers whose thermodynamic properties have previously been established. In particular, the monomer-mediated model allows chain growth to proceed only by monomer addition, while the scission-recombination model enables all particles to associate democratically, so that chain scission and fusion occur at the interior segments as well as at chain ends. The minimal models neglect hydrodynamic and entanglement interactions and are designed to explore systematically the competition between chemical reaction and internal chain relaxation and how this coupling modifies the dynamics from that of a polydisperse solution of Rouse chains with fixed lengths (i.e., "frozen" chains). As expected, the stress relaxation is nearly single exponential when the assembly-disassembly reaction is fast on the time scale of structural chain rearrangements, while multiexponential or nearly stretched exponential relaxation is obtained when this reaction rate is slow compared to the broad relaxation spectrum of almost unperturbed, nearly "dead" chains of intrinsically polydisperse equilibrium polymer solutions. More generally, a complicated intermediate behavior emerges from the interplay between the chemical kinetic events and internal chain motions. PMID- 17115794 TI - An improved collision efficiency model for particle aggregation. AB - A generalized geometric model is presented which describes the collision efficiency factor of aggregation (the probability of a binary particle or aggregate collision resulting in adhesion) for systems comprised of two oppositely charged species. Application of the general model to specific systems requires calculation of the area of each species available for collision with a second species. This is in contrast to previous models developed for polymer particle flocculation that are based on the fractional surface coverage of adsorbed polymer. The difference between these approaches is suggested as an explanation for previously observed discrepancies between theory and observation. In the current work the specific case of oppositely charged nondeformable spherical particles (heteroaggregation) is quantitatively addressed. The optimum concentration of oppositely charged particles for rapid aggregation (maximum collision efficiency) as a function of relative particle size is calculated and an excellent correlation is found with data taken from literature. PMID- 17115795 TI - Local polymer dynamics under strong connectivity constraints: the dendrimer case. AB - The characteristics of local motion are explored by molecular dynamics simulations in a series of AB(2)-type dendrimer melts. Systems of generations 3-5 were simulated in a wide temperature range, allowing the assessment of effects associated with molecular size, proximity to the detected glasslike transitions, and the strong connectivity constraints imposed by the dendritic topology. Investigation of the mechanisms involved in local motion at short temporal and spatial scales revealed the connection between the non-Gaussian nature of monomer displacements to alpha-relaxation and the caging/decaging process under different degrees of confinement. In the latter mechanism, two characteristic localization lengths were identified: at the low temperature limit spatial localization was realized within approximately 10% of the nearest neighbor distance while at temperatures higher than the glass transition, the existence of an analogous length scale is ascribed to the geometric constraints due to the dense connectivity pattern. As the results from this study are discussed in comparison to the behavior observed in linear polymers and supercooled liquids, new insight is provided on the universal/specific mechanisms involved in local dynamics of different glass-forming systems. PMID- 17115796 TI - Darkness at noon: sunscreens and vitamin D3. AB - We are assured by responsible scientific and governmental organizations that sunscreens should be routinely worn to reduce skin cancer risk. We are also advised that wearing sunscreens will not hinder our ability to produce sufficient previtamin D3 (preD3) from casual sunlight exposure. We report the examination of a series of 166 solar spectra, obtained on different days throughout a year, evaluated for erythemic and preD3 effectiveness and the relative effects of recommended Sun protection factor (SPF) 15 sunscreen. The results show that the sunscreen is much more effective in blocking the formation of preD3, than its labeled SPF for preventing sunburn. In fact with sunscreen applied only miniscule amounts of preD3 are predicted to be made outdoors even with extensive exposure. This raises important questions regarding the safest way to use sunlight exposure to promote healthy vitamin D3 levels and suggests the need to modify the public safety "Safe Sun" messages. PMID- 17115797 TI - Effect of solvent polarity and viscosity on the guest binding dynamics with bile salt aggregates. AB - Bile salts form supramolecular aggregates with two binding sites with different properties. The guest binding dynamics to the aggregates and guest protection from species in the aqueous phase were investigated using fluorescence and laser flash photolysis experiments. Sodium cholate, deoxycholate and taurodeoxycholate were used as bile salts and acetonitrile or ethylene glycol were added as co solvents to water in order to alter the binding properties of 1-ethylnaphthalene and 1-naphthyl-1-ethanol with the aggregates. The binding dynamics are faster and protection efficiencies are lower for guests bound to cholate and in the presence of either co-solvent. PMID- 17115798 TI - Visual functional effects of constant blue light in a retinal degenerate rat model. AB - Retinal degenerative conditions increase susceptibility to light damage, but rapid retinal degeneration (RD) models show less susceptibility to cyclic dim light. We investigated whether constant blue light (BL) exposure can eliminate the residual visual responses in a comparatively rapid RD rat model. Pigmented rhodopsin mutant S334ter line-3 rat pups (21 days old) were exposed for 5-6 consecutive days to constant BL. Visual behavior was evaluated with an optokinetic head tracking apparatus. Electrophysiological recordings were made from the superior colliculus (SC). S-antigen, red-green opsin and rhodopsin immunoreactive residual photoreceptors were counted. Following BL exposure, head tracking was significantly reduced at 0.25 cycles degree(-1) in 38-day-old line 3 rats. With a 0.125 cycles degree(-1) stimulus, the head tracking performance of 80-day-old BL rats were similar to that of 220-day-old no-BL-treated line-3 rats. SC recordings also revealed a significant decrease in the residual photoreceptor activity. Histological evaluation showed reduction of the rod population in the central area of the light-damaged retina. Exposure to constant BL considerably reduces the residual visual responses in a rapid degenerating RD rat model. PMID- 17115799 TI - Sensitivity of erythemally effective UV irradiance and daily exposure to uncertainties in measured total ozone. AB - In this study the sensitivity of the erythemally effective radiation to uncertainties in operationally measured total ozone content of the atmosphere (TOC) was estimated. For this, daily operational TOC measurements from different instruments were applied covering the period from 1997 to 1999. Measurements were gained from space by Earth Probe Satellite, Earth Remote Sensing satellite/Global Ozone Monitoring Experiment and Operational Vertical Sounder and from the ground by Dobson and Brewer spectrophotometers for the locations of Hradec Kralove (Czech Republic, 50 degrees N), Nairobi (Kenya, 1 degrees S) and Springbok (Republic of South Africa, 30 degrees S). The values were used as input parameter to model calculations of erythemally effective irradiance and daily radiant exposure. The differences due to the use of TOC from different sources were analyzed with respect to the Ultraviolet Index (UVI). The UVI was introduced as a tool for sun protection and health care. Therefore, it is of special importance to know the restriction of accuracy. As a tool of health care, the maximum uncertainties are of interest and are described in using the 95%-percentile and the maximum differences. This study shows that differences, i.e. uncertainties (95%-percentile) are in the order of 1 UVI. Independently on the location, however, extreme differences may overstep 3 UVI. For the daily dose the 95% percentile is around 7.5 UVI hours (UVIh) but differences higher than 20 UVIh were also found. PMID- 17115800 TI - Study of singlet oxygen equilibrium in dioctadecyldimethylammonium chloride vesicles employing 2-(n-(N,N,N-trimethylamine)-n-alkyl)-5-alkylfuryl halides. AB - Steady state photolysis and time resolved near infrared luminescence detection were employed to study the reaction kinetics of singlet oxygen with three different lipid-soluble probes incorporated in large unilamellar dioctadecyldimethylammonium chloride (DODAC) vesicles. The probes: 2-(4-(N,N,N trimethylamine)-butyl)-5-dodecylfuryl bromide (DFTA), 2-(12-(N,N,N trimethylamine)-dodecyl)-5-hexylfuryl bromide (HFDA) and 2-(1-(N,N,N trimethylamine)-methyl)-5-methylfuryl iodide (MFMA) are useful in studying both singlet oxygen dynamics and its equilibrium in microcompartmentalized systems because they are actinometers in lipidic microphases. These probes contain a reactive furan ring, which will be located at different depths in the bilayer of DODAC vesicles. In the limit of the approximations, the result indicates an inhomogeneous equilibrium distribution of singlet oxygen across the bilayer. The calculated mean partitioning constant of singlet oxygen equals 2.8 and 8.3 at 20 degrees C and 40 degrees C, respectively, in the order of the previously reported constants for other microorganized systems such as sodium dodecylsulfate and cetyltrimethylammonium halide micelles and water/oil microemulsions. PMID- 17115801 TI - Comparative study of the photophysical behavior of fisetin in homogeneous media and in anionic and cationic reverse micelles media. AB - The 3,3', 4',7 tetrahydroxiflavone (fisetin) is a natural therapeutically active and fluorescent polyhydroxyflavone, with important spectroscopic and biological behavior. Fisetin shows dual emission, with a normal band (N) from the S1 --> S0 transition and the one generated in the excited state (phototautomer; PT) from the intramolecular proton transfer (ESIPT) process. The influence of different interfaces on the ESIPT process of fisetin was investigated in reverse micelles media (RMs) made of the anionic sodium 1,4-bis (2-ethylhexyl) sulfosuccinate (AOT) and cationic benzyl n-hexadecyl dimethylammonium chloride (BHDC) surfactants, in benzene. The studies were carried out by absorption, emission spectroscopy, steady-state anisotropy and time-resolved fluorescence measurements. Fisetin behavior was also investigated in homogeneous media with special emphasis in water and benzene, which are the polar core and the organic pseudofase in the RMs, respectively. In addition, the effect of concentration in benzene and the variation of the pH in water were studied. Fluorescence lifetime measurements show that in water the ESIPT process is independent on the concentration, while in benzene it was possible to detect fluorescent aggregate species (Nas) formed in the ground state. The effect of the pH in water allowed us to identify the anionic fisetin (A-) emission. The studies in RMs show that fisetin interacts specifically with the head of the surfactants, which always results in diminishing the emission of the PT. Also the formation of A- is detected particularly at W0 > 0. Appreciable high anisotropy values are obtained in RMs, as compared with those in fluid homogeneous media, which are independent of the water content confirming that fisetin molecules are anchored in the anionic as well as in the cationic interfaces. PMID- 17115802 TI - Radiometric quantities and units used in photobiology and photochemistry: recommendations of the Commission Internationale de L'Eclairage (International Commission on Illumination). AB - To characterize photobiological and photochemical phenomena, standardized terms and units are required. Without a uniform set of descriptors, much of the scientific value of publications can be lost. Attempting to achieve an international consensus for a common language has always been difficult, but now with truly international scientific publications, it is all the more important. As photobiology and photochemistry both represent the fusion of several scientific disciplines, it is not surprising that the physical terms used to describe exposures and dosimetric concepts can vary from author to author. There are, however, international organizations that were established to minimize the confusion produced by poor or inconsistent technical terminology. This note is to review the standardized terms and provide a background on how such terms are developed, with the hope that all readers will attempt to follow the standardized terminology. PMID- 17115805 TI - Michael Davis: 2006 award for distinguished scientific contributions. AB - Presents the citation for Michael Davis, who received the Award for Distinguished Scientific Contributions "for his major discoveries about the brain circuits underlying basic aspects of behavioral plasticity and learning." A brief profile and a selected bibliography, as well as Davis' award address, entitled Neural Systems Involved in Fear and Anxiety Measured With Fear-Potentiated Startle, accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115806 TI - Neural systems involved in fear and anxiety measured with fear-potentiated startle. AB - A good deal is now known about the neural circuitry involved in how conditioned fear can augment a simple reflex (fear-potentiated startle). This involves visual or auditory as well as shock pathways that project via the thalamus and perirhinal or insular cortex to the basolateral amygdala (BLA). The BLA projects to the central (CeA) and medial (MeA) nuclei of the amygdala, which project indirectly to a particular part of the acoustic startle pathway in the brainstem. N-methyl-D-aspartate (NMDA) receptors, as well as various intracellular cascades in the amygdala, are critical for fear learning, which is then mediated by glutamate acting in the CeA. Less predictable stimuli, such as a long-duration bright light or a fearful context, activate the BLA, which projects to the bed nucleus of the stria terminalis (BNST), which projects to the startle pathway much as the CeA does. The anxiogenic peptide corticotropin-releasing hormone increases startle by acting directly in the BNST. CeA-mediated behaviors may represent stimulus-specific fear, whereas BNST-mediated behaviors are more akin to anxiety. NMDA receptors are also involved in extinction of conditioned fear, and both extinction in rats and exposure-based psychotherapy in humans are facilitated by an NMDA-partial agonist called D-cycloserine. ((c) 2006 APA, all rights reserved). PMID- 17115807 TI - Marcia K. Johnson: 2006 award for distinguished scientific contributions. AB - Presents the citation to Marcia K. Johnson, who received the Award for Distinguished Scientific Contributions "for raising and illuminating fundamental questions about the cognitive and neural processes that constitute the subjective experience of mental life." A brief profile and a selected bibliography, as well as Johnson's award address, entitled Memory and Reality, accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115808 TI - Memory and reality. AB - Although it may be disconcerting to contemplate, true and false memories arise in the same way. Memories are attributions that we make about our mental experiences based on their subjective qualities, our prior knowledge and beliefs, our motives and goals, and the social context. This article describes an approach to studying the nature of these mental experiences and the constructive encoding, revival, and evaluative processes involved (the source monitoring framework). Cognitive behavioral studies using both objective (e.g., recognition, source memory) and subjective (e.g., ratings of memory characteristics) measures and neuroimaging findings are helping to clarify the complex relation between memory and reality. ((c) 2006 APA, all rights reserved). PMID- 17115809 TI - Martin E. P. Seligman: 2006 award for distinguished scientific contributions. AB - Presents the citation for Martin E. P. Seligman, who received the Award for Distinguished Scientific Contributions "for a career spent charging creatively ahead of his field and then pulling his colleagues along." A brief profile and a selected bibliography, as well as the award address, Positive Psychotherapy, accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115810 TI - Positive psychotherapy. AB - Positive psychotherapy (PPT) contrasts with standard interventions for depression by increasing positive emotion, engagement, and meaning rather than directly targeting depressive symptoms. The authors have tested the effects of these interventions in a variety of settings. In informal student and clinical settings, people not uncommonly reported them to be "life-changing." Delivered on the Web, positive psychology exercises relieved depressive symptoms for at least 6 months compared with placebo interventions, the effects of which lasted less than a week. In severe depression, the effects of these Web exercises were particularly striking. This address reports two preliminary studies: In the first, PPT delivered to groups significantly decreased levels of mild-to-moderate depression through 1-year follow-up. In the second, PPT delivered to individuals produced higher remission rates than did treatment as usual and treatment as usual plus medication among outpatients with major depressive disorder. Together, these studies suggest that treatments for depression may usefully be supplemented by exercises that explicitly increase positive emotion, engagement, and meaning. ((c) 2006 APA, all rights reserved). PMID- 17115812 TI - John P. Campbell: award for distinguished scientific applications of psychology. AB - Presents the citation for John P. Campbell who received Award for Distinguished Scientific Applications of Psychology "for his many different contributions to the field of industrial and organizational (I/O) psychology." A brief profile and a selected bibliography accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115814 TI - Mark G. Baxter: award for distinguished scientific early career contributions to psychology. AB - Presents the citation for Mark G. Baxter, who received the Award for Distinguished Scientific Early Career Contributions to Psychology (animal learning and behavior, comparative) "for insightful and incisive contributions in studies of learning, memory, attentional processing, executive function, and goal directed behavior in work that has translated across species." A brief profile and a selected bibliography accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115815 TI - Theodore P. Beauchaine: award for distinguished scientific early career contributions to psychology. AB - Presents the citation for Theodore P. Beauchaine, who received the Award for Distinguished Scientific Early Career Contributions to Psychology (psychopathology) "for core contributions in developmental psychopathology, especially related to the biological underpinnings of various mental disorders among children, sophisticated and elegant quantitative approaches to these issues, and exemplary work on the prevention of such conditions." A brief profile and a selected bibliography accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115816 TI - Angela Bryan: award for distinguished scientific early career contributions to psychology. AB - Presents the citation for Angela Bryan, who received the Award for Distinguished Scientific Early Career Contributions to Psychology (health psychology) "for her outstanding theoretical and applied research on health behavior change." A brief profile and a selected bibliography accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115817 TI - Seth D. Pollak: award for distinguished scientific early career contributions to psychology. AB - Presents the citation for Seth D. Pollak, who received the Award for Distinguished Scientific Early Career Contributions to Psychology (developmental psychology) "for his research on the mechanisms of emotional development using an innovative combination of methods from psychophysics, neuroscience, and behavioral endocrinology." A brief profile and a selected bibliography accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115818 TI - Brian J. Scholl: award for distinguished scientific early career contributions to psychology. AB - Presents the citation for Brian J. Scholl, who received the Award for Distinguished Scientific Early Career Contributions to Psychology "for his brilliantly creative and sweepingly broad research program that addresses the hard questions in cognitive science." A brief profile and a selected bibliography accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115819 TI - Anthony D. Wagner: award for distinguished scientific early career contributions to psychology. AB - Presents the citation for Anthony D. Wagner, who received the Award for Distinguished Scientific Early Career Contributions to Psychology (cognition and human learning) "for outstanding and innovative research on the neural basis of memory and executive control." A brief profile and a selected bibliography accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115822 TI - McCay Vernon: award for distinguished senior career contributions to psychology in the public interest. AB - Presents a citation for McCay Vernon, who received the Award for Distinguished Senior Career Contributions to Psychology in the Public Interest "for fifty-five years of distinguished contributions toward improving the lives of both deaf and deaf-blind individuals." Accompanying the citation are a brief profile and a selected bibliography, as well as Vernon's award address, The APA and Deafness. ((c) 2006 APA, all rights reserved). PMID- 17115823 TI - The APA and deafness. AB - Until the 1960s, people who were Deaf and mentally ill lacked access to psychological treatment. Few mental hospitals and clinics had interpreters available, and few psychologists and mental health professionals had knowledge of sign language. Major court decisions and federal laws have effected change, culminating with the Americans With Disabilities Act of 1990. This legislation gave people who are Deaf the right to equal access to mental health care as well as a host of other opportunities they had been previously denied. New access laws allowed Deaf students to become educated as psychologists, and a number of hearing psychologists who knew sign language entered the field of deafness. These two groups assumed vital roles within the American Psychological Association in addressing the issue of mental health access for people who are Deaf. ((c) 2006 APA, all rights reserved). PMID- 17115824 TI - Rose L. Clark: award for distinguished early career contributions to psychology in the public interest. AB - Presents a citation for Rose L. Clark, who received the Award for Distinguished Early Career Contributions to Psychology in the Public Interest "for her leadership and contributions to the field of psychology in the awareness and advancement of research, practice, and policy on behalf of American Indian/Alaska Native (AI/AN) children, families, and communities." A brief profile and a selected bibliography accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115826 TI - Mark D. Cunningham: award for distinguished contributions to research in public policy. AB - Presents a citation for Mark D. Cunningham, who received the Award for Distinguished Contributions to Research in Public Policy "for his highly distinguished contributions to research on death-sentenced inmates, capital sentencing determinations, and law." Accompanying the citation are a brief profile and a selected bibliography, as well as Cunningham's award address, entitled Dangerousness and Death: A Nexus in Search of Science and Reason. ((c) 2006 APA, all rights reserved). PMID- 17115827 TI - Dangerousness and death: a nexus in search of science and reason. AB - An assertion that a capital offender will engage in future acts of criminal violence is a factor in determining "death worthiness" in many jurisdictions of the United States. The legislative conception and court affirmation of this issue as a capital consideration were products of the parole policy and prison capability of an earlier era as well as of the limitations of risk assessment methodology and findings at that time. The intuitive assumptions that support assertions of future dangerousness as a death penalty issue have been rendered irrelevant by changing prison conditions and parole policies or have been refuted by current findings. This raises important questions for a reasoned public policy in capital sentencing. ((c) 2006 APA, all rights reserved). PMID- 17115830 TI - Janet E. Helms: award for distinguished contributions to education and training in psychology. AB - Presents a citation for Janet E. Helms, who received the Award for Distinguished Contributions to Education and Training in Psychology "for her remarkable contribution to a conceptual and reiterative empirical base that has transformed the study of racism in our time." Accompanying the citation are a brief profile and a selected bibliography, as well as Helms' award address, entitled Fairness Is Not Validity or Cultural Bias in Racial-Group Assessment: A Quantitative Perspective. ((c) 2006 APA, all rights reserved). PMID- 17115831 TI - Fairness is not validity or cultural bias in racial-group assessment: a quantitative perspective. AB - When test scores that differ by racial groups are used for assessment purposes, resulting decisions regarding members of the lower scoring group are potentially unfair. Fairness is defined as the removal from test scores of systematic variance attributable to experiences of racial or cultural socialization, and it is differentiated from test-score validity and cultural bias. Two fairness models for identifying, quantifying, and removing from test scores construct-irrelevant variance attributable to racial or cultural psychological attributes are presented. ((c) 2006 APA, all rights reserved). PMID- 17115832 TI - Michael C. Roberts: award for distinguished contributions to education and training in psychology. AB - Presents a citation for Michael C. Roberts, who received the Award for Distinguished Contributions to Education and Training in Psychology "for his dedication to the education, training, and mentoring of psychologists from undergraduate study through professional career." Accompanying the citation are a brief profile and a selected bibliography, as well as Roberts' award address, entitled Essential Tension: Specialization With Broad and General Training in Psychology. ((c) 2006 APA, all rights reserved). PMID- 17115833 TI - Essential tension: specialization with broad and general training in psychology. AB - The practice fields of psychology develop through specialization in training and education. The recognized specialties play a major role in developing new opportunities for professional psychology and providing quality services for the public. The essential tension comes from the balance of innovation and tradition and, in professional psychology, from the balance of fragmentation and unification. As an example, specialization in clinical child psychology is integrated within the broad and general traditions. The greater degree of focused science and practice in a specialty is the logical consequence of advances of the discipline and profession of psychology. ((c) 2006 APA, all rights reserved). PMID- 17115835 TI - Gregory Daniel Webster: Psi Chi/APA Edwin B. Newman Graduate Research Award. AB - Presents a citation for Gregory Daniel Webster, who received the Psi Chi/APA Edwin B. Newman Graduate Research Award "for an outstanding research paper whose findings report that APA journal articles became shorter in length after the year 2000." A brief profile and a selected bibliography accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115838 TI - Stephen M. Weiss: award for distinguished professional contributions to applied research. AB - Presents the citation for Stephen M. Weiss, who received the Award for Distinguished Professional Contributions to Applied Research "for his work as an internationally recognized leader in the fields of health psychology and behavioral medicine." A brief profile and a selected bibliography accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115840 TI - Jeffrey J. Magnavita: award for distinguished professional contributions to independent or institutional practice in the private sector. AB - Presents the citation for Jeffrey J. Magnavita, who received the Award for Distinguished Professional Contributions to Independent or Institutional Practice in the Private Sector "for his passionate devotion to the advancement of the practice and science of psychotherapy." Accompanying the citation are a brief profile and a selected bibliography, as well as Magnavita's award address, entitled In Search of the Unifying Principles of Psychotherapy: Conceptual, Empirical, and Clinical Convergence. ((c) 2006 APA, all rights reserved). PMID- 17115841 TI - In search of the unifying principles of psychotherapy: conceptual, empirical, and clinical convergence. AB - The search for the principles of unified psychotherapy is an important stage in the advancement of the field. Converging evidence from various streams of clinical science allows the identification of some of the major domains of human functioning, adaptation, and dysfunction. These principles, supported by animal modeling, neuroscience, and developmental psychopathology, along with new theoretical constructs, are demonstrating that human functioning is best conceptualized as holistic and highly interrelated at all domain levels of the total ecological system. Incorporating these findings with evidence accrued over the past century of psychotherapy practice and study allows psychologists to begin to explore the development of a unified system of psychotherapy that is rooted in the clinical sciences. One such attempt to develop a unified framework is presented as a starting point. ((c) 2006 APA, all rights reserved). PMID- 17115843 TI - Thomas J. Fagan: award for distinguished professional contributions to practice in the public sector. AB - Presents the citation for Thomas J. Fagan, who received the Award for Distinguished Professional Contributions to Practice in the Public Sector "for his service as a leader in correctional psychology." A brief profile and a selected bibliography accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115845 TI - Mona M. Amer: APA/APAGS award for distinguished graduate student in professional psychology. AB - Presents the citation of Mona M. Amer, who received the APA/APAGS Award for Distinguished Graduate Student in Professional Psychology "for her outstanding and innovative leadership in addressing the mental health needs of Muslim and Arab Americans." A brief profile and a selected bibliography accompany the citation. ((c) 2006 APA, all rights reserved). PMID- 17115848 TI - Michael Cole: award for distinguished contributions to the international advancement of psychology. AB - Presents the citation for Michael Cole, who received the Award for Distinguished Contributions to the International Advancement of Psychology "for his significant and influential contributions to understanding the cultural and historical construction of human life and the implications of collective social practices for human development." Accompanying the citation are a brief profile and a selected bibliography, as well as Cole's award address, entitled Internationalism in Psychology: We Need It Now More Than Ever. ((c) 2006 APA, all rights reserved). PMID- 17115849 TI - Internationalism in psychology: we need it now more than ever. AB - A concern with international cooperation in psychology has been present since the beginnings of psychology as a science. In this article, the author traces the development of international cooperation from the late 19th century to the present day to document the interesting ways in which the forms of collaboration have always been related to the large social and political contexts of which they are a part. This historical material is followed by an autobiographical account of the author's own involvement in cooperative international research to illustrate how such activity can play out. A few selected collaborative enterprises are then described to emphasize the point that psychology stands to benefit enormously from continued international cooperation, especially in times of international conflict such as that being experienced at the present time. ((c) 2006 APA, all rights reserved). PMID- 17115851 TI - Center for Victims of Torture--Guinea 2004-2005 International Mental Health Team: international humanitarian award. AB - Presents the citation for the Center for Victims of Torture--Guinea 2004-2005 International Mental Health Team, who received the International Humanitarian Award "for their outstanding commitment to the healing of torture victims and to educating the world community about torture and war trauma." A brief profile of the Center is presented, as well as the Center's award address, entitled Trauma Healing in Refugee Camps in Guinea: A Psychosocial Program for Liberian and Sierra Leonean Survivors of Torture and War. ((c) 2006 APA, all rights reserved). PMID- 17115852 TI - Trauma healing in refugee camps in Guinea: a psychosocial program for Liberian and Sierra Leonean survivors of torture and war. AB - From 1999 to 2005, the Minneapolis-based Center for Victims of Torture (CVT) served Liberian and Sierra Leonean survivors of torture and war living in the refugee camps of Guinea. A psychosocial program was developed with 3 main goals: (a) to provide mental health care, (b) to train local refugee counselors, and (c) to raise community awareness about war trauma and mental health. Utilizing paraprofessional counselors under the close, on-site supervision of expatriate clinicians, the treatment model blended elements of Western and indigenous healing. The core component consisted of relationship-based supportive group counseling. Clinical interventions were guided by a 3-stage model of trauma recovery (safety, mourning, reconnection), which was adapted to the realities of the refugee camp setting. Over 4,000 clients were provided with counseling, and an additional 15,000 were provided with other supportive services. Results from follow-up assessments indicated significant reductions in trauma symptoms and increases in measures of daily functioning and social support during and after participation in groups. The treatment model developed in Guinea served as the basis for CVT's ongoing work with survivors in Sierra Leone and Liberia. ((c) 2006 APA, all rights reserved). PMID- 17115853 TI - Global and local processing in adult humans (Homo sapiens), 5-year-old children (Homo sapiens), and adult cotton-top tamarins (Saguinus oedipus). AB - This study compared adults (Homo sapiens), young children (Homo sapiens), and adult tamarins (Saguinus oedipus) while they discriminated global and local properties of stimuli. Subjects were trained to discriminate a circle made of circle elements from a square made of square elements and were tested with circles made of squares and squares made of circles. Adult humans showed a global bias in testing that was unaffected by the density of the elements in the stimuli. Children showed a global bias with dense displays but discriminated by both local and global properties with sparse displays. Adult tamarins' biases matched those of the children. The striking similarity between the perceptual processing of adult monkeys and humans diagnosed with autism and the difference between this and normatively developing human perception is discussed. PMID- 17115854 TI - Comparative analysis of movement characteristics during dead-reckoning-based navigation in humans and rats. AB - Human and rat movement organization was investigated as they searched for randomly located rewards without access to visual information. Under dark conditions, rats foraged for randomly located food pellets (Experiment 1). Blindfolded humans were instructed to search for an ostensible hidden coin using a metal detector (Experiment 2). After locating the food pellet, rats carried it back to the refuge, and after a designated searching time, humans were instructed to return to the start location. Although both species exhibited a high degree of similarity in searching path movement organization and ability to return to the start location, disruption of human searching path organization was associated with impairments in returning to the start location. These results support the vestibular "gain" account of movement organization during dead-reckoning-based navigation. PMID- 17115855 TI - Anticipation of future events in squirrel monkeys (Saimiri sciureus) and rats (Rattus norvegicus): tests of the Bischof-Kohler hypothesis. AB - The Bischof-Kohler hypothesis holds that nonhuman animals cannot anticipate a future event and take appropriate action when that event involves satisfaction of a need not currently experienced. Tests of the Bischof-Kohler hypothesis were performed with squirrel monkeys (Saimiri sciureus) and rats (Rattus norvegicus). In experimental trials with both species, a nonthirsty animal had its water bottle removed and then chose between a smaller and larger quantity of food. Consumption of the food induced thirst. Choice of the smaller quantity led to the return of the water bottle sooner than choice of the larger quantity. Monkeys reversed their baseline preference for the larger quantity of food when the experimental contingencies were introduced, but rats continued to prefer the larger amount. Although the rat findings support the Bischof-Kohler hypothesis, the monkey findings challenge it. PMID- 17115856 TI - Performance of pigeons (Columba livia) on maze problems presented on the LCD screen: in search for preplanning ability in an avian species. AB - The authors examined how pigeons (Columba livia) perform on 2-dimensional maze tasks on the LCD monitor and whether the pigeons preplan the solution before starting to solve the maze. After training 4 pigeons to move a red square (the target) to a blue square (the goal) by pecking, the authors exposed them to a variety of detour tasks having lines as a barrier. A preview phase was introduced, during which the pigeons were not allowed to peck at the monitor. Results of a set of experiments suggest that our pigeons successfully learned to solve these tasks, that they came to take an efficient strategy as the barriers became complex, and that they possibly preplan its solution, at least on familiar, well-practiced tasks. PMID- 17115857 TI - Decision making and interference in the domestic cat (Felis catus). AB - Domestic cats (Felis catus) received a task in which an obstacle was introduced at the time they were ready to reach for 1 among 2 or 3 baited targets. The results revealed that when the initially chosen direction (i.e., before the introduction of the obstacle) and the new one were orthogonal, the cats reassessed distance, angular deviation, and visibility once they had turned around the obstacle (Experiments 1, 2, and 3). Then the cats chose a different target than the one that would have been chosen had there been no obstacle. When both the initially chosen and the new direction matched (Experiment 4), the cats kept intact their initial decision. The discussion focuses on the nature of the representational system of the cat. The discussion also emphasizes comparisons with dogs (i.e., cooperative hunters). PMID- 17115858 TI - The structure of individual differences in batteries of rapid acquisition tasks in mice. AB - Two experiments examined the structure of individual differences in mice by means of tasks that produced significant acquisition within 1 session. In Experiment 1, 5 cognitive tasks-detour, winshift, olfactory discrimination, fear conditioning, and operant acquisition-were used in conjunction with two control procedures: an open field and a light- dark test. In Experiment 2, some modifications were made to the tasks used in the 1st experiment, and 3 new tasks were used in conjunction with the same control procedures. The battery consisted of 5 learning tasks: detour, Hebb-Williams, radial maze, olfactory foraging, and fear conditioning. Results of both experiments indicate that when cognitive tasks and control procedures were included in principal-components analyses most of the variance attached principally to individual tasks rather than to a general component as is found typically in human cognitive batteries. When control procedures were eliminated, there was better evidence for the presence of a general cognitive factor, particularly in Experiment 2. PMID- 17115859 TI - Chimpanzees (Pan troglodytes) remember the location of a hidden food item after altering their orientation to a spatial array. AB - Two chimpanzees (Pan troglodytes) had a direct view of an experimenter placing a food item beneath one of several cups within a horizontal spatial array. The chimpanzees then were required to move around the spatial array, shifting their orientation to the array by 180 degrees . Both chimpanzees remembered the location of the food item. In the next experiment, a visual barrier was placed between the chimpanzees and the spatial array after the food item had been hidden to prevent visual tracking of the location of the object during the chimpanzees' movement. One chimpanzee remembered the location of the hidden item in this variation. These results demonstrate another capacity for spatial memory in this species that complements data indicating chimpanzee spatial memory for invisible displacements, array rotations, and array transpositions. PMID- 17115860 TI - Dominance interactions in young adult paper wasp (Polistes dominulus) foundresses: a playlike behavior? AB - The idea that insects play has often aroused skepticism. Nevertheless, the authors investigated the occurrence of a playlike behavior in young individuals of a paper wasp. Polistes dominulus foundresses hibernate in aggregations and found associative nests in the spring. In aggregations, wasps exhibit most of the rank-dependent behaviors that they will use in the nest 6 months later. In aggregations, precocious dominance interactions are performed with exaggerated frequencies, whereas aggression and trophallaxis are rare. Dominance behavior in aggregations is apparently useless, but it probably allows wasps to assess their dominance potential without any apparent reproductive competition. Surprisingly, these interactions may be best interpreted as play because dominance behavior in aggregations shares several features with mammalian play fighting. PMID- 17115861 TI - One-sided limb preference is linked to alternating-limb locomotion in anuran amphibians. AB - Amphibians provide a unique opportunity for identifying possible links between lateralized behaviors, locomotion, and phylogeny and for addressing the origin of lateralized behaviors of higher vertebrates. Five anuran species with different locomotive habits were tested for forelimb and hind limb preferences during 2 stereotyped behavior sequences--wiping a foreign object off their snout and righting themselves from the overturned position. The experiments were analyzed in a broader context of previous findings on anuran lateralization involving 11 anuran species that were studied within the same experimental paradigms. This analysis shows that one-sided forelimb and hind limb motor lateralization in anurans is strongly associated with alternating-limb locomotion and other unilateral limb activity. Conclusions reached for anuran amphibians may be applicable to other vertebrates possessing paired appendages-the degree of lateralization in motor response depends on the mode of locomotion used by a species. PMID- 17115862 TI - Nonselective maternal bonding but pup recognition in the subterranean rodent Ctenomys pearsoni. AB - This study characterizes the maternal behavior in the subterranean rodent Ctenomys pearsoni and determines whether lactating females develop a selective bonding to their own pups in addition to maternal responsiveness. Mother-litter interactions were observed daily for 2 months; during the 1st postpartum week, selectivity and preference tests for own or alien pups were performed. Lactating females exhibited all patterns of maternal behavior observed in other rodent species and equally responded with care-taking behaviors to their own and alien pups, although they were able to discriminate between them, showing preference for their own pups. Results are discussed, taking into account the possible role of this behavior in shaping different aspects of life underground in ctenomyid species. PMID- 17115863 TI - How capuchin monkeys (Cebus apella) quantify objects and substances. AB - Humans and nonhuman animals appear to share a capacity for nonverbal quantity representations. But what are the limits of these abilities? Results of previous research with human infants suggest that the ontological status of an entity as an object or a substance affects infants' ability to quantify it. We ask whether the same is true for another primate species-the New World monkey Cebus apella. We tested capuchin monkeys' ability to select the greater of two quantities of either discrete objects or a nonsolid substance. Participants performed above chance with both objects (Experiment 1) and substances (Experiment 2); in both cases, the observed performance was ratio dependent. This finding suggests that capuchins quantify objects and substances similarly and do so via analog magnitude representations. PMID- 17115864 TI - Sexual fetishism in a quail (Coturnix japonica) model system: test of reproductive success. AB - In the present study, the authors explored the reproductive consequences of fetishistic behavior in a previously developed animal model of sexual fetishism (F. Koksal et al., 2004). Male domesticated quail (Coturnix japonica) received sexual conditioning trials in which a terrycloth object (the conditioned stimulus [CS]) was paired with the opportunity to copulate with a female quail (the unconditioned stimulus). Approximately half of the male quail came to copulate with the CS object and were considered to have developed fetishistic behavior. Each of the male quail was then tested with a female quail, whose eggs were incubated to determine rates of fertilization. The CS object was present for 30 s before and during the copulation test. Fetishistic male quail were slower to achieve cloacal contact with the female quail and showed less efficient copulatory behavior. However, they fertilized a greater proportion of eggs than nonfetishistic male quail. These results are unexpected from previous studies of the relationship between reproductive success and copulatory behavior and are discussed in terms of how fetishistic behavior directed toward an inanimate object may modify male-female interactions. PMID- 17115865 TI - Effects of enclosure size on sexual behavior of Japanese quail (Coturnix japonica). AB - The authors determined whether results of experiments on copulatory and affiliative behavior of pairs of Japanese quail (Coturnix japonica) conducted in a closely confining apparatus would predict behavior in a large enclosure in which female quail could avoid contact with male quail. As found previously in studies of closely confined quail, in a large enclosure containing numerous barriers, both unmated female quail and mated female quail laying unfertilized eggs were more likely to remain near a confined male quail than were mated female quail laying fertilized eggs. Furthermore, the number of copulations that a pair engaged in when closely confined predicted the number of copulations that they engaged in when they were in the large enclosure. Patterns of affiliation and of mating in a confining laboratory apparatus thus predicted behavior in a larger enclosure that provided female quail with opportunity to avoid contact with male quail. PMID- 17115866 TI - Cone excitation ratios correlate with color discrimination performance in the horse (Equus caballus). AB - Six horses (Equus caballus) were trained to discriminate color from grays in a counterbalanced sequence in which lightness cues were irrelevant. Subsequently, the pretrained colors were presented in a different sequence. Two sets of novel colors paired with novel grays were also tested. Performance was just as good in these transfer tests. Once the horse had learned to select the chromatic from the achromatic stimulus, regardless of the specific color, they were immediately able to apply this rule to novel stimuli. In terms of the underlying visual mechanisms, the present study showed for the first time that the spectral sensitivity of horse cone photopigments, measured as cone excitation ratios, was correlated with color discrimination performance, measured as accuracy, repeated errors, and latency of approach. PMID- 17115867 TI - Deterioration of planning ability with age in Japanese monkeys (Macaca fuscata). AB - To investigate the ability of aged monkeys to plan and the effect of aging on this ability, performance in a food retrieval task was assessed in aged and younger Japanese monkeys (Macaca fuscata). In this task, the monkeys had to retrieve food items by selecting from a set of 9 holes, each of which contained 1 food item. Results showed that task performance declined significantly with age. All monkeys showed, to a greater or lesser extent, some consistent patterns in their sequence of selecting holes for retrieving the food item. An analysis of these selection patterns indicated that the younger monkeys showed more consistent sequences in selection than the aged monkeys. Furthermore, success in the task performance correlated strongly with higher consistency in the sequence of selecting holes. The authors simulated performance for this task by monkeys without any strategies or plans. The results suggest that the empirical data were far more systematic than the simulated data. Thus, the authors conclude that Japanese monkeys have the ability to plan and that this ability to plan deteriorates with age. PMID- 17115868 TI - Aripiprazole as a potential pharmacotherapy for stimulant dependence: human laboratory studies with d-amphetamine. AB - Amphetamine and cocaine dependence present significant public health concerns, yet no broadly effective pharmacotherapy for stimulant dependence has been developed. Two human laboratory studies are reviewed that tested the ability of aripiprazole, a novel antipsychotic with partial agonist activity at D2 dopamine receptors, to alter the behavioral effects of stimulants using d-amphetamine as a model agent. In each of these experiments, volunteers learned to discriminate 15 mg d-amphetamine (i.e., > or =80% drug-appropriate responding over 4 consecutive sessions). The effects of a range of doses of d-amphetamine (0, 2.5, 5, 10, and 15 mg) were then tested alone and following pretreatment with aripiprazole (20 mg in Experiment 1; 10 mg in Experiment 2). In Experiment 1, aripiprazole (20 mg) attenuated the discriminative stimulus and many of the subject-rated effects of amphetamine. Aripiprazole alone produced performance decrements. To determine whether a lower dose of aripiprazole would also attenuate the behavioral effects of d-amphetamine without impairing performance, Experiment 2 was conducted. Aripiprazole (10 mg) failed to alter the discriminative-stimulus effects but attenuated some of the subject-rated effects of d-amphetamine. This dose of aripiprazole did not impair performance. The results of these experiments indicate that aripiprazole may have clinical utility in treating stimulant dependence. Future human laboratory research should better model the clinical use of aripiprazole by examining the effects of chronic aripiprazole combined with either methamphetamine or cocaine in dependent individuals. A large-scale clinical trial is also needed to evaluate the efficacy of aripiprazole for the treatment of stimulant dependence. PMID- 17115869 TI - Altered regional blood volume in chronic cannabis smokers. AB - The quantitative measurement of cerebral perfusion is crucial for the study of both normal and impaired human brain function. Although cannabis is the most commonly abused illicit substance in the United States, its effects on cerebral blood volume (CBV) have not been fully examined. The objective of the present study was to examine differences in relative regional blood volume in focal regions of interest--including the frontal lobe, the temporal lobe, and the cerebellum--during a period of supervised abstinence from cannabis. Dynamic susceptibility contrast MRI data were collected on 12 current, long-term daily cannabis users between 6 and 36 hr after the subjects' last reported cannabis use. Resting-state CBV images were also acquired in 17 healthy comparison subjects. Data were acquired in the axial plane with a 1.5-Tesla GE Signa scanner following a bolus of gadolinium contrast agent. Cannabis users demonstrated significantly increased blood volumes in the right frontal area (p < .05), in the left temporal area (p < .005), and in the cerebellum (p < .005) relative to comparison subjects. Among the cannabis users, there were no significant correlations between regional blood volumes and either total lifetime episodes of smoking or urinary tetrahydrocannabinol concentrations. These findings have important implications for understanding the effects of chronic heavy cannabis use on brain function. It would be of interest to extend the investigation beyond 6-36 hr of abstinence from cannabis to determine whether increased CBV values persist for several weeks or eventually normalize. PMID- 17115870 TI - A preliminary investigation of rapid smoking as a lapse-responsive treatment for tobacco dependence. AB - Lapses within the first 2 weeks of a smoking cessation attempt are strongly associated with a return to regular smoking (S. L. Kenford et al., 1994). Unfortunately, little is known about how to prevent an initial lapse from progressing to a full relapse, and presently there are no validated lapse responsive therapeutic interventions. The present study tested the efficacy and feasibility of rapid smoking plus counseling as a novel lapse-responsive intervention. Sixty-seven participants enrolled in a smoking treatment program involving brief counseling and a 9-week course of bupropion. Beginning on the quit day, participants' smoking behavior was tracked daily for 14 days. Once an early smoking lapse was identified, participants were randomly assigned to receive either 3 sessions of rapid smoking plus counseling or no intervention (usual care). Consistent with previous research, participants who smoked during the first 2 weeks of the quit attempt had significantly poorer 6-month outcomes (3% abstinent) than did those who did not smoke (64% abstinent). Compared with early abstainers, early lapsers were more nicotine dependent and reported greater cravings and lower confidence in their ability to abstain from smoking during the first 48 hours of abstinence. As expected, rapid smoking produced a variety of aversive effects, including increased nausea, dizziness, and vomiting as well as sharply decreased cravings to smoke. However, rapid smoking did not improve abstinence outcomes relative to usual care. Although rapid smoking has been shown to be an effective treatment for initial smoking cessation, in this preliminary study the authors failed to demonstrate its effectiveness as a lapse-responsive treatment. PMID- 17115871 TI - Cognitive and subjective acute dose effects of intramuscular ketamine in healthy adults. AB - Ketamine is a noncompetitive N-methyl-D-aspartate (NMDA) antagonist. Given the purported role of the NMDA receptor in long-term potentiation, the primary purpose of the present study was to further understand the dose-related effects of ketamine on memory. The study was also designed to provide information about the relative effects of ketamine on memory versus nonmemory effects and to more fully characterize ketamine's overall pattern and time course of effects. Single intramuscular injections of ketamine (0.2 mg/kg, 0.4 mg/kg) were administered to 18 healthy adult volunteers using a double-blind, placebo-controlled, crossover design. Word lists were used to evaluate episodic memory (free recall, recognition memory, source memory) and metamemory. Working memory, time estimation, psychomotor performance, and subjective effects were assessed repeatedly for 5 hours after drug administration. Ketamine selectively impaired encoding (as measured by free recall) while sparing retrieval, working memory while sparing attention, and digit symbol substitution task speed while sparing accuracy. Ketamine did not significantly impair recognition or source memory, metamemory, or time estimation. There were no hallucinations or increases in mystical experiences with ketamine. Memory measures were less sensitive to ketamine effects than subjective or psychomotor measures. Subjective effects lasted longer than memory and most psychomotor impairments. Ketamine produces selective, transient, dose- and time-related effects. In conjunction with previous studies of drugs with different mechanisms of actions, the observed selectivity of effects enhances the understanding of the pharmacological mechanisms underlying memory, attention, psychomotor performance, and subjective experience. PMID- 17115872 TI - Clubgoers and their trendy cocktails: implications of mixing caffeine into alcohol on information processing and subjective reports of intoxication. AB - Alcoholic drink preferences in college students have made an interesting shift recently, with trends in consumption leaning toward caffeinated alcohol in various forms (e.g., Red Bull and vodka or caffeinated beers such as Anheuser Busch's B-to-the-E). Despite the dramatic rise in popularity of these beverages, little research has examined the combined effects of alcohol and caffeine, which is problematic for adequately informing the public about the risk or lack thereof of these drinks. The purpose of this study was to directly investigate the acute effects of alcohol and caffeine, alone and in combination, on well-validated measures of cognitive performance and subjective intoxication in social drinkers. Participants (N = 12) performed a psychological refractory period task that measured dual-task interference as the prolonged reaction time to complete the 2nd of 2 tasks performed in close temporal sequence. Performance was tested under 2 active doses and 1 placebo dose of caffeine (0.0 mg/kg, 2.0 mg/kg, and 4.0 mg/kg) in combination with 1 active dose and 1 placebo dose of alcohol (0.0 g/kg and 0.65 g/kg). As expected, alcohol impaired task performance by increasing dual task interference and increasing errors. The coadministration of caffeine counteracted the effects of alcohol on interference but had no effect on the degree to which alcohol increased errors. Subjective measures of intoxication showed that coadministration of caffeine with alcohol reduced participants' perceptions of alcohol intoxication compared with administration of alcohol alone. The results highlight the complexity of drug interactions between alcohol and caffeine. PMID- 17115873 TI - Menstrual disturbance and galactorrhea in people taking conventional antipsychotic medications. AB - Endocrine disturbances are emerging as major side effects of antipsychotic medications. Of particular note is the profile of menstrual disturbance and galactorrhea as a consequence of hyperprolactinemia (A. Weick & P. M. Haddad, 2003), a sequela of antidopaminergic action at the hypothalamopituitary axis. Research into the clinical aspects of this sensitive issue is sparse. The authors completed a cross-sectional descriptive study of 50 patients on conventional antipsychotic medications. The prevalence of menstrual disturbance was 54%, and the prevalence of amenorrhea was 12%. Symptoms of galactorrhea were present in 32% of patients. A history of pregnancy and childbirth was noted to be significantly associated with the development of galactorrhea (p = .01). The authors hypothesized that pregnancy and lactation might sensitize the hypothalamopituitary axis for further development of hyperprolactinemia due to medications. PMID- 17115874 TI - Alcohol and erectile response: the effects of high dosage in the context of demands to maximize sexual arousal. AB - Although drinking often precedes men's sexual activity, basic questions about alcohol's effects on men's sexual arousal remain unanswered. Inconsistencies in findings from studies examining subjective and physiological effects on erectile functioning suggest these effects are context specific, for example, dependent on whether a man wants to maximize or suppress his arousal. To address unresolved questions about alcohol and erectile functioning, the authors evaluated the effects of high blood alcohol concentrations (BACs) and arousal instructional demands on indices of penile circumference change and self-reported sexual arousal. In Study 1, a target BAC of .10% (vs. .00%) attenuated peak circumference change from a neutral baseline but did not affect mean change, latency to arousal onset (a 5% increase in circumference from baseline), latency to peak achieved arousal, or subjective arousal, which correlated moderately with physiological indices. In Study 2, instructions to maximize (vs. suppress) arousal increased peak and mean circumference change and interacted with a target BAC of .08% (vs. .00%) to influence latency to arousal onset. Sober men instructed to maximize showed a shorter latency to arousal onset than did those instructed to suppress arousal; however, intoxicated men did not show a differential pattern. Moreover, compared with intoxicated counterparts, sober men instructed to maximize arousal showed a marginally shorter latency to arousal onset. Overall, alcohol and arousal instructions had small but discernible effects. Findings highlight the importance of contextual factors in alcohol's impact on erectile functioning. PMID- 17115875 TI - Spontaneous recovery and dishabituation of ethanol-reinforced responding in alcohol-preferring rats. AB - This study examined whether habituation, a decrease in responsiveness to a repeatedly presented stimulus, occurs to ethanol reinforcers in alcohol preferring (P) rats. Three fundamental properties of habituation were evaluated: generality, spontaneous recovery, and dishabituation. In each experiment, P rats' lever pressing was reinforced by 10% ethanol on a variable-interval 15-s schedule during 50-min sessions. Experiment 1 evaluated the generality of habituation to repeatedly presented stimuli by using ethanol and water reinforcers. Rates of responding were higher for ethanol than they were for water. Additionally, the within-session patterns of responding differed for each reinforcer, suggesting that the pattern of responding was specific to the exact nature of the repeatedly presented reinforcer. Experiment 2 examined spontaneous recovery, an increase in responsiveness to a habituated stimulus when that stimulus is not presented for a time, by separating experimental sessions by 5 min, 2 hr, or 24 hr. Early-session rates of responding during Session 2 were slower than the corresponding rates during Session 1 when sessions were separated by 5 min or 2 hr. Response rates and within-session patterns of responding during Sessions 1 and 2 were similar when sessions were separated by 24 hr. Experiment 3 tested for dishabituation, a restoration of responsiveness following the presentation of an extraneous stimulus, by presenting a tone or a light 24 min and 55 s into the session. Rates of responding temporarily increased after the tone was presented. The results of these experiments support the idea that habituation contributes to the regulation of ethanol consumption. PMID- 17115876 TI - Fluvoxamine effects on concurrent ethanol- and food-maintained behaviors. AB - In previous studies, the selective serotonin reuptake inhibitor fluvoxamine preferentially reduced responding for ethanol compared with responding for food under conditions in which each was available alone in separate groups or in the same subjects under a multiple schedule in which baseline response rates were matched. The impact of providing concurrent access to food on pharmacological effects on ethanol self-administration remains largely unexplored. In this study, acute doses of fluvoxamine (3.0-17.8 mg/kg) were administered 30 min before the experimental session to Lewis rats responding under a concurrent fixed-ratio, fixed-ratio schedule of ethanol and food presentation. Ratios for food were adjusted for each subject to provide matched rates of food and ethanol reinforcement across the 30-min session. Although the number of ethanol and food deliveries did not significantly differ under baseline conditions, response rates did differ. Following fluvoxamine administration, responding for food was decreased more than responding for ethanol. This differential effect did not appear to be related to response rate or fixed-ratio size. Thus, the selectivity of fluvoxamine on ethanol- versus food-maintained responding depends on the context in which the behavior occurs. Such results may help explain inconsistencies between preclinical results and those in humans, and could provide insight into the behavioral determinants of pharmacological effects on ethanol self-administration. PMID- 17115877 TI - Essential tips for measuring levels of consumer satisfaction with rural health service quality. AB - INTRODUCTION: Quality of health services is a matter of increasing importance to health authorities. Monitoring consumer satisfaction of health care is an important input to improving the quality of health services. This article highlights a number of important considerations learned from rural consumer studies relevant to ensuring the valid measurement of consumer satisfaction with rural health services, as a means of contributing to quality improvements. METHODS: This article compares two methods of analysing rural consumers' satisfaction with healthcare services. In one study of three rural communities in western New South Wales (NSW) and eight communities in north-west Victoria, residents were asked to rate their satisfaction with five key aspects of local health services (availability, geographical accessibility, choice, continuity, economic accessibility as measured by affordability) using a 5 point Likert scale from: one = very satisfied to five = very dissatisfied. An alternative method of assessing levels of consumer satisfaction was undertaken in the survey of eight rural communities in north-west Victoria by investigating consumers' experiences with actual and potential complaints in relation to health services. RESULTS: Both the NSW and Victorian respondents reported generally high levels of satisfaction with the five indicators of quality of health care. At the same time, 11% of Victorian study respondents reported having made a complaint about a health service in the previous 12 months, and one-third of the Victorian respondents reported experiences with their health services about which they wanted to complain but did not, over the same period. CONCLUSIONS: Interpretation of apparent consumer satisfaction with their health services must take particular account of the measures and research methods used. In assessing consumer satisfaction with health services in rural areas, specific attention should be given to maximising the engagement of rural consumers in order to ensure representativeness of findings, and to minimise possible biases in satisfaction ratings associated with the use of particular tools. PMID- 17115878 TI - Transition of patients from chronic kidney disease to end-stage renal disease: better practices for better outcomes. PMID- 17115879 TI - Notes from the field: the economic value chain in disease management organizations. AB - The disease management (DM) "value chain" is composed of a linear series of steps that include operational milestones in the development of knowledge, each stage evolving from the preceding one. As an adaptation of Michael Porter's "value chain" model, the process flow in DM moves along the following path: (1) data/information technology, (2) information generation, (3) analysis, (4) assessment/recommendations, (5) actionable customer plan, and (6) program assessment/reassessment. Each of these stages is managed as a major line of product operations within a DM company or health plan. Metrics around each of the key production variables create benchmark milestones, ongoing management insight into program effectiveness, and potential drivers for activity-based cost accounting pricing models. The value chain process must remain robust from early entry of data and information into the system, through the final presentation and recommendations for our clients if the program is to be effective. For individuals involved in the evaluation or review of DM programs, this framework is an excellent method to visualize the key components and sequence in the process. The value chain model is an excellent way to establish the value of a formal DM program and to create a consultancy relationship with a client involved in purchasing these complex services. PMID- 17115880 TI - A randomized trial of primary intensive care to reduce hospital admissions in patients with high utilization of inpatient services. AB - Randomized controlled trials of case management in primary care have been infrequent and contradictory. The aim of this study was to determine if a clinic based ambulatory case management intervention, Primary Intensive Care (PIC), would reduce hospital utilization and total cost and/or improve health outcomes among primary care patients with a recent history of high use of inpatient services. Current patients with > or =2 hospital admissions per year in the 12-18 months prior to recruitment in an urban primary care clinic were enrolled in a randomized clinical trial. Patients were randomized to the PIC intervention or usual care. PIC patients underwent a comprehensive multidisciplinary assessment with the result being a team-generated plan. The PIC team nurse practitioner served as case manager for the 12 months of follow-up and provided services designed to implement the care plan for those in the experimental group. Health care use, function, and a medication adherence scale were measured at baseline and at 12 months. There were no significant differences when either comparing the number of admissions pre and post enrollment within groups or the followup results post intervention between groups. A similar result was noted for the number of emergency department visits. The number of clinic visits increased in the intervention group by 1.5 visits per year which was statistically significant when compared to the control group. Overall functional status, health outcomes, and the Mental Health Functional Status subscore did not change significantly in either group during the study. We were unable to detect a difference in hospital use or functional status, mental health function, or medication adherence among patients who require frequent hospital admissions using our intervention. PMID- 17115881 TI - Cost of caring for Medicare beneficiaries with Parkinson's disease: impact of the CMS-HCC risk-adjustment model. AB - Previous studies have demonstrated that Medicare risk-adjusted capitation models do not adequately compensate programs serving primarily disabled or frail populations. Using the Medicare Current Beneficiary Survey, we demonstrate that the Centers for Medicare and Medicaid Services-Hierarchical Condition Categories (CMS-HCC) model calculates Medicare capitation payments for Parkinson's patients more accurately than for the general population. The discrepancies between the predicted and actual expenditures estimated at various disability levels were smaller for Parkinson's patients than for other beneficiaries. If the CMS-HCC payment model were to apply to programs that draw a significant percentage of their participants from the Parkinson's disease community, these programs likely would be compensated fairly. PMID- 17115882 TI - Innovative reflecting interview: effect on high-utilizing patients with medically unexplained symptoms. AB - This pilot study was conducted to determine the effect of an innovative reflecting interview on the health care utilization, physical health, mental function, and health care satisfaction of high-utilizing primary care patients with medically unexplained physical symptoms. Twenty-four high-utilizing patients met study selection criteria and were randomly assigned to a no-intervention control group or a reflecting interview intervention group. Outcomes were measured at 4 weeks, 6 months, and 1 year after the date of study enrollment. Results indicated that high-utilizing patients with medically unexplained physical symptoms who participated in a reflecting interview had reduced total health care costs, primarily through the reduction of hospitalization or inpatient expenses, despite a modest increase in outpatient primary care clinic visits. These data suggest that participation in a reflecting interview and regular visits with a primary care clinician can decrease health care utilization without adversely affecting patient satisfaction. PMID- 17115883 TI - Improvement of LDL-C laboratory values achieved by participation in a cardiac or diabetes disease management program. AB - Poor lipid control is a risk factor for cardiovascular diseases and diabetes complications. Frequently, however, patients with these diseases do not achieve blood lipid levels recommended by current standards of care. A retrospective study of 67,244 members eligible for disease management (DM) was initiated to evaluate the ability of interventions to promote improvement in low-density lipoprotein cholesterol (LDL-C) laboratory values for people with cardiovascular diseases or diabetes. The baseline trend in improving LDL-C values in the absence of DM was established. A two-year period prior to the start of the DM intervention was examined to measure the mean percent change in LDL-C values that was occurring in the population. The mean percent change observed for this pre intervention group was then compared to the change in LDL-C values observed during the DM study period. A significant reduction in elevated LDL-C values (F test; p < 0.0001) was observed for members who participated in the DM interventions, even when elevated LDL-C was defined as low as > or =70 mg/dL. Members with LDL-C values within threshold limits maintained these levels during the DM program. The significant reduction in elevated LDL-C values and maintenance of optimal values (< 100 mg/dL) was observed over the course of 3 years of participation in a DM program. A subset of the population also was examined to assess the impact of telephone intervention on reducing elevated LDL C values. A significant relationship between receiving care calls and reduction in elevated LDL-C levels was observed; members who received calls achieved up to a 32.5% relative reduction in elevated LDL-C values compared to members who did not receive calls. In conclusion, these findings demonstrate the ability of DM interventions to assist a large, geographically diverse member population in reducing a clinical laboratory value. PMID- 17115884 TI - DMAA: defining quality in health care coordination. PMID- 17115885 TI - Protein tyrosine phosphorylation and reversible oxidation: two cross-talking posttranslation modifications. AB - In addition to protein phosphorylation, redox-dependent posttranslational modification of proteins is emerging as a key signaling system, conserved throughout evolution, and influencing many aspects of cellular homeostasis. Recent data have provided new insight about the interplay between phosphorylation and redox-dependent signaling, and reactive oxygen species have been included among intracellular signal transducers of growth factor and extracellular matrix receptors. Both tyrosine phosphorylation and thiol oxidation are reversible and dynamic, and this review will particularly focus on the cross-talk between these posttranslational protein regulatory means. Although these modifications share their reversibility, their effects on enzymatic activity of protein tyrosine phosphatases (PTPs) and protein tyrosine kinases (PTKs) may be even opposite. Indeed, while tyrosine phosphorylation is frequently correlated to enzyme activation, thiol oxidation leads to inactivation of PTPs and to superactivation of PTKs. Several papers describe that both these modifications occur during the same input, (i.e., cell proliferation and motility induced by numerous growth factors and cytokines). The review will discuss several aspects of phosphorylation?oxidation interplay, describing both convergent and divergent features of the integrated and coordinated function of PTPs and PTKs during signaling. PMID- 17115886 TI - Thioredoxin and related molecules--from biology to health and disease. AB - Thioredoxin and glutaredoxin systems in mammalian cells utilize thiol and selenol groups to maintain a reducing intracellular redox state acting as antioxidants and reducing agents in redox signaling with oxidizing reactive oxygen species. During the last decade, the functional roles of thioredoxin in particular have continued to expand, also including novel functions such as a secreted growth factor or a chemokine for immune cells. The role of thioredoxin and glutaredoxin in antioxidant defense and the role of thioredoxin in controlling recruitment of inflammatory cells offer potential use in clinical therapy. The fundamental differences between bacterial and mammalian thioredoxin reductases offer new principles for treatment of infections. Clinical drugs already in use target the active site selenol in thioredoxin reductases, inducing cell death in tumor cells. Thioredoxin and binding proteins (ASK1 and TBP2) appear to control apoptosis or metabolic states such as carbohydrate and lipid metabolism related to diseases such as diabetes and atherosclerosis. PMID- 17115887 TI - Mechanisms of cell death in oxidative stress. AB - Reactive oxygen or nitrogen species (ROS/RNS) generated endogenously or in response to environmental stress have long been implicated in tissue injury in the context of a variety of disease states. ROS/RNS can cause cell death by nonphysiological (necrotic) or regulated pathways (apoptotic). The mechanisms by which ROS/RNS cause or regulate apoptosis typically include receptor activation, caspase activation, Bcl-2 family proteins, and mitochondrial dysfunction. Various protein kinase activities, including mitogen-activated protein kinases, protein kinases-B/C, inhibitor-of-I-kappaB kinases, and their corresponding phosphatases modulate the apoptotic program depending on cellular context. Recently, lipid derived mediators have emerged as potential intermediates in the apoptosis pathway triggered by oxidants. Cell death mechanisms have been studied across a broad spectrum of models of oxidative stress, including H2O2, nitric oxide and derivatives, endotoxin-induced inflammation, photodynamic therapy, ultraviolet-A and ionizing radiations, and cigarette smoke. Additionally ROS generated in the lung and other organs as the result of high oxygen therapy or ischemia/reperfusion can stimulate cell death pathways associated with tissue damage. Cells have evolved numerous survival pathways to counter proapoptotic stimuli, which include activation of stress-related protein responses. Among these, the heme oxygenase-1/carbon monoxide system has emerged as a major intracellular antiapoptotic mechanism. PMID- 17115888 TI - Hypertension caused by transgenic overexpression of Rac1. AB - Reactive oxygen species, including superoxide, are important mediators of the pathophysiology of hypertension. In the vasculature, superoxide antagonizes nitric oxide (NO*), resulting in increased vascular tone. The GTP binding protein Rac regulates a wide variety of cellular functions, including the activation of NADPH oxidase, the major source of O2*-in the blood vessel wall. An hypothesis is that Rac1 may act as an important regulator of vascular O2*- production, contributing to the balance between O2*- and NO* and maintaining consequent homeostasis of blood pressure. To alter the activity of vascular NADPH oxidase, the authors developed a transgenic animal model that overexpresses the human cDNA of the constitutively active mutant of Rac1 (RacCA) in smooth muscle cells using the smooth muscle +/--actin promoter. The RacCA transgenic had excessive amounts of O2*- in the vessel wall that, which led to heightened production of peroxynitrite, as detected by increased protein nitration and reduced NO* levels. RacCA mice developed moderate hypertension, which was corrected by N-acetyl-L cysteine (NAC). RacCA transgenic mice also developed left ventricular hypertrophy as a secondary effect of pressure overload. The data suggest that Rac1 is a critical regulator of the redox state of blood vessels and homeostasis of blood pressure. PMID- 17115889 TI - The red wine antioxidant resveratrol prevents cardiomyocyte injury following ischemia-reperfusion via multiple sites and mechanisms. AB - The objective was a comprehensive investigation of the mechanisms and sites of resveratrol cardioprotection during and following ischemia-reperfusion (I-R) injury, and to determine whether direct preservation of cardiomyocytes is an important site of cardioprotection. We now provide the first definitive evidence that resveratrol specifically protects cardiomyocytes from I-R injury via a combination of suppression of superoxide levels and activation of potassium channels. This protection is apparent whether resveratrol is present for the full duration of the insult or only on recovery. In addition, resveratrol improved postischemic recovery of left ventricular contractile function, attenuated myocardial injury, and increased myocardial activation of the survival kinase Akt in the intact heart. Furthermore, resveratrol elicited direct concentration dependent protective actions on the vasculature (vasorelaxation, superoxide suppression) and enhanced endothelium-dependent vasodilatation. Resveratrol thus targets a number of consequences of myocardial I-R, including release of reactive oxygen species, loss of recovery of contractile function, and impaired endothelium-dependent vasodilatation. Previous evidence indicates that resveratrol elicits potent preconditioning in the heart. Given that myocardial ischemic events are often unpredictable in humans, the findings that resveratrol protection is also evident when administered during and/or after the insult adds new dimensions to the clinical potential of resveratrol. PMID- 17115890 TI - Contribution of thioredoxin reductase to T-cell mitogenesis and NF-kappaB DNA binding promoted by selenite. AB - Although the essential role of selenium for cellular immune responses is obvious, delineation of the functions is lacking because selenium can either promote or inhibit cell growth, cytokine production, and activation of transcription factor nuclear factor-kappaB (NF-kappaB). Studies with human thioredoxin-1 (Trx-1) transgenic (Tg) mice were conducted to evaluate the relationship between stimulation of T-cell mitogenic response by sodium selenite and the intracellular Trx-1 levels, and the activities of selenoenzymes and NF-kappaB-DNA binding. Concanavalin A-induced mitogenesis of wild-type mouse splenic cells was stimulated by exposure to low levels of selenite (0.02-0.1 microM), with augmentation of NF-kappaB-DNA binding activity. Treatment with NF-kappaB nuclear translocation inhibitor SN50 or thioredoxin reductase (TR) inhibitor aurothioglucose depressed this stimulatory action. The mitogenic response of Trx 1-Tg mouse splenic cells was enhanced by exposure to relatively high levels of selenite (> or = 0.05 microM), compared with the wild-type mouse. Selenite also augmented TR activity but not cellular glutathione peroxidase activity in the Trx 1-overexpressed cells. These results suggest that the stimulation of T-cell mitogenic response by the physiological levels of selenite is predominantly caused by increased TR activity, which may lead to reduction of Trx-1 dependent on the intracellular expression level and promotion of DNA binding of NF-kappaB. PMID- 17115891 TI - Effect of exercise training on in vitro LDL oxidation and free radical-induced hemolysis: the HERITAGE Family Study. AB - Oxidant stress and overproduction of reactive oxygen species (ROS) contribute to the development of cardiovascular disease. Oxidative modifications of low-density lipoproteins (LDL) are thought to play an early and critical role in atherogenesis. LDL oxidation can be reproduced in vitro, but results usually show a large interindividual variation not entirely explained by the environment. Free radical-induced hemolysis is also proposed to reveal the overall antioxidant capacity. The roles of genetic factors and exercise on the variability of both measures were investigated. The study was conducted in 146 healthy individuals from 28 families participating in a 20-week exercise-training program. In addition to important biological and environmental influences on variation, significant familial aggregation was detected in all oxidation measures. Exercise did not significantly modify the LDL oxidation parameters, but significantly increased resistance was observed in the free radical-induced hemolysis, especially in women, this effect was not observed in smokers. In total, the findings suggest the presence of familial effects in the response to ex vivo oxidation. Further, smoking negates the beneficial effect of exercise training on erythrocyte resistance to free radical-induced hemolysis. These observations emphasize the importance of context in the evaluation of exercise and oxidant stress. PMID- 17115892 TI - Dietary thioproline decreases spontaneous food intake and increases survival and neurological function in mice. AB - Male mice on a diet supplemented with thioproline (l-thiazolidine-4-carboxylic acid), a physiological metabolite of 5-hydroxytryptamine, at 2.0 g/kg of food from 28 weeks of age and for their entire life, showed a 23-29% increased median and maximal life span. These survival increases were associated with improved neurological functions. Compared to control mice, thioproline-supplemented mice had a 20% lower integral spontaneous food intake, and 10% lower body weight at 100 weeks of age. Body weight showed a statistically significant inverse relationship with survival and neurological performances. Thioproline supplemented mice exhibited a 58-70% decrease of the age-dependent oxidative damage in brain and liver mitochondria at 52 weeks (old mice) and 78 weeks (senescent mice) of age, respectively. The age-associated decrease of brain mitochondrial enzyme activities, NADH-dehydrogenase, cytochrome c oxidase, and mitochondrial nitric oxide synthase (mtNOS), in old and senescent mice were markedly prevented (51-74%) by thioproline. In vitro, thioproline neither exhibited direct antioxidant activity nor had any effect on the electron transfer or mtNOS functional activities of brain and liver mitochondria. It is surmised that thioproline induces an anorexic effect associated with improved survival and neurological function through a decreased oxidative damage and regulation that may involve hypothalamic appetite centers. PMID- 17115893 TI - Higher leukocyte 8-oxo-7,8-dihydro-2'-deoxyguanosine and lower plasma ascorbate in aging humans? AB - Is oxidative damage of DNA responsible for physiological changes associated with aging? The authors note a positive correlation between the age of human subjects with the level of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) in leukocyte DNA. The levels of urinary 8-oxo-7,8-dihydroguanine and 8-oxodG followed the same pattern of correlation. Age-dependent decline in the concentration of plasma vitamin C was also evident. These interesting observations in humans point towards the need to scrutinize in detail the role of oxidative DNA damage and compromised antioxidant defense systems in age-related physiological disorders. PMID- 17115894 TI - How does iron-sulfur cluster coordination regulate the activity of human glutaredoxin 2? AB - Human mitochondrial glutaredoxin (Grx2) was described as the first iron-sulfur protein from the thioredoxin superfamily of proteins. The [2Fe-2S] cluster was proposed to serve as redox sensor for the activation of Grx2 during oxidative stress. The authors have demonstrated that the iron-sulfur cluster is complexed by the two N-terminal active site thiols of two Grx2 monomers and two molecules of glutathione that are bound noncovalently to the proteins and in equilibrium with glutathione in solution. When reduced glutathione becomes the limiting factor for cluster coordination, the holo-Grx2 complex dissociates, yielding enzymatically active Grx2. PMID- 17115895 TI - ICAM-1 cross-linking stimulates endothelial glutathione synthesis. AB - What mechanisms regulate endothelial glutathione (GSH) during inflammation? Addressing this question is critical in understanding mechanisms leading to endothelial dysfunction and cardiovascular disease. Herein, the authors show data that support the hypothesis that the intercellular cell adhesion molecule-1 (ICAM 1) regulates GSH. Ligating either constitutive or induced ICAM-1 on the endothelial surface, or exposing endothelial cells to soluble ICAM-1, increases GSH concentrations. ICAM-1 is important in mediating leukocyte adhesion and modulates endothelial signaling pathways important in controlling transmigration. The present data underscore a novel function for ICAM-1 in modulating GSH metabolism and raise the hypothesis that this adhesion molecule controls endothelial redox status under basal and inflammatory conditions. PMID- 17115896 TI - Cytokine production of CD8+ immune T cells but not of CD4+ T cells from Toxoplasma gondii-infected mice is polarized to a type 1 response following stimulation with tachyzoite-infected macrophages. AB - To examine whether cytokine production of CD4(+)immune T cells and CD8(+)immune T cells in Toxoplasma gondii-infected mice differ in their responses to infected cells and to soluble antigens of the parasite, we compared the production of interferon-gamma (IFN-gamma), interleukin-2 (IL-2), IL-4, and IL-10 by the immune T cell populations following in vitro stimulation with tachyzoite-infected macrophages and tachyzoite lysate antigens (TLA). Both CD4(+)and CD8(+)immune T cells produced large amounts of IFN-gamma in response to either infected macrophages or TLA, but the CD4(+)T cells produced greater amounts of the cytokine than did the CD8(+)T cells with both stimulations. Both T cell populations also produced IL-2 after stimulation with infected macrophages, whereas only CD4(+)T cells did when stimulated with TLA. CD4(+)immune T cells also produced large amounts of IL-4 and IL-10 after stimulation with infected macrophages, but CD8(+)T cells did not. These results indicate that CD4(+)immune T cells produce IFN-gamma, IL-2, IL-4, and IL-10 in response to infected macrophages, whereas CD8(+)immune T cells produce predominantly IFN-gamma and IL 2. Since IL-4 and IL-10 could suppress IFN-gamma-mediated protective mechanisms against the parasite, the production of these cytokines by CD4(+)immune T cells in response to infected cells could negatively affect their protective activity in vivo. PMID- 17115897 TI - Thrombin inhibits IFN-gamma production in human peripheral blood mononuclear cells by promoting a Th2 profile. AB - Thrombin, the key enzyme of the coagulation cascade, is involved in inflammation. It was proposed recently that thrombin activity may play an important role in allergic inflammation. Interferon-gamma (IFN-gamma) is a potent Th1-related cytokine secreted by activated T cells and is usually downregulated in allergic inflammation. We recently demonstrated that thrombin enhances interleukin-10 (IL 10) in peripheral blood mononuclear cells (PBMC). Thus, we hypothesized that thrombin may promote a Th2 profile. We here report that human alpha- thrombin downregulates IFN-gamma expression at both protein and mRNA levels in activated PBMCs. The use of proteolytically inactive thrombin and of the specific thrombin receptor agonist peptide, SFLLRN, shows that this downregulation is thrombin specific and requires thrombin proteolytic activity. The addition of an anti- IL 10 monoclonal antibody (mAb) to thrombin-treated PBMCs abolishes IFN-gamma downregulation, suggesting that thrombin exerts its effect through IL-10, a Th2 related cytokine. Furthermore, IFN-gamma reduction was accompanied by increased IL-4 release, as well as by an increase in the proinflammatory cytokine IL-1. In conclusion, the observation that thrombin affects the production of IFN-gamma (Th1 profile) and IL-4 (Th2 profile) provides further evidence for the role played by thrombin in modulating Th1/Th2 cytokine balance, which could be particularly relevant in allergic inflammation. PMID- 17115898 TI - Short communication: phase I clinical and gene modulatory evaluation of tamoxifen and IFN-alpha2b. AB - Preclinical studies had determined that tamoxifen and interferon-alpha2b (IFN alpha2b) synergistically inhibited growth of both estrogen-receptor positive and negative murine tumor xenografts and had combined antiangiogenic effects and that tamoxifen potentiated IFN-stimulated gene (ISG) expression. A phase I trial in 26 patients was conducted using the combination to define tolerance and potentiation of ISG expression. IFN- alpha2b at a dose of 3 x 10(6) units/m(2) daily was given subcutaneously (s.c.), and tamoxifen was initiated as a loading dose of 150 mg/m(2) and then 60 mg/m(2) twice daily on day 8. At this initial dose, reduction of dose of IFN- alpha2b was required in 4 of 11 patients, primarily because of fatigue. Another group of patients was treated with an identical tamoxifen dose but with IFN-alpha2b reduced to 2 x 10(6)/m(2) U; this was better tolerated. As the projected serum tamoxifen level to reduplicate preclinical effects was 300 mg/m(2), dose escalation in a third cohort was undertaken; it had to be discontinued secondary to grade III or IV toxicity in 2 of 2 patients. Increases in products of transcriptionally regulated ISGs, beta (2)-microglobulin, neopterin, and ISG15 were assessed. All ISGs increased after IFN-alpha2b, but only ISG15 had a further significant rise after initiation of tamoxifen. Because at doses not limited by unacceptable toxicities, no marked potentiation of ISGs by tamoxifen could be identified, clinical evaluation of the combination was terminated. PMID- 17115899 TI - Blocking monoclonal antibodies specific for mouse IFN-alpha/beta receptor subunit 1 (IFNAR-1) from mice immunized by in vivo hydrodynamic transfection. AB - Herein we report the generation of mouse monoclonal antibodies (mAbs) specific for the IFNAR-1 subunit of the mouse interferon-alpha/beta (IFN-alpha/beta) receptor (MAR1 mAbs) that block type I IFN receptor signaling and biologic response induction in vitro and in vivo. These mAbs were generated from Ifnar1 (/) mice immunized by in vivo hydrodynamic transfection with a plasmid encoding the extracellular domain (ECD) of murine IFNAR-1. All MAR1 mAbs bound native receptor expressed on cell surfaces and immunoprecipitated IFNAR-1 from solubilized cells, and two mAbs also detected IFNAR-1 by Western blot analysis. in vitro, the mAbs prevented ligand-induced intracellular signaling and induction of a variety of type I IFN-induced biologic responses but had no effect on IFN gamma-induced responses. The most effective in vitro blocker, MAR1-5A3, also blocked type I IFN-induced antiviral, antimicrobial, and antitumor responses in vivo. We also explored whether murine IFNAR-1 surface expression required the presence of Tyk2. In contrast to Tyk2-deficient human cell lines, comparable IFNAR-1 expression was found on primary cells derived either from wild-type or Tyk2 (/) mice. These mAbs represent much needed tools to more clearly elucidate the biochemistry, cell biology, and physiologic function of the type I IFNs and their receptor in mediating host-protective immunity and immunopathology. PMID- 17115900 TI - An alternative spliced RNASEL variant in peripheral blood leukocytes. AB - 2-5A-Dependent RNase L is an endoribonuclease that catalyzes RNA degradation and promotes apoptosis during the innate antiviral response in mammalian cells. Prior studies showed that RNASEL is widely expressed and suggested the presence of mRNA species of different sizes but lacked a characterization of these variants. Using RT-PCR, we show that RNASEL is expressed in all human tissues examined, whereas an alternatively generated spliced variant lacking the third exon (RNASEL del_Ex3) is solely expressed in peripheral blood leukocytes (PBL). Quantitative RT-PCR measurements of RNA from different PBL cell types separated by fluorescence activated cell sorting (FACS) showed that complete RNASEL mRNA levels were significantly elevated in granulocytes compared with all other PBL cell types, whereas expression was lowest in CD8(+) T cells. The alternatively spliced RNASEL del_Ex3 transcript was present in all PBL cell types examined but at lower levels than the full-length RNASEL mRNA. The presence of high levels of RNase L protein in granulocytes was confirmed by immunohistochemistry. Our findings are the first to demonstrate the presence of an alternatively spliced RNASEL mRNA and to demonstrate the variable expression of RNase L in different leukocytes. Our results suggest that RNase L plays an important role in granulocytes as an innate immunity enzyme that controls viral infections. PMID- 17115901 TI - Short communication: differences between macrophages and dendritic cells in the cyclic AMP-dependent regulation of lipopolysaccharide-induced cytokine and chemokine synthesis. AB - Cyclic adenosine monophosphate (cAMP) is an intracellular signaling molecule responsible for directing cellular responses to extracellular signals. Once believed to signal exclusively through its ability to bind protein kinase A (PKA), recent research has revealed alternative cAMP-binding targets involved in PKA-independent processes. In this study we addressed the hypothesis that the guanine nucleotide exchange protein directly activated by cAMP (Epac-1) and PKA differentially regulate inflammatory mediator production in distinct phagocytic cell types. To accomplish this, we compared the release of cAMP-regulated polypeptide inflammatory mediators in both macrophages (obtained from the lung and peritoneum) and bone marrow-derived dendritic cells (DCs) stimulated with bacterial endotoxin. Using the highly selective Epac-1 and PKA activating cAMP analogs 8-pCPT-2 -O-Me-cAMP and 6-Bnz-cAMP, respectively, we found that macrophages differ from DCs in the involvement of these distinct cAMP pathways in modulating inflammatory mediator release in response to endotoxin. Whereas the regulation of cytokine and chemokine production in macrophages by cAMP was solely dependent on PKA, we found that both Epac-1 and PKA activation could regulate mediator production in DCs. This finding may be important in the pharmacologic regulation of immune responses through manipulation of cAMP signaling cascades and contributes to our understanding of the differences between these cell types. PMID- 17115902 TI - Effects of HCV treatment on cytokine expression during HCV/HIV coinfection. AB - There is growing evidence that cytokine expression is linked to hepatitis C virus (HCV) pathogenesis and treatment response rates among HCV-monoinfected persons. However, because of the profound effects of human immunodeficiency virus (HIV) coinfection on HCV, it is not clear if these observations are also true for HCV/HIV-coinfected individuals. Serum expression of the proinflammatory cytokines interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-alpha) and the fibrogenic cytokine transforming growth factor-beta1 (TGF-beta1) were measured in HCV/HIV-coinfected persons at baseline and at week 24 of HCV therapy. Higher levels of IL-8 and TGF-beta were demonstrated among nonwhite subjects at baseline. Increases in TNF-alpha and IL-8 expression were found at week 24 of HCV therapy, suggesting that enhanced proinflammatory cytokine production may occur during HCV treatment. However, cytokine levels were not predictive of HCV virologic, biochemical, or histologic response. Although previous studies conducted among HCV-monoinfected individuals have suggested that cytokine levels could predict the virologic response to therapy, no such associations were observed among HCV/HIV-coinfected persons, suggesting that they may respond differently to treatment than do their HCV-monoinfected counterparts. PMID- 17115904 TI - A note from the editors: manuscript retraction. PMID- 17115905 TI - Proteolysis consistent with activation of caspase-7 after severe traumatic brain injury in humans. AB - The expression and proteolysis of caspase family proteins are involved in the initiation and execution of apoptosis, which has been reported to occur in human and experimental traumatic brain injury (TBI). Caspase-3, -6, and -7 belong to the group of executioner caspases, which are cleaved and activated at the late, irreversible stage of apoptosis. Our previous studies demonstrated roles for caspase-1, -3, and -8 in humans after severe TBI. Here we report expression of caspase-7 mRNA and protein in humans after TBI (n = 16) and control brain-bank tissue (n = 6). Semiquantitative reverse transcription polymerase chain reaction showed no differences between caspase-7 mRNA in TBI patients versus controls (73 +/- 24 vs. 85 +/- 56 relative optical density [ROD], respectively). In contrast, Western blot analysis showed increased pro-caspase-7 in TBI patients versus controls (214 +/- 30 vs. 1 +/- 1 ROD, respectively), as well as an increase in the approximately 20 kD proteolytic fragment in TBI patients versus controls (86 +/- 13 vs. 22 +/- 12 ROD, respectively), consistent with activation of caspase-7 after TBI in humans. Immunohistochemical analysis showed that cells expressing caspase-7 included astrocytes and neurons and possibly other glial cell types and infiltrated inflammatory cells. These data show that caspase-7 and its cleavage product are increased in human brain after TBI in many central nervous system, as well as noncentral nervous system, cell types. Thus, caspase-7 may play a role in the glial and inflammatory responses, and possibly neuronal death, after TBI in humans. PMID- 17115906 TI - Alkoxyl radical-scavenging activity of edaravone in patients with traumatic brain injury. AB - Lipid peroxidation is caused by reactive oxygen species (ROS) and is involved in traumatic brain injury (TBI). Consequently, a therapeutic strategy for TBI may be to control lipid peroxidation. The only drug approved to date for blocking lipid peroxidation is edaravone (MCI-186), a novel free-radical scavenger shown to exert neuroprotective effects in acute ischemic stroke. Although edaravone scavenges hydroxyl and nitric oxide radicals, its effect on alkoxyl radicals (OR ), which also contribute to lipid peroxidation, is unknown. To date, the study of free radicals in blood has been severely hampered by technical difficulties in their detection. We used an in vitro and ex vivo electron spin resonance (ESR) method employing 5,5-dimethyl-1-pyrroline-N-oxide as a spin trap to investigate whether edaravone can scavenge OR-. By mixing either methemoglobin or human blood with tert-butyl hydroperoxide, we found that this technique can detect OR- generated in vitro. We also found that generated OR- can be completely absorbed by administration of edaravone in vitro (400 microM). Analysis of jugular venous blood collected from 17 TBI patients immediately before and 20 minutes after the administration of edaravone (30 mg, i.v.) revealed higher OR- levels in the untreated patients blood than in normal control blood samples. However, treatment with edaravone suppressed these OR- levels by 24.6% (radical intensity = 71.1 +/- 5.2-53.6 +/- 5.2; p < 0.01). Thus, edaravone can scavenge OR- and significantly reduce levels of these radicals in TBI patients. The novel ex vivo ESR method described here provides a valuable clinical measure of oxidative stress. PMID- 17115907 TI - Outcome of traumatic brain injury after three decades--relationship to ApoE genotype. AB - Significant traumatic brain injury (TBI) is nearly always associated with cognitive deficits, but in a highly variable manner. Apolipoprotein E (ApoE) plays a pivotal role in CNS response to injury. To examine the association of ApoE genotype with long-term outcome in TBI patients, we determined the ApoE genotype from 61 TBI patients who had been injured over three decades earlier. All patients had been studied neuropsychologically after their injuries. The long term outcome was evaluated with repeated neuropsychological testing and by applying various measures of everyday functioning and quality of life. After three decades, TBI patients with the ApoE epsilon4 allele showed significantly poorer general cognitive level than those without this allele. This decline was wholly accounted for by a subgroup of these patients who had developed incident or clinical dementia, while the majority of the ApoE epsilon4 positive patients showed no decline at all. The other outcome measures describing vocational, physical, or subjective symptom outcome did not show significant relationships to the ApoE genotype. A portion of the TBI patients with the ApoE epsilon4 allele seem to be at risk of long-term cognitive decline. PMID- 17115908 TI - Use of the WHOQOL-BREF for evaluating persons with traumatic brain injury. AB - This study examined psychometric properties of a brief version of the World Health Organization Quality of Life questionnaire (WHOQOL-BREF) among persons with traumatic brain injury (TBI) and the relations of the WHOQOL-BREF domains, including physical capacity, psychological wellbeing, social relationships, and environment, to different indicators of TBI severity. Of the 354 eligible and available subjects from 22 hospitals in northern Taiwan over a 6-month period, 199 completed telephone interviews during data collection. Three indicators of TBI severity were used: the Glasgow Coma Scale, the presence of post-traumatic amnesia, and the abbreviated injury scale to the head. All domain scores of the WHOQOL-BREF had nearly symmetrical distributions: low percentages of ceiling and floor values (0-3%), low missing rates (0-0.5%) for all but one item (43.2%), and very good internal consistency (0.75-0.89) and test-retest reliability (0.74 0.95). The WHOQOL-BREF also exhibited excellent known-groups validity, as well as very good responsiveness and convergent validity with regard to employment, independence in daily life activities, social support, and depression. After adjustment for potential confounders, almost none of the domain scores of the WHOQOL-BREF significantly differed in the severity levels of the three severity indicators. In conclusion, the WHOQOL-BREF is an appropriate health-related quality of life (HRQL) instrument for persons with TBI. Furthermore, the initial severity of the TBI might not be suitable for predicting levels of HRQL in persons with TBI. PMID- 17115909 TI - Fighting for each segment: estimating the clinical value of cervical and thoracic segments in SCI. AB - Patients suffering from complete spinal cord injury (SCI) are the most likely candidates for the application of new interventions for neural repair and regeneration. It is assumed that some of these treatments will have their strongest impact at the segmental level. Therefore, it is important to evaluate the clinical relevance of potential changes at the segmental levels concerning both improvement and deterioration. Data of 98 motor complete SCI patients were derived from the European Multicenter Study of Human Spinal Cord Injury database. Six months after injury, the ASIA motor score and Spinal Cord Independence Measure (SCIM) were assessed as dependent variables (linear regression analysis) to disclose the difference between each segment. Separate analyses using linear regression for tetraplegic patients (n = 39) and paraplegic patients with thoracic lesions (n = 54) were performed to calculate the difference between each spinal segment. In tetraplegic patients, both the ASIA motor score and the SCIM revealed relevant differences per spinal segment (9 and 4 points, respectively) while in paraplegic patients there was no difference for the SCIM and the ASIA motor score between T2 and T8. We suggest that in complete tetraplegic patients, changes of even one spinal segment will either improve or degrade both motor function and independence. Segmental changes at the thoracic level are not assessable by the ASIA motor score and SCIM tests. Therefore, the assessment of efficacy and safety in thoracic patients by these two tests has limited value when applied to cervical SCI. These findings may be considered in clinical trials for the evaluation of beneficial effects and risk management when treating patients with spinal cord injury. PMID- 17115910 TI - Quantification of locomotor recovery following spinal cord contusion in adult rats. AB - Injury to the spinal cord not only disrupts the functioning of spinal circuits at the site of the impact, but also limits sensorimotor function caudal to the level of the lesion. Ratings of gross locomotor skill are generally used to quantify locomotor recovery following spinal cord injury (SCI). The purpose of this study was to assess behavioral recovery following SCI with three tasks: (1) BBB ratings, (2) walking on a horizontal ladder, and (3) footprint analyses. Behavioral testing was conducted for 6 postoperative weeks, and then the spinal cords were processed for the amount of white matter spared. As expected, BBB ratings dramatically decreased and then improved during recovery. The number of hindlimb foot-faults on the horizontal ladder increased after injury and remained elevated during the recovery period. Footprint analyses revealed that sham control rats used several different gaits to cross the runway. In contrast, the locomotor function of rats with a SCI was impaired throughout the postoperative period. Some locomotor parameters of the injured rats improved slightly (velocity, stride length, stride duration, stance duration), some did not change (interlimb coordination, swing duration, forelimb base of support, hindpaw angle), and others declined (hindlimb base of support) during the recovery period. Together, these results show that gross locomotor skill improved after SCI, while recovery of fine locomotor function was more limited. Multiple tests should be included in future experiments in order to assess gross and fine changes in sensorimotor function following SCI. PMID- 17115911 TI - The Louisville Swim Scale: a novel assessment of hindlimb function following spinal cord injury in adult rats. AB - The majority of animal studies examining the recovery of function following spinal cord injury use the BBB Open-Field Locomotor Scale as a primary outcome measure. However, it is now well known that rehabilitation strategies can bring about significant improvements in hindlimb function in some animal models. Thus, improvements in walking following spinal cord injury in rats may be influenced by differences in activity levels and housing conditions during the first few weeks post-injury. Swimming is a natural form of locomotion that animals are not normally exposed to in the laboratory setting. We hypothesized that deficits in, and functional recovery of, swimming would accurately represent the locomotor capability of the nervous system in the absence of any retraining effects. To test this hypothesis, we have compared the recovery of walking and swimming in rats following a range of standardized spinal cord injuries and two different retraining strategies. In order to assess swimming, we developed a rating system we call the Louisville Swimming Scale (LSS) that evaluates three characteristics of swimming that are highly altered by spinal cord injury--namely, hindlimb movement, forelimb dependency, and body position. The data indicate that the LSS is a sensitive and reliable method of determining swimming ability and the improvement in hindlimb function after standardized contusion injury of the thoracic spinal cord. Furthermore, the data suggests that when used in conjunction with the BBB Open-field Locomotor Scale, the LSS assesses locomotor capabilities that are not influenced by a retraining effect. PMID- 17115912 TI - Changes in soleus muscle function and fiber morphology with one week of locomotor training in spinal cord contusion injured rats. AB - The purpose of this study is two-fold: (1) to examine skeletal muscle function in a rat model of midthoracic contusion spinal cord injury (SCI) and (2) to evaluate the therapeutic influence of a short bout (1 week) of treadmill locomotor training on soleus muscle function (peak force, fatigability, contractile properties, fiber types), size (fiber area), and motor deficit and recovery (BBB scores) after SCI. The rats were injured with a moderate T8 spinal cord contusion and were assigned to either receive treadmill locomotor training (TM), starting 1 week after SCI for 5 consecutive days (20 min/trial, 2 trials/day) or not to receive any exercise intervention (no TM). Locomotor training resulted in a significant improvement in overall locomotor function (32% improvement in BBB scores) when compared to no TM. Also, the injured animals that trained for 1 week had 38% greater peak soleus tetanic forces (p < 0.05), a 9% decrease in muscle fatigue (p < 0.05), 23% larger muscle fiber CSA (p < 0.05), and decreased immunoexpression of fast heavy chain fiber types than did rats receiving no TM. In addition, there was a good correlation (0.704) between the BBB scores of injured animals and peak soleus muscle force regardless of group assignment. No significant differences were seen in twitch or time to peak tension values across groups. Collectively, these results indicate that 1 week of treadmill locomotor training, initiated early after SCI, can significantly improve motor recovery following SCI. The magnitude of these changes is remarkable considering the relatively short training interval and clearly illustrates the potential that initiating treadmill locomotor training shortly after injury may have on countering some of the functional deficits resulting from SCI. PMID- 17115913 TI - Bone marrow stromal cell transplantation preserves gammaaminobutyric acid receptor function in the injured spinal cord. AB - A surprising shortage of information surrounds the mechanisms by which bone marrow stromal cells (BMSC) restore lost neurologic functions when transplanted into the damaged central nervous system. In the present study, we sought to elucidate whether BMSCs express the neuron-specific gamma-aminobutyric acid (GABA) receptor when transplanted into injured spinal cord. To examine this, we harvested and cultured rat femoral BMSCs. We then subjected Sprague-Dawley rats to thoracic spinal cord injury (SCI) with a pneumatic impact device. Fluorescence labeled BMSCs (n = 7) were transplanted stereotactically or the vehicle in which these cells were cultured (n = 4) was introduced stereotactically into the rostral site of SCI at 7 days after injury. We evaluated GABA receptor function by measuring the binding potential for 125I-iomazenil (125I-IMZ) through in vitro autoradiography at 4 weeks after BMSC transplantation and simultaneously examined the fate of the transplanted BMSCs by immunocytochemistry. We found that the transplanted BMSC migrated toward the core of the injury and were densely distributed in the marginal region at 4 weeks after transplantation. BMSC transplantation significantly increased the binding potential for 125I-IMZ (p = 0.0376) and increased the number of GABA receptor-positive cells (p = 0.0077) in the marginal region of the injury site. Some of the transplanted BMSCs were positive for microtubule-associated protein-2 and the alpha1 subunit of GABA(A) receptor in the region of injury. These findings suggest that BMSCs have the potential to support the survival of neurons in the marginal region of SCI and can partly differentiate into neurons, regenerating spinal cord tissue at the site of injury. PMID- 17115914 TI - Effect of oxidative stress on glial cell volume. AB - Cytotoxic brain edema is a major contributor of tissue damage following cerebral ischemia and traumatic brain injury. The pathophysiology of cytotoxic edema formation is still not well understood. Although it is widely believed that oxidative stress causes cytotoxic brain edema, experimental proof is lacking. The aim of the present study was therefore to examine the effect of oxidative stress on cell volume of glial cells. C6 glial cells were exposed to hydrogen peroxide and the superoxide forming complex hypoxanthine/xanthine oxidase (HX/XO). Exposure to hydrogen peroxide (0.5-5 mM) resulted in initial cell shrinkage by 5.7 +/- 1.5% (mean +/- SEM; p < 0.05) and was followed by a dose-dependent recovery to baseline. Exposure to superoxide anions generated by HX/XO provoked a delayed, but sustained decrease of cell volume by 11.8 +/- 0.9% (p < 0.05). Cell volume showed no tendency to recover upon sustained exposure to superoxide. Neither hydrogen peroxide nor HX/XO exposure was associated with a decrease of cell viability. Thereby, the present study demonstrates that oxidative stress by hydrogen peroxide and superoxide anions does not induce cytotoxic cell swelling and suggests that free radicals are not directly involved in the formation of cytotoxic brain edema. PMID- 17115915 TI - Corticospinal tract transection permanently abolishes H-reflex down-conditioning in rats. AB - Previous studies have shown that corticospinal tract (CST) transection, but not transection of other major spinal cord tracts, prevents down-conditioning of the H-reflex, the electrical analog of the spinal stretch reflex. This study set out to determine whether the loss of the capacity for H-reflex down-conditioning caused by CST transection is permanent. Female Sprague-Dawley rats received CST, lateral column (LC), or dorsal column ascending tract (DA) transection at T8-9; 9 10 months later, they were exposed to the H-reflex down-conditioning protocol for 50 days. In the LC and DA rats, H-reflex size fell to 60 (+/- 9 SEM)% and 60 (+/- 19)%, respectively, of its initial size. This down-conditioning was comparable to that of normal rats. In contrast, H-reflex size in the CST rats rose to 170 (+/- 42)% of its initial size. A similar rise does not occur in rats exposed to down conditioning shortly after CST transection. These results indicate that CST transection permanently eliminates the capacity for H-reflex down-conditioning and has gradual long-term effects on sensorimotor cortex function. They imply that H-reflex down-conditioning can be a reliable measure of CST function for long-term studies of the effects of spinal cord injury and/or for evaluations of the efficacy of experimental therapeutic procedures, such as those intended to promote CST regeneration. The results also suggest that the role of sensorimotor cortex in down-conditioning extends beyond generation of the essential CST activity. PMID- 17115921 TI - An interview with Richard M. Eglen, Ph.D. PMID- 17115923 TI - A cellular assay for measuring the modulation of glucose production in H4IIE cells. AB - Type II diabetes and its associated complications are a major health concern of the developed world. One of the hallmarks of diabetes is insulin resistance, where secreted insulin no longer has any effect on its target tissues, namely, liver, muscle, and fat. An important therapeutic strategy is to modulate blood glucose levels using pharmacological agents. Glycogen synthase kinase-3 (GSK3) is a serine-threonine protein kinase that plays important roles in regulating glucose metabolism. It is a key negative regulator of insulin action and is an important contributing factor to insulin resistance in liver, muscle, and adipose tissue. We describe the development of a cell-based assay designed to measure glucose production in rat hepatoma cell line H4IIE liver cells in response to treatment with small molecule inhibitors, including GSK3 inhibitors. The assay is set up in a 96-well format, and glucose production is assessed using a convenient fluorescence-based readout. This disease-relevant cellular assay is a valuable tool for the progression of small molecules that modulate glucose production. PMID- 17115924 TI - Biotinylated peptides for rapid identification of substrates and inhibitors of kinases and phosphatases with fluorescence superquenching. AB - Aberrant regulation of kinase and phosphatase activities is implicated in various diseases, including cancer, diabetes, and inflammation. Thus, high-throughput screening (HTS) has become a focused strategy for the identification of kinase and phosphatase inhibitors. With a growing number of these enzymes becoming available for HTS, rapid identification of substrates has become pertinent. Several substrate panel screening assays exist that allow the researcher to test dye-labeled peptides for kinase or phosphatase activity. Here we introduce a method that uses readily available biotinylated peptides instead of dye-labeled substrates, which are costly and limited in availability. After enzymatic phosphorylation, biotinylated peptides are coupled to streptavidin-quencher conjugates, which then associate with a fluorescent polymer via phosphate-metal ion interaction between the reacted biotinylated peptide complex and the polymer. As a result, quencher and polymer are brought into a proximity that allows electron transfer from the polymer to the dye. The Dylight(647) (Pierce, Rockford, IL) dye was identified as an efficient electron transfer molecule that allows assays to be monitored using two emission wavelengths simultaneously, 490 nm from the polymer and 685 nm from the transferred emission of the dye. Assays are homogeneous and show comparable sensitivities to assays performed with direct labeled dyes. When applied to a limited screen using previously characterized peptides, substrates for two kinases and one phosphatase were correctly identified. Further, ratiometric analysis of polymer quenching and transferred emission accurately detected inhibitors in a compound screen against protein kinase A, protein kinase Calpha, and protein tyrosine phosphatase 1B with limited interferences from colored compounds and with Z factors of >0.7. PMID- 17115925 TI - Live cell quality control and utility of real-time cell electronic sensing for assay development. AB - In this paper we have explored the utility of the real-time cell electronic sensing (RTCES, ACEA Biosciences Inc., San Diego, CA) system for monitoring the quality of live cells in cell-based assays as well as for assay development. We have demonstrated that each cell type displays unique growth kinetic profiles that provide a quantitative account of cell behavior and can be used as a diagnostic tool for cellular quality control. The utility of the specific signature patterns was shown by demonstrating the significant differences in primary cell behavior depending on the supplier. In addition, the RT-CES system was able to differentiate cell behavior depending on the passage stage of the cells. The utility of the RT-CES system as an assay development tool was demonstrated in cytotoxicity assays. The RT-CES system not only provides information regarding the potency of cytotoxic compounds, but in addition relates potency to the rate of the response for each concentration of the compound tested, which is important for understanding the mechanism of compound action. Moreover, real-time display of cytotoxicity data by the RT-CES system allows for calculation of real-time 50% inhibitory concentration (IC50) values or determination of optimal IC(50) value. In summary, the RT-CES system provides high content and information-rich data that are beyond the scope of single-point assays. PMID- 17115926 TI - Dynamic detection of natural killer cell-mediated cytotoxicity and cell adhesion by electrical impedance measurements. AB - Measurement of electrical impedance is a relatively new real-time and label-free method for monitoring cell adhesive properties. Impedance measurements are performed in tissue culture wells in which the bottom is equipped with gold electrodes. The extent of electrode coverage by living cells as well as the strength of the bond between the cell membrane and the electrode surface determines the impedance, which in real-time cell electrical sensing (RT-CES, ACEA Biosciences, San Diego, CA) is measured as the cell index (CI). We showed for carcinoma cells that CI was linearly correlated to the number of cells and that CI also was related to the amount of coating (laminin-5) of the wells. When natural killer (NK) cells were added to adherent carcinoma cells (target cells) CI declined rapidly dependent on the NK cell:target cell ratio. The initial decrease of CI was much more pronounced than target cell death as measured by [(3)H]thymidine incorporation assay. Such a rapid fall of CI was due to changes in the adhesion and morphology of target cell undergoing apoptosis. It took more than 6 h before the extent of cell death and fall of CI were comparable. We also showed using A431 cells and an antibody specific for the human epidermal growth factor receptor (Erbitux, manufactured by Merck KGaA, Darmstadt, Germany) that RT CES could be used to monitor antibody-dependent cellular cytotoxicity. Thus RT CES is a convenient way to continuously determine cell number and cell adhesion and may offer early detection of NK cell-mediated cytotoxic effects. PMID- 17115927 TI - Quantification of small molecule-receptor affinities and kinetics by acoustic profiling. AB - The label-free RAPid 4 system that exploits resonant acoustic profiling (RAP) from Akubio (Cambridge, UK) was used to determine the affinity and kinetics for several different small molecule-receptor interactions. This was achieved by attaching the target receptor to the surface of quartz crystal resonators through a variety of specific coupling chemistries, followed by application of a small molecular-weight ligand to the receptor via a microfluidic flow-based delivery system. Rank order of binding was determined for very weak interactions such as cofactor binding to glucose dehydrogenase. Moderate interaction affinities and binding kinetics could be determined for biotin binding to a specific antibody, and also for several low-molecular-weight sulfonamide analogues binding to human carbonic anhydrase isoform II. The equilibrium binding constants were in general agreement with the values obtained by kinetic analysis of the data, as well as with previously published values obtained using surface plasmon resonance, stopped flow fluorescence, and isothermal titration calorimetry. PMID- 17115928 TI - Transporter assays using solid supported membranes: a novel screening platform for drug discovery. AB - Transporters are important targets in drug discovery. However, high throughput capable assays for this class of membrane proteins are still missing. Here we present a novel drug discovery platform technology based on solid supported membranes. The functional principles of the technology are described, and a sample selection of transporter assays is discussed: the H(+)-dependent peptide transporter PepT1, the gastric proton pump, and the Na(+)/Ca(2+) exchanger. This technology promises to have an important impact on the drug discovery process. PMID- 17115929 TI - Label-free cell-based assays with optical biosensors in drug discovery. AB - Once viewed solely as a tool for low throughput and kinetic analysis of biomolecular interactions, optical biosensors are gaining widespread uses in drug discovery because of recent advances in instrumentation and experimental design. These advances have expanded the capabilities of optical biosensors to meet the needs at many points in the drug discovery process. Concurrent shifts in drug discovery paradigms have seen the growing use of whole cell systems for drug screens, thus creating both a need in drug discovery and a solution in optical biosensors. This article reviews important advances in optical biosensor instrumentation, and highlights the potential of optical biosensors for drug discovery with an emphasis on whole cell sensing in both high throughput and high content fashions. PMID- 17115930 TI - Dynamic and label-free cell-based assays using the real-time cell electronic sensing system. AB - Cell-based assays have become an integral part of the preclinical drug development process. Recently, noninvasive label-free cell-based assay technologies have taken center stage, offering important and distinct advantages over and in addition to traditional label-based endpoint assays. Dynamic monitoring of live cells, the preclusion of label, and kinetics are some of the fundamental features of cell-based label-free technologies. In this article we will discuss the real-time cell electronic sensing (RT-CES, ACEA Biosciences Inc., San Diego, CA) system and some of its key applications for cell-based assays such as cell proliferation and cytotoxicity, functional assays for receptor-ligand analysis, cell adhesion and spreading assays, dynamic monitoring of endothelial barrier function, and dynamic monitoring of cell migration and invasion. Also, where appropriate we will briefly discuss other label-free technologies in an application-specific manner. PMID- 17115931 TI - Cellular dielectric spectroscopy: a label-free comprehensive platform for functional evaluation of endogenous receptors. AB - The CellKey (MDS Sciex, South San Francisco, CA) system enables comprehensive pharmacological evaluation of cell surface receptors, including G-protein coupled receptors (GPCRs) and tyrosine kinase receptors, using adherent and suspension cell lines and primary cells. A unique application enabled by the ability of the CellKey system to reliably quantify activation of endogenous receptors is receptor panning. This application allows investigators to easily screen disease relevant cell types for functionally active target receptors by treating cells with a panel of receptor-specific ligands. Receptor panning of multiple cell types including Chinese hamster ovary, human embryonic kidney 293, HeLa, U-937, U 2 OS, and TE671 cells resulted in the identification of many functionally active, differently coupled endogenous GPCRs, some of which have not been previously documented in the literature. Upon detecting GPCR activation in live cells, unique cellular dielectric spectroscopy (CDS) response profiles are generated within minutes that reflect the signaling pathways utilized and have been shown to be characteristic of Gs, Gq, and Gi GPCRs. The fact that the CDS response profiles are predictive of the G-protein coupling mechanism of the receptor was demonstrated by using examples of subtype-selective agonists/antagonists to identify the subtypes of the endogenous histamine and beta-adrenergic receptors expressed in U-2 OS cells. A direct correlation is shown between receptor subtype G-protein coupling and CDS response profile. In addition, complex pharmacology, including detection of partial agonism and Schild analysis for endogenous receptors, is presented. The CellKey system allows investigators to conduct studies using endogenously expressed receptors to generate data that are physiologically relevant and in disease context. PMID- 17115933 TI - Glucose-transport regulation in leukemic cells: how can H2O2 mimic stem cell factor effects? AB - In leukemic cells, glucose transport is activated by SCF and H2O2 through a common signal cascade involving Akt, PLCgamma, Syk, and the Src family, in this order. An explanation can be provided by the phosphorylation of c-kit, the SCF receptor, elicited by either SCF or H2O2. Moreover, antioxidants prevent the SCF effect on glucose transport, confirming the involvement of H2O2 in the pathway leading to glucose-transport activation and suggesting a potential role for reactive oxygen species in leukemia proliferation. PMID- 17115934 TI - Superoxide dismutase and hippocampal function: age and isozyme matter. AB - Superoxide dismutases (SODs) are the major antioxidant enzymes that inactivate superoxide and thereby control oxidative stress as well as redox signaling. Transgenic mice overexpressing different isozymes of SOD have been used to study the effect of SOD overexpression on hippocampal synaptic plasticity and hippocampus-dependent learning and memory. Studies with transgenic and wild-type animals of different ages show that the function of SOD overexpression changes across the life span of an animal, and comparisons between animals that overexpress different SOD isozymes suggest that the functional value of overexpression as well as the mechanisms through which the respective functional values are effected vary depending on isozyme. The work discussed in this review has important implications for the use of antioxidant treatments and for our understanding of the role of superoxide in physiological and pathological processes. PMID- 17115935 TI - Ca2+ storage capacity of rat brain mitochondria declines during the postnatal development without change in ROS production capacity. AB - Ca2+ overload of mitochondria and oxidants are considered as crucial factors inducing the opening of the permeability transition pore (PTP) in mitochondria. The interdependence between permeability transition (PT), calcium retention capacity (CRC), and reactive oxygen species (ROS) generation was studied in mitochondria from immature and mature rat brain. Brain mitochondria isolated from 1-day- and 1-week-old rats are much more resistant to Ca2+-triggered PT in phosphate-containing incubation medium than mitochondria from adult brain, since the CRC decreases with development. CRC of mitochondria from 1-week-old rat brain was higher than for adult rat brain (450 +/- 112 vs. 175 +/- 35 nmol Ca2+ per mg of protein). In contrast, for ROS generation there was no age difference. In immature and mature mitochondria, basal, respiratory chain-inhibited or glutathione-depleted ROS generations were similar. In addition, the extent of the Ca2+ load was without effect on the basal ROS generation before mitochondria underwent PT. In summary, ROS generation does not crucially affect the ability of immature mitochondria to buffer high levels of extramitochondrial Ca2+ without undergoing PT. However, we hypothesize that the high resistance of immature mitochondria is related to the low content of some PTP complex constituents, such as creatine kinase. PMID- 17115937 TI - A role for reactive oxygen/nitrogen species and iron on neuronal synaptic plasticity. AB - A great body of experimental evidence collected over many years indicates that calcium has a central role in a variety of neuronal functions. In particular, calcium participates in synaptic plasticity, a neuronal process presumably correlated with cognitive brain functions such as learning and memory. In contrast, only recently, evidence has begun to emerge supporting a physiological role of reactive oxygen (ROS) and nitrogen (RNS) species in synaptic plasticity. This subject will be the central topic of this review. The authors also present recent results showing that, in hippocampal neurons, ROS/RNS, including ROS generated by iron through the Fenton reaction, stimulate ryanodine receptor mediated calcium release, and how the resulting calcium signals activate the signaling cascades that lead to the transcription of genes known to participate in synaptic plasticity. They discuss the possible participation of ryanodine receptors jointly stimulated by calcium and ROS/RNS in the normal signaling cascades needed for synaptic plasticity, and how too much ROS production may contribute to neurodegeneration via excessive calcium release. In addition, the dual role of iron as a necessary, but potentially toxic, element for normal neuronal function is discussed. PMID- 17115936 TI - Sources and targets of reactive oxygen species in synaptic plasticity and memory. AB - Increasing evidence suggests that reactive oxygen species (ROS), such as superoxide and hydrogen peroxide, act as necessary signaling molecules in processes underlying cognition. Moreover, ROS have been shown to be necessary in molecular process underlying signal transduction, synaptic plasticity, and memory formation. Research from several laboratories suggests that NADPH oxidase is an important source of superoxide in the brain. Evidence is presented here to show that ROS are in fact important signaling molecules involved in synaptic plasticity and memory formation. Moreover, evidence that the NADPH oxidase complex is a key regulator of ROS generation in synaptic plasticity and memory formation is discussed. Understanding redox signaling in the brain, including the sources and molecular targets of ROS, are important for a full understanding of the signaling pathways that underlie synaptic plasticity and memory. Knowledge of ROS function in the brain also is critical for understanding aging and neurodegenerative diseases of the brain given that several of these disorders, including Alzheimer's disease and Parkinson disease, may be exacerbated by the unregulated generation of ROS. PMID- 17115938 TI - Differentiated astrocytes acquire sensitivity to hydrogen sulfide that is diminished by the transformation into reactive astrocytes. AB - Hydrogen sulfide (H2S) enhances the induction of hippocampal long-term potentiation (LTP) and induces calcium waves in astrocytes. Based on these observations, H2S has been proposed to be a synaptic modulator in the brain. Here we show that differentiated astrocytes acquire sensitivity to H2S that is diminished by their transformation into reactive astrocytes. Although sodium hydrosulfide hydrate (NaHS), a donor of H2S, did not increase the intracellular concentration of Ca2+ in progenitors, exposure of progenitors to leukemia inhibitory factor (LIF), which induces differentiation into glial fibrillary acidic protein (GFAP)-positive astrocytes, greatly increased the sensitivity to NaHS. In contrast, epidermal growth factor (EGF), transforming growth factor alpha (TGF-alpha), dibutyryl cyclic AMP (db cAMP) and interleukin-1beta (IL 1beta) induced the conversion to reactive astrocytes with diminished sensitivity to NaHS. This suppressive effect of EGF on the sensitivity to NaHS was inhibited by cycloheximide, indicating that de novo protein synthesis was required for the suppression of H2S sensitivity. PMID- 17115939 TI - Aged SOD overexpressing mice exhibit enhanced spatial memory while lacking hippocampal neurogenesis. AB - The recent finding that hippocampal slices from aged mice overexpressing the gene for superoxide dismutase (SOD) exhibit long-term potentiation (LTP) of reactivity to afferent stimulation that is significantly larger than that produced in aged wild-type (wt) mice has encouraged the exploration of the effects of reactive oxygen species (ROS) on learning in aged mice. In addition, young-adult and aged wt and SOD transgenic mice were used in an attempt to correlate adult neurogenesis with spatial learning. Aged wt and SOD mice exhibited a 90% reduction in doublecortin-labeled new dentate gyrus neurons as compared to young mice, with no significant difference between genotypes. In addition, aged SOD mice exhibited better performance than wt controls in both reference and working memory tasks in a water maze. These findings provide a behavioral measure to demonstrate that excessive production of H2O2 is beneficial in aged mice. PMID- 17115940 TI - The contribution of the DNA damage response to neuronal viability. AB - Neurons are extremely active cells and metabolize up to 20% of the oxygen that was consumed by the organism. Despite their highly oxygenic metabolism, neuronal cells have a lower capacity to neutralize the reactive oxygen species (ROS) that they generate or to which they are exposed. High levels of ROS can lead to accumulation of damage to various cellular macromolecules. One of the cellular macromolecules highly affected by intracellular as well as extracellular insults is DNA. Neurons are also highly differentiated, postmitotic cells that cannot be replenished after disease or trauma. Since neurons are irreplaceable and should survive as long as the organism does, they need elaborate defense mechanisms to ensure their longevity. This review article mainly focuses on certain mechanisms that contribute to neuronal longevity, and concentrates on the DNA damage response in neuronal cells. The various mechanisms of DNA repair are briefly described, and focus is on those mechanisms that are activated in neuronal cells following DNA damage. Evidence is presented to show that proper DNA damage response is critically important, not just for normal neuronal development but throughout the entire life of any organism. Defective DNA damage response in older human age can generate neurodegenerative disorders such as Alzheimer's or Parkinson diseases. PMID- 17115941 TI - The application of proteomics and genomics to the study of age-related neurodegeneration and neuroprotection. AB - The present study aimed to acquire more information on aging-related alterations, using proteomic and genomic analyses of hippocampus from young (8 months) and old (27 months) rats. In the old rats, the proteomic analysis identified changes in proteins related to the iron-mediated oxidative stress (OS) pathway, including reduction in antioxidant enzymes (e.g., peroxiredoxin, cytochrome c oxidase) and induction of ferritin. Furthermore, the neurofilament light peptide, associated with neurodegenerative processes, was enhanced and binding/ chaperone proteins were altered in old vs. young rats. At the genes levels, significant molecular changes related to neurodegeneration were identified in aged rat hippocampus. Thus, the effects of the potent neuroprotective compounds, the anti-Parkinson drug, rasagiline and the anti-Alzheimer drug, ladostigil (1 mg/kg, for 30 days) on gene expression in the hippocampus were further investigated. Both drugs reversed the effect of aging on the expression of various mitochondrial and key regulator genes involved in neurodegeneration, cell survival, synaptogenesis, oxidation, and metabolism. These results support the hypothesis that OS and mitochondrial dysfunction may play a pivotal role in aging and age-associated neurodegenerative diseases, and can serve as potential clinical targets for future therapy. PMID- 17115942 TI - Control of neuronal plasticity by reactive oxygen species. PMID- 17115943 TI - Does edaravone (MCI- 186) act as an antioxidant and a neuroprotector in experimental traumatic brain injury? AB - Edaravone (MCI-186) is a novel synthetic free radical scavenger intended to have neuroprotective effect against ischemic insult. It is currently used on patients with cerebral infarction. Here, we note beneficial pharmaceutical effects of edaravone in rat experimental traumatic brain injury. Under specific experimental conditions, edaravone minimized traumatic brain injury by functioning as a synthetic antioxidant. Clinical trials testing the efficacy of edaravone are warranted. PMID- 17115944 TI - H2O2 signaling in the nigrostriatal dopamine pathway via ATP-sensitive potassium channels: issues and answers. AB - The role of reactive oxygen species (ROS) as signaling agents is increasingly appreciated. Studies of ROS functions in the central nervous system, however, are only in their infancy. Using fast-scan cyclic voltammetry and fluorescence imaging in brain slices, the authors discovered that hydrogen peroxide (H2O2) is an endogenous regulator of dopamine release in the dorsal striatum. Given the key role of dopamine in motor, reward, and cognitive pathways, regulation by H2O2 has implications for normal dopamine function, as well as for dysfunction of dopamine transmission. In this review, data are summarized to show that H2O2 is a diffusible messenger in the striatum, generated downstream from glutamate receptor activation, and an intracellular signal in dopamine neurons of the substantia nigra, generated during normal pacemaker activity. The mechanism by which H2O2 inhibits dopamine release and dopamine cell activity is activation of ATP-sensitive K+ (KATP) channels. Characteristics of the neuronal and glial antioxidant networks required to permit H2O2 signaling, yet prevent oxidative damage, are also considered. Lastly, estimates of physiological H2O2 levels are discussed, and strengths and limitations of currently available methods for H2O2 detection, including fluorescence imaging using dichlorofluorescein (DCF) and the next generation of fluorescent probes, are considered. PMID- 17115945 TI - Phase I trial of intramuscular injection of a recombinant adeno-associated virus serotype 2 alphal-antitrypsin (AAT) vector in AAT-deficient adults. AB - A phase I trial of intramuscular injection of a recombinant adeno-associated virus serotype 2 (rAAV2) alpha1-antitrypsin (AAT) vector was performed in 12 AAT deficient adults, 10 of whom were male. All subjects were either homozygous for the most common AAT mutation (a missense mutation designated PI*Z) or compound heterozygous for PI*Z and another mutation known to cause disease. There were four dose cohorts, ranging from 2.1 x 10(12) vector genomes (VG) to 6.9 x 10(13) VG, with three subjects per cohort. Subjects were injected sequentially in a dose escalating fashion with a minimum of 14 days between patients. Subjects who had been receiving AAT protein replacement discontinued that therapy 28 days before vector administration. There were no vector-related serious adverse events in any of the 12 participants. Vector DNA sequences were detected in the blood between 1 and 3 days after injection in nearly all patients receiving doses of 6.9 x 10(12) VG or higher. Anti-AAV2 capsid antibodies were present and rose after vector injection, but no other immune responses were detected. One subject who had not been receiving protein replacement exhibited low-level expression of wild-type M AAT in the serum (82 nM), which was detectable 30 days after receiving an injection of 2.1 x 10(13) VG. Unfortunately, residual but declining M-AAT levels from the washout of the protein replacement elevated background levels sufficiently to obscure any possible vector expression in that range in most of the other individuals in the higher dose cohorts. PMID- 17115948 TI - Clinical scenarios. PMID- 17115949 TI - The Australian response: pandemic influenza preparedness. AB - Australia's preparedness for a potential influenza pandemic involves many players, from individual health carers to interdepartmental government committees. It embraces a wide number of strategies from the management of the disease to facilitating business continuity. The key strategy underlying Australia's planned response is an intensive effort to reduce transmission of the virus. This includes actions to reduce the likelihood of entry of the virus into the country and to contain outbreaks when they occur. Containment will provide time to allow production of a matched vaccine. The health strategies are outlined in the Australian health management plan for pandemic influenza. The plan is accompanied by technical annexes setting out key considerations and guidelines in the areas of clinical management and infection control. National plans present overall strategies and guidance, but the operational details can only be determined by individual states and territories, regions, and the services themselves. Primary health care practices will be on the frontline of an influenza pandemic. Every practice needs a plan that defines the roles of staff, incorporates infection control and staff protection measures, and considers business continuity. Most importantly, a practice needs to know how to implement that plan. PMID- 17115950 TI - The influenza viruses. AB - Human epidemic influenza is caused by influenza type A and B viruses, which continually undergo antigenic change in their surface antigens, haemagglutinin (H) and neuraminidase (N). Influenza epidemics are the consequence of small, ongoing antigenic changes known as "antigenic drift", which occurs in both influenza types. Pandemic influenza occurs at irregular and unpredictable intervals, and is the result of a major antigenic change known as "antigenic shift", which occurs only in influenza A. Aquatic birds are the evolutionary hosts of influenza viruses; they harbour many distinct forms or subtypes of influenza A, which are usually present as harmless gut infections. Antigenic shift involves the evolution of a new human influenza A virus through the acquisition of a new haemagglutinin gene encoding a different subtype from an avian influenza, or by the adaptation of an avian virus, causing it to become transmissible between humans. Two subtypes of avian influenza, H5 and H7, can cause severe infections when introduced into domestic poultry. Recently, influenza A/H5N1 viruses have caused widespread outbreaks, starting in Asia and spreading widely to other regions. Avian influenza viruses do not readily infect humans. However, during the past 3 years, more than 250 cases of H5N1 infection of humans have occurred, with associated mortality approaching 60%. It is feared that a new pandemic of human influenza may emerge from this. PMID- 17115951 TI - Pandemic influenza: clinical issues. AB - Influenza is an acute febrile illness caused by influenza A or B viruses. It occurs mainly in winter in temperate climates, and throughout the year in tropical Australia. It is highly contagious and of considerable public health concern because of the rapidity with which epidemics evolve and the associated morbidity and mortality. Most influenza illnesses resolve over about 1 week without specific medical intervention. People at particular risk for complicated infection are those > 65 or < 5 years old, those with chronic medical comorbidities, residents of chronic care facilities (including nursing homes), and women in the second or third trimester of pregnancy. Complicated influenza infection most commonly manifests as primary viral pneumonia, combined viral and bacterial pneumonia, and secondary bacterial pneumonia. Rare but serious complications of influenza include central nervous system involvement (eg, encephalitis, transverse myelitis, aseptic meningitis, and Guillain-Barre syndrome). The recent emergence of avian influenza A/H5N1 and confirmation of sporadic cases of human H5N1 infection have heightened concern about an impending human influenza pandemic, either from a human form of H5N1 or a primary new human influenza strain. H5N1 infection in humans has been associated with severe illness and a > 50% mortality rate, with high mortality in people aged 10-39 years. PMID- 17115952 TI - Laboratory diagnosis of human seasonal and pandemic influenza virus infection. AB - Laboratory diagnosis is important to distinguish influenza from other respiratory virus infections. It will be especially important in detecting the first cases of pandemic influenza. Good quality respiratory tract sampling is needed to maximise diagnostic yield in influenza infection. In the appropriate clinical setting, pandemic strain-specific nucleic acid testing is the initial test of choice for suspected pandemic influenza. It is more sensitive than virus isolation, and more sensitive and specific than serology, immunofluorescence and other antigen detection methods. Virus isolation is needed to monitor new influenza strains and for vaccine development. Analysis of influenza isolates is undertaken by the World Health Organization Global Influenza Surveillance Network. Monitoring for antiviral resistance will be needed with widespread use of neuraminidase inhibitors for treatment and prophylaxis during a pandemic. PMID- 17115953 TI - Infection control and pandemic influenza. AB - If an influenza pandemic occurs, the spread of the virus should be reduced for as long as possible while an effective vaccine is produced. Influenza spreads mainly by large respiratory droplets (> 5 microm) depositing onto the mucosal surfaces of the eye, mouth or respiratory tract. Hands are another major means for spread, and are frequently contaminated by droplets. The most effective way to reduce the spread of the virus is with good infection control practices and social distancing. Infection control practices include the use of personal protective equipment (PPE), hand hygiene, and respiratory hygiene and cough etiquette. Infected people should be isolated and spatial separation observed in common areas where infected people may be present. Any practices that create aerosols (eg, nebulisation) should be avoided, unless performed with appropriate precautions, especially with all people in the room wearing appropriate PPE. Now is the time to re-examine all our current practices so that we are better prepared, well practised and have good infection control practices in place for all transmissible respiratory infections. PMID- 17115954 TI - Antivirals in the management of an influenza pandemic. AB - The Australian Government has an extensive stockpile of antivirals (neuraminidase inhibitors) to be used if an influenza pandemic occurs. Neuraminidase inhibitors reduce the duration of the symptoms of seasonal influenza infection by 1 day on average, when used as treatment within 48 hours of disease onset. Neuraminidase inhibitors prevent infection in up to 74% of people when administered as prophylaxis. Resistance of seasonal influenza viruses to neuraminidase inhibitors is low. The safety and efficacy (including resistance) of neuraminidase inhibitors against pandemic influenza or the virus of current concern in pandemic planning, influenza A/H5N1, is not known, and further research is needed. PMID- 17115955 TI - Pandemic vaccines: promises and pitfalls. AB - Prototype vaccines against influenza A/H5N1 may be poorly immunogenic, and two or more doses may be required to induce levels of neutralising antibody that are deemed to be protective. The actual levels of antibody required to protect against a highly pathogenic virus that potentially can spread beyond the large airways is unknown. The global capacity for vaccine manufacture in eggs or tissue culture is considerable, but the number of doses that can theoretically be produced in a pandemic context will only be sufficient for a small fraction of the world's population, even less if a high antigen content is required. The safety of new pandemic vaccines should be addressed in an internationally coordinated way. Steps are underway through the Therapeutic Goods Administration to evaluate mock-up vaccines now, so that the time to registration of a new product can be minimised. It will be 3-6 months into the pandemic before an effective vaccine becomes available, so other control measures will be important in the early stages of a pandemic. The primary goal of a pandemic influenza vaccine must be to prevent death, and not necessarily to prevent infection. PMID- 17115956 TI - General practice: professional preparation for a pandemic. AB - General practice will play a key role in both prevention and management of an influenza pandemic. Australian pandemic plans acknowledge a role for general practice, but there are few published data addressing the issues that general practitioners and their practices will face in dealing with such a crisis. The outcome will revolve around preparation in three key areas: Definition of the role of general practice within a broad primary care pandemic response, and adequate preparation within general practices so they can play that role well. Planning exercises and forums must include GPs, and rehearsals must include practical experience for general practices and their staff. Local Divisions of General Practice and GP practices can advocate for this, can define their role, and can prepare by using pandemic preparedness checklists; Definition and enactment of communication strategies to facilitate transfer of useful clinical and administrative data from practices and rapid dissemination of information into the community via general practice; Resource provision, which should be centrally funded but locally distributed, with personal protective equipment, vaccines and antivirals readily available for distribution. Resources must include support for human resource management to ensure appropriate health care professionals reach areas of workforce demand. Administrative, clinical and financial resources must be available to train GPs and practices in pandemic awareness and response. PMID- 17115957 TI - Pandemic influenza and critical infrastructure dependencies: possible impact on hospitals. AB - Hospitals will be particularly challenged when pandemic influenza spreads. Within the health sector in general, existing pandemic plans focus on health interventions to control outbreaks. The critical relationship between the health sector and other sectors is not well understood and addressed. Hospitals depend on critical infrastructure external to the organisation itself. Existing plans do not adequately consider the complexity and interdependency of systems upon which hospitals rely. The failure of one such system can trigger a failure of another, causing cascading breakdowns. Health is only one of the many systems that struggle at maximum capacity during "normal" times, as current business models operate with no or minimal "excess" staff and have become irreducible operations. This makes interconnected systems highly vulnerable to acute disruptions, such as a pandemic. Companies use continuity plans and highly regulated business continuity management to overcome process interruptions. This methodology can be applied to hospitals to minimise the impact of a pandemic. PMID- 17115958 TI - Ethical issues in pandemic planning. AB - In the event of an influenza pandemic, many ethical issues will arise in terms of health risks, resource allocation, and management decisions. Planning decisions may be controversial, such as rationing of antivirals, resource allocation (including hospital beds and vaccinations), occupational risk, rostering of staff, responsibilities of health care workers, quarantine measures, and governance issues. A clear ethical framework is needed to enable understanding of the decision-making process and optimise acceptance of decisions by health care workers and other members of an affected community. Planning decisions need to start being examined now, and will require input from a broad group of experts: health care providers, infrastructure managers, lawyers, ethicists, public health physicians, and community members. The process will need to be open, honest and dynamic. PMID- 17115959 TI - Urgent strategic research into influenza to inform health policy and protect the public. AB - The Australian management plan for pandemic influenza (2005) highlighted a number of areas where more information may yield better plans for protecting Australia. In 2005, the National Health and Medical Research Council (NHMRC) developed a special "urgent research" funding program to meet those information needs as quickly as possible. The funding program resulted in grants totalling $6.5 million being awarded for 33 research projects, in five broad areas: Detection and identification of the virus; Vaccine development and evaluation; Antiviral medication use and effectiveness; Public health interventions; and Understanding behavioural responses to achieve effective communication and staged implementation of public health strategies. Outcomes of the program will be evaluated formally in 2007. PMID- 17115961 TI - Preserving the fertility of children with cancer. AB - Moving beyond the uncertainties of risk, and limited, often difficult, preservation options, will require consensus and collaborative research. PMID- 17115962 TI - Technologies for the diagnosis of primary melanoma of the skin. AB - There is now an inexpensive first-line approach for diagnosing pigmented skin lesions. PMID- 17115963 TI - Tackling partner violence in families. AB - New guidelines extend opportunities for GPs to respond. PMID- 17115964 TI - Fertility preservation in children newly diagnosed with cancer: existing standards of practice in Australia and New Zealand. AB - OBJECTIVE: To establish the extent to which sperm, oocyte and gonadal tissue collection and storage is offered to children newly diagnosed with cancer. DESIGN, PARTICIPANTS AND SETTING: A cross-sectional survey of all paediatric oncology services in Australia and New Zealand (ANZ) in December 2005. MAIN OUTCOME MEASURES: Sperm, oocyte and gonadal tissue collection and storage practices at paediatric oncology services; comparisons with recently published North American practices and with current recommendations for best practice. RESULTS: 12 of the 13 centres (92%) completed the survey. All centres offered sperm preservation, but only 10 (83%) offered oocyte/ovarian tissue preservation. Two centres were using gonadotrophin-releasing hormone analogues for fertility protection in postpubertal females. Five (42%) had offered fertility preservation to patients before the completion of their sexual development. All centres were more likely to offer sperm preservation than oocyte preservation for any given disease. The most common diseases for which conservation was offered were lymphomas and sarcomas. The anticipated cumulative dose at which centres elected to offer fertility preservation varied widely, both for the alkylator cyclophosphamide (any to 10 g/m(2)) and for abdominal/pelvic irradiation (any to 12 Gy) and spinal irradiation (any to 18 Gy). Fertility counselling was offered in a variety of settings by nine (75%) of the centres. Despite 11 centres (92%) agreeing that fertility preservation guidelines would be helpful, only two (17%) had guidelines in place. CONCLUSIONS: There are inconsistencies in the indications for and methods of gamete conservation in paediatric oncology centres throughout ANZ. Variations in practice on a background of unresolved medical, legal and ethical issues suggest the development of guidelines would be helpful. PMID- 17115965 TI - Dietary protein intakes in patients with hepatic encephalopathy and cirrhosis: current practice in NSW and ACT. AB - OBJECTIVE: To ascertain whether current practice in teaching hospitals in New South Wales and the Australian Capital Territory delivers adequate dietary protein in the management of malnutrition in adults with cirrhosis, in accordance with European Society for Clinical Nutrition and Metabolism (ESPEN) guidelines for nutrition in liver disease. STUDY DESIGN: Cross-sectional study of dietitians using a self-administered, mail-back survey. SETTING: Teaching hospitals in NSW and the ACT treating patients with cirrhosis. PARTICIPANTS: Dietitians seeing patients with cirrhosis in the 12 months prior to completing the survey. MAIN OUTCOME MEASURES: Current dietary protein prescription practice for patients with cirrhosis (with and without hepatic encephalopathy); use of nutritional supplements and enteral feeding for malnourished patients with cirrhosis. RESULTS: Dietitians following the ESPEN guidelines were in the minority: 36% of the dietitians recommended an adequate protein intake for patients with hepatic encephalopathy. Sixty-four per cent of the dietitians had received referrals from the medical team requesting inappropriate protein-restricted diets for patients without hepatic encephalopathy. Seventy-eight per cent of the dietitians requested clarification of the recommended nutritional management of patients with cirrhosis. CONCLUSION: Many medical and dietetic staff inappropriately restrict protein intake of patients with cirrhosis. PMID- 17115966 TI - Off-label use of medicines: consensus recommendations for evaluating appropriateness. AB - Off-label prescribing is the prescription of a registered medicine for a use that is not included in the product information. The practice is common, with rates up to 40% in adults and up to 90% in paediatric patients. Off-label prescribing is not illegal and may sometimes be clinically appropriate, but is associated with a number of clinical, safety and ethical issues. To date, no explicit guidance has been available to help clinicians assess appropriateness in off-label prescribing. We describe the development of a guide for clinicians, policymakers and funders of health care in evaluating the appropriateness of medicines proposed for off-label use. Three broad categories of appropriate off-label use are identified:off-label use justified by high-quality evidence; use within the context of a formal research proposal; and exceptional use, justified by individual clinical circumstances. An appropriate process for informed consent is proposed for each category. If there is no high-quality evidence supporting off label use, and the medicine is not suitable for exceptional or research indications, its use is generally not recommended. This will reduce inappropriate use, enhance patient safety by reducing exposure to unnecessary risk, and may stimulate more clinically relevant medicines research. PMID- 17115967 TI - Guidelines for the prevention, detection and management of people with chronic heart failure in Australia 2006. AB - Chronic heart failure (CHF) is found in 1.5%-2.0% of Australians. Considered rare in people aged less than 45 years, its prevalence increases to over 10% in people aged >/= 65 years. CHF is one of the most common reasons for hospital admission and general practitioner consultation in the elderly (>/= 70 years). Common causes of CHF are ischaemic heart disease (present in > 50% of new cases), hypertension (about two-thirds of cases) and idiopathic dilated cardiomyopathy (around 5%-10% of cases). Diagnosis is based on clinical features, chest x-ray and objective measurement of ventricular function (eg, echocardiography). Plasma levels of B-type natriuretic peptide (BNP) may have a role in diagnosis, primarily as a test for exclusion. Diagnosis may be strengthened by a beneficial clinical response to treatment(s) directed towards amelioration of symptoms. Management involves prevention, early detection, amelioration of disease progression, relief of symptoms, minimisation of exacerbations, and prolongation of survival. PMID- 17115968 TI - Wrongful life claims: dignity, disability and "a line in the sand". AB - A recent High Court decision held that children born with disabilities not caused by medical intervention, but not diagnosed antenatally, could not claim general damages for their pain and suffering, nor special damages for the needs created by their disabilities and their loss of earning capacity. PMID- 17115969 TI - Hendra virus infection in a veterinarian. AB - A veterinarian became infected with Hendra virus (HeV) after managing a terminally ill horse and performing a limited autopsy with inadequate precautions. Although she was initially only mildly ill, serological tests suggested latent HeV infection. Nevertheless, she remains well 2 years after her initial illness. Recently emerged zoonotic viruses, such as HeV, necessitate appropriate working procedures and personal protective equipment in veterinary practice. PMID- 17115970 TI - 6: Rhinitis and asthma: united airway disease. AB - United airway disease is characterised by inflammation of the respiratory tract, in which asthma and rhinitis are the upper and lower respiratory tract manifestations, respectively, of the same disease process. Irrespective of cause, the upper and lower respiratory tract manifestations are characterised by a systemic inflammatory response. Patients with rhinitis or asthma should always be assessed for coexistent disease in the reciprocal area. Treatment of upper airway disease can modify the severity of lower airway disease and vice versa. The potential for early treatment of allergic rhinitis to prevent progression to asthma merits further study. PMID- 17115971 TI - Allergy and sinus disease. PMID- 17115972 TI - Public reporting of hospital outcomes based on administrative data. PMID- 17115973 TI - Bill to ban reproduction of inmates with cancer proposed in New South Wales. PMID- 17115974 TI - More than task substitution and transfer. PMID- 17115975 TI - Bacillus minimum genome factory: effective utilization of microbial genome information. AB - In 1997, the complete genomic DNA sequence of Bacillus subtilis (4.2 Mbp) was determined and 4100 genes were identified [Kunst, Ogasawara, Moszer, Albertini, Alloni, Azevedo, Bertero, Bessieres, Bolotin, Borchert, S. et al. (1997) Nature 90, 249-256]. In addition, B. subtilis, which shows an excellent ability to secrete proteins (enzymes) and antibiotics in large quantities outside the cell, plays an important role in industrial and medical fields. It is necessary to clarify the genes involved in the production of compounds by understanding the network of these 4100 genes and the proceeding analysis of genes of unknown functions. In promoting such a study, it is expected that the regulatory system of B. subtilis can be simplified by the creation of a Bacillus strain with a reduced genome by discriminating genes unnecessary for the production of proteins from essential genes, and deleting as many of these unnecessary genes as possible, which may help to understand this complex network of genes. We have previously distinguished essential and non-essential genes by evaluating the growth and enzyme-producing properties of strains of B. subtilis in which about 3000 genes (except 271 essential genes) have been disrupted or deleted singly, and have successfully utilized the findings from these studies in creating the MG1M strain with an approx. 1 Mbp deletion by serially deleting 17 unnecessary regions from the genome. This strain showed slightly reduced growth in enzyme production medium, but no marked morphological changes. Moreover, we confirmed that the MG1M strain had cellulase and protease productivity comparable with that of the B. subtilis 168 strain, thus demonstrating that genome reduction does not contribute to a negative influence on enzyme productivity. PMID- 17115976 TI - Mechanical dissociation of human embryonic stem cell colonies by manual scraping after collagenase treatment is much more detrimental to cellular viability than is trypsinization with gentle pipetting. AB - Because hESC (human embryonic stem cells) are 'social cells' that require co operative interactions and intimate physical contact with each other, it is absolutely essential to dissociate hESC colonies into cellular clumps rather than into a single-cell suspension during serial passage. The present study compared two commonly used protocols for dissociating hESC colonies. The first protocol involved mild enzymatic treatment with collagenase type IV (1 mg/ml) for approx. 5-10 min, prior to mechanical dissociation into cellular clumps through manual scraping with a plastic pipette tip. The second protocol involved a short duration of exposure (2-3 min) to low concentrations of trypsin (0.05%), followed by gentle pipetting. The MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H tetrazolium bromide] assay was used to compare the recovery of viable cells after dissociating hESC colonies with these two protocols, before and after conventional freeze-thawing with 10% (v/v) DMSO. Besides undifferentiated hESC, the randomly differentiated fibroblastic progenies of hESC at various passages (P0-P4), together with an immortalized cell line (CRL-1486), were also utilized to compare the two protocols. The results demonstrated that the second protocol (trypsinization with gentle pipetting) is much less detrimental to cellular viability than is the first protocol (collagenase treatment with scratching). This in turn translated to higher freeze-thaw survival rates. It is hypothesized that scratching after collagenase treatment (first protocol) somehow induces physical damage to the cells, thereby leading to a lower recovery of viable cells, both before and after freeze-thawing. PMID- 17115977 TI - Cryptochromes impair phosphorylation of transcriptional activators in the clock: a general mechanism for circadian repression. AB - CLOCK and BMAL1 [brain and muscle ARNT (arylhydrocarbon receptor nuclear translocator)-like protein 1] are central components of the molecular clock in mammals and belong to the bHLH (basic helix-loop-helix)/PAS [PER (Period)/ARNT/SIM (single-minded)] family. Features of their dimerization have never been investigated. Here, we demonstrate that PAS domain function requires regions extending over the short PAS core repeats. Strikingly, while deleting PAS core repeats does not overtly affect dimerization, it abolishes the transcriptional activity of the heterodimer. Interestingly, these deletions also abolish co-dependent phosphorylation of CLOCK and BMAL1, suggesting a link between the phosphorylation status of the heterodimer and its transactivation potential. We demonstrate that NPAS2 (neuronal PAS domain protein 2) and BMAL2 also undergo similar posttranslational modifications, thereby establishing the mechanism proposed for CLOCK-BMAL1 as a common feature of transcriptional activators in the circadian clock. The discovery of two novel splice variants of BMAL2 confirms the crucial role of the PAS domain and further strengthens the view that co-dependent phosphorylation is of functional significance. In agreement with this, we demonstrate that CRY1-2 (cryptochromes 1-2) affect transactivation and phosphorylation of transcriptional activators of the clock. Furthermore, CRY proteins stabilize the unphosphorylated forms of BMAL1(BMAL2) thereby shifting the phosphorylated/unphosphorylated ratio towards a predominantly unphosphorylated (transcriptionally inactive) form. In contrast, PER proteins, which are weak repressors, are without effect. From these results, we propose a general mechanism for the inhibition of CLOCK(NPAS2)-BMAL1(BMAL2) circadian transcriptional activation by CRY1-2. PMID- 17115980 TI - A clinical study of migraine evolution. AB - BACKGROUND: The buildup time of migraine headaches has not been well delineated in publications to date and we are aware of patients whose migraines last well over 72 hours. More concentration on these factors in the assessment of patients might lead to more appropriate therapeutic choices. METHOD: Prospective ascertainment of such data through a questionnaire completed by 253 informed and willing patients with IHS migraine with or without aura consulting Canadian headache specialists. Data were electronically sent to a central computer from each center, tabulated and analyzed using standard statistical parameters. RESULTS: In 253 patients with migraine ascertained using applied IHS criteria, nausea was a feature in over 90% of cases, especially in those with aura. This inhibited the ingestion of oral medications in about a quarter of all subjects. The time to build from no pain to moderate/severe pain was shorter in subjects with auras and was less than 2 hours in 97% of those with and 86% of those without auras. However, we also identified a group of subjects with migraine (over 10% of all) in whom the build time to maximum pain is delayed for over 2 hours. Nausea was experienced by 91.7% of subjects, slightly but significantly later in those without auras. While most headaches in each group lasted from 4 to 72 hours, 24.3% of those with and 20.6% of those without aura expected to experience pain for more than 72 hours, while in untreated cases disability due to pain, nausea, or malaise usually persisted for over 3 days in 24.3% and 16.7% of those with and without aura, respectively. One-fifth of migraineurs may be in pain and/or disabled by accompanying symptoms for over 3 days in a typical migraine attack. Over half of our subjects reported that their medications worked well or excellently. CONCLUSIONS: Attacks of migraine in real-life clinical situations vary somewhat from the IHS criteria in that they are more often associated with nausea that interferes with oral therapy; can persist for over 72 hours; may have slow (>2 hours) buildup to maximum pain in 10% of cases; and may cause disability for over 3 days. Nevertheless, current therapeutic regimens (including prescribed medications) work well for a substantial majority. PMID- 17115981 TI - Prophylactic COX 2 inhibitor: an end to the Yom Kippur headache. AB - INTRODUCTION: Religious fasting is associated with headache. This has been documented as "Yom Kippur Headache" and "First of Ramadan Headache." The Cox2 inhibitor, rofecoxib, has been reported effective in preventing perimenstrual migraine and in preventing recurrence of migraine. Given its 17 hour half-life, we undertook this study to see whether 50 mg rofecoxib taken just prior to the 25 hour Yom Kippur fast would be effective in preventing headache. METHODS: We performed a double-blind randomized prospective trial of rofecoxib 50 mg versus placebo, taken just prior to the onset of fasting, Yom Kippur 2004. Healthy adults aged 18 to 65 were enrolled from the community and from hospital staff. Subjects completed a demographic data form and questions regarding headache history and a post-fast survey on headache during the fast, headache intensity, general ease of fasting, and side effects. RESULTS: We sent out 170 forms of which 105 were completed and returned. Of those subjects receiving rofecoxib (n = 53), 10 or 18.9% versus 34 or 65.4% of the placebo group (n = 52) had headache at some point during the fast (P < .0001). Severity of headache in the treatment group was significantly less for the treatment group (3.45 vs 6.29 on a visual analog scale of 10 (P= .009)). None of those receiving rofecoxib reported a "more difficult than usual fast" whereas the distribution of difficult to easy fast among the placebo group was more even. CONCLUSION: Rofecoxib 50 mg taken prior to a 25-hour ritual fast prevents and attenuates fasting headache. PMID- 17115982 TI - Effectiveness and tolerability of acupuncture compared with metoprolol in migraine prophylaxis. AB - OBJECTIVES: In a randomized controlled multicenter trial extending over 24 weeks, we investigated whether acupuncture is as effective and safe as metoprolol in the prophylactic treatment of migraine under conditions similar to routine care. METHODS: One hundred fourteen migraine patients could be randomized to treatment over 12 weeks either with acupuncture (8 to 15 sessions) or metoprolol (100 to 200 mg daily). Main outcome measure was the difference in the number of migraine days between baseline and the weeks 9 to 12 after randomization (derived from a headache diary). RESULTS: Two of 59 patients randomized to acupuncture withdrew prematurely from the study compared to 18 of 55 randomized to metoprolol. The number of migraine days decreased by 2.5 +/- 2.9 days (baseline 5.8 +/- 2.5 days) in the acupuncture group compared to 2.2 +/- 2.7 days (baseline 5.8 +/- 2.9 days) in the metoprolol group (P= .721). The proportion of responders (reduction of migraine attacks by > or =50%) was 61% for acupuncture and 49% for metoprolol. Both physicians and patients reported fewer adverse effects in the acupuncture group. CONCLUSIONS: Due to missing the recruitment target (480 patients) and the high drop-out in the metoprolol group the results must be interpreted with caution. Still, they suggest that acupuncture might be an effective and safe treatment option for patients unwilling or unable to use drug prophylaxis. PMID- 17115983 TI - Topiramate for migraine prevention in adolescents: a pooled analysis of efficacy and safety. AB - OBJECTIVE: To characterize the efficacy and safety of topiramate for migraine prevention in adolescents from 3 randomized, 26-week, double-blind, placebo controlled trials. BACKGROUND: Limited information is available regarding the efficacy and safety of prophylactic medications for treatment of adolescent migraine, a significant health problem. In studies that included adults and children, topiramate 100 and 200 mg/day were effective and generally well tolerated in the prevention of migraine headache. METHODS: We performed a post hoc subset analysis of the efficacy and safety data from the 51 patients, ages 12 17 years, enrolled in 3 pivotal trials of topiramate for migraine prophylaxis. RESULTS: Daily treatment with topiramate 50, 100, and 200 mg for 26 weeks reduced monthly migraine frequency from baseline 46% (P= .07), 63% (P= .02), and 65% (P= .04), respectively, compared with placebo (16%). Similarly, topiramate reduced both the monthly mean number of migraine days (1, 4, and 5 days for topiramate 50, 100, and 200 mg/day, respectively, vs 1 day for placebo) and percentage of days during which acute migraine medications were administered (59%, 54%, and 67% for topiramate 50, 100, and 200 mg/day, respectively, vs 42% for placebo), although the treatment differences did not reach nominal statistical significance. Topiramate 200 mg/day did not appear to offer greater efficacy than 100 mg/day. Treatment was generally well-tolerated, although adverse events were most frequent in the 200 mg/day dose group. CONCLUSIONS: This post hoc subset analysis suggests that topiramate 100 and 200 mg/day, and possibly 50 mg/day, administered prophylactically for 26 weeks may reduce migraine in adolescents. PMID- 17115984 TI - A novel method of eliciting pain-related potentials by transcutaneous electrical stimulation. AB - BACKGROUND: Electrophysiological techniques such as laser and contact heat evoked pain-related potentials are very useful for studying trigeminal and somatic pain transmission in humans. These methods are, however, partly invasive, expensive, and therefore not available for broad clinical use. We recently proposed a novel technique of noninvasive transcutaneous electrical stimulation. OBJECTIVE: To elicit pain-related evoked potentials (PREP) by using a concentric planar electrode and demonstrate their nociceptive specificity. METHODS: We registered PREP following stimulation of the forehead and hand in 14 healthy volunteers. Latencies, peak-to-peak amplitudes, and conduction velocities of nociceptive fibers have been estimated. Effects of temporal and spatial summation and of cutaneous anesthesia were evaluated. RESULTS: Stimulation with the concentric planar electrode produced pinprick-like painful sensation. Cutaneous anesthesia led to abolishment of PREP responses. Estimated mean conduction velocity was 11.61 +/- 5.12 m/s, which corresponded well with conduction via A-delta fibers. Spatial as well as temporal summation resulted in a parallel increase of perceived pain intensity and PREP amplitudes. CONCLUSION: The technique is noninvasive, affordable, and easy to perform and allows quantitative assessment of human nociceptive pathways. PMID- 17115985 TI - Gene expression profiling in cluster headache: a pilot microarray study. AB - BACKGROUND: Cluster headache (CH) is a primary neurovascular headache disorder characterized by attacks of excruciating pain accompanied by ipsilateral autonomic symptoms. CH pathophysiology is presumed to involve an activation of hypothalamic and trigeminovascular systems, but inflammation and immunological mechanisms have also been hypothesized to be of importance. OBJECTIVE: To identify differentially expressed genes during different clinical phases of CH, assuming that changes of pathophysiological importance would also be seen in peripheral venous blood. METHODS: Blood samples were drawn at 3 consecutive occasions from 3 episodic CH patients: during attacks, between attacks and in remission, and at 1 occasion from 3 matched controls. Global gene expression was analyzed with microarray tehnology using the Affymetrix Human Genome U133 2.0 Plus GeneChip Set, covering more than 54,000 gene transcripts, corresponding to almost 22,000 genes. Quantitative RT-PCR on S100P gene expression was analyzed in 6 patients and 14 controls. RESULTS: Overall, quite small differences were seen intraindividually and large differences interindividually. However, pairwise comparisons of signal values showed upregulation of several S100 calcium binding proteins; S100A8 (calgranulin A), S100A12 (calgranulin C), and S100P during active phase of the disease compared to remission. Also, annexin A3 (calcium binding) and ICAM3 showed upregulation. BIRC1 (neuronal apoptosis inhibitory protein), CREB5, HLA-DQA1, and HLA-DQB1 were upregulated in patients compared to controls. The upregulation of S100P during attack versus remission was confirmed by quantitative RT-PCR analysis. CONCLUSIONS: The S100A8 and S100A12 proteins are considered markers of non-infectious inflammatory disease, while the function of S100P is still largely unknown. Furthermore, upregulation of HLA-DQ genes in CH patients may also indicate an inflammatory response. Upregulation of these pro inflammatory genes during the active phase of CH has not formerly been reported. Data from this pilot microarray study provide a basis for further studies in CH. PMID- 17115986 TI - Change of excitability in brainstem and cortical visual processing in migraine exhibiting allodynia. AB - BACKGROUND: Clinical and neurophysiological manifestations of information processing associated with central sensitization are little known. Allodynic migraine (AM) can be caused by the sensitization of trigeminal neuron, but no study has reported on AM between attacks using blink reflex (BR) and pattern reversal visual evoked potentials (PVEPs). OBJECTIVE: We explored the characteristics of AM between attacks associated with central sensitization using BR and PVEP. METHODS: We recruited 13 patients with interictal AM and 15 patients with nonallodynic migraine (NA), and 30 healthy subjects (HS). BRs were obtained using paired pulses delivered at the interstimulus interval (ISI) of 150, 300, and 500 ms. The ratio of the area in the R2 of the second to R2 of the first shock was measured for each ISI. PVEP were recorded with 2 spatial frequencies (0.5 and 4.0 cpd) and 2 low and high contrasts (29% and 98%, respectively). Amplitudes of P100 were measured. RESULTS: For BR, there were no significant differences in the ratio of the area of the R2 between the sides of stimulation, and the sides of headache. AM patients had less suppression of the R2 at the ISI of 150 and 300 ms when compared with the NA patients and HS. For PVEP, at 0.5, there were significant differences of amplitude between AM patients and HS, and between NA patients and HS in low and high contrast. At 4.0 cpd, there were significant differences of amplitude between AM patients and HS in low contrast, and between AM patients and HS, and NA patients and HS in high contrast. In AM patients, there was a significant difference of amplitude ratio between 0.5 and 4.0 cpd. Conclusions.-Our BR and PVEP study showed that migraine patients exhibiting allodynia may show central sensitization of brainstem trigeminal neuron and have contrast modulating dysfunction during the cortical visual processing of striate and extrastriate on visual cortex in-between attacks. PMID- 17115987 TI - Effect of hyperoxia on neurogenic plasma protein extravasation in the rat dura mater. AB - OBJECTIVE: We aimed to study the effect of normobaric hyperoxia on neurogenic inflammation of the rat dura mater. BACKGROUND: Inhalation of 100% oxygen is a first-line therapy for the treatment of acute cluster headache (CH). However, the mechanisms underlying the antinociceptive effect of oxygen are poorly understood. Sumatriptan, which is also effective in aborting CH attacks, is known to inhibit neurogenic inflammation of the dura mater. We hypothesized that hyperoxia reduces dural plasma protein extravasation in the model of electrically stimulating the rat trigeminal ganglion. METHODS: Unilateral stimulation of the trigeminal ganglion was performed in anesthetized male Sprague-Dawley rats. We assessed plasma protein extravasation (PPE) in the ipsilateral dura mater under normoxic (group 1) and hyperoxic conditions (group 2: pO(2) 200 mmHg; group 3: pO(2) 300 mmHg; group 4: pO(2) 400 mmHg). The study results were compared to the effect of sumatriptan (300 microg/kg) on dural PPE. RESULTS: Under normoxic conditions, the calculated extravasation ratio was 1.72 +/- 0.2. Hyperoxic treatment (groups 2, 3, 4) significantly attenuated dural PPE. At oxygen levels of 400 mmHg, the PPE ratio was 1.14 +/- 0.2 (P < .01). After IV application of sumatriptan (300 microg/kg), PPE was nearly abolished (PPE ratio: 1.06 +/-0.17). CONCLUSION: Our findings demonstrate that hyperoxia is able to inhibit dural PPE. Hyperoxia may play an anti-inflammatory role in neurogenic inflammation, but further studies are needed to clarify whether this effect is either caused by prejunctional mechanisms or by modulation of the vascular permeability at postcapillary venules. PMID- 17115988 TI - The prophylactic treatment of chronic daily headache. AB - Chronic daily headache (CDH), a heterogeneous group of headache disorders occurring on at least 15 days per month, affects up to 4% to 5% of the general population. CDH disorders include transformed (or chronic) migraine, chronic tension-type headache, new daily persistent headache, and hemicrania continua. Patients with CDH have greater disability and lower quality of life than episodic migraine patients and often overuse headache pain medications. To date, only topiramate, gabapentin, tizanidine, fluoxetine, amitriptyline, and botulinum toxin type A (BoNTA) have been evaluated as prophylactic treatment of CDH in randomized, double-blind, placebo-controlled, or active comparator-controlled trials. The evidence supporting the use of BoNTA as prophylaxis of CDH is composed of larger and longer trials, as over 1000 patients were evaluated for up to 11 months duration. Compared with placebo BoNTA has significantly reduced the frequency of headache episodes, a recommended efficacy measure for headache trials and has been demonstrated to be safe and very well tolerated with few discontinuations due to adverse events. Side effects are generally transient, mild to moderate, and nonsystemic. The results of clinical trials using traditional oral pharmacotherapy, while supportive of their use as prophylactic treatment of CDH, are limited by several factors, including small numbers of patients, the choice of efficacy measures, and short treatment periods. The use of oral agents was associated with systemic side effects, which may limit their effectiveness as prophylactic treatment of CDH. PMID- 17115989 TI - Coexistence of TACS and trigeminal neuralgia: pathophysiological conjectures. AB - BACKGROUND: Trigeminal autonomic cephalgias (TACs) and trigeminal neuralgia are short-lasting unilateral primary headaches whose study is providing insights into craniofacial pain mechanisms. We report on 2 patients in whom trigeminal neuralgia coexists with the TACs paroxysmal hemicrania and SUNCT. CONCLUSION: Coexistence of trigeminal neuralgia with various TAC forms suggests a pathophysiological relationship between these short-lasting unilateral headaches. PMID- 17115990 TI - Circadian and seasonal variation of migraine attacks in children. AB - OBJECT: To investigate the rhythmicity of migraine episodes without aura in a pediatric population. METHODS: Time of occurrence of 2517 migraine attacks in 115 children was recorded, by means of a diary, both by hourly and monthly intervals. RESULTS: A significant circadian variation, characterized by a peak in the afternoon (P < .001) and one in the early morning (P= .002) was found. A seasonal peak was also observed between November and January, while a nadir was observed in July. CONCLUSIONS: The clustering of attacks in the morning and midday and in autumn-winter, with a minimum frequency in July, suggests that school activities may represent an important cause of migraine. PMID- 17115991 TI - Endothelial nitric oxide synthase (Glu298Asp) polymorphism is an independent risk factor for migraine with aura. AB - OBJECTIVE: The aim of the present study was to evaluate whether the functional endothelial nitric oxide synthase (eNOS) Glu298Asp polymorphism, which has been demonstrated to decrease the endothelial NOS activity, might be a risk factor for migraine. BACKGROUND: It has widely demonstrated that nitric oxide (NO) is involved in migraine pathogenesis. Several genetic risk factors have been associated with migraine, but no study has unraveled a possible relationship between migraine and eNOS Glu298Asp. Methods.-One hundred fifty-six migraine patients and 125 healthy nonheadache volunteers entered the study. Demographic and clinical characteristics were carefully recorded, and a neurological workup was performed. RESULTS: eNOS AspAsp homozygous patients had a 3-fold time risk for migraine with aura (MA) when compared to migraine without aura (MO) patients (OR-3.02, 95% CI-1.21 to 7.51), and more than 2-fold time increased risk when compared to control subjects (OR-2.21, 95% CI-1.00 to 5.04). In migraine patients, no difference in age at onset, mean attack's intensity, family history for any of the studied comorbidities, or the presence of comorbidities was found in eNOS AspAsp homozygous compared to eNOS GluGlu or eNOS GluAs carriers. CONCLUSIONS: Homozygous Asp298, a common variant of the eNOS gene, is an independent risk factor for MA in this study population. PMID- 17115992 TI - Headaches associated with Rathke's cleft cyst. AB - OBJECTIVE: Headaches are common presentations in patients with Rathke's cleft cyst (RCC). This study was conducted to elucidate the characteristics and underlying mechanisms of the headache. METHODS: We retrospectively studied 46 patients with RCC, 33 of whom underwent surgical intervention. RESULTS: Headache, particularly frontal headache, was the most common symptom, occurring in 27 patients (58.7%). Eleven patients with sudden episodic headache mimicking that of pituitary apoplexy. Although the presence of headache did not correlate with cyst size, it was significantly more common in RCCs with: high- and iso intensity content on T1-weighted MR image (P= .0363), mucous content within the cyst (P= .0023), and intense chronic inflammation at the cyst wall (P= .0276). Among 6 patients with histologically recognized intense inflammation, every patient had frontal headache (P= .0407), 5 patients had episodic headache (P= .0002), and 4 patients had associated hypopituitarism (P= .0073), none of which improved after surgical intervention. Headaches improved after surgery in 17 of 21 patients (81.0%). CONCLUSIONS: Headache, particularly frontal episodic headache, is a common and characteristic manifestation in patients with RCC and may indicate intermittent inflammatory reactions caused by mucous content. Patients with episodic headache should undergo surgical treatment to prevent exacerbation of the inflammation that can result in irreversible endocrine dysfunction. PMID- 17115993 TI - Pituitary tumor and apoplexy. PMID- 17115994 TI - Adult-onset ophthalmoplegic migraine with recurrent sixth nerve palsy: a case report. PMID- 17115995 TI - Occipital neuralgia evoked by facial herpes zoster infection. AB - Occipital neuralgia is a pain syndrome which may usually be induced by spasms of the cervical muscles or trauma to the greater or lesser occipital nerves. We report a patient with occipital neuralgia followed by facial herpes lesion. A 74 year-old male experienced sudden-onset severe headache in the occipital area. The pain was localized to the distribution of the right side of the greater occipital nerve, and palpation of the right greater occipital nerve reproduces the pain. He was diagnosed with occipital neuralgia according to ICHD-II criteria. A few days later, the occipital pain was followed by reddening of the skin and the appearance, of varying size, of vesicles on the right side of his face (the maxillary nerve and the mandibular nerve region). This was diagnosed as herpes zoster. This case represents a combination of facial herpes lesions and pain in the C2 and C3 regions. The pain syndromes can be confusing, and the classic herpes zoster infection should be considered even when no skin lesions are established. PMID- 17115996 TI - Noise as a trigger for headaches? A comment. PMID- 17115997 TI - Transient global amnesia and migraine. PMID- 17116000 TI - Crossing the barrier: oxysterols as cholesterol transporters and metabolic modulators in the brain. AB - A normal brain function requires constant levels of cholesterol, and the need for constancy seems to be higher here than in any other organ. Nature has met this need by isolation of brain cholesterol by a highly efficient blood-brain barrier. As a low synthesis of cholesterol is present in the brain, a mechanism for compensatory elimination is required. A decade ago we made the unexpected finding that the favoured mechanism for this involves conversion into 24S hydroxycholesterol, followed by diffusion over the blood-brain barrier. Recent studies by us and others on this new pathway have given new insights into the mechanisms by which cholesterol homeostasis is maintained in the brain. We recently demonstrated a flux of another oxygenated product of cholesterol, 27 hydroxycholesterol, in the opposite direction. The latter flux may be important for neurodegeneration, and may be the link between hypercholesterolaemia and Alzheimer's disease. An overview of the above studies is presented and the possibility that the cholesterol 24S-hydroxylase in the brain may be important for memory and learning and that it may be a new drug target is discussed. PMID- 17116001 TI - Intravenous polyclonal IgM-enriched immunoglobulin therapy in sepsis: a review of clinical efficacy in relation to microbiological aetiology and severity of sepsis. AB - The efficacy of intravenous polyclonal immunoglobulin (IVIG) as adjunct therapy in sepsis has long been debated. Clinical trials have yielded contradicting results, in part due to the varying study design and varying microbiological aetiologies. In most trials, the study drug has been IVIG containing polyclonal IgG. However, in recent reports, the efficacy of IgM-enriched IVIG as adjunct therapy in sepsis has been highlighted. Here we review studies on IgM-enriched IVIG therapy in sepsis and we discuss the clinical efficacy in relation to microbiological aetiology and severity of sepsis. The results suggest that patients most likely to benefit from IgM-enriched IVIG therapy are those with Gram-negative septic shock. PMID- 17116002 TI - The use of vascular biomarkers and imaging studies in the early clinical development of anti-tumour agents targeting angiogenesis. AB - Recent findings about the pathomechanisms of tumour angiogenesis have led to new therapeutic options in the treatment of malignant tumours. During the development of anti-angiogenic drugs, reporting ranged from healing cancer to completely ineffective drugs. In 2003 the first anti-angiogenic drug was approved. Several anti-angiogenic drugs are still in the clinical phase of development. In contrast to identifying the maximal tolerable dose, determination of the optimal biological dose--reaching biological activity at lower doses--has become the main target in the early development of anti-angiogenic agents. This has been evaluated by different biomarker techniques. As a new standard in anti-tumour treatment, a better understanding of imaging in the treatment monitoring for anti angiogenic agents is important. Studies of tumour angiogenesis by tissue sampling rely on invasive procedures, adequate sampling and painstaking estimation of histological microvessel density. Attempts to develop wound healing assays to correlate angiogenesis in wounds with angiogenesis in tumour have been made but are still considered invasive and correlation of healthy with malignant tissue is still of limited validity. Several soluble markers of tumour angiogenesis were detected in various malignant diseases and were evaluated for assessing their use as surrogate markers in tumour angiogenesis. Further, soluble markers were investigated for visualizing them as imaging tools. Combining both, new soluble markers and imaging techniques, developing anticancer drugs and monitoring of therapy success becomes a dynamic process in which finally the patients' individual response can be achieved soon. Time-consuming delays for anatomically based restaging procedures can be avoided. Characterization of soluble biomarkers as well as different imaging techniques such as ultrasound, computed tomography (CT), magnetic resonance imaging and positron emission tomography combined with or without CT are reviewed in this manuscript. PMID- 17116003 TI - Circulating intestinal fibroblast growth factor 19 has a pronounced diurnal variation and modulates hepatic bile acid synthesis in man. AB - Bile acids (BAs) traversing the enterohepatic circulation exert several important metabolic effects. Their hepatic synthesis, controlled by the enzyme cholesterol 7alpha-hydroxylase (CYP7A1), has a unique diurnal variation in man. Here we provide evidence that the transintestinal flux of BAs regulates serum levels of intestinal fibroblast growth factor 19 (FGF19) that in turn modulate BA production in human liver. Basal FGF19 levels varied by 10-fold in normal subjects, and were reduced following treatment with a BA-binding resin and increased upon feeding the BA chenodeoxycholic acid. Serum FGF19 levels exhibited a pronounced diurnal rhythm with peaks occurring 90-120 min after the postprandial rise in serum BAs. The FGF19 peaks in turn preceded the declining phase of BA synthesis. The diurnal rhythm of serum FGF19 was abolished upon fasting. We conclude that, in humans, circulating FGF19 has a diurnal rhythm controlled by the transintestinal BA flux, and that FGF19 modulates hepatic BA synthesis. Through its systemic effects, circulating FGF19 may also mediate other known BA-dependent effects on lipid and carbohydrate metabolism. PMID- 17116004 TI - Prognostic implications of renal dysfunction in patients with stable angina pectoris. AB - OBJECTIVE: Impaired renal function is emerging as an independent risk factor for cardiovascular (CV) disease. We analysed the prognostic implications of estimated renal function in patients with angina pectoris. DESIGN: Post hoc analysis of the Angina Prognosis Study In Stockholm (APSIS). The estimated creatinine clearance (eCrCl) was calculated according to the Cockcroft-Gault formula in 808 patients. Outcomes were compared for subgroups with CrCl > or =90, 60-89 and<60 mL min(-1). Setting. Hospital-based study with patients referred from primary care and hospital. SUBJECTS: A total of 809 patients (248 women) with clinically diagnosed stable angina pectoris. Intervention. Double-blind treatment with metoprolol or verapamil. RESULTS: One hundred and sixty-four patients (91 women) had an eCrCl below 60 mL min(-1). During a median follow-up of 40 months, 38 patients suffered CV death and 31 patients had a nonfatal myocardial infarction (MI). In a univariate analysis a lower eCrCl was related to a higher risk for CV death or MI amongst men (log rank P = 0.036). A multivariate Cox analysis showed an independent prognostic importance of eCrCl for CV death (P = 0.046) and for CV death or MI (P = 0.042) amongst all patients. When analysed as a continuous variable, a 1 mL min(-1) decrease in eCrCl was associated with a 1.6% (0.1-3.1) increase in the risk for CV death or MI, and a 2.1% (0-4.1) increase in the risk for CV death alone. CONCLUSION: Renal dysfunction was found to be common in patients with stable angina pectoris and estimated creatinine clearances carried significant independent prognostic information regarding CV death and nonfatal MI. PMID- 17116005 TI - Use of integrated FDG PET/CT imaging in pulmonary carcinoid tumours. AB - BACKGROUND: Integrated positron emission tomography (PET)/computed tomography (CT) scanners have been recently introduced in the diagnostic work-up of suspected pulmonary malignancy and demonstrate encouraging results in the staging of nonsmall-cell lung cancer. OBJECTIVE: To evaluate the usefulness of integrated FDG PET/CT in pulmonary carcinoid tumours. SETTING: University hospital. METHODS: We studied 13 patients (mean age +/- 1 SD, 57 +/- 11 years) with pulmonary carcinoid tumours. All patients demonstrated a single pulmonary lesion. Integrated PET/CT scan and surgical resection were performed in all patients. RESULTS: The pulmonary lesion size ranged from 1.1 to 5.0 cm. Final histological diagnosis confirmed 12 typical and one atypical pulmonary carcinoid. Mean proliferation rate of the typical carcinoids was 1.7 +/- 1.4%. None of the patients had recurrent carcinoid disease or died during follow-up (864 +/- 218 days). Mean standardized uptake value (SUV) of (18)F-fluorodeoxyglucose (FDG) in typical carcinoids was 3.0 +/- 1.5 (range 1.2 - 6.6); SUV in the atypical carcinoid was remarkably high with a value of 8.5. The SUV was lower than 2.5 in 6 of 12 patients (50%). Mediastinal lymph node metastases or extrathoracic metastases were not detected in any patient. CONCLUSIONS: (18)F fluorodeoxyglucose PET/CT imaging improves accurate localization of metabolic activity and thus the interpretation of pulmonary lesions on CT. FDG uptake in pulmonary carcinoid tumours is often lower than expected for malignant tumours. Therefore, surgical resection or biopsy of lesions suspected to be carcinoids should be mandatory, even if they show no hypermetabolism on FDG PET images. PMID- 17116006 TI - Time trends in population cholesterol levels 1986-2004: influence of lipid lowering drugs, obesity, smoking and educational level. The northern Sweden MONICA study. AB - OBJECTIVES: To explore time trends in population total cholesterol. DESIGN AND SETTING: Five population-based cross-sectional surveys, 1986-2004 in the northern Sweden MONICA study included 8827 men and women. RESULTS: Age-adjusted cholesterol level declined in men, 25-64 years old, from 6.38 to 5.78 mmol L(-1) and in women from 6.32 to 5.51 mmol L(-1). Between 1994 and 2004, subjects 65-74 years old were included, and their levels also decreased, in men from 6.35 to 5.76 mmol L(-1) and in women from 7.11 to 6.24 mmol L(-1). The decrease was continuous over surveys and age groups, except in young and middle-aged men where no further decline was found after 1999. Cohorts born 1920-1939 showed decreased cholesterol over the period, whilst no change was noted for those born thereafter. In 2004, one-fourth of men and one-third of women 25-74 years achieved levels below 5.0 mmol L(-1). Subjects with low educational level, body mass index > or =25 or smokers all had higher cholesterol levels which persisted during the 18-year period. In 2004, the 9% who used lipid-lowering drugs are estimated to contribute, at most, to 0.13 mmol L(-1) lower cholesterol in the population. CONCLUSION: Large decreases in cholesterol levels occurred in the 18 year period. Less smoking may contribute to, and increasing obesity attenuate, this trend whilst lipid-lowering drugs have had little effect until recently. Socio-economic inequalities persist. PMID- 17116007 TI - Reduced lung function predicts increased fatality in future cardiac events. A population-based study. AB - OBJECTIVE: Moderately reduced lung function in apparently healthy subjects has been associated with incidence of coronary events. However, whether lung function is related to the fatality of the future events is unknown. This study explored whether reduced forced vital capacity (FVC) and forced expiratory volume in 1 s (FEV(1)) in initially healthy men is related to the fatality of the future coronary events. DESIGN: Prospective cohort study. SETTING: Population-based study from Malmo, Sweden. SUBJECTS: A total of 5452 healthy men, 28-61 years of age. MAIN OUTCOME MEASURES: Incidence of first coronary events was monitored over a mean follow-up of 19 years. The fatality of the future events was studied in relation to FEV and FVC. RESULTS: A total of 589 men suffered a coronary event during follow-up, 165 of them were fatal during the first day. After risk factors adjustment, low FEV or FVC were associated with incidence of coronary events (fatal or nonfatal) and this relationship was most pronounced for the fatal events. Amongst men who subsequently had a coronary event, the case-fatality rates were higher in men with low FEV or FVC. Adjusted for risk factors, the odds ratio for death during the first day was 1.00 (reference), 1.63 (95% CI: 0.9 3.1), 1.86 (1.0-3.5) and 2.06 (1.1-3.9), respectively, for men with FVC in the 4th, 3rd, 2nd, and lowest quartiles (trend: P < 0.05). FEV showed similar relationships with the fatality rates. CONCLUSION: Apparently healthy men with moderately reduced lung function have higher fatality in future coronary events, with a higher proportion of coronary heart disease deaths and less nonfatal myocardial infarction. PMID- 17116008 TI - Circulating levels of pro-atrial natriuretic peptide in lower respiratory tract infections. AB - OBJECTIVE: To analyse the mid region of plasma N-terminal pro-atrial natriuretic peptide (MR-proANP) levels in patients with lower respiratory tract infections to evaluate its prognostic use for the severity of disease and outcome. DESIGN: Prospective observational study. Setting. Emergency department of a university hospital. SUBJECTS: A total of 545 consecutive patients with lower respiratory tract infections and 50 healthy controls. Interventions. MR-proANP was measured in serum from all patients using a new sandwich immunoassay. RESULTS: MR-proANP levels (median [IQR], in pmol L(-1)) were significantly higher in patients with lower respiratory tract infections when compared with controls (138.0 [74.1 279.0] vs. 72.7 [62.5-89.5], P < 0.001), with highest levels in patients with community-acquired pneumonia (CAP). MR-proANP, but not C-reactive protein (CRP) levels, gradually increased with increasing severity of CAP, classified according to the pneumonia severity index (PSI) score (P < 0.001). On admission, MR-proANP levels were significantly higher in nonsurvivors when compared with survivors (293.0 [154.0-633.0] vs. 129.0 [71.4-255.0], P < 0.001). In a receiver operating characteristic (ROC) analysis for the prediction of survival of patients with CAP the area under the ROC curve (AUC) for MR-proANP was 0.69, similar when compared with the PSI (AUC 0.74, P = 0.31), and better when compared with other biomarkers, i.e. procalcitonin (AUC 0.57, P = 0.08), CRP (AUC 0.52, P = 0.02), and leucocyte count (AUC 0.56, P = 0.07). CONCLUSIONS: MR-proANP levels are increased in lower respiratory tract infections, especially in CAP. Together with other clinical, radiographic and laboratory findings, MR-proANP levels might be helpful for the risk stratification in CAP. PMID- 17116009 TI - An open-label study of darbepoetin alfa administered once monthly for the maintenance of haemoglobin concentrations in patients with chronic kidney disease not receiving dialysis. AB - OBJECTIVE: To demonstrate the efficacy and safety of once-monthly (QM) darbepoetin alfa administration in maintaining haemoglobin (Hb) 11.0-13.0 g dL( 1) in subjects with chronic kidney disease (CKD) not receiving dialysis and previously treated with darbepoetin alfa every other week (Q2W). SUBJECTS: This open-label study enrolled subjects > or =18 years of age who had glomerular filtration rate > or =15 and < or =60 mL min(-1)/1.73 m(2), had Hb 11.0-13.0 g dL(-1), and were receiving Q2W darbepoetin alfa. DESIGN: Subjects were switched to QM darbepoetin alfa therapy for 28 weeks; the QM dose was titrated to maintain Hb levels. Primary end-point: proportion of subjects maintaining Hb > or =11.0 g dL(-1) during the final 8 weeks of the study (evaluation phase). Secondary end points: Hb concentration during evaluation, darbepoetin alfa dose during the study, adverse events, laboratory parameters, and blood pressure. RESULTS: The study enrolled 152 subjects (female 52%, white 64%). Mean Hb > or =11.0 g dL(-1) during evaluation was achieved by 76% of the 150 subjects who received at least one dose of darbepoetin alfa [95% confidence interval (CI): 68%, 83%]. Mean (SD) Hb during evaluation was 11.71 (0.92) g dL(-1). Eighty-five per cent of 129 subjects who completed the study (95% CI: 78%, 91%) had Hb > or =11.0 g dL(-1) during evaluation. The dose of darbepoetin alfa over the study period was median (95% CI) 124.4 mug (106.2, 140.0). Darbepoetin alpha administered QM was well tolerated in study subjects. CONCLUSION: Darbepoetin alpha administered QM maintained Hb in study subjects with CKD not receiving dialysis. PMID- 17116010 TI - The effect of emergency department delay on outcome in critically ill medical patients: evaluation using hospital mortality and quality of life at 6 months. AB - OBJECTIVE: To assess the impact of delay in emergency department (ED) on outcome of critically ill patients admitted to the medical intensive care unit (MICU). Outcome was defined as hospital mortality and as health-related quality of life (HRQoL) at 6 months after intensive care assessed by the 15D measure. The 15D is a generic, 15-dimensional, standardized measure of HRQoL. We hypothesized that prolonged stay in the ED is related to worse outcome. DESIGN AND SETTING: A prospective follow-up cohort study in university hospital. SUBJECTS: All consecutive 1675 patients admitted to the MICU between July 2002 and June 2004. RESULTS: The 15D questionnaire was mailed to all patients alive at 6 months after admission. Of all MICU patients, 64% were admitted from ED. The mean length of stay in the ED was 6.2 h (95%CI 5.9-6.5 h). The hospital mortality rate was 24.4% (20.0% in the ED vs. 33.0% in the non-ED cohort, P < 0.001) and it was associated with higher age and degree of physiological derangement at admission. Neither the length of ED stay was associated with hospital mortality (P = 0.82) nor with HRQoL at 6 months after MICU admission (P = 0.34). Altogether, HRQoL at 6 months was significantly lower compared with the age- and sex-matched general population (P < 0.001). CONCLUSIONS: In a university hospital, the length of ED stay was not associated with the outcome of critically ill medical patients. However, we feel that the effect of ED treatment and delay on outcome and outcome prediction in the critically ill patients deserves further evaluation. PMID- 17116011 TI - Statin treatment, reduced T-cell content of atherosclerotic plaques and cancer. PMID- 17116013 TI - Gains and losses during anticoagulation treatment in patients with mechanical heart valves. PMID- 17116014 TI - Bleeding in patients with mechanical heart valves on warfarin: impact of target INR and of comorbidity. PMID- 17116015 TI - The blood-brain barrier and epilepsy. AB - During the past several years, there has been increasing interest in the role of the blood-brain barrier (BBB) in epilepsy. Advances in neuroradiology have enhanced our ability to image and study the human cerebrovasculature, and further developments in the research of metabolic deficiencies linked to seizure disorders (e.g., GLUT1 deficiency), neuroinflammation, and multiple drug resistance to antiepileptic drugs (AEDs) have amplified the significance of the BBB's relationship to epilepsy. Prior to 1986, BBB research in epilepsy focused on three main areas: ultrastructural studies, brain glucose availability and transport, and clinical uses of AEDs. However, contrast-based imaging techniques and medical procedures such as BBB disruption provided a framework that demonstrated that the BBB could be reversibly disrupted by pathologic or iatrogenic manipulations, with important implications in terms of CNS drug delivery to "multiple drug resistant" brain. This concept of BBB breakdown for therapeutic purposes has also unveiled a previously unrecognized role for BBB failure as a possible etiologic mechanism in epileptogenesis. Finally, a growing body of evidence has shown that inflammatory mechanisms may participate in the pathological changes observed in epileptic brain, with increasing awareness that blood-borne cells or signals may participate in epileptogenesis by virtue of a leaky BBB. In this article we will review the relationships between BBB function and epilepsy. In particular, we will illustrate consensus and divergence between clinical reality and animal studies. PMID- 17116016 TI - Paroxysmal motor disorders of sleep: the clinical spectrum and differentiation from epilepsy. AB - The diagnosis of paroxysmal events in sleep represents a significant challenge for the clinician, with the distinction of nocturnal epilepsy from nonepileptic sleep disorders often the primary concern. Diagnostic error or uncertainty is not uncommon in this situation, particularly with respect to nocturnal frontal lobe epilepsy (NFLE), which has a variable and often unusual presentation. Such errors can be minimized if the range of nonepileptic disorders with motor activity in sleep is fully appreciated. Here we review these disorders, before discussing the important clinical and electrographic features that allow their accurate differentiation from seizures. Particular emphasis is placed on the differentiation of nocturnal frontal lobe epilepsy from non-rapid eye movement (NREM) arousal disorders and other parasomnias. The value of recording episodes with video EEG polysomnography is discussed. PMID- 17116017 TI - Epidural pentobarbital delivery can prevent locally induced neocortical seizures in rats: the prospect of transmeningeal pharmacotherapy for intractable focal epilepsy. AB - PURPOSE: To determine whether epidural pentobarbital (PB) delivery can prevent and/or terminate neocortical seizures induced by locally administered acetylcholine (Ach) in freely moving rats. METHODS: Rats were implanted permanently with an epidural cup placed over the right parietal cortex with intact dura mater. Epidural screw-electrodes, secured to the cup, recorded local neocortical EEG activity. In the seizure-termination study, Ach was delivered into the epidural cup, and after the development of electrographic and behavioral seizures, the Ach solution was replaced with either PB or artificial cerebrospinal fluid (aCSF; control solution). In the seizure-prevention study, the epidural Ach delivery was preceded by a 10-min exposure of the delivery site to PB or aCSF. Raw EEG recordings, EEG power spectra, and behavioral events were analyzed. RESULTS: Ach-induced EEG seizures associated with convulsions, which were unaffected by epidural aCSF applications, were terminated by epidurally delivered PB within 2-2.5 min. Epidural deliveries of PB before Ach applications completely prevented the development of electrographic and behavioral seizures, whereas similar deliveries of aCSF exerted no influence on the seizure-generating potential of Ach. CONCLUSIONS: This study showed for the first time that epidural AED delivery can prevent, as well as terminate, locally induced neocortical seizures. The findings support the viability of transmeningeal pharmacotherapy for the treatment of intractable neocortical epilepsy. PMID- 17116018 TI - Effects of the novel antiepileptic drug lacosamide on the development of amygdala kindling in rats. AB - PURPOSE: The current treatment of epilepsy focuses exclusively on the prophylaxis or suppression of seizures and thus provides merely a symptomatic treatment, without clear influence on the course of the disease. There is a need for new drugs that act at different molecular targets than currently available antiepileptic drugs (AEDs) and for new therapies designed to block the process of epileptogenesis. In recent years, different research lines have examined the epileptogenic process in order to understand the different stages in this process, and with the hope that early recognition and intervention could prevent the development or progression of epilepsy. In animals, acquired epilepsy is studied most commonly with the kindling model and status epilepticus models. In the present study, we used the kindling model to evaluate whether the novel AED lacosamide affects kindling-induced epileptogenesis. This drug does not seem to act by any of the mechanisms of currently available AEDs, but the exact molecular mechanisms of action of lacosamide have not yet been clarified. METHODS: Groups of 9-10 rats were treated with either vehicle or different doses of lacosamide (3, 10, or 30 mg/kg/day) over 22-23 days during amygdala kindling. RESULTS: Daily administration of lacosamide during kindling acquisition produced a dose dependent effect on kindling development. While the drug was inactive at 3 mg/kg/day, significant retardation of kindling was observed at 10 mg/kg/day, by which the average number of stimulations to reach kindling criterion was increased by >90%. A significant inhibitory effect on kindling acquisition was also observed with 30 mg/kg/day, but this dose of lacosamide was associated with adverse effects. CONCLUSIONS: The present data demonstrate that lacosamide, in addition to exerting anticonvulsant activity, has the potential to retard kindling-induced epileptogenesis. Whether this indicates that lacosamide possesses antiepileptogenic or disease-modifying potential needs to be further evaluated, including studies in other models of acquired epilepsy. PMID- 17116019 TI - Anticonvulsant effect of Annona diversifolia Saff. and palmitone on penicillin induced convulsive activity. A behavioral and EEG study in rats. AB - PURPOSE: To evaluate hypnotic and anticonvulsant activities of Annona diversifolia Saff. and palmitone by using behavior and electroencephalographic (EEG) analysis in an experimental model of focal seizures in rats. METHODS: For hypnotic assessment, EEG analysis of polysomnographic slow-wave sleep (SWS) and rapid eye movement (REM) sleep for a 1 h period were performed after vehicle, A. diversifolia extract or palmitone, administration. For anticonvulsant effect, 60 minutes after treatments, EEG and behavior were analyzed during penicillin induced seizures. Latency to the onset of the first paroxystic spike, first seizure and frequency, as well as seizure severity using Racine's scale, were determined. RESULTS: Palmitone, but not A. diversifolia extract, produced a delay in the latency to the SWS phase. In addition, both palmitone and extract decreased SWS duration and accumulated REM sleep phase. With regard to the seizures, both the extract and palmitone increased the latency to the onset of spikes and seizures, but also decreased the duration of penicillin-induced seizures. This reduction in the EEG recordings was associated with an attenuation in the severity of behavioral seizures. CONCLUSIONS: A. diversifolia and palmitone did not produce a sedative-hypnotic effect although both of them were effective in reducing the severity of behavioral and EEG seizures induced by penicillin in rats, suggesting that the diminution in the paroxystic activity by A. diversifolia is likely produced by palmitone through GABAergic neurotransmission. This study justifies and reinforces the traditional use of this plant in epilepsy. PMID- 17116020 TI - In situ metabolism of levetiracetam in blood of patients with epilepsy. AB - PURPOSE: Although levetiracetam undergoes minimum metabolism, B-esterases have been identified in whole blood that are capable of metabolising levetiracetam. The present study was designed to ascertain any variability in levetiracetam blood concentrations that could be attributed to in situ metabolism and which could impact on the utility of such concentration measurements in guiding therapeutic management. METHODS: Blood samples were collected from 40 patients that were prescribed levetiracetam. Sera (Groups 1 and 2) or whole blood (Groups 3 and 4) were compared. Paraoxan, an inhibitor of B-esterase activity, was added to samples assigned to Groups 2 and 4. Samples within each group were assigned to Time 0 (frozen within 30 min of sample collection), Time 2 days and Time 7 days (samples kept at ambient temperature for 2 and 7 days). RESULTS: For serum samples, mean levetiracetam concentrations at Time 2 days and Time 7 days were indistinguishable from Time 0, regardless of whether B-esterase activity was inhibited on not. In contrast, for whole blood, in the absence of B-esterase inhibition, mean levetiracetam concentrations declined over time (11% and 29%; 2 and 7 days) compared to baseline values. In the presence of B-esterase inhibitor, mean levetiracetam concentrations at 2 days were indistinguishable from baseline values, although at 7 days values declined by 4%. CONCLUSIONS: If therapeutic monitoring of levetiracetam is to be undertaken, serum should be the matrix of choice and that whole blood should be separated as soon as possible after patient sampling so as to minimize in situ levetiracetam metabolism which could result in spuriously low concentrations and substantial intrapatient variability. PMID- 17116021 TI - Pharmacokinetics of the new antiepileptic and CNS drug RWJ-333369 following single and multiple dosing to humans. AB - PURPOSE: To characterize the pharmacokinetics of the new antiepileptic and CNS drug RWJ-333369 following single and multiple oral doses to healthy subjects, including the effect of food on bioavailability. METHOD: Two studies were conducted. The first study had a randomized, double-blind, placebo-controlled, sequential, ascending-dose crossover design. Subjects were divided into four dose groups (100, 250, 500, and 750 mg) of 10 to 11 subjects each. RWJ-333369 or placebo was administered for two 7-day periods, separated by a 14-day washout. In the second study RWJ-333369 (750 mg) was administered to 12 healthy subjects under fasted and fed conditions. Plasma and urine samples were analyzed for RWJ 333369 by liquid chromatography-mass spectroscopy. Safety was assessed throughout the studies. RESULTS: Mean (range) pharmacokinetic parameters in the above studies were: oral clearance (CL/F) 3.4-4.2 L/h, half-life (t(1/2)) 10.6-12.8 h, and renal clearance (CLr) 0.042-0.094 L/h, indicating that RWJ-333369 is eliminated primarily by metabolism. These parameters were not significantly different (p > 0.05) for the four dose groups and for single and multiple dosing. C(max) and AUC increased proportionally with dose and decreased with food by 11% and 5%, respectively. CONCLUSIONS: Following single and repetitive (q12h) doses of 100-750 mg, RWJ-333369 had linear pharmacokinetics; food did not alter pharmacokinetics to a clinically relevant extent. RWJ-333369 is extensively metabolized and has a low CL/F that equals < 5% of the liver blood flow. Thus, orally administered RWJ-333369 has no hepatic first-pass effect. The 12-h half life will enable bid dosing with an immediate-release oral formulation. PMID- 17116022 TI - Pharmacokinetic interaction study between the new antiepileptic and CNS drug RWJ 333369 and carbamazepine in healthy adults. AB - PURPOSE: To characterize the possible pharmacokinetic interaction between the new antiepileptic and CNS drug RWJ-333369 and carbamazepine (CBZ) following multiple dosing in healthy subjects. METHODS: In an 8-week, open-label, sequential design study, 24 healthy adults received multiple-dose RWJ-333369 alone (5 days 250 mg q12h; 5 days 500 mg q12h), then after a 4-day washout, multiple-dose CBZ alone (3 days 100 mg q12h; 3 days 200 mg q12h; 22 days 300 mg q12h), and then combination of CBZ (300 mg q12h), and RWJ-333369 (5 days 250 mg q12h; 5 days 500 mg q12h). RESULTS: At steady-state following multiple dosing, RWJ-333369 peak plasma concentration (C(max)) and area under the concentration-time-curve within the dosing interval (AUCss) increased in proportion to dose. The C(max) and AUCss of CBZ were similar when given alone or concomitantly with RWJ-333369. The 90% confidence intervals for the ratio of CBZ C(max) and AUCss with/without RWJ 333369 were: 94-104% and 95-104%, respectively (well within the equivalence range of 80-125%). When RWJ-333369 was administered with CBZ, its mean (SD) oral clearance increased from 3.2 L/h to 4.9 L/h and consequently its mean half-life was shortened from 10.4 (1.9) h to 7.4 (1.2) h, and mean AUCss and C(max) were reduced by 37% and 30%, respectively. CONCLUSIONS: There was no effect of multiple-dose RWJ-333369 on CBZ pharmacokinetics. CBZ induced RWJ-333369 clearance, resulting in shortened half-life and decreased exposure (AUCss) and C(max). Concomitant administration of RWJ-333369 with CBZ was generally safe and tolerated. PMID- 17116023 TI - Characterization of the anticonvulsant, behavioral and pharmacokinetic interaction profiles of stiripentol in combination with clonazepam, ethosuximide, phenobarbital, and valproate using isobolographic analysis. AB - PURPOSE: Isobolographic analysis was used to characterize the interactions between stiripentol (STP) and clonazepam (CZP), ethosuximide (ETS), phenobarbital (PB), and valproate (VPA) in suppressing pentylenetetrazole (PTZ)-induced clonic seizures in mice. METHODS: The anticonvulsant and acute adverse (neurotoxic) effects of STP in combination with the various conventional antiepileptic drugs (AEDs), at fixed ratios of 1:3, 1:1, and 3:1, were evaluated in the PTZ and chimney tests in mice using the isobolographic analysis. Additionally, protective indices (PI) and benefit indices (BI) were calculated to identify their pharmacological profiles so that a ranking in relation to advantageous combination could be established. Moreover, adverse-effect paradigms were determined by use of the step-through passive avoidance task (long-term memory), threshold for the first pain reaction, grip-strength test (neuromuscular tone), and the hot plate test (acute thermal pain). Brain AED concentrations were also measured so as to ascertain any pharmacokinetic contribution to the pharmacodynamic interactions. RESULTS: All AED combinations comprising of STP and CZP, ETS, PB, and VPA (at the fixed ratios of 1:3, 1:1 and 3:1) were additive in terms of clonic seizure suppression in the PTZ test. However, these interactions were complicated by changes in brain AED concentrations consequent to pharmacokinetic interactions. Thus STP significantly increased total brain ETS and PB concentrations, and decreased VPA concentrations, but was without effect on CZP concentrations. In contrast, PB significantly decreased and VPA increased total brain STP concentrations while CZP and ETS were without effect. Furthermore, while isobolographic analysis revealed that STP and CZP in combination, at the fixed ratios of 1:1 and 3:1, were supraadditive (synergistic; p < 0.05), the combinations of STP with CZP (1:3), ETS, PB, or VPA (at all fixed ratios of 1:3, 1:1, and 3:1) were barely additivity in terms of acute neurotoxic adverse effects in the chimney test. Additionally, none of the examined combinations of STP with conventional AEDs (CZP, ETS, PB, VPA--at their median effective doses from the PTZ-test) affected long-term memory, threshold for the first pain reaction, neuromuscular tone, and acute thermal pain. CONCLUSIONS: Based on BI values, the combination of STP with PB at the fixed ratio of 1:3 appears to be a particularly favourable combination. In contrast, STP and CZP or ETS (at the fixed ratios of 1:1 and 3:1) were unfavorable combinations. However, these conclusions are confounded by the fact that STP is associated with significant pharmacokinetic interactions. The remaining combinations of STP with PB (1:1 and 3:1), CZP (1:3), ETS (1:3), and VPA (at all fixed ratios of 1:3, 1:1, and 3:1) do not appear to be potential favorable AED combinations. PMID- 17116024 TI - The evaluation of thyroid functions, thyroid antibodies, and thyroid volumes in children with epilepsy during short-term administration of oxcarbazepine and valproate. AB - PURPOSE: The aim of this study was to evaluate the effects of short-term oxcarbazepine (OXC) and valproate (VPA) monotherapy on thyroid functions in children. METHODS: Fifty-five newly diagnosed epileptic children with normal thyroid functions (confirmed with the thyrotropin releasing hormone stimulation test) participated in this study. VPA treatment was started in 30 patients and OXC in 25 patients. Serum thyroxine (T(4)), free thyroxine (fT(4)), triiodothyronine (T(3)), free triiodothyronine (fT(3)), reverse T3 (rT(3)), thyroid peroxidase antibodies (TPO-ab), and urine iodine levels were evaluated at baseline and at the third and sixth months of therapy. RESULTS: In the OXC group, serum T(4), fT(4), T(3), fT(3), and rT(3) levels were found to be decreased at the third and sixth months, the differences were significant compared to the baseline values except for fT(3) levels at the third month and fT(4) and rT(3) levels at the sixth month (p < 0.05). At the sixth month, serum T(4) level dropped below the normal reference value in 8 (32%), fT(4) in 5 (20%), T(3) in 4 (16%), and fT(3) in 3 (12%) patients. In the VPA group, mean T(4), fT(4), T(3), fT(3), and rT(3) levels at 3 and 6 months remained similar compared to the baseline values (p > 0.05). Mean serum thyroid stimulating hormone levels increased significantly at the sixth month compared to the baseline values in the VPA group (p < 0.05) while it remained unchanged in the OXC group (p > 0.05). There was no effect of either drug on urinary iodine excretion and serum TPO-ab levels remained in normal ranges throughout the study. CONCLUSIONS: In this prospective study, it is documented that children under short-term OXC or VPA therapy showed altered thyroid functions similar to the changes observed after long-term treatment. Although, the clinical significance of these results need to be evaluated with future studies, this observation of altered thyroid functions points out that thyroid functions may need to be monitored closely in children receiving antiepileptic treatment, even in the short-time interval. PMID- 17116025 TI - A long-term follow-up of zonisamide monotherapy. AB - OBJECTIVES: Several studies have reported the safety and efficacy of zonisamide monotherapy, but studies on its long-term outcomes are limited. This chart review was conducted to evaluate the long-term outcomes of zonisamide monotherapy. METHODS: The charts were reviewed for 77 patients treated with zonisamide as monotherapy for 6-180 months between May 1985 and December 2003. Outcomes were analyzed by the following subcategories: patients with newly diagnosed epilepsy or with antiepileptic drug-resistant epilepsy, the type of epilepsy, patient age, and treatment period. RESULTS: Of a total of 77 patients, 49 patients (64%) attained 50% or more reduction of seizure frequency and of those patients 38 (49%) attained 75% or more reduction, with 18 patients (24%) becoming seizure free from 6 to 180 (median 80.6 +/- 43.6) months of follow-up. Thirty-eight patients (49%) continued zonisamide monotherapy as of December 2003. Proportions of patients having 75% or more reduction in seizure frequency in subcategories were as follows; 56% in patients with newly diagnosed epilepsy and 48% in patients whose treatment was switched to zonisamide monotherapy owing to lack of efficacy of or adverse reaction to previous antiepileptic drugs; 60% in patients with localization-related epilepsies and 38% in patients with generalized epilepsies; and 49% in pediatric patients and 50% in adult patients. CONCLUSION: Long-term zonisamide monotherapy was efficacious in a wide range of patients with epilepsy. Zonisamide did not seem to exhibit a reduction in efficacy during long term use of up to 180 months. PMID- 17116026 TI - Assessment of the long-term effects of epilepsy surgery with three different reference groups. AB - PURPOSE: It is difficult to validly assess the long-term effect of epilepsy surgery. Here, this is attempted by comparing the outcome of surgically treated pharmacoresistant epilepsy patients to three different nonoperated comparison groups regarding seizure control, antiepilepsy drug (AED) usage, and health related quality of life (QOL). METHODS: One hundred thirty-one operated patients (group 1, mean follow-up since presurgical assessment 6.9 years), 105 patients awaiting presurgical assessment (group 2, mean follow-up after assignment to waiting list 0.8 years), 99 patients considered to be presurgical candidates who chose to withdraw from waiting for presurgical assessment (group 3, mean follow up after assignment to waiting list 5.5 years), and 49 patients who were not deemed to be eligible for surgery after comprehensive assessment (group 4, mean follow-up since presurgical assessment 6.5 years) were studied. The patients completed a questionnaire on seizures, AED usage, and QOL (ESI-55). RESULTS: The surgical patients had a better outcome than all three comparison groups regarding seizure frequency, seizure freedom rate, and number of AEDs used. They scored higher than groups 2, 3, and 4 on 7/11, 6/11, and 3/11 ESI-55 domains, respectively. CONCLUSIONS: The superior long-term outcome of the operated patients was most marked if compared to the patients awaiting surgery. This is compatible with the assumption that patients present for presurgical candidacy selection and assessment at a "nadir" of their disease course. After several years, a regression to the mean occurs which reduces (but does not abolish) the differences between nonoperated and operated patients. PMID- 17116027 TI - Predictors of neuropsychological impairment in seizure-free epilepsy patients. AB - Persons with epilepsy are at increased risk of cognitive deficits as a result of various factors like etiology, structural brain lesions, seizure frequency, seizure type, age at onset of epilepsy, hereditary factors, psychosocial factors, and possible adverse effects of antiepileptic drugs (AEDs). Despite the fact that the majority of epilepsy patients are seizure-free, previous studies on the relationship between epilepsy-related variables and cognitive function have mainly been conducted on patients with persisting seizures. In this study 158 adults with epilepsy on AED monotherapy and without epileptic seizures for at least 2 years were investigated with a neuropsychological test battery in addition to a neurological examination, MRI and EEG. The major findings were that the group had education and employment status similar to the population mean and neuropsychological function in the normal range. In the patient group without idiopathic generalized epilepsy known cerebral etiology was found to be a highly significant predictor of neuropsychological deficit. For patients with idiopathic generalized epilepsy, early seizure debut at < or =18 years was a powerful predictor of neuropsychological impairment. PMID- 17116028 TI - Cognitive impairment is not equal in patients with epileptic and psychogenic nonepileptic seizures. AB - PURPOSE: Patients with psychogenic nonepileptic seizures (PNES) and those with epileptic seizures (ES) purportedly have roughly equal neurocognitive deficits. However, recent findings suggest that patients with somatoform disorders exhibit more variable effort on neurocognitive testing than do controls. We reexamined neurocognitive function in patients with ESs and PNES by using symptom validity testing to control for variability in effort. METHODS: Patients referred for video-EEG monitoring were administered the Word Memory Test (WMT), a measure of symptom validity, as part of neuropsychological evaluation. Patients classified with ictal video-EEG recordings as having ES (n = 41) or PNES (n = 43) were compared on neurocognitive and WMT performance and demographic, psychiatric, and medical variables. RESULTS: Striking rates of WMT failure were observed in the PNES (51.2%) group, but not in the ES (8.1%) group (p = <0.001) after controlling for false-positive errors. Although the PNES and ES groups reported equivalent neurologic histories, the PNES group exhibited less objective evidence of impairment as measured by valid neuropsychological testing, MRI of the brain, and video-EEG monitoring. CONCLUSIONS: Many patients with PNES do not put forth maximal effort during neuropsychological assessment. When patients with PNES put forth valid effort, they demonstrate less objective evidence of neuropathologic injury or disease than do patients with ES. The cognitive impairment reported by this group appears to be more a function of motivational (although not necessarily intentional) factors than of verifiable neuropathology. PMID- 17116029 TI - Predicting memory decline following epilepsy surgery: a multivariate approach. AB - BACKGROUND: While some patients experience a decline in memory function following an anterior temporal lobe resection, there is considerable individual variation in the extent, nature, and direction of postoperative memory change. Patients with surgically remediable temporal lobe epilepsy differ in etiology, the extent and type of underlying pathology, and on demographic and epilepsy-related variables, all of which may have an impact on their pre- and postoperative neuropsychological functioning. This study examined the relationship between these variables and postoperative memory decline. METHODS: Logistic regression was used to examine the effects of age, laterality of surgery, age of onset of epilepsy, underlying pathology and preoperative level of memory function on postoperative verbal learning in 288 patients who had undergone an anterior temporal lobe resection. One hundred twenty-five patients underwent a right temporal lobe resection (RTL), 163 patients underwent a left temporal lobe resection (LTL). RESULTS: In the group as a whole, 25% of the patients demonstrated a significant postoperative deterioration in verbal learning. Postoperative deterioration in verbal learning was significantly associated with higher levels of preoperative function in both the RTL and LTL groups. Older age at the time of the operation and a lower verbal IQ were additional significant predictors for the RTL group. The presence of cortical dysgenesis was a significant predictor of postoperative decline in the LTL group. The logistic regression models accurately identified 3/4 of those who experienced a postoperative decline in memory, using a cutoff of 0.25 or above to identify high risk. CONCLUSIONS: Our analyses suggest that the majority of patients with a high risk of significant postoperative memory decline can be reliably identified preoperatively. These models are valuable tools helping patients make an informed decision regarding surgery. PMID- 17116030 TI - Evaluating the contributions of state-of-the-art assessment techniques to predicting memory outcome after unilateral anterior temporal lobectomy. AB - PURPOSE: Although anterior temporal lobectomy (ATL) is an effective treatment for many patients with medically refractory temporal lobe epilepsy (TLE), one risk associated with this procedure is postsurgical decline in memory. A substantial number of past studies examined factors that predict memory decline after surgery, but few have investigated multiple predictors simultaneously or considered measures that are currently in use. METHODS: This study compared the relative contributions made by presurgical neuropsychological test scores, MRI based hippocampal volumetric analysis, and Wada test results to predicting memory outcome after ATL in a group of 87 patients. RESULTS: Logistic regression analyses indicated that noninvasive procedures (neuropsychological testing and MRI) made significant contributions to improving the prediction of memory outcome in this sample. The results from the Wada procedure did not significantly improve prediction once these other factors were considered. The only exception was in predicting memory for visual information after a delay, in which Wada results improved prediction accuracy from 78% to 81%. CONCLUSIONS: Current neuropsychological tests and MRI volumetric measures predict changes in verbal and visual memory after ATL. The relatively small change in correct classification rates when Wada memory scores are considered calls into question the benefits of using Wada test results to predict memory outcome when the results of noninvasive procedures are available. PMID- 17116031 TI - Face memory in MRI-positive and MRI-negative temporal lobe epilepsy. AB - PURPOSE: Effects of MRI-positive (MRI(+)) as compared to MRI-negative (MRI(-)) temporal lobe epilepsy (TLE) on face memory are not yet known. METHODS: We studied 24 MRI(-) (11 right/13 left) and 20 MRI(+) (13 right/7 left) TLE patients, 12 generalized epilepsy patients, and 12 healthy subjects undergoing diagnostic workup with 24-72-h Video-EEG-monitoring. Twenty faces were shown, and had to be recognized from 40 faces immediately and after a 24-h delay. RESULTS: MRI(+) and MRI(-) right TLE (RTLE) patients showed deficits in face recognition compared to controls or generalized epilepsy, consistent with right temporal lobe dominance for face recognition. MRI(+) RTLE patients had deficits in both immediate and delayed recognition, while MRI(-) RTLE patients showed delayed recognition deficits only. The RTLE groups showed comparable delayed recognition deficits. Separate analyses in which the MRI(+) group included patients with hippocampal sclerosis only, did not alter results. Furthermore, MRI(-) RTLE had a worse delayed recognition than MRI(-) left TLE (LTLE). On the other hand, MRI(+) RTLE did not differ from MRI(+) LTLE in delayed recognition. Combining MRI(-) and MRI(+) TLE groups, we found differences between RTLE and LTLE in delayed, but not immediate face recognition. CONCLUSIONS: Our results suggest that a delayed recognition condition might be superior to immediate recognition tests in detecting face memory deficits in MRI(-) RTLE patients. This might explain why former studies in preoperative patients did not observe an immediate face recognition dominance of the right temporal lobe when combining MRI(-) and MRI(+) TLE patients. Our data further point to an important role of the right mesial temporal region in face recognition in TLE. PMID- 17116032 TI - Prevalence of self-reported epilepsy or seizure disorder and its associations with self-reported depression and anxiety: results from the 2004 HealthStyles Survey. AB - PURPOSE: To examine the prevalence of self-reported epilepsy or seizure disorder and its association with self-reported recent depression and anxiety in a large sample of the U.S. adult population. METHODS: We analyzed data from adults aged 18 years or older (n = 4,345) who participated in the 2004 HealthStyles Survey, a large mail panel survey designed to be representative of the U.S. population. RESULTS: Among U.S. adults aged 18 years or older, we estimated that 2.9% have been told by a doctor that they had epilepsy or seizure disorder, and an estimated 1.6% and 0.9% had active and inactive epilepsy, respectively. After controlling for demographic characteristics, we estimated that adults with self reported epilepsy were twice as likely to self-report depression or anxiety in the previous year as were adults without epilepsy, and adults with active epilepsy were 3 times as likely to self-report depression and twice as likely to have anxiety in the previous year as were adults without epilepsy. CONCLUSIONS: Our findings highlight the burden of self-reported depression and anxiety among adults with self-reported epilepsy or seizure disorder, and suggest that healthcare providers should attempt to determine whether adult patients with epilepsy have any psychiatric comorbidity potentially to improve health outcomes. Questions about epilepsy and related factors should be routinely included on population-based surveys so that we can better understand the epilepsy distribution in the U.S. population and identify the unmet health and psychosocial needs of people with epilepsy. PMID- 17116033 TI - Memory, emotional and vocational impairments before and after anterior temporal lobectomy for complex partial seizures. AB - PURPOSE: To assess the pre- and postsurgical frequency of memory, emotional, and vocational impairments in patients who underwent anterior temporal lobectomy (ATL), and to assess the relationship between emotional disturbance and memory abilities after ATL. METHODS: Retrospective analysis of data was performed on 90 patients with medically intractable complex partial seizures who underwent ATL between 1981 and 2003. Patients were evaluated an average of 5 months before surgery and 11.3 months after surgery. RESULTS: A moderate to high frequency of memory impairment (44.4%; verbal or nonverbal), emotional disturbance (38.9%) and unemployment (27.8%) existed in the same individuals both before and after surgery. There were small to moderate rates of new onset memory (18.9%), emotional (11.1%), and vocational (7.8%) difficulties after surgery often regardless of seizure control outcome. Patients who underwent left-ATL and had emotional disturbance after surgery had the lowest verbal memory test scores. CONCLUSIONS: Results highlight the importance of taking into account emotional status when assessing memory abilities after ATL. Results replicate the finding of moderate to high frequencies of memory impairment, emotional disturbance, and unemployment both before and after ATL. Results provide support for the rationale that cognitive, psychiatric and vocational interventions are indicated to mitigate the problems that exist before and persist after ATL. PMID- 17116034 TI - Patient awareness of complex partial seizures. AB - PURPOSE: To assess self-awareness of complex partial seizures (CPSs) in unselected epilepsy patients through a thorough interview. METHODS: The study comprised 134 patients at our epilepsy clinic, whose CPSs had been documented by the patient's family members. We investigated the proportion and characteristics of patients unaware of their CPSs compared to those who were, and we monitored the evolution of unawareness of CPSs during the follow-up. RESULTS: Thirty-one (23%) patients were assigned to the unawareness group (complete, 23; incomplete, 8) and 103 (67%) patients to the awareness group. Patients in the unawareness group were older and had a later age of onset than patients in the awareness group. Interictal epileptiform discharges (IEDs) localized primarily to the temporal region and were more frequently detected in the unawareness group (94%) than the awareness group (55%). Bilateral independent IEDs were found more frequently in the unawareness group than in the awareness group (48% vs. 13%). The bilateral presence of lesions was also more frequent in the unawareness group than the awareness group (16.1% vs. 4.9%). Six (26%) of 23 patients with complete unawareness of their CPSs had experienced awareness of CPSs during the follow-up. Two of these patients even experienced the emergence of de novo aura. CONCLUSIONS: Our results indicate that a significant number of epilepsy patients are not aware of their CPSs. Unawareness of CPSs may be related to bitemporal dysfunction and a rapid and complete loss of consciousness caused by rapid spread of ictal discharges to the contralateral hemisphere in association with bilateral independent IEDs and bilateral presence of lesions. PMID- 17116035 TI - Executive dysfunction is a significant predictor of poor quality of life in children with epilepsy. AB - PURPOSE: Based on prior research indicating poor health-related quality of life (HRQOL) in children with attention-deficit/hyperactivity disorder, we investigated (1) whether executive functioning deficits were related to poor HRQOL in children with epilepsy, (2) how important these variables were in comparison to known predictors of HRQOL such as neurological factors, and (3) the extent to which clinical-level impairments in executive dysfunction predispose children to low HRQOL. METHOD: Data included scores on the Behavior Rating Inventory of Executive Function (BRIEF) and HRQOL scales (The Impact of Childhood Illness Scale [ICI] and Hague Restrictions in Epilepsy Scale [HARCES]) for 121 children (mean age = 11.9, SD = 3.6) from a tertiary center serving children with severe epilepsy. RESULTS: Correlations between the BRIEF and ICI total and subscore domains (child, parent, family, and treatment) were generally significant and moderate (e.g., r > or = 0.30, p < or = 0.001). BRIEF Global Executive Composite, number of antiepileptic drugs (AEDs), number of prior AEDs, and adaptive level all emerged as significant and unique predictors of HRQOL (R(2)= 0.36, adj. R(2)= 0.33, p < 0.0001). A clinically elevated BRIEF was associated with a twofold risk of low HRQOL (odds ratio = 2.21, p = 0.03). CONCLUSIONS: Executive dysfunction appears to exert a broad adverse influence on HRQOL in children with epilepsy, with clinical-level impairments in executive dysfunction contributing to a twofold increase in the likelihood of poor HRQOL. The constellation of executive dysfunction, low adaptive level, high medication load, and a history of several failed AEDs are risk factors for poor HRQOL in children with epilepsy. PMID- 17116036 TI - How to replace lamotrigine with valproate. AB - If lamotrigine (LTG) has to be replaced with valproate (VPA), this exchange may be complicated by adverse events that result from the complex interaction of both drugs. We report on two cases in which such problems occurred in spite of a cautious switch considering the VPA induced LTG serum increase. The satisfying outcome after a sudden and complete withdrawal of LTG in both cases encouraged us to perform the switch from LTG to VPA systematically by discontinuing LTG abruptly and building up the VPA maintenance dosage very rapidly in the following five consecutive patients who required this exchange. We recommend our abrupt dosage change-over strategy as an easy, safe and cost-effective option. PMID- 17116037 TI - Differential effects of antiepileptic drugs on neuroactive steroids in men with epilepsy. AB - PURPOSE: To compare serum levels of neuroactive steroids among men with epilepsy who take various antiepileptic drugs, untreated men with epilepsy and normal controls (NC). METHODS: Subjects were 85 men with localization-related epilepsy [unmedicated >6 months (No Rx)-10, carbamazepine (CBZ)-25, phenytoin (PHT)-25, lamotrigine (LTG)-25] and 25 NC. Sexual function scores (S-Score), hormone levels [dehydroepiandrosterone sulfate (DHEAS), bioactive (BA) testosterone (T), estradiol (BAE), and androstanediol (BAL)] and the ratios of inhibitory to excitatory neuroactive metabolites of T, i.e., BAL/BAE, were compared among groups. RESULTS: S-scores, DHEAS, and bioactive testosterone (BAT) were significantly (p < 0.05) lower and BAL and BAL/BAE were significantly higher among CBZ and PHT groups than among NC and LTG groups. LTG did not differ from NC in any of these measures. BAT correlated significantly with BAL/BAE for PHT (r = 0.44, p = 0.02) and CBZ (r = 0.42, p = 0.03) but not for NC (r = 0.03, p = NS) and LTG (r = 0.06, p = NS) groups. CONCLUSIONS: In comparison to LTG, enzyme inducing AEDs (CBZ, PHT) are associated with a more favorable neuroactive steroid balance (lower DHEAS and higher BAL/BAE) for seizure management, but at the expense of reduced serum bioavailable testosterone levels and sexual function. PMID- 17116038 TI - Prolonged focal negative motor seizures: a video-EEG study. AB - PURPOSE: Focal negative motor (akinetic) seizures are rare ictal events that are diagnostically challenging because they are difficult to differentiate from postictal Todd paresis, transient ischemic attacks, migraine events, and psychogenic episodes. METHODS: We describe a 45-year-old man in whom, after surgical drainage of a right frontoparietal subdural hematoma, prolonged episodes developed with flaccid paralysis of the left arm. RESULTS: A video-EEG recording demonstrated a close relation between the focal motor impairment and a clear-cut epileptic ictal discharge involving the right perirolandic cortical areas. CONCLUSIONS: On the basis of the electroclinical data, we hypothesize the involvement of the primary motor area in the genesis of the epileptic discharge triggering focal negative seizures. PMID- 17116039 TI - Topographic movie of ictal high-frequency oscillations on the brain surface using subdural EEG in neocortical epilepsy. AB - PURPOSE: To understand the rapid dynamic changes of ictal intracranial high frequency oscillations (HFOs) in neocortical epilepsy. METHODS: We integrated multiple band frequency analysis and brain-surface topographic maps of HFOs from ictal subdural EEG (SDEEG) recordings. We used SDEEG to record partial seizures consisting of right-arm jerks with secondary generalization in a 17-year-old right-handed girl. We selected 20-s EEG sections that included preclinical seizure recordings. We averaged the HFO power between 60 and 120 Hz for 25 selected electrodes, made topographic maps from these averaged powers, and superimposed the maps on the brain-surface image. We filmed consecutive HFO maps at a 10-ms frame rate. RESULTS: Before clinical seizure onset, high-power HFOs emerged at the superior portion of the left precentral gyrus, then appeared in the middle of the left postcentral gyrus, and subsequently reverberated between both regions as well as the posterior portion of the left postcentral gyrus. Right-arm extension and facial grimacing started as the HFO power decreased. As generalized tonic-clonic seizures evolved, HFO power increased but remained within the central region. CONCLUSIONS: Topographic movies of intracranial HFOs on the brain surface allow visualization of the dynamic ictal changes in neocortical epilepsy. PMID- 17116040 TI - Mesial temporal inhibition in a patient with deep brain stimulation of the anterior thalamus for epilepsy. AB - We investigated the electrophysiological effects of high-frequency anterior thalamic deep brain stimulation using intracerebral mesial and lateral temporal depth electrodes in a patient with intractable focal epilepsy. Monopolar and bipolar stimulation delivered to the thalamic anterior nucleus using the programmable ITREL II stimulation device led to a significant decrease of cross power spectral density and a nonsignificant decrease of coherence in ipsilateral hippocampal structures. No such effect was found in lateral temporal or contralateral sites. The hippocampal inhibition was clearly related to the voltage (> or =7 V) and frequency (> or =70 Hz) of the thalamic stimulus and occurred with a delay of approximately 60 s after stimulus onset. PMID- 17116041 TI - Neuromagnetic source localization of epileptiform activity in patients with graphogenic epilepsy. AB - PURPOSE: To clarify the source localization of epileptiform activity by using magnetoencephalography (MEG) in patients with graphogenic epilepsy. METHODS: MEG and simultaneous EEG were recorded with a 204-channel whole-head MEG system in two patients with graphogenic epilepsy. During the MEG recordings, the patients performed a set of tasks comprising mental arithmetic calculation, speaking, moving the right arm in a manner resembling writing, writing, and thinking of writing. Equivalent current dipoles (ECD) were calculated for epileptiform discharges on MEG by using a single-dipole model. The ECD were superimposed on the magnetic resonance images of the patients. RESULTS: The task of writing provoked seizures, in which both patients jerked the right arms. Thinking of writing also induced these seizures. In both patients, EEG associated with the seizures showed bursts of spike-and-slow-wave complexes predominantly in the centroparietal region. MEG also showed epileptiform discharges corresponding to the EEG bursts. ECDs obtained from the discharges were clustered in the left centroparietal area. CONCLUSIONS: Thinking of writing was a trigger for the seizures, as well as the task of writing. The source of the epileptiform discharge associated with the seizures was localized in the unilateral centroparietal area. The findings suggest that the centroparietal region plays an important role in the pathophysiology underlying these two graphogenic epilepsy cases. PMID- 17116042 TI - Neural network underlying ictal humming demonstrated by very early SPECT: a case report. AB - We report the case of a 49-year-old right-handed woman with brief partial seizures in which the clinical semiology was marked by an early humming automatism. MRI fusion of the registered ictal and interictal single-photon emission computed tomography (SPECT) substraction exhibited a left neural network involving lateral temporal, inferior frontal, and inferior parietal cortices. PMID- 17116043 TI - Toothbrush-thinking seizures. AB - Seizures associated with toothbrushing have been reported in patients with precentral or postcentral partial epilepsy. Seizures precipitated by thinking have been described in patients with idiopathic generalized epilepsy. We report a patient with intractable partial epilepsy in which seizures were induced both by toothbrushing, and by seeing or thinking about toothbrush and toothpaste. Video EEG analysis revealed a left temporal lobe origin for these reflex seizures. We discuss how complex multimodal stimuli may trigger these reflex seizures. PMID- 17116049 TI - Obstetric-induced incontinence: a black hole of preventable morbidity. AB - There is a detailed literature comprising clinical and anorectal physiological studies linking faecal incontinence to vaginal delivery. Specific risk factors are high infant birthweight, forceps delivery and prolonged second stage of labour. The onset of symptoms may be delayed for many years. Faecal incontinence occurs in more than 10% of adult females and urinary incontinence in about a third of multiparous women. This places a very large economic burden on the Australian health system. A conservative estimate for overall management of incontinence would be in excess of $A700 million but the actual amount is unknown. Preventative measures for avoiding pelvic floor injuries need to be established, and safe obstetric practice needs to be redefined in the light of current knowledge about incontinence. Outcome measures for safe birthing should not only include infant and maternal mortality and infant morbidity, but should also include the long-term effects of vaginal delivery on the pelvic floor, particularly urinary and faecal incontinence. Several state reports and one federal senate report on safe birthing have been lacking in this area. The safety of birthing centres and home birthing needs to be examined to provide birthing mothers with complete and appropriate information about safety in order that they may consider their options. Appropriate Caesarean section rates for optimal birthing safety are unknown and need to be re-examined. Calls for overall reduction in Caesarean section rates in Australia are inappropriate and cannot be justified until the effects of pelvic floor injury are added to the overall assessment. PMID- 17116050 TI - The Tissue Fixation System provides a new structural method for cystocoele repair: a preliminary report. AB - BACKGROUND: A new reconstruction principle that uses tensioned tapes instead of large mesh is described for cystocoele repair. AIM: To apply this method to patients with central, paravaginal and cervical ring defects. METHODS: Ninety patients, mean age 63 years (29-83) and mean weight 73 kg (52-117 kg), underwent cystocoele repair using the Tissue Fixation System (TFS). Tapes were applied as a retro-obturator U-sling (n=29), transversely between both arcus tendineus fascia pelvis (ATFP) ligaments (n=45), along the path of the cardinal ligament in patients with cervical ring defects (n=12), and longitudinally along the ATFP ligament (n=4). RESULTS: At mean eight months review (three to 15 months), two failures were reported. There was one haematoma that drained spontaneously at seven days, and there were no erosions. Mean hospital stay was one a half days for the Australian group (one to seven days) and five days (four to eight days) for the European group. After using single U-sling, one patient required intermittent catheterisation for seven days before she could pass urine freely. In one patient the bladder was perforated during dissection laterally towards the ATFP; the perforation was successfully repaired. CONCLUSIONS: The tensioned tape operation is simple and accurate, and appears to work well in the short term. Longer-term studies are required. PMID- 17116051 TI - Mifepristone- and misoprostol-induced mid-trimester termination of pregnancy: a review of 272 cases. AB - BACKGROUND: Mifepristone became available in New Zealand in 2001, and was first used for second trimester terminations at the Level J Unit, Wellington Hospital. The protocol is based on that published by Ashok et al. in Aberdeen. AIMS: To describe the use of mifepristone prior to misoprostol induction of labour for mid trimester termination and to compare outcomes with the published data. METHODS: A retrospective audit of prospectively collected notes for 272 women presenting for mid-trimester termination of pregnancy in a hospital termination clinic. Data collection included age, ethnicity, previous pregnancies, gestational age, induction-to-abortion interval, analgesia, and complications. Data were entered into an Access database and imported into Excel and Epi Info for the computation of descriptive statistics. RESULTS: Data on completed abortion were available for 271 women (one chose not to continue the abortion following mifepristone). The median time to abortion was 6 h, and mean number of doses of misoprostol was three. The proportion of women who aborted within 24 h was 95.9%. Immediate surgical evacuation of retained placenta was required in 22 women (8.1%). Heavy bleeding occurred in 22 women (8.1%), and seven required a transfusion (2.6%). The proportion of women who required parenteral narcotics was 78.2%. CONCLUSIONS: Outcomes for women in the present review were comparable with those for previous publications using the same regimen, with the exception of a higher transfusion rate. Our experience supports the finding that the use of mifepristone as pretreatment to misoprostol results in a shorter induction-to-delivery interval than the use of misoprostol alone as has been reported by other groups. PMID- 17116052 TI - Serum advanced oxidation protein products, myeloperoxidase and ascorbic acid in pre-eclampsia and eclampsia. AB - BACKGROUND: Activation products from neutrophils and the complement system might cause endothelial dysfunction, which is central to the aetiology of pre eclampsia. This study aimed to investigate the activity of myeloperoxidase (MPO), and its association with advanced oxidation protein products (AOPP), in women with pre-eclampsia and eclampsia. MATERIALS AND METHOD: Twenty-one pregnant women with pre-eclampsia, 11 pregnant women with eclampsia and 19 healthy pregnant women were studied. Serum levels of malondialdehyde (MDA), AOPP, ascorbic acid (AA) and activities of MPO and catalase (CAT) were measured using a colorimetric method. RESULTS: The MDA level was significantly higher in the pre-eclampsia (3.15+/-0.28 nmol/mL) and eclampsia (4.01+/-0.66 nmol/mL) groups than in controls (1.85+/-0.18 nmol/mL); the difference between MDA levels in the pre-eclampsia and eclampsia groups was not statistically significant. MPO activity was significantly higher in the eclampsia (347.59+/-88.06 U/L) group than in the pre eclampsia (196.17+/-30.8) and control (93.22+/-9.52) groups, and there was also no significant difference in these levels between the pre-eclampsia and control groups. CAT activity was significantly higher in the pre-eclampsia (166.35+/ 31.75 U/L) and eclampsia (166.98+/-40.31 U/L) groups than in controls (81.28+/ 7.41 U/L), and AA level was significantly higher in the pre-eclampsia (0.54+/ 0.15 mg/dL) group than in controls (0.18+/-0.01 mg/dL); the differences in AA and CAT activity between the pre-eclampsia and eclampsia groups were not statistically significant. AOPP levels did not change significantly among the control, pre-eclampsia and eclampsia groups (106.88+/-5.62, 98.89+/-6.47, 111.89+/-6.8 micromol/L, respectively). CONCLUSIONS: We suggest that increased oxidative stress might contribute to the pathophysiological mechanisms of pre eclampsia and eclampsia, and that AA and CAT might have a protective role via free radical-scavenging properties. However, further study is needed. PMID- 17116053 TI - Mapping uptake of prenatal diagnosis for Down syndrome and other chromosome abnormalities across Victoria, Australia. AB - OBJECTIVE: To investigate geodemographic characteristics of women having prenatal diagnosis for Down syndrome and other fetal chromosome abnormalities in Victoria, Australia. METHODS: A descriptive analysis of population-based data on all confinements, amniocenteses and chorionic villus sampling for 1998 and 2002 was undertaken. Multivariate logistic regression analysis of 2002 data investigated geographical differences, including selected maternal features and a socioeconomic status measure that may be associated with uptake of diagnostic testing. RESULTS: After adjusting for age, parity of three or more children was associated with a significantly decreased likelihood of testing (P<0.001). Women from Africa were 61% less likely to have testing than women born in Australia (P<0.01). While overall uptake of testing was lower than average in rural regions, only the Loddon Mallee had a significantly decreased likelihood of testing (OR 0.71 (0.53, 0.95), P=0.02). A socioeconomic index of place of residence showed no association with uptake. Metropolitan and rural women giving birth in private hospitals were 31 and 60% more likely to have a test than women giving birth in public hospitals (P<0.01). CONCLUSION: Geographical differences influence uptake of prenatal diagnosis, probably related to access to services. However, other maternal demographic factors also play a role in uptake. To ensure equity in access and autonomy in women's reproductive choices, reasons for exceptionally low uptake in certain localities or within certain subgroups of pregnant women should be investigated further, followed by appropriate changes in service provision. PMID- 17116054 TI - Random urine protein to creatinine ratio as a diagnostic method of significant proteinuria in pre-eclampsia. AB - BACKGROUND: Because of the importance of pre-eclampsia and proteinuria in pregnancy, a faster and simpler diagnostic method is needed. AIM: To compare random urine protein to creatinine ratio (p:c ratio) with 24-h urine protein excretion rate in pregnant women with a suspicion of pre-eclampsia. METHODS: The study was conducted on 100 pregnant women with gestational ages of >or=20 weeks; 50 patients were suspected of having pre-eclampsia and 50 were healthy pregnant women. A random urine sample for p:c ratio determination and a 24-h urine sample for protein measurement were obtained. RESULTS: All women suspected of having pre eclampsia had significant proteinuria. The single-voided p:c ratio demonstrated a sensitivity of 94% with a specificity of 96% at the cut-off>or=0.2 mg/mg. There was strong correlation between the two methods in this group (r=0.70, P<0.001, R2=49%). Using the same cut-off in the pregnant women who were not thought to have pre-eclampsia, the sensitivity of the test (p:c ratio) was 29% and the specificity was 87%. Pearson's correlation coefficient was 26 (P<0.06). Negative predictive value and positive predictive value were 34 and 83%, respectively. CONCLUSION: There is a strong correlation between 24-h urine protein excretion and single-voided urine p:c ratio in women suspected of having pre-eclampsia. A single-voided p:c ratio of >or=0.2 mg/mg is highly predictive for significant proteinuria. However, this test was not found to be a reasonable alternative to 24-h urine collection; and it must be followed by 24-h urine collection in a clinically suspect patient with a p:c ratio of <0.2 mg/mg. PMID- 17116055 TI - Ultrasound and clinical predictors for Caesarean delivery after labour induction at term. AB - OBJECTIVE: To assess the relationship of ultrasound assessment for amniotic fluid, fetal weight, cervical length, cervical funneling and clinical factors on the risk of Caesarean delivery after labour induction at term. METHODS: On hundred and fifty-two women scheduled for labour induction at term agreed to participate. Sonography was performed to obtain fetal biometry, amniotic fluid index and cervical length and to detect funneling at the internal cervical os. The sonographic findings were concealed. Study women received standard care during labour induction. RESULTS: On univariate analysis using Fisher's exact test, parity, cervical length and Bishop score were associated with Caesarean delivery. Following multivariable logistic regression analysis, only nulliparity (adjusted odds ratio (AOR) 5.2 (95% CI 2.2-12.2): P<0.001) and transvaginal ultrasound-determined cervical length of more than 20 mm (AOR 2.8 (95% CI 1.0 7.4): P=0.04) were independent predictors of Caesarean delivery in labour induction. Maternal age, maternal height, gestational age, indication for labour induction, amniotic fluid index, cervical funneling and ultrasound-estimated fetal weight did not predict Caesarean delivery. CONCLUSION: In women who had undergone labour induction at term with a singleton fetus, nulliparity and cervical length of more than 20 mm on transvaginal sonography were independent predictors of Caesarean delivery. This information is helpful for pre-induction counselling. PMID- 17116056 TI - The psychosocial outcomes of antenatal day care for three medical complications of pregnancy: a randomised controlled trial of 395 women. AB - BACKGROUND: Although antenatal day care is becoming increasingly common, there is little evidence as to the psychosocial efficacy of this model of care. AIM: We aimed to assess the broader psychosocial impact of antenatal day care compared with admission to hospital. METHODS: We carried out a randomised trial of 395 women, randomly assigned in a 2 : 1 ratio between day care and antenatal ward, stratified for major diagnostic categories (proteinuric hypertension, non proteinuric hypertension and preterm premature rupture of membranes). Main outcome measures--self-report questionnaires (response rates ranging from 80 to 90%) were sent to women's homes four days after randomisation and seven weeks after delivery. RESULTS: Overall, there were statistically significant differences favouring day care in 12 of 28 items at four days post-randomisation, with no differences in the two groups for the other 16 items. At seven weeks postdelivery, we found differences in eight of 28 items favouring day care, with no differences in the two groups for the other 20 items. The types of items indicating a sustained difference covered a range of aspects of care and included satisfaction with staff, continuity of carer, information transfer, and social support. There were no differences in relation to infant feeding and relationship with the baby. CONCLUSIONS: Day care has an effect on women's satisfaction with care but does not produce broader psychosocial outcomes. PMID- 17116057 TI - Laparoscopic colposuspension versus vaginal suburethral slingplasty: a randomised prospective trial. AB - BACKGROUND: This study aimed to determine if laparoscopic colposuspension (LC) was as effective as vaginal suburethral slingplasty (SPARC). METHODS: Ninety seven women with urodynamic stress incontinence were prospectively randomised to LC (n=48) or SPARC (n=49). Outcome measures were measured at, baseline, six months (n=87) and two years (n=58), and comprised leakage episodes per week and visual analogue scale (VAS) of incontinence severity. The LC and SPARC groups at baseline had similar leaks per week (8.8 vs 9.8) and VAS (5.6 vs 5.9). RESULTS: Laparoscopic colposuspension took longer to perform (48 vs 30 mins, P<0.001), had a slightly higher blood loss (104 vs 82 mL, P<0.01), had a longer hospitalisation (4.0 vs 1.5 days, P<0.001) and had a longer time to resumption of normal activities (3.6 vs 2.8 week, P<0.01). At six months there were no significant differences between LC and SPARC with regard to leaks per week (1.1 vs 2.6) and VAS (1.3 vs 0.7). The success rates were similar (88.3 vs 81.8%). These results again had no significant differences at two years (leaks per week 2.1 vs 3.5, and VAS 1.7 vs 2.2). At two years, the cure/improved rates again found no significant difference (81.5 vs 77.4%) CONCLUSIONS: Laparoscopic colposuspension is as effective as vaginal suburethral slingplasty after two years' follow-up. PMID- 17116058 TI - Risk factors for ectopic pregnancy: a case-control study. AB - AIM: To identify the risk factors for ectopic pregnancy. METHODS: We conducted a prospective case-control study for the role of several risk factors in the occurrence of ectopic pregnancy in Turkey. A total of 225 cases and 375 controls were compared for sociodemographic characteristics, cigarette smoking, obstetric, gynaecological, surgical histories, the presence or absence of assisted conception and contraceptive usage. RESULTS: The main risk factors for ectopic pregnancy were prior ectopic pregnancy (adjusted odds ratio (AOR): 13.1) and a history of infectious reproductive system (AOR for pelvic inflammatory disease: 6.8). Other risk factors found to be associated with an increased risk for ectopic pregnancy were multisexual partner (AOR: 3.5), history of infertility (AOR: 2.5), induced conception cycle (AOR: 3.4), current intrauterine device usage (AOR: 3.2), prior Caesarean section (AOR: 2.1) and cigarette smoking at the time of conception (AOR=1.7). On the contrary, barrier methods were protective from ectopic pregnancy (AOR: 0.4). CONCLUSIONS: The increased awareness and knowledge of risk factors have enabled an early and accurate diagnosis of ectopic pregnancy. This study has found prior pelvic infection to be a major aetiological factor for ectopic pregnancy. Furthermore, other factors found to be associated with ectopic pregnancy, such as prior ectopic pregnancy, infertility history and induced conception cycle, may be the result of a previous pelvic infection that may cause tubal sequelae. These factors are potential targets for intervention and modification. PMID- 17116059 TI - Inadequate compliance with periconceptional folic acid supplementation in South Australia. AB - BACKGROUND: Despite recommendations for women to take folic acid supplements, there has been little reduction in the number of neural tube defect cases occurring each year. AIMS: To assess the level of compliance of pregnant women to recommendations for folic acid supplementation in South Australia, and audit the labelled content in reported supplements used. METHODS: A survey was conducted from May to September 2005 with 304 pregnant women recruited from antenatal clinics at the Lyell McEwin Health Service, the Women's and Children's Hospital and the Modbury Hospital in Adelaide. RESULTS: Full compliance with supplementation recommendations for both timing and dose was achieved by 30% of women. Partial compliance was achieved by 43%, while 27% took no folic acid supplements. CONCLUSIONS: There is currently poor compliance with folic acid supplementation around conception. Additional food fortification may better achieve an adequate daily level of folic acid. PMID- 17116060 TI - Periconceptional folic acid use among women giving birth at Queen Mary Maternity Hospital in Dunedin. AB - BACKGROUND: The New Zealand Ministry of Health advises that all women planning a pregnancy take a folic acid supplement to reduce the risk of having a neural tube defect (NTD)-affected pregnancy. There is little information available to determine if women are following this advice. OBJECTIVE: The purpose of this study was to determine periconceptional folic acid use among women in the postnatal ward of Queen Mary Maternity Hospital in Dunedin. METHODS: A questionnaire was administered to women in the postnatal ward between 14 November and 22 December 2004. RESULTS: One hundred and six women were interviewed during the study period. Forty women (39%) used folic acid supplements before conception. Sixty-seven women (64%) planned their pregnancy. The proportion of women (P<0.001) who planned their pregnancy (53%) and used folic acid before conception was higher than those who did not (11%). The proportion of women 30 years of age (55%) who took folic acid supplements before conception was higher than women aged 17 to <25 years (10%). CONCLUSION: Despite a lack of a public health campaign in New Zealand, a high proportion of participants, especially those who planned their pregnancy, took folic acid during the periconceptional period. A comprehensive public health campaign is needed to increase folic acid use. Fortification may be required to reach those women who do not plan their pregnancies. PMID- 17116061 TI - Postgraduate obstetrics and gynaecology experience: the role for early exposure. AB - BACKGROUND: The early postgraduate years provide the opportunity for junior doctors to gain a range of experience and determine their focused areas of interest. Internship thus requires experience in medicine, surgery and the emergency department. Obstetrics and gynaecology (O & G) is not offered as an internship rotation, despite its suitability as such. A cross-sectional analysis of junior doctors was performed to assess residents' views on the suitability of an O & G rotation for career decision-making and for achieving sufficient clinical experience. METHODS: Fifty-two resident medical staff in their second postgraduate year were recruited and each was given an anonymous survey for completion. Visual analogue scales (VAS) were used for participant responses. RESULTS: For whether internship without an O & G rotation provides sufficient experience to include or exclude O & G as a career choice, the average participant response was 2.1 (on a VAS out of 10) (P<0.0001). For whether an O & G internship rotation would provide adequate medical, surgical or emergency department experience, the average response was 7.9 (on the VAS out of 10) (P<0.0001). As to whether participants would have preferred to undergo an O & G rotation during internship, the average response was 7.7 (P<0.0001). CONCLUSION: The present study demonstrated a perception by residents that an O & G rotation during internship would achieve adequate clinical competencies and assist in career decision-making in O & G, with residents preferring to have undergone an O & G rotation during internship. PMID- 17116062 TI - Uterine rupture: preventable obstetric tragedies? AB - BACKGROUND: Although ruptured uterus is nowadays a rare obstetric emergency in Western countries, it is still alarmingly common in developing countries, where it remains a major cause of maternal mortality and morbidity. AIMS: To review the recent experience of uterine rupture at a tertiary obstetric unit in eastern Nepal and to recommend improvements in the current management of labour, especially obstructed labour, in a poorly resourced country. METHODS: All cases of uterine rupture managed from March 2002 to March 2006 were identified retrospectively, and details were retrieved from medical records. RESULTS: Fifty two women suffered from uterine rupture during the four-year period, approximately one woman per month. Most were unbooked multigravidae, with no antenatal care. They nearly all began labour at home in the absence of a skilled birth attendant. After prolonged labour, usually prolonged second stage, various interventions had often been attempted at home or in other health facilities before admission. Most were shocked and required urgent laparotomy and blood transfusion. Many required intensive care and ventilatory support. Forty-six per cent required hysterectomy and 5.8% subsequently suffered from a urogenital fistula. The maternal mortality rate in this series was 13.5%, and the stillbirth rate was 94.2%. CONCLUSIONS: Unsafe obstetric practices were identified, especially the injudicious use of oxytocic drugs and fundal pressure in prolonged second stage. Several achievable improvements in obstetric care are recommended, particularly aimed at reducing the delay in women reaching emergency obstetric care when labour is prolonged. PMID- 17116063 TI - Care of women at risk of preterm birth: a survey of reported practice in Australia and New Zealand. AB - A mail-out questionnaire on management of women considered at high risk of preterm birth was sent to all members and fellows of the Royal Australian and New Zealand College of Obstetricians and Gynaecologists. While the survey suggested that there is general consensus among practitioners as to what constitutes an increased risk of preterm birth, there is some variation in investigations and treatment currently undertaken. PMID- 17116064 TI - Timing of placental delivery to prevent post-partum haemorrhage: lessons learned from an abandoned randomised clinical trial. AB - It has been recognised that, if the length of the third stage of labour exceeds 30 min, then there is an increased risk of a post-partum haemorrhage. Recent information has suggested that 18 min is the optimal time for removal of the undelivered placenta to prevent a post-partum haemorrhage. A randomised trial comparing 20 vs. 30 min was stopped after an interim analysis because only eight of 1607 patients' placentas had not delivered by 20 min. A third stage of labour that exceeded 10 min was observed to be significantly correlated with an increased risk of post-partum haemorrhage. PMID- 17116065 TI - Perinatal outcomes and macrosomia in a multi-ethnic population of women with type 2 diabetes. AB - We reviewed pregnancy outcomes of women with type 2 diabetes giving birth over a six-year period, comparing the main ethnic groups. Asian women had significantly smaller babies and lower rates of macrosomia as defined by standard growth charts. Other outcomes were similar between the Asian, European and Polynesian women. PMID- 17116066 TI - Use of extracorporeal membrane oxygenation to manage post-partum pulmonary haemorrhage associated with pulmonary hypertension and aberrant right pulmonary artery. PMID- 17116067 TI - Synchronous granulosa cell tumour of the ovary and fallopian tube adenocarcinoma: two rare gynaecological malignancies. PMID- 17116069 TI - Re: use of uterine Doppler in an Australian level II maternity hospital. PMID- 17116072 TI - Suicide for scrutinizers. AB - OBJECTIVES: The aim is to present information which may be useful to those who scrutinize mental health professionals and systems in the aftermath of completed suicide. CONCLUSION: Suicide has been known in all cultures, throughout history, and may be committed in the absence of mental disorder. Risk factors for suicide are known but they are of limited clinical utility. Prediction and prevention of suicide are beyond current abilities. Criticism of mental health professionals and systems in the aftermath of suicide may be destructive and work to the disadvantage of patients. PMID- 17116073 TI - Books reconsidered: Emile Durkheim, Le Suicide. AB - OBJECTIVE: To reappraise Emile Durkheim's taxonomy of suicide in Le Suicide in the light of recent experience. CONCLUSIONS: While Durkheim's scientific method and argument are fundamentally flawed, some of his concepts have instrumental value in attempting to understand the complex origins of suicidal behaviour. Durkheim's baseless dismissal of mental illness as a key determinant of suicidal behaviour weakens his thesis significantly. However, his conceptualization of anomic, egoistic and altruistic suicide provides a means of comprehending recent trends in suicidal behaviour in the former Soviet states and a possible window into the psyche of the suicides of religious and political extremists. PMID- 17116074 TI - The role of child and adolescent mental health services in suicide prevention in New Zealand. AB - OBJECTIVE: We consider the role of child and adolescent mental health services (CAMHS) in the prevention of suicide by young people in New Zealand by examining the conflicting demands placed on CAMHS as a result of public expectation, health care policy and structures, the current focus in the health sector on risk assessment and the inability of CAMHS to see all the young people who may consider suicide in any given year. CONCLUSIONS: We suggest that CAMHS are currently under significant pressures which are inadvertently leading to distortions of clinical practice away from the clinical needs of suicidal children, adolescents and their families towards risk assessment and risk management strategies. The inability of CAMHS to offer comprehensive services to all young people considering suicide may be addressed by greater provision of consult-liaison activities, better understanding between clinicians and health managers, and improvements in clinical skills. PMID- 17116075 TI - Hepatitis C in people with mental illness: how big is the problem and how do we respond? AB - OBJECTIVE: To review the evidence available on best practice care of people with both hepatitis C and mental illness and its application in a mental health service. METHODS: A literature search was conducted for publications dealing with screening, referral for specialist review and antiviral treatment for this population group. RESULTS: A small number of studies was identified that specifically dealt with screening and treatment for hepatitis C in people with mental illness. CONCLUSIONS: Screening, referral and treatment for hepatitis C in people with mental illness is worthwhile and achievable. PMID- 17116076 TI - Prevalence of psychotropic use in a South Australian population. AB - OBJECTIVE: To determine the prevalence of psychotropic medication use in a South Australian population. METHODS: Face-to-face interviews with a random and representative sample of the South Australian population (at least 15 years of age) living in metropolitan and rural areas between March and June 2004. RESULTS: From 4700 households selected, 3015 participants were interviewed (65.9% response rate). Of the respondents, 10.6% were taking psychotropic medications; psychotropic drug use was higher in females (14.4%) than in males (6.8%), and increased with age. Antidepressants were the most common category of psychotropics reported by respondents (6.8%), followed by anxiolytics (1.8%), hypnotics and sedatives (1.5%) and antipsychotics (0.6%). The majority (64.8%) of those respondents who were taking a tricyclic antidepressant were on a dose of 50 mg or less. Only 0.07% of respondents were taking lithium. CONCLUSIONS: The use of psychotropic medications was higher than recently reported studies conducted in the UK, the USA, Europe and Canada. More detailed enquiry about the indication for such use appears desirable in order to ascertain whether higher use in this Australian population is clinically appropriate. PMID- 17116077 TI - Mother-infant psychotherapy and perinatal psychiatry: current clinical practice and future directions. AB - OBJECTIVE: To explore trends in the practice of mother-infant psychotherapy among perinatal psychiatry clinicians based in Melbourne. METHODS: A cross-sectional survey with a purpose designed self-report questionnaire was used to assess the attitudes and practices of 47 perinatal and infant psychiatry clinicians in their use and understanding of mother-infant psychotherapy. RESULTS: Seventy per cent of clinicians in this field of psychotherapy who responded to the questionnaire subscribe to a psychodynamic model, although cognitive behavioural models are also used. The interventions were mostly used in conjunction with other interventions, would be more accurately described as 'parent-infant psychotherapy', and non-psychiatrists in the area tended to be more likely to be formally trained in psychotherapy, but only 4% were formally trained in specific mother-infant psychotherapy. There was a unanimous request for further clinical training in this area. CONCLUSIONS: The emerging field of perinatal psychiatry needs to develop coherent therapeutic models and conduct outcome trials on specific interventions. Specific trainings in these models, in assessment and in diagnostic frameworks are required to enhance clinical efficacy, for research and service development purposes. PMID- 17116078 TI - Grief and courage in a river town: a pilot project in the Aboriginal community of Kempsey, New South Wales. AB - OBJECTIVE: To describe a pilot project implemented in Kempsey, NSW, whereby a psychiatry registrar worked in a community-controlled Aboriginal medical service, Durri Aboriginal Corporation Medical Service, which provides mental health services for Aboriginal people. CONCLUSIONS: The Aboriginal Mental Health Workers and psychiatry registrar jointly managed a significant number of Aboriginal people, often with assistance from the local mental health service at Kempsey. The registrar gained insight into the local Aboriginal community and more culturally appropriate clinical methods, as well as some cultural factors that can influence the presentation and management of mental illnesses. The Aboriginal Mental Health Workers increased their clinical knowledge and confidence through working with the registrar. PMID- 17116079 TI - Do clinical pathways really improve clinical performance in mental health settings? AB - OBJECTIVE: Royal Brisbane and Women's Hospital (RBWH) Mental Health developed and trialled two clinical pathways on psychosis and depression between 2001 and 2003 with the aim of improving a range of clinical and financial parameters. There was a strong commitment from senior management, appropriate resources were allocated and there was adequate staff support. Following a 6-month trial, the pathways were reworked extensively and combined into an acute inpatient pathway. From October 2003 to 2004, we trialled the acute inpatient clinical pathway, and monitored clinical and financial parameters. CONCLUSION: Over this 12-month period, the acute inpatient clinical pathway failed to demonstrate improvement on a range of clinical and financial parameters and its use was ceased. This trial lends support to the view that the complexity, individuality and variability of mental disorders means that clinical pathways are not beneficial in mental health settings. PMID- 17116080 TI - Parenting and mental illness: a pilot group programme for parents. AB - OBJECTIVE: To describe a pilot group programme for parents with a mental illness that was developed and implemented on the NSW Central Coast. CONCLUSIONS: The Parenting and Mental Illness Group Program is a 6-week group programme that is followed by four weekly, individual home visits. Data from the Eyberg Child Behaviour Inventory and Parenting Scale suggest the programme produced positive outcomes in children's behaviour and parenting practices; the programme was also viewed favourably by participants. While encouraging, these findings require replication, using larger numbers and evaluation of the programme in other sites. Supported by an evidence base, programmes such as this will be important to implement if mental health services are to improve outcomes for parents with mental health problems and their children. PMID- 17116081 TI - Psychiatric emergencies in children and adolescents: an Emergency Department audit. AB - OBJECTIVE: To describe a cohort of children presenting to a paediatric emergency department with mental health problems. METHODS: An Emergency Department (ED) computerized record system and hospital records were used to obtain data on children who presented to a paediatric ED with mental health problems. RESULTS: There were 291 presentations of 231 children in a 10-month period, about one per day. They were a small (0.8%) but complicated part of the ED workload. Most were first presentations and came voluntarily to ED. There were a wide variety of presenting symptoms including self-harm, suicide attempts, behavioural disorders and medical disorders with associated psychological problems. Acute psychosis was rare. CONCLUSIONS: Many children with mental health problems were seen for the first time in ED. With the growing awareness of mental health problems in this age group, it is likely that such presentations will increase. Paediatric and psychiatry services have an opportunity to work together to provide early intervention services for what is potentially a very accessible population. PMID- 17116082 TI - Spirituality, religion and psychiatry: its application to clinical practice. AB - OBJECTIVE: Based on evidence obtained from recent Australian psychiatric patient surveys, a need to include the spiritual and religious dimension of patients in their psychiatric care has been identified. This paper aims to review the evidence for this need and to suggest the parameters in which this dimension might be applied. METHODS: The phenomenology of spirituality and its relevance to psychiatry is considered, the concept of the psychiatrist and the clinician as a healer visited, and the evidence for the need for spirituality and religiosity for patients examined. RESULTS: Patients' spiritual needs should be addressed at different levels. Using previous data and experience, the authors suggest what psychiatrists might and might not do, in order that these issues are attended to in an ethical and sensitive manner. CONCLUSIONS: In considering the spiritual dimension of the patient, the psychiatrist is able to send an important message that he or she is concerned with the whole person, a message that enhances the patient-physician relationship. This, in turn, is likely to increase the therapeutic impact of psychiatrists' interventions. PMID- 17116083 TI - Using the Internet to enhance the treatment of depression. AB - OBJECTIVE: Deficiencies in Australia's mental health systems persist despite over a decade of mental health reform. Recent developments in e-health provide the opportunity to facilitate health reform and improve services. This paper presents preliminary findings from the implementation of a comprehensive e-health system, called RecoveryRoad, which was designed to augment the routine clinical treatment of depression. METHODS: Depressed patients (n = 144) were referred to RecoveryRoad from a public hospital and public and private clinics in Perth, Western Australia. Online features included secure e-consultations, progress monitoring questionnaires, psychoeducation and evidence-based therapy. Treating clinicians had online access to patients' progress monitoring outcomes and e consultations. There were two types of adherence reminders: automated email reminders and personalized case management delivered by email and telephone. RESULTS: Adherence to the system was high (from 53% to 84%, depending on the modality of reminder), and self-reported medication adherence was over 90%. Average depression severity declined from severe to mild by the eighth session, a large effect (d = 1.0). Both clinicians and patients were generally satisfied with the programme and reported that it improved clinician-patient relationships. Clinicians also reported that it helped patients to better manage depression. CONCLUSIONS: Preliminary findings support the feasibility of comprehensive e health systems in enhancing the delivery of mental health care. PMID- 17116084 TI - Innocents massacre. PMID- 17116085 TI - Malingered psychosis leading to involuntary psychiatric hospitalization. AB - OBJECTIVE: To describe a patient with feigned psychosis who was admitted to an acute psychiatric unit under the Mental Health Act, and to discuss the clinical features of malingering in this treatment setting. METHOD: Case report. RESULTS: A 40-year-old man with no known past psychiatric history was brought by police to the Emergency Department. He gave a history of paranoia, hallucinations and bizarre behaviour and was admitted to a secure ward. Treatment with parenteral medication was required due to hostile and aggressive behaviour. Collateral history was inconsistent with his reported symptoms and, when confronted with this information, he acknowledged fabricating his symptoms in order to obtain a disability pension. CONCLUSIONS: It is important to consider malingering as a differential diagnosis in patients who present with atypical features and have clear external incentives for their behaviour. With improved health literacy and access to health information, individuals who are marginalized in society and have limited social and occupational opportunities may malinger symptoms for obvious personal gain. Inconsistency between reported symptoms and clinical observations, as well as contradictory collateral information from multiple sources, are helpful tools in diagnosing malingering. It is important for clinicians to identify malingerers early and limit unnecessary admissions. PMID- 17116087 TI - Consultation-liaison psychiatry services to nursing homes. PMID- 17116088 TI - Can investigations supersede neurological examination in the diagnosis of early psychosis? PMID- 17116090 TI - International Classification of Headache Disorders, Second Edition (ICHD-2): current status and future revisions. PMID- 17116091 TI - Psychological treatment of recurrent headache in children and adolescents--a meta analysis. AB - Psychologically based interventions such as relaxation training, biofeedback and cognitive-behavioural therapy are increasingly discussed as options for the treatment of migraine and tension-type headache in children and adolescents. In order to determine the state of evidence regarding the efficacy of these treatments, a meta-analysis of randomized controlled studies was conducted. In a comprehensive literature search including data from 1966 to 2004, 23 studies were found meeting the inclusion criteria. Due to the application of the random effects model, generalization of the results is possible. Specific statistical procedures were used to account for a possible publication bias. Significantly more patients improved to a clinically relevant extent (headache reduction > or =50%) in treatment conditions compared with waiting list conditions (high effect sizes). Long-term stability was also confirmed. The analysed treatments lead to improvement (up to 1 year) in headache status in children and adolescents with primary headache. However, more well-designed studies are needed to support and consolidate the conclusions of this meta-analysis and to compare the effects of psychological treatment with those of prophylactic medical interventions (in migraine), to examine potential differences between treatments, to identify moderators of efficacy and to determine effects of treatment on other health related variables such as quality of life. PMID- 17116092 TI - Headache attributed to infection: observations on the IHS classification (ICHD II). AB - The aim of this study was to revise some topics in the chapter "Headache attributed to infections" in the last International Headache Society (IHS) classification. The authors searched for original studies and reviews about headache associated with infections. A checklist was submitted to 15 neurologists to quantify the relevance, comprehensibility and coherence between definitions, criteria and comments for each paragraph. The following paragraphs were fully discussed: (1) headache attributed to lymphocytic meningitis. This topic, being rather heterogeneous, should be divided into different subgroups; (2) headache attributed to HIV/AIDS. Distinctive features are not specified and diagnostic criteria are rather confusing; and (3) chronic post-infection headache. Diagnostic criteria should be reconsidered as the symptom "pain" is not the main diagnostic criterion. The authors propose the revision of three paragraphs of the new IHS classification to better define the most likely headache profile in specific CNS infections. The authors also underline the need to plan further ad hoc prospective studies. PMID- 17116093 TI - Chronic frequent headache in the general population: prevalence and associated factors. AB - We studied the prevalence and short-term natural course of chronic frequent headache (CFH) in the general population and identified risk factors. In the Netherlands everyone is registered at a single general practice. We sent questionnaires to all persons (n = 21 440) aged 25-55 years, registered at 16 general practices. We compared the characteristics of 177 participants with CFH (>14 headache days/month for >3 months) with 141 participants with infrequent headache (1-4 days/month) and 526 without headache (<1 day/month). The prevalence of CFH was 3.7% [95% confidence interval (CI) 3.4, 4.0]. In 5 months, 12% showed a clinically relevant decrease to <7 days/month. In both headache groups 70% were women vs. 41% in the group without headache. Compared with the group with infrequent headache, the CFH group had more subjects with low educational level [35% vs. 11%; odds ratio (OR) 4.3, 95% CI 2.3, 7.8], medication overuse (62% vs. 3%; OR 38.4, 95% CI 13.8, 106.9), sleeping problems (44% vs. 8%; OR 8.1, 95% CI 3.6, 18.1), a history of head/neck trauma (36% vs. 14%; OR 4.0, 95% CI 2.2, 7.1), high scores on the General Health Questionnaire (62% vs. 34%; OR 2.7, 95% CI 1.3, 3.6) and more smokers (45% vs. 19%; OR 3.1, 95% CI 1.9, 5.3). We conclude that headache frequency fluctuates. CFH is common and associated with overuse of analgesics, psychopathology, smoking, sleeping problems, a history of head/neck trauma and low educational level. Female sex is a risk factor for headache, not for chronification of headache. PMID- 17116094 TI - Chronic frequent headache in the general population: comorbidity and quality of life. AB - We studied the nature and extent of comorbidity of chronic frequent headache (CFH) in the general population and the influence of CFH and comorbidity on quality of life. Subjects with CFH (headache on >14 days/month) were identified in a general health survey. We sent a second questionnaire including questions on comorbidity and quality of life to subjects with CFH and subjects with infrequent headache (IH) (1-4 days/month). We recoded comorbidity by using the Cumulative Illness Rating Scale (CIRS) and measured quality of life with the RAND-36, a Dutch version of Short Form-36. CFH subjects (n = 176) had higher comorbidity scores than the IH subjects (n = 141). Mean CIRS scores were 2.94 for CFH and 1.55 for IH [mean difference 1.40, 95% confidence interval (CI) 0.91, 1.89]. The mean number of categories selected was 1.92 in CFH and 1.10 in IH (mean difference 0.82, 95% CI 0.54, 1.11). Fifty percent of CFH subjects had a comorbidity severity level of at least 2, indicating disorders requiring daily medication, compared with 28% of IH subjects (mean difference 22%, 95% CI 12, 33). CFH subjects had more musculoskeletal, gastrointestinal, psychiatric and endocrine/breast pathology than IH subjects. Quality of life in CFH subjects was lower than that of IH subjects in all domains of the RAND-36. Both headache frequency and CIRS score had a negative influence on all domains. We conclude that patients with CFH have more comorbid disorders than patients with infrequent headaches. Many CFH patients have a comorbid chronic condition requiring daily medication. Both high headache frequency and comorbidity contribute to the low quality of life in these patients. PMID- 17116095 TI - Headache patients in primary care and a tertiary care unit in Zurich, Switzerland. AB - The Headache and Pain Clinic (HPC) is a unit of the Zurich Neurology Department, established in 1966. In the present study demographic features, clinical characteristics and medical management of primary and tertiary care patients were compared in two groups of 181 patients each, seen by general practitioners (GPs) or the HPC, respectively, for primary headaches in 1998. There was a preponderance of women and the socially underprivileged in both samples. Chronic headache was overrepresented in the HPC (44.7%). Loss of work for >2 months was found exclusively in the HPC (9.9%). Of the GP patients, 40% were using triptans and 26.5% in the HPC. One-third of both groups had had complementary and alternative medical treatment. Differences in management strategies reflected differences in headache severity and chronicity. Results indicated that remaining shortcomings of diagnosis and treatment of headache in primary care could be minimized by involving GPs in similar non-commercial studies. PMID- 17116096 TI - Triptans in orgasmic headache. AB - Orgasmic headache (headache associated with sexual activity type 2 according to the International Headache Society classification) is a sudden severe headache which occurs at orgasm. Experiences with triptan therapy are described. Two out of four patients with severe headache continuing for >2 h had a positive response to acute triptan therapy. Two out of three patients using triptans as short-term prophylaxis reported a reliable response on several occasions. Triptans might be a treatment option to shorten orgasmic headache attacks after the diagnosis is clear and, particularly, subarachnoid haemorrhage has been excluded. In patients who chose to predict their sexual activity, short-term prophylaxis with oral triptans 30 min before sexual activity might be a therapeutic option in those not responsive to or not tolerating indomethacin. PMID- 17116097 TI - Testing of variants of the MTHFR and ESR1 genes in 1798 Finnish individuals fails to confirm the association with migraine with aura. AB - Among the few independently replicated genetic associations in migraine are polymorphisms in the methylenetetrahydrofolate reductase (MTHFR) and oestrogen receptor (ESR1) genes. We studied the contribution of these genes to migraine susceptibility by genotyping six MTHFR and 26 ESR1 polymorphisms in 898 unrelated migraine with aura (MA) patients and in 900 unrelated healthy controls. There were no differences in the genotype distributions of the previously migraine associated SNPs C677T (MTHFR) and G2014A (ESR1) between cases and controls (P values 0.83 and 0.55, respectively). Thus, we were not able to replicate the previous findings, although our study had considerable power. However, five of the ESR1 SNPs (rs6557170, rs2347867, rs6557171, rs4870062 and rs1801132) that were in strong linkage disequilibrium were nominally associated with MA (uncorrected P-values 0.007-0.034). These results did not, however, remain significant after taking multiple testing into account. Thus it seems unlikely that the studied genes are involved in migraine susceptibility, at least in this sample. PMID- 17116098 TI - Cost-effectiveness of migraine prevention: the case of topiramate in the UK. AB - The aim of this study was to assess the cost-effectiveness of topiramate vs. no preventive treatment in the UK. Model inputs included baseline migraine frequency, treatment discontinuation and response, preventive and acute medical cost per attack [2005 GBP ( pound)] and gain in health utility. Outcomes included monthly migraines averted, acute and preventive treatment costs and cost per quality-adjusted life year (QALY). Topiramate was associated with 1.8 fewer monthly migraines and a QALY gain of 0.0384. The incremental cost of topiramate vs. no preventive treatment was about 10 UK pounds per migraine averted and 5700 UK pounds per QALY. Results are sensitive to baseline monthly migraine frequency, triptan use rate and the gain in utility. Incorporating savings from reduced work loss (about 36 UK pounds per month) suggests that topiramate would be cost saving compared with no preventive treatment. This analysis suggests that topiramate is a cost-effective treatment for migraine prevention compared with no preventive treatment. PMID- 17116099 TI - Secondary intermedius neuralgia-like pain in a young child. PMID- 17116100 TI - Bath-related headache: the first European case. PMID- 17116102 TI - The fixed combination of acetylsalicylic acid, paracetamol and caffeine. PMID- 17116104 TI - Paranasal sinus pathologies in patients presenting with headache as the primary symptom. PMID- 17116106 TI - Comment on "The use of radiolabelled human serum albumin and SPECT/MRI coregistration to study inflammation in the cavernous sinus of cluster headache patients" by Schuh-Hofer et al. PMID- 17116108 TI - The effect of two publicly funded insurance programs on use of dental services for young children. AB - OBJECTIVE: The purpose of this study is to compare the use of dental services for preschool aged children enrolled in North Carolina Medicaid, a traditional program based on a fee-for-service schedule, and North Carolina Health Choice (NCHC), an State Children's Health Insurance Program (S-CHIP) dental insurance program structured similarly to private insurance. STUDY POPULATION: All children (165,858) 1-5 years of age enrolled in Medicaid and S-CHIP (NCHC) at some time during one study year (October 1, 1999-September 30, 2000). DATA SOURCES/EXTRACTION METHODS: Medicaid and NCHC enrollment and dental claims files were obtained for individual children. STUDY DESIGN: An observational study with a retrospective cohort design. Use of dental services for each child was measured as having at least one dental claim during the outcome period (October 1, 1999 September 30, 2000). Multivariable logistic regression models were developed to compare the effect of two differently administered insurance programs on the use of dental services, controlling for demographic, enrollment, and county characteristics. PRINCIPLE FINDINGS: Children enrolled solely in S-CHIP (NCHC) were 1.6 times more likely (95 percent confidence intervals (CI)=1.50-1.79) to have a dental visit than those enrolled solely in Medicaid. Prediction models for children enrolled for 12 months indicated that those enrolled in S-CHIP (NCHC) had a significantly higher probability of having a dental visit (50 percent) than those enrolled in both plans (44 percent) or Medicaid only (39 percent), a trend found in all age groups. CONCLUSIONS: The S-CHIP (NCHC) program appears to provide children with increased access to dental care compared to children in the Medicaid program. PMID- 17116109 TI - Health insurance take-up by the near-elderly. AB - OBJECTIVE: To examine the effect of price on the demand for health insurance by early retirees between the ages of 55 and 64. DATA SOURCE: Administrative health plan enrollment data from a medium-sized U.S. employer. STUDY DESIGN: The analysis takes advantage of a natural experiment created by the firm's health insurance contribution policy. The amount the firm contributes toward retiree health insurance coverage depends on when a person retired and her years of service at that date. As a result of this policy, there is considerable variation in out-of-pocket premiums faced by individuals in the data. This variation is independent of the nonprice attributes of the health insurance plans offered and is plausibly exogenous to individual characteristics that are likely to affect the demand for insurance. A probit model is used to estimate the decision to take up employer-sponsored health insurance by early retirees between the ages of 55 and 64. Demand for insurance is measured as a function of out-of-pocket premiums and a set of individual characteristics. PRINCIPAL FINDINGS: We find that price has a small but statistically significant effect on the decision to take up coverage. Estimated price elasticities range from -0.10 to -0.16, depending on the sample. CONCLUSIONS: The implied elasticities are comparable with results found in previous studies using very different data. Our estimates indicate that policy proposals for a Medicare buy-in or a nongroup tax credit will have a modest impact on take-up rates of near-elderly retirees. PMID- 17116110 TI - What's driving the downward trend in employer-sponsored health insurance? AB - OBJECTIVE: We investigate the factors driving the downward trend in employer sponsored health insurance (ESI) coverage between 1999 and 2002 for low- and middle-income workers, and assess their insurance options in the absence of ESI coverage. DATA: We use the 1999 and 2002 rounds of the National Survey of America's Families (NSAF), supplemented with ESI premiums from the Medical Expenditure Panel Survey, as well as other state- and county-level data from a variety of sources. The sample includes workers between the ages of 19 and 64. STUDY DESIGN: We first estimate linear probability models of the probability of having an ESI offer and, for those with an offer, the probability of taking up ESI coverage, using two-stage least square regression on the 2002 worker sample. We then use Oaxaca-Blinder regression-based decomposition methods to identify the factors that explain the changes in ESI offer and take-up between 1999 and 2002. PRINCIPAL FINDINGS: We find that while low-income workers are more likely to be uninsured and are most vulnerable to the loss of ESI coverage, many middle-income workers are also in a precarious position when faced with the loss of ESI coverage. Many low- and middle-income workers have few coverage options in the absence of ESI. This is particularly problematic for low-income workers: only 13 percent have a spouse with an ESI offer and the nongroup premium they face increased at a much higher rate than for middle-income workers. Finally, we find that the drop in ESI offers between 1999 and 2002 was driven largely by changes in nature of the workers' jobs, while the drop in ESI take-up was driven largely by rising ESI premiums. CONCLUSIONS: Policies that shore up the ESI system are important for both low- and middle-income workers, as both are vulnerable to a loss of insurance coverage in the absence of ESI. Over time, the potential coverage options available to low- and middle-income workers in the absence of ESI have narrowed as nongroup premiums have increased. While public coverage has provided some protection from that increase for low-income workers, middle-income workers are much less likely to have access to public protection. PMID- 17116111 TI - Estimation of a hedonic pricing model for Medigap insurance. AB - OBJECTIVE: This paper uses a unique database to examine premiums paid by beneficiaries for Medigap supplemental coverage. Average premiums charged by insurers are reported, as well as premiums by enrollee age and gender, and additional policy characteristics. Marginal prices for Medigap benefits are estimated using hedonic price regressions. In addition, the paper considers how additional policy characteristics and geographic differences in the use and cost of medical care affect premiums. DATA SOURCES/STUDY SETTING: A comprehensive database on premiums paid by beneficiaries for newly issued Medigap policies in the year 2000 along with state-level characteristics. STUDY DESIGN: Hedonic pricing equations are used to estimate implicit prices for Medigap benefits. DATA COLLECTION/EXTRACTION METHODS: The Centers for Medicare & Medicaid Services contracted for the creation of a detailed database on Medigap premiums. Data were collected in three stages. First, letters were sent directly to insurers requesting premium data. Second, letters were directly to state insurance commissioner's offices requesting premium data. Last, each state insurance commissioner's office was visited to collect missing data. PRINCIPAL FINDINGS: With the exceptions of the part B deductible and drug benefit, Medigap supplemental insurance is priced consistent with the actuarial value of benefits offered under the standardized plans. Premiums vary substantially based on rating method, whether the policy is guaranteed issue, Medigap Select, or explicitly for smokers. Premiums increase with enrollee age, but do not vary between men and women. The relationship between premiums and enrollee age varies across rating methods. Attained-age policies show the strongest relationship between age and premiums, while community-rated premiums, by definition, do not vary with age. Medigap supplemental insurance premiums are higher in states with poorer health, greater utilization, and greater managed care penetration. CONCLUSIONS: Despite the high cost, Medigap plans are generally priced in accordance with the actuarial value of benefits. The primary exception is the drug benefit, which appears to be subject to substantial adverse selection. Benefits such as the part B deductible and at-home recovery benefit offer little value to consumers. Several states require insurers to community rate premiums. Such regulation has important implications for premiums, and research needs to consider the impact of such regulation on the Medigap market. PMID- 17116112 TI - Is some provider advice on smoking cessation better than no advice? An instrumental variable analysis of the 2001 National Health Interview Survey. AB - RESEARCH OBJECTIVE: To estimate the effect of provider advice in routine clinical contacts on patient smoking cessation outcome. DATA SOURCE: The Sample Adult File from the 2001 National Health Interview Survey. We focus on adult patients who were either current smokers or quit during the last 12 months and had some contact with the health care providers or facilities they most often went to for acute or preventive care. STUDY DESIGN: We estimate a joint model of self reported smoking cessation and ever receiving advice to quit during medical visits in the past 12 months. Because providers are more likely to advise heavier smokers and/or patients already diagnosed with smoking-related conditions, we use provider advice for diet/nutrition and for physical activity reported by the same patient as instrumental variables for smoking cessation advice to mitigate the selection bias. We conduct additional analyses to examine the robustness of our estimate against the various scenarios by which the exclusion restriction of the instrumental variables may fail. PRINCIPAL FINDINGS: Provider advice doubles the chances of success in (self-reported) smoking cessation by their patients. The probability of quitting by the end of the 12-month reference period increased from 6.9 to 14.7 percent, an effect that is of both statistical (p < .001) and clinical significance. CONCLUSIONS: Provider advice delivered in routine practice settings has a substantial effect on the success rate of smoking cessation among smoking patients. Providing advice consistently to all smoking patients, compared with routine care, is more effective than doubling the federal excise tax and, in the longer run, likely to outperform some of the other tobacco control policies such as banning smoking in private workplaces. PMID- 17116113 TI - Colorectal cancer screening in the elderly population: disparities by dual Medicare-Medicaid enrollment status. AB - OBJECTIVES: To assess the disparities in colorectal cancer (CRC) screening between elderly dual Medicare-Medicaid enrollees (or duals), the most vulnerable subgroup of the Medicare population, and nonduals. DATA SOURCES/STUDY SETTING: The 1999 Medicare Denominator File, the Medicare Outpatient Standard Analytic Files, and Physician Supplier Part B files. In addition, the 1998 Area Resource File was used as a source for county-level attributes. DATA COLLECTION/EXTRACTION METHODS: CRC screening procedures for 1999-fecal occult blood test (FOBT), flexible sigmoidoscopy (FLEX), colonoscopy with FOBT and/or FLEX (COL-WFF), and colonoscopy only (COL-ONLY)-were extracted from claim records, using diagnostic and procedure codes. Duals (n = 2.5 million) and nonduals (n = 20.2 million) receiving their care through the fee-for-service system were identified from the Denominator file. Hierarchical logistic regression analysis was conducted to adjust for individual- and county-level characteristics. PRINCIPAL FINDINGS: Compared with nonduals, duals were disproportionately represented by female, older-old, and minority individuals (respectively 74.4 versus 58.5 percent; 19.3 versus 10.8 percent; 35.7 versus 8.0 percent), and CRC screening was significantly lower in duals than in nonduals (5.1 versus 12.2 percent for FOBT adjusted odds ratio [AOR]: 0.48, 95 percent confidence interval [CI]: 0.45-0.51); 0.7 versus 1.9 percent for FLEX, (AOR: 0.55, 95 percent CI: 0.49-0.61); 0.4 versus 0.8 percent for COL-WFF (AOR: 0.60, 95 percent CI: 0.54-0.67); and 1.8 versus 2.5 percent for COL-ONLY (AOR: 0.85, 95 percent CI: 0.80-0.89); p < .001 for all comparisons. CONCLUSIONS: Duals are significantly less likely than nonduals to undergo CRC screening, even after adjusting for individual- and county-level covariates. Future studies should evaluate the contribution of comorbidity and low socioeconomic status to these disparities. PMID- 17116114 TI - Comparing United States versus international medical school graduate physicians who serve African- American and White elderly. AB - OBJECTIVE: To examine the relationship that international medical school graduates (IMGs) in comparison with United States medical school graduates (USMGs) have on health care-seeking behavior and satisfaction with medical care among African-American and white elderly. DATA SOURCES: Secondary data analysis of the 1986-1998 Piedmont Health Survey of the Elderly, Established Populations for the Epidemiological Study of the Elderly, a racially oversampled urban and rural cohort of elders in five North Carolina counties. STUDY DESIGN: Primary focus of analyses examined the impact of the combination of elder race and physician graduate status across time using a linear model for repeated measures analyses and chi2 tests. Separate analyses using generalized estimating equations were conducted for each measure of elder characteristic and health behavior. The analytic cohort included 341 physicians and 3,250 elders (65 years old and older) in 1986; by 1998, 211 physicians and 1,222 elders. DATA COLLECTION/EXTRACTION METHODS: Trained personnel collected baseline measures on 4,162 elders (about 80 percent responses) through 90-minute in-home interviews. PRINCIPAL FINDINGS: Over time, IMGs treated more African-American elders, and those who had less education, lower incomes, less insurance, were in poorer health, and who lived in rural areas. White elders with IMGs delayed care more than those with USMGs. Both races indicated being unsure about where to go for medical care. White elders with IMGs were less satisfied than those with USMGs. Both races had perceptions of IMGs that relate to issues of communication, cultural competency, ageism, and unnecessary expenses. CONCLUSION: IMGs do provide necessary and needed access to medical care for underserved African Americans and rural populations. However, it is unclear whether concerns regarding cultural competency, communication and the quality of care undermine the contribution IMGs make to these populations. PMID- 17116115 TI - How do doctors in different countries manage the same patient? Results of a factorial experiment. AB - OBJECTIVE: To determine the relative contributions of: (1) patient attributes; (2) provider characteristics; and (3) health care systems to health care disparities in the management of coronary heart disease (CHD) and depression. DATA SOURCES/STUDY SETTING: Primary experimental data were collected in 2001-2 from 256 randomly sampled primary care providers in the U.S. (Massachusetts) and the U.K. (Surrey, Southeast London, and the West Midlands). STUDY DESIGN: Two factorial experiments were conducted in which physicians were shown, in random order, two clinically authentic videotapes of "patients" presenting with symptoms strongly suggestive of CHD and depression. "Patient" characteristics (age, gender, race, and socioeconomic status [SES]) were systematically varied, permitting estimation of unconfounded main effects and the interaction of patient, provider, and system-level influences. DATA COLLECTION/DATA EXTRACTION METHODS: Analysis of variance was used to measure provider decision-making outcomes, including diagnosis, information seeking, test ordering, prescribing behavior, lifestyle recommendations, and referrals/follow-ups. PRINCIPAL FINDINGS: There is a high level of consistency in decision making for CHD and depression between the U.S. and the U.K. Most physicians in both countries correctly identified conditions depicted in the vignettes, although U.S. doctors engage in more information seeking, are more likely to prescribe medications, and are more certain of their diagnoses than their U.K. counterparts. The absence of any national differences in test ordering is consistent for both of the medical conditions depicted. U.K. physicians, however, were more likely than U.S. physicians to make lifestyle recommendations for CHD and to refer those patients to other providers. CONCLUSIONS: Substantively, these findings point to the importance of patient and provider characteristics in understanding between country differences in clinical decision making. Methodologically, our use of a factorial experiment highlights the potential of these methods for health services research-especially the estimation of the influence of patient attributes, provider characteristics, and between-country differences in the quality of medical care. PMID- 17116116 TI - Variation in chemotherapy utilization in ovarian cancer: the relative contribution of geography. AB - OBJECTIVE: This study investigates geographic variation in chemotherapy utilization for ovarian cancer in both absolute and relative terms and examines area characteristics associated with this variation. DATA SOURCES: Surveillance, Epidemiology, and End Results (SEER) Medicare data from 1990 to 2001 for Medicare patients over 65 with a diagnosis of ovarian cancer between 1990 and 1999. Chemotherapy within a year of diagnosis was identified by Medicare billing codes. The hospital referral region (HRR) represents the geographic unit of analysis. STUDY DESIGN: A logit model predicting the probability of receiving chemotherapy by each of the 39 HRRs. Control variables included medical characteristics (patient age, stage, year of diagnosis, and comorbidities) and socioeconomic characteristics (race, income, and education). The variation among HRRs was tested by the chi2 statistic, and the relative contribution was measured by the omega statistic. HHR market characteristic are then used to explain HRR-level variation. PRINCIPAL FINDINGS: The average chemotherapy rate was 56.6 percent, with a range by HRR from 33 percent to 67 percent. There were large and significant differences in chemotherapy use between HRRs, reflected by a chi2 for HRR of 146 (df = 38, p < .001). HRR-level variation in chemotherapy use can be partially explained by higher chemotherapy rates in HRRs with a higher percentage of hospitals with oncology services. However, an omega analysis indicates that, by about 15 to one, the variation between patients in use of chemotherapy reflects variations in patient characteristics rather than unexplained variation among HRRs. CONCLUSIONS: While absolute levels of chemotherapy variation between geographic areas are large and statistically significant, this analysis suggests that the role of geography in determining who gets chemotherapy is small relative to individual medical characteristics. Nevertheless, while variation by medical characteristics can be medically justified, the same cannot be said for geographic variation. Our finding that density of oncology hospitals predicts chemotherapy use suggests that provider supply is positively correlated with geographic variation. PMID- 17116117 TI - Racial variation in end-of-life intensive care use: a race or hospital effect? AB - OBJECTIVE: To determine if racial and ethnic variations exist in intensive care (ICU) use during terminal hospitalizations, and, if variations do exist, to determine whether they can be explained by systematic differences in hospital utilization by race/ethnicity. DATA SOURCE: 1999 hospital discharge data from all nonfederal hospitals in Florida, Massachusetts, New Jersey, New York, and Virginia. DESIGN: We identified all terminal admissions (N = 192,705) among adults. We calculated crude rates of ICU use among non-Hispanic whites, blacks, Hispanics, and those with "other" race/ethnicity. We performed multivariable logistic regression on ICU use, with and without adjustment for clustering of patients within hospitals, to calculate adjusted differences in ICU use and by race/ethnicity. We explored both a random-effects (RE) and fixed-effect (FE) specification to adjust for hospital-level clustering. DATA COLLECTION: The data were collected by each state. PRINCIPAL FINDINGS: ICU use during the terminal hospitalization was highest among nonwhites, varying from 64.4 percent among Hispanics to 57.5 percent among whites. Compared to white women, the risk adjusted odds of ICU use was higher for white men and for nonwhites of both sexes (odds ratios [ORs] and 95 percent confidence intervals: white men = 1.16 (1.14 1.19), black men = 1.35 (1.17-1.56), Hispanic men = 1.52 (1.27-1.82), black women = 1.31 (1.25-1.37), Hispanic women =1.53 (1.43-1.63)). Additional adjustment for within-hospital clustering of patients using the RE model did not change the estimate for white men, but markedly attenuated observed differences for blacks (OR for men =1.12 (0.96-1.31), women = 1.10 (1.03-1.17)) and Hispanics (OR for men =1.19 (1.00-1.42), women = 1.18 (1.09-1.27)). Results from the FE model were similar to the RE model (OR for black men = 1.10 (0.95-1.28), black women = 1.07 (1.02-1.13) Hispanic men = 1.17 (0.96-1.42), and Hispanic women = 1.14 (1.06 1.24)) CONCLUSIONS: The majority of observed differences in terminal ICU use among blacks and Hispanics were attributable to their use of hospitals with higher ICU use rather than to racial differences in ICU use within the same hospital. PMID- 17116118 TI - Can the quality of care in family practice be measured using administrative data? AB - OBJECTIVE: To explore the feasibility of using administrative data to develop process indicators for measuring quality in primary care. DATA SOURCES/STUDY SETTING: The Population Health Research Data Repository (Repository) housed at the Manitoba Centre for Health Policy which includes physician claims, hospital discharge abstracts, pharmaceutical use (Drug Program Information Network (DPIN)), and the Manitoba Immunization Monitoring Program (MIMS) for all residents of Manitoba, Canada who used the health care system during the 2001/02 fiscal year. Family physicians were identified from the Physician Resource Database. Indicators were developed based on a literature review and focus group validation. DATA COLLECTION/EXTRACTION METHODS: Data files were extracted from administrative data available in the Repository. We extracted data based on the ICD-9-CM codes and ATC-class drugs prescribed and then linked them to the Physician Resource Database. Physician practices were defined by allocating patients to their most responsible physician. Every family physician in Manitoba that met the inclusion criteria (having either 5 or 10 eligible patients depending on the indicator) was 'scored' on each indicator. Physicians were then grouped according to the proportion of the patients allocated to their practice who received the recommended care for the specific indicator. PRINCIPAL FINDINGS: Using administrative health data we were able to develop and measure eight indicators of quality of care covering both preventive care services and chronic disease management. The number of eligible physicians and patients varied for each indicator as did the percent of patients with recommended care, per physician. For example, the childhood immunization indicator included 544 physicians who, on average, provided immunization for 65 percent of their patients. CONCLUSIONS: Quality of care provided by family physicians can be measured using administrative data. Despite the limitations addressed in this paper, this work establishes a practical methodology to measure quality of care provided by family physicians that can be used for quality improvement initiatives. PMID- 17116119 TI - Estimating the effect of smoking cessation on weight gain: an instrumental variable approach. AB - OBJECTIVE: To propose and test a method that produces an unbiased estimate of the average effect of smoking cessation on weight gain. Previous estimates may be biased due to unobservable differences in attributes of quitters and continuing smokers. An accurate estimate of weight gain due to cessation is important for policymakers, health managers, clinicians, consumers, and developers of smoking cessation aids. STUDY SETTING: Our analysis consisted of an instrumental variables (IVs) approach in which treatment assignment in randomized smoking cessation trials served as a random source of variation in probability of quitting. DATA COLLECTION: We searched the medical literature for previously conducted smoking cessation trials that contained data suitable for our reanalysis. PRINCIPAL FINDINGS: We identified one trial for our reanalysis, the Lung Health Study, a randomized smoking cessation trial with 5,887 smokers aged 35-60 from 1986 to 1994 in several sites across the United States. In our IV reanalysis, we estimated a 9.7 kg weight gain over 5 years due to cessation, as compared with the conventional estimate of 5.3 kg. CONCLUSIONS: The true effect of smoking cessation on weight gain may be larger than previously estimated. This result indicates the importance of fully understanding the possible weight effects of cessation and underscores the need to accompany cessation programs with weight management interventions. The result, however, does not overturn the conclusion that the net health benefits of quitting are positive and very large. The application of the IV technique we propose is likely to be useful in a variety of contexts in which one is interested in the effect of one health condition on another. PMID- 17116120 TI - Effect of dialysis center profit-status on patient survival: a comparison of risk adjustment and instrumental variable approaches. AB - OBJECTIVE: To compare the estimated effects of dialysis center profit status on patient survival using alternative estimation strategies with retrospective data. DATA SOURCES/STUDY SETTING: Patient and provider-level retrospective data from the United States Renal Data System (USRDS), 1996-1999. STUDY DESIGN: Observational risk adjustment and instrumental variable methods. DATA COLLECTION/EXTRACTION METHODS: Study collected measures from various USRDS files describing clinical characteristics, survival, and the profit status of the initial dialysis center for incident end-stage renal disease (ESRD) patients aged 67+. USRDS facility files were used to assess dialysis center profit status and measure patient distances to dialysis centers. PRINCIPAL FINDINGS: Found survival effect related to profit status in the range of previous research using risk adjusting covariates similar to those used in previous models. Adding further risk-adjusting covariates halved this effect. The relative proximity of for profit and nonprofit dialysis centers to the patient residence was the strongest determinant of the profit status of the patient's initial dialysis center. The effect of profit status on survival was eliminated using the two-stage least squares variant of instrumental variable estimation with the relative proximity of for-profit and nonprofit dialysis centers to the patient's residence as the instrument. CONCLUSIONS: Using only the variation in initial dialysis center profit status that was related to the relative proximity of for-profit and nonprofit dialysis centers to the patient, we found no relationship between dialysis center profit status and patient survival. These results are in contrast to results obtained using risk-adjustment methods with a limited set of risk adjusting covariates. PMID- 17116121 TI - Caught in the act? Prevalence, predictors, and consequences of physician detection of unannounced standardized patients. AB - Objective. To examine the prevalence, predictors, and consequences of physician detection of unannounced standardized patients (SPs) in a study of the impact of direct-to-consumer advertising on treatment for depression. Data Sources. Eighteen trained SPs were randomly assigned to conduct 298 unannounced audio recorded visits with 152 primary care physicians in three U.S. cities between May 2003 and May 2004. Study Design. Randomized controlled trial using SPs. SPs portrayed six roles, created by crossing two clinical conditions (major depression or adjustment disorder) with three medication request scripts (brand specific request, general request for an antidepressant, or no request). Data Collection. Within 2 weeks following the visit, physicians completed a form asking whether they "suspected" conducting an office visit with an SP during the past 2 weeks; 296 (99 percent) detection forms were returned. Physicians provided contextual data, a Clinician Background Questionnaire. SPs filled in a Standardized Patient Reporting Form for each visit and returned all written prescriptions and medication samples to the laboratory. Principal Findings. Depending on the definition, detection rates ranged from 5 percent (unambiguous detection) to 23.6 percent (any degree of suspicion) of SP visits. In 12.8 percent of encounters, physicians accurately detected the SP before or during the visit but they only rarely believed their suspicions affected their clinical behavior. In random effects logistic regression analyses controlling for role, actor, physician, and practice factors, suspected visits occurred less frequently in HMO settings than in solo practice settings (p<.05). Physicians more frequently referred SPs to mental health professionals when visits aroused high suspicion (p<.05). Conclusions. Trained actors portrayed patient roles conveying mood disorders at low levels of detection. There was some evidence for differential treatment of detected standardized patients by physicians with regard to referrals but not antidepressant prescribing or follow-up recommendations. Systematic assessment of detection is recommended when SPs are used in studies of clinical process and quality of care. PMID- 17116123 TI - Oral rabeprazole vs. intravenous pantoprazole: a comparison of the effect on intragastric pH in healthy subjects. AB - BACKGROUND: Intravenous pantoprazole is often administered inappropriately to hospitalized patients who can take oral medications. AIM: To compare the antisecretory effects of oral rabeprazole and intravenous pantoprazole in healthy subjects. METHODS: In a double-blind, double-dummy, two-way crossover study, 38 Helicobacter pylori-negative volunteers were randomized to oral rabeprazole 20 mg or intravenous pantoprazole 40 mg daily for 3 days followed, after a 14-day washout period by the comparator treatment. Intragastric pH was recorded continuously for 24 h at baseline and on days 1 and 3 of each treatment period. RESULTS: The mean (95% CI) percentage of the 24-h recording with gastric pH >4 was higher with rabeprazole than with pantoprazole on day 1: 37.7% (30.6-44.8%) vs. 23.9% (20.0-27.8). The mean percentage times with pH >3 and >4 for all intervals assessed were greater and the median 24-h intragastric pH values were higher with rabeprazole than with pantoprazole on days 1 and 3. The mean acidity index was lower with rabeprazole on days 1 and 3. CONCLUSIONS: Oral rabeprazole 20 mg produced greater acid suppression than intravenous pantoprazole 40 mg. Therefore, it may be an appropriate and effective alternative in patients who can take oral medication. PMID- 17116124 TI - Substitution of phenylephrine for pseudoephedrine as a nasal decongeststant. An illogical way to control methamphetamine abuse. AB - The aim of this review was to investigate the rationale for replacing the nasal decongestant pseudoephedrine (PDE) with phenylephrine (PE) as a means of controlling the illicit production of methamphetamine. A literature search was conducted in electronic databases and use of textbooks. Restrictions have been placed on the sale of PDE in the USA in an attempt to control the illicit production of methamphetamine. This has caused a switch from PDE to PE in many common cold and cough medicines. PE is a poor substitute for PDE as an orally administered decongestant as it is extensively metabolized in the gut and its efficacy as a decongestant is unproven. Both PDE and PE have a good safety record, but the efficacy of PDE as a nasal decongestant is supported by clinical trials. Studies in the USA indicate that restricting the sale of PDE to the public as a medicine has had little impact on the morbidity and number of arrests associated with methamphetamine abuse. Restricting the sale of PDE in order to control the illicit production of methamphetamine will deprive the public of a safe and effective nasal decongestant and force the pharmaceutical industry to replace PDE with PE, which may be an ineffective decongestant. Restrictions on sales of PDE to the public may not reduce the problems associated with methamphetamine abuse. PMID- 17116126 TI - Clinical risk management of herb-drug interactions. AB - The concomitant use of conventional and herbal medicines can lead to clinically relevant herb-drug interactions. Clinical risk management offers a systematic approach to minimize the untoward consequences of these interactions by paying attention to: (i) risk identification and assessment; (ii) development and execution of risk reduction strategies; and (iii) evaluation of risk reduction strategies. This paper reviews which steps should be explored or taken in these domains to improve the clinical risk management of adverse herb-drug interactions. PMID- 17116127 TI - Trinucleotide repeat dynamic mutation identifying susceptibility in familial and sporadic chronic lymphocytic leukaemia. AB - Chronic lymphocytic leukaemia (CLL) has a strong hereditary component, but an understanding of predisposition genes is poor. Anticipation with familial CLL has been reported, although the molecular mechanism is unknown. Expansion of trinucleotide repeat sequences underlies anticipation observed in neurodegenerative disease. A polymerase chain reaction-based assay was used to analyse the stability of ten CCG- and CAG-trinucleotide repeat tracts in 18 CLL families and 140 patients with the sporadic form of the disease. The study suggests that anticipation, if it occurs in CLL, is not linked to CCG- and CAG repeat expansion, however, variation in repeat length at certain loci (FRA16A) may permit identification of susceptible family members. In addition, polymorphisms with prognostic significance were identified. These were high length (but not expanded) repeats at FRA11B (P = 0.01), ATXN1 (P = 0.032) and ATXN3 (P = 0.022), all associated with poor risk disease. PMID- 17116128 TI - Safety indicators for inpatient and outpatient oral anticoagulant care: [corrected] Recommendations from the British Committee for Standards in Haematology and National Patient Safety Agency. PMID- 17116129 TI - Combination chemotherapy followed by autologous stem cell transplant for enteropathy-associated T cell lymphoma. AB - Enteropathy-associated T cell lymphoma (EATL) is a rare entity associated with coeliac disease, with a poor prognosis due to perforation and gastro-intestinal bleeding during treatment, and a high relapse risk. Six patients were treated with two cycles of IVE (ifosphamide, etoposide, epirubicin), followed by two cycles of high-dose methotrexate (3 g/m(2)) with folinic acid rescue and a BEAM (carmustine, etoposide, cytarabine, melphalan) autograft. Enteral feeding was given throughout treatment. Four patients remain alive in complete remission at 1.83-4.32 years; two have relapsed. Given the historically poor outcome in these patients, this regimen appears very promising in the treatment of EATL. PMID- 17116130 TI - Validation study of the prediction system for clinical response of M-VAC neoadjuvant chemotherapy. AB - To predict the efficacy of the M-VAC neoadjuvant chemotherapy for invasive bladder cancers, we previously established the method to calculate the prediction score on the basis of expression profiles of 14 predictive genes. This scoring system had clearly distinguished the responder group from the non-responder group. To further validate the clinical significance of the system, we applied it to 22 additional cases of bladder cancer patients and found that the scoring system correctly predicted clinical response for 19 of the 22 test cases. The group of patients with positive predictive scores had significantly longer survival than that with negative scores. When we compared our results with a previous report describing the prognosis of the patients with cystectomy alone, the results imply that patients with positive scores are likely to benefit from M VAC neoadjuvant chemotherapy, but that the chemotherapy would shorten the lives of patients with negative scores. We are confident that our prediction system to M-VAC therapy should provide opportunities for achieving better prognosis and improving the quality of life of patients. Taken together, our data suggest that the goal of 'personalized medicine', prescribing the appropriate treatment regimen for each patient, may be achievable by selecting specific sets of genes for their predictive values according to the approach shown here. PMID- 17116131 TI - Antifungal prophylaxis in transplant recipients: where do we go from here? PMID- 17116132 TI - Antifungal prophylaxis following reduced-intensity stem cell transplantation. AB - Reduced-intensity stem cell transplantation (RIST) has been developed to be a novel curative option for advanced hematologic diseases. Its minimal toxicity allows for transplantation in patients with advanced age or with organ dysfunction. Young patients without comorbidity can undergo RIST as outpatients. However, fungal infection remains an important complication in RIST. Given the poor prognosis of fungal infection, prophylaxis is critical in its management. The prophylactic strategy is recently changing with the development of RIST. Hospital equipment is important for fungal prophylaxis; however, the median day for the development of fungal infection is day 100, when most RIST patients are followed as outpatients. The focus of fungal management after RIST needs to shift from in-hospital equipment to oral antifungals. Various antifungals have recently been developed and introduced for clinical use. A major change in antifungal management will probably occur within several years. PMID- 17116133 TI - Cytomegalovirus infection in renal transplant recipients. AB - We have retrospectively analyzed the incidence of cytomegalovirus (CMV) infection in 250 consecutive renal allograft transplants performed in 244 recipients. The mean follow-up was 35.1+/-25.4 months. Immunosuppression was cyclosporine- or tacrolimus-based triple therapy. CMV infection prophylaxis with ganciclovir for 3 months post transplant was prescribed in CMV-seronegative recipients of allografts from seropositive donors (D+R-) and in all recipients treated with OKT3. CMV antigenemia was monitored by the pp65-antigen assay. Thirteen of 57 D+R recipients (22.8%) developed CMV antigenemia. One recipient had a breakthrough of CMV antigenemia during ganciclovir prophylaxis; 12 D+R- recipients developed CMV antigenemia 147.5+/-173.8 days after transplantation. Four of 13 (30.7%) D+R- recipients had asymptomatic CMV infection, 8 (61.6%) had CMV infection with non specific symptoms including fever, and 1 (7.7%) developed CMV pneumonia. Six of 13 (46.1%) D+R- patients had been treated with intensified immunosuppressive therapy before CMV infection. In the low-risk CMV groups (D+R+; D-R+; D-R-), 28 recipients (14.5%) developed CMV antigenemia 42.5+/-15.2 days post transplantation. Ten of the 28 (35.7%) recipients had asymptomatic CMV infection, 17 (60.7%) developed CMV infection with non-specific symptoms, and 1 (3.6%) developed CMV pneumonia. Twenty-one of 28 (75.0%) had intensified immunosuppressive therapy before CMV infection. In conclusion, ganciclovir prophylaxis diminished and delayed the onset of CMV infection but did not totally prevent it from occurring in D+R- renal transplant recipients. Clinicians should be vigilant to the possibility of CMV infection in both seronegative and seropositive recipients, especially after anti-rejection therapy. PMID- 17116134 TI - Variation in antifungal prophylaxis strategies in lung transplantation. AB - We conducted a survey of 50 lung transplant centers across the world to evaluate the variation in antifungal prophylaxis practices. These 50 centers performed 63% of the world's lung transplants reported in 2001. Eighty-six percent (43/50) of the centers responded to the survey. Sixty-nine percent (30/43) of centers used universal antifungal prophylaxis. Aerosolized amphotericin B deoxycholate (AmBd) alone or in combination with itraconazole was used at 56% (24/43) of centers. The median duration of prophylaxis with aerosolized AmBd and itraconazole was 30 and 90 days, respectively. Seventy-four percent of the centers surveyed agreed to participate in future research prophylaxis protocols, which they felt should include both diagnostic and therapeutic arms. Our survey is the first documentation of the international variation in antifungal prophylactic strategies in lung transplant recipients, and underscores the need for multicenter, randomized trials of antifungal prophylaxis in lung transplant recipients. PMID- 17116135 TI - Elimination of false-positive Histoplasma antigenemia caused by human anti-rabbit antibodies in the second-generation Histoplasma antigen assay. AB - False-positive Histoplasma antigenemia was reported in solid organ allograft recipients who had received rabbit anti-thymocyte globulin (RATG, RATG) caused by human anti-rabbit antibodies (HARA). A second-generation Histoplasma antigen detection assay was developed to overcome false positivity caused by HARA. With the second-generation assay, false-positive results were eliminated in 18 of 19 cases without reduction in the sensitivity in patients with histoplasmosis. In fact, sensitivity for detection of antigenuria in patients with acquired immunodeficiency syndrome and disseminated histoplasmosis was higher in the second-generation assay. Physicians should be aware of the potential for false positive results in sandwich immunoassays in specimens from patients who have received RATG. PMID- 17116136 TI - Successful treatment of disseminated Nocardia farcinica infection in a living donor liver transplantation recipient. AB - Nocardiosis is a serious infection with high mortality. We report a case of subcutaneous and neural lesions due to Nocardia farcinica infection after living donor liver transplantation. The neural lesion was cured with antibiotics without drainage. PMID- 17116137 TI - Colitis in a renal transplant patient with human herpesvirus-6 infection. AB - A male patient developed colitis and a thrombotic microangiopathy 3 weeks after renal transplantation. Immunosuppression at the time of presentation was with sirolimus, mycophenolate mofetil, and prednisolone, but without a calcineurin inhibitor. Cytomegalovirus infection was excluded. However, human herpesvirus-6 DNA was detected at high copy number in both blood and colonic epithelium. The patient recovered after reduction in immunosuppression, with nutritional support and ganciclovir therapy. PMID- 17116138 TI - Primary herpes simplex virus type-2 infection as a cause of liver failure after liver transplantation. AB - We report a case of fatal primary herpes simplex virus type-2 (HSV-2) infection following liver transplantation, which manifested with fever and liver failure in the absence of muco-cutaneous disease. The infection was characterized by high levels of HSV DNA in blood and the patient's inability to mount HSV-specific T cell responses while showing preserved T-cell responses against cytomegalovirus. The donor was HSV-1 immunoglobulin G (IgG) seronegative and HSV-2 IgG seropositive, whereas the recipient was HSV-1 and HSV-2 IgG seronegative, suggesting that the graft may have been the source of the infection. In HSV seronegative recipients of grafts from HSV-seropositive donors, HSV infection should be included in the differential diagnosis of a febrile illness, regardless of the absence of muco-cutaneous disease. In this setting, real-time polymerase chain reaction applied to blood samples provides a sensitive, rapid, and quantitative diagnostic tool. PMID- 17116139 TI - Acute infection of Toxoplasma gondii and cytomegalovirus reactivation in a pediatric patient receiving liver transplant. AB - A 7-year-old Mexican boy with end-stage cirrhosis underwent liver transplantation and was maintained with cyclosporine and prednisolone. No specific data about Toxoplasma gondii or cytomegalovirus (CMV) infections in the cadaver donor were available. The recipient was seronegative for Toxoplasma, but CMV-IgG positive before transplantation. Ganciclovir was administered for prophylaxis during 3 months, but 5 months later he presented with icterus and increased transaminases. Acute transplant rejection was ruled out by biopsy. A seroconversion for T. gondii IgM and IgG and a small increase in CMV-IgM antibodies were observed, although the CMV-polymerase chain reaction (PCR) was negative. Ganciclovir was re started, and the patient improved, but 6 months later he relapsed, and chorioretinitis lesions compatible both with T. gondii and CMV infections appeared. Pyrimethamine, sulfadiazine, folinic acid, and ganciclovir were administered. The boy showed favorable clinical improvement and remained stable for 12 months. Then, new retinal CMV lesions appeared in both eyes and the PCR for CMV became positive; therefore, the patient received a new regimen of ganciclovir, and clinically improved. From these data we concluded that the child presented a reactivation of CMV and a primary infection with T. gondii after transplantation. PMID- 17116140 TI - Non-fatal gastric mucormycosis in a renal transplant patient and review of the literature. AB - Mucormycosis is historically associated with substantial morbidity and carries a very high mortality in renal transplant patients. The common sites of involvement include cerebral, pulmonary, gastrointestinal, or disseminated lesions. We report the successful outcome of gastric mucormycosis along with Strongyloides stercoralis infection in a renal transplant patient, who has survived for 5 years. We also review the literature. PMID- 17116141 TI - Isoniazid prophylaxis in end-stage renal disease. PMID- 17116145 TI - To cope with uncertainty: stroke patients' use of temporal models in narratives. AB - Stroke victims have to cope with a disrupted autobiography and anxiety because of an uncertain future. Professionals share this uncertainty. The patients reveal their experiences in narratives, and when they try to regain coherence and confidence in life, they use narratives in the reconstructions. Because they have a temporal problem, time might be an important issue in these narratives. The aim of this study was to elucidate the use of time models in stroke patients' narratives. Nineteen stroke patients, who had recently been discharged to their homes after the stroke, accepted to participate in the study. Their age span was between 56 and 89 years. They had lived active urban lives before the stroke, and poststroke only three had more serious physical impairment, and none was demented. They were asked to talk about their present life and their conceptions of future life. The interviews were audio-taped and transcribed verbatim and narratives that referred to temporal aspects were thematically analysed with reference to narrative time models. The stroke accident had caused an autobiographical disruption and a temporal split because of a new awareness of human temporality and an uncertainty of the future. Confronted with these problems of time, the stroke victims constructed narratives based on the time models: time cycles and dissolution of time limits, exchange of time and exclusion from time. Hence, the time models worked as tools when the stroke victims re-established coherence in their present and future life. Stroke patients handled an uncertain future by using temporal models in their narratives. Professionals can support stroke patients by reinforcing these models. PMID- 17116146 TI - Consistency of quality assessments in long-term care by the clients, family members and named nurses. AB - Combining assessments by the clients, their family members and named nurses of the quality of long-term care leads to a more comprehensive picture of the quality of the service. Clients should be the primary source of information, but particularly when they are not able to express their opinion, other sources of assessment are needed. This study describes and compares the consistency of quality assessments of long-term institutional care for the elderly made by the client (n = 312), his/her family member (n = 312) and the named nurse (n = 312). Data were gathered in 2002 from service houses, nursing homes and health centres. The consistency of the assessments made by the different groups of respondents was measured by frequency distributions, the weighted kappa coefficient and exact agreement. Frequency distributions differed significantly between the respondent groups in almost every item. Family members were more critical in their assessments than the other two groups. The consistency of assessments was highest with regard to the items 'contact with significant others' and 'medication'. In general the consistency of assessments, measured by weighted kappa, was rather low, but it was fair for 'clothing' (clients/family members) and 'privacy' (clients/named nurses and family members/named nurses). The occurrence of the option 'not applicable' to certain items was quite high, but varied somewhat between the three respondent groups. The results of this study support the viewpoint that comprehensive information gathering from family members and named nurses is useful, but they should not replace clients' assessments. PMID- 17116147 TI - 'It's everywhere!' young Swedish people's thoughts and reflections about pornography. AB - Pornography is one of the most sought-after topics on the Internet, and is easily available for anyone, including children and adolescents. At youth centres, nurse midwives have noticed that young people have different kinds of questions about sexual practices compared with a few years ago. The aim of this study was to gain an understanding of thoughts and reflections about pornography consumption, and its possible influence on sexual practices, among young women and men. The staff at a youth centre in a city in central Sweden asked the visitors if they had seen pornography and if they wanted to be interviewed about their experiences. Ten young women and eight men, aged 16-23 years, participated. In-depth interviews were performed and open-ended questions about pornography and sexuality were posed. The interviews were tape-recorded and transcribed verbatim. Data were analysed according to grounded theory. The core category 'Living with the current sexual norm' depicted how pornography created sexual expectations and demands, for instance, to perform certain sexual acts. The informants expressed contradictory feelings towards pornography and felt that sexuality was separated from intimacy. A moral attitude was described and examples of stereotypic gender roles were given. To deal with the current sexual norm, informants had different individual handling strategies and attitudes to pornography, namely liberal, normalization, distance, feminist or conservative. Limitations of this study were the small sample size and that results from a qualitative research study cannot be generalized. The results contribute to an understanding of how pornographic material can influence young peoples' thoughts, reflections and sexual behaviour. This indicates the importance, for personnel at youth centres and schools, to discuss sexual behaviour and how sexuality is portrayed in pornographic material with young people. PMID- 17116148 TI - Nursing advocacy in an Australian multidisciplinary context: findings on medico centrism. AB - As a follow-up to a recent study which highlighted the existence of medical dominance in multi-disciplinary team (MDT) meetings, this paper presents research findings from an Australian study which shows that medico-centrism is a key cause of tension within MDTs. The findings are from a 1-year qualitative study in a regional hospital that explored the ethical decision-making of health professionals within an acute care medical unit. This exploration was conducted through an iterative, phenomenological, qualitative research methodology that consisted of open-ended interviews with a multi-disciplinary representation of health professionals and a sample of consumers for whom they care. The paper situates the notion of nursing advocacy within the context of medico-centrism and examines how the nursing profession interfaces with other disciplines. The findings indicate that the professional framework of nursing includes the language of advocacy, whilst the framework of doctors centres around the medical decision-making process. All professional groups made reference to the MDT as the modus operandi for patient-centred care. All participants noted that time and familiarity with patients and their families is essential for patient-centred care and this could be achieved through MDT collaboration. However, doctors who have scant time to spend with patients saw it as their responsibility to direct the decisions of the MDT and viewed the MDT as adding confusion to the decision making process. Nurses reported that the limited amount of time spent by doctors in patient consultation translated into the need for advocacy. Professional and clinical confidence and experience are noted as necessary to successfully engage in the process of advocacy. The findings of this article indicate that the adoption of an advocacy role by nurses represents an important means through which MDT operation can be enhanced, medico-centrism limited and patient-centred care improved. PMID- 17116149 TI - Significance of fellow patients for patients with myocardial infarction. AB - The purpose of the study was to present both positive and negative experiences with respect to the significance of fellow patients for patients with myocardial infarction (MI), both inside and outside the hospital. Five focus group sessions were carried out, each having between four and six participants. A total of 25 patients with MI at an age of 40-71 participated. The analysis was carried out by the moderator and co-moderator of the focus groups. The results, which are elucidated by theories of social support, show a great amount of positive support in the interactions of patients, such as support from others in the same situation, a lot of humour, encountering true understanding and consideration, getting practical assistance, and benefiting from other patients' knowledge and experience as well as experiencing an increase in motivation. The participants also discussed negative experiences such as dramatic situations when fellow patients got worse and sometimes even died. Many patients were bothered by various kinds of noise and other disturbances. Insight into the significance of fellow patients will make health personnel able to encourage a patient environment that will strengthen positive and reduce negative effects of fellow patients on the health of each individual patient. One relevant measure would be involving former patients or starting up physical activity in groups. The findings show that the informal patient community among fellow patients is an important part of their social support system and thereby also may contribute to health and well-being. PMID- 17116150 TI - Post-stroke depression - effect on the life situation of the significant other. AB - The aim of this study was to describe the life situation among 'significant others' to patients with post-stroke depression, and to identify associations between the life situation of the significant others and patient characteristics. Seventy-one dyads consisting of patients with a diagnosed post-stroke depression and their significant others were included. The patients were assessed for depression with the Montgomery-Asberg Depression Rating Scale and diagnosed according to the DSM-IV. The assessments of significant others included their own life situation and proxy assessments of the patients' state of depression, anger, change of personality, and need of assistance. Significant others of male stroke patients reported a more negative impact on their life situation, than did significant others of female stroke patients (p = 0.04). There was a significant association between the patient's level of depression and physical function [activities of daily living (ADL)], with those with less impaired ADL having more major depression than those with more impaired ADL (p = 0.007). This study indicates that major post-stroke depression is more common among patients with limited functional deficits. This highlights the importance of assessment for depression also among seemingly recovered stroke patients in order to treat and support those in need. This study also stresses the importance of identifying different needs of the significant others in order to provide appropriate support in their caregiving role. PMID- 17116151 TI - 'Give me a break' - respite care for older carers of mentally ill persons. AB - BACKGROUND: The concept of respite care has been widely accepted and used throughout most developed countries, although primarily in connection with frail elderly, people with dementia, people with developmental disability or children. This paper aims at exploring older family carers' needs for, access to and use of respite services when caring for people with a mental illness (excluding dementia) in the community. METHOD: Data were collected through: semi-structured interviews with 26 mental health professionals, 21 family carers aged over 55 years and 25 respite service providers; demographic questionnaires; field notes; and local area, state and national documents relating to respite care services. Demographic data were analysed using descriptive statistics. All 72 interviews, field notes and other documents were analysed using content analysis. RESULTS: Current respite services are not adequate to meet the needs of mentally ill patients and older carers. The unique aspects of respite care for older carers of persons with mental illness are poorly understood. CONCLUSION: Bringing these issues to light creates the potential to address the unmet needs of older carers, improve provision of respite services, enhance quality of life of those who can benefit from respite services, and assist mental health professionals to work better with carers. PMID- 17116152 TI - Life satisfaction among informal caregivers in comparison with non-caregivers. AB - Being a caregiver with responsibility for someone with reduced health compared with not being a caregiver may mean different views of life satisfaction. Knowledge of what leads to reduced life satisfaction in caregivers may be helpful in interventions. Informal caregivers gainfully employed or not, aged 50-89 years, were studied with regard to life satisfaction depending on the extent of caregiving to identify types of social support of value for caregivers. A cross sectional study was conducted including a sample of 151 informal caregivers with a high caregiving extent, 392 caregivers with a lower caregiving extent and 1258 non-caregivers. The sample was randomly selected to correspond with older Swedes as a whole aged 50-89 years. A questionnaire containing a modified form of Older Americans' Resources Schedule (OARS) and Life Satisfaction Index Z (LSIZ) was used; 19.6% helped someone with activities of daily living and of these 27% stated that they did so frequently. Frequent caregiving implied significantly higher age, being more often married and retired, than less frequent caregivers and non-caregivers. Frequent caregivers also performed personal activities of daily living (PADL) to a higher extent than less frequent caregivers and had significantly lower LSIZ (mean 14.8) than less frequent caregivers (mean 17.6) and non-caregivers (mean 17.7). No significant differences were found between less frequent caregivers and non-caregivers in LSIZ. One-fourth had support from others, the commonest type being able to converse with a next of kin, and help and advice from professionals. Lower life satisfaction was associated with not being employed, low social resources, not refreshed after a night's sleep, overall poor health and frequent caregiving in the entire sample. High caregiving extent was associated with lower life satisfaction. The most important factors explaining lower life satisfaction among frequent caregivers were having low social resources and having poor health. Economic compensation or payment was the support most desired. PMID- 17116153 TI - School health nurses and substance use among adolescents - towards individual identification and early intervention. AB - Adolescents' health is today threatened by the use of alcohol and other psychoactive substances. It is therefore important to develop interventions related to substance use in school health care. The aim of this study was to examine the empowering or risk background factors related to substance use among adolescents, and the ability of school nurses (PHN) to identify these factors and to provide needed individual early intervention. The data were collected by semistructured questionnaires completed by 14- to 18-year-old adolescents (n = 326, response rate 79) and PHNs (n = 10) in 2004. The adolescent questionnaire consisted of items related to the respondents' background and Adolescents' Substance Use Measurement (ADSUME). Following individual consent, adolescents' ADSUME responses were sent to the PHNs for intervention. The PHNs assessed the adolescents' empowering background factors and intervention using the questionnaire, and 70% (n = 228) of their answers matched the adolescents' answers. The data were analysed with the SPSS software using the chi-squared test, Fisher's exact test, kappa coefficient and agreement percentages. Substance use among adolescents was associated with parental support, mother's education and smoking, the adolescents' knowledge about substances, peer support and hobbies. The PHNs' assessments regarding supportive background were not in agreement with the assessments of adolescents who were using hazardous substances. One-fifth of the adolescents received the brief intervention, although many of them might have needed extra support and follow-up on the basis of their ADSUME results. The research findings can be generalized only for alcohol use, because only 3% of the study informants used substances other than alcohol. Further research is warranted concerning PHNs' ability to identify hazardous substance use and to ensure preventive early intervention and requisite support among substance-using adolescents in order to improve evidence-based health promotion. PMID- 17116154 TI - Experiences of Swedish community health nurses working with health promotion and a patient-held health record. AB - Community health nurses have a tradition of preventive care, and might therefore be a key group in the introduction of new health-promotion methods. The aim of this study was to describe Swedish community health nurses' experiences in working with health promotion and a patient-held record as an integrated tool in their health-promotion work. Interviews were performed with 12 nurses at primary healthcare centres in the county of Dalarna, Sweden. A qualitative content analysis applying aspects of the grounded theory approach was performed. Central to the analysis was the nurses' struggle for balance, in being both a doer of practical disease-oriented tasks and a health-promotion communicator. Descriptions of the nurses' struggles to balance their work were grouped into three themes: (i) working alone and as a part of a team; (ii) nurse-related and patient-related interests; and (iii) patient's responsibility and shared responsibility between patient and nurse. The findings indicated that the structural organization in the primary healthcare centres was important for the community health nurses' means to work with health promotion and the patient-held record. In addition, the community health nurses' cognitive and emotional needs also affected this balance. In conclusion, the struggle of community health nurses to find a balance between being doers and health-promotion communicators is valuable in understanding health promotion in primary health care. The study indicates that it is not enough to develop health-promotion methods acceptable to community health nurses. A comprehensive examination of working conditions and the content of daily work is needed to ensure an emphasis on health promotion, including long-term usage of patient-held records. PMID- 17116155 TI - Properties of a Norwegian version of the Ward Organisational Features Scales. AB - To assess the association between nurses' practice environment and established measures of discharged patients' experiences during hospitalisation it is necessary to have a validated instrument to collect data on nurses' work environment in hospital wards. The purpose of this study was to assess the reliability and validity of a Norwegian version of the Ward Organisational Features Scales (WOFS), an instrument developed in the UK to describe environmental factors influential on the effectiveness of nursing services. After linguistic adaptation of the questionnaire, the reliability and validity were assessed in a sample of 99 nurses, of whom 46 (45%) responded. Most of the 12 tested scales showed acceptable internal consistency (10 had Cronbach's alpha > 0.70) and test-retest reliability (eight had intraclass correlation coefficient >0.70). The WOFS scales' correlations with external variables were in line with hypothesised correlations, thus supporting their validity. Scales describing nurses' influence on ward organisation and management did not perform well, probably due to differences in intraward organisation in the two countries. In spite of a limited sample size this study suggests that the majority of the scales of the WOFS showed satisfactory psychometric properties. They can be useful for assessing and describing aspects of nurses' work environment in Norwegian hospitals. PMID- 17116156 TI - Health in patients with Type 2 diabetes: an interview study based on the Welfare Theory of Health. AB - BACKGROUND: In his Welfare Theory of Health Nordenfelt describes health as a person's ability to fulfil vital goals in different life areas. In order to use the theory in nursing, a semi-structured interview guide was constructed including questions about which vital goals Type 2 diabetic patients have and believe are important for their own welfare in different life areas. AIM: The aim of the study was to carry out preliminary validation of the interview guide. Two hypotheses were formulated: (i) Dissatisfied or unhealthy diabetic persons score lower on health-related quality of life (HRQL) than do those classified as healthy; (ii) A person with diabetes who uses avoidance as his/her main coping strategy restructures his/her vital goals in order to avoid failure and thereby dissatisfaction increases. METHODS: Interviews with 155 randomly selected Type 2 diabetic patients were conducted using the interview guide. HRQL and coping were also measured. The interview data were analysed using content analysis. RESULTS: The results were in accordance with the health theory, i.e. a person with Type 2 diabetes may be unhealthy, but still satisfied with life as a whole. Our two hypotheses were confirmed: Dissatisfied or unhealthy subjects with Type 2 diabetes had HRQL scores in all domains but physical functioning and bodily pain that were lower than scores of those who were satisfied or healthy, and a person with Type 2 diabetes who uses avoidance as his/her main coping strategy does seem to restructure his/her vital goals to avoid failure. CONCLUSION: The theory-based interview guide tested in this study seemed to reflect the notion that health includes fulfilment of vital goals. PMID- 17116158 TI - Plasma levels of tumour necrosis factor-alpha in patients with chronic periodontitis and type 2 diabetes. AB - OBJECTIVES: Studies suggest that elevated circulating tumour necrosis factor alpha (TNF-alpha) may contribute to insulin resistance in patients with type 2 diabetes. The source of plasma TNF has been thought to be adipocytes associated with obesity, but inflammation and infection result in TNF-alpha production as well. METHODS: We studied 46 patients with type 2 diabetes and chronic periodontitis to determine the relationship between plasma TNF-alpha levels and clinical measures of periodontitis, gingival crevicular fluid (GCF) interleukin 1beta (IL-1beta), plasma endotoxin, serum glucose, and glycated haemoglobin (HbA1c). TNF-alpha levels were measured using a high sensitivity enzyme-linked immunosorbent assay. RESULTS: TNF-alpha showed a significant positive correlation with attachment loss (r=0.40, p=0.009), plasma endotoxin (r=0.33, p=0.03), and GCF IL-1beta (r=0.33, p=0.035), but not probing depth (r=0.28, p=0.07), bleeding on probing (r=0.30, p=0.053), plaque index (r=0.22, p=0.17), serum glucose, HbA1c (r=0.10, p=0.50), or body mass index (r=0.077, p=0.62). A dose-response relationship was observed between periodontitis severity and TNF-alpha (p=0.012). CONCLUSION: The finding that chronic periodontitis is associated with plasma TNF alpha levels in subjects with type 2 diabetes supports the hypothesis that periodontal infection and inflammation may contribute to insulin resistance. PMID- 17116159 TI - Meta-analyses of studies of 0.2% delmopinol mouth rinse as an adjunct to gingival health and plaque control measures. AB - BACKGROUND: Delmopinol is a third-generation anti-plaque agent used as a mouthwash to reduce plaque and alleviate gingivitis. OBJECTIVE: To create an overview of the anti-plaque efficacy of 0.2% delmopinol as an adjunct to normal oral hygiene measures by meta-analysis of completed clinical trials. MATERIALS AND METHODS: Eight double-blind, parallel-group studies were identified. Study durations ranged from 8 to 24 weeks. Five studies (n=913) involved supervised rinsing; three studies (n=467) involved unsupervised rinsing. These sets of trials were analysed separately and in combination. Efficacy outcomes comprised modified plaque index, modified gingival index (MGI) and gingival bleeding on probing (BOP). RESULTS: Delmopinol 0.2% was superior to placebo for the reduction of plaque scores in both sets of studies. Effects on MGI and BOP were also better with delmopinol 0.2% than with placebo. In most instances, 95% confidence intervals were wholly in favour of delmopinol. Pooled analysis of all eight studies confirmed statistically significant effects of delmopinol 0.2% compared with placebo (p<0.00001). Delmopinol met the efficacy criteria of the American Dental Association in studies of extended duration. CONCLUSION: Delmopinol 0.2% mouthwash is effective as an adjunct measure for reducing plaque burden and indices of gingivitis, whether or not it is used under supervision. PMID- 17116160 TI - Periodontal disease and perinatal outcomes: a case-control study. AB - AIM: Our aim was to measure the association of maternal periodontitis with low birth weight (LBW), pre-term LBW, and intra-uterine growth restriction. MATERIAL AND METHODS: An inclusive case-control design including subjects examined for periodontitis through attachment loss, information on perinatal outcomes and general health. Data were analysed through conditional logistic regression. RESULTS: Cases (n=304) and controls (n=611) had similar prevalence and severity of periodontitis, defined as at least three sites, in different teeth, with loss of three or more millimetres of clinical attachment level. Several factors were associated with the outcome, but the crude odds ratio for periodontitis was not significant. Odds ratio were 0.93 [95% confidence interval (CI): 0.63-1.41] for LBW and 0.92 (95% CI:0.54-1.57) for pre-term LBW in the presence of periodontitis, after adjustment for maternal age, previous pregnancies, pre-natal care, smoking, previous low birth or premature birth and other medical conditions, on a hierarchical model. CONCLUSIONS: Results do not support the hypothesis of association observed in previous studies after appropriate controlling for confounding variables. Negative peri-natal outcomes are better explained by determinants other than periodontal health. This study adds to the growing body of literature on the relationship between periodontal diseases and systemic health. PMID- 17116161 TI - Distribution of genotypes of Porphyromonas gingivalis in type 2 diabetic patients with periodontitis in Mexico. AB - OBJECTIVE: To determine and compare the distribution of Porphyromonas gingivalis fimA genotypes in type 2 diabetes mellitus (T2DM) patients affected by periodontitis, using non-diabetic subjects with and without periodontitis as control groups. MATERIAL AND METHODS: This study involved 75 subjects divided into three groups of 25 subjects each: Group 1 (non-T2DM without periodontitis), Group 2 (non-T2DM with periodontitis) and Group 3 (T2DM with periodontitis). The outcome variable was periodontitis, and explanatory variables were age, sex, T2DM and specific P. gingivalis fimA genotypes. RESULTS: In non-T2DM subjects with healthy periodontal tissues, type I fimA was the most frequently detected individually (40%) or in combinations (40%). In non-T2DM subjects with periodontitis, the most frequently detected type was Ib individually (20%) or in combinations (36%). In T2DM patients with periodontitis, the most frequently detected types were types I (20%) and III (20%), but there was no statistical difference (p>0.05) with non-T2DM periodontitis subjects. CONCLUSIONS: Type I genotype was more frequently detected in periodontally healthy sites from non T2DM subjects than in periodontitis sites from either subjects with or without T2DM. However, in sites affected by periodontitis from T2DM subjects the predominating types were I and III, which are less virulent strains of P. gingivalis. PMID- 17116162 TI - Isolation rearing induces recognition memory deficits accompanied by cytoskeletal alterations in rat hippocampus. AB - Social isolation from weaning affects hippocampal structure and function in the rat. The intrinsic dynamic instability of the cytoskeletal microtubular system is essential for neuronal development and organization. Accordingly, the present paper investigated the effects of social isolation on hippocampal levels of alpha tubulin isoforms associated with microtubule dynamics, the dendritic marker MAP-2 and alterations in locomotor activity and recognition memory. Male Lister Hooded rats (postnatal day 25-28) were housed either in groups or singly (isolated animals) for 30 days. Locomotor activity in a novel arena and novel object recognition were monitored in activity boxes. The hippocampus was dissected out 18 h after the novel object recognition task. Levels of alpha-tubulin isoforms and MAP-2 were analysed using Western blots. The experiments were conducted in duplicate, using two batches of rats obtained from different suppliers. Isolated animals were hyperactive and showed recognition memory deficits in the novel object recognition task. These behavioural alterations were accompanied by specific alterations in hippocampal alpha-tubulin isoforms and decreased MAP-2 expression. The results confirm that rearing rats in isolation produces hyperactivity and cognitive deficits. The behavioural alterations were accompanied by hippocampal cytoskeletal changes consistent with microtubule stabilization, and by decreased MAP-2 expression. These findings are indicative of an abnormal development of synaptic connections and/or reductions in neuronal cell number. The developmental structural abnormalities in the hippocampus may contribute to the cognitive impairments which result from isolation rearing in rats. PMID- 17116163 TI - Relationship between the perifornical hypothalamic area and oral pontine reticular nucleus in the rat. Possible implication of the hypocretinergic projection in the control of rapid eye movement sleep. AB - The perifornical (PeF) area in the posterior lateral hypothalamus has been implicated in several physiological functions including the regulation of sleep wakefulness. Some PeF neurons, which contain hypocretin, have been suggested to play an important role in sleep-wake regulation. The aim of the present study was to examine the effect of the PeF area and hypocretin on the electrophysiological activity of neurons of the oral pontine reticular nucleus (PnO), which is an important structure in the generation and maintenance of rapid eye movement sleep. PnO neurons were recorded in urethane-anesthetized rats. Extracellular recordings were performed by means of tungsten microelectrodes or barrel micropipettes. Electrical stimulation of the ipsilateral PeF area elicited orthodromic responses in both type I (49%) and type II (58%) electrophysiologically characterized PnO neurons, with a mean latency of 13.0 +/- 2 and 8.3 +/- 5 ms, respectively. In six cases, antidromic spikes were evoked in type I PnO neurons with a mean latency of 3.2 +/- 0.4 ms, indicating the existence of PnO neurons that projected to the PeF area. Anatomical studies showed retrogradely labeled neurons in the PeF area from the PnO. Some of these neurons projecting to the PnO contained hypocretin (17.8%). Iontophoretic application of hypocretin-1 through a barrel micropipette in the PnO induced an inhibition, which was blocked by a previous iontophoretic application of bicuculline, indicating that the inhibitory action of hypocretin-1 may be due to activation of GABA(A) receptors. These data suggest that the PeF area may control the generation of rapid eye movement sleep through a hypocretinergic projection by inhibiting the activity of PnO neurons. PMID- 17116164 TI - Phonological but not auditory discrimination is impaired in dyslexia. AB - Deficient phonological skills are considered to be a core problem in developmental dyslexia. Children with dyslexia often demonstrate poorer performance than non-impaired readers when categorizing speech-sounds. Using the automatic mismatch response, we show that in contrast to this deficit at the behavioural level, neurophysiological responding in dyslexic children indicates their ability to automatically discriminate syllables. Therefore, the phonological deficit is unlikely to be caused by a temporal deficit or by a noisy functional organization in the respective representational cortex. We obtained measures of reading, spelling and categorical speech-perception from 58 dyslexic children and 21 control children. The children also participated in magnetoencephalographic measurements while being stimulated acoustically with the syllables /ba/ and /da/ in an oddball paradigm. Mismatch field (MMF) amplitudes between standard and deviant stimuli were obtained. Dyslexic children performed more poorly than control children on all test measures. However, the groups did not differ in MMF amplitude or latency. No correlations were found between MMF amplitudes and behavioural performance. These results were obtained with a large sample size and thus speak robustly against a general deficit in auditory discrimination in dyslexia. These results are compatible with the idea that decoding difficulties occur later in the processing stream where access to the phonological lexicon is attempted. PMID- 17116165 TI - Bidirectional regulation of dopamine D2 and neurotensin NTS1 receptors in dopamine neurons. AB - Several lines of evidence suggest a close association between dopamine (DA) and neurotensin (NT) systems in the CNS. Indeed, in the rodent brain, abundant NT containing fibres are found in DA-rich areas such as the ventral tegmental area and substantia nigra. Moreover, it has been shown in vivo that NT, acting through its high-affinity receptor (NTS1), reduces the physiological and behavioural effects of DA D2 receptor (D2R) activation, a critical autoreceptor feedback system regulating DA neurotransmission. However, the mechanism of this interaction is still elusive. The aim of our study was thus to reproduce in vitro the interaction between D2R and NTS1, and then to characterize the mechanisms implicated. We used a primary culture model of DA neurons prepared from transgenic mice expressing green fluorescent protein under the control of the tyrosine hydroxylase promoter. In these cultures, DA neurons endogenously express both D2R and NTS1. Using electrophysiological recordings, we show that activation of D2R directly inhibits the firing rate of DA neurons. In addition, we find that NT, acting through a NTS1-like receptor, is able to reduce D2R autoreceptor function independently of its ability to enhance DA neuron firing, and that this interaction occurs through a protein kinase C- and Ca(2+)-dependent mechanism. Furthermore, prior activation of D2R reduces the ability of NTS1 to induce intracellular Ca(2+) mobilization. Our findings provide evidence for bidirectional interaction between D2R and NTS1 in DA neurons, a regulatory mechanism that could play a key role in the control of the activity of these neurons. PMID- 17116166 TI - The effects of stubble retention and nitrogen application on soil microbial community structure and functional gene abundance under irrigated maize. AB - The effects of agronomic management practices on the soil microbial community were investigated in a maize production system in New South Wales, Australia. The site has been intensively studied to measure the impact of stubble management and N-fertilizer application on greenhouse gas emissions (CO(2) and N(2)O), N cycling, pathology, soil structure and yield. As all of these endpoints can be regulated by microbial processes, the microbiology of the system was examined. Soil samples were taken after a winter fallow period and the diversity of the bacterial and fungal communities was measured using PCR-denaturing gradient gel electrophoresis. Stubble and N shifted the structure of bacterial and fungal communities with the primary driver being stubble addition on the fungal community structure (P<0.05 for all effects). Changes in C, N (total and NO(3)), K and Na, were correlated (P<0.05) with variation in the microbial community structure. Quantitative PCR showed that nifH (nitrogen fixation) and napA (denitrification) gene abundance increased upon stubble retention, whereas amoA gene numbers were increased by N addition. These results showed that the management of both stubble and N have significant and long-term impacts on the size and structure of the soil microbial community at phylogenetic and functional levels. PMID- 17116168 TI - Strain specificity and cholinergic modulation of visuospatial attention in three inbred mouse strains. AB - The tremendous increase in the use of mouse inbred strains and mutant mice to study the molecular basis of psychiatric disorders urges for a better understanding of attentional performance in mice. To this aim, we investigated possible strain differences in performance and cholinergic modulation of visuospatial attention in three widely used mouse inbred strains (129S2/SvHsd, C57BL/6JOlaHsd and DBA/2OlaHsd) in the five-choice serial reaction time task. Results indicated that after extended training, performance of 129S2/SvHsd mice was superior to that of C57BL/6JOlaHsd and DBA/2OlaHsd mice in terms of attention, omission errors, inhibitory control and latencies to correct choice. Increasing the attentional load resulted in comparable decrements in attention in all strains and inhibitory control impairments that were most pronounced in DBA/2OlaHsd mice. Further pharmacological evaluation indicated that all strains showed attentional impairments after treatment with the muscarinic and nicotinic antagonists scopolamine and mecamylamine, respectively. 129S2/SvHsd mice were less sensitive, whereas DBA/2OlaHsd mice appeared more sensitive to the detrimental effects of mecamylamine. In addition, subchronic, but not acute, nicotine treatment slightly improved attentional performance in all strains to the same extent. In conclusion, our data indicate strain specificity with particularly good performance of 129S2/SvHsd mice and strong cholinergic involvement in visuospatial attention in mice. PMID- 17116169 TI - Impaired attentional modulation of auditory evoked potentials in N-methyl-D aspartate NR1 hypomorphic mice. AB - In human neurophysiology, auditory event-related potentials (AEPs) are used to investigate cognitive processes such as selective attention. Selective attention to specific tones causes a negative enhancement of AEPs known as processing negativity (PN), which is reduced in patients with schizophrenia. The evidence suggests that impaired selective attention in these patients may partially depend on deficient N-methyl-D-aspartate receptor (NMDAR)-mediated signaling. The goal of this study was to corroborate the involvement of the NMDAR in selective attention using a mouse model. To this end, we first investigated the presence of PN-like activity in C57BL/6J mice by recording AEPs during a fear-conditioning paradigm. Two alternating trains of tones, differing in stimulus duration, were presented on 7 subsequent days. One group received a mild foot shock delivered within the presentation of one train (conditioning train) on days 3-5 (conditioning days), while controls were never shocked. The fear-conditioned group (n= 9) indeed showed a PN-like activity during conditioning days manifested as a significant positive enhancement in the AEPs to the stimuli in the conditioning train that was not observed in the controls. The same paradigm was then applied to mice with reduced expression of the NMDAR1 (NR1) subunit and to a wild-type control group (each group n= 6). The NR1 mutants showed an associative AEP enhancement, but its magnitude was significantly reduced as compared with the magnitude in wild-type mice. We conclude that electrophysiological manifestations of selective attention are observable yet of different polarity in mice and that they require intact NMDAR-mediated signaling. Thus, deficient NMDAR functioning may contribute to abnormal selective attention in schizophrenia. PMID- 17116170 TI - MEKK3 is essential for lipopolysaccharide-induced interleukin-6 and granulocyte macrophage colony-stimulating factor production in macrophages. AB - Mitogen-activated protein/ERK kinase kinase 3 (MEKK3) is a Ser/Thr protein kinase belonging to the MEKK/STE11 subgroup of the MAP3K family. Recently, we found that MEKK3 plays a critical role in interleukin-1 (IL-1) receptor and Toll-like receptor 4 signalling using established primary mouse embryonic fibroblast (MEF) cell lines. However, the function of MEKK3 in immune cells has not been studied because germ-line MEKK3 knockout mice are embryonically lethal between embryonic days 10 and 11. In this study, we used small interference RNA to the mouse Mekk3 gene to specifically knock down MEKK3 expression in the macrophage line Raw264.7. We found that the lipopolysaccharide-induced IL-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF) production was dramatically decreased in MEKK3 knockdown cells whereas the tumour necrosis factor-alpha and IL-1beta production were not affected. We also observed that the ERK1/2, p38 and JNK MAPK induction in MEKK3 knockdown cells were moderately inhibited within the first 60 min of stimulation, while the ERK and p38 were more severely inhibited after 2-4 hr of stimulation. Degradation of IkappaBalpha was also partially blocked in MEKK3 knockdown cells. Notably, the impairment in IL-6 and GM-CSF production in the MEKK3 knockdown cells was restored by reintroducing a human Mekk3 cDNA that could not be targeted by mouse Mekk3-siRNAs. In conclusion, this study showed that MEKK3 is a crucial and specific regulator of the proinflammatory cytokines IL-6 and GM-CSF in macrophages and provided a novel method for investigating MEKK3 function in other immune cells. PMID- 17116171 TI - Increased proteolysis of diphtheria toxin by human monocytes after heat shock: a subsidiary role for heat-shock protein 70 in antigen processing. AB - The expression of heat-shock proteins (hsp) increases after exposure to various stresses including elevated temperatures, oxidative injury, infection and inflammation. As molecular chaperones, hsp have been shown to participate in antigen processing and presentation, in part through increasing the stability and expression of major histocompatibility complex molecules. Heat shock selectively increases human T-cell responses to processed antigen, but does not affect T-cell proliferation induced by non-processed antigens. Here, we have analysed the mechanisms by which stress such as heat shock, and the ensuing hsp over expression affect the processing of diphtheria toxin (DT) in peripheral blood monocytes. We found that heat shock increased DT proteolysis in endosomes and lysosomes while the activities of the cathepsins B and D, classically involved in DT proteolysis, were decreased. These effects correlated with the heat-shock mediated increase in hsp 70 expression observed in endosomes and lysosomes. Actinomycin D or blocking anti-hsp 70 antibodies abolished the heat-shock mediated increase in DT proteolysis. These data indicate that the increased expression of hsp 70 constitutes a subsidiary mechanism that facilitates antigen proteolysis in stressed cells. Confirming these data, presentation by formaldehyde-fixed cells of DT proteolysates that were obtained with endosomes and lysosomes from heat-shocked peripheral blood monocytes showed higher stimulation of T cells than those generated with endosomes and lysosomes from control peripheral blood monocytes. PMID- 17116172 TI - Differential type I interferon activation and susceptibility of dendritic cell populations to porcine arterivirus. AB - Dendritic cells (DCs) play a role in anti-viral immunity by providing early innate protection against viral replication and by presenting antigen to T cells for initiation of the adaptive immune response. Studies show the adaptive response to porcine reproductive and respiratory syndrome virus (PRRSV) is ineffective for complete viral elimination. Other studies describe the kinetics of the adaptive response to PRRSV, but have not investigated the early response by DCs. We hypothesize that there is an aberrant activation of DCs early in PRRSV infection; consequently, the adaptive response is triggered inadequately. The current study characterized a subtype of porcine lung DCs (L-DCs) and investigated the ability of PRRSV to infect and replicate in L-DCs and monocyte derived DCs (MDDCs). Furthermore, the type I interferon anti-viral response to PRRSV with and without the addition of recombinant porcine IFN-alpha (rpIFN alpha), an important cytokine that signals for anti-viral mediator activation, was analysed. Results show that PRRSV replicated in MDDCs but not L-DCs, providing evidence that these cells have followed distinct differentiation pathways. Although both cell types responded to PRRSV with an induction of IFN beta mRNA, the magnitude and duration of the response differed between cell types. The addition of rpIFN-alpha was protective in MDDCs, and mRNA synthesis of Mx (myxovirus resistant) and PKR (double-stranded RNA dependent protein kinase) was observed in both cell types after rpIFN-alpha addition. Overall, PRRSV replicated in MDDCs but not L-DCs, and rpIFN-alpha was required for the transcription of protective anti-viral mediators. DC response to PRRSV was limited to IFN-beta transcription, which may be inadequate in triggering the adaptive immune response. PMID- 17116173 TI - Suppression of major histocompatibility complex class II-associated invariant chain enhances the potency of an HIV gp120 DNA vaccine. AB - Summary One function of the major histocompatibility complex (MHC) class II associated invariant chain (Ii) is to prevent MHC class II molecules from binding endogenously generated antigenic epitopes. Ii inhibition leads to MHC class II presentation of endogenous antigens by APC without interrupting MHC class I presentation. We present data that in vivo immunization of BALB/c mice with HIV gp120 cDNA plus an Ii suppressive construct significantly enhances the activation of both gp120-specific T helper (Th) cells and cytotoxic T lymphocytes (CTL). Our results support the concept that MHC class II-positive/Ii-negative (class II(+)/Ii(-)) antigen-presenting cells (APC) present endogenously synthesized vaccine antigens simultaneously by MHC class II and class I molecules, activating both CD4(+) and CD8(+) T cells. Activated CD4(+) T cells locally strengthen the response of CD8(+) CTL, thus enhancing the potency of a DNA vaccine. PMID- 17116174 TI - Differential polarization of immune responses by genetic cotransfer of chemokines changes the protective immunity of DNA vaccine against pseudorabies virus. AB - Chemokines play a key role in eliciting adaptive immune responses by selectively attracting the innate cellular components to the site of antigen presentation. To evaluate the effect of the genetic adjuvant of chemokines on the adaptive immune responses induced by a plasmid DNA vaccine expressing glycorotein B (gB) of the pseudorabies virus (PrV), a PrV DNA vaccine was co-inoculated with plasmid DNA expressing certain chemokines including CCL3 (MIP-1alpha), CCL4 (MIP-1beta), CCL5 (RANTES), CXCL8 (MIP-2), and CXCL10 (IP-10). A co-injection of the CCL3 plasmid DNA induced immunity that was biased to the T helper type 2 (Th2) pattern, as judged by the ratio of immunoglobulin G isotypes and the production of interleukin-4 cytokine generated from stimulated immune T cells. However, CCL5 and CXCL10 induced immune responses of the Th1-type, which rendered the recipients more resistant to a virulent virus infection. CXCL8 also showed enhanced humoral and cell-mediated immunity (mixed-type pattern) providing effective protection against a viral challenge. However, there was no change in the immune responses induced by the PrV DNA vaccine in CCL4 recipients. These results suggest that co-injection of a chemokine, in the form of an adjuvant preparation, causes a rebalancing of the immunity, which subsequently affects the protective efficacy against a virulent virus infection. PMID- 17116175 TI - Navigating services for people with diabetes through the storms of health-care policy development. PMID- 17116176 TI - Cortisol--cause and cure for metabolic syndrome? AB - Similarities between the metabolic syndrome and Cushing's syndrome, and reversibility of the features of Cushing's syndrome, suggest that cortisol may contribute to the pathophysiology of both conditions and that reducing cortisol action may provide a novel therapeutic approach in the metabolic syndrome. There is substantial evidence that circulating cortisol concentrations are higher in people with hypertension and glucose intolerance. The basis for this activation of the hypothalamic-pituitary-adrenal axis remains uncertain, but it may be attributable to 'programming' effects of events in early life, since it is associated with low birth weight. In obese people, intracellular cortisol levels within adipose tissue are further amplified by increased local regeneration of cortisol by the enzyme 11beta-HSD type 1. In mice, transgenic manipulations of 11beta-HSD1 have potent effects on obesity and associated features of the metabolic syndrome. Promising preclinical data suggest that novel 11beta-HSD1 inhibitors will have a role in lowering intracellular cortisol levels as a treatment for the metabolic syndrome. In addition to their metabolic effects, glucocorticoids act in the blood vessel wall. Pharmacoepidemiological studies suggest that glucocorticoid excess is an independent risk factor for cardiovascular disease. Recent data suggest that 11beta-HSD1 within the blood vessel wall influences vascular remodelling and angiogenesis, for example in the myocardium following coronary artery occlusion. Thus, glucocorticoid signalling provides a potentially tractable system to influence both risk factors for, and the outcome of, Type 2 diabetes and cardiovascular disease. PMID- 17116177 TI - A Diabetes Outcome Progression Trial (ADOPT): baseline characteristics of Type 2 diabetic patients in North America and Europe. AB - AIMS: To examine baseline characteristics of patients recruited into ADOPT, a multinational trial comparing three oral glucose-lowering monotherapies. METHODS: Between April 2000 and June 2002, 4360 patients aged 30-75 years with Type 2 diabetes diagnosed for < 3 years and remaining on diet therapy alone with fasting plasma glucose levels (FPG) between 7.0 and 10.0 mmol/l were enrolled by 488 North American and European centres. Medical histories, anthropometric data and laboratory measurements were determined using common methodologies. RESULTS: The mean (SD) age of the patients was 57 (10) years, body mass index 32.2 (6.4) kg/m(2), HbA(1c) 7.4 (0.9)%; 58% were male, 88% Caucasian and 15% smoked. North American Caucasians (NAC) were younger, more obese, and more insulin resistant than European Caucasians (EUC), but had better pancreatic B-cell function. NAC had lower total, low-density lipoprotein- and high-density liporpotein cholesterol concentrations with higher triglyceride concentrations and were more often on lipid-lowering treatment. They had lower blood pressure levels but were equally likely to be on antihypertensive treatment. Metabolic syndrome was more frequent and microalbuminuria less frequent in NAC. Within North America, NAC had lower HbA(1c) concentrations than Blacks, Hispanics and Asians despite similar or higher FPG and 30-min postchallenge glucose concentrations. CONCLUSIONS: Caucasian North American and European ADOPT patients differ with respect to adiposity, insulin resistance and metabolic syndrome prevalence. North American Blacks, Hispanics and Asians had lower HbA(1c) concentrations than NAC despite similar or higher glucose concentrations. These phenotypic differences may influence the progression of Type 2 diabetes and the response to initial oral glucose-lowering monotherapy. PMID- 17116178 TI - HNF-1alpha G574S is a functional variant with decreased transactivation activity. AB - AIM: To assess the functional consequence of the hepatocyte nuclear factor 1alpha gene (HNF-1alpha) G574S variant previously proposed as a diabetes susceptibility allele, in a group of Mexican Type 2 diabetic patients with end-stage renal disease (ESRD). METHODS: The transcriptional activity of the HNF-1alpha G574S recombinant protein on the human insulin promoter was assessed by transfection assays in RINm5f and HepG2 cell lines. RESULTS: Two unrelated Mexican diabetic patients with no known African ancestry were found to carry the G574S variant. This substitution was not found among unrelated healthy control subjects. Whereas the G574S HNF-1alpha transcription activation of the human insulin promoter was 40% lower than that of the wild-type protein in RINm5f beta cells, no difference was found in a hepatic cell line (HepG2). CONCLUSIONS: G574S affects the transactivation potential of HNF-1alpha on the insulin promoter in pancreatic beta-cells. Although it has been difficult to prove its role in the development of diabetes in case-control association studies, this variant exhibits functional effects consistent with it being a potential diabetes susceptibility allele. PMID- 17116179 TI - Hepatocyte nuclear factor-1 beta mutations cause neonatal diabetes and intrauterine growth retardation: support for a critical role of HNF-1beta in human pancreatic development. AB - AIM: The transcription factor hepatocyte nuclear factor-1beta (HNF-1beta) is expressed in rodent pancreatic progenitor cells, where it is an important member of the genetic hierarchy that regulates the generation of pancreatic endocrine and exocrine cells. The recent description of an HNF-1beta mutation in a patient with neonatal diabetes suggests that HNF-1beta may also play a key role in human pancreatic B-cell development. We aimed to investigate the role of HNF-1beta mutations in neonatal diabetes and also the impact of HNF-1beta mutations on fetal growth. METHODS: We sequenced the HNF-1beta gene in 27 patients with neonatal diabetes in whom other known genetic aetiologies had been excluded. Birth weight was investigated in 21 patients with HNF-1beta mutations. RESULTS: A heterozygous HNF-1beta mutation, S148L, was identified in one patient with neonatal diabetes diagnosed at 17 days, which rapidly resolved only to relapse at 8 years. This patient had pancreatic atrophy, mild exocrine insufficiency and low birth weight (1.83 kg at 40 weeks' gestation). Intrauterine growth was markedly reduced in patients born to unaffected mothers with a median birth weight of 2.4 kg (range 1.8-3.3) (P = 0.006), median centile weight 3 (0.008-38), and 69% were small for gestational age. CONCLUSION: HNF-1beta mutations are a rare cause of neonatal diabetes as well as pancreatic exocrine and endocrine dysfunction. Low birth weight is a common feature of patients with HNF-1beta mutations and is consistent with reduced insulin secretion in utero. These findings support a key role for HNF-1beta in early pancreatic progenitor cells in man. PMID- 17116180 TI - Peroxisome proliferator-activated receptor (PPAR) delta genetic polymorphism and its association with insulin resistance index and fasting plasma glucose concentrations in Chinese subjects. AB - AIMS: Previous studies have shown that the peroxisome proliferator-activated receptor delta (PPARD) genetic polymorphism affects cholesterol metabolism in Whites. This association was not observed in a Korean population in a separate study, but this study showed a link between the PPARD polymorphism and body weight and fasting plasma glucose. The purpose of this study was to determine whether polymorphisms of PPARD influence glucose and cholesterol metabolism in Chinese subjects. We investigated the association between the polymorphism ( 87T/C) of the human PPARD gene and phenotypes related to body weight, insulin sensitivity, glucose and lipid metabolism in Chinese subjects. METHODS: Unrelated Chinese subjects (n = 663) in Shanghai were studied; 287 had newly diagnosed Type 2 diabetes mellitus and 376 were non-diabetic control subjects over 40 years old. Clinical parameters were collected and genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: In normal glucose tolerant (NGT) subjects, the C allele carriers had higher fasting plasma glucose concentrations (P = 0.0078) and a lower insulin sensitivity index (ISI) (P = 0.0365). The C allele carriers also showed higher concentrations of low density lipoprotein cholesterol (P = 0.0261) and percentage of body fat (P = 0.0357). There was a trend towards higher visceral adiposity in C allele carriers, but the difference was not significant (P = 0.0830). In diabetes patients, similar results were detected for plasma glucose concentrations (fasting plasma glucose P < 0.0001, 2-h plasma glucose P = 0.0052) and insulin sensitivity (homeostasis model assessment of insulin resistance P = 0.0094; ISI P = 0.0058). CONCLUSION: The PPARD-87T/C polymorphism is associated with higher fasting plasma glucose concentrations in both NGT and diabetic subjects, largely due to impaired insulin sensitivity. PMID- 17116181 TI - Mortality in individuals with Type 2 diabetes and heart disease in a unique New Zealand population. AB - AIMS: To determine rate and predictors of mortality in patients with Type 2 diabetes living in a population with a large proportion of Maori and Pacific peoples who were admitted to hospital with myocardial infarction (MI) or congestive cardiac failure (CCF). METHODS: The study population included 4193 individuals with Type 2 diabetes who lived in South Auckland and who participated in a primary care audit between 1994 and 1999. We studied a subgroup of 319 patients who subsequently had a hospital admission for MI or CCF between 1999 and 2001. We examined their demographics, drug treatment, vascular risk factors and mortality up to 2003. RESULTS: Following discharge, the prescription of beta blockers, angiotensin-converting enzyme (ACE) inhibitors, aspirin and statins had all increased significantly. Vascular risk factors including total cholesterol, high-density lipoprotein (HDL) cholesterol, systolic blood pressure and glycated haemoglobin (HbA(1c)) improved significantly. Nevertheless, mortality remained high with individuals admitted because of an MI 7.2-fold more likely to die early and those with a CCF admission 5.9-fold more likely to die early than other individuals with Type 2 diabetes. Maori patients have an 80% excess mortality. CONCLUSIONS: Patients with past admission for MI and/or CCF remain at exceptionally high risk of death for at least 4 years after hospital admission, even with improvements in management of cardiovascular risk factors. Maori individuals are at particular risk. PMID- 17116182 TI - Gestational diabetes: utility of fasting plasma glucose as a screening test depends on the diagnostic criteria. AB - AIMS: To demonstrate the effect of diagnostic criteria, as defined by four international expert panels, on the usefulness of fasting plasma glucose (FPG) as a screening test for gestational diabetes mellitus (GDM). METHODS: We tested 4602 pregnant women using a 75-g oral glucose tolerance test (OGTT) for universal GDM screening. The area under the receiver operating characteristic curve (AUC) was used to determine the FPG performance to detect GDM by the criteria of the American Diabetes Association (ADA), the Australasian Diabetes in Pregnancy Society, the European Association for the Study of Diabetes, and the World Health Organization (WHO). RESULTS: By applying ADA, Australasian, European and WHO criteria, respectively, the FPG: (i) AUC (95% CI) was 0.882 (0.866-0.897), 0.830 (0.809-0.852), 0.808 (0.791-0.825) and 0.690 (0.670-0.710); (ii) independently could 'rule-in' GDM (with 100% specificity) in 74 (10.9%), 620 (53.5%), 252 (45.3%) and 74 (7.6%) women; (iii) independently could 'rule-out' GDM in an additional 2864 (62.2%), 928 (20.2%), 1510 (32.8%) and 1171 (25.4%) women, at FPG thresholds (with 85% sensitivity); (iv) false-positive rate (FPR) was 29.4, 75.5, 63.8 and 71.2%, at these thresholds. CONCLUSIONS: The value of the FPG as a screening test for GDM is highly dependent on the diagnostic criteria. The performance is excellent with the ADA criteria. With the other criteria, the high FPR (poor specificity) would limit its utility as a screening test. Regardless of the criteria used, initial testing by FPG can significantly decrease the number of cumbersome OGTTs needed for the diagnosis of GDM. PMID- 17116183 TI - Reduced insulin secretion in normoglycaemic patients with beta-thalassaemia major. AB - AIMS: To assess insulin sensitivity and secretion in the fasting state in regularly transfused patients with beta-thalassaemia major with normal glucose response during an oral glucose tolerance test and to estimate its possible relation to iron overload. METHODS: We measured fasting glucose, insulin and C peptide levels in 24 patients with beta-thalassaemia major and 18 control subjects matched for age and body mass index. Insulin sensitivity and insulin release index were calculated according to the homeostasis model assessment (HOMA). Correlations with age, body mass index and serum ferritin were also calculated. RESULTS: Fasting glucose levels in patients were increased compared with control subjects (5.5 +/- 0.12 vs. 4.7 +/- 0.13 mmol/l, mean +/- SEM, P < 0.001). Pancreatic B-cell insulin secretion in the fasting state (estimated by SC(HOMA)) was lower in thalassaemic patients (SC(HOMA) 88.5 +/- 11.11 vs. 184.3 +/- 23.72 in control subjects, P < 0.001). Patients were then divided into those with impaired (IFG) and normal (NFG) fasting glucose. SC(HOMA) was higher in the patients with NFG compared with those with IFG patients (110.6 +/- 17.63 vs. 66.3 +/- 10.88, respectively, P < 0.05) but estimated insulin sensitivity (ISI(HOMA)) was similar. Plasma values of C-peptide correlated positively with ferritin (r = 0.42, P = 0.04) and SC(HOMA) (r = 0.45, P = 0.02) and negatively with ISI(HOMA) (r = -0.43, P = 0.03). CONCLUSIONS: These results support the concept that impaired B-cell function, as reflected by a reduction in the insulin secretion index, is present in beta-thalassaemic patients with normoglycaemia before changes in oral glucose tolerance tests are apparent. PMID- 17116184 TI - A randomized controlled trial of Sweet Talk, a text-messaging system to support young people with diabetes. AB - AIMS: To assess Sweet Talk, a text-messaging support system designed to enhance self-efficacy, facilitate uptake of intensive insulin therapy and improve glycaemic control in paediatric patients with Type 1 diabetes. METHODS: One hundred and twenty-six patients fulfilled the eligibility criteria; Type 1 diabetes for > 1 year, on conventional insulin therapy, aged 8-18 years. Ninety two patients were randomized to conventional insulin therapy (n = 28), conventional therapy and Sweet Talk (n = 33) or intensive insulin therapy and Sweet Talk (n = 31). Goal-setting at clinic visits was reinforced by daily text messages from the Sweet Talk software system, containing personalized goal specific prompts and messages tailored to patients' age, sex and insulin regimen. RESULTS: HbA(1c) did not change in patients on conventional therapy without or with Sweet Talk (10.3 +/- 1.7 vs. 10.1 +/- 1.7%), but improved in patients randomized to intensive therapy and Sweet Talk (9.2 +/- 2.2%, 95% CI -1.9, -0.5, P < 0.001). Sweet Talk was associated with improvement in diabetes self-efficacy (conventional therapy 56.0 +/- 13.7, conventional therapy plus Sweet Talk 62.1 +/ 6.6, 95% CI +2.6, +7.5, P = 0.003) and self-reported adherence (conventional therapy 70.4 +/- 20.0, conventional therapy plus Sweet Talk 77.2 +/- 16.1, 95% CI +0.4, +17.4, P = 0.042). When surveyed, 82% of patients felt that Sweet Talk had improved their diabetes self-management and 90% wanted to continue receiving messages. CONCLUSIONS: Sweet Talk was associated with improved self-efficacy and adherence; engaging a classically difficult to reach group of young people. While Sweet Talk alone did not improve glycaemic control, it may have had a role in supporting the introduction of intensive insulin therapy. Scheduled, tailored text messaging offers an innovative means of supporting adolescents with diabetes and could be adapted for other health-care settings and chronic diseases. PMID- 17116185 TI - Lipo-prostaglandin E1 in combination with steroid therapy is effective for treatment of sudden sensorineural hearing loss in Korean patients with Type 2 diabetes. AB - BACKGROUND: Diabetes mellitus is a risk factor for sudden sensorineural hearing loss (SSNHL), possibly as a result of microvascular damage, increased blood viscosity and embolic and thrombotic episodes. Aim To perform a prospective, randomized clinical trial to determine the therapeutic effect of lipo prostaglandin E(1) (lipo-PGE(1)) on SSNHL in Korean patients with Type 2 diabetes. METHODS: The study group consisted of 270 consecutive Korean patients (130 male, 140 female) in whom SSNHL was diagnosed. Patients were classified as diabetic or non-diabetic by the new American Diabetes Association criteria. With the approval of the institute ethics committee, patients were randomly assigned to treatment with of 10 microg lipo-PGE(1) (lipo-PGE(1) group) given as a continuous infusion over 5 days, or saline (placebo group). In addition, all patients studied were treated with 48 mg methylprednisolone for 5 days. RESULTS: The overall recovery rates after treatment were 51.5% in diabetic and 64.2% in non-diabetic patients. In diabetic patients, there was a significantly higher rate of hearing improvement in the lipo-PGE(1) group (64.7%) compared with the placebo group (37.5%), whereas, in non-diabetic patients, there was no significant difference in rate of hearing improvement between the lipo-PGE(1) group (70.3%) and the placebo group (58.3%). CONCLUSION: Lipo-PGE(1) may have beneficial effects in the treatment of SSNHL in Korean patients with Type 2 diabetes. PMID- 17116186 TI - Oxidative burden in prediabetic and diabetic individuals: evidence from plasma coenzyme Q(10). AB - AIM: Individuals with diabetes and prediabetes are at risk of vascular injury. However, the exact mechanisms are unclear. The mitochondria mobile electron carrier coenzyme Q(10) (CoQ(10)) is a potent lipophilic antioxidant. We hypothesize that oxidative stress, detectable as changes in plasma CoQ(10) concentrations and composition, plays an important role in vascular disease in diabetes. METHODS: We measured plasma CoQ(10) concentrations (including reduced ubiquinol and oxidized ubiquinone subfractions) in 60 subjects with normal glucose tolerance [NGT; fasting plasma glucose (FPG) < 5.5 mmol/l], 63 with impaired fasting glucose (IFG; FPG 5.6-6.9 mmol/l) and 69 with Type 2 diabetes (DM; FPG > 6.9 mmol/l). RESULTS: In men and women, the total CoQ(10)/total cholesterol ratio was reduced in DM (mean +/-sd) [male (M) 0.09 +/- 0.04; female (F) 0.07 +/- 0.04] compared with NGT (0.29 +/- 0.08; 0.21 +/- 0.07) and IFG (0.27 +/- 0.07; 0.23 +/- 0.07) (DM vs. NGT and IFG P = 0.001). A stepwise reduction in the plasma ubiquinol fraction (ubiquinol/total CoQ10) was observed from NGT (M 0.93 +/- 0.06; F 0.95 +/- 0.06) compared with IFG (0.43 +/- 0.25; 0.41 +/- 0.15) and DM (0.24 +/- 0.11; F 0.29 +/- 0.16) (DM vs. IFG vs. NGT P = 0.001). In contrast, the plasma ubiquinone/ubiquinol ratio increased from NGT (M 0.08 +/- 0.07, F 0.06 +/- 0.08) to IFG (2.14 +/- 1.84, 1.75 +/- 1.04) to DM (4.77 +/- 4.88, 3.81 +/- 3.71) (DM vs. IFG vs. NGT P = 0.001). These differences remained after adjusting for age, body mass index and FPG. CONCLUSIONS: The change in CoQ(10) with increasing FPG concentration suggests an increase in oxidative burden, already evident in the prediabetic IFG individuals. This increase in oxidative stress might contribute to the increased risk of vascular disease. PMID- 17116187 TI - Plasma homocysteine, oxidative stress and endothelial function in patients with Type 1 diabetes mellitus and microalbuminuria. AB - AIMS: The purpose of this study was to examine the associations between endothelial function, plasma homocysteine and oxidative stress in patients with Type 1 diabetes mellitus (DM) and microalbuminuria compared with DM patients with normoalbuminuria and non-diabetic control subjects. We wished to test the hypothesis that increased cardiovascular risk in patients with Type 1 diabetes and microalbuminuria may be in part as a result of hyperhomocysteinaemia-mediated oxidative stress leading to impaired endothelial function. METHODS: We measured forearm blood flow, total plasma homocysteine, total antioxidant status (TAOS) and whole blood glutathione in 31 DM patients, 16 with microalbuminuria and 15 with normoalbuminuria, and 15 non-diabetic control subjects. RESULTS: Plasma homocysteine levels were significantly higher in the microalbuminuric diabetic patients compared with the normoalbuminuric patients and the control subjects. TAOS was significantly lower in the micoalbuminuric and normoalbuminuric diabetic patients compared with the control subjects, although TAOS levels were similar in both groups of diabetic patients. There was no difference in forearm blood flow between the groups and no association between measured endothelial function and antioxidant defence/oxidative stress and homocysteine in each group. There was no association between plasma total homocysteine and TAOS or whole blood glutathione within the groups. CONCLUSIONS: We have found mild hyperhomocysteinaemia in microalbuminuric DM patients compared with normoalbuminuric DM patients and non diabetic subjects and some evidence for reduced antioxidant defence in DM patients. These findings add to our understanding of the increased risk of vascular disease in patients with Type 1 diabetes. PMID- 17116188 TI - Haemoglobin A1c is not a surrogate for glucose and insulin measures for investigating the early life and childhood determinants of insulin resistance and Type 2 diabetes in healthy children. An analysis from the Avon Longitudinal Study of Parents and Children (ALSPAC). AB - AIMS: Research into early life and childhood determinants of insulin resistance and Type 2 diabetes are complicated by requirements for fasting blood samples and glucose tolerance tests. We investigated haemoglobin A(1c) (HbA(1c)), a marker of glycaemia measured in non-fasting blood, as an alternative. METHODS: HbA(1c) was measured in 1645 children aged 9-11 years without diabetes from the Avon Longitudinal Study of Parents and Children. Thirty-nine children had two HbA(1c) measurements. Data on parental, child and potential confounding factors were collected prospectively from questionnaires, medical records and direct examination. Data from a shortened 30-min oral glucose tolerance test were available for 431 children at age 8 years. Body composition was measured by dual energy X-ray absorptiometry. RESULTS: Mean (sd) HbA(1c) was 4.91(0.29)%. HbA(1c) increased with age and was higher in boys compared with girls, non-white compared with white children, and in children with anaemia. Mean difference between repeated HbA(1c) measurements was 0.01%. HbA(1c) was weakly positively associated with fasting glucose (beta = 0.066%/mmol/l, P = 0.05), but was not associated with 30-min glucose, fasting or 30-min insulin, or homeostasis model assessment insulin resistance. HbA(1c) was weakly inversely associated with weight sd score (beta =-0.02%/unit, P = 0.004), body mass index sd score (beta = -0.02%/unit, P = 0.002), and total body fat (beta = -0.003%/kg, P = 0.06) and lean mass (beta = 0.011%/kg, P = 0.01), but was not associated with birthweight or breastfeeding. CONCLUSIONS: HbA(1c) is not a good marker of fasting or post-load glucose and insulin measures in healthy children, and is not a viable alternative to these measures for investigating the determinants of insulin resistance and Type 2 diabetes in children. PMID- 17116189 TI - Divergent trends in the incidence of end-stage renal disease due to Type 1 and Type 2 diabetes in Europe, Canada and Australia during 1998-2002. AB - AIMS: To describe the variation in geographical distribution of end-stage renal disease (ESRD) due to Type 1 and Type 2 diabetes, and to calculate recent trends in incidence in predominantly white populations. METHODS: Estimation of age- and sex-standardized incidence of ESRD by type of diabetes, and temporal trends, in population-based data for persons aged 30-44, 45-54 or 55-64 years newly treated for ESRD during 1998-2002 in eight countries or regions of Europe, and Non Indigenous Canadians and Australians. RESULTS: The incidence of ESRD due to Type 1 diabetes at age 30-44 years correlated with published rates of childhood-onset insulin dependent diabetes mellitus (P = 0.0025). ESRD due to Type 2 diabetes was uncommon before 45 years of age; in older persons, the highest rates (in Canada and Austria) were five times the lowest rates (in Norway and the Basque region). Rates of ESRD due to Type 1 diabetes fell, per year, by 6.4%[95% confidence interval (CI): 2.1-10.6%) in persons aged 30-44 years, and by 7.7% (95% CI: 2.4 12.7%] in those aged 45-54 years. In contrast, rates of ESRD due to Type 2 diabetes increased annually by 16% (95% CI: 5-28%) in the 30-44-year age group, 11% (95% CI: 6-16%) at 45-54 years, and 9% (95% CI: 5-14%) at 55-64 years. CONCLUSIONS: Modern prevention has reduced progression of nephropathy to ESRD due to Type 1 diabetes, but the continuing rise of ESRD due to Type 2 diabetes represents a failure of current disease control measures that has serious public health implications. PMID- 17116190 TI - Predictive value of admission hyperglycaemia on mortality in patients with acute myocardial infarction. AB - RATIONALE AND AIM: In patients with an acute myocardial infarction, admission hyperglycaemia (AH) is a major risk factor for mortality. However, the predictive value of AH, when the risk score and use of guidelines-recommended treatments are considered, is poorly documented. METHODS: The first fasting plasma glucose levels after admission, risk level, guidelines-recommended treatment use and 1 year mortality were recorded. Patients with first fasting glucose level after admission > 7.7 mmo/l were considered to have AH. RESULTS: Three hundred and twenty patients with ST segment elevation myocardial infarction (STEMI) and 404 with non-ST segment elevation myocardial infarction (NSTEMI) were included. One hundred and seventy-five (24%) patients had pre-existing diabetes (diabetes group), 154 (21%) had AH (AH+ group) and the remainding 395 (55%) had neither diabetes nor AH (AH- group). The Global Registry of Acute Coronary Events (GRACE) risk score was lower in the AH- group, but the use of guidelines-recommended treatment was comparable in all groups. At 1 year, the mortality rate was higher in the AH+ group compared with the AH- group (18.8 vs. 6.1%, P < 0.01) and similar to that in the diabetes group (18.8 vs. 16.6%, P = NS). The relation between glycaemic status and mortality remained strong [AH+ vs. AH-, OR = 3.0 (1.5, 6.0) and diabetes vs. AH-, OR = 3.6 (1.7, 6.6)] after adjustment for the GRACE risk score [OR = 2.4 (1.8, 3.1) per 10% increase] and for treatment score [OR = 0.7 (0.6, 0.8) per 10% increase]. CONCLUSIONS: In patients without a history of diabetes, the presence of AH indicates an increased risk of 1-year mortality, similar to that of patients with diabetes, even when the risk score and use of guidelines-recommended treatment are controlled for. PMID- 17116191 TI - Prevalence of and risk factors for extracranial internal carotid artery stenosis in Korean Type 2 diabetic patients. AB - AIMS: The objectives of this study were to evaluate the prevalence of and risk factors for extracranial internal carotid artery stenosis in Type 2 diabetic patients. METHODS: This study included 406 patients aged 40-79 years with Type 2 diabetes (male 55.4%, female 44.6%). Both carotid arteries of each patient were examined by carotid duplex scanning. The duplex ultrasound criteria based on the North American Symptomatic Carotid Endarterectomy Trial (NASCET) measurement method were used for the identification of carotid stenosis. RESULTS: Extracranial internal carotid artery stenosis >or= 40% by velocity criteria was detected in 5.2% of the patients. The prevalence of carotid stenosis increased with advancing age: 1.0% at 40-49 years of age, 5.0% at 50-59 years, 7.3% at 60 69 years and 9.5% at 70-79 years. The degree of stenosis was > 70% in 42.9% of patients with stenosis, Bilateral stenosis was detected in 14% of patients. Of the patients with >or= 40% carotid stenosis, 33% had a decreased ankle-brachial index, 38% had a previous history of stroke, and only one patient (5%) had a documented history of coronary artery disease. Multivariate analysis, including variables determined to be significantly different by univariate analysis between patients with or without >or= 40% stenosis, indicated that age, systolic blood pressure and high-density lipoprotein (HDL)-cholesterol (inverse correlation) were independent risk factors associated with carotid stenosis. CONCLUSIONS: Carotid duplex scanning is a useful strategy in identifying carotid stenosis in older Type 2 diabetic patients with high systolic blood pressure, or low levels of HDL cholesterol. The early identification and subsequent appropriate management of carotid stenosis, particularly in this group of patients, may facilitate efforts to reduce the incidence of macrovascular complications. PMID- 17116192 TI - The prevalence of depression and anxiety in adults with Type 1 diabetes. AB - AIM: The aim of the present study was to determine the prevalence of anxiety and depression in a large UK group of people with Type 1 diabetes. METHODS: Patients aged 16-60 years were invited to complete self-report questionnaires when they attended outpatient clinic appointments. HbA(1c) was recorded from the clinic database. RESULTS: Analysis was based on 273 complete questionnaires. The mean scores for both anxiety (mean 6.4, sd 4.5) and depression (mean 3.4, sd 3.5) were consistent with normative data. Females reported significantly higher mean anxiety than males, although neither reached the criterion for 'caseness'. Significant differences to the norm were observed for the percentages reporting moderate to severe levels of depression in males (chi(2) = 6.44; d.f. = 2; P = 0.04) and moderate to severe levels of anxiety in females (chi(2) = 7.47; d.f. = 2; P = 0.02). HbA(1c) was positively correlated with HADS scores (anxiety r = 0.2, P = 0.001, depression r = 0.14, P = 0.02). CONCLUSIONS: While there is no significant difference in the mean anxiety or depression in this cohort compared with those reported for a non-diabetic, healthy population, the results suggest that there is an increased prevalence of clinically relevant anxiety in females and of depression in males with Type 1 diabetes when compared with the normative data. PMID- 17116194 TI - Cut points of waist circumference for Japanese people. PMID- 17116195 TI - Rupture of the Achilles tendon associated with ciprofloxacin. PMID- 17116196 TI - Interleukin-1 and the constellation of pulmonary oedema, and cerebral infarctions and oedema in diabetic ketoacidosis. PMID- 17116198 TI - Selective obstruction of lateral striate capsular arteries due to a small cardiogenic embolus as a cause of acute cerebral infarction limited to unilateral putamen. PMID- 17116204 TI - Late onset stroke and myocardial infarction in Williams syndrome. PMID- 17116205 TI - Distal painful peripheral neuropathy associated with erythema induratum of Bazin. PMID- 17116206 TI - Increased levels of coagulation factor VIII in internal carotid artery occlusion. PMID- 17116207 TI - EFNS guidelines on the use of skin biopsy in the diagnosis of peripheral neuropathy. PMID- 17116208 TI - EFNS guideline on neuroimaging in acute stroke. Report of an EFNS task force. AB - Neuroimaging techniques are necessary for the evaluation of stroke, one of the leading causes of death and neurological impairment in developed countries. The multiplicity of techniques available has increased the complexity of decision making for physicians. We performed a comprehensive review of the literature in English for the period 1965-2005 and critically assessed the relevant publications. The members of the panel reviewed and corrected an initial draft, until a consensus was reached on recommendations stratified according to the European Federation of Neurological Societies (EFNS) criteria. Non-contrast computed tomography (CT) scan is the established imaging procedure for the initial evaluation of stroke patients. However, magnetic resonance imaging (MRI) has a higher sensitivity than CT for the demonstration of infarcted or ischemic areas and depicts well acute and chronic intracerebral hemorrhage. Perfusion and diffusion MRI together with MR angiography (MRA) are very helpful for the acute evaluation of patients with ischemic stroke. MRI and MRA are the recommended techniques for screening cerebral aneurysms and for the diagnosis of cerebral venous thrombosis and arterial dissection. For the non-invasive study of extracranial vessels, MRA is less portable and more expensive than ultrasonography but it has higher sensitivity and specificity for carotid stenosis. Transcranial Doppler is very useful for monitoring arterial reperfusion after thrombolysis, for the diagnosis of intracranial stenosis and of right-to left shunts, and for monitoring vasospasm after subarachnoid hemorrhage. Currently, single photon emission computed tomography and positron emission tomography have a more limited role in the evaluation of the acute stroke patient. PMID- 17116209 TI - PECAM-1, a key player in neuroinflammation. AB - Platelet endothelial cell adhesion molecule-1 (PECAM-1, CD31) is a 130-kDa protein, which plays a significant role in the adhesion cascade. It is therefore involved in leucocyte endothelium interaction and in leucocyte transendothelial migration during inflammation. As neuroinflammation and subsequent blood brain barrier disruption are integral processes in many neurological disorders, PECAM-1 and its soluble form (sPECAM-1) have been investigated in a number of conditions, rising hopes as a potential marker of disease activity, a possible target in treatment and a prognostic factor. It has been shown that serum and CSF levels of PECAM-1 and sPECAM-1 are increased in patients in active stages of multiple sclerosis. Similarly, they rise in individuals after ischaemic stroke. PECAM-1 has also been shown to be involved in the pathogenesis of Abeta-related cerebral vascular disorders, such as Alzheimer disease. It participates in the pathomechanism of paraneoplastic neurological disorders and in neuroinflammation in NeuroAIDS. A number of experiments on animal models were carried out in order to investigate PECAM-1 role in the above-mentioned conditions and more, including brain trauma and nerve root injury. In this review most recent investigations on PECAM-1 biology and its role in neuroinflammation have been described and discussed from a multidisciplinary point of view. PMID- 17116210 TI - Overactive bladder in Parkinson's disease: alteration of brainstem raphe detected by transcranial sonography. AB - Urinary dysfunction is very common in idiopathic Parkinson's disease (PD) and manifests primarily with symptoms of overactive bladder (OAB). Affection of central serotonergic systems has been suggested to play a role in OAB. The objective of this study was to evaluate whether in PD patients with OAB symptoms a specific alteration of the brainstem raphe (BR), which contains serotonergic neurons, can be detected with transcranial sonography (TCS). Of 116 PD patients enrolled, 19 had PD-related OAB symptoms (OAB+) unlike remaining 97 patients (OAB ). Patients were examined by a sonographer blinded to the clinical data. Reduced echogenicity of BR was found in 12 (63%) OAB+ patients but only in 18 (19%) of 93 assessable OAB- patients (Mann-Whitney U-test, P < 0.001). In OAB+ patients, lower raphe echogenicity score was associated with longer duration of OAB symptoms (anova, P = 0.033). Other TCS findings such as echogenicity of substantia nigra, thalami, lenticular and caudate nuclei, and widths of third and lateral ventricles did not differ between OAB+ and OAB- patients. TCS findings suggest a pathogenetic role of BR in OAB related to PD. Alterations may reflect disturbance of its central serotonergic system. PMID- 17116211 TI - LRRK2 mutations in a clinic-based cohort of Parkinson's disease. AB - In the last decade, major breakthroughs in the understanding of genetic contributions to Parkinson's disease (PD) have been achieved. Recently, mutations in LRRK2, encoding dardarin, have been found to be responsible for an autosomal dominant parkinsonism (OMIM 607060). We screened 311 subjects (cases: n = 202, controls: n = 109) for the three previously reported LRRK2 mutations. Our investigation revealed a sporadic case of PD with a heterozygous mutation G2019S (c.6055G>A). Here, we present the clinical phenotype of this patient and discuss the implications of genetic testing for the G2019S mutation in patients with sporadic PD. PMID- 17116212 TI - Clinical features of childhood onset essential tremor. AB - Childhood onset essential tremor (ET) is uncommon. It is not clear as to whether ethnicity-specific differences may influence the phenotypic features. To determine the frequency and clinical characteristics of childhood ET in a tertiary referral center. In a prospective evaluation of 120 consecutive ET patients in a movement disorders clinic, we found a 15.5% (19) frequency of childhood onset ET patients. The mean age of onset and mean age was 10.8 +/- 4.1 (6-16) years and 25.7 +/- 15.0 (16-73) years consisting of 73.6% (14/19) men and 26.4% (5/19) women. A positive family history of ET was present in 11 of 19 (52.6%). Presence of a head tremor was observed in 2/19 (10.5%). We highlighted a relatively high frequency (15,5%) of childhood ET in our Asian cohort. In addition, we drew attention to the male preponderance and the low frequency of head tremor in childhood ET corroborating study findings in white ET patients. These observations appear to transcend ethnic and cultural differences and lend further support that gender difference may play a role in the pathogenesis and expression of ET. PMID- 17116213 TI - Long-term treatment with pramipexole in restless legs syndrome. AB - The aim of the present study was to look at the long-term efficacy and side effects profiles of pramipexole in a large cohort of drug naive patients with regard to dopaminergic medications. In all, 195 consecutive restless legs syndrome (RLS) patients who were prescribed pramipexole more than 1 year previously, agreed to undergo a telephone interview to assess both the efficacy and side effects of pramipexole. Forty-three patients had discontinued pramipexole: 20 because of side effects, six because of a lack of efficacy, six for both and 11 for other reasons. Patients who continued pramipexole for more than 1 year (n = 152) reported a mean decrease in RLS symptoms severity of 80.9% (SD = 19.6%). At the onset of treatment, the most common side effects were nausea (30%), tiredness (9%), dizziness (8%), headache (4%), insomnia (3%), dry mouth (2%), difficulty to concentrate (1.3%) and sleepiness (0.7%), At 30 months, most patients (n = 124/152; 81.6%) reported an absence of side effects of pramipexole. None of the adverse effects occurred in more than 5% of patients at follow-up. The present study confirms, in a large cohort of unselected patients, that pramipexole is effective and safe in the long-term treatment of RLS. PMID- 17116214 TI - Symmetry of post-movement beta-ERS and motor recovery from stroke: a low resolution EEG pilot study. AB - The inter-hemispheric symmetry of electroencephalographic (EEG) post-movement beta-event-related synchronization (PMBS) after movements on a drawing board was studied in eight acute stroke subjects with mild hemiparesis and eight normal subjects. A follow-up testing was conducted 3 months after the initial recordings with a twofold purpose: (1) to validate the reproducibility of the experimental protocol in normal subjects; and (2) to study changes of inter-hemispheric PMBS symmetry as a response to recovery of motor function. PMBS values were calculated and their topographic distributions illustrated at various time instances following movement offset. Significant PMBS patterns were present in all normal subjects, with only minor differences within consecutive recordings. The side of hemiparesis in acute stroke subjects could be distinguished (P = 0.04) on the basis of the signed symmetry index, a quantitative measure of lateralization. The follow-up testing on three recovered stroke subjects revealed a trend of changes in the lateralization towards the contralateral side of movement, an indication that the cortical organization of movement following recovery turned out as reported for normal subjects. Further clinical investigations need to be carried out to evaluate the relationship between recovery and PMBS symmetry on a large number of subjects, using the method presented here. PMID- 17116215 TI - Gene expression profiling in the human middle cerebral artery after cerebral ischemia. AB - We have investigated the gene expression in human middle cerebral artery (MCA) after ischemia. Ischemic stroke affects the perfusion in the affected area and experimental cerebral ischemia results in upregulation of vasopressor receptors in the MCA leading to the ischemic area. We obtained human MCA samples distributing to the ischemic area, 7-10 days post-stroke. The gene expression was examined with real-time polymerase chain reaction (PCR) and microarray, proteins were studied with immunohistochemistry. We investigated genes previously shown to be upregulated in animal models of cerebral ischemia (e.g. ET(A), ET(B), AT1, AT2, and 5-HT(2A/1B/1D)). Their mRNA expression was increased compared with controls, consistent with findings in experimental stroke. Immunohistochemistry showed upregulation of the receptors localized on the smooth muscle cells. The gene expression was profiled with microarray and seven genes chosen for further investigation with real-time PCR; ELK3, LY64, Metallothionin IG, POU3F4, Actin alpha2, RhoA and smoothelin. Six of these were regulated the same way when confirming array expression with real-time PCR. Gene expression studies in the human MCA leading to the ischemic region is similar to that seen after MCA occlusion in rats. We found new genes that support the dynamic changes that occur in the MCA distributing to the ischemic region. PMID- 17116216 TI - Fatigue and sleep disturbance in multiple sclerosis. AB - Considering the association of sleep disturbance and fatigue in multiple sclerosis (MS), we investigated the presence of sleep disturbances that may be related to fatigue by using objective and subjective measures. We included 27 MS patients with fatigue, 10 MS patients without fatigue and 13 controls. The Pittsburgh sleep quality index score showed significant differences between patient groups and controls. Beck depression inventory scores were significantly higher in fatigued than non-fatigued patients. Comparison of patient groups and controls revealed significant differences for time in bed, sleep efficiency index, sleep continuity index, wake time after sleep onset, total arousal index and periodic limb movement arousal index. Our study confirms that MS causes sleep fragmentation in terms of both macro and microstructure. Fatigue in MS could be partially explained by disruption of sleep microstructure, poor subjective sleep quality and depression. PMID- 17116217 TI - Comparison of the growth hormone, IGF-1 and insulin in cerebrospinal fluid and serum between patients with motor neuron disease and healthy controls. AB - Neurotrophic effects of the growth hormone (GH), insulin-like growth factor-1 (IGF-1) and insulin on the central nervous system have become more apparent in the past decade. In this study, we measured serum and cerebrospinal fluid (CSF) concentrations of GH, IGF-1 and insulin in 35 patients with motor neuron disease (MND) [24 patients with definite amyotrophic lateral sclerosis (ALS) and 11 patients with progressive bulbar palsy] and in 40 healthy controls. Levels of serum concentrations of GH and IGF-1 did not significantly differ between the MND patient group and the healthy controls, while the level of insulin was significantly decreased (P = 0.0033) in the MND patient group. However, levels of all three examined parameters in CSF were significantly lower in the MND group than in the healthy controls with the statistical significance for IGF-1 and insulin of P < 0.001. This finding has not been reported previously, and further investigations into its association with ALS should establish whether it can be used as an early marker of the disease, or whether it merely represents a consequence of ALS development. PMID- 17116218 TI - Increased urinary excretion of nitric oxide metabolites in longitudinally monitored migraine patients. AB - This study evaluated a relationship between nitric oxide (NO) and migraine attacks in order to gain insight into migraine pathomechanism. The study groups consisted of 12 migraineurs and eight controls. All subjects collected morning urine samples for 40 consecutive days. Urinary NO metabolites, nitrite/nitrate (NO(x)) levels were measured with the vanadium-based assay, whilst creatinine (Cr) and neopterin were determined with high-performance liquid chromatography. The mean urinary NO(x)/Cr ratio and number of NO(x) peaks was significantly greater in the migraine group compared with controls (P = 0.01 and P = 0.007, respectively). In the second approach, high NO(x) values were re-assessed in relation to raised neopterin, a marker of systemic infection or inflammation, and were excluded. The excretion of NO(x) persisted being pulsatile, and migraineurs had more peaks compared with controls (P = 0.01). In seven patients, NO(x) peaks coincided with headache days. This was more frequent than expected by random association in four patients (Monte-Carlo simulation; odds ratios: 2.16-7.77; no overlap of 95% CI). In four patients, NO(x) peaks preceded or followed headache days. Although there is a difference in the pattern of urinary NO(x) excretion between control and migraine populations, the variable temporal association of NO(x) peaks and headaches suggests a complex role of NO in this condition. PMID- 17116219 TI - Influence of levetiracetame on ictal and postictal EEG in patients with partial seizures. AB - To investigate the influence of levetiracetame (LEV) treatment on the interhemispheric seizure pattern propagation and postictal recovery of electroencephalography (EEG) background activity. Twenty-three adult patients (age > 16 years) with pharmacoresistant focal epilepsies presenting at the Epilepsy Center Erlangen for pre-surgical evaluation were enrolled in the study. Those eligible patients receiving only one antiepileptic drugs were recruited to the 48-h baseline phase and, after at least two seizures, were randomized to the 7-day treatment phase with either LEV (n = 11) or placebo (n = 12). All participants were submitted to continuous day-and-night video-EEG monitoring. The daily dose of LEV was 1000 mg (500 mg bid.) on the first treatment day and was increased to 2000 mg (1000 mg bid.) from the second day onward. The EEG changes relating to the time delay of the interhemispheric seizure pattern propagation and to the postictal recovery of the background activity were analysed by computerized video-EEG recording and compared using the non-parameter Mann Whitney U-exact test (alpha = 0.05). A prolonged latency of the contralateral seizure pattern propagation was observed in the LEV group, whereas a more rapid propagation was observed in the placebo group (P = 0.009). Postictal generalized slowing of the background activity was recorded in 21 patients during the baseline phase. More rapid postictal recovery of the EEG background activity was observed in the LEV, but not in the placebo group (P = 0.03). This study demonstrated that LEV not only prevented the seizure pattern propagation but also helped the speedy recovery of the postictal background activity in the EEG. PMID- 17116220 TI - Expression of chemokine receptors on peripheral blood mononuclear cells of patients with immune-mediated neuropathies treated with intravenous immunoglobulins. AB - Intravenous immunoglobulin (IVIg) is an efficacious treatment for immune-mediated neuropathies like Guillain-Barre syndrome (GBS), chronic inflammatory demyelinating neuropathy (CIDP), and multifocal motor neuropathy (MMN). In the pathogenesis of immune-mediated neuropathies chemokines and their receptors play a crucial role. Using flow cytometry we examined whether IVIg modulates chemokine expression repertoires of T cells and monocytes. The expression of inflammatory chemokine receptors CCR1, CCR2, CCR4, CCR5, CCR6 and CXCR3 was investigated on circulating T-cell subsets, and CCR1, CCR2 and CCR5 on circulating monocytes before and after IVIg treatment in patients with immune-mediated neuropathies (MMN, n = 7; GBS, n = 1; CIDP, n = 2). Furthermore, the homing potential of T cells was analyzed by the expression of CCR7, a chemokine receptor known to be utilized by mature T cells to recirculate into secondary lymphoid organs. In contrast to studies in chronic heart failure, no differences in expression patterns before and after IVIg treatment of any of the investigated chemokine receptors were found. Furthermore, the proportion of CD45RO-positive CD4+ or CD8+ T-cell subsets was not changed by IVIg treatment. Thus, we concluded that modulation of the expression of chemokine receptors on circulating leukocytes by IVIg is not a mode of action in immune-mediated neuropathies. PMID- 17116221 TI - Brain MRI white matter lesions in migraine patients: is there a relationship with antiphospholipid antibodies and coagulation parameters? AB - Brain magnetic resonance imaging (MRI) studies in migraine patients have demonstrated lesions consisting of focal regions of increased signal intensity within the white matter. Antiphospholipid antibodies are known to have a role in many diseases including migraine. The aim of the present study was to ascertain the relationship between MRI-visualized cerebral focal hyperintense lesions and serum antiphospholipid antibody levels, as well as blood coagulation parameters in migraine patients. One hundred and two (77 females, 25 males, mean age 33.8 +/ 11.1) consecutive migraine patients and a control group of 94 (70 females, 24 males, mean age 33.2 +/- 10.8) healthy subjects were enrolled. All individuals underwent brain MRI. Complete blood examinations, autoantibodies, antiphospholipids antibodies including anticardiolipin and lupus anticoagulant (aCL, LAC), antithrombin III, Protein C and S serum levels were ascertained in the subjects who presented white matter lesions on MRI. Twenty-seven (26.4%) migraine patients and six (6.3%) healthy subjects in the control group showed focal regions of increased intensity signal within cerebral white matter (odds ratio 5.3, 95% CI: 1.98-16.36). In migraine patients with white matter lesions, antiphospholipid antibodies were not detected and serum levels of antithrombin III, and proteins C and S were normal. White matter lesions in migraine patients are fairly common. This finding is not associated with antiphospholipid antibodies or abnormal coagulation parameters. The significance of such lesions at present remains unclear. PMID- 17116222 TI - Is uncertain diagnosis a more frequent reason for referring migraine patients to neurologist than other headache syndromes? AB - The objective was to assess whether the physician's uncertainty of diagnosis was a more frequent motivation for referring migraine patients than other headache patients to a neurologist. In this population based survey we included 846 consecutive patients referred to and examined at a specialist centre for headache during a period of 2 years. As primary outcome we compared cross-sectionally frequency of referring patients with migraine and other headaches to neurologist. According to the patients, uncertainty of diagnosis was the main reason for referral to a neurological specialist consultation, i.e. in 51% of cases. Thirty nine per cent of patients classified as having migraine were referred because of an uncertain diagnosis, compared with 62% in patient with other headaches (95 CI 0.13-0.33), (P < 0.0001). Headache subtypes other than migraine and younger age were associated to more uncertainty of diagnosis. We conclude that diagnostic uncertainty was an important factor amongst half the headache patients seen by neurologists and uncertain diagnoses as reason for referral was more important amongst patients with non-migrainous headache. PMID- 17116223 TI - The number of pregnancies is a risk factor for Alzheimer's disease. AB - Epidemiological data show a higher prevalence of late-onset Alzheimer's disease (AD) in women. The estrogenic deficiency in the post-menopausal period is suspected to be the cause of the gender-related risk of the disease, but studies on the estrogenic therapy and occurrence of AD were not consistent and sometimes contradicting. The aim of this study is to investigate whether a higher exposure to endogenous estrogens is associated with lower risk of dementia or not. Two hundred and four AD patients and 201 control women were considered. By interviews, we evaluated different variables, indirectly correlated to estrogenic natural exposure, as well as educational level and head trauma. These data were correlated in the AD group with the disease progression, as well as with the age at onset. Unexpectedly, we found a significant higher number of pregnancies in the AD than in the control group. Within the AD cases, the number of lifetime pregnancies is related to an earlier onset of the disease. As previously reported, we confirmed that the educational level is a protective factor and that major head trauma represents a risk factor in developing AD. The higher number of pregnancies and a less frequency of nulliparous women, indirectly relate the AD group to a higher estro-progestinic exposure. These findings suggest that it is the increase of progesterone or estrogens level--and not the estrogens decrease, as previously indicated by other authors--that could play a role in the Alzheimer's pathology. PMID- 17116224 TI - Abnormal musical pacemaker in a patient with musical hallucinations. AB - Music creation requires a highly orchestrated temporal pattern. The study of a patient with repetitive musical hallucinations enabled us to examine temporal pacemakers in music production. Here, we show that the pattern of faster silent production of a chosen tune compared with its production aloud was reversed when the patient produced the hallucinatory tune. This observation might suggest the utilization of a pacemaker(s), which functions differentially during the disease. PMID- 17116225 TI - Dissociated small hand muscle atrophy in aging: the 'senile hand' is a split hand. AB - The term 'split hand' refers to a pattern of dissociated atrophy of hand muscles and was first described in ALS. We hypothesize that this phenomenon also occurs in 'normal' aging. We investigated healthy subjects of different ages and found a progressive dissociation in atrophy of the hand muscles, as measured with compound muscle action potential amplitudes, with increasing age. Different possible causes of this progressive dissociation are discussed. It might be related to preferential use of thenar muscles in humans, which render these muscles and their motor neurons more susceptible to oxidative stress. In addition, a difference in intrinsic susceptibility to oxidative stress might be involved. The relation between normal age-related muscle loss (sarcopenia) and the pathologic loss in motor neuron disease is discussed. PMID- 17116226 TI - High-fat diet impairs hippocampal neurogenesis in male rats. AB - High fat diets and obesity pose serious health problems, such as type II diabetes and cardiovascular disease. Impaired cognitive function is also associated with high fat intake. In this study, we show that just 4 weeks of feeding a diet rich in fat ad libitum decreased hippocampal neurogenesis in male, but not female, rats. There was no obesity, but male rats fed a diet rich in fat exhibited elevated serum corticosterone levels compared with those fed standard rat chow. These data indicate that high dietary fat intake can disrupt hippocampal neurogenesis, probably through an increase in serum corticosterone levels, and that males are more susceptible than females. PMID- 17116227 TI - Preliminary report on effects of oxcarbazepine-treatment on serum lipid levels in children. AB - The aim of the present study was to assess serum lipid levels before and after treatment with oxcarbazepine (OXC) in children with epilepsy. We measured total cholesterol (TC), triglycerides (TGs) and high-density lipoprotein cholesterol (HDL-C) in 28 patients whereas only TC levels in 11 patients, during baseline period and at 3 months after the beginning of therapy with OXC. During baseline period, median values were: 4.38 mmol/l (IQR = 4.12-5.03) for TC levels, 1.72 mmol/l (IQR = 1.42-2.01) for HDL-C levels and 1.54 mmol/l (IQR = 1.29-1.96) for TGs levels. At 3 months, median values were: 4.38 mmol/l (4.10-4.95) for TC levels (P < 0.05), 1.57 mmol/l (1.34-1.93) for HDL-C levels (P < 0.005) and 1.8 mmol/l (1.23-2.34) for TGs levels (P < 0.05). Median serum lipid levels remained in the normal range, despite an increasing-trend at 3 months of treatment with OXC. Further studies are necessary to confirm these results. PMID- 17116228 TI - Brain-derived neurotrophic factor regulates the expression of D1 dopamine receptors. AB - We have previously demonstrated that the CAD catecholaminergic neuronal cell line is an appropriate model system to study the regulation of D(1) dopamine receptor expression. In this report, we show that brain-derived neurotrophic factor (BDNF) up-regulates the expression of D(1) dopamine receptor in CAD cells. In addition, by comparing D(1) receptor mRNA expression in wild-type, heterozygous and homozygous trkB knockout mice, we show that TrkB receptor signaling up-regulates D(1) receptor expression in vivo. In CAD cells expressing the TrkB receptor, BDNF increased D(1) receptor mRNA in a time- and dose-dependent manner with a fourfold increase in D(1) receptor mRNA observed as early as 3 h with 10 ng/mL of BDNF. Using different classes and concentrations of kinase inhibitors, we determined that BDNF-induced increase of D(1) receptor mRNA is mediated by the phosphatidylinositol 3-kinase signaling pathway. The increase required both new transcription and protein synthesis, as it was blocked by actinomycin D and cyclohexamide, respectively. Promoter deletion analysis identified a D(1) promoter region necessary for mediating the effect of BDNF. These results provide novel evidence that D(1) dopamine receptor expression is regulated by BDNF and its signaling pathway. PMID- 17116229 TI - Nitric oxide regulates cell survival in purified cultures of avian retinal neurons: involvement of multiple transduction pathways. AB - Nitric oxide (NO) is an important signaling molecule in the CNS, regulating neuronal survival, proliferation and differentiation. Here, we explored the mechanism by which NO, produced from the NO donor S-nitroso-acetyl-d-l penicillamine (SNAP), exerts its neuroprotective effect in purified cultures of chick retinal neurons. Cultures prepared from 8-day-old chick embryo retinas and incubated for 24 h (1 day in culture, C1) were treated or not with SNAP, incubated for a further 72 h (up to 4 days in culture, C4), fixed, and the number of cells estimated, or processed for cell death estimation, by measuring the reduction of the metabolic dye 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT). Experimental cultures were run in parallel but were re-fed with fresh medium in the absence or presence of SNAP at culture day 3 (C3), incubated for a further 24 h up to C4, then fixed or processed for the MTT assay. Previous studies showed that the re-feeding procedure promotes extensive cell death. SNAP prevented this death in a concentration- and time-dependent manner through the activation of soluble guanylate cyclase; this protection was significantly reversed by the enzyme inhibitors 1H-[1,2,4]oxadiazolo-[4,3 a]quinoxalin-1-one (ODQ) or LY83583, and mimicked by 8-bromo cyclic guanosine 5' phosphate (8Br-cGMP) (GMP) or 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole (YC 1), guanylate cyclase activators. The effect was blocked by the NO scavenger 2 phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO). The effect of NO was also suppressed by LY294002, Wortmannin, PD98059, KN93 or H89, indicating the involvement, respectively, of phosphatidylinositol-3 kinase, extracellular regulated kinases, calmodulin-dependent kinases and protein kinase A signaling pathways. NO also induced a significant increase of neurite outgrowth, indicative of neuronal differentiation, and blocked cell death induced by hydrogen peroxide. Cyclosporin A, an inhibitor of the mitochondrial permeability transition pore considered an important mediator of apoptosis and necrosis, as well as boc aspartyl (OMe) fluoromethylketone (BAF), a caspase inhibitor, also blocked cell death induced by re-feeding the cultures. These findings demonstrate that NO inhibits apoptosis of retinal neurons in a cGMP/protein kinase G (PKG)-dependent way, and strengthens the notion that NO plays an important role during CNS development. PMID- 17116230 TI - Expression and evolution of the mammalian brain gene Ttyh1. AB - Homologues of the Drosophila melanogaster tweety (tty) gene are present in mammals and Caenorhabditis elegans. The encoded proteins have five predicted membrane-spanning regions and recent findings suggest that some family members may be chloride channels. Phylogenetic analysis of the tty family including novel members from slime mould Entamoeba and plants has revealed the occurrence of independent gene duplication events in different lineages. expressed sequence tag data indicate that expression of the mammalian Ttyh1 gene is restricted mainly to neural tissue and is up-regulated in astrocytoma, glioma and several other cancers. In this study, mammalian expression vectors were used to investigate the subcellular localization and the effect of over-expression of Ttyh1 in human epithelial kidney cells. The results confirm that Ttyh1 is a membrane protein and show that it is deposited on the substratum along the migration paths of motile cells above the alpha5beta1-integrin complex. The ectopic expression of Ttyh1 also induced long filopodia, which were branched and dynamic in both stationary and migratory cells. The filopodia contained F-actin and occurred at the ends of microtubules which were polarized towards the membrane. Upon contact with nearby cells some filopodia stabilized and filled with F-actin, whereas Ttyh1 was highly concentrated at the cell-cell interface. Ttyh1 N- and C-terminal antipeptide antibodies detected Ttyh1 along the axons of neurones in primary rat hippocampal cell cultures, and in situ in whole rat brain slices around the hippocampus and occasionally between cells. These data suggest a role for Ttyh1 in process formation, cell adhesion and possibly as a transmembrane receptor. PMID- 17116231 TI - Induction of haem oxygenase-1 causes cortical non-haem iron increase in experimental pneumococcal meningitis: evidence that concomitant ferritin up regulation prevents iron-induced oxidative damage. AB - Desferrioxamine inhibits cortical necrosis in neonatal rats with experimental pneumococcal meningitis, suggesting that iron-induced oxidative damage might be responsible for neuronal damage. We therefore examined the spatial and temporal profile of changes in cortical iron and iron homeostatic proteins during pneumococcal meningitis. Infection was associated with a steady and global increase of non-haem iron in the cortex, particularly in neuronal cell bodies of layer II and V, and in capillary endothelial cells. The non-haem iron increase was associated with induction of haem oxygenase (HO)-1 in neurones, microglia and capillary endothelial cells, whereas HO-2 levels remained unchanged, suggesting that the non-haem iron increase might be the result of HO-1-mediated haem degradation. Indeed, treatment with the haem oxygenase inhibitor tin protoporphyrin (which completely blocked the accumulation of bilirubin detected in HO-1-positive cells) completely prevented the infection-associated non-haem iron increase. The same cells also displayed markedly increased ferritin staining, the increase of which occurred independently of HO activity. At the same time, no increase in DNA/RNA oxidation was observed in infected animals (as assessed by in situ detection of 8-hydroxy[deoxy]guanosine), strongly suggesting that ferritin up-regulation protected the brain from iron-induced oxidative damage. Thus, although pneumococcal meningitis leads to an increase of cortical non-haem iron, protective mechanisms up-regulated in parallel prevent iron induced oxidative damage. Cortical damage does not appear to be a direct consequence of increased iron, therefore. PMID- 17116232 TI - Striatal susceptibility to a dopaminergic neurotoxin is independent of sex hormone effects on cell survival and DAT expression but is exacerbated by central aromatase inhibition. AB - The aim of this study was to investigate further the hormone-dependent processes underlying sex differences in neurotoxic responses within the rat nigrostriatal dopaminergic (NSDA) pathway after partial lesioning with 6-OHDA, a state thought to mimic the early stages of Parkinson's disease where, in humans and animal models alike, males appear to be more susceptible. Contrary to our hypotheses, hormone manipulations (gonadectomy +/- oestrogen or androgen treatment) failed to alter survival of tyrosine hydroxylase immunoreactive cells in the substantia nigra pars compacta (SNc) after lesioning; this indicates that, unlike inherent sex differences in toxin-induced striatal dopamine depletion, sex differences in cell loss were not hormonally generated, and that hormone-dependent changes in dopamine depletion can occur independently of cell survival. In addition, hormonally induced changes in striatal expression of the dopamine transporter (DAT), an important factor for 6-OHDA toxicity, did not correlate with hormonal influences on striatal dopamine loss and, in males, central inhibition of aromatase prior to 6-OHDA infusion exacerbated striatal dopamine loss with no effect on SNc tyrosine hydroxylase-immunoreactive survival, suggesting locally generated oestrogen is neuroprotective. These results support the novel view that sex steroid hormones produced peripherally and centrally play a significant, sex specific role within the sexually dimorphic NSDA pathway to modulate plastic, compensatory responses aimed at restoring striatal dopamine functionality, without affecting cell loss. PMID- 17116233 TI - Emerging concepts: novel integration of in vivo approaches to localize the function of nicotinic receptors. AB - Nicotinic acetylcholine receptors (nAChRs) are important targets of the neuromodulator acetylcholine (ACh) and the drug nicotine. The role of their different subunits has been analysed for a decade by the creation of knock-out (KO) mice using homologous recombination. This technique shows that a given subunit is necessary for a given function. However, for ubiquitously expressed genes, it cannot demonstrate the localization for a given subunit in which its expression is sufficient, especially for behavioural phenotypes. Sufficient in this context means that the brain region requiring the expression of the gene product has been localized. Novel strategies have therefore been developed to re express, region specifically, nAChR subunits on a KO background using lentiviral vectors. Localized regeneration of fully functional high-affinity nAChRs in defined brain regions has proven that these receptors are sufficient to restore a variety of functions: nicotine-induced dopamine release, nicotine self administration in mice, dopamine neuron firing patterns, and exploratory and locomotor behaviours in a sequential locomotor task testing executive function were thus defined as depending exclusively on the 'knock-back' of beta2*-nAChRs into the ventral tegmental area. These analyses highlight the important role of endogenous cholinergic regulation of a variety of functions. The novel integrated use of restricted re-expressed nAChR subunits with in vivo electrophysiology and automated quantitative behavioural analysis enables the further analysis of defined neuronal circuits in nicotine addiction and higher cognitive function. PMID- 17116234 TI - Identification and characterization of novel activity-dependent transcription factors in rat cortical neurons. AB - Using gene chip analyses, we have identified novel neuronal activity-dependent genes. Application of 25 mM KCl to mature (14-day culture) rat cortical neurons resulted in more than 1.5-fold induction of 19 genes and reduction of 42 genes among 1200 neural genes. Changes in the overall gene expression profiles appeared to be related to the reduction of excitability and induction of cellular survival signals. Among the genes identified, three transcriptional modulators [encoding Cbp/p300-interacting transactivator with ED-rich tail 2 (CITED2), CCAAT/enhancer binding protein beta (C/EBPbeta) and neuronal orphan receptor-1, (NOR1)] were newly identified as activity-dependent transcription factors, and two of these (CITED2 and NOR1) were found to be influenced by electroconvulsive shock (ECS). NOR1 was induced in specific brain regions by behavioral activation, such as exposure to a novel environment. Because the brain regions that exhibited the induction of these newly identified neuronal activity-dependent transcriptional modulators were distinct from those showing the induction of previously identified activity-dependent genes such as c-fos, these genes might be useful markers for mapping neuronal activity in vivo. PMID- 17116235 TI - Monoclonal antibodies that target pathological assemblies of Abeta. AB - Amyloid beta (Abeta) immunotherapy for Alzheimer's disease has shown initial success in mouse models of Alzheimer's disease and in human patients. However, because of meningoencephalitis in clinical trials of active vaccination, approaches using therapeutic antibodies may be preferred. As a novel antigen to generate monoclonal antibodies, the current study has used Abeta oligomers (amyloid beta-derived diffusible ligands, ADDLs), pathological assemblies known to accumulate in Alzheimer's disease brain. Clones were selected for the ability to discriminate Alzheimer's disease from control brains in extracts and tissue sections. These antibodies recognized Abeta oligomers and fibrils but not the physiologically prevalent Abeta monomer. Discrimination derived from an epitope found in assemblies of Abeta1-28 and ADDLs but not in other sequences, including Abeta1-40. Immunoneutralization experiments showed that toxicity and attachment of ADDLs to synapses in culture could be prevented. ADDL-induced reactive oxygen species (ROS) generation was also inhibited, establishing this response to be oligomer-dependent. Inhibition occurred whether ADDLs were prepared in vitro or obtained from Alzheimer's disease brain. As conformationally sensitive monoclonal antibodies that selectively immunoneutralize binding and function of pathological Abeta assemblies, these antibodies provide tools by which pathological Abeta assemblies from Alzheimer's disease brain might be isolated and evaluated, as well as offering a valuable prototype for new antibodies useful for Alzheimer's disease therapeutics. PMID- 17116236 TI - More on: 'new light on an old story: von Willebrand factor binding to collagen'. PMID- 17116237 TI - The risk of venous thrombosis associated with a high endogenous thrombin potential in the absence and presence of activated protein C. PMID- 17116238 TI - Inverse relationship between the severity of gingivitis and platelet reactivity in stable angina pectoris. PMID- 17116239 TI - The novel Hha/YmoA family of nucleoid-associated proteins: use of structural mimicry to modulate the activity of the H-NS family of proteins. AB - The Hha/YmoA family of proteins is a group of conserved, low-molecular-weight proteins involved in the regulation of gene expression. Studies performed in Escherichia coli, Salmonella sp. and Yersinia sp. highlight the contribution of these proteins in regulating bacterial virulence, horizontal gene transfer and cell physiology. Genes encoding such proteins are located on chromosomes and plasmids in different genera of Gram-negative bacteria. Their mode of action is currently being analysed by studying direct binding of Hha to DNA and as a component of protein complexes with regulatory functions. Recent data on the interaction of Hha with the H-NS family of proteins and structural information suggest a physiological role for such protein complexes in many aspects of gene regulation. PMID- 17116240 TI - A membrane-bound FtsH protease is involved in osmoregulation in Synechocystis sp. PCC 6803: the compatible solute synthesizing enzyme GgpS is one of the targets for proteolysis. AB - Protein quality control and proteolysis are involved in cell maintenance and environmental acclimatization in bacteria and eukaryotes. The AAA protease FtsH2 of the cyanobacterium Synechocystis sp. PCC 6803 was identified during a screening for mutants impaired in osmoregulation. The ftsH2(-) mutant was salt sensitive because of a decreased level of the osmoprotectant glucosylglycerol (GG). In spite of wild type-like transcription of the ggpS gene in ftsH2(-) cells the GgpS protein content increased but only low levels of GgpS activity were observed. Consequently, salt tolerance of the ftsH2(-) mutant decreased while addition of external osmolyte complemented the salt sensitivity. The proteolytic degradation of the GgpS protein by FtsH2 was demonstrated by an in vitro assay using inverted membrane vesicles. The GgpS is part of a GG synthesizing complex, because yeast two-hybrid screens identified a close interaction with the GG phosphate phosphatase. Besides GgpS as the first soluble substrate of a cyanobacterial FtsH protease, several other putative targets were identified by a proteomic approach. We present a novel molecular explanation for the salt sensitive phenotype of bacterial ftsH(-) mutants as the result of accumulation of inactive enzymes for compatible solute synthesis, in this case GgpS the key enzyme of GG synthesis. PMID- 17116241 TI - Uncoupling of choline-O-sulphate utilization from osmoprotection in Pseudomonas putida. AB - The genomic context of the recognized bet genes for choline-O-sulphate (COS) utilization in Pseudomonas putida KT2440 is such that betC (choline sulphatase) lies adjacent to an ATP-binding cassette transporter and a LysR type regulator, but well away from betBA, encoding enzymes for transformation of choline into glycine betaine. The consequences of such genetic layout of the functions for COS metabolism have been examined with a suite of genetic and biochemical approaches. An early clue of the utilities of the betencoded products was exposed by the phenotypes of a betC deletion. This mutant still accumulated intact COS but failed to use this compound as carbon or nitrogen source. Furthermore, betC expression was downregulated at high salt concentrations, showing that the principal role of this gene lied in COS metabolism, not in osmoprotection. In contrast, the betBA genes were required for choline transformation into the highly effective compatible solute glycine betaine (and the concomitant endurance to high salt) and also for its utilization as carbon or nitrogen source. Thus, unlike in the cases of Bacillus subtilis and Sinorhizobium meliloti, betC is unrelated to osmoprotection in Pseudomonas putida while the betBA genes are required for both betaine synthesis and tolerance to high osmotic pressure. PMID- 17116242 TI - Overexpression of putative topoisomerase 6 genes from rice confers stress tolerance in transgenic Arabidopsis plants. AB - DNA topoisomerase 6 (TOP6) belongs to a novel family of type II DNA topoisomerases present, other than in archaebacteria, only in plants. Here we report the isolation of full-length cDNAs encoding putative TOP6 subunits A and B from rice (Oryza sativa ssp. indica), preserving all the structural domains conserved among archaebacterial TOP6 homologs and eukaryotic meiotic recombination factor SPO11. OsTOP6A1 was predominantly expressed in prepollinated flowers. The transcript abundance of OsTOP6A2, OsTOP6A3 and OsTOP6B was also higher in prepollinated flowers and callus. The expression of OsTOP6A2, OsTOP6A3 and OsTOP6B was differentially regulated by the plant hormones, auxin, cytokinin, and abscisic acid. Yeast two-hybrid analysis revealed that the full-length OsTOP6B protein interacts with both OsTOP6A2 and OsTOP6A3, but not with OsTOP6A1. The nuclear localization of OsTOP6A3 and OsTOP6B was established by the transient expression of their beta-glucuronidase fusion proteins in onion epidermal cells. Overexpression of OsTOP6A3 and OsTOP6B in transgenic Arabidopsis plants conferred reduced sensitivity to the stress hormone, abscisic acid, and tolerance to high salinity and dehydration. Moreover, the stress tolerance coincided with enhanced induction of many stress-responsive genes in transgenic Arabidopsis plants. In addition, microarray analysis revealed that a large number of genes are expressed differentially in transgenic plants. Taken together, our results demonstrate that TOP6 genes play a crucial role in stress adaptation of plants by altering gene expression. PMID- 17116243 TI - Oxidation inhibits amyloid fibril formation of transthyretin. AB - The role of amino acid side chain oxidation in the formation of amyloid assemblies has been investigated. Chemical oxidation of amino acid side chains has been used as a facile method of introducing mutations on protein structures. Oxidation promotes changes within tertiary contacts that enable identification of residues and interactions critical in stabilizing protein structures. Transthyretin (TTR) is a soluble human plasma protein. The wild-type (WT) and several of its variants are prone to fibril formation, which leads to amyloidosis associated with many clinical syndromes. The effects of amino acid side chain oxidations were investigated by comparing the kinetics of fibril formation of oxidized and unoxidized proteins. The WT and V30M TTR mutant (valine 30 substituted with methionine) were allowed to react over a time range of 10 min to 12 h with hydroxy radical and other reactive oxygen species. In these timescales, up to five oxygen atoms were incorporated into WT and V30M TTR proteins. Oxidized proteins retained their tetrameric structures, as determined by cross-linking experiments. Side chain modification of methionine residues at position 13 and 30 (the latter for V30M TTR only) were dominant oxidative products. Mono-oxidized and dioxidized methionine residues were identified by radical probe mass spectometry employing a footprinting type approach. Oxidation inhibited the initial rates and extent of fibril formation for both the WT and V30M TTR proteins. In the case of WT TTR, oxidation inhibited fibril growth by approximately 76%, and for the V30M TTR by nearly 90%. These inhibiting effects of oxidation on fibril growth suggest that domains neighboring the methionine residues are critical in stabilizing the tetrameric and folded monomer structures. PMID- 17116244 TI - Huntingtin inclusion bodies are iron-dependent centers of oxidative events. AB - Recently, we reported that the transient expression of huntingtin exon1 polypeptide containing polyglutamine tracts of various sizes (httEx1-polyQ) in cell models of Huntington disease generated an oxidative stress whose intensity was CAG repeat expansion-dependent. Here, we have analyzed the intracellular localization of the oxidative events generated by the httEx1-polyQ polypeptides. Analysis of live COS-7 cells as well as neuronal SK-N-SH and PC12 cells incubated with hydroethidine or dichlorofluorescein diacetate revealed oxidation of these probes at the level of the inclusion bodies formed by httEx1-polyQ polypeptides. The intensity and frequency of the oxidative events among the inclusions were CAG repeat expansion-dependent. Electron microscopic analysis of cell sections revealed the presence of oxidation-dependent morphologic alterations in the vicinity of httEx1-polyQ inclusion bodies. Moreover, a high level of oxidized proteins was recovered in partially purified inclusions. We also report that the iron chelator deferroxamine altered the structure, localization and oxidative potential of httEx1-polyQ inclusion bodies. Hence, despite the fact that the formation of inclusion bodies may represent a defense reaction of the cell to eliminate httEx1 mutant polypeptide, this phenomenon appears inherent to the generation of iron-dependent oxidative events that can be deleterious to the cell. PMID- 17116245 TI - A ribonuclease zymogen activated by the NS3 protease of the hepatitis C virus. AB - Translating proteases as inactive precursors, or zymogens, protects cells from the potentially lethal action of unregulated proteolytic activity. Here, we impose this strategy on bovine pancreatic ribonuclease (RNase A) by creating a zymogen in which quiescent ribonucleolytic activity is activated by the NS3 protease of the hepatitis C virus. Connecting the N-terminus and C-terminus of RNase A with a 14-residue linker was found to diminish its ribonucleolytic activity by both occluding an RNA substrate and dislocating active-site residues, which are devices used by natural zymogens. After cleavage of the linker by the NS3 protease, the ribonucleolytic activity of the RNase A zymogen increased 105 fold. Both before and after activation, the RNase A zymogen displayed high conformational stability and evasion of the endogenous ribonuclease inhibitor protein of the mammalian cytosol. Thus, the creation of ribonuclease zymogens provides a means to control ribonucleolytic activity and has the potential to provide a new class of antiviral chemotherapeutic agents. PMID- 17116246 TI - Insulin-induced proliferation of bladder cancer cells is mediated through activation of the epidermal growth factor system. AB - The mechanism behind the growth-promoting effect of insulin is a subject of debate. Employing RT4 bladder cancer cells, we examined the cross-talk between insulin and the epidermal growth factor system. We found that insulin induced a time- and dose-dependent (25-1000 nmol.L(-1) insulin) increase in mRNA expression of three ligands from the epidermal growth factor system. Times for peak increase and fold increase after incubation with 250 nmol.L(-1) insulin were as follows: heparin-binding epidermal growth factor-like growth factor, 0.5 h, 1.4-fold, P < 0.05; epiregulin, 3 h, 14-fold, P < 0.0001; and amphiregulin, 3 h, 12-fold, P < 0.001. Induction of heparin-binding epidermal growth factor-like growth factor and amphiregulin was verified at the protein level. We demonstrate that incubation of RT4 bladder cancer cells for 24 h with 250 nmol.L(-1) insulin increases proliferation by 43% (P < 0.0001) as compared to untreated cells. At the same time, phosphorylation and thereby activation of the epidermal growth factor receptor (HER1) was observed. Both phosphorylation and insulin-induced proliferation were almost completely inhibited by the HER1 inhibitor Iressa (P < 0.0001). This shows that insulin leads to activation of HER1, and that HER1 plays an essential role in mediating the growth-promoting effect of insulin. Iressa inhibited not only the activation of HER1 caused by insulin but also the insulin induced increase in the three ligands (heparin-binding epidermal growth factor like growth factor, epiregulin and amphiregulin). As heparin-binding epidermal growth factor-like growth factor was induced before epiregulin and amphiregulin upon insulin stimulation, we speculated that the insulin-induced heparin-binding epidermal growth factor-like growth factor initiated the activation of HER1, and that this in turn led to increased expression of epiregulin and amphiregulin and thereby to continued activation of HER1. Earlier reports have shown that insulin like growth factor receptor can activate HER1 via its ligand heparin-binding epidermal growth factor-like growth factor. In accord with this, we found that treatment of RT4 cells with recombinant heparin-binding epidermal growth factor like growth factor mimicked the effect of insulin, with induction of mRNA for the three ligands. However, the insulin-induced increase in mRNA expression of amphiregulin and epiregulin could not be prevented by the heparin-binding epidermal growth factor-like growth factor inhibitor CRM197, demonstrating that heparin-binding epidermal growth factor-like growth factor is not essential for the insulin-induced increase in the expression of these ligands. In conclusion, we show that insulin-induced growth in RT4 cells requires activated HER1. Furthermore, activation of HER1 is required for the insulin-induced increase in expression of the HER1 ligands heparin-binding epidermal growth factor-like growth factor, amphiregulin and epiregulin. PMID- 17116247 TI - Intensive post-operative follow-up of breast cancer patients with tumour markers: CEA, TPA or CA15.3 vs MCA and MCA-CA15.3 vs CEA-TPA-CA15.3 panel in the early detection of distant metastases. AB - BACKGROUND: In breast cancer current guidelines do not recommend the routine use of serum tumour markers. Differently, we observed that CEA-TPA-CA15.3 (carcinoembryonic (CEA) tissue polypeptide (TPA) and cancer associated 115D8/DF3 (CA15.3) antigens) panel permits early detection and treatment for most relapsing patients. As high sensitivity and specificity and different cut-off values have been reported for mucin-like carcinoma associated antigen (MCA), we compared MCA with the above mentioned tumour markers and MCA-CA15.3 with the CEA-TPA-CA15.3 panel. METHODS: In 289 breast cancer patients submitted to an intensive post operative follow-up with tumour markers, we compared MCA (cut-off values, > or = 11 and > or = 15 U/mL) with CEA or CA15.3 or TPA for detection of relapse. In addition, we compared the MCA-CA15.3 and CEA-TPA-CA15.3 tumour marker panels. RESULTS: Distant metastases occurred 19 times in 18 (6.7%) of the 268 patients who were disease-free at the beginning of the study. MCA sensitivity with both cut-off values was higher than that of CEA or TPA or CA15.3 (68% vs 10%, 26%, 32% and 53% vs 16%, 42%, 32% respectively). With cut-off > or = 11 U/mL, MCA showed the lowest specificity (42%); with cut-off > or = 15 U/mL, MCA specificity was similar to TPA (73% vs 72%) and lower than that of CEA and CA15.3 (96% and 97% respectively). With > or = 15 U/mL MCA cut-off, MCA sensitivity increased from 53% to 58% after its association with CA15.3. Sensitivity of CEA-TPA-CA15.3 panel was 74% (14 of 19 recurrences). Eight of the 14 recurrences early detected with CEA-TPA-CA15.3 presented as a single lesion (oligometastatic disease) (5) or were confined to bony skeleton (3) (26% and 16% respectively of the 19 relapses). With > or = 11 U/mL MCA cut-off, MCA-CA15.3 association showed higher sensitivity but lower specificity, accuracy and positive predictive value than the CEA-TPA-CA15.3 panel. CONCLUSION: At both the evaluated cut-off values serum MCA sensitivity is higher than that of CEA, TPA or CA15.3 but its specificity is similar to or lower than that of TPA. Overall, CEA-TPA-CA15.3 panel is more accurate than MCA-CA15.3 association and can "early" detect a few relapsed patients with limited metastatic disease and more favourable prognosis. These findings further support the need for prospective randomised clinical trial to assess whether an intensive post-operative follow-up with an appropriate use of serum tumour markers can significantly improve clinical outcome of early detected relapsing patients. PMID- 17116248 TI - Temporal and spatial control of transgene expression using laser induction of the hsp70 promoter. AB - BACKGROUND: Precise temporal and spatial regulation of transgene expression is a critical tool to investigate gene function in developing organisms. The most commonly used technique to achieve tight control of transgene expression, however, requires the use of specific DNA enhancers that are difficult to characterize in non-model organisms. Here, we sought to eliminate the need for this type of sequence-based gene regulation and to open the field of functional genetics to a broader range of organisms. RESULTS: We have developed a new laser mediated method to heat shock groups of cells that provides precise spatio temporal control of gene expression without requiring knowledge of specific enhancer sequences. We tested our laser-system in a transgenic line of Bicyclus anynana butterflies containing the EGFP reporter gene attached to the heat sensitive hsp70 promoter of Drosophila melanogaster. Whole organismal heat shocks demonstrated that this Drosophila promoter can drive gene expression in butterflies, and the subsequent laser heat shocks showed that it was possible to activate cell-specific gene expression in very precise patterns on developing pupal wings. CONCLUSION: This laser-mediated gene expression system will enable functional genetic investigations, i.e., the ectopic expression of genes and their knock-down in targeted groups of cells in model and non-model organisms with little or no available regulatory data, as long as a compatible heat-shock promoter is used and the target tissue is accessible to a laser beam. This technique will also be useful in evolutionary developmental biology as it will enable the study of the evolution of gene function across a variety of organisms. PMID- 17116249 TI - CoXpress: differential co-expression in gene expression data. AB - BACKGROUND: Traditional methods of analysing gene expression data often include a statistical test to find differentially expressed genes, or use of a clustering algorithm to find groups of genes that behave similarly across a dataset. However, these methods may miss groups of genes which form differential co expression patterns under different subsets of experimental conditions. Here we describe coXpress, an R package that allows researchers to identify groups of genes that are differentially co-expressed. RESULTS: We have developed coXpress as a means of identifying groups of genes that are differentially co-expressed. The utility of coXpress is demonstrated using two publicly available microarray datasets. Our software identifies several groups of genes that are highly correlated under one set of biologically related experiments, but which show little or no correlation in a second set of experiments. The software uses a re sampling method to calculate a p-value for each group, and provides several methods for the visualisation of differentially co-expressed genes. CONCLUSION: coXpress can be used to find groups of genes that display differential co expression patterns in microarray datasets. PMID- 17116250 TI - Early home-based recognition of anaemia via general danger signs, in young children, in a malaria endemic community in north-east Tanzania. AB - BACKGROUND: Ethnographic studies from East Africa suggest that cerebral malaria and anaemia are not classified in local knowledge as malaria complications, but as illnesses in their own right. Cerebral malaria 'degedege' has been most researched, in spite of anaemia being a much more frequent complication in infants, and not much is known on how this is interpreted by caretakers. Anaemia is difficult to recognize clinically, even by health workers. METHODS: Ethnographic longitudinal cohort field study for 14 months, with monthly home visits in families of 63 newborn babies, identified by community census, followed throughout April - November 2003 and during follow-up in April-May 2004. Interviews with care-takers (mostly mothers) and observational studies of infants and social environment were combined with three haemoglobin (Hb) screenings, supplemented with reports from mothers after health facility use. RESULTS: General danger signs, reported by mothers, e.g. infant unable to breast-feed or sit, too weak to be carried on back - besides of more alarming signs such as sleeping all time, loosing consciousness or convulsing - were well associated with actual or evolving moderate to severe anaemia (Hb 75 (group E) and from 8 younger patients aged 50-60 (considered as controls, group C) were used for immunohistochemistry with monoclonal antibodies against CD45RB (leucocytes), CD1a (LC), markers of LC maturation (DC-LAMP, CD83) and number of immunolabelled cell subsets was evaluated using image analysis. RESULTS: The difference in the number of CD45RB+ leucocytes in the upper connective tissue between groups was not significant. In group E, the number of CD1a+ LC was significantly decreased (P<0.002) in the epithelium and significantly increased (P<0.0004) in the upper connective tissue. Furthermore, in group E, intraepithelial CD1a+ LC are more often observed in the upper epithelium and their dendritic processes were shorter and less numerous. Concerning the expression of markers of maturation, the numbers of intraepithelial DC-LAMP+ cells and CD83+ cells were significantly increased (P<0.0007 and P<0.02, respectively) in group E. CONCLUSION: During chronic periodontitis in elderly patients, the decrease in the number of intraepithelial LC and the alteration of dendritic processes could be balanced by a cellular distribution often observed in the upper epithelium associated with changes in cell maturation in response to bacterial elements. PMID- 17116294 TI - Dietary n-3 fatty acids promote arrhythmias during acute regional myocardial ischemia in isolated pig hearts. AB - OBJECTIVE: Dietary supplementation with fish oil-derived n-3 fatty acids reduces mortality in patients with myocardial infarction, but may have adverse effects in angina patients. The underlying electrophysiologic mechanisms are poorly understood. We studied the arrhythmias and the electrophysiologic changes during regional ischemia in hearts from pigs fed a diet rich in fish oil. METHODS: Pigs received diets rich in fish oil, in sunflower oil, or a control diet for 8 weeks. Hearts were isolated and perfused. Ischemia was created by occluding the left anterior descending artery. Diastolic stimulation threshold, refractory period, conduction velocity, activation recovery intervals and the maximum downstroke velocity of 176 electrograms were measured in the ischemic zone. Spontaneous arrhythmias during 75 min of regional ischemia were counted. RESULTS: More episodes of spontaneous ischemia-induced sustained ventricular tachycardia and ventricular fibrillation occurred in the fish oil and sunflower oil group than in the control group. More inexcitable myocardium was present in the ischemic zone in the group fed fish oil or sunflower oil than in the control group after 20 min of ischemia. After 40 min of ischemia, more block occurred in the control group than in the other groups. The downstroke velocity of the electrograms in the ischemic border zone was lower in the fish oil group and sunflower oil group than in the control after 20 min. CONCLUSIONS: A diet rich in fish oil results in proarrhythmia compared to a control diet during regional ischemia in pigs. Myocardial excitability is reduced in the fish oil and sunflower oil group during the early phase of arrhythmogenesis. In the late phase of arrhythmogenesis, excitability is more reduced in the control group than in the fish oil and sunflower oil group. PMID- 17116295 TI - A new ATP-sensitive potassium channel opener protects endothelial function in cultured aortic endothelial cells. AB - OBJECTIVE: Endothelial dysfunction is an early risk factor for cardiovascular disease and hypertension. Mechanisms that participate in endothelial dysfunction include reduced nitric oxide (NO) generation and increased endothelin-1 (ET-1) generation. Endothelial ATP-sensitive potassium (K(ATP)) channels are responsible for maintaining the resting potential of endothelial cells and modulating the release of vasoactive compounds. We hypothesized that activation of endothelial K(ATP) channels might result in the protection against endothelial dysfunction. METHODS: Using cultured bovine or rat aortic endothelial cells, we examined the effects of a new K(ATP) channels opener, iptakalim, on the secretion of vasoactive substances. We also investigated its effects on the expression of adhesion molecules in metabolically disturbed cultured endothelial cells. RESULTS: In cultured aortic endothelial cells, iptakalim caused a concentration dependent inhibition of ET-1 release and synthesis that correlated with reduced levels of mRNA for ET-1 and endothelin-converting enzyme. These effects of iptakalim were significantly inhibited by pretreatment with glibenclamide (a K(ATP) channel blocker) for 1 h. Similarly, iptakalim enhanced the release of NO in a concentration-dependent manner and increased basal levels of free intracellular calcium. Iptakalim at the concentrations of 100 and 1000 microM increased the activities of NO synthase (NOS) significantly. After the activity of NOS was blocked by L-N(omega)-nitro-arginine methyl ester (L-NAME), the inhibition of iptakalim on ET-1 release was abolished. In endothelial cell models of metabolic disturbance induced by low-density lipoprotein, homocysteine, or hyperglycemia, treatment with iptakalim could inhibit the overexpression of monocyte chemoattractant protein-1 (MCP-1), Intercellular adhesive molecule-1 (ICAM-1), and vascular cell adhesive molecule-1 (VCAM-1) mRNA. CONCLUSION: Iptakalim is a promising drug that could protect against endothelial dysfunction through activating K(ATP) channels in endothelial cells. PMID- 17116297 TI - Calorimetric study on the induction of interdigitated phase in hydrated DPPC bilayers by bioactive labdanes and correlation to their liposome stability: The role of chemical structure. AB - Labd-7,13-dien-15-ol (1), labd-13-ene-8alpha,15-diol (2), and labd-14-ene-8,13 diol (sclareol) have been found to exhibit cytotoxic and cytostatic effects. Their partitioning into phospholipid bilayers may induce membrane structure modifications, crucial in the development of liposomes. DSC was used to elucidate the profile of modifications induced in DPPC bilayers by incorporating increasing concentrations of the labdanes. Labdanes 1, 2 and sclareol were incorporated into SUV liposomes composed of DPPC their physicochemical stability was monitored (4 degrees C) and was compared to liposomes incorporating cholesterol. All labdanes strongly affect the bilayer organization in a concentration dependent manner in terms of a decrease of the cooperativity, the fluidization and partially destabilization of the gel phase, the induction of a lateral phase separation and the possible existence of interdigitated domains in the bilayer. The physicochemical stability of liposomes was strongly influenced by the chemical features of the labdanes. The liposomal preparations were found to retain their stability at low labdane concentration (10 mol%), while at higher concentrations up to 30 mol% a profound decrease in intact liposomes occurred, and a possible existence of interdigitated sheets was concluded. PMID- 17116296 TI - The protective efficacy of chlamydial protease-like activity factor vaccination is dependent upon CD4+ T cells. AB - We have previously determined the protective efficacy of intranasal vaccination with chlamydial protease-like activity factor (CPAF) against genital chlamydial infection. Since T-helper 1 (Th1) responses are important for anti-chlamydial immunity, we examined the contribution of CD4(+) T cells in CPAF mediated immunity against intravaginal (i.vag.) Chlamydia muridarum infection in C57BL/6 mice. CPAF+IL-12 vaccination induced antigen-specific CD4(+) T cells that secreted elevated levels of IFN-gamma, and generated strong humoral responses. The protective effects of CPAF vaccination against genital chlamydial challenge were abrogated by anti-CD4 neutralizing antibody treatment. Moreover, anti chlamydial immunity could be adoptively transferred to naive recipients using CPAF-specific CD4(+) T cells. Therefore, CPAF mediated anti-chlamydial immunity is highly dependent upon antigen-specific CD4(+) T cells. PMID- 17116298 TI - Applying proteomic methodologies to analyze the effect of methionine restriction on proliferation of human gastric cancer SGC7901 cells. AB - BACKGROUND: Methionine dependence is a feature unique to cancer cells, exhibited as inability to grow in a methionine-depleted environment supplemented with homocysteine, the immediate metabolic precursor of methionine. However, the molecular mechanisms by which methionine restriction inhibits cancer cells growth have not been elucidated. The effect of methionine restriction on the protein expression in gastric cancer cells was studied. METHODS: SGC7901 cells were treated with M-H+ medium for 5 days, which was followed by analysis of total cellular protein from cells by a combination of 2-DE and MS. Then the differential expressional levels of partially identified proteins were determined by Western blot analysis. RESULTS: The well-resolved, reproducible 2-DE patterns of SGC7901 cells cultured in M+H- or M-H+ medium were established. The 10 differential proteins between pairs of gastric cancer cells SGC7901 cultured either in M+H- medium or M-H+ medium, were identified by MALDI-TOF/TOF MS, and the differential expression levels of 2 identified proteins were confirmed. CONCLUSION: These data will be valuable for further study of the molecular mechanisms by which methionine restriction induces cell cycle arrest and apoptosis in human gastric cancer. PMID- 17116299 TI - 5-HT1A receptors are involved in the anxiolytic effect of Delta9 tetrahydrocannabinol and AM 404, the anandamide transport inhibitor, in Sprague Dawley rats. AB - The mechanism mediating the effects of cannabinoids on anxiety-related responses appear to involve cannabinoid CB1 and non-CB1 receptors. However, other neurotransmitters may play a role in such effect. This study shows evidence of an interaction between endocannabinoid system and serotonin (5-HT), 1A receptor subtype on anxiety-like behavior in Sprague-Dawley rats. The exogenous cannabinoid agonist, Delta9-tetrahydrocannabinol (THC), and N-(4-hydroxyphenyl) arachidonylamide, the anandamide transporter inhibitor (AM 404) were evaluated in the elevated plus maze test. THC (0.075-0.75 mg/kg i.p.), given 30 min and AM 404 (0.75-1.25 mg/kg i. p.), given 60 min before the test, exhibited a dose-response anxiolytic effect evaluated in terms of increase in the percentage of total entries and time spent in the open and decrease of total entries and time spent in the closed arms. The anxiolytic effect obtained with the maximal active dose of both THC (0.75 mg/kg) and AM 404 (1.25 mg/kg) was blocked by the 5-HT1A receptor antagonist, N-[2-[4-(2-methoxyphenyl) piperazin-1-yl]ethyl]-N-pyridin-2 yl-cyclohexanecarboxamide dihydro chloride (WAY-100635 (300 microg/kg, s.c.), given 30 min before THC or 15 min before AM 404. The combination of an ineffective dose of THC (0.015 mg/kg) or AM 404 (0.015 mg/kg) on anxiety-related responses with an ineffective dose of the 5HT(1A) receptor agonist, 8-Hydroxy-2 (di-n-propylamino) tetralin hydrobromide (8-OH-DPAT) (7.5 microg/kg, i.p.), led to a synergistic effect. No interference with spontaneous motor activity, evaluated in an activity cage for 5 min, in rats given the drugs alone or in combination, was found. These data suggest that the anxiolytic effect produced by endo- and eso-cannabinoids is modulated by 5-HT1A receptors. PMID- 17116300 TI - The mechanism of apoptosis induced by a novel thioredoxin reductase inhibitor in A549 cells: possible involvement of nuclear factor-kappaB-dependent pathway. AB - 1,2-[bis(1,2-benzisoselenazolone-3(2H)-ketone)]ethane (BBSKE, PCT: CN02/00412), a novel thioredoxin reductase inhibitor previously synthesized in our lab, has been demonstrated to inhibit the growth of a variety of human cancer cells and to induce apoptosis. Here we report on the potential molecular mechanism of apoptosis induced by BBSKE in A549 cells. The treatment of BBSKE reduced the protein levels of Bcl-2, Bcl-xL, procaspase-9 and procaspase-3, and caused the release of cytochrome C from the mitochondria to the cytosol in a dose-dependent manner, suggesting the onset of mitochondria-dependent apoptosis. Through electrophoretic mobility shift assay (EMSA), the DNA-binding activity of nuclear factor-kappaB (NF-kappaB) was found to be attenuated after BBSKE treatment, accompanied by the diminution of the immunoprecipitated complex of thioredoxin and NF-kappaB in co-immunoprecipitation experiments. Meanwhile, the ratio of pIkappaB-alpha to IkappaB-alpha and the subcellular localization of p65 between cytoplasm and nucleus were not significantly altered by BBSKE treatment, as demonstrated in western analysis and immunocytochemistry assay. Furthermore, the mRNA levels of the NF-kappaB regulated anti-apoptosis genes Bcl-2, Bcl-xL, cIAP-2 and XIAP were decreased in a dose-dependent manner after BBSKE treatment. All the above observations suggest that BBSKE induce mitochondria-dependent apoptosis in A549 cells probably through suppressing the thioredoxin reductase-thioredoxin-NF kappaB pathway. PMID- 17116301 TI - Antidepressant-induced undesirable weight gain: prevention with rimonabant without interference with behavioral effectiveness. AB - Antidepressant pharmacotherapy has dramatically improved the quality of life for many patients. However, prolonged use may induce weight gain, resulting in enhanced risk for treatment noncompliance. Cannabinoid CB(1) receptor antagonists decrease food intake and body weight, but may also affect mood. We investigated in female Sabra mice first, whether acute treatment with the cannabinoid receptor antagonist rimonabant (5-(4-Chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-N (piperidin-1-yl)-1H-pyrazole-3-carboxamide, SR141716, 5 mg/kg) interfered with the tricyclic antidepressant desipramine (15 mg/kg) or the selective serotonin reuptake inhibitor fluoxetine (20 mg/kg) in the Porsolt forced swimming test. Second, whether chronic treatment (3 months) with desipramine (5 mg/kg) enhanced weight gain and whether cotreatment with rimonabant (2 mg/kg), prevented the excessive weight gain, while retaining antidepressant effectiveness. Motor activity and anxiety-like behavior were also investigated. The acute studies indicated that rimonabant did not influence 'antidepressant' activity of desipramine or fluoxetine. In the chronic studies, desipramine enhanced weight gain, despite the observation that the injection procedure reduced weight gain. The enhanced weight gain continued at least 35 days after treatment ended. Rimonabant reduced weight gain to which no tolerance developed and which persisted at least 30 days beyond treatment. Mice cotreated with rimonabant and desipramine had body weights closer to controls or to those receiving rimonabant alone than to those treated with desipramine alone. The antidepressant effects of desipramine were maintained throughout treatment; this was not altered by the chronic rimonabant treatment at any time, although rimonabant together with desipramine transiently enhanced anxiety-like behavior. These observations suggest that combined treatment with antidepressants and cannabinoid CB(1) receptor antagonist to prevent undesirable weight gain, should be further investigated. PMID- 17116302 TI - Cellular pharmacological properties of gold(III) porphyrin 1a, a potential anticancer drug lead. AB - The development of gold(III) complexes as potential anticancer drugs with higher cytotoxicity and fewer side effects than existing metal anticancer drugs has been actively pursued in recent years. In this study, we explored the cellular pharmacological properties of gold(III) porphyrin 1a, an anticancer drug lead we previously described. The cytotoxicity study of gold(III) porphyrin 1a by naphthol blue black (NBB) staining assay demonstrated that the higher cytotoxicity of gold(III) porphyrin 1a was not related to its photosensitizing activity. Serum dependent test revealed that serum proteins exhibited lesser effects on the activity of gold(III) porphyrin 1a. In addition, in vivo and in vitro binding assays showed that gold(III) porphyrin 1a acted on DNA noncovalently, which was differently from cisplatin. Flow cytometric study indicated that gold(III) porphyrin 1a inhibited cell growth partly through abrogating cell cycle at G(0)-G(1), and induced apoptosis in SUNE1 cells. The enhanced expression of p53, a cell cycle-controlling and apoptosis-related protein, further demonstrated that the cell cycle arrest and apoptosis induced by gold porphyrin 1a were p53 dependent. Our results highlighted the potential of gold(III) porphyrin 1a as an anticancer drug. PMID- 17116303 TI - Dispersion-convolution model for simulating peaks in a flow injection system. AB - A dispersion-convolution model is proposed for simulating peak shapes in a single line flow injection system. It is based on the assumption that an injected sample plug is expanded due to a "bulk" dispersion mechanism along the length coordinate, and that after traveling over a distance or a period of time, the sample zone will develop into a Gaussian-like distribution. This spatial pattern is further transformed to a temporal coordinate by a convolution process, and finally a temporal peak image is generated. The feasibility of the proposed model has been examined by experiments with various coil lengths, sample sizes and pumping rates. An empirical dispersion coefficient (D*) can be estimated by using the observed peak position, height and area (tp*, h* and At*) from a recorder. An empirical temporal shift (Phi*) can be further approximated by Phi*=D*/u2, which becomes an important parameter in the restoration of experimental peaks. Also, the dispersion coefficient can be expressed as a second-order polynomial function of the pumping rate Q, for which D*(Q)=delta0+delta1Q+delta2Q2. The optimal dispersion occurs at a pumping rate of Qopt=sqrt[delta0/delta2]. This explains the interesting "Nike-swoosh" relationship between the peak height and pumping rate. The excellent coherence of theoretical and experimental peak shapes confirms that the temporal distortion effect is the dominating reason to explain the peak asymmetry in flow injection analysis. PMID- 17116304 TI - Hydrophobic interaction chromatography of proteins IV. Kinetics of protein spreading. AB - Adsorption of proteins on surfaces of hydrophobic interaction chromatography media is at least a two-stage process. Application of pure protein pulses (bovine serum albumin and beta-lactoglobulin) to hydrophobic interaction chromatography media yielded two chromatographic peaks at low salt concentrations. At these salt concentrations, the adsorption process is affected by a second reaction, which can be interpreted as protein spreading or partial unfolding of the protein. The kinetic constants of the spreading reaction were derived from pulse response experiments at different residence times and varying concentrations by applying a modified adsorption model considering conformational changes. The obtained parameters were used to calculate uptake and breakthrough curves for spreading proteins. Although these parameters were determined at low saturation of the column, predictions of overloaded situations could match the experimental runs satisfactorily. Our findings suggest that proteins which are sensitive to conformational changes should be loaded at high salt concentrations in order to accelerate the adsorption reaction and to obtain steeper breakthrough curves. PMID- 17116305 TI - Automation and optimization of liquid-phase microextraction by gas chromatography. AB - Several fully automated liquid-phase microextraction (LPME) techniques, including static headspace LPME (HS-LPME) (a drop of solvent is suspended at the tip of a microsyringe needle and exposed to the headspace of the sample solution), exposed dynamic HS-LPME (the solvent is exposed in the headspace of sample vial for different time, and then withdrawn into the barrel of the syringe. This procedure is repeated a number of times), unexposed dynamic HS-LPME (the solvent is moved inside the needle and the barrel of a syringe, and the gaseous sample is withdrawn into the barrel and then ejected), static direct-immersed LPME (DI LPME) (a drop of solvent is suspended at the tip of a microsyringe needle and directly immersed into the sample solution), dynamic DI-LPME (the solvent is moved inside the needle and the barrel of a syringe, and the sample solution is withdrawn and ejected), and two phase hollow fiber-protected LPME (HF-LPME) (a hollow fiber is used to stabilize and protect the solvent), auto-performed with a commercial CTC CombiPal autosampler, are described in this paper. Critical experimental factors, including temperature, choice of extraction solvent, solvent volume, plunger movement rate, and extraction time were investigated. Among the three HS-LPME techniques that were evaluated, the exposed dynamic HS LPME technique provided the best performance, compared to the unexposed dynamic HS-LPME and static HS-LPME approaches. For DI-LPME, the dynamic process can enhance the extraction efficiency and the achieved method precision is comparable with the static DI-LPME technique. The precision of the fully automated HF-LPME is quite acceptable (RSD values below 6.8%), and the concentration enrichment factors are better than the DI-LPME approaches. The fully automated LPME techniques are more accurate and more convenient, and the reproducibility achieved eliminates the need for an internal standard to improve the method precision. PMID- 17116306 TI - Study of dynamic adsorption behavior of large-size protein-bearing particles. AB - The subject of this paper is an investigation of the peculiarities of dynamic adsorption behavior of nanoparticles. For this purpose, virus-mimicking synthetic particles bearing different proteins at their outer surface were specially constructed using two approaches, e.g. the cross-linking of proteins and modification of polystyrene microsphere surface by proteins. Two chromatographic modes, namely ion-exchange and affinity liquid chromatography on ultra-short monolithic columns [Convective Interaction Media (CIM) DEAE and CIM QA disks] have been used as a tool for dynamic adsorption experiments. Such parameters as maximum adsorption capacity and its dependence on applied flow rate were established and compared with those obtained for individual proteins. Similarly to individual proteins, it was shown that the maximum of adsorption capacity was not changed at different flow rates. In addition, the permeability of porous space of used monolithic sorbents appeared to be sufficient for efficient separation of large particles and quite similar to the well-studied process applied for individual proteins. PMID- 17116307 TI - Glass-fiber reinforced poly(acrylate)-based sorptive materials for the enrichment of organic micropollutants from aqueous samples. AB - A novel and simple approach to sorptive materials for the extraction of organic compounds from environmental samples is presented. It entails the use of glass fiber fabric strips coated with a customized poly(acrylate) (PA) formulation as extraction medium. Analytes were enriched by means of shaking of the PA strip in the sample and then subsequently thermally desorbed and analyzed by GC-MS. The performance of the sorptive materials was evaluated by the enrichment of compounds with different polarities (phenols, hexachlorobenzene and hexachlorocyclohexanes) from water samples. Parameters that affect the extraction process such as pH, ionic strength of the solvent, mixing mechanism, extraction time and desorption conditions were investigated. The extraction abilities of the PA extraction medium were compared with that of commercially available poly(dimethylsiloxane) (PDMS) Twisters. The results revealed that the new material shows higher affinity for phenolic compounds presumably due to the presence of polar groups. The partition coefficients for the PA strips estimated in this work were up to 15 times higher than for the PDMS Twister, resulting in higher recoveries and lower detection limits. The method was applied for the quantification of the aforementioned compounds in contaminated surface water samples from Bitterfeld (Germany). Using both PA strips and PDMS Twisters, good agreement of the extraction data was achieved. PMID- 17116308 TI - Ligand-regulated peptide aptamers that inhibit the 5'-AMP-activated protein kinase. AB - In an effort to extend the peptide aptamer approach, we have developed a scaffold protein that allows small molecule ligand control over the presentation of a peptide aptamer. This scaffold, a fusion of three protein domains, FKBP12, FRB, and GST, presents a peptide linker region for target protein binding only in the absence of the small molecule Rapamycin or other non-immunosuppressive Rapamycin derivatives. Here we describe the characterization of ligand-regulated peptide aptamers that interact with and inhibit the 5'-AMP-activated protein kinase (AMPK). AMPK, a central regulator of cellular energy homeostasis, responds to high cellular AMP/ATP ratios by promoting energy producing pathways and inhibiting energy consuming biosynthetic pathways. We have characterized 15 LiRPs of similar, poly-basic sequence and have determined that they interact with the substrate peptide binding region of both AMPK alpha1 and alpha2. These proteins, some of which serve as poor substrates of AMPK, inhibit the kinase as pseudosubstrates in a Rapamycin-regulated fashion in vitro, an effect that is largely competitive with substrate peptide and mediated by an increase in the kinase's apparent K(m) for substrate peptide. This pseudosubstrate inhibition of AMPK by LiRP proteins reduced the AMP stimulation of AMPK in vitro and caused the inhibited state of the kinase to kinetically resemble the basal, unstimulated state of AMPK, providing potential insight into the molecular mechanisms of AMP stimulation of AMPK. PMID- 17116309 TI - Tamoxifen effect on L-DOPA induced response complications in parkinsonian rats and primates. AB - The contribution of striatal protein kinase C (PKC) isoform changes in levodopa (L-DOPA) induced motor response complications in parkinsonian rats was investigated and the ability of tamoxifen, an antiestrogen with a partial PKC antagonist property, to prevent these response alterations in 6-hydroxydopamine (6-OHDA) lesioned rats as well as in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treated cynomologous monkeys was studied. Following treatment of adult male rats with L-DOPA twice daily for 3 weeks, protein levels of left (lesioned) and right (intact) striatal PKC isoforms were measured. Western blot analysis showed increased protein expression of both the novel PKC epsilon isoform and the atypical PKC lambda isoform ipsilateral to the lesion (174+/-17% for epsilon, 140+/-9% for lambda, of intact striatum in 6-OHDA lesioned plus chronic L-DOPA treated animals) in acute L-DOPA treated rats. No enhancement was observed in PKC immunoreactivity for other isoforms. Tamoxifen (5.0 mg/kg p.o.) significantly attenuated the L-DOPA induced augmentation of protein expression of PKC epsilon and PKC lambda, but had no effect on immunoreactivity for other PKC isoforms. In chronic L-DOPA treated parkinsonian rats, tamoxifen prevented (5.0 mg/kg p.o.) as well as ameliorated (5.0 mg/kg p.o.) the characteristic shortening in duration of motor response to L-DOPA challenge. In MPTP lesioned primates, similar to the ameliorative effect seen in rats, tamoxifen (1 and 3 mg/kg p.o) reduced the appearance of L-DOPA induced dyskinesia by 61% and 55% respectively (p<0.05). These results suggest that changes in specific striatal PKC isoforms contribute to the pathogenesis of L-DOPA induced motor complications and further that drugs able to selectively inhibit these signaling kinases might provide adjunctive benefit in the treatment of Parkinson's disease. PMID- 17116310 TI - Double dissociating effects of sensory stimulation and cocaine on serotonin activity in the occipital and temporal cortices. AB - Visual cues that become associated with the consumption of psychostimulant drugs energize craving and the intake of the drug by mechanisms of which little is known. In two experiments using in vivo microdialysis in freely moving rats we compared the effects of visual and auditory stimulation with that of cocaine (0, 5, 10, 20mg/kg; i.p.) on the extracellular serotonin (5-HT) activity in the occipital and temporal cortices in relation to behavior. Visual stimulation increased 5-HT in the occipital, but not temporal cortex, parallel to an increase in locomotion. Auditory stimulation decreased 5-HT in the auditory, but not occipital cortex, thus, showing a double dissociated 5-HT response. These data suggest that a locally restricted 5-HT response to sensory stimulation may gate behavioral activity sense-modality selectively. Cocaine affected 5-HT in the occipital cortex and behavioral activity in the same direction as visual stimulation, but in an amplified and prolonged way. In the temporal cortex cocaine also caused an increase in 5-HT. The findings demonstrate common effects of visual stimulation and cocaine on 5-HT activity in the occipital cortex in relation to locomotor activity. The results suggest that concepts of how neutral visual cues become powerful energizers of addiction-related behaviors should be expanded to incorporate not only an acute enhancement of reward processing mechanisms, but, in parallel, also an amplified processing of visual stimuli in the occipital cortex. PMID- 17116311 TI - Pattern recognition using a device substituting audition for vision in blindfolded sighted subjects. AB - A major question in the field of sensory substitution concerns the nature of the perception generated by sensory substitution devices. In the present fMRI study, we investigated the neural substrates of pattern recognition through a device substituting audition for vision in blindfolded sighted subjects, before and after a short training period. Before training, pattern recognition recruited dorsal and ventral extra-striate areas. After training, the recruitment of these visual areas was found to have increased. These results suggest that visual imagery processes could be involved in pattern recognition and that perception through the substitution device could be visual-like. PMID- 17116312 TI - Enhanced circadian photoresponsiveness after prolonged dark adaptation in seven species of diurnal and nocturnal rodents. AB - Previous studies in mice and Syrian hamsters have described an enhancement of circadian photoresponsiveness after exposure to darkness for several weeks. The present study investigated the generality of the phenomenon in 3 diurnal and 4 nocturnal rodent species. In four of the species tested, phase delays of the running-wheel activity rhythm evoked by 1-h light pulses were several-fold larger after 3 to 4 weeks of exposure to darkness than after a single day. This drastic change in photoresponsiveness has important implications for the understanding of the process of photic entrainment. Differences between species that showed a significant effect of dark adaptation and species that showed no effect were not accounted for by temporal niche (diurnal versus nocturnal) or photic sensitivity (albino versus pigmented). Further research is needed to elucidate the mechanisms responsible for inter-species differences in the occurrence of enhanced photoresponsiveness after dark adaptation and to identify the neural substrates of this phenomenon in species that exhibit it. PMID- 17116313 TI - D-homoannulation of 17alpha,21-dihydroxy-20-keto steroids (corticosteroids). AB - Synthetic corticosteroids are widely used as anti-inflammatory agents. Mechanisms of their degradation continue to be studied. D-ring homoannulation is a well known metabolic pathway for steroids in vivo. The rearrangement with aluminium trichloride of the commercial anti-inflammatory drugs hydrocortisone, cortisone and dexamethasone is here presented. The structures of the corresponding 17a-keto 17-hydroxy-D-homosteroids are established by mono- and two-dimensional NMR analysis. Inversion of the alpha-configuration of C-16 is observed in the Lewis acid assisted D-homoannulation of dexamethasone. PMID- 17116314 TI - Attention makes moving objects be perceived to move faster. AB - Although it is well established that attention affects visual performance in many ways, by using a novel paradigm [Carrasco, M., Ling, S., & Read. S. (2004). Attention alters appearance. Nature Neuroscience, 7, 308-313.] it has recently been shown that attention can alter the perception of different properties of stationary stimuli (e.g., contrast, spatial frequency, gap size). However, it is not clear whether attention can also change the phenomenological appearance of moving stimuli, as to date psychophysical and neuro-imaging studies have specifically shown that attention affects the adaptability of the visual motion system. Here, in five experiments we demonstrated that attention effectively alters the perceived speed of moving stimuli, so that attended stimuli were judged as moving faster than less attended stimuli. However, our results suggest that this change in visual performance was not accompanied by a corresponding change in the phenomenological appearance of the speed of the moving stimulus. PMID- 17116315 TI - Accumulation of senescent cells in mitotic tissue of aging primates. AB - Cellular senescence, a stress induced growth arrest of somatic cells, was first documented in cell cultures over 40 years ago, however its physiological significance has only recently been demonstrated. Using novel biomarkers of cellular senescence we examined whether senescent cells accumulate in tissues from baboons of ages encompassing the entire lifespan of this species. We show that dermal fibroblasts, displaying markers of senescence such as telomere damage, active checkpoint kinase ATM, high levels of heterochromatin proteins and elevated levels of p16, accumulate in skin biopsies from baboons with advancing age. The number of dermal fibroblasts containing damaged telomeres reaches a value of over 15% of total fibroblasts, whereas 80% of cells contain high levels of the heterochromatin protein HIRA. In skeletal muscle, a postmitotic tissue, only a small percentage of myonuclei containing damaged telomeres were detected regardless of animal age. The presence of senescent cells in mitotic tissues might therefore be a contributing factor to aging and age related pathology and provides further evidence that cellular senescence is a physiological event. PMID- 17116316 TI - The genetics and epigenetics of altered proliferative homeostasis in ageing and cancer. AB - Ageing mammals are subject to an amazing array of aberrations in proliferative homeostasis. These are of two basic types: the post-maturational failure to adequately replace effete somatic cells (atrophies) and excessive proliferations of somatic cells (hyperplasias). To a surprising degree, these occur side by side within the same tissues and are features of numerous mammalian geriatric disorders. Atrophy is the likely usual initial event, the proliferative response perhaps developing as a secondary, compensatory, initially adaptive reaction. We have little understanding of why this putative compensatory reaction so often fails to be appropriately regulated in ageing mammals, leading to such pathologies as chronic inflammation, fibrosis, metaplasia and neoplasia. Advances in formal genetic analysis, mutagenesis, stem cell biology and epigenetics are likely to provide major new understanding. Stochastic epigenetic shifts in gene expression are of growing interest, particularly in explaining intra-specific variations on rates and patterns of ageing. Nature may well have evolved such random fluctuations in gene expression as a type of group-selectionist adaptive strategy to cope with diverse stochastic environmental challenges. Alternatively, such background "noise" in transcription and translation may simply reflect a type of informational entropy. PMID- 17116317 TI - Susceptibility groups for Alzheimer's disease (OPTIMA cohort): integration of gene variants and biochemical factors. AB - Information on gene variants and blood levels (APOE, BCHE-K, TF-C2, HFE-D, HFE-Y, ACE I/D, AR1; homocysteine, folate and vitamin B(12)) is available for participants in the Oxford Project to Investigate Memory and Ageing (OPTIMA) cohort (n=575). This information identified four risk sets for Alzheimer's disease (AD) using grade of membership analysis (GoM). Graded membership scores that relate individuals to each set are automatically generated. Sets I and III had low intrinsic risk. Set II had high intrinsic risk associated with multiple gene variants, e.g., APOE44/34. Set IV also had high intrinsic risk demonstrating low folate and B(12) levels. Membership in the high intrinsic risk sets was summed, coded as either close versus not close (>or=0.80 versus <0.80) and input into logistic models to predict relative risk: close resemblance multiplied risk 80-fold for possible AD before age 65 and 55-fold for probable or definite AD at ages 65-74. These findings implicate both biochemical and genetic factors in the risk for AD and further support dietary supplementation with folate and vitamin B(12) as a potential means to delay the onset of AD and/or its rate of progression. PMID- 17116318 TI - Is cell death and replacement a factor in aging? AB - The central theme of the 3rd International Conference on Functional Genomics of Ageing was tissue regeneration as a remedial strategy to address age-related cellular damage and the pathology that ensues. The conference included sessions on maintaining genome integrity and the potential of stem cells to restore function to damaged tissues. In addition to several human syndromes that appear to reflect accelerated ageing, there are now a number of mouse models that prematurely display phenotypes associated with ageing. The intent of this summary presented at the end of the conference was to: (1) discuss various human syndromes and mouse models of accelerated ageing; (2) evaluate whether the phenotypes displayed might result from an elevated rate of cell death coupled with an inability to adequately maintain cell number in various tissues with increasing age; and (3) discuss whether similar events may be occurring during normal ageing, albeit much more slowly. PMID- 17116319 TI - Polymorphisms of pro-inflammatory genes and Alzheimer's disease risk: a pharmacogenomic approach. AB - Clinically and pathologically Alzheimer's disease (AD) represents a sequential progressive neurodegenerative disorder. AD is etiologically heterogeneous and accounts for a majority of dementia in western societies. Inflammation clearly occurs in pathologically vulnerable regions of the AD brain and the search for genetic factors influencing the pathogenesis of AD has lead to the identification of numerous gene polymorphisms that might act as susceptibility modifiers. Accordingly, several reports have indicated that the risk of AD is substantially influenced by several genetic polymorphisms in the promoter region, or other untranslated regions, of genes encoding inflammatory mediators, although not all the studies were replied. Here, we review several data suggesting that inflammatory genetic variation may contribute to AD susceptibility. All together this information may represent the basis both for future recognition of individuals at risk and for the pharmacogenomic driving of drug responsiveness. PMID- 17116320 TI - The peroxynitrite decomposition catalyst FP15 improves ageing-associated cardiac and vascular dysfunction. AB - Overproduction of oxidants and free radicals in ageing tissues induces nitro oxidative stress, which has recently been implicated in the pathogenesis of cardiovascular dysfunction associated with ageing. Peroxynitrite, a strong cytotoxic oxidant damages proteins and DNA and activates several pathways causing tissue injury, including the peroxynitrite-poly(ADP-ribose) polymerase (PARP) pathway. In this study, we investigated the effectiveness of the peroxynitrite decomposition catalyst FP15 on ageing-associated cardiac and vascular dysfunction. Young and ageing rats were treated with vehicle or FP15 intraperitoneally. Using a microtip Millar pressure catheter we performed left ventricular blood pressure analysis to assess systolic and diastolic function. Endothelium-dependent and -independent vasorelaxation of isolated aortic rings were investigated by using acetylcholine and sodium nitroprusside. Ageing animals showed a marked reduction of systolic and diastolic cardiac function and loss of endothelium-dependent relaxant responsiveness of aortic rings. FP15-treatment significantly improved cardiac performance and endothelial function. Immunohistochemical staining confirmed that FP15 effectively reduced nitrosative stress and prevented the activation of PARP in the aortic wall of ageing rats. Our results demonstrate the importance of endogenous peroxynitrite-overproduction in the pathogenesis of ageing-associated cardiovascular dysfunction. Pharmacological decomposition of peroxynitrite by FP15 may represent a novel therapeutic utility to improve cardiac and vascular dysfunction associated with ageing. PMID- 17116321 TI - Inflammaging and anti-inflammaging: a systemic perspective on aging and longevity emerged from studies in humans. AB - A large part of the aging phenotype, including immunosenescence, is explained by an imbalance between inflammatory and anti-inflammatory networks, which results in the low grade chronic pro-inflammatory status we proposed to call inflammaging. Within this perspective, healthy aging and longevity are likely the result not only of a lower propensity to mount inflammatory responses but also of efficient anti-inflammatory networks, which in normal aging fail to fully neutralize the inflammatory processes consequent to the lifelong antigenic burden and exposure to damaging agents. Such a global imbalance can be a major driving force for frailty and common age-related pathologies, and should be addressed and studied within an evolutionary-based systems biology perspective. Evidence in favor of this conceptualization largely derives from studies in humans. We thus propose that inflammaging can be flanked by anti-inflammaging as major determinants not only of immunosenescence but eventually of global aging and longevity. PMID- 17116322 TI - Longevity network: construction and implications. AB - The vast majority of studies on longevity have focused on individual genes/proteins, without adequately addressing the possible role of interactions between them. This study is the first attempt towards constructing a "longevity network" via analysis of human protein-protein interactions (PPIs). For this purpose, we (i) compiled a complete list of established longevity genes from different species, including those that most probably affect the longevity in humans, (ii) defined the human orthologs of the longevity genes, and (iii) determined whether the encoded proteins could be organized as a network. The longevity gene-encoded proteins together with their interacting proteins form a continuous network, which fits the criteria for a scale-free network with an extremely high contribution of hubs to the network connectivity. Most of them have never been annotated before in connection with longevity. Remarkably, almost all of the hubs of the "longevity network" were reported to be involved in at least one age-related disease (ARD), with many being involved in several ARDs. This may be one of the ways by which the proteins with multiple interactions affect the longevity. The hubs offer the potential of being primary targets for longevity-promoting interventions. PMID- 17116323 TI - The role of WRN in DNA repair is affected by post-translational modifications. AB - Werner syndrome (WS) is an autosomal recessive progeroid disease characterized by genomic instability. WRN gene encodes one of the RecQ helicase family proteins, WRN, which has ATPase, helicase, exonuclease and single stranded DNA annealing activities. There is accumulating evidence suggesting that WRN contributes to the maintenance of genomic integrity through its involvement in DNA repair, replication and recombination. The role of WRN in these pathways can be modulated by its post-translational modifications in response to DNA damage. Here, we review the functional consequences of post-translational modifications on WRN as well as specific DNA repair pathways where WRN is involved and discuss how these modifications affect DNA repair pathways. PMID- 17116324 TI - Automatic and controlled processes and the development of addictive behaviors in adolescents: a review and a model. AB - This paper presents a review and a model of the development of addictive behaviors in (human) adolescents, with a focus on alcohol. The model proposes that addictive behaviors develop as the result of an imbalance between two systems: an appetitive, approach-oriented system that becomes sensitized with repeated alcohol use and a regulatory executive system that is not fully developed and that is compromised by exposure to alcohol. Self-regulation critically depends on two factors: ability and motivation to regulate the appetitive response tendency. The motivational aspect is often still weak in heavy drinking adolescents, who typically do not recognize their drinking as problematic. Motivation to regulate use often develops only years later, after the individual has encountered serious alcohol-related problems. Unfortunately, at that point behavioral change becomes harder due to several neurocognitive adaptations that result from heavy drinking. As we document, there is preliminary support for the central elements of the model (appetitive motivation vs. self regulation), but there is a paucity of research directly addressing these mechanisms in human adolescents. Further, we emphasize that adolescent alcohol use primarily takes place in a social context, and that therefore studies should not solely focus on intra-individual factors predicting substance use and misuse but also on interpersonal social factors. Finally, we discuss implications of the model for interventions. PMID- 17116325 TI - Arachidonate 15-lipoxygenase and ubiquitin as fertility markers in boars. AB - Accurate semen analysis is an important issue in the swine industry. We evaluated two candidate fertility marker proteins associated with sperm cytoplasmic droplet (CD), including 15-lipoxygenase (15-LOX) and ubiquitin (UBI) in a controlled single-sire artificial insemination (AI) trial. Ejaculates (n=116) were collected from 18 fertile Large White boars monthly for 8 mo, and analyzed by semi quantitative, densitometry-based Western blotting and flow cytometry with antibodies against 15-LOX and UBI. Data were correlated with farrowing rates (FR) and total numbers of piglets born (TNB) from 1754 AI services by 13 of 18 boars, and compared with a conventional microscopic semen analysis. In semi-quantitative Western blotting, both 15-LOX and UBI were correlated with seasonal changes in the percentage of normal (r=-0.38, P<0.01; r=-0.27, P<0.05, respectively) and CD bearing spermatozoa (r=0.35, P<0.01; r=0.27, P<0.05, respectively). In flow cytometry, UBI and 15-LOX levels showed seasonal changes coinciding with seasonal changes of FR and TNB, representing 13 boars, 88 ejaculates and 1,232 AI services. There were correlations between flow cytometric values of UBI and FR (r=0.31; P<0.05), adjusted FR (r=0.30; P<0.05), TNB (r=-0.38; P<0.01) and adjusted TNB (r=-0.37; P<0.01). Flow cytometric measurements of 15-LOX correlated negatively with TNB (r=-0.33; P<0.05) and adjusted TNB (r=-0.34; P<0.05). These data suggested that boar fertility estimation could be achieved within a group of fertile boars by the use of objectively measurable fertility markers. Flow cytometry appeared more informative and more practical than semi-quantitative Western blotting. This technology could be further optimized for the selection of the most fertile sires in an artificial insemination program. PMID- 17116326 TI - Flexural properties of endodontic posts and human root dentin. AB - OBJECTIVES: To evaluate the flexural modulus and flexural strength of different types of endodontic post in comparison with human root dentin. METHODS: Three different types of fiber-reinforced composite (FRC) posts and three metal posts each comprising 10 specimens (n=10) and 20 dentin bars were loaded to failure in a three-point bending test to determine the flexural modulus (GPa) and the flexural strength (MPa). Three randomly selected fiber posts of each group were evaluated using a scanning electron microscope (SEM) to illustrate the differences in mode of fracture. Data were subjected to a one-way ANOVA to determine significant differences between groups and the Bonferroni t-test multiple comparison was applied to investigate which mean values differed from one another with significance levels of P<0.05. RESULTS: The flexural modulus recorded for the dentin bars was 17.5+/-3.8 GPa. The values for posts ranged from 24.4+/-3.8 GPa for silica fiber posts to 108.6+/-10.7 GPa for stainless steel posts. The flexural strength for dentin was 212.9+/-41.9 MPa, while the posts ranged from 879.1+/-66.2 MPa for silica fiber posts to 1545.3+/-135.9 MPa for cast gold posts. The ANOVA test analysis revealed significant differences between groups (P<0.05) for flexural modulus and flexural strength mean values. SIGNIFICANCE: FRC posts have an elastic modulus that more closely approaches that of dentin while that for metal posts was much higher. The flexural strength of fiber and metal posts was respectively four and seven times higher than root dentin. PMID- 17116327 TI - Bi-directional cell trafficking between mother and fetus in mouse placenta. AB - It is now well established that cells are exchanged between mother and fetus during gestation. It has been proposed that some of these exchanges take place in the placenta, but it has never been demonstrated. Here, we made use of EGFP (Enhanced Green Fluorescent Protein) transgenic mice to precisely visualize the juxtaposition of maternal and fetal tissues at the implantation site, as well as to describe the bi-directional cell trafficking between mother and fetus at different stages of gestation. The influence of genetic differences between mother and fetus on the cell migration was also addressed by studying various types of matings: syngeneic, allogeneic and outbred. The frequency of maternal fetal cell exchanges within the placenta is much higher in syngeneic and allogeneic gestations than in outbred ones. Maternal cells were mainly localized in the labyrinth where they were scattered or sometimes grouped in or near blood spaces. Groups of maternal cells could also be observed in maternal blood sinuses of the spongiotrophoblast. Conversely, fetal cells were organized in rings surrounding maternal blood sinuses in the decidua at 10-12 days of gestation. After day 13, they invaded the decidua. Fetal cells could also be detected in maternal peripheral blood and organs by nested PCR and fluorescence microscopy on cryosections, respectively. This suggests a role in the establishment and maintenance of the maternal tolerance to the fetus. PMID- 17116328 TI - Placental abruption is more frequent in women with the angiotensinogen Thr235 mutation. AB - OBJECTIVE: Obstetrical complications such as preeclampsia, fetal growth restriction, and placental abruption are associated with inadequate placental perfusion. Previous studies have shown that the angiotensinogen (AGT) Thr235 mutation is associated with abnormal remodeling of the uterine spiral arteries and occurs at higher frequencies in preeclampsia. This study was done to evaluate whether the AGT Thr235 mutation increases the risk of placental abruption. MATERIALS AND METHODS: We compared 62 placentas from women who had placental abruption with 240 control patients of similar age and ethnicity. DNA was extracted from paraffin blocks from placentas. AGT Met235Thr mutation status was determined by single fluoresceine labeled probe real-time PCR using a LightCycler system. RESULT: AGT genotypes were divided into three groups: MM (homozygous wild), TT (homozygous mutant), and MT (heterozygous). The constituent ratio of AGT genotype in abrupted placentas (MM 14.5%, MT 43.5%, TT 41.9%) was significantly different from in control group (MM42.5%, MT 39.6%, TT 17.9%) (p<0.001). AGT mutant allele frequency in placental abruption (0.637) was significantly higher than in the control group (0.377) (p<0.001). CONCLUSION: The AGT Thr235 mutation was observed more frequently in placental abruption. AGT Thr235 mutation may be considered a risk factor for placental abruption. PMID- 17116329 TI - Welfare recipients' involvement with child protective services after welfare reform. AB - OBJECTIVE: This study identifies factors associated with child protective services (CPS) involvement among current and former welfare recipients after welfare reform legislation was passed in the US in 1996. METHOD: Data come from the Women's Employment Study, a longitudinal study of randomly selected welfare recipients living in a Michigan city in 1997 (N=541). In order to identify risk factors for CPS involvement among current and former welfare recipients, multinomial logit analyses with 29 independent variables were employed on a trichotomous dependent variable: no CPS involvement, investigation only, and supervision by CPS after investigation. RESULTS: The relationship between work and involvement with CPS differs by work experience prior to welfare reform. As the percentage of months working after welfare reform increased, the risk of being investigated by CPS declined among those with prior work experience but the risk increased among those without prior work experience. However, work variables were not significant predictors of supervision by CPS after an initial investigation. Further, race, cohabitation, childhood welfare receipt, having a learning disability, having a large number of children, being newly divorced, living in a high problem neighborhood, and being convicted of a crime were associated with one's probability of being either investigated or supervised by CPS. CONCLUSIONS: These findings suggest that employment could have increased the stress levels of current or former welfare recipients without prior work experience to the point where they were prone to minor child rearing mistakes that resulted in a CPS investigation, but were not severe enough to warrant opening the case for supervision. Supports should be provided to welfare mothers who are prone to involvement with CPS; expansions in the childcare subsidy and a reduction or delay in work requirements might also help these families. PMID- 17116330 TI - Relationships among childhood maltreatment, PTSD, and health in female veterans in primary care. AB - OBJECTIVE: Women with histories of childhood maltreatment (CM) have higher rates of physical health problems and greater medical utilization compared to women without abuse histories. This study examined whether current post-traumatic stress disorder (PTSD) symptoms mediate the relationship between CM and indicators of physical health and medical utilization in female veterans. METHOD: Respondents were 221 female veterans (56% of the potential sample), who received medical care from the San Diego VA Healthcare System during a 12-month period. Respondents provided self-report information about CM, PTSD symptoms, use of pain medication, and physical symptoms and functioning. Additional information about medical utilization was extracted from respondents' medical charts. Regression based models were conducted to test whether PTSD symptoms mediate the relationships between CM and physical symptoms and between CM and medical utilization. RESULTS: Emotional abuse was associated with poorer role-physical functioning, increased bodily pain and greater odds of using pain medication in the past 6 months. Physical abuse was associated with poorer general health. Contrary to prediction, emotional neglect was associated with better role physical functioning, and CM was not associated with increased healthcare utilization. PTSD was shown to mediate the relationship between emotional and physical abuse and health outcomes. CONCLUSIONS: PTSD, or psychopathology more generally, appears to be an important factor in the negative health impact of CM. Given that several empirically supported interventions are available for PTSD, there may be physical health benefits in early identification and treatment of psychopathology related to CM. PMID- 17116331 TI - Expression of complement regulators and receptors on human NT2-N neurons--effect of hypoxia and reoxygenation. AB - Complement activation can cause tissue damage in cerebral stroke by the release of biologically potent activation products and impaired function of regulatory proteins. We investigated the constitutive and hypoxia-reoxygenation-dependent expression of complement receptor 1 (CD35), membrane cofactor protein (CD46), decay-accelerating factor (CD55), protectin (CD59), and complement C3a and C5a receptors (C3aR and C5aR) on human NT2-N neurons. The effect of hypoxia reoxygenation on C3d-deposition on neurons and endothelial cells was also investigated. NT2-N neurons were examined by cellular enzyme-linked immunosorbent assay and immunofluorescence microscopy. Endothelial cells were examined by flow cytometry. Three hours 1% or 0.1% hypoxia and 21h reoxygenation with 50% AB-serum were used to investigate the effect of hypoxia-reoxygenation on regulators and C3d-deposition. NT2-N neurons expressed significant amounts of CD59 (Clone H19/Clone BRIC229: p=0.000006/p=0.000003), CD46 (p=0.00006), CD55 (p=0.003) and C3aR (p=0.00003). CD35 and C5aR were not significantly expressed. There were no effects of hypoxia-reoxygenation on any of the regulators or receptors after 1% hypoxia and reoxygenation. However, CD55 (p=0.02) was down-regulated after 0.1% hypoxia and subsequent reoxygenation with AB-serum. There were no difference observed in the C3d-deposition during hypoxia-reoxygenation in either neurons or endothelial cells. In conclusion, human NT2-N neurons constitutively express C3aR, CD46, CD55 and, in particular, CD59. The cells may respond to locally produced C3a and, at the same time, be well protected against complement attack. Although severe hypoxia-reoxygenation may down-regulate CD55 expression, it does not seem to influence C3d-deposition. PMID- 17116332 TI - Invasive group A streptococcal infection in injecting drug users and non-drug users in a single UK city. AB - OBJECTIVES: Injecting drug users (IDU) represent an increasing proportion of patients with invasive group A streptococcal (GAS) disease. Our aims were to characterise the clinical presentation and strains causing GAS bacteremia in IDU from a single UK city (Brighton and Hove), and to compare this patient group with non-drug users (non-DU) with GAS bacteremia. METHODS: Consecutive GAS blood culture isolates from twenty-two IDU and twenty-two non-DU presenting to the city hospital were studied. Clinical features, strain emm typing and superantigen toxin genotyping were investigated. RESULTS: GAS invasive disease presented differently in IDU compared to non-DU with a predominance of injection site abscesses and lower mortality in IDU. GAS strains from IDU were predominantly emm82 and emm83 types, which are uncommon in the UK and emm82 strains appeared clonal. The non-DU GAS strains demonstrated a broader range of emm types including most frequently emm1 and emm89. There was no major difference in superantigen gene profile between the isolate groups. CONCLUSION: The distinct presentation of invasive GAS disease in IDU compared with non-DU was associated with distinct emm types, a predominance of abscesses, and low mortality, although the small numbers preclude definitive conclusions. Further study is required to establish if these findings reflect strain differences or epidemiological differences in colonisation patterns and injecting practice. PMID- 17116333 TI - Fibrinogen-beta gene haplotype is associated with mortality in sepsis. AB - OBJECTIVES: Fibrinogen plays a key role in coagulation and inflammation. Transcription of the fibrinogen-beta gene (FGB) is the rate-limiting step in fibrinogen production. Our aim was to determine whether haplotypes of FGB are associated with mortality and organ dysfunction in a cohort of patients with sepsis. METHODS: A prospective cohort of 631 consecutive Caucasian patients with sepsis from a tertiary care medical-surgical ICU were enrolled in a gene association study. Patients were genotyped for three polymorphisms in FGB: -854 G/A, -455 G/A, and +9006 G/A. Haplotypes were inferred using PHASE. The primary outcome was mortality. Secondary outcomes were severity of organ dysfunction as measured by days alive and free (DAF) of organ dysfunction. RESULTS: Haplotype GAA was associated with a significantly lower 28-day mortality (28.9% vs. 36.9% for all other haplotypes, p=0.03). Carriers of two copies of haplotype GAA (vs. one and zero copies) had more DAF of organ dysfunction. In a multivariate analysis, haplotype GAA was an independent predictor for lower mortality (OR=0.66, 95% CI=0.46-0.94, p=0.02). CONCLUSIONS: Haplotype GAA in FGB is associated with lower mortality and lower severity of organ dysfunction. Haplotype GAA encompasses a previously described haplotype -1420A/-854G/-455A/ 249C/-148T/+1690G that is associated with higher fibrinogen levels. PMID- 17116334 TI - Nickel Gluconate-Mercurius Heel-Potentised Swine Organ Preparations: a new therapeutical approach for the primary treatment of pediatric ranula and intraoral mucocele. AB - OBJECTIVE: Many authors consider surgical therapy of pediatric ranula and intraoral mucocele as the election treatment. Recently, an intracystic sclerosing injection with OK-432 has been proposed as a ranula primary treatment. This preliminary study evaluates the effectiveness of the use of Nickel Gluconate Mercurius Heel-Potentised Swine Organ Preparations as the primary treatment of pediatric ranula and intraoral mucocele. METHODS: Eighteen children (9 ranulas, 9 labial mucoceles, 2 lingual mucoceles) were treated with oral administration of Nickel Gluconate-Mercurius Heel-Potentised Swine Organ Preparations D10/D30/D200. RESULTS: Eighty-nine percent ranulas (8 out of 9), 67% labial mucoceles (6 out of 9) completely responded to the therapy. One ranula, that interrupted therapy after only 4 weeks, was subjected to marsupialization in another hospital. A double mucocele case partially responded (one of the two was extinguished), another case incompletely responded, decreasing the size beyond 50%, and just one case, changing volume, resisted the therapy. Lingual mucocele healed at once. Blandin-Nuhn polypoid congenital mucocele responded to the treatment with gradual reabsorption, permitting surgical excision of the atrophic polypoid remnant, without removing glands of origin. No solved case showed recurrence (follow up range: 4-32 months). CONCLUSION: Homotoxicological therapy with Nickel Gluconate Mercurius Heel-Potentised Swine Organ Preparations D10/D30/D200 is an effective primary treatment of pediatric ranula and intraoral mucocele. PMID- 17116335 TI - GABA sets the tempo for activity-dependent adult neurogenesis. AB - GABA, a major inhibitory neurotransmitter in the adult brain, activates synaptic and extrasynaptic GABA(A) receptors, causing hyperpolarization of mature neurons. As in the embryonic nervous system, GABA depolarizes neural progenitors and immature neurons in the adult brain. Several recent studies have suggested that GABA has crucial roles in regulating different steps of adult neurogenesis, including proliferation of neural progenitors, migration and differentiation of neuroblasts, and synaptic integration of newborn neurons. Here, we review recent findings on how GABA regulates adult neurogenesis in the subventricular zone of the lateral ventricles and in the dentate gyrus of the hippocampus. We also discuss an emerging view that GABA serves as a key mediator of neuronal activity in setting the tempo of adult neurogenesis. PMID- 17116336 TI - The acyclic nucleoside phosphonates from inception to clinical use: historical perspective. AB - The collaboration between Antonin Holy [Institute of Organic Chemistry and Biochemistry (IOCB, Prague, Czech Republic)] and Erik De Clercq (Rega Institute for Medical Research, K.U. Leuven, Belgium) started exactly 30 years ago. It led to the discovery of a (rather small) series of acyclic nucleoside analogues (prototype: DHPA), followed by the discovery of a large number of nucleotide analogues [acyclic nucleoside phosphonates (ANPs)] (prototype: HPMPA). From HPMPA originated three compounds, which have been approved by regulatory agencies worldwide for clinical use: (i) HPMPC [cidofovir (Vistide)] for the treatment of cytomegalovirus (CMV) retinitis in AIDS patients, and "off label" for the treatment of polyoma-, papilloma-, adeno-, herpes- and poxvirus infections; (ii) PMEA [adefovir (in its oral prodrug form, adefovir dipivoxil (Hepsera)] for the treatment of chronic HBV (hepatitis B virus) infections, and (iii) PMPA [tenofovir (in its oral prodrug form, tenofovir disoproxil fumarate (Viread)] for the treatment of HIV infections (AIDS). The latter has also been approved, in combination with emtricitabine (Truvada), and in combination with emtricitabine and efavirenz (Atripla((R))) for the treatment of HIV infections. Many other ANPs such as the DAP derivatives HPMPDAP, PMEDAP and PMPDAP, and the DAPy derivatives HPMPO-DAPy, PMEO-DAPy, and PMPO-DAPy, were found to exhibit an antiviral activity spectrum and potency comparable to that of the parent compounds HPMPA (and HPMPC), PMEA and PMPA, respectively. PMID- 17116337 TI - Insulin protects against stress-induced impairments in water maze performance. AB - The presence of insulin receptor in the hippocampus suggests that this organ is a target for insulin. However, unlike the classic peripheral insulin target tissues such as adipocyte, muscle and liver, where the primary function of insulin is to regulate glucose homeostasis, insulin in the central nervous system (CNS) exhibits more diverse actions, most of which have not been clearly understood. A direct role of hippocampal insulin receptor signaling in improving cognitive functions, including learning and memory, and the association of insulin receptor deterioration with brain degenerative dementia (e.g., Alzheimer's disease) have attracted increasing interest. Additionally it has been shown that insulin can be a neuroprotective agent against memory loss induced by ischemia, lesions and some pharmacological agents. In the present study we evaluate the hypothesis that the bilateral intra CA1 insulin injection can protects against stress-induced memory deficit. Chronic restraint stress (2h per day x 7 days) significantly impaired spatial performance in Morris water maze and elevated serum corticosterone level. Intrahippocampal insulin microinjection was done 15-20 min before every stress episode. Insulin in low dose (0.5 MU) had no significant effect on memory deficit induced by stress. But in higher doses (6 and 12 MU) insulin protects animals against the deleterious effect of stress. Insulin alone daily injection had no effect on water maze performance. These results suggest that spatial learning and memory is compromised during chronic stress and insulin may protect against this effect. PMID- 17116339 TI - Measurements of chromogranin B can serve as a complement to chromogranin A. AB - OBJECTIVE: CgA has been shown to be an excellent marker for neuroendocrine tumours. However, there are two major drawbacks with CgA measurements; elevated levels are common in patients with decreased renal function and in patients on treatment with proton pump inhibitors. These problems are not seen with CgB measurements. We have recently presented the development of 13 region-specific radioimmunoassays for measurements of CgB. A region-specific assay was identified, which measured higher concentrations of CgB than the other assays and seemed to be very useful as a marker for neuroendocrine tumours. The aim of the present study was therefore to further explore the diagnostic potential of this assay in the clinical management of patients with neuroendocrine tumours. METHODS: Measurements of CgB with two methods were compared with CgA in plasma samples from patients investigated for neuroendocrine tumours (N=86), patients with decreased renal function (N=35) and patients on treatment with proton pump inhibitors (N=29). RESULTS: The diagnostic sensitivity for the new CgB assay was almost as good as that for CgA. Furthermore, with CgB measurements we could avoid the falsely elevated levels of CgA found in patients with decreased renal function and treatment with proton pump inhibitors. CONCLUSIONS: We conclude that the new CgB assay can serve as a complement to CgA measurements as an important tumour marker for neuroendocrine tumours. PMID- 17116340 TI - Adding synchrotron radiation to infrared microspectroscopy: what's new in biomedical applications? AB - Infrared spectroscopy and microscopy have heralded a period of rapid advances in tissue and cellular characterization during the past decade. However, vibrational spectroscopy is still an analytical tool that is neither familiar nor understood in the medical environment. For many years this field has been mainly driven by physicists and chemists, who are, undoubtedly, at the forefront of tremendous technical developments in technology, detection and data treatment. Although the theory of infrared (IR) spectroscopy is thoroughly worked out, the scientific ground of vibrational spectroscopy is now undergoing a real boost, with the application of this analytical technique in biology and biomedicine. PMID- 17116342 TI - Improving the nutritive value of tubers: elevation of cysteine and glutathione contents in the potato cultivar White Lady by marker-free transformation. AB - The amino acids that limit the nutritive value of potato are the sulfur containing amino acids methionine and cysteine. Manipulation of the targeted amino acid biosynthesis is a way to circumvent this problem. Cysteine is synthesised from O-acetyl-l-serine formed by serine acetyltransferase (SAT). To increase the cysteine content of the commercial potato cultivar White Lady the chimeric SAT-coding cysE gene from Escherichia coli under the control of the constitutive CaMV 35S promoter and fused to the chloroplast targeting rbcS 5' transit peptide sequence was introduced into the White Lady genome. Novelty of the approach was the application of marker-free transformation. Two transgenic lines were obtained that accumulated the cysE mRNA in high amounts. Crude leaf extracts of these plants exhibited up to 80- and 20-fold higher SAT activity in leaves and tubers, respectively, than those prepared from non-transformed plants. Levels of cysteine and glutathione both in leaves and tubers were 1.5-fold higher in average than in control plants. The alterations observed had no effect on tuber yield and sprouting behaviour. Gas chromatography coupled to mass spectrometry showed that all other amino acids than cysteine were unaffected. Here we demonstrate for the first time that the cysteine content of tubers can be enhanced by metabolic engineering. PMID- 17116343 TI - Recent advances in iron oxide nanocrystal technology for medical imaging. AB - Superparamagnetic iron oxide particles (SPIO and USPIO) have a variety of applications in molecular and cellular imaging. Most of the recent research has concerned cellular imaging with imaging of in vivo macrophage activity. According to the iron oxide nanoparticle composition and size which influence their biodistribution, several clinical applications are possible: detection liver metastases, metastatic lymph nodes, inflammatory and/or degenerative diseases. USPIO are investigated as blood pool agents with T1 weighted sequence for angiography, tumour permeability and tumour blood volume or steady-state cerebral blood volume and vessel size index measurements using T2 weighted sequences. Stem cell migration and immune cell trafficking, as well as targeted iron oxide nanoparticles for molecular imaging studies, are at the stage of proof of concept, mainly in animal models. PMID- 17116344 TI - Degradation of the herbicide dichlobenil and its metabolite BAM in soils and subsurface sediments. AB - The worldwide used herbicide dichlobenil (2,6-dichlorobenzonitrile) has resulted in widespread presence of its metabolite 2,6-dichlorobenzamide (BAM) in surface water and groundwater. To evaluate the potential for natural attenuation of this BAM pollution in groundwater, we studied the degradation of BAM and dichlobenil in 16 samples of clayey till, unconsolidated sand and limestone, including sediments from both oxidized and reduced conditions. The degradation of dichlobenil occurred primarily in the upper few meters below surface, although dichlobenil was strongly sorbed to these sediments. However, the degradation of dichlobenil to BAM could not be correlated to either sorption, water chemistry, composition of soils or sediments. Degradation of dichlobenil to BAM was limited (<2% degraded) in the deeper unsaturated zones, and no degradation was observed in aquifer sediments. This illustrates, that dichlobenil transported to aquifers does not contribute to the BAM-contamination in aquifers. A small, but significant degradation of BAM was observed in the upper part of the unsaturated zones in sandy sediments, but no degradation was observed in the clayey till sediment or in the deeper unsaturated zones. The insignificant degradation of BAM in aquifer systems shows that BAM pollution detected in aquifers will appear for a long time; and consequently the potential for natural attenuation of BAM in aquifer systems is limited. PMID- 17116345 TI - HLA-DR association with the genetic susceptibility to develop ashy dermatosis in Mexican Mestizo patients. AB - BACKGROUND: Ashy dermatosis, also known as erythema dyschromicum perstans, is an acquired benign disease, characterized by blue-gray pigment patches with erythematous borders. The cause is still unclear, but probably has an immunologic basis. OBJECTIVE: The aim of this study was to determine gene frequencies of the HLA-DR alleles in Mexican patients with ashy dermatosis and compare them with ethnically matched healthy control subjects to reveal the genetic susceptibility to develop ashy dermatosis. METHODS: We included 23 consecutive patients with clinical and histopathologic confirmed diagnosis of erythema dyschromicum perstans. Patients and control subjects received a questionnaire to determine their ethnic origin and a peripheral blood sample was taken for DNA extraction. Finally, Genetic HLA-DRB1 was performed by polymerase chain reaction sequence specific oligonucleotide reverse dot blot hybridization. RESULTS: Of the 23 patients included in this study, 65% were women and 35% were men. We observed that the disease was located in the trunk in 17 patients (74%) and the upper limbs in 15 patients (65%). The most frequent allele was HLA-DR4 (65%) (pC < 1 x 10(-6), odds ratio = 6.0, 95% confidence interval = 2.8-12.7) whereas in control subjects it was 23%. The most frequent molecular subtype in both patients and healthy control subjects was DRB1( *)0407, being statistically significant after comparing the two groups (pC < 1 x 10(-6), odds ratio = 7.0, 95% confidence interval = 3.1-15.8). LIMITATIONS: Since this is a disease strongly influenced by ethnicity, extrapolation to other ethnic groups is limited. CONCLUSIONS: Many factors influence the ethiopathogenesis of erythema dyschromicum perstans, but it is strongly suggested to have an important genetic susceptibility conferred by genes located within the major histocompatibility complex region. PMID- 17116346 TI - Kainate induces AKT, ERK and cdk5/GSK3beta pathway deregulation, phosphorylates tau protein in mouse hippocampus. AB - Acute treatment with kainate 30 mg/kg (KA) produced behavioral alterations and reactive gliosis. However, it did not produce major death of mouse hippocampal neurons, indicating that concentrations were not cytotoxic. KA caused rapid and temporal Erk phosphorylation (at 6h) and Akt dephosphorylation (1-3 days). Concomitantly, the activation of GSK3beta was increased 1-3 days after KA. After 7 days, a reduction in GSK3beta activation was observed. Caspase-3 activity increased, but to a lesser extent than calpain activation (measured by fluorimetry and calpain-cleaved alpha-spectrin). As calpain is involved in cdk5 activation, and cdk5 is related to GSK3beta, the cdk5/p25 pathway was examined. Results showed that the p25/p35 ratio in KA-injected mice for 3 days was 73.6% higher than control levels. However, no changes in cdk5 expression were detected. Both Western blot and immunohistochemistry against p-Tau(Thr(231)) indicated an increase at this phosphorylated site of tau protein. Indeed an increase in p Tau(Ser(199)) and p-Tau(Ser(396)) was observed by Western blot. Our results demonstrate that tau hyperphosphorylation, induced by KA, is due to an increase in GSK3beta/cdk5 activity in combination with an inactivation of Akt. This indicates that the calpain/cdk5 pathway for tau phosphorylation has a potential role in delayed apoptotic death evoked by excitotoxicity. Moreover, the subsequent activation of caspase and calpain proteases leads to dephosphorylation of tau, thus increasing microtubular destructuration. Taken together, our results provide new insights in the activation of several kinase-pathways implicated in cytoskeletal alterations that are a common feature of neurodegenerative diseases. PMID- 17116347 TI - Modulation of the infant immune responses by the first pertussis vaccine administrations. AB - Many efforts are currently made to prepare combined vaccines against most infectious pathogens, that may be administered early in life to protect infants against infectious diseases as early as possible. However, little is known about the general immune modulation induced by early vaccination. Here, we have analyzed the cytokine secretion profiles of two groups of 6-month-old infants having received as primary immunization either a whole-cell (Pw) or an acellular (Pa) pertussis vaccine in a tetravalent formulation of pertussis-tetanus diphtheria-poliomyelitis vaccines. Both groups of infants secreted IFN-gamma in response to the Bordetella pertussis antigens filamentous haemagglutinin and pertussis toxin, and this response was correlated with antigen-specific IL-12p70 secretion, indicating that both pertussis vaccines induced Th1 cytokines. However, Pa recipients also developed a strong Th2-type cytokine response to the B. pertussis antigens, as noted previously. In addition, they induced Th2-type cytokines to the co-administrated antigen tetanus toxoid, as well as to the food antigen beta-lactoglobulin. Furthermore, the general cytokine profile of the Pa recipients was strongly Th2-skewed at 6 months, as indicated by the cytokines induced by the mitogen phytohaemagglutinin. These data demonstrate that the cytokine profile of 6-month-old infants is influenced by the type of formulation of the pertussis vaccine they received at 2, 3 and 4 months of life. Large prospective studies would be warranted to evaluate the possible long-term consequences of this early modulation of the cytokine responses in infants. PMID- 17116348 TI - How is maternal psychosocial health assessed and promoted in the early postnatal period? Findings from a review of hospital postnatal care in Victoria, Australia. AB - OBJECTIVE: to describe how women's maternal health, particularly at a psychosocial level, is assessed and promoted during the postnatal hospital stay. DESIGN: postal survey of public hospitals providing postnatal care and interviews with care providers. SETTING: all publicly funded maternity units and selected health professionals in Victoria, Australia. PARTICIPANTS: hospital postal survey: sixty six hospital respondents; interviews: 38 maternity unit managers, clinical midwives and medical practitioners. FINDINGS: there was little consistency across the State in relation to routine observations of the mother. Physical checks were much more common than enquiring about how women felt physically. Practice in psychosocial assessment was also diverse, with care plans/maps (clinical pathways) being the main tool to guide assessment. Most participants reported that psychosocial assessment was undertaken during pregnancy. Follow-up after birth also varied. Hospital respondents reported that emotional well-being is assessed postnatally by observation and conversation with women. Participants who were interviewed reported that midwives had mixed skills in assessing and dealing with complex psychosocial issues. Three hospitals administer the Edinburgh Postnatal Depression Scale to women in the days after birth, and three hospitals provide routine sessions of structured debriefing. Survey participants reported that the busy and, at times, chaotic nature of postnatal wards affected the provision of care and the level of psychosocial support offered to women. KEY CONCLUSIONS: although one of the stated aims of early postnatal care is the promotion of maternal well-being, the diversity of practices and the routine nature of many of these practices suggest that care is often not individualised or woman-centred. The reliance of detecting and managing women with particular psychosocial issues during pregnancy results in this aspect of care being given less priority postnatally than may be ideal. IMPLICATIONS FOR PRACTICE: strategies are required to provide health professionals with guidelines and skills to enhance the detection of women who have, or have the potential to develop, health problems after birth. This requires a reorganisation of the way early postnatal care is provided in relation to the use of routine practices; the ability of caregivers to spend time with women in an environment that offers privacy and confidentiality; the structuring of care around individual needs; and opportunities for women to be cared for by caregiver/s with whom they have met before. PMID- 17116349 TI - Psidium guajava 'Paluma' (the guava plant) as a new bio-indicator of ozone in the tropics. AB - Psidium guajava 'Paluma' saplings were exposed to carbon filtered air (CF), ambient non-filtered air (NF), and ambient non-filtered air+40ppb ozone (NF+O(3)) 8h per day during two months. The AOT40 values at the end of the experiment were 48, 910 and 12 895ppbh(-1), respectively for the three treatments. After 5 days of exposure (AOT40=1497ppbh(-1)), interveinal red stippling appeared in plants in the NF+O(3) chamber. In the NF chamber, symptoms were observed only after 40 days of exposure (AOT40=880ppbh(-1)). After 60 days, injured leaves per plant corresponded to 86% in NF+O(3) and 25% in the NF treatment, and the average leaf area injured was 45% in NF+O(3) and 5% in the NF treatment. The extent of leaf area injured (leaf injury index) was explained mainly by the accumulated exposure of ozone (r(2)=0.91; p<0.05). PMID- 17116350 TI - Tissue heavy metal concentrations of stranded California sea lions (Zalophus californianus) in Southern California. AB - Concentrations of nine heavy metals (As, Cd, Cu, Fe, Hg, Pb, Mn, Mo and Zn) were determined in the hepatic and renal tissues of 80 stranded California sea lions (Zalophus californianus). Significant age-dependant increases were observed in liver and kidney concentrations of cadmium and mercury, and renal zinc concentrations. Hepatic iron concentrations were significantly higher in females than males. Animals with suspected domoic acid associated pathological findings had significantly higher concentrations of liver and kidney cadmium; and significantly higher liver mercury concentrations when compared to animals classified with infectious disease or traumatic mortality. Significantly higher hepatic burdens of molybdenum and zinc were found in animals that died from infectious diseases. This is the largest study of tissue heavy metal concentrations in California sea lions to date. These data demonstrate how passive monitoring of stranded animals can provide insight into environmental impacts on marine mammals. PMID- 17116351 TI - Socioeconomic status and health in childhood: a comment on Chen, Martin and Matthews, "Socioeconomic status and health: do gradients differ within childhood and adolescence?" (62:9, 2006, 2161-2170). AB - Understanding whether the gradient in children's health becomes steeper with age is an important first step in uncovering the mechanisms that connect economic and health status, and in recommending sensible interventions to protect children's health. To that end, this paper examines why two sets of authors, Chen et al. [Socioeconomic status and health: Do gradients differ within childhood and adolescence? Social Science & Medicine, 62, 2161-2170.] and Case et al. [Economic status and health in childhood: The origins of the gradient. American Economic Review, 92, 1308-1334.], using data from the same source, reach markedly different conclusions about income-health gradients in childhood. We find that differences can be explained primarily by the inclusion (exclusion) of a small number of young adults who live independently. PMID- 17116352 TI - No association between the TPH A218C polymorphism and personality traits in Japanese healthy subjects. AB - It has been suggested that the central serotonergic activity is implicated in personality traits. Tryptophan hydroxylase (TPH) is the rate-limiting enzyme in serotonin synthesis. In the present study, the association between the TPH A218C polymorphism and personality traits assessed by the Temperament and Character Inventory (TCI) was examined in 345 Japanese healthy subjects. The TPH A218C polymorphism was determined by a PCR-RFLP method. There were no significant differences in the seven dimension scores of TCI among the A/A, A/C, and C/C genotype groups by the one-way ANOVA. There was a significant negative correlation between age and the NS scores. Females showed significantly higher scores of HA, RD, and ST, and significantly lower scores of SD than males. The multiple regression analysis using age, gender, and the TPH genotype as independent variables also showed no significant association between any dimension score and the genotype. The present study thus suggests that the TPH A218C polymorphism does not affect personality traits in Japanese healthy subjects. PMID- 17116353 TI - Pronamel and tooth mousse: an initial assessment of erosion prevention in vitro. AB - OBJECTIVES: The aim of this study was to examine whether a single topical application of proenamel or tooth mousse would prevent enamel erosion METHODS: Enamel samples were treated with either proenamel or tooth mousse applied for 15 min. The control group was placed in distilled water for 15 min. All specimens were then exposed to an erosive challenge of 0.2% citric acid for 1h. Enamel loss was determined using surfometry. RESULTS: The mean amount of enamel removed in the control group was 5.02 microm (S.D. 1.16). The mean enamel loss in the proenamel group was 2.60 microm (S.D. 0.90) and the mean enamel loss in the tooth mousse group was 3.28 microm (S.D. 1.22). The results for the proenamel group were statistically significantly different from the control group at the p<0.01 level and the results for the tooth mousse group were statistically significantly different from the control group at the p<0.05 level. CONCLUSIONS: Tooth mousse and proenamel may offer a degree of protection from erosion of permanent enamel. PMID- 17116354 TI - In vitro FT-IR study of the effects of hydrogen peroxide on superficial tooth enamel. AB - OBJECTIVES: The aim of the present study was to determine the alteration in human enamel after hydrogen peroxide treatment using FT-IR spectroscopy. It is hypothesized that infrared spectroscopy is capable of showing alterations in human enamel after peroxide treatment and the alteration in enamel is proportional to peroxide concentration. METHODS: The effects of 10, 20 and 30% hydrogen peroxide solutions on human enamel were tested. Thirty non-carious human teeth, extracted for periodontal reasons, were used in this study. They were divided into 3 groups of 10, according to the peroxide concentration, sectioned, and the specimens were embedded in resin for infrared spectroscopic analysis. The total treatment time was 120 min. Spectra of the specimens were taken before treatment and 30, 60 and 120 min after it. Another spectrum was taken in a week. RESULTS: Infrared spectroscopic analysis showed two distinct bands (biological PO(4)nu1 and nu2) that were capable of describing the alterations in enamel structure. On comparing the infrared spectra of non-treated and treated specimens, structural changes were detected in the superficial enamel. The alteration in enamel was proportional to treatment time and hydrogen peroxide concentration. Higher concentration and longer treatment time resulted in more severe alterations. The numerical analysis of the spectra revealed that on using concentrated hydrogen peroxide solutions the alterations of the IR spectra were more pronounced. The spectra taken in 1 week after treatment did not show spontaneous reversibility in enamel structure. CONCLUSION: At-home and in-office peroxide-containing bleaching agents are capable of causing alteration in enamel at low and high concentrations as well. According to the results of this study it is recommended to perform tooth whitening using low concentration of hydrogen and/or carbamide peroxide, and shorten treatment time to reduce the possible destruction but reach the required change in color. PMID- 17116355 TI - Molecular modelling studies on the interactions of human DNA topoisomerase IB with pyridoxal-compounds. AB - Candida guilliermondii and human DNA topoisomerases I are inhibited by PL (pyridoxal), PLP (pyridoxal 5'-phosphate) and PLP-AMP (pyridoxal 5'-diphospho-5' adenosine) (PL4%) in slaughtered carcasses. When DNA extracted from piggery bedding materials was analyzed by Mycobacterium-targeted qPCR using the SYBR green I dye for detection of amplification products, 10(5) to 10(7) cell equivalents of mycobacterial DNA were detected in unused bedding materials and 10(8) to 10(10)g( 1) dry weight in used bedding materials. When Mycobacterium-specific hybridization probes were used for detection of amplification products, 10(5) to 10(7) cell equivalents of mycobacterial DNA g(-1) dry weight were detected in unused bedding materials in four out of the five piggeries studied and up to 10(8) cell equivalents in used bedding material. The results were confirmed by the Mycobacterium-specific 16S rRNA sandwich hybridization assay. The present results show, that mycobacteria occur in organic materials commonly used on pig farms, and may proliferate in bedding materials during use. We also show that DNA and RNA-based methods may be utilized for detection of environmental reservoirs of mycobacteria causing porcine and human infection. PMID- 17116376 TI - Estradiol production by preimplantation blastocysts and increased serum progesterone following estradiol treatment in llamas. AB - Estradiol is a potential candidate for the blastocyst signal responsible for maternal recognition of pregnancy in the llama (Lama glama). Two experiments were conducted to determine if the llama blastocyst produces estradiol during the presumed period of maternal recognition of pregnancy and if exogenous estradiol can extend the luteal phase. In Experiment 1, llamas were superovulated with eCG and mated 7 days later (Day 0=day of mating). Blastocysts were collected nonsurgically on Days 7, 9, or 11 or at necropsy on Days 13 and 15 post-mating and cultured for 48h. Conditioned medium was recovered, replaced with fresh medium at 24-h intervals, and assayed for estradiol-17beta. Estradiol production (pg/blastocyst) over the 48-h culture increased (P<0.05) by day of gestation where more estradiol (P<0.05) was produced by Day 11 compared to Day 7 blastocysts, Day 13 compared to Days 7-11 blastocysts, and Day 15 compared to Days 7-13 blastocysts. A dramatic increase was observed between Days 11 and 13 when estradiol production by Day 13 blastocysts increased (P<0.05) more than 50 fold. In Experiment 2, 30 females were induced to ovulate with hCG (Day 0=day of hCG injection). Starting on Day 7 and continuing through Day 15, animals received daily injections i.m. of 0 (n=11), 5 (n=7), or 10mg (n=12) estradiol benzoate (EB) dissolved in isopropylmyristate. Sera were collected immediately prior to each injection and on Days 16, 17, 18, 20, and 22 and analyzed for progesterone. Progesterone concentrations were greater (P<0.05) on Days 14, 15, 16, and 17 in llamas treated with 10mg EB compared to llamas treated with 0mg EB. These results demonstrate that llama blastocysts produce estradiol and exogenous estradiol can enhance and transiently extend luteal progesterone production. Estradiol produced by the preimplantation llama blastocyst may play a role in maternal recognition of pregnancy and early luteal support. PMID- 17116377 TI - ABA and BAB type triblock copolymers of PEG and PLA: a comparative study of drug release properties and "stealth" particle characteristics. AB - We synthesized two types of triblock copolymers containing PEG and PLA as controlled release carriers of hydrophobic drugs: these are the ABA type (PLA-PEG PLA) and the BAB type (PEG-PLA-PEG). These polymers are amphiphilic and can form nanomicelles (40-200nm) in aqueous medium. On the surface of PLA-PEG-PLA (ABA) type nanomicelles, the PEG content was enhanced somewhat over the bulk amount; whereas in the PEG-PLA-PEG (BAB type), surface segregation was much higher. The copolymers tested can entrap 35% of paclitaxel by weight on the average. In general, the diffusion-controlled release of paclitaxel is slower for the BAB polymers; furthermore, the actual release rates are influenced by the PLLA lengths in the BAB copolymers. Surface PEG contents influence the "stealth" characteristics of the nanomicelles. Compared with PLA particles, all nanomicellar particles tested, of both BAB and ABA types, showed a four-fold reduction in monocyte cell uptake, with the BAB type copolymer exhibiting a lesser uptake. PMID- 17116378 TI - Effect of DC/mDC iontophoresis and terpenes on transdermal permeation of methotrexate: in vitro study. AB - The systemic toxicity caused by methotrexate limits its use and transdermal delivery would be a possible alternative. Transdermal permeation of methotrexate loaded into polyacrylamide-based hydrogel patch, across mice skin was studied in vitro after pretreatment with terpenes and ethanol, alone or in combination with iontophoresis (DC/mDC). Polyacrylamide patches gave the maximum flux as compared to the copolymers of acrylamide and acrylic acid. Of the terpenes used, pure menthol showed maximum enhancement (38%), whereas pure limonene elicited a minimum of 9.9% enhancement. Binary combination of menthol and ethanol increased the permeation to 54.9%, which was further enhanced to 93.69% and 117% when used in combination with DC and square wave (mDC) iontophoresis, respectively. ATR FTIR of the stratum corneum treated with terpenes showed a split in the asymmetric C-H stretching vibrations along with decrease in peak heights and areas of asymmetric, symmetric C-H stretching, C=O stretching and amide bands. A split in amide II band was observed with iontophoresis. ATR-FTIR studies suggest conformational changes in the lipid-protein domains thereby increasing permeation. Histopathological studies on treated skin samples, gave an insight about the anatomical changes brought by the application of various enhancers. Binary mixture of menthol and ethanol in combination with square wave gave best results. PMID- 17116379 TI - Novel self-assembling nanogels: stability and lyophilisation studies. AB - The stability of new supramolecular nanoassemblies (nanogels), based on the association of a hydrophobically modified dextran (MD) and a beta-cyclodextrin polymer (pbetaCD), has been studied by two complementary methods: (i) size measurements and (ii) turbidity experiments using a Turbiscan optical analyser. Nanogels of about 120-150nm were obtained whatever the concentration of the two polymer solutions. At low concentrations, the suspensions presented little mean diameter variations upon storage. However, the concentrated ones tended to destabilize and their mean diameter increased upon time. Size measurements and Turbiscan investigations have demonstrated that destabilization in the MD-pbetaCD nanogel suspension was only due to particle aggregation and/or fusion, as no sedimentation or creaming occurred. The destabilization of MD-pbetaCD suspensions led to the formation of a highly viscous phase, as a final state. Moreover, the two methods have shown that aggregation and/or fusion phenomena were more pronounced in the concentrated MD-pbetaCD suspensions than in the diluted ones. The stability of MD-pbetaCD suspensions could be improved by their storage at 4 degrees C. Finally, freeze-drying was found to be a convenient method for the long-time storage of MD-pbetaCD nanoassemblies. PMID- 17116380 TI - Curcumin/turmeric solubilized in sodium hydroxide inhibits HNE protein modification--an in vitro study. AB - Free radical mediated lipid peroxidation has been implicated in multiple diseases. A major oxidation by-product of this deleterious process is 4-hydroxy-2 nonenal (HNE). HNE is cytotoxic, mutagenic and genotoxic and is involved in disease pathogenesis. Curcumin, a non-steroidal anti-inflammatory agent (occurring as the yellow pigment found in the rhizomes of the perennial herb Curcuma longa known as turmeric), has emerged as the newest "nutraceutical" agent that has been shown to be efficacious against colon cancer and other disorders, including correcting cystic fibrosis defects. Since curcumin has been reported to have anti-oxidant properties we hypothesized that it will inhibit HNE modification of a protein substrate. Using an ELISA that employed HNE modification of solid phase antigen following immobilization, we found that the curcumin solubilized in dilute alkali (5mM sodium hydroxide, pH 11) inhibited HNE protein modification by 65%. Turmeric also inhibited HNE-protein modification similarly (65%) but at a much lower alkali level (130muM sodium hydroxide, pH 7.6). Alkali by itself (5mM sodium hydroxide, pH 11) was found to enhance HNE modification by as much as 267%. Curcumin/turmeric has to inhibit this alkali enhanced HNE-modification prior to inhibiting the normal HNE protein modification induced by HNE. Thus, inhibition of HNE-modification could be a mechanism by which curcumin exerts its antioxidant effects. The pH at which the inhibition of HNE modification of substrate was observed was close to the physiological pH, making this formulation of curcumin potentially useful practically. PMID- 17116381 TI - Clinical investigation on spontaneous otoacoustic emission (SOAE) in 447 ears. AB - OBJECTIVES: It is known that spontaneous otoacoustic emission (SOAE) is often observed in normal hearing ears, but concrete clinical application of SOAE test has been rarely reported, compared with transiently evoked otoacoustic emission (TEOAE) and distortion product otoacoustic emission (DPOAE) tests. In addition, there have been a variety of opinions concerning laterality of SOAE, and influence of gender and hearing on SOAE. The reason for this may be that each report has the small number of subjects and lacks in statistical power. Therefore, in the present study, SOAE, TEOAE and DPOAE were measured in 447 ears of subjects at various ages with different hearing level, and statistical analysis was performed to investigate the clinical significance of SOAE. MATERIALS AND METHODS: The subjects were 447 ears in 268 patients (268 ears in females, and 179 ears in males). The age of subjects ranged from 0 to 75 years (mean: 30.8 years), and there were 222 left and 225 right ears. The subjects of schoolchildren or older (414 ears) received pure-tone audiometry, and infants (33 ears) received auditory brain-stem response (ABR). SOAE and TEOAE were measured using ILO88 (Otodynamics, Version 4.20). DPOAE was measured using ILO92 (Otodynamics, Version 1.32). RESULTS: Incidence of SOAE and the number of SOAE per ear were high in the subjects at age of 50 years or younger, in those with hearing level of not more than 30 dB, in the right ear, and in females. Incidence of SOAE in the whole of normal hearing ears was approximately 38%, but the ears with SOAE had almost normal hearing of not more than 30 dB. CONCLUSIONS: SOAE is useful for objective hearing assessment. Moreover, SOAE sometimes appeared in the ears in which TEOAE or DPOAE could not be confirmed, and it might be useful for definite diagnosis of disease state to measure SOAE in addition to TEOAE or DPOAE. PMID- 17116382 TI - Immunoreactivity of calcium binding protein secretagogin in the human hippocampus is restricted to pyramidal neurons. AB - Disturbed calcium homeostasis plays a crucial role in the aetiology of Alzheimer's disease (AD) and the aging process. We evaluated immunoreactivity of secretagogin, a recently cloned calcium binding protein, in hippocampus and adjacent entorhinal cortex of 30 neuropathologically examined post mortem brains (m:f=12:18; mean age, 79.8+/-15.1 years). The study group consisted of 15 cases fulfilling the criteria for high probability of AD according to the NIA-Reagan Institute Criteria and 15 cases with no to medium probability. Sections were incubated with secretagogin-specific antibodies and the number of immunoreactive neurons as well as staining intensities in both neurons and neuropil were assessed. Both cellular and neuropil immunoreactivity were restricted to subiculum and Ammons horn. Cellular immunoreactivity was further restricted to pyramidal neurons and showed a hierarchical distribution: the mean percentage of immunoreactive neurons was highest in sector CA3 (64.41%), followed by CA2 (44.09%), CA4 (34.38%), CA1 (10.9%), and the subiculum (2.92%; P<0.001, except CA2-CA4, P>0.05), while it did not differ significantly between groups with different degrees of AD pathology. The pattern of secretagogin immunoreactivity resembles that of calcium sensor proteins as it is restricted to a subset of neurons and therefore secretagogin could serve highly specialized tasks in neuronal calcium signalling. PMID- 17116384 TI - Effect of melatonin on vascular responses in aortic rings of aging rats. AB - In old animals a marked reduction in endothelium-dependent relaxation occurs. Since there is evidence that the endothelial dysfunction associated with aging may be partly related to the local formation of reactive oxygen species, the purpose of this study was to examine the effect of the natural antioxidant melatonin (10(-5)mol/l) on in vitro contractility of aged aortic rings under conditions of increased oxidative stress (40 m mol/l glucose concentration in medium). Experiments were carried out in 18-20 months old, Wistar male rats, using adult (6-7 months old) animals as controls. A higher plasma lipid peroxidation was found in aged rats as compared to the younger ones. In a first experiment, dose-response curves for acetylcholine-induced relaxation of aortic rings were conducted. Analyzed as a main factor in a factorial ANOVA, age decreased and melatonin augmented the relaxing response to acetylcholine. melatonin's restoring effect on aortic ring relaxation was found in aged aortic rings only and was more pronounced in the presence of a high glucose medium. In a second experiment, the effect of melatonin on the contractility response to phenylephrine of intact or endothelium-denuded aortic rings obtained from aged or control rats was examined in normal or high glucose medium. A main factor analysis in the factorial ANOVA indicated that age and operation augmented, and melatonin decreased, aortic ring contractility response to phenylephrine. Melatonin's restoring effect on aortic contractility was seen in aged aortic rings. The effect of age or a high glucose medium on phenylephrine-induced contractility was more pronounced in the absence of an intact endothelium. Aging did not affect the relaxant response of intact or endothelium-denuded rings to sodium nitroprusside. The results support the improvement by melatonin of vascular response in aging rats, presumably via its antioxidant activity. PMID- 17116385 TI - Simultaneous determination of neomycin sulfate and polymyxin B sulfate by capillary electrophoresis with indirect UV detection. AB - A simple and rapid capillary electrophoresis method, with indirect UV detection, for the simultaneous determination of neomycin sulfate and polymyxin B sulfate in pharmaceutical formulations was developed. Critical parameters such as pH, buffer composition and concentration, voltage and injection time have been studied to evaluate, how they affect responses, such as resolution and migration times. Separation was performed on a fused silica capillary with 50 microm i.d. and 27 cm total length at an applied voltage of 6 kV with a 15 mM phosphate run buffer (pH 5.0) containing 40 mM N-(4-hydroxy-phenyl)acetamide and 50 mM tetradecylammonium bromide (TTAB). The detection wavelength was set at 280 nm. Quantitative analysis was validated by testing the reproducibility of the method, giving a relative standard deviation less than 0.4 and 2.4% for the repeatability of migration time and corrected peak area, respectively. Accuracy was tested by spiking eye-ear formulations with standards and the recoveries of neomycin sulfate and polymyxin B sulfate were found to be between 97.44-103.18% and 96.85 101.68%, respectively. Linearity of neomycin sulfate and polymyxin B sulfate were obtained in the ranges of 17-682 and 24-608 microg/mL, respectively, with r(2) values above 0.999. The established TLC-densitometric method was applied to evaluate the proposed CE method, and comparable results were obtained by using CE with much shorter analysis time and a small quantity of solvents consumed. The developed method is also the first report on the simultaneous determination of neomycin sulfate and polymyxin B sulfate in pharmaceutical preparations by CE. PMID- 17116386 TI - Worries are the mother of many diseases: general practitioners and refugees in the Netherlands on stress, being ill and prejudice. AB - OBJECTIVE: To confront the views of refugee patients and general practitioners in the Netherlands, focusing on medically unexplained physical symptoms (MUPS). METHODS: The study is based on in depth interviews with refugees from Afghanistan (n = 36) and Somalia (n = 30). Additionally, semi-structured interviews were conducted with 24 general practitioners. Text fragments concerning the relationship between mental worries and health or physical ailments were subject of a secondary analysis, the results of which are presented. RESULTS: Medically unexplained physical symptoms were a key issue for both refugees and GPs. The GPs saw MUPS as a significant part of the illness presentation by refugee patients. Refugees felt GPs were often prejudiced, too readily using their difficult background as an explanation for physical symptoms. A 'general narrative' circulating in the refugee communities undermines trust. The GPs applied different strategies in dealing with MUPS presented by their refugee patients. A 'human interest strategy' is distinguished from a 'technical strategy'. The results are discussed in the wider context of the literature on MUPS and patient satisfaction. CONCLUSION: No fundamental difference in paradigms was found between refugees and GPs as to the negative influence worries and bad experiences can have on health. For a fruitful cooperation to develop, based on trust, GPs need to invest in the relationship with individual refugees, and avoid actions based on prejudice. PRACTICE IMPLICATIONS: The importance of (a lack of) trust is underestimated in medical practice. Phenomena undermining trust are often out of sight for practitioners. Critical reflection is needed on the strategies practitioners employ to deal with MUPS. PMID- 17116387 TI - The use of ANAM to assess the side-effect profiles and efficacy of medication. AB - Cognition has become increasingly important as an outcome measure in studies of medication. Traditional neuropsychological assessment is limited in its ability to detect subtle, medication-related changes and it is not suitable for the rapid serial assessment required in most clinical trials. Thus, investigators have turned to computerized neuropsychological assessment for its repeatability, sensitivity to subtle cognitive changes, and ease of administration. The automated neuropsychological assessment metrics (ANAM) is one such computerized battery that has been used to measure the effects of numerous CNS-active drugs. This paper is an exhaustive review of studies that have used ANAM to measure cognitive changes associated with pharmacological treatments. The benefits and limitations of using ANAM in clinical trials are discussed. PMID- 17116388 TI - Inhibitory effects of interferon-gamma on activation of rat pancreatic stellate cells are mediated by STAT1 and involve down-regulation of CTGF expression. AB - Pancreatic stellate cells (PSCs) are the main source of extracellular matrix proteins in pancreatic fibrosis, a pathological feature of chronic pancreatitis and pancreatic cancer. Interferon-gamma (IFN-gamma) is an antifibrotic cytokine, but how precisely it exerts its effects on PSCs is largely unknown. Here, we have focussed on the role of STAT1 as well as target genes of IFN-gamma signalling. Our data indicate that IFN-gamma regulates the expression of two autocrine mediators of PSC activation, connective tissue growth factor and endothelin-1, in a transforming growth factor-beta1-antagonistic manner. STAT1 overexpression under the control of a tetracycline-dependent promoter revealed a close correlation between STAT1 expression and activation, the biological effects of IFN-gamma (growth inhibition, induction of apoptosis), and target gene expression. Our data further support the hypothesis that IFN-gamma interferes with stellate cell activation in the pancreas and suggest activated STAT1 as an inductor of a quiescent PSC phenotype. PMID- 17116389 TI - Afterdischarge threshold reduction in the kindling model of epilepsy. AB - Female Wistar rats were implanted with electrodes in the right amygdala, and initial--pre-kindling--afterdischarge thresholds (ADT's) were determined 1, 2, 4, and 8 weeks after electrode implantation. Pre-kindling ADT's were highest 1 week after implantation, lower at 2 weeks, and lowest at 4 weeks. This drop in non stimulated thresholds had largely disappeared by 8 weeks post-implantation. Half of each group was then kindled, while the other half was "sham" kindled. ADTs were re-measured 1 week after the 10th Stage 5 seizure. When kindling was begun 1 or 2 weeks after implantation (when non-stimulated thresholds were dropping), no effect of kindling on ADT's could be seen. When kindling was begun 4 or 8 weeks after implantation (when non-stimulated thresholds were rising), however, kindling clearly lowered thresholds. PMID- 17116390 TI - CLOCK gene T3111C polymorphism is associated with Japanese schizophrenics: a preliminary study. AB - The CLOCK gene has attracted attention due to its influence on the circadian rhythm, as well as its impacts on the dopaminergic system. We conducted a preliminary study to examine whether the T3111C single nucleotide polymorphism of the CLOCK gene is associated with the development of schizophrenia by examining samples from schizophrenics (n=145) and normal controls (n=128). Both genotype and allele frequencies were significantly different between schizophrenics and controls (p=0.022, p=0.015, respectively). Schizophrenics had a significantly higher frequency of the C allele compared to controls (odds ratio 1.76, 95% CI 1.12-2.75). In particular, disorganized and residual type schizophrenics had significantly higher C allele frequencies than controls (p=0.004 and p=0.037, respectively). Our results suggest that the T3111C polymorphism of the CLOCK gene is associated with schizophrenia. It is important to explore the association between CLOCK and dopamine function, and to examine the impact of CLOCK on phenotypes such as symptoms and drug response in patients with schizophrenia. PMID- 17116391 TI - Endoscopic ultrasound-guided fine needle aspiration in the evaluation of suspected lung cancer. AB - BACKGROUND AND AIM: The role of endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) in the diagnosis and staging of lung cancer is still not fully explored. This prospective study aimed to define the effectiveness of EUS FNA as an adjunct to computer tomography (CT) and bronchoscopy in the evaluation of suspected lung cancer in routine clinical practice. METHODS: Over a period of 20 weeks, the data of 16 consecutive patients suspected of lung cancer on account of respiratory symptoms, and/or the findings of either a mass or mediastinal lymph nodes on helical CT, who were referred for evaluation by EUS, were prospectively collected. Fourteen of these patients underwent sequential bronchoscopy followed by EUS-FNA in the same setting. RESULTS: Bronchoscopy was performed in 15 patients, while EUS was performed in all 16 patients. Bronchoscopy diagnosed 9 cases of non-small-cell lung cancer (NSCLC) but was falsely negative in 3 cases of malignancies, which were all established by EUS FNA of mediastinal lymph nodes (2 cases of NSCLC and 1 case of esophageal squamous cell cancer). EUS-FNA also diagnosed advanced NSCLC in another patient who did not undergo bronchoscopy, such that eventually 13 patients were diagnosed to have malignancies. Distant metastases were diagnosed by EUS-FNA in 4 cases of NSCLC (2 cases of left adrenal gland and 2 cases of pancreatic metastases). Two patients were diagnosed to have sarcoidosis and 1 patient was diagnosed to have pneumoconiosis eventually. CONCLUSIONS: EUS-FNA is useful as an adjunct to CT and bronchoscopy in the evaluation of suspected lung cancer. PMID- 17116392 TI - Sum-frequency generation spectroscopy of DNA monolayers. AB - The anchoring of thiolated single-stranded DNA (HS-ssDNA) monolayers onto platinum substrates was investigated by sum-frequency generation spectroscopy. Different buffer solutions were used for the preparation of the adlayers. Vibrational fingerprints in the 2700-3100 cm(-1) spectral range showed the intercalation of Tris/EDTA (TE) buffer molecules within the HS-ssDNA self assembled monolayer. Buffer contribution to SFG can be quenched either by using SFG inactive molecules like KH(2)PO(4)/K(2)HPO(4)/NaCl (PBS) or by repeated rinsing of the DNA layer with pure water. Comparing the SFG spectra of HS-ssDNA and mercaptohexanol (MCH), which had been self-assembled onto the same substrate, enabled us to infer ordering of the anchor arms and strong disordering of the DNA strands of HS-ssDNA monolayers self-assembled on platinum. PMID- 17116393 TI - Mitochondrial fragmentation in apoptosis. AB - Mitochondrial outer membrane permeabilization (MOMP) is associated with most pro apoptotic stimuli. MOMP results in the release of cytochrome c from the mitochondria into the cytosol, triggering caspase activation and subsequent apoptosis. Several theories explaining the mechanisms of MOMP have been proposed, one of which suggests that MOMP relies on the activation of the molecular machinery involved in fission, resulting in mitochondrial fragmentation. By contrast, several recent studies suggest that mitochondrial fragmentation occurs following MOMP. Moreover, under some conditions. MOMP occurs without mitochondrial fragmentation and, in fact, fragmentation even inhibits MOMP. Here, I discuss the apparently conflicting data and conclude that mitochondrial fragmentation is probably not a prerequisite for MOMP and cytochrome c release. PMID- 17116394 TI - Nursing intensity and patient classification at an adult intensive care unit (ICU). AB - BACKGROUND: ICUs have been using patient information systems for several years now and a huge amount of data is recorded every day. Still nursing research has only limited evidence on how the data recorded in these systems are used. Two of the items systematically recorded in Finnish patient information systems are patient classification and nursing intensity. The aim of this pilot study was to: Use the systematically recorded data of one intensive care unit and discover which variables correlate with or have a statistical relationship to patient classification and nursing intensity. METHODS: The study uses a descriptive statistical approach and the material has been investigated, retrospectively. The material consisted of information system data on 1737 patients at one Finnish intensive care unit in 2003. FINDINGS: Altogether 15,866 patient classifications were made during 1 year. A statistically moderate dependence and a strong correlation (r=0.40) were found between nursing intensity, patient's age, sex and type of intensive care in the case of female emergency non-operative patients and between nursing intensity. That is, the older the female emergency non-operative patient, the higher the assessment of nursing intensity was. Also the correlation between Therapeutic Intervention Scoring System (TISS) scores and nursing intensity was found to be statistically strong (r=0.57). Despite the strong correlation the average TISS score in this study can only account for 32% of the patient's average nursing intensity during the period of treatment. PMID- 17116395 TI - Anomalous centre of mass energy fluctuations during treadmill walking in healthy individuals. AB - Motorised treadmills are used to research and rehabilitate gait despite conflicting evidence that treadmill ambulation is equivalent to ground walking. It has been suggested that no mechanical differences should exist between these environments but there is little evidence to support this. During ground walking, the whole body centre of mass (COM) acts like an inverted pendulum recovering energy, thereby reducing the effort of locomotion. The energy recovery has a relationship with speed whereby maximum recovery occurs at intermediate speeds. In order to determine the relationship between energy recovery and speed during treadmill walking, we investigated estimated COM displacement in nine healthy individuals each walking on a treadmill at seven different speeds. In addition, we measured oxygen cost to determine the effort of walking. Our participants formed two distinct groups, those with normal COM energy recovery (N%R) that was similar to ground walking, and those with low COM energy recovery (L%R) that was different from typical ground walking. The low energy recovery in the L%R group was attributed to in-phase potential and kinetic energy fluctuations. Despite the low energy recovery values both groups produced the expected 'U'-shaped oxygen cost speed curve with no significant difference between groups (p<0.05), however, only N%R produced a significant relationship between energy recovery and oxygen cost (p<0.05). Although a useful tool, walking on a treadmill may not be a true representation of ground walking and therefore not the most effective way to research or rehabilitate gait. PMID- 17116396 TI - Evaluation of [11C]hemicholinium-15 and [18F]hemicholinium-15 as new potential PET tracers for the high-affinity choline uptake system in the heart. AB - [(11)C]Hemicholinium-15 ([(11)C]HC-15) and [(18)F]hemicholinium-15 ([(18)F]HC-15) have been synthesized as new potential PET tracers for the heart high-affinity choline uptake (HACU) system. [(11)C]HC-15 was prepared by N-[(11)C]methylation of the appropriate precursor, 4-methyl-2-phenyl-morpholin-2-ol, using [(11)C]CH(3)OTf in 55-70% radiochemical yield decay corrected to end of bombardment (EOB) and 2-3Ci/mumol specific activity at end of synthesis (EOS). [(18)F]HC-15 was prepared by N-[(18)F]fluoromethylation of the precursor using [(18)F]FCH(2)OTf in 20-30% radiochemical yield decay corrected to EOB and >1.0Ci/mumol specific activity at EOS. The biodistribution of both compounds was determined in rats at 20min post-intravenous injection, and the results show the heart region uptakes 1.32+/-0.75%ID/g in R-ventricle for [(11)C]HC-15 and 1.28+/ 0.81%ID/g in L-ventricle for [(18)F]HC-15, respectively. The dynamic PET imaging studies of [(11)C]HC-15 in rats were acquired 60min post-intravenous injection of the tracer using the IndyPET-II scanner. For the blocking experiments, the rats were intravenously pretreated with 3.0mg/kg of unlabeled HC-15 prior to [(11)C]HC 15 injection. [(11)C]HC-15 rat heart PET studies show rapid heart uptake to give clear heart images. The rat heart PET blocking studies found no significant blocking effect. The dynamic PET studies in normal and ablated dogs were performed using Siemens PET scanner with [(13)N]NH(3), [(11)C]HC-15, and [(18)F]HC-15. PET studies in dogs of both [(11)C]HC-15 and [(18)F]HC-15 also show significant heart uptake and give images of the heart. However, there is no significant change in [(11)C]HC-15 L-ventricle uptake following radiofrequency ablation in the dog. These results suggest that the localization of HC-15 tracers in the heart is mediated by non-specific processes, and the visualization of HC 15 tracers on the heart is related to non-specific binding of HACU. PMID- 17116397 TI - Substrate specificity analysis and inhibitor design of homoisocitrate dehydrogenase. AB - Homoisocitrate dehydrogenase is involved in the alpha-aminoadipate pathway of biosynthesis of l-lysine in fungi, yeast, some prokaryotic bacteria, and archaea. This enzyme catalyzes the oxidative decarboxylation of (2R,3S)-homoisocitrate into 2-oxoadipate using NAD(+) as a coenzyme. Substrate specificity of two homoisocitrate dehydrogenases derived from Deinococcus radiodurans and Saccharomyces cerevisiae was analyzed using a series of synthetic substrate analogs, which indicated a relatively broad substrate specificity of these enzymes. Based on the substrate specificity, 3-hydroxyalkylidene- and 3 carboxyalkylidenemalate derivatives were designed as a specific inhibitor for homoisocitrate dehydrogenase. The synthetic inhibitors showed a moderate competitive inhibitory activity and (R,Z)-3-carboxypropylidenemalate was the most inhibitory among the synthesized inhibitors. Therefore, homoisocitrate dehydrogenase appeared to recognize preferentially an extended conformation of homoisocitrate. PMID- 17116398 TI - The transcervical extended mediastinal lymphadenectomy versus cervical mediastinoscopy in non-small cell lung cancer staging. AB - OBJECTIVE: To compare the diagnostic yield of the transcervical extended mediastinal lymphadenectomy (TEMLA) and the cervical mediastinoscopy (CM) in detecting metastatic mediastinal lymph nodes in NSCLC patients. METHODS: Prospective, randomized, single-blind clinical study. RESULTS: There were 41 NSCLC patients enrolled in the study; 21 were randomized to the TEMLA group and 20 to the cervical mediastinoscopy group. The TEMLA revealed mediastinal metastases in 7 patients, and mediastinoscopy in 3. In the TEMLA group one patient out of the 14 with negative nodes was finally found unfit for surgery, and in the remaining 13 lung resections with mediastinal dissection were performed. In the mediastinoscopy group one patient out of the 17 with negative nodes was finally found unfit for surgery and another one refused surgery, so in 15 of them lung resections with mediastinal dissection were performed. In no patient in the TEMLA group did the pathological examination of the operative specimen reveal metastatic lymph nodes, whereas in the mediastinoscopy group metastatic nodes were found in 5 patients. The number of false negative results was significantly greater in the mediastinoscopy group (5 vs 0, p=0.019), and the difference was the reason for terminating the randomization before reaching the initially planned number of 100 patients. The sensitivity of mediastinoscopy was 37.5% and its negative predictive value was 66.7%, compared to 100% and 100% in the TEMLA group. The comparison of the time of the operation, blood loss, complications, postoperative pain and the use of analgetics has shown significant differences between groups only regarding the operative time and the pain intensity, being greater in the TEMLA group. CONCLUSIONS: 1. The sensitivity and the NPV of the TEMLA in detecting mediastinal metastases in NSCLC are significantly greater than those of cervical mediastinoscopy. 2. The invasiveness of TEMLA and mediastinoscopy does not significantly differ, except for the postoperative pain. PMID- 17116399 TI - Surgical management of bronchiectasis in childhood. AB - OBJECTIVE: Though there is a gradual decrease in the prevalence of bronchiectasis, it is still a cause of mortality and morbidity among children in developing countries such as Turkey. We reviewed the morbidity and mortality rates and the outcome of surgical treatment for childhood bronchiectasis. PATIENTS AND METHODS: Age, sex, etiological factors, symptoms, radiological examinations, surgical procedures, postoperative morbidity and mortality in patients aged 16 years and younger, operated for bronchiectasis between January 1991 and April 2006 in the Thoracic Surgery Clinic of Ataturk Training and Research Hospital for Chest Disease and Chest Surgery were reviewed retrospectively. RESULTS: Between January 1991 and April 2006, 176 cases aged 16 and younger were operated for bronchiectasis. There were 95 females (54%) and 81 males (46%), with a mean age of 12.3 years (range: 3.4-16 years). The most common cause of bronchiectasis was lung infection (n: 87, 49.4%). Main symptoms were coughing (n: 167, 94.9%), sputum (n: 139, 79%), hemoptysis (n: 78, 44.3%), and fever (n: 77, 43.7%). Mean duration of the symptoms in patients who were operated was 3.8 years (range: 0-7.6 years). Cases underwent a total of 201 operations: 19 cases had bilateral surgical resection and 6 cases had completion pneumonectomy. Majority of cases had complete resection (n: 165, 93.75%) while only 11 cases (6.25%) had incomplete resection. Mean duration of hospitalization was 8.9 days (range: 5-39 days). The outcome, based on the responses of patients postoperatively, was 'perfect' in 129 cases (73.3%), 'improved' in 41 cases (23.3%), and 'no changes' in 6 cases (3.4%). The mean follow-up after surgery was 4.3 years (range: 14 months to 7.2 years), mortality was 0% and morbidity was 13% (n: 23). CONCLUSION: With acceptable mortality and morbidity rates and high chance of cure after complete resection, surgical treatment is a successful and reliable method of treatment in childhood bronchiectasis that yields marked improvement in the quality of life. PMID- 17116400 TI - A joined analysis of two European Organization for the Research and Treatment of Cancer (EORTC) studies to evaluate the role of pegylated liposomal doxorubicin (Caelyx) in the treatment of elderly patients with metastatic breast cancer. AB - We have performed a retrospective analysis to evaluate the impact of age, using a 70 year cutoff, on the safety and efficacy of pegylated liposomal doxorubicin (Caelyx) given at 60 mg/m(2) every 6 weeks (treatment A) or 50 mg/m(2) every 4 weeks (treatment B) to 136 metastatic breast cancer patients in two EORTC trials, of whom 65 were 70 years of age or older. No difference in terms of toxicity was observed between younger and older patients treated with the 4-week schedule, while a higher incidence of hematological toxicity, anorexia, asthenia, and stomatitis was observed in older patients when the 6-week schedule was used. Antitumor activity was not affected by age. In the older cohort of patients, no dependence was found between the incidence of grade 3-4 toxicity or antitumor activity and patients' baseline performance status, number and severity of comorbidities, or number of concomitant medications. The higher therapeutic index of Caelyx 50 mg/m(2) every 4 weeks makes it, of the two dose schedules investigated, the preferred regimen in the elderly. PMID- 17116401 TI - Chemically chaperoning the actions of insulin. AB - The role of inflammation as a mediator of insulin resistance in type 2 diabetes and obesity has been a major focus of studies over the past ten years. In mouse models of obesity and type 2 diabetes, the development of insulin resistance correlates with elevated levels of endoplasmic reticulum stress and induction of the unfolded protein response. Activation of N-terminal C-Jun kinase is known to be associated with unfolded protein response activation, and has been shown to participate in the inhibition of insulin action by stimulating serine phosphorylation of the insulin receptor substrate 1, an event that attenuates insulin signaling. 'Chemical chaperones' are small molecules that have been shown to attenuate unfolded protein response activation. The exciting new findings of Ozcan et al. indicate that chemical chaperones improve glucose tolerance and insulin action in a mouse model of type 2 diabetes. These findings offer a potential new target for therapeutic strategies designed to improve insulin action and glucose tolerance in diabetic individuals. PMID- 17116402 TI - Formation of non-random extrachromosomal elements during development, differentiation and oncogenesis. AB - Extrachromosomal elements (EEs) were first discovered as minute chromatin bodies [Cox et al. Minute chromatin bodies in malignant tumors of childhood. Lancet 1965;62:55-8], and subsequently characterized as small circular DNA molecules physically separated from chromosomes. They include episomes, minichromosomes, small polydispersed DNAs or double minutes. This review focuses on eukaryotic EEs generated by genome rearrangements under physiological or pathological conditions. Some of those rearrangements occur randomly, but others are strictly non-random, highly regulated, and involve specific chromosomal locations (V(D)J recombination, telomere maintenance mechanisms, c-myc deregulation). The multiple mechanisms of EEs formation are strongly interconnected and frequently linked to gene amplification. Identification of genes located on EEs will undoubtedly allow a better understanding of genome dynamics and oncogenic pathways. PMID- 17116403 TI - Interpretation of electron density with stereographic roadmap projections. AB - The program RIVEM (Radial Interpretation of Viral Electron density Maps) was developed to project density radially onto a sphere that is then presented as a stereographic diagram. This permits features resulting from an asymmetric reconstruction to be projected and positioned onto an icosahedral virus surface. The features that constitute the viral surface can also be simultaneously represented in terms of atoms, amino acid residues, potential charge distribution, and surface topology. The procedure can also be adapted for the investigation of various molecular interactions. PMID- 17116404 TI - Coarse-grained molecular dynamics simulations of membrane proteins and peptides. AB - Molecular dynamics (MD) simulations provide a valuable approach to the dynamics, structure, and stability of membrane-protein systems. Coarse-grained (CG) models, in which small groups of atoms are treated as single particles, enable extended (>100 ns) timescales to be addressed. In this study, we explore how CG-MD methods that have been developed for detergents and lipids may be extended to membrane proteins. In particular, CG-MD simulations of a number of membrane peptides and proteins are used to characterize their interactions with lipid bilayers. CG-MD is used to simulate the insertion of synthetic model membrane peptides (WALPs and LS3) into a lipid (PC) bilayer. WALP peptides insert in a transmembrane orientation, whilst the LS3 peptide adopts an interfacial location, both in agreement with experimental biophysical data. This approach is extended to a transmembrane fragment of the Vpu protein from HIV-1, and to the coat protein from fd phage. Again, simulated protein/membrane interactions are in good agreement with solid state NMR data for these proteins. CG-MD has also been applied to an M3-M4 fragment from the CFTR protein. Simulations of CFTR M3-M4 in a detergent micelle reveal formation of an alpha-helical hairpin, consistent with a variety of biophysical data. In an I231D mutant, the M3-M4 hairpin is additionally stabilized via an inter-helix Q207/D231 interaction. Finally, CG-MD simulations are extended to a more complex membrane protein, the bacterial sugar transporter LacY. Comparison of a 200 ns CG-MD simulation of LacY in a DPPC bilayer with a 50 ns atomistic simulation of the same protein in a DMPC bilayer shows that the two methods yield comparable predictions of lipid-protein interactions. Taken together, these results demonstrate the utility of CG-MD simulations for studies of membrane/protein interactions. PMID- 17116405 TI - Segmentation of thin structures in electron micrographs using orientation fields. AB - In this paper, we introduce a new approach for segmenting thin structures in electron micrographs. We introduce two new transforms, the Line Filter Transform (LFT) and the Orientation Filter Transform (OFT). The LFT can be viewed as an alternative to anisotropic diffusion algorithms that is particularly useful for thin structures. The OFT utilizes geometrical information about the structure by measuring correlations of local orientations in the image. By combining these methods with a contour extraction and labeling method we construct a segmentation method for thin structures in 2D images. We discuss how the method can be applied slice-by-slice to electron tomograms and illustrate the process by constructing two models of membrane structures from cellular tomograms. The suggested method has the advantage of being relatively insensitive to non-uniform contrast and high-contrast features such as ribosomes. PMID- 17116406 TI - Electric field effects on membranes: gramicidin A as a test ground. AB - Electric fields due to transmembrane potential differences or ionic gradients across the membrane are presumably crucial for many reactions across membranes or close to membranes like signal transduction, control of ion channels or the generation of neural impulses. Molecular dynamics simulations have been used to study the influence of external electric fields on a mixed gramicidin/phospholipid bilayer system. At high field strengths, formation of membrane electropores occurred both close and distal to the gramicidin. Gramicidin was found to stabilize the membrane adjacent to the protein but also at larger distances of up to 2-3 nm. As a result, membrane pore formation was found to be significantly suppressed for the mixed gramicidin/DMPC system. Moderate field strengths only weakly affected the structure and dynamics of the gramicidin. Spontaneous potassium passage events in external electric fields were observed for both the head-to-head helical conformation as well as for the double helical conformation of gramicidin A. The double-helical conformation was found to facilitate ion passage compared to the head-to-head helical dimer. PMID- 17116407 TI - Structural characterisation and expression analysis of toll-like receptor 2 gene from catfish. AB - Toll-like receptors (TLRs) are important components of innate immunity. They were found to recognise specific structures on pathogens termed pathogen-associated molecular patterns (PAMPs) and utilise conserved signaling pathways to activate pro-inflammatory cytokines and type-1 interferons. In spite of much understanding gained from the mammalian systems, many fish TLRs are unknown. Recent studies in Japanese flounder as well as in zebrafish suggested that the ligand binding and activation of inflammatory responses in fish may be different from and more complex than those found in mammals. In channel catfish, the major aquaculture species in the United States, only partial sequences of TLR3 and TLR5 were reported. As a part of efforts to characterise the innate immune components in channel catfish, here we cloned and sequenced both the cDNA and the gene for TLR2, a receptor believed mostly responsible for recognition of lipopeptides on the surface of most Gram-positive bacteria. However, expression analysis after infection with a Gram-negative bacterium, Edwardsiella ictaluri indicated that TLR2 was modestly down-regulated in the head kidney tissue of blue catfish, and with a similar pattern in the head kidney of channel catfish though the down regulation in channel catfish was not statistically significant. In the spleen, an insignificant down-regulation was initially observed early after infection, with an increase of TLR expression later after infection. These results suggest the involvement of TLR2 in the responses after the bacterial infection. As LPS is believed to be the major PAMP for Gram-negative bacteria, additional research is warranted to determine the functions and mechanisms of TLR2 in infections of Gram negative bacteria. PMID- 17116408 TI - Seasonal variation and the immune response: a fish perspective. AB - The environment in which an animal lives affects the physiology and psychology of that animal. The greater the distance from the equator the more profound this influence becomes, as the environment becomes more variable over the years. Temperature, photoperiod, precipitation and other environmental conditions, which are directly or indirectly controlled by the season, can affect an animal. It is becoming apparent that these conditions may impact on the immune system, and this can affect animal health. This review looks at the known mechanisms for transducing environmental cues and how these can affect immune parameters and function. The main focus is fish, especially in relation to aquaculture and the associated disease risks. Work on other animal classes is included for comparison. PMID- 17116409 TI - The influence of interaction type and feeding location on the phylogeographic structure of the yucca moth community associated with Hesperoyucca whipplei. AB - The interactions between herbivorous insects and their host plants have been central in generating diversification in both groups. We used a community of four yucca moth species, monophagous on the host plant Hesperoyucca whipplei (Agavaceae), to examine how the type of interaction and where insects feed within a plant influence phylogeographic structure of herbivorous insects. These four species included two fruit-feeders, one mutualistic and one commensalistic, and two commensalistic stalk-feeders. Surveys based on mtDNA cytochrome oxidase I sequence data demonstrated that the moth species differed in phylogeographic history. Populations of the mutualist pollinator, Tegeticula maculata, exhibited the most subdivision in comparison to the three commensal Prodoxus species (both genera in Lepidoptera, Prodoxidae). Feeding location was also correlated with differences in phylogeographic history through its influence on population sizes and the probability of gene flow. The results suggest that both the outcome of interactions and where insects feed may influence population structure. PMID- 17116410 TI - Molecular evidence for a species complex in the patagonian lizard Liolaemus bibronii and phylogeography of the closely related Liolaemus gracilis (Squamata: Liolaemini). AB - The lizard genus Liolaemus is endemic to temperate South America and includes 190 species. Liolaemus bibronii has a large geographic distribution and inhabits a great diversity of habitats, including the Monte, Steppe, and high Andean grassland environments. Liolaemus gracilis has a similar body size and shape to L. bibronii; the two are parapatrically distributed, and L. gracilis is also widely distributed. Here we use the mtDNA cytb sequence data of these two species to investigate lizard phylogeographic patterns in southern South America. L. bibronii is paraphyletic with respect to L. gracilis, Liolaemus ramirezae, Liolaemus robertmertensi and Liolaemus saxatilis; it is composed of many genetically different allopatric haploclades, some of which are reciprocally monophyletic. We also found evidence for introgression between L. bibronii and L. gracilis in the same area that introgression was hypothesized in the Liolaemus darwinii complex. We discuss the distribution of the major haploclades with inferences of their population histories, the concordance of these clades' distributions and histories with other lizard complexes studied with the same markers and methods, and taxonomic implications of these results. PMID- 17116411 TI - Horizontal gene transfer of a plastid gene in the non-photosynthetic flowering plants Orobanche and Phelipanche (Orobanchaceae). AB - Plastid sequences are among the most widely used in phylogenetic and phylogeographic studies in flowering plants, where they are usually assumed to evolve like non-recombining, uniparentally transmitted, single-copy genes. Among others, this assumption can be violated by intracellular gene transfer (IGT) within cells or by the exchange of genes across mating barriers (horizontal gene transfer, HGT). We report on HGT of a plastid region including rps2, trnL-F, and rbcL in a group of non-photosynthetic flowering plants. Species of the parasitic broomrape genus Phelipanche harbor two copies of rps2, a plastid ribosomal gene, one corresponding to the phylogenetic position of the respective species, the other being horizontally acquired from the related broomrape genus Orobanche. While the vertically transmitted copies probably reside within the plastid genome, the localization of the horizontally acquired copies is not known. With both donor and recipient being parasitic plants, a possible pathway for the exchange of genetic material is via a commonly attacked host. PMID- 17116412 TI - Evolution and diversification of the forest and hypogean ground-beetle genus Trechus in the Canary Islands. AB - The beetle genus Trechus (Carabidae) is represented in the Macaronesian Islands by 43 endemic species. The Canary Islands have 16 endemic species, with two adapted to hypogean life. Phylogenetic relationships among 177 individuals of 38 Canarian, Madeiran, Azorean and continental Trechus species were examined using mitochondrial DNA and nuclear internal transcribed spacer 2 (ITS2) sequence data. Results show two main lineages in the Canaries: one comprising two sister groups with species from the laurel forest of La Gomera and Tenerife, and the other containing the single species from Gran Canaria and a species complex in the four western islands including two troglobites. Calibrations were applied to a linearized tree using a relaxed molecular clock method to estimate the major evolutionary divergence times of the Canarian Trechus species. Although the species assemblage in this archipelago is relatively ancient (7-8 million years), much of the species diversity is recent. Transition to the hypogean environment is more consistent with the "adaptive shift" rather than with the competing "climatic relict" hypothesis. PMID- 17116413 TI - Short-term social recognition memory deficit and atypical social and physiological stressor reactivity in seizure-susceptible El mice. AB - The present studies characterize working memory capabilities in the El mouse model of epilepsy using a species-typical social recognition memory task. As the El mouse exhibits a stress hyper-reactivity phenotype, the impact of hypertonic saline consumption, a memory modulatory treatment, upon social recognition performance was also examined. The hypotheses under test were: (1) that seizure susceptible El mice would perform poorly in the short-term working memory task relative to seizure resistant ddY controls, and (2) that the behavioral and neural responses to stressor exposure would be atypical in El mice. Results revealed a short-term working memory deficit and altered reactivity to social, environmental, and physiological stressors in El mice. In Experiment 1, El mice exhibited poor sociability and decreased olfactory investigation times, both anxiogenic-like traits, compared to ddY controls. In Experiment 2, El mice exhibited poor working memory performance compared to capable performance in ddY controls. Social recognition memory in ddY mice was abolished, however, by salt loading whereas El mice were unaffected by exposure to this physiological stressor. In Experiment 3, all salt-loaded mice exhibited enhanced brain stress neuropeptide (corticotropin releasing factor-CRF) content, and salt-loaded El mice exhibited a 70% reduction in handling-induced seizures. These findings suggest that El mice exhibit high emotionality as well as atypical reactions to stressor exposure, and that these characteristics impact social working memory performance and seizure susceptibility. PMID- 17116414 TI - TRPC6 in glomerular health and disease: what we know and what we believe. AB - Mutations in TRPC6, a member of the transient receptor potential (TRP) superfamily of non-selective cation channels, have been identified as causing a familial form of focal segmental glomerulosclerosis, a disease characterized by proteinuria and progressive renal failure. Here we review the effect of disease associated mutations on TRPC6 function and place TRPC6 within the context of other proteins central to glomerular and podocyte function. Finally, the known roles of TRPC6 in the kidney and other organ systems are used as a framework to discuss possible signaling pathways that TRPC6 may modulate during normal glomerular function and in disease states. PMID- 17116416 TI - Temperature affects insulin-like growth factor I and growth of juvenile southern flounder, Paralichthys lethostigma. AB - Temperature profoundly influences growth of heterothermic vertebrates. However, few studies have investigated the effects of temperature on growth and insulin like growth factor I (IGF-I) in fishes. The aim of this study was to examine effects of temperature on growth and establish whether IGF-I may mediate growth at different temperatures in southern flounder, Paralichthys lethostigma. In two experiments, juvenile flounder were reared at 23 and 28 degrees C and growth was monitored for either 117 or 197 days. Growth was similar across treatments in both experiments until fish reached approximately 100 mm total length. Body size then diverged with fish at 23 degrees C ultimately growing 65-83% larger than those at 28 degrees C. Muscle IGF-I mRNA, plasma IGF-I, and hepatosomatic index (HSI) were significantly higher in flounder at 23 degrees C, whereas hepatic IGF I mRNA abundance did not differ with treatment. Muscle IGF-I mRNA was correlated with HSI, while plasma IGF-I was correlated with body size, hepatic IGF-I mRNA, and HSI. These results demonstrate a strong effect of temperature on flounder growth and show that temperature-induced variation in growth is associated with differences in systemic IGF-I and local (i.e., muscle) IGF-I mRNA levels. The results also support the use of plasma IGF-I and HSI as indicators of flounder growth status. PMID- 17116415 TI - An essential role for FGF receptor signaling in lens development. AB - Since the days of Hans Spemann, the ocular lens has served as one of the most important developmental systems for elucidating fundamental processes of induction and differentiation. More recently, studies in the lens have contributed significantly to our understanding of cell cycle regulation and apoptosis. Over 20 years of accumulated evidence using several different vertebrate species has suggested that fibroblast growth factors (FGFs) and/or fibroblast growth factor receptors (FGFRs) play a key role in lens development. FGFR signaling has been implicated in lens induction, lens cell proliferation and survival, lens fiber differentiation and lens regeneration. Here we will review and discuss historical and recent evidence suggesting that (FGFR) signaling plays a vital and universal role in multiple aspects of lens development. PMID- 17116417 TI - Report of the IWGT working group on strategies and interpretation of regulatory in vivo tests I. Increases in micronucleated bone marrow cells in rodents that do not indicate genotoxic hazards. AB - In vivo genotoxicity tests play a pivotal role in genotoxicity testing batteries. They are used both to determine if potential genotoxicity observed in vitro is realised in vivo and to detect any genotoxic carcinogens that are poorly detected in vitro. It is recognised that individual in vivo genotoxicity tests have limited sensitivity but good specificity. Thus, a positive result from the established in vivo assays is taken as strong evidence for genotoxic carcinogenicity of the compound tested. However, there is a growing body of evidence that compound-related disturbances in the physiology of the rodents used in these assays can result in increases in micronucleated cells in the bone marrow that are not related to the intrinsic genotoxicity of the compound under test. For rodent bone marrow or peripheral blood micronucleus tests, these disturbances include changes in core body temperature (hypothermia and hyperthermia) and increases in erythropoiesis following prior toxicity to erythroblasts or by direct stimulation of cell division in these cells. This paper reviews relevant data from the literature and also previously unpublished data obtained from a questionnaire devised by the IWGT working group. Regulatory implications of these findings are discussed and flow diagrams have been provided to aid in interpretation and decision-making when such changes in physiology are suspected. PMID- 17116418 TI - Theoretical investigation of the molecular, electronic structures and vibrational spectra of a series of first transition metal phthalocyanines. AB - A theoretical investigation of the fully optimized geometries and electronic structures of metallophthalocyanines FePc, CoPc, NiPc, CuPc and ZnPc has been conducted with the density functional theory (DFT) method. A comparison between the different molecules for the geometry, molecular orbital, and atomic charge is made. The simulated order of the sizes of the central hole is FePc>CoPc>NiPcNiPc>CuPc>ZnPc, and the atomic charges of the central metal (M=Fe, Co, Ni, Cu, Zn) ions vary in the same order, FePc>CoPc>NiPcCoPc>FePc>CuPc>ZnPc, and the corresponding peaks predicted at 894, 896, 898, 882 and 871 cm(-1), respectively, also exhibit the same order as above-mentioned. Moreover, the lines of fit through plots of the experimental IR and Ra frequencies versus the calculated ones show very good correlations. PMID- 17116419 TI - Aerobic fitness and field test performance in elite Spanish soccer referees of different ages. AB - The major aim of this study was to examine the physical fitness of elite Spanish soccer referees in relation to their age. A secondary aim was to assess the population criterion validity of the 12 min running test (12 MRT) against aerobic fitness laboratory tests. Participants were 45 soccer referees (age 35.5+/-4.4 years, height 178.3+/-5.0 cm, body mass 75.1+/-6.6 kg, body fat 11.3+/-2.15%, VO2(max), 54.9+/-3.9 ml kg(-1) min(-1)) who were enrolled in the Referees Technical Committee of the Royal Spanish Soccer Federation. They were divided into three age groups: young (Y, 27-32 years, n=15), average (A, 33-38 years, n=17) and old (O, 39-45 years, n=13). No age-related effects were observed for VO2(max), 12 MRT or 200 m sprint performance in either the pooled or grouped data. However, age-related performance decrements were observed for 50 m sprint performance and the ventilatory threshold (VT) running speed. Twelve MRT performance was moderately related to VO2(max), (r=0.46, P=0.002), VT (km h(-1)) (r=0.49, P<0.001), and peak treadmill velocity (PTV) (r=0.60, P<0.001). The results showed that older elite-level referees may be able to limit the expected age-related performance decrements in both aerobic and anaerobic performance usually reported for sedentary people. Additionally, these results show that older referees are able to reach physical fitness levels that have been suggested to be appropriate for coping with match demands. PMID- 17116420 TI - Ambulatory hysteroscopy: evidence-based guide to diagnosis and therapy. AB - Healthcare providers are facing increasing demands for improvement in quality of life for patients. Improvements in service provision for women are being ensured by the introduction of minimally invasive technologies into all spheres of gynaecologic practice. Ambulatory hysteroscopy (direct endoscopic visualization of the endometrial cavity) is an extremely exciting and rapidly advancing field of gynaecologic practice. It advanced dramatically during the 1990s, shifting the focus in healthcare away from inpatient diagnosis and treatment. Hysteroscopy is used extensively in the evaluation of common gynaecological problems that were previously evaluated with blind and inaccurate techniques (e.g. premenopausal menstrual disorders, infertility and postmenopausal bleeding). It allows direct visualization of the uterine cavity and the opportunity for targeted biopsy, safe removal of endometrial polyps, and treatment of submucous fibroids, septa and adhesions. Ambulatory hysteroscopy is safe, with a low incidence of serious complications; it has a small failure rate. There is a general consensus that hysteroscopy is the current gold standard for evaluating intrauterine pathology, including submucous myomas, polyps, hyperplasia and cancer. Hysteroscopy in the ambulatory setting appears to have an accuracy and patient acceptability equivalent to inpatient hysteroscopy under general anaesthetic. The primary goal of this chapter is to provide a high-quality, evidence-based text on ambulatory diagnostic and operative hysteroscopy. The chapter includes in-depth analysis of the indications for outpatient hysteroscopy, its contraindications, the accuracy of diagnostic hysteroscopy, relevant risk management issues and, training and teaching. PMID- 17116421 TI - Testing for ovarian cancer. AB - Ovarian cancer is responsible for more deaths per annum than cervical and endometrial cancer combined. Patients are often diagnosed at a late stage because of the non-specific symptoms of this disease. It can be difficult to differentiate between benign and malignant ovarian pathology, and a malignancy risk index has been developed to guide clinicians. The accuracy of CA125 and ultrasound scans as screening tests is being assessed in randomised controlled trials and proteomic technology shows promise for the early detection of cancers. At present, without accurate screening and early diagnostic techniques, high-risk patients often chose to have prophylactic surgery. PMID- 17116422 TI - Clinical and echocardiographic determinants of ultrasound lung comets. AB - BACKGROUND: Ultrasound lung comets (ULCs) are an echographic sign of extravascular lung water, that originate from water-thickened interlobular septa. AIM: To establish the echocardiographic correlates of ULCs. METHODS: 340 in hospital patients (68 +/- 12years, 115 females) admitted to adult cardiology department underwent upon admission a separate evaluation of chest ULCs and a comprehensive 2D and Doppler echocardiography assessment, including the degree of left ventricular diastolic dysfunction (from 0 = normal to 3 = restrictive pattern). A patient ULC score has been obtained by summing the number of ULCs from each of the scanning spaces in the anterior right and left chest, from second to fifth intercostal space. RESULTS: Multivariate linear regression analysis identified New York Heart Association (NYHA) class (OR = 2.1, CI = 1.4 2.9), ejection fraction (OR 0.954, CI = 0.928-0.981) and degree of diastolic dysfunction (OR = 2.438, CI = 1.418-4.190) as the only parameters independently associated to the number of ULCs. CONCLUSION: ULCs are a simple echographic sign of extravascular lung water, more frequently associated with left ventricular diastolic and/or systolic dysfunction. ULCs can usefully integrate the clinical and pathophysiological information provided by conventional 2D and Doppler echocardiography, in patients with known or suspected heart failure and dyspnoea as a presenting symptom. PMID- 17116423 TI - Laparoscopic transgastric endoscopy after Roux-en-Y gastric bypass. AB - BACKGROUND: Access and endoscopic evaluation of the bypassed stomach is difficult after laparoscopic Roux-en-Y gastric bypass. We propose a minimally invasive technique to access the bypassed stomach after Roux-en-Y gastric bypass for endoscopic diagnosis and treatment. METHODS: First, we established carbon dioxide pneumoperitoneum to a pressure of 12-15 mm Hg. Next, 12-mm umbilical, 5-mm right upper quadrant, 5-mm left lower quadrant, and 15-mm left upper quadrant trocars were placed. A purse-string suture was placed on the anterior wall of the stomach. A gastrotomy was made using ultrasonic shears and the 15-mm trocar was placed into the stomach. The endoscope was then inserted through the 15-mm trocar, and the pneumoperitoneum was decreased to 10 mm Hg. Once the evaluation was complete, the gastrotomy was closed with a running suture or linear stapler. RESULTS: Ten patients at our institution have undergone laparoscopic transgastric endoscopy. Five patients had biliary pathologic findings. Four of these patients underwent successful endoscopic retrograde cholangiopancreatography and papillotomy; the procedure in the fifth patient was unsuccessful because stone impaction at the ampulla. Three patients were evaluated for gastrointestinal bleeding. One was diagnosed with a duodenal gastrointestinal stromal tumor, one with a bleeding duodenal ulcer, requiring surgical exploration; and the third had negative endoscopy findings. Two patients evaluated for chronic abdominal pain had negative endoscopy findings. No complications developed. CONCLUSIONS: Laparoscopic transgastric endoscopy is a safe and minimally invasive approach for the evaluation of the gastric remnant, duodenum, and biliary tree in patients who have undergone Roux-en-Y gastric bypass. PMID- 17116424 TI - Comparison of effects of gastric bypass and biliopancreatic diversion with duodenal switch on weight loss and body composition 1-2 years after surgery. AB - BACKGROUND: Gastric bypass (GB) is the most common surgical procedure for weight loss in the United States. Biliopancreatic diversion with duodenal switch (BPD/DS) is less routinely performed, perhaps because of its technical difficulty and metabolic concerns. The objective of this study was to determine whether these procedures had differential effects on weight loss and body composition. METHODS: Body composition was measured by bioimpedance (Tanita 310) at the initial consultation, and follow-up measurements were completed 1-2 years after surgery. RESULTS: Of the 72 patients in the study, 50, aged 46.2 +/- 8.5 years, had undergone GB and were measured 15.5 +/- 5.2 months after surgery and 22, aged 40.6 +/- 7.9 years, had undergone BPD/DS and were measured 19.5 +/- 7.5 months after surgery. Patient age and time after surgery were significantly different between the 2 groups. The body mass index (BMI) for the BPD/DS group was 53.6 +/- 11.9 kg/m(2), significantly greater than the BMI of the GB group (48.0 +/- 6.3 kg/m(2); P = .009). However, the percentage of body fat did not differ between the 2 groups (P = .515). Postoperatively, the BMIs for the GB group (31.5 +/- 5.0 kg/m(2)) and BPD/DS group (30.3 +/- 6.1 kg/m(2)) were not significantly different (P = .384). The percentage of body fat for the GB and BPD/DS groups had changed from 49.2% +/- 8.3% to 32.1% +/- 10.6% and 47.9% +/- 5.9% to 23.8% +/- 10.4%, respectively (P = .002). The BMI had decreased by 16.5 +/- 4.8 kg/m(2)after GB and 23.3 +/- 6.8 kg/m(2) after BPD/DS (P <.001). The decrease in fat was 17.1% +/ 8.2% after GB and 24.2% +/- 7.2% after BPD/DS (P <.001). CONCLUSION: The BPD/DS procedure is more effective in reducing the BMI and promoting fat loss than is GB. The assessment of the impact of these two operations on an individualized basis offers additional information to assist in the evaluation of these procedures. PMID- 17116426 TI - Anatomic findings and outcomes after antireflux procedures in morbidly obese patients undergoing laparoscopic conversion to Roux-en-Y gastric bypass. AB - BACKGROUND: To report the anatomic findings and outcomes in patients undergoing laparoscopic takedown of Nissen fundoplication with conversion to Roux-en-Y gastric bypass. METHODS: We reviewed the records of patients who underwent laparoscopic Nissen takedown with conversion to Roux-en-Y gastric bypass from January 2001 to April 2006 at the University of Minnesota Medical Center. RESULTS: Eleven patients were identified. Nine patients had gastroesophageal reflux disease preoperatively, of whom six underwent 24-hour pH monitoring. In 2 patients, the pH study findings were negative. Eight prior procedures had been performed laparoscopically. Eight patients were women. The mean age was 44 years. The average body mass index preoperatively was 44 kg/m(2) (range 35-61). The mean follow-up was 13.8 months (range 4-39). The body mass index at follow-up was 30.2 kg/m(2). The operative time was 349 minutes (range 222-624). The hospital length of stay was 3.4 days (range 2-6). No conversions to open surgery were required. No major short-term complications developed. Minor complications included wound or drain site infection in 3 patients, with abscess in 2, pressure sore of the lateral aspect of the foot in 1, pneumonia in 2, and marginal ulcer in 2 patients. No strictures were observed. One internal hernia occurred. Of the 9 patients with gastroesophageal reflux disease preoperatively, all had 100% improvement in symptoms, with complete resolution in 7 (78%). Wrap disruption was present in 5 (45%) of 11 patients. Herniation of an intact wrap had occurred in 1 patient. One patient had both herniation and wrap disruption. CONCLUSION: Laparoscopic conversion of Nissen fundoplication to Roux-en-Y gastric bypass is a feasible salvage operation for recurrent gastroesophageal reflux disease in the morbidly obese. The incidence of wrap disruption appears to be relatively high and the incidence of intact wrap herniation low in obese patients after failed Nissen fundoplication, suggesting that the mechanism of failure after primary antireflux surgery in obese patients may be different than that in normal-weight patients. PMID- 17116427 TI - Vertical banded gastroplasty versus adjustable gastric banding: prospective long term follow-up study. AB - BACKGROUND: Vertical banded gastroplasty (VBG) has been in clinical use since 1979 and adjustable gastric banding (AGB) since 1985. Because promising results were achieved with the adjustable gastric bands available in the market, some surgeons came to the conclusion that VBG might be entirely abandoned and replaced by the adjustable gastric band. The aim of this study was to compare the long term outcome of the 2 restrictive procedures. METHODS: Within a 7-year period (1994-2001), 1117 gastric restrictive procedures were performed in the course of a prospective nonrandomized comparative trial. We report the outcomes of 563 VBG and 554 AGB procedures performed by 2 surgeons. The mean body mass index was 46.9 +/- 09.9 kg/m(2) for VBG and 46.7 +/- 07.8 kg/m(2) for AGB. Patient selection was performed by acceptance by 1 of the 2 surgeons. VBG was performed by laparotomy and AGB using laparoscopy. The Bariatric Analysis and Reporting Outcome System (BAROS) was used to evaluate the postoperative health status and quality of life. RESULTS: The mean duration of follow-up was 92 months (range 60-134), with a minimum of 5 years. The overall follow-up rate was 92%. In the short-term 3-year follow-up, no statistically significant difference was registered between AGB and VBG in terms of weight loss, reduction of co-morbidities, or improvement in quality of life. The 30-day mortality rate was .4% (2 patients) for VBG and .2% (1 patient) for AGB. The overall reintervention rate in the long term was 49.7% for VBG and 8.6% for AGB (P <.0001, odds ratio .0937, 95% confidence interval .065-.133), the reoperation rate was 39.9% for VBG and 7.5% for AGB (P <.0001). The excess weight loss was significantly greater in the VBG group after 12 months (58% for VBG versus 42% for AGB, P <.05). At long-term follow-up (mean 92 months), no significant difference in weight loss was registered between the 2 study groups (59% for VBG and 62% for AGB, P = .923). The BAROS score in the short term (3 years) was good to excellent in 94% and 90% of the VBG and AGB groups, respectively. In the long-term follow-up period, the BAROS score was significantly in favor of the AGB group (83.9% versus 57.8%, P <.0001, odds ratio 3.797, 95% confidence interval 2.072-7.125). The overall resolution rate of co morbidities was 80% in both groups. CONCLUSION: This long-term follow-up study shows that VBG and AGB are effective restrictive procedures to achieve weight loss, and loss of co-morbidities. A statistically significant lower re intervention and re-operation rate and an improved health status and quality of life were registered for AGB. PMID- 17116428 TI - The enigmatic thymine DNA glycosylase. AB - When it was first isolated from extracts of HeLa cells in Josef Jiricny's laboratory, the thymine DNA glycosylase (TDG) attracted attention because of its ability to remove thymine, i.e. a normal DNA base, from G.T mispairs. This implicated a function of DNA base excision repair in the restoration of G.C base pairs following the deamination of a 5-methylcytosine. TDG turned out to be the founding member of a newly emerging family of mismatch-directed uracil-DNA glycosylases, the MUG proteins, that act on a comparably broad spectrum of base lesion including G.U as the common, most efficiently processed substrate. However, because of its apparent catalytic inefficiency, some have considered TDG a poor DNA repair enzyme without an important biological function. Others have reported 5-meC DNA glycosylase activity to be associated with TDG, thrusting the enzyme into limelight as a possible DNA demethylase. Yet others have found the glycosylase to interact with transcription factors, implicating a function in gene regulation, which appears to be critically important in developmental processes. This article reviews all these developments in view of possible biological functions of this multifaceted DNA glycosylase. PMID- 17116429 TI - Uracil in DNA--general mutagen, but normal intermediate in acquired immunity. AB - Deamination of cytosine in DNA results in mutagenic U:G mispairs, whereas incorporation of dUMP leads to U:A pairs that may be genotoxic directly or indirectly. In both cases, uracil is mainly removed by a uracil-DNA glycosylase (UDG) that initiates the base excision repair pathway. The major UDGs are mitochondrial UNG1 and nuclear UNG2 encoded by the UNG-gene, and nuclear SMUG1. TDG and MBD4 remove uracil from special sequence contexts, but their roles remain poorly understood. UNG2 is cell cycle regulated and has a major role in post replicative removal of incorporated uracils. UNG2 and SMUG1 are both important for prevention of mutations caused by cytosine deamination, and their functions are non-redundant. In addition, SMUG1 has a major role in removal of hydroxymethyl uracil from oxidized thymines. Furthermore, UNG-proteins and SMUG1 may have important functions in removal of oxidized cytosines, e.g. isodialuric acid, alloxan and 5-hydroxyuracil after exposure to ionizing radiation. UNG2 is also essential in the acquired immune response, including somatic hypermutation (SHM) required for antibody affinity maturation and class switch recombination (CSR) mediating new effector functions, e.g. from IgM to IgG. Upon antigen exposure B-lymphocytes express activation induced cytosine deaminase that generates U:G mispairs at the Ig locus. These result in GC to AT transition mutations upon DNA replication and apparently other mutations as well. Some of these may result from the generation of abasic sites and translesion bypass synthesis across such sites. SMUG1 can not complement UNG2 deficiency, probably because it works very inefficiently on single-stranded DNA and is down-regulated in B cells. In humans, UNG-deficiency results in the hyper IgM syndrome characterized by recurrent infections, lymphoid hyperplasia, extremely low IgG, IgA and IgE and elevated IgM. Ung(-/-) mice have a similar phenotype, but in addition display dysregulated cytokine production and develop B cell lymphomas late in life. PMID- 17116430 TI - Oxidative DNA damage repair in mammalian cells: a new perspective. AB - Oxidatively induced DNA lesions have been implicated in the etiology of many diseases (including cancer) and in aging. Repair of oxidatively damaged bases in all organisms occurs primarily via the DNA base excision repair (BER) pathway, initiated with their excision by DNA glycosylases. Only two mammalian DNA glycosylases, OGG1 and NTH1 of E. coli Nth family, were previously characterized, which excise majority of the oxidatively damaged base lesions. We recently discovered and characterized two human orthologs of E. coli Nei, the prototype of the second family of oxidized base-specific glycosylases and named them NEIL (Nei like)-1 and 2. NEILs are distinct from NTH1 and OGG1 in structural features and reaction mechanism but act on many of the same substrates. Nth-type DNA glycosylases after base excision, cleave the DNA strand at the resulting AP-site to produce a 3'-alphabeta unsaturated aldehyde whereas Nei-type enzymes produce 3'-phosphate terminus. E. coli APEs efficiently remove both types of termini in addition to cleaving AP sites to generate 3'-OH, the primer terminus for subsequent DNA repair synthesis. In contrast, the mammalian APE, APE1, which has an essential role in NTH1/OGG1-initiated BER, has negligible 3'-phosphatase activity and is dispensable for NEIL-initiated BER. Polynucleotide kinase (PNK), present in mammalian cells but not in E. coli, removes the 3' phosphate, and is involved in NEIL-initiated BER. NEILs show a unique preference for excising lesions from a DNA bubble, while most DNA glycosylases, including OGG1 and NTH1, are active only with duplex DNA. The dichotomy in the preference of NEILs and NTH1/OGG1 for bubble versus duplex DNA substrates suggests that NEILs function preferentially in repair of base lesions during replication and/or transcription and hence play a unique role in maintaining the functional integrity of mammalian genomes. PMID- 17116431 TI - A role for Xrcc2 in the early stages of mouse development. AB - Xrcc2 is one of a family of five Rad51-like genes with important roles in the repair of DNA damage by homologous recombination (HR) in mammals. We have shown previously that loss of Xrcc2 in mice results in severe but variable developmental defects and embryonic lethality, potentially linked to excessive apoptosis. To look at the causes of lethality, and possibly to allow Xrcc2-/- mice to survive to birth, we have produced double knockout mice deficient in either the p53 oncoprotein or Ataxia telangiectasia mutated (Atm). Overall we show that the excessive apoptosis observed in Xrcc2-/- embryos is p53-dependent, and that loss of p53 can restore growth capacity to Xrcc2-/- fibroblasts in culture, but that it cannot rescue the embryonic lethality. Additionally, although the Xrcc2-/- Trp53-/- embryos show a near-normal morphology they remain relatively small in size. Loss of Atm in an Xrcc2-/- embryo has little effect, suggesting that response to loss of HR capacity is not mediated through the Atm kinase in the early stages of mouse development. Further, as seen by reduced expression of the early developmental marker, Delta-like1, the normal developmental programme is perturbed in Xrcc2-/- embryonic tissues, particularly during neurogenesis and somitogenesis. Taken together our data suggest that the accumulation of spontaneous damage in HR-deficient embryos has severe consequences for the development and survival of mammals due to the unregulated loss of cells important to the developmental programme. PMID- 17116432 TI - Protein folding liquid chromatography and its recent developments. AB - The ultimate goal of proteomics is to identify biologically active proteins and to produce them using biotechnology tools such as bacterial hosts. However, proteins produced by Escherichia coli must be refolded to their native state. Protein folding liquid chromatography (PFLC) is a new method developed in recent years, and it is widely used in molecular biology and biotechnology. In this paper, the new method, PFLC is introduced and its recent development is reviewed. In addition the paper includes definitions, advantages, principles, applications for both laboratory and large scales, apparatus, and effecting factors of PFLC. In addition, the role of this method in the future is examined. PMID- 17116433 TI - Determination of perospirone by liquid chromatography/electrospray mass spectrometry: application to a pharmacokinetic study in healthy Chinese volunteers. AB - Perospirone is a novel atypical antipsychotic with a unique combination of 5 HT(1A) receptor agonism as well as 5-HT(2A) and D(2) receptor antagonism. A simple rapid and selective LC-MS method utilizing a single quadrupole mass spectrometer was developed and validated for the determination of perospirone hydrochloride in human plasma. N-hexane was used to extract perospirone hydrochloride and amlodipine benzenesulfonate (internal standard (IS)) from an alkaline plasma sample. LC separation was performed on a XTerra MS C(18) column (100mmx2.1mm, i.d. 3.5microm) using methanol -10mM ammonium acetate (84:16, v/v) as a mobile phase. The quantification of target compounds was obtained by using a selected ion monitoring (SIM) at m/z 427.5 [M+H](+) for perospirone hydrochloride, and at m/z 431.4 [M+Na](+) for IS (amlodipine benzenesulfonate). Perospirone and IS eluted as sharp, symmetrical peaks with retention times of 3.11+/-0.01min and 4.15+/-0.2min, respectively. Calibration curves of perospirone hydrochloride in human plasma at concentrations ranging from 0.10 to 21.1ng/mL exhibited excellent linearity (r(2)=0.9997). The mean absolute recovery of the drug from plasma was more than 85%. Intra- and inter-day relative standard deviations were less than 6.43% and 11.9% for perospirone hydrochloride at the range from 0.32 to 10.6ng/mL. Stability characteristics of the drug-containing plasma were thoroughly evaluated to establish appropriate conditions to process, store and prepare for chromatographic analysis without inducing significant chemical degradation. The following pharmacokinetic parameters were elucidated after administering a single dose of 8mg perospirone hydrochloride. The area under the plasma concentration versus time curve from time 0 to 24h (AUC(0-24)) was 15.48+/-4.23microg/Lh; peak plasma concentration (C(max)) was 2.79+/ 0.78microg/L; time to C(max) (T(max)) was 1.79+/-0.45h; and elimination half-life (t(1/2)) 6.78+/-1.38h. The described assay method showed acceptable precision, accuracy, linearity, stability, and specificity and can be used for pharmacokinetic studies, therapeutic drug monitoring, and drug abuse screening. PMID- 17116434 TI - Traces of phosgene in chloroform: consequences for extraction of anthracyclines. AB - Chloroform is commonly used to extract anthracyclines from various biological matrices. However, their determination can be seriously compromised by phosgene traces present as a result of failing stabilization of chloroform. Out of the three varieties in which chloroform exists (not stabilized, stabilized with an alcohol and stabilized with a hydrocarbon) only the ethanol stabilized type minimizes chances on creating artifacts. Chromatographic separation after extraction of four anthracyclines (doxorubicin, epirubicin, daunorubicin and idarubicin) and two metabolites (13-S-dihydrodoxorubicin and 13-S dihydroepirubicin) with chloroform under various conditions indicate that the appropriate choice of stabilizer in this extraction solvent is highly relevant. PMID- 17116436 TI - Antibiotics--past, present, and future. AB - The drug therapy for diseases other than those caused by microbial agents involves treating the host. In infectious diseases therapy, the goal is to rid the host of the pathogen. Hence, drug therapy is aimed at the pathogen. Because a second living agent is involved in the triangle, drug therapy is affected by the pathogen's nature, its tissue specificity, and, most importantly, the changes it undergoes to survive. The history of antimicrobial therapy has clearly demonstrated that the drugs that are used to treat infections are also responsible for making them more difficult to treat in future. The only way to keep antimicrobial agents useful is to use them appropriately and judiciously. PMID- 17116437 TI - In vitro susceptibility testing versus in vivo effectiveness. AB - The clinical relevance of susceptibility testing has always been questioned because of the difficulty of correlating in vitro susceptibility testing with in vivo clinical effectiveness. Clearly there have always been host/pathogen factors that influence the clinical outcome that cannot be predicted by the results of susceptibility testing. However, improved understanding of pharmacodynamic and pharmacokinetic parameters has greatly improved the use of antimicrobial agents. Most importantly, the integration of these pharmacodynamic and pharmacokinetic indices has greatly improved the correlation between in vitro susceptibility testing and in vivo clinical effectiveness and allows more realistic breakpoints. Finally, the clinical microbiology laboratory has advanced with improved methods as well as the adaptation of breakpoints that are more realistic. PMID- 17116438 TI - New uses for older antibiotics: nitrofurantoin, amikacin, colistin, polymyxin B, doxycycline, and minocycline revisited. AB - Nitrofurantoin, amikacin, colistin, polymyxin B, doxycycline, and minocycline are antibiotics with proven effectiveness against selected pathogens. These antibiotics have not developed resistance over time. As "low-resistance potential antibiotics" that are effective against an increasing number of infections due to resistant gram-positive or gram-negative pathogens, these antimicrobials remain an important part of the antibiotic armamentarium. They will be used increasingly in the future, as highly resistant organisms continue to be important clinically and therapeutic options remain limited. PMID- 17116439 TI - Macrolide and ketolide resistance with Streptococcus pneumoniae. AB - Antimicrobial agents in the macrolide family have long been considered drugs of potential utility in the management of infections caused by Streptococcus pneumoniae. However, with the emergence of macrolide resistance, the clinical value of macrolides in pneumococcal infections is threatened. In part, as a consequence of the development of macrolide resistance, recently the first agent in the ketolide antimicrobial class, telithromycin, was developed and introduced into clinical practice. The ketolides are macrolide antimicrobials whose chemistry has been modified so as avoid the effects of the most common mechanisms of macrolide resistance with S pneumoniae. This discussion reviews the current state of resistance to macrolides and ketolides with S pneumoniae in North America. PMID- 17116440 TI - Antibiotic therapy for Helicobacter pylori. AB - Helicobacter pylori is one of the most common bacterial infections in the world. H pylori infection of the gastric mucosa is the most common cause of peptic ulcers and is believed to be responsible for 50% to 60% of all gastric carcinomas. This infection is difficult to treat because the bacterium is located within the gastric lumen in the mucus and not within the gastric tissue. Antimicrobial therapy for H pylori includes two or three antibiotics plus either a proton pump inhibitor or a histamine receptor antagonist. H pylori readily develops resistance to antibiotics; therefore, if the initial treatment is unsuccessful, repeat treatment should include different antibiotics. PMID- 17116441 TI - Antimicrobial therapy of Clostridium difficile-associated diarrhea. AB - Clostridium difficile-associated diarrhea (CDAD) is the most common etiologically defined cause of hospital-acquired diarrhea. Caused by the toxins of certain strains of C difficile, CDAD represents a growing concern, with epidemic outbreaks in some hospitals where very aggressive and difficult-to-treat strains have recently been found. Incidence of CDAD varies ordinarily between 1 to 10 in every 1,000 admissions. Evidence shows that CDAD increases morbidity, length of stay, and costs. This article described the clinical manifestations of CDAD, related risk factors, considerations for confirming CDAD, antimicrobial and non antimicrobial treatment of CDAD, and issues related to relapses. The article concludes with a discussion of recent epidemic outbreaks involving CDAD. PMID- 17116442 TI - Antimicrobial therapy of multidrug-resistant Streptococcus pneumoniae, vancomycin resistant enterococci, and methicillin-resistant Staphylococcus aureus. AB - Antibiotic resistance among pneumococci, enterococci, and staphylococci has become increasingly important in recent decades. Clinicians should be familiar with the nuances of antibiotic susceptibility testing and interpretation in selecting antibiotics for these infections. The clinical significance of penicillin-resistant Streptococcus pneumoniae, macrolide-resistant S pneumoniae, and multidrug-resistant S pneumoniae is discussed. The clinical spectrum and therapeutic approach to Enterococcus faecalis (i.e., vancomycin-sensitive enterococci) and E faecium (i.e., vancomycin-resistant enterococci) are discussed. Differences in therapeutic approach between methicillin-sensitive Staphylococcus aureus and methicillin-resistant S aureus (MRSA) infections are reviewed. Differences between in vitro susceptibility testing and in vivo effectiveness of antibiotics for hospital-acquired MRSA (HA-MRSA) are described. Finally, the clinical features of infection and therapy of HA-MRSA and community acquired MRSA (CA-MRSA) infections are compared. PMID- 17116443 TI - Monotherapy versus combination therapy. AB - The science of antibiotic therapy for infectious diseases continues to evolve. In many instances where empiric coverage is necessary, treatment with more than one agent is considered prudent. If an etiology is identified, antibiotics are modified based on culture and susceptibility data. Even when the organism is known, more than one antibiotic may be needed. Decisions about antibiotics should be made after assessments of pertinent clinical information, laboratory and microbiology information, ease of administration, patient compliance, potential adverse effects, cost, and available evidence supporting various treatment options. Clinicians also need to consider synergy and local resistance patterns in selecting therapeutic options. In this article, the authors outline monotherapy and combination therapy options for several common infectious diseases. PMID- 17116444 TI - Oral antibiotic therapy of serious systemic infections. AB - Traditionally, antibiotics have been administered intravenously (IV) for serious systemic infections. As more potent oral antibiotics were introduced, and their pharmacokinetic aspects studied, orally administered antibiotics have been increasingly used for serious systemic infections. Antibiotics ideal for oral administration are those that have the appropriate spectrum, high degree of activity against the presumed or known pathogen, and have good bioavailability. Oral antibiotics with high bioavailability, that is > or = 90% absorbed, achieve serum/tissue concentrations comparable to IV administered antibiotics at the same dose. The popularity of "IV to PO switch therapy" is possible because of the availability of many potent oral antibiotics with high bioavailability. Initial IV therapy is appropriate in patients who are in shock/have impaired intestinal absorption, but after clinical defervescence, completion of therapy should be accomplished with oral antibiotics. As experience with "IV to PO switch therapy" has accumulated, confidence in oral antimicrobics for therapy of serious systemic infections has continued to increase. The trend in treating serious systemic infections entirely with oral antimicrobial therapy will continue, and is clearly the wave of the future. PMID- 17116445 TI - Antibiotic drug interactions. AB - Drug-drug interactions in the field of infectious diseases continue to expand as new drugs are approved, metabolic enzymes and transporters are identified, and recommendations for co-administration of drugs are revised. This article provides an overview of the principles and mechanisms of drug-drug interactions and describes pharmacokinetic-pharmacodynamic interactions commonly associated with antibacterial therapy, antiviral agents (non-retroviral), and drugs for tuberculosis. PMID- 17116446 TI - Antibiotic selection in the penicillin-allergic patient. AB - Clinicians should be familiar with which antibiotics are safe to use for different types of penicillin-allergic reactions. Clinically, it is convenient to divide patients with penicillin allergy into three categories: those with unknown or possible reactions to penicillin, those with a drug fever or rash, and those with hives or anaphylactic reactions. Beta-lactam antibiotics may be used safely for patients with unknown/possible penicillin allergy and drug fever or rash. Penicillins or beta-lactams should not be used for patients with hives or anaphylactic reactions. For all patients, clinicians should consider antimicrobial therapy with an antibiotic that does not cross-react with penicillins or beta-lactams. This article reviews how clinicians should select antimicrobials in penicillin-allergic patients. PMID- 17116448 TI - Foreword. PMID- 17116447 TI - Clinical approach to antibiotic failure. AB - A systematic approach is presented for the patient with antibiotic failure. Noninfectious mimics of infection and nontreatable infections must first be excluded. Then, the clinician must identify those patients who have responded but have a surgical component of their infection or have complications separate from their initial infection. Such complications could include drug fever, phlebitis, decubiti, urinary tract infection, aspiration, and pulmonary embolism. Other patients may deteriorate clinically because of incorrect antibiotic coverage, failure of antibiotic to reach the site of infection, local inactivation of antibiotic, paradoxical response, immune compromise, or because they have reached the point of no return. PMID- 17116450 TI - Critical care medicine: landmarks and legends. AB - Critical care medicine was born from the selective pressures of human disease, and with the perseverance and foresight of a select few pioneers, has become an independent field of medicine. This introduction travels back in time to evaluate those visionaries and their landmark contributions. Advancements in caring for the critically ill and organ failure occurred during the wars of the twentieth century. Landmark advances in the management of respiratory paralysis occurred in the polio epidemic of the 1940s. It was during this era that the world's first ICU was developed. Contemporary critical care differs considerably from that which marked its birth. Much of the technology we currently employ is assumed: invasive hemodynamic monitoring, mechanical ventilation, antisepsis, and antibiotics. PMID- 17116451 TI - Management of peritonitis in the critically ill patient. AB - Severe secondary peritonitis carries significant mortality, despite advancements in critical care support and other therapies. Surgical management requires a multidisciplinary approach to guide the timing and the number of interventions necessary to eradicate the septic foci and create optimal healing with the fewest complications. Research is needed regarding the best surgical strategy for very severe cases. The use of deferred primary anastomosis seems safe in patients presenting with hemodynamic instability and hypoperfusion. These patients have a high risk of anastomotic failure and fistula formation. Allowing for aggressive resuscitation and judicious assessment of the progression of local inflammation are safe strategies to achieve the highest success and minimize serious and protracted complications in patients who survive the initial septic insult. PMID- 17116452 TI - The relationship between the surgeon and the intensivist in the surgical intensive care unit. AB - When a patient enters the intensive care unit, the admitting surgeon also enters a new environment. In some hospitals, the surgical intensive care unit (SICU) is "closed"--critical care providers manage care; in others the unit is "open," and the admitting surgeon is in charge. A third system is the "mixed" model of ICU administration; a collaborative approach. This article addresses concerns and conflicts that frequently arise between admitting surgeons and intensivists. It is written from the perspective of two surgeon-intensivists who have been in both roles. Recent behavioral and social research on ICU conflicts and their resolution is reviewed, and and new strategies for conflict resolution are also presented. PMID- 17116454 TI - Basic ventilator management: lung protective strategies. AB - Acute respiratory failure is manifested clinically as a patient with variable degrees of respiratory distress, but characteristically an abnormal arterial blood partial pressure of oxygen or carbon dioxide. The application of mechanical ventilation in this setting can be life-saving. An emerging body of clinical and basic research, however, has highlighted the potential adverse effects of positive pressure ventilation. Clinicians involved with the care of critically ill patients must recognize and seek to prevent these complications using lung protective ventilation strategies. This article discusses the basic concepts of mechanical ventilation, reviews the categories of ventilator-associated lung injury, and discusses current strategies for the recognition and prevention of these adverse effects in the application of mechanical ventilation. PMID- 17116455 TI - Ventilator-associated pneumonia. AB - Ventilator-associated pneumonia (VAP) is a significant clinical infection affecting up to one-third of patients requiring mechanical ventilation, and is associated with significant attributable morbidity and mortality. Clinicians should have a heightened clinical suspicion for VAP with diagnostic goals focusing on accuracy; gathering of lower respiratory tract culture; and appropriate and timely initial antibiotic therapy. Early and adequate antibiotic therapy is important to optimize the management of patients with VAP. The incidence and etiologic patterns of the major pathogens causing VAP must be taken into account when making empiric antibiotic therapy choices. Subsequent de escalation and prescription of an appropriate duration of therapy guided by clinical response and culture results may lead to decreased morbidity and future antibiotic resistance. PMID- 17116453 TI - Critical care issues in the early management of severe trauma. AB - Violent trauma and road traffic injuries kill more than 2.5 million people in the world every year, for a combined mortality of 48 deaths per 100,000 population per year. Most trauma deaths occur at the scene or in the first hour after trauma, with a proportion from 34% to 50% occurring in hospitals. Preventability of trauma deaths has been reported as high as 76% and as low as 1% in mature trauma systems. Critical care errors may occur in a half of hospital trauma deaths, in most of the cases contributing to the death. The most common critical care errors are related to airway and respiratory management, fluid resuscitation, neurotrauma diagnosis and support, and delayed diagnosis of critical lesions. A systematic approach to the trauma patient in the critical care unit would avoid errors and preventable deaths. PMID- 17116456 TI - Practical issues of hemodynamic monitoring at the bedside. AB - The hemodynamic monitoring of a surgical patient acquires a major relevance in high-risk patients and those suffering from surgical diseases associated with hemodynamic instability, such as hemorrhagic or septic shock. This article reviews the fundamental physiologic principles needed to understand hemodynamic monitoring at the bedside. Monitoring defines stability, instability, and response to therapy. The major hemodynamic parameters measured and derived from invasive hemodynamic monitoring, such as arterial, central venous, and pulmonary catheterization, are discussed, as are its clinical indications, benefits, and complications. The current clinical data relevant to hemodynamic monitoring are reviewed and discussed. PMID- 17116457 TI - Management of severe sepsis in the surgical patient. AB - Sepsis and septic shock are not uncommon conditions in the surgical intensive care unit. Sepsis is a generalized activation of the immune system in the presence of clinically suspected or culture-proven infection. Severe sepsis is sepsis with organ system dysfunction. Septic shock is sepsis with hypotension (systolic blood pressure <90 mm Hg) without other causes. Although the incidence of sepsis is increasing, the case fatality rate is falling. This improvement in outcome is in part due to bold initiatives like the Surviving Sepsis Campaign from the Institute for Health Care Improvement. In this article the authors present the epidemiology of severe sepsis and evidence-based campaigns for its treatment, with a focus on the surgical patient. PMID- 17116458 TI - Advances in surgical nutrition. AB - Dr. Stanley Dudrick invented total parenteral nutrition in 1968, providing a desperately needed therapy to those patients who could not eat. It has since saved thousands of patients worldwide. Nutrition interventions (NI) in surgical/trauma and critically ill patients have evolved dramatically during the last 20 years from a supportive therapy to a clear therapeutic role. Like any other form of therapy, NI will benefit patients when adequately indicated and prescribed. NI, however, may cause significant side effects and harm when poorly ordered. This article reviews the indications for the prescription of the different forms of NI available to the clinician caring for the surgical patient. PMID- 17116459 TI - Jaundice in the intensive care unit. AB - Hyperbilirubinemia, or jaundice, is common in the ICU, with incidence up to 40% among critically ill patients. Unfortunately, it is poorly understood in the critically ill, and too often presents a diagnostic dilemma to the ICU physician. Causes of jaundice in the ICU are multiple; the etiology in any given patient, multifactorial. Acute jaundice can be a harbinger or marker of sepsis, multisystem organ failure (MSOF), or a reflection of transient hypotension (shock liver), right-sided heart failure, the metabolic breakdown of red blood cells, or pharmacologic toxicity. Acute ICU jaundice is best divided into obstructive and nonobstructive. This stratification directs subsequent management and therapeutic decisions. PMID- 17116460 TI - Pharmacologic support of the failing heart. AB - Cardiovascular failure in critically ill patients carries a high mortality. Identification and treatment of the underlying etiology simultaneously with prompt therapy are indicated to avoid the consequences of prolonged shock. Physicians should assess patients using all available clinical, radiologic, and laboratory data to avoid the pitfalls associated with use of single measures of regional or global perfusion. Continued evidence of inadequate perfusion despite fluid resuscitation warrants consideration of placement of a pulmonary artery catheter or pharmacologic support of the cardiovascular system. Finally, the dynamic nature of physiology in critically ill patients requires constant patient reassessment and flexibility in treatment to tailor therapy individually as the pathologic state evolves. PMID- 17116461 TI - Management of sepsis. AB - Severe sepsis remains a common cause of death in surgical patients. Eradication of the septic source and supportive care has long been the mainstay of treatment. In recent years, however, early goal-directed therapy, tighter glucose control, administration of drotrecogin alfa (activated), and steroid replacement have produced improved morbidity and mortality. In the future, a better understanding of the pathophysiology of sepsis and clinical studies may further improve outcomes from severe sepsis. PMID- 17116462 TI - Brain death and withdrawal of support. AB - The definition of death has evolved to include the concept of brain death. The brainstem is an indispensable central integrative unit for all vital functions. The clinical criteria for brain death consist of the demonstration of the absence of function of the brainstem. Confirmatory testing, which mostly evaluates higher clinical function, is usually not required for the diagnosis of brain death. PMID- 17116464 TI - Non-radioactive analysis of lipid-linked oligosaccharide compositions by fluorophore-assisted carbohydrate electrophoresis. AB - Lipid-linked oligosaccharides (LLOs) are the donors of glycans that modify newly synthesized proteins in the endoplasmic reticulum (ER) of eukaryotes, resulting in formation of N-linked glycoproteins. The vast majority of LLO analyses have relied on metabolic labeling with radioactive sugar precursors, but these approaches have technical limitations resulting in many important questions about LLO synthesis being left unanswered. Here we describe the application of a facile non-radioactive technique, fluorophore-assisted carbohydrate electrophoresis (FACE), which circumvents these limitations. With FACE, steady-state LLO compositions can be determined quantitatively from cell cultures and animal tissues. We also present FACE methods for analysis of phosphosugars and nucleotide sugars, which are metabolic precursors of LLOs. PMID- 17116465 TI - Identification of N-glycan-binding proteins for E3 ubiquitin ligases. AB - N-glycans serve as a degradation signal by the SCF(Fbx2) ubiquitin ligase complex in the cytosol. Fbx2, an F-box protein, binds specifically to proteins attached with N-linked high-mannose type oligosaccharides, and subsequently contributes to ubiquitination of glycoproteins. Pre-integrin beta1 is identified as one of the Fbx2 targets. These two proteins bind in the cytosol after inhibition of the proteasome. These results indicate that SCF(Fbx2) ubiquitinates N-linked glycoproteins, which are translocated from the endoplasmic reticulum to the cytosol by the quality control mechanism. This chapter describes methods, including a binding protein assay for N-glycans, a ubiquitination assay for N linked glycoproteins with SCF(Fbx2) ubiquitin ligase complex, an overlay assay for the detection of Fbx2 binding proteins, and a pull-down assay for the interaction between Fbx2 and N-linked glycoproteins, used to identify N-glycan binding proteins for E3 ubiquitin ligases. PMID- 17116466 TI - Family 47 alpha-mannosidases in N-glycan processing. AB - Alpha-mannosidases in eukaryotic cells are involved in both glycan biosynthetic reactions and glycan catabolism. Two broad families of enzymes have been identified that cleave terminal mannose linkages from Asn-linked oligosaccharides (Moremen, 2000), including the Class 1 mannosidases (CAZy GH family 47 (Henrissat and Bairoch, 1996)) of the early secretory pathway involved in the processing of N-glycans and quality control and the Class 2 mannosidases (CAZy family GH38 [Henrissat and Bairoch, 1996]) involved in glycoprotein biosynthesis or catabolism. Within the Class 1 family of alpha-mannosidases, three subfamilies of enzymes have been identified (Moremen, 2000). The endoplasmic reticulum (ER) alpha1,2-mannosidase I (ERManI) subfamily acts to cleave a single residue from Asn-linked glycans in the ER. The Golgi alpha-mannosidase I (GolgiManI) subfamily has at least three members in mammalian systems (Herscovics et al., 1994; Lal et al., 1994; Tremblay and Herscovics, 2000) involved in glycan maturation in the Golgi complex to form the Man(5)GlcNAc(2) processing intermediate. The third subfamily of GH47 proteins comprises the ER degradation, enhancing alpha mannosidase-like proteins (EDEM proteins) (Helenius and Aebi, 2004; Hirao et al., 2006; Mast et al., 2005). These proteins have been proposed to accelerate the degradation of misfolded proteins in the lumen of the ER by a lectin function that leads to retrotranslocation to the cytosol and proteasomal degradation. Recent studies have also indicated that ERManI acts as a timer for initiation of glycoprotein degradation via the ubiquitin-proteasome pathway (Hosokawa et al., 2003; Wu et al., 2003). This article discusses methods for analysis of the GH47 alpha-mannosidases, including expression, purification, activity assays, generation of point mutants, and binding studies by surface plasmon resonance. PMID- 17116467 TI - A cytoplasmic peptide: N-glycanase. AB - A cytoplasmic peptide:N-glycanase (PNGase) has been implicated in the proteasomal degradation of aberrant glycoproteins synthesized in the endoplasmic reticulum. The reaction is believed to be important for subsequent proteolysis by the proteasome since bulky N-glycan chains on misfolded glycoproteins may impair their efficient entry into the interior of the cylinder-shaped 20S proteasome, where the active sites of the proteases reside. The deglycosylation reaction by PNGase brings about two major changes on substrate proteins; one is a removal of N-glycan chains, and the other is the introduction of negative charge(s) into the core peptide by converting glycosylated asparagine residue(s) into aspartic acid residue(s). Therefore, PNGase action can be accurately monitored by detecting both changes using two different methods; that is, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for deglycosylation and isoelectric focusing for detection of introduction of negative charge(s) into core proteins. This chapter will describe the simple in vivo as well as in vitro assay method to detect PNGase activity. PMID- 17116468 TI - Glycomic profiling of cells and tissues by mass spectrometry: fingerprinting and sequencing methodologies. AB - Over the past decade, rapid, high-sensitivity mass spectrometric strat-egies have been developed and optimized for screening for the types of N- and O-glycans present in a diverse range of biological material, including secretions, cell lines, tissues, and organs. These glycomic strategies are based on matrix assisted laser desorption/ionization (MALDI) time-of-flight mass fingerprinting of permethylated derivatives, combined with electrospray (ES) or MALDI tandem mass spectrometry (MS/MS) sequencing and gas chromatography (GC)-MS linkage analysis, complemented by chemical and enzymatic degradations. Protocols for these methods are described in the first part of this chapter. Glycomic experiments yield large volumes of MS data, and interpretation of the resulting spectra remains a time-consuming bottleneck in the process. In the second part of this chapter, we describe the use and operation of a mass spectral viewer program capable of displaying and automatically labeling spectra arising from MALDI fingerprinting of N-glycans. PMID- 17116469 TI - Structural analysis of sialyl N-glycan using pyridylamination and chromatography followed by multistage tandem mass spectrometry. AB - Multiple-dimensional mapping (n-DM) methods consisting of pyridylamination and high-performance liquid chromatography (HPLC) are widely used for oligosaccharide analysis. These methods are quantitative, sensitive, and suitable for separating isomers. Oligosaccharide structures are suggested by elution positions on two or three kinds of columns and then confirmed by enzymatic and chemical treatments or mass spectrometry (MS). Multiple-stage tandem MS (MS(n)) analyses have been used to determine oligosaccharide structures by spectrum matching, comparing standard oligosaccharides, and offering detailed MS(n) fragment analysis. However, oligosaccharides usually exist as a mixture including isomers. The 3-DM method provides quantitative information and well-isolated sialyl- or neutral oligosaccharides; on the other hand, the MS(n) method saves time by eliminating complicated enzyme degradation steps. The combination of both methods, which support each other, represents a reasonable and efficient strategy. This chapter describes 3-DM on HPLC after negative-ion MS(n) spectral matching for sialyl N glycans. PMID- 17116470 TI - Determination of glycosylation sites and disulfide bond structures using LC/ESI MS/MS analysis. AB - Significant progress has been made in discovering and cloning a host of proteins, including a range of glycoproteins. The availability of their predicted amino acid sequences provides useful information, including potential N-linked glycosylation sites. However, only a limited number of protein structures have been solved, and very little is known about the structures of membrane proteins. One of the important structural elements of a protein is its disulfide bonds. These covalent bonds place conformational constraints on the overall protein structure, and thus, their identification provides important structural information. A second important posttranslational modification found in proteins is N-linked glycosylation. Although potential sites of N-linked glycosylation can be predicted from a protein's primary sequence based on the presence of N-X-S/T sequences, not all of the predicted sites will be glycosylated. Therefore, N linked glycosylation sites must be located by structural analysis. We have developed a simple and sensitive method for determining the presence of free cysteine (Cys) residues and disulfide-bonded Cys residues, as well as the N linked glycosylation sites in glycoproteins by liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) in combination with protein database searching using the programs Sequest and Mascot. The details of our method are described in this chapter. PMID- 17116471 TI - Identification of O-GlcNAc sites on proteins. AB - O-linked N-acetylglucosamine (O-GlcNAc) is a monosaccharide posttranslational modification that modifies serine/threonine residues of nucleocytoplasmic proteins in metazoans. O-GlcNAc, like phosphorylation, is dynamic and responsive to numerous stimuli in diverse regulatory pathways. O-GlcNAc may also be found adjacent to or at the same sites as phosphorylation, demonstrating the potential for a reciprocal function on some of these proteins. Like most posttranslational modifications, O-GlcNAc is substoichiometric and may be found at multiple sites with other posttranslational modifications present. Additionally, there is no consensus sequence defining the addition of O-GlcNAc to the peptide backbone, further complicating identification and site mapping. This chapter describes several strategies to confirm that proteins are O-GlcNAc modified and provide subsequent determination of O-GlcNAc attachment sites. We have listed the strengths and limitations of each protocol to allow readers to decide which suits their system and availability of resources. These protocols include galactosyltransferase labeling, immunoblotting, using mass spectrometry based on beta-elimination followed by Michael addition with dithiothreitol, and chemoenzymatic labeling, enrichment, and detection. PMID- 17116472 TI - Chemoenzymatic synthesis of glycan libraries. AB - The expanding interest for carbohydrates and glycoconjugates in cell communication has led to an increased demand of these structures for biological studies. Complicated chemical strategies in glycan synthesis are now more frequently replaced by regio- and stereo-specific enzymes. The exploration of microbial resources and improved production of mammalian enzymes have established glycosyltransferases as an efficient complementary tool for glycan synthesis. In this chapter, we demonstrate the feasibility of preparative enzymatic synthesis of different categories of glycans, such as blood group and tumor-associated poly N-acetyllactosamines antigens, ganglio-oligosaccharides, N- and O-glycans. The enzymatic approach has generated over 100 novel oligosaccharides in amounts allowing milligram to gram distribution to many researchers in the field. Our diverse library has also formed the foundation for the successful developments of both the noncovalent enzyme-linked immunosorbent assay glycan array and the covalent printed glycan microarray. PMID- 17116473 TI - Glycoconjugate vaccines against Haemophilus influenzae type b. PMID- 17116474 TI - Assay of human gastric mucin as a natural antibiotic against Helicobacter pylori. AB - Helicobacter pylori infects more than half of the world's population and is considered a leading cause of peptic ulcer and gastric carcinoma. Although a large number of persons are infected with H. pylori, only a limited number of those infected (approximately 3%) develop peptic ulcers and gastric carcinoma. The progression of the disease is restricted by deeper portion of the gastric mucosa, and in many persons glandular atrophy appears to be prevented by mucins secreted in the deeper portion of the mucosa. Recent studies have shown that this inhibitory activity is at least partly due to the expression of alpha1,4-N acetylglucosamine residues attached to the mucin (MUC6) in the deeper portion of the mucosa. alpha1,4-N-acetylglucosamine residues inhibit cholesterol alpha glucosyltransferase, the product of which constitutes a major component of H. pylori cell wall. This inhibitory activity is thus regarded as a natural antibiotic function. This chapter describes the assay for antibiotic activity of MUC6 mucin against H. pylori infection and growth as well as inhibition by alpha1,4-N-acetylglucosamine-capped mucin-type oligosaccharides. PMID- 17116475 TI - Molecular contacts between antibiotics and the 30S ribosomal particle. AB - Crystal structures of complexes between ribosomal particles and antibiotics have pinned down very precisely the discrete binding sites of several classes of antibiotics inhibiting protein synthesis. The crystal structures of complexes between various antibiotics and ribosomal particles show definitively that ribosomal RNAs (rRNAs), rather than ribosomal proteins, are overwhelmingly targeted. The antibiotics are found at messenger RNA or transfer RNA binding sites and, most importantly, at pivot locations that are key for the structural rearrangements during the molecular mechanical steps in initiation, elongation, or termination of protein synthesis. We focus here on the 30S particle. Structurally, the antibiotics interact in many ways with RNA: (i) only with the phosphate groups (streptomycin); (ii) mainly with bases (hygromycin, spectinomycin); (iii) with a mixture of both (paromomycin, Geneticin); (iv) via magnesium ions (tetracycline) or a protein side chain (streptomycin). The antibiotics can mimic base stacking (pactamycin) or form pseudo-base pairing interactions with ribosomal bases (paromomycin and related aminoglycosides). Resistance strategies (mutations or methylations in rRNA or enzymatic modifications of the antibiotics) can generally be understood on the basis of the intermolecular contacts made between the antibiotics and rRNA residues in the crystal structures. In humans, toxicity of ribosomal antibiotics is most likely due, at least in part, to the sensitivity of mitochondrial ribosomes, since mitochondria evolved from a bacterial ancestor. Antibiotic families (e.g., aminoglycosides) form a set of invariant H-bonds to defined rRNA residues. When such residues are conserved in bacteria, but not in eukaryotes, resistance of eukaryotic ribosomes is observed. The structural knowledge, together with comparative genomic analysis, should allow for the development of new broad spectrum antibiotics with higher selectivity toward bacterial ribosomes and less toxicity on eukaryotic cytoplasmic and mitochondrial ribosomes. PMID- 17116476 TI - Mechanism-based inhibitors to probe transitional states of glycoside hydrolases. AB - Recent structural and kinetic studies indicate that glycosidases (glycoside hydrolases) change the peripheral structure of their catalytic sites dynamically to trim glycan structures. Inhibitors that label specific amino acid residues in the active site of these enzymes based on its mechanism of action are powerful tools to probe such a hidden transitional state. This chapter describes methods of mechanism-based irreversible inhibitors having fluorescence tags, including synthesis, inhibitory assay, rapid separation of the peptides containing labeled residues using antibody column, and proteomic analysis of key amino acid residues using matrix-assisted laser desorption/ionization-time-of-flight (TOF)/TOF mass spectrometry. PMID- 17116477 TI - Regulating cell surface glycosylation with a small-molecule switch. AB - Correct localization of Golgi-resident enzymes is essential for the formation of specific glycan epitopes. In this chapter, we describe a method to control the localization, and thus the activity, of an individual glycosyltransferase by administration of a small molecule. Our method takes advantage of the modularity of most Golgi-resident enzymes, which are composed of localization and catalytic domains. These domains can be physically separated and fused to the small molecule binding proteins FRB and FKBP, which dimerize in the presence of rapamycin. In this way, rapamycin serves as a "switch" for enzyme activity. PMID- 17116478 TI - Metabolic labeling of glycans with azido sugars for visualization and glycoproteomics. AB - The staggering complexity of glycans renders their analysis extraordinarily difficult, particularly in living systems. A recently developed technology, termed metabolic oligosaccharide engineering, enables glycan labeling with probes for visualization in cells and living animals, and enrichment of specific glycoconjugate types for proteomic analysis. This technology involves metabolic labeling of glycans with a specifically reactive, abiotic functional group, the azide. Azido sugars are fed to cells and integrated by the glycan biosynthetic machinery into various glycoconjugates. The azido sugars are then covalently tagged, either ex vivo or in vivo, using one of two azide-specific chemistries: the Staudinger ligation, or the strain-promoted [3+2] cycloaddition. These reactions can be used to tag glycans with imaging probes or epitope tags, thus enabling the visualization or enrichment of glycoconjugates. Applications to noninvasive imaging and glycoproteomic analyses are discussed. PMID- 17116479 TI - Functional proteomic profiling of glycan-processing enzymes. AB - Glycoconjugates play critical roles in regulating cellular and organismal functions. Consequently, defining the relative levels of these glycoconjugates under varied physiological conditions is important. Thus identifying and understanding the regulation of the enzymes that process these glycoconjugates are essential steps in understanding the role of this "glycocode" in development and disease. Activity-based affinity reagents are useful tools for probing these enzymes and should facilitate the unraveling of proteomes. One advantage of activity-based affinity probes is that they can simultaneously reveal multiple enzymes having similar activities. These probes can also be used to enrich proteomes of interest, thereby facilitating identification and cloning of new carbohydrate-processing enzymes. Here we review the current state of activity based affinity probes for profiling carbohydrate-processing enzymes, focusing on successes and limitations, general design features, and a specific example describing profiling of exoglycosidases from cell lysates. PMID- 17116480 TI - Oligosaccharide microarrays to map interactions of carbohydrates in biological systems. AB - Carbohydrate microarrays are becoming a standard tool for glycobiologists to screen large numbers of sugars and elucidate the role of carbohydrates in biological systems. This article describes detailed methods to prepare and use microarrays containing synthetic oligosaccharides as well as a summary of the biological information that can be obtained by using this technology. These methods use different linking chemistries to immobilize a wide range of synthetic oligosaccharides onto glass slides through the formation of a covalent bond. Therefore, this technology enables the elaborate study of a great variety of carbohydrate interactions. PMID- 17116481 TI - Identification of ligand specificities for glycan-binding proteins using glycan arrays. AB - Protein-glycan interactions mediate diverse biological processes in cell communication and innate immunity. They involve the binding of a protein on one cell surface to a glycosylated protein or lipid on an opposing cell surface. Understanding the functional significance of these interactions is of major interest to the scientific community. Numerous studies have demonstrated the utility of solid-phase glycan arrays as a means of identifying glycan binding specificity. These approaches share a common format in that glycans are attached in some manner to a solid-phase surface and the glycan-binding protein (GBP) is presented in solution for binding analysis. Multiple options are available to investigators for detecting these interactions, but the most robust reporters are fluorescence based, and binding can be detected as relative fluorescence units. The solid-phase assays have been proved to be highly scalable and suitable for manufacturing processes. The latest innovations have led to the production of miniaturized arrays using microarray printing technology to produce glycan microarrays with the potential to spot and interrogate several thousands of unique glycans simultaneously. This chapter describes two glycan array platforms developed by the Consortium for Functional Glycomics, a microtiter plate-based array and a covalent printed array, and the analytic methods used to conduct binding specificity assays for a broad range of GBPs and organisms. PMID- 17116482 TI - High-throughput analysis of lectin-oligosaccharide interactions by automated frontal affinity chromatography. AB - Frontal affinity chromatography (FAC) is a quantitative method that enables sensitive and reproducible measurements of interactions between lectins and oligosaccharides. The method is suitable even for the measurement of low-affinity interactions and is based on a simple procedure and a clear principle. To achieve high-throughput and efficient analysis, an automated FAC system was developed. The system designated FAC-1 consists of two isocratic pumps, an autosampler, and a couple of miniature columns (bed volume, 31.4 microl) connected in parallel to either a fluorescence or an ultraviolet detector. By use of this parallel-column system, the time required for each analysis was reduced substantially. Under the established conditions, fewer than 10 hrs are required for 100 interaction analyses, consuming as little as 1 pmol pyridylaminated oligosaccharide for each analysis. This strategy for FAC should contribute to the construction of a lectin oligosaccharide interaction database essential for future glycomics. Overall features and practical protocols for interaction analyses using FAC-1 are described. PMID- 17116483 TI - Preparation of neoglycolipids with ring-closed cores via chemoselective oxime ligation for microarray analysis of carbohydrate-protein interactions. AB - Affinities of most oligosaccharide-protein interactions are so low that multivalent forms of ligand and protein are required for detecting interactions. The neoglycolipid (NGL) technology was designed to address the need for microscale presentation of oligosaccharides in a multivalent form for studying carbohydrate-protein interactions, and this is now the basis of a state-of-the art carbohydrate microarray system. NGL technology involves conjugating oligosaccharides by reductive amination to the aminolipid 1,2-dihexadecyl-sn glycero-3-phosphoethanolamine (DHPE). Other than ring-opening of the monosaccharide residues at reducing ends, oligosaccharides remain intact, and the NGLs derived from trisaccharides or larger oligosaccharides have performed well for the majority of carbohydrate-recognition systems that have the peripheral or backbone regions of oligosaccharides as recognition motifs. However, ring-opening of reducing end monosaccharides limits applicability to very short oligosaccharides (di- and trisaccharides) and, potentially, to N-glycans recognized by proteins such as Pisum sativum agglutinin (pea lectin) that require both intact core and backbone regions for strong binding. This chapter describes a method for preparing NGLs (designated AO-NGLs) from reducing oligosaccharides by chemoselective oxime-ligation to a new lipid reagent, N-aminooxyacetyl-DHPE. Microarray analyses of the AO-NGL derived from Lewis x (Le(x)) trisaccharide probed with anti-Le(x) antibodies indicate that a significant proportion of the core monosaccharide linked to lipid is in ring-closed form. Thus, AO-NGLs have broadened the applicability of NGLs as probes in studies of carbohydrate-protein interactions. PMID- 17116484 TI - Development of a lectin microarray based on an evanescent-field fluorescence principle. AB - To investigate protein-carbohydrate interactions in a comprehensive and high throughput manner, carbohydrate biosensors including microarrays have recently attracted increased attention. In this context, carbohydrate and lectin microarrays are emerging as techniques to meet such requisites. However, most of these methods adopt a conventional immuno-detection system, which requires repetitive washing steps before detection. Since lectin-carbohydrate interactions are relatively weak compared with those between antigens and antibodies, a more precise analytical method, which does not require any washing step, is desirable. We describe here a novel platform for lectin microarray that enables direct observation of lectin-carbohydrate interactions under equilibrium conditions, on the basis of an evanescent-field fluorescence-assisted detection principle. This method allows the analysis of a panel of glycoproteins (glycopeptides) in an extremely sensitive manner. The system also allows real-time observation of lectin-glycoprotein interactions in an aqueous phase. No washing procedures are required, thus relatively weak interactions are detectable. The described lectin microarray is expected to be useful for various fields of glycomics requiring high-throughput analysis of not only purified glycoproteins but also of crude samples. PMID- 17116485 TI - Complex secondary chromosome abnormalities in advanced stage anaplastic large cell lymphoma of children and adolescents: a report from CCG-E08. AB - Among pediatric non-Hodgkin lymphomas, one of the most distinctive types is anaplastic large cell lymphoma (ALCL). Specific chromosomal abnormalities are associated with prognosis in childhood acute lymphoblastic leukemia, but chromosome abnormalities have not been evaluated for prognostic value in pediatric ALCL. For Children's Cancer Group protocol CCG-E-08 Etiologic Study of Non-Hodgkin Lymphoma in Childhood, three patients were enrolled with cytogenetic analysis of ALCL and simultaneously enrolled on treatment protocol CCG-552. Pathology material and karyotypes at initial diagnosis underwent central review. Demographics included ages of 9, 12, and 14 years, and a male/female ratio of 1:2. All patients had advanced disease (stage III). Disease progressed or relapsed in two patients, and one died. Chromosomal abnormalities, including t(2;5)(p23;q35), the ALK/NPM fusion gene, and complex karyotypes with multiple additional abnormalities, were identified in all three patients. In two patients with progressive disease or relapse, additional chromosomal abnormalities at 1q21 and 10q24, possibly involving MCL1 and HOX11/TCL3, respectively, may have contributed to worse outcome. Pediatric ALCL cases frequently have complex karyotypes and usually involve ALK/NPM translocations in this limited study. Additional chromosome abnormalities may be involved in the pathogenesis of ALCL. Further studies are warranted in larger cohorts of children and adolescents with ALCL. PMID- 17116486 TI - Chondromyxoid fibroma of the nasal cavity with an interstitial insertion between chromosomes 6 and 19. AB - Chondromyxoid fibroma is an uncommon benign cartilaginous tumor that rarely presents in the sino-nasal region as a locally destructive, erosive lesion. Both clinically and histologically, it is a difficult diagnosis and can be confused with malignant processes such as myxoid chondrosarcoma. Histology of the tumor, especially with a small sample, can be challenging because of its heterogeneous nature showing an admixture of fibrous, myxoid, and chondroid areas. We are reporting unique cytogenetic findings in a case of chondromyxoid fibroma involving the floor of the nasal cavity with a clonal rearrangement between chromosomes 6 and 19. To our knowledge, karyotypes of 14 cases are reported in literature, with 11 cases reporting nonrandom, clonal abnormalities of chromosome 6. These results illustrate the distinctive nature of this tumor and may help identify genes involved in the pathogenesis of this tumor. PMID- 17116487 TI - Intravascular large B-cell lymphoma associated with a near-tetraploid karyotype, rearrangement of BCL6, and a t(11;14)(q13;q32). AB - Chromosome analysis of a patient with intravascular large B-cell lymphoma (IVL) revealed a complex, near-tetraploid karyotype with 83 chromosomes. Abnormalities included a t(11;14)(q13;q32), which was confirmed with both interphase fluorescence in situ hybridization (FISH) using an IGH/cyclin D1 dual-color, dual fusion probe set and cyclin D1 immunohistochemical analysis. Abnormality of 3q was also evident. Interphase FISH analysis with a dual-color, break-apart probe set confirmed rearrangement of BCL6. To our knowledge, this is the first report of these abnormalities in IVL. PMID- 17116488 TI - Alterations in the ATP2A2 gene in correlation with colon and lung cancer. AB - Sarcoendoplasmic reticulum calcium transport ATPases (SERCA-type calcium pumps), proteins that accumulate calcium in the endoplasmic reticulum, play an important role in numerous signaling pathways controlling tumor growth, differentiation, and cell death. Reports that Atp2a2 (Serca2) haploinsufficient mice often developed cancer prompted us to study the involvement of the ATP2A2 gene in human cancer development. We found 13 different novel alterations of the ATP2A2 gene in 27 of 416 alleles of patients with two different types of cancer. Changes in ATP2A2 were significantly more common in patients with colon cancer (P < 0.0001, odds ratio OR = 25.3) or lung cancer (P = 0.046, OR = 8.05). The 13 alterations were missense mutations (2), intronic deletions (2), intronic insertions (1), and single-nucleotide alterations (8: two in the coding region, three in the intronic region, and three in the promoter region). We detected lost or reduced expression of ATP2A2 in all patients with alterations in the promoter region, as well as in patients with a combination of gene alterations. Our results suggest that germline alterations of ATP2A2 may predispose to lung and colon cancer and that an impaired ATP2A2 gene might be involved, directly or indirectly, as an early event in carcinogenesis. PMID- 17116489 TI - Fibrous lipoblastoma with 8q11.2 abnormality. AB - A 3-month-old African American female infant had a rapidly growing lipoblastoma with a prominent fibrous component in the soft tissue of the left lateral knee, which recurred at 10 months. Cytogenetic analysis revealed deletion of 8(q11.2q13) with a 19(q12q13.3) insertion at that site, confirming that this is closely related to the conventional lipoblastoma. The presence of multivacuolated lipoblasts and the staining characteristics (no staining for CD99, CD34, or smooth muscle actin) distinguish this from the recently described lipofibromatosis. PMID- 17116490 TI - A complex translocation (6;12;8)(q25;q24.3;q13) in a fibrous hamartoma of infancy. AB - We report the case of an 18-month-old girl who came to medical attention with a left cervical mass. Surgical excision was performed 16 months later. Histology revealed a fibrous hamartoma of infancy. Follow-up has been uneventful, and no recurrence of the mass has been observed. Cytogenetic analysis showed a complex translocation involving chromosomes 6, 8, and 12, namely, t(6;12;8)(q25;q24.3;q13). This case represents the second cytogenetic analysis reported to date in fibrous hamartoma of infancy and reveals a different translocation than the reciprocal translocation t(2;3)(q31;q21) previously reported. PMID- 17116491 TI - Recurrent genomic imbalances in primary effusion lymphomas. AB - Primary effusion lymphomas (PEL) form a subset of AIDS-related lymphomas and usually have a poor prognosis. Although Kaposi's sarcoma-associated herpes virus (KSHV) is often associated with PEL, very little is known about the exact mechanisms or causative effects of these associations. We investigated the chromosomal imbalances in six KSHV-positive PEL cell lines using comparative genomic hybridization analysis. We defined the shortest regions of overlaps for genomic gains on six chromosomes: 1q31, 4q31 approximately q33, 7q10 approximately q21, 8q21.1, 12q0 approximately q23, and Xp11 approximately q21. The recurrent nature of the gains found in these chromosomal regions suggests that these imbalances play roles in the pathogenesis of PEL. PMID- 17116492 TI - Inversion (11)(p15q22) with NUP98-DDX10 fusion gene in pediatric acute myeloid leukemia. AB - The inv(11)(p15q22), a rare but recurrent chromosome abnormality that creates a NUP98-DDX10 fusion gene, is associated with de novo or secondary myeloid malignancies. We report a case of acute monocytic leukemia presenting this rearrangement, studied using fluorescence in situ hybridization (FISH) and reverse transcriptase-PCR (RT-PCR). We also review the cases of inv(11) associated with NUP98-DDX10 reported in the literature. PMID- 17116493 TI - Cytogenetic and molecular genetic findings in dedifferentiated liposarcoma with neural-like whorling pattern and metaplastic bone formation. AB - Dedifferentiated liposarcoma, a subtype of liposarcoma, is characterized by juxtaposition of well-differentiated liposarcoma with a nonadipocytic sarcoma. A peculiar form of dedifferentiated liposarcoma has been described, characterized by a nonlipogenic component with a neural-like whorling pattern of growth and metaplastic bone formation. We report the cytogenetic and molecular genetic findings of this peculiar form of dedifferentiation in a retroperitoneal tumor found in a 58-year-old woman. The neoplasm had the typical histologic findings and a complex karyotype characterized by several numeric and structural chromosome abnormalities, including the presence of ring and giant rod chromosomes. Molecular genetic studies found high levels of amplification of the MDM2 oncogene, consistent with the amplification of the 12q14 chromosome band, a cytogenetic abnormality commonly found in these tumors. These findings indicate that, despite its unique and peculiar morphologic features, this unusual type of dedifferentiated liposarcoma shares many of the cytogenetic and molecular genetic abnormalities found in other forms of dedifferentiation. However, the specific cytogenetic and molecular determinants of these peculiar morphologic findings remain unknown. PMID- 17116494 TI - Robertsonian translocation (13;14)(q10;q10) in two young male myelodysplastic syndrome patients. PMID- 17116497 TI - Normal sexual response in women. AB - In summary, women's sexual response is characterized as highly variable and influenced by a wide range of determinants, including physiologic, psychosocial, and contextual factors. This complexity is reflected also in the multiple etiologic factors and determinants of sexual problems in women. It is evident in current conceptualizations of normal female sexual response, as presented in this article, in which the circularity and overlap of different components and aspects of sexual response in women are viewed differently from the more linear and invariable trajectory of sexual response in men. It is not surprising, therefore, that treatments that target limited physiologic aspects of women's sexual response, such as PDE-5 inhibitors and other vasoactive agents, have demonstrated little overall effectiveness in treating women's sexual dysfunctions [21, 22]. Rather there is consensus at present regarding the need for a more holistic or biopsychosocial approach to management of sexual dysfunction in women. Clearly the roles of psychologic and interpersonal determinants need to be taken into account in this approach. Perhaps most important and especially relevant for treating female sexual dysfunction is the emphasis on interpersonal factors as contributors to the ultimate sexual satisfaction of women. Clinicians should be especially mindful of this influence when addressing women's sexual problems. Another implication of this research is the need for broader definitions and conceptualization of sexual dysfunction in women. Current diagnostic definitions of female sexual dysfunction are based largely on the traditional sexual response cycle models of Masters and Johnson [7] and Kaplan [9]. With the development of new models and concepts of normal sexual response in women, it is increasingly necessary for new definitions and concepts of sexual dysfunction to be developed. Promising efforts have been made in this regard, although much work remains to be done. PMID- 17116498 TI - Categories of female sexual dysfunction. AB - Female sexual dysfunction is a complex and common problem. Several factors influence female sexual function, including biologic and psychosocial factors. Evaluation of female sexual dysfunction should include a complete medical and psychosocial history and a physical examination. Treatment should be multidisciplinary and depends on the etiology but may include education, psychotherapy or sexual therapy, and in some cases pharmacotherapy. Further research in this area will likely lead to a better understanding of the physiology of female sexuality and to novel therapies for female sexual dysfunction. PMID- 17116499 TI - Taking a sexual history. AB - Why should you inquire about sexual concerns? The importance of sexual health to a woman's quality of life and general health and well-being cannot be overstated, yet the topic is too often ignored by gynecologists and primary care physicians. This article has been geared toward helping overcome some of the barriers to physicians simply opening the door to the topic and conducting at least a basic sexual history and screening of sexual concerns. Although treatment options are addressed elsewhere in this issue, the most effective treatment of all is to ask. You cannot treat a problem if you do not know it exists. PMID- 17116500 TI - Disorders of sexual desire and arousal. AB - Desire and arousal disorders are very common. These disorders can cause significant distress to a patient. A successful approach depends on an accurate diagnosis, which is dependent on history. Laboratory evaluation is usually not helpful, whereas psychosexual therapy is helpful in many cases. Although there is some evidence that drug therapy is helpful in some cases, no drug has been approved for the treatment of these disorders. PMID- 17116501 TI - Dyspareunia. AB - Dyspareunia, better termed women's sexual pain, is a poorly understood disorder once believed to be purely psychologic. Thanks to cooperative research efforts from several specialties toward defining subsets of the disorder, understanding the etiology of subsets and their comorbidities and new concepts for diagnosis and management are being validated or are being put into practice. This review describes the surprising prevalence of sexual pain, outlines new definitions for subtypes of sexual pain and diagnostic criteria for them, and applies these diagnoses to the task of selecting treatment options. PMID- 17116502 TI - Orgasmic dysfunction. AB - Orgasmic disorders are common in women. Unfortunately a lack of consistent, uniform definitions has made this a difficult disorder to study in depth. Etiology is frequently multifactorial, with psychologic issues often playing a prominent role. Diagnosis depends on a detailed history, which then guides treatment to target the underlying causes. Cognitive behavioral therapy has the most favorable outcome evidence to date. PMID- 17116503 TI - Female androgen insufficiency. AB - FAI, seen commonly and often caused by aging and ovarian dysfunction, still presents a challenge to clinicians because of some unresolved and unattended aspects of the condition, including standardized diagnosis and management. Although the use of androgens in women who have FAI has been shown to be effective, there are no FDA approved androgen preparations available at this time for women. Large scale, long-term, controlled trials focusing on establishment of valid and standardized diagnosis and treatment options are needed. PMID- 17116504 TI - Pharmacological effects on sexual function. AB - Many drugs may have effects on sexual function. Sexual function is complex and psychological and relationship issues are likely to have greater impacts on sexual function in women than drugs. Although it is important to understand the effects of drugs on sexual function, physicians should use caution in "medicalization" of sexual function in women [106]. PMID- 17116505 TI - Sexual function after gynecologic cancer. AB - Sexual functioning is the most enduringly compromised quality of life issue women face after treatment gynecologic cancer. As the population of cancer survivors continues to grow, more attention needs to be given to women who have such concerns. It is vital to increase the awareness of sexual health after gynecologic cancer. We must work together with our patients to address and treat sexual concerns. PMID- 17116507 TI - Review of what youth programs do to increase the connectedness of youth with adults. AB - Common sense and psychological research tell us that connections to adults- parents and others--are integral to the process of normal human development. A substantial research literature exists on the role of the parent-child relationship in development, and there is a smaller, but growing body of research that explores the effects of nonparental relationships. Adolescents, in particular, are open to nonparental adults as they strive to create for themselves lives more independent from their parents while still valuing advice from those more experienced than they. The most commonly examined nonparental relationship is that of a teacher and a student. One of the less explored areas of investigation is the importance of relationships youth have with adults they find in their weekend and after-school activities. This article examines field research that has been conducted over the past 15 years on youth programs, to address what has been learned about "connectedness" as it manifests itself in the field. By connectedness, we mean primarily the attachment youth have to the adults in the programs. PMID- 17116508 TI - How do children with eating disorders differ from adolescents with eating disorders at initial evaluation? AB - PURPOSE: To compare the clinical presentation of children with eating disorders (ED) to that of adolescents with ED. METHODS: Demographic, medical, and psychiatric data of all 959 in- and outpatients (85 males, 874 females) 8-19 years of age diagnosed with ED that presented to an academic center between 1997 and 2005 were examined via retrospective record review. Young patients (n = 109) were defined as aged < 13 years at presentation, and older patients (n = 850) > or = 13 years and < 20 years. RESULTS: Compared with older adolescents (mean 15.6 years, SD 1.4), younger patients (mean 11.6 years, SD 1.2) were more likely to be male (chi2 = 9.25, p < .005) or diagnosed with eating disorder not otherwise specified (EDNOS) (chi2 = 5.09, p < or = .05), and less likely to be diagnosed with bulimia nervosa (BN) (chi2 = 13.45, p < or = .001). There were no significant differences in anorexia nervosa (AN) diagnoses between groups. Young patients were less likely to report purging (chi2 = 26.21, p < .001), binge eating (chi2 = 26.53, p < .001), diet pill (chi2 = 13.31, p < .001) or laxative use (chi2 = 6.82, p < .001) when compared with older teens. Young patients weighed less in percentage ideal body weight (p < .05), had a shorter duration of disease (p < .001), and had lost weight more rapidly than older adolescent patients (p < or = .001). CONCLUSIONS: There are important diagnostic and gender differences in younger patients. Young ED patients presented at a lower percentage of ideal body weight and had lost weight more rapidly, which may put them at higher risk for future growth sequelae than their older counterparts. PMID- 17116509 TI - Hospitalization of children and adolescents for eating disorders in the State of New York. AB - PURPOSE: To examine hospitalization patterns, length of stay, cost and insurance status for children and adolescents with Eating Disorders. METHODS: A cross sectional study was conducted of patients with eating disorders aged 9 to 17 years, discharged from hospitals in the State of New York in 1995. All patients discharged in the state were registered by the State Planning and Research Collaborative System (SPARCS). A subset was identified based on coding by the International Classification of Diseases (ICD) for Anorexia Nervosa, Bulimia and Eating Disturbance Not Otherwise Specified. The Statistical Analysis System (SAS) was used for data analysis. The variables selected were gender, ethnicity, insurance status and length of stay. RESULTS: In one year there were 352 hospitalizations, 312 females (88.6%) and 40 males (11.4%); 279 Caucasians (79.3%), 35 African Americans (9.9%), and 38 Other (10.8%); commercial insurance 246 (69.9%), Medicaid 68 (19.3%), other 38 (10.8%). The diagnostic categories were Anorexia Nervosa 242, Bulimia 59, and Eating Disturbance Not Otherwise Specified 63 (reflecting dual diagnosis in 13). Mean length of stay was 18.43 days, the median was 7 days. The cost per stay ranged between 341.78 dollars and 148,471 dollars; with a median of 3817 dollars and a mean of 10,019 dollars. Length of stay was not influenced by gender, age, or ethnicity; only payor status, availability of insurance, was dominant. CONCLUSIONS: This is the first statewide report on hospitalization of children and adolescents for eating disorders. The mean cost in 1995 exceeded 10,000 dollars. A correlation was found between length of stay and insurance status. Hospitalizations for eating disorder have a significant public health impact, calling for the formulation of fair and rational strategies to optimize care. PMID- 17116510 TI - Self-weighing in adolescents: helpful or harmful? Longitudinal associations with body weight changes and disordered eating. AB - PURPOSE: This study addresses the question: Is frequent self-weighing related to changes in body weight and disordered eating behaviors over a 5-year period among adolescent females and males? METHODS: Project EAT is a 5-year population-based longitudinal study. Participants completed surveys exploring factors associated with eating and weight concerns. Participants included 2516 adolescents who were transitioning from early to middle adolescence (younger cohort) and from middle to late adolescence (older cohort). RESULTS: In the older cohort of females and in both cohorts of males, frequent self-weighing at Time 1 was not associated with weight changes at Time 2 (5 years later), after adjusting for Time 1 weight status and sociodemographic characteristics. In the younger cohort of females, Time 1 frequent self-weighing predicted weight increases at Time 2. In both cohorts of females, but not in males, Time 1 frequent self-weighing predicted higher prevalences of Time 2 disordered eating behaviors, including unhealthy weight control behaviors and binge eating, after adjusting for Time 1 behavioral outcomes, weight status, and sociodemographic characteristics. CONCLUSIONS: Frequent self-weighing was not associated with weight change, with the exception of predicting weight increases in younger females. In females, but not males, self-weighing predicted a higher frequency of binge eating and unhealthy weight control behaviors. Although further research is needed to explore the potential merits and problems associated with regular self-weighing, the findings suggest that population-based obesity prevention strategies targeting adolescents should avoid messages likely to lead to frequent self-weighing. PMID- 17116511 TI - Effects of an oral contraceptive (norgestimate/ethinyl estradiol) on bone mineral density in adolescent females with anorexia nervosa: a double-blind, placebo controlled study. AB - PURPOSE: To evaluate the effect of an oral contraceptive (OC) on bone mineral density (BMD) in adolescent females with anorexia nervosa (AN) or eating disorder not otherwise specified (EDNOS). METHODS: Females 11-17 years of age with AN or EDNOS entered the study. Subjects were randomized equally to treatment with a triphasic OC containing norgestimate (NGM) 180-250 microg and ethinyl estradiol (EE) 35 microg or placebo for 13 28-day cycles. Dual energy x-ray absorptiometry scans (DXA) of the lumbosacral spine (LS) and hip were obtained at baseline and after 6 and 13 cycles. RESULTS: Demographic characteristics of the 112 subjects (NGM/EE 53; Placebo 59) who received study drug and had at least one on-treatment DXA were similar between groups for age (mean: 15 years in each group) and body mass index (mean: NGM/EE 17.9 kg/m2; Placebo 17.6 kg/m2). At the end of Cycle 6, there was a significant increase in the mean LS BMD in the NGM/EE group compared with placebo (.020 g/cm2 vs. .008 g/cm2; p = .021); however, at the end of Cycle 13 the mean increase in LS BMD in the NGM/EE group compared with placebo was no longer significant (.026 g/cm2 vs. .019 g/cm2, p = .244). There was no significant difference in change in hip BMD between groups. The incidence of adverse events was similar between groups. CONCLUSIONS: In a group of adolescent females with AN or EDNOS, treatment with a triphasic OC for 13 cycles did not have a statistically significant effect on LS or hip BMD. PMID- 17116512 TI - Effects of nutrition, puberty, and gender on bone ultrasound measurements in adolescents and young adults. AB - PURPOSE: Quantitative ultrasound (QUS) evaluation of bone is attractive for evaluating skeletal status in adolescents, but its use is limited in the United States due to sparse pediatric reference data. This study evaluated associations between radial and tibial speed of sound (SOS) measurements via QUS and demographic, anthropometric and nutritional variables. METHODS: We enrolled 151 healthy participants, aged 11-26 years, during routine visits to an urban adolescent clinic. SOS measurements were obtained using the Omnisense 7000P (Sunlight Medical Ltd., Tel-Aviv, Israel) and correlated with weight, height, gender, race, sexual maturity rating (SMR), and reported nutritional intake. RESULTS: The sample was 53% female; aged 17+/- 2.8 years (mean +/- SD); and 48% African-American, 21% Hispanic, and 21% Caucasian. Seventy percent of males and 91% of females had achieved SMR 5; 96% of females were postmenarchal. Males met the recommended daily allowance for calcium intake, on average; the females did not. Both the girls and boys reported consumption of inadequate vitamin D. Intake of neither calcium nor vitamin D was correlated with SOS. Radial and tibial SOS were significantly higher in those with SMR 5 (p < .001) and were moderately correlated with age in both genders (r = .42-.64, p < .001). In multivariate analyses, age was associated with SOS at both sites (p < .0001). CONCLUSIONS: This study provides QUS measurements of the peripheral skeleton among healthy adolescents. QUS measurements followed similar age and pubertal distributions to dual-energy X-ray absorptiometry (DXA) bone density measurements; other variables did not follow expected trends. Further research is needed to clarify what skeletal properties are assessed by this technique. This study adds to accumulating evidence that many adolescents do not consume adequate vitamin D or calcium. PMID- 17116513 TI - Bone changes in adolescent girls with anorexia nervosa. AB - PURPOSE: To clarify the effect of chronological age, height, lean tissue mass, and menstrual status on standard reported DXA (dual-energy x-ray absorptiometry) measures. METHODS: Growth and DXA data from a retrospective longitudinal cohort of 30 adolescent females with anorexia nervosa (AN) were examined. Areal bone mineral density (aBMD), total body bone mineral content (BMC) and total body bone area were measured and standardized for age, height, lean tissue mass, and total bone area. We then examined the changes in these parameters after 12 months of multidisciplinary treatment. RESULTS: The subjects had lower BMC and aBMD than the age- and height-matched reference population at baseline, there were further decreases in these parameters with treatment. BMC adjusted for lean tissue mass and bone area were normal at baseline and there was no significant change with treatment. Bone area adjusted for height was low at baseline and decreased despite treatment. CONCLUSION: Our data suggest that the low age- and height adjusted bone mineral content and bone mineral density in females with AN is secondary to poor bone growth leading to low bone area. These subjects had relatively normal bone mineral content for their bone size and lean tissue mass. Changes in bone size were proportional to changes in lean tissue mass. PMID- 17116514 TI - Fast food consumption and breakfast skipping: predictors of weight gain from adolescence to adulthood in a nationally representative sample. AB - PURPOSE: To investigate whether fast food consumption and breakfast skipping are associated with weight gain during the transition from adolescence to adulthood. METHODS: A prospective study of 9919 adolescents participating in Waves II (age range 11-21 years) and III (age range 18-27 years) of the National Longitudinal Study of Adolescent Health. BMI z scores (zBMI) were computed using the 2000 Centers for Disease Control and Prevention growth charts. Multivariate regression models assessed the relationship between Wave II fast food and breakfast consumption and change in fast food and breakfast consumption between Waves II and III and weight gain during the transition to adulthood. RESULTS: Marked increases in fast food consumption and decreases in breakfast consumption occurred over the 5-year interval. Greater days of fast food consumption at Wave II predicted increased zBMI at Wave III. Fewer days of breakfast consumption at Wave II and decreases in breakfast consumption between Waves II and III predicted increased zBMI at Wave III. CONCLUSIONS: Fast food consumption and breakfast skipping increased during the transition to adulthood, and both dietary behaviors are associated with increased weight gain from adolescence to adulthood. These behaviors may be appropriate targets for intervention during this important transition. PMID- 17116515 TI - Physician perspectives to inform a new recommendation for meningococcal conjugate vaccine (MCV4). AB - PURPOSE: In January 2005, the U.S. Food and Drug Administration licensed a new tetravalent meningococcal conjugate vaccine (MCV4). Before any policy decisions by the U.S. Advisory Committee on Immunization Practices (ACIP) related to MCV4, the Centers for Disease Control and Prevention requested a study to explore the perspective of primary care physicians regarding different recommendation scenarios for use of MCV4. METHODS: Cross-sectional mail survey of a national random sample of pediatricians (PDs) and family physicians (FPs), conducted January 2005. Respondents chose from four MCV4 recommendation scenarios in terms of ability to implement, perceived patient/parent preferences, scientific evidence, and overall best fit. RESULTS: Response rate to the single-mailing survey was 57%. In terms of ability to implement, respondents generally preferred an MCV4 recommendation targeted to middle-school entry (11-12 years old) or with the Td booster at any age, but on the basis of scientific evidence they favored MCV4 at high school completion. For "overall best fit," relatively equal proportions of respondents favored a recommendation at middle school entry and one linked to Td booster administration (whenever it occurred); there were no significant differences between PDs and FPs. Major influences on willingness to recommend MCV4 were vaccine safety/side effects and insurance coverage/reimbursement. CONCLUSIONS: Support for an MCV4 recommendation at middle school entry is common but not universal among primary care providers. Data suggest that respondents appreciate the potential discrepancy between practical aspects of vaccine delivery and the need to protect those adolescents at greatest risk of disease. Respondents' preferences for the overall best fit appear to prioritize ease of implementation over epidemiologic patterns. PMID- 17116516 TI - Reliability of the 2005 middle school Youth Risk Behavior Survey. AB - PURPOSE: To evaluate the reliability of the middle school version of the 2005 Youth Risk Behavior Survey (MSYRBS) questionnaire. METHODS: A convenience sample of 232 Midwestern seventh and eighth grade middle school students completed the MSYRBS questionnaire twice in a 2-week period (14 days apart). The MSYRBS questionnaire, which queries a variety of health risk behaviors, was administered in a manner that preserved anonymity but allowed Time 1 and Time 2 matching. This was accomplished by using two questionnaire scantrons coded with the same unique number, and destroying all used materials to ensure that each participant was matched with their code. Kappa statistics were calculated for individual questions and group characteristics using SAS. RESULTS: The mean kappa was 62.6% and the median was 66.5%. Kappa statistics for each item ranged from -2.4% (injection drug use) to 83.8% (suicide contemplation). Negative kappa values were found for two items that had extremely small cell sizes. Kappas did not differ by gender, grade, or race. Based on nonoverlapping confidence intervals, there were no items that had significantly different prevalence estimates at Time 1 vs. Time 2. Nine items (24.3%) and one category (alcohol-drugs) had kappas below 61.0%. CONCLUSIONS: This preliminary study suggests that the reliability of the MSYRBS is high over time. A number of items should be further examined to determine whether they should be amended or omitted from future versions of the MSYRBS. Further research with larger and more diverse samples is recommended, potentially involving children as content experts. PMID- 17116517 TI - Changes in adolescents' sources of cigarettes. AB - PURPOSE: No previous research has tracked changes in teen sources for tobacco. Such information might help public health officials to target tobacco control efforts more precisely. This investigation used a two-year longitudinal design to determine (1) how adolescents' sources change and (2) whether the timing of smoking onset and duration of tobacco use predict the number and types of sources accessed. METHODS: A survey assessing usual sources of cigarettes and related variables was administered to 4461 seventh-graders annually. Of the target population, 79% provided baseline data, and 64.2% participated in all surveys. RESULTS: At baseline, 30% of the 1144 smokers got cigarettes from peers, compared with 11% using stores, 6% using vending machines, and 17% who stole them. Age of smoking onset did not predict the number or types of sources teens accessed. We did, however, find a significant effect of duration of smoking, showing that more practiced smokers were more likely to get cigarettes both from stores and from their friends. Further, the longer students smoked, the more likely they were to have friends who smoked. CONCLUSIONS: Our results indicated that the means through which teens got their first cigarettes were similar, regardless of when smoking onset occurred. In contrast, as teens became more established smokers, they increased the number of sources they used and relied more on both stores and peers. Once adolescents become smokers, they form a social network of fellow smokers who support their habit, making it even more difficult to quit. PMID- 17116518 TI - Avoiding "truth": tobacco industry promotion of life skills training. AB - PURPOSE: To understand why and how two tobacco companies have been promoting the Life Skills Training program (LST), a school-based drug prevention program recommended by the Centers for Disease Control and Prevention to reduce youth smoking. METHODS: We analyzed internal tobacco industry documents available online as of October 2005. Initial searches were conducted using the keywords "life skills training," "LST," and "positive youth development." RESULTS: Tobacco industry documents reveal that since 1999, Philip Morris (PM) and Brown and Williamson (B&W) have worked to promote LST and to disseminate the LST program into schools across the country. As part of their effort, the companies hired a public relations firm to promote LST and a separate firm to evaluate the program. The evaluation conducted for the two companies did not show that LST was effective at reducing smoking after the first or second year of implementing the program. Even so, the tobacco companies continued to award grants to schools for the program. PM and B&W's role in promoting LST is part of a public relations strategy to shift the "youth smoking paradigm" away from programs that highlight the tobacco industry's behavior and toward programs in which the industry can be a partner. CONCLUSIONS: Individuals and organizations responsible for developing and implementing tobacco control and youth smoking prevention programs should be aware of PM and B&W's role and motivations to encourage the wide-spread adoption of LST in schools. PMID- 17116519 TI - It takes two: reducing adolescent smoking uptake through sustainable adolescent adult partnership. AB - PURPOSE: To assess the effects of a long-term intervention for tobacco use prevention that targets adolescents (Tobacco Free Duo). METHODS: School-based community intervention combined with repeated cross-sectional surveys over 7 years. The intervention was performed in the County of Vasterbotten, Sweden, where survey data were collected in grade seven to nine schools on an annual basis for 7 years. Data for comparison were collected in grade nine on the national level in Sweden. In the intervention area, the annual number of seventh to ninth grade students participating in the study varied between 1300 and 1650. In the reference data, the number of participating ninth grade students approximated 4500 annually. RESULTS: A significant decrease of nearly 50% was seen in smoking prevalence in the intervention area. The decrease was evident in grades eight and nine (ages 14-15 years) in both boys and girls. At the start of the intervention, smoking prevalence in grade nine was 16.1% in the intervention area and 23% in the national reference group. Although the prevalence in the national sample remained stable, there was a decrease to 9.0% in the intervention area at the end of the study period. CONCLUSIONS: These results suggest that the Tobacco Free Duo program contributed to a reduction in adolescent smoking among both boys and girls. Using a multi-faceted intervention that includes adolescent adult partnership can reduce adolescent smoking uptake, and the intervention has been proven to be sustainable within communities. PMID- 17116520 TI - Cervicitis: to treat or not to treat? The role of patient preferences and decision analysis. AB - PURPOSE: Mucopurulent cervicitis is neither a sensitive nor a specific indicator of antibiotic sensitive infection. This analysis examines the positive and negative ramifications of treating cervicitis empirically as a Chlamydial (CT) infection. It begins where prior analyses leave off, with the number of cases of pelvic inflammatory disease (PID) prevented. METHODS: Three treatments were compared: 1) treat empirically/refer partner; 2) test, treat, and base partner treatment on results; 3) test, base treatment on results. The outcomes were the physical sequelae of PID and the psychological sequelae of being diagnosed with CT in a hypothetical cohort of 500 teenagers with cervicitis, among whom the prevalence of CT averaged 33%, but ranged between 10% and 70%. RESULTS: At a CT prevalence of 33%, Treatments 1 and 2 prevented three times as many cases of PID related physical sequelae (n = 14) as Treatment 3 (n = 5). However, to prevent these 14 cases of physical sequelae, with Treatment 1, 163 teens needlessly suffer the psychological sequelae of a false CT diagnosis and with Treatment 2, 101 do so. The ratio of physical sequelae prevented to psychological sequelae caused, changed in relationship to the prevalence of CT, but was always numerically most favorable with Treatment 3. Moreover, it was the only therapeutic approach for which overall morbidity never exceeded the PID-related physical morbidity incurred in the absence of treatment. CONCLUSIONS: By including the effects of over diagnosing and treating CT, we have demonstrated how the risks and benefits of empiric and nonempiric cervicitis therapy vary in relationship to CT prevalence. Failure to consider both the physical and the psychological aspects of patient well-being may mean that well-intentioned policies to reduce physical morbidity do not result in an overall improvement in health of teenagers. PMID- 17116521 TI - Sexual communication and contraceptive use in adolescent dating couples. AB - PURPOSE: This study had two aims: first, to examine the relationship between general sexual communication and contraceptive use in sexually active adolescent romantic couples, and second, to explore predictors of open communication from characteristics of adolescent couples and individual adolescents. METHODS: Data were drawn from 209 couples dating a minimum of four weeks who participated in the Study of Tennessee Adolescent Romantic Relationships. Seventy-three adolescent dating couples (ages 14-21 years) that engaged in sexual intercourse and completed a sexual communication questionnaire were included in current analyses. RESULTS: Nearly 30% of couples failed to use contraception at first intercourse and almost half of couples did not use contraception every time they had sex. More open communication about sex from both male and female partners was associated with increased contraceptive use. Additionally, adolescents who were more satisfied in their relationships reported more open communication about sex, and adolescent females who self-silenced reported less open communication about sex. Finally, mediation analyses revealed that boys' and girls' relationship satisfaction and girls' self-silencing indirectly predicted contraceptive use through their effects on general sexual communication. CONCLUSIONS: This is the first known study to address individual and dyadic components of sexual communication using reports from both members of established adolescent dating couples. Findings suggest that open sexual communication between intimate partners is important to sexual decision-making. The clinical implications of these findings are discussed. PMID- 17116522 TI - Evaluation of a school-based HIV/AIDS educational intervention in Ukraine. AB - PURPOSE: To assess changes observed in knowledge, attitudes toward people living with human immunodeficiency virus (HIV), self-efficacy and behavioral intentions of senior secondary school students after an HIV/AIDS (acquired immune deficiency syndrome) educational intervention in Vinnitsa, Ukraine. METHODS: A quasi experimental, nonequivalent control group design with pretest-posttest components was employed. Two secondary schools (intervention and control) were randomly assigned to the study. A random sample of the 15-16-year-old students (100 participants in each school) completed anonymous self-reported pre- and postintervention questionnaires. In addition, a three-month follow-up questionnaire was administrated in both schools to assess the longevity of the intervention outcomes. Outcome scale scores were compared between the two groups on baseline, first, and second posttests separately. We used the Pearson chi square test for categorical data and a t-test for difference in mean scores. RESULTS: After intervention we observed significantly higher knowledge, attitudes, and self-efficacy scores among students in the intervention school than in the control school (96.2 +/- 6.2 [SD] vs. 82.6 +/- 8.3, respectively, p < .01; 24.4 +/- 4.4 vs. 21.7 +/- 4.8, respectively, p < .01; 21.9 +/- 2.6 vs. 20 +/ 3.8, respectively, p < .01). As a result of the intervention, we also report significantly higher proportions of students in the intervention school than in the control school who would not intend to use alcohol and narcotics and who intend to use condoms (p < .01). CONCLUSIONS: Our AIDS education program was able to considerably improve students' knowledge, attitudes and self-efficacy, and promoted positive changes in participants' behavior intentions; it should therefore be extended to more schools to multiply its effects. However, in the context of evaluation, studies with actual HIV-risk behavior outcomes are urgent for Ukraine. PMID- 17116523 TI - Television viewing and forms of bullying among adolescents from eight countries. AB - PURPOSE: Based on theories suggesting that frequent television viewers act and react in hostile, malicious, malevolent, or verbally aggressive ways rather than being physically violent, the present study investigates relationships between television viewing and different forms of bullying. METHODS: Multilevel regression models were estimated based on cross-sectional data from 31,177 adolescents aged 11, 13, and 15 years from Canada, Estonia, Israel, Latvia, Macedonia, Poland, Portugal, and the United States who participated in the 2001 2002 Health Behavior in School-aged Children Survey. RESULTS: Although all different forms of bullying were associated with television viewing in bivariate analyses, only the verbal forms (i.e. "calling mean names" and "spreading rumors") remained significant in multiple regression models. These relationships were observed consistently in all eight participating countries. However, the association between television viewing and physical forms of bullying such as kicking, pushing, or shoving around, varied across countries. In most weekend TV viewing cultures, frequent television viewers were prone to kick or push another student in addition to verbal forms of bullying, which was not the case in weekday viewing cultures. CONCLUSIONS: These results demonstrate the importance of limiting adolescents' time engaged in unsupervised television watching, and the need to motivate adolescents to engage in joint family activities or organized after-school activities. PMID- 17116524 TI - The role of public health agencies in providing access to adolescent drug treatment services. AB - PURPOSE: To examine the role of public health agencies (PHAs) in providing access to drug treatment services for adolescents by describing the proportion of youth who obtain access to these services through PHA involvement in school health clinics, juvenile drug courts, and other community agencies. METHODS: Analysis of cross-sectional telephone interview data collected from 1999-2003 from a national sample of 1793 PHA key informants from communities surrounding schools in the nationally representative Monitoring the Future (MTF) study of 8th, 10th and 12th grade students. RESULTS: Fifty-eight percent of youth in the MTF sample were served by PHAs that participated in some way in school health clinics, with 30% served by PHAs that provided resources for drug treatment in schools. Twenty-nine percent of youth were served by PHAs involved in juvenile drug court (JDC) programs, 23% by PHAs acting as JDC referral agencies, and 13% by PHAs providing direct JDC drug assessment, treatment and monitoring services. In addition, 44% of youth were served by PHAs providing drug treatment resources in community settings. Treatment access for youth through PHAs varied by region, race/ethnicity, urbanicity, community income level, and youth population density. The largest variation occurred in access via JDC programs. CONCLUSIONS: PHAs may help bridge gaps between drug treatment need and service provision for adolescents who need access to drug treatment services. Strengthening the linkages between PHAs and schools, juvenile drug courts, and other community settings may serve to increase youth access to drug treatment. PMID- 17116525 TI - Socio-demographic and cultural comparison of overweight and obesity risk and prevalence in adolescents in Southern California and Wuhan, China. AB - PURPOSE: This study performed parallel analyses on two large samples of seventh graders living in Los Angeles, California and in Wuhan, China to make direct comparisons of overweight and obesity risks in Western and Eastern cultural environments. METHODS: Two representative samples of 1772 and 1896 seventh grade students were randomly selected from the public or parochial middle schools in the greater Los Angeles area of Southern California in the United States, and public schools in Wuhan city of China. Two body mass index (BMI) references recently established by the U.S. National Center for Health Statistics (NCHS) and the International Obesity Task Force (IOTF), respectively, were used to define overweight (85th < or = BMI < 95th percentile) and obesity (BMI > or = 95th percentile). Logistic regressions were conducted to examine relationships between socioeconomic status (SES) and obesity risk. RESULTS: The prevalence rates of combined overweight and obesity in Los Angeles and in Wuhan were 43.1% and 12.1%, respectively, by the US-NCHS reference, and 45.8% and 11.9%, respectively, by the IOTF reference. Chinese-American adolescents had higher prevalence rates for overweight and obesity than those from China. SES was positively related to the risk of overweight and obesity in the Chinese sample, whereas a negative association was found in Southern California adolescents. Urbanicity was significantly positively related to higher overweight and obesity prevalence. CONCLUSION: Different overweight and obesity prevalence estimates and SES effects were observed for American and Chinese adolescent samples. Research on the underlying mechanisms is needed to help us to set up a tailored program for obesity prevention in Eastern and Western cultural environments. PMID- 17116526 TI - Effectiveness of an HIV prevention program for secondary school students in Mongolia. AB - PURPOSE: To evaluate the effectiveness of a 3-year human immunodeficiency virus (HIV) prevention program for adolescents attending secondary school in Mongolia. METHODS: Comparisons of knowledge, attitudes, self-efficacy and safe sex practices of grade 10 students from schools with a peer education prevention program to grade 10 students from schools without the intervention. Peer education programs were launched in 2000 across Mongolia. In 2004, survey data was collected among 720 randomly selected students from eight schools with the peer education prevention program and compared with those of 647 students from eight schools without this intervention. Data was collected in Ulaanbaatar and three Mongolian provinces and analyzed using multilevel regression methods. RESULTS: Students of schools with the program were statistically significantly more knowledgeable, had less traditional attitudes, and had greater awareness of their self-efficacy in regards to HIV and sexual health. Students from schools with the peer education program were more likely to practice safe sex, though the difference was not statistically significant. However, safe sex practice was found to be statistically significantly safer in a subset of schools that had small teams of peer educators. CONCLUSION: Adolescents in Mongolia are sexually active and at risk for infection with HIV and other STIs. Peer education programs, particularly those that are managed by small teams, appear effective and should be implemented more broadly. PMID- 17116527 TI - Adolescent predictors of emerging adult sexual patterns. AB - PURPOSE: This study estimates the percentages of young adults who fall into three groups based on the context of sexual transition: (1) those who had vaginal intercourse before marriage (Premaritals), (2) those who postponed sex until after marriage (Postponers), and (3) those who have never had vaginal intercourse (Virgins). The second purpose is to determine adolescent biopsychosocial factors that predict membership in these adult groups. METHODS: Analyses are based on 11,407 respondents ages 18-27 years who participated in Waves I and III of the National Longitudinal Study of Adolescent Health. Adolescent indicators reflecting sociodemographic, biosocial, experiential, and contextual factors were used to predict young adult sexual status using multinomial logistic regression models. RESULTS: About 8% of the sample were virgins and 2% were virgins until marriage. Almost 90% had sex before marriage (Premaritals--referent group). Most predictors of status were similar for males and females. Compared with Premaritals, Virgins were younger, non-Black, not advanced in physical maturity relative to peers in adolescence, had higher body mass indexes, were more religious, and perceived parental disapproval of sex during adolescence. Postponers were also more religious than Premaritals but were older. Female Postponers were non-Black and perceived parental disapproval of sex during adolescence. Male Postponers were less likely to have same-gender attractions or no sexual attractions. CONCLUSIONS: Findings document premarital sexual activity as the almost universal sexual trajectory into young adulthood for these cohorts and underscore the roles of biosocial factors and conventional institutions in emerging sexual patterns. PMID- 17116528 TI - Continuity of care in the ambulatory treatment of adolescents with asthma. AB - PURPOSE: To estimate the level of continuity of care, an important component in the management of asthma, using a general index based on medical visits and a disease-specific index based on prescribed medications, and to compare how the indexes predict the use of health care services among adolescents with asthma. METHODS: A cohort of 5586 adolescents with asthma was selected from the administrative database of the province of Quebec, Canada between 1997 and 1999. Patients were followed for at least 1 year to a maximum of 3.5 years. The main exposure variables were an index of continuity of care based on the number of physicians prescribing a medication for asthma (RxICC) and an index of continuity of care based on the number of unreferred physicians (COC). RESULTS: The average RxICC was .42 +/- .29 and the average COC was .26 +/- .22. The adjusted rate ratios for RxICC as a predictor of hospitalizations and emergency department (ED) visits for asthma were, respectively, .83 (95% confidence interval [CI] .80-.87) and .81 (95% CI .80-.83). No significant association was found between COC, hospitalizations, and ED visits for asthma. CONCLUSION: Continuity of care is low among adolescents with asthma in Quebec. The RxICC was, on average, higher than the COC, because patients were more likely to visit the same physician to treat their asthma than they would for other types of care. Only the RxICC was associated with the risk of hospitalization and ED visits for asthma. PMID- 17116529 TI - Evaluating risk-taking behaviors of youth in military families. AB - This study assessed risk-taking behavior in a unique adolescent population: family members of active and retired military personnel. Significantly fewer adolescents in our group engaged in at-risk behaviors compared with national statistics, demonstrating the need for further research. PMID- 17116530 TI - Risk-taking among adolescents who say they can get a handgun. AB - In a survey of 5657 community-residing adolescents, those who said they could get a handgun in two days reported similar and those who said they could not get a handgun reported fewer risk behaviors and less violence perpetration and victimization than adolescents who have their own handgun. Adolescents who said they could get a handgun reported more exposure to and experience with firearms than those who said they could not get a handgun. Risk reduction efforts regarding adolescents should be expanded to include adolescents who say they can get a handgun. PMID- 17116531 TI - Body art among minority high school athletes: prevalence, interest and satisfaction; parental knowledge and consent. AB - Of 997 minority high school students, 8.6% had tattoos and 8% piercings (excluding earlobes); 21% with tattoos and 59% with piercings would not repeat the experience. Fifty-eight percent with tattoos and 43% with piercings reported parental knowledge before the procedure; 75% and 80%, respectively, were asked for proof of parental consent. PMID- 17116532 TI - Anorexia in a 14-year-old girl: why won't she eat? AB - This is a clinical presentation of a 14-year-old female who had weight loss and vomiting following spinal fusion surgery. Her case was complicated by social and behavioral issues. After an initial diagnosis of atypical eating disorder, an upper gastrointestinal study revealed superior mesenteric artery syndrome. Her course continued to be complicated until consistent follow-up with a nutritionist was established. PMID- 17116534 TI - Re: Seeking to understand case management in New York. PMID- 17116535 TI - Three perspectives on suffering. PMID- 17116536 TI - The Americans with Disabilities Act, 16 years later. PMID- 17116537 TI - How do I find a job in case management? PMID- 17116538 TI - Managing alcohol problems, Part 2. PMID- 17116539 TI - Promoting patient safety: one company's example. AB - Patient safety has become a key issue in health care since the Institute of Medicine (IOM) released the report To Err Is Human: Building a Safer Health System. This landmark report examined the extent of preventable patient injuries and deaths occurring in US hospitals. It was estimated that between 44,000 and 98,000 people die annually as a result of medical errors and that nearly half were preventable. Subsequent studies suggest that the medical error rate is even higher. These statistics are a call to action for case managers to explore creative ways to implement patient safety practices in their systems and procedures. PMID- 17116540 TI - Transforming the traditional discharge planning manual. AB - Discharge planning resources change frequently, and keeping them current, available, and accessible to case management (CM) staff and others is a challenge. Given the time-consuming and labor-intensive nature of updating, many organizations revise them in preparation for the tri-annual survey or when they are hopelessly out-of-date. The reality is that CM staff and others involved with discharges have their own personal "little black book" of resources. Those personal reference tools are treated with reverence and guarded like the family Bible. Organizations are challenged to have up-to-date resources available and accessible for staff in all areas. One approach is to transform the traditional paper-based manual into an electronic document and make it available on the organization's intranet. This approach means that each page of the current paper based manual must be in electronic format, ending the use of overcopied resources that become difficult to read and "grandfathered in" to each successive edition of the paper manual. This option permits easy and rapid updates when resources and information change. PMID- 17116545 TI - Racial disparities in late survival after rectal cancer surgery. AB - BACKGROUND: African-American patients experience higher mortality than Caucasian patients after surgery for most common cancer types. Whether longterm survival after rectal cancer surgery varies by race is less clear. STUDY DESIGN: Using 1992 to 2003 Surveillance, Epidemiology, and End Results (SEER)-Medicare data, we examined race and longterm survival among African-American and Caucasian rectal cancer patients undergoing resection. We identified racial differences in patient characteristics, structure, and processes of care. We then assessed mortality using a Cox proportional hazards model, sequentially adding variables to explore the extent to which they attenuated the association between race and mortality. RESULTS: African-American patients had a substantially poorer overall survival rate than Caucasian patients did. Five-year survival rates were 41% and 50%, respectively (p < 0.0001). African Americans were younger (p=0.006), more likely to reside in low income areas (p < 0.0001), and had more baseline comorbid disease (p < 0.0001). They were also more likely to be diagnosed emergently (p < 0.001) and with more advanced cancer (p < 0.001). Accounting for demographic and clinical characteristics reduced the mortality difference, although it remained pronounced (hazard ratio=1.13, CI=1.01 to 1.26). African Americans were more likely to be treated by low volume surgeons and less likely to receive adjuvant therapy (48.6% versus 60.9%, p < 0.0001). After adjusting for provider variables, the hazard ratio for mortality by race was additionally attenuated and became statistically nonsignificant (hazard ratio=1.05, CI=0.92 to 1.20). CONCLUSIONS: Poorer longterm survival after rectal cancer surgery among African Americans is explained by measurable differences in processes of care and patient characteristics. These data suggest that outcomes disparities could be reduced by strategies targeting earlier diagnosis and increasing adjuvant therapy use among African-American patients. PMID- 17116546 TI - Quality initiative in rectal cancer strategy: a qualitative study of participating surgeons. AB - BACKGROUND: The Quality Initiative in Rectal Cancer (QIRC) Strategy randomized 16 hospitals across Ontario to the QIRC strategy versus minimal intervention. The strategy included a workshop, recruitment of a local opinion leader (OL), operative demonstrations, postoperative checklists, audit, and feedback. Surgeons at each intervention hospital used a standardized approach to select a local OL from their ranks. We assessed the experiences of OL and non-OL surgeons in the QIRC strategy. STUDY DESIGN: Semi-structured qualitative telephone interviews were completed with 8 OLs and 8 non-OL surgeons. Interviews were guided by Grounded Theory. Initial interviews were assessed to ensure that domains of interest were fully explored. Two investigators (FW, MF) independently reviewed all final transcripts and identified themes. Consensus among all investigators was achieved for final themes. RESULTS: All approached surgeons participated in interviews. Seven themes were identified: surgical OLs with subspecialist expertise were supported; surgical OL requires technical expertise; limited role for local OL on changing physician practice in rectal cancer operations; limited effect of identifying local OL on local interactions; operative demonstrations supported; characteristics of operative demonstrator were important; and perceived positive effect of the QIRC strategy on practice. CONCLUSIONS: The experience of local OLs in this trial appears to be mixed. Although some of the themes support the concept of a local OL, other themes suggest the local OL had a limited role and effect. The overall QIRC strategy, and in particular the operative demonstration, was viewed positively and was perceived to have a positive longterm effect on participants' practice. PMID- 17116547 TI - Timely administration of prophylactic antibiotics for major surgical procedures. AB - BACKGROUND: Prophylactic antibiotics (PA) given within 60 minutes before surgical incision decrease risk of subsequent surgical site infection. Nationwide quality improvement initiatives have focused on improving the proportion of patients who receive timely prophylactic antibiotics. STUDY DESIGN: This is a cohort study of major surgical procedures performed in 108 Veterans Affairs hospitals between January and December 2005. Using data from the External Peer Review Program and the National Surgical Quality Improvement Program, we examined factors associated with timely PA administration. Univariate and multivariable analyses were performed. RESULTS: There were 8,137 major surgical procedures: cardiac (2,664), hip and knee arthroplasty (3,603), colon (1,142), arterial vascular (606), and hysterectomy (122). Timely PA occurred in 76.2% of patients, 18.2% received them too early, and 5.4% received them too late. Early administration accounted for 79% of untimely PA. Differences in timeliness were seen by procedure type (68% to 87%; p < 0.0001), admission status (67% to 80%; p < 0.0001), and antibiotic class (65% to 89%; p < 0.0001). PA administration occurred in the operating room for 63.5% of patients. When PA administration occurred in the operating room, they were timely in 89% of patients, compared with 54% of patients where administration was outside the operating room (odds ratio, 7.74; 95% CI = 6.49 to 9.22). CONCLUSIONS: Early PA administration accounted for the majority of inappropriately timed PA. Efforts to improve performance on this measure should focus on administering antibiotics in the operating room. PMID- 17116548 TI - Rural versus urban inpatient case-mix differences in the US. AB - BACKGROUND: Preparation of surgeons for practice in rural settings is hindered by limited knowledge of case-mix differences between rural and nonrural surgical practices. Although surgical practice in isolated rural areas is believed to be very different from urban practice, little is known about actual inpatient case mix differences. STUDY DESIGN: We performed a retrospective, descriptive comparison of inpatient general surgical procedures performed at rural versus urban hospitals in the US using the Nationwide Inpatient Sample database (2000 to 2001). Rural versus urban geographic designations were based on Rural-Urban Commuting Area codes developed by the Rural Health Research Institute. Inpatient surgical procedures were aggregated by the Clinical Classifications Software based on ICD-9-CM procedure codes. RESULTS: Operations on the bowel, appendix, and gallbladder constitute 61% of general surgical inpatient procedures in rural hospitals, compared with 46% in urban hospitals. Compared with urban general surgery practices, rural practices include substantially fewer operations on the stomach and esophagus (6% versus 11%), liver and pancreas (0% versus 1%), spleen and thyroid (3% versus 10%), and bowel (17% versus 19%). General surgical procedures constitute 42% of inpatient procedures in rural hospitals versus 25% in urban hospitals. A rural general surgeon more broadly trained in selected obstetric and gynecologic operations could potentially perform 66% of all inpatient procedures in rural hospitals. Addition of simple vascular cases (eg, arteriovenous fistula, vascular access), head and neck operations, amputations, and nephrectomies could increase this potential to 71% of all cases. CONCLUSIONS: Rural and urban general surgical inpatient case-mixes differ from each other substantially. Additional competence in a few surgical areas that are not currently emphasized in general surgical training could result in an increased role for general surgeons practicing in rural areas. PMID- 17116549 TI - Arginase activity is increased by thrombin: a mechanism for endothelial dysfunction in arterial thrombosis. AB - BACKGROUND: Earlier observations implicate arterial thrombosis causing endothelial dysfunction by decreasing nitric oxide (NO) levels. NO levels are restored by regional L-arginine supplementation in animal models. The purpose of this study was to investigate the roles of thrombus components in NO generation. STUDY DESIGN: Human umbilical vein endothelial cells were harvested and cultured. The thrombus components thrombin, thrombin receptor agonist peptide (TRAP), and fibrin were added to a media of confluent human umbilical vein endothelial cells. Endothelial nitric oxide synthase (eNOS) activity was assayed by measuring conversion of L-arginine to L-citrulline. Endothelial NOS mRNA levels were quantitated using real-time polymerase chain reaction. Cellular membrane transport of L-arginine through the y+ channel was assayed with (14)C-labeled L arginine. Arginase activity was determined as the conversion of (14)C L-arginine to (14)C urea and trapped as Na(2)(14)CO(3) for scintillation counting. Arginase protein amounts were assessed using Western blotting. RESULTS: Endothelial cells exposed to thrombin for 4 hours led to increased arginase activity. Thrombin (10 U/mL) caused a 1.6-fold increase compared with that in controls (320+/-29 microM urea/min versus 194+/-10 microM urea/min, p=0.03), and thrombin (30 U/mL) increased arginase activity 2.1-fold (398+/-27 microM urea/min, p < 0.001, versus controls); thrombin at 1 U/mL and fibrin had no effect. TRAP (50 microM) had an effect similar to that of thrombin 10 U/mL (316+/-21 microM urea/min, p < 0.01, versus controls). Protein amounts of arginase corresponded with activity levels. Neither eNOS nor inducible nitric oxide synthase (iNOS) activities were affected by exposure to thrombin and TRAP for 4 hours. Similarly, quantification of eNOS, iNOS, and endothelin-1 mRNA did not change, although CL-100, a known thrombin inducible gene, was upregulated. Finally, transport of L-arginine into endothelial cells was unaffected by thrombin, TRAP, and fibrin exposure. CONCLUSIONS: Endothelial cells exposed to thrombin have increased arginase enzymatic activity, and the remainder of NO generation capability is unaffected. L-arginine supplementation or arginase blockade may counteract endothelial dysfunction in the setting of acute arterial thrombosis. PMID- 17116550 TI - Why should surgeons care about clinical research methodology? AB - BACKGROUND: A low prevalence of high-level clinical studies in the surgical literature has been reported previously. We reviewed a recent sample of surgical publications to assess the current status of clinical research. STUDY DESIGN: A 3 month sample of journal articles in Archives of Surgery, Surgery, and Annals of Surgery in 2005 was evaluated by two independent reviewers to determine the distribution of articles in established evidence classes. RESULTS: A total of 133 publications were identified in the three journals during the time periods reviewed, including 101 clinical articles and 30 basic science articles. Among the clinical papers, there were 8 class I studies (7.9%), 34 class II studies (33.7%), and more than half were class III studies (59 of 101, or 58.4%). CONCLUSIONS: The low prevalence of high-level evidence to guide surgical management of patients persists in major general surgery journals. We believe that education about proper research methodology is not only important for researchers, but is also important for practicing surgeons, and can have important health policy implications as well. PMID- 17116551 TI - Pulmonary embolism complicating bariatric surgery: detailed analysis of a single institution's 24-year experience. AB - BACKGROUND: Morbidly obese patients undergoing bariatric procedures are at risk for pulmonary embolism (PE). Because large series are required to analyze low incidence complications, factors predictive of PE have not been clearly defined. Since 1992, short-course heparin prophylaxis, beginning immediately before operation, has been used in this center. STUDY DESIGN: Prospective data on 3,861 patients undergoing bariatric procedures between 1980 and 2004 were queried. Factors analyzed included age, gender, body mass index, interval between procedure and PE, inpatient versus outpatient status, mortality, access method (open versus laparoscopic), and comorbidities. RESULTS: PE within 60 days of operation occurred in 33 patients (23 women, 10 men), for an incidence of 0.85%. No difference in incidence was noted between open (0.84%) and laparoscopic (0.88%) groups, nor did routine prophylaxis with heparin since 1992 decrease the incidence. The interval between procedure and PE was 13.2+/-2.6 (mean +/- SEM) days (open=13.0+/-3.0 days, laparoscopic 14.1+/-6.49 days, p=0.9). One-third of PEs occurred after hospital discharge. Pulmonary embolism-related mortality was 27%. A statistically greater body mass index was noted in PE patients compared with non-PE patients (57.2+/-2.4 kg.m(2) versus 49.9+/-0.2 kg/m(2), p < 0.01, Wilcoxon rank test). Multivariate logistic regression confirmed a primary role for preoperative weight as a predictor of PE; univariate analysis suggested an increased PE risk with obesity hypoventilation syndrome, anastomotic leak, and chronic venous insufficiency. CONCLUSIONS: Data demonstrated persistence of PE risk in the anticoagulation, laparoscopic-access era at a rate similar to that in the preanticoagulation, open-access era. Because one-third of PEs occur after hospital discharge, consideration should be given to continuing anticoagulants longer and to adopting a more aggressive policy of inferior vena cava filter prophylaxis, particularly in patients with high body mass index, obese hyperventilation syndrome, and venous insufficiency. PMID- 17116552 TI - Sixteen-slice CT angiography in patients with suspected blunt carotid and vertebral artery injuries. AB - BACKGROUND: We sought to determine whether 16-slice multidetector CT angiography (CTA) has sufficient negative predictive value for use as the initial imaging examination for patients with suspected blunt carotid and vertebral artery injury (BCVI) and to estimate the positive predictive value of different screening criteria in assessing BCVI. STUDY DESIGN: We conducted a retrospective study of patients who were imaged for suspected BCVI at a Level I trauma center during 2004. The policy at our center was to evaluate all patients with specific indications with CTA, and at the discretion of the clinical service, four-vessel digital subtraction angiography (DSA) if the CTA was normal. We recorded injury grade, location, and diagnostic certainty from the imaging report. The primary outcome was the proportion of patients with normal CTA who also had normal DSA. RESULTS: Of 372 patients imaged with CTA for suspected BCVI, 271 had normal studies. Eighty-two (30%) of those with normal initial CTA were further examined with DSA, which was normal or equivocal in 75 of these 82 patients (CTA negative predictive value, 92% [95% CI, 83% to 97%]). The aggregate positive predictive value of BCVI screening criteria was 19% (95% CI, 14% to 23%). Lateral element cervical vertebral fractures and skull base fractures were the most predictive criteria. CONCLUSIONS: Multidetector CTA misses relatively few injuries and adequately supplants DSA as a screening study in patients with risk factors for BCVI. Radiologists should maintain a high degree of suspicion in patients who meet screening criteria. Optimal imaging strategies should focus on the most predictive criteria. PMID- 17116553 TI - Computed tomography-based clinical diagnostic pathway for acute appendicitis: prospective validation. AB - BACKGROUND: Despite our regular use of CT for suspected appendicitis, a recent study at our institution demonstrated a negative appendectomy (NA) rate similar to our rate 15 years earlier. Based on analysis of this data, a diagnostic pathway was implemented prospectively for adult patients with possible appendicitis. STUDY DESIGN: Rates of CT, NA, and perforation for nonpregnant patients over 14 years of age undergoing appendectomy between August 2001 and August 2002 (PRE) were compared with prospectively collected pathway data (PATH, August 2004 to August 2005). All PATH patients were evaluated by a surgeon before CT. All females underwent CT. CT was obtained in male patients with low suspicion or pain for longer than 48 hours. After negative CT, patients were discharged from the emergency department or admitted for observation. RESULTS: There was a substantial decrease in NA rate after implementation of the pathway (4% PATH [8 of 183] versus 16% PRE [31 of 196], p < 0.001), without a change in the rate of perforation (11% PATH [20 of 175] versus 8% PRE [13 of 165], p = 0.28) or frequency of preoperative CT (59% PATH [108 of 183] versus 60% PRE [118 of 196], p = 0.84). CONCLUSIONS: Frequent appendiceal CT alone does not ensure surgical diagnostic accuracy. CT need not be used in all patients to achieve very low NA rates. An evidence-based diagnostic algorithm incorporating early surgical evaluation, objective criteria for preoperative CT, deterrence of early operation after negative CT, and use of CT to facilitate safe discharge led to substantial improvements in the care of our adult population presenting with possible appendicitis. PMID- 17116554 TI - Use of the round ligament of the liver to decrease pancreatic fistulas: a novel technique. AB - BACKGROUND: The reported pancreatic anastomosis fistula rate for pancreaticoduodenectomy, distal pancreatectomy, or enucleation is 2% to 27%. We hypothesized that reinforcement with a vascular pedicle would decrease the number of fistulas. We report a novel technique: the use of the round ligament of the liver to reinforce the pancreatic anastomosis after resection. STUDY DESIGN: Patients undergoing resection from January 1, 2000 until August 8, 2005, at a tertiary referral center, were followed in a retrospective cohort study. The round ligament of the liver was disconnected from the abdominal wall, from the umbilicus to the liver. After pancreatic resection, it was sutured to the anastomosis or closure. A pancreatic fistula was defined as follows: Jackson Pratt (JP) drainage>50 mL/d, after the fifth postoperative day, with amylase>3 times the serum level; reexploration for a fistula; postoperative pseudocyst; or death from sepsis with a presumed fistula. RESULTS: In 95 patients, we were able to mobilize the round ligament and use it as a vascular pedicle. The overall fistula rate for the series was 5.3% (5 of 95) and for pancreaticoduodenectomy it was 8.8% (5 of 57). There were no fistulas within the distal pancreatectomy and enucleation group (n=38). Importantly, there was no mortality from pancreatic fistula in the studied patients and no need for operative intervention for a fistula. CONCLUSIONS: We present a novel technique to prevent pancreatic fistula. Although randomized trials are necessary, it appears that the use of the round ligament as a vascular pedicle for reinforcing the pancreatic anastomoses and resections results in a very low number of pancreatic fistulas. PMID- 17116555 TI - Surgical risk factors, morbidity, and mortality in elderly patients. AB - BACKGROUND: The aging population of the United States results in increasing numbers of surgical operations on elderly patients. This study observed aging related to morbidity, mortality, and their risk factors in patients undergoing major operations. STUDY DESIGN: We reviewed our institution's American College of Surgeons National Surgical Quality Improvement Program database from February 24, 2002, through June 30, 2005, including standardized preoperative, intraoperative, and 30-day postoperative data points. This required review and analysis of the prospectively collected data. We examined patient demographics, preoperative risk factors, intraoperative risk factors, and 30-day outcomes with a focus on those aged 80 years and older. RESULTS: A total of 7,696 surgical procedures incurred a 28% morbidity rate and 2.3% mortality rate, although those older than 80 years of age had a morbidity of 51% and mortality of 7%. Hypertension and dyspnea were the most frequent risk factors in those aged 80 years and older. Preoperative transfusion, emergency operation, and weight loss best predicted morbidity for those 80 years of age and older. Operative duration predicted "other" postoperative occurrences and emergent case status predicted respiratory occurrences across all age groups. Preoperative impairment of activities of daily living, emergency operation, and increased American Society of Anesthesiology classification predicted mortality across all age groups. A 30-minute increment of operative duration increased the odds of mortality by 17% in patients older than 80 years. Postoperative morbidity and mortality increased progressively with increasing age. Age was statistically significantly associated with morbidity (wound, p = 0.021; renal, p = 0.001; cardiovascular, p = 0.0004; respiratory, p < 0.0001) and mortality (p = 0.001). CONCLUSIONS: Although several risk factors for postoperative morbidity and mortality increase with age, increasing age itself remains an important risk factor for postoperative morbidity and mortality. PMID- 17116556 TI - Analysis of prognostic factors in seventy patients having a complete cytoreduction plus perioperative intraperitoneal chemotherapy for carcinomatosis from colorectal cancer. AB - BACKGROUND: Although lymph node and liver metastases are recognized as indications for resection of metastatic disease from colorectal cancer, carcinomatosis has not traditionally been regarded as having surgical treatment options. Reports have suggested that complete surgical removal of carcinomatosis combined with thorough irrigation of the peritoneal cavity with chemotherapy could result in longterm survival in selected patients. Proper selection factors are important because palliative surgery in these patients has not proved beneficial. STUDY DESIGN: From a database of 156 patients with carcinomatosis from colorectal cancer, a retrospective analysis of data prospectively recorded in 70 patients with complete cytoreduction was performed. Eleven clinical and treatment factors were studied in univariate and multivariable analyses using survival as an end point. RESULTS: By univariate analysis, patients with peritoneal cancer index (PCI) of<20 had a median survival of 41 months compared with 16 months for patients with PCI>20 (p=0.004). The difference in negative versus positive lymph nodes was also significant; differences in survival that were improved but not significant were present for age greater than 30 years, mucinous histology, location within the colon versus rectum, and absence of an adverse factor such as cancer perforation or obstruction present at the time of primary cancer resection. Only PCI<20 versus PCI>20 and lymph node status were significant in the multivariable analysis. CONCLUSIONS: Favorable longterm results of complete cytoreduction in patients treated for carcinomatosis are associated with a limited volume of carcinomatosis observed at the time of cytoreduction and in patients with negative lymph nodes at the time of primary operation. PMID- 17116557 TI - Estimated height, weight, and body mass index: implications for research and patient safety. AB - BACKGROUND: Research suggests that weight influences patient care and outcomes. Health-care providers (HCPs) sometimes rely on patient self-reports or HCP estimates of height and weight. The purpose of this study was to determine the accuracy of self-reported height and weight and HCP estimations of height, weight, and body mass index (BMI) classification when compared with measured height, weight, and calculated BMI. STUDY DESIGN: Attending physicians, residents, and nurses provided height and weight estimates along with BMI categorizations for 110 trauma patients at a large, teaching hospital with a Level I trauma center. Patients provided reports of their heights and weights. Measured heights and weights were obtained with appropriate calibrated devices, and BMIs were calculated. Estimates and categorizations were then compared with measured and calculated values. RESULTS: HCPs were 41% and 53% accurate in estimating height and weight, respectively. Self-reports had higher accuracy (69% and 92%, respectively) but still resulted in a BMI misclassification of 32%. Twenty-two percent of patient self-reports were unobtainable. When HCPs attempted to categorize a patient into a BMI group, the accuracy was 56%. Functioning, calibrated instruments for measuring height and weight were frequently unavailable in relevant hospital locations. CONCLUSIONS: This study demonstrated that HCPs' estimates of height, weight, and BMI category are highly inaccurate. Patient self-reports are better, but are unavailable at times. Objective measurements with calibrated instruments are necessary for accuracy in research studies and for patient safety in clinical practice. Efforts to ensure the availability of calibrated instruments may be necessary in the hospital setting. PMID- 17116558 TI - A single institution review of new breast malignancies identified solely by sonography. AB - BACKGROUND: The role of sonography as a sole identifier of breast malignancy remains undetermined. Currently, the American College of Radiology Imaging Network (ACRIN) trial, protocol 6666, is accruing high risk patients, with dense breast tissue on mammography, to evaluate the performance of screening sonography. STUDY DESIGN: We conducted a retrospective single institution review to identify the detection rate of nonpalpable, mammographically occult breast malignancies identified solely by sonography. RESULTS: A total of 34,694 breast sonograms were performed at our outpatient radiology affiliate between April 1998 and April 2006. This number includes unilateral and bilateral examinations, and, 6-month followup examinations. Computerized databases and individual charts were retrospectively reviewed. Sonographic and mammographic findings were compared to determine concordance of identified lesions. Four thousand ninety-one ultrasound guided biopsies, (fine needle aspiration biopsy [FNA] and core needle biopsy [CNB]) were performed over the 8-year period, and 452 malignancies were identified. Discarded fluid contents of pure cyst aspirations were not included in this number. Seven percent (32) of the biopsy-proved malignancies were mammographically occult, of which 59% (19) were palpable, and 31% (10) were not palpable. Of all cancers diagnosed, 2.2% were identified solely by sonography. The number of cancers identified solely by sonography relative to the total number of sonograms done was 0.03%. CONCLUSIONS: Given these results, we advocate the selective use of sonography in appropriate populations, namely, in those with palpable findings, mammographic abnormalities, and in women with dense tissue who have personal or family history of breast cancer. PMID- 17116559 TI - Hemodynamic patterns of blunt and penetrating injuries. AB - BACKGROUND: The aims of this prospective observational study were to describe early hemodynamic patterns of blunt and penetrating truncal injury and to evaluate outcomes prediction using noninvasive hemodynamic monitoring with a mathematical model tested against actual in-hospital outcomes. The hypothesis was that traumatic shock is a circulatory disorder that can be monitored by noninvasive hemodynamic parameters that reflect cardiac, pulmonary, and tissue perfusion functions. STUDY DESIGN: The cardiac index (CI), heart rate (HR), mean arterial pressure (MAP), pulse oximetry (SapO(2)), transcutaneous oxygen tension indexed to FiO(2) (PtcO(2)/FiO(2)), and carbon dioxide (PtcCO(2)) tensions were monitored beginning shortly after emergency department admission in 657 emergency patients with severe blunt and penetrating chest, abdominal, and extremity trauma. Of these, 113 patients had associated head injury, and these patients also were analyzed separately. A search and display mathematical model, with a decision support program, was based on continuous online, real-time, noninvasive hemodynamic monitoring. RESULTS: There were similar patterns in the blunt and penetrating injuries; the cardiac index, mean arterial pressure, pulse oximetry, transcutaneous oxygen tension indexed to FiO(2), and survival probability values of the survivors were significantly higher (p < 0.01) than the corresponding values of those who died, although heart rate and carbon dioxide tension were higher in the nonsurvivors during the first 24 hours after their emergency department admission. These patterns occurred more rapidly in patients with penetrating injuries. After initial resuscitation in the emergency department, results were correlated with actual outcomes at hospital discharge and found to be 88% correct. CONCLUSIONS: Early noninvasive hemodynamic monitoring with a computerized information system provided a feasible pattern recognition program for outcomes prediction and therapeutic decision support. PMID- 17116560 TI - Impact of cirrhosis on outcomes in trauma. AB - BACKGROUND: Cirrhosis as an independent predictor of poor outcomes in trauma patients was identified in 1990. We hypothesized that the degree of preinjury hepatic dysfunction is, by itself, an independent predictor of mortality. STUDY DESIGN: The trauma registry at our Level I trauma center was queried for all ICD 9 codes for liver disease from 1999 to 2003, and patients were categorized as having Child-Turcotte-Pugh (CTP) class A, B, or C cirrhosis. Data analyzed included age, mechanism of injury, Abbreviated Injury Score (AIS), Injury Severity Score (ISS), Glasgow Coma Score (GCS), hospital length of stay, ventilator days, procedures performed, transfusion of blood products, admission lactate, base deficit, and mortality. Trauma Related Injury Severity Score (TRISS) methodology was used to calculate the probability of survival. Outcomes data were analyzed, and statistical comparison was performed using group t-test. RESULTS: Of the 50 patients meeting study criteria, 31 had alcohol-related cirrhosis, 18 had a history of hepatitis C, and 1 had cryptogenic cirrhosis. Twenty (40%) met CTP A classification, 16 (32%) met CTP B criteria, and 14 (28%) had CTP class C cirrhosis. One death occurred in the CTP A and B groups. Comparison between the five survivors and nine nonsurvivors from CTP class C showed no statistical significance in terms of age, ISS, TRISS, or GCS. CONCLUSIONS: The mortality rate for class C cirrhotic patients posttrauma continues to be higher than that predicted by TRISS, although patients with less severe hepatic dysfunction do not appear to have significantly lower than predicted survival. The degree of hepatic dysfunction remains an independent predictor of mortality and CTP C criteria must be considered when determining outcomes for patients posttrauma. PMID- 17116561 TI - Efficacy of continuous wound catheters delivering local anesthetic for postoperative analgesia: a quantitative and qualitative systematic review of randomized controlled trials. PMID- 17116562 TI - Thoracic damage-control operation: principles, techniques, and definitive repair. PMID- 17116563 TI - The surgeon and acts of civilian terrorism: blast injuries. PMID- 17116564 TI - The state of US trauma systems: public perceptions versus reality--implications for US response to terrorism and mass casualty events. AB - BACKGROUND: Injury has long been identified as the number one killer of Americans under the age of 34, and establishment of regional trauma systems and centers incorporating primary, secondary, and tertiary care and injury-prevention strategies has proved to be a vital element in reducing injury-related sequelae, deaths, and even costs. Despite these facts, trauma system development has not been given priority for funding in many local and state governments and only intermittently at the federal level. Consequently, many of the nation's trauma centers are strapped for funds to provide emergency care to their patients. STUDY DESIGN: In response to a 2002 Health Resources and Services Administration (HRSA) report, which identified public support as a key element in the success of trauma system development in states and communities across the United States, a Harris Interactive study was undertaken in the fall of 2004 to determine the public's attitudes, awareness, and knowledge concerning the nature and availability of trauma care and systems of trauma care. Results of the poll were contrasted with current data on the state of US trauma systems to determine the degree of correspondence. RESULTS: Results of the poll indicated that fully 61% of the American public does not know that injury is the leading cause of death for those aged 1 to 34, and most believe that a trauma system is in place in every state. Almost two-thirds of the American public is confident of receiving the best medical care in the event of serious injury and would be seriously concerned if no trauma center were nearby. But only eight states have fully developed trauma systems, and most states have no federal funding or infrastructure in place for managing the aftermath of a natural disaster or terrorist event. These and other objective data reveal the mismatch between public perceptions and reality. CONCLUSIONS: Although almost 90% of Americans believe that state trauma systems and hospitals should have a coordinated trauma response, this has not been made a national priority. Trauma systems must be adequately developed and supported to fulfill the public's expectation to receive the best possible care if seriously injured, and to ensure readiness for mass casualty and terrorist incidents. PMID- 17116565 TI - Delivery of emergency neurosurgical care. PMID- 17116566 TI - Spontaneous isolated dissection of the superior mesenteric artery. PMID- 17116567 TI - Aberrant right hepatic duct originating from the right posterior duct with low insertion into the common bile duct. PMID- 17116568 TI - First photographic surgical textbook. PMID- 17116569 TI - Screening endoscopy: cost-effective? PMID- 17116571 TI - Sustainable reduction of bioreactor contamination in an industrial fermentation pilot plant. AB - Facility experience primarily in drug-oriented fermentation equipment (producing small molecules such as secondary metabolites, bioconversions, and enzymes) and, to a lesser extent, in biologics-oriented fermentation equipment (producing large molecules such as recombinant proteins and microbial vaccines) in an industrial fermentation pilot plant over the past 15 years is described. Potential approaches for equipment design and maintenance, operational procedures, validation/verification testing, medium selection, culture purity/sterility analysis, and contamination investigation are presented, and those approaches implemented are identified. Failure data collected for pilot plant operation for nearly 15 years are presented and best practices for documentation and tracking are outlined. This analysis does not exhaustively discuss available design, operational and procedural options; rather it selectively presents what has been determined to be beneficial in an industrial pilot plant setting. Literature references have been incorporated to provide background and context where appropriate. PMID- 17116572 TI - Hydrogen bonding of water-ethanol in alcoholic beverages. AB - An alcoholic beverage is a type of water-ethanol solution with flavor and taste. The properties of the hydrogen bonding of water-ethanol in alcoholic beverages have not been clarified sufficiently. We investigated factors that could affect the hydrogen-bonding structure of water-ethanol on the basis of proton nuclear magnetic resonance (1H NMR) chemical shifts of the OH of water-ethanol and Raman OH stretching spectra. Not only acids (H+ and HA: undissociated acids) but also bases (OH- and A-: conjugate-base anions from weak acids) strengthened the hydrogen-bonding structure of water-ethanol. It was also demonstrated that the hydrogen bonding is strengthened by chemical components in alcoholic beverages (whiskey, Japanese sake, shochu). It can be suggested that hydrogen-bonding donors as well as acceptors in alcohol beverages, which exist as the initial components or are gained later on, should cause the tight association between water and ethanol molecules. PMID- 17116573 TI - Removal of p-xylene with Pseudomonas sp. NBM21 in biofilter. AB - As a p-xylene (p-Xyl)-degrading microorganism, Pseudomonas sp. NBM21 was isolated from an activated sludge of a wastewater treatment plant. NBM21 degraded p-Xyl, m xylene, benzene and toluene, but not o-xylene, ethylbenzene (Eb) and styrene. NBM21 was inoculated to a biofilter with Biosol as a packing material and p-Xyl removal was operated for 105 d under sterile and nonsterile conditions. The maximum elimination capacities for p-Xyl at higher than 90% removal efficiency were 160 g/m3/h and 150 g/m3/h under nonsterile and sterile conditions, respectively. A high load of Eb adversely affected to the removal of xylene. PMID- 17116574 TI - A knockout strain of CPR1 induced during fermentation of Saccharomyces cerevisiae KNU5377 is susceptible to various types of stress. AB - To investigate the tolerance factor of Saccharomyces cerevisiae KNU5377 against various types of environmental stress during fermentation, we identified the protein that is upregulated at high temperatures. The highly upregulated protein was high-score-matched as a cytoplasmic peptidyl-prolyl cis-trans isomerase, cyclophilin (Cpr1p), by matrix-assisted laser desorption ionization time-of flight mass spectrometry (MALDI-TOF). We constructed a CPR1-deleted KNU5377 strain (KNU5377Y cpr1Delta) to determine the roles of the protein under fermentative or stress condition. The growth of the S. cerevisiae KNU5377Y cpr1Delta strain was completely inhibited under the following conditions: heat (40 degrees C), hydrogen peroxide (20-30 mM), menadione (0.3 mM), ethanol (16%), sulfuric acid (5 mm), and lactic acid (0.4-0.8%). However, the wild-type and cpr1Delta mutant of S. cerevisiae BY4741 as a positive control did not show differences in sensitivity to stress. It is interesting to note that the wild type KNU5377Y and KNU5377Y cpr1Delta mutant showed high sensitivity against various stresses, particularly, acid stress such as in the presence of sulfuric and lactic acid. Although the alcohol fermentation rate of the KNU5377Y cpr1Delta mutant markedly decreased with an increase in temperature up to 40 degrees C, we observed no decrease in that of the wild-type strain under the same conditions. These results suggest that CPR1 contributes to the stress tolerance of KNU5377 against various types of environmental stress caused during fermentation, thus leading to the physiological role of maintaining an alcohol fermentation yield, even at high temperatures such as 40 degrees C. PMID- 17116575 TI - Production of scFv-Fc fusion protein using genetically manipulated quails. AB - The use of transgenic avian species as a transgenic bioreactor for the production of recombinant proteins has been proposed. In recent years, although various procedures for generating transgenic chickens have been reported, the expression of a useful protein at a commercially feasible level has rarely been attained. In this study, we injected a concentrated retroviral vector into quail embryos to generate genetically manipulated quails that produce recombinant proteins. We found that transgene expression in the whole body at a high level was observed for viral injection into the heart of the developing embryos after a 48-h incubation. For the practical production of a useful protein, a retroviral vector encoding an anti-prion scFv-Fc gene under the control of the beta-actin promoter was injected into quail embryos. The quails that hatched stably produced scFv-Fc at a high level in their serum and egg white. The production of scFv-Fc was maintained throughout the breeding period. scFv-Fc purified from the egg white retained the antigen-binding activity. This system exhibited the potential of transgenic quails for the commercial production of recombinant proteins. PMID- 17116576 TI - Culture of mouse embryonic stem cells on photoimmobilized polymers. AB - Mouse embryonic stem (ES) cells were cultured on four types of polymer with different surface properties. The polymers were poly(acrylic acid), polyallylamine, gelatin, and poly(2-methacryloyloxyethyl phosphorylcholine-co methacrylic acid) (PMAc50), and were coupled with azidophenyl groups and photoimmobilized on conventional polystyrene cell-culture dishes. Mouse ES cells were cultured on the immobilized polymer surfaces, and cell morphology, cell growth, staining for alkaline phosphatase, activation of the transcription factor stat3, and expression of the octamer-binding protein 3/4 (Oct3/4) transcription factor and the zinc finger-containing transcription factor (GATA4) were observed. Morphology and growth rate were significantly affected by the polymer surface properties. The ES cells attached to gelatin or polyallylamine surfaces; however, colonies formed on the former but not the latter. In addition, significant enhancement of growth was observed on the gelatin surface. In contrast, ES cells aggregated to form an embryoid body on the photoimmobilized poly(acrylic acid) surface and the PMAc50 surface, although cell growth was reduced. Significant enhancement of aggregation of ES cells on the PMAc50 surface was observed in morphology and gene expression analyses. PMID- 17116577 TI - Purification and properties of glutamine synthetase from Hydrogenobacter thermophilus TK-6. AB - Hydrogenobacter thermophilus TK-6, a thermophilic and obligately chemoautotrophic bacterium, assimilates ammonium using glutamine synthetase (GS). GS was purified using three chromatography steps. The purified GS was found to belong to GS type I on the basis of its subunit composition and molecular weight. The Mg2+ dependent activity of this GS significantly increased after incubation with phosphodiesterase, indicating that GS is subject to adenylyl/deadenylyl regulation, a posttranslational modification system reported mainly among enterobacteria. The degree of this posttranslational modification changed depending on growth phase, confirming that adenylyl/deadenylyl regulation functions in vivo. Interestingly, the Km for glutamate of H. thermophilus GS was significantly higher than those of other organisms, suggesting that GS activity is affected by intracellular glutamate concentration. PMID- 17116578 TI - Effects of medium composition on production of 5-aminolevulinic acid by recombinant Escherichia coli. AB - The recombinant Escherichia coli BL21(DE3) harboring hemA from Agrobacterium radiobacter, which was engineered in our previous work, was used for the extracellular production of 5-aminolevulinic acid (ALA). The effects of various physiological factors, such as the concentrations of precursors (glycine, succinic acid and glucose) and the inhibitor 5-aminolevulinate dehydratase (levulinic acid), on the ALA accumulation in the fermentation broth were investigated in both shake flasks and a jar fermentor. Among these precursors, glycine exhibited the strongest ability to inhibit cell growth, while glucose mainly inhibited ALA formation. The optimum initial concentrations of glycine, succinic acid and glucose were found to be 2.0, 10.0 and 2.0 g/l, respectively. Levulinic acid (LA; 30 mM) was fed to the fermentation broth at the end of the exponential cell growth phase (about 8 h), and the intracellular activity of ALA dehydratase was efficaciously suppressed. Repeating the optimum composition of the medium in a stirred tank fermenter resulted in 1.49 g/l ALA. Furthermore, the fed batch of the precursors and inhibitor further increased ALA production up to 3.01 g/l. PMID- 17116579 TI - Utilization of Shochu distillery wastewater for production of polyunsaturated fatty acids and xanthophylls using thraustochytrid. AB - The industrial production of barley shochu, a distilled alcoholic beverage, results in distillery waste that is currently incinerated or disposed of in landfills, causing environmental pollution. The supernatant of distillery waste contains organic matter such as proteins ( approximately 2.5%) and amino acids ( approximately 0.2%). This study demonstrates that the utilization of distillery wastewater as a sole nitrogen source enables a marine thraustochytrid, Schizochytrium sp. strain KH105, to propagate and accumulate valuable lipids including docosahexaenoic acid (DHA) and astaxanthin. Under optimized culture conditions, the highest DHA and astaxanthin yields were obtained at 3.4 g/l and 7.7 mg/l, respectively, after 4 or 5 d of cultivation in a 3-l jar fermentor. The chemical oxygen demand of the wastewater was reduced by 35%. About 67% of crude protein content and 85% of total free amino acid content also decreased in the culture supernatant. The thraustochytrid therefore serves to upgrade the distillery by-product to one containing value-added lipids for functional foods as well as to regulate the environmental contamination. PMID- 17116580 TI - Effects of temperature and hydraulic retention time on anaerobic digestion of food waste. AB - A modified three-stage methane fermentation system was developed to digest food waste efficiently. This system consisted of three stages: semianaerobic hydrolysis, anaerobic acidogenesis and strictly anaerobic methanogenesis. In this study, we examined the effects of temperature and hydraulic retention time (HRT) on the methanogenesis. Operation temperature was adjusted from 30 degrees C to 55 degrees C, and the HRTs ranged from 8 to 12 d. The rate of soluble chemical oxygen demand (sCOD) removal correlated with digestion time according to the first-order kinetic model developed by Grau et al. [Water Res., 9, 637-642 (1975)]. With liquor food waste, thermophilic digesters showed a higher rate of sCOD removal than mesophilic digesters. The rates of biogas and methane production by thermophilic digesters were higher than those by mesophilic digesters regardless of HRT. Although maximum biogas production occurred when an HRT of 10 d was used, the methane yield was the highest in the reactor when an HRT of 12 d was used (223 l CH4/kg sCODdegraded). However, digestion stability decreased when an HRT of 8 d was used. The concentration of NH3-N generated in this experiment did not inhibit anaerobic digestion. PMID- 17116581 TI - Enhancement of recombinant soluble dengue virus 2 envelope domain III protein production in Escherichia coli trxB and gor double mutant. AB - The dengue virus is currently the most important flavivirus causing human diseases in the tropical and subtropical regions of the world. The envelope protein domain III of dengue virus type 2 (D2EIII), which induces protective and neutralizing antibodies, was expressed as an N-terminal fusion to a hexa histidine tag in Escherichia coli. The expression of recombinant D2EIII of 103 amino acids in the soluble form can be achieved using suitable host strains, such as Origami, at a low induction temperature of 18 degrees C. The enhanced production of the soluble protein could be attributed to the thioredoxin reductase (trxB) and glutathione reductase (gor) double mutations in the Origami genome. The soluble and refolded D2EIII proteins were recognized by different antibodies including human patient antiserum. The immunization of rats with soluble D2EIII protein elicited the production of antibodies that could recognize the D2EIII protein in the D2EIII precursor protein and in C-terminal truncated dengue envelope protein type 1-4. Thus, this protein production system is suitable for the production of authentic recombinant dengue proteins that may be used in the diagnosis of the dengue virus infection or in vaccine development. PMID- 17116582 TI - Rice protein digestion by sake koji enzymes: comparison between steamed rice grains and isolated protein bodies from rice endosperm. AB - The digestion of proteins in steamed rice grains by sake koji enzymes under simulated sake mash conditions was analyzed by comparing the hydrolysis of steamed rice grains and heat-treated protein bodies (PBs) isolated from seven rice samples including four endosperm-storage protein mutants. The disappearance of peptides in the digest of isolated PBs was faster than that of steamed rice grains; however, more insoluble proteins formed in the case of isolated PBs. Not all of the hydrolyzed PB proteins were immediately solubilized in the digestion tests. High-molecular-weight peptides were more abundant in the solubilized digest of steamed rice grains than in that of isolated PBs. Variance in Ile, Ser, Glu, and Gly levels in the digest of steamed rice grains was relatively high among the seven samples, but was not found to be high in digests of isolated PBs. These results indicate that factors that may be derived from the steamed rice grains profoundly affect the digestion of proteins in steamed rice grains by sake koji enzymes. PMID- 17116583 TI - Nitrogen removal from wastewater using simultaneous nitrate reduction and anaerobic ammonium oxidation in single reactor. AB - The effects of C/N ratio and total organic carbon (TOC) loading on nitrogen removal through simultaneous nitrate reduction and anaerobic ammonium oxidation in a single reactor were examined. Granular sludge taken from a methane fermentation reactor was placed in an upflow reactor and supplied with synthetic wastewater containing nitrate at a C/N ratio of 1 to grow heterotrophic denitrifying bacteria. When nitrogen removal ratio reached 30%, anammox sludge attached to nonwoven-carrier was added into the same reactor and then ammonia was added to the synthetic wastewater. Nitrogen removal ratio was markedly increased to 80-94%. In this system, nitrogen removal ratio was affected by C/N ratio and TOC loading, not by the amount of granular sludge. A stable isotopic analysis using 15N-labeled nitrate showed that N2 gas was formed by anammox reaction. PMID- 17116584 TI - Characterization of alkylphenol degradation gene cluster in Pseudomonas putida MT4 and evidence of oxidation of alkylphenols and alkylcatechols with medium length alkyl chain. AB - Alkylphenols (APs) are ubiquitous contaminants in aquatic environments and have endocrine disrupting and toxic effects on aquatic organisms. To investigate biodegradation mechanisms of APs, an AP degradation gene cluster was cloned from a butylphenol (BP)-degrading bacterium, Pseudomonas putida MT4. The gene cluster consisted of 13 genes named bupBA1A2A3A4A5A6CEHIFG. From the nucleotide sequences, bupA1A2A3A4A5A6 were predicted to encode a multicomponent phenol hydroxylase (PH), whereas bupBCEHIFG were expected to encode meta-cleavage pathway enzymes. A partial sequence of a putative NtrC-type regulatory gene, bupR, was also found upstream of the gene bupB. This result indicates that APs can be initially oxidized into alkylcatechols (ACs), followed by the meta cleavage of the aromatic rings. To confirm this pathway, AP degradation tests were carried out using the recombinant P. putida KT2440 harboring the PH genes (bupA1A2A3A4A5A6). The recombinant strain oxidized 4-n-APs with an alkyl chain of up to C7 (< or = C7) efficiently and also several BPs including those with an alkyl chain with some degree of branching. Therefore, it was found that PH had a broad substrate specificity for APs with a medium-length alkyl chain (C3-C7). Moreover, the cell extract of a recombinant Escherichia coli harboring bupB (a catechol 2,3-dioxygenase gene) converted 4-n-ACs with an alkyl chain of < or = C9 into yellow meta-cleavage products with a maximum absorbance at 379 nm, indicating that the second step enzyme in this pathway is also responsible for the degradation of ACs with a medium-length alkyl chain. These results suggest that MT4 is a very useful strain in the biodegradation of a wide range of APs with a medium-length alkyl chain, which known nonylphenol-degrading Sphingomonas strains have never degraded. PMID- 17116585 TI - Analysis of essential amino acid residues for catalytic activity of glutaminase from Micrococcus luteus K-3. AB - Structural-based mutational analysis of salt-tolerant glutaminase from Micrococcus luteus K-3 (Micrococcus glutaminase) revealed that three amino acid residues, S64, K67, and E160, were essential to a catalytic reaction. The result suggested that Micrococcus glutaminase had a possible catalytic mechanism similar to class A beta-lactamase rather than glutaminase-asparaginase from Pseudomonas 7A. PMID- 17116586 TI - Entomogenous fungi that produce 2,6-pyridine dicarboxylic acid (dipicolinic acid). AB - An inhibitor of the prophenoloxidase activation using extract from a silkworm pupa was isolated from a culture filtrate of Cordyceps militaris and identified as dipicolinic acid (DPA). The production of DPA in Clavicipitaceae fungi was examined. Entomogenous fungi that produce DPA were integrated into one group by a phylogenetic analysis based on 18S rDNA. It is suggested that the group acquired an ability to produce DPA during its evolution from plant pathogenic fungi to entomogenous fungi. PMID- 17116587 TI - Enhancement of water-solubility and bioactivity of paclitaxel using modified cyclodextrins. AB - We investigated the aqueous solubility of paclitaxel using 11 kinds of cyclodextrins (CDs) and the bioactivity of the paclitaxel-CD inclusion complex. 2,6-Dimethyl beta-cyclodextrin was the most effective and its solubility was 2.3 mM in a 0.1 M 2,6-dimethyl beta-cyclodextrin aqueous solution. The inclusion complex of paclitaxel and 2,6-dimethyl beta-cyclodextrin had a 1.23-fold polymerization activity as paclitaxel in a tubulin assay. PMID- 17116588 TI - Application of femtosecond laser ablation for detaching grown protein crystals from glass capillary tube. AB - We developed a novel technique for detaching protein crystals from glass capillary tube using the counter diffusion crystallization technique by femtosecond laser irradiation. X-ray diffraction analysis demonstrated that femtosecond laser irradiation has little effect on crystallinity. This technique will contribute to progress in structural genomics as a powerful tool. PMID- 17116589 TI - Consumer research for development of educational messages for the MyPyramid Food Guidance System. AB - OBJECTIVE: To assess consumer understanding and use of messages from the original Food Guide Pyramid and potential concepts for a revised Food Guidance System. DESIGN: Focus groups conducted in two phases, in 2002 and 2004. SETTING: Market research facilities in Baltimore, Chicago, and Houston. PARTICIPANTS: Phase I, 178 participants in 18 groups: 6 of general adult consumers, 4 of adults over 60 years of age, 4 of food stamp recipients, and 4 of overweight adults. Phase II, 75 participants in 8 groups: 4 of younger adults and 4 of older adults. PHENOMENON OF INTEREST: Understanding and use of original Pyramid symbol and messages and potential concepts for a revised food guidance system. ANALYSIS: Focus group sessions were audiotaped and transcribed. Content analysis summarized comments into meaningful themes. RESULTS: Key concepts of the original Pyramid were widely understood, but specific knowledge was limited and misunderstandings common, especially related to servings and food group placement. Detailed information about whole grains, types of fats, vegetable subgroups, and physical activity was lacking. CONCLUSIONS AND IMPLICATIONS: While consumers are aware of general concepts about healthy eating, they lack specific knowledge to help them implement recommendations. Educators can help by providing consumers with concrete examples and specific information. PMID- 17116590 TI - Developing the consumer interface for the MyPyramid Food Guidance System. AB - OBJECTIVE: To assess consumer response to potential graphics, slogans, and messages for the consumer interface of the MyPyramid Food Guidance System. DESIGN: Qualitative research conducted in two phases, composed of focus groups and Web-TV testing. SETTING: Professional market research facilities in Baltimore, MD, and Chicago, IL, and Web-TV. PARTICIPANTS: Phase 1,77 adults in 10 groups; Phase 2,407 adults via Web-TV. PHENOMENON OF INTEREST: Consumer response to potential graphic images, slogans, and messages for the Food Guidance System. ANALYSIS: A content analysis was used to summarize comments from focus groups and Web-TV tests into meaningful themes. Frequencies were calculated for responses. RESULTS: Respondents preferred the familiarity of the pyramid shape and found graphics and slogans that were personal, active, and positive to be appealing. CONCLUSIONS AND IMPLICATIONS: The consumer interface for the MyPyramid Food Guidance System was shaped by consumer feedback that identified appealing and useful elements and avoided elements that were potentially confusing or less meaningful. Consumers preferred images and messages that were perceived as new, personal, and active, but they desired some continuity with the original Pyramid shape. PMID- 17116591 TI - Impact of using updated food consumption and composition data on selected MyPyramid Food Group nutrient profiles. AB - OBJECTIVE: To examine the changes observed in 5 nutrients of selected USDA food subgroups by partitioning the overall changes into those caused by consumption changes over time, and those caused by nutrient database revisions. DESIGN: Population-weighted estimates of food group intakes (composites) were developed using 24-hour recall data from CSFII 1994-96 and NHANES 1999-2000. Nutrient profiles of these composites were developed using Standard Reference (SR) data (SR11 and SR16-1). SUBJECTS: A total of 14,262 and 8070 individuals over the age of 2 years from CSFII and NHANES, respectively, composed the study sample. OUTCOME MEASURES: Absolute and percent change in food group nutrient content caused by food consumption changes and nutrient database updates. ANALYSIS: Changes due to consumption differences were determined by comparing nutrient profiles created with CSFII and NHANES using SR11. Changes due to nutrient database differences were determined by comparing nutrient profiles created from NHANES data using SR11 and SR16-1 nutrient values. RESULTS: Consumption differences resulted in some variations in the food group nutrient content, but a majority of the changes were associated with use of the updated nutrient database. For example, vitamin A level in the orange vegetable subgroup was increased by 2.4% owing to consumption (from CSFII to NHANES), whereas the level was decreased by 38% due to nutrient updates (from SR11 to SR16-1). CONCLUSION AND IMPLICATIONS: Consideration of the changes in nutrient databases, as well as in food consumption, is essential in monitoring both the trends in the food choices Americans make and the adequacy of their diets. PMID- 17116592 TI - MyPyramid food intake pattern modeling for the Dietary Guidelines Advisory Committee. AB - Modeling analyses using the MyPyramid intake patterns were conducted in collaboration with the 2005 Dietary Guidelines Advisory Committee in response to their research questions and to determine likely effects of possible recommendations on overall dietary adequacy. Scenarios modeled included the feasibility of using the food patterns for lacto-ovo-vegetarian diets, of varying fat levels within the patterns, and of increasing dietary flexibility through food group substitutions. Food pattern modeling was a useful tool to identify possible impacts on diet quality of potential Dietary Guidelines recommendations. Modeling analyses can help researchers explore the overall effect of specific dietary recommendations on intake patterns. PMID- 17116593 TI - MyPyramid.gov provides consumers with practical nutrition information at their fingertips. PMID- 17116594 TI - MyPyramid Tracker assesses food consumption, physical activity, and energy balance status interactively. PMID- 17116595 TI - Designing MyPyramid for Kids materials to help children eat right, exercise, have fun. PMID- 17116598 TI - Development of food intake patterns for the MyPyramid Food Guidance System. AB - OBJECTIVE: The purpose of this research was to design food intake patterns based on typical American food selections that would meet Dietary Guidelines and Dietary Reference Intake recommendations. DESIGN: Analytic process to identify appropriate amounts from each food group that together will meet nutritional goals for various age/gender groups. VARIABLES MEASURED: Projected intake of energy, 9 vitamins, 8 minerals, 8 macronutrients, and dietary fiber in each food intake pattern. ANALYSIS: Iterative comparison of nutrients in each food intake pattern to Dietary Reference Intakes and Dietary Guidelines recommendations set as goals for that pattern. RESULTS: Food intake patterns were established that met almost all nutrient goals within estimated energy needs. Intakes of vitamin E at all energy levels, potassium at lower energy levels, and sodium at higher energy levels did not meet goals. CONCLUSIONS AND IMPLICATIONS: The food intake patterns provide a foundation of food choices that will meet nutritional recommendations. They form the scientific basis for the MyPyramid Food Guidance System and can also be used as a starting point for developing other educational programs or materials. PMID- 17116599 TI - Development of food group composites and nutrient profiles for the MyPyramid Food Guidance System. AB - OBJECTIVE: To identify food selections in each MyPyramid food group or subgroup reflective of typical consumption patterns by Americans, and the nutrient intake that can be expected from consuming a specified amount of these foods from each group, in a low-fat and no-added-sugars form. DESIGN: An analytical process to identify food consumption choices within each food group and subgroup using national food consumption surveys, and to identify the expected nutrient content of each group using food composition databases. VARIABLES MEASURED: Relative consumption of foods within each food group; nutrient content for each food group and subgroup (energy plus 27 nutrients). ANALYSIS: Disaggregated foods from consumption surveys into component ingredients. Combined similar ingredients into "item clusters" and determined relative consumption of each. Calculated a consumption-weighted nutrient profile for each food group. RESULTS: Consumption weighted food intake selections and nutrient profiles were developed for all MyPyramid food groups and subgroups. CONCLUSIONS AND IMPLICATIONS: This analytical process derived food group and subgroup composites which estimate typical food choices within each MyPyramid food group. These were used to assess the adequacy of the MyPyramid food intake patterns as they were being iteratively developed. PMID- 17116600 TI - The changing contours of pediatric practice. PMID- 17116602 TI - Is there a relationship between overweight and obesity and mental health problems in 4- to 5-year-old Australian children? AB - OBJECTIVE: To investigate the relationship between overweight and obesity, and mental health problems in Australian 4- to 5-year-old children. METHODS: The study used data from wave 1 (2004) of the Longitudinal Study of Australian Children (LSAC). The participants were 4983 4- to 5-year-old children (2537 boys and 2446 girls) with a mean age of 56.9 months (standard deviation 2.6 months; range 51-67 months). Children were classified as nonoverweight, overweight, and obese on the basis of International Obesity Task Force definitions. Mental health problems were assessed by the Strengths and Difficulties Questionnaire (SDQ) completed by parents and teachers. RESULTS: Although obese 4- to 5-year-old boys had more mental health problems than nonoverweight boys, differences between the groups were small and substantially reduced when analyses controlled for children's sociodemographic characteristics. Parents reported that overweight/obese girls had more peer problems, whereas teachers reported they had more conduct problems. Children in all weight groups had mean scores within the normal range of scores on all the SDQ subscales. CONCLUSIONS: Differences in rates of mental health problems experienced by young children of different weight status appear relatively small. Higher rates of mental health problems experienced by more obese boys may reflect differences in their sociodemographic characteristics rather than their weight status per se. Policies that reduce the number of young children living in poverty or experiencing other adverse social circumstances have the potential to reduce rates of mental health problems experienced by older children with overweight/obesity. PMID- 17116603 TI - Disparities in complementary and alternative medical therapy recommendations for children in two different socioeconomic communities. AB - OBJECTIVE: To compare the content, quality, and cost of recommendations for children made by complementary and alternative medicine (CAM) retailers within 2 New York City neighborhoods of divergent socioeconomic status (SES). METHODS: Posing as consumers, researchers sought recommendations from CAM retailers for 2 clinical scenarios: 1) a febrile 6-week-old and 2) a 4-year-old with an upper respiratory infection (URI). All retailers selling CAM therapies outside the direction of a licensed provider within East Harlem (EH) and the Upper East Side (UES) were eligible and mapped. The febrile infant scenario was posed at sites in business in March (n = 23) and the URI scenario at sites that remained in business in April (n = 20) of 2004. RESULTS: In response to the febrile infant scenario, 33% of UES retailers referred to a MD, 0% to the emergency department, and 47% made other recommendations-of which 43% were not indicated. In EH, 50% referred to a MD, 5% to the emergency department, and 37% made other recommendations. The mean price of UES recommendations was Dollars 9.66, whereas EH was Dollars 2.33 (P = .04). In response to the URI scenario, 93% of UES and 83% of EH retailers made recommendations. The mean price of UES recommendations was Dollars 10.55 while EH was Dollars 4.26 (P = .002). CONCLUSIONS: Complementary and alternative medicine retailers made numerous recommendations for children, including some that were contraindicated for age. East Harlem retailers tended to refer an infant with a potentially serious condition to the emergency department or to an MD and made less expensive recommendations than their UES counterparts. PMID- 17116604 TI - Less anticipatory guidance is associated with more subsequent injury visits among infants. AB - OBJECTIVE: To describe clinician delivery of injury prevention anticipatory guidance and injury visits in a birth cohort, and to describe the association of injury prevention anticipatory guidance with subsequent injury visits. METHODS: We performed a prospective cohort study of 2610 infants born from July 1, 1998 to June 30, 1999, at an urban safety-net hospital and seen subsequently for well child care (WCC, visits = 10558) and/or injury by 16 months of age. Injury guidance was defined as the proportion of recommended injury prevention anticipatory guidance items delivered to those expected, given the WCC visits the child attended. The outcome was a first injury visit to a clinic, emergency department, or hospital. RESULTS: The injury prevention items most discussed were car seats (84%-95% of all WCC visits) and rolling over at the 2-month WCC visit (80%). Other items were addressed at 36%-69% of visits. A total of 1931 (74%) of children received > or = 50% expected injury guidance. A total of 277 children (11%) had an injury visit, primarily for minor injuries. In unadjusted analysis, children receiving < 25% expected injury guidance were more likely to have a subsequent injury visit (unadjusted odds ratio 6.2; 95% confidence interval [95% CI] 3.2-9.7). In adjusted analysis, children who received < 25% and 25%-49% expected injury guidance were more likely to have a subsequent injury visits (adjusted odds ratio [AOR] 6.6; 95% CI 3.8-11.2; and AOR 2.9, 95% CI 2.0-4.3, respectively). CONCLUSIONS: Disadvantaged children whose families received less injury guidance than other children in their cohort were more likely to have a subsequent injury visit. Further studies are needed to determine whether increased injury prevention counseling reduces injury visits. PMID- 17116605 TI - Community pediatrics: gender differences in perspectives of residents. AB - OBJECTIVE: The Future of Pediatric Education II Report affirmed the importance of providing resident education in community settings. Yet we know little about related experiences of trainees and whether experiences and perspectives regarding community involvement vary by gender. We assessed gender differences in pediatric residents' involvement in and perspectives regarding community activities. METHODS: A national survey of US pediatric residents assessed residents' involvement in 14 activities before medical school and the intensity and perceived importance of involvement in 17 activities during medical school and residency. Expected future involvement 10 years hence was assessed for 11 community settings. chi2 and analysis of variance were used to examine bivariate relations by gender. Multivariate linear regression was used to model the relationship between gender and expected future involvement. RESULTS: Of the 700 respondents, 68% were women. Relative to men, more women reported exposure to child health advocacy and other community activities before and during medical school. Women and men reported similar involvement in residency, although women placed greater importance on inclusion of 16 of 17 community activities in their training. Female residents were more likely to report that current training in the community would influence their future career activities. Women anticipated greater future involvement in 6 of 11 settings. In adjusted analyses, gender remained associated with future involvement in 5 settings. CONCLUSIONS: As women come to comprise an increasing proportion of the pediatric workforce, further efforts are needed to understand the impact of gender on future involvement in community child health activities. PMID- 17116606 TI - Ambulatory Pediatric Association policy on tobacco. PMID- 17116607 TI - Pediatric residents' perceptions of community involvement prior to residency. AB - OBJECTIVES: To describe baseline perceptions of first-year pediatric residents of participating in community activities, to determine whether demographic factors are related to perceived benefits and constraints, and to identify factors associated with expected community involvement. METHODS: Pediatric residents beginning their training in the fall of 2000 to 2003 participated in a 12-item self-administered written survey as part of the national evaluation of the Dyson Community Pediatrics Training Initiative. RESULTS: Of the 612 first-year residents surveyed (90% response rate), most reported they receive personal satisfaction (92%) and gain valuable skills and knowledge (83%) from their involvement in community activities. Less than a quarter felt peer support and professional recognition were benefits. Almost two thirds reported logistics and lost personal time as constraints to community involvement. Compared with their colleagues, older residents (> 29 years) and underrepresented minority residents reported fewer constraints. Most residents (72%) expect moderate to substantial involvement in community activities after graduating. Those expecting greater involvement were more likely to report personal satisfaction, gaining valuable skills and knowledge, peer support, and the opportunity to spend time with like minded peers as benefits. CONCLUSIONS: Pediatric residents beginning their postgraduate training perceive numerous benefits from their participation in community activities and most expect a moderate degree of future community involvement. Residency directors should: 1) consider their trainees' insights from prior community involvement and 2) integrate meaningful community experiences in ways that confront logistic barriers and time constraints. PMID- 17116608 TI - Planning for health promotion in low-income preschool child care settings: focus groups of parents and child care providers. AB - OBJECTIVE: To identify potentially successful strategies, barriers, and facilitators for health promotion in preschool child care settings. METHODS: We conducted 6 focus groups including each of the following: parents of children attending child care centers and home-based family child care (2 in English, 1 in Spanish) and directors of child care centers and family child care providers (2 in English, 1 in Spanish). Systematic thematic analysis was conducted to generate themes to address study questions. RESULTS: A total of 24 parents and 45 child care providers, serving predominantly urban, low-income children in Boston, participated. Parents and child care providers agreed that in-person group discussions would be the most effective strategy for providing health education information to parents. Several barriers that could affect implementation emerged. First, some providers expressed frustration toward parents' attitudes about child safety and health. Second, there was diversity of opinion among providers on whether conducting health promotion activities was consistent with their training and role. In addition, literacy, language, and cultural barriers were identified as potential barriers to health promotion in child care. CONCLUSIONS: In order to be successful, health promotion strategies in child care settings will need to overcome tensions between providers and parents, allow professional growth of child care providers to serve in a health promotion role, and better integrate external health resources and personnel. Group sessions and peer learning opportunities that are culturally and linguistically sensitive are potentially successful strategies for implementation of health promotion interventions for many parents. PMID- 17116609 TI - Parent and teacher mental health ratings of children using primary-care services: interrater agreement and implications for mental health screening. AB - OBJECTIVE: To examine interrater agreement when screening for child mental health problems during primary-care visits. METHODS: Children aged 5 to 10 (n = 227) and one of their parents were systematically recruited from the waiting rooms of 15 primary-care sites located in Baltimore, Md; Washington, DC; and rural New York from 2002 to 2005. The parent and teacher of the child completed the Strengths and Difficulties Questionnaire to measure the child's emotions, behaviors, and functional impairment. RESULTS: Parents and teachers identified a similar proportion of children as having high symptoms (25% vs 23%) and high impairment (27% vs 32%) but rarely agreed in their assessments of specific children. Parent ratings alone missed 52% of children rated by teachers as having both high symptoms and high impairment (kappa = 0.15). Only 6% of these discrepant visits were for mental health problems, making it unlikely that teacher reports would have been solicited. CONCLUSIONS: Parent reports failed to detect half of school aged children considered to be seriously disturbed by their teachers. Efforts to improve detection of mental health problems by using screening tools in primary care may require algorithms that help providers judge when to solicit teacher reports and how to interpret conflicting information from parents and teachers. PMID- 17116610 TI - What do pediatric primary care providers think are important research questions? A perspective from PROS providers. AB - OBJECTIVE: To describe what pediatric primary care providers involved in the Pediatric Research in Office Settings (PROS) research network think are important yet inadequately addressed questions in pediatric primary care research. METHODS: A total of 1785 pediatric primary care providers in the PROS network were asked what they thought were important yet inadequately addressed areas of primary care research. We used a single, open-ended question in a mail survey. Written answers to this question were analyzed by qualitative methods to determine the main themes of interest to pediatric primary care providers. RESULTS: Overall survey response rate was 48.7%; the open-ended question yielded 1109 individual answers. Six lines of inquiry were identified as being important to these providers: (1) effective counseling techniques to use in anticipatory guidance; (2) strategies to prevent and treat obesity; (3) the effectiveness of well-child care; (4) ongoing management of patients with attention-deficit/hyperactivity disorder; (5) the role of the primary care provider in caring for children with mental health needs; and (6) optimal organization of office practices. CONCLUSIONS: The translation of research into practice may be improved by a better understanding of the needs and interests of those who see pediatric patients in the primary care setting. PMID- 17116611 TI - Revisiting of preoperative blood ordering policy--a single institute's experience in Taiwan. AB - BACKGROUND: If unnecessary blood orders can be reasonably waived, it will reduce both workload and financial expenditure. A review of the surgical blood ordering practice is, therefore, mandatory. METHODS: Routine preoperative blood orders were retrospectively audited. After receiving the requests, we usually performed only type and screen tests without cross matching until an actual need for transfusion occurred. Transfusion probability (number of patients transfused / number of procedures x 100) was calculated. One unit of donation was defined as 500 mL whole blood. If surgical procedures were associated with insignificant blood loss (number of units transfused < or = 1) and transfusion probability was less than 5%, then it was considered to be safe to disregard a preoperative blood order. RESULTS: The blood ordering practices for 5,472 patients who received various surgical procedures were reviewed over a period of 48 operation days. Neither preoperative requests for preparation of red cells nor transfusion was made in 3,482 patients. Preoperative requests for preparation of red cells were made in 1,990 patients, but only 751 (37.74%) actually received blood transfusion on the day of the operation. Analysis showed that it would have been safe to disregard a preoperative blood order for ophthalmic surgery, ear surgery, nose surgery (endoscopic sinus surgery, submucosal turbinectomy), microlaryngoscopic surgery, tracheostomy, thyroidectomy, mastectomy, laparoscopic cholecystectomy, hemicolectomy, hernioplasty, arthroscopic surgery, laminectomy, laparoscopically assisted vaginal hysterectomy, vasectomy and varicose vein surgery. CONCLUSION: A review of preoperative blood orders has identified certain surgical procedures with insignificant blood loss and low transfusion probability, for which preoperative blood orders may be safely disregarded in order to reduce unnecessary laboratory workload while not jeopardizing patient safety. PMID- 17116612 TI - Association between serum uric acid level and components of the metabolic syndrome. AB - BACKGROUND: Serum uric acid (UA) level has been suggested to be associated with factors that contribute to the metabolic syndrome. However, the association between metabolic syndrome and UA has not been elucidated. We sought to determine the association between serum UA level and the number of components that contribute to the metabolic syndrome, and which component was associated most with higher serum UA level. METHODS: A consecutive sample was taken of the health examinations of all hospital staff who were assessed between January 2004 and December 2004 in a medical center. A total of 3,065 subjects aged 18 to 81 years (635 males, 2,430 females) participated. Blood tests and all physical variables were examined using standard methods. Subjects were divided into 5 groups according to their possession of 0, 1, 2, 3 or > or = 4 components of the metabolic syndrome. The differences in all variables between groups were analyzed by ANOVA. The relationship between serum UA level and the number of metabolic components was determined by linear regression analysis. The contribution to elevated UA of possessing different risk factors was determined by a multivariate linear regression model. RESULTS: Mean serum UA level increased as the number of metabolic factors increased. Serum UA level was higher in subjects with abnormal triglyceride (TG), waist circumference, high-density lipoprotein cholesterol (HDL C) level and blood pressure (BP),with mean increases in UA level of 22.8, 21.4, 14.4 and 9.4 micromol/L, respectively (p < or = 0.001), compared to subjects with normal levels. After controlling for body mass index, abnormal TG, HDL-C and BP continued to account, in order of influence, for elevated UA. CONCLUSION: Serum UA level was elevated significantly as the number of metabolic components increased. Abnormal TG had the most influence on serum UA. A prospective study is warranted to determine if the prevention or treatment of hyperuricemia affects the development of metabolic syndrome. PMID- 17116613 TI - Different clinical presentations in chinese people with acute myocardial infarction in the emergency department. AB - BACKGROUND: The purpose of this study was to investigate the clinical characteristics of younger- ( 45 years) Chinese patients presenting to the emergency department (ED) with an initial acute myocardial infarction (AMI). METHODS: A retrospective review of 372 patients who had suffered an AMI (134 younger-aged, 238 older-aged), from 2,858 suspected AMI or ischemia patients during the period January 1996 to June 2003 inclusive, was conducted. Only patients who were diagnosed with AMI and who had been admitted to our institution's ED were enrolled into this study. RESULTS: The incidence of AMI for individuals who were < or = 45 years old was approximately 12.3% of those admitted to hospital under the impression of AMI. The percentage of males was more predominant in the younger-aged group (p < 0.01). Regarding major risk factors for coronary artery disease (CAD), younger patients were more likely to have a family history of cardiac disease (p < 0.01), obesity with an elevated body mass index (26.2 +/- 4.1 vs. 24.2 +/- 3.7, p < 0.01),and hyperlipidemia (p < 0.01) when compared to the older-aged patients, who were more likely to suffer from hypertension (p < 0.01) and/or diabetes mellitus than their younger-aged counterparts (p < 0.01). Younger patients also featured a higher incidence of single-vessel CAD (p = 0.05), an insignificant rate of CAD (p = 0.02), and a lower rate of triple-vessel CAD (p = 0.03). CONCLUSION: For Chinese, male gender and incidences of chest pain, positive family history of heart disease, obesity and hyperlipidemia were significantly greater in the younger aged AMI patient group than in the older-aged AMI patient group. Younger-aged patients also had a greater rate of single-vessel CAD but a lower rate of triple vessel CAD than older-aged patients. PMID- 17116614 TI - Health care utilization of home care patients at an academic medical center in Taiwan. AB - BACKGROUND: Previous surveys of home care patients in Taiwan have primarily concentrated on patients' status and needs. The aim of this study was to review the actual health care utilization of home care patients during the course of 1 year. METHODS: Home care patients at an academic medical center in Taiwan were selected and their insurance claims data at this hospital in 2001 were analyzed. Analyses included the patients' patterns and diagnoses of visits and admissions, and their drug utilization. For diagnoses made at outpatient departments, the grouping system from the National Hospital Ambulatory Medical Care Survey in the United States was used. The Anatomical Therapeutic Chemical Classification system was applied to drug grouping. RESULTS: The home care agency of the hospital cared for 165 patients (66 women, 99 men) in 2001. In total, these 165 patients received 1,358 home visits, 2,751 outpatient visits, and 108 inpatient admissions. While the most frequent diagnoses for all visits were cerebrovascular disease, hypertension, diabetes mellitus, chronic and unspecified bronchitis, psychoses, and other disorders of the central nervous system, the most frequent diagnoses at discharge from the hospital were urinary tract infection and pneumonia. In all visits, 12,282 items of drugs were prescribed in 2,337 prescriptions. On average, each prescription contained 5.3 +/- 2.8 items of drugs. The most frequently prescribed drugs were antacids, expectorants, laxatives, selective calcium channel blockers, and antithrombotic agents. CONCLUSION: The home care agency of the hospital should pay more attention to provision of comprehensive care and review of drug prescribing. PMID- 17116615 TI - Endoscopic sinus surgery under navigation system--analysis report of 79 cases. AB - BACKGROUND: Functional endoscopic sinus surgery (FESS) is the most popular method for treating medicine-refractory sinonasal disease. However, there are some pitfalls with this technique that can result in serious complications. Under the assistance of a navigation system, surgeons can overcome such problems. This study aimed to evaluate the efficacy of FESS aided by a navigation system. METHODS: There were 79 patients who underwent FESS under the assistance of a navigation system in our department between September 2004 and September 2005. Data on preoperative setup time, accuracy of the navigation system, operative time, and amount of blood loss during the operation were collected and analyzed. RESULTS: Mean preoperative setup time and mean operative time were 10.6 minutes and 112.3 minutes, respectively. The mean number of paranasal sinuses operated on was 5.8. The mean accuracy of the navigator system was 1.08 mm. Mean blood loss was 102.5 mL. Compared with data collected 2 years ago, preoperative setup time and operative time had both shortened. In addition, the number of operated paranasal sinuses had increased. This indicates that operative skill had improved. Moreover, operative time, amount of blood loss during the operation, and number of operated paranasal sinuses presented positive associations and significant differences (p < 0.05). No major complications such as blindness or cerebral spinal fluid leakage were noted. CONCLUSION: The characteristics of FESS aided by a navigation system include: (1) being able to pilot the relative positions of the operative instruments correctly in 3 dimensions; (2) being able to remove lesions more thoroughly; and (3) its inability to disclose the positions of vessels. As the number of patients we operated on increased, operative time was reduced. On the other hand, blood loss increased when the operations became more aggressive. However, performing FESS with the assistance of a navigation system is a safe way to treat patients with chronic paranasal sinusitis. PMID- 17116616 TI - Lactate dehydrogenase, not vascular endothelial growth factor or basic fibroblast growth factor, positively correlates to bone marrow vascularity in acute myeloid leukemia. AB - BACKGROUND: Angiogenesis has been extensively studied in acute myeloid leukemia (AML). Lactate dehydrogenase (LDH), a common biochemical marker for tumor burden and anaerobic glycolysis, is a poor prognostic factor for AML. Regulated by hypoxia-induced factor, both vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are responsive to cancer-related angiogenesis. To study the roles of serum LDH, VEGF and bFGF in AML angiogenesis, we investigated bone marrow vascularity in untreated AML patients, and analyzed its relationship to serum LDH, VEGF and bFGF levels. METHODS: Eighteen (11 males, 7 females; mean age, 57.7 years) de novo, untreated AML patients were enrolled. Bone marrow vascularity was evaluated by staining bone marrow core biopsy tissue with endothelial cell marker CD31 or CD34. Serum LDH was determined with the Wroblewski-La Due method. Serum VEGF and bFGF were determined with enzyme-linked immunoassay. The relationship of LDH, VEGF and bFGF level to bone marrow vessel numbers was examined by linear regression. RESULTS: Log LDH significantly correlated to AML bone marrow vascularity (r = 0.61; p = 0.007). VEGF and bFGF concentrations did not correlate with AML angiogenesis. CONCLUSION: These results suggest that serum LDH, but not VEGF and bFGF concentrations, can be used as a simple parameter for predicting vessel formation in AML bone marrow. PMID- 17116617 TI - Retroperitoneal bronchogenic cyst mimicking pancreatic cystic lesion. AB - Retroperitoneal bronchogenic cyst is detected extremely rarely and often masquerades as other diseases. Here, we report 2 cases of retroperitoneal bronchogenic cyst mimicking pancreatic mucinous tumor. Histologically, both cysts were composed of ciliated respiratory-like epithelium with abundant mucin content, smooth muscle bundles and mature cartilage, compatible with the diagnosis of retroperitoneal bronchogenic cyst. In addition to these 2 cases, another 42 retroperitoneal bronchogenic cysts reported in the English literature were collected for review and analysis. Twelve (28%) were located over the peripancreatic area. Just over half (51%) of them were asymptomatic. No accurate preoperative diagnosis could be made for any of the lesions. About a third (33.3%) of the peripancreatic retroperitoneal bronchogenic cysts masqueraded as pancreatic cystic lesions. PMID- 17116618 TI - Lumbosacral plexus injury and brachial plexus injury following prolonged compression. AB - We report the case of a 36-year-old woman who developed right upper and lower limb paralysis with sensory deficit after sedative drug overdose with prolonged immobilization. Due to the initial motor and sensory deficit pattern, brachial plexus injury or C8/T1 radiculopathy was suspected. Subsequent nerve conduction study/electromyography proved the lesion level to be brachial plexus. Painful swelling of the right buttock was suggestive of gluteal compartment syndrome. Elevation of serum creatine phosphokinase and urinary occult blood indicated rhabdomyolysis. The patient received medical treatment and rehabilitation; 2 years after the injury, her right upper and lower limb function had recovered nearly completely. As it is easy to develop complications such as muscle atrophy and joint contracture during the paralytic period of brachial plexopathy and lumbosacral plexopathy, early intervention with rehabilitation is necessary to ensure that the future limb function of the patient can be recovered. Our patient had suspected gluteal compartment syndrome that developed after prolonged compression, with the complication of concomitant lumbosacral plexus injury and brachial plexus injury, which is rarely reported in the literature. A satisfactory outcome was achieved with nonsurgical management. PMID- 17116619 TI - Clindamycin-induced anaphylactic shock during general anesthesia. AB - Clindamycin-related anaphylactic reaction is rarely reported. We report a male patient with buccal cancer who was undergoing radical neck dissection when life threatening anaphylactic shock developed soon after intravenous infusion of clindamycin. Immediate cardiopulmonary resuscitation was performed, and the patient recovered uneventfully. Perioperative anaphylactic shock is a serious problem due to the difficulty of judgment and potentially disastrous outcome. Immediate diagnosis and halting of drug infusion should be the first actions taken. PMID- 17116620 TI - Imatinib mesylate therapy in advanced gastrointestinal stromal tumors: experience from a single institute. AB - Gastrointestinal stromal tumors (GIST) are rare soft tissue sarcomas arising primarily from mesenchymal tissue in the gastrointestinal tract and abdomen. Since there is no effective treatment in the advanced stages, the outcome is poor in such patients. Recently, imatinib mesylate, a selective tyrosine kinase inhibitor, has shown a promising effect in GIST. Hence, we report our experience on the management of advanced GIST with imatinib therapy. A total of 14 patients were enrolled in this study, including 10 males and four females (median age, 51 years). The results showed that the small intestine was the most frequent site of primary lesion, while the liver was the most frequently metastasized organ. Most of the patients experienced tolerable side effects with imatinib therapy, including edema of periorbital area and/or legs and abdominal pain. Only two mortalities were noted during follow-up. The patients clinically benefited from imatinib therapy, with one patient having a complete response, three having a partial response, and seven having stable disease. The results demonstrate promising effects of imatinib in advanced GIST. PMID- 17116621 TI - Multivariate analysis of prognostic determinants for colorectal cancer patients with high preoperative serum CEA levels: prognostic value of postoperative serum CEA levels. AB - High preoperative serum carcinoembryonic antigen (CEA) levels have been well investigated and found to be associated with poor prognosis in patients with colorectal cancer (CRC). However, it has been observed that the outcome varies after curative resection, along with postoperative serum CEA levels; some patients continue to have high postoperative serum CEA levels while postoperative CEA levels return to normal in others. The purpose of this study was to determine the prognostic significance of postoperative serum CEA levels in CRC patients with high preoperative serum CEA levels. Between January 2002 and December 2004, 423 CRC patients underwent operation in our hospital; 181 (42.8%) had high preoperative serum CEA levels and were enrolled in this study. Among the 181 patients, 165 patients had curative resection; the remaining 16 had stage IV disease, so they underwent palliative surgery and were subsequently excluded from analysis. Pre- and postoperative serum CEA levels were measured and analyzed. All patients had curative resection and were divided into two groups according to postoperative serum CEA levels: one group comprised patients with postoperative serum CEA > or = 5 ng/mL (n = 80) and the other group comprised patients with postoperative serum CEA levels < 5 ng/mL (n = 85). Postoperative serum CEA levels were significantly related to location of primary tumors (p = 0.042), lymph node metastases (p = 0.009), TNM stage (p = 0.001), and postoperative relapse (p = 0.004). The results of multivariate analysis showed that both lymph node metastases and high postoperative serum CEA levels (> or = 5 ng/mL) were independent prognostic factors for CRC patients after curative resection. Postoperative serum CEA levels can be a single independent prognostic determinant in CRC patients with high preoperative serum CEA levels. Intensive follow-up and adjuvant therapy may be necessary in CRC patients who continue to have high postoperative serum CEA levels even after curative resection. PMID- 17116622 TI - Effect on spasticity after performance of dynamic-repeated-passive ankle joint motion exercise in chronic stroke patients. AB - Spasticity associated with abnormal muscle tone is a common motor disorder following stroke, and the spastic ankle may affect ambulatory function. The purpose of this study was to investigate the short-term effect of dynamic repeated-passive ankle movements with weight loading on ambulatory function and spastic hypertonia of chronic stroke patients. In this study, 12 chronic stroke patients with ankle spasticity and inefficient ambulatory ability were enrolled. Stretching of the plantar-flexors of the ankle in the standing position for 15 minutes was performed passively by a constant-speed and electrically powered device. The following evaluations were done before and immediately after the dynamic-repeated-passive ankle movements. Spastic hypertonia was assessed by the Modified Ashworth Scale (MAS; range, 0-4), Achilles tendon reflexes test (DTR; range, 0-4), and ankle clonus (range, 0-5). Improvement in ambulatory ability was determined by the timed up-and-go test (TUG), the 10-minute walking test, and cadence (steps/minute). In addition, subjective experience of the influence of ankle spasticity on ambulation was scored by visual analog scale (VAS). Subjective satisfaction with the therapeutic effect of spasticity reduction was evaluated by a five-point questionnaire (1 = very poor, 2 = poor, 3 = acceptable, 4 = good, 5 = very good). By comparison of the results before and after intervention, these 12 chronic stroke patients presented significant reduction in MAS and VAS for ankle spasticity, the time for TUG and 10-minute walking speed (p < 0.01). The cadence also increased significantly (p < 0.05). In addition, subjective satisfaction with the short-term therapeutic effect was mainly good (ranging from acceptable to very good). In conclusion, 15 minutes of dynamic repeated-passive ankle joint motion exercise with weight loading in the standing position by this simple constant-speed machine is effective in reducing ankle spasticity and improving ambulatory ability. PMID- 17116623 TI - Primary appendiceal malignancy: a clinicopathologic study. AB - Primary appendiceal cancer is a rare neoplasm. We analyzed the clinicopathologic characteristics and clinical outcomes of patients with primary appendiceal cancer treated at Buddhist Tzu Chi General Hospital in Eastern Taiwan. We reviewed the medical records of patients who had appendectomy at our hospital over a 10-year period and studied those who had histologically proven malignant appendiceal neoplasms. We treated eight such patients: seven males and one female. Their median age was 66 years (range, 59-78 years). There were three mucinous adenocarcinomas, two colonic type adenocarcinomas, and three adenocarcinoids. No patient was diagnosed correctly before surgery, and five (62.5%) had a preoperative diagnosis of acute appendicitis. Operative procedures included right hemicolectomy or partial colectomy in four, debulking and right hemicolectomy in three, and appendectomy only in one. With a mean follow-up of 64 months (range, 3 132), patients with adenocarcinoid lesions had better prognosis than those with adenocarcinomas. The important prognostic factors of primary appendiceal cancer included histologic subtypes and the extent of dissemination. In our series, palliative resection for disseminated lesions with or without additional chemotherapy resulted in long-term survival. PMID- 17116624 TI - Traumatic tricuspid insufficiency with chordae tendinae rupture: a case report and literature review. AB - With the increase in the number of automobile accidents, traumatic tricuspid insufficiency, a rare complication of non-penetrating blunt chest injury, has become an important problem. This kind of injury has been found more frequently during the last decade, partly because of better diagnostic procedures and a better understanding of the pathology. Here, we report a 22-year-old male patient who suffered chest trauma from an automobile accident. Echocardiography demonstrated tricuspid chordae tendinae rupture with remarkable tricuspid regurgitation. We discuss this case in comparison with the previous literature. This case reminds us that physicians in the emergency department should be aware of this potential complication following non-penetrating chest trauma. PMID- 17116625 TI - Postmalaria neurologic syndrome: a case report. AB - Neurologic signs and symptoms are common in acute malarial infection. However, after the parasites have been cleared from the blood and patients recover full consciousness, neurologic or psychiatric symptoms may occur or recur within 2 months after the acute illness. This phenomenon is called "postmalaria neurologic syndrome" (PMNS). We present a 50-year-old man who returned from the Republic of Malawi and soon developed Plasmodium falciparum malaria. Cerebral malaria, renal failure, hepatic failure, diffuse intravascular coagulation with thrombocytopenia, and upper gastrointestinal bleeding were noted during the acute stage. He was admitted to the infectious diseases ward and treated for 3 weeks. He was free from clinical general symptoms and parasites in blood smear when discharged. However, 2 weeks after discharge, he began to experience severe headache, dizziness, diplopia, mild hand tremor, unsteady gait, and easy falling. When readmitted to the neurologic ward, he presented with irritability, delirium, visual hallucination, and strange behavior. Neurologic examination was normal except for mild general weakness and evident truncal ataxia when walking. Brain magnetic resonance imaging revealed no structural lesions, and electroencephalography showed diffuse cortical dysfunction. Cerebral spinal fluid profile exhibited cytoalbuminologic dissociation. Brain single photon emission computed tomography showed diffuse cerebral parenchymal disorder. Nerve conduction studies revealed early sensory predominant polyneuropathy. The unsteadiness persisted for the initial 2 weeks of hospitalization until corticosteroid was administered. Intravenous methylprednisolone (80 mg/day) was continued for 3 days, followed by oral prednisolone (45 mg/day). His unsteadiness improved gradually after medication, and he absconded from the hospital on the 9th day of corticosteroid treatment with clear consciousness and free ambulation. The manifestation of PMNS is diverse and may present as an acute confusional state or psychosis, generalized seizure, fine tremors, cerebellar syndromes, postural hypotension, or malarial polyneuritis. Although the neurologic syndrome is primarily self-limited in most cases, corticosteroid may be beneficial in reversing PMNS. PMID- 17116626 TI - Tube thoracostomy-related necrotizing fasciitis: a case report. AB - Spontaneous pneumothorax is a serious complication of pulmonary tuberculosis that requires immediate treatment. Necrotizing fasciitis is a serious, rapidly progressive infection of the subcutaneous tissue and fascia, most related to trauma or surgery. Here, we report a case of pulmonary tuberculosis with spontaneous pneumothorax. A standard procedure of tube thoracostomy was performed for lung re-expansion. Two days after the tube was removed, necrotizing fasciitis developed from the puncture site. Computed tomography of the chest showed focal thickness with gas formation and loss of the fat plane over the chest wall, which is compatible with the diagnosis of necrotizing fasciitis. Aggressive treatment was given, including emergency fasciectomy and adequate systemic antibiotic and antituberculous treatment. The necrotizing fasciitis was successfully treated. The patient was discharged and sent home with maintenance antituberculous therapy. PMID- 17116627 TI - Subcutaneous emphysema and pneumomediastinum secondary to dental extraction: a case report and literature review. AB - Here, we present the case of a woman who suffered from acute dyspnea and right cheek and neck swelling during molar extraction. The use of a high-speed dental drill may introduce air into the soft tissue and lead to subcutaneous emphysema and pneumomediastinum. After a review of the literature, we found that subcutaneous emphysema and pneumomediastinum are rare complications secondary to dental extraction. We report this case because physicians in the emergency department may misdiagnose the symptoms as an allergic reaction. Dentists should be more aware of air leak during dental extraction. PMID- 17116628 TI - [Vector capacities of Similium damsnosum s.l and risk for Onchocerca volvulus transmission in Inga (Democratic Republic of Congo)]. AB - This article analyzes the results of a one-year entomological survey conducted near the Yalala rapids in the Inga region, Democratic Republic of Congo, prior to the implementation of Community-Directed Treatment with Ivermectin (CDTI). A vector control program had been undertaken from October 1969 to December 1980, 20 kilometers upstream from the study site. The results of the present study show that during 122 collection days, 5,824 females were captured, of which 3,899 were dissected; 2,001 (51.3%) were parous while 44 (2.2%) were infected while 4 (0.2%) were infective. The crude Annual Transmission Potential (ATP) was 65 infective larvae per man and per year at the river as against 12 in Yalala village located two kilometers from the rapids. The average biting rate was 5 flies/man/day during the dry season and 90 during the rainy season. The vector capacity of similium shows that the risk of transmission of Onchocerca volvulus to the human population was not only reduced in time (2-3 months), but was also negligible all year round. PMID- 17116629 TI - [Prevalence of glaucomas in a Black Cameroonian population]. AB - OBJECTIVES: To determine the prevalence and the various types of glaucoma in a Black urban Cameroonian population. MATERIAL AND METHOD: A retrospective analysis of 1,343 glaucomatous patients' files (757 men and 586 women) from a total of 24,462 Cameroonian patients of all ages was carried out from January 1991 to December 2001 in the Douala General Hospital in Cameroon in Central Africa. RESULTS: The prevalence of glaucoma of all types is 5.5%. It increases with age. Of subjects with glaucoma, 79.6% had not been previously diagnosed. The mean patient's age is 53.3 (+/-17.1) years. The mean intraocular pressure is 29.6 (+/ 12.6) mmHg. The mean vertical cup-disc ratio is 0.7 (+/-0.2). The prevalence of primary glaucoma of any type is 4.5%. The overall prevalence of primary open angle glaucoma, primary angle-closure glaucoma and congenital glaucoma was 4.3%, 0.03% and 0.07%, respectively. The normal-tension glaucoma is rare in this study (6.3% of primary open-angle glaucoma).The most common form of primary angle closure glaucoma is chronic angle closure glaucoma (61,1%) and is usually undiagnosed due to a lack of gonioscopic evaluation. The prevalence of secondary glaucoma of any type is 1%. The most common forms of secondary angle closure glaucoma were neovascular glaucoma (47.4% of cases with the prevalence of 0.5%) and post-traumatic glaucoma (23.1% of cases with the prevalence of 0.2%). Prevalence of bilateral and monocular blindness due to glaucoma is 8% (108/1,343) and 32.9% (441/1,343) respectively of glaucoma patients. CONCLUSION: This study confirms the high prevalence of glaucoma in a black African population. Gonioscopy is an important diagnostic tool that should be promoted to guide more effective glaucoma treatment in our region. The very high rate of blindness among the glaucoma patients in Cameroon is due to a lack of care. The patients are young, the disease is advanced, and compliance and follow-up are poor. Glaucoma surgery therefore offers a favourable option from an economical standpoint. Glaucoma is a devastating disease and constitutes a significant public health problem. PMID- 17116630 TI - [Torticollis in a group of strabismic patients in Cameroon]. AB - PURPOSE: Our aim was to study the characteristics of torticollis in a group of strabismic patients in Cameroon. PATIENTS AND METHODS: This was a prospective study carried out in the ophthalmology service of the General Hospital, Douala from January 1991 to December 2004. All strabismic patients followed up in our service were included in this study and they all had a complete strabologic and ophthalmological examination. Torticollis was analysed using photographs and classified according to its configuration, degree and variability. RESULTS: We examined 379 strabismic patients of which 227 were divergent (59.9%) and 152 convergent (40.1%). We found 166 cases of torticollis (43.8%). This pathology was found in 52.6% of patients with esotropia versus 37.9% in exotropia. Torticollis was classified as severe in 14.5% of patients, moderate in 45.8% and mild in 39.7%. Two out of three early esotropias had alternating torticollis. In congenital monophthalmus syndrome, torticollis was found in 68.4% of cases and in 100% of cases with the alphabetic syndrome. Moreover 63.8% of strabismics with torticollis have astigmatism. CONCLUSION: Torticollis in strabismic patients is a cause for concern as it is frequent and its functional consequences are severe. PMID- 17116631 TI - [Topography of ischemic strokes in Abidjan (Cote d'Ivoire): a computed tomographic approach]. AB - OBJECTIVE: The authors used computed tomography (CT) to assess and categorize the topography of ischemic strokes (IS) among blacks living in Abidjan, the commercial and administrative center of Cote d'Ivoire, in West Africa. METHODS: This retrospective study analyzed CT data of patients admitted to the Sainte Anne Marie Polyclinic (the principal private hospital in the country) and to the neurology department of the university hospital center in Cocody, from January 1, 2000, to December 31, 2001. The study included patients who met World Health Organisation criteria for stroke and had CT performed during the hospitalization for this stroke. We examined CT data to find early and late signs of IS, analyze lesion diameter (15 mm cutoff used to distinguish infarcts from lacuna), and determine their topography (cerebral arterial territory and localization, that is, brain lobes, basal ganglia and posterior cerebral fossa). RESULTS: We included 260 subjects (58% males) with a median age of 45 years (range: 20-80 years). CT findings were abnormal for 224 patients with infarcts (72.7%), lacuna (27.3%), or both (8%). As reported elsewhere, the anterior arterial territory was most often affected (83.9%) with a middle cerebral artery lesion in 79.4% of patients. Posterior territory (16.1%) lesions and lacuna were probably underestimated because CT exploration is reported to be less useful for this area than for the carotid area. On the other hand, CT diagnoses infarcts more easily than it does lacuna. CT was normal for 36 patients although performed no more than 3 days after IS. These patients did not undergo CT angiography, which might have shown cerebral artery occlusion. CONCLUSION: Our study included IS of all types and typography. Stroke registries in Africa would provide useful data for better assessment of prevalence for specific topographic and etiologic types of stroke. PMID- 17116632 TI - [Assessment of 24 years of infant growth monitoring in a rural area of Senegal (1969-1992)]. AB - Undernutrition in young children in developing countries is associated with an increased risk of death. But in several studies, a decrease in mortality was not associated with any decrease in the prevalence of undernutrition. STUDY AREA: A rural population of Casamance (Senegal) has been under yearly demographic surveillance by The French National Institute of Demographic Studies (INED) since 1985. Between 1960-1964 and 1990-1994, under-5 and child (1-4 years) mortality rates dropped from 312 to 127 and from 201 to 68 per 1,000, respectively. Since 1961, French Catholic nuns who are also professional nurses have been in charge of a private village dispensary located in a rural area of Casamance. This dispensary delivers permanent, high quality service and is widely attended. GROWTH MONITORING PROGRAMME: A growth-monitoring programme, supported by Cathwell, was initiated in 1969 for 0-5-year-old children (0-3 from 1985). Children were weighed wearing light clothes to the nearest 10g. Weights were recorded in a register that also contained information concerning identity (name, sex, date of birth) and address. All weights were plotted on growth charts kept by the mothers. During the sessions, the nurses provided nutrition education messages (i.e., preparation of high-energy and nutrient-dense infant gruels using local foods), advice on illness management (oral rehydration during diarrhoea) and hygiene (well and water-jar disinfection, construction of pit-latrines), importance of vaccination. From 1975, they also distributed free chloroquine during the malaria transmission season (May-November) for prevention and early presumptive treatment. Most likely thanks to this programme, infant and child mortality reached a low level at the end of the 1980s. In 1990, plasmodium resistance to chloroquine appeared, increasing malaria mortality. METHODS: All weight measurements taken in 1969-1994 were entered into a database. This paper presents an analysis of weight measurements taken at 3-23 months of age from 1969 1992. A total of 4,636 infants were weighed at least once, but only 3,912 infants (1,983 boys and 1,929 girls) were available for the analysis, 724 being excluded due to missing data. RESULTS: The average coverage of the programme during the month of February was 88% for infants aged 3-23 months. Mean weight was examined at three target ages: 5, 11 and 15 months. Not only did the nutritional status not improve between 1969 and 1989, it even deteriorated in some years for all three age-cohorts. The nutritional status of infants in this community did not differ significantly from that of 12-23 month-old Senegalese children in the 1992 1993 Demographic and Health Survey (DHS). Seasonal differences in mean weight and the prevalence of underweight became significant in the rainy season since 1975. Underweight for the 15-month-old children increased over time during the rainy season. These findings were unexpected, since malaria morbidity is thought to be at least partially responsible for seasonal variations in the nutritional status of young children, and despite the fact that the nurses began a malaria control programme in 1975. CONCLUSION: The rapid transition towards lower childhood mortality observed in this rural area of Casamance (Senegal), was not concomitant with any improvement in infants' nutritional status from 1969 through 1992. Focused public health interventions such as vaccinations and malaria prevention probably did not improve the nutritional status. Paradoxically, growth monitoring may have been more helpful in improving health than growth. Targeted specific nutritional interventions are needed to significantly improve growth of children in this community. PMID- 17116633 TI - [Low birth weight in Maniema (Democratic Republic of Congo)]. AB - BACKGROUND: This study sought to examine birth outcomes in the general population in the Democratic Republic of Congo (DRC). Specifically we wanted to see if the proportion of low birth-weight infants (<2,500g), including preterm infants, differed from that of most developing countries in view of the political disturbances in DRC over the past eight years. METHODS: We studied birth weight in two maternity units in the province of Maniema from November 2003 to October 2004: there were 938 births, 450 at Kama and 488 at Kipaka. RESULTS: The rate of low birth weight (LBW) (<2,500g) infants was 164 per 1,000 live births at Kama and 270 per 1,000 live births at Kipaka. This rate was higher at Kipaka than at Kama, probably because commerce has improved living conditions at Kama (chi(2)=16.79, p<0.001). The study also confirmed the importance of maternal age, especially the 14-17 year age group, in low birth weight (including preterm births). The relative risk associated with this age group was 2.2 at Kama (95% CI: 1.3-3.7) and 1.5 at Kipaka (95% CI: 1.1-2.1). CONCLUSION: These results suggest that low birth weight (including preterm birth) remains a public heath problem in DRC, as in other developing countries. They also confirm that very young mothers are most likely to have low birth-weight infants and they stress the importance of resources for preventing preterm birth and improving care of low birth-weight infants, born before or at term. PMID- 17116634 TI - [Management of laryngeal cancer in an ENT Service in subSaharan Africa]. AB - INTRODUCTION: The management of cancers of the larynx remains a challenge to most ENT specialists in Cameroon due to a multitude of factors linked to their management. The aim of this study is to identify the important factors that militate against the proper management of these cancers and to propose solutions. MATERIALS AND METHODS: A retrospective study was carried out in the ENT service of the General Hospital of Douala over six years (April 1998 to June 2006). Eighteen cases of epidermoid cancers of the larynx were identified. Only 10 of the clinical records could be reviewed because the remaining patients refused to be treated in our unit immediately after they were informed of the diagnosis. RESULTS: All the cases were males with a mean age of 53. Most of the cases diagnosed were at the advanced stage of the disease (7T4, 2T3, 1T2). Six patients were treated with radiotherapy only, one with chemo-radiotherapy. Three patients had a full laryngectomy followed by radiation therapy in two cases. One of the cases operated developed a pharyngeal fistula. By the end of the third year only 30% of our patients still continued their follow-up. DISCUSSION: The therapeutic protocols used took into consideration the socio-economic status of the patients, the experience of the personnel in the management of laryngeal cancers and the treatment techniques and modes available in our hospital. Because of these factors, standard European and American recommendations for the management of laryngeal cancers could not be used. CONCLUSION: Government intervention to offer subventions for the high cost of work-up and treatment of these patients is a must for better results to be obtained. There is also a need for the creation of training, prevention and treatment centres for cancers of the larynx. PMID- 17116635 TI - ["Street" medication in Burkina Faso: local names, social relationships, and alleged therapeutic effects]. AB - This qualitative assessment, based on discussions and discourse collected in interviews with members of the general population, addresses the popular view of pharmaceutical drugs in Burkina Faso. The main results demonstrate a strong preference for drugs sold in the street and their largely "off-label" uses. These drugs not only treat defined diseases but also generate street discussions, popular images and social relationships that lead to their consumption, sometimes excessive. Furthermore, the links between the legal and illegal street markets, related in part to the legal status of different drugs, also leads to questions about good and bad, true and false. These distinctions, considered as labels, influence the population's behavior and attitude concerning street medication. PMID- 17116636 TI - [Factors associated with adherence to therapy among HIV-infected patients in Bangui]. AB - OBJECTIVE: To identify the factors related to good or non-adherence met among patients under highly active antiretroviral therapy (HAART) or cotrimoxazole prophylaxis (CTX) in Bangui. METHOD: A cross-sectional study was conducted on a sample representative of patients with HIV followed up in two health centres in Bangui. Patients had been under treatment for at least 2 months. Information concerning adherence was collected through a questionnaire and by interview of the patients. The questionnaire was designed to measure if patients with HIV were taking less than the total number of antiretroviral or CTX pills prescribed by their physician. Adherence was measured by the patient self-reported question of taking the drugs during the last 4 days and the remaining pill count (RPC). RESULTS: 141 patients with the criteria of inclusion were questioned; among these, 89 patients under antiretroviral treatment (d4T/3TC/NVP) and 52 patients under cotrimoxazole. In the study, adherence varies from 67.3 to 94.3 %. According to the type of treatment, adherence of the patients under CTX is lower (65.4 %) than that of the patients under HAART (77.5 %). Adherence was better for the patients who had a personal project and with a need of information on HIV infection. It is worse for the patients with another affection. CONCLUSION: These results suggest the necessity of assessing and supporting HIV-infected adult's adherence routinely in AIDS care institutions. In addition, the method used to assess adherence must be simple, accessible and low-cost for the countries with limited resources. PMID- 17116637 TI - [Facial entomophtoromycosis: a propos of a case]. AB - We report one case of rare facial entomophtoromycosis which occured in a young male patient of 16 years of age, in Cameroon. The clinical presentation was a central mass of the face. We show the interest of the diagnosis of this rare fungal infection through biopsy and its medical treatment. PMID- 17116638 TI - ENU mutagenesis identifies mice with cardiac fibrosis and hepatic steatosis caused by a mutation in the mitochondrial trifunctional protein beta-subunit. AB - Using the metabolomics-guided screening coupled to N-ethyl-N-nitrosourea-mediated mutagenesis, we identified mice that exhibited elevated levels of long-chain acylcarnitines. Whole genome homozygosity mapping with 262 SNP markers mapped the disease gene to chromosome 5 where candidate genes Hadha and Hadhb, encoding the mitochondria trifunctional protein (MTP) alpha- and beta-subunits, respectively, are located. Direct sequencing revealed a normal alpha-subunit, but detected a nucleotide T-to-A transversion in exon 14 (c.1210T>A) of beta-subunit (Hadhb) which resulted in a missense mutation of methionine to lysine (M404K). Western blot analysis showed a significant reduction of both the alpha- and beta subunits, consistent with reduced enzyme activity in both the long-chain 3 hydroxyacyl-CoA dehydrogenase and the long-chain 3-ketoacyl-CoA thiolase activities. These mice had a decreased weight gain and cardiac arrhythmias which manifested from a prolonged PR interval to a complete atrio-ventricular dissociation, and died suddenly between 9 and 16 months of age. Histopathological studies showed multifocal cardiac fibrosis and hepatic steatosis. This mouse model will be useful to further investigate the mechanisms underlying arrhythmogenesis relating to lipotoxic cardiomyopathy and to investigate pathophysiology and treatment strategies for human MTP deficiency. PMID- 17116639 TI - Genome-wide SNP assay reveals structural genomic variation, extended homozygosity and cell-line induced alterations in normal individuals. AB - The recent hapmap effort has placed focus on the application of genome-wide SNP analysis to assess the contribution of genetic variability, particularly SNPs, to traits such as disease. Here, we describe the utility of genome-wide SNP analysis in the direct detection of extended homozygosity and structural genomic variation. We use this approach to assess the frequency of genomic alterations resulting from the lymphoblast immortalization and culture processes commonly used in cell repositories. We have assayed 408 804 SNPs in 276 DNA samples extracted from Epstein-Barr virus immortalized cell lines, which were derived from lymphocytes of elderly neurologically normal subjects. These data reveal extended homozygosity (contiguous tracts >5 Mb) in 9.5% (26/272) and 340 structural genomic alterations in 182 (66.9%) DNA samples assessed, 66% of which did not overlap with previously described structural variations. Examination of DNA extracted directly from the blood of 30 of these subjects confirmed all examined instances of extended homozygosity (6/6), 75% of structural genomic alteration <5 Mb in size (12/16) and 13% (1/8) of structural genomic alteration >5 Mb in size. These data suggest that structural genomic variation is a common phenomenon in the general population. While a proportion of this variability may be caused or its relative abundance altered by the immortalization and clonal process this will have only a minor effect on genotype and allele frequencies in a large cohort. It is likely that this powerful methodology will augment existing techniques in the identification of chromosomal abnormalities. PMID- 17116640 TI - Pael receptor induces death of dopaminergic neurons in the substantia nigra via endoplasmic reticulum stress and dopamine toxicity, which is enhanced under condition of parkin inactivation. AB - Selective loss of dopaminergic neurons is the final common pathway in Parkinson's disease. Expression of Parkin associated endothelin-receptor like receptor (Pael R) in mouse brain was achieved by injecting adenoviral vectors carrying a modified neuron-specific promoter and Cre recombinase into the striatum. Upregulation of Pael-R in the substantia nigra pars compacta of mice by retrograde infection induced endoplasmic reticulum (ER) stress leads to death of dopaminergic neurons. The role of ER stress in dopaminergic neuronal vulnerability was highlighted by their decreased survival in mice deficient in the ubiquitin-protein ligase Parkin and the ER chaperone ORP150 (150 kDa oxygen regulated protein). Dopamine-related toxicity was also a key factor, as a dopamine synthesis inhibitor blocked neuronal death in parkin null mice. These data suggest a model in which ER- and dopamine-related stress are major contributors to decreased viability of dopaminergic neurons in a setting relevant to Parkinson's disease. PMID- 17116641 TI - Disseminating sexually transmitted infections diagnostics information: the SDI web publication review series. AB - OBJECTIVES: The World Health Organization Sexually Transmitted Diseases Diagnostics Initiative (SDI) website publication review seeks to provide health care providers in all geographic and economic settings with timely, critical, and concise information concerning new developments in laboratory and field diagnosis of sexually transmitted infections (STI). METHODS: Since 2003, the website (www.who.int/std_diagnostics/literature_reviews) has disseminated information in the form of annotated abstracts and commentaries on articles covering studies of STI laboratory-based and rapid assays that are commercially available or under development. Articles identified through searches of PubMed, specific journals, and by referrals from Editorial Board members are selected for inclusion if they meet pre-specified criteria. The objectives, methods, results, and conclusions for each article are summarised and board members are invited to prepare commentaries addressing study design and applicability of findings to end users. RESULTS: Currently, 91 STI diagnostics experts from 17 countries on six continents serve on the Editorial Board. Twelve quarterly issues have been posted that include summaries of 214 original and 17 review articles published from January 2002 through March 2005, with expert commentaries on 153 articles. Interest in the site has increased every year. In 2005, over 36 700 unique visitors from more than 100 countries viewed over 75,000 pages of information. CONCLUSIONS: The SDI Publication Review series has the potential to contribute to SDI's goal of improving care for patients with STI by increasing knowledge and awareness of STI diagnostics. Given the proliferation of internet-based STI testing services, this website may be broadened to meet the needs of a wider range of users. PMID- 17116642 TI - Utility of the Determine Syphilis TP rapid test in commercial sex venues in Peru. AB - OBJECTIVES: This study sought to evaluate the utility of the Determine Syphilis TP test performed in Peruvian commercial sex venues for the detection of active syphilis; and determine the feasibility of integrating rapid syphilis testing for female sex workers (FSW) into existing health outreach services. METHODS: We tested 3586 female sex workers for syphilis by Determine in the field using whole blood fingerstick, and by rapid plasma reagin (RPR) and Treponema pallidum haemagglutination assay (TPHA) in a central laboratory in Lima using sera. RESULTS: 97.4% of the FSW offered rapid syphilis testing participated; and among those who tested positive, 87% visited the local health centre for treatment. More than twice as many specimens were RPR reactive using serum in Lima (5.7%) than tested positive by whole blood Determine in the field (2.8%), and although most were confirmed by TPHA, only a small proportion (0.7%) were RPR reactive at >or=1:8 dilutions, and likely indicating active syphilis. Sensitivity, specificity and positive predictive value of the Determine Syphilis TP test in whole blood when compared to serum RPR reactivity at any dilution confirmed by TPHA as the gold standard were 39.3%, 99.2% and 71.4%, respectively. Sensitivity improved to 64.0% when using serum RPR >or=1:8 confirmed by TPHA. Invalid tests were rare (0.3%). CONCLUSIONS: Rapid syphilis testing in sex work venues proved feasible, but Determine using whole blood obtained by fingerstick was substantially less sensitive than reported in previous laboratory-based studies using serum. Although easy to perform in outreach venues, the utility of this rapid syphilis test was relatively low in settings where a large proportion of the targeted population has been previously tested and treated. PMID- 17116643 TI - Opposite immune functions of GM-CSF administered as vaccine adjuvant in cancer patients. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) has been and is still widely used as an adjuvant in clinical trials of vaccination with autologous tumor cells, peptides and/or dendritic cells in a variety of human neoplasms. This cytokine was administered either as product of gene-transduced tumor cells or as recombinant protein together with the vaccine given subcutaneously or intradermally. Results of these trials were heterogeneous in terms of induction of vaccine-specific immune response and of clinical response. Though in some of these studies GM-CSF appeared to help in generating an immune response, in others no effect or even a suppressive effect was reported. Here, we review the literature dealing with the immune adjuvant activity of GM-CSF both in animal models and clinical trials. As a consequence of such analysis, we conclude that GM-CSF may increase the vaccine-induced immune response when administered repeatedly at relatively low doses (range 40-80 microg for 1-5 days) whereas an opposite effect was often reported at dosages of 100-500 microg. The potential mechanisms of the GM-CSF-mediated immune suppression are discussed at the light of studies describing the activation and expansion of myeloid suppressor cells by endogenous tumor-derived or exogenous GM-CSF. PMID- 17116644 TI - Disulfide-bond reshuffling in the evolution of an ape placental ribonuclease. AB - Disulfide bonds play important roles in the folding and stability of proteins and are evolutionarily conserved. A classic example is RNase A (also known as bovine pancreatic ribonuclease), which contains 4 conserved disulfide bonds among 8 cysteines. However, human RNase 8, a paralog of RNase A uniquely expressed in the placenta, has lost one of the conserved cysteines but gained another, when compared with RNase 8 of various monkeys and with RNase A. We here show that both the loss and gain of the cysteines in human RNase 8 occurred in the common ancestor of African great apes (humans, chimps, and gorillas) 7-13 MYA. Computational predictions suggest changes of disulfide bonding by these cysteine substitutions. Site-directed mutagenesis indicates that if the ribonucleolytic activity is essential for RNase 8's function, the gain of the cysteine must have preceded the loss. Human RNase 8 represents one of the first examples in which the presumable evolutionary change of a disulfide bond involves 1 loss and 1 gain of cysteine, instead of 2 losses or 2 gains. Our results provide the foundation for detailed analysis toward understanding the impact of disulfide-bond reshuffling on the structure, function, and evolution of proteins in general and human RNase 8 in particular. PMID- 17116645 TI - Cellular toxicity induced by SRF-mediated transcriptional squelching. AB - The transcriptional activator serum response factor (SRF) is a member of the immediate early gene family known to promote embryonic development, cell growth, and myogenesis through interaction with multiple nuclear protein factors. Previous studies have shown that SRF possesses potent transcriptional activation domains that can interfere with gene expression at artificially high expression levels through "transcriptional squelching." The current work sought to characterize toxicological aspects of SRF-mediated transcriptional squelching. An adenoviral expression system driven by the potent cytomegalovirus promoter was used to achieve up to a 50-fold increase in SRF protein levels. The overexpressed SRF is nuclear localized and interferes with gene expression independent of specific promoter interaction as expected for transcriptional squelching. SRF mediated squelching elicits robust cell killing affecting multiple cell types including normal and abnormal proliferating cells as well as postmitotic cells such as cardiomyocytes in culture, and the cell killing is more pronounced than that mediated by the tumor suppressor protein p53. Although both the DNA-binding and transcriptional activation domains of SRF are normally required for the physiological roles of SRF, only the transcriptional activation domain is required for cell killing. Unlike c-myc-induced cell killing, squelching-induced cell death does not require serum withdrawal and cannot be effectively attenuated by blocking the caspase and calpain proteolytic pathways or by overexpression of the antiapoptotic gene bcl-xL. These findings suggest transcriptional squelching may be engineered for killing cancer cells, and the SRF gene may represent a novel molecular target for cancer therapeutics. PMID- 17116646 TI - Computational methodologies for modelling, analysis and simulation of signalling networks. AB - This article is a critical review of computational techniques used to model, analyse and simulate signalling networks. We propose a conceptual framework, and discuss the role of signalling networks in three major areas: signal transduction, cellular rhythms and cell-to-cell communication. In order to avoid an overly abstract and general discussion, we focus on three case studies in the areas of receptor signalling and kinase cascades, cell-cycle regulation and wound healing. We report on a variety of modelling techniques and associated tools, in addition to the traditional approach based on ordinary differential equations (ODEs), which provide a range of descriptive and analytical powers. As the field matures, we expect a wider uptake of these alternative approaches for several reasons, including the need to take into account low protein copy numbers and noise and the great complexity of cellular organisation. An advantage offered by many of these alternative techniques, which have their origins in computing science, is the ability to perform sophisticated model analysis which can better relate predicted behaviour and observations. PMID- 17116647 TI - Marked hypercalcaemia in sepsis-induced multiple organ failure. PMID- 17116648 TI - Pesticide exposure and self-reported Parkinson's disease in the agricultural health study. AB - Previous studies based on limited exposure assessment have suggested that Parkinson's disease (PD) is associated with pesticide exposure. The authors used data obtained from licensed private pesticide applicators and spouses participating in the Agricultural Health Study to evaluate the relation of self reported PD to pesticide exposure. Cohort members, who were enrolled in 1993 1997, provided detailed information on lifetime pesticide use. At follow-up in 1999-2003, 68% of the cohort was interviewed. Cases were defined as participants who reported physician-diagnosed PD at enrollment (prevalent cases, n = 83) or follow-up (incident cases, n = 78). Cases were compared with cohort members who did not report PD (n = 79,557 at enrollment and n = 55,931 at follow-up). Incident PD was associated with cumulative days of pesticide use at enrollment (for highest quartile vs. lowest, odds ratio (OR) = 2.3, 95% confidence interval: 1.2, 4.5; p-trend = 0.009), with personally applying pesticides more than half the time (OR = 1.9, 95% confidence interval: 0.7, 4.7), and with some specific pesticides (ORs > or = 1.4). Prevalent PD was not associated with overall pesticide use. This study suggests that exposure to certain pesticides may increase PD risk. Findings for specific chemicals may provide fruitful leads for further investigation. PMID- 17116649 TI - Estimating lifetime risk of developing high serum total cholesterol: adjustment for baseline prevalence and single-occasion measurements. AB - The lifetime risk statistic is a powerful tool in epidemiology. It has been successfully applied to estimate and highlight the risks of numerous diseases, including breast cancer, Alzheimer's disease, stroke, and coronary heart disease and some of its risk factors. Application of this method to health-related conditions that may have an onset early in young adulthood or to measurements that can fluctuate over time introduces problems of under- or overestimation of risk. To correctly quantify the long-term risk of developing high serum total cholesterol (> or =240 mg/dl or use of lipid-lowering medication), the authors propose a key modification of the lifetime risk statistic: adjustment for baseline prevalence. It accounts for the fact that many people already have the condition at a young age (an age often chosen as baseline). The authors derive point estimators and confidence intervals and supply a SAS macro (SAS Institute, Inc., Cary, North Carolina). For assessment of the risk inflation due to single occasion measurement, the authors suggest two diagnostic tools, one requiring the condition to be present on two consecutive occasions and the other taking into account intrasubject variability. As an illustration, the authors calculate risk estimates for US Caucasians based on hypercholesterolemia incidence (1971-early 2001) from the Framingham Heart Study and prevalence data from the 1999-2000 National Health and Nutrition Examination Survey. PMID- 17116650 TI - Effects of past and recent blood pressure and cholesterol level on coronary heart disease and stroke mortality, accounting for measurement error. AB - The authors aimed to quantify the effects of current systolic blood pressure (SBP) and serum total cholesterol on the risk of mortality in comparison with SBP or serum cholesterol 25 years previously, taking measurement error into account. The authors reanalyzed 35-year follow-up data on mortality due to coronary heart disease and stroke among subjects aged 65 years or more from nine cohorts of the Seven Countries Study. The two-step method of Tsiatis et al. (J Am Stat Assoc 1995;90:27-37) was used to adjust for regression dilution bias, and results were compared with those obtained using more commonly applied methods of adjustment for regression dilution bias. It was found that the commonly used univariate adjustment for regression dilution bias overestimates the effects of both SBP and cholesterol compared with multivariate methods. Also, the two-step method makes better use of the information available, resulting in smaller confidence intervals. Results comparing recent and past exposure indicated that past SBP is more important than recent SBP in terms of its effect on coronary heart disease mortality, while both recent and past values seem to be important for effects of cholesterol on coronary heart disease mortality and effects of SBP on stroke mortality. Associations between serum cholesterol concentration and risk of stroke mortality are weak. PMID- 17116651 TI - Sex differences in semantic processing: event-related brain potentials distinguish between lower and higher order semantic analysis during word reading. AB - Behavioral studies suggest that women and men differ in the strategic elaboration of verbally encoded information especially in the absence of external task demand. However, measuring such covert processing requires other than behavioral data. The present study used event-related potentials to compare sexes in lower and higher order semantic processing during the passive reading of semantically related and unrelated word pairs. Women and men showed the same early context effect in the P1-N1 transition period. This finding indicates that the initial lexical-semantic access is similar in men and women. In contrast, sexes differed in higher order semantic processing. Women showed an earlier and longer lasting context effect in the N400 accompanied by larger signal strength in temporal networks similarly recruited by men and women. The results suggest that women spontaneously conduct a deeper semantic analysis. This leads to faster processing of related words in the active neural networks as reflected in a shorter stability of the N400 map in women. Taken together, the findings demonstrate that there is a selective sex difference in the controlled semantic analysis during passive word reading that is not reflected in different functional organization but in the depth of processing. PMID- 17116652 TI - Dynamic properties of excitatory synaptic connections involving layer 4 pyramidal cells in adult rat and cat neocortex. AB - To investigate the properties of excitatory connections between layer 4 pyramidal cells and whether these differed between rat and cat, paired intracellular recordings were made with biocytin filling in slices of adult neocortex. These connections were also compared with those from layer 4 spiny cells to layer 3 pyramids and connections between layer 3 pyramids. Connectivity ratios for layer 4 pyramid-pyramid pairs (1:14 cat, 1:18 rat) appeared lower than for the other types of connections studied in parallel, but excitatory postsynaptic potential (EPSP) amplitudes and time course were not significantly different either between species or across types of connection. Layer 4 pyramids targeted postsynaptic basal dendrites in both species, whether the pyramidal target was in layer 4 or layer 3. Within layer 4, relationships between mean EPSP amplitude, numbers of putative contacts, and distance between connected pairs indicated a rapid decline in connectivity strength with distance, equivalent to 3.4 mV and 10 synapses per 100 microm separation, from a maximum of 4 mV and 10 synapses at 0 microm. However, a subset, of burst-firing layer 4 pyramids, appeared to make no connections with other layer 4 spiny cells. Second EPSPs were depressed by 36% in rat and 28% in cat relative to first EPSPs at interspike intervals <15 ms. Subsequent EPSPs in brief trains were further depressed. Depression was predominantly presynaptic in origin. Recovery from depression could not be described adequately by a simple exponential for individual connections; it included peaks and troughs with periodicities of 10-15 ms. Complex relationships between the first 2 interspike intervals and third EPSP amplitude were also apparent in all connections so studied. Large third EPSPs followed specific combinations of first and second interspike intervals so that increasing, or decreasing, one without changing the other resulted in a smaller third EPSP. Finally, the outputs of layer 4 spiny cells to layer 3 exhibited partial recovery from depression during longer high-frequency trains, a property not apparent in the other connections studied. PMID- 17116653 TI - Endophenotypic studies in schizophrenia: promise and challenges. PMID- 17116654 TI - Thromboangiitis obliterans or Buerger's disease: challenges for the rheumatologist. AB - Thromboangiitis obliterans (Buerger's disease)--a type of vasculitis in young, mostly male subjects--remains strangely linked to smoking, which determines its occurrence, progression and prognosis by currently unknown mechanisms. It affects the small and medium-sized arteries and veins of the limbs. Despite the usual absence of systemic signs and symptoms, initially intermittent arthritis, even if not observed in all cases, confer on this disease the status of a systemic vasculitis. Diagnosis requires the elimination of many other diseases. The severity of the disease lies in the need for amputation in more than a quarter of all patients. Complete cessation of smoking remains the cornerstone of therapy. Local care is the second essential element of treatment. Prostacycline analogues can be used to help the patient through critical ischaemia. Blockade with antagonists of cannabinoid or endothelin receptors and the use of gene- or cell based therapy to induce therapeutic angiogenesis have opened up new possibilities for treatment. PMID- 17116655 TI - "To do no harm". PMID- 17116656 TI - Assessment of digital vascular structure and function in response to bosentan in patients with systemic sclerosis-related Raynaud's phenomenon. PMID- 17116657 TI - Alkaptonuria: a 60-yr follow-up. PMID- 17116658 TI - The C-terminal domains of vertebrate CstF-64 and its yeast orthologue Rna15 form a new structure critical for mRNA 3'-end processing. AB - Yeast Rna15 and its vertebrate orthologue CstF-64 play critical roles in mRNA 3 ' end processing and in transcription termination downstream of poly(A) sites. These proteins contain N-terminal domains that recognize the poly(A) site, but little is known about their highly conserved C-terminal regions. Here we show by NMR that the C-terminal domains of CstF-64 and Rna15 fold into a three-helix bundle with an uncommon topological arrangement. The structure defines a cluster of evolutionary conserved yet exposed residues we show to be essential for the interaction between Pcf11 and Rna15. Furthermore, we demonstrate that this interaction is critical for the function of Rna15 in 3 '-end processing but dispensable for transcription termination. The C-terminal domain of the Rna15 homologue Pti1 contains critical sequence alterations within this region that are predicted to prevent Pcf11 interaction, providing an explanation for the distinct functions of these two closely related proteins in the 3 '-end formation of RNA polymerase II transcripts. These results define the role of the C-terminal half of Rna15 and provide insight into the network of protein/protein interactions responsible for assembly of the 3 '-end processing apparatus. PMID- 17116659 TI - Signaling of mitochondrial biogenesis following oxidant injury. AB - Mitochondrial dysfunction is a common consequence of ischemia-reperfusion and drug injuries. For example, sublethal injury of renal proximal tubular cells (RPTCs) with the model oxidant tert-butylhydroperoxide (TBHP) causes mitochondrial injury that recovers over the course of six days. Although regeneration of mitochondrial function is integral to cell repair and function, the signaling pathway of mitochondrial biogenesis following oxidant injury has not been examined. A 10-fold overexpression of the mitochondrial biogenesis regulator PPAR-gamma cofactor-1alpha (PGC-1alpha) in control RPTCs resulted in a 52% increase in mitochondrial number, a 27% increase in respiratory capacity, and a 30% increase in mitochondrial protein markers, demonstrating that PGC-1alpha mediates mitochondrial biogenesis in RPTCs. RPTCs sublethally injured with TBHP exhibited a 50% decrease in mitochondrial function and increased mitochondrial autophagy. Compared with the controls, PGC-1alpha levels increased 12-fold on days 1, 2, and 3 post-injury and returned to base line on day 4 as mitochondrial function returned. Inhibition p38 MAPK blocked the up-regulation of PGC-1alpha following oxidant injury, whereas inhibition of calcium-calmodulin-dependent protein kinase, calcineurin A, nitric-oxide synthase, and phosphoinositol 3 kinase had no effect. The epidermal growth factor receptor (EGFR) was activated following TBHP exposure, and the EGFR inhibitor AG1478 blocked the up-regulation of PGC-1alpha. Additional inhibitor studies revealed that the sequential activation of Src, p38 MAPK, EGFR, and p38 MAPK regulate the expression of PGC 1alpha following oxidant injury. In contrast, although Akt was activated following oxidant injury, it did not play a role in PGC-1alpha expression. We suggest that mitochondrial biogenesis following oxidant injury is mediated by p38 and EGFR activation of PGC-1alpha. PMID- 17116660 TI - First characterization of fluoroquinolone resistance in Streptococcus suis. AB - We have identified and sequenced the genes encoding the quinolone-resistance determining region (QRDR) of ParC and GyrA in fluoroquinolone-susceptible and resistant Streptococcus suis clinical isolates. Resistance is the consequence of single point mutations in the QRDRs of ParC and GyrA and is not due to clonal spread of resistant strains or horizontal gene transfer with other bacteria. PMID- 17116661 TI - Quantitative immunoassay to measure plasma and intracellular atazanavir levels: analysis of drug accumulation in cultured T cells. AB - We have developed an enzyme immunoassay to measure atazanavir (ATV) levels in plasma and cells. Anti-ATV polyclonal antibodies were raised in rabbits by using a synthetic ATV derivative coupled to bovine serum albumin as the immunogen, and the enzyme tracer was prepared by chemically coupling the ATV derivative with acetylcholinesterase. These reagents were used to develop a sensitive competitive enzyme immunoassay performed in microtitration plates, and the lowest limit of quantification was 150 pg/ml, which is about 10 times more sensitive than previously published techniques. The plasma assay was performed, after a simple methanol extraction, with a minimum of 30 microl of plasma. This assay showed good precision and efficiency, since the rates of recovery from human plasma and cell extracts spiked with ATV ranged form 93 to 113%, with coefficients of variation of less than 10%. ATV concentrations were measured in peripheral blood mononuclear cells incubated with various ATV concentrations and in CEM cells in the absence or presence of antiretroviral drugs and drug transporter inhibitors. The results indicated a dose-dependent uptake (intracellular concentration/extracellular concentration ratio range, 0.04 to 19). A significant increase in the accumulation of ATV was noticed in the presence of P-glycoprotein and MRP1 inhibitors (dipyridamole, inter alia). Interestingly, efavirenz significantly increased the baseline accumulation of ATV, whereas nevirapine induced a marked reduction. This new enzyme immunoassay for measuring plasma and intracellular ATV levels was fully validated and provides an inexpensive and useful tool for routine therapeutic drug monitoring. Moreover, in vitro results suggested the implication of drug transporters and interactions with other antiviral drugs that should be further explored in human immunodeficiency virus infected patients. PMID- 17116662 TI - Novel Ambler class A beta-lactamase LAP-1 and its association with the plasmid mediated quinolone resistance determinant QnrS1. AB - The plasmid-mediated quinolone resistance determinant QnrS1 was identified in non clonally related Enterobacter cloacae isolates in association with a transferable narrow-spectrum beta-lactam resistance marker. Cloning experiments allowed the identification of a novel Ambler class A beta-lactamase, named LAP-1. It shares 62 and 61% amino acid identity with the most closely related beta-lactamases, TEM 1 and SHV-1, respectively. It has a narrow-spectrum hydrolysis of beta-lactams and is strongly inhibited by clavulanic acid and sulbactam and, to a lesser extent, by tazobactam. Association of the blaLAP-1 gene with the qnrS1 gene was identified in E. cloacae isolates from France and Vietnam. These genes were plasmid located and associated with similar insertion sequences but were not associated with sul1-type class 1 integrons, as opposed to the qnrA genes. PMID- 17116663 TI - Development and characterization of a novel single-cycle recombinant-virus assay to determine human immunodeficiency virus type 1 coreceptor tropism. AB - Most human immunodeficiency virus type 1 (HIV-1) strains require either the CXCR4 or CCR5 chemokine receptor to efficiently enter cells. Blocking viral binding to these coreceptors is an attractive therapeutic target. Currently, several coreceptor antagonists are being evaluated in clinical trials that require characterization of coreceptor tropism for enrollment. In this report, we describe the development of an automated and accurate procedure for determining HIV-1 coreceptor tropism (Trofile) and its validation for routine laboratory testing. HIV-1 pseudoviruses are generated using full-length env genes derived from patient virus populations. Coreceptor tropism is determined by measuring the abilities of these pseudovirus populations to efficiently infect CD4+/U87 cells expressing either the CXCR4 or CCR5 coreceptor. Viruses exclusively and efficiently infecting CXCR4+/CD4+/U87 cells are designated X4-tropic. Conversely, viruses exclusively and efficiently infecting CCR5+/CD4+/U87 cells are designated R5-tropic. Viruses capable of infecting both CXCR4+/CD4+/U87 and CCR5+/CD4+/U87 cells are designated dual/mixed-tropic. Assay accuracy and reproducibility were established by evaluating the tropisms of well-characterized viruses and the variability among replicate results from samples tested repeatedly. The viral subtype, hepatitis B virus or hepatitis C virus coinfection, and the plasma viral load did not affect assay performance. Minority subpopulations with alternate tropisms were reliably detected when present at 5 to 10%. The plasma viral load above which samples can be amplified efficiently in the Trofile assay is 1,000 copies per ml of plasma. Trofile has been automated for high-throughput use; it can be used to identify patients most likely to benefit from treatment regimens that include a coreceptor inhibitor and to monitor patients on treatment for the emergence of resistant virus populations that switch coreceptor tropism. PMID- 17116664 TI - Treatment and outcomes of infections by methicillin-resistant Staphylococcus aureus at an ambulatory clinic. AB - Community-acquired methicillin-resistant Staphylococcus aureus (MRSA) skin and soft tissue infections (SSTI) have become increasingly common. This study's objectives were to describe the clinical spectrum of MRSA in a community health center and to determine whether the use of specific antimicrobials correlated with increased probability of clinical resolution of SSTI. A retrospective chart review of 399 sequential cases of culture-confirmed S. aureus SSTI, including 227 cases of MRSA SSTI, among outpatients at Fenway Community Health (Boston, MA) from 1998 to 2005 was done. The proportion of S. aureus SSTI due to MRSA increased significantly from 1998 to 2005 (P<0.0001). Resistance to clindamycin was common (48.2% of isolates). At the beginning of the study period, most patients with MRSA SSTI empirically treated with antibiotics received a beta lactam, whereas by 2005, 76% received trimethoprim-sulfamethoxazole (TMP-SMX) (P<0.0001). Initially, few MRSA isolates were sensitive to the empirical antibiotic, but 77% were susceptible by 2005 (P<0.0001). A significantly higher percentage of patients with MRSA isolates had clinical resolution on the empirical antibiotic by 2005 (P=0.037). Use of an empirical antibiotic to which the clinical isolate was sensitive was associated with increased odds of clinical resolution on empirical therapy (odds ratio=5.91), controlling for incision and drainage and HIV status. MRSA now accounts for the majority of SSTI due to S. aureus at Fenway, and improved rates of clinical resolution on empirical antibiotic therapy have paralleled increasing use of empirical TMP-SMX for these infections. TMP-SMX appears to be an appropriate empirical antibiotic for suspected MRSA SSTI, especially where clindamycin resistance is common. PMID- 17116665 TI - Posaconazole enhances the activity of amphotericin B against Aspergillus hyphae in vitro. AB - The MICs and fractional inhibitory concentrations of posaconazole (POS) and voriconazole (VRZ), alone and in combination with amphotericin B (AMB), for the conidia and hyphae of 100 Aspergillus isolates were evaluated. POS-AMB had more synergistic activity against hyphae (75% of isolates) than VRZ-AMB (37%) and significantly more synergistic activity against hyphae than against conidia (12%). PMID- 17116666 TI - Multistep resistance selection and postantibiotic-effect studies of the antipneumococcal activity of LBM415 compared to other agents. AB - LBM415 is a peptide deformylase inhibitor active against gram-positive bacterial species and some gram-negative species. In multiselection studies, LBM415 had low MICs against all Streptococcus pneumoniae strains tested, regardless of their genotype, and selected resistant clones after 14 to 50 days. MIC increases correlated with changes mostly in the 70GXGXAAXQ77 motif in peptide deformylase. The postantibiotic effect of LBM415 ranged from 0.3 to 1.4 h. PMID- 17116667 TI - In vitro and in vivo activities of Chios mastic gum extracts and constituents against Helicobacter pylori. AB - The extracts and pure major constituents of Chios mastic gum (resin of Pistacia lentiscus var. chia) were tested for their activities against Helicobacter pylori. A total mastic extract without polymer (TMEWP) was prepared after removal of the contained insoluble polymer in order to ameliorate solubility and enhance in vivo activity. Administration of TMEWP to H. pylori SS1-infected mice over the period of 3 months with an average dose of 0.75 mg/day led to an approximately 30 fold reduction in the H. pylori colonization (1.5 log CFU/g of tissue). However, no attenuation in the H. pylori-associated chronic inflammatory infiltration and the activity of chronic gastritis was observed. To further characterize potential active mastic constituents, the TMEWP was separated into an acidic and a neutral fraction. Both were extensively characterized by nuclear magnetic resonance and mass spectroscopy to elucidate the structure of the components contained within each fraction. After chromatographic separation, the acid fraction gave the major triterpenic acids, while the neutral fraction gave several triterpenic alcohols and aldehydes. Mastic extracts and isolated pure triterpenic acids were tested for in vitro activity against a panel of 11 H. pylori clinical strains. The acid fraction was found to be the most active extract (minimum bactericidal concentration [MBC], 0.139 mg/ml), and the most active pure compound was isomasticadienolic acid (MBC, 0.202 mg/ml [0.443 mM]). Our results show that administration of TMEWP may be effective in reducing H. pylori colonization and that the major triterpenic acids in the acid extract may be responsible for such an activity. PMID- 17116668 TI - Fitness costs of fluoroquinolone resistance in Streptococcus pneumoniae. AB - The fitness cost of the genes responsible for resistance to fluoroquinolones in clinical isolates of Streptococcus pneumoniae were estimated in vitro in a common genetic background. Naturally occurring parC, parE, and gyrA loci containing mutations in the quinolone-resistance-determining regions were introduced by transformation into S. pneumoniae strain R6 individually and in combinations. The fitness of these transformants was estimated by pairwise competition experiments with a common R6 strain. On average, single par and gyr mutants responsible for low-level MIC resistance (first-step resistance) impose a fitness burden of approximately 8%. Some of these mutants engender no measurable cost, while one, a parE mutant, reduces the fitness of these bacteria by more than 40%. Most interestingly, the addition of the second par or gyr mutations required for clinically significant, high-MIC fluoroquinolone resistance does not increase the fitness burden imposed by these single genes and can even reduce it. We discuss the implications of these results for the epidemiology of fluoroquinolone resistance and the evolution of acquired resistance in treated patients. PMID- 17116669 TI - In vitro antimalarial activity of azithromycin, artesunate, and quinine in combination and correlation with clinical outcome. AB - Azithromycin when used in combination with faster-acting antimalarials has proven efficacious in treating Plasmodium falciparum malaria in phase 2 clinical trials. The aim of this study was to establish optimal combination ratios for azithromycin in combination with either dihydroartemisinin or quinine, to determine the clinical correlates of in vitro drug sensitivity for these compounds, and to assess the cross-sensitivity patterns. Seventy-three fresh P. falciparum isolates originating from patients from the western border regions of Thailand were successfully tested for their drug susceptibility in a histidine rich protein 2 (HRP2) assay. With overall mean fractional inhibitory concentrations of 0.84 (95% confidence interval [CI]=0.77 to 1.08) and 0.78 (95% CI=0.72 to 0.98), the interactions between azithromycin and dihydroartemisinin, as well as quinine, were classified as additive, with a tendency toward synergism. The strongest tendency toward synergy was seen with a combination ratio of 1:547 for the combination with dihydroartemisinin and 1:44 with quinine. The geometric mean 50% inhibitory concentration (IC50) of azithromycin was 2,570.3 (95% CI=2,175.58 to 3,036.58) ng/ml. The IC50s for mefloquine, quinine, and chloroquine were 11.42, 64.4, and 54.4 ng/ml, respectively, suggesting a relatively high level of background resistance in this patient population. Distinct correlations (R=0.53; P=0.001) between quinine in vitro results and parasite clearance may indicate a compromised sensitivity to this drug. The correlation with dihydroartemisinin data was weaker (R=0.34; P=0.038), and no such correlation was observed for azithromycin. Our in vitro data confirm that azithromycin in combination with artemisinin derivatives or quinine exerts additive to synergistic interactions, shows no cross-sensitivity with traditional antimalarials, and has substantial antimalarial activity on its own. PMID- 17116670 TI - Steady-state pharmacokinetics of micafungin and voriconazole after separate and concomitant dosing in healthy adults. AB - We assessed the pharmacokinetics and interactions of steady-state micafungin (Mycamine) or placebo with steady-state voriconazole in 35 volunteers. The 90% confidence intervals around the least-squares mean ratios for micafungin pharmacokinetic parameters and placebo-corrected voriconazole pharmacokinetic parameters were within the 80%-to-125% limits, indicating an absence of drug interaction. PMID- 17116671 TI - RNAIII-inhibiting-peptide-loaded polymethylmethacrylate prevents in vivo Staphylococcus aureus biofilm formation. AB - Staphylococci, common orthopedic pathogens, form antibiotic-resistant biofilms. Polymethylmethacrylate (PMMA) beads loaded with the quorum-sensing inhibitor RNAIII-inhibiting peptide (RIP) were implanted in rats and shown to prevent methicillin-resistant Staphylococcus aureus infection. RIP release was bimodal, typical of previously-tested antibiotics. These results suggest that RIP-PMMA warrants further evaluation for management of orthopedic infections caused by staphylococci. PMID- 17116672 TI - Development of a new methodology for screening of human immunodeficiency virus type 1 microbicides based on real-time PCR quantification. AB - Potential topical retrovirucides or vaginal microbicides against human immunodeficiency virus type 1 (HIV-1) include nonnucleoside reverse transcriptase inhibitors (NNRTIs). To be successful, such agents have to be highly active against cell-free virions. In the present study, we developed a new real-time PCR based assay to measure the natural endogenous reverse transcription (NERT) activity directly on intact HIV-1 particles in the presence of reverse transcriptase (RT) inhibitors. We further evaluated the permeability to nevirapine (NVP) and efavirenz (EFV) and their retention within nascent viral particles. We also demonstrated the NVP and EFV inhibitory effects on NERT activity and the impact of resistance mutations measured directly by this new strategy. Furthermore, the results showed a clear correlation between NERT activity and classical infectivity assays. The 50% inhibitory concentrations (IC50s) of NVP and EFV were demonstrated to be up to 100-fold higher for cell free than for cell-associated virions, suggesting that cell-free virions are less permeable to these drugs. Our results suggest that NVP and EFV penetrate both the envelope and the capsid of HIV-1 particles and readily inactivate cell-free virions. However, the characteristics of these NNRTIs, such as lower permeability and lower retention during washing procedures, in cell-free virions reduce their efficacies as microbicides. Here, we demonstrate the usefulness of the NERT real time PCR as an assay for screening novel antiretroviral compounds with unique mechanisms of action. PMID- 17116673 TI - Isolation and characterization of human immunodeficiency virus type 1 resistant to the small-molecule CCR5 antagonist TAK-652. AB - TAK-652, a novel small-molecule chemokine receptor antagonist, is a highly potent and selective inhibitor of CCR5-using (R5) human immunodeficiency virus type 1 (HIV-1) replication in vitro. Since TAK-652 is orally bioavailable and has favorable pharmacokinetic profiles in humans, it is considered a promising candidate for an entry inhibitor of HIV-1. To investigate the resistance to TAK 652, peripheral blood mononuclear cells were infected with the R5 HIV-1 primary isolate KK and passaged in the presence of escalating concentrations of the compound for more than 1 year. After 67 weeks of cultivation, the escape virus emerged even in the presence of a high concentration of TAK-652. This virus displayed more than 200,000-fold resistance to TAK-652 compared with the wild type. The escape virus appeared to have cross-resistance to the structurally related compound TAK-779 but retained full susceptibility to TAK-220, which is from a different class of CCR5 antagonists. Furthermore, the escape virus was unable to use CXCR4 as a coreceptor. Analysis for Env amino acid sequences of escape viruses at certain points of passage revealed that amino acid changes accumulated with an increasing number of passages. Several amino acid changes not only in the V3 region but also in other Env regions seemed to be required for R5 HIV-1 to acquire complete resistance to TAK-652. PMID- 17116674 TI - Natural polymorphisms in the human immunodeficiency virus type 2 protease can accelerate time to development of resistance to protease inhibitors. AB - Human immunodeficiency virus type 2 (HIV-2) contains numerous natural polymorphisms in its protease (PR) gene that are implicated in drug resistance in the case of HIV-1. This study evaluated emergent PR resistance in HIV-2. Three HIV-2 isolates were selected for resistance to amprenavir (APV), nelfinavir (NFV), indinavir (IDV), and tipranavir (TPV) in cell culture. Genotypic analysis determined the time to the appearance of protease inhibitor (PI)-associated mutations compared to HIV-1. Phenotypic drug susceptibility assays were used to determine the levels of drug resistance. Within 10 to 15 weeks of serial passage, three major mutations--I54M, I82F, and L90M--arose in HIV-2 viral cultures exposed to APV, NFV, and IDV, whereas I82L was selected with TPV. After 25 weeks, other cultures had developed I50V and I84V mutations. In contrast, no major PI mutations were selected in HIV-1 over this period except for D30N in the context of NFV selective pressure. The baseline phenotypes of wild-type HIV-2 isolates were in the range observed for HIV-1, except for APV and NFV for which a lower degree of sensitivity was seen. The acquisition of the I54M, I84V, L90M, and L99F mutations resulted in multi-PI-resistant viruses, conferring 10-fold to more than 100-fold resistance. Of note, we observed a 62A/99F mutational motif that conferred high-level resistance to PIs, as well as novel secondary mutations, including 6F, 12A, and 21K. Thus, natural polymorphisms in HIV-2 may facilitate the selection of PI resistance. The increasing incidence of such polymorphisms in drug-naive HIV-1- and HIV-2-infected persons is of concern. PMID- 17116675 TI - Dual targeting of GyrB and ParE by a novel aminobenzimidazole class of antibacterial compounds. AB - A structure-guided drug design approach was used to optimize a novel series of aminobenzimidazoles that inhibit the essential ATPase activities of bacterial DNA gyrase and topoisomerase IV and that show potent activities against a variety of bacterial pathogens. Two such compounds, VRT-125853 and VRT-752586, were characterized for their target specificities and preferences in bacteria. In metabolite incorporation assays, VRT-125853 inhibited both DNA and RNA synthesis but had little effect on protein synthesis. Both compounds inhibited the maintenance of negative supercoils in plasmid DNA in Escherichia coli at the MIC. Sequencing of DNA corresponding to the GyrB and ParE ATP-binding regions in VRT 125853- and VRT-752586-resistant mutants revealed that their primary target in Staphylococcus aureus and Haemophilus influenzae was GyrB, whereas in Streptococcus pneumoniae it was ParE. In Enterococcus faecalis, the primary target of VRT-125853 was ParE, whereas for VRT-752586 it was GyrB. DNA transformation experiments with H. influenzae and S. aureus proved that the mutations observed in gyrB resulted in decreased susceptibilities to both compounds. Novobiocin resistance-conferring mutations in S. aureus, H. influenzae, and S. pneumoniae were found in gyrB, and these mutants showed little or no cross-resistance to VRT-125853 or VRT-752586 and vice versa. Furthermore, gyrB and parE double mutations increased the MICs of VRT-125853 and VRT-752586 significantly, providing evidence of dual targeting. Spontaneous frequencies of resistance to VRT-752586 were below detectable levels (<5.2x10(-10)) for wild type E. faecalis but were significantly elevated for strains containing single and double target-based mutations, demonstrating that dual targeting confers low levels of resistance emergence and the maintenance of susceptibility in vitro. PMID- 17116676 TI - High-throughput Plasmodium falciparum growth assay for malaria drug discovery. AB - New therapeutic agents for the treatment of malaria, the world's most deadly parasitic disease, are urgently needed. Malaria afflicts 300 to 500 million people and results in 1 to 2 million deaths annually, and more than 85% of all malaria-related mortality involves young children and pregnant women in sub Saharan Africa. The emergence of multidrug-resistant parasites, especially in Plasmodium falciparum, has eroded the efficacy of almost all currently available therapeutic agents. The discovery of new drugs, including drugs with novel cellular targets, could be accelerated with a whole-organism high-throughput screen (HTS) of structurally diverse small-molecule libraries. The standard whole organism screen is based on incorporation of [3H]hypoxanthine and has liabilities, such as limited throughput, high cost, multiple labor-intensive steps, and disposal of radioactive waste. Recently, screens have been reported that do not use radioactive incorporation, but their reporter signal is not robust enough for HTS. We report a P. falciparum growth assay that is technically simple, robust, and compatible with the automation necessary for HTS. The assay monitors DNA content by addition of the fluorescent dye 4',6-diamidino-2 phenylindole (DAPI) as a reporter of blood-stage parasite growth. This DAPI P. falciparum growth assay was used to measure the 50% inhibitory concentrations (IC50s) of a diverse set of known antimalarials. The resultant IC50s compared favorably with those obtained in the [3H]hypoxanthine incorporation assay. Over 79,000 small molecules have been tested for antiplasmodial activity using the DAPI P. falciparum growth assay, and 181 small molecules were identified as highly active against multidrug-resistant parasites. PMID- 17116677 TI - A novel nonnucleoside analogue that inhibits human immunodeficiency virus type 1 isolates resistant to current nonnucleoside reverse transcriptase inhibitors. AB - Nonnucleoside reverse transcriptase (RT) inhibitors (NNRTIs) are important components of current combination therapies for human immunodeficiency virus type 1 (HIV-1) infection. However, their low genetic barriers against resistance development, cross-resistance, and serious side effects can compromise the benefits of the two current drugs in this class (efavirenz and nevirapine). In this study, we report a novel and potent NNRTI, VRX-480773, that inhibits viruses from efavirenz-resistant molecular clones and most NNRTI-resistant clinical HIV-1 isolates tested. In vitro mutation selection experiments revealed that longer times were required for viruses to develop resistance to VRX-480773 than to efavirenz. RT mutations selected by VRX-480773 after 3 months of cell culture in the presence of 1 nM VRX-480773 carried the Y181C mutation, resulting in a less than-twofold increase in resistance to the compound. A virus containing the double mutation V106I-Y181C emerged after 4 months, causing a sixfold increase in resistance. Viruses containing additional mutations of D123G, F227L, and T369I emerged when the cultures were incubated with increasing concentrations of VRX 480773. Most of the resistant viruses selected by VRX-480773 are susceptible to efavirenz. Oral administration of VRX-480773 to dogs resulted in plasma concentrations that were significantly higher than those required for the inhibition of wild-type and mutant viruses. These results warrant further clinical development of VRX-480773 for the treatment of HIV infection in both NNRTI-naive and -experienced patients. PMID- 17116678 TI - Leishmania donovani polyamine biosynthetic enzyme overproducers as tools to investigate the mode of action of cytotoxic polyamine analogs. AB - A number of anticancer and antiparasitic drugs are postulated to target the polyamine biosynthetic pathway and polyamine function, but the exact mode of action of these compounds is still being elucidated. To establish whether polyamine analogs specifically target enzymes of the polyamine pathway, a model was developed using strains of the protozoan parasite Leishmania donovani that overproduce each of the polyamine biosynthetic enzymes. Promastigotes overexpressing episomal constructs encoding ornithine decarboxylase (ODC), S adenosylmethionine decarboxylase (ADOMETDC), or spermidine synthase (SPDSYN) revealed robust overproduction of the corresponding polyamine biosynthetic enzyme. Polyamine pools, however, were either unchanged or only marginally affected, implying that regulatory mechanisms must exist. The ODC, ADOMETDC, and SPDSYN overproducer strains exhibited a high level of resistance to difluoromethylornithine, 5'-{[(Z)-4-amino-2-butenyl]methylamino}-5' deoxyadenosine, and n-butylamine, respectively, confirming previous observations that these agents specifically target polyamine enzymes. Conversely, augmented levels of polyamine biosynthetic enzymes did not affect the sensitivity of L. donovani promastigotes to pentamidine, berenil, and mitoguazone, drugs that were postulated to target the polyamine pathway, implying alternative and/or additional targets for these agents. The sensitivities of wild-type and overproducing parasites to a variety of polyamine analogs were also tested. The polyamine enzyme-overproducing lines offer a rapid cell-based screen for assessing whether synthetic polyamine analogs exert their mechanism of action predominantly on the polyamine biosynthetic pathway in L. donovani. Furthermore, the drug resistance engendered by the amplification of target genes and the overproduction of the encoded protein offers a general strategy for evaluating and developing therapeutic agents that target specific proteins in Leishmania. PMID- 17116679 TI - The relationship between quinolone exposures and resistance amplification is characterized by an inverted U: a new paradigm for optimizing pharmacodynamics to counterselect resistance. AB - We determined the relationship between garenoxacin exposure and quinolone resistant subpopulations for three bacterial isolates in an in vitro hollow-fiber infection model. An "inverted-U" relationship was identified wherein resistant subpopulations rose initially and then declined with increasing exposure, until reaching a threshold that prevented resistance amplifications. Different targets for the area under the concentration-time curve over 24 h/MIC ratio were required for different bacteria. PMID- 17116680 TI - Comparative DNA analysis of two vanA plasmids from Enterococcus faecium strains isolated from poultry and a poultry farmer in Norway. AB - The DNA sequences of two plasmids carrying vanA, pVEF1 (39,626 bp) and pVEF2 (39,714 bp), were determined. Forty-three shared coding sequences were identified, and the only nucleotide difference was an 88-bp indel. A postsegregational killing system was identified. This system possibly explains the persistence of the vanA gene cluster in Norwegian poultry farms. PMID- 17116681 TI - Diversity of ampicillin resistance genes and antimicrobial susceptibility patterns in Haemophilus influenzae strains isolated in Korea. AB - By Etest determination of the susceptibilities of 229 Haemophilus influenzae strains isolated in Korea to 10 antibiotics, the isolates were found to be antibiotic nonsusceptible in the following order: ampicillin (58.1%), trimethoprim-sulfamethoxazole (52%), cefaclor (41.1%), clarithromycin (25.8%), chloramphenicol (14.0%), amoxicillin-clavulanic acid (13.5%), meropenem (11.7%), cefixime (10.9%), cefuroxime (9.2%), and levofloxacin (1.3%). The prevalences of each resistance class were 23.6% for beta-lactamase-negative ampicillin susceptible (BLNAS) strains; 37.6% for strains with the TEM-1 type beta-lactamase gene; 1.3% for strains with the ROB-1 type beta-lactamase gene; 29.3% for the beta-lactamase-negative ampicillin-resistant (BLNAR) strains with a mutation in the ftsI gene, which encodes PBP 3; and 8.3% for beta-lactamase-positive amoxicillin-clavulanate-resistant (BLPACR) strains, which showed both resistance mechanisms (i.e., a beta-lactamase gene and a mutation in the ftsI gene). The MIC50s of all beta-lactams, including cephem and meropenem agents, for the BLNAR strains were two to three times higher than those for the BLNAS strains. This study confirms that the prevalence of BLNAR and BLPACR strains is relatively high and for the first time confirms the presence of H. influenzae strains carrying blaROB-1 in Korea. Even though mutations in another gene(s) might be involved in beta-lactam resistance, these results suggest that mutations in the ftsI gene are important for the development of resistance to beta-lactams in H. influenzae strains in Korea. PMID- 17116682 TI - Caenorhabditis elegans as a model to determine fitness of antibiotic-resistant Salmonella enterica serovar typhimurium. AB - We used the ability of Salmonella enterica serovar Typhimurium to colonize the gut of Caenorhabditis elegans to measure the fitness costs imposed by antibiotic resistance mutations. The fitness costs determined in the nematode were similar to those measured in mice, validating its use as a simple host model to evaluate bacterial fitness. PMID- 17116683 TI - Efficacy of delayed treatment with ST-246 given orally against systemic orthopoxvirus infections in mice. AB - ST-246 was evaluated for activity against cowpox virus (CV), vaccinia virus (VV), and ectromelia virus (ECTV) and had an in vitro 50% effective concentration (EC50) of 0.48 microM against CV, 0.05 microM against VV, and 0.07 microM against ECTV. The selectivity indices were >208 and >2,000 for CV and VV, respectively. The in vitro antiviral activity of ST-246 was significantly greater than that of cidofovir, which had an EC50 of 41.1 microM against CV and 29.2 microM against VV, with selectivity indices of >7 and >10, respectively. ST-246 administered once daily by oral gavage to mice infected intranasally with CV beginning 4 h or delayed until 72 h postinoculation was highly effective when given for a 14-day duration using 100, 30, or 10 mg/kg of body weight. When 100 mg/kg of ST-246 was administered to VV-infected mice, a duration of 5 days was sufficient to significantly reduce mortality even when treatment was delayed 24 h postinoculation. Viral replication in liver, spleen, and kidney, but not lung, of CV- or VV-infected mice was reduced by ST-246 compared to levels for vehicle treated mice. When 100 mg/kg of ST-246 was given once daily to mice infected by the intranasal route with ECTV, treatment for 10 days prevented mortality even when treatment was delayed up to 72 h after viral inoculation. Viral replication in target organs of ECTV-infected mice was also reduced. PMID- 17116684 TI - Tolerance to the glycopeptides vancomycin and teicoplanin in coagulase-negative staphylococci. AB - Tolerance to vancomycin and teicoplanin in 90 clinical isolates of coagulase negative staphylococci (CoNS) was investigated by time-kill curve methodology. Only six strains, belonging to the Staphylococcus lugdunensis species, exhibited tolerance. The seven other S. lugdunensis strains tested displayed weak susceptibility to the bactericidal activity of glycopeptides compared to the other CoNS. These phenomena are of concern, since S. lugdunensis is recognized as one of the most pathogenic CoNS. PMID- 17116685 TI - Estimating true antimalarial efficacy by heteroduplex tracking assay in patients with complex Plasmodium falciparum infections. AB - Heteroduplex tracking assays (HTAs) of Plasmodium falciparum merozoite surface protein 1 block-2 were used to assess complexity of infection and treatment efficacy in a trial of three antimalarial treatments in 141 Malawian pregnant women. An elevated complexity of infection (COI) was associated with anemia, parasite burden, and human immunodeficiency virus infection but was not associated with age or gravidity. Comparisons of HTA patterns before and after treatment allowed the classification of 20 of 30 (66%) recurrent episodes as either definite treatment failures or reinfections. An elevated COI was strongly associated with treatment failure (P=0.003). An algorithm was developed to assign a probability of failure for the 10 indeterminate participants, some of whose infections shared a single variant of high prevalence (>10%). By summing these probabilities, treatment efficacy was estimated. PMID- 17116686 TI - Efficacy of bacteriophage therapy against gut-derived sepsis caused by Pseudomonas aeruginosa in mice. AB - We evaluated the efficacy of bacteriophage (phage) therapy by using a murine model of gut-derived sepsis caused by Pseudomonas aeruginosa that closely resembles the clinical pathophysiology of septicemia in humans. Oral administration of a newly isolated lytic phage strain (KPP10) significantly protected mice against mortality (survival rates, 66.7% for the phage-treated group versus 0% for the saline-treated control group; P<0.01). Mice treated with phage also had lower numbers of viable P. aeruginosa cells in their blood, liver, and spleen. The levels of inflammatory cytokines (tumor necrosis factor alpha TNF alpha, interleukin-1beta [IL-1beta], and IL-6) in blood and liver were significantly lower in phage-treated mice than in phage-untreated mice. The number of viable P. aeruginosa cells in fecal matter in the gastrointestinal tract was significantly lower in phage-treated mice than in the saline-treated control mice. We also studied the efficacy of phage treatment for intraperitoneal infection caused by P. aeruginosa and found that phage treatment significantly improved the survival of mice, but only under limited experimental conditions. In conclusion, our findings suggest that oral administration of phage may be effective against gut-derived sepsis caused by P. aeruginosa. PMID- 17116687 TI - Abr and Bcr, two homologous Rac GTPase-activating proteins, control multiple cellular functions of murine macrophages. AB - Small GTPases of the Rho family are key regulators of phagocytic leukocyte function. Abr and Bcr are homologous, multidomain proteins. Their C-terminal domain has GTPase-activating protein (GAP) activity that, in vitro, is specific for Rac and Cdc42. To address the in vivo relevance of these entire proteins, of which little is known, the current study examined the effect of the genetic ablation of Abr and Bcr in murine macrophages. The concomitant loss of Abr and Bcr induced multiple alterations of macrophage cellular behavior known to be under the control of Rac. Macrophages lacking both Abr and Bcr exhibited an atypical, elongated morphology that was reproduced by the ectopic expression of GAP domain mutant Abr and Bcr in a macrophage cell line and of constitutively active Rac in primary macrophages. A robust increase in colony-stimulating factor 1 (CSF-1)-directed motility was observed in macrophages deficient for both proteins and, in response to CSF-1 stimulation, Abr and Bcr transiently translocated to the plasma membrane. Phagocytosis of opsonized particles was also increased in macrophages lacking both proteins and correlated with sustained Rac activation. Bcr and Abr GAP mutant proteins localized around phagosomes and induced distinct phagocytic cup formation. These results identify Abr and Bcr as the only GAPs to date that specifically negatively regulate Rac function in vivo in primary macrophages. PMID- 17116688 TI - A two-step, PU.1-dependent mechanism for developmentally regulated chromatin remodeling and transcription of the c-fms gene. AB - Hematopoietic stem cells and multipotent progenitors exhibit low-level transcription and partial chromatin reorganization of myeloid cell-specific genes including the c-fms (csf1R) locus. Expression of the c-fms gene is dependent on the Ets family transcription factor PU.1 and is upregulated during myeloid differentiation, enabling committed macrophage precursors to respond to colony stimulating factor 1. To analyze molecular mechanisms underlying the transcriptional priming and developmental upregulation of the c-fms gene, we have utilized myeloid progenitors lacking the transcription factor PU.1. PU.1 can bind to sites in both the c-fms promoter and the c-fms intronic regulatory element (FIRE enhancer). Unlike wild-type progenitors, the PU.1(-/-) cells are unable to express c-fms or initiate macrophage differentiation. When PU.1 was reexpressed in mutant progenitors, the chromatin structure of the c-fms promoter was rapidly reorganized. In contrast, assembly of transcription factors at FIRE, acquisition of active histone marks, and high levels of c-fms transcription occurred with significantly slower kinetics. We demonstrate that the reason for this differential activation was that PU.1 was required to promote induction and binding of a secondary transcription factor, Egr-2, which is important for FIRE enhancer activity. These data suggest that the c-fms promoter is maintained in a primed state by PU.1 in progenitor cells and that at FIRE PU.1 functions with another transcription factor to direct full activation of the c-fms locus in differentiated myeloid cells. The two-step mechanism of developmental gene activation that we describe here may be utilized to regulate gene activity in a variety of developmental pathways. PMID- 17116689 TI - Cancer-associated mutations in the MDM2 zinc finger domain disrupt ribosomal protein interaction and attenuate MDM2-induced p53 degradation. AB - The p53-inhibitory function of the oncoprotein MDM2 is regulated by a number of MDM2-binding proteins, including ARF and ribosomal proteins L5, L11, and L23, which bind the central acidic domain of MDM2 and inhibit its E3 ubiquitin ligase activity. Various human cancer-associated MDM2 alterations targeting the central acidic domain have been reported, yet the functional significance of these mutations in tumor development has remained unclear. Here, we show that cancer associated missense mutations targeting MDM2's central zinc finger disrupt the interaction of MDM2 with L5 and L11. We found that the zinc finger mutant MDM2 is impaired in undergoing nuclear export and proteasomal degradation as well as in promoting p53 degradation, yet retains the function of suppressing p53 transcriptional activity. Unlike the wild-type MDM2, whose p53-suppressive activity can be inhibited by L11, the MDM2 zinc finger mutant escapes L11 inhibition. Hence, the MDM2 central zinc finger plays a critical role in mediating MDM2's interaction with ribosomal proteins and its ability to degrade p53, and these roles are disrupted by human cancer-associated MDM2 mutations. PMID- 17116690 TI - Intramolecular regulation of phospholipase C-gamma1 by its C-terminal Src homology 2 domain. AB - Phosphoinositide-specific phospholipase C-gamma1 (PLC-gamma1) is a key enzyme that governs cellular functions such as gene transcription, secretion, proliferation, motility, and development. Here, we show that PLC-gamma1 is regulated via a novel autoinhibitory mechanism involving its carboxy-terminal Src homology (SH2C) domain. Mutation of the SH2C domain tyrosine binding site led to constitutive PLC-gamma1 activation. The amino-terminal split pleckstrin homology (sPHN) domain was found to regulate the accessibility of the SH2C domain. PLC gamma1 constructs with mutations in tyrosine 509 and phenylalanine 510 in the sPHN domain no longer required an intact amino-terminal Src homology (SH2N) domain or phosphorylation of tyrosine 775 or 783 for activation. These data are consistent with a model in which the SH2C domain is blocked by an intramolecular interaction(s) that is released upon cellular activation by occupancy of the SH2N domain. PMID- 17116691 TI - STAMP, a novel predicted factor assisting TIF2 actions in glucocorticoid receptor mediated induction and repression. AB - The coactivator TIF2 was predicted to interact with an unknown factor to modify both the relative inhibition in glucocorticoid receptor (GR)-mediated gene repression and several parameters of agonists and antisteroids in GR-regulated induction. Here, we describe the isolation and characterization of the predicted factor as a new 1,277-amino-acid endogenous protein (STAMP). STAMP associates with coactivators (TIF2 and SRC-1) and is selective for a subset of the steroid/nuclear receptors including GRs. Transfected STAMP increases the effects of TIF2 in GR-mediated repression and induction. Conversely, the levels of both induction and repression of endogenous genes are reduced when STAMP small interfering RNAs are used to lower the level of endogenous STAMP. Endogenous STAMP colocalizes with GR in intact cells and is recruited to the promoters of endogenous GR-induced and -repressed genes. We suggest that STAMP is an important new, downstream component of GR action in both gene activation and gene repression. PMID- 17116692 TI - Novel role for Cdc14 sequestration: Cdc14 dephosphorylates factors that promote DNA replication. AB - The phosphatase Cdc14 is required for mitotic exit in budding yeast. Cdc14 promotes Cdk1 inactivation by targeting proteins that, when dephosphorylated, trigger degradation of mitotic cyclins and accumulation of the Cdk1 inhibitor, Sic1. Cdc14 is sequestered in the nucleolus during most of the cell cycle but is released into the nucleus and cytoplasm during anaphase. When Cdc14 is not properly sequestered in the nucleolus, expression of the S-phase cyclin Clb5 is required for viability, suggesting that the antagonizing activity of Clb5 dependent Cdk1 specifically is necessary when Cdc14 is delocalized. We show that delocalization of Cdc14 combined with loss of Clb5 causes defects in DNA replication. When Cdc14 is not sequestered, it efficiently dephosphorylates a subset of Cdk1 substrates including the replication factors, Sld2 and Dpb2. Mutations causing Cdc14 mislocalization interact genetically with mutations affecting the function of DNA polymerase epsilon and the S-phase checkpoint protein Mec1. Our findings suggest that Cdc14 is retained in the nucleolus to support a favorable kinase/phosphatase balance while cells are replicating their DNA, in addition to the established role of Cdc14 sequestration in coordinating nuclear segregation with mitotic exit. PMID- 17116693 TI - Helix-loop-helix protein p8, a transcriptional regulator required for cardiomyocyte hypertrophy and cardiac fibroblast matrix metalloprotease induction. AB - Cardiomyocyte hypertrophy and extracellular matrix remodeling, primarily mediated by inflammatory cytokine-stimulated cardiac fibroblasts, are critical cellular events in cardiac pathology. The molecular components governing these processes remain nebulous, and few genes have been linked to both hypertrophy and matrix remodeling. Here we show that p8, a small stress-inducible basic helix-loop-helix protein, is required for endothelin- and alpha-adrenergic agonist-induced cardiomyocyte hypertrophy and for tumor necrosis factor-stimulated induction, in cardiac fibroblasts, of matrix metalloproteases (MMPs) 9 and 13-MMPs linked to general inflammation and to adverse ventricular remodeling in heart failure. In a stimulus-dependent manner, p8 associates with chromatin containing c-Jun and with the cardiomyocyte atrial natriuretic factor (anf) promoter and the cardiac fibroblast mmp9 and mmp13 promoters, established activator protein 1 effectors. p8 is also induced strongly in the failing human heart by a process reversed upon therapeutic intervention. Our results identify an unexpectedly broad involvement for p8 in key cellular events linked to cardiomyocyte hypertrophy and cardiac fibroblast MMP production, both of which occur in heart failure. PMID- 17116694 TI - Gene trap disruption of the mouse heparan sulfate 6-O-endosulfatase gene, Sulf2. AB - Heparan sulfate (HS) chains are found in the extracellular matrix, covalently linked to core proteins collectively termed heparan sulfate proteoglycans (HSPGs). A wealth of data has demonstrated roles for HSPGs in the regulation of many cell surface signaling pathways that are crucial during development. Variations in the sulfation pattern along the HS chains influence their ability to interact with molecules such as growth factors, chemokines, morphogens, and adhesion molecules. Sulf1 and Sulf2 are members of a class of recently identified genes that encode heparan sulfate 6-O-endosulfatases (Sulf genes). The removal of 6-O-sulfate from HS via SULF activity influences the function of many factors, including Wnt, fibroblast growth factor, hepatocyte growth factor, heparin binding epidermal growth factor, and bone morphogenetic protein. Given their possible developmental roles, we have examined Sulf gene expression during mouse embryogenesis. The two Sulf genes are expressed in a broad range of tissues throughout development with largely nonoverlapping expression patterns. Sulf2 transcripts are expressed in the lung, heart, placenta, and ribs. We generated a mouse line possessing a gene trap disruption of the Sulf2 gene. Mice homozygous for the Sulf2 gene trap allele are viable and fertile and have no major developmental defects on several genetic backgrounds. However, we observed strain specific, nonpenetrant defects affecting viability, lung development, and growth in Sulf2 homozygous animals. These data suggest that Sulf2 may have roles in several tissues but that there is compensation by and/or redundancy with Sulf1. PMID- 17116695 TI - Nuclear export of the transcription factor NirA is a regulatory checkpoint for nitrate induction in Aspergillus nidulans. AB - NirA, the specific transcription factor of the nitrate assimilation pathway of Aspergillus nidulans, accumulates in the nucleus upon induction by nitrate. NirA interacts with the nuclear export factor KapK, which bridges an interaction with a protein of the nucleoporin-like family (NplA). Nitrate induction disrupts the NirA-KapK interaction in vivo, whereas KapK associates with NirA when this protein is exported from the nucleus. A KpaK leptomycin-sensitive mutation leads to inducer-independent NirA nuclear accumulation in the presence of the drug. However, this does not lead to constitutive expression of the genes controlled by NirA. A nirA(c)1 mutation leads to constitutive nuclear localization and activity, remodeling of chromatin, and in vivo binding to a NirA upstream activation sequence. The nirA(c)1 mutation maps in the nuclear export signal (NES) of the NirA protein. The NirA-KapK interaction is nearly abolished in NirA(c)1 and NirA proteins mutated in canonical leucine residues in the NirA NES. The latter do not result in constitutively active NirA protein, which implies that nuclear retention is necessary but not sufficient for NirA activity. The results are consistent with a model in which activation of NirA by nitrate disrupts the interaction of NirA with the NplA/KapK nuclear export complex, thus resulting in nuclear retention, leading to AreA-facilitated DNA binding of the NirA protein and subsequent chromatin remodeling and transcriptional activation. PMID- 17116697 TI - Limited value of plain radiographs in infant torticollis. AB - OBJECTIVE: The purpose of this work was to assess the frequency of clinically relevant findings from plain films of infants evaluated for torticollis. PATIENTS AND METHODS: After institutional review board approval, radiology records were searched for infants 0 to 12 months of age who underwent plain film study for torticollis or "head tilt." Infants evaluated for trauma or Down syndrome were excluded. All of the studies were reviewed, demographic data was recorded, and any additional imaging studies were examined. RESULTS: A total of 502 patients (189 girls and 313 boys) were identified with an average age of 0.37 +/- 0.2 years. Head tilt was to the left in two thirds of patients. Ten patients had abnormal findings reported. Six of these proved normal on subsequent studies (3 suspected occipital-C1 fusions, 2 suspected cervical fusions, and 1 suspected hemivertebra). Four patients had true bony vertebral abnormalities including absent left C7 pedicle, multiple fusion anomalies from C4 to T2, C3 hemivertebra and thoracic spine anomalies, and C4 hypoplasia. This last patient had abnormal kyphosis on physical examination and demonstrated instability with dynamic testing. Twenty-five additional patients with normal plain films underwent spine computed tomography or magnetic resonance imaging; all were normal. CONCLUSIONS: The true-positive yield of plain films in nontraumatic infant torticollis was low (4 of 502). There were more false-positive than true-positive results. A common rationale for imaging is to exclude craniocervical or other unstable abnormalities that might contraindicate physical therapy, seen in only 1 of the 502 cases. Close physical examination could safely eliminate most patients sent for radiography. PMID- 17116696 TI - DNA recombination-initiation plays a role in the extremely biased inheritance of yeast [rho-] mitochondrial DNA that contains the replication origin ori5. AB - Hypersuppressiveness, as observed in Saccharomyces cerevisiae, is an extremely biased inheritance of a small mitochondrial DNA (mtDNA) fragment that contains a replication origin (HS [rho(-)] mtDNA). Our previous studies showed that concatemers (linear head-to-tail multimers) are obligatory intermediates for mtDNA partitioning and are primarily formed by rolling-circle replication mediated by Mhr1, a protein required for homologous mtDNA recombination. In this study, we found that Mhr1 is required for the hypersuppressiveness of HS [ori5] [rho(-)] mtDNA harboring ori5, one of the replication origins of normal ([rho(+)]) mtDNA. In addition, we detected an Ntg1-stimulated double-strand break at the ori5 locus. Purified Ntg1, a base excision repair enzyme, introduced a double-stranded break by itself into HS [ori5] [rho(-)] mtDNA at ori5 isolated from yeast cells. Both hypersuppressiveness and concatemer formation of HS [ori5] [rho(-)] mtDNA are simultaneously suppressed by the ntg1 null mutation. These results support a model in which, like homologous recombination, rolling-circle HS [ori5] [rho(-)] mtDNA replication is initiated by double-stranded breakage in ori5, followed by Mhr1-mediated homologous pairing of the processed nascent DNA ends with circular mtDNA. The hypersuppressiveness of HS [ori5] [rho(-)] mtDNA depends on a replication advantage furnished by the higher density of ori5 sequences and on a segregation advantage furnished by the higher genome copy number on transmitted concatemers. PMID- 17116698 TI - Long-term cognitive and motor deficits after enterovirus 71 brainstem encephalitis in children. AB - OBJECTIVES: Several large outbreaks of enterovirus 71 infections have occurred in Taiwan during the past decade. Brainstem encephalitis was the most common neurologic complication. This study was designed to determine the long-term cognitive and motor outcomes of children with enterovirus 71 brainstem encephalitis. METHODS: We conducted a prospective follow-up study of children who met the case definition for enterovirus 71 brainstem encephalitis. Subjects were stratified into isolated brainstem encephalitis (stage II), autonomic nervous system dysregulation (stage IIIa), and pulmonary edema (stage IIIb). The subjects and their parents or guardians were interviewed using structured questionnaires and received comprehensive cognitive and neurologic examinations. Motor coordination, visual-motor skill, and intellectual ability were evaluated. RESULTS: Follow-up studies were conducted in 63 previously healthy children with enterovirus 71 brainstem encephalitis (49 stage II, 7 stage IIIa, and 7 stage IIIb). The mean time to follow-up was 2.8 +/- 1.0 years (range: 1.4-4.9 years). Boys outnumbered girls by 3 to 2. The mean age at diagnosis was 2.4 +/- 1.4 years (range: 0.3-7.1 years). The most common abnormal neurologic findings on admission were altered consciousness (47.6%), followed by abnormal activities of daily living (52.4%), cerebellar dysfunction (17.5%), and cranial nerve palsy (15.9%). At follow-up, 51 of 63 children had no detectable deficits. Among the remaining 12 children, 3 died during the follow-up. The remaining 9 children (14.3%) had residual deficits. Two of these with stage IIIb disease continued to have severe motor and respiratory failure. CONCLUSIONS: Residual defects were still present in a significant proportion of children with enterovirus 71 brainstem encephalitis at >2 years after their hospitalization. Children with stage II disease were most likely to have residual cerebellar defects. Those with stage IIIb disease continued to have severe respiratory and motor impairment. Long-term follow-up of this cohort is needed to determine the ultimate prognosis. PMID- 17116699 TI - Effect of opening midlevel neonatal intensive care units on the location of low birth weight births in California. AB - OBJECTIVE: Despite evidence and recommendations encouraging the delivery of high risk newborns in hospitals with subspecialty or high-level NICUs, increasing numbers are being delivered in other facilities. Causes for this are unknown. We sought to explore the impact of diffusion of specialty or midlevel NICUs on the types of hospitals in which low birth weight newborns are born. DESIGN: We used birth certificate, death certificate, and hospital discharge data for essentially all low birth weight, singleton California newborns born between 1993 and 2000. We identified areas likely to have been affected by the opening of a new nearby midlevel unit, analyzed changes over time in the share of births that took place in midlevel NICU hospitals, and compared patterns in areas that were and were not likely affected by the opening of a new midlevel unit. We also tracked the corresponding changes in the share of births in high-level hospitals and in those without NICU facilities (low-level). RESULTS: The probability of a 500- to 1499-g infant being born in a midlevel unit increased by 17 percentage points after the opening of a new nearby unit. More than three quarters of this increase was accounted for by reductions in the probability of birth in a hospital with a high level unit (-15 points), and the other portion was resulting from reductions in the share of newborns delivered in hospitals with low-level centers (-2 points). Similar patterns were observed in 1500- to 2499-g newborns. CONCLUSIONS: The introduction of new midlevel units was associated with significant shifts of births from both high-level and low-level hospitals to midlevel hospitals. In areas in which new midlevel units opened, the majority of the increase in midlevel deliveries was attributable to shifts from high-level unit births. Continued proliferation of midlevel units should be carefully assessed. PMID- 17116700 TI - Impact of prenatal chlorpyrifos exposure on neurodevelopment in the first 3 years of life among inner-city children. AB - OBJECTIVE: The purpose of this study was to investigate the impact of prenatal exposure to chlorpyrifos on 3-year neurodevelopment and behavior in a sample of inner-city minority children. METHODS: As part of an ongoing prospective cohort study in an inner-city minority population, neurotoxicant effects of prenatal exposure to chlorpyrifos were evaluated in 254 children through the first 3 years of life. This report examined cognitive and motor development at 12, 24, and 36 months (measured with the Bayley Scales of Infant Development II) and child behavior at 36 months (measured with the Child Behavior Checklist) as a function of chlorpyrifos levels in umbilical cord plasma. RESULTS: Highly exposed children (chlorpyrifos levels of >6.17 pg/g plasma) scored, on average, 6.5 points lower on the Bayley Psychomotor Development Index and 3.3 points lower on the Bayley Mental Development Index at 3 years of age compared with those with lower levels of exposure. Children exposed to higher, compared with lower, chlorpyrifos levels were also significantly more likely to experience Psychomotor Development Index and Mental Development Index delays, attention problems, attention deficit/hyperactivity disorder problems, and pervasive developmental disorder problems at 3 years of age. CONCLUSIONS: The adjusted mean 36-month Psychomotor Development Index and Mental Development Index scores of the highly and lower exposed groups differed by only 7.1 and 3.0 points, respectively, but the proportion of delayed children in the high-exposure group, compared with the low exposure group, was 5 times greater for the Psychomotor Development Index and 2.4 times greater for the Mental Development Index, increasing the number of children possibly needing early intervention services. PMID- 17116701 TI - Pref-1, a preadipocyte secreted factor that inhibits adipogenesis. AB - Preadipocyte factor 1 (Pref-1) belongs to the Notch/Delta/Serrate family of epidermal growth factor-like repeat-containing proteins. Pref-1 is highly expressed in 3T3-L1 cells but is extinguished during adipocyte differentiation. Pref-1 serves as an excellent marker for preadipocytes. Furthermore, Pref-1 is an inhibitor of adipogenesis. Constitutive expression of Pref-1 inhibits, whereas antisense Pref-1 enhances, 3T3-L1 adipocyte differentiation. We found that Pref-1 is synthesized as a transmembrane protein but processed to generate soluble forms, including a large 50-kDa soluble form and the small soluble forms. Furthermore, only the large soluble form, but not the small soluble or the transmembrane forms of Pref-1, is biologically active to inhibit adipogenesis. We recently elucidated that the 50-kDa soluble form of Pref-1 is released by an ADAM family member, tumor necrosis factor-alpha converting enzyme (ADMA 17). In vivo, mice lacking Pref-1 show accelerated fat deposition; conversely, mice overexpressing soluble Pref-1 in adipose tissue show a decrease in fat mass, reduced expression of adipocyte markers, and lower adipocyte-secreted factors. These findings clearly demonstrate the inhibitory effect of Pref-1 on adipogenesis in vivo. PMID- 17116702 TI - History of the calorie in nutrition. AB - The calorie was not a unit of heat in the original metric system. Some histories state that a defined Calorie (modern kcal) originated with Favre and Silbermann in 1852 or Mayer in 1848. However, Nicholas Clement introduced Calories in lectures on heat engines that were given in Paris between 1819 and 1824. The Calorie was already defined in Bescherelle's 1845 Dictionnaire National. In 1863, the word entered the English language through translation of Ganot's popular French physics text, which defined a Calorie as the heat needed to raise the temperature of 1 kg of water from 0 to 1 degrees C. Berthelot distinguished between g- and kg-calories by 1879, and Raymond used the kcal in a discussion of human energy needs in an 1894 medical physiology text. The capitalized Calorie as used to indicate 1 kcal on U.S. food labels derives from Atwater's 1887 article on food energy in Century magazine and Farmers' Bulletin 23 in 1894. Formal recognition began in 1896 when the g-calorie was defined as a secondary unit of energy in the cm-g-s measurement system. The thermal calorie was not fully defined until the 20th century, by which time the nutritional Calorie was embedded in U.S. popular culture and nutritional policy. PMID- 17116703 TI - The National Research Council's assessment of research doctorate programs can be used to strengthen doctoral programs in nutrition. AB - The National Research Council (NRC) is for the first time including the field of nutrition in its Assessment of Research Doctorate Programs in 2006. This assessment will rate doctoral programs, in terms of research impact and graduate student support and outcomes, through the use of questionnaires and standardized national databases (such as research funding, publications, and citations of publications) rather than through name recognition as was used in past NRC surveys of graduate programs. Nutritionists can make this survey more valuable to the field by making sure all eligible faculty involved in training of graduate students in nutrition are included in the survey, by encouraging all eligible faculty members to complete the faculty questionnaire, and by being prepared to use and discuss the data and reports when they are released in 2007. The nutrition community should use the data from this national survey to strengthen doctoral programs and research in nutrition. PMID- 17116704 TI - Docosahexaenoic acid inhibits adipocyte differentiation and induces apoptosis in 3T3-L1 preadipocytes. AB - Docosahexaenoic acid (DHA, C22:6), a (n-3) fatty acid in fish oil, has been shown to decrease body fat and fat accumulation in rodents. We investigated the direct effect of DHA on cell growth, differentiation, apoptosis, and lipolysis using 3T3 L1 adipocytes. Cells were treated with 25-200 mumol/L DHA containing 0.2 mmol/L alpha-tocopherol or bovine serum albumin vehicle as a control. Proliferation of preconfluent preadipocytes was not affected by the DHA treatment. When added to postconfluent preadipocytes, all concentrations of DHA inhibited differentiation associated mitotic clonal expansion (P < 0.01). Postconfluent preadipocytes demonstrated apoptosis after 48 h with 100 mumol/L DHA and after 24 and 48 h with 200 mumol/L DHA (P < 0.01). Differentiation was examined by Oil Red O staining and glycerol-3-phosphate dehydrogenase (GPDH) activity after DHA treatment for 6 d. DHA decreased mean droplet size and percent lipid area in a dose-dependent manner (P < 0.01). GPDH activity was also decreased by DHA treatment (P < 0.01). In fully differentiated adipocytes, DHA increased basal lipolysis compared with the control (P < 0.01). These results demonstrate that DHA may exert its antiobesity effect by inhibiting differentiation to adipocytes, inducing apoptosis in postconfluent preadipocytes and promoting lipolysis. PMID- 17116705 TI - Chili, but not turmeric, inhibits iron absorption in young women from an iron fortified composite meal. AB - Chili and turmeric are common spices in indigenous diets in tropical regions. Being rich in phenolic compounds, they would be expected to bind iron (Fe)(3) in the intestine and inhibit Fe absorption in humans. Three experiments were conducted in healthy young women (n = 10/study) to assess the effect of chili and turmeric on Fe absorption from a rice-based meal containing vegetables and iron fortified fish sauce in vivo. Iron absorption was determined by erythrocyte incorporation of stable isotope labels ((57)Fe/(58)Fe) using a randomized crossover design. Addition of freeze-dried chili (4.2 g dry powder, 25 mg polyphenols as gallic acid equivalents) reduced Fe absorption from the meal by 38% (6.0% with chili vs. 9.7% without chili, P = 0.0017). Turmeric (0.5 g dry powder, 50 mg polyphenols as gallic acid equivalents) did not inhibit iron absorption (P = 0.91). A possible effect of chili on gastric acid secretion was indirectly assessed by comparing Fe absorption from acid soluble [(57)Fe]-ferric pyrophosphate relative to water soluble [(58)Fe]-ferrous sulfate from the same meal in the presence and absence of chili. Chili did not enhance gastric acid secretion. Relative Fe bioavailability of ferric pyrophosphate was 5.4% in presence of chili and 6.4% in absence of chili (P = 0.47). Despite the much higher amount of phenolics in the turmeric meal, it did not affect iron absorption. We conclude that both phenol quality and quantity determine the inhibitory effect of phenolic compounds on iron absorption. PMID- 17116706 TI - Differential biliary excretion of genistein metabolites following intraduodenal and intravenous infusion of genistin in female rats. AB - The purpose of this study was to determine whether bioflavonoid glucoside O conjugates are absorbed from the intestine in the intact form or as their aglycones following hydrolysis by intestinal beta-glucosidases. In this study, the intestinal absorption of genistin, the beta-glucoside of the isoflavone genistein, was examined in anesthetized, adult female rats fitted with indwelling biliary cannulas. To first establish whether genistein, once absorbed, was converted into unique metabolites, genistin was infused into the femoral or portal veins and bile samples quantitatively collected. Analysis of bile samples by HPLC-mass spectrometry revealed that almost full recovery of the genistein component occurred in the form of unreacted genistin ( approximately 20%) and genistein 7beta-O-glucuronide ( approximately 80%). However, when genistin was infused into the upper small intestine, only genistein 7beta-O-glucuronide and the aglycone genistein appeared in the bile. There was no evidence for any biliary secretion of the unreacted genistin, thereby excluding its uptake in the intact form from the small intestine in this animal model. PMID- 17116707 TI - Adipose fatty acid composition and rate of incorporation of alpha-linolenic acid differ between normal and lipoprotein lipase-deficient cats. AB - Normal adiposity occurs in humans and mice deficient of adipose lipoprotein lipase (LPL) activity. Subnormal adiposity found in LPL-deficient cats is indicative of limited de novo synthesis of fatty acids (FAs). In 14 LPL-deficient (3.0 +/- 0.1 kg) and 8 normal (3.7 +/- 0.1 kg) queens, FAs in triacylglycerol (TAG), phospholipid (PL), and nonesterified FAs (NEFAs) of plasma and inguinal subcutaneous adipose were determined before and after (d 38, 61, 110, 117, and 251) dietary linseed oil supplementation (30 g/kg). By d 60, LPL-deficient queens gained body weight (+0.4 +/- 0.1 kg), developed normal body fat mass (25 +/- 2%), and were enriched in 18:3(n-3) in their plasma and adipose lipids. Adipose TAG 18:3(n-3) enrichment in LPL-deficient queens was subnormal at all sampling times and, as observed in normal queens, apparently not equilibrated by d 251. Adipose FA profiles in TAG but not PL were substantially different (P < 0.05) between LPL deficient and normal queens; the 16:0 to 18:2(n-6) ratio was high in LPL deficient (2.4-4.4) relative to normal queens (1.0-1.4). In LPL-deficient queens, fed-state plasma NEFA (n-6) and (n-3) enrichments were similar to those in adipose TAG, and plasma NEFA concentration was high (0.62 +/- 0.05 mmol/L) and similar to that in normal queens after withholding diet for 16 h. These data indicate that LPL deficiency in cats reduces dietary FA storage efficiency, favors storage of saturated over unsaturated FAs, and stimulates de novo FA synthesis substantive enough to support normal adiposity. PMID- 17116708 TI - Almonds decrease postprandial glycemia, insulinemia, and oxidative damage in healthy individuals. AB - Strategies that decrease postprandial glucose excursions, including digestive enzyme inhibition, and low glycemic index diets result in lower diabetes incidence and coronary heart disease (CHD) risk, possibly through lower postprandial oxidative damage to lipids and proteins. We therefore assessed the effect of decreasing postprandial glucose excursions on measures of oxidative damage. Fifteen healthy subjects ate 2 bread control meals and 3 test meals: almonds and bread; parboiled rice; and instant mashed potatoes, balanced in carbohydrate, fat, and protein, using butter and cheese. We obtained blood samples at baseline and for 4 h postprandially. Glycemic indices for the rice (38 +/- 6) and almond meals (55 +/- 7) were less than for the potato meal (94 +/- 11) (P < 0.003), as were the postprandial areas under the insulin concentration time curve (P < 0.001). No postmeal treatment differences were seen in total antioxidant capacity. However, the serum protein thiol concentration increased following the almond meal (15 +/- 14 mmol/L), indicating less oxidative protein damage, and decreased after the control bread, rice, and potato meals (-10 +/- 8 mmol/L), when data from these 3 meals were pooled (P = 0.021). The change in protein thiols was also negatively related to the postprandial incremental peak glucose (r = -0.29, n = 60 observations, P = 0.026) and peak insulin responses (r = -0.26, n = 60 observations, P = 0.046). Therefore, lowering postprandial glucose excursions may decrease the risk of oxidative damage to proteins. Almonds are likely to lower this risk by decreasing the glycemic excursion and by providing antioxidants. These actions may relate to mechanisms by which nuts are associated with a decreased risk of CHD. PMID- 17116709 TI - Accelerated transferrin degradation in HFE-deficient mice is associated with increased transferrin saturation. AB - HFE, a major histocompatibility complex class I-related protein, is implicated in the iron overload disease, hereditary hemochromatosis. Whereas patients with hereditary hemochromatosis have low serum transferrin levels, little is known about transferrin turnover in HFE deficiency states. We injected mice intravenously with radioiodinated transferrin and compared plasma transferrin decay and steady-state endogenous transferrin concentration in the plasma between HFE-deficient and wild-type C57BL/6 mouse strains. HFE-deficient mice degraded transferrin faster than normal (P < 0.001) and had lower plasma transferrin concentrations (P < 0.001). Both HFE-deficient and wild-type mice were then fed diets with 3 different iron concentrations that we designated deficient (2-5 mg/kg of iron), control (0.2 g/kg), and overload (20 g/kg) for 6 wk immediately after weaning to create a range of serum iron concentrations and resultant transferrin saturations ranging from 16 to 78%. We found an inverse correlation between transferrin saturation and transferrin half-life (P < 0.0001, r = -0.839) for both HFE-deficient and wild-type mice, which suggests that HFE does not have a direct effect on transferrin catabolism; rather, HFE may influence transferrin half-life indirectly through its effect on transferrin saturation, which in turn enhances transferrin decay in HFE-deficient mice. PMID- 17116710 TI - Pinto beans are a source of highly bioavailable copper in rats. AB - The trace element copper (Cu) is a required nutrient in the diets of humans. It has been found in animal studies to be essential for efficient iron absorption and oxygen utilization and for aiding free-radical degradation. Dry beans (Phaseolis vulgaris) are potentially good sources of Cu; thus, the objective of this study was to determine the bioavailability of Cu from dry beans using the pinto bean as the source. Dry beans were obtained from a local market, cooked according to package directions, and dried. Weanling male Sprague-Dawley rats (8 groups of 8 rats each) were fed a Cu-deficient diet (AIN-93G) for 4 wk followed by 2 wk of Cu repletion with diets containing 0-6.5 mg Cu/kg diet added as CuSO(4) or with 0.6 and 1.5 mg Cu/kg incorporated into rat diets as pinto beans at 10 and 20%. Standard response curves were developed based on repletion-induced recovery of 10 indices of Cu status, including organ Cu concentrations and Cu dependent enzyme activities, in response to increasing dietary Cu as CuSO(4). Recovery of these variables in rats fed the pinto bean diets was compared with the standard response curve at similar levels of dietary Cu. Based on the recovery of all 10 variables, the relative bioavailability of Cu from dry beans was at least 100% of that with the highly available CuSO(4). For 3 of the variables, liver and heart Cu concentrations and serum superoxide dismutase 3 activity, estimated bioavailability values of Cu from beans were 138, 140, and 134%, respectively, of those from CuSO(4). We conclude that the dry pinto bean is a good source of dietary Cu with respect to both concentration and bioavailability. PMID- 17116711 TI - Hepatic phosphatidylethanolamine N-methyltransferase expression is increased in diabetic rats. AB - Phosphatidylcholine is an essential phospholipid that is synthesized by 2 different pathways, the CDP-choline pathway and the methylation of phosphatidylethanolamine by phosphatidylethanolamine N-methyltransferase (PEMT). Recent studies have suggested that PEMT is an important consumer of methyl groups from S-adenosylmethionine (SAM) and is a major determinant of homocysteine pools. Diabetes and all-trans-retinoic acid (ATRA) have been shown to alter the activities of several enzymes involved in methyl group metabolism. Thus, we investigated how diabetes and ATRA, individually and together, affect SAM dependent phospholipid methylation. Rats received a single injection of streptozotocin (60 mg/kg body wt) or vehicle followed by administration of ATRA (30 mumol/kg body wt) or vehicle for 5 d. The hepatic activity of PEMT increased 50% in both diabetic rat groups, whereas administration of ATRA was without effect. In diabetic rats, plasma total homocysteine decreased 30-35% in all treatment groups as compared with the control group. Thus, alterations in the activity of PEMT were not directly correlated to changes in homocysteine concentrations. Moreover, treatment of diabetic rats with insulin prevented the increase in PEMT activity and abundance. Because these observations support an increased need for SAM-dependent phosphatidylcholine synthesis, this may also indicate an increased choline requirement in diabetes. PMID- 17116712 TI - Belgrade rats display liver iron loading. AB - Patients with mutations in divalent metal transporter-1 (DMT1), an intestinal nonheme iron transporter, suffer from microcytic anemia and hepatic iron loading. DMT1 is also mutated in Belgrade rats, an animal model with a thalassemic-like disorder of microcytic anemia with hyperferrinemia. However, aspects of hepatic iron loading in this genetic model are not well characterized. To more fully define the Belgrade rat's iron status, we compared the characteristics of homozygous (b/b) and heterozygous (b/+) rats fed an iron-supplemented diet for 3 wk postweaning. Dietary supplementation with ferrous iron improved the anemia of b/b rats insofar as hematocrits increased from 0.13 (21-d-old) to 0.31 (42-d old). However, hematocrits remained significantly lower than those of age-matched b/+ rats (0.36 and 0.41 in 21- and 42-d-old heterozygotes, respectively, P < 0.05). Wright's staining of b/b red cells confirmed the hypochromic microcytic nature of Belgrade rats' anemia. The liver iron concentration of 42-d-old b/b rats was greater than in age-matched b/+ rats (5.97 vs. 2.24 mumol/g, P < 0.05). Whereas Perls' Prussian blue iron staining was evident in both periportal and centrilobular regions in 42-d-old b/b liver sections, no staining was observed in age-matched b/+ tissue sections. Quantitative real-time PCR analysis showed that expression of liver hepcidin mRNA in 42-d-old b/b rats was 3-fold greater than age-matched b/+ rats. These results indicate that, similar to human patients with DMT1 mutations, Belgrade rats also display hepatic iron loading. Our data suggest this condition arises from ineffective erythropoiesis. PMID- 17116713 TI - Dietary folate and APC mutations in sporadic colorectal cancer. AB - Folate deficiency has been associated with colorectal cancer risk and may be involved in colorectal carcinogenesis through increased chromosome instability, gene mutations, and aberrant DNA methylation. Within the Netherlands Cohort Study on diet and cancer, we investigated the associations between dietary folate intake and colorectal cancer risk with (APC(+)) and without (APC(-)) truncating APC mutations, accounting for hMLH1 expression and K-ras mutations. In total, 528 cases and 4200 subcohort members were available for data analyses of the study cohort (n = 120,852) from a follow-up period between 2.3 and 7.3 y after baseline. Adjusted gender-specific incidence rate ratios (RR) over tertiles of folate intake were calculated in case-cohort analyses for colon and rectal cancer. Although relatively high folate intake was not associated with overall colorectal cancer risk, it reduced the risk of APC(-)colon tumors in men (RR 0.58, 95% CI 0.32-1.05, P(trend) = 0.06 for the highest vs. lowest tertile of folate intake). In contrast, it was positively associated with APC(+) colon tumors in men (highest vs. lowest tertile: RR 2.77, 95% CI 1.29-5.95, P(trend) = 0.008) and was even stronger when the lack of hMLH1 expression and K-ras mutations were excluded (RR 3.99, 95% CI 1.43-11.14, P(trend) = 0.007). Such positive associations were not observed among women; nor was folate intake associated with rectal cancer when APC mutation status was taken into account. Relatively high folate consumption reduced the risk of APC(-) colon tumors, but folate intake was positively associated with APC(+) colon tumors among men. These opposite results may indicate that folate enhances colorectal carcinogenesis through a distinct APC mutated pathway. PMID- 17116714 TI - Systemic oxidative alterations are associated with visceral adiposity and liver steatosis in patients with metabolic syndrome. AB - Although evidence suggests the link between chronic inflammation and oxidative stress as the main mechanism responsible for endothelial dysfunction and cardiovascular complications in patients with metabolic syndrome, little is known about the determining role of each metabolic syndrome component in such alterations. This study investigated the relation between systemic oxidative alterations and metabolic syndrome features in 41 patients. Compared with control subjects, serum vitamin C and alpha-tocopherol concentrations were lower and those of lipid peroxides [thiobarbituric acid reactive substances (TBARs)] were higher in metabolic syndrome patients (P < 0.001). A linear relation was observed between visceral fat thickness and serum TBARs:cholesterol ratio (r = 0.541, P < 0.001), whereas negative correlations were found between alpha-tocopherol and BMI (r = -0.212, P < 0.05) and the grade of liver steatosis (r = -0.263, P < 0.02). Patients with metabolic syndrome and liver steatosis had higher serum hyaluronate (HA) concentrations (P < 0.001). Serum HA was positively correlated with serum alanine amino transferase (r = 0.715, P < 0.001) and the homeostasis monitoring assessment index (r = 0.248, P < 0.03). The presence of metabolic syndrome was predicted from a linear combination of visceral fat and all oxidative variables. In metabolic syndrome patients, serum nitrosothiols and vitamin C concentrations, which were lower (P < 0.001) than in control subjects, were inversely related to the grade of hypertension (r = -0.645, P < 0.001 and r = -0.415, P < 0.007, respectively). In conclusion, metabolic syndrome patients exhibited decreased antioxidant protection and increased lipid peroxidation. Our results indicate a strong association between increased abdominal fat storage, liver steatosis, and systemic oxidative alterations in metabolic syndrome patients and diminished nitrosothiols and vitamin C concentrations as important factors associated with hypertension in these patients. PMID- 17116715 TI - Moderate alcohol consumption is more cardioprotective in men with the metabolic syndrome. AB - The aim of the present study was to evaluate the relation among alcohol consumption, the metabolic syndrome, and the risk of ischemic heart disease (IHD). The study was conducted in a cohort of 1966 men from the Quebec Cardiovascular Study. All men were initially free of IHD and, during the follow up period of 13 y, 219 first cases of IHD were diagnosed. Alcohol consumption was determined by calculating the g/d intake based on standard portions of beer, wine, and spirits. Metabolic syndrome was diagnosed according to a modification of the National Cholesterol Education Program Adult Treatment Panel III definition. Men who consumed >or=15.2 g of alcohol/d (4th quartile of the distribution) were younger (P < 0.001), had elevated plasma HDL-C concentrations (P < 0.001), and lower plasma concentrations of insulin (P = 0.01), CRP (P = 0.01), and fibrinogen (P < 0.001) than men in the 1st quartile (<1.3 g of alcohol/d). After adjustment for a series of coronary risk factors, alcohol consumption >or=15.2 g/d was associated with a 39% reduction in the 13-y risk of IHD [relative risk (RR) of IHD = 0.61, P = 0.02]. Finally, an alcohol consumption <15.2 g/d was associated with an increase of the risk of IHD in men with the metabolic syndrome (RR = 2.24, P < 0.001) but not in men without the metabolic syndrome (RR = 1.31, P = 0.22). These results confirm that moderate daily alcohol consumption has cardioprotective properties and suggest that the effects may be more important in subjects with a deteriorated risk profile, such as those with the metabolic syndrome. PMID- 17116716 TI - Supplemental dietary inulin affects the bioavailability of iron in corn and soybean meal to young pigs. AB - Iron deficiency represents one of the most common global nutritional disorders in humans. Our objective was to determine whether and how supplemental inulin improved utilization of iron intrinsically present in a corn and soybean meal diet by young pigs for hemoglobin repletion. In Expt. 1, 3 groups (n = 8/group) of pigs were fed a corn and soybean meal-based diet (BD, without inorganic iron addition) or BD + 2 or 4% inulin (Synergy 1: a mixture of oligofructose and long chain inulin HP, Orafti) for 5 wk. Final blood hemoglobin concentrations and the overall hemoglobin repletion efficiency of pigs were positively (r = 0.55 and 0.69, P < 0.01) correlated with dietary inulin concentrations. Compared with pigs fed the BD, those fed 4% inulin demonstrated a 28% improvement (P < 0.01) in hemoglobin repletion efficiency and 15% (P < 0.01) improvement in the final blood hemoglobin concentration. In Expt. 2, 12 weanling pigs (n = 6/group) were fed the BD or the BD + 4% inulin for 6 wk. Pigs fed 4% inulin had higher (P < 0.05) soluble Fe concentrations in the digesta of the proximal, mid, and distal colon, and lower (P < 0.05) sulfide concentrations in the digesta of the distal colon. Supplemental inulin had virtually no effect on pH or phytase activity of digesta from any of the tested segments. In conclusion, supplementing 4% inulin improved utilization of intrinsic iron in the corn and soybean meal diet by young pigs, and this benefit was associated with soluble Fe and sulfide concentrations but not pH or phytase activity in the digesta. PMID- 17116717 TI - Dietary flavonoids and flavonoid-rich foods are not associated with risk of type 2 diabetes in postmenopausal women. AB - Flavonoids have anti-inflammatory and antioxidative effects and thus may protect against diabetes. Therefore, we hypothesized that consumption of flavonoids and specific food and beverage sources of flavonoids would be associated with reduced risk of incident diabetes. At baseline (1986), diet (by food frequency questionnaire) and health information were collected from 35,816 postmenopausal women free of diabetes. Self-reported incident diabetes was ascertained 5 times during the study (1987, 1989, 1992, 1997, and 2004). Cox proportional hazards regression was used to calculate hazard ratios for incident diabetes according to categories of total flavonoids and anthocyanidins, flavones, flavanones, flavonols, flavan-3-ol monomers, isoflavones, and proanthocyanidins. Hazard ratios according to intake categories of flavonoid-rich foods and beverages were also calculated (apples, pears, berries, broccoli, bran, citrus, tea, and red wine). Flavonoid consumption was not associated with diabetes risk after multivariable adjustment. Although other flavonoid-rich foods and beverages were not associated, red wine was inversely associated with diabetes. Women who reported drinking red wine >or=1 time/wk had a 16% reduced risk of diabetes than those drinking wine <1 time/wk [HR (95% CI): 0.84 (0.71, 0.99)], with parallel findings for white wine, beer, and liquor. In conclusion, these data do not support a diabetes-protective effect of flavonoids. The suggestive evidence of a protective effect of regular red wine consumption is shared with an inverse association between alcohol drinks in general and diabetes risk and may reflect the effects of nonflavonoid constituents that are common to all alcohol drinks. PMID- 17116719 TI - Carotenoid and tocopherol estimates from the NCI diet history questionnaire are valid compared with multiple recalls and serum biomarkers. AB - To improve the measurement of usual dietary intake, the National Cancer Institute developed a cognitively based Diet History Questionnaire (DHQ), which has been validated against four 24-h dietary recalls (4 24-HR) for energy, macronutrients, and several vitamins and minerals. This analysis used data from The Eating at America's Table Study (EATS) to determine the validity of estimates for carotenoids and tocopherols from the DHQ. Over the course of a year, 163 participants provided 1 or 2 blood samples and completed the DHQ and 4 24-HR. For both the DHQ and the 4 24-HR, crude correlations between serum and diet were modest to strong for the provitamin A carotenoids (alpha-carotene, beta-carotene, beta-cryptoxanthin), low to modest for lycopene, and very low for lutein. The individual dietary tocopherols were weakly correlated with the serum tocopherols, but vitamin E from food and dietary supplements was strongly and positively correlated with serum alpha-tocopherol and strongly and inversely correlated with serum gamma-tocopherol for both instruments. Adjustment for energy, BMI, smoking status, serum total cholesterol, and serum triacylglycerol did not appreciably change the correlations. Using the method of triads, validity coefficients for the DHQ were comparable to the 4 24-HR and were especially strong for alpha carotene, beta-cryptoxanthin, lutein + zeaxanthin, and total vitamin E in men and gamma-tocopherol and total vitamin E in women. In this study, there was no advantage of 2 blood samples over 1, suggesting reasonably stable ranking of individuals for these biomarkers, which is important for large epidemiologic studies that typically obtain only 1 blood sample for biomarker status. PMID- 17116718 TI - Dietary phytoestrogen intake is associated with reduced colorectal cancer risk. AB - Evidence suggests dietary phytoestrogens may reduce the risk of certain hormonal cancers (e.g. breast and prostate). There is a paucity of data regarding phytoestrogens and colorectal cancer risk. Phytoestrogens are plant compounds with estrogen-like activities. Main classes include isoflavones (found in legumes such as soy) and lignans (found in grains, seeds, nuts, fruits, and vegetables). Although isoflavones have dominated phytoestrogen cancer research, lignans may be more relevant to North American diets. Food questionnaires and analytic databases have recently been modified to incorporate some lignan information. We conducted a case-control study to evaluate the association between phytoestrogen intake and colorectal cancer risk. Colorectal cancer cases were diagnosed in 1997-2000, aged 20-74 y, identified through the population-based Ontario Cancer Registry, and recruited by the Ontario Familial Colorectal Cancer Registry. Controls were a sex and age-group matched random sample of the population of Ontario. Epidemiologic and food frequency questionnaires were completed by 1095 cases and 1890 control subjects. Multivariate logistic regression analysis was used to obtain adjusted odds ratio (OR) estimates. Dietary lignan intake was associated with a significant reduction in colorectal cancer risk [OR (T3 vs. T1) = 0.73; 95% CI: 0.56, 0.94], as was isoflavone intake [OR (T3 vs. T1) = 0.71; 95% CI: 0.58, 0.86]. We evaluated interactions between polymorphic genes that encode enzymes possibly involved in metabolism of phytoestrogens (CYPs, catechol O-methyl transferase, GSTs, and UGTs) and found no significant effect modification with respect to phytoestrogen intake. This finding that phytoestrogen intake may reduce colorectal cancer risk is important, because dietary intake is potentially modifiable. PMID- 17116720 TI - Gender differences in body mass index in rural India are determined by socio economic factors and lifestyle. AB - A survey of the nutritional status of women in 6 villages of the Pune district of Maharashtra, India, found young women to have a significantly lower BMI than their male peers. The purpose of this study was to identify social and economic factors associated with this difference in thinness and to explore the behavior in men and women that might underlie these associations. We compared men and women in 90 families in this part of Maharashtra by taking measurements of the height and weight of the married couple of child-bearing age in each family and assessing their social and economic details, fasting practices, and oil consumption. In this agricultural community, women were thinner in joint land owning families, where the main occupation was farming, than those in nonfarming families. This was not true of men in this type of family. Men in "cash-rich" families had higher BMI than men in families without this characteristic. There was no corresponding difference in women's BMI. We then examined the lifestyles of men and women in a subset of 45 of these families. Women were more likely to work full time in farming than men, to carry the burden of all household chores, to have less sleep, and to eat less food away from home than men. Women fasted more frequently and more strictly than men. Despite identifying significant differences in behavior between men and women in the same household, we did not find a direct link between behavior and BMI. We conclude that being married into a farming family is an important factor in determining the thinness of a woman in rural Maharashtra. PMID- 17116721 TI - Daily intake of heat-killed Lactobacillus plantarum L-137 augments acquired immunity in healthy adults. AB - Heat-killed Lactobacillus plantarum strain L-137 (HK-LP) is a potent inducer of IL-12 in vitro as well as in vivo in mice. HK-LP has been shown to suppress IgE production against food allergens, as well as tumor growth in mice, through IL-12 production, which induces the T helper (Th) 1 type immune response. To determine whether the intake of HK-LP influences immune function and the quality of life (QOL), a randomized, double-blind, placebo-controlled, parallel study was conducted in healthy subjects. Sixty subjects (30 men and 30 women, mean age 56.3 y) were randomly assigned to receive a capsule containing 10 mg of HK-LP daily or a matching capsule for 12 wk. Biomarkers for innate immunity such as the natural killer activity of peripheral blood mononuclear cells, neutrophil phagocytosis, and cell surface expression of CD64 on monocytes were measured every 4 wk. Biomarkers for acquired immunity such as concanavalin A (Con A)-induced proliferation, percentages of INF-gamma and IL-4-producing cluster of differentiation (CD)4(+) T cells (Th1:Th2 ratio), and the serum IgG4:IgG ratio were measured every 4 wk or at wk 0 and wk 12. Health-related QOL was assessed using a self-rating questionnaire with 26 items. Among the measured biomarkers, the percent change in Con A-induced proliferation and the Th1:Th2 ratio in the HK LP group was greater than those in the control group (P = 0.036 and P = 0.002, respectively). The degree of improvement in QOL was higher in the HK-LP group than in the control group at wk 8 (P = 0.049) and tended to be higher at wk 12 (P = 0.092). These results suggest that a daily intake of HK-LP augments acquired immunity, especially Th1-related immune functions in healthy subjects, thereby improving the health-related QOL. PMID- 17116722 TI - Predicted serum folate concentrations based on in vitro studies and kinetic modeling are consistent with measured folate concentrations in humans. AB - The nutritional quality of new functional or fortified food products depends on the bioavailability of the nutrient(s) in the human body. Bioavailability is often determined in human intervention studies by measurements of plasma or serum profiles over a certain time period. These studies are time and cost consuming and often appear to lack an optimal study design, leading to follow-up intervention trials. Therefore, an alternative approach is needed that will optimize the development of new products. This study describes an approach to predict human serum concentrations after the consumption of (fortified) food products. The concept is based on the integration of in vitro results with kinetic modeling. As a case study, human serum folate concentrations were predicted after the consumption of folate-fortified milk products for 4 wk. Oral bioavailability was investigated using a step-wise approach in which luminal bioaccessibility and intestinal absorption were independently evaluated. Subsequently, these in vitro data were integrated in a kinetic mathematical (in silico) model to predict serum folate concentrations after the intake of a single dose and during long-term consumption. This approach was evaluated in comparison to a human intervention study in which folic acid-fortified milk products were tested for their effect on serum folate concentrations. A high predictive quality of this alternative in vitro/in silico approach was demonstrated. Finally, this methodology was applied to predict serum folate concentrations after intake of different fortified milk products for 4 wk, showing its benefits for the development of new nutritional products. PMID- 17116723 TI - Use of the affinity/HPLC method for quantitative estimation of folic acid in enriched cereal-grain products. AB - In 1998, the United States introduced mandatory fortification of enriched cereal grain products with folic acid to reduce the incidence of neural tube defects. As a consequence, substantial amounts of folic acid, the synthetic form of folate, were added to the American diet, and the ability to assess folic acid intake took on greater importance. The purpose of the current study was to separate and quantify folic acid and 5-methyltetrahydrofolate, the most prominent naturally occurring folate in fortified foods, with a reliable and robust method. Folates were heat-extracted from food samples. A trienzyme treatment (alpha-amylase, rat plasma conjugase, and protease) was applied to the extracts followed by purification by affinity chromatography. Folic acid and 5-methyltetrahydrofolate were separated and quantified by reversed-phase HPLC with fluorescence and UV detection. A gradient elution with phosphate buffer and acetonitrile was used to separate the different forms of folates. The method gave a linear response in a range of 0.1-3 mumol/L and 0.0125-0.25 mumol/L for folic acid and 5 methyltetrahydrofolate, respectively. These ranges were similar to the expected levels in the samples. The CV of the peak areas of folic acid and 5 methyltetrahydrofolate for 5 commercial wheat flour samples extracted and run separately on the same day was 2.0 and 5.7% and, run over 5 consecutive days, was 7.2 and 7.3%, respectively. Total folate values in 45 samples of fortified food measured by HPLC and by the traditional microbiological assay demonstrated a high correlation (r(2) = 0.986). PMID- 17116724 TI - Role of O6-methylguanine-DNA methyltransferase in protecting from alkylating agent-induced toxicity and mutations in mice. AB - The DNA repair protein O(6)-methylguanine-DNA methyltransferase (MGMT) protects from toxicity and mutations incurred following alkylating agents by removing O(6) alkylguanine lesions. Using Mgmt-/- mice, we examined MGMT's role in protecting from in vivo mutations induced by three different alkylating agents, temozolomide (TMZ), 1,3-bis (2-chloroethyl)-1-nitrosourea (BCNU) and cyclophosphamide. Mutant frequencies were determined in the hypoxanthine-guanine phosphoribosyltransferase gene of splenic T-lymphocytes from C57BL/6 mice (Mgmt+/+ and Mgmt-/-) following TMZ, BCNU or cyclophosphamide. Following TMZ, the mutation frequency was significantly greater in Mgmt-/- mice (5.5 and 9.8 x 10(-6) for 7 and 10 mg/kg TMZ, respectively) compared with vehicle-treated mice (1.0 x 10(-6), P 6 months of age were compared with healthy wild-type (WT) controls. Inflammation was assessed using macroscopic/histological scores and myeloperoxidase activity as an indication of granulocyte infiltration. Mucosal polyps were scored macroscopically and hyperproliferation was quantified by 5-bromo-2-deoxyuridine immunohistochemistry. Dysplastic changes were assessed histologically based on cytologic and architectured atypia as well as the presence of submucosal invasion. The p53 and beta-catenin messenger RNA mRNA and protein expression were assessed using real time polymerase chain reaction and immunohistochemistry, respectively. Inflammatory indices were significantly elevated in interleukin-10-deficient (IL 10(-/-)) over WT and not significantly different from IL-10(-/-)/iNOS(-/-) mice. The incidence of mucosal polyps was similar between IL-10(-/-) (79%) and IL-10(-/ )/iNOS(-/-) (83%) mice; however, significantly higher numbers of polyps were observed in the absence of iNOS (P < 0.05). Hyperproliferation was noted in both groups. Signs of dysplasia and submucosal invasion were significantly higher in IL-10(-/-)/iNOS(-/-) compared with IL-10(-/-) mice (P < 0.05). No significant increase in p53 and beta-catenin mRNA levels was observed in IL-10(-/-) over WT mice; however, a 2-fold (P = 0.06) and 3-fold (P < 0.05) increase, respectively, was noted in IL-10(-/-)/iNOS(-/-) mice. Our data suggest exposure to chronic NO limits abnormal p53 and beta-catenin expression and reduces incidence of adenocarcinoma in IL-10(-/-) mice. PMID- 17116729 TI - Pulmonary arteriovenous malformation treated with embolotherapy: systemic collateral supply at multidetector CT angiography after 2-20-year follow-up. AB - PURPOSE: To retrospectively evaluate frequency of systemic arterial collateral supply to treated pulmonary arteriovenous malformations (PAVMs) in long-term follow-up with multi-detector row helical computed tomography (CT). MATERIALS AND METHODS: Institutional review board approval was obtained, with waiver of informed consent. Thirty-two patients (19 male, 13 female; mean age, 43 years) underwent follow-up multi-detector row helical CT angiography of the chest (collimation, 16x0.75 mm) 2 or more years after embolotherapy of PAVMs. The study group had a history of successful embolotherapy of 53 PAVMs and a mean of 9 years of follow-up (range, 2-20 years). A search for abnormal systemic arteries was based on analysis of thin-collimated contiguous transverse CT scans and two- and three-dimensional images including maximum intensity projections and volume rendered images. Statistical comparison was performed with the Fisher exact test (categoric variables) and Wilcoxon rank sum test (continuous variables). RESULTS: At CT, 13 patients (group 1) had abnormally enlarged systemic arteries and 19 patients (group 2) had no abnormal arteries. In group 1, 32 abnormally enlarged arteries were seen-five bronchial and 27 nonbronchial arteries (14 inferior phrenic, six musculophrenic, five internal mammary, two intercostal). The degree of enlargement was moderate for 26 arteries and marked for six. There were no significant differences between groups for (a) clinical characteristics of patients, including history of surgery before or after embolotherapy (P=.7); (b) anatomic structures of treated PAVMs; and (c) embolization procedures and their effectiveness. The number of patients with features suggestive of lung infarction in the days or months after embolotherapy was significantly higher in group 1 (P=.04). On CT angiograms, the number of patients with features suggestive of sequelae of lung infarction was significantly higher in group 1 (P=.02). There were no symptomatic differences attributable to systemic collateral supply between groups; in particular, there was no hemoptysis in group 1. CONCLUSION: Abnormally enlarged systemic arteries were present in 13 of 32 patients, in whom there was a significantly higher frequency of clinical and/or radiographic features suggestive of lung infarction after embolotherapy. PMID- 17116730 TI - Assessment of bone marrow angiogenesis in patients with acute myeloid leukemia by using contrast-enhanced MR imaging with clinically approved iron oxides: initial experience. AB - PURPOSE: To prospectively assess bone marrow (BM) angiogenesis in patients with acute myeloid leukemia (AML) by using iron oxide-enhanced magnetic resonance (MR) imaging. MATERIALS AND METHODS: The study was institutional ethics committee approved. Informed signed consent was obtained from each study participant. The requirement for informed consent for use of data from a reference database was waived. Eleven patients (seven women, four men; mean age, 53 years+/-4.40 [standard deviation]) with an initial diagnosis of AML were enrolled in the study and underwent T2*-weighted two-echo echo-planar MR imaging of the pelvis before and after intravenous injection of a clinically approved iron oxide blood-pool contrast agent. Six healthy control subjects (one woman, five men; mean age, 35 years+/-2.31) were examined with the same MR protocol. The iron oxide-induced change in R2* relaxation rate (DeltaR2*) was calculated, and the vascular volume fraction (VVF) of the BM was derived by dividing the DeltaR2* of the BM by the DeltaR2* of the muscle. Parametric DeltaR2* maps were calculated to visualize vessel distribution. Patients underwent BM biopsy for correlative determination of microvessel density (MVD) and vascular endothelial growth factor (VEGF). Differences in DeltaR2*, VVF, VEGF, and MVD were compared by using the Wilcoxon rank sum test. RESULTS: DeltaR2* maps showed prominent areas of highly vascularized BM in the patients with AML, whereas the control subjects had moderately vascularized BM with homogeneous vessel distribution. Quantitative analysis revealed VVF values to be significantly higher in patients with AML than in control subjects: The mean VVF in the pelvis was 9.18%+/-1.54 for patients versus 3.91%+/-0.61 for control subjects (P=.010). In accordance with MR results, MVD (P=.009) and VEGF expression (P=.017) were significantly elevated in the AML group compared with values in the control group. CONCLUSION: Iron oxide-enhanced MR imaging enables assessment of BM angiogenesis in patients with AML. PMID- 17116731 TI - Junctional adhesion molecule-C regulates vascular endothelial permeability by modulating VE-cadherin-mediated cell-cell contacts. AB - We recently reported that junctional adhesion molecule (JAM)-C plays a role in leukocyte transendothelial migration. Here, the role of JAM-C in vascular permeability was investigated in vitro and in vivo. As opposed to macrovascular endothelial cells that constitutively expressed JAM-C in cell-cell contacts, in quiescent microvascular endothelial cells, JAM-C localized mainly intracellularly, and was recruited to junctions upon short-term stimulation with vascular endothelial growth factor (VEGF) or histamine. Strikingly, disruption of JAM-C function decreased basal permeability and prevented the VEGF- and histamine induced increases in human dermal microvascular endothelial cell permeability in vitro and skin permeability in mice. Permeability increases are essential in angiogenesis, and JAM-C blockade reduced hyperpermeability and neovascularization in hypoxia-induced retinal angiogenesis in mice. The underlying mechanisms of the JAM-C-mediated increase in endothelial permeability were studied. JAM-C was essential for the regulation of endothelial actomyosin, as revealed by decreased F-actin, reduced myosin light chain phosphorylation, and actin stress fiber formation due to JAM-C knockdown. Moreover, the loss of JAM-C expression resulted in stabilization of VE-cadherin-mediated interendothelial adhesion in a manner dependent on the small GTPase Rap1. Together, through modulation of endothelial contractility and VE-cadherin-mediated adhesion, JAM-C helps to regulate vascular permeability and pathologic angiogenesis. PMID- 17116732 TI - An inflammation-induced mechanism for leukocyte transmigration across lymphatic vessel endothelium. AB - The exit of antigen-presenting cells and lymphocytes from inflamed skin to afferent lymph is vital for the initiation and maintenance of dermal immune responses. How such an exit is achieved and how cells transmigrate the distinct endothelium of lymphatic vessels are unknown. We show that inflammatory cytokines trigger activation of dermal lymphatic endothelial cells (LECs), leading to expression of the key leukocyte adhesion receptors intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and E-selectin, as well as a discrete panel of chemokines and other potential regulators of leukocyte transmigration. Furthermore, we show that both ICAM-1 and VCAM-1 are induced in the dermal lymphatic vessels of mice exposed to skin contact hypersensitivity where they mediate lymph node trafficking of dendritic cells (DCs) via afferent lymphatics. Lastly, we show that tumor necrosis factor alpha stimulates both DC adhesion and transmigration of dermal LEC monolayers in vitro and that the process is efficiently inhibited by ICAM-1 and VCAM-1 adhesion blocking monoclonal antibodies. These results reveal a CAM-mediated mechanism for recruiting leukocytes to the lymph nodes in inflammation and highlight the process of lymphatic transmigration as a potential new target for antiinflammatory therapy. PMID- 17116733 TI - Induction of a virus-specific effector-memory CD4+ T cell response by attenuated SIV infection. AB - We investigated simian immunodeficiency virus (SIV)-specific CD4+ T cell responses in rhesus macaques chronically infected with attenuated or pathogenic SIV strains. Analysis of SIVDeltanef-infected animals revealed a relatively high frequency of SIV-specific CD4+ T cells representing 4-10% of all CD4+ T lymphocytes directed against multiple SIV proteins. Gag-specific CD4+ T cells in wild-type SIV-infected animals were 5-10-fold lower in frequency and inversely correlated with the level of plasma viremia. SIV-specific CD4+ cells from SIVDeltanef animals were predominantly CD27-CD28-CD45RAlowCCR7-CCR5-, consistent with an effector-memory subset, and included a fully differentiated CD45RA+CCR7- subpopulation. In contrast, SIV-specific CD4+ T cells from SIV-infected animals were mostly CD27+CD28+CD45RA-CCR7+CCR5+, consistent with an early central memory phenotype. The CD45RA+CCR7-CD4+ subset from SIVDeltanef animals was highly enriched for effector CD4+ T cells, as indicated by the perforin expression and up-regulation of the lysosomal membrane protein CD107a after SIV Gag stimulation. SIV-specific CD4+ T cells in attenuated SIV-infected animals were increased in frequency in bronchioalveolar lavage and decreased in lymph nodes, consistent with an effector-memory T cell population. The ability of SIVDeltanef to induce a high frequency virus-specific CD4+ T cell response with direct effector function may play a key role in protective immunity produced by vaccination with attenuated SIV strains. PMID- 17116734 TI - Identification of a radio-resistant and cycling dermal dendritic cell population in mice and men. AB - In this study, we explored dermal dendritic cell (DC) homeostasis in mice and humans both in the steady state and after hematopoietic cell transplantation. We discovered that dermal DCs proliferate in situ in mice and human quiescent dermis. In parabiotic mice with separate organs but shared blood circulation, the majority of dermal DCs failed to be replaced by circulating precursors for >6 mo. In lethally irradiated mice injected with donor congenic bone marrow (BM) cells, a subset of recipient DCs remained in the dermis and proliferated locally throughout life. Consistent with these findings, a large proportion of recipient dermal DCs remained in patients' skin after allogeneic hematopoietic cell transplantation, despite complete donor BM chimerism. Collectively, our results oppose the traditional view that DCs are nondividing terminally differentiated cells maintained by circulating precursors and support the new paradigm that tissue DCs have local proliferative properties that control their homeostasis in the steady state. Given the role of residual host tissue DCs in transplant immune reactions, these results suggest that dermal DC homeostasis may contribute to the development of cutaneous graft-versus-host disease in clinical transplantation. PMID- 17116735 TI - Random migration precedes stable target cell interactions of tumor-infiltrating T cells. AB - The tumor microenvironment is composed of an intricate mixture of tumor and host derived cells that engage in a continuous interplay. T cells are particularly important in this context as they may recognize tumor-associated antigens and induce tumor regression. However, the precise identity of cells targeted by tumor infiltrating T lymphocytes (TILs) as well as the kinetics and anatomy of TIL target cell interactions within tumors are incompletely understood. Furthermore, the spatiotemporal conditions of TIL locomotion through the tumor stroma, as a prerequisite for establishing contact with target cells, have not been analyzed. These shortcomings limit the rational design of immunotherapeutic strategies that aim to overcome tumor-immune evasion. We have used two-photon microscopy to determine, in a dynamic manner, the requirements leading to tumor regression by TILs. Key observations were that TILs migrated randomly throughout the tumor microenvironment and that, in the absence of cognate antigen, they were incapable of sustaining active migration. Furthermore, TILs in regressing tumors formed long-lasting (>or=30 min), cognate antigen-dependent contacts with tumor cells. Finally, TILs physically interacted with macrophages, suggesting tumor antigen cross-presentation by these cells. Our results demonstrate that recognition of cognate antigen within tumors is a critical determinant of optimal TIL migration and target cell interactions, and argue against TIL guidance by long-range chemokine gradients. PMID- 17116736 TI - Intraluminal crawling of neutrophils to emigration sites: a molecularly distinct process from adhesion in the recruitment cascade. AB - The prevailing view is that the beta2-integrins Mac-1 (alphaMbeta2, CD11b/CD18) and LFA-1 (alphaLbeta2, CD11a/CD18) serve similar biological functions, namely adhesion, in the leukocyte recruitment cascade. Using real-time and time-lapse intravital video-microscopy and confocal microscopy within inflamed microvessels, we systematically evaluated the function of Mac-1 and LFA-1 in the recruitment paradigm. The chemokine macrophage inflammatory protein-2 induced equivalent amounts of adhesion in wild-type and Mac-1-/- mice but very little adhesion in LFA-1-/- mice. Time-lapse video-microscopy within the postcapillary venules revealed that immediately upon adhesion, there is significant intraluminal crawling of all neutrophils to distant emigration sites in wild-type mice. In dramatic contrast, very few Mac-1-/- neutrophils crawled with a 10-fold decrease in displacement and a 95% reduction in velocity. Therefore, Mac-1-/- neutrophils initiated transmigration closer to the initial site of adhesion, which in turn led to delayed transmigration due to movement through nonoptimal emigration sites. Interestingly, the few LFA-1-/- cells that did adhere crawled similarly to wild-type neutrophils. Intercellular adhesion molecule-1 but not intercellular adhesion molecule-2 mediated the Mac-1-dependent crawling. These in vivo results clearly delineate two fundamentally different molecular mechanisms for LFA-1 and Mac-1 in vivo, i.e., LFA-1-dependent adhesion followed by Mac-1-dependent crawling, and both steps ultimately contribute to efficient emigration out of the vasculature. PMID- 17116737 TI - Insulin-induced remission in new-onset NOD mice is maintained by the PD-1-PD-L1 pathway. AB - The past decade has seen a significant increase in the number of potentially tolerogenic therapies for treatment of new-onset diabetes. However, most treatments are antigen nonspecific, and the mechanism for the maintenance of long term tolerance remains unclear. In this study, we developed an antigen-specific therapy, insulin-coupled antigen-presenting cells, to treat diabetes in nonobese diabetic mice after disease onset. Using this approach, we demonstrate disease remission, inhibition of pathogenic T cell proliferation, decreased cytokine production, and induction of anergy. Moreover, we show that robust long-term tolerance depends on the programmed death 1 (PD-1)-programmed death ligand (PD L)1 pathway, not the distinct cytotoxic T lymphocyte-associated antigen 4 pathway. Anti-PD-1 and anti-PD-L1, but not anti-PD-L2, reversed tolerance weeks after tolerogenic therapy by promoting antigen-specific T cell proliferation and inflammatory cytokine production directly in infiltrated tissues. PD-1-PD-L1 blockade did not limit T regulatory cell activity, suggesting direct effects on pathogenic T cells. Finally, we describe a critical role for PD-1-PD-L1 in another powerful immunotherapy model using anti-CD3, suggesting that PD-1-PD-L1 interactions form part of a common pathway to selectively maintain tolerance within the target tissues. PMID- 17116738 TI - Spontaneous autoimmunity prevented by thymic expression of a single self-antigen. AB - The expression of self-antigen in the thymus is believed to be responsible for the deletion of autoreactive T lymphocytes, a critical process in the maintenance of unresponsiveness to self. The Autoimmune regulator (Aire) gene, which is defective in the disorder autoimmune polyglandular syndrome type 1, has been shown to promote the thymic expression of self-antigens. A clear link, however, between specific thymic self-antigens and a single autoimmune phenotype in this model has been lacking. We show that autoimmune eye disease in aire-deficient mice develops as a result of loss of thymic expression of a single eye antigen, interphotoreceptor retinoid-binding protein (IRBP). In addition, lack of IRBP expression solely in the thymus, even in the presence of aire expression, is sufficient to trigger spontaneous eye-specific autoimmunity. These results suggest that failure of thymic expression of selective single self-antigens can be sufficient to cause organ-specific autoimmune disease, even in otherwise self tolerant individuals. PMID- 17116739 TI - A peroxisomal acyltransferase in mouse identifies a novel pathway for taurine conjugation of fatty acids. AB - A wide variety of endogenous carboxylic acids and xenobiotics are conjugated with amino acids, before excretion in urine or bile. The conjugation of carboxylic acids and bile acids with taurine and glycine has been widely characterized, and de novo synthesized bile acids are conjugated to either glycine or taurine in peroxisomes. Peroxisomes are also involved in the oxidation of several other lipid molecules, such as very long chain acyl-CoAs, branched chain acyl-CoAs, and prostaglandins. In this study, we have now identified a novel peroxisomal enzyme called acyl-coenzyme A:amino acid N-acyltransferase (ACNAT1). Recombinantly expressed ACNAT1 acts as an acyltransferase that efficiently conjugates very long chain and long-chain fatty acids to taurine. The enzyme shows no conjugating activity with glycine, showing that it is a specific taurine conjugator. Acnat1 is mainly expressed in liver and kidney, and the gene is localized in a gene cluster, together with two further acyltransferases, one of which conjugates bile acids to glycine and taurine. In conclusion, these data describe ACNAT1 as a new acyltransferase, involved in taurine conjugation of fatty acids in peroxisomes, identifying a novel pathway for production of N-acyltaurines as signaling molecules or for excretion of fatty acids. PMID- 17116740 TI - Progestins overcome inhibition of platelet aggregation by endothelial cells by down-regulating endothelial NO synthase via glucocorticoid receptors. AB - Hormone replacement therapy with estroprogestin preparations is associated with an increased risk of venous and arterial thromboembolic events in postmenopausal women. This study examined whether progestins affect the formation of NO in endothelial cells, and, if so, to determine the underlying mechanism. Experiments were performed with human umbilical vein endothelial cells. Endothelial nitric oxide synthase (eNOS) expression was assessed by real-time polymerase chain reaction (PCR) and Western blot analysis, NO formation by electron spin resonance spectroscopy, nuclear translocation of the glucocorticoid receptor by immunofluorescence microscopy, and platelet aggregation by an aggregometer. Medroxyprogesterone acetate (MPA) and progesterone markedly decreased the eNOS mRNA and protein levels, whereas levonorgestrel and nomegestrol acetate had only small effects. This effect was associated with a decreased NO formation leading to a reduced ability of endothelial cells to prevent platelet aggregation and was prevented by knockdown of the glucocorticoid receptor using siRNA. MPA and progesterone, but not levonorgestrel and nomegestrol acetate, caused nuclear translocation of the glucocorticoid receptor. The present findings indicate that certain progestins, including MPA, reduce the antiaggregatory effect of endothelial cells by decreasing the expression of eNOS and the formation of NO in endothelial cells, an effect that is mediated via activation of glucocorticoid receptors. PMID- 17116741 TI - BMP-7 functions as a novel hormone to facilitate liver regeneration. AB - Bone morphogenic protein-7 (BMP-7) is a key protein involved in liver organogenesis and development. The physiological circulating concentration of BMP 7 is between 100 and 300 pg/ml. BMP-7 expression is absent in the liver, but the receptors for BMP-7 are present on adult hepatocytes. Therefore, we hypothesized that BMP-7 might function as an endogenous regulator of adult hepatocyte proliferation and liver homeostasis. Here, we demonstrate that neutralization of circulating endogenous BMP-7 results in significantly impaired regeneration of the liver after partial hepatectomy. Therapeutic administration of recombinant human BMP-7 (rhBMP-7) significantly enhances liver regeneration associated with accelerated improvement of liver function. Collectively, our results argue for the role of BMP-7 as a kidney- and bone-produced endogenous regulator of hepatocyte health. PMID- 17116742 TI - Cell membranes and liposomes dissociate C-reactive protein (CRP) to form a new, biologically active structural intermediate: mCRP(m). AB - Emerging evidence indicates that C-reactive protein (CRP) has at least two conformationally distinct isoforms, i.e., pentameric CRP (pCRP) and monomeric CRP (mCRP or CRP subunit). Both CRP isoforms are proposed to play roles in inflammation and may participate in the pathogenesis of cardiovascular disease. However, the origin of mCRP in situ and the interplay between the two CRP isoforms under physiological/pathological circumstances remain elusive. Herein, by probing conformational alteration, neoepitope expression, and direct visualization using electron-microscopy, we have shown that calcium-dependent binding of pCRP to membranes, including liposomes and cell membranes, led to a rapid but partial structural change, producing molecules that express CRP subunit antigenicity but with retained native pentameric conformation. This hybrid molecule is herein termed mCRP(m). The formation of mCRP(m) was associated with significantly enhanced complement fixation. mCRP(m) can further detach from membrane to form the well-recognized mCRP isoform converted in solution (mCRP(s)) and exert potent stimulatory effects on endothelial cells. The membrane-induced pCRP dissociation not only provides a physiologically relevant scenario for mCRP formation but may represent an important mechanism for regulating CRP function. PMID- 17116743 TI - Olfactory uptake of manganese requires DMT1 and is enhanced by anemia. AB - Manganese, an essential nutrient, can also elicit toxicity in the central nervous system (CNS). The route of exposure strongly influences the potential neurotoxicity of manganese-containing compounds. Recent studies suggest that inhaled manganese can enter the rat brain through the olfactory system, but little is known about the molecular factors involved. Divalent metal transporter 1 (DMT1) is the major transporter responsible for intestinal iron absorption and its expression is regulated by body iron status. To examine the potential role of this transporter in uptake of inhaled manganese, we studied the Belgrade rat, since these animals display significant defects in both iron and manganese metabolism due to a glycine-to-arginine substitution (G185R) in their DMT1 gene product. Absorption of intranasally instilled 54Mn was significantly reduced in Belgrade rats and was enhanced in iron-deficient rats compared to iron-sufficient controls. Immunohistochemical experiments revealed that DMT1 was localized to both the lumen microvilli and end feet of the sustentacular cells of the olfactory epithelium. Importantly, we found that DMT1 protein levels were increased in anemic rats. The apparent function of DMT1 in olfactory manganese absorption suggests that the neurotoxicity of the metal can be modified by iron status due to the iron-responsive regulation of the transporter. PMID- 17116744 TI - Rapid disuse and denervation atrophy involve transcriptional changes similar to those of muscle wasting during systemic diseases. AB - We previously identified a common set of genes, termed atrogenes, whose expression is coordinately induced or suppressed in muscle during systemic wasting states (fasting, cancer cachexia, renal failure, diabetes). To determine whether this transcriptional program also functions during atrophy resulting from loss of contractile activity and whether atrogene expression correlates with the rate of muscle weight loss, we used cDNA microarrays and RT-polymerase chain reaction to analyze changes in mRNA from rat gastrocnemius during disuse atrophy induced by denervation or spinal cord isolation. Three days after Den or SI, the rate of muscle weight loss was greatest, and 78% of the atrogenes identified during systemic catabolic states were induced or repressed. Of particular interest were the large inductions of key ubiquitin ligases, atrogin-1 (35- to 44 fold) and MuRF1 (12- to 22-fold), and the suppression of PGC-1alpha and PGC-1beta coactivators (15-fold). When atrophy slowed (day 14), the expression of 92% of these atrogenes returned toward basal levels. At 28 days, the atrophy-inducing transcription factor, FoxO1, was still induced and may be important in maintaining the "atrophied" state. Thus, 1) the atrophy associated with systemic catabolic states and following disuse involves similar transcriptional adaptations; and 2) disuse atrophy proceeds through multiple phases corresponding to rapidly atrophying and atrophied muscles that involve distinct transcriptional patterns. PMID- 17116745 TI - Hydrogen sulfide-induces DNA damage and changes in apoptotic gene expression in human lung fibroblast cells. AB - Hydrogen sulfide (H2S) has been shown previously to exert proapoptotic activity. However, the mechanism(s) by which H2S affects cell growth and function have not been addressed adequately. In this study, cultured human lung fibroblasts were treated with the H2S donor NaHS (10-75 microM; 12-48 h). NaHS caused a concentration-dependent increase in micronuclei formation (indicating DNA damage) and cell cycle arrest (G1 phase). NaHS increased expression of ku 70 and ku 80 but did not affect the expression of other DNA repair proteins such as proliferating cell nuclear antigen (PCNA) or replication protein A (rNase protection assay). NaHS treatment also resulted in stabilization of p53 coupled with induction of downstream proteins such as p21, Bax, and cytochrome c, as well as translocation of Bax from the cytosol to the mitochondria and release of cytochrome c from mitochondria. NaHS did not up-regulate cell levels of the antiapoptotic protein, Bcl-2. We propose that the genotoxic action of H2S propels the cell toward apoptotic death triggered initially by stabilization of p53 and subsequently involving a cascade of downstream products. These results are of significance as they uncover a hitherto unknown and very fundamental role for H2S in determining cell fate. PMID- 17116746 TI - Methylation of DNA polymerase beta by protein arginine methyltransferase 1 regulates its binding to proliferating cell nuclear antigen. AB - DNA polymerase beta (pol beta) is a key player in DNA base excision repair (BER). Here, we describe the complex formation of pol beta with the protein arginine methyltransferase 1 (PRMT1). PRMT1 specifically methylated pol beta in vitro and in vivo. Arginine 137 was identified in pol beta as an important target for methylation by PRMT1. Neither the polymerase nor the dRP-lyase activities of pol beta were affected by PRMT1 methylation. However, this modification abolished the interaction of pol beta with proliferating cell nuclear antigen (PCNA). Together, our results provide evidence that PRMT1 methylation of pol beta might play a regulatory role in BER by preventing the involvement of pol beta in PCNA dependent DNA metabolic events. PMID- 17116747 TI - A pivotal role of matrix metalloproteinase-3 activity in dopaminergic neuronal degeneration via microglial activation. AB - Recent studies have demonstrated that activated microglia play an important role in dopamine (DA) neuronal degeneration in Parkinson disease (PD) by generating NADPH-oxidase (NADPHO)-derived superoxide. However, the molecular mechanisms that underlie microglial activation in DA cell death are still disputed. We report here that matrix metalloproteinase-3 (MMP-3) was newly induced and activated in stressed DA cells, and the active form of MMP-3 (actMMP-3) was released into the medium. The released actMMP-3, as well as catalytically active recombinant MMP-3 (cMMP-3) led to microglial activation and superoxide generation in microglia and enhanced DA cell death. cMMP-3 caused DA cell death in mesencephalic neuron-glia mixed culture of wild-type (WT) mice, but this was attenuated in the culture of NADPHO subunit null mice (gp91(phox-/-)), suggesting that NADPHO mediated the cMMP-3-induced microglial production of superoxide and DA cell death. Furthermore, in the N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-injected animal model of PD, nigrostriatal DA neuronal degeneration, microglial activation, and superoxide generation were largely attenuated in MMP-3-/- mice. These results indicate that actMMP-3 released from stressed DA neurons is responsible for microglial activation and generation of NADPHO-derived superoxide and eventually enhances nigrostriatal DA neuronal degeneration. Our results could lead to a novel therapeutic approach to PD. PMID- 17116748 TI - High levels of Notch signaling down-regulate Numb and Numblike. AB - Inhibition of Notch signaling by Numb is critical for many cell fate decisions. In this study, we demonstrate a more complex relationship between Notch and the two vertebrate Numb homologues Numb and Numblike. Although Numb and Numblike at low levels of Notch signaling negatively regulated Notch, high levels of Notch signaling conversely led to a reduction of Numb and Numblike protein levels in cultured cells and in the developing chick central nervous system. The Notch intracellular domain but not the canonical Notch downstream proteins Hes 1 and Hey 1 caused a reduction of Numb and Numblike. The Notch-mediated reduction of Numblike required the PEST domain in the Numblike protein and was blocked by the proteasome inhibitor MG132. Collectively, these observations reveal a reciprocal negative regulation between Notch and Numb/Numblike, which may be of relevance for stabilizing asymmetric cell fate switches and for tumor development. PMID- 17116749 TI - Comparative proteomics of clathrin-coated vesicles. AB - Clathrin-coated vesicles (CCVs) facilitate the transport of cargo between the trans-Golgi network, endosomes, and the plasma membrane. This study presents the first comparative proteomics investigation of CCVs. A CCV-enriched fraction was isolated from HeLa cells and a "mock CCV" fraction from clathrin-depleted cells. We used a combination of 2D difference gel electrophoresis and isobaric tags for relative and absolute quantification (iTRAQ) in conjunction with mass spectrometry to analyze and compare the two fractions. In total, 63 bona fide CCV proteins were identified, including 28 proteins whose association with CCVs had not previously been established. These include numerous post-Golgi SNAREs; subunits of the AP-3, retromer, and BLOC-1 complexes; lysosomal enzymes; CHC22; and five novel proteins of unknown function. The strategy outlined in this paper should be widely applicable as a means of distinguishing genuine organelle components from contaminants. PMID- 17116750 TI - Simple kinetic relationships and nonspecific competition govern nuclear import rates in vivo. AB - Many cargoes destined for nuclear import carry nuclear localization signals that are recognized by karyopherins (Kaps). We present methods to quantitate import rates and measure Kap and cargo concentrations in single yeast cells in vivo, providing new insights into import kinetics. By systematically manipulating the amounts, types, and affinities of Kaps and cargos, we show that import rates in vivo are simply governed by the concentrations of Kaps and their cargo and the affinity between them. These rates fit to a straightforward pump-leak model for the import process. Unexpectedly, we deduced that the main limiting factor for import is the poor ability of Kaps and cargos to find each other in the cytoplasm in a background of overwhelming nonspecific competition, rather than other more obvious candidates such as the nuclear pore complex and Ran. It is likely that most of every import round is taken up by Kaps and nuclear localization signals sampling other cytoplasmic proteins as they locate each other in the cytoplasm. PMID- 17116751 TI - IKKbeta programs to turn on the GADD45alpha-MKK4-JNK apoptotic cascade specifically via p50 NF-kappaB in arsenite response. AB - Cross talk between NF-kappaB and c-Jun N-terminal kinases (JNKs) has been implicated in the cell life and death decision under various stresses. Functional suppression of JNK activation by NF-kappaB has recently been proposed as a key cellular survival mechanism and contributes to cancer cells escaping from apoptosis. We provide a novel scenario of the proapoptotic role of IkappaB kinase beta (IKKbeta)-NF-kappaB, which can act as the activator of the JNK pathway through the induction of GADD45alpha for triggering MKK4/JNK activation, in response to the stimulation of arsenite, a cancer therapeutic reagent. This effect of IKKbeta-NF-kappaB is dependent on p50 but not the p65/relA NF-kappaB subunit, which can increase the stability of GADD45alpha protein through suppressing its ubiquitination and proteasome-dependent degradation. IKKbeta-NF kappaB can therefore either activate or suppress the JNK cascade and consequently mediate pro- or antiapoptotic effects, depending on the manner of its induction. Furthermore, the NF-kappaB p50 subunit can exert a novel regulatory function on protein modification independent of the classical NF-kappaB transcriptional activity. PMID- 17116752 TI - The major human and mouse granzymes are structurally and functionally divergent. AB - Approximately 2% of mammalian genes encode proteases. Comparative genomics reveals that those involved in immunity and reproduction show the most interspecies diversity and evidence of positive selection during evolution. This is particularly true of granzymes, the cytotoxic proteases of natural killer cells and CD8+ T cells. There are 5 granzyme genes in humans and 10 in mice, and it is suggested that granzymes evolve to meet species-specific immune challenge through gene duplication and more subtle alterations to substrate specificity. We show that mouse and human granzyme B have distinct structural and functional characteristics. Specifically, mouse granzyme B is 30 times less cytotoxic than human granzyme B and does not require Bid for killing but regains cytotoxicity on engineering of its active site cleft. We also show that mouse granzyme A is considerably more cytotoxic than human granzyme A. These results demonstrate that even "orthologous" granzymes have species-specific functions, having evolved in distinct environments that pose different challenges. PMID- 17116753 TI - Transport of LAPTM5 to lysosomes requires association with the ubiquitin ligase Nedd4, but not LAPTM5 ubiquitination. AB - LAPTM5 is a lysosomal transmembrane protein expressed in immune cells. We show that LAPTM5 binds the ubiquitin-ligase Nedd4 and GGA3 to promote LAPTM5 sorting from the Golgi to the lysosome, an event that is independent of LAPTM5 ubiquitination. LAPTM5 contains three PY motifs (L/PPxY), which bind Nedd4-WW domains, and a ubiquitin-interacting motif (UIM) motif. The Nedd4-LAPTM5 complex recruits ubiquitinated GGA3, which binds the LAPTM5-UIM; this interaction does not require the GGA3-GAT domain. LAPTM5 mutated in its Nedd4-binding sites (PY motifs) or its UIM is retained in the Golgi, as is LAPTM5 expressed in cells in which Nedd4 or GGA3 is knocked-down with RNAi. However, ubiquitination-impaired LAPTM5 can still traffic to the lysosome, suggesting that Nedd4 binding to LAPTM5, not LAPTM5 ubiquitination, is required for targeting. Interestingly, Nedd4 is also able to ubiquitinate GGA3. These results demonstrate a novel mechanism by which the ubiquitin-ligase Nedd4, via interactions with GGA3 and cargo (LAPTM5), regulates cargo trafficking to the lysosome without requiring cargo ubiquitination. PMID- 17116755 TI - Intrarenal dopamine D1-like receptor stimulation induces natriuresis via an angiotensin type-2 receptor mechanism. AB - We explored the effects of direct renal interstitial stimulation of dopamine D(1) like receptors with fenoldopam, a selective D(1)-like receptor agonist, on renal sodium excretion and angiotensin type-2 (AT(2)) receptor expression and cellular distribution in rats on a high-sodium intake. In contrast to vehicle-infused rats, sodium excretion increased in fenoldopam-infused rats during each of three 1-hour experimental periods (<0.001). Blood pressure was unaffected by vehicle or fenoldopam. In plasma membranes of renal cortical cells, fenoldopam increased D(1) receptor expression by 38% (P<0.05) and AT(2) receptor expression by 69% (P<0.01). In plasma membranes of renal proximal tubule cells, fenoldopam increased AT(2) receptor expression by 108% (P<0.01). In outer apical membranes of proximal tubule cells, fenoldopam increased AT(2) receptor expression by 59% (P<0.01). No significant change in total AT(2) receptor protein expression was detectable in response to fenoldopam. Fenoldopam-induced natriuresis was abolished when either PD-123319, a specific AT(2) receptor antagonist, or SCH 23390, a potent D(1)-like receptor antagonist, was coinfused with F (P<0.001). In summary, direct renal D(1)-like receptor activation increased urinary sodium excretion and the plasma membrane expression of AT(2) receptors in renal cortical and proximal tubule cells. D(1)-like receptor-induced natriuresis was abolished by intrarenal AT(2) receptor inhibition. These findings suggest that dopaminergic regulation of sodium excretion involves recruitment of AT(2) receptors to the outer plasma membranes of renal proximal tubule cells and that dopamine-induced natriuresis requires AT(2) receptor activation. PMID- 17116754 TI - Lipid defect underlies selective skin barrier impairment of an epidermal-specific deletion of Gata-3. AB - Skin lies at the interface between the complex physiology of the body and the external environment. This essential epidermal barrier, composed of cornified proteins encased in lipids, prevents both water loss and entry of infectious or toxic substances. We uncover that the transcription factor GATA-3 is required to establish the epidermal barrier and survive in the ex utero environment. Analysis of Gata-3 mutant transcriptional profiles at three critical developmental stages identifies a specific defect in lipid biosynthesis and a delay in differentiation. Genomic analysis identifies highly conserved GATA-3 binding sites bound in vivo by GATA-3 in the first intron of the lipid acyltransferase gene AGPAT5. Skin from both Gata-3-/- and previously characterized barrier deficient Kruppel-like factor 4-/- newborns up-regulate antimicrobial peptides, effectors of innate immunity. Comparison of these animal models illustrates how impairment of the skin barrier by two genetically distinct mechanisms leads to innate immune responses, as observed in the common human skin disorders psoriasis and atopic dermatitis. PMID- 17116756 TI - Angiotensin-(1-7) through receptor Mas mediates endothelial nitric oxide synthase activation via Akt-dependent pathways. AB - Angiotensin-(1-7) [Ang-(1-7)] causes endothelial-dependent vasodilation mediated, in part, by NO release. However, the molecular mechanisms involved in endothelial NO synthase (eNOS) activation by Ang-(1-7) remain unknown. Using Chinese hamster ovary cells stably transfected with Mas cDNA (Chinese hamster ovary-Mas), we evaluated the underlying mechanisms related to receptor Mas-mediated posttranslational eNOS activation and NO release. We further examined the Ang-(1 7) profile of eNOS activation in human aortic endothelial cells, which constitutively express the Mas receptor. Chinese hamster ovary-Mas cells and human aortic endothelial cell were stimulated with Ang-(1-7; 10(-7) mol/L; 1 to 30 minutes) in the absence or presence of A-779 (10(-6) mol/L). Additional experiments were performed in the presence of the phosphatidylinositol 3-kinase inhibitor wortmannin (10(-6) mol/L). Changes in eNOS (at Ser1177/Thr495 residues) and Akt phosphorylation were evaluated by Western blotting. NO release was measured using both the fluorochrome 2,3-diaminonaphthalene and an NO analyzer. Ang-(1-7) significantly stimulated eNOS activation (reciprocal phosphorylation/dephosphorylation at Ser1177/Thr495) and induced a sustained Akt phosphorylation (P<0.05). Concomitantly, a significant increase in NO release was observed (2-fold increase in relation to control). These effects were blocked by A-779. Wortmannin suppressed eNOS activation in both Chinese hamster ovary-Mas and human aortic endothelial cells. Our findings demonstrate that Ang-(1-7), through Mas, stimulates eNOS activation and NO production via Akt-dependent pathways. These novel data highlight the importance of the Ang-(1-7)/Mas axis as a putative regulator of endothelial function. PMID- 17116757 TI - Adrenergic overdrive as the link among hypertension, obesity, and impaired thermogenesis: lights and shadows. PMID- 17116758 TI - Autonomic contribution to blood pressure and metabolism in obesity. AB - Obesity is associated with alterations in the autonomic nervous system that may contribute to the increase in blood pressure and resting energy expenditure present in this condition. To test this hypothesis, we induced autonomic withdrawal with the ganglionic blocker trimethaphan in 10 lean (32+/-3 years) and 10 obese (35+/-3 years) subjects. Systolic blood pressure fell more in obese compared with lean subjects (-17+/-3 versus -11+/-1 mm Hg; P=0.019) because of a greater decrease in total peripheral resistance (-310+/-41 versus 33+/-78 dynes/sec/cm(-5); P=0.002). In contrast, resting energy expenditure decreased less in obese than in lean subjects, (-26+/-21 versus -86+/-15 kcal per day adjusted by fat-free mass; P=0.035). We confirmed that the autonomic contribution to blood pressure was greater in obesity after including additional subjects with a wider range of blood pressures. Systolic blood pressure decreased -28+/-4 mm Hg (95% CI: -38 to -18.0; n=8) in obese hypertensive subjects compared with lean ( 9+/-1 mm Hg; 95% CI: -11 to -6; n=22) or obese normotensive subjects (-14+/-2 mm Hg; 95% CI: -18 to -10; n=20). After removal of autonomic influences, systolic blood pressure remained higher in obese hypertensive subjects (109+/-3 versus 98+/-2 mm Hg in lean and 103+/-2 mm Hg in obese normotensive subjects; P=0.004) suggesting a role for additional factors in obesity-associated hypertension. In conclusion, sympathetic activation induced by obesity is an important determinant to the blood pressure elevation associated with this condition but is not effective in increasing resting energy expenditure. These results suggest that the sympathetic nervous system could be targeted in the treatment of obesity associated hypertension. PMID- 17116759 TI - Multilocus analyses of Renin-Angiotensin-aldosterone system gene variants on blood pressure at rest and during behavioral stress in young normotensive subjects. AB - The renin-angiotensin-aldosterone system (RAAS) is a proteolytic cascade that regulates and maintains blood pressure (BP). This study aimed to explore the interactive and integrative effects of multiple RAAS polymorphisms on BP at rest and during behavioral stress in a normotensive population. A total of 920 young white and black twins (age: 12 to 30 years; 45% blacks) was subjected to three 10 minute stress tasks. Thirteen potential functional polymorphisms from 4 major RAAS genes were genotyped. We performed multilocus prediction allowing for genetic modification effects (gene-gene, gene-gender, gene-ethnicity, and gene body mass index) using Multivariate Adaptive Regression Splines and generalized estimating equations. Single polymorphism analyses showed modest effects of M235T (angiotensinogen) and A-239T (angiotensin I-converting enzyme; P value range: 0.005 to 0.036), accounting for approximately 1% of the total variance of systolic BP at rest and during stress. Compared with this, the best multilocus models revealed multiple independent genetic modification effects (gene-gene, gene-gender, and gene-body mass index; P value range: 0.003 to 0.009), accounting for 2.5% and 7.3% of the total variance for systolic BP levels at rest and during stress, respectively. Our data support the hypothesis that multiple RAAS genetic modifications account for BP variation. We conclude that the RAAS genetic modifications may contribute more to the dynamic BP regulation in response to behavioral stress compared with the static BP value. In addition, we reported a gene-gene interaction between M235T (angiotensinogen) and A1159G (angiotensin I converting enzyme) on stress systolic BP levels. We proposed a viable approach to test for the multiple genetic contributions to BP and hypertension. PMID- 17116760 TI - Maternal fetal/placental interactions and abnormal pregnancy outcomes. PMID- 17116761 TI - Endothelial dysfunction: a link among preeclampsia, recurrent pregnancy loss, and future cardiovascular events? AB - We tested the hypothesis that endothelial dysfunction could cause placentation related defects, persist after the complicated pregnancy, and probably cause cardiovascular disease later in life. Brachial arterial reactivity and factors related to endothelial dysfunction, such as circulating cholesterol, uric acid, nitrites, l-arginine, asymmetrical dimethylarginine, vascular endothelial growth factor, and soluble vascular endothelial growth factor receptor-1, in women with previous healthy pregnancies (n=22), patients with severe preeclampsia (n=25), or patients with recurrent pregnancy loss (n=29), at day 10 of the luteal phase of an ovulatory cycle an average of 11 to 27 months after pregnancy were evaluated. Both groups with placentation defects had a significant decrease in endothelium dependent dilatation, a higher rate of endothelial dysfunction, lower serum nitrites, and higher cholesterol as compared with control subjects; subjects with previous preeclampsia additionally had higher normal blood pressures and a greater parental prevalence of cardiovascular disease. Patients with recurrent pregnancy loss also demonstrated a significantly lower endothelium-independent vasodilatation. A trend to an inverse correlation was found between serum cholesterol serum and endothelial-mediated vasodilatation in the whole study population. Uric acid, l-arginine, asymmetrical dimethylarginine, vascular endothelial growth factor, and soluble vascular endothelial growth factor receptor-1 were similar in all of the groups. We postulate that endothelial dysfunction may represent a link between preeclampsia and increased cardiovascular disease latter in life and propose that women with unexplained recurrent miscarriages are also at increased cardiovascular risk. The identification and correction of endothelial dysfunction detected during the reproductive stage on obstetric outcome and on cardiovascular diseases needs to be elucidated. PMID- 17116762 TI - Inositol phosphoglycan P-type in preeclampsia: a novel marker? AB - A state of insulin resistance has been demonstrated in active preeclampsia, and women with clinical evidence of insulin resistance are at higher risk to develop this syndrome during pregnancy. Recently, inositol phosphoglycan P-type, a putative second messenger of insulin action, has been implicated in the pathophysiology of preeclampsia and is increased in the placenta, amniotic fluid, and maternal urine of preeclamptic women compared with normal pregnant women. We report here a case-control study to assess the potential of urinary levels of inositol phosphoglycan P-type as a screening test for preeclampsia. Twenty-seven preeclamptic women and 47 healthy pregnant women were recruited. A polyclonal antibody-based ELISA was developed to detect levels of inositol phosphoglycan P type in urine. Its content in urinary specimens was found to be 30-fold higher in preeclamptic subjects than control subjects (329.1+/-21.8 versus 9.2+/-1.5; P<0.001), with a higher level in all of the preeclamptic cases. For 6 women who developed preeclampsia, >1 gestational date sample of urine was available, and retrospective analysis showed a significant time-related increase of the urinary level of inositol phosphoglycan P-type or = 100,000 dollars) decreased progressively with increasing education or income levels (all P<0.001), whereas an opposite pattern was observed for high-density lipoprotein (P<0.001). Overall, in age-adjusted Cox proportional hazards models, the relative risk of incident CVD events decreased with increasing education (1.0, 0.7, 0.5, 0.4, and 0.5; P for trend <0.001) and income (1.0, 1.0, 0.9, 0.7, 0.6, and 0.4; P for trend <0.001) categories. In multivariate models that assessed the impact of traditional and novel CVD risk factors on the relationship between education/income and CVD events, the relative hazard of incident CVD associated with a 1-category-higher level of education changed from 0.79 in age- and race-adjusted analysis to 0.89 in fully adjusted analysis. The 11% lower risk per 1 category of education remained significant (P for trend=0.006), suggesting that controlling for both novel and traditional risk factors could not explain the protective effect of education. A similar analysis for income revealed that its relationship with CVD events was explained largely by these noted risk factors. CONCLUSIONS: In this prospective analysis, we observed a decrease in incident CVD events with increasing levels of education and income. In contrast to the relationship between income and CVD events, the relationship of CVD events with education was explained only partially by traditional and novel risk factors for CVD. PMID- 17116765 TI - Pathogen-sensing plasmacytoid dendritic cells stimulate cytotoxic T-cell function in the atherosclerotic plaque through interferon-alpha. AB - BACKGROUND: Unstable atherosclerotic plaque is characterized by an infiltrate of inflammatory cells. Both macrophages and T cells have been implicated in mediating the tissue injury leading to plaque rupture; however, signals regulating their activation remain unidentified. Infectious episodes have been suspected to render plaques vulnerable to rupture. We therefore explored whether plasmacytoid dendritic cells (pDCs) that specialize in sensing bacterial and viral products can regulate effector functions of plaque-residing T cells and thus connect host infection and plaque instability. METHODS AND RESULTS: pDCs were identified in 53% of carotid atheromas (n=30) in which they localized to the shoulder region and produced the potent immunoregulatory cytokine interferon (INF)-alpha. IFN-alpha transcript concentrations in atheroma tissues correlated strongly with plaque instability (P<0.0001). Plaque-residing pDCs responded to pathogen-derived motifs, CpG-containing oligodeoxynucleotides binding to toll like receptor 9, with enhanced IFN-alpha transcription (P=0.03) and secretion (P=0.007). IFN-alpha emerged as a potent regulator of T-cell function, even in the absence of antigen recognition. Specifically, IFN-alpha induced a 10-fold increase of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on the surface of CD4 T cells (P<0.0001) and enabled them to effectively kill vascular smooth muscle cells (P=0.0003). CONCLUSIONS: pDCs in atherosclerotic plaque sense microbial motifs and amplify cytolytic T-cell functions, thus providing a link between host-infectious episodes and acute immune-mediated complications of atherosclerosis. PMID- 17116766 TI - A once-daily, oral, direct Factor Xa inhibitor, rivaroxaban (BAY 59-7939), for thromboprophylaxis after total hip replacement. AB - BACKGROUND: Rivaroxaban (BAY 59-7939)--an oral, direct Factor Xa inhibitor--could be an alternative to heparins and warfarin for the prevention and treatment of thromboembolic disorders. METHODS AND RESULTS: This randomized, double-blind, double-dummy, active-comparator-controlled, multinational, dose-ranging study assessed the efficacy and safety of once-daily rivaroxaban relative to enoxaparin for prevention of venous thromboembolism in patients undergoing elective total hip replacement. Patients (n=873) were randomized to once-daily oral rivaroxaban doses of 5, 10, 20, 30, or 40 mg (initiated 6 to 8 hours after surgery) or a once daily subcutaneous enoxaparin dose of 40 mg (given the evening before and > or = 6 hours after surgery). Study drugs were continued for an additional 5 to 9 days; mandatory bilateral venography was performed the following day. The primary end point (composite of any deep vein thrombosis, objectively confirmed pulmonary embolism, and all-cause mortality) was observed in 14.9%, 10.6%, 8.5%, 13.5%, 6.4%, and 25.2% of patients receiving 5, 10, 20, 30, and 40 mg rivaroxaban, and 40 mg enoxaparin, respectively (n=618, per-protocol population). No significant dose-response relationship was found for efficacy (P=0.0852). Major postoperative bleeding was observed in 2.3%, 0.7%, 4.3%, 4.9%, 5.1%, and 1.9% of patients receiving 5, 10, 20, 30, and 40 mg rivaroxaban, and 40 mg enoxaparin, respectively (n=845, safety population), representing a significant dose-response relationship (P=0.0391). CONCLUSIONS: Rivaroxaban showed efficacy and safety similar to enoxaparin for thromboprophylaxis after total hip replacement, with the convenience of once-daily oral dosing and without the need for coagulation monitoring. When both efficacy and safety are considered, these results suggest that 10 mg rivaroxaban once daily should be investigated in phase III studies. PMID- 17116767 TI - Extending the horizon in chronic heart failure: effects of multidisciplinary, home-based intervention relative to usual care. AB - BACKGROUND: The long-term impact of chronic heart failure management programs over the typical life span of affected individuals is unknown. METHODS AND RESULTS: The effects of a nurse-led, multidisciplinary, home-based intervention (HBI) in a typically elderly cohort of patients with chronic heart failure initially randomized to either HBI (n=149) or usual postdischarge care (UC) (n=148) after a short-term hospitalization were studied for up to 10 years of follow-up (minimum 7.5 years of follow-up). Study end points were all-cause mortality, event-free survival (event was defined as death or unplanned hospitalization), recurrent hospital stay, and cost per life-year gained. Median survival in the HBI cohort was almost twice that of UC (40 versus 22 months; P<0.001), with fewer deaths overall (HBI, 77% versus 89%; adjusted relative risk, 0.74; 95% CI, 0.53 to 0.80; P<0.001). HBI was associated with prolonged event free survival (median, 7 versus 4 months; P<0.01). HBI patients had more unplanned readmissions (560 versus 550) but took 7 years to overtake UC; the rates of readmission (2.04+/-3.23 versus 3.66+/-7.62 admissions; P<0.05) and related hospital stay (14.8+/-23.0 versus 28.4+/-53.4 days per patient per year; P<0.05) were significantly lower in the HBI group. HBI was associated with 120 more life-years per 100 patients treated compared with UC (405 versus 285 years) at a cost of 1729 dollars per additional life-year gained when we accounted for healthcare costs including the HBI. CONCLUSIONS: In altering the natural history of chronic heart failure relative to UC (via prolonged survival and reduced frequency of recurrent hospitalization), HBI is a remarkably cost- and time effective strategy over the longer term. PMID- 17116768 TI - Clinical features and outcomes of childhood dilated cardiomyopathy: results from a national population-based study. AB - BACKGROUND: Despite considerable mortality, population-based prognostic factors for childhood dilated cardiomyopathy are lacking. METHODS AND RESULTS: A population-based cohort study was undertaken of all children in Australia who presented with cardiomyopathy at age 0 to 10 years between January 1, 1987, and December 31, 1996. A single cardiologist analyzed all cardiac investigations, and a single pathologist analyzed histopathological material. There were 184 subjects with dilated cardiomyopathy. Positive viral identification or lymphocytic myocarditis was found in 30 (68.2%) of 44 cases with available early histology and 8 of 9 cases presenting with sudden death. Freedom from death or transplantation was 72% (95% CI, 65% to 78%) 1 year after presentation and 63% (95% CI, 55% to 70%) at 5 years. By proportional hazards regression analysis, risk factors for death or transplantation comprised age >5 years at presentation (hazard ratio 5.6, 95% CI, 2.6 to 12.0), familial dilated cardiomyopathy (hazard ratio, 2.9; 95% CI, 1.5 to 5.6), lower initial fractional shortening z score (hazard ratio per z-score unit, 0.75; 95% CI, 0.65 to 0.87), and failure to increase fractional shortening z score during follow-up (hazard ratio per unit increase, 0.68; 95% CI, 0.58 to 0.79). At follow-up, 78 (44.6%) of 175 cases diagnosed during life have no symptoms and are not taking any cardiac medication. CONCLUSIONS: Early mortality is high in childhood dilated cardiomyopathy, but the clinical status of long-term survivors is good. This population-based study identifies children at risk of adverse events. PMID- 17116769 TI - High plasma aldosterone levels on admission are associated with death in patients presenting with acute ST-elevation myocardial infarction. AB - BACKGROUND: Aldosterone, the final mediator of the renin-angiotensin-aldosterone pathway, is at its highest plasma levels at presentation for ST-elevation myocardial infarction (STEMI). Whether aldosterone level at presentation for STEMI is associated with adverse outcome remains unknown. METHODS AND RESULTS: Plasma aldosterone levels were measured at presentation in consecutive patients referred for primary percutaneous coronary intervention for STEMI. We assessed the association between aldosterone levels and in-hospital events and mortality during a 6-month follow-up. Of 356 STEMI patients, 23 and 36 died during the hospital stay and 6-month follow-up period, respectively. Nine other patients survived in-hospital cardiac arrest. High aldosterone levels were associated with an almost stepwise increase in rates of in-hospital death (P=0.01), cardiovascular death (P=0.03), heart failure (P=0.005), ventricular fibrillation (P=0.02), and resuscitated cardiac arrest (P=0.01). After adjustment for age, Killip class, and reperfusion status, compared with patients in the first aldosterone quartile group, those in the highest quartile were at higher risk of death (hazard ratio 3.28, 95% CI 1.09 to 9.89, P=0.035) and death or resuscitated cardiac arrest (hazard ratio 3.74, 95% CI 1.40 to 9.98, P=0.008) during the follow-up. CONCLUSIONS: Plasma aldosterone levels on admission among patients referred for primary percutaneous coronary intervention for STEMI are associated with early and late adverse clinical outcomes, including mortality. The association between high aldosterone levels and late mortality is independent of age, heart failure, and reperfusion status. Such results underline the pivotal role of aldosterone and justify a randomized trial to assess the early administration of aldosterone antagonists in the setting of STEMI. PMID- 17116770 TI - Intracellular calcium and vulnerability to fibrillation and defibrillation in Langendorff-perfused rabbit ventricles. AB - BACKGROUND: The role of intracellular calcium (Ca(i)) in defibrillation and vulnerability is unclear. METHODS AND RESULTS: We simultaneously mapped epicardial membrane potential and Ca(i) during shock on T-wave episodes (n=104) and attempted defibrillation episodes (n=173) in 17 Langendorff-perfused rabbit ventricles. Unsuccessful and type B successful defibrillation shocks were followed by heterogeneous distribution of Ca(i), including regions of low Ca(i) surrounded by elevated Ca(i) ("Ca(i) sinkholes") 31+/-12 ms after shock. The first postshock activation then originated from the Ca(i) sinkhole 53+/-14 ms after the shock. No sinkholes were present in type A successful defibrillation. A Ca(i) sinkhole also was present 39+/-32 ms after a shock on T that induced ventricular fibrillation, followed 22+/-15 ms later by propagated wave fronts that arose from the same site. This wave propagated to form a spiral wave and initiated ventricular fibrillation. Thapsigargin and ryanodine significantly decreased the upper limit of vulnerability and defibrillation threshold. We studied an additional 7 rabbits after left ventricular endocardial cryoablation, resulting in a thin layer of surviving epicardium. Ca(i) sinkholes occurred 31+/ 12 ms after the shock, followed in 19+/-7 ms by first postshock activation in 63 episodes of unsuccessful defibrillation. At the Ca(i) sinkhole, the rise of Ca(i) preceded the rise of epicardial membrane potential in 5 episodes. CONCLUSIONS: There is a heterogeneous postshock distribution of Ca(i). The first postshock activation always occurs from a Ca(i) sinkhole. The Ca(i) prefluorescence at the first postshock early site suggests that reverse excitation-contraction coupling might be responsible for the initiation of postshock activations that lead to ventricular fibrillation. PMID- 17116771 TI - Statins activate AMP-activated protein kinase in vitro and in vivo. AB - BACKGROUND: Statins exert pleiotropic effects on the cardiovascular system, in part through an increase in nitric oxide (NO) bioavailability. AMP-activated protein kinase (AMPK) plays a central role in controlling energy and metabolism homeostasis in various organs. We therefore studied whether statins can activate AMPK, and if so, whether the activated AMPK regulates nitric oxide (NO) production and angiogenesis mediated by endothelial NO synthase, a substrate of AMPK in vascular endothelial cells. METHODS AND RESULTS: Western blotting of protein extracts from human umbilical vein endothelial cells treated with atorvastatin revealed increased phosphorylation of AMPK at Thr-172 in a time- and dose-dependent manner. The AMPK activity, assessed by SAMS assay, was also increased accordingly. The phosphorylation of acetyl-CoA carboxylase at Ser-79 and of endothelial NO synthase at Ser-1177, 2 putative downstream targets of AMPK, was inhibited by an adenovirus that expressed a dominant-negative mutant of AMPK (Ad-AMPK-DN) and compound C, an AMPK antagonist. The positive effects of atorvastatin, including NO production, cGMP accumulation, and in vitro angiogenesis in Matrigel, were all blocked by Ad-AMPK-DN. Mice given atorvastatin through gastric gavage showed increased AMPK, acetyl-CoA carboxylase, and endothelial NO synthase phosphorylation in mouse aorta and myocardium. CONCLUSIONS: Statins can rapidly activate AMPK via increased Thr-172 phosphorylation in vitro and in vivo. Such phosphorylation results in endothelial NO synthase activation, which provides a novel explanation for the pleiotropic effects of statins that benefit the cardiovascular system. PMID- 17116772 TI - Outcome after implantation of a cardioverter-defibrillator in patients with Brugada syndrome: a multicenter study. AB - BACKGROUND: Brugada syndrome is an arrhythmogenic disease characterized by an increased risk of sudden cardiac death (SCD) by ventricular fibrillation. At present, an implantable cardioverter-defibrillator (ICD) is the recommended therapy in high-risk patients. This multicenter study reports the outcome of a large series of patients implanted with an ICD for Brugada syndrome. METHODS AND RESULTS: All patients (n=220, 46+/-12 years, 183 male) with a type 1 Brugada ECG pattern implanted with an ICD in 14 centers between 1993 and 2005 were investigated. ICD indication was based on resuscitated SCD (18 patients, 8%), syncope (88 patients, 40%), or positive electrophysiological study in asymptomatic patients (99 patients, 45%). The remaining 15 patients received an ICD because of a family history of SCD or nonsustained ventricular arrhythmia. During a mean follow-up of 38+/-27 months, no patient died and 18 patients (8%) had appropriate device therapy (10+/-15 shocks/patient, 26+/-33 months after implantation). The complication rate was 28%, including inappropriate shocks, which occurred in 45 patients (20%, 4+/-3 shocks/patient, 21+/-20 months after implantation). The reasons for inappropriate therapy were lead failure (19 patients), T-wave oversensing (10 patients), sinus tachycardia (10 patients), and supraventricular tachycardia (9 patients). Among implantation parameters, high defibrillation threshold, high pacing threshold, and low R-wave amplitude occurred, respectively, in 12%, 27%, and 15% of cases. CONCLUSIONS: In this large Brugada syndrome population, a low incidence of arrhythmic events was found, with an annual event rate of 2.6% during a follow-up of >3 years, in addition to a significant risk of device-related complications (8.9%/year). Inappropriate shocks were 2.5 times more frequent than appropriate ones. PMID- 17116773 TI - Images in cardiovascular medicine. Echocardiographic history of an asymptomatic congenital cardiac tumor: no changes in mass dimensions during a 14-year follow up. PMID- 17116774 TI - Images in cardiovascular medicine. Intraamniotic fetal echocardiography: a new fetal cardiovascular monitoring approach during human fetoscopic surgery. PMID- 17116775 TI - Images in cardiovascular medicine. Esophageal diverticulum illustrated by barium swallow during left atrial catheter ablation for atrial fibrillation. PMID- 17116776 TI - Letter by Karapanayiotides regarding article, "Histological assessment of 526 symptomatic carotid plaques in relation to the nature and timing of ischemic symptoms: the Oxford plaque study". PMID- 17116777 TI - Viewpoint: The safety of drug-eluting stents. PMID- 17116778 TI - Hypertrophic cardiomyopathy: the search for obstruction. PMID- 17116779 TI - Increasing evidence that estrogen is an important modulator of bone marrow mediated cardiac repair after acute infarction. PMID- 17116780 TI - Heart transplantation in patients with diabetes. PMID- 17116781 TI - Variable impact of combining fatal and nonfatal end points in heart failure trials. AB - Randomized clinical trials (RCTs) are a cornerstone of evidence-based medicine. Their design, implementation, and interpretation are sometimes subject to flaws and errors. To achieve statistical significance and economically feasible RCTs, the use of composite end points in heart failure (HF) trials has become more common. We analyzed the incremental value of combining HF hospitalizations with all-cause mortality in trials of chronic HF that enrolled >1000 placebo patients and had a mean follow-up >9 months. We tested the assumption that, compared with mortality, combining HF hospitalization with all-cause death would yield a consistently predictable increase in event rate across HF RCTs. Average placebo arm duration of follow-up was determined, and standardized placebo event rates per 100 patient-years were estimated. Twelve major HF RCTs were included in this analysis. There was a substantial relative increase in the event rate ranging from 64% to 134% when a composite end point was used. This increase was not related to disease severity as described by annual mortality rate. The relative contribution of combining HF hospitalization with all-cause mortality was, however, influenced by the duration of the trial. Combining HF hospitalization with all-cause mortality increases the overall event rate in HF clinical trials; the relative increase varies widely and is unrelated to disease severity. Longer duration trials have a more predictable increase in events than short RCTs. Trial duration must be considered when composite end points are used during the design and interpretation of HF RCTs. PMID- 17116782 TI - Treatment of keloids and hypertrophic scars: a meta-analysis and review of the literature. AB - Management of hypertrophic scars and keloids has advanced from crude, invasive methods such as gross excision and radiation to intralesional or topical agents that act on a cellular level. There is no universally accepted treatment regimen and no evidence-based literature to guide management. Our objectives are to present a list of available treatment regimens, their proposed mechanisms of action, and supporting evidence and to perform a meta-analysis of clinical trials to identify treatments with a better-than-even likelihood of improvement. We conducted a PubMed search through October 2005, identifying clinical studies of various treatments for hypertrophic scars and keloids. We graded the quality of each study, delineated the results into favorable vs nonfavorable, and calculated the statistical significance of the findings. The meta-analysis of 70 treatment series for various clinical measures showed a 70% chance of improvement with treatment; however, the mean amount of improvement to be expected was around 60%. There was no statistically significant difference between treatments. Most treatments for keloidal and hypertrophic scarring offer minimal likelihood of improvement. The magnitude of likely permanent improvement in any sign or symptom may be clinically meaningful but far short of cure. Novel therapies deserve further investigation but remain without proven benefit to date. PMID- 17116783 TI - Quantitative analysis of lateral osteotomies in rhinoplasty. AB - OBJECTIVE: To statistically analyze the long-term results of osteotomy after rhinoplasty. DESIGN: In a consecutive series of 51 patients who underwent reduction rhinoplasty from May 1, 2000, through September 30, 2003, all underwent the same method of bilateral lateral osteotomies, performed by one of us. Twenty patients agreed to participate in this study. The follow-up ranged from 7 to 36 months (mean, 12.6 months). By using preoperative and postoperative photographs, comparisons were made of the change in the dorsal width of the nose (the anterior junction of the nasal bones) and of the ventral width of the nose (where the nasal processes of the maxilla meet the body of the maxilla). RESULTS: There was a significant change (P = .003) in the ventral width of the nose after nasal bone osteotomy. There was no significant change (P = .24) in the dorsal width of the nose. CONCLUSIONS: By using this technique of osteotomy, it is possible to narrow the ventral width of the nose with statistical significance. The dorsal width of the nose is maintained in reduction rhinoplasty. To our knowledge, this is the first attempt to quantify the amount of narrowing achieved after nasal osteotomies. PMID- 17116784 TI - Transcutaneous lower eyelid blepharoplasty with fat excision: a shift-resisting paradigm. AB - OBJECTIVE: To prove through our experience that the use of transcutaneous lower eyelid blepharoplasty results in negligible incidence of unacceptable scar and eyelid malposition and that the overall lower eyelid contour is acceptable. A detailed overview of the lower eyelid anatomy and a discussion of the "aging" eyelid are further discussed. DESIGN: Retrospective, observational study. The study population comprised 50 patients (100 eyes) seen at the McCollough Plastic Surgery Clinic, Gulf Shores, Ala, between 2002 and 2003 (45 women and 5 men), who had undergone transcutaneous lower eyelid blepharoplasty with fat excision. Lower eyelid blepharoplasty was performed by the senior surgeon (E.G.M.), and the surgical technique was identical in all cases. The patients were followed up for a minimum of 6 months and a maximum of 2 years. Patients were selected on the basis of return visits to record the findings, documented by consecutive digital photos. By comparing standard blepharoplasty digital views, the patients were assessed by 3 independent unbiased plastic surgeons. This study was performed in a private practice setting. The main outcome measure was mean score for the presence of unacceptable scarring, the presence of lower eyelid malposition, and the overall appearance of the eyelid after transcutaneous lower eyelid blepharoplasty, as assessed with the Garcia-McCollough Scale for Lower Eyelid Appearance. RESULTS: The 50 patients were retrospectively reviewed and analyzed by a group of 3 unbiased plastic surgeons, and there was negligible evidence of lower eyelid contour abnormality, lower eyelid malposition, or easily visible scars. CONCLUSIONS: Transcutaneous lower eyelid blepharoplasty with fat excision is a time-tested method of correcting the undesirable sequelae of the aging eye. This technique not only is a safe and effective manner to rejuvenate the lower eyelid but also results in virtually nonexistent ill effects. PMID- 17116785 TI - Long-term efficacy of biomodeled polymethyl methacrylate implants for orbitofacial defects. AB - OBJECTIVE: To report the long-term efficacy of custom polymethyl methacrylate implants using high-resolution computed tomographic modeling in the reconstruction of complex orbitofacial defects secondary to trauma. METHODS: Nine patients with complex orbitofacial bone defects after trauma were evaluated for this retrospective, nonrandomized, noncomparative study. All the patients underwent reconstruction using custom, heat-cured polymethyl methacrylate implants. Patients were followed up postoperatively and evaluated for complications. RESULTS: Nine consecutive patients (5 men and 4 women) aged 28 to 63 years who underwent surgical reconstruction using prefabricated, heat-cured polymethyl methacrylate implants were included in the study. The interval between injury and presentation ranged from 1 month to 40 years. There were no significant complications, including infection, extrusion, or displacement of the implant. In all of the patients, wound healing was uneventful, with antibiotic drugs administered perioperatively. Mean follow-up was 4.3 years from the first visit (range, 6 months to 10 years). CONCLUSIONS: Computed tomographic biomodeled, prefabricated, heat-cured polymethyl methacrylate implants are well tolerated in the long term. Their advantages include customized design, long-term biocompatibility, and excellent aesthetic results. PMID- 17116786 TI - Comparison of tensile strength of resorbable plating systems used in monocortical mandible angle osteotomy repair. AB - OBJECTIVE: To determine the usefulness of resorbable plating systems in load bearing applications of the mandible and the location of critical failure. METHODS: An osteotomy was created in 24 fresh cadaveric mandibles at the angle and fixated by the Champy technique with similar resorbable craniofacial plating systems from 4 manufacturers. Each mandible was held rigid as a material test system applied a downward force anteriorly. The critical tolerance was measured and the type of failure was noted. RESULTS: Critical failure occurred at forces from 34.6 to 137.8 N. We found a statistically significant difference between the plating groups (P<.001 for all comparisons). The point of failure was almost uniformly at the plate. CONCLUSIONS: Critical failure was overwhelmingly due to rupture of the plate rather than to stripping or shearing of the screws as had been strongly expected. We found differences in plate strengths for this particular application and did not evaluate their respective long-term resorptive properties. We do not advocate that single resorbable plate fixation be the sole means of mandible angle fracture fixation, regardless of the plating system used. PMID- 17116787 TI - Seagull wing graft: a technique for the replacement of lower lateral cartilages. AB - OBJECTIVES: To present and evaluate outcomes with the seagull wing technique, which was designed to replace the lower lateral cartilages and to reconstruct the nasal tip. METHODS: The seagull wing technique is illustrated and described in detail. Sixty patients who underwent surgery 1981 and 2002 were retrospectively evaluated. A preoperative diagnosis of tip deformities was made based on photographs, which were compared with the postoperative results. Patient satisfaction was subjectively evaluated. RESULTS: The postoperative results showed a significant statistical improvement in the following tip deformities: underprojected tip (P<.001), poorly defined tip (P<.001), alar retraction (P<.001), alar pinch (P<.001), overrotated tip (P<.001), and tip ptosis (P<.01). The average follow-up period was 14.6 months. Fifty-five patients (92%) stated that they were very satisfied with the surgical results; they required no additional revision surgery. CONCLUSIONS: The seagull wing technique is a safe and efficient reconstructive treatment for the aesthetic and functional problems that were caused by the overresection of the nasal tip cartilaginous framework. The outcomes were pleasing, and the results were stable during the follow-up period. Patient satisfaction was high. PMID- 17116788 TI - Modification of the Zitelli bilobed flap: a comparison of flap dynamics in human cadavers. AB - OBJECTIVE: To investigate the effects on overall flap dynamics of altering the length of the primary flap in the Zitelli bilobed flap procedure, because the standard Zitelli design can result in distortion of the alar margin after repair of cutaneous nasal alar defects. METHODS: Identical 1.5-cm circular cutaneous defects were made on the nasal alae and cheeks of 6 fresh human cadavers. A rectangle was inked around each cheek defect in a standardized fashion. A standard-design Zitelli bilobed flap was used to close the nasal and cheek defects on 1 side of each specimen (n = 6). The contralateral defect was closed with a modified design in which the primary flap was either 10% longer (n = 3) or 10% shorter (n = 3) than the standard primary flap as measured from the pivot point. Alar margin retraction and cheek defect distortion were measured from standardized photographs obtained before and after the repair. RESULTS: The long flap design resulted in alar retraction that was 1.33 mm less in the nasal reconstructions (P = .02) and distal defect distortion that was 2.17 mm less in the cheek reconstructions (P = .01) compared with the standard Zitelli bilobed flap. The short-flap design caused more distortion than did the standard design for both types of defect. CONCLUSION: Lengthening the primary flap in the Zitelli bilobed flap design may reduce distal wound distortion and alar retraction in the closure of certain cutaneous defects. PMID- 17116790 TI - Measurement of preoperative and postoperative nasal tip projection and rotation. AB - OBJECTIVE: To measure the effect of columellar struts and cephalic trim on tip projection and tip rotation using digitized photographs. METHODS: Using photographs of 62 patients who underwent external rhinoplasty, we retrospectively analyzed nasal tip projection (the Goode method) and rotation (nasolabial angle) before and after surgery. A cartilaginous strut was used in 36 patients, whereas 26 patients did not receive a strut. Patients were categorized into 4 subgroups, depending on the placement of a strut (placement, strut+ vs nonplacement, strut-) and the removal of the cephalic margin (removal, cephalic+ vs nonremoval, cephalic-) of the lateral crus: strut-/cephalic-, n = 17; strut+/cephalic-, n = 23; strut-/cephalic+, n = 9; strut+/cephalic+, n = 12. RESULTS: Nasal tip projection, measured with the Goode method, increased from 0.58 to 0.60 (P = .02) in the strut+ group; in the strut- group, nasal tip projection did not change significantly. Nasolabial angle increased from 93.96 degrees to 100.92 degrees in the strut+/cephalic- group and from 88.30 degrees to 95.06 degrees in the strut+/cephalic+ group. Removal of the cephalic margin alone (strut-/cephalic+) hardly affected tip rotation (P = .05). CONCLUSIONS: The external rhinoplasty approach did not lead to a decrease in nasal tip projection. A cartilaginous strut slightly increased nasal tip projection and also increased nasal tip rotation. This effect was accentuated by the removal of the cephalic margin of the lateral crus. PMID- 17116791 TI - Evaluation of hydroxyapatite cement for frontal sinus obliteration after mucocele resection. AB - OBJECTIVES: To retrospectively evaluate our experience with frontal sinus obliteration using hydroxyapatite cement (BoneSource; Stryker Biotech Europe, Montreux, Switzerland) and compare it with fat obliteration over the approximate same period. Frontal sinus obliteration with hydroxyapatite cement represents a new technique for obliteration of the frontal sinus after mucocele resection. METHODS: Exploration of the frontal sinus was performed using bicoronal, osteoplastic flaps, with mucosal removal and duct obliteration with tissue glue and muscle or fascia. Flaps were elevated over the periorbita, and Silastic sheeting was used to protect the BoneSource material from exposure as it dried. The frontal table was replaced when appropriate. RESULTS: Sixteen patients underwent frontal sinus obliteration with fat (fat obliteration group), and 38 patients underwent obliteration with BoneSource (BoneSource group). Fat obliteration failed in 2 patients, who underwent subsequent BoneSource obliteration, and none of the patients in the BoneSource group has required removal of material because of recurrent complications. Frontobasal trauma (26 patients [68%] in the BoneSource group and 9 patients [56%] in the fat obliteration group) was the most common history of mucocele formation in both groups. Major complications in the BoneSource group included 1 patient with skin fistula, which was managed conservatively, and 1 patient with recurrent ethmoiditis, which was managed surgically. Both complications were not directly attributed to the use of BoneSource. Contour deficit of the frontal bone occurred in 1 patient in the fat obliteration group and in none in the BoneSource group. Two patients in the fat obliteration group had donor site complications (hematoma and infection). Thirteen patients in the BoneSource group had at least 1 prior attempt at mucocele drainage, and no statistical relation existed between recurrent surgery and preservation of the anterior table. CONCLUSION: Hydroxyapatite is a safe, effective material to obliterate frontal sinuses infected with mucoceles, with minimal morbidity and excellent postoperative contour. PMID- 17116792 TI - Dual-porosity expanded polytetrafluoroethylene implants for lip, nasolabial groove, and melolabial groove augmentation. AB - OBJECTIVE: To evaluate the clinical outcomes with the use of a dual-porosity expanded polytetrafluoroethylene implant for midfacial rejuvenation. DESIGN: An institutional review board-approved retrospective chart review was conducted of all patients who underwent implantation with the dual-porosity expanded polytetrafluoroethylene implant between 2001 and 2005. RESULTS: A total of 170 patients, with 612 implants, were evaluated. Only 8 patients had minor complications, 3 of which necessitated implant removal. The overall results of independent observer analysis of outcomes were favorable in the majority of cases. CONCLUSION: The dual-porosity expanded polytetrafluoroethylene implant is safe and reliable to use for midfacial implantation. PMID- 17116793 TI - Long-term effects of botulinum toxin type A (Botox) on facial lines: a comparison in identical twins. AB - OBJECTIVE: To evaluate the presence of imprinted facial lines in identical twin sisters, one of whom had received botulinum toxin type A (Botox) treatment in the forehead and glabellar region regularly for 13 years and one of whom had not. Crow's feet were also compared. METHODS: One twin received Botox in the forehead and glabellar region (approximately 2 to 3 times each year over the past 13 years) and in the crow's feet (twice in past 2 years). Her twin received Botox only twice (in the forehead and glabellar region, 3 and 7 years ago). RESULTS: Imprinted forehead and glabellar lines were not evident in the regularly treated twin but were evident in the minimally treated twin. Crow's feet were less noticeable when the regularly treated twin smiled (even at 7 months after treatment) than when the minimally treated twin smiled. Untreated facial areas (eg, nasolabial folds) showed comparable aging in both twins. Neither twin experienced adverse effects. CONCLUSIONS: Long-term treatment with Botox can prevent the development of imprinted facial lines that are visible at rest. Botox treatment can also reduce crow's feet. Treatment is well tolerated, with no adverse events reported during 13 years of regular treatment in this study. PMID- 17116794 TI - An alternative method of middle vault reconstruction. AB - Surgery of the nasal valves is a challenging aspect of rhinoplasty surgery. The middle nasal vault assumes an important role in certain aspects of nasal valve collapse. Techniques that address pathologies of the middle vault include the placement of spreader grafts and the butterfly graft. We present an alternative technique of middle vault reconstruction that allows simultaneous repair of nasal valve collapse and creation of a smooth dorsal profile. The surgical technique is described in detail and representative cases are discussed. PMID- 17116795 TI - Ocular injury from local anesthetic injections: needle expulsion--the luer-lok allure. PMID- 17116797 TI - Simultaneous submission should not result in simultaneous publication. PMID- 17116798 TI - Long-term results of treatment for pancreaticobiliary maljunction without bile duct dilatation. AB - HYPOTHESIS: Resection of the gallbladder together with the dilated bile duct is the preferred treatment for pancreaticobiliary maljunction (PBM) with bile duct dilatation, whereas this treatment for PBM without bile duct dilatation is still controversial. DESIGN: Retrospective study of 196 patients from January 1979 to November 2004. SETTING: Two university hospitals. PATIENTS: One hundred ninety six patients with PBM, 152 (78%) with and 44 (22%) without bile duct dilatation, formed the basis of this study. MAIN OUTCOME MEASURES: The effects of cholecystectomy on long-term results in the patients without bile duct dilatation. RESULTS: Significant differences were observed in patients without bile duct dilatation: patients were older, carcinoma of the gallbladder was more prevalent (19 patients [43.2%] without dilatation vs 9 patients [5.9%] with dilatation), and pancreatic cancer and pancreatitis were also more frequent. Most of their gallbladder carcinomas were found at stage IV (63%). The outcome was very poor in stage IV, whereas 5 patients in stage I and II lived for more than 5 years after surgery. Of the 44 patients without bile duct dilatation, 23 with carcinoma of the gallbladder or pancreas died and the other 2 were lost to follow up. The remaining 19 patients were alive at the study's conclusion after cholecystectomy without bile duct resection. None of them had bile duct carcinoma at the time of surgery or during the mean follow-up period of 9 years after surgery. CONCLUSIONS: Prophylactic cholecystectomy without bile duct resection is the best treatment option for patients with PBM without bile duct dilatation. Possible association of gallbladder carcinoma should be kept in mind at the time of treatment of patients with PBM when the bile duct is not dilated. PMID- 17116799 TI - Pancreatic fistula after distal pancreatectomy: predictive risk factors and value of conservative treatment. AB - HYPOTHESIS: Predictive factors of pancreatic fistula (PF) and the value of conservative management of PF following distal pancreatectomy (DP) are poorly known. DESIGN: Case series. SETTING: A university hospital referral center. PATIENTS: From 1991 to 2003, 175 patients underwent DP with routine drainage of the pancreatic stump and postoperative repeated measures in drainage fluid. Pancreatic fistula was defined as an amylase level in surgical drainage fluid more than 5-fold the serum level after postoperative day 5, or amylase-rich fluid collection. Computed tomographic scan was only done for suspicion of abdominal collection. Conservative management of PF included percutaneous drainage of abdominal collection and total parenteral nutrition or maintaining oral feeding in some patients with low-volume PF. INTERVENTION: Conservative management of PF after DP. MAIN OUTCOME MEASURES: Incidence of PF according to indication, concomitant splenectomy, additional procedure, texture of parenchyma, location of transection (neck vs body), and technique of stump suture (stapler vs hand sewn), including elective ligation of the main duct, transfusions, and prophylactic use of octreotide. RESULTS: There was no mortality. Forty patients (23%) developed PF, which was symptomatic in 25 patients (63%); computed tomographic scan identified an abdominal collection in 26 (65%). Multivariate analysis identified 2 predictive factors for PF: no elective ligation of the main pancreatic duct (odds ratio, 2.2 [95% confidence interval, 1.0-4.7]) and transection at the body (odds ratio, 2.1 [95% confidence interval, 1.1-5.5]). If none or both predictive factors were present, the observed rate of PF was 16% and 63%, respectively. Pancreatic fistula was managed conservatively in 38 patients (95%), including percutaneous drainage in 16, and by reoperation in 2. CONCLUSIONS: Pancreatic fistula following DP is more frequent in cases of pancreatic division at the body level and no elective ligation of the main duct. Routine drainage of the pancreatic stump does not prevent postoperative abdominal collections. Conservative management of PF is successful in 95% of cases. PMID- 17116800 TI - Therapeutic potential of cardiotrophin 1 in fulminant hepatic failure: dual roles in antiapoptosis and cell repair. AB - HYPOTHESIS: Administration of cardiotrophin 1 (CT-1) can treat experimental fulminant hepatic failure (FHF). DESIGN: Rat model with FHF induced by D galactosamine (D-gal). SETTING: Fulminant hepatic failure is a rapidly progressive disease that lacks effective nonsurgical treatment. Cardiotrophin 1 is a member of the interleukin 6 family that can protect cells from damage in some animal disease models. ANIMALS: A rat model of FHF was induced by an intraperitoneal injection of D-gal (1.4 g/kg of body weight). Cardiotrophin 1 was administered at different time points after D-gal injection. RESULTS: Administration of CT-1 at 12 and 18 hours had a survival rate of 80% (12/15) and 70% (7/10), respectively, which was significantly higher than that of nontreatment (28% [5/18]). In addition, improvement of liver histologic findings, shortening of activated clotting time, and decrease in serum levels of total bilirubin and alanine aminotransferase were detected with CT-1 treatment. Administration of CT-1 decreased apoptotic cells and increased Ki-67 cells in the liver tissues. In vitro, CT-1 administration significantly decreased apoptotic cells and sequentially down-regulated the expression of proapoptotic molecules and up-regulated the expression of antiapoptotic molecules at different culture periods. D-galactosamine culture induced morphologic damage in a hepatocyte cell line, which was greatly improved by CT-1 administration. In addition, CT-1 treated cells demonstrated increased expression of glycoprotein 130 and up regulation of cyclin D1 and heat shock protein 90. CONCLUSION: Cardiotrophin 1 may improve the outcome of D-gal-induced FHF through its effects on antiapoptosis and cell repair. PMID- 17116801 TI - Influences on medical student career choice: gender or generation? AB - HYPOTHESIS: We hypothesized that increased enrollment of female medical students and different priorities of the current generation of students would be important influences on the declining interest in surgical careers. DESIGN: Students scored statements on surgical careers on 5-point Likert scales regarding agreement and whether these statements encouraged them to pursue a career in surgery. Data were analyzed using the Mann-Whitney U test. Qualitative comments were iteratively coded using a constant comparative method. SETTING: Nine US medical schools. PARTICIPANTS: A Web-based survey on the Association for Surgical Education server was e-mailed to medical students. A total of 1300 of the 1365 respondents stated their sex. MAIN OUTCOME MEASURES: The survey asked questions pertaining to surgical life, surgical residency, surgeons as influence, equity, family, and other influences. RESULTS: A total of 680 (52%) of the 1300 respondents were male. Men and women disagreed about whether surgeons lead well-balanced lives (68% and 77%, respectively) and saw this as a deterrent. A total of 35% of women (3% men; P<.001) were discouraged by a lack of female role models. Compared with students unlikely to study surgery, lower percentages of male (74% vs 65%) and female students (85% vs 58%) likely to study surgery agreed that career choice was influenced by their decision to have a family (P=.01 for men, P<.001 for women). Of medical students who agreed that their skill sets were compatible with surgical careers, similar percentages were likely (30% men vs 24% women) and unlikely (49% men vs 54% women) to study surgery. All differences between men and women were less apparent when students likely to study surgery were compared with students unlikely to study surgery. CONCLUSIONS: The decision to have a family was a more significant influence for women than men, but family and lifestyle priorities were also important to male students, supporting our hypothesis that generation and gender are both important influences on career choices. PMID- 17116802 TI - MKK4 status predicts survival after resection of gastric adenocarcinoma. AB - HYPOTHESIS: Lack of expression of the tumor-suppressor gene MKK4 is significantly correlated with poor survival after resection of gastric adenocarcinoma. DESIGN: Retrospective review of medical records after construction and immunolabeling of tissue microarrays for clinical correlation. SETTING: The Johns Hopkins Hospital, Baltimore, Md. PATIENTS: Patients operated on because of gastric adenocarcinoma between 1983 and 1995. Main Outcome Measure Long-term survival and MKK4 status. RESULTS: Primary tumors (N = 124) were scored as 0 (no labeling), 1+ (weak labeling), or 2+ (strong labeling) in 9 (7%), 80 (65%), and 35 (28%) patients, and 5-year survival in these patients was 0%, 21%, and 28%, respectively. Given the small size (7%) of the MKK4-negative group (as expected, given the 5%-10% incidence of genetic loss in carcinomas), a Cox proportional hazards analysis was performed, adjusting for age, sex, and tumor stage. This multivariate analysis revealed a 5-fold increased risk of death (P<.001) in patients whose primary tumors were MKK4-negative. Furthermore, the addition of MKK4 status significantly improved the Cox model, changing log likelihood from -1410 to -369, confirming that MKK4 status was truly the effector of the survival difference and not a bystander. CONCLUSIONS: The lack of expression of the tumor-suppressor gene MKK4 in resected gastric adenocarcinoma is robustly associated with poor survival. This finding may provide a useful prognostic tool in patients with gastric adenocarcinoma. PMID- 17116803 TI - Hydatid disease of the liver: a continuing surgical problem. AB - HYPOTHESIS: To study the results of different surgical strategies in the treatment of liver hydatid disease. DESIGN: A retrospective study of 287 patients. SETTING: A university hospital in Athens, Greece. PATIENTS: Two hundred eighty-seven patients with liver hydatid cysts treated surgically 1977-2004. The cysts were located in the right hepatic lobe in 192 cases, in the left lobe in 66 cases, and in both lobes in 29 cases. Eleven patients had concomitant cysts in other organs and 12 patients had multiple intra-abdominal cysts. RESULTS: All patients were treated surgically. Surgical procedures included external drainage, simple closure, marsupialization, partial cystectomy with omentoplasty, radical procedures, laparoscopic drainage, and radiofrequency ablation. The first 3 techniques carried a higher complication rate (36.5%) compared with the other techniques (17.85%; P<.05). However, omentoplasty and external drainage carried a higher recurrence rate (7.42% overall), as compared with radical procedures (3.22%). CONCLUSION: Omentoplasty and radical procedures carry a lower complication rate compared with the formerly used marsupialization and external drainage. Conservative techniques, such as omentoplasty and/or other procedures, offer an effective control of hepatic hydatidosis and are preferred over radical procedures, when possible. PMID- 17116804 TI - Implementing 1-dose antibiotic prophylaxis for prevention of surgical site infection. AB - HYPOTHESIS: Replacing a 24-hour regimen with a 1-dose antibiotic prophylaxis for elective surgery would not increase rates of surgical site infection and would decrease costs. DESIGN AND SETTING: Before-after trial in a tertiary, private general hospital in Ribeirao Preto, Sao Paulo, Brazil. PATIENTS: Surgery was performed on 6140 consecutive patients from February 2002 through October 2002 (period 1) and 6159 consecutive patients from December 2002 through August 2003 (period 2). Studied surgeries included orthopedic, gastrointestinal, urology, vascular, lung, head and neck, heart, gynecologic, oncology, colon, neurologic, and pediatric surgeries. The study excluded patients with infection at the time of surgery. INTERVENTION: Decreasing the 24-hour prophylactic antibiotic regimen to 1-dose antibiotic prophylaxis. MAIN OUTCOME MEASURES: Surgical site infections in both periods measured by in-hospital surveillance and postdischarge surveillance; compliance with 1-dose prophylaxis; and costs with cephazolin. RESULTS: We followed up 12,299 patients during their hospital stay; postdischarge surveillance increased significantly from 2717 patients (44%) to 3066 patients (50%, P<.001). One-dose prophylaxis was correctly followed in 6123 patients (99% compliance). The rate of surgical site infection did not change in either period (2% and 2.1% respectively, P = .67). The number of cephazolin vials purchased monthly decreased from 1259 to 467 with a corresponding monthly savings of $1980. CONCLUSIONS: One-dose antibiotic prophylaxis did not lead to an increase in rates of surgical site infection and brought a monthly savings of $1980 considering cephazolin alone. High compliance to 1-dose prophylaxis was achieved through an educational intervention encouraged by the hospital director and administrative measures that reduced access to extra doses. PMID- 17116805 TI - The impact of age and Medicare status on bariatric surgical outcomes. AB - HYPOTHESIS: Medicare status and increasing age are associated with poor outcomes from bariatric surgical procedures. DESIGN: Survey. SETTING: National sample of hospitalized patients in the United States. PATIENTS AND INTERVENTION: Adult patients undergoing bariatric surgery in 2001 and 2002. MAIN OUTCOME MEASURES: Mortality and adverse events. RESULTS: We assessed 25 428 bariatric procedures with logistic regression, finding that age (odds ratio, 1.04; 95% confidence interval, 1.02-1.07), male sex (odds ratio, 2.45; 95% confidence interval, 1.48 4.03), electrolyte disorders (odds ratio, 13.91; 95% confidence interval, 8.29 23.33), and congestive heart failure (odds ratio, 4.96; 95% confidence interval, 2.52-9.77) were independent risk factors for bariatric surgery mortality. Adverse outcomes increased as a function of age in a nearly linear fashion, with a steep increase after the age of 65 years. Most Medicare patients undergoing these operations were younger than 65 years and had a much greater disease burden than non-Medicare patients. CONCLUSIONS: Age, male sex, electrolyte disorders, and congestive heart failure were independent risk factors for bariatric surgical mortality. Limiting bariatric surgical procedures to those younger than 65 years is warranted because of the high morbidity and mortality associated with these operations in older patients. PMID- 17116806 TI - Meta-analysis in surgery: methods and limitations. AB - The growth of new knowledge continues to advance the surgical disciplines, and several types of literature reviews attempt to consolidate this expansion of information. Meta-analysis is one such method that integrates findings on the same subject from different studies. Within surgery, there is a wealth of literature on a given topic, which needs to be considered collectively. As such, meta-analyses have been performed to address issues like the use of bowel preparation for colorectal surgery and comparisons of outcomes for laparoscopic vs open surgical approaches. A basic understanding of the groundwork required for meta-analysis is fundamental toward interpreting and critiquing its results. This review provides an overview of the principles, application, and limitations of meta-analysis in the context of surgery. PMID- 17116807 TI - Cell response to surgery. AB - OBJECTIVES: To describe the profound alterations in host immunity that are produced by major surgery as demonstrated by experimental and clinical studies, and to evaluate the benefits of therapeutic strategies aimed at attenuating perioperative immune dysfunction. DATA SOURCES: A review of the English-language literature was conducted, incorporating searches of the MEDLINE, EMBASE, and Cochrane collaboration databases to identify laboratory and clinical studies investigating the cellular response to surgery. STUDY SELECTION: Original articles and case reports describing immune dysfunction secondary to surgical trauma were included. DATA EXTRACTION: The results were compiled to show outcomes of different studies and were compared. DATA SYNTHESIS: Current evidence indicates that the early systemic inflammatory response syndrome observed after major surgery that is characterized by proinflammatory cytokine release, microcirculatory disturbance, and cell-mediated immune dysfunction is followed by a compensatory anti-inflammatory response syndrome, which predisposes the patient to opportunistic infection, multiple organ dysfunction syndrome, and death. Because there are currently no effective treatment options for multiple organ dysfunction syndrome, measures to prevent its onset should be initiated at an early stage. Accumulating experimental evidence suggests that targeted therapeutic strategies involving immunomodulatory agents such as interferon gamma, granulocyte colony-stimulating factor, the prostaglandin E(2) antagonist, indomethacin, and pentoxifylline may be used for the treatment of systemic inflammatory response syndrome to prevent the onset of multiple organ dysfunction syndrome. CONCLUSIONS: Surgical trauma produces profound immunological dysfunction. Therapeutic strategies directed at restoring immune homeostasis should aim to redress the physiological proinflammatory-anti-inflammatory cell imbalance associated with major surgery. PMID- 17116808 TI - Image of the month. Splenic artery pseudoaneurysm. PMID- 17116809 TI - Image of the month. Marjolin ulcer. PMID- 17116810 TI - Patterns of recurrence in patients with melanoma after radical lymph node dissection. PMID- 17116811 TI - Increased isolation of methicillin-resistant Staphylococcus aureus in pediatric head and neck abscesses. AB - OBJECTIVE: To compare the proportion of community-associated, methicillin resistant Staphylococcus aureus (MRSA) infections in pediatric head and neck abscesses between 2 study periods. DESIGN: Retrospective case review. SETTING: Tertiary care pediatric otolaryngology practice. Patients Pediatric patients with head and neck abscesses presenting over 2 separate 2.5-year intervals: July 1999 through December 2001 and January 2002 through June 2004. INTERVENTIONS: Incision and drainage of abscess. MAIN OUTCOME MEASURES: Type and antimicrobial susceptibility of cultured organisms. RESULTS: We identified 21 abscesses in 19 patients from July 1999 through December 2001 and 32 abscesses in 32 patients from January 2002 through June 2004. Of the 21 abscesses in the first study period, 15 demonstrated pathogen growth compared with 29 of 32 abscesses in the second study period. In the first period, 6 (40%) of 15 abscesses yielded S aureus compared with 17 (58.6%) of 29 abscesses in the second period. The proportion of abscesses yielding MRSA increased from 0% (0/6) in the first study period to 64.7% (11/17) in the second study period (P<.01). All MRSA infections were considered to be community acquired. CONCLUSIONS: Our study demonstrates a statistically significant rise in the proportion of community-associated MRSA infections of the head and neck in the pediatric population at our institution. For communities where similar microbial recovery patterns exist, we suggest that a culture be obtained as soon as possible in a child presenting with a head and neck abscess to identify the organism. Until that time, the best empirical treatment is clindamycin, with other agents available if warranted by culture and sensitivity results. A treatment algorithm is presented. PMID- 17116812 TI - Total skeletal reconstruction of the nasal dorsum. AB - OBJECTIVE: To review the outcomes of rhinoplasty in patients who underwent full length dorsal reconstruction. DESIGN: Retrospective review. SETTING: Private practice facial plastic surgery clinic. Patients Thirty-seven patients in a consecutive series of 1273 rhinoplasty cases. INTERVENTIONS: In all patients, a single high-density porous polyethylene implant was used to span the full length of the nasal bridge as part of their rhinoplasty reconstruction. MAIN OUTCOME MEASURES: Retrospective review of medical charts and preoperative and postoperative photographs to identify any complications, including infection, extrusion, movement, or displacement, and examination of the aesthetic outcome. A telephone survey was performed to assess patient satisfaction regarding the appearance and sensation of the nose. RESULTS: In all cases, the full-length dorsal implant provided a smooth bridge contour without leaving an inverted V deformity or focal irregularities. There were no cases of infection or extrusion. In 2 cases, revision surgery was required to enhance cosmetic outcome. CONCLUSIONS: The high rate of patient acceptance, safety, and success of full length dorsal reconstruction in providing a natural frontal contour challenges the minimal indications for using this type of reconstruction as opposed to using smaller grafts. As with all alloplasts, long-term follow-up is required. PMID- 17116813 TI - Aesthetic outcome of transfacial sinus surgery: the patient's view. AB - OBJECTIVE: To investigate the patient's view of the cosmetic outcome of transfacial sinus surgery. DESIGN: Prospective survey of patients after transfacial sinus surgery in a tertiary referral academic otolaryngology department. SETTING: Academic outpatient clinic of otorhinolaryngology. Patients Seventy patients (52 men, 18 women; mean +/- SD age, 56.2 +/- 14.9 years) who had undergone transfacial sinus surgery more than 4 months prior to study entry. INTERVENTIONS: Standardized patient self-assessment for postoperative alteration of facial appearance and emotional impairment and standardized observer assessment by surgeons and laypersons by means of visual analogue scales. RESULTS: Seventy-nine percent of the patients rated their appearance unaltered or minimally altered after transfacial surgery, and 91% reported no or minimal cosmetic morbidity. Postoperative cosmetic morbidity was significantly more common in women, in patients with chronic disease, and in those operated on for trauma. The surgeons' assessment was significantly correlated with the patients' self-assessment of altered appearance, but not with the patients' emotional impairment. CONCLUSION: Consideration of these risk factors may help to further improve patient selection for, and patients' satisfaction with, transfacial sinus surgery. PMID- 17116814 TI - Up-regulation of peroxisome proliferator-activated receptor gamma in perennial allergic rhinitis. AB - OBJECTIVES: To investigate the expression of peroxisome proliferator-activated receptor gamma (PPAR-gamma) messenger RNA and protein and to localize the PPAR gamma protein in the nasal mucosa of patients with allergic rhinitis and control subjects. DESIGN: Prospective study. SETTING: Tertiary academic institution. Patients Twenty patients with perennial allergic rhinitis and 20 matched nonallergic patients. INTERVENTIONS: Inferior turbinate mucosa samples were obtained from 20 patients with perennial allergic rhinitis and 20 matched nonallegic patients. Peroxisome proliferator-activated receptor gamma messenger RNA was extracted from the inferior turbinate mucosae, and then reverse transcription-polymerase chain reaction was performed. Western blot testing was used to analyze differences in PPAR-gamma protein expression levels between patients with allergic rhinitis and normal controls, and the PPAR-gamma protein was localized immunohistochemically. RESULTS: The expression levels of PPAR-gamma messenger RNA and protein in the nasal mucosa was significantly increased in patients with perennial allergic rhinitis compared with controls. Peroxisome proliferator-activated receptor gamma protein was expressed in the epithelium, infiltrating inflammatory cells, and submucosal glands. CONCLUSIONS: Peroxisome proliferator-activated receptor gamma is expressed in the human nasal mucosa and is up-regulated in perennial allergic rhinitis. These results suggest a possible contribution for PPAR-gamma in chronic inflammation of the nasal mucosa in perennial allergic rhinitis. PMID- 17116815 TI - Visualization of flow resistance in physiological nasal respiration: analysis of velocity and vorticities using numerical simulation. AB - OBJECTIVES: To visualize the velocity gradients and the vorticities of physiological unsteady nasal flow using the computational fluid dynamics method and to compare the inspiratory phase and expiratory phase flow patterns. DESIGN: An anatomically correct 3-dimensional nasal and pharyngeal cavity was constructed from computed tomographic images of a healthy adult nose and pharynx. The unsteady state Navier-Stokes and continuity equations were solved numerically on inspiratory and expiratory nasal flow. SETTING: Numerical simulation application. PARTICIPANTS: Coronary and axial computed tomographic images from a healthy adult were used. MAIN OUTCOME MEASURES: The detailed velocity distribution and vorticity (resistance) distribution of nasal airflow were visualized using the computational fluid dynamics method (an imaging technology for regional flow factors [velocity, vector, streamline, and vortex]). RESULTS: In the inspiratory phase, a high-velocity area was prominent in the middle meatus, and the highest vorticity area had good agreement with this region. In the expiratory phase, the distributions of velocity and vorticities were flatter than those in the inspiratory phase. CONCLUSION: The computational fluid dynamics model allows the investigation of airflow elements under physiological conditions, as well as the examination of the effect of nasal structure. PMID- 17116816 TI - Estimated cost-effectiveness of active middle-ear implantation in hearing impaired patients with severe external otitis. AB - OBJECTIVE: To determine the cost-effectiveness of middle-ear implantations in hearing-impaired patients with severe external otitis in the Netherlands. DESIGN: Cost-effectiveness analysis, using single-subject repeated measures of quality of life and total cost determinations. SETTING: Hospital based. Patients Moderately to severely sensorineurally hearing-impaired patients (n = 21) with severe chronic external otitis, eligible to receive a middle-ear implant. Main Outcome Measure Cost per quality-adjusted life-year (QALY), based on scores of the Medical Outcomes Study Short-Form Health()Survey (SF-36) generic quality of life questionnaire. Only direct costs were included in cost calculation of middle-ear implantation. RESULTS: Mean health utility gain was 0.046 (0.012-0.079) (P = .01) measured at the mental component of the SF-36. With a mean profitable time of 19.4 years and an overall cost of euro 14,354, minimal cost-effectiveness of middle-ear implantation was euro 16,085/QALY. CONCLUSION: Based on the cost per QALY, middle-ear implantation proved to be a cost-effective and justified health care intervention in the Netherlands. PMID- 17116817 TI - Decreasing rates of middle ear surgery in Western Australian children. AB - OBJECTIVE: To investigate temporal, social, demographic, and health care utilization factors associated with myringotomy with ventilation tube insertion (MVTI) in Western Australian (WA) children. DESIGN: Observational retrospective population-based cohort study using hospital administrative data. SETTING: All WA hospitals. PARTICIPANTS: A total of 53 673 children younger than 15 years who underwent surgery for MVTI in the period 1981-2004. MAIN OUTCOME MEASURES: Age specific incidence rates and incidence rate ratios. RESULTS: The rate of MVTI in children younger than 15 years peaked in 1997 at 6.7 per 1000 person-years and decreased to 5.6 per 1000 person-years by 2004. Based on 2004 rates, 8.4% of WA children will undergo at least 1 MVTI procedure before reaching age 15 years. The rate of MVTI was 37% lower in Indigenous children, and the procedures were performed at an older age compared with non-Indigenous children. Higher rates of MVTI were associated with areas of higher economic resources, lower education and occupation status, and living in metropolitan areas. CONCLUSIONS: The rate of MVTI in WA is showing evidence of a decline, even among children younger than 5 years. There remains an issue regarding equity of access to care for Indigenous children. Increasing parental economic resources may be associated with higher rates of MVTI independent of educational status. PMID- 17116818 TI - Functional outcome of supracricoid partial laryngectomy with cricohyoidopexy: radiation failure vs previously untreated cases. AB - OBJECTIVE: To evaluate the postoperative course and functional outcomes achieved in patients treated with supracricoid partial laryngectomy (SPL) with cricohyoidopexy. DESIGN: Retrospective analysis. SETTING: National Cancer Institute "Regina Elena." PATIENTS: Eighty-two consecutive patients who underwent SPL with cricohyoidopexy between September 1, 1988, and June 30, 2005, were evaluated. The patient cohort was divided into 2 groups: one affected by untreated laryngeal cancer and the other with laryngeal recurrence after radiotherapy. MAIN OUTCOME MEASURES: The postoperative complications and functional outcomes of both patient groups were evaluated and statistically compared. RESULTS: No statistical differences were found between the functional results of the 2 groups of patients analyzed. CONCLUSION: Although a slightly delayed recovery of physiological functions of the larynx could be termed a disadvantage of SCL with cricohyoidopexy after radiotherapy, this operation is a reliable and useful procedure for selected patients with recurrent cancer who would otherwise have been operated on and received a total laryngectomy. PMID- 17116819 TI - Vocal outcomes after laser resection of early-stage glottic cancer with adjuvant cryotherapy. AB - OBJECTIVE: To evaluate the vocal outcomes of patients with early-stage glottic carcinoma undergoing laser resection with adjuvant cryoablative therapy. DESIGN: Retrospective review. SETTING: Tertiary care center. Patients Twenty patients with early-stage glottic carcinoma. Intervention Treatment of early-stage glottic carcinoma with endoscopic carbon dioxide laser resection in conjunction with cryoablation. MAIN OUTCOME MEASURES: Disease-free survival and subjective and objective measures of posttreatment voice quality, based on serial videolaryngostroboscopy. RESULTS: There was 1 local treatment failure, with an overall mean disease-free follow-up of 32.6 months (range, 3-93 months). Carbon dioxide laser resection and cryoablative therapy were associated with a significant improvement in subjective voice quality (P<.001). Long-term dysphonia was uniformly improved vis-a-vis the pretreatment condition, even among patients with the most advanced disease undergoing the widest resections. Posttreatment web formation was not noted among 4 patients with anterior commissure involvement. CONCLUSIONS: Endoscopic laser laryngeal surgery performed in conjunction with cryotherapy for early-stage glottic carcinoma yielded excellent primary site control, while improving subjective and objective measures of voice quality. Combined laser surgery and cryotherapy is a possible alternative to radiotherapy for selected patients with early-stage glottic carcinoma who desire curative therapy, while optimizing vocal outcomes. PMID- 17116820 TI - Chemoprevention of head and neck cancer with aspirin: a case-control study. AB - OBJECTIVE: To evaluate the chemopreventive potential of aspirin against head and neck cancer. DESIGN: Hospital-based case-control study. SETTING: National Cancer Institute-designated comprehensive cancer center. Patients Individuals who received medical services at the Roswell Park Cancer Institute, Buffalo, NY, between 1982 and 1998 and who completed a comprehensive epidemiologic questionnaire. MAIN OUTCOME MEASURE: Aspirin use among 529 patients with head and neck cancer and 529 hospital-based control subjects matched by age, sex, and smoking status. RESULTS: Aspirin use was associated with a 25% reduction in the risk of head and neck cancer (adjusted odds ratio, 0.75; 95% confidence interval, 0.58-0.96). Consistent risk reductions were also noted in association with frequent and prolonged aspirin use. Further, a consistently decreasing trend in risk was noted with increasing duration of aspirin use (P(trend) = .005). Risk reduction was observed across all 5 primary tumor sites, with cancers of the oral cavity and oropharynx exhibiting greater risk reduction. When analyzed by smoking and alcohol exposure levels, participants moderately exposed to either showed a statistically significant 33% risk reduction (adjusted odds ratio, 0.67; 95% confidence interval, 0.50-0.91), whereas participants exposed to both heavy smoking and alcohol use did not benefit from the protective effect of aspirin. The reduction in risk was relatively more significant in women. CONCLUSIONS: Aspirin use is associated with reduced risk of head and neck cancer. This effect is more pronounced in individuals with low to moderate exposure to cigarette smoke or alcohol consumption. PMID- 17116821 TI - Surgical resection of cutaneous head and neck lesions: does aspirin use increase hemorrhagic risk? AB - OBJECTIVE: To determine whether the antiplatelet agent aspirin increases hemorrhagic risk in patients undergoing surgical resection of cutaneous head and neck lesions. DESIGN: Retrospective cohort study. SETTING: Regional referral center. Patients All cases of cutaneous head and neck lesions surgically resected during a 10-year period were included. MAIN OUTCOME MEASURES: Outcome measures were incidence of significant postoperative hemorrhage, defined as postoperative hematoma or hemorrhage necessitating surgical reexploration; and total postoperative hemorrhage, defined as any hemorrhage lasting longer than 4 hours despite external pressure, requiring medical review, and resulting in prolongation of the patient's hospital stay or readmission to the hospital. RESULTS: Seven hundred eleven patients (974 cases) were eligible for inclusion, of whom 320 were receiving aspirin therapy at the time of surgery. The incidence of significant postoperative hemorrhage in the aspirin and nonaspirin groups was 5 (1.6%) and 0, respectively (P = .004), and aspirin use was the only risk factor for significant postoperative hemorrhage. The incidence of total postoperative hemorrhage in the aspirin and nonaspirin groups was 7 (2.2%) and 1 (0.1%), respectively (P = .002). At multivariate analysis, aspirin use and local flap reconstruction were independent risk factors for total postoperative hemorrhage. Cases receiving aspirin therapy who also underwent local flap reconstruction were at exponential (124-fold) increased risk of total postoperative hemorrhage compared with cases with neither risk factor. CONCLUSIONS: Aspirin intake at the time of surgery to resect cutaneous head and neck lesions confers a small but statistically increased risk of postoperative hemorrhage. This risk is particularly pronounced in patients undergoing local flap reconstruction. PMID- 17116822 TI - Proton beam radiation therapy for skull base adenoid cystic carcinoma. AB - OBJECTIVE: To determine the treatment outcome and prognostic factors in patients with adenoid cystic carcinoma of the skull base treated with proton beam radiation therapy. DESIGN: Retrospective analysis. SETTING: Massachusetts General Hospital, Massachusetts Eye and Ear Infirmary, and Harvard Medical School, Boston. PATIENTS: From 1991 to 2002, 23 patients with newly diagnosed adenoid cystic carcinoma with skull base extension were treated with combined proton and photon radiotherapy. There was tumor involvement of the sphenoid sinus in 61% of patients (14), nasopharynx in 61% (14), clivus in 48% (11), and cavernous sinus in 74% (17). The extent of surgery was biopsy alone in 48% (11), partial resection in 39% (9), and gross total resection with positive margins in 13% (3). The median total dose to the primary site was 75.9 cobalt-gray equivalent. The median follow-up of all surviving patients was 64 months. MAIN OUTCOME MEASURES: Locoregional control and disease-free survival and overall survival rates. RESULTS: Tumors recurred locally in 2 patients at 33 and 68 months, respectively. No patients developed neck recurrence. Eight patients had distant metastasis as the first site of recurrence. The local control rate at 5 years was 93%. The rate of freedom from distant metastasis at 5 years was 62%. The disease-free and overall survival rates at 5 years were 56% and 77%, respectively. In multivariate analysis, significant adverse factors predictive for overall survival were change in vision at presentation (P = .02) and involvement of sphenoid sinus and clivus (P = .01). CONCLUSIONS: High-dose conformal proton beam radiation therapy results in a very encouraging local control rate in patients with adenoid cystic carcinoma of the skull base. Changes in vision at presentation and tumor involvement of the sphenoid sinus and clivus are important prognostic factors. PMID- 17116823 TI - Radiofrequency ablation of microcystic lymphatic malformation in the oral cavity. AB - OBJECTIVE: To determine the efficacy and safety of radiofrequency (RF) ablation of vesicles and the resulting symptomatic control of microcystic lymphatic malformation (LM) in the oral cavity. DESIGN: An institutional review board approved retrospective study with follow-up telephone interview. SETTING: Tertiary pediatric medical center. Patients Eleven children (6 girls and 5 boys), aged 4 to 16 years, presenting between August 1, 2002, and December 1, 2004. Intervention Radiofrequency ablation of LM in the oral cavity. MAIN OUTCOME MEASURES: Symptoms related to LM, postoperative oral intake, and postoperative antibiotic requirements. RESULTS: Eleven patients presented with microcystic LM involving the lips, tongue, floor of the mouth, or buccal mucosa. Complaints included bleeding, infection, swelling, vesicle formation, and malocclusion. Patients underwent RF ablation (coblation) of oral cavity lesions. Seven (64%) of the 11 patients were able to tolerate oral intake in the recovery room. The need for antibiotics was reduced after RF ablation. All patients related diminished bleeding, pain, infection, or vesicle formation, with more than half reporting a significant improvement (6 patients) or complete resolution (1 patient). Five (62%) of 8 parents stated that the improvement after RF ablation was superior to that following previous procedures. CONCLUSIONS: Subtotal RF ablation of LM appears to be safe, with early postoperative oral intake and minimal postoperative pain. Further studies are needed to determine long-term control of LM. PMID- 17116824 TI - Interdigitating dendritic cell sarcoma of cervical lymph nodes. PMID- 17116825 TI - An infrequent mass of the middle ear: salivary gland choristoma. PMID- 17116826 TI - Radiology quiz case 1. Stafne bone cyst. PMID- 17116827 TI - Radiology quiz case 2. Bilateral petrous apex cephaloceles (PACs). PMID- 17116828 TI - Pathology quiz case 1. Epithelioid synovial sarcoma (synoviosarcoma). PMID- 17116829 TI - Pathology quiz case 2. Extramedullary plasmacytoma (EMP) of the nasopharynx. PMID- 17116830 TI - Successful use of rituximab for cutaneous vasculitis. PMID- 17116831 TI - Livedoid vasculopathy: further evidence for procoagulant pathogenesis. AB - OBJECTIVE: To further characterize the clinical and pathologic features, disease associations, and laboratory abnormalities of livedoid vasculopathy. DESIGN: Retrospective study of patients identified from our institutional database from January 1, 1990, to December 31, 2000. SETTING: Tertiary care institution. Patients Forty-five patients with biopsy-proved livedoid vasculopathy. MAIN OUTCOME MEASURES: Clinical presentation, histopathologic diagnosis, results of testing for coagulation abnormalities, and assessment of vascular status. RESULTS: Thirty-two patients (71.1%) were female (mean age, 45 years; age range, 10-85 years). Bilateral lower extremity disease occurred in 36 patients (80.0%), ulceration in 31 (68.9%), and atrophie blanche in 32 (71.1%). In patients tested, transcutaneous oximetry measurements were decreased in 20 (74.1%) of 27, and factor V Leiden mutation (heterozygous) was noted in 2 (22.2%) of 9, decreased activity for protein C or protein S in 2 (13.3%) of 15, prothrombin G20210A gene mutation in 1 (8.3%) of 12, and lupus anticoagulant in 5 (17.9%) of 28. Anticardiolipin antibodies were present in 8 (28.6%) of 28 patients, and elevated homocysteine levels in 3 (14.3%) of 21. Intraluminal thrombosis was observed in 44 (97.8%) of 45 skin biopsy specimens. Direct immunofluorescence disclosed multiple vascular conjugates in 31 (86.1%) of 36 biopsy specimens. CONCLUSIONS: Livedoid vasculopathy was predominantly bilateral, affected the lower extremities, and was associated with ulceration and atrophie blanche. Histologic evidence of intraluminal thrombosis was observed in almost all biopsy specimens reviewed. Laboratory testing revealed numerous heterogeneous coagulation abnormalities, providing further evidence of procoagulant mechanisms. PMID- 17116832 TI - The relationship between melanoma thickness and time to diagnosis in a large population-based study. AB - OBJECTIVE: To examine the relationship between melanoma thickness and reported time from first recognition and from first physician contact to the diagnosis of invasive melanoma. DESIGN: Telephone survey of patients recently diagnosed as having melanoma, combined with relevant pathological data (including melanoma thickness and morphologic structure) from the population-based Queensland Cancer Registry. A test-retest study (n = 176) was also conducted. SETTING: Population based study in Queensland. PARTICIPANTS: Residents of Queensland (n = 3772) who had been diagnosed as having invasive melanoma between January 1, 2000, and December 31, 2003. MAIN OUTCOME MEASURES: Prepresentation time (time between first noticing a suspicious spot and the first physician visit), postpresentation time (time between the first physician visit and diagnosis), and total time to diagnosis (time from initial detection of the melanoma to diagnosis). RESULTS: With 1 exception, we found no significant association between melanoma thickness and reported time to diagnosis for all melanomas combined, superficial spreading melanomas, or nodular melanomas. The exception was a positive association between melanoma thickness and postpresentation delay of physician-detected nodular melanomas. The reliability study gave intraclass correlation coefficients of 0.85 to 0.90 for the measures of intervals. CONCLUSIONS: This large study demonstrates no clear relationship between the melanoma thickness when diagnosed and the time from first recognition of changes or from first physician examination to diagnosis. This may be because of varying biological characteristics of melanomas, as well as methodological limitations of retrospective studies when trying to measure this complex association. PMID- 17116833 TI - Conventional and polarized dermoscopy features of dermatofibroma. AB - OBJECTIVE: To evaluate dermoscopic features and patterns of dermatofibromas using conventional and polarized light dermoscopy. DESIGN: Dermatofibromas were imaged using conventional nonpolarized contact dermoscopy (NPD), polarized contact dermoscopy (PCD), and polarized noncontact dermoscopy, followed by evaluation and comparison of dermoscopic features of the lesions. SETTING: Dermatology clinic specializing in pigmented lesions. Patients Fifty patients with dermatofibromas. RESULTS: The most common features of dermatofibromas observed with NPD and PCD were central white scarlike patches (37 [74%] and 42 [84%], respectively), brown globulelike structures (21 [42%] and 22 [44%]), vascular structures (24 [48%] and 22 [44%]), and a peripheral fine pigmented network (36 [72%] for both). A newly described feature observed with PCD was a central white patch characterized by shiny white streaks. With polarized noncontact dermoscopy, the most characteristic feature was a central pink hue or "vascular blush" (44 [88%]) and visibility of blood vessels (41 [82%]). The most common pattern identified with NPD and PCD was the combination of a peripheral pigmented network and a central white patch in 28 (56%) and 31 (62%) of lesions, respectively. With polarized noncontact dermoscopy, the most common pattern was a central pink hue with a peripheral pigmented network (23 [46%]). There was good to excellent agreement when comparing NPD with PCD images, but there was a variable level of agreement when polarized noncontact dermoscopy images were compared with NPD and PCD images. CONCLUSIONS: Conventional and polarized light dermoscopy are not equivalent but may be complementary. This study highlights some salient differences. We were able to identify new dermoscopic features and patterns not previously described with conventional dermoscopy. These new criteria can aid in the diagnosis of dermatofibroma. PMID- 17116834 TI - Imiquimod treatment of anal intraepithelial neoplasia in HIV-positive men. AB - OBJECTIVE: To evaluate the treatment of anal intraepithelial neoplasia (AIN) with the local immune response modifier imiquimod in human immunodeficiency virus (HIV)-positive men who have sex with men (MSM). DESIGN: Prospective, nonrandomized, open-label pilot study, with a mean follow-up time of 9(1/2) months. SETTING: Dermatology department of a university hospital. Patients Twenty eight consecutive HIV-positive MSM with histologically confirmed perianal (n = 23) or intra-anal (n = 5) AIN. Intervention Overnight treatment with self-applied imiquimod cream (perianal AIN) or suppositories (intra-anal AIN) 3 times a week for 16 weeks. MAIN OUTCOME MEASURES: Response to treatment was documented using clinical, cytologic, and histologic criteria. Human papillomavirus (HPV) typing and HPV DNA load determination for the high-risk HPV types 16, 18, 31, and 33 were performed. RESULTS: Seventeen (61%) of all 28 patients included in the study and 17 (77%) of the 22 patients with AIN, who applied imiquimod as instructed, showed clinical and histologic clearance at the end of therapy. Four patients had residual AIN and 1 patient did not improve. Clinical response was accompanied by a sharp decline in HPV DNA loads and by a reduction in the number of HPV types, but long-term HPV clearance was rarely achieved. In the follow-up period, AIN cleared in 3 patients with residual AIN. Fourteen (78%) of 18 imiquimod responders with at least 5 five months of follow-up had a normal cytologic and clinical picture at the end of the follow-up period. Three primary responders developed a recurrence. In 6 noncompliant patients, there was no clinical or morphological improvement and the HPV DNA loads remained high. CONCLUSIONS: Imiquimod appears to be a safe and effective treatment option for AIN in HIV positive MSM. Clinical response is accompanied by a significant decrease in high risk HPV DNA load. These results should encourage controlled randomized studies of imiquimod treatment of AIN. PMID- 17116835 TI - A comparison of oral methylprednisolone plus azathioprine or mycophenolate mofetil for the treatment of pemphigus. AB - OBJECTIVE: To investigate the safety and efficacy of oral methylprednisolone combined with azathioprine sodium or mycophenolate mofetil for the treatment of pemphigus. DESIGN: A prospective, multicenter, randomized, nonblinded clinical trial to compare 2 parallel groups of patients with pemphigus (pemphigus vulgaris and pemphigus foliaceus) treated with oral methylprednisolone plus azathioprine or oral methylprednisolone plus mycophenolate mofetil. Settings Thirteen departments of dermatology in Germany. Patients We included patients with pemphigus vulgaris (n = 33) or pemphigus foliaceus (n = 7) evidenced by clinical lesions suggestive of pemphigus, intraepidermal blistering on histological analysis of skin biopsy specimens, intercellular deposition of IgG within the epidermis, and immunoblot analysis findings for antidesmoglein 3 and/or antidesmoglein 1 autoantibodies. MAIN OUTCOME MEASURES: The cumulative total methylprednisolone doses and rate of remission. Secondary outcome measures were safety profiles and duration of remission. RESULTS: In 13 (72%) of 18 patients with pemphigus receiving oral methylprednisolone and azathioprine, complete remission was achieved after a mean +/- SD of 74 +/- 127 days compared with 20 (95%) of 21 patients receiving oral methylprednisolone and mycophenolate mofetil in whom complete remission occurred after a mean +/- SD of 91 +/- 113 days. The total median cumulative methylprednisolone dose used was 8916 mg (SD, +/-29 844 mg) in the azathioprine group compared with 9334 mg (SD, +/-13 280 mg) in the mycophenolate group. In 6 (33%) of 18 patients treated with azathioprine, grade 3 or 4 adverse effects were documented in contrast to 4 (19%) of 21 patients who received mycophenolate mofetil. Conclusion Mycophenolate mofetil and azathioprine demonstrate similar efficacy, corticosteroid-sparing effects, and safety profiles as adjuvants during treatment of pemphigus vulgaris and pemphigus foliaceus. PMID- 17116836 TI - Successful treatment of mucous membrane pemphigoid with etanercept in 3 patients. AB - BACKGROUND: Mucous membrane pemphigoid (MMP), also known as cicatricial pemphigoid, is a serious, autoimmune, blistering disorder that can result in blindness and other complications as a result of scarring of the mucous membranes. Effective treatment modalities are often toxic. Herein, we describe a novel therapeutic approach that is based on 2 reports in the literature of the successful use of etanercept to treat MMP. OBSERVATIONS: Three patients with MMP were treated with subcutaneous injections of 25 mg of etanercept twice weekly. All 3 patients had oral mucosal involvement, and 1 had severe, recalcitrant, ocular disease. Oral mucosal disease improved in all 3 patients. The patient with ocular involvement experienced stabilization of progression. CONCLUSIONS: Effective treatment modalities for MMP are often toxic. Etanercept may be an effective treatment option for MMP of the oral and ocular mucous membranes. This therapy should be considered as an alternative treatment option for patients who would require other aggressive systemic treatments, such as cyclophosphamide, corticosteroids, azathioprine sodium, and intravenous immunoglobulin. PMID- 17116837 TI - Livedoid vasculopathy associated with plasminogen activator inhibitor-1 promoter homozygosity (4G/4G) treated successfully with tissue plasminogen activator. AB - BACKGROUND: Livedoid vasculopathy (LV) is an occlusive thrombotic disease that affects primarily the small blood vessels of the lower extremities and often is associated with recurrent painful ulcerations. The pathogenesis of LV is unclear, but the disease is largely attributed to a hypercoagulable state. Factor V Leiden mutation, heterozygous protein C deficiency, homozygous hyperhomocysteinemia, and other inherited thrombophilias have been associated with LV. Plasminogen activator inhibitor-1 (PAI-1) is an important inhibitor of the fibrinolytic system. Elevated levels of PAI-1 are found in some patients with thrombotic diseases. Some of these patients are homozygous for an allele of PAI-1 containing a stretch of 4 guanines at base -675 in the promoter region. This variant is associated with elevated PAI-1 protein levels, impaired fibrinolysis, and increased risk of thrombosis. OBSERVATIONS: A 33-year-old white woman had a 3 month history of painful enlarging ulcers on both ankles. Various therapies, including administration of oral antibiotic agents and prednisone up to 100 mg/d, to treat presumed vasculitis, were unsuccessful. Skin biopsy specimens revealed numerous thick-walled small blood vessels, many of which were filled with fibrin thrombi, in association with minimal perivascular inflammatory infiltrate, extensive epidermal necrosis, and focal ulceration. A diagnosis of thrombotic vasculopathy was made. Clinical workup revealed an elevated plasma level of PAI-1 (31 microM/mL; reference range, <25 microM/mL) and PAI-1 promoter 4G/4G homozygosity detected at DNA sequencing. Treatment with heparin sodium and tissue plasminogen activator dramatically improved the lesions, resulting in complete healing of the ulcerations. Continuation of anticoagulant therapy with warfarin sodium and episodic administration of tissue plasminogen activator was required for symptomatic control. CONCLUSIONS: Patients with LV may have elevated plasma PAI-1 levels. This may be associated with the PAI-1 promoter 4G/4G genotype, which has not previously been linked with LV. Further studies in patients with LV are warranted to determine how frequently this genotype is present because it may identify responsiveness to fibrinolytic therapy. PMID- 17116838 TI - Malignant melanoma in marathon runners. AB - BACKGROUND: Marathon running has surged in popularity; it is generally believed to be healthy, but may be associated with medical risks. Over the past decade, we observed 8 ultramarathon runners with malignant melanoma. UV exposure, immunosuppression due to long-term intensive exercise, or both have been discussed as potential triggers in these patients. To further evaluate risk factors for malignant melanoma in marathon runners, we examined anamnestic, phenotypic, sun-related, and clinical variables in 210 athletes and compared them with those of an age- and sex-matched control group. OBSERVATIONS: Although control subjects exhibited higher sun sensitivity and more common melanocytic nevi, marathon runners presented with more atypical melanocytic nevi, solar lentigines, and lesions suggestive of nonmelanoma skin cancer. These findings correlated with increasing training intensity. During exercising, most runners wore shorts (96.7%) and shirts (98.6%) that would not or would only partially cover their back and extremities. Regular use of sunscreen was reported in only 56.2% of runners. CONCLUSIONS: Compared with a representative control group, marathon runners presented with an increased risk for malignant melanoma and nonmelanoma skin cancer. They should reduce UV exposure during exercising by choosing training and competition schedules with low sun exposure, wearing adequate clothing, and regularly using water-resistant sunscreens. PMID- 17116839 TI - Hair depigmentation during chemotherapy with a class III/V receptor tyrosine kinase inhibitor. AB - BACKGROUND: Hair pigmentation is regulated by several factors including the interaction of the ligand stem cell factor (SCF) with its class III receptor tyrosine kinase, c-kit. An interruption of SCF/c-kit signal transduction results in hair depigmentation. OBSERVATIONS: A 69-year-old white woman developed hair depigmentation and fine-textured hair while being treated with the phase I chemotherapeutic agent GW786034, a class III/V receptor tyrosine kinase inhibitor. Discontinuation of therapy resulted in a reversal of these hair changes. CONCLUSIONS: Treatment with oral GW786034 resulted in reversible hair depigmentation and change in hair growth rate and texture, which were most likely due to an incomplete inhibition of SCF/c-kit signaling, although the exact mechanism is unknown. It would be intriguing to investigate topical tyrosine kinase inhibitors as a treatment for unwanted body hair. PMID- 17116840 TI - Livedoid vasculopathy: what it is and how the patient should be evaluated and treated. PMID- 17116841 TI - Evaluating early detection in the diagnosis of melanoma. PMID- 17116842 TI - Dermatologists honored on stamps. PMID- 17116843 TI - An animal model of psoriasis in mice deficient in epidermal Jun proteins. PMID- 17116844 TI - Multiple eruptive angiomatous lesions in a patient with multiple myeloma. Glomeruloid hemangiomas associated with POEMS syndrome. PMID- 17116845 TI - Multiple nodules and plaques on the face and trunk. Cutaneous Rosai-Dorfman disease (RDD). PMID- 17116846 TI - Erythematous papular rash on the upper back area. Caterpillar dermatitis, or erucism. PMID- 17116847 TI - Tender papules on the hands. Idiopathic chilblains (perniosis). PMID- 17116848 TI - Transdermal therapy for erythromelalgia. PMID- 17116849 TI - Permanent filler augmentation: common complications. PMID- 17116850 TI - Pyoderma gangrenosum masquerading as dermatitis artefacta. PMID- 17116851 TI - Familial ACD with no recognizable enzyme abnormalities. PMID- 17116852 TI - Leukocytoclastic vasculitis masquerading as hand-foot syndrome in a patient treated with sorafenib. PMID- 17116853 TI - Long-term control of papular dermatitis ("dermal hypersensitivity reaction") with mycophenolate mofetil. PMID- 17116854 TI - Autoantibodies from patients with BSLE inducing recruitment of leukocytes to the dermoepidermal junction and subepidermal splits in cryosections of human skin. PMID- 17116855 TI - Cytokine-release syndrome related to rituximab limited to lesions and excision scars of lesions of primary cutaneous lymphoma. PMID- 17116856 TI - Spider spines detected by dermoscopy. PMID- 17116857 TI - Dermoscopy of venous stasis dermatitis. PMID- 17116858 TI - Acetylation of MEK2 and I kappa B kinase (IKK) activation loop residues by YopJ inhibits signaling. AB - To overcome host defenses, bacterial pathogens of the genus Yersinia inject specific effector proteins into colonized mammalian cells. One such virulence factor, YopJ, inhibits the host inflammatory response and induces apoptosis of immune cells by blocking multiple signaling pathways, including the MAPK and NF kappaB pathways. In this study, we show that YopJ exerts its deleterious effects by catalyzing the acetylation of two serine residues in the activation loop of the MAP kinase kinase, MEK2. This covalent modification prevents the phosphorylation of these serine residues that is required for activation of MEK2 and downstream signal propagation. We also show that YopJ causes acetylation of a threonine residue in the activation loop of both the alpha and beta subunits of the NF-kappaB pathway kinase, IKK. These results establish a hitherto uncharacterized mode of action for bacterial toxins and suggest the possibility that serine/threonine acetylation may occur even under nonpathogenic conditions and may be a widespread protein modification regulating protein function in eukaryotic cells. PMID- 17116859 TI - Using cryo-EM to measure the dipole potential of a lipid membrane. AB - The dipole potential of a lipid bilayer membrane accounts for its much larger permeability to anions than cations and affects the conformation and function of membrane proteins. The absolute value of the dipole potential has been very difficult to measure, although its value has been estimated to range from 200 to 1,000 mV from ion translocation rates, the surface potential of lipid monolayers, and molecular dynamics calculations. Here, a point charge probe method was used to investigate the dipole potentials of both ester and ether lipid membranes. The interactions between electrons and lipid molecules were recorded by phase contrast imaging using cryo-EM. The magnitude and the profile of the dipole potential along the bilayer normal were obtained by subtracting the contribution of the atomic potential from the cryo-EM image intensity. The peak dipole potential was estimated to be 510 and 260 mV for diphytanoylphosphatidylcholine and diphytanylphosphatidylcholine, respectively. PMID- 17116860 TI - Power spectra reveal the influence of stochasticity on nonlinear population dynamics. AB - Stochasticity alters the nonlinear dynamics of inherently cycling populations. The power spectrum can describe and explain the impacts of stochasticity. We fitted models to short observed time series of flour beetle populations in the frequency domain, then used a well fitting stochastic mechanistic model to generate detailed predictions of population spectra. Some predicted spectral peaks represent periodic phenomena induced or modified by stochasticity and were experimentally confirmed. For one experimental treatment, linearization theory explained that these peaks represent overcompensatory decay of deviations from deterministic oscillation. In another treatment, stochasticity caused frequent directional phase shifting around a cyclic attractor. This directional phase shifting was not explained by linearization theory and modified the periodicity of the system. If field systems exhibit directional phase shifting, then changing the intensity of demographic or environmental noise while holding constant the structure of the noise can change the main frequency of population fluctuations. PMID- 17116861 TI - Opposing forces of aerosol cooling and El Nino drive coral bleaching on Caribbean reefs. AB - Bleaching of corals as a result of elevated sea surface temperatures (SST) is rapidly becoming a primary source of stress for reefs globally; the scale and extent of this threat will depend on how the drivers of SST interact to influence bleaching patterns. We demonstrate how the opposing forces of the El Nino Southern Oscillation (ENSO) and levels of atmospheric aerosols drive regional scale patterns of coral bleaching across the Caribbean. When aerosol levels are low, bleaching is largely determined by El Nino strength, but high aerosol levels mitigate the effects of a severe El Nino. High aerosol levels, resulting principally from recent volcanic activity, have thus protected Caribbean reefs from more frequent widespread bleaching events but cannot be relied on to provide similar protection in the future. PMID- 17116862 TI - Stepwise binding and bending of DNA by Escherichia coli integration host factor. AB - Integration host factor (IHF) is a prokaryotic protein required for the integration of lambda phage DNA into its host genome. An x-ray crystal structure of the complex shows that IHF binds to the minor groove of DNA and bends the double helix by 160 degrees [Rice PA, Yang S, Mizuuchi K, Nash HA (1996) Cell 87:1295-1306]. We sought to dissect the complex formation process into its component binding and bending reaction steps, using stopped-flow fluorimetry to observe changes in resonance energy transfer between DNA-bound dyes, which in turn reflect distance changes upon bending. Different DNA substrates that are likely to increase or decrease the DNA bending rate were studied, including one with a nick in a critical kink position, and a substrate with longer DNA ends to increase hydrodynamic friction during bending. Kinetic experiments were carried out under pseudofirst-order conditions, in which the protein concentration is in substantial excess over DNA. At lower concentrations, the reaction rate rises linearly with protein concentration, implying rate limitation by the bimolecular reaction step. At high concentrations the rate reaches a plateau value, which strongly depends on temperature and the nature of the DNA substrate. We ascribe this reaction limit to the DNA bending rate and propose that complex formation is sequential at high concentration: IHF binds rapidly to DNA, followed by slower DNA bending. Our observations on the bending step kinetics are in agreement with results using the temperature-jump kinetic method. PMID- 17116863 TI - Speech perception problems of the hearing impaired reflect inability to use temporal fine structure. AB - People with sensorineural hearing loss have difficulty understanding speech, especially when background sounds are present. A reduction in the ability to resolve the frequency components of complex sounds is one factor contributing to this difficulty. Here, we show that a reduced ability to process the temporal fine structure of sounds plays an important role. Speech sounds were processed by filtering them into 16 adjacent frequency bands. The signal in each band was processed by using the Hilbert transform so as to preserve either the envelope (E, the relatively slow variations in amplitude over time) or the temporal fine structure (TFS, the rapid oscillations with rate close to the center frequency of the band). The band signals were then recombined and the stimuli were presented to subjects for identification. After training, normal-hearing subjects scored perfectly with unprocessed speech, and were approximately 90% correct with E and TFS speech. Both young and elderly subjects with moderate flat hearing loss performed almost as well as normal with unprocessed and E speech but performed very poorly with TFS speech, indicating a greatly reduced ability to use TFS. For the younger hearing-impaired group, TFS scores were highly correlated with the ability to take advantage of temporal dips in a background noise when identifying unprocessed speech. The results suggest that the ability to use TFS may be critical for "listening in the background dips." TFS stimuli may be useful in evaluating impaired hearing and in guiding the design of hearing aids and cochlear implants. PMID- 17116864 TI - Effect of urea on the structural dynamics of water. AB - We use polarization-resolved mid-infrared pump-probe spectroscopy to study the effect of urea on the structure and dynamics of water. Surprisingly, we find that, even at high concentrations of urea (8 M), the orientational dynamics of most water molecules are the same as in pure liquid water, showing that urea has a negligible effect on the hydrogen-bond dynamics of these molecules. However, a small fraction of the water molecules (approximately one water molecule per urea molecule) turns out to be strongly immobilized by urea, displaying orientational dynamics that are more than six times slower than in bulk water. A likely explanation is that these water molecules are tightly associated with urea, forming specific urea-water complexes. We discuss these results in light of the protein denaturing ability of aqueous urea. PMID- 17116865 TI - Photoinduced electron flow in a self-assembling supramolecular extension cable. AB - We report the design, bottom-up construction, characterization, and operation of a supramolecular system capable of mimicking the function played by a macroscopic electrical extension cable. The system is made up of a light-powered electron source, an electron drain, and a cable as the molecular components programmed to self-assemble by means of two distinct plug/socket junctions. Such connections are reversible and can be operated independently by orthogonal chemical inputs. In the source-connector-drain supermolecule, photoinduced electron transfer from source to drain occurs, and it can be switched off by dual-mode chemically controlled disassembling of the molecular components. PMID- 17116866 TI - Oncoprotein Akt/PKB induces trophic effects in murine models of Parkinson's disease. AB - Despite promising preclinical studies, neurotrophic factors have not yet achieved an established role in the treatment of human neurodegenerative diseases. One impediment has been the difficulty in providing these macromolecules in sufficient quantity and duration at affected sites. An alternative approach is to directly activate, by viral vector transduction, intracellular signaling pathways that mediate neurotrophic effects. We have evaluated this approach in dopamine neurons of the substantia nigra, neurons affected in Parkinson's disease, by adeno-associated virus 1 transduction with a gene encoding a myristoylated, constitutively active form of the oncoprotein Akt/PKB. Adeno-associated virus Myr Akt has pronounced trophic effects on dopamine neurons of adult and aged mice, including increases in neuron size, phenotypic markers, and sprouting. Transduction confers almost complete protection against apoptotic cell death in a highly destructive neurotoxin model. Activation of intracellular neurotrophic signaling pathways by vector transfer is a feasible approach to neuroprotection and restorative treatment of neurodegenerative disease. PMID- 17116867 TI - Divergent activities of an engineered antibody in murine and human systems have implications for therapeutic antibodies. AB - The MHC class I-related receptor, neonatal Fc receptor (FcRn), plays a central role in regulating the transport and in vivo persistence of immunoglobulin G (IgG). IgG-FcRn interactions can be targeted for engineering to modulate the in vivo longevity and transport of an antibody, and this has implications for the successful application of therapeutic IgGs. Although mice are widely used to preclinically test antibodies, human and mouse FcRn have significant differences in binding specificity. Here we show that an engineered human IgG1 has disparate properties in murine and human systems. The mutant shows improved transport relative to wild-type human IgG1 in assays of human FcRn function but has short in vivo persistence and competitively inhibits FcRn activity in mice. These studies indicate potential limitations of using mice as preclinical models for the analysis of engineered antibodies. Alternative assays are proposed that serve as indicators of the properties of IgGs in humans. PMID- 17116868 TI - Predicted seminal astacin-like protease is required for processing of reproductive proteins in Drosophila melanogaster. AB - During mating, males provide females with seminal fluids that include proteins affecting female physiology and, in some cases, reproductive behavior. In several species these male-derived modulators of reproduction are processed upon transfer to the female, suggesting molecular interaction between the sexes. Males could increase their reproductive success by contributing to regulation of this processing; consistent with this hypothesis, seminal fluids are rich in proteolysis regulators. However, whether these molecules carry out processing of male-derived reproductive modulators is unknown. We tested for this role using RNAi to knock down individually 11 Drosophila seminal fluid proteases and protease inhibitors. We found that CG11864, a predicted astacin-type metalloprotease in seminal fluid, is necessary to process two other seminal proteins: the ovulation hormone ovulin (Acp26Aa) and the sperm storage protein Acp36DE. This processing occurs only after all three proteins have entered the female. Moreover, CG11864 itself is processed inside males while en route to the female and before its action in processing ovulin and Acp36DE. Thus, processing of seminal proteins is stepwise in Drosophila, beginning in the male after the proteins leave their site of synthesis and continuing within another organism, the mated female, and the male-donated protease CG11864 is an agent of this latter processing. PMID- 17116869 TI - Crystal structure of HIV-1 protease in situ product complex and observation of a low-barrier hydrogen bond between catalytic aspartates. AB - HIV-1 protease is an effective target for designing drugs against AIDS, and structural information about the true transition state and the correct mechanism can provide important inputs. We present here the three-dimensional structure of a bi-product complex between HIV-1 protease and the two cleavage product peptides AETF and YVDGAA. The structure, refined against synchrotron data to 1.65 A resolution, shows the occurrence of the cleavage reaction in the crystal, with the product peptides still held in the enzyme active site. The separation between the scissile carbon and nitrogen atoms is 2.67 A, which is shorter than a normal van der Waal separation, but it is much longer than a peptide bond length. The substrate is thus in a stage just past the G'Z intermediate described in Northrop's mechanism [Northrop DB (2001) Acc Chem Res 34:790-797]. Because the products are generated in situ, the structure, by extrapolation, can give insight into the mechanism of the cleavage reaction. Both oxygens of the generated carboxyl group form hydrogen bonds with atoms at the catalytic center: one to the OD2 atom of a catalytic aspartate and the other to the scissile nitrogen atom. The latter hydrogen bond may have mediated protonation of scissile nitrogen, triggering peptide bond cleavage. The inner oxygen atoms of the catalytic aspartates in the complex are 2.30 A apart, indicating a low-barrier hydrogen bond between them at this stage of the reaction, an observation not included in Northrop's proposal. This structure forms a template for designing mechanism based inhibitors. PMID- 17116870 TI - The root endophytic fungus Piriformospora indica requires host cell death for proliferation during mutualistic symbiosis with barley. AB - Fungi of the recently defined order Sebacinales (Basidiomycota) are involved in a wide spectrum of mutualistic symbioses (including mycorrhizae) with various plants, thereby exhibiting a unique potential for biocontrol strategies. The axenically cultivable root endophyte Piriformospora indica is a model organism of this fungal order. It is able to increase biomass and grain yield of crop plants. In barley, the endophyte induces local and systemic resistance to fungal diseases and to abiotic stress. To elucidate the lifestyle of P. indica, we analyzed its symbiotic interaction and endophytic development in barley roots. We found that fungal colonization increases with root tissue maturation. The root tip meristem showed no colonization, and the elongation zone showed mainly intercellular colonization. In contrast, the differentiation zone was heavily infested by inter and intracellular hyphae and intracellular chlamydospores. The majority of hyphae were present in dead rhizodermal and cortical cells that became completely filled with chlamydospores. In some cases, hyphae penetrated cells and built a meshwork around plasmolyzed protoplasts, suggesting that the fungus either actively kills cells or senses cells undergoing endogenous programmed cell death. Seven days after inoculation, expression of barley BAX inhibitor-1 (HvBI-1), a gene capable of inhibiting plant cell death, was attenuated. Consistently, fungal proliferation was strongly inhibited in transgenic barley overexpressing GFP tagged HvBI-1, which shows that P. indica requires host cell death for proliferation in differentiated barley roots. We suggest that the endophyte interferes with the host cell death program to form a mutualistic interaction with plants. PMID- 17116871 TI - Exploring atomistic details of pH-dependent peptide folding. AB - Modeling pH-coupled conformational dynamics allows one to probe many important pH dependent biological processes, ranging from ATP synthesis, enzyme catalysis, and membrane fusion to protein folding/misfolding and amyloid formation. This work illustrates the strengths and capabilities of continuous constant pH molecular dynamics in exploring pH-dependent conformational transitions in proteins by revisiting an experimentally well studied model protein fragment, the C peptide from ribonuclease A. The simulation data reveal a bell-shaped pH profile for the total helix content, in agreement with experiment, and several pairs of electrostatic interactions that control the relative populations of unfolded and partially folded states of various helical lengths. The latter information greatly complements and extends that attainable by current experimental techniques. The present work paves the way for new and exciting applications, such as the study of pH-dependent molecular mechanism in the formation of amyloid comprising peptides from Alzheimer's and Parkinson's diseases. PMID- 17116872 TI - Phosphorylation and cleavage of presenilin-associated rhomboid-like protein (PARL) promotes changes in mitochondrial morphology. AB - Remodeling of mitochondria is a dynamic process coordinated by fusion and fission of the inner and outer membranes of the organelle, mediated by a set of conserved proteins. In metazoans, the molecular mechanism behind mitochondrial morphology has been recruited to govern novel functions, such as development, calcium signaling, and apoptosis, which suggests that novel mechanisms should exist to regulate the conserved membrane fusion/fission machinery. Here we show that phosphorylation and cleavage of the vertebrate-specific Pbeta domain of the mammalian presenilin-associated rhomboid-like (PARL) protease can influence mitochondrial morphology. Phosphorylation of three residues embedded in this domain, Ser-65, Thr-69, and Ser-70, impair a cleavage at position Ser(77)-Ala(78) that is required to initiate PARL-induced mitochondrial fragmentation. Our findings reveal that PARL phosphorylation and cleavage impact mitochondrial dynamics, providing a blueprint to study the molecular evolution of mitochondrial morphology. PMID- 17116873 TI - Two isoforms of tissue transglutaminase mediate opposing cellular fates. AB - Opposing cellular responses are typically regulated by distinct sets of genes. However, tissue transglutaminase (TGase) provides an interesting example of a single gene product that has been implicated both in affording protection against cellular insults as well as in promoting cell death. Here, we shed some light on how these conflicting activities might be manifested by demonstrating that alternative transcripts of TGase differentially affect cell viability. We show that although the full-length TGase protein affords strong protection against cell death signals, a shorter version of TGase that is truncated at the 3' end, and thus called TGase-short (TGase-S), is cytotoxic. The apoptotic activity of TGase-S is not dependent on its transamidation activity because the mutation of a cysteine residue that is essential for catalyzing this reaction does not compromise the ability of TGase-S to induce cell death. Intriguingly, TGase-S undergoes inappropriate oligomer formation in cells before cell death, suggesting a novel mechanism for the apoptotic effects of this protein. PMID- 17116874 TI - Blocking the apolipoprotein E/amyloid-beta interaction as a potential therapeutic approach for Alzheimer's disease. AB - The amyloid-beta (Abeta) cascade hypothesis of Alzheimer's disease (AD) maintains that accumulation of Abeta peptide constitutes a critical event in the early disease pathogenesis. The direct binding between Abeta and apolipoprotein E (apoE) is an important factor implicated in both Abeta clearance and its deposition in the brain's parenchyma and the walls of meningoencephalic vessels as cerebral amyloid angiopathy. With the aim of testing the effect of blocking the apoE/Abeta interaction in vivo as a potential novel therapeutic target for AD pharmacotherapy, we have developed Abeta12-28P, which is a blood-brain-barrier permeable nontoxic, and nonfibrillogenic synthetic peptide homologous to the apoE binding site on the full-length Abeta. Abeta12-28P binds with high affinity to apoE, preventing its binding to Abeta, but has no direct effect on Abeta aggregation. Abeta12-28P shows a strong pharmacological effect in vivo. Its systemic administration resulted in a significant reduction of Abeta plaques and cerebral amyloid angiopathy burden and a reduction of the total brain level of Abeta in two AD transgenic mice models. The treatment did not affect the levels of soluble Abeta fraction or Abeta oligomers, indicating that inhibition of the apoE/Abeta interaction in vivo has a net effect of increasing Abeta clearance over deposition and at the same time does not create conditions favoring formation of toxic oligomers. Furthermore, behavioral studies demonstrated that treatment with Abeta12-28P prevents a memory deficit in transgenic animals. These findings provide evidence of another therapeutic approach for AD. PMID- 17116875 TI - A critical role for TNF receptor-associated factor 1 and Bim down-regulation in CD8 memory T cell survival. AB - The mechanisms that allow the maintenance of immunological memory remain incompletely defined. Here we report that tumor necrosis factor receptor (TNFR) associated factor (TRAF) 1, a protein recruited in response to several costimulatory TNFR family members, is required for maximal CD8 T cell responses to influenza virus in mice. Decreased recovery of CD8 T cells in vivo occurred under conditions where cell division was unimpaired. In vitro, TRAF1-deficient, antigen-activated T cells accumulated higher levels of the proapoptotic BH3-only family member Bim, particularly the most toxic isoform, Bim(S). In the presence of excess IL-15, memory phenotype T cells with similar surface phenotype and comparable levels of Bcl-2 family members could be generated from WT or TRAF1 deficient T cell receptor transgenic OT-I T cells. However, when the memory CD8 T cells were allowed to compete for survival signals in the absence of antigen in vivo, the TRAF1-deficient T cells showed decreased recovery compared with TRAF1 sufficient T cells. This defect in T cell recovery in vivo was alleviated by introduction of siRNA to down-modulate Bim in TRAF1-deficient memory T cells. These studies identify the TRAF1 signaling axis and Bim down-regulation as critical for CD8 memory T cell survival in vivo. PMID- 17116876 TI - Ligation of the cell surface receptor, CD46, alters T cell polarity and response to antigen presentation. AB - Lymphocyte function in vivo is dictated by multiple external cues, but the integration of different signals is not well understood. Here, we show that competition for the axis of polarization dictates functional outcomes. We investigated the effect of ligation of the immunoregulatory cell surface receptor, CD46, on lymphocyte polarity during antigen presentation and cytotoxic effector function. Ligation of CD46 on human T cells prevented recruitment of the microtubule organizing center, CD3, and perforin to the interface with the antigen-presenting cell and caused a reduction in IFN-gamma production. In human NK cells, similar changes in polarity induced by CD46 ligation inhibited the recruitment of the microtubule organizing center and perforin to the interface with target cells and correlated with reduced killing. These data indicate that external signals can alter lymphocyte polarization toward antigen-presenting cells or target cells, inhibiting lymphocyte function. PMID- 17116877 TI - Gfi-1 plays an important role in IL-2-mediated Th2 cell expansion. AB - Enforced expression of growth factor independent 1 (Gfi-1), a transcription repressor induced by T cell activation and IL-4/Stat6 signaling, strikingly enhances Th2 cell expansion. Using conditionally Gfi1-deficient mice prepared for this study, we show that in vitro or in vivo deletion of this factor dramatically reduces Th2, but not Th1, cell expansion in response to IL-2. Both increased cell apoptosis and reduced cell proliferation resulted from Gfi1 deletion. IL-2-Stat5 signaling was partially reduced in Gfi1-deficient Th2 cells, but overexpression of Stat5 failed to restore normal Th2 expansion in these cells, suggesting that Gfi-1 also functioned downstream of, or in parallel with, Stat5 signaling. Reduced Th2 cell expansion in the absence of Gfi-1 was confirmed by the diminished frequency of IL-4-producing cells when these mice were infected with Schistosoma mansoni. PMID- 17116878 TI - High-resolution in situ x-ray study of the hydrophobic gap at the water-octadecyl trichlorosilane interface. AB - The knowledge of the microscopic structure of water at interfaces is essential for the understanding of interfacial phenomena in numerous natural and technological environments. To study deeply buried liquid water-solid interfaces, high-energy x-ray reflectivity measurements have been performed. Silicon wafers, functionalized by a self-assembled monolayer of octadecyl-trichlorosilane, provide strongly hydrophobic substrates. We show interfacial density profiles with angstrom resolution near the solid-liquid interface of water in contact with an octadecyl-trichlorosilane layer. The experimental data provide clear evidence for the existence of a hydrophobic gap on the molecular scale with an integrated density deficit rhod = 1.1 A g cm(-3) at the solid-water interface. In addition, measurements on the influence of gases (Ar, Xe, Kr, N(2), O(2), CO, and CO(2)) and HCl, dissolved in the water, have been performed. No effect on the hydrophobic water gap was found. PMID- 17116879 TI - Polymeric triple-shape materials. AB - Shape-memory polymers represent a promising class of materials that can move from one shape to another in response to a stimulus such as heat. Thus far, these systems are dual-shape materials. Here, we report a triple-shape polymer able to change from a first shape (A) to a second shape (B) and from there to a third shape (C). Shapes B and C are recalled by subsequent temperature increases. Whereas shapes A and B are fixed by physical cross-links, shape C is defined by covalent cross-links established during network formation. The triple-shape effect is a general concept that requires the application of a two-step programming process to suitable polymers and can be realized for various polymer networks whose molecular structure allows formation of at least two separated domains providing pronounced physical cross-links. These domains can act as the switches, which are used in the two-step programming process for temporarily fixing shapes A and B. It is demonstrated that different combinations of shapes A and B for a polymer network in a given shape C can be obtained by adjusting specific parameters of the programming process. Dual-shape materials have already found various applications. However, as later discussed and illustrated by two examples, the ability to induce two shape changes that are not limited to be unidirectional rather than one could potentially offer unique opportunities, such as in medical devices or fasteners. PMID- 17116880 TI - A second isoform of the ferredoxin:NADP oxidoreductase generated by an in-frame initiation of translation. AB - Ferredoxin:NADP oxidoreductases (FNRs) constitute a family of flavoenzymes that catalyze the exchange of reducing equivalents between one-electron carriers and the two-electron-carrying NADP(H). The main role of FNRs in cyanobacteria and leaf plastids is to provide the NADPH for photoautotrophic metabolism. In root plastids, a distinct FNR isoform is found that has been postulated to function in the opposite direction, providing electrons for nitrogen assimilation at the expense of NADPH generated by heterotrophic metabolism. A multiple gene family encodes FNR isoenzymes in plants, whereas there is only one FNR gene (petH) in cyanobacteria. Nevertheless, we detected two FNR isoforms in the cyanobacterium Synechocystis sp. strain PCC6803. One of them (FNR(S) approximately 34 kDa) is similar in size to the plastid FNR and specifically accumulates under heterotrophic conditions, whereas the other one (FNR(L) approximately 46 kDa) contains an extra N-terminal domain that allows its association with the phycobilisome. Site-directed mutants allowed us to conclude that the smaller isoform, FNR(S), is produced from an internal ribosome entry site within the petH ORF. Thus we have uncovered a mechanism by which two isoforms are produced from a single gene, which is, to our knowledge, novel in photosynthetic bacteria. Our results strongly suggest that FNR(L) is an NADP(+) reductase, whereas FNR(S) is an NADPH oxidase. PMID- 17116881 TI - Apoptosis-inducing factor mediates poly(ADP-ribose) (PAR) polymer-induced cell death. AB - Apoptosis-inducing factor (AIF), a mitochondrial oxidoreductase, is released into the cytoplasm to induce cell death in response to poly(ADP-ribose) (PAR) polymerase-1 (PARP-1) activation. How PARP-1 activation leads to AIF release is not known. Here we identify PAR polymer as a cell death signal that induces release of AIF. PAR polymer induces mitochondrial AIF release and translocation to the nucleus. PAR glycohydrolase, which degrades PAR polymer, prevents PARP-1 dependent AIF release. Cells with reduced levels of AIF are resistant to PARP-1 dependent cell death and PAR polymer cytotoxicity. These results reveal PAR polymer as an AIF-releasing factor that plays important roles in PARP-1-dependent cell death. PMID- 17116882 TI - Poly(ADP-ribose) (PAR) polymer is a death signal. AB - Excessive activation of the nuclear enzyme, poly(ADP-ribose) polymerase-1 (PARP 1) plays a prominent role in various of models of cellular injury. Here, we identify poly(ADP-ribose) (PAR) polymer, a product of PARP-1 activity, as a previously uncharacterized cell death signal. PAR polymer is directly toxic to neurons, and degradation of PAR polymer by poly(ADP-ribose) glycohydrolase (PARG) or phosphodiesterase 1 prevents PAR polymer-induced cell death. PARP-1-dependent, NMDA excitotoxicity of cortical neurons is reduced by neutralizing antibodies to PAR and by overexpression of PARG. Neuronal cultures with reduced levels of PARG are more sensitive to NMDA excitotoxicity than WT cultures. Transgenic mice overexpressing PARG have significantly reduced infarct volumes after focal ischemia. Conversely, mice with reduced levels of PARG have significantly increased infarct volumes after focal ischemia compared with WT littermate controls. These results reveal PAR polymer as a signaling molecule that induces cell death and suggests that interference with PAR polymer signaling may offer innovative therapeutic approaches for the treatment of cellular injury. PMID- 17116883 TI - A physical linkage between cystic fibrosis airway surface dehydration and Pseudomonas aeruginosa biofilms. AB - A vexing problem in cystic fibrosis (CF) pathogenesis has been to explain the high prevalence of Pseudomonas aeruginosa biofilms in CF airways. We speculated that airway surface liquid (ASL) hyperabsorption generates a concentrated airway mucus that interacts with P. aeruginosa to promote biofilms. To model CF vs. normal airway infections, normal (2.5% solids) and CF-like concentrated (8% solids) mucus were prepared, placed in flat chambers, and infected with an approximately 5 x 10(3) strain PAO1 P. aeruginosa. Although bacteria grew to 10(10) cfu/ml in both mucus concentrations, macrocolony formation was detected only in the CF-like (8% solids) mucus. Biophysical and functional measurements revealed that concentrated mucus exhibited properties that restrict bacterial motility and small molecule diffusion, resulting in high local bacterial densities with high autoinducer concentrations. These properties also rendered secondary forms of antimicrobial defense, e.g., lactoferrin, ineffective in preventing biofilm formation in a CF-like mucus environment. These data link airway surface liquid hyperabsorption to the high incidence of P. aeruginosa biofilms in CF via changes in the hydration-dependent physical-chemical properties of mucus and suggest that the thickened mucus gel model will be useful to develop therapies of P. aeruginosa biofilms in CF airways. PMID- 17116884 TI - Direct peptide-regulatable interactions between MHC class I molecules and tapasin. AB - Tapasin (Tpn) has been implicated in multiple steps of the MHC class I assembly pathway, but the mechanisms of function remain incompletely understood. Using purified proteins, we could demonstrate direct binding of Tpn to peptide deficient forms of MHC class I molecules at physiological temperatures. Tpn also bound to M10.5, a pheromone receptor-associated MHC molecule that has an open and empty groove and that shares significant sequence identity with class I sequences. Two types of MHC class I-Tpn complexes were detectable in vitro depending on the input proteins; those depleted in beta(2)m, and those containing beta(2)m. Both were competent for subsequent assembly with peptides, but the latter complexes assembled more rapidly. Thus, the assembly rate of Tpn associated class I was determined by the conditions under which Tpn-MHC class I complexes were induced. Peptide loading of class I inhibited Tpn-class I-binding interactions, and peptide-depletion enhanced binding. In combination with beta(2)m, certain peptides induced efficient dissociation of preformed Tpn-class I complexes. Together, these studies demonstrate direct Tpn-MHC class I interactions and preferential binding of empty MHC class I by Tpn, and that the Tpn-class I interaction is regulated by both beta(2)m and peptide. In cells, Tpn is likely to be a direct mediator of peptide-regulated binding and release of MHC class I from the TAP complex. PMID- 17116885 TI - Tipin and Timeless form a mutually protective complex required for genotoxic stress resistance and checkpoint function. AB - Tipin is a mammalian protein that interacts with Timeless, which plays a role in DNA damage checkpoint responses. Here, we show that Tipin is a nuclear protein that associates with the replicative helicase and protects cells against genotoxic agents. Tipin is required for efficient cell cycle arrest in response to DNA damage, and depletion of Tipin renders cells sensitive to ionizing radiation as well as replication stress. Loss of Tipin results in spontaneous gamma-H2AX foci, a marker for DNA double-strand breaks. We find that Tipin and Timeless form a complex that maintains the level of both proteins in cells and that the loss of either one will lead to the loss of the interacting partner. This observation explains the similar checkpoint phenotypes observed in both Tipin- and Timeless-depleted cells. PMID- 17116886 TI - Influence of dominant HIV-1 epitopes on HLA-A3/peptide complex formation. AB - The binding of peptides to MHC class I molecules induces MHC/peptide complexes that have specific conformational features. Little is known about the molecular and structural bases required for an optimal MHC/peptide association able to induce a dominant T cell response. We sought to characterize the interaction between purified HLA-A3 molecules and four well known CD8 epitopes from HIV-1 proteins. To define the characteristics of HLA-peptide complex formation and to identify potential structural changes, we used biochemical assays that detect well formed complexes. We tested the amplitude, stability, and kinetic parameters of the interaction between HLA-A3, peptides, and anti-HLA mAbs. Our results show that the four epitopes Nef73-82, Pol325-333, Env37-46, and Gag20-28 bind strongly to HLA-A3 molecules and form very stable complexes that are detected with differential patterns of mAb reactivity. The most striking result is the nonrecognition of the HLA-A3/Gag20-28 complex by the A11.1M mAb specific to HLA A3/-A11 alleles. To explain this observation, from the data published on HLA-A11 crystallographic structure, we propose molecular models of the HLA-A3 molecule complexed with Nef73-82, Pol325-333, and Gag20-28 epitopes. In the HLA-A3/Gag20 28 complex, we suggest that Arg at position P1 of the peptide may push the alpha2 helix residue Trp-167 of HLA-A3 and affect mAb recognition. Such observations may have great implications for T cell antigen receptor recognition and the immunogenicity of HLA/peptide complexes. PMID- 17116887 TI - Costs and benefits of processivity in enzymatic degradation of recalcitrant polysaccharides. AB - Many enzymes that hydrolyze insoluble crystalline polysaccharides such as cellulose and chitin guide detached single-polymer chains through long and deep active-site clefts, leading to processive (stepwise) degradation of the polysaccharide. We have studied the links between enzyme efficiency and processivity by analyzing the effects of mutating aromatic residues in the substrate-binding groove of a processive chitobiohydrolase, chitinase B from Serratia marcescens. Mutation of two tryptophan residues (Trp-97 and Trp-220) close to the catalytic center (subsites +1 and +2) led to reduced processivity and a reduced ability to degrade crystalline chitin, suggesting that these two properties are linked. Most remarkably, the loss of processivity in the W97A mutant was accompanied by a 29-fold increase in the degradation rate for single polymer chains as present in the soluble chitin-derivative chitosan. The properties of the W220A mutant showed a similar trend, although mutational effects were less dramatic. Processivity is thought to contribute to the degradation of crystalline polysaccharides because detached single-polymer chains are kept from reassociating with the solid material. The present results show that this processivity comes at a large cost in terms of enzyme speed. Thus, in some cases, it might be better to focus strategies for enzymatic depolymerization of polysaccharide biomass on improving substrate accessibility for nonprocessive enzymes rather than on improving the properties of processive enzymes. PMID- 17116888 TI - Quantitative analysis of Argonaute protein reveals microRNA-dependent localization to stress granules. AB - Argonaute proteins associate with microRNAs (miRNAs) that bind mRNAs through partial base-pairings to primarily repress translation in animals. A fraction of Argonaute proteins and miRNAs biochemically cosediment with polyribosomes, yet another fraction paradoxically accumulates in ribosome-free processing bodies (PBs) in the cytoplasm. In this report, we give a quantitative account of the Argonaute protein localization and dynamics in living cells in different cellular states. We find that the majority of Argonaute is distributed diffusely in the cytoplasm, and, when cells are subjected to stress, Argonaute proteins accumulate to newly assembled structures known as stress granules (SGs) in addition to PBs. Argonaute proteins displayed distinct kinetics at different structures: exchange faster at SGs and much slower at PBs. Further, miRNAs are required for the Argonaute protein localization to SGs but not PBs. These quantitative kinetic data provide insights into miRNA-mediated repression. PMID- 17116889 TI - Charge transfer through DNA nanoscaled assembly programmable with DNA building blocks. AB - DNA nanostructures based on programmable DNA molecular recognition have been developed, but the nanoelectronics of using DNA is still challenging. A more rapid charge-transfer (CT) process through the DNA nanoassembly is required for further development of programmable DNA nanoelectronics. In this article, we present direct absorption measurements of the long-range CT over a 140-A DNA assembly based on a GC repetitive sequence constructed by simply mixing DNA building blocks. We show that a CT through DNA nanoscale assembly is possible and programmable with the designed DNA sequence. PMID- 17116890 TI - Random mutations, selected mutations: A PIN opens the door to new genetic landscapes. PMID- 17116891 TI - Charting uncertainty about ant origins. PMID- 17116892 TI - Multiple independent origins of sex chromosomes in amniotes. PMID- 17116894 TI - A year in the life of Canadian Forces Health Services. PMID- 17116893 TI - FAPP2, cilium formation, and compartmentalization of the apical membrane in polarized Madin-Darby canine kidney (MDCK) cells. AB - We have analyzed the role of the phosphatidylinositol-4-phosphate adaptor protein 2 (FAPP2), a component of the apical transport machinery, in cilium formation in polarized Madin-Darby canine kidney (MDCK) cells. We show that ciliogenesis is defective in FAPP2 knockdown cells. Furthermore, by using fluorescence recovery after photobleaching studies of domain connectivity and the generalized polarization spectra of Laurdan, we demonstrate that FAPP2 depletion impairs the formation of condensed apical membrane domains. Laurdan staining also revealed that the ciliary membrane has a highly condensed bilayer domain at its base that could function as a fence to separate the ciliary membrane from the surrounding apical membrane. These results indicate that the compartmentalization of the apical membrane in MDCK cells into the ciliary membrane and the surrounding membrane depends on the balance of raft and nonraft domains. PMID- 17116895 TI - Afghanistan war poses unique challenges for military MDs. PMID- 17116896 TI - HIV prophylaxis expensive and sometimes difficult to obtain. PMID- 17116897 TI - Complex and unique HIV/AIDS epidemic among Aboriginal Canadians. PMID- 17116898 TI - Fight to free the world of WMD heats up. PMID- 17116899 TI - Trends in physician supply. PMID- 17116900 TI - Preparing Canadian military surgeons for Afghanistan. PMID- 17116901 TI - A child with an underrecognized form of developmental delay: a congenital disorder of glycosylation. PMID- 17116902 TI - Laboratory diagnosis of human infection with avian influenza. PMID- 17116903 TI - Silent menace: septic abdominal thrombophlebitis. PMID- 17116904 TI - Development of a triage protocol for critical care during an influenza pandemic. AB - BACKGROUND: The recent outbreaks of avian influenza (H5N1) have placed a renewed emphasis on preparing for an influenza pandemic in humans. Of particular concern in this planning is the allocation of resources, such as ventilators and antiviral medications, which will likely become scarce during a pandemic. METHODS: We applied a collaborative process using best evidence, expert panels, stakeholder consultations and ethical principles to develop a triage protocol for prioritizing access to critical care resources, including mechanical ventilation, during a pandemic. RESULTS: The triage protocol uses the Sequential Organ Failure Assessment score and has 4 main components: inclusion criteria, exclusion criteria, minimum qualifications for survival and a prioritization tool. INTERPRETATION: This protocol is intended to provide guidance for making triage decisions during the initial days to weeks of an influenza pandemic if the critical care system becomes overwhelmed. Although we designed this protocol for use during an influenza pandemic, the triage protocol would apply to patients both with and without influenza, since all patients must share a single pool of critical care resources. PMID- 17116905 TI - Changes in illicit opioid use across Canada. PMID- 17116906 TI - A bibliometric analysis by geographic area of published research in several biomedical fields, 1995-2003. AB - We summarized the findings of several studies of ours to compare the quantity and quality of published research from around the world for the years 1995 to 2003. We evaluated the number of articles published and their mean journal impact factor. We also studied the research productivity of various areas adjusted for gross domestic product (GDP) and population. We found that Western Europe leads the world in published research on infectious diseases-microbiology (82,342 articles [38.8%]) and in cardiopulmonary medicine (67,783 articles [39.5%]), whereas the United States ranks first in the fields of preventive medicine, public health and epidemiology both in quantity (23,918 articles [49.1%]) and quality of published papers. However, after adjustments for GDP, Canada ranked first, with the United States and Oceania following closely behind. All of the developing regions had only small research contributions in all of the biomedical fields examined. PMID- 17116907 TI - Pandemic triage: the ethical challenge. PMID- 17116908 TI - The need to promote public health in the field of illicit drug use. PMID- 17116909 TI - Summary of findings from the evaluation of a pilot medically supervised safer injecting facility. AB - In many cities, infectious disease and overdose epidemics are occurring among illicit injection drug users (IDUs). To reduce these concerns, Vancouver opened a supervised safer injecting facility in September 2003. Within the facility, people inject pre-obtained illicit drugs under the supervision of medical staff. The program was granted a legal exemption by the Canadian government on the condition that a 3-year scientific evaluation of its impacts be conducted. In this review, we summarize the findings from evaluations in those 3 years, including characteristics of IDUs at the facility, public injection drug use and publicly discarded syringes, HIV risk behaviour, use of addiction treatment services and other community resources, and drug-related crime rates. Vancouver's safer injecting facility has been associated with an array of community and public health benefits without evidence of adverse impacts. These findings should be useful to other cities considering supervised injecting facilities and to governments considering regulating their use. PMID- 17116912 TI - Diet and weight loss. PMID- 17116913 TI - Summaries for patients. Can a coordinated system of care improve the quality of care for people with dementia? PMID- 17116914 TI - Summaries for patients. Can providing support to dementia caregivers improve their quality of life? PMID- 17116915 TI - Summaries for patients. Relationship of insurance type with the care of acute coronary syndromes. PMID- 17116916 TI - The effect of a disease management intervention on quality and outcomes of dementia care: a randomized, controlled trial. AB - BACKGROUND: Adherence to dementia guidelines is poor despite evidence that some guideline recommendations can improve symptoms and delay institutionalization of patients. OBJECTIVE: To test the effectiveness of a dementia guideline-based disease management program on quality of care and outcomes for patients with dementia. DESIGN: Clinic-level, cluster randomized, controlled trial. SETTING: 3 health care organizations collaborating with 3 community agencies in southern California. PARTICIPANTS: 18 primary care clinics and 408 patients with dementia age 65 years or older paired with 408 informal caregivers. INTERVENTION: Disease management program led by care managers and provided to 238 patient-caregiver pairs at 9 intervention clinics for more than 12 months. MEASUREMENTS: Adherence to 23 guideline recommendations (primary outcome) and receipt of community resources and patient and caregiver health and quality-of-care measures (secondary outcomes). RESULTS: The mean percentage of per-patient guideline recommendations to which care was adherent was significantly higher in the intervention group than in the usual care group (63.9% vs. 32.9%, respectively; adjusted difference, 30.1% [95% CI, 25.2% to 34.9%]; P < 0.001). Participants who received the intervention had higher care quality on 21 of 23 guidelines (P < or = 0.013 for all), and higher proportions received community agency assistance (P < or = 0.03) than those who received usual care. Patient health-related quality of life, overall quality of patient care, caregiving quality, social support, and level of unmet caregiving assistance needs were better for participants in the intervention group than for those in the usual care group (P < 0.05 for all). Caregiver health-related quality of life did not differ between the 2 groups. LIMITATIONS: Participants were well-educated, were predominantly white, had a usual source of care, and were not institutionalized. Generalizability to other patients and geographic regions is unknown. Also, costs of a care management program under fee-for-service reimbursement may impede adoption. CONCLUSIONS: A dementia guideline-based disease management program led to substantial improvements in quality of care for patients with dementia. Current Controlled Trials identifier: ISRCTN72577751. PMID- 17116917 TI - Enhancing the quality of life of dementia caregivers from different ethnic or racial groups: a randomized, controlled trial. AB - BACKGROUND: Caring for a family member with dementia is extremely stressful, contributes to psychiatric and physical illness among caregivers, and increases the risk for caregiver death. Finding better ways to support family caregivers is a major public health challenge. OBJECTIVE: To test the effects of a structured multicomponent intervention on quality of life and clinical depression in caregivers and on rates of institutional placement of care recipients in 3 diverse racial or ethnic groups. DESIGN: Randomized, controlled trial. SETTING: In-home caregivers in 5 U.S. cities. PARTICIPANTS: 212 Hispanic or Latino, 219 white or Caucasian, and 211 black or African-American caregivers and their care recipients with Alzheimer disease or related disorders. INTERVENTION: Caregivers within each racial or ethnic group were randomly assigned to an intervention or to a control group. The intervention addressed caregiver depression, burden, self care, and social support and care recipient problem behaviors through 12 in-home and telephone sessions over 6 months. Caregivers in the control group received 2 brief "check-in" telephone calls during the 6-month intervention. MEASUREMENTS: The primary outcome was a quality-of-life indicator comprising measures of 6 month caregiver depression, burden, self-care, and social support and care recipient problem behaviors. Secondary outcomes were caregiver clinical depression and institutional placement of the care recipient at 6 months. RESULTS: Hispanic or Latino and white or Caucasian caregivers in the intervention group experienced significantly greater improvement in quality of life than those in the control group (P < 0.001 and P = 0.037, respectively). Black or African American spouse caregivers also improved significantly more (P = 0.003). Prevalence of clinical depression was lower among caregivers in the intervention group (12.6% vs. 22.7%; P = 0.001). There were no statistically significant differences in institutionalization at 6 months. LIMITATIONS: The study used only a single 6-month follow-up assessment, combined heterogeneous cultures and ethnicities into a single group, and excluded some ethnic groups. CONCLUSIONS: A structured multicomponent intervention adapted to individual risk profiles can increase the quality of life of ethnically diverse dementia caregivers. ClinicalTrials.gov identifier: NCT00177489. PMID- 17116918 TI - Insurance coverage and care of patients with non-ST-segment elevation acute coronary syndromes. AB - BACKGROUND: The impact of insurance coverage on the care of patients with non-ST segment elevation acute coronary syndromes (NSTE ACS) is unclear. OBJECTIVE: To compare NSTE ACS care patterns by insurance type. DESIGN: Comparison of Medicaid patients younger than 65 years of age and Medicare patients 65 years of age or older with patients of similar age who have health maintenance organization (HMO) or private insurance coverage. SETTING: 521 U.S. hospitals participating in the CRUSADE (Can Rapid Risk Stratification of Unstable Angina Patients Suppress ADverse Outcomes with Early Implementation of the ACC [American College of Cardiology]/AHA [American Heart Association] Guidelines) quality improvement initiative from January 2001 through March 2005. PATIENTS: 37,345 NSTE ACS patients younger than 65 years of age and 59,550 patients 65 years of age or older. MEASUREMENTS: Guideline-recommended treatments, and in-hospital outcomes. RESULTS: Medicaid was the primary payer for 18.7% (6999 of 37,345) of patients younger than age 65 years, whereas Medicare was the primary payer for 67.5% (40,199 of 59,550) of patients age 65 years or older. Medicaid patients were statistically significantly less likely to receive short-term (less than 24 hours) medications and to undergo invasive cardiac procedures than patients covered by HMO and private insurance. They also had higher mortality rates (2.9% vs. 1.2%; adjusted odds ratio, 1.33; 95% CI, 1.08 to 1.63). Medications and invasive procedures were used to a similar extent in patients with Medicare and HMO or private insurance, and respective mortality rates were not significantly different (6.2% vs. 5.6%; adjusted odds ratio, 1.08; 95% CI, 0.99 to 1.18). LIMITATIONS: Self-pay patients and patients without insurance were not assessed. CONCLUSIONS: NSTE ACS patients with Medicaid (but not Medicare) as the primary payer were less likely to receive evidence-based therapies and had worse outcomes than patients with HMO or private insurance as the primary payer. The causes of these treatment differences and solutions for narrowing the gaps in quality require further investigation. PMID- 17116919 TI - Pharmacogenomics: challenges and opportunities. AB - The outcome of drug therapy is often unpredictable, ranging from beneficial effects to lack of efficacy to serious adverse effects. Variations in single genes are 1 well-recognized cause of such unpredictability, defining the field of pharmacogenetics (see Glossary). Such variations may involve genes controlling drug metabolism, drug transport, disease susceptibility, or drug targets. The sequencing of the human genome and the cataloguing of variants across human genomes are the enabling resources for the nascent field of pharmacogenomics (see Glossary), which tests the idea that genomic variability underlies variability in drug responses. However, there are many challenges that must be overcome to apply rapidly accumulating genomic information to understand variable drug responses, including defining candidate genes and pathways; relating disease genes to drug response genes; precisely defining drug response phenotypes; and addressing analytic, ethical, and technological issues involved in generation and management of large drug response data sets. Overcoming these challenges holds the promise of improving new drug development and ultimately individualizing the selection of appropriate drugs and dosages for individual patients. PMID- 17116920 TI - Narrative review: the new epidemic of Clostridium difficile-associated enteric disease. AB - Antibiotic-associated diarrhea and colitis were well established soon after antibiotics became available. Early work implicated Staphylococcus aureus, but in 1978 Clostridium difficile became the established pathogen in the vast majority of cases. In the first 5 years (1978 through 1983), the most common cause was clindamycin, the standard diagnostic test was the cytotoxin assay, and standard management was to withdraw the implicated antibiotic and treat with oral vancomycin. Most patients responded well, but 25% relapsed when vancomycin was withdrawn. During the next 20 years (1983 through 2003), the most commonly implicated antibiotics were the cephalosporins, which reflected the rates of use; the enzyme immunoassay replaced the cytotoxin assay because of speed of results and technical ease of performance; and metronidazole replaced vancomycin as standard treatment, and principles of containment hospitals became infection control and antibiotic control. During the recent past (2003 to 2006), C. difficile has been more frequent, more severe, more refractory to standard therapy, and more likely to relapse. This pattern is widly distributed in the United States, Canada, and Europe and is now attributed to a new strain of C. difficile designated BI, NAP1, or ribotype 027 (which are synonymous terms). This strain appears more virulent, possibly because of production of large amounts of toxins, and fluoroquinolones are now major inducing agents along with cephalosporins, which presumably reflects newly acquired in vitro resistance and escalating rates of use. The recent experience does not change principles of management of the individual patient, but it does serve to emphasize the need for better diagnostics, early recognition, improved methods to manage severe disease and relapsing disease, and greater attention to infection control and antibiotic restraint. PMID- 17116921 TI - Update in pulmonary medicine. PMID- 17116922 TI - Changes in career decisions of internal medicine residents during training. AB - BACKGROUND: Little is known about the timing and stability of internal medicine resident career decisions during the course of residency training. OBJECTIVE: To assess changes in reported career plans among internal medicine trainees during their training. DESIGN: Observational cohort using data collected as part of the annual Internal Medicine In-Training Examination (IM-ITE) survey. SETTING: 411 internal medicine residency programs in North America participating in the annual IM-ITE. PARTICIPANTS: 2638 internal medicine residents who took the IM-ITE and responded to career plan questions on the test survey in all 3 years of training (2002-2004). MEASUREMENTS: Self-reported career plans for individual residents during their postgraduate year 1 (PGY-1), postgraduate year 2 (PGY-2), and postgraduate year 3 (PGY-3) of training. RESULTS: 2281 of 2638 residents (86.5%) identified a specific career plan in internal medicine during PGY-3. Of these 2281 residents, 1417 (62.1%) changed career plans at least once during the study period. Career plans reported by PGY-1 and PGY-2 residents matched subsequent PGY 3 plans for 55.1% and 68.4%, respectively. Six hundred eighty-six (26.0%) PGY-1, 278 (10.5%) PGY-2, and 205 (7.8%) PGY-3 residents remained undecided about their career plans at the time of the IM-ITE. Only 25.0% of graduating residents reported plans for a general internal medicine career. LIMITATIONS: The study cohort represents a convenience sample and is restricted to the subset of residents sitting for the IM-ITE and responding to career plan questions in all 3 years of postgraduate training. Career plans were assessed by self-report, and it is possible that residents who responded in all years of training differed from those who did not. CONCLUSIONS: In a subset of internal medicine residents in the class of 2004, career decisions changed late into residency training and enthusiasm for careers in general internal medicine remained low. PMID- 17116923 TI - Envisioning better approaches for dementia care. PMID- 17116924 TI - Career changes in medicine: Part II. PMID- 17116925 TI - Trials that matter: varenicline: a designer drug to help smokers quit. PMID- 17116926 TI - The genogram. PMID- 17116927 TI - Regional variations in health care intensity and physicians' perceptions of care quality. PMID- 17116928 TI - Withholding resuscitation: a new approach to prehospital end-of-life decisions. PMID- 17116929 TI - Low-molecular-weight heparin and bleeding: how do we lower risk but maintain benefit? PMID- 17116932 TI - Health Insurance Portability and Accountability Act Privacy Rule and research consent documents. PMID- 17116933 TI - Bisphosphonates and osteonecrosis of the jaw. PMID- 17116934 TI - Bisphosphonates and osteonecrosis of the jaw. PMID- 17116936 TI - Alterations in processes and priorities needed for new drug development. PMID- 17116937 TI - Estrogen receptor: methodology matters. PMID- 17116938 TI - Primary central nervous system lymphoma: the Memorial Sloan-Kettering Cancer Center prognostic model. AB - PURPOSE: The purpose of this study was to analyze prognostic factors for patients with newly diagnosed primary CNS lymphoma (PCNSL) in order to establish a predictive model that could be applied to the care of patients and the design of prospective clinical trials. PATIENTS AND METHODS: Three hundred thirty-eight consecutive patients with newly diagnosed PCNSL seen at Memorial Sloan-Kettering Cancer Center (MSKCC; New York, NY) between 1983 and 2003 were analyzed. Standard univariate and multivariate analyses were performed. In addition, a formal cut point analysis was used to determine the most statistically significant cut point for age. Recursive partitioning analysis (RPA) was used to create independent prognostic classes. An external validation set obtained from three prospective Radiation Therapy Oncology Group (RTOG) PCNSL clinical trials was used to test the RPA classification. RESULTS: Age and performance status were the only variables identified on standard multivariate analysis. Cut point analysis of age determined that patients age < or = 50 years had significantly improved outcome compared with older patients. RPA of 282 patients identified three distinct prognostic classes: class 1 (patients < 50 years), class 2 (patients > or =50; Karnofsky performance score [KPS] > or = 70) and class 3 (patients > or = 50; KPS < 70). These three classes significantly distinguished outcome with regard to both overall and failure-free survival. Analysis of the RTOG data set confirmed the validity of this classification. CONCLUSION The MSKCC prognostic score is a simple, statistically powerful model with universal applicability to patients with newly diagnosed PCNSL. We recommend that it be adopted for the management of newly diagnosed patients and incorporated into the design of prospective clinical trials. PMID- 17116939 TI - Is cytarabine useful in the treatment of acute promyelocytic leukemia? Results of a randomized trial from the European Acute Promyelocytic Leukemia Group. AB - PURPOSE: Several phase II studies have suggested that cytarabine (AraC) was not required in the treatment of newly diagnosed acute promyelocytic leukemia (APL) patients receiving all-trans-retinoic acid (ATRA), an anthracycline, and maintenance therapy, and we aimed at confirming this finding in a randomized trial. PATIENTS AND METHODS: Newly diagnosed APL patients younger than age 60 years with a WBC count of less than 10,000/microL were randomly assigned to receive either ATRA combined with and followed by three daunorubicin (DNR) plus AraC courses and a 2-year maintenance regimen (AraC group) or the same treatment but without AraC (no AraC group). Patients older than age 60 years and patients with initial WBC count of more than 10,000/microL were not randomly assigned but received risk-adapted treatment, with higher dose of AraC and CNS prophylaxis in patients with WBC counts more than 10,000/microL. RESULTS: Overall, 328 (96.5%) of 340 patients achieved complete remission (CR). In the AraC and the no AraC groups, the CR rates were 99% and 94% (P = .12), the 2-year cumulative incidence of relapse (CIR) rates were 4.7% and 15.9% (P = .011), the event-free survival (EFS) rates were 93.3% and 77.2% (P = .0021), and survival rates were 97.9% and 89.6% (P = .0066), respectively. In patients younger than age 60 years with WBC counts more than 10,000/microL, the CR, 2-year CIR, EFS, and survival rates were 97.3%, 2.9%, 89%, and 91.9%, respectively. CONCLUSION: These results support a role for AraC in addition to ATRA and anthracyclines in the treatment of newly diagnosed APL, at least using DNR at the cumulative dose we used and with the consolidation and maintenance regimens we used. PMID- 17116940 TI - Allogeneic marrow stem-cell transplantation from human leukocyte antigen identical siblings versus human leukocyte antigen-allelic-matched unrelated donors (10/10) in patients with standard-risk hematologic malignancy: a prospective study from the French Society of Bone Marrow Transplantation and Cell Therapy. AB - PURPOSE: To investigate the influence of donor type (human leukocyte antigen [HLA] -identical sibling donor versus HLA-A-, HLA-B-, HLA-Cw-, HLA-DRB1-, and HLA DQB1-identical unrelated donors, or so-called 10/10) on the outcome of patients who underwent allogeneic stem-cell transplantation (alloSCT), adjusting for other prognostic factors, in patients with standard-risk hematologic malignancy. PATIENTS AND METHODS: Between March 2000 and January 2003, we prospectively investigated the outcome of 236 consecutive patients with standard-risk malignancy from 12 French centers. Fifty-five patients underwent alloSCT from an unrelated HLA-identical donor at the allelic level, whereas 181 patients received an alloSCT from an HLA-identical sibling. Diagnoses included acute leukemia (n = 175), chronic myeloid leukemia (n = 43), and myelodysplastic syndrome (MDS; n = 18). All patients received unmodified marrow graft following myeloablative conditioning with cyclophosphamide and total-body irradiation. Graft-versus-host disease (GVHD) prophylaxis consisted of cyclosporine and short-course methotrexate in all patients. RESULTS: In multivariable analysis, overall survival and transplantation-related mortality were adversely influenced by recipient cytomegalovirus (CMV) -positive serology, age of donor older than 37 years, and the occurrence of acute grade > or = II GVHD. Event-free survival rates were lower for patients with recipient CMV-positive serology. Acute grades II to IV GVHD rates were higher for patients with chronic myeloid leukemia (CML). No factor was found to influence either relapse or acute grades III to IV GVHD. The effect of donor type was nonsignificant for all criteria. CONCLUSION: In patients with standard-risk malignancy, transplantation from unrelated HLA allellically matched donors led to outcomes similar to those from HLA-identical sibling donors. PMID- 17116941 TI - Sequential adjuvant epirubicin-based and docetaxel chemotherapy for node-positive breast cancer patients: the FNCLCC PACS 01 Trial. AB - PURPOSE: The PACS 01 trial compared six cycles of fluorouracil, epirubicin, and cyclophosphamide (FEC) with a sequential regimen of three cycles of FEC followed by three cycles of docetaxel (FEC-D) as adjuvant treatment for women with node positive early breast cancer. PATIENTS AND METHODS: Between June 1997 and March 2000, 1,999 patients with operable node-positive breast cancer were randomly assigned to either FEC every 21 days for six cycles, or three cycles of FEC followed by three cycles of docetaxel, both given every 21 days. Hormone-receptor positive patients received tamoxifen for 5 years after chemotherapy. The primary end point was 5-year disease-free survival (DFS). RESULTS: Median follow-up was 60 months. Five-year DFS rates were 73.2% with FEC and 78.4% with FEC-D (unadjusted P = .011; adjusted P = .012). Multivariate analysis adjusted for prognostic factors showed an 18% reduction in the relative risk of relapse with FEC-D. Five-year overall survival rates were 86.7% with FEC and 90.7% with FEC-D, demonstrating a 27% reduction in the relative risk of death (unadjusted P = .014; adjusted P = .017). The incidence of grade 3 to 4 neutropenia, the need for hematopoietic growth factor, and incidence of nausea/vomiting were higher with FEC. Docetaxel was associated with more febrile neutropenia in the fourth cycle, stomatitis, edema, and nail disorders. Though rare overall, there were fewer cardiac events after FEC-D (P = .03), attributable mainly to the lower anthracycline cumulative dose. CONCLUSION: Sequential adjuvant chemotherapy with FEC followed by docetaxel significantly improves disease-free and overall survival in node-positive breast cancer patients and has a favorable safety profile. PMID- 17116942 TI - Locoregional relapse and distant metastasis in conservatively managed triple negative early-stage breast cancer. AB - PURPOSE: To determine the prognostic significance of triple negative breast cancers with respect to locoregional relapse and distant metastasis in conservatively managed breast cancer patients. PATIENTS AND METHODS: A database of conservative managed (conservative surgery followed by radiation) patients, in whom all three markers (estrogen receptor, progesterone receptor, and HER2/neu) were available, was reviewed. Patients were classified as triple negative if they tested negative for all three markers. Of 482 patients with all three markers available, 117 were classified as triple negative. RESULTS: As of September 2005, with a median follow-up time of 7.9 years, of the 482 patients in the study, there have been 53 in-breast relapses, 10 nodal relapses, 77 distant relapses, and 69 deaths. At 5 years, the triple negative cohort had a poorer distant metastasis-free rate compared with the other subtypes (67% v 82%, respectively; P = .002). Triple negative subtype was an independent predictor of distant metastasis (hazard ratio = 2.14; 95% CI, 1.31 to 3.53; P = .002) and cause specific survival (hazard ratio = 1.79; 95% CI, 1.03 to 3.22; P = .047). There was no significant difference in local control between the triple negative and other subtypes (83% v 83%, respectively). Of 99 BRCA-tested patients in this cohort, 10 had deleterious mutations in BRCA1, and seven had mutations in BRCA2. Of 10 BRCA1 patients, eight were triple negative, whereas only one of seven BRCA2 patients was triple negative (P < .001). CONCLUSION: Patients classified as triple negative have a poor prognosis. However, there was no evidence that these patients are at higher risk for local relapse after conservative surgery and radiation. Patients with BRCA1 mutations develop predominantly triple negative tumors. PMID- 17116943 TI - Polymorphisms in XRCC1, XRCC3, and CCND1 and survival after treatment for metastatic breast cancer. AB - PURPOSE: Single nucleotide polymorphisms (SNPs) in DNA repair and cell cycle control genes may alter protein function and therefore the efficacy of DNA damaging chemotherapy. We retrospectively evaluated the association of SNPs in DNA repair genes, XRCC1-01 (Arg399Gln) and XRCC3-01 (Thr241Met), and a cell cycle control gene, CCND1-02 (A870G), with progression-free survival (PFS) and breast cancer specific survival (BCSS) in patients with metastatic breast cancer (MBC). PATIENTS AND METHODS: SNPs in 95 patients with MBC enrolled onto one of five prospective clinical trials of high-dose chemotherapy and autologous stem-cell transplantation were evaluated using genotyping assays. RESULTS: For XRCC1-01, the hazard ratio (HR) for BCSS was 2.8 (95% CI, 1.60 to 5.00) and the HR for PFS was 2.0 (95%CI, 1.12 to 3.43). For XRCC3-01, the HR for BCSS was 2.0 (95%CI, 1.12 to 3.70) and the HR for PFS was 2.0 (95%CI, 1.09 to 3.59). For CCND1-02, the HR for BCSS was 1.8 (95%CI, 1.12 to 2.78) and the HR for PFS was 1.8 (95%CI, 1.15 to 2.85). Patients carrying one variant genotype (HR, 1.7; 95%CI, 1.07 to 2.82) or combinations of any two variant genotypes (HR, 4.7; 95% CI, 2.41 to 8.94) had significantly poorer BCSS compared with patients carrying zero variants. In multivariable analysis, XRCC1-01, presence of liver metastases, and bone metastases independently predicted BCSS. Combinations of any two variant genotypes were stronger independent predictors of BCSS and PFS than the presence of liver or bone metastases. CONCLUSION: XRCC1-01, XRCC3-01, and CCND1-01 may be predictive of survival outcome in patients with MBC treated with DNA damaging chemotherapy. PMID- 17116944 TI - Immunohistochemical detection using the new rabbit monoclonal antibody SP1 of estrogen receptor in breast cancer is superior to mouse monoclonal antibody 1D5 in predicting survival. AB - PURPOSE: Estrogen receptor (ER) expression predicts improved breast cancer specific survival and reduced risk of recurrence and is targeted in breast cancer therapy. A high-quality antibody to identify ER-positive patients plays an important role in clinical decision making for women with breast cancer. This study evaluates immunohistochemistry using two anti-ER antibodies, a new rabbit monoclonal antibody (SP1) and the mouse monoclonal antibody (1D5), in relation to biochemical ER assay results and clinical data on survival and adjuvant systemic therapy. PATIENTS AND METHODS: A population-based tissue microarray series of 4,150 invasive breast cancers was constructed. All patients had staging, pathology, treatment, and follow-up information. The median follow-up was 12.4 years and the median age at diagnosis 60 years. Survival analysis and log-rank tests were used to evaluate the prognostic value of ER status and correlations with clinical data. RESULTS: Among the 4,105 samples interpretable for both antibodies, SP1 detected ER positivity in 69.5% and 1D5 in 63.1% of cases. Both monoclonal antibodies are demonstrated to be good prognostic indictors for breast cancer-specific and relapse-free survival. In multivariate analysis, including age, tumor size, grade, and lymphovascular and nodal status, SP1 was a better independent prognostic factor than 1D5. Among patients with discrepant ER results, the 8% of patients who were SP1 positive/1D5 negative showed good outcomes, and the 2% SP1-negative/1D5 positive had poor outcomes. Maintaining the same 92% specificity and 98% positive predictive value, SP1 is 8% more sensitive than 1D5 using biochemical assay as gold standard. CONCLUSION: SP1 represents an improved standard for ER immunohistochemistry assessment in breast cancer. PMID- 17116945 TI - Psychiatric and emotional sequelae of surgical amputation. AB - The prevalence of posttraumatic stress symptoms after the experience of amputation is not well established. The current study gathered data on the prevalence of posttraumatic stress disorder (PTSD) and other psychiatric disorders after amputation. Participants were recruited from a large Northeastern rehabilitation hospital and were assessed with structured clinical interviews. The data suggest that planned surgical amputations resulting from chronic illness do not frequently lead to PTSD symptoms. In contrast, data suggest that amputation resulting from accidental injury may lead to a higher prevalence of PTSD, in part because of the emotional stress surrounding the accident. PMID- 17116946 TI - Measuring somatic symptoms with the CES-D to assess depression in cancer patients after treatment: comparison among patients with oral/oropharyngeal, gynecological, colorectal, and breast cancer. AB - There is a high prevalence of depression after cancer treatment. In the literature, several authors have raised questions about assessing somatic symptoms to explore depression after cancer treatment. These somatic sequelae are a consequence of cancer treatment and should cause higher depression rates in cancer patients. In this study, the Somatic domain on a depression questionnaire, the Center for Epidemiologic Studies-Depression scale (CES-D) was analyzed in different cancer patients after treatment, as compared with a control group. Data from 566 cancer patients (oral/oropharyngeal, gynecological, colorectal, and breast cancer) and 255 randomly chosen comparison patients were analyzed. The total score on the CES-D domain of Somatic Retarded Activity significantly differed between the cancer and comparison groups; but the cancer groups showed both less somatic morbidity (colorectal cancer) and more somatic morbidity (oral/oropharyngeal, breast) than the comparison group. In the analyses of the CES-D with and without the Somatic domain, the prevalence of depression symptoms with the Somatic domain is lower for the cancer groups. Authors conclude that cancer patients are not a homogenous group as regards somatic sequelae. Evidence for removing Somatic items from the CES-D for patients after cancer treatment was not confirmed. PMID- 17116947 TI - Phenomenology of delirium in cancer patients, as described by the Memorial Delirium Assessment Scale (MDAS) and the Delirium Rating Scale (DRS). AB - This study was based on the data collected on a consecutive sample of 106 cancer patients referred for mental status evaluation. All patients were evaluated by use of the Confusion Assessment Method (CAM) algorithm, the Delirium Rating Scale (DRS), the Memorial Delirium Assessment Scale (MDAS), and a question about the subjective perception of delirium. After comparing the diagnostic criteria of delirium on the DSM-III-R and DSM-IV, authors evaluated the ability of all DRS and MDAS items to discriminate delirium versus non-delirium patients, testing the difference in the distribution of the individual MDAS and DRS item scores. Authors also assessed the relationship between delirium diagnosis and the subjective perception of delirium. The MDAS showed a greater number of discriminating items. The items that proved to be less discriminating were "Hallucinations" and "Lability of Mood" on the DRS. Subjective perception only partially discriminated delirium from non-delirium patients. The way in which the DRS and MDAS reflected the DSM criteria are therefore partially different. PMID- 17116948 TI - Prevalence and correlates of major depressive disorder in Nigerian outpatients with heart failure. AB - This study aims to estimate the prevalence and correlates of major depressive disorder (MDD) in Nigerian outpatients with heart failure. Authors assessed patients with heart failure (N = 102) for DSM-IV diagnosis of MDD and obtained sociodemographic and clinical data. MDD was found in 28 (27.5%) of the patients. The significant correlates predicting MDD included unemployment and disability due to the illness, more severe illness (NYHA class), age younger than 60 years, and not being married. These factors should be considered in planning further studies and in screening and intervention programs for patients with heart failure. PMID- 17116949 TI - Naked patients in the general hospital: differential diagnosis and management strategies. AB - Physicians and patients are frequently concerned, and, at times, distressed, by nakedness during clinical encounters. When nakedness appears, clinicians should attempt to establish the reason for it and determine whether it is appropriate for the situation. Establishing the etiology of nudity can facilitate care by hospital staff and help to modulate their countertransference reactions and behavior. The authors present and discuss three cases involving nudity at times other than during the physical examination, within the context of differential diagnosis and treatment alternatives. PMID- 17116950 TI - The prevalence of the metabolic syndrome in psychiatric inpatients with primary psychotic and mood disorders. AB - Patients with severe mental illness have elevated rates of cardiovascular disease (CVD) and diabetes compared with the general population, but little is known about the prevalence of the metabolic syndrome that predisposes patients with severe mental illness to both medical conditions. The purpose of this study was to assess the prevalence of the metabolic syndrome by surveying hospital records of psychiatric inpatients with severe mood and psychotic disorders. The study group was 102 consecutively admitted adult patients with a primary DSM-IV diagnosis of a mood or psychotic disorder. Criteria for comorbid metabolic syndrome required at least three of the five factors defined by the National Cholesterol Education Program. The prevalence of the metabolic syndrome was 38.6% in this cohort, and it was associated with increasing age, body mass index, and Caucasian ethnicity. The metabolic syndrome was common in this cohort of psychiatric inpatients, and the high rate of the metabolic syndrome likely represents an intermediate step in the future development of CVD and diabetes, which may provide a point of early intervention to prevent the occurrence of these two medical illnesses in chronically mentally ill patients. PMID- 17116951 TI - Somatoform disorder in primary care: course and the need for cognitive-behavioral treatment. AB - Medically unexplained physical symptoms are prevalent in primary care. Of all patients attending the family physician, 16% have a somatoform disorder as described by DSM-IV. Cognitive-behavioral treatment has been demonstrated to be effective in secondary care. However, the course of somatoform disorders and their need for treatment have not yet been established in primary care. In this study, data from 1,046 attendees in family practice were analyzed for prevalence, course, and eligibility for treatment. Over a 6-month follow-up, the prevalence of somatoform disorder decreased from 16.1% to 12.3%. After assessment of eligibility, about 5% of patients demonstrated a need for treatment. PMID- 17116952 TI - Desire for hastened death among patients with advanced AIDS. AB - The recent debate over legalization of physician-assisted suicide has fueled interest in understanding factors that lead medically ill patients to seek a hastened death. The authors investigated the prevalence and predictors of desire for hastened death in 372 patients with advanced AIDS who were newly admitted to a palliative-care facility. Clinician-rated and self-report measures of desire for hastened death, depression, hopelessness, spiritual well-being, social support, pain, and physical symptom burden were administered to assess the factors that correspond to a high desire for death. The prevalence ranged from 4.6% to 8.3%, significantly lower than in previous studies of patients with advanced or terminal cancer. Multivariate models revealed significant and unique effects for both hopelessness and depression, with these variables accounting for a large proportion of the variance in each model. Authors discuss the implications of these findings for palliative care practice and the assisted suicide debate. PMID- 17116953 TI - The triple threat for chronic disease: obesity, race, and depression. AB - The authors investigated the interrelationships between race, obesity, depression, and chronic disease by abstracting data from all primary-care patients seen at a family-medicine clinic over a 3-year period. A total of 8,197 patients were included in the analysis. Sixty-three percent of patients were either overweight (26%) or obese (37%). African-American race, obesity, and having a diagnosis of depression each independently and significantly increased the likelihood of having a chronic disease. Also, these risk factors interacted to create an increased likelihood of disease prevalence. Thus, obesity, race, and depression interacted to create a "triple threat" of developing certain chronic diseases. PMID- 17116955 TI - The role of psychosomatic-medicine psychiatrists in bioethics: a survey study of members of the academy of psychosomatic medicine. AB - The Bioethics Subcommittee of The Academy of Psychosomatic Medicine developed a survey to assess the involvement of psychosomatic-medicine psychiatrists in bioethics and the extent of their participation on bioethics committees and in the teaching of bioethics. Of 599 Academy members surveyed, 122 (20.4%) responded. The majority of respondents reported that the management of bioethical dilemmas had a significant impact on their work in psychosomatic medicine. Many respondents were involved in teaching bioethics and in serving on ethics committees. The majority of respondents reported psychiatry-resident involvement on ethics committees. Bioethics work is an integral part of the fabric of psychosomatic medicine. PMID- 17116956 TI - Fatal familial insomnia. PMID- 17116957 TI - Hyperthyroidism associated with topiramate treatment. PMID- 17116958 TI - Baking-soda pica in an adolescent patient. PMID- 17116959 TI - Cardioversion of persistent atrial arrhythmia after treatment with venlafaxine in successful management of major depression and posttraumatic stress disorder. AB - There is increasing evidence linking depression and cardiovascular disease. However, the authors could find no literature directly linking depression with atrial fibrillation or atrial flutter. The authors report the case of a patient with uncontrolled atrial arrhythmia who cardioverted to normal sinus rhythm after treatment of major depressive disorder (MDD) and posttraumatic stress disorder (PTSD) with venlafaxine. The authors discuss comorbidity of MDD and atrial fibrillation, and explore evidence of venlafaxine as an antiarrhythmic agent. Further research is needed to establish the clinical role of venlafaxine as a Class 1 antiarrhythmic agent and any association between atrial arrhythmias and MDD and PTSD. PMID- 17116960 TI - Visual hallucinations and metoprolol. PMID- 17116961 TI - Neurosyphilis presenting as rhabdomyolysis and acute renal failure with subsequent irreversible psychosis and dementia. PMID- 17116962 TI - Ciprofloxacin-induced manic episode. PMID- 17116963 TI - Ethical impasses in the care of patients with hepatitis C. PMID- 17116966 TI - Airway epithelial (nasal) cell monolayers used to study Pseudomonas aeruginosa invasion are hyperpolarized and not representative of the human airway epithelium. PMID- 17116967 TI - Evaluation of antimycobacterial and DNA gyrase inhibition of fluoroquinolone derivatives. AB - The antimycobacterial activity (both in vitro and in vivo) and DNA gyrase inhibition of newly synthesized fluoroquinolone derivatives were tested against Mycobacterium tuberculosis H(37)Rv and Mycobacterium smegmatis, respectively. Among the synthesized compounds, compound F11 was found to exhibit the most potent in vitro antimycobacterial activity with a MIC value of 0.78 microg/ml, and a selectivity index of more than 80 while not being cytotoxic to the Vero cell line up to 62.5 microg/ml. When evaluated for in vivo antimycobacterial activity, compound F11 demonstrated a paramount decrease of bacterial load in lung and spleen tissues compared to the control and better than the standard drug ciprofloxacin. PMID- 17116968 TI - Magenta pigment produced by fungus. AB - A fungus producing magenta was isolated from cellulosic material by visual observation on Czapek's agar media and the product was conventionally analyzed. The fungal strain that produced magenta pigment was closely related to Phoma herbarum. The type of fibers added to Czapek's medium influenced which pigments were produced. Mycelia attached to the surface of nylon-6 and excreted magenta pigment into the fibers. The pigment structure was partially determined. This is the first report of the production of magenta pigment by a microorganism specifically in the presence of nylon-6 fibers, via an unknown mechanism. This phenomenon raises the question of why and how the fungus disperses the pigment inside the fiber and suggests that fabrics can be dyed using microorganisms. PMID- 17116969 TI - Studies on poly-beta-hydroxybutyrate synthase activity of Nostoc muscorum. AB - This study compares the PHB synthase activity of Nostoc muscorum, a N(2)-fixing cyanobacterium under control (grown in usual BG-11 medium), nitrogen (N) and phosphorus (P) deprivation and chemoheterotrophic conditions. Specific activity of PHB synthase did not depict significant variations in the latter three types of cultures, except for the control one, where a significantly lower activity was recorded. PHB synthase activity was detected only in the soluble fractions of both the control as well as cells incubated under chemoheterotrophic conditions. A K(m) of 80.2 microM DL-beta-hydroxybutyryl-CoA and V(max) of 197.5 nmol thiobenzoate (TNB) mg protein(-1)min(-1) were observed for the enzyme. PHB synthase remained insensitive to acetyl-CoA, ATP, NADP, NADPH supplementation under in vitro condition. Addition of acetyl phosphate was found to activate the enzyme and the level of activation was dependent on the concentration of acetyl phosphate supplementation. Inhibition of PHB synthase in 2,3-butanedione supplemented cultures and reactivation following acetyl phosphate addition proved the post-translational control of acetyl phosphate over PHB synthase. PMID- 17116970 TI - Biosurfactant production by Pseudomonas aeruginosa A41 using palm oil as carbon source. AB - Biosurfactant production by Pseudomonas aeruginosa A41, a strain isolated from seawater in the gulf of Thailand, was examined when grown in defined medium containing 2% vegetable oil or fatty acid as a carbon source in the presence of vitamins, trace elements and 0.4% NH(4)NO(3), at pH 7 and 30 degrees C with 200 rpm-shaking for 7 days. The yield of biosurfactant steadily increased even after a stationary phase. Under such conditions the surface tension of the medium was lowered from 55-70 mN/m to 27.8-30 mN/m with every carbon source tested. However, types of carbon sources were found to affect biosurfactant yield. The yields of rhamnolipid biosurfactant were 6.58 g/L, 2.91 g/L and 2.93 g/L determined as rhamnose content when olive oil, palm oil and coconut oil, respectively, were used as a carbon source. Among them, biosurfactant obtained from palm oil was the best in lowering surface tension of the medium. Increase in biosurfactant activities in terms of oil displacement test and rhamnose content were observed to be higher with shorter chain fatty acids than that of the longer chains (C12>C14>C16). In addition, we found that C18:2, highly unsaturated fatty acid, showed higher oil displacement activity and rhamnose content than that of C18:1. The optimal oil displacement activity was found at pH 7-9 and in the presence of 0.5-3% NaCl. The oil displacement activity was stable to temperatures up to 100 degrees C for 15 h. Surface tension reduction activity was relatively stable at pH 2-12 and 0-5% of NaCl. Emusification activity tested with various types of hydrocarbons and vegetable oils showed similarity of up to 60% stability. The partially purified biosurfactant via TLC and silica gel column chromatography gave three main peaks on HPLC with mass spectra of 527, 272, and 661 m/z respectively, corresponding to sodium-monorhamnodecanoate, hydroxyhexadecanoic acid and an unknown compound, respectively. PMID- 17116971 TI - Geobacillus toebii subsp. decanicus subsp. nov., a hydrocarbon-degrading, heavy metal resistant bacterium from hot compost. AB - A thermophilic, spore-forming bacterial strain L1(T) was isolated from hot compost "Pomigliano Environment" s.p.a., Pomigliano, Naples, Italy. The strain was identified by using a polyphasic taxonomic approach. L1(T) resulted in an aerobic, gram-positive, rod-shaped, thermophilic with an optimum growth temperature of 68 degrees C chemorganotrophic bacterium which grew on hydrocarbons as unique carbon and energy sources and was resistant to heavy metals. The G+C DNA content was 43.5 mol%. Phylogenetic analysis of 16S rRNA gene sequence and Random Amplified Polymorphic DNA-PCR (RAPD-PCR) analysis of L1(T) and related strains showed that it forms within Geobacillus toebii, a separate cluster in the Geobacillus genus. The composition of cellular fatty acids analyses by Gas-Mass Spectroscopy differed from that typical for the genus Geobacillus in that it is lacking in iso-C15 fatty acid, while iso-C16 and iso C17 were predominant. Isolates grew on a rich complex medium at temperatures between 55-75 degrees C and presented a doubling time (t(d)) of 2 h and 6 h using complex media and hydrocarbon media, respectively. Among hydrocarbons tested, n decane (2%) was the more effective to support the growth (1 g/L of wet cells). The microorganism showed resistance to heavy metal tested during the growth. Furthermore, intracellular alpha-galactosidase and alpha-glucosidase enzymatic activities were detectable in the L1(T) strain. Based on phenotypic, phylogenetic, fatty acid analysis and results from DNA-DNA hybridization, we propose assigning a novel subspecies of Geobacillus toebii, to be named Geobacillus toebii subsp. decanicus subsp. nov., with the type strain L1(T) (=DSM 17041=ATCC BAA 1004). PMID- 17116972 TI - Cellular polyamines of lower eukaryotes belonging to the phyla Glaucophyta, Rhodophyta, Cryptophyta, Haptophyta and Percolozoa. PMID- 17116973 TI - Identification of strains assigned to the genus Asaia Yamada et al. 2000 based on 16S rDNA restriction analysis. PMID- 17116974 TI - An experimental model of prolonged esophagitis with sphincter failure in the rat and the therapeutic potential of gastric pentadecapeptide BPC 157. AB - We report a simple novel rat model that combines prolonged esophagitis and parallel sphincters failure. The anti-ulcer gastric pentadecapeptide BPC 157, which was found to be stable in gastric juice, and is being evaluated in inflammatory bowel disease trials, is an anti-esophagitis therapy that recovers failed sphincters. Twelve or twenty months after the initial challenge (tubes sutured into sphincters for one week and then spontaneously removed by peristalsis), rats exhibit prolonged esophagitis (confluent hemorrhagic and yellowish lesions, thinner epithelium and superficial corneal layer, with stratification derangement); constantly lowered pressure of both sphincters (assessed by using a water manometer connected to the drainage port of a Foley catheter implanted into the stomach either through esophageal or duodenal incision); and both lower esophageal and pyloric sphincter failure. Throughout the esophagitis experiment, BPC 157 was given at either 10 micro g/kg, i.p., once a day (last application 24 h before assessment) or alternatively, it was given continuously in drinking water at 0.16 micro g/ml (12 ml/rat). This treatment recovers i) esophagitis (macroscopically and microscopically, at either region or investigated time period) and ii) pressure in both sphincters (cmH2O). In addition, BPC 157 (10 micro g/kg) or saline (1 ml/rat, 5 ml/kg) was specifically given directly into the stomach; pressure assessment was performed at 5 min thereafter. The effect of BPC 157 is specific because in normal rats, it increases lower esophageal sphincter-pressure, but decreases pyloric sphincter pressure. Ranitidine, given as the standard drug using the same protocol (50 mg/kg, i.p., once daily; 0.83 mg/ml in drinking water; or 50 mg/kg directly into the stomach) had no effect. PMID- 17116975 TI - Fibroblast growth factor-2 suppresses oridonin-induced L929 apoptosis through extracellular signal-regulated kinase-dependent and phosphatidylinositol 3-kinase independent pathway. AB - Oridonin, isolated from Rabdosia rubescences, has been reported to exert cytotoxic effects on L929 cells. In this study, we investigated the mechanisms of FGF-2 protection of L929 cells from oridonin-induced apoptosis. Phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB) signal did not mediate this effect because the PI3K inhibitor wortmannin failed to reverse this protection and PKB activation was not observed in this process. In contrast, the extracellular signal-regulated kinase (ERK) was responsible for this rescue because its inhibition abolished the protective effect of fibroblast growth factor (FGF)-2. ERK had dual regulatory functions: mediating cell apoptosis or preventing cells from initiating the apoptotic response by phosphorylation or promoting expression of Bcl-2 in dependence of different stimuli. In L929 cells treated with oridonin alone, the activated ERK decreased the ratio of Bcl-2/Bax by mediating the phosphorylation of Bcl-2, resulting in apoptosis; the Ras inhibitor manumycin A and Raf inhibitor GW5074 failed to inhibit this apoptosis, indicating that there is a signal other than Ras/Raf pathway activated ERK. However, in the presence of FGF-2, Bcl-2 phosphorylation was blocked, and the Ras/Raf/ERK signal pathway was activated and protected against the oridonin induced apoptosis by the alternative function of promoting of Bcl-2 expression. PMID- 17116976 TI - Pharmacokinetics and anti-asthmatic potential of non-parenterally administered recombinant human interleukin-1 receptor antagonist in animal models. AB - The objectives of this study were to define the pharmacokinetics of recombinant human interleukin-1 receptor antagonist (rhIL-1ra) and its effects on allergic asthma, cell adhesion molecules, and upper respiratory tract following non parenteral administration in animals. Pharmacokinetics and immunomodulating effects of rhIL-1ra were investigated in Sprague-Dawley rats and asthmatic guinea pigs, respectively. Effects on the upper respiratory tract following the applications of rhIL-1ra were investigated on the ex vivo nasal mucosa of Sprague Dawley rats and in situ in the upper palate of Chinese toads. Absolute bioavailabilities after intratracheal and intranasal administrations of rhIL-1ra were 94.3% and 24.8%, respectively. After administration of rhIL-1ra solution as ultrasonic spraying, the asthmatic symptom in guinea pigs was obviously attenuated. The plasma soluble intercellular cell adhesion molecule (sICAM-1) and P-selectin levels in asthmatic guinea pigs were each dose-dependently reduced with the increase of rhIL-1ra dose. The rhIL-1ra solution after administration via the airway seemed to have no impact on the integrity of nasal mucosa and mucocilia clearance in the upper respiratory tract. The present study provides evidence that rhIL-1ra effectively suppresses allergen-induced asthmatic symptoms through spraying, which corresponds to nasal and pulmonary absorption or both, and the efficacy is associated with downregulation of sICAM-1 and P-selectin expressions. PMID- 17116977 TI - Type of electrical stimulation influences diaphragm response to adrenoceptor and calcium channel modulators: the role of extracellular and intracellular calcium events. AB - The influence of direct single pulse and subtetanic electrical stimulation (ES) on isolated rat hemidiaphragm response to adrenoceptor antagonists and calcium channel blockers was investigated. Muscle contractility was stimulated by cumulative micromolar noradrenaline. Noradrenaline effects on developed tension (Td) in the presence of various alpha- and beta-adrenoceptor antagonists (e.g., prazosin and ICI 118551) were qualitatively different during different types of ES. Also, intact L-voltage calcium channels were necessary for noradrenaline induced potentiation of Td during both types of ES, while the balance between ryanodine receptors- and inositol triphosphate (IP3)-related calcium events in the muscle was influenced by the pattern of the ES. PMID- 17116978 TI - Proteolytic activation of Vibrio mimicus (Vm) major outer membrane protein haemagglutinin (HA) with Vm-HA/protease: Implication for understanding bacterial adherence. AB - Vibrio mimicus (Vm) haemagglutinins (HAs), such as an extracellular HA/protease (Vm-HA/protease) and a major outer membrane protein-HA (Vm-OMPHA), have been recognized as the putative adherence factors for the bacterium. However, the mechanism by which HAs coordinate the adherence function of the bacterium remains as yet unknown. We report herein the positive interaction between Vm-HA/protease and Vm-OMPHA resulting in significant enhancement of the haemagglutinating ability. In this interaction, no cleaved polypeptide was detected; however, limited proteolysis of Vm-OMPHA was confirmed by SDS-PAGE. The proteolytic activation of the native cell-associated Vm-OMPHA by limited proteolysis was also demonstrated in several V. mimicus strains. Proteolytic activation of OMPHA was also achieved with various proteases from bacterial and eukaryotic sources. These findings may indicate a novel coordination of V. mimicus HAs in the adherence of the bacterium. PMID- 17116979 TI - Detection and quantification of specific IgE antibodies against eight Malassezia species in sera of patients with atopic dermatitis by using an enzyme-linked immunosorbent assay. AB - The lipophilic yeast Malassezia, a member of the cutaneous microflora, is an exacerbating factor in atopic dermatitis (AD). Of the 11 currently recognized species, M. globosa and M. restricta are found to frequently colonize the skin of AD patients. In this study, we attempted to quantify specific IgE antibodies against eight Malassezia species, namely, M. dermatitis, M. furfur, M. globosa, M. obtusa, M. pachydermatis, M. slooffiae, M. sympodialis, and M. restricta, in sera from AD patients by using an enzyme-linked immunosorbent assay (ELISA). The specific IgE value against M. restricta was greater than those against other Malassezia species. Competitive ELISA inhibition tests revealed that M. restricta contained species specific as well as shared antigens. Therefore, M. restricta could be considered as a candidate diagnostic antigen for detecting anti Malassezia IgE in sera from AD patients. PMID- 17116980 TI - New epitope peptides derived from hepatitis C virus (HCV) 2a which have the capacity to induce cytotoxic T lymphocytes in HLA-A2+ HCV-infected patients. AB - Because cytotoxic T lymphocytes (CTLs) play an important role in the specific immunotherapy of hepatitis C virus (HCV) infection, a series of CTL epitopes has been defined from HCV genotype 1a or 1b protein. Here, we attempted to identify HCV2a-derived epitopes that are capable of inducing HLA-A2-restricted and peptide specific CTLs. Peripheral blood mononuclear cells (PBMCs) of HLA-A2+ HCV2ainfected patients or healthy donors were stimulated in vitro with each of the HCV2a-derived peptides, which were prepared based on the HLA-A2-binding motif, and their peptide-specific and HLA-A2-restricted cytotoxicities were examined. The HCV2a 432-441, HCV2a 716-724, and HCV2a 2251-2260 peptides were found to efficiently induce peptide-specific CTLs from the PBMCs of HLA-A2+ HCV2ainfected patients. Cytotoxicity was mainly mediated by CD8+ T cells in a HLA class I-restricted manner. These results indicate that the HCV2a 432-441, HCV2a 716-724, and HCV2a 2251-2260 peptides might be applicable for peptide-based immunotherapy of HLA-A2+ HCV2a-infected patients. PMID- 17116981 TI - Monitoring and survival of Lactobacillus gasseri SBT2055 in the human intestinal tract. AB - The monitoring and survival of Lactobacillus gasseri SBT2055 in the human intestinal tract was investigated with seven healthy subjects having a low number of fecal lactobacilli. An increase of fecal lactobacilli (10(3.2-5.2) CFU/g feces) was recognized after ingestion of yogurt with SBT2055 by the subjects. A high positive rate of L. gasseri in fecal lactobacilli detected from the subjects (over 70% at 2nd weeks of feeding) was also observed during the ingestion period using the species-specific PCR system. These findings indicate that the SBT2055 strain in yogurt survived in the human intestinal tract and was recovered from human feces. PMID- 17116982 TI - Involvement of the Vibrio parahaemolyticus pvsC gene in export of the siderophore vibrioferrin. AB - The pvsC gene of unknown function has been found in the iron-regulated vibrioferrin biosynthesis operon of Vibrio parahaemolyticus (Tanabe, T. et al., J. Bacteriol. 185: 6938-6949, 2003). The amino acid sequence deduced from the gene showed significant similarity to 12-transmembrane segment efflux pumps belonging to the major facilitator superfamily. A nonpolar deletion of pvsC in V. parahaemolyticus resulted in a reduced release of vibrioferrin into the medium. Vibrioferrin release could be regained by introducing the intact pvsC gene on a complementing plasmid. These results indicate that the pvsC gene product functions as an inner membrane exporter of vibrioferrin. PMID- 17116983 TI - Human milk glycoproteins inhibit the adherence of Salmonella typhimurium to HeLa cells. AB - The ability of human milk, as well as its protein fractions, to inhibit the adhesion and invasion of Salmonella typhimurium to HeLa cells was investigated. The results revealed that milk secretory immunoglobulin A (sIgA) inhibited neither the adherence nor the bacterial invasion; however, free secretory component and lactoferrin inhibited the bacterial adhesion and interacted with several bacterial proteins. Our data indicated that glycoproteins such as free secretory component and lactoferrin could act as protective compounds against infant enteric diseases, possibly binding to bacterial surface and blocking adhesion, the primordial step of S. typhimurium infection. PMID- 17116984 TI - High efficiency cross-reactive monoclonal antibody production by oral immunization with recombinant norwalk virus-like particles. AB - Monoclonal antibodies (MAbs) are important for in-depth antigenic characterization and diagnosis of infections with human caliciviruses that cause almost all outbreaks of nonbacterial gastroenteritis. We compared different routes of immunization with nonreplicating virus-like particles (VLPs) from recombinant Norwalk virus (rNV) and recombinant Mexico virus (rMX) administered to BALB/c mice to determine the efficiency of hybridoma production. Oral immunization with VLPs without adjuvant resulted in high yields of MAb-secreting hybridomas (90%) to these VLPs of IgG (61%), IgM (29%) and IgA (10%) isotypes. Fusions with mesenteric lymph node lymphocytes yielded MAbs of various subclasses including IgG2a, IgG3, IgM and IgA. These results suggest that an immunization route that mimics the natural route of viral infection pathway may facilitate MAb technology by increasing the yields of antibody secreting hybridoma cells. PMID- 17116985 TI - Mycobacterium kumamotonense Sp. Nov. recovered from clinical specimen and the first isolation report of Mycobacterium arupense in Japan: Novel slowly growing, nonchromogenic clinical isolates related to Mycobacterium terrae complex. AB - Three mycobacterium strains isolated from clinical specimens in Japan were provisionally assigned to the genus Mycobacterium based on their phenotypical characteristics. These isolates were further investigated to determine their specific taxonomic statuses. Mycolic acid analysis and 16S rRNA gene, rpoB, and hsp65 sequence data for the isolates showed that they are most similar to M. terrae complex. DNA-DNA hybridization studies indicated that the three strains were of two species and were distinguishable from M. terrae, M. nonchromogenicum, and M. hiberniae. Therefore, these strains represent two novel species within the genus Mycobacterium. However, one potential new species should have been considered as M. arupense with the 16S rRNA gene and hsp65 sequences similarities of 99.8% and 100% respectively; it was isolated from human specimens in the United States and was proposed in June 2006 as a new species. This report describes the first isolation of M. arupense in Japan, suggesting that the organism is clinically relevant. In addition, we propose the novel species designation Mycobacterium kumamotonense sp. nov. The type strain is CST 7247(T) (=GTC 2729(T), =JCM 13453(T), =CCUG 51961(T)). PMID- 17116986 TI - Synonymy of Calyptogena solidissima with Calyptogena kawamurai (Bivalvia: Vesicomyidae) and its population structure revealed by mitochondrial DNA sequences. AB - Nucleotide sequences of part (1,101 bp) of the mitochondrial cytochrome oxidase c subunit I (COI) gene were determined for two specimens of Calyptogena kawamurai collected in Kashima Nada and Suruga Bay, respectively. These sequences were identical to each other and to those from many individuals of Calyptogena solidissima, i.e., 11 of 12 specimens from a seep area in Nankai Trough, two of 20 from hydrothermal-vent fields in Okinawa Trough, and one of 14 from a seep area on Kuroshima Knoll. The nucleotide sequences of the 5' part (about 700 bp) of the first internal transcribed spacer (ITS-1) also showed a close relationship between C. kawamurai and C. solidissima. The radiating threads on the shell surface that were emphasized in describing C. solidissima are not consistent throughout these local populations. Variation in cardinal dentition was confirmed to be intraspecific by observations of a series of specimens. The shell length height and shell length-width relationships of both species all fit a single regression line. These results suggest that C. solidissima is a junior synonym of C. kawamurai. The populations of Nankai Trough, Okinawa Trough, and Kuroshima Knoll were shown to be diverging genetically from each other. Populations of Okinawa Trough and Kuroshima Knoll are suggested to have derived independently from the most common haplotype of Nankai Trough. PMID- 17116987 TI - A taxonomic study on the leafhopper genus Scaphoidella vilbaste (Hemiptera: Cicadellidae: Deltocephalinae) from China. AB - All six species of the leafhopper genus Scaphoidella Vilbaste from China are reviewed. Among them, two new species, S. undosa sp. nov. and S. acaudata sp. nov., are described; two new combinations, Scaphoide. unihamata (Li et Kuoh) comb. nov. and Scaphoidella wideaedeaga (Wang et Li) comb. nov. are proposed; and two new synonyms are revealed, Scaphoideus inermis Cai et He (2001) syn. nov. as a junior synonym of Scaphoidella unihamata (Li et Kuoh 1993) and Scaphoideus multipunctus Li et Dai (2004) syn. nov. as a junior synonym of Scaphoidella stenopaea Anufriev (1977). A key is given to separate all six Chinese species, and illustrations of genitalia are provided. PMID- 17116988 TI - Macrostomida (Platyhelminthes: Rhabditophora) from Argentina, with descriptions of two new species of Macrostomum and of stylet ultrastructure. AB - In this work, two new species of the genus Macrostomum (M. velastylum n. sp. and M. puntapiedrensis n. sp.) are described from the littoral benthos of the Rio de la Plata estuary, Argentina. This is the first description of members of the Macrostomidae from Argentina, and of a species documented from both limnic and brackish-water environments. Macrostomum velastylum differs from its congeners in the fine details of its stylet, which are described using a new technique for stylet isolation and observation (scanning electron microscope). This method provides new information on stylet structure relative to traditional whole-mount techniques. Macrostomum puntapiedrensis differs from others species of the M. orthostylum group in the length and morphology of the distal tip of the stylet. Further details are provided on additional macrostomids from Argentina, including potential conspecifics of M. vejdovskyi, M. viride, and M. lineare. PMID- 17116989 TI - Effects of previous experience on the agonistic behaviour of male crickets, Gryllus bimaculatus. AB - Male solitary animals frequently enter aggressive interactions with conspecific individuals to protect their territory or to gain access to females. After an agonistic encounter, the loser (subordinate individual) changes its behaviour from aggression to avoidance. We investigated agonistic interactions between pairs of male crickets to understand how dominance is established and maintained. Two naive males readily entered into agonistic interactions. Fights escalated in a stereotyped manner and were concluded with the establishment of dominance. If individuals were isolated after the first encounter and placed together 15 minutes later, subordinate crickets tended to avoid any further contact with the former dominant opponent. Moreover, subordinate males also avoided unfamiliar dominant and naive opponents. They displayed aggressive behaviour only towards unfamiliar subordinate opponents. This suggests that the subordinate male change their behaviour depending on the dominance status of the opponent. Dominant crickets, in contrast, displayed aggressive behaviour towards familiar as well as unfamiliar opponents. If the interval between the first and second encounter was longer than 30 minutes, the former subordinate male showed aggressive behaviour again. However, if the subordinate cricket was paired with the same opponent three consecutive times within 45 minutes, it avoided the former dominant opponent for up to 6 hours following the third encounter. Our results suggest that the maintenance of dominance in male crickets depends largely on the behavioural change of subordinate individuals. Possible mechanisms to maintain dominance are discussed. PMID- 17116990 TI - Early development of zooxanthella-containing eggs of the corals Porites cylindrica and Montipora digitata: The endodermal localization of zooxanthellae. AB - We studied the early development of zooxanthellae-containing eggs of the scleractinian corals Porites cylindrica and Montipora digitata to elucidate how zooxanthellae become localized to the endoderm of planulae during the course of development. In both species, zooxanthellae were distributed evenly in the oocytes and delivered almost equally to the blastomeres during cleavage. In P. cylindrica, gastrulation occurred via delamination or ingression, and blastomeres containing zooxanthellae dropped into the blastocoel during gastrulation. Thus, zooxanthellae were restricted to the endodermal cells at the gastrula or early planula stage in P. cylindrica. In M. digitata, gastrulation occurred by a combination of invagination and epiboly to form a somewhat concave gastrula. Zooxanthellae were present in both endodermal and ectodermal cells of early planulae, but they disappeared from the ectoderm as the planulae matured. In our previous study on two species of Pocillopora, we found that zooxanthellae were localized in eggs as well as in embryos, and that blastomeres containing zooxanthellae later dropped into the blastocoel to become restricted to the endoderm (Hirose et al., 2000). The timing and mechanism of zooxanthella localization and types of gastrulation differed among species belonging to the three genera. These results suggest that zooxanthella localization in the embryos reflects the timing of the determination of presumptive endoderm cells and/or specificity of zooxanthellae toward presumptive endoderm cells. PMID- 17116991 TI - Composition, diversity, and spatial relationships of anurans following wetland restoration in a managed tropical forest. AB - We investigated the composition, diversity, and patterns of spatial use of an anuran community following a wetland restoration project in a managed tropical monsoon forest in southern Taiwan. Anurans comprising eight species in seven genera and four families reached a mean density of 0.025+/-0.004 anurans m(-2) within a year. The three most important species in terms of frequency of occurrence and relative abundance all had an early appearance; only the most abundant Fejervarya limnocharis (62.2% of total accounts), however, remained present throughout the entire year. The distribution of anurans observed among habitat zones was non-random, with most records occurring in densely planted (42.8%) and running-water (23.9%) zones, and the fewest in a buffer zone adjacent to a paved road. Mean numbers of anurans were correlated with the mean percent vegetation cover among individually divided small pools. Microhyla ornate, M. heymonsi, Polypedates megacephalus, and F. limnocharis appeared to be more heterogeneous in their use of space than Buergeria japonicus. Species differed in their most frequently used habitat zones within the wetland site, with the pairs F. limnocharis and P. megacephalus, and M. ornate and Bufo melanostictus, exhibiting similar respective distributions among zones. Our study demonstrates the value of even a small, isolated wetland in contributing to and maintaining regional amphibian diversity. Patterns of spatial relationships of this anuran community have important implications for the conservation of local populations across species. PMID- 17116992 TI - Phylogenetic analysis of hampala fishes (subfamily Cyprininae) in Malaysia inferred from partial mitochondrial cytochrome B DNA sequences. AB - This study examined 396 base pairs of the mitochondrial cytochrome b gene from 110 individuals belonging to the genus Hampala, a group of freshwater cyprinids that inhabit Southeast Asia. The samples were taken from various locations throughout Sarawak, Sabah, and peninsular Malaysia. The nucleotide sequences were subjected to phylogenetic analyses by using the neighbor-joining, maximum parsimony, and maximum likelihood methods. All three methods revealed the reciprocally monophyletic relationship of Hampala macrolepidota to the other Hampala forms, thus strongly supporting its status as a distinct species. Phylogenetic analysis also discovered the existence of two H. bimaculata lineages endemic to Borneo: (1) a newly identified species from the southern and central part of Sarawak assigned as H. bimaculata Type A and (2) the previously described H. bimaculata from northern Sarawak and the west coast of Sabah assigned as H. bimaculata Type B. However, the status of H. sabana and an intermediate form were not elucidated. The results suggest that the intermediate form from the Tawau population is actually a subpopulation of H. sabana, while the highly divergent intermediate form from Kalabakan could represent a cryptic species. The sharing of H. macrolepidota haplotypes in the southern peninsular Malaysia and southern and central Sarawak samples (Hm1 and Hm2) reflected the recent disconnection of the two regions, during the late Pleistocene. Overall, the partial sequence of the mitochondrial cytochrome b gene was useful for resolving the phylogenetic relationships among Hampala fishes in Malaysia. PMID- 17116993 TI - Adaptive changes in TEF-1 gene expression during cold acclimation in the medaka. AB - How animals adaptively respond to a cold or hot environment has been questioned for a long time. Recently, with the aid of microarray analysis, various temperature-sensitive genes have been identified in several species. However, a definitive hypothesis regarding the mechanism of adaptation has not been proposed. In the present study, we surveyed, in medaka (Oryzias latipes), genes for which the level of expression changes depending on the surrounding temperature. A messenger RNA differential display of medaka muscle total RNA revealed one such gene encoding transcription enhancer factor-1 (TEF-1). In medaka muscle, the TEF-1 gene produces two splicing variants, TEF-1A and TEF-1B mRNAs. During cold acclimation, the mRNA level of TEF-1A decreased, whereas that of TEF-1B increased. We also found that three putative downstream genes of TEF-1, two for myosin heavy chain (MyHC) and one for troponin T (TnT), a specific group of muscle proteins, were transcribed in a temperature-dependent manner. These results suggest that the transcription of MyHC and/or TnT is regulated by TEF-1 and that these molecules participate in muscle reconstruction during temperature adaptation in fish. PMID- 17116994 TI - Localization of vanabins, vanadium-binding proteins, in the blood cells of the vanadium-rich ascidian, Ascidia sydneiensis samea. AB - Some species of the family Ascidiidae accumulate vanadium in concentrations in excess of 350 mM, which is about 10 (7)-fold higher than the concentration of vanadium in seawater. In these species, signet ring cells with a single large vacuole in which vanadium ions are contained function as vanadium-accumulating cells. These have been termed vanadocytes. We recently isolated five vanadium binding proteins, which we named Vanabin1, Vanabin2, Vanabin3, Vanabin4, and VanabinP, from vanadocytes of the vanadium-rich ascidian Ascidia sydneiensis samea. In this study, we analyzed localization of the Vanabins in the blood cells of A. sydneiensis samea using monoclonal antibodies and confocal microscopy. The Vanabin1 and Vanabin2 proteins were found in the cytoplasm and/or in some organelles of vanadocytes. Vanabin3 was also detected in the cytoplasm, while Vanabin4 was found exclusively in the cytoplasmic membrane. PMID- 17116995 TI - A taxonomic study of Oconnorella (Enchytraeidae, Oligochaeta) from Changbaishan Mountain, China. AB - We describe three enchytraeid species, including two new species, from Mt. Changbaishan, Jilin Province, northeastern China. Oconnorella cheni sp. nov. is characterized by a simple spermatheca; absence of the oesophageal appendages and seminal vesicle; and nephridia from 6/7, with a distinct funnel and the efferent duct arising from the anterior part of the postseptale. Oconnorella globula sp. nov. is distinguished by a spermatheca with two diverticula; lack of oesophageal appendages and seminal vesicle; and nephridia from 6/7, with distinct funnel and the efferent duct arising from the mid-ventral or posteroventral part of the postseptale. We redescribe Oconnorella changbaishanensis (Xie et al., 2000) from type and live specimens, and amend some characters that cannot be investigated clearly from mounted specimens. We revise the generic diagnosis of Oconnorella. PMID- 17116996 TI - Taxonomic problems in the Drosophila melanica species group (Diptera: Drosophilidae) from Southern China, with special reference to karyotypes and reproductive isolation. AB - Karyotypes and reproductive isolation were studied in two allopatric populations of Drosophila tsigana, one from Guizhou Province in southern China and the other from Hokkaido in northern Japan, and in one population of a closely related species, D. longiserrata, from Guizhou. In metaphase plates of larval brain cells, both geographic strains of Drosophila tsigana showed 2n=10 chromosomes, with 2 pairs of metacentric (V-shape), 2 pairs of acrocentric (R-shape), and 1 pair of dot-liked (D-shape) chromosomes. Drosophila longiserrata showed the same number, 10 chromosomes, comprising 2V, 1J (sub-metacentric chromosome), 1R, and 1D. X chromosomes of both species were acrocentric, the presumed ancestral form. Premating isolation was complete between D. tsigana and D. longiserrata, and successful mating was also limited in crosses between the two geographic populations of D. tsigana, especially in crosses between Japanese (JP) females and Guizhou (GZ) males. F1 hybrids were obtained only from crosses between GZ females and JP males, and fertilities of both F1 females and males were quite incomplete. The results of morphological observations, karyotypic analyses, and crossing experiments clearly showed that the GZ and JP populations of "D. tsigana" were highly divergent from each other and that each population should be recognized as a biologically valid species. The present morphological observations and chromosomal analyses, together with the original descriptions, strongly suggest that "Guizhou D. tsigana" might be conspecific with D. bisetata Toda, 1988 from Myanmar, and that D. longiserrata might be conspecific with D. afer Tan, Hsu, and Sheng, 1949 from Meitan, Guizhou. PMID- 17116997 TI - A new mite of the winterschmidtiid genus Ensliniella (Acari: Astigmata) associated with the vespid wasp Allodynerus mandschuricus (Insecta: Hymenoptera) from Japan. AB - A new species of Ensliniella is described from deutonymphs from two localities in central Japan as the seventh species of the genus. Ensliniella asiatica sp. nov., found in the nests and acarinaria (mite chambers) of the vespid wasp Allodynerus mandschuricus, differs from the most similar species, E. kostylevi, in having a more reduced, shorter, subconical solenidion psi on tibia IV and ensiform setae e and r on tarsus III. It is readily distinguished from the remaining five known congeners in having a solenidion omega2 on tarsus I. This is the second species of Ensliniella known from Japan. PMID- 17116998 TI - Research and the survival of physical medicine and rehabilitation. PMID- 17116999 TI - Superimposed electrical stimulation: assessment of voluntary activation and perceived discomfort in healthy, moderately active older and younger women and men. AB - OBJECTIVE: An inability of the nervous system to fully activate the muscle is one factor that can contribute to age-related muscle weakness. Superimposed electrical stimulation can be used to determine voluntary muscle activation (VA). The aim of this study was to assess VA of the quadriceps muscle in healthy older and younger subjects. DESIGN: Electrical stimulation causes moderate discomfort in younger subjects, but no study has assessed discomfort in older subjects. The quadriceps muscle in 20 moderately active older subjects (mean age, 75 yrs) and 12 younger subjects (mean age, 25 yrs) was stimulated during two maximal voluntary contractions using a 100-Hz pulse train. A visual analog scale for pain (VAS-pain) was used to evaluate discomfort. RESULTS: Ability to activate the quadriceps muscle was generally very high, and there was no significant difference between the older (mean, 0.96) and younger (mean, 0.98) subjects. Discomfort did not differ between the older (mean VAS-pain score, 41 mm) and younger (mean VAS-pain score, 37 mm) subjects. CONCLUSIONS: Our results indicate that healthy, moderately active older subjects have the ability to almost complete VA of the quadriceps muscle and that discomfort during electrical stimulation is generally moderate. PMID- 17117000 TI - The effect of abdominal massage on bowel function in patients with spinal cord injury. AB - OBJECTIVES: To investigate the effect of abdominal massage on clinical aspects of bowel dysfunction and colonic transit time in patients with spinal cord injury. Twenty-four patients were placed on a standard bowel program (phase I), after which abdominal massage was added to the regimen (phase II). Parameters of gastrointestinal system function and colonic transit times were evaluated. DESIGN: Uncontrolled clinical study. RESULTS: Eleven (45.8%) of the 24 patients had abdominal distention, and 10 (41.7%) had fecal incontinence in phase I; corresponding results for phase II were three (12.5%) and four (16.7%) (P = 0.008 and 0.031, respectively). There were no significant differences between the proportions of patients with difficult intestinal evacuation or abdominal pain or in mean time required for bowel evacuation in phase I vs. phase II. The mean frequencies of defecation in phases I and II were 3.79 +/- 2.15 (2.75-4.55) and 4.61 +/- 2.17 (3.67-5.54) bowel movements per week, respectively (P = 0.006). Mean total colonic transit time decreased from 90.60 +/- 32.67 (75.87-110.47) hrs in phase I to 72 +/- 34.10 (58.49-94.40) hrs in phase II (P = 0.035). CONCLUSIONS: Abdominal massage has positive effects on some clinical aspects of neurogenic bowel dysfunction in patients with spinal cord injury. PMID- 17117001 TI - The effects of a whole-body vibration program on muscle performance and flexibility in female athletes. AB - OBJECTIVE: This randomized controlled study was designed to investigate the short term effects of an 8-wk whole-body vibration protocol on muscle performance and flexibility in female competitive athletes. DESIGN: Twenty-six young volunteer female athletes (ages 21-27 yrs) were randomized to either the vibration group or control group. The vibration intervention consisted of an 8-wk whole-body vibration 3 times a week employed by standing on a vertical vibration platform. As outcome measures, three performance tests (counter-movement jump, extension strength of lower extremities with an isokinetic horizontal leg press, and a sit and-reach test for flexibility) were performed initially and after 8 wks. RESULTS: A total of 24 athletes completed the study properly. In the vibration group (n = 13) whole-body vibration induced significant improvement of bilateral knee extensor strength (P < 0.001), counter-movements jump (P < 0.001), and flexibility (P < 0.001) after 8 wks of training. No significant changes were found for all the outcome measures for the control group (n = 11). CONCLUSIONS: Whole-body vibration is a suitable training method to improve knee extension maximal strength, counter-movement jump, and flexibility in a young female athlete if it is properly designed. Not only do the optimal frequency, amplitude, and g-forces need to be identified but also the level of muscle activation that would benefit more from vibration stimulation. The improvement of flexibility is important not only for performance but also for the prevention of muscle-tendon injury. PMID- 17117002 TI - Hybrid power-assisted functional electrical stimulation to improve hemiparetic upper-extremity function. AB - OBJECTIVE: The effect on spastic hemiparesis of a hybrid therapy consisting of functional electrical stimulation and block therapy was examined. DESIGN: Sixteen consecutively enrolled stroke patients who had spastic upper-extremity impairments more than 1 yr after stroke were recruited for this nonblinded randomized controlled trial. Patients underwent hybrid functional electrical stimulation therapy on their extensor carpi radialis longus and brevis, extensor digitorum communis, and extensor indicis proprius muscles once or twice a week for 4 mos after motor point blocks at the spastic finger flexor muscles. Surface electrodes picked up the electromyography signal and stimulated those muscles in proportion to the integrated electromyography signal obtained by the functional electrical stimulation device. The root mean square of the extensor carpi radialis longus and extensor digitorum communis maximum voluntary electromyography, active range of motion of wrist extension, finger extension, Modified Ashworth Scale, and two clinical tests were examined before and after training. RESULTS: Root mean square, active range of motion, Modified Ashworth Scale, and two clinical tests showed marked improvement in all patients as compared with the control subjects. CONCLUSIONS: The hybrid therapy was effective for patients with chronic spastic hemiparesis. Proprioceptional sensory feedback may have an important role in power-assisted functional electrical stimulation therapy. PMID- 17117003 TI - Effect of physical fitness on prosthetic ambulation in elderly amputees. AB - OBJECTIVE: To evaluate the physical fitness and prosthetic ambulation ability of high-level (hip disarticulation, transfemoral) lower-limb amputees aged 60 and above, and to investigate the level of fitness required for successful prosthetic ambulation. DESIGN: The test subjects were 49 amputees aged 60 and above. Subjects were permitted to use any necessary ambulatory aids and were asked to walk at their most comfortable walking speed for evaluation of prosthetic ambulation ability. An incremental exercise test was performed to evaluate physical fitness. RESULTS: The average %VO(2max) value for the successful user group was 64.4%, and the average %VO(2max) value for the failed user group was 44.8%, a statistically significant difference (P < 0.01). Of the amputees with %VO(2max) > or =50, 93.7% were in the successful user group. Of the amputees with %VO(2max) <50, only 23.5% were in the successful user group. CONCLUSION: This study showed that the ability to sustain an exercise intensity of > or =50%VO(2max) can be regarded as a guideline value for the level of fitness required for successful prosthetic ambulation in elderly amputees. PMID- 17117004 TI - Sacroiliac joint pain: anatomy, biomechanics, diagnosis, and treatment. AB - The sacroiliac joint is an underappreciated cause of low back and buttock pain. It is thought to cause at least 15% of low back pain. It is more common in the presence of trauma, pregnancy, or in certain athletes. The pelvic anatomy is complex, with the joint space being variable and irregular. The joint transmits vertical forces from the spine to the lower extremities and has a role in lumbopelvic dynamic motion. History and physical examination findings can be helpful in screening for sacroiliac joint pain, but individual provocative maneuvers have unproven validity. Fluoroscopically guided injections into the joint have been found to be helpful for diagnostic and therapeutic purposes. Conservative treatment, which also can include joint mobilization, antiinflammatory medicines, and sacroiliac joint belts, generally is effective. Surgical arthrodesis should be considered a procedure of last resort. PMID- 17117005 TI - Airbag-mediated pediatric atlanto-occipital dislocation. AB - Pediatric atlanto-occipital dislocation (AOD) most commonly occurs in victims of high-velocity motor vehicle collisions and usually results in death. The mechanical and anatomic predisposing factors for AOD in children have been well documented. With the introduction of passenger-side air bags in automobiles, reports have documented an increased incidence of pediatric cervical spine injuries, including AOD, in low-speed motor vehicle collisions. Although long term survival in AOD is so unusual that there is no separate literature regarding its physiatric treatment, it is generally treated like any other high cervical spinal cord injury. Recent advances in adaptive technology have aided greatly in management. Despite benefits that adaptive technology affords, the best treatment for AOD is still prevention. For this reason, children under the age of 13 should be restrained backseat passengers in automobiles. PMID- 17117007 TI - SCI sleep disordered breathing vs. hypoventilation? PMID- 17117008 TI - Re: Electrical twitch-obtaining intramuscular stimulation in lower back pain: a pilot study. PMID- 17117010 TI - Acute median-nerve compression caused by calcifying aponeurotic fibroma. PMID- 17117011 TI - Candidate HIV-1 Tat vaccine development: from basic science to clinical trials. PMID- 17117012 TI - Identification of novel consensus CD4 T-cell epitopes from clade B HIV-1 whole genome that are frequently recognized by HIV-1 infected patients. AB - OBJECTIVE: To identify promiscuous and potentially protective human CD4 T-cell epitopes in most conserved regions within the protein-coding genome of HIV-1 clade B consensus sequence. DESIGN: We used the TEPITOPE algorithm to screen the most conserved regions of the whole genome of the HIV-1 subtype B consensus sequence to identify promiscuous human CD4 T-cell epitopes in HIV-1. The actual promiscuity of HLA binding of the 18 selected peptides was assessed by binding assays to nine prevalent HLA-DR molecules. Synthetic peptides were tested with interferon-gamma ELISPOT assays on peripheral blood mononuclear cells (PBMC) from 38 HIV-1 infected patients and eight uninfected controls. RESULTS: Most peptides bound to multiple HLA-DR molecules. PBMC from 91% of chronically HIV-1 infected patients recognized at least one of the promiscuous peptides, while none of the healthy controls recognized peptides. All 18 peptides were recognized, and each peptide was recognized by at least 18% of patients; 44% of the patients recognized five or more peptides. This response was not associated to particular HLA-DR alleles. Similar responses were obtained in CD8 T-cell-depleted PBMC. CONCLUSION: In silico prediction of promiscuous epitopes led to the identification of naturally immunodominant CD4 T-cell epitopes recognized by PBMC from a significant proportion of a genetically heterogeneous patient population exposed to HIV-1. This combination of CD4 T-cell epitopes - 11 of them not described before - may have the potential for inclusion in a vaccine against HIV 1, allowing the immunization of genetically distinct populations. PMID- 17117013 TI - Increased carotid intima-media thickness in HIV patients is associated with increased cytomegalovirus-specific T-cell responses. AB - OBJECTIVES: HIV-infected subjects are at increased risk for myocardial infarction. The mechanism of this increased risk remains unclear. Since cytomegalovirus (CMV) infection has been associated with accelerated atherosclerosis in the transplant population and immune responses against CMV may be altered by HIV disease, we hypothesized that enhanced T-cell responses against CMV would be associated with increased atherosclerosis in subjects with HIV. METHODS: We measured high-sensitivity C-reactive protein (hs-CRP), T-cell activation, CMV-specific T-cell responses, and carotid artery intima-media thickness (IMT) in 93 HIV-infected subjects and in 37 uninfected controls. RESULTS: The mean age of the HIV-infected subjects was 48 years and 85 (91%) were male. The median carotid IMT was higher in the HIV-infected group compared to the uninfected group (0.95 mm versus 0.68 mm, P < 0.001). This difference remained significant after controlling for all traditional risk factors. Compared to HIV negative controls, HIV-infected subjects had higher median levels of hs-CRP (P = 0.05), higher levels of CD4 and CD8 T-cell activation (P < 0.0001) and higher CMV specific interferon-gamma CD8 T-cell responses (P < 0.0001). CMV-specific T-cell responses, but not hs-CRP and T-cell activation, were independently associated with higher carotid IMT (P = 0.001). CONCLUSIONS: HIV-infected subjects had thicker carotid IMT compared to controls. While HIV patients also had higher T cell activation, hs-CRP levels, and CMV-specific T-cell responses, only CMV specific T-cell responses were independently associated with IMT. Accelerated atherosclerosis in HIV patients may be mediated by heightened CMV-induced immune responses. PMID- 17117014 TI - Genomic imbalances in AIDS-related lymphomas: relation with tumoral Epstein-Barr virus status. AB - BACKGROUND: The pathologic heterogeneity of AIDS related lymphomas (ARL) reflects several pathogenic mechanisms: chronic antigenic stimulation, Epstein-Barr virus (EBV) infection, and genomic abnormalities. Genetic abnormalities, known to play a major role in lymphomas of non-immunocompromised patients, are not well characterized in ARL. OBJECTIVE: Characterization of the DNA copy number change (CNC) in ARL and comparison of our findings with tumoral EBV and immune status. DESIGN AND METHODS: We have studied by comparative genomic hybridization (CGH), 28 ARL well characterized for histopathologic, clonality and EBV findings. RESULTS: DNA-CNC were detected in 50% of cases. Gains of chromosomal material were much more frequent than losses and involved chromosomes 9p, 11q, 12q, 17q, and 19q recurrently. DNA-CNC tended to be more frequent in EBV-positive lymphomas with latency type II/III than in EBV-positive latency I or EBV-negative lymphomas. Most chromosomal regions affected in HIV-related lymphoma were similar to those already reported in HIV-negative lymphomas. CONCLUSION: This CGH study allowed the identification of non-random chromosomal alterations in ARL. The results suggested an inverse relationship between EBV infection (latency II/III), associated with deep acquired immune suppression, and the number of chromosomal alterations which may be explained by a direct role of viral proteins in lymphomagenesis by activation of signalling pathways without needing several genomic alterations. PMID- 17117015 TI - Sequential priming and boosting with heterologous HIV immunogens predominantly stimulated T cell immunity against conserved epitopes. AB - BACKGROUND: The effort to develop an effective preventive vaccine against HIV-1 infection is challenged by the wide genetic diversity of HIV-1 among different isolates. OBJECTIVES: To explore a new vaccination strategy by using heterologous HIV immunogens derived from different clades for sequential priming and boosting. METHODS: HIV Env and Gag immunogens derived from Thailand B (B'), C/B' recombinant and A/E recombinant were selected as these three clades account for 29%, 45% and 15% of HIV-1 prevalence in China, respectively. Three humanized fusion genes of env and gag derived from those three clades were synthesized and inserted into DNA and recombinant Tiantan vaccinia vectors as model vaccines. C57BL/6 and Balb/c mice were used as animal model. Peptides spanning the entire Env and Gag were used as stimuli and Elispot assay was used to assess the T cell immunity. RESULTS: Sequential priming and boosting was observed with heterologous HIV immunogens predominantly stimulated T cell immunity against conserved epitopes, whereas a single vaccine derived from one clade or the mixture of multiple vaccines from different clades primarily raised T cells against less conservative or non-conservative epitopes. CONCLUSIONS: This is the first demonstration of a practical strategy to raise immune responses against conserved epitopes. This strategy has important implications for vaccine development against HIV and other pathogens that have high genetic diversity, such as influenza. PMID- 17117016 TI - Impact of suppressive herpes therapy on genital HIV-1 RNA among women taking antiretroviral therapy: a randomized controlled trial. AB - OBJECTIVE: To demonstrate a causal relationship between herpes simplex virus 2 (HSV-2) and increased genital HIV-1-RNA shedding in women on HAART. DESIGN: A randomized, double-blind, placebo-controlled trial of herpes-suppressive therapy (valacyclovir 500 mg twice a day) in HIV-1/HSV-2-infected women taking HAART in Burkina Faso. METHODS: Participants were followed for a total of 12 biweekly visits before and after randomization. The presence and frequency of genital and plasma HIV-1 RNA, and of genital HSV-2 were assessed using summary measures, adjusting for baseline values. Random effect linear regression models were used to assess the impact of treatment on genital and plasma viral loads among visits with detectable virus. RESULTS: Sixty women were enrolled into the trial. Their median CD4 lymphocyte count was 228 cells/mul, and 83% had undetectable plasma HIV-1 RNA at baseline. Valacyclovir reduced the proportion of visits with detectable genital HSV-2 DNA [odds ratio (OR) 0.37, 95% confidence interval (CI) 0.13, 1.05], but had no significant impact on the frequency (OR 0.90, 95% CI 0.31, 2.62) or quantity (reduction of 0.33 log copies/ml, 95% CI -0.81, 0.16) of genital HIV-1 RNA. However, according to pre-defined secondary analyses restricted to women who shed HIV-1 at least once in the baseline phase, valacyclovir reduced both the proportion of visits with detectable HIV-1 shedding (OR 0.27, 95% CI 0.07, 0.99) and the quantity of genital HIV-1 RNA during these visits (-0.71 log10 copies/ml, 95% CI -1.27, -0.14). CONCLUSION: HSV-2 facilitates residual genital HIV-1 replication among dually infected women taking HAART despite HIV-1 suppression at the systemic level. PMID- 17117017 TI - Long-term survival and immuno-virological response of African HIV-1-infected children to highly active antiretroviral therapy regimens. AB - BACKGROUND: In Africa, facing the scaling-up of HAART, there is an urgent need to monitor accurately the long-term benefits of these lifelong treatments. METHODS: Survival and immuno-virological response were assessed for 78 children in the ANRS 1244/1278 Children's cohort (Abidjan, Cote d'Ivoire) who were enrolled from October 2000 for treatment with HAART and followed to September 2004. Initial HAART consisted of two nucleoside reverse transcriptase inhibitors with either nelfinavir (NFV) or efavirenz (EFV). For the comparison of immunological and virological responses, CD4 cell counts and HIV-1 RNA viral load were assessed by performing time-point specific and longitudinal data analysis. RESULTS: At baseline, the median CD4 cell percentage was 7.5% and the median HIV-1 RNA viral load was 5.37 log10 copies/ml. The survival probability was high (0.86 at month 42; 95% confidence interval, 0.77-0.92) with no difference according to whether the HAART regimen contained NFV or EFV. At 36 and 42 months of follow-up, an immune recovery was observed with median CD4 cell percentages reaching 23.1% and 24.8%, respectively, with no difference according to the HAART regimen (longitudinal data analysis). At the same time points, a sustained viral suppression was also obtained, with undetectable viral load achieving in 46.5% and 45.0%, respectively, regardless of whether the HAART regimen. CONCLUSION: This study demonstrates the durability of both clinical and biological response to HAART in African children. PMID- 17117018 TI - Varicella vaccination in HIV-1-infected children after immune reconstitution. AB - BACKGROUND: HIV-1-infected children have an increased risk of severe chickenpox. However, vaccination is not recommended in severely immunocompromised children. OBJECTIVE: Can the live-attenuated varicella zoster virus (VZV) Oka strain be safely and effectively given to HIV-1-infected children despite previously low CD4 T-cell counts? METHODS: VZV vaccine was administered twice to 15 VZV seronegative HIV-1-infected children when total lymphocyte counts were greater than 700 lymphocytes/microl, and six HIV-negative VZV-seronegative siblings. Weekly clinical follow-up and sampling were performed. RESULTS: None of the children developed any clinical symptom or serious adverse reaction after immunization. Only nine (60%) of the HIV-1-infected children had VZV-specific antibodies after two immunizations, whereas 100% of the siblings seroconverted. Age at baseline was negatively correlated with the VZV IgG titre at 6 weeks after the second vaccination in HIV-1-infected children. VZV-specific antibody titres after two immunizations were at a similar level to those found after wild-type infection in non-vaccinated HIV-1-infected patients, but significantly lower than in HIV-negative siblings. Importantly, VZV-specific T-cell responses increased after vaccination and were comparable in both groups over time. Documented wild type VZV contact in three vaccinated patients did not result in breakthrough infections. CONCLUSION: VZV vaccination of previously immunocompromised HIV-1 infected children was safe. Vaccination induced specific immune responses in some of the vaccinated HIV-1-infected children, suggesting that previously immunocompromised individuals are protected against severe forms of varicella. PMID- 17117019 TI - The level of persistent HIV viremia does not increase after successful simplification of maintenance therapy to lopinavir/ritonavir alone. AB - OBJECTIVE: To determine whether the level of persistent HIV-1 viremia is affected by simplifying standard antiretroviral therapy to lopinavir/ritonavir (LPV/r) alone. DESIGN: Measurement of HIV-1 RNA levels < 50 copies/ml in longitudinal plasma samples from 41 of 42 subjects enrolled in the 'Only Kaletra' study that compared maintenance therapy with LPV/r alone to standard of care (SOC) with two nucleoside reverse transcriptase inhibitors (NRTI) and LPV/r. METHODS: Plasma samples for each subject from study screening to week 48 were tested using a modified Roche Amplicor HIV-1 RNA assay with a quantification limit of 3 copies/ml. RESULTS: Median plasma HIV-1 RNA values at baseline and weeks 4, 8, 12, 24 and 48 were not significantly different between the LPV/r alone and the SOC arms, being 5.1 versus 3.0 (P = 0.29), 4.5 versus 2.9 (P = 0.44), 3.3 versus 2.9 (P = 0.99), 1.9 versus 1.0 (P = 0.68), 3.7 versus 3.6 (P = 0.49), and 2.8 versus 1.6 copies/ml (P = 0.78), respectively. In the 17 of 21 subjects who maintained virus suppression < 50 copies/ml on LPV/r alone, median HIV-1 RNA values did not increase significantly from baseline at any time point after discontinuing NRTI, in comparison to the three subjects with virologic failure whose median HIV-1 RNA levels began to rise at week 8. CONCLUSIONS: The level of persistent viremia did not increase after stopping NRTI therapy among subjects who maintained virus suppression < 50 copies/ml on LPV/r alone through 48 weeks. This supports further studies of induction-simplification therapy for treatment of HIV-1 infection including the identification of factors predicting success or failure of simplified therapy. PMID- 17117020 TI - Human papillomavirus types among women infected with HIV: a meta-analysis. AB - BACKGROUND: HIV-positive women have a high prevalence of human papillomavirus (HPV) infection and are infected with a broader range of HPV types than HIV negative women. It is not known to what extent these different types are associated with high-grade squamous intraepithelial lesions (HSIL) and cancer. METHODS: Meta-analysis of HPV type-specific prevalence among HIV-positive women, stratified by geographical region and by cervical cytology: normal, atypical squamous cells of undetermined significance/low-grade squamous intraepithelial lesions (ASCUS/LSIL) or HSIL. RESULTS: In 20 studies, 5578 HIV-positive women were identified, largely from North America but also Africa, Asia, Europe and South/Central America. For 3230 with no cytological abnormalities, prevalence was 36.3% for any HPV and 11.9% for multiple HPV types. The six most common high-risk HPV types were 16 (4.5%), 58 (3.6%), 18 (3.1%), 52 (2.8%), 31 (2.0%) and 33 (2.0%). HPV16 was also the most common type in 2053 HIV-positive women with ASCUS/LSIL and 295 with HSIL. Those with HSIL were significantly less likely to be infected with HPV16 (odds ratio, 0.6; 95% confidence interval, 0.4-0.7) than the general female population with HSIL. In contrast, HIV-positive women with HSIL were significantly more likely to be infected with HPV types 11, 18, 33, 51, 52, 53, 58 and 61, and with multiple HPV types. CONCLUSIONS: The proportion of HIV-positive women with HPV16 rose with increasing severity of cervical lesions. Nevertheless, HPV16 remained underrepresented in HIV-positive women with HSIL, who showed a higher proportion of other HPV types and multiple types compared with the general female population with HSIL. PMID- 17117021 TI - Maternal antiretroviral drugs during pregnancy and infant low birth weight and preterm birth. AB - OBJECTIVE: To determine the relationship between maternal antiretroviral regimens during pregnancy and adverse infant outcomes [low birth weight (LBW) and preterm birth]. The a priori hypothesis was that protease inhibitor (PI)-containing regimens are associated with an increased risk of LBW and preterm birth. DESIGN: Prospective cohort study of HIV-1-infected women and their infants (NISDI Perinatal Study). METHODS: Data were analysed from 681 women receiving at least one antiretroviral drug [in order of increasing complexity: one or two nucleoside reverse transcriptase inhibitors (1-2 NRTI), two NRTI plus one non-nucleoside reverse transcriptase inhibitor (NNRTI) (HAART/NNRTI), or two NRTI plus one PI (HAART/PI)] for at least 28 days during pregnancy, and who delivered live born, singleton infants with known birth weight and gestational age by 1 March 2005. Multivariable logistic regression modeling was used to assess the relationship of maternal ART with LBW and with preterm birth, adjusting for covariates. RESULTS: The incidence of LBW and preterm birth, respectively, was 9.6% and 7.4% (1-2 NRTI), 7.4% and 5.8% (HAART/NNRTI), and 16.7% and 10.6% (HAART/PI). There was no statistically significant increased risk of LBW [adjusted odds ratio (AOR), 1.5; 95% confidence interval (95% CI), 0.7-3.2] or preterm birth (AOR, 1.1; 95% CI, 0.5-2.8) among women who received HAART/PI compared with women receiving 1-2 NRTI. CONCLUSIONS: Among a population of HIV-1-infected women in Latin America and the Caribbean, maternal receipt of PI-containing ART regimens during pregnancy was not associated with a statistically significant increase in risk of LBW or preterm birth. PMID- 17117022 TI - Risk factors for high early mortality in patients on antiretroviral treatment in a rural district of Malawi. AB - OBJECTIVES: Among adults started on antiretroviral treatment (ART) in a rural district hospital (a) to determine the cumulative proportion of deaths that occur within 3 and 6 months of starting ART, and (b) to identify risk factors that may be associated with such mortality. DESIGN AND SETTING: A cross-sectional analytical study set in Thyolo district, Malawi. METHODS: Over a 2-year period (April 2003 to April 2005) mortality within the first 3 and 6 months of starting ART was determined and risk factors were examined. RESULTS: A total of 1507 individuals (517 men and 990 women), whose median age was 35 years were included in the study. There were a total of 190 (12.6%) deaths on ART of which 116 (61%) occurred within the first 3 months (very early mortality) and 150 (79%) during the first 6 months of initiating ART. Significant risk factors associated with such mortality included WHO stage IV disease, a baseline CD4 cell count under 50 cells/mul and increasing grades of malnutrition. A linear trend in mortality was observed with increasing grades of malnutrition (chi for trend = 96.1, P /= 1 visit/year for >/= 2 years). Logistic regression was used to examine predictors of referral. RESULTS: A total of 277 (33%) of 845 were referred for HCV care. Independent predictors of referral included percentage elevated alanine aminotransferase levels [adjusted odds ratio (AOR) for 10% increase,1.16; 95% confidence interval (CI), 1.10-1.22] and CD4 cell count > 350 cells/microl (AOR, 3.20; 95% CI, 2.10-4.90), while injection drug use was a barrier to referral (AOR, 0.26; 95% CI, 0.11-0.64). Overall referral rate increased from < 1% in 1998 to 28% in 2003; however, even in 2003, 65% of those with CD4 cell count > 200 cells/microl were not referred. One hundred and eighty-five (67%) of 277 referred kept their appointment, of whom 32% failed to complete a pre-treatment evaluation. Of the remaining 125, only 69 (55%) were medically eligible for treatment, and 29 (42%) underwent HCV treatment. Ninety percent of 29 were infected with genotype 1 and 70% were African American; six (21%) achieved sustained virologic response (SVR). Only 0.7% of the full cohort achieved SVR. CONCLUSIONS: Although the potential for SVR and the recent marked increase in access to HCV care are encouraging, overall effectiveness of anti-HCV treatment in this urban, chiefly African American, HCV genotype 1 HIV clinic is extremely low. New therapies and treatment strategies are an urgent medical need. PMID- 17117024 TI - The late diagnosis and consequent short-term mortality of HIV-infected heterosexuals (England and Wales, 2000-2004). AB - OBJECTIVES: To describe the proportion of HIV-infected heterosexual individuals diagnosed late and estimate how much mortality could have been prevented by earlier diagnosis. DESIGN: A population-based study using surveillance reports from England and Wales. METHODS: Calculation of proportions diagnosed late (CD4 cell count < 200 cells/mul) and short-term mortality (death within a year of diagnosis). RESULTS: A total of 16 375 heterosexual individuals were diagnosed with HIV (2000-2004): 10 503 with CD4 cell counts available at that time; 42% (4425) were diagnosed late. Late diagnosis increased with age (P < 0.01). One fifth of women diagnosed antenatally were diagnosed late compared with 42% of other women and 49% of men; 70% of all heterosexuals diagnosed were black Africans, born and infected in Africa. Of those, at least 40% were recent arrivals to the UK, and twice as many were diagnosed late as black-African heterosexuals infected in the UK. Short-term mortality was 3.2% (491/15 523); 6.1% among those diagnosed late and 0.7% among others (P < 0.01). Short-term mortality was lower among black-African compared with white heterosexuals (3.1 versus 4.5%; P < 0.01) because of diagnosis at a younger age. Earlier diagnosis would have reduced short-term mortality by 56% (249 fewer deaths) and all mortality by 32% between 2000 and 2004. CONCLUSION: Groups at high risk of late diagnosis should be targeted for health promotion activities, opportunistic screening, and removal of any barriers to testing. HIV testing in a variety of settings would reduce missed diagnoses and costs. New patient checks in primary care may provide the earliest opportunity to diagnose HIV infection among recent arrivals to the UK. PMID- 17117025 TI - Human papillomavirus genotypes among women with HIV: implications for research and prevention. PMID- 17117026 TI - Measuring below the bar: what is it telling us? PMID- 17117027 TI - High levels of cervical HIV-1 RNA during early HIV-1 infection. AB - Few data are available on genital tract viral replication early after HIV-1 acquisition, when infectivity is high. We compared cervical HIV-1 RNA from 60 women with paired samples from within 90 days after HIV-1 acquisition and at viral setpoint (4-24 months). Cervical HIV-1 was higher in early compared with setpoint samples (mean 3.43 versus 2.85 log10 copies/swab, P < 0.001). After adjusting for HIV-1-plasma RNA, cervical HIV-1 RNA from 30 days or less after infection was increased by 0.45 log10 copies/swab (P = 0.006). PMID- 17117028 TI - Immune reconstitution inflammatory syndrome in the first year after HAART: influence on long-term clinical outcome. AB - Little is known about the effect of immune reconstitution inflammatory syndrome (IRIS) on the long-term clinical outcome. Of 52 opportunistic infections (OI) occurring within one year after the start of HAART in 387 HIV patients, 33 (63%) were classified as having IRIS. The patients with IRIS showed no significant difference in the AIDS event-free survival curve compared with the matched control group without OI and in contrast to non-IRIS OI. PMID- 17117029 TI - Yearly number of patients diagnosed with primary HIV-1 infection in France estimated by a capture-recapture approach. AB - We used a two-source capture-recapture method to estimate the number of patients diagnosed at the time of primary HIV infection in France between 1999 and 2002. The sources were the French PRIMO cohort and the French Hospital Database on HIV. The estimated number of patients was 325 per year, which represents only 5% (approximately 6000 cases) of all new cases diagnosed each year and only 8% of all new infections (approximately 4000 cases). PMID- 17117030 TI - Influence of pravastatin on carotid artery structure and function in dyslipidemic HIV-infected patients receiving antiretroviral therapy. AB - Forty-two pravastatin-treated HIV-positive patients and 42 sex, age, and smoking status-matched hypercholesterolemic HIV-positive patients not under lipid lowering treatment were compared for differences in intima-media thickness (IMT) of the common carotid artery (CCA) and aortic stiffness. Pravastatin had no influence on carotid artery structure and function, or aortic stiffness. Age and body mass index were independent determinants of IMT of the CCA. Mean arterial pressure, age, duration of HIV infection and protease inhibitor exposure determined aortic stiffness. PMID- 17117031 TI - Serious, multi-organ hypersensitivity to lopinavir alone, involving cutaneous mucous rash, and myeloid, liver, and kidney function. PMID- 17117032 TI - Incomplete information about AIDS on Chinese websites. PMID- 17117033 TI - A case for treating high hepatitis B DNA levels before starting HIV therapy. PMID- 17117036 TI - Impact of augmenting dialysis frequency and duration on cardiovascular function. AB - Conventional hemodialysis (CHD) only delivers 10% to 15% of renal function in a nonphysiological intermittent mode. Because it occurs nightly and is sustained over a longer dialysis time, the uremic clearance provided by nocturnal hemodialysis (NHD) far exceeds that of CHD. Increasing the dose and frequency of dialysis by NHD has been demonstrated, in both short- and long-term studies, to reverse several important risk factors for adverse cardiovascular events in patients with end-stage renal disease such as hypertension, left ventricular hypertrophy, systolic dysfunction, conduit artery stiffness, attenuated baroreflex regulation of heart rate, disturbed heart rate variability, sleep apnea, and endothelium-dependent vasodilation. In addition, the Toronto NHD experience has reported an emerging body of evidence demonstrating the benefits of NHD on anemia management, inflammation, and endothelial progenitor cell biology. The mechanism(s) by which nocturnal hemodialysis improves cardiovascular outcomes are under active investigation by our group. It is tempting to speculate that NHD has the potential to decrease endothelial/myocardial injury and restore simultaneously endothelial repair, thereby improving cardiovascular function in patients with end-stage renal disease. The objectives of the present document are (1) to review the mechanisms underlying dialysis-associated cardiovascular morbidity and (2) to describe the restorative potential of NHD on the cardiovascular system. PMID- 17117037 TI - Matching the mechanical circulatory support device to the child with heart failure. AB - A number of mechanical circulatory support devices are currently available for the treatment of children with heart failure refractory to medical therapy. Patient size, the acute versus chronic nature of the heart failure that is present, and the underlying anatomy and physiology define the clinical setting in any given case. Determining which device most effectively meets the specific needs of the clinical setting requires an understanding of the characteristics of each of the available devices. This report describes a graphic representation of the clinical setting in which pediatric heart failure occurs to provide a decision-making framework for the selection of the most appropriate mechanical circulatory support device. PMID- 17117040 TI - Toward human organ printing: Charleston Bioprinting Symposium. AB - The First Annual Charleston Bioprinting Symposium was organized by the Bioprinting Research Center of the Medical University of South Carolina (MUSC) and convened July 21, 2006, in Charleston, South Carolina. In broad terms, bioprinting is the application of rapid prototyping technology to the biomedical field. More specifically, it is defined as the layer by layer deposition of biologically relevant material. The 2006 Symposium included four sessions: Computer-aided design and Bioprinting, Bioprinting Technologies; Hydrogel for Bioprinting and, finally, a special session devoted to ongoing research projects at the MUSC Bioprinting Research Center. The Symposium highlight was the presentation of the multidisciplinary Charleston Bioengineered Kidney Project. This symposium demonstrated that bioprinting or robotic biofabrication is one of the most exciting and fast-emerging branches in the tissue engineering field. Robotic biofabrication will eventually lead to industrial production of living human organs suitable for clinical transplantation. The symposium demonstrated that although there are still many technological challenges, organ printing is a rapidly evolving feasible technology. PMID- 17117041 TI - Artificial organs: a new chapter in medical history. PMID- 17117042 TI - Report of the first U.S. patient successfully supported long term with the LionHeart completely implantable left ventricular assist device system. AB - We report our first successful long-term survivor in the United States with the LionHeart (Arrow International, Inc., Reading, PA) completely implantable left ventricular assist device system. The patient was initially deemed a poor candidate for cardiac transplantation and had inotrope-dependent, end-stage cardiac failure. The patient was supported for 13 months with this system. During this period of support, the patient returned to independent living and derived obvious benefits toward his daily activities with the completely implanted system. The device proved to be reliable during this period of support. Through lifestyle modification, the patient was ultimately deemed an appropriate candidate for heart transplantation and ultimately received successful transplantation. PMID- 17117043 TI - Shprintzen (velo-cardio-facial) syndrome: a rare case. AB - Shprintzen syndrome (velo-cardio-facial, VCFS) is a very rare morbid entity, seen in either familial or sporadic forms, with major clinical findings such as facial dysmorphism, cleft palate, cardiovascular (especially conotruncal-anomalies), mild/moderate mental retardation, or, more commonly, observed learning difficulty. Tendency to behavioral disorders and bipolar schizophrenic diseases may be present in these cases. Autosomal dominant inheritance has been reported. VCFS appears as a consequence of microdeletion in the 22q11 chromosomal band. Although each syndrome has different clinical reflections, genetically the defect is located on the same chromosome.A 4-year-old boy was admitted to our clinic with a syndromic face and the diagnosis of tetralogy of Fallot. The patient underwent total correction of the cardiac defect. Atypical facial appearance, cleft palate, and malformed hand-fingers were present. With the aid of pediatric and genetic consultation (fluorescence in situ hybridization test), Shprintzen syndrome was confirmed. Both early and late postoperative periods of the patient were uneventful. The patient was closely consulted by a specialist psychologist during and after the hospitalization period. These children can be integrated into social life earlier through early surgical intervention for cardiac defects and facial deformations as well as neurologic and/or neuropsychiatric evaluation. PMID- 17117044 TI - Symptomatic lipoma in the interventricular septum. AB - Cardiac lipomas, which are benign nonmyxomatous neoplasms of the heart, are rare and among those least often encountered. Because they normally cause no symptoms, diagnosis is often purely accidental. We report the case of a 24-year-old woman who presented with palpitations of recent onset and was found to have a lipoma attached to the left side of the interventricular septum (IVS). Transthoracic and transesophageal echocardiograms were performed and showed a mass in the left side of the IVS. During successful surgical excision of the mass, intraoperative histologic diagnosis showed the tumor was lipoma. Our review of the English literature revealed that our case is only the seventh of removal of lipoma in the IVS. The postoperative course was uneventful, and an echocardiogram taken 6 months after the operation showed no evidence of enlargement of the tumor tissue. PMID- 17117045 TI - Effect of direct hemoperfusion with a polymyxin B immobilized fiber column on high mobility group box-1 (HMGB-1) in severe septic shock: report of a case. AB - Because of the many difficult aspects in the treatment of septic shock and poor outcome of this condition, establishing the most appropriate therapeutic strategy is problematic. Recently, high mobility group box-1 (HMGB-1) has been shown to activate inflammatory responses and to be a late mediator in endotoxemia and sepsis. Therefore, we considered that it might be worthwhile to investigate the therapeutic potential of HMGB-1 blockade in cases of septic shock.Herein, we describe the case of a patient with septic shock with hepatic portal venous gas caused by intestinal obstruction. Hepatic portal venous gas is a rare condition associated with significant radiographic findings and a fatal outcome. Our patient, however, recovered from severe septic shock and was saved by the use of direct hemoperfusion with a polymyxin B immobilized fiber column (DHP-PMX). This treatment resulted in a decrease in the serum levels of endotoxin, interleukin-6 (IL-6), and HMGB-1. PMID- 17117046 TI - Aortico-left ventricular tunnel, an unusual congenital cardiac anomaly in adult: application of a new operative technique. AB - Aortico-left ventricular tunnel is a rare congenital cardiac anomaly. A 45-year old man was referred to our clinic with unstable angina pectoris. The patient had an aortico-left ventricular tunnel that had been operated on 15 years before and that now showed a recurrence. We performed a new surgical technique, including closure of orifices of the tunnel by resection of the aorta at the left coronary ostium, reconstruction of the aorta with patch plasty, and formation of a neo left main branch by applying a saphenous magna vein patch at the noncoronary cusp. In this technique, the possibility of aortic regurgitation caused by stretching and distortion of the aortic ring and leaflets by primary suture closure of tunnel is eliminated. The postoperative 2-D colored Doppler echocardiography and cardiac MRI showed an excellent result of the procedure. Coronary flow could be restored, and thus anginal symptoms disappeared. PMID- 17117047 TI - Pediatric patients with Kawasaki disease and a case report of Kitamura operation. AB - Kawasaki disease (KD), also called mucocutaneous lymph node syndrome, is an acute, self-limiting, small-vessel vasculitis with an unknown cause that affects children between the ages of 6 months and 5 years. It is the most common cause of acquired coronary artery disease in childhood. Acute myocardial infarction and coronary artery aneurysm are major complications. We present a cohort of patients with KD who were followed up and treated in the Heart Center, North Rhine Westphalia. Included is a review of important relevant items common to cases of KD, such as clinical data and management, including medical management of the acute condition and the diagnosis and management of coronary vasculitis and aneurysms as well as the application of coronary artery bypass grafting (CABG) in those conditions. Between January 2002 and January 2006, we evaluated the findings and characteristics of 18 pediatric patients with a history of KD and their long-term outcome. The acute illness occurred between the ages of 4 months and 14 years of age. Anomalies of the coronary arteries were found in 6 patients ranging in age from 5 months to 10 years. One patient had acute myocardial infarction; another underwent CABG after 5 years from disease onset at the age of 15 years. Kitamura operation was performed successfully. The other patients are still under observation.Coronary artery aneurysms and stenosis requiring surgery are rare in KD; nevertheless, CABG is the standard therapy when myocardial ischemia is detected. Kitamura operation provides good growth potential and long term graft patency. PMID- 17117048 TI - Can the therapeutic advantages of allogenic umbilical cord blood-derived stem cells and autologous bone marrow-derived mesenchymal stem cells be combined and synergized? PMID- 17117049 TI - Pulsatile flow past a cylinder: an experimental model of flow in an artificial lung. AB - The focus of this study is an experimental apparatus that serves as a model for studying blood flow in a total artificial lung (TAL), a prototype device intended to serve as a bridge to lung transplantation or that supports pulmonary function during the treatment of severe respiratory failure. The TAL consists of hollow cylindrical fibers that oxygen-rich air flows through and oxygen-poor blood flows around. Because gas diffusivity in the TAL is very small, a convection mechanism dominates the gas transport, which is why we focus on the velocity around the fibers (modeled as a 0.05-cm-in-diameter and 5-cm-long cylinder). We designed a low-speed water tunnel to study the flow mechanism around the cylinder, across which the flow is generated by a linear actuator that allows different flow patterns to mimic the flow in a TAL. We tested the flow in the test section by numerical simulation and by the particle image velocimetry method to study the flow profile. The results show a uniform flow near the centerline of the water tunnel where the cylinder is placed. This decreases the effects of free-stream turbulence in the shear layers and reduces the uncertainty in determining the flow patterns around the cylinder. Knowledge gained from the flow around one cylinder (fiber) is beneficial for understanding vortex formation around multiple cylinders. We present a summary of vortex formation behind a cylinder for Reynolds numbers (Re) of 1, 3, and 5 and Stokes numbers (Ns) of 0.18 to 0.37; results show that higher Re and Ns favor vortex formation. These findings regarding the parameter range for vortex formation may provide principles for designing artificial lungs to enhance convective mixing. We anticipate that the pulsatile flow circuit presented here can be used to mimic the flow not only in TALs but in other physiological systems. PMID- 17117050 TI - Numerical investigation on hydrodynamics and biocompatibility of a magnetically suspended axial blood pump. AB - A newly designed magnetically suspended axial blood pump is presented, in which a 5 degrees-of-freedom rotor is suspended by using two conical active magnetic bearings, each with a four-pole stator. The preferred configuration could provide a rather large moment of inertia to increase the rotating stability of the suspended rotor in the pump. The hydrodynamic performance and internal flow fields in the pump are investigated by computational fluid dynamics. The pump head flow characteristics and the efficiency-Q curves at various rotating speeds are obtained, and the detailed flow fields in the pump are determined numerically. The distribution of shear stress, including Reynolds shear stress, is studied and discussed. Also, special attention is given to the small clearance between the rotor and the pump shell where the reversed secondary flow is formed and can flush out the clearance to avoid the flow stagnations. The secondary flow as well as the magnetic bearings can reduce thrombus in the pump. To check the biocompatibility of the pump further, the hemolysis indexes of the pump are estimated on the basis of the computed results. PMID- 17117051 TI - Progress in the development of the DexAide right ventricular assist device. AB - The DexAide right ventricular assist device (RVAD) is an implantable centrifugal pump modified from the CorAide left ventricular assist device. As previously published, in vitro performance testing of the DexAide RVAD has met design criteria, and the nominal operating condition of 4 l/min and 20 mm Hg pressure rise was achieved at 2,000 rpm, with a power consumption of 1.9 watts. In vivo studies in 14 calves have demonstrated acceptable hemodynamic characteristics. The calf inflow cannula design is still evolving to minimize depositions on the cannula observed in most experiments. Fitting studies were performed in 5 cadavers and 2 patients to reconfigure the cannulae for use in humans. The design and development of external electronics have been completed for the stand-alone RVAD system, and verification tests are under way in preparation for preclinical tests. Work on the external electronics design for the biventricular assist system is ongoing. In conclusion, the initial in vitro and in vivo studies have demonstrated acceptable hemodynamic characteristics of the DexAide RVAD. The design and development of the external electronic components for the stand-alone RVAD system have been completed. The calf inflow cannula is being redesigned, and chronic in vivo tests are under way. PMID- 17117052 TI - Incorporation of an in-line filter for ultrafiltration or hemodialysis to the Abiomed BVS5000 ventricular assist device. AB - Multiple organ system dysfunction is an associated finding in patients requiring acute ventricular assist device (VAD) support. In the setting of acute renal failure, percutaneous catheters are placed for hemodialysis and/or ultrafiltration. Incorporation of an ultrafiltration or hemodialysis system in the Abiomed BVS5000 VAD circuit is a simple maneuver that eliminates the need for an additional catheter, thereby reducing the vascular and infectious complications associated with these catheters. We recommend splicing connectors into the outflow tubing of the right VAD circuit for attachment to an ultrafiltration or hemodialysis unit. This technique is a safe, simple, and reliable method by which to perform intermittent or continuous ultrafiltration or hemodialysis. PMID- 17117053 TI - A prototype of volume-controlled tidal liquid ventilator using independent piston pumps. AB - Liquid ventilation using perfluorochemicals (PFC) offers clear theoretical advantages over gas ventilation, such as decreased lung damage, recruitment of collapsed lung regions, and lavage of inflammatory debris. We present a total liquid ventilator designed to ventilate patients with completely filled lungs with a tidal volume of PFC liquid. The two independent piston pumps are volume controlled and pressure limited. Measurable pumping errors are corrected by a programmed supervisor module, which modifies the inserted or withdrawn volume. Pump independence also allows easy functional residual capacity modifications during ventilation. The bubble gas exchanger is divided into two sections such that the PFC exiting the lungs is not in contact with the PFC entering the lungs. The heating system is incorporated into the metallic base of the gas exchanger, and a heat-sink-type condenser is placed on top of the exchanger to retrieve PFC vapors. The prototype was tested on 5 healthy term newborn lambs (<5 days old). The results demonstrate the efficiency and safety of the prototype in maintaining adequate gas exchange, normal acido-basis equilibrium, and cardiovascular stability during a short, 2-hour total liquid ventilator. Airway pressure, lung volume, and ventilation scheme were maintained in the targeted range. PMID- 17117054 TI - Dialyzer fiber bundle volume during hemodialysis using large-surface dialyzers. AB - Our aim was to measure and compare fiber bundle volume (FBV) of three large surface dialyzers during chronic hemodialysis; FBV was evaluated in patients without heparin (n = 6), with tight heparinization (n = 6), and with large doses of heparin (n = 6). Each patient was treated consecutively with three dialyzers: Nephral ST500, Tricea210, and Optiflux200NR. FBV was measured hourly by ultrasound dilution in 54 sessions (n = 270). For all patients (without heparin, tight heparin, and large heparin doses) FBV did not vary significantly from 0 to 4 hours for the three types of dialyzers. There was no significant difference in aspect of filters/tubings at the end of dialysis and the presence of clots in the circuit assessed hourly. In addition, 6 other patients on hemodialysis without heparin were treated with Nephral ST500 alternatively with and without saline flushes; thus, 12 more sessions were monitored hourly for FBV (n = 60). There was no change in FBV from 0 to 4 hours between the two approaches, and the appearance of dialyzers/tubings was similar. We conclude that the FBV of large-surface dialyzers is well maintained during 4-hour chronic high-flux hemodialysis and that the loss of dialyzer surface does not explain the difference between prescribed and delivered dialysis dose in such circumstances. The membrane AN69ST does not appear more thrombogenic nor require saline flushes for heparin-free hemodialysis. PMID- 17117055 TI - Role of free hemoglobin in 8-iso prostaglandin F2-alpha synthesis in chronic renal failure and its impact on CD163-Hb scavenger receptor and on coronary artery endothelium. AB - Free hemoglobin (Hb) during autoxidation increases 8-iso-prostaglandin-F2-alpha (8-isoprostane) formation in vitro. Because 8-isoprostane and plasma Hb are elevated in chronic renal failure (CRF), we evaluated the role of Hb in this isoprostane synthesis in vivo. By monitoring correlations between Hb, haptoglobin (Hp), CD163-Hb-scavenger receptor, and 8-isoprostane that is known to induce CD163 shedding, we examined whether 8-isoprostane blocks Hb catabolism in CRF. Additionally, by studying the effect of 8-isoprostane on human coronary artery endothelium (HCAEC) in vitro and its impact on intercellular adhesion molecule-1 (ICAM-1) in vivo, we tested its role in promotion of cardiovascular events in CRF. Twenty-two never-dialyzed CRF patients and 18 control patients were screened for renal function, plasma and urine 8-isoprostane, and plasma Hb, Hp, thiobarbituric-acid-reactants (TBARS), C-reactive-protein (CRP), and soluble (s) ICAM-1 and sCD163. HCAEC exposed to 8-isoprostane were tested for ICAM-1 and apoptosis. In CRF, urine 8-isoprostane was significantly elevated and correlated with free-Hb and TBARS. The increased free-Hb, Hp, and sCD163 in CRF suggested 8 isoprostane-mediated suppression of Hb catabolism through CD163 receptor shedding. 8-Isoprostane enhanced ICAM-1 expression and apoptosis in HCAEC. CRF patients showed elevated sICAM-1. In conclusion, free-Hb, via 8-isoprostane, paradoxically blocks its own catabolism. Free-Hb and/or 8-isoprostane may intensify cardiovascular events in CRF. PMID- 17117056 TI - Device and technique for extracorporeal blood volume sequestration during hemodialysis. AB - We developed a technique for a controlled and reversible volume perturbation of the cardiovascular system during hemodialysis. The capacitance of the extracorporeal circulation was modified by an expansion bag, using separate filling and draining lines connected to postpump and prepump sections of the arterial line segment, respectively. The connection to this bag was manually operated by three-way valves. The volume sequestered into the blood bag was continuously measured by a weighing scale. Filling and draining of the expansion bag was measured in eight patients in two subsequent routine dialysis treatments. Four volume shifts were done in each treatment. Filling and draining rates and times depended on extracorporeal blood flow as well as arterial and venous line pressures. Bag inflow and outflow volumes were 215.8 +/- 28.5 ml/min and 221.6 +/ 37.0 ml/min, respectively. The total volume transiently sequestered in the bag was 479.8 +/- 61.3 ml. The duration of the whole test was 4.5 +/- 0.8 minutes. During filling, residual dialyzer blood flow was transiently reduced to 58.5 +/- 21.6 ml/min and the filtration fraction reached 30 +/- 13%. There were no adverse events such as clotting in the blood bag or in the dialyzer. The method provides rapid, reversible, and safe volume shifts between the patient and the extracorporeal circulation with the potential to elicit a hemodynamic response for characterizing the patient during each dialysis treatment. PMID- 17117057 TI - Early hemodynamic improvement is a prognostic marker in patients treated with continuous CVVHDF for acute renal failure. AB - We examined whether hemodynamic improvement after high-flow hemofiltration predicts survival in patients treated with standard continuous renal replacement therapy (CRRT). This was a prospective, observational cohort study of 169 patients, measuring the mean arterial pressure (MAP) and norepinephrine (NE) dosage before and 24 hours after CRRT. Responders were defined as having a 20% reduction in NE dosage or a 20% rise in MAP with no increase in NE, compared with nonresponders. Patients were considered to be unstable if they were receiving NE or their MAP was lower than 60 mm Hg before CRRT. Of the 169 patients, 68% were men; mean age was 53.8 years (52.7 to 54.9), with a mean Acute Physiology and Chronic Health Evaluation (APACHE) II at admission of 21.8 (21.2 to 22.3), of whom 114 were unstable at the start of CRRT. Overall mortality rate 15 days after the end of CRRT was 54.3% (57.7% in stable vs. 52.9% in unstable patients, p = NS). There were 99 responders and 70 nonresponders, the only differences being NE dosage (higher in responders, p < 0.01) and mortality rate (responders 30% vs. nonresponders 74.7%, p < 0.001). In unstable patients, mortality rate was 30% in responders versus 87% in nonresponders (p < 0.001) (72% sensitivity and 86% specificity for predicting death). Logistic regression analysis showed that the only variables associated with death were APACHE II at admission (OR, 1.06; 95% CI, 1.0 to 1.12), percent creatinine decrease (OR, 0.98; CI, 0.96 to 1.0), and lack of hemodynamic response to CRRT (OR, 7.04; CI, 3.3 to 15.02). Hemodynamic improvement after 24-hour CRRT is a strong predictor of survival. PMID- 17117058 TI - Predictors of clinical outcome in advanced heart failure patients on continuous intravenous milrinone therapy. AB - Home-based milrinone therapy (HMT) is used as a bridge to cardiac transplant (CT). The safety, efficacy, and predictors of success of HMT were assessed. Forty five patients with heart failure, referred for CT, were prospectively studied. After initial assessment, low-dose milrinone was titrated based on clinical response. Hemodynamic status was then reevaluated. Thirty-nine patients were discharged on HMT. Patients needing a left ventricular assist device (LVAD) despite milrinone (group I) and those not requiring LVAD (group II) were compared. Six of the 45 patients were ineligible for CT; 16 of 39 required LVAD as a bridge to CT despite milrinone (group I); 23 were stable on milrinone and did not require LVAD (group II). Group I was younger than group II (mean age 38.4 +/- 14.5 years vs. 57.3 +/- 5.9 years, p < 0.001). Initial acute response to intravenous milrinone [e.g., fall in the PCWP (-10.7 +/- 9.5 vs. -2.7 +/- 10.4, p = 0.02), rise in pulmonary artery oxygen saturations (16.5 +/- 8.7 vs. 7.3 +/- 10.9, p = 0.05)] was significantly better in group II than in group I. Acute hemodynamic response to milrinone predicts success of HMT as a bridge to CT. PMID- 17117059 TI - Initial experience with the development and numerical and in vitro studies of a novel low-pressure artificial right ventricle for pediatric Fontan patients. AB - The Fontan operation, an efficient palliative surgery, is performed for patients with single-ventricle pathologies. The total cavopulmonary connection is a preferred Fontan procedure in which the superior and inferior vena cava are connected to the left and right pulmonary artery. The overall goal of this work is to develop an artificial right ventricle that can be introduced into the inferior vena cava, which would act to reverse the deleterious hemodynamics in post-Fontan patients. We present the initial design and computational analysis of a micro-axial pump, designed with the particular hemodynamics of Fontan physiology in mind. Preliminary in vitro data on a prototype pump are also presented. Computational studies showed that the new design can deliver a variety of advantageous operating conditions, including decreased venous pressure through proximal suction, increased pressure rise across the pump, increased pulmonary flows, and minimal changes in superior vena cava pressures. In vitro studies on a scaled prototype showed trends similar to those seen computationally. We conclude that a micro-axial flow pump can be designed to operate efficiently within the low-pressure, low-flow environment of cavopulmonary flows. The results provide encouragement to pursue this design to for in vitro studies and animal studies. PMID- 17117060 TI - Postoperative prophylactic peritoneal dialysis in neonates and infants after complex congenital cardiac surgery. AB - Peritoneal dialysis after complex congenital cardiac surgery was introduced to a group of neonates and infants (n = 756; age, 0 to 1 year) between May 1993 and December 2005. Indications of peritoneal dialysis were determined as well as methods, prolonged dialysis, and its outcomes. Demographic characteristics, preoperative risk factors, intraoperative variables, and postoperative complications were compared in 756 cases with ages below 1 year. All cases underwent ultrafiltration during the perioperative stage. One hundred eighty-six cases (24.6% of total) required peritoneal dialysis. The cardiac pathology was transposition of great arteries in 133 cases, tetralogy of Fallot in 37, aorticopulmonary window associated with interrupted aortic arch in 4 and total anomalous pulmonary venous return in 5, and other complex pathology in 7 cases. Prolonged peritoneal dialysis was usually required in infants with low weight, with episodes of pulmonary hypertensive crisis (p < 0.05), and with preoperative renal dysfunction. No major complication was observed related to the peritoneal dialysis catheter. Of 186 patients, 23 (12.3%) had acute renal failure, and 4 of them died (2.15% of all patients underwent operation, 17.3% of those with acute renal failure). It has been demonstrated that the combination of peritoneal dialysis with perioperative ultrafiltration application was effective in providing the required postoperative negative fluid balance in especially complex congenital heart cases and affected survival positively. PMID- 17117061 TI - Microfluidic devices for continuous blood plasma separation and analysis during pediatric cardiopulmonary bypass procedures. AB - As an extension of previous work, a microfluidic device, which can separate blood plasma in a continuous, real-time fashion from a whole blood, is successfully integrated with a mock cardiopulmonary bypass circuit. The functionality of the device is demonstrated with the use of freshly harvested bovine blood. The plasma selectivities were 100% and 99.4% and the plasma separation volume percents were 18.7% and 24.5% for 26% and 37% inlet hematocrit levels, respectively. As an advanced stage of this research, a microfluidic device, which can measure the concentration of clinically relevant blood plasma protein in a continuous fashion, is being developed on the basis of fluid handling circuits coupled to fluorescent cytometric bead assays. The functionality of the device is demonstrated with the use of a biotinylated FITC solution and a streptavidin coated, 8-mum-diameter bead. The binding event between biotinylated FITC and the streptavidin bead is continuously detected within a detection window at the outlet of the device. For a known concentration (1 microg/ml) of biotinylated FITC solution, the measured fluorescent intensity is fairly constant and shows a stable gaussian distribution of the bead fluorescence intensity. It is expected that the proposed device can be used for continuous measurement of clinically relevant proteins during cardiac surgery with the cardiopulmonary bypass procedure. PMID- 17117062 TI - Subcutaneous low molecular weight heparin for management of anticoagulation in infants on excor ventricular assist device. AB - Anticoagulation in infants and children on a ventricular assist device presents particular challenges. Unfractionated heparin has poor bioavailability; it can be difficult to achieve a stable anticoagulant effect; and, in the long-term, there is a risk of osteopenia. Long-term warfarin can be difficult to manage in infants on formula milk with vitamin K supplementation. We review our recent experience with subcutaneous low molecular weight heparin. Two patients received a left ventricular assist device (Excor, Berlin Heart AG) as a bridge to transplantation. Initial anticoagulation consisted of unfractionated heparin infusion beginning 6 hours after implantation to maintain an activated partial thromboplastin time of 70 seconds, checked every 4 to 6 hours. Platelet count (aim >80,000/microl) and thromboelastography were assessed daily. Antithrombin required substitution to maintain levels >70 IU/dl. To optimize anticoagulation, both infants were switched to subcutaneous low molecular weight heparin twice daily aiming for an anti-Xa activity between 0.5 and 1.0 IU/ml. Aspirin was added on day 4, checking platelet aggregation every 2 to 4 days, aiming at arachidonic acid stimulated aggregation 10% to 30% of baseline, collagen 100% of baseline. Dipyridamole was added once stability was reached if platelets count exceeded 150,000/microl. There were no clinical thromboembolic or bleeding events. Both patients had successful transplantation. PMID- 17117063 TI - Modified CPB circuit for postoperative rescue of high-risk patients following cardiac repair: are we keeping safe? AB - Extracorporeal membrane oxygenation (ECMO) is commonly used to treat postcardiotomy cardiopulmonary dysfunction in small children. System readiness, need for additional blood products, and exposure to new surfaces are important considerations, particularly when used for resuscitation. We reviewed our experience with a cardiopulmonary bypass (CPB) system modified to provide extended circulatory support system after surgery in patients considered at high risk. When not used in the operating room, the system was recirculated for 24 hours. Before being discarded, blood samples were obtained for activated clotting time, arterial blood gas, and blood cultures from 10 circuits. Between January 2004, and December 2005, 44 patients underwent cardiac repair using this CPB system. ECMO support was initiated in the operating room in 8 patients, and six circuits were used after patient arrival in the intensive care unit. Blood sampling after 24 hours on standby circuits revealed acceptable values for pH, Pao2, hematocrit, ionized calcium, potassium level, and ACT. All blood cultures were negative at 5 days. Survival for patients who received a circuit on standby was 64%.This modified cardiopulmonary circuit can be transformed into a simple, safe, and effective ECMO support system. Deployment of a CPB circuit previously used for cardiac repair has many advantages and maximizes utilization of resources. PMID- 17117064 TI - Quantification of perfusion modes in terms of surplus hemodynamic energy levels in a simulated pediatric CPB model. AB - The objective of this investigation was to compare pulsatile versus nonpulsatile perfusion modes in terms of surplus hemodynamic energy (SHE) levels during cardiopulmonary bypass (CPB) in a simulated neonatal model. The extracorporeal circuit consisted of a Jostra HL-20 heart-lung machine (for both pulsatile and nonpulsatile modes of perfusion), a Capiox Baby RX hollow-fiber membrane oxygenator, a Capiox pediatric arterial filter, 5 feet of arterial tubing and 6 feet of venous tubing with a quarter-inch diameter. The circuit was primed with a lactated Ringers solution. The systemic resistance of a pseudo-patient (mean weight, 3 kg) was simulated by placing a clamp at the end of the arterial line. The pseudo-patient was subjected to five pump flow rates in the 400 to 800 ml/min range. During pulsatile perfusion, the pump rate was kept constant at 120 bpm. Pressure waveforms were recorded at the preoxygenator, postoxygenator, and preaortic cannula sites. SHE was calculated by use of the following formula {SHE (ergs/cm) = 1,332 [((integral fpdt) / (integral fdt)) - Mean Arterial Pressure]} (f = pump flow and p = pressure). A total of 60 experiments were performed (n = 6 for nonpulsatile and n = 6 for pulsatile) at each of the five flow rates. A linear mixed-effects model, which accounts for the correlation among repeated measurements, was fit to the data to assess differences in SHE between flows, pumps, and sites. The Tukey multiple comparison procedure was used to adjust p values for post hoc pairwise comparisons. With a pump flow rate of 400 ml/min, pulsatile flow generated significantly higher surplus hemodynamic energy levels at the preoxygenator site (23,421 +/- 2,068 ergs/cm vs. 4,154 +/- 331 ergs/cm, p < 0.0001), the postoxygenator site (18,784 +/- 1,557 ergs/cm vs. 3,383 +/- 317 ergs/cm, p < 0.0001), and the precannula site (6,324 +/- 772 ergs/cm vs. 1,320 +/ 91 ergs/cm, p < 0.0001), compared with the nonpulsatile group. Pulsatile flow produced higher SHE levels at all other pump flow rates. The Jostra HL-20 roller pump generated significantly higher SHE levels in the pulsatile mode when compared with the nonpulsatile mode at all five pump flow rates. PMID- 17117065 TI - Patterns of red marrow in the adult femur. AB - PURPOSE: Conversion of red marrow (RM) to fatty marrow in the skeleton of the lower extremities begins at the distal end, ie, feet, and progresses proximally with distal bone marrow (ie, tibia) being converted more rapidly than proximal bone marrow (ie, femur). However, in an individual long bone, conversion begins in the diaphysis and progresses both distally and proximally (more rapidly toward the distal side). In a normal adult's femur, RM is present in the proximal one third or less. Reconversion of fatty marrow to RM is reported to occur in the reverse order of conversion. We assessed the frequency of various patterns of RM in the adult femur on In-111 leukocyte scans for a better understanding of the bone marrow regeneration process in individual long bones. METHODS: The patterns of marrow activity in the femur shown on In-111 leukocyte scans performed in 354 adults were divided into a) RM limited to the proximal one third or less, b) to the proximal two thirds, c) to the proximal one third and distal one third with no activity in the middle shaft, and d) in the entire femur. RESULTS: There were 207 patients with pattern A, 91 pattern B, 14 pattern C, and 42 pattern D. CONCLUSIONS: A considerably higher number of adults showed pattern B than pattern C. This suggests that regeneration of diaphyseal marrow precedes that of the distal marrow in an individual long bone or possibly that conversion of the latter precedes the former, which is different from that proposed in the literature. PMID- 17117066 TI - Characteristics of skeletal stress fractures in female military recruits of the Israel defense forces on bone scintigraphy. AB - AIM: Clinical surveys on stress fractures (SF) in female military recruits are scarce. The aim of this study was to characterize the scintigraphic findings and classify the distribution and pattern of SF in a group of female recruits of the Israel Defense Forces (IDF). MATERIALS AND METHODS: The bone scans of 146 female recruits (age range, 19-20.6 years) with suspected SF were assessed retrospectively. The SF lesions were classified qualitatively into 4 grades of bone response according to the classification criteria introduced by Zwas et al. SF location and distribution were analyzed, and in several cases, the abnormalities were correlated radiographically. RESULTS: One hundred forty-six female recruits were examined, of which 93 (64%) had bone scan findings of SF with a total of 247 SF. Forty-eight patients (with or without SF) had shin splints, 32 had thigh splints, and 34 had normal scans. Several SF were detected in sites that were not clinically suspected. Thirty-nine percent of the SF were located in the feet (tarsal bones 22.7%, metatarsal 16.2%), 36.8% in tibiae (predominantly in the midthird), 15.7% in femurs, 6.5% in the pubic and sacroiliac regions, and 2% in the fibula. SF in the tibiae and femurs were mainly located in the posterior aspect of the medial cortical region. Forty-nine percent of the patients had bilateral SF. The SF were classified on a 4-grade scale: 41.3% were grade I, 37.2% grade II, 15.8% grade III, and 5.7% grade IV. Thirty three percent of the patients had one site of SF, 31% had 2 sites, 7.5% had 3 sites, 12% had 4 sites, 7.5% had 5 sites, and 9% had more than 5 sites of SF. Different grades of lesions were often found in the same patient. Sixty-five percent of the patients had SF in the feet, 59% in the tibiae, 26% in the femurs, 14% in the pubic or sacroiliac regions, and 5% in the fibula. Radiography was performed in 15% of the patients. Only one patient had a positive finding on radiography. CONCLUSIONS: This study shows that lower leg SF are not uncommon in female military recruits. We clearly distinguished between SF and shin splints, which have important clinical consequences on treatment. Most of the SF were mild (grade I and II) located in the feet and tibiae. This study supports the experience gained in other studies, and emphasizes the significant number of pubic and sacroiliac SF in female soldiers, which is significantly higher in comparison with previous reports on male soldiers. PMID- 17117067 TI - MRI-negative, bone scintigram-positive in early osteonecrosis of the knees. AB - Nontraumatic avascular necrosis (AVN) of bone is a well-reported complication of glucocorticoid therapy for immunologic and malignant disease. We present the case of a 13-year-old girl with no history of trauma who presented with a 5-day history of increasing pain in both knees after cord blood transplantation for acute lymphoblastic leukemia. Plain film and magnetic resonance imaging (MRI) were reported as normal. Bone scintigraphy revealed evidence of bilateral avascular necrosis in the distal femora. MRI subsequently became abnormal several weeks later. The case illustrates the natural history of AVN, in which changes that are detected by MRI can take several weeks to develop. The scintigraphic findings influenced early management of the condition. PMID- 17117068 TI - F-18 FDG PET/CT evaluation of osseous and soft tissue sarcomas. AB - INTRODUCTION: Osseous and soft tissue sarcomas (OSTS) represent a histologic heterogeneous group of malignant tumors. Most of the current clinical data on the role of F-18 FDG PET in sarcomas come from patients studied with dedicated PET and less frequently with hardware fusion PET/CT. Therefore, we were prompted to review our experience with F-18 FDG PET/CT in OSTS. METHODS: This is a retrospective study (January 2003-December 2005) of 44 patients with histologic diagnoses of OSTS who had F-18 FDG PET/CT at our institution. The group included 22 men and 22 women with an age range of 2 of 84 years (average, 37 +/- 20.2 years). The administered doses of F-18 FDG range 4.1 to 19.5 mCi (average, 14.3 +/- 3 mCi). Reinterpretation of the imaging studies for accuracy and data analysis from medical records was performed. RESULTS: The sensitivity and specificity of combined F-18 FDG PET/CT were 100% (95% confidence interval [CI] = 75.7-100) and 93.3% (95% CI = 78.7-98.1) for the primary OSTS, and 80% (95% CI = 58.4-91.9) and 86.4% (95% CI = 66.7-95.2) for metastases. When interpreted separately, CT outperformed PET for pulmonary metastases detection: CT was 76.5% sensitive and 88% specific, whereas PET was only 57.1% sensitive but 96.4% specific. For detection of other metastases, CT was 82.3% sensitive and 76% specific, with PET demonstrating 78.6% sensitivity and 92.8% specificity. CONCLUSION: Relatively similar results (except better specificity for PET and PET/CT) were noted when examining the rate of metastases detection, excluding pulmonary lesions. However, CT had a better detection rate for pulmonary metastases when compared with PET alone. A negative PET scan in the presence of suspicious CT findings in the chest cannot reliably exclude pulmonary metastases from OSTS. PMID- 17117069 TI - Usefulness of scintimammography with tc-99m MIBI in clinical practice. AB - Radiographic mammography (MM) is routinely used to diagnose breast cancer. MM has a number of well-known limitations, especially in cases of a dense or dysplastic breast. Scintimammography (SM) with Tc-99m MIBI has been successfully used as a useful complement to MM. The authors report a case of a 57-year-old woman with MM with tiny calcifications in the left breast, which were classified by the radiologist as probably benign lesions. SM with Tc-99m MIBI showed a focal area of increased uptake in the upper outer quadrant of the left breast. On the basis of SM results, an excisional biopsy was performed and pathologic examination revealed infiltrating ductal carcinoma. PMID- 17117070 TI - Diagnosis of dementia using perfusion SPECT imaging at the patient's initial visit to a cognitive disorder clinic. AB - PURPOSE: This study was performed to evaluate the diagnostic accuracy of brain perfusion SPECT in patients visiting the cognitive disorder clinic for initial evaluation using 3D-SSP compared with using standard transaxial section. MATERIALS AND METHODS: Standard transaxial section displays and 3D-SSP z-score maps obtained after administration of Tc-99m ECD or I-123 IMP were randomly interpreted in 315 patients who visited initially to the cognitive disorder clinic (age 46-88 years; 162 women, 153 men). Sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy were calculated and receiver operating characteristic (ROC) analysis was performed. RESULTS: One hundred thirty-seven patients were clinically diagnosed with Alzheimer disease and 178 patients were diagnosed with other disorders and age associated cognitive decline. Sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy calculated using standard transaxial display were 61%, 70%, 61%, 70%, and 66%, respectively, and using the 3D-SSP z-score map were 90%, 74%, 73%, 90%, and 81%, respectively. Particularly, the sensitivity value improved in mild cases compared with severe cases using 3D-SSP. Diagnostic performance with 3D-SSP was superior in both mild dementia (Az = 0.64 [section] vs 0.81 [3D-SSP], P = 0.001) and severe dementia (Az = 0.75 [section] vs 0.90 [3D-SSP], P = 0.002). CONCLUSIONS: Brain perfusion SPECT was useful for diagnosis in patients who come to the cognitive disorder clinic for initial evaluation using 3D-SSP. PMID- 17117071 TI - Evaluation of primary brain tumors with FLT-PET: usefulness and limitations. AB - PURPOSE OF THE REPORT: The purpose of this report was to investigate the potential of positron emission tomography using F-18 fluorodeoxythymidine (FLT PET) in evaluating primary brain tumors. MATERIALS AND METHODS: FLT-PET was performed in 25 patients with primary brain tumors. FLT uptake in the lesion was semiquantitatively evaluated by measuring the maximal standardized uptake value (SUVmax) and the tumor-to-normal tissue ratio (TNR). SUVmax and TNR were compared with the histologic grade and the expression of the proliferation marker (Ki-67). RESULTS: FLT uptake in normal brain parenchyma was very low, resulting in the visualization of brain tumors with high contrast. Both SUVmax and TNR significantly correlated with the malignant grade of brain gliomas, in which high SUVmax/TNR was obtained for high-grade gliomas. Patients with primary lymphoma also showed SUVmax/TNR equivalent to glioblastoma. There was a positive correlation between SUVmax/TNR and the Ki-67 index. In contrast, spuriously high SUVmax and TNR were obtained in 3 of 6 patients with suspected recurrent tumors (2 patients with recurrent grade 2 glioma and one patient with postoperative granuloma), all of which showed lesion enhancement on MRI after Gd administration. CONCLUSIONS: FLT-PET can be used to evaluate the malignant grade and proliferation activity of primary brain tumors, especially malignant brain tumors. However, the presence of benign lesions showing blood-brain barrier disruption cannot be distinguished from malignant tumors and needs to be carefully evaluated. PMID- 17117072 TI - Endometrial and ovarian F-18 FDG uptake in serial PET studies and the value of delayed imaging for differentiation. AB - PURPOSE: This study was designed to characterize the physiological endometrial and ovarian F-18 FDG uptake through analysis of clinical patients with serial PET follow ups (group 1) or who underwent delayed imaging (group 2). METHODS: Group 1 had 24 patients (14 premenopausal and 10 postmenopausal). Each patient underwent 2 to 4 serial FDG PET studies and summed to 57 studies. Group 2 included 15 premenopausal patients with delayed imaging 3 hours after injection. Ten of them showed endometrial uptake, and 8 showed intense uptake in 10 ovaries. Visual and semiquantitative methods were applied for analysis. RESULTS: By analysis of serial PET studies, endometrial and ovarian uptake showed some specific characteristics. Surrounded by a relatively low-uptake uterine wall, the inverted cone-shaped endometrium showed 2 peaks of uptake, one in early menstruation and the other in midcycle. The ovarian uptake was more prominent in the midcycle, and the foci of uptake had an ovoid shape and located at the left and/or right side superior-posterior to the bladder. For the postmenopausal and amenorrhea women (35 studies), only one study in a woman within 1 year of menopause showed mild endometrial and ovarian uptake. Compared with the imaging 1 hour after injection, the standardized uptake value increased in 2 of 10 endometria and 4 of 10 ovaries in the delayed imaging. CONCLUSION: Physiological endometrial and ovarian uptake can be identified on PET alone by their specific shape and position as well as their relation to the menstrual state, whereas delayed imaging was of little value for differentiation in this preliminary study. PMID- 17117073 TI - Fluorine-18 fluorothymidine: a new positron emission radioisotope for renal tumors. AB - Fluorine-18 fluorothymidine (F-18 FLT) is a radioisotope based on the nucleic acid thymidine and has emerged as an important tracer that mirrors cellular proliferation in positron emission tomography (PET) studies. Early studies in human tumors have been promising. However, imaging of renal tumors using F-18 FLT PET studies has not previously been described. In this report, a difficult case of renal transitional cell carcinoma in a longstanding cyst was clearly delineated using F-18 FLT. Importantly, the study was able to guide clinicians toward appropriate surgical management. The use of such tracers may herald a new era in renal tumor imaging. PMID- 17117074 TI - Low-dose P32 therapy in essential thrombocythemia. AB - We present a case of an 85-year-old woman with medically refractory essential thrombocythemia and subsequent venous thrombosis. She received conservative phosphorus-32 sodium phosphate therapy for 3 mCi, approximately half the usual dose. One month later, she received a second intravenous phosphorus-32 treatment of 3.5 mCi. She responded successfully to both treatments with drops in her platelet count and experienced no adverse effects. Our case is noteworthy in the effectiveness from a conservative dose while avoiding hematologic complications. PMID- 17117075 TI - Positive correlation between standardized uptake values of FDG uptake in the stomach and the value of the C-13 urea breath test. AB - PURPOSE: The distribution of FDG uptake in the stomach is variable. Gastritis is one of the causes of elevated FDG uptake. Helicobacter pylori (HP) infection was found in approximately 80% of patients with peptic ulcer and 30% to 60% of symptomatic gastritis. The C-13 urea breath test (UBT) is one of the gold standard tools in diagnosis of HP infection. The aim of this study was to assess the correlation between standardized uptake values (SUVs) of FDG uptake in the stomach and the value of the C-13 urea breath test (UBT). METHODS: Sixteen patients, referred from the department of community medicine, for FDG PET for health examination were chronologically enrolled for detection of HP infection by using the C-13 UBT within 1 week. The maximal SUVs of FDG uptake in the gastric wall as well as the value of the C-13 UBT were measured. The correlation between maximal SUVs of FDG uptake in the stomach and the value of the C-13 UBT was observed. RESULTS: We took the results of C-13 UBT as the final diagnosis of HP infection. Of 16 patients, 5 were HP-infected and 11 were uninfected. Standardized uptake values of FDG uptake in the gastric wall were strong positively correlated with the value of the C-13 UBT in this retrospective study (P < 0.001). CONCLUSION: Except for gastric cancer, HP infection has to be concerned when high SUVs of FDG uptake in the stomach are noted. PMID- 17117076 TI - The use of periareolar intradermal Tc-99m tin colloid and peritumoral intraparenchymal isosulfan blue dye injections for determination of the sentinel lymph node. AB - PURPOSE: The purpose of the present study was to evaluate the use of lymphoscintigraphy, blue dye, and gamma probe detection methods for determination of the sentinel lymph node (SLN) using both periareolar intradermal injection of Tc-99m tin colloid and peritumoral intraparenchymal injection of isosulfan blue dye. METHODS: One hundred patients with T1-2 breast cancer and clinically negative nodes were enrolled in the present study. The study was composed of 2 groups. Backup axillary lymph node dissection (ALND) was mandatory in group 1 (20 patients) regardless of their lymph node status. In group 2 (80 patients), complete ALND was performed when intraoperative frozen section analysis of SLN revealed metastases. Otherwise, only SLN biopsy was performed without ALND. One day before surgery, Tc-99m tin colloid was injected at 4 periareolar sites intradermally. Lymphoscintigraphy was performed 1 to 2 hours after injection of the radiocolloid. Twenty minutes before surgery, isosulfan blue dye was injected into parenchyma surrounding the tumor or the biopsy cavity. RESULTS: The detection rates of SLN and false-negative rate of lymphoscintigraphy, blue dye, and gamma probe detection were 85%, 95% 100%, and 0% in group 1, 91%, 87%, and 95% in group 2, respectively. Detection rate by the combination of blue dye and radio tracer was 98%. CONCLUSIONS: According to the results of our study, we conclude that perioareolar intradermal injection of Tc-99m tin colloid combined with peritumoral intraparenchymal injection of blue dye is an accurate and easy method of locating the sentinel node with very high detection rates. It is recommended that the combination of all methods such as lymphoscintigraphy, blue dye, and gamma probe application will increase the success rate of SLN detection in patients with breast cancer. PMID- 17117077 TI - Fused SPECT/CT imaging of Peri-iliopsoas infection using Indium-111-labeled leukocytes. AB - Nuclear imaging with In-111-labeled leukocytes has become an instrumental tool in localizing sites of infection and is superior to Ga-67 in localizing abdominal and pelvic abscesses resulting from absence of a normal bowel excretory pathway. Labeled white blood cells (WBCs) localize at sites of infection through diapedesis, chemotaxis, and enhanced vascular permeability and can thus be used to identify infection. The accuracy of this functional imaging modality can be enhanced by fusing SPECT images of labeled WBC with CT images that provide anatomic detail to facilitate reading as illustrated in the case described. PMID- 17117078 TI - FDG PET/CT flip flop phenomenon in treated lymphoma of bone. AB - A 47-year-old man with primary large B-cell lymphoma of bone underwent an F-18 fluorodeoxyglucose positron emission tomography/computed tomography (FDG PET/CT) scan for staging. The study demonstrated multiple areas of uptake consistent with osseous lymphoma. After multiple cycles of chemotherapy, a follow-up study demonstrated a dramatic flip flop appearance in which the previously noted areas of osseous lymphoma were photopenic and normal marrow appeared to have increased activity. This flip flop appearance could incorrectly suggest lymphomatous infiltration of normal marrow. PMID- 17117079 TI - Tc-99m pentavalent DMSA scintigraphy in myelofibrosis detection. AB - In a 62-year-old man with medullary carcinoma of the thyroid, a postoperative Tc 99m dimercaptosuccinic acid [(V) DMSA] study was requested. In the Tc-99m (V) DMSA scan. no abnormalities, indicating local recurrence or metastatic disease, were observed. However, there was increased uptake in the spleen and liver and significantly diffusely increased uptake in the bone marrow. The patient also had a history of myelofibrosis and these findings appear to have been the result of this pathology. PMID- 17117080 TI - Bilateral Paget disease of the calcaneus diagnosed by conventional bone scintigraphy. AB - An 85-year-old woman who had an invasive ductal carcinoma of the breast and elevated alkaline phosphatases (420 U/L) was referred for Tc-99m MDP bone scintigraphy for initial evaluation of skeletal metastases. Bone symptoms were limited to chronic pain in both knees. A bone scan revealed pagetoid findings in both calcanei. Feet and toes on plain x-rays of the calcaneus were unremarkable. Findings were stable in the 2-year follow up, excluding other potential diagnosis as extensive metastases or stress fractures. PMID- 17117081 TI - Multiple metastases to skeletal muscle from carcinoma of the esophagus detected by FDG PET-CT imaging. AB - A 67-year-old woman was treated with neoadjuvant chemotherapy, esophagectomy, and subsequent radiotherapy for T3N1 poorly differentiated adenocarcinoma of the esophagus. Five months after surgery, a routine follow-up CT demonstrated a 1.2 cm soft tissue mass in the posterior mediastinum suspicious for local recurrence. An FDG-PET/CT study confirmed tumor in the posterior mediastinum and also showed focal areas of increased tracer uptake within several muscles. Skeletal muscle is one of the most unusual sites of metastatic disease, although it is probable that the more frequent use of FDG-PET imaging will lead to an increase in the detection of such lesions. PMID- 17117082 TI - Gallbladder metastasis from malignant melanoma: diagnosis with FDG PET/CT. AB - Melanoma with metastasis to the gallbladder is sometimes seen on autopsy but is rarely seen in living patients, in part because it is often asymptomatic. A 67 year-old man with a history of malignant melanoma in situ underwent an F-18 FDG PET/CT scan, which showed a gallbladder focus (SUV 16.9). Four months later, on the repeat FDG PET/CT scan, a new lesion in the gallbladder was noted. Laparoscopic cholecystectomy was done and histopathologic findings were consistent with gallbladder metastasis of melanoma. PET/CT detects metastasis at unusual sites accurately and is helpful in correct staging and management of patients with melanoma. PMID- 17117083 TI - Meckel's scan: pitfall in patients with active small bowel bleeding. AB - Potential for a false-positive Meckel's scan is highlighted in this patient with intermittent, low-volume small bowel bleeding. Meckel's scan showed late appearance of focal activity mimicking a positive diverticulum finding. This was secondary to extravasated tracer during a coincidental bleeding as shown by a Tc 99m tagged red blood cell scan obtained 2 days earlier. Inaccurate diagnosis of Meckel diverticulum was avoided by: 1) noticing the late appearance in relationship to the gastric uptake, and 2) showing motility of the focus on delayed dynamic imaging that confirmed transit of extravasated tracer along the small bowel with peristalsis. Subsequent capsule endoscopy showed a bleeding ulcer in the ileum. PMID- 17117084 TI - Incidental detection of previously undiagnosed primary breast carcinoma with a gallium-67 scan performed for evaluation of occult infection. AB - A 63-year-old female patient with a recent left posterior cerebral artery infarction and pyelonephritis presented with persistent fevers despite adequate intravenous antibiotic therapy. Whole-body gallium-67 scintigraphy was performed to evaluate for occult infection. Tracer accumulated bilaterally in the kidneys consistent with the previously diagnosed pyelonephritis. Relatively intense uptake was also noted in the area of a known occipitoparietal infarct. Incidentally noted was an abnormal focus of increased tracer uptake in the retroareolar region of the right breast. Subsequent guided core biopsy of the right breast mass demonstrated a previously undiagnosed invasive ductal carcinoma. PMID- 17117085 TI - F-18 FDG PET scan findings in a case of carcinoma of the breast with a rare site of metastases to the gingival region. AB - Malignant tumors of the breast have an inherent potential to metastasize more often to the regional lymph nodes. It is rare to find a metastasis to the oral region from a primary in the breast, but when this does occur, it usually involves the jawbones rather than the soft tissues. A 33-year-old premenopausal woman, a diagnosed case of locally advanced right breast carcinoma, underwent right modified radical mastectomy followed by chemotherapy as per the institutional protocol. She presented after 2 years with an exophytic growth in the upper alveolar region of the oral cavity. Biopsy indicated gingival metastasis from a poorly differentiated adenocarcinoma of the breast. She was referred for F-18 FDG PET scan to evaluate the disease status before planning radiotherapy to the gingival metastasis. F-18 FDG PET scan was done after intravenous injection of 370 MBq (10 mCi) of tracer. Whole-body PET images were reconstructed in iterative algorithm (OSEM). Whole-body F-18 FDG PET scan showed hypermetabolic foci in the midmaxillary region (SUV max: 6.5), upper end of the right humerus, a large hypermetabolic area in the upper zone of the right lung with contiguous hilar node involvement on the right side of the lung, and an area of intense hypermetabolic activity in the left acetabular and ischial region. The present case demonstrates a rare site of metastasis in the oral region from carcinoma of the breast. PMID- 17117086 TI - Multimodality imaging of malignant pheochromocytoma. AB - Pheochromocytomas offer the opportunity to explore multiple pathophysiological mechanisms through functional imaging. MIBG scintigraphy and PET scanning with tracers of the sympathetic nervous system are based on uptake of catecholamines and catecholamine-like compounds by hNET, the human norepinephrine transporter. In-111 pentetreotide scanning involves the imaging of somatostatin receptors on the cellular surface of tissues. FDG PET scanning examines the transport and incorporation of FDG into cells. We present a patient with malignant pheochromocytoma who underwent multitracer imaging to characterize the tumor and probe its pathophysiology to direct a therapeutic approach. This case underscores the inherent difficulties in the diagnosis and localization of malignant pheochromocytomas. Multiple approaches to functional and anatomic imaging may be required to fully delineate the extent of disease and similarly to direct radionuclide-based therapy. PMID- 17117087 TI - Parietal lobe epileptic focus identified on SPECT-MRI fusion imaging in a case of epilepsia partialis continua. AB - Epilepsia partialis continua, or "Kozhevnikov syndrome," is a rare condition characterized by persistent localized motor seizures usually localized in a distant limb. It is most often seen in children under 16 years old without gender preference. We report a 12-year-old girl with epilepsia partialis continua who presented to the emergency room after the onset of seizures. Routine electroencephalography (EEG) was performed with no clear ictal localization. Magnetic resonance imaging (MRI) was unremarkable. Ictal single photon emission computed tomography (SPECT) images with Tc-99m exametazime fused with MRI images revealed a parietal lobe epileptic focus within the superior segment of the postcentral gyrus. This case illustrates a rare disorder with an unusual epileptic localization identified by SPECT-MRI fusion. PMID- 17117088 TI - Exercise acutely increases renal transit time of Tc-99m mercaptoacetyltriglycine (MAG3) in a post-liver transplant patient. AB - This case demonstrates the effect of exercise on the clearance of Tc-99m MAG3 in a patient with renal insufficiency status post-liver transplant. Even after furosemide administration, the tracer was retained in the kidneys after exercise. This is in contrast to normal clearance demonstrated on a baseline study performed 3 days previously. PMID- 17117089 TI - Focal F-18 FDG uptake mimicking malignant gastric localizations disappearing after water ingestion on PET/CT images. AB - Diffuse, increased gastric wall F-18 FDG uptake is widely observed during PET/CT examinations, frequently unrelated to malignant findings, but simply caused by inflammatory disease, physiological emptying, or visceral thickening. Hence, elevated F-18 FDG gastric uptake can lead to equivocal misinterpretation, especially in patients with known gastric malignant disease, at posttherapy reevaluation. Gastric wall contraction can increase F-18 FDG uptake, especially for a remnant stomach, increasing the percentage of false-positive results with a direct impact on therapeutic management. One field PET/CT acquisition centered on the hypochondrial regions a few minutes after water ingestion should be performed routinely if standard images are doubtful (increased tracer uptake and visceral thickening) to differentiate benign from malignant uptake. PMID- 17117090 TI - Current readings in nuclear medicine. PMID- 17117091 TI - Cosmetic lower eyelid blepharoplasty with fat repositioning via intra-SOOF dissection: surgical technique and initial outcomes. AB - PURPOSE: To describe a novel surgical technique for lower eyelid blepharoplasty with fat repositioning in an intrasuborbicularis oculi fat (intra-SOOF) plane. METHODS: Description of surgical technique and outcome of a retrospective, interventional case series of 66 consecutive patients who underwent surgery. RESULTS: Sixty-six patients with an average age of 56 years underwent bilateral transconjunctival lower eyelid blepharoplasty with fat repositioning in an intra SOOF plane. Surgery was most commonly done in an office-based operating or procedure room under local anesthetic only. Patients were observed for an average of 6 months. Complications and revisions were few. Surgical outcomes and subjective patient satisfaction rate were excellent. CONCLUSIONS: Lower eyelid fat repositioning blepharoplasty in an intra-SOOF plane is an alternative technique to address the tear trough deformity. This technique provides excellent surgical results and high patient satisfaction. Convenient for both patient and surgeon, this technique is readily performed in an office-based procedure room under local anesthetic. Further follow-up will determine long-term outcomes. PMID- 17117092 TI - Use of the pericranial flap in medial canthal reconstruction: another application for this versatile flap. AB - PURPOSE: To describe the use and outcomes of a versatile surgical technique in the reconstruction of deep soft tissue and bony defects of the medial canthus. METHODS: A retrospective review of consecutive cases requiring reconstruction of medial canthal defects involving loss of periosteum or bone by a median forehead pericranial flap and full-thickness skin grafting in a tertiary referral hospital setting. Two techniques were used: an open technique, using a midline forehead incision; and an endoscopic technique, using 2 incisions behind the hairline. RESULTS: Twenty-one cases were identified: 19 open and 2 endoscopic. The average length of follow-up was 13 months (range, 6-50 months). Ten cases required additional oculoplastic procedures including local periosteal flaps and mucous membrane grafts. Two cases (10%) had complete flap failure; one of these was caused by infection. Five (24%) had partial (< 50%) skin graft necrosis. Two cases (10%) have required further surgery. CONCLUSIONS: Our experience shows the pericranial flap to be versatile, robust, and easy to manipulate, offering advantages over alternative techniques when used for the repair of deep medial canthal defects. It is a valuable reconstructive technique that can yield good cosmetic and functional results. PMID- 17117093 TI - Combined inferior and medial surgical approaches and overlapping thin implants for orbital floor and medial wall fractures. AB - PURPOSE: To evaluate the clinical outcome of combined orbital floor and medial wall fracture repair using overlapping thin implants. METHODS: A retrospective chart review was conducted on 19 fracture repairs performed by one surgeon using thin overlapping implants inserted through transcaruncular and inferior fornix incisions. RESULTS: Nineteen combined medial wall and floor fractures were repaired in 18 patients (14 male, 4 female). The median age was 37 years (range, 13-60 years). The median trauma-to-surgery interval was 9 days (range, 1 day to 6 years). Two overlapped implants (0.2-0.4 mm) were used in 12 fracture repairs, and 3 overlapped implants (0.2-0.3 mm) were used in 7 fracture repairs. Materials included prepunched nylon foil in 18 fractures and, in one fracture, high-density polyethylene. Follow-up time ranged from 1 month to 53 months (median, 7 months). No complications occurred during the postoperative follow-up period. Clinically significant enophthalmos was not observed after treatment. CONCLUSION: The repair of combined orbital floor and medial wall fractures using thin overlapping implants inserted through transcaruncular and inferior transconjunctival incisions is a safe, effective, and cosmetically sensitive surgical technique. PMID- 17117094 TI - New corticosteroid-eluting porous polyethylene implant for the management of lower eyelid retraction: a pilot study. AB - PURPOSE: Lower eyelid retraction after trauma presents a challenging management problem. We postulated that a porous polyethylene (pPE) eyelid spacer coated with a polyvinyl alcohol (PVA) and triamcinolone acetonide (TA) matrix could deliver corticosteroid locally over extended periods and modulate inflammation and scar formation. We designed a pPE corticosteroid-eluting implant and evaluated its characteristics in vitro and in vivo. METHODS: The release characteristics of pPE implants coated with a PVA/TA matrix of low, intermediate, and high doses of TA were studied in vitro. The implants were then placed in the posterior lamella of lower eyelids of Dutch Belted rabbits for 12 weeks. Clinical events were recorded and eyelids were examined for gross and histologic features, including capsular thickness and degree of vascularity, fibrovascular ingrowth, and inflammatory response. RESULTS: In vitro, implants coated with the intermediate and high doses of TA released the drug at a steady rate for at least 78 days. In rabbits, the PVA and PVA/TA coating prevented fibrovascular ingrowth, except where breaks in the PVA/TA coat were present. Implants with PVA/TA coating demonstrated less inflammation and capsule vascularity. An inverse correlation between TA dose and capsule thickness was noted. CONCLUSIONS: We describe a novel drug-release pPE eyelid implant. The corticosteroid-eluting implant demonstrated antiangiogenic and anti-inflammatory properties, which could prove beneficial in the treatment of lower eyelid retraction. PMID- 17117095 TI - Microscopic anatomy of Asian lower eyelids. AB - PURPOSE: To elucidate the microscopic anatomy of the Asian lower eyelid. METHODS: Specimens (full-thickness sections of lower eyelids from 19 postmortem lower eyelids) from 11 Asians aged 73 to 96 years at death were fixed in 10% buffered formalin and microscopically examined. After pretreatment, sagittal sliced sections of the central part were stained with Masson trichrome. RESULTS: The distinct junction of the orbital septum to the capsulopalpebral fascia (CPF) was confirmed in 7 eyelids in which orbital septum was clearly stained, with an average distance from the tarsus to the junction of 2.38 mm. The other 12 eyelids did not show a distinct junction, and the orbital septum was poorly defined anteriorly and indistinct posteriorly. There was a distinct layer between the orbicularis oculi muscle and the orbital septum. The inferior and the posterior attachments of the CPF to the tarsus were seen in all eyelids. Seventeen of the 19 eyelids had attachment of the CPF on the anterior aspect of the tarsus, from which an extension of the CPF through the pretarsal orbicularis oculi muscle was observed. All eyelids had anterior extension of the CPF through the preseptal orbicularis oculi muscle, which was overridden on the pretarsal orbicularis oculi muscle. CONCLUSIONS: The microscopic findings of Asian lower eyelids, especially fascial components, were mostly similar to those of non-Asian eyelids, but differences existed in higher or indistinct septum fusion, anterior and superior orbital fat projection, and the overriding of the preseptal orbicularis oculi muscle. PMID- 17117096 TI - A modified posterior approach for upper eyelid retraction. AB - PURPOSE: To describe and review a graded technique for lowering the upper eyelid from a posterior approach, recessing or resecting Muller muscle and levator aponeurosis but leaving a central pillar of Muller muscle intact, occasionally including a temporal tarsorrhaphy or superior tarsal strip where needed, and performing tissue dissection with a hot-wire cautery instrument. METHODS: A 10 year retrospective chart review was performed. Where follow-up was less than 6 months, telephone interviews were conducted to assess patient satisfaction with the procedure. Statistical analysis was performed using an unpaired t test. RESULTS: Ninety-nine patients (161 eyelids, 62 bilateral and 37 unilateral) with a mean age of 47 years (range, 21-82 years) were studied. The mean follow-up period was 61 months. The mean preoperative and postoperative margin reflex distances (MRD1) were 7.3 mm (range, 4.5-10 mm) and 4.3 mm (range, 2-7 mm), respectively. Eighty-nine percent (144/161 eyelids) achieved the target result of an MRD1 of 4 +/- 1 mm after one procedure. Fifteen eyelids (9% of operated eyelids) required a second procedure, and in this group, 2 (13% of the reoperated eyelids) underwent a third procedure. Although bilateral cases were more likely to achieve symmetry (p = 0.0071), 90% of either unilateral or bilateral cases achieved a postoperative MRD1 of 4 +/- 1 mm. Both mild (MRD1 of 5-7 mm) and severe (MRD1 > 7 mm) cases of eyelid retraction achieved similar operative outcomes. In the first 6 months after surgery, complications included undercorrection (8 eyelids), overcorrection (2 eyelids), and pyogenic granuloma (2 eyelids). None had a flattened upper eyelid contour. Late recurrence of retraction was seen in 9 eyelids. Mean operative time was 16 minutes per eyelid. CONCLUSIONS: This technique of lowering the retracted upper eyelid is effective even in severe cases of eyelid retraction. Minimal complications were encountered, and upper eyelid contours were well preserved. The use of hot-wire cautery dissection proved useful in shortening operative time. PMID- 17117097 TI - Acquired ptosis secondary to vernal conjunctivitis in young adults. AB - PURPOSE: To establish a relation between prolonged severe vernal conjunctivitis and upper eyelid ptosis. METHODS: The study consisted of 12 patients between the ages of 19 and 32 years with acquired ptosis who presented in our clinic between September 2001 and February 2005. Potential factors responsible for acquired ptosis were investigated in all patients, with specific attention to the history and severity of vernal conjunctivitis. RESULTS: We found vernal conjunctivitis to be the identifiable cause in 8 men and 4 women with acquired ptosis. There was neither contact lens usage nor trauma or ocular surgery history in their medical records. The blepharoptosis was caused by levator disinsertion and recession of the aponeurosis. The pathology improved in each case after reattachment of the aponeurosis to the superior tarsal border. CONCLUSIONS: These findings suggest that prolonged severe vernal conjunctivitis may induce a lower position of the upper eyelid and eventually lead to ptosis through levator disinsertion that is similar to involutional ptosis. We believe that chronic inflammation of the upper eyelid with giant papillary conjunctivitis and persistent rubbing of the eyelids may be responsible for the development of this pathology. PMID- 17117098 TI - Frontalis muscle flap advancement with a pulley in the levator aponeurosis in patients with complete ptosis and deep-set eyes. AB - PURPOSE: To offer an alternative to frontalis sling techniques to achieve a more horizontal traction vector force with the frontalis muscle flap. The proposed technique avoids malpositions such as pulling the upper eyelid in an anterior plane during maximum opening and ptosis of the eyelashes. METHODS: The technique, performed in 5 patients with complete unilateral ptosis and an absence of levator function, consisted of advancing a flap of frontalis muscle and creating a pulley with the aponeurosis of the levator muscle. Mean follow-up was 12 months. RESULTS: Ptosis was corrected in all 5 patients, with good aesthetic outcome. There were no cases of anterior eyelid advancement, entropion, or eyelash ptosis. The only complication was 1 case of lagophthalmos with corneal erosion. CONCLUSIONS: In this preliminary series, the technique was safe and effective for correcting severe blepharoptosis, with good aesthetic and functional results even in patients with deep-set eyes and without the eyelid malpositions that commonly occur in frontalis sling techniques. PMID- 17117099 TI - Treatment of trichiasis using an 810-nm diode laser: an efficacy study. AB - PURPOSE: To determine the efficacy of an 810-nm diode laser in the treatment of trichiasis. METHODS: Noncomparative interventional prospective study of 87 patients, providing 153 eyelids with darkly pigmented trichiatic eyelashes, treated with the 810-nm diode laser. The number of trichiatic eyelashes was counted before and after treatment. Patients were followed for 6 months with monthly examinations. RESULTS: Immediately after laser treatment, the average number of trichiatic eyelashes per patient was reduced from 3.50 (standard deviation, 4.20) to 0.40 (standard deviation, 1.0). Forty-one patients were followed for 3 or more months (minimum of 90 days; maximum, 443 days; standard deviation, 77.23 days). Among these patients, the average number of eyelashes per patient was reduced from 3.58 (standard deviation, 4.15) to 0.73 (standard deviation, 1.60). The paired t tests demonstrated a statistically significant difference between numbers of eyelashes before and after laser treatment irrespective of gender, age, or location of eyelashes. CONCLUSIONS: The 810-nm diode laser is an effective tool in treating trichiasis. PMID- 17117100 TI - Efficacy of botulinum toxin type a after topical anesthesia. AB - PURPOSE: To determine whether the use of topical anesthesia has an impact on botulinum toxin type A (BTX-A) efficacy. METHODS: Forty patients (20 receiving BTX-A for facial cosmetic rhytid reduction and 20 for benign essential blepharospasm) were evaluated in a double-blind, randomized, triple-crossover study at 2.5- to 4.5-month intervals. The discomfort and efficacy of BTX-A injections after betacaine application to half the face (random assignment) were compared against the discomfort and efficacy of a placebo ointment on the other half of the face. This was followed by cryoanalgesia to the entire face. RESULTS: Patients ranged from 27 to 81 years of age (mean, 53 years), and 34 were female. Of the 120 total injection comparisons, a better BTX-A effect on one side of the face was reliably identified by 80% and 77% of blepharospasm and cosmetic patients, respectively, with the placebo-treated side providing better BTX-A effect approximately 90% of the time (p < 0.001). Patients reported a more painful side during injection in just 18 of the 120 trials, and only 1 of 40 patients believed the administration of analgesia was worth the trouble. CONCLUSIONS: Pretreatment with topical betacaine followed by skin cooling seems to have a deleterious impact on BTX-A effect without a significantly beneficial patient-perceived reduction in injection discomfort. PMID- 17117101 TI - Duration of botulinum toxin effect in the treatment of crocodile tears. AB - PURPOSE: To provide clinical evidence of the duration of botulinum toxin type A (BTX-A) effect when applied in the palpebral lobe of the lacrimal gland in patients with gustatory epiphora. METHODS: Prospective, nonrandomized, nonblinded study. Patients with history of gustatory epiphora were included. A Schirmer test was performed to quantify tearing induced by chewing. Clinical examination included visual acuity, tear-duct syringing, slit lamp examination, corneal staining, and eyelid malpositions. A questionnaire was completed by each patient to asses the severity of hyperlacrimation. A single dose of 2.5 units of BTX-A was injected directly into the lacrimal gland palpebral lobe. Patients were evaluated before and at 1, 4, 12, and 24 weeks after injection. The same person performed the examination and the BTX-A injection. Descriptive statistics, using repeated measures and a paired t test, were used for statistical analysis. RESULTS: Fifteen patients were included. Mean age was 63 years. Before BTX-A injection, mean Schirmer test values were 5.47 mm in the unaffected eyes (NAE) and 12.07 mm in the affected eyes (AE). When comparing Schirmer test values in the AE before and after BTX-A injection, there were statistically significant differences (p < 0.05). Only 2 patients developed mild transitory ptosis. No other complications were noted. CONCLUSIONS: The effect of 2.5 units of BTX-A injected into the lacrimal gland lasted 6 months, a duration similar to that reported for other application sites. PMID- 17117102 TI - Prominent premalar and cheek swelling: a sign of thyroid-associated orbitopathy. AB - PURPOSE: To describe prominent premalar and cheek swelling as a previously undescribed clinical feature of thyroid-associated orbitopathy (TAO). DESIGN: Retrospective interventional case series. METHODS: A retrospective case review of patients with prominent premalar and/or cheek swelling and TAO was undertaken. All patients who presented from March 2002 to February 2005 with complaints of premalar and/or cheek swelling in TAO were analyzed. RESULTS: Six female patients between the ages of 28 and 66 years (average, 44.2 years) who had complaints of prominent premalar and/or cheek swelling and TAO were included in the study, among 326 new patients with TAO (incidence: 1.84%). Five of the 6 patients had Graves hyperthyroidism. One initially had Hashimoto thyroiditis, which converted to Graves hyperthyroidism. In all cases, TAO preceded the thyroid disease or developed simultaneously (average time from TAO to thyroid disease was 3.0 months). No patient had received corticosteroids before premalar and/or cheek swelling. The premalar and/or cheek swelling was bilateral in all cases, but 2 of 6 were asymmetric. No diurnal fluctuation or tenderness in premalar and/or cheek swelling was noted. All improved incompletely over several months. Brow, eyelid swelling, and orbital-fat hypertrophy on radiologic examination was coincidentally noted in 5 of 6 cases. Pretibial myxedema was noted in one case. One case was associated with another autoimmune disease (rheumatoid arthritis). CONCLUSIONS: Prominent premalar and cheek swelling should be considered among the clinical features of TAO. The true incidence will become apparent as we question and examine patients more carefully regarding this entity and as we review premorbid photographs. PMID- 17117103 TI - Implant infection in porous orbital implants. AB - PURPOSE: To analyze implant infection in patients with porous orbital implants. METHODS: A retrospective analysis of 212 patients with one of five types of porous orbital implants (bone-derived hydroxyapatite [HA], coralline HA, synthetic HA, porous polyethylene, and aluminium oxide) was conducted. Reasons for surgery, type of surgery, type of implant, peg system used, time of pegging, problems before and after pegging, treatment, and follow-up duration were recorded for all patients, along with additional data including time of onset of infection, microorganism cultured, antibiotics used, patient response to antibiotic therapy, additional interventions, and final status for patients with infection. RESULTS: Of the 212 patients with porous orbital implants, 116 (54.72%) were pegged. Implant infection was observed in 11 of 116 patients (9.48%) with pegs, whereas 0% of unpegged implants was infected (p = 0.001). The interval between pegging and the onset of infection was 3 to 83 months (average, 36.27 +/- 29.12 months). Implant exposure was noted in 5 of the 11 patients with infection. Symptoms resolved completely with antibiotic treatment in 7 patients. One patient required implant removal as the result of frequent exacerbations. The remaining 3 patients presented with hemorrhagic, purulent discharge and/or pyogenic granuloma on their last visits after being free of symptoms for 5 to 6 months. CONCLUSIONS: Implant infection is a serious problem that requires additional patient visits, intensive antibiotic therapy, surgery, or some combination of these. Existence of a peg system appears to play a role in implant infection. Infection may develop as late as 6 to 7 years after pegging, and the patient should be cautioned about potential late-onset problems. It is possible to control the infection with appropriate antibiotic therapy; removal should be reserved for refractory cases. PMID- 17117104 TI - Pyogenic granuloma as a presenting sign of hydroxyapatite orbital implant exposure: a clinicopathologic study. AB - PURPOSE: We report 5 unusual cases of exposed hydroxyapatite orbital implants that presented as pyogenic granulomas. We propose pathogenesis and histopathologic correlations. METHODS: A clinicopathologic study of 5 patients with hydroxyapatite implants who presented with pyogenic granuloma. RESULTS: Pyogenic granulomas were detected 1.5 to 30 months after implantation in 5 patients. The lesions were multiple but were not related to the wound margin at the exposed area and were not covered by the surface epithelium in most instances. Exposure defects were detected in all patients at the time of lesion excision. The mean exposure size in the greatest dimension was 16 mm (range, 9-20 mm). Three patients were treated successfully with simple excision of the granulomas, burring down of the anterior surface of the implants, and direct repair of the exposure defects. Explantation of the implant was performed in 2 cases. Histopathologic examination revealed chronic inflammation and microabscess formation in the explanted implants. CONCLUSIONS: Five patients with pyogenic granulomas were found to have hydroxyapatite exposure. Pyogenic granuloma should not be considered a benign lesion on a hydroxyapatite orbital implant, especially in recurrent cases. Ophthalmologists must be aware of the possibility of conjunctival dehiscence with hydroxyapatite-implant exposure beneath the lesion. PMID- 17117105 TI - Blinding orbital cellulitis: a complication of strabismus surgery. AB - A 56-year-old healthy man underwent left medial rectus recession and lateral rectus resection for esotropia. The next day he developed severe left periocular pain with decreased vision, an afferent pupillary defect, periorbital edema, limited ocular motility, and proptosis. Computed tomography showed fat stranding and less than 90 degrees of posterior globe tenting. Despite intravenous antibiotics to treat orbital cellulitis, and a lateral canthotomy and cantholysis to decompress the orbit, visual acuity worsened to no light perception. The patient underwent emergent orbital decompression including release of the superior and inferior septum and outfracturing of the orbital floor and medial wall; however, there was no recovery of vision. Blinding orbital cellulitis is a rare complication after strabismus surgery. Despite poor prognosis, prompt diagnosis and aggressive treatment may maximize visual potential. PMID- 17117106 TI - Intraorbital heroin injection resulting in orbital cellulitis and superior ophthalmic vein thrombosis. AB - A 47-year-old man with decreased vision, ophthalmoplegia, proptosis, and chemosis of his right eye admitted to injecting heroin directly into his orbit. He was placed on intravenous antibiotics for orbital cellulitis, and computed tomography and magnetic resonance imaging were performed. Superior ophthalmic vein thrombosis (SOVT) was noted on magnetic resonance imaging. The patient responded well to intravenous antibiotics, and his symptoms resolved with minimal deficits. Steroids and anticoagulants were not administered. We review the pathogenesis of septic SOVT and briefly discuss the role of anticoagulants and steroids in this setting. PMID- 17117107 TI - Fornix reconstruction with conjunctival inclusion cyst marsupialization in Stevens-Johnson syndrome. AB - A 17-year-old boy presented with multiple, progressively enlarging cystic masses located at the junction of the lower eyelids and anterior orbit, 1 year after recovery from Stevens-Johnson syndrome. Bilaterally, symblepharon obliterated the inferior fornix and restricted supraduction. All but 1 cyst (which was sent for microscopic evaluation) was marsupialized. Cut edges of the cysts were sutured to adjacent conjunctiva and the epithelial walls were used as fornix lining. Histologically, the wall of the excised cyst consisted of conjunctiva. One year after surgery, the inferior fornices remained substantially deepened with improved ocular motility. This case demonstrated that conjunctival inclusion cysts occurring in the aftermath of Stevens-Johnson syndrome can be effectively managed with marsupialization. The epithelial lining can then be used in fornix reconstruction. PMID- 17117108 TI - Eyelid switch flap technique for the management of congenital anophthalmos associated with contracted socket. AB - Congenital anophthalmos with contracted socket can occasionally present in the second or third decade for cosmetic correction of the deformity. Placement of ocular prosthesis is not possible due to reduced dimensions of the palpebral aperture and contracture of the bony socket. Horizontal widening of the palpebral aperture using an eyelid switch flap technique followed by placement of ocular prosthesis anterior to the hypoplastic orbit achieves good cosmetic correction of this deformity. PMID- 17117109 TI - Eyelid retraction and incomplete eyelid closure secondary to human immunodeficiency virus-associated muscle wasting. AB - Human immunodeficiency virus (HIV)-associated weight loss remains a significant problem, even in the era of highly active antiretroviral therapy. This interventional case report describes eyelid retraction and poor eyelid closure caused by orbicularis atrophy in the setting of HIV-associated muscle wasting. A 65-year-old HIV-infected man sought treatment for chronic ocular irritation. On examination, he was thin with marked temporal wasting. Corneal epithelial defects were present bilaterally. There was 2 mm of superior scleral show in the right eye and trace inferior scleral show bilaterally. With attempted closure, lagophthalmos approached 1 cm in the right eye and was 3 mm in the left eye. The remainder of the examination was unremarkable. Gold weight placement achieved symptomatic improvement with adequate eyelid closure. Biopsy demonstrated fibrous tissue with an absence of identifiable muscle fibers. In the setting of HIV associated muscle wasting, orbicularis oculi muscle atrophy may result in eyelid retraction, lagophthalmos, and ocular surface disease. PMID- 17117110 TI - Effect of 7.0 Tesla MRI on upper eyelid implants. AB - To ensure the MRI compatibility of various eyelid implants in high-field MRI, 3 eyelid weights made of pure gold (99.99%), pure platinum (99.95%), and a platinum (97%)/iridium (3%) alloy were examined in vitro. Temperature changes, position changes, and imaging artifacts of the different implants were determined in a small-bore 7.0 Tesla MRI system. The 7.0 Tesla MRI system demonstrated that none of the eyelid implants carried a risk of heating or dislocation; therefore, these implants are MRI compatible up to a magnetic field strength of 7.0 Tesla. PMID- 17117111 TI - MR findings in a patient with isolated intrinsic optic nerve lymphoma. AB - We present a case of unilateral isolated intrinsic optic nerve small B-cell lymphoma confirmed by optic nerve biopsy in a patient with lymphoplasmacytoid lymphoma. Brain and orbital MRI with gadolinium revealed fusiform enlargement of the optic nerve from the level of the optic nerve head to the prechiasmatic region, with associated homogeneous enhancement of the nerve itself. PMID- 17117112 TI - Orbital chondroma. AB - A 9-year-old boy was referred for increasing ptosis in the left eye. CT and MRI revealed a well-circumscribed mass in the superonasal orbit. An anterior orbitotomy was performed and the mass was excised in toto. Pathologic evaluation revealed a pure chondroma. Chondroma is an uncommon tumor that may rarely present as an orbital mass. PMID- 17117113 TI - Orbital amyloidosis-induced compressive optic neuropathy accompanied by characteristic eyelid pigmentation. AB - The deposition of amyloid protein in orbital tissue is called orbital amyloidosis. Orbital amyloidosis is a rare condition that usually affects older patients. Although the disease is slowly progressive, it rarely involves the optic nerve or threatens vision. We report a case of orbital amyloidosis that initially appeared as impressive eyelid pigmentation and blepharoptosis and then progressed to systemic amyloidosis. The orbital lesion induced compressive optic neuropathy. Surgical management enabled partial but not full recovery of vision. PMID- 17117114 TI - Spontaneous orbital and periocular hemorrhage in a patient with epithelioid hemangioma. AB - Epithelioid hemangioma, or angiolymphoid hyperplasia with eosinophilia, is a benign vascular lesion that infrequently involves the orbit. We report a case of spontaneous orbital and periocular hemorrhage in a patient with epithelioid hemangioma of the orbit. Ophthalmic examination was accompanied by CT and MRI, followed by histopathologic diagnosis. The patient presented with headache, eyelid bruising, and double vision. Ophthalmic examination showed ecchymosis, proptosis, and limited extraocular movements of the affected eye. Imaging studies showed a cystic right orbital mass. Histologic examination showed plump endothelial cells accompanied by lymphocytes, plasma cells, and eosinophils. A diagnosis of epithelioid hemangioma was rendered. To our knowledge, this is the first reported case of spontaneous orbital and periocular hemorrhage associated with epithelioid hemangioma. Epithelioid hemangioma should be considered in the differential diagnosis for patients presenting with spontaneous orbital or periocular hemorrhage. PMID- 17117115 TI - Unusual presentation of gastric adenocarcinoma metastatic to the orbit. AB - A 69-year-old woman with no history of malignant disease presented with complaints of ptosis, diplopia, and left upper eyelid fullness. Computed tomography showed soft tissue infiltration of the left superior orbit. Biopsy was performed through an anterior orbitotomy. Histopathology revealed a mucinous adenocarcinoma. Subsequent systemic evaluation included esophagogastroduodenoscopy, which revealed a primary gastric malignancy. PMID- 17117116 TI - Bilateral periorbital ecthyma gangrenosum. AB - We describe a case of bilateral periorbital ecthyma gangrenosum in a diabetic patient with renal failure. Ecthyma gangrenosum is a cutaneous manifestation of Pseudomonas sepsis. We briefly review the pathogenesis of ecthyma gangrenosum and discuss previous reports of periocular involvement. In our patient, conservative measures and supportive care of the periorbital tissue resulted in a good outcome. PMID- 17117117 TI - Nodular fasciitis presenting as a large mass in the upper eyelid. AB - A 3-year-old boy was examined for a large nodular fasciitis in the upper eyelid. The lesion was confined to the preseptal plane of the upper eyelid, provoking mechanical ptosis. Excision by an eyelid crease approach was sufficient to restore normal eyelid position. PMID- 17117118 TI - Kimura disease of the eyelid: a clinicopathologic study with electron microscopic observations. AB - A 70-year-old white man presented with a solitary eyelid nodule of 6 months' duration. Because lymphoma could not be excluded on the initial biopsy, the mass was completely excised. Histologic examination confirmed the diagnosis of Kimura disease, an uncommon chronic inflammatory condition. Peripheral eosinophilia was present. The patient did not have any other lesions of Kimura disease on follow up examination. The clinical, radiologic, histologic, and electron microscopic findings of Kimura disease are discussed. To our knowledge, solitary involvement of the eyelid has not been reported previously. Kimura disease should be included in the differential diagnosis of an eyelid mass lesion. PMID- 17117119 TI - Aggravation of eyelid and conjunctival malignancies following photodynamic therapy in DeSanctis-Cacchione syndrome. AB - A 5-year-old girl with DeSanctis-Cacchione syndrome (a severe variant of xeroderma pigmentosum) was referred for evaluation of multiple eyelid and bulbar conjunctival squamous cell carcinomas. Examination evidenced multiple vegetating lesions on the eyelid, bulbar conjunctiva, and cornea of both eyes. As the lesions were considered not to be manageable by surgical excision and would have required exenteration, photodynamic therapy was performed on the patient's left eye. Three months after photodynamic therapy, the patient presented with a dramatic increase in the extension of the tumors. Since xeroderma pigmentosum is a DNA repair disorder, the radiation involved in photodynamic therapy probably played an iatrogenic role in the evolution of the case. We believe that photodynamic therapy may be harmful to patients with DNA repair disorders. PMID- 17117120 TI - A case of sneezing-related orbital emphysema treated by aspiration-decompression in the office. PMID- 17117122 TI - Reply re: "Lateral tarsal strip and Quickert sutures for lower eyelid entropion". PMID- 17117123 TI - A doctor's dilemma. PMID- 17117124 TI - Use of lone star elastic stay hooks in orbital surgery. PMID- 17117126 TI - Gender influences in vivo human responses to endotoxin. AB - Gender appears to influence systemic and organ-specific inflammatory sequelae of ischemia-reperfusion and infectious challenge in many animal models. Despite the protection provided by female gender, androgen blockade, and/or estrogen administration in such experimental studies, many questions remain regarding the influence of gender dimorphism upon human responses to injury. We hypothesized that the administration of low-dose lipopolysaccharide (LPS) to otherwise healthy, young adults would provide insights regarding the influence of gender upon physiological and innate immune system responses to a prototypic inflammatory stimulus. To this end, 72 adult subjects (48 men, aged 29 +/- 1.0 years; 24 women, aged 26 +/- 1.0 years) were prospectively evaluated before and after the i.v. administration of LPS (2 ng/kg). All subjects developed symptoms within 1.0 to 1.5 h after LPS, and the men exhibited a greater increase in core temperature (2.1 +/- 0.1 degrees C) compared with the women (1.4 +/- 0.1 degrees C) (P < 0.001). In addition, the men exhibited a greater maximum decrease in mean arterial pressure (-13.0 +/- 1.3 mmHg) compared with the women (-8 +/- 1.3 mmHg) (P < 0.02). The changes in temperature and mean arterial pressure occurred without detectable differences between the male and female cohort responses of circulating white blood cell count and cortisol or cytokine levels. These results suggest that soluble inflammatory mediators generated by in vivo endotoxin activation of the innate immune system are insufficient to explain the resultant gender-specific phenotypic differences observed in young, adult humans. PMID- 17117127 TI - The occurrence of severe sepsis and septic shock are related to distinct patterns of cytokine gene expression. AB - Patient response to acute bacterial infection is highly variable. Differing outcomes in this setting may be related to variations in the immune response to an infectious insult. Using quantitative real-time polymerase chain reaction, we quantified gene expression of the tumor necrosis factor alpha(TNFalpha), interferon gamma (IFNgamma), and interleukin 10 (IL10), IL12p35, and IL4 genes in 3 patient groups. These groups consisted of an intensive care unit (ICU) cohort who presented with severe sepsis or septic shock, a group of noncritically ill ward patients with documented Gram-negative bacteremia, and a group of healthy controls. Greater interleukin 10 messenger RNA (mRNA) levels were detected in the ICU group in comparison with both the bacteremic and control groups (P < 0.0001). More TNF-alpha mRNA was detected in the ICU group when compared with the control group (P < 0.0001). However, TNF-alpha mRNA was most abundant in the bacteremic group (P = 0.0007). Lesser IFN-gamma mRNA levels were detected in the ICU group when compared with both the bacteremic and control groups (P < 0.0003). Cytokine mRNA levels were not associated with the occurrence of shock upon admission to ICU. On the seventh day of ICU stay, the presence of shock was associated with lesser IFN-gamma mRNA (P = 0.0004) and lesser TNF-alpha mRNA (P = 0.001). Survivors had greater TNF-alpha mRNA copy numbers on day 7 of ICU stay than nonsurvivors (P = 0.002). We conclude that a proinflammatory response is the appropriate response in the setting of infection and is associated with lesser requirements for inotropes and lesser mortality. Quantitative real-time polymerase chain reaction can be used to predict infection outcome in clinically relevant situations where enzyme-linked immunosorbent assay testing has proved disappointing. PMID- 17117128 TI - A modified goal-directed protocol improves clinical outcomes in intensive care unit patients with septic shock: a randomized controlled trial. AB - We evaluated whether a goal-directed protocol, without measurement of central venous oxygen saturation, would improve survival in medical intensive care unit (ICU) patients with septic shock. This is a prospective, controlled study in a 24 bed medical ICU at a tertiary care hospital. From a total of 241 consecutive patients with septic shock, 224 were randomly assigned to receive therapy with or without a written protocol using central venous pressure, mean arterial pressure, and urine output as therapeutic goals. Baseline characteristics were similar between groups. Implementation of goal-directed therapy caused a more rapid reversal of persistent shock (47 +/- 22.8 vs. 65.4 +/- 32.1 h, P = 0.006) and decreases of ICU (50% vs. 67.2%, P = 0.009) and in-hospital (53.7% vs. 71.6%, P = 0.006) mortality rates compared with non-goal-directed therapy. Patients receiving goal-directed therapy also had less risk for developing central nervous system or renal failure than patients without. Patients with goal-directed therapy received more fluid during the period of persistent shock (136.2 +/- 119 vs. 88.6 +/- 57.7 mL h, P = 0.034) and less delay in vasopressor administration (78 +/- 22.2 vs. 104.4 +/- 29 min, P = 0.001) than patients with non-goal therapy. Implementation of a goal-directed protocol improves survival and clinical outcomes in ICU patients with septic shock. These benefits may arise from adequate fluid resuscitation, earlier vasopressor administration, rapid shock reversal, and protection of major organ function. With central venous oxygen saturation measurement to detect tissue perfusion, the clinical outcomes may be further improved. PMID- 17117129 TI - Aberrant regulation of polymorphonuclear phagocyte responsiveness in multitrauma patients. AB - A systemic inflammatory response often follows severe trauma. Priming (preactivation) of polymorphonuclear phagocytes (PMNs) is an essential first step in the processes that lead to damage caused by the systemic activation of innate immune response. Until recently, priming could only accurately be measured by functional assays, which require isolation of cells, thereby potentially inducing artificial activation. The aim of this study was to identify primed PMNs in response to trauma by using a whole blood analysis with a broad detection range. Twenty-two trauma patients were analyzed for PMN priming with novel developed antibodies recognizing priming epitopes by flow cytometric analysis. Expression of priming epitopes on PMNs was analyzed with respect to time, injury, and disease severity. Expression of priming epitopes in the circulation was compared with expression profiles of PMNs obtained from lung fluid. Fourteen healthy volunteers served as controls. Expression of priming epitopes on peripheral blood PMNs of injured patients was similar, as found in healthy controls, whereas highly primed cells were found in the lung fluid of injured patients (increase of >50 times as compared with peripheral blood cells). In fact, the responsiveness of PMNs toward the bacteria-derived stimulus N-formyl-methionyl-leucyl phenylalanine was markedly decreased in trauma patients. Lack of expression of priming epitopes and the unresponsiveness to N-formyl-methionyl-leucyl phenylalanine demonstrates the presence of partially refractory cells in the circulation of trauma patients. An increased expression of epitopes found on pulmonary PMNs suggests that optimal (pre)activation of these cells only occurs in the tissues. PMID- 17117130 TI - Pneumonia after cecal ligation and puncture: a clinically relevant "two-hit" model of sepsis. AB - Sepsis continues to be the primary cause of death among patients in surgical intensive care units. In many cases, death does not result from the initial septic event but rather from subsequent nosocomial infection with pneumonia being the most common etiology. In addition, most deaths in patients with sepsis occur after the first 72 h. By contrast, in most animal models of sepsis, most deaths occur within the first 72 h. The purpose of this study was to develop a clinically relevant "two-hit" model of sepsis that would reflect delayed mortality because of secondary nosocomial infection. The well-accepted and widely used cecal ligation and puncture (CLP) model was used as the "first hit". Pseudomonas aeruginosa or Streptococcus pneumoniae was used to induce pneumonia in mice 72 h after CLP as a "second hit." In this study, mortality in mice undergoing CLP followed by pneumonia was significantly higher than in mice receiving pneumonia or CLP alone. S. pneumoniae pneumonia after CLP resulted in a 95% mortality compared with a 20% mortality for pneumonia alone, P < 0.0001. Similarly, mortality of P. aeruginosa pneumonia after CLP (85%) was significantly higher than P. aeruginosa alone (20%), P < 0.0001. Mice undergoing CLP followed by P. aeruginosa pneumonia also had significantly higher levels of B- and T-cell apoptotic death. Finally, mice undergoing CLP followed by P. aeruginosa or S. pneumoniae pneumonia had significantly decreased concentrations of proinflammatory mediators monocyte chemoattractant protein-1 and interleukin (IL) 6 compared with mice undergoing CLP or pneumonia alone. In conclusion, a primary sublethal infection impairs the immune system thus rendering the host more susceptible to secondary infection and death. Double injury, that is, CLP followed by pneumonia, provides a useful tool in the study of sepsis, creating a prolonged period of infection as opposed to CLP alone. The extended duration of infection may lead to a better understanding of the mechanism of the immune dysregulation seen in clinical sepsis and therefore provides for evaluation of potential therapies that target specific stages of the immune response. PMID- 17117131 TI - Sex dimorphisms in activated mesenchymal stem cell function. AB - The plasticity of bone marrow-derived stem cells (BMSCs) has resulted in positive remodeling and the regeneration of viable tissues. However, BMSC release of growth factors, which limit apoptosis and inflammation, may play an important role in conferring organ protection. Recent studies also indicate that those patients with higher circulating BMSC counts may be more resistant to septic and traumatic insults. There are clear sex differences in response to such insults. Within the population of BMSC, mesenchymal stem cells (MSCs) may have clinical advantages. Therefore, we hypothesize that sex differences in the MSC paracrine response to acute injury exist. Mesenchymal stem cells were obtained from male and female mice. One million MSCs per well (triplicate wells per group) were stressed by hypoxia and increasing doses of endotoxin (lipopolysaccharide [LPS]) and hydrogen peroxide. Mesenchymal stem cell activation was determined by measuring vascular endothelial growth factor (VEGF) and tumor necrosis factor alpha production by enzyme-linked immunosorbent assay. Differences were considered significant if P < 0.05. RESULTS: Lipopolysaccharide resulted in significant activation of both male and female MSCs. However, LPS provoked significantly more VEGF production in female MSCs versus male MSCs at all LPS doses. Hypoxia of 1 h and hydrogen pyroxide exposure also caused significantly more VEGF production in female MSCs versus male MSCs. Female MSCs expressed significantly less tumor necrosis factor alpha than male MSCs after acute LPS and hypoxia. CONCLUSION: This study constitutes the first demonstration that sex differences exist in activated MSC function. Sex differences in progenitor cell function may have important implications in understanding the observed sex differences in the host's response to injury. PMID- 17117132 TI - High passage number of stem cells adversely affects stem cell activation and myocardial protection. AB - Progenitor cell plasticity enhances positive remodeling of damaged tissue. We and others have previously shown that progenitor cells may limit apoptosis and modulate inflammation in part by the production of growth factors. However, recent studies suggest that progenitor cells senesce and lose their differentiation potential with increasing time in culture and passage. We hypothesize that murine bone marrow mesenchymal stem cells (MSCs) are cardioprotective against ischemia/reperfusion injury in the isolated perfused rat heart, and that passage number has an adverse effect on MSC activation and cardioprotection. Adult male and female Sprague-Dawley rat hearts were isolated, perfused via Langendorff model, and subjected to ischemia/reperfusion. Mouse MSCs were harvested, cultured, suspended in perfusate, and infused before global index ischemia. Hearts were assigned to controls or infusion with passage 3, 5, or 10 MSCs. In addition, MSCs in culture were stressed by hypoxia and increasing doses of endotoxin (lipopolysaccharide). Mesenchymal stem cell activation was determined by measuring vascular endothelial growth factor production with enzyme linked immunosorbent assay. All data are reported as mean +/- SEM and were analyzed with 2-way analysis of variance. Differences are considered significant if P < 0.05. Passage 3 murine MSC infusion in hearts before ischemia reduced the depression of left ventricular developed pressure, attenuated the increase of end diastolic pressure, and reduced the depression of +dP/dT and -dP/dT. However, the MSC protective effect disappeared in hearts infused with passage 5 and passage 10 MSCs. Although hypoxia and lipopolysaccharide resulted in significant activation of MSCs, passage 3 MSCs demonstrated significantly greater vascular endothelial growth factor release than passage 5 and 10 MSCs. Acute murine MSC infusion confers protection in isolated rat hearts. However, high passage number has an adverse effect on MSC activation and protection. This portends limited ex vivo expansion before possible therapeutic use. PMID- 17117133 TI - Synergistic effects of recombinant human soluble thrombomodulin and fluid-volume resuscitation in a rat lethal crush injury model. AB - Severe crush injury results in a high mortality rate because of acute circulatory failure and hyperkalemia. The purpose of this study was to evaluate whether administration of prophylactic-recombinant human soluble thrombomodulin (rhsTM) and/or fluid-volume resuscitation before reperfusion attenuates severe crush injury in rats. Both hindlimbs of anesthetized rats were compressed for 6 h under blocks weighing 3.5 kg each, followed by 3 h of reperfusion. In the first group, fluid resuscitation with normal saline (1 mL/kg/h) was performed throughout the experiment. In the second group, volume resuscitation treatment with normal saline (10 mL/kg/h) was initiated 60 min before the end of the crush period and was continued until the end of the experiment. In the third group, normal saline resuscitation treatment plus rhsTM (3 mg/kg) was performed. In the fourth group, volume resuscitation treatment plus rhsTM was performed. Blood samples were collected 6 h after the end of the crush period. Complete blood count and platelets were measured. In addition, serum lactate, base deficit, serum potassium, creatine phosphokinase, blood urea nitrogen, creatinine, myoglobin, and some cytokines were evaluated. In another experiment, survival of each group was monitored for 72 h after the end of the crush period. Combined administration of rhsTM and volume resuscitation significantly decreased hemoconcentration and hyperkalemia. The serum interleukin-6 level and mortality were also significantly improved in the combination group compared with those in the other groups. We conclude that prophylactic combination of rhsTM administration and volume resuscitation may be an effective therapy for severe crush injury. PMID- 17117134 TI - Chemically modified tetracycline (COL-3) improves survival if given 12 but not 24 hours after cecal ligation and puncture. AB - Sepsis can result in excessive and maladaptive inflammation that is responsible for more than 215,00 deaths per year in the United State alone. Current strategies for reducing the morbidity and mortality associated with sepsis rely on treatment of the syndrome rather than prophylaxis. We have been investigating a modified tetracycline, COL-3, which can be given prophylactically to patients at high risk for developing sepsis. Our group has shown that COL-3 is very effect at preventing the sequelae of sepsis if given before or immediately after injury in both rat and porcine sepsis models. In this study, we wanted to determine the "treatment window" for COL-3 after injury at which it remains protective. Sepsis was induced by cecal ligation and puncture (CLP). Rats were anesthetized and placed into five groups: CLP (n = 20) = CLP without COL-3, sham (n = 5) = surgery without CLP or COL-3, COL3@6h (n = 10) = COL-3 given by gavage 6 h after CLP, COL3@12h (n = 10) = COL-3 given by gavage 12 h after CLP, and COL3@24h (n = 20) = COL-3 given by gavage 24 h after CLP. COL-3 that was given at 6 and 12 h after CLP significantly improved survival as compared with the CLP and the CLP@24h groups. Improved survival was associated with a significant improvement in lung pathology assessed morphologically. These data suggest that COL-3 can be given up to 12 h after trauma and remain effective. PMID- 17117135 TI - The role of initial trauma in the host's response to injury and hemorrhage: insights from a correlation of mathematical simulations and hepatic transcriptomic analysis. AB - Trauma and hemorrhagic shock (HS) elicit severe physiological disturbances that predispose the victims to subsequent organ dysfunction and death. The general lack of effective therapeutic options for these patients is mainly due to the complex interplay of interacting inflammatory and physiological elements working at multiple levels. Systems biology has emerged as a new paradigm that allows the study of large portions of physiological networks simultaneously. Seeking a better understanding of the interplay among known inflammatory pathways, we constructed a mathematical model encompassing the dynamics of the acute inflammatory response that incorporates the intertwined effects of inflammation and global tissue damage. The model was calibrated using data from C57Bl/6 mice subjected to endotoxemia, sham operation (i.e., surgical trauma induced by cannulation [ST]) or ST + HS+ resuscitation (ST-HS-R). An in silico simulation, made at whole-organism level, suggested that similar pathways of different magnitudes were operant as the degree of total body damage increased. We sought to validate this hypothesis by subjecting mice to HS and comparing the models predictions to circulating markers of inflammation and tissue injury as well as the global transcriptomic response of the liver. C57Bl/6 mice were subjected to ST or ST-HS (without resuscitation). Liver gene expression was assessed using an Affymetrix DNA microarray (GeneChip Mouse Expression Set 430A, Affymetrix, Santa Clara, CA), which contains 22,621 probe sets and effectively interrogates 12,341 mouse genes. The microarray data sets were subjected to hierarchical clustering and pathway analysis. In agreement with model predictions, circulating levels of inflammation/tissue injury markers and the microarray analysis both demonstrated that ST alone accounts for a substantial proportion of the observed phenotypic and genetic/molecular changes versus untreated animals. The addition of HS further increased the magnitude of gene expression, but relatively few additional genes were recruited. Mathematical simulations and DNA microarrays, both systems biology tools, may provide valuable insight into the complex global physiological interactions that occur in response to trauma and hemorrhagic shock. PMID- 17117136 TI - Platonin, a cyanine photosensitizing dye, is effective for attenuation of heatstroke in rats. AB - The present study was performed to assess the prophylactic effect of platonin, a cyanine photosensitizing dye and an inhibitor of proinflammatory cytokines, in an animal model of heatstroke. Anesthetized rats were immediately divided into 2 major groups after the start of heat stress and administered either isotonic sodium chloride solution (dose, 1 mL/kg of body weight i.v.) or platonin (dose, 12.5-50 microg/mL per kilogram of body weight i.v.). They were exposed to ambient temperature of 43 degrees C to induce heatstroke. Another group of rats were exposed to room temperature (26 degrees C) and used as normothermic controls. Their physiological and biochemical parameters were continuously monitored. When the isotonic sodium chloride solution-pretreated rats underwent heat stress, their survival time values were found to be from 20 to 24 min. Pretreatment with intravenous doses of platonin (12.5-50 microg/mL per kilogram of body weight) immediately after the start of heat exposure significantly improved survival time during heatstroke (duration, 63-185 min). As compared with normothermic controls, all vehicle-pretreated heatstroke animals displayed higher levels of creatinine, serum urea nitrogen, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, tumor necrosis factor alpha, prothrombin time, activated partial thromboplastin time and D-dimer in the plasma, cellular ischemia and injury markers in striatum, and intracranial pressure. In contrast, all vehicle pretreated heatstroke animals had lower levels of mean arterial pressure, cerebral perfusion pressure, cerebral blood flow, brain Po2, and platelet count and protein C in the plasma. Immediately after the start of heat exposure, the previous administration of platonin significantly improved survival time by reducing the systemic inflammation, hypercoagulable state, and tissue ischemia and damage during heatstroke. The results demonstrate that platonin is effective for attenuation of heatstroke reactions. PMID- 17117137 TI - Pifithrin-alpha attenuates p53-mediated apoptosis and improves cardiac function in response to myocardial ischemia/reperfusion in aged rats. AB - Ischemic cardiovascular disease is a common age-related disease. The p53 dependent cardiac myocyte apoptosis induced by myocardial ischemia/reperfusion (MI/R) is an important feature in the progression of ischemic heart disease. In the present studies, we hypothesized that inhibition of p53-dependent myocyte apoptosis may improve cardiac dysfunction in aged rats after MI/R. A dose (2.2 mg/kg, i.p.) of pifithrin-alpha (PFT), a p53 inhibitor, or saline was administered to 20-month-old male F344 rats, which were subjected to 30 min of myocardial ischemia by ligating the left main coronary artery, followed by release of the ligature and 4 h of reperfusion. Results of our experiments indicate that MI/R induced a significant decrease in cardiac output index (CI) and mean arterial blood pressure (MABP). Administration of PFT to aged rats 40 min before ischemia significantly improved CI and MABP during 3 to 4 h of reperfusion. The improvement of cardiac function was associated with a marked reduction in DNA fragmentation in the area at risk of the heart when compared with aged MI/R rats pretreated with saline. Interestingly, treatment with PFT 10 min after ischemia or 10 min after reperfusion had a similar protective effect on CI and MABP, but this effect did not reach statistical significance when compared with aged MI/R rats pretreated with saline. Treatment with PFT, however, did not influence plasma creatine kinase activity and the number of circulating leukocytes and infiltrated leukocytes in the area at risk of the heart. Moreover, results of Western blot show that pretreatment with PFT significantly attenuated the ratio of Bax to Bcl-2 in the area-at-risk tissue of the heart compared with that of rats pretreated with saline. Our results suggest that pretreatment with PFT significantly improved cardiac function. The mechanism of protective effect of PFT may involve the inhibition of p53 transcriptional function, thereby attenuating the p53/Bax-mediated myocyte apoptosis during the reperfusion period. PMID- 17117138 TI - Effects of thoracic epidural anesthesia on hemodynamics and global oxygen transport in ovine endotoxemia. AB - Besides providing effective analgesia, thoracic epidural anesthesia (TEA) has been shown to decrease perioperative morbidity and mortality. Because of its vasodilatory properties in association with the sympathetic blockade, however, TEA may potentially aggravate cardiovascular dysfunctions resulting from sepsis and systemic inflammatory response syndrome. The objective of the present study was to assess the effects of TEA on hemodynamics, global oxygen transport, and renal function in ovine endotoxemia. After a baseline measurement in healthy sheep (n = 18), Salmonella typhosa endotoxin was centrally infused at incremental doses to induce and maintain a hypotensive-hypodynamic circulation using an established protocol. The animals were then randomly assigned to one of two groups. In the treatment group, continuous TEA was initiated with 0.1 mL.kg of 0.125% bupivacaine at the onset of endotoxemia and maintained with 0.1 mL.kg.h. In the control group, the same amount of isotonic sodium chloride solution was injected through the epidural catheter. In the animals surviving the entire experiment (n = 7 per group), cardiac index and mean arterial pressure decreased in a dose-dependent manner during endotoxin infusion. In the TEA group, neither systemic hemodynamics nor global oxygen transport were impaired beyond the changes caused by endotoxemia itself. Urinary output was increased in the TEA group as compared with the control group (P < 0.05). In this model of endotoxic shock, TEA improved renal perfusion without affecting cardiopulmonary hemodynamics and global oxygen transport. PMID- 17117139 TI - Intraperitoneal administration of hyperbarically oxygenated perfluorochemical enhances preservation of intestinal mucosa against ischemia/reperfusion injury. AB - Perfluorochemicals (PFCs) have a high solubility for oxygen. We have previously demonstrated the effect of peritoneal lavage with oxygenated PFC (O2-PFC) on ameliorating ischemia/reperfusion (I/R)-induced intestinal ischemic damage in an animal model. In this study, we applied hyperbarically O2-PFC (HBO-PFC) to investigate whether a larger amount of oxygen carried by PFC could enhance the protective effect of O2-PFC during intestinal malperfusion. Rats were subjected to ischemia by clamping the superior mesenteric artery (SMA) for 90 min. The SMA was then declamped. Rats were divided into four groups. In group A, only anesthesia and abdominal incision were performed. In group B, SMA was clamped without O2-PFC. In group C, during the SMA clamp, 1 atm O2-PFC was injected into the abdominal cavity. In group D, 5 atm O2-PFC (HBO-PFC) was prepared using a custom-made hyperbaric oxygen tank and administered to the abdominal cavity during the SMA clamp. Ileal tissue adenosine triphosphate (ATP) levels 90 min after SMA declamping were determined using luciferase assay. To assess intestinal mucosal barrier function at 90 min after release of the SMA clip, everted gut sacs were prepared to measure the mucosal-to-serosal passage of fluorescein conjugated dextran (FD4, molecular weight = 4 kDa). Thirty minutes after i.p. administration, partial pressure of oxygen in HBO-PFC remained around 1000 mmHg, whereas partial pressure of oxygen in 1 atm O2-PFC decreased to around 400 mmHg. The intestinal tissue ATP was significantly preserved in group D. Moreover, the mucosal hyperpermeability of the gut sac after I/R was significantly ameliorated in group D. Hyperbarically oxygenated perfluorochemical might supply a larger amount of oxygen to ischemic tissue during SMA clamp, which protected the small intestine from I/R injury, possibly caused by the maintenance of tissue ATP levels during ischemia. PMID- 17117140 TI - Sesame oil attenuates acute iron-induced lipid peroxidation-associated hepatic damage in mice. AB - Acute iron intoxication from the accidental ingestion of iron-containing preparations is one important cause of death in children. The aim of this study was to investigate the protective effect of sesame oil on acute iron-induced lipid peroxidation (LPO) and hepatic injury in mice. Acute iron intoxication was induced by giving ferric nitrilotriacetate to mice. Hepatic function was assessed using blood biochemistry. Free radicals were determined using a high-performance chemiluminescence analyzer. Ferric nitrilotriacetate increased serum ferrous (Fe) and LPO levels, and induced acute hepatic injury. Sesame oil (a) dose-dependently decreased acute iron-induced LPO and hepatic injury, (b) reduced acute iron associated hydroxyl radical and superoxide anion generation, and (c) inhibited the activity of xanthine oxidase in acute iron intoxication. Thus, sesame oil might ameliorate LPO and acute hepatic injury by inhibiting xanthine oxidase initiated superoxide anion generation, thereby reducing hydroxyl radical production, at least partially, in acutely iron-intoxicated mice. PMID- 17117141 TI - Cytokine-triggered decreases in levels of phosphorylated eukaryotic initiation factor 4G in skeletal muscle during sepsis. AB - Chronic septic abscess formation causes an inhibition of protein synthesis in gastrocnemius that is not observed in rats with a sterile abscess. The inhibition is associated with an impaired translation initiation. The present study was designed to investigate the effects of sepsis on the level of phosphorylated eukaryotic initiation factor (eIF) 4G in gastrocnemius after induction of a chronic intra-abdominal sterile or septic abscess as a possible mechanism to account for the impairment of translation initiation during sepsis. The extent of phosphorylated eIF4G was reduced by more than 50% (P< 0.05) and 68% (P < 0.01) in gastrocnemius after 3 and 5 days, respectively, and returned to control values after 14 days of abscess formation in septic rats compared with sterile inflammatory animals. To examine the mediators of the septic process contributing to the decreased levels of phosphorylated eIF4G, the cytokine response to sepsis was pharmacologically modulated. First, treatment of septic rats with tumor necrosis factor (TNF) binding protein or interleukin (IL) 1 receptor antagonist increased the level of phosphorylated eIF4G. Second, infusion of TNF-alpha for 24 h in control rats resulted in a 70% decrease in phosphorylated eIF4G. Third, infusion of IL-1ra led to an increase in the level of phosphorylation of eIF4G in rats infused with TNF-alpha. Taken together, the data indicate that a cytokine dependent decrease in the steady state phosphorylation of eIF4G is a possible mechanism accounting for the inhibition of skeletal muscle protein synthesis during sepsis. Furthermore, the findings support a role of IL-1 as the proinflammatory mediator responsible for the reduced level of phosphorylated eIF4G. PMID- 17117144 TI - Prognosis of AIDS-related systemic non-Hodgkin lymphoma treated with chemotherapy and highly active antiretroviral therapy depends exclusively on tumor-related factors. AB - OBJECTIVES: To assess complete remission (CR) and survival in patients with systemic AIDS-related non-Hodgkin lymphoma (ARL) receiving highly active antiretroviral therapy (HAART). METHODS: We analyzed the Grupo de Estudio del SIDA register of systemic ARL, which started in Jan 1994, to collect cases diagnosed at 15 institutions prospectively and with active follow-up every 6 months. The date of censorship for this study was March 2005. RESULTS: During the study period, 210 consecutive patients were diagnosed with ARL, with a median age 39 of years, 75.7% of whom were male, and with a median baseline CD4 count of 160 cells/microL. Histologic subtypes were diffuse large B-cell lymphoma (DLCL; n = 153 [72.9%]), Burkitt and atypical Burkitt/Burkitt-like lymphoma (BL; n = 40 [19.0%]), T-cell lymphoma (TC; n = 8 [3.8%]), and miscellaneous (n = 9 [4.3%]). Chemotherapy with or without other modalities was administered to 186 (88.6%) patients. In an intent-to-treat analysis of 184 patients who received at least 1 chemotherapy course with adequate follow-up to assess their response, 119 (64.7%) achieved CR, and the median length of survival (Kaplan-Meier analysis) was 52 months (95% confidence interval [CI]: 23 to 82 months). Factors independently associated with CR were histologic subtype and International Prognostic Index (IPI) score. Factors independently associated with improved overall length of survival (OS) were CR, low IPI score, and histologic subtype. The single factor independently associated with disease-free survival was Ann Arbor stage. CONCLUSIONS: In patients with ARL treated with HAART, CR was associated exclusively with tumor-related factors. The CR rate was poorer in patients with BL and TC subtypes and was inversely correlated with IPI score. OS was independently associated with CR, IPI score, and the histologic subtype. PMID- 17117145 TI - Selection and persistence of viral resistance in HIV-infected children after exposure to single-dose nevirapine. AB - BACKGROUND: Single-dose nevirapine (sd-NVP) is the mainstay of prevention of mother-to-child transmission programs in developing countries. Exposure to sd-NVP selects for resistance mutations, however. We longitudinally assessed these mutations in HIV-1-infected infants from Soweto and Durban, South Africa. METHODS: We prospectively followed 465 infants who received sd-NVP after enrolling their mothers when pregnant. If HIV infected, their virus was genotyped, using the ViroSeq HIV-1 Genotyping System, to detect resistant mutations. Those with resistance were genotyped at 6 months and then every 6 months out to 18 months if resistance was detected at the previous visit. RESULTS: Of 53 HIV-infected infants, 24 (45.3%) had detectable resistance at their first visit, when the most frequent mutations were Y181C (75%), K103N (25%), and Y188C (12%). Of those whose visit was before 12 weeks of age, 2 of 42 infants shared identical resistance mutations with their mothers. By 18 months of age, 11 of 24 infants with resistance had died and 1 still had the Y181C mutation. CONCLUSIONS: Resistant mutations were selected in half of the infants exposed to sd-NVP, but fewer were detected over time and, unlike the case in their mothers, Y181C dominated initially and persists. Transient resistance mutations may have a negative impact on highly active antiretroviral therapy in infants and children. PMID- 17117147 TI - Frailty in the elderly: the physical dimension. AB - Frailty may be considered as a vulnerable status, which can precede the onset of overt disability. Operational definitions of frailty vary widely according to the conceptual framework: some authors consider frailty in a broad sense, which encompasses the physical, social, cognitive, psychological dimensions and comorbidity, whereas others define the syndrome more restrictively, mainly on the basis of performance parameters, such as gait speed, grip strength and physical activity. All these definitions are provided of a high predictive value for adverse outcomes, such as disability, hospitalization and mortality. Sarcopenia (i.e. the reduction of muscular mass and function) plays a predominant role in the pathogenesis of frailty. Among the factors responsible for sarcopenia (such as proinflammatory cytokines, low growth hormone and testosterone levels, increased production of oxygen free radicals, malnutrition and reduced neurological drive), immobility and lack of exercise have a preponderant role. Therefore, the diagnosis of frailty is mandatory for the early identification of a subset of elderly subjects at high risk, which can receive benefit from rehabilitation. A self-report and objective evaluation of physical performance are the best indicators of frailty in elderly subjects, a poor performance suggesting the need of an early and proper intervention. Structured exercise programs are effective in contrasting the progression of frailty, but an healthy and active lifestyle may be sufficient for delaying the onset of disability. In conclusion, there is clear evidence for prescription of exercise within the mainstream of the medical practice, rather than as an optional adjunct to standard care of the oldest old, given the public health implication of frailty, whose prevalence is going to increase in western populations. PMID- 17117149 TI - Infrared-LAMP: two-photon uncaging and imaging of gap junctional communication in three dimensions. PMID- 17117150 TI - Complementary methods for studies of protein interactions in living cells. PMID- 17117151 TI - Single protein complex visualization: seeing is believing. PMID- 17117152 TI - G protein-coupled receptors: too many dimers? PMID- 17117153 TI - Atomic force microscopy of membrane proteins separating two aqueous compartments. PMID- 17117154 TI - QUICKstep and GS-TAP: new moves for protein-interaction analysis. PMID- 17117155 TI - Protein complex expression by using multigene baculoviral vectors. AB - Elucidation of the molecular basis of protein-interaction networks, in particular in higher eukaryotes, is hampered by insufficient quantities of endogenous multiprotein complexes. Present recombinant expression methods often require considerable investment in both labor and materials before multiprotein expression, and after expression and biochemical analysis these methods do not provide flexibility for expressing an altered multiprotein complex. To meet these demands, we have recently introduced MultiBac, a modular baculovirus-based system specifically designed for eukaryotic multiprotein expression. Here we describe new transfer vectors and a combination of DNA recombination-based methods, which further facilitate the generation of multigene cassettes for protein coexpression (Fig. 1), thus providing a flexible platform for generation of protein expression vectors and their rapid regeneration for revised expression studies. Genes encoding components of a multiprotein complex are inserted into a suite of compatible transfer vectors by homologous recombination. These progenitor constructs are then rapidly joined in the desired combination by Cre-loxP mediated in vitro plasmid fusion. Protocols for integration of the resulting multigene expression cassettes into the MultiBac baculoviral genome are provided that rely on Tn7 transposition and/or Cre-loxP reaction carried out in vivo in Escherichia coli cells tailored for this purpose. Detailed guidelines for multigene virus generation and amplification, cell culture maintenance and protein production are provided, together with data illustrating the simplicity and remarkable robustness of the present method for multiprotein expression using a composite MultiBac baculoviral vector. PMID- 17117158 TI - The oncofetal protein glypican-3 is a novel marker of hepatic progenitor/oval cells. AB - Glypican-3 (Gpc3), a cell surface-linked heparan sulfate proteoglycan is highly expressed during embryogenesis and is involved in organogenesis. Its exact biological function remains unknown. We have studied the expression of Gpc3 in fetal and adult liver, in liver injury models of activation of liver progenitor cells: D-galactosamine and 2-acetylaminofluorene (2-AAF) administration followed by partial hepatectomy (PH) (2-AAF/PH); and in the Solt-Farber carcinogenic model: by initiation with a single dose of diethylnitrosamine and promotion with 2-AAF followed by PH treatment. Gpc3 expression was studied using complementary DNA microarrays, reverse transcriptase-polymerase chain reaction, in situ hybridization (ISH); ISH combined with immunohistochemistry (IHC) and immunofluorescent microscopy. We found that Gpc3 is highly expressed in fetal hepatoblasts from embryonic days 13 through 16 and its expression gradually decreases towards birth. Dual ISH with Gpc3 and alpha-fetoprotein (AFP) probes confirmed that only hepatoblasts and no other fetal liver cells express Gpc3. At 3 weeks after birth the expression of Gpc3 mRNA and protein was hardly detected in the liver. Gpc3 expression was highly induced in oval cell of D-gal and 2 AAF/PH treated animals. Dual ISH/IHC with Gpc3 riboprobe and cytokeratin-19 (CK 19) antibody revealed that Gpc3 is expressed in activated liver progenitor cells. ISH for Gpc3 and AFP performed on serial liver sections also showed coexpression of the two-oncofetal proteins. FACS isolated oval cells with anti-rat Thy1 revealed expression of Gpc3. Gpc3 expression persists in atypical duct-like structures and liver lesions of animals subjected to the Solt-Farber model of initiation and promotion of liver cancer expressing CK-19. In this work we report for the first time that the oncofetal protein Gpc3 is a marker of hepatic progenitor cells and of early liver lesions. Our findings show further that hepatic progenitor/oval cells are the target for malignant transformation in the Solt-Farber model of hepatic carcinogenesis. PMID- 17117159 TI - Commonly used caspase inhibitors designed based on substrate specificity profiles lack selectivity. PMID- 17117160 TI - Heterotrimeric G protein alpha subunit is involved in rice brassinosteroid response. AB - Heterotrimeric G proteins are known to function as messengers in numerous signal transduction pathways. The null mutation of RGA (rice heterotrimeric G protein alpha subunit), which encodes the alpha subunit of heterotrimeric G protein in rice, causes severe dwarfism and reduced responsiveness to gibberellic acid in rice. However, less is known about heterotrimeric G protein in brassinosteroid (BR) signaling, one of the well-understood phytohormone pathways. In the present study, we used root elongation inhibition assay, lamina inclination assay and coleoptile elongation analysis to demonstrated reduced sensitivity of d1 mutant plants (caused by the null mutation of RGA) to 24-epibrassinolide (24-epiBL), which belongs to brassinosteroids and plays a wide variety of roles in plant growth and development. Moreover, RGA transcript level was decreased in 24-epiBL treated seedlings in a dose-dependent manner. Our results show that RGA is involved in rice brassinosteroid response, which may be beneficial to elucidate the molecular mechanisms of G protein signaling and provide a novel perspective to understand BR signaling in higher plants. PMID- 17117161 TI - Training for tomorrow's brain sciences. PMID- 17117162 TI - New practice parameters in Parkinson's disease. PMID- 17117163 TI - Secondary prevention measures after a stroke--should they target stroke or heart disease? PMID- 17117164 TI - Can amantadine therapy delay the onset of dementia in Parkinson's disease? PMID- 17117165 TI - Skin biopsy findings predict development of symptomatic neuropathy in patients with HIV. PMID- 17117166 TI - Management of bladder dysfunction in patients with multiple-system atrophy. PMID- 17117167 TI - Can medication overuse headache be treated by abrupt withdrawal of the overused agent? PMID- 17117168 TI - New screening tool for identifying major depression in patients with epilepsy. PMID- 17117169 TI - Are frontotemporal lobar degeneration, progressive supranuclear palsy and corticobasal degeneration distinct diseases? AB - New findings relating to the clinical, genetic and molecular bases of neurodegenerative disorders have led to a shift away from traditional nomenclatures of clinical syndromes. Historically, frontotemporal lobar degeneration (FTLD), corticobasal degeneration (CBD) and progressive supranuclear palsy (PSP) were classified on the basis of distinct clinical and pathological features. In recent years, however, advances in molecular and genetic research have led clinicians to suggest that the similar etiologies of the three disorders warrant their amalgamation into a single disorder with three subtypes. In this Review, we consider the utility and validity of combining FTLD, CBD and PSP. The earliest reports of these disorders demonstrate their distinctiveness, whereas recent findings challenge traditional nomenclatures by showing etiological overlap. For example, tau inclusions have been confirmed in patients with CBD and those with PSP, and in some patients with FTLD, implying that all three disorders are 'tauopathies'. Furthermore, most patients with progressive nonfluent aphasia, a subtype of FTLD, show PSP or CBD post-mortem. Even tau-related cases of FTLD, CBD and PSP are distinguishable on the basis of other criteria, however, and many FTLD cases do not show tau pathology. We argue, therefore, that FTLD, CBD and PSP should be considered as pathologically similar but distinct syndromes. New research criteria for CBD and PSP should note that progressive nonfluent aphasia is often a precursor of these conditions. PMID- 17117170 TI - Essential tremor: emerging views of a common disorder. AB - Essential tremor (ET) is the most prevalent tremor disorder. ET has traditionally been viewed as a monosymptomatic disorder characterized by a kinetic arm tremor, but this definition is gradually being replaced. The clinical spectrum has come to include several motor features, including tremor and ataxia, and several non motor features, including possible cognitive impairment and personality disturbances. Postmortem studies are revealing several different patterns of pathology. The emerging view is that ET might be a family of diseases, unified by the presence of kinetic tremor, but further characterized by etiological, clinical and pathological heterogeneity. Effective pharmacological treatments for the disorder remain limited, although new insights into disease mechanisms might result in more-effective therapies. In addition, recent investigations of environmental toxicants that might be linked to ET open the way towards primary disease prevention through a reduction in exposure to these factors. PMID- 17117171 TI - Mechanisms of Disease: astrocytes in neurodegenerative disease. AB - The term neurodegenerative disease refers to the principal pathology associated with disorders such as amyotrophic lateral sclerosis, Alzheimer's disease, Huntington's disease and Parkinson's disease, and it is presumed that neurodegeneration results in the clinical findings seen in patients with these diseases. Decades of pathological and physiological studies have focused on neuronal abnormalities in these disorders, but it is becoming increasingly evident that astrocytes are also important players in these and other neurological disorders. Our understanding of the normative biology of astrocytes has been aided by the development of animal models in which astrocyte-specific proteins and pathways have been manipulated, and mouse models of neurodegenerative diseases have also revealed astrocyte-specific pathologies that contribute to neurodegeneration. These models have led to the development of targeted therapies for pathways in which astrocytes participate, and this research should ultimately influence the clinical treatment of neurodegenerative disorders. PMID- 17117172 TI - How to bring neurology and psychiatry closer? Trainees, trainers and teachers support shared learning. PMID- 17117173 TI - Activity performance and upper extremity function in cervical spinal cord injury patients according to the Klein-Bell ADL Scale. AB - STUDY DESIGN: Cross-sectional study. OBJECTIVES: (1) To examine whether the Klein Bell ADL Scale (K-B Scale) discriminates cervical spinal cord injury (SCI) patients in daily activities and to explore its applicability in this group of patients. (2) To examine the association between basic ADL and upper extremity function. (3) To investigate if grip ability can be discerned in the scale. SETTING: Spinal Cord Injury Unit, Sahlgrenska University Hospital, Goteborg, Sweden. METHODS: Fifty-five patients with cervical SCI with no prior reconstructive hand surgery were included in the study. Analyses of the patient's independence were made according to the K-B Scale. Three additional analyses were carried out, the first examined whether the use of assistive devices and house and car adaptations influenced independence. The last two used different approaches to investigate whether arm and grip function could be detected in the K-B scale. RESULTS: Raw score in the K-B Scale can discriminate for independence in daily activities but the scale's weight scheme does not function for cervical SCI patients. Assistive devices and car and house adaptations can compensate for dependence in daily activities. Lack of grip function decreases the patient's ability to become independent. Diagnosis-related activities cannot be assessed in all items. CONCLUSION: The K-B Scale's raw score was useful assessing daily activities in cervical SCI patients. Its reliability in conjunction with arm and grip function in patients with cervical SCI has yet to be proven. PMID- 17117174 TI - A cadaveric histological investigation of the prostate with three-dimensional reconstruction for better results in continence and erectile function after radical prostatectomy. AB - In this study our aim is to increase the understanding of the prostate and related organs anatomy for better continence and erectile function results after urological surgery. Prostate and related organs were dissected from seven cadavers. After dissection, 165 serial sections with 300 microm thickness were derived at a 100 microm interval. The histological images were examined and imported to the computer. Three-dimensional (3D) remodeling had been performed. The findings were evaluated into three categories: macroscopic, microscopic and 3D reconstruction. Striated muscle fibers had been detected at the anterior fibromuscular stroma in histological sections. In 3D remodeling, urethra seemed to be a complete functional unit, beginning from the trigone up to the membranous urethra. The neurovascular bundles run under the pelvic fascia on both sides and go through to the bladder neck at 5 and 7 o'clock. Computer remodeling demonstrated that neurovascular structures had a close association with the bladder neck and the seminal vesicle. Computer program made it possible to rotate all 3D-reconstructed figures by 360 degrees and examine them from all possible angles. All reconstructed structures can be examined together at the same time or one by one. Surgeons must pay special attention to the continence area described as a single unit, beginning from trigone to the membranous urethra, during the surgery. Meticulous dissection of the neurovascular bundles, especially close to the seminal vesicles and bladder neck, during the radical prostatectomy is necessary. These reconstructions can be used for the educational purpose of medical students as well as the urology surgeons. PMID- 17117175 TI - Effect of treatment with epoetin beta on short-term tumour progression and survival in anaemic patients with cancer: A meta-analysis. AB - To assess the early effect of epoetin beta on survival and tumour progression in anaemic patients with cancer, data were pooled from nine randomised clinical trials comparing epoetin beta with placebo or standard care. Studies were not primarily designed to assess these end points. Follow-up was for treatment duration plus 4 weeks following therapy completion. All adverse events (AEs) were retrospectively reviewed blinded, for progression. Thromboembolic events were also assessed. Data analysis involved standard statistical tests. Overall, 1413 patients were included (epoetin beta, n = 800; control, n = 613; 56% haematological, and 44% solid). Median initial epoetin beta dose was 30,000 IU/week. Overall survival during months 0-6 was similar with epoetin beta and control (0.31 vs 0.32 deaths/patient-year). No increased mortality risk was seen with epoetin beta (relative risk (RR) 0.97, 95% CI: 0.69, 1.36; P = 0.87). There was a significantly reduced risk of rapidly progressive disease for epoetin beta (RR 0.78, 95% CI: 0.62, 0.99; P = 0.042). Epoetin beta was associated with a slightly higher frequency of thromboembolic events vs control (5.9% vs 4.2% of patients) but thromboembolic-related mortality was identical in both groups (1.1%). Epoetin beta provided a slight beneficial effect on tumour progression and did not impact on early survival or thromboembolic-related mortality. PMID- 17117176 TI - No link between viral findings in the prostate and subsequent cancer development. AB - In an investigation of 201 prostate tissue samples from patients with benign prostate hyperplasia that later progressed to prostate cancer and 201 matched controls that did not, there were no differences in the prevalence of adenovirus, herpesvirus, papilloma virus, polyoma virus and Candida albicans DNA. PMID- 17117177 TI - The status of CDKN2A alpha (p16INK4A) and beta (p14ARF) transcripts in thyroid tumour progression. AB - CDKN2A locus on chromosome 9p21 encodes two tumour suppressor proteins pl6INK4A, which is a regulator of the retinoblastoma (RB) protein, and p14ARF, which is involved in the ARF-Mdm2-p53 pathway. The aim of this study was to determine if CDKN2A gene products are implicated in differentiated thyroid carcinogenesis and progression. We used real-time quantitative RT-PCR and immunohistochemistry to assess both transcripts and proteins levels in 60 tumours specimens. Overexpression of p14ARF and pl6INK4A was observed in follicular adenomas, follicular carcinomas and papillary carcinomas, while downregulation was found in oncocytic adenomas compared to nontumoral paired thyroid tissues. These deregulations were statistically significant for pl6INK4a (P=0.006) in follicular adenomas and close to statistical significance for p14ARF in follicular adenomas (P=0.06) and in papillary carcinomas (P=0.05). In all histological types, except papillary carcinomas, we observed a statistically significant relationship between p14ARF and E2F1 (r=0.64 to 1, P<0.05). Our data are consistent with involvement of CDKN2A transcript upregulation in thyroid follicular tumorigenesis as an early event. However, these deregulations do not appear to be correlated to the clinical outcome and they could not be used as potential prognostic markers. PMID- 17117178 TI - Very low prevalence of germline MSH6 mutations in hereditary non-polyposis colorectal cancer suspected patients with colorectal cancer without microsatellite instability. AB - Hereditary non-polyposis colorectal cancer (HNPCC) is caused by mutations in one of the mismatch repair genes MLH1, MSH2, MSH6, or PMS2 and results in high-level microsatellite instability (MSI-high) in tumours of HNPCC patients. The MSI test is considered reliable for indicating mutations in MLH1 and MSH2, but is questioned for MSH6. Germline mutation analysis was performed in 19 patients with an MSI-high tumour and absence of MSH2 and/or MSH6 protein as determined by immunohistochemistry (IHC), without an MLH1 or MSH2 mutation, and in 76 out of 295 patients suspected of HNPCC, with a non-MSI-high colorectal cancer (CRC). All 295 non-MSI-high CRCs were analysed for presence of MSH6 protein by IHC. In 10 patients with an MSI-high tumour without MSH2 and/or MSH6 expression, a pathogenic MSH6 mutation was detected, whereas no pathogenic MSH6 mutation was detected in 76 patients with a non-MSI-high CRC and normal MSH6 protein expression. In none of the 295 CRCs loss of MSH6 protein expression was detected. The prevalence of a germline MSH6 mutation is very low in HNPCC suspected patients with non-MSI-high CRC. Microsatellite instability analysis in CRCs is highly sensitive to select patients for MSH6 germline mutation analysis. PMID- 17117179 TI - Indoleamine 2,3-dioxygenase is a novel prognostic indicator for endometrial cancer. AB - Indoleamine 2,3-dioxygenase (IDO) is a tryptophan-catabolising enzyme inducing immune tolerance. The present study aimed to investigate IDO expression and its prognostic significance in endometrial cancer. Indoleamine 2,3-dioxygenase expression in endometrial cancer tissues (n = 80) was immunohistochemically scored as four groups (IDO-, 1+, 2+, and 3+). The high IDO expression (IDO2+ or 3+) in tumour cells was found in 37 (46.3%) of the 80 cases, and was positively correlated with surgical stage, myometrial invasion, lymph-vascular space involvement, and lymph node metastasis, but not with the histological grade. Patients with high IDO expression had significantly impaired overall survival and progression-free survival (PFS) (P = 0.002 and P = 0.001, respectively) compared to patients with no or weak expression of IDO (IDO- or 1+). The 5-year PFS for IDO-/1+, 2+, and 3+ were 97.7, 72.9, and 36.4%, respectively. Even in patients with early-stage disease (International Federation of Gynecology and Obstetrics I/II, n = 64), the PFS for IDO2+/3+ was significantly poor (P = 0.001) compared to that for IDO-/1+. On multivariate analysis, IDO expression was an independent prognostic factor for PFS (P = 0.020). These results indicated that the high IDO expression was involved in the progression of endometrial cancer and correlated with the impaired clinical outcome, suggesting that IDO is a novel and reliable prognostic indicator for endometrial cancer. PMID- 17117180 TI - HapMap-based study of the 17q21 ERBB2 amplicon in susceptibility to breast cancer. AB - ERBB2 is frequently amplified in breast tumours as part of a wide region of amplification on chromosome 17q21. This amplicon contains many candidate genes for breast cancer susceptibility. We used a genetic association study design to determine if common genetic variation (frequency>or=5%) in a 400-kb region surrounding ERBB2 and containing the PPARBP, CRK7, NEUROD2, PPP1R1B, STARD3, TCAP, PNMT, CAB2, ERBB2, C17ORF37, GRB7 and ZNFN1A3 genes, was associated with breast cancer risk. Sixteen tagging single-nucleotide polymorphisms (tSNPs) selected within blocks of linkage disequilibrium from the HapMap database, one HapMap singleton SNP, and six additional SNPs randomly selected from dbSNP were genotyped using Taqman in a large study set of British women (2275 cases, 2280 controls). We observed no association between any of the genotypes or associated haplotypes and disease risk. In order to simulate unidentified SNPs, we performed the leave-one-out cross-validation procedure on the HapMap data; over 90% of the common genetic variation was well represented by tagging polymorphisms. We are therefore likely to have tagged any common variants present in our population. In summary, we found no association between common genetic variation in the 17q21 ERBB2 amplicon and breast cancer risk in British women. PMID- 17117181 TI - Evidence-based guidelines and decision support services: A discussion and evaluation in triple assessment of suspected breast cancer. AB - Widespread health service goals to improve consistency and safety in patient care have prompted considerable investment in the development of evidence-based clinical guidelines. Computerised decision support (CDS) systems have been proposed as a means to implement guidelines in practice. This paper discusses the general concept in oncology and presents an evaluation of a CDS system to support triple assessment (TA) in breast cancer care. Balanced-block crossover experiment and questionnaire study. One stop clinic for symptomatic breast patients. Twenty four practising breast clinicians from United Kingdom National Health Service hospitals. A web-based CDS system. Clinicians made significantly more deviations from guideline recommendations without decision support (60 out of 120 errors without CDS; 16 out of 120 errors with CDS, P < 0.001). Ignoring minor deviations, 16 potentially critical errors arose in the no-decision-support arm of the trial compared with just one (P = 0.001) when decision support was available. Opinions of participating clinicians towards the CDS tool became more positive after they had used it (P < 0.025). The use of decision support capabilities in TA may yield significant measurable benefits for quality and safety of patient care. This is an important option for improving compliance with evidence-based practice guidelines. PMID- 17117182 TI - High sustained efficacy of a prophylactic quadrivalent human papillomavirus types 6/11/16/18 L1 virus-like particle vaccine through 5 years of follow-up. AB - Human papillomavirus (HPV) causes cervical, vulvar, and vaginal cancers, precancerous dysplasia, and genital warts. We report data for the longest efficacy evaluation to date of a prophylactic HPV vaccine. In total, 552 women (16-23 years) were enrolled in a randomised, placebo-controlled study of a quadrivalent HPV 6/11/16/18 L1 virus-like-particle vaccine with vaccination at months 0, 2, and 6. At regular intervals through 3 years, subjects underwent gynaecologic examination, cervicovaginal sampling for HPV DNA, serum anti-HPV testing, and Pap testing, with follow-up biopsy as indicated. A subset of 241 subjects underwent two further years of follow-up. At 5 years post enrollment, the combined incidence of HPV 6/11/16/18-related persistent infection or disease was reduced in vaccine-recipients by 96% (two cases vaccine versus 46 placebo). There were no cases of HPV 6/11/16/18-related precancerous cervical dysplasia or genital warts in vaccine recipients, and six cases in placebo recipients (efficacy = 100%; 95% CI:12-100%). Through 5 years, vaccine-induced anti-HPV geometric mean titres remained at or above those following natural infection. In conclusion, a prophylactic quadrivalent HPV vaccine was effective through 5 years for prevention of persistent infection and disease caused by HPV 6/11/16/18. This duration supports vaccination of adolescents and young adults, which is expected to greatly reduce the burden of cervical and genital cancers, precancerous dysplasia, and genital warts. PMID- 17117183 TI - Authenticity and drug resistance in a panel of acute lymphoblastic leukaemia cell lines. AB - Cell lines are important models for drug resistance in acute lymphoblastic leukaemia (ALL), but are often criticised as being unrepresentative of primary disease. There are also doubts regarding the authenticity of many lines. We have characterised a panel of ALL cell lines for growth and drug resistance and compared data with that published for primary patient specimens. In contrast to the convention that cell lines are highly proliferative, those established in our laboratory grow at rates similar to estimates of leukaemic cells in vivo (doubling time 53-442 h). Authenticity was confirmed by genetic fingerprinting, which also demonstrated the potential stability of long-term cultures. In vitro glucocorticoid resistance correlated well with that measured ex vivo, but all lines were significantly more sensitive to vincristine than primary specimens. Sensitivity to methotrexate was inversely correlated to that of glucocorticoids and L-asparaginase, indicating possible reciprocity in resistance mechanisms. A cell line identified as highly methotrexate resistant (IC50 > 8000-fold higher than other lines) was derived from a patient receiving escalating doses of the drug, indicating in vivo selection of resistance as a cause of relapse. Many of these lines are suitable as models to study naturally occurring resistance phenotypes in paediatric ALL. PMID- 17117185 TI - Survival of death certificate initiated registrations: Selection bias, incomplete trace-back or higher mortality? AB - Cases first notified to a Registry and successfully followed back have an apparently worse prognosis than cases registered in life. A simple approach can be used to assess whether this is due to selection bias, incomplete follow-back or intrinsically higher mortality. For the colorectal, breast and stomach cancers studied and for comparable registries, the main explanations are likely to be selection bias and higher mortality. PMID- 17117184 TI - First international consensus on the methodology of lymphangiogenesis quantification in solid human tumours. AB - The lymphatic system is the primary pathway of metastasis for most human cancers. Recent research efforts in studying lymphangiogenesis have suggested the existence of a relationship between lymphatic vessel density and patient survival. However, current methodology of lymphangiogenesis quantification is still characterised by high intra- and interobserver variability. For the amount of lymphatic vessels in a tumour to be a clinically useful parameter, a reliable quantification technique needs to be developed. With this consensus report, we therefore would like to initiate discussion on the standardisation of the immunohistochemical method for lymphangiogenesis assessment. PMID- 17117186 TI - Impact of beta2-adrenoreceptor gene variants on cardiac cavity size and systolic function in idiopathic dilated cardiomyopathy. AB - In heart failure, the Arg16Gly and Gln27Glu polymorphisms of the beta2 adrenoreceptor (beta2-AR) gene are associated with exercise-capacity, clinical outcomes and response to beta-AR blocker therapy. Whether beta2-AR gene variants mediate these effects in-part through an impact on cardiac structural remodeling and pump function independent of the effects of beta-blockers is uncertain. We evaluated whether the Arg16Gly and Gln27Glu variants of the beta2-AR gene predict left ventricular ejection fraction (LVEF) and LV end diastolic diameter (LVEDD) in patients with idiopathic dilated cardiomyopathy (IDC) before and 6 months after receiving standard medical therapy other than beta-AR blockers. In all, 394 patients with IDC and 393 age and gender-matched controls were genotyped for the beta2-AR gene variants using restriction-fragment length polymorphism-based techniques. LVEF and dimensions were determined in 132 patients (of whom 71 were newly diagnosed) both at baseline and after 6 months. Genotype of neither variant was associated with the presence of IDC. Moreover, beta2-AR genotype did not determine LVEF or LV dimensions prior to initiating therapy. After 6 months of therapy, LVEF increased by 7.1+/-1.0 absolute units (P<0.0001) and LVEDD decreased by 0.27+/-0.06 cm (P<0.02). Adjusting for baseline values as well as gender, age, and type of angiotensin-converting enzyme inhibitor therapy received, genotype was associated with neither final LVEF and LVEDD, nor change in LVEF and LVEDD. In conclusion, these data suggest that in heart failure, the functional Arg16Gly and Gln27Glu variants of the beta2-AR gene have no independent effect on adverse structural remodeling and pump function. PMID- 17117187 TI - Differential association of adiponectin with cardiovascular risk markers in men and women? The KORA survey 2000. AB - BACKGROUND: In men, high adiponectin concentrations were related to a lower risk of myocardial infarction, whereas no association with cardiovascular events was found in women. OBJECTIVE: To investigate sex differences in the associations of adiponectin with cardiovascular risk factors. DESIGN: Cross-sectional population based KORA Survey 2000 in Southern Germany using the same study methods for cardiovascular risk factors as the former WHO MONICA project. PARTICIPANTS: A total of 697 men and 657 women, aged 55-74 years. Glucose tolerance status was assessed by oral glucose tolerance tests. RESULTS: Adiponectin (geometric mean, interquartile range; microg/ml) levels were significantly higher in women (11.1; 8.5-14.9) than in men (7.1; 5.2-9.6) (P<0.05). In univariate analyses, HDL cholesterol and age were significantly positively correlated with adiponectin in both sexes. Negative correlations were observed with BMI, waist circumference, fasting and postchallenge glucose, insulin, HOMA-IR, HbA1c, triglycerides, uric acid and CRP (P<0.01). In sex-specific multivariate regression, age and HDL cholesterol were independently positively, and fasting insulin and 2-h glucose were negatively related to adiponectin in both sexes. Uric acid was significantly inversely related to adiponectin in women only (sex interaction: P=0.02). Exploratory sex-specific factor analysis of adiponectin and the core components of the metabolic syndrome yielded four similar factors. Adiponectin loaded negatively on the 'lipids' factor in both sexes. CONCLUSION: The associations of adiponectin with cardiovascular risk factors showed a similar pattern in both sexes, except for uric acid. This small sex difference may not explain previous conflicting results on the association of adiponectin with cardiovascular events in men and women. PMID- 17117189 TI - [Suicidal behavior--what should be done?]. PMID- 17117188 TI - [Swindle and success]. PMID- 17117190 TI - [Skepticism against vaccination--paradox of preventive work]. PMID- 17117191 TI - [Incidence of breast cancer before and after implementation of mammography screening]. AB - BACKGROUND: A nationwide governmentally founded breast cancer screening program for women aged 50 - 69 years was gradually implemented in all Norwegian counties between November 1995 and February 2004 (the Norwegian Breast Cancer Screening Program, NBCSP). Such an implementation is expected to give an initial increased incidence followed by a decrease. We here present general and stage-specific incidences of breast cancer by age groups, in the first 9 counties that implemented the NBCSP. MATERIAL AND METHOD: The incidence of invasive breast cancer is shown for the counties Akershus, Hordaland, Oslo and Rogaland (AHOR), and Agder (east and west), Telemark, Troms and Finnmark (AgTmTF) for the period 1990-2003. AHOR started the NBCSP in 1995/1996, and AgTmTF started in 1999/2000. The incidence is based on recordings in the main database at the Cancer Registry, and is shown in rates per 100,000 women years. RESULTS: The incidence of breast cancer in women aged 50-69 years increased from 200 to 350 per 100,000 women years at start-up of the NBCSP and subsequently decreased to approximately 250. Before starting the program, the incidence was higher in women aged 70-74 years than in those aged 50 - 69. This changed in the subsequent period. During 1990 2003, the incidence of breast cancer was stable in women aged 20-49 and in those older than 74 years. At start-up of the NBCSP, a higher proportion of breast cancers in Stadium I were diagnosed as compared to the period before, especially among women aged 50-69 years. INTERPRETATION: Introduction of the NBCSP has changed the incidence of breast cancer in women aged 50-69 and in those aged 70 74 years. PMID- 17117192 TI - [Diffuse axonal injury in traumatic brain injury]. AB - BACKGROUND: Head trauma of varying severity may induce diffuse axonal injury. More attention is now given to this important type of injury, as examinations of head-injured patients with MRI have given us more knowledge. MATERIAL AND METHODS: We present a review of diffuse axonal injury with the main focus on clinical presentation and radiology, based on a Pubmed search and own experience. RESULTS AND INTERPRETATION: Axons seldom rupture at the moment of injury. It is more common that it takes hours or a few days until the axons are detached. Areas most commonly affected are white matter in the hemispheres, corpus callosum and the brain stem. Half of the patients with severe head injury have diffuse axonal injury, but this type of injury also occurs in patients with moderate and mild head injury. The clinical presentation and prognosis will therefore vary. Diffuse axonal injury can present with typical signs revealed by CT, but the CT scan may also be normal, especially when there is no bleeding. New MRI techniques are more sensitive and show that diffuse axonal injury occurs more often than previously assumed. MRI is therefore necessary to give the patients correct diagnoses and adequate rehabilitation and follow-up. PMID- 17117193 TI - [Suicidal behaviour in borderline personality disorder]. AB - Patients with borderline personality disorder have a substantially increased risk of self-destructive and suicidal behaviour, and are often seen as difficult to treat efficiently. A strong over-sensitivity to negative happenings in life, affective instability and impulsiveness are important risk factors for repetitive suicidal crises. Clinicians need to assess the suicide risk for individuals in general and more situation-specific risks. If there is a high risk, short term hospitalisation should be considered. Most patients will need long-term specialised treatment rather than a series of shorter treatments that might well worsen the prognosis. Several treatment models are currently available; among them are Dialectical Behaviour Therapy and Mentalisation-Based Therapy. The paper discusses how these treatment models address suicidal behaviour in patients with borderline personality disorder and the results that have been achieved in studies of treatment effectiveness. PMID- 17117194 TI - [Health-related use of the Internet in the Norwegian population]. AB - BACKGROUND: The use of the Internet for health purposes increases in the Norwegian population, more in some demographic groups than in others. In this questionnaire-based study, we explore the use of the Internet for such purposes. MATERIAL AND METHOD: 1007 Norwegians aged 15 years and older were interviewed by telephone in October 2005. RESULTS: 58% of the respondents in 2005 had used the Internet for health purposes, compared to 31% in 2001. Having visited the GP last year, being female, being young, living in a urban area, and having a white collar occupation were positively related to the use of the Internet for health purposes. 37% of the respondents considered the Internet to be an important or very important source of health information. 72% considered face-to-face communication with health care personnel to be important or very important. Nearly a quarter of the users (23%) reported that they had felt reassured by health information found on the net, whereas 10 % experienced increased anxiety from the same type of information. CONCLUSIONS: Norwegians' use of the Internet for health purposes continues to grow, but doctors and other health care personnel remain the most important sources of health information in the Norwegian population. PMID- 17117195 TI - [Are z-hypnotics better and safer sleeping pills than benzodiazepines?]. AB - Benzodiazepines and the benzodiazepine-like z-hypnotics (zopiclone, zolpidem, and zaleplon) have the same mode of action and many of the same effects. The use of z hypnotics has had a steady and large increase since their introduction in Norway, and sales data suggest extensive use among the elderly. The relatively short half lives of these drugs may cause less hangover effects, but z-hypnotics are hardly more effective or safer than benzodiazepines. The two classes of drugs should be prescribed with similar caution. PMID- 17117196 TI - [DNA--the future in vaccine technology?]. AB - DNA vaccines represent a new and promising technology that uses DNA to encode the antigen(s) of interest, instead of inoculating with attenuated or inactivated microbes or isolated antigens. Antigen is produced within the transfected cells minaicking a real life viral infection. This vaccine modality has been shown to elicit strong cellular immune responses and is promising for treating diseases where traditional vaccine approaches have failed. In spite of promising results in small animal models, DNA vaccines have so far proven less potent in human clinical trials. In this review we provide a general overview on the mechanisms of action for DNA vaccines, discuss potential benefits of traditional vaccine approaches and review current strategies for improving the immunogenicity of DNA vaccines to enable the successful transfer of the technology can be successfully transferred from mice to men. PMID- 17117197 TI - [Cancer vaccines]. AB - Immunotherapy is in principle attractive, as it exploits the immune system's own method of recognizing and destroying tumours. Tumour cells express a number of mutated or over-expressed antigens, which make it possible for the immune system to distinguish between tumour cells and normal cells. Cancer vaccines use tumour associated antigens to stimulate the patient's immune cells. In the clinical setting, individual differences between cancer patients is of vital importance, as the majority of tumour antigens may be unique to each individual. Genetic differences in patients' immune systems also suggest the need for individualised vaccines. This article gives a short introduction to the basic principles of tumour immunology and development of cancer vaccines. Clinical trials with emphasis on those with peptide and dendritic cell vaccines are presented, and the future development of cancer vaccines is discussed. A number of trials have shown tumour-specific immune responses and demonstrated that side-effects are generally not a problem. Limited documentation is available for the clinical effects. It is important to establish why some immune responses appear to give tumour regression while others are without clinical importance. PMID- 17117198 TI - [Vaccines against substance abuse]. PMID- 17117199 TI - [Psychological treatment of burnout]. PMID- 17117200 TI - [Pillory after scientific fraud--will it help?]. PMID- 17117201 TI - [Generic preparations--undocumented effects?]. PMID- 17117203 TI - [Dosaging of proton pumps inhibitors]. PMID- 17117204 TI - [Muscular tension in legs--a neglected problem]. PMID- 17117205 TI - [Cholera and vaccine]. PMID- 17117209 TI - [Wedding and anthrax]. PMID- 17117211 TI - In vitro co-stimulation of anti-tumor activity by soluble B7 molecules. AB - In order to investigate the anti-tumor activity of a soluble B7-1/immunoglobulin G fusion protein and explore an effective method to eliminate immune escape of tumor cells, a recombinant vector encoding this fusion protein was constructed and constitutively expressed in Chinese hamster ovary cells. After purification with protein G affinity chromatography, the soluble fusion protein was tested for bioactivity. Results showed that the fusion protein could significantly increase the density of B7-1 molecules on WEHI-3 cells, a mouse leukemia cell line. Through allogeneic mixed lymphocyte tumor cultures, it was demonstrated that, with the presence of the first signal, it could also significantly enhance T cell activation and killing activity against WEHI-3 cells and interleukin-2 secretion by activated mouse T lymphocytes. The conclusion can be drawn that the soluble B7 IgG fusion protein has a potent capacity to generate or enhance anti-tumor immune response in vitro, and its clinical value deserves further investigation. PMID- 17117212 TI - Sequence-specific Ni(II)-dependent peptide bond hydrolysis in a peptide containing threonine and histidine residues. AB - Previously we demonstrated that Ni(II) complexes of Ac-Thr-Glu-Ser-His-His-Lys NH2 hexapeptide, representing residues 120-125 of human histone H2A, and some of its analogs undergo E-S peptide bond hydrolysis. In this work we demonstrate a similar coordination and reactivity pattern in Ni(II) complexes of Ac-Thr-Glu-Thr His-His-Lys-NH2, its threonine analogue, studied using potentiometry, electronic absorption spectroscopy and HPLC. For the first time we present the detailed temperature and pH dependence of such Ni(II)-dependent hydrolysis reactions. The temperature dependence of the rate of hydrolysis yielded activation energy E(a) = 92.0 kJ mol(-1) and activation entropy DeltaS# = 208 J mol(-1) K(-1). The pH profile of the reaction rate coincided with the formation of the four-nitrogen square-planar Ni(II) complex of Ac-Thr-Glu-Thr-His-His-Lys-NH2. These results expand the range of protein sequences susceptible to Ni(II) dependent cleavage by those containing threonine residues and permit predictions of the course of this reaction at various temperatures and pH values. PMID- 17117213 TI - Ligand-centred reactivity in diiminepyridine complexes. AB - The diiminepyridine ligand, made famous by Brookhart and Gibson a decade ago through its use in the first iron-containing olefin polymerization catalyst, also displays an amazing variety of ligand-centred reactions, including electron transfer, alkylation, dimerization and deprotonation. The present Perspective summarizes these reactions and also speculates on the links between ligand- and metal-centred reactivity. PMID- 17117214 TI - Solution and solid state studies on the binding of isomeric carboranes C2B10H12 by p-Bu(t)-calix[5]arene. AB - p-Bu(t)-calix[5]arene forms crystalline inclusion complexes with o- and m carboranes in toluene or dichloromethane-hexane, but not with the p-isomer, the extended structures being based on 1 : 1 host-guest supermolecules, with the p Bu(t)-substituents creating a snug fit for o- and m-carborane; p-carborane forms a highly hexane soluble complex, induced by grinding, which crystallizes as fibres. Solution phase studies showed the presence of a 1 : 1 host-guest stoichiometry with all three isomeric carboranes as determined from Job plots. The association constants for the o- and m-carborane complexes are 6.4 +/- 0.3 M( 1) and 3.8 +/- 0.1 M(-1) respectively, whereas the p-isomer is only weakly associated. Competition experiments involving all three isomers show rapid exchange on the NMR time scale, and no selectivity in solution is evident. Selective association involving the o- and m-isomers in the solid state is therefore remarkable, and it is a manifestation of crystal packing forces which embodies the differences in dipole moments of the carboranes. PMID- 17117215 TI - Oxygen insertion in a carbon-phosphorus bond of the phenylethynyl-di-(tert-butyl) phosphine bridged dicobalt complex: exploring the nature of oxygen migration using DFT. AB - In the process of isolation under aerobic conditions phenylethynyl-di-(tert butyl)-phosphine bridged dicobalt complex [(micro PPh(2)CH(2)PPh(2))Co2(CO)4(micro,eta-PhC[triple bond]CP(t-Bu)2)] 4a underwent a partial oxidation. The identity of the oxidized product, [(micro PPh(2)CH(2)PPh(2))Co2(CO)4(micro,eta-PhC[triple bond]C-O-P([double bond]O)(t Bu)2)] 5, was established by spectroscopic means as well as the single-crystal X ray diffraction method. This is the first crystallographic evidence that unambiguously supports the formation of an organometallic version of a phosphinate ester. The mechanism for the formation of 5 from 4a was proposed, and its validity was examined by DFT means. For the purpose of comparison, a similar mechanism illustrating the transformation of PhC[triple bond]CP(t-Bu)2 1O into PhC[triple bond]C-O-P([double bond]O)(t-Bu)2 5O, the organic counterpart of 5, was examined by the same method. It was found that the metal fragment is indeed capable of assisting the oxidation process by lowering the activation energy, although the effect is small. The impact of the presence of an electron withdrawing substituent such as a fluorine atom in the alkynylphosphine was also investigated. Results demonstrated that the conversion of fluorine-substituted phosphines to the corresponding phosphinate esters can be achieved more readily. In addition, the energy barrier for the reaction of a phosphine with dioxygen yielding the phosphine oxide was calculated to be much lower than that on the way to the phosphinate ester. PMID- 17117216 TI - A molecular mechanics approach to mapping the conformational space of diaryl and triarylphosphines. AB - A molecular mechanics force field has been developed which accurately reproduces experimental solid state structures and conformer interconversion barriers for a series of sterically congested diaryl and triaryl phosphines and some of their chalcogenide and Cr(CO)5 derivatives. PMID- 17117217 TI - Metal-metal bonding in molecular actinide compounds: electronic structure of [M2X8](2-) (M = U, Np, Pu; X = Cl, Br, I) complexes and comparison with d-block analogues. AB - Density functional and multiconfigurational (ab initio) calculations have been performed on [M(2)X(8)](2-) (X = Cl, Br, I) complexes of 4d (Mo, Tc, Ru), 5d (W, Re, Os), and 5f (U, Np, Pu) metals in order to investigate general trends, similarities and differences in the electronic structure and metal-metal bonding between f-block and d-block elements. Multiple metal-metal bonds consisting of a combination of sigma and pi interactions have been found in all species investigated, with delta-like interactions also occurring in the complexes of Tc, Re, Np, Ru, Os, and Pu. The molecular orbital analysis indicates that these metal metal interactions possess predominantly d(z2) (sigma), d(xz) and d(yz) (pi), or d(xy) and d(x2-y2) (delta) character in the d-block species, and f(z3) (sigma), f(z2x) and f(z2y) (pi), or f(xyz) and f(z) (delta) character in the actinide systems. In the latter, all three (sigma, pi, delta) types of interaction exhibit bonding character, irrespective of whether the molecular symmetry is D(4h) or D(4d). By contrast, although the nature and properties of the sigma and pi bonds are largely similar for the D(4h) and D(4d) forms of the d-block complexes, the two most relevant metal-metal delta-like orbitals occur as a bonding and antibonding combination in D(4h) symmetry but as a nonbonding level in D(4d) symmetry. Multiconfigurational calculations have been performed on a subset of the actinide complexes, and show that a single electronic configuration plays a dominant role and corresponds to the lowest-energy configuration obtained using density functional theory. PMID- 17117218 TI - Group 4 catalysts for ethene polymerization containing tetradentate salicylaldiminato ligands. AB - The structural properties of three new classes of titanium and zirconium complex bearing tetradentate salicylaldiminato proligands are elucidated using X-ray diffraction and NMR spectroscopy. On activation with MAO co-catalyst, their behaviour in ethene polymerization depends strongly on the nature of the structure and the substitution pattern. One titanium complex based on a 2 aminobenzylamine (C3-chain) backbone has a trans arrangement of the co-ligand sites and, unsurprisingly, does not polymerize ethene. The 1,8-diaminonaphthalene (C3-chain) backbone gives a rather ring-strained cis complex, but was also unproductive. A range of cis complexes of zirconium with the 2,2'-diaminobibenzyl (C6-chain) backbone give low to moderate productivities of multimodal poly(ethene), while in contrast the structurally analogous titanium compounds provide highly active, single site catalysts. Thermal degradation of these catalysts is slowed significantly by a substitution pattern on the phenolate unit which sterically protects the imine donor unit; a phenomenon which has been previously observed in much lower activity catalysts based on 2,2' diaminobiphenyl (C4-chain) but which does not improve the stability of the very highly active unbridged systems. PMID- 17117219 TI - The role of halogenated carborane monoanions in olefin hydrogenation catalysed by cationic iridium phosphine complexes. AB - Iridium hydridophosphine complexes of general formula [Ir(PR3)2H2(anion)](PR3= PPh3, PMe2Ph; anion =[1-closo-CB(11)H(6)Cl(6)]-, [1-closo-CB(11)H(6)I(6)]-, [BAr(F)4]-) have been prepared by hydrogenation of cyclooctadiene precursor complexes. Solid-state structures of selected examples of these complexes reveal intimate contacts between the carborane anion and cation, with the anion binding through two lower-hemisphere halogen ligands. In CD2Cl2 solution the very weakly coordinating anions [1-closo-CB(11)H(6)Cl(6)]- and [BAr(F)4]- are suggested to favour the formation of solvent complexes such as [Ir(PR3)2H2(solvent)n][anion], while the [1-closo-CB(11)H(6)I(6)]- anion forms a tightly bound complex with the cationic iridium fragment. Calculated DeltaG values for anion reorganisation in d8-toluene reflect this difference in interaction between the anions and cation. With the bulky anion [1-closo-CB(11)Me(5)I(6)]- different complexes are formed: Ir(PPh3)H2(1-closo-HCB(11)Me(5)I(6)) and [(PPh3)3Ir(H2)H2][1-closo HCB(11)Me(5)I(6)] which have been characterised spectroscopically. Diffusion measurements in CD2Cl2 are also consistent with larger, solvent coordinated, complexes for the more weakly coordinating anions and a tighter interaction between anion and cation for [1-closo-CB(11)H(6)I(6)]-. All the complexes show some ion-paring in solution. Comparison with data previously reported for the [1 closo-CB(11)H(6)Br(6)]- anion shows that this anion--as expected--fits between [1 closo-CB(11)H(6)Cl(6)]- and [1-closo-CB(11)H(6)I(6)]- in terms of coordinating ability. Although not coordinating, the large [1-closo-CB(11)H(6)Cl(6)]- and [BAr(F)4)]- anions do provide some stabilisation towards the metal centre, as decomposition to the hydride bridged dimer [Ir2(PPh3)4H5]+ is retarded. This is in contrast to the [PF6]- salt where decomposition is immediate. As expected, complexes with the smaller phosphine PMe2Ph form tighter interactions with the carborane anions. These observations on the interaction between anion and cation in solution are reflected in benchmark hydrogenation studies that show a significant attenuation in rate of hydrogenation of cyclohexane on using the [1 closo-CB(11)H(6)I(6)]- anion or complexes with the PMe2Ph phosphine. We also comment on the reusability of the catalysts and their tolerance to water and oxygen impurities. Overall the catalyst with the [1-closo-CB(11)H(6)Br(6)]- anion shows the best combination of rate of hydrogenation, reusability and tolerance to impurities. PMID- 17117220 TI - Structure of sodium bis(N-methyl-iminodiacetato)iron(III): trans-meridional N coordination in the solid state and in solution. AB - The results of a detailed solid state and solution structural study of the Fe(III) bis-mida complex [Fe(III)(mida)(2)]- (mida = N-methyl-iminodiacetate) are reported. The structure of the sodium salt Na[Fe(mida)2][NaClO4]2.3H2O (1) was determined by single-crystal X-ray analysis. The complex anion in 1 contains a six-coordinate Fe(III) centre bound to two tridentate mida ligands arranged in the meridional configuration, and the mer Fe(III)N2O4 chromophore shows a high degree of distortion from regular octahedral symmetry. Raman- and UV/VIS/NIR spectroscopic measurements showed that no gross changes take place in the Fe(III) coordination sphere upon redissolution in water. Quantum chemical calculations of all three possible configurations of the [Fe(mida)2]- complex ion in the gas phase support the finding that the mer isomer is more stable than the u-fac (cis) and s-fac (trans) isomers. Redox potential measurements of the Fe(III/II)(mida) couple in dependence of pH led to the following values for the equilibrium contants: log beta(III)(101) = 11.98 +/- 0.05, log beta(III)(102) = 20.49 +/- 0.01, pK(III)(a1 OH) = 7.81; log beta(II)(101) = 6.17 +/- 0.01, log beta(II)(102) = 11.39 +/- 0.01. PMID- 17117221 TI - Cluster increments for macropolyhedral boranes. AB - Cluster increments derived for individual cluster fragments reproduce the DFT computed relative stabilities of macropolyhedral boranes usually within +/-6 kcal mol(-1). A simple summation procedure helps to select the best partner for a given cluster fragment in order to construct the thermodynamically most stable macropolyhedral borane. Cluster increments are considerably smaller for nido cluster fragments with an even number of vertexes than for odd nido-cluster fragments pointing towards high thermodynamic stability of macropolyhedral boranes with even numbered nido-units. PMID- 17117222 TI - Evidence for intramolecular aromatic-ring stacking in the physiological pH range of the monodeprotonated xanthine residue in mixed-ligand complexes containing xanthosinate 5'-monophosphate (XMP). AB - The stability constants of the mixed-ligand complexes formed between Cu(Arm)2+ [Arm = 2,2'-bipyridine (Bpy) or 1,10-phenanthroline (Phen)], and the di- or trianion of xanthosine 5'-monophosphoric acid [= XMP(2-) or (XMP - H)(3-)] were determined by potentiometric pH titration in aqueous solution (25 degrees C; I = 0.1 M, NaNO3). Those for the monoanion, i.e., the Cu(Arm)(H;XMP)+ complexes, could only be estimated; for these species it is concluded that the metal ion is overwhelmingly bound at N7 and the proton resides at the phosphate group. Similarly, in the Cu(Arm)(XMP)+/- [= Cu(Arm)(X - H.MP.H)+/-] complexes Cu(Arm)2+ is also at N7 but the xanthine residue has lost a proton whereas the phosphate group still carries one, i.e., stacking plays, if at all, only a very minor role, yet, the N7-bound Cu(Arm)2+ appears to form an outer-sphere macrochelate with P(O)2(OH)-, its formation degree being about 60%. All this is different in the Cu(Arm)(XMP - H)- complexes, which are formed by the (XMP - H)(3-) species, that occur at the physiological pH of 7.5 and for which previously evidence has been provided that in a tautomeric equilibrium the xanthine moiety loses a proton either from (N1)H or (N3)H. In Cu(Arm)(XMP - H)- the phosphate group is the primary binding site for Cu(Arm)2+ and the observed increased complex stability is mainly due to intramolecular stack (st) formation between the aromatic-ring systems of Phen or Bpy and the monodeprotonated xanthine residue of (XMP - H)(3 ); e.g., the stacked Cu(Phen)(XMP - H) isomer occurs with approximately 76%. Regarding biological systems the most important result is that at physiological pH the xanthine moiety has lost a proton from the (N1)H/(N3)H sites forming (XMP H)(3-) and that its anionic xanthinate residue is able to undergo aromatic-ring stacking. PMID- 17117223 TI - Electrocatalytic reduction of oxygen at electrodes coated with a bimetallic cobalt(II)/platinum(II) porphyrin. AB - Edge plane pyrolytic graphite (EPG) electrodes coated with 5-(4-pyridyl)-10,15,20 tris(3-methoxy-4-hydroxyphenyl)porphyrin and its Pt(II) and Co(II)/Pt(II) analogs undergo an electrochemical-chemical-electrochemical (ECE) reaction when anodically scanned in 1.0 M HClO4. The new redox couple formed from this anodic conditioning of the coated electrode is dependent on the pH of the solution. Roughened EPG electrodes coated with the Co(II)/Pt(II) bimetallic porphyrin show a catalytic shift of 500 mV for the reduction of O2 when compared to the reduction of O2 at a bare EPG electrode. An additional catalytic shift of ca. 100 mV is observed for O2 reduction at an EPG electrode coated with the Co(II)/Pt(II) porphyrin which has been oxidized in 1.0 M HClO4. In addition to the added electrocatalysis a significant percentage of O2 reduced at the oxidized Co(II)/Pt(II) EPG electrode is converted to H2O as determined by rotating disk electrode measurements. PMID- 17117224 TI - Dinuclear palladium(II) compounds with bridging cyclometalated phosphines. Synthesis, crystal structure and electrochemical study. AB - The structural characterization of bis-cyclometalated palladium(II) compounds of formula Pd2[(micro-(C6X4)PPh2]2(micro-O2CR)2 [X = H, R = CH3 (3), CF3 (4), C(CH3)3 (5) and C6F5 (6); X = F, R = CH3 (7) and CF3 (8)], has confirmed its paddle wheel structure with two palladium atoms bridged by two acetates and two metalated phosphines in a head-to-tail arrangement. The Pd...Pd distances are in the range 2.6779(16)-2.7229(8) A. Under cyclic voltammetric conditions, compounds 3-6, in CH2Cl2 solution, were found to undergo a reversible oxidation peak in the range of potential values 0.84-1.25 V. A second partially-reversible oxidation is observed at more positive potentials (1.37-1.55 V). For compounds 3-5 in the presence of chlorides, the first oxidation becomes a two-electron process presumably leading to a neutral [Pd(III)-Pd(III)] species with a metal-metal bond. PMID- 17117225 TI - Therapeutic claims of African potato: Fact or fiction? PMID- 17117226 TI - Hypoglycaemic and hypotensive effects of Momordica charantia Linn (Cucurbitaceae) whole-plant aqueous extract in rats. AB - Various morphological parts (roots, stems, leaves and fruits) of Momordica charantia Linn (family: Cucurbitaceae) are used traditionally in African folk medicine to manage, control and/or treat a plethora of human ailments, including diabetes mellitus and hypertension. In order to scientifically appraise some of the folkloric, anecdotal and ethnomedical uses of M charantia, the present study was undertaken to investigate the hypoglycaemic and hypotensive effects of M charantia whole-plant aqueous extract (MCE) in rat experimental paradigms. The hypoglycaemic effect of the plant extract was examined in normal and diabetic rats, using streptozotocin (STZ)- induced diabetes mellitus models. Normotensive (normal), and hypertensive Dahl salt-sensitive rats were used to probe the hypotensive (antihypertensive) effect of the plant extract. Chlorpropamide was used as reference hypoglycaemic agent for comparison. Acute oral administrations of the plant extract caused dose-related, significant hypoglycaemia in normal (normoglycaemic) and STZ-treated, diabetic rats. Furthermore, acute intravenous administrations of MCE produced dose-dependent, significant reductions in systemic arterial blood pressure and heart rates of normal, and hypertensive Dahl salt-sensitive rats. Although the exact hypoglycaemic and hypotensive mechanisms of action of the plant extract remain speculative at the moment, it is unlikely that the herb causes hypotension in the mammalian experimental animal model used via cholinergic mechanisms, since its cardiovascular effects are resistant to atropine pretreatment. However, the findings of this experimental animal study indicate that the plant extract possesses hypoglycaemic and hypotensive properties, and therefore, lend pharmacological credence to folkloric, ethnomedical uses of the plant in the management and/or control of diabetes mellitus and hypertension in some rural African communities. PMID- 17117227 TI - Experience with adjunctive corticosteroids in managing tuberculous pericarditis. AB - OBJECTIVES: To compare the efficacy of intrapericardial corticosteroid therapy to either oral corticosteroid therapy or intrapericardial placebo in addition to closed pericardiocentesis and anti-tuberculous therapy in patients with tuberculous pericarditis. METHODS: Patients with large pericardial effusions requiring pericardiocentesis were included. A short-course anti-tuberculous regimen was initiated and patients were randomised to one of three treatment groups: 200 mg intrapericardial triamcinolone hexacetonide; oral prednisone plus intrapericardial placebo; or 5 ml intrapericardial 0.9% saline (placebo). Patients were followed up for at least one year. RESULTS: Fifty-seven patients were included in the study; 21 tested HIV positive (36.8%). Forty (70.0%) had microbiological and/or histological evidence of tuberculosis, and 17 (30.0%) had a diagnosis based on clinical and laboratory data. All patients responded well to initial pericardiocentesis. However, nine patients (16.0%) were lost to follow up. The hospitalisation duration for the steroid groups was shorter than for the placebo group. This difference was not significant. Complications were similar for all arms. CONCLUSIONS: Intrapericardial and systemic corticosteroids were well tolerated but did not improve the clinical outcome. The standard six-month regimen was effective regardless of HIV infection. The potential benefits from adjunctive corticosteroids in the management of effusive tuberculous pericarditis could not be demonstrated in this three-year study. PMID- 17117228 TI - Melatonin prevents cardioprotection induced by a multi-cycle ischaemic preconditioning protocol in the isolated perfused rat heart. AB - The powerful cardioprotective actions of melatonin, the chief secretory product of the pineal gland, have been attributed largely to its free radical-scavenging properties. Free radicals play an important role in the triggering action of ischaemic preconditioning, the phenomenon whereby exposure of the heart to one or more short episodes of ischaemia leads to protection against a subsequent long period of ischaemia. The aim of this study was, therefore, to establish whether melatonin, in view of its free radical-scavenging ability, would affect the beneficial actions of preconditioning. Isolated, perfused, working hearts were subjected to 1 x 5 minute or 3 x 5 min ischaemic preconditioning protocols, in the presence or absence of melatonin (50 microM), followed by 20 minutes global ischaemia and 30 minutes reperfusion. Use was also made of sodium nitroprusside (100 microM), a nitric oxide (NO) donor and preconditioning mimetic. Using functional recovery as the endpoint, melatonin abolished the cardioprotective effects of a multi-cycle (3 x 5 min) preconditioning protocol, while having no effect on a one-cycle (1 x 5 min) protocol or SNP (1 x 5 or 3 x 5 min) preconditioning. The results suggest that free radicals play an important role in the cardioprotection induced by a multi-cycle ischaemic preconditioning protocol and that this process could be attenuated by a potent scavenger such as melatonin. PMID- 17117229 TI - The prevalence of microalbuminuria and ECG left ventricular hypertrophy in hypertensive patients in private practices in South Africa. AB - INTRODUCTION: In South Africa a cluster of cardiovascular diseases accounts for 17% of all deaths. It is also predicted that South Africa will face an epidemic of chronic kidney disease due to obesity, type 2 diabetes and hypertension. It is important to estimate the burden of underlying cardiovascular and renal disease in South Africa. OBJECTIVES: The primary objective of the study was to establish the prevalence of left ventricular hypertrophy by ECG criteria, and micro- and macroalbuminuria in mild, moderate and severe hypertensive groups. The secondary objective was to establish the prevalence of left ventricular hypertrophy and micro- and macroalbuminuria in hypertensive patients with underlying type 2 diabetes. METHODS: Patients > or = 35 years with essential hypertension with or without type 2 diabetes were recruited from 100 general practices throughout South Africa. BP, weight, height, waist circumference and urinary albumin/creatinine ratio were measured, and an ECG was performed. The Sokolow Lyon and Cornell criteria were used for estimation of left ventricular hypertrophy. An overall prevalence rate was determined using weights from the severity category distribution inside the study population. RESULTS: One thousand and ninety-one patients were available for analysis. There were 530 (48.5%) males and 561 (51.5%) females, 691 (63.3%) whites, 162 (14.8%) blacks, 150 (13.7%) Asians and 88 (8.1%) Coloureds. Of the patients, 10.9% had newly diagnosed hypertension, 20% had type 2 diabetes, and 38.1% mild, 32.1% moderate and 30.6% severe hypertension. The prevalence of left ventricular hypertrophy in the sample weighted to the hypertensive population over the age of 35 years with access to medical aids was 18.9%. Relative to white ethnicity, black ethnicity (OR 2, p = 0.03), and relative to mild hypertension, moderate (OR 2.2, p = 0.05) and severe hypertension (OR 4.9, p < 0.0001) were independent predictors of left ventricular hypertrophy. The overall prevalence of micro- and macroalbuminuria in the weighted sample was 21.3 and 4.1%, respectively. In the diabetics the prevalence of microalbuminuria was 32.3% and macroalbuminuria 10.4%, respectively. The independent predictors of microalbuminuria or worse were severe hypertension (OR 2.9, p < 0.0001), type 2 diabetes (OR 2.5, p < 0.002), and Asian ethnic group (OR 2, p = 0.02). CONCLUSIONS: The study, based on a convenience sample of hypertensives from private practices in South Africa showed that the prevalence of left ventricular hypertrophy and microalbuminuria or worse was 18.9 and 25.4%, respectively. PMID- 17117230 TI - Prophylaxis of venous thrombosis in medical patients: A real-world perspective. AB - BACKGROUND: Although low-molecular weight heparins (LMWH) have been proven to be efficacious for prophylaxis of venous thrombo-embolism (VTE) in non-surgical patients, their use and safety outside the setting of a clinical trial has not been investigated. OBJECTIVE: The objective of this survey was to determine the efficacy and safety of LMWH (enoxaparin) in the prevention of VTE in a study population comprising general medical patients. METHODS: The study involved an open-label, non-controlled, multicentre survey of any patient confined to bed due to medical illness, where the physician had made an independent decision to prescribe LMWH as prophylaxis for VTE. The demographic information and risk factors for venous thrombosis and dose of enoxaparin were recorded. Patients were assessed for clinical evidence of VTE. Only if this was present were further invasive investigations performed. Adverse events relating to the use of LMWH were recorded. RESULTS: Four hundred and seventy-one patients were enrolled from 53 centres. Five per cent of the patients were treated for up to and including three days, 4.4% for four days or less, 49.6% for six days or less, and a further 45.5 % for seven to 21 days. The most frequently prescribed dose was enoxaparin 40 mg once daily (86%). Of the enrolled patients, 28.2% had one risk factor and 69.8% had two or more risk factors for the development of VTE. The incidence of clinically suspected deep-vein thrombosis (DVT) or pulmonary embolism (PE) in this survey was three out of 457 patients at risk, ie, 0.66%. One serious adverse event occurred in an incorrectly enrolled surgical patient and 22 minor adverse events occurred that were thought by the enrolling physician to be related to the study drug. CONCLUSION: This survey was not designed to test for efficacy in DVT prevention. However the incidence of clinically identified DVT was low and there were no deaths from PE. This probably represents a considerable reduction in morbidity and mortality from that described in studies comparing LMWH with placebo. Enoxaparin may be used with confidence in a 'real-world' situation out of the confines of a study in which exclusions, inclusions and therapy are rigorously controlled. PMID- 17117231 TI - Propionibacter acnes complicating HIV-associated tuberculous pericardial effusion in Cameroon. AB - Large pericardial effusions are common in sub-Saharan Africa. They are most often caused by tuberculosis and are frequently associated with HIV infection. We report on a case of massive pericardial effusion in a 32-year-old HIV-positive male with smear-positive pulmonary tuberculosis. This was found to be colonised by Propionibacter acnes, which is normally found in anaerobic skin flora. Despite management, including appropriate antibiotic treatment based on culture and sensitivity, the patient died several weeks later. While cases of bacterial pericarditis have been described previously, this is the first report of Propionibacter isolated from a pericardial effusion. This case suggests that superinfection could have occurred iatrogenically as a result of pericardiocentesis three weeks prior to admission. We feel that proper sterile techniques, including generous use of povidone-iodine, which is readily available in resource-poor settings, may improve clinical outcomes in immunosuppressed patients undergoing invasive procedures. PMID- 17117232 TI - Off-pump myocardial revascularisation in an octogenarian patient with dextrocardia and situs inversus. AB - Dextrocardia associated with situs inversus totalis is a rare condition and there are few reports of myocardial revascularisation in such patients. An 82-year-old woman with dextrocardia and situs inversus totalis underwent successful off-pump coronary artery bypass grafting using internal mammary arteries. The operative technique was similar to that of off-pump coronary artery bypass grafting for situs solitus. However, for a right-handed surgeon the operation was easier standing on the left side of the patient. PMID- 17117233 TI - Bilateral axillary and suprascapular neuropathy after coronary artery bypass surgery. AB - Neuropathies after coronary artery bypass (CABG) and cardiac surgery are usually transient and have been described fairly frequently. Our patient presented with an acute onset of a surprisingly symmetrical bilateral axillary and suprascapular neuropathy during the immediate CABG period (which to our knowledge has not been reported on before). The patient, aged 64 years, suffered from diabetes mellitus type 2 and hypertension, and underwent a triple CABG. Electroneuronographic and electromyographic examination confirmed the presence of the bilateral axillary and suprascapular neuropathy, which showed only minimal improvement 15 months later. The surmise is that the neuropathy developed as a result of compression or overstretching of the involved nerves during the surgical procedure, associated with suboptimal positioning of the arms. The 'hands-up position', with careful sternal retraction (Cooley retractor), may appear to prevent the posterior displacement of the shoulders and plexus injuries. The diabetes mellitus may have acted as an aggravating factor. PMID- 17117234 TI - Amlodipine-based regimen to lower blood pressure, reduces risk of new-onset diabetes in hypertensives by more than a third. PMID- 17117235 TI - New clinical data shows enoxaparin reduces risk of repeat heart attacks and stroke in patients undergoing PCI. PMID- 17117236 TI - From the World Congress of Cardiology, Barcelona, 2006--Support for wider metformin usage in type 2 diabetes. PMID- 17117237 TI - Effective disease management: Today's clinical interventions to control hypercholesterolemic patients. PMID- 17117238 TI - Delaying the incubation phase from obesity to cardiovascular disease. PMID- 17117239 TI - Low awareness of impact of cardiovascular disease in low- and middle-income countries. PMID- 17117240 TI - Effective BP control, aspirin use and emphasis on post-prandial glucose control: Cornerstones of modern CV risk management. PMID- 17117241 TI - The link between erectile dysfunction and cardiovascular disease. PMID- 17117242 TI - CIBIS III trial: Bisoprolol treatment for CHF leads to 46% reduction in sudden death after one year. PMID- 17117243 TI - HIV status does not affect cardiac therapy effectiveness. PMID- 17117244 TI - Eplerenone reduces morbidity and mortality after acute MI. PMID- 17117245 TI - Genomic-based nursing care for women with Turner Syndrome: genomic-based nursing care. AB - Biologic and technologic advances generated from The Human Genome Project are having a dramatic impact on the expanding role of nurses in current health care practice. New genetic research needs to be transformed rapidly into clinical protocols with recommendations for delivering care to targeted populations. Nurses can contribute significantly, as part of an interdisciplinary approach, to translate genome-based knowledge into benefits for health care and society. In this context, we describe a clinical-genetic investigation protocol, as well nursing diagnosis, interventions and outcomes for clients with Turner Syndrome (TS) at risk for develop gonadal tumors, due the presence of a normal or abnormal Y chromosome. PMID- 17117246 TI - The nursing work process in care for healthy children at a social security institution in Mexico. AB - We aimed to analyze the nursing work process directed at care in the Nutrition, Growth and Development Surveillance and Control Program (VNCD) for children under five years at a social security institution in Mexico. The study adopted a qualitative approach and was based on the work category, on conceptions of the work process in health and on institutional analysis. We carried out direct systematic observations and interviews with a group of nurses and their assistants and with mothers who attended nursing appointments with their children. The care process was identified as procedure-centered and based on care protocols, with rapid appointments and little room for interaction between nurses and mothers. However, on some occasions, nurses were capable of producing live work as a result of their self-government, which allowed them to establish a mother/child-centered care nucleus. PMID- 17117247 TI - [The therapeutic communication during installation of endovenous therapy: the use of video-taped simulation]. AB - The use of therapeutic communication contributes to nursing care quality, mainly in procedures that cause fear and anxiety in patients, such as the installation of endovenous therapy. This study aimed to identify how nurses perceive therapeutic communication during the installation of endovenous therapy, as shown in a filmed simulation. This exploratory-descriptive and comparative study was developed in a Mexican city and involved 30 clinical nurses and 12 nursing faculty. The methodological process was: Construction and validation of textual content; Filming textual content with actors and validation; Presentation of the video to nurses and faculty and application of the semistructured questionnaire to identify therapeutic communication techniques in the simulation; perception of nurse-patient communication quality; perception of similarities between care practice and simulation; and difficulties to use these techniques in daily practice. Participants perceived the interpersonal relationship shown as "good"; they identified therapeutic communication techniques, which mostly corresponded to the description of the experience and expression of thoughts and feelings; no substantial difference was found between the perceptions of educational and care personnel. The most frequent difficulty to use communicative techniques was attributed to patients' problems to establish relations. PMID- 17117248 TI - Meanings of the nursing diagnosis implementation process for nurses at a university hospital. AB - This qualitative study aims to understand the meanings nurses at a university hospital attribute to the implementation process of the Nursing Diagnosis Classification System (DEn) as a phase in the Nursing Care System (NCS). Data were collected through interviews with eight nurses from the Medical Clinical Unit, who participated in the creation of an instrument to implement the DEn in the NCS. In their reports, the respondents expressed a positive change in their feelings, from initial discomfort and adverse perception of the change proposal. The stepwise appropriation of the process stages allowed them, besides the sharing of feelings, decisions and responsibilities for the results, to develop the belief that they would able to overcome the difficulties. PMID- 17117249 TI - Nursing phenomena identified in family planning visits with ICNP - Beta version 2. AB - This descriptive, exploratory, retrospective survey, carried out at a family planning service, aimed to identify nursing phenomena during nursing visits according to the ICNP, Beta version 2. Data were collected based on 52 records of nursing visits, realized from October 2001 to December 2002. To conduct the cross mapping process, all identified nursing phenomena were joined, organized and compared according to the ICNP's terms. Of the 51 identified nursing phenomena/diagnoses, 46 (90.2%) showed exact and partial concordance. The identified nursing phenomena can be used to assist nurses to provide care for clients in family planning services. The ICNP showed to be a comprehensive program, although some terms need to be reviewed and others enhanced. However, considering that it is an international classification applicable to several countries, the mapping process and cross-references were very satisfactory. PMID- 17117250 TI - Prenatal care: difficulties experienced by nurses. AB - This study aimed to identify the difficulties nurses experience at the start of their professional life in prenatal care activities. Data were collected through interviews with 25 nurses who accompanied prenatal care in the basic health network of Rio Branco-AC, Brazil and were grouped according to the frequency and level of difficulty they mentioned. We observed that nurses did not demonstrate difficulties in a series of important prenatal care activities at the start of their professional life. However, they reported different levels of difficulties in other activities. Furthermore, the participants pointed out difficulties in activities that require knowledge (knowing) as well as abilities (know-how). This study also indicated flaws in undergraduate formation with respect to prenatal care, involving theoretical aspects as well as exclusively practical activities. PMID- 17117251 TI - Nutritional guidance during prenatal care in public health services in Ribeirao Preto: discourse and care practice. AB - This study aimed to verify if pregnant women attended in prenatal care services at Basic Health Units in Ribeirao Preto-SP, Brazil, received nutritional guidance and if this guidance was pertinent to their nutritional status. Ninety-one pregnant women participated. The pregnant women were classified according to their nutritional condition, using a weight, height and pregnancy stage table established by the Brazilian Health Ministry's technical prenatal care manual. We found pregnant women with weight under (13.19%) and exceeding normal levels (37.36%). Independently of their nutritional condition, most of them (60.43%) declared they did not receive nutrition guidance. The mean number of prenatal visits did not influence the nutritional status. The results reveal deficiencies in the contents and quality of nutritional care. This suggests the need for care changes so as to turn discourse into practice. PMID- 17117252 TI - Self-esteem of raped women. AB - This qualitative study shows the results of workshops held with health workers and public health users (raped women), aimed at raising these women's self-esteem and creating awareness among health workers who attend them. Neuro-Linguistic Programming techniques were used to bring back life experiences, which contributed to a re-reading and to minimize causal factors of low self-esteem. Themes like repugnance, fear and the fruit of rape; image and place; death; revenge; support and solidarity; domestic violence and bad care delivery to victims were addressed during the meetings. The stories were transcribed and analyzed, preserving content fidelity. Experiences lived at home and with loved and admired people, and mainly experiences resulting from the rape were responsible for the low self-esteem. The evaluations indicated the workshops as an opportunity to reflect, to return to normal life and to reconstruct self esteem, for the raped women as well for the health workers who deliver care to them. PMID- 17117253 TI - Multidimensional tools: application of pain quality cards in children. AB - This pilot, descriptive and field exploratory study aimed to verify the representative of the quality of pain, applying the Pain Quality Cards to 50 children and teenagers hospitalized in the first half of 2004, after being approved by the Ethic Commission. Results have shown that there is no relationship between the age group and the number of positive answers. The identification of the cards was different to each group, 61.1% of the cards were identified for the pre-scholar, 77.8% for the scholar and 27.8 for the teenagers. The use of the instrument has revealed itself successful and able to evaluate, discriminate and measure the different dimensions of pain. PMID- 17117254 TI - Qualitative and quantitative aspects of pain in lateral posterior thoracotomy patients. AB - Descriptive study that proposed to compare the qualitative and quantitative behavior of the pain in lateral posterior thoracotomy patients. The sample was consisted of 18 individuals with an average age of 44 years. The instruments used were physiotherapy evaluation form, numerical pain scale and McGill questionnaire for pain. The pain on the numerical pain scale was considered moderate(5) for both sexes. The descriptors of the McGill questionnaire choosen by the patients with higher frequency were: in the sensorial component, beat4, pointed1, shock2, final and pull2; in the afetive component, tired1, bored1, punishald1 and miserable1 and in the evaluative component was flat. The characteristics of pain in the sensorial group were more evidents on male group. No significant statistical difeferences were observed between quantitative answers concerning pain between the men and women. On the qualitative aspects , was observed an predominancy of the same descriptors of pain in afetive component for both sexes. Pain intensity was categorized as moderate. No significant statistical difference were observed between the pain on the post-operatory lateral posterior thoracotomy. These data demonstrate a necessity for an analysis with a larger study group. PMID- 17117255 TI - Relation between stressors and sociodemographic and clinical characteristics of patients hospitalized at a coronary unit. AB - This descriptive study aimed to correlate the stressors of patients hospitalized at a coronary care unit and their sociodemographic and clinical characteristics. We interviewed 43 patients who were hospitalized at a large hospital. The stressors were evaluated by means of a 4-point Likert scale, which measured stress intensity for 42 possible stressors, ranging from 1 (not stressful) to 4 (very stressful). We collected data on sociodemographic and clinical characteristics and about the coronary unit. Data were analyzed through non parametrical statistics, using Mann-Whitney, Kruskal-Wallis and Spearman's correlation test. A 0.05 significance level was adopted. Greater stress was found among younger female patients who did not receive psychotherapeutic medication, including the presence of more than two pieces of equipment and no earlier hospitalization at this kind of intensive therapy unit. PMID- 17117256 TI - Nurses' understanding of standard precautions at a public hospital in Goiania - GO, Brazil. AB - Standard Precautions (SP) are effective strategies to prevent and control nosocomial infections. This study aimed to verify nurses' understanding about standard precaution measures. Data were collected through interviews, followed by content analysis in accordance with Bardin. Eighty-two nurses took part in this study, 75.6% of whom understand SP as protective measures: for professionals (11.0%); for both professionals and patients (52.4%); for patient care independently of the diagnosis (7.3%); for patients with diagnosed infection (9.8%). Other nurses indicated SP as human care (4.9%) and only as Individual Protection Equipment (IPE) (11.0%). Most participants' understanding points to favorable cognitive adaptation to the daily implementation of SP. However, reductionist and even mistaken perceptions about their range persist, which makes the social function of these measures vulnerable. PMID- 17117257 TI - Knowledge on diabetes mellitus in the self care process. AB - This descriptive study in the interior of Sao Paulo aimed to verify diabetes mellitus patients' knowledge about the disease, causes and complications, highlighting its importance in self care. Data were collected through interviews with 84 persons and analyzed through descriptive statistics. Average age was 53.3 +/- 13 years, time of disease 12.9 +/- 9 years and 58% of the participants did not finish basic education. Only 28.6% of the participants gave correct answers to "what is diabetes" and "what are its causes"; 71% were diagnosed without presenting classic symptoms and 64% had already been hospitalized due to an acute or chronic complication. We indicated aspects that turn the learning process more difficult; little knowledge about the disease, its causes and symptoms, thus affecting the prevention and early diagnosis and entailing predisposition towards complications. Furthermore, the interference of biopsychosocial factors in the self care process is highlighted. PMID- 17117258 TI - Evaluation of cleaning and disinfection performance of automatic washer disinfectors machines in programs presenting different cycle times and temperatures. AB - Thermal washer-disinfectors represent a technology that brought about great advantages such as, establishment of protocols, standard operating procedures, reduction in occupational risk of a biological and environmental nature. The efficacy of the cleaning and disinfection obtained by automatic washer disinfectors machines in running programs with different times and temperatures determined by the different official agencies was validated according to recommendations from ISO Standards 15883-1/1999 and HTM2030 (NHS Estates, 1997) for the determining of the Minimum Lethality and DAL both theoretically and through the use with thermocouples. In order to determine the cleaning efficacy, the Soil Test, Biotrace Pro-tect and the Protein Test Kit were used. The procedure to verify the CFU count of viable microorganisms was performed before and after the thermal disinfection. This article shows that the results are in compliance with the ISO and HTM Standards. The validation steps confirmed the high efficacy level of the Medical Washer-Disinfectors. This protocol enabled the evaluation of the procedure based on evidence supported by scientific research, aiming at the support of the Supply Center multi-professional personnel with information and the possibility of developing further research. PMID- 17117260 TI - Critical incidents in the teaching-learning process of a nursing course through the perception of students and faculty. AB - The teaching-learning process is complex and leaves many question marks, mainly when one thinks about quality. Therefore, this study aims at identifying factors that interfere positively or negatively in the teaching-learning process, through the perspective of students and faculty of the Nursing course at the Universidade Norte do Parana- UNOPAR. This descriptive study with a qualitative approach was carried out through the critical incidents technique. Thirty-six faculty and 140 students participated. Data analysis revealed that the students mentioned 435 critical incidents related to the category faculty behavior, being 317 negative references and 118 positive. According to the faculty members' reports, the category interaction with the group produced 58 references, being 10 positive and 48 negative. An adequate teaching-learning process requires good faculty-student and student-student relationships, favoring good interaction and efficient learning. PMID- 17117261 TI - Training program on microbiological test collection material methods at a teaching hospital: investment and result assessment. AB - This study aimed at evaluating the results, direct costs and investment of a training program on microbiological test material collection at a teaching hospital. Test collections that did not follow the established criteria (failure) were considered as the result measure. Variable and absorption costing were used to calculate direct costs and investments, respectively. Of the 11,893 collected materials, failures were evidenced in 59 (0.5%). Direct cost corresponded to R$ 154.10 and R$ 2,431.29 was invested in training. These findings revealed that the evidenced number of anomalies (failures) represented a low percentage in relation to the total collected material for microbiological exams. Therefore, this should not be considered a critical point that justifies the continuity of the training and, consequently, the investment. PMID- 17117262 TI - Nursing education: seeking critical-reflexive education and professional competencies. AB - The study describes changes that are noted in students during training and which contribute to define a professional profile. We carried out a descriptive exploratory study with a qualitative approach, based on dialectics as a theoretical-methodological framework. The data was obtained from documented analysis and through focal groups with teachers, students and service nurses. The results show the student's position as an active subject in the teaching-learning process, through a movement of transformation of academic structures. A correlation was found between the movement that seeks students' greater political, active and critical participation as a way of determining and guiding the profile of the generalist nurse and greater social insertion. We conclude that, despite efforts, training guidelines and the definition of the professional profile in the study settings is directed at the demands of the labor market, and that competency-based training is still incipient. PMID- 17117263 TI - Scientific nursing production from the perspective of social representation. Brazil, 1975-2001. AB - This study aimed to: survey the number of nursing theses and dissertations produced between 1975 and 2001 with a social representation approach (RS); characterize the products with regard to the institution of origin, country region, study subjects and representation objects; to group in to theme categories and research lines in nursing. The data were collected from the Center of Studies and Research in Nursing at the Brazilian Nursing Association. The result evidenced 131 studies with RS support, originated in 1990, with higher production in the year 2000, mainly in the South East (83.1%). It registered 145 subjects; the most common objects are "disease", "health-disease process", "nurses' professional practice" and "care delivery". In the thematic categorization, the care area joins the greatest quantity of studies. In short, nursing has been seeking to appropriate itself of this theoretical-methodological framework as the basis and guide for part of its knowledge production. PMID- 17117264 TI - (Handicapped) caregiver: the social representations of family members about the caregiving process. AB - This study aimed to analyze what looking after physically disabled persons with spinal cord injury by trauma means to their caregivers and family members. The analysis of the testimony of eight caregivers, obtained in open interviews, which was methodologically based on the Social Representations Theory (SR), pointed out two main routes: coping with the suffering process in care practice and the troubled waters that permeate this suffering process. These two routes, characterized as SR Central Core and Peripheral System, respectively, consisted of themes like the way of looking at impairment, affectivity, religiosity, social economical changes and (lack of) technical and institutional support. The results show a handicapped caregiver dedicated to look after someone who is physically disabled, considered incapacitated, and who leads his or her chores with distress and privations, based on guilt and religiosity, supported by ambiguous affection and affected by deteriorating social-economical changes and (lack of) technical and institutional support to practice an activity that implies so many peculiarities. The transformation alternatives of these caregivers' daily life principally lead to a symbiosis of disability with the patient - to live for the physically disabled - or yet, for a few, a sketch to restart personal life projects - to live with the physically disabled. PMID- 17117265 TI - HIV positive mothers' beliefs about mother-to-child transmission. AB - Beliefs can influence health behavior. This qualitative study aimed to understand the beliefs that influence HIV positive mothers' behaviors towards prevention methods against mother-to-child transmission. Fourteen women were interviewed. Our research was based on the theoretical Health Belief Model, formed by the following dimensions: perceived susceptibility, perceived severity, perceived benefits and perceived obstacles. Data analysis showed reflections that evidence the paradox in the AIDS epidemic: knowledge does not change behavior; gender relations; fear of death; fear of stigma; financial problems; disbelief in the virus' existence. Identifying beliefs and understanding how to influence the conduction of the health problem can help services to promote patients' adherence. PMID- 17117266 TI - Review of scientific literature on the use of group activities in nursing work in Brazil: 1980 to 2003. AB - Among the many and diverse activities carried out by nurses, many are organized within the context of a group approach. The realization that there were no systematized studies in Brazil, capable of providing parameters that would help to understand how nurses use this resource, motivated the authors to develop this study, whose objective was to review scientific literature on the use of group activity, as it relates to the work carried out by nurses, between 1980 and 2003. This was a descriptive/exploratory research developed through a bibliographic review in Brazilian journals, on the basis of a specific protocol. We were able to identify 151 articles that were reviewed in terms of the expansion of this knowledge and the different situations in which a group is used by nurses. The qualitative analysis done on pre-established categories enabled the authors to verify that nurses have been using this technique while delivering care, producing knowledge and developing human resources. This investigation allowed us to outline a panoramic view of this literature and to develop a catalogue, identifying important aspects in the construction of this knowledge by Brazilian nursing personnel. PMID- 17117267 TI - Social representations about aids in people over 50, infected by HIV. AB - This study analyzes representations about AIDS in nine persons older than 50, infected by HIV. The analysis revealed representations: 'AIDS is a constant death threat'. These were categorized and named by means of the interviewers' statements: 'No doctor thinks, at first, that we can have aids; AIDS isn't cancer; to be old and have AIDS is to be discriminated twice. The results show the importance of integral care delivery by health services to decrease the psychosocial suffering of these people. PMID- 17117268 TI - Mental health and work: meanings and limits of theoretical models. AB - This text is a revision of the most important theoretical approaches that fundament the studies about Mental Health and Work, pointing to the basic concepts, categories of analyses and respective limits. The objective is to underline some conceptions about the theme, in the attempt to contribute with a way to face the problems, questions and difficulties imposed by the construction of this object of study. Thus, the text is based on the critical review of theoretical paradigms that predominate in studies about Mental Health and Work. Finally, it indicates some important theoretical questions about the relation between mental health and work. PMID- 17117269 TI - Data collection in clinical-qualitative research: use of non-directed interviews with open-ended questions by health professionals. AB - Non-directed interviews constitute the main data collection instrument in qualitative health research. Studies in which this is evident are well documented in international literature. For health professionals, knowing what people feel and imagine makes it possible to develop a more adequate clinician-patient relationship. It is indispensable to know what the life phenomena mean for individuals, because the meanings have a structuring function. People organize their lives around the meaning they attribute to situations or object. This is also relevant to their health care. From research conducted at the Laboratory of Clinical-Qualitative Research, State University of Campinas, Campinas (Sao Paulo), Brazil, the authors address, in this article, the following matters: characterization of non-directed interviews, directiveness of interviews, approach techniques, observation of non-verbal and paraverbal manifestations, registry techniques / speech transcription, and validity/reliability of non directed interviews. This is useful for people interested in research at graduate and undergraduate level. PMID- 17117270 TI - Historical and social aspects of halitosis. AB - Buccal odors have always been a factor of concern for society. This study aims to investigate the historical and social base of halitosis, through systematized research in the database BVS (biblioteca virtual em saude - virtual library in health) and also in books. Lack of knowledge on how to prevent halitosis allows for its occurrence, limiting quality of life. As social relationships are one of the pillars of the quality of life concept, halitosis needs to be considered a factor of negative interference. Education in health should be accomplished with a view to a dynamic balance, involving human beings' physical and psychological aspects, as well as their social interactions, so that individuals do not become jigsaw puzzles of sick parts. PMID- 17117271 TI - [Effect of Passiflora edulis (passion fruit) extract on rats' bladder wound healing: morphological study]. AB - PURPOSE: To evaluate the effects of hydroalcoholic extract of Passiflora edulis leaves in the healing of urinary bladder in rats from histological aspects. METHODS: Forty Wistar male rats were submitted to a longitudinal incision of the bladder followed by a stetching in only one level. After this common procedure, animals were divided at random two groups: Passiflora and Control. In the Passiflora group the only dosage used was administered by intraperitoneal injection of hydroalcoholic extract of Passiflora edulis leaves while in the Control group distilled water was injected. Each subgroup was then divided in two subgroups according to the death of these animals: Control, three and seven days, Passiflora, three and seven days. After the death of these animals, an inventory of the abdominal cavity was performed and the bladder was removed. A comparative analysis was done between the two groups with microscopic evaluation of the healing. There was less acute inflammation (p=0.008), greater collagenous formation (p=0.001) and greater capillary neo-formation (p=0.000) in the third day Passiflora subgroup when compared to the Control subgroup of the third day. RESULTS: There was less acute inflammation (p=0.001), greater fibroblastic proliferation (p=0.011) and greater collagenous formation (p=0.001) in the Passiflora subgroup of seventh day when compared with the Control seventh day subgroup. CONCLUSION: The use of Passiflora edulis leaves extract resulted in less acute inflammation, greater fibroblastic proliferation, collagenous formation and capillary neo-formation on rats' bladder wound healing. PMID- 17117272 TI - [Passiflora edulis extract and the healing of abdominal wall of rats: morphological and tensiometric study]. AB - INTRODUCTION: The Brazilian popular habit of using plants to treat several health conditions is ancient. Passion fruit (Passiflora edulis) is widely used to treat, usually in an empiric basis, a variety of medical conditions. Anti-inflammatory activity of Passiflora edulis extract, similar to non-steroidal anti-inflammatory drugs (NSAID's), has been described. PURPOSE: To evaluate the effect of Passiflora edulis hydroalcoholic extract on the healing of midline abdominal incisions in rats by morphological and tensiometric methods. METHODS: Forty male Wistar rats were randomly allocated into two groups to either receive Passiflora edulis extract (study group, P) or saline (control group, C) intraperitoneally, in a single isovolumetric dose, after a standardized ventral midline laparotomy had been performed. The twenty rats of the control group as well as those twenty of the study group were divided into subgroups according to the time of sacrifice, either the 3rd PO day (P3, C3) or the 7th PO day (P7, C7). On day three and on day seven after surgery, the rats were sacrificed and the wound area was excised by a standardized protocol. The healing process of the specimens was evaluated macroscopically and histologically. The tensile strength was evaluated by a constant speed computerized tensiometer to determine the breaking strength and the deformation of the healing incision. RESULTS: The macroscopic examination did not show significant differences between study and control groups. Histologically, the C3 vs. P3 comparison showed the following differences: for the variables acute inflammation (p=0.045 in favor of C3), collagenization and capillary neoformation: p=0.001 e 0.001, respectively in favor of P3. Similarly, the C7 vs. P7 comparison showed the following differences for the variables acute inflammation (p=0.002 in favor of C7), chronic inflammation and capillary neoformation: p= 0.006 e 0.001, respectively in favor of P7. Tensiometrically, maximal breaking strength (C(max)) on day seven of the study group was higher when compared to control group, (6.91 +/- 1.36 vs. 5.05 +/- 1.63, p=0.013). Maximal deformation strength (D(max)) on day seven of the study group was higher when compared to control group (36.49 +/- 4.61 vs. 26.19 +/-5.74, p=0.001). CONCLUSION: Passiflora edulis extract enhances the healing of midline abdominal incisions in rats, especially the histological and tensiometric aspects. PMID- 17117273 TI - [Evaluation of hydroalcoholic extract of Aroeira (Shinus Terebinthifolius Raddi) in the healing process of wound skin in rats]. AB - PURPOSE: To evaluate the healing effect of the topic administration of the hydroalcoholic extract of aroeira in open wounds in the dorsocostal region of rats. METHODS: Sixty Wistar rats, males, were used. In all animals one skin fragment was removed, with two centimeters in diameter. Inhalatory anesthesia with ethyl ether was done, The rats were divided in two groups of 30 animals: aroeira group, submitted to an topic application of the hydroalcoholic extract of the plant, and control group, with the same process but with 0.9% saline solution. Each group was divided in three sub-groups of 10 animals according to the period of observation: 7, 14 and 21 days. The areas of the injuries were analyzed by the macroscopic aspect and digital planimetry, and the resected segments of the wounds were studied by optic microscopy stained with hematoxilin eosine and trichromium of Masson. RESULTS: The macroscopic findings showed early complete reepitelization in the control group by the 14th day. According to the digital planimetry the average wound areas of the rats from the control group (0.5278 cm(2)) had been smaller than those from the wounds of the rats of the aroeira group (0.6897 cm(2)), with statistical significance to the 14th day (p=0.036). The histologic study showed statistical difference (p=0.023) regarding the mononuclear cells in 14th day of evaluation, with larger number in the aroeira group, without significant differences regarding other parameters in all the remaining days. CONCLUSION: The topic use of the hydroalcoholic extract of aroeira delayed the reepitelization of the skin wounds in rats. PMID- 17117274 TI - [Healing process in bladder suture of rats with and without the administration of intraperitoneal Jatropha gossypiifolia L. gross extract]. AB - INTRODUCTION: The suture of tissues and their healing process is one of the basic fundamentals of surgery, and the research on substances which could improve the process is an ongoing challenge. The use of plants has been tested but till nowadays without scientific demonstration. PURPOSE: To compare the macroscopic and histological alterations done by the use of intraperitoneal Jatropha gossypiifolia L. gross extract, in the healing process of sutures performed on the bladder of rats. METHODS: Forty adult, male, Wistar rats were distributed into two animal groups. One centimeter longitudinal incision on the bladder ventral wall, and single plane synthesis with separated polyglactine 910 5-0 (Ethicon) stitches was done in all the animals. In the control group an intraperitoneal cavity instillation of distilled water at a ratio of 1 ml/kg/weight, and a 1 ml/kg/weight of Jatropha gossypiifolia L. extract for the Jatropha group was done representing 200 mg of the substance. Each group was subdivided into two with 10 animals in each, being submitted to euthanasia on 3rd and 7th post-operative days. Comparative histological, macroscopic and statistical analysis were undertaken between the subgroups. RESULTS: Statistical significant difference was observed in the acute inflammation changes, vascular neoformation and bonding on day 3; the first one was greater in the control group, and the latter two greater in the Jatropha group; acute inflammation variables and fibroblastic proliferation presented to be more intense on day 7, with statistical significance favoring the control group. CONCLUSION: No favorable healing effect was observed with the administration of single intraperitoneal dose of Jatropha gossypiifolia L. gross extract on the sutures of bladder in rats. PMID- 17117275 TI - [Healing of colonic anastomosis with the use of extract aqueous of Orbignya phalerata (Babassu) in rats]. AB - INTRODUCTION: Healing is a highly complex process with different phases. Because some medicinal plants have antiinflammatory properties, they could influence the wound healing process. There are many species of medicinal plants used for research, among them the Orbignya phalerata (Babassu). PURPOSE: To analyze, comparatively, the tensiometric and histologic alterations in the healing process of colon anastomosis with the use of aqueous extract of the babacu mesocarp. METHODS: Forty Wistar male rats were used, divided in two groups of 20 animals named control and experimental group. Each group was further divided in two sub groups of 10 animals and evaluated on the third and seventh post-operative days. Colotomy and one suture line coloraphy was performed in all animals. The control group received an intraperitoneal single dose of isotonic saline solution at the time of the surgery. The animals from the experimental group received babassu aqueous extract dose with 50mg/kg/weight, concentration of 25 mg/ml dose, with the same administration method. The following parameters were evaluated: a) macroscopic appearance of abdominal wall and cavity; b) tensiometric evaluation of anastomosis (bursting pressure); c) histologic characteristics of anastomosis. RESULTS: The macroscopic examination showed the presence of adherences in all animals with statistical significance in the ones from the experimental group at third day when compared to the control group. No difference was observed in the seventh-day group. No fistulae, abscesses and/or hemorrhage were observed. In regard to the tensiometric variation, the bursting pressure for the control and experimental groups, both for the third day (25.4 mmHg - 14.8 mmHg) and at seventh (183 mmHg - 175 mmHg), were similar with no significant difference. Histological analysis for the third day showed significant difference in the experimental group in relation to the controls, in regard the presence ofto mononuclear ones whereas at seventh day there was significant statistical difference in all variables from the experimental group compared to the control one. CONCLUSION: A favorable effect with administration of the aqueous extract of the babacu mesocarp was seen in histological parameters of the healing process of colon anastomosis however there was no difference in the tensiometric evaluation between the control and experimental group. PMID- 17117276 TI - [Evaluation of aroeira (Schinus terebinthifolius Raddi) extract on the healing process of gastroraphy in rats]. AB - PURPOSE: To evaluate the healing process of gastric suture in rats using hydroalcoholic aroeira (Schinus terebinthifolius Raddi) extract. METHODS: Forty adult male rats, divided into two groups of 20 animals were operated and named as follows: aroeira group (Ga), and the control group (Gc). Each group was divided into two subgroups (SG) of 10 animals (SGa and SGc) according to the time of provoked death (three and seven days). The same surgical procedure was performed in all animals consisting in incision and simple suture of the stomach (Prolene(R) 6-0). The only difference was on the type of medical treatment. The aroeira group received a single 100 mg/kg of aroeira extract in an intraperitoneal dose and the animals from the control group received the same quantity in milliliters (ml) of the isotonic saline solution. The evaluated parameters were: macroscopic alterations, microscopic healing process and toleration to atmospheric air insufflation. RESULTS: All animals had good healing process of abdominal wall with no clinical evidence of infection, dehiscence, abscesses and peritonitis. Both groups presented adherences to gastric suture line area with surrounding organs, mainly the liver, lower intestines and the abdominal wall. Microscopic analysis showed only chronic inflammation significant difference between the aroeira and control groups on the third day of observation. Resistance tests did not present significant statistical differences in the studied groups. CONCLUSION: The use of aroeira (Schinus terebinthifolius Raddi) hydro-alcoholic extract did not alter the stomach healing process, considered on macroscopic, tensiometric and microscopic assessment. PMID- 17117277 TI - [Evaluation of the aroreira (Schinus terebinthifolius Raddi) in the healing process of surgical incision in the bladder of rats]. AB - PURPOSE: To evaluate the hydroalcoholic aroeira extract in the healing process of surgical incisions in the bladder of rats in an experimental comparative study. METHODS: Forty male Wistar rats were used. All of them underwent a median abdominal incision, with an 1cm cystotomy, followed by one plain suture with separated stitches of poliglactine 910 5-0. The animals were then divided into two groups of 20 rats each, and named the aroeira group (GA) and the control group (GC). In the first one, 80% hydro-alcoholic plant extract was injected in the peritoneal cavity, in a one dose of 100mg/kg. The second group, a 0.9% saline solution was injected in place of aroeira extract. Each group was divided into two subgroups (SGA and SGC) of 10 animals. According to their scheduled death, they were named subgroups SGA3 and SGC3 when killed in the 3th day after surgery and subgroups SGA7 and SGC7 in the 7th day. The abdominal cavity and the bladder suture were macroscopically evaluated. RESULTS: The microscopic analysis of the surgical incision in the bladder was performed by means of the hematoxilin-eosine stain and the trichrome of Masson. The macroscopic analysis showed an infection in the surgical incision in three rats of the SGC group and in one of the SGA, and peritoneal adhesion in the 26 rats belonging nine to SGC and 17 in the SGA. The microscopic evaluation revealed a more severe acute inflammation process in the SGC on the 3th (p=0.045) and in the 7th (0=0.002) days. In the SGA, a more significant collagenization (p=0.001) could be seen, as far as the evidence of vascular neoformation (p=0.002) on the 3rd day. Chronic inflammatory reaction (p=0.006) and a more intense vascular neoformation (p=0.001) were observed in the 7th day. CONCLUSION: The hydroalcoholic aroeira extract had a favorable effect in the healing process of cystotomies done in rats. PMID- 17117278 TI - [Extract of Passiflora edulis in the healing process of gastric sutures in rats: a morphological and tensiometric study]. AB - INTRODUCTION: Many substances of vegetable origin have been used since the beginning of civilization with the purpose of improving the healing process. Among them, dry leaves extract from Passiflora edulis have been shown to have an anti-inflammatory effect in rats. PURPOSE: To analyze the effect of dry leaves extract from Passiflora edulis in the healing of gastric sutures in rats. METHODS: Forty male adult Wistar rats were divided into two groups of 20 rats, called Passiflora group (GP) and Control group (GC) which were divided into two groups of 10 according to moment of death, on day 3 or day 7 after the operation. All animals were submitted to a midline incision and a gastrotomy was performed on the anterior wall of the stomach followed by gastric suture with polypropylene 6.0 using four stitches on a single layer. Rats from GP were given, before closure of the abdominal wall, a solution of Passiflora edulis extract, 250 mg/kg/weight, while rats from the GC were given an isovolumetric isotonic saline solution. Macroscopic evaluation included the adhesion index proposed by Knightly. Bursting pressure was measure by an electronic device. Microscopic analysis was performed including inflammation parameters. RESULTS: All animals presented adequate healing of the abdominal wall with no clinical signs of infections or dehiscence. The adherence index was similar in both groups both on day 3 (p=0.734) and on day 7 (p=1.000). The gastric sutures presented leak with smaller insufflation pressure on the 3rd P.O. day in both groups as compared to the 3rd P.O. day. There was no significant difference of bursting pressure among the subgroups on the 3rd P.O. day (GC3=41.1 +/- 22.1 mmHg versus GP3=59.2 +/- 20.4 mmHg; p=0.074). On the 7th P.O. day, there was an increased mean bursting pressure in both groups, but there was no statistically significant difference between the two groups (p=0.850). Histologic parameters were similar in both groups, on P.O. days 3 and day 7, except for the fibroblastic proliferation, which was greater on the 7th day in GP (p=0.002). CONCLUSION: The intraperitoneal use of Passiflora edulis extract influences favorably the healing of gastric sutures in rats because of the increase in the fibroblastic proliferation on the 7th P.O. day. PMID- 17117279 TI - [Phytotherapeutic evaluation of Jatropha gossypiifolia L. on rats ventral abdominal wall wound healing]. AB - INTRODUCTION: The Jatropha gossypiifolia L., which is used in popular medicine is considered to have good diuretic effect in hypertension and is also used as a laxative drug. It seems to have a healing effect, although not proved till now. PURPOSE: To evaluate the influence of intraperitoneal administration of Jatropha Gossypiifolia L., in suture healing of ventral abdominal wall of rats, through tensiometric measurement, macro and microscopic aspect of post-operative period. METHODS: Forty Wistar male rats were allocated in two groups of 20 animals. After the incision and exposure of abdominal cavity 1 ml/kg/weight of 0,9% sodium chloride solution was injected in control group, and in the other one the injection was of 1 ml/kg/weight of a gross ethanol extract of Jatropha gossypiifolia L. The suture of the abdominal wall was than performed with polypropylene separated stitches. The animals were followed-up and killed in the third and seventh days. The ventral abdominal wall was macroscopically analyzed, the resistance strength to strain was measured and it was also studied the histological aspects. RESULTS: On macroscopic examination more intense adhesion was found on the group of Jatropha in both third and seventh post-operative days. The strain evaluation was meanly greater on Jatropha group also in third and seventh days. CONCLUSION: The histological comparative analysis between the different groups showed that the acute inflammatory process was meanly greater for the Jatropha group in third and seventh post-operative days. The vascular neoformation was significantly greater in third post-operative day of Jathopha group; the other histological parameters were just alike. The intraperitoneal injection of Jatropha extract did not have any significant improvement for the wound healing on ventral abdominal wall on the evaluated animals in this study, no matter if analyzed at the third or seventh post-operative days. PMID- 17117280 TI - [Topic use of aqueous extract of Orbignya phalerata (babassu) in rats: analysis of its healing effect]. AB - INTRODUCTION: The mesocarp of Babassu (Orbignya phalerata) has been used in experimental studies trying to check its pro-inflammatory effect. PURPOSE: To analyse comparatively the histological changes made by the water extract of Babassu in skin surgical wounds. METHODS: Sixty Wistar adult male rats were used. The experimental procedure was a circle skin incision of 2 cm in diameter made with a metal punch. After this procedure done in all animals, they were randomized in two groups of 30. In the control group, the phytotherapeutic agent wasn't used but saline solution. In the experimental group, the water extract of Orbignya phalerata was used in the cut sites. All animals were followed and killed after seven, 14 and 21 days. Comparative histological analysis was made among the groups. RESULTS: In the microscopic view, a significant reepitelization effect on the healing process of the experimental group in relation to the control group in the 7th and 14th days, was recognized. CONCLUSION: The use of the mesocarp of Orbignya phalerata in skin surgical wounds contributed positively in the healing process in rats. PMID- 17117282 TI - C'est si bone. PMID- 17117283 TI - Bone quality: what is it and how is it measured? AB - Bone quality describes aspects of bone composition and structure that contribute to bone strength independently of bone mineral density. These include bone turnover, microarchitecture, mineralisation, microdamage and the composition of bone matrix and mineral. New techniques to assess these components of bone quality are being developed and should produce important insights into determinants of fracture risk in untreated and treated disease. PMID- 17117284 TI - Bone density testing in clinical practice. AB - The diagnosis of osteoporosis and monitoring of treatment is a challenge for physicians due to the large number of available tests and complexities of interpretation. Bone mineral density (BMD) testing is a non-invasive measurement to assess skeletal health. The "gold-standard" technology for diagnosis and monitoring is dual-energy X-ray absorptiometry (DXA) of the spine, hip, or forearm. Fracture risk can be predicted using DXA and other technologies at many skeletal sites. Despite guidelines for selecting patients for BMD testing and identifying those most likely to benefit from treatment, many patients are not being tested or receiving therapy. Even patients with very high risk of fracture, such as those on long-term glucocorticoid therapy or with prevalent fragility fractures, are often not managed appropriately. The optimal testing strategy varies according to local availability and affordability of BMD testing. The role of BMD testing to monitor therapy is still being defined, and interpretation of serial studies requires special attention to instrument calibration, acquisition technique, analysis, and precision assessment. BMD is usually reported as a T score, the standard deviation variance of the patient's BMD compared to a normal young-adult reference population. BMD in postmenopausal women is classified as normal, osteopenia, or osteoporosis according to criteria established by the World Health Organization. Standardized methodologies are being developed to establish cost-effective intervention thresholds for pharmacological therapy based on T-score combined with clinical risk factors for fracture. PMID- 17117285 TI - Peripheral bone densitometry: Clinical applications. AB - Technologies for the measurement of bone mineral density and other parameters of bone strength at peripheral skeletal sites have been studied since the 1960s. Single-energy Photon Absorptiometry (SPA), Radiographic Absorptiometry (RA), Radiogrametry (RG), Single-energy X-ray Absorptiometry (SXA), Peripheral Dual energy X-ray Absorptiometry (pDXA), and Quantitative Ultrasonometry (QUS) have been successively evaluated. These technologies and their clinical applications are discussed in this article. The available scientific evidence supports the clinical use of these technologies at peripheral skeletal for assessment of fracture risk. Peripheral measurements other than the 33% (one-third) radius by DXA cannot be used to diagnose osteoporosis according to current standards. Peripheral skeletal sites are not clinically useful for monitoring changes in BMD with natural evolution of the disease and its treatment. Peripheral BMD measurement can theoretically be used to screen patients for selection to central DXA testing, although device-specific cut-points should be developed before this is implemented. When central DXA testing is not available, peripheral BMD testing may be considered to identify individuals who might benefit from pharmacological intervention. PMID- 17117286 TI - Clinical application of biochemical markers of bone turnover. AB - With the ageing population in most countries, disorders of bone and mineral metabolism are becoming increasingly relevant to every day clinical practice. Consequently, the interest in, and the need for effective measures to be used in the screening, diagnosis and follow-up of such pathologies have markedly grown. Together with clinical and imaging techniques, biochemical tests play an important role in the assessment and differential diagnosis of metabolic bone disease. In recent years, the isolation and characterisation of cellular and extracellular components of the skeletal matrix have resulted in the development of molecular markers that are considered to reflect either bone formation or bone resorption. These biochemical indices are non-invasive, comparatively inexpensive and, when applied and interpreted correctly, helpful tools in the diagnostic and therapeutic assessment of metabolic bone disease. This review provides an overview of the current evidence regarding the clinical use of biochemical markers of bone remodelling in bone disease, with an emphasis on osteoporosis. PMID- 17117288 TI - The calcium-sensing receptor and related diseases. AB - The calcium-sensing receptor (CASR) adjusts the extracellular calcium set point regulating PTH secretion and renal calcium excretion. The receptor is expressed in several tissues and is also involved in other cellular functions such as proliferation, differentiation and other hormonal secretion. High extracellular calcium levels activate the receptor resulting in modulation of several signaling pathways depending on the target tissues. Mutations in the CASR gene can result in gain or loss of receptor function. Gain of function mutations are associated to Autossomal dominant hypocalcemia and Bartter syndrome type V, while loss of function mutations are associated to Familial hypocalciuric hypercalcemia and Neonatal severe hyperparathyroidism. More than one hundred mutations were described in this gene. In addition to calcium, the receptor also interacts with several ions and polyamines. The CASR is a potential therapeutic target to treatment of diseases including hyperparathyroidism and osteoporosis, since its interaction with pharmacological compounds results in modulation of PTH secretion. PMID- 17117287 TI - Evolution of PTH assays. AB - PTH metabolism is complex and the circulating forms include the intact 1-84 molecule as well as several carboxyl-terminal fragments. The first generation of PTH assays included several types of competitive assays, with specificities that spanned carboxyl, mid-region and amino-terminal portions of the molecule. The limitations of these assays and the methodological evolution led to the description of 2nd generation non-competitive immunometric assays for PTH in the late 80's, based on the recognition of the PTH molecule by two different antibodies, one directed against de amino-terminal and other against the carboxyl terminal segments. The observation that in some circumstances "long" carboxyl terminal segments were also measured by 2nd generation assays led to the development of 3rd generation assays based on amino-terminal specific antibodies that are specific for the first amino acids, measuring only the molecular forms that activate PTH1R. The practical and cost-benefit advantages of these assays are still debatable. The recent observation that carboxyl-terminal fragments of PTH have biological activity via a distinct receptor than PTH1R, points to the future need of more than one assay in order to evaluate parathyroid hormone function. PMID- 17117289 TI - Vitamin D deficiency: A global perspective. AB - Vitamin D is essential for the maintenance of good health. Its sources can be skin production and diet intake. Most humans depend on sunlight exposure (UVB 290 315 nm) to satisfy their requirements for vitamin D. Solar ultraviolet B photons are absorbed by the skin, leading to transformation of 7-dehydrocholesterol into vitamin D3 (cholecalciferol). Season, latitude, time of day, skin pigmentation, aging, sunscreen use, all influence the cutaneous production of vitamin D3. Vitamin D deficiency not only causes rickets among children but also precipitates and exacerbates osteoporosis among adults and causes the painful bone disease osteomalacia. Vitamin D deficiency has been associated with increased risk for other morbidities such as cardiovascular disease, type 1 and type 2 diabetes mellitus and cancer, especially of the colon and prostate. The prevalence of hypovitaminosis D is considerable even in low latitudes and should be taken into account in the evaluation of postmenopausal and male osteoporosis. Although severe vitamin D deficiency leading to rickets or osteomalacia is rare in Brazil, there is accumulating evidence of the frequent occurrence of subclinical vitamin D deficiency, especially in elderly people. PMID- 17117290 TI - Asymptomatic primary hyperparathyroidism. AB - Primary hyperparathyroidism is a common disorder of mineral metabolism characterized by incompletely regulated, excessive secretion of parathyroid hormone from one or more of the parathyroid glands. In adults with the disease, a single, benign adenoma is seen approximately 80 percent of the time, with multiple gland involvement comprising most of the remaining patients. Very rarely, a parathyroid cancer is responsible but it is seen in less than 0.5 percent of patients with primary hyperparathyroidism. In this article, we will review important clinical and diagnostic features of asymptomatic primary hyperparathyroidism as well as considerations for surgical or medical management of the disease. PMID- 17117291 TI - From mild to severe primary hyperparathyroidism: The Brazilian experience. AB - Primary hyperparathyroidism often presents as an asymptomatic disorder. In our institution, routine serum calcium measurements have now been used as part of medical examination for 23 years. Out of 124 patients consecutively seen at our institution, 47% presented with no symptoms related to the disease, while 25% presented with severe skeletal involvement and osteitis fibrosa cystica, 25% with renal stone disease without overt bone involvement, and 2% with the typical neuropsychiatric syndrome. This same pattern is seen in the city of Sao Paulo. In severe disease pathological fractures are frequently seen, especially in long bones of the lower extremities, and also loss of lamina dura of the teeth and salt-and-pepper appearance of the skull. Bone mineral density is extremely low in these patients but usually show remarkable recovery following surgical cure. Serum PTH and bone markers are considerable higher in severely affected patients, who also have a high rate of vitamin D deficiency, and the parathyroid lesion is easier located compared with asymptomatic patients. From pathological specimens 87% had histological confirmation of a single adenoma, 6.4% multiple gland hyperplasia and 3.8% carcinoma. PMID- 17117292 TI - Hypoparathyroidism and pseudohypoparathyroidism. AB - The principal function of the parathyroid hormone (PTH) is maintenance of calcium plasmatic levels, withdrawing the calcium from bone tissue, reabsorbing it from the glomerular filtrate, and indirectly increasing its intestinal absorption by stimulating active vitamin D (calcitriol) production. Additionally, the PTH prompts an increase in urinary excretion of phosphorus and bicarbonate, seeking a larger quantity of free calcium available in circulation. Two mechanisms may alter its function, limiting its control on calcium: insufficient PTH production by the parathyroids (hypoparathyroidism), or a resistance against its action in target tissues (pseudohypoparathyroidism). In both cases, there are significantly reduced levels of plasmatic calcium associated with hyperphosphatemia. Clinical cases are characterized by nervous hyperexcitability, with paresthesia, cramps, tetany, hyperreflexia, convulsions, and tetanic crisis. Abnormalities such as cataracts and basal ganglia calcification are also typical of these diseases. Treatment consists of oral calcium supplementation associated with increased doses of vitamin D derivatives. PMID- 17117293 TI - Office practice of osteoporosis evaluation. AB - Osteoporosis is a metabolic disease characterized by low bone mass and microarchitectural deterioration of bone tissue, leading to enhanced bone fragility and a consequent increase in fracture risk. Bone fragility depends on bone density, turnover and microarchitectural features, such as relative trabecular volume, spacing, number and connectivity. Previous fragility fractures increase the fracture risk irrespective of bone density. Other risk factors must also be considered as many fractures occur in patients with osteopenia on densitometry. On the other hand, the diagnosis of osteoporosis and increased fracture risk should not be based on densitometric data alone when young populations such as men below 65 years, premenopausal women, adolescents and children are considered. PMID- 17117294 TI - Calcium intake and disease prevention. AB - While the fundamental metabolic function of calcium is to serve as a second messenger, coupling intracellular responses to extracellular signals, nutritional deficiency of calcium is manifested at a higher level of organization: 1) depletion of the calcium nutrient reserve; 2) inadequate complexation of digestive byproducts; and 3) collateral effects of hormones produced primarily to compensate for low calcium intake. The first mechanism contributes to the osteoporosis problem, the second to kidney stones and colon cancer, and the third to hypertension, preeclampsia, obesity, and insulin resistance, among others. Adequate calcium intakes (1000-1500 mg/d) in adults have been shown in controlled trials to lower the risk of osteoporotic fractures, kidney stones, obesity, and hypertension. The best source of calcium is dairy foods, largely because the disorders concerned depend upon multiple nutrients, not just calcium, and dairy provides a broad array of essential nutrients in addition to calcium, and at low cost. PMID- 17117295 TI - Redefining osteoporosis treatment: Who to treat and how long to treat. AB - Osteoporosis is a common disease that is associated with increased risk of fractures and serious clinical consequences. Bone mineral density (BMD) testing is used to diagnose osteoporosis, estimate the risk of fracture, and monitor changes in BMD over time. Combining clinical risk factors for fracture with BMD is a better predictor of fracture risk than BMD or clinical risk factors alone. Methodologies are being developed to use BMD and validated risk factors to estimate the 10-year probability of fracture, and then combine fracture probability with country-specific economic assumptions to determine cost effective intervention thresholds. The decision to treat is based on factors that also include availability of therapy, patient preferences, and co-morbidities. All patients benefit from nonpharmacological lifestyle treatments such a weight bearing exercise, adequate intake of calcium and vitamin D, fall prevention, avoidance of cigarette smoking and bone-toxic drugs, and moderation of alcohol intake. Patients at high risk for fracture should be considered for pharmacological therapy, which can reduce fracture risk by about 50%. PMID- 17117296 TI - Estrogen therapy for postmenopausal osteoporosis. AB - Osteoporosis is a worldwide problem that results in fractures that lead to disability and high costs to society. Estrogen therapy is frequently utilized for postmenopausal symptoms, but also has proven protective effects on the skeleton. The main action of estrogen at the cellular level is to inhibit the osteoclast by increasing levels of osteoprotegerin (OPG). OPG binds to the receptor activator of NFkB and prevents osteoclast differentiation, activity and survival. Numerous trials have demonstrated the positive effect estrogen has on the improvement of bone mineral density, and lower doses have also proven efficacious with fewer side effects. Both observational and randomized clinical trials have demonstrated the ability of estrogen treatment to prevent fractures. Topics that remain controversial include the appropriate length of estrogen treatment for postmenopausal women and the appropriate follow-up after treatment discontinuation. PMID- 17117297 TI - Selective estrogen receptor modulators (SERMS). AB - Hormone receptors and, specifically, estrogen receptors were described about four decades ago. For estrogens, there are two receptors, estrogen receptor alpha (ERalpha) and estrogen receptor beta (ERbeta). The two receptors are coded by different genes and their tissue expression varies across organs. ERalpha is predominantly expressed in reproductive tissues (uterus, breast, ovaries) liver and central nervous system, whereas ERbeta is expressed in other tissues such as bone, endothelium, lungs, urogenital tract, ovaries, central nervous system and prostate. More than seventy molecules that belong to the SERMS class have been described. There are 5 chemical groups: triphenylethylenes, benzotiophenes, tetrahydronaphtylenes, indoles and benzopyrans. All of these non-hormonal compounds are capable of activating the ER, reduce bone turnover rate and, as an antiresorptive, clearly improve bone density. Estrogens reduce bone turnover rate and, as an antiresorptive, clearly improve bone density. They are also beneficial for the relief of menopausal symptoms. An ongoing debate that extends over the decades, relates to to overall benefit/risk profile of estrogen or estrogen progestin therapy since these therapies can increase the risk of serious health disorders, such as breast cancer. SERMs have increased our understanding of hormone-receptor regulatory mechanisms. Their development has permitted a targeted efficacy profile avoiding some of the side effects of the hormone therapy. Their clinical utility relies today mostly on the effects on breast cancer and bone. PMID- 17117298 TI - Bisphosphonates. AB - Osteoporosis is the result of bone loss due to an imbalance in bone turnover such that bone resorption exceeds bone formation. Bisphosphonates are potent inhibitors of osteoclast activity that reduce bone turnover and re-establish the balance between bone resorption and formation. In clinical studies, several bisphosphonates prevent bone loss, preserve bone structure, improve bone strength and, in patients with osteoporosis, substantially reduce fracture risk. They are effective in multiple clinical settings including postmenopausal osteoporosis, low bone mass in men and drug-induced bone loss. Intermittent oral dosing and intravenous administration are more convenient than the original daily dosing regimen. These drugs are generally well tolerated and have an excellent safety profile in that serious side effects are uncommon. Potent bisphosphonates are generally the preferred treatment option for most patients with or at risk for osteoporosis. PMID- 17117299 TI - Anabolic skeletal therapy for osteoporosis. AB - Antiresorptive agents for osteoporosis are a cornerstone of therapy, but anabolic drugs have recently widened our therapeutic options. By directly stimulating bone formation, anabolic agents reduce fracture incidence by improving bone qualities besides increasing bone mass. In this article, we review the role of anabolic treatment for osteoporosis. The only anabolic agent currently approved in the United States for osteoporosis, teriparatide [recombinant human parathyroid hormone(1-34)], has clearly emerged as a major approach to selected patients with osteoporosis. Teriparatide increases bone density and bone turnover, improves microarchitecture, and changes bone size. The incidence of vertebral and nonvertebral fractures is reduced. Teriparatide is approved for both postmenopausal women and men with osteoporosis who are at high risk for fracture. Other potential anabolic therapies for osteoporosis, including other forms of parathyroid hormone, strontium ranelate, growth hormone, and insulin-like growth factor-1, are also reviewed in this article. PMID- 17117300 TI - Update on osteoporosis therapy. AB - Osteoporosis is a skeletal disorder characterized by compromised bone strength that predisposes a person to increased fracture risk. Fractures are often associated with increased morbidity, higher mortality, loss of function and even psychological consequences. Pharmacotherapeutic interventions (e.g., bisphosphonates, selective estrogen receptor modulators, calcitonin, and teriparatide) in women with postmenopausal osteoporosis provide substantial reduction in fracture risk over and above risk reduction with calcium and vitamin D supplementation alone. The importance of nutritional support along with an appropriate exercise regimen, avoiding smoking and excessive alcohol use is to be emphasized along with the pharmacologic approach to osteoporosis. Despite the effectiveness of therapy with pharmacologic agents, most patients who start therapy do not remain on treatment for more than 1 year. PMID- 17117301 TI - Osteoporosis in men. AB - Osteoporosis is defined as "a systemic skeletal disease characterized by low bone mass and microarchitectural deterioration of bone tissue with a consequent increase in bone fragility and susceptibility to fracture". Approximately 40-50% of women sustain osteoporotic fractures in their lifetime; as such, it is appropriate that studies initially focused upon females. Despite an increased recognition of osteoporotic fractures in men, there continues to be neglect of this disease in males. This ongoing neglect is inappropriate as 25-33% of men in some populations will sustain osteoporotic fractures in their lifetime. Testosterone plays an important role in male skeletal health. However, recent data suggest that estrogen may in fact be the dominant hormone regulating skeletal status in both men and women. BMD measurement may be utilized for osteoporosis diagnosis and to assist with fracture risk prediction in men prior to their sustaining a fracture. Recognizing this need, the International Society for Clinical Densitometry (ISCD) recommended and recently reaffirmed use of a BMD T-score of -2.5 or below be utilized to diagnose osteoporosis in men. Androgen therapy of hypogonadal men may be considered with the caveat that data do not exist to document that this treatment reduces fracture risk. At this time, the data is inadequate to support use of androgen treatment in eugonadal men with osteoporosis. Parathyroid hormone treatment does increase BMD; existing studies have not been of adequate size or duration to document fracture reduction efficacy. Bisphosphonate therapy increases BMD, reduces vertebral fracture risk and is considered the standard of care for osteoporotic men at this point in time. PMID- 17117302 TI - Low bone mass in children and adolescents. AB - Osteoporosis is a disease characterized by low bone mass and micro architectural alterations of bone tissue leading to enhanced bone fragility and increased fracture risk. Although research in osteoporosis has focused mainly on the role of bone loss in the elderly population, it is becoming increasingly clear that the amount of bone that is gained during growth is also an important determinant of future resistance to fractures. Thus, considerable interest is being placed on defining preventive strategies that optimize the gain of bone mass during childhood and adolescence. Knowledge of the determinants accounting for the physiologic and genetic variations in bone accumulation in children will provide the best means toward the early diagnosis and treatment of osteoporosis. This article reviews the techniques available for bone mass measurements in children and the major determinants and diseases influencing bone accretion during childhood and adolescence. PMID- 17117303 TI - Transplantation osteoporosis. AB - In the past two decades, there has been a rapid increase in the number of organ transplanted worldwide, including Brazil, along with an improvement in survival and quality of life of the transplant recipients. Osteoporosis and a high incidence of fragility fractures have emerged as a complication of organ transplantation. Many factors contribute to the pathogenesis of osteoporosis following organ transplantation. In addition, most patients have some form of bone disease prior to transplantation, which is usually related to adverse effects of end-stage organ failure on the skeleton. This chapter reviews the mechanisms of bone loss that occur both in the early and late post-transplant periods, as well as the features specific to bone loss after kidney, lung, liver, cardiac and bone marrow transplantation. Prevention and treatment for osteoporosis should be instituted prior and in the early and late phase after transplantation, and will also be addressed in this article. PMID- 17117304 TI - Glucocorticoid-induced osteoporosis. AB - Glucocorticoid-induced osteoporosis is the most frequent cause of secondary osteoporosis. Glucocorticoids cause a rapid bone loss in the first few months of use, but the most important effect of the drug is suppression of bone formation. The administration of oral glucocorticoid is associated with an increased risk of fractures at the spine and hip. The risk is related to the dose, but even small doses can increase the risk. Patients on glucocorticoid therapy lose more trabecular than cortical bone and the fractures are more frequent at the spine than at the hip. Calcium, vitamin D and activated forms of vitamin D can prevent bone loss and antiresorptive agents are effective for prevention and treatment of bone loss and to decrease fracture risk. Despite the known effects of glucocorticoids on bone, only a few patients are advised to take preventive measures and treat glucocorticoid-induced osteoporosis. PMID- 17117305 TI - Hypophosphatemic rickets and osteomalacia. AB - The hypophosphatemic conditions that interfere in bone mineralization comprise many hereditary or acquired diseases, all of them sharing the same pathophysiologic mechanism: reduction in the phosphate reabsorption by the renal tubuli. This process leads to chronic hyperphosphaturia and hypophosphatemia, associated with inappropriately normal or low levels of calcitriol, causing osteomalacia or rickets in children and osteomalacia in adults. X-linked hypophosphatemic rickets, autosomal-dominant hypophosphatemic rickets, and tumor induced osteomalacia are the main syndromes involved in the hypophosphatemic rickets. Although these conditions exhibit different etiologies, there is a common link among them: increased activity of a phosphaturic factor, being the fibroblast growth factor 23 (FGF-23) the most studied one and to which is attributed a central role in the pathophysiology of the hyperphosphaturic disturbances. Activating mutations of FGF-23 and inactivating mutations in the PHEX gene (a gene on the X chromosome that codes for a Zn-metaloendopeptidase proteolytic enzyme which regulates the phosphate) involved in the regulation of FGF-23 have been identified and have been implicated in the pathogenesis of these disturbances. Genetic studies tend to show that the phosphorus homeostasis depends on a complex osteo-renal metabolic axis, whose mechanisms of interaction have been poorly understood so far. This paper reviews the current knowledge status concerning the pathophysiology of phosphate metabolism regulation and the pathophysiologic basis of hypophosphatemic rickets. It also analyzes the clinical picture and the therapeutic aspects of these conditions as well. PMID- 17117306 TI - Paget's disease of bone. AB - Paget's disease of bone is a focal disorder of bone remodeling accompanied initially by an increase in bone resorption, followed by a disorganized and excessive formation of bone, leading to pain, fractures and deformities. It exhibits a marked geographical variation in its prevalence. In Brazil it predominantly affects persons of European descent. The majority of the reported cases of the disease in Brazil are from Recife, owing to its peculiar mixed European colonization over approximately four centuries. The etiology is complex and involves both genetic and environmental factors. The disease is often asymptomatic and diagnosis is usually based on biochemical markers of bone turnover, radionuclide bone scan and radiological examination. Bisphosphonates, in particular zoledronic acid, are regarded as the treatment of choice for Paget's disease of bone. PMID- 17117307 TI - Renal stone disease: Causes, evaluation and medical treatment. AB - The purpose of the present review is to provide an update about the most common risk factors or medical conditions associated with renal stone formation, the current methods available for metabolic investigation, dietary recommendations and medical treatment. Laboratory investigation of hypercalciuria, hyperuricosuria, hyperoxaluria, cystinuria, hypocitraturia, renal tubular acidosis, urinary tract infection and reduction of urinary volume is based on the results of 24-hr urine collection and a spot urine for urinary sediment, culture and pH. Blood analysis for creatinine, calcium and uric acid must be obtained. Bone mineral density has to be determined mainly among hypercalciurics and primary hyperparathyroidism has to be ruled out. Current knowledge does not support calcium restriction recommendation because it can lead to secondary hyperoxaluria and bone demineralization. Reduction of animal protein and salt intake, higher fluid intake and potassium consumption should be implemented. Medical treatments involve the use of thiazides, allopurinol, potassium citrate or other drugs according to the metabolic disturbances. The correction of those metabolic abnormalities is the basic tool for prevention or reduction of recurrent stone formation. PMID- 17117308 TI - Culture, moral experience and medicine. AB - No one can doubt any longer that culture is crucial to medicine. The evidence for health disparities across ethnic and racial groups as well as for cultural influences on health care practices is too impressive to overlook. Yet the concept of culture and how it is employed in medicine today is quite different from the way culture is now regarded in anthropology, the discipline that originated and popularized the concept. Rather than understand culture as a "timeless" ethnic stereotype applied to patients-which is a common but dangerous practice-physicians need to understand how culture influences doctors as much as patients. And physicians need to understand that culture is not only about differences in dress, etiquette and diet, but also and most profoundly, about what really matters to people. That is, culture is about the changing moral experiences of patients, families, and practitioners, and how those moral experiences powerfully affect the doctor-patient relationship. This article suggests that there is a moral crisis in today's medicine that reflects global cultural transitions. This crisis must be addressed if practitioners are to provide care at the highest moral and human level. PMID- 17117309 TI - Left ventricular remodeling after myocardial infarction: past, present, and future. AB - Medical advances in the care of cardiac patients have resulted in more patients surviving an acute myocardial infarction (MI) than ever before. Each year hundreds of thousands of these survivors undergo remodeling of their left ventricle and often progress to clinical congestive heart failure. The extent of remodeling has been linked to the size of the infarct, whether or not the myocardium has been revascularized, and the control of loading conditions. The extent of infarction can be measured several ways, including the amount of enzyme released as well as infarct imaging with nuclear perfusion or magnetic resonance imaging. Methods to prevent adverse remodeling of the ventricle have included pharmacotherapy with beta-blockers, nitrates, and modulators of the renin angiotensin-aldosterone system. Surgical intervention has proven useful for select patients with aneurysmal areas of remodeling. Researchers are now investigating several approaches to preventing and reversing cardiac remodeling. These include the use of stem cells to regenerate myocardium and post-infarct pacing to prevent remodeling. Improved therapies are needed to help reduce the number of patients progressing from myocardial infarction to end-stage heart failure. PMID- 17117310 TI - Prevention and management of occupational exposures to human immunodeficiency virus (HIV). AB - Occupational exposure to blood and other potentially infectious body fluids places health care workers at risk for acquisition of bloodborne pathogens, including the human immunodeficiency virus (HIV). Utilizing appropriate techniques, personal protective equipment, and safer "sharp" technology can minimize the risk of these exposures. When exposure does occur, immediate evaluation and initiation of post-exposure prophylaxis, when indicated. can substantially reduce the risk of transmission of HIV. In this article, the basic concepts of exposure prevention and management are reviewed. PMID- 17117311 TI - The changing library: what clinicians need to know. AB - Over the last two decades, changes in technology have allowed academic medical center libraries to bring the world of biomedical information to the physician's computer desktop. Because digital libraries have grown so rapidly and in so many ways, some clinicians may be uncertain about the services and resources that are available to them. This article explains how clinical faculty can best utilize their library to support their research and patient care. It addresses some of the most common myths about the "new" medical library, and it highlights innovations in library resources and services that can help physicians to better access, use and manage medical information. PMID- 17117312 TI - Percutaneous endoscopic lumbar discectomy for adolescent lumbar disc herniation: surgical outcomes in 46 consecutive patients. AB - BACKGROUND: The surgical outcome of percutaneous endoscopic lumbar discectomy (PELD) for adolescent lumbar disc herniation has rarely been reported on. We performed this study to evaluate the surgical outcome of PELD for adolescent lumbar disc herniation. METHODS: We analyzed the surgical outcomes in 46 consecutive adolescent patients between 13 and 18 years of age (mean age, 16.5 years) who underwent PELD for single level lumbar disc herniation from June 2000 to May 2002. Using the clinical charts and mailed questionnaires, we evaluated the patients preoperatively by the postoperative Visual Analogue Scale (VAS) for back and leg pain, and by the postoperative Macnab criteria. RESULTS: PELD was performed at L3-4 on one patient, at L4-5 on 40 patients and at L5-S1 on 5 patients. One patient complained of transient dysesthesia after the operation. Another patient underwent subsequent open discectomy because only incomplete decompression was achieved with PELD. At a mean follow-up duration of 37.2 months (range: 25-48 months), the mean VAS scores of both the back and leg pain decreased significantly. In terms of the Macnab criteria, 91.3% of the patients showed excellent or good outcomes. Recurrent disc herniation developed in one patient 14 months after surgery. CONCLUSIONS: Adolescents who underwent PELD for single level soft lumbar disc herniation showed favorable results that were comparable to the results of open discectomy. PMID- 17117313 TI - Benign splenic epithelial cyst accompanied by elevated Ca 19-9 level: a case report. AB - We report the case of a 30-year-old woman with a benign epidermoid splenic cyst and a high CA 19-9 serum level (268 U/mL). The patient underwent resection of the cyst and splenectomy. After removal of the cyst, the serum CA 19-9 level decreased to a normal level within 6 weeks. True non-parasitic splenic cysts are rare. Approximately 30 cases of benign true splenic cysts with a high CA 19-9 serum level have been published. PMID- 17117314 TI - Surgical management of functional tricuspid regurgitation with a new remodeling annuloplasty ring. AB - BACKGROUND: Moderate-to-severe functional tricuspid regurgitation (TR) should be corrected in patients undergoing surgery for left-sided valvular diseases, to improve long-term outcomes. Several techniques of surgical repair (suture annuloplasty or prosthetic annuloplasty) to correct this condition have been described. Multiple clinical studies have shown the superiority of prosthetic remodeling annuloplasty over the other surgical approaches. Despite this, suture based annuloplasty remains the most commonly used technique for tricuspid valve repair. A new 3-dimensional remodeling prosthesis has been developed to address the issue of residual TR. We report our early experience with this new 3 dimensional prosthetic remodeling ring, the Edwards MC 3 system. MATERIAL: From August 2002 to March 2004, 51 patients (24 male, 27 female, mean age 64 +/- 15, ejection fraction 49 +/- 15, median NYHA III [II-IV]) underwent tricuspid valve repair for functional TR due to annular dilatation, with the Edwards MC 3 system. Etiology of left-sided valvular disease was: rheumatic (n = 19), degenerative (n = 16), ischemic cardiomyopathy (n = 1), and endocarditis (n = 5). Twenty (50%) patients underwent redo operations. Concomitant procedures included: mitral valve surgery (repair n = 34, replacement n = 14), aortic valve replacement (n = 5), coronary artery bypass graft (n = 8) and left arterial maze (n = 16). Median EuroSCORE was 12% (1-74%) in this patient population. RESULTS: Operative and late mortality were 3.8% (n = 2) and 13.7% (n = 7), respectively. Echocardiography at discharge showed a mean TR decrease from 3.1 +/- 0.9 to 0.3 +/- 0.4 (p < 0.001) and mean mitral regurgitation (MR) decrease from 3.2 +/- 1 to 0.1 +/- 0.1 (p < 0.001), while ejection fraction increased to 53% (p = 0.047), and at 6-month follow-up, mean TR and MR remained unchanged. CONCLUSION: Concomitant tricuspid valve repair for functional TR with left-sided valve surgery carries a low operative mortality. The Edwards MC 3 annuloplasty system is relatively simple to implant and corrects TR effectively (without significant residual TR), while providing excellent short-term clinical results. The 3-dimensional saddle shape of this ring may further optimize the fixation of the annulus in systolic position, and improve long-term results. Larger clinical series with longer-term follow-up are necessary to confirm these early promising results. PMID- 17117315 TI - Rapid ventricular pacing due to electrocautery: a case report and review. AB - It is well known that the use of electrocautery during surgery can interfere with the functioning of pacemakers. The most common problem is that the electrocautery is interpreted by the pacemaker as cardiac electrical activity, which inhibits the pacemaker from firing, leading to bradycardia. The use of electrocautery in a site remote from the pacemaker generator normally does not cause interference. It is also very unusual to develop a tachycardic response to the electrocautery. We report a case of electrocautery causing rapid pacing during transurethral resection of bladder polyps. The possible causes of inappropriate rapid pacing are reviewed. PMID- 17117316 TI - Late recurrent pulmonary typical carcinoid tumor: case report and review of the literature. AB - Carcinoid tumors are uncommon pulmonary neoplasms. They are classified histologically as either atypical or typical. Atypical carcinoids are aggressive malignancies that require radical surgical resection and have a guarded prognosis with a propensity to metastasize and recur. Typical carcinoids are low-grade malignancies with relatively less metastatic or recurring potential and are usually treated with simple excision. Recurrence of a typical pulmonary carcinoid tumor more than a decade after initial resection is very rare. A patient with recurrence of a typical carcinoid tumor 11 years after resection of the primary lesion with one involved lymph node is reported here. Late recurrences are rare in both atypical and typical varieties, but are much more common in atypical carcinoids. The patient reported here represents the fifth case of recurrence of a typical carcinoid tumor more than ten years after resection. This suggests that, after resection of a typical carcinoid neoplasm, patients should be monitored carefully, especially if lymph node metastases are present at the time of surgery. PMID- 17117317 TI - Intestinal tuberculosis and secondary liver abscess. AB - The incidence of intestinal tuberculosis (ITB) has been increasing in the West, due to the AIDS epidemic, transglobal immigration, IV drug abuse, an aging population, and an increase in the number of immunocompromised patients. Obstruction and perforation of the intestine are the most common and serious complications of ITB. Another complication, tuberculous liver abscess (TLA), is rare and usually associated with foci of infection in the lung or gastrointestinal tract. We report a case of a 17-year-old boy with Down syndrome who presented with multiple TLAs secondary to obstructive and multiple perforated ileal tuberculosis. PMID- 17117318 TI - Black hairy tongue associated with olanzapine treatment: a case report. AB - Olanzapine is an atypical antipsychotic drug approved for acute and long-term treatment of bipolar disorder. Although relatively safe as compared to other classical antipsychotic medications, there are a number of uncommon adverse effects of olanzapine such as oral cavity lesions. In addition to the relatively common side effect of dry mouth, several articles have reported an association between olanzapine treatment and the development of oral lesions such as apthous stomatitis, pharyngitis, glossitis and oral ulceration. Although there are several cases in which the tongue was affected in conjunction with stomatitis or pharyngitis, we could not find a case report indicating a direct relationship between olanzapine use and a tongue lesion. We present here the case of a patient with bipolar disorder, who developed recurrent black hairy tongue on two different occasions following the addition of olanzapine to lithium treatment. In the present case, xerostomia (dry mouth), which is an adverse reaction of both olanzapine and lithium, may have played a role in the development of black hairy tongue. All agents with a possible side effect of xerostomia may predispose patients to black hairy tongue, especially when they are administered in combination. To preclude the development of this complication with such drugs, extra time and effort should be given to improving oral hygiene. PMID- 17117319 TI - Recurrent episodes of dermatomyositis-associated pneumonitis masquerading as hypersensitivity pneumonitis. AB - This case report describes a unique presentation of dermatomyositis-associated pneumonitis. A 44-year-old man presented with repeated episodes of fever, dyspnea, and hemoptysis accompanied with pulmonary infiltrates, on chest CT scan. Hypersensitivity pneumonitis was suspected. Further work-up showed clinical and serologic evidence of dermatomyositis-associated pneumonitis. The patient was treated with oral prednisone and azathioprine. The subject of dermatomyositis associated pneumonitis is discussed. PMID- 17117320 TI - Metastatic signet ring adenocarcinoma: an unusual cause of cardiac constriction. AB - Pericardial constriction secondary to metastatic adenocarcinoma is exceedingly rare. We present the first recorded case of pericardial constriction secondary to metastatic signet-ring mucinous adenocarcinoma diagnosed by echocardiography. The cornerstones of echocardiographic diagnosis of constriction are the following: interventricular septal bounce phasic with respiration, M-mode recordings of the inferior vena cava, and the characteristic Doppler velocity patterns recorded from the mitral valve, hepatic veins, and mitral annulus. PMID- 17117321 TI - Bronchopulmonary sequestration--a 12-year experience. AB - Bronchopulmonary sequestration (BPS) is a rare malformation of the lower respiratory tract. It consists of a non-functioning mass of lung tissue that lacks normal communication with the tracheobronchial tree and that receives arterial blood supply from the systemic circulation. It is classified as intralobar (ILS) or extralobar (ELS) according to its location within or outside the normal lung. Most sequestrations are intralobar (75%) and occur predominantly in the left lower lobe. PMID- 17117322 TI - CT-guided percutaneous transthoracic biopsy in the evaluation of undetermined pulmonary lesions. AB - CT-guided Percutaneous Transthoracic Biopsies (PTB) performed in the Radiology Department of Garcia de Orta Hospital between 2002 and 2004 to evaluate undetermined pulmonary lesions were retrospectively analysed. 89 fine needle aspiration biopsies (FNAB) and 13 core needle biopsies (CNB) were performed on 92 patients (67 men, mean age: 64.4 years). 82 lesions (89%) were nodular lesions (mean diameter: 3.8+/-1.7 cm, 65 peripheral). We did not observe complications among patients who underwent CNB; minor complications and pneumothorax requiring drainage occurred in 11 FNAB. 72 FNAB were considered adequate for cytology diagnosis; 72% of them positive for malignancy. All CNB were adequate and conclusive. From the 7 CNB performed on patients with previous FNAB, 3 allowed a better histological characterization and in 3 cases of inadequate FNAB, CNB was conclusive. All malignant lesions were nodules: 20 adenocarcinoma, 13 non-small cell lung cancer (SCLC), 10 epidermoid tumours, 5 small-cell lung cancer, 2 carcinoids, 1 bronchiolo alveolar carcinoma, 1 malignant mesothelioma and 8 metastasis. Unspecific/inflammatory lesions (n=5) were the most frequent benign lesions. Malignant lesions were more prevalent in older patients (p=0.007) and were larger (p=0.006). Spiculated and lobulated contour (p=0.05) were more prevalent in malignant lesions while regular contour was more frequent among benign lesions (p=0.0001). Gender, smoking, location, pleural tag, homogenous attenuation, cavitation, calcification, necrosis and air bronchogram did not differ significantly between benign and malignant nodules. This study shows that CT-guided PTB is a safe and effective procedure in the evaluation of undetermined pulmonary lesions. PMID- 17117323 TI - Smoking trends in Portuguese school-aged children and approaches for a control- an analysis based on the Health Behaviour in School-Aged Children (HBSC) data. AB - While smoking has negative health consequences for children and adolescents, the major risk of smoking onset by these age groups is tobacco dependence, in most cases for the rest of their lives, which can later lead to the suffering of diseases related to smoking. This fact shows the importance of smoking prevention in teens to avoid the negative health, economic and environmental effects related to smoking. Although Portugal does not have a National Smoking Prevention Programme, some schools have developed prevention campaigns to control the spread of the tobacco epidemic. To determine the efficacy of smoking prevention campaigns developed in Portuguese schools we compared the data of smoking prevalence provided by Health Behaviour in School-Aged Children (HBSC) (11-15 years old) from 1997/98 and 2002. The results show the rise of smoking prevalence in students on both sexes, especially among girls. Data supports the conclusion that Portuguese schools are not effective in smoking prevention and in Portugal it is necessary to continue smoking prevention campaigns aimed at the younger children. PMID- 17117324 TI - Respiratory squamous cell carcinomas in vibroacoustic disease. AB - BACKGROUND: In 1987, the autopsy of a vibroacoustic disease (VAD) patient disclosed two tumours: a renal cell carcinoma and a malignant glioma in the brain. Since 1987, malignancy in VAD patients has been under close surveillance. To date, in a universe of 945 individuals, there are 46 cases of malignancies, of which 11 are multiple. Of the 11 cases of respiratory tract tumours, all were squamous cell carcinomas (SqCC). This report focuses on the morphological features of these tumours. METHODS: Tumour fragments were collected (endoscopic biopsy or surgery) from 11 male VAD patients (ave. age: 58+/-9 years, 3 non smokers): 2 in glottis and 9 in the lung. In the 3 non-smokers, 2 had lung tumours and 1 had a glottis tumour. All were employed as or retired aircraft technicians, military or commercial pilots. Fragments were fixed either for light and electron microscopy. Immunohistochemistry studies used chromagranine and synaptophysine staining. RESULTS: All lung tumours were located in the upper right lobe bronchi and were histologically poorly differentiated SqCC (Figs. 1, 2). The search with neuroendocrine markers was negative. The average age of tumour onset in helicopter pilots was below 50 years old while for the other professional groups it was above 50. Nine patients are deceased. The 2 surviving patients are heavy smokers (> 2 packs/day). Smoking habits had no influence on tumour outcome and progression. DISCUSSION: Epidemiological studies indicate that squamous cell carcinomas account for approximately 40% of all lung tumours in men. It seems to be highly relevant that all VAD patient respiratory tract tumours are squamous cell carcinomas. It is not surprising that helicopter pilots are the ones who are affected the earliest because previous studies have shown that helicopter pilots exhibited the highest values for the frequency of sister chromatid exchanges. Generally, epidemiological tumor studies do not take histological tumor type into account, but given the results herein, it would seem of the utmost importance to begin specifiying the exact histological type of tumor in all statistical studies. PMID- 17117325 TI - [Lung cancer in women: a different entity?]. AB - Lung cancer is becoming a real epidemic in developed countries and one of the main causes of cancer death in women. Although it is controversial to state, it is probable that women are more susceptible to lung cancer than men. Molecular and genetic epidemiology studies are underway to prove this statement. Several biological factors, such as family history, histopathology, response to treatment and the prognosis for lung cancer in women have a bearing on the case. While these are not completely clear or consensual, there is a need for wide-ranging prospective studies which compare the differences between males and females. Until now, there has only been one area in which gender could impact on the therapeutic management of lung cancer: the role of gefitinib and erlotinib in inhibiting the epidermal growth factor receptors, since these products are clearly of more benefit to female non-smokers. Given that women have a better lung cancer prognosis, it is recommended that future research protocols include stratification on gender. Prevention of lung cancer in both women and men is a priority public health concern. A mandatory aim of this is the fight against smoking, the largest aetiological factor of lung cancer. PMID- 17117326 TI - [Allergic inflammatory diseases of the upper airways and their impact on asthma- following on from a case report]. AB - The authors present a case report of a patient referred to physician for "difficult-to-treat asthma". Clinical evaluation concluded that severe chronic inflammatory upper airway disease was an asthma worsening factor. Patient demonstrated improvement in asthma control, after surgery. In the context of the clinical study, the authors present a review of the most common allergic inflammatory diseases of the upper airways (allergic rhinitis, chronic sinusitis and nasal polyposis) and reflect on their impact on asthma. PMID- 17117327 TI - [Interstitial idiopathic pneumonias--a literature review]. AB - Interstitial idiopathic pneumonias are a group of diseases whose rarity and variety of clinical, radiological and pathological descriptions creates difficulties in study and management. An example of this is the classification method for this group, with only 2002 seeing some consensus. The aim of this article is to review the main literature to contribute to an understanding of this subject. PMID- 17117328 TI - [Guidelines for the diagnosis and treatment of latent tuberculosis infection and active tuberculosis in patients with inflammatory joint diseases proposed for treatment with tumour necrosis factor alpha antagonist drugs]. AB - The Portuguese Society of Rheumatology (SPR) and the Portuguese Society of Pulmonology (SPP) have developed guidelines for the diagnosis and treatment of latent tuberculosis infection (LTBI) and active tuberculosis (AT) in patients with inflammatory joint diseases (IJD), namely rheumatoid arthritis, psoriatic arthritis and ankylosing spondylitis, treated with tumour necrosis factor alpha (TNF-a) antagonists. Due to the high risk of tuberculosis (TB) in patients with IJD, LTBI and AT screening should be performed as soon as possible, ideally at the moment of IJD diagnosis. Even if TB screening was performed at the beginning of the disease, the evaluation should be repeated before starting anti-TNF-a therapy. When TB (LTBI or AT) treatment is indicated, it should be performed before the beginning of anti-TNF-a therapy. If the IJD activity requires urgent anti-TNF-a therapy, these drugs can be started after two months of antituberculosis therapy in AT cases, or after one month in LTBI cases. Chest X ray is mandatory for all patients. If abnormal, e.g. Gohn complex, the patient should be treated as LTBI; residual lesions require the exclusion of AT and patients with history of untreated or incomplete TB treatment should be treated as LTBI. In cases of suspected active lesions, AT diagnosis should be confirmed and adequate therapy initiated. Tuberculin skin test (TST), with two units of RT23, should be performed in all patients. If induration is less than 5 mm, the test should be repeated after 1 to 2 weeks, on the opposite forearm, and should be considered negative if the result is again inferior to 5 mm. Positive TST implicates LTBI treatment. If TST is performed in immunosuppressed IJD patients, LTBI treatment should be offered to the patient before starting anti-TNF-a therapy, even in the presence of a negative test. PMID- 17117329 TI - Acute fibrinous and organizing pneumonia. AB - The term Acute Fibrinous and Organizing Pneumonia (AFOP) has been proposed by Beasley et al for cases that do not fit into the histopathologic criteria of the recognized entities described as acute or subacute clinical presentations. The presence of intra-alveolar fibrin in the form of fibrin "balls" and organizing pneumonia with patchy distribution are the main histological features of this entity. We describe the case of a male patient with the diagnostic of AFOP made by surgical lung biopsy. He had a subacute presentation of symptoms consisting of productive cough, chest pain and fever. Bilateral infiltrates with patchy and diffuse distribution were the predominant features in his chest HRCT scan. The patient had a good clinical course after a treatment with prednisone and cyclophosphamide. Our hope in reporting this case study is to add some more data to the discussion of this new entity. PMID- 17117330 TI - [Pleomorphic carcinoma of the lung in a patient with Mounier-Kuhn syndrome]. AB - Pleomorphic carcinoma of the lung (PLC) is a rare malignant tumour presenting with a poor clinical outcome. It is included in the group of non-small cell lung carcinomas that contain sarcoma or sarcoma-like components (spindle cells/giant cells). Because of its rarity and diagnostic difficulties, the authors report a case of PLC in a 44 year-old patient, smoker, with the initial clinical presentation of a dorsal tumoural mass. As the time of diagnosis the tumour was in stage IV (TNM), the patient was treated only with supportive therapy and died five months later. During clinical evaluation, a Mounier-Kuhn Syndrome was confirmed. Clinical-imagiological aspects of this situation are reported as well as cytological and immunocytochemical features. PMID- 17117331 TI - [Depression in the elderly living in a rural area and other related factors]. AB - INTRODUCTION: We want to determine the prevalence of depression in a rural population (65 years and over) free from cognitive impairment and to evaluate related factors of late life depression. PATIENTS AND METHOD: A total of 265 persons (mean age [SD]: 76.2 [6.7] years; 60.4 % females) residing in the towns of Proaza, Quiros and Santo Adriano (Asturias) were interviewed. The evaluation included sociodemographical and clinical aspects as well as the Mini-Mental State Examination Spanish version (MMSE), the Geriatric Depression Scale (GDS), the Oviedo Sleep Questionnaire (OSQ), the CAGE Questionnaire, the Goldberg General Health Questionnaire (GHQ-28), and the World Health Organization Quality of Life Instrument (WHOQOL-BREF). RESULTS: A total of 23 subjects out of the total sample were excluded from the study due to scoring less than 18 points on the MMSE. The final sample included 242 subjects (mean age [SD]: 75.59 (6.2) years; 60.3 % females). Prevalence of probable depression was 23.1 % [30.1 % females vs 12.5 % males; p = 0.002]. Depression was statistically associated with a higher number of physical diseases (p = 0.012), higher psychiatric comorbidity (p = 0.031), less cognitive impairment measured by MMSE (p = 0.019), higher prevalence of sleep disorders (p < or = 0.050), higher score in the GHQ-28 (p = 0.000), and worse quality of life evaluated by the WHOQOL-BREF (p = 0.000). CONCLUSIONS: Depression is moderately prevalent in this population. Depression is more frequent in females, those living alone, without studies, working in agricultural sector, with physical or psychiatric comorbidity, with higher cognitive impairment, and with a worse health status and quality of life perception. PMID- 17117332 TI - [Clinical differences between morbid obese patients with and without binge eating]. AB - INTRODUCTION: Our objective was to evaluate clinical differences between morbid obese patients with and without binge eating. METHODS: We evaluated 210 morbid obese patients who were referred consecutively to a psychiatric evaluation in a general hospital for different reasons. We used a clinical interview, evaluated psychiatric comorbidity and applied a series of psychopathology and eating behavior scales: Beck Depression Inventory (BDI), Beck Anxiety Inventory (BAI), Maudsley Obsessive-Compulsive Interview (MOCI), Barrat Impulsiveness Scale (BIS), Eating Disorder Inventory (EDI), Eating Attitudes Test (EAT), Bulimic Investigatory Test Edinburgh (BITE) and Body Shape Questionnaire (BSQ). The sample was divided in two subgroups: patients with binge eating according to ICD 10 and DSM-IV criteria, and patients without binge eating. The first subgroup was composed of 14 patients with bulimia nervosa, 32 with binge eating disorder, and 15 patients with binge eating who did not fulfill criteria for bulimia or binge eating disorder (subgroup was composed by 61 patients vs 145 patients without binge eating). RESULTS: Binge eating patients showed more psychopathology, more prevalence of dysthymia, higher score in impulsiveness, and scales that evaluated eating disorder. Binge eating patients showed more familial background of eating disorders; more body dissatisfaction in spite of a lower Body Mass Index (BMI). This group showed more functional disability induced by obesity and more weight and shape overconcern. CONCLUSIONS. The findings suggest that the morbid obese with binge eating constitute a distinct subgroup (independently of eating disorder diagnosis) among the obese population, with more psychopathology severity, especially affective disorder, more impulsivity, and more severity in core items of eating disorder scales and body dissatisfaction. PMID- 17117333 TI - [Attitudes of Spanish doctors towards immigrant patients: an opinion survey]. AB - INTRODUCTION: We assess the attitudes of Spanish doctors towards immigrant patients. MATERIAL AND METHOD: DESIGN: cross-sectional descriptive study carried out in January-June 2003. SAMPLE AND SETTING: Family doctors and third-year residents of 15 health care centers in Zaragoza, Spain. INSTRUMENT: specifically designed interview answered in anonymously and on a voluntary basis. RESULTS: A total of 62.3 % of the sample responded. Nearly 75 % of the doctors had received training on immigration, mainly from a theoretical point of view. Almost 25 % of immigrant consultations are not related to health matters. Doctors think that immigrant patients somatize more than Spanish ones but they do not suffer from more psychiatric illness. They also think immigrants show less treatment compliance and are more demanding. They disagree about possible racial biases in diagnosis or treatment of these patients. Spanish doctors consider that the different ethnic groups present similar management difficulties. The group with the most negative stereotype was the patients from Muslim North African countries. DISCUSSION: These findings are discussed from a psychological and anthropological perspective. PMID- 17117334 TI - [Controversies about duration of untreated psychosis as independent prognostic variable of the evolutive course of schizophrenic psychoses]. AB - This study reviews recent literature on duration of untreated psychosis (DUP) and its most relevant characteristics and controversial issues, such as: a) why DUP has been pointed out as a main variable in first-episode psychosis research, and b) the role of DUP in designing intervention programs for the design and different action strategies in early intervention programs in psychoses. The authors also present data from a 2 year follow-up study of 231 patients with a diagnosis of schizophrenia and/or schizophreniform disorder (according to DSM-IV criteria). Results are included, analyzing DUP as prognostic factor for clinical outcome. Our conclusions suggest that DUP is a risk marker but not an independent prognostic factor determining follow-up in schizophrenic psychoses. Therefore, DUP's role in early intervention programs should be redefined. PMID- 17117335 TI - [Treatment adherence in schizophrenia. A comparison between patient's, relative's and psychiatrist's opinions]. AB - INTRODUCTION: Patients with psychiatric illness typically have great difficulty following a medication regimen, but they also have the greatest potential for benefiting from adherence. Due to the lack of insight in schizophrenia, adherence to treatment is especially important. We try to analyze and compare the opinion on adherence and compliance of psychiatrists, patients with schizophrenia and relatives. METHOD: A direct, anonymous survey specifically designed for the project was administered to psychiatrists, patients and relatives from all over Spain through different associations of patients and family legally constituted in Spain. Analysis was done separately for variables corresponding to the three groups. RESULTS: The psychiatrists (n = 844) considered that 56.8 % of their evaluated patients (n = 7.439) were noncompliers in the past month, as opposed to 43.2% of these patients who were considered good compliers (3,215 patients). Ninety-five percent of the patients (n = 938) stated that they took their medication regularly, while 5 % answered no to this question. Eighty-two percent of relatives (n = 796) think that patients regularly take their medication, but 47% state that they sometimes forget to take it. CONCLUSIONS: Treatment adherence should be evaluated in clinical trials and in research on treatment of diseases, particularly in chronic mental diseases such as schizophrenia. It seems clear that only programs aimed at detection and resolution of the problems involved in treatment adherence will be able to improve the mid- and long-term prognosis of patients with schizophrenic disorders. PMID- 17117336 TI - [Applicability of the ICF in measuring functioning and disability in unipolar depression in Primary Care settings]. AB - INTRODUCTION: We use the biopsychosocial model of the International Classification of Functioning, Disability, and Health (ICF): a) to analyze functioning and disability patterns in unipolar depression cases attended in primary care settings; b) to study predictive and mediator variables related to disability in depression, and c) to determine the impact of traditional interventions in depression cases using functional remission as outcome measure. DESIGN: Naturalistic, prospective, longitudinal. SETTING: Multicenter study in primary care. Health Area 2. Region of Madrid. PARTICIPANTS: Adult patients with a diagnosis of unipolar depression who initiate psychopharmacological treatment with selective serotonin reuptake inhibitor (SSRI) in primary care sites. Patients with history of bipolar disorders, psychotic disorders, dementias, and dependence of toxic substances will be excluded. MAIN MEASUREMENTS: Level of functioning and disability in different domains of well-being assessed through ICF related instruments. Stressful life events, social support and cognitive schemes will be analyzed as mediator variables. Socio-demographic and clinical characteristics, psychopharmacological treatment and treatment compliance are considered independent factors. DISCUSSION AND PRACTICAL USE: Selection bias may affect the generalization of the results. The biopsychosocial model underlying the ICF and its methodology are applied to the study of depression in primary care settings for the first time in Spain. Improving our understanding of disability related factors in depressive patients is expected. This study is one of the main research priorities of the EU (MHADIE project). PMID- 17117337 TI - [Acute tryptophan depletion in eating disorders]. AB - INTRODUCTION: This work describes the rational bases justifying the use of acute tryptophan depletion technique in eating disorders (ED) and the methods and design used in our studies. Tryptophan depletion technique has been described and used in previous studies safely and makes it possible to evaluate the brain serotonin activity. Therefore it is used in the investigation of hypotheses on serotonergic deficiency in eating disorders. Furthermore, and given the relationship of the dysfunctions of serotonin activity with impulsive symptoms, the technique may be useful in biological differentiation of different subtypes, that is restrictive and bulimic, of ED. METHODS: 57 female patients with DSM-IV eating disorders and 20 female controls were investigated with the tryptophan depletion test. A tryptophan-free amino acid solution was administered orally after a two-day low tryptophan diet to patients and controls. Free plasma tryptophan was measured at two and five hours following administration of the drink. Eating and emotional responses were measured with specific scales for five hours following the depletion. A study of the basic characteristics of the personality and impulsivity traits was also done. Relationship of the response to the test with the different clinical subtypes and with the temperamental and impulsive characteristics of the patients was studied. RESULTS: The test was effective in considerably reducing plasma tryptophan in five hours from baseline levels (76%) in the global sample. The test was well tolerated and no severe adverse effects were reported. Two patients withdrew from the test due to gastric intolerance. CONCLUSIONS: The tryptophan depletion test could be of value to study involvement of serotonin deficits in the symptomatology and pathophysiology of eating disorders. PMID- 17117338 TI - [Differential diagnosis of hoarding behaviors]. AB - Hoarding of objects comprises a continuum from normality to extreme disease. It is important to distinguish between the different disorders that include hoarding behaviors. Compulsive hoarding is a form of obsessive-compulsive disorder (OCD) that is characterized by excessive acquisition of possessions, inability to discard possessions, and excessive clutter. Patients usually display other obsessive features, feel distress if they cannot hoard objects, show a typical cognitive pattern with obsessive features, and their interpersonal relations are mediated by objects. Diogenes syndrome is the combination of severe self-neglect, domestic squalor, social withdrawal, hoarding, and refusal of help, in elderly patients. There is high comorbidity with psychiatric/somatic disorders. Depression and dementia are risk factors for self-neglect. Collectionism is a normal phenomenon that is common in children but also found in adults. It is usually an organized activity, and the objects are kept in specific and structured places. The aim of collecting is to organize and hierarchize a series of objects, not just to hoard them. Collected objects are frequently appreciated by other collectors, and become exchanged to enlarge the collection. PMID- 17117339 TI - [Gender differences in cognitive functions and influence of sex hormones]. AB - OBJECTIVE: To review scientific evidence on gender differences in cognitive functions and influence of sex hormones on cognitive performance. METHOD: Systematical search of related studies identified in Medline. RESULTS: Women outperform men on verbal fluency, perceptual speed tasks, fine motor skills, verbal memory and verbal learning. Men outperform women on visuospatial ability, mathematical problem solving and visual memory. No gender differences on attention and working memory are found. Researchers distinguish four methods to investigate hormonal influence on cognitive performance: a) patient with hormonal disorders; b) neuroimaging in individuals during hormone administration; c) in women during different phases of menstrual cycle, and d) in patients receiving hormonal treatment (idiopathic hypogonadotropic hypogonadism, postmenopausal women and transsexuals). The findings mostly suggest an influence of sex hormones on some cognitive functions, but they are not conclusive because of limitations and scarcity of the studies. CONCLUSIONS: There are gender differences on cognitive functions. Sex hormones seem to influence cognitive performance. PMID- 17117340 TI - [Impaired facial emotion recognition in a case of right frontotemporal dementia]. AB - INTRODUCTION: After the description of its involvement in amydalin lesions, there has been growing interest in the last decade on the neuropsychological examination of impaired emotional recognition in different diseases. This study aims to demonstrate the existence of emotional recognition impairment in a case of frontotemporal dementia affecting right temporal lobe structures with an experimental battery. CLINICAL CASE: The case of 7 year long frontotemporal dementia with right temporal predominance, clinically characterized by behavior disorders such as loss of hygiene habits, eating food in bad condition, approach to marginal groups and other psychiatric disorders (megalomanic delusional ideation) is presented. The psychiatric, neurological, neuropsychological and neuroimaging examination are described. METHODS: Facial recognition impairments were assessed with a modification of Ekman and Friesen Task (1976). The results were compared with those obtained in three controls matched by age, and educational level. RESULTS: The case we report showed marked impairment in discrimination, matching, selection and naming of negative facial emotions (anger, fear, sadness and disgust). The impairment was more striking in the selection and naming paradigms. Anger was the most affected emotion. It was hypothesized if the impairment of emotional recognition could be in the base of certain behavior disturbances of the patient such as approach to marginal groups. PMID- 17117343 TI - Cytotoxic and anti-HIV-1 caged xanthones from the resin and fruits of Garcinia hanburyi. AB - Three new caged xanthones, 7-methoxydesoxymorellin (1), 2-isoprenylforbesione (2) and 8,8a-epoxymorellic acid (3), together with nine known caged xanthones were isolated from the EtOAc extracts of resin and fruits of Garcinia hanburyi. The structures were determined by spectroscopic methods. Most of the isolated compounds showed significant cytotoxicities against a panel of mammalian cancer cell lines. Compound 3, together with the known compounds desoxymorellin, morellic acid, gambogic acid, hanburin, forbesione and dihydroisomorellin, exhibited anti-HIV-1 activity in the reverse transcriptase (RT) assay while the known compounds desoxygambogenin and dihydroisomorellin were found moderately active in the syncytium assay. This work represents the first report on the anti HIV-1 activities of caged xanthones. PMID- 17117344 TI - Inhibition of folic acid uptake by catechins and tea extracts in Caco-2 cells. AB - In this present study it was aimed to determine whether the catechins contained in green tea and the whole extracts of Camellia sinensis (Theaceae) inhibit the uptake of folic acid by Caco-2 cell monolayers. Our results indicate that (-) epigallocatechin 3-gallate (EGCG) and (-)-epicatechin 3-gallate (ECG) inhibit cellular folic acid uptake with IC50 values of 34.8 micromol/L and 30.8 micromol/L, respectively. Furthermore, green and black tea extracts were also found to inhibit folic acid uptake with IC50 values of approximately 7.5 and 3.6 mg/mL, respectively. According to these results, simultaneous intake of tea and folic acid may inhibit intestinal folic acid absorption. The consequences with respect to the folate status of the body will need to be examined in vivo. PMID- 17117347 TI - New frontiers in dysphagia rehabilitation. PMID- 17117348 TI - Levels of evidence supporting dysphagia interventions: where are we going? AB - This article describes and compares methodologies for collecting data on the outcomes and efficacy of dysphagia treatments. Several examples are included that distinguish these various types of studies. Study designs that are unique to dysphagia such as assessing the immediate effectiveness of treatment strategies of the instrumental study are described. PMID- 17117349 TI - History and principles of exercise-based therapy: how they inform our current treatment. AB - Exercises designed to strengthen muscles involved in respiration, phonation, and articulation play a key role in the remediation of voice and swallowing disorders. This article presents exercise physiology principles that are beginning to be used by a small group of speech and swallowing researchers to undergird their efficacy-based studies of exercise-based therapy. Three principles--contraction type, task specificity, and overload--are used to compare past exercise-based therapies with present therapies. Comparisons are made between today's methods and Oskar Guttmann's (1893) principles for strengthening muscles of respiration, Emil Froeschels' (1944) therapy to improve laryngeal function, and the myofunctional therapy of the 1960s to improve swallowing and articulation. PMID- 17117350 TI - Respiratory muscle strength training: functional outcomes versus plasticity. AB - Respiratory muscle strength training is a paradigm that has been used for numerous years with a variety of populations including but not limited to spinal cord injury, chronic obstructive pulmonary disease, multiple sclerosis, Parkinson's disease, voice disordered, sedentary elderly, and healthy young. The respiratory muscle strength program discussed here is an expiratory muscle strength training and uses a pressure threshold device with a regimented treatment protocol. The primary purpose of the expiratory muscle strength training program is to promote strength in the expiratory muscles. The training protocol occurs five times per day, 5 days a week, and consists of ~15-20 minutes per day of training by the user at home. The device threshold is changed weekly by a clinician to maintain a threshold load of 75% of an individual's maximum expiratory pressure. The threshold setting of the device is always based on the individual's recorded maximum expiratory pressure generated into a digital pressure gauge. Results of 4 weeks of expiratory muscle strength training protocols indicate up to a 50% improvement for healthy subjects, those with multiple sclerosis, and those with spinal cord injury. The potential transfer of expiratory muscle strength to functional outcomes is discussed, as well as how strength-training paradigms may influence cortical plasticity. PMID- 17117351 TI - Effects of sensorimotor exercise on swallowing outcomes relative to age and age related disease. AB - Parallel to the growing number of adults over age 65 years and the increasing use of exercise in geriatric medicine to improve function and decrease fall risk, recent advances in the treatment of geriatric dysphagia have focused on rehabilitating swallowing function with active exercise. Specific changes in central neural pathways as well as peripheral end organs (muscles) that occur with natural aging may predispose older adults to an increased risk for dysphagia when faced with chronic medical conditions. Research to date primarily has focused on the utility of nonswallow motor exercises to increase muscle strength and range of motion in oropharyngeal structures. Future directions in the field of dysphagia rehabilitation demand evidence-based investigations into the ability of exercise to affect neural plasticity, representing long-lasting alterations in neural organization. PMID- 17117352 TI - Tongue strength and exercise in healthy individuals and in head and neck cancer patients. AB - The tongue plays a critical role in bolus propulsion through the oral cavity and pharynx. This manuscript reviews the types of lingual impairment and overall oropharyngeal swallowing impairment present after treatment for head and neck cancer; specifically, surgery and primary chemoradiotherapy. Oral tongue impairment in surgically treated patients can include reduced range of motion, reduced control, and reduced ability to manipulate, seal, and propel a bolus into the pharynx. Tongue base impairment can result in reduced bolus clearance through the pharynx, resulting in pharyngeal residue and aspiration. The biologic effects of radiotherapy are described, with tissue fibrosis being a primary contributor to development of oropharyngeal swallow disorders. In patients treated with primary chemoradiotherapy, lingual strength has been found to be reduced, as has oral and pharyngeal structural movement during the swallow. The effects of skeletal muscle strengthening programs on muscle physiology are discussed, as are the effects of tongue strengthening exercise programs on tongue strength and swallowing. Future research needs are addressed. PMID- 17117353 TI - What is orofacial fatigue and how does it affect function for swallowing and speech? AB - Speech-language pathologists are likely to encounter patients who report symptoms of fatigue, but there are few clinical procedures to assess this phenomenon. Furthermore, it is difficult to determine whether fatigue contributes to a patient's dysphagia or dysarthria. This article reviews orofacial muscles, including the muscles of the tongue, lips, and cheeks, highlighting in particular their role in swallowing and speaking. It provides definitions of fatigue and describes assessment procedures. The author's research has focused on assessing fatigue, especially of the tongue, and elucidating the effects of exercising the tongue on speech and nonspeech tasks. Most of this work involves people who have Parkinson's disease and neurologically normal adults; results generally support heightened fatigue in Parkinson's disease. However, the effect of fatigue on functional activities remains unclear. Literature regarding the effects of orofacial fatigue on swallowing and speaking is notably sparse, but preliminary evidence indicates that these functions are rather robust. PMID- 17117354 TI - The science and practice of LSVT/LOUD: neural plasticity-principled approach to treating individuals with Parkinson disease and other neurological disorders. AB - Our 15 years of research have generated the first short- and long-term efficacy data for speech treatment (Lee Silverman Voice Treatment; LSVT/LOUD) in Parkinson's disease. We have learned that training the single motor control parameter amplitude (vocal loudness) and recalibration of self-perception of vocal loudness are fundamental elements underlying treatment success. This training requires intensive, high-effort exercise combined with a single, functionally relevant target (loudness) taught across simple to complex speech tasks. We have documented that training vocal loudness results in distributed effects of improved articulation, facial expression, and swallowing. Furthermore, positive effects of LSVT/LOUD have been documented in disorders other than Parkinson's disease (stroke, cerebral palsy). The purpose of this article is to elucidate the potential of a single target in treatment to encourage cross-system improvements across seemingly diverse motor systems and to discuss key elements in mode of delivery of treatment that are consistent with principles of neural plasticity. PMID- 17117355 TI - Clinical implications of cross-system interactions. AB - In this review, we briefly highlight potential cross-system interactions between swallowing and speech production, using data from recent neuroimaging studies, common clinical impairments, cross-system treatment effects, and developmental considerations as supporting evidence. Our overall hypothesis is that speech and swallowing (and other motor behaviors) are regulated through a shared network of brain regions and other neural processes that are modulated on the basis of specific task demands. We emphasize the clinical utility of viewing speech and swallowing as being closely linked from both a diagnostic and treatment perspective. We stress the importance of continuing research to explore the common and perhaps distinct neural circuitry underlying speech and swallowing and the clinical intervention strategies that attempt to capitalize on potential cross-system therapeutic benefits. PMID- 17117356 TI - First report of major histocompatibility complex class II loci from the Amazon pink river dolphin (genus Inia). AB - We report the first major histocompatibility complex (MHC) DQB1 sequences for the two species of pink river dolphins (Inia geoffrensis and Inia boliviensis) inhabiting the Amazon and Orinoco River basins. These sequences were found to be polymorphic within the Inia genus and showed shared homology with cetacean DQB-1 sequences, especially, those of the Monodontidae and Phocoenidae. On the other hand, these sequences were shown to be divergent from those described for other riverine dolphin species, such as Lipotes vexillifer, the Chinese river dolphin. Two main conclusions can be drawn from our results: 1) the Mhc DQB1 sequences seem to evolve more rapidly than other nuclear sequences in cetaceans, and 2) differential positive selective pressures acting on these genes cause concomitant divergent evolutionary histories that derive phylogenetic reconstructions that could be inconsistent with widely accepted intertaxa evolutionary relationships elucidated with other molecular markers subjected to a neutral dynamics. PMID- 17117357 TI - Determination of ancestral proportions in synthetic bovine breeds using commonly employed microsatellite markers. AB - The International Society of Animal Genetics (ISAG) has chosen nine microsatellites (international marker set) as a standard that should be included in all cattle parentage studies. They are BM1824, BM2113, INRA023, SPS115, TGLA122, TGLA126, TGLA227, ETH10, and ETH225. We decided to ascertain whether this microsatellite set could be used to determine ancestral proportions in individual animals of synthetic breeds produced by crossing zebu and taurine cattle. Since the genotypes of these markers are routinely available, this would constitute a practical and cost-free method to estimate the ancestry of synthetic breed animals. Genotypes of 100 Gir and 100 Holstein animals were examined for this ISAG marker set. As expected, there were very significant allele frequency differences between the two breeds at most loci. We also typed 20 Girolando animals for which there was complete genealogical information. "Structure" software easily distinguished Holstein and Gir animals based on their microsatellite genotypes; it also attributed the genomic proportion of zebu and taurine of each of the 20 Girolando animals. The proportion of Holstein ancestry was then regressed on the genealogical data; there was a highly significant correlation (r = 0.84, P < 0.0001). The nine microsatellites that compose the ISAG international marker set were capable of estimating the ancestral Gir and Holstein genomic proportions in individual Girolando animals within narrow confidence limits. This microsatellite set might also be useful for estimating the proportions of taurine and zebu origins in commercial meat products. PMID- 17117358 TI - Evaluation of DNA polymorphisms involving growth hormone relative to growth and carcass characteristics in Brahman steers. AB - Associations of DNA polymorphisms in growth hormone (GH) relative to growth and carcass characteristics in growing Brahman steers (N = 324 from 68 sires) were evaluated. Polymorphisms were an Msp-I RFLP and a leucine/valine SNP in the GH gene as well as a Hinf-I RFLP and a histidine/arginine SNP in transcriptional regulators of the GH gene, Pit-1 and Prop-1. Genotypic frequencies of the GH SNP, Pit-1 RFLP, and Prop-1 SNP were greater than 88% for one of the bi-allelic homozygous genotypes. Genotypic frequencies for the GH Msp-I RFLP genotypes were more evenly distributed with frequencies of 0.43, 0.42, and 0.15 for the genotypes of +/+, +/-, and -/-, respectively. Mixed model analyses of growth and carcass traits with genotype and contemporary group serving as fixed effects and sire fitted as a random effect suggested that sire was a significant source of variation (P < 0.05) in average daily gain, carcass yield, and marbling score. However, measures of growth and carcass traits were similar across GH Msp-I genotypes as steers were slaughtered when fat thickness was estimated to be approximately 1.0 cm. These polymorphisms within the GH gene and/or its transcriptional regulators do not appear to be informative predictors of growth and carcass characteristics in Brahman steers. This is partly due to the high level of homozygosity of genotypes. These findings do not eliminate the potential importance of these polymorphisms as predictors of growth and carcass traits in Bos taurus or Bos taurus x Bos indicus composite cattle. PMID- 17117359 TI - Screening for fragile X syndrome among Brazilian mentally retarded male patients using PCR from buccal cell DNA. AB - Fragile X syndrome is one of the most frequent causes of mental retardation. Since the phenotype in this syndrome is quite variable, clinical diagnosis is not easy and molecular laboratory diagnosis is necessary. Usually DNA from blood cells is used in molecular tests to detect the fragile X mutation which is characterized by an unstable expansion of a CGG repeat in the fragile X mental retardation gene (FMR1). In the present study, blood and buccal cells of 53 mentally retarded patients were molecularly analyzed for FMR1 mutation by PCR. Our data revealed that DNA extraction from buccal cells is a useful noninvasive alternative in the screening of the FMR1 mutation among mentally retarded males. PMID- 17117360 TI - Genetic trend for milk yield in Guzerat herds participating in progeny testing and MOET nucleus schemes. AB - Genetic trends for 305-day milk yield (P305) in Brazilian Guzerat herds under selection were compared. Data from 4898 lactations of 3179 purebred and crossbred cows from various regions of Brazil were used. Milk yield was adjusted for mature age and the contemporary groups were defined as herd and calving year. Genetic parameters were estimated using the MTDFREML program. The model included the random effects of animals and permanent environment, and herd-calving year, calving season and genetic composition as fixed effects. Genetic trends were estimated by linear regression of weighted average estimated breeding values as a function of calving year. The average P305 was 2065 +/- 922 kg and the heritability was 0.23 +/- 0.03. The annual genetic trend in estimated breeding values of cows for P305 was 7.09 +/- 0.71 kg between 1987 and 2004, and 6.47 +/- 2.35 kg between 1997 and 2004. For cows born and raised in the multiple ovulation and embryo transfer (MOET) nucleus, this trend was 36.46 +/- 24.54 kg/year between 1997 and 2004, 183.14 +/- 47.94 kg/year between 1997 and 2000, and 9.13 +/- 19.19 kg/year between 2001 and 2004. An average inbreeding coefficient of 0.04 was found for inbred MOET cows in 2004. Increasing the size of the family and introducing new progenies changed reliabilities and predicted transmitting ability estimates of MOET sires. In conclusion, there was a positive genetic trend for milk yield in the MOET nucleus at low inbreeding coefficients due to the increased accuracy and estimated genetic merit, but no changes in the average milk yield were observed. PMID- 17117361 TI - Expected coalescence times and segregating sites in a model of glacial cycles. AB - The climatic fluctuations of the Quaternary have influenced the distribution of numerous plant and animal species. Several species suffer population reduction and fragmentation, becoming restricted to refugia during glacial periods and expanding again during interglacials. The reduction in population size may reduce the effective population size, mean coalescence time and genetic variation, whereas an increased subdivision may have the opposite effect. To investigate these two opposing forces, we proposed a model in which a panmictic and a structured phase alternate, corresponding to interglacial and glacial periods. From this model, we derived an expression for the expected coalescence time and number of segregating sites for a pair of genes. We observed that increasing the number of demes or the duration of the structured phases causes an increase in coalescence time and expected levels of genetic variation. We compared numerical results with the ones expected for a panmictic population of constant size, and showed that the mean number of segregating sites can be greater in our model even when population size is much smaller in the structured phases. This points to the importance of population structure in the history of species subject to climatic fluctuations, and helps explain the long gene genealogies observed in several organisms. PMID- 17117362 TI - DGAT1 K232A polymorphism in Brazilian cattle breeds. AB - Recent reports identified DGAT1 (EC 2.3.1.20) harboring a lysine to alanine substitution (K232A) as a candidate gene with a strong effect on milk production traits. Our objective was to estimate the frequency of the DGAT1 K232A polymorphism in the main Zebu and Taurine breeds in Brazil as well as in Zebu x Taurine crossbreds as a potential QTL for marker-assisted selection. Samples of 331 animals from the main Brazilian breeds, Nellore, Guzerat, Red Sindhi, Gyr, Holstein, and Gyr x Holstein F1 were genotyped for DGAT1 K232A polymorphism (A and K alleles) using the PCR-RFLP technique. The highest frequency of the A allele was found in the Holstein sample (73%) followed by Gyr x Holstein F1 (39%). Gyr and Red Sindhi showed low frequencies of A alleles (4 and 2.5%, respectively). The A allele was not found in the Nellore and Guzerat samples. Our results could be used to guide association studies between this locus and milk traits in these breeds. PMID- 17117363 TI - Identification of non-specific alkaline phosphatases in hyphal cells of the fungus Neurospora crassa by in situ histochemistry. AB - The present study was designed to identify alkaline phosphatases in non permeabilized hyphal cells of the fungus Neurospora crassa by staining these enzymatic activities with a modified azo dye coupling method. Our strategy allowed the identification of three non-specific alkaline phosphatase activities, one of them possibly being a novel putative enzyme, which is not responsive to either Mg(2+) or EDTA. Another alkaline phosphatase activity, whose location in the hyphal cell is regulated by phosphate, is stimulated by Mg(2+), inhibited by EDTA, and somehow dependent on the expression of the pho-2(+) -encoded Pi repressible alkaline phosphatase. PMID- 17117364 TI - Molecular analysis of apomixis in cassava. AB - Cassava is the main staple for more than 800 million people in the tropics. It is propagated vegetatively by stem cuttings, which maintains superior genotypes but favors disease accumulation and spread. In this report, we present the results of the screening of the progeny and the second generation of the clone UnB 307 for apomixes using microsatellites. A total of 29 plants were screened, representing the maternal plant, its first and second generations that were left to open pollination. About 20% of the offspring were rated as genetically identical plants. This result confirms the facultative apomictic nature of cassava, with high environmental effect. PMID- 17117365 TI - Molecular differentiation in natural populations of Anopheles oswaldoi sensu lato (Diptera: Culicidae) from the Brazilian Amazon, using sequences of the COI gene from mitochondrial DNA. AB - Anopheles (Nyssorhynchus) oswaldoi (Peryassu, 1922) s. l., which has been incriminated as a potential human malaria vector in Western Brazilian Amazon, may constitute a cryptic species complex. However, the most recent study with isozymes indicated high similarity among samples from the States of Acre, Amazonas and Rondonia in the Brazilian Amazon. In the present study, 45 individuals were sequenced from Sena Madureira (State of Acre), Coari (State of Amazonas), Sao Miguel (State of Rondonia), and Moju (State of Para), using the cytochrome oxidase I gene from mitochondrial DNA. Twenty-five haplotypes were identified in the four localities, and no haplotype was shared among them. The lowest haplotype number was detected in the Coari sample. The dendrogram based on maximum parsimony analysis yielded four groups: I) haplotypes 1, 2, 3, 4, and 5 from Sena Madureira and haplotypes 17 and 18 from Sao Miguel; II) haplotypes 13 to 16 and 19 to 22 from Sao Miguel; III) haplotypes 23 to 25 from Moju, and IV) haplotypes 6 to 9 from Sena Madureira and haplotypes 10 to 12 from Coari. The genetic distance (uncorrected p) obtained among the four groups ranged from 0.08 to 5.3%, whereas the highest values (4.97 to 5.3%) were found between groups I (Sena Madureira) and III (Moju). Based on male genitalia identification, it was suggested that group I may be A. oswaldoi s. s. whereas group IV may be A. konderi. Groups II and III could constitute other lineages or species within A. oswaldoi s. l., whose taxonomic status remains to be clarified. These results suggest that additional studies are necessary using samples of A. oswaldoi s. l. from a larger geographic area. PMID- 17117366 TI - Evaluation of the humoral immune response in BALB/c mice immunized with a naked DNA vaccine anti-methicillin-resistant Staphylococcus aureus. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is the major pathogen involved in nosocomial infections, leading to high rates of morbidity and mortality in hospitals worldwide. The methicillin resistance occurs due to the presence of an additional penicillin-binding protein, PBP2a, which has low affinity for beta lactam antibiotics. In the past few years, vancomycin has been the only antibiotic option for treatment of infections caused by multiresistant MRSA; however, reports of vancomycin-resistant strains have generated great concerns regarding the treatment to overcome these infections. In the present study, we report preliminary results regarding the humoral immune response generated in BALB/c mice by two different doses of naked DNA vaccine containing an internal region, comprising the serine-protease domain, of the PBP2a of MRSA. The immunization procedure consisted of four immunizations given intramuscularly within 15-day intervals. Blood was collect weekly and anti-PBP2a-specific antibodies were screened by ELISA. BALB/c mice immunized with DNA vaccine anti PBP2a have shown higher antibody titers mainly after the fourth immunization, and intriguingly, no correlation between the humoral immune response and DNA dose was observed. Our results suggest that the DNA vaccine anti-PBP2a induced an immune response by production of specific antibodies anti-MRSA in a non-dose-dependent manner, and it could represent a new and valuable approach to produce specific antibodies for passive immunization to overcome MRSA infections. PMID- 17117367 TI - Association of BoLA-DRB3.2 alleles with tick (Boophilus microplus) resistance in cattle. AB - Losses caused by bovine tick burdens in tropical countries have a tremendous economic impact on production systems. Besides reducing production, this parasite can cause death in the most susceptible animals. The use of commercial acaricides has been the major method of control, but their misuse has led to tick resistance to many chemicals. More recently, vaccines have been used in some countries without solving the problem completely. An alternative could be the development of resistant animals and the use of genetic markers and candidate genes that could help with the enormous task of selecting resistant animals. The bovine lymphocyte antigen genes (BoLA) have been shown to be associated with some parasitic infestations and disease incidence. Thus, the objective of the present study was to determine the association of BoLA-DRB3.2 alleles with tick resistance in cattle. The study was conducted on 231 F2 (Gyr x Holstein) animals that were artificially infested with 10,000 tick larvae. Log of tick count +1 was used as the dependent variable in a mixed animal model with allele substitution effects in addition to fixed effects of year and season at tick count, sex of calves, age of animal at tick count, hair type (short-straight, short-curl, long straight, and long-curl), coat color (white, >75% white, 50- 75% white, and 25 50% white), and additive genetic, permanent environmental and residual effects as random. Females showed fewer ticks than males. Animals with short-straight hair were more resistant to tick infestation than animals with long-curl hair, and animals with whiter coat color also had fewer ticks. An association between BoLA alleles and lower tick number was found for alleles DRB3.2 *18, *20 and *27 at the 5% significance level. Also, one allele (DRB3.2*16) showed an association at the 10% level. Allele *27 was the most frequent in the population (30.7%), followed by alleles *16 (10.8%), *20 (8.7%) and *18 (2.4%). These results suggest that BoLA-DRB3.2 alleles could be used to help in the selection of animals resistant to tick infestation. However, further studies involving a larger population of cattle in combination with other BoLA genes may help to understand the mechanisms of resistance to parasites. PMID- 17117368 TI - Conservation of glycolytic oscillations in Saccharomyces cerevisiae and human pancreatic beta-cells: a study of metabolic robustness. AB - The present study compares two computer models of the first part of glucose catabolism in different organisms in search of evolutionarily conserved characteristics of the glycolysis cycle and proposes the main parameters that define the stable steady-state or oscillatory behavior of the glycolytic system. It is suggested that in both human pancreatic beta-cells and Saccharomyces cerevisiae there are oscillations that, despite differences in wave form and period of oscillation, share the same robustness strategy: the oscillation is not controlled by only one but by at least two parameters that will have more or less control over the pathway flux depending on the initial state of the system as well as on extra-cellular conditions. This observation leads to two important interpretations: the first is that in both S. cerevisiae and human beta-cells, despite differences in enzyme kinetics and mechanism of feedback control, evolution seems to have kept an oscillatory behavior coupled to the glucose concentration outside the cytoplasm, and the second is that the development of drugs to regulate metabolic dysfunctions in more complex systems may require further study, not only determining which enzyme is controlling the flux of the system but also under which conditions and how its control is maintained by the enzyme or transferred to other enzymes in the pathway as the drug starts acting. PMID- 17117369 TI - The synthesis of a new cassava-derived species, Manihot vieiri Nassar. AB - A new species was synthesized artificially by chromosome doubling in a hybrid. The ensuing polyploid type exhibits an apomictic nature and maintains its morphological characteristics in the progeny. It showed a frequency of multiembryonic sacs of 29% in the ovules examined, whereas sacs were absent in the diploid type. PMID- 17117370 TI - Detection of genetic polymorphism among and within Echinococcus granulosus strains by heteroduplex analysis of a microsatellite from the U1 snRNA genes. AB - Polymerase chain reaction of a pentanucleotide microsatellite in the U1 snRNA gene complex generated a multiple band pattern due to the priming of paralogous sequences. Denaturation and slow renaturation of polymerase chain reaction products allow the formation of heteroduplex DNA that can be detected by its differential mobility in polyacrylamide gel electrophoresis. Heteroduplex analysis was used to determine if the U1 snRNA microsatellite could be a useful genetic marker in Echinococcus granulosus. A U1 snRNA microsatellite fragment from E. granulosus was isolated and characterized by Southern blot and sequencing. Four E. granulosus strains were analyzed: sheep, Tasmanian sheep, cattle, and camel strains. The former two showed polymorphism and shared three of the six patterns found for sheep strain. The cattle strain displayed two patterns, and the camel strain was monomorphic. The electrophoretic profiles were used for statistical analysis in order to determine genetic distance and the relationship among strains. Heteroduplex analysis can be helpful in genotyping E. granulosus strains and is useful in detecting polymorphism within strains. PMID- 17117371 TI - TcZFP8, a novel member of the Trypanosoma cruzi CCHC zinc finger protein family with nuclear localization. AB - In a 17-kb genomic fragment of Trypanosoma cruzi chromosome XX, we identified three tandemly linked genes coding for CX(2)CX(4)HX(4)C zinc finger proteins. We also showed that similar genes are present in T. brucei and Leishmania major, sharing three monophyletic groups among these trypanosomatids. In T. cruzi, TcZFP8 corresponds to a novel gene coding for a protein containing eight zinc finger motifs. Molecular cloning of this gene and heterologous expression as a fusion with a His-tag were performed in Escherichia coli. The purified recombinant protein was used to produce antibody in rabbits. Using Western blot analysis, we observed the presence of this protein in all three forms of the parasite: amastigote, trypomastigote and epimastigote. An analysis of cytoplasmic and nuclear cell extracts showed that this protein is present in nuclear extracts, and indirect immunofluorescence microscopy confirmed the nuclear localization of TcZFP8. Homologues of TcZFP8 in T. brucei are apparently absent, while one candidate in L. major was identified. PMID- 17117372 TI - Quantitative analysis of gemcitabine triphosphate in human peripheral blood mononuclear cells using weak anion-exchange liquid chromatography coupled with tandem mass spectrometry. AB - Gemcitabine triphosphate (dFdCTP) is a highly active metabolite of gemcitabine. It is formed intra-cellularly via the phosphorylation of gemcitabine by deoxycytidine kinase. The monitoring of dFdCTP in human peripheral blood mononuclear cells (PBMCs), in addition to plasma concentrations of gemcitabine and its metabolite 2',2'-difluorodeoxyuridine, is considered very useful in determining pharmacokinetic-pharmacodynamic relationships. We describe a novel sensitive assay for the quantification of dFdCTP in human PBMCs. The method is based on weak anion-exchange liquid chromatography and detection with tandem mass spectrometry (LC-MS/MS). The assay has been validated from 1 ng/ml (lower limit of quantification, LLOQ) to 25 ng/ml (upper limit of quantification, ULOQ) using 180 microl aliquots of PBMC extracts containing approximately 0.648 mg protein or 3.8 x 10(6) lysed PBMCs. The LLOQ is equivalent to 94 fmol/10(6) cells (1 ng/ml = 0.18 ng/180 microl or 0.18 ng/0.648 mg protein = 0.047 ng/10(6) cells or 94 fmol/10(6) cells). This highly sensitive assay is capable of quantifying about 200-fold lower concentrations of dFdCTP in human PBMCs than currently available methods. PMID- 17117373 TI - Multivariable modelling for meta-epidemiological assessment of the association between trial quality and treatment effects estimated in randomized clinical trials. AB - Methodological deficiencies are known to affect the results of randomized trials. There are several components of trial quality, which, when inadequately attended to, may bias the treatment effect under study. The extent of this bias, so far only vaguely known, is currently being investigated by 'meta-epidemiological' re analysis of data collected as part of systematic reviews. As inadequate quality components often co-occur, we maintain that the suspected biases must be evaluated simultaneously. Furthermore, the biases cannot safely be assumed to be homogeneous across systematic reviews. Therefore, a stable multivariable method that allows for heterogeneity is needed for assessing the 'bias coefficients'. We present two general statistical models for analysis of a study of 523 randomized trials from 48 meta-analyses in a random sample of Cochrane reviews: a logistic regression model uses the design of the trials as such to give estimates; a weighted regression model incorporates between-trial variation and thus gives wider confidence intervals, but is computationally lighter and can be used with trials of more general design. In both models, heterogeneity in the bias coefficients can be incorporated in two ways. A stratification approach pools the estimates from models estimated on subgroups of the data. We explore stratification by reviews and by broad trial types, the latter of which gives larger subgroups of the data, circumventing instabilities. A multilevel approach also avoids instabilities and addresses the more fundamental problem of interpretation of the pooled multivariable effect in the presence of heterogeneity. PMID- 17117374 TI - Static SIMS studies of the oxides and hydroxides of aluminium. AB - The degree of hydroxylation or hydration of aluminium surfaces has been examined by static secondary ion mass spectroscopy (SSIMS). The SSIMS spectra of a series of aluminium oxide, oxyhydroxide and hydroxide surfaces have been obtained using instruments in three configurations. Similarities were observed in both negative and positive secondary ions spectra. Even though a direct comparison of the relative intensities cannot be made from one instrument to the other, a similar ranking of the various aluminium hydroxylation states was observed. Several ranking methods are discussed, as well as the similarities and differences observed while using the three instruments. Similar secondary ions were detected whatever the degree of hydroxylation of the aluminium oxide. This argues in favour of the formation of fragments by the combination of individually sputtered atoms or clusters to form the more stable secondaries, rather than the kick-off of 'structure-related' clusters originating directly from the upper surface layer. PMID- 17117375 TI - Multivariate analysis approach to the plasma protein profile of patients with advanced colorectal cancer. AB - The aim of the present study was to identify the pattern of plasma protein species of interest as markers of colorectal cancer (CRC). Using matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS), the plasma protein profile was determined in nine stage IV CRC patients (study group) and nine clean colon healthy subjects (control group). Multivariate analysis methods were employed to identify distinctive disease patterns at protein spectrum. In the study and control groups, cluster analysis (CA) on the complete MALDI-MS spectra plasma protein profile showed a distinction between CRC patients and healthy subjects, thus allowing the identification of the most discriminating ionic species. Principal component analysis (PCA) and linear discriminant analysis (LDA) yielded similar grouping results. LDA with leave-one-out cross validation achieved a correct classification rate of 89% in both the patients and the healthy subjects. PMID- 17117376 TI - Nitrite-assisted peptide iodination and conjugation. AB - In this study, a simple method for selective iodination of peptides and proteins is established. Using angiotensin II as the model system, we demonstrate that nitrite catalyzed the selective iodination of the peptide at the N-terminus in an acidic solution. The N-terminal-iodinated peptides alkylated thiol-containing molecules such as N-acetylcysteine and glutathione to form peptide conjugates in a basic solution. Reactive species formed by increasing the pH of the reaction mixture of sodium nitrite and sodium iodide from 4 to 8 selectively iodinated peptides and proteins at tyrosine and histidine residues. These results show that nitrite is a useful catalyst for peptide and protein ligation. PMID- 17117377 TI - A microfluidic-based system for analysis of single cells based on Ca2+ flux. AB - A microfluidic format-based system has been developed for in situ monitoring of the calcium flux response to agonists using Chinese hamster ovary (CHO) cells. The assay is based on measuring the fluorescent intensity of the calcium sensitive indicator, Fluo-4 AM, and was performed in a modified glass chip channel, whose surface was functionalised using a silanisation method with 3 aminopropyltriethoxysilane (APTS) (enabling the cells to be immobilised on the channel surface). CHO cells calcium flux response was measured for different agonists over a range of concentrations. Cells and reagents were introduced into the chip in a continuous flow as a series of plugs in a given sequence. PMID- 17117378 TI - Integrated isotachophoretic preconcentration with zone electrophoresis separation on a quartz microchip for UV detection of flavonoids. AB - A quartz microchip integrated isotachophoretic (ITP) preconcentration with zone electrophoresis (ZE) separation was fabricated using a novel multi-point pressure method featured in normal temperature and lower pressure during bonding process. ITP followed by subsequential ZE of two flavonoids, quercetin and isorhamnetin on the microchip was performed consecutively on the homemade microfluidic workstation with UV detection, resulting in a decreased detectable concentration of 32-fold, compared to the ZE mode only, and their detection limits decreased down to 0.2 microg/mL and 1.2 microg/mL, respectively. PMID- 17117379 TI - Sacrificial layer microfluidic device fabrication methods. AB - Over the past 15 years, research in the field of microfluidics has experienced rapid growth due to significant potential advantages such as low cost, short analysis times, and elimination of sources of contamination. Although etched and thermally bonded glass substrates have seen widespread use and offer solid performance, device fabrication still remains cumbersome. Recent advances in sacrificial layer microfabrication methods for microfluidics have overcome many disadvantages of conventional fabrication approaches. Phase-changing sacrificial layers have been implemented in making inexpensive and high-performance polymer microchips for electrophoretic analysis, protein focusing, and sample preconcentration. In addition, novel channel fabrication methods based on standard thin-film processes, which are readily integratable with microfabrication techniques used for electrical components, are being applied increasingly for the creation of microfluidic devices. These new sacrificial layer fabrication approaches will be instrumental in making low-cost and high quality polymer microchips, and in interfacing electrical and fluidic systems on glass or semiconductor substrates. PMID- 17117380 TI - Micro free-flow IEF enhanced by active cooling and functionalized gels. AB - Rapid free-flow IEF is achieved in a microfluidic device by separating the electrodes from the focusing region with porous buffer regions. Moving the electrodes outside enables the use of large electric fields without the detrimental effects of bubble formation in the active region. The anode and cathode porous buffer regions, which are formed by acrylamide functionalized with immobilized pH groups, allow ion transport while providing buffering capacity. Thermoelectric cooling mitigates the effects of Joule heating on sample focusing at high field strengths (approximately 500 V/cm). This localized cooling was observed to increase device performance. Rapid focusing of low-molecular-weight p/ markers and Protein G-mouse IgG complexes demonstrate the versatility of the technique. Simulations provide insight into and predict device performance based on a well-defined sample composition. PMID- 17117381 TI - Microfluidics for multiplexed MS analysis. AB - Recent advances of microfluidics systems suitable for multiplexed MS analysis are reviewed with respect to fabrication technologies and applications. PMID- 17117382 TI - Genotyping with microfluidic devices. AB - In the past few years, electrophoresis microchips have been increasingly utilized to interrogate genetic variations in the human and other genomes. Microfluidic devices can be readily applied to speed up existing genotyping protocols, in particular the ones that require electric field-mediated separations in conjunction with restriction fragment analysis, DNA sequencing, hybridization based techniques, allele-specific amplification, heteroduplex analysis, just to list the most important ones. As a result of recent developments, microfabricated electrophoresis devices offer several advantages over conventional slab-gel electrophoresis, such as small sample volume requirement, low reagent consumption, the option of system integration and easy multiplexing. The analysis speed of microchip electrophoresis is significantly higher than that of any other electric field-mediated separation techniques. State-of-the-art microfluidic bioanalytical devices already claim their place in most molecular biology laboratories. This review summarizes the recent developments in microchip electrophoresis methods of nucleic acids, particularly for rapid genotyping, that will most likely play a significant role in the future of clinical diagnostics. PMID- 17117383 TI - Fabrication and characterization of semicircular detection electrodes for contactless conductivity detector - CE microchips. AB - This study uses simple and reliable microfabrication techniques to fabricate CE biochips, integrating a novel contactless conductivity detector in a miniaturized detection system in a microfluidic biochip. The off-channel electrodes are deposited around side channels by Au sputtering and patterned using a standard "lift-off" process. A vacuum fusion bonding process is employed to seal the lower substrate containing the microchannels and the electrodes to an upper glass cover plate. The variations in the capacitance between the semicircular detection electrodes in the side channels are measured as different samples and ions pass through the detection region of the CE separation channel. Samples of Rhodamine B, commercial sports drinks, mineral waters, and a red wine, respectively, are mixed in different buffer solutions, separated, and successfully detected using the developed device. The semicircular detection electrodes for the contactless conductivity detector have microscale dimensions and provide a valuable contribution to the realization of the lab-on-a-chip concept. PMID- 17117384 TI - Floating electrode dielectrophoresis. AB - In practice, dielectrophoresis (DEP) devices are based on micropatterned electrodes. When subjected to applied voltages, the electrodes generate nonuniform electric fields that are necessary for the DEP manipulation of particles. In this study, electrically floating electrodes are used in DEP devices. It is demonstrated that effective DEP forces can be achieved by using floating electrodes. Additionally, DEP forces generated by floating electrodes are different from DEP forces generated by excited electrodes. The floating electrodes' capabilities are explained theoretically by calculating the electric field gradients and demonstrated experimentally by using test-devices. The test devices show that floating electrodes can be used to collect erythrocytes (red blood cells). DEP devices which contain many floating electrodes ought to have fewer connections to external signal sources. Therefore, the use of floating electrodes may considerably facilitate the fabrication and operation of DEP devices. It can also reduce device dimensions. However, the key point is that DEP devices can integrate excited electrodes fabricated by microtechnology processes and floating electrodes fabricated by nanotechnology processes. Such integration is expected to promote the use of DEP devices in the manipulation of nanoparticles. PMID- 17117385 TI - Ionic dispersion in nanofluidics. AB - An analytical solution for dispersion of ionic and neutral solutes in nanoscale channels is presented. Results suggest that in the presence of relatively thick electrical double layers (EDLs) characteristic of nanofluidics, the dispersion of ionic solutes differs from that of neutral solutes on which previous theory is based. Ionic dispersion for circular cross-section channels is quantified as a function of a valance parameter, the relative EDL thickness, and the form of the velocity profile. Two unique mechanisms governing ionic dispersion in both pressure- and electrokinetically driven flows are identified. The results of the analytical solution, employing the linearized form of the Poisson-Boltzmann equation, are supported and extended by the results of an independent computational model employing the nonlinear Poisson-Boltzmann equation. Applicability of the computational results is not limited by the Debye-Huckel approximation. Collectively, these results indicate that dispersion of ionic species in nanoscale channels is markedly charge dependent, and substantially deviates from that of neutral solutes in the same flow. PMID- 17117386 TI - Improvements on the electrokinetic injection technique for microfluidic chips. AB - This paper presents a T-form electrokinetic injection system for the discrete time-based loading and dispensing of samples of variable-volume in a microfluidic chip. A novel push-pull effect is produced during the loading and dispensing processes by the application of an appropriate control voltage distribution. The experimental and numerical results show that this push-pull loading technique produces compact sample plugs and hence improves the detection resolution of the microfluidic device. The injection system is integrated with a microflow switch, and a suitable voltage control scheme is proposed to guide the sample to the desired outlet port such that the microfluidic device can function as a microdispenser. The time-based variable-volume T-form injection method presented in this study is performed using a compact geometry and a simple control scheme and can be readily integrated with other microfluidic devices to form a microfluidic system capable of continuous monitoring and analysis of bioreactions in the life science and biochemistry fields. PMID- 17117387 TI - Numerical analysis of an electrokinetic double-focusing injection technique for microchip CE. AB - The injection techniques in electrophoresis microchips play an important role in the sample-handling process, whose characteristics determine the separation performance achieved, and the shape of a sample plug delivered into the separation channel has a great impact on the high-quality separation performance as well. This paper describes a numerical investigation of different electrokinetic injection techniques to deliver a sample plug within electrophoresis microchips. A novel double-focusing injection system is designed and fabricated, which involves four accessory arm channels in which symmetrical focusing potentials are loaded to form a unique parallel electric field distribution in the intersection of injection channel and separation channel. The parallel electric field effectuates virtual walls to confine the spreading of a sample plug at the intersection and prevents sample leakage into separation channel during the dispensing step. The key features of this technique over other injection techniques are the abilities to generate regular and nondistorted shape of sample plugs and deliver the variable-volume sample plugs by electrokinetic focusing. The detection peak in the proposed injection system is uniform regardless of the position of the detection probe in the separation channel, and the peak resolution is greatly enhanced. Finally, the double-focusing injection technique shows the flexibility in detection position and ensures improved signal sensitivity with good peak resolution due to the delivered high-quality sample plug. PMID- 17117388 TI - Studying drug-plasma protein interactions by two-injector microchip electrophoresis frontal analysis. AB - We developed a simple, rapid, and sensitive two-injector microchip electrophoresis frontal analysis (MCE-FA) method for studying drug-plasma protein interactions. In this method, large volumes of a reference sample and drug-plasma protein mixture were simultaneously introduced into the respective sections of the microchannel through the separated injectors and then electrophoresed. Since the reference sample did not meet with the interacting species during migration, it could be used as an external standard. The interaction between heparin and HSA was quantitatively characterized as a model system. The binding constant was found to be (1.53 +/- 0.01) x 10(4) M(-1). PMID- 17117389 TI - A sheath-flow nanoelectrospray interface of microchip electrophoresis MS for glycoprotein and glycopeptide analysis. AB - Microchip was coupled with MS through a stable, sensitive, and controllable sheath-flow nanoelectrospray (nES) interface for glycoprotein and glycopeptide analysis. The nano-ESI (nESI) was made with a delivery capillary, a commercial nES capillary, and a stainless steel (SS) tube which were connected together through a tee unit. High voltage for nES was applied on the SS tube and the commercial nES capillary was used as nES emitter. The delivery capillary was attached to the microchannel for delivering liquid from microchip to the nESI source. The flow rate of sheath liquid was optimized to be 100-200 nL/min which largely reduced the sample dilution. The detection limit of peptides on this microchip/MS platform was at femtomole level. Glycoprotein and glycopeptides were also successfully analyzed on the platform. All the glycoforms and glycopeptides of ribonuclease B (RNase B) were identified with this method. Some structures of the glycopeptides from RNase B were further characterized with MS/MS on the microchip, coupled with a quadrupole IT-MS. PMID- 17117390 TI - Microchip electrophoresis with wall-jet electrochemical detector: influence of detection potential upon resolution of solutes. AB - This report studies the electrochemical response of wall-jet detector for microchip electrophoresis (microCE). It shows that in wall-jet configuration, the electrochemical detector operates in coulometric mode and that there is an influence of detection potential upon peak width and therefore upon the resolution of solutes. Upon raising the detection potential from +0.3 to +0.9 V, the resolution between model analytes, dopamine and catechol, increases from 0.63 to 2.90. The reasons for this behavior originate in wall-jet detector design and in its typically significant higher detector volume than the volume of injected sample. The conversion efficiency of the wall-jet electrochemical detection cell was found to be 97.4% for dopamine and 98.0% for catechol. The paper brings deeper understanding of operations of wall-jet electrochemical detectors for microchip devices, and it explains previously reported significantly sharper peaks when electrocatalytic electrodes (i.e., palladium and carbon nanotube) were used in microCE-electrochemistry wall-jet detector. PMID- 17117391 TI - A candidate precursor to serous carcinoma that originates in the distal fallopian tube. AB - The tubal fimbria is a common site of origin for early (tubal intraepithelial carcinoma or TIC) serous carcinomas in women with familial BRCA1 or 2 mutations (BRCA+). Somatic p53 tumour suppressor gene mutations in these tumours suggest a pathogenesis involving DNA damage, p53 mutation, and progressive loss of cell cycle control. We recently identified foci of strong p53 immunostaining-termed 'p53 signatures'-in benign tubal mucosa from BRCA+ women. To examine the relationship between p53 signatures and TIC, we compared location (fimbria vs ampulla), cell type (ciliated vs secretory), evidence of DNA damage, and p53 mutation status between the two entities. p53 signatures were equally common in non-neoplastic tubes from BRCA+ women and controls, but more frequently present (53%) and multifocal (67%) in fallopian tubes also containing TIC. Like prior studies of TIC, p53 signatures predominated in the fimbriae (80-100%) and targeted secretory cells (HMFG2 + /p73-), with evidence of DNA damage by co localization of gamma-H2AX. Laser-capture microdissected and polymerase chain reaction-amplified DNA revealed reproducible p53 mutations in eight of 14 fully analysed p53 signatures and all of the 12 TICs; TICs and their associated ovarian carcinomas shared identical mutations. In one case, a contiguous p53 signature and TIC shared the same mutation. Morphological intermediates between the two, with p53 mutations and moderate proliferative activity, were also seen. This is the first report of an early and distinct alteration in non-neoplastic upper genital tract mucosa that fulfils many requirements for a precursor to pelvic serous cancer. The p53 signature and its malignant counterpart (TIC) underline the significance of the fimbria, both as a candidate site for serous carcinogenesis and as a target for future research on the early detection and prevention of this disease. PMID- 17117392 TI - OCT4: biological functions and clinical applications as a marker of germ cell neoplasia. AB - Germ cell tumours (GCTs) are a heterogeneous group of neoplasms, which develop in the gonads as well as in extragonadal sites, that share morphological patterns and an overall good prognosis, owing to their responsiveness to current surgical, chemotherapeutic, and radiotherapeutic measures. GCTs demonstrate extremely interesting biological features because of their close relationships with normal embryonal development as demonstrated by the pluripotentiality of some undifferentiated GCT variants. The similarities between GCTs and normal germ cell development have made it possible to identify possible pathogenetic pathways in neoplastic transformation and progression of GCTs. Genotypic and immunophenotypic profiles of these tumours are also useful in establishing and narrowing the differential diagnosis in cases of suspected GCTs. Recently, OCT4 (also known as OCT3 or POU5F1), a transcription factor that has been recognized as fundamental in the maintenance of pluripotency in embryonic stem cells and primordial germ cells, has been proposed as a useful marker for GCTs that exhibit features of pluripotentiality, specifically seminoma/dysgerminoma/germinoma and embryonal carcinoma. The development of commercially available OCT4-specific antibodies suitable for immunohistochemistry on paraffin-embedded specimens has generated increasing numbers of reports of OCT4 expression in a wide variety of gonadal and extragonadal GCTs. OCT4 immunostaining has been shown to be a sensitive and specific marker for seminomatous/(dys)germinomatous tumours and in embryonal carcinoma variants of non-seminomatous GCTs, whether in primary gonadal or extragonadal sites or in metastatic lesions. Therefore, OCT4 immunohistochemistry is an additional helpful marker both in the differential diagnosis of specific histological subtypes of GCTs and in establishing a germ cell origin for some metastatic tumours of uncertain primary. OCT4 expression has also been reported in pre-invasive conditions such as intratubular germ cell neoplasia, unclassified (IGCNU) and the germ cell component of gonadoblastoma. Additionally, OCT4 immunostaining shows promise as a useful tool in managing patients known to be at high risk for the development of invasive GCTs. PMID- 17117393 TI - Post-stroke dementia and depression: frontosubcortical dysfunction as missing link? AB - OBJECTIVE: Testing the hypothesis that depressive symptoms in dementia reflect dysfunction in fronto-subcortical pathways. BACKGROUND: Both depression and dementia can be the result of vascular damage of the brain. The nature of the depressive symptomatology seems to be related to concommittant cognitive disturbances in that subjects show more so-called motivational symptoms of depression. These symptoms can be the result of frontal-subcortical dysfunction. It could be very helpful for clinical practice if these subjects could be identified by simple diagnostic procedures. METHODS: Associations were computed between measures of depressive symptoms and a set of neuropsychological tests in a sample of 54 subjects with a post-stroke dementia. RESULTS: Although we used an extensive set of neuropsychological tests, most subjects were able to participate only in a small part of them, because of disease severity. Our hypothesis was supported by a negative correlation between scores on the verbal semantic fluency task and the total numbers of motivational depressive symptoms. None of the neuropsychological tests was significantly related to the number of mood symptoms neither did they correlate with the total number of depressive symptoms. CONCLUSION: This study gives further evidence for the assumption that motivational-based depressive symptoms partially originate from fronto subcortical dysfunction. PMID- 17117394 TI - The incidence of first-onset psychotic symptoms and paranoid ideation in a representative population sample followed from age 70-90 years. Relation to mortality and later development of dementia. AB - BACKGROUND: Limited data are available on the incidence of psychotic symptoms in the elderly. OBJECTIVE: To elucidate the incidence of first-onset psychotic symptoms in the elderly and their relation to mortality and later development of dementia. METHOD: A population-sample (n = 392) born 1901-1902 was assessed from age 70-90 with psychiatric examinations, medical record reviews and from age 85, also with key-informant interviews. Individuals developing dementia were excluded. RESULT: The cumulative incidence of first-onset psychotic symptoms was 4.8% (8.0% including key-informant reports in the total sample) and 19.8 % in those who survived to age 85. Sixty-four percent of those with first-onset hallucinations later developed dementia, compared to 30% of those with delusions and 25% of those without psychotic symptoms. CONCLUSIONS: One fifth of non demented elderly who survives up to age 85 develops first-onset psychotic symptoms. Hallucinations predict dementia, but most elderly individuals with first-onset psychotic symptoms do not develop dementia. PMID- 17117395 TI - A double-blind, placebo-controlled trial of memantine in age-associated memory impairment (memantine in AAMI). AB - OBJECTIVES: To determine the safety and efficacy of memantine in treating Age Associated Memory Impairment (AAMI). METHODS: Sixty adults between 50-79 years of age meeting diagnostic requirements for AAMI were randomly assigned to either memantine (titrated to 20 mg) or a matched placebo and treated for 90 days. An extensive battery of computerized cognitive tests was administered at screening, baseline and, thereafter, at monthly intervals. RESULTS AND CONCLUSION: Study results suggest that the primary cognitive effects of memantine in this population are on attention and information processing speed, rather than on memory. There were no differences in adverse events between memantine and placebo. PMID- 17117396 TI - Quantitative structure-activity relationship study of bitter di- and tri-peptides including relationship with angiotensin I-converting enzyme inhibitory activity. AB - Bitterness represents a major challenge in industrial application of food protein hydrolysates or bioactive peptides and is a major factor that controls the flavor of formulated therapeutic products. The aim of this work was to apply quantitative structure-activity relationship modeling as a tool to determine the type and position of amino acids that contribute to bitterness of di- and tri peptides. Datasets of bitter di- and tri-peptides were constructed using values from available literature, followed by modeling using partial least square (PLS) regression based on the three z-scores of 20 coded amino acids. Prediction models were validated using cross-validation and permutation tests. Results showed that a single-component model could explain 52 and 50% of the Y variance (bitterness threshold) of bitter di- and tri-peptides, respectively. Using PLS regression coefficients, it was determined that hydrophobic amino acids at the carboxyl terminus and bulky amino acid residues adjacent to the carboxyl terminal are the major determinants of the intensity of bitterness of di- and tri-peptides. However, there was no significant (p > 0.05) correlation between bitterness of di and tri-peptides and their angiotensin I-converting enzyme-inhibitory properties. PMID- 17117397 TI - Synthesis and characterisation of nonclassical ruthenium hydride complexes containing chelating bidentate and tridentate phosphine ligands. AB - The synthesis and characterisation of nonclassical ruthenium hydride complexes containing bidentate PP and tridentate PCP and PNP pincer-type ligands are described. The mononuclear and dinuclear ruthenium complexes presented have been synthesised in moderate to high yields by the direct hydrogenation route (one-pot synthesis) or in a two-step procedure. In both cases [Ru(cod)(metallyl)(2)] served as a readily available precursor. The influences of the coordination geometry and the ligand framework on the structure, binding, and chemical properties of the M--H(2) fragments were studied by X-ray crystal structure analysis, spectroscopic methods, and reactivity towards N(2), D(2), and deuterated solvents. PMID- 17117398 TI - Efficacy of cognitive rehabilitation in patients with mild cognitive impairment treated with cholinesterase inhibitors. AB - BACKGROUND: Individuals who have Mild Cognitive Impairment (MCI) may be in a transitional stage between aging and Alzheimer's disease (AD). The high rate of conversion from MCI to AD makes early treatment an important clinical issue. Recent evidence suggests that cognitive training intervention may reduce the rate of progression to AD. OBJECTIVES: To evaluate the efficacy of a NeuroPsychological Training (TNP) in patients with MCI who are treated with cholinesterase inhibitors (ChEIs), compared with patients MCI treated only with ChEIs and patients not treated, in a longitudinal, one year follow-up study. METHODS: One year longitudinal and retrospective comparison study of neuropsychological performances in 59 subjects affected by Mild Cognitive Impairment (MCI) according to Petersen's criteria. Fifteen subjects were randomised to receive TNP plus cholinesterase inhibitors; 22 subjects cholinesterase inhibitors alone and 22 subjects no treatment. All the subjects referring memory complaints, corroborated by an informant, underwent a multidimensional assessment concerning neuropsychological, behavioural and functional characteristics, at baseline and after one year follow-up. RESULTS: Subjects without treatment maintained their cognitive, functional and behavioural status after one year; patients treated only with ChEIs improved in depressive symptoms whereas subjects treated with TNP and ChEIs showed significant improvements in different cognitive areas, such as memory, abstract reasoning and in behavioural disturbances, particularly depressive symptoms. CONCLUSIONS: A long-term TNP in ChEIs-treated MCI subjects induces additional cognitive and mood benefits. PMID- 17117399 TI - Stereoselective degradation kinetics of tebuconazole in rabbits. AB - Tebuconazole[(RS)-1-p-chlorophenyl-4,4-dimethyl-3-(1H-1,2,4-triazol-1 ylmethyl)pentan-3-ol] is a potent triazole fungicide and consists of a pair of enantiomers. The enantioselective degradation kinetics of tebuconazole was investigated in rabbits by intravenous (iv) injection. The concentrations of (-) (R)-tebuconazole and (+)-(S)-tebuconazole in plasma and tissues were determined by HPLC with a cellulose tris(3,5-dimethylphenylcarbamate)-based chiral stationary phase. Enantioselective analysis methods for this fungicide in plasma and tissues were developed and validated. Good linearities were obtained over the concentration range of 0.25-25 mg/l for both enantiomers. The degradation followed pseudo-first-order kinetics and the degradation of the (+)-(S) tebuconazole was much faster than that of the (-)-(R)-tebuconazole in plasma after administration of racemic tebuconazole. This study also indicated that environmental assessment of enantiomeric degradation may be needed to fully evaluate risks of tebuconazole use. PMID- 17117400 TI - Enantioselective reduction of ketones with borane catalyzed by tridentate amino alcohols derived from salicylaldehyde. AB - In the presence of tridentate chiral ligand 4e, 99% e.e. value was obtained by asymmetric reduction of ethyl benzoylacetate with borane in toluene at 25 degrees C. Various ligands and reaction conditions were investigated to develop a reasonable mechanism that explains the experimental outcomes. PMID- 17117401 TI - Asymmetric catalysis of Morita-Baylis-Hillman reactions by chiral phosphine Lewis bases bearing multiple phenol groups. AB - In the Morita-Baylis-Hillman (MBH) reactions of arylaldehydes with methyl vinyl ketone, it was observed that in the presence of a catalytic amount of a chiral phosphine Lewis base (CPLB) bearing multiple phenol groups, such as CPLB1 (10 mol %), the corresponding MBH adducts could be obtained in moderate to good yields with low to moderate ee's (4-45% ee) at ambient temperature (10 degrees C) in THF. PMID- 17117402 TI - Synthesis of polymer-type chiral stationary phases and their enantioseparation evaluation by high-performance liquid chromatography. AB - Two new chiral polymers of different molecular weights were synthesized by the copolymerization of (1R,2R)-(+)-1,2-diphenylethylenediamine, phenyl diisocyanate and terephthaloyl chloride. The polymers were immobilized on aminated silica gel to afford two chiral stationary phases. The polymers and the corresponding chiral stationary phases were characterized by Fourier transform-IR, elemental analysis, 1H and 13C NMR. The surface coverages of chiral structural units on the chiral stationary phases were estimated as 0.27 and 0.39 mmol/g, respectively. The enantioseparation ability of these chiral stationary phases was evaluated with a variety of chiral compounds by high-performance liquid chromatography. The effects of the organic additives, the composition of mobile phases, and the injection amount of sample on enantioseparation were investigated. A comparison of enantioseparation ability between these two chiral stationary phases was made. It was believed that the chain length of polymeric chiral selector significantly affected the enantioseparation ability of corresponding chiral stationary phase. PMID- 17117403 TI - Estimation of attributable number of deaths and standard errors from simple and complex sampled cohorts. AB - Estimates of the attributable number of deaths (AD) from all causes can be obtained by first estimating population attributable risk (AR) adjusted for confounding covariates, and then multiplying the AR by the number of deaths determined from vital mortality statistics that occurred in the population for a specific time period. Proportional hazard regression estimates of adjusted relative hazards obtained from mortality follow-up data from a cohort is combined with a joint distribution of risk factor and confounders to compute an adjusted AR. Two estimators of adjusted AR are examined. These estimators differ according to which reference population is used to obtain the joint distribution of risk factor and confounders. Two types of reference populations were considered: (i) the population represented by the baseline cohort and (ii) a population that is external to the cohort. Methods used in survey sampling are applied to obtain estimates of the variance of the AD estimator. These variances can be applied to data that range from simple random samples to multistage stratified cluster samples, which are used in national household surveys. The variance estimation of AD is illustrated in an analysis of excess deaths due to having a non-ideal body mass index using the second National Health and Examination Survey (NHANES) Mortality Study and the 1999-2002 NHANES. These methods can also be used to estimate the attributable number of cause-specific deaths and their standard errors when the time period for the accrual of deaths is short. PMID- 17117404 TI - Nanoparticulate systems for nasal delivery of drugs: a real improvement over simple systems? AB - This review discusses the possible benefits of using nanoparticles for nasal delivery of drugs and vaccines. It considers the various factors affecting particle transport across the nasal tissue. The evidence for the improved transport of drugs, such as peptides and proteins, across the nasal epithelium when formulated in a nanoparticulate system, as compared to an optimal solution formulation, is not convincing. For instance it has been shown that a chitosan solution and especially a chitosan powder formulation was superior in enhancing the nasal absorption of insulin as compared to chitosan nanoparticles. On the other hand, the use of nanoparticles for vaccine delivery seems beneficial in that good immune responses are achieved. This could be due to the fact that small particles can be transported preferentially by the lymphoid tissue of the nasal cavity (NALT). However, apparently no studies have been published comparing directly other adjuvant nasal systems with nanoparticulate systems. PMID- 17117405 TI - Gastric cancers in young and elderly patients show different genomic profiles. AB - Although most gastric cancers occur in elderly patients, a substantial number of cases of this common disease occur in young patients. Gastric cancer is a heterogeneous disease at the genomic level and different patterns of DNA copy number alterations are associated with different clinical behaviour. The aim of the present study was to explore differences in DNA copy number alterations in relation to age of onset of gastric cancer. DNA isolated from 46 paraffin embedded gastric cancer tissue samples from 17 patients less than 50 years of age [median 43 (21-49) years] and 29 patients greater than or equal to 70 years of age [median 75 (70-83) years] was analysed by genome-wide microarray comparative genomic hybridization (array CGH) using an array of 5000 BAC clones. Patterns of DNA copy number aberrations were analysed by hierarchical cluster analysis of the mode-normalized and smoothed log(2) ratios of tumour to normal reference fluorescence signal intensities using TMEV software, after which cluster membership was correlated with age group. In addition, supervised analysis was performed using CGH Multi-array. Hierarchical cluster analysis of the array CGH data revealed three clusters with different genomic profiles that correlated significantly with age (p = 0.006). Cluster 1 mainly contained young patients, while elderly patients were divided over clusters 2 and 3. Chromosome regions 11q23.3 and 19p13.3 contributed most to age-related differences in tumour profiles. Gastric cancers of young and old patients belong to groups with different genomic profiles, which likely reflect different pathogenic mechanisms of the disease. PMID- 17117406 TI - Eye size in geckos: Asymmetry, allometry, sexual dimorphism, and behavioral correlates. AB - The function of the vertebrate eye depends on its absolute size, and the size is presumably adapted to specific needs. We studied the variation of eye size at all levels, from intra-individual to inter-specific, in lid- less, spectacled, gecko lizards (Gekkonomorpha). We mea sured 1,408 museum specimens of 62 species, representing subfamilies Diplodactylinae, Gekkoninae, and Sphaerodactylinae. Intra-individually, eye size showed significant directional asymmetry in Stenodactylus sthenodactylus. A latitudinal study of six species confirmed that during postnatal ontogeny eye size undergoes conventional negative allometry; the slope is steeper among adults than among juveniles, expressing the need of juveniles for relatively larger eyes. Within species with sexual size dimorphism, commonly the larger sex possessed larger eyes in absolute terms but not relative to head-and-body length. Interspecifically, eye size showed negative allometry, with slope significantly steeper than those of intraspecific ontogenetic allometry, again expressing the need of juveniles for relatively larger eyes and showing that eye-size differences among species do not merely result from body size differences. Finally, adult eye size varied interspecifically in correlation with parameters of behavioral ecology: eyes were significantly larger in nocturnal than in diurnal species, and significantly larger in cursorial than in scansorial species. PMID- 17117407 TI - Novel mutations in the small leucine-rich repeat protein/proteoglycan (SLRP) genes in high myopia. AB - The importance of the genetic component in high myopia has been well established in population and family studies, but only a few candidate genes have been explored to date. The extracellular matrix small leucine-rich repeat proteins/proteoglycans (SLRPs) regulate collagen fibril diameter and spacing. Given their role in extracellular matrix assembly and expression in the eye, they are likely to regulate its shape and size. Analysis of 85 English and 40 Finnish subjects with high myopia (refractive error of -6 diopters [D] or greater) resulted in 23 sequence variations in four SLRP genes, LUM, FMOD, PRELP, and OPTC. We observed higher number of variations in OPTC in English patients than in controls (p=0.042), and a possibly protective variation in LUM (c.893-105G>A) with p-value of 0.0043. Two intronic variations, six nonsynonymous and one synonymous amino acid changes, were not found in any of the nonmyopic controls. Five changes were detected in opticin, Thr177Arg, Arg229His, Arg325Trp, Gly329Ser, and Arg330His, and all but one (Arg229His) were shown to cosegregate with high myopia in families with incomplete penetrance. A homology model for opticin revealed that Arg229His and Arg325Trp are likely to disrupt the protein structure, and PolyPhen analysis suggested that Thr177Arg, Arg325Trp, and Gly329Ser changes may be damaging. A Leu199Pro change in lumican and Gly147Asp and Arg324Thr variations in fibromodulin are located in the highly conserved leucine-rich repeat (LRR) domains. This study provides new insight into the genetics of high myopia, suggesting that sequence variations in the SLRP genes expressed in the eye may be among the genetic risk factors underlying the pathogenesis of high myopia. PMID- 17117408 TI - Identification of N-terminal modification for recombinant monoclonal antibody light chain using partial reduction and quadrupole time-of-flight mass spectrometry. AB - Since most recombinant monoclonal antibodies (mAbs) contain glutamic acid or glutamate at their N-terminus, cyclization of these residues to form pyroglutamate is an important degradation pathway that often occurs in therapeutic mAb development. In this work, a rapid method was developed to determine pyroglutamate at the N-terminus of mAb light chain by liquid chromatography coupled with electrospray ionization on a quadrupole time-of flight mass spectrometer (QTOF). High levels of pyroglutamate were found at the N terminus of the light chain of a typical recombinant mAb. The quantitative results were comparable to those obtained with a more conventional peptide mapping method. The direct method outlined here can be used to evaluate the impact of N-terminal cyclization during the processing of recombinant mAbs. PMID- 17117409 TI - Lyophilization cycle development for a high-concentration monoclonal antibody formulation lacking a crystalline bulking agent. AB - An efficient freeze-dry cycle was developed for a high concentration monoclonal antibody formulation lacking a crystalline bulking agent. The formulation, at multiple protein concentrations, was characterized using differential scanning calorimetry (DSC) and freeze-dry microscopy. At low protein concentrations the glass transition temperature of the maximally freeze-concentrated solution (T(g)') determined by DSC was similar to the collapse temperature determined by freeze-dry microscopy. However, at higher protein concentrations, the difference between collapse temperature and T(g)' became progressively larger. The difference between the onset temperature for collapse and the complete collapse temperature also became progressively larger as protein concentration increased. JMP Design of Experiment studies were used to assess the effect of freezing rate, primary drying shelf temperature, and chamber pressure on primary drying product temperature, length of primary drying, and product quality attributes. Primary drying was shortened significantly by adjusting to conditions where the product temperature substantially exceeded T(g)' without any apparent detrimental effect to the product. PMID- 17117410 TI - Myasthenia gravis associated with HTLV-I infection and atypical brain lesions. AB - We report a patient who experienced progressive diplopia and distal weakness of the upper limbs. Magnetic resonance imaging of the brain showed extensive white matter lesions and analysis of cerebrospinal fluid revealed acute human T lymphotropic virus type I (HTLV-I) infection. Myasthenia gravis (MG) was evidenced by electromyography (EMG) and antibodies against acetylcholine receptor. This unusual case of MG associated with HTLV-I infection and brain restricted lesions underscores the possible link between viruses and MG pathogenesis. PMID- 17117411 TI - Posterior root-muscle reflexes elicited by transcutaneous stimulation of the human lumbosacral cord. AB - Continuous epidural stimulation of lumbar posterior root afferents can modify the activity of lumbar cord networks and motoneurons, resulting in suppression of spasticity or elicitation of locomotor-like movements in spinal cord-injured people. The aim of the present study was to demonstrate that posterior root afferents can also be depolarized by transcutaneous stimulation with moderate stimulus intensities. In healthy subjects, single stimuli applied through surface electrodes placed over the T11-T12 vertebrae with a mean intensity of 28.6 V elicited simultaneous, bilateral monosynaptic reflexes in quadriceps, hamstrings, tibialis anterior, and triceps surae by depolarization of lumbosacral posterior root fibers. The nature of these posterior root-muscle reflexes was demonstrated by the duration of the refractory period, and by modifying the responses with vibration and active and passive movements. Stimulation over the L4-L5 vertebrae selectively depolarized posterior root fibers or additionally activated anterior root fibers within the cauda equina depending on stimulus intensity. Transcutaneous posterior root stimulation with single pulses allows neurophysiological studies of state- and task-dependent modulations of monosynaptic reflexes at multiple segmental levels. Continuous transcutaneous posterior root stimulation represents a novel, non-invasive, neuromodulative approach for individuals with different neurological disorders. PMID- 17117412 TI - Role of GABA in anxiety and depression. AB - This review assesses the parallel data on the role of gamma-aminobutyric acid (GABA) in depression and anxiety. We review historical and new data from both animal and human experimentation which have helped define the key role for this transmitter in both these mental pathologies. By exploring the overlap in these conditions in terms of GABAergic neurochemistry, neurogenetics, brain circuitry, and pharmacology, we develop a theory that the two conditions are intrinsically interrelated. The role of GABAergic agents in demonstrating this interrelationship and in pointing the way to future research is discussed. PMID- 17117413 TI - Primary colorectal tumors fail to express the proapoptotic mediator PTAG and its reexpression augments drug-induced apoptosis. AB - Genes implicated in tumor evolution and progression, including those in apoptotic pathways, are associated with methylation-associated gene silencing in different tumor types. By exploiting differential methylation we recently isolated a novel pituitary tumor derived apoptosis gene (PTAG) that augments drug-induced apoptosis. The importance of PTAG was determined in other tumor types, and these studies show that the majority of primary colorectal tumors fail to express the PTAG gene, indicating an important role for PTAG in colorectal tumorigenesis. The effects of expression of PTAG were examined through stable transfection of the colorectal cell lines HCT116 and SW480. Expression of PTAG, per se, had no discernible effects on cell viability or cell kinetics. In contrast to these findings, in cells subject to drug challenges that engaged either a death receptor mediated or mitochondrial pathway, all of the experiments indicated a role for PTAG in the intrinsic pathway of apoptosis. Loss of PTAG therefore contributes to a blunted apoptotic response and is likely to predispose cells toward malignant transformation and resistance to chemotherapeutic interventions. PMID- 17117414 TI - The (epi)genetics of human synovial sarcoma. AB - Human synovial sarcomas are aggressive soft tissue tumors with relatively high rates of recurrences and metastases. They display a variable response to common treatment protocols such as radiation and chemotherapy. For the development of novel diagnostic, prognostic, and therapeutic approaches, detailed information on the molecular mechanisms underlying the development of these tumors is of imperative importance. Fusion of the SS18 and (one of the) SSX genes is a molecular hallmark of human synovial sarcomas. The SS18 and SSX genes encode nuclear proteins that exhibit opposite transcription regulatory activities, likely through epigenetic mechanisms. The SS18 protein functions as a transcriptional coactivator and interacts directly with members of the epigenetic chromatin remodeling and modification machineries. In contrast, the SSX proteins function as transcriptional corepressors and are associated with several Polycomb group proteins. Since the domains involved in these apparently opposite transcription regulatory activities are retained in the SS18-SSX fusion proteins, we hypothesize that these fusion proteins function as "activator-repressors" of transcription. The implications of this model for human synovial sarcoma development and future treatment are discussed. PMID- 17117415 TI - Characterization of the native CREB3L2 transcription factor and the FUS/CREB3L2 chimera. AB - CREB3L2 was first identified as the 3'-partner of FUS in a fusion gene that seems to be specific for low grade fibromyxoid sarcoma. In silico analyses suggest that the predicted CREB3L2 protein is a member of the CREB3 family of transcription factors, with its bZIP domain being highly similar to that in CREB3L1, CREB3L3, CREB3L4, CREB3, and Drosophila Bbf-2. In the present study, the authors assessed various cellular outcomes after transfection of NIH3T3 and HEK-293 cells with constructs containing full-length and truncated versions of CREB3L2 and FUS/CREB3L2. Northern blot of CREB3L2 mRNA revealed a 7.4 kbp band that contains 0.4 kbp and 5.5 kbp untranslated 5' and 3' regions, respectively. CREB3L2 constructs containing the first 120 amino acids (aa) showed the highest transcriptional activation. Much stronger transcriptional activation was consistently seen for the FUS/CREB3L2 constructs than for the corresponding CREB3L2 constructs. Transcriptional activity was achieved through the box-B element, ATF6 and CRE binding sites, as well as the GRP78 promoter. Proteins encoded by full-length CREB3L2 and FUS/CREB3L2 were localized to reticular structures of the cytoplasm, whereas the corresponding, truncated proteins lacking the transmembrane domain and the carboxy-terminal part of CREB3L2 resided within the nucleus. The results of the present study show that CREB3L2 is not only structurally, but also functionally very similar to CREB3L1. Thus, studies regarding the pathways influenced by wild-type CREB3L2 should provide valuable clues to the pathogenetic significance of the FUS/CREB3L2 chimera in low grade fibromyxoid sarcoma. PMID- 17117416 TI - Imatinib mesylate induces hypophosphatemia in patients with chronic myeloid leukemia in late chronic phase, and this effect is associated with response. AB - Imatinib is the drug of first choice for most patients with chronic phase chronic myeloid leukemia (CML). Although it is generally well tolerated, a number of hematological and nonhematological side-effects have been described. We report here that imatinib induces hypophosphatemia in a high proportion of our series of CML patients previously treated with interferon alpha, and that this previously unreported side effect is associated with response. PMID- 17117417 TI - Novel microdeletion in the transforming growth factor beta type II receptor gene is associated with giant and large cell variants of nonsmall cell lung carcinoma. AB - Mutations in the tumor suppressor gene transforming growth factor beta (TGFB) Type II receptor (TGFBR2) are frequently found in many cancers with microsatellite instability, but are less common in lung cancer. In the present study, we looked for mutations in TGFBR2 in nonsmall cell lung carcinoma (NSCLC) cells and tissues. A novel homozygous microdeletion (c.492_507del) was identified in two cell lines derived from the same giant cell carcinoma (GCC) and was confirmed in the corresponding tumor tissues. Furthermore, a heterozygous c.492_507del was found in the germ-line of one patient, as well as in the other GCC cases and some large cell carcinomas (LCC) but not in other subtypes of NSCLC. The 16 bp-microdeletion introduced a premature stop codon at positions 590 592 of the cDNA, resulting in a truncated TGFBR2 protein with a mutated transmembrane domain and loss of kinase domain. The GCC cells were characterized as being unresponsive to TGFB induction both in growth inhibition and stimulation of extracellular matrix protein. Moreover, after the reconstitution of wild-type TGFBR2 expression, the sensitivity to TGFB was restored. Therefore, mutated TGFBR2 seems to play an important role in the abrogation of TGFB signal transduction in GCC cells. PMID- 17117418 TI - In vivo and in vitro measurement of brain phosphodiesterase 4 in rats after antidepressant administration. AB - Based largely on in vitro measurements, the mechanism of several antidepressant treatments is thought to involve upregulation of 3'-5'-cyclic adenosine monophosphate (cAMP) signal transduction cascade and a corresponding increase in phosphodiesterase (PDE) 4, the enzyme that metabolizes cAMP. To assess the in vivo status of PDE4, rats were chronically treated with imipramine and then studied with: (1) in vivo positron emission tomography (PET) measurement of (R) [(11)C]rolipram binding, (2) in vitro measurement of [(3)H]rolipram binding in brain homogenates, and (3) Western blotting for protein levels of PDE4 isoforms. Imipramine administration caused no significant change in B(max)/K(d), for both in vivo measurements with (R)-[(11)C]rolipram and in vitro measurements with [(3)H]rolipram in frontal cortex, hippocampus, and diencephalon. None of 10 isoforms of PDE4A, B, and D measured with immunoblots of frontal cortex and hippocampus showed a significant change. In summary, using relatively large brain regions for both in vivo imaging and in vitro measures of radiolabeled ligand binding and protein levels, chronic imipramine treatment via continuous mini-pump administration caused no significant change in PDE4 levels. Most, but not all, prior in vitro studies have found increased PDE4 levels after antidepressant administration. The current results raise questions about the in vivo effects of antidepressant treatment on PDE4 and on other potentially important experimental factors (e.g., continuous infusion vs. intermittent injection of antidepressant) in large brain areas. However, the results do not deny possibility of changes in discrete areas, which were not studied in the current study applying PET. PMID- 17117419 TI - Topographical organization of GABAergic neurons within the ventral tegmental area of the rat. AB - The ventral tegmental area (VTA), the origin of dopaminergic cell bodies that comprise the mesocorticolimibic DA system, is widely implicated in drug and natural reward, cognition, and several psychiatric disorders. In addition to dopaminergic neurons, this region is populated by GABAergic neurons, which both regulate the firing of their dopaminergic counterparts and send projections throughout the brain. Although the dopaminergic neurons of the VTA have been extensively characterized neuroanatomically, much less is known about the GABAergic neurons in this region. Recent data suggest that the rostro-caudal topographic organization of these GABAergic neurons may correspond to their ability to regulate drug reward. In the present study, we used immunohistochemical techniques to examine the frequency and topography of GABAergic neurons throughout the rostro-caudal axis of the VTA and the extent to which they coexpress other proteins, including tyrosine hydroxylase (a marker of DA neurons), cholecystokinin, parvalbumin, calretinin, and calbindin d 28k. PMID- 17117420 TI - Analysis of phosphorylation sites on focal adhesion kinase using nanospray liquid chromatography/multiple reaction monitoring mass spectrometry. AB - An approach based on nanospray liquid chromatography/multiple reaction monitoring mass spectrometry (LC/MRM-MS) was developed in order to analyze twenty-nine phosphorylated and non-phosphorylated tryptic peptides from focal adhesion kinase (FAK). All peptides monitored were resolved and showed excellent peak shape with the exception of one doubly phosphorylated peptide. Optimization of the LC method enabled the identification and subsequent monitoring of six important tyrosine phosphorylation sites on FAK, including phosphorylated Y397 (pY397), pY407, pY576, pY577, pY861, and pY925. This technique was able to identify sites of phosphorylation on FAK as well as qualitatively differentiate between autocatalytic and Src-induced phosphorylation events. FAK was shown to have autocatalytic function, which resulted in efficient phosphorylation of Y397. FAK was also capable of autophosphorylation on residues Y407 and Y576, though apparently less effectively than autophosphorylation at Y397. Src was found to phosphorylate FAK at Y407, Y576, Y577, and Y861. The presence of Src increased the abundance of pY576 at low temperature indicating Src had particularly high kinase activity toward this residue. Furthermore, Src phosphorylated FAK at Y577 to produce FAK bis-phosphorylated at Y576 and Y577. In addition, six novel sites of phosphorylation (Y148, Y347, Y441, T503, S850, and Y1007) were identified on FAK. Interestingly, Src phosphorylated FAK to form a peptide uniquely phosphorylated on Y407, together with substantial amounts of the bis phosphorylated pY397pY407 peptide. These findings will impact significantly on future studies of FAK activity since pY397 is often used as a measure of FAK activity and Src association. PMID- 17117421 TI - Different levodopa actions on the extracellular dopamine pools in the rat striatum. AB - Levodopa has been the mainstay treatment for Parkinson's disease for several decades, but the precise mechanism for its therapeutic action is still not well understood. To date, little distinction has been made between the effects of levodopa on the different brain DA pools. We studied the levodopa action on two extracellular DA pools: one was analyzed by microdialysis (often considered as indicative of volume transmission) and the other by in vivo amperometry during nigrostriatal cell stimulation (more indicative of neurotransmission). Levodopa administration induced a moderate (increased 200%) and tardy (began at 60 min) increase in the DA-pool measured by microdialysis, an effect that increased (increased 500%) and accelerated (began at 10 min) after DA-cell degeneration. Levodopa action on the DA-pool measured by amperometry was very fast (10 min) and prominent (increased 600%) in normal rats. The DA-denervated striatum showed a fast exhaustion during cell stimulation, which prevented further study of the levodopa effect on the DA amperometry-pool under this condition. This study suggests a different kinetic for levodopa action on the volume transmitter and neurotransmitter DA-pool, showing marked changes in levodopa action in the denervated striatum. PMID- 17117422 TI - Effect of a P-glycoprotein inhibitor, Cyclosporin A, on the disposition in rodent brain and blood of the 5-HT1A receptor radioligand, [11C](R)-(-)-RWAY. AB - Limited brain uptake of radioligands with otherwise optimal properties for imaging brain receptors can be improved by blocking the effect of P-glycoprotein (P-gp), an efflux pump at the blood-brain barrier. Using small animal positron emission tomography (PET), we investigated how P-gp and its blockade with Cyclosporin A (CsA) affect rodent brain uptake of [(11)C](R)-(-)-RWAY, a radioligand for brain 5-HT(1A) receptors. Brain uptake of radioactivity was compared in control and CsA-treated rats as well as P-gp knockout and wild type mice. Parent radioligand and radiometabolite in plasma and brain samples were determined at 30 min after injection of radioligand. PET binding potential (BP) was calculated with a reference tissue model. P-gp knockout mice had 2.5- and 2.8 fold greater brain uptake of [(11)C](R)-(-)-RWAY than wild type ones, measured by in vivo PET and ex vivo tissue sampling, respectively. Similarly, CsA increased rat brain uptake 4.9- and 5.3-fold, in vivo and ex vivo. In addition, CsA increased the plasma free fraction of [(11)C](R)-(-)-RWAY in rats by 2.7-fold. Although CsA increased brain uptake in wild type mice by 2.5-fold, it had no effect on plasma free fraction in knockout animals. Three radiometabolites of [(11)C](R)-(-)-RWAY were uniformly distributed in rat brain, suggesting they were inactive. CsA treatment increased brain uptake of [(11)C](R)-(-)-RWAY and only one of its radiometabolites. Regional rat brain BP increased 27-70% in the CsA treated rats and the P-gp knockout mice. [(11)C](R)-(-)-RWAY is a P-gp substrate in rat and mouse. The effects of CsA in rats are mediated by both P-gp blockade and displacement of the radiotracer from plasma proteins. Studies with wild type and knockout mice showed that CsA affected only P-gp in this species. PMID- 17117423 TI - Amphetamine-induced dopamine release: duration of action as assessed with the D2/3 receptor agonist radiotracer (-)-N-[(11)C]propyl-norapomorphine ([11C]NPA) in an anesthetized nonhuman primate. PMID- 17117424 TI - Regulation of morphine reward and feeding by CREB in the lateral hypothalamus. PMID- 17117425 TI - Acute and repeated administration of fluoxetine, citalopram, and paroxetine significantly alters the activity of midbrain dopamine neurons in rats: an in vivo electrophysiological study. AB - We examined the effect of the administration of the selective serotonin reuptake inhibitors (SSRIs) fluoxetine, citalopram, and paroxetine on the activity of spontaneously active dopamine (DA) neurons in the substantia nigra pars compacta (SNC) and ventral tegmental area (VTA) in anesthetized adult male Sprague-Dawley rats. This was accomplished using the technique of in vivo extracellular recording. A single injection of 2.5 mg/kg (i.p.) of fluoxetine significantly increased the number of spontaneously active SNC and VTA DA neurons. In contrast, a single injection of either 1 mg/kg (i.p.) of paroxetine or 5 mg/kg of fluoxetine significantly increased the number of spontaneously active VTA DA neurons. The repeated administration (one injection per day for 21 days) of all of the SSRIs produced a significant increase in the number of spontaneously active VTA DA neurons. Overall, our results indicate that the systemic administration of SSRI alters the activity of midbrain DA neurons with differential effects on VTA compared with SNC DA neurons. PMID- 17117426 TI - Lysosomal sequestration of amine-containing drugs: analysis and therapeutic implications. AB - Amine-containing drugs represent a very important class of therapeutic agents, with the majority of all drugs containing at least one basic nitrogen. For many decades, it has been known that weakly basic compounds can be sequestered into acidic organelles such as lysosomes. Some amines can achieve very high concentrations and induce a dramatic expansion (vacuolization) of the compartment. In the early 70s, Nobel laureate and discoverer of lysosomes, Christian de Duve et al. wrote an elegant commentary describing the theoretical basis for lysosomal sequestration of amines, referring to the process as pH partitioning and the substrates as lysosomotropics. Recently, a resurgence of interest in the intracellular distribution of drugs has occurred considering its therapeutic importance. Specifically, lysosomal sequestration of amines has received considerable attention for reasons including its involvement in drug resistance, inducement of phospholipidosis, and its influence on whole body distribution/pharmacokinetics. Moreover, the sequestration phenomenon has been recently exploited in the development of a novel drug targeting strategy. This review will focus on these occurrences/developments and conclude with a commentary on the expected impact that knowledge regarding the intracellular distribution of drugs will likely have on future drug development processes. PMID- 17117427 TI - Optimization of fed-batch production of the model recombinant protein GFP in Lactococcus lactis. AB - Optimization of recombinant protein production using lactic acid bacteria (LAB) remains an important obstacle on the road to realizing LAB as oral vaccine delivery vehicles. Despite this, there have been few published investigations to explore the higher limits of LAB recombinant protein expression in fed-batch fermentations. In this study, results from response surface experiments suggested an optimal set of conditions for expression of green fluorescent protein (GFP), a model recombinant protein, in bench-scale, fed-batch Lactococcus lactis IL1403 fermentations. The 48 4-L fed-batch fermentations in this set of experiments, along with preliminary studies, investigated the effects of pH, temperature, hemin concentration, concentration of the nisin inducer per cell, and time of induction. Cell densities in this data set ranged from 2.9 to 7.4 g/L and maximum GFP expression per cell ranged from 0.1 to 4.4 relative fluorescence units (RFU)/g. The optimal 4-L, fed-batch fermentation process found here yields growth and protein expression values that dramatically improve upon results from traditional test tube and flask processes. Relative to the traditional process, the experimental optimum conditions yield 4.9 times the cell density, 1.6 times the protein per cell mass, and 8 times the total protein concentration. Unexpectedly, experiments also revealed that the compound hemin, known previously to improve growth and survival of Lactococcus lactis (L. lactis), negatively impacted recombinant protein production when added in concentrations from 5 to 20 microg/mL with this strain. The improvement in protein expression over traditional processes demonstrated here is an important step toward commercial development of LAB for oral delivery of recombinant vaccines and therapeutic proteins. PMID- 17117428 TI - Polymorphism and solvatomorphism 2005. AB - Papers and patents that deal with polymorphism (crystal systems for which a substance can exist in structures characterized by different unit cells, but where each of the forms consists of exactly the same elemental composition) and solvatomorphism (systems where the crystal structures of the substance are defined by different unit cells, but where these unit cells differ in their elemental composition through the inclusion of one or molecules of solvent) have been summarized in an annual review. The works cited in this review were published during 2005, and were drawn primarily from the major physical, crystallographic, and pharmaceutical journals. The review is divided into sections that cover articles of general interest, computational and theoretical studies, preparative and isolation methods, structural characterization and properties of polymorphic and solvatomorphic systems, studies of phase transformations, effects associated with secondary processing, and United States patents issued during 2005. PMID- 17117429 TI - The connection between the steady state (Vss) and terminal (Vbeta) volumes of distribution in linear pharmacokinetics and the general proof that Vbeta >/= Vss. AB - The steady state and terminal (area) volumes of distribution are important pharmacokinetic parameters defined as the ratio of the total quantity of drug in the body, A(b)(t), to drug plasma concentration C(p)(t) at steady state and the terminal phase of drug elimination, respectively. The general equations for the approach of C(p)(t), A(b)(t) and the distribution volume A(b)(t)/C(p)(t) to the steady state values (for a continuous constant rate drug infusion) are derived. It is shown that the time course of A(b)(t) near the asymptotic steady state value depends on both the terminal and steady state volumes of distribution, and an accurate equation to determine the time required to reach the steady state is obtained. For a general linear pharmacokinetic system (i.e., with possible drug elimination at any state from any compartment and drug exchange between compartments) it is proven that V(beta) >/= V(ss). A physiologically determined feature, which is the drug input into plasma for reaching the steady state or terminal phase, underlies the proof. If the steady state is reached by a continuous input of drug into some compartment other than plasma, and the terminal volume of distribution is considered after dosing of a drug in the same compartment, then both cases V(ss) <> V(beta) are possible. It is shown that the general exponential series for C(p)(t) after intravenous bolus dose may have negative pre-exponents, unlike a common assumption that all pre-exponents should be positive. Its is figured out that the commonly used equations for the estimation of V(ss) and V(beta) (V(ss) = D x AUMC/AUC(2) and V(beta) = D/(AUC x beta) may yield V(ss) > V(beta) for a linear pharmacokinetic system, contrary to the usual statement (V(ss) < V(beta)) and its seemingly simple proof, which has a flaw. It is shown that the time required to reach the steady state concentration of drug in plasma could be much shorter than a commonly used estimation of 5t(1/2), where t(1/2) is the terminal half-life obtained from the intravenous bolus drug plasma concentration time course. PMID- 17117430 TI - Impact of renal dysfunction on operative mortality following endovascular abdominal aortic aneurysm surgery. AB - BACKGROUND: Preoperative renal dysfunction is a significant risk factor for death after open abdominal aortic aneurysm repair. The aim of this study was to determine whether renal dysfunction also affected mortality after endovascular aneurysm repair. METHODS: Patients from the EUROSTAR registry were stratified into two groups: 4198 with normal renal function (creatinine less than 133 micromol/ml) and 969 with renal dysfunction (serum creatinine more than 133 micromol/ml). Patient characteristics and postoperative complications in the two groups were compared and the effect of renal dysfunction on operative mortality was analysed by multivariable regression models. RESULTS: Patients with renal dysfunction had significantly more co-morbidities, including cardiac and pulmonary impairment. Thirty-day mortality was significantly higher in the group with renal dysfunction (6.2 versus 2.0 per cent; P<0.001). A significant increase in mortality (5.5 per cent) was also seen in patients with moderate renal dysfunction (serum creatinine 133-265 micromol/ml). After adjustment for age and other risk factors, renal dysfunction was still an independent risk factor for 30 day mortality (odds ratio 2.3, 95 per cent confidence interval 1.6 to 3.3; P<0.001). CONCLUSION: Renal dysfunction was a significant and independent risk factor for death after endovascular aneurysm repair. PMID- 17117431 TI - Biowaiver monographs for immediate release solid oral dosage forms: isoniazid. AB - Literature data relevant to the decision to allow a waiver of in vivo bioequivalence (BE) testing for the approval of immediate release (IR) solid oral dosage forms containing isoniazid as the only active pharmaceutical ingredient (API) are reviewed. Isoniazid's solubility and permeability characteristics according to the Biopharmaceutics Classification System (BCS), as well as its therapeutic use and therapeutic index, its pharmacokinetic properties, data related to the possibility of excipient interactions and reported BE/bioavailability (BA) problems were taken into consideration. Isoniazid is "highly soluble" but data on its oral absorption and permeability are inconclusive, suggesting this API to be on the borderline of BCS Class I and III. For a number of excipients, an interaction with the permeability is extreme unlikely, but lactose and other deoxidizing saccharides can form condensation products with isoniazid, which may be less permeable than the free API. A biowaiver is recommended for IR solid oral drug products containing isoniazid as the sole API, provided that the test product meets the WHO requirements for "very rapidly dissolving" and contains only the excipients commonly used in isoniazid products, as listed in this article. Lactose and/or other deoxidizing saccharides containing formulations should be subjected to an in vivo BE study. PMID- 17117432 TI - Randomized clinical trial comparing feeding jejunostomy with nasoduodenal tube placement in patients undergoing oesophagectomy. AB - BACKGROUND: Feeding jejunostomy is frequently performed in patients undergoing oesophageal surgery, but can lead to serious complications. This prospective randomized trial compared the efficacy and complications of feeding jejunostomy with those of nasoduodenal tube feeding in oesophageal surgery. METHODS: Over an 18-month period, 150 consecutive patients undergoing oesophageal resection were randomized to participate in the trial. Enteral access was by jejunostomy in 79 patients and by nasoduodenal tube in 71. Enteral feeding was started on the first day after surgery. RESULTS: Full enteral feeding took 3 days to be established in both groups. Minor catheter-related complications occurred in 28 patients (35 per cent) in the jejunostomy group, and in 21 (30 per cent) in the nasoduodenal group (P = 0.488). One patient had jejunostomy leakage that required reoperation. Enteral nutrition was given for a median of 11 days in the jejunostomy group and for 10 days in the nasoduodenal group. Nine patients who had a jejunostomy and five with a nasoduodenal tube did not tolerate full enteral feeding (P = 0.411). CONCLUSION: Nasoduodenal tube feeding is safe and efficient after oesophageal resection. PMID- 17117433 TI - Screening for depression and anxiety disorders in primary care patients. AB - Mood and anxiety disorders are highly prevalent in primary health care. In this study we assessed performance of the Hospital Anxiety and Depression Scale (HADS) for screening of depression and anxiety disorders in a population of primary care patients. A total of 503 primary care patients consecutively admitted to the primary care medical center in Kaunas, Lithuania, completed the study. We found that the HADS subscale of depression (HADS-D) at a cutoff score of 6 or more showed the best performance screening for a major depressive episode diagnosed by means of the Mini International Neuropsychiatric Interview (MINI), with a sensitivity of 80%, specificity of 69%, positive predictive value of 80%, negative predictive value of 92%, and area under the receiver operating characteristic (ROC) curve of 0.75. Performance of the HADS-D against MINI diagnosis of dysthymia was weak. The HADS subscale of anxiety (HADS-A) at a cutoff score of 9 or more showed the best performance screening for MINI diagnosis of overall anxiety disorders, with a sensitivity of 77%, specificity of 75%, positive predictive value of 53%, negative predictive value of 90%, and area under the ROC curve of 0.76. These results suggest that in primary care patients HADS is an adequate screening instrument for the MINI diagnoses of major depressive episode, but not for dysthymia at a cutoff score of 6, and for anxiety disorders at a cutoff score of 9. PMID- 17117434 TI - Impact of polysol, a newly developed preservation solution, on cold storage of steatotic rat livers. AB - Chronic shortage of donor organs has led to acceptance of steatotic livers as grafts, although there is a higher risk of primary graft dysfunction. We herein report the beneficial impact of Polysol, a newly developed preservation solution, on cold storage of steatotic rat livers. Dietary hepatic steatosis was induced in Wistar rats by 2-day fasting and subsequent 3-day re-feeding with a fat-free, carbohydrate-rich diet. Fatty livers were retrieved, flushed and then stored at 4 degrees C for 24 hours with either HTK or Polysol. Functional integrity of the grafts was evaluated by isolated reperfusion with oxygenated Krebs-Henseleit buffer at 37 degrees C for 45 minutes in both groups. Polysol preservation resulted in significant reductions of not only parenchymal (AST (IU/L); 6728+/ 824 in HTK vs. 3107+/-718 in Polysol; P < 0.001) but also mitochondrial (GLDH (IU/L); 3189+/-773 vs. 1282+/-365; P < 0.01) enzyme release throughout reperfusion. Moreover, PVP (16.9+/-2.7 vs. 7.8+/-1.5 mmHg; P < 0.05), hepatic O2 consumption (0.291+/-0.047 vs. 1.056+/-0.053 micromol/g liver/min; P < 0.001), tissue ATP content (0.695+/-0.086 vs. 1.340+/-0.157 micromol/g dry-liver; P < 0.005), bile production (0.79+/-0.11 vs. 4.08+/-0.66 microL/g liver/45-min; P < 0.001), malondialdehyde into the perfusate (1.922+/-0.198 vs. 0.573+/-0.094 nmol/L; P < 0.0001) and wet/dry-weight ratio of the liver tissues (5.20+/-0.31 vs. 3.85+/-0.15; P < 0.005) were all better preserved by Polysol. In line with these benefits, electron microscopy revealed that Polysol preservation substantially suppressed deleterious mitochondrial alterations in steatotic livers. In conclusion, cold storage using Polysol resulted in significantly better integrity and function of steatotic livers. Polysol, therefore, may be a new alternative especially for "marginal" organs. PMID- 17117435 TI - Nitric oxide-dependence of satellite stem cell activation and quiescence on normal skeletal muscle fibers. AB - Satellite cells (quiescent precursors in normal adult skeletal muscle) are activated for growth and regeneration. Signaling by nitric oxide (NO) and hepatocyte growth factor (HGF) during activation has not been examined in a model that can distinguish quiescent from activated satellite cells. We tested the hypothesis that NO and HGF are required to regulate activation using the single fiber culture model. In normal fibers, HGF and inhibition of NO synthase (NOS) each increased activation without stretching, and NOS inhibition reduced stretch activation. Activation in unstretched mdx and NOS-I(-/-) fibers was three- to fourfold higher than normal, and was reduced by stretching. Distinctions were not due to different pax7-expressing populations on normal and mdx fibers. The population of c-met-expressing satellite cells on normal fibers was increased by stretch, demonstrating functional heterogeneity among normal satellite cells. Cycloheximide did not prevent the stretch-related increase in c-met expression, suggesting c-met may be an immediate-early gene in satellite cell activation. Results have important implications for designing new therapies that target the role of exercise in health, aging, and disease. PMID- 17117436 TI - Tbx1 regulation of myogenic differentiation in the limb and cranial mesoderm. AB - The T-box transcription factor Tbx1 has been implicated in DiGeorge syndrome, the most frequent syndrome due to a chromosomal deletion. Gene inactivation of Tbx1 in mice results in craniofacial and branchial arch defects, including myogenic defects in the first and second branchial arches. A T-box binding site has been identified in the Xenopus Myf5 promoter, and in other species, T-box genes have been implicated in myogenic fate. Here we analyze Tbx1 expression in the developing chick embryo relating its expression to the onset of myogenic differentiation and cellular fate within the craniofacial mesoderm. We show that Tbx1 is expressed before capsulin, the first known marker of branchial arch 1 and 2 muscles. We also show that, as in the mouse, Tbx1 is expressed in endothelial cells, another mesodermal derivative, and, therefore, Tbx1 alone cannot specify the myogenic lineage. In addition, Tbx1 expression was identified in both chick and mouse limb myogenic cells, initially being restricted to the dorsal muscle mass, but in contrast, to the head, here Tbx1 is expressed after the onset of myogenic commitment. Functional studies revealed that loss of Tbx1 function reduces the number of myocytes in the head and limb, whereas increasing Tbx1 activity has the converse effect. Finally, analysis of the Tbx1-mesoderm-specific knockout mouse demonstrated the cell autonomous requirement for Tbx1 during myocyte development in the cranial mesoderm. PMID- 17117437 TI - Loss of Fgfr3 leads to excess hair cell development in the mouse organ of Corti. AB - Previous studies have demonstrated the importance of FGF signaling at several stages in the development of the cochlea. At early stages of embryogenesis, Fgfr1, Fgfr2, and several FGFs are critical for both the induction of the otic vesicle and the initial development of the sensory epithelium. At late stages of cochlear development, Fgfr3 is necessary for the development of the tunnel of Corti. To determine the stage of development when Fgfr3 is required, we examined the expression of Fgfr3 and Fgf8 at various developmental stages. We also re examined the Fgfr3 -/- mouse with additional markers for developing supporting cells. We confirmed the previous analysis of the Fgfr3 -/- mice, indicating that there are deficiencies in support cell differentiation. Specifically, we find that the inner pillar cell never develops, while the outer pillar cell is stalled in its differentiation. In addition, we found an extra row of outer hair cells, and accompanying Deiters' cells, in the apical two thirds of the organ of Corti in the Fgfr3 mutant. Thus, in addition to controlling the fate decision between pillar cells and Deiters' cells, we find that Fgfr3 also regulates the width of the sensory epithelium. PMID- 17117438 TI - Inhibition of Sonic hedgehog signaling leads to posterior digit loss in Ambystoma mexicanum: parallels to natural digit reduction in urodeles. AB - Molecular mechanisms patterning the tetrapod limb, including anterior-posterior axis determination involving Sonic hedgehog (Shh), have received much attention, particularly in amniotes. Anterior-posterior patterning in urodele amphibians differs radically from that of amniotes in that it shows a pronounced anterior-to posterior sequence of digit development. In contrast, amniotes develop their digits almost simultaneously with a slight posterior-to-anterior polarity. Here we use cyclopamine, an inhibitor of the Hedgehog signaling pathway, to investigate the role of Shh in anterior-posterior patterning in the urodele limb. Inhibition of Shh signal transduction affects digit number long before their morphological appearance. In accordance with the apparently derived order of digit development in urodeles, exposure reproducibly removes digits in a posterior-to-anterior sequence, the inverse of their developmental sequence. This pattern of digit loss mimics the order of digit loss in natural variation. We suggest that variation in Shh expression and/or signal transmission may explain natural variation in digit number in urodeles. PMID- 17117439 TI - BMP type I receptor ALK2 is essential for proper patterning at late gastrulation during mouse embryogenesis. AB - Bone morphogenetic proteins (BMPs) have multiple functions during vertebrate development. Previously, it was shown that BMP type I receptor ALK2 (also known as ACVRI, ActRI, or ActRIA) was important for normal mouse gastrulation by deleting exon 4 or exon 5 of Alk2. Recently, flanking exon 7 by loxP sites generated a conditional allele for Alk2. To assess whether the deletion of exon 7 causes functional null of ALK2, and does not produce a dominant negative form or a partially functional form of ALK2, we performed a comparative analysis between Alk2 homozygous mutant embryos with an exon 5 deletion (Alk2(Delta5/Delta5)) and embryos with an exon 7 deletion (Alk2(Delta7/Delta7)). Both Alk2(Delta5/Delta5) and Alk2(Delta7/Delta7) mutants showed identical morphological gastrulation defects. Histological examinations and molecular marker analyses revealed identical abnormal gastrulation phenotypes in Alk2(Delta5/Delta5) and Alk2(Delta7/Delta7) mutants. Although Fgf8 was expressed in the primitive streak of Alk2(Delta5/Delta5) and Alk2(Delta7/Delta7) mutants, Brachyury, Wnt3a, and Tbx6 were dramatically downregulated in Alk2(Delta5/Delta5) and Alk2(Delta7/Delta7) mutants. These results indicate that deletion of exon 7 for Alk2 leads to a functionally null mutation in vivo, and Alk2 is crucial for sustaining the proper gastrulation events in early mouse embryogenesis. PMID- 17117441 TI - Prox1 expression patterns in the developing and adult murine brain. AB - Prox1, a homeobox gene related to the Drosophila gene prospero, is necessary for retina, lens, liver, pancreas, and lymphatics development. However, not much is yet known about Prox1 expression during central nervous system development. Here we provide a detailed analysis of Prox1 mRNA and protein expression during prenatal and postnatal murine brain development. Prenatally, Prox1 is expressed in the subventricular zone or in early differentiating regions of the brain. At these stages, Prox1 mRNA, but not Prox1 protein, was also detected in several regions of the prethalamus and hypothalamus. At an early postnatal stage, Prox1 expression is mainly detected in several nuclei of the thalamus, the cerebellum, and the hippocampus. In adulthood, Prox1 expression remains only in the hippocampus and cerebellum. These complex patterns of expression suggest that Prox1 activity is differentially required during brain development and adulthood. PMID- 17117442 TI - Quantification of trandolapril and its metabolite trandolaprilat in human plasma by liquid chromatography/tandem mass spectrometry using solid-phase extraction. AB - A sensitive high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (MS/MS) method was developed and validated for the simultaneous quantification of trandolapril and its metabolite trandolaprilat in human plasma using ramipril as an internal standard. Following solid-phase extraction, the analytes were separated using an isocratic mobile phase on a reversed-phase column and analyzed by MS/MS in the multiple reaction monitoring mode using the respective [M-H]- ions, m/z 429/168 for trandolapril, m/z 401/168 for trandolaprilat and m/z 415/166 for the internal standard. The method exhibited a linear dynamic range of 20-10,000 pg/mL for both trandolapril and trandolaprilat in human plasma. The lower limit of quantification was 20 pg/mL for both trandolapril and its metabolite. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. A run time of 2.0 min for each sample made it possible to analyze more than 400 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic, bioavailability or bioequivalence studies. PMID- 17117443 TI - Atmospheric pressure desorption/ionization on silicon ion trap mass spectrometry applied to the quantitation of midazolam in rat plasma and determination of midazolam 1'-hydroxylation kinetics in human liver microsomes. AB - The application of atmospheric pressure desorption/ionization on silicon (AP DIOS) coupled with ion trap mass spectrometry (ITMS) was investigated for the quantification of midazolam in rat plasma, and determination of midazolam 1' hydroxylation kinetics in pooled human liver microsomes. Results indicate good sensitivity with absolute detection limits for midazolam in rat plasma of approximately 300 femtograms. A linear dynamic range from approximately 10-5000 ng/mL was obtained in rat plasma with analysis times of 1 min per sample. Kinetic constants for midazolam 1'-hydroxylation in human liver microsomes yielded an apparent Km of 10.0 microM and Vmax of 6.4 nmol/min/mg. Studies investigating the inhibition of 1'-hydroxymidazolam formation by the cytochrome P450 3A4 model inhibitor ketoconazole yielded an IC50 of 0.03 microM. Quantitative precision for replicate analysis of rat plasma and human liver microsomal samples was variable with relative standard deviation (RSD) values ranging from a low of approximately 3% to over 50%, with the highest variability observed in data from human liver microsomal incubations. While preliminary studies investigating the application of AP-DIOS-ITMS suggested feasibility of this technique to typical pharmacokinetic applications, further work is required to understand the underlying causes for the high variability observed in these investigations. PMID- 17117444 TI - Patient-reported outcome I: measuring what matters in musculoskeletal care. AB - Patients have an important role to play in communicating the impact of disease and the effectiveness of health care. Well-developed patient-reported outcome measures (PROMs) can provide a clinically relevant and scientifically rigorous resource for including the patient perspective in decisions about health care and subsequent evaluation. This article will introduce readers to the field of health outcome assessment, and specifically to patient-reported outcome measures (PROMs). The reasons for measuring health, different types of PROMs and potential applications will be discussed. A second article will present important information for users of PROMs to consider when selecting and using PROMs in research, routine practice or quality assurance. PMID- 17117445 TI - The impact of anti-tumour necrosis factor therapy for rheumatoid arthritis on the use of other drugs and hospital resources in a pragmatic setting. AB - BACKGROUND: Anti-tumour necrosis factor (anti-TNF) therapy has been an important development for the treatment of rheumatoid arthritis (RA) but the impact of its delivery on hospital resources in still emerging. AIMS: We audited the effect of starting anti-TNF on the use of other anti-rheumatic therapies and hospital resources in a routine secondary care setting. METHODS: A retrospective study of resource use before and after anti-TNF was conducted. Hospital records of 54 RA patients were studied and data taken from the time of commencing anti-TNF to 1 October 2004 and an equal time period prior to commencing anti-TNF. Identical data were collected for 54 controls not on anti-TNF. Relevant figures were extrapolated to per annum rates. Results were analysed using two-factor ANOVAs comparing the pre- versus post-anti-TNF period. Cases on intravenous (IV) versus subcutaneous (SC) anti-TNF were also compared in separate ANOVAs. RESULTS: Mean duration of anti-TNF therapy was 17.04 months (range 3.60-42.36). Mean pre- and 3 months post-anti-TNF Disease Activity Scores (DAS28) were 6.93 and 3.88, respectively. Cases were more likely than controls to be on oral prednisolone pre and post-anti-TNF. Methylprednisolone requirement, number of disease-modifying anti-rheumatic drugs (DMARDs), telephone helpline contacts and duration as an inpatient reduced significantly post-anti-TNF. Day case admissions increased but outpatient appointments decreased only in cases on IV anti-TNF. CONCLUSIONS: In a pragmatic setting, anti-TNF therapy led to reduced need for steroid injections and other DMARDs, as well as reductions in use of several hospital resources. Wider replication of these findings will be important for planning delivery. PMID- 17117446 TI - Influences that drive clinical decision making among junior rheumatology nurses: a qualitative study. AB - This paper presents a qualitative study exploring the influences that drive clinical decision making among a small group of junior rheumatology nurses.A qualitative, descriptive design was chosen. Semi-structured interviews were used with a purposive sample of six junior staff nurses from two inpatient rheumatology wards in a large teaching hospital in the North of England. The interviews were audiotaped and transcribed using Burnard's (1991) thematic content analysis. The findings demonstrate four distinct themes which influence clinical decision making including professional development, patient-focused care, working in a specialty and rheumatology nursing. Development of experiential knowledge alongside access to specialized information and expert practitioners was also influential in informing decisions. PMID- 17117447 TI - Evidence-based review of shoulder pain. PMID- 17117448 TI - A benzimidazole-based chromogenic anion receptor. AB - A benzimidazole-based anion receptor 1 has been designed and synthesized. The structure of 1 is confirmed through single crystal X-ray diffraction analysis. Anion-binding studies carried out using (1)H NMR and UV-visible revealed that this compound exhibits selective recognition toward F(-) over other halide anions. The highest selectivity for F(-) among the halides is attributed mainly to the strongest hydrogen-bond interaction of the receptor with F(-); in addition, the higher match in geometry between the receptor and F(-) also plays a role in the selective recognition of the receptor for F(-). Receptor 1 (1 x 10( 5) M) shows dramatic color change from light-yellow to red in the presence of tetrabutylammonium fluoride (TBAF) (1 x 10(-5) M). Moreover, F(-)-induced color changes remain the same even in the presence of a large excess of Cl(-), Br(-), I(-). PMID- 17117449 TI - Immunostimulating effects of the polyphenol-rich fraction of sugar cane (Saccharum officinarum L.) extract in chickens. AB - The phagocytic activity of peripheral blood leukocytes (PBL) in chickens orally administered sugar cane extracts (SCE) or polyphenol-rich fraction (PRF) of SCE (500 mg/kg/day) for 3 consecutive days increased significantly, when compared with that of saline-administered control chickens. Chickens orally administered SCE or PRF (500 mg/kg/day) for 3 consecutive days showed significantly higher antibody responses against sheep red blood cells and Brucella abortus than control chickens. In addition, oral administration of SCE or PRF also resulted in a significant increase in the number of IgM- and IgG-plaque forming cell responses of PBL, intestinal leukocytes and splenocytes, when compared with those of control chickens. Furthermore, delayed type hypersensitivity responses to human gamma globulin significantly increased in chickens orally administered SCE or PRF, compared with those of control chickens when evaluated on the basis of net increased wattle thickness at 24, 48 and 72 h after challenge. These results suggest that PRF of SCE has an immunostimulating effect in chickens. PMID- 17117450 TI - Doublet-quartet conversion in negative ions as a possible mechanism of the electron autodetachment delay. AB - Molecular negative ions with abnormally high lifetimes (20 micros) relative to electron autodetachment were registered at 1.65 eV electron energy in pyromellitic acid imide by means of negative ion mass spectrometry with resonant electron capture. Using quantum-chemical calculations, it was shown that the delay of the electron autodetachment may be caused by the conversion of an originally generated molecular ion-doublet into a molecular ion-quartet, as a result of intersystem crossing of the potential energy surfaces of these ions. It was noticed that the ion-quartet cannot decay rapidly into the molecule of the ground state because of the prohibition of the spin flip, which is similar to that for phosphorescence. It can also not decay into the parent triplets, because these triplets are higher in energy than the ion. As a whole, both prohibitions provide the observed effect of the high lifetime of the ions. PMID- 17117451 TI - Kamikihi-to, a Kampo medicine, ameliorates impairment of spatial memory in rats. AB - The present study investigated the effects of Kamikihi-to (KKT), a Kampo medicine, on impairment of spatial memory in rats using an eight-arm radial maze task. Scopolamine (0.5 mg/kg, i.p.), a non-selective muscarinic receptor antagonist, and Delta(9)-tetrahydrocannabinol (THC; 6 mg/kg, i.p.), a principal psychoactive component of marihuana, each markedly impaired the spatial memory. KKT (1 and 3 mg/kg, p.o.) significantly improved the scopolamine-induced impairment of spatial memory. KKT (30 mg/kg, p.o.) also improved significantly the THC-induced impairment of spatial memory. Moreover, KKT (3 and 30 mg/kg, p.o.) enhanced tremors induced by oxotremorine, a muscarinic M(1) receptor agonist. Taken together these findings suggest that KKT is a useful drug for treating memory deficits. PMID- 17117452 TI - Intellectual property protection in the natural product drug discovery, traditional herbal medicine and herbal medicinal products. AB - Traditional medicine is an important part of human health care in many developing countries and also in developed countries, increasing their commercial value. Although the use of medicinal plants in therapy has been known for centuries in all parts of the world, the demand for herbal medicines has grown dramatically in recent years. The world market for such medicines has reached US $ 60 billion, with annual growth rates of between 5% and 15%. Researchers or companies may also claim intellectual property rights over biological resources and/or traditional knowledge, after slightly modifying them. The fast growth of patent applications related to herbal medicine shows this trend clearly. This review presents the patent applications in the field of natural products, traditional herbal medicine and herbal medicinal products. Medicinal plants and related plant products are important targets of patent claims since they have become of great interest to the international drug and cosmetic industry. PMID- 17117453 TI - Effects of phytoestrogenic isoflavones from red clover (Trifolium pratense L.) on experimental osteoporosis. AB - The most common type of osteoporosis is bone loss associated with ovarian hormone deficiency at menopause. There is evidence that diets which contain high levels of phytoestrogenic isoflavones are associated with a low incidence of osteoporosis and menopausal symptoms. Plant extracts, which contain high levels of isoflavones, such as Red clover (Trifolium pratense L.), have been used to reduce menopausal symptoms. The objective of this study was to evaluate the preventive effects of Red clover total isoflavones on the progression of bone loss induced by estrogens deficiency (ovariectomy) in rats. Bilateral ovariectomy was performed on female Wistar rats. One week after the operation the rats were treated with an oral dose of 20 and 40 mg of total isoflavones daily for 14 weeks. The results from this study showed that the ovariectomy reduced bone mineral content, femoral weight, femoral density, mechanical strength of the tibia and increased the levels of bone specific alkaline phosphatase in the serum and the number of osteoclasts in the femur sections compared with sham operated controls. Treatment with isoflavones significantly increased bone mineral content, mechanical strength of the tibia, femoral weight, femoral density and prevented the rise of serum alkaline phosphatase levels. In addition, the treatment with isoflavones significantly reduced the number of osteoclasts compared with the ovariectomized control rats. These findings suggest that Red clover isoflavones are effective in reducing bone loss induced by ovariectomy, probably by reducing of the bone turnover via inhibition of bone resorption. PMID- 17117454 TI - In vitro metabolism of ciclesonide in human nasal epithelial cells. AB - Ciclesonide, a corticosteroid in development for allergic rhinitis, is converted to the pharmacologically active metabolite, desisobutyryl-ciclesonide (des-CIC), and des-CIC is subsequently esterified with fatty acids. Various experiments were performed to investigate ciclesonide metabolism in human nasal epithelial cells (HNEC). Human nasal epithelial cells were incubated with (a) 0.1 microM ciclesonide for 1 h and medium without ciclesonide for up to 24 h, (b) esterase inhibitors for 0.5 h followed by 5 microM ciclesonide for 6 h, or (c) 1 microM des-CIC for 6 h followed by medium without des-CIC for up to 24 h. Ciclesonide, des-CIC and des-CIC fatty acid conjugate concentrations were determined by high performance liquid chromatography with tandem mass spectrometry. The amount of ciclesonide in HNEC decreased approximately 93-fold from 0.5 to 24 h. In contrast, des-CIC was present at constant levels throughout the post-treatment period. Furthermore, fatty acid conjugates of des-CIC were retained in HNEC up to 24 h post-treatment. Carboxylesterase and cholinesterase inhibitors decreased ciclesonide metabolism > or =50%. The total amounts of des-CIC fatty acid conjugates decreased and the extracellular amounts of des-CIC increased with time. In conclusion, ciclesonide was rapidly converted to des-CIC by carboxylesterases and cholinesterases, and des-CIC underwent reversible fatty acid conjugation in HNEC. PMID- 17117455 TI - Assessment of dose proportionality of muraglitazar after repeated oral dosing in rats via a sparse sampling methodology. AB - Muraglitazar is an alpha/gamma-dual peroxisome proliferator-activated receptor (PPAR) agonist. This study evaluated the single- and multiple-dose oral toxicokinetics of muraglitazar in rats at doses of 3, 30 and 300 mg/kg/day. In total, 15 rats/gender/dose group received muraglitazar every day for 1 month. On both day 1 and day 28, blood samples were obtained at 0.5, 2, 4, 6, 8 and 24 h post-dose, followed by LC/MS analysis. In order to minimize blood loss in the rats, a sparse sampling approach was used to delineate the toxicokinetics. The peak plasma concentration (C(max)) and area under the plasma concentration-time curve (TAUC(0-t)) values for muraglitazar increased in a proportion less than the increment in dose. As the dose increased in the ratio 1:10:100, the C(max) for muraglitazar in male and female rats increased in the ratio of 1:10.3:58.6 and 1:15.3:75.3 on day 1, and 1:5.9:28.1 and 1:13.3:37.3 on day 28, respectively. The corresponding TAUC(0-t) values for males and females were in the ratio of 1:10.2:131 and 1:12.4:131 on day 1, and 1:9.5:93.4 and 1:11.8:94.3 on day 28, respectively. The results indicate that muraglitazar exhibits a dose dependent toxicokinetics in rats and the systemic exposure of muraglitazar was decreased on day 28 compared with day 1. PMID- 17117456 TI - Patterning microbeads inside poly(dimethylsiloxane) microfluidic channels and its application for immobilized microfluidic enzyme reactors. AB - We propose a convenient and reliable approach for immobilizing microbeads on poly(dimethylsiloxane) (PDMS) microchips. It is built upon a simple fabrication procedure of PDMS chip through directly printing the master with an office laser printer which was described in our previous work (J. Chromatogr. A 2005, 1089, 270-275). On the printed toners used as the positive relief of the master, microbeads were immobilized by a thermal treatment and then transferred to the surface of the microchip by direct molding of the prepolymer on the master. With this approach, the region-selective immobilization of microbeads and the fabrication of PDMS microchips can be accomplished at the same time. Then, using these microbeads as supports, further modification with enzyme was achieved. Surface characteristics of the microbeads-modified PDMS microchannels were investigated with scanning electron microscope, atomic force microscope, and inverse fluorescence microscope. The electrokinetic properties of the native PDMS and the modified PDMS chips were also compared. Based on this approach, an immobilized glucose oxidase (GOD) reactor was constructed and the reaction using glucose as substrate was studied. All these experiments aim to show that the proposed approach may have a good potential in the study of biochemistry and other related areas. PMID- 17117457 TI - Integration of nanoporous membranes for sample filtration/preconcentration in microchip electrophoresis. AB - Microfluidic devices integrating membrane-based sample preparation with electrophoretic separation are demonstrated. These multilayer devices consist of 10 nm pore diameter membranes sandwiched between two layers of PDMS substrates with embedded microchannels. Because of the membrane isolation, material exchange between two fluidic layers can be precisely controlled by applied voltages. More importantly, since only small molecules can pass through the nanopores, the integrated membrane can serve as a filter or a concentrator prior to microchip electrophoresis under different design and operation modes. As a filter, they can be used for separation and selective injection of small analytes from sample matrix. This has been effectively applied in rapid determination of reduced glutathione in human plasma and red blood cells without any off-chip deproteinization procedure. Alternatively, in the concentrator mode, they can be used for online purification and preconcentration of macromolecules, which was illustrated by removing primers and preconcentrating the product DNA from a PCR product mixture. PMID- 17117458 TI - Quantitative analysis of amyloid beta peptides in cerebrospinal fluid of Alzheimer's disease patients by immunoaffinity purification and stable isotope dilution liquid chromatography/negative electrospray ionization tandem mass spectrometry. AB - The 40 and 42 amino-acid residue forms of amyloid beta (Abeta(1-40) and Abeta(1 42)) in cerebrospinal fluid (CSF) have been proposed as potential biomarkers of Alzheimer's disease (AD). Quantitative analyses of Abeta peptides in CSF have relied almost exclusively on the use of immunoassay-based assays such as the enzyme-linked immunosorbent assay (ELISA) procedure. However, due to the ability of the Abeta peptides to readily self-aggregate or bind to other proteins and glassware, such analyses are extremely challenging. Analyses are further complicated by the potential of the peptides to undergo post-translational modifications and the possibilities for cross-reaction in the ELISA assays with endogenous components of the CSF. An approach based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) has now been developed which overcomes these methodological issues. The key steps in implementing this new approach involved immunoaffinity purification coupled with the use of [15N] labeled Abeta peptides as internal standards, a basic LC mobile phase, negative ion electrospray ionization, and a basic solvent for dissolving the peptides and washing the injection needle to prevent carryover of analytes during multiple injections on the LC/MS system. The validated method had limits of quantitation of 44 fmol/mL (200 pg/mL) for Abeta(1-42) and 92 fmol/mL (400 pg/mL) for Abeta(1 40). An excellent correlation was found between the LC/MS/MS assay and an ELISA assay for Abeta(1-42) in human CSF (r2 = 0.915), although less correlation was observed for Abeta(1-40) (r2 = 0.644). Mean CSF Abeta(1-42) concentrations for samples collected 2 weeks apart from a limited number of AD patients provided additional confidence in the reproducibility of the LC/MS/MS assay. Concentrations for duplicate samples from AD patients were slightly higher than most previously reported values (mean 1.06 +/- 0.25 ng/mL; n = 7). Abeta(1-40) concentrations in duplicate samples obtained from AD patients were also reproducible but were found to be slightly lower than most previously reported values (mean 6.36 +/- 3.07 ng/mL; n = 7). Consistent with literature reports, mean Abeta(1-42) concentrations were found to be lower in AD patients compared with the normal subjects (mean 1.49 +/- 0.59 ng/mL; n = 7), whereas there was no difference in Abeta(1-40) concentrations between AD patients and normal subjects (mean 5.88 +/- 3.03 ng/mL; n = 7). The accuracy and precision of the LC/MS assay mean that it will be a useful complement to existing ELISA assays for monitoring therapeutic interventions designed to modulate CSF Abeta(1-42) concentrations in individual AD patients. Moreover, the introduction of stable isotope labeled internal standards offers the potential to achieve a more rigorous account of the influence of methodological effects related to sample collection and processing. PMID- 17117459 TI - Sorting and pooling strategy: a novel tool to map a virus proteome for CD8 T-cell epitopes. AB - Human cytomegalovirus (CMV) is a major cause of morbidity in immunocompromised individuals. However, no efficient vaccine has been developed to date. Identification of T-cell target proteins and epitopes is crucial not only for developing a successful immunization strategy, but also for new approaches using adoptive transfer of antigen-specific T-cells. The CMV genome has more than 200 open reading frames potentially coding for as many proteins. Here, we describe a robust, fast, and simple SPOT synthesis strategy, which allowed us to micro synthesize every possible CD8 T-cell epitope in the entire potential CMV proteome. So far, 9069 of these peptides have been tested in an ex vivo T-cell stimulation assay. As well as confirming a number of previously known epitopes, we identified several new ones. PMID- 17117460 TI - Beta-amino acid analogs of an insect neuropeptide feature potent bioactivity and resistance to peptidase hydrolysis. AB - Insect neuropeptides of the insect kinin class share a common C-terminal pentapeptide sequence F(1)X(1)(2)X(2)(3)W(4)G(5)-NH(2) (X(2)(3) = P, S, A) and regulate such critical physiological processes as water balance and digestive enzyme release. Analogs of the insect kinin class, in which the critical residues of F(1), P(3), and W(4) were replaced with beta(3)-amino acid or their beta(2) homo-amino acid variants, have been synthesized by the solid phase peptide strategy. The resulting single- and double-replacement analogs were evaluated in an insect diuretic assay and enzyme digestion trials. Analogs modified in the core P(3) position produce a potent and efficacious diuretic response that is not significantly different from that obtained with the endogenous achetakinin peptides. The analogs also demonstrate enhanced resistance to hydrolysis by ACE and NEP, endopeptidases that inactivate the natural insect neuropeptides. This paper describes the first instance of beta-amino acids analogs of an arthropod peptide that demonstrate significant bioactivity and resistance to peptidase degradation. PMID- 17117461 TI - Simultaneous determination of natural and synthetic estrogens by EKC using a novel microemulsion. AB - A novel microemulsion based on sodium bis(2-ethylhexyl) sulfosuccinate (AOT) was developed for the simultaneous determination of natural and synthetic estrogens by microemulsion EKC (MEEKC). The microemulsion system consisted of 1.4% w/w AOT, 1.0% w/w octane, 7.0% w/w 1-butanol and 90.6% w/w 20 mM sodium salt of 3 (cyclohexylamino)-2-hydroxy-1-propanesulfonic acid (CAPSO) and 10 mM phosphate buffer at pH 12.5. A baseline resolution in the separation of estrone, 17beta estradiol, estriol, estradiol 17-hemisuccinate, etinilestradiol, estradiol 3 benzoate, and estradiol 17-valerate was achieved in comparison to the traditional MEEKC system with SDS in less than 15 min. The optimized electrophoretic conditions included the use of an uncoated-silica capillary of 60 cm of total length and 75 microm id, an applied voltage of 25 kV, a temperature of 25 degrees C and 214 UV-detection. Parameters of validation such as specificity, linearity, accuracy, LOD, LOQ and robustness were evaluated according to international guidelines. Due to its simplicity, accuracy, and reliability, the proposed method can be an advantageous alternative to the traditional methodologies for the analysis of natural and synthetic estrogens in different pharmaceutical forms. PMID- 17117462 TI - CE-LIF analysis of mitochondria using uncoated and dynamically coated capillaries. AB - This report is the first demonstration of the use of uncoated and dynamically coated capillaries for the separation of individual mitochondria via CE. Currently, the analysis of individual mitochondria relies upon fused-silica capillaries coated with a hydrophilic polymer (e.g. poly(acryloylaminopropanol)), which is used to minimize adsorption to the capillary surface. Both uncoated fused-silica capillaries and 0.2% w/w poly(vinyl alcohol) dynamic coating solutions are used to successfully analyze isolated individual mitochondrial particles using CE-LIF. While it was possible to separate mouse liver mitochondria on an uncoated capillary, rat liver mitochondria proved to have strong adsorption characteristics that only allowed them to be adequately separated with a PVA dynamic coating or a poly(acryloylaminopropanol) (AAP) capillary. The possible causes for this adsorption are analyzed and discussed. This study shows that uncoated and dynamically coated capillaries can be used in place of AAP-coated capillaries to analyze mitochondria and suggests the use of these capillaries for the analysis of other organelles, offering a greatly simplified method for the analysis of individual organelles. PMID- 17117465 TI - Single-molecule measurements of trapped and migrating circular DNA during electrophoresis in agarose gels. AB - The effect of agarose gel concentration and field strength on the electrophoretic trapping of open (relaxed) circular DNA was investigated using microscopic measurements of individual molecules stained with a fluorescent dye. Three open circles with sizes of 52.5, 115, and 220 kbp were trapped by the electric field (6 V/cm) and found to be predominately fixed and stretched at a single point in the gel. The length of the stretched circles did not significantly change with agarose concentration of the gels (mass fractions of 0.0025, 0.01, and 0.02). The relaxation kinetics of the trapped circles was also measured in the gels. The relaxation of the large open circles was found to be a slow process, taking several seconds. The velocity and average length of the 52.5 kbp open circles and 48.5 kbp linear DNA were measured during electrophoresis in the agarose gels. The velocity increased when the agarose concentrations were lowered, but the average length of the open-circle DNA (during electrophoresis) did not significantly change with agarose gel concentrations. The circles move through the gels by cycles of stretching and relaxation during electrophoresis. Linear dichroism was also used to investigate the trapping and alignment of the 52.5 kbp open circles. The results in this study provide information that can be used to improve electrophoretic separations of circular DNA, an important form of genetic material and commonly used to clone DNA. PMID- 17117466 TI - Stability of and inflammatory response to silicon coated with a fluoroalkyl self assembled monolayer in the central nervous system. AB - A self-assembled monolayer (SAM) of fluoroalkyl silane (FAS) (CF(3)(CF(2))(5)(CH(2))(2)SiCl(3)) was deposited on the surface of silicon wafers, aiming to enhance its stability and to reduce the inflammatory response in the central nervous system. Following implantation of the FAS SAM coated silicon in rat brains, the FAS SAM coating failed to reduce the inflammatory response of silicon, because it could not prevent the corrosion of the underlying silicon. The corrosion was hindered for the initial 10 days by the FAS SAM coating, but there was nearly no difference when compared to the uncoated silicon when the implantation periods were extended to 28 and 90 days. The FAS SAM coating was completely removed within 28 and 90 days. Meanwhile, on all the extracted uncoated and FAS SAM coated silicon wafers, there were proteinaceous substances deposited on the surfaces, and the amount of the deposits increased with exposure time. PMID- 17117467 TI - Biodegradation of high-toughness double network hydrogels as potential materials for artificial cartilage. AB - This study evaluated biodegradation properties of four novel high-toughness double network (DN) hydrogels as potential materials for artificial cartilage. Concerning each DN gel material, a total of 12 specimens were prepared, and 6 of the 12 specimens were examined to determine the mechanical properties without any treatments. The remaining 6 specimens were implanted into the subcutaneous tissue, using 6 mature female rabbits. At 6 weeks after implantation, the mechanical properties and the water content of the implanted specimens were measured. In the poly(2-acrylamide-2-methyl-propane sulfonic acid)/poly(N,N' dimethyl acrylamide) DN gel, the ultimate stress and the tangent modulus were significantly increased from 3.10 and 0.20 MPa, respectively, to 5.40 and 0.37 MPa, respectively, with a significant reduction of the water content after implantation (94 to 91%). In the poly(2-acrylamide-2-methyl-propane sulfonic acid)/polyacrylamide DN gel and the cellulose/poly(dimethyl acrylamide) DN gel, the stress (11.4 and 1.90 MPa, respectively) and the modulus (0.30 and 1.70 MPa, respectively) or the water content rarely changed after implantation (90 and 85%, respectively). In the bacterial cellulose/gelatin DN gel, the ultimate stress was dramatically reduced from 4.30 to 1.98 MPa with a significant increase of the water content after implantation (78 to 86%). This study implied that these DN gels except for the cellulose/gelatin DN gel are potential materials that may meet the requirements of artificial cartilage. PMID- 17117468 TI - Biomedical coatings based on chitin soluble extract for inhibition of fungal adhesion to polymeric surfaces. AB - Indwelling medical devices made of polymeric materials, such as intravenous (IV) catheters, are known risk factors for development of fungal infection, particularly systemic candidiasis, that are a significant cause of morbidity and lethality in compromised patients. Candida can form a biofilm on the polymeric surface, serving as a nidus for systemic, difficult to eradicate infection. The current research focuses on development and study of a chitin soluble extract (CSE) coating on polyurethane (PU), in order to reduce the level of adherence of C. albicans to the PU surface. The immobilization of CSE onto the PU surface was performed using both, chemical binding and physical adsorption. Our results indicate that CSE develops a unique tertiary structure in which basic elements in the range of 100 nm build "fingers" and these "fingers" are arranged in a concentric structure around a center. The CSE coated films showed 75% inhibition of C. albicans adhesion for the chemical binding and 83% for the physical adsorption coating and even after 11 weeks the inhibitory effect on adhesion is still significant. Hence, our new coatings may lead to a new generation of medical devices with surfaces that can prevent fungal adhesion. PMID- 17117469 TI - Controlled release of nerve growth factor from fibrin gel. AB - Nerve growth factor (NGF) is known to promote the axonal regeneration in injured nerve system. Delivery of NGF for a long period in a controlled manner may enhance the regeneration efficacy. In this study, we investigated whether NGF can be released from fibrin gel for a long period in a controlled manner. We also investigated whether sustained delivery of NGF using fibrin gel can enhance the efficacy of NGF in vitro. The addition of heparin to fibrin gel decreased the rate of NGF release from the fibrin gel. As the concentrations of thrombin and fibrinogen in fibrin gel increased, the NGF release rate decreased significantly, and the initial release burst decreased. NGF was released for up to 14 days in vitro. The bioactivity of NGF released from fibrin gel was assessed by morphological changes of pheochromocytoma (PC12) cells cultured in the presence of NGF-containing fibrin gel. NGF released from fibrin gel exhibited significantly higher degrees of PC12 cell viability and differentiation than NGF added in a free form daily into the culture medium. This study demonstrates that fibrin gel can release NGF in a sustained, controlled manner and in a bioactive form. PMID- 17117470 TI - Hybrid composites of calcium phosphate granules, fibrin glue, and bone marrow for skeletal repair. AB - Synthetic bone substitutes, such as calcium phosphate ceramics, give good results in clinical applications. In order to adapt to surgical sites, bioceramics come in the form of blocks or granules, and are either dense or porous. Combining these bioceramics with fibrin glue provides a mouldable and self-hardening composite biomaterial with the biochemical properties of each component. Critical sized defects in the femoral condyle of rabbits were filled with TricOs/fibrin glue/bone marrow hybrid/composite material. The TricOs granules (1-2 mm) were composed of hydroxyapatite and beta tricalcium phosphate (60/40 in weight). The fibrin glue was composed of fibrinogen, thrombin and other biological factors and mixed with MBCP granules either simultaneously or sequentially. Bone marrow was also added to the MBCP/fibrin composite prior to filling the defects. After 3, 6, 12, and 24 weeks of implantation, the newly-formed bone was analysed with histology, histomorphometry and mechanical tests. The newly-formed bone had grown centripetally. Simultaneous application of fibrin glue showed better results for mechanical properties than sequential application after 6 weeks. Around 40% of bone had formed after 24 weeks in the three groups. Although the addition of bone marrow did not improve bone formation, the MBCP/fibrin material could be used in clinical bone filling applications. PMID- 17117471 TI - Fabrication and characterization of a novel composite PNIPAAm hydrogel for controlled drug release. AB - Novel composite poly(N-isopropylacrylamide) (PNIPAAm) hydrogels, containing poly(N-isopropylacrylamide)-block-poly(methyl methacrylate) (PNIPAAm-b-PMMA) micelles for sustained drug delivery were prepared and characterized. Various amounts of thermal sensitive PNIPAAm-b-PMMA micelles were incorporated physically into thermosensitive PNIPAAm bulk hydrogel to form composite PNIPAAm hydrogels. The resultant composite hydrogels were characterized by scanning electron microscopy (SEM) and differential scanning calorimetry (DSC) for morphological and thermal properties, respectively. The temperature dependence of swelling ratio and response kinetics upon heating or cooling were also investigated to understand the smart properties, that is, temperature-sensitive properties of the composite PNIPAAm hydrogels. These composite PNIPAAm hydrogels exhibited a faster shrinking kinetics than the one of pure PNIPAAm hydrogels. In addition, prednisone acetate, used as model drug, was loaded into the micelles incorporated to the composite PNIPAAm hydrogels. The controlled release behavior of the novel composite PNIPAAm hydrogels at different temperatures (22 and 37 degrees C) was examined. PMID- 17117472 TI - Paclitaxel-loaded composite fibers: microstructure and emulsion stability. AB - New core/shell fiber structures loaded with paclitaxel were developed and studied. These composite fibers are ideal for forming thin, delicate, biomedically important structures for various applications. Possible applications include fiber-based endovascular stents that mechanically support blood vessels while delivering drugs for preventing restenosis directly to the blood vessel wall, or drug delivery systems for cancer treatment. The core/shell fiber structures were formed by "coating" nylon fibers with porous paclitaxel containing poly(DL-lactic-co-glycolic acid) structures. Shell preparation ("coating") was performed by freeze-drying water in oil emulsions. The present study focused on the effects of the emulsion's formulation (composition) and processing conditions on the porous shell structure, which actually reflects the emulsion's stability and also the drug release profile from the fibers. In general, extremely porous "shell" structures were obtained with good adhesion to the core fiber. An increase in the emulsion's drug content and copolymer composition demonstrated a significant effect on pore size and distribution, because of enhanced emulsion instability, whereas the homogenization rate and duration had only a slight effect on the pores' microstructure. The thermodynamic parameters in the studied system are thus more important than the kinetic parameters in determining the emulsion's stability and the shell's porous structure. PMID- 17117473 TI - A novel methodology for imaging new bone formation around bioceramic bone substitutes. AB - In this study, Frost's bulk-staining in combination with confocal laser scanning microscopy (CLSM) was used to image and characterize ground sections of undecalcified rat bone tissue with in situ bioceramic implants. This was addressed by bulk staining specimens in alcohol-soluble basic fuchsin dye. The ground sections were imaged using CLSM in the confocal fluorescence mode. Confocal images revealed that the newly formed bone could be clearly distinguished from bone marrow and cortical bone, as well as from the implant material. PMID- 17117474 TI - In vitro osteoblastic response to 30 vol% hydroxyapatite-polyethylene composite. AB - Hydroxyapatite-reinforced high-density polyethylene (HA-HDPE) composite, as a bone replacement material, has successfully been used clinically as middle ear prostheses and orbital floor implants. The aim of this study was to examine its in vitro biocompatibility in order to develop a further application, that is, as skull reconstruction implants. Human osteoblast cells isolated from femoral heads and crania were used to determine the biological response of the composites. HA HDPE composites (30 vol %) with two grades of HA filler that had different surface morphologies were selected for this in vitro assessment. The results showed that HA-HDPE composite was bioactive and supported osteoblast attachment, proliferation, and differentiation. The composite with rough-surfaced HA filler demonstrated slightly better cellular response than the composite with smooth surfaced HA filler. Although osteoblastic cells derived from skull showed an overall slower response compared with those from femoral heads, these in vitro results show that HA-HDPE composite potentially could be used as a skull implant. PMID- 17117475 TI - Quantification of various growth factors in different demineralized bone matrix preparations. AB - Besides autografts, allografts, and synthetic materials, demineralized bone matrix (DBM) is used for bone defect filling and treatment of non-unions. Different DBM formulations are introduced in clinic since years. However, little is known about the presents and quantities of growth factors in DBM. Aim of the present study was the quantification of eight growth factors important for bone healing in three different "off the shelf" DBM formulations, which are already in human use: DBX putty, Grafton DBM putty, and AlloMatrix putty. All three DBM formulations are produced from human donor tissue but they differ in the substitutes added. From each of the three products 10 different lots were analyzed. Protein was extracted from the samples with Guanidine HCL/EDTA method and human ELISA kits were used for growth factor quantification. Differences between the three different products were seen in total protein contend and the absolute growth factor values but also a large variability between the different lots was found. The order of the growth factors, however, is almost comparable between the materials. In the three investigated materials FGF basic and BMP-4 were not detectable in any analyzed sample. BMP-2 revealed the highest concentration extractable from the samples with approximately 3.6 microg/g tissue without a significant difference between the three DBM formulations. In DBX putty significantly more TGF-beta1 and FGFa were measurable compared to the two other DBMs. IGF-I revealed the significantly highest value in the AlloMatrix and PDGF in Grafton. No differences were accessed for VEGF. Due to the differences in the growth factor concentration between the individual samples, independently from the product formulation, further analyzes are required to optimize the clinical outcome of the used demineralized bone matrix. PMID- 17117476 TI - Osteoblast response to titanium regulates transcriptional activity of Runx2 through MAPK pathway. AB - Bone formation around an implant surface is indispensable for osseointegration of dental implants. Transcriptional factor Runx2 is essential for osteoblast differentiation and bone formation. However, little is known about the involvement of Runx2 in osseointegration. The purpose of this study was to investigate molecular interactions between Runx2 and implant titanium materials at the transcriptional level. Primary osteoblasts isolated from mouse calvaria and C3H10T1/2 cells were cultured on titanium plates or in the presence of titanium particles. Transcriptional activity of Runx2 was measured by reporter assay using osteocalcin gene promoter. We found significant increase (p < 0.05) in Runx2 transcriptional activity in cells cultured on titanium plates compared to plastic plates. Titanium particles also upregulated Runx2 transcriptional activity for over 2-folds compared to control and this effect was abolished by the specific inhibitor for ERK1/2, PD98059. Moreover, treatment with PD98059 clearly suppressed osteoblast mineralization cultured on titanium plates. These data suggest that osteoblast attachment to titanium enhanced Runx2 transcriptional activity and bone formation via MAPK pathway during the osseointegration of dental implants. PMID- 17117477 TI - The role of MAPK in Kupffer cell toll-like receptor (TLR) 2-, TLR4-, and TLR9 mediated signaling following trauma-hemorrhage. AB - Severe injury deranges immune function and increases the risk of sepsis and multiple organ failure. Kupffer cells play a major role in mediating posttraumatic immune responses, in part via different Toll-like receptors (TLR). Although mitogen-activated protein kinases (MAPK) are key elements in the TLR signaling pathway, it remains unclear whether the activation of different MAPK are TLR specific. Male C3H/HeN mice underwent midline laparotomy (i.e., soft tissue injury), hemorrhagic shock (MAP approximately 35 mm Hg for 90 min), and resuscitation. Kupffer cells were isolated 2 h thereafter, lysed and immunoblotted with antibodies to p38, ERK1/2, or JNK proteins. In addition, cells were preincubated with specific inhibitors of p38, ERK1/2, or JNK MAPK followed by stimulation with the TLR2 agonist, zymosan; the TLR4 agonist, LPS; or the TLR9 agonist, CpG DNA. Cytokine (TNF-alpha, interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and KC) production was determined by cytometric bead array after 24 h in culture. MAPK activity as well as TNF-alpha, MCP-1, and KC production by Kupffer cells were significantly increased following trauma-hemorrhage. TLR4 activation by LPS stimulation increased the levels of all measured cytokines. CpG-stimulated TLR9 signaling increased TNF-alpha and IL-6 levels; however, it had no effect on chemokine production. Selective MAPK inhibition demonstrated that chemokine production was mediated via p38 and JNK MAPK activation in TLR2, -4, and -9 signaling. In contrast, TNF-alpha and IL-6 production was differentially regulated by MAPK depending on the TLR pathway stimulated. Thus, Kupffer cell TLR signaling employs different MAPK pathways in eliciting cytokine and chemokine responses following trauma-hemorrhage. PMID- 17117478 TI - Corticotropin releasing factor receptor 1 (CRF1) and CRF2 agonists exert an anti inflammatory effect during the early phase of inflammation suppressing LPS induced TNF-alpha release from macrophages via induction of COX-2 and PGE2. AB - Corticotropin-releasing factor (CRF), the principal regulator of the hypothalamus pituitary-adrenal (HPA) axis, also modulates the inflammatory response directly, via its effect on mast cells and macrophages. On macrophages, it augments production of lipopolysaccharide (LPS)-induced pro-inflammatory cytokines. CRF and its related peptides may also act as anti-inflammatory agents. Aim of the present work was to examine the role of macrophages on the anti-inflammatory effects of CRF-peptides and the mechanism involved. Thus, we examined if CRF receptor 1 (CRF1) and CRF2 agonists exert any anti-inflammatory effect on primary mouse macrophages. We have found that: (a) CRF, Urocortin (UCN)1 and UCN2 transiently suppressed the release of Tumor Necrosis Factor-alpha (TNF-alpha) in LPS-activated macrophages, an effect peaking at 4 h. This effect did not involve changes on TNF-alpha transcription. (b) CRF peptide-induced suppression of TNF alpha release depended on induction of COX-2 and PGE2 synthesis. (c) Use of specific CRF1 and CRF2 antagonists suggested that this effect involved both CRF receptor types. (d) The effect of CRF-peptides on COX-2 was mediated via PI3K and p38MAPK. (e) Longer exposure of macrophages to CRF-peptides resulted in induction of TNF-alpha production via enhancement of its transcription. In conclusion, this is the first report suggesting that CRF1 and CRF2 agonists exert a biphasic effect on macrophages. During the early stages of the inflammatory response, they suppress TNF-alpha release via induction of COX-2/PGE2 while later on they induce TNF-alpha transcription. Hence, the reported anti-inflammatory effect of CRF peptides appears to involve macrophages and is confined at the early stage of inflammation. PMID- 17117479 TI - Regulation of cyclin-dependent kinase 5 and p53 by ERK1/2 pathway in the DNA damage-induced neuronal death. AB - DNA damage is known to be an initiator of neuronal death in neurodegenerative conditions such as Parkinson's and Alzheimer's diseases. The mechanism linking DNA damage and neuronal death is not completely understood. Here, we delineate the mechanism by which neuronal death evoked by DNA damage is controlled. Using mouse cortical neurons and SH-SY5Y human neuroblastoma cells, we identify a critical role of ERK signaling in neuronal death induced by DNA damage upon mitomycin C treatment. In addition, we provide evidence that the ERK signaling regulates Cyclin-dependent kinase 5 (Cdk5) activity and stability of tumor suppressor p53. Mitomycin C increased expression of p35, a specific activator of neuronal Cdk5 in an ERK1/2-dependent manner. Moreover, stability of p53 was increased by its phosphorylation on Ser33 and Ser46 by Cdk5, leading to neuronal death. Finally, we show that activated ERK induced increased expression of the Egr-1 transcription factor, which then bound to the promoter region of p35. We suggest subsequent increase of p35 expression and Cdk5 activity contribute to p53 dependent neuronal death. Thus, the present finding provides a new insight into a molecular mechanism underlying DNA damage-induced neuronal death. PMID- 17117480 TI - The pursuit of creativity in biology. PMID- 17117481 TI - Evolution at the host-retrovirus interface. AB - Retroviruses are unusual amongst animal viruses in their capacity to integrate into host genomes and be transmitted vertically to host progeny. Vertebrate genomes contain numerous and diverse retrovirus-derived sequences reflecting a long co-evolutionary history during which genome invasion has occurred repeatedly, with wide-ranging evolutionary consequences. Over the past 10 years, a detailed picture of retroviral diversity throughout vertebrate genomes has emerged, revealing striking and informative patterns that differ markedly across species. The power of these data to deliver far-reaching insights into the biology and evolution of retroviruses has been significantly advanced by recent studies identifying ongoing genome invasion in wild populations, and by the characterisation of conserved mechanisms of innate antiretroviral defence. PMID- 17117482 TI - Lhx2--decisive role in epithelial stem cell maintenance, or just the "tip of the iceberg"? AB - Stem cell self renewal, maintenance and differentiation are influenced by the convergence of intrinsic cellular signals and extrinsic microenvironmental cues from the surrounding stem cell niche. However, the specific signals involved are often still poorly understood. This is also true for skin epithelial stem cells. Recently, by transcriptionally profiling of embryonic hair progenitors in mice, Rhee et al. have managed to define how murine hair follicle epithelial stem cells are specified and maintained in an undifferentiated state. These authors have identified Lhx2 as a transcription factor functionally positioned downstream of signals necessary to specify hair follicle stem cells such as p63 or NFkappaB, but upstream of signals like Wnt/beta-catenin, Bmp or Shh that are required to drive activated stem cells via the production of transient amplifying cells into terminal differentiation. PMID- 17117483 TI - Early inflammatory arthritis in the rabbit: the influence of intraarticular and systemic corticosteroids on mRNA levels in connective tissues of the knee. AB - OBJECTIVE: Using a rabbit model of inflammatory arthritis, to determine the influence of early disease on expression of specific genes and investigate the influence of intraarticular (IA) and intramuscular (IM) corticosteroids on the regulation of these genes in connective tissues of the rabbit knee. METHODS: Skeletally mature rabbits underwent induction of antigen-induced arthritis or remained untreated as control animals. Four days after disease induction, at an early stage of the disease, animals underwent either IA or IM treatment with glucocorticoids (GC) (5 mg/knee and 10 mg/kg methylprednisolone acetate, respectively). Twenty-four hours following treatment, synovium, menisci, and cartilage of the knee were collected and analyzed for changes in mRNA levels using reverse transcription-polymerase chain reaction for a number of relevant genes: collagen I, collagen II, biglycan, decorin, matrix metalloproteinases-3 and -13 (MMP-3 and MMP-13), cyclooxygenases-1 and -2 (COX-1 and COX-2), tumor necrosis factor-alpha (TNF-alpha), interleukin 1beta (IL-1beta), inducible nitric oxide synthase (iNOS), hyaluronan synthase-2 (HAS-2), and the housekeeping gene beta-actin. RESULTS: Early inflammatory arthritis led to an overall upregulation of most genes assessed, but a downregulation of some genes (iNOS, HAS-2, COX-1) in some tissues. While genes such as collagen II, MMP-3, and MMP-13 were uniformly downregulated by GC treatment in both normal and arthritic tissues, other genes such as collagen I, biglycan, and decorin differed in their pattern of response depending on the tissue examined, the route of drug administration, and whether normal or arthritic tissue was studied. CONCLUSION: Early mRNA changes in RA-like disease led to alterations in all tissues examined. The changes were uniquely altered by GC treatment. Route of GC administration influenced outcome. PMID- 17117484 TI - Establishment of a pilot pediatric registry for chronic vasculitis is both essential and feasible: a Childhood Arthritis and Rheumatology Alliance (CARRA) survey. AB - OBJECTIVE: To identify the need for, and feasibility of, establishing a web-based USA/Canadian registry of children with chronic systemic vasculitis--an otherwise insufficiently studied population. METHODS: Physician members of the Childhood Arthritis and Rheumatology Research Alliance (CARRA; n = 126) were invited to complete 2 surveys exploring vasculitis-related experience, beliefs about childhood versus adult vasculitis, and commitment to contribute patients to a prospective registry. Diagnoses included Wegener's granulomatosis (WG), childhood polyarteritis nodosa, microscopic polyangiitis (MPA), Takayasu's arteritis, primary angiitis of the central nervous system (PACNS), vasculitis, and unclassified vasculitis. RESULTS: One or both surveys were completed by 102 (81%) physicians. Almost half of first-survey respondents had been in practice for > 15 years. Collective estimated lifetime experience was >1500 patients (WG and unclassified vasculitis were the most common diagnoses). Three hundred seventeen children with vasculitis were seen in the year preceding the survey, with most physicians seeing only 2-5 patients. The majority of respondents believed that childhood vasculitis differed from adult disease, particularly with respect to classification criteria and disease activity markers. Fifty-nine members committed to contribute 2 years' data (approximately 120 patients) to a pilot registry limited to time of diagnosis, focusing on WG, MPA, Churg-Strauss syndrome, PACNS, and unclassified vasculitis. CONCLUSION: We obtained overwhelming consensus from an experienced body of pediatric rheumatologists on the need to study childhood-onset vasculitis independently from adult disease, together with commitment from sufficient members to prospectively contribute 2 years' data to a limited pilot registry to answer some basic questions about presenting and diagnostic features and initial treatment practices at disease onset. PMID- 17117485 TI - Occupational exposure to solvents and gender-related risk of systemic sclerosis: a metaanalysis of case-control studies. AB - OBJECTIVE: In 2001 a metaanalysis reported an excess risk of systemic sclerosis (SSc) related to solvents exposure. The magnitude of risk varied among studies and sources of heterogeneity have not been investigated due to a lack of statistical power. We conducted a new metaanalysis to identify features associated with the magnitude of SSc risk in patients exposed to solvents. METHODS: We searched 4 databases (Medline, Pascal, Pascal Biomed, Francis). Inclusion criteria were: case-control study, occupational exposure to solvents (OES) assessed by questionnaire and summarized to "any solvent" or "any organic solvent," SSc defined by the American College of Rheumatology or the consultant's criteria. The quality of studies within this metaanalysis was scored according to the Newcastle-Ottawa scale. Odds ratios (OR) were adjusted for the "publication bias" and validated by a sensitivity analysis. Subgroup analyses investigated the effect of gender, quality of studies, and the type of controls. RESULTS: Among 11 studies (1291 patients and 3435 controls), 9 involved a majority of women (76.2 to 100%), while 2 involved men only. The risk of SSc associated with OES was variable among studies (p for heterogeneity = 0.01) and overrepresentation of higher OR values in smaller studies (p = 0.003) suggested "publication bias." SSc was associated with OES (OR 2.4; 95% CI 1.7-3.4; p < 0.0001), including after adjusting for bias (OR 1.8; 95% CI 1.2-2.5; p = 0.002). The relative risk was higher (p = 0.03) in men (OR 3.0; 95% CI 1.9-4.6; p < 0.0001) than in women (OR 1.8; 95% CI 1.5-2.1; p < 0.0001). CONCLUSION: Whereas SSc affects women predominantly, among subjects with occupational exposure to solvents, men are at higher risk than women for the disease. PMID- 17117486 TI - Disease subsets, antinuclear antibody profile, and clinical features in 127 French and 247 US adult patients with systemic sclerosis. AB - OBJECTIVE: To investigate the specificities of antinuclear antibodies (ANA) associated with systemic sclerosis (SSc) disease classification and internal organ involvement among patients with SSc of different origins (European and American). METHODS: Serum samples from 374 adult patients diagnosed with SSc were studied: 127 French patients (Paris) were compared with 247 US patients (Pittsburgh). Patients were classified into diffuse cutaneous (dc) and limited cutaneous (lc) SSc subsets. Antibodies associated with SSc were determined by protein and/or RNA immunoprecipitation, indirect immunofluorescence, and immunodiffusion. RESULTS: SSc classification differed significantly in the 2 cohorts: lcSSc and overlap patients with lcSSc combined made up 76% of the French series versus 52% of the US group (p < 0.0001). The frequency of anti-RNA polymerase III antibody was significantly increased in US patients compared with French patients (p < 0.0001). The frequency of anti-topoisomerase I (topo I) antibody was significantly increased among French patients (p < 0.0048). Anti topo I-positive French SSc patients were less likely to have dcSSc (38% vs 65%) and more likely to have milder disease than US anti-topo I-positive patients. The French dcSSc patients had lower proportions of joint/tendon manifestations and renal crisis (7% vs 17%), but more often had radiographic evidence of pulmonary fibrosis (57% vs 30%). French lcSSc patients had a lower frequency of pulmonary arterial hypertension than US lcSSc patients (9% vs 31%; p = 0.002). CONCLUSION: There are disease classification and SSc-related serum autoantibody differences between French and American patients with SSc. These differences help to explain variations in clinical features reported from different geographic regions. PMID- 17117487 TI - Elevated gene expression of Th1/Th2 associated transcription factors is correlated with disease activity in patients with systemic lupus erythematosus. AB - OBJECTIVE: T-box expressed in T cells (T-bet) and GATA-binding protein 3 (GATA-3) are transcriptional factors that play a crucial role in Th1 and Th2 development. We investigated the immunomodulatory roles of T-bet and GATA-3 and Th1/Th2 related cytokines in the pathogenesis of systemic lupus erythematosus (SLE) and their association with disease activity. METHODS: Gene expressions of T-bet, GATA 3, interferon-gamma (IFN-gamma), and interleukin 4 (IL-4) in peripheral blood mononuclear cells, and plasma concentrations of the Th1/Th2 cytokines IFN-gamma, IL-18, and IL-4, were assayed in 80 patients with SLE and 40 sex and age matched healthy subjects by real-time quantitative polymerase chain reaction and ELISA. RESULTS: The mRNA levels of T-bet and IFN-gamma and the relative expression levels of T-bet/GATA-3 and IFN-gamma/IL-4 were significantly higher, in contrast to the lower expressions of GATA-3 and IL-4, in SLE patients than controls (all p < 0.05). In all SLE patients, there were significant correlations in mRNA expression of T-bet with IFN-gamma (r = 0.590, p < 0.0001), and of GATA-3 with IL 4 (r = 0.245, p = 0.029). The relative expressions of T-bet/GATA-3 and IFN gamma/IL-4 correlated with lupus disease activity (r = 0.229, p = 0.042; r = 0.231, p = 0.040, respectively). Plasma IL-18 concentration was increased significantly in all SLE patients (p < 0.05). The elevated plasma Th1/Th2 cytokine ratio IL-18/IL-4 correlated positively with disease activity in all SLE patients (r = 0.250, p = 0.025). CONCLUSION: There is an association between expression of Th1/Th2 transcription factors and cytokines in SLE. The elevated gene expressions of Th1/Th2 transcription factors and cytokines should provide a useful tool for assessing the functional status of T-helper lymphocytes in SLE disease development. PMID- 17117488 TI - Atherosclerotic vascular events in a single large lupus cohort: prevalence and risk factors. AB - OBJECTIVE: To determine prevalence and type of atherosclerotic vascular events (AVE) occurring after entry to the University of Toronto Lupus Clinic; and to compare risk factors in patients with systemic lupus erythematosus (SLE) with AVE to matched SLE controls without AVE. METHODS: Patients with SLE attending the University of Toronto Lupus Clinic who did not have AVE prior to clinic entry were included. Patients have been followed at 2-6 months since 1970 according to a standard protocol. Cases with AVE were matched for sex, era at first clinic visit (1970s, 1980s, 1990s +), inception status, age at first visit, and duration of followup. Chi-square, Fisher's exact, paired T test, and McNemar test were used. Comparison of risk factors for the development of AVE was done using a stepwise conditional logistic regression model for matched pairs. RESULTS: In a total cohort of 1087 SLE patients followed from 1970 until 2004, the prevalence of AVE was 10.9%, and in 561 inception patients it was 9.6%. In multivariate analyses, neuropsychiatric involvement was significantly associated with AVE in both the total and inception cohorts. Smoking was also associated with AVE in the inception cohort, whereas the number of coronary artery disease (CAD) risk factors and vasculitis were significant in the total cohort. CONCLUSION: AVE are major contributors to the clinical presentation of late-stage lupus. A combination of lupus related factors and classic CAD risk factors contributed to the development of AVE. PMID- 17117489 TI - A comparative study of patient-reported functional outcomes in acute rheumatoid arthritis. AB - OBJECTIVE: To assess the performance of the Barthel Index (BI) in patients with rheumatoid arthritis (RA) in the acute care hospital, as compared to the Stanford Health Assessment Questionnaire (HAQ) and the Hannover Functional Questionnaire (Funktionsfragebogen Hannover, FFbH). METHODS: A prospective study of 97 patients with RA admitted to an acute rheumatology hospital with new onset or acute flare of RA. Patients were required to self-complete the BI, the HAQ, and the FFbH. Disease activity was measured using the Disease Activity Score (DAS28). RESULTS: Seventy-eight percent of patients were female, average age was 61.5 (SD 12.5) years, and 72.2% were rheumatoid factor-positive. The median HAQ was 1.29 (range 0-3), median FFbH was 50% (6-100%), and median BI was 95 (0-100), and distribution was highly skewed. All measures of physical functioning were significantly correlated with each other and with the DAS28; however, the BI discriminated poorly between low and high disease activity. CONCLUSION: The BI is not a useful instrument to assess physical functioning in patients with acute symptoms of RA, but may have a role in assessing patients with comorbidities and in assessing nursing care needs in the acute care hospital. PMID- 17117490 TI - Rheumatoid arthritis and spondyloarthropathies: geographical variations in prevalence in France. AB - OBJECTIVE: To determine geographical variation in the prevalence of rheumatoid arthritis (RA) and spondyloarthropathies (SpA) in France. METHODS: The survey sample was drawn from 7 areas of France. Households were randomly selected using the national telephone directory, and an individual within each household was randomly chosen by the next-birthday method. All cases of suspected RA and SpA were confirmed by the patient's rheumatologist or by clinical examination. Standardized estimates of prevalence were compared between regions and groups of regions. RESULTS: In total 15,219 anonymous telephone numbers were selected. An average response rate of 64% led to a total of 9395 respondents included in the study. The highest regional rates of RA were observed in the south (range 0.59 0.66%), and the lowest in the north (range 0.14-0.24%), with a national rate of 0.31% (95% CI 0.18-0.48%). Regional heterogeneity was observed for SpA, with the highest rates in Bretagne (0.47%) and the Sud-Est (0.53%) and a national rate of 0.30% (95% CI 0.17-0.46%). CONCLUSION: This study is the largest of its kind conducted in France. It shows inter-regional variations, mainly in RA, with a higher prevalence in the south of the country. The many potential reasons for the heterogeneity observed, including genetic and environmental factors, warrant further research. PMID- 17117491 TI - Lymphoproliferative disorders in rheumatoid arthritis: clinicopathological analysis of 76 cases in relation to methotrexate medication. AB - OBJECTIVE: Individuals with rheumatoid arthritis (RA) with or without methotrexate (MTX) medication occasionally develop lymphoproliferative disorders (MTX-LPD and non-MTX-LPD, respectively). The hyperimmune state of RA itself or the immunosuppressive state induced by MTX administration might contribute to development of LPD. Our objective was to characterize MTX-LPD in comparison to non-MTX-LPD and sporadic LPD in patients with RA. METHODS: We compared MTX-LPD to non-MTX-LPD and sporadic LPD by evaluating 48 cases of MTX-LPD, 28 non-MTX-LPD, and 150 sporadic LPD. RESULTS: Later onset age of LPD and female predominance were evident in patients with RA-LPD compared to sporadic LPD. The interval between the diagnosis of RA and LPD in MTX-LPD (median 132 mo) was significantly shorter than that in non-MTX-LPD (240 mo). The frequency of diffuse large B cell lymphoma (DLBCL) and positive rate of Epstein-Barr virus (EBV) in RA-LPD was significantly higher than in sporadic LPD (57.9% vs 42.7%, 27.6% vs 9.9%, respectively). After withdrawal of MTX, 11 of the MTX-LPD cases showed a spontaneous regression of tumors. The 5-year survival rate in RA-LPD (59.2%) was significantly worse than that in sporadic LPD (74.6%). CONCLUSION: The majority of cases of RA-LPD show similar clinicopathological characteristics irrespective of MTX medication, except for spontaneous regression of LPD after withdrawal of MTX in MTX-LPD, and a shorter interval between the diagnosis of RA and LPD in MTX LPD than in non-MTX-LPD. RA-LPD cases showed younger age of onset, female predominance, unfavorable prognosis, and higher frequencies of DLBCL and EBV positivity compared to sporadic LPD. PMID- 17117492 TI - Risk factors for surgical site infections and other complications in elective surgery in patients with rheumatoid arthritis with special attention for anti tumor necrosis factor: a large retrospective study. AB - OBJECTIVE: To identify risk factors for surgical site infection (SSI) in patients with rheumatoid arthritis (RA) with special attention for anti-tumor necrosis factor (anti-TNF) treatment. METHODS: All patients with RA who had undergone elective orthopedic surgery since introduction of anti-TNF were included in a retrospective parallel-cohort study with a one-year followup. Primary endpoint was a SSI according to the 1992 Centers for Disease Control and Prevention criteria and/or antibiotic use. Cohort 1 did not use anti-TNF, cohort 2 used anti TNF but had either stopped (2A) or continued anti-TNF preoperatively (2B), the cutoff point being set at 4 times the half-life time of the drug. Infection rates were compared between cohorts, and logistic regression analysis was performed to examine risk factors. RESULTS: In total, 1219 (768 patients) procedures were included, and crude infection risks were 4.0% (41/1023), 5.8% (6/104), and 8.7% (8/92) in cohorts 1, 2A, and 2B, respectively. Elbow surgery (OR 4.1, 95% CI 1.6 10.1), foot/ankle surgery (OR 3.2, 95% CI 1.6-6.5), and prior skin or wound infection (OR 13.8, 95% CI 5.2-36.7) were associated with increased risk of SSI, whereas duration of surgery (OR 0.42, 95% CI 0.23-0.78) and sulfasalazine use (OR 0.21, 95% CI 0.05-0.89) were associated with decreased risk. Perioperative use of anti-TNF was not significantly associated with an increase in SSI rates (OR 1.5, 95% CI 0.43-5.2). CONCLUSION: The most important risk factor for SSI is history of SSI or skin infection. Although our study was not powered to detect small differences in infection rates, perioperative continuation of anti-TNF does not seem to be an important risk factor for SSI. PMID- 17117493 TI - Implementing Gaussian process inference with neural networks. AB - Gaussian processes compare favourably with backpropagation neural networks as a tool for regression, and Bayesian neural networks have Gaussian process behaviour when the number of hidden neurons tends to infinity. We describe a simple recurrent neural network with connection weights trained by one-shot Hebbian learning. This network amounts to a dynamical system which relaxes to a stable state in which it generates predictions identical to those of Gaussian process regression. In effect an infinite number of hidden units in a feed-forward architecture can be replaced by a merely finite number, together with recurrent connections. PMID- 17117494 TI - On the universal clustering under a broad class of loss functions. AB - Determination of the number of significant clusters in the sample represents a very important problem. It is expected that the outcome of clustering under a broad class of loss functions will be more stable if the correct number of clusters is used. In order to illustrate the model of universal clustering we consider 1) family of power loss functions in probabilistic space; 2) family of exponential loss functions in Euclidean space. The proposed model is general, and proved to be effective in application to the synthetic datasets. PMID- 17117495 TI - Regularization network-based gene selection for microarray data analysis. AB - Microarray data contains a large number of genes (usually more than 1000) and a relatively small number of samples (usually fewer than 100). This presents problems to discriminant analysis of microarray data. One way to alleviate the problem is to reduce dimensionality of data by selecting important genes to the discriminant problem. Gene selection can be cast as a feature selection problem in the context of pattern classification. Feature selection approaches are broadly grouped into filter methods and wrapper methods. The wrapper method outperforms the filter method but at the cost of more intensive computation. In the present study, we proposed a wrapper-like gene selection algorithm based on the Regularization Network. Compared with classical wrapper method, the computational costs in our gene selection algorithm is significantly reduced, because the evaluation criterion we proposed does not demand repeated training in the leave-one-out procedure. PMID- 17117496 TI - Mixture models for detecting differentially expressed genes in microarrays. AB - An important and common problem in microarray experiments is the detection of genes that are differentially expressed in a given number of classes. As this problem concerns the selection of significant genes from a large pool of candidate genes, it needs to be carried out within the framework of multiple hypothesis testing. In this paper, we focus on the use of mixture models to handle the multiplicity issue. With this approach, a measure of the local FDR (false discovery rate) is provided for each gene. An attractive feature of the mixture model approach is that it provides a framework for the estimation of the prior probability that a gene is not differentially expressed, and this probability can subsequently be used in forming a decision rule. The rule can also be formed to take the false negative rate into account. We apply this approach to a well-known publicly available data set on breast cancer, and discuss our findings with reference to other approaches. PMID- 17117497 TI - The prediction of bacterial transcription start sites using SVMs. AB - Identifying promoters is the key to understanding gene expression in bacteria. Promoters lie in tightly constrained positions relative to the transcription start site (TSS). In this paper, we address the problem of predicting transcription start sites in Escherichia coli. Knowing the TSS position, one can then predict the promoter position to within a few base pairs, and vice versa. The accepted method for promoter prediction is to use a pair of position weight matrices (PWMs), which define conserved motifs at the sigma-factor binding site. However this method is known to result in a large number of false positive predictions, thereby limiting its usefulness to the experimental biologist. We adopt an alternative approach based on the Support Vector Machine (SVM) using a modified mismatch spectrum kernel. Our modifications involve tagging the motifs with their location, and selectively pruning the feature set. We quantify the performance of several SVM models and a PWM model using a performance metric of area under the detection-error tradeoff (DET) curve. SVM models are shown to outperform the PWM on a biologically realistic TSS prediction task. We also describe a more broadly applicable peak scoring technique which reduces the number of false positive predictions, greatly enhancing the utility of our results. PMID- 17117498 TI - Value at risk estimation using independent component analysis-generalized autoregressive conditional heteroscedasticity (ICA-GARCH) models. AB - We suggest using independent component analysis (ICA) to decompose multivariate time series into statistically independent time series. Then, we propose to use ICA-GARCH models which are computationally efficient to estimate the multivariate volatilities. The experimental results show that the ICA-GARCH models are more effective than existing methods, including DCC, PCA-GARCH, and EWMA. We also apply the proposed models to compute value at risk (VaR) for risk management applications. The backtesting and the out-of-sample tests validate the performance of ICA-GARCH models for value at risk estimation. PMID- 17117499 TI - Manifold learning for robot navigation. AB - In this paper we introduce methods to build a SOM that can be used as an isometric map for mobile robots. That is, given a dataset of sensor readings collected at points uniformly distributed with respect to the ground, we wish to build a SOM whose neurons (prototype vectors in sensor space) correspond to points uniformly distributed on the ground. Manifold learning techniques have already been used for dimensionality reduction of sensor space in navigation systems. Our focus is on the isometric property of the SOM. For reliable path planning and information sharing between several robots, it is desirable that the robots build an internal representation of the sensor manifold, a map, that is isometric with the environment. We show experimentally that standard Non-Linear Dimensionality Reduction (NLDR) algorithms do not provide isometric maps for range data and bearing data. However, the auxiliary low dimensional manifolds created can be used to improve the distribution of the neurons of a SOM (that is, make the neurons more evenly distributed with respect to the ground). We also describe a method to create an isometric map from a sensor readings collected along a polygonal line random walk. PMID- 17117501 TI - Help tips. Flossing your teeth. PMID- 17117500 TI - The national sleep debt. Too many get too little. PMID- 17117502 TI - Kidney stone treatment may have delayed risks. PMID- 17117503 TI - Burning mouth syndrome. Turning down the heat. PMID- 17117504 TI - Jet lag. Getting back in sync. PMID- 17117505 TI - My doctor recently discovered that I have a small umbilical hernia. It's never bothered me, but I was a little surprised when he suggested we leave it as is unless it gives me any trouble. I thought hernias always had to be repaired. Is this something new? PMID- 17117506 TI - What are bioidentical hormones? Natural. Bioidentical. Compounded. Confusion about these terms is only adding to the confusion over hormone therapy. PMID- 17117507 TI - Managing atrial fibrillation. When heart rhythm is in disarray, medications can often help. but if they don't do the trick, other treatments are available. PMID- 17117508 TI - Breath control helps quell errant stress response. Breathe in deeply to a count of four and out again slowly. There. Don't you feel calmer? PMID- 17117509 TI - By the way, doctor. I'm 48 years old, and 26 years ago I had a cone biopsy after an abnormal Pap test. Since then, all my Pap tests have been normal. But for the past three years, scar tissue has made it difficult for my gynecologist to get adequate cells. What do I do about Pap smears now? PMID- 17117510 TI - [Congenital absence of the ductus venosus associated with severe congenital heart malformation--case report and review of the literature]. AB - Congenital absence of the ductus venosus is a rare anomaly. The authors have found 57 cases in the literature. Their case was associated with a complex congenital heart disease, what occurs only in 10% of absent ductus venosus cases. In cases, where the umbilical vein connects directly to the right atrium, the volume overloading causes dilatation of the right atrium and ventricle, development of polyhydramnios and fetal hydrops. The diagnosed an umbilical vein directly connected to the right atrium, extreme systemic congestion, but no hydrops. In some fetuses this condition did not result hydrops, because there is compensation by the rich compliance of intrahepatic vascular beds. This case is an illustration that mild sonographic signs such a transient bradycardia in the early second trimester could be a sign of a heart malformation and can draw attention of the sonographer to other associated severe congenital heart disease. PMID- 17117511 TI - [New challenges and opportunities in a globalized world: a political analysis of the response to HIV/AIDS in Mexico]. AB - This work is a synthesis of an investigation into the political economics of HIV/AIDS in Mexico within the context of globalization. It is based on a distinction between what is defined as neo-liberal globalism (an ideological and overwhelmingly economic agenda) and globalization (a worldwide, multidimensional phenomenon). The core argument is that the existence of real alternatives to the neo-liberal paradigm results from a series of political, economic and social circumstances which are very real in Mexico today in the context of globalization, which have given rise to a rather more inclusive policy-making process. Special emphasis is given to the contrast between exclusive public policy systems, such as those seen in the process of structural economic reforms or the re-structuring of the financing of healthcare systems, and public policy systems which work with HIV/AIDS, which tend to be more inclusive and democratic. Both types of system operate in Mexico and in other parts of the world. PMID- 17117512 TI - [The national-international nexus in public health: Uruguay and the circulation of child health and welfare policies, 1890-1940]. AB - This paper examines the emergence of child health policy as a global concern from the perspective of Uruguay's interaction with the international public health community in the early 20th century and its role in the circulation of child health ideas and practices. It considers two facets of national-international interplay: a) Uruguay's exploration of an interrelationship with the international panorama of policies, research and lessons on the improvement of child health; and b) the translation of Uruguay's domestic debates into the influential Instituto Internacional Americano de Proteccion a la Infancia - IIPI and the purveying of Uruguay's policies, research, and lessons on child health improvement to other countries and to international health and welfare agencies. PMID- 17117513 TI - Sick city: maps and mortality in the time of cholera. PMID- 17117514 TI - [Public health in Cuba and its international experience (1959-2005)]. AB - The past 42 years of Cuba's healthcare system have been characterized, among other things, by broad-based international experience and projects, which began in 1962 with the deployment of the first Cuban medical mission to Algeria. Since then, it has collaborated with more than 101 countries in Asia, Africa and Latin America, providing aid missions to areas suffering from natural disasters and sending medical staff to regions where they did not exist before. It has also helped some countries draw up medical development plans according to their specific needs. Cooperation with human resource training and development has also been carried out, with the result that hundreds of thousands of youths have been able to start their medical studies in countries all over the Americas and in some nations from Asia and Africa. PMID- 17117515 TI - [Reluctant partner: Canada's relationship with the Pan American Health Organization (PAHO)]. AB - Despite a strong commitment to multilateralism and international health cooperation in the post World War II era, Canada refrained form joining the Pan American Health Organization - PAHO until 1971. Drawing on letters and memos sent between Canadian diplomats and government representatives, this paper explores official Canadian accounts of the factors that delayed Canada's membership in PAHO. These factors include the initial lack of official relations between Canada and Latin America, US hegemony in the region, and budgetary constraints. Canada's cautious position regarding PAHO is also placed within the context of Canada's overall foreign policy to the region, emphasizing the parallels between Canada's reluctant association with PAHO and the evolution of Canada's engagement with the region as a whole. PMID- 17117516 TI - Striking the right balance. PMID- 17117517 TI - [Open circuit: the exchange of medical and scientific knowledge in Latin American in the early 20th century]. AB - This article discusses the Latin American Medical Congresses and International Exhibitions on Hygiene held in the first few decades of the 20th century as a strategy for underpinning and influencing medical knowledge within the specialized community itself and for public authorities, which were fundamental for presenting to society at large as they were seen as the vehicles of official know-how on the art of medicating. These events made up part of a broader movement to internationalize and coordinate the professional field of medicine in Latin America. The article further suggests that the activities that took place during these events played a key role in the propagation of ideas and exchange of experience between Latin American nations, forming a network of scientific exchange in the continent. PMID- 17117518 TI - The trouble with making combination drugs. PMID- 17117519 TI - Study reveals secrets to faster drug development. PMID- 17117520 TI - An audience with... Sir Alasdair Breckenridge. Interview by Joanna Owens. PMID- 17117521 TI - Making gains in pain. PMID- 17117522 TI - Decitabine. PMID- 17117523 TI - [Elizabeth Fee: a historian reaching out to wider audiences]. AB - To what activities and topics does a historian in health and medicine, whose articles and books have become fundamental references for scholars of the area, devote her time? Feminism, counter-culture, medical education, global health, the role of international health organizations, and knowledge sharing in the health history are some of the subjects Elizabeth Fee addresses in this interview given at Fiocruz in April where she presented the 2006 inaugural class to the Graduate Program in History of Health Sciences at Casa de Oswaldo Cruz. The topic of her lecture was "The World Health Organization and AIDS: what can we learn from history?" PMID- 17117524 TI - New diagnostic technology: applications in animal health and biologics controls. Proceedings of an international conference. October 3-5, 2005. Saint-Malo, France. PMID- 17117525 TI - [Home oxygen inhalation therapy--preparation for electricity blackout]. PMID- 17117526 TI - [Na+/Ca+ exchanger]. PMID- 17117527 TI - [Cell adhesion molecules]. PMID- 17117528 TI - [Sex difference in pharmacokinetics]. PMID- 17117529 TI - [Home oxygen inhalation therapy during flight]. PMID- 17117530 TI - [Conditional stimulus or conditional reinforcer? The current trend in the renewed study on drug dependence]. PMID- 17117531 TI - [Various problems in patient compliance for home oxygen inhalation therapy]. PMID- 17117532 TI - Catholic sisters and nursing in the Civil War. PMID- 17117533 TI - Ruthless Rufus. PMID- 17117534 TI - Ready to die. PMID- 17117535 TI - Brain death and organ transplantation: knowledge, attitudes, and practice among Japanese students. AB - OBJECTIVE: To investigate the knowledge, attitudes and practice of Japanese students regarding brain death and organ transplantation. METHODS: A 22-item questionnaire was handed out among 383 Japanese students during the 2002 academic year. The data was finally analyzed using a statistical package for social sciences, SPSS. RESULTS: Most students knew that organ transplantation can save a life 97%, while only 38% of the students were aware that there is no treatment for brain dead patients. Overall, 60% of the respondents believed brain death is equal to human death and 63% believed that organ removal from brain dead donors is mutilation of the body. There was a negative association between the students who reported positive attitudes toward the idea that removing an organ is mutilation of the body with their willingness to be a donor. Of the students 27% were opposed to the necessity of family agreement for organ donation. Although 66% expressed their wish to be a donor, less than half of them, 31% had a donor card; and 40% did not even know how to get a donor card. CONCLUSION: Granting a veto power to the family as well as the mistaken idea that there is recovery from brain death, can be considered as obstacles to organ procurement from the brain dead in Japan. The high value of life among Japanese society and the willingness to be a donor, bring hope that giving correct medical information and proper public education can increase social acceptance of organ transplantation from the brain dead in Japan. PMID- 17117536 TI - Commentary on Bagheri et al. PMID- 17117537 TI - Compromised autonomy, and Asian autonomy: commentaries on Glock & Goldim, and Dena Hsin-Chen Hsin. PMID- 17117538 TI - The limitations of the Dutch concept of euthanasia. AB - Why has the law on euthanasia in the Netherlands caused such an upheaval both at home and abroad? In this paper I explore some bioethical issues in the decriminalisation of euthanasia in the Netherlands. The regulatory role of legal and state institutions in the process of decision-making by patients, physicians and other people concerned plays a central role in these discussions. I argue, first, that the limited scope of the Dutch legislation on euthanasia cannot be a solution to end-of-life issues in general, and, second, that it is inadequate as a model for dealing with problems related to 'death-on-request' abroad. Moreover, the confusion around the meaning of the term euthanasia would make its adaptation in other institutional environments capricious. Legal changes in the Netherlands was accompanied by cultural changes, expressed in the use of terms such as individual autonomy and personal choice. In the last section of this article I argue that the social and political environment may be crucial in defining the meaning of free choice. The contending views on the decriminalisation of euthanasia seem to develop as a reaction to change in medical technology in a particular socio-political environment rather than from a unique cultural ethic. PMID- 17117539 TI - When did "bioethics" begin in each country? A proposal of a comparative study. PMID- 17117540 TI - Autonomy as a universal expectation: a review and a research proposal. AB - In the World Health Report 2000 WHO introduces ethical issues in the elevation of health systems responsiveness performance. Although we consider this a positive step, the parameters considered in the Report are in some cases unsustained by extensive research. This is the case of autonomy, which is postulated as a "universal expectation". As we think that this is culture-linked issue we argue that such kind of universal categorizations lacks substantive empirical evidence. We undertook a short review of a small intentional sample of international literature, in order to demonstrate that regardless of the philosophical status of autonomy as a principle or category there is not background enough to postulate it as a universal expectation. We propose international collaborative research to define the issue, using qualitative research methodology. PMID- 17117541 TI - Genetic health care services, present and near future in Japan. AB - A critical review from standpoints of ethical, legal and social implications (ELSI) is required to properly develop and distribute genetic services, including genetic testing. In addition to assure analytical validity, clinical validity and clinical utility of the testing, public opinion and perceptions of the community regarding implementation of the testing are important for obtaining informed consent prior to testing. Otherwise, the testing could be done on only limited numbers of community members. Development of effective inversion in clinical practices and regulation to provide confidentiality of all genetic information should bolster the attitude of the community. Education of the public and mass media is the most important factor if progress is to be made in the clinical application of human genome research, and to avoid "genohype". PMID- 17117542 TI - Assisted reproduction--Islamic views on the science of procreation. AB - The field of assisted reproduction is one of the fastest growing areas in medicine. The development of new and sophisticated techniques all aim to give reproductive technologies and scientists a better understanding of reproductive biology. The development of new techniques may also be able to improve the chances of an infertile couple towards achieving a pregnancy. However the ethics and legality of assisted reproductive technology must not be brushed aside. Islamic scholars whose reference basis resides in the Holy Quran and the Hadith have addressed different issues relating to assisted reproductive technology. Some of the issues remain controversial and need detailed discussion before it can be deemed as appropriate and legible for use as guidelines in the development of the processes involved. PMID- 17117543 TI - ["The art of killing by electricity": the sublime and the electric chair]. PMID- 17117544 TI - Women's voluntarism, special education, and the Junior League: "social motherhood" in Atlanta, 1916-1968. PMID- 17117545 TI - [Monitoring during oxygen inhalation therapy--transcutaneous PCO2 measurement system]. PMID- 17117546 TI - War, region, and social welfare: federal aid to servicemen's dependents in the South, 1917-1921. PMID- 17117547 TI - [Prevention of neoplasm--focus point of study of prevention and therapy of neoplasm in 21st century]. PMID- 17117548 TI - [Galanin increases the survival rate of hippocampal cells injured by H2O2 in vitro]. AB - The method of primary hippocampal nerve cell culture was used to study the injury effect of H2O2 and the protective effect of galanin (GAL) and GAL receptor agonists. Result demonstrated that H2O2 has obvious dose relative toxicity to hippocampal cells in vitro. GAL and GAL's nonselective agonist GAL1-11, GalR2's selective agonist GAL2-11 can increase the survival rate of hippocampal cells suffered form H2O2. All the protective effects can be blocked by nonselective antagonist M35. The result indicates that GAL can protect hippocampal cells from oxidative injury in vitro, which is most probably mediated by GalR2. PMID- 17117549 TI - [Comparative proteome analysis of hepatoma cells at subcellular level]. AB - The subcellular proteome strategy can complement the separation power of two dimensional electrophoresis, a step for subcellular fractionation is introduced before electrophoresis is conducted, and more proteins will be displayed in two dimensional gels. A comparative analysis of proteomic profiling of mitochondria and nuclei was conducted between hepatoma cell and hepatocyte, in order to find more information involved in cancer development. The cultured hepatoma cell line QGY-7703 and hepatocyte line LO2 were used as research models in this work. Subcellular fractionation for mitochondrion and nucleus were done by ultracentrifugation, then two-dimensional electrophoresis was applied to the separation of mitochondrial and nuclear proteins, imaging analysis for the selection of differentially-expressed protein spots, and MALDI-TOF-MS for the identification of proteins. 54 spots from electrophoresis gels were selected as differentially-expressed protein for MS analysis which resulted in identification for 22 proteins. Among the 22 differentially-expressed proteins, 17 show a up regulated expression and 5 show a down-regulated expression. These differentially expressed proteins found in this work have a wide coverage of functions which are related to energy metabolism, cytosheleton, protein biosynthesis, pre-mRNA splicing and processing, apoptosis regulation. These results imply that cancer cell has experienced a fundamental change in structure and metabolism pattern. PMID- 17117550 TI - [Proteomic approach to the effect of epirubicin on hepatoma cells at subcellular level]. AB - Epirubicin is an antineoplastic agent known as an anthracycline. It acts directly on DNA by blocking its replication and transcription, so apoptosis can be induced for cancer cells. The protein expression of cancer cells will be altered due to the induction of pharmacological action of epirubicin, so it is important to pay attention to the altered profiling of proteins. Here proteomic strategy was applied to the hepatoma cells at subcellular level, comparative proteome analysis of mitochondria and nucleus were conducted between the hepatoma cells administered with epirubicin and the not-administered. Centrifugation was used for the subcellular fractionation, then 2-DE for the separation of proteins, imaging analysis for the diction of expression-altered spots, and MALDI-TOF-MS for the identification of proteins. In total, 15 proteins were found to have altered their expression after the induction of epirubicin, among them, 5 proteins showed up-regulated expression and 10 showed down-regulated expression. These altered proteins are involved in life processes of cells such as energy metabolism, protein biosynthesis, structure of cell skeleton, processing and maturation of mRNA, heat shock of cells and apoptosis. PMID- 17117551 TI - [Construction and immunogenicity of hCG chimeric peptides (CP1 and CP10) containing three self- B cell epitopes and six foreign T cell epitopes]. AB - Two bio-synthesizing chimeric peptide (CP) immunogens named CP1 and CP10 have been designed, which consist of three linear B cell epitopes (BCE) of human chorionic gonadotropin beta subunit (beta-hCG) and six foreign T cell epitopes including two "promiscuous" TCEs from hepatitis B surface antigen and tetanus toxoid. Two artificial genes encoding CP1 and its derivative CP10 were synthesized, which could be expressed in E. coli at the level of about 1% of the total cell proteins when inserted into the thermo-induction vector respectively. In Western blot tests, the expressed CP1 and CP10 proteins with about 16.5 kD shown on the SDS-polyacrylamide gel electrophoresis (PAGE) gel can be recognized by the monoclonal or polyclonal antibodies specific to each linear epitope of beta-hCG. Each of expressed proteins can be purified with 95% relative homogeneity using our improved method of preparative gel PAGE. Their yields were about 1-2 mg per 12 L culture. Also, the CPl and CP10 immunogens can induce antibodies in mice that recognize recombinant CP1 betar CP10 and natural beta hCG, and there are three anti-beta5, beta9 and beta8 BCE antibodies in their antisera. The construction and expression of beta-hCG CP1 and CP10 will provide new immunogens for developing an ideal and superior hCG birth control and/or tumor therapeutic vaccine. PMID- 17117552 TI - [Study on embryo development and somatic embryogenesis of jujube]. AB - The results of embryological observation on embryo development in the thirteen cultivars of jujube indicated that they shared the same experience from proembryo to globular embryo, heart-shaped embryo, torpedo-shaped embryo and cotyledon stage embryo. Embryo began aborting from globular or heart-shaped embryo period in most of cultivars of jujube, but in few of them embryo could keep synchronous developing to cotyledon stage embryo. And the courses of embryo development were not synchronous. PMID- 17117553 TI - [The study of thrombospondin-I (TSP1) expression in the early stages of diabetic retinopathy induced by streptozotocin]. AB - Thrombospondin1 (TSP1) is a known antiangiogenic factor, but its possible alteration during the early stages of diabetic retinopathy has not been explored yet. The present study sought to investigate the expression of TSP1 using a model of streptozotocin (STZ) induced diabetes in the rat. Diabetes was induced in male SD rats by an intraperitoneal injection of STZ. Age matched animals served as control. 2 months after the onset of diabetes, the expression of TSP1 mRNA was quantified in retinal tissue by quantitative real time reverse transcription polymerase chain reaction (RT-PCR) respectively. The locations of TSP1 normal and diabetic retinas were also established by immunohistochemistry. 2 months after the onset of diabetes, retinal TSP1 concentrations were significantly increased in the diabetic rats compared with control rats. Diabetes caused the upregulation of TSP1 expression in the layers of the retina 2 months after the induction of diabetes. The upregulation of TSP1 in retinas of diabetic rats suggests a role in experimental diabetic retinopathy. Further studies should address the possible involvement of the TSP1 in the pathophysiology of diabetic retinopathy. PMID- 17117554 TI - [Isoflavone accumulation associated with cell structural changes in Maackia amurensis suspension cultures elicited by methyl jasmonate, salicylic acid and nitric oxide]. AB - Comparison between changes in isoflavone accumulation and cell structure in Maackia amurensis suspension cultures elicited by methyl jasmonate (MeJA),salicylic acid (SA) and nitric oxide (NO) was studied. The results suggested that MeJA, SA and NO can all stimulate isoflavone production remarkably. After 9d treatment with 200 micromol/L MeJA,100 micromol/L SA and 50 micromol/L SNP, the isoflavone content was 417.18%, 185.45% and 222.45% of the control, respectively. At the same time, the electron-dense body (EDB) could be easily found in the cells, while its number achieved most after 9 d treatment,and the number of EDB and isoflavone content were positive correlated. It was deduced that elicitors stimulated plant secondary metabolites production associated with changes in cell structure. PMID- 17117555 TI - [Tissue culture and high-frequency plant regeneration of buckwheat (Fagopyrum esculentum Moench)]. AB - Different explants and compounding proportions of different hormones were comparatively studied in tissue culture of Fagopyrum esculentum Moench and thereafter an efficient plant-regeneration system was established by in vitro F. esculentum Moench culture. On MS medium containing 2.0 mg/L 2,4-D and 1.0 mg/L 6 BA, 89.6% cotyledon segments could be induced to produce calli; however, on MS medium containing 2.0 mg/L 2,4-D and 1.0-2.0 mg/L 6-BA, the induction rate of hypocotyl segments could reach as high as 100%. On MS medium containing 2.0 mg/L 6-BA, 0.1 mg/L IAA and 1 mg/L KT, adventitious buds could be regenerated indirectly from calli or directly from explants, the differentiation rates of cotyledon-derived calli and hypocotyl-derived calli were 42.5% and 73.6% respectively; the calli coming from hypocotyl segments differentiated evidently at a higher rate than the calli originated from cotyledon segments. Well-grown adventitious buds were inoculated on 1/2 MS medium containing 1.0 mg/L IBA and 0.5 mg/L NAA, 100% rooting frequency was obtained. Plantlets grew well and appeared normal with no mortality after being transplanted to soil. Moreover, the survival rate of plantlets reached 91.6%. PMID- 17117556 TI - [Purification and characterization of hemolymph lectin from Tegillarca granosa Linnaeus]. AB - Agglutinin with hemagglutinating activity was found in hemolymph of Tegillarca granosa Linnaeus. By extraction, fraction with saturated ammonium sulfate precipitation, gel filtration on Sephadex G-100 and followed by affinity chromatography on Sepharose 4B, an agglutinin was purified from the hemolymph of T. granosa. The purified lectin contains 5.02% neutral saccharide. The molecular weights is about 123 KDa, which showed 2 bands on SDS-PAGE, and the relative molecular weights of these subunits were 15 KDa and 16 KDa, respectively. In its amino acid composition, Asp has the highest content, followed by Glu and His, but there is a lack of Met. Assays for erythrocyte agglutination and sugar inhibition were done in microtiter plates. The result indicared that TGL-hemolymph could agglutinate erythrocytes from the natural and enzyme-treated erythrocytes of human and animals, and the highest activity was found in the agglutination with rabbit erythrocytes. The agglutinic activity of TGL-hemolymph was inhibited by lactose and D-galactose. The hemagglutination activity of TGL-hemolymph depended on Ca2+, and it showed the highest hemagglutination activity in pH 7.0. Thermostability was not high. The agglutinating activities changed from 2(5) to 2(1) between 30-70 degrees C, and was completely destroyed beyond 80 degrees C. PMID- 17117557 TI - [Detection of formaldehyde induced developmental toxicity assessed with single cell gel electrophoresis]. AB - To understand the developmental toxicity and the maternal toxicity of formaldehyde (FA), pregnant mice were injected celiac with FA, from the 6th day to the 19th day after the pregnancy. After the birth of fetal mice, the single cell gel electrophoresis (SCGE) was applied to test the DNA damage in liver cells of the maternal mice and of the fetal mice. The experimental results obtained from fetal mice indicate that FA does not induce DNA damage at the low concentration (0.2 mg/kg, P > 0.05), while it could induce DNA breakage at higher concentration as 1.0 mg/kg (P < 0.01). Moreover, DPC(DNA-protein crosslinks) was also initiated at the concentration as 2.0 mg/kg (P > 0.05). When the concentration was up to 20.0 mg/kg, almost all of the DNA is completely joined to form DPC (P > 0.05). The experimental results of pregnant mice indicate that DNA breakage could initiate at relatively low concentration (0.2 mg/kg). Meanwhile, the damage effect was obviously enhanced with the increasing of concentrations (1.0 mg/kg, P < 0.01; 2.0 mg/kg, P < 0.01; 20.0 mg/kg, P < 0.01). No significant increase in DPC formation was observed in pregnant mice, indicating that of all the concentrations of FA tested in the experiment, the DNA damage mainly attributes to DNA breakage. These results suggested that FA has both the maternal toxicity and the developmental toxicity. PMID- 17117558 TI - [Functional analysis of rice P0491E01 gene regulating anther development]. AB - The rice P0491E01 gene shares high similarity in amino acid sequence with Arabidopsis gene AtDAD1 (DEFECTIVE IN ANTHER DEHISCENCE1) which plays a key role in the biosynthesis of jasmonic acid. In this paper, we investigated the function of this gene in the anther development of rice using RNA interference strategy. An exon fragment of 263bp was cloned from genomic DNA of P0491E01 to construct RNAi vector pP0491RNAi. Then, pP0491RNAi was transformed into O. sativa japonica by Agrobacterium-mediated transformation and ten transgenic plants were obtained. GUS-staining and PCR analysis confirmed that T-DNA was integrated into rice genome. Three of the transgenic plants were male sterile, and the other transgenic plants showed reduced fertility. Cytological observation indicated that anther development in the early stage of male sterile transgenic plants was normal, however, the microspores could not develop into mature pollen grains. Further investigations of the expression of P0491E01 gene in these transgenic lines by RT-PCR revealed that its transcription was significantly reduced. The results suggest that P0491E01 may play an important role during the late stage of anther development. PMID- 17117559 TI - [Improvement of two-dimensional gel electrophoresis of total proteins from rice anthers]. AB - This paper reported an improvement in 2-D gel electrophoresis of the proteome in Honglian cytoplasmic male sterile rice. An IPGphor unit with immobile pH gradient strips was used as the first dimension and SDS-PAGE as the second. The total anther proteins were extracted using TCA/acetone and then were washed 5-6 times with acetone till the proteins were white and clean, and then tributylphosphine and DTT were added into the rehydration buffer to improve the solubility of the proteins. The 2-D gel was stained by both methods of coomassie blue G-250 and silver. Extraction of proteins, pH of the strips and rehydration of the strips were optimized and compared. Higher repeatability and better separating protein pattern could be gained by this technique. PMID- 17117560 TI - [The study of separating murine embryonic fibroblast from embryonic stem cells by the differential adhesion method]. AB - This study has established a simple method of separating murine embryonic fibroblasts (MEF) from embryonic stem cells (ES cells). The adhesion rates of MEF and ES cells were compared to find out the opitimum time of removing MEF. The result shows that 1.5h and 0.5% gelatin concentration is the opitimum condition of removing MEF. Furthermore, the ES cells which have been purified have the same plating efficiency and the ability of three germ layers differentiation as the unpurified ES cells. After differential adhension, there is strong ALP activity in ES cells. All the results show that the purified ES cells are still in the state of undifferentiation and maintain the pluripotent. PMID- 17117561 TI - [Generation and characterization of the monoclonal antibody and scFv against yeast telomeric guanine-quadruplex DNA]. AB - Most eukaryotic telomeres contain many tandem repeats of G-rich sequence. In budding yeast, Sacchromyces cerevisiae, the G-rich sequence can form parallel stranded quadruplex comformation (G4-DNA) in vitro. Whether this structure exsits in vivo is unknown. To address this question, we generated the antibodies against the G4-DNA of the S. cerevisiae by immunizing the BALB/c mouse with in vitro synthesized G4-DNA oligonucleotides. The antibodies recognize G4-DNA, as well as G-rich DNA sequence in vitro. In order to improve the affinity to G4-DNA substrate, we cloned V(H) and V(L) genes of the antibodies and constructed the single-chain antibody fragment (scFv) with a peptide linker. The recombinated scFv was successfully expressed in E. coli and purified by the affinity chromotography. Based on the sequence of the scFv, we proposed the structure of the antibody by computer-remodeling. The engineered antibodies will be used to detect the existence of the existence of the G4-DNA structure in vivo. PMID- 17117562 TI - [Evaluation of therapeutic effects of synthetic auricular point therapy for treatment of common migraine at the attack stage]. AB - OBJECTIVE: To observe and evaluate the short-term and long-term curative effects of the synthetic auricular point therapy on the common migraine in attack stage. METHODS: With the study method of randomized controlled trial, 108 cases of common migraine at the attack stage were randomly assigned to a treatment group of 72 cases treated with synthetic auricular point therapy (blood-letting on the ear back, point injection of own blood, pricking ear point), and a control group of 36 cases treated with Western medicine. Their short-term and long-term curative effects were observed and evaluated. RESULTS: The total short-term effective rate and the total long-term effective rate were respectively 98.6% and 79. 6% in the treatment group with less adverse effects, and with a very significant difference as compared with the control group (P < 0.01). CONCLUSION: The synthetic auricular point therapy has obvious short-term and long-term curative effects on common migraine at the attack stage with safety. PMID- 17117563 TI - [Effects of electroacupuncture (EA) at early stage on functions of the limbs in the postoperative patient of hypertensive hemorrhage]. AB - OBJECTIVE: To investigate the rehabilitation effect of electroacupuncture (EA) at early stage on hemiplegia in the postoperative patient of hypertensive hemorrhage. METHODS: Seventy-two cases of postoperative patient of hypertensive hemorrhage were divided into an EA group (n = 42) treated with EA at Quchi (LI 11), Shousanli (LI 10), Hegu (LI 4), Zusanli (ST 36), etc., from 1-3 days after operation, twice each day; and a control group (n = 30) with functional exercises. Their rehabilitation effects were compared after treatment of one month. RESULTS: The rehabilitation effect in the EA group was significant better than that in the control group (P < 0.01). CONCLUSION: EA at early stage has good recovery effect for the patient of hemiplegia after operation of hypertensive hemorrhage. PMID- 17117564 TI - [A randomized controlled trial of acupuncture treatment of acute ischemic stroke]. AB - OBJECTIVE: To explore effects of acupuncture on ability of daily living (ALD) and the incidence rate of disability and mortality of the patient of acute ischemic stroke. METHODS: Forty patients with acute ischemic stroke were randomly assigned to an acupuncture group and a control group, 20 cases in each group. The treatment group were treated with acupuncture for 3-4 weeks, 5 times each week, and routine therapy. The control group were treated with routine therapy alone. RESULTS: No statistically significant differences between the two groups in the score of neurological defection, and the incidence rate of disability and mortality at following survey of 3 and 6 months were found. CONCLUSION: Acupuncture is safe and feasible for stroke at early stage. PMID- 17117565 TI - [Effects of a combined regime of auricular-plaster and body acupuncture in treatment of cervical spondylosis of vertebral artery type]. AB - OBJECTIVE: To study effects of a combined regime of auricular-plaster and body acupuncture in treatment of cervical spondylosis of vertebral artery type and make a preliminary revelation of the mechanism. METHODS: Ninety-two patients were randomly divided into 2 groups, the treatment group (n = 56) received the combined regime of auricular-plaster and body acupuncture, and the control group (n = 36) received treatment with body acupuncture. Clinical symptoms and signs, therapeutic effect and some indexes about vertebrobasilar hema-kinetics and hema rheology were investigated before and after treatment. RESULTS: The treatment group was better than the control group in the clinical overall effective rate (89.29%) and the clinically control rate (17. 85%), and in improving the following indexes, including dizziness and headache, the vertebrobasilar volume and rate of blood flow etab and IR (P < 0.05). CONCLUSIONS: A combined regime of auricular-plaster and body acupuncture ameliorates not only main signs but also some indexes about vertebrobasilar hema-kinetics and hema-rheology. This treatment is an effective therapy for cervical spondylosis of vertebral artery type both in Malaysia and in China. PMID- 17117566 TI - [Clinical observation on combination of acupuncture with medication for treatment of spontaneous tremor]. AB - OBJECTIVE: To observe clinical therapeutic effect and safety of combination of acupuncture with medication for treatment of spontaneous tremor. METHODS: With randomized controlled trial method, 26 cases of spontaneous tremor were divided into a treatment group (n = 13) treated with acupuncture plus western medicine, and a control group (n = 13) treated with simple western medicine. They were treated for 6 weeks. Scores of tremor before and after treatment and adverse effects were investigated. RESULTS: The total effective rate of 84.6% in the treatment group was better than 61.5% in the control group (P < 0.05) and there was less adverse effects in the treatment group. CONCLUSION: Combination of acupuncture with medication is a safe and effective therapy for spontaneous tremor. PMID- 17117567 TI - [Clinical observation on photoelectric treatment instrument combined with acupuncture for treatment of simple obesity]. AB - OBJECTIVE: To observe therapeutic effect of photoelectric treatment instrument on simple obesity. METHODS: Sixty cases of simple obesity were randomly divided into two groups, 30 cases in each group. The treatment group were treated with photoelectric treatment instrument combined with acupuncture, i.e. after acupuncture, the electrodes of XS-998A photoelectric treatment instrument were placed at points of the abdomen, with the laser out-put head on Shenque (CV 8), Shuifen (CV 9) or the parts with much fat, or close to the liver area for the patient of fat liver. The control group were treated with routine acupuncture alone. Changes of main symptoms and signs, and BMI before and after treatment were investigated in the two groups. RESULTS: The symptoms and signs, and BMI were more significantly improved in the treatment as compared with the control group with a significant difference between the two groups (P < 0.05). CONCLUSION: Acupuncture combined with laser radiation at the relative area on the abdomen can increase the therapeutic effect on obesity. PMID- 17117568 TI - [Effects of Ziwuliuzhu combined selection of the source point and the collateral point on athletic injuries and the state of channels]. AB - OBJECTIVE: To probe into therapeutic effects of Ziwuliuzhu combined selection of the source point and the collateral point on athletic injuries and the state of channels. METHODS: Ninety cases meeting diagnostic criteria were randomly divided into a group of Ziwuliuzhu combined selection of the source point and the collateral point (group A), a group of routine selection of acupoint (group B) and a group of external application of medicine (group C), 30 cases in each group. The electric conduction amount on source points of 12 channels were determined before and after treatment with a point diagnosis and treatment instrument. Changes of the state of channels and clinical therapeutic effects in the 3 groups were investigated after treatment. RESULTS: The therapeutic effect in the group A was better than those in the other two groups (P < 0.05). CONCLUSION: Ziwuliuzhu combined selection of the source point and the collateral point has good effect on pain and other clinical symptoms of athletic injuries, and makes channels of imbalance tend to balance or recover balance. PMID- 17117569 TI - [Clinical observation on auricular point sticking for treatment of 25 children of semiluxation of circo-axis vertebrae]. AB - OBJECTIVE: To search for the best method for child semiluxation of circo-axis vertebrae. METHODS: Fifty cases were randomly divided into a treatment group of 25 cases treated with auricular point sticking, and a control group of 25 cases treated with normal acupuncture. RESULTS: The total effective rate was 92.0% in the treatment group and 64.0% in the control group with a significant difference between the two groups (P < 0.05). CONCLUSION: Auricular point sticking has a good effect on child semiluxation of circo-axis vertebrae. PMID- 17117570 TI - [Clinical observation on treatment of chloasma with point embedding thread]. AB - OBJECTIVE: To observe therapeutic effect of point embedding thread on chloasma. METHODS: Thirty cases of the treatment group were treated with embedding thread at body points for 6 times, points of the face for once, pricking blood therapy on ear points once each week; the 30 cases of the control group with body acupuncture and ear acupuncture, twice each week. After treatment of 4 months, their therapeutic effects on chloasma and companied symptoms were evaluated. RESULTS: The effective rate on chloasma was 100.0% in the treatment group and 70.0% in the control group with a very significant difference between the two groups (P < 0.01). The effective rates on the companied symptoms were 100.0% in the two groups, but the cured and markedly effective rate in the treatment group was better than that in the control group (P < 0.01). CONCLUSION: The therapeutic effect of point embedding thread combined with ear point pricking blood therapy on chloasma and its companied symptoms is better than that of the body acupuncture combined with ear acupuncture. PMID- 17117571 TI - [Acupuncture combined with massage for treatment of irritable bowel syndrome]. AB - OBJECTIVE: To observe the therapeutic effect of acupuncture combined with massage on irritable bowel syndrome. METHODS: Ninety-three cases of irritable bowel syndrome were randomly divided into three groups: group of acupuncture combined with massage (group A), simple acupuncture control group (group B), simple massage control group (group C), 31 cases in each group. Their therapeutic effects were compared. RESULTS: The cured rate and the effective rate in the group A were significantly higher than those in both group B and group C (P < 0.05). CONCLUSION: Acupuncture combined with massage therapy shows a better therapeutic effect on irritable bowel syndrome, and they should be jointly used in clinical treatment. PMID- 17117572 TI - [Clinical study on round-sharp needle therapy combined with massage for prolapse of lumbar intervertebral disk]. AB - OBJECTIVE: To search for a better therapy for prolapse of lumbar intervertebral disk. METHODS: One hundred and sixteen cases were randomly divided into a treatment group treated with round sharp needle plus massage, and a control group with filiform needle plus massage, 58 cases in each group. Changes of symptoms and signs were observed before and after treatment. RESULTS: The effective rate was 98.2% in the treatment group and 82.8% in the control group (P < 0.01). CONCLUSION: Round sharp needle combined with massage has a better therapeutic effect on prolapse of lumbar intervertebral disk. PMID- 17117573 TI - [Effect of acupuncture and massage at tendon on F-wave of electromyogram in the patient of flexor spasm of the upper limb after operation of cervical spondylosis]. AB - OBJECTIVE: To explore the mechanism of acupuncture and massage at the tendon in treatment of the upper limb flexor spasm after cervical operation. METHODS: Fifty six cases were randomly divided into a treatment group (n = 30) treated with acupuncture and massage at the tendons, and a control group treated with traditional acupuncture and medication. Changes of F-wave, time limit and threshold value in electromyogram before and after treatment were investigated in the both groups. RESULTS: Before treatment, the F-wave in the patient of the upper limb spasm post-cervical operation was characterized by high amplitude, long time limit and low threshold value, with no significant difference between the two groups (P > 0.05); after treatment, the amplitude lowered (P < 0.01), the time limit shortened (P < 0.001) in the treatment group and no significant changes were found in the control group (P > 0.05), but the amplitude of F-wave in the treatment group was significantly lower than that in the control group (P < 0.05). The threshold value significantly increased in the two groups, with the treatment group more significantly increased than the control group (P < 0.05). CONCLUSION: Acupuncture and massage at tendons can relieve the spasm through lowing the excitement in the cells of the anterior horn of the spinal cord, decreasing the amplitude and time limit of the F-wave, increasing the threshold value and reducing traction reflection. PMID- 17117574 TI - [Observation on therapeutic effects of different needling methods at tiaokou (ST 38) on periarthritis of shoulder]. AB - OBJECTIVE: To compare therapeutic effects of different needling methods at Tiaokou (ST 38) on periarthritis of shoulder. METHODS: Eighty cases of periarphritis of shoulder were randomly assigned to four groups. Tiaokou (ST 38) was selected in all the 4 groups. The treatment group were treated with penetration needling, and the control group A, B, C were treated by needling into 40 mm deep, needling into the subcutaneous tissue, sham acupuncture, respectively. Therapeutic effect was evaluated by Constant-Murley shoulder joint scale. RESULTS: The clinical effective rate was 100.0% in the treatment group and 68.4% in the control group A, 52.6% in the control group B, 38.0% in the control group C with significant differences as the treatment group compared with the control groups A, B, C (P < 0.05 or P < 0.01). And with no significant difference as the control group A compared with the control group B, C, and the group B with the group C (P > 0.05). CONCLUSION: Different needling depths of Tiaokou (ST 38) have different therapeutic effects, the penetration needling being the best and the sham needling being worse which can be used as control method. PMID- 17117575 TI - [Effect of acupuncture on pH values of acupoints in the goat]. AB - OBJECTIVE: To study effect of acupuncture on pH values of acupoints and non acupoints. METHODS: The pH values of acupoints and non-acupoints were determined with pH sense needles at the same time and separately in the goat. RESULTS: (1) The pH value of the acupoints and non-acupoints decreased when acupuncture was separately at acupoints and non-acupoints with the pH value of the acupoint decreased more significantly; (2) there was no significant difference in decreasing amplitudes of the pH values between the acupoint and non-acupoint when acupuncture was simultaneously given at acupoints and non-acupoints. CONCLUSION: Acupuncture can more significantly strengthen metabolism of acupoints; when acupuncture was simultaneously given at acupoints and non-acupoints, local metabolism is influenced each other. PMID- 17117576 TI - [Effects of eight methods of intelligent turtle on free radical metabolism in the kidney-yang deficiency model guinea pigs]. AB - OBJECTIVE: To observe effects of moxibustion according to opening points on time in eight methods of intelligent turtle on aging and the mechanism. METHODS: Healthy and adult guinea pigs were randomly divided into 5 groups, a prevention group of kidney-yang deficiency model with eight method of intelligent turtle (group I), a prevention group of kidney-yang deficiency model (group II) with moxibustion at Mingmen (GV 4), a group of treatment group of kidney-yang deficiency model with eight methods of intelligent turtle (group III), a treatment group of kidney-yang deficiency model (group IV) with moxibustion at Mingmen (GV 4) and a control group of kidney-yang deficiency. Effects of moxibustion according to opening on time in eight methods of intelligent turtle and moxibustion at Mingmen (GV 4) on superoxide dismulase (SOD) activity and malondialdehyde (MDA) level were compared. RESULTS: After modeling, no signs of kidney-yang deficiency and no significant changes of SOD and MDA were found in group I and II; but obvious symptoms of kidney-yang deficiency and decrease of SOD (P < 0.01) and increase of MDA (P < 0.01 or P < 0.05) were found in group III, IV and V; after treatment, in the group III); the symptoms of kidney-yang deficiency basically disappeared, SOD increased (P < 0.01) and MDA decreased (P < 0.01), with no significant differences as compared with those before modeling (P > 0.05); in the group N, the symptoms of kidney-yang deficiency significantly improved and SOD increased (P < 0.05) with difference as compared with those before modeling (P < 0.05), and MDA decreased (P < 0.05), but with no significant difference compared with that before modeling (P > 0.05); the symptoms of kidney yang deficiency, SOD and MDA did not significantly change in group V (P > 0. 05). CONCLUSION: Both moxibustion according to opening points on time in eight methods of intelligent turtle and moxibustion at Mingmen (GV 4) can effectively regulate free radical metabolism and have obvious action of preventing and treating kidney yang deficiency in the model guinea pigs, and the therapeutic action of moxibustion according to opening points on time in eight methods of intelligent turtle is better than that of moxibustion at Mingmen (GV 4). PMID- 17117577 TI - [Simple analysis on professor ZHANG Dao-Zong's academic thought of dredging the Governor Vessel and regulating mentality]. AB - Introduce Professor Zhang Dao-zong's studies on the Governor Vessel and its academic thought of dredging the Governor Vessel and regulating mentality, and clinical experiences of dredging the Governor Vessel and regulating mentality therapy for apoplexy, epilepsy, vertigo, ankynosing spondylitis, traumatic paraplegia, child Tourette's disease, etc.... PMID- 17117578 TI - [Professor MA Ji-Xing's contribution in the field of acupuncture and moxibustion science]. AB - Prof. MA, a famous medical historian and a founder of TCM literature science in our country. He works industriously and in the past over 60 years, he has attained great achievements in the field of acupuncture and moxibustion, and made great contribution to the acupoint science and channel and collateral science, published The Concise Acupuncture and Moxibustion Bone-setting, color teaching hanging chart of The Reference Chart of Anatomic Positions of Stimulating Points for Acupuncture and Moxibustion Therapy, Bronze Acupuncture Figure and Point Location of the Bronze Figure, Textual Criticism of Ancient Medical Books in Dunhuang, Studies on Unearthed Ancient Medical Books No Longer Extant, etc., which have important influences on studies of acupuncture and moxibustion history and acupuncture and moxibustion literature. PMID- 17117579 TI - [Present situation and thinking of studies on apoplectic aphasia]. AB - There were different emphasis between Chinese medicine and Western medicine in the study of apoplectic aphasia, the study of Chinese medicine lay particular emphasis on treatment with more interfering methods but lacking unifying evaluation criteria; while studies of Western medicine lay particular emphasis on studies of mechanisms, classification and language rehabilitation of aphasia, etc.. Selection and application of scientific study methods are key to attain expectation results of studies, and deeply make studies of apoplectic aphasia, so as to explore and establish the model of integrated Chinese and western medicine study of aphasia, promote rehabilitation of language function and increase life quality of the patient of apoplexy. PMID- 17117580 TI - [Preliminary study on characteristics of the Illustrations of Moxibustion in Dunhuang Caves in treatment of 5 kinds of strain and 7 kinds of impairments]. AB - The Illustrations of Moxibustion, a remained book in Dunhuang Caves, is available the earliest monograph about moxibustion methods with both pictures and writing since recorded writing. In treatment of 5 kinds of strain and 7 kinds of impairments, point association for correspondence between man and universe was adopted human point distribution according to Jiugong and Bagua; and point association method of viewing the situation as a whole according to humam point distribution on the head, the body and the limbs; it wrote and drew available the earliest naked figures for acupuncture and moxibustion in simple natural materialism; and it is the most reasonable, the most complete explanation of sexual science in Chinese medicine, and is a sample for treatment of 5 kinds of strain and 7 kinds of impairments by ancient physicians stressing moxibustion, with convenient point selection and safety. PMID- 17117581 TI - [Advances of modern studies of acupuncture and moxibustion for treatment of ovulation disorders]. AB - OBJECTIVE: To introduce the modern advances of studies on acupuncture and moxibustion for treatment of ovulation disorders. METHODS: The recent years' literature of acupuncture and moxibustion for treatment of ovulation disorders were reviewed from clinical studies and treatment mechanisms. RESULTS: Acupuncture and moxibustion can regulate the hypothalamus-pituitary-gonadal axis, the nervous system and immune system of the organism. CONCLUSION: Acupuncture and moxibustion has very good clinical effects on ovulation disorders, and it is a safe and effective adjuvant way for modern assisted reproductive technique. PMID- 17117582 TI - The human genome project: understanding the role of inflammation in disease and disease prevention. PMID- 17117583 TI - Histone deacetylase inhibitors as novel anti-inflammatory agents. AB - Histone acetylation regulates inflammatory gene expression and also plays a role in diverse functions such as DNA repair and cell proliferation. Changes in histone acetylation patterns have been reported in many human diseases, particularly cancer, and investigators have used histone deacetylase inhibitors (eg, suberoylanilide hydroxamic acid and MS-275) against many malignancies. In vitro and in vivo data in a number of cell types and animal models of disease indicate that selective histone deacetylase inhibitors may have the potential to act as anti-inflammatory agents possibly acting through non-histone proteins. More evidence is required concerning the long-term safety of these agents, for example, lung destruction, and in comparison with currently used therapies. PMID- 17117584 TI - Marine sponge metabolites for the control of inflammatory diseases. AB - Marine organisms are a rich source of bioactive metabolites. A number of potential anti-inflammatory compounds have been isolated from marine invertebrates that exhibit phospholipase A2 inhibitory activity. A wide range of marine compounds have been investigated for their anti-inflammatory properties. Cacospongionolide B and petrosaspongiolide M are representative examples of anti inflammatory compounds in experimental models of acute or chronic inflammation. The mechanisms of action of these compounds include phospholipase A, inhibition as well as the control of nuclear factor-B activation and inflammatory gene expression. Although many marine compounds exhibit interesting anti-inflammatory properties, few have entered clinical trials. The future development of this class of compounds as anti-inflammatory drugs requires the introduction of novel molecular targets of therapeutic relevance in addition to biotechnological approaches for the production of these molecules. PMID- 17117585 TI - Imatinib mesylate for the treatment of hypereosinophilic syndromes. AB - Hypereosinophilic syndromes (HES) define several distinct entities, all of which show blood and tissue hypereosinophilia and end-organ secondary manifestations. From a clinical and pathogenic perspective, the best characterized are the myeloproliferative and lymphoproliferative variants. While there are several therapies available for the treatment of HES, such as corticosteroids, IFNalpha and cytotoxic agents, there remains a high incidence of side effects and inconsistent efficacy. Abnormal tyrosine kinase resulting from the FIP1L1PDGFRA fusion gene is found in many patients with myeloproliferative variant, and is currently best targeted therapeutically with imatinib mesylate, a tyrosine kinase inhibitor. However, complete hematological remissions obtained with this compound in patients without this defect suggest that it could be effective in other subtypes of HES. PMID- 17117586 TI - RAGE in inflammation: a new therapeutic target? AB - High-molecular group box 1-protein, S100/calgranulins, advanced glycation end products (AGEs), amyloid-beta peptides and the family of beta-sheet fibrils contribute to a number of inflammatory conditions by promoting cellular dysfunction and breaking immune tolerance. The receptor of AGE (RAGE) is a multiligand receptor of the immunoglobulin superfamily of cell surface molecules that acts as a pattern recognition receptor. Besides binding ligands actively participating in inflammation and immune responses, RAGE serves as an endothelial adhesion receptor for leukocyte integrins and promotes leukocyte recruitment and extravasation of infiltrating cells. Engagement of RAGE subsequently converts transient cellular stimulation into sustained cellular dysfunction driven by long term activation of the proinflammatory nuclear factor-kappaB. Deletion of RAGE and pharmacological interventions targeting interruption of RAGE-ligand interaction suppresses inflammation and dampens tissue damage in experimental models of inflammatory disorders, thus delineating RAGE as a potential therapeutic target in inflammation. PMID- 17117587 TI - Current concepts and treatment options in eosinophilic esophagitis. AB - Eosinophilic esophagitis (EE) is an inflammatory disorder of the esophagus that affects both children and adults, and is different from gastro-esophageal reflux disease. The immunopathogenesis of EE involves an allergic response to environmental and food allergens, and the proinflammatory cytokines IL-5 and IL 13. EE may be associated with atopic disorders and peripheral eosinophilia, and may be familial in distribution. The most common presentation is dysphagia and food impaction in adults, with additional manifestations of epigastric pain, emesis, weight loss and failure to thrive in children. Typical endoscopic findings include ringed esophagus, linear furrows, strictures and narrow esophagus. Diagnosis is confirmed by the presence of 20 or more eosinophils per high power field in the esophagus. The available treatment options include elemental diet, avoidance of specific food allergens, topical and systemic corticosteroids, and humanized monoclonal antibodies against IL-5. This review summarizes the etiopathogenesis, clinical, endoscopic and histopathological findings in EE, and describes current available treatment options. PMID- 17117588 TI - Estrogen receptor ligands in the control of pathogenic inflammation. AB - Inflammation is recognized as a key component in a number of diseases, including rheumatoid arthritis, inflammatory bowel disease and atherosclerosis. Although well known for their classic effects on the reproductive tract and action by means of estrogen response elements in gene promoters, estrogens are also known to possess anti-inflammatory activity. This was originally highlighted with the observation that pregnancy ameliorates symptoms of rheumatoid arthritis, multiple sclerosis and inflammatory bowel disease. Furthermore, the antagonistic cross talk between nuclear factor kappaB and estrogen receptor signaling pathways has been well documented. Recently, novel estrogen receptor ligands, pathway selective ligands and estrogen receptor beta-selective ligands have been identified which demonstrate potent anti-inflammatory activity; these ligands are being analyzed for their therapeutic potential in pathogenic inflammation. PMID- 17117589 TI - Dissecting the mechanism of T-cell anergy with immunophilin ligands. AB - T-cell receptor engagement in the absence of costimulation leads to T-cell anergy. The biochemical and molecular mechanisms responsible for this form of T cell tolerance are becoming better defined. This review examines the intersection between T-cell receptor-induced anergy and the immunophilin ligands ciclosporin A, FK-506, rapamycin and sanglifehrin A, and focuses on how these compounds play an important role in dissecting the pathways leading to the induction and maintenance of anergy. Finally, the clinical role of these compounds as immunosuppressive agents will be discussed in the context of their effects on promoting or inhibiting T-cell anergy. PMID- 17117590 TI - The immunological synapse as a novel therapeutic target. AB - The onset of adaptive immune responses includes the presentation of foreign antigenic peptides to T-cells, and the formation of a T-cell-antigen-presenting cell interface termed the immunological synapse (IS). Although the generation of a mature IS is thought to be the hallmark of T-cell activation, new evidence suggests that microclusters ofat signaling molecules at the periphery of the IS are responsible for initiating and maintaining T-cell activation while the core of the IS provides a platform for signal downregulation. In this context, costimulatory molecules and self-peptides contribute to sustain the signaling required for T-lymphocyte differentiation into effector cells. This review discusses these aspects in the identification of novel candidates for therapeutic modulation of immune responses. PMID- 17117591 TI - Drug evaluation: apratastat, a novel TACE/MMP inhibitor for rheumatoid arthritis. AB - Wyeth Research was developing apratastat (TMI-005), one in a series of dual TNFalpha-converting enzyme and matrix metalloprotease-13 inhibitors, for the potential treatment of inflammation, especially rheumatoid arthritis. By January 2005, apratastat had entered a phase II clinical trial; however, in October 2006, Wyeth reported that it had terminated development of the drug because of lack of efficacy in this trial. PMID- 17117592 TI - Drug evaluation: VX-702, a MAP kinase inhibitor for rheumatoid arthritis and acute coronary syndrome. AB - Vertex Pharmaceuticals Inc, in collaboration with Kissei Pharmaceutical Co Ltd, is developing VX-702, one of a series of second-generation, orally active p38 MAP kinase inhibitors, for the potential treatment of inflammation, rheumatoid arthritis and cardiovascular diseases. In June 2005, a phase II clinical trial of VX-702 was initiated in rheumatoid arthritis. In July 2006, Vertex was planning to file an IND in the second half of 2006. PMID- 17117593 TI - A standard of knowledge for pharmaceutical sales representatives. PMID- 17117594 TI - The critical path initiative: what is in it for P&T Committees? PMID- 17117595 TI - Gender discrepancies in the level of disease progression in hospitalized patients with diabetes. AB - Diabetes is a widely recognized and common cause of mortality, morbidity, and excess cost in the health care system. Type 2 diabetes is specifically on the rise, particularly in association with the increasing obesity rate. Substantial literature has examined differences in behavior across genders and potential effect on disease. However, based on a literature review, little has been done to evaluate differences in patterns of diabetes related to inpatient care across genders and to link these differences with potential explanations related to differences in care-seeking behavior, treatment compliance, and lifestyle choices. This review and analysis of national inpatient data demonstrates substantial differences associated with gender in patterns of hospital-based care in patients with type 2 diabetes. Men are more likely to be admitted for diabetes related conditions and to present with blood sugars at extreme levels, including acutely elevated and dangerous blood sugar levels. When considering these findings in conjunction with the existing literature, it can be concluded that these differences are primarily reflective of variances in care-seeking behavior and long-term adherence to prescribed medications. Policies that promote expansion of health care benefits to include coverage for men at risk for type 2 diabetes, aggressive education and treatment programs, and expanded prescription drug coverage are necessary to reduce gender discrepancies in patterns of hospitalization and to improve outcomes. PMID- 17117596 TI - New recommendations from the Institute of Medicine on preventing medication errors. PMID- 17117597 TI - Managed care and clinical autonomy in the workers' compensation market. AB - Despite increases in health care premiums, the effect of relaxing cost containment mechanisms on health care utilization is not yet well understood at the microlevel. This study used a regulatory change in the California workers' compensation system to examine the effect of relaxing broad-based utilization management constraints and increasing clinical autonomy on methods of treatment and service intensity, and compared the responses of managed care network and fee for-service providers. Between 1993 and 2000, the likelihood of a fee-for-service claim receiving a chiropractic treatment increased from 22% to 32%, the likelihood of receiving diagnostic radiology decreased from 24% to 15%, and the likelihood of receiving physical medicine with diagnostic services remained relatively stable. Treating fee-for-service claims with network care would have decreased the likelihood of receiving manipulations by 13 percentage points and physical medicine with diagnostic services by two percentage points. The likelihood of receiving office-visit-only treatment would have increased by 130% (14 percentage points), and the likelihood of receiving a diagnostic radiology treatment would have increased by 28% (4 percentage points). Treatment by network providers would have reduced the number of office visits by 18%, diagnostic radiology and ultrasound exams by 26%, passive physical medicine procedures by 40%, active physical medicine procedures by 43%, physical medicine assessments by 45%, and chiropractic treatments by 46%. PMID- 17117598 TI - A two-stage decision analysis to assess the cost of 5-aminosalicylic acid failure and the economics of balsalazide versus mesalamine in the treatment of ulcerative colitis. AB - Ulcerative colitis (UC) is a costly disease, especially if not properly treated. Epidemiologic studies have shown that many patients progress in one year from initial treatment with prednisone to expensive colonectomy surgery if the UC is not managed with drug therapy. A two-stage decision analysis was conducted to (1) estimate the cost of a 5-aminosalicylic acid (5-ASA) treatment failure using the treatment guidelines recommended by the American College of Gastroenterology and (2) incorporate the cost of a 5-ASA treatment failure to determine which oral 5 ASA agent results in cost minimization and cost effectiveness. The analysis was conducted from the payer perspective, incorporating results from clinical trials directly comparing oral balsalazide capsules and a specific formulation of oral mesalamine. Health care costs related to UC for an oral 5-ASA failure is greater than dollar 11,500 on average in the first six months after therapy. Patients treated with balsalazide capsules had 16% lower total direct health care costs, 32% better outcomes (days without symptoms or steroids), and 37% greater cost effectiveness compared with patients treated with a specific formulation of oral mesalamine. Coordinated efforts should be taken to avoid the cost and morbidity associated with 5-ASA treatment failures. PMID- 17117599 TI - Focus on methods: chi-square analysis. PMID- 17117600 TI - Allocation of fault required in 'comparative negligence' cases. Haley v. Brown, No. 93, 148 (Kan. App. 08/25/2006) P.2d -KS. PMID- 17117601 TI - RN not at fault for failure to give epinephrin. Taylor v. Jackson-Madison Cnty. Gen. Hosp., No.W2005-02471-COA-R3-CV (Tenn. App. 8/23/06) -Tn. PMID- 17117602 TI - LA: alcohol treatment pt. fled, fell, and died: RN calling dr. re patient's vitals not liable. Ball V. Behavioral, no. 41,329-CA (LA.App.Cir.2 08/23/06) So.2d -LA. PMID- 17117603 TI - Should nurses have questioned dr.'s orders? Cerny V. Williams, No. 2004-10053 (09/19/2006) -NY. PMID- 17117604 TI - Contribution of immunohistochemistry in prognostic assessment of epithelial ovarian carcinoma --review of the literature I. AB - Epithelial ovarian carcinoma is worldwide the sixth most common female cancer, and this malignancy carries the highest mortality among all gynecological cancers. The high mortality is due mostly to the fact that the tumor is frequently diagnosed late, in advanced stage, as the early disease is often asymptomatic and no effective screening methods are available. The most important prognostic factors in ovarian carcinoma are the stage, size of residual tumor following surgery, presence of ascites, age and the general condition of the patient, tumor histology, and, in patients with early disease, also the grade of the tumor. Large number of studies on prognostic and predictive factors in epithelial ovarian carcinoma has been published, often with contradictory results. The most intensely studied prognostic factors are those for expression of hormonal receptors, for tumor proliferation activity (mainly by antigen Ki-67 and topoisomerase IIalpha), the markers of apoptosis (p53, p21, mdm2, bcl-2 and other proteins), or other oncoproteins (particularly HER-2/neu). PMID- 17117605 TI - IMRT with the use of simultaneous integrated boost in treatment of head and neck cancer: acute toxicity evaluation. AB - Acute toxicity has been evaluated in head and neck cancer patients treated with intensity-modulated radiotherapy using simultaneous integrated boost (SIB-IMRT). The basis of the treatment protocol is an irradiation in 30 fractions with a total dose: 66 Gy to the region of macroscopic tumor, 60 Gy to the region of high risk subclinical disease and 54 Gy to the region of low-risk subclinical disease. Between December 2003 and September 2005, 38 patients with carcinoma of different locations in the head and neck region were irradiated. Five patients underwent concurrent chemotherapy (weekly cisplatin). Acute toxicity was evaluated according to Radiation Therapy Oncology Group toxicity scale for skin, mucous membrane, salivary glands, pharynx and esophagus and larynx. All 38 patients completed the therapy without urgency of interruption due to acute toxicity of radiotherapy. No patient experienced grade 4 toxicity. More severe toxicity was observed in patients with concurrent chemotherapy. The results confirm that the irradiation according to our SIB-IMRT protocol is a therapy with acceptable toxicity and there is a space for radiobiological enhancement of this regimen by concurrent chemotherapy, e.g. weekly cisplatin. PMID- 17117606 TI - Possibilities of combined surgical treatment of lung tumours and heart diseases. AB - The purpose of the study was a retrospective evaluation of the outcome of surgical therapy of lung cancer in patients where there was concomitant cardiac disease and who underwent a cardiac operation either because of ischemic heart disease or because of valvular disease. These patients were operated on at various time intervals (two to ten months) after their cardiac operation. Some patients had their lung cancer surgery after the cardiac operation because of the high risk of possible cardiac postoperative complications; in one patient the lung operation preceded the cardiac one. PMID- 17117607 TI - Predictors of mortality in patients with acute renal failure. AB - Mortality associated with acute renal failure (ARF) remains high despite of developments in therapy strategies and definition of different prognostic factors. Therefore, this study focused on to define new prognostic factors and especially regional characteristics of the ARF patients. One hundred fifteen ARF patients, diagnosed from November 1998 to May 2003, were included to this prospective and observational study. Clinical features, laboratory parameters, Acute Physiology and Chronic Health Evaluation (APACHE) III scores and co-morbid conditions of the patients were examined. Clinical and laboratory data, and APACHE III scores were recorded at the first nephrology consult day. Thirty of the patients (26%) died. APACHE III scores, presence and the total number of co morbid conditions and serum albumin levels at the time of first nephrology consultation were found as independent predictors of mortality. There was a negative correlation between APACHE III scores and serum albumin levels. Not only increased APACHE III score and presence of co-morbid conditions but also low serum albumin level was found as the predictors of mortality. However, only serum albumin level is seen as modifiable prognostic factor among these parameters. Therefore, further studies are necessary to determine the causes of hypoalbuminemia in patients with ARF and the effect of it's effective treatment on patients outcome. PMID- 17117608 TI - Docetaxel labeled with iodine 131 radionuclide--a possible new anticancer drug? (The pilot study). AB - The cytotoxic potentials of the lipiodol emulsion with dissolved 131I-docetaxel, the 131I-lipiodol emulsion and non-labeled docetaxel were tested on the HeLa Hep2 cell line during 24 hours. The pilot study confirmed that the radio-labeled docetaxel was significantly more toxic than the radionuclide or docetaxel alone. PMID- 17117609 TI - Successful treatment of iron overload with phlebotomies in two siblings with congenital dyserythropoietic anemia--type II (CDA-II). AB - Successful treatment of iron overload by phlebotomies has been reported in two splenectomized siblings with congenital dyserythropoietic anemia--type II (CDA II). In both patients 400 ml of blood were withdrawn every month. During three years 12 200 ml of blood were removed. The serum ferritin levels decreased from 1450,4 microg/L and 1131,7 microg/L to 447 microg/L and 457 microg/l, respectively. The transferrin saturation dropped from 0,99 at the start of the therapy to 0,64 and 0,86, respectively. The values of Hb, Hct, erythrocyte counts and MCV did not change as well as did not change reticulocyte counts, reticulocyte index, and RDW. Both patients tolerated repeated phlebotomies well. The decrease of bilirubin and normal values of haptoglobin might be the concequence of diminished destruction of erythrocytes and their precursors. Our observation confirms that phlebotomies can be used with success in CDA patients with mild anemia as treatment modality of iron overload. PMID- 17117610 TI - Expression profiling of drug response--from genes to pathways. AB - Understanding individual response to a drug-what determines its efficacy and tolerability-is the major bottleneck in current drug development and clinical trials. Intracellular response and metabolism, for example through cytochrome P 450 enzymes, may either enhance or decrease the effect of different drugs, dependent on the genetic variant. Microarrays offer the potential to screen the genetic composition of the individual patient. However, experiments are "noisy" and must be accompanied by solid and robust data analysis. Furthermore, recent research aims at the combination of high-throughput data with methods of mathematical modeling, enabling problem-oriented assistance in the drug discovery process. This article will discuss state-of-the-art DNA array technology platforms and the basic elements of data analysis and bioinformatics research in drug discovery. Enhancing single-gene analysis, we will present a new method for interpreting gene expression changes in the context of entire pathways. Furthermore, we will introduce the concept of systems biology as a new paradigm for drug development and highlight our recent research-the development of a modeling and simulation platform for biomedical applications. We discuss the potentials of systems biology for modeling the drug response of the individual patient. PMID- 17117611 TI - New directions for drug discovery. AB - Modern drug discovery demands an integrative approach, using many different technologies, but ultimately based on an understanding of the pathophysiology of the disease state to be treated. Targeting drugs at the main pathophysiological process is the key to success. This issue needs to be addressed with the multiple screening systems available, which can be used to find new leads. PMID- 17117612 TI - Contributions of molecular biology to antipsychotic drug discovery: promises fulfilled or unfulfilled? AB - This review summarizes the various conceptual paradigms for treating schizophrenia, and indicates how molecular biology and drug discovery technologies can accelerate the development of new medications. As yet, there is no convincing data that a crucial druggable molecular target exists which, if targeted, would yield medications with efficacies greater than any currently available. It is suggested, instead, that drugs which interact with a multiplicity of molecular targets are likely to show greater efficacy in treating the core symptoms of schizophrenia. PMID- 17117613 TI - Membrane transporter proteins: a challenge for CNS drug development. AB - Drug transporters are membrane proteins present in various tissues such as the lymphocytes, intestine, liver, kidney, testis, placenta, and central nervous system. These transporters play a significant role in drug absorption and distribution to organic systems, particularly if the organs are protected by blood-organ barriers, such as the blood-brain barrier or the maternal-fetal barrier. In contrast to neurotransmitters and receptor-coupled transporters or other modes of interneuronal transmission, drug transporters are not directly involved in specific neuronal functions, but provide global protection to the central nervous system. The lack of capillary fenestration, the low pinocytic activity and the tight junctions between brain capillary and choroid plexus endothelial cells represent further gatekeepers limiting the entrance of endogenous and exogenous compounds into the central nervous system. Drug transport is a result of the concerted action of efflux and influx pumps (transporters) located both in the basolateral and apical membranes of brain capillary and choroid plexus endothelial cells. By regulating efflux and influx of endogenous or exogenous substances, the blood-brain barrier and, to a lesser extent the blood-cerebrospinal barrier in the ventricles, represents the main interface between the central nervous system and the blood, i.e., the rest of the body. As drug distribution to organs is dependent on the affinity of a substrate for a specific transport system, membrane transporter proteins are increasingly recognized as a key determinant of drug disposition. Many drug transporters are members of the adenosine triphosphate (ATP)-binding cassette (ABC) transporter superfamily or the solute-linked carrier (SLC) class. The multidrug resistance protein MDR1 (ABCB1), also called P-glycoprotein, the multidrug resistance associated proteins MRP1 (ABCC1) and MRP2 (ABCC2), and the breast cancer resistance protein BCRP (ABCG2) are ATP-dependent efflux transporters expressed in the blood-brain barrier They belong to the superfamily of ABC transporters, which export drugs from the intracellular to the extracellular milieu. Members of the SLC class of solute carriers include, for example, organic ion transporting peptides, organic cation transporters, and organic ion transporters. They are ATP independent polypeptides principally expressed at the basolateral membrane of brain capillary and choroid plexus endothelial cells that also mediate drug transport through central nervous system barriers. PMID- 17117614 TI - Experimental animal models for the simulation of depression and anxiety. AB - An impressive number of animal models to assess depression and anxiety are available today. However, the relationship between these models and the clinical syndromes of depression and anxiety is not always clear. Since human anxiety disorders represent a multifactorial phenomenon frequently comorbid with major depression and/or other psychiatric problems, the chance of creating animal models which consistently reflect the human situation is quite poor. When using experimental models to understand homologies between animal and human behavior, we have to consider the context in which an animal is investigated, and both the functional significance and relevance of the behavioral parameters that are quantified. Moreover, gender and interindividual and interspecies variabilities in behavioral responses to the test situation and in the sensitivity to pharmacological treatments are potential sources for confounding results. In the past, these aspects have been often neglected in preclinical approaches to behavioral pharmacology and psychopharmacology. A pragmatic approach of combined preclinical and clinical efforts is necessary to imitate one or more aspects relevant to pathological anxiety disorders and depression. The resulting models may identify central nervous processes regulating defined behavioral output, with the potential to develop more effective treatments. PMID- 17117615 TI - The role of serendipity in drug discovery. AB - Serendipity is one of the many factors that may contribute to drug discovery. It has played a role in the discovery of prototype psychotropic drugs that led to modern pharmacological treatment in psychiatry. It has also played a role in the discovery of several drugs that have had an impact on the development of psychiatry. "Serendipity" in drug discovery implies the finding of one thing while looking for something else. This was the case in six of the twelve serendipitous discoveries reviewed in this paper, i.e., aniline purple, penicillin, lysergic acid diethylamide, meprobamate, chlorpromazine, and imipramine. In the case of three drugs, i.e., potassium bromide, chloral hydrate, and lithium, the discovery was serendipitous because an utterly false rationale led to correct empirical results; and in case of two others, i.e., iproniazid and sildenafil, because valuable indications were found for these drugs which were not initially those sought The discovery of one of the twelve drugs, chlordiazepoxide, was sheer luck. PMID- 17117616 TI - Surrogate outcomes in neurology, psychiatry, and psychopharmacology. AB - A surrogate outcome can be defined as an outcome that can be observed sooner, at lower cost, or less invasively than the true outcome, and that enables valid inferences about the effect of intervention on the true outcome. There is increasing interest in the use of surrogate outcomes of treatment efficacy measurement in investigational drug trials. However, the significance of surrogate markers of treatment outcome in neurology and psychiatry has not yet been sufficiently demonstrated. Few such markers have been adequately "validated, " that is, shown to predict the effect of the treatment on the clinical outcome of interest. In this article, evidence that would support the validation of such markers is discussed. Biomarkers used during early clinical development programs of new psychotropic compounds are considered in the contexts of Parkinson's disease, affective disorder, and schizophrenia. The particular case of neuroprotective trials is exemplified by Parkinson's disease, where a biomarker substituting for a clinical measure of progression could be considered as a surrogate treatment outcome. PMID- 17117617 TI - Functional genomics in postmortem human brain: abnormalities in a DISC1 molecular pathway in schizophrenia. AB - The disrupted in schizophrenia 1 (DISC1) gene has been identified as a schizophrenia susceptibility gene based on linkage and single nucleotide polymorphism (SNP) association studies and clinical data, suggesting that risk SNPs impact on hippocampal structure and function. We hypothesized that altered expression of DISC1 and/or its molecular partners (nuclear distribution element like [NUDEL], fasciculation and elongation protein zeta-i [FEZ1], and lissencephaly 1 [LIS1]) may underlie its pathogenic role in schizophrenia and explain its genetic association. We examined the expression of DISC1 and its binding partners in the hippocampus and dorsolateral prefrontal cortex of postmortem human brains of schizophrenic patients and controls. We found no difference in the expression of DISC1 mRNA in schizophrenia, and no association with previously identified risk SNPs. However, the expression of NUDEL, FEZ1, and LIS1 was significantly reduced in tissue from schizophrenic subjects, and the expression of each showed association with high-risk DISC1 polymorphisms. These data suggest involvement of genetically linked abnormalities in the DISC1 molecular pathway in the pathophysiology of schizophrenia. PMID- 17117618 TI - [Introduction and calibration of the Chinese sun hazemeter network]. AB - Much of the current uncertainty in the quantitative assessment of the climate and environment change is due to our lack of knowledge of the aerosol, for which the large-scale sun hazemeter ground-based network directly provides basic data. The data also can revise the results of the satellite remote sensing. Depending on the stations of CERN, the standard network was first built in China. The accurate and reliable hazemeter (LEDs) was uniformly used in the network. The hazemeters were calibrated by Langley plot method and transfer calibration and were uniformly checked. The AODs, which were synchronously retrieved from hazemeters (RSD < 3%) and CEMIL (RSD < 5%), were coherent and comparable. The stability and reliability of the hazemeters and the network was approved. PMID- 17117619 TI - [AOD and angstrom parameters of aerosols observed by the Chinese sun hazemeter network from August to December 2004]. AB - Atmospheric aerosol optical depth (AOD(lamda=500 nm)), Angstrom turbidity coefficient (beta) and Angstrom wavelength exponent (alpha) are obtained using the CERN sun hazemeter network from Aug to Dec, 2004. The results are as follows: At the Tibetan Plateau, Haibei and Lhasa, the mean of AOD is 0.09, 0.12; the mean of beta is 0.05, 0.13; the mean of a is 1.09, 0.06, respectively. At the Northeast of China, Hailun and Sanjiang, the mean of AOD is 0.14, 0.15; the mean of beta is 0.04, 0.06; the mean of a is 2.32, 1.58, respectively. At the desert region of North China, e.g., Fukang, Shapotou and Eerduosi, the range of averaged AOD is from 0.17 to 0.32; the range of averaged beta is from 0.09 to 0.19; the range of averaged a is from 0.68.to 1.28. At the forest areas, e.g. Changbai Mountain, Beijing forest and Xishuangbanna, the range of averaged AOD is from 0.19 to 0.42; the range of averaged beta is from 0.12 to 0.19; the range of averaged a is from 1.11 to 1.25. At agriculture areas, e.g. Shenyang, Fengqiu, Taoyuan and Yanting, the range of averaged AOD is from 0.34 to 0.68; the range of averaged beta is from 0.18 to 0.38; the range of averaged a is from 0.97 to 1.39. At the littoral areas and the lake of East China, e.g. Jiaozhou Bay, Shanghai City and Tai Lake, the range of averaged AOD is from 0.49 to 0.68; the range of averaged beta is from 0.21 to 0.29; the range of averaged a is from 1.24 to 1.37. At the inland cities, Beijing City and Lanzhou City, the mean of AOD is 0.47, 0.81; the mean of beta is 0.20, 0.45; the mean of a is 1.66, 0.89, respectively. The variations of aerosol properties at nineteen stations are explained in the paper. PMID- 17117620 TI - [Study on mechanism of enhanced UV-B radiation influencing on N2O emission from soil-winter wheat system]. AB - To study the influencing mechanisms of enhanced UV-B radiation on the emission of N20 from soil-wheat system, outdoor pot experiments with simulating 20% supplemental level of UV-B were conducted. Results indicate that the enhanced UV B had no significant impact on the emission of N20 from soil-wheat system in turning- green stage, but declined the N2O flux and the rate of the system's respiration in elongation stage. The impact mechanisms of enhanced UV-B radiation on the N2O flux were to directly change the nitrogen metabolism process of wheat plant, such as significantly increasing soluble protein, total nitrogen and total phosphorus in wheat leaves. But the effects of UV-B radiation on soil N2O emission may be indirect, namely, UV-B treatment by working on wheat plant significantly increased the soil available nitrogen, soil microbial biomass C and N, and also changed the ratio of soil microbial C: N(from 5.0 to 6.8) in winter wheat rhizosphere. PMID- 17117621 TI - [Effect of SO2 on the catalytic performance of CoH-ZSM-5 for selective catalytic reduction of NO by CH4]. AB - Selective catalytic reduction of NO by methane over CoH-ZSM-5 in the presence of SO2 was studied. SO2-TPSR and NO+ (O2)-TPD techniques were applied to quantify the active sites and probe the poison effect. At 773 K, addition of 78 x 10(-6) SQ2, the NO to N2 conversion ratio decreases from 72% and reaches stable level 58%. Increasing the reaction temperature to 823 K, the NO to N2 conversion ratio increases to 62%. Co-existence of 2.55% H2O and 78 x 10(-6) SO2 at 773 K, the NO to N2 conversion ratio decreases further to 51% , but almost no effect is observed at 873 K. Three SO2 desorption centers at around 690 K, 810 K and 910 K are formed over the SO2-TPSR spectrum of the poisoned catalyst. Even at 970 K, the species contained sulfur is not desorbed completely. The amount of adsorbed NO and active intermediate--NO, species decreases over the poisoned catalyst, indicating that partial active sites are covered by the compound contained sulfur. As a result, the NO conversion ratio decreases. On the other hand, the presence of SO2 inhibits the conversion of CH4 and the compounds contained sulfur is desorbed further at higher temperature, which causes the temperature where the optimum NO conversion shifts to 823 K. PMID- 17117622 TI - [Influence of mineral matter on sulfur conversion in coal during combustion]. AB - Three species micro-pulverized coals(Hegang, Tiefa, Zhungeer coal) were studied, the mineral matters (MgO, CaO, Al2O3 and Fe3O4) were respectively added to the coals. The combustion of samples were studied to investigate the effect of mineral matter on transformation of sulfur during combustion by the combined of DTG and GC-MS, the flowmeter 50 mL/min, heating rate 20 degrees C/ min, oxygen volume percentage 20% . The SO2 release curve of primitive micro-pulverized coal appear three peaks during the combustion, but the demineralized sample appear two peaks. The species of coal has effect on temperature of the maximum release rate of SOz, the release rate of SO2 of Hegang coal is even in three temperature ranges, Tiefa coal appear maximum value about 500 degrees C and Zhungeer coal about 200 degrees C which probably due to the different amount of all kinds of sulfur in primitive coal sample. The mineral matter (MgO, CaO, Al2O3 and Fe3O4) have sulfur retention and catalyzing effect on SO2 the combustion of coal. The amount and species of mineral matter and species of coal determine the sulfur retention effect. PMID- 17117623 TI - [Short-term monitoring of methane emission regulation from a municipal solid waste landfill]. AB - To understand the regulation and influencing factors of methane emission from landfills in China, rates of methane emission from Hangzhou Tianziling municipal solid waste landfill were measured by static chamber technique. The emission rates ranged from 3.67 to 36.65 mmol x (m2 x h)(-1) on average of the two tested points (No. 1 and No. 2) during 24-hour cycle. The max ratio of emission rates between the two tested points was 625. The difference with the rates of the two tested points could be explained by the depth of the cover soils. Rate of methane emission from each point correlated with atmospheric pressure and water content of landfill cover soils, and the result from stepwise regression was r2 = 0.89 for point No. 1, r2 = 0.76 for point No. 2. Rates of methane emission varied from -0.23 to 0.32 mmol (m2 x h)(-1) on average in 40-day test (points with same depth of cover soils). The low correlation (r2 = 0.15) with atmospheric pressure showed that atmospheric pressure was not the decisive factor. For some time, emission rates correlated with water content or temperature of the landfill cover soils significantly, but not obvious (r2 < 0.4) during the whole monitoring time. When atmospheric pressure was included, the results of stepwise regression were improved significantly, which showed that methane emission rates were influenced by different factors, but the effect of each factor was not the same at a different time. PMID- 17117624 TI - [Distribution of polybrominated diphenyl ethers in aquatic species from the Pearl River Estuary]. AB - Polybrominated diphenyl ethers (PBDEs) were detected in biota samples collected from the Pearl River Estuary. The concentrations of sigma10 PBDEs (BDE28, 47, 66, 100, 99, 85, 154, 153, 138, 183) in fish species (including Platycephalus indicus, Pseudosiaena crocea , Pampus argenteus, Cynoglossus robustus, and Harpodon nehereus), shrimps (including Metapenaeus ensis and Metapenaeus affinis) and Squilla oratoria were from 37.8 ng x g(-1) to 407.1 ng x g(-1) (normalized to lipid), from 49.0 ng x g(-1) to 239.1 ng x g(-1) and from 142 ng x g(-1) to 444.5 ng x g(-1), respectively. BDE47 was the dominant congener in all biota samples, with a contribution to sigma10 PBDEs ranging from 53.7% to 66.9%. Differences in concentrations of PBDEs and PBDE patterns in different species may be related to the different feeding habit, living environment and uptaking or elimination rate. High ratios of concentrations of PBDEs in liver /muscle in fish species indicated that PBDEs tended to accumulate more in liver than in muscle. PMID- 17117625 TI - [Pollution of polycyclic aromatic hydrocarbons (PAHS) in middle and lower reaches of the Yellow River]. AB - Samples of water, suspended particulate matter (SPM) and sediment in the Yellow River was analyzed. The total concentration of PAHs in the main river varies from 179 ng/L to 369 ng/L (sigma 15PAHs) in water, from 54 microg/kg to 155 microg/kg (sigma 13PAHs) in SPM dry weight, and from 31 microg/kg to 133 microg/kg (sigma 13PAHs) dry weight in sediment. The levels of PAHs in water of tributaries are higher than those in the corresponding sites in the main river, and concentrations of benzo(a)pyrene in most of the stations sampled are above drinking water standard. In SPM, PAHs of 3 to 6 benzene rings are mainly correlated to the content of total organic carbon (TOC) in different sites of the main river, while only 4 to 6 rings PAHs in sediment of main river are correlated to TOC for the influence of Mengzhou Canal, whose concentration of 3-ring PAHs are quite high. The distribution of PAHs in all media sampled in the main river indicates that PAHs are mainly transported from water into SPM between Mengzhou and Jiaogong Bridge, while in the reach between Jiaogong bridge and Huayuankou a great reduce of PAHs in SPM is mainly due to the diluting effect in the interchanging process between sediment and SPM. Source analysis also reveals that PAHs are mainly originating from coal burning, although in some tributaries PAH inputs could come from combustion of petroleum. PMID- 17117626 TI - [Profiles of sediment porewater nutrient concentrations on tidal flat of turbidity maximum zone in Changjiang (Yangtze River) estuary]. AB - Profiles of nutrients (i.e., NH4+, NO2-, NO3-, PO4(3-), and SiO3(2-)) in porewater of sediment were investigated monthly in the intertidal flat of Chongming Dongtan in Changjiang (Yangtze River) estuary during March 2005 to February 2006. The results indicate that concentrations of NH4+ and SiO3(2-) in porewater are always in the range of 200 micromol/L to 500 micromol/L and show different patterns of distribution profile among high, middle, and low marshes. Compared to high and middle marsh, low marsh, without vegetation coverage and of coarser grain size, generally contains higher NH4+ and lower SiO3(21) concentrations. SiO3(2-) concentrations are predominantly determined by seasonal temperature change, but variations of NH4+ are more complicated and prone to exhibit lower values in spring and summer when the vegetation above show relatively high growth rate. Concentrations of NO2- + NO3- and PO4(3-) in porewater are commonly much lower than NH4+ and SiO3(2-) by 2 to 3 orders of magnitude, however, due to factors such as bioturbation, the concentrations in top sediment may be higher than 10 micromol/L. The relatively high NO3- concentration in top sediment,together with the rapid transfer from oxygenic to anoxic environment, make it possible that significant denitrification happens, which may draw its remarkable influence on the biogeochemical cycles of nutrient elements in the land-sea interaction area in Changjiang (Yangtze River) estuary. PMID- 17117627 TI - [Characterization of urban surface runoff in two urban catchments in Macau]. AB - Characteristics of pollutants from urban surface runoff were investigated. Two catchments with a seperated system in Macau were selected for sampling on rainfall events during the period of August to November, 2005. Water quality parameters such as pH, turbidity, TSS, COD, TOC, TN, TP, Zn, Pb, and Cu were analyzed. The results show that the commercial-residential urban catchment shows high level of COD, TN, TP, and the park urban catchment has high TN and TP concentration. From the pollutograph analysis, the peak of TSS, TN, TP concentration appears in the first and the third sample respectively in ELH and YLF catchments, and then the pollutants concentration tend to decrease. Regression analysis between TSS and TN & TP in two urban catchments resulted in a high value (R2 > 0.95) of the coefficient of determination R2 indicating a close relationship between soil losses and nitrogen & phosphorus discharged from surface runoff. The profile of TSS and COD discharged from surface runoff relates greatly to the surface flow change, whereas the surface flow change has little influence on the profile of TN and TP. The heavy metals such as Zn, Pb and Cu fluctuate with the continuous input of vehicles during rainfall events. Pollutants such as TSS, TN, COD discharged from surface runoff depend greatly on the dry periods and storms intensity in such two urban catchments. PMID- 17117628 TI - [Sustainable operation of subsurface constructed wetland treating polluted river water]. AB - Water quality, flux and temperature of polluted river in northern China fluctuate a lot in a year around, which is disadvantageous for sustainable operation of subsurface constructed wetland treating polluted river water. A pilot system of subsurface constructed wetland treating polluted river water was operated for one year to study the sustainable operation of the process. The results show that ammonia removal performance fluctuated a lot according to different seasons. At the hydraulic loading of 15 cm/d, ammonia nitrogen removal rates were above 70% in summer, however, below 30% in winter when water temperature was below 15 degrees C. Changes of water temperature had little influence on COD removal performance. In the wet season of summer, the pilot wetland system could bear a short-period flood hydraulic loading, and ammonia nitrogen and COD removal rates were 52% and 36% respectively at the hydraulic loading of 100 cm/d. The activity of soil dehydrogenase had a positive correlation with seasonal changes of temperature and pollution removal performance. PMID- 17117629 TI - [Research on dissolved nitrogen and phosphorus loading model based on GIS]. AB - In order to research the origin of nonpoint source pollution dissolved nitrogen (DN) and dissolved phosphorus (DP), some artificial rainfall experiments was made at Guanting Reservoir. The experimental data show that the correlation between transporting rate of DN (DP) and the runoff is very good, with the average correlation coefficient 0.9978 and 0.9889 for DN and DP respectively. Therefore, a new load model of DN(DP) is put forward. While applying it, we studied the spatial distribution of DN (DP) pollution load according to the digital elevation model (DEM) and the information obtained from land use map and soil map. Its results show that DN mainly originated from irrigable land first, mound and hill second. PMID- 17117630 TI - [Evaluating eutrophic state of Taihu Lake by in situ hyperspectra]. AB - In situ experiments including water quality analyzing and spectra measuring were conducted on Jun., 2004 and Aug., 2004 in Taihu Lake for 21 samples. Then eutrophic state of Taihu Lake was evaluated by measured hyperspectra data, and the evaluating index was eutrophic state index (TSI). Firstly, the simulation model of water reflectance was built by analytic model using inherent optical properties (IOPs). Then Chlorophyll a content (Chl-a) was inversed by optimizing method by Matlab software; Secondly, TSI was calculated by inversed Chl-a, and the eutrophic state of Taihu Lake was evaluated by using ArcView' s interpolation function. The evaluation figures show that: there are notable differences between Jun., 2004 and Aug., 2004. The proportion of mesotropher state is as high as 61% in June, while the proportion of light eutrophic is as high as 52% in August. In totally, the most serious eutrophic state occurs in north of Taihu Lake. Eutrophlic state wears off from north to south. The lowest eutrophic region is in the east of Taihu Lake. PMID- 17117631 TI - [Nitritation-ANAMMOX process for treatment of ammonium rich wastewater]. AB - Combination of a nitritation process and anoxic ammonium oxidation (ANAMMOX) process for the treatment of ammonia rich influents is evaluated. Herein the combined process was studied with manmade synthetic wastewater. Research results show that when the hydraulic retention time (HRT) was I day, the nitritation activity was stable and the influent ammonium concentration has little effect on it. When the influent ammonium concentration is 400-600 mg/L, the effluent nitrite concentration is always between 260 mg/L and 280 mg/L, so the effluent nitrite/ammonium ratio can be adjust by control the influent ammonium concentration. The total nitrogen elimination was depended strongly on the nitrite/ammonium ratio in the inlet of the ANAMMOX reactor. When the inlet ammonium was 480 mg/L of the combined process, the nitrite/ammonium ratio of the nitritation process was about 1.2, the total nitrogen elimination was 84%. PMID- 17117632 TI - [Study on denitrification characteristics of dynamic membrane based on nitrate liquid-membrane microelectrodes]. AB - Nitrate microelectrodes and ORP microelectrodes were fabricated to study the denitrification characteristics of dynamic membrane at different COD loadings. The denitrification process was found at 0.6-1 mm depth beneath the interface of biofilm/bulk. The results of ORP microelectrode also demonstrated that the ORP value in the range of denitrification area was between 88.6 approximately -128.4 mV which was appropriate to denitrification. When the COD loading was 0.45 kg/(m3 x d), the denitrification rate (NO3- -N) was the maximum of 0.6347 x 10(-6 mol/(L x s). With the increase of COD loading, the denitrification area was increasing and two layers with different denitrification rates emerged in the dynamic membrane. The phenomenon implied the effect of organic concentration, oxygen concentration and bacterial competition on the denitrification rate. PMID- 17117633 TI - [Biological wastewater treatment and simultaneous generating electricity from organic wastewater by microbial fuel cell]. AB - An air-cathode microbial fuel cell (ACMFC) was successfully started up using anaerobic activated sludge as inoculums, generating a voltage of 0.24V after inoculations for 110 h. When using acetate and glucose as substrate, voltage of 0.38V and 0.41V (based on external resistance of 1000 omega) is obtained; meanwhile, the maximum power density reaches 146.56 mW/m2 and 192.04 mW/m2 respectively, suggesting that organic wastewater can be used to produce electricity. Removal efficiency of 99% (acetate) and 87% (glucose) is achieved simultaneously, demonstrating that ACMFC can treat organic wastewater. Electron recovery efficiency as low as 10% for both acetate and glucose is observed mainly due to aerobic respiration of microorganisms caused by diffusion of oxygen molecular from the cathode, leading to electron loss. MFCs are capable of converting chemical energy presented in organic wastewater into electricity energy with accomplishments of wastewater treatments simultaneously, which possibly captures considerable benefits in terms of environments and economics. PMID- 17117634 TI - [Effect and mechanism of degradation of nitrobenzene in aqueous solution by O3/H2O2]. AB - Nitrobenzene (NB) was selected as the model pollutant in water and the efficiency and mechanism of degradation of NB in aqueous solution by O3/H2O2 were investigated. The effects of pH, H2O2 dose and the inhibitor or accelerant of .OH on the removal rate of NB were studied. H2O2 could obviously improve the ozonation decay rate of NB when the pH value of the solution was below 7. The removal rate of NB was enhanced remarkably while H2O2 dose was increased from 1.0 m g/L. to 4.0 mg/L. However, as H2O2 dose increased from 4.0 mg/L to 20 mg/L, the removal efficiency of NB decreased. Different quantities of H2O2 were yielded in different reaction phases of single ozonation system. Both systems of single ozonation and H2O2-catalysed ozonation could not reduce TOC observably. During the NB degradation process, organonitrogen was almost completely converted to nitrate and the pH value of the solution reduced significantly. Results of LC-MS and GC-MS analysis showed that the main intermediate products were phenolic compounds and carbonyl compounds. A possible reaction pathway of the catalytic ozonation of NB was also proposed. It was found that the catalytic ozonation of NB could be divided into two steps. First, hydroxyl radical attacked phenyl ring to form phenolic compounds, then the ring was opened, forming into various aliphatic compounds or being mineralized to inorganic compounds. PMID- 17117635 TI - [Effects of potassium permanganate on natural organic matter chlorination activity]. AB - Effects of potassium permanganate (PP) oxidation on natural organic matter(NOM) chlorination activity was studied in the article. Natural organic matter was separated into humic acid (HA), fulvic acid(FA), hydrophilic acid(HPIA) and non hydrophilic acid (HPI-NA) fraction four fractions by XAD resin adsorption technique. Potassium permanganate oxidation alone increased HA and HPIA chlorination activity and decrease that of FA and HPI-NA. The chlorination activity of HA and HPIA were increased by 39.3% and 13.8% by 0.75 mg/L. potassium permanganate, but the chlorination activity of FA and HPI-NA were decreased by 33.6% and 46.9%. SUVA results showed that potassium permanganate increase FA and HPIA unsaturated band contents, and decreased that of FA and HPI-NA. Compared with coagulation, potassium permanganate combined with coagulation can decrease the trihalomethanes of HA, FA, HPIA and HPI-NA by 9.1,15.7,7.2 and 14.7 per cent compared with that of coagulation alone. PMID- 17117636 TI - [Reductive dechlorination of chlorinated hydrocarbons in water by Ag/Fe catalytic reduction system]. AB - Dechlorination of many kinds of chlorinated hydrocarbons including trichloromethane (CF), tetrachloromethane (CT), 1, 1,1-trichloroethane (1, 1, 1 TCA), 1, 1, 2, 2-tetrachloroethane (1, 1, 2, 2-TeCA), hexachloroethane (HCA), trichloroethylene (TCE), perchloroethylene (PCE) in water by Fe and Ag/Fe were investigated. Results show that the existence of Ag on Fe can enhance the dechlorination rate of chlorinated hydrocarbons. When the surface area concentration of iron is 150 m2 x L(-1), the dechlorination rate constant of CF, CT, 1, 1, 1-TCA, 1, 1, 2, 2-TeCA, HCA are 0.084 h(-1), 2.358 h(-1), 0.417 h(-1), 0.215 h(-1), 1.098 h(-1) by single Fe0, in this condition, no dechlorination of TCE and PCE during 10h, while the dechlorination rate constant of CF, CT, 1, 1, 1 TCA, 1, 1, 2, 2-TeCA, HCA, TCE, PCE are up to 1.850 h(-1), 9.504 h(-1), 1.624 h( 1), 1.778 h(-1), 2.842 h(-1), 0.463 h(-1), 1.251 h(-1) by Ag/Fe. The dechlorination rate constant of CF can increase by the factor of 20. The reaction pathways of chlorinated hydrocarbons with Fe and Ag/Fe were mainly including hydrogenolysis, reductive elimination and dehydrochlorination. The hydrogenolysis is the main reaction pathway for CF, TCE and PCE. The dechlorination pathway of 1, 1, 1-TCA are both hydrogenolysis and dehydrochlorination. Reductive beta elimination to form DCE is the dominant reaction pathway of 1, 1, 2, 2-TeCA, and HCA is via reductive beta elimination to form PCE firstly, then via hydrogenolysis to DCE. PMID- 17117637 TI - [Microwave assisted photocatalytic degradation of phenol in aqueous solution]. AB - Experimental study on microwave assisted photocatalytic degradation of phenol in aqueous solution was carried out with modified domestic microwave oven, electrodeless discharge lamps (EDLs) and TiO2 catalyst. Results showed that the removal rate of phenol in the MAPC process was 92% after 30 min reaction. Loss of total organic carbon (TOC) was 84% . The optimal conditions for the MAPC method were 10 mg/L phenol, 900 W MW output energy, 50 mL solution volume, EDLs-3, 1-4 g/l. catalyst dosage and 15 mL/min of flowing velocity in the circulating mode. The MAPC process was promising in treating phenolic waste water. PMID- 17117638 TI - [Photocatalytic degradation of formaldehyde and VOCs in air on the porous nickel mesh coated with nanometer TiO2]. AB - Three different metal ions doped TiO2 photocatalysts, which were prepared by the sol-gel method, were immobilized to porous nickel mesh by coating. The photocatalytic degradation activity of the supported photocatalyst on formaldehyde and volatile organic compounds (VOCs) was investigated. The results show that the nanometer TiO2 has an anatase structure. The photocatalytic degradation rate of formaldehyde and VOCs of 1.5% La3+ doped TiO2 coated on porous nickel mesh at 90 min are: 94% and 87%, higher than undoped TiO2: 83% and 72%, Fe3+ doped TiO2: 62% and 62%, Ag+ doped TiO2: 86% [Chinese character: see text] 81%. The orders of photocatalytic degradation rate on formaldehyde and VOCs with different content of La3+ doped TiO2 are as follows: 1.5% > 1% > 2% > undoped, 1.5% is the optimum La3+ doped content. Decreasing circular wind speed and using 254 nm or 365 nm ultraviolet wavelength will not influence the photocatalytic degradation rate of formaldehyde and VOCs. PMID- 17117639 TI - [Comparative study on accumulation of Ah-receptor agonists in contaminated soil based on EROD bioassay and chemical analysis]. AB - The potential ecological risk by wastewater or reclaimed water for irrigation is of great concerns in recent years, but little work was done on the chronic toxicities through long term accumulation of persistent organic chemicals in soil. In present work, concentration of Ah-receptor agonists in soil organic extract was measured by an ethoxyresorfin O-deethylase (EROD) bioassay, which was calibrated and expressed by the 2, 3, 7, 8-tetrachlorodibenzo- p-dioxin (2, 3, 7, 8-TCDD) toxic equivalent (TEQbio). Simultaneously, 16 PAHs in soil were analyzed and their TEQs (total as TEQ(PAHs)) were calculated according to their toxic equivalent factors (TEFs) cited from literature. By bioassay, it was found that the concentration level of Ah-receptor agonists in soil irrigated using reclaimed water could be as high as 97.4 ng/kg, which was obviously higher than that in background soil using ground water irrigation regime (56.0 ng/kg). In comparing the results from bioassay and chemical analysis, the percentage of TEQ(PAHs) in TEQbio increased from 10.3% in background soil to 78.6% in the soil irrigated by reclaimed water. Use of reclaimed water for irrigation could result in the accumulation of Ah-receptor agonists in soil,and a major part of them in this case could be attributed to the accumulation of 16 priority PAHs in soils. PMID- 17117640 TI - [Evaluation of estrogenic activity for nonylphenol using quantitative real-time RT-PCR method]. AB - It has been demonstrated that nonylphenol (NP) exerts estrogenic activity. Quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) was used to study the VTG-I , VTG-II , CHG-H and CHG-L genes expression in the liver of juvenile medaka exposed to NP at 1, 10, 50, 100 microg/I. for 60 days. The results show that the VTG-I , VTG-II, CHG-H and CHG-L genes expression in the liver of juvenile medaka are induced even at 1 microg/L, significantly. It should be noted that the lowest-observed-effect concentration (LOECs) based on the hepatic vitellogenin (VTG) induction is about 1 microg/L, suggesting that quantitative real-time RT-PCR can detect the estrogenic activity of NP at relatively low concentration, and there is a potential application in evaluating the estrogenic activity of NP in aquatic environment. PMID- 17117641 TI - [Flow injection biosensor based on the immobilized AChE]. AB - A biosensor based on the flow injection system was constructed with the immobilized AChE from Scomberomorus niphonius (Curier) as identification element and a pH electrode as transducer. When phosphate buffer was used as carrier liquid, a good reproducibility (RSD = 1.427% , n=10) of the biosensor response was obtained after the substrate was injected repetitively. After an incubation time of 20 min, the calibration graph to methyl-parathion is linear (r = 0.9986) when its concentration ranges from 4.29 x 10(-10) mol x L(-1) to 4.29 x 10(-8) mol x L(-1) , and the detection limit is 1.3 x 10(-10) mol x L(-1). However, the sensitivity of this biosensor to methyl-parathion when using clean seawater as carrier liquid is not as good as that using phosphate buffer. But after preoxidation of methyl-parathion with NaClO as oxidant, the detection limit of the biosensor to methyl-parathion in seawater can be improve to 2.16 x 10(-7) mol x L(-1). PMID- 17117642 TI - [Degradation pathway of 17alpha-ethynylestradiol by Sphingobacterium sp. JCR5]. AB - A bacterial strain, JCR5, which degrades 17alpha-ethynylestradiol (EE2), was isolated from activated sludge of wastewater treatment plant treating wastewater from pharmacy factory mainly producing contraceptive medicine in Beijing, China. Based on its morphology, physiology biochemical properties and 16S rDNA sequence analysis, this strain was identified as Sphingobacteriumn sp. JCR5. Strain JCR5 can grow on EE2 as sole carbon and energy source. The degradation process for EE2 with initial concentration of 30 mg x L(-1) was studied, and the result indicated that the degradation rate of EE2 within 10 days was 87%. Experiments of different substrates showed that strain JCR5 can grow on many substrates other than EE2 such as several steroidal estrogens (estrone, 17beta-estradiol, estriol and mestranol), the intermediates in contraceptive medicine processing and some aromatic compounds. Mass spectrum analysis demonstrated that EE2 was oxygenized to estrone (E1) firstly, and 2-hydroxy-2,4-dienevaleric acid and 2-hydroxy-2,4 diene-1, 6-dioic acid were the main catabolic intermediates during EE2 degradation. The former adopted a pathway that was analogical to the pathway of the previously reported testosterone-degrading Comamnonas testosteroni TA441, and the latter was a metabolite with a different cleavage position of 3-hydroxy-4,5 9, 10-disecoestrane-1(10), 2-diene-5,9, 17- trione-4-oic acid from the former. PMID- 17117643 TI - [Biodegradation of benzene series compounds under nitrate reducing conditions]. AB - A series of batch experiments were performed using mixed bacterial culture to investigate the biodegradability of benzene, toluene, ethylbenzene and the xylene isomers (BTEX) under denitrifying conditions. The results show that BTEX can be degraded effectively by the mixed bacterial culture. The biodegradation of BTEX could be modeled by the substrate inhibition Monod equation. When the concentrations of BTEX were less than 50 mg x L(-1), the anaerobic degradation rates decreased with toluene > ethylbenzene > m-xylene > o-xylene > p-xylene > benzene. The ratio between the amount of nitrate consumed and the amount of the benzene, toluene, ethylbenzene, o-xylene, m-xylene, p-xylene degraded was 9.47, 9.26, 11.14, 12.46, 13.36 and 13.02, respectively. The electron acceptor needed in the initial anaerobic degradation of BTEX was nitrate, and then both nitrate and nitrite could be served as the electron acceptors for the further degradation of the intermediates of BTEX. PMID- 17117644 TI - [Relationship between several biologic indices and degradation efficiency of bioaugmentation system]. AB - Because of the existence of some unstable factors, the effluent water from a bioaugmentation refinery wastewater treatment system usually exceeds the provided standard. To solve this problem, the relationship between the bacteria number, dehydrogenase, catechol 1,2-dioxygenase(C12O), catechol 2,3-dioxygenase(C23O) and the degradation efficiency of the bioaugmentation and control system is investigated and discussed. Using ERIC-PCR technology, we studied the changes of the microbial populations before and after treatment. The results show addition of microorganisms agent can increase the bacteria number, enzyme efficiency and the efficiency of the reactor. There exists a positive relationship between the bacteria number, C23O and the degradation efficiency in the bioaugmentation reactor. The microbial populations in both the reactors are stable before and after run. C12O is a kind of induction enzyme and its efficiency declines while the degradation efficiency increases. PMID- 17117645 TI - [Community structure of nitrification bacteria in aerobic short-cut nitrification granule]. AB - By using a lab-scale aerated upflow sludge bed reactor, the inoculated anaerobic granule was cultivated to aerobic nitrification granule, and then converted to short-cut nitrification granule with the short-cut nitrification efficiencies above 90%. Appling real-time quantity PCR, and florescent in situ hybridization techniques, the ecological community structure of nitrification bacteria in aerobic granules were studied. The results show that there existed a layered structure in the aerobic granule, the ammonia oxidizing bacteria (AOB) was mainly located in the surface area of the granule, and the nitrite oxidizing bacteria (NOB) was mainly located in the inner area of the granule, was just adjacent to the AOB. There was no active bacteria in the inner core area. The amount of AOB in the granules increased, as the ammonia loading rate of the reactor was increased gradually. The percentage of AOB in the total amount of Eubacteria in the granule was 0.45%, 5.20%, 15.37%, 48.55% respectively, as the ammonia loading rate of the reactor were 0, 0.4, 1.0 and 2.2 kg/(m3 x d) respectively, and the nitrosofication efficiency were 0%, 35%, 50%, 99% relatively. PMID- 17117646 TI - [Effects of hydraulic retention time (HRT) on nitrification performance and microbial community of conventional activated sludge (CAS)]. AB - A conventional activated sludge system was used to treat the ammonium-bearing inorganic wastewater for 260 days under decreased HRTs. When treating 500 mg x L( 1) NH4+ -N wastewater, over 98% ammonia removal could be obtained at HRT > or =20 h. Sharp washout of biomass occurred when HRT was further decreased, resulting in the accumulation of ammonium and nitrite. Changes in the community structure were monitored by microbial quinone and conventional analytical methods. Quinone profiles indicated that ammonia-oxidizing bacteria (AOB) in beta-Protevbacteria and Nitrobacteria in alpha-Proteobacteria were the predominant species to oxidize ammonia and nitrite, respectively. The number of AOB decreased with the reduction of biomass in bioreactor, while that of nitrite-oxidizing bacteria (NOB) changed little and fluctuated at 10(5) CFU x L(-1) PMID- 17117647 TI - [Soil-based eco-toxicity of petroleum to terrestrial higher plant after phytoremediation]. AB - Petroleum-contaminated soil after five-year phytoremediation was taken as tested soil initially spiked with a serial diesel concentration of 5 000, 15 000 and 30 000 mg/kg (dry weight). Residual concentrations of mineral oil by chemical analysis of gravimetry, as well as the soil-based eco-toxicity to wheat (Triticum aestivum L.), the terrestrial higher plant by several ecotoxicological bioassays including seed germination and root elongation test, early seedling growth test, contents of cytochrome P450 monooxygenases, activities of antioxidant enzymes (superoxide dismutase, SOD, and peroxidase, POD), and lipid peroxide (malondialdehyde, MDA content in wheat seedling leaves, etc. were evaluated. Results showed that mineral oil was well removed in each treatment by chemical analysis, with residual concentrations ranging from 199 to 877 mg/kg (dry weight) and with total removal rates between 90.1% and 97.2%. The evaluating results by eco-toxicological assays differed to some extent from those by chemical analysis, meanwhile, eco-toxicity of each treatment differed depending on endpoints by different bioassays. Among the ecotoxicological indexes, root length (48 h), root fresh weight (7 d), contents of P450, activities of SOD, and contents of MDA, etc. exhibited better indication to the soil toxicity. The general evaluation by combining the two analyses chemical and eco-toxicological indicated that the ecological risk was higher in most intermediately- and heavily-contaminated treatments (initially spiked with diesel concentration of 15,000 mg/kg and 30,000 mg/kg). PMID- 17117648 TI - [Effect of elevated atmospheric O3 on arbuscular mycorrhizaL (AM) and its function]. AB - In order to understand the possibility of AM in protecting crops from O3 pollution, simulated experiment was carried out to investigate effect of increased atmospheric O3 (0.02 microL x L(-1), 0.1 microL x L(-1) and 0.2 microL x L(-1)) on AM and its function. The results indicate that mycorrhizal colonization was slightly affected by elevated atmospheric O3, but spore and hypahe growth were significantly impacted. Spore number at high O3 level increased I times than that at ambient O3; hypahe length of AM at low and high O3 level decreased 48.7% and 85.6% than that at ambient O3, respectively. Root biomass, nodule number, horsebean pod and grain weight (excluding the treatment-- S + M) significantly decreased with elevated atmospheric O3, but they markedly increased after inoculation with AM fungi. Compared with the uninoculated treatment, nodule number increased by 2-5 times and grain weight of horsebean increased by 3-6 times after inoculation with AM fungi. Relative function of AM was slightly impacted by elevated atmospheric O3. Although indigenous microbes competed with inoculated AM fungi, inoculation with AM fungi could decrease the stress of O3 on plant, which was positive significant to alleviate crop loss resulted from O3 pollution. PMID- 17117649 TI - [Effect of limestone and magnesite application on remediation of acidified forest soil in Chongqing, China]. AB - Effect of limestone and magnesite application on remediation of a typical acidified soil under a masson pine (Pinus massoniana) forest at Tieshanping, Chongqing in southwest China was studied through field experiments. The changes of soil water chemistry in different layers within one year after application of limestone or magnesite indicated that the remediation agents leaded to the recovery of acidified soil by significant increase of pH value and concentration of relative cation, i.e., Ca2+ or Mg2+, and notable decrease of inorganic monomeric aluminum (Ali). However, the accelerated leaching of NO3- and SO4(2-) might somewhat counteract the positive effects. Since the limestone powder applied was much finer and thus more soluble than the magnesite powder, it seemed that the addition of limestone was more effective than that of magnesite. However, the application of magnesite could probably improve the nutrient uptake and growth of plant, and thus limestone and magnesite should be used together. The change of soil water chemistry was much more notable in upper layer of soil than lower, which means that it will take long time to achieve the whole profile soil remediation. PMID- 17117650 TI - [Impact of dissolved organic matter on plant uptake of phenanthrene and its mechanisms]. AB - Hydroponic assays were conducted to investigate the influence of dissolved organic matter on uptake of phenanthrene by wheat as well as its mechanisms. The results showed that, under hydroponic condition, phenanthrene impairment of plant growth occurred with wheat growth inhibited rate of 18.01%. The impairment would be greatly enhanced in the presence of dissolved organic matter (DOM) derived from pig manure, and the inhibited rate increased to 24.38%. Wheat could uptake and accumulate phenanthrene in the nutrient solution, which could be escalated by DOM, as indicated by wheat root bioconcentration factor being increased to 37.63 L x kg(-1) in the presence of DOM from 2.84 L x kg(-1) in the absence of DOM. At the same time, DOM could facilitate phenanthrene translocation from plant roots to the upper. As a result, the pH value of nutrient solution could increase by more than 1 unit when the co-existence of DOM and phenanthrene occurred in solution, suggesting that H+ -phenanthrene cotransport system is involved in the uptake of phenanthrene by plants. A synergism was also found between wheat uptakes of phenanthrene and inorganic nutrients, Moreover, DOM accelerated markedly the synergism. It is concluded that DOM affects the uptake of phenanthrene by plants and the environmental behaviors of phenanthrene. PMID- 17117651 TI - [Adsorption of toluene and naphthalene on Beijing soils and its influence factor]. AB - Static adsorption experiments were carried out to investigate the adsorption of toluene and naphthalene on Beijing soils, and to determine the influence of temperature and surfactants on adsorption. Although the affinity of petroleum hydrocarbons for seven soils is different, the adsorption isotherms of toluene and naphthalene on seven soils are well described by the Freundlich type. The adsorbed amount of naphthalene is higher than that of toluene on the same soil. High temperature doesn't benefit adsorption. Cetylrimethyl ammonium bromide (CTAB) and sodium dodecyl benzene sulfonate (SDBS) both have a positive effect on desorption of toluene and naphtalaene from soil. The desorption rate of toluene and naphthalene in the presence of CTAB and SDBS is 27.5%, 12.1% and 64.3%, 48.8%, respectively, which show that SDBS plays a much more important role in desorption than CTAB. The adsorbed quantities of toluene and naphthalene decrease with increasing SDBS concentration. When CTAB concentration is lower than the critical micelle concentration (CMC), adsorption increases with increasing CTAB concentration; When CTAB concentration is equal to or higher than CMC, adsorption decreases with increasing CTAB concentration. PMID- 17117652 TI - [Study on recovery of copper from waste printed wiring boards by electrostatic separation]. AB - Recovery of Cu from waste printed wiring boards (PWBs) was studied by crushers and electrostatic separator. The results of pulverizing experiment show that liberation degree of metal is higher in grain size -0.9 + 0.074 mm, the product of this grain size can be used as raw material in electrostatic separation. The main factors that influence electrostatic separation are found by dynamic analysis of particles in course of electrostatic separation, it conclude electrode voltage, roller rotate speed, electric coronary polar distance and granularity of particles. The -0.9 + 0.074 mm pulverizing product is separated through single stage electrostatic separation after optimizing influence factors. Result shows that the enrichment condition of Cu in fine material is good, Cu could be concentrated from 32.0% to 63.6%, and recovery ratio is 78.7%. PMID- 17117653 TI - [Study on redox zones of landfill leachate plume in subsurface environment]. AB - A soil column filled with sandy soil was constructed to investigate biogeochemical process of leachate pollutants degradation and redox zones in subsurface environment, and variation of Fe3+, Fe2+, oxidation capacity (OXC) and reduction capacity (RDC) of fresh and contaminated soil were analyzed. Experimental results indicated that four sequent redox zones are appear in pollution plume, and different terminal electron acceptors (they are CO2, Fe3+, NO3- and O2) were used by microbes in different zones; Zones can be named methanogenic zone, iron reduction zone, nitrate reduction zone and oxygen reduction zone correspondingly; the indicator of the different zone is HCO3-, Fe2+, NO2- and DO, and its highest concentration is 10,353 mg/L, 13.61 mg/L, 0.097 mg/L and 5.8 mg/L respectively. OXC of the soil is increased sequentially, its lowest value is 14.18 micromol/g in methanogenic zone and highest value is 24.45 micromol/g in oxygen reduction zone; RDC of the soil is reduced sequentially, its highest value is 14.18 micromol/g in methanogenic zone and lowest value is 24.45 micromol/g in oxygen reduction zone. With the more serious contamination, Fe3+ minerals of the soil are reduced by biogeochemical process, and dissolved Fe2+ is produced. Some of the Fe2+ is released to water and the other is precipitated as sediments. Therefore, it is significant for the in situ remediation and risk assessment of the landfill contaminated site to investigate of the redox zones. PMID- 17117654 TI - [Impact of hydrothermal process on the dewaterability and degrease performance of restaurant garbage]. AB - In order to ameliorate the dewaterability and degrease performance of restaurant garbage, and to improve the treatment effect, a complete trail series with 2 factors on 5 levels was implemented. The 2 factors were temperature and retention time respectively as the main influencing factors of hydrothermal process. By means of analyzing the variation of the resistance, dewatering rate and floatable oil content of the treated restaurant garbage, and constructing the solid grease extracting kinetics, the mechanism of impact of hydrothermal process on the dewaterability and degrease performance of restaurant garbage was studied. It showed that the dewaterability of the product decreases at the beginning, after heating for 40min, it begins to increase. Moreover, it increases more quickly as temperature increases. The optimal dewaterability of the treated garbage appears at 180 degrees C and heating for 100 minutes. As temperature rises and heating time increases, the degrease performance is improved. Furthermore, this trend becomes more remarkably as temperature increases. When the temperature and retention time reach 160 degrees C and 80 minutes respectively, most of the solid grease in the garbage is extracted out to become floatable oil which can be separated and recovered readily. Subsequently, the amount of floatable oil begins to decrease since chemical reactions such as the partial hydrolysis of the oil take place. Additionally, the extraction of solid grease from interior accords with first-order reaction dynamic model. PMID- 17117655 TI - [Exposure assessment of various reclaimed water uses]. AB - The exposure assessment method and model of various reclaimed water uses are built combining with Beijing reclaimed water project. Firstly the daily ingesting dose and lifetime average daily dose (LADD) of exposure people are provided via field work and monitoring analysis, which could be used in health risk assessment as quantitative reference. Take park irrigation as a example, for occupational workers, the total daily ingesting dose is 0.07 L/d, LADD of disinfection by products(DBPs) via the respiratory route is 2.8 x 10(-7) - 1.2 x 10(-5) mg x (kg x d)(-1), LADD of DBPs via the dermal route is 5.8 x 10(-8) - 2.4 x 10(-6) mg x (kg x d)(-1). For common people, the total daily ingesting dose is 0.04-0.05 L/d, LADD of DBPs via the respiratory route is 1.1 x 10(-7) - 6.8 x 10(-6) mg x (kg x d)(-1). PMID- 17117656 TI - [China's environmental load and environmental depressurization analysis]. AB - Based on material flow accounting and related indicators, an indicator, domestic environmental load, is formulated to measure the aggregate environmental pressure of a nation. Combining this indicator with the gross national product, an indicator for environmental efficiency is derived. The domestic environmental load is then decomposed into the rebound effect caused by economic growth and the depressurization effect induced by the efficiency increase. A case study was carried out for the Chinese economy to investigate its domestic environmental load, environmental efficiency and the rebound and depressurization effects. Results show that the environmental efficiency of the Chinese economy increased during the study period with an annual rate of 5.6%, indicating that a certain degree of depressurization was achieved. However, the rebound effect caused by economic growth was much greater than the depressurization effect induced by the efficiency increase,resulting in a considerable increase in the domestic environmental load (the annual growth rate was 3.8%). The Chinese economy is characterized by high environmental pressure and it is hard to achieve absolute depressurization. PMID- 17117657 TI - [New agents against atherosclerosis tested. Alarming findings, ACAT inhibitors seem to have proatherogenic effects]. PMID- 17117658 TI - [LDL lowering--level and time. Maybe two equally important parameters for protection against arteriosclerosis]. PMID- 17117659 TI - [Lowering of LDL cholesterol prevents cardiovascular diseases. "Normal values" are too high--treatment time is a crucial factor]. PMID- 17117660 TI - [Can higher level of HDL cholesterol augment cardiovascular disease prevention? The combination statins-nicotinic acid increases the level of HDL and reduces the level of LDL cholesterol]. PMID- 17117661 TI - [Investigation of scientific fraud. Statements from the Swedish Research Council not sufficiently normative]. PMID- 17117662 TI - [The county council and the municipalities call to rise against the European Union directive!]. PMID- 17117663 TI - [Make a distinction between management and financing!]. PMID- 17117664 TI - [Patients' legal rights are subordinate to the resources and aims of psychiatric services]. PMID- 17117665 TI - [The significance of life in mental disease]. PMID- 17117666 TI - [That everything gets better just because it is centralized is not an axiom]. PMID- 17117667 TI - [A tsunami case report--additional comment]. PMID- 17117668 TI - [Head of psychiatric service--a position or a vocation?]. AB - An attempt to create a social and psychological image of contemporary psychiatric department leader has been made. Professional and social activity of a modern head physician, his personal, business and other qualities are described. Taking into account these characteristics, the author suggests a system of competitive selection for this position and a broad discussion of the problem in psychiatric society. PMID- 17117669 TI - [The diagnosis of cerebral meningiomas at the stage of early clinical manifestation]. AB - Presented are the results of the analysis on efficacy of cerebral meningiomas diagnosis at the stage of early clinical manifestation. The case-history data of 277 patients (95.5%) examined at the early stage of clinical manifestation have been analyzed. Type and frequency of referrals to the doctor at different periods of the disease and their dynamics, the set of curative and diagnostic procedures used for patients' examination during the referrals and timeliness and degree of diagnosing intracranial meningiomas at different stages of examination have been established. Factors responsible for the clinical manifestation of the existing tumor have been determined. Diagnosis of cerebral meningiomas at the stage of early clinical manifestation remains unsatisfactory: 24.6% of patients (68) did not seek medical aid despite the presence of clinical symptoms of the tumor and 40.9% attended the doctor repeatedly. In primary examination, cerebral meningioma was diagnosed in 30.7% of outpatients and in 47.3% of in-patients. PMID- 17117670 TI - [Psychotic forms of atypical autism in children]. AB - The aim of the study was to determine clinical borders of psychotic forms of atypical autism in children, its psychopathological and age-specific manifestations as well as nosological peculiarities and to specify its pathogenetic features. Eighty patients with childhood endogenous autism, Rett syndrome, fragile X syndrome, Down syndrome have been studied during 14 years. The study showed that psychoses similar by symptoms and course, which are characterized by attacks and regressive-catatonic disorders, may develop in the course of atypical autism. These psychoses develop on the background of dysontogenesis with consequent replacement of the following stages: autistic, regressive, catatonic, with returning to the autistic stage between attacks. Psychopathological similarity of these psychoses in different disorders correlated with EEG changes of the same type (appearance of the marked I-rhythm at the regressive stage of psychosis). PMID- 17117671 TI - [Micropolarization of the brain: a noninvasive method for correction of morphological and functional disturbances in acute focal brain lesions and their consequences]. AB - The paper is devoted to the use of small direct current in correction of morphological and functional disturbances of the human brain. Two hundred and one patients aged from 7 to 82 years have been studied. In patients with focal brain damages at the acute stage (1-2 days after stroke), the anode and cathode were placed in the projection of a damaged center. In patients in "autonomic status" condition, the anode was placed both in frontal and parietal projection of the right hemisphere cortex and the cathode--on a mastoid of the right hemisphere. Strength of the current used was 300-500 mcA, time of one procedure--30-40 min. The whole treatment course involved no more than 15 procedures. Before the treatment, after 3-5 procedures of micropolarization and at the end of the treatment course, patients underwent computer tomography and electroencephalographic study. Transcranial micropolarization exerts a cerebroprotective effect and has a selective-systemic character due to an increase of neuronal structures activity boht directly in the area of the impact that manifests with the absence of brain edema and the reduction of the destruction locus by 10-15% just after three procedures and as in the other brain regions that results in the decrease of intensity of general cerebral symptoms. The micropolarization promotes restoration of the broken functional connections in central regulatory systems caused by improvement of interaction between neurons, structures and systems the results finally in restoration of central regulation of body's functions. PMID- 17117672 TI - [Use of katadolon in the treatment of abuse headache]. AB - Katadolon (flupirtine), an analgetic with neuroprotective and myorelaxing effect, has been used in the treatment of 16 patients (15 women, 1 man, mean age 38 +/- 6 years) with abuse headache. The diagnosis has been established in accordance with International Classification of Headache Disorders, 2nd Revision (2003). All the patients had episodic headache of tension in anamnesis. Chronic pain and significant tension of pericranial muscles have been registered before the treatment. Patients abused complex analgesics, mean amount of which was 24.5 +/- 6.1 during the period of 21.25 +/- 4 days per month. Monotherapy with katadolon was administered in dosage 100 mg 3 times a day during 1 month. The treatment resulted in significant reduction of frequency, duration and intensity of headache, with a 2-fold decrease in analgesics used. The tension of pericranial muscles decreased and threshold of pain as well as that of nociceptive flexor reflex increased. There was improvement of the patients' quality of life. The period of withdrawal from drug abuse was painless. An effect of treatment remained during 1 month after the withdrawal from the drug in 11 patients, 2 patients continued to receive katadolon. PMID- 17117673 TI - [Experience in clinical use of tanakan in the treatment of the syndrome of mild cognitive impairment]. AB - Clinical efficacy and safety of tanakan has been studied in 30 patients with the syndrome of mild cognitive impairment. The results suggest a clear therapeutic effect of the 6 month course therapy. The efficacy of tanakan was confirmed by significant improvement of cognitive functioning of patients measured with MMSE, MDRS, FAST and other scales. The feature of tanakan effect is its positive influence on memory and other cognitive functions that allows the use of the drug in the treatment of elderly people with the syndrome of mild cognitive impairment. The safety of tanakan used in high therapeutic dosages (240 mg daily) and its good tolerability during chronic administration are revealed. PMID- 17117674 TI - [Anabolic-catabolic balance in depression: an effect of coaxil]. AB - Correlation between anabolic and catabolic process in terms of neuroendocrine changes in depression has been studied before and after coaxil therapy. The index of catabolic processes was cortisol blood level and that of anabolic processes- dehydroepiandrosterone (dehydroepiandrosterone sulfate, DHEA-S) level. The total of 39 patients of middle age were studied: 25 of the study group treated with coaxil (37.5 mg/day during 4 weeks) and 14 of the comparison group treated with sertraline (50 mg/day during 4 weeks). A mean level of cortisol was higher than normal one in patients with depression. Coaxil and sertraline decreased the mean cortisol level, no significant differences being found between these drugs. There was negative correlation between the DHEA-S level and severity of depression before the treatment. Unlike sertraline, coaxil caused an increase of this parameter in patients with decreased DHEA-S level. The ratio cortisol/ DHEA-S decreased during the treatment with either coaxil or sertraline but in the former case it was more pronounced (p = 0.003). The authors considered the data obtained in the aspect of concept of allostasis--the ability of the organism to achieve changes (stress reaction). PMID- 17117675 TI - [Clinical and immunological assessment of efficacy of mexidol in the treatment of lumbosacral radiculopathy]. AB - Efficacy of mexidol in the complex treatment of lumbosacral radiculopathy has been analyzed. Comparing to the traditional treatment using clinical and immunological methods, this method proved to be superior. PMID- 17117676 TI - [Clinical, genealogical and molecular genetic study of Emery-Dreifuss muscular dystrophy]. AB - A search for emerin and lamin A/C (LMNA) mutations was performed in a group of 63 unrelated patients with probable Emery-Dreifuss muscular dystrophy (EDMD) and other MD's with concomitant dilated cardiomyopathy (DCMP). Four different emerin mutations and 7 LMNA mutations were found in unrelated patients. One emerin mutation and 2 LMNA mutations, one of the latter being found twice, have been registered earlier; the rest of the mutations are novel. All the patients with emerin mutations and 3 patients with LMNA mutations represented single cases while 4 LMNA-related cases were familial. De novo origin was proved for one emerin and 3 LMNA mutations. Apart from EDMD phenotypes, varying also in age at onset and severity, 2 cases of limb girdle MD type 1B were diagnosed. One patient with LMNA mutation and severe DCMP had subclinical signs of skeletal myopathy only. There was an overlap between DCMP type 1A and MD's. Autosomal dominant EDMD seems to be more common than "classic" X-linked EDMD. We found neither emerin nor LMNA mutations in a subset of families with EDMD-like phenotypes that may imply an existence of other genes causing similar disorders. Taking into account clinical variability of MD's caused by emerin and LMNA mutations, DNA diagnosis should not confine to the "classic" phenotype. DNA diagnosis of EDMD is important boht for medical genetic counseling and for patients' management: timely diagnosis of the disease allows one to prevent fatal cardiologic complications. PMID- 17117677 TI - [The use of atarax in the treatment of tic hyperkinesia in children: a pilot study]. PMID- 17117678 TI - [Arterial hypertension and the brain]. PMID- 17117679 TI - [Activation of innate immunity in schizophrenia]. PMID- 17117680 TI - Vicinal tricarbonyls and cyano analogs as electrophilic participants in forming pharmacophoric templates for drug discovery. AB - During recent years, fresh attention has been given to a functional group aggregate in which three carbonyl groups have been assembled in a 1,2,3-vicinal relationship. This vicinal tricarbonyl system represents a potent electrophilic unit, usually stabilized in the form of a monohydrate, which, in solution, is in equilibrium with the parent tricarbonyl. It reacts with a range of nucleophiles to form products that are not readily accessible via conventional routes. When an ancillary electrophilic functional group is appended to the vicinal tricarbonyl unit the resulting aggregate serves as a novel di- or trielectrophile in reaction with nucleophiles. This behavior is illustrated by the reaction of vicinal tricarbonyls, attached to vinyl and acetylenic groups, with a broad range of donor reagents to form key components of structures having biological relevance. Substitution of a cyano group for one of the terminal carbonyls leads to labile alpha, beta-diketo nitriles, which can be used as intermediates in forming alpha ketoamide libraries of timely interest as protease inhibitors. PMID- 17117681 TI - Enriching chemical space with diversity-oriented synthesis. AB - The search for new molecular entities in drug discovery and chemical genetic programs requires the screening of high-quality collections of structurally diverse small molecules. The design and synthesis of such collections remains a major challenge to synthetic chemists. Recent strategies and results are presented in the context of the chemical space being interrogated. PMID- 17117682 TI - Automated synthesis of heterocycles on solid supports. AB - The creation of novel diverse heterocycle libraries is an indispensable requirement of modern drug discovery processes. Currently, library sizes of over 10,000 discrete compounds are viable using programmed synthesis on solid supports. This review discusses the recent advances in the automated solid-phase syntheses of heterocycles to generate libraries of bioactive products, and illustrates library sizes that have been obtained, robots used for production of libraries, points of diversity and number of steps on the resin. PMID- 17117683 TI - Compound scale-up at the discovery-development interface. AB - As a result of an economically challenging environment within the pharmaceutical industry, pharmaceutical companies and their departments must increase productivity and cut costs to stay in line with the market. Discovery-led departments such as the medicinal chemistry and lead optimization groups focus on synthesizing large varieties of compounds in minimal amounts, while the chemical development groups must then deliver a few chosen leads employing an optimized synthesis method and using multi-kilogram quantities of material. A research group at the discovery-development interface has the task of medium-scale synthesis which is important in the lead selection stage. The primary objective of this group is the initial scale-up of promising leads for extensive physicochemical and biological testing. The challenge of the interface group involves overcoming synthetic issues within the rigid, accelerated timelines. PMID- 17117684 TI - Biocatalysis: synthesis of chiral intermediates for drugs. AB - Chirality is a key factor in the safety and efficacy of many drug products and thus the production of single enantiomers of drug intermediates has become increasingly important in the pharmaceutical industry. Chiral intermediates and fine chemicals are in high demand for the bulk preparation of drug substances and agricultural products. There has been an increasing awareness of the enormous potential of the use of microorganisms and microorganism-derived enzymes for the transformation of synthetic chemicals with high chemo-, regio- and enantioselectivities. In this article, biocatalytic processes are described for the synthesis of chiral intermediates for drugs. PMID- 17117685 TI - Emerging methods in amide- and peptide-bond formation. AB - Amide bonds are the key chemical connections in proteins and peptides and have a long-established presence in small-molecule therapeutics. Despite their recognized importance, amide bond synthesis relies almost solely on variants of a single reaction: the dehydrative coupling of carboxylic acids and amines. While phenomenally successful in many applications, this process requires superstoichiometric quantities of expensive reagents and precludes chemoselective ligation reactions. This review summarizes emerging chemical alternatives to amide-bond formation with particular emphasis on the development of ligation reactions that permit the coupling of unprotected peptide fragments. PMID- 17117686 TI - Progress toward the discovery and development of efficacious BACE inhibitors. AB - BACE (beta-site amyloid precursor protein [APP] cleavage enzyme) is a transmembrane aspartyl protease responsible for the first cleavage event in the processing of APP to Abeta peptide. Amyloid plaques composed of Abeta peptides are hypothesized to be the root cause of neuronal cell death in Alzheimer's disease patients. Thus, BACE has become a target of significant interest for pharmaceutical and academic research. The recent literature relating to the discovery and development of efficacious BACE inhibitors is reviewed with particular emphasis on the patent literature. PMID- 17117687 TI - Synthesis development of a naphthyridinone p38 kinase inhibitor. AB - In this review the development of a viable large-scale synthesis of a p38 kinase inhibitor is discussed. Multiple strategies have been explored in devising syntheses to the intermediates containing the p38 kinase inhibitor's naphthyridinone core to allow the appendage of difluorophenyl and 4-N-tert butylpiperidine fragments. A novel Heck lactamization reaction was discovered upon reacting 2,6-dichloroacrylanilide with a trihalo-substituted pyridine leading to the rapid synthesis of the naphthyridinone core. Investigations led to the development of two syntheses of 4-N-tert-butyl-chloropiperidine, including a novel methyl Grignard addition to an acetone iminium intermediate to build the tert-butyl group. The chemoselective addition of a 4-N-tertbutyl-chloropiperidine Grignard reagent to a pyridine oxide intermediate followed by re-aromatization using isobutylchloroformate and pyridine as solvent completed the synthesis of this potentially important p38 kinase inhibitor. PMID- 17117688 TI - Achieving polymorph selectivity in the crystallization of pharmaceutical solids: basic considerations and recent advances. AB - Controlling the appearance of crystal polymorphs of pharmaceuticals is essential in developing robust large-scale crystallization processes in the pharmaceutical industry. This review describes many of the thermodynamic and kinetic factors that are important for controlling crystal polymorphism and discusses recent advances in the understanding of the molecular mechanisms of polymorph appearance. Case studies in the development of crystallization processes of pharmaceutical solids, where process analytical technology was applied to resolve the complexities introduced by crystal polymorphism, are discussed. PMID- 17117689 TI - [Oculographic findings in 145 patients with benign paroxysmal positional vertigo]. AB - OBJECTIVE: Benign paroxysmal positional vertigo (BPPV) can be produced by specific manoeuvres and be studied by electrooculography (EOG). It allows an exhaustive study of features in the positional nystagmus. Although most of the patients with BPPV express typical nystagmus, there exists a group of them that exhibit a non typical form. In this work, a comparative study was carried out between two groups of patients. MATERIAL AND METHODS: We have studied retrospectively 145 patients with BPPV and Dix-Hallpicke (D-H) positive manoeuvre registered by EOG techniques. All patients were explored in a complete EOG tests. Two groups were identified: typical response/atypical response according to the features of nystagmus response. Possible alterations in other EOG tests in each group were investigated and differences between both types of response were analyzed. RESULTS: 83.5% of cases showed typical nystagmus and atypical in the rest. Other EOG alterations observed in these patients, were (group typical response/atypical response): caloric hypofunction (17.3%/23%), positional nystagmus (12.4%/62.5%), alterations in visual-oculomotor function tests (10%/50%). CONCLUSIONS: Frequent alterations were observed in the classic manoeuvres and also in the rest of EOG tests. Probably atypical response in the first is due to or can explain the existence of the second. PMID- 17117690 TI - [Clinical tumoral size dissociation in acoustic neuroma: reality or measure distortion?]. AB - INTRODUCTION: In this study we have analyzed (i) the audiometric frequencies more often affected in acoustic neuroma (AN), (ii) the percentage of patients presenting normal hearing and those with sudden hearing loss, (iii) if there is a correlation between tumor size and hearing loss, and (iv) the relationship between clinical and radiological parameters and audiological data. METHODS AND MATERIAL: Retrospective study of 81 patients undergoing surgical removal of a sporadic AN. RESULTS: The highest threshold in the tumor's ear was found at 8000 Hz, and the highest interaural difference at 4000 Hz. The percentage of patients presenting normal hearing and sudden hearing loss was 2.5% and 9%, respectively. No significant association was found between tumor size and hearing loss, preoperative facial palsy or Vth cranial nerve deficit. There was a significant association between the degree of hearing loss and Vth cranial nerve deficit, and between hearing loss and preoperative facial palsy. CONCLUSIONS: The association between hearing loss and Vth cranial nerve deficit, and between hearing loss and preoperative facial palsy is independent the size of the tumour. PMID- 17117691 TI - [Lateral crural "J" flap repair to correct nasal valve collapse]. AB - INTRODUCTION: Nasal valve collapse is a common cause of nasal airway obstruction. Although many techniques have been devised to treat this problem, the outcomes of most of them have been disappointing. The aim of this study is to report our experience using the lateral crural J-flap repair to correct the nasal valve collapse. This technique is based on a new view of the structural etiology of nasal valve collapse and it offers excellent functional and aesthetic outcomes. MATERIAL AND METHODS: We present a retrospective study of 19 patients diagnosed of nasal valve collapse that underwent lateral crural J-flap repair from 2003 to 2005. RESULTS: 28 lateral crural J-flap repairs were made. Six patients underwent concomitant nasal surgery. Mean follow-up was 428 days All the patients have reported improvement of the subjective sensation of inspiratory collapse and there has been no noticeable decrement in benefit over time. No complications due to the procedure were observed. CONCLUSION: The lateral crural J-flap repair is an efective treatment for nasal valve collapse with minimal morbidity and a high success rate. PMID- 17117692 TI - [Results the application of mitomicyn during endonasal and endocanalicular dacryocystorhinostomy by diode laser]. AB - AIM OF THE STUDY: To study the effects, secondary effects and security of the intrasurgical application of mitomicyn C during endonasal and endocanalicular dacryocystorhinostomy with diode laser (TLA-ELA DCR). METHODS: We carried out a randomized, prospective, interventional and double blind study in 200 patients: intrasurgical mitomicyn C was applied in 150 of then (0.4 mgr/0.2 ml) for 5 minutes by means of a polivinil acetate dressing over the osteotomy. In other 50 patients MMC was not used we followed up at 24 hours, 10 days and 1, 3 and 6 months after surgery. Endoscopic aspects and possible complications were valued. The results were compared using the Chi-squared test with the Yates correction. We looked for the presence or abscense of secondary effects in the application of mitomicyn C. RESULTS: The average follow up was 15.25 months (range 6 to 21 months). The percentages of alterations for excesive scarring were 21.77% in patients without and MAC 8.03% in the ones MMC. The difference was statisticaly significant (p = 0.02). We did not find secondary effects due to application of mitomcyn. CONCLUSIONS: Intrasurgical application of topical MMC during TLA-ENL DCR reduces the number of pathological findings due to scarring of the surrouding area of the new drainage without secondary effects. PMID- 17117693 TI - [Epidemiologic analysis of 72 carcinomas of the nasal cavity and paranasal sinuses]. AB - OBJECTIVE: The aim of this study is to define the epidemiological aspects of carcinoma of the nasal cavity and paranasal sinuses. MATERIAL AND METHODS: We performed a retrospective study of 72 carcinomas of the nasal cavity and paranasal sinuses. Various sites, age and sex distribution, drug consumption, TNM stage grouping and treatment were reported. RESULTS: The average age was 63. Seventy- five percent of patients (54/72) were male and 25% (18/72) female. The site of origin was paranasal sinuses in 46 patients (64%), 30 in ethmoid sinus, 15 in maxillary sinus and 1 in sphenoid sinus. Twenty-six patients (36%) were located in nasal cavity. Squamous cell carcinoma was the most frequent histological type in both localizations. The 5-year adjusted survival rate for all patients was 60% (IC: 54-66), 36% (IC: 28-44) for paranasal sinus carcinoma and 86% (IC: 79-93) for nasal cavity carcinoma. The 5-year adjusted survival rate according to the T distribution in 46 carcinomas paranasal sinus was 80% T2, 71% T3, 19% T4a and 6% T4b.(p = 0.0002). CONCLUSIONS: Carcinoma of nasal cavity and paranasal sinuses represent a group of tumors that differ from the rest of carcinomas of the head and neck. PMID- 17117694 TI - [Treatment of early glottic carcinoma with transoral laser CO2]. AB - OBJECTIVE: We present our outcome in the treatment of 125 patients with malignant glottic lesions between 1992 and 2003 in a retrospective study. PATIENT AND METHODS: We analyze 125 cases of patients, with previously untreated carcinoma of the larynx with histological diagnosis, treated with Transoral Laser Microresection. We will pay special attention to those tumors involving the Anterior Commissure. We will also analyze several parameters related to the post and intraoperatory histopathological report, in order to decide adjuvant therapies. We will also analyze the technique indications, complications, hospital length of stay, oncologic and functional outcomes, survival and follow up of recurrences. OUTCOMES AND CONCLUSION: Transoral laser microresection is an effective and functional preserving procedure, and nowadays the base of the treatment of early glottic carcinoma. Anterior Commissure involvement is a prognostic factor. Sometimes complementary therapies are needed based on by the histopathological report. PMID- 17117695 TI - [Distant metastases in head and neck cancer]. AB - INTRODUCTION: The presence of distant metastasis (DM) after the initial treatment of head and neck squamous cell carcinoma (HNSCC) is not considered a common event and it is associated to a poor outcome. PURPOSE: To investigate the prevalence and risk factors associated with the diagnosis of distant metastasis in SCC. METHODS AND MATERIALS: A retrospective study of 633 patients with HNSCC to describe the clinical characteristics of the DM. RESULTS: During the follow-up period after the initial treatment, 6.2% of the patients were diagnosed of having distant metastasis. The site of primary tumor was hypopharynx in 14.4%, unknown origin in 11.8% and oropharynx in 8.5%. The most common sites of DM were the lungs (58%) and the bone (22%). Three year overall survival in patients with DM was 2.5% (versus 49,5% in the control group). CONCLUSIONS: This study confirms that DM have an adverse impact in survival. There is a need of guidelines for screening of distant metastases in patients with HNSCC in order to get an early diagnosis and a more effective treatment. Because of the poor prognosis of DM, protocols including adjuvant chemotherapy should be investigated. PMID- 17117696 TI - [Mechanisms and management of hyperacusis (decreased sound tolerance)]. AB - Hyperacusis is a decreased sound tolerance. Prevalence of the disease is described in 9-15% of the population, but this percentage increases among the tinnitus patients. Pathophysiological mechanisms involve some disruptions in the amplification and regulation processes of the external hair cells or affect the central sound processing at the subcortical level. The role of the serotonin, also involved in other diseases related with hyperacusis (migraine, depression), can be crucial in this disorder. Other theories confirm the effect of the endorphins that activate the excitatory function of the glutamate, the auditory neurotransmitter, increasing its toxicity. The activation of the limbic and autonomic nervous systems produces the emotional reaction of the hyperacusis (anxiety, fear and depression). Proposed treatments are based on acoustic stimulation by a progressive introduction of sound (tinnitus retraining therapy TRT). Noise generators and hearing aids can be fitted in severe cases. The role of some drugs involved in the metabolism of the serotonin open new approaches for the management of hyperacusis. PMID- 17117697 TI - [External auditory canal cholesteatoma as a complication of ear surgery]. AB - External auditory canal cholesteatomas are a rare disease. Their usual clinical appearance is a mass eroding the bony external auditory canal, normally in the inferior or anterior parts, with an intact tympanic membrane and a normal middle ear. A case of this uncommon disease with a review of the scientific literature is presented. Guidelines for the prevention, diagnosis and management are examined. PMID- 17117698 TI - [Rhinolithiasis. Review of one case]. AB - Rhinoliths are foreign bodies of the nose, which may be found during the course of a routine examination or whe the show, if undetected for a long time, symptoms of nasal obstruction, similar to the ones of sinusitis. They require a high level of suspicion to be diagnosed and surgical removal is the treatment of choice. We present the case of a woman diagnosed of rhinolithiasis and we carry out a literature review. PMID- 17117699 TI - My God my choice: the mature minor doctrine and adolescent refusal of life-saving or sustaining medical treatment based upon religious beliefs. PMID- 17117700 TI - The impact of the war over the corporate attorney-client privilege on the business of American health care. PMID- 17117701 TI - An essential prescription: why pharmacist-inclusive conscience clauses are necessary. PMID- 17117702 TI - Legalized importation of Canadian prescription drugs: short-term solution to a long-term problem. PMID- 17117703 TI - The stem cell debate. PMID- 17117704 TI - Biotechnology and the future of humanity. PMID- 17117705 TI - Embryonic human beings. PMID- 17117706 TI - Stem cells: the ethical problems of using embryos for research. PMID- 17117707 TI - Tolerating vs. supporting research that destroys embryos: a difference that can make a moral difference. PMID- 17117708 TI - Altered nuclear transfer: scientific, legal, and ethical foundations. PMID- 17117709 TI - A shared constitutionalism: stem cells and the case for transatlanticism. PMID- 17117710 TI - The transnational perspective of the church: the embryonic cloning debate & stem cell research. PMID- 17117711 TI - Extrapolation of preclinical data into clinical reality translational science. AB - Human and animal in vitro models are potentially powerful preclinical tools: prediction of pharmacological behaviour of drugs; selection of animal species most closely related to humans based on metabolic patterns; prediction of drug interactions and explanation of metabolic origins of interindividual variabilities in pharmacological activity. Extrapolation of preclinical data into clinical reality is a translational science and remains an ultimate challenge in drug development. PMID- 17117712 TI - Smarter candidate selection--utilizing microdosing in exploratory clinical studies. AB - Microdosing offers a faster and potentially less expensive approach to obtaining human in vivo PK data in early clinical drug development. It encompasses the use of pharmacologically inactive doses of test drug in the low microgram range along with ultrasensitive assay methods (PET, AMS) to assess human exposure in order to extrapolate the PK of higher, clinically more relevant doses, assuming linear PK. This strategy allows early evaluation of systemic clearance, oral bioavailability as well as sources of intersubject variability and questions of specific metabolite formation. It does take advantage of reduced regulatory requirements of preclinical safety studies, bulk drug synthesis (CMC requirements) and easier formulation options, e.g., as part of an exploratory IND; however, this is counterbalanced by a need to synthesize radiolabeled test compound and the development of a sophisticated analytical method. Ongoing studies will determine the predictability of human PK using Microdosing methods. PMID- 17117713 TI - The applications of biomarkers in early clinical drug development to improve decision-making processes. AB - Selecting and evaluating biomarkers in drug discovery and early drug development can substantially shorten clinical development time or the time to reach a critical decision point in exploratory drug development. Critical decisions such as candidate selection, early proof of concept/principle, dose ranging, development risks, and patient stratification are based on the appropriate measurements of biomarkers that are biologically and/or clinically validated. The use of biomarkers helps to streamline clinical development by determining whether the drug is reaching and affecting the molecular target in humans, delivering findings that are comparable to preclinical data, and by providing a measurable endpoint that predicts desired or undesired clinical effects and will increase the success rate in the confirmatory stage of clinical development. Appropriateness of biomarkers depends on the stage of development, development strategy, and the nature of the medical indication. Even if a biomarker fails in the validation process there may be still a benefit of having used it. More knowledge about pathophysiology of the disease and the drug has been obtained. Different levels of validation exist at different development phases. Biomarkers are perhaps most useful in the early phase of clinical development when measurement of clinical endpoints may be too time-consuming or cumbersome to provide timely proof of concept or dose-ranging information. Examples of biomarkers are illustrated for the development of new drugs in variant cardiovascular, pulmonary, and CNS diseases. PMID- 17117714 TI - Using exposure-response and biomarkers to streamline early drug development. AB - Biomarkers (BMs) are biological measures of PD drug effects or disease markers that may represent clinically significant patient outcomes, either efficacy or toxicity. Their use in drug development, especially as an integral part of PK/PD modeling, has become a popular strategy for optimizing development time and resources. This approach supports quantitative integration of information across different species and throughout the clinical phases I-III. If the BM is based on the mechanism of action (MOA) of the drug, it is expected to follow an exposure response relationship (E-R). If it is also involved in causal pathways in the pathophysiology of the disease (POD), it may become a surrogate marker (SM). SMs allow prediction of clinical outcomes for different dosing regimens of drug candidates and patient individualization of treatment in clinical practice. Appropriate evaluation of BMs by mechanistic, epidemiological, and clinical pharmacology studies as part of the drug development process allow scientists to establish clinically relevant ER. In early drug development, known ERs for BMs facilitate translation of in vitro findings to in vivo consequences, interspecies PK/PD comparisons, and streamlining of dose-finding phase I and II studies, as well as assessment of new dosing regimen candidates for their likely clinical efficacy and safety, extrapolation of clinical study results to special populations (e.g., pediatrics), and interpretation of exposure differences found in food, drug interaction and special populations studies. Recently, two novel BMs, namely, P50, a measure of ex vivo/in vitro whole blood oxygen affinity and S(pO2), i.e., in vivo pulse oximetry, were used in the development of an allosteric synthetic hemoglobin modifier (SAM), efaproxiral, as PD endpoints; these BMs are based on the MOA of SAMs. Early use of these BMs established excellent in vitro/in vivo PK/PD correlations, appropriate interspecies PK and PD scaling as well as PD-guided phase I and II dose-finding studies. This approach allowed appropriate translation of in vitro and preclinical information along with early identification of sources of PK/PD variability. Frontloading drug development with the identification and use of mechanism-based (MOA/POD) BMs constitutes a rational strategy to quantitatively integrate PK/PD information and optimize dose finding. PMID- 17117715 TI - Experiences with dose finding in patients in early drug development: the use of biomarkers in early decision making. AB - With the increasing cost and complexity of drug development, biomarkers will play an increasing role in the early phases. Biomarkers can be classified into target, mechanistic, or outcome with varying degrees of linkage to disease or treatment effect. They can be used to determine proof of concept by characterising the efficacy or safety profiles, or determining differentiation from any competitor drugs. PK/PD modelling of biomarker data for novel and marketed compounds can be used to predict outpatient dose response. Subsequent simulations may replace or reduce the size and cost of larger phase 2b outpatient studies. Two examples of biomarkers and PK/PD modelling used to characterise dose response are presented. Penile plethysmography (RigiScan Plus) in male erectile dysfunction and phenylephrine challenge urethral pressure in benign prostatic hyperplasia are used to reduce time and cost to reach major exploratory development decision points in these indications. PMID- 17117716 TI - Genotype and phenotype relationship in drug metabolism. AB - Pharmacogenetics, one of the fields of clinical pharmacology, studies how genetic factors influence drug response. If hereditary traits are taken into account appropriately before starting drug treatment, the type of drug and its dosage can be tailored to the individual patient's needs. Today, the relationships between dosage requirements and genetic variations in drug-metabolizing enzymes such as cytochrome P450 (CYP) 2D6, CYP2C9, and CYP2C19 or in drug transporters such as p glycoprotein (ABCB1) and OATP-C (SLC21A6) are substantiated best. A standard dose will bring about more adverse effects than usual if enzymatic activity is lacking or feeble. Sometimes, however, therapeutic response might be better because of higher concentrations: proton pump inhibitors for eradication of Helicobacter pylori are more efficacious in carriers of a deficient CYP2C19 variant. In some cases, genetic tests can help distinguish between responders and nonresponders of a specific drug treatment, and genotype-based dosage is possible. PMID- 17117717 TI - Clinical trials in elderly patients. AB - The increasing size of the elderly population means that both the relative and absolute numbers of prescriptions for elderly patients are increasing. Depending on the age group, between 60% and 80% of elderly people are taking medication, and between 20% and 30% are taking at least three drugs. Prescribing for elderly patients as opposed to younger patients is thus ever more important. This has inevitably meant that the drug development process must increasingly recognise the importance of identifying and developing therapeutic targets relevant to older patients. Clearly, the scientific ethical and regulatory principles that determine conduct of clinical trials in younger individuals apply equally to older people. In addition, the development of drugs to be used in older patients requires an awareness of a number of physiological, pathophysiological and sociological considerations. PMID- 17117718 TI - Dose finding in pediatric patients. AB - It is generally agreed that satisfactory safety and effectiveness of pharmaceutical products for children and adolescents have not yet been established. This applies in particular to anti-cancer drugs and even to those having successfully been used for many years in multidrug chemotherapy protocols for childhood cancer. For example, nephroblastoma or Wilms' tumor is one of the typical and frequent forms of childhood cancer occurring at a median age of about 3 years. Standard therapy for Wilms' tumor is the combination of vincristine and actinomycin D; survival is about 85%. For actinomycin D, the summary of product characteristics states that one contraindication is children aged below 6-12 months. If this would be considered and respected it would mean that a substantial proportion of children with Wilms' tumor would not be treated and thus a proven curative therapy would be withheld. The current situation in pediatrics is that off-label use has become a common practice: in private practice about 20% of prescriptions are off-label, in children's hospitals approximately 40%-50% with 50%-70% in pediatric oncology and more than 90% in neonatology (Conroy et al. 1999, 2000, 2003; Turner et al. 1996, 1998; McIntyre et al. 2000). These conditions are more or less tolerated by the authorities although they are beyond legality. The reason is that appropriate clinical trials like those in adults have not been conducted in children and drugs have therefore not been licensed. PMID- 17117719 TI - Integration of pediatric aspects into the general drug development process. AB - Drug treatment of children is today less regularly based on formal clinical testing than adults. This has led to concerns regarding the safety and efficacy of pediatric medicines and resulted in public action in the United States and the European Union. The reasons for the increasing awareness include better understanding of child physiology, increased trust in GCP (good clinical practice), improved treatment of several severe childhood diseases, a changed view of the child as a subject in society, and more. The US has successfully introduced pediatric legislation that facilitates participation of children in phar maceutical innovation, and comparable approaches are now being discussed in Europe and Japan. While the outcome of the EU pediatric regulation in the near future is still open, the US pediatric legislation has been highly successful over the past 8 years and will be revised before it expires in September 2007. Innovative drugs are today being developed by global pharmaceutical companies. Adding pediatric aspects to this development process is a complex task where companies need to build up internal competency. Bureaucratic procedures that could be harmful to the companies' economic fundaments need to be avoided, and an appropriate ethical framework is required. This needs to be addressed by all partners in healthcare, including regulatory authorities, the pharmaceutical industry, pediatricians, patients and others in a sense of shared responsibility. PMID- 17117720 TI - Current stumbling blocks in oncology drug development. AB - The prognosis of patients with metastatic cancer remains poor and treatment strategies including newer generations of chemotherapeutics have not significantly improved survival in most solid tumors. New approaches are required to further improve patient outcome and survival. Recently, a major leap in the understanding of the molecular mechanisms involved in signal transduction pathways that contribute to tumor growth have been identified as therapeutic targets. Particularly molecules involved in cellular proliferation (e.g., tyrosine kinases) and angiogenesis have been considered as targets for new treatment approaches. Novel therapeutics that specifically target kinase transduction pathways have shown promise as single agents and in combination with standard chemotherapy. In addition, results of recent studies with antiangiogenic monoclonal antibodies validate the use of this class of targeted therapeutics as an important new treatment modality in cancer. This review will focus on the drug development stumbling blocks of targeted treatment modalities in cancer. PMID- 17117721 TI - Exploratory IND: a new regulatory strategy for early clinical drug development in the United States. AB - Optimizing exploratory drug development by means of doing first-in-human studies earlier is an attractive option for pharmaceutical sponsors to select more successful drug candidates. Traditional registration-driven clinical phase 1 programs could be preceded by early human screening studies with subpharmacological single doses (microdoses) or with low pharmacologically active doses of one or several lead candidates, whereby human pharmacokinetic and pharmacodynamic data are generated very early. Such low doses are not expected to have clinically significant toxic potential, so early human screening studies may be supported by abbreviated nonclinical safety packages. Recent U.S. FDA draft guidance (April 2005) regulated early human screening studies conducted under the exploratory IND. The author outlines the features of the study design, dose selection, and nonclinical safety packages required in support of exploratory IND studies in humans. In appropriately chosen cases, exploratory IND could allow for patients' quicker access to safer and more efficacious doses of novel drugs, reduce attrition in clinical trials, and facilitate more economical drug development. PMID- 17117722 TI - Ethnic aspects of cancer trials in Asia. AB - New drugs which have potential in cancer therapy are emerging every day and there is an increasing demand for trial patients all over the world. Asia being the most populated continent, as well as a large market for drug sales, has its own epidemiology of disease. The Asian races also have specific genomics that might affect response to drug treatment. In addition, there are cultural issues to be considered when considering clinical trials. In conclusion, more clinical trials should be done among Asian populations for the best results of drug treatment. PMID- 17117723 TI - Evaluation of the effect on cardiac repolarization (QTc interval) of oncologic drugs. AB - The 12-lead electrocardiograph (ECG) is the standard safety measurement used in clinical trials to identify drug-induced cardiac adverse effects. Drug-induced prolongation of the QTc interval (the measure of cardiac repolarization change), when excessive and in conjunction with the right risk factors, can degenerate into a polymorphic ventricular tachycardia called torsades de pointes and has become a new focus for new drug development. The assessment of an ECG in clinical practice using machine-defined QTc duration is intrinsically unreliable. Current regulatory concepts have focused on the need for measuring ECG intervals using manual techniques using digital processing in a central ECG laboratory. The QT interval is subject to a large degree of spontaneous variability requiring attention to basic clinical trial design issues such as sample size (use as large a cohort as possible), frequency of measurements taken (at least three to six ECGs at baseline and at many time points on therapy with pharmacokinetic samples if possible), and their accuracy. Since most oncologic products are cytotoxic, a Thorough or Dedicated ECG Trial cannot be conducted and in the usual trail, especially in phase I, all changes seen on the ECG will be attributed to the new oncology drug. For most nononcologic drugs, there is regulatory guidance on how much an effect on QTc duration might be related to the risk of cardiac toxicity. For oncology products, the central tendency magnitude and proportion of outliers needs to be well defined to construct a label if the risk-benefit analysis leads to marketing approval. Clinical cardiac findings such as syncope, ventricular tachyarrhythmias, and other cardiac effects will be important in this analysis. PMID- 17117724 TI - The role of PET scanning in determining pharmacoselective doses in oncology drug development. AB - Molecular imaging is the most sensitive and specific method for measuring in vivo molecular pathways in man. Its use in oncology has developed significantly over the last 5-10 years. Molecules can be labelled with positron emitting isotopes and the emitted radiation is detected using sensitive positron emission tomography (PET) cameras. It is now possible to measure in vivo and normal tissue pharmacokinetics of anti-cancer drugs and investigate their mechanism of action. Radiolabelling of tracers can be used to measure specific pharmacodynamic endpoints and target identification. Increasing evidence shows how these technologies, when added to early drug development, can rapidly reduce the time for entry into man and early identification of mechanisms of action. With the move towards more segmented markets and identification of specific subgroups, PET's use for noninvasive biomarkers will become in- creasingly important. However, much international effort between academia and industry is required with prioritisation of development of this technology. PMID- 17117725 TI - Biometrical aspects of drug development. AB - Once the activity of a compound has been established in the laboratory (usually by use of experimental animals) the next stage of development is to bring this forward to humans in early-phase clinical trials. A pharmacokinetic study aims to establish an effective dosing regimen for the compound in order to reach concentrations within the therapeutic window as quickly as possible. The aim of the phase I trials is typically to determine a maximal safe dose with which more rigorous investigation of activity in a phase II trial can be conducted. This chapter deals with statistical issues related to the design of phase I studies. PMID- 17117726 TI - Preventing postmarketing changes in recommended doses and marketing withdrawals. AB - Recent market withdrawals of prescription drug products have brought attention to premarketing safety research. Less known but related to some drug withdrawals are postmarketing dosage changes of newly marketed drugs, including both dosage reductions and increases. These events have serious effects on patients, manufacturers, and regulatory authorities. Most of these harmful events could be avoided by intensive employment of targeted clinical pharmacology investigations to optimize dosage prior to phase III testing and regulatory approval. In this paper, the frequency and implications of postmarketing dosing changes and market withdrawals are considered in light of approaches to preventing them. PMID- 17117735 TI - Ozone artifacts and carbonyl measurements using Tenax GR, Tenax TA, Carbopack B, and Carbopack X adsorbents. AB - Four popular thermally desorbable adsorbents used for air sampling (Tenax TA, Tenax GR, Carbopack B, and Carbopack X) are examined for the potential to form artifacts with ozone (O3) at environmental concentrations. The performance of these adsorbents for the ketone and aldehyde species identified as O3-adsorbent artifacts was also characterized, including recovery, linearity, and method detection limits (MDLs). Using gas chromatography/mass spectrometry, 13 different artifacts were identified and confirmed for both Tenax TA and Tenax GR, 9 for Carbopack B, but none for Carbopack X. Several O3 artifacts not reported previously were identified, including: pentanal, 3-hexanone, 2-hexanone, hexanal, 3-heptanone, and heptanal with Tenax TA; pentanal, 3-hexanone, 2-hexanone, hexanal, and 3-heptanone on Tenax GR; and 1-octene and 1-nonene with Carbopack B. Levels of straight-chain aldehyde artifacts rapidly diminished after a few cycles of adsorbent conditioning/O3 exposure, and concentrations could be predicted using a first-order model. Phenyl-substituted carbonyl artifacts (benzaldehyde and acetophenone) persisted on Tenax TA and GR even after 10 O3 exposure conditioning cycles. O3 breakthrough through the adsorbent bed was most rapid in adsorbents that yielded the highest levels of artifacts. Overall, artifact composition and concentration are shown to depend on O3 concentration and dose, conditioning method, and adsorbent type and age. Calibrations showed good linearity, and most compounds had reasonable recoveries, for example, 90 +/- 15% for Tenax TA, 97 +/- 23% for Tenax GR, 101 +/- 24% for Carbopack B, and 79 +/- 25% (91 +/- 9% for n-aldehydes) for Carbopack X. Benzeneacetaldehyde recovery was notably poorer (22-63% across the four adsorbents). MDLs for several compounds were relatively high, up to 5 ng. By accounting for both artifact formation and method performance, this work helps to identify which carbonyl compounds can be measured using thermally desorbable adsorbents and which may be prone to bias because of the formation of O3-adsorbent artifacts. PMID- 17117736 TI - Emission factors of air toxics from semiconductor manufacturing in Korea. AB - The development of local, accurate emission factors is very important for the estimation of reliable national emissions and air quality management. For that, this study is performed for pollutants released to the atmosphere with source specific emission tests from the semiconductor manufacturing industry. The semiconductor manufacturing industry is one of the major sources of air toxics or hazardous air pollutants (HAPs); thus, understanding the emission characteristics of the emission source is a very important factor in the development of a control strategy. However, in Korea, there is a general lack of information available on air emissions from the semiconductor industry. The major emission sources of air toxics examined from the semiconductor manufacturing industry were wet chemical stations, coating applications, gaseous operations, photolithography, and miscellaneous devices in the wafer fabrication and semiconductor packaging processes. In this study, analyses of emission characteristics, and the estimations of emission data and factors for air toxics, such as acids, bases, heavy metals, and volatile organic compounds from the semiconductor manufacturing process have been performed. The concentration of hydrogen chloride from the packaging process was the highest among all of the processes. In addition, the emission factor of total volatile organic compounds (TVOCs) for the packaging process was higher than that of the wafer fabrication process. Emission factors estimated in this study were compared with those of Taiwan for evaluation, and they were found to be of similar level in the case of TVOCs and fluorine compounds. PMID- 17117737 TI - Effects of wind direction on coarse and fine particulate matter concentrations in southeast Kansas. AB - Field data for coarse particulate matter ([PM] PM10) and fine particulate matter (PM2.5) were collected at selected sites in Southeast Kansas from March 1999 to October 2000, using portable MiniVol particulate samplers. The purpose was to assess the influence on air quality of four industrial facilities that burn hazardous waste in the area located in the communities of Chanute, Independence, Fredonia, and Coffeyville. Both spatial and temporal variation were observed in the data. Variation because of sampling site was found to be statistically significant for PM10 but not for PM2.5. PM10 concentrations were typically slightly higher at sites located within the four study communities than at background sites. Sampling sites were located north and south of the four targeted sources to provide upwind and downwind monitoring pairs. No statistically significant differences were found between upwind and downwind samples for either PM10 or PM2.5, indicating that the targeted sources did not contribute significantly to PM concentrations. Wind direction can frequently contribute to temporal variation in air pollutant concentrations and was investigated in this study. Sampling days were divided into four classifications: predominantly south winds, predominantly north winds, calm/variable winds, and winds from other directions. The effect of wind direction was found to be statistically significant for both PM10 and PM2.5. For both size ranges, PM concentrations were typically highest on days with predominantly south winds; days with calm/variable winds generally produced higher concentrations than did those with predominantly north winds or those with winds from "other" directions. The significant effect of wind direction suggests that regional sources may exert a large influence on PM concentrations in the area. PMID- 17117738 TI - Determining gaseous emission factors and driver's particle exposures during traffic congestion by vehicle-following measurement techniques. AB - Vehicle gaseous emissions (NO, CO, CO2, and hydrocarbon [HC]) and driver's particle exposures (particulate matter < 1 microm [PM1], < 2.5 microm [PM2.5], and < 10 microm [PM10]) were measured using a mobile laboratory to follow a wide variety of vehicles during very heavy traffic congestion in Macao, Special Administrative Region, People's Republic of China, an urban area having one of the highest population densities in the world. The measurements were taken with high time resolution so that fluctuations in the emissions can be seen readily during vehicle acceleration, cruising, deceleration, and idling. The tests were conducted in close proximity to the vehicles, with the inlet of a five-gas analyzer mounted on the front bumper of the mobile laboratory, and the distance between the vehicles was usually within several meters. To measure the driver's particle exposures, the inlets of the particle analyzers were mounted at the height of the driver's breathing position in the mobile laboratory, with the driver's window open. A total of 178 and 113 vehicles were followed individually to determine the gaseous emission factor and the driver's particle exposures, respectively, for motorcycle, passenger car, taxi, truck, and bus. The gaseous emission factors were used to model the roadside air quality, and good correlations between the modeled and monitored CO, NO2, and nitrogen oxide (NO(x)) verified the reliability of the experiments. Compared with petrol passenger cars and petrol trucks, diesel taxies and diesel trucks emitted less CO but more NO(x). The impact of urban canyons is shown to cause a significant increase in the PM1 peak. The background concentrations contributed a significant amount of the driver's particle exposures. PMID- 17117739 TI - Development of an empirical model to estimate real-world fine particulate matter emission factors: the traffic air quality model. AB - The purpose of the study was to quantify the impact of traffic conditions, such as free flow and congestion, on local air quality. The Borman Expressway (I 80/94) in Northwest Indiana is considered a test bed for this research because of the high volume of class 9 truck traffic traveling on it, as well as the existing and continuing installation of the Intelligent Transportation System (ITS) to improve traffic management along the highway stretch. An empirical traffic air quality (TAQ) model was developed to estimate the fine particulate matter (PM2.5) emission factors (grams per kilometer) based solely on the measured traffic parameters, namely, average speed, average acceleration, and class 9 truck density. The TAQ model has shown better predictions that matched the measured emission factor values more than the U.S. Environmental Protection Agency (EPA) PART5 model. During congestion (defined as flow-speeds < 50 km/hr [30 mi/hr]), the TAQ model, on average, overpredicted the measured values only by a factor of 1.2, in comparison to a fourfold underprediction using the EPA-PART5 model. On the other hand, during free flow (defined as flow-speeds > 80 km/hr [50 mi/hr]), the TAQ model was conservative in that it overpredicted the measured values by 1.5-fold. PMID- 17117740 TI - Determination of the adsorptive capacity and adsorption isotherm of vapor-phase mercury chloride on powdered activated carbon using thermogravimetric analysis. AB - This study investigated the use of thermogravimetric analysis (TGA) to determine the adsorptive capacity and adsorption isotherm of vapor-phase mercury chloride on powdered activated carbon (PAC). The technique is commonly applied to remove mercury-containing air pollutants from gas streams emitted from municipal solid waste incinerators. An alternative form of powdered activated carbon derived from a pyrolyzed tire char was prepared for use herein. The capacity of waste tire derived PAC to adsorb vapor-phase HgCl2 was successfully measured using a self designed TGA adsorption system. Experimental results showed that the maximum adsorptive capacities of HgCl2 were 1.75, 0.688, and 0.230 mg of HgCl2 per gram of powdered activated carbon derived from carbon black at 30, 70, and 150 degrees C for 500 microg/m3 of HgCl2, respectively. Four adsorption isotherms obtained using the Langmuir, Freundlich, Redlich-Peterson, and Brunauer-Emmett-Teller (BET) models were used to simulate the adsorption of HgCl2. The comparison of experimental data associated with the four adsorption isotherms indicated that BET fit the experimental results better than did the other isotherms at 30 degrees C, whereas the Freundlich isotherm fit the experimental results better at 70 and 150 degrees C. Furthermore, the calculations of the parameters associated with Langmuir and Freundlich isotherms revealed that the adsorption of HgCl2 by PAC-derived carbon black favored adsorption at various HgCl2, concentrations and temperatures. PMID- 17117741 TI - The adsorptive capacity of vapor-phase mercury chloride onto powdered activated carbon derived from waste tires. AB - Injection of powdered activated carbon (PAC) upstream of particulate removal devices (such as electrostatic precipitator and baghouses) has been used effectively to remove hazardous air pollutants, particularly mercury-containing pollutants, emitted from combustors and incinerators. Compared with commercial PACs (CPACs), an alternative PAC derived from waste tires (WPAC) was prepared for this study. The equilibrium adsorptive capacity of mercury chloride (HgCl2) vapor onto the WPAC was further evaluated with a self-designed bench-scale adsorption column system. The adsorption temperatures investigated in the adsorption column were controlled at 25 and 150 degrees C. The superficial velocity and residence time of the flow were 0.01 m/sec and 4 sec, respectively. The adsorption column tests were run under nitrogen gas flow. Experimental results showed that WPAC with higher Brunauer-Emmett-Teller (BET) surface area could adsorb more HgCl2 at room temperature. The equilibrium adsorptive capacity of HgCl2 for WPAC measured in this study was 1.49 x 10(-1) mg HgCl2/g PAC at 25 degrees C with an initial HgCI2 concentration of 25 microg/m3. With the increase of adsorption temperature < or = 150 degrees C, the equilibrium adsorptive capacity of HgCl2 for WPAC was decreased to 1.34 x 10(-1) mg HgCl2/g PAC. Furthermore, WPAC with higher sulfur contents could adsorb even more HgCl2 because of the reactions between sulfur and Hg2+ at 150 degrees C. It was demonstrated that the mechanisms for adsorbing HgCl2 onto WPAC were physical adsorption and chemisorption at 25 and 150 degrees C, respectively. Experimental results also indicated that the apparent overall driving force model appeared to have the good correlation with correlation coefficients (r) > 0.998 for HgCl2 adsorption at 25 and 150 degrees C. Moreover, the equilibrium adsorptive capacity of HgCl2 for virgin WPAC was similar to that for CPAC at 25 degrees C, whereas it was slightly higher for sulfurized WPAC than for CPAC at 150 degrees C. PMID- 17117742 TI - Comparison of different packing materials for the biofiltration of air toxics. AB - Four different biofilter packing materials (two porous ceramics, perlite, and open pore polyurethane foam) were compared for the removal of toluene vapors. The focus was on evaluating performance at relatively short gas retention time (13.5 and 27 sec). The reactors were initially operated as biotrickling filters with continuous feeding and trickling of a nutrient solution. After significant plugging of the biotrickling filter beds with biomass was observed, the operation mode was switched to biofiltration with only periodic supply of mineral nutrients. This resulted in stable conditions, which allowed detailed investigations over > 6 months. The reactor packed with cattle bone Porcelite (CBP), a ceramic material containing some macronutrients and micronutrients, exhibited the highest performance. The critical load (i.e., load at which 95% removal occurred) was 29 g m(-3) hr(-1) at a gas retention time of 13.5 sec and 66 g m(-3) hr(-1) at a gas retention time of 27 sec. After the long-term experiment, the packing materials were taken from the reactors and examined. The reactors were divided into three sections, top, middle, and bottom, to determine whether spatial differentiation of biomass occurred. The assays included a double staining technique to count total and live microorganisms and determination of moisture, protein, and dry weight contents. Microbial community analysis was also conducted by denaturing gradient gel electrophoresis. The results showed that most reactors had a significant fraction of inactive biomass. Comparatively, the CBP biofilter held significantly higher densities of active biomass, which may be the reason for the higher toluene removal performance. The analyses suggest that favorable material properties and the nutrients slowly released by the CBP provided better environmental conditions for the process culture. PMID- 17117743 TI - Fundamentals of successful monitoring, reporting, and verification under a cap and-trade program. AB - The U.S. Environmental Protection Agency (EPA) developed and implemented the Acid Rain Program (ARP), and NO(x) Budget Trading Programs (NBTP) using several fundamental monitoring, reporting, and verification (MRV) elements: (1) compliance assurance through incentives and automatic penalties; (2) strong quality assurance (QA); (3) collaborative approach with a petition process; (4) standardized electronic reporting; (5) compliance flexibility for low-emitting sources; (6) complete emissions data record required; (7) centralized administration; (8) level playing field; (9) publicly available data; (10) performance-based approach; and (11) reducing conflicts of interest. Each of these elements is discussed in the context of the authors' experience under two U.S. cap-and-trade programs and their potential application to other cap- and trade programs. The U.S. Office of Management and Budget found that the Acid Rain Program has accounted for the largest quantified human health benefits of any federal regulatory program implemented in the last 10 yr, with annual benefits exceeding costs by > 40 to 1. The authors believe that the elements described in this paper greatly contributed to this success. EPA has used the ARP fundamental elements as a model for other cap-and-trade programs, including the NBTP, which went into effect in 2003, and the recently published Clean Air Interstate Rule and Clean Air Mercury Rule. The authors believe that using these fundamental elements to develop and implement the MRV portion of their cap-and-trade programs has resulted in public confidence in the programs, highly accurate and complete emissions data, and a high compliance rate (> 99% overall). PMID- 17117744 TI - Wood smoke contribution to winter aerosol in Fresno, CA. AB - In an effort to better quantify wintertime particulate matter (PM) and the contribution of wood smoke to air pollution events in Fresno, CA, a field campaign was conducted in winter 2003-2004. Coarse and fine daily PM samples were collected at five locations in Fresno, including residential, urban, and industrial areas. Measurements of collected samples included gravimetric mass determination, organic and elemental carbon analysis, and trace organic compound analysis by gas chromatograph mass spectrometry (GC/MS). The wood smoke tracer levoglucosan was also measured in aqueous aerosol extracts using high-performance anion exchange chromatography coupled with pulsed amperometric detection. Sample preparation and analysis by this technique is much simpler and less expensive than derivatized levoglucosan analysis by GC/MS, permitting analysis of daily PM samples from all five of the measurement locations. Analyses revealed low spatial variability and similar temporal patterns of PM2.5 mass, organic carbon (OC), and levoglucosan. Daily mass concentrations appear to have been strongly influenced by meteorological conditions, including precipitation, wind, and fog events. Fine PM (PM2.5) concentrations are uncommonly low during the study period, reflecting frequent precipitation events. During the first portion of the study, levoglucosan had a strong relationship to the concentrations of PM2.5 and OC. In the later portion of the study, there was a significant reduction in levoglucosan relative to PM2.5 and OC. This may indicate a change in particle removal processes, perhaps because of fog events, which were more common in the latter period. Combined, the emissions from wood smoke, meat cooking, and motor vehicles appear to contribute approximately 65-80% to measured OC, with wood smoke, on average, accounting for approximately 41% of OC and approximately 18% of PM2.5 mass. Two residential sites exhibit somewhat higher contributions of wood smoke to OC than other locations. PMID- 17117745 TI - Particle generation by ultraviolet-laser ablation during surface decontamination. AB - A novel photonic decontamination method was developed for removal of pollutants from material surfaces. Such a method relies on the ability of a high-energy laser beam to ablate materials from a contaminated surface layer, thus producing airborne particles. In this paper, the authors presented the results obtained using a scanning mobility particle sizer (SMPS) system and an aerosol particle sizer (APS). Particles generated by laser ablation from the surfaces of cement, chromium-embedded cement, and alumina were experimentally investigated. Broad particle distributions from nanometer to micrometer in size were measured. For stainless steel, virtually no particle > 500 nm in aerodynamic size was detected. The generated particle number concentrations of all three of the materials were increased as the 266-nm laser fluence (millijoules per square centimeter) increased. Among the three materials tested, cement was found to be the most favorable for particle removal, alumina next, and stainless steel the least. Chromium (dropped in cement) showed almost no effects on particle production. For all of the materials tested except for stainless steel, bimodal size distributions were observed; a smaller mode peaked at approximately 50-70 nm was detected by SMPS and a larger mode (peaked at approximately 0.70-0.85 microm) by APS. Based on transmission electron microscopy observations, the authors concluded that particles in the range of 50-70 nm were aggregates of primary particles, and those of size larger than a few hundred nanometers were produced by different mechanisms, for example, massive object ejection from the material surfaces. PMID- 17117746 TI - Performance of a semi-industrial scale gasification process for the destruction of polychlorinated biphenyls. AB - A semi-industrial scale test was conducted to thermally treat mixtures of spent oil and askarels at a concentration of 50,000 ppm and 100,000 ppm of polychlorinated biphenyls (PCBs) under a reductive atmosphere. In average, the dry-basis composition of the synthesis gas (syngas) obtained from the gasification process was: hydrogen 46%, CO 34%, CO2 18%, and CH4 0.8%. PCBs, polychlorinated dibenzo-p-dioxins, and polychlorinated dibenzofurans (PCDDs/PCDFs) in the gas stream were analyzed by high-resolution gas chromatography (GC)-mass spectrometry. The coplanar PCBs congeners 77, 105, 118, 156/ 157, and 167 were detected in the syngas at concentrations < 2 x 10(-7) mg/m3 (at 298 K, 1 atm, dry basis, 7% O2). The chlorine released in the destruction of the PCBs was transformed to hydrogen chloride and separated from the gas by an alkaline wet scrubber. The concentration of PCBs in the water leaving the scrubber was below the detection limit of 0.002 mg/L, whereas the destruction and removal efficiency was > 99.9999% for both tests conducted. The concentration of PCDDs/PCDFs in the syngas were 8.1 x 10(-6) ng-toxic equivalent (TEQ)/m3 and 7.1 x 10(-6) ng-TEQ/m3 (at 298 K, 1 atm, dry basis, 7% O2) for the tests at 50,000 ppm and 100,000 ppm PCBs, respectively. The only PCDD/F congener detected in the gas was the octachloro-dibenzo-p-dioxin, which has a toxic equivalent factor of 0.001. The results obtained for other pollutants (e.g., metals and particulate matter) meet the maximum allowed emission limits according to Mexican, U.S., and European regulations for the thermal treatment of hazardous waste (excluding CO, which is a major component of the syngas, and total hydrocarbons, which mainly represent the presence of CH4). PMID- 17117747 TI - Sources of fine particulate species in ambient air over lake Champlain Basin, VT. AB - This study is a part of an ongoing investigation of the types and locations of emission sources that contribute fine particulate air contaminants to Underhill, VT. The air quality monitoring data used for this study are from the Interagency Monitoring of Protected Visual Environments network for the period of 2001-2003 for the Underhill site. The main source-receptor modeling techniques used are the positive matrix factorization (PMF) and potential source contribution function (PSCF). This new study is intended as a comparison to a previous study of the 1988-1995 Underhill data that successfully revealed a total of 11 types of emission sources with significant contributions to this rural site. This new study has identified a total of nine sources: nitrate-rich secondary aerosol, wood smoke, East Coast oil combustion, automobile emission, metal working, soil/dust, sulfur-rich aerosol type I, sulfur-rich aerosol type II, and sea salt/road salt. Furthermore, the mass contributions from the PMF identified sources that correspond with sampling days with either good or poor visibility were analyzed to seek possible correlations. It has been shown that sulfur-rich aerosol type I, nitrate aerosol, and automobile emission are the most important contributors to visibility degradation. Soil/dust and sea salt/road salt also have an added effect. PMID- 17117748 TI - Can measurement of the intima-media thickness of the carotid artery improve the diagnostic value of exercise stress tests? AB - OBJECTIVE: The diagnostic value of the exercise electrocardiogram (ECG) for the detection of coronary artery disease is hampered by a high proportion of equivocal results, especially in asymptomatic patients. The intima-media thickness of the carotid artery correlates well with the degree of atherosclerosis, also in other vascular beds, such as the coronary arteries. The aim of this study is to evaluate whether measurements of intima-media thickness can improve the diagnostic value of stress ECG in the detection of coronary artery disease. METHODS: Patients without a history of vascular disease and with equivocal exercise stress ECG results were included. The intima-media thickness was measured with high resolution echography at the posterior wall of the common carotid artery, using an automated edge-tracking method. The diagnosis of coronary artery disease was based on the presence of reversible perfusion defects on exercise MIBI-scintigrams. RESULTS: A total of 90 patients (46 men, 44 women) with a mean age of 53.7 years were included. The MIBI-scintigraphy was positive in 17. There was no difference in mean intima-media thickness between MIBI positive and MIBI negative patients (635.76 +/- 84.56 microm and 643.89 +/- 107.06 microm, respectively, p = 0.8). Using multiple regression analysis, neither intima-media thickness measurements, nor classic risk factors could predict the result of MIBI-scintigraphy. Intima-media thickness was mainly influenced by age and hypertension. CONCLUSIONS: In this study, neither intima media thickness measurements, nor classic risk factors could improve the diagnostic value of stress ECG in the detection of coronary artery disease. In case of equivocal stress results, additional cardiovascular imaging techniques remain recommended. PMID- 17117749 TI - Left atrial function in patients with a high C-reactive protein level and paroxysmal atrial fibrillation. AB - OBJECTIVE: We evaluated left atrial dimensions and function in high C-reactive protein (CRP) patients with paroxysmal atrial fibrillation. BACKGROUND: In patients with increased plasma levels of CRP left atrial dysfunction may enhance the occurrence of arrhythmias. METHODS: Two-dimensional and pulsed Doppler echocardiography, were performed in 20 consecutive patients with high CRP levels and paroxysmal atrial fibrillation (group CRf) and in 20 patients with high CRP levels without this arrhythmia (group CR). Twenty normal subjects (group N) were also investigated. Groups were matched for age and gender. RESULTS: CRP was increased in the CRf (median = 1.03 mg/dl), CR (median = 0.84 mg/dl) and N groups (median = 0.23 mg/dl), (p < 0.001) for all comparisons. The CRf, CR and N groups had similar systolic and diastolic blood pressure, left ventricular mass index, left ventricular ejection fraction, isovolumic relaxation time and peak early and late transmitral Doppler flow velocities. Maximal left atrial volume was greater only in the CRf group (54.4 +/- 6.3 ml) compared with the N group (50.3 +/- 4.9 ml, p < 0.05). Left atrial volume preceding atrial contraction was similar in all groups, p=NS. Left atrial minimal volume decreased from 23.0 +/- 1.8 ml in the CRf group, to 19.8 +/- 1.8 ml in the CR group, p < 0.001 and to 18.1 +/- 2.1 ml in the N group, (p < 0.02). The passive emptying fraction of the CRf and CR groups was comparable to that of normal subjects. The CRf group had a decreased left atrial active emptying fraction (0.25 +/- 0.08) compared with the CR (0.36 +/- 0.09) and N groups (0.39 +/- 0.08), p < 0.001 for both comparisons. The reservoir fraction was decreased only in the CRf group compared to normal subjects (1.37 +/- 0.25 vs. 1.82 +/- 0.43, p < 0.001). CONCLUSIONS: These results suggest that the occurrence of paroxysmal atrial fibrillation in patients with a high CRP level is associated with enlargement of the left atrium, depression of its contractile function and is independent of left ventricular hypertrophy and function. The mechanisms linking these variables remain undefined. PMID- 17117750 TI - Age-related changes in myocardial nerve and collagen networks of the auricle of the right atrium. AB - OBJECTIVE: The objective of this study was to analyse age-related changes in human myocardial nerve plexuses and collagen networks of the auricle of the right atrium in subjects in whom no cardiac diseases or pathology had been diagnosed. METHODS AND RESULTS: Morphometric analysis of acetylcholinesterase (AChE)-stained nerve plexuses and picrosirius-stained cardiac collagen networks from 17 persons of both genders aged 20-94 years was performed using video microscopy and a digital video camera. It was found that with age linear regression of nerve plexuses occurred.Atrial collagen content increases lifelong. CONCLUSION: Aging of human atrial myocardium is accompanied by a decrease of nerve plexuses and an increase in fibrosis. PMID- 17117751 TI - Long-term survival and prognostic implications after coronary artery bypass grafting in Chinese patients with coronary artery disease. AB - OBJECTIVES: This hospital-based study was conducted to determine the survival rates of patients after coronary artery bypass grafting (CABG) surgery and the associated prognostic factors related to all-cause mortality during a 7-year follow-up in Taiwan. METHODS AND RESULTS: Between January 1997 and December 2003, the medical records of 1877 patients who underwent primary, isolated CABG surgery were studied. The Kaplan-Meier method was used to estimate survival. Multiple Cox regression was used to investigate the independence of prognostic factors associated with all-cause mortality. Of the 1877 patients who underwent CABG surgery, 192 expired during the 7-year study period. The overall patient survival rate was 85.96% (95% CI: 83.74-88.16). Using multiple Cox regression analysis, in addition to female gender, older age at surgery, pulmonary oedema, longer ischaemic time, longer cardiopulmonary bypass time, and poorer postoperative left ventricular ejection fraction were significant factors associated with all-cause mortality for both men and women. Associated prognostic factors varied by gender. For men, smoking (RR = 2.82, 95% CI: 1.06-4.16), respiratory failure (RR = 6.88, 95% CI: 3.29-14.40) and cardiogenic shock (RR = 4.04, 95% CI: 2.13-7.67) were significantly related to all-cause mortality, but not for women. Sepsis (RR = 8.97,95% CI: 1.19-19.81) and ICU stay (RR = 1.03,95% CI: 1.01-1.05) were significantly related to all-cause mortality among female patients only. CONCLUSIONS: Several gender-related differences were noted pertaining to all cause mortality and the relationships between smoking, sepsis, respiratory failure, cardiogenic shock, and ICU stay. PMID- 17117752 TI - Uric acid in patients with angiographically documented coronary heart disease. AB - OBJECTIVE: High serum uric acid levels have been associated with coronary heart disease (CHD). Hyperuricaemia is known to be related to several of the established aetiologic risk factors of CHD, such as obesity, insulin resistance, raised serum triglycerides, and hypertension, but it is still uncertain whether this relationship may cause the association between CHD and uric acid. We have investigated whether uric acid was an independent risk factor for CHD. METHODS AND RESULTS: Two-hundred and ninety patients with suspected CHD referred to elective coronary angiography were enrolled. The association between angiographically detected coronary stenosis > 50% in one or more of the 3 major coronary arteries and uric acid levels was examined. A significant association between high uric acid levels and the presence of one or more diseased vessels in women (p = 0.02) was observed, while no association was found in men (p = 0.25). After adjustment for several possible confounders such as smoking, triglycerides, HDL cholesterol, and a diagnosis of diabetes mellitus the association among women disappeared. CONCLUSION: The results indicate that uric acid should not be viewed as an independent risk factor for CHD, but more likely as a biological marker reflecting other causative parameters. PMID- 17117753 TI - Changes in escape rhythms several years after radiofrequency ablation of the atrioventricular junction combined with pacemaker implantation. AB - OBJECTIVE: Escape rhythm is thought to play a considerable role in protection against adverse outcome due to pacemaker malfunction. We studied the escape rhythms in 32 patients with supraventricular tachyarrhythmia refractory to medical therapy who underwent radiofrequency ablation of the atrioventricular junction combined with implantation of a pacemaker. METHODS AND RESULTS: We performed the escape rhythm analysis immediately and 2.6 +/- 1.9 years after the radiofrequency ablation by decreasing the pacing rate. In the initial study, escape rhythms (41 +/- 8 beats/min) were documented in 20 patients (63%). Non cardiac death occurred in 3 patients with escape rhythm, and cardiac death occurred in I patient without escape rhythm. In the follow-up study, escape rhythms were detected in 22 of 28 patients (79%). Escape rhythm had developed in 6 (55%) of 11 patients who were without escape rhythm initially, while escape rhythm disappeared in 1 of 17 (5.9%) patients who had escape rhythm initially. The changes in escape rhythm were not related to QRS width of the intrinsic beat. There was no correlation between the number of radiofrequency applications or the ratio between atrial and ventricular electrocardiogram voltages of radiofrequency applications and the development of escape rhythms. CONCLUSIONS: The present long term follow-up study demonstrated that the emergence of an escape rhythm increased several years after ablation, but was unrelated to procedural factors. There are, however, certain patients in whom the disappearance of escape rhythms occur. The evaluation of escape rhythms is thus necessary to determine the risk associated with pacemaker failure. PMID- 17117754 TI - Late-term results of mitral valve replacement with St. Jude Medical mechanical valve prosthesis: Samsun experience. AB - BACKGROUND AND AIM OF THE STUDY: We have reported the short- and long-term results of mitral valve replacement in this article. METHODS: Mitral valve replacement was conducted in 276 patients in our clinic between January 1989 and March 2005. The youngest patient was 4 years old and the oldest patient was 74 years old. Mean age was 40.08 +/- 1.06 y. Of these patients, 41.3% were men and 58.7% were women. The reason for operation was mitral stenosis in 96 patients (34.78%), mitral insufficiency in 78 patients (29.26%) and mitral stenosis plus mitral insufficiency in 102 patients (36.96%). The aetiology of mitral valve lesions was acute rheumatic fever in 208 patients (75.36%). The aetiology of mitral valve lesions was degenerative in 37 patients (13.41%), ischaemic in 23 patients (8.33%) and congenital in 8 patients (2.9%). RESULTS: In the 5, 10 and 15-year periods, the actual survival rates were 87.64% +/- 2.02%, 83.35% +/- 2.38% and 68.19% +/- 5.63%, respectively. Thromboembolism was observed in 38 patients (13.77%). The rates of actual freedom from thromboembolism in the 5, 10 and 15-year periods were 93.08% +/- 1.53%, 88.48% +/- 1.99% and 81.06% +/- 3.43%, respectively. Of the 276 patients who had been observed for 15 years, 5 had (1.81%) valvular thrombosis. The rates of actual freedom from valvular thrombosis in the 5, 10 and 15-year periods were 98.89% +/- 0.64%, 98.04% +/- 0.87% and 98.04% +/- 0.87%, respectively. In the 15-year period, 23 patients (8.33%) had haemorrhage due to anti-coagulation. The rates of actual freedom from haemorrhage due to anti-coagulation in the 5, 10 and 15-year periods were 95.64% +/- 1.23%, 93.40% +/- 1.56% and 87.73% +/- 2.96%, respectively. Seven patients (2.54%) had prosthetic valvular endocarditis. The rates of actual freedom from endocarditis in the 5, 10 and 15-year periods were 98.51% +/- 0.74%, 97.60% +/- 0.97% and 97.01% +/- 1.13%, respectively. Nine patients (3.27%) were re-operated. The rates of actual freedom from re-operation in the 5, 10 and 15-year periods were 97.45% +/- 0.95%, 96.58% +/- 1.12% and 96.58% +/- 1.12%, respectively. CONCLUSIONS: St. Jude Medical mechanical valve prosthesis has been the valve of choice in our clinic owing to its excellent haemodynamic properties and low rates of complication. PMID- 17117755 TI - Role of myocardial bridging in the apical localization of stress cardiomyopathy. AB - BACKGROUND: The apical ballooning syndrome is precipitated by emotional or physical stress but the underlying mechanism remains poorly understood. The contribution of myocardial bridging on the aetiology and the onset of the syndrome is not known. METHODS: We observed 8 patients with chest pain, T-wave inversion in several leads of the ECG, transient left ventricular apical ballooning and no significant angiographic stenosis. RESULTS: There were 7 women and I man. The median age was 67.5 years. Seven patients had an intense emotional or physical stress (87.5%). All patients presented with chest pain and aT-wave inversion in the precordial leads. The median elevation of creatine-kinase was 171 IU. In all patients, echocardiography showed an alteration of the left ventricular function with a very extensive apical akinesia. Left ventricular hypertrophy was observed in 7 patients. A myocardial bridging in the mid segment of the left anterior descending coronary artery was observed in 5 patients (62.5%). Recovery was complete in all patients. During follow-up, no patient showed recurrence. CONCLUSIONS: Our data suggest that myocardial bridging possibly enhanced by catecholamines during stress may contribute, in association with left ventricular hypertrophy, to the preferential apical localization of the apical ballooning syndrome. Further investigations are necessary to confirm PMID- 17117756 TI - The management of familial hypercholesterolaemia in Morocco. AB - Familial hypercholesterolaemia (FH) is an autosomal dominant disorder characterized by high levels of low-density lipoprotein cholesterol (LDL-C) as a result of mutations that impair their removal from plasma.The clinical consequence is a high risk of premature cardiovascular disease. Because of the extreme risk of mortality and morbidity, diagnosis, recruitment and management of FH patients must be one of the priorities of public health. In Morocco, specialized consultation for dyslipidaemia and strategy for management of this cardiovascular major risk factor does not exist, making FH identification and management difficult. In this review, we present the first FH state of the art in our country through a sample of 66 subjects. By this analysis, we have tried to elucidate some points that impede the identification and recruitment of heterozygous FH and the management of both heterozygous and homozygous FH in Morocco. Also, we have attempted to propose some strategies for an adequate management of FH in our country, taking into account the local specifications. PMID- 17117757 TI - Histopathology of calcific aortic valve stenosis. AB - Calcific aortic valve stenosis is the most common and increasing heart valve disease in the western world. In the last 30 years, diagnosis and management were revolutionized by the development of cardiac catheterisation, echocardiography, cardiac surgery, and medication. Recently, new strategies were introduced for aortic valve replacement using more sophisticated bioprosthetic heart valves. Moreover, tissue-engineered heart valves are under development to improve management strategies. In this article we review the current morphological and histopathological findings in the progression of calcific aortic valve stenosis. This is, to our understanding, important to contribute to the knowledge of fundamental management strategies of this disease. PMID- 17117758 TI - Amiodarone and polymorphic ventricular tachycardia: to slow down or to accelerate the underlying rhythm? AB - Polymorphic ventricular tachycardia (VT) comprises a variety of different subforms. Two patients were admitted to our hospital and developed polymorphic VT under treatment with amiodarone. However, each of them turned out to have a very different form of the arrhythmia and therefore required a different therapeutic approach. The clinical decision-making required a distinction between the two forms, and the following therapeutic approach included terminating the administration of amiodarone in the first case and continuing administration in the other case. As the most common forms of polymorphic VT are described, it becomes clear that each different form should be identified because of the important therapeutic implications. In particular, the role of amiodarone can be quite different in the management of these different forms of polymorphic VT. PMID- 17117759 TI - Non-invasive imaging of a giant right coronary artery due to a coronary fistula. AB - Coronary artery fistula is a relatively rare finding in patients undergoing diagnostic cardiac catheterization. Incidence, angiographic characteristics and natural history of coronary fistulas of this type have not been well defined in the literature. We report a case of a 54-year-old man in whom such abnormalities have been diagnosed with invasive and non-invasive coronary imaging and review the epidemiology, treatment and prognosis of coronary fistulas. PMID- 17117760 TI - Frequent ventricular premature beats: a reversible cause of cardiomyopathy? PMID- 17117761 TI - Tirofiban-associated acute thrombocytopenia. AB - Randomized clinical trials of glycoprotein IIb/IIIa inhibition during percutaneous coronary intervention have shown significant clinical efficacy and safety in reducing the morbidity and mortality. In Turkey, only tirofiban is available. Tirofiban is a safe and effective agent in combination with heparin and aspirin in the setting of an acute coronary syndrome. In previous studies involving tirofiban, the incidence of bleeding complications was more frequent than heparin alone but major bleeding complications were seen in 5.3% of patients receiving tirofiban and not significantly different from the heparin-only treated patients.The incidence of thrombocytopenia, defined as an absolute platelet count of < 90,000/mm3, was 0.4% in the PRISM, 1.9% in PRISM-PLUS and 1.1% in RESTORE trials. We reported a case of acute profound thrombocytopenia 48 hours after the administration of tirofiban in the treatment of a patient with an acute coronary syndrome and tirofiban discontinuation was sufficient in the management of this condition. PMID- 17117762 TI - Effect of contact pressure on wear and friction of ultra-high molecular weight polyethylene in multidirectional sliding. AB - Computational wear models need input data from valid tribological tests. For the wear model of a total hip prosthesis, the contact pressure dependence of wear and friction of ultra-high molecular weight polyethylene (UHMWPE) against polished CoCr in diluted calf serum lubricant was studied, and useful input data produced. Two test devices were designed and built: a heavy load circularly translating pin on-disc (HL-CTPOD) wear test device and an HL-CTPOD friction measurement device. Both can be used with a wide range of loads. The wear surface diameter of the test pin was kept constant at 9 mm, whereas the load was varied so that the nominal contact pressure ranged from 0.1 to 20 MPa. The wear factor decreased with increasing contact pressure, whereas the coefficient of friction first increased with increasing contact pressure with low pressure values and then decreased. Up to the pressure of 2.0 MPa, the wear mechanisms and wear factors were in good agreement with clinical findings. In the critical range of 2.0-3.5 MPa, the wear mechanisms and wear factors started to differ from clinical ones, and the decrease of the wear factor steepened. The discrepancy became more and more evident as the pressure was gradually increased beyond 3.5 MPa. It appears that the pressure value of 2.0 MPa should not be exceeded in pin-on-disc wear tests that are to reproduce the clinical wear of UHMWPE acetabular cups. PMID- 17117763 TI - Effect of ultra-high molecular weight polyethylene thickness on contact mechanics in total knee replacement. AB - One of the important design parameters in current knee joint replacements is the thickness of the ultra-high molecular weight polyethylene (UHMWPE) tibial insert, yet there is no clear definition of the upper limit of the 'thick' polyethylene insert. Using one design knee implant and subjecting it to the physiological loads encountered throughout the gait cycle, measurements of the lengths of contact imprints generated were compared with the corresponding theoretical predictions for different insert thicknesses under the same applied load. Multiple regression analysis was applied to test whether the dimensions of contact imprints are influenced by UHMWPE thickness. Good agreement was obtained between the theoretical predictions and the experimental measurements of the dimensions of contact imprints when the knee was at 60 degrees flexion. Therefore, it was possible to estimate the contact pressure at the articulating surface using the theoretical model. Contact imprint dimensions increased with increasing applied load. Statistical analysis of the experimental data revealed that, at 0 degree flexion, the overall imprint dimensions increased as the UHMWPE thickness increased from 8 to 20 mm. However, the increment was not significant when the thickness subinterval 10-15 mm was considered. Furthermore, at 60 degrees flexion, thickness was not a significant factor for the overall imprint dimensions. No evidence was found from the data to suggest that an increment in polyethylene thickness over 10 mm would significantly reduce the contact imprint dimensions. These findings suggest that thicker inserts can be avoided, as they require unnecessary bone resection. PMID- 17117764 TI - The effect of angle on wall shear stresses in a LIMA to LAD anastomosis: numerical modelling of pulsatile flow. AB - The purpose of this study was to examine the effect of anastomotic angle on the flow patterns and wall shear distributions at the distal anastomosis of a left interior mammary artery (LIMA) graft to the left anterior descending artery (LDA). It is now well recognized that abnormal wall shear stress distributions along the anastomotic bed, around the toe, and around the heel can contribute to the focal development of intimal hyperplasia. However, the exact nature of the interaction between the dominant pulsing flow and the anastomotic angle on wall shear stresses has not been fully investigated numerically. In this study a commercial CFD package was used for three-dimensional flow analysis where the pulsatile waveforms and flowrates used as the boundary conditions are representative of an anastomosed left internal mammary artery and a stenosed left anterior descending coronary artery (intermediate, <70 per cent diameter narrowing). The flow patterns and distributions of time-averaged wall shear stress (TAWSS) and the oscillatory shear index (OSI) for three anastomotic angles of 20, 40, and 60 degrees were evaluated and compared with other published data. The findings indicated that transient, highly disturbed flow patterns occurred in localized regions of the proximal and distal native segments and in the anastomotic domain including recirculation zones, moving points of stagnation, and oscillating wall shear stresses mainly on the bed, at the toe, and at the heel. Moreover, higher anastomotic angles resulted in more extreme variations in TAWSS and OSI values, particularly around the toe and along the bed. In addition, the effect of anastomotic angle on OSI values at the heel followed the same pattern whereas the TAWSS values along the graft at the heel showed a significant increase at the lowest anastomotic angle of 20 degrees. PMID- 17117765 TI - Preoperative planning and intraoperative guidance for accurate computer-assisted minimally invasive hip resurfacing surgery. AB - Hip resurfacing is an alternative to total hip replacement (THR) and is particularly suitable for the younger, more active patient. However, it is a more demanding procedure. This paper describes a system that enables the surgeon to plan the surgery preoperatively with optimally sized and placed components, and then transfer this plan to an intraoperative system that registers computer models to the real patient and tracks surgical tools, allowing the surgeon to ensure that the bone is resected correctly and that the components are fitted in accordance with the plan. The paper describes a series of instruments used with the system which are locked to the bone. These instruments serve the dual purpose of soft tissue retraction and bone immobilization. The system will shortly be the subject of laboratory and clinical evaluation. Registration, a cornerstone of the tracked instrument system, has been tested, and accuracy measures are provided. Experimental results for the remainder of the system will be provided after clinical trials. PMID- 17117766 TI - Development of the mennen 3 peripro fixation plate for the treatment of periprosthetic fractures of the femur. AB - The Mennen femur plate is an internal fixation device used for the management of femoral perisprosthetic fractures, usually after total hip replacement surgery. The implant uses a number of curved prongs that embrace the fractured bone around its circumference without interfering with the stem of the prosthesis. Although the device has been used with considerable clinical success since its first introduction, a number of negative clinical results have been reported in the literature. The failure modes of the device are described and an evaluation of its performance is briefly presented. Based on this assessment as well as comments in the open literature, modifications in the design of the device have been implemented. The new Mennen 3 PeriPro plate is presented, with all the necessary data for a coherent explanation of its improved characteristics as defined using numerical simulations and experimental tests. The new device has all the beneficial features of the previous plate with improved structural performance and fatigue life and new sculpted formation of the prongs, providing a simple implantation technique with maximum gripping and minimum effort from the surgeon. The unique mode of fixation has been further improved, providing ample anchorage on the fracture bone without compromising its biomechanical integrity. By combining the device with a cable system, the spectrum of applications will be further expanded, enabling the surgeon to treat a broader range of fracture patterns. PMID- 17117767 TI - Radiopacity of tantalum-loaded acrylic bone cement. AB - Radiopacifying agents are commonly added to bone cements to enhance the visibility of the cement in radiography. The radiopacifiers usually employed may, however, have undesired effects on the mechanical properties of the cement. A potentially new radiopacifier is tantalum, which in the present work was evaluated in terms of radiopacity. Bone cements containing different percentages of tantalum were compared with plain bone cement as well as with formulations containing different percentages of the commonly used radiopacifier barium sulphate. The radiopacity was assessed quantitatively and qualitatively, by measuring with a digital densitometer the optical density of the cement on X-ray films, and consulting the expertise of ten orthopaedic surgeons. It was found that tantalum does present radiopacity, but not as high as barium sulphate under the specific conditions applied to this study. PMID- 17117768 TI - Trial fitting of a removable partial denture framework made using computer-aided design and rapid prototyping techniques. AB - Previous studies of CAD/CAM-produced sacrificial patterns for removable partial denture frameworks have been documented but to date, no such restorations have been test-fitted to a patient. This paper provides details of the first trial fitting to a patient of an RPD framework, the sacrificial pattern of which was produced by CAD/CAM and RP technologies. A cast of the patient was scanned and the normal procedures of dental surveying and pattern build were undertaken with reference to the scanned model using computer-aided design. A sacrificial pattern of the design was produced by rapid prototyping technology. After spruing the pattern, investment-casting and finishing techniques were carried out according to conventional principles. The framework was successfully trial-fitted to the patient and clinically judged to be acceptable for the next stage of denture fabrication, that of adding acrylic bases and artificial teeth. PMID- 17117769 TI - Validation in medical image processing. PMID- 17117770 TI - Twenty new digital brain phantoms for creation of validation image data bases. AB - Simulations provide a way of generating data where ground truth is known, enabling quantitative testing of image processing methods. In this paper, we present the construction of 20 realistic digital brain phantoms that can be used to simulate medical imaging data. The phantoms are made from 20 normal adults to take into account intersubject anatomical variabilities. Each digital brain phantom was created by registering and averaging four T1, T2, and proton density (PD)-weighted magnetic resonance imaging (MRI) scans from each subject. A fuzzy minimum distance classification was used to classify voxel intensities from T1, T2, and PD average volumes into grey-matter, white matter, cerebro-spinal fluid, and fat. Automatically generated mask volumes were required to separate brain from nonbrain structures and ten fuzzy tissue volumes were created: grey matter, white matter, cerebro-spinal fluid, skull, marrow within the bone, dura, fat, tissue around the fat, muscles, and skin/muscles. A fuzzy vessel class was also obtained from the segmentation of the magnetic resonance angiography scan of the subject. These eleven fuzzy volumes that describe the spatial distribution of anatomical tissues define the digital phantom, where voxel intensity is proportional to the fraction of tissue within the voxel. These fuzzy volumes can be used to drive simulators for different modalities including MRI, PET, or SPECT. These phantoms were used to construct 20 simulated T1-weighted MR scans. To evaluate the realism of these simulations, we propose two approaches to compare them to real data acquired with the same acquisition parameters. The first approach consists of comparing the intensities within the segmented classes in both real and simulated data. In the second approach, a whole brain voxel-wise comparison between simulations and real T1-weighted data is performed. The first comparison underlines that segmented classes appear to properly represent the anatomy on average, and that inside these classes, the simulated and real intensity values are quite similar. The second comparison enables the study of the regional variations with no a priori class. The experiments demonstrate that these variations are small when real data are corrected for intensity nonuniformity. PMID- 17117771 TI - Phenomenological model of diffuse global and regional atrophy using finite element methods. AB - The main goal of this work is the generation of ground-truth data for the validation of atrophy measurement techniques, commonly used in the study of neurodegenerative diseases such as dementia. Several techniques have been used to measure atrophy in cross-sectional and longitudinal studies, but it is extremely difficult to compare their performance since they have been applied to different patient populations. Furthermore, assessment of performance based on phantom measurements or simple scaled images overestimates these techniques' ability to capture the complexity of neurodegeneration of the human brain. We propose a method for atrophy simulation in structural magnetic resonance (MR) images based on finite-element methods. The method produces cohorts of brain images with known change that is physically and clinically plausible, providing data for objective evaluation of atrophy measurement techniques. Atrophy is simulated in different tissue compartments or in different neuroanatomical structures with a phenomenological model. This model of diffuse global and regional atrophy is based on volumetric measurements such as the brain or the hippocampus, from patients with known disease and guided by clinical knowledge of the relative pathological involvement of regions and tissues. The consequent biomechanical readjustment of structures is modelled using conventional physics-based techniques based on biomechanical tissue properties and simulating plausible tissue deformations with finite-element methods. A thermoelastic model of tissue deformation is employed, controlling the rate of progression of atrophy by means of a set of thermal coefficients, each one corresponding to a different type of tissue. Tissue characterization is performed by means of the meshing of a labelled brain atlas, creating a reference volumetric mesh that will be introduced to a finite-element solver to create the simulated deformations. Preliminary work on the simulation of acquisition artefacts is also presented. Cross-sectional and longitudinal sets of simulated data are shown and a visual classification protocol has been used by experts to rate real and simulated scans according to their degree of atrophy. Results confirm the potential of the proposed methodology. PMID- 17117772 TI - Creation and application of a simulated database of dynamic [18f]MPPF PET acquisitions incorporating inter-individual anatomical and biological variability. AB - During the process of validation of a new tracer, estimation of performance and validation of processing algorithms have to be investigated with data sets representative of the ground truth. Because this ground truth is hardly accessible in positron emission tomography (PET), validations of processing algorithms often rely on the use of simulated data sets. Considering that Monte Carlo simulators are very time consuming and are not very easy to use, the building of publicly available databases of simulated PET volumes are becoming highly desirable. We present here the methodology employed for the creation of a database of simulated dynamic [18F]MPPF-PET data, including inter-individual anatomical and biological variability which meets the criteria of a gold standard database as defined by Lehmann: reliance, equivalence, independence, relevance, significance. The assessment of the realism of the built database against actual MPPF PET data is also presented here. Whereas the database was specifically created for the investigations of quantification of activity and binding of ligand-receptor with the [18F]MPPF PET tracer, it may serve the community with countless purposes. The full strength of this database, does not only stem from the knowledge of important information such as the true activity map and underlying anatomical data, but also from the possibility to fully control the biological difference between sets of simulated PET data. Indeed, time activity curves included in the simulated data sets are controlled by a multicompartmental model of ligand-receptor exchanges. This latter feature is of a great interest in the context of the improvement of the detectability of biological variation in PET. PMID- 17117773 TI - A cross validation study of deep brain stimulation targeting: from experts to atlas-based, segmentation-based and automatic registration algorithms. AB - Validation of image registration algorithms is a difficult task and open-ended problem, usually application-dependent. In this paper, we focus on deep brain stimulation (DBS) targeting for the treatment of movement disorders like Parkinson's disease and essential tremor. DBS involves implantation of an electrode deep inside the brain to electrically stimulate specific areas shutting down the disease's symptoms. The subthalamic nucleus (STN) has turned out to be the optimal target for this kind of surgery. Unfortunately, the STN is in general not clearly distinguishable in common medical imaging modalities. Usual techniques to infer its location are the use of anatomical atlases and visible surrounding landmarks. Surgeons have to adjust the electrode intraoperatively using electrophysiological recordings and macrostimulation tests. We constructed a ground truth derived from specific patients whose STNs are clearly visible on magnetic resonance (MR) T2-weighted images. A patient is chosen as atlas both for the right and left sides. Then, by registering each patient with the atlas using different methods, several estimations of the STN location are obtained. Two studies are driven using our proposed validation scheme. First, a comparison between different atlas-based and nonrigid registration algorithms with a evaluation of their performance and usability to locate the STN automatically. Second, a study of which visible surrounding structures influence the STN location. The two studies are cross validated between them and against expert's variability. Using this scheme, we evaluated the expert's ability against the estimation error provided by the tested algorithms and we demonstrated that automatic STN targeting is possible and as accurate as the expert-driven techniques currently used. We also show which structures have to be taken into account to accurately estimate the STN location. PMID- 17117774 TI - Generalized overlap measures for evaluation and validation in medical image analysis. AB - Measures of overlap of labelled regions of images, such as the Dice and Tanimoto coefficients, have been extensively used to evaluate image registration and segmentation algorithms. Modern studies can include multiple labels defined on multiple images yet most evaluation schemes report one overlap per labelled region, simply averaged over multiple images. In this paper, common overlap measures are generalized to measure the total overlap of ensembles of labels defined on multiple test images and account for fractional labels using fuzzy set theory. This framework allows a single "figure-of-merit" to be reported which summarises the results of a complex experiment by image pair, by label or overall. A complementary measure of error, the overlap distance, is defined which captures the spatial extent of the nonoverlapping part and is related to the Hausdorff distance computed on grey level images. The generalized overlap measures are validated on synthetic images for which the overlap can be computed analytically and used as similarity measures in nonrigid registration of three dimensional magnetic resonance imaging (MRI) brain images. Finally, a pragmatic segmentation ground truth is constructed by registering a magnetic resonance atlas brain to 20 individual scans, and used with the overlap measures to evaluate publicly available brain segmentation algorithms. PMID- 17117775 TI - Nonrigid 2-D/3-D registration for patient specific bronchoscopy simulation with statistical shape modeling: phantom validation. AB - This paper presents a nonrigid registration two-dimensional/three-dimensional (2 D/3-D) framework and its phantom validation for subject-specific bronchoscope simulation. The method exploits the recent development of five degrees-of-freedom miniaturized catheter tip electromagnetic trackers such that the position and orientation of the bronchoscope can be accurately determined. This allows the effective recovery of unknown camera rotation and airway deformation, which is modelled by an active shape model (ASM). ASM captures the intrinsic variability of the tracheo-bronchial tree during breathing and it is specific to the class of motion it represents. The method reduces the number of parameters that control the deformation, and thus greatly simplifies the optimisation procedure. Subsequently, pq-based registration is performed to recover both the camera pose and parameters of the ASM. Detailed assessment of the algorithm is performed on a deformable airway phantom, with the ground truth data being provided by an additional six degrees-of-freedom electromagnetic (EM) tracker to monitor the level of simulated respiratory motion. PMID- 17117776 TI - Automatic contour propagation in cine cardiac magnetic resonance images. AB - We have developed a method for automatic contour propagation in cine cardiac magnetic resonance images. The method consists of a new active contour model that tries to maintain a constant contour environment by matching gray values in profiles perpendicular to the contour. Consequently, the contours should maintain a constant position with respect to neighboring anatomical structures, such that the resulting contours reflect the preferences of the user. This is particularly important in cine cardiac magnetic resonance images because local image features do not describe the desired contours near the papillary muscle. The accuracy of the propagation result is influenced by several parameters. Because the optimal setting of these parameters is application dependent, we describe how to use full factorial experiments to optimize the parameter setting. We have applied our method to cine cardiac magnetic resonance image sequences from the long axis two chamber view, the long axis four-chamber view, and the short axis view. We performed our optimization procedure for each contour in each view. Next, we performed an extensive clinical validation of our method on 69 short axis data sets and 38 long axis data sets. In the optimal parameter setting, our propagation method proved to be fast, robust, and accurate. The resulting cardiac contours are positioned within the interobserver ranges of manual segmentation. Consequently, the resulting contours can be used to accurately determine physiological parameters such as stroke volume and ejection fraction. PMID- 17117777 TI - Evaluation of four probability distribution models for speckle in clinical cardiac ultrasound images. AB - Segmenting cardiac ultrasound images requires a model for the statistics of speckle in the images. Although the statistics of speckle are well understood for the raw transducer signal, the statistics of speckle in the image are not. This paper evaluates simple empirical models for first-order statistics for the distribution of gray levels in speckle. The models are created by analyzing over 100 images obtained from commercial ultrasound machines in clinical settings. The data in the images suggests a unimodal scalable family of distributions as a plausible model. Four families of distributions (Gamma, Weibull, Normal, and Log normal) are compared with the data using goodness-of-fit and misclassification tests. Attention is devoted to the analysis of artifacts in images and to the choice of goodness-of-fit and misclassification tests. The distribution of parameters of one of the models is investigated and priors for the distribution are suggested. PMID- 17117778 TI - Quantitative evaluation of three calibration methods for 3-D freehand ultrasound. AB - In this paper, three different calibration methods for three-dimensional (3-D) freehand ultrasound (US) are evaluated. Calibration is the process of estimating the rigid transformation from US image coordinates to the coordinate system of the tracking sensor mounted onto the probe. Calibration accuracy has an important impact on quantitative studies. Geometrical precision can also be crucial in many interventions and surgery. The proposed evaluation framework relies on a single point phantom and a 3-D US phantom which mimics the US characteristics of human liver. Four quality measures are used: 3-D point localization criterion, distance and volume measurements, and shape based criterion. Results show that during the acquisition procedure, volumetric measurements and shapes of the reconstructed object depend on probe motion used, particularly fan motions for which errors are larger. It is also shown that accurate calibration is essential to obtain reliable quantitative information. PMID- 17117779 TI - Towards a new tool for the evaluation of the quality of ultrasound compressed images. AB - This paper presents a new tool for the evaluation of ultrasound image compression. The goal is to measure the image quality as easily as with a statistical criterion, and with the same reliability as the one provided by the medical assessment. An initial experiment is proposed to medical experts and represents our reference value for the comparison of evaluation criteria. Twenty one statistical criteria are selected from the literature. A cumulative absolute similarity measure is defined as a distance between the criterion to evaluate and the reference value. A first fusion method based on a linear combination of criteria is proposed to improve the results obtained by each of them separately. The second proposed approach combines different statistical criteria and uses the medical assessment in a training phase with a support vector machine. Some experimental results are given and show the benefit of fusion. PMID- 17117780 TI - Using human and model performance to compare MRI reconstructions. AB - Magnetic resonance imaging (MRI) reconstruction techniques are often validated with signal-to-noise ratio (SNR), contrast-to-noise ratio, and mean-to-standard deviation ratio measured on example images. We present human and model observers as a novel approach to evaluating reconstructions for low-SNR magnetic resonance (MR) images. We measured human and channelized Hotelling observers in a two alternative forced-choice signal-known-exactly detection task on synthetic MR images. We compared three reconstructions: magnitude, wavelet-based denoising, and phase-corrected real. Human observers performed approximately equally using all three reconstructions. The model observer showed very close agreement with the humans over the range of images. These results contradict previous predictions in the literature based on SNR. Thus, we propose that human observer studies are important for validating MRI reconstructions. The model's performance indicates that it may provide an alternative to human studies. PMID- 17117781 TI - The importance of peer review. PMID- 17117782 TI - Applications of metabolomics in agriculture. AB - Biological systems are exceedingly complex. The unraveling of the genome in plants and humans revealed fewer than the anticipated number of genes. Therefore, other processes such as the regulation of gene expression, the action of gene products, and the metabolic networks resulting from catalytic proteins must make fundamental contributions to the remarkable diversity inherent in living systems. Metabolomics is a relatively new approach aimed at improved understanding of these metabolic networks and the subsequent biochemical composition of plants and other biological organisms. Analytical tools within metabolomics including mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy can profile the impact of time, stress, nutritional status, and environmental perturbation on hundreds of metabolites simultaneously resulting in massive, complex data sets. This information, in combination with transcriptomics and proteomics, has the potential to generate a more complete picture of the composition of food and feed products, to optimize crop trait development, and to enhance diet and health. Selected presentations from an American Chemical Society symposium held in March 2005 have been assembled to highlight the emerging application of metabolomics in agriculture. PMID- 17117783 TI - Simultaneous determination of ginsenosides and polyacetylenes in American ginseng root (Panax quinquefolium L.) by high-performance liquid chromatography. AB - A method for simultaneous determination of ginsenosides and polyacetylenes in Panax quinquefolium L. (American ginseng) roots was developed. The ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Ro, malonyl-Rb1, malonyl-Rc, and malonyl-Rd and the polyacetylenes falcarinol and panaxydol were extracted from fresh ginseng roots in a sequential extraction process with 100% methanol followed by 80% aqueous methanol and quantified simultaneously in extracts by high-performance liquid chromatography using diode array detection. Separations were achieved with a phosphate buffer-acetonitrile gradient system using an RP-C18 column. Except for Rd, the present extraction method resulted in similar or significantly higher concentrations of both ginsenosides and polyacetylenes in comparison to commonly used extraction methods for these compounds. The contents of polyacetylenes and ginsenosides were determined in the root hairs, lateral roots, and main roots of 6 year old ginseng plants. The total mean concentrations of ginsenosides and polyacetylenes in root hairs were 31.0 g/kg fresh weight (FW) and 2.6 g/kg FW, respectively, whereas the concentrations of these bioactive compounds in the main roots were significantly lower with total mean concentrations of 17.8 g/kg FW for ginsenosides and 0.6 g/kg FW for polyacetylenes. The concentration of individual and total ginsenosides and polyacetylenes did not differ significantly between main roots of different sizes. Consequently, it is possible to do quantitative screening for ginsenosides and polyacetylenes to breed ginseng roots with higher levels of bioactive compounds. PMID- 17117784 TI - Identification and characterization of conjugated linolenic acid isomers by Ag+ HPLC and NMR. AB - Interest in the biological activity of conjugated linolenic acids (CLnA) is growing. Technically, previous methods could not fully characterize and identify CLnA isomers. The present study is the first report on application of silver ion impregnated high-performance liquid chromatography (Ag+-HPLC) into separation of CLnA methyl ester (CLnAMe) mixture. Compared with the GC and reversed-phase HPLC techniques, Ag+-HPLC could resolve and separate CLnAMe isomers into three clusters of peaks namely ttt, ctt and ctc geometrical groups. Each positional isomer belonging to the same geometrical group could be further separated by Ag+ HPLC. Quantitative 13C NMR properties of CLnAMe mixture were subsequently studied. Olefinic signals in the CLnAMe mixture were assigned and used to quantify each isomer in combination with Ag+-HPLC. The present study provided a detailed analysis of a complex CLnA mixture and may have an important application in future studies dealing with synthesis, oxidative stability, and bioactivity of individual CLnA isomers. PMID- 17117785 TI - Enhancement effect study of some organic acids on the calcium availability of vegetables: application of the dynamic in vitro simulated gastrointestinal digestion method with continuous-flow dialysis. AB - The effect of added organic acids on the calcium availability of vegetables was investigated using the dialysis profiles obtained from an in vitro simulated gastrointestinal digestion with continuous-flow dialysis method. Citric acid was the most effective enhancer followed by tartaric, malic, and ascorbic acids. For amaranth, which has a low calcium availability (5.4%), a significant increase of availability was observed with increasing concentrations of all acids studied. With the continuous-flow dialysis approach, organic acids could be observed to promote the dialyzability even at an elevated intestinal pH. An enhancement effect from added organic acids was not clearly observed for Chinese kale, which itself contains a high amount of available calcium (52.9%). PMID- 17117786 TI - Quantitative analysis of tylosin in eggs by high performance liquid chromatography with electrospray ionization tandem mass spectrometry: residue depletion kinetics after administration via feed and drinking water in laying hens. AB - Maximum residue limits (MRLs) have been established by the European Union when tylosin is used therapeutically. They are fixed at 200 microg/kg for eggs. A highly sensitive and selective quantitative liquid chromatography electrospray ionization tandem mass spectrometry (LC/ESI/MS/MS) method suitable for monitoring tylosin residues in eggs to determine its depletion kinetics was developed and validated. For sample pretreatment all samples were liquid-liquid extracted with citrate buffer (pH 5.0) and acetonitrile. Liquid chromatographic separation was carried out on a reversed phase C18 column employing a 0.5% formic acid/acetonitrile gradient system. The tylosin recovery in eggs at a concentration range from 1.0-400 microg/kg was >82% with relative standard deviations between 1.5 and 11.0%. In two experimental studies administrating tylosin via feed (final dosage: 1.5 g/kg) or drinking water (final dosage: 0.5 g/L), no residues above the MRL were found during and after treatment. Moreover, all samples were well below the actual MRL of 200 microg/kg. Therefore, our residue data suggest that a withholding period for eggs is not required when laying hens are treated with tylosin in recommended dosages via feed or drinking water. KEYWORDS: Tylosin; residue; depletion; laying hen; withholding period; mass spectrometry. PMID- 17117787 TI - Analysis of the contents of pungent compounds in fresh Korean red peppers and in pepper-containing foods. AB - An HPLC method has been developed for the analysis of extracts of fresh peppers containing capsaicinoids and of both capsaicinoids and piperines in pepper containing foods produced and sold in Korea. The HPLC method was optimized by defining how composition of the mobile phase affected retention times. Both identification and quantification were based on retention times and the following criteria: linearity of the UV response at 280 nm in HPLC, recoveries from spiked samples, and observed individual molecular ions in the mass spectra of the extracts determined by liquid chromatography-mass spectrometry. This method, with a limit of detection of approximately 15-30 ng, was used to quantify the distribution of capsaicinoids in 11 Korean whole peppers and in 12 commercial pepper-containing foods. Total capsaicinoid levels of whole peppers ranged from 1.21 microg/g for the PR Gang ja variety to 121.1 microg/g for the Chung yang variety. The levels in food extracts, four of which also included two piperines, ranged from 11.0 microg/g for radish kimuchi to 3752 microg/g for capsaicin sauce. The results demonstrate (a) the usefulness of the HPLC method for the simultaneous analysis of capsaicinoids derived from red peppers and piperines derived from black and white peppers extracted from complex food matrices and (b) the wide-ranging spread of levels of pungent pepper compounds in fresh peppers and in pepper-containing foods consumed in Korea. PMID- 17117788 TI - Effects of solid-state enzymatic treatments on the antioxidant properties of wheat bran. AB - This study evaluated the potential of solid-state enzyme treatments to release insoluble bound antioxidants such as phenolic acids from wheat bran, thereby improving its extractable and potentially bioaccessible antioxidant properties including scavenging capacities against peroxyl (ORAC), ABTS cation, DPPH and hydroxyl radicals, total phenolic contents, and phenolic acid compositions. Investigated enzyme preparations included Viscozyme L, Pectinex 3XL, Ultraflo L, Flavourzyme 500L, Celluclast 1.5L, and porcine liver esterase. Results showed significant dose-dependent increases in extractable antioxidant properties for some enzyme preparations, and Ultraflo L was found to be the most efficient enzyme, able to convert as much as 50% of the insoluble bound ferulic acid in wheat bran to the soluble free form. The effect of moisture content on these solid-state enzyme reactions was also evaluated and found to be dependent on enzyme concentration. These data suggest that solid-state enzyme treatments of wheat bran may be a commercially viable post-harvest procedure for improving the bioaccessibility of wheat antioxidants. PMID- 17117789 TI - Antioxidant activity of piper betel leaf extract and its constituents. AB - The 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay of the ethanol extracts of three varieties (Bangla, sweet, and Mysore) of Piper betel (pan) revealed the Bangla variety to possess the best antioxidant activity that can be correlated with the total phenolic content and reducing powers of the respective extracts. Column chromatography of the extract of the Bangla variety led to the isolation of chevibetol (CHV), allylpyrocatechol (APC), and their respective glucosides. The HPTLC analyses of the extracts revealed similar chemical profiles in all three P. betel varieties, although the concentrations of CHV and APC were significantly less in the sweet and Mysore varieties. Among the isolated compounds, APC showed the best results in all the in vitro experiments. It could prevent Fe(II)-induced lipid peroxidation (LPO) of liposomes and rat brain homogenates as well as gamma ray-induced damage of pBR322 plasmid DNA more efficiently than CHV. The superior anti-LPO and radioprotective activities of APC vis-a-vis those of CHV could not be explained by their respective Fe(II) chelation and .OH radical scavenging capacities. The better ability of APC to scavenge O2-. radicals and H2O2 might account for the results. PMID- 17117790 TI - Effects of ingested turmeric oleoresin on glucose and lipid metabolisms in obese diabetic mice: a DNA microarray study. AB - Turmeric, the rhizome of Curcuma longa L., has a wide range of effects on human health. Turmeric oleoresin, an extract of turmeric, is often used for flavoring and coloring. Curcuminoids and turmeric essential oil are both contained in turmeric oleoresin, and both of these fractions have hypoglycemic effects. In the present study, we comprehensively assessed the effect of turmeric oleoresin on hepatic gene expression in obese diabetic KK-Ay mice using DNA microarray analysis and quantitative real-time polymerase chain reaction (PCR). Female KK-Ay mice aged 6 weeks (n = 6/group) were fed a high-fat diet containing turmeric oleoresin, curcuminoids, and essential oil for 5 weeks. The same diet without any of these fractions was used as a control diet. Ingestion of turmeric oleoresin and essential oil inhibited the development of increased blood glucose and abdominal fat mass, while curcuminoids only inhibited the increase in blood glucose. DNA microarray analysis indicated that turmeric oleoresin ingestion up regulated the expression of genes related to glycolysis, beta-oxidation, and cholesterol metabolism in the liver of KK-Ay mice, while expression of gluconeogenesis-related genes was down-regulated. Real-time PCR analysis was conducted to assess the contribution of the curcuminoids and essential oil in turmeric oleoresin to the changes in expression of representative genes selected by DNA microarray analysis. This analysis suggested that curcuminoids regulated turmeric oleoresin ingestion-induced expression of glycolysis-related genes and also that curcuminoids and turmeric essential oil acted synergistically to regulate the peroxisomal beta-oxidation-related gene expression induced by turmeric oleoresin ingestion. These changes in gene expression were considered to be the mechanism by which the turmeric oleoresin affected the control of both blood glucose levels and abdominal adipose tissue masses. All of these results suggest that the use of whole turmeric oleoresin is more effective than the use of either curcuminoids or the essential oil alone. PMID- 17117791 TI - Production of triacetylhydroxytyrosol from olive mill waste waters for use as stabilized bioantioxidant. AB - A hydroxytyrosol triacetyl derivative was very efficiently produced as a highly pure stabilized antioxidant compound by a short treatment of olive mill waste water (OMWW) organic extracts, rich in hydroxytyrosol, with an acetylating mixture composed of HClO4-SiO2 and Ac2O (Chakborti and Gulhane reaction), in mild and safe conditions. A successive single step of middle pressure liquid chromatography (MPLC) purification of the reaction product was performed, with an overall yield of 35.6%. (This process, including both the Chakborti and Gulhane reaction and the MPLC purification, is protected by an international patent under PCT/IT2005/000781.) The o-diphenol triacetyl derivative was also produced by direct reaction of hydroxytyrosol, previously purified by MPLC, with HClO4-SiO2 and Ac2O, with an overall yield of 29.5%. A further procedure for the production of the hydroxytyrosol triacetyl derivative was consistent with the direct treatment of raw OMWW with the acetylating agent and a single step of MPLC purification, with an overall yield of 27.6%. The purified natural triacetylhydroxytyrosol confirmed the same strong protective effects against the oxidative stress in human cells as the corresponding synthetic compound, likely because of the biochemical activation of the acetyl derivative into the active parent hydroxytyrosol by esterases. We therefore propose the utilization of OMWW for recovering hydroxytyrosol as a natural antioxidant in a chemically stabilized form, with a good yield, which can be potentially used as a nontoxic functional component in nutritional, pharmaceutical, and cosmetic preparations. PMID- 17117792 TI - Genetic engineering of Nicotiana tabacum for reduced nornicotine content. AB - Nornicotine is an undesirable secondary alkaloid in cultivated tobacco, because it serves as a precursor to N'-nitrosonornicotine (NNN), a tobacco-specific nitrosamine with suspected carcinogenic properties. Nornicotine is produced through the oxidative N-demethylation of nicotine by a nicotine N-demethylase enzyme during the senescence and curing of tobacco leaves. While the nornicotine content of most commercial burley tobacco is low, a process termed "conversion" can bestow considerably increased nornicotine levels in a portion of the plants within the population. Previously, we isolated a nicotine N-demethylase gene, designated CYP82E4, and demonstrated that RNAi-induced silencing of CYP82E4 and its close homologues is an effective means for suppressing nicotine to nornicotine conversion. In this study, we used real-time polymerase chain reaction to confirm the central role of CYP82E4 in nicotine N-demethylation by demonstrating that the transcript accumulation of CYP82E4 is enhanced as much as 80-fold in converter vs nonconverter tobacco. We also show the design of an optimized RNAi construct (82E4Ri298) that suppressed nicotine to nornicotine conversion from 98% to as low as 0.8% in a strong converter tobacco line, a rate of nornicotine production that is about 3.6-fold lower than typically detected in commercial varieties. Southern blot analysis showed that a single copy of the RNAi transgene was as effective in suppressing nornicotine accumulation as multiple copies. Greenhouse-grown transgenic plants transformed with the RNAi construct were morphologically indistinguishable from the empty vector or wild type controls. These results demonstrate that the genetic transformation of tobacco with the 82E4Ri298 construct is an effective strategy for reducing nornicotine and ultimately NNN levels in tobacco. KEYWORDS: Alkaloid; cytochrome P450; gene silencing; nicotine N-demethylase; N'-nitrosonornicotine; plant genetic engineering; metabolic engineering; Nicotiana tabacum L.; real-time PCR; RNA interference; tobacco-specific nitrosamines. PMID- 17117793 TI - Odor-active alcohols from the fungal transformation of alpha-farnesene. AB - Submerged microbial cultures were screened for their potential to oxifunctionalize alpha-farnesene. The major oxidation product in all transforming cultures, 3,7,11-trimethyldodeca-1,3(E),5(E)10-tetraen-7-ol, showed a pleasant citrus-like odor and peak concentrations of 170 mg L-1. An Aspergillus niger isolate from mango generated another two terpene alcohols identified as diastereomeric menth-1-en-3-[2-methyl-1,3-butadienyl]-8-ol, a new natural compound with an apricot-like odor. The regiospecifity of the oxygen attack with concurrent lack of stereoselectivity suggested that the initial step of the bioconversion resembled the chemical autoxidation starting with the generation of an intermediate resonance-stabilized carbon-centered radical or carbocation. PMID- 17117794 TI - Production of recombinant ScFv antibodies against methamidophos from a phage display library of a hyperimmunized mouse. AB - A recombinant phage display library was generated using splenocyte mRNA isolated from a Balb/c mouse hyperimmunized with a hapten conjugate that mimicked the structure of methamidophos, one of the most acutely toxic organophosphate pesticides. Three recombinant single-chain variable fragment (scFv) antibodies with the highest specificity for methamidophos, termed 28D4, 29D0, and 36B2, were produced via a stringent selection protocol. In a competitive enzyme-linked immunosorbent assay, the IC50 values for 28D4, 29D0, and 36B2 were 46.25, 35.39, and 17.99 ng/mL, respectively. The cross-reactivity of the three scFv antibodies with other organophosphate pesticides was below 0.1% except for acephate (O,S dimethyl acetylphosphoramidothioate). Nucleotide and deduced amino acid sequences indicated that the respective heavy chains and light chains of the three scFvs were involved in the distinctive VDJ segment rearrangements associated with somatic hypermutations during the process of several immunizations with higher dosages of immunogen. Taken together, these data constitute the first detailed description of an immunoassay that utilizes scFvs against the methamidophos, an analyte with a simple structure and low molecular mass (141 Da). PMID- 17117795 TI - Role of water upon the formation of acrylamide in a potato model system. AB - The moisture sorption isotherms of a commercial potato powder were investigated at 20 degrees C for water activities ranging from 0.11 to 0.97. The sorption isotherms were typical type-II sigmoidal curves, with a steep increase in moisture content for water activities above 0.9 and exhibiting hysteresis over the whole water activity range. On the basis of the isotherms, the influence of the initial water activity and moisture content on both Maillard browning and acrylamide formation was determined by heating oil containing potato powder mixtures in a closed stainless-steel tubular reactor. The Maillard browning, as determined spectrophotometrically, showed an optimum at intermediate water activities. The yields of acrylamide, expressed relatively to the molar amount of asparagine, remained constant below 0.8 aw and below moisture contents of about 20% (on a dry basis). For the more intense heat treatments, an increased acrylamide yield was however observed at higher moisture contents, with an optimum at water contents of about 100% (on a dry basis). However, this increase and optimum was not observed at less intense heat treatments. At moisture contents above 100%, a significant decrease in acrylamide yields was assessed, although the water activity increased only marginally in this area of the sorption isotherms. It was thus observed that the acrylamide content was rather dependent upon the moisture content than upon the water activity in the high moisture potato powder model system. PMID- 17117796 TI - Changes in the free amino acid contents of honeys during storage at ambient temperature. AB - This study was carried out to establish the changes in the free amino acid contents of floral honeys, honeydew honeys, and blend honeys during storage at room temperature and to test the capacity of the amino acids to distinguish the origin of the honeys after storage. For this purpose, 54 artisanal honeys (39 floral, 5 honeydew, and 10 blend) were studied. Samples were taken from recently collected honeys and at 3, 6, 9, 12, 16, 20, and 24 months after harvesting. The contents of most of the free amino acids were found to decrease with storage time, with the greatest reduction observed in the first 9 months. The contents of the amino acids aspartic acid, beta-alanine, and proline increased in the first few months after storage, reaching maximum values at 6 months, suggesting the possible existence of enzymatic activities. The application of stepwise discriminant analysis to the free amino acid content data demonstrated that the contents of the amino acids valine, beta-alanine, gamma-aminobutyric acid, serine, isoleucine, alpha-alanine, ornithine, and glutamine correctly assigned 87% of honeys to their group of origin: floral, honeydew, or blend. PMID- 17117797 TI - Glyceraldehyde bridging between flavanols and malvidin-3-glucoside in model solutions. AB - Hydroxyl radicals (.OH) seem to have an important role in the oxidation of wine constituents and the production of important electrophilic aldehydes and ketones. In this experiment, glyceraldehyde, a .OH oxidation product of glycerol, recently described in wine, reacts with (+)-catechin, (-)-epicatechin, and malvidin-3 glucoside (Mv3gl), in model solutions, yielding new condensed phenolic compounds. The adduct compounds formed were separated by means of reversed phase liquid chromatography and detected and characterized using UV-vis and electrospray ionization mass spectrometry. Flavanol-flavanol and anthocyanin-flavanol adducts linked with glyceraldehyde yielded compounds with m/z ratios for their main ions, in positive ion mode, of 653.2 for the (+)-catechin dimer or the (-)-epicatechin dimer and 855.5 for Mv3gl/(+)-catechin or Mv3gl/(-)-epicatechin dimers. The possible occurrence of these compounds in wine is suggested, and the potential role of these and related reactions in wine aging is discussed. PMID- 17117798 TI - Prolonged persistence of fecally excreted ivermectin from reindeer in a sub arctic environment. AB - In December 2001 and 2002, feces from reindeer calves treated with ivermectin were distributed on plots established on two types of forested reindeer pasture in northern Finland. The ungrazed plots were on an enclosure that had been fenced to prevent reindeer access for the last 6 years. The grazed plots were on an area that had been heavily stocked by reindeer during the last 5 years. After enclosures had been established, reindeer and large wildlife were prevented from entering by a fence. Topsoil samples (feces, vegetation, and soil) were collected monthly during the summers of the following 2 years, over a period of from 25 to 95 weeks after deposition. The samples were analyzed for ivermectin using HPLC. Although ivermectin degradation rapidly took place during the first spring, considerable residual ivermectin could be measured throughout the sampling time, showing that ivermectin in feces on pasture may not be photodegraded as rapidly as previously believed. The results support the need for further environmental evaluation studies on the use of ivermectin to control reindeer parasites. PMID- 17117799 TI - Methyl jasmonate induces defense responses in grapevine and triggers protection against Erysiphe necator. AB - Grapevine is subject to a number of diseases that affect yield and wine quality. To limit the excessive use of phytochemicals in the vineyard, alternative strategies have to be developed. Plant treatment with signaling molecules like elicitors stimulates their natural defense mechanisms. To improve grapevine tolerance against fungal pathogens, Vitis vinifera plants were treated with a natural exogenous elicitor, methyl jasmonate (MeJA). MeJA-treated leaves (Cabernet Sauvignon foliar cuttings) reacted by increasing transcript levels coding pathogenesis-related proteins (acidic class IV chitinase, serine protease inhibitor, polygalacturonase-inhibiting protein, and beta-1,3-glucanase) and coding enzymes involved in phytoalexin biosynthesis (one phenylalanine ammonia lyase and one stilbene synthase). This was correlated with the accumulation of stilbenes (antimicrobial compounds). The eliciting activity of MeJA was confirmed by enhanced tolerance of grapevine foliar cuttings and vineyard against powdery mildew (75% and 73%, respectively). On the basis of these original results, MeJA can therefore act as an efficient elicitor in an alternative strategy of grapevine protection. PMID- 17117800 TI - MCPA (4-Chloro-2-ethylphenoxyacetate) resistance in hemp-nettle (Galeopsis tetrahit L.). AB - The physiological basis for MCPA resistance in a hemp-nettle (Galeopsis tetrahit L.) biotype, obtained from a MCPA-resistant field population, was investigated. Dose-response studies revealed that the resistance factor for MCPA, based on GR50 comparisons of total dry weight of resistant (R) and susceptible (S) plants, was 3.3. Resistance factors for fluroxypyr, dicamba, 2,4-D, glyphosate, and chlorsulfuron were 8.2, 1.7, 1.6, 0.7, and 0.6, respectively. MCPA resistance was not due to differences in absorption, because both R and S biotypes absorbed 54% of applied [14C]MCPA 72 h after treatment. However, R plants exported less (45 vs 58% S) recovered 14C out of treated leaves to the apical meristem (6 vs 13% S) and root (32 vs 38% S). In both biotypes, approximately 20% of the 14C recovered in planta was detected as MCPA metabolites. However, less of the 14C recovered in the roots of R plants was MCPA. Therefore, two different mechanisms protect R hemp-nettle from MCPA phytotoxicity: a lower rate of MCPA translocation and a higher rate of MCPA metabolism in the roots. In support of these results, genetic studies indicated that the inheritance of MCPA resistance is governed by at least two nuclear genes with additive effects. PMID- 17117801 TI - Design and syntheses of novel phthalazin-1(2H)-one derivatives as acetohydroxyacid synthase inhibitors. AB - A series of 2-substituted-8-(4,6-dimethoxypyrimidin-2-yloxy)-4-methylphthalazin-1 one derivatives, 7a-7w, were designed via an ortho-substituent cyclization strategy to discover a new herbicidal lead structure. These compounds were synthesized by a seven-step route using 3-hydroxy-acetophenone as a starting material. Determination of the Ki values against wild-type A. thaliana acetohydroxyacid synthase (AHAS) (EC 4.1.3.18) indicated that some of the compounds displayed good enzyme inhibition activity comparable to that of KIH 6127. The further preliminary bioassay data on weeds showed that the synthesized compounds exhibited typical injury symptoms of AHAS-inhibiting herbicides, and some of them showed broad-spectrum and high herbicidal activities in postemergence treatments against Echinochloa crusgalli, Digitaria sanguinalis, Setaria viridis, Brassica juncea, Amaranthus retroflexus, and Chenopodium album at an application rate of 150 g ai/ha. To our knowledge, this is the first report of methylphthalazin-1-one derivatives as AHAS inhibitors. PMID- 17117802 TI - Diferulate content of maize sheaths is associated with resistance to the Mediterranean corn borer Sesamia nonagrioides (Lepidoptera: Noctuidae). AB - The leaf sheaths of selected inbred lines of maize (Zea mays L.) with variable levels of stem resistance to the Mediterranean corn borer Sesamia nonagrioides (Lefevbre) were evaluated for antibiotic effect on insect development. Phytochemical analyses of leaf sheaths were conducted for cell wall phenylpropanoid content to gain a better understanding of maize-resistance mechanisms. Laboratory bioassays established that sheath tissues from different genotypes significantly affected the growth of neonate larvae. Three hydroxycinnamates, p-coumaric, trans-ferulic, and cis-ferulic acids, and three isomers of diferulic acid, 8-5', 8-O-4', and 8-5' b (benzofuran form), were identified. Significant negative correlations were found between larvae weight and diferulic acid content for six genotypes. These results are in agreement with previous studies concerning the role of cell wall structural components in stem borer resistance. PMID- 17117803 TI - Enantiomeric differences in permethrin degradation pathways in soil and sediment. AB - Chirality occurs widely in synthetic pyrethroids. Studies have shown significant differences in both aquatic toxicity and degradation rates between enantiomers from the same diastereomer of selected pyrethroids. To better understand chiral selectivity in biodegradation of pyrethroids, 14C-labeled permethrin was used to characterize enantiomeric differences in the formation of transformation intermediates in two soils and a sediment. Individual enantiomers of permethrin were spiked into soil and sediment samples, and transformation products were identified with known standards. Enantioselectivity was observed in most treatments when the dissipation of the parent enantiomers, the amount of intermediates and bound residues formed, and mineralization rates were compared between the enantiomers. The results show that all enantiomers of permethrin hydrolyzed rapidly and that the hydrolysis products were quickly further transformed. The direct hydrolysis products, cyclopropanic acid (Cl2CA), 3 phenoxybenzyl alcohol (PBalc), and 3-phenoxybenzoic acid (PBacid), were recovered at small percentages, ranging from 1 to 14% for Cl2CA and from 0.2 to 6% for PBalc and PBacid. The R-enantiomer of both cis- and trans-permethrin was mineralized more quickly than the S-enantiomer after hydrolysis. The degradation products from cis-permethrin were more persistent than those from trans permethrin. As some transformation intermediates of permethrin may have greater acute and chronic toxicity than the parent compound, enantioselectivity in the formation of degradation intermediates may lead to different overall toxicities and merit further investigation. This study suggests that for chiral compounds, enantioselectivity may be reflected not only in the dissipation of the parent enantiomers but also in the kinetics of formation of intermediate transformation products. PMID- 17117804 TI - Lysimeter experiment to investigate the potential influence of diffusion-limited sorption on pesticide availability for leaching. AB - Pesticide leaching from soil has been shown to decrease with increasing time from application to irrigation. It is hypothesized that the availability of compounds for leaching decreases due to diffusion and sorption inside soil aggregates. Previous work showed that pesticide sorption inside soil aggregates increases significantly during the first days after application. The study presented here tested if diffusion into aggregates could explain the leaching of four aged pesticides from manually irrigated soil cores. Azoxystrobin, chlorotoluron, cyanazine, and bentazone were applied to 30 undisturbed cores (25 cm long, 23.7 cm diameter) from a clay loam soil. The soil cores were irrigated 1, 3, 7, 14, and 28 days after application. Leachate was collected and analyzed. The amount of pesticide found in leachate decreased rapidly with time from application. Pesticide losses in leachate declined 2.5-27 times faster than total residues in soil. The decline was 4-5 times faster for the more strongly sorbed pesticides (azoxystrobin, chlorotoluron, and cyanazine) than for bentazone. In previous work, we derived a model to describe sorption and diffusion of the pesticides in small aggregates from the same soil. The diffusion model was used here to describe sorption inside the large aggregates in the soil cores and extended to describe pesticide leaching by interaggregate flow. The model showed a significant decline in leaching with time from application, which supports the theory that diffusion-limited sorption in aggregates influences the availability for pesticide leaching, although it does not exclude alternative explanations for this decline. The model well described the decline in leaching for three out of four pesticides. The interaggregate transport model could, however, not account for the amount of preferential flow in the cores and underestimated the leaching of bentazone. PMID- 17117805 TI - Photosensitizing properties of protein hydrolysate-based fertilizers. AB - The use of protein hydrolysate-based fertilizers (PHF) as adjuvant for pesticides or herbicides has been proposed; however, the behaviors of mixtures of PHFs and pesticides under solar light are not known, and various photochemical reactions may occur. The photosensitizing properties of PHFs were investigated in water solutions (0.8 g of total organic carbon L(-1)) within the wavelength range of 300-450 nm, using furfuryl alcohol (FA) as a probe to test the involvement of singlet oxygen and Irgarol 1051 as an example of organic pollutant. Two commercial PHFs and one standard PHF were studied, all of the products being of animal origin. PHFs photosensitize the transformation of FA (10(-4) M), and the kinetics of FA disappearance follows an apparent first-order rate law. Through the use of sodium azide (1 x 10(-3) M) as singlet oxygen scavenger and deuterium oxide (D2O) for increasing the singlet oxygen lifetime it was shown that singlet oxygen contributes largely to the phototransformation of FA. The replacement of water by D2O increases the apparent first-order rate constant 6 times, whereas the addition of sodium azide reduces it by approximately 90%. These results are confirmed using Irgarol 1051 (10(-5) M). The photosensitizing properties of PHFs might be due to pigments naturally present in tissues from which they are extracted or to compounds generated during the production processes. PMID- 17117806 TI - Effect of copper on the volatility of aroma compounds in a model mouth system. AB - Copper is thought to influence aroma perception by affecting volatility of aroma compounds in the mouth through interaction with salivary components, especially proteins. Our objective was to identify the effect of copper on the volatility of aroma compounds and the role of copper-protein interaction in volatile chemistry in the mouth. Copper (2.5 mg/L) and four aroma compounds (hexanal, butyl acetate, 2-heptanone, and ethyl hexanoate, 0.5 microL/L each) were added to model systems containing water, electrolytes, and artificial saliva at different pH levels. Headspace concentration of each volatile was measured using SPME-GC analysis. Copper in the model systems increased headspace concentration of volatiles at pH 6.5, but no change in volatility was observed at pH 7.0. At pH 7.5, the presence of copper in the artificial saliva system containing mucin and alpha-amylase decreased headspace volatile concentration, whereas histatin did not cause any changes in volatility. Effect of copper on volatiles at pH 6.5 may be due to increased solubility of copper at lower pH. Salivary proteins seem to interact with copper at pH 7.5. The interaction may change configuration of binding sites for aroma compounds in mucin. PMID- 17117807 TI - Determination of haloanisols in white wine by immunosorbent solid-phase extraction followed by enzyme-linked immunosorbent assay. AB - A high through-put screening immunochemical method to control the presence of 2,4,6-trichloroanisol (TCA) and 2,4,6-tribromoanisol (TBA), the main agents responsible for the musty odor in wine samples, has been developed. The method involves a selective (antibody-antigen) solid-phase extraction (SPE), followed by enzyme-linked immunosorbent assay (ELISA) analysis. The sample preparation method established uses for immunosorbents (ISs) prepared by covalently coupling antibodies developed for TCA on a sepharose support. At present, about 200-400 ng L-1 of TBA and TCA can be detected in white wine samples by the IS-SPE-ELISA method described here without any preconcentration step. Simultaneous analyses of many samples are possible with this method. Related chloroanisoles (2,3- and 2,6 dichloroanisols and 2,3,4,5-tetrachloroanisol) and chlorophenols (2,3,4,6 tetrachlorophenol and pentachlorophenol) usually present in contaminated wine samples are also effectively retained by the IS, although only 2,4,6-TCA and 2,4,6-TBA are detected by the ELISA used. The immunopurification procedure developed could also be useful as a selective cleanup method prior to chromatographic analysis. PMID- 17117808 TI - Effect of high-pressure-moderate-temperature processing on the volatile profile of milk. AB - The effects of high hydrostatic pressure on volatile generation in milk were investigated in this study. Raw milk samples were treated under different pressures (482, 586, and 620 MPa), temperatures (25 and 60 degrees C), and holding times (1, 3, and 5 min). Samples submitted to heat treatments alone (25, 60, and 80 degrees C for 1, 3, and 5 min) were used for comparison. Trace volatile sulfur compounds were analyzed using solid-phase microextraction (SPME) and gas chromatography (GC) with pulsed-flame photometric detection (PFPD), whereas the rest of the volatile compounds were analyzed using SPME-GC with flame ionization detection (FID). Multivariate analysis of variance (MANOVA) and principal component analysis (PCA) were used to study the effect of pressure, temperature, and time on volatile generation. Relative concentration increases of 27 selected volatile compounds were compared to an untreated sample. It was found that pressure, temperature, and time, as well as their interactions, all had significant effects (P < 0.001) on volatile generation in milk. Pressure and time effects were significant at 60 degrees C, whereas their effects were almost negligible at 25 degrees C. The PCA plot indicated that the volatile generation of pressure-heated samples at 60 degrees C was different from that of heated alone samples. Heat treatment tended to promote the formation of methanethiol, hydrogen sulfide, methyl ketones, and aldehydes, whereas high-pressure treatment favored the formation of hydrogen sulfide and aldehydes. PMID- 17117809 TI - Characterization of some mushroom and earthy off-odors microbially induced by the development of rot on grapes. AB - Grape rot is one of the major causes of degradation of many grape components and, thus, of deterioration in wine quality. In particular, the association of Botrytis cinerea with other, less visible, fungi frequently leads to the development of organoleptic defects in grapes and sometimes in wines. This study examines the nature of the volatile compounds responsible for mushroom, mossy, or earthy odors detected by gas chromatography-olfactometry in organic extracts of rotten grapes and musts. 2-Methylisoborneol, (-)-geosmin, 1-octen-3-one, 1-octen 3-ol, 2-octen-1-ol, and 2-heptanol were identified or tentatively identified. Their concentrations in musts were determined, and the impact of alcoholic fermentation by the yeast Saccharomyces cerevisiae was studied. The ability of fungi isolated from rotten grapes (Botrytis cinerea; Penicillium species including P. brevicompactum, P. expansum, P. miczynskii, P. pinophilum, P. purpurogenum, and P. thomii; Aspergillus section nigri; Rhizopus nigricans; and Coniothyrium sp.) to produce some of the identified compounds was evidenced. PMID- 17117810 TI - New constituents related to 3-methyl-2,4-nonanedione identified in green tea. AB - The volatile constituents of two exquisite green tea varieties, Kiyosawa tea from Japan and Long Jing tea from China, were investigated in order to identify new compounds responsible for the characteristic flavor of a green tea brew. The extracts were prepared by solid-phase extraction using Oasis-HLB-cartridges. Besides the common compounds of green tea chemistry, the already described compounds 3-methyl-2,4-nonanedione (1) and 3-hydroxy-3-methyl-2,4-nonanedione (2), products of degradation of furan fatty acids, as well as three new compounds related to compound 1 were identified. These were 1-methyl-2-oxopropyl hexanoate (3), 1-methyl-2-oxoheptyl acetate (4) and 2-butyl-4,5-dimethyl-3(2H)-furanone (5). Their syntheses and spectroscopic data are reported. Compound 2 increases the sweet, creamy aroma and the characteristic mouthfeel of a green tea flavor, compounds 3 and 4 contribute to its floral, juicy notes and compound 5 exhibits an interesting sweet, buttery flavor. PMID- 17117811 TI - Anatomical, histological, and histochemical changes in grape seeds from Vitis vinifera L. cv Cabernet franc during fruit development. AB - Cabernet franc berries were sampled at five stages from berry set to harvest from an experimental vineyard in mid Loire Valley. Seeds were collected from representative berries in term of stage of development. The evolution of seed was followed both macro- and microscopically. For microscopy analysis, seeds were cut, put in a fixation solution, and cut into thin sections with a microtome. Five staining solutions were used for each seed sample: toluidine blue O, phloroglucinol, periodic acid-Schiff's reagent and naphtol blue black, vanillin, and p-dimethylaminocinnamaldehyde. Toluidine blue O staining revealed the evolution of tissue structures during grape seed development. We studied the changes in chemical compounds (lignin, polysaccharides, proteins, and tannins) with the other reagents. Seed lignification was achieved at veraison. Proanthocyanidins were localized in epidermis, inner cells of the soft seed coat, and inner cell layer of the inner integument. Finally, the localization of flavan 3-ols was linked with changes in cell walls of the outer integument. PMID- 17117812 TI - Detection of free radical transfer in lipoxygenase I-B-catalyzed linoleic acid soybean protein interaction by electron spin resonance spectroscopy (ESR). AB - Effects of lipoxygenase I-B (LOX)-catalyzed oxidation of linoleic acid on soybean proteins was evaluated by electron spin resonance (ESR) and fluorescence spectroscopy in different model systems in the presence or absence of antioxidants. A strong central singlet signal was detected by ESR spectroscopy and identified as the carbon radical (g value range 2.0041-2.0054). A downfield shoulder attributed to the sulfur radical (g value 2.019-2.028) was also observed. The changes in soybean proteins were accompanied by an increase in fluorescence, indicating the formation of cross-links. Natural antioxidants such as ascorbic acid and alpha-tocopherol as well as synthetic antioxidants butyl hydroxytoluene (BHT) inhibited the development of both the free radical signal and the fluorescence when added to soybean proteins prior to incubation with linoleic acid and lipoxygenase I-B; the central singlet signal attributed to the carbon radical was reduced by 35-65%. This paper clearly indicates direct free radical transfer from oxidizing linoleic acid catalyzed by LOX to soybean proteins. PMID- 17117813 TI - Non-concordance between genetic profiles of olive oil and fruit: a cautionary note to the use of DNA markers for provenance testing. AB - To investigate the contribution of paternal alleles to the DNA content of olive oil, genetic analyses of olive DNA samples from fruits, leaves, and oil derived from the same tree (cv. Leccino) were carried out. DNA extracted from maternal tissues--leaves and flesh--from different fruits showed identical genetic profiles using a set of DNA markers. Additional simple sequence repeat (SSR) alleles, not found in the maternal samples, were amplified in the embryos (stone), and they were also detected in DNA extracted from the paste obtained by crushing whole fruits and from the oil pressed from this material. These results demonstrate that the DNA profile obtained from olive oil is likely to represent a composite profile of the maternal alleles juxtaposed with alleles contributed by various pollen donors. Therefore, care needs to be taken in the interpretation of DNA profiles obtained from DNA extracted from oil for resolving provenance and authenticity issues. PMID- 17117814 TI - Gene isolation, analysis of expression, and in vitro synthesis of glutathione S transferase from orange fruit [Citrus sinensis L. (Osbeck)]. AB - Glutathione S-transferases (GSTs) (EC 2.5.1.18) are ubiquitous enzymes that have a defined role in xenobiotic detoxification, but a deeper knowledge of their function in endogenous metabolism is still lacking. In this work, we isolated the cDNAs as well as the genomic clones of orange GSTs. Having considered gene organization and homology data, we suggest that the isolated GST gene is probably involved in the vacuolar import of anthocyanins. We also found that the blood and blond orange GSTs shared the same nucleotide sequences, but as expected, the GST expression in the nonpigmented orange cultivar [Citrus sinensis L. (Osbeck)] (Navel and Ovale) was strongly reduced as compared to that of the pigmented orange (Tarocco). Interestingly, in the crude extracts of pigmented orange fruits, the GST activity was reproducibly detected by providing either 1-chloro 2,4 dinitrobenzene (CDNB) or cyanidin-3-O-glucoside (C-3-G) as substrates; moreover, we have shown that cyanidin-3-O-glucoside acted as a powerful competitive inhibitor of 1-chloro-2,4 dinitrobenzene conjugation to reduced glutathione (GSH) in the pigmented orange, confirming that this molecule might easily bind to the active site of the enzyme and functions as a putative substrate. In addition, we have reported here the successful in vitro expression of orange GST cDNAs leading to a GST enzyme that is active against cyanidin-3-O glucoside, thus suggesting the probable involvement of the isolated gene in the tagging of anthocyanins for vacuolar import. This last result will help to study the kinetic and structural properties of orange fruit GST avoiding time-consuming protein purification procedures. PMID- 17117815 TI - Soluble starch synthase I effects differences in amylopectin structure between indica and japonica rice varieties. AB - The effect of soluble starch synthase I (SSI) on differences of amylopectin structure between the indica and japonica rice varieties was investigated. Native PAGE/active staining analysis showed that the SSI activity of an indica rice variety, "Kasalath", was significantly lower than that of a japonica rice variety, "Nipponbare", and that the low activity in "Kasalath" was maintained during seed development. The result of northern blot analyses suggests that the low expression of SSI in "Kasalath" is controlled at the transcription levels of SSI mRNA. Chain length distribution of amylopectin in F3 endosperms derived from a cross between two varieties showed that not only SSIIa but also SSI regulated the population of short chains. These results indicate that the low activity of SSI gives rise to the decrease of short chains in amylopectin of indica rice varieties, suggesting that SSI effects the differences in physicochemical properties between two varieties. PMID- 17117816 TI - Effect of dietary conjugated linoleic acid and monounsaturated fatty acid content on pig muscle and adipose tissue lipase and esterase activity. AB - Three levels (0%, 1%, and 2%) of conjugated linoleic acid (CLA) were combined with two levels (low and high) of monounsaturated fatty acids (MUFA) for pig feeding. The activity of neutral lipase (NL), acid lipase (AL), phospholipase (PL), neutral esterase (NE), and acid esterase (AE) was measured in extracts from muscle and subcutaneous adipose tissues. The addition of CLA in the diet only affected the lipolytic activity in muscle, whereas differences in MUFA content of pig diets were mainly responsible for the lipolytic enzyme modifications observed in adipose tissue. Nevertheless, a significant effect of the interaction CLA x MUFA on the activity of several lipolytic enzymes was observed in both tissues. The effect of either linoleic acid (LA) or CLA on the activity of muscle and adipose lipolytic enzymes was determined by in vitro assays. Remarkable inhibitory or activation effects were detected depending on the enzyme and kind of tissue. PMID- 17117817 TI - Lipid-oxidation-induced carboxymyoglobin oxidation. AB - We evaluated the usefulness of the ratio A503/A581 as a browning index (BI) for estimating brown color formation in solutions containing oxymyoglobin (OxyMb) and carboxymyoglobin (COMb). In split-chamber cuvette analyses with different proportions of metmyoglobin (MetMb), COMb and OxyMb, BI was highly correlated (r = 0.93-0.94) with direct estimation of MetMb. Moreover, A503/A581 was not influenced by different COMb-OxyMb proportions. Second, we investigated 4-hydroxy 2-nonenal (HNE)-induced spectral changes in OxyMb and COMb solutions. At pH 7.4 and 37 degrees C, BI was greater in HNE-treated OxyMb and COMb samples than in aldehyde-free controls (P < 0.05). However, at pH 5.6 and 4 degrees C, HNE induced browning was more pronounced in COMb than in OxyMb. These results indicated that COMb is susceptible to lipid-oxidation-induced browning in a pH- and temperature-dependent manner. PMID- 17117818 TI - Kaempferol in red and pinto bean seed (Phaseolus vulgaris L.) coats inhibits iron bioavailability using an in vitro digestion/human Caco-2 cell model. AB - Four different colored beans (white, red, pinto, and black beans) were investigated for factors affecting iron bioavailability using an in vitro digestion/human Caco-2 cell model. Iron bioavailability from whole beans, dehulled beans, and their hulls was determined. The results show that white beans contained higher levels of bioavailable iron compared to red, pinto, and black beans. These differences in bioavailable iron were not due to bean-iron and bean phytate concentrations. Flavonoids in the colored bean hulls were found to be contributing to the low bioavailability of iron in the non-white colored beans. White bean hulls contained no detectable flavonoids but did contain an unknown factor that may promote iron bioavailability. The flavonoids, kaempferol and astragalin (kaempferol-3-O-glucoside), were identified in red and pinto bean hulls via HPLC and MS. Some unidentified anthocyanins were also detected in the black bean hulls but not in the other colored bean hulls. Kaempferol, but not astragalin, was shown to inhibit iron bioavailability. Treating in vitro bean digests with 40, 100, 200, 300, 400, 500, and 1000 microM kaempferol significantly inhibited iron bioavailability (e.g., 15.5% at 40 microM and 62.8% at 1000 microM) in a concentration-dependent fashion. Thus, seed coat kaempferol was identified as a potent inhibitory factor affecting iron bioavailability in the red and pinto beans studied. Results comparing the inhibitory effects of kaempferol, quercitrin, and astragalin on iron bioavailability suggest that the 3',4'-dihydroxy group on the B-ring in flavonoids contributes to the lower iron bioavailability. PMID- 17117819 TI - Mechanical, barrier, and antimicrobial properties of apple puree edible films containing plant essential oils. AB - Edible films, as carriers of antimicrobial compounds, constitute an approach for incorporating plant essential oils (EOs) onto fresh-cut fruit surfaces. The effect against Escherichia coli O157:H7 of oregano, cinnamon, and lemongrass oils in apple puree film-forming solution (APFFS) and in an edible film made from the apple puree solution (APEF) was investigated along with the mechanical and physical properties of the films. Bactericidal activities of APFFS, expressed as BA50 values (BA50 values are defined as the percentage of antimicrobial that killed 50% of the bacteria under the test conditions) ranged from 0.019% for oregano oil to 0.094% for cinnamon oil. Oregano oil in the apple puree and in the film was highly effective against E. coli O157:H7. The data show that (a) the order of antimicrobial activities was oregano oil > lemongrass oil > cinnamon oil and (b) addition of the essential oils into film-forming solution decreased water vapor permeability and increased oxygen permeability, but did not significantly alter the tensile properties of the films. These results show that plant-derived essential oils can be used to prepare apple-based antimicrobial edible films for various food applications. PMID- 17117820 TI - Mycotoxins and other secondary metabolites produced in vitro by Penicillium paneum Frisvad and Penicillium roqueforti Thom isolated from baled grass silage in Ireland. AB - Secondary metabolites produced by Penicillium paneum and Penicillium roqueforti from baled grass silage were analyzed. A total of 157 isolates were investigated, comprising 78 P. paneum and 79 P. roqueforti isolates randomly selected from more than 900 colonies cultured from bales. The findings mostly agreed with the literature, although some metabolites were not consistently produced by either fungus. Roquefortine C, marcfortine A, and andrastin A were consistently produced, whereas PR toxin and patulin were not. Five silage samples were screened for fungal metabolites, with two visually moldy samples containing up to 20 mg/kg of roquefortine C, mycophenolic acid, and andrastin A along with minor quantities (0.1-5 mg/kg) of roquefortines A, B, and D, festuclavine, marcfortine A, and agroclavine. Three visually nonmoldy samples contained low amounts of mycophenolic acid and andrastin A. The ability of both molds to produce a diverse range of secondary metabolites in vitro and in silage should be a concern to livestock producers. PMID- 17117824 TI - Preparation of hierarchically porous nickel from macroporous silicon. AB - A novel approach for preparing hierarchical porous nickel structures has been developed. It is an aqueous processing of the template, macroporous silicon, which was etched in HF solution to have straight channels. By immersion in a prepared nickel solution, the silicon sidewalls were replaced by nickel deposits to form a total metallic structure. Since silicon framework was gradually consumed during the displacement reactions, the deposited nickel displayed additional feature of being highly porous on the sidewalls. With the initial straight microchannels, the metal structure was hierarchically porous. PMID- 17117825 TI - Coiled molecules in spring loaded devices. AB - A cylindrical capsule containing a coiled alkane incorporates spacers. The alkane lengthens and gauche strain is relieved. The spacer can be removed to reset the system to the original state. The system represents a spring-loaded device that operates reversibly in response to acids and bases. PMID- 17117826 TI - Compaction dynamics of single DNA molecules under tension. AB - Using transverse magnetic tweezers, we studied the dynamics of DNA compaction induced by hexaammine cobalt chloride under constant forces. Discontinuous DNA compaction events were found for forces ranging from 0.5 to 1.7 pN, with approximately 270 nm DNA adsorbed in each compaction event. Forces larger than 6 pN were found able to unravel the toroid in a similar intermittent stepwise manner. The observations indicate that the folding/unfolding events are transitions between two metastable structural states which are separated by a tension-dependent energy barrier. Analysis of the waiting time revealed that the degree of the package ordering of DNA in a toroid depends on the compaction kinetics. PMID- 17117827 TI - 13C direct detected NMR increases the detectability of residual dipolar couplings. AB - 13C direct detection is becoming an increasingly efficient approach to identify signals of residues that escape detection in 1H detected experiments. Pulse sequences have been developed to obtain 1H partially recoupled experiments for the measurement of the 1JHalphaCalpha and 1JHN couplings with the same resolution available in conventional 1H detected experiments. A consistent set of backbone rdc obtained without any 1H-based experiment has been obtained and shown to be effective for protein solution structure determination. PMID- 17117828 TI - Symmetrical gas-phase dissociation of noncovalent protein complexes via surface collisions. AB - Previous gas-phase dissociation experiments of protein-protein complexes have resulted in product ion distributions that are asymmetric by charge and mass, providing limited insight into the chemical nature of subunit organization and interaction. In these experiments, a symmetric charge distribution results from an "energy sudden" collision of protein-protein complexes with a surface, indicating that it may be possible to probe the suboligomeric structure of noncovalent complexes in the gas phase. It is proposed that energy sudden surface activation of cytochrome C homodimers results in dissociation without significant unfolding of one of the monomeric subunits. Previously proposed mechanisms for the dissociation of protein-protein complexes are discussed in the context of these results. These experiments demonstrate the potential to preserve the structural details of subunit interaction within a protein-protein complex and help elucidate the asymmetric nature of macromolecular dissociation in the gas phase. PMID- 17117829 TI - Ionic-like behavior of oppositely charged nanoparticles. AB - Mixtures of oppositely charged nanoparticles of various sizes and charge ratios precipitate only at the point of electroneutrality. This phenomenon-specific to the nanoscale and reminiscent of threshold precipitation of ions-is a consequence of the formation of core-and-shell nanoparticle aggregates, in which the shells are composed of like-charged particles and are stabilized by efficient electrostatic screening. PMID- 17117830 TI - Oxidative cross-coupling through double transmetallation: surprisingly high selectivity for palladium-catalyzed cross-coupling of alkylzinc and alkynylstannanes. AB - Two different organometallic reagents now can cross-couple together with the oxidative cross-coupling strategy. Palladium catalyzed oxidative cross-couplings of alkylzinc and alkynylstannane reagents using desyl chloride as the oxidant have been explored, which produce the desired Csp-Csp3 cross-coupling product in surprisingly high selectivity and yields. The current catalytic system tolerates the presence of beta-H, and the reactions using long chain alkyl zinc reagents gave the cross-coupling product in excellent yields and selectivities. PMID- 17117831 TI - Iron-catalyzed carbometalation of propargylic and homopropargylic alcohols. AB - Nucleophilic addition to alkynes represents an attractive approach to the synthesis of olefins. Obstacles to this strategy include the low reactivity of alkynes toward many organometallic reagents and difficulties associated with controlling the regioselectivity of addition. Here we demonstrate that Fe(III) salts are effective precatalysts for the carbometalation of alkynes. Primary and secondary propargylic and homopropargylic alcohols react with alkyl and aryl Grignard reagents to provide Z-allylic and -homoallylic alcohols as single stereo and regioisomers. Alkylation and arylation occur distal to the alcohol. Common oxygen protecting groups and tertiary nitrogens are tolerated. The intermediate vinyl magnesium or iron species can be trapped with a variety of electrophiles including aldehydes, allyl bromide, and N-bromosuccinimide. Diyne substrates undergo an unusual addition/cyclization reaction to generate cyclic dienes. A brief discussion of mechanism is included. PMID- 17117832 TI - Thickness-dependent structural transitions in fluorinated copper-phthalocyanine (F16CuPc) films. AB - The detailed structure of F16CuPc films on SiO2 has been determined by means of in situ grazing incidence X-ray diffraction from the first monolayer to thicker films. In contrast to films of the homologous H16CuPc molecule, the F16CuPc films exhibit the same structure independently from the deposition temperature. The films show a thickness-dependent polymorphism manifested in the in-plane crystal structure, which implies large differences in the molecular tilt within the cofacial stacking of the molecules. PMID- 17117833 TI - One-step microwave preparation of well-defined and functionalized polymeric nanoparticles. AB - Well-defined colloidal polymeric nanoparticles are important in advanced biomedical and optical technologies. We report a facile microwave methodology to prepare narrowly dispersed cross-linked polymeric nanoparticles at high solids content through a surfactant-free emulsion polymerization process. The nanoparticle size was controlled by using cross-linkers with enhanced reactivity through a one-step microwaving process, significantly simplifying the nanoparticle synthetic process. The successful size control was realized by confining the cross-linking to intraparticle cross-linking rather than interparticle cross-linking. We also discovered that the superheating/dielectric heating effect associated with microwave irradiation could be utilized to effectively reduce the nanoparticle size. PMID- 17117834 TI - Transient terminal Cu-nitrene intermediates from discrete dicopper nitrenes. AB - Reaction of the copper(I) beta-diketiminate {[Me3NN]Cu}2(mu-toluene) with the aryl azide N3Ar (Ar = 3,5-Me2C6H3) in toluene results in immediate effervescence and formation of the dicopper nitrene {[Me3NN]Cu}2(mu-NAr) (2) in 77% yield. The X-ray structure of 2 shows nearly symmetric bonding of the nitrene to two Cu centers separated by 2.911(1) A with Cu-N distances of 1.794(5) and 1.808(5) A along with a Cu-N-Cu angle of 107.8(2) degrees . This structure is conceptually related to the dicopper carbenes {[MexNN]Cu}2(mu-CPh2) (x = 2 or 3) (Dai, X.; Warren J. Am. Chem. Soc. 2004, 126, 10085. Badiei, Y. M.; Warren J. Organomet. Chem. 2005, 690, 5989.) which exhibit shorter Cu-Cu distances (2.4635(7) or 2.485(1) A) and acute Cu-C-Cu angles (79.51(14) or 80.1(2) degrees ). Addition of the Cu(I) anilidoimine {[Me2AI]Cu}2 (prepared from CuOtBu and the aniline-imine H[Me2AI] in 77% yield) to a benzene-d6 solution of 2 results in the formation of two new anilidoimine complexes {[Me2AI]Cu(mu- NAr)Cu[Me3NN] (5) and {[Me2AI]Cu}2(mu-NAr) (6) as well as [Me3NN]Cu(benzene) over 3 h. These observations are consistent with the slow dissociation of a [Me3NN]Cu fragment from 2 to generate the transient terminal nitrenes [Me3NN]Cu=NAr and [Me2AI]Cu=NAr quickly trapped by the [Me2AI]Cu fragment to form the new unsymmetrical and symmetrical dicopper nitrenes 5 and 6. Preliminary reactivity studies indicate electrophilic reactivity at the nitrene moiety. Dicopper nitrene 2 reacts with 10 equiv PMe3 and CNtBu to give ArN=PMe3 and ArN=C=NtBu in 94% and 92% yields, respectively, with concomitant formation of [Me3NN]Cu(L) (L = PMe3 and CNtBu). Reaction between 2 and 2 equiv PMe3 allows for observation of the structurally characterized Cu(I) phosphaimide [Me3NN]Cu(ArN=PMe3) (7). PMID- 17117835 TI - Increased alignment of electronic polymers in liquid crystals via hydrogen bonding extension. AB - Crucial for the development of enhanced electrooptic materials is the construction of highly anisotropic materials. Nematic liquid crystals are able to control the chain conformation and alignment of poly(phenylene ethynylene)s (PPEs), producing electronic polymers with chain-extended planar conformations for improved transport properties. Here, we show that the dichroic ratio, and hence polymer alignment, increases dramatically when interpolymer interactions are introduced by end capping the PPE with hydrogen bonding groups. This increased order can be readily turned off by the introduction of a competing monofunctionalized hydrogen bonding compound. The formation of hydrogen bonds between the polymers results in the formation of gels and elastomers which may be of interest for future applications. PMID- 17117836 TI - Frustrated organic solids display unexpected gas sorption. AB - Between nonsorptive solvate and desolvate forms of p-tert-butylcalix[5]arene lies a frustrated crystalline region in which there is rapid sorption of gases. From sorption studies, we conclude that the frustrated form is porous and that the intermediate form represents a new type of material that was previously unrecognized by traditional gas sorption trends or ideals for other active systems. PMID- 17117837 TI - Self-organized interconnect method for molecular devices. AB - Lack of an appropriate method for wiring molecules that have controlled functions and structures has been a barrier for the development of molecular devices. We developed an interconnect method to program three kinds of component molecules with their own functions and to wire a molecular device in a self-organized manner. By using the interconnect method we developed, we produced conductive wires and optical switching devices and have demonstrated their device functions. Our interconnect method allows us to control various molecular device characteristics by combining the three molecules. PMID- 17117838 TI - A controlled-release strategy for the generation of cross-linked hydrogel microstructures. AB - Microscale hydrogels of controlled sizes and shapes are useful for cell-based screening, in vitro diagnostics, tissue engineering, and drug delivery. However, the rapid cross-linking of many chemically and pH cross-linkable hydrogel materials prevents the application of existing micromolding techniques. In this work we present a method for fabricating micromolded calcium alginate and chitosan structures through controlled release of the gelling agent from a hydrogel mold. Replica molding was employed to generate patterned membranes, whereas microtransfer molding was used to produce microparticles of controlled shapes. To explore the viability of this technique for producing complex tissue engineering micro-architectures, this approach was used to generate cell-laden size- and shape-controlled 3D microgels as well as composite hydrogels with well defined spatially segregated regions. In addition, shape-controlled microstructures that can exhibit differential release properties were loaded with macromolecules to verify the potential of this approach for drug delivery applications. PMID- 17117839 TI - Light-induced flipping of a conserved glutamine sidechain and its orientation in the AppA BLUF domain. AB - The AppA BLUF domain is a blue light photoreceptor containing flavin. Conserved glutamine 63 is necessary for the photocycle of the protein, and its side chain has been proposed to flip in response to blue light illumination. Recently published crystal structures of AppA WT and the AppA mutant C20S describe contradictory conclusions regarding the orientation of the conserved glutamine 63 side chain in the dark. Here, we present evidence from NMR spectroscopy confirming light-induced flipping of the glutamine side chain to form a strong hydrogen bond between the glutamine 63 side chain carbonyl group and the tyrosine 21 side chain hydroxyl proton in the light-induced state. Our conclusions are consistent with published data from UV/vis absorbance and FTIR spectroscopy, as well as the crystal structure of AppA WT. PMID- 17117840 TI - Synthesis, structural characterization, and spectroscopy of the cadmium-cadmium bonded molecular species Ar'CdCdAr' (Ar' = C6H3-2,6-(C6H3-2,6-Pri2)2). AB - The synthesis and first structural characterization of a cadmium-cadmium bonded molecular compound Ar'CdCdAr' (Ar' = C6H3-2,6-(C6H3-2,6-Pri2)2) are reported. The existence of the Cd-Cd bond was established by 113Cd NMR spectroscopy and X-ray diffraction (Cd-Cd = 2.6257(5) A). Like its group 12 analogue Ar'ZnZnAr', DFT calculations showed that Ar'CdCdAr' had significant p-character in the Cd-Cd sigma-bonding HOMO. PMID- 17117841 TI - Very high third-order nonlinear optical activities of intrazeolite PbS quantum dots. AB - Zeolite-intercalated semiconductor quantum dots (QDs) have long been proposed to give very high third-order nonlinear optical (3NLO) responses. However, measurements of their 3NLO responses have not been possible due to the lack of methods to prepare optically transparent QD-incorporating zeolite films supported on optically transparent substrates and to confine QDs only within zeolite interiors. We found that the zeolite-Y films grown on indium-tin-oxide-coated glass plates (Ygs) remain firmly bonded to the substrates during ion exchange with Pb2+ ions, drying, and formation of PbS QDs by treating Pb2+ ions with H2S. A series of Ygs encapsulating different numbers (n = 0, 8, 14, 23, and 33) of PbS in a unit cell [(PbS)n-Yg] were prepared. The PbS QDs were expelled by adsorbed moisture to the external surfaces, and the expelled QDs formed large QDs. Coating of the (PbS)n-Ygs with octadecyltrimethoxysilane results in effective confinement of the QDs within the internal pores. The zeolite-encapsulated PbS QDs showed remarkably high 3NLO activities at 532 and 1064 nm which are unparalleled by other PbS QDs dispersed in other matrixes. PMID- 17117842 TI - Multimodal image-guided enzyme/prodrug cancer therapy. AB - The conjugate of bacterial cytosine deaminase (bCD) and poly-l-lysine (PLL) that was functionalized with biotin, rhodamine, and Gd3+-DOTA was synthesized and characterized. It demonstrated high relaxivity, improved enzymatic specificity to prodrug 5-fluorocytosine, low cytotoxicity, efficient cell uptake, and high enzymatic stability in fresh mouse serum and human breast cancer cell culture. PMID- 17117843 TI - Ferroelectric porous molecular crystal, [Mn3(HCOO)6](C2H5OH), exhibiting ferrimagnetic transition. AB - A porous molecular crystal with guest ethanol molecules, [Mn3(HCOO)6](C2H5OH), was found to be a new type of multifunctional molecular system, which exhibits a ferroelectric transition at 165 K and a ferrimagnetic transition at 8.5 K. [Mn3(HCOO)6](C2H5OH) will give a hint to design "multiferroic" molecular materials where ferroelectric and ferromagnetic orders coexist. PMID- 17117844 TI - Sequential hydrocarbon functionalization: allylic C-H oxidation/vinylic C-H arylation. AB - A Pd(II)/sulfoxide-catalyzed sequential allylic C-H oxidation/vinylic C-H arylation of alpha-olefins to furnish E-arylated allylic esters in high regio- and E:Z selectivities (>20:1) is reported. The broad scope of this method with respect to the alpha-olefin, carboxylic acid, and aryl boronic acid enables the rapid assembly of densely functionalized fragments for complex molecule synthesis from cheap, abundant hydrocarbon starting materials. The Pd(II)/sulfoxide catalyzed vinylic C-H arylation of electronically unbiased olefins with aryl boronic acids proceeds under oxidative, acidic conditions and mild temperatures (room temperature to 45 degrees C). PMID- 17117845 TI - Templated synthesis of single-walled carbon nanotube and metal nanoparticle assemblies in solution. AB - Carbon nanotube (CNT) and metal nanoparticle (NP) assemblies are conjugated nanosystems with potential applications in catalysis, sensing, and light harvesting. Due to poor solubility of CNTs, previously reported synthetic approaches are limited to large multi-walled CNTs, bundles of single-walled CNTs (SWNTs), or surface-bound CNTs. Here we report a solution-phase synthesis of SWNT metal NP assemblies that is generally applicable to common metal elements. Key to the process is the poly(styrene-alt-maleic acid) surfactant which disperses SWNTs in aqueous solutions and acts as templates for the binding of metal ions and metal NPs. PMID- 17117846 TI - Resonance stabilized bisdiselenazolyls as neutral radical conductors. AB - An efficient and versatile synthetic route to resonance stabilized bisselenathiazolyl and bisdiselenazolyl radicals 3 and 4 is described. Structural analysis of 3 and 4 confirm that lattice and pi-delocalization energies are sufficient to offset solid-state dimerization of the radicals and that the two selenium-containing radicals are isostructural with the all-sulfur based system 1. Variable temperature conductivity measurements indicate that sequential replacement of sulfur by selenium leads to a progressive increase in conductivity and reduction in thermal activation energy. PMID- 17117847 TI - Structure and catalytic function of Re-oxo species grafted onto H-MFI zeolite by sublimation of Re2O7. AB - Isolated Re-oxo species within spatially constrained MFI zeolite channels were synthesized via exchange with Re2O7 vapor and shown to exist as isolated Re-oxo monomers. Turnover rates and Re-oxo structures did not depend on Re content. These materials were thermally stable and catalyzed alcohol oxidation reactions at unprecedented rates and near ambient temperatures. PMID- 17117848 TI - The total synthesis of moenomycin A. AB - Moenomycin A is the only known natural antibiotic that inhibits bacterial cell wall synthesis by binding to the transglycosylases that catalyze formation of the carbohydrate chains of peptidoglycan. We report here the total synthesis of moenomycin A using the sulfoxide glycosylation method. A newly discovered byproduct of sulfoxide reactions was isolated that resulted in substantial loss of the glycosyl acceptor. A general method to suppress this byproduct was introduced, which enabled the glycosylations to proceed efficiently. The inverse addition protocol for sulfoxide glycosylations also proved essential in constructing some of the glycosidic linkages. The synthetic route is flexible and will allow for derivatives to be constructed to further analyze moenomycin A's mechanism of action. PMID- 17117849 TI - Measurement of the alpha-secondary kinetic isotope effect for the reaction catalyzed by mammalian protein farnesyltransferase. AB - Protein farnesytransferase (FTase) catalyzes the transfer of a 15-carbon prenyl group from farnesyl diphosphate (FPP) to the cysteine residue of target proteins and is a member of the newest class of zinc metalloenzymes that catalyze sulfur alkylation. Common substrates of FTase include oncogenic Ras proteins, and therefore inhibitors are under development for the treatment of various cancers. An increased understanding of the salient features of the chemical transition state of FTase may aid in the design of potent inhibitors and enhance our understanding of the mechanism of this class of zinc enzymes. To investigate the transition state of FTase we have used transient kinetics to measure the alpha secondary 3H kinetic isotope effect at the sensitive C1 position of FPP. The isotope effect for the FTase single turnover reaction using a peptide substrate that is farnesylated rapidly is near unity, indicating that a conformational change, rather than farnesylation, is the rate-limiting step. To look at the chemical step, the kinetic isotope effect was measured as 1.154 +/- 0.006 for a peptide that is farnesylated slowly, and these data suggest that FTase proceeds via a concerted mechanism with dissociative character. PMID- 17117850 TI - Chiral N-heterocyclic carbene catalyzed, enantioselective oxodiene Diels-Alder reactions with low catalyst loadings. AB - Chiral N-heterocyclic carbene (NHC) catalyzed redox reactions of racemic alpha chloroaldehydes lead to the generation of chiral enolates suitable for highly enantioselective inverse-electron-demand 1-oxodiene Diels-Alder reactions. Significantly, these reactions proceed under mild, operationally friendly reaction conditions (EtOAc, room temperature, 2-8 h) using less than 1 mol % of a chiral N-mesityl triazolium salt as the precatalyst. A broad array of densely functionalized dihydropyran-2-ones bearing either aliphatic or aromatic substiutents are formed in excellent yields and exceptional enantioselectivities from readily available reactants. Stereochemical mismatching between the racemic starting materials and the chiral catalyst is avoided by rapid epimerization of the chloroaldehydes under the reaction conditions. PMID- 17117851 TI - Gadolinium-based hybrid nanoparticles as a positive MR contrast agent. AB - A new nanoparticulate inorganic-organic hybrid-type positive contrast agent (CA), PGP/dextran-K01, was synthesized based on a GdPO4 inorganic core as a relaxation time-shortening moiety and a dextran-coating, which generates monodispersibility in water, a high relaxation-time-shortening effect by retaining a large number of water molecules in proximity of the core and toxicity reduction in in-vivo studies. This PGP/dextran-K01 nanoparticle has high r1 and r2 values and a significantly low r2/r1 value, 1.1, which is unprecedented and which is the lowest value among existing nanoparticulate CAs indicating that PGP/dextran-K01 is a positive contrast agent. Because of this low r2/r1 value and the nanoparticulate shape, PGP/dextran-K01 will be a useful clinical substitute for negative CAs based on iron oxides. PMID- 17117852 TI - Simple and versatile method for tagging phenyldiazirine photophores. AB - The first effective method for the introduction of a versatile substituent on 3 phenyl-3-trifluoromethyldiazirine has been developed. The simple preparation of a useful aldehyde intermediate allows easy access to various elaborated photoaffinity ligands, including a l-phenylalanine analog bearing a diazirine ring (TmdPhe). The asymmetric synthesis of TmdPhe was easily accomplished in gram quantities. Site-directed incorporation of this compound into the structure of a calmodulin-binding peptide using automated peptide synthesis afforded a photoreactive peptide that was successfully used for the specific labeling of calmodulin. PMID- 17117853 TI - Efficient intermolecular [2 + 2 + 2] alkyne cyclotrimerization in aqueous medium using a ruthenium(IV) precatalyst. AB - The dimeric bis(allyl)-ruthenium(IV) complex [{Ru(eta3:eta3-C10H16)(mu-Cl)Cl}2] (C10H16 = 2,7-dimethylocta-2,6-diene-1,8-diyl) was found to catalyze efficiently the [2 + 2 + 2] cyclization of terminal and internal alkynes in aqueous medium. PMID- 17117854 TI - "Click to chelate": synthesis and installation of metal chelates into biomolecules in a single step. AB - Click chemistry has been employed for the assembly of novel and efficient triazole-based multidentate chelating systems while simultaneously attaching them to molecules of biological interest. The "click-to-chelate" approach offers a powerful new tool for the modification of (bio)molecules with metal chelators for potential diagnostic and therapeutic applications. PMID- 17117855 TI - Amphiphilicity-driven organization of nanoparticles into discrete assemblies. AB - We present a method for organizing metallic nanoparticles in solution that is based on the hydrophobic effect and does not require either hydrogen bonding or molecular recognition. When amphiphilic V-shaped molecules are attached to a gold cluster, an aggregation process ensues in aqueous solution and leads to the formation of well-defined cylindrical and vesicular nanoarrays of particles. The metallic clusters densely pack at the boundary separating the hydrophobic core from the hydrophilic corona of the hybrid micelle-like aggregates. This design allows one to assemble and disassemble the nanoparticles in a reversible manner and control the size and the morphology of the arrays by changing the conditions of the solution preparation. The versatility of this method is demonstrated by its applicability to different metals with covalently attached amphiphilic arms with various chemical compositions (PS-PEO and PB-PEO) and molecular weights. PMID- 17117856 TI - Simultaneous determination of protein backbone structure and dynamics from residual dipolar couplings. AB - Determination of protein structure classically results in a single average configuration that takes no account of conformational fluctuation. Dynamics are, however, inherently linked to structure and crucial to our understanding of biological function. In this study we have used analytical descriptions of dynamic averaging of residual dipolar couplings (RDCs) to simultaneously determine the backbone structure and dynamics of protein GB3. RDCs alone are used to determine an ultrahigh-resolution structure that compares very closely with a refined X-ray structure (rmsd of 0.34 A overall backbone residues). Dynamic amplitudes reporting on motions up to the millisecond time scale reproduce the main characteristics of dynamics previously determined in conjunction with the crystal structure. The use of RDCs alone allows a bias-free comparison with a purely static approach to structure determination. Extensive cross validation clearly demonstrates that the dynamic description is superior to the static approximation. The demonstration that this level of structural resolution and dynamic detail can be extracted from RDCs supports previous indications that these parameters contain extremely precise information about biomolecular conformational sampling. PMID- 17117857 TI - Synthesis of methane in nanotube channels by a flash. AB - Inorganic synthesis of organic molecules is a significant step for the primordial life. Generally, inorganic synthesis of methane necessitates, in addition to catalyst, a high-temperature and high-pressure environment. Here we will show that such an environment could be locally and instantly realized in the channels of single-walled carbon nanotubes (SWNTs) even under room temperature and ultrahigh vacuum conditions just by a visible-light flash, owing to the ultra photothermal effect of nanomaterials. As a result, methane signals were definitely detected by using a quadrupole mass spectrometer and an optical fiber spectrometer. The mechanisms were interpreted as Fischer-Tropsch synthesis. Our results provide an alternative explanation of abiogenic methane origin. PMID- 17117858 TI - Insights into the molecular mechanism underlying polymorph selection. AB - We use molecular simulations to study polymorph selection during the crystallization of charge-stabilized colloidal suspension. By modifying the conditions of crystallization, we invert the stability of two polymorphs and induce the formation of crystallites whose structure is predominantly that of the stable polymorph. However, our simulations reveal that kinetics play a major role not only during the nucleation step but also in the growth mechanism. The growth of postcritical crystallites of the stable polymorph proceeds through a complex mechanism involving the cross-nucleation of a third metastable polymorph followed by the conversion of this third polymorph into the stable structure. PMID- 17117859 TI - Highly convergent total synthesis of (+)-acutiphycin. AB - An enantioselective, convergent, total synthesis of (+)-acutiphycin (18 steps, longest linear sequence from commercial materials) features the first application of an alkynyl ether as a macrolactone precursor in total synthesis, as well as the first example of an intermolecular, SmI2-mediated, Reformatsky fragment coupling reaction. The high convergence, efficiency, and modular nature of this synthesis make it amenable to the synthesis of structurally related compounds. PMID- 17117860 TI - X-ray crystal structures of manganese(II)-reconstituted and native toluene/o xylene monooxygenase hydroxylase reveal rotamer shifts in conserved residues and an enhanced view of the protein interior. AB - We report the X-ray crystal structures of native and manganese(II)-reconstituted toluene/o-xylene monooxygenase hydroxylase (ToMOH) from Pseudomonas stutzeri OX1 to 1.85 and 2.20 A resolution, respectively. The structures reveal that reduction of the dimetallic active site is accompanied by a carboxylate shift and alteration of the coordination environment for dioxygen binding and activation. A rotamer shift in a strategically placed asparagine 202 accompanies dimetallic center reduction and is proposed to influence protein component interactions. This rotamer shift is conserved between ToMOH and the corresponding residue in methane monooxygenase hydroxylase (MMOH). Previously unidentified hydrophobic pockets similar to those present in MMOH are assigned. PMID- 17117861 TI - Low polydispersity star polymers via cross-linking macromonomers by ATRP. AB - A new method for the synthesis of star polymers with high molecular weight and narrow molecular weight distribution (MWD) is presented. The star copolymers were prepared by atom transfer radical (co)polymerization (ATRP) of linear macromonomers with divinyl cross-linkers using low molar mass initiator. In contrast to prior "arm-first" procedures with macroinitiators, by using lower ratio of concentration of initiator to MM, the star polymers contained less initiating sites in the core, which decreased the extent of star-star coupling reactions and resulted in formation of star polymers with narrower MWD. Addition of more cross-linker and initiator during the reaction increased the star molecular weight and star yield while retaining the narrow MWD of the star polymer. For example, we synthesized a star polymer with molecular weight Mn = 466 000 g/mol and Mw/Mn < 1.2 in >98% yield. PMID- 17117862 TI - Antibodies with broad specificity to azaspiracids by use of synthetic haptens. AB - The development of general, sensitive, portable, and quantitative assays for the azaspiracid (AZA) class of marine toxins is urgently needed. Use of a synthetic hapten containing rings F-I of AZA to generate antibodies that cross-react with the AZAs via their common C28-C40 domain and use of these antibodies in ELISA and immunoaffinity columns are reported. This approach has many advantages over using intact azaspiracids (AZAs) derived from environmental samples or total synthesis as haptens for antibody development. A derivative of the levorotatory C28-C40 azaspiracid domain (1) was synthesized efficiently using a one-pot Staudinger reduction/intramolecular aza-Wittig reaction-imine capture sequence to form the H I ring spiroaminal and a double intramolecluar hetero-Michael addition to assemble the F-G ring ketal. Conjugation of the hapten 1 to cBSA and immunization in sheep generated antibodies that recognized and bound to ovalbumin-conjugated 1 in the absence of AZA1. This binding was inhibited by 1 in a concentration dependent manner. A mixture of AZA1, AZA2, AZA3, and AZA6 caused a degree of inhibition of antibody binding consistent with its total AZA content, rather than just its content of AZA1. This result suggests that the antibodies also have a similar affinity for AZA2, AZA3, and AZA6 as they do for AZA1 and that such antibodies are suitable for analysis of AZAs in shellfish samples. PMID- 17117863 TI - Origin of the linear relationship between CH2/NH/O-SWNT reaction energies and sidewall curvature: armchair nanotubes. AB - The origin of the linear relationship between the reaction energy of the CH2/NH/O exo and endo additions to armchair (n, n) single-walled carbon nanotubes (SWNTs) and the inverse tube diameter (1/d) measuring sidewall curvature was elucidated using density functional theory and density functional tight binding methods for finite-size SWNT models with n = 3, 4, ..., 13. A nearly perfect linear relationship between DeltaE and 1/d all through exohedral (positive curvature) and endohedral (negative curvature) additions is due to cancellation between the quadratic contributions of the SWNT deformation energy and the interaction energy (INT) between the deformed SWNT and CH2/NH/O adducts. Energy decomposition analysis shows that the quadratic contributions in electrostatic, exchange, and orbital terms mostly cancel each other, making INT weakly quadratic, and that the linear 1/d dependence of INT, and therefore of DeltaE, is a reflection of the 1/d dependence of the back-donative orbital interaction of b1 symmetry from the occupied CH2/NH/O p pi orbital to the vacant C=C pi* LUMO of the SWNT. We also discuss the origin of the two isomers (open and three-membered ring) of the exohedral addition product and explain the behavior of their associated minima on the C-C potential energy surfaces with changing d. PMID- 17117864 TI - Geometry, energetics, and dynamics of hydrogen bonds in proteins: structural information derived from NMR scalar couplings. AB - An accurate description of hydrogen bonds is essential to identify the determinants of protein stability and function as well as folding and misfolding behavior. We describe a method of using J couplings through hydrogen bonds as ensemble-averaged restraints in molecular dynamics simulations. Applications to the cases of ubiquitin and protein G show that these scalar couplings provide powerful structural information that, when used through the methodology that we present here, enables the description of the geometry and energetics of hydrogen bonds with an accuracy approaching that of high-resolution X-ray structures. PMID- 17117865 TI - Neutron diffraction and simulation studies of CsNO3 and Cs2CO3 solutions. AB - Neutron diffraction with isotopic substitution (NDIS) experiments and molecular dynamics (MD) simulations have been used to study the structuring in aqueous solution of two cesium salts, cesium carbonate, and cesium nitrate. As was previously found for guanidinium salts of carbonate, mesoscopic-scale clusters were seen to form in the Cs2CO3 solution both in the MD simulations and in the diffraction experiments. No such large scale ion clusters were found in the CsNO3 solutions in either the modeling or experiments. The results are dominated by the strength and geometry of the direct first-neighbor interactions, which explain the differences in the clustering behavior between the two solutions without need to refer to longer-range water-water structuring. PMID- 17117866 TI - Supramolecular bis(rutheniumphthalocyanine)-Perylenediimide ensembles: simple complexation as a powerful tool toward long-lived radical ion pair states. AB - A novel supramolecular electron donor-acceptor hybrid (1) has been designed through axial coordination of a perylenebisimide moiety [BPyPDI], bearing two 4 pyridyl substituents at the imido positions, to the ruthenium(II) metal centers of two phthalocyanines [Ru(CO)Pc]. This modular protocol enables access to electron donor-acceptor hybrids with potentially great design flexibility. The new array (1) has been characterized by standard spectroscopic methods, and its photophysical behavior has been established by using ultrafast and fast time resolved techniques. Photoexcitation of either chromophore leads to a product that is essentially identical for both pathways, that is, evolving from the [Ru(CO)Pc] or [BPyPDI] singlet excited state. Features of the photoproduct are new transient maxima at 530 and 725 nm, plus transient minima at 580 nm and 650 nm. Based on the radiolytically generated [BPyPDI*-] (i.e., one-electron reduction of [BPyPDI]) and [Ru(CO)Pc*+] (i.e., one-electron oxidation of [Ru(CO)Pc]) features, which in the 300 and 900 nm range remarkably resemble those noted for photoexcited 1, we attribute the photolytically generated species to the composite spectrum of the [Ru(CO)Pc*+ -BPyPDI*- -RuCOPc] radical ion pair state. Its lifetime, which is on the order of 115 +/- 5 ns, reveals a significant stabilization and confirms that the strongly exothermic charge recombination dynamics are placed deeply in the inverted region of the Marcus parabola. PMID- 17117867 TI - Hydration of mononucleotides. AB - The sequential addition of water molecules to protonated and deprotonated forms of the four mononucleotides dAMP, dCMP, dGMP, and dTMP was studied experimentally by equilibrium measurements using an electrospray mass spectrometer equipped with a drift cell and theoretically by computational methods including molecular modeling and density functional theory calculations. Experiments were carried out in positive and negative ion mode, and calculations included the protonated and deprotonated forms of the four nucleotides. For deprotonated anionic nucleotides the experimental enthalpies of hydration (DeltaH degrees n) were found to be similar for all four systems and varied between -10.1 and -11.5 kcal mol-1 for the first water molecule (n = 1) and -8.3 and -9.6 kcal mol-1 for additional water molecules (n = 2-4). Theory indicated that the first water molecule binds to the charge-carrying phosphate group. Simulations of deprotonated mononucleotides with four water molecules yielded a large number of structures with similar energies. In some of the structures all four water molecules cluster around the phosphate group, and in other structures the four water molecules each hydrate a different functional group of the nucleotide. These include the phosphate group, the deoxyribose hydroxyl group, and various functional groups on the nucleobases. Experimental DeltaH degrees 1 values for the protonated cationic mononucleotides ranged from -10.5 to -13.5 kcal mol-1 with more negative values (< or =-12 kcal mol-1) for dCMP, dGMP, and dTMP and the least negative value for dAMP. For n = 2-4 DeltaH degrees n values varied from -6.9 to -9.7 kcal/mol and were similar in value to the deprotonated nucleotides except for dAMP. Theory on the protonated nucleotides indicated that the first water molecule binds to the charge-carrying group for dCMP, dGMP, and dTMP. For protonated dAMP, on the other hand, the charge-carrying N3 group is well self-solvated by the phosphate group and not readily available for a hydrogen bond with the water molecule. The insight gained on nucleotide stabilization by individual water molecules is used to discuss the competition between hydration of individual nucleotides and Watson Crick base pairing. PMID- 17117868 TI - N-methylated cyclic pentaalanine peptides as template structures. AB - The N-methylation of cyclic peptides can be used to modify the activity and/or selectivity of biologically active peptides. As N-methylation introduces different flexibility and lipophilicity, it can also improve the bioavailability (the ADMET profile). To search for conformationally constrained cyclic peptides, a library of 30 different N-methylated peptides with the basic sequence cyclo(-D Ala-L-Ala4-) was synthesized. Based on the NMR analysis, seven of these peptides exhibited single conformations (>98%). The structural features of these peptides were determined by a combination of NMR and distance geometry and then further refined by molecular dynamics simulations in an explicit DMSO solvent box. The structures provided from these efforts can now serve as templates for the rational design of cyclic pentapeptides with a distinct backbone conformation or for "spatial screening" to explore the bioactive conformation of medically important peptide systems. PMID- 17117869 TI - Conformationally constrained 2'-N,4'-C-ethylene-bridged thymidine (aza-ENA-T): synthesis, structure, physical, and biochemical studies of aza-ENA-T-modified oligonucleotides. AB - The 2'-deoxy-2'-N,4'-C-ethylene-bridged thymidine (aza-ENA-T) has been synthesized using a key cyclization step involving 2'-ara-trifluoromethylsufonyl 4'-cyanomethylene 11 to give a pair of 3',5'-bis-OBn-protected diastereomerically pure aza-ENA-Ts (12a and 12b) with the fused piperidino skeleton in the chair conformation, whereas the pentofuranosyl moiety is locked in the North-type conformation (7 degrees < P < 27 degrees, 44 degrees < phi m < 52 degrees). The origin of the chirality of two diastereomerically pure aza-ENA-Ts was found to be due to the endocyclic chiral 2'-nitrogen, which has axial N-H in 12b and equatorial N-H in 12a. The latter is thermodynamically preferred, while the former is kinetically preferred with Ea = 25.4 kcal mol-1, which is thus far the highest observed inversion barrier at pyramidal N-H in the bicyclic amines. The 5'-O-DMTr-aza-ENA-T-3'-phosphoramidite was employed for solid-phase synthesis to give four different singly modified 15-mer antisense oligonucleotides (AONs). Their AON/RNA duplexes showed a Tm increase of 2.5-4 degrees C per modification, depending upon the modification site in the AON. The relative rates of the RNase H1 cleavage of the aza-ENA-T-modified AON/RNA heteroduplexes were very comparable to that of the native counterpart, but the RNA cleavage sites of the modified AON/RNA were found to be very different. The aza-ENA-T modifications also made the AONs very resistant to 3' degradation (stable over 48 h) in the blood serum compared to the unmodified AON (fully degraded in 4 h). Thus, the aza-ENA-T modification in the AON fulfilled three important antisense criteria, compared to the native: (i) improved RNA target affinity, (ii) comparable RNase H cleavage rate, and (iii) higher blood serum stability. PMID- 17117870 TI - Unraveling the aflatoxin-FAPY conundrum: structural basis for differential replicative processing of isomeric forms of the formamidopyrimidine-type DNA adduct of aflatoxin B1. AB - Aflatoxin B1 (AFB) epoxide forms an unstable N7 guanine adduct in DNA. The adduct undergoes base-catalyzed ring opening to give a highly persistent formamidopyrimidine (FAPY) adduct which exists as a mixture of forms. Acid hydrolysis of the FAPY adduct gives the FAPY base which exists in two separable but interconvertible forms that have been assigned by various workers as functional, positional, or conformational isomers. Recently, this structural question became important when one of the two major FAPY species in DNA was found to be potently mutagenic and the other a block to replication [Smela, M. E.; Hamm, M. L.; Henderson, P. T.; Harris, C. M.; Harris, T. M.; Essigmann, J. M. Proc. Natl. Acad. Sci. U.S.A. 2002, 99, 6655-6660]. NMR studies carried out on the AFB-FAPY bases and deoxynucleoside 3',5'-dibutyrates now establish that the separable FAPY bases and nucleosides are diastereomeric N5 formyl derivatives involving axial asymmetry around the congested pyrimidine C5-N5 bond. Anomerization of the protected beta-deoxyriboside was not observed, but in the absence of acyl protection, both anomerization and furanosyl --> pyranosyl ring expansion occurred. In oligodeoxynucleotides, two equilibrating FAPY species, separable by HPLC, are assigned as anomers. The form normally present in duplex DNA is the mutagenic species. It has previously been assigned as the beta anomer by NMR (Mao, H.; Deng, Z. W.; Wang, F.; Harris, T. M.; Stone, M. P. Biochemistry 1998, 37, 4374-4387). In single-stranded environments the dominant species is the beta anomer; it is a block to replication. PMID- 17117871 TI - 15N solid-state NMR provides a sensitive probe of oxidized flavin reactive sites. AB - Flavins are central to the reactivity of a wide variety of enzymes and electron transport proteins. There is great interest in understanding the basis for the different reactivities displayed by flavins in different protein contexts. We propose solid-state nuclear magnetic resonance (SS-NMR) as a tool for directly observing reactive positions of the flavin ring and thereby obtaining information on their frontier orbitals. We now report the SS-NMR signals of the redox-active nitrogens N1 and N5, as well as that of N3. The chemical shift tensor of N5 is over 720 ppm wide, in accordance with the predictions of theory and our calculations. The signal of N3 can be distinguished on the basis of coupling to 1H absent for N1 and N5, as well as the shift tensor span of only 170 ppm, consistent with N3's lower aromaticity and lack of a nonbonding lone pair. The isotropic shifts and spans of N5 and N1 reflect two opposite extremes of the chemical shift range for "pyridine-type" N's, consistent with their electrophilic and nucleophilic chemical reactivities, respectively. Upon flavin reduction, N5's chemical shift tensor contracts dramatically to a span of less than 110 ppm, and the isotropic chemical shift changes by approximately 300 ppm. Both are consistent with loss of N5's nonbonding lone pair and decreased aromaticity, and illustrate the responsiveness of the 15N chemical shift principal values to electronic structure. Thus. 15N chemical shift principal values promise to be valuable tools for understanding electronic differences that underlie variations in flavin reactivity, as well as the reactivities of other heterocyclic cofactors. PMID- 17117872 TI - Direct observation of substrate-enzyme complexation by surface forces measurement. AB - The substrate-enzyme complexation of heptaprenyl diphosphate synthase was directly investigated using colloidal probe atomic force microscopy (AFM) and a quartz crystal microbalance (QCM) in order to obtain new insights into the molecular mechanism of the enzyme reaction. This enzyme is composed of two dissociable subunits that exhibit a catalytic activity only when they are associated together in the presence of a cofactor, Mg2+, and a substrate, farnesyl diphosphate (FPP). The QCM measurement revealed that FPP was preferentially bound to subunit II in the presence of Mg2+, while the AFM measurement showed that the adhesive force between the subunits was observed only in the presence of both Mg2+ and FPP. This is the first direct demonstration of the specific interaction involved in the enzyme reaction. The dependence of the Mg2+ concentration on the specific interaction between subunits I and II well agreed with that on the enzyme activity of heptaprenyl diphosphate synthase. This indicated that the observed adhesive forces were indeed involved in the catalytic reaction of this enzyme. On the basis of these results, we discussed the processes involved in the substrate-enzyme complexation. The first, the substrate FPP bound to subunit II using Mg2+, followed by the formation of the subunit I FPP-Mg2+-subunit II complex. Our study showed a very useful methodology for examining the elemental processes of biological reactions such as an enzyme reaction. PMID- 17117873 TI - Enantioselective synthesis of diversely substituted quaternary 1,4-benzodiazepin 2-ones and 1,4-benzodiazepine-2,5-diones. AB - Benzodiazepines are privileged scaffolds in medicinal chemistry, but enantiopure examples containing quaternary stereogenic centers are extremely rare. We demonstrate that installation of the di(p-anisyl)methyl (DAM) group at N1 of 1,4 benzodiazepin-2-ones and 1,4-benzodiazepine-2,5-diones derived from enantiopure proteinogenic amino acids allows retentive replacement of the C3-proton via a deprotonation/trapping protocol. A wide variety of carbon and nitrogen electrophiles function well in this reaction, providing the corresponding quaternary benzodiazepines with excellent enantioselectivity. Deprotonation/trapping experiments on a pair of diastereomeric 1,4-benzodiazepine 2,5-diones provide evidence for a key role of conformational chirality in these enantioselective reactions. Acidic removal of the DAM group is fast and high yielding and can be performed selectively in the presence of a N-Boc indole. Thus the synthesis of quaternary benzodiazepines with diverse N1 functionality can now be accomplished. PMID- 17117874 TI - Lipid lateral mobility and membrane phase structure modulation by protein binding. AB - Using a combination of fluorescence correlation and infrared absorption spectroscopies, we characterize lipid lateral diffusion and membrane phase structure as a function of protein binding to the membrane surface. In a supported membrane configuration, cholera toxin binding to the pentasaccharaide headgroup of membrane-incorporated GM1 lipid alters the long-range lateral diffusion of fluorescently labeled probe lipids, which are not involved in the binding interaction. This effect is prominently amplified near the gel-fluid transition temperature, Tm, of the majority lipid component. At temperatures near Tm, large changes in probe lipid diffusion are measured at average protein coverage densities as low as 0.02 area fraction. Spectral shifts of the methylene symmetric and asymmetric stretching modes in the lipid acyl chain confirm that protein binding alters the fraction of lipid in the gel phase. PMID- 17117875 TI - Chemistry of periodate-mediated cross-linking of 3,4-dihydroxylphenylalanine containing molecules to proteins. AB - Chemical cross-linking is an attractive approach to map peptide-protein and protein-protein complexes. Previously, we explored 3,4-dihydroxylphenylalanine (DOPA) as a protein cross-linking agent upon periodate oxidation (Burdine, L.; Gillette, T. G.; Lin, H.-J.; Kodadek, T. J. Am. Chem. Soc. 2004, 126, 11442 11443). We report here a study on the chemistry of DOPA-protein cross-linking. First, using a peptide nucleic acid templated system, we identified the alpha amino, epsilon-amino of Lys, imidazole of His, and thiol of Cys as functional groups capable of attacking DOPA ortho-quinone. Second, we demonstrated that periodate-induced DOPA-protein cross-linking could be carried out efficiently at neutral pH in the presence of excess aliphatic 1,2-diols such as ethylene glycol, lactose, and adenosine triphosphate. This result indicated that DOPA-protein cross-linking and 1,2-diol oxidative cleavage proceed via different mechanisms and that carbohydrates will not interfere with this process when carried out in crude cell extracts or on intact cells. PMID- 17117876 TI - Intramolecular sulfur transfer in N-enoyl oxazolidine-2-thiones promoted by Bronsted acids. Practical asymmetric synthesis of beta-mercapto carboxylic acids and mechanistic insights. AB - The ability of Bronsted acids alone to efficiently promote the sulfur transfer process in N-enoyl oxazolidine-2-thiones to give beta-mercapto carbonyl derivatives is demonstrated. The reactions proceed with essentially perfect diastereocontrol for a range of alkyl-substituted N-enoyl oxazolidine-2-thiones (d.r. regularly above 98:2) and high selectivity for most aryl-substituted counterparts (d.r. typically above 92:8). Importantly, the reaction works remarkably well in beta,beta-disubstituted N-enoyl oxazolidine-2-thiones as well, giving rise to quaternary C-S stereocenters in selectivities usually above 95:5. The relative efficiency of a range of acids (trifluoroacetic, difluoroacetic, acetic, triflic) is assessed showing TFA and TfOH as the most efficient and acetic acid as a totally inefficient reaction promoter. The new procedure complements the Lewis acid promoted reaction previously described by our group in two aspects: First, stereodivergent results are obtained for the Lewis acid or Bronsted acid promoted reactions of beta,beta-disubstituted enoyl compounds. Second, while the Bronsted acid promoted reactions are stereospecific, providing a good correlation between the substrate E/Z configuration and products stereochemistry, the reactions mediated by Lewis acids (BF3/OEt2) provide invariant d.r. values regardless of the E/Z composition of the starting olefin. The synthetic value of the method is illustrated by (a) removal of the oxazolidinone moiety from the rearranged products under reducing conditions (NaBH4, H2O-THF) which yields beta-mercapto alcohols and (b) treatment with Sm(OTf)3 in MeOH which affords the corresponding beta-mercapto carboxylic esters, both categories of compounds being isolated in up to 97% ee. Remarkably, the method constitutes the first general approach to highly enantioenriched building blocks bearing a quaternary C-S stereocenter. On the other hand, spectroscopic and inhibition experiments are carried out that demonstrate the participation of protons also in the Lewis acid promoted reactions. Finally, the computational studies carried out at the B3LYP/6-31G* level give support for an activation of the substrate enoyl by complexation with two molecules of either the Bronsted or Lewis acid and serve to explain the stereochemical outcome of the reactions. PMID- 17117877 TI - Dissection of aminoglycoside-enzyme interactions: a calorimetric and NMR study of neomycin B binding to the aminoglycoside phosphotransferase(3')-IIIa. AB - In this work, for the first time, we report pKa values of the amino functions in a target-bound aminoglycoside antibiotic, which permitted dissection of the thermodynamic properties of an enzyme-aminoglycoside complex. Uniformly enriched 15N-neomycin was isolated from cultures of Streptomyces fradiae and used to study its binding to the aminoglycoside phosphotransferase(3')-IIIa (APH) by 15N NMR spectroscopy. 15N NMR studies showed that binding of neomycin to APH causes upshifts of approximately 1 pKa unit for the amines N2' and N2' '' while N6' experienced a 0.3 pKa unit shift. The pKa of N6' '' remained unaltered, and resonances of N1 and N3 showed significant broadening upon binding to the enzyme. The binding-linked protonation and pH dependence of the association constant (Kb) for the enzyme-aminoglycoside complex was determined by isothermal titration calorimetry. The enthalpy of binding became more favorable (negative) with increasing pH. At high pH, binding-linked protonation was attributable mostly to the amino functions of neomycin; however, at neutral pH, functional groups of the enzyme, possibly remote from the active site, also underwent protonation/deprotonation upon formation of the binary enzyme-neomycin complex. The Kb for the enzyme-neomycin complex showed a complicated dependence on pH, indicating that multiple interactions may affect the affinity of the ligand to the enzyme and altered conditions, such as pH, may favor one or another. This work highlights the importance of determining thermodynamic parameters of aminoglycoside-target interactions under different conditions before making attributions to specific sites and their effects on these global parameters. PMID- 17117878 TI - Topological control in heterometallic metal-organic frameworks by anion templating and metalloligand design. AB - Several new heterometallic metal-organic frameworks (MOFs) based on tris(dipyrrinato) metalloligands and Ag+ salts are reported. MOFs were prepared systematically to examine the effects of the core metal ion, counteranion, and ligand structure on the topology of the resultant network. The effect of the metal ion (Fe3+ vs Co3+) on MOF structure was generally found to be negligible, thereby permitting the facile synthesis of trimetallic Fe/Co/Ag networks. The choice of anion (e.g., silver salt) was found to have a pronounced effect on the MOF topology. Networks prepared with salts of AgO3SCF3 and AgBF4 reliably formed three-dimensional (10,3) nets, whereas use of AgPF6 and AgSbF6 produced two dimensional (6,3) honeycomb nets. The topology generated upon formation of the MOF was found to be robust in certain cases, as demonstrated by anion-exchange experiments. Anion exchange was confirmed by X-ray crystallography in a rare set of apparent single-crystal-to-single-crystal transformations. The data presented here strongly suggest that the coordinative ability of the anion does not play a significant role in the observed templating effect. Finally, changes in the length of the tris(dipyrrinato) metalloligand were found to override the anion templating effect, resulting exclusively in two-dimensional (6,3) nets. These studies provide a basis for the rational design of MOF topologies by choice of ligand structure and anion templating effects. Furthermore, the results demonstrate the ability of carefully designed metalloligands to generate MOFs of structure strikingly similar to that of their organic counterparts. PMID- 17117879 TI - Cation exchange in lipophilic G-quadruplexes: not all ion binding sites are equal. AB - Lipophilic guanosine derivatives that form G-quadruplexes are promising building blocks for ionophores and ion channels. Herein, cation exchange between solvated cations (K+ and NH4+) and bound cations in the G-quadruplex [G1]16.4Na+.4DNP- was studied by electrospray ionization mass spectrometry and solution 1H, 15N NMR spectroscopy. The ESI-MS and 1H NMR data provided evidence for the formation of mixed-cationic Na+, K+ G-quadruplexes. The use of 15NH4+ cations in NMR titrations, along with 15N-filtered 1H NMR and selective NOE experiments, identified two mixed-cationic intermediates in the cation exchange pathway from [G1]16.4Na+.4DNP- to [G1]16.4NH4+.4DNP-. The central Na+, bound between the two symmetry-related G8-Na+ octamers, exchanges with either K+ or NH4+ before the two outer Na+ ions situated within the C4 symmetric G8 octamers. A structural rationale, based on differences in the cations' octahedral coordination geometries, is proposed to explain the differences in site exchange for these lipophilic G-quadruplexes. Large cations such as Cs+ can be exchanged into the central cation binding site that holds the two symmetry-related C4 symmetric G8 octamer units together. The potential relevance of these findings to both supramolecular chemistry and DNA G-quadruplex structure are discussed. PMID- 17117880 TI - A rational approach to minimal high-resolution cross-reactive arrays. AB - We report a rational approach to the construction of cross-reactive arrays for steroids consisting of five to seven sensors incorporating modified oligonucleotides. The sensors for our arrays were selected to maximize their differential responses to the two steroids most different in an arbitrarily chosen parameter named "shape-length". The arrays incorporated three previously unreported types of sensors identified through a massive screening effort: (1) three-way junction sensors with neutralized charges within junction; (2) "self aggregating sensors"; and (3) sensors incorporating fluorophore directly in a three-way junction as a spacer. The arrays were tested on seven steroids and an alkaloid (cocaine) over a range of concentrations, and achieved 92-96% accuracy in class assignments, despite the close structural similarities between steroids. PMID- 17117881 TI - Simulating self-assembly of ZnS nanoparticles into mesoporous materials. AB - Characterization of materials is crucial for the quantification and prediction of their physical, chemical, and mechanical properties. However, as the complexity of a system increases, so do the challenges involved in elucidating its structure. While molecular simulation and modeling have proved invaluable as complements to experiment, such simulations now face serious challenges: new materials are being synthesized with ever increasing structural complexity, and it may soon prove impossible to generate models that are sufficiently realistic to describe them adequately. Perhaps, ultimately, it will only be possible to generate such models by simulating the synthetic process itself. Here, we attempt such a strategy to generate full atomistic models for mesoporous molecular sieves. As in experiment, this is done by allowing nanoparticles to self-assemble at high temperature to form an amorphous mesoporous framework. The temperature is then reduced, and the system is allowed to crystallize. Animations of atomic trajectories, available as Supporting Information, reveal the evolution of multiple seeds which propagate to form a complex framework. The products are polycrystalline mesoporous framework structures containing cavities connected by channels running along "zero", one, two, and three perpendicular directions. We suggest that it is easier to generate these model structures by attempting to simulate the synthetic process rather than by using more conventional techniques. The strategy is illustrated using ZnS as a model system. Further development of the mathematics of minimal surfaces will advance our understanding of these structures. PMID- 17117882 TI - FAST-NMR: functional annotation screening technology using NMR spectroscopy. AB - An abundance of protein structures emerging from structural genomics and the Protein Structure Initiative (PSI) are not amenable to ready functional assignment because of a lack of sequence and structural homology to proteins of known function. We describe a high-throughput NMR methodology (FAST-NMR) to annotate the biological function of novel proteins through the structural and sequence analysis of protein-ligand interactions. This is based on basic tenets of biochemistry where proteins with similar functions will have similar active sites and exhibit similar ligand binding interactions, despite global differences in sequence and structure. Protein-ligand interactions are determined through a tiered NMR screen using a library composed of compounds with known biological activity. A rapid co-structure is determined by combining the experimental identification of the ligand binding site from NMR chemical shift perturbations with the protein-ligand docking program AutoDock. Our CPASS (Comparison of Protein Active Site Structures) software and database are then used to compare this active site with proteins of known function. The methodology is demonstrated using unannotated protein SAV1430 from Staphylococcus aureus. PMID- 17117883 TI - Determination of the adsorption isotherm of methanol on the surface of ice. An experimental and grand canonical Monte Carlo simulation study. AB - The adsorption isotherm of methanol on ice at 200 K has been determined both experimentally and by using the Grand Canonical Monte Carlo computer simulation method. The experimental and simulated isotherms agree well with each other; their deviations can be explained by a small (about 5 K) temperature shift in the simulation data and, possibly, by the non-ideality of the ice surface in the experimental situation. The analysis of the results has revealed that the saturated adsorption layer is monomolecular. At low surface coverage, the adsorption is driven by the methanol-ice interaction; however, at full coverage, methanol-methanol interactions become equally important. Under these conditions, about half of the adsorbed methanol molecules have one hydrogen-bonded water neighbor, and the other half have two hydrogen-bonded water neighbors. The vast majority of the methanols have a hydrogen-bonded methanol neighbor, as well. PMID- 17117884 TI - On the mechanism of hydrolysis of phosphate monoesters dianions in solutions and proteins. AB - The nature of the hydrolysis of phosphate monoester dianions in solutions and in proteins is a problem of significant current interest. The present work explores this problem by systematic calculations of the potential surfaces of the reactions of a series of phosphate monoesters with different leaving groups. These calculations involve computational studies ranging from ab initio calculations with implicit solvent models to ab initio QM/MM free energy calculations. The calculations reproduce the observed linear free energy relationship (LFER) for the solution reaction and thus are consistent with the overall experimental trend and can be used to explore the nature of the transition state (TS) region, which is not accessible to direct experimental studies. It is found that the potential surface for the associative and dissociative paths is very flat and that the relative height of the associative and dissociative TS is different in different systems. In general, the character of the TS changes from associative to dissociative upon decrease in the pKa of the leaving group. It is also demonstrated that traditional experimental markers such as isotope effects and the LFER slope cannot be used in a conclusive way to distinguish between the two classes of transition states. In addition it is found that the effective charges of the TS do not follow the previously assumed simple rule. Armed with that experience we explore the free energy surface for the GTPase reaction of the RasGap system. In this case it is found that the surface is flat but that the lowest TS is associative. The present study indicates that the nature of the potential surfaces for the phosphoryl transfer reactions in solution and proteins is quite complicated and cannot be determined in a conclusive way without the use of careful theoretical studies that should, of course, reproduce the available experimental information. PMID- 17117885 TI - Solution-phase single quantum dot fluorescence resonance energy transfer. AB - We present a single particle fluorescence resonance energy transfer (spFRET) study of freely diffusing self-assembled quantum dot (QD) bioconjugate sensors, composed of CdSe-ZnS core-shell QD donors surrounded by dye-labeled protein acceptors. We first show that there is direct correlation between single particle and ensemble FRET measurements in terms of derived FRET efficiencies and donor acceptor separation distances. We also find that, in addition to increased sensitivity, spFRET provides information about FRET efficiency distributions which can be used to resolve distinct sensor subpopulations. We use this capacity to gain information about the distribution in the valence of self-assembled QD protein conjugates and show that this distribution follows Poisson statistics. We then apply spFRET to characterize heterogeneity in single sensor interactions with the substrate/target and show that such heterogeneity varies with the target concentration. The binding constant derived from spFRET is consistent with ensemble measurements. PMID- 17117886 TI - Temperature-responsive protein pores. AB - We describe temperature-responsive protein pores containing single elastin-like polypeptide (ELP) loops. The ELP loops were placed within the cavity of the lumen of the alpha-hemolysin (alphaHL) pore, a heptamer of known crystal structure. The cavity is roughly spherical with a molecular surface volume of about 39,500 A3. In an applied potential, the wild-type alphaHL pore remained open for long periods. In contrast, the ELP loop-containing alphaHL pores exhibited transient current blockades, the nature of which depended on the length and sequence of the inserted loop. Together with similar results obtained with poly(ethylene glycols) covalently attached within the cavity, the data suggest that the transient current blockades are caused by excursions of ELP into the transmembrane beta barrel domain of the pore. Below its transition temperature, the ELP loop is fully expanded and blocks the pore completely, but reversibly. Above its transition temperature, the ELP is dehydrated and the structure collapses, enabling a substantial flow of ions. Potential applications of temperature responsive protein pores in medical biotechnology are discussed. PMID- 17117887 TI - Distinct kinetic pathways generate organogel networks with contrasting fractality and thixotropic properties. AB - The kinetics of the isothermal transformation of sols, comprised of a low molecular-mass organogelator (LMOG) and an organic liquid, to their organogel phases have been followed by circular dichroism (CD), fluorescence, small angle neutron scattering (SANS), and rheological methods. The thixotropic properties (in the sense that severe shearing followed by rest lead to reestablishment of viscoelasticity) of the gels have been examined as well by rheological measurements. The compositions of the samples were either 5alpha-cholestan-3beta yl N-(2-naphthyl) carbamate (CNC) in an n-alkane (n-octane or n-dodecane) or 3beta-cholesteryl N-(2-naphthyl) carbamate (CeNC) in ethyl acetate. Values of Df, the mass fractal dimension of the microcrystalline self-assembled fibrillar networks (SAFINs) in the gels, have been extracted from the kinetic data using a model developed by Dickinson (J. Chem. Soc., Faraday Trans. 1997, 93, 111). The Df values, 1.1-1.3 for the CeNC gels and 1.3-1.4 or 1.6-1.8 (depending on the temperature of incubation of the sol phase) for CNC gels, are consistent with the gel network structures observed by optical microscopy. In addition, comparison of the temperature dependence of both n (the Avrami component) and K (the Avrami "rate constant") for CeNC/ethyl acetate gelation with those reported previously for gelation of CNC/n-alkane sols demonstrate that the very small change of a single bond in CNC to a double bond in CeNC causes significant differences in their gelation abilities and gel properties. The rheological measurements on CNC/n-alkane gels with spherulitic SAFIN units, formed by incubation of their sols at < or =28 degrees C, indicate that they are thixotropic. Gels with the same chemical composition but formed by incubation of their sols at > or =30 degrees C, leading to fiberlike SAFIN units, remain liquidlike after shearing regardless of the periods they are at rest. The time-dependent viscoelastic properties of the gel networks are treated according to a stretched exponential model. The observations from these studies provide detailed insights into the mechanisms of formation of molecular organogel phases and demonstrate the extreme sensitivity of the SAFINs and viscoelastic properties of such organogels to slight modifications in LMOG structure or sample history. PMID- 17117888 TI - Laparoscopic photodynamic diagnosis of ovarian cancer peritoneal micro metastasis: an experimental study. AB - The goal of this study was to assess the interest of photodynamic diagnosis (PDD) for laparoscopic detection of peritoneal micro metastasis in ovarian carcinoma. Using an experimental animal model, intraperitoneal injection of aminolevulinic acid (ALA) and hexylester of aminolevulinic acid (He-ALA) were compared in order to improve laparoscopic detection of ovarian peritoneal carcinomatosis. Twenty one 344 Fischer female rats received an intra peritoneal injection of 106 NuTu-19 cells. At day 22, carcinomatosis with micro peritoneal metastasis was obtained. Rats were randomized in three groups concerning intra peritoneal injection before laparoscopic staging: 5-ALA hydrochloride, HE-ALA and sterile water. Using D Light system, laparoscopic peritoneal exploration was performed with white light (WL) first and then with blue light (BL). The main objective was to assess feasibility and sensibility of laparoscopic PDD for nonvisible peritoneal micro metastasis of ovarian cancer. The main parameter was the confirmation of neoplasic status of fluorescent foci by histology. Concerning PDD after intraperitoneal injection of 5-ALA, mean values of lesions seen is higher than without fluorescence (32 vs 20.7; P = 0.01). Using He-ALA, mean values of detected lesions is higher than without fluorescence (42.9 vs 33.6; P < 0.001). Neoplasic status of fluorescent foci was confirmed in 92.8% of cases (39/42). Using 5-ALA, fluorescence of cancerous tissue is significantly higher than that of normal tissue in all the rats (ratio 1.17) (P = 0.01). With He-ALA, intensity of fluorescence is significantly higher in cancerous tissue compared to normal tissue, irrespective of the rat studied (ratio 1.22; P < 0.001). PMID- 17117889 TI - Picosecond multidimensional fluorescence spectroscopy: a tool to measure real time protein dynamics during function. AB - Advanced multidimensional time-correlated single photon counting (mdTCSPC) and picosecond time-resolved fluorescence in combination with site-directed fluorescence labeling are valuable tools to study the properties of membrane protein surface segments on the pico- to nanoseconds time scale. Time-resolved fluorescence anisotropy changes of protein bound fluorescent probes reveal changes in protein dynamics and steric restriction. In addition, the change in fluorescence lifetime and intensity of the covalently bound fluorescent dye is indicative of environmental changes at the protein surface. In this study, we have measured the changes in fluorescence lifetime traces of the fluorescent dye fluorescein covalently bound to the first cytoplasmic loop of bacteriorhodopsin (bR) after light activation of protein function. The fluorescence is excited by a picosecond laser pulse. The retinylidene chromophore of bR is light-activated by a 10 ns laser pulse, which in turn triggers recording of a sequence of fluorescence lifetime traces in the mdTCSPC-module. The fluorescence decay changes upon protein function occur predominantly in the 100 ps time range. The kinetics of these changes shows two transitions between three intermediate states in the second part of the bR photocycle. Correlation with photocycle kinetics allows for the determination of reaction intermediates at the proteins surface which are coupled to changes in the retinal binding pocket. PMID- 17117890 TI - The crystal structure of the L1 intermediate of halorhodopsin at 1.9 angstroms resolution. AB - The mutant T203V of the light driven chloride pump halorhodopsin from Halobacterium salinarum was crystallized and the X-ray structure was solved at 1.6 angstroms resolution. The T203V structure turned out to be nearly identical to the wild type protein with a root mean square deviation of 0.43 angstroms for the carbon alpha atoms of the protein backbone. Two chloride binding (CB) sites were demonstrated by a substitution of chloride with bromide and an analysis of anomalous difference Fourier maps. The CB1 site was found at the same position as in the wild type structure. In addition, a second chloride binding site CB2 was identified around Q105 due to higher resolution in the mutant crystal. As T203V showed a 10 times slower decay of its photocycle intermediate L, this intermediate could be trapped with an occupancy of 60% upon illumination at room temperature and subsequent cooling to 120 degrees K. Fourier transform infrared spectroscopy clearly identified the crystal to be trapped in the L1 intermediate state and the X-ray structure was solved to 1.9 angstroms resolution. In this intermediate, the chloride moved by 0.3 angstroms within binding site CB1 as indicated by peaks in difference Fourier density maps. The chloride in the second binding site CB2 remained unchanged. Thus, intraproteinous chloride translocation from the extracellular to the cytoplasmic part of the protein must occur in reaction steps following the L1 intermediate in the catalytic cycle of halorhodopsin. PMID- 17117891 TI - Differential biodistribution of adenoviral vector in vivo as monitored by bioluminescence imaging and quantitative polymerase chain reaction. AB - A better understanding of the in vivo biodistribution of adenoviral vectors would enable the researcher to anticipate potential side effects due to off-targeted site of transduction, and aid in the strategic design of gene therapy. We combined real-time polymerase chain reaction with in vivo optical imaging to examine viral transduction in liver, lung, spleen, kidney, prostate, and lymph nodes. A replication-deficient serotype 5 adenoviral vector expressing the firefly luciferase gene under the control of a constitutive cytomegalovirus promoter was administered in vivo via different routes. Intravenous and intraperitoneal injections resulted in greatest gene expression and viral DNA in the liver, whereas intraperitoneal injections led to a greater extent of gene delivery to the prostate. Although prostate-directed injection resulted in dominant gene expression in the targeted site, leakage of the vector to other organs was also observed. Vector injection into the lymphatic-rich paw tissue or the subcutaneous tissue of shoulder or chest followed the expected lymphatic drainage pattern, resulting in the accumulation of viral vector in ipsilateral brachial and axillary lymph nodes. Collectively, this study demonstrates that each tissue retains various amounts of adenoviral vector, depending on the route of administration. This knowledge is useful in the strategic design and implementation of adenovirus-mediated gene therapies. PMID- 17117895 TI - Efficacy of oncolytic herpesvirus NV1020 can be enhanced by combination with chemotherapeutics in colon carcinoma cells. AB - NV1020, an oncolytic herpes simplex virus type 1, can destroy colon cancer cells by selectively replicating within these cells, while sparing normal cells. NV1020 is currently under investigation in a clinical phase I/II trial as an agent for the treatment of colon cancer liver metastases, in combination with conventional chemotherapeutic agents such as 5-fluorouracil (5-FU), SN38 (the active metabolite of irinotecan), and oxaliplatin. To study the synergy of NV1020 and chemotherapy, cytotoxicity and viral replication were evaluated in vitro by treating various human and murine colon carcinoma cell lines, using a colorimetric viability assay, a clonogenic assay, and a plaque-forming assay. In vivo experiments, using a subcutaneous syngeneic CT-26 tumor model in BALB/c mice, were performed to determine the efficacy of combination therapy. In vitro studies showed that the efficacy of NV1020 on human colon carcinoma cell lines HT 29, WiDr, and HCT-116 was additively or synergistically enhanced in combination with 5-FU, SN38, or oxaliplatin. The sequence of application was not important and effects were still apparent after a 21-day incubation period. Three intra tumoral treatments with NV1020 (1 x 10(7) plaque-forming units), followed by three subcutaneous treatments with 5-FU (50 mg/kg), resulted in substantially higher inhibition of tumor growth and prolongation of survival compared with monotherapies (NV1020/5-FU vs. NV1020, p = 0.027). On WiDr cells, reduced replication of NV1020, in combination with 5-FU, indicated that additive and synergistic effects of combination therapy must be independent from viral replication. These results suggest that NV1020, in combination with chemotherapy, is a promising therapy for treating patients with metastatic colorectal cancer of the liver. We hypothesize that infection of cells with NV1020 sensitizes the infected cells for the cytotoxic effect of the chemotherapeutics. PMID- 17117896 TI - [Kidney transplantation and organ trade in Peru]. PMID- 17117897 TI - [Practical management of oral antidiabetics in patients with renal disease]. PMID- 17117898 TI - [Arterial blood pressure variations: homocysteine and nitric oxide synthase gene polymorphisms (NOS3)]. AB - BACKGROUND: The nitric oxide synthase (NOS3) G894T gene polymorphism seems as a genetic determinant of total homocysteine (tHcy) concentrations through an effect on folate catabolism. We tested for a significant contribution to blood pressure values for the NOS3 G894T and 4a/b gene polymorphisms and whether those changes could explain the modulating effect on tHcy concentrations. PATIENTS AND METHODS: We analyzed 158 healthy men. The NOS3 gene polymorphisms were determined by polymerase chain reaction (PCR) and restriction fragment analysis (G894T) and by PCR (4a/b). Total homocysteine concentrations were evaluated by the fluorescence polarization immunoassay method. RESULTS: In our population we did not obtain a significant contribution of the G894T to blood pressure variations. However, tHcy mean concentration values differed according G894T genotypes (P = 0.01). Interestingly, we did not obtain a significant modulating effect on tHcy concentrations according to 4a/b genotypes although the 4a/b genotype distribution was statistically associated with blood pressure variations. CONCLUSION: Our results showed a modulating effect of the NOS3 4a/b gene variant on tHcy concentrations that is at least partially provoked by discrete blood pressure increments. Nevertheless, our multivariate analysis did not show a statistical significant role for the NOS3 G894T gene polymorphism on tHcy concentrations. PMID- 17117899 TI - [Hypertension in hemodialysis: prevalence and associated factors in Catalonia. The PRESDIAL study]. AB - OBJECTIVE: To evaluate the prevalence of hypertension (HT) in prevalent hemodialysis (HD) patients in our region, and to analyze the associated clinical and biochemical variables. METHODS: Observational, cross-sectional and multicentric study including a representative sample of prevalent and stable (> 6 months) HD patients from all the HD centers (in and out of Hospitals) in Catalonia, Spain. Clinical and biochemical variables were recorded and predialysis blood pressure (BP) was determined (x3) in each dialysis session during 1 month, as well as the pre/post weight weekly. HT was defined as having at least one of these criteria: a mean (12 determinations) systolic BP > or = 140 mmHg or diastolic BP > or = 90 mmHg or antihypertensive treatment for at least 3 months. RESULTS: The sample comprised 387 patients from 32 of the 40 centers included, 231 of whom where men, with mean age of 63 +/- 14 years. The prevalence of HT in this sample was 67.4%, varying according to the etiology of End-Stage Renal Disease: diabetic 81%, vascular 81%, glomerulonephritis 61%, PKD 52%, unknown and others 64%. The prevalence of additional CV risk factors was 83%. One of each hypertensive 4 patients were treated, of whom 58% had systolic BP > or = 140 or dyastolic > or = 90, in contrast to 28% of untreated patients. The proportion of individuals according to the number of antihypertensive agents was 21% (no agents), 48% (1 agent), 20% (2 agents), 11% (3 agents). Blood pressure was higher among patients receiving higher number of antihypertensive agents. No differences according HT were found in age (64 +/- 13 in hypertensive patients versus 60 +/- 15 in normotensives), time on dialysis (4 +/- 4 vs 4 +/- 4 years), interdialysis weight gain (2.1 +/- 0.8 vs 2.1 +/- 0.8 kg), proportion of weight gain (3.3 +/- 1.4 vs 3.1 +/- 1.4%) or proportion of patients with > 5% weight gain with respect to dry weight (32.5 vs 27.3%). While 84% of hypertensive patients had an additional CV risk factor, this value was 67% in the patients without HT (p < 0.001). CONCLUSION: In HD patients HT has a high prevalence in our region and is poorly controlled. The causes of this poor control may be multiple, and weight gain parameters seem not to be a main factor in these stable patients. Due to the aggregation of risk factors in these patients, strategies in order to improve BP control in HD are mandatory. PMID- 17117900 TI - [Renovascular hypertension in children and adolescents: diagnosis and treatment over 19 years]. AB - SUMMARY Sixty seven hypertensive children age 2-18 with at least one possible clinical sign of renovascular hypertension (RVH) were enrollment into a screening program for diagnose and treatment of RVH over a 19 year period. Patients underwent a variety of biochemical and imaging studies, but in all cases, renal arteriography was used to determine the precise diagnosis and treatment strategy. Of the 67 patients 21 (31.3%) were identified with renal artery stenosis Group 1, 14 (66.6%) unilateral, 5 (23.8%) bilateral and 2 (9.6%) branches. The mean age was 13.9 +/- 3.73 years, with 26.4 +/- 35.2 months of known hypertension, mean systolic blood pressure 191.1 +/- 30.6 mmHg, mean diastolic blood pressure 135.3 +/- 21.2 mmHg and 69% due to fibromuscular dysplasia. Three therapeutic modalities were chosen: percutaneous transluminal angioplasty (PTA), surgery (autotransplant, and nephrectomy), pharmaceutical therapy with antihypertensive drugs and combination of these. The aim of the treatment was blood pressure control, prevention of chronic renal disease and renal function and organ damage preservation. The outcome was categorized as cure, improvement or no change in hypertension. PTA treated eleven patients, 2 combined with surgery (one nephrectomy and 1 autotransplant). Blood pressure was normalized in 9/11 (81.8%) after a mean follow-up of 11.5 years (range 1-18 years). All 6 RVH cases treated by surgery procedure (one after PTA) were cured and 4 cases were managed medically (pharmacological treatment). On december 2004, 19/21 (90%) RVH adolescents blood pressure was normalized with normal serum creatinina, 10 (48%) of these completed cured, 9/21 (43%) improved (normotension with decrease in medication requirements) and 2 (9%) other cases ware lost of follow-up. The 46 non-RVH adolescents (68.7%) were treated with long term antihypertensive medications; all of these have adequate BP control and normal renal function. We conclude that our work-up used in order to make a proper and timely diagnosis and treatment of renovascular hypertension in adolescent was successful in our population. PMID- 17117901 TI - [Prevalence and functional profile of unsuspected radial artery stenosis in native radiocephalic fistula dysfunction. Diagnosis by vascular access flow monitoring using Delta-H method]. AB - OBJECTIVE: The aim of this study was to know the prevalence and functional profile of RA stenosis in RCF dysfunction detected as a result of our surveillance programme. PATIENTS AND METHODS: We prospectively monitored QA of 116 VA (arteriovenous fistula 81% or graft 19%; mean VA duration 28.2 +/- 52.9 months) during hemodiaysis (HD) in 102 ESRD (mean age 63.0 +/- 13.0 yr; sex M: 56.9%, F: 43.1%; mean time on HD 31.4 +/- 44.0 months; 15.5% diabetes) patients (pts) over 4 yr period. QA was measured at least every 4 months by the Delta-H method using the Crit-Line III monitor (overall mean QA 1,193.4 +/- 490.3 ml/min) Forty-three VA (43/116, 37%) met criteria of positive evaluation (absolute QA < 700 ml/min: 48.8%; NQA > 20% from baseline: 51.2%) and were referred for angiography. Most VA explored by angiography showed stenosis > or = 50% (36/40, 90%) that were mainly located in RCF (25/36, 69.4%: RA 11/25, arterialized vein AV 14/25). RESULTS: Eleven cases of RA stenosis (prevalence: 11/36, 30.5%; mean degree: 83.5 +/- 15.8%) were found in 11 RCF (mean VA duration 48.9 +/- 76.7 months) of 11 pts (mean age 67.5 +/- 11.5 yr; mean time on HD 54.0 +/- 75.8 months; 18.2% diabetes). Cause of positive evaluation: absolute QA < 700 ml/min 81.8%; NQA > 20% from baseline 18.2%. Mean QA of RCF just before angiography: 532.9 +/- 99.8 ml/min (range, 418-699 ml/min). Stenosis type: Type I (multiple stenoses) 9.1%, type II (isolated stenosis but critical > 90-95%) 36.4% and type III (isolated stenosis 50-90% with normal haemodynamic status of RCF) 54.5%. Followup: stenosis not reparable 36.4% (4/11), elective intervention by surgery 36.4% (4/11), lost of follow-up before intervention 27.3% (2/11 died, 1/11 transplantation). Mean QA of RCF tended to increase from 547.0 +/- 100.6 ml/min just before surgery to 872.3 +/- 526.5 ml/min just after surgery (n = 4, mean DQA = 325.2 +/- 431.3 ml/min (p = 0.068). Comparative study with 14 AV stenosis (mean degree 76.4 +/- 7.4%) in 11 RCF (mean VA duration 16.4 +/- 22.8 months) of 11 pts (mean age 64.3 +/- 10.5 yr; mean time on HD 17.0 +/- 18.9 months; 50% diabetes): higher prevalence of intervention (85.8%) compared to RA stenosis (p =0.011); without differences in degree of stenosis (p = 0.12) and QA before angiography (p = 0.78) or surgery (p = 1.00); mean QA increased significantly after surgery (n = 6 AV, 549.8 +/- 86.4 vs 1,033.0 +/- 216.6 ml/min) (p = 0.028). CONCLUSIONS: 1) One third of cases of VA dysfunction were related to feeding artery stenosis. 2) No differences in functional profile were found between RA and AV stenosis before angiography and surgery. 3) The functional results of elective surgery in RA stenosis were worse compared to AV stenosis. PMID- 17117902 TI - [Vascular access for haemodyalisis. Comparative analysis of the mechanical behaviour of native vessels and prosthesis]. AB - INTRODUCTION: The prosthesis nowadays used in the vascular access for haemodialysis have low patency rates, mainly due to the luminal obstruction, determined by the intimal hyperplasia. Several factors have been related to de development of intimal hyperplasia and graft failure. Among them are the differences in the biomechanical properties between the prosthesis and the native vessels. In the searching for vascular prosthesis that overcomes the limitations of the currently used, the cryopreserved vessels (cryografts) appear as an alternative of growing interest. However, it is unknown if the mechanical differences or mismatch between prosthesis and native vessels are lesser when using cryografts. OBJECTIVE: To characterize and compare the biomechanical behaviour of native vessels used in vascular access and cryografts. Additionally, segments of expanded polytetrafluoroethylene (ePTFE) were also evaluated, so as to evaluate the potential biomechanical advantages of the cryografts respect to synthetic prosthesis used in vascular access. METHODS: Segments from human humeral (n = 12), carotid (n = 12) and femoral (n = 12) arteries, and saphenous vein (n = 12), were obtained from 6 multiorgan donors. The humeral arteries were studied in fresh state. The other segments were divided into two groups, and 6 segments from each vessel were studied in fresh state, while the remaining 6 segments were evaluated after 30 days of criopreservation. For the mechanical evaluation the vascular segments and 6 segments of ePTFE were mounted in a circulation mock and submitted to haemodynamic conditions similar to those of the in vivo. Instantaneous pressure (Konigsberg) and diameter (Sonomicrometry) were measured and used to calculate the viscous and elastic indexes, the compliance, distensibility and characteristic impedance. For each mechanical parameter studied, the mismatch between the prosthesis and the native vessel was evaluated. RESULTS: The ePTFE was the prosthesis with the higher mechanical mismatch (p < 0.05). The venous and arterial cryografts showed the least mismatch with native veins and arteries, respectively. The prosthesis with the least mechanical mismatch was different, depending on the native vessel evaluated, and for a native vessel, on the parameter considered. CONCLUSION: The mechanical mismatch between the native vessel and the vascular prosthesis used in a vascular access could be reduced using cryografts. PMID- 17117903 TI - [Comparative study of PTFE grafts in forearm vs cuffed permanent catheters]. AB - INTRODUCTION: As is universally accepted the best form of permanent vascular access for haemodialysis is the native arteriovenous fistula. A second and third options are the politetrafluoroethylene (PTFE) AV grafts and the cuffed, tunneled, internal catheters. The overall performance and complications of catheters is clearly inferior to AV fistula. There are not many studies that compare permanent catheters to grafts in terms of functionality, survival and complications. METHODS: We analyzed 81 vascular accesses carried out from october 99 to december 03 in 59 patients and during a follow-up period of 35 months. Two groups were considered. Group 1, catheters (n 42) and group 2, grafts (n 39). Clinical aspects, comorbidity index (Wright and Kanh), dialysis dose and complications and survival of the access were registered. RESULTS: Both groups were similar in age, sex, time on haemodialysis, number of previous accesses and hospitalization days. Cardiovascular morbidity and comorbidity index were significantly higher in patients with catheter. While blood flow during dialysis was higher in grafts both groups showed no significant differences in parameters of efficacy of dialysis (Kt/V, TAC BUN and PCRn). Serum albumin was lower in patients with catheter. The number of accesses that failed was higher in the graft group being thrombosis the main complication followed by infection. Kaplan Meier curves showed better accumulated survival of permanent catheters versus grafts (61,4% vs 9,8% at 35 months). The most frequent complication of catheter was infection while in the case of grafts it was thrombosis followed by infection. CONCLUSIONS: Although they were placed in patients with higher comorbidity, cuffed, tunneled catheters showed less number of complications and better survival than PTFE grafts in our patients in haemodialysis. The main cause of failure of both vascular access was thrombosis followed by infection. The dose of dialysis obtained was no different in both groups. Cuffed, tunneled permanent catheters are a very interesting option in a number of patients in haemodialysis and they can be an option to consider in those patients with vascular difficulties and higher comorbidities. PMID- 17117904 TI - [Functional dependency evaluation of hemodialysis patients: a multicentric study]. AB - BACKGROUND: There has been a change in the hemodialysis population characteristics over the last years with a progressive increase in patient,s age and associated comorbility and mortality. This older hemodialysis population are more functionally and medically dependent increasing the time taken to perform nursing work. The objective of this study was to evaluate the degree of functional dependency on hemodialysis patients and the need of care by nursing workload. METHODS: A transversal descriptive study was done during 1 month (april 2005) on 586 patients from 10 HD Units in Spain. No exclusion criteria were used. The Delta Test, who was used to evaluate the patients dependency needs, is a workload measure instrument base on three subscales: dependency, Physical Deficiency and mental deficiency. The indicators are measure on a scale of one to three with each level representing an increasing demand on nursing time. The results obtained from the Delta Test were analyzed taking into account the following variables: age, average time in dialysis, Charlson comorbidity Index, geographic location and HD unit. RESULTS: 46% of the patients show some degree of dependency, of these 12.8% were moderated and 8.1% severe; the subscale analysis showed that 19.6% and 6.7% had, respectively, a moderate to severe physical and mental health deficiency. The dependency degree varied significantly between HD Units and geographic location with a range of 0% to 59.8%. The degree of dependency were statistically associated with age and CCI. The higher CCI (r: 0,21; p < 0.001) and age (r: 0,26; p < 0.001) the higher was the Delta Test Score for dependency level. Patients times of initiation on dialysis were not associated with an increase in the degree of dependency. The aspects evaluated by the Delta Test that showed a higher score were those related to patients mobility. The patients assistance requirements during the HD session are basically related to a lack of mobility due to musculoskeletal disease and to a lesser extent to behavior alterations. CONCLUSIONS: HD units are attending patients with an important degree of dependency which impose and added workload to the healthcare personnel. In occasions, it is very difficult to attend and give proper care with the current legally established nurse to patient ratio which. This suggests the need to implement a better staffing policy. The Delta Test provides an objective, adaptable and standardized instrument for measuring degree of dependency of HD patients. PMID- 17117905 TI - [Acute rejection of kidney transplant is associated with induction of replicative senescence in CD8+ T lymphocytes]. AB - Acute renal rejection repeatedly activates immunocompromised CD8 + T cells. Maintained activation of CD8 + T cells can induce a process of replicative senescence. In the present study, we will evaluate in CD8 lymphocytes from patients undergoing acute renal rejection characteristics of replicative senescence such as: a) low expression of CD28 molecule; b) telomere shortening and c) increase production of proinflammatory cytokines. The study was carried out in CD8 + T cells from 14 patients transplanted without clinical evidences of acute renal rejection, 14 patients kidney transplanted with clinical and anatomopathological evidences of acute renal rejection, 8 healthy controls. The results shown that in peripheral blood and renal biopsy of patients with acute renal rejection there is a significant increment of the population of T cells CD28-CD8+, with short telomere length, as compared with healthy controls and patients without acute renal rejection. The presence of senescent cells was associated with high levels of IL-10 and IFN-Y in plasma and urine. In conclusion our study suggest that the CD8 + T cells of patients with acute renal rejection suffer a process of replicative senescence. PMID- 17117906 TI - [Concordance of Cockcroft Gault formula and MDRD formula to estimate glomerular filtration rate in patients with diabetes type 2]. PMID- 17117907 TI - [Perforation of the acalculous gallbladder in a renal transplant recipient with CMV infection]. AB - We report the case of a renal transplant recipient who developed acute acalculous cholecyscitis resulting in gallbladder perforation. At admission CMV antigenemia was negative. Emergency laparatomy was performed and showed the gallbladder to be infarted with a perforation. The abdominal cavity contained two litres of sterile bilious fluid. The pathological report showed frequent endothelial cells contained intranuclear and intracitoplasmatic inclusion (fig. 1). Treatment with Ganciclovir iv was started after diagnosis, but a computerized tomography scan demonstrated severe acute pancreatitis (grade E. Baltazar). The patient developed multiorgan failure and died on 19th day after surgery. Necropsy showed cytomegalic inclusions in pancreas (fig. 2), gastrointerstinal tract, lung and graft. A necroticing pneumonia with Mycotic spores and hiphae was seen. Aspergillus was also observed in myocardium (fig. 3). PMID- 17117908 TI - [A case of ureteritis in Henoch-Shonlein purpura]. AB - Henoch-Schonlein purpura (HSP) is a multisystemic disorder mainly affecting the skin, joints, gastrointestinal tract, and kidneys but sometime rare complications that have been reported include urologic manifestations. We report a case of a 63 year old man was admitted to the hospital because a neurologic abnormalities, gastrointestinal blood loss and acute renal failure. One year before HSP was dignosed by percutaneous renal biopsy. A few days later admission the temperature was 38 masculineC and urine cultures yielded Echerichia Coli. An angio-magnetic resonance of the abdomen showed a marked dilatation of the distal left ureter with a level inside. Therapy with corticosteroids improved a clinical features and the usual renal function was recovered. PMID- 17117909 TI - [Acute renal failure and proximal renal tubular dysfuntion in a patient with acquired immunodeficiency syndrome treated with tenofovir]. AB - Tenofovir, a new nucleotide reverse transcriptase inhibitor that has good antiviral activity against drug-resistant strains of HIV, is structurally similar to cidofovir and adefovir and seems to be less nephrotoxic. Nephrotoxicity of cidofovir and adefovir is well established and they have been associated with increase for acute renal insufficiency due to tubular toxicity, possibly induced via mitochondrial deplection. Tenofovir has little mithocondrial toxicity in in vitro assays and early clinical studies. However some cases of renal tubular dysfuntion and renal failure related to tenofovir treatment have been published recently. Increased plasma concentrations of didanosine were observed after the adition of tenofovir and protease inhibitors can interact with the renal transport of organic anions leading to proximal tubular intracellular accumulation of tenofovir, yield Fanconi syndrome-type tubulopathy. We present a case in wich acute renal failure and proximal tubular dysfunction developed after therapy with tenofovir in a patiente with HIV who had suffered from complications of didanosine treatment. Although nephrotoxicity certainly occurs much less frequently with tenofovir that it does with other nuclotide analogues, use of tenofovir by patients with underlying renal disfuntion, for longer durations and/or associated with didanosine or lopinavir-ritonavir, might be associated with renal toxicity. Patients receiving tenofovir must be monitored for sings of tubulopathy with simple tests such us glycosuria, phosphaturia, proteinuria, phosphoremia and renal function, as well as assessment for signs of mithocondrial toxicity when a nucleoside analogue is being administered, and therapy should be stopped to avoid the risk of definitive renal failure. PMID- 17117910 TI - [Diagnosis of sarcoidosis from the study of a case with acute renal failure]. PMID- 17117911 TI - [Accelerated arterial hypertension due to intradermal anovulatory drugs]. PMID- 17117912 TI - [The biopsychosocial integrating view as a strategy for the patient with chronic renal failure (CRF): a contemporary requisite]. PMID- 17117913 TI - [Microalbuminuaria. Renal and cardiovascular risk factor]. PMID- 17117914 TI - [Cytomegalovirus infection-associated thrombotic microangiopathy as the first clinical manifestation of HIV primary infection]. PMID- 17117915 TI - [Prototheca wickerhamii peritonitis in peritoneal dialysis patients]. PMID- 17117916 TI - [Prevalence of occult chronic renal disease in hypertensive patients]. PMID- 17117917 TI - [Management of nephrotic syndrome with tacrolimus (FK 506)]. PMID- 17117918 TI - [Maxillary brown tumor in secondary hyperparathyroidism]. PMID- 17117919 TI - [Penis necrosis in a diabetic patient on hemodialysis]. PMID- 17117920 TI - Proteasome proteolytic activity in skeletal muscle is increased in patients with sepsis. AB - Patients with sepsis in the ICU (intensive care unit) are characterized by skeletal muscle wasting. This leads to muscle dysfunction that also influences the respiratory capacity, resulting in prolonged mechanical ventilation. Catabolic conditions are associated with a general activation of the ubiquitin proteasome pathway in skeletal muscle. The aim of the present study was to measure the proteasome proteolytic activity in both respiratory and leg muscles from ICU patients with sepsis and, in addition, to assess the variation of proteasome activity between individuals and between duplicate leg muscle biopsy specimens. When compared with a control group (n=10), patients with sepsis (n=10) had a 30% (P<0.05) and 45% (P<0.05) higher proteasome activity in the respiratory and leg muscles respectively. In a second experiment, ICU patients with sepsis (n=17) had a 55% (P<0.01) higher proteasome activity in the leg muscle compared with a control group (n=10). The inter-individual scatter of proteasome activity was larger between the patients with sepsis than the controls. We also observed a substantial intra-individual difference in activity between duplicate biopsies in several of the subjects. In conclusion, the proteolytic activity of the proteasome was higher in skeletal muscle from patients with sepsis and multiple organ failure compared with healthy controls. It was shown for the first time that respiratory and leg muscles were affected similarly. Furthermore, the variation in proteasome activity between individuals was more pronounced in the ICU patients for both muscle types, whereas the intra-individual variation between biopsies was similar for ICU patients and controls. PMID- 17117921 TI - NAADP induces pH changes in the lumen of acidic Ca2+ stores. AB - NAADP (nicotinic acid-adenine dinucleotide phosphate)-induced Ca2+ release has been proposed to occur selectively from acidic stores in several cell types, including sea urchin eggs. Using fluorescence measurements, we have investigated whether NAADP-induced Ca2+ release alters the pH(L) (luminal pH) within these acidic stores in egg homogenates and observed their prompt, concentration dependent alkalinization by NAADP (but not beta-NAD+ or NADP). Like Ca2+ release, the pH(L) change was desensitized by low concentrations of NAADP suggesting it was secondary to NAADP receptor activation. Moreover, this was a direct effect of NAADP upon the acidic stores and not secondary to increases in cytosolic Ca2+ as it was not mimicked by IP3 (inositol 1,4,5-trisphosphate), cADPR (cyclic adenine diphosphoribose), ionomycin, thapsigargin or by direct addition of Ca2+, and was not blocked by EGTA. The results of the present study further support acidic stores as targets for NAADP and for the first time reveal an adjunct role for NAADP in regulating the pH(L) of intracellular organelles. PMID- 17117922 TI - Identification of novel phosphorylation sites in MSK1 by precursor ion scanning MS. AB - MSK1 (mitogen- and stress-activated kinase 1) is a dual kinase domain protein that acts downstream of the ERK1/2 (extracellular-signal-regulated kinase 1/2) and p38 MAPK (mitogen-activated protein kinase) signalling pathways in cells. MSK1, and its related isoform MSK2, phosphorylate the transcription factors CREB (cAMP-response-element-binding protein) and ATF1 (activating transcription factor 1), and the chromatin proteins histone H3 and HMGN1 (high-mobility-group nucleosomal-binding protein 1) in response to either mitogenic stimulation or cellular stress. MSK1 activity is tightly regulated in cells, and activation requires the phosphorylation of MSK1 by either ERK1/2 or p38a. This results in activation of the C-terminal kinase domain, which then phosphorylates further sites in MSK1, leading to the activation of the N-terminal kinase domain and phosphorylation of substrates. Here, we use precursor ion scanning MS to identify five previously unknown sites in MSK1: Thr630, Ser647, Ser657, Ser695 and Thr700. One of these sites, Thr700, was found to be a third site in MSK1 phosphorylated by the upstream kinases ERK1/2 and p38a. Mutation of Thr700 resulted in an increased basal activity of MSK1, but this could be further increased by stimulation with PMA or UV-C radiation. Surprisingly, however, mutation of Thr700 resulted in a dramatic loss of Thr581 phosphorylation, a site essential for activity. Mutation of Thr700 and Thr581 to an alanine residue resulted in an inactive kinase, while mutation of both sites to an aspartic acid residue resulted in a kinase with a significant basal activity that could not be further stimulated. Together these results are consistent with a mechanism by which Thr700 phosphorylation relieves the inhibition of MSK1 by a C-terminal autoinhibitory helix and helps induce a conformational shift that protects Thr581 from dephosphorylation. PMID- 17117923 TI - Some characteristics and purification of anti-(human ovarian carcinoma)xanti (human CD3) single-chain bispecific antibody. AB - To obtain high-quality AhOC x AhCD3 [anti-(human ovarian carcinoma)xanti-(human CD3)] with high biological activity economically and easily, some characteristics and purification of AhOC x AhCD3, a single chain bispecific antibody, were investigated. During the present study, some important properties of AhOC x AhCD3, such as molecular mass, pI, solubility, stability, hydrophobic ability, molecular status and bioactivity were primarily studied. The molecular mass of AhOC x AhCD3 was determined to be approx. 58.0 kDa by SDS/PAGE (theoretical molecular mass: 56972.5 Da, calculated on the basis of the primary structure) and the pI was approx. 7.5+/-0.4 by IEF (isoelectric focusing; theoretical pI: 8.5, predicted by the ProtParam software tool) respectively. Experiments showed that the solubility of AhOC x AhCD3 increased with the pH increase and that the AhOC x AhCD3 was easily degraded into several fragments during the storage time of samples and the mixtures of both fragment B and fragment C retained normal bioactivity. Particularly, the soluble aggregate/polymer status of AhOC x AhCD3 with bioactivity was verified by non-reducing SDS/PAGE. After optimization of purification process, AhOC x AhCD3 with electrophoretic homogeneity was successfully obtained and the yield was approx. 20%. Some important properties of AhOC x AhCD3 were first observed and a procedure for soluble AhOC x AhCD3 purification in scale-up was first established. Some characteristics of AhOC x AhCD3, especially the stability of AhOC x AhCD3, are critical for the development of this protein pharmaceutical and useful for reference to other proteins. PMID- 17117924 TI - Endosomal receptor kinetics determine the stability of intracellular growth factor signalling complexes. AB - There is an emerging paradigm that growth factor signalling continues in the endosome and that cell response to a growth factor is defined by the integration of cell surface and endosomal events. As activated receptors in the endosome are exposed to a different set of binding partners, they probably elicit differential signals compared with when they are at the cell surface. As such, complete appreciation of growth factor signalling requires understanding of growth factor receptor binding and trafficking kinetics both at the cell surface and in endosomes. Growth factor binding to surface receptors is well characterized, and endosomal binding is assumed to follow surface kinetics if one accounts for changes in pH. Yet, specific binding kinetics within the endosome has not been examined in detail. To parse the factors governing the binding state of endosomal receptors we analysed a whole-cell mathematical model of epidermal growth factor receptor trafficking and binding. We discovered that the stability of growth factor-receptor complexes within endosomes is governed by three primary independent factors: the endosomal dissociation constant, total endosomal volume and the number of endosomal receptors. These factors were combined into a single dimensionless parameter that determines the endosomal binding state of the growth factor-receptor complex and can distinguish different growth factors from each other and different cell states. Our findings indicate that growth factor binding within endosomal compartments cannot be appreciated solely on the basis of the pH dependence of the dissociation constant and that the concentration of receptors in the endosomal compartment must also be considered. PMID- 17117925 TI - Solution structure of family 21 carbohydrate-binding module from Rhizopus oryzae glucoamylase. AB - CBMs (carbohydrate-binding modules) function independently to assist carbohydrate active enzymes. Family 21 CBMs contain approx. 100 amino acid residues, and some members have starchbinding functions or glycogen-binding activities. We report here the first structure of a family 21 CBM from the SBD (starch-binding domain) of Rhizopus oryzae glucoamylase (RoCBM21) determined by NMR spectroscopy. This CBM has a beta-sandwich fold with an immunoglobulin-like structure. Ligand binding properties of RoCBM21 were analysed by chemical-shift perturbations and automated docking. Structural comparisons with previously reported SBDs revealed two types of topologies, namely type I and type II, with CBM20, CBM25, CBM26 and CBM41 showing type I topology, with CBM21 and CBM34 showing type II topology. According to the chemical-shift perturbations, RoCBM21 contains two ligand binding sites. Residues in site II are similar to those found in the family 20 CBM from Aspergillus niger glucoamylase (AnCBM20). Site I, however, is embedded in a region with unique sequence motifs only found in some members of CBM21s. Additionally, docking of beta-cyclodextrin and malto-oligosaccharides highlights that side chains of Y83 and W47 (one-letter amino acid code) form the central part of the conserved binding platform in the SBD. The structure of RoCBM21 provides the first direct evidence of the structural features and the basis for protein-carbohydrate recognition from an SBD of CBM21. PMID- 17117926 TI - Post-translational palmitoylation and glycosylation of Wnt-5a are necessary for its signalling. AB - Wnt-5a is a representative ligand that activates a beta-catenin-independent pathway in Wnt signalling. In the present paper, the roles of the post translational modifications in the actions of Wnt-5a were investigated. We found that Wnt-5a is modified with palmitate at Cys104 and glycans at Asn114, Asn120, Asn311 and Asn325. The palmitoylation was not essential for the secretion of Wnt 5a, but was necessary for its ability to suppress Wnt-3a-dependent T-cell factor transcriptional activity and to stimulate cell migration. Wnt-5a activated focal adhesion kinase and this activation also required palmitoylation. Wild-type Wnt 5a induced the internalization of Fz (Frizzled) 5, but a Wnt-5a mutant that lacks the palmitoylation site did not. Furthermore, the binding of Wnt-5a to the extracellular domain of Fz5 required palmitoylation of Wnt-5a. These results indicate that palmitoylation of Wnt-5a is important for the triggering of signalling at the cell surface level and, therefore, that the lipid-unmodified form of Wnt-5a cannot activate intracellular signal cascades. In contrast, glycosylation was necessary for the secretion of Wnt-5a, but not essential for the actions of Wnt-5a. Thus the post-translational palmitoylation and glycosylation of Wnt-5a are important for the actions and secretion of Wnt-5a. PMID- 17117927 TI - 1.8 A bright-state structure of the reversibly switchable fluorescent protein Dronpa guides the generation of fast switching variants. AB - RSFPs (reversibly switchable fluorescent proteins) may be repeatedly converted between a fluorescent and a non-fluorescent state by irradiation and have attracted widespread interest for many new applications. The RSFP Dronpa may be switched with blue light from a fluorescent state into a non-fluorescent state, and back again with UV light. To obtain insight into the underlying molecular mechanism of this switching, we have determined the crystal structure of the fluorescent equilibrium state of Dronpa. Its bicyclic chromophore is formed spontaneously from the Cys62-Tyr63-Gly64 tripeptide. In the fluorescent state, it adopts a slightly non-coplanar cis conformation within the interior of a typical GFP (green fluorescent protein) b-can fold. Dronpa shares some structural features with asFP595, another RSFP whose chromophore has previously been demonstrated to undergo a cis-trans isomerization upon photoswitching. Based on the structural comparison with asFP595, we have generated new Dronpa variants with an up to more than 1000-fold accelerated switching behaviour. The mutations which were introduced at position Val157 or Met159 apparently reduce the steric hindrance for a cis-trans isomerization of the chromophore, thus lowering the energy barrier for the blue light-driven on-to-off transition. The findings reported in the present study support the view that a cis-trans isomerization is one of the key events common to the switching mechanism in RSFPs. PMID- 17117928 TI - Current direction the nursing profession. PMID- 17117929 TI - Using NANDA, NIC, and NOC (NNN) language for clinical reasoning with the Outcome Present State-Test (OPT) model. AB - PURPOSE: To analyze the degree to which standardized nursing language was used by baccalaureate nursing students completing Outcome-Present State-Test (OPT) model worksheets in a clinical practicum. METHODS. A scoring instrument was developed and 100 worksheets were retrospectively analyzed. FINDINGS: NANDA nursing diagnoses were correctly stated in 92% of the OPT models. Nursing Outcomes Classification (NOC) outcomes were explicitly stated in 22%, and implied in 72%. Interventions matched appropriate Nursing Interventions Classification (NIC) activities in 61%. CONCLUSIONS: NANDA, NIC, and NOC (NNN) language was used inconsistently by students in this sample. IMPLICATIONS FOR PRACTICE: If NNN language is to advance nursing knowledge, its promotion, representation in curriculum development, and active use is necessary. Educational research is needed on the facilitators and barriers to NNN language use. PMID- 17117930 TI - Nursing diagnoses in a Brazilian intensive care unit. AB - PURPOSE: To identify the nursing diagnoses and their most frequent related factors or risk factors in patients admitted to an intensive care unit (ICU). METHOD: Descriptive cross-sectional study with information from 991 admissions to an ICU during a 6-month period. FINDINGS: Sixteen nursing diagnoses resulting from hospitalization were most frequently identified; six had percentages greater than 40% with 29 related/risk factors. The resulting averages were 6.9 diagnoses per hospitalization and 1.2 related/risk factors per nursing diagnoses. CONCLUSIONS: The nursing diagnoses identified seemed to be common to the clinical practice of nursing and their fundamental related/risk factors to precise clinical judgment, thus providing a basis for interventions for a desired outcome. PRACTICE IMPLICATIONS: The findings have contributed to the development of the standardized nursing language usage in Brazilian nursing practices. PMID- 17117935 TI - Novel sites of aldose reductase immunolocalization in normal and streptozotocin diabetic rats. AB - Glucose metabolism by aldose reductase (AR) is implicated in the pathogenesis of many diabetic complications, including neuropathy. We have re-evaluated the distribution of AR in the sciatic nerve and dorsal root ganglion (DRG) of normal rats, expanded these observations to describe the location of AR in the spinal cord and footpad skin, and investigated whether diabetes alters the distribution of AR. In sciatic nerve, AR was restricted to cytoplasm of myelinated Schwann cells and endothelial cells of epineurial, but not endoneurial, blood vessels. AR immunoreactivity (IR) was present in satellite cells in the DRG. In skin, AR-IR was detected in vascular endothelial cells, Schwann cells of myelinated fibers, and axons of perivascular sympathetic nerves. AR was localized selectively to oligodendrocytes of the white matter of spinal cord. The distribution of AR-IR in sciatic nerve, DRG, skin, and spinal cord was not altered by up to 12 weeks of streptozotocin-induced diabetes. Identification of perineuronal satellite cells, oligodendrocytes, and perivascular sympathetic nerves as AR-expressing cells reveals them as cellular sites with the potential to contribute to diabetic neuropathy. PMID- 17117936 TI - The effect of antibodies to TGF-beta1 and TGF-beta2 at a site of sciatic nerve repair. AB - Scar formation at a site of nerve injury can cause a mechanical barrier to axonal regeneration and lead to the development of multiple axonal sprouts to form a neuroma. We have investigated the hypothesis that the application of a scar preventing agent to a nerve repair site would enhance regeneration of the nerve and reduce neuroma formation. The left sciatic nerve was exposed under general anaesthesia in 18 adult Sprague-Dawley rats. In 12 animals, the nerve was sectioned and immediately re-approximated using four epineurial sutures, and in 6 of these animals neutralising antibodies to transforming growth factor (TGF) beta1 and TGF-beta2 were injected into and around the repair site. The six other animals acted as controls. After 7 weeks, the outcome was assessed by recording compound action potential (CAP) ratios, measuring collagen levels using picrosirius red staining, and counting the number of myelinated axons proximal and distal to the repair. After repair alone, the mean percentage of area of staining (PAS) for collagen within the nerve had significantly increased. However, after repair with the administration of antibodies, the PAS was not significantly different from that in the sham controls. After administration of antibodies, the CAP ratios were significantly smaller than in controls but not after repair alone. In both nerve injury groups, the myelinated fibre counts were significantly increased distal to the injury site, but there was no difference between these two groups. We conclude that administration of antibodies to TGF beta1 and TGF-beta2 reduced scar formation at the repair site but did not enhance regeneration of the nerve or reduce the development of multiple axonal sprouts. PMID- 17117937 TI - Altered pain behavior and regeneration after nerve injury in TNF receptor deficient mice. AB - The pro-inflammatory cytokine tumor necrosis factor (TNF)-alpha is an important mediator in hyperalgesia, nerve injury, and regeneration. Here, we used mice deficient of TNF receptor (TNFR) 1 or 2 to investigate the role of TNF signaling via receptor in each pain behavior and nerve de- and regeneration after chronic constriction injury (CCI) of the sciatic nerve. We found an absence of thermal hyperalgesia in mice deficient of TNFR1 and a reduction in mechanical and cold allodynia in mice deficient of TNFR1 or TNFR2 compared with wild-type mice. Nerve conduction studies and nerve pathology did not reveal major differences between genotypes in the temporal course of de- and regeneration of the nerve. We propose that the functional effects of the TNFRs on pain symptoms are independent of effects on nerve regeneration. Furthermore, the differential action of TNF via each of its receptors should be taken into account when considering clinical trials with TNF inhibitors for pain. PMID- 17117938 TI - Acute heroin-related neuropathy. AB - Heroin-related peripheral nervous injury has scarcely been reported, mostly as compressive neuropathy. Rarely, other types of peripheral nervous system (PNS) injury have been recognized, such as plexopathy, polyradiculopathy, mononeuropathy, and rhabdomyolysis. These complications are usually not related to local trauma, but the nature of nerve injury remains unknown. Immunologic mechanisms have been proposed, although generally there is no laboratory evidence of inflammation and usually there is no improvement following steroid therapy. We describe six patients who developed acute PNS injury following intravenous or intranasal heroin self-administration with no evidence of compression injury or inflammation. Four patients had plexopathy (two lumbosacral and two brachial), and two had symmetric distal axonal sensorimotor neuropathy affecting the lower extremities. Of the six patients, five had concomitant rhabdomyolysis (creatine kinase, CK: 5,000-100,000 U/l) and one patient with brachial plexopathy had normal CK levels. The neurological deficit was noticed 3-36 h after heroin administration. Electromyography in five patients was consistent with sensorimotor axonal loss either confined to the affected plexus or with a diffuse distribution in the legs in the two patients with neuropathy. We propose that a toxic mechanism may be responsible for non-compression cases of acute neuropathy following heroin abuse. PMID- 17117939 TI - Nerve root degeneration and regeneration by intrathecal phenol in rats: a morphologic approach. AB - Intrathecal injection of phenol (ITP) has been used to control intractable pain and spasticity. Direct caustic nerve damage has been postulated as the mechanism of analgesia. Sensation is commonly recovered, suggesting that a spontaneous regeneration process takes place. There is, however, a lack of mechanistic information on ITP therapy. To define morphologically the neurolysis and regeneration phenomena produced by ITP, anesthetized rats were subjected to laminectomy at L5; 5 microl of 22% phenol in saline solution or vehicle (control) was injected. Light and electron microscopy studies of nerve roots were performed at 2, 14, and 60 days after injection. Rats given ITP showed at the early stage a variable amount of roots with signs of infarction characterized by loss of axon myelin units and thrombosis of intra-root vessels. At 14 days, abundance of macrophages removing debris, open vessels, and nerve sprouts was identified in damaged roots. At this time, non-myelinating glial fibrillary acidic protein positive Schwann cells were observed in both damaged and apparently undamaged roots. At 60 days, abundance of 2',3'-cyclic nucleotide 3'-phosphodiesterase positive Schwann cells myelinating newly formed axons was observed in damaged roots. Control rats did not show signs of neural or vascular pathology. Attempting to prevent thrombosis, another group of rats received heparin before ITP; these anti-coagulated rats developed radicular thrombosis, neurolysis, and hemorrhage. In conclusion, neurolysis produced by ITP is associated with acute ischemia (not prevented by heparin) and is followed by vascular, nerve, and myelin regeneration. Our results help understand the lack of efficacy of and some complications by ITP clinical therapy. PMID- 17117940 TI - Expression of the antioxidant enzyme peroxiredoxin 5 in the human peripheral nervous system. AB - Peroxiredoxins (PRDXs) belong to a family of peroxidases that are highly expressed in mammalian tissues. Comparatively, little is known about their expression in the nervous system. In this study, we examined in human sural nerve, the expression of PRDX5, the most recently defined peroxiredoxin family member. Immunohistochemical staining of paraffin sections showed that PRDX5 expression was mainly localised to the axon and, to a lesser extent, to the Schwann cells. Immunogold labelling electron microscopy further revealed that PRDX5 is widely distributed in multiple cellular compartments, most importantly in mitochondria but also in axoplasm, Schwann cell cytosol, nucleus, and myelin. A progressive decline in PRDX5 expression with advancing age was identified in human nerves at different ages. The wide distribution of PRDX5 in cellular and sub-cellular structures indicates that it may play important roles in a wide range of tissue components in normal peripheral nerves. PMID- 17117941 TI - Dose of intravenous immunoglobulins in chronic inflammatory demyelinating polyneuropathy. AB - The usual initiating dose of intravenous immunoglobulins (IVIg) in the treatment of chronic inflammatory demyelinating polyneuropathy (CIDP) is 2 g/kg/course. Although not evidence based, subsequent reductions are advised to the lowest possible level for maintenance. In practice, the achievable levels of such reductions and their impact on treatment frequency have not been studied. Factors determining maximal dosage reduction are unknown. We retrospectively reviewed data concerning IVIg therapy for 15 patients with CIDP, from their medical records between 1997 and 2005. Lowest effective dose and treatment frequency were determined. The following correlations were ascertained: dose to frequency, dose to weight, dose to disease duration, amplitude of dose reduction to disease duration, and dose to pre-therapeutic disease severity. Dose reductions were possible in all (mean: 63.3%, range: 42.4-88%). The lowest effective dose of IVIg per course and treatment frequency were both very variable (18-108 g and 2-17 weeks, respectively). Lowest dose per course did not correlate to weight, frequency of administration, disease duration, or pre-therapeutic degree of disability. Amplitude of dose reduction achieved was independent of disease duration. Treatment frequency could not be lowered in any patient. Our findings show that IVIg target doses should be titrated individually but suggest that infusion frequencies are fixed in each case in relapsing CIDP. Importantly, lower dose treatment is not associated with shorter intervals between courses, and lowest effective dose is independent of weight and disease duration. Initial level of disability does not appear to influence dose required. These results suggest considerably lower, standardized, initiating, and maintenance doses might be effective and highlight the need for prospective dose comparative trials. PMID- 17117942 TI - Artemin has potent neurotrophic actions on injured C-fibres. AB - In this study, we have investigated the effects of artemin (ARTN), one of the glial cell line-derived neurotrophic factor (GDNF) family of neurotrophic factors, on C-fibres following nerve injury in the adult rat. GDNF family receptor alpha (GFRalpha) 3, the ligand binding domain of the ARTN receptor, is expressed in 34% of dorsal root ganglion (DRG) cells, predominantly in the peptidergic population of C-fibres and in a proportion of the isolectin B4 (IB4) binding population. Interestingly, only 30% of GFRalpha3-expressing DRG cells co expressed RET (the signal transducing domain). In agreement with previous studies, treatment with ARTN prevented many of the nerve injury-induced changes in the histochemistry of both the peptidergic and the IB4-binding populations of small, but not large, diameter DRG cells. In addition, ARTN treatment maintained C-fibre conduction velocity, and C-fibre evoked substance P release within the dorsal horn following nerve injury. ARTN was also protective following capsaicin treatment, which produces selective C-fibre injury. Given the potent neurotrophic actions of ARTN on C-fibres, it may therefore provide potential for the treatment of nerve injury, particularly in the maintenance of small fibre function. PMID- 17117943 TI - Isolated amyloidosis presenting with lumbosacral radiculoplexopathy: description of two cases and pathogenic review. AB - In this study, we present two cases of infiltrative, localized amyloidosis involving lumbosacral root and plexus, e.g., isolated amyloidomas. Rare and poorly understood amyloidomas may occur in both neurologic and non-neurologic tissues. The described cases emphasize potential for localized peripheral amyloidomas: (1) potential for associated lambda light chain lymphoplasmacytic lymphoma association; (2) e isolated amyloidosis without evidence for systemic plasma cell dyscrasia; (3) features suggestive of potential pathogenesis; and (4) discussion of treatment options including immunotherapy and resection. The limited literature and experience among other cases is described. PMID- 17117944 TI - Steroid-responsive demyelinating motor neuropathy with conduction block associated with hepatitis C infection and treatment with interferon alpha. PMID- 17117945 TI - Cross-mapping between three terminologies with the international standard nursing reference terminology model. AB - PURPOSE: The International Standards Organization's (ISO) International Standard IS 18104 should assist the nursing profession to integrate their terminologies into computer systems and healthcare reference terminologies. The purpose of this study is to cross-map between different terminologies; that is, to determine if concepts in one terminology are similar to concepts in another terminology. METHODS: The ISO standard was used to test the degree to which three terminologies could be cross-mapped to each other. Concepts and terms were selected, their equivalence determined by experts, and the specific concepts were dissected or broken down to their constituent parts. RESULTS: Based on experts' selections from the three classifications, equivalent concepts were identified. Those concepts deemed equivalent were dissected, thus revealing whether the components of the nursing diagnostic concepts such as focus, judgment, and other attributes of the ISO standard matched. Based on the dissection of each diagnosis, the decision was made whether mapping was possible or not. CONCLUSIONS: The dissection revealed that several nursing diagnostic concepts can easily be interchanged, while others cannot or can be mapped only for specific purposes (e.g., clinical or aggregate use). This implies that for some concepts it does not matter which terminology is used, and in other cases it does because of different meanings. PMID- 17117946 TI - Stress overload: a new diagnosis. AB - PURPOSE: To describe the phenomenon of stress overload as a nursing diagnosis. METHODS: A qualitative study using case study method was conducted with nine adults experiencing stress overload to fully describe the experience and identify possible defining characteristics. Current literature sources on stress and its related factors were examined to support stress overload as a nursing diagnosis for inclusion in the NANDA International classification. FINDINGS: Stress overload, defined as excessive amounts and types of demands that require action, is a human response that is experienced as a problem and contributes to the development of other problems. The proposed defining characteristics are perceives situational stress as excessive, expresses a feeling of tension or pressure, expresses difficulty in functioning as usual, expresses problems with decision-making, demonstrates increased feelings of anger and impatience, and reports negative effects from stress such as physical symptoms or psychological distress. PRACTICE IMPLICATIONS: Nursing interventions such as active listening and decision-making support are needed to help people reduce stress levels. Studies are needed to further validate the defining characteristics and related factors of this new diagnosis. PMID- 17117947 TI - Investigation of the functional variant c.-169T > C of the Fc receptor-like 3 (FCRL3) gene in alopecia areata. AB - A functional variant in the Fc receptor-like 3 (FCRL3) gene has been implicated in susceptibility to autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus and autoimmune thyroid disease. Investigating a large case control sample of patients with alopecia areata (AA), we found no evidence for the involvement of FCRL3 in susceptibility to AA. PMID- 17117948 TI - Sequencing-based analysis of the HLA-DPB1 polymorphism in Nu ethnic group of south-west China. AB - In the present study, DNA typing for HLA-DPB1 was performed using polymerase chain reaction-sequencing-based typing method in 72 randomly selected Nu ethnic individuals inhabiting the Yunnan province of south-west China. Among the 12 detected DPB1 alleles, the most frequent was DPB1*1301, with the percentage of 20.83%, followed by DPB1*0501 (19.44%), DPB1*040101 (16.67%) and DPB1*2801 (9.72%). The allele DPB1*1501 was found for the first time in the Chinese population. Neighbour-joining showed that the Nu ethnic minority belonged to East Asian cluster and was most closely related to Lisu, being consistent with the historical records. In addition, the results obtained in this study will also provide useful information on organ transplantation, forensic investigations and disease association studies. PMID- 17117949 TI - Association screen for atopic dermatitis candidate gene regions using microsatellite markers in pooled DNA samples. AB - Atopic dermatitis (AD) is a chronic inflammatory skin disease affecting up to 16% of children in developed countries. A complex genetic background for AD has been suggested, with genetic as well as environmental factors influencing disease susceptibility. Among other factors, dysregulation in both the innate and the adaptive immune system has been proposed to play a role in AD pathophysiology. We present here an extended association screen for AD using microsatellite markers in 154 genes related to innate and adaptive immunity in pooled DNA samples from 150 German children with AD and 100 controls. After Bonferroni correction, no marker revealed a significant association with AD. Yet, markers representing the nuclear factor kappa B (NFKB)1 and chemokine receptor (CCR)4 genes showed differences in allelic distributions between cases and controls for both pooled DNA analysis and individual genotyping and were thus further investigated. Evaluation of additional single nucleotide polymorphisms (SNP) in the NFKB1 and CCR4 genes revealed no association of individual SNPs with AD. In contrast, haplotype analyses showed a significantly different haplotype distribution between patients and controls for CCR4 (P < 0.001). Furthermore, when SNP-SNP interaction effects were analysed for these two genes, we found significant evidence for epistatic interactions between SNPs within each of the two genes but no evidence for a gene-gene interaction, suggesting that variation in or near both the CCR4 and the NFKB1 genes might individually contribute to AD pathogenesis. PMID- 17117950 TI - Association of -308 TNF-alpha promoter polymorphism with type 1 diabetes in North Indians. PMID- 17117951 TI - Absence of association between two insertion/deletion coding genetic polymorphisms of TIM-1 gene and asthma in Chinese Han population. AB - TIM-1, a member of T-cell immunoglobulin domain and mucin domain (TIM) gene family, was implicated as an asthma susceptibility gene in previous studies. TIM 1 was expressed on CD4(+) T cells after activation and its expression was sustained preferentially in T-helper type 2 (T(H)2) but not in T(H)1 cells, therefore TIM-1 became a good candidate gene for atopic diseases. Recent studies indicated that two insertion/deletion (ins/del) coding genetic polymorphisms in exon 4 of TIM-1 were associated with asthma susceptibility in some but not in all populations. In order to investigate the relationship between TIM-1 genetic polymorphisms and asthma in Chinese Han population, we performed a case-control study for two insertion/deletion polymorphisms in TIM-1 exon 4 (5383_5397ins/del and 5509_5511delCAA) and a single nucleotide polymorphism (SNP) in intron 8 (IVS 8+9 G/A) between a healthy control group of 309 people and an asthma patient group of 352 people recruited from Chinese Han population. The polymorphisms were genotyped and the allele and genotype frequencies were analysed, but none of the three polymorphisms showed association with asthma susceptibility in single-locus association analyses. Linkage disequilibrium (LD) analyses demonstrated that the two insertion/deletion polymorphisms were in strong LD but the haplotypes constructed from these two polymorphisms showed no significant association with asthma. In conclusion, our findings suggest that 5383_5397ins/del, 5509_5511delCAA and SNP IVS 8+9 G/A polymorphisms are not associated with asthma susceptibility in Chinese Han population. PMID- 17117952 TI - Upregulating promoter polymorphisms of RANTES relate to atopic dermatitis. AB - It has been reported that a functional polymorphism in the promoter of the RANTES gene (-403G/A) is associated with atopic dermatitis in a German population. Although there are several reports on the association of RANTES promoter polymorphisms (-403G/A and -28C/G) with asthma, the association of these polymorphisms with atopic dermatitis has not yet been confirmed in other populations. We therefore aimed to test whether the RANTES promoter polymorphisms relate to atopic dermatitis in a well-defined Japanese population. We conducted an association study of upregulating promoter polymorphisms of RANTES (-403G/A and -28C/G) in 389 patients with atopic dermatitis and 177 healthy control subjects. There was a significant association between the upregulating variant of RANTES -28G and atopic dermatitis, while -403A variant showed a significant association with atopic dermatitis with high IgE productivity. These results support a role for RANTES promoter polymorphisms in susceptibility to atopic dermatitis. PMID- 17117953 TI - Th1 and Th2 cytokine gene polymorphisms in two indigenous ethnic groups in Iran. AB - Cytokines are important immunomodulatory molecules in immune responses against microorganisms and also have an important role in the setting of disorders affecting immune system. Cytokine single nucleotide polymorphisms have been extensively studied in different normal populations as well as in relation to diseases. Some of these polymorphisms (SNP) affect cytokine gene transcription and expression. The polymorphisms of cytokine genes are potentially important as genetic predictors of the disease susceptibility and clinical outcome or as a tool for anthropological studies. In this study, samples have been collected from 261 healthy individuals located in two different regions of Iran (Tehran and Yazd). The allele and genotype frequencies of Th1 and Th2 cytokines SNP including interleukin (IL)-2, IL-4, IL-6, IL-10, tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma have been investigated, using the polymerase chain reaction-sequence-specific primer method. The allele and genotype frequencies in Tehran and Yazd populations were similar except for the IL-2, IL-4, IL-6 and IL 10. The IL-4 C allele, C/C -33 and T/T -1098 genotype were significantly more frequent in Tehran than in Yazd population (P = 0.04, P = 0.004 and P = 0.001, respectively). The G/A genotype of the IL-6 (nt565) and IL-10 (-1082) was significantly less frequent in Tehran than in Yazd population (P = 0.01 and P = 0.003, respectively). The GT haplotype of the IL-2 (-330, +166) was significantly less frequent in Tehran than in Yazd population (P = 0.0002). We have also compared our whole samples with the reported data from other countries showing that Iranian population have cytokine gene polymorphism profile similar to that of Caucasians, especially Italian population. PMID- 17117954 TI - Allelic distribution of HLA class I genes in the Tibetan ethnic population of China. AB - Tibetans live in Qinghai-Tibet Plateau rising about 4000 m a.s.l. in south-west China. Archaeological evidences suggested that there have been humans living in Tibet at least 5000 years ago. However, Tibetan earlier history remains elusive. In the present study, allelic distribution of human leucocyte antigen (HLA)-A, -B and -Cw in 158 unrelated Tibetan Chinese was investigated using sequencing-based typing methods, and a total of 25 HLA-A, 45 HLA-B and 20 HLA-Cw alleles were identified. A*24G1 (27.2%), B*51G1 (16.8%), Cw*04G1 (13.3%) and Cw*070201G1 (13.3%) are the most common HLA-A, -B and -Cw alleles. The most frequently detected haplotypes were A*24G1-B*51G1-Cw*140201 (3.6%), A*24G1-B*51G1 (6.8%), A*02G1-Cw*070201G1 (6.5%) and B*51G1-Cw*140201 (5.0%). Chi-squared test suggested that all three loci fitted the Hardy-Weinberg expectations. No evidence for a departure from selective neutrality at the HLA-A and -B loci was observed. However, significant departure of the observed homozygosity from the expected values was found for HLA-Cw. Though the contemporary Tibetans inhabit the south west China, Nei's genetic distance measure based on frequencies of HLA-A, -B and Cw indicated that Tibetans were closer to northern Han Chinese, Mongolian Chinese, Koreans and Japanese rather than to southern Han Chinese. The corresponding dendrogram constructed according to the neighbour-joining method supported that Tibetans separated from southern Han and located in North-East Asian cluster which included northern Han Chinese and Mongolian Chinese. These data were in good agreement with language classification and with a recent hypothesis that Tibetan might originate from northern China along Yellow river. PMID- 17117959 TI - Ethanol-related behaviors in serotonin transporter knockout mice. AB - BACKGROUND: Increasing evidence supports a role for 5-hydroxytryptamine (5-HT) and the 5-HT transporter (5-HTT) in modulating the neural and behavioral actions of ethanol (EtOH) and other drugs of abuse. METHODS: We used a 5-HTT knockout (KO) mouse model to further study this relationship. 5-Hydroxytryptamine transporter KO mice were tested for the sedative/hypnotic, hypothermia-inducing, motor-incoordinating (via accelerating rotarod), and depression-related (via tail suspension test) effects of acute EtOH administration. Reward-related effects of EtOH were assessed in 5-HTT KO mice using the conditioned place preference (CPP) paradigm. 5-Hydroxytryptamine transporter KO mice were tested for voluntary consumption of EtOH in a modified 2-bottle choice test that measured the temporal organization of drinking over the circadian cycle via "lickometers." RESULTS: Replicating previous findings, 5-HTT KO mice exhibited significantly increased sensitivity to EtOH-induced sedation/hypnosis relative to wild-type controls. Additionally, 5-HTT KO mice showed motor-coordination deficits at baseline and in response to EtOH. Hypothermic, pro-depressive-like, and reward-related effects of EtOH were no different across genotypes. Gross EtOH consumption was modestly reduced in 5-HTT KO mice, due to significantly lesser consumption during the peak period of drinking in the early dark phase. CONCLUSIONS: Data extend the finding that loss of 5-HTT gene function alters certain neural and behavioral effects of EtOH, with implications for better understanding the pathophysiology and treatment of alcoholism. PMID- 17117960 TI - Methylenetetrahydrofolate reductase C677T-polymorphism and its association with alcohol withdrawal seizure. AB - BACKGROUND: Elevated homocysteine plasma levels are considered as a risk factor for the occurrence of seizures during alcohol withdrawal. Homocysteine plasma concentrations seem to be influenced by the methylenetetrahydrofolate reductase (MTHFR) C677T-polymorphism. It was investigated whether the T-allele of the MTHFR C677T-polymorphism is associated with alcohol dependence, alcohol withdrawal seizure (WS), or the daily amount of alcohol consumption. METHODS: A group of 102 healthy controls and 221 alcoholic patients, including 97 patients with a history of mild withdrawal symptoms (MWS) and 70 patients with a history of alcohol WS, were genotyped, and personal data were collected for statistical evaluation in a case-control design. RESULTS: The T-allele is significantly associated with WS by comparing alcoholic patients with a history of WS (T-allele frequency: 0.39) and healthy controls (T-allele frequency: 0.28) (p=0.03). Although there was no significant difference between alcoholic patients with only MWS and alcoholic patients with a history of WS, a trend for the T-allele frequency among the analyzed subgroups was noticed: T-allele frequency increased from f(T)=0.28 in healthy controls to f(T)=0.33 in alcoholic patients with MWS up to f(T)=0.40 in alcohol-dependent men having a WS. Differences between healthy male controls and male alcoholic patients concerning the T-allele frequency also turned out to be significant [f(T)=0.27 vs f(T)=0.37; p=0.03]. Daily alcohol intake was independent of T-allele carrier status in alcohol-dependent patients. CONCLUSION: The present study suggests an influence of the MTHFR C677T-polymorphism on the etiology of alcohol WS and alcohol dependence in men in a western European population. An influence of MTHFR C677T on the daily amount of alcohol intake before admission among alcohol-dependent patients could not be shown. PMID- 17117961 TI - A joint genomewide linkage analysis of symptoms of alcohol dependence and conduct disorder. AB - BACKGROUND: A large linkage peak for alcohol dependence (AD) was detected on chromosome 4q in the Irish Affected Sib Pair Study of Alcohol Dependence (IASPSAD). Are the susceptibility genes underlying this peak specific for AD or do they increase risk for externalizing disorders more generally? Can we, in the IASPSAD, replicate prior evidence for linkage to conduct disorder (CD)? METHODS: The 733 all possible sibling pairs in IASPSAD were typed for 1,020 short-tandem repeat genetic markers. Univariate and bivariate linkage analyses were conducted by the program sequential oligogenic linkage analysis routines (SOLAR), for both the raw and the transformed number of symptoms of AD (ADsx) and number of symptoms of CD (CDsx). In the bivariate analyses, specificity was assessed by the ratio of the variance accounted for in ADsx and CDsx by the quantitative trait locus. RESULTS: In the univariate linkage analyses, no evidence for linkage to CDsx was found under the 4q peak for ADsx and the largest peaks for CDsx were seen on chromosomes 1q (LOD=3.16) and 14p (LOD=2.36). In the bivariate linkage analysis, the 4q peak had high specificity for AD (AD/CD ratio of 39.9). Several smaller peaks, on chromosomes 1, 7, and 10, had moderate specificity for CD but also impacted on risk for AD, with AD/CD ratios of 0.18 to 0.32. CONCLUSIONS: Genes under the 4q linkage peak for AD in the IASPSAD impact specifically on risk for AD rather than more broadly on risk for externalizing syndromes. Suggestive linkages were found in several locations for CD, 2 of which broadly replicate prior findings. The bivariate analyses identified genomic locations containing susceptibility loci that impacted on risk for both CDsx and ADsx. PMID- 17117962 TI - Ethanol-induced regulation of GABA-A subunit mRNAs in prefrontal fields of cynomolgus monkeys. AB - BACKGROUND: Recent evidence indicates that functional impairment of the orbital and medial fields of the prefrontal cortex may underlie the deficits in executive control of behavior that characterize addictive disorders, including alcohol addiction. Moreover, previous studies have indicated that alcohol alters GABA neurotransmission and one substrate of these effects may be through the reconfiguration of the subunits constituting the GABA(A) receptor complex. Given that GABAergic transmission has an integral role in cortical processing, influencing local and interregional communication, understanding alcohol-induced alterations in GABA(A) receptors in prefrontal fields of the primate brain may provide insight into the functional impairment of these brain regions in the alcohol-addicted state and extend our understanding of the molecular consequences of long-term use in these critical brain regions. METHODS AND RESULTS: To address this problem, the effects of chronic ethanol self-administration in male cynomolgus monkeys on GABA(A) receptor subunit mRNA expression was studied in 3 frontal cortical fields: orbitofrontal cortex (OFC; area 13), anterior cingulate cortex (ACC; area 24), and the dorsolateral prefrontal cortex (DLPFC; area 46). Quantitative polymerase chain reaction revealed significant alterations in GABA(A) subunit mRNA expression in the OFC and DLPFC but not in the ACC. Specifically, expression of the alpha2, alpha4, beta1, beta3, and gamma1 to gamma3 subunit mRNAs was significantly less in the OFC, whereas the expression of beta1, beta2, gamma1, and delta subunit mRNAs was less in the DLPFC of alcohol treated monkeys. CONCLUSION: These findings suggest that ethanol-induced alterations in GABA(A) function may be due to alterations in GABA(A) subunit mRNA levels and subunit-specific alterations are selective to particular cortical fields. PMID- 17117963 TI - Patterns of regional brain activity in alcohol-dependent subjects. AB - BACKGROUND: Electroencephalographic (EEG) measures of hemispheric asymmetry in anterior brain activity have been related to a variety of indices of psychopathology and emotionality. However, little is known about patterns of frontal asymmetry in alcohol-dependent (AD) samples. It is also unclear whether psychiatric comorbidity in AD subjects accounts for additional variance in frontal asymmetry, beyond a diagnosis of AD alone. METHODS: We compared 193 AD subjects with 108 control subjects on resting brain activity in anterior and posterior regions, as indexed by asymmetries in alpha band power in the left and right hemispheres. Within the AD group alone, we examined whether comorbid major depressive disorder (MDD) or antisocial personality disorder (ASPD) had effects on regional asymmetry. RESULTS: Compared with control subjects, AD subjects exhibited lower left, relative to right, cortical activation in anterior regions. Evidence that comorbidity in AD subjects accounted for further variance in EEG asymmetry was mixed; AD subjects with comorbid ASPD were not significantly different from those without ASPD, while AD subjects with a lifetime history of MDD showed less asymmetry in anterior regions than those without MDD. CONCLUSIONS: Our findings indicate that AD subjects exhibit a pattern of frontal asymmetry similar to that found in other psychiatric groups. Results examining the effects of comorbidity in AD on EEG asymmetry were inconclusive. The implications of our findings for future work are described. PMID- 17117964 TI - Perception of sleep in recovering alcohol-dependent patients with insomnia: relationship with future drinking. AB - BACKGROUND: Subjective and objective measures of poor sleep in alcoholic insomniacs predict relapse to drinking. Nonalcoholic insomniacs underestimate their total sleep time (TST) and overestimate their sleep onset latency (SOL) and wake time after sleep onset (WASO) compared with polysomnography (PSG). This study evaluated 3 hypotheses: (1) subjective SOL would predict frequency of future drinking; (2) participants would overestimate SOL and WASO and underestimate TST; and (3) higher amounts of over- and underestimates of sleep at baseline would predict worse drinking outcomes prospectively. METHODS: Participants (N=18), mean age 44.6 years (+/-13.2), underwent an adaptation night and then 2 nights of PSG 3 weeks apart. They also provided morning estimates of SOL, WASO, TST, and sleep efficiency (SE). Following the baseline PSG, participants were followed over 12 weeks. A 2-way ANOVA (night x method of measuring sleep) compared results and regression analyses predicted drinking. Drinking outcomes were defined as number of days drinking (DD) and number of heavy-drinking days (HDD) during 2 consecutive 6-week follow-up periods. RESULTS: Most participants (72%) overestimated SOL by a mean of 21.3 (+/-36) minutes compared with PSG [F(1, 14)=7.1, p<0.03]. Unexpectedly, 89% underestimated WASO by a mean difference of 48.7 (+/-49) minutes [F(1, 14)=15.6, p<0.01]. Drinking during the first 6-week study period was predicted by both subjective estimates of WASO and their accuracy, whereas drinking during the second 6-week period was predicted by both subjective estimations of sleep and rapid eye movement sleep latency. CONCLUSION: Greater subjective accuracy of wakefulness at night provided by the patient predicted drinking during the study. Unlike nonalcoholic insomniacs, this alcoholic sample significantly underestimated WASO compared with PSG values. The predictive ability of sleep parameters depended on the selected measure of drinking outcomes and when outcomes were measured. Subjective sleep measures were better predictors of future drinking than corresponding PSG measures. PMID- 17117965 TI - Visual P300s in long-term abstinent chronic alcoholics. AB - BACKGROUND: Evidence of reduced P3b amplitudes in chronic alcoholics and individuals at risk for developing alcoholism suggest that the P3b may be an endophenotypic marker for alcoholism. If this is the case, then long-term abstinent alcoholics (LTAAs) should exhibit reduced P3b amplitudes. Thus far, P3b studies on chronic alcoholics have focused primarily on samples with relatively short-term abstinence (less than 15 months). This study examines the amplitude and latency of the P3b and P3a event-related brain electrical components in LTAAs compared with normal controls (NCs) and whether these measures are related to alcohol use and other subject variables. METHODS: Electroencephalographs (EEGs) were recorded on 48 LTAAs (mean abstinence=6.7 years) compared with 48 age matched and gender-matched NCs during a visual P300 experiment consisting of standard, target, and rare nontarget conditions. This paradigm elicited the P3b (target condition) and the P3a (rare nontarget condition) components. RESULTS: Long-term abstinent alcoholics had reduced P3b amplitudes and increased P3b latencies in comparison with NCs. Long-term abstinent alcoholics also exhibited delayed P3a components, but no P3a amplitude reductions. Alcohol use variables, a family history of alcohol problems, and the duration of alcohol abstinence were not associated with any amplitude or latency variables. CONCLUSIONS: Even after very prolonged abstinence, reduced P3b amplitudes are present in chronic alcoholics and are not associated with any family history or alcohol use variables. These results provide equivocal support for reduced P3b amplitude being an endophenotypic marker for alcoholism, but are also consistent with P3b being affected by a threshold of alcohol abuse, with the effect not resolving over long periods of abstinence. PMID- 17117966 TI - Time-dependent alterations in ethanol intake in male wistar rats exposed to short and prolonged daily maternal separation in a 4-bottle free-choice paradigm. AB - BACKGROUND: Previous studies have shown that maternal separation can be used in animal studies of early environmental influence on adult ethanol intake. These studies have shown that short daily separations result in low ethanol intake, whereas prolonged separations relate to an enhanced risk for a high ethanol intake. The aim of the present study was to further examine the long-term effects of early-life events on ethanol intake. METHODS: Rat pups were exposed to 15 minutes (MS15) or 360 minutes (MS360) of daily maternal separation during postnatal days 1 to 21 or kept under normal animal facility rearing (AFR) conditions. In adulthood, male rats were given free access to 5, 10, and 20% ethanol, in addition to water, in a 4-bottle-choice paradigm. RESULTS: No differences in total ethanol intake or preference between the 3 experimental groups were found. The 54-day drinking period was divided into acquisition, stabilization, and maintenance phases for analysis of time and group differences. The MS15 rats increased ethanol intake over time; they mostly consumed 5% ethanol and had a low intake of 20% ethanol throughout the experiment. MS360 rats increased ethanol intake, changed preference from 5% to 20% ethanol, and had a higher increase in intake of 20% ethanol over time. The ethanol intake and preference in the AFR rats were more similar to that of the MS360 rats. CONCLUSIONS: Time-dependent changes were observed in the preferred choice of low versus high ethanol concentrations in MS15 and MS360 rats. The results support previous findings suggesting that MS15 can be used as a model for environmental protective factors and that MS360 represents a risk environment for acquisition of a high adult ethanol intake. PMID- 17117967 TI - Development of an alcohol deprivation and escalation effect in C57BL/6J mice. AB - BACKGROUND: Relapse-like drinking has been studied through the expression of the alcohol deprivation effect (ADE), which is measured by a pronounced increase in ethanol preference and consumption after imposed abstinence. No studies have characterized the ADE in C57BL/6J (B6) mice. The present study examined the effects of length and number of deprivations on the expression of the ADE in B6 mice. METHODS: Adult male B6 mice received 24-hour continuous access to ethanol and water for 6 weeks (baseline). Experiment 1 determined the ADE in mice receiving weekly access to 15% ethanol (i.e., exposed 1 day a week and deprived during the other 6 days) for a total of 10 weeks. Experiments 2 and 3 determined the ADE after a single 2-week deprivation period in mice receiving a single concentration of 15% ethanol or multiple concentrations of 7.5, 15, and 30% ethanol, respectively, followed by weekly access to their respective ethanol solutions for 10 weeks. Experiment 4 determined the ADE after a single 2-week deprivation period, followed by daily access to 15% ethanol. Mice never deprived of ethanol (i.e., continuous access) were used as age-matched drinking controls. RESULTS: The ADE was observed after the initial 6-day deprivation period and was profoundly enhanced (i.e., escalation of the ADE) following weekly reexposure to 15% ethanol. Compared with a single concentration of 15% ethanol, concurrent access to multiple ethanol concentrations resulted in a near 2-fold increase in baseline ethanol consumption. Regardless of having access to single or multiple concentrations of ethanol, the ADE was not observed immediately after a 2-week deprivation period. The ADE was observed (although to a lesser magnitude and duration) following weekly reexposure to single or multiple concentrations of ethanol. Alternatively, following a 2-week deprivation period, mice receiving daily access to 15% ethanol showed a significant decrease in ethanol intake and preference (i.e., negative ADE). CONCLUSIONS: Short-term deprivations followed by repeated intermittent (weekly) reexposure to ethanol produces a robust ADE in B6 mice. Increasing the initial deprivation length to 2 weeks produces various opposing effects, including erasure of an initial ADE, diminished expression and magnitude of the ADE following weekly exposure, and complete reversal of the ADE following daily exposure to ethanol. PMID- 17117968 TI - Environmental cues, alcohol seeking, and consumption in baboons: effects of response requirement and duration of alcohol abstinence. AB - BACKGROUND: Environmental stimuli (cues) that have been paired with alcohol drinking may evoke classically conditioned states that in turn influence alcohol consumption and relapse to heavy drinking. Animal models using chained schedules of alcohol reinforcement may be useful for examining such complex interactions. METHODS: Alcohol drinking was established in 4 baboons. A sequence of lights and tones was presented during daily 3-hour sessions. First, cues were presented alone and no programmed contingencies were in effect. Second, cues were paired with 3 linked components consisting of different behavioral contingencies leading to and concluding with access to alcohol for self-administration in the last component (i.e., a chained schedule of alcohol reinforcement). Third, the effects of withholding alcohol access (i.e., forced abstinence) and increasing the number of lever responses required per drink were evaluated. RESULTS: Cues paired with a chained schedule of alcohol reinforcement engendered behaviors that brought baboons into contact with alcohol-related cues and occasioned operant responding that facilitated access to alcohol (alcohol seeking) during components that preceded alcohol access. Increasing the response requirement for each drink decreased the number of drinks and volume of alcohol consumed, but did not alter alcohol seeking. On the first session after 14 days of alcohol abstinence, latency to complete the operant requirement that produced alcohol access was decreased while both alcohol self-administration and volume of alcohol consumed were increased. CONCLUSIONS: Alcohol self-administration and consumption were sensitive to increases in response requirement and duration of alcohol abstinence, while seeking was only enhanced by duration of alcohol abstinence. This animal model may be useful to further examine the interactions between environmental cues and behaviors associated with seeking and consumption of alcohol and to evaluate the efficacy of potential alcohol treatment drugs on these behaviors. PMID- 17117969 TI - Comparison of motor delays in young children with fetal alcohol syndrome to those with prenatal alcohol exposure and with no prenatal alcohol exposure. AB - BACKGROUND: Researchers are increasingly considering the importance of motor functioning of children with fetal alcohol spectrum disorder (FASD). The purpose of this study was to assess the motor development of young children with fetal alcohol syndrome (FAS) to determine the presence and degree of delay in their motor skills and to compare their motor development with that of matched children without FAS. METHODS: The motor development of 14 children ages 20 to 68 months identified with FAS was assessed using the Vineland Adaptive Behavior Scales (VABS). In addition, 2 comparison groups were utilized. Eleven of the children with FAS were matched for chronological age, gender, ethnicity, and communication age to: (1) 11 children with prenatal alcohol exposure who did not have FAS and (2) 11 matched children without any reported prenatal alcohol exposure. The motor scores on the VABS were compared among the 3 groups. RESULTS: Most of the young children with FAS in this study showed clinically important delays in their motor development as measured on the VABS Motor Domain, and their fine motor skills were significantly more delayed than their gross motor skills. In the group comparisons, the young children with FAS had significantly lower Motor Domain standard (MotorSS) scores than the children not exposed to alcohol prenatally. They also had significantly lower Fine Motor Developmental Quotients than the children in both the other groups. No significant group differences were found in gross motor scores. For MotorSS scores and Fine Motor Developmental Quotients, the means and standard errors indicated a continuum in the scores from FAS to prenatal alcohol exposure to nonexposure. CONCLUSIONS: These findings strongly suggest that all young children with FAS should receive complete developmental evaluations that include assessment of their motor functioning, to identify problem areas and provide access to developmental intervention programs that target deficit areas such as fine motor skills. Fine motor delays in children with FAS may be related to specific neurobehavioral deficits that affect fine motor skills. The findings support the concept of an FASD continuum in some areas of motor development. PMID- 17117970 TI - Cigarette smoking and the risk for alcohol use disorders among adolescent drinkers. AB - BACKGROUND: Cigarette smoking and alcohol use disorders (AUDs) are closely linked, but it is not clear whether higher rates of AUD among smokers are solely attributable to heavier drinking or, alternatively, whether smokers are more vulnerable to alcohol abuse and dependence than nonsmokers who drink comparable quantities. We sought to address this issue using data from a nationally representative U.S. sample of adolescents and young adults. Specifically, we analyzed the relationship between cigarette smoking, drinking, and AUDs. METHODS: Data were from the aggregated 2002 through 2004 U.S. National Survey on Drug Use and Health. Participants were randomly selected, household-dwelling adolescents and young adults (ages 12-20) from the noninstitutionalized, civilian population of the United States (N=74,836). Measurements included current DSM-IV alcohol abuse or dependence, number of drinks in the past 30 days, and past-year cigarette smoking, defined as having smoked more than 100 cigarettes across the lifetime and having smoked during the past year. RESULTS: Past-year smokers (prevalence=16.0%) drank in higher quantities than never-smokers, but were also at elevated risk for AUD when compared with never-smokers who drank equivalent quantities. The effect was observed across age groups, but was more prominent among younger adolescents. After adjusting for drinking quantity and sociodemographic variables, smokers had 4.5-fold higher odds of AUD than never smokers [95% confidence interval (95% CI), 3.1-6.6]. Youths who reported smoking but did not cross the 100-cigarette threshold were at intermediate risk [odds ratio (OR), 2.3; 95% CI, 1.7-3.3]. Differences in AUD between smokers and never smokers were most pronounced at lower levels of drinking. CONCLUSIONS: The results are consistent with a higher vulnerability to AUDs among smokers, compared with nonsmokers who drink equivalent quantities. PMID- 17117971 TI - Infant stress reactivity and prenatal alcohol exposure. AB - BACKGROUND: Animal studies have shown that prenatal alcohol exposure (PAE) is linked to alterations in the stress response systems. To date, little is known about the impact of PAE on stress systems in human infants. The current study examined PAE effects on the stress response, as evidenced by the activation of the limbic-hypothalamic-pituitary-adrenal (L-HPA) axis and autonomic system and changes in negative affect during a social-emotional challenge in human infants. We also examined whether the effects of PAE on infant responsiveness differed in boys and girls. METHODS: Measures of cortisol, heart rate, and negative affect were obtained during a modified version of Tronick's still-face procedure, a standardized developmental paradigm used to study emotion and stress regulation. Our sample included fifty-five 5- to 7-month-old infants whose mothers were enrolled in an alcohol intervention study. Measures of maternal alcohol consumption during pregnancy and after delivery were obtained using Timeline Followback interviewing methods. Relationships between prenatal alcohol consumption and infant outcomes were examined. In addition, mothers were divided into high and low-frequency drinkers, based on the mean percent of prenatal drinking days (PDD) to facilitate between-group comparisons of infant stress measures. RESULTS: Mothers enrolled in our study reported significant reductions in alcohol consumption after learning of their pregnancies. Nevertheless, PDD from conception to pregnancy recognition was related to increases in cortisol reactivity, elevated heart rate, and negative affect in their infants. The effects of PAE on infant responsiveness were significant after controlling for the effects of maternal depression and annual income. In addition, the effects of PAE on cortisol reactivity differed for boys and girls. CONCLUSIONS: Greater PAE was related to greater activation of stress response systems. Our findings suggest that PAE affects the development of infant stress systems and that these effects differ in boys and girls. This work supports the possibility that PAE is related to alterations in infant stress systems, which could underlie problems in cognitive and social-emotional functioning that are common among persons exposed prenatally to alcohol. PMID- 17117972 TI - Chronic alcohol accentuates nutritional, metabolic, and immune alterations during asymptomatic simian immunodeficiency virus infection. AB - BACKGROUND: Alcohol abuse has been reported to have a high prevalence in the human immunodeficiency virus (HIV)-infected population. However, its impact on disease progression is unknown. Studies dissecting the drug-induced or alcohol induced metabolic derangements that are likely to alter the course of disease progression are lacking. This is particularly important because of the substantial reduction in morbidity and mortality of patients on highly active antiretroviral therapy (HAART). HIV infection has become a more chronic disease during which alcohol-induced metabolic alterations may become more prevalent and pronounced. METHODS: The present study used a model of chronic intragastric alcohol administration initiated 3 months before intravenous simian immunodeficiency (SIV) inoculation and continued thereafter throughout the course of SIV infection, to investigate the impact of chronic alcohol binge-like consumption during the initial 10-month asymptomatic phase of SIV infection in nonhuman primate rhesus macaques. Anthropometric, metabolic, biochemical, nutritional, and immune state indicators were examined before infection and at 3 month intervals in asymptomatic chronic alcohol-treated SIV-infected macaques and time-matched isocaloric and uninfected controls. RESULTS: Intravenous SIV(DeltaB670) infection resulted in increased viral load, decreased circulating CD4(+)/CD8(+) lymphocyte ratio, and increased lymphocyte proliferation (Ki67/CD3(+)). Chronic alcohol/SIV(+) animals showed a higher viral load at 3 months post-SIV infection as well as a significant and early decrease in caloric intake and nitrogen balance associated with a change in food choice. Rates of skeletal muscle protein synthesis and breakdown, mRNA expression of IGF-I, myostatin, or the ubiquitin ligase muscle atrophy F-box protein (MAFbx) did not differ from basal during the 10-month asymptomatic period of infection. However, muscle TNF-alpha mRNA expression was markedly increased at 10 months post-SIV infection in alcohol/SIV(+) animals. DISCUSSION: These findings suggest that chronic alcohol accelerates nutritional and metabolic dysregulation during SIV infection and may favor a skeletal muscle proinflammatory state, possibly conducive to subsequent muscle wasting. PMID- 17117973 TI - Activation and impairment of platelet function in vitro by fatty acid ethyl ester, a nonoxidative ethanol metabolite: effect of fatty acid ethyl esters on human platelets. AB - BACKGROUND: The goal of this study was to investigate the effect of fatty acid ethyl esters (FAEE), nonoxidative metabolites of ethanol, on platelet function. We hypothesized that FAEE increase the risk of bleeding by producing an alteration in platelet membrane structure or function. METHODS: Isolated human platelets incubated with FAEE were prepared and multiple assays for platelet activation were performed; beta-thromboglobulin release from platelet granules, platelet aggregation, arachidonate release from phospholipids, and intracellular cyclic AMP (cAMP) levels. We examined also the combined effect of epinephrine and FAEE on platelet aggregation. RESULTS: FAEE induced platelet shape change, release of alpha granules and release of arachidonate from phospholipids without an increase in eicosanoid production and decreased cAMP levels. The platelets did not aggregate in response to FAEE alone, but did shorten the time to maximum aggregation with epinephrine. CONCLUSION: These studies show that FAEE potentiate platelet activation but do not induce aggregation, presumably because they do not stimulate thromboxane A(2) production. PMID- 17117974 TI - Reinforcing effects of central ethanol injections in newborn rat pups. AB - BACKGROUND: Newborn rat pups readily ingest ethanol of low to moderate concentrations and are sensitive to its reinforcing effects. Given that early ethanol exposure can promote its future abuse, it is vital to discover the mechanisms behind reinforcing effects of ethanol at this stage of life. METHODS: Cesarean-delivered 3- to 4-hour-old rat pups were exposed to lemon odor (unconditioned stimulus) either paired or explicitly unpaired with central injections of saline or ethanol (25, 50, 100, 200, or 400 mg%) in a volume of 1 muL. One hour following conditioning subjects were tested on a surrogate nipple providing water in the presence of lemon odor. Reinforcement from ethanol's central effects was indexed by significantly greater attachment time on a lemon scented nipple in paired subjects than in unpaired or saline controls. RESULTS: Rats centrally injected with 25 to 200 mg% ethanol in the presence of lemon odor spent significantly more time attached to a lemon-scented surrogate nipple providing water than did their saline-injected or unpaired counterparts. Those injected with 400 mg% ethanol did not differ from their corresponding controls. No detectable brain alcohol content was found in the assay of whole brain for ethanol levels. CONCLUSIONS: These results indicate that 3- to 4-hour-old rat pups find central injections of 25 to 200 mg% ethanol reinforcing. This procedure virtually eliminates ethanol's chemosensory or caloric attributes as the source of ethanol reinforcement. The present classical olfactory conditioning paradigm can be used to further study mechanisms of this apparently pharmacological reinforcement by ethanol in newborn rat pups. PMID- 17117975 TI - Brain metabolic alterations in adolescents and young adults with fetal alcohol spectrum disorders. AB - BACKGROUND: Prenatal alcohol exposure affects brain structure and function. This study examined brain metabolism using magnetic resonance spectroscopy (MRS) and searched for regions of specific vulnerability in adolescents and young adults prenatally exposed to alcohol. METHODS: Ten adolescents and young adults with confirmed heavy prenatal alcohol exposure and a diagnosis within the fetal alcohol spectrum disorders (FASD) were included. Three of them had fetal alcohol syndrome (FAS), 3 had partial FAS (PFAS), and 4 had alcohol-related neurobehavioral disorder (ARND). The control group consisted of 10 adolescents matched for age, sex, head circumference, handedness, and body mass. Exclusionary criteria were learning disorders and prenatal alcohol exposure. Three-dimensional (1)H magnetic resonance spectroscopic imaging ((1)H MRSI) was performed in the cerebrum and cerebellum. Metabolite ratios N-acetylaspartate/choline (NAA/Cho), NAA/creatine (Cr) and Cho/Cr, and absolute metabolite intensities were calculated for several anatomic regions. RESULTS: In patients with FASD, lower NAA/Cho and/or NAA/Cr compared with controls were found in parietal and frontal cortices, frontal white matter, corpus callosum, thalamus, and cerebellar dentate nucleus. There was an increase in the absolute intensity of the glial markers Cho and Cr but no change in the neuronal marker NAA. CONCLUSIONS: Our results suggest that prenatal alcohol exposure alters brain metabolism in a long-standing or permanent manner in multiple brain areas. These changes are in accordance with previous findings from structural and functional studies. Metabolic alterations represent changes in the glial cell pool rather than in the neurons. PMID- 17117976 TI - Bacterial competition between a bacteriocin-producing and a bacteriocin-negative strain of Streptococcus bovis in batch and continuous culture. AB - A bacteriocin-producing Streptococcus bovis strain (HC5) outcompeted a sensitive strain (JB1) before it reached stationary phase (pH 6.4), even though it grew 10% slower and cell-free bovicin HC5 could not yet be detected. The success of bacteriocin-negative S. bovis isolates was enhanced by the presence of another sensitive bacterium (Clostridium sticklandii SR). PCR based on repetitive DNA sequences indicated that S. bovis HC5 was not simply transferring bacteriocin genes to S. bovis JB1. When the two S. bovis strains were coinoculated into minimal medium, bacteriocin-negative isolates predominated, and this effect could be explained by the longer lag time (0.5 vs. 1.5 h) of S. bovis HC5. If the glucose concentration of the minimal medium was increased from 2 to 7 mg mL(-1), the effect of lag time was diminished and bacteriocin-producing isolates once again dominated the coculture. When the competition was examined in continuous culture, it became apparent that batch culture inocula were never able to displace a strain that had already reached steady state, even if the inoculum was large. This result indicated that bacterial selection for substrate affinity was even more important than bacteriocin production. PMID- 17117977 TI - Phosphorus-limited growth dynamics in two Baltic Sea cyanobacteria, Nodularia sp. and Aphanizomenon sp. AB - Rates of carbon (C) specific growth and nitrogen (N2) fixation were monitored in cultures of Baltic Sea Nodularia and Aphanizomenon exposed to gradual limitation by inorganic phosphorus (P). Both cyanobacteria responded by decreased cellular P content followed by lowered rates of growth and N2 fixation. C-specific growth and cellular N content changed faster in Aphanizomenon both when inorganic P was lowered as well as during reintroduction of P. Aphanizomenon also showed a more rapid increase in N-specific N2 fixation associated with increased C-specific growth. When ambient concentrations of inorganic P declined, both cyanobacteria displayed higher rates of alkaline phosphatase (APase) activity. Lower substrate half-saturation constants (KM) and higher Vmax : KM ratio of the APase enzyme associated with Nodularia suggest a higher affinity for dissolved organic P (DOP) substrate than Aphanizomenon. Aphanizomenon, which appears more sensitive to changes in ambient dissolved inorganic P, may be adapted to environments with elevated concentrations of P or repeated intrusions of nutrient-rich water. Nodularia on the other hand, with its higher tolerance to increased P starvation may have an ecological advantage in stratified surface waters of the Baltic Sea during periods of low P availability. PMID- 17117978 TI - Marine bacterioplankton production of polysaccharidic and proteinaceous particles under different nutrient regimes. AB - The influence of inorganic nutrient concentrations on the ability of bacterioplankton to produce and degrade polysaccharidic transparent exopolymer particles (TEPs) and proteinaceous Coomassie-stained particles (CSPs) was investigated in an 11-day experiment. The dynamics of these particles were followed in prefiltered (1 microm) northern Adriatic seawater enclosures enriched either with 1 microM orthophosphate (main limiting nutrient in this area), 10 microM ammonium or both orthophosphate and ammonium. These enclosures were referenced to a nonenriched control. A high potential for bacterial TEP and CSP production was observed (10(4) - 10(5) L(-1) for particles larger than 4 microm). In conditions of high orthophosphate concentration (either orthophosphate enriched or both orthophosphate and ammonium enriched), lower abundances and surface areas of CSPs were obtained, whereas TEP dynamics were more affected by unbalanced enrichments where only orthophosphate or ammonium was added. The impact of unbalanced nutrient ratios on TEPs was indicated by their higher abundance but low capacity for Alcian blue absorption, implying a change in their structure. Inorganic nutrient availability was thus proven to affect the bacterial potential for producing and degrading bacterially derived TEPs and CSPs. PMID- 17117979 TI - Carbon source utilization by the marine Dendryphiella species D. arenaria and D. salina. AB - Carbon utilization by the marine Dendryphiella species, D. arenaria and D. salina, was investigated to detect differences in utilization and traits associated with their adaptation to the marine habitat. Fifty-four strains were isolated world-wide and tested for the utilization of various carbon sources using BIOLOG phenotype MicroArray (PM) and for the production of extracellular enzymes on solid culture media and on API ZYM assay strips. PM analysis showed that the fastest growth occurred on several monosaccharides and amino acids, 2 keto-d-gluconic acid, succinamide and turanose. Some polyols were poor carbon sources. However, the two species differed in their utilization rates of carbon sources, forming three major clusters: two separate clusters for D. arenaria and D. salina and a third cluster in which strains of the two species formed separate subclades that correlated with geographic origin. Several carbon sources were also found useful in differentiating the two speices. Dendryphiella salina did not utilize xylitol and quinic acid, whereas D. arenaria grew well on these substrates. The latter failed to grow on sorbitol and grew slowly on mannitol, both were good substrates for the former. There were also no qualitative differences between the extracellular enzymes produced, although laccase and peroxidase activities were confined only to some strains. The physiological similarities exhibited by the two species support the close relationship between D. arenaria and D. salina. PMID- 17117980 TI - Effects of nutrient availability and Ochromonas sp. predation on size and composition of a simplified aquatic bacterial community. AB - Predation and competition are two main factors that determine the size and composition of aquatic bacterial populations. Using a simplified bacterial community, composed of three strains characterized by different responses to predation, a short-term laboratory experiment was performed to evaluate adaptations and relative success in communities with experimentally controlled levels of predation and nutrient availability. A strain with a short generation time (Pseudomonas putida), one with high plasticity in cell morphology (Flectobacillus sp. GC5), and one that develops microcolonies (Pseudomonas sp. CM10), were selected. The voracious flagellate Ochromonas sp. was chosen as a predator. To describe adaptations against grazing and starvation, abundance, biomass and relative heterogeneity of bacteria were measured. On the whole, the strains in the predation-free cultures exhibited unicellular growth, and P. putida represented the largest group. The presence of Ochromonas strongly reduced bacterial abundance, but not always the total biomass. The activity of grazers changed the morphological composition of the bacterial communities. Under grazing pressure the relative composition of the community depended on the substrate availability. In the presence of predators, P. putida abundance declined in both high and low nutrient treatments, and Pseudomonas CM10 developed colonies. Flectobacillus was only numerically codominant in the nutrient-rich environments. PMID- 17117981 TI - Effect of inoculum pretreatment on survival, activity and catabolic gene expression of Sphingobium yanoikuyae B1 in an aged polycyclic aromatic hydrocarbon-contaminated soil. AB - The survival and effectiveness of a bioaugmentation strain in its target environment depend not only on physicochemical parameters in the soil but also on the physiological state of the inoculated organism. This study examined the effect of variations in inoculum pretreatment on the survival, metabolic activity (measured as rRNA content) and polycyclic aromatic hydrocarbon (PAH)-catabolic gene expression of Sphingobium yanoikuyae B1 in an aged PAH-contaminated soil. RNA denaturing gradient gel electrophoresis analysis showed stable colonization of PAH-contaminated soil by S. yanoikuyae B1 after four pretreatments (growth in complex or minimal medium, starvation, or acclimation to phenanthrene). By contrast, extractable CFUs decreased with time for all four treatments, and significantly faster for Luria Bertani-grown inocula, suggesting that these cells adhered strongly to soil particles while remaining metabolically active. Pretreatment of the inoculum had a dramatic effect on the expression of genes specific to the PAH-degradation pathway. The highest levels of bphC and xylE expression were seen for inocula that had been precultivated on complex medium, and degradation of PAHs was significantly enhanced in soils treated with these inocula. The results suggest that using complex media instead of minimal media for cultivating bioaugmentation inocula may improve the subsequent efficiency of contaminant biodegradation in the soil. PMID- 17117982 TI - Synergy of two thermophiles enables decomposition of poly-epsilon-caprolactone under composting conditions. AB - The ultimate degradation (i.e. complete mineralization) of biodegradable polymers proceeds through hydrolysis to the production of degradation intermediates (primary degradation) that are then taken into the microbial cell and further degraded to CO2 and water. We first isolated thermophilic actinomycete (Streptomyces thermonitrificans PDS-1), which has the activity of ultimate degradability, from compost in which poly-epsilon-caprolactone (PCL) degraded vigorously. We next tried to investigate the detailed mechanisms of degradation of the PCL in compost by developing a new experimental method in which isolated microorganisms are used to inoculate sterilized compost raw materials containing PCL. It was confirmed that the ultimate degradation of PCL could not be achieved by the action of the strain PDS-1 alone, and that a supplementary microorganism (Bacillus licheniformis HA1) isolated from compost utilizes the degradation intermediates and also increases the activity of the other primary microorganism (PDS-1) by adjusting the pH. We could thus show experimental proof of synergy between two thermophiles in the ultimate degradation of a biodegradable polymer in compost. PMID- 17117983 TI - Persistence and mobility of a Clostridium botulinum spore population introduced to soil with spiked compost. AB - In a recent study it could be shown that compost samples can contain Clostridium botulinum. It was investigated if C. botulinum introduced with compost into botulinum-free soil can persist and be translocated within the soil. Compost was spiked with two C. botulinum type D spore concentrations (10(3) and 10(5) spores g(-1)) and the composts were spread on an experimental site. Over a period of 939 days, samples were taken from the upper (0-5 cm) and the lower (10-30 cm) soil horizons. Physical and chemical as well as microbiological variables were measured. Clostridium botulinum spores were quantified in a culture MPN-PCR assay. On day 757 the last positive sample was obtained in the plots with the lower spore concentration (10(3) g(-1)). The bacteria were never detected in the samples taken from the lower horizons of these plots. Clostridium botulinum persisted over the whole investigation period in the plots which were treated with compost spiked with 10(5) spores g(-1). The concentrations found were between 20 and 20,000 spores g(-1) soil. The bacteria were vertically translocated and could be found in the lower soil horizons (20-2000 spores g(-1) soil) starting 70 days after the compost was spread. PMID- 17117984 TI - Relationships between Fusarium population structure, soil nutrient status and disease incidence in field-grown asparagus. AB - Fusarium species cause important diseases in many crops. Lack of knowledge on how Fusarium species and strains interact with their environment hampers growth management strategies to control root diseases. A field experiment involving asparagus as host plant and three phosphorus fertilization levels was designed to examine the seasonal changes and ecological relationships between Fusarium populations and their soil and plant environments. Fusarium taxa were identified and assessed using PCR-denaturing gradient electrophoresis of the EF1-alpha gene. Resulting profiles were analyzed with respect to 17 ecological parameters measured during the three main asparagus phenological phases across a growing season. Multivariate statistical analysis showed that Fusarium population structure was strongly influenced by soil P level while seasonal variation was less important. A significant relationship between Fusarium population composition and Fusarium crown and root rot incidence was also found in September. Canonical analysis further revealed significant relationships between Fusarium population structure, and plant manganese and iron contents, soil dehydrogenase activity and soil calcium concentration. If higher Fusarium crown and root rot incidence is related to the Fusarium community structure, strategies to reduce the incidence in asparagus plantations may be found through manipulation of the soil fertility. PMID- 17117985 TI - Fungal endophytes in potato roots studied by traditional isolation and cultivation-independent DNA-based methods. AB - The composition and relative abundance of endophytic fungi in roots of field grown transgenic T4-lysozyme producing potatoes and the parental line were assessed by classical isolation from root segments and cultivation-independent techniques to test the hypothesis that endophytic fungi are affected by T4 lysozyme. Fungi were isolated from the majority of root segments of both lines and at least 63 morphological groups were obtained with Verticillium dahliae, Cylindrocarpon destructans, Colletotrichum coccodes and Plectosporium tabacinum as the most frequently isolated species. Dominant bands in the fungal fingerprints obtained by denaturing gradient gel electrophoresis analysis of 18S rRNA gene fragments amplified from total community DNA corresponded to the electrophoretic mobility of the 18S rRNA gene fragments of the three most abundant fungal isolates, V. dahliae, C. destructans and Col. coccodes, but not to P. tabacinum. The assignment of the bands to these isolates was confirmed for V. dahliae and Col. coccodes by sequencing of clones. Verticillium dahliae was the most abundant endophytic fungus in the roots of healthy potato plants. Differences in the relative abundance of endophytic fungi colonizing the roots of T4-lysozyme producing potatoes and the parental line could be detected by both methods. PMID- 17117986 TI - Spent media from cultures of environmental isolates of Escherichia coli can suppress the deficiency of biofilm formation under anoxic conditions of laboratory E. coli strains. AB - The prevailing lifestyle of bacteria is sessile and they attach to surfaces in structures known as biofilms. In Escherichia coli, as in many other bacteria, biofilms are formed at the air-liquid interface, suggesting that oxygen has a critical role in the biofilm formation process. It has been reported that anaerobically growing E. coli laboratory strains are unable to form biofilms even after 96 h of incubation on Luria Bertani (LB) medium. After analyzing 22,000 transposon-induced and 26,000 chemically-induced mutants we failed to isolate an E. coli laboratory strain with the ability to form biofilm under anaerobic growth conditions. Notably, seven strains from a collection of E. coli isolated from different hosts and the environment had the ability to form biofilm in the absence of oxygen. Interestingly, spent medium from cultures of one strain, Souza298, can promote biofilm formation of E. coli laboratory strains growing under anaerobic conditions. Our results led us to propose that laboratory E. coli strains do not release (or synthesize) a molecule needed for biofilm formation under anoxic conditions but that they bear all the required machinery needed for this process. PMID- 17117987 TI - Relevance of biofilm bacteria in modulating the larval metamorphosis of Balanus amphitrite. AB - Natural microbial communities found on different substrata exposed to the marine environment, including barnacle shell surfaces, are reported to have varying influences on the settlement and metamorphosis of competent cypris larvae. Experiments were carried out to compare the influence of settlement-inducing compounds from the bacteria isolated from the shell surface of Balanus amphitrite on its larval metamorphosis. The effect of multispecies bacterial film was also assessed. The production of different molecules by the bacteria was influenced by the nutrient media under which they were grown. It was observed that the promotory multispecies bacterial film turned to inhibition mode in the presence of the adult extract of the barnacle, indicating that bacteria-adult extract interactions alter the synthesis of different compounds produced by bacteria. The studies also show that the waterborne and the surface-associated cues from the bacteria function differentially in mediating larval metamorphosis. Understanding the complexities involved in such interactions and identification of the factors governing them would be a step forward. PMID- 17117988 TI - Evidence for the involvement of betaproteobacterial Thiobacilli in the nitrate dependent oxidation of iron sulfide minerals. AB - The Thiobacilli are an important group of autotrophic bacteria occurring in nature linking the biogeochemical cycles of sulfur and nitrogen. Betaproteobacterial Thiobacilli are very likely candidates for mediating the process of nitrate-dependent anoxic iron sulfide mineral oxidation in freshwater wetlands. A Thiobacillus denitrificans-like bacterium was present in an enrichment on thiosulfate and nitrate, derived from an iron-sulfide- and nitrate rich freshwater environment. Preliminary FISH analysis showed that the 16S rRNA gene-based bacterial probe mix showed great variation in intensity under different culture conditions. Furthermore, the widely applied 23S rRNA gene-based probe set BET42a/GAM42a incorrectly identified the T. denitrificans-like bacterium as a member of the Gammaproteobacteria. To circumvent these problems, the 23S rRNA genes of two T. denitrificans strains were partially sequenced and a new 23S rRNA gene-based probe (Betthio 1001) specific for betaproteobacterial Thiobacilli was designed. Use of this new probe Betthio 1001, combined with field measurements, indicates the involvement of Thiobacilli in the process of nitrate dependent iron sulfide mineral oxidation. PMID- 17117989 TI - Vertical distribution of nitrite reductase genes (nir S) in continental margin sediments of the Gulf of Mexico. AB - Marine sediments account for up to 66% of the loss of nitrogen load to coastal areas. Sedimentary denitrification is the main sink for fixed nitrogen in the global nitrogen budget, and thus it is important to understand the structure and composition of denitrifying communities. To understand the structure and composition of denitrifying communities, the diversity of nitrite reductase (nirS) genes from sediments along the Gulf of Mexico was examined using a PCR based cloning approach. Sediments were collected at three different depths (0 0.5, 4-5 and 19-21 cm). Geochemical analysis revealed decreasing nitrate and oxygen concentrations with increasing sediment depth. This trend coincided with the decrease in diversity of denitrifying bacteria. LIBSHUFF analysis indicated that the clone library in the shallowest sediment (depth, 0-0.5 cm) was significantly different from that in the deepest sediment (depth, 19-21 cm), and that the deeper sediments (depths of 4-5 and 19-21 cm) were significantly similar. Community structural shifts were evident between the shallowest (oxic zone) and deepest (anoxic zone) sediments. Community changes within the deepest sediments were more subtle, with the presence of different nirS clone sequences gradually becoming dominant or, alternatively, decreasing with depth. The changes in community structure at this depth are possibly driven by nutrient availability, with lower quality sources of carbon and energy leading to the disappearance of nirS sequences common in the top layer. The majority of recovered nirS sequences were phylogenetically divergent relative to known denitrifying bacteria in the database. PMID- 17117990 TI - Diversity estimates of microeukaryotes below the chemocline of the anoxic Mariager Fjord, Denmark. AB - Microbial communities of extreme environments have often been assumed to have low species richness. We analysed 18S rRNA gene signatures in a sample collected below the chemocline of the anoxic Mariager Fjord in Denmark, and from these data we computed novel parametric and standard nonparametric estimates of protistan phylotype richness. Our results indicate unexpectedly high richness in this environment: at the 99.5% phylotype definition, our most conservative estimate was 568 phylotypes (+/-114, standard error). Phylogenetic analyses revealed that the sequences collected cover the majority of described lineages in the eukaryotic domain. Out of 384 sequences analysed, 307 were identified as protistan targets, none of which was identical to known sequences. However, based on what is known about species that are phylogenetically related to the Mariager sequences, most of the latter seem to belong to strictly or facultative anaerobe organisms. We also found signatures that together with other environmental 18S rRNA gene sequences represent environmental clades of possibly high taxonomic levels (class to kingdom level). One of these clades, consisting exclusively of sequences from anoxic sampling sites, branches at the base of the eukaryotic evolutionary tree among the earliest eukaryotic lineages. Assuming eukaryotic evolution under oxygen-depleted conditions, these sequences may represent immediate descendants of early eukaryotic ancestors. PMID- 17117991 TI - Effect of ferritin overexpression in tobacco on the structure of bacterial and pseudomonad communities associated with the roots. AB - The genetic structures of total bacterial and pseudomonad communities were characterized in rhizosphere soil and rhizoplane+root tissues of tobacco wild type and a ferritin overexpressor transgenic line (P6) by a cultivation independent method using directly extracted DNA at the end of three consecutive plant cultures. The structure of total bacterial communities was characterized by automated ribosomal intergenic spacer analysis (A-RISA), and that of pseudomonad communities was characterized by PCR-restriction fragment length polymorphism (PCR-RFLP) from DNA amplified with specific primers. The structure of total bacterial communities was significantly modified in the rhizosphere soil by the overaccumulation of iron in the tobacco transgenic P6 line at the first culture, to a lesser extent at the second culture, and not at all at the third culture. No significant difference was recorded between the total communities associated with the roots (rhizoplane+root tissues) of the two plant genotypes in any of the cultures. In contrast, the difference in pseudomonad structure between the two plant genotypes increased with successive culture at the root level, but was not detected at a significant level in the rhizosphere soil. The impact of iron overaccumulation by the tobacco transgenic P6 line on pseudomonads supports previous findings on the importance of iron competition among fluorescent pseudomonads. PMID- 17117992 TI - Compaction of forest soil by logging machinery favours occurrence of prokaryotes. AB - Soil compaction caused by passage of logging machinery reduces the soil air capacity. Changed abiotic factors might induce a change in the soil microbial community and favour organisms capable of tolerating anoxic conditions. The goals of this study were to resolve differences between soil microbial communities obtained from wheel-tracks (i.e. compacted) and their adjacent undisturbed sites, and to evaluate differences in potential anaerobic microbial activities of these contrasting soils. Soil samples obtained from compacted soil had a greater bulk density and a higher pH than uncompacted soil. Analyses of phospholipid fatty acids demonstrated that the eukaryotic/prokaryotic ratio in compacted soils was lower than that of uncompacted soils, suggesting that fungi were not favoured by the in situ conditions produced by compaction. Indeed, most-probable-number (MPN) estimates of nitrous oxide-producing denitrifiers, acetate- and lactate-utilizing iron and sulfate reducers, and methanogens were higher in compacted than in uncompacted soils obtained from one site that had large differences in bulk density. Compacted soils from this site yielded higher iron-reducing, sulfate reducing and methanogenic potentials than did uncompacted soils. MPN estimates of H2-utilizing acetogens in compacted and uncompacted soils were similar. These results indicate that compaction of forest soil alters the structure and function of the soil microbial community and favours occurrence of prokaryotes. PMID- 17117993 TI - PCR DGGE and RT-PCR DGGE show diversity and short-term temporal stability in the Clostridium coccoides-Eubacterium rectale group in the human intestinal microbiota. AB - As the Clostridium coccoides-Eubacterium rectale (Erec; clostridial phylogenetic cluster XIVa) group is one of the major groups of the human intestinal microbiota, DNA- and RNA-based population analysis techniques (denaturing gradient gel electrophoresis; DGGE) were developed and applied to assess the diversity and temporal stability (6 months-2 years) of this faecal clostridial microbiota in 12 healthy adults. The stability of the Erec group was compared with the stability of the predominant bacterial microbiota, which was also assessed with PCR-DGGE. In addition, the Erec group was quantified with a hybridization-based method. According to our results, the Erec group was diverse in each subject, but interindividual uniqueness was not as clear as that of the predominant bacteria. The Erec group was found to be temporally as stable as the predominant bacteria. Over 200 clones obtained from two samples proved the developed method to be specific. However, the amount of bacteria belonging to the Erec group was not related to the diversity of that same bacterial group. In conclusion, the newly developed DGGE method proved to be a valuable and specific tool for the direct assessment of the stability of the Erec group, demonstrating diversity in addition to short-term stability in most of the subjects studied. PMID- 17117994 TI - 2,4-D impact on bacterial communities, and the activity and genetic potential of 2,4-D degrading communities in soil. AB - The key role of telluric microorganisms in pesticide degradation is well recognized but the possible relationships between the biodiversity of soil microbial communities and their functions still remain poorly documented. If microorganisms influence the fate of pesticides, pesticide application may reciprocally affect soil microorganisms. The objective of our work was to estimate the impact of 2,4-D application on the genetic structure of bacterial communities and the 2,4-D-degrading genetic potential in relation to 2,4-D mineralization. Experiments combined isotope measurements with molecular analyses. The impact of 2,4-D on soil bacterial populations was followed with ribosomal intergenic spacer analysis. The 2,4-D degrading genetic potential was estimated by real-time PCR targeted on tfdA sequences coding an enzyme specifically involved in 2,4-D mineralization. The genetic structure of bacterial communities was significantly modified in response to 2,4-D application, but only during the intense phase of 2,4-D biodegradation. This effect disappeared 7 days after the treatment. The 2,4-D degrading genetic potential increased rapidly following 2,4-D application. There was a concomitant increase between the tfdA copy number and the 14C microbial biomass. The maximum of tfdA sequences corresponded to the maximum rate of 2,4-D mineralization. In this soil, 2,4-D degrading microbial communities seem preferentially to use the tfd pathway to degrade 2,4-D. PMID- 17117995 TI - Microbial composition of chlorinated ethene-degrading cultures dominated by Dehalococcoides. AB - The community composition of microbial cultures degrading tetrachloroethene (PCE), trichloroethene (TCE), cis-1,2-dichloroethene (cDCE) and vinyl chloride (VC) to ethene was studied. A combination of PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and 16S rRNA gene sequence analysis revealed that all cultures contained Dehalococcoides populations, but that the populations of other organisms varied widely. Based on the sequences of cloned 16S rRNA genes, real time PCR methods were developed for several of these phylotypes affiliated with the putative dechlorinators Sulfurospirillum and Geobacter, the putative methanogens Methanomethylovorans, Methanomicrobiales, Methanosaeta and Methanosarcina, the putative acetogens Acetobacterium, Spirochaetes, and Sporomusa, and the putative fermenters Bacteroidetes, Syntrophus, and Syntrophobacter. These novel quantitative PCR methods were then used to estimate relative abundances of each phylotype in several individual cultures maintained on each chlorinated ethene. Dehalococcoides populations were the dominant phylotypes assayed in most KB-1 cultures, agreeing with the DGGE and cloning results. A Geobacter phylotype was also strongly represented in most PCE and TCE cultures, but not in cDCE or VC cultures, suggesting a possible role for this organism as a PCE-to-cDCE dechlorinator. The Sulfurospirillum phylotype was estimated to comprise a minor fraction of 16S rRNA gene copies and did not appear to have an important role in dechlorination. PMID- 17117996 TI - Molecular diversity studies of bacterial communities of oil polluted microbial mats from the Etang de Berre (France). AB - The biodiversity of microbial mats inhabiting the oil-contaminated lagoon Etang de Berre was determined by molecular approaches. The fingerprint of denaturing gradient gel electrophoresis (DGGE) and automatic ribosomal intergenic spacer analysis (ARISA) of mats exposed to different pollution levels showed specific microbial communities for each site but similar diversity richness. Species composition of the mats were compared by constructing 16S rRNA libraries. Amplified rDNA restriction analysis (ARDRA) of clone libraries confirmed their similar level of diversity richness. Phylogenetic analysis of the 16S rRNA sequences showed that the classes gamma and alpha of Proteobacteria were abundantly present in both sites whereas phylotypes related to the delta Proteobacteria and to the uncultured WS3 group were mainly found in the site with the highest pollution. Identification of the species involved in oil degradation by combining culture-based approaches and DGGE, showed that enrichment cultures were constituted by members of the Rhodobacterales and species related to Rhodococcus, Sphingomonas, Xanthomonas and Microbacterium, all of them known for their ability to degrade hydrocarbons. Our findings suggest that oil pollution has not affected the biodiversity richness of the mats. However, the populations involved in hydrocarbon degradation represent a minor fraction of the mat communities in the Etang de Berre. PMID- 17117997 TI - A longitudinal study of infant faecal microbiota during weaning. AB - For infants, the introduction of food other than breast milk is a high risk period due to diarrheal diseases, and may be corroborated with a shift in the faecal microbiota. This longitudinal study was the first undertaken to understand the effect of the supplementation on the infant's faecal microbiota and particularly the bifidobacteria. Eleven infants were enrolled. Their faecal microbiota were analysed using temporal temperature gradient gel electrophoresis (TTGE) with bacterial and bifidobacterial primers. In parallel, bifidobacterial counts were followed using competitive PCR. Three periods were distinguished: exclusive breastfeeding (Bf period), weaning (i.e. formula-milk addition, W period) and postweaning (i.e. breastfeeding cessation, Pw period). The bifidobacterial counts were not modified, reaching 10.5 (Log10 cells g(-1) wet weight). In the TTGE profiles, the main identified bands corresponded to Escherichia coli, Ruminococcus sp. and Bifidobacterium sp., more precisely Bifidobacterium longum, Bifidobacterium infantis and Bifidobacterium breve. For both TTGE profiles, the analysis of the distance suggested a maturation of the faecal microbiota but no correlation could be established with the diet. Despite a high interindividual variability, composition of the faecal microbiota appeared more homogenous after weaning and this point may be correlated with the cessation of breastfeeding. PMID- 17117998 TI - Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen. AB - Traditional methods for enumerating and identifying microbial populations within the rumen can be time consuming and cumbersome. Methods that involve culturing and microscopy can also be inconclusive, particularly when studying anaerobic rumen fungi. A real-time PCR SYBR Green assay, using PCR primers to target total rumen fungi and the cellulolytic bacteria Ruminococcus flavefaciens and Fibrobacter succinogenes, is described, including design and validation. The DNA and crude protein contents with respect to the fungal biomass of both polycentric and monocentric fungal isolates were investigated across the fungal growth stages to aid in standard curve generation. The primer sets used were found to be target specific with no detectable cross-reactivity. Subsequently, the real-time PCR assay was employed in a study to detect these populations within cattle rumen. The anaerobic fungal target was observed to increase 3.6-fold from 0 to 12 h after feeding. The results also indicated a 5.4-fold increase in F. succinogenes target between 0 and 12 h after feeding, whereas R. flavefaciens was observed to maintain more or less consistent levels. This is the first report of a real-time PCR assay to estimate the rumen anaerobic fungal population. PMID- 17117999 TI - Introduction of a boost of Legionella pneumophila into a stagnant-water model by heat treatment. AB - An environmentally representative stagnant-water model was developed to monitor the growth dynamics of Legionella pneumophila. This model was evaluated for three distinct water treatments: untreated tap water, heat-treated tap water, and heat treated tap water supplemented with Pseudomonas putida, a known biofilm-forming bacterium. Bringing heat-treated tap water after subsequent cooling into contact with a densely formed untreated biofilm was found to promote the number of L. pneumophila by 4 log units within the biofilm, while the use of untreated water only sustained the L. pneumophila levels. Subsequent colonization of the water phase by L. pneumophila was noticed in the heat-treated stagnant-water models, with concentrations as high as 1 x 10(10) mip gene copies L(-1) stagnant water. Denaturing gradient gel electrophoresis in combination with clustering analysis of the prokaryotic community in the water phase and in the biofilm phase suggests that the different water treatments induced different communities. Moreover, boosts of L. pneumophila arising from heat treatment of water were accompanied by shifts to a more diverse eukaryotic community. Stimulated growth of L. pneumophila after heating of the water may explain the rapid recolonization of L. pneumophila in water systems. These results highlight the need for additional or alternative measures to heat treatment of water in order to prevent or abate potential outbreaks of L. pneumophila. PMID- 17118000 TI - Exploitation of immunofluorescence for the quantification and characterization of small numbers of Pasteuria endospores. AB - The Pasteuria group of endospore-forming bacteria has been studied as a biocontrol agent of plant-parasitic nematodes. Techniques have been developed for its detection and quantification in soil samples, and these mainly focus on observations of endospore attachment to nematodes. Characterization of Pasteuria populations has recently been performed with DNA-based techniques, which usually require the extraction of large numbers of spores. We describe a simple immunological method for the quantification and characterization of Pasteuria populations. Bayesian statistics were used to determine an extraction efficiency of 43% and a threshold of detection of 210 endospores g(-1) sand. This provided a robust means of estimating numbers of endospores in small-volume samples from a natural system. Based on visual assessment of endospore fluorescence, a quantitative method was developed to characterize endospore populations, which were shown to vary according to their host. PMID- 17118001 TI - Beyond macronutrients: element variability and multielement stoichiometry in freshwater invertebrates. AB - We contrasted concentrations of macronutrients (C, N and P), essential (As, Cu, Zn and Se) and non-essential metals (Pb, Hg and Cd) in invertebrates across five lakes and June to October in one lake. We predicted that somatic concentrations of tightly regulated elements would be less variable than weakly and unregulated elements. Within each taxon, variation was lowest in macronutrients, intermediate in essential micronutrients, and highest in non-essential metals, which corresponded in rank to homeostatic regulation strength for the same elements calculated from the literature. Hence, homeostatic regulation may strongly influence variation in element concentrations of biota in situ. Of the individual elements, only taxonomic differences in C and N were consistent across lakes and over a season. Nevertheless, canonical discriminant analyses successfully discriminated among taxa based on taxonomic multielement composition. Thus, relative taxonomic differences in multielement composition appear more informative than absolute stoichiometric formulae when considering the role of inherently variable trace elements in ecological investigations. PMID- 17118002 TI - Community maturity, species saturation and the variant diversity-productivity relationships in grasslands. AB - Detailed knowledge of the relationship between plant diversity and productivity is critical for advancing our understanding of ecosystem functioning and for achieving success in habitat restoration efforts. However, effects and interactions of diversity, succession and biotic invasions on productivity remain elusive. We studied newly established communities in relation to preexisting homogeneous vegetation invaded by exotic plants in the northern Great Plains, USA, at four study sites for 3 years. We observed variant diversity-productivity relationships for the seeded communities (generally positive monotonic at three sites and non-monotonic at the other site) but no relationships for the resident community or the seeded and resident communities combined at all sites and all years. Community richness was enhanced by seeding additional species but productivity was not. The optimal diversity (as indicated by maximum productivity) changed among sites and as the community developed. The findings shed new light on ecosystem functioning of biodiversity under different conditions and have important implications for restoration. PMID- 17118003 TI - The role of exotic species in homogenizing the North American flora. AB - Exotic species have begun to homogenize the global biota, yet few data are available to assess the extent of this process or factors that constrain its advance at global or continental scales. We evaluate homogenization of vascular plants across America north of Mexico by comparing similarity in the complete native and exotic floras between states and provinces of the USA and Canada. Compared with native species, exotic plants are distributed haphazardly among areas but spread more widely, producing differentiation of floras among neighbouring areas but homogenization at greater distance. The number of exotic species is more closely associated with the size of the human population than with ecological conditions, as in the case of native species, and their distributions are less influenced by climate than those of native species. PMID- 17118004 TI - Above- and below-ground impacts of introduced predators in seabird-dominated island ecosystems. AB - Predators often exert multi-trophic cascading effects in terrestrial ecosystems. However, how such predation may indirectly impact interactions between above- and below-ground biota is poorly understood, despite the functional importance of these interactions. Comparison of rat-free and rat-invaded offshore islands in New Zealand revealed that predation of seabirds by introduced rats reduced forest soil fertility by disrupting sea-to-land nutrient transport by seabirds, and that fertility reduction in turn led to wide-ranging cascading effects on belowground organisms and the ecosystem processes they drive. Our data further suggest that some effects on the belowground food web were attributable to changes in aboveground plant nutrients and biomass, which were themselves related to reduced soil disturbance and fertility on invaded islands. These results demonstrate that, by disrupting across-ecosystem nutrient subsidies, predators can indirectly induce strong shifts in both above- and below-ground biota via multiple pathways, and in doing so, act as major ecosystem drivers. PMID- 17118005 TI - Energy, range dynamics and global species richness patterns: reconciling mid domain effects and environmental determinants of avian diversity. AB - Spatial patterns of species richness follow climatic and environmental variation, but could reflect random dynamics of species ranges (the mid-domain effect, MDE). Using data on the global distribution of birds, we compared predictions based on energy availability (actual evapotranspiration, AET, the best single correlate of avian richness) with those of range dynamics models. MDE operating within the global terrestrial area provides a poor prediction of richness variation, but if it operates separately within traditional biogeographic realms, it explains more global variation in richness than AET. The best predictions, however, are given by a model of global range dynamics modulated by AET, such that the probability of a range spreading into an area is proportional to its AET. This model also accurately predicts the latitudinal variation in species richness and variation of species richness both within and between realms, thus representing a compelling mechanism for the major trends in global biodiversity. PMID- 17118006 TI - Thermal range predicts bird population resilience to extreme high temperatures. AB - The identification of the characteristics of species that make them susceptible or resilient to climate change has been elusive because non-climatic influences may dominate short- and medium-term changes in population and distribution sizes. Here we studied the 2003 French heat wave, during which other confounding variables remained essentially unchanged, with a correlational approach. We tested the relationship between population resilience and thermal range by analysing the responses of 71 bird species to a 6-month heat wave. Species with small thermal ranges showed the sharpest decreases in population growth rate between 2003 and 2004 in locations with the highest temperature anomalies. Thermal range explained the resilience of birds to the heat wave independently of other potential predictors, although it correlated with nest location and broad habitat type used by species. The geographically deduced thermal range appears to be a reliable predictor of the resilience of these endothermic species to extreme temperatures. PMID- 17118007 TI - Sensitivity of the population growth rate to demographic variability within and between phases of the disturbance cycle. AB - For species in disturbance-prone ecosystems, vital rates (survival, growth and reproduction) often vary both between and within phases of the cycle of disturbance and recovery; some of this variation is imposed by the environment, but some may represent adaptation of the life history to disturbance. Anthropogenic changes may amplify or impede these patterns of variation, and may have positive or negative effects on population growth. Using stochastic population projection matrix models, we develop stochastic elasticities (proportional derivatives of the long-run population growth rate) to gauge the population effects of three types of change in demographic variability (changes in within- and between-disturbance-phase variability and phase-specific changes). Computing these elasticities for five species of disturbance-influenced perennial plants, we pinpoint demographic rates that may reveal adaptation to disturbance, and we demonstrate that species may differ in their responses to different types of changes in demographic variability driven by climate change. PMID- 17118008 TI - The rate of senescence in maternal performance increases with early-life fecundity in red deer. AB - Tradeoffs between reproduction and somatic maintenance are a frequently cited explanation for reproductive senescence in long-lived vertebrates. Between individual variation in quality makes such tradeoffs difficult to detect and evidence for their presence from wild populations remains scarce. Here, we examine the factors affecting rates of senescence in maternal breeding performance in a natural population of red deer (Cervus elaphus), using a mixed model framework to control for between-individual variance. Senescence began at 9 years of age in two maternal performance traits. In both traits, females that produced more offspring in early life had faster rates of senescence. This tradeoff is evident alongside significant effects of individual quality on late life breeding performance. These results present rare evidence in support of the disposable soma and antagonistic pleiotropy theories of senescence from a wild vertebrate population and highlight the utility of mixed models for testing theories of ageing. PMID- 17118009 TI - Florivory: the intersection of pollination and herbivory. AB - Plants interact with many visitors who consume a variety of plant tissues. While the consequences of herbivory to leaves and shoots are well known, the implications of florivory, the consumption of flowers prior to seed coat formation, have received less attention. Herbivory and florivory can yield different plant, population and community outcomes; thus, it is critical to distinguish between these two types of consumption. Here, we consider the ecological and evolutionary consequences of florivory. A growing number of studies recognize that florivory is common in natural systems and in some cases surpasses leaf herbivory in magnitude and impact. Florivores can affect male and female plant fitness via direct trophic effects and through altered pathways of species interactions. In particular, florivory can affect pollination and have consequences for plant mating and floral sexual system evolution. Plants are not defenceless against florivore damage. Concepts of resistance and tolerance can be applied to plant-florivore interactions. Moreover, extant theories of plant chemical defence, including optimal defence theory, growth rate hypothesis and growth differentiation-balance hypothesis, can be used to make testable predictions about when and how plants should defend flowers against florivores. The majority of the predictions remain untested, but they provide a theoretical foundation on which to base future experiments. The approaches to studying florivory that we outline may yield novel insights into floral and defence traits not illuminated by studies of pollination or herbivory alone. PMID- 17118010 TI - Activator of G-protein signaling in asymmetric cell divisions of the sea urchin embryo. AB - An asymmetric fourth cell division in the sea urchin embryo results in formation of daughter cells, macromeres and micromeres, with distinct sizes and fates. Several lines of functional evidence presented here, including pharmacological interference and dominant negative protein expression, indicate that heterotrimeric G protein Gi and its interaction partner, activator of G-protein signaling (AGS), are necessary for this asymmetric cell division. Inhibition of Gi signaling by pertussis toxin interferes with micromere formation and leads to defects in embryogenesis. AGS was isolated in a yeast two-hybrid screen with G alpha i as bait and was expressed in embryos localized to the cell cortex at the time of asymmetric divisions. Introduction of exogenous dominant-negative AGS protein, containing only G-protein regulatory (GPR) domains, selectively prevented the asymmetric division in normal micromere formation. These results support the growing evidence that AGS is a universal regulator of asymmetric cell divisions in embryos. PMID- 17118012 TI - Involvement of a Xenopus nuclear GTP-binding protein in optic primordia formation. AB - Using a subtracted Xenopus cDNA library based on the differential sensitivity of anterior and posterior genes to retinoic acid, we isolated a novel Xenopus nuclear GTP-binding protein (XGB). XGB is expressed prominently in the optic primordia at the tailbud stage. The N-terminal region of XGB contains a set of GTP-binding protein motifs, and the C-terminal region contains two putative nuclear localization signals and two coiled regions. A GFP-XGB fusion protein was expressed in the nucleus of NIH3T3 cells where it bound to subnuclear structures. Truncated C-terminal constructs of XGB containing both nuclear localization signal(s) and coiled region(s) suppressed eye formation, whereas neither the N terminal construct nor constructs with a mutated GTP-binding protein motif affected eye formation. Expression of Pax6 and Rx1 genes, which are crucial for eye development, was reduced in embryos overexpressing the C-terminal constructs of XGB. Suppression of Pax6 and Rx1 at earlier developmental stages as well as perturbation of eye formation at later stages was counteracted by co-expression of wild-type XGB. We conclude that XGB plays a role in the formation of optic primordia through activation of at least two eye field transcription factors. PMID- 17118011 TI - Increase in multidrug transport activity is associated with oocyte maturation in sea stars. AB - In this study, we report on the presence of efflux transporter activity before oocyte maturation in sea stars and its upregulation after maturation. This activity is similar to the multidrug resistance (MDR) activity mediated by ATP binding cassette (ABC) efflux transporters. In sea star oocytes the efflux activity, as measured by exclusion of calcein-am, increased two-fold 3 h post maturation. Experiments using specific and non-specific dyes and inhibitors demonstrated that the increase in transporter activity involves an ABCB protein, P-glycoprotein (P-gp), and an ABCC protein similar to the MDR-associated protein (MRP)-like transporters. Western blots using an antibody directed against mammalian P-gp recognized a 45 kDa protein in sea star oocytes that increased in abundance during maturation. An antibody directed against sea urchin ABCC proteins (MRP) recognized three proteins in immature oocytes and two in mature oocytes. Experiments using inhibitors suggest that translation and microtubule function are both required for post-maturation increases in transporter activity. Immunolabeling revealed translocation of stored ABCB proteins to the plasma cell membrane during maturation, and this translocation coincided with increased transport activity. These MDR transporters serve protective roles in oocytes and eggs, as demonstrated by sensitization of the oocytes to the maturation inhibitor, vinblastine, by MRP and PGP-specific transporter inhibitors. PMID- 17118013 TI - Phylogenetic correspondence of the body axes in bilaterians is revealed by the right-sided expression of Pitx genes in echinoderm larvae. AB - Chordates and echinoderms are two of the three major deuterostome phyla and show conspicuous left-right (LR) asymmetry. The establishment of LR asymmetry has been explored in vertebrates, but is largely unknown in echinoderms. Here, we report the expression pattern of genes that are orthologous to the chordate left-side specific gene Pitx, cloned from the sea urchin Hemicentrotus pulcherrimus (HpPitx) and the starfish Asterina pectinifera (ApPitx). HpPitx transcripts were first detected bilaterally in one cell of the ventrolateral primary mesenchyme cell aggregate of early prism larvae. New expression was detected asymmetrically in the right counterpart of a bilateral pair of mesodermal coelomic pouches and in the right ectoderm. In starfish bipinnaria larvae, the ApPitx signal was detected in the right coelomic pouch and in the right half of the ectoderm along the ciliary bands. These results suggest that the function of Pitx in establishing LR asymmetry was introduced in the last common ancestor of echinoderms and chordates. However, the right-side specific expression in echinoderm larvae is inverted compared to chordate embryos. This indicates that the LR axis is inversely represented between echinoderms and chordates, which supports the scenario that dorsoventral axis inversion was introduced into the chordate lineage by turning upside down. PMID- 17118014 TI - Expression and promoter analysis of Xenopus DMRT1 and functional characterization of the transactivation property of its protein. AB - The doublesex and mab-3-related transcription factor 1 (DMRT1) is involved in testis formation in a variety of vertebrates. In the teleost fish, Medaka, DMY/DMRT1Y on the Y chromosome, a duplicate of the autosomal DMRT1 gene, is characterized as a sex-determining gene. We report here the characterization of the Xenopus DMRT1 genes. Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that X. laevis DMRT1 was expressed throughout the embryo during early development and was restricted to the primordial gonads after embryogenesis. Whole-mount in situ hybridization analysis of the gene confirmed its specific expression in the primordial gonads. To study the transcriptional control of DMRT1 gene expression, we isolated the predicted promoter region of X. tropicalis DMRT1 using databases for this species. Analysis of transgenic tadpoles with a green fluorescence protein (GFP) reporter showed that approximately 3 kb of the 5'-flanking sequence of the DMRT1 gene was implicated in DMRT1 expression in the primordial gonads. We also showed that the C-terminal region of DMRT1 functioned as a transactivation domain in cultured cells, by a luciferase reporter assay using fusion proteins with the DNA-binding domain of GAL4. These findings suggest that DMRT1 functions as an activator of one or more genes involved in sex determination or gonadal differentiation. PMID- 17118015 TI - Development of calcium releasing activity induced by inositol trisphosphate and cyclic ADP-ribose during in vitro maturation of sea urchin oocytes. AB - During fertilization of sea urchin eggs, the cytoplasmic Ca(2+) concentration ([Ca(2+)](i)) transiently increases (Ca(2+) transient). Increased [Ca(2+)](i) results from a rapid release from intracellular stores, mediated by one or both of two signaling pathways; inositol 1,4,5-trisphosphate (IP(3)) and IP(3) receptor (IP(3)R) or cyclic GMP (cGMP), cyclic ADP-ribose (cADPR) and ryanodine receptor (RyR). During fertilization, cGMP and cADPR increase preceding the Ca(2+) transient, suggesting their contribution to this. If the RyR pathway contributed to the Ca(2+) transient, its Ca(2+) releasing activity would develop in parallel with that of the IP(3) system during maturation of oocytes. Sea urchin oocytes were cultivated in vitro and Ca(2+) transients induced by photolysis of caged IP(3) or caged cADPR were measured during maturation. Oocytes spontaneously began to maturate in seawater. More than 50% of oocytes underwent germinal vesicle breakdown within 25 h and the second meiosis within 35 h, but it took more than 24 h until they became functionally identical to in vivo-matured eggs. Both IP(3) and cADPR induced Ca(2+) transients comparable to those of in vivo-matured eggs later than 24 h from the second meiosis. However, cADPR induced a small Ca(2+) transient even before meiosis, whereas IP(3) and sperm almost did not. PMID- 17118016 TI - Identification and origin of the germline stem cells as revealed by the expression of nanos-related gene in planarians. AB - The planarian's remarkable regenerative ability is thought to be supported by the stem cells (neoblasts) found throughout its body. Here we report the identification of a subpopulation of neoblasts, which was revealed by the expression of the nanos-related gene of the planarian Dugesia japonica, termed Djnos. Djnos-expressing cells in the asexual planarian were distributed to the prospective ovary or testes forming region in the sexual planarian. During sexualization, Djnos-expressing cells produce germ cells, suggesting that in the asexual state these cells were kept as germline stem cells for the oogonia and spermatogonia. Interestingly, the germline stem cells were indistinguishable from the neoblasts by morphology and X-ray sensitivity and did not seem to contribute to the regeneration at all. Germline stem cells initially appear in the growing infant planarian, suggesting that germline stem cells are separated from somatic stem cells in the planarian. Thus, planarian neoblasts can be classified into two groups; somatic stem cells for regeneration and tissue renewal, and germline stem cells for production of germ cells during sexualization. However, Djnos-positive cells appeared in the newly formed trunk region from the head piece, suggesting that somatic stem cells can convert to germline stem cells. PMID- 17118017 TI - Improvement of stone comminution by slow delivery rate of shock waves in extracorporeal lithotripsy. AB - AIM: The aim of this study was to investigate the effect of delivery rate of shockwaves (SW) on stone comminution and treatment outcomes in patients with renal and ureteral stones. METHODS: Patients with radio-opaque stones in the upper urinary tract that were treated by extracorporeal shock wave lithotripsy (ESWL) were divided into two groups according to delivery rate (120 or 60 SW/min). The effective fragmentation after one ESWL session and treatment success at 3 months after ESWL was compared between the two groups. RESULTS: Of 134 patients (84 men and 50 women), 68 patients were treated at a fast rate and 66 were treated at a slow rate. Thirty and 38 patients in the fast rate group and 28 and 38 in the slow rate group had renal and ureteral stones, respectively. After one ESWL session, effective fragmentation was noted more often in the slow group (65.2%) than the fast group (47.1%) (P = 0.035), particularly for smaller stones (stone area <100 mm(2)) (P = 0.005) and renal stones (p = 0.005). However, there was no significant difference in treatment success at 3 months after ESWL between the two groups. In univariate logistic regression analysis, slow SW rate and smaller stones were significant factors for effective fragmentation after one ESWL session. In multivariate analysis, slow SW rate and smaller stones were also independent factors. CONCLUSIONS: Slow SW rate contributed to better stone comminution than fast rate, particularly for small stones and renal stones. ESWL treatment at a slow SW rate is recommended to obtain efficient stone fragmentation. PMID- 17118018 TI - Laparoscopic management of urachal remnants in adulthood. AB - BACKGROUND: The aim of this study was to investigate the outcome of laparoscopic excision of urachal remnants (LUR), and to compare the outcome with that of the traditional open excision of urachal remnants (OUR). METHODS: Between February 2001 and December 2005, six patients with a mean age of 23.8 years who had a symptomatic urachal sinus underwent radical LUR. Using 12 mm and 5 mm ports, the caudal stump of the urachus was ligated with an absorbable clip and divided. The peritoneal and preperitoneal tissue between the medial umbilical ligaments was dissected free of the transversalis fascia. Dissection was carried out along the preperitoneal plane toward the umbilicus. The cephalic side of the lesion was ligated at the umbilicus with an endo-loop and divided. In addition, four patients who underwent a traditional OUR were included. Peri- and postoperative records were reviewed to assess morbidity, recovery, and outcome. RESULTS: The operative duration was not significantly shorter for the LUR group than the OUR group, but there was generally a reduction in blood loss (mean 16.5 vs 68.3 mL), an earlier resumption of eating (mean 1.3 vs 2.5 days), and a shorter hospital stay (mean 5.3 vs 10.5 days). There were no intraoperative complications in either the LUR or the OUR group. Mean follow up was 5 (range 4-12) months. There were no postoperative complications. CONCLUSIONS: The results suggest that LUR can be safely and satisfactorily performed in adulthood. PMID- 17118019 TI - Comparison of two-port and four-port irradiation in the bladder preservation therapy for locally invasive bladder cancer. AB - AIM: To ascertain therapeutic effectiveness and adverse reactions with radiochemotherapy for locally invasive bladder cancer, comparison was made between two-port and four-port irradiation with intravenous cisplatin administration. METHODS: In 86 patients who were diagnosed with locally invasive bladder cancer on the basis of imaging and biopsy findings, transurethral tumor resection was carried out initially to minimize tumor volume, and then radiochemotherapy was done. From 1985 to 1997, bilateral two-port irradiation was carried out, and after 1998, four-port irradiation was done. In regards to chemotherapy, cisplatin was administered intravenously. Therapeutic effects were assessed 1 month after the end of therapy. RESULTS: Fifty-five patients underwent two-port irradiation; complete response (CR) was achieved in 26 and partial response (PR) in 29 patients. Thirty-one patients underwent four-port irradiation; CR was achieved in 20 patients and PR in 11 patients. The cause specific 5-year survival rate did not differ significantly between two-port and four-port irradiation (74%vs 79%), but there was a significant difference of survival between CR and PR patients. In the two-port irradiation group, 5-year bladder preservation rate was 89% in CR patients and 52% in PR patients. In the four-port irradiation group, 5-year bladder preservation rate was 90% in CR patients and 46% in PR patients. Of the various adverse reactions, no significant differences in leukopenia, thrombocytopenia or bladder symptoms were observed between two-port and four-port irradiation, but the incidence and severity of loss of appetite, nausea and/or vomiting, diarrhea and dermatitis were significantly greater for two-port irradiation. CONCLUSION: Radiochemotherapy are considered to be modest effective in the bladder preservation therapy for locally invasive bladder cancer. The four-port irradiation shows less adverse reactions than two-port irradiation. PMID- 17118020 TI - Lower urinary tract dysfunction in type 1 familial amyloidotic polyneuropathy in Kumamoto, Japan. AB - OBJECTIVE: To evaluate lower urinary tract dysfunction of type 1 familial amyloidotic polyneuropathy (FAP) patients in Kumamoto, Japan. METHODS: Lower urinary tract symptoms were evaluated in FAP patients. Urodynamic studies were evaluated in FAP patients as compared to those in control subjects. The location and distribution of amyloid deposits were evaluated in the urinary bladder in an autopsy case. RESULTS: In lower urinary symptoms, 86%, 19% and 38% patients showed difficulty in urination, urinary frequency and urinary incontinence. In detrusor function during filling cystometry, 14% patients showed detrusor overactivity. Moreover, 43% patients showed low compliance bladder, 62% and 38% patients showed normal and reduced bladder sensation, respectively. First desire to void (FDV), strong desire to void (SDV) and post-voided residual urine (PVR) were increased in FAP patients as compared to those in control subjects. In the urethral pressure profilometry, 71%, 10% and 19% patients showed incompetent, normal functional and overactive urethral closure mechanism, respectively. Maximum urethral pressure (MUP), maximum urethral closure pressure (MUCP) and functional profile length (FPL) were decreased in FAP patients compared to those in control subjects. CONCLUSION: Autonomic, somatic nerve systems and bladder detrusor musculature might be impaired in lower urinary tract of type 1 FAP patients in Kumamoto, Japan. PMID- 17118021 TI - Serum amino acids as indicators of cerebrospinal neuronal activity in patients with micturition disorders. AB - OBJECTIVE: Our previous study showed that the spinal glycine level in rats was changed by spinal injury or bladder outlet obstruction, and this change was reflected by serum glycine levels. Therefore, we measured the serum glutamate and glycine levels in healthy volunteers and patients with cerebrospinal damage or benign prostatic hyperplasia (BPH) to confirm whether the change of serum amino acid levels was obtained from these patients as well as the animal experiment. METHODS: We measured the serum glutamate and glycine levels in 170 healthy controls, 57 patients with cerebrovascular disease (CVD), 68 patients with spinal cord injury (SCI), and 70 patients with BPH. Amino acid levels were compared between the controls and patients, according to gender, level of spinal injury and the type of bladder activity. RESULTS: In the healthy controls, glutamate levels were higher and glycine levels were lower in men than in women. On group comparison of each gender, there were no differences of glutamate levels. However, glycine levels were lower in male and female SCI patients and BPH patients than in controls. According to the level of spinal injury or the pattern of bladder activity and amino acid levels, there were no relationships among them. CONCLUSIONS: Serum glutamate and glycine levels were not related to the spinal injury level or bladder activity. However, serum glycine levels changed in patients with SCI or BPH patients, so it may be possible to use it as an indicator of spinal glycinergic neuronal activity. PMID- 17118022 TI - Clinical study of complicated urinary tract infection using 'The UTI Criteria (Draft Fourth Edition)': measurement methods for pyuria. AB - OBJECTIVE: To evaluate the influences the change of the measurement method of pyuria from conventional centrifuged sediment to microchamber uncentrifuged urine for the results of evaluation of antimicrobial agents in clinical study against complicated urinary tract infections. From the viewpoint of international harmonization of judgement criteria, the recent method for counting white blood cells (WBC) in urine has changed from using uncentrifuged urine to using a microchamber in all countries. METHODS: Targeted diseases were non-catheterized complicated urinary tract infection, and cefcapene pivoxil hydrochloride or levofloxacin were used as antimicrobial drug. Pyuria was examined using the counting chamber method, a quantitative method using uncentrifuged urine with a microchamber, and the sedimentation method. RESULTS: Overall clinical efficacy in early evaluation by the two methods in measuring pyuria was evaluated as different in eight patients (7.3%). It was rated excellent in 63 (52.9%), moderate in 32 patients (26.9%) and poor in 24 (20.2%) with an efficacy rate of 79.8% using the counting chamber method, and excellent in 68 (57.1%), moderate in 27 (22.7%) and poor in 24 (20.2%) with an efficacy rate of 79.8% using the conventional sedimentation method CONCLUSION: No significant difference was seen between the two methods of WBC count in urine. PMID- 17118023 TI - Antibiotic prophylaxis in radical prostatectomy: 1-day versus 4-day treatments. AB - OBJECTIVE: The standard protocol of antibiotic prophylaxis in radical prostatectomy remains to be established. We retrospectively compared the occurrence of perioperative infections following radical prostatectomy between two different protocols of antibiotic prophylaxis. METHODS: This study included 106 cases of radical retropubic prostatectomy managed on the clinical pathways. Two different protocols of antibiotic prophylaxis were used in otherwise identical pathways. Between January and December 2004, 50 patients received a second generation cephem, cefotiam, for 4 days, beginning 30 min before surgery (4-day group), whilst between December 2004 and July 2005, only two doses of cefotiam were given on the day of operation in 56 patients (1-day group). The incidence of surgical site infection (SSI) and remote infection (RI) was retrospectively investigated. RESULTS: Superficial incisional SSI occurred in one (1.8%) patient in the 1-day group, whereas no patient in the 4-day group developed SSI. No RI was observed in either the 1-day or 4-day group. Intravenous antibiotics were administered besides the pathway in a patient in the 1-day group because unexplained fever more than 38 degrees C continued postoperative day (POD) 2 through POD 4 without signs of SSI or RI. Excluding this case, postoperative more than 38 degrees C was rare and transient after POD 2. CONCLUSION: The incidence of SSI and RI was low and not significantly different between the 1-day and 4-day groups. Therefore, the 1-day protocol of prophylactic antibiotic treatment seems adequate for preventing perioperative infections in radical prostatectomy. PMID- 17118024 TI - Efficacy of primary hormonal therapy for patients with localized and locally advanced prostate cancer: a retrospective multicenter study. AB - AIM: A retrospective review of patients with localized and locally advanced prostate cancer was performed to evaluate the efficacy of primary hormonal therapy and predict long-term prognosis in these patients. METHODS: A total of 628 patients who were diagnosed with stage T1c to T3 prostate cancer were treated with primary hormonal therapy at participating institutions. The patients were classified based on pretreatment prostate-specific antigen (PSA) level, Gleason score, and time to nadir PSA level. Disease-specific and progression-free survival rates were investigated, and compared among the subgroups. RESULTS: The mean age of patients was 74.5 years, and median pretreatment PSA level was 14.0 ng/mL. A total of 399 patients (63.5%) were treated with combined androgen blockade (CAB), and 229 patients (36.5%) were treated with castration monotherapy. The disease-specific survival rate of all 628 patients was 89.1% at 8 years. The group that showed a good response to primary hormonal therapy (Group G, pretreatment PSA level < or =20 ng/mL, Gleason score < or =7, and time to nadir PSA < or =6 months) accounted for approximately one-third of the total number of T1c-T3 patients. Disease-specific and progression-free survival rates at 8 years in Group G were 98.9% and 82.0%, respectively. These rates increased to 100% and 87.3%, respectively, in patients receiving CAB treatment in Group G. CONCLUSIONS: The results indicate the usefulness of primary hormonal therapy, especially CAB treatment, for patients showing a good response to hormonal therapy in long-term control of localized and locally advanced prostate cancer. PMID- 17118025 TI - Predictors of seminal vesicle invasion before radical prostatectomy. AB - AIM: To predict whether or not seminal vesicle invasion is present before radical prostatectomy, the relationships between clinical parameters and seminal vesicle invasion were analyzed. METHODS: A review was conducted of 187 patients who had been clinically diagnosed with stages A(2), B(0), B(1), B(2) or C prostate cancer and who had undergone radical prostatectomy without neoadjuvant therapy. The parameters analyzed for potential predictors of seminal vesicle invasion before radical prostatectomy included age, clinical stage, serum prostate-specific antigen (PSA) level at biopsy, tumor differentiation of biopsy specimens and percentage of cancer positive cores by biopsy. For percentage of cancer positive cores by biopsy, 143 of 187 patients who underwent transrectal sextant biopsy or more than six transrectal ultrasound guided core biopsies were evaluated. These parameters were subjected to univariate and multivariate logistic regression analyses to identify predictors for seminal vesicle invasion. RESULTS: The median age was 66.8 years (range 51-77 years). Of 187 patients, 27 (14.4%) had seminal vesicle invasion confirmed pathologically. There were significant differences in all parameters except for age between patients with positive and negative seminal vesicle invasion on univariate analysis. Multivariate analysis revealed that serum PSA level, tumor differentiation of biopsy specimens and percentage of cancer positive cores were significant independent predictors of seminal vesicle invasion. CONCLUSIONS: The results showed serum PSA level, tumor differentiation of biopsy specimens and percentage of cancer positive cores by biopsy before radical prostatectomy may be useful predictors for seminal vesicle invasion. PMID- 17118026 TI - Comparison of intravesical prostatic protrusion, prostate volume and serum prostatic-specific antigen in the evaluation of bladder outlet obstruction. AB - AIM: The aims of this study were to define the relationship between intravesical prostatic protrusion (IPP), prostate-specific antigen (PSA) and prostate volume (PV) and to determine which one of them is the best predictor of bladder outlet obstruction (BOO) due to benign prostatic enlargement. METHODS: A prospective study of 114 male patients older than 50 years examined between November 2001 and 2002 was performed. They were evaluated with digital rectal examination, International Prostate Symptoms Score, PSA, uroflowmetry, postvoid residual urine measurement, IPP and PV using transabdominal ultrasound scan. Statistical analysis included scatter plot with Spearman's correlation coefficients and nominal logistic regression RESULTS: Prostate volume, IPP and PSA showed parallel correlation. Although all three indices had good correlation with BOO index, IPP was the best. The Spearman rho correlation coefficients were 0.314, 0.408 and 0.507 for PV, PSA and IPP, respectively. Using receiver-operator characteristic curves, the areas under the curve for PV, PSA and IPP were 0.637, 0.703 and 0.772, respectively. The positive predictive values of PV, PSA and IPP were 65%, 68% and 72%, respectively. Using a nominal regression model, IPP remained the most significant independent index to determine BOO. CONCLUSIONS: All three non invasive indices correlate with one another. The study showed that IPP is a better predictor for BOO than PSA or PV. PMID- 17118027 TI - Tissue elasticity imaging for diagnosis of prostate cancer: a preliminary report. AB - BACKGROUND: Elastography is a diagnostic imaging technique that evaluates the hardness of a lesion. It is expected to become a new diagnostic modality for prostate cancer. The aim of this study was to examine the usefulness of elastography in the diagnosis of prostate cancer. METHODS: A total of 29 patients with untreated, histologically proven prostate cancer were examined using an elastographic imaging technique. The patient was scanned in the dorsosacral position and the prostate was manually compressed with a transrectal ultrasonic probe. The echo signals from inside the tissue were measured before and after the tissue compression and an elastogram was generated by spatially differentiation of the displacement distribution. RESULTS: Elastography depicted the cancer lesion as a harder tissue than the surrounding normal prostatic tissue. Elastography successfully detected 93% (27 patients) of the untreated prostate cancer lesions. Detection of cancer lesions using elastography was significantly higher than by digital rectal examination (59%; 17 patients) and transrectal ultrasonography (55%; 16 patients). CONCLUSION: Elastography has great potential as a useful modality for diagnosis of prostate cancer. Differentiation between cancerous and normal tissues can be expected to become more accurate as a result of technical advances in the quantification of tissue hardness. PMID- 17118028 TI - Intrathecal glutamate promotes glycinergic neuronal activity and inhibits the micturition reflex in urethane-anesthetized rats. AB - OBJECTIVES: In order to clarify the role of glutamate in the micturition reflex and in glutamatergic and glycinergic neuronal activity, we examined the effects of intrathecal (IT) injection of glutamate or MK-801 (an N-methyl-D-aspartate receptor antagonist) on bladder activity and on the glutamate and glycine levels in the lumbosacral cord of female rats with or without acute lower thoracic spinal cord injury (SCI). METHODS: Under urethane anesthesia, isovolumetric cystometry was performed in rats with or without SCI before and after IT injection of glutamate or MK-801 at the lumbosacral cord level. The glutamate and glycine levels of the whole lumbosacral cord were measured after IT injection of glutamate or MK-801 in both groups. RESULTS: In intact rats, IT glutamate (100 microg) prolonged the interval between bladder contractions and decreased the amplitude of contractions. IT MK-801 (3-100 microg) also prolonged the interval between bladder contractions and decreased the amplitude in intact rats. In SCI rats, cystometry demonstrated the disappearance of bladder contractions, and the glycine level in the lumbosacral cord was elevated. In intact rats, IT glutamate (0.3-100 microg) increased the glycine level in the lumbosacral cord. On the other hand, IT MK-801 (3-100 microg) decreased both glutamate and glycine levels in intact and SCI rats. CONCLUSIONS: These results suggest that glutamatergic neurons have stimulatory projections to both glutamatergic and glycinergic neurons in the lumbosacral cord, and that glutamatergic neurons inhibit the micturition reflex by stimulating glycinergic neurons. PMID- 17118029 TI - Solitary, late metastatic recurrence of renal cell carcinoma: two extraordinary cases. AB - Late recurrence of renal cell carcinoma (RCC) has been well documented in the literature. We present two extraordinary cases of solitary, late metastatic recurrence of RCC. The first is a case of a solitary, adrenal metastasis excised 38 years after nephrectomy and the second is a case in which two solitary metastatic deposits were resected 14 and 26 years after excision of the primary tumor. In each of these patients the solitary metastases were initially believed to be primary tumors at other sites; however, on histological examination they were found to be metastatic RCC recurrences. In patients with a previous history of RCC presenting with apparently new solitary lesions, metastatic RCC must first be excluded. PMID- 17118030 TI - Carcinosarcoma of the ureter: report of a case with unusual histology and review of the literature. AB - Carcinosarcomas of the ureter are rare biphasic neoplasms, with distinct malignant epithelial and mesenchymal components. To the authors' knowledge there have been only 12 cases reported in the literature. Primary cartilaginous tumors of the ureter are also rare tumors. Thus far there has been a single case report of an extraskeletal mesenchymal chondrosarcoma of the ureter in the German literature. Herein a unique case is reported of ureteral carcinosarcoma in an 81 year-old woman presenting with gross hematuria in which the sarcomatous component had extraskeletal mesenchymal chondrosarcomatous differentiation. This is thought to be the first reported case of a carcinosarcoma with such a differentiation in the sarcomatous component. PMID- 17118031 TI - Primary adenocarcinoma of the rete testis with preceding diagnosis of pulmonary metastases. AB - We report a case of primary adenocarcinoma of the rete testis in a 55-year-old man with pulmonary metastases that were detected 11 months prior to the diagnosis of the primary lesion. Primary adenocarcinoma of the rete testis is an extremely rare malignant tumor with a poor outcome. The most common primary symptom is a scrotal mass, often accompanied by hydrocele and chronic epididymitis. The diagnosis is often delayed because of non-specific clinical presentation and symptoms. We cannot forget that rete testis is a possible primary site for a primary, unknown metastatic adenocarcinoma. PMID- 17118032 TI - Intratesticular borderline serous tumor. AB - Neoplasms resembling ovarian serous tumors rarely arise in the male genital organs. Mullerian histogenesis of this tumor has been suggested. We report a case of an intratesticular borderline serous tumor accidentally discovered during the course of a preoperative workup in a 72-year-old man admitted for left upper lobectomy for squamous cell carcinoma of the lung. PMID- 17118033 TI - Splenogonadal fusion: case report and review of published works. AB - Splenogonadal fusion is a rare congenital anomaly. We report herein a case of splenogonadal fusion associated with contra lateral testicular aplasia, and review the etiology, pathogenesis and management of this rare disease. As far as we know, this is the first reported case of splenogonadal fusion associated with testicular aplasia. PMID- 17118034 TI - Proximal-type epithelioid sarcoma of the perineum: calling attention of urologists. AB - A subcutaneous mass in the perineum of a middle-aged man was excised and pathologically diagnosed at first as 'undifferentiated carcinoma' of unknown origin, which recurred 2 years later without any metastasis. Further histological evaluation ultimately established a correct diagnosis of 'proximal-type epithelioid sarcoma', a variant of rare epithelioid sarcoma. This type of tumor may confuse pathologists because its histological characteristics resemble undifferentiated carcinoma or malignant rhabdoid tumor. Frequent immunoreactivity of CD34, in addition to expression of keratins, epithelial membrane antigen and vimentin, provides strong support for the diagnosis of this rare neoplasm. Urologists should be aware that this sarcoma commonly occurs in the genital regions. PMID- 17118036 TI - Chromosome arrangement and nuclear architecture but not centromeric sequences are conserved between Arabidopsis thaliana and Arabidopsis lyrata. AB - In contrast to the situation described for mammals and Drosophila, chromosome territory (CT) arrangement and somatic homologous pairing in interphase nuclei of Arabidopsis thaliana (n = 5) are predominantly random except for a more frequent association of the chromosomes bearing a homologous nucleolus organizer region. To find out whether this chromosome arrangement is also characteristic for other species of the genus Arabidopsis, we investigated Arabidopsis lyrata ssp. lyrata (n = 8), one of the closest relatives of A. thaliana. First, we determined the size of each chromosome and chromosome arm, the sequence type of centromeric repeats and their distribution between individual centromeres and the position of the 5S/45S rDNA arrays in A. lyrata. Then we demonstrated that CT arrangement, homologous pairing and sister chromatid alignment of distinct euchromatic and/or heterochromatic regions within A. lyrata interphase nuclei are similar to that in A. thaliana nuclei. Thus, the arrangement of interphase chromosomes appears to be conserved between both taxa that diverged about 5 million years ago. Since the chromosomes of A. lyrata resemble those of the presumed ancestral karyotype, a similar arrangement of interphase chromosomes is also to be expected for other closely related diploid species of the Brassicaceae family. PMID- 17118037 TI - Somali and Oromo refugee women: trauma and associated factors. AB - AIM: This paper reports a study identifying the demographic characteristics, self reported trauma and torture prevalence, and association of trauma experience and health and social problems among Somali and Oromo women refugees. BACKGROUND: Nearly all refugees have experienced losses, and many have suffered multiple traumatic experiences, including torture. Their vulnerability to isolation is exacerbated by poverty, grief, and lack of education, literacy, and skills in the language of the receiving country. METHOD: Using data from a cross-sectional population-based survey, conducted from July 1999 to September 2001, with 1134 Somali and Oromo refugees living in the United States of America, a sub-sample of female participants with clearly identified parenting status (n = 458) were analysed. Measures included demographics, history of trauma and torture, scales for physical, psychological, and social problems, and a post-traumatic stress symptom checklist. FINDINGS: Results indicated high overall trauma and torture exposure, and associated physical, social and psychological problems. Women with large families reported statistically significantly higher counts of reported trauma (mean 30, P < 0.001) and torture (mean 3, P < 0.001), and more associated problems (P < 0.001) than the other two groups. Women who reported higher levels of trauma and torture were also older (P < 0.001), had more family responsibilities, had less formal education (P < 0.001) and were less likely to speak English (P < 0.001). CONCLUSION: These findings suggest a need for nurses, and especially public health nurses who work with refugee and immigrant populations in the community, to develop a more comprehensive understanding of the range of refugee women's experiences and the continuum of needs post migration, particularly among older women with large family responsibilities. Nurses, with their holistic framework, are ideally suited to partner with refugee women to expand their health agenda beyond the biomedical model to promote healing and reconnection with families and communities. PMID- 17118038 TI - Women's perceptions of partner support and conflict in the development of postpartum depressive symptoms. AB - AIM: This paper reports a study examining the influence of maternal perceptions of conflict and relationship- and postpartum-specific support from the partner on the development of depressive symptoms in the first 8-weeks postpartum. BACKGROUND: Although the quality of a woman's relationship with her partner has been associated with risk for postpartum depression, few studies have examined which elements of partner-specific support and conflict are determinants of maternal mental health. METHODS: Self-administered measures of partner support (Social Provisions Checklist, Postpartum Partner Support Scale), partner conflict (Quality of Relationships Inventory), and depression (Edinburgh Postnatal Depression Scale) were mailed postnatally to a population-based sample of 396 mothers between April 2001 to January 2002. Differences in partner support and conflict at 1 and 4-weeks postpartum were examined between those women who met criteria for probable depression (Edinburgh Postnatal Depression Scale > 9) and those who did not meet criteria for probable depression at 8-weeks postpartum. FINDINGS: Women with depressive symptoms at 8-weeks postpartum had significantly lower perceptions of relationship-specific and postpartum-specific partner support and significantly higher levels of relationship conflict than women with no depressive symptoms. Multiple regression analyses revealed that three variables, perceived social integration, partner encouragement to obtain help when needed, and partner agreed with infant care, significantly explained the variance in Edinburgh Postnatal Depression Scale scores. CONCLUSIONS: Maternal perceptions of social integration and partner provision of problem-focused informational support and positive feedback are important in determining maternal mental health. Therefore, in the antenatal period healthcare professionals should teach couples appropriate ways to provide feedback and communicate expectations, especially those relating to infant care strategies. The importance of appraisal and emotional support should be highlighted in order to foster maternal feelings of acceptance and being cared for. Partners should also be encouraged to participate actively in household tasks and infant care activities to protect the mother from becoming overwhelmed. PMID- 17118039 TI - Organizational predicaments: ethical conditions for nursing research. AB - AIM: This paper highlights the sometimes problematic matter of access to the field in clinical ethnographic research by discussing a hypothetical scenario of an instance of change in the terms of access. BACKGROUND: A review of the methodological literature about research access revealed that there was little in the nursing literature about this issue, although anecdotally many nurse researchers reveal that access is not unproblematic and requires active maintenance to ensure smooth operation of a project. However, in returning to literature in sociology and anthropology, where gaining access for fieldwork is not considered so routine, we found much in the literature about dilemmas, predicaments and conflicts over access, confidentiality and anonymity that were not mentioned in nursing research about access to clinical areas. We returned also to the ethical and legal framework guiding access practices so as to explore the issues in fieldwork situations. DISCUSSION: In exploring the problems arising from maintaining confidentiality, safety of participants and researchers, it is clear that the much-vaunted insider status is both a boon and a burden. Intimate and inquisitive research using ethnographic techniques has the potential to unsettle organizations and workers through the very processes used to undertake the research. CONCLUSION: Ethical comportment is not enough to protect a study. A more overt and less naive approach to access for fieldwork is required to better inform future nurse researchers how to maintain access to the field and the requirement of constant negotiation and adjustment. PMID- 17118040 TI - Exploring painful experiences: impact of emotional narratives on members of a qualitative research team. AB - AIM: This paper reports a study of the impact of emotional narratives on the well being of members of a qualitative research team during the conduct of sensitive research. BACKGROUND: Qualitative data are frequently collected from participants using repeated in-depth interviews when exploring sensitive issues such as loss and grief. The research process can evoke highly emotional responses in the participant and others involved in the study. While consideration has been given to the impact of the research process on participants when a highly affective component is involved, relatively little attention has been given to research team members' experiences. METHOD: Through analysis of fieldwork records from a grounded theory study of the experiences of women who were carrying a baby with a foetal abnormality, we discuss the affective issues arising in conducting sensitive research. Data sources included two reflexive journals, written comments from two transcribers and the transcript of an interview with the research supervisor. FINDINGS: The core category of 'Connecting with the data' emerged, to which each substantive category relates. Three substantive categories -'bearing to watch,''bearing to listen' and 'bearing to support'- emerged as independent but inter-related aspects of the research process as experienced by the researcher, transcribers and supervisor. Methods of protecting the research team and the integrity of the study when the substantive issue is highly emotive are discussed. CONCLUSION: The emotional impact of research on participants is normally considered prior to the conduct of any sensitive research, and efforts are made to protect them. The potential for researchers, transcribers and supervisors to be harmed should also be carefully considered when planning a project with significant affective elements. PMID- 17118041 TI - Effectiveness of non-pharmacological interventions for fatigue in adults with multiple sclerosis, rheumatoid arthritis, or systemic lupus erythematosus: a systematic review. AB - AIM: This paper reports a systematic review of non-pharmacological interventions for fatigue in adults with three common autoimmune conditions. BACKGROUND: A considerable proportion of people with multiple sclerosis, rheumatoid arthritis, and systemic lupus erythematosus experience compromised quality of life due to fatigue. Recent reviews of pharmacotherapies for fatigue in these conditions remain inconclusive, and systematic evidence for effectiveness of non pharmacological interventions was unavailable. Our paper addresses this gap. METHODS: The literature search used the key words fatigue, energy, multiple sclerosis, rheumatoid arthritis and systemic lupus. It included 19 electronic databases and libraries, three evidence-based journals, two internet search engines, was dated 1987-2006, and limited to English. Non-pharmacological experimental studies about fatigue comprising more than five adults were included. Meta-analysis was not possible due to diverse interventions and outcome measures, therefore studies were analysed by types of interventions used to reduce fatigue. RESULTS: Of 653 hits, 162 papers were reviewed, and 36 met the inclusion criteria. Thirty-three primary studies reported 14 randomized controlled trials and 19 quasi-experimental designs. Most interventions were tested with people with multiple sclerosis. Exercise, behavioural, nutritional and physiological interventions were associated with statistically significant reductions in fatigue. Aerobic exercise was effective, appropriate and feasible for reducing fatigue among adults with chronic autoimmune conditions. Electromagnetic field devices showed promise. The diversity of interventions, designs, and using 24 different instruments to measure fatigue, limited comparisons. CONCLUSION: Low impact aerobic exercise gradually increasing in intensity, duration and frequency may be an effective strategy in reducing fatigue in some adults with chronic auto-immune conditions. However, fatigue is a variable and personal experience and a range of behavioural interventions may be required. Well-designed studies testing these promising strategies and consensus on outcome fatigue measures are needed. PMID- 17118042 TI - What over-the-counter preparations are pregnant women taking? A literature review. AB - AIM: This paper presents an overview of current literature relating to over-the counter and herbal preparations use, with a focus on pregnancy. BACKGROUND: Internationally, there has been a shift towards self-medication through over-the counter release of many prescription-category drugs and wider acceptance and use of herbal preparations. The general community perception that these preparations are safe may lead to inappropriate use, especially during pregnancy. METHOD: A range of databases was searched to identify papers addressing 'over-the-counter preparations/medications', 'non-prescription medications', 'herbal preparations' and 'pregnancy', including Medline, Cumulative Index to Nursing, Allied Health Literature, Proquest and Sciencedirect. Midwifery, nursing and scientific papers published from 2000 onwards were included, along with a small number of scientific papers published from 1995 onwards, and some key textbooks to clarify the definitions. FINDINGS: Pregnant women use a range of over-the-counter preparations, including analgesics, antihistamines, antacids and a variety of herbal preparations. However, in many cases, it is unclear whether their use is actually safe in the short- or long-term for mother or baby, owing to an inadequate evidence-base to support the use of these preparations during pregnancy. CONCLUSION: While there is extensive literature on the use of over-the counter preparations generally, less is available about their use in pregnancy. Further research is needed to explore the prevalence of use and effects of these preparations in pregnancy and neonatal outcomes, and to support midwives and nurses in harm minimization through the promotion of informed risk-avoidance behaviours. PMID- 17118043 TI - Impact of recurrent changes in the work environment on nurses' psychological well being and sickness absence. AB - AIM: This paper is a report of a study of how the occurrence and appraisal of recurrent changes in the work environment of hospital nurses affect psychological well-being (i.e. job satisfaction, eustress and distress) and absence through illness. BACKGROUND: Many researchers have demonstrated the impact of major organizational changes on employees' psychological well-being, but only a few have focused on the permanent consequences in work conditions. In a contemporary healthcare setting, an increased number of recurrent operational changes has become a normal characteristic of nurses' work environment. Specific work situations have frequently been associated with occupational stress, whereas employees' appraisal of recurrent changes as stressors and their relation to psychological well-being and health outcomes (i.e. sickness absence) have been dismissed. METHODS: A cross-sectional questionnaire survey was conducted in 2003 with 2094 Registered Nurses in 10 general hospitals. Logistic regressions were used to investigate the impact on psychological well-being and prospectively measured rates of sickness absence (frequency and duration). RESULTS: The occurrence of changes in the work environment (in the past 6 months) had had a negative impact on staff psychological well-being. Nurses who had been confronted with changes scored statistically significantly higher for distress. Changes appraised as threatening were negatively related to job satisfaction and eustress, and positively related to distress and sickness absence (frequency and duration). Changes appraised as challenging were positively related to job satisfaction and eustress but had no impact on distress and sickness absence. CONCLUSION: Future research should take into consideration the impact of the occurrence and appraisal of recurrent changes in the work environment of healthcare employees (i.e. Registered Nurses) on psychological well-being and sickness absence. This should also be considered by managers when dealing with these nursing workforce issues. PMID- 17118044 TI - Does supplementary prenatal nursing and home visitation reduce healthcare costs in the year after childbirth? AB - AIM: This paper reports the costs of a programme of supplementary prenatal care, including healthcare costs, in the year following childbirth. BACKGROUND: Publicly funded healthcare systems have provided pregnant women with adequate medical care, but access to resources to address their non-medical needs is still an issue. To improve women's access to pregnancy-related resources, a community based, prenatal programme involving consultations with a specialist nurse, or nurse plus a home visitor was evaluated. METHOD: A sample of 284 women who had participated in a randomized controlled trial of the prenatal care programme participated in this partial economic analysis. Women had been randomized to one of three trial arms: (1) standard care, (2) standard care plus consultations with a specialist prenatal care nurse, or (3) standard care plus nurse consultations and a home visitor. For the economic study, each woman was asked about her and her baby's use of healthcare services in the 12 months after the baby's birth. Health service utilization was multiplied by the unit cost of each service and summed to arrive at the total cost of services used. The study was undertaken in 2004. RESULTS: Supplementary prenatal care neither increased the use of health services nor resulted in savings in health spending. Compared with standard care, women in the two intervention groups made more use of family physicians and less use of paediatricians, but no significant differences in the overall costs of health care were noted. CONCLUSION: While supplementary prenatal care had no impact on costs, some benefits occurred for those at greatest risk of not accessing services. However, it would be premature to draw widespread recommendations for policy from the results of a single study. Further investment in prenatal care should continue to be accompanied by rigorous evaluation of its costs and the value that women place on the service provided. PMID- 17118045 TI - The clinical nursing competences and their complexity in Belgian general hospitals. AB - AIM: This paper reports a study whose aim was to chart clinical nursing competences and their complexity in Belgian general hospitals. BACKGROUND: Competence is an essential factor for assuring quality, safety and cost-effective health care. As clinical competence cannot be evaluated separately from the clinical context in which decisions are made, competence is defined as functional adequacy and the capacity to integrate knowledge and skills with attitudes and values into the specific contexts of practice. METHOD: The study took place in 2003 and focused on basic care, specialized care and elder care. Head nurses of 176 departments processed 521 patient situations. Experts selected 50 situations per area. In a Delphi procedure 100 experts described the nursing competences required in each situation necessary to provide quality acceptable care. The experts determined the global complexity of each of these competences as well as the complexity of the cognitive, psychomotor and affective aspects. FINDINGS: The global complexity of the various care areas was similar. Cognitive and affective aspects of competences scored higher than psychomotor aspects. Simple and average situations did not show any significant difference with regard to complexity and were less complex than difficult situations, which obtained the highest complexity score. The complexity of emergency situations did not differ from that of difficult situations. CONCLUSION: Emergency and difficult situations require more competences than offered by basic nursing education. The continuous presence of care providers with additional education is necessary for each care area to respond adequately to emergency and difficult situations. Nurses with additional academic qualifications need to be available for consultation. There are only a few possibilities of employing lower qualified staff in direct care, with the exception of support with regard to administrative tasks. Cognitive and affective aspects of the competences require particular attention during the education programme. PMID- 17118046 TI - Disentangling the effects of psychological and physical work demands on sleep, recovery and maladaptive chronic stress outcomes within a large sample of Australian nurses. AB - AIM: This paper reports a study to determine if different types of work strain experienced by nurses, particularly those of an essentially psychological nature, such as emotional demand, mental effort and problems with peers and/or supervisors, have a differential impact on sleep quality and overall recovery from work strain, compared with physical work strains, and lead to higher maladaptive chronic fatigue outcomes. BACKGROUND: Various studies have shown that the dominant work-demand strain associated with nursing work can vary between different areas of nursing. For example, whereas emotional strain is reported to be the principal strain associated with work in areas such as oncology, haematology and renal units, medical and surgical unit nurses report work pace and staffing issues as the dominant work strain. Purely physical strain seems to be less commonly reported as a concern. METHOD: A large sample (n = 760) of Australian nurses working in a large metropolitan hospital completed questionnaires on their work demands, sleep quality, fatigue, and recovery between shifts in January 2004. FINDINGS: A high work pace exacerbates the psychological rather than the physical strain demands of nursing. Psychological strain affects sleep quality and impairs recovery from overall work strain between shifts. This combination is highly predictive of serious maladaptive stress/fatigue outcomes among nurses. CONCLUSION: Coping with psychological stressors adequately is an important requirement for nurses in order to avoid adverse health effects and maintain a long-term career in nursing. Appropriate training of undergraduate nursing students in managing the stresses they are likely to encounter would seem to be an essential requirement for the 21st century. Such training might constitute an important long-term component in overcoming the chronic nurse shortages evident in many countries. PMID- 17118050 TI - Two terms--one meaning: the conundrum of contemporary nomenclature in autism . PMID- 17118051 TI - Learning to observe in context: child and adolescent inpatient mental health assessment. AB - On child and adolescent inpatient psychiatric units, nursing staff have 24-hour exposure to patients, an excellent vantage point for observing behavior in a variety of situations and interpersonal contexts. How staff members respond to milieu behaviors depends in part on their judgment of what prompted a youngster's response and his/her ability to process the event and control the attendant emotions. To assess such aspects of a child or adolescent's presentation requires that staff appraise not just the presenting behavior but the patient's information-processing and affect-regulation abilities. One way this developmental knowledge can be embedded in practice is through assessment guides that contain observation cues for key issues related to how children/adolescents think, feel, and behave. This article sets down the rationale for the content and organization of such inpatient nursing assessment guides and suggestions for their dissemination to the unit staff. PMID- 17118052 TI - Following the affect: learning to observe emotional regulation. AB - On inpatient psychiatric units, staff deal with children and adolescents whose affect escalates quickly and intensely. These same children experience strong emotions that they can neither understand nor explain. To intervene effectively, inpatient staff must understand the regulation issues underneath the escalated behaviors. Emotion Regulation theory and the developmental line of emotional understanding are useful concepts in assessing and intervening with these children and adolescents. Presented here are criteria to guide inpatient staff's assessment of children and adolescents with emotion regulation difficulties. The assessment cues are based in concepts of Emotion Regulation and emotional understanding and are accompanied by suggested intervention strategies. PMID- 17118053 TI - Learning to observe cognition, mastery, and control. AB - On inpatient psychiatric units, intervening with children and adolescents' behaviors demands an understanding of how they take in and process information. The third paper focuses on assessment cues that will help inpatient staff decipher patients' deficits and strengths in information processing, self efficacy, and inhibitory control. Each assessment area is explained in the context of the supporting cognitive, neurobiologic, and social-learning theory. Inpatient interventions consistent with the assessment are explained and illustrated with clinical applications. PMID- 17118054 TI - Learning to observe relationships and coping. AB - Milieu relationships provide the critical background presence to staff's attempts to motivate, regulate, and teach patients how to cope with stress. Forging a connection with hospitalized children and adolescents demands attention to how they respond to adults and engage with staff around milieu expectations. Assessment guides that deal with these issues are presented. Important aspects of children's relatedness are presented in the context of their working models of adults and the influence of these representations on their response to staff. Coping skills are explained with particular emphasis on behavioral coping strategies. Tied to the assessment process are interventions that emphasize staff's role in helping patients manage strong affects and avoid the use of nonproductive behavior regulation strategies. PMID- 17118055 TI - Top 10 milieu interventions for inpatient child/adolescent treatment. AB - Presented in this article are 10 interventions to deal with children and adolescents on inpatient psychiatric units. The 10 are divided into three categories: behavioral, cognitive, and affective. The interventions discussed are particularly relevant to staff in their efforts to help children and adolescents achieve control over their thoughts, feelings, and behaviors. Each intervention includes a summary of the theory that supports its efficacy, a brief explanation of the intervention, and methods for applying the technique in clinical situations. PMID- 17118056 TI - Mental health needs of children and adolescents at camp: are they being assessed and treated appropriately by the camp nurse? AB - TOPIC: Increasingly, more children and adolescents are attending camps with mental health concerns. This can pose a challenge for camp nurses who may lack experience in assessment and treatment of mental health issues. PURPOSE: To focus on the importance of addressing and treating mental health needs of children and adolescents at camp utilizing the Scope and Standards of Psychiatric Mental Health Nursing Practice. SOURCES: Personal observations, camp nursing experience, and scholarly published literature. CONCLUSIONS: It is paramount that mental health needs of children and adolescents at camp are addressed and managed appropriately by the camp nurse. Education of camp nurses and camp administrators is also a vital part of providing care. PMID- 17118057 TI - The undervalued role of over-regulation in autism: Chaos Theory as a metaphor and beyond. AB - TOPIC: A triad of impairment that consists of impaired communication, impaired social skills and over-regulated behavior characterizes autism. Causality of autism is yet to be identified. Therapy has been devised to apply to the work with children and adolescents with a diagnosis of autism in the domains of impaired communication and impaired social skills. Little attention has been paid to the importance of the over-regulated behavior that forms part of the triad of impairment. PURPOSE: This paper considers this point of over-regulation in the triad of impairment in autism through the lens of Chaos theory. SOURCES: Contemporary literature on autism and Chaos theory. CONCLUSIONS: Implications for nursing practice and research are raised. PMID- 17118059 TI - The petrosquamosal sinus in humans. AB - This article provides a comprehensive description of the morphology of the human petrosquamosal sinus (PSS) derived from original observations made on 13 corrosion casts of the cranial venous system combined with routine clinical imaging studies in two patients. The PSS is not a rare finding in the adult human. In particular, continuous developments in imaging techniques have made radiologists become increasingly aware of this anatomical entity in recent years. The role of the PSS as a major encephalic drainage pathway and its potential implication in pathological conditions such as intracranial venous hypertension are discussed. PMID- 17118060 TI - Mesencephalic human neural progenitor cells transplanted into the neonatal hemiparkinsonian rat striatum differentiate into neurons and improve motor behaviour. AB - Neural stem cell transplantation is a promising strategy for the treatment of neurodegenerative diseases. To evaluate the differentiation potential of human neural progenitor cells (hNPCs) as a prerequisite for clinical trials, we intracerebrally transplanted in vitro expanded fetal mesencephalic hNPCs into hemiparkinsonian rats. On postnatal day one (P1), 17 animals underwent a unilateral intraventricular 6-hydroxydopamine injection into the right lateral ventricle. At P3, animals (n = 10) received about 100,000 hNPCs (1 microL) in the right striatum. Five weeks after birth, animals underwent behaviour tests prior to fixation, followed by immunohistochemistry on brain slices for human nuclei, glial fibrillary acidic protein, S100beta, neuronal nuclei antigen, neuron specific enolase and tyrosine hydroxylase. Compared with the apomorphine-induced rotations in the lesioned-only group (7.4 +/- 0.5 min(-1)), lesioned and successfully transplanted animals (0.3 +/- 0.1 min(-1)) showed a significant therapeutic improvement. Additionally, in the cylinder test, the lesioned-only animals preferred to use the ipsilateral forepaw. Conversely, the lesioned and transplanted animals showed no significant side bias similar to untreated control animals. Transplanted human nuclei-immunoreactive cells were found to survive and migrate up to 2000 microm into the host parenchyma, many containing the pan neuronal markers neuronal nuclei antigen and neuron-specific enolase. In the striatum, tyrosine hydroxylase-immunoreactive somata were also found, indicating a dopaminergic differentiation capacity of transplanted hNPCs in vivo. However, the relative number of tyrosine hydroxylase-immunoreactive neurons in vivo seemed to be lower than in corresponding in vitro differentiation. To minimize donor tissue necessary for transplantation, further investigations will aim to enhance dopaminergic differentiation of transplanted cells in vivo. PMID- 17118061 TI - Chemical coding of myenteric neurons with different axonal projection patterns in the porcine ileum. AB - The aim of this study was to perform an immunohistochemical characterization of two different myenteric neuron types of the pig displaying opposite axonal projections. These were type I neurons equipped with lamellar dendrites that projected mainly orally, and type VI neurons that displayed typical axonal dendrites and projected anally. Double immunostainings of longitudinal muscle/myenteric plexus wholemounts from ileal segments of four pigs were performed to visualize neurofilaments (NF) in combination with calcitonin gene related peptide (CGRP), leu-enkephalin (ENK) and substance P (SP), respectively. Triple immunostainings of wholemounts, using antibodies against neuronal nitric oxide synthase (nNOS) and vasoactive intestinal peptide (VIP) as well as against VIP and galanin (GAL), were performed. We found that 78% of type I neurons immunoreacted to ENK, 21% to CGRP and 24% to SP. The NF-positive type I neurons co-reactive for one of the three above markers displayed mostly frayed outlines of both their somal contours and their broadened dendritic endings. By contrast, most of the non-coreactive type I neurons displayed rather sharply outlined somata and dendrites. No type I neuron immunoreacted to nNOS, VIP or GAL and none of the type VI NF-reactive neurons reacted to CGRP, ENK or SP. All type VI neurons investigated displayed immunoreactivity for nNOS, 92% of which were co reactive for VIP. Co-reactivity for VIP and GAL was found in 69% of type VI neurons, 21% were positive for VIP but negative for GAL, 9% were negative for both GAL and VIP, and 1% were positive for GAL but negative for VIP. We conclude that there are two subpopulations of morphological type I neurons. One of these displays mainly oral projections and could not be further characterized in this study. The other, which may correspond to neurons innervating the longitudinal and circular muscle layers, were partly immunoreactive for ENK, CGRP and/or SP. Type VI neurons are immunoreactive for nNOS frequently co-localized with VIP and, partly, also GAL. These may be inhibitory motor neurons and are different from VIP/GAL-coreactive minineurons described earlier. PMID- 17118062 TI - Ureteric branching morphogenesis in BMP4 heterozygous mutant mice. AB - Exogenous bone morphogenetic protein 4 (BMP4) inhibits ureteric branching morphogenesis and amplifies the already existing branching asymmetry in the developing mouse kidney in vitro. In the present study we examined ureteric branching morphogenesis in BMP4/lacZ heterozygous (BMP4(+/-)) mice in vitro under control conditions and in the presence of exogenous BMP4 using three-dimensional image analysis software. The relative expression of BMP4 mRNA was determined in BMP4(+/-) and wildtype urogenital ridges using real-time PCR. Embryonic day 12.5 (E12.5) BMP4(+/-) and wildtype mouse metanephroi were cultured for 48 h with or without 260 ng mL(-1) recombinant human BMP4 (rhBMP4) and were then wholemount immunostained in order to identify the ureteric epithelium, which was quantified in three dimensions. Despite a significant reduction in BMP4 mRNA in BMP4(+/-) mice, qualitative and quantitative studies identified no differences in ureteric branching morphogenesis between phenotypically normal BMP4(+/-) and wildtype metanephroi in either BMP4-treated or control cultures. Both BMP4(+/-) and wildtype metanephroi cultured in the presence of BMP4 showed a decrease in total ureteric length, branch number and ureteric volume, and increased average branch length compared with control cultures. A marked anterior-posterior asymmetry in both ureteric length, branch number and average branch length was observed in BMP4-treated metanephroi from both genotypes. A similar asymmetry was revealed in control metanephroi from both genotypes. This asymmetry is the result of reduced ureteric branching morphogenesis but not elongation in the posterior region of the kidney. These results suggest that despite reduced endogenous BMP4 mRNA levels, most BMP4(+/-) embryos can still facilitate normal ureteric branching morphogenesis during development. In addition, reduced endogenous levels of BMP4 do not alter the inhibitory effects of exogenous BMP4 on ureteric branching or amplification of normal renal asymmetry. PMID- 17118063 TI - Implications of the functional anatomy of the hand and forearm of Ailurus fulgens (Carnivora, Ailuridae) for the evolution of the 'false-thumb' in pandas. AB - Both the giant panda (Ailuropoda melanoleuca) and the red panda (Ailurus fulgens) possess a 'false-thumb', actually an enlarged radial sesamoid bone, which contributes to the gripping action of the hand. These species are not closely related, however, as one is an ursid and the other an ailurid, so the fact that they share this adaptation implies a remarkable convergence. We studied the functional anatomy of this structure in the red panda, comparing it with existing descriptions of the grasping mechanism in both pandas. Previous interpretations of the radial sesamoid in Ailurus as a rod-like structure without direct articulation to the wrist bones are inaccurate. There are various important differences between the red panda and the giant panda. In the former, the lesser development of the radial sesamoid, its connection with the flexor retinaculum, the presence of an insertion of the muscle abductor pollicis longus in the first metacarpal, which enhances its supinatory action, and the presence of a muscle flexor brevis digitorum manus point to thin-branch climbing features serving as an exaptation to the more recent role of the red panda hand in the manipulation of bamboo. PMID- 17118064 TI - Muscle architecture and functional anatomy of the pelvic limb of the ostrich (Struthio camelus). AB - The functional anatomy of the pelvic limb of the ostrich (Struthio camelus) was investigated in order to assess musculoskeletal specialization related to locomotor performance. The pelvic limbs of ten ostriches were dissected and detailed measurements of all muscle tendon units of the pelvic limb were made, including muscle mass, muscle length, fascicle length, pennation angle, tendon mass and tendon length. From these measurements other muscle properties such as muscle volume, physiological cross-sectional area (PCSA), tendon cross-sectional area, maximum isometric muscle force and tendon stress were derived, using standard relationships and published muscle data. Larger muscles tended to be located more proximally and had longer fascicle lengths and lower pennation angles. This led to an expected proximal to distal reduction in total muscle mass. An exception to this trend was the gastrocnemius muscle, which was found to have the largest volume and PCSA and also had the highest capacity for both force and power production. Generally high-power muscles were located more proximally in the limb, while some small distal muscles (tibialis cranialis and flexor perforatus digiti III), with short fibres, were found to have very high force generation capacities. The greatest proportion of pelvic muscle volume was for the hip extensors, while the highest capacity for force generation was observed in the extensors of the ankle, many of which were also in series with long tendons and thus were functionally suited to elastic energy storage. PMID- 17118065 TI - The structure of the cushions in the feet of African elephants (Loxodonta africana). AB - The uniquely designed limbs of the African elephant, Loxodonta africana, support the weight of the largest terrestrial animal. Besides other morphological peculiarities, the feet are equipped with large subcutaneous cushions which play an important role in distributing forces during weight bearing and in storing or absorbing mechanical forces. Although the cushions have been discussed in the literature and captive elephants, in particular, are frequently affected by foot disorders, precise morphological data are sparse. The cushions in the feet of African elephants were examined by means of standard anatomical and histological techniques, computed tomography (CT) and magnetic resonance imaging (MRI). In both the forelimb and the hindlimb a 6th ray, the prepollex or prehallux, is present. These cartilaginous rods support the metacarpal or metatarsal compartment of the cushions. None of the rays touches the ground directly. The cushions consist of sheets or strands of fibrous connective tissue forming larger metacarpal/metatarsal and digital compartments and smaller chambers which were filled with adipose tissue. The compartments are situated between tarsal, metatarsal, metacarpal bones, proximal phalanges or other structures of the locomotor apparatus covering the bones palmarly/plantarly and the thick sole skin. Within the cushions, collagen, reticulin and elastic fibres are found. In the main parts, vascular supply is good and numerous nerves course within the entire cushion. Vater-Pacinian corpuscles are embedded within the collagenous tissue of the cushions and within the dermis. Meissner corpuscles are found in the dermal papillae of the foot skin. The micromorphology of elephant feet cushions resembles that of digital cushions in cattle or of the foot pads in humans but not that of digital cushions in horses. Besides their important mechanical properties, foot cushions in elephants seem to be very sensitive structures. PMID- 17118066 TI - In vitro culture of enzymatically isolated chondrons: a possible model for the initiation of osteoarthritis. AB - The aim of this study was to assess whether enzymatically isolated chondrons from normal adult articular cartilage could be used as a model for the onset of osteoarthritis, by comparison with mechanically extracted chondrons from osteoarthritic cartilage. Enzymatically isolated chondrons (EC) were cultured for 4 weeks in alginate beads and agarose gel constructs. Samples were collected at days 1 and 2, and weekly thereafter. Samples were immunolabelled for types II and VI collagen, keratan sulphate and fibronectin and imaged using confocal microscopy. Mechanically extracted chondrons (MC) were isolated, immunohistochemically stained for type VI collagen and examined by confocal microscopy. In culture, EC showed the following characteristics: swelling of the chondron capsule, cell division within the capsule and remodelling of the pericellular microenvironment. This was followed by chondrocyte migration through gaps in the chondron capsule. Four types of cell clusters formed over time in both alginate beads and agarose constructs. Cells within clusters exhibited quite distinct morphologies and also differed in their patterns of matrix deposition. These differences in behaviour may be due to the origin of the chondrocytes in the intact tissue. The behaviour of EC in culture paralleled the range of morphologies observed in MC, which presented as single and double chondrons and large chondron clusters. This preliminary study indicates that EC in culture share similar structural characteristics with MC isolated from osteoarthritic cartilage, confirming that some processes that occur in osteoarthritis, such as pericellular remodelling, take place in EC cultures. The study of EC in culture may therefore provide an additional tool to investigate the mechanisms operating during the initial stages of osteoarthritis. Further investigation of specific osteoarthritic phenotype markers will, however, be required in order to validate the value of this model. PMID- 17118067 TI - Inflammation-responsive focal constrictors in the mouse ear microcirculation. AB - In many capillary exchange beds, blood flow is locally regulated by precapillary sphincter-like activity. In this study, we used intravascular tracers and scanning electron microscopy to investigate precapillary blood flow regulation in the mouse ear. Gelatin ink injections of the normal mouse ear demonstrated 6.8 +/ 2.3 axial vessels with a cutoff of detectable tracer in the early branches: 19 +/- 11 focal constrictions were observed along the 1st to 5th order branches of the axial vessels. A perfusion tracer consisting of biotinylated anti-endothelial lectins (Ricinus Communis Agglutin, Lycopersicon Esculentum and Griffonia Simplicifolia) was circulated for 30 min under physiological conditions. Subsequent enzyme histochemistry demonstrated no significant change in distal perfusion or in the number of focal constrictions (P > 0.05). Furthermore, the focal constrictions were unresponsive to vasodilators such as organic nitrates and prostaglandin E1. By contrast, the presence of oxazolone-induced inflammation resulted in significant and sustained vasodilatation for more than 96 h (P > 0.001). Scanning electron microscopy demonstrated discrete constricting bands morphologically distinct from known precapillary sphincters. These results suggest that these previously unappreciated inflammation-responsive precapillary constrictors regulate capillary recruitment in the mouse ear microcirculation. PMID- 17118068 TI - Immunohistochemical assessment of parafibromin in mouse and human tissues. AB - Parafibromin is a protein encoded by the HRPT2 oncosuppressor gene, whose mutation causes the hyperparathyroidism-jaw tumour syndrome, characterized by the occurrence of parathyroid adenoma or carcinoma, fibro-osseous jaw tumours, and renal neoplastic and non-neoplastic abnormalities. Non-morphological techniques, such as Northern and Western blotting and reverse transcriptase-PCR, indicate that parafibromin is ubiquitously expressed, but extensive immunohistochemical studies have not been performed. To increase our knowledge of the distribution and patterns of expression of parafibromin, we examined its expression and location in many different mouse and human organs by immunohistochemistry. There were no substantial differences in parafibromin expression between mouse and human. We found widespread expression of parafibromin, except in connective tissue, smooth muscle, endothelium and some other types of epithelia (colonic, urinary, tubaric, uterine, thyroid). Heterogeneity of positivity intensity and subcellular location (nuclear, nucleocytoplasmic, cytoplasmic) was found between tissues and cell types, suggesting differential functional involvement of parafibromin. Moreover, higher parafibromin expression was found in cell types, such as hepatocytes, cells of the base of gastric glands, renal cortex tubules and the pars intermedia of the hypophysis, which are characterized by different proliferative capacity, thus indicating that the cellular function of parafibromin may not be reduced only to its anti-proliferative effect. PMID- 17118070 TI - Atrophy and anarchy: Third national survey of nursing skill-mix and advanced nursing practice in ophthalmology. AB - AIMS AND OBJECTIVES: The aims of the study were to investigate the advanced nursing practice and the skill-mix of nurses working in ophthalmology. BACKGROUND: The expansion of new nursing roles in the United Kingdom in the past decade is set against the background of a nursing shortage. The plan to modernize the National Health Service and improve the efficiency and delivery of healthcare services as well as to reduce junior doctors' hours contributes towards a profusion of new and more specialized and advanced nursing roles in various areas of nursing including ophthalmology. DESIGN: A self-reporting quantitative questionnaire was employed. The study used comparative and descriptive statistical tests. METHOD: The questionnaires were distributed to all ophthalmic hospitals and units in the United Kingdom. Hospital and unit managers were responsible for completing the questionnaires. RESULTS: Out of a total 181 questionnaires 117 were returned. There is a downward trend in the total number of nurses working in ophthalmology. The results demonstrate more nurses working at an advanced level. However, there is a general confusion regarding role interpretation at the advanced level of practice, evident through the wide range of job titles being used. There was inconsistency in the qualifications expected of these nurses. CONCLUSION: Whilst there are more nurses working at an advanced level this is set against an ageing workforce and an overall decline in the number of nurses in ophthalmology. There is inconsistency in job titles, grades, roles and qualifications for nurses who work at an advanced or higher level of practice. The Agenda for Change with its new structure for grading jobs in the United Kingdom may offer protection and consistency in job titles, pay and qualifications for National Health Service nurse specialists. The Nursing and Midwifery Council needs to provide clear guidelines to the practitioners on educational and professional requirements, to protect patients and nurses. RELEVANCE TO CLINICAL PRACTICE: The findings indicate that there is a need for better regulations for nurses working at advanced nursing practice. PMID- 17118071 TI - Clinical judgement in the interpretation of evidence: A Bayesian approach. AB - AIM: This paper presents an argument for the use of Bayesian reasoning in considering the value of evidence in making nursing judgements. BACKGROUND: Nursing has taken on board the drive towards evidence-based practice. There has been little discussion, however, of how evidence should be interpreted. There is a growing interest in health care in the use of Bayesian reasoning for evidence interpretation, both in research and in clinical practice; as yet, there is a limited discussion in the literature of relevance to nursing. OBJECTIVES: To provide a short tutorial in the application of Bayes rule to a clinical judgement. To discuss the implications for practice of adopting a Bayesian perspective. DISCUSSION: The relationship between evidence and clinical judgement is outlined. The need to accept uncertainty, and be equipped to deal adequately with this, is discussed: some basic ideas of probability are rehearsed. An outline of Bayesian reasoning is offered and a demonstration of the application of Bayes rule to a nursing judgement is presented. RELEVANCE TO PRACTICE: A rationale for adopting a Bayesian perspective on evidence interpretation is offered: namely the changing context of practice, with the blurring of professional boundaries and the need to articulate judgements, the avoidance of error and the opportunity to identify the appropriate areas for investigation in nursing. PMID- 17118072 TI - Patient participation in clinical decision-making in nursing: A comparative study of nurses' and patients' perceptions. AB - AIM AND OBJECTIVES: The aim of this study was to compare the degree of concordance between patients and Registered Nurses' perceptions of the patients' preferences for participation in clinical decision-making in nursing care. A further aim was to compare patients' experienced participation with their preferred participatory role. BACKGROUND: Patient participation in clinical decision-making is valuable and has an effect on quality of care. However, there is limited knowledge about patient preferences for participation and how nurses perceive their patients' preferences. METHODS: A comparative design was adopted with a convenient sample of 80 nurse-patient dyads. A modified version of the Control Preference Scale was used in conjunction with a questionnaire developed to elicit the experienced participation of the patient. RESULTS: A majority of the Registered Nurses perceived that their patients preferred a higher degree of participation in decision-making than did the patients. Differences in patient preferences were found in relation to age and social status but not to gender. Patients often experienced having a different role than what was initially preferred, e.g. a more passive role concerning needs related to communication, breathing and pain and a more active role related to activity and emotions/roles. CONCLUSIONS: Registered Nurses are not always aware of their patients' perspective and tend to overestimate patients' willingness to assume an active role. Registered Nurses do not successfully involve patients in clinical decision making in nursing care according to their own perceptions and not even to the patients' more moderate preferences of participation. RELEVANCE TO CLINICAL PRACTICE: A thorough assessment of the individual's preferences for participation in decision-making seems to be the most appropriate approach to ascertain patient's involvement to the preferred level of participation. The categorization of patients as preferring a passive role, collaborative role or active role is seen as valuable information for Registered Nurses to tailor nursing care. PMID- 17118073 TI - Student nurses' knowledge, skills and attitudes towards the use of portfolios in a school of nursing. AB - AIMS: The aims of this study are to examine the knowledge, skills and attitudes of student nurses about the value and purpose of portfolios and their relevance for professional development. BACKGROUND: Portfolios have become an integral aspect of student centred learning within nursing. The literature is generally positive about the benefits of portfolios but questions remain about their effective implementation. METHOD: A questionnaire was used to survey student nurses about the use of portfolios as a learning tool. The questionnaire was developed specifically for this study and was structured around a knowledge skills and attitude framework. RESULTS: The sample was 413 preregistration student nurses in one school of nursing in the UK across course, year of study and branch of nursing. The main findings reveal that students are unclear but positive about the purpose and use of portfolios. Specifically, academic tutors do not provide sufficient direction in the development of their portfolios. There is also evidence that students found portfolios more useful for exploring the 'art' of nursing, rather than the 'technical' skills of nursing. Students also reported that they would welcome a dedicated portfolio module. CONCLUSION: Students struggle with what is useful about portfolios and are unsure about its purpose. Nonetheless, they are positive towards portfolios as a learning tool and seem to acknowledge the usefulness of portfolios for professional development. RELEVANCE TO CLINICAL PRACTICE: The importance of portfolios for professional development is now being emphasized but how academic tutors and clinical mentors support their development remains unclear. This study provides understanding about what support students would value. PMID- 17118074 TI - Workforce issues in nursing in Queensland: 2001 and 2004. AB - AIM AND OBJECTIVES: The aim of the study was to identify the factors having an impact upon nursing work and to use the results to inform strategic planning of the Queensland Nurses Union. BACKGROUND: In 2001 and 2004, a study was undertaken to gather data on the level of satisfaction of nurses with their working life. This paper reports the 2004 results on workload, skill mix, remuneration and morale. Where applicable, the results are compared with 2001 data. METHODS: A questionnaire was mailed to 3000 Assistants-in-Nursing, Enrolled and Registered Nurses in October 2004. All participants were members of the Queensland Nurses Union. The results are reported in three sectors - public, private and aged care. A total of 1349 nurses responded to the survey, a response rate of 45%. RESULTS: Nurses in the 2004 study believed: their workload was heavy; their skills and experience poorly rewarded; work stress was high; morale was perceived to be poor and, similar to 2001, deteriorating; the skill mix was often inadequate; and the majority of nurses were unable to complete their work in the time available. Nursing morale was found to be associated with autonomy, workplace equipment, workplace safety, teamwork, work stress, the physical demand of nursing work, workload, rewards for skills and experience, career prospects, status of nursing and remuneration. CONCLUSIONS: Overall the findings of the study are consistent with those determined by the 2001 survey. RELEVANCE TO CLINICAL PRACTICE: The findings of this study indicate the importance of factors such as workplace autonomy, teamwork, the levels of workplace stress, workload and remuneration on nursing morale. The data also indicate that workplace safety and workplace morale are linked. These findings provide information for policy makers and nurse managers on areas that need to be addressed to retain nurses within aged care, acute hospital and community nursing. PMID- 17118075 TI - From expert to tasks, expert nursing practice redefined? AB - AIM: The aim of this paper was to explore the concept of expertise in nursing from the perspective of how it relates to current driving forces in health care in which it discusses the potential barriers to acceptance of nursing expertise in a climate in which quantification of value and cost containment run high on agendas. BACKGROUND: Expert nursing practice can be argued to be central to high quality, holistic, individualized patient care. However, changes in government policy which have led to the inception of comprehensive guidelines or protocols of care are in danger of relegating the 'expert nurse' to being an icon of the past. Indeed, it could be argued that expert nurses are an expensive commodity within the nursing workforce. Consequently, with this change to the use of clinical guidelines, it calls into question how expert nursing practice will develop within this framework of care. METHOD: The article critically reviews the evidence related to the role of the Expert Nurse in an attempt to identify the key concepts and ideas, and how the inception of care protocols has implications for their role. CONCLUSION: Nursing expertise which focuses on the provision of individualized, holistic care and is based largely on intuitive decision making cannot, should not be reduced to being articulated in positivist terms. However, the dominant power and decision-making focus in health care means that nurses must be confident in articulating the value of a concept which may be outside the scope of knowledge of those with whom they are debating. RELEVANCE TO CLINICAL PRACTICE: The principles of abduction or fuzzy logic may be useful in assisting nurses to explain in terms which others can comprehend, the value of nursing expertise. PMID- 17118076 TI - Clinical nurse specialists as entrepreneurs: constrained or liberated. AB - AIMS: This qualitative study explored the experiences of two groups of clinical nurse specialists--continence advisors and tissue viability nurses--working in primary care in the UK. In particular, the study focused on how clinical nurse specialists' relationships with other health-care professionals had an impact on their role. BACKGROUND: Clinical nurse specialists are recognized worldwide as having expertise in a given field, which they use to develop the practice of others. Additionally, clinical nurse specialists share many of the characteristics of entrepreneurs, which they use to develop services related to their speciality. However, little research has been conducted in relation to clinical nurse specialists' experiences as they attempt to diversify nursing practice. DESIGN/METHODS: An ethnographic approach was adopted comprising many elements of Glaserian grounded theory. Data were collected via participant observation and face-to-face interviews with 22 clinical nurse specialists. FINDINGS: Services provided by clinical nurse specialists were not static, clinical nurse specialists being the main drivers for service developments. However, clinical nurse specialists encountered difficulties when introducing new ideas. Given their role as advisors, clinical nurse specialists lacked authority to bring about change and were dependent on a number of mechanisms to bring about change, including 'cultivating relationships' with more powerful others, most notably the speciality consultant. CONCLUSIONS: The UK government has pledged to 'liberate the talents of nurses' so that their skills can be used to progress patient services. This study highlights the fact that a lack of collaborative working practices between health-care professionals led to clinical nurse specialists being constrained. RELEVANCE TO CLINICAL PRACTICE: Health-care organizations need to provide an environment in which the entrepreneurial skills of clinical nurse specialists may be capitalized on. In the absence of an outlet for their ideas regarding service developments, clinical nurse specialists may remain dependent on the mechanisms witnessed in this study for some time. PMID- 17118077 TI - In search of details of patient teaching in nursing documentation--an analysis of patient records in a medical ward in Sweden. AB - AIM: The aim of this study was to identify terms and expressions indicating patients' need for knowledge and understanding, as well as nurses' teaching interventions, as documented in nursing records. BACKGROUND: Previous international studies have shown that nursing documentation is often deficient in terms of recording patient teaching. METHODS: Patient records (N = 35) were collected in a general medical ward in a hospital in Sweden. The data contain 206 days of nursing documentation. The records were analysed with regard to content and structure. Terms and expressions indicating patients' need for knowledge and understanding and terms and expressions indicating nurses' teaching activities were analysed. RESULTS: The results showed that patients' need for knowledge is implicitly indicated by conceptions and experiences as well as questions. Furthermore, nurses' implicit teaching interventions consist of information, motivating conversations, explanations, instructions and setting expectations. However, the content and structure of the pedagogical activities in the patient records are fragmented and vague. RELEVANCE TO CLINICAL PRACTICE: Efforts must be directed towards elaborating upon the above-mentioned terms and expressions as indications of patients' need for knowledge and nurses' teaching interventions. Moreover, these terms and expressions must be recognized and acknowledged. PMID- 17118078 TI - The motivation of health professionals to explore research evidence in their practice: An intervention study. AB - AIM: To assess the impact of multifaceted clinically focused educational strategies that concentrated on introducing dementia care research evidence on health professionals' awareness and inclination to use research findings in their future practice. BACKGROUND: The promise of evidence-based practice is slow to materialize with the limitations of adopting research findings in practice readily identifiable. METHOD: A pre- and post-test quasi experimental design. The study involved the administration of: a pretest (baseline), an intervention phase, and a post-test survey, the same research utilization survey. TOOL: The Edmonton Research Orientation Survey (EROS), a self-report tool that asks participants about their attitudes toward research and about their potential to use research findings, was used to determine health professionals' orientation to research. INTERVENTION: The introduction of dementia care research evidence through multifaceted clinically focused educational strategies to improve practice. This was achieved through a resource team comprising a Clinical Nurse Consultant, as a leader and resource of localized evidence-based knowledge in aged care; an experienced Registered Nurse to support the introduction of strategies and a further experienced educator and clinician to reinforce the importance of evidence in change. RESULTS: Across all the four subscales that are measured in the Edmonton Research Orientation Survey, statistical analysis by independent samples t-test identified that there was no significant change between the before and after measurements. RELEVANCE TO CLINICAL PRACTICE: Successful integration of changes based on evidence does not necessarily mean that staff become more aware or are more inclined to use research findings in future to address problems. PMID- 17118079 TI - Caring for patients of Islamic denomination: Critical care nurses' experiences in Saudi Arabia. AB - AIM: To describe the critical care nurses' experiences in caring for patients of Muslim denomination in Saudi Arabia. BACKGROUND: Caring is known to be the essence of nursing but many health-care settings have become more culturally diverse. Caring has been examined mainly in the context of Western cultures. Muslims form one of the largest ethnic minority communities in Britain but to date, empirical studies relating to caring from an Islamic perspective is not well documented. Research conducted within the home of Islam would provide essential truths about the reality of caring for Muslim patients. DESIGN: Phenomenological descriptive. Methods. Six critical care nurses were interviewed from a hospital in Saudi Arabia. The narratives were analysed using Colaizzi's framework. RESULTS: The meaning of the nurses' experiences emerged as three themes: family and kinship ties, cultural and religious influences and nurse patient relationship. The results indicated the importance of the role of the family and religion in providing care. In the process of caring, the participants felt stressed and frustrated and they all experienced emotional labour. Communicating with the patients and the families was a constant battle and this acted as a further stressor in meeting the needs of their patients. CONCLUSIONS: The concept of the family and the importance and meaning of religion and culture were central in the provision of caring. The beliefs and practices of patients who follow Islam, as perceived by expatriate nurses, may have an effect on the patient's health care in ways that are not apparent to many health-care professionals and policy makers internationally. RELEVANCE TO CLINICAL PRACTICE: Readers should be prompted to reflect on their clinical practice and to understand the impact of religious and cultural differences in their encounters with patients of Islam denomination. Policy and all actions, decisions and judgments should be culturally derived. PMID- 17118080 TI - Validation of the Chinese version of the satisfaction with the nursing home instrument. AB - AIM: To assess the psychometric properties of the Chinese version of the Satisfaction with the Nursing Home Instrument. BACKGROUND: Resident's satisfaction has been regarded by the literature as a gold standard for quality of nursing home care. Accurate assessment of resident's satisfaction can provide valuable information for implementation of quality nursing home care. However, there is not a validated Chinese tool to serve the purpose. DESIGN: A cross sectional descriptive survey design. METHODS: Content validity of the Chinese version of the Satisfaction with the Nursing Home Instrument was assessed by the use of expert panel. Construct validity of the Chinese version of the Satisfaction with the Nursing Home Instrument was determined by assessing the correlation between satisfaction with other theoretically related constructs. Internal consistency and stability of the Chinese version of the Satisfaction with the Nursing Home Instrument were determined by Cronbach's method and two week test-retest reliability. The six-factor structure of the Chinese version of the Satisfaction with the Nursing Home Instrument was assessed by confirmatory factor analysis. Testing was performed on a cluster sample of 330 residents from 16 nursing homes in Hong Kong. RESULTS: The Chinese version of the Satisfaction with the Nursing Home Instrument demonstrated good content validity by having content validity index of 0.93. High construct validity of the Chinese version of the Satisfaction with the Nursing Home Instrument was supported by its significant correlation with depression (r = -0.42, P = 0.000), health-related quality of life (physical component) (r = 0.16, P = 0.042), health-related quality of life (mental component) (r = 0.41, P = 0.000) and global quality of care (r = 0.49, P = 0.000). The Chinese version of the Satisfaction with the Nursing Home Instrument demonstrated satisfactory internal consistency and good stability by having Cronbach's alpha of 0.79 and intra-class correlation coefficient of 0.94, respectively. The six-factor structure of the Chinese version of the Satisfaction with the Nursing Home Instrument was not fully supported by confirmatory factor analysis. CONCLUSIONS: The Chinese version of the Satisfaction with the Nursing Home Instrument is a useful instrument for assessing satisfaction of cognitively intact Chinese nursing home residents. Findings provided initial evidence on its validity and reliability. Further empirical testing is recommended to explore its factor structure. RELEVANCE TO CLINICAL PRACTICE: The Chinese version of the Satisfaction with the Nursing Home Instrument can provide guidance to enhance delivery of high-quality nursing home care for the Chinese population. PMID- 17118081 TI - Lifestyle change after elective percutaneous transluminal coronary angioplasty: An Australian perspective. PMID- 17118082 TI - Commentary on He H-G, Polkki T, Pietila A-M and Vehvilainen-Julkunen K (2005). A survey of Chinese nurses' guidance to parents in children's postoperative pain relief. Journal of Clinical Nursing 14, 1075-1082. PMID- 17118084 TI - Commentary on Naumanen P (2006). The health promotion model as assessed by ageing workers. Journal of Clinical Nursing 15, 219-266. PMID- 17118086 TI - Commentary on Purssell E (2005). Symptoms in the host: infection and treatment model. Journal of Clinical Nursing 14, 555-561. PMID- 17118091 TI - Assessing the trade-offs between crossover and parallel group designs in sleep research. AB - Sleep researchers invariably struggle with decisions regarding the optimal design for their studies. Whether such studies involve treatment for insomnia, obstructive sleep apnea, or any other sleep disorder, questions arise regarding the respective trade-offs between a parallel group and a crossover design. This study analyzed the variance structure of commonly measured polysomnographic variables in an effort to describe the statistical impact of these alternate designs. The study examined the effects of opioids on sleep and employed multiple crossovers between placebo, MS-contin, and methadone using a double-blind, randomized crossover design. Thirty-seven healthy subjects were studied. Four of the subjects were unable to complete the protocol for a variety of reasons, and polysomnogram data was unavailable for one subject. Data from 37 subjects provide the basis for this analysis. Despite dropouts, the crossover study was approximately four times as efficient as the parallel group design in terms of being able to recognize differences in deep sleep across these conditions. Other polysomnographic variables also favored the crossover design to varying extents. Despite the operational complexity of a crossover design, the statistical efficiency of this approach makes it a preferable approach for designing intervention studies in sleep research. PMID- 17118092 TI - The effects of chewing versus caffeine on alertness, cognitive performance and cardiac autonomic activity during sleep deprivation. AB - Chewing has been shown to alleviate feelings of sleepiness and improve cognitive performance during the day. This study investigated the effect of chewing on alertness and cognitive performance across one night without sleep as well as the possible mediating role of cardiac autonomic activity. Fourteen adults participated in a randomized, counterbalanced protocol employing a chewing, placebo and caffeine condition. Participants completed tasks assessing psychomotor vigilance, tracking, grammatical reasoning, alertness and sleepiness each hour across the night. All participants received either placebo or caffeine (200 mg), while the chewing condition also chewed on a tasteless and odorless substance for 15 min each hour. Heart rate (HR), root mean square of the successive differences in R-R intervals on the ECG (RMSSD), and preejection period (PEP) were simultaneously recorded. Alertness and cognitive performance amongst the chewing condition did not differ or were in fact worse when compared with placebo. Similarly, measures of HR and RMSSD remained the same between these two conditions; however, PEP was reduced in the later part of the night in the chewing condition compared with a relative increase for placebo. Caffeine led to improved speed and accuracy on cognitive tasks and increased alertness when compared with chewing. Relative increases in RMSSD and reductions in HR were demonstrated following caffeine; however, no change in PEP was seen. Strong associations between cardiac parasympathetic activity and complex cognitive tasks, as well as between subjective alertness and simpler cognitive tasks, suggest a differential process mediating complex versus simple cognitive performance during sleep deprivation. PMID- 17118093 TI - Total sleep deprivation can increase vestibulo-ocular responses. AB - The effect of sleep deprivation on the vestibular function is largely unknown. Some studies have found that postural balance or vestibular reflexes are decreased in sleep-deprived subjects while others found no change. The aim of this study was to evaluate the effect of sleep deprivation on the vestibulo ocular reflex (VOR). Horizontal eye movements were recorded in healthy subjects during earth vertical axis rotation in darkness once after an ordinary night sleep and once after 26-29 h of sleep deprivation. In the first experiment (n = 8), for which rotation was a 60 degrees s(-1) velocity step, sleep deprivation induced a significant increase in VOR gain. In the second experiment (n = 12), for which rotation was sinusoidal (0.2 Hz +/- 25 degrees s(-1)), sleep deprivation induced no significant modification in VOR gain. The difference between the two studies was the abrupt onset of the step stimulation in comparison with the sinusoidal rotation. Because of its unexpected onset and the potential threat to postural balance, the step stimulation may activate the system specialized in reorienting attention towards salient or behaviourally relevant events. This system includes the right temporoparietal cortex, an area also involved in VOR control. A number of studies have found that sleep deprivation alters the activity of this cortical area during attentional tasks. It is therefore our hypothesis that the difference between the effects of these two vestibular stimulations results from a sleep deprivation-induced modulation of the right temporoparietal cortex. PMID- 17118094 TI - Age and individual variability in performance during sleep restriction. AB - The effect of sleep loss on reaction time (RT) performance varies as a function of age, with RTs of older subjects typically showing less decrement (relative to rested baseline) than those of younger subjects. In the current paper, we examined the nature of this relationship in a 7-day sleep restriction study. The number of repeated measures made it possible to model both intra-individual trajectories over days and individual differences in these trajectories. Results revealed (a) consistent individual differences in RT patterns over time after controlling for experimental design effects; (b) less cumulative RT decline among older individuals regardless of the degree of sleep restriction; and (c) consistent individual variability in performance patterns even after accounting for the effects of age. PMID- 17118095 TI - Differential effects of chronic partial sleep deprivation and stress on serotonin 1A and muscarinic acetylcholine receptor sensitivity. AB - Disrupted sleep and stress are often linked to each other, and considered as predisposing factors for psychopathologies such as depression. The depressed brain is associated with reduced serotonergic and enhanced cholinergic neurotransmission. In an earlier study, we showed that chronic sleep restriction by forced locomotion caused a gradual decrease in postsynaptic serotonin-1A receptor sensitivity, whilst chronic forced activity alone, with sufficient sleep time, did not affect receptor sensitivity. The first aim of the present study was to examine whether the sleep loss-induced change in receptor sensitivity is mediated by adrenal stress hormones. The results show that the serotonin-1A receptor desensitization is independent of adrenal hormones as it still occurs in adrenalectomized rats. The second aim of the study was to establish the effects of sleep restriction on cholinergic muscarinic receptor sensitivity. While sleep restriction affected muscarinic receptor sensitivity only slightly, forced activity significantly hypersensitized the muscarinic receptors. This hypersensitization is because of the stressful nature of the forced activity protocol as it did not occur in adrenalectomized rats. Taken together, these data confirm that sleep restriction may desensitize the serotonin-1A receptor system. This is not a generalized effect as sleep restriction did not affect the sensitivity of the muscarinic cholinergic receptor system, but the latter was hypersensitized by stress. Thus, chronic stress and sleep loss may, partly via different pathways, change the brain into a direction as it is seen in mood disorders. PMID- 17118096 TI - A phylogenetic analysis of the correlates of sleep in birds. AB - Quantitative comparative studies of sleep have focused exclusively on mammals. Such studies have repeatedly found strong relationships between the time spent in various sleep states and constitutive variables related to morphology, physiology, and life history. These studies influenced the development of several prominent hypotheses for the functions of sleep, but the applicability of these patterns and hypotheses to non-mammalian taxa is unclear. Here, we present the first quantitative analysis of sleep in a non-mammalian taxon (birds), focusing on the daily amount of time spent in slow-wave sleep (SWS) and rapid-eye movement (REM) sleep as determined by electrophysiological methods. We examined the relationships between constitutive and sleep variables in 23 avian species following earlier studies in mammals, but also considered an index of exposure to predators while asleep and controlled for shared evolutionary history among taxa. Overall, our results were very different from those obtained for mammals. Most remarkably, the relationships between both SWS time and REM sleep time and all constitutive variables were very weak and markedly non-significant, even though we had adequate power to detect correlations typical of the mammalian data. Only an index of exposure to predation during sleep was significantly related to sleep time, which is the only result common to both birds and mammals. Our results suggest that further insight into the function(s) of sleep across the animal kingdom may require an expansion of sleep research beyond the current mammalian paradigm. PMID- 17118097 TI - Prevalence of sleep problems and their association with inattention/hyperactivity among children aged 6-15 in Taiwan. AB - This study investigated the 6-month prevalence rates of sleep-related problems and their association with daytime inadvertent napping, inattention, hyperactivity/impulsivity, and oppositional symptoms in children and adolescents. A representative school-based sample of 2463 first to ninth graders was recruited using a multistage sampling method. The instruments included the Sleep Habits Questionnaire (including dyssomnia, parasomnia, sleep schedules, and sleep disordered breathing), the Chinese Health Questionnaire, and the Chinese versions of the Conners' Parent and Teacher Rating Scales-Revised: Short forms. The informants were mothers and teachers. The linear and nonlinear mixed models were used for statistical analyses and sex and age were controlled in the model. Results showed that the rates of middle insomnia, disturbed circadian rhythm, mouth breathing, and daytime inadvertent napping increased with age; whereas those of bedwetting, bruxism, sleep terrors decreased with age. Dyssomnia, sleep disordered breathing problems, daytime inadvertent napping, and sleep schedules were related to attention-deficit/hyperactivity disorder (ADHD)-related symptoms as assessed by mothers' and teachers' ratings. Parasomnia was associated with ADHD-related symptoms as assessed by mothers' ratings. Our findings suggest an age trend of sleep problems similar to those found in the literature and the association of daytime inadvertent napping, inattention, hyperactivity/impulsivity, and oppositional symptoms with sleep-related problems. PMID- 17118098 TI - Peripheral electrical nerve stimulation and rest-activity rhythm in Alzheimer's disease. AB - Rest-activity rhythm disruption is a prominent clinical feature of Alzheimer's disease (AD). The origin of the altered rest-activity rhythm is believed to be degeneration of the suprachiasmatic nucleus (SCN). In accordance with the 'use it or lose it' hypothesis of Swaab [Neurobiol Aging 1991, 12: 317-324] stimulation of the SCN may prevent age-related loss of neurons and might reactivate nerve cells that are inactive but not lost. Previous studies with relatively small sample sizes have demonstrated positive effects of peripheral electrical nerve stimulation on the rest-activity rhythm in AD patients. The present randomized, placebo-controlled, parallel-group study was meant to replicate prior findings of electrical stimulation in AD in a substantially larger group of AD patients. The experimental group (n = 31) received peripheral electrical nerve stimulation and the placebo group (n = 31) received sham stimulation. Effects of the intervention on the rest-activity rhythm were assessed by using wrist-worn actigraphs. Near significant findings on the rest-activity rhythm partially support the hypothesis that neuronal stimulation enhances the rest-activity rhythm in AD patients. Interestingly, post-hoc analyses revealed significant treatment effects in a group of patients who were not using acetylcholinesterase inhibitors concomitantly. We conclude that more research is needed before firm general conclusions about the effectiveness of electrical stimulation as a symptomatic treatment in AD can be drawn. In addition, the present post-hoc findings indicate that future studies on non-pharmacological interventions should take medication use into account. PMID- 17118099 TI - N-terminal pro-brain natriuretic peptide for detection of cardiovascular stress in patients with obstructive sleep apnea syndrome. AB - Patients with obstructive sleep apnea syndrome (OSAS) have an elevated incidence of cardiovascular events that may be related to an increased ventricular load and hypoxemia caused by apneas and hypopneas. N-terminal pro-brain natriuretic peptide (NTproBNP) appears to be an excellent marker of myocardial stretch and could serve as an indicator of subclinical cardiac stress, thereby identifying a patient population at risk for cardiac effects from OSAS. Adult patients presenting with suspected OSAS and scheduled for nocturnal polysomnography were recruited. Patients with heart or renal failure or severe lung disease were excluded. NTproBNP was measured the evening before and the morning after sleep. Blood pressure (BP) was monitored intermittently throughout the night. Fifteen male and 15 female subjects with a mean +/- SD body mass index of 38.2 +/- 9.8 were studied. Mean Apnea-Hypopnea Index (AHI) was 38.4 +/- 26, with 17 subjects having severe OSAS (AHI > 30). No subject had a significant rise in BP. NTproBNP values overnight decreased in 19 patients and rose in 11 (mean change 3.8 +/- 33 pg mL(-1)), but only one patient had an abnormal morning value. Three patients had an abnormal NTproBNP value prior to sleep, but their levels decreased with sleep. No correlations were detected between the evening baseline or postsleep NTproBNP levels and OSAS. Monitoring pre- and postsleep NTproBNP levels revealed no association with the occurrence or degree of OSAS, making it unlikely that NTproBNP could serve as a marker of cardiac stress in OSAS patients with stable BP and without overt heart failure. PMID- 17118100 TI - Changes in dreaming induced by CPAP in severe obstructive sleep apnea syndrome patients. AB - To study dream content in patients with severe obstructive sleep apnea syndrome (OSAS) and its modification with Continuous Positive Airway Pressure (CPAP) therapy. We assessed twenty consecutive patients with severe OSAS and 17 healthy controls. Polysomnograms were recorded at baseline in patients and controls and during the CPAP titration night, 3 months after effective treatment and 2 years later in patients. Subjects were awakened 5-10 min after the beginning of the first and last rapid eye movement (REM) sleep periods and we measured percentage of dream recall, emotional content of the dream, word count, thematic units, sleep architecture and REM density. Dream recall in REM sleep was similar in patients at baseline and controls (51.5% versus 44.4% respectively; P = .421), decreased to 20% and 24.3% the first and third month CPAP nights, and increased to 39% 2 years later (P = 0.004). Violent/highly anxious dreams were only seen in patients at baseline. Word count was higher in patients than in controls. REM density was highest the first CPAP night. Severe OSAS patients recall dreams in REM sleep as often as controls, but their dreams have an increased emotional tone and are longer. Despite an increase in REM density, dream recall decreased the first months of CPAP and recovered 2 years later. Violent/highly anxious dreams disappeared with treatment. A dream recall decrease with CPAP is associated with normalization of sleep in OSAS patients. PMID- 17118101 TI - Habitual snoring is associated with elevated hemoglobin A1c levels in non-obese middle-aged adults. AB - Hemoglobin A(1c) (HbA(1c)) is an indicator of long-term glycemic control. The purpose of this study was to determine whether habitual snoring is associated with increased HbA(1c) levels in non-obese and normoglycemic middle-aged men and women. A total of 6981 subjects (3362 men and 3619 women) aged 40-69 years from the Korean Health and Genome Study were examined for the study. Each participant received a comprehensive physical examination as well as a set of questions pertaining to demographic characteristics and snoring frequency. Habitual snoring was defined as a snoring frequency of > or = 4 days week(-1). After adjusting for age, abdominal obesity, and other confounding covariates, male habitual snorers showed a 1.69-fold excess [95% confidence interval (CI) 1.30-2.19] odds of having a high HbA(1c) level. Similarly, premenopausal women with habitual snoring had a 2.31 times (95% CI 1.22-4.39) significantly higher odds of having elevated HbA(1c)levels compared with non-snorers. This association was not found in postmenopausal women. Multivariate analysis revealed that male habitual snorers aged 40-50 had a 2.08-fold excess (95% CI 1.40-3.09) risk of having an elevated HbA(1c) level. In male habitual snores over 50, the strength of association was attenuated. Our findings based on cross-sectional data support a hypothesis that habitual snoring is associated with impaired glucose tolerance even in non-obese and normoglycemic men and premenopausal women. However, as waist circumference as an index of abdominal obesity (visceral adiposity) in the present study may only partially represent the effect of visceral fat, there may be a residual confounding from visceral obesity in our result. Longitudinal follow-up studies are necessary to confirm the association between sleep-disordered breathing and impaired glucose tolerance and to examine the causal relationship in a healthy population without obesity and diabetes. PMID- 17118102 TI - Temazepam at high altitude reduces periodic breathing without impairing next-day performance: a randomized cross-over double-blind study. AB - The aim of the study was to examine the efficacy and safety of temazepam on nocturnal oxygenation and next-day performance at altitude. A double-blind, randomized, cross-over trial was performed in Thirty-three healthy volunteers. Volunteers took 10 mg of temazepam and placebo in random order on two successive nights soon after arrival at 5000 m, following a 17-day trek from 410 m. Overnight SaO(2) and body movements, and next-day reaction time, maintenance of wakefulness and cognition were assessed. Compared with placebo, temazepam resulted in a reduction in periodic breathing from a median (range) of 16 (0 81.3)% of the night to 9.4 (0-79.6)% (P = 0.016, Wilcoxon's signed-rank test), associated with a small but significant decrease in mean nocturnal SaO(2) from 78 (65-84)% to 76 (64-83)% (P = 0.013). There was no change in sleep latency (P = 0.40) or restlessness (P = 0.30). Temazepam had no adverse effect on next-day reaction time [241 (201-380) ms postplacebo and 242 (204-386) ms post-temazepam], maintenance of wakefulness (seven trekkers failed to maintain 40 min of wakefulness postplacebo, and four post-temazepam), cognition or acute mountain sickness. At high altitude temazepam reduces periodic breathing during sleep without an adverse effect on next-day reaction time, maintenance of wakefulness or cognition. The 2% reduction in mean SaO(2) post-temazepam is likely to be predominantly because of acclimatization, as by chance more trekkers took temazepam on the first night (19 versus 14). We conclude that at high altitude temazepam is effective in reducing periodic breathing, and is safe to use, without any adverse effect upon next-day performance. PMID- 17118103 TI - Screening for sleep-related breathing disorders by transthoracic impedance recording integrated into a Holter ECG system. AB - In patients with arrhythmias, coincidence with sleep-related breathing disorders (SRBD) is high and of clinical relevance. Electrocardiogram-derived (ECG) parameters have been developed for SRBD screening, but it has proved necessary to exclude patients with frequent arrhythmias. Holter-based screening tools, easy to use, are therefore warranted. The goal of our study was to evaluate the diagnostic accuracy, with respect to SRBD detection, of transthoracic impedance recording (TTIR) integrated into a Holter System. Our investigation consisted of 2 phases. In phase 1 we compared the performance of TTIR to that of in-hospital polysomnography (PSG) in 56 patients (46 male, mean age 57). In phase 2 we compared TTIR to results from an ambulatory polygraphy (PG) system in 180 patients (143 male, mean age 56). We scored apnea and hypopnea from P(S)G, and derived a respiratory-disturbance index (P(S)G-RDI). TTIR was analyzed semi automatically. Reduction of the impedance amplitude by more than 50% over 10 s was scored as apnea/hypopnea, with consequent calculation of TTIR-RDI. In phase 1, 20 out of 56 patients revealed a PSG-RDI > 10 h(-1). TTIR-RDI in 19 patients from this group was >10 h(-1) (sensitivity 95%, specificity 97.2%, positive predictive value 95%, negative predictive value 97.2%, interclass correlation coefficient 0.98). In phase 2, 46 of 180 patients revealed a PSG-RDI > 10 h(-1). TTIR-RDI in 37 out of this group was >10 h(-1) (sensitivity 80.4%, specificity 92.5%, positive predictive value 78.7%, negative predictive value 93.2%, interclass correlation coefficient 0.92). TTIR integrated into a Holter ECG system and tested in a large patient cohort demonstrates acceptable high accuracy in detection of SRBD. Arrhythmia analysis and screening for SRBD can be performed in a single-step approch. PMID- 17118104 TI - Brain natriuretic peptide is unchanged after 4 weeks of continuous positive airway pressure therapy. PMID- 17118107 TI - The love song of a dean, with apologies to T S Eliot. PMID- 17118108 TI - Are women residency supervisors obligated to nurture? PMID- 17118109 TI - A comprehensive assessment of medical schools in Bosnia and Herzegovina. AB - OBJECTIVES: To perform internal and external evaluations of all 5 medical schools in Bosnia and Herzegovina against international standards. METHODS: We carried out a 2-stage survey study using the same 5-point Likert scale for internal and external evaluations of 5 medical schools in Bosnia and Herzegovina (Banja Luka, Foca/East Sarajevo, Mostar, Sarajevo and Tuzla). Participants consisted of managerial staff, teaching staff and students of medical schools, and external expert assessors. Main outcome measures included scores on internal and external evaluation forms for 10 items concerning aspects of school curriculum and functioning: 'School mission and objectives'; 'Curriculum'; 'Management'; 'Staff'; 'Students'; 'Facilities and technology'; 'Financial issues'; 'International relationships'; 'Internal quality assurance', and 'Development plans'. RESULTS: During internal assessment, schools consistently either overrated their overall functioning (Foca/East Sarajevo, Mostar and Tuzla) or markedly overrated or underrated their performance on individual items on the survey (Banja Luka and Sarajevo). Scores for internal assessment differed from those for external assessment. These differences were not consistent, except for the sections 'School mission and objectives', 'Curriculum' and 'Development plans', which were consistently overrated in the internal assessments. External assessments was more positive than internal assessments on 'Students' and 'Facilities and technology' in 3 of 5 schools. CONCLUSIONS: This assessment exercise in 5 medical schools showed that constructive and structured evaluation of medical education is possible, even in complex and unfavourable conditions. Medical schools in Bosnia and Herzegovina have successfully formed a national consortium for formal collaboration in curriculum development and reform. PMID- 17118110 TI - Diagnostic ability in relation to clinical seminars and extended-matching questions examinations. AB - CONTEXT: At the Katholieke Universiteit Leuven, Belgium, final year medical students participate in 70 problem-solving clinical seminars over a period of 2 months, concentrating on clinical reasoning for formulating differential diagnoses. The clinical seminars end in an examination consisting of 200 extended matching questions (EMQs). OBJECTIVES: This study asks whether problem-solving clinical seminars improve clinical reasoning and whether the EMQ examination measures clinical decision making. In order to detect an increase in diagnostic ability, the Diagnostic Thinking Inventory (DTI) was applied. The research hypotheses were: (i) DTI scores will be higher after the seminars than before, and (ii) the correlations between DTI scores and EMQ examination scores will be significant. METHODS: In the academic year 2003-04, 3 series of problem-solving clinical seminars were held. At the beginning and end of each series the students filled in the DTI. This questionnaire measures 2 aspects of diagnostic thinking: the degree of flexibility in thinking, and how knowledge is structured in the memory. RESULTS: For all data together, the DTI scores after the clinical seminars were significantly higher than before. Pearson correlations between DTI scores and EMQ examination scores were low but significant, with the exception of post-test Structure (not significant). CONCLUSIONS: Two months of intensive problem-solving clinical training was accompanied by an improvement in diagnostic thinking, as measured by DTI scores. Correlation between DTI scores and examination scores indicates that the EMQs measure an aspect of student achievement that is related to clinical reasoning. PMID- 17118111 TI - Evaluating medical students' non-verbal communication during the objective structured clinical examination. AB - OBJECTIVES: Non-verbal communication (NVC) in medical encounters is an important method of exchanging information on emotional status and contextualising the meaning of verbal communication. This study aimed to assess the impact of medical students' NVC on interview evaluations by standardised patients (SPs). METHODS: A total of 89 medical interviews in an objective structured clinical examination (OSCE) for post-clerkship medical students were analysed. All interviews were videotaped and evaluated on 10 non-verbal behaviour items. In addition, the quality of the interview content was rated by medical faculty on 5 items and the interview was rated by SPs on 5 items. The relationships between student NVC and SP evaluation were examined by multivariate regression analyses controlling for the quality of the interview content. RESULTS: Standardised patients were likely to give higher ratings when students faced them directly, used facilitative nodding when listening to their talk, looked at them equally when talking and listening, and spoke at a similar speed and voice volume to them. These effects of NVC remained significant after controlling for the quality of the interview content. CONCLUSIONS: This study provided evidence of specific non-verbal behaviours of doctors that may have additional impacts on the patient's perception of his or her visit, independently of the interview content. Education in basic NVC should be incorporated into medical education alongside verbal communication. PMID- 17118112 TI - Good for them or good for us? The role of academic guidance interviews. AB - CONTEXT: Weak students may pass unnoticed through their undergraduate programme until they fail a summative assessment. Early recognition and support of struggling students is crucial if they are to avoid subsequent failure at high stakes assessments. METHODS: We introduced a whole-class formative assessment 5 months before a summative assessment. Subsequently, we supported poorly performing students with academic guidance interviews and remedial support. RESULTS: The programme was evaluated by questionnaires issued to staff and students involved in the guidance interviews, and by recording the performance of students who had failed the formative assessment in the subsequent summative assessment. CONCLUSIONS: We conclude that the rescue of failing students requires early recognition and tailored intervention, with support for all participants. PMID- 17118113 TI - Evaluation of residency training in the delivery of culturally effective care. AB - OBJECTIVE: To augment resident training in the delivery of culturally effective care in order to improve clinician capacity to effectively care for patients from diverse backgrounds. METHODS: Residents from the Naval Medical Center San Diego and the University of California San Diego participated in experiential learning and service activities. Programme evaluation assessed aspects of the delivery of culturally effective care in community settings. A community-based participatory approach to engaging residents in the delivery of culturally effective care and evaluation of the effectiveness of this approach are described. RESULTS: A significant pre-post rotation increase was noted in residents' self-perceived ability to identify culture-related issues that may impact on the patient's view of illness (P<0.001) and ability to address a culture-related issue (P<0.001). Community evaluations rated residents positively on behaviours that reflected communication skills and professionalism, but less positively on knowledge about communities. CONCLUSIONS: The authors conclude that resident exposure to the block rotation curriculum contributes to improved knowledge of the skills necessary to provide culturally effective care in diverse community settings. PMID- 17118114 TI - Use of critical incidents to develop a rating form for resident evaluation of faculty teaching. AB - CONTEXT: Monitoring the teaching effectiveness of attending physicians is important to enhancing the quality of graduate medical education. METHODS: We used a critical incident technique with 35 residents representing a cross-section of programmes in a teaching hospital to develop a 23-item rating form. We obtained ratings of 11 attending physicians in internal medicine and general surgery from 54 residents. We performed linear and logistic regression analysis to relate the items on the form to the residents' overall ratings of the attending physicians and the programme directors' ratings of the attending physicians. RESULTS: The residents rated the attending physicians highly in most areas, but lower in provision of feedback, clarity of written communication and cost-effectiveness in making clinical decisions. When we used the residents' overall ratings as the criterion, the most important aspects of attending physicians' teaching were clarity of written communication, cost-effectiveness, commitment of time and energy and whether the resident would refer a family member or friend to the physician. When we used the programme directors' ratings as the criterion, the additional important aspects of performance were concern for the residents' professional well-being, knowledge of the literature and the delivery of clear verbal and written communication. CONCLUSIONS: The critical incident technique can be used to develop an instrument that demonstrates content and construct validity. We found that residents consider commitment of time to teaching and clinical effectiveness to be the most important dimensions of faculty teaching. Other important dimensions include written and verbal communication, cost-effectiveness and concern for residents' professional development. PMID- 17118115 TI - Factor instability of clinical teaching assessment scores among general internists and cardiologists. AB - CONTEXT: We are unaware of studies examining the stability of teaching assessment scores across different medical specialties. A recent study showed that clinical teaching assessments of general internists reduced to interpersonal, clinical teaching and efficiency domains. We sought to determine the factor stability of this 3-dimensional model among cardiologists and to compare domain-specific scores between general internists and cardiologists. METHODS: A total of 2000 general internal medicine and cardiology hospital teaching assessments carried out from January 2000 to March 2004 were analysed using principal factor analysis. Internal consistency and inter-rater reliability were calculated. Mean item scores were compared between general internists and cardiologists. RESULTS: The interpersonal and clinical teaching domains previously demonstrated among general internists collapsed into 1 domain among cardiologists, whereas the efficiency domain remained stable. Internal consistency of domains (Cronbach's alpha range 0.89-0.93) and inter-rater reliability of items (range 0.65-0.87) were good to excellent for both specialties. General internists scored significantly higher (P<0.05) than cardiologists on most items except for 4 items that more accurately assessed the cardiology teaching environment. CONCLUSIONS: We observed factor instability of clinical teaching assessment scores from the same instrument administered to general internists and cardiologists. This finding was attributed to salient differences between these specialties' educational environments and highlights the importance of validating assessments for the specific contexts in which they are to be used. Future research should determine whether interpersonal domain scores identify superior teachers and study the reasons why interpersonal and clinical teaching domains are unstable across different educational settings. PMID- 17118116 TI - Pharmacist-supported medication review training for general practitioners: feasibility and acceptability. AB - OBJECTIVES: To investigate the feasibility and acceptability of training for general practitioners (GPs) in medication review provided by practice pharmacists. METHODS: Within the setting of a Scottish Local Health Care Co operative incorporating 17 general practices, practice pharmacists delivered a 1 hour practice-based group training session to interested GPs, covering a systematic approach to medication review and case studies. One session of funded locum cover was provided for each GP to review up to 6 patients of his or her choice. Practice pharmacists and GPs reviewed patient notes together before GPs saw patients alone. Subsequently pharmacists abstracted data from medical records. Medication-related issues identified during reviews and resultant actions were categorised. The views of GPs on the training were obtained by postal questionnaires. RESULTS: Training was received by 51/74 GPs from 10/17 practices. In 174 reviews analysed, differences in patient medication use from computer records, ineffective medication and missing computer diagnoses were identified most frequently. There was a median of 5 actions per patient, including a median of 2 changes in prescribed drugs. Only 3 reviews resulted in no actions, while 80% of patients had at least 1 prescribing record change. A total of 27 (61%) GPs returned questionnaires; most considered medication review important and were satisfied with the training. Confidence in conducting reviews increased in 14 (52%) GPs. Many indicated they would increase reviews, but time was a barrier for almost all. Although few considered contract or accreditation to be motivating factors, most agreed the training would help them achieve standards for both. CONCLUSIONS: Training by pharmacists was feasible and acceptable, but time constraints may limit the translation of reviews into routine practice. PMID- 17118117 TI - Trainee doctors' opinion of Modernising Medical Careers: a questionnaire survey in the London region. PMID- 17118118 TI - Dynamic organization of the actin cytoskeleton during meiosis and spore formation in budding yeast. AB - During sporulation in Saccharomyces cerevisiae, the four daughter cells (spores) are formed inside the boundaries of the mother cell. Here, we investigated the dynamics of spore assembly and the actin cytoskeleton during this process, as well as the requirements for filamentous actin during the different steps of spore formation. We found no evidence for a polarized actin cytoskeleton during sporulation. Instead, a highly dynamic network of non-polarized actin cables is present underneath the plasma membrane of the mother cell. We found that a fraction of prospore membrane (PSM) precursors are transported along the actin cables. The velocity of PSM precursors is diminished if Myo2p or Tpm1/2p function is impaired. Filamentous actin is not essential for meiotic progression, for shaping of the PSMs or for post-meiotic cytokinesis. However, actin is essential for spore wall formation. This requires the function of the Arp2/3p complex and involves large carbohydrate-rich compartments, which may be chitosome analogous structures. PMID- 17118120 TI - Golgin-160 promotes cell surface expression of the beta-1 adrenergic receptor. AB - Golgin-160 is a ubiquitously expressed peripheral Golgi membrane protein that is important for transduction of certain pro-apoptotic signals at the Golgi complex. However, the role of golgin-160 in normal Golgi structure and function is unknown. Here, we show that depletion of golgin-160 using RNA interference (RNAi) does not affect Golgi morphology or constitutive membrane traffic in HeLa cells. However, depletion of golgin-160 leads to significantly decreased cell surface levels of exogenously expressed beta1-adrenergic receptor (beta1AR), which can be rescued by expression of RNAi-resistant forms of golgin-160. Furthermore, overexpression of golgin-160 leads to higher surface levels of beta1AR. Golgin 160 is localized mostly in the cis and medial regions of the Golgi stack by immunoelectron microscopy, suggesting that it does not directly promote incorporation of beta1AR into transport vesicles at the trans Golgi network. Golgin-160 interacts with beta1AR in vitro, and we mapped the interaction to a region between residues 140 and 257 in the head of golgin-160 and the third intracellular loop of beta1AR. Our results support the idea that golgin-160 may promote efficient surface delivery of a subset of cargo molecules. PMID- 17118119 TI - Citron kinase, a RhoA effector, enhances HIV-1 virion production by modulating exocytosis. AB - RhoGTPases play important roles in the regulation of protein transport and membrane recycling. Little is known, however, about how RhoGTPases affect HIV-1 virion production, which is dependent on the endosomal sorting pathway. We report that ectopic expression of citron kinase (citron-K), a RhoA effector, preferentially enhances HIV-1 virion production. Depletion of endogenous citron-K inhibits HIV-1 virion production. Citron-N, which lacks the kinase domain, also enhances HIV-1 virion production. The leucine zipper, Rho-binding and zinc finger domains of citron-N are necessary for the enhancement activity. Citron-K also enhances murine leukemia virion production and the HIV-1 late domain is not required for the citron-K-mediated enhancement. Ectopic expression of citron-K leads to the formation of cytoplasmic structures containing citron-K and HIV-1 Gag proteins. HIV-1 and citron-K cooperatively enhance acidic endosome and lysosome compartments. Finally, citron-K promotes exocytosis of microvesicles or exosomes that co-purify with HIV-1 virions. We conclude that citron-K enhances HIV-1 virion production by stimulating the endosomal compartments and exocytosis. PMID- 17118121 TI - Ratiometric fluorescence-imaging assays of plant membrane traffic using polyproteins. AB - Fluorescent protein markers are widely used to report plant membrane traffic; however, effective protocols to quantify fluorescence or marker expression are lacking. Here the 20 residue self-cleaving 2A peptide from Foot and Mouth Disease Virus was used to construct polyproteins that expressed a trafficked marker in fixed stoichiometry with a reference protein in a different cellular compartment. Various pairs of compartments were simultaneously targeted. Together with a bespoke image analysis tool, these constructs allowed biosynthetic membrane traffic to be assayed with markedly improved sensitivity, dynamic range and statistical significance using protocols compatible with the common plant transfection and transgenic systems. As marker and effector expression could be monitored in populations or individual cells, saturation phenomena could be avoided and stochastic or epigenetic influences could be controlled. Surprisingly, mutational analysis of the ratiometric assay constructs revealed that the 2A peptide was dispensable for efficient cleavage of polyproteins carrying a single internal signal peptide, whereas the signal peptide was essential. In contrast, a construct bearing two signal peptide/anchors required 2A for efficient separation and stability, but 2A caused the amino-terminal moiety of such fusions to be mis-sorted to the vacuole. A model to account for the behaviour of 2A in these and other studies in plants is proposed. PMID- 17118122 TI - Balanced prescribing. PMID- 17118123 TI - Construction and evaluation of a web-based interactive prescribing curriculum for senior medical students. AB - AIMS: To develop and evaluate for the National Prescribing Service (NPS) a web based interactive prescribing curriculum for Australian senior medical students based on the World Health Organization's Guide to Good Prescribing. METHODS: Teachers of prescribing from all Australian medical schools in 2000 wrote 12 case based modules which were converted to on-line format. Objective evidence was provided for selecting first-line medicines from available alternatives by comparing efficacy, safety, convenience and cost. The curriculum was made available to final year students in 2001 and was evaluated by measuring use from web statistics and by semistructured interviews with 15 teachers (2003) and on line surveys of 363 students over 2003 and 2004. RESULTS: By 2004 the curriculum was used by nine of 11 possible medical schools. Uptake increased each year from 2001 and all 12 modules were accessed consistently. Student access was significantly (P < 0.001) greater when prescribing was an assessable part of their course. Teachers' evaluations were uniformly supportive and the curriculum is seen as a valuable resource. Student responses came from a small proportion of those with password access but were also supportive. Over half of student respondents had created their own evidence-based formulary. CONCLUSIONS: A collaborative venture initiated by the NPS with Australian medical schools has been successfully implemented in most courses. Teachers find the resource of high quality. Student respondents find the curriculum valuable in developing their own prescribing skills. It is best delivered by self-directed study followed by tutorial discussion of prescribing decisions. PMID- 17118124 TI - Aliskiren, a novel orally effective renin inhibitor, exhibits similar pharmacokinetics and pharmacodynamics in Japanese and Caucasian subjects. AB - AIMS: Aliskiren is the first in a new class of orally effective renin inhibitors for the treatment of hypertension. This study compared the pharmacokinetic and pharmacodynamic properties of aliskiren in Japanese and Caucasian subjects. METHODS: In this open-label, single-centre, parallel-group, single- and multiple dose study, 19 Japanese and 19 Caucasian healthy young male subjects received a single 300-mg oral dose of aliskiren on day 1 and then aliskiren 300 mg once daily on days 4-10. Blood samples were collected for the measurement of plasma aliskiren concentration, plasma renin concentration (PRC) and plasma renin activity (PRA). RESULTS: Pharmacokinetic parameters were comparable in Japanese and Caucasian subjects following administration of a single dose of aliskiren {ratio of geometric means: C(max) 1.12 [90% confidence interval (CI) 0.88, 1.43]; AUC(0-72 h) 1.19 [90% CI 1.02, 1.39]} and at steady state [mean ratio: C(max) 1.30 (90% CI 1.00, 1.70); AUC(0-tau) 1.16 (90% CI 0.95, 1.41)]. There was no notable difference in the plasma half-life of aliskiren between Japanese and Caucasian groups (29.7 +/- 10.2 h and 32.0 +/- 6.6 h, respectively). At steady state, peak PRC level and AUC for the concentration-time plot were not significantly different between Japanese and Caucasian subjects (P = 0.64 and P = 0.80, respectively). A single oral dose of aliskiren significantly reduced PRA to a similar extent in Japanese and Caucasian subjects (by 87.5% and 85.7%, respectively, compared with baseline; P < 0.01). Aliskiren was well tolerated by both ethnic groups. CONCLUSIONS: The oral renin inhibitor aliskiren demonstrated similar pharmacokinetic and pharmacodynamic properties in Japanese and Caucasian subjects. PMID- 17118125 TI - Predicting sustained virological response and anaemia in chronic hepatitis C patients treated with peginterferon alfa-2a (40KD) plus ribavirin. AB - AIM: To assess the likelihood of a sustained virological response (SVR) vs. the likelihood of anaemia in patients with chronic hepatitis C. METHODS: Data from 1732 patients treated with peginterferon alfa-2a (40KD) plus ribavirin in two randomized, multinational studies were pooled. Probabilities of SVR and anaemia were modelled using the generalized additive logistic model, with numerous clinical variables considered for entry into the model. Baseline haemoglobin was only considered in the analysis for anaemia. RESULTS: The probability of anaemia increased from 6 to 16% as a function of the ribavirin dose kg(-1) (12-16 mg kg( 1)), whereas the relationship between SVR and ribavirin dose kg(-1) was influenced by hepatitis C virus (HCV) genotype. The probability of an SVR was not influenced by the ribavirin dose kg(-1) in patients with HCV genotype 2 or 3 infection, but increased as a function of ribavirin dose kg(-1) in patients with HCV genotype 1 infection (40-50% increase in probability of SVR for 12-16 mg kg( 1) dose ribavirin increase). The probability of an SVR in patients included with HCV genotype 1 decreased with increasing HCV RNA level to about 3 million copies ml(-1), but was relatively independent of increasing HCV RNA level thereafter. In addition, older age, a higher ribavirin apparent oral clearance and cirrhosis had a negative impact on achieving an SVR, but improved with increasing alanine aminotransferase (ALT) quotient. Sex and ribavirin dose kg(-1) were the most important prognostic factors for anaemia, followed by baseline haemoglobin, age, baseline ALT quotient and cirrhosis. CONCLUSION: This study supports individualizing ribavirin dosages by HCV genotype and body weight, and highlights several clinical variables that influence the likelihood of an SVR compared with anaemia in chronic hepatitis C patients treated with peginterferon alfa-2a (40KD) plus ribavirin. PMID- 17118126 TI - Pharmacokinetics of ribavirin in patients with hepatitis C virus. AB - AIM: A population pharmacokinetic analysis was performed using plasma concentration data (n = 7025) from 380 patients to examine the relationship between ribavirin dose and its pharmacokinetics. METHODS: Ribavirin pharmacokinetics were described by a three-compartment model with sequential zero order and a first-order absorption processes. Interoccasion variability and food effects were included. RESULTS: Lean body weight (range 41-91 kg) was the only covariate with a clinically significant influence on ribavirin pharmacokinetics, affecting clearance (15.3-23.9 l h(-1)) and the volume of the larger peripheral compartment. CONCLUSION: The model provided a good description of the available data, confirmed by accurate estimates of parameter values and low residual variability (17%). PMID- 17118127 TI - An ocular medication dispensing error. PMID- 17118128 TI - Clinical pharmacology--a sleeping beauty? PMID- 17118130 TI - Proceedings of the Third Annual Conference of the MidSouth Computational Biology and Bioinformatics Society. Introduction. PMID- 17118131 TI - Permutation test for periodicity in short time series data. AB - BACKGROUND: Periodic processes, such as the circadian rhythm, are important factors modulating and coordinating transcription of genes governing key metabolic pathways. Theoretically, even small fluctuations in the orchestration of circadian gene expression patterns among different tissues may result in functional asynchrony, at the organism level, and may contribute to a wide range of pathologic disorders. Identification of circadian expression pattern in time series data is important, but equally challenging. Microarray technology allows estimation of relative expression of thousands of genes at each time point. However, this estimation often lacks precision and microarray experiments are prohibitively expensive, limiting the number of data points in a time series expression profile. The data produced in these experiments carries a high degree of stochastic variation, obscuring the periodic pattern and a limited number of replicates, typically covering not more than two complete periods of oscillation. RESULTS: To address this issue, we have developed a simple, but effective, computational technique for the identification of a periodic pattern in relatively short time series, typical for microarray studies of circadian expression. This test is based on a random permutation of time points in order to estimate non-randomness of a periodogram. The Permutated time, or Pt-test, is able to detect oscillations within a given period in expression profiles dominated by a high degree of stochastic fluctuations or oscillations of different irrelevant frequencies. We have conducted a comprehensive study of circadian expression on a large data set produced at PBRC, representing three different peripheral murine tissues. We have also re-analyzed a number of similar time series data sets produced and published independently by other research groups over the past few years. CONCLUSION: The Permutated time test (Pt-test) is demonstrated to be effective for detection of periodicity in short time series typical for high-density microarray experiments. The software is a set of C++ programs available from the authors on the open source basis. PMID- 17118132 TI - A method for computing the overall statistical significance of a treatment effect among a group of genes. AB - BACKGROUND: In studies that use DNA arrays to assess changes in gene expression, our goal is to evaluate the statistical significance of treatments on sets of genes. Genes can be grouped by a molecular function, a biological process, or a cellular component, e.g., gene ontology (GO) terms. The meaning of an affected GO group is often clearer than interpretations arising from a list of the statistically significant genes. RESULTS: Computer simulations demonstrated that correlations among genes invalidate many statistical methods that are commonly used to assign significance to GO terms. Ignoring these correlations overstates the statistical significance. Meta-analysis methods for combining p-values were modified to adjust for correlation. One of these methods is elaborated in the context of a comparison between two treatments. The form of the correlation adjustment depends upon the alternative hypothesis. CONCLUSION: Reliable corrections for the effect of correlations among genes on the significance level of a GO term can be constructed for an alternative hypothesis where all transcripts in the GO term increase (decrease) in response to treatment. For general alternatives, which allow some transcripts to increase and others to decrease, the bias of naive significance calculations can be greatly decreased although not eliminated. PMID- 17118133 TI - Improving the performance of SVM-RFE to select genes in microarray data. AB - BACKGROUND: Recursive Feature Elimination is a common and well-studied method for reducing the number of attributes used for further analysis or development of prediction models. The effectiveness of the RFE algorithm is generally considered excellent, but the primary obstacle in using it is the amount of computational power required. RESULTS: Here we introduce a variant of RFE which employs ideas from simulated annealing. The goal of the algorithm is to improve the computational performance of recursive feature elimination by eliminating chunks of features at a time with as little effect on the quality of the reduced feature set as possible. The algorithm has been tested on several large gene expression data sets. The RFE algorithm is implemented using a Support Vector Machine to assist in identifying the least useful gene(s) to eliminate. CONCLUSION: The algorithm is simple and efficient and generates a set of attributes that is very similar to the set produced by RFE. PMID- 17118134 TI - Beyond microarrays: find key transcription factors controlling signal transduction pathways. AB - BACKGROUND: Massive gene expression changes in different cellular states measured by microarrays, in fact, reflect just an "echo" of real molecular processes in the cells. Transcription factors constitute a class of the regulatory molecules that typically require posttranscriptional modifications or ligand binding in order to exert their function. Therefore, such important functional changes of transcription factors are not directly visible in the microarray experiments. RESULTS: We developed a novel approach to find key transcription factors that may explain concerted expression changes of specific components of the signal transduction network. The approach aims at revealing evidence of positive feedback loops in the signal transduction circuits through activation of pathway specific transcription factors. We demonstrate that promoters of genes encoding components of many known signal transduction pathways are enriched by binding sites of those transcription factors that are endpoints of the considered pathways. Application of the approach to the microarray gene expression data on TNF-alpha stimulated primary human endothelial cells helped to reveal novel key transcription factors potentially involved in the regulation of the signal transduction pathways of the cells. CONCLUSION: We developed a novel computational approach for revealing key transcription factors by knowledge-based analysis of gene expression data with the help of databases on gene regulatory networks (TRANSFAC and TRANSPATH. The corresponding software and databases are available at http://www.gene-regulation.com. PMID- 17118135 TI - Hidden Markov model variants and their application. AB - Markov statistical methods may make it possible to develop an unsupervised learning process that can automatically identify genomic structure in prokaryotes in a comprehensive way. This approach is based on mutual information, probabilistic measures, hidden Markov models, and other purely statistical inputs. This approach also provides a uniquely common ground for comparative prokaryotic genomics. The approach is an on-going effort by its nature, as a multi-pass learning process, where each round is more informed than the last, and thereby allows a shift to the more powerful methods available for supervised learning at each iteration. It is envisaged that this "bootstrap" learning process will also be useful as a knowledge discovery tool. For such an ab initio prokaryotic gene-finder to work, however, it needs a mechanism to identify critical motif structure, such as those around the start of coding or start of transcription (and then, hopefully more).For eukaryotes, even with better start of-coding identification, parsing of eukaryotic coding regions by the HMM is still limited by the HMM's single gene assumption, as evidenced by the poor performance in alternatively spliced regions. To address these complications an approach is described to expand the states in a eukaryotic gene-predictor HMM, to operate with two layers of DNA parsing. This extension from the single layer gene prediction parse is indicated after preliminary analysis of the C. elegans alt splice statistics. State profiles have made use of a novel hash-interpolating MM (hIMM) method. A new implementation for an HMM-with-Duration is also described, with far-reaching application to gene-structure identification and analysis of channel current blockade data. PMID- 17118136 TI - DNA molecule classification using feature primitives. AB - BACKGROUND: We present a novel strategy for classification of DNA molecules using measurements from an alpha-Hemolysin channel detector. The proposed approach provides excellent classification performance for five different DNA hairpins that differ in only one base-pair. For multi-class DNA classification problems, practitioners usually adopt approaches that use decision trees consisting of binary classifiers. Finding the best tree topology requires exploring all possible tree topologies and is computationally prohibitive. We propose a computational framework based on feature primitives that eliminates the need of a decision tree of binary classifiers. In the first phase, we generate a pool of weak features from nanopore blockade current measurements by using HMM analysis, principal component analysis and various wavelet filters. In the next phase, feature selection is performed using AdaBoost. AdaBoost provides an ensemble of weak learners of various types learned from feature primitives. RESULTS AND CONCLUSION: We show that our technique, despite its inherent simplicity, provides a performance comparable to recent multi-class DNA molecule classification results. Unlike the approach presented by Winters-Hilt et al., where weaker data is dropped to obtain better classification, the proposed approach provides comparable classification accuracy without any need for rejection of weak data. A weakness of this approach, on the other hand, is the very "hands-on" tuning and feature selection that is required to obtain good generalization. Simply put, this method obtains a more informed set of features and provides better results for that reason. The strength of this approach appears to be in its ability to identify strong features, an area where further results are actively being sought. PMID- 17118137 TI - Analysis of gene expression changes in relation to toxicity and tumorigenesis in the livers of Big Blue transgenic rats fed comfrey (Symphytum officinale). AB - BACKGROUND: Comfrey is consumed by humans as a vegetable and a tea, and has been used as an herbal medicine for more than 2000 years. Comfrey, however, is hepatotoxic in livestock and humans and carcinogenic in experimental animals. Our previous study suggested that comfrey induces liver tumors by a genotoxic mechanism and that the pyrrolizidine alkaloids in the plant are responsible for mutation induction and tumor initiation in rat liver. RESULTS: In this study, we identified comfrey-induced gene expression profile in the livers of rats. Groups of 6 male transgenic Big Blue rats were fed a basal diet and a diet containing 8% comfrey roots, a dose that resulted in liver tumors in a previous carcinogenicity bioassay. The animals were treated for 12 weeks and sacrificed one day after the final treatment. We used a rat microarray containing 26,857 genes to perform genome-wide gene expression studies. Dietary comfrey resulted in marked changes in liver gene expression, as well as in significant decreases in the body weight and increases in liver mutant frequency. When a two-fold cutoff value and a P value less than 0.01 were selected, 2,726 genes were identified as differentially expressed in comfrey-fed rats compared to control animals. Among these genes, there were 1,617 genes associated by Ingenuity Pathway Analysis with particular functions, and the differentially expressed genes in comfrey-fed rat livers were involved in metabolism, injury of endothelial cells, and liver injury and abnormalities, including liver fibrosis and cancer development. CONCLUSION: The gene expression profile provides us a better understanding of underlying mechanisms for comfrey-induced hepatic toxicity. Integration of gene expression changes with known pathological changes can be used to formulate a mechanistic scheme for comfrey-induced liver toxicity and tumorigenesis. PMID- 17118138 TI - Improvement in the reproducibility and accuracy of DNA microarray quantification by optimizing hybridization conditions. AB - BACKGROUND: DNA microarrays, which have been increasingly used to monitor mRNA transcripts at a global level, can provide detailed insight into cellular processes involved in response to drugs and toxins. This is leading to new understandings of signaling networks that operate in the cell, and the molecular basis of diseases. Custom printed oligonucleotide arrays have proven to be an effective way to facilitate the applications of DNA microarray technology. A successful microarray experiment, however, involves many steps: well-designed oligonucleotide probes, printing, RNA extraction and labeling, hybridization, and imaging. Optimization is essential to generate reliable microarray data. RESULTS: Hybridization and washing steps are crucial for a successful microarray experiment. By following the hybridization and washing conditions recommended by an oligonucleotide provider, it was found that the expression ratios were compressed greater than expected and data analysis revealed a high degree of non specific binding. A series of experiments was conducted using rat mixed tissue RNA reference material (MTRRM) and other RNA samples to optimize the hybridization and washing conditions. The optimized hybridization and washing conditions greatly reduced the non-specific binding and improved the accuracy of spot intensity measurements. CONCLUSION: The results from the optimized hybridization and washing conditions greatly improved the reproducibility and accuracy of expression ratios. These experiments also suggested the importance of probe designs using better bioinformatics approaches and the need for common reference RNA samples for platform performance evaluation in order to fulfill the potential of DNA microarray technology. PMID- 17118139 TI - Differential gene expression in mouse primary hepatocytes exposed to the peroxisome proliferator-activated receptor alpha agonists. AB - BACKGROUND: Fibrates are a unique hypolipidemic drugs that lower plasma triglyceride and cholesterol levels through their action as peroxisome proliferator-activated receptor alpha (PPARalpha) agonists. The activation of PPARalpha leads to a cascade of events that result in the pharmacological (hypolipidemic) and adverse (carcinogenic) effects in rodent liver. RESULTS: To understand the molecular mechanisms responsible for the pleiotropic effects of PPARalpha agonists, we treated mouse primary hepatocytes with three PPARalpha agonists (bezafibrate, fenofibrate, and WY-14,643) at multiple concentrations (0, 10, 30, and 100 microM) for 24 hours. When primary hepatocytes were exposed to these agents, transactivation of PPARalpha was elevated as measured by luciferase assay. Global gene expression profiles in response to PPARalpha agonists were obtained by microarray analysis. Among differentially expressed genes (DEGs), there were 4, 8, and 21 genes commonly regulated by bezafibrate, fenofibrate, and WY-14,643 treatments across 3 doses, respectively, in a dose-dependent manner. Treatments with 100 muM of bezafibrate, fenofibrate, and WY-14,643 resulted in 151, 149, and 145 genes altered, respectively. Among them, 121 genes were commonly regulated by at least two drugs. Many genes are involved in fatty acid metabolism including oxidative reaction. Some of the gene changes were associated with production of reactive oxygen species, cell proliferation of peroxisomes, and hepatic disorders. In addition, 11 genes related to the development of liver cancer were observed. CONCLUSION: Our results suggest that treatment of PPARalpha agonists results in the production of oxidative stress and increased peroxisome proliferation, thus providing a better understanding of mechanisms underlying PPARalpha agonist-induced hepatic disorders and hepatocarcinomas. PMID- 17118140 TI - An automated method for rapid identification of putative gene family members in plants. AB - BACKGROUND: Gene duplication events have played a significant role in genome evolution, particularly in plants. Exhaustive searches for all members of a known gene family as well as the identification of new gene families has become increasingly important. Subfunctionalization via changes in regulatory sequences following duplication (adaptive selection) appears to be a common mechanism of evolution in plants and can be accompanied by purifying selection on the coding region. Such negative selection can be detected by a bias toward synonymous over nonsynonymous substitutions. However, the process of identifying this bias requires many steps usually employing several different software programs. We have simplified the process and significantly shortened the time required by condensing many steps into a few scripts or programs to rapidly identify putative gene family members beginning with a single query sequence. RESULTS: In this report we 1) describe the software tools (SimESTs, PCAT, and SCAT) developed to automate the gene family identification, 2) demonstrate the validity of the method by correctly identifying 3 of 4 PAL gene family members from Arabidopsis using EST data alone, 3) identify 2 to 6 CAD gene family members from Glycine max (previously unidentified), and 4) identify 2 members of a putative Glycine max gene family previously unidentified in any plant species. CONCLUSION: Gene families in plants, particularly that subset where purifying selection has occurred in the coding region, can be identified quickly and easily by integrating our software tools and commonly available contig assembly and ORF identification programs. PMID- 17118141 TI - A Fourier transformation based method to mine peptide space for antimicrobial activity. AB - BACKGROUND: Naturally occurring antimicrobial peptides are currently being explored as potential candidate peptide drugs. Since antimicrobial peptides are part of the innate immune system of every living organism, it is possible to discover new candidate peptides using the available genomic and proteomic data. High throughput computational techniques could also be used to virtually scan the entire peptide space for discovering out new candidate antimicrobial peptides. RESULT: We have identified a unique indexing method based on biologically distinct characteristic features of known antimicrobial peptides. Analysis of the entries in the antimicrobial peptide databases, based on our indexing method, using Fourier transformation technique revealed a distinct peak in their power spectrum. We have developed a method to mine the genomic and proteomic data, for the presence of peptides with potential antimicrobial activity, by looking for this distinct peak. We also used the Euclidean metric to rank the potential antimicrobial peptides activity. We have parallelized our method so that virtually any given protein space could be data mined, in search of antimicrobial peptides. CONCLUSION: The results show that the Fourier transform based method with the property based coding strategy could be used to scan the peptide space for discovering new potential antimicrobial peptides. PMID- 17118142 TI - Gene expression profiles distinguish the carcinogenic effects of aristolochic acid in target (kidney) and non-target (liver) tissues in rats. AB - BACKGROUND: Aristolochic acid (AA) is the active component of herbal drugs derived from Aristolochia species that have been used for medicinal purposes since antiquity. AA, however, induced nephropathy and urothelial cancer in people and malignant tumors in the kidney and urinary tract of rodents. Although AA is bioactivated in both kidney and liver, it only induces tumors in kidney. To evaluate whether microarray analysis can be used for distinguishing the tissue specific carcinogenicity of AA, we examined gene expression profiles in kidney and liver of rats treated with carcinogenic doses of AA. RESULTS: Microarray analysis was performed using the Rat Genome Survey Microarray and data analysis was carried out within ArrayTrack software. Principal components analysis and hierarchical cluster analysis of the expression profiles showed that samples were grouped together according to the tissues and treatments. The gene expression profiles were significantly altered by AA treatment in both kidney and liver (p < 0.01; fold change > 1.5). Functional analysis with Ingenuity Pathways Analysis showed that there were many more significantly altered genes involved in cancer related pathways in kidney than in liver. Also, analysis with Gene Ontology for Functional Analysis (GOFFA) software indicated that the biological processes related to defense response apoptosis and immune response were significantly altered by AA exposure in kidney, but not in liver. CONCLUSION: Our results suggest that microarray analysis is a useful tool for detecting AA exposure; that analysis of the gene expression profiles can define the differential responses to toxicity and carcinogenicity of AA from kidney and liver; and that significant alteration of genes associated with defense response, apoptosis and immune response in kidney, but not in liver, may be responsible for the tissue-specific toxicity and carcinogenicity of AA. PMID- 17118143 TI - Nanopore Detector based analysis of single-molecule conformational kinetics and binding interactions. AB - BACKGROUND: A Nanopore Detector provides a means to transduce single molecule events into observable channel current changes. Nanopore-based detection can report directly, or indirectly, on single molecule kinetics. The nanopore-based detector can directly measure molecular characteristics in terms of the blockade properties of individual molecules--this is possible due to the kinetic information that is embedded in the blockade measurements, where the adsorption desorption history of the molecule to the surrounding channel, and the configurational changes in the molecule itself, imprint on the ionic flow through the channel. This rich source of information offers prospects for DNA sequencing and single nucleotide polymorphism (SNP) analysis. A nanopore-based detector can also measure molecular characteristics indirectly, by using a reporter molecule that binds to certain molecules, with subsequent distinctive blockade by the bound-molecule complex. RESULTS: It is hypothesized that reaction histories of individual molecules can be observed on model DNA/DNA, DNA/Protein, and Protein/Protein systems. Preliminary results are all consistent with this hypothesis. Nanopore detection capabilities are also described for highly discriminatory biosensing, binding strength characterization, and rapid immunological screening. CONCLUSION: In essence, the heart of chemistry is now accessible to a new, single-molecule, observation method that can track both external molecular binding states, and internal conformation states. PMID- 17118144 TI - Cheminformatics methods for novel nanopore analysis of HIV DNA termini. AB - BACKGROUND: Channel current feature extraction methods, using Hidden Markov Models (HMMs) have been designed for tracking individual-molecule conformational changes. This information is derived from observation of changes in ionic channel current blockade "signal" upon that molecule's interaction with (and occlusion of) a single nanometer-scale channel in a "nanopore detector". In effect, a nanopore detector transduces single molecule events into channel current blockades. HMM analysis tools described are used to help systematically explore DNA dinucleotide flexibility, with particular focus on HIV's highly conserved (and highly flexible/reactive) viral DNA termini. One of the most critical stages in HIV's attack is the binding between viral DNA and the retroviral integrase, which is influenced by the dynamic-coupling induced high flexibility of a CA/TG dinucleotide positioned precisely two base-pairs from the blunt terminus of the duplex viral DNA. This suggests the study of a family of such CA/TG dinucleotide molecules via nanopore measurement and cheminformatics analysis. RESULTS: HMMs are used for level identification on the current blockades, HMM/EM with boosted variance emissions are used for level projection pre-processing, and time-domain FSAs are used to parse the level-projected waveform for kinetic information. The observed state kinetics of the DNA hairpins containing the CA/TG dinucleotide provides clear evidence for HIV's selection of a peculiarly flexible/interactive DNA terminus. PMID- 17118145 TI - GOFFA: gene ontology for functional analysis--a FDA gene ontology tool for analysis of genomic and proteomic data. AB - BACKGROUND: Gene Ontology (GO) characterizes and categorizes the functions of genes and their products according to biological processes, molecular functions and cellular components, facilitating interpretation of data from high-throughput genomics and proteomics technologies. The most effective use of GO information is achieved when its rich and hierarchical complexity is retained and the information is distilled to the biological functions that are most germane to the phenomenon being investigated. RESULTS: Here we present a FDA GO tool named Gene Ontology for Functional Analysis (GOFFA). GOFFA first ranks GO terms in the order of prevalence for a list of selected genes or proteins, and then it allows the user to interactively select GO terms according to their significance and specific biological complexity within the hierarchical structure. GOFFA provides five interactive functions (Tree view, Terms View, Genes View, GO Path and GO TreePrune) to analyze the GO data. Among the five functions, GO Path and GO TreePrune are unique. The GO Path simultaneously displays the ranks that order GOFFA Tree Paths based on statistical analysis. The GO TreePrune provides a visual display of a reduced GO term set based on a user's statistical cut-offs. Therefore, the GOFFA visual display can provide an intuitive depiction of the most likely relevant biological functions. CONCLUSION: With GOFFA, the user can dynamically interact with the GO data to interpret gene expression results in the context of biological plausibility, which can lead to new discoveries or identify new hypotheses. AVAILABILITY: GOFFA is available through ArrayTrack softwarehttp://edkb.fda.gov/webstart/arraytrack/. PMID- 17118146 TI - A scalable machine-learning approach to recognize chemical names within large text databases. AB - MOTIVATION: The use or study of chemical compounds permeates almost every scientific field and in each of them, the amount of textual information is growing rapidly. There is a need to accurately identify chemical names within text for a number of informatics efforts such as database curation, report summarization, tagging of named entities and keywords, or the development/curation of reference databases. RESULTS: A first-order Markov Model (MM) was evaluated for its ability to distinguish chemical names from words, yielding approximately 93% recall in recognizing chemical terms and approximately 99% precision in rejecting non-chemical terms on smaller test sets. However, because total false-positive events increase with the number of words analyzed, the scalability of name recognition was measured by processing 13.1 million MEDLINE records. The method yielded precision ranges from 54.7% to 100%, depending upon the cutoff score used, averaging 82.7% for approximately 1.05 million putative chemical terms extracted. Extracted chemical terms were analyzed to estimate the number of spelling variants per term, which correlated with the total number of times the chemical name appeared in MEDLINE. This variability in term construction was found to affect both information retrieval and term mapping when using PubMed and Ovid. PMID- 17118147 TI - Support vector machine implementations for classification & clustering. AB - BACKGROUND: We describe Support Vector Machine (SVM) applications to classification and clustering of channel current data. SVMs are variational calculus based methods that are constrained to have structural risk minimization (SRM), i.e., they provide noise tolerant solutions for pattern recognition. The SVM approach encapsulates a significant amount of model-fitting information in the choice of its kernel. In work thus far, novel, information-theoretic, kernels have been successfully employed for notably better performance over standard kernels. Currently there are two approaches for implementing multiclass SVMs. One is called external multi-class that arranges several binary classifiers as a decision tree such that they perform a single-class decision making function, with each leaf corresponding to a unique class. The second approach, namely internal-multiclass, involves solving a single optimization problem corresponding to the entire data set (with multiple hyperplanes). RESULTS: Each SVM approach encapsulates a significant amount of model-fitting information in its choice of kernel. In work thus far, novel, information-theoretic, kernels were successfully employed for notably better performance over standard kernels. Two SVM approaches to multiclass discrimination are described: (1) internal multiclass (with a single optimization), and (2) external multiclass (using an optimized decision tree). We describe benefits of the internal-SVM approach, along with further refinements to the internal-multiclass SVM algorithms that offer significant improvement in training time without sacrificing accuracy. In situations where the data isn't clearly separable, making for poor discrimination, signal clustering is used to provide robust and useful information--to this end, novel, SVM-based clustering methods are also described. As with the classification, there are Internal and External SVM Clustering algorithms, both of which are briefly described. PMID- 17118148 TI - Metric for measuring the effectiveness of clustering of DNA microarray expression. AB - BACKGROUND: The recent advancement of microarray technology with lower noise and better affordability makes it possible to determine expression of several thousand genes simultaneously. The differentially expressed genes are filtered first and then clustered based on the expression profiles of the genes. A large number of clustering algorithms and distance measuring matrices are proposed in the literature. The popular ones among them include hierarchal clustering and k means clustering. These algorithms have often used the Euclidian distance or Pearson correlation distance. The biologists or the practitioners are often confused as to which algorithm to use since there is no clear winner among algorithms or among distance measuring metrics. Several validation indices have been proposed in the literature and these are based directly or indirectly on distances; hence a method that uses any of these indices does not relate to any biological features such as biological processes or molecular functions. RESULTS: In this paper we have proposed a metric to measure the effectiveness of clustering algorithms of genes by computing inter-cluster cohesiveness and as well as the intra-cluster separation with respect to biological features such as biological processes or molecular functions. We have applied this metric to the clusters on the data set that we have created as part of a larger study to determine the cancer suppressive mechanism of a class of chemicals called retinoids.We have considered hierarchal and k-means clustering with Euclidian and Pearson correlation distances. Our results show that genes of similar expression profiles are more likely to be closely related to biological processes than they are to molecular functions. The findings have been supported by many works in the area of gene clustering. CONCLUSION: The best clustering algorithm of genes must achieve cohesiveness within a cluster with respect to some biological features, and as well as maximum separation between clusters in terms of the distribution of genes of a behavioral group across clusters. We claim that our proposed metric is novel in this respect and that it provides a measure of both inter and intra cluster cohesiveness. Best of all, computation of the proposed metric is easy and it provides a single quantitative value, which makes comparison of different algorithms easier. The maximum cluster cohesiveness and the maximum intra-cluster separation are indicated by the metric when its value is 0.We have demonstrated the metric by applying it to a data set with gene behavioral groupings such as biological process and molecular functions. The metric can be easily extended to other features of a gene such as DNA binding sites and protein-protein interactions of the gene product, special features of the intron-exon structure, promoter characteristics, etc. The metric can also be used in other domains that use two different parametric spaces; one for clustering and the other one for measuring the effectiveness. PMID- 17118149 TI - RiboaptDB: a comprehensive database of ribozymes and aptamers. AB - BACKGROUND: Catalytic RNA molecules are called ribozymes. The aptamers are DNA or RNA molecules that have been selected from vast populations of random sequences, through a combinatorial approach known as SELEX. The selected oligo-nucleotide sequences (~200 bp in length) have the ability to recognize broad range of specific ligands by forming binding pockets. These novel aptamer sequences can bind to nucleic acids, proteins or small organic and inorganic chemical compounds and have many potential uses in medicine and technology. RESULTS: The comprehensive sequence information on aptamers and ribozymes that have been generated by in vitro selection methods are included in this RiboaptDB database. Such types of unnatural data generated by in vitro methods are not available in the public 'natural' sequence databases such as GenBank and EMBL. The amount of sequence data generated by in vitro selection experiments has been accumulating exponentially. There are 370 artificial ribozyme sequences and 3842 aptamer sequences in the total 4212 sequences from 423 citations in this RiboaptDB. We included general search feature, and individual feature wise search, user submission form for new data through online and also local BLAST search. CONCLUSION: This database, besides serving as a storehouse of sequences that may have diagnostic or therapeutic utility in medicine, provides valuable information for computational and theoretical biologists. The RiboaptDB is extremely useful for garnering information about in vitro selection experiments as a whole and for better understanding the distribution of functional nucleic acids in sequence space. The database is updated regularly and is publicly available at http://mfgn.usm.edu/ebl/riboapt/. PMID- 17118150 TI - EGenBio: a data management system for evolutionary genomics and biodiversity. AB - BACKGROUND: Evolutionary genomics requires management and filtering of large numbers of diverse genomic sequences for accurate analysis and inference on evolutionary processes of genomic and functional change. We developed Evolutionary Genomics and Biodiversity (EGenBio; http://egenbio.lsu.edu) to begin to address this. DESCRIPTION: EGenBio is a system for manipulation and filtering of large numbers of sequences, integrating curated sequence alignments and phylogenetic trees, managing evolutionary analyses, and visualizing their output. EGenBio is organized into three conceptual divisions, Evolution, Genomics, and Biodiversity. The Genomics division includes tools for selecting pre-aligned sequences from different genes and species, and for modifying and filtering these alignments for further analysis. Species searches are handled through queries that can be modified based on a tree-based navigation system and saved. The Biodiversity division contains tools for analyzing individual sequences or sequence alignments, whereas the Evolution division contains tools involving phylogenetic trees. Alignments are annotated with analytical results and modification history using our PRAED format. A miscellaneous Tools section and Help framework are also available. EGenBio was developed around our comparative genomic research and a prototype database of mtDNA genomes. It utilizes MySQL relational databases and dynamic page generation, and calls numerous custom programs. CONCLUSION: EGenBio was designed to serve as a platform for tools and resources to ease combined analysis in evolution, genomics, and biodiversity. PMID- 17118151 TI - Independent component analysis-motivated approach to classificatory decomposition of cortical evoked potentials. AB - BACKGROUND: Independent Component Analysis (ICA) proves to be useful in the analysis of neural activity, as it allows for identification of distinct sources of activity. Applied to measurements registered in a controlled setting and under exposure to an external stimulus, it can facilitate analysis of the impact of the stimulus on those sources. The link between the stimulus and a given source can be verified by a classifier that is able to "predict" the condition a given signal was registered under, solely based on the components. However, the ICA's assumption about statistical independence of sources is often unrealistic and turns out to be insufficient to build an accurate classifier. Therefore, we propose to utilize a novel method, based on hybridization of ICA, multi-objective evolutionary algorithms (MOEA), and rough sets (RS), that attempts to improve the effectiveness of signal decomposition techniques by providing them with "classification-awareness." RESULTS: The preliminary results described here are very promising and further investigation of other MOEAs and/or RS-based classification accuracy measures should be pursued. Even a quick visual analysis of those results can provide an interesting insight into the problem of neural activity analysis. CONCLUSION: We present a methodology of classificatory decomposition of signals. One of the main advantages of our approach is the fact that rather than solely relying on often unrealistic assumptions about statistical independence of sources, components are generated in the light of a underlying classification problem itself. PMID- 17118152 TI - Microarray analysis distinguishes differential gene expression patterns from large and small colony Thymidine kinase mutants of L5178Y mouse lymphoma cells. AB - BACKGROUND: The Thymidine kinase (Tk) mutants generated from the widely used L5178Y mouse lymphoma assay fall into two categories, small colony and large colony. Cells from the large colonies grow at a normal rate while cells from the small colonies grow slower than normal. The relative proportion of large and small colonies after mutagen treatment is associated with a mutagen's ability to induce point mutations and/or chromosomal mutations. The molecular distinction between large and small colony mutants, however, is not clear. RESULTS: To gain insights into the underlying mechanisms responsible for the mutant colony phenotype, microarray gene expression analysis was carried out on 4 small and 4 large colony Tk mutant samples. NCTR-fabricated long-oligonucleotide microarrays of 20,000 mouse genes were used in a two-color reference design experiment. The data were analyzed within ArrayTrack software that was developed at the NCTR. Principal component analysis and hierarchical clustering of the gene expression profiles showed that the samples were clearly separated into two groups based on their colony size phenotypes. The Welch T-test was used for determining significant changes in gene expression between the large and small colony groups and 90 genes whose expression was significantly altered were identified (p < 0.01; fold change > 1.5). Using Ingenuity Pathways Analysis (IPA), 50 out of the 90 significant genes were found in the IPA database and mapped to four networks associated with cell growth. Eleven percent of the 90 significant genes were located on chromosome 11 where the Tk gene resides while only 5.6% of the genes on the microarrays mapped to chromosome 11. All of the chromosome 11 significant genes were expressed at a higher level in the small colony mutants compared to the large colony mutants. Also, most of the significant genes located on chromosome 11 were disproportionally concentrated on the distal end of chromosome 11 where the Tk mutations occurred. CONCLUSION: The results indicate that microarray analysis can define cellular phenotypes and identify genes that are related to the colony size phenotypes. The findings suggest that genes in the DNA segment altered by the Tk mutations were significantly up-regulated in the small colony mutants, but not in the large colony mutants, leading to differential expression of a set of growth regulation genes that are related to cell apoptosis and other cellular functions related to the restriction of cell growth. PMID- 17118154 TI - Mathematical modeling of intracellular signaling pathways. AB - Dynamic modeling and simulation of signal transduction pathways is an important topic in systems biology and is obtaining growing attention from researchers with experimental or theoretical background. Here we review attempts to analyze and model specific signaling systems. We review the structure of recurrent building blocks of signaling pathways and their integration into more comprehensive models, which enables the understanding of complex cellular processes. The variety of mechanisms found and modeling techniques used are illustrated with models of different signaling pathways. Focusing on the close interplay between experimental investigation of pathways and the mathematical representations of cellular dynamics, we discuss challenges and perspectives that emerge in studies of signaling systems. PMID- 17118155 TI - Model storage, exchange and integration. AB - The field of Computational Systems Neurobiology is maturing quickly. If one wants it to fulfil its central role in the new Integrative Neurobiology, the reuse of quantitative models needs to be facilitated. The community has to develop standards and guidelines in order to maximise the diffusion of its scientific production, but also to render it more trustworthy. In the recent years, various projects tackled the problems of the syntax and semantics of quantitative models. More recently the international initiative BioModels.net launched three projects: (1) MIRIAM is a standard to curate and annotate models, in order to facilitate their reuse. (2) The Systems Biology Ontology is a set of controlled vocabularies aimed to be used in conjunction with models, in order to characterise their components. (3) BioModels Database is a resource that allows biologists to store, search and retrieve published mathematical models of biological interests. We expect that those resources, together with the use of formal languages such as SBML, will support the fruitful exchange and reuse of quantitative models. PMID- 17118157 TI - Rules of engagement promote polarity in RNA trafficking. AB - Many cell biological pathways exhibit overall polarity (net movement of molecules in one direction) even though individual molecular interactions in the pathway are freely reversible. The A2 RNA trafficking pathway exhibits polarity in moving specific RNA molecules from the nucleus to localization sites in the myelin compartment of oligodendrocytes or dendritic spines in neurons. The A2 pathway is mediated by a ubiquitously expressed trans-acting trafficking factor (hnRNP A2) that interacts with a specific 11 nucleotide cis-acting trafficking sequence termed the A2 response element (A2RE) found in several localized RNAs. Five different molecular partners for hnRNP A2 have been identified in the A2 pathway: hnRNP A2 itself, transportin, A2RE RNA, TOG (tumor overexpressed gene) and hnRNP E1, each playing a key role in one particular step of the A2 pathway. Sequential interactions of hnRNP A2 with different molecular partners at each step mediate directed movement of trafficking intermediates along the pathway. Specific "rules of engagement" (both and, either or, only if) govern sequential interactions of hnRNP A2 with each of its molecular partners. Rules of engagement are defined experimentally using three component binding assays to measure differential binding of hnRNP A2 to one partner in the presence of each of the other partners in the pathway. Here we describe rules of engagement for hnRNP A2 binding to each of its molecular partners and discuss how these rules of engagement promote polarity in the A2 RNA trafficking pathway. For molecules with multiple binding partners, specific rules of engagement govern different molecular interactions. Rules of engagement are ultimately determined by structural relationships between binding sites on individual molecules. In the A2 RNA trafficking pathway rules of engagement governing interactions of hnRNP A2 with different binding partners provide the basis for polarity of movement of intermediates along the pathway. PMID- 17118156 TI - The modular systems biology approach to investigate the control of apoptosis in Alzheimer's disease neurodegeneration. AB - Apoptosis is a programmed cell death that plays a critical role during the development of the nervous system and in many chronic neurodegenerative diseases, including Alzheimer's disease (AD). This pathology, characterized by a progressive degeneration of cholinergic function resulting in a remarkable cognitive decline, is the most common form of dementia with high social and economic impact. Current therapies of AD are only symptomatic, therefore the need to elucidate the mechanisms underlying the onset and progression of the disease is surely needed in order to develop effective pharmacological therapies. Because of its pivotal role in neuronal cell death, apoptosis has been considered one of the most appealing therapeutic targets, however, due to the complexity of the molecular mechanisms involving the various triggering events and the many signaling cascades leading to cell death, a comprehensive understanding of this process is still lacking. Modular systems biology is a very effective strategy in organizing information about complex biological processes and deriving modular and mathematical models that greatly simplify the identification of key steps of a given process. This review aims at describing the main steps underlying the strategy of modular systems biology and briefly summarizes how this approach has been successfully applied for cell cycle studies. Moreover, after giving an overview of the many molecular mechanisms underlying apoptosis in AD, we present both a modular and a molecular model of neuronal apoptosis that suggest new insights on neuroprotection for this disease. PMID- 17118158 TI - Synaptic proteins as multi-sensor devices of neurotransmission. AB - Neuronal communication is tightly regulated in time and space. Following neuronal activation, an electrical signal triggers neurotransmitter (NT) release at the active zone. The process starts by the signal reaching the synapse followed by a fusion of the synaptic vesicle (SV) and diffusion of the released NT in the synaptic cleft. The NT then binds to the appropriate receptor and induces a membrane potential change at the target cell membrane. The entire process is controlled by a fairly small set of synaptic proteins, collectively called SYCONs. The biochemical features of SYCONs underlie the properties of NT release. SYCONs are characterized by their ability to detect and respond to changes in environmental signals. For example, consider synaptotagmin I (Syt1), a prototype of a protein family with over 20 gene and variants in mammals. Syt1 is a specific example of a multi-sensor device with a large repertoire of discrete states. Several of these states are stimulated by a local concentration of signaling molecules such as Ca2+. The ability of this protein to sense signaling molecules and to adopt multiple biochemical states is shared by other SYCONs such as the synapsins (Syns). Specific biochemical states of Syns determine the accessibility of SV for NT release. Each of these states is defined by a specific alternative spliced variant with a unique profile of phosphorylation modified sites. The plasticity of the synapse is a direct reflection of SYCON's multiple biochemical states. State transitions occurs in a wide range of time scales, and therefore these molecules need to cope with events that last milliseconds (i.e., exocytosis in fast responding synapses) and with events that can carry on for many minutes (i.e., organization of SV pools). We suggest that SYCONs are optimized throughout evolution as multi-sensor devices. A full repertoire of the switches leading to alternation of protein states and a detailed characterization of protein-protein network within the synapse is critical for the development of a dynamic model of synaptic transmission. PMID- 17118159 TI - Non coding RNA and brain. AB - Small non coding RNAs are a group of very different RNA molecules, present in virtually all cells, with a wide spectrum of regulatory functions which include RNA modification and regulation of protein synthesis. They have been isolated and characterized in all organisms and tissues, from Archaeobacteria to mammals. In mammalian brain there are a number of these small molecules, which are involved in neuronal differentiation as well as, possibly, in learning and memory. In this manuscript, we analyze the present knowledge about the function of the most important groups of small non-coding RNA present in brain: small nucleolar RNAs, small cytoplasmic RNAs, and microRNAs. The last ones, in particular, appear to be critical for dictating neuronal cell identity during development and to play an important role in neurite growth, synaptic development and neuronal plasticity. PMID- 17118160 TI - Parameter estimate of signal transduction pathways. AB - BACKGROUND: The "inverse" problem is related to the determination of unknown causes on the bases of the observation of their effects. This is the opposite of the corresponding "direct" problem, which relates to the prediction of the effects generated by a complete description of some agencies. The solution of an inverse problem entails the construction of a mathematical model and takes the moves from a number of experimental data. In this respect, inverse problems are often ill-conditioned as the amount of experimental conditions available are often insufficient to unambiguously solve the mathematical model. Several approaches to solving inverse problems are possible, both computational and experimental, some of which are mentioned in this article. In this work, we will describe in details the attempt to solve an inverse problem which arose in the study of an intracellular signaling pathway. RESULTS: Using the Genetic Algorithm to find the sub-optimal solution to the optimization problem, we have estimated a set of unknown parameters describing a kinetic model of a signaling pathway in the neuronal cell. The model is composed of mass action ordinary differential equations, where the kinetic parameters describe protein-protein interactions, protein synthesis and degradation. The algorithm has been implemented on a parallel platform. Several potential solutions of the problem have been computed, each solution being a set of model parameters. A sub-set of parameters has been selected on the basis on their small coefficient of variation across the ensemble of solutions. CONCLUSION: Despite the lack of sufficiently reliable and homogeneous experimental data, the genetic algorithm approach has allowed to estimate the approximate value of a number of model parameters in a kinetic model of a signaling pathway: these parameters have been assessed to be relevant for the reproduction of the available experimental data. PMID- 17118161 TI - Integration of enzyme kinetic models and isotopomer distribution analysis for studies of in situ cell operation. AB - A current trend in neuroscience research is the use of stable isotope tracers in order to address metabolic processes in vivo. The tracers produce a huge number of metabolite forms that differ according to the number and position of labeled isotopes in the carbon skeleton (isotopomers) and such a large variety makes the analysis of isotopomer data highly complex. On the other hand, this multiplicity of forms does provide sufficient information to address cell operation in vivo. By the end of last millennium, a number of tools have been developed for estimation of metabolic flux profile from any possible isotopomer distribution data. However, although well elaborated, these tools were limited to steady state analysis, and the obtained set of fluxes remained disconnected from their biochemical context. In this review we focus on a new numerical analytical approach that integrates kinetic and metabolic flux analysis. The related computational algorithm estimates the dynamic flux based on the time-dependent distribution of all possible isotopomers of metabolic pathway intermediates that are generated from a labeled substrate. The new algorithm connects specific tracer data with enzyme kinetic characteristics, thereby extending the amount of data available for analysis: it uses enzyme kinetic data to estimate the flux profile, and vice versa, for the kinetic analysis it uses in vivo tracer data to reveal the biochemical basis of the estimated metabolic fluxes. PMID- 17118162 TI - Computational framework for the prediction of transcription factor binding sites by multiple data integration. AB - Control of gene expression is essential to the establishment and maintenance of all cell types, and its dysregulation is involved in pathogenesis of several diseases. Accurate computational predictions of transcription factor regulation may thus help in understanding complex diseases, including mental disorders in which dysregulation of neural gene expression is thought to play a key role. However, biological mechanisms underlying the regulation of gene expression are not completely understood, and predictions via bioinformatics tools are typically poorly specific. We developed a bioinformatics workflow for the prediction of transcription factor binding sites from several independent datasets. We show the advantages of integrating information based on evolutionary conservation and gene expression, when tackling the problem of binding site prediction. Consistent results were obtained on a large simulated dataset consisting of 13050 in silico promoter sequences, on a set of 161 human gene promoters for which binding sites are known, and on a smaller set of promoters of Myc target genes. Our computational framework for binding site prediction can integrate multiple sources of data, and its performance was tested on different datasets. Our results show that integrating information from multiple data sources, such as genomic sequence of genes' promoters, conservation over multiple species, and gene expression data, indeed improves the accuracy of computational predictions. PMID- 17118163 TI - Mathematical modelling and numerical simulation of the morphological development of neurons. AB - BACKGROUND: The morphological development of neurons is a very complex process involving both genetic and environmental components. Mathematical modelling and numerical simulation are valuable tools in helping us unravel particular aspects of how individual neurons grow their characteristic morphologies and eventually form appropriate networks with each other. METHODS: A variety of mathematical models that consider (1) neurite initiation (2) neurite elongation (3) axon pathfinding, and (4) neurite branching and dendritic shape formation are reviewed. The different mathematical techniques employed are also described. RESULTS: Some comparison of modelling results with experimental data is made. A critique of different modelling techniques is given, leading to a proposal for a unified modelling environment for models of neuronal development. CONCLUSION: A unified mathematical and numerical simulation framework should lead to an expansion of work on models of neuronal development, as has occurred with compartmental models of neuronal electrical activity. PMID- 17118164 TI - Basic and applied biology of the primate reproductive tract--a symposium in honor of the career of Dr Robert M Brenner. PMID- 17118165 TI - The role of HLA-G in human pregnancy. AB - Pregnancy in mammals featuring hemochorial placentation introduces a major conflict with the mother's immune system, which is dedicated to repelling invaders bearing foreign DNA and RNA. Numerous and highly sophisticated strategies for preventing mothers from rejecting their genetically different fetus(es) have now been identified. These involve production of novel soluble and membrane-bound molecules by uterine and placental cells. In humans, the placenta derived molecules include glycoproteins derived from the HLA class Ib gene, HLA G. Isoforms of HLA-G saturate the maternal-fetal interface and circulate in mothers throughout pregnancy. Uteroplacental immune privilege for the fetus and its associated tissues is believed to result when immune cells encounter HLA-G. Unequivocally demonstration of this concept requires experiments in animal models. Both the monkey and the baboon express molecules that are similar but not identical to HLA-G, and may comprise suitable animal models for establishing a central role for these proteins in pregnancy. PMID- 17118166 TI - Basic and applied biology of the primate reproductive tract--a symposium in honor of the career of Dr Robert M Brenner: introduction and overview. PMID- 17118167 TI - Cellular and molecular regulation of the primate endometrium: a perspective. AB - This contribution will trace some of the many seminal studies on the female uterus (endometrium) over the centuries and conclude with a description of some current research initiatives in our laboratory. Numerous contributions from many investigators over the years have contributed to our current understanding of endometrial function. The historical section of this chapter is intended to be a brief overall description of some of these efforts and not exhaustive. Additional information can be found in the review articles and books cited herein. PMID- 17118168 TI - Application of functional genomics to primate endometrium: insights into biological processes. AB - Endometrium is a dynamic tissue that responds on a cyclic basis to circulating levels of the ovarian-derived steroid hormones, estradiol and progesterone. Functional genomics has enabled a global approach to understanding gene regulation in whole endometrial tissue in the setting of a changing hormonal milieu. The proliferative phase of the cycle, under the influence of estradiol, has a preponderance of genes involved in DNA synthesis and cell cycle regulation. Interestingly, genes encoding ion channels and cell adhesion, as well as angiogenic factors, are also highly regulated in this phase of the cycle. After the LH surge, different gene expression profiles are uniquely observed in the early secretory, mid-secretory (window of implantation), and late secretory phases. The early secretory phase is notable for up-regulation of multiple genes and gene families involved in cellular metabolism, steroid hormone metabolism, as well as some secreted glycoproteins. The mid-secretory phase is characterized by multiple biological processes, including up-regulation of genes encoding secreted glycoproteins, immune response genes with a focus on innate immunity, and genes involved in detoxification mechanisms. In the late secretory phase, as the tissue prepares for desquamation, there is a marked up-regulation of an inflammatory response, along with matrix degrading enzymes, and genes involved in hemostasis, among others. This monograph reviews hormonal regulation of gene expression in this tissue and the molecular events occurring therein throughout the cycle derived from functional genomics analysis. It also highlights challenges encountered in using human endometrial tissue in translational research in this context. PMID- 17118169 TI - Regulation of human endometrial function: mechanisms relevant to uterine bleeding. AB - This review focuses on the complex events that occur in the endometrium after progesterone is withdrawn (or blocked) and menstrual bleeding ensues. A detailed understanding of these local mechanisms will enhance our knowledge of disturbed endometrial/uterine function--including problems with excessively heavy menstrual bleeding, endometriosis and breakthrough bleeding with progestin only contraception. The development of novel strategies to manage these clinically significant problems depends on such new understanding as does the development of new contraceptives which avoid the endometrial side effect of breakthrough bleeding. PMID- 17118170 TI - A critical period of progesterone withdrawal precedes menstruation in macaques. AB - Macaques are menstruating nonhuman primates that provide important animal models for studies of hormonal regulation in the uterus. In women and macaques the decline of progesterone (P) at the end of the cycle triggers endometrial expression of a variety of matrix metalloproteinase (MMP) enzymes that participate in tissue breakdown and menstrual sloughing. To determine the minimal duration of P withdrawal required to induce menses, we assessed the effects of adding P back at various time points after P withdrawal on both frank bleeding patterns and endometrial MMP expression. Artificial menstrual cycles were induced by treating the animals sequentially with implants releasing estradiol (E2) and progesterone (P). To assess bleeding patterns, P implants were removed at the end of a cycle and then added back at 12, 24, 30, 36, 40, 48, 60, or 72 hours (h) after the initial P withdrawal. Observational analysis of frank bleeding patterns showed that P replacement at 12 and 24 h blocked menses, replacement at 36 h reduced menses but replacement after 36 h failed to block menses. These data indicate that in macaques, a critical period of P withdrawal exists and lasts approximately 36 h. In other similarly cycled animals, we withdrew P and then added P back either during (12-24 h) or after (48 h) the critical period, removed the uterus 24 h after P add back and evaluated endometrial MMP expression. Immunocytochemistry showed that replacement of P during the critical period suppressed MMP-1, -2 and -3 expression along with menses, but replacement of P at 48 h, which failed to suppress mense, suppressed MMP-1 and MMP-3 but did not block MMP-2. We concluded that upregulation of MMPs is essential to menses induction, but that after the critical period, menses will occur even if some MMPs are experimentally blocked. PMID- 17118171 TI - A baboon model for endometriosis: implications for fertility. AB - Endometriosis is one of the most common causes of chronic pelvic pain and infertility in women in the reproductive age group. Although the existence of this disease has been known for over 100 years our current knowledge of its pathogenesis and the pathophysiology of its related infertility remains unclear. Several reasons contribute to our lack of knowledge, the most critical being the difficulty in carrying out objective long-term studies in women. Thus, we and others have developed a model of this disease in the non-human primate, the baboon (Papio anubis). Intraperitoneal inoculation of autologous menstrual endometrium results in the development of endometriotic lesions with gross morphological characteristics similar to those seen in the human. Multiple factors have been implicated in endometriosis-associated infertility. We have described aberrant levels of factors involved in multiple pathways important in the establishment of pregnancy, in the endometrium of baboons induced with endometriosis. Specifically, we have observed dysregulation of proteins involved in invasion, angiogenesis, methylation, cell growth, immunomodulation, and steroid hormone action. These data suggest that, in an induced model of endometriosis in the baboon, an increased angiogenic capacity, decreased apoptotic potential, progesterone resistance, estrogen hyper-responsiveness, and an inability to respond appropriately to embryonic signals contribute to the reduced fecundity associated with this disease. PMID- 17118172 TI - Role of nonhuman primate models in the discovery and clinical development of selective progesterone receptor modulators (SPRMs). AB - Selective progesterone receptor modulators (SPRMs) represent a new class of progesterone receptor ligands that exert clinically relevant tissue-selective progesterone agonist, antagonist, partial, or mixed agonist/antagonist effects on various progesterone target tissues in an in vivo situation depending on the biological action studied. The SPRM asoprisnil is being studied in women with symptomatic uterine leiomyomata and endometriosis. Asoprisnil shows a high degree of uterine selectivity as compared to effects on ovulation or ovarian hormone secretion in humans. It induces amenorrhea and decreases leiomyoma volume in a dose-dependent manner in the presence of follicular phase estrogen concentrations. It also has endometrial antiproliferative effects. In pregnant animals, the myometrial, i.e. labor-inducing, effects of asoprisnil are blunted or absent. Studies in non-human primates played a key role during the preclinical development of selective progesterone receptor modulators. These studies provided the first evidence of uterus-selective effects of asoprisnil and structurally related compounds, and the rationale for clinical development of asoprisnil. PMID- 17118173 TI - Estrogen receptor-alpha (ER-alpha) and defects in uterine receptivity in women. AB - Endometriosis is a disorder that affects 5% of the normal population but is present in up to 40% of women with pelvic pain and/or infertility. Recent evidence suggests that the endometrium of women with endometriosis exhibits progesterone insensitivity. One endometrial protein that fluctuates in response to progesterone is the estrogen receptor-alpha (ER alpha), being down-regulated at the time of peak progesterone secretion during the window of implantation. Here we demonstrate that the biomarker of uterine receptivity, beta 3 integrin subunit, is reduced or absent in some women with endometriosis and that such defects are accompanied by inappropriate over-expression of ER alpha during the mid-secretory phase. Using a well-differentiated endometrial cell line we showed that the beta 3 integrin protein is negatively regulated by estrogen and positively regulated by epidermal growth factor (EGF). By competing against estrogen with various selective estrogen receptor modulators (SERMs) and estrogen receptor agonists and antagonists, inhibition of expression of the beta 3 integrin by estrogen can be mitigated. In conclusion, we hypothesize that certain types of uterine receptivity defects may be caused by the loss of appropriate ER alpha down-regulation in the mid-secretory phase, leading to defects in uterine receptivity. Such changes might be effectively treated by timely administration of the appropriate anti-estrogens to artificially block ER alpha and restore normal patterns of gene expression. Such treatments will require further clinical studies. PMID- 17118174 TI - Occurrence of clinical mastitis in primiparous Estonian dairy cows in different housing conditions. AB - BACKGROUND: Objectives of the study were to document the impact of some management factors on the occurrence of clinical mastitis in primiparous dairy cows and to identify common udder pathogens of clinical mastitis in freshly calved heifers and multiparous cows on the day of calving. METHODS: A one-year study was conducted during 2004 and 2005 in 11 selected Estonian dairy herds. Data consisted of 68 heifers with clinical mastitis and 995 heifers without clinical mastitis on the day of calving. Multivariable logistic regression with a random herd effect was used to investigate any association between housing system or the time interval from movement of heifers to the calving facility and day of calving on occurrence of clinical mastitis. Milk samples for bacteriological analysis were collected from affected heifers and multiparous cows on the day of calving RESULTS: Clinical mastitis occurrence in the study population of freshly calved heifers equalled 6.1 %. Housing system was not a significant risk factor for clinical mastitis of freshly calved heifers. Moving heifers to the cowbarn less than two weeks before calving in tiestall farms increased risk (OR = 5.9 p = 0.001) for clinical mastitis at parturition. The most frequently isolated udder pathogens among heifers were Escherichia coli (22.1%), Streptococcus uberis (19.1%) and coagulase-negative staphylococci (8.8%). In comparison, the main pathogen in multiparous cows with clinical mastitis at parturition was Staphylococcus aureus (11.2%). CONCLUSION: Moving heifers to the calving facilities too late in tiestall farms increased risk for clinical mastitis at parturition. The isolated udder pathogens did not differ significantly in tiestall farms compared to freestall farms in heifers, but differences were found between heifers and multiparous cows at parturition. PMID- 17118175 TI - Postoperative infections after oesophageal resections: the role of blood transfusions. AB - BACKGROUND: Perioperative blood transfusion carries numerous potential risks concerning the transmission of infective diseases and immunodepression that can facilitate the occurrence of postoperative infectious complications. Explanation of connections between perioperative blood transfusion and postoperative septic complication worldwide is not well documented. Many studies have described a correlation between perioperative blood transfusions and postoperative infections. On the contrary, other studies indicate that factors influencing the need for blood transfusions during surgery have a greater bearing than blood transfusion per se on the occurrence of postoperative complications. PATIENTS AND METHODS: A prospective study was conducted in our Department on 110 consecutive patients undergoing oesophageal resection for primary cancer, in order to evaluate the incidence of postoperative infections related to perioperative allogenic blood transfusions. For each patient we preoperatively recorded in a computerized data-base several known risk-factors for postoperative infections; in detail we registered the administration of allogenic perioperative blood transfusions (period of administration, number of packages administered). RESULTS: Among the enrolled 110 patients, 53 (48%) received perioperative blood transfusions: in this group postoperative infections (overall infective complications) occurred in 27 patients. After a multivariate analysis we observed that perioperative blood transfusions significantly affected as an independent variable the development of wound infections (p = 0.02). CONCLUSION: Blood transfusions independently affected the incidence of wound infections in patients who underwent oesophageal resection for primary cancer. PMID- 17118176 TI - Space-time clustering of childhood malaria at the household level: a dynamic cohort in a Mali village. AB - BACKGROUND: Spatial and temporal heterogeneities in the risk of malaria have led the WHO to recommend fine-scale stratification of the epidemiological situation, making it possible to set up actions and clinical or basic researches targeting high-risk zones. Before initiating such studies it is necessary to define local patterns of malaria transmission and infection (in time and in space) in order to facilitate selection of the appropriate study population and the intervention allocation. The aim of this study was to identify, spatially and temporally, high risk zones of malaria, at the household level (resolution of 1 to 3 m). METHODS: This study took place in a Malian village with hyperendemic seasonal transmission as part of Mali-Tulane Tropical Medicine Research Center (NIAID/NIH). The study design was a dynamic cohort (22 surveys, from June 1996 to June 2001) on about 1300 children (<12 years) distributed between 173 households localized by GPS. We used the computed parasitological data to analyzed levels of Plasmodium falciparum, P. malariae and P. ovale infection and P. falciparum gametocyte carriage by means of time series and Kulldorff's scan statistic for space-time cluster detection. RESULTS: The time series analysis determined that malaria parasitemia (primarily P. falciparum) was persistently present throughout the population with the expected seasonal variability pattern and a downward temporal trend. We identified six high-risk clusters of P. falciparum infection, some of which persisted despite an overall tendency towards a decrease in risk. The first high-risk cluster of P. falciparum infection (rate ratio = 14.161) was detected from September 1996 to October 1996, in the north of the village. CONCLUSION: This study showed that, although infection proportions tended to decrease, high risk zones persisted in the village particularly near temporal backwaters. Analysis of this heterogeneity at the household scale by GIS methods lead to target preventive actions more accurately on the high-risk zones identified. This mapping of malaria risk makes it possible to orient control programs, treating the high-risk zones identified as a matter of priority, and to improve the planning of intervention trials or research studies on malaria. PMID- 17118177 TI - IGF-II transgenic mice display increased aberrant colon crypt multiplicity and tumor volume after 1,2-dimethylhydrazine treatment. AB - In colorectal cancer insulin-like growth factor II (IGF-II) is frequently overexpressed. To evaluate, whether IGF-II affects different stages of tumorigenesis, we induced neoplastic alterations in the colon of wild-type and IGF-II transgenic mice using 1,2-dimethylhydrazine (DMH). Aberrant crypt foci (ACF) served as markers of early lesions in the colonic mucosa, whereas adenomas and carcinomas characterized the endpoints of tumor development. DMH-treatment led initially to significantly more ACF in IGF-II transgenic than in wild-type mice. This increase in ACF was especially prominent for those consisting of > or =three aberrant crypts (AC). Nevertheless, adenomas and adenocarcinomas of the colon, present after 34 weeks in both genetic groups, were not found at different frequency. Tumor volumes, however, were significantly higher in IGF-II transgenic mice and correlated with serum IGF-II levels. Immunohistochemical staining for markers of proliferation and apoptosis revealed increased cell proliferation rates in tumors of IGF-II transgenic mice without significant affection of apoptosis. Increased proliferation was accompanied by elevated localization of beta-catenin in the cytosol and cell nuclei and reduced appearance at the inner plasma membrane. In conclusion, we provide evidence that IGF-II, via activation of the beta-catenin signaling cascade, promotes growth of ACF and tumors without affecting tumor numbers. PMID- 17118178 TI - A rare case of sarcoidosis involving the middle turbinates: an incidental diagnosis. AB - BACKGROUND: Sarcoidosis is a chronic, systemic granulomatous disease of unknown etiology that features noncaseating granulomas in many body regions. Sinonasal involvement is rare but is also suspected to be underreported. CASE PRESENTATION: We present the case of a 39-year-old woman who was incidentally diagnosed with isolated sarcoidosis involving the middle turbinates. Histopathologic examination of resected concha bullosa material and an extensive panel of diagnostic tests revealed a diagnosis of isolated sarcoidosis. Since no systemic manifestations were detected, topical corticosteroid (nasal spray) was administered in the postoperative period. Throughout the 12 months after surgery, the patient remained free of symptoms and all nasal endoscopy examinations were normal. CONCLUSION: Although isolated nasal involvement of sarcoidosis is rare, otorhinolaryngologists should consider this condition in a differential diagnosis for sinonasal complaints. PMID- 17118179 TI - Molecular surveillance of mutations in the cytochrome b gene of Plasmodium falciparum in Gabon and Ethiopia. AB - BACKGROUND: Atovaquone is part of the antimalarial drug combination atovaquone proguanil (Malarone) and inhibits the cytochrome bc1 complex of the electron transport chain in Plasmodium spp. Molecular modelling showed that amino acid mutations are clustered around a putative atovaquone-binding site resulting in a reduced binding affinity of atovaquone for plasmodial cytochrome b, thus resulting in drug resistance. METHODS: The prevalence of cytochrome b point mutations possibly conferring atovaquone resistance in Plasmodium falciparum isolates in atovaquone treatment-naive patient cohorts from Lambarene, Gabon and from South Western Ethiopia was assessed. RESULTS: Four/40 (10%) mutant types (four different single polymorphisms, one leading to an amino acid change from M to I in a single case) in Gabonese isolates, but all 141/141 isolates were wild type in Ethiopia were found. CONCLUSION: In the absence of drug pressure, spontaneous and possibly resistance-conferring mutations are rare. PMID- 17118180 TI - Do self- reported intentions predict clinicians' behaviour: a systematic review. AB - BACKGROUND: Implementation research is the scientific study of methods to promote the systematic uptake of clinical research findings into routine clinical practice. Several interventions have been shown to be effective in changing health care professionals' behaviour, but heterogeneity within interventions, targeted behaviours, and study settings make generalisation difficult. Therefore, it is necessary to identify the 'active ingredients' in professional behaviour change strategies. Theories of human behaviour that feature an individual's "intention" to do something as the most immediate predictor of their behaviour have proved to be useful in non-clinical populations. As clinical practice is a form of human behaviour such theories may offer a basis for developing a scientific rationale for the choice of intervention to use in the implementation of new practice. The aim of this review was to explore the relationship between intention and behaviour in clinicians and how this compares to the intention behaviour relationship in studies of non-clinicians. METHODS: We searched: PsycINFO, MEDLINE, EMBASE, CINAHL, Cochrane Central Register of Controlled Trials, Science/Social science citation index, Current contents (social & behavioural med/clinical med), ISI conference proceedings, and Index to Theses. The reference lists of all included papers were checked manually. Studies were eligible for inclusion if they had: examined a clinical behaviour within a clinical context, included measures of both intention and behaviour, measured behaviour after intention, and explored this relationship quantitatively. All titles and abstracts retrieved by electronic searching were screened independently by two reviewers, with disagreements resolved by discussion. DISCUSSION: Ten studies were found that examined the relationship between intention and clinical behaviours in 1623 health professionals. The proportion of variance in behaviour explained by intention was of a similar magnitude to that found in the literature relating to non-health professionals. This was more consistently the case for studies in which intention-behaviour correspondence was good and behaviour was self-reported. Though firm conclusions are limited by a smaller literature, our findings are consistent with that of the non-health professional literature. This review, viewed in the context of the larger populations of studies, provides encouragement for the contention that there is a predictable relationship between the intentions of a health professional and their subsequent behaviour. However, there remain significant methodological challenges. PMID- 17118181 TI - Improving the use of research evidence in guideline development: 1. Guidelines for guidelines. AB - BACKGROUND: The World Health Organization (WHO), like many other organisations around the world, has recognised the need to use more rigorous processes to ensure that health care recommendations are informed by the best available research evidence. This is the first of a series of 16 reviews that have been prepared as background for advice from the WHO Advisory Committee on Health Research to WHO on how to achieve this. OBJECTIVES: We reviewed the literature on guidelines for the development of guidelines. METHODS: We searched PubMed and three databases of methodological studies for existing systematic reviews and relevant methodological research. We did not conduct systematic reviews ourselves. Our conclusions are based on the available evidence, consideration of what WHO and other organisations are doing and logical arguments. KEY QUESTIONS AND ANSWERS: We found no experimental research that compared different formats of guidelines for guidelines or studies that compared different components of guidelines for guidelines. However, there are many examples, surveys and other observational studies that compared the impact of different guideline development documents on guideline quality. WHAT HAVE OTHER ORGANIZATIONS DONE TO DEVELOP GUIDELINES FOR GUIDELINES FROM WHICH WHO CAN LEARN?: Establish a credible, independent committee that evaluates existing methods for developing guidelines or that updates existing ones. Obtain feedback and approval from various stakeholders during the development process of guidelines for guidelines. Develop a detailed source document (manual) that guideline developers can use as reference material. WHAT SHOULD BE THE KEY COMPONENTS OF WHO GUIDELINES FOR GUIDELINES?: Guidelines for guidelines should include information and instructions about the following components: 1) Priority setting; 2) Group composition and consultations; 3) Declaration and avoidance of conflicts of interest; 4) Group processes; 5) Identification of important outcomes; 6) Explicit definition of the questions and eligibility criteria ; 7) Type of study designs for different questions; 8) Identification of evidence; 9) Synthesis and presentation of evidence; 10) Specification and integration of values; 11) Making judgments about desirable and undesirable effects; 12) Taking account of equity; 13) Grading evidence and recommendations; 14) Taking account of costs; 15) Adaptation, applicability, transferability of guidelines; 16) Structure of reports; 17) Methods of peer review; 18) Planned methods of dissemination & implementation; 19) Evaluation of the guidelines. WHAT HAVE OTHER ORGANIZATIONS DONE TO IMPLEMENT GUIDELINES FOR GUIDELINES FROM WHICH WHO CAN LEARN?: Obtain buy in from regions and country level representatives for guidelines for guidelines before dissemination of a revised version. Disseminate the guidelines for guidelines widely and make them available (e.g. on the Internet). Develop examples of guidelines that guideline developers can use as models when applying the guidelines for guidelines. Ensure training sessions for those responsible for developing guidelines. Continue to monitor the methodological literature on guideline development. PMID- 17118182 TI - Variable coordination of cotranscribed genes in Escherichia coli following antisense repression. AB - BACKGROUND: A majority of bacterial genes belong to tight clusters and operons, which complicates gene functional studies using conventional knock-out methods. Antisense agents can down-regulate the expression of genes without disrupting the genome because they bind mRNA and block its expression. However, it is unclear how antisense inhibition affects expression from genes that are cotranscribed with the target. RESULTS: To examine the effects of antisense inhibition on cotranscribed genes, we constructed a plasmid expressing the two reporter genes gfp and DsRed as one transcriptional unit. Incubation with antisense peptide nucleic acid (PNA) targeted to the mRNA start codon region of either the upstream gfp or the downstream DsRed gene resulted in a complete expression discoordination from this artificial construct. The same approach was applied to the three cotranscribed genes in the endogenously expressed lac-operon (lacZ, Y and A) and partial downstream expression coordination was seen when the lacZ start codon was targeted with antisense PNA. Targeting the lacY mRNA start codon region showed no effect on the upstream lacZ gene expression whereas expression from the downstream lacA gene was affected as strongly as the lacY gene. Determination of lacZ and lacY mRNA levels revealed a pattern of reduction that was similar to the Lac-proteins, indicating a relation between translation inhibition and mRNA degradation as a response to antisense PNA treatment. CONCLUSION: The results show that antisense mediated repression of genes within operons affect cotranscribed genes to a variable degree. Target transcript stability appears to be closely related to inhibition of translation and presumably depends on translating ribosomes protecting the mRNA from intrinsic decay mechanisms. Therefore, for genes within operons and clusters it is likely that the nature of the target transcript will determine the inhibitory effects on cotranscribed genes. Consequently, no simple and specific methods for expression control of a single gene within polycistronic operons are available, and a thorough understanding of mRNA regulation and stability is required to understand the results from both knock-down and knock-out methods used in bacteria. PMID- 17118183 TI - Coronary bypass grafting using crossclamp fibrillation does not result in reliable reperfusion of the myocardium when the crossclamp is intermittently released: a prospective cohort study. AB - BACKGROUND: Cross-clamp fibrillation is a well established method of performing coronary grafting, but its clinical effect on the myocardium is unknown. We sought to measure these effects clinically using the Khuri Intramyocardial pH monitor. METHODS: 50 episodes of cross-clamping were recorded in 16 patients who underwent CABG with crossclamp-fibrillation. An Intramyocardial pH probe measured the level of acidosis in the anterior and posterior myocardium in real-time. The pH at the start and end of each period of cross-clamping was recorded. RESULTS: It became very apparent that the pH of some patients recovered quickly while others entirely failed to recover. Thus the patients were split into 2 groups according to whether the pH recovered to above 6.8 after the first crossclamp release (N = 8 in each group). Initial pH was 7.133 (range 6.974-7.239). After the first period of crossclamping the pH dropped to 6.381 (range 6.034-6.684). The pH in recoverers prior to the second XC application was 6.990 (range 6.808 7.222) compared to only 6.455 (range 6.200-6.737) in patient's whose myocardium did not recover (P < 0.0005). This finding was repeated after the second XC release (mean pH 7.005 vs 6.537) and the third (mean pH 6.736 vs 6.376). However prior to separation from bypass the pH was close to the initial pH in both groups (7.062 vs 7.038). CONCLUSION: Crossclamp fibrillation does not result in reliable reperfusion of the myocardium between periods of crossclamping. PMID- 17118184 TI - Malignancy in the blind painful eye--report of two cases and literature review. AB - BACKGROUND: Few cases of malignant tumors arising in a blind painful eye have previously been described. We described two cases of a blind painful eye containing an unsuspected tumor, which were enucleated to relieve the pain. CASE PRESENTATIONS: Case 1: A 57 year-old Caucasian man presented with recurrent orbital cellulitis and endophthalmitis in the left eye (OS). The OS was blind and painful and an enucleation was performed showing a uveal melanoma by histopathological exam. Case 2: A 54 year-old Caucasian man with previous history of a rhegmatogenous retinal detachment in his left eye presented a blind painful eye. Enucleation was performed revealing a well-differentiated B-cell lymphoma of uveal tract with extra ocular extension. CONCLUSION: In the management of a blind painful eye, it is extremely important to rule out an intraocular malignancy particularly in those patients who have not been followed by an ophthalmologist. PMID- 17118185 TI - Giant solitary fibrous tumour of the liver. AB - BACKGROUND: Solitary fibrous tumour (SFT) is an uncommon mesenchymal neoplasm that most frequently affects the pleura, although it has been reported with increasing frequency in various other sites such as in the peritoneum, pericardium and in non-serosal sites such as lung parenchyma, upper respiratory tract, orbit, thyroid, parotid gland, or thymus. Liver parenchyma is rarely affected. Clinically, SFTs cause symptoms after having reached a certain size or when vital structures are involved. In recent years, SFTs are more often identified and distinguished from other tumours with a similar appearance due to the availability of characteristic immunohistochemical markers. CASE PRESENTATION: In this manuscript we report the case of a large tumour of the liver, which was histologically diagnosed as a SFT, and showed involvement of a single hepatic segment. Because of the patient's presentation and clinical course, it may represent a radiation-induced lesion. CONCLUSION: When a SFT has been diagnosed, surgery is the treatment of choice. The small number of patients with a SFT of the liver and its unknown natural behaviour creates the need to a careful registration and follow-up of all identified cases. PMID- 17118186 TI - Comparison of uncalibrated arterial waveform analysis in cardiac surgery patients with thermodilution cardiac output measurements. AB - INTRODUCTION: Cardiac output (CO) monitoring is indicated only in selected patients. In cardiac surgical patients, perioperative haemodynamic management is often guided by CO measurement by pulmonary artery catheterisation (COPAC). Alternative strategies of CO determination have become increasingly accepted in clinical practice because the benefit of guiding therapy by data derived from the PAC remains to be proven and less invasive alternatives are available. Recently, a device offering uncalibrated CO measurement by arterial waveform analysis (COWave) was introduced. As far as this approach is concerned, however, the validity of the CO measurements obtained is utterly unclear. Therefore, the aim of this study was to compare the bias and the limits of agreement (LOAs) (two standard deviations) of COWave at four specified time points prior, during, and after coronary artery bypass graft (CABG) surgery with a simultaneous measurement of the gold standard COPAC and aortic transpulmonary thermodilution CO (COTranspulm). METHODS: Data from 30 patients were analysed during this prospective study. COPAC, COTranspulm, and COWave were determined in all patients at four different time points prior, during, and after CABG surgery. The COPAC and the COTranspulm were measured by triple injection of 10 ml of iced isotone sodium chloride solution into the central venous line of the PAC. Measurements of COWave were simultaneously taken at these time points. RESULTS: The overall correlation showed a Spearman correlation coefficient between COPAC and COWave of 0.53 (p < 0.01) and 0.84 (p < 0.01) for COPAC and COTranspulm. Bland-Altman analysis showed a mean bias and LOAs of 0.6 litres per minute and -2.2 to +3.4 litres per minute for COPAC versus COWave and -0.1 litres per minute and -1.8 to +1.6 litres per minute for COPAC versus COTranspulm. CONCLUSION: Arterial waveform analysis with an uncalibrated algorithm COWave underestimated COPAC to a clinically relevant extent. The wide range of LOAs requires further evaluation. Better results might be achieved with an improved new algorithm. In contrast to this, we observed a better correlation of thermodilution COTranspulm and thermodilution COPAC measurements prior, during, and after CABG surgery. PMID- 17118187 TI - The competent sentinel node: an association with an axillary presentation and an occult or a small primary invasive breast carcinoma. AB - The concept of the sentinel node describes a primary or sentinel lymph node (SLN), which exists and through which tumour cells from a primary tumour in a particular location must first travel to spread to a particular regional lymph node group. In this series we present three patients presenting with a pathological axillary node associated with either an occult or very small primary breast cancer. In each case the primary tumour was found to have metastasised to the palpable node, however despite the significant enlargement of this node, no other axillary nodes were found to be affected on axillary node clearance. This has led us to postulate that the SLN in some cases contains unique characteristics that enable it to prevent further spread of the tumour up the lymphatic chain. Hence the term the competent sentinel node. PMID- 17118188 TI - Acute tubulo-interstitial nephritis leading to acute renal failure following multiple hornet stings. AB - BACKGROUND: Hornet stings are generally associated with local and occasionally anaphylactic reactions. Rarely systemic complications like acute renal failure can occur following multiple stings. Renal failure is usually due to development of acute tubular necrosis as a result of intravascular haemolysis, rhabdomyolysis or shock. Rarely it can be following development of acute tubulo-interstitial nephritis. CASE PRESENTATION: We describe a young male, who was stung on face, head, shoulders and upper limbs by multiple hornets (Vespa orientalis). He developed acute renal failure as a result of acute tubulo-interstitial nephritis and responded to steroids. CONCLUSION: Rare causes of acute renal failure like tubulo-interstitial nephritis should be considered in a patient with persistent oliguria and azotemia following multiple hornet stings. Renal biopsy should be undertaken early, as institution of steroid therapy may help in recovery of renal function. PMID- 17118189 TI - SMOTIF: efficient structured pattern and profile motif search. AB - BACKGROUND: A structured motif allows variable length gaps between several components, where each component is a simple motif, which allows either no gaps or only fixed length gaps. The motif can either be represented as a pattern or a profile (also called positional weight matrix). We propose an efficient algorithm, called SMOTIF, to solve the structured motif search problem, i.e., given one or more sequences and a structured motif, SMOTIF searches the sequences for all occurrences of the motif. Potential applications include searching for long terminal repeat (LTR) retrotransposons and composite regulatory binding sites in DNA sequences. RESULTS: SMOTIF can search for both pattern and profile motifs, and it is efficient in terms of both time and space; it outperforms SMARTFINDER, a state-of-the-art algorithm for structured motif search. Experimental results show that SMOTIF is about 7 times faster and consumes 100 times less memory than SMARTFINDER. It can effectively search for LTR retrotransposons and is well suited to searching for motifs with long range gaps. It is also successful in finding potential composite transcription factor binding sites. CONCLUSION: SMOTIF is a useful and efficient tool in searching for structured pattern and profile motifs. The algorithm is available as open-source at: http://www.cs.rpi.edu/~zaki/software/sMotif/. PMID- 17118190 TI - Connectivity independent protein-structure alignment: a hierarchical approach. AB - BACKGROUND: Protein-structure alignment is a fundamental tool to study protein function, evolution and model building. In the last decade several methods for structure alignment were introduced, but most of them ignore that structurally similar proteins can share the same spatial arrangement of secondary structure elements (SSE) but differ in the underlying polypeptide chain connectivity (non sequential SSE connectivity). RESULTS: We perform protein-structure alignment using a two-level hierarchical approach implemented in the program GANGSTA. On the first level, pair contacts and relative orientations between SSEs (i.e. alpha helices and beta-strands) are maximized with a genetic algorithm (GA). On the second level residue pair contacts from the best SSE alignments are optimized. We have tested the method on visually optimized structure alignments of protein pairs (pairwise mode) and for database scans. For a given protein structure, our method is able to detect significant structural similarity of functionally important folds with non-sequential SSE connectivity. The performance for structure alignments with strictly sequential SSE connectivity is comparable to that of other structure alignment methods. CONCLUSION: As demonstrated for several applications, GANGSTA finds meaningful protein-structure alignments independent of the SSE connectivity. GANGSTA is able to detect structural similarity of protein folds that are assigned to different superfamilies but nevertheless possess similar structures and perform related functions, even if these proteins differ in SSE connectivity. PMID- 17118191 TI - Effect of a mixture of micronutrients, but not of bovine colostrum concentrate, on immune function parameters in healthy volunteers: a randomized placebo controlled study. AB - BACKGROUND: Supplementation of nutritional deficiencies helps to improve immune function and resistance to infections in malnourished subjects. However, the suggested benefits of dietary supplementation for immune function in healthy well nourished subjects is less clear. Among the food constituents frequently associated with beneficial effects on immune function are micronutrients such as vitamin C, vitamin E, beta-carotene and zinc, and colostrum. This study was designed to investigate the effects these ingredients on immune function markers in healthy volunteers. METHODS: In a double-blind, randomized, parallel, 2*2, placebo-controlled intervention study one hundred thirty-eight healthy volunteers aged 40-80 y (average 57 +/- 10 y) received one of the following treatments: (1) bovine colostrum concentrate 1.2 g/d (equivalent to approximately 500 mg/d immunoglobulins), (2) micronutrient mix of 288 mg vitamin E, 375 mg vitamin C, 12 mg beta-carotene and 15 mg zinc/day, (3) combination of colostrum and micronutrient mix, or (4) placebo. Several immune function parameters were assessed after 6 and 10 weeks. Data were analyzed by analysis of variance. Groups were combined to test micronutrient treatment versus no micronutrient treatment, and colostrum treatment versus no colostrum treatment. RESULTS: Overall, consumption of the micronutrient mix significantly enhanced delayed-type hypersensitivity (DTH) responses (p < 0.05). Adjusted covariance analysis showed a positive association between DTH and age. Separate analysis of younger and older age groups indicated that it was the older population that benefited from micronutrient consumption. The other immune function parameters including responses to systemic tetanus and oral typhoid vaccination, phagocytosis, oxidative burst, lymphocyte proliferation and lymphocyte subset distribution were neither affected by the consumption of micronutrients nor by the consumption of bovine colostrum concentrate. CONCLUSION: Consumption of bovine colostrum had no effect on any of the immune parameters assessed. The micronutrient mix enhanced cellular immunity as measured by DTH, with an increased effect by incremental age, but did not affect any of the other immune parameters measured. Although correlations between decreased DTH and enhanced risk of certain infection have been reported, it remains unclear whether and enhanced DTH response actually improves immune defense. The present data suggests that improvement of immune parameters in a population with a generally good immune and nutritional status is limited and that improvement of immune function in this population may be difficult. PMID- 17118192 TI - A self-inactivating retrovector incorporating the IL-2 promoter for activation induced transgene expression in genetically engineered T-cells. AB - BACKGROUND: T-cell activation leads to signaling pathways that ultimately result in induction of gene transcription from the interleukin-2 (IL-2) promoter. We hypothesized that the IL-2 promoter or its synthetic derivatives can lead to T cell specific, activation-induced transgene expression. Our objective was to develop a retroviral vector for stable and activation-induced transgene expression in T-lymphocytes. RESULTS: First, we compared the transcriptional potency of the full-length IL-2 promoter with that of a synthetic promoter composed of 3 repeats of the Nuclear Factor of Activated T-Cells (NFAT) element following activation of transfected Jurkat T-cells expressing the large SV40 T antigen (Jurkat TAg). Although the NFAT3 promoter resulted in a stronger induction of luciferase reporter expression post stimulation, the basal levels of the IL-2 promoter-driven reporter expression were much lower indicating that the IL-2 promoter can serve as a more stringent activation-dependent promoter in T cells. Based on this data, we generated a self-inactivating retroviral vector with the full-length human IL-2 promoter, namely SINIL-2pr that incorporated the enhanced green fluorescent protein (EGFP) fused to herpes simplex virus thymidine kinase as a reporter/suicide "bifunctional" gene. Subsequently, Vesicular Stomatitis Virus-G Protein pseudotyped retroparticles were generated for SINIL 2pr and used to transduce the Jurkat T-cell line and the ZAP-70-deficient P116 cell line. Flow cytometry analysis showed that EGFP expression was markedly enhanced post co-stimulation of the gene-modified cells with 1 muM ionomycin and 10 ng/ml phorbol 12-myristate 13-acetate (PMA). This activation-induced expression was abrogated when the cells were pretreated with 300 nM cyclosporin A. CONCLUSION: These results demonstrate that the SINIL-2pr retrovector leads to activation-inducible transgene expression in Jurkat T-cell lines. We propose that this design can be potentially exploited in several cellular immunotherapy applications. PMID- 17118193 TI - Responsibility, opportunity, and vision for higher education in urban and regional carbon management. AB - This is a summary of the conversation among scholars attending the special session on "Responsibility, Opportunity, and Vision for Higher Education in Urban and Regional Carbon Management" at the First International Conference on Carbon Management at Urban and Regional Levels: Connecting Development Decisions to Global Issues in Mexico City Sept. 4-8, 2006. It includes The Declaration for Carbon Management Education, agreed upon by the participants. Obstacles to such a vision were discussed along with exemplar models of transdisciplinary curricula and suggestions for scholarship. PMID- 17118194 TI - Emerging aspects of oesophageal and gastro-oesophageal junction cancer histopathology - an update for the surgical oncologist. AB - Adenocarcinoma of the oesophagus and gastro-oesophageal junction are rapidly increasing in incidence and have a well described sequence of carcinogenesis: the Barrett's metaplasia-dysplasia-adenocarcinoma sequence. During recent years there have been changes in the knowledge surrounding disease progression, cancer management and histopathology specimen reporting. Tumours around the gastro oesophageal junction (GOJ) pose several specific challenges. Numerous difficulties arise when the existing TNM staging systems for gastric and oesophageal cancers are applied to GOJ tumours. The issues facing the current TNM staging and GOJ tumour classification systems are reviewed in this article. Recent evidence regarding the importance of several histopathologically derived prognostic factors, such as circumferential resection margin status and lymph node metastases, have implications for specimen reporting. With the rising use of multimodal treatments for oesophageal cancer it is important that the response of the tumour to this therapy is carefully documented pathologically. In addition, several controversial and novel areas such as endoscopic mucosal resection, lymph node micrometastases and the sentinel node concept are being studied. We aim to review these aspects, with special relevance to oesophageal and gastro oesophageal cancer specimen reporting, to update the surgical oncologist with an interest in upper gastrointestinal cancer. PMID- 17118195 TI - Lethality of suicidal organophosphorus poisoning in an Indian population: exploring preventability. AB - BACKGROUND: Suicide by organophosphorus poisoning is common in India. Study of factors associated with lethality may suggest methods for prevention. METHODS: Severity of symptoms, biochemical manifestation of poisoning, degree of lethality and the outcome were studied with an aim to explore the modifiable factors associated with lethality and to discuss preventability. Clinical variables were collected; symptoms were rated by the physicians using global impression of severity; and the lethality was assessed by scale for assessment of lethality of suicide attempt (SALSA), in 100 consecutive patients with suicidal organophosphorus poisoning attending a medical college hospital in South India. RESULTS: Fatal outcome (n = 26) was significantly associated with higher mean age, lower mean pseudocholinesterase level, longer duration between organophosphorus compound ingestion and specific intervention. All those who died had respiratory failure. Physicians' assessment of symptom severity and lethality as assessed by the SALSA could differentiate those succumbed and survived in a significant proportion. CONCLUSION: Majority of cases of organophosphorus poisoning were associated with severe symptoms and higher lethality. Intervention facilities decreasing the period between the ingestion of poison and initiation of treatment might prevent many deaths. Measures like restricting availability and banning more toxic organophosphorus compounds may help. PMID- 17118196 TI - Direct and indirect effects of retinoic acid on human Th2 cytokine and chemokine expression by human T lymphocytes. AB - BACKGROUND: Vitamin A (VA) deficiency induces a type 1 cytokine response and exogenously provided retinoids can induce a type 2 cytokine response both in vitro and in vivo. The precise mechanism(s) involved in this phenotypic switch are inconsistent and have been poorly characterized in humans. In an effort to determine if retinoids are capable of inducing Th2 cytokine responses in human T cell cultures, we stimulated human PBMCs with immobilized anti-CD3 mAb in the presence or absence of all-trans retinoic acid (ATRA) or 9-cis-RA. RESULTS: Stimulation of human PBMCs and purified T cells with ATRA and 9-cis-RA increased mRNA and protein levels of IL-4, IL-5, and IL-13 and decreased levels of IFN gamma, IL-2, IL-12p70 and TNF-alpha upon activation with anti-CD3 and/or anti CD28 mAbs. These effects were dose-dependent and evident as early as 12 hr post stimulation. Real time RT-PCR analysis revealed a dampened expression of the Th1 associated gene, T-bet, and a time-dependent increase in the mRNA for the Th2 associated genes, GATA-3, c-MAF and STAT6, upon treatment with ATRA. Besides Th1 and Th2 cytokines, a number of additional proinflammatory and regulatory cytokines including several chemokines were also differentially regulated by ATRA treatment. CONCLUSION: These data provide strong evidence for multiple inductive roles for retinoids in the development of human type-2 cytokine responses. PMID- 17118197 TI - Evaluating complementary and alternative medicine interventions: in search of appropriate patient-centered outcome measures. AB - BACKGROUND: Central to the development of a sound evidence base for Complementary and Alternative Medicine (CAM) interventions is the need for valid, reliable and relevant outcome measures to assess whether the interventions work. We assessed the specific needs for a database that would cover a wide range of outcomes measures for CAM research and considered a framework for such a database. METHODS: The study was a survey of CAM researchers, practitioners and students. An online questionnaire was emailed to the members of the Canadian Interdisciplinary Network for CAM Research (IN-CAM) and the CAM Education and Research Network of Alberta (CAMera). The majority of survey questions were open ended and asked about outcome measures currently used, outcome measures' assessment criteria, sources of information, perceived barriers to finding outcome measures and outcome domains of importance. Descriptive quantitative analysis and qualitative content analysis were used. RESULTS: One hundred and sixty-four completed surveys were received. Of these, 62 respondents reported using outcome measures in their CAM research and identified 92 different specific outcomes. The most important barriers were the fact that, for many health concepts, outcome measures do not yet exist, as well as issues related to accessibility of instruments. Important outcome domains identified included physical, psychological, social, spiritual, quality of life and holistic measures. Participants also mentioned the importance of individualized measures that assess unique patient-centered outcomes for each research participant, and measures to assess the context of healing and the process of healing. CONCLUSION: We have developed a preliminary framework that includes all components of health related outcomes. The framework provides a foundation for a larger, comprehensive collection of CAM outcomes. It fits very well in a whole systems perspective, which requires an expanded set of outcome measures, such as individualized and holistic measures, with attention to issues of process and context. PMID- 17118198 TI - Investigation of de novo cholesterol synthetic capacity in the gonads of goldfish (Carassius auratus) exposed to the phytosterol beta-sitosterol. AB - Total and intra-mitochondrial gonadal cholesterol concentrations are decreased in fish exposed to the phytoestrogen beta-sitosterol (beta-sit). The present study examined the potential for beta-sit to disrupt de novo cholesterol synthesis in the gonads of goldfish exposed to 200 microgram/g beta-sit and 10 microgram/g 17beta-estradiol (E2; estrogenic control) by intra-peritoneal Silastic implants for 21 days. The de novo cholesterol synthetic capacity was estimated by incubating gonadal tissue with 14C-acetate for a period of 18 hours, followed by chloroform/methanol lipid extraction and thin layer chromatography (TLC) lipid separation. Lipid classes were confirmed using infrared spectroscopy. Plasma testosterone (T) and total cholesterol concentration were measured and gonadosomatic index (GSI) was calculated. Plasma T was significantly reduced in male beta-sit-treated fish compared to control and E2-treated fish (p < 0.001). 14C-Acetate incorporation into cholesterol and cholesterol esters was not significantly different among treatment groups for male and female fish, however, 14C-enrichment was higher than expected in both triglycerides (TG) and free fatty acids (FFA). FFA incorporation was significantly higher in male control fish than either beta-sit or E2 treatments (p = 0.005). Plasma cholesterol concentration was significantly increased in the male beta-sit treatment group compared to controls (p = 0.027). These results indicate gonadal de novo cholesterol biosynthetic capacity is not disrupted by beta-sit or E2 treatment in early recrudescing male or female goldfish, while plasma cholesterol and steroid concentrations are sensitive to beta-sit exposure. PMID- 17118199 TI - Molecular evidence of Ureaplasma urealyticum and Ureaplasma parvum colonization in preterm infants during respiratory distress syndrome. AB - BACKGROUND: Ureaplasma urealyticum and U. parvum have been associated with respiratory diseases in premature newborns, but their role in the pathogenesis of the respiratory distress syndrome (RDS) is unclear. The aim of this study was to detect, using molecular techniques, the role of Mycoplasma spp. and Ureaplasma spp. in respiratory secretion and blood specimens of preterm newborns with or without RDS and to evaluate the prevalence of perinatal U. urealyticum or U. parvum infection. The influence of chemotherapy on the clinical course was also evaluated. METHODS: Tracheal aspirate or nasopharingeal fluid samples from 50 preterm babies with (24) or without RDS (26) were analysed for detection of U. urealyticum and U. parvum by culture identification assay and PCR. Sequencing analysis of amplicons allowed us to verify the specificity of methods. Clarithromycin (10 mg kg-1 twice a day) was administered in ureaplasma-positive patients who presented clinical signs of RDS. RESULTS: 15/24 neonates with RDS (p < 0.001) and 4/26 without RDS were found PCR-positive for U. urealyticum or U. parvum. Culture identification assay was positive in 5/50 newborns, three of which with RDS. Sequencing analyses confirmed the specificity of these methods. Association of patent ductus arteriosus with ureaplasma colonization was more statistically significant (p = 0.0004) in patients with RDS than in those without RDS. CONCLUSION: Colonization of the lower respiratory tract by Ureaplasma spp. and particularly by U. parvum in preterm newborns was related to RDS. The routine use of molecular methods could be useful to screen candidate babies for etiologic therapy. PMID- 17118200 TI - Incomplete oedipism and chronic suicidality in psychotic depression with paranoid delusions related to eyes. AB - Self-enucleation or oedipism is a term used to describe self-inflicted enucleation. It is a rare form of self-mutilation, found mainly in acutely psychotic patients. We propose the term incomplete oedipism to describe patients who deliberately and severely mutilate their eyes without proper enucleation. We report the case of a 32-year-old male patient with a five-year history of psychotic depression accompanied by paranoid delusions centered around his belief that his neighbors criticized him and stared at him. A central feature of his clinical picture was an eye injury that the patient had caused by pouring molten lead into his right eye during a period of deep hopelessness and suicidality when the patient could not resolve his anhedonia and social isolation. Pharmacotherapy and psychotherapy dramatically improved his disorder. PMID- 17118201 TI - Pharmacological inhibition of leukotrienes in an animal model of bleomycin induced acute lung injury. AB - Leukotrienes are increased locally in idiopathic pulmonary fibrosis. Furthermore, a role for these arachidonic acid metabolites has been thoroughly characterized in the animal bleomycin model of lung fibrosis by using different gene knock-out settings. We investigated the efficacy of pharmacological inhibition of leukotrienes activity in the development of bleomycin-induced lung injury by comparing the responses in wild-type mice with mice treated with zileuton, a 5 lipoxygenase inhibitor and MK-571, a cys-leukotrienes receptor antagonist. Mice were subjected to intra-tracheal administration of bleomycin or saline and were assigned to receive either MK-571 at 1 mg/Kg or zileuton at 50 mg/Kg daily. One week after bleomycin administration, BAL cell counts, lung histology with van Gieson for collagen staining and immunohistochemical analysis for myeloperoxidase, IL-1 and TNF-alpha were performed. Following bleomycin administration both MK-571 and zileuton treated mice exhibited a reduced degree of lung damage and inflammation when compared to WT mice as shown by the reduction of:(i) loss of body weight, (ii) mortality rate, (iii) lung infiltration by neutrophils (myeloperoxidase activity, BAL total and differential cell counts), (iv) lung edema, (v) histological evidence of lung injury and collagen deposition, (vi) lung myeloperoxidase, IL-1 and TNF-alpha staining. This is the first study showing that the pharmacological inhibition of leukotrienes activity attenuates bleomycin-induced lung injury in mice. Given our results as well as those coming from genetic studies, it might be considered meaningful to trial this drug class in the treatment of pulmonary fibrosis, a disease that still represents a major challenge to medical treatment. PMID- 17118202 TI - The effect of the Thanksgiving holiday on weight gain. AB - BACKGROUND: More people than ever are considered obese and the resulting health problems are evident. These facts highlight the need for identification of critical time periods for weight gain. Therefore the purpose was to assess potential changes that occur in body weight during the Thanksgiving holiday break in college students. METHODS: 94 college students (23.0 +/- 4.6 yrs, 72.1 +/- 14.0 kg, 172.6 +/- 9.3 cm, 24.0 +/- 3.9 kg/m2) reported to the human body composition laboratory at the University of Oklahoma following a 6-hour fast with testing occurring prior to, and immediately following the Thanksgiving holiday break (13 +/- 3 days). Body weight (BW) was assessed using a balance beam scale while participants were dressed in minimal clothing. Paired t-tests were used to assess changes in BW pre and post Thanksgiving holiday with additional analysis by gender, body mass index (BMI), and class standing (i.e. undergraduate vs. graduate). RESULTS: Overall, a significant (P < 0.05) increase in BW was found between pre (72.1 kg) and post (72.6 kg) Thanksgiving holiday. When stratified by gender and class standing a significant (P < 0.05) increase in body weight was observed between the pre and post Thanksgiving holiday in males (0.6 kg), females (0.4 kg) and graduate students (0.8 kg). When participants were classified by BMI as normal or as overweight/obese, a significant 1.0 kg BW gain was found (P < 0.05) in the overweight/obese (>/=25 kg/m2) group compared to a non significant 0.2 kg gain in the normal group (<25 kg/m2). CONCLUSION: These data indicate that participants in our study gained a significant amount of BW (0.5 kg) during the Thanksgiving holiday. While an increase in BW of half a kilogram may not be cause for alarm, the increase could have potential long-term health consequences if participants retained this weight gain throughout the college year. Additionally, because the overweight/obese participants gained the greatest amount of BW, this group may be at increased risk for weight gain and further obesity development during the holiday season. PMID- 17118203 TI - The separate and combined effects of MHC genotype, parasite clone, and host gender on the course of malaria in mice. AB - BACKGROUND: The link between host MHC (major histocompatibility complex) genotype and malaria is largely based on correlative data with little or no experimental control of potential confounding factors. We used an experimental mouse model to test for main effects of MHC-haplotypes, MHC heterozygosity, and MHC x parasite clone interactions. We experimentally infected MHC-congenic mice (F2 segregants, homo- and heterozygotes, males and females) with one of two clones of Plasmodium chabaudi and recorded disease progression. RESULTS: We found that MHC haplotype and parasite clone each have a significant influence on the course of the disease, but there was no significant host genotype by parasite genotype interaction. We found no evidence for overdominance nor any other sort of heterozygote advantage or disadvantage. CONCLUSION: When tested under experimental conditions, variation in the MHC can significantly influence the course of malaria. However, MHC heterozygote advantage through overdominance or dominance of resistance cannot be assumed in the case of single-strain infections. Future studies might focus on the interaction between MHC heterozygosity and multiple-clone infections. PMID- 17118204 TI - The crossroads of GIS and health information: a workshop on developing a research agenda to improve cancer control. AB - Cancer control researchers seek to reduce the burden of cancer by studying interventions, their impact in defined populations, and the means by which they can be better used. The first step in cancer control is identifying where the cancer burden is elevated, which suggests locations where interventions are needed. Geographic information systems (GIS) and other spatial analytic methods provide such a solution and thus can play a major role in cancer control. This report presents findings from a workshop held June 16-17, 2005, to bring together experts and stakeholders to address current issues in GIScience and cancer control. A broad range of areas of expertise and interest was represented, including epidemiology, geography, statistics, environmental health, social science, cancer control, cancer registry operations, and cancer advocacy. The goals of this workshop were to build consensus on important policy and research questions, identify roadblocks to future progress in this field, and provide recommendations to overcome these roadblocks. PMID- 17118205 TI - A prospective study comparing quantitative Cytomegalovirus (CMV) polymerase chain reaction in plasma and pp65 antigenemia assay in monitoring patients after allogeneic stem cell transplantation. AB - BACKGROUND: Low levels of Cytomegalovirus (CMV) viral load are frequently detected following allogeneic stem cell transplantation (SCT) and CMV disease may still develop in some allogeneic SCT patients who have negative pp65-antigenemia (pp65-Ag) or undetectable DNA. Pp65Ag is a sensitive method to diagnose CMV infection. Quantitative CMV-DNA PCR assay in plasma has been proposed to monitor CMV infection in SCT patients. We evaluated the clinical utility of pp65Ag and PCR assay in plasma of SCT recipients. METHODS: In a prospective longitudinal study, 38 consecutive patients at risk of CMV infection (donor and/or recipient CMV seropositive) were weekly monitored for CMV infection by both quantitative CMV-PCR in plasma (COBAS AMPLICOR CMV MONITOR) and pp65 Ag, during the first 100 days after SCT. RESULTS: A total of 534 blood samples were simultaneously analysed for pp65Ag and PCR. Overall, 28/38 patients (74%) had active CMV infection within 100 days from SCT. In 16 patients, CMV was first detected by pp65 Ag alone; in 5 patients by both methods and in 6 by PCR assay alone; one patient had CMV biopsy-proven intestinal disease without pp65Ag and PCR assays positivity before CMV disease. Overall, three patients developed intestinal CMV disease (7.9%): one had negative both pp65Ag and PCR assays before CMV disease, one had disease and concomitant positivity of both methods, while in the remaining patient, only pp65Ag was positive before CMV disease. CONCLUSION: Plasma PCR(COBAS AMPLICOR CMV MONITOR) and pp65Ag assays were effective in detecting CMV infection, however, discordance between both methods were frequently observed. Plasma PCR and pp65Ag assays may be complementary for diagnosis and management of CMV infection. PMID- 17118206 TI - Effects of in vitro fertilization and embryo culture on TRP53 and Bax expression in B6 mouse embryos. AB - In the mouse, embryo culture results in a characteristic phenotype of retarded embryo preimplantation development and reduced numbers of cells within embryos. The expression of TRP53 is central to the regulation of the cell's capacity to proliferate and survive. In this study we found that Trp53 mRNA is expressed throughout the preimplantation stage of development. Levels of TRP53 protein expression were low during the cleavage stages and increased at the morula and blastocyst stages in B6 embryos collected from the reproductive tract. Embryos collected at the zygote stage and cultured for 96 h also showed low levels of TRP53 expression at precompaction stages. There were higher levels of TRP53 in cultured morula and the level in cultured blastocysts was clearly increased above blastocysts collected directly from the uterus. Immunolocalization of TRP53 showed that its increased expression in cultured blastocysts corresponded with a marked accumulation of TRP53 within the nuclei of embryonic cells. This pattern of expression was enhanced in embryos produced by in vitro fertilization and subjected to culture. The TRP53 was transcriptionally active since culture also induced increased expression of Bax, yet this did not occur in embryos lacking Trp53 (Trp53-/-). The rate of development of Trp53-/- zygotes to the blastocyst stage was not different to wildtype controls when embryos were cultured in groups of ten but was significantly faster when cultured individually. The results show that zygote culture resulted in the accumulation of transcription activity of TRP53 in the resulting blastocysts. This accounts for the adverse effects of culture of embryos individually, but does not appear to be the sole cause of the retarded preimplantation stage growth phenotype associated with culture in vitro. PMID- 17118207 TI - Subacute left ventricle free wall rupture after acute myocardial infarction: awareness of the clinical signs and early use of echocardiography may be life saving. AB - Left ventricular free wall rupture (LVFWR) is a fearful complication of acute myocardial infarction in which a swift diagnosis and emergency surgery can be crucial for successful treatment. Because a significant number of cases occur subacutely, clinicians should be aware of the risk factors, clinical features and diagnostic criteria of this complication. We report the case of a 69 year-old man in whom a subacute left ventricular free wall rupture (LVFWR) was diagnosed 7 days after an inferior myocardial infarction with late reperfusion therapy. An asymptomatic 3 to 5 mm saddle-shaped ST-segment elevation in anterior and lateral leads, detected on a routine ECG, led to an urgent bedside echocardiogram which showed basal inferior-wall akinesis, a small echodense pericardial effusion and a canalicular tract from endo to pericardium, along the interface between the necrotic and normal contracting myocardium, trough which power-Doppler examination suggested blood crossing the myocardial wall. A cardiac MRI further reinforced the possibility of contained LVFWR and a surgical procedure was undertaken, confirming the diagnosis and allowing the successful repair of the myocardial tear. This case illustrates that subacute LVFWR provides an opportunity for intervention. Recognition of the diversity of presentation and prompt use of echocardiography may be life-saving. PMID- 17118208 TI - Simultaneous host and parasite expression profiling identifies tissue-specific transcriptional programs associated with susceptibility or resistance to experimental cerebral malaria. AB - BACKGROUND: The development and outcome of cerebral malaria (CM) reflects a complex interplay between parasite-expressed virulence factors and host response to infection. The murine CM model, Plasmodium berghei ANKA (PbA), which simulates many of the features of human CM, provides an excellent system to study this host/parasite interface. We designed "combination" microarrays that concurrently detect genome-wide transcripts of both PbA and mouse, and examined parasite and host transcriptional programs during infection of CM-susceptible (C57BL/6) and CM resistant (BALB/c) mice. RESULTS: Analysis of expression data from brain, lung, liver, and spleen of PbA infected mice showed that both host and parasite gene expression can be examined using a single microarray, and parasite transcripts can be detected within whole organs at a time when peripheral blood parasitemia is low. Parasites display a unique transcriptional signature in each tissue, and lung appears to be a large reservoir for metabolically active parasites. In comparisons of susceptible versus resistant animals, both host and parasite display distinct, organ-specific transcriptional profiles. Differentially expressed mouse genes were related to humoral immune response, complement activation, or cell-cell interactions. PbA displayed differential expression of genes related to biosynthetic activities. CONCLUSION: These data show that host and parasite gene expression profiles can be simultaneously analysed using a single "combination" microarray, and that both the mouse and malaria parasite display distinct tissue- and strain-specific responses during infection. This technology facilitates the dissection of host-pathogen interactions in experimental cerebral malaria and could be extended to other disease models. PMID- 17118209 TI - Statistical analysis of an RNA titration series evaluates microarray precision and sensitivity on a whole-array basis. AB - BACKGROUND: Concerns are often raised about the accuracy of microarray technologies and the degree of cross-platform agreement, but there are yet no methods which can unambiguously evaluate precision and sensitivity for these technologies on a whole-array basis. RESULTS: A methodology is described for evaluating the precision and sensitivity of whole-genome gene expression technologies such as microarrays. The method consists of an easy-to-construct titration series of RNA samples and an associated statistical analysis using non linear regression. The method evaluates the precision and responsiveness of each microarray platform on a whole-array basis, i.e., using all the probes, without the need to match probes across platforms. An experiment is conducted to assess and compare four widely used microarray platforms. All four platforms are shown to have satisfactory precision but the commercial platforms are superior for resolving differential expression for genes at lower expression levels. The effective precision of the two-color platforms is improved by allowing for probe specific dye-effects in the statistical model. The methodology is used to compare three data extraction algorithms for the Affymetrix platforms, demonstrating poor performance for the commonly used proprietary algorithm relative to the other algorithms. For probes which can be matched across platforms, the cross-platform variability is decomposed into within-platform and between-platform components, showing that platform disagreement is almost entirely systematic rather than due to measurement variability. CONCLUSION: The results demonstrate good precision and sensitivity for all the platforms, but highlight the need for improved probe annotation. They quantify the extent to which cross-platform measures can be expected to be less accurate than within-platform comparisons for predicting disease progression or outcome. PMID- 17118210 TI - Preliminary results of phase I trial of oral uracil/tegafur (UFT), leucovorin plus irinotecan and radiation therapy for patients with locally recurrent rectal cancer. AB - BACKGROUND: Surgical attempts for locally recurrent rectal cancer often fail due to local re-recurrence and distant metastasis. Preoperative chemoradiation may enhance better local control and survival. The aim of this study was to assess the safety of oral uracil and tegafur (UFT) plus leucovorin (LV), and irinotecan combined with radiation and determine the maximum-tolerated dose (MTD) and dose limiting toxicity (DLT) of the triple drug regimen. PATIENTS AND METHODS: Patients with locally recurrent rectal cancer received escalating doses of irinotecan on days 1, 8, 15, and 22 (starting at 30 mg/m2, with 10 mg increments between consecutive cohorts) and fixed doses of UFT (300 mg/m2) plus LV (75 mg/day) on days 3 to 7, 10 to 14, 17 to 21, and 24 to 28. Radiation was given 5 days per week totaling 40 to 50 Gy (2Gy/day). RESULTS: Six patients were treated at the starting dose, and 2 received the full scheduled chemoradiotherapy. The other 4 patients had grade 3 diarrhea and diarrhea was the DLT. One patient had partial response and he had subsequently radical surgical resection. Median progression free survival for local recurrence was 320 days. CONCLUSION: Irinotecan plus UFT/LV with concomitant radiotherapy in patients with locally recurrent rectal cancer was not feasible due to diarrhea in this setting. Modification of the treatment is needed. PMID- 17118211 TI - A study of bovine mastitis, milking procedures and management practices on 25 Estonian dairy herds. AB - BACKGROUND: Mastitis prevalence, milking procedures and management practices were investigated in 25 big dairy herds supplying milk to an Estonian dairy company. The aim of the study was to provide information for the company to be used in their new udder health improvement program to be set up after the completion of this study. METHODS: Quarter milk samples were collected from 3,166 cows for bacterial analysis and SCC (somatic cell counting). During the farm visit the veterinarian filled in a questionnaire about milking procedures and management practices with the help of farm managers. If the milk SCC of a cow or of a quarter exceeded 200,000/ml, the cow was defined as having mastitis. RESULTS: The percentage of cows having inflammation in one or more quarters measured by SCC (200,000/ml) was 52.7%. Corynebacterium bovis, Staphylococcus aureus and coagulase negative staphylococci were the most common bacterial isolates. Women as farm owners, and participating in the milking, were associated with lower mastitis prevalence on the farm. Peat bedding was associated with higher mastitis prevalence. CONCLUSION: We demonstrated relatively high mastitis prevalence in this study. Contagious bacteria (eg. S. aureus, C. bovis, S. agalactiae and coagulase negative staphylococci) caused most of the infections. These infections are usually spread from cow to cow at milking if the milking hygiene is not good enough. The mastitis situation could be improved by improving milking procedures and hygiene. PMID- 17118213 TI - Proteomics gets faster and smarter. PMID- 17118212 TI - Arthritis and pain. Neurogenic origin of joint pain. AB - Arthritis pain affects millions of people worldwide yet we still have only a limited understanding of what makes our joints ache. This review examines the sensory innervation of diarthroidal joints and discusses the neurophysiological processes that lead to the generation of painful sensation. During inflammation, joint nerves become sensitized to mechanical stimuli through the actions of neuropeptides, eicosanoids, proteinase-activated receptors and ion channel ligands. The contribution of immunocytes to arthritis pain is also reviewed. Finally, the existence of an endogenous analgesic system in joints is considered and the reasons for its inability to control pain are postulated. PMID- 17118214 TI - Vital organ blood flow during high-frequency ventilation. PMID- 17118215 TI - Optimizing neurologic prognosis after cardiac arrest. AB - Neurologic disability is a feared outcome of resuscitation from cardiac arrest. The study by Rech and colleagues in the previous issue of Critical Care describes the use of neuron-specific enolase to inform an early prognosis in patients who survived in-hospital cardiac arrest. In their study 'none of the patients had a DNR order and there was no limitation of life support.' As a result, 10% of patients remained in a vegetative state at 6 months, a higher percentage than in other recent studies. The existence of a population of patients in which all are fully supported without withholding care or withdrawal of care may represent an important research opportunity. High neuron-specific enolase levels have been reported in patients that awoke and seem to occur in studies with a higher percentage of patients in a vegetative state at follow-up (more uniform support). If a comprehensive set of clinical, electrophysiological, biochemical and imaging measures could be obtained in a uniform manner in a cohort of patients without limitations in care, a more objective set of comprehensive prognostic indicators could be obtained. A focused international consortium is called for. PMID- 17118216 TI - Bench-to-bedside review: the role of glycosaminoglycans in respiratory disease. AB - The extracellular matrix (ECM) plays a significant role in the mechanical behaviour of the lung parenchyma. The ECM is composed of a three-dimensional fibre mesh that is filled with various macromolecules, among which are the glycosaminoglycans (GAGs). GAGs are long, linear and highly charged heterogeneous polysaccharides that are composed of a variable number of repeating disaccharide units. There are two main types of GAGs: nonsulphated GAG (hyaluronic acid) and sulphated GAGs (heparan sulphate and heparin, chondroitin sulphate, dermatan sulphate, and keratan sulphate). With the exception of hyaluronic acid, GAGs are usually covalently attached to a protein core, forming an overall structure that is referred to as proteoglycan. In the lungs, GAGs are distributed in the interstitium, in the sub-epithelial tissue and bronchial walls, and in airway secretions. GAGs have important functions in lung ECM: they regulate hydration and water homeostasis; they maintain structure and function; they modulate the inflammatory response; and they influence tissue repair and remodelling. Given the great diversity of GAG structures and the evidence that GAGs may have a protective effect against injury in various respiratory diseases, an understanding of changes in GAG expression that occur in disease may lead to opportunities to develop innovative and selective therapies in the future. PMID- 17118217 TI - Bench-to-bedside review: critical illness-associated cognitive dysfunction- mechanisms, markers, and emerging therapeutics. AB - Cognitive dysfunction is common in critically ill patients, not only during the acute illness but also long after its resolution. A large number of pathophysiologic mechanisms are thought to underlie critical illness-associated cognitive dysfunction, including neuro-transmitter abnormalities and occult diffuse brain injury. Markers that could be used to evaluate the influence of specific mechanisms in individual patients include serum anticholinergic activity, certain brain proteins, and tissue sodium concentration determination via high-resolution three-dimensional magnetic resonance imaging. Although recent therapeutic advances in this area are exciting, they are still too immature to influence patient care. Additional research is needed if we are to understand better the relative contributions of specific mechanisms to the development of critical illness-associated cognitive dysfunction and to determine whether these mechanisms might be amenable to treatment or prevention. PMID- 17118218 TI - Koala retrovirus: a genome invasion in real time. AB - Koalas are currently undergoing a wave of germline infections by the retrovirus KoRV. Study of this phenomenon not only provides an opportunity for understanding the processes regulating retrovirus endogenization but may also be essential to preventing the extinction of the species. PMID- 17118219 TI - Intrauterine exposures, pregnancy estrogens and breast cancer risk: where do we currently stand? AB - Since 1990, when a hypothesis on intrauterine influences on breast cancer risk was published, several studies have provided supportive, indirect evidence by documenting associations of birth weight and other correlates of the prenatal environment with breast cancer risk in offspring. Recent results from a unique cohort of women with documented exposure to diethylstilbestrol in utero have provided direct evidence in support of a potential role of pregnancy oestrogens on breast cancer risk in offspring. PMID- 17118220 TI - The interaction of macrophage receptors with bacterial ligands. AB - Innate immune receptors play a key role in the early recognition of invading bacterial pathogens and initiate the crucial innate immune response. The diverse macrophage receptors recognise Gram-positive and Gram-negative bacteria via conserved structures on the bacterial surface and facilitate phagocytosis and/or signalling, providing the trigger for the adaptive immune response. These receptors include scavenger receptors, C-type lectins, integrins, Toll-like receptors and siglecs. The bacterial ligands generally recognised by these receptors range from lipopolysaccharides on Gram-negative bacteria to peptidoglycan and lipoteichoic acid on Gram-positive bacteria. However, emerging evidence indicates that bacterial proteins are also important ligands; for example, surface proteins from Neisseria meningitidis have been shown to be ligands for class A scavenger receptors. In addition, a group of cytosolic receptors, the NBS-LRR proteins, have been implicated in recognition of bacterial breakdown products. It is becoming increasingly apparent that macrophage receptors can act in conjunction with one another to deliver an appropriate response. PMID- 17118221 TI - An appraisal of aspirin therapy in diabetes. PMID- 17118222 TI - Treatment of prediabetic neuropathy. PMID- 17118223 TI - An update on autonomic neuropathy affecting the gastrointestinal tract. AB - Gastrointestinal symptoms and disordered gut motility occur frequently in the diabetic population and are generally regarded as manifestations of gastrointestinal "autonomic dysfunction," although the relationships between both symptoms and dysmotility with abnormal cardiovascular autonomic function are weak. It is now recognized that the blood glucose concentration is both a determinant of and determined by gastrointestinal function. An improved definition of the underlying pathophysiology should facilitate the development of therapies that are targeted more effectively. PMID- 17118224 TI - Cardiovascular autonomic neuropathy: diagnosis and management. AB - Cardiovascular autonomic neuropathy increases morbidity and mortality, and reduces quality of life and activities of daily living of the patients with diabetes. The reduced cardiovascular autonomic function as measured by heart rate variability is strongly associated with an increased risk of silent myocardial ischemia and mortality. Currently, no specific therapeutic strategies can be recommended for cardiac autonomic neuropathy, but management of hyperglycemia and the use of angiotensin-converting enzyme inhibitors and beta blockers should be instituted. PMID- 17118225 TI - Central nervous system involvement in diabetes mellitus. AB - Diabetic complications result in much morbidity and mortality and considerable consumption of scarce medical resources. Thus, elucidation of the risk factors and pathophysiologic mechanisms underlying diabetic complications is important. The effects of diabetes on the central nervous system (CNS) result in cognitive dysfunction and cerebrovascular disease. Treatment-related hypoglycemia also has CNS consequences. Advances in neuroimaging now provide greater insights into the structural and functional impact of diabetes on the CNS. Greater understanding of CNS involvement could lead to new strategies to prevent or reverse the damage caused by diabetes mellitus. PMID- 17118226 TI - The diabetic foot: the importance of biofilms and wound bed preparation. AB - Biofilms are ubiquitous and medically important complex structures consisting of microbial-associated cells embedded in self-produced extracellular matrix of hydrated extrapolymeric substances, which are irreversibly attached to a biological or nonbiological surface. Bacteria that reside as biofilms are resistant to traditional therapy. This alternative community in which microbes exist has recently attracted interest as a potential reason why chronic wounds do not heal. This may be especially important for diabetic foot ulcers, which are often characterized by their refractory nature, their predisposition to have associated underlying infection, and their improvement with debridement. Animal and in vitro models have been developed to better study biofilms, which will allow a venue for therapeutic intervention. Potential opportunities exist that include prevention of bacterial attachment, prevention of biofilm formation, disruption of the biofilm to allow penetration of topical antimicrobial agents, interference with quorum sensing, and enhancement of bacteria dispersion from biofilms to a more easily destroyed planktonic state. PMID- 17118227 TI - Negative pressure wound therapy: evidence-based treatment for complex diabetic foot wounds. AB - Negative pressure therapy is a novel technology used for the promotion of wound healing and has emerged as the standard care in the management of problem wounds. Negative pressure wound therapy has been met with rapid clinical success and widespread acceptance. The literature is replete with case series, small trials, and noncomparative studies; however, there are few prospective, randomized, human trials examining this technology and its ability to promote healing. We review and evaluate the current literature on negative pressure therapy and its efficacy in the healing of complex diabetic wounds. PMID- 17118228 TI - Clinic versus home blood pressure measurements in patients with CKD. PMID- 17118229 TI - Lipids and diabetic nephropathy. AB - Although several factors may mediate the development and progression of diabetic nephropathy, hyperlipidemia is now considered an independent and major determinant of progression of renal disease in diabetes. The following discussion focuses on the experimental evidence that incriminates hyperlipidemia as a pathogenic factor for diabetic nephropathy and the potential mechanisms that may mediate renal injury from hyperlipidemia, as well as the clinical studies involving therapeutic interventions for hyperlipidemia and their impact on progression of diabetic renal disease. PMID- 17118230 TI - Inflammation and diabetic nephropathy. AB - Diabetic nephropathy has become the main cause of renal failure, but unfortunately the intimate mechanisms leading to the development and progression of renal injury are not yet fully known. Activated innate immunity and inflammation are relevant factors in the pathogenesis of diabetes. Moreover, different inflammatory molecules, including chemokines, adhesion molecules, and proinflammatory cytokines, may be critical factors in the development of microvascular diabetic complications, including nephropathy. This new pathogenic perspective leads to important therapeutic considerations, with new pathogenic pathways becoming important therapeutic targets that can be translated into clinical treatments for diabetic nephropathy. PMID- 17118231 TI - Anemia and diabetic nephropathy. AB - Anemia has recently been recognized as a frequent complication of diabetic nephropathy, appearing earlier than in nondiabetic renal disease and amplifying the risks of cardiovascular and microvascular complications. A major cause is an inappropriate erythropoietin response to anemia, often accompanied by iron deficiency and therapy with angiotensin-converting enzyme inhibitors and angiotensin receptor blockers. PMID- 17118232 TI - Transplantation for type 1 diabetes: whole organ pancreas and islet cells. AB - Beta-cell transplantation can be either a whole organ pancreas transplant or an islet cell transplant; both have resulted in restoration of a normoglycemic state. These transplants can be performed in conjunction with kidney transplants or as solitary grafts. In general, whole organ transplants have more sustained and durable function and are more widely available than their islet counterparts, but entail a much more invasive procedure. Beta-cell transplantation continues to evolve, with considerable improvement in results over the past two decades, and is a cost-effective option for select patients with type 1 diabetes. PMID- 17118233 TI - Diabetic nephropathy: worldwide epidemic and effects of current treatment on natural history. AB - Diabetic nephropathy has become a worldwide epidemic, accounting for approximately one third of all cases of end-stage renal disease. With increasing prevalence of diabetes particularly in Asia, and a global prevalence of microalbuminuria of 39%, the problem is expected to grow. Improved management of diabetes aimed at improved glycemic control, to avoid initiation of diabetic nephropathy, and antihypertensive treatment blocking the renin-angiotensin system, to avoid its progression, need to be implemented, particularly in high risk patients. PMID- 17118235 TI - [Suicidal behavior among young people with eating disorders]. PMID- 17118236 TI - [Suicidal behaviour in a clinical population of 12- to 17-year-old patients with eating disorders]. AB - INTRODUCTION: The aim of this study was to determine the prevalence of suicidal behavior and comorbidity among patients 12-17 years of age with eating disorders. Previous studies showed an increased rate of suicidal behavior in young people with eating disorders. MATERIALS AND METHODS: Data were collected as part of a European study concerning eating disorders. 37 patients (12-17 years, mean age 15 years) from the Center for Eating Disorders, Odense University Hospital, participated in the study. The frequency of suicidal behavior was tested individually by BAB-T interview (''Grading of Anorexia and Bulimia, Teen-ager''), which is a semi-structured interview with the patient, and by YSR (''Youth Self Report'') a questionnaire filled in by the patient herself. RESULTS: Suicidal behavior was shown by 40% of the patients, and 29% had self-injurious behavior. There was a tendency for suicidal behavior and comorbidity to be more prominent in the youngest patients. CONCLUSION: The risk of suicidal behavior is greater in patients with eating disorders than in the background population. In the youngest patients there is an increased tendency to suicidal behavior and comorbidity, but this requires further documentation. The clinician must be aware of self injurious behavior in patients with eating disorders, including those between 12 and 17 years of age. PMID- 17118237 TI - [Cardiac sarcoidosis]. AB - The paper gives a review of the epidemiology, clinical manifestations, diagnosis, treatment and prognosis of cardiac sarcoidosis. Approximately 30% of patients with systemic sarcoidosis have granulomas in the myocardium, and 5% have clinical signs of cardiac affection. Cardiac manifestations comprise pericarditis, heart block, ventricular arrhythmias, valvular disease, ventricular aneurism, congestive heart failure and sudden cardiac death. Most frequent causes of death are sudden death and heart failure. Adequate treatment improves the prognosis of the disease. PMID- 17118238 TI - [Gynaecological epidemiology]. AB - Epidemiological research has good possibilities in Denmark due to the fact that all people have a personal PIN code and due to our many National health registers. In gynaecology the National Register of Patients, the Birth Registry, IVF-registry, Cancer Registry and latest the National Prescription Database offer unique possibilities of linking exposure data with many clinical outcomes. Danish epidemiology has contributed with morbidity analyses on children concieved by in vitro fertilisation, pharmacoepidemiological studies on short and long term effects of oral contraceptives and hormone therapy and, now, with routine assessment of clinical quality. PMID- 17118239 TI - [Quality assurance of antibiotic prescribing. Is it beneficial--and what is most effective?]. PMID- 17118240 TI - [Loss of heterozygosity on chromosomes 1 and 19 in cases of primary brain tumour]. AB - INTRODUCTION: Histological classification of brain tumours, including gliomas, can be difficult, and genetic investigations are increasingly significant in their classification and the development of treatment strategies. Oligodendrogliomas often show a loss of heterozygocity for the short arm of chromosome 1 and the long arm of chromosome 19 (LOH 1p/19q), changes that influence both treatment and prognosis. Our aim was to evaluate the incidence of combined loss of heterozygocity 1p and 19q in various glioma groups. MATERIALS AND METHODS: A total of 10 oligodendrogliomas (5 WHO grade II and 5 grade III), 10 mixed gliomas (5 WHO grade II and 5 grade III), 10 astrocytomas (5 WHO grade II and 5 grade III) and 11 glioblastomas (WHO grade IV) were investigated. Normal hippocampal tissue was used as a control. Formalin-fixed paraffin-embedded tissue was scrutinized with fluorescent in situ hybridization (FISH) with fluorochrome conjugated double-strand DNA probes for 1p and 19q, respectively. RESULTS: A significiant loss of 1p/19q was found in the oligodendrogliomas; the astrocytomas showed a selective loss of 19q; the glioblastomas showed a selective loss of 1p but also polyploidy. CONCLUSION: This investigation confirms other reports on increased LOH 1p/19q in oligodendrogliomas. Various studies have demonstrated a large variation in the incidence of LOH 1p/19q. This might be due to inter- and intraobserver variability in the histological classification. Another factor might be variations in techniques. Most studies have been made on imprints. A standard for the method, including the number of cells counted, the cutoff limit and the statistical variation, is necessary for future studies and clinical use. PMID- 17118241 TI - [Self-management versus conventional management of oral anticoagulant therapy: a randomized controlled trial]. AB - The efficacy of self-managed oral anticoagulant therapy has been addressed in few randomized controlled trials, which have provided inconsistent results. The aim of this study was to compare the quality of self-managed oral anticoagulant therapy with conventional management. One hundred patients were randomly assigned to the two treatment regimens, and it was found that the quality of self management of oral anticoagulant therapy was at least as good as the treatment provided by conventional management. Self-management is therefore a valid management option in selected patients. PMID- 17118242 TI - [Cognitive functions in elderly high-risk patients after off-pump coronary artery bypass grafting versus conventional bypass grafting - a randomised study - Secondary publication]. AB - It has been suggested that the risk of cerebral dysfunction is less with off-pump coronary artery bypass grafting (OPCAB) than with conventional coronary artery bypass grafting (CCAB). The study is a part of the Best Bypass Surgery Trial. A total of 120 elderly patients underwent psychometric testing with a neuropsychological test battery before as well as 3 months after surgery. Cognitive dysfunction was identified in 7.4% of the patients in the OPCAB group and 9.8% in the CCAB group. We found no significant difference after either OPCAB or CCAB. PMID- 17118243 TI - [Cardiac sarcoidosis--a difficult diagnosis. A report of 8 consecutive patients with arrhythmias and cardiomyopathy]. AB - During the period 1984-2005, eight patients aged 25-57 years were treated for cardiac sarcoidosis. Diagnosis was obtained in three patients by endomyocardial biopsy, in three at heart transplantation (HTx) and in two at autopsy. Two patients had heart block, five ventricular arrhythmias and six dilated cardiomyopathy with congestive heart failure. Five patients died, one by sudden death, two of heart failure and two after HTx. Diagnosing cardiac sarcoidosis remains difficult, although MRI and PET scan have renewed the awareness of the disease. PMID- 17118244 TI - [Severe skin rash in patients treated with terbinafine]. AB - We report on five cases of severe skin rash in patients treated with oral terbinafine. Four patients were treated without mycological testing. Two patients were diagnosed with candidiasis on the skin or in the fingernails and were incorrectly treated with terbinafine. PMID- 17118245 TI - [The first HPV vaccine is now available]. AB - More than two thirds of carcinomas of the uterine cervix are caused by human papilloma virus (HPV) types 16 and 18, and 90% of all genital warts are caused by HPV 6 and 11. In June 2006, the US Food and Drug Administration accepted the first prophylactic HPV vaccine against HPV 6, 11, 16 and 18 to be offered to girls and young women. This new vaccine is also now available in Denmark. Questions concerning economics, ethics, organization, and vaccine monitoring need to be discussed in connection with the establishment of a vaccine program. In addition, information to the general population as well as to health care providers and decision-makers should have been given a high priority. PMID- 17118247 TI - [Copy is fraudulent--but what is the consequence?]. PMID- 17118248 TI - [Treatment of potentially life-threatening diseases with laser and intense pulsed light by non-medically trained persons]. PMID- 17118249 TI - [Does the modern office environment desiccate the eyes?]. AB - Eye irritation is a common complaint in the modern office environment. The causes are still unknown. It appears that high preocular relative humidity protects the precorneal tear film against desiccation and airborne chemicals and reduces the development of eye irritation by airborne sensory irritants. This is particularly relevant for intensive computer work, where the precorneal tear film is altered, resulting in dryness of the eye. The reduction of the blink frequency during computer work promotes desiccation of the precorneal tear film; the frequency is moderately correlated with both the tear film thickness and the aqueous loss. For this reason, it is important to plan the workplace and work schedule in such a way to maintain a normal eye blink frequency to minimise alterations of the precorneal tear film. PMID- 17118250 TI - [Cannabis and psychosis]. AB - Longitudinal studies of the association between use of cannabis and later psychosis or schizophrenia were reviewed. Studies included were studies in the general population and in conscripts, using information from interviews about use of cannabis and register-based follow-up in psychiatric case-registers or in personal interviews. There was a consistent finding that use of cannabis was associated with an increased risk of later psychosis with an odds ratio of approximately 2, when adjusted for predisposition to mental illness and socio demographic risk factors. PMID- 17118251 TI - [Treatment by non-physicians of skin diseases--including potentially malignant diseases with lasers and intense pulsed light]. AB - INTRODUCTION: Laser and IPL treatment by non-physicians raises professional concern. The purpose of this investigation is to evaluate laser and IPL treatment carried out by non-physicians and to assess the level of pre-treatment information given to patients. MATERIAL AND METHODS: Approached clinics were found by internet-searches and from advertisements in telephone directories and national newspapers. The target group was clinics in Zealand that offer laser or IPL treatment of pigmented lesions, pigmented nevi, sun-damaged skin, acne and/or unwanted hair growth. The investigation did not include specialised clinics run by dermatologists and plastic surgeons. By means of anonymous telephone calls the clinics were presented for standardized questions under the pretext of being a potential client. RESULTS: Of 28 clinics investigated, 93% offered treatment for unwanted hair growth, 75% for pigmented lesions, 50% for acne, 36% for possible actinic keratoses and 29% for pigmented nevi. Medical examination was an exception (11%). In none of the clinics were medical examinations performed by specialists in dermatology or plastic surgery. Cosmeticians or nurses generally gave the laser and IPL treatments. In 57% of the clinics patients were informed that the treatment did not have any risks. In June treatment was offered in 79% of the clinics, 18% of which mentioned that no special precautions were necessary when treating in sunny periods. CONCLUSION: Laser and IPL treatment of skin diseases, including potential malignant diseases, is carried out by non physicians and pre-treatment information contains major errors and shortcomings. PMID- 17118252 TI - [Skin prick tests of patients with hay fever carried out in general practices compared with those carried out in a specialist outpatient clinic]. AB - INTRODUCTION: The aim of our study was to assess whether general practitioners and their staff (practices) who had attended a short CME course with technical instruction in the skin prick test could diagnose and treat unselected patients with allergic rhinoconjunctivitis at the same quality level as the allergy outpatient clinic. MATERIAL AND METHODS: We performed a multicentre study with the participation of 38 general practices and the Allergy Centre at Odense University Hospital (OUH). After a two-day course for the general practitioner and his practice staff, every practice performed a skin prick test on 10 consecutive adult patients with symptoms of allergic rhinoconjunctivitis during the spring and summer. A standardised questionnaire was also filled in and sent to the Allergy Centre, where the patient subsequently had another skin prick test carried out. The results of the tests were determined in duplicate and then compared. RESULTS: No significant differences in the quality of the skin prick test for 10 allergens or histamine control were found between the general practice and the Allergy Centre. Discordant results were found in 9%, i.e., a positive result either at the practice or the Allergy Centre, but not at both. CONCLUSION: After a training course, general practitioners and their staff are fully able to perform and validate skin prick tests for inhalation allergens. PMID- 17118253 TI - [Myocardial scintigraphy. Clinical use and consequence in a non-invasive cardiological department]. AB - INTRODUCTION: Myocardial perfusion imaging (MPI) is increasingly used for the diagnosis of ischaemic heart disease. The method is particularly applied as a gate keeper before coronary angiography (CAG) in patients with intermediate probability for ischaemic heart disease. This study aimed to analyse the clinical use of MPI in a university hospital without invasive cardiological laboratory. MATERIAL AND METHODS: In the period 01.01.2002 to 31.12.2003, 259 patients (141 women, 118 men) were referred to MPI from our department of cardiology. RESULTS: Normal MPI was seen in 111 patients (43%), whereas reversible ischaemia was seen in 88 patients (34%) and led to referral of 52 patients (59%) to CAG. 17 patients (19%) continued clinical control, and 19 cases (22%) were closed. Correlating results between MPI and all performed CAGs were found in 42 patients (61%), and divergent results were seen in 27 (39%) patients. CONCLUSION: Referral to CAG is primarily based on the combination of the results of MPI and the clinical symptoms of ischaemic heart disease. PMID- 17118254 TI - [Clinical microbiological specimen collection: tissue specimens and culture]. PMID- 17118255 TI - [Weight changes and the effect of anti-diabetes treatment in the first 5 years after diabetes diagnosis - secondary publication]. AB - We monitored changes in patients' weight during the first 5 years after diabetes diagnosis. Data were from 711 newly diagnosed diabetic patients aged 40 or over managed in general practice. Patients whose only treatment was advice on diet generally maintained an initial weight loss of 5-7 kg over the 5 years. Patients receiving metformin or sulfonylureas maintained an average weight loss of 2-4 kg depending on age and sex, while an acceptable average glycaemic control was achieved. The results indicate that weight reduction is a practicable treatment in diabetic patients. PMID- 17118256 TI - [A prospective study of drinking habits and coronary heart disease in middle-aged Danish men and women - secondary publication]. AB - Individuals with a moderate alcohol intake are at lower risk of heart disease than abstainers. In this study, we examined whether the frequency of alcohol intake plays a role in this association. Among men, we observed a low risk of coronary heart disease among frequent drinkers that was independent of the total weekly intake. Among women, we observed a low risk of coronary heart disease that was independent of the drinking frequency. These sex differences in the association of alcohol with coronary heart disease deserve more research. PMID- 17118257 TI - [Avascular necrosis in a man with HIV infection]. AB - Recently, a high incidence of avascular necrosis (AVN) has been reported in HIV infected individuals. We present a further case of AVN of both femoral heads, the left distal femur and the proximal tibia in a 19-year-old HIV-positive man. The patient complained of severe pain in both hips, which had lasted 10 months. The diagnosis was aided by radiological assessment and radioisotope bone scan and confirmed by magnetic resonance imaging. The patient had been receiving antiretroviral therapy for 30 months and had no other known risk factors for developing AVN. We recommend early evaluation of musculoskeletal pain in HIV infected patients to rule out AVN. PMID- 17118258 TI - [Idiopathic brachial neuritis in a 12-year-old boy]. AB - Idiopathic brachial neuritis (IPN) is a clinical entity of unknown cause characterized by sudden onset of pain and weakness with occasional atrophy of the arm muscles. It has rarely been documented in children. Many cases are preceded by upper respiratory tract infections, immunisations or trauma, which has led to speculation about an autoimmune cause. A case of severe IPN in a 12-year-old boy without antecedent events is presented. Diagnostic tests, treatment and prognosis are discussed. PMID- 17118259 TI - Dynamics of arbuscular mycorrhizal symbiosis in heavy metal phytoremediation: meta-analytical and conceptual perspectives. AB - To estimate dynamics of arbuscular mycorrhizal (AM) symbiosis in heavy metal (HM) phytoremediation, we conducted a literature survey and correlated HM uptake and relative plant growth parameters from published data. After estimating AM feedback responses for these parameters at low and high soil-HM concentration intervals, we determined that the roles of AM symbiosis are characterized by (1) an increased HM phytoextraction via mycorrhizospheric 'Enhanced Uptake' at low soil-HM concentrations, and (2) a reduced HM bioavailability via AM fungal 'Metal Binding' processes at high soil-HM levels, hence resulting in increased plant biomass and enhanced plant tolerance through HM stress-avoidance. We present two conceptual models which illustrate the important compromise between plant growth, plant HM uptake and HM tolerance, and further emphasize the importance of AM symbiosis in buffering the soil environment for plants under such stress conditions. PMID- 17118260 TI - Evaluation of the treatment efficiencies of paper mill whitewaters in terms of organic composition and toxicity. AB - The efficiency of several lab scale treatments (aerobic, anaerobic and ozone or combination of these) was evaluated using two packaging board mill whitewaters. The effect of the different treatments on the elimination of the organic load, the chemical oxygen demand (COD) and the toxicity was tested as well as the relationship between these parameters. Biocides, phenolic compounds, surfactants, plasticiziers and wood extractives were identified in untreated and treated whitewaters by liquid chromatography coupled with mass spectrometry (LC-MS) or gas chromatography coupled with mass spectrometry (GC-MS). A strong dependency on the water type and treatment efficiency was observed, being the combination of anaerobic and aerobic treatments the best option to reduce the organic contaminants in these waters, although in some cases, the toxicity did not decrease. However, ozone as post-treatment permitted a further reduction of organic compounds, toxicity and COD. PMID- 17118261 TI - Methodological issues in infertility research. AB - It is now well established that clinical decision-making can be enhanced by incorporating evidence from valid studies. This evidence-based approach to health care management requires a critical appraisal of the available evidence, most of which is of relatively poor quality. If the best evidence is to be put forward to support clinical decisions then it must be derived from studies of high methodological rigour. This chapter discusses important methodological issues affecting the evaluation of the efficacy of therapeutic interventions; these are supplemented with illustrative examples from the literature. These issues form the basis for establishing an approach to inform gynaecological practice, with particular reference to infertility management using assisted reproductive technology and other therapies. Ten important issues are discussed: the formulation of an appropriate research question, the use of randomization, the importance of concealment of treatment allocation, the importance of blinding or masking to avoid ascertainment bias, the avoidance of cointervention, the requirement of an adequate sample size, the restriction of evaluation to the first cycle of treatment, the avoidance of the crossover trial when pregnancy is the outcome of interest, the importance of analysing data using an intention-to treat approach, and the clear identification of the orientation of the study from the perspectives of superiority, equivalence or non-inferiority of the interventions being compared. PMID- 17118262 TI - BMP signaling in the cartilage growth plate. AB - Transforming growth factor-beta (TGF-beta) superfamily members play diverse roles in all aspects of cartilage development and maintenance. It is well established that TGF-betas and bone morphogenetic proteins (BMPs) play distinct roles in the growth plate. This chapter discusses key experiments and experimental approaches that have revealed these roles, and progress toward the identification of previously unsuspected roles. Current understanding of the mechanisms by which different TGF-beta and BMP pathways exert their functions is discussed. Finally attempts to utilize this information to promote cartilage regeneration, and important issues for future research, are outlined. PMID- 17118263 TI - The CLIP-170 orthologue Bik1p and positioning the mitotic spindle in yeast. AB - Bik1p is the yeast Saccharomyces cerevisiae representative of the CLIP-170 family of microtubule plus-end tracking proteins. Bik1p shares a number of similarities with its mammalian counterpart CLIP-170, including an important role in dynein function. However, Bik1p and CLIP-170 differ in several significant ways, including the mechanisms utilized to track microtubule plus ends. In addition to presenting functional comparisons between Bik1p and CLIP-170, we provide sequence analyses that reveal previously unrecognized similarities between Bik1p and its animal counterparts. We examine in detail what is known about the functions of Bik1p and consider the various roles that Bik1p plays in positioning the yeast mitotic spindle. This chapter also highlights several recent findings, including the contribution of Bik1p to the yeast mating pathway. PMID- 17118264 TI - Aggregate-prone proteins are cleared from the cytosol by autophagy: therapeutic implications. AB - Intracellular protein misfolding/aggregation are features of many late-onset neurodegenerative diseases, called proteinopathies. These include Alzheimer's disease, Parkinson's disease, tauopathies, and polyglutamine expansion diseases [e.g., Huntington's disease; and various spinocerebellar ataxias (SCAs), like SCA3]. There are no effective strategies to slow or prevent the neurodegeneration resulting from these diseases in humans. The mutations causing many proteinopathies (e.g., polyglutamine diseases and tauopathies) confer novel toxic functions on the specific protein, and disease severity frequently correlates with the expression levels of the protein. Thus, the factors regulating the synthesis and clearance of these aggregate-prone proteins are putative therapeutic targets. The proteasome and autophagy-lysosomal pathways are the major routes for mutant huntingtin fragment clearance. While the narrow proteasome barrel precludes entry of oligomers/aggregates of mutant huntingtin (or other aggregate-prone intracellular proteins), such substrates can be degraded by macroautophagy (which we will call autophagy). We showed that the autophagy inducer rapamycin reduced the levels of soluble and aggregated huntingtin and attenuated its toxicity in cells, and in transgenic Drosophila and mouse models. We extended the range of intracellular proteinopathy substrates that are cleared by autophagy to a wide range of other targets, including proteins mutated in certain SCAs, forms of alpha-synuclein mutated in familial forms of Parkinson's disease, and tau mutants that cause frontotemporal dementia/tauopathy. In this chapter, we consider the therapeutic potential of autophagy upregulation for various proteinopathies, and describe how this strategy may act both by removing the primary toxin (the misfolded/aggregate prone protein) and by reducing susceptibility to apoptotic insults. PMID- 17118265 TI - Wnt signaling: a key regulator of bone mass. AB - The identification of a link between bone mass in humans and gain- [high bone mass (HBM) trait] or loss-of-function [osteoporosis pseudoglioma (OPPG) syndrome] mutations in the Wnt coreceptor lipoprotein receptor-related protein (LRP)5 or in the Wnt antagonist sclerostin (sclerosteosis, Van Buchem syndrome) has called the attention of academic and industry scientists and clinicians to the importance of this signaling pathway in skeletal biology and disease. Multiple genetic and pharmacological manipulations of Wnt signaling in mice have since then confirmed the central role of this pathway in both the establishment of peak bone mass and its maintenance throughout life. Wnt signaling appears to be located downstream of bone morphogenetic proteins (BMPs), itself induced by Hedgehog (Hh) signaling, suggesting that it is the successive recruitment of these three intracellular signaling cascades that allow the full expression of the genetic patterns that characterize the osteoblast, the cell responsible for the formation of bone. PMID- 17118266 TI - Eukaryotic DNA replication in a chromatin context. AB - There has been remarkable progress in the last 20 years in defining the molecular mechanisms that regulate initiation of DNA synthesis in eukaryotic cells. Replication origins in the DNA nucleate the ordered assembly of protein factors to form a prereplication complex (preRC) that is poised for DNA synthesis. Transition of the preRC to an active initiation complex is regulated by cyclin dependent kinases and other signaling molecules, which promote further protein assembly and activate the mini chromosome maintenance helicase. We will review these mechanisms and describe the state of knowledge about the proteins involved. However, we will also consider an additional layer of complexity. The DNA in the cell is packaged with histone proteins into chromatin. Chromatin structure provides an additional layer of heritable information with associated epigenetic modifications. Thus, we will begin by describing chromatin structure, and how the cell generally controls access to the DNA. Access to the DNA requires active chromatin remodeling, specific histone modifications, and regulated histone deposition. Studies in transcription have revealed a variety of mechanisms that regulate DNA access, and some of these are likely to be shared with DNA replication. We will briefly describe heterochromatin as a model for an epigenetically inherited chromatin state. Next, we will describe the mechanisms of replication initiation and how these are affected by constraints of chromatin. Finally, chromatin must be reassembled with appropriate modifications following passage of the replication fork, and our third major topic will be the reassembly of chromatin and its associated epigenetic marks. Thus, in this chapter, we seek to bring together the studies of replication initiation and the studies of chromatin into a single holistic narrative. PMID- 17118267 TI - The regulatory network controlling the proliferation-meiotic entry decision in the Caenorhabditis elegans germ line. AB - The germ line of sexually reproducing animals, at some point in development, consists of both proliferating and differentiating cells. Proliferation is needed to increase cell number, ensuring that a sufficient quantity of gametes is produced. Meiotic development is needed to produce gametes that can support embryogenesis, each with half the ploidy of the somatic cells. For the reproductive strategy of a given species, regulating the timing and number of gametes, and thus controlling the timing of differentiation and the extent of proliferation, is very important for reproductive fitness. Therefore, animals have evolved regulatory mechanisms that tightly control and balance the proliferation-initiation of meiotic development (meiotic entry) decision. Genetic analysis has identified signaling mechanisms involved in controlling this balance in some animals, including mice, Drosophila, and Caenorhabditis elegans. In this chapter, we present our understanding of the genetic hierarchy controlling the proliferation-meiotic entry decision in C. elegans. A core regulatory network controls the decision under all known conditions (developmental stage, sex, and growth temperature). It consists of a canonical Notch signaling pathway promoting proliferation by inhibiting two redundant mRNA regulatory pathways, the GLD-1 and GLD-2 pathways, which promote meiotic entry. Superimposed on the core network is a complex set of factors, some yet to be identified, and many with regulatory relationships still poorly understood, which control the activities of the GLD-1 and GLD-2 pathways and possibly parallel pathways. Some of the complexity arises from these regulators acting only under certain conditions. We also highlight major areas where we lack knowledge. For example, it is unknown if the entire population of proliferating cells are stem cells capable of self-renewal or if only a small portion are stem cells and the rest are transit amplifying cells. PMID- 17118268 TI - Regulation of angiogenesis by hypoxia and hypoxia-inducible factors. AB - Maintenance of oxygen homeostasis is critical for the survival of multicellular organs. As a result, both invertebrates and vertebrates have developed highly specialized mechanisms to sense changes in oxygen levels and to mount adequate cellular and systemic responses to these changes. Hypoxia, or low oxygen tension, occurs in physiological situations such as during embryonic development, as well as in pathological conditions such as ischemia, wound healing, and cancer. A primary effector of the adaptive response to hypoxia in mammals is the hypoxia inducible factor (HIF) family of transcription regulators. These proteins activate the expression of a broad range of genes that mediate many of the responses to decreased oxygen concentration, including enhanced glucose uptake, increased red blood cell production, and the formation of new blood vessels via angiogenesis. This latter process is dynamic and results in the establishment of a mature vascular system that is indispensable for proper delivery of oxygen and nutrients to all cells in both normal tissue and hypoxic regions. Angiogenesis is essential for normal development and neoplastic disease as tumors must develop mechanisms to stimulate vascularization to meet increasing metabolic demands. The link between hypoxia and the regulation of angiogenesis is an area of intense research and the molecular details of this connection are still being elaborated. This chapter will provide an overview of current knowledge and highlight new insights into the importance of HIF and hypoxia in angiogenesis in both physiological and pathophysiological conditions. PMID- 17118269 TI - Posttranslational nitrotyrosination of alpha-tubulin induces cell cycle arrest and inhibits proliferation of vascular smooth muscle cells. AB - Hyperproliferation of vascular smooth muscle cells is a hallmark of atherosclerosis and related vascular complications. Microtubules are important for many aspects of mammalian cell responses including growth, migration and signaling. alpha-Tubulin, a component of the microtubule cytoskeleton, is unique amongst cellular proteins in that it undergoes a reversible posttranslational modification whereby the C-terminal tyrosine residue is removed (Glu-tubulin) and re-added (Tyr-tubulin). Whereas the reversible detyrosination/tyrosination cycle of alpha-tubulin has been implicated in regulating various aspects of cell biology, the precise function of this posttranslational modification has remained poorly characterized. Herein, we provide evidence suggesting that alpha-tubulin detyrosination is a required event in the proliferation of vascular smooth muscle cells. Proliferation of rat aortic smooth muscle cells in response to serum was temporally associated with the detyrosination of alpha-tubulin, but not acetylation of alpha-tubulin; Glu-tubulin reached maximal levels between 12 and 18h following cell cycle initiation. Inclusion of 3-nitro-l-tyrosine (NO(2)Tyr) in the culture medium resulted in the selective nitrotyrosination of alpha tubulin, that was paralleled by decreased elaboration of Glu-tubulin, decreased expression of cyclins A and E, decreased association of the microtubule plus-end binding protein EB1, and inhibited cell proliferation. Nitrotyrosination of alpha tubulin did not induce necrotic or apoptotic death of rat aortic smooth muscle cells, but instead led to cell cycle arrest at the G(1)/S boundary coincident with decreased DNA synthesis. Collectively, these results suggest that the C terminus of alpha-tubulin and its detyrosination are functionally important as a molecular switch that regulates cell cycle progression in vascular smooth muscle cells. PMID- 17118271 TI - Normal sleep and circadian rhythms: neurobiologic mechanisms underlying sleep and wakefulness. AB - Sleep is a vital, highly organized process regulated by complex systems of neuronal networks and neurotransmitters. Sleep plays an important role in the regulation of central nervous system and body physiologic functions. Sleep architecture changes with age and is easily susceptible to external and internal disruption. Reduction or disruption of sleep can affect numerous functions varying from thermoregulation to learning and memory during the waking state. PMID- 17118272 TI - Insomnia: prevalence, impact, pathogenesis, differential diagnosis, and evaluation. AB - When patients report problems sleeping, a psychiatrist must determine their significance based on frequency, duration, and daytime impairment. Because up to 50% of adults report sleep problems in any year, it is necessary to define when insomnia becomes long-standing, severe, and a complication to daytime function. Psychiatrists must determine if a sleep disturbance reduces mood, motor performance, or cognitive function. If insomnia syndrome is present, major depression, dysthymia, and anxiety disorders commonly are comorbid. To assist in evaluating insomnia, psychiatrists are urged to use the 6 Ps + M of insomnia model to conceptualize the characteristics of the insomnia and coordinate therapeutic intervention. PMID- 17118273 TI - Pharmacologic management of insomnia: past, present, and future. AB - Views on the etiology and morbidities associated with insomnia are evolving and affect clinicians' approach to the pharmacologic management of insomnia. Currently, benzodiazepine receptor agonists (BzRAs) and a single melatonin receptor agonist are recognized as safe and efficacious hypnotics. Variability in BzRAs, pharmacokinetics, and manipulation of dose provide clinicians with options that meet the needs of most patients. Other drugs (eg, sedating antidepressants) also are used commonly in clinical practice to treat insomnia, but evidence is lacking to support this most cases. Improvement in managing insomnia will result from systematic research with these drugs, with drugs in development, and with novel uses, such as co-therapy. PMID- 17118274 TI - Nonpharmacologic strategies in the management of insomnia. AB - Many psychiatric patients have significant sleep disturbance. Insomnia should be addressed directly even when comorbid with a psychiatric disorder. Nonpharmacologic treatments are effective and especially well suited for long term management of sleep problems. Although the techniques themselves are fairly straightforward, they work best when applied with the kind of clinical insight and experience that psychiatrists regularly draw on in their practices. This article briefly reviews the evaluation of insomnia, with the aim of eliciting clinical material sufficiently comprehensive to inform the choice of treatment, and provides a practical overview of the basic nondrug approaches to insomnia, emphasizing what the clinician and the patient may expect from their application. PMID- 17118275 TI - Excessive daytime sleepiness: considerations for the psychiatrist. AB - Excessive daytime sleepiness or pathologic sleepiness is a complaint found in patients who experience somnolence at unwanted times and adversely affects their daytime function. Although psychiatric illness, chronic medical illness, or medication side effects may be causes for fatigue, insufficient sleep is the most common cause of excessive daytime sleepiness in the general population. When an individual complains of frank sleepiness, in addition to insufficient sleep, important considerations in these patients are disturbances in the normal homeostatic mechanisms that govern sleep and wakefulness. This article summarizes the clinical presentation, the differential diagnosis, commonly used diagnostic tools, and treatment options for patients complaining of excessive daytime sleepiness. PMID- 17118276 TI - Selected sleep disorders: restless legs syndrome and periodic limb movement disorder, sleep apnea syndrome, and narcolepsy. AB - Sleep disorders, including restless legs syndrome and periodic limb movement disorder, sleep apnea syndrome, and narcolepsy, are prevalent medical conditions, likely to be seen by practicing psychiatrists. Awareness of these conditions and their presentations, pathophysiology, and treatment allows psychiatrists to treat these conditions where appropriate, to minimize complications and health consequences associated with delayed diagnosis, and to reduce the burden of disease that these conditions may place on patients already experiencing primary psychiatric disorders. PMID- 17118277 TI - Parasomnias. AB - Parasomnias are undesirable behaviors that arise from sleep but are not fully under voluntary control. Parasomnias are grouped broadly according to whether they arise from non-rapid eye movement (NREM) or rapid eye movement (REM) sleep. NREM parasomnias are disorders of arousal that occur along a continuum of behavioral, affective, and autonomic activation. REM-related parasomnias include REM sleep behavior disorder, sleep paralysis, and nightmare disorder. Parasomnias can often be managed successfully using behavioral and pharmacologic therapies. PMID- 17118278 TI - Circadian rhythm sleep disorders and phototherapy. AB - Circadian rhythm sleep disorders are characterized by a desynchronization between the timing of the intrinsic circadian clock and the extrinsic light-dark and social/activity cycles resulting in symptoms of excessive sleepiness and insomnia. This article explores the six recognized circadian rhythm sleep disorders: delayed sleep phase syndrome, advanced sleep phase syndrome, non-24 hour sleep-wake syndrome, irregular sleep-wake pattern, shift work sleep syndrome, and time zone change syndrome. Additionally discussed are the therapeutic roles of synchronizing agents, such as light and melatonin. PMID- 17118279 TI - Sleep in mood disorders. AB - Sleep disturbances are among the most common symptoms in patients who have acute episodes of mood disorders, and patients who have mood disorders exhibit higher rates of sleep disturbances than the general population, even during periods of remission. Insomnia and hypersomnia are associated with an increased risk for the development or recurrence of mood disorders and increased severity of psychiatric symptoms. Sleep electroencephalogram recordings have identified objective abnormalities associated with mood disorders, providing insight into the neurobiologic relationships between mood and sleep. Future studies will continue to investigate this association and potentially improve treatment of sleep and mood disorders. PMID- 17118280 TI - Sleep in schizophrenia: impairments, correlates, and treatment. AB - In untreated schizophrenia, psychotic decompensation is associated with profound insomnia, one of the prodromal symptoms associated with psychotic relapse. First- and second-generation antipsychotic medication can ameliorate this insomnia, but side effects may include sedation or residual insomnia. Patients who are clinically stable and medicated may continue to experience disturbed sleep, including long sleep-onset latencies, poor sleep efficiency, slow wave sleep deficits, and short rapid eye movement latencies. Schizophrenia also can be associated with comorbid sleep disorders, which may be enhanced or induced by antipsychotic medication. Sleep disorders in schizophrenia should be treated vigorously because normalized sleep and its restorative processes may be essential for a positive clinical outcome. PMID- 17118281 TI - Sleep and anxiety disorders. AB - Sleep disturbances commonly are associated with anxiety disorders, in particular generalized anxiety disorder, panic disorder, and posttraumatic stress disorder. Sleep loss may exacerbate and contribute to relapse of these conditions. Core features of panic disorder and posttraumatic stress disorder occur in relation to sleep (sleep panic attacks or re-experiencing nightmares). Investigation of sleep in anxiety disorders provides clues to mechanisms of arousal regulation relevant to insomnia and pathologic anxiety. Established treatments for anxiety disorders and insomnia have many overlapping components; however, optimal sequencing and integration of the approaches remain under-investigated. PMID- 17118282 TI - Sleep and sleep disorders in children and adolescents. AB - Pediatric sleep disorders are common, affecting approximately 25% to 40% of children and adolescents. Although there are several different types of sleep disorders that affect youth, each disorder can have a significant impact on daytime functioning and development, including learning, growth, behavior, and emotion regulation. Researchers are only beginning to uncover the interaction between sleep and psychiatric disorders in children and adolescents, including depression, attention-deficit/hyperactivity disorder, and autism. This article reviews normal sleep and sleep disorders in children and adolescents, the assessment of sleep in pediatric populations, common pediatric sleep disorders, and sleep in children who have common psychiatric disorders. PMID- 17118283 TI - Sleep and its disorders in older adults. AB - For many older adults, aging is associated with significant changes in sleep. There are a variety of potential causes, including primary sleep disorders, circadian rhythm disturbances, insomnia, depression, medical illness, and medications. As with younger adults, the diagnosis requires a thorough sleep history and an overnight sleep recording when appropriate. Treatment should address the primary sleep problem and can result in significant improvement in quality of life and daytime functioning in older adults. PMID- 17118284 TI - Sleep disorders in women: clinical evidence and treatment strategies. AB - Sleep disorders are more prevalent in women than in men. Sex hormones modulate sleep-wake behaviors and mood and may contribute to heightened risk across the life cycle of women. Sleep disorders may have a unique expression in women, emerging throughout their reproductive life cycle. These conditions require careful treatment strategy to manage medical, hormonal, and behavioral contributing factors to poor sleep efficiency and impaired quality of life. This review focuses on clinical evidence for sleep disorders in women and discusses existing evidence of risk factors and treatment options for insomnia and sleep disordered breathing in women. PMID- 17118285 TI - Sleep and psychiatric disorders: future directions. AB - Understanding of the relationship between co-occurring sleep and psychiatric disorders has undergone a radical change. The longstanding perspective that sleep problems invariably are a symptom of a psychiatric disorder is giving way to understanding that complex bidirectional relationships may exist. This change has opened doors to new directions in research and led to changes in guidelines for clinical practice. This article discusses promising future directions for building on this foundation, including developing lines of research currently underway, studying mechanisms that underlie the relationships between sleep and psychiatric disorders; and developing treatment strategies that target these mechanisms to lead to better treatment of sleep disorders and psychiatric disorders. PMID- 17118288 TI - Clinical applications of the polymerase chain reaction: an update. AB - The development, in the past decade, of nucleic acid amplification and detection methods is useful in the study of the etiopathogenesis, diagnosis, and management of a variety of clinical (including rheumatologic) disorders. An association between infectious agents and rheumatic disorders has been established through such methods as polymerase chain reaction. This article describes the principles behind polymerase chain reaction-based diagnosis and updates its clinical applications. It is beyond the scope of this article, however, to describe other nucleic acid amplification methods or to include a complete list of all polymerase chain reaction assays that have been developed. Other recent reviews offer additional details. PMID- 17118289 TI - Skin and soft tissue infections. AB - Primary skin infections (ie, pyodermas) typically are initiated by some breach in the epidermis, resulting in infection by organisms, such as Streptococcus pyogenes and Staphylococcus aureus, that normally colonize the skin. Host associated factors, such as immunosuppression, vasculopathy, neuropathy, or decreased lymphatic drainage, may predispose to skin infection. The clinical syndromes associated with skin infections are often characteristic and are defined most simplistically by anatomic distribution. Although often mild and self-limited, skin infections can be more aggressive and involve deeper structures, including fascia and muscle. This article discusses skin and soft tissue infections, including impetigo, hair follicle-associated infections (ie, folliculitis, furuncles, and carbuncles) erysipelas, cellulitis, necrotizing fasciitis, pyomyositis, septic bursitis, and tenosynovitis. PMID- 17118290 TI - Advances in the management of septic arthritis. AB - Septic arthritis still continues to be a common and serious problem at major urban medical centers and is one of the most rapidly destructive forms of acute arthritis. The yearly incidence of bacterial arthritis varies from 2 to 10 per 100,000 in the general population to 30 to 70 per 100,000 in patients with rheumatoid arthritis and in patients with joint prostheses. Irreversible loss of joint function may develop in up to 50% of the patients. Despite better antimicrobial agents and improved hospital care, the fatality rate for this medical problem has not changed substantially during the past 30 years. An understanding of the risk factors and the pathogenesis of nongonoccocal bacterial arthritis and other forms of infectious arthritis, primarily in the context of a differential diagnosis and treatment, are important to avoid the delay in making a correct diagnosis and to improve the prognosis. PMID- 17118291 TI - Imaging of osteomyelitis: current concepts. AB - Osteomyelitis frequently requires more than one imaging technique for an accurate diagnosis. Conventional radiography still remains the first imaging modality. MRI and nuclear medicine are the most sensitive and specific methods for the detection of osteomyelitis. MRI provides more accurate information regarding the extent of the infectious process. Ultrasound represents a noninvasive method to evaluate the involved soft tissues and cortical bone and may provide guidance for diagnostic or therapeutic aspiration, drainage, or tissue biopsy. CT scan can be a useful method to detect early osseous erosion and to document the presence of sequestra. PET and SPECT are highly accurate techniques for the evaluation of chronic osteomyelitis, allowing differentiation from soft tissue infection. PMID- 17118292 TI - Reactive arthritis: defined etiologies, emerging pathophysiology, and unresolved treatment. AB - ReA is unique in that it is one of the few disease states of which there is a known trigger. This insight into disease initiation has led to great advances in the pathophysiology. Despite this detailed knowledge, the proper treatment remains elusive. In the years to come it is possible that the specific treatment will be dictated by the triggering microbe. PMID- 17118293 TI - Infections in systemic connective tissue diseases: systemic lupus erythematosus, scleroderma, and polymyositis/dermatomyositis. AB - In SLE, scleroderma, and PM/DM, infections are important causes of morbidity and mortality. This increased risk for developing infections is the result of immune abnormalities and of organ system manifestations associated with these diseases and their treatments. Common bacteria are responsible for most mild and lethal infections; however, opportunistic microorganisms cause death in some patients, particularly in those receiving high doses of corticosteroid and immunosuppressive therapy. Various viral and fungal infections also contribute to the morbidity and mortality associated with these diseases. Regardless of the cause of infections, adequate and prompt recognition and proper treatment of the infected patient are imperative. Patients who have these diseases, especially when receiving high doses of corticosteroids and immunosuppressive therapy, need to be monitored closely for these infections. This care and concern is necessary to ensure optimal patient outcomes, both in morbidity and mortality. PMID- 17118294 TI - Hepatitis C and arthritis: an update. AB - Extrahepatic symptoms during chronic hepatitis C virus (HCV) infection are common and varied. Arthritis can be seen either as part of autoimmune processes (eg, associated with cryoglobulinemia) or independently. Whether the manifestation is specifically attributable to HCV infection or rather to the nonspecific result of a chronic inflammatory process is not clear. The literature available at this time is insufficient to guide the most appropriate course of treatment of HCV arthritis. Standard antirheumatic treatment can be considered, but with caution, because some of these medications occasionally may be hepatotoxic and response to therapy seems variable. Treatment decisions should be determined on a case-by case basis. PMID- 17118295 TI - Rheumatic manifestations of human immunodeficiency virus infection. AB - Rheumatic complaints are common in patients with HIV, and HIV positivity confers an increased susceptibility in populations with similar risk factors for HIV infection. With the advent of the modern combined antiretroviral treatment, HAART has had a profound beneficial effect on survival in HIV-infected patients, with lifelong control of HIV infection and normalization of life expectancy; but it has also contributed to both an altered frequency and a different nature of rheumatic complications now being observed in this population, with new rheumatic complications, such as osteoporosis, osteonecrosis, gout, mycobacterial, mycotic osteoarticular infections, and neoplasia perhaps more prevalent. Rheumatologists, internists, and general physicians need to be aware of these changes to provide optimal diagnosis and how to disclose the results to their patients. They also need to be familiar with the management of HIV infection and to direct careful attention to the prevention of HIV transmission in health care facilities. PMID- 17118296 TI - Role of endogenous retroviruses in autoimmune diseases. AB - Molecular epidemiologic proof that HERVs and other retroelements are involved in autoimmunity or other disorders is complicated by their large numbers in the human genome. As discussed, most HERVs are no longer functional or active because of the accumulation of mutations, frameshifts, and deletions. Detection or quantification of HERV transcripts that may be pathologically involved in a particular autoimmune disease thus is often compromised by the presence in great excess of related, but nonfunctional, RNA. This phenomenon should not deter active work in the field, although it will require development of improved methods to discriminate accurately between closely related RNA transcripts. Development of improved immunologic methods to precisely identify epitopes on autoantigens or rare self-reactive T-cell clones may further implicate HERVs and the other repetitive elements in regulation of the immune system in health and disease. PMID- 17118297 TI - Impact of biologic agents on infectious diseases. AB - Until recently, inflammatory diseases, collagen vascular diseases, inflammatory bowel diseases, and multiple sclerosis were met with a limited offering for treatment. The introduction of biologic agents has revolutionized the approach to these diseases, offering many patients freedom from disease activity staving off resultant destruction to organs and joints with marked improvement in quality of life and disability. This article focuses on the development of serious infections associated with the use of biologic agents. Presented is a synthesis of case series, reports, and systematic reviews to elucidate implicated pathogens and clinical presentations in patients being treated with biologic agents and to form a cursory backbone for prevention and treatment strategies to which clinicians prescribing these agents or encountering patients already on these agents can readily refer. Maintenance of a high index of suspicion is imperative for the prevention and appropriate treatment of serious life-threatening infections in these patients. PMID- 17118299 TI - Implementing best practice strategies to prevent infection in the ICU. AB - Critically ill patients are at high risk for infection because of many factors. Quality and regulatory groups have published guidelines regarding infection prevention in the ICU. A multifaceted, multiprofessional team approach is necessary to develop and implement strategies to prevent infection in the critically ill patient. Bundles of intervention along with daily rounds an assessment are essential program components. Ongoing surveillance and re- education and reinforcement are also part of a strong infection prevention program. PMID- 17118300 TI - Ventilator-associated pneumonia and pressure ulcer prevention as targets for quality improvement in the ICU. AB - The health care culture must change. Florence Nightingale wrote [8] "deep-rooted and universal is the conviction that to give a medicine is to be doing something, or rather everything and to give air, warmth, cleanliness etc. is to do nothing." Hygiene care practices and mobility activities are fundamental and independent care components in the nursing profession. When implemented using available evidence, they can significantly improve patient outcomes. It is time to claim and demonstrate the importance of consistent delivery of the fundamentals of basic nursing care. Interventional patient hygiene is an effective framework to ensure the the basics of nursing care are consistently applied to improve patient outcomes. PMID- 17118301 TI - Developing and implementing quality initiatives in the ICU: strategies and outcomes. AB - The Institute of Medicine identifies quality health care as care that is safe, time, effective, efficient, equitable, and patient centered. In the ICU, where patients who have complex, high-acuity are at increased risk of complications, morbidity, and mortality, promoting quality-focused case is especially important. This article describes several performance-improvement initiatives that were developed and implemented a Midwestern community hospital during a 4-year period. The initiatives to reduce catheter-associated blood stream infections, to identify early sepsis, and to promote evidence-based care. PMID- 17118302 TI - Nursing lessons from the MHA keystone ICU project: developing and implementing an innovative approach to patient safety. AB - Given the pivotal role of nurses in providing and supervising patient care, it is essential that nursing professionals are engaged fully in making care safer. Nursing involvement was instrumental in the Michigan Health and Hospital Association Keystone ICU Project, which resulted in rapid reduction in catheter related blood stream infection rates and ventilator-associated pneumonia rates. Nurses of every credential and every nursing position participated in this broad scale improvement effort. This article describes the MHA Keystone ICU Project, including challenges implicit in changing nursing practice and team behavior in the ICU. The improvement strategies implemented by Keystone ICU teams, and lessons learned by nurses engaged in the work, are likely to have application in other clinical settings. PMID- 17118303 TI - Implementing a rapid response team: factors influencing success. AB - Rapid response teams (RRTs), or medical emergency teams, focus on preventing a patient crisis by addressing changes in patient status before a cardiopulmonary arrest occurs. Responding to acute changes, RRTs and medical emergency teams are similar to "code" teams. The exception, however is that they step into action before a patient arrests. Although RRTs are acknowledge as an important initiative, implementation can present many challenges. This article reports on the implementation and ongoing use of a RRT at a community health care setting, highlighting important considerations and strategies for success. PMID- 17118304 TI - Medication reconciliation: developing and implementing a program. AB - During the past 5 years since the medication reconciliation process was formalized and automated, it has become an independent redundancy. The patient intervention rates are maintained at 30% to 35%, with ADE rates related to medication reconciliation at zero. The medication process takes into account the accuracy and appropriateness of restarting prehospital medications and current ICU medications. It includes the omission of important home medications along with inaccuracies of dosages and frequencies. This form assures that the patient is receiving continuity of care ad decreases complications of the patients health related to the changing of medications. Until recently this concept was disseminated by the staff without consistent administrative support. It was a process developed by nurses and perpetuated by nurses. Recently the administration has mandated that the process be implemented throughout the institution. A Hopkins health care-based collaborative is working to implement medication reconciliation hospital wide. The challenge exists in standardizing a process that is now specific to each functional unit. Multidisciplinary monthly meetings provided a forum for working through the barriers to incorporate these changes. This low-cost, high-impact safely initiative, if planned and performed strategically, can have a significant effect on patient safety. PMID- 17118305 TI - Targeting errors in the ICU: use of a national database. AB - The authors believe that as we move from viewing adverse event reporting system as punitive, and as the safety culture improves, reporting will likely increase. Voluntary incident reporting systems can be used to improve patient safety in the ICU by identifying broken or inadequate systems that lead to adverse events [26]. Voluntary external reporting systems such as the ICUSRS can be used to target errors and produce evidence-based best practice measures to improve patient safety in the ICU. PMID- 17118306 TI - Standardizing IV infusion medication concentrations to reduce variability in medication errors. AB - Practical strategies for preventing medication errors in pediatric patients are needed. Medication safety can be improved by assessing current practices, developing evidence-based interventions to improve such practices, evaluating the impact of new evidence-based innovations, and providing feedback to clinicians [20]. Nurses at the point of care are well positioned to identify and implement structures and processes to address medication errors so that the most preventable errors become a thing of the past. PMID- 17118307 TI - Evaluation of in-patient interventional cardiology. AB - The ACNP service in this study decreased the TA, TC, and LOS for patients transferred from outlying hospitals for cardiac catheterization or PCI. Patients on the ACNP service were provided prescription for appropriate discharge medications including beta-blockers, aspirin, ACE inhibitors, and lipid-lowering agents more often than patients on the housestaff service. Other aspects of care, including follow-up appointments and elements of patient education, were documented more often for patients on the ACNP interventional cardiac service and expand the role of ACNPs into other areas of acute-care cardiology practice. PMID- 17118308 TI - Implementing an intensive glucose management initiative: strategies for success. AB - Glycemic control has become a hallmark for this ICU. Collectively, staff and hospital leaders changed a culture of casual attitudes toward chaotic glycemic control to an evidence-based atmosphere of decreased mortality, which has received local and national recognition. Implementation of a protocol , which seemed radical at the time, showed significant patient outcomes. Characterized by teamwork and education, the process was developed, initiated and continually evaluated for improvement, with a cornerstone of patient safety. The rich database allows for continual analysis of outcomes. PMID- 17118309 TI - A longitudinal study of advanced practice nursing in singapore. AB - This longitudinal study of the first cohort of APN interns provides information on aspects of the practice and on the development of their role from Singapore perspective. The APN role is a new one in Singapore, providing an opportune time to pioneer and define advanced practice nursing for the future. Longitudinal surveys will be conducted annually among this first cohort for 5 years to identify changes in role development over time. These results will be compared against those obtained from subsequent cohorts of APN interns in Singapore. Continued research into the role of the APN is needed to enhance role acceptance and development, and to ensure that the benefits of having APNs are attained. PMID- 17118310 TI - Perioperative high-dose-rate brachytherapy in locally advanced and recurrent gynecologic cancer: Initial results of a phase II trial. AB - BACKGROUND: This study was undertaken to determine the feasibility of perioperative high-dose-rate brachytherapy (PHDRB) as an adjunct to salvage surgery in primary advanced or recurrent gynecologic cancer. METHODS: Twenty-five patients with either locally advanced (n = 4) or recurrent (n = 21) gynecologic cancer suitable for salvage surgery were included. Unirradiated patients were treated with preoperative chemoradiation followed by salvage surgery and PHDRB (R0 and R1 resections receiving 16 or 24 Gy, respectively). Previously irradiated patients were treated with salvage surgery and PHDRB alone with 32 or 40 Gy for R0 or R1 resections, respectively. RESULTS: Resections were categorized as R0 in 9 patients (36.0%) and R1 in 16 (64.0%). Four previously irradiated patients suffered fatal pelvic bleeding between 8 and 13 months after surgery and PHDRB. After a median follow-up of 20 months (3-55+), the 4-year actuarial local and pelvic controls were 88.1% and 80.8%, respectively. The 4-year distant metastases free survival was 40.9%. Four-year actuarial overall survival was 34.0%, with a median survival of 27.1 months (95% confidence interval: 17.5-36.8). CONCLUSIONS: Local and pelvic control results are excellent for this very high-risk-disease population. PHDRB dose in previously irradiated patients has been shifted to the closest lower level due to unacceptable vascular toxicity. PMID- 17118311 TI - Individual fraction optimization vs. first fraction optimization for multichannel applicator vaginal cuff high-dose-rate brachytherapy. AB - PURPOSE: To challenge the assumption of unchanged interfractional geometry changes in the course of fractionated multichannel vaginal cuff high-dose-rate brachytherapy. METHODS AND MATERIALS: Two methods of treatment planning for delivery of vaginal cuff brachytherapy were compared in 44 applications. Individual fraction optimization (IFO)-performed for the specific geometry of each individual fraction-was compared to first fraction optimization (FFO)-an optimized first fraction, applied unaltered for geometry of subsequent fractions in the same patient. Dose difference to critical organs was expressed as the percentage of the prescribed dose. RESULTS: In the paired analysis for IFO vs. FFO, mean and maximum rectal and bladder doses were similar. However for FFO, an excess of greater than 20% mean dose to either bladder or rectum was observed in 41% of cases. Maximum organ doses were exceeded by 20% in 54.5% of applications. CONCLUSIONS: On the basis of these findings, it can be concluded that IFO may be important to minimize doses to critical structures. PMID- 17118312 TI - To plan or not to plan? That is the question. PMID- 17118313 TI - High-dose-rate interstitial brachytherapy for gynecologic malignancies. AB - PURPOSE: The study aimed to assess the outcome of locally advanced cervical and vaginal cancer treated with high-dose-rate interstitial brachytherapy (HDRB). METHODS AND MATERIALS: Between 1998 and 2004, 16 previously unirradiated patients with locally advanced cervical and vaginal cancer not suitable for intracavitary brachytherapy because of distorted anatomy or extensive vaginal disease were treated with HDRB in combination with external beam radiotherapy. All patients received whole pelvis external beam radiation therapy (EBRT) followed by interstitial implantation. The median whole pelvis external beam dose was 45 Gy (range, 39.6-50.4 Gy) with 11 patients receiving parametrial boost to a median dose of 9 Gy. Twelve (75%) of these patients received chemotherapy concurrent with external beam. All patients received a single HDRB procedure using a modified Syed-Neblett template. A CT scan was performed postimplant for needle placement verification and treatment planning purpose. Dose was prescribed to the tumor volume based on the radiographic and clinical examination. All patients received 18.75 Gy in five fractions delivered twice daily. The median followup was 25 months (6-69 months). RESULTS: Median cumulative biologic effective dose (EBRT+HDRB) to tumor volume was 78.9 Gy10 with the range of 72.5-85.2Gy10. Median cumulative biologic effective dose for the rectum and bladder were 99.4 Gy3 (range, 79.6-107.8 Gy3) and 96.4 Gy3 (range, 78.3-105.3 Gy3), respectively. Complete response was achieved in 13 (81%) patients with 3 patients having persistent disease. Five of these 13 patients developed recurrence at a median time of 14 months (distant in 4 and local and distant in 1). The 5-year actuarial local control and cause-specific survival were 75% and 64%, respectively. In subset analysis, 5-year actuarial local control was 63% for cervical cancer patients and 100% for vaginal cancer patients. No patient had acute Grade 3 or 4 morbidity. Grade 3 or 4 delayed morbidity resulting from treatment occurred in 1 patient with 5-year actuarial rate of 7%. Three patients had late Grade 2 rectal morbidity and 1 patient had Grade 2 small bowel morbidity. CONCLUSIONS: Our series suggests that single interstitial implantation procedure with five fractions of 3.75 Gy each to target volume is an effective and safe fractionation schedule. The integration of imaging modality helps in decreasing dose to the critical organs. Additional patients and followup are ongoing to determine the long-term efficacy of this approach. PMID- 17118314 TI - Comparative assessment of doses to tumor, rectum, and bladder as evaluated by orthogonal radiographs vs. computer enhanced computed tomography-based intracavitary brachytherapy in cervical cancer. AB - PURPOSE: To carry out a comparative assessment of intracavitary brachytherapy (ICBT) doses to tumor, bladder, and rectum based on orthogonal films and contrast enhanced computed tomography (CECT). METHODS AND MATERIALS: Fifty-five ICBT procedures with CT/MRI compatible applicator and CECT scans were evaluated. Doses to Point A, International Commission on Radiation Units and Measurement (ICRU) reference points for maximum bladder (B max(ICRU)) and rectum (R max(ICRU)) localized from orthogonal films were compared with CECT delineated tumor, bladder (B max(CECT)), and rectum (R max(CECT)) doses, respectively. The 95th and 90th percentile bladder (B 95(CECT) and B 90(CECT)) and rectal (R 95(CECT) and R 90(CECT)) doses based on CECT were also estimated. RESULTS: Mean percentage tumor volume encompassed within the prescribed dose of 600 cGy to Point A was 88.8%. Mean B max(ICRU), B max(CECT), R max(ICRU), and R max(CECT) were 631.3 cGy, 1221.4 cGy, 454.8 cGy, and 526.9 cGy, respectively. Paired mean differences were significant between B max(ICRU) and B max(CECT) or B 95(CECT) (both p < 0.001); R max(ICRU) and R max(CECT) (p = 0.005) or R 90(CECT) (p < 0.001), whereas insignificant for B max(ICRU) and B 90(CECT) (p = 0.281), and R max(ICRU) and R 95(CECT) (p = 0.372). CONCLUSIONS: Prescription based on Point A ICBT doses could lead to uncertainty and underdosage in tumor. ICRU 38 maximum bladder and rectal doses significantly underestimate the maximum doses to these organs and represent the 90th and 95th percentile of the maximum doses to these organs, respectively. PMID- 17118315 TI - Evaluation of the surface radiation dose and dose gradient in early stage breast cancer using high-dose-rate brachytherapy MammoSite applicator. AB - PURPOSE: To monitor the radiation dose delivered and dose homogeneity to the skin and adjacent tissues around a MammoSite applicator. METHODS AND MATERIALS: Radiation surface dose was measured on 67 consecutive patients with Stage T1 T2N0M0 breast cancer implanted with the MammoSite balloon. Thermoluminescent dosimeters and metal oxide semiconductor field effect transistors were used to measure surface dose directly over the balloon, contralateral breast, thyroid, axilla, and inframammary fold. The dose homogeneity was retrospectively studied from the treatment plans in 25 patients. RESULTS: The mean maximum skin dose was 267 cGy, with a range of 56-488 cGy per fraction, 4 cGy (+/-2 cGy) to the contralateral breast, 8 cGy (+/-4 cGy) to the thyroid, 47 cGy to the ipsilateral axilla (+/-33 cGy), and 52 cGy (+/-47 cGy) to the ipsilateral inframmammary fold. The mean dose gradients were 127%, 110%, 99%, 96%, and 89% for inflated volumes of 35, 40, 50, 60, and 70 mL, respectively. CONCLUSIONS: All patients completed therapy with no or minimal skin reaction with radiation doses comparable to or better to external beam radiation. Considering the dose homogeneity, the larger inflated volume allows for better homogeneity of dose distribution, minimizes the volume of contiguous tissue that receives relatively high doses, and may reduce the risk of fat necrosis. PMID- 17118316 TI - Lung and cardiac tissue doses in left breast cancer patients treated with single source breast brachytherapy compared to external beam tangent fields. AB - PURPOSE: We performed a retrospective analysis of doses to normal critical structures in 26 women with left-sided breast tumors treated with a high-dose rate single source brachytherapy (SSB) system, versus whole breast external beam tangent fields (EBTF) using a computer model. METHODS AND MATERIALS: Each patient's EBTF radiation plan consisted of ipsilateral tangent fields normalized to 46 Gy. A complementary SSB radiation plan was designed to deliver 34 Gy in 10 fractions at 1cm into tissue enclosing a sphere of diameter 4.6 cm at tumor bed, simulating our institution's experience with the MammoSite applicator. Critical structure volumes and doses analyzed included the volume of whole heart and left ventricle receiving >25 Gy, and left lung (LL) receiving 20 and 13 Gy for EBTF plans. For SSB we determined the linear quadratic 2-Gy-equivalent high-dose-rate doses of 23.4, 20, and 14 Gy to be equivalent to 25, 20, and 13 Gy, respectively. We then compared the two doses for each critical structure. RESULTS: The percent of whole heart receiving >25 Gy equivalent using EBTF and SSB was 2% and 0% (mean dose 2.52 vs. 1.65 Gy), left ventricle receiving >25 Gy using EBTF and SSB was 3% and 0% (mean dose 3.78 vs. 2.68 Gy), LL receiving >20 Gy using EBTF and SSB was 11.5% and 0.5%, and LL receiving >13 Gy using EBTF and SSB was 12.6% and 1.85% (mean dose to LL was 6.06 Gy for EBTF vs. 2.25 Gy for SSB), respectively. CONCLUSIONS: When compared to EBTF, breast brachytherapy with SSB is associated with a significant decrease in mean dose and percentage volume receiving a given dose for each organ. PMID- 17118317 TI - A descriptive analysis of postimplant dosimetric parameters from Radiation Therapy Oncology Group P0019. AB - BACKGROUND: To date, there are few descriptive analyses of postimplant dosimetry from multi-institutional clinical trials. The purpose of this report is to describe the postimplant dosimetry achieved in Radiation Therapy Oncology Group (RTOG) 0019. METHODS AND MATERIALS: Patients were treated with external beam radiation therapy (45 Gy/25 fractions) followed by a prostate implant (I-125, prescription dose 108 Gy). Postimplant dosimetric assessment was accomplished by obtaining a CT scan of the prostate 1 month after the date of the implant procedure. Prostate volume was outlined by the first author. Dose-volume histograms were calculated by the Radiologic Physics Center. Four dosimetric quantifiers (DQs) were examined: D90 is the dose (reported as percentage of the prescription dose) received by 90% of the prostate; V100, V150, V200 is the percentage of the prostate volume receiving 100%, 150%, and 200% of the prescription dose, respectively. For the purposes of analysis, institutions were divided into three groups according to accrual (<5, 6-9, 10-12). RESULTS: One hundred thirty-eight patients from 27 institutions were registered in the study. Nineteen patients were excluded from this analysis; 14 who had no data and 5 who were ineligible, leaving 119 for analysis. The mean, median, and range of the four DQs are as follows: D90 105.6%, 106.0%, 57.6-174.8%; V100 89.8%, 92.6%, 11.2 100%; V150 58.4%, 59.6%, 0.9-93.7%; and V200 27.9%, 25.1%, 0.3-85.2%. Statistically significant differences according to institutional accrual were observed for D90 (p = 0.0283) and V200 (p = 0.0075), but not for V100 (p = 0.1534) and V150 (p = 0.0509). CONCLUSIONS: The DQ observed in this multi institutional prospective study are roughly comparable to series from single institutions with considerable brachytherapy experience. Differences in DQs were observed according to institutional accrual. These data could be used to determine a community standard with respect to postimplant dosimetry. PMID- 17118318 TI - A dosimetric analysis of unstranded seeds versus customized stranded seeds in transperineal interstitial permanent prostate seed brachytherapy. AB - PURPOSE: This study aimed to retrospectively analyze the dosimetric and toxicity results from 272 patients with localized prostate cancer treated consecutively using loose or stranded radioactive seeds by transrectal ultrasound-guided transperineal permanent prostate seed brachytherapy. METHODS AND MATERIALS: Two hundred seventy-two patients with localized prostate cancer treated between February 2002 and June 2004 were analyzed. All patients were treated with radioactive iodine-125 or palladium-103 using unstranded or loose seeds (USS) (159 patients) or customized stranded seeds (CSS) at variable spacing (5-50 mm) (113 patients) (Vari-Strand; BrachySciences, Oxford, CT). A single experienced brachytherapist performed all implants. RESULTS: There was a slight improvement in the dosimetric parameter D90 between the CSS (101.9%) and USS (99.3%) groups (p = 0.041). However, overall implant quality based on Radiation Therapy Oncology Group (RTOG) guidelines was similar between both groups. CONCLUSIONS: We conclude that the D90 value calculated for CSS is statistically improved when compared to the USS cohort, but without a clinically significant difference. There was no difference in the toxicity scores in either group. Overall quality between groups is comparable in our institution. PMID- 17118319 TI - Comparative study of dosimetry between high-dose-rate and permanent prostate implant brachytherapies in patients with prostate adenocarcinoma. AB - PURPOSE: To compare the dose coverage, conformity, and homogeneity between high dose-rate (HDR) brachytherapy and permanent prostate implant (PPI) in the treatment of prostate adenocarcinoma. METHODS AND MATERIALS: From January 2003 to August 2004, 54 patients (108 implants) underwent HDR brachytherapy of prostate cancer with iridium-192 stepping source. Of patients who underwent PPI brachytherapy with iodine-125, 72 patients were randomly selected for the purpose of dosimetric comparison. PPI preplan was done based on transrectal ultrasound study, and postplan was done using CT 1 month after implant. Dosimetric parameters of HDR were compared to that of PPI preplan and postplan. RESULTS: HDR brachytherapy had lower D90 (111.5% vs. 120.2%), lower V100 (97.2% vs. 99.6%), lower natural dose ratio (1.03 vs. 1.13), higher conformal index (0.69 vs. 0.62), and higher homogeneity index (0.63 vs. 0.52) than PPI preplan (all p < 0.0001). All the dosimetric parameters of PPI postplan including D90 (86.7%), V100 (82.0%), natural dose ratio (0.92), conformal index (0.53), and homogeneity index (0.42) were inferior to HDR brachytherapy (all p < 0.0001). CONCLUSIONS: HDR brachytherapy of the prostate can provide better dose coverage, conformity, and homogeneity compared to PPI. PMID- 17118320 TI - A prospective dose escalation trial of high-dose-rate brachytherapy boost for prostate cancer: Evidence of hypofractionation efficacy? AB - PURPOSE: The study aimed to evaluate mature outcomes of a Phase I/II high-dose rate brachytherapy (HDRB) boost protocol. METHODS AND MATERIALS: We analyzed data from 88 patients with T1a-T3a, N0, M0 prostate adenocarcinoma treated on a prospective Phase I/II HDRB boost protocol of 16 (n = 47) or 20 Gy (n = 41) in four fractions, without planned androgen deprivation therapy. HDRB was added to 46 Gy of external beam radiotherapy (EBRT). Outcomes were compared to a contemporaneous retrospective cohort of 104 patients receiving 66 Gy EBRT monotherapy. The primary endpoint was freedom from biochemical failure, defined as a 2 ng/mL rise above the lowest prostate-specific antigen (PSA) (FFbFn2), whereas the American Society of Therapeutic Radiology and Oncology consensus definition (ACD) was used for comparative purposes. RESULTS: For the HDRB cohort, the overall actuarial 5-year FFbFn2 was 67.4% (95% CI: 58.2-75.5%). For the HDRB doses of 16 and 20 Gy, the 5-year FFbFn2 rates were 58.8% (95% CI: 41.9-72.5%) and 77.3% (95% CI: 64.4-88.3%), respectively (log-rank test p = 0.07). Compared to men treated with 66 Gy EBRT, using multivariate analysis, there was no significant benefit to using HDRB with the FFbFn2 outcome (p = 0.52), yet the ACD suggested a significant advantage (hazard ratio 0.50, 95% CI: 0.29-0.86, p = 0.011). There was a trend to better FFbFn2 outcomes with increasing biologically effective doses (p = 0.09), which was significant using the ACD (p = 0.0003). CONCLUSIONS: The data support HDRB boost as a potential means of dose escalation in prostate cancer. Significant findings using the ACD need to be validated with contemporary biochemical failure definitions. Prospective trials to optimize fractionation and evaluate outcomes in comparison to contemporary EBRT techniques are warranted. PMID- 17118321 TI - Prostate brachytherapy seed migration to the right coronary artery associated with an acute myocardial infarction. AB - PURPOSE: We report a case of prostate brachytherapy seed migration to the right coronary artery (RCA) associated with an acute myocardial infarction (AMI). METHODS AND MATERIALS: A 69-year-old male was diagnosed with Prostate Adenocarcinoma Stage II (T(1c)N0M0) in October 2003. He underwent percutaneous transperineal interstitial permanent prostate brachytherapy with the implantation of 94 loose iodine (125I) seeds under transrectal ultrasound guidance on 15 December, 2003. The implantation was designed to deliver a minimum dose of 144 Gy. RESULTS: Postimplant pelvic radiography at Day 30 showed five seeds missing. No chest radiography was done until hospital admission on October 10, 2005 for AMI. Cine radiography from cardiac catheterization revealed 86 metallic seeds remaining in the pelvic region, 4 that had migrated to the lungs (2 left and 2 right) and 2 to the heart. Two seeds were unaccounted for. Of the two cardiac seeds, one was lodged in the right ventricle endocardium and the other in the midsegment of the RCA at the site of a severely stenotic lesion that resulted in an AMI. CONCLUSIONS: To our knowledge, this is the first case of brachytherapy seed migration to the RCA associated with an AMI. Echocardiography before brachytherapy seed implantation might be considered in patients felt to be at high risk for cardiac shunting. PMID- 17118322 TI - Difficulties with quasi-induced exposure when speed varies systematically by vehicle type. AB - The attractiveness of quasi-induced exposure lies in the simplistic nature of its theory and application. As opposed to vehicle miles of travel, quasi-induced exposure is developed solely from the accident data themselves. Involvement ratios (IRs) are used to describe the relative over- or under-involvement of different driver-vehicle combinations in traffic accidents. The issue of systematic bias in the involvement ratios is explored, and it is shown both theoretically and empirically that the variation in average speeds between vehicle types can affect relative accident involvement ratios. For vehicle types that routinely travel faster, the IRs will likely be underestimated; while for slower-moving vehicle types, the IRs will be overestimated. In the case of speed, the magnitude of the effect on the IRs is dependent both on the magnitude of the speed differential and the percentage of slower vehicles in the traffic stream. Conclusions can be extended to whenever there are speed or other behavior disparities associated with the driver-vehicle combinations in the traffic stream. Other examples include the speed discrepancy caused by different drivers (e.g., younger versus older drivers). Caution must be used in interpreting results from applications of the quasi-induced technique whenever such biases might be encountered. PMID- 17118323 TI - Safety effects of roundabouts in Flanders: signal type, speed limits and vulnerable road users. AB - This paper examines the road safety effects of roundabouts built in Flanders between 1994 and 2000. While the overall effect is positive (39% reduction of injury accidents), the results vary considerably with the speed limit on the main and adjacent road (the higher, the more effective) and the pre-roundabout signalization of the intersection (32% reduction with traffic lights versus 44% without). However, microscopic analysis reveals that roundabouts are not always effective. Serious injury accidents are estimated to increase by 117% on 70 km/h x 50 km/h intersections equipped with signalization before the roundabout. The number of injury accidents involving vulnerable road users is also found to increase (28%) on 50 km/h x 50 km/h junctions that were originally signalized. Moreover, the vulnerable road user is more likely to get fatally or seriously injured. Therefore, it is concluded that traffic lights protect vulnerable road users more effectively than roundabouts, which, in turn, are superior to intersections without signalization. PMID- 17118324 TI - Molecular characterization and species identification of the Anopheles dirus and An. minimus complexes in north-east India using r-DNA ITS-2. AB - The sibling species composition of the Anopheles minimus and Anopheles dirus complexes is poorly known in the highly malarious north-eastern region of India where these two vector taxa are accountable for most of the malaria transmission among 30.7 million inhabitants. Prevalent members of these two complexes in this part of India were identified using sequences for the second internal transcribed spacer (ITS2) of ribosomal DNA. Anopheles baimaii (species D) of the An. dirus complex and An. minimus s.s. (species A) of the An. minimus complex were detected in Arunachal Pradesh, Assam, Meghalaya and Nagaland states. No intraspecific variation was observed in the ITS2 sequence (479bp) of An. baimaii whereas a single substitution was detected in the ITS2 sequence (372bp) of An. minimus from Nagaland state. PMID- 17118325 TI - History of Tropical Medicine William Ernest Cooke, FRCSI (1879-1967) and his Asian tour of 1929-1930. AB - Dr. W.E. Cooke was not one of the most distinguished of tropical physicians, but the specialty was heavily dependent on his likes. After several junior appointments, he was Superintendent of the Hospital for Tropical Diseases in London. He carried out an extensive tour of Asia in 1929-1930 and kept careful daily notes throughout this period, giving a valuable insight into the challenges faced by the pioneering tropical physicians and surgeons from European countries. PMID- 17118326 TI - Ultraviolet fluorescence photography: patterns in established pterygia. AB - PURPOSE: To investigate established pterygia using our newly developed ultraviolet fluorescence photography (UVFP) system. DESIGN: Prospective observational case series. METHODS: setting: Prince of Wales Hospital, Sydney, Australia. study population: Fourteen patients (both eyes) attending the Ophthalmology Clinic at Prince of Wales Hospital for assessment of their established pterygia. There were eight men and six women, with an age range of 26 to 62 years. A total of 15 (75%) of 20 had primary pterygia, and five (25%) of 20 had recurrent pterygia. There were no specific exclusion criteria. observation procedures: Ultraviolet and standard (control) photographs were taken of the nasal and temporal interpalpebral regions bilaterally. main outcome measures: The presence of established pterygia detected by standard photography and the corresponding presence and patterns of areas of fluorescence detected by UVFP. RESULTS: In the 14 patients, 20 established pterygia were identified on standard photography. On UVFP, four patterns of fluorescence of established pterygia were identified. Of the 20 pterygia, six (30%) of 20 demonstrated fluorescence at the leading edge of the pterygium, seven (35%) of 20 demonstrated fluorescence at the limbus, three (15%) of 20 demonstrated fluorescence at both the leading edge and the limbus, and four (20%) of 20 demonstrated no visible fluorescence. CONCLUSIONS: In this study, we describe patterns of fluorescence in established pterygia by UVFP. We hypothesize that the areas of fluorescence represent areas of cellular activity within the pterygium. The patterns of fluorescence may be useful to further understand of pterygium growth and pathogenesis. PMID- 17118327 TI - A substrate for deubiquitinating enzymes based on time-resolved fluorescence resonance energy transfer between terbium and yellow fluorescent protein. AB - Deubiquitinating enzymes (DUBs) proteolytically cleave ubiquitin from ubiquitinated proteins, and inhibition of DUBs that rescue oncogenic proteins from proteasomal degradation is of emerging therapeutic interest. Recently, USP2 and UCH37 have been shown to deubiquitinate tumor-growth-promoting proteins, and other DUBs have been shown to be overexpressed in cancer cells. Therefore inhibition of DUBs is of interest as a potential therapeutic strategy for treating cancer. DUBs require the presence of properly folded ubiquitin protein in the substrate for efficient proteolysis, which precludes the use of synthetic peptide substrates in DUB activity assays. Because of the requirement for full length ubiquitin, substrates suitable for use in fluorescent assays to identify or study DUB inhibitors have been difficult to prepare. We describe the development of a time-resolved fluorescence resonance energy transfer (FRET) based DUB substrate that incorporates full-length ubiquitin that is site specifically labeled using genetically encoded yellow fluorescent protein (YFP) and a chemically attached terbium donor. The intact substrate shows a high degree of FRET between terbium and YFP, whereas DUB-dependent cleavage leads to a decrease in FRET. PMID- 17118328 TI - Evidence for a large conductance voltage gated cationic channel in rough endoplasmic reticulum of rat hepatocytes. AB - In this work, we report the single channel characterization of a voltage gated cationic channel from rough endoplasmic reticulum (RER) membranes of rat hepatocytes incorporated into a planar lipid bilayer. The channel was found to be cation selective with a main conductance of 598+/-20 pS in 200 mM KCl cis/50 mM KCl trans. The channel open probability appeared voltage dependent with a voltage for half activation (V(1/2)) of 38 mV and an effective gating charge z of -6.66. Adding either 4-AP (5 mM) or ATP (2.5 mM) to the side corresponding to the cell internal medium caused a strong inhibition of the channel activity. This channel is likely to be involved in maintaining proper cation homeostasis in the RER of hepatocytes. PMID- 17118329 TI - Cinnamtannin B-1 from bay wood reduces abnormal intracellular Ca2+ homeostasis and platelet hyperaggregability in type 2 diabetes mellitus patients. AB - Type 2 diabetes mellitus induces a number of cardiovascular disorders, including platelet hyperactivity and hyperaggregability, which is associated to an increased oxidant production and abnormal cytosolic Ca2+ mobilization. In the present study, we have investigated the effect of cinnamtannin B-1 obtained from bay wood on oxidants production, Ca2+ mobilization and aggregation in platelets from type 2 diabetic donors. Pretreatment of platelets with cinnamtannin B-1 reversed the enhanced oxidants production and Ca2+ mobilization, including Ca2+ entry, evoked by thapsigargin plus ionomycin or thrombin, observed in platelets from diabetic subjects, so that in the presence of cinnamtannin B-1 Ca2+ entry was similar in platelets from healthy and diabetic subjects. In addition, cinnamtannin B-1 reduced thrombin-induced aggregation in platelets from type 2 diabetic subjects. We conclude that cinnamtannin B-1 exerts an effective antioxidant action in platelets from patients with type 2 diabetes mellitus and reverses the enhanced Ca2+ mobilization and hyperaggregability. PMID- 17118330 TI - Role of intracellular calcium and phospholipase A2 in arachidonic acid-induced toxicity in liver cells overexpressing CYP2E1. AB - Liver cells (HepG2 and primary hepatocytes) overexpressing CYP2E1 and exposed to arachidonic acid (AA) were previously shown to lose viability together with enhanced lipid peroxidation. These events were blocked in cells pre-incubated with antioxidants (alpha-tocopherol, glutathione ethyl ester), or in HepG2 cells not expressing CYP2E1. The goal of the current study was to evaluate the role of calcium and calcium-activated hydrolases in these CYP2E1-AA interactions. CYP2E1 expressing HepG2 cells treated with AA showed an early increase in cytosolic calcium and partial depletion of ionomycin-sensitive calcium stores. These changes in calcium were blocked by alpha-tocopherol. AA activated phospholipase A2 (PLA2) in CYP2E1-expressing liver cells, and this was inhibited by PLA2 inhibitors or alpha-tocopherol. PLA2 inhibitors prevented the cell death caused by AA, without affecting CYP2E1 activity or lipid peroxidation. AA toxicity and PLA2 activation were inhibited in calcium-depleted cells, but not by removal of extracellular calcium alone. Removal of extracellular calcium inhibited the early increase in cytosolic calcium caused by AA. CYP2E1 overexpressing HepG2 cells exposed to AA showed a decrease in mitochondrial membrane potential, which was prevented by the PLA2 inhibitors. These results suggest that AA-induced toxicity to CYPE1-expressing cells: (i) is associated with release of Ca2+ from intracellular stores that depends mainly on oxidative membrane damage; (ii) is associated with activation of PLA2 that depends on intracellular calcium and lipid peroxidation; (iii) does not depend on increased influx of extracellular calcium, and (iv) depends on the effect of converging events (lipid peroxidation, intracellular calcium, activation of PLA2) on mitochondria to induce bioenergetic failure and necrosis. These interactions may play a role in alcohol liver toxicity, which requires polyunsaturated fatty acids, and involves induction of CYP2E1. PMID- 17118331 TI - Glycogen metabolism in tissues from a mouse model of Lafora disease. AB - Laforin, encoded by the EPM2A gene, by sequence is a member of the dual specificity protein phosphatase family. Mutations in the EPM2A gene account for around half of the cases of Lafora disease, an autosomal recessive neurodegenerative disorder, characterized by progressive myoclonus epilepsy. The hallmark of the disease is the presence of Lafora bodies, which contain polyglucosan, a poorly branched form of glycogen, in neurons, muscle and other tissues. Glycogen metabolizing enzymes were analyzed in a transgenic mouse over expressing a dominant negative form of laforin that accumulates Lafora bodies in several tissues. Skeletal muscle glycogen was increased 2-fold as was the total glycogen synthase protein. However, the -/+glucose-6-P activity of glycogen synthase was decreased from 0.29 to 0.16. Branching enzyme activity was increased by 30%. Glycogen phosphorylase activity was unchanged. In whole brain, no differences in glycogen synthase or branching enzyme activities were found. Although there were significant differences in enzyme activities in muscle, the results do not support the hypothesis that Lafora body formation is caused by a major change in the balance between glycogen elongation and branching activities. PMID- 17118332 TI - Mapping von Willebrand factor A domain binding sites on a snake venom metalloproteinase cysteine-rich domain. AB - The PIII class of the snake venom metalloproteinases (SVMPS) are acknowledged to be one of the major hemorrhage producing toxins in crotalid venoms. This class of SVMPS are structurally distinguished by the presence of disintegrin-like and cysteine-rich domains carboxy to the metalloproteinase domain and thus share structural homology with many of the ADAMs proteins. It has been suggested that the presence of the carboxy domain are the key structural determinants for potent hemorrhagic activity in that they may serve to target the proteinases to specific key extracellular matrix and cell surface substrates for proteolysis leading to hemorrhage production at the capillaries. Following from previous studies in our laboratory in this investigation we scanned the cysteine-rich domain of the PIII hemorrhagic SVMP jararhagin using synthetic peptides in an attempt to identify regions which could bind to von Willebrand factor (vWF), a known binding partner for jararhagin. From these studies we identified two such peptide, Jar6 and Jar7 that could support binding to vWF as well as block the recombinant cysteine-rich domain of jararhagin binding to vWF. Using the coordinates for the recently solved crystal structure of the PIII SVMP VAP1, we modeled the structure of jararhagin and attempted to dock the modeled cysteine-rich structure of that protein to the A1 domain of vWF. These studies indicated that effective protein protein interaction between the two ligands was possible and supported the data indicating that the Jar6 peptide was involved, whereas the Jar7 peptide was observed to be sterically blocked from interaction. In summary, our studies have identified a region on the cysteine-rich domain of a PIII SVMP that interacts with vWF and based on molecular modeling could be involving in the interaction of the cysteine-rich domain of the SVMP with the A1 domain of vWF thus serving to target the toxin to the protein for subsequent proteolytic degradation. PMID- 17118333 TI - Purification and characterization of a lectin from endophytic fungus Fusarium solani having complex sugar specificity. AB - A lectin from the mycelial extract of an endophytic strain of Fusarium solani was purified. Its hemagglutinating activity was inhibited by glycoproteins possessing N-linked as well as O-linked glycans. The thermodynamics and kinetics of binding of glycans and glycoproteins to F. solani lectin was studied using surface plasmon resonance. The lectin showed high affinity for asialofetuin, asialomucin, asialofibrinogen, and thyroglobulin; and comparatively low affinity for mucin, fetuin, fibrinogen, and holotransferrin. Glycoproteins showed several fold higher affinity than their corresponding glycans with significant contribution from enthalpy and positive entropy, suggesting the involvement of non-polar protein protein interaction. Moreover, the higher affinity of the glycoproteins was due to their faster association rates and low activation energy. PMID- 17118334 TI - Effects of autonomic agonists and immunomodulatory cytokines on polymeric immunoglobulin receptor expression by cultured rat and human salivary and colonic cell lines. AB - OBJECTIVE: Immunoglobulin A (IgA) is transported across glandular epithelial cells by polymeric immunoglobulin receptor (plgR), with each receptor molecule participating in only one round of transcytosis. Nerve-related stimuli rapidly increase salivary secretion of IgA, while concentrations are increased in the autoimmune disease Sjogren's syndrome. Our aim here was to determine whether autonomic agonists and cytokines present in Sjogren's-affected glands can up regulate salivary cell plgR expression. METHODS: Cultures of rat parotid acinar cells (PAR C5) and human submandibular gland ductal cells (HSG) were exposed to carbachol or adrenaline for 24 h and to interleukin-4 and/or interferon-gamma for 48 h. The human colonic cell line HT-29 served as a positive control for cytokine response. plgR mRNA was quantified by reverse transcription and real-time PCR and protein expression was examined by immunoblotting. RESULTS: Carbachol increased plgR mRNA levels significantly in all cells but adrenaline did so only with PAR cells (P<0.05). HSG and HT-29 cells both up-regulated plgR gene transcription on exposure to interleukin-4 and interferon-gamma either alone or in combination (P<0.05). By contrast, production of plgR mRNA in PAR cells tended to decrease in response to all cytokine treatments. plgR protein levels rose in line with mRNA expression in cytokine-treated HT-29 cultures (P<0.05). CONCLUSIONS: Autonomimetics can up-regulate plgR transcription in transformed and neoplastic salivary and colonic cells, although intracellular coupling mechanisms require further investigation. Immunomodulatory cytokines increased plgR expression in one of the salivary cell lines, but additional work is needed to establish whether this occurs in Sjogren's patients. PMID- 17118335 TI - Abnormal dentin structure in two novel gene mutations [COL1A1, Arg134Cys] and [ADAMTS2, Trp795-to-ter] causing rare type I collagen disorders. AB - Histological and ultrastructural observations of dentin of two patients affected with rare types of type I collagen disorders are presented. In the first case, a homozygous nonsense mutation in ADAMTS2 (substitution of a codon for tryptophan by a stopcodon) causes type VIIC Ehlers-Danlos syndrome (EDS) with multiple tooth agenesis and focal dysplastic dentin defects. In the second case, a missense mutation in COL1A1 (substitution of arginine by cysteine) results in a type I EDS phenotype with clinically normal-appearing dentition. Tooth samples are investigated by using light microscopy (LM), transmission electron microscopy (TEM) and immunostaining for types I and III collagen, and tenascin. These are compared with samples from patients with types III and IV osteogenesis imperfecta (OI) in association with dentinogenesis imperfecta (DI), showing a consistently abnormal appearance of the dentin in all specimens, with variations being primarily those of degree of change. Similarities in histological changes include the alternating presence of normal and severe pathological areas in primary and secondary dentin, the latter being characterized by large canal-like structures in atubular areas. Ultrastructural evidence of pathological dentinogenesis include abnormal distribution, size and organization of collagen fibers, which may also be found in clinically unaffected teeth. The histological and ultrastructural changes seen can be explained on the basis of odontoblast dysfunction which may be secondary to the collagen defect, interfering with different levels of odontoblast cell function and intercellular communication. These observations on (ultra)structural dentin defects associated with the two novel gene mutations are the first ever reported. PMID- 17118336 TI - Anti-tumor effects of dehydroaltenusin, a specific inhibitor of mammalian DNA polymerase alpha. AB - In the screening of selective inhibitors of eukaryotic DNA polymerases (pols), dehydroaltenusin was found to be an inhibitor of pol alpha from a fungus (Alternaria tennuis). We succeeded in chemically synthesizing dehydroaltenusin, and the compound inhibited only mammalian pol alpha with IC50 value of 0.5 microM, and did not influence the activities of other replicative pols such as pols delta and epsilon, but also showed no effect on pol alpha activity from another vertebrate, fish, or from a plant species. Dehydroaltenusin also had no influence on the other pols and DNA metabolic enzymes tested. The compound also inhibited the proliferation of human cancer cells with LD50 values of 38.0-44.4 microM. In an in vivo anti-tumor assay on nude mice bearing solid tumors of HeLa cells, dehydroaltenusin was shown to be a promising suppressor of solid tumors. Histopathological examination revealed that increased tumor necrosis and decreased mitotic index were apparently detected by the compound in vivo. Therefore, dehydroaltenusin could be of interest as not only a mammalian pol alpha-specific inhibitor, but also as a candidate drug for anti-cancer treatment. PMID- 17118338 TI - A protective role for the polyamine spermine against drought stress in Arabidopsis. AB - Cellular polyamine content often changes in response to abiotic stresses. However, its physiological relevance is unknown. We found that an Arabidopsis mutant plant (acl5/spms), which cannot produce spermine, is hypersensitive to high salt. Examination of drought sensitivity of the mutant and comparison with wild type plants indicated hypersensitivity to drought. This phenotype was cured by spermine pretreatment but not by the other polyamines putrescine and spermidine, suggesting that drought-hypersensitivity exhibited by the mutant is due to spermine deficiency. The water loss rate of wild type and mutant plants were similar until 20 min after onset of dehydration stress, but after a longer exposure the rate in mutant plants was higher than in wild type plants. Consistent with this result, the stomata of the mutant leaves remained open while in wild type leaves they closed. Based on the collected data, we discuss a role for spermine in response to drought stress. PMID- 17118337 TI - Peptide modulators of Src activity in G1 regulate entry into S phase and proliferation of NIH 3T3 cells. AB - Cascades of kinases and phosphatases are regulated by selective protein-protein interactions that are essential for signal transduction. Peptide modulators of these interactions have been used to dissect the function of individual components of the signaling cascade, without relying on either the over- or underexpression of proteins. Previously, we identified RACK1 as an endogenous substrate, binding partner and inhibitor of Src tyrosine kinases. Here, we utilized cell-permeable peptides that selectively disrupt or enhance the interaction of RACK1 and Src to further examine the function of RACK1. Our results provide direct physiologic evidence that RACK1 regulates growth of NIH3T3 cells by suppressing the activity of Src and other cell cycle regulators in G1, and delaying entry into S phase. They also demonstrate the potential for using peptide modulators of Src activity as a tool for regulating cell growth, and for designing new strategies for cancer therapy that target specific protein-protein interactions. PMID- 17118339 TI - Characterization of the cardiac sodium channel SCN5A mutation, N1325S, in single murine ventricular myocytes. AB - The N(1325)S mutation in the cardiac sodium channel gene SCN5A causes the type-3 long-QT syndrome but the arrhythmogenic trigger associated with N(1325)S has not been characterized. In this study, we investigated the triggers for cardiac events in the expanded N(1325)S family. Among 11 symptomatic patients with document triggers, six died suddenly during sleep or while sitting (bradycardia induced trigger), three died suddenly, and two developed syncope due to stress and excitement (non-bradycardia-induced). Patch-clamping studies revealed that the late sodium current (I(Na,L)) generated by mutation N(1325)S in ventricular myocytes from TG-NS/LQT3 mice was reduced with increased pacing, which explains bradycardia-induced mortalities in the family. The non-bradycardic triggers are related to the finding that APD became prolonged and unstable at increasing rates, often with alternating repolarization phases which was corrected with verapamil. This implies that Ca2+ influx and intracellular Ca2+ ([Ca2+]i) ions are involved and that [Ca2+]i inhomogeneity may be the underlying mechanisms behind non-bradycardia LQT3 arrhythmogenesis associated with mutation N(1325)S. PMID- 17118340 TI - Palmitate activates AMP-activated protein kinase and regulates insulin secretion from beta cells. AB - AMP-activated protein kinase (AMPK) is an energy sensor that regulates cellular metabolism. Changes in AMPK activity contribute to the regulation of insulin secretion. Epidemiological evidence links the ingestion of saturated fatty acid with hyperinsulinemia. The aim of the present study was to examine the effects of palmitate on beta cell AMPK activity and insulin secretion. Isolated rat islets and MIN6 beta cells were treated acutely (5-60 min) or chronically (24 h) with palmitate. Insulin secretion, AMPK and acetyl CoA carboxylase phosphorylation were assessed. The acute effects of palmitate included AMPK activation and augmentation in insulin secretion. Activation of AMPK by 24h pretreatment with palmitate suppressed glucose-stimulated insulin secretion, but not the response of insulin secretion to combined stimuli of glucose and palmitate. This study demonstrated that palmitate availability affected beta cell AMPK activity. In beta cells, an increase in AMPK activity may be required for fatty acid-induced fatty acid oxidation and prevention of lipotoxicity. PMID- 17118341 TI - CD133+ hepatic stellate cells are progenitor cells. AB - Hepatic stellate cells (HSC) play an important role in the development of liver fibrosis. Here, we report that HSC express the stem/progenitor cell marker CD133 and exhibit properties of progenitor cells. CD133+ HSC of rats were selected by specific antibodies and magnetic cell sorting. Selected cells displayed typical markers of HSC, endothelial progenitor cells (EPC), and monocytes. In cell culture, CD133+ HSC transformed into alpha-smooth muscle actin positive myofibroblast-like cells, whereas application of cytokines known to facilitate EPC differentiation into endothelial cells led to the formation of branched tube like structures and induced expression of the endothelial cell markers endothelial nitric oxide synthase and vascular-endothelial cadherin. Moreover, cytokines that guide stem cells to develop hepatocytes led to the appearance of rotund cells and expression of the hepatocyte markers alpha-fetoprotein and albumin. It is concluded that CD133+ HSC are a not yet recognized progenitor cell compartment with characteristics of early EPC. Their potential to differentiate into endothelial or hepatocyte lineages suggests important functions of CD133+ HSC during liver regeneration. PMID- 17118342 TI - Chain length dependence of the interactions of bisquaternary ligands with the Torpedo nicotinic acetylcholine receptor. AB - The interactions of a series of bisholine esters [(CH3)3N+CH(2)CH2OCO-(CH2)n COOCH2CH2N+(CH3)3] with the Torpedo nicotinic acetylcholine receptor have been investigated. In equilibrium binding studies, [3H]-suberyldicholine (n=6) binds to an equivalent number of sites as [3H]-acetylcholine and with similar affinity (KD approximately 15 nM). In competition studies, all bischoline esters examined displaced both radioligands in an apparently simple competitive manner. Estimated dissociation constants (KI) showed clear chain length dependence. Short chain molecules (n6) had high affinity similar to suberyldicholine. Functional responses were measured by either rapid flux techniques using Torpedo membrane vesicles or voltage-clamp analyses of recombinant receptors expressed in Xenopus oocytes. Both approaches revealed that suberyldicholine (EC50 approximately 3.4 microM) is 14-25-fold more potent than acetylcholine. However, suberyldicholine elicited only about 45% of the maximum response of the natural ligand, i.e., it is a partial agonist. The potency of this bischoline series increased with chain length. Whereas the shorter ligands (nor=4) had similar (or higher) potency to suberyldicholine. Ligand efficacy had an approximately bell-shaped dependence on chain length and compounds where nor=8 were very poor partial agonists. Based on estimates of interonium distances, we suggest that bisquaternary ligands can interact with multiple binding sites on the nAChR and, depending on the conformational state of the receptor, these sites are 15-20A apart. PMID- 17118343 TI - The effect of opiates on the activity of human placental aromatase/CYP19. AB - Aromatase, cytochrome P450 19, is a key enzyme in the biosynthesis of estrogens by the human placenta. It is also the major placental enzyme that metabolizes the opiates L-acetylmethadol (LAAM), methadone, and buprenorphine (BUP). Methadone and BUP are used in treatment of the opiate addict and are competitive inhibitors of testosterone conversion to estradiol (E(2)) and 16alpha-hydroxytestosterone (16-OHT) to estriol (E(3)) by aromatase. The aim of this investigation is to determine the effect of 20 opiates, which can be administered to pregnant patients for therapeutic indications or abused, on E(2) and E(3) formation by placental aromatase. Data obtained indicated that the opiates increased, inhibited, or had no effect on aromatase activity. Their effect on E(3) formation was more pronounced than that on E(2) due to the lower affinity of 16-OHT than testosterone to aromatase. The K(i) values for the opiates that inhibited E(3) formation were sufentanil, 7 +/- 1 microM; LAAM, 13 +/- 8 microM; fentanyl, 25 +/ 5 microM; oxycodone, 92 +/- 22 microM; codeine, 218 +/- 69 microM; (+) pentazocine, 225 +/- 73 microM. The agonists morphine, heroin, hydromorphone, oxymorphone, hydrocodone, propoxyphene, meperidine, levorphanol, dextrorphan, and (-)-pentazocine and the antagonists naloxone and naltrexone caused an increase in E(3) formation by 124-160% of control but had no effect on E(2) formation. Moreover, oxycodone and codeine did not inhibit E(2) formation and the IC(50) values for fentanyl, sufentanil, and (+)-pentazocine were >1000 microM. It is unlikely that the acute administration of the opiates that inhibit estrogen formation would affect maternal and/or neonatal outcome. However, the effects of abusing any of them during the entire pregnancy are unclear at this time. PMID- 17118344 TI - Irofulven induces replication-dependent CHK2 activation related to p53 status. AB - CHK2 and p53 are frequently mutated in human cancers. CHK2 is known to phosphorylate and stabilize p53. CHK2 has also been implicated in DNA repair and apoptosis induction. However, whether p53 affects CHK2 activation and whether CHK2 activation modulates chemosensitivity are unclear. In this study, we found that in response to the DNA damage agent, irofulven, CHK2 activation, rather than its expression, is inversely correlated to p53 status. Irofulven inhibits DNA replication and induces chromosome aberrations (breaks and radials) and p53 dependent cell cycle arrest. Pretreatment of cells with the DNA polymerase inhibitor, aphidicolin, resulted in reduction of irofulven-induced CHK2 activation and foci formation, indicating that CHK2 activation by irofulven is replication-dependent. Furthermore, by using ovarian cancer cell lines expressing dominant-negative CHK2 and CHK2-knockout HCT116 cells, we found that CHK2 activation contributes to the control of S and G2/M cell cycle arrests, but not chemosensitivity to irofulven. Overall, this study demonstrates that in response to irofulven-induced DNA damage, the activation of CHK2 is dependent on DNA replication and related to p53 status. By controlling cell cycle arrest and DNA replication, p53 affects CHK2 activation. CHK2 activation contributes to cell cycle arrest, but not chemosensitivity. PMID- 17118346 TI - Calmodulin potentiates G beta gamma activation of phospholipase C-beta3. AB - Phospholipase C-beta (PLC-beta) isozymes (EC 3.1.4.11) hydrolyze the membrane phospholipid phosphatidylinositol-4,5-bisphosphate to generate intracellular second messenger signaling molecules inositol-1,4,5-trisphosphate (IP3) and diacylglycerol (DAG) in response to receptor activation and other cellular stimuli. PLCbeta1 and PLCbeta3 isozymes were previously demonstrated to bind the calcium-sensitive molecule calmodulin [McCullar JS, Larsen SA, Millimaki RA, Filtz TM. Calmodulin is a phospholipase C-{beta} interacting protein. J Biol Chem 2003;278(36):33708-13]. We have now shown through fluorescence anisotropy that calmodulin/PLCbeta3 affinities increase with increasing calcium in a physiologically relevant concentration range. The bimolecular affinity constants for calmodulin interaction with PLCbeta1 or PLCbeta3 were estimated as 260 and 200 nM, respectively, from fluorescence anisotropy data. There was no effect of calmodulin on basal or G alpha q-stimulated catalytic activity for either isozyme. However, the interaction between calmodulin and PLCbeta3 leads to potentiation of activation by the G-protein beta gamma dimer in an in vitro assay. 1321N1 cells treated with calmodulin inhibitors concurrent with and post stimulation of muscarinic receptors significantly reduced [3H]PIP hydrolysis. Together these data are suggestive of cooperative role for calmodulin in the G protein beta gamma dimer-stimulated activity of PLCbeta3. PMID- 17118345 TI - Glutamate-stimulated peroxynitrite production in a brain-derived endothelial cell line is dependent on N-methyl-D-aspartate (NMDA) receptor activation. AB - There is accumulating and convincing evidence indicating a role for glutamate in the pathogenesis of the human demyelinating disease multiple sclerosis (MS). Studies in experimental autoimmune encephalomyelitis (EAE), the animal model of MS, demonstrate that pharmacological inhibition of specific glutamate receptors suppresses neurological symptoms and prevents blood-brain barrier (BBB) breakdown. The mechanisms through which glutamate influences BBB function during EAE remain unclear. Glutamate triggers the production of nitric oxide and superoxide, which can lead to the formation of peroxynitrite (ONOO(-)). Recent studies have implicated ONOO(-) in the loss of neurovascular integrity during EAE. We propose that glutamate contributes to BBB breakdown via the actions of ONOO(-). The present investigation examined glutamate-induced ONOO(-) formation in the b.End3 brain-derived endothelial cell line. b.End3 cells were incubated with a concentration range of glutamate and ONOO(-) production was assessed over time. Results showed a concentration- and time-dependent increase in ONOO(-) levels in glutamate-treated cells that were suppressed by selective and non selective inhibitors of ONOO(-)-mediated reactions. Specific activation of b.End3 associated NMDA receptors also resulted in a concentration-dependent increase in ONOO(-) production. The ability of b.End3 cells to respond to the presence of glutamate was confirmed through the detection of NMDA receptor immnuoreactivity in cell extracts. In addition, the use of the NMDA receptor antagonists MK-801 and memantine reduced glutamate-mediated ONOO(-) generation from b.End3 cells. The data reinforce the important relationship between glutamate and the NMDA receptor, positioned at neurovascular sites, which may be of particular relevance to the pathogenesis of demyelinating disease. PMID- 17118348 TI - Use of methanesulfonic acid in the reductive ring-opening of O-benzylidene acetals. AB - Methanesulfonic acid was shown to be an efficient and convenient substitute for ethereal HCl in reductive 4,6-O-benzylidene acetal ring-opening reaction with sodium cyanoborohydride in THF. Normal regioselectivity was observed, the 6-O benzyl ethers with free 4-OH group being the major products of the reaction. PMID- 17118347 TI - Normal [3H]flunitrazepam binding to GABAA receptors in the locus coeruleus in major depression and suicide. AB - Major depression and suicide are associated with altered concentrations of specific noradrenergic proteins in the human locus coeruleus (LC). Based on experimental studies that can reproduce these LC abnormalities in laboratory animals, we hypothesized that noradrenergic pathobiology in depression is a result of overactivity of the LC. LC activity is under the control of both excitatory and inhibitory inputs. A major inhibitory input to the LC is GABAergic, arising from the nucleus prepositus hypoglossi. Numerous studies demonstrating low levels of GABA in the CSF and plasma of subjects with major depressive disorder (MDD) raise the possibility that LC overactivity in depression may be secondary to reduced GABAergic input to the LC. Here, GABAergic input to the LC in depression was evaluated by studying the binding of [(3)H]flunitrazepam to GABA(A) receptors at three anatomically defined levels of the human postmortem LC. LC tissues were collected from subjects with MDD, subjects with depressive disorders including MDD that died as a result of suicide, and psychiatrically normal control subjects. A modest rostral-caudal gradient of GABA(A) receptor binding density was observed among all subjects. No significant differences in the amount of binding to GABA(A) receptors were observed between control subjects (n=21) and MDD subjects (n=9) or depressed suicide victims (n=17). These results demonstrate that GABA(A) receptor binding in the LC measured with [(3)H]flunitrazepam is not altered in subjects with depressive illnesses. PMID- 17118349 TI - Acute cysticercosis favours rapid and more severe lesions caused by Leishmania major and Leishmania mexicana infection, a role for alternatively activated macrophages. AB - Parasitic helminths have developed complex mechanisms to modulate host immunity. In the present study we found that previous infection of mice with the cestode Taenia crassiceps favours parasitemia and induces larger cutaneous lesions during both Leishmania major and Leishmania mexicana co-infections. Analysis of cytokine responses into draining lymph nodes indicated that co-infection of T. crassiceps Leishmania did not inhibit IFN-gamma production in response to Leishmania antigens, but significantly increased IL-4 production. Additionally, anti Leishmania-specific IgG1 antibodies and total IgE increased in co-infected mice, whereas, IgG2a titers remained similar. Macrophages from Taenia-infected mice displayed increased mRNA transcripts of arginase-1, Ym1, and Mannose Receptor, as well as greater production of urea (all markers for an alternate activation state) compared to macrophages from Leishmania-infected mice. In contrast, lower mRNA transcripts for IL-12p35, IL-12p40, IL-23p19, and iNOS were detected in macrophages obtained from cestode-infected mice compared to uninfected and Leishmania-infected mice after LPS stimulation. The presence of cestode also generated impaired macrophage anti-leishmanicidal activity in vitro, as evidenced by the inability of these macrophages to prevent Leishmania growth compared to macrophages from uninfected mice. This was observed despite the fact that both groups of cells were exposed to IFN-gamma. Flow cytometry showed high IFN-gammaR expression on Taenia-induced macrophages. Thus, lack of response to IFN-gamma is not associated with the absence of its receptor. Our data suggest that cestode infection may favour Leishmania installation by inducing alternatively activated macrophages rather than inhibiting Th1-type responses. PMID- 17118350 TI - Interaction of photosensitizers with liposomes containing unsaturated lipid. AB - Small unilamellar liposomes were made of dipalmitoyl-phosphatidylcholine and dioleoyl-phosphatidylcholine, and photosensitized by a symmetrically or an asymmetrically substituted glycosilated tetraphenyl-porphyrin derivative. As differential scanning calorimetry and electron paramagnetic resonance spectroscopy (EPR) revealed these porphyrin derivatives were localized in different depth within the lipid bilayer. Both porphyrin derivatives were able to induce photoreaction and consequent structural changes in the membrane. 5-, 12-, or 16-doxyl stearic acid labeled lipid bilayers were applied and the efficiency of photoinduced reaction was followed by the decay of their EPR signal amplitude. Light dose-dependent destruction of nitroxide radical proved to be dependent on the position of spin label. In this process the porphyrin localized in closer connection with the double bond of unsaturated fatty acid was more effective. EPR signal decay was also dependent on the unsaturated fatty acid content of the liposome and the oxygen saturation of the solvent. PMID- 17118351 TI - Low cholesterol solubility in DODAB liposomes. AB - Through the analysis of the ESR spectra of spin labels, we investigated the thermotropic properties of dioctadecyl dimethylammonium bromide (DODAB) liposomes, in low and high ionic strength, with different cholesterol contents. The cationic lipid gel phase is stabilized by the presence of ions, the bilayer having a higher gel/fluid transition temperature (Tm) in high ionic strength. As found for low ionic strength [Benatti, C.R., Feitosa, E., Fernandez, R.M., Lamy Freund, M.T., 2001. Structural and thermal characterization of dioctadecyldimethylammonium bromide dispersions by spin labels. Chem. Phys. Lipids, 111, 93-104], high salt DODAB membranes also present a clear coexistence of the two phases around Tm. Cholesterol solubility in DODAB bilayers seems to be rather low, as the coexistence of DODAB and cholesterol-rich domains can be clearly detected by spin labels, for cholesterol concentration as low as 15 mol% of the total lipid. For lower cholesterol concentrations, the effect of cholesterol in DODAB bilayers is similar to that in phospholipids. For concentrations at or above 45 mol% of cholesterol, spin labels do not detect the coexistence of structurally different domains. PMID- 17118352 TI - Paraoxonase-1 concentrations in end-stage renal disease patients increase after hemodialysis: correlation with low molecular AGE adduct clearance. AB - BACKGROUND: Hemorrhagic stroke and ischemic heart disease continue to be key problems in patients with end stage renal failure. Reduced serum paraoxonase (PON 1) activity has been described in these patients, which could contribute to the accelerated development of atherosclerosis. We hypothesized that retention of uremic toxins and or "middle molecules" including advanced glycation (AGE) free adducts and peptides could play a mechanistic role in decreasing PON-1 activity. METHODS: We enrolled 22 ESRD patients undergoing hemodialysis in whom paired pre- and post-dialysis samples were studied along with 30 age-matched control subjects. RESULTS: ESRD patients showed a 76% decrease in PON-1 activity. As expected, ESRD patients had an increase in lipoperoxides and advanced oxidation protein products (AOPP). Our patients had a 3-fold increase in serum AGEs and a striking 10-fold increase in low molecular weight (<10 kDa) AGEs. Post-dialysis samples in all patients displayed an increase in PON-1 activity, which ranged from 4 to 40% of the predialysis value. HDL-cholesterol, apoAI, free cholesterol (as a LCAT surrogate), HDL-subclasses and TG did not change significantly after dialysis. Changes in PON-1 activity display a good correlation (r=0.66, p<0.001) with rates in which creatinine and urea are cleared. Clearance of low molecular weight AGEs after hemodialysis explains 79% of the changes in PON-1 activity and are hence a much better predictor than creatinine changes (r=0.89, p<0.00). In vitro incubation of paraoxonase with serum ultrafiltrates show a time and concentration dependent inhibition of PON-1 by the ultrafiltrates, an inhibition that is up to 3 times higher (from 8 to 24%) when chronic renal failure patients are the source of the ultrafiltrate. CONCLUSION: We showed that HD results in a significant, consistent increase in the activity of the antioxidant enzyme PON-1. The effect, correlates with the effectiveness of dialysis to clear creatinine and urea, and with the clearance of AGE adducts of low molecular weight. This effect was replicated in vitro, showing time and dose dependency. Our results suggest that another cause for the observed lower PON-1 concentrations in CRF are the retention of low-middle molecules and demonstrate a positive effect of hemodialysis in the delicate oxidant-antioxidant state of these patients, that should be weighted against other pro-oxidant effects that have also been shown to occur previously. If the hypothesis that AGEs are the main culprits is proved in further research, this opens a putative therapeutic avenue for AGE blockers in ESRD. PMID- 17118353 TI - Validation of a new protocol for navigated intraoperative assessment of knee kinematics. AB - BACKGROUND: This study describes a novel method for accurate evaluations of knee kinematics during arthroscopic reconstructions of anterior cruciate ligament (ACL). METHODS: Quantitative evaluation of knee stability was estimated by experimental validation on 30 volunteers and by statistical analysis of test repeatability. RESULTS: Proposed method present short learning time, is minimally invasive and thus suitable for arthroscopic techniques. Computed laxity showed a repeatability of 1.5 degrees for varus-valgus, 3 degrees for internal-external, and 2mm for antero-posterior tests. CONCLUSIONS: This method represents a reliable quantification of knee kinematics in surgery, able to improve present intra-operative assessment of knee stability. PMID- 17118354 TI - Roxithromycin potentiates the effects of chloroquine and mefloquine on multidrug resistant Plasmodium falciparum in vitro. AB - Chloroquine (CQ) and mefloquine (MQ) are no longer potent antimalarial drugs due to the emergence of resistant Plasmodium falciparum. Combination therapy has become the standard for many regimes in overcoming drug resistance. Roxithromycin (ROM), a known p-glycoprotein inhibitor, is reported to have antimalarial activity and it is hoped it will potentiate the effects of both CQ/MQ and reverse CQ/MQ-resistance. We assayed the effects of CQ and MQ individually and in combination with ROM on synchronized P. falciparum (Dd2 strain) cultures. The IC(50) values of CQ and MQ were 60.0+/-5.0 and 16.0+/-3.0 ng/ml; these were decreased substantially when combined with ROM. Isobolograms indicate that CQ-ROM combinations were relatively more synergistic (mean FICI 0.70) than MQ-ROM (mean FICI 0.85) with their synergistic effect at par with CQ-verapamil (VRP) (mean FICI 0.64) and MQ-VRP (mean FICI 0.60) combinations. We conclude that ROM potentiates the CQ/MQ response on multidrug-resistant P. falciparum. PMID- 17118355 TI - Investigating the possible role of benthic macroinvertebrates and zooplankton in the life cycle of the haplosporidian Bonamia ostreae. AB - Bonamia ostreae is a protistan parasite of the European flat oyster, Ostrea edulis. Though direct transmission of the parasite can occur between oysters, it is unclear if this represents the complete life cycle of the parasite, and the role of a secondary or intermediate host or carrier species cannot be ruled out. In this preliminary study, benthic macroinvertebrates and zooplankton from a B. ostreae-endemic area were screened for the presence of parasite DNA, using polymerase chain reaction (PCR). Eight benthic macroinvertebrates and nineteen grouped zooplankton samples gave positive results. Certain species, found positive for the parasite DNA, were then used in laboratory transmission trials, to investigate if they could infect naive oysters. Transmission of B. ostreae was effected to two naive oysters cohabiting with the brittle star, Ophiothrix fragilis. PMID- 17118356 TI - Detection of Leishmania (Viannia) DNA in blood from patients with American cutaneous leishmaniasis. AB - The aim of the present study was to investigate the presence of Leishmania (Viannia) subgenus DNA in peripheral blood from patients with cutaneous lesions due to American cutaneous leishmaniasis. The buffy coats from 68 blood samples were analyzed by polymerase chain reaction using the MP1L/MP3H primers. The parasite DNA was detected in 2 (3.4%) out of 59 patients who had amastigotes present in samples taken from lesions. The presence of Leishmania (Viannia) DNA in the blood of these patients indicates hematogeneous parasite dissemination. PMID- 17118357 TI - Plasmodium falciparum: immunization with MSP1-42 induced non-inhibitory antibodies that have no blocking activities but enhanced the potency of inhibitory anti-MSP1-42 antibodies. AB - Hyperimmunization with Plasmodium falciparum MSP1-42 could induce antibodies that have little or no parasite growth inhibitory activities. These antisera had no blocking activities as determined by their ability to interfere with the in vitro activities of growth inhibitory anti-MSP1-42 sera. Equally important, they enhanced the potency of growth inhibitory anti-MSP1-42 sera. PMID- 17118358 TI - Methylation of Smad6 by protein arginine N-methyltransferase 1. AB - Signal transduction pathways utilize posttranslational modifications to regulate the activity of their components in a temporal-spatial and efficient fashion. Arginine methylation is one of the posttranslational modifications that can result in monomethylated-, asymmetric dimethylated- and/or symmetric dimethylated arginine residues in proteins. Here we demonstrate that inhibitory-Smads (Smad6 and Smad7), but not receptor-regulated- (R-)Smads and the common-partner Smad4, can be methylated by protein arginine N-methyltransferase (PRMT)1. Using mass spectrometric analysis, we found that PRMT1 dimethylates arginine(74) (Arg(74)) in mouse Smad6. PRMT1 interacts with the N-terminal domain of Smad6 in which Arg(74) residue is located. Assays examined so far have shown no significant differences between the functions of Smad6 and those of methylation-defective Smad6 (Smad6R74A). Both wild-type and Smad6R74A were equally efficient in blocking BMP-induced growth arrest upon their ectopic expression in HS-72 mouse B cell hybridoma cells. PMID- 17118359 TI - Distinct mechanisms underlie distinct polyphenol-induced neuroprotection. AB - Glutamate excitotoxicity is mediated by intracellular Ca(2+) overload, caspase-3 activation, and ROS generation. Here, we show that curcumin, tannic acid (TA) and (+)-catechin hydrate (CA) all inhibited glutamate-induced excitotoxicity. Curcumin inhibited PKC activity, and subsequent phosphorylation of NR1 of the NMDA receptor. As a result, glutamate-mediated Ca(2+) influx was reduced. TA attenuated glutamate-mediated Ca(2+) influx only when simultaneously administered, directly interfering with Ca(2+). Both curcumin and TA inhibited glutamate-induced caspase-3 activation. Although Ca(2+) influx was not attenuated by CA, caspase-3 was reduced by direct inhibition of the enzyme. All polyphenols reduced glutamate-induced generation of ROS. PMID- 17118360 TI - Protective effect of acetyl-L-carnitine against oxidative stress induced by antiretroviral drugs. AB - Both HIV infection per se and antiretroviral drugs might contribute to oxidative stress and mitochondrial dysfunctions. In this study we assess zidovudine, stavudine and didanosine on U937 and CEM cell lines. All these drugs induced apoptosis and increased intracellular hydrogen peroxide but not superoxide anions. The addition of acetyl-l-carnitine (ALC) was able to prevent the pro oxidant effect of the drugs tested. Supplementation with ALC, deficient in certain cohorts of HIV-infected individuals, especially on high active antiretroviral therapy regimen, has been associated with favourable effects. These data suggest that one of these effects could be a direct anti-oxidant action. PMID- 17118361 TI - A promoter switch that can rescue a plant sigma factor mutant. AB - Chloroplasts sigma factors act in concert with PEP, the bacterial-type plastid RNA polymerase. Using a sigma knockout line from Arabidopsis thaliana, we investigated mutant-specific changes in plastid gene expression at RNA level. One characteristic feature was the appearance of a long transcript that spans the atpB-E operon and extends considerably into the far-upstream region of atpB. This region reveals a cluster of typical promoter elements for NEP, the second (phage type) plastid RNA polymerase. The NEP promoter cluster can help maintain RNA synthesis in situations where no functional sigma factor is available for PEP. PMID- 17118362 TI - Kinetics, inhibition and oligomerization of Epstein-Barr virus protease. AB - Epstein-Barr virus (EBV) is an omnipresent human virus causing infectious mononucleosis and EBV associated cancers. Its protease is a possible target for antiviral therapy. We studied its dimerization and enzyme kinetics with two enzyme assays based either on the release of paranitroaniline or 7-amino-4 methylcoumarin from labeled pentapeptide (Ac-KLVQA) substrates. The protease is in a monomer-dimer equilibrium where only dimers are active. In absence of citrate the K(d) is 20 microM and drops to 0.2 microM in presence of 0.5M citrate. Citrate increases additionally the activity of the catalytic sites. The inhibitory constants of different substrate derived peptides and alpha-keto-amide based inhibitors, which have at best a K(i) of 4 microM, have also been evaluated. PMID- 17118363 TI - The collapsin response mediator protein 1 (CRMP-1) and the promyelocytic leukemia zinc finger protein (PLZF) bind to UDP-N-acetylglucosamine 2-epimerase/N acetylmannosamine kinase (GNE), the key enzyme of sialic acid biosynthesis. AB - Sialic acids (Sia) are expressed as terminal sugars in many glycoconjugates. They are involved in a variety of cell-cell interactions and therefore play an important role during development and regeneration. UDP-N-acetylglucosamine 2 epimerase/N-acetylmannosamine kinase (GNE) is the key enzyme in the de novo synthesis of Sia and it is a regulator of cell surface sialylation. Inactivation of GNE in mice results in early embryonic lethality. Mutations in the GNE gene are of clinical relevance in hereditary inclusion body myopathy, but these mutations do not necessarily decrease the enzymatic activity of GNE. In this study, we searched for novel function of the GNE protein beside its enzymatic function in the Sia biosynthesis. We here report the identification of novel GNE interacting proteins. Using a human prey matrix we identified four proteins interacting with GNE in a yeast two-hybrid assay. For two of them, the collapsin response mediator protein 1 and the promyelocytic leukemia zinc finger protein, we could verify protein-protein interaction with GNE. PMID- 17118364 TI - Effect of the immunomodulator leflunomide on the induction of endometriosis in an experimental rat model. AB - The effect of immunomodulator leflunomide on the development of an experimental endometriosis model was assessed by surgically transplanting autologous fragments of endometrial tissue onto the inner surface of the abdominal wall and arterial cascades of the small intestines. Leflunomide was found to affect the development of endometriosis negatively and seemed to interfere with the growth and maintenance of the uterine explant in this experimental rat model. PMID- 17118365 TI - Characterization of human sperm populations using conventional parameters, surface ubiquitination, and apoptotic markers. AB - OBJECTIVE: To directly compare distinct assays proposed to monitor human sperm quality and possibly preselect sperm populations for assisted reproductive technology (ART). DESIGN: Analysis of human sperm sample quality using several methodologies. SETTING: Academic and clinical institutions. PATIENT(S): Samples from consenting patients undergoing routine semen analysis or ART. INTERVENTIONS: Human sperm samples were analyzed in terms of World Health Organization parameters and processed for annexin V, terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling of DNA (TUNEL), and the sperm-ubiquitin tag immunoassay (SUTI). Samples were analyzed both by flow cytometry and fluorescence microscopy. MAIN OUTCOME MEASURE(S): Correlations among apoptotic markers (outer leaflet phosphatidylserine exposure, membrane integrity, and DNA fragmentation), external ubiquitination, and semen parameters in human spermatozoa. RESULT(S): Nonviable sperm, TUNEL-positive cells, and ubiquitin fluorescence intensity means inversely correlate with semen parameters. Apoptotic markers do not correlate with sperm surface ubiquitination. Normozoospermic samples have a higher number of viable cells and lower DNA fragmentation compared with samples with abnormal parameters. Nonviable sperm are more prevalent in samples with low counts and poor morphology but not low motility. Not all sperm with morphologic abnormalities present surface ubiquitination. CONCLUSION(S): Sperm quality is inversely correlated with lack of viability, DNA fragmentation, and ubiquitin fluorescence intensity means. However, none of the apoptotic markers correlate with ubiquitin labeling. Elimination of defective sperm cells prior to ART using surface markers (annexin V, ubiquitin) seems unwarranted at this stage. PMID- 17118366 TI - Determining the time androgens and sex hormone-binding globulin take to return to baseline after discontinuation of oral contraceptives in women with polycystic ovary syndrome: a prospective study. AB - In this study, discontinuation of oral contraceptive pills in women with polycystic ovary syndrome was followed by the return of all measured androgens and sex hormone-binding globulin levels to basal values after 8 weeks. These observations are pertinent to the measurement of androgens and sex hormone binding globulin levels in subjects who currently are taking oral contraceptive pills and have symptoms that are related to polycystic ovary syndrome. PMID- 17118367 TI - Use of office hysteroscopy to empty a very large hematometra in a young virgin patient with mosaic Turner's syndrome. AB - OBJECTIVE: To describe the successful management of a hematometra using a 5-mm continuous flow operative office hysteroscope. DESIGN: Case report. SETTING: University of Naples "Federico II." PATIENT(S): A 13-year-old virgin patient affected by mosaic Turner's syndrome (45 X; 46 XX) was referred to the emergency room of the Department of Obstetrics and Gynecology after an episode of severe pelvic pain with metrorrhagia. A large hematometra was detected by transabdominal ultrasound scanning. INTERVENTION(S): Vaginoscopic hysteroscopy performed in outpatient setting. MAIN OUTCOME MEASURE(S): Complete resolution of the hematometra and related clinical symptoms. RESULT(S): Vaginoscopic approach avoided general anesthesia and preserved the integrity of her hymen. A chocolate like fluid started to spill out from the uterine cavity as soon as the tip of hysteroscope passed through the internal uterine ostium. A transabdominal ultrasound performed 2 days later showed resolution of the hematometra. Success of the procedure was confirmed by the resolution of all clinical symptoms. CONCLUSION(S): In selected cases, with intact outflow tract, outpatient vaginoscopic hysteroscopy might represent the therapeutic technique of choice in case of hematometra, even in the case of virgin patients. PMID- 17118368 TI - Blastocyst embryo transfer is associated with a sex-ratio imbalance in favor of male offspring. AB - OBJECTIVE: To evaluate the sex ratio of offspring born after blastocyst transfers. DESIGN: Retrospective data analysis. SETTING: A large assisted reproductive technology center. PATIENT(S): We included 1,284 offspring from 937 deliveries during the period August 2003-August 2005. INTERVENTION(S): Tabulation and statistical analysis of all births resulting from fresh IVF cycles. The sex of resulting offspring was compared in both day 3 and blastocyst transfers for all births and for singleton deliveries. In addition, the sex of children conceived with the use of autologous oocytes and donor oocytes was evaluated. MAIN OUTCOME MEASURE: Sex ratio of offspring born following embryo transfers (ETs) after day 3 of culture and sequential blastocycst culture. RESULT(S): The overall sex ratio was significantly shifted toward males when blastocyst transfers were performed. Blastocyst transfers with only the use of autologous oocytes resulted again in a significantly higher proportion of male offspring. An even greater proportional difference was encountered in singleton offspring from donor oocytes. However, significance was not reached because of the limited number of offspring in the subgroup. CONCLUSION(S): This is the first individual center report of a significant sex-ratio imbalance after the sequential media culture of blastocysts. The large imbalance in singleton births associated with the use of donor oocytes, although not significant, is cautionary in regard to the use of elective single ETs. Observation and publication of phenomena such as the effects of extended culture on the sex ratio of live-borns will allow us a better understanding of early differences in sexual dimorphism of the embryo, and will allow us to counsel our patients more appropriately. PMID- 17118369 TI - Increased type III and V collagen expression in human corpora lutea in early pregnancy. AB - OBJECTIVE: To investigate the possible involvement of collagen in the characteristic structure and function of human corpora lutea (CL), type V collagen expression was determined in the CL tissues during the menstrual cycle and early pregnancy. DESIGN: In vitro experiment. SETTING: Department of Obstetrics and Gynecology, Wakayama Medical University, Wakayama, Japan. PATIENT(S): Regulatory cycling women and pregnant women with ovarian tumor and ectopic pregnancy who underwent adnexectomy. INTERVENTION(S): Composition of the various types of collagen in human CL was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. MAIN OUTCOME MEASURE(S): Expression of type V collagen. RESULT(S): The ratios of type III to type I collagen and the ratios of type V to type I collagen in the CL tissues were significantly increased in early pregnancy compared with those in the menstrual cycle. CONCLUSION(S): These results suggest that alterations in composition of collagen might play an important role in determining the physiology and structure of the CL during the menstrual cycle and early pregnancy. PMID- 17118370 TI - Newly developed reconstituted high-density lipoprotein containing sphingosine-1 phosphate induces endothelial tube formation. AB - Reconstituted high-density lipoprotein (rHDL) has been shown to produce a rapid regression of atherosclerosis in animal models and humans. Sphingosine-1 phosphate (S1P), which is a bioactive lipid in HDL, plays a role in mitogenesis, endothelial cell motility, and cell survival, as well as organization and differentiation into a vessel. In this study, we examined the direct role of a newly developed rHDL, [POPC(1-palmitoyl-2-oleoyl phosphatidylcholine)/S1P/apolipoproteinA-I(A-I)]rHDL containing S1P in tube formation in endothelial cells (ECs) as well as cholesterol efflux in macrophage. The effect of (POPC/S1P/A-I)rHDL on cholesterol efflux in macrophage was similar to that of conventional rHDL, (POPC/A-I)rHDL. In addition, (POPC/S1P/A-I)rHDL induced EC proliferation through the activation of phospho-Akt and phospho extracellular-signal-regulated kinases (p-ERK) 1/2 and EC tube formation, and this effect was blocked by inhibitors of Akt, ERK and endothelial nitric-oxide synthase (eNOS). In addition, (POPC/S1P/A-I)rHDL-induced p-ERK1/2 activation and EC tube formation can be mainly attributed to S1P-stimulated signaling through S1P2 and S1P3 as determined by an anti-sense strategy. In conclusion, (POPC/S1P/A I)rHDL induces cholesterol efflux independently of S1P but has additional S1P mediated effects on EC tube formation mediated by Akt/ERK/NO through S1P2 and S1P3. In the future, these new discs may be useful for the treatment of atherosclerotic and ischemic cardiovascular disease, such as acute coronary syndrome and atherosclerosis obliterans. PMID- 17118371 TI - Age-related increases in circulating inflammatory markers in men are independent of BMI, blood pressure and blood lipid concentrations. AB - OBJECTIVE: To examine whether age-related increase in concentrations of circulating inflammatory mediators is due to concurrent increases in cardiovascular risk factors or is independent of these. METHODS AND RESULTS: Cytokines (IL-6, IL-18), chemokines (6Ckine, MCP-1, IP-10), soluble adhesion molecules (sICAM-1, sVCAM-1, sE-selectin) and adipokines (adiponectin) were measured in the plasma of healthy male subjects aged 18-84 years (n=162). These were related to known cardiovascular risk factors (age, BMI, systolic and diastolic blood pressure, plasma total cholesterol, LDL cholesterol, HDL cholesterol and triacylglycerol concentrations) in order to identify significant associations. Plasma concentrations of sVCAM-1, sE-selectin, IL-6, IL-18, MCP-1, 6Ckine, IP-10 and adiponectin, but not sICAM-1, were significantly positively correlated with age, as well as with several other cardiovascular risk factors. The correlations with other risk factors disappeared when age was controlled for. In contrast, the correlations with age remained significant for sVCAM-1, IL-6, MCP-1, 6Ckine and IP-10 when other cardiovascular risk factors were controlled for. CONCLUSIONS: Plasma concentrations of some inflammatory markers (sVCAM-1, IL 6, MCP-1, 6Ckine, IP-10) are positively correlated with age, independent of other cardiovascular risk factors. This suggests that age-related inflammation may not be driven by recognised risk factors. PMID- 17118372 TI - Renin-angiotensin system gene polymorphisms and coronary artery disease in a large angiographic cohort: detection of high order gene-gene interaction. AB - There have been many reports regarding the association between renin-angiotensin system (RAS) gene polymorphisms and coronary artery disease (CAD) or acute myocardial infarction (AMI), but the results are inconsistent. In the present study, we used several new approaches with multilocus data to reappraise this issue in a large and relatively homogeneous Taiwanese population. A total of 1254 consecutive patients who underwent cardiac catheterization (735 with documented coronary artery disease and 519 without) between 1996 and 2003 were recruited. Angiotensin-converting enzyme gene insertion/deletion (I/D) polymorphism; T174M, M235T, G-6A, A-20C, G-152A and G-217A polymorphisms of the angiotensinogen gene; and A1166C polymorphism of the angiotensin II type I receptor gene were genotyped. In single-locus analyses, no locus was associated with CAD, history of AMI and three-vessel CAD, either with or without adjustment for conventional CAD risk factors. For multilocus analyses, we recreated a balanced population, with the controls individually matched to the cases regarding the conventional CAD risk factors. We found that the angiotensinogen gene haplotype profile was significantly different between the cases and controls (chi2=31.6, P=0.030) in haplotype analyses. Furthermore, significant three-locus (G-217A, M235T and I/D) gene-gene interactions were detected by multifactor-dimensionality reduction method (highest cross-validation consistency 10.0, lowest prediction error 40.56%, P=0.017) and many even higher order gene-gene interactions by multilocus genotype disequilibrium tests (16 genotype disequilibria exclusively found in the controls, all of which included at least two genes among AGT, ACE and AT1R genes). Our study is the first to demonstrate epistatic, high-order, gene-gene interactions between RAS gene polymorphisms and CAD. These results are compatible with the concept of multilocus and multi-gene effects in complex diseases that would be missed with conventional approaches. PMID- 17118373 TI - Temporal shifting: a hidden key to the skewed peak puzzle. AB - The recorder-provided peak position for flow-type chemical instruments has been verified mathematically as being comprised of a "spatially-non-existent" shift, which is generated due to the relativity in accounting for the detection at a fixed point. This shift, denoted as Phi, can be approximated by Phi approximately 0.5micro(t)2, where micro(t) is the temporal expanding coefficient of the system given. For flow injection analysis, the shift is correlated to a longitudinal dispersion coefficient D and the flow speed u, i.e., Phi approximately D/u2. For linear chromatography, it is correlated to a dynamic partition ratio k'' and a scaling factor f of the column used, i.e., Phi approximately 0.5k''f. In combination, the temporal shift can be expressed as Phi approximately 0.5k''f+D(k''+1)2/u2. Although the shift may be small in scale, it provides a clue to decipher the basic parameters from a recorded peak. Under a linear isotherm, this parameter can be estimated readily from an experimental peak following a very simple procedure. PMID- 17118374 TI - Sensitive and convenient method for the quantification of clonidine in serum of pediatric patients using liquid chromatography/tandem mass spectrometry. AB - An improved and easy to use liquid chromatography/tandem mass spectrometric (LC/MS/MS) method in human serum was developed for the quantification of clonidine (CLD), an alpha2-/alpha1-adrenoceptor agonist, used for analgo-sedation and the therapy of opioid withdrawal in pediatric patients. Sample preparation consisted of precipitation of serum proteins by adding acetonitrile and centrifugation of the sample subsequently. [(2)H4]Clonidine (CLD4) served as internal standard. Chromatographic separation of the supernatant was achieved using a 100mmx3mm, 5microm Thermo Electron BetaBasic C4 column with isocratic flow and elution consisting of 0.1% formic acid/acetonitrile (85/15, v/v) and a flow-rate of 350microl/min resulting in a column pressure of 280-420kPa. LC/MS/MS detection was performed by using a triple-stage quadrupole mass spectrometer (TSQ Quantum, Thermo Electron) working in selected reaction monitoring mode with positive electrospray ionization. The analyte was quantified in a single run within 5min. Linearity was demonstrated over the expected concentration range 0.15-50microg/l CLD. The lower limit of quantification (LLOQ) and the limit of detection were 0.1microg/l and 0.01microg/l, respectively. None of the drugs used concomitantly during analgesic therapy interfered in the assay in vitro. Intra day precision expressed as RSD was 9.6% or less for CLD, while inter-day result was 10.0% or less for CLD. Intra-day and inter-day accuracy was within +/-4.9% and +/-1.8%, respectively. The method was validated according to the international guidelines of the International Conference on Harmonisation (ICH) and the US Food and Drugs Administration (FDA). The described method is suitable to analyse serum samples with very small volumes and sets the stage for pharmacokinetic studies in pediatric studies. PMID- 17118375 TI - Development of an algorithm for peak detection in comprehensive two-dimensional chromatography. AB - A method for peak detection in two-dimensional chromatography is presented. The algorithm applies first the methods developed for peak detection in one dimensional chromatography to detect peaks in one dimension. In a second step, a decision tree is applied to decide which one-dimensional peaks are originated from the same compound and have to be 'merged' into one two-dimensional peak. To this end, different features of the peaks (second-dimension peak regions and second-dimension retention times) are compared and different criteria (common peak regions, retention time differences, unimodality in the first dimension) are applied. Different options can be used, depending on the nature of the data. The user controls this decision tree by establishing several options and "switches". The algorithm was tested with GCxGC chromatograms obtained for a commercial air freshener sample, detecting and merging the modulated peaks belonging to the same compound. Recommendations for the set of options and switches are given. A utility that calculates and sums peak areas from merged peaks is added to facilitate automated quantification. Although the algorithm was developed for GCxGC, its application to comprehensive two-dimensional liquid chromatography (LCxLC) data should at most require minor modifications. PMID- 17118376 TI - On-site polymer-coated hollow fiber membrane microextraction and gas chromatography--mass spectrometry of polychlorinated biphenyls and polybrominated diphenyl ethers. AB - Porous polypropylene hollow fiber membrane coated with a conjugated polymer was used as an on-site sampling device for the extraction of polychlorinated biphenyls and polybrominated biphenyl ethers from coastal sea water samples. The coated hollow fiber membrane was placed in a vial containing the sample, and the target compounds extracted via manual shaking of the vials at the site of sample collection. For each extraction, two fibers were used. After extraction, the fibers with the adsorbed analytes were brought back to the laboratory for further processing. Care was taken to preserve the integrity of the analytes and to avoid contamination during transport; after extraction, the fibers were carefully removed and placed in air-tight crimper vials which were stored in an ice-box. The analytes were desorbed by solvent in the laboratory and analyses were carried out using gas chromatography/mass spectrometry. This method was highly reproducible with relative standard deviations in the range of 1-9%. Recoveries from spiked water samples ranged from 83% to 98%. Low limits of detections between 0.04 and 0.21ngl(-1) were achieved. The extraction efficiency was compared with solid-phase microextraction. PMID- 17118377 TI - Simultaneous determination of amphoteric surfactants in detergents by capillary electrophoresis with indirect UV detection. AB - A simple, rapid and simultaneous determination of four types of amphoteric surfactants, i.e., C8, C10, C12, C14, C16 and C18-homologues of alkyldimethylamine N-oxide (AO), alkylamidopropylamine N-oxide (APAO), alkylbetaine (Bt) and alkylamidopropylbetaine (APB), was performed by using capillary electrophoresis (CE) with indirect UV detection. To optimize the separation conditions, effects of pH of background solution (BGS), organic modifier and chromophore for indirect UV detection on the CE separation of the amphoteric compounds were investigated. Addition of 50% (v/v) acetonitrile to the BGS under a lower pH condition brought a good separation performance due to the suppression of micelle formation for the analytes and the adsorption onto the inner surface of the capillary. Under an optimal condition, the 24 amphoteric analytes were completely separated in a single run within 17min. The relative standard deviation of the migration time was ranging from 0.20 to 0.23% and the limit of detection values for AO, APAO, Bt and APB homologues were 10-20, 20, 20 50 and 50microg/mL, respectively. Furthermore, the developed method can provide a high resolution separation of the amphoteric surfactants in commercially available detergents and shampoo without any sample pretreatments. PMID- 17118378 TI - Retention of ionisable compounds on high-performance liquid chromatography XVII. Estimation of the pH variation of aqueous buffers with the change of the methanol fraction of the mobile phase. AB - The use of methanol-aqueous buffer mobile phases in HPLC is a common election when performing chromatographic separations of ionisable analytes. The addition of methanol to the aqueous buffer to prepare such a mobile phase changes the buffer capacity and the pH of the solution. In the present work, the variation of these buffer properties is studied for acetic acid-acetate, phosphoric acid dihydrogenphosphate-hydrogenphosphate, citric acid-dihydrogencitrate hydrogencitrate-citrate, and ammonium-ammonia buffers. It is well established that the pH change of the buffers depends on the initial concentration and aqueous pH of the buffer, on the percentage of methanol added, and on the particular buffer used. The proposed equations allow the pH estimation of methanol-water buffered mobile phases up to 80% in volume of organic modifier from initial aqueous buffer pH and buffer concentration (before adding methanol) between 0.001 and 0.01 mol L(-1). From both the estimated pH values of the mobile phase and the estimated pKa of the ionisable analytes, it is possible to predict the degree of ionisation of the analytes and therefore, the interpretation of acid-base analytes behaviour in a particular methanol-water buffered mobile phase. PMID- 17118379 TI - Determination of phenolic compounds in grape skin by capillary electrophoresis with simultaneous dual fluorescence and diode array absorption detection after dynamic superheated liquid leaching. AB - A fast method for the analysis of 10 of the characteristic compounds of the phenolic fraction in grape skin is proposed here. The method is based on a leaching step by superheated ethanol-water at 120 degrees C and 80bar, which enables to maintain the leachant in liquid state and wide the range of leachable compounds (polar, mid-polar and relatively non-polar compounds) by decrease of the dielectric constant; thus, allowing high leaching efficiencies to be achieved in 30min. After leaching, the target analytes were separated by capillary electrophoresis using a 50mM sodium tetraborate with 10% methanol (pH 8.4) solution as background electrolyte. Determination was performed by simultaneous dual diode array absorption and fluorescence detection, the combination of which increased the selectivity of the overall method, particularly interesting taking into account the complexity of the leachate as no additional concentration and/or clean-up steps were required prior to electrophoretic separation, which lasted only 10min. The short time of the electrophoretic step makes it useful as a screening tool of the target analytes in commercial and non-commercial extracts belonging to the pharmaceutical, cosmetic, nutraceutical or food fields. PMID- 17118380 TI - Effect of salt on purification of plasmid DNA using size-exclusion chromatography. AB - In the present study, we compared the performances of size-exclusion chromatography for the purification of plasmid DNA when different concentrations (0.5M, 1M, 2M, respectively) of two types of salt (NaCl and (NH(4))(2)SO(4)) are present in running buffers. Our experiment results displayed that it is not only the resolution of RNA but also those of supercoiled plasmid DNA and host's genomic DNA were increased greatly in the presence of high concentration of water structure salt. We deduce that two separation modes may be involved in the process: The supercoiled plasmid DNA is influenced mainly by compaction effect and eluted in the size-exclusion mode; whereas, RNA and genomic DNA are influenced mainly by hydrophobic effect due to their stretched and loose structures and eluted in the interaction mode. This method led to an improved efficiency of size-exclusion chromatography. PMID- 17118381 TI - Acidic herbicides in surface waters of Lower Fraser Valley, British Columbia, Canada. AB - In the period 2003-2005 a study was conducted to determine the occurrence, spatial and temporal distribution of five acidic herbicides in the Lower Fraser Valley (LFV) region of British Columbia, Canada. A high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS) method capable of detecting analytes at the sub ng/L level was developed for this study. Samples were collected and analyzed from two references, five agricultural, two urban and five agricultural and urban mixed sites. Only (4-chloro-2-methylphenoxy)acetic acid and triclopyr were detected at the reference sites. The highest concentration of herbicide detected at the reference sites was 0.109ng/L for (4 chloro-2-methylphenoxy)acetic acid. Varying levels of all of the herbicides monitored were detected at the urban, agricultural and the mixed sites. For the urban sites the highest concentration of herbicide detected was 66.6ng/L for 2-(4 chloro-2-methylphenoxy)propanoic acid. For the agricultural sites the highest concentration of herbicide detected was 345ng/L for (2,4-dichlorophenoxy)acetic acid (2,4-D). For the mixed sites the highest concentration of herbicide detected was 1230ng/L for 2,4-D. Overall the mixed sites showed highest concentrations and detection frequencies followed by the agricultural and urban sites. With few exceptions higher concentrations of herbicides were observed for samples collected during spring than for samples collected during fall. The detected concentrations of herbicides were evaluated against established water quality criteria. Herbicide data presented in this study provide reference levels for future pesticide monitoring programs in the region. PMID- 17118382 TI - The dispersion-stability diagram of boehmite nanoparticles in aqueous AOT solutions. AB - The dispersion stability of hydrophilic boehmite nanoparticles in aqueous sodium bis(2-ethylhexyl) sulfosuccinate (AOT) solutions was studied in a wide range of particle and surfactant concentrations. The two experimental parameters significantly influence the dispersion stability and span a stability diagram. With increasing surfactant concentration and decreasing particle concentration, the system changes from stable via moderately stable to unstable and back. In concentrated AOT solutions, fully redispersed particles are present, stabilized by a surfactant bilayer or admicelles on the surface. The redispersion can be reversibly induced by dilution or concentration of the samples. The positions of two transitions, namely for complete precipitation and for beginning redispersion, can be fitted accurately using a simple model based on H-type adsorption and including the specific surface area of the particle and the molar area of the surfactant. The transitions are controlled by the concentration of free surfactant molecules in solution as well as the saturation surface coverage and were corroborated by turbidity measurements. PMID- 17118383 TI - Synthesis of yttrium iron garnet using polymer-metal chelate precursor. AB - Two kinds of polymer-metal compounds, heterogeneous complexes (metal-chelating copolymer microspheres, MCP) and homogeneous complexes (water-soluble metal chelating polymers, WSMCP), were synthesized to act as nucleation agents for YIG precursor preparation in this text. Both of the metal-chelating polymers have the same chelating group and high metal ion adsorption ability from the FTIR and ICP measurement. Furthermore, good YIG crystals can be obtained by treating the MCP precursor with a low calcination temperature at 600 degrees C from the XRD spectra and TEM micrograph. However, the YIG crystal obtained using a WSMCP precursor should be synthesized at a higher calcination temperature (>900 degrees C) due to the different components of the YIG precursor. In addition, the YIG crystal obtained by using the MCP precursor had nearly superparamagnetic behavior after VSM examination. PMID- 17118384 TI - Energy transfer between rhodamine 3B and oxazine 4 in synthetic-saponite dispersions and films. AB - The objective of this study was the investigation of energy transfer between the laser dyes rhodamine 3B (R3B) and oxazine 4 (Ox4) adsorbed on the surface of synthetic Sumecton saponite (Sum). The process of energy transfer was studied for both saponite dispersions and oriented solid films. The electronic properties, luminescence, and the energy transfer process were described by UV-vis absorption and fluorescence spectroscopy. For the efficiency of the energy transfer process, the concentrations of energy donor and acceptor components on a clay mineral surface were found to be essential. A side reaction of the molecular assembly formation reduced both the luminescence and energy-transfer yields, mainly due to fluorescence quenching. The quenching was more problematic for the solid film specimens, where an appropriate modification of the inorganic host with hydrophobic alkylammonium cations was used to achieve a higher luminescence. Due to the higher tendency of Ox4 to form nonluminescent aggregates at higher concentrations, the lowering of the Ox4 concentration further improved the luminescent properties of the films. In this case, the energy transfer occurring in the solid film from R3B to Ox4 was clearly proven. PMID- 17118385 TI - Time-dependence of pervaporation performance for the separation of ethanol/water mixtures through poly(vinyl alcohol) membrane. AB - To clarify the cause of time-dependent separation behavior, the pervaporation performance with operating time through pure poly(vinyl alcohol) (PVA) membrane and glutaraldehyde (GA) cross-linked PVA membranes was investigated. The results showed that the water concentration in the permeate for the air-side surface of the PVA membrane increased dramatically from 92.2 to 95.7% in about 110 min and then remained almost unchanged. However, the water selectivity for the glass-side surface did not change with operating time. Similar results were observed for the GA cross-linked PVA membranes. Furthermore, the contact angle of water on the air side surfaces of those membranes decreased with the time of contact with the feed. These results revealed that this dynamic pervaporation process was mainly attributable to the reconstruction of hydroxyl groups at the air-side surfaces of PVA membranes in response to the change of their surrounding medium during pervaporation. The reconstruction at the glass-side surface of the membrane did not occur because of the preferential localization of hydroxyl groups at the interface between the membrane and the glass plate during film formation of PVA solution. The above conclusion was further confirmed by the following results. The water concentration in the permeate through PVA membranes with the air-side surface facing the feed reached equilibrium more quickly with increasing operation temperature or decreasing degree of cross-linking, which was consistent with the fact that the rate of surface reconstruction accelerated with the increase of temperature or the decrease of the degree of cross-linking. PMID- 17118386 TI - Effect of ionic strength on the rheological behavior of aqueous cetyltrimethylammonium p-toluene sulfonate solutions. AB - The influence of ionic environment on the rheological properties of aqueous cetyltrimethylammonium p-toluene sulfonate (CTAT) solutions has been studied under three different flow fields: simple shear, opposed-jets flow and porous media flow. Emphasis was placed in the experiments on a range of CTAT concentration in which wormlike micelles were formed. It is known that these solutions exhibit shear thickening in the semi-dilute regime, which has been explained in terms of the formation of shear-induced, cooperative structures involving wormlike micelles. In simple shear flow, the zero shear viscosity exhibits first an increase with salt addition followed by a decrease, while the critical shear rate for shear thickening increases sharply at low salt contents and tends to saturate at relatively high ionic strengths. The results are explained in terms of a competition between micellar growth induced by salt addition and changes in micellar flexibility caused by ionic screening effects. Dynamic light scattering results indicate that micelles grow rapidly upon salt addition but eventually achieve a constant size under static conditions. These observations suggest that the wormlike micelles continuously grow with salt addition, but, as they become more flexible due to electrostatic screening, the wormlike coils tend to adopt a more compact conformation. The trends observed in the apparent viscosities measured in porous media flows seem to confirm these hypotheses-but viscosity increases in the shear thickening region-and are magnified by micelle deformation induced by the elongational nature of the local flow in the pores. In opposed-jets flow, the solutions have a behavior that is close to Newtonian, which suggests that the range of strain rates employed makes the flow strong enough to destroy or prevent the formation of cooperative micellar structures. PMID- 17118387 TI - Modeling synergistic adsorption of phenol/aniline mixtures in the aqueous phase onto porous polymer adsorbents. AB - The adsorption equilibria of phenol and aniline on nonpolar polymer adsorbents (NDA-100, XAD-4, NDA-16 and NDA-1800) were investigated in single- and binary solute adsorption systems at 313 K. The results showed that all the adsorption isotherms of phenol and aniline on these adsorbents can be well fitted by Freundlich and Langmuir equations, and the experimental uptake of phenol and aniline in all binary-component systems is obviously higher than predicted by the extended Langmuir model, arising presumably from the synergistic effect caused by the laterally acid-base interaction between the adsorbed phenol and aniline molecules. A new model (MELM) was developed to quantitatively describe the synergistic adsorption behavior of phenol/aniline equimolar mixtures in the binary-solute systems and showed a marked improvement in correlating the binary solute adsorption of phenol and aniline by comparison with the widely used extended Langmuir model. The newly developed model confirms that the synergistic coefficient of one adsorbate is linearly correlated with the adsorbed amount of the other, and the larger average pore size of adsorbent results in the greater synergistic effect of phenol/aniline equimolar mixtures adsorption. PMID- 17118388 TI - An investigation of language and phonological development and the responsiveness of preschool age children to the Lidcombe Program. AB - Knowledge of variables that predict treatment time is of benefit in deciding when to start treatment for early stuttering. To date, the only variable clearly related to treatment time with the Lidcombe Program is pre-treatment stuttering frequency. Previous studies have shown that children whose stuttering is more severe take longer to complete Stage 1 of the program. However, studies to date have not investigated phonology and language as predictors of treatment time. In the context of a Phase II clinical trial, the present prospective study showed that phonological development does not predict treatment time but that, together, stuttering severity, MLU and CELF Receptive Score predict 35-45% of the variance for time taken to complete Stage 1. LEARNING OUTCOMES: The reader should be able to (1) understand guidelines developed for the timing of intervention with the Lidcombe Program based on previous retrospective studies, (2) determine whether pre-treatment language and phonological development play a role in treatment-led recovery with the Lidcombe Program and (3) understand recent empirical evidence on time taken by preschool children to complete Stage 1 of the Lidcombe Program. PMID- 17118392 TI - The value of different electrocardiographic depolarization criteria in the diagnosis of arrhythmogenic right ventricular dysplasia/cardiomyopathy. AB - BACKGROUND: The use of electrocardiographic (ECG) depolarization and repolarization criteria plays a large role in the diagnosis of arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C). Different ECG algorithms should be analyzed in making the diagnosis of ARVD/C with the use of normal and modified recording techniques. METHODS: In a cohort of 343 patients (210 men and 133 women; mean age, 46.0 +/- 13.7 years) meeting the Task Force of the Working Group on Myocardial and Pericardial Diseases of the European Society of Cardiology and the Scientific Council on Cardiomyopathies of the International Society and Federation of Cardiology diagnostic criteria for ARVD/C, the value of different ECG criteria (eg, localized right precordial QRS prolongation defined as QRS duration in (V1+V2+V3)/(V4+V5+V6) of 1.2 or higher, right precordial QRS prolongation with QRS in V1-3 of 110 milliseconds or higher, epsilon potentials in the right precordial leads, S-wave upstroke in V1-3 of 55 milliseconds or higher, and right precordial T-wave inversions) was analyzed with the use of a normal recording technique and a highly amplified and modified recording technique (n = 207) at a paper speed of 50 mm/s. Fifty-two phenotypically and genotypically unaffected individuals identified by systematic screening in 24 families (30 men; mean age, 42.4 +/- 8.3 years) were treated as control subjects. RESULTS: In the normal as well as highly amplified and modified recording techniques, the incidence of localized right precordial QRS prolongation was 98% (100%), that of QRS in V1-3 of 110 milliseconds or higher was 75% (80%), that of prolonged right precordial S-wave upstroke was 84% (60%), that of epsilon potentials was 23% (77%), and that of right precordial T-wave inversions was 55%. Four of 6 patients without the phenomenon of localized right precordial QRS prolongation with the use of the normal recording technique had a prolonged S wave upstroke of 55 milliseconds or higher. In the control group, localized right precordial QRS prolongation, QRS in V1-3 of 110 milliseconds or higher, and epsilon potentials could not be identified. An S-wave upstroke of 55 milliseconds or higher was present in 2 of 3 cases, and T-wave inversions were found in 3. CONCLUSIONS: Electrocardiographic depolarization criteria for ARVD/C analyzed in this large cohort of patients meeting the International Society and Federation of Cardiology/European Society of Cardiology criteria presented with high sensitivity and specificity in comparison with those in the control group of phenotypically and genotypically unaffected individuals defined by systematic screening in 24 families with ARVD/C. The incidence of right precordial T-wave inversions was much lower, indicating that not only patients with overt right ventricular dilatation and dysfunction were included. Electrocardiographic algorithms, including localized right precordial QRS prolongation, prolonged S wave upstroke, and epsilon potentials, with the use of the normal recording technique and the amplified and modified recording technique at a paper speed of 50 mm/s contribute significantly to the noninvasive diagnosis of ARVD/C. PMID- 17118393 TI - Precordial S-T segment elevation. PMID- 17118395 TI - Computerized serial comparison of electrocardiograms: program performance in myocardial infarction. AB - Serial comparison of electrocardiograms (ECGs) can provide a useful clinical function by reporting to the editing cardiologist the diagnostic changes that have occurred since the previous ECG. This program detects "significant measurement differences" in each of the diagnostic categories to detect these changes. We evaluated the accuracy and use of this serial comparison program by comparing the diagnostic results of the program with those of an expert cardiologist using a database of ECGs obtained from patients with symptoms admitted to the hospital and other laboratory results consistent with acute myocardial infarction. We found that the level of agreement between the computer and the cardiologist was much higher when a current ECG was compared with a previous that had been edited by the cardiologist than when that same ECG was analyzed in isolation. PMID- 17118397 TI - Proceedings and abstracts of the Annual International Society of Electrocardiology Meeting, 33rd International Congress on Electrocardiology and the 47th International Symposium on Vectorcardiography, 2006. PMID- 17118398 TI - Working memory deficit in children with mathematical difficulties: a general or specific deficit? AB - This study examined whether children with mathematical difficulties (MDs) or comorbid mathematical and reading difficulties have a working memory deficit and whether the hypothesized working memory deficit includes the whole working memory system or only specific components. In the study, 31 10-year-olds with MDs and 37 10-year-olds with both mathematical and reading difficulties were compared with 47 age-matched and 50 younger controls (9-year-olds) on a number of working memory tasks. Compared with the age-matched controls, both groups of children with MDs performed worse on tasks tapping the central executive (e.g., visual matrix span) and the phonological loop (e.g., word span). More important, the MD group performed worse than the younger controls on the counting span task, whereas the group with comorbid mathematical and reading difficulties performed worse on the counting span task and the visual matrix span task. These findings provide support for the assumption that children with MDs have a working memory deficit. More specifically, children with MDs have a central executive deficit connected to concurrent processing and storage of numerical and visual information. PMID- 17118399 TI - Programmed cell death in flight muscle histolysis of the house cricket. AB - We have characterized the process of flight muscle histolysis in the female house cricket, Acheta domesticus, through analysis of alterations of tissue wet weight, total protein content, and percent shortening of the dorsal longitudinal flight muscles (DLMs). Our objectives were to (1) define the normal course of histolysis in the cricket, (2) analyze the effects of juvenile hormone (JH) removal and replacement, (3) determine the effects of cycloheximide treatment, and (4) examine patterns of protein expression during histolysis. Our results suggest that flight muscle histolysis in the house cricket is an example of an active, developmentally regulated cell death program induced by an endocrine signal. Initial declines of total protein in DLMs indicated the JH signal that induced histolysis occurred by Day 2 and that histolysis was essentially complete by Day 3. Significant reductions in tissue weight and percent muscle shortening were observed in DLMs from Day 3 crickets. Cervical ligation of Day 1 crickets prevented histolysis but this inhibition could be reversed by continual topical treatments with methoprene (an active JH analog) although ligation of Day 2 crickets did not prevent histolysis. A requirement for active protein expression was demonstrated by analysis of synthesis block by cycloheximide and short-term incorporation of (35)S-methionine. Treatment with cycloheximide prevented histolysis. Autofluorographic imaging of DLM proteins separated by electrophoresis revealed apparent coordinated regulation of protein expression. PMID- 17118400 TI - Structural transitions and thermodynamics of a glycine-dependent riboswitch from Vibrio cholerae. AB - Riboswitches are complex folded RNA domains found in noncoding regions of mRNA that regulate gene expression upon small molecule binding. Recently, Breaker and coworkers reported a tandem aptamer riboswitch (VCI-II) that binds glycine cooperatively. Here, we use hydroxyl radical footprinting and small-angle X-ray scattering (SAXS) to study the conformations of this tandem aptamer as a function of Mg(2+) and glycine concentration. We fit a simple three-state thermodynamic model that describes the energetic coupling between magnesium-induced folding and glycine binding. Furthermore, we characterize the structural conformations of each of the three states: In low salt with no magnesium present, the VCI-II construct has an extended overall conformation, presumably representing unfolded structures. Addition of millimolar concentrations of Mg(2+) in the absence of glycine leads to a significant compaction and partial folding as judged by hydroxyl radical protections. In the presence of millimolar Mg(2+) concentrations, the tandem aptamer binds glycine cooperatively. The glycine binding transition involves a further compaction, additional tertiary packing interactions and further uptake of magnesium ions relative to the state in high Mg(2+) but no glycine. Employing density reconstruction algorithms, we obtain low resolution 3-D structures for all three states from the SAXS measurements. These data provide a first glimpse into the structural conformations of the VCI-II aptamer, establish rigorous constraints for further modeling, and provide a framework for future mechanistic studies. PMID- 17118401 TI - The conformations of the manganese transport regulator of Bacillus subtilis in its metal-free state. AB - The manganese transport regulator (MntR) from Bacillus subtilis binds cognate DNA sequences in response to elevated manganese concentrations. MntR functions as a homodimer that binds two manganese ions per subunit. Metal binding takes place at the interface of the two domains that comprise each MntR subunit: an N-terminal DNA-binding domain and a C-terminal dimerization domain. In order to elucidate the link between metal binding and activation, a crystallographic study of MntR in its metal-free state has been undertaken. Here we describe the structures of the native protein and a selenomethionine-containing variant, solved to 2.8 A. The two structures contain five crystallographically unique subunits of MntR, providing diverse views of the metal-free protein. In apo-MntR, as in the manganese complex, the dimer is formed by dyad-related C-terminal domains that provide a conserved structural core. Similarly, each DNA-binding domain largely retains the folded conformation found in metal bound forms of MntR. However, compared to metal-activated MntR, the DNA-binding domains move substantially with respect to the dimer interface in apo-MntR. Overlays of multiple apo-MntR structures indicate that there is a greater range of positioning allowed between N and C-terminal domains in the metal-free state and that the DNA-binding domains of the dimer are farther apart than in the activated complex. To further investigate the conformation of the DNA-binding domain of apo-MntR, a site directed spin labeling experiment was performed on a mutant of MntR containing cysteine at residue 6. Consistent with the crystallographic results, EPR spectra of the spin-labeled mutant indicate that tertiary structure is conserved in the presence or absence of bound metals, though slightly greater flexibility is present in inactive forms of MntR. PMID- 17118402 TI - Crystal structure analysis and solution studies of human Lck-SH3; zinc-induced homodimerization competes with the binding of proline-rich motifs. AB - In cytosolic Src-type tyrosine kinases the Src-type homology 3 (SH3) domain binds to an internal proline-rich motif and the presence or the absence of this interaction modulates the kinase enzymatic activity. The Src-type kinase Lck plays an important role during T-cell activation and development, since it phosphorylates the T-cell antigen receptor in an early step of the activation pathway. We have determined the crystal structure of the SH3 domain from Lck kinase at a near-atomic resolution of 1.0 A. Unexpectedly, the Lck-SH3 domain forms a symmetrical homodimer in the crystal and the dimer comprises two identical zinc-binding sites in the interface. The atomic interactions formed across the dimer interface resemble strikingly those observed between SH3 domains and their canonical proline-rich ligands, since almost identical residues participate in both contacts. Ultracentrifugation experiments confirm that in the presence of zinc ions, the Lck-SH3 domain also forms dimers in solution. The Zn(2+) dissociation constant from the Lck-SH3 dimer is estimated to be lower than 100 nM. Moreover, upon addition of a proline-rich peptide with a sequence corresponding to the recognition segment of the herpesviral regulatory protein Tip, competition between zinc-induced homodimerization and binding of the peptide can be detected by both fluorescence spectroscopy and analytical ultracentrifugation. These results suggest that in vivo, too, competition between Lck-SH3 homodimerization and binding of regulatory proline-rich sequence motifs possibly represents a novel mechanism by which kinase activity is modulated. Because the residues that form the zinc-binding site are highly conserved among Lck orthologues but not in other Src-type kinases, the mechanism might be peculiar to Lck and to its role in the initial steps of T-cell activation. PMID- 17118403 TI - HURP wraps microtubule ends with an additional tubulin sheet that has a novel conformation of tubulin. AB - HURP is a newly discovered microtubule-associated protein (MAP) required for correct spindle formation both in vitro and in vivo. HURP protein is highly charged with few predicted secondary and tertiary folding domains. Here we explore the effect of HURP on pure tubulin, and describe its ability to induce a new conformation of tubulin sheets that wrap around the ends of intact microtubules, thereby forming two concentric tubes. The inner tube is a normal microtubule, while the outer one is a sheet composed of tubulin protofilaments that wind around the inner tube with a 42.5 degrees inclination. We used cryo electron microscopy and unidirectional surface shadowing to elucidate the structure and conformation of HURP-induced tubulin sheets and their interaction with the inner microtubule. These studies clarified that HURP-induced sheets are composed of anti-parallel protofilaments exhibiting P2 symmetry. HURP is a unique MAP that not only stabilizes and bundles microtubules, but also polymerizes free tubulin into a new configuration. PMID- 17118404 TI - Serum levels of matrix metalloproteinase-9 and tissue inhibitors of metalloproteinases 1 in subacute sclerosing panencephalitis. AB - We determined the relationship between the serum concentrations of matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of metalloproteinases 1 (TIMP 1) in 33 patients with subacute sclerosing panencephalitis (SSPE) to investigate the function of the blood-brain-barrier (BBB) in SSPE. Serum MMP-9 and TIMP-1 levels were measured by ELISA. Serum MMP-9 levels and MMP-9/TIMP-1 ratios of SSPE patients in Papua New Guinea (n = 24), and those in Japan (n = 9) were significantly higher than the each control (MMP-9, p = 0.0390, and p = 0.0023, respectively; MMP-9/TIMP-1, p = 0.0319, and p = 0.0009, respectively). Serum MMP 9 levels and MMP-9/TIMP-1 ratios of SSPE patients with Jabbour stage III (n = 13) were significantly higher than those with Jabbour stage II (n = 18) (p = 0.003, and p = 0.0412, respectively). There were no significant differences of serum TIMP-1 levels between the SSPE patients and controls. High serum MMP-9 and MMP 9/TIMP-1 levels will promote brain invasion through the BBB by immunocompetent cells in the blood. Our findings suggest that the balance of serum MMP-9 and TIMP 1 levels modulate the inflammatory cascade of SSPE. PMID- 17118405 TI - Effects of carvedilol on transient outward and ultra-rapid delayed rectifier potassium currents in human atrial myocytes. AB - Carvedilol is a beta- and alpha(1)-adrenoceptor antagonist. It is widely used in the treatment of cardiovascular diseases including atrial arrhythmias. However, it is unclear whether carvedilol may affect the repolarization currents, transient outward K(+) current (I(to)) and ultra-rapid delayed rectifier K(+) current (I(Kur)) in the human atrium. The present study evaluated effects of carvedilol on I(to) and I(Kur) in isolated human atrial myocytes by whole-cell patch-clamp recording technique. We found that carvedilol reversibly inhibited I(to) and I(Kur) in a concentration-dependent manner. Carvedilol (0.3 microM) suppressed I(to) from 9.2+/-0.5 pA/pF to 4.8+/-0.5 pA/pF (P<0.01) and I(Kur) from 3.6+/-0.5 pA/pF to 1.9+/-0.3 pA/pF (P<0.01) at +50 mV. I(to) was inhibited in a voltage-dependent manner, being significantly attenuated at test potentials from +10 to +50 mV, whereas the inhibition of I(Kur) was independent. The concentration giving a 50% inhibition was 0.50 microM for I(to) and 0.39 microM for I(Kur). Voltage-dependence of activation, inactivation and time-dependent recovery from inactivation of I(to) were not altered by carvedilol. However, time to peak and time-dependent inactivation of I(to) were significantly accelerated, indicating an open channel blocking action. The findings indicate that carvedilol significantly inhibits the major repolarization K(+) currents I(to) and I(Kur) in human atrial myocytes. PMID- 17118406 TI - Folic acid supplementation delays atherosclerotic lesion development in apoE deficient mice. AB - Folic acid is a vitamin that when used as a dietary supplementation can improve endothelial function. To assess the effect of folic acid on the development of atherosclerosis, male apolipoprotein E-deficient mice fed a standard chow diet received either water (control group) or an aqueous solution of folic acid that provided a dose of 75 microg/kg/day, for ten weeks. At the time of sacrifice, blood was drawn and the heart removed. The study measured plasma homocysteine, lipids, lipoproteins, low-density lipoprotein (LDL) oxidation, isoprostane, paraoxonase, and apolipoproteins, and aortic atherosclerotic areas. In folic acid treated animals, total cholesterol, mainly carried in very low-density and low density lipoproteins, increased significantly, and homocysteine, HDL cholesterol, paraoxonase, and triglyceride levels did not change significantly. Plasma isoprostane and apolipoprotein (apo) B levels decreased. The resistance of LDL to oxidization and plasma apoA-I and apoA-IV levels increased with a concomitant decrease in the area of atherosclerotic lesions. The administration of folic acid decreased atherosclerotic lesions independently of plasma homocysteine and cholesterol levels, but was associated with plasma levels of apolipoproteins A-I, A-IV and B, and decreased oxidative stress. PMID- 17118407 TI - Parasite communities in Boops boops (L.) (Sparidae) after the Prestige oil-spill: detectable alterations. AB - Environmental pollution affects parasite populations and communities, both directly and through effects on intermediate and final hosts. In this work, we present a comparative study on the structure and composition of metazoan parasite communities in the bogue, Boops boops, from two localities (Galician coast, Spain) affected by the Prestige oil-spill (POS). We focus on the distribution of both individual parasite species and larger functional groupings by using both univariate and multivariate analyses. Our results indicate directional trends in community composition that might be related to the Prestige oil-spill disturbance of the natural coastal communities off Galicia. Endoparasite communities in B. boops reflected a notable change in the composition and abundance of the benthic fauna in the localities studied post-spill probably due to organic enrichment after the POS. PMID- 17118408 TI - An electrophysiological study on the interaction between emotional content and spatial frequency of visual stimuli. AB - Previous studies suggest that the magnocellular pathway, a visual processing system that rapidly provides low spatial frequency information to fast-responding structures such as the amygdala, is more involved in the processing of emotional facial expressions than the parvocellular pathway (which conveys all spatial frequencies). The present experiment explored the spatio-temporal characteristics of the spatial frequency modulation of affect-related neural processing, as well as its generalizability to non-facial stimuli. To that aim, the event-related potentials (ERPs) elicited by low-pass filtered (i.e., high spatial frequencies are eliminated) and intact non-facial emotional images were recorded from 31 participants using a 60-electrode array. The earliest significant effect of spatial frequency was observed at 135 ms from stimulus onset: N135 component of the ERPs. In line with previous studies, the origin of N135 was localized at secondary visual areas for low-pass filtered stimuli and at primary areas for intact stimuli. Importantly, this component showed an interaction between spatial frequency and emotional content: within low-pass filtered pictures, negative stimuli elicited the highest N135 amplitudes. By contrast, within intact stimuli, neutral pictures were those eliciting the highest amplitudes. These results suggest that high spatial frequencies are not essential for the initial affect related processing of visual stimuli, which would mainly rely on low spatial frequency visual information. According to present data, high spatial frequencies would come into play later on. PMID- 17118409 TI - Neural substrates of choice selection in adults and adolescents: development of the ventrolateral prefrontal and anterior cingulate cortices. AB - A heightened propensity for risk-taking and poor decision-making underlies the peak morbidity and mortality rates reported during adolescence. Delayed maturation of cortical structures during the adolescent years has been proposed as a possible explanation for this observation. Here, we test the hypothesis of adolescent delayed maturation by using fMRI during a monetary decision-making task that directly examines risk-taking behavior during choice selection. Orbitofrontal/ventrolateral prefrontal cortex (OFC/VLPFC) and dorsal anterior cingulate cortex (ACC) were examined selectively since both have been implicated in reward-related processes, cognitive control, and resolution of conflicting decisions. Group comparisons revealed greater activation in the OFC/VLPFC (BA 47) and dorsal ACC (BA 32) in adults than adolescents when making risky selections. Furthermore, reduced activity in these areas correlated with greater risk-taking performance in adolescents and in the combined group. Consistent with predictions, these results suggest that adolescents engage prefrontal regulatory structures to a lesser extent than adults when making risky economic choices. PMID- 17118410 TI - Inferring thought and action in motor neurone disease. AB - The traditional assumption that classical motor neurone disease (MND) invariably spares cognitive function is now recognised to be incorrect. Deficits have most commonly been demonstrated on executive tasks suggesting impaired function of frontal systems. Yet, crucial aspects of frontal lobe function have not hitherto been explored. The study used tests of theory of mind (ToM) (interpretation of cartoons and stories) to examine the ability of 16 patients with MND to interpret social situations and ascribe mental states to others. Only minor differences were elicited in the MND group as a whole compared to controls, and performance was not differentially affected for cartoons and stories requiring inference of another's mental state (mental) compared to control (physical) cartoons and stories. However, abnormalities were elicited on both mental and physical tasks in a subgroup of patients with bulbar signs. Moreover, examination of individual patient scores revealed a spectrum of performance ranging from normal to severely impaired. Errors were qualitatively similar to those seen in frontotemporal dementia (FTD). Performance on the ToM tasks was significantly correlated with conventional, untimed measures of executive function, suggesting that ToM deficits in MND are likely to be linked to a more general executive failure. The findings contribute to the understanding of ToM performance in neurodegenerative disease and provide further evidence of the association between MND and FTD. PMID- 17118411 TI - Bilateral damage to the sexually dimorphic medial preoptic area/anterior hypothalamus of male ferrets causes a female-typical preference for and a hypothalamic Fos response to male body odors. AB - Previous studies showed that bilateral lesions of the male ferret's preoptic area/anterior hypothalamus (POA/AH), centered in the sexually dimorphic nuclei present in this region, caused subjects to seek out a same-sex male, as opposed to a female conspecific. Male subjects with POA/AH lesions (which were also castrated and given estradiol) displayed female-typical receptive behavior in response to neck gripping by a stimulus male, implying that subjects' approaches to a same-sex conspecific were sexually motivated. We asked whether the effect of POA/AH lesions on males' partner preference reflects a shift in the central processing of body odorant cues so that males come to display a female-typical preference to approach male body odorants. Sexually experienced male ferrets in which electrolytic lesions of the POA/AH caused bilateral damage to the sexually dimorphic male nucleus (MN) resembled sham-operated females by preferring to approach body odors emitted from anesthetized male as opposed to female stimulus ferrets confined in the goal boxes of a Y-maze. This lesion-induced shift in odor preference was correlated with a significant increase in the ability of soiled male bedding to induce a Fos response in the medial POA of males with bilateral damage to the MN-POA/AH. No such partner preference or neural Fos responses were seen in sham-operated males or in other groups of males with POA/AH lesions that either caused unilateral damage or no damage to the MN-POA/AH. Male-typical hypothalamic processing of conspecifics' body odorants may determine males' normal preference to seek out odors emitted by female conspecifics, leading to mating and successful reproduction. PMID- 17118412 TI - Effect of delta9-tetrahydrocannabinol on quinine palatability and AM251 on sucrose and quinine palatability using the taste reactivity test. AB - Here we provide evidence that the cannabinoid agonist, Delta9 tetrahydrocannabinol (Delta9-THC) enhances quinine palatability and the CB1 antagonist/inverse agonist, AM251, reduces sucrose and quinine palatability using the taste reactivity test, which provides a direct measure of palatability independently of appetitive behavior. In Experiment 1, rats were treated with a low dose of Delta9-THC (0.5 mg/kg) or Vehicle 30 min, 60 min, 120 min or 240 min prior to a 5-min intraoral infusion of a highly unpalatable 0.05% quinine solution. Regardless of the post-injection interval, Delta9-THC reduced rejection of quinine. The Delta9-THC-induced palatability shift was reversed by AM251. In Experiment 2, rats were injected with either AM251 (1 mg/kg) or Vehicle prior to receiving a 5-min intraoral infusion of either 32% sucrose or 0.05% quinine solution. AM251 significantly decreased sucrose-elicited hedonic reactions across both time intervals; however, AM251 did not significantly modify the rejection of 0.05% quinine solution. When the concentration of the quinine solution was reduced to 0.01% in Experiment 3, AM251 enhanced quinine aversion. Although the range of concentrations of the solutions tested in the present data is limited, our results suggest that the cannabinoid system may modulate the palatability of ingested substances regardless of the palatability of the ingested substance. PMID- 17118413 TI - Saponins from Allium minutiflorum with antifungal activity. AB - Three saponins, named minutoside A (1), minutoside B (2), minutoside C (3), and two known sapogenins, alliogenin and neoagigenin, were isolated from the bulbs of Allium minutiflorum Regel. Elucidation of their structure was carried out by comprehensive spectroscopic analyses, including 2D NMR spectroscopy and mass spectrometry. The structures of the new compounds were identified as (25R)-furost 2alpha,3beta,6beta,22alpha,26-pentaol 3-O-[beta-D-xylopyranosyl-(1-->3)-O-beta-D glucopyranosyl-(1-->4)-O-beta-D-galactopyranosyl] 26-O-beta-D-glucopyranoside (1), (25S)-spirostan-2alpha,3beta,6beta-triol 3-O-beta-D-xylopyranosyl-(1-->3)-O beta-D-glucopyranosyl-(1-->4)-O-beta-D-galactopyranoside (2), and (25R)-furost 2alpha,3beta,5alpha,6beta,22alpha,26-esaol 3-O-[beta-D-xylopyranosyl-(1-->3)-O beta-D-glucopyranosyl-(1-->4)-O-beta-D-galactopyranosyl] 26-O-beta-D glucopyranoside (3). The isolated compounds were evaluated for their antimicrobial activity. All the novel saponins showed a significant antifungal activity depending on their concentration and with the following rank: minutoside B>minutoside C>>minutoside A. No appreciable antibacterial activity was recorded. The possible role of these saponins in plant-microbe interactions is discussed. PMID- 17118414 TI - Comparative pharmacokinetics of marbofloxacin in healthy and Mannheimia haemolytica infected calves. AB - The pharmacokinetics of marbofloxacin were investigated in healthy (n=8) and Mannheimia haemolytica naturally infected (n=8) Simmental ruminant calves following intravenous (i.v.) and intramuscular (i.m.) administration of 2 mg kg( 1) body weight. The concentration of marbofloxacin in plasma was measured using high performance liquid chromatography with ultraviolet detection. Following i.v. administration of the drug, the elimination half-life (t(1/2 beta)) and mean residence time (MRT) were significantly longer in diseased calves (8.2h; 11.13 h) than in healthy ones (4.6 h; 6.1 h), respectively. The value of total body clearance (CL(B)) was larger in healthy calves (3 ml min(-1) kg(-1)) than in diseased ones (1.3 ml min(-1) kg(-1)). After single intramuscular (i.m.) administration of the drug, the elimination half-life, mean residence time (MRT) and maximum plasma concentration (C(max)) were higher in diseased calves (8.0, 12 h, 2.32 microg ml(-1)) than in healthy ones (4.7, 7.4 h, 1.4 microg ml(-1)), respectively. The plasma concentrations and AUC following administration of the drug by both routes were significantly higher in diseased calves than in healthy ones. Protein binding of Marbofloxacin was not significantly different in healthy and diseased calves. The mean value for MIC of marbofloxacin for M. haemolytica was 0.1+/-0.06 microg ml(-1). The C(max)/MIC and AUC(24)/MIC ratios were significantly higher in diseased calves (13.0-64.4 and 125-618 h) than in healthy calves (8-38.33 and 66.34-328 h). The obtained results for surrogate markers of antimicrobial activity (C(max)/MIC, AUC/MIC and T > or = MIC) indicate the excellent pharmacodynamic characteristics of the drug in diseased calves with M. haemolytica, which can be expected to optimize the clinical efficacy and minimize the development of resistance. PMID- 17118415 TI - Synthesis and biological evaluation of some 17-picolyl and 17-picolinylidene androst-5-ene derivatives. AB - Starting from dehydroepiandrosterone (1) 17-picolyl (2), 17-picolinylidene (7), 17-picolinylidene-16-one (10 and 11), and 17-picolyl-16-one (15) derivatives of androst-5-ene were synthesized in one, two, four and five steps respectively. By the Oppenauer oxidation or dehydration of 2, 7, 10, and 11 with 2,3-dichloro-5,6 dicyano-1,4-benzoquinone (DDQ), the corresponding A and B ring modified derivatives 3, 5, 6, 8, 9, and 12-14 were obtained. The structure of 2 was unambiguously proved by the appropriate X-ray structural analysis. Compounds 3, 5, 9, 12-14 showed inhibitory activity against the enzyme aromatase. Antibacterial activity, toxicity to brine shrimp Artemia salina, antitumor activity against three tumor cell lines (human cervix carcinoma HeLa cells, human melanoma FemX cells, and human myelogenous leukemia K562 cells) and toxicity against peripheral blood mononuclear cells were evaluated. Three tested compounds, namely 11, 13, and 15, showed strong activity against all three cell lines, the IC(50) values being in the range of 4-10 microM. PMID- 17118416 TI - Insights into the carcinogenic mode of action of arsenic. AB - That arsenic can induce cancer in humans has been known since the late 17th century, yet how arsenic induces cancer has been the subject of numerous scientific publications. Various modes of action (MOA) have been proposed for arsenic's carcinogenicity. In this paper we review our previous studies on the ability of arsenicals to cause DNA damage, the relative inability of these arsenicals to induce point mutations, and the involvement of arsenicals in spindle disruption. We present new evidence that shows that reduced glutathione (GSH) can chemically reduce inactive pentavalent arsenicals to trivalent arsenicals which can disrupt tubulin polymerization, and show that reactive oxygen species (ROS) are most likely not involved in tubulin disruption. A hypothesis is also presented on how arsenic may induce stable chromosome aberrations (CAs) that can lead to cancer, thus supporting a role for genetic damage in the MOA for arsenic. We then propose promising areas of research that might give insight into the MOA of arsenic. PMID- 17118417 TI - Differential effects of five 'classical' scorpion beta-toxins on rNav1.2a and DmNav1 provide clues on species-selectivity. AB - In general, scorpion beta-toxins have been well examined. However, few in-depth studies have been devoted to species selectivity and affinity comparisons on the different voltage-activated Na(+) channels since they have become available as cloned channels that can be studied in heterologous expression systems. As a result, their classification is largely historical and dates from early in vivo experiments on mice and cockroach and fly larvae. In this study, we aimed to provide an updated overview of selectivity and affinity of scorpion beta-toxins towards voltage-activated Na(+) channels of vertebrates or invertebrates. As pharmacological tools, we used the classic beta-toxins AaHIT, Css II, Css IV, Css VI and Ts VII and tested them on the neuronal vertebrate voltage-activated Na(+) channel, rNa(v)1.2a. For comparison, its invertebrate counterpart, DmNav1, was also tested. Both these channels were expressed in Xenopus laevis oocytes and the currents measured with the two-electrode voltage-clamp technique. We supplemented this data with several binding displacement studies on rat brain synaptosomes. The results lead us to propose a general classification and a novel nomenclature of scorpion beta-toxins based on pharmacological activity. PMID- 17118418 TI - Development of a non-invasive biomonitoring approach to determine exposure to the organophosphorus insecticide chlorpyrifos in rat saliva. AB - Non-invasive biomonitoring approaches are being developed using reliable portable analytical systems to quantify dosimetry utilizing readily obtainable body fluids, such as saliva. In the current study, rats were given single oral gavage doses (1, 10, or 50 mg/kg) of the insecticide chlorpyrifos (CPF). Saliva and blood were then collected from groups of animals (4/time-point) at 3, 6, and 12 h post-dosing, and were analyzed for the CPF metabolite trichloropyridinol (TCP). Trichloropyridinol was detected in both blood and saliva at all doses and the TCP concentration in blood exceeded saliva, although the kinetics in blood and saliva were comparable. A physiologically based pharmacokinetic and pharmacodynamic (PBPK/PD) model for CPF incorporated a compartment model to describe the time course of TCP in blood and saliva. The model adequately simulated the experimental results over the dose ranges evaluated. A rapid and sensitive sequential injection (SI) electrochemical immunoassay was developed to monitor TCP, and the reported detection limit for TCP was 6 ng/L (in water). Computer model simulation in the range of the Allowable Daily Intake (ADI) or Reference Dose (RfD) for CPF (0.01-0.003 mg/kg/day) suggests that the electrochemical immunoassay has adequate sensitivity to detect and quantify TCP in saliva at these low exposure levels. However, to validate this approach, further studies are needed to more fully understand the pharmacokinetics of CPF and TCP excretion in saliva. These initial findings suggest that the utilization of saliva as a biomonitoring matrix, coupled to real-time quantitation and PBPK/PD modeling represents a novel approach with broad application for evaluating both occupational and environmental exposures to CPF. PMID- 17118419 TI - Discovery and characterization of the short kappaA-conotoxins: a novel subfamily of excitatory conotoxins. AB - We have characterized the defining members of a novel subfamily of excitatory conotoxins, the short kappaA-conotoxins (kappaA(S)-conotoxins). kappaA-conotoxins PIVE and PIVF (kappaA-PIVE and kappaA-PIVF) were purified from Conus purpurascens venom. Both peptides elicited excitatory activity upon injection into fish. kappaA-PIVE was synthesized for further characterization. The excitatory effects of kappaA-PIVE in vivo were dose dependent, causing hyperactivity at low doses and rapid immobilization at high doses, symptomatic of a type of excitotoxic shock. Consistent with these observations, kappaA-PIVE caused repetitive action potentials in frog motor axons in vitro. Similar results have been reported for other structurally distinct conotoxin families; such peptides appear to be required by most fish-hunting cone snails for the rapid immobilization of prey. Unexpected structure-function relationships were revealed between these peptides and two families of homologous conotoxins: the alphaA-conotoxins (muscle nAChR antagonists) and kappaA-conotoxins (excitotoxins), which all share a common arrangement of cysteine residues (CC-C-C-C-C). Biochemically, the kappaA(S) conotoxins more closely resemble the alphaA(S)-conotoxins than the other kappaA conotoxin subfamily, the long kappaA-conotoxins (kappaA(L)-conotoxins); however, kappaA(S)- and alphaA(S)-conotoxins produce different physiological effects. In contrast, the kappaA(S)-and kappaA(L)-conotoxins that diverge in several biochemical characteristics are clearly more similar in their physiological effects. PMID- 17118420 TI - Effect of exogenous interferon and an interferon inducer on western equine encephalitis virus disease in a hamster model. AB - Mice are used as models for western equine encephalitis virus (WEEV) infection, but high mortality is generally only seen with intracranial or intranasal challenge, while peripheral inoculation results in approximately 50% mortality and is not dose-dependent. Hamsters were therefore studied as a model for WEEV infection. Hamsters were highly sensitive to intraperitoneal (i.p.) infection with WEEV. Disease progression was rapid, and virus titers in serum, brain, liver, and kidney of infected hamsters peaked between 2 and 4 days post-virus inoculation (dpi). Foci of virus infection were detected in neurons of the cerebral cortex and midbrain. Pre-treatment i.p. with either interferon alfacon-1 (5 microg/kg/day) or with Ampligen (3.2 mg/kg/day) resulted in complete survival, reduced brain titers, and improved weight gain. This model of WEEV infection in hamsters appears to serve as a suitable model for the evaluation of potential therapeutic agents for the treatment of WEE disease. PMID- 17118421 TI - Infant color vision: prediction of infants' spontaneous color preferences. AB - Infants show spontaneous looking preferences among isoluminant chromatic stimuli [Adams, R. J. (1987). An evaluation of color preferences in early infancy. Infant Behavior and Development, 10, 143-150; Bornstein, M. H. (1975). Qualities of color vision in infancy. Journal of Experimental Social Psychology, 19 (3), 401 419.]. These differences in preference have often been called "hue" or "color" preferences, and attributed to differences in hue, but there are alternative explanations. Spontaneous preference variations remain after stimuli are equated for adult brightness, and thus cannot be attributed to adult-like brightness differences [Teller, D. Y., Civan, A., & Bronson-Castain, K. (2004). Infants' spontaneous color preferences are not due to adult-like brightness variations. Visual Neuroscience, 21 (3), 397-401]. In the present paper, we address three more alternative explanations: colorimetric purity; infant detection thresholds; and adult-like variations in saturation. Three experiments were conducted. In Experiment 1 we measured infants' spontaneous preferences for each of 22 different chromatic stimuli of varying dominant wavelength and colorimetric purity, each paired against the same white standard. In Experiment 2, we measured infants' chromatic detection thresholds. In Experiment 3, adult subjects made saturation matches between a blue-green standard and each of five other chromatic stimuli. Infant detection thresholds accounted for 34% of the variance in infant "hue" preferences, much more than colorimetric purity (2.4%) or adult saturation judgments (3%), but none of the three variables accounted for the majority of the variance. In our view, the most likely remaining option is that infants' spontaneous "hue" preferences indeed arise from preferences for the hues of stimuli that adults see as blue, purple and red. PMID- 17118422 TI - Human vergence eye movements initiated by competing disparities: evidence for a winner-take-all mechanism. AB - Vergence eye movements were elicited in human subjects at short latencies (approximately 70 ms) by applying binocular disparities briefly (200 ms) to large grating patterns (46 degrees wide, 35 degrees high). The positions of both eyes were recorded with the electromagnetic search coil technique. Using a dichoptic viewing arrangement (Wheatstone stereoscope), each eye viewed two overlapping 1-D sine waves that had the same orientation but different spatial frequencies. These two sine waves each had a binocular disparity that was 1/4 of its wavelength and the effect of varying their relative contrasts was examined (15 contrast ratios ranging from 0.125 to 8). The first experiment used horizontal gratings and recorded the vertical vergence responses when the two sine waves had spatial frequencies in the ratio 3:5 and vertical disparities of opposite sign. Initial vergence responses showed a highly nonlinear dependence on the contrast ratio. On average, when the contrast of one sine wave exceeded that of the other by a factor of >2.2, the sine wave with the higher contrast dominated responses and the sine wave with the lower contrast had almost no influence: winner-take-all. A second experiment, which used vertical gratings and recorded the horizontal vergence responses when the two sine waves had spatial frequencies in the ratio 3:5 and horizontal disparities of opposite sign, also uncovered nonlinear interactions but these were much more variable from one subject to another and, on average, one sine wave did not achieve complete dominance until its contrast exceeded that of the other by a factor of >4.5. When these two experiments were repeated with grating patterns in which the two sine waves had spatial frequencies in the ratio 3:7 and disparities of the same sign, similar nonlinear interactions were apparent. We attribute the nonlinear dependence on relative contrast to mutual inhibition between the neural elements processing the disparities of the two sine waves. We further suggest that this interaction will help to maintain binocular alignment on the objects in the plane of regard because the retinal images of those objects will tend to be better focused-and hence tend to have higher contrasts-than the images of objects in other depth planes. PMID- 17118423 TI - Transplacental transfer of phenyltins from a pregnant Dall's porpoise (Phocoenoides dalli) to her fetus. AB - The maternal to fetal transfer of monophenyltin (MPT), diphenyltin (DPT), and triphenyltin (TPT) was investigated for the first time in a cetacean species, a mother-fetus pair of Dall's porpoises (Phocoenoides dalli) collected off the Sanriku coast, Japan. The mother individual was contaminated by phenyltin compounds, and the muscle, blubber, and liver accounted for 90% of the total phenyltin burden. A two-generation phenyltin contamination via transplacental transfer was demonstrated. This study revealed that the transfer rate was 0.29% for total phenyltins to a fetus of about 6-month-old. Unlike in the mother, the fetal liver could not be considered the target organ for all the phenyltins studied. In a ca. 100kg-weight mother Dall's porpoise and its fetus of 2.21kg weight, approximately 153microg (TPT, 139microg; DPT, 9.4microg; MPT, 4.2microg) and 0.44microg of total phenyltins (TPT, 0.38microg; DPT, 0.03microg; MPT, 0.03microg) were found, respectively. The body burden of TPT consisted of 91% and 86% of the total phenyltin burdens in the maternal and fetal bodies, respectively. PMID- 17118424 TI - Fibroblast growth factors as regulators of stem cell self-renewal and aging. AB - Organ and tissue dysfunction which is readily observable during aging results from a loss of cellular homeostasis and reduced stem cell self-renewal. Over the past 10 years, studies have been aimed at delineating growth factors that will sustain and promote the self-renewal potential of stem cells and support the expansion of primitive stem cells in vitro and in vivo. Recently, strong evidence is emerging indicating that fibroblast growth factors (FGFs) play a crucial role in stem cell maintenance. FGFs belong to a family of polypeptide growth factors that are involved in multiple functions including cell proliferation, differentiation, survival and motility. In this review, we discuss the regulatory role of FGFs on hematopoietic stem cells (HSCs), neural stem cells (NSCs) and embryonic stem (ES) cells in maintaining stem cell self-renewal. These findings are useful and important to further our knowledge in stem cell biology and for therapeutic approaches. PMID- 17118425 TI - Inflammatory networks in ageing, age-related diseases and longevity. AB - Inflammation is considered a response set by the tissues in response to injury elicited by trauma or infection. It is a complex network of molecular and cellular interactions that facilitates a return to physiological homeostasis and tissue repair. The individual response against infection and trauma is also determined by gene variability. Ageing is accompanied by chronic low-grade inflammation state clearly showed by 2-4-fold increase in serum levels of inflammatory mediators. A wide range of factors has been claimed to contribute to this state; however, the most important role seems to be played by the chronic antigenic stress, which affects immune system thorough out life with a progressive activation of macrophages and related cells. This pro-inflammatory status, interacting with the genetic background, potentially triggers the onset of age-related inflammatory diseases as atherosclerosis. Thus, the analysis of polymorphisms of the genes that are key nodes of the natural immunity response might clarify the patho-physiology of age-related inflammatory diseases as atherosclerosis. On the other hand, centenarians are characterized by marked delay or escape from age-associated diseases that, on average, cause mortality at earlier ages. In addition, centenarian offspring have increased likelihood of surviving to 100 years and show a reduced prevalence of age-associated diseases, as cardiovascular disease (CVD) and less prevalence of cardiovascular risk factors. So, genes involved in CVD may play an opposite role in human longevity. Thus, the model of centenarians can be used to understand the role of these genes in successful and unsuccessful ageing. Accordingly, we report the results of several studies in which the frequencies of pro-inflammatory alleles were significantly higher in patients affected by infarction and lower in centenarians whereas age-related controls displayed intermediate values. These findings point to a strong relationship between the genetics of inflammation, successful ageing and the control of cardiovascular disease at least in men, in which these studies were performed. These data are also briefly discussed in the light of antagonistic pleiotropy theory and in order to pursuit a pharmacogenomics approach. PMID- 17118426 TI - A simple model system for age-dependent DNA damage and cancer. AB - Aging is the major risk factor for many human cancers. However, the mechanisms responsible for the effect of aging on tumor incidence are poorly understood, in part because few model systems are available to study age-dependent genomic instability. Furthermore, the role of DNA mutations in "normal aging" and "life span extension" is unclear. Our laboratory has developed a novel method to study aging in yeast based on the survival of non-dividing populations (chronological life span). Two major pathways have been identified that control chronological aging: the Ras/PKA/Msn2/4 and the Sch9 pathways. The downregulation of either of them promotes life span extension. Importantly, similar pathways (insulin/IGF-I like), regulate longevity in higher eukaryotes suggesting a common evolutionary origin for the life span-regulatory mechanisms. Moreover, both Ras and Sch9 are functional homologs of two major mammalian oncogenes (Ras and Akt), which underlines the close link between cancer and aging. By combining chronological life span with simple assays for the detection of DNA mutations and dedifferentiation we have developed a powerful system to identify genes that regulate genomic instability and understand the fundamental mechanisms that may be responsible for age-dependent DNA mutations and cancer in mammals. Here, we describe the use of this system to monitor the age-dependent accumulation of different types of DNA mutations including base substitutions, frame-shift mutations, and gross chromosomal rearrangements (GCRs). PMID- 17118427 TI - Volatile organic compounds in dwelling houses and stables of dairy and cattle farms in Northern Germany. AB - Farmers are exposed to a complex mixture of airborne substances which can represent a health hazard. Especially animal production on a farm can be a risk factor for respiratory diseases. Most studies in this context focused on bioaerosols, compounds attached to bioaerosols or on gases such as ammonia or hydrogen sulfide. Less attention was paid to volatile organic compounds (VOCs) which may also cause respiratory diseases. This pilot study presents results of measuring VOCs in the air of the dwellings and stables, as well as in the outdoor environment, of cattle farms in Northern Germany in spring and autumn. Farmers on all selected farms complained of symptoms such as asthma, rhinitis and conjunctivitis which occurred especially during work in the stable but also in the dwellings. The mean concentration of total VOC (TVOC) for the outdoor environment, the dwellings, and the stables were 100 microg m(-3), 763 microg m( 3), 322 microg m(-3) in spring and 143 microg m(-3), 544 microg m(-3), 595 microg m(-3) in autumn, respectively. There was no significant difference in season. TVOC concentrations in dwellings on farms were elevated when compared to dwellings in other environments, probably because of an additional exposure to fuels and chemicals used in agriculture. Aliphatic and alicyclic hydrocarbons, aromatic compounds, and terpenes were main compounds of the dwellings and ketones, alcohols, and esters of the stables. Terpene concentrations in the stables were low probably because straw, which was not a strong terpene emitter, was used instead of sawdust as floor covering. Large amounts of methylethylketone were encountered in the air of one stable, probably from animal exhalation caused by increased animal activity. However, it was unlikely that this caused respiratory symptoms. Generally it was likely that the concentrations of VOCs were too low to have health effects on their own. On the other hand, the VOC concentrations were in a multifactor concentration range in which health effects could occur depending on the interaction with other exposure factors. Fungal spores and bacteria were observed in addition to VOCs at the examined locations. Therefore it is possible that the observed VOCs played a role in exacerbating respiratory symptoms in this multifactor exposure. PMID- 17118428 TI - Transfer of lead from shot pellets to game meat during cooking. AB - We evaluated the transfer of Pb from shot to meat during the preparation of breasts of quails with 0, 1, 2 or 4 embedded Pb shot. A traditional Spanish recipe was used which utilizes vinegar during cooking to enhance the long-term preservation of the meat. The effect of the acidic conditions generated by the vinegar on Pb transfer was compared with the same recipe when the vinegar was substituted with water. The effect of up to 4 weeks of storage on Pb transfer has been also evaluated. The transfer of Pb from the embedded shot to the meat was much higher when cooking with vinegar than with water. However, the Pb transfer under acidic conditions did not increase significantly during long-term storage at room temperature. The consumption of only half a pickled quail per week with embedded shot may cause the provisional tolerable weekly intake (PTWI) of Pb by the Spanish consumer to be exceeded. PMID- 17118429 TI - Nitrogen and phosphorus dynamics of a re-wetted shallow-flooded peatland. AB - Minerotrophic peatlands play an important role in the regulation of water quality and quantity but due to drainage and agricultural land use most of these systems have lost this function. In Central Europe, many re-wetting projects have been implemented to restore wetlands for multifunctional purposes during the last years. The Pohnsdorfer Stauung (Northern Germany) is a eutrophic fen which was used for intensive agriculture for 40 years. The peatland is divided in two subareas by a small stream. In the scope of re-wetting measures, one subarea (westpolder) was flooded by blockading the main drainage ditch in 1996/97, a second subarea (eastpolder) was re-connected to the stream by an inlet and outlet in 2001. Nitrogen and phosphorus dynamics were investigated in the surface water of these systems over a 5-year (westpolder) and 3-year (eastpolder) period, and balances were calculated. In both polders high dynamics of nitrogen and phosphate concentrations were observed in the surface water. Nitrate peaked in the winter months, whereas ammonium, phosphate and organically bound nitrogen (N(org)) revealed highest values during summer. Daily balances for the eastpolder and annual balances for both polders were calculated. In both polders nitrate was retained but phosphate and N(org) were exported. Differences of the nutrient dynamics and the absolute and relative balances between the polders were due to the different hydrology: the eastpolder received a high nitrate load by the stream, in the westpolder nutrient loads are low and internal nutrient cycles are dominating. During the summer months, high ammonium concentrations, oxygen depletion, and phosphate release occur in the warm, stagnant water of both polders. Normally, high phosphate and ammonium concentrations did not coincided with discharge phases from the polders. However, in August 2002 after heavy rainfall high phosphorus and ammonium discharge was observed. One important factor influencing the detected nutrient dynamics and balances is the change from a drained and fertilized peatland to a flooded wetland in only a few years. We conclude that continuous water flow and stable water levels are desired for flooded peatlands to prevent high concentrations of ammonium and phosphate and to utilize the potential for nitrate retention. In degenerated peatlands, flooding should only be regarded as an easy technological solution in order to conserve the peat mineralization, and to form the foundation for the further development of the peatland to a more efficient nutrient retaining system. PMID- 17118430 TI - The effect of pre-aeration on the purification processes in the long-term performance of a horizontal subsurface flow constructed wetland. AB - Different conditions (water level, oxygen supply) prevailing in both beds of the Kodijarve double-bed horizontal subsurface flow (HSSF) constructed wetland (CW) (Southern Estonia; constructed in 1996, total area 312.5 m(2), 40 pe) provide the opportunity to compare how different operational methods have altered the efficiency of the purification processes inside the HSSF CW. In summer 2002 a vertical subsurface flow (VSSF) CW (total area 37.4 m(2)) was added as the first stage of the system. Data from 18 sampling wells installed in Kodijarve HSSF CW from two periods is compared: 1st period -- January 2000-April 2002 (before the VSSF CW was built); 2nd period --October 2002-December 2004 (after the construction of the VSSF filter). The VSSF CW has remarkably improved aerobic conditions in both beds of the HSSF. Apart from total phosphorus concentrations in the right bed and nitrate nitrogen concentrations in the outflow of both beds, all of the water quality indicators (dissolved oxygen, total suspended solids, biological oxygen demand, ammonia nitrogen, nitrite nitrogen, total nitrogen and total iron) improved after the construction of the VSSF filter. Typically, purification processes in the HSSF CW were dependent on oxygen supply, which was partly influenced by the water level inside the filter beds. PMID- 17118431 TI - The interval between prothrombin time tests and the quality of oral anticoagulants treatment in patients with chronic atrial fibrillation. AB - BACKGROUND: The incidence of stroke in patients with atrial fibrillation (AF) can be significantly reduced with warfarin therapy especially if optimally controlled. OBJECTIVES: To evaluate the effect of the interval between consecutive prothrombin time measurements on the time in therapeutic range (INR 2 3) in a cohort of patients with AF on chronic warfarin treatment in the community. METHODS: All INR measurements available from a relatively large cohort of patients with chronic AF were reviewed and the mean interval between consecutive INR tests of each patient was correlated with the time in therapeutic range (TTR). RESULTS: Altogether 251,916 INR measurements performed in 4408 patients over a period of seven years were reviewed. Sixty percent of patients had their INR measured on average every 2 to 3 weeks and most others were followed at intervals of 4 weeks or longer. A small proportion (3.6%) had their INR measured on average every week. A significant decline in the time in therapeutic range was observed as the intervals between tests increased. At one to three weeks interval the TTR was 48%, at 4 weeks interval 45% and at 5 weeks 41% (P<0.0005). A five percent increment in TTR was observed if more tests were performed at multiplications of exactly 7 days (43% vs 48% P<0.0001). CONCLUSIONS: A better control with an increase in the TTR was observed in patients with atrial fibrillation if prothrombin time tests are performed at regular intervals of no longer than 3 weeks. PMID- 17118432 TI - Monitoring Unfractionated Heparin (UFH) therapy: which Anti-Factor Xa assay is appropriate? AB - INTRODUCTION: Anti-Factor Xa (Anti-Xa) assays specifically determine the anticoagulant activity of UFH by measuring the ability of heparin-bound Antithrombin (AT) to inhibit a single enzyme, Factor Xa (FXa). Recent improvements in the automation, cost-effectiveness and accessibility of the assay to clinicians, have resulted in the Anti-Xa assay becoming a part of daily clinical practice in many institutions. OBJECTIVES: We hypothesized that different Anti-Xa assays have different applicability for use in clinical settings, depending on the amount of UFH administered. This was investigated in a tertiary paediatric institution. MATERIALS AND METHODS: Samples were collected from children receiving Low-dose of UFH of at least 10 IU/kg/h, with or without a previous bolus of up to 25 IU/kg/h, within the previous 6 h in the PICU and HDU. High-dose UFH population consisted of children undergoing Cardiac Catheterization (CC), who received a bolus of UFH of 100 IU/kg body weight, 30 min prior to sampling. RESULTS AND CONCLUSIONS: The Anti-Xa activity for a given dose of UFH was found to vary significantly based on the Anti-Xa assay and the population being monitored. Our study suggests that the MODIFIED COMATIC Anti-Xa assay provides the best physiological measure of the UFH effect in children, as it does not introduce sources of error, such as exogenous AT, which may increase the measured ant Factor Xa activity, nor Dextran Sulphate which can displace plasma protein bound heparin and once again leading to falsely elevated assay results. Further studies that include assessment of clinical outcomes are required to confirm the applicability of use of this particular assay in monitoring UFH therapy. PMID- 17118433 TI - Mammalian embryo co-culture: trials and tribulations of a misunderstood method. AB - Embryo-somatic cell co-culture was devised over 40 years ago in an attempt to improve the development and viability of mammalian preimplantation embryos generated and cultured in vitro. While initial endeavours were successful in this respect, other studies soon highlighted a number of significant long-term detrimental impacts of this approach. Surprisingly little is known about the mechanisms underlying the beneficial effects of co-culture, although the production of embryotrophic compounds, modulation of nutrient profile, protection against culture-induced stress and/or toxin clearance are all contenders. The extent to which the inadvertent exposure of embryos to serum accounts for many of these effects remains open to question. Although the popularity of somatic cell co-culture has recently declined in favour of the use of sequential media due to concerns associated with its risk of disease transmission and long-term sequelae, we argue that complete dismissal of this technique is ill advised, given that our limited understanding of basic somatic cell interactions has prevented us from fully exploiting its potential. In this respect, there is some merit in focussing future research strategies based on reconstructed maternal tract tissue. Although the use of co-culture in clinical practice is unacceptable and its implementation in domestic species for commercial purposes should be viewed with diffidence, this technique can still provide a wealth of information on the development of novel, more physiological embryo in vitro culture systems. The proviso for acquiring such information is to gain a fuller understanding of the culture requirements/biochemistry of somatic cells and their interaction with the early conceptus. PMID- 17118434 TI - Factors affecting the fertility of high producing dairy herds in northeastern Spain. AB - Infertility has been often correlated to a rising milk yield in high producing dairy cattle. The aim of the present study was to evaluate, using logistic regression procedures, the effects of several management indicators on the fertility of four dairy herds in northeastern Spain. Data derived from 10,965 artificial insemination (AI). The factors examined were: herd, milking frequency (three versus two milkings per day), lactation number, previous twinning and disorders such as placenta retention and pyometra, milk production at AI, the inseminating bull, season (warm versus cool period) and year effects, AI technician and repeat breeding syndrome (cows undergoing four or more AI). Our findings indicated no effects on fertility of the herd, year of AI, previous twining, placenta retention and pyometra and milk production at AI. Based on the odds ratios, the likelihood of pregnancy decreased: in cows milked three times per day (by a factor of 0.62); for each one unit increase in lactation number (by a factor of 0.92); for inseminations performed during the warm period (by a factor of 0.67); in repeat breeder cows (by a factor of 0.73); and when 3 of the 45 inseminating bulls included in the study were used (by factors of 0.35, 0.43 and 0.44, respectively). Of the 13 AI technicians participating in the study, 3 were related to a fertility rate improved by odds ratios of 1.86, 1.84 and 1.30, respectively, whereas 2 technicians gave rise to fertility rates reduced by odds ratios of 0.64 and 0.49, respectively. Under our study conditions, management practices were able to compensate for the effects of previous twining and reproductive disorders such as placenta retention and pyometra. However, fertility was significantly affected by the factors milking frequency, AI technician, inseminating bull, repeat breeding syndrome, lactation number and AI season. PMID- 17118435 TI - Characterization of rainbow trout egg quality: a case study using four different breeding protocols, with emphasis on the incidence of embryonic malformations. AB - The aim of this study was to set up a methodology to accurately evaluate the effects of various husbandry practices on a fish broodstock based on the developmental potential of the egg. For that purpose, long-short photoperiod manipulations (tested twice, PM1 and PM2 groups), spawning induction by injection of a GnRH analog (SI group), and a 16-day post-ovulatory ageing of eggs (POA group) were used in rainbow trout (Oncorhynchus mykiss). Females without any treatment were used as a control group. Survival at eying (E) and yolk-sac resorption (YSR) were recorded and malformations at YSR were monitored according to a detailed typology that included cyclopia, torsion, incomplete YSR, prognathia, and others. Egg weight was also monitored. A deleterious effect of photoperiod manipulation was observed on egg quality in both PM1 and PM2 groups. Incomplete YSR appeared as the predominant malformation while cyclopia type was nearly absent. In the SI group, a limited effect on egg quality was observed in comparison to the other experimental groups, although the percentage of normal alevins at YSR was significantly lower than in the control group. Finally, the most important effects on egg quality were observed in the POA group. The percentage of normal alevins was only 14+/-6% (mean+/-95% confidence interval) while the percentage of malformed embryos reached 49+/-11%. The proportion of cyclopia was significantly higher than in the control group. In conclusion, the type of egg quality alteration is extremely dependent on the applied breeding protocols, and the proposed methodology is able to discriminate those experimental conditions even when the impact on egg quality is limited. PMID- 17118436 TI - Influence of sire and sire breed (Gyr versus Holstein) on establishment of pregnancy and embryonic loss in lactating Holstein cows during summer heat stress. AB - Heat stress has negative effects on pregnancy rates of lactating dairy cattle. There are genetic differences in tolerance to heat stress; Bos taurus indicus (B. t. indicus) cattle and embryos are more thermotolerant than Bos taurus taurus (B. t. taurus). In the present study, the effects of sire and sire breed on conception and embryonic/fetal loss rates of lactating Holstein cows during the Brazilian summer were determined. In Experiment 1, cows (n=302) were AI after estrus detection or at a fixed-time with semen from one Gyr (B. t. indicus) or one Holstein sire (B. t. taurus). Pregnancy was diagnosed 80 days after AI. In Experiment 2, cows (n=811) were AI with semen from three Gyr and two Holstein sires. Pregnancy was diagnosed at 30-40 and at 60-80 days after AI. Cows diagnosed pregnant at the first examination but non-pregnant at the second were considered as having lost their embryo or fetus. Data were analyzed by logistic regression. The model considered the effect of sire within breed, sire breed, days postpartum, period of lactation, and AI type (AI after estrus versus fixed time). There was no effect of the AI type, days postpartum or milk production on conception or embryonic loss rates. The use of Gyr bulls increased pregnancy rate when compared to Holstein bulls [9.1% (60/657) versus 5.0% (23/456), respectively, P=0.008; data from Experiments 1 and 2 combined]. Additionally, in Experiment 2, cows inseminated using semen from sire #4 (Gyr) had lower embryonic loss (10%) when compared with other B. t. indicus (35.3% and 40%) or B. t. taurus sires (18.2% and 38.5%, P=0.03). In conclusion, the use of B. t. indicus sires may result in higher conception rates in lactating Holstein cows during summer heat stress. Moreover, sire can affect embryonic loss and selection of bulls according to this criterion may result in higher parturition rates in lactating Holstein cows. PMID- 17118437 TI - Effect of serum starvation on the efficiency of nuclear transfer using odd-eyed white cat fibroblasts. AB - In the present study, we compared in vitro and in vivo development of nuclear transfer (NT) embryos derived from serum-starved or non-serum-starved odd-eyed cat skin fibroblast cells. Flow cytometry analyses revealed that a higher percentage of cells were in the G0/G1 phase after serum starvation (89.3%) as compared with non-serum-starved cells (73.8%, P<0.05). Frequency of cleavage and development to the blastocyst stage was not different between the serum-starved or non-serum-starved treatment group, 67.9 and 12.5% versus 73.0 and 10.2%, respectively (P>0.05). After transfer of two to four-cell NT embryos derived from starved and non-starved fibroblasts, three of nine (33%) and one of nine (11%) recipients delivered three live male (plus, one dead) and two live male kittens, respectively. Of the five live-cloned kittens, one died from diarrhea at 3 weeks of age and the other four kittens are growing at a normal rate. The cloned kittens are blue-eyed and have functional auditory systems, including clones of the odd-eyed deaf Turkish Angora cat. Subsequent DNA analysis of nine-cat specific microsatellite loci confirmed that all of the cloned kittens were identical to the odd-eyed donor male, but a point mutation occurred in the dead fetus at the FCA 290 marker. PMID- 17118438 TI - Aging studies of light cured dimethacrylate-based dental resins and a resin composite in water or ethanol/water. AB - OBJECTIVE: To study the aging of neat resins, prepared from bis-GMA, UDMA, D(3)MA or a mixture of bis-GMA/UDMA/D(3)MA (65/20/15 w/w/w), in water or 75% (v/v) ethanol/water 37 degrees C. Also the study of aging of Heliomolar RO, which is a radiopaque, microfilled, light cured composite, the resin matrix of which is the copolymer of bis-GMA/UDMA/D(3)MA (65/20/15 w/w/w). METHODS: Samples of neat resins and Heliomolar RO prepared by light curing were immersed in water or 75% (v/v) ethanol/water 37 degrees C, for 1, 7 or 30 days. Then the flexural and tensile strength were determined. The fractured surface of samples after the flexural tests was observed by scanning electron microscopy. RESULTS: Bis-GMA and copolymer resin did not showed any significant change in mechanical properties after immersion in water or 75% (v/v) ethanol/water 37 degrees C, for 30 days. On the contrary UDMA, D(3)MA and the composite Heliomolar RO showed a significant decrease. SIGNIFICANCE: The results obtained showed that the effect of aging in water or ethanol/water solution on mechanical properties of a light cured dimethacrylate resin depends on the chemical structure of resin. In the case of resin composite this effect depends on the filler-matrix bond strength. PMID- 17118439 TI - Mechanical and physical properties of carbon-graphite fiber-reinforced polymers intended for implant suprastructures. AB - OBJECTIVES: Mechanical properties and quality of fiber/matrix adhesion of poly(methyl methacrylate) (PMMA)-based materials, reinforced with carbon-graphite (CG) fibers that are able to remain in a plastic state until polymerization, were examined. METHODS: Tubes of cleaned braided CG fibers were treated with a sizing resin. Two resin mixtures, resin A and resin B, stable in the fluid state and containing different cross-linking agents, were reinforced with CG fiber loadings of 24, 36, and 47 wt% (20, 29, and 38 vol.%). In addition, resin B was reinforced with 58 wt% (47 vol.%). After heat-polymerization, flexural strength and modulus were evaluated, both dry and after water storage. Coefficient of thermal expansion, longitudinally and in the transverse direction of the specimens, was determined. Adhesion between fibers and matrix was evaluated with scanning electron microscopy (SEM). RESULTS: Flexural properties and linear coefficient of thermal expansion were similar for both fiber composites. With increased fiber loading, flexural properties increased. For 47 wt% fibers in polymer A the flexural strength was 547.7 (28.12) MPa and for polymer B 563.3 (89.24) MPa when water saturated. Linear coefficient of thermal expansion was for 47 wt% CG fiber reinforced polymers; -2.5 x 10(-6) degrees C-1 longitudinally and 62.4 x 10(-6) degrees C-1 in the transverse direction of the specimens. SEM revealed good adhesion between fibers and matrix. More porosity was observed with fiber loading of 58 wt%. CONCLUSIONS: The fiber treatment and the developed resin matrices resulted in good adhesion between CG fibers and matrix. The properties observed indicate a potential for implant-retained prostheses. PMID- 17118440 TI - Effect of thermal cycling on composites reinforced with two differently sized silica-glass fibers. AB - OBJECTIVES: To evaluate the effects of thermal cycling on the flexural properties of composites reinforced with two differently sized fibers. METHODS: Acid-washed, woven, fused silica-glass fibers, were heat-treated at 500 degrees C, silanized and sized with one of two sizing resins (linear poly(butyl methacrylate)) (PBMA), cross-linked poly(methyl methacrylate) (PMMA). Subsequently the fibers were incorporated into a polymer matrix. Two test groups with fibers and one control group without fibers were prepared. The flexural properties of the composite reinforced with linear PBMA-sized fibers were evaluated by 3-point bend testing before thermal cycling. The specimens from all three groups were thermally cycled in water (12,000 cycles, 5/55 degrees C, dwell time 30 s), and afterwards tested by 3-point bending. SEM micrographs were taken of the fibers and of the fractured fiber reinforced composites (FRC). RESULTS: The reduction of ultimate flexural strength after thermal cycling was less than 20% of that prior to thermal cycling for composites reinforced with linear PBMA-sized silica-glass fibers. The flexural strength of the composite reinforced with cross-linked PMMA-sized fibers was reduced to less than half of the initial value. SIGNIFICANCE: This study demonstrated that thermal cycling differently influences the flexural properties of composites reinforced with different sized silica-glass fibers. The interfacial linear PBMA-sizing polymer acts as a stress-bearing component for the high interfacial stresses during thermal cycling due to the flexible structure of the linear PBMA above Tg. The cross-linked PMMA-sizing, however, acts as a rigid component and therefore causes adhesive fracture between the fibers and matrix after the fatigue process of thermal cycling and flexural fracture. PMID- 17118441 TI - Brain acetylcholinesterase as a marine pesticide biomarker using Brazilian fishes. AB - Brain acetylcholinesterase (AChE) of some fishes from the coast of Rio de Janeiro State was studied as a possible pesticide biomarker in marine environmental monitoring. AChE specific activity in brain varied from 145 to 530 U/g of proteins and the Michaelis-Menten constant (K(M)) for acetylthiocholine varied from 104 to 291 microM among the 20 species studied. The enzyme sensitivity to methyl paraoxon, evaluated by the inhibition kinetic constants, shows that some species (Paralonchurus brasiliensis and Genidens genidens) are more sensitive (IC50-30 min=455 and 468 nM, respectively). The less sensitive Merluccius hubbsi and Percophis brasiliensis (IC50-30 min=3339 and 3259 nM, respectively) belong to the super-order Paracanthopterygii, which includes the more ancient species. On the other hand, more susceptible species belong to the super-order Acanthopterygii, which includes more recent species. These results suggest a possible evolutionary linkage for AChE sensitivity to methyl paraoxon. The application of inhibition kinetic constants for fish brain AChE in phylogenetic studies is still being investigated. The results have shown that a fish sentinel species should have the highest brain AChE level among the more sensitive ones. PMID- 17118442 TI - The preservation of phenotype and functionality of dendritic cells upon phagocytosis of polyelectrolyte-coated PLGA microparticles. AB - Biodegradable microparticles (MP) represent a promising and efficient delivery system for parenteral vaccination. Recently, MP have also been explored as tool for the ex vivo antigen loading of professional antigen-presenting cells such as dendritic cells (DC) to be used as cellular vaccines. The purpose of this study was to investigate various polycationic coatings on poly(lactide-co-glycolide) (PLGA) MP, with regard to their effect on phenotypic and functional maturation of monocyte-derived DC (MoDC) that had previously been loaded with the MP in vitro. The preparation and concomitant coating of the PLGA was performed by means of a solvent extraction/evaporation method using a recently developed microextrusion based technique. The polyelectrolytes tested for MP coating encompassed aminodextran, chitosan, poly(ethylene imine) (PEI), poly(L-lysine) and protamine. Uncoated and differently coated PLGA MP were fed to immature MoDC, which ingested efficiently the different MP types irrespective of their surface coating. The MP loaded immature MoDC were then matured with the help of a cytokine/PGE-2 maturation cocktail. Here, the presence of the ingested MP did not affect the MoDC maturation in terms of expression of the surface markers CD80, CD83, CD86, HLA-DR and MMR, irrespective of the MP surface coating. Importantly, none of the PLGA MP types alone induced significant maturation of MoDC in the absence of the maturation cocktail. MP-loaded and subsequently matured MoDC expressed high levels of the chemokine receptor CCR7, whose functional activity was evidenced by the migration of MoDC towards CCL21, irrespective of the presence of ingested MP. Further, MP-loaded and subsequently matured MoDC also secreted comparable amounts of IL-10 and IL-12p70, irrespective of the presence of ingested MP except for PEI coated PLGA MP, which enhanced significantly the secretion of IL-12p70 in mature MoDC. In conclusion, phenotypic and functional maturation of MoDC by means of a maturation cocktail remained unchanged irrespective of the presence of previously ingested differently coated PLGA MP. This offers interesting perspectives for using these particulate systems together with entrapped antigens for ex vivo loading of MoDC in view of cellular immunotherapy. PMID- 17118443 TI - Superporous hydrogels containing poly(acrylic acid-co-acrylamide)/O-carboxymethyl chitosan interpenetrating polymer networks. AB - Superporous hydrogels containing poly(acrylic acid-co-acrylamide)/O-carboxymethyl chitosan interpenetrating polymer networks (SPH-IPNs) were prepared by cross linking O-carboxymethyl chitosan (O-CMC) with glutaraldehyde (GA) after superporous hydrogel (SPH) was synthesized. The structures of the SPH-IPNs were characterized with FT-IR, 13C-NMR and DSC. SEM, CLSM and light images revealed that the SPH-IPNs possessed both the IPN network and large numbers of pores and the cross-linked O-CMC molecules were located on the peripheries of these pores. The swelling behavior of SPH-IPNs was dependent on the O-CMC content, GA amount and cross-linking time. Due to the cross-linked O-CMC network, in vitro muco adhesive force and mechanical properties, including compression and tensile modulus, of the SPH-IPN were greatly improved when compared with the CSPH. An enhanced loading capacity for insulin could be obtained by the SPH-IPNs as compared to non-porous hydrogel and CSPH, and more than 90% of the insulin was released within 1 h. With the improved mechanical properties, in vitro muco adhesive force and loading capacities, the SPH-IPN may be used as a potential muco-adhesive system for peroral delivery of peptide and protein drugs. PMID- 17118444 TI - Improved osteogenic differentiation of human marrow stromal cells cultured on ion induced chemically structured poly-epsilon-caprolactone. AB - The ability to control cell proliferation/differentiation, using material surface, is a main goal in tissue engineering. The objective of this study was to evaluate the attachment, proliferation and differentiation to the osteoblastic phenotype of human marrow stromal cells (MSC) when seeded on poly-epsilon caprolactone (PCL) thin films before and after irradiation with 10 keV He+. The polymeric surface was characterized as surface chemical structure and composition, roughness and morphology on the micro- and nano-scale, wettability and surface free energy parameters. MSC were obtained from patients undergoing routine hip replacement surgery, expanded in vitro and cultured on untreated PCL and He+ irradiated PCL films for up to 4-5 weeks in osteogenic medium. He+ irradiation led to slight smoothening of the surface and different nanoscale surface chemical structure, while surface free energy resulted unchanged in comparison to untreated PCL. The results from biological testing demonstrated that early attachment and further proliferation, as well as osteoblastic markers, were higher for MSC on He+-irradiated PCL. In conclusion, the change of PCL surface properties induced by ion beam irradiation is confirmed to enhance the adhesion of MSC and support their differentiation. PMID- 17118445 TI - Expression and functional activity of ryanodine receptors (RyRs) during skeletal muscle development. AB - Two isoforms of ryanodine receptors are expressed in skeletal muscles, RyR1 and RyR3. We investigated the relative level of expression of RyRs in developing murine skeletal muscles using [3H]ryanodine binding and immunoprecipitation experiments. In the diaphragm RyR3 accounted for 11% of total RyRs in 5-day-old mice and for 3% of total RyRs in 60-day-old mice. In hindlimb muscles, RyR3 accounted for 3% and 1% of total RyRs in 5-day-old and adult mice, respectively. The activity of RyR1 channels in native microsomal vesicles from murine muscles was found to be as low as 35% of that measured after CHAPS exposure, while no inhibition was observed for RyR3. CHAPS sensitivity of recombinant RyR1 and RyR3 expressed in HEK293 cells was also investigated. The activity of recombinant RyR1 but not RyR3 channels was found to be inhibited in native conditions, suggesting that this property may not be dependent on a muscle environment. PMID- 17118446 TI - Folate levels and methylation of CDKI proteins. PMID- 17118447 TI - The GST deletions and NQO1*2 polymorphism confers interindividual variability of response to treatment in patients with acute myeloid leukemia. AB - Functional polymorphisms in the genes encoding detoxification enzymes could modify the response to treatment in acute myeloid leukemia and therefore affect the final clinical outcome. In the present study, we genotyped 153 patients diagnosed with de novo acute myeloid leukemia (AML) to clarify the influence of the genetic polymorphisms CYP1A1*2A, CYP3A4*1B, CYP2E1*5B, del{GSTT1}, del{GSTM1}, and NQO1*2 on disease outcome. The del{GSTM1} showed a higher frequency in females (62%) than in males (41%) (P=0.01). The number of functional NQO1 alleles influenced the response to induction therapy; 81% (55/68) NQO1 negative patients, 69% (28/41) heterozygous patients, and 27% (2/7) homozygous patients achieved complete remission (CR) (P=0.04). The presence of GST deletions was associated with a lower probability of disease-free survival (DFS) and this effect was more relevant in male patients. Males with del{GSTM1} showed a 28% DFS versus 57% DFS for undeleted GSTM1 (P=0.04). Similarly, males with undeleted GSTM1 and GSTT1 showed a 64% DFS versus 34% DFS for males with at least one GST deletion (P=0.05). This study suggests that the NQO1*2 polymorphism is relevant to the patient's response to induction therapy and that GST deletions influence treatment outcome after chemotherapy, especially in male patients. PMID- 17118448 TI - Kid's stuff: the nature and impact of peer and sibling violence on younger and older children. AB - OBJECTIVE: It is widely presumed that when children are hit by peers or siblings, it is not as serious as similar acts between adults or older youth, which would be termed, "assaults" and "violent crimes". The goal of this study was to compare the violent peer and sibling episodes of younger children to those of older youth in terms of their seriousness and association with symptoms that might indicate traumatic effects. METHOD: The study collected reports of past year's violent victimizations and childhood symptoms in a national probability telephone sample of 2030 children and youth ages 2-17. The experiences of 10-17-year olds were obtained via self-reports and those of the 2-9-year olds from caregivers. RESULTS: The younger children's peer and sibling victimizations were not less serious than the older youth on the dimensions of injury, being hit with an object that could cause injury or being victimized on multiple occasions. Younger children and older youth also had similar trauma symptom levels associated with both peer and sibling victimization. CONCLUSION: There was no basis in this study for presuming peer and sibling victimizations to be more benign when they involve younger children. The findings provide justification for being concerned about such peer and sibling violence in schools and families and for counting such victimizations in victimization inventories and clinical assessments. PMID- 17118449 TI - Internalization and molecular interactions of human CD21 receptor. AB - The human CD21 is a receptor for cleavage fragments of the third complement component and for Epstein-Barr virus. Previous mutational studies showed that the cytoplasmic domain of CD21 is absolutely required for internalization of either ligand. With the exception of CD19, CD81, Leu-13 and CD35 that can form a complex with CD21 at the cell surface, no other partner that interacts with the hCD21 transmembrane or the cytoplasmic domain was identified. We investigated the internalization capacity of hCD21 tail mutants in the absence of B cell receptor cross-linking by using stable murine B cell transfectants. We provide evidence that at least two internalization motifs are activated when hCD21 binds a monoclonal antibody. In order to identify the cellular proteins that interact with the hCD21 transmembrane and cytoplasmic domains, we combined a mutational mapping with a two-hybrid system approach both in yeast and in mammalian cells. We identified four novel partners that are involved in intracellular trafficking, sorting or cytoskeleton remodeling and we mapped the hCD21 transmembrane and tail subdomains they interact with. We discuss the potential physiological significance of these findings in the context of hCD21 internalization and intracellular trafficking. PMID- 17118450 TI - Methotrexate regulates the expression of glucocorticoid receptor alpha and beta isoforms in normal human peripheral mononuclear cells and human lymphocyte cell lines in vitro. AB - MTX is an effective therapy for autoimmune-inflammatory diseases. The mechanisms that mediate these actions are not completely clear. It is accepted that many of these effects are mediated through the release of adenosine with the activation of the adenosine receptor A2. MTX is used as a steroid sparing agent. An improved in vitro GC cell sensitivity in GC insensitive asthma patients has been demonstrated after MTX treatment. Most GC actions are mediated by the GCR. The effect of MTX on GCRs expression has not been previously evaluated. Therefore, we evaluate if MTX regulates the expression of glucocorticoid receptors, increasing the expression of the active receptor (GCR alpha) and/or decreasing the expression of the dominant negative receptor (GCR beta). We show that MTX increases the mRNA and protein levels of GCR alpha and decreases or leaves unchanged the protein expression of the GCR beta in CEM cells in culture. This effect was also observed in other lymphocytes (Jurkat and Raji) and in PBMNC from healthy volunteers. We also show that upon MTX treatment PBMC from normal volunteers exhibit a higher sensitivity to DEX inhibition on LPS-induced TNF alpha release. To explore if these actions are mediated by adenosine through the adenosine receptor A2 we evaluate the effect of adenosine on the GCRs expression and the effect of an A2 receptor blocker (DMPX) on MTX effects on GCRs expression. Our results show that adenosine does not mimic and DMPX can enhance MTX effects on these receptors. We conclude that MTX increases the GCR alpha/GCR beta ratio of expression in lymphocytes which could mediate its previously reported effects in improving cell glucocorticoid sensitivity. These actions are not mediated by the adenosine receptor A2. PMID- 17118451 TI - Porcine T-cell receptor beta-chain: a genomic sequence covering Dbeta1.1 to Cbeta2 gene segments and the diversity of cDNA expressed in piglets including novel alternative splicing products. AB - Porcine TCRbeta-chain cDNA clones were isolated from thymic and peripheral blood lymphocytes of piglets. Using these nucleotide sequences, a genomic 18kbp sequence stretch covering Dbeta1 to Cbeta2 gene segments was identified, which revealed that the porcine TCRbeta-chain locus consists of two sets of Dbeta-Jbeta Cbeta gene groups with each set having a Dbeta gene segment, seven Jbeta gene segments and a down stream Cbeta gene segment composed of four exons. This structure is consistent with other known mammalian TCRbeta-chain loci. With this genomic information, TCRbeta-chain clones from cDNA libraries were analyzed. Sixteen Vbeta gene segments were obtained accompanied by either Dbeta1 or Dbeta2 and by one of the nine Jbeta gene segments. Five different Cbeta cDNA sequences were obtained including four types of Cbeta1 sequences and one type of Cbeta2 sequence. The differences among the Cbeta1 sequences are either allelic polymorphisms or two splice variants, one being a product of exon1 splicing to exon3 (exon2 skipping), and another being an alternative splicing using a splice acceptor site newly discovered inside Cbeta1 exon4. The latter splice acceptor site was also found in human, mouse and horse all giving short cytoplasmic domain with Phe at their C-terminal ends. Other splicing products included trans splicing of Jbeta2 to Cbeta1, non-functional splicing of two Jbeta gene segments in tandem and a part of Jbeta2.7-Cbeta2 intron to Cbeta2 exon1. Numerous examples of splice variants may suggest the involvement of splicing in generating TCRbeta chain functional diversity. PMID- 17118452 TI - Strategy for affinity maturation of an antibody with high evolvability to (4 hydroxy-3-nitrophenyl) acetyl hapten. AB - In order to quantitate the contribution of amino acid replacements to an increase in affinity during affinity maturation, we measured thermodynamic parameters of the antigen-antibody interaction for a group of anti-(4-hydroxy-3-nitrophenyl) acetyl monoclonal antibodies whose differences in amino acid sequences had arisen only from somatic hypermutation. We prepared a common ancestor and hypothetical intermediate clones that might occur on the affinity maturation pathway, by employing site-directed mutagenesis. Isothermal calorimetric titration of the antigen-antibody reaction revealed that antibody evolution proceeds in two steps. The first step is driven by a decrease in enthalpy, in which two amino acid replacements in the VL region play an essential role. Further accumulation of amino acid replacements in VH and VL regions during the second step induce a progressive increase in affinity, which is driven by an increase in entropy, which has a cooperative mutational effect. PMID- 17118453 TI - IKK antagonizes CD95 ligation-mediated apoptosis by regulating NF-kappaB activity. AB - The CD95 (Apo1/Fas)/CD95 ligand system plays pivotal roles in various aspects of immune regulation and function by triggering apoptosis. Besides the apoptosis signaling pathway, CD95 ligation also induces the activation of NF-kappaB. Previous studies suggest that IkappaB kinase (IKK) may be a key player in cell survival by mediating NF-kappaB activation. However, the roles of IKK in CD95 ligation-mediated apoptosis and NF-kappaB activation are still not clear. In this report, we show that expression of the caspase-resistant uncleavable IKKbeta (UCIKKbeta) mutant suppressed CD95 ligation-mediated cell death in HeLa cells. Furthermore, CD95 ligation induced much more cell death in IKKbeta-/- murine embryonic fibroblasts (MEFs) than in wild type MEFs, despite that IKK was only marginally activated upon CD95 ligation. Pretreatment of HeLa cells with a specific IKK inhibitor NEMO-binding domain (NBD) peptide blocked CD95 ligation induced NF-kappaB transcriptional activity. And UCIKKbeta enhanced the basal NF kappaB activity, and consequently led to higher NF-kappaB activity upon CD95 ligation in HeLa cells. Therefore, IKK antagonizes CD95 ligation-mediated apoptosis by regulating NF-kappaB activity. PMID- 17118454 TI - Molecular cloning and characterization of Toll-like receptor 9 in Japanese flounder, Paralichthys olivaceus. AB - Toll-like receptor (TLR) 9 cDNA and gene were cloned from Japanese flounder, Paralichthys olivaceus. The Japanese flounder TLR9 cDNA encodes 1065 amino acids. The leucine-rich domain (LRD) and the Toll/interleukin-1 receptor (TIR) domain found in other vertebrate TLR9s were conserved in Japanese flounder TLR9. The gene is composed of three exons and two introns. The Japanese flounder tumor necrosis factor (TNF) gene promoter was activated in Japanese flounder TLR9 transformed hirame natural embryo (HINAE) cells upon stimulation with synthesized CpG oligodeoxynucleotide (ODN), but not by stimulation with GpC ODN. The Japanese flounder TLR9 gene was highly expressed in epithelial and lymphoid organs, such as the gills, intestines, kidney, spleen and stomach in an apparently healthy fish. The mRNA copy numbers of Japanese flounder TLR9 and its adapter protein, the myeloid differentiation factor 88 (MYD88) were increased in some organs including blood, gill, kidney and spleen after Edwardsiella tarda challenge. Immunohistochemical analysis revealed that TLR9 and MYD88 were expressed in the same cells of kidney. Few TLR9-expressing cells were found in gill, kidney and spleen in healthy Japanese flounder, but many were found in these organs after E. tarda challenge and were coincident with lesions that had been colonized by the bacteria. PMID- 17118455 TI - TRIM21 is a trimeric protein that binds IgG Fc via the B30.2 domain. AB - TRIMs comprise a large protein family that include anti-retroviral restriction factors such as TRIM5alpha. Auto-antibodies to TRIM21 (Ro52) are a common serological feature of patients with Sjogren's syndrome and systemic lupus erythematosus (SLE). We show that, in addition to this autoantibody response, TRIM21 binds specifically to the Fc region of human IgG isotypes 1, 2 and 4, via a conformation dependent interaction. The minimal binding epitope was identified as the C-terminal B30.2 domain. The interaction was independent of N-linked glycosylation of the IgG CH2 domain. TRIM21 formed a trimer that competed with protein A for binding to IgG Fc. We conclude that TRIM21 binds to the consensus CH2/CH3 domain interface in the Fc region, overlapping the binding site of several other proteins, including Staphylococcus aureus protein A and Streptococcus spp. protein G. The data suggest that the normal function of TRIM21 involves regulation of IgG functions and that TRIM/B30.2 molecules may have broader and unsuspected roles in innate immunity, beyond that of retroviral restriction. PMID- 17118457 TI - D-Aspartic acid: an endogenous amino acid with an important neuroendocrine role. AB - D-Aspartic acid (d-Asp), an endogenous amino acid present in vertebrates and invertebrates, plays an important role in the neuroendocrine system, as well as in the development of the nervous system. During the embryonic stage of birds and the early postnatal life of mammals, a transient high concentration of d-Asp takes place in the brain and in the retina. d-Asp also acts as a neurotransmitter/neuromodulator. Indeed, this amino acid has been detected in synaptosomes and in synaptic vesicles, where it is released after chemical (K(+) ion, ionomycin) or electric stimuli. Furthermore, d-Asp increases cAMP in neuronal cells and is transported from the synaptic clefts to presynaptic nerve cells through a specific transporter. In the endocrine system, instead, d-Asp is involved in the regulation of hormone synthesis and release. For example, in the rat hypothalamus, it enhances gonadotropin-releasing hormone (GnRH) release and induces oxytocin and vasopressin mRNA synthesis. In the pituitary gland, it stimulates the secretion of the following hormones: prolactin (PRL), luteinizing hormone (LH), and growth hormone (GH) In the testes, it is present in Leydig cells and is involved in testosterone and progesterone release. Thus, a hypothalamus-pituitary-gonads pathway, in which d-Asp is involved, has been formulated. In conclusion, the present work is a summary of previous and current research done on the role of d-Asp in the nervous and endocrine systems of invertebrates and vertebrates, including mammals. PMID- 17118458 TI - High-throughput protein digestion by trypsin-immobilized monolithic silica with pipette-tip formula. AB - Based on the monolithic silica gel materials with hierarchical pore structure and on the SPE devices (MonoTip) developed thereof, a trypsin-immobilized monolithic silica in a pipette tip (MonoTip Trypsin) suitable for digesting proteins has been newly developed. The surface of monolithic silica fixed into the tip was chemically modified with trypsin via an aminopropyl group. Trypsin-immobilized monolith successfully performed a rapid digestion of reduced and alkylated proteins with only a few times pipetting operation for the pre-treatment procedure of chromatographic analysis. The novel solid-phase digestion tool using monolithic silica allows a high-throughput trypsin proteolysis of bio-substances in proteomics. PMID- 17118456 TI - Transient receptor potential channels as novel effectors of brain-derived neurotrophic factor signaling: potential implications for Rett syndrome. AB - In addition to their prominent role as survival signals for neurons in the developing nervous system, neurotrophins have established their significance in the adult brain as well, where their modulation of synaptic transmission and plasticity may participate in associative learning and memory. These crucial activities are primarily the result of neurotrophin regulation of intracellular Ca(2+) homeostasis and, ultimately, changes in gene expression. Outlined in the following review is a synopsis of neurotrophin signaling with a particular focus upon brain-derived neurotrophic factor (BDNF) and its role in hippocampal synaptic plasticity and neuronal Ca(2+) homeostasis. Neurotrophin signaling through tropomyosin-related kinase (Trk) and pan-neurotrophin receptor 75 kD (p75(NTR)) receptors are also discussed, reviewing recent results that indicate signaling through these two receptor modalities leads to opposing cellular outcomes. We also provide an intriguing look into the transient receptor potential channel (TRPC) family of ion channels as distinctive targets of BDNF signaling; these channels are critical for capacitative Ca(2+) entry, which, in due course, mediates changes in neuronal structure including dendritic spine density. Finally, we expand these topics into an exploration of mental retardation (MR), in particular Rett Syndrome (RTT), where dendritic spine abnormalities may underlie cognitive impairments. We propose that understanding the role of neurotrophins in synapse formation, plasticity, and maintenance will make fundamental contributions to the development of therapeutic strategies to improve cognitive function in developmental disorders associated with MR. PMID- 17118459 TI - Monitoring synaptic transmission in primary neuronal cultures using local extracellular stimulation. AB - Various techniques have been applied for the functional analysis of synaptic transmission in cultured neurons. Here, we describe a method of studying synaptic transmission in neurons cultured at high-density from different brain regions such as the cortex, striatum and spinal cord. We use postsynaptic whole-cell recordings to monitor synaptic currents triggered by presynaptic action potentials that are induced by brief stimulations with a nearby extracellular bipolar electrode. Pharmacologically isolated excitatory or inhibitory postsynaptic currents can be reliably induced, with amplitudes, synaptic charge transfers, and short-term plasticity properties that are reproducible from culture to culture. We show that the size and kinetics of pharmacologically isolated inhibitory postsynaptic currents triggered by single action potentials or stimulus trains depend on the Ca2+ concentration, temperature and stimulation frequency. This method can be applied to study synaptic transmission in wildtype neurons infected with lentiviruses encoding various components of presynaptic release machinery, or in neurons from genetically modified mice, for example neurons carrying floxed genes in which gene expression can be acutely ablated by expression of Cre recombinase. The preparation described in this paper should be useful for analysis of synaptic transmission in inter-neuronal synapses formed by different types of neurons. PMID- 17118460 TI - Cloning, sequencing and expression of white rhinoceros (Ceratotherium simum) interferon-gamma (IFN-gamma) and the production of rhinoceros IFN-gamma specific antibodies. AB - Bovine tuberculosis (BTB) is endemic in African buffalo (Syncerus caffer) in the Kruger National Park (KNP). In addition to buffalo, Mycobacterium bovis has been found in at least 14 other mammalian species in South Africa, including kudu (Tragelaphus strepsiceros), Chacma baboon (Papio ursinus) and lion (Panthera leo). This has raised concern about the spillover into other potentially susceptible species like rhinoceros, thus jeopardising breeding and relocation projects aiming at the conservation of biodiversity. Hence, procedures to screen for and diagnose BTB in black rhinoceros (Diceros bicornis) and white rhinoceros (Ceratotherium simum) need to be in place. The Interferon-gamma (IFN-gamma) assay is used as a routine diagnostic tool to determine infection of cattle and recently African buffalo, with M. bovis and other mycobacteria. The aim of the present work was to develop reagents to set up a rhinoceros IFN-gamma (RhIFN gamma) assay. The white rhinoceros IFN-gamma gene was cloned, sequenced and expressed as a mature protein. Amino acid (aa) sequence analysis revealed that RhIFN-gamma shares a homology of 90% with equine IFN-gamma. Monoclonal antibodies, as well as polyclonal chicken antibodies (Yolk Immunoglobulin-IgY) with specificity for recombinant RhIFN-gamma were produced. Using the monoclonals as capture antibodies and the polyclonal IgY for detection, it was shown that recombinant as well as native white rhinoceros IFN-gamma was recognised. This preliminary IFN-gamma enzyme-linked immunosorbent assay (ELISA), has the potential to be developed into a diagnostic assay for M. bovis infection in rhinoceros. PMID- 17118461 TI - The involvement of mast cells and mast cell proteinases in the intestinal response to equine cyathostomin infection. AB - Cyathostomins (Cyathostominae) are regarded as the most pathogenic equine nematode worldwide. These nematodes are difficult to control in equine populations due to emerging anthelmintic resistance and evasion of encysted larval cyathostomins to regular modern anthelmintics. Mast cells and their proteinases have been shown to play a role in the mammalian immune response to nematode infections. Involvement of mast cells and mast cell proteinases in the equine immune response to cyathostomin infection is proposed. A technique was established to perform immunohistochemical staining using polyclonal rabbit anti equine mast cell proteinase-1 (eqMCP-1) and anti-equine tryptase on formalin fixed large intestinal sections, from horses classified as cyathostomin positive and negative at the time of death based upon larval enumeration. Quantitative analysis of antibody labelled mast cells was used to detect mast cell proteinases in equine large intestinal sections positive and negative for cyathostomin larvae. This demonstrated an increase in equine tryptase labelled mucosal and submucosal mast cells in cyathostomin positive horses. This study has established an immunohistochemical technique to demonstrate mast cell proteinases in formalin fixed large intestinal sections. This technique may be used to determine possible involvement of mast cells and their proteinases in the equine immune response to cyathostomin larvae. Further studies are required to define a specific role. PMID- 17118462 TI - Immunoblot analysis for IgE-reactive components of fetal calf serum in dogs that developed allergic reactions after non-rabies vaccination. AB - We previously demonstrated the presence of IgE directed to fetal calf serum (FCS) in the sera from dogs that developed allergic reactions after vaccination. In this study, by an immunoblot analysis, we investigated the IgE-reactive components of FCS using sera from 16 dogs that exhibited allergic reactions after vaccination. The immunoblot analysis revealed that several FCS proteins of approximately 25-, 50-, 66-, 75-, 120-, and 175-kDa strongly reacted with IgE in the sera from dogs that showed post-vaccination allergic reactions. The 66-kDa band was detected in the sera from 14 of the 16 dog serum samples analyzed in the immunoblot analysis for FCS, and it was speculated to be albumin based on its molecular weight; however, serum IgE reactivity to bovine serum albumin could be detected in only four of the 14 dog samples. These findings demonstrated that a variety of FCS components including albumin could function as allergens in dogs that developed allergic reactions after vaccination. PMID- 17118463 TI - Rapid identification of nontypeable and serotype b Haemophilus influenzae from nasopharyngeal secretions by the multiplex PCR. AB - OBJECTIVE: Heamophilus influenzae (H. influenzae) is an important pathogen responsible for both invasive and non-invasive infectious diseases. While encapsulated type b strain recognized as a major cause of severe invasive diseases, nontypeable strains are the major causes of non-invasive infectious diseases. Detection of this pathogen from nasopharyngeal secretions (NPS) is important. METHODS: We developed a multiplex polymerase chain reaction (PCR) for rapid identification of nontypeable and serotype b H. influenzae from nasopharyngeal secretions. RESULTS: A total 25 nasopharyngeal secretions were evaluated in this study. The multiplex PCR provided rapid and unequivocal results for determining either nontypeable or encapsulated typeable especially type b strains including a determination of beta-lactamase productions. CONCLUSION: The multiplex PCR based serotyping provided more reliable results than slide agglutination test (SAT) and is a valuable and expeditious method for identification of H. influenzae with determining capsular serotypes. PMID- 17118464 TI - TaqMan real-time PCR for detection of hepatopancreatic parvovirus from Australia. AB - Hepatopancreatic parvovirus is an emerging disease in crustacean aquaculture. Consequently, methods of detection are needed that enable the sensitive detection and confirmation of the virus better than currently used methods such as histology and conventional polymerase chain reaction (PCR). A TaqMan based real time PCR assay was developed for the detection of the Australian isolate of hepatopancreatic parvovirus which is only 85% similar to its nearest known relative. The TaqMan assay was developed within the capsid protein region of the genome and is optimised to detect as little as 10 copies of the targeted sequence per PCR vial. The hepatopancreatic parvovirus primers and probe were HPV140F 5' CTA CTC CAA TGG AAA CTT CTG AGC-3', HPV140R 5'-GTG GCG TTG GAA GGC ACT TC-3' and HPV140probe 5'-FAM TAC CGC CGC ACC GCA GCA GC TAMRA-3', respectively. The assay was specific for the hepatopancreatic parvovirus strain from Australian Penaeus merguiensis as it did not detect related crustacean and canine parvoviruses from Australia. In addition, the very low homology of the target sequence with published sequences from the Thai and Korean strains of hepatopancreatic parvovirus and other prawn viruses such as WSSV, suggested this assay would be specific for the Australian hepatopancreatic parvovirus isolate. Furthermore, it detected hepatopancreatic parvovirus in 22/22 wild-caught P. merguiensis clinical samples and 473/545 (87%) farmed P. merguiensis. This assay has the potential to be used for diagnostic purposes and in robotic applications, particularly for the detection and quantitation of low-grade infections. PMID- 17118465 TI - Design of a single tube RT-PCR assay for the diagnosis of human infection with highly pathogenic influenza A(H5) viruses. AB - Concerns about emergence of a pandemic strain of influenza have been increasing. The strains of highly pathogenic influenza A(H5N1) currently circulating are considered among the most plausible candidates for giving rise to a pandemic strain. In this study the design and development of a RT-PCR assay specific for these highly pathogenic influenza A(H5) strains is presented. This is achieved in part by the design of a primer targeting the coding region for the protease cleavage site of the hemagglutinin, and another primer derived from a pan hemagglutinin RT-PCR assay also presented in this study. It is shown that the HPAI A(H5) specific assay amplifies only the nucleic acids of highly pathogenic A(H5), with a high sensitivity. PMID- 17118466 TI - Quality assurance of genotyping array for detection and typing of human papillomavirus. AB - The aim of this study was to evaluate the sensitivity, specificity, reliability and reproducibility of the EasyChip HPV blot for human papillomavirus (HPV) genotyping. Type-specific sensitivity and specificity for 39 types of HPV (HPV 6, 11, 16, 18, 26, 31, 32, 33, 35, 37, 39, 42, 43, 44, 45, 51, 52, 53, 54, 55, 56, 58, 59, 61, 62, 66, 67, 68, 69, 70, 72, 74, 82, CP8061, CP8304, L1AE5, MM4, MM7 and MM8) were examined. The operating environment, reliability, reproducibility and blot interpretation were assessed by a quality assurance system. Each batch experiment contained samples from 89 cervical specimens and 7 extrinsic controls. Caski, HeLa and Jurkat cells, male human blood cell DNA and sterile water were used to assess reliability. Furthermore, pairs of sibling controls were used to assess reproducibility. The overall sensitivity of HPV detection was 1-50 copies of HPV genome equivalent. There was no cross-reactivity with amplicons of other HPV genotypes. One hundred batch experiments demonstrated that the reliability was excellent. The intra-batch and inter-batch reproducibility was 98 and 97%, respectively. It was concluded that the EasyChip HPV blot is a highly sensitive, reliable and reproducible tool for detection and identification of HPV genotypes. PMID- 17118467 TI - PAG mu opioid receptor activation underlies sex differences in morphine antinociception. AB - Given the findings that (1) systemic opioid antinociception varies by estrous stage in females and (2) the magnitude of sex differences in opioid antinociception is negatively correlated with opioid agonist efficacy, we hypothesized that sex differences in the function of the descending pain modulatory system are likely influenced by estrous stage in females and by the number of available opioid receptors therein. The present study tested these hypotheses by (1) comparing antinociception produced by morphine microinjection to the ventral periaqueductal gray (vPAG) in females at different stages of the estrous cycle and (2) examining systemic morphine antinociception in males versus females under conditions of reduced vPAG mu opioid receptor availability. When estrous stage of females was not controlled for (Experiment 1), there was no significant sex difference in tail withdrawal antinociception following morphine microinjection (0.3-10microg), although morphine was more potent in males than females in producing immobility. Experiment 2 showed that intra-vPAG morphine produced less antinociception and immobility in estrus than in diestrus females; that is, only estrus females' response to morphine was lower than that of males. Experiment 3 showed that microinjection of the irreversible mu opioid antagonist beta-funaltrexamine (beta-FNA) into the vPAG shifted the systemic morphine dose effect curve farther to the right in females than in males. That is, a reduction in available vPAG mu opioid receptors had a greater impact on opioid antinociception in females than in males, suggesting that females have fewer vPAG mu opioid receptors than males. Overall, these data suggest that ovarian hormones and PAG mu opioid receptor density contribute to sex differences in antinociception produced by morphine. PMID- 17118468 TI - Bioaccumulation and biotransformation of chiral triazole fungicides in rainbow trout (Oncorhynchus mykiss). AB - There are very little data on the bioaccumulation and biotransformation of current-use pesticides (CUPs) despite the fact that such data are critical in assessing their fate and potential toxic effects in aquatic organisms. To help address this issue, juvenile rainbow trout (Oncorhynchus mykiss) were exposed to dietary concentrations of a mixture of chiral triazole fungicides (bromuconazole, cyproconazole, metconazole, myclobutanil, penconazole, propiconazole, tebuconazole, tetraconazole, and triadimefon) and a chiral legacy pesticide [alpha-hexachlorocyclohexane (alpha-HCH)] to study the bioaccumulation and biotransformation of these CUPs. Fish accumulated all triazoles rapidly during the 8 day uptake phase, and was followed by rapid elimination, which was estimated by taking accelerated sampling times during the 16 day depuration period. Half-lives (t1/2s) and times to 95% elimination (t95s) ranged from 1.0 to 2.5 and 4.5 to 11.0 days, respectively. Chiral analysis suggested no significant selectivity in biotransformation for most of the compounds based on statistically unaltered enantiomer fractions (EFs) in the fish compared to food values; exceptions were a change in EF of myclobutanil and changes in diastereomer fractions (DFs) of propiconazole and cyproconazole. No biotransformation was observed for alpha-HCH based on consistent EFs in the fish throughout the experiment and a t1/2 (15.8 days) that fell within the 95% confidence interval of a log K(ow)-log t1/2 relationship developed for assessing biotransformation of organic contaminants. This relationship did show that biotransformation accounted for the majority (ranging from 59.9 to 90.4%) of the elimination for all triazoles, and that triazole compounds with oxygen and hydroxyl functional groups were more easily biotransformed. This research indicated that chiral analysis may potentially miss biotransformation of CUPs and other potential non-persistent organic contaminants and shows the utility of the log K(ow)-log t1/2 relationship as a mechanistic tool for quantifying biotransformation. Based on the rapid biotransformation of the triazoles, future research should focus on formation of metabolites and their fate and possible effects in the environment. PMID- 17118469 TI - Receptor-binding studies of the DBLgamma domain of Plasmodium falciparum erythrocyte membrane protein 1 from a placental isolate. AB - We have previously identified a number of DBLgamma domains in Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) transcripts obtained from placental parasite isolates, showing that they bind specifically to chondroitin sulfate A (CSA) (Khattab A, Kun J, Deloron P, Kremsner PG, Klinkert MQ. Variants of Plasmodium falciparum erythrocyte membrane protein 1 expressed by different placental parasites are closely related and adhere to chondroitin sulfate A. J Infect Dis 2001;183:1165-9). Here we give a more detailed physico-chemical and binding characterisation of the soluble, recombinant DBLgamma domain derived from one of these isolates. Results from circular dichroism and limited proteolysis experiments are consistent with the recombinant domain being expressed with the native fold. Specific binding of DBLgamma to placental cryosections was demonstrated by labeling with antibodies raised against the recombinant domain; binding was diminished after treatment of the cryosections with chondroitinase or by blocking with anti-CSA antibody, showing that CSA mediates the interaction. Binding of the DBLgamma domain to purified placental chondroitin sulfate proteoglycan (CSPG) was also studied using surface plasmon resonance techniques, with DBLgamma as analyte and CSPG immobilised on the sensor chip; these quantitative measurements gave an affinity constant in the mu-molar range under the conditions used. The native conformation of the DBLgamma domain is essential for CSPG recognition since binding to the sensor chip is abolished when the protein is irreversibly reduced. As with the placental cryosections, association was significantly reduced after treating the immobilised CSPG with chondroitinase. Together, these results demonstrate specific interaction between the DBLgamma domain and the placental receptor. PMID- 17118470 TI - The subcellular localisation of trypanosome RRP6 and its association with the exosome. AB - The exosome, a complex of 3'-exoribonucleases and associated proteins, is involved in the degradation of eukaryotic mRNAs in the cytoplasm, and has RNA processing and quality control functions in the nucleus. In yeast, the nuclear exosome differs from the cytoplasmic one in that it contains an additional non essential component, Rrp6p. In contrast, a small proportion of human RRP6 has been shown to localise to the cytoplasm as well. When we purified the Trypanosoma brucei exosome from cytosolic extracts we found RRP6, apparently in stoichiometric amounts. We here confirm that RRP6 is in the trypanosome cytoplasm and nucleus. The level of RRP6 was unaffected by depletion of core exosome subunits by RNA interference and over-expression of tagged RRP6 was possible, indicating that RRP6 can be present independent of exosome association. PMID- 17118471 TI - mRNA made during heat shock enters the first round of translation. AB - To determine whether mRNA synthesized during a heat shock is translated at least once in spite of the strong inhibition of translation by heat shock, we used nonsense-mediated decay (NMD) as an assay since NMD requires a round of translation. As NMD substrate we used the human psigammaE-crystallin gene, which contains a premature termination codon, and as control, its close relative, the human gammaD-crystallin gene, both placed under control of the Hsp70 promoter. We show that no spliced psigammaE-crystallin mRNA can be detected in heat shocked cells, suggesting that NMD resumes as soon as splicing is restored. We further show that newly synthesized mRNAs co-sediment with the 40S ribosomal subunits, indicating that the transcripts are recruited to the translation machinery but are stalled at the translation initiation stage. Using fluorescence loss in photobleaching (FLIP) we show that cytoplasmic EGFP-CBP20 is immobile in heat shocked cells. CBP20 is part of the cap binding complex which is thought to direct the first round of translation. Together our data suggest that all mRNAs made during heat shock enter the pioneer round of translation. PMID- 17118472 TI - Relationship between balance performance and musculoskeletal pain in lower body comparison healthy middle aged and older adults. AB - The purpose of this study was to determine relationship between balance performance and pain in lower body among healthy adults. Two hundred and forty volunteer subjects (125 middle aged and 115 elderly people) aged 50 years and above participated. The average age was 61.52+/-8.22 years (range 50-75 years). The functional reach test (FRT) was used to measure balance ability. The visual analog scale (VAS) was used to measure pain intensity. Subjects were also asked to indicate sites they experienced pain in their lower body (e.g., low back, hip, knee, ankle, toes). Falls history was also recorded. The elderly people had a lower FRT score as compared to the middle-aged adults (p<0.001). Knee, low back, and hip pain were most common in both groups. The elderly people reported more frequency of falls than the middle-aged adults. The score of the FRT was higher among men (20.67+/-7.16cm) than among women (18.77+/-6.59cm). In both groups; the women had a higher VAS score than those of matched the men. Although the middle aged adults had higher and better scores than the elderly people, the data showed that the middle-aged adults are also at risk and should be prepared properly for healthy aging. PMID- 17118473 TI - The association between Mycobacterium avium subsp. paratuberculosis fecal shedding or clinical Johne's disease and lactation performance on two Minnesota, USA dairy farms. AB - Lactation performance of cows infected with Mycobacterium avium subsp. paratuberculosis (Map) was previously studied using only serum ELISA as a diagnostic method. This study evaluated on two dairy farms in Minnesota, USA the lactation performance (measures of health, production, reproduction, and survival) of cows shedding Map in feces before calving and of cows culled with clinical signs consistent with Johne's disease (JD) during the subsequent lactation. Fecal samples were collected from 1052 cows within 21 day before calving and tested for Map with bacterial culture. Producers' observed signs of clinical disease (milk fever, retained placenta, metritis, ketosis, displaced abomasum, lameness, mastitis, pneumonia, and JD) and production and reproduction data were recorded for each cow. The association between fecal shedding or clinical JD and lactation performance was evaluated. Logistic regression was used to evaluate the association with any clinical and subclinical diseases as the outcome. General linear model was used to evaluate the association with milk production, and survival analysis techniques were used to evaluate the association with days in the study before culling and days from calving to conception. In 84 cows (8% of 1052 cows) fecal samples were positive for Map (46% light, 26% moderate, and 28% heavy shedders). In multivariable analysis, light, moderate, and heavy fecal shedding cows produced on average 537, 1403, and 1534 kg, respectively, less milk per lactation and 1.4, 5.2, and 7.5 kg, respectively, less milk per day than fecal negative cows. Fecal culture positive cows were less likely to be bred and conceive. In the multivariable analysis the 56 cows culled with presumed JD produced approximately 1500 kg/lactation or 5 kg/day less than all other cows. The negative economic impact implied by decreased lactation performance in cows shedding Map or with clinical JD may motivate producers to implement programs to control Map infection and subsequent JD. PMID- 17118474 TI - Cloning and expression of a DNA ligase from the hyperthermophilic archaeon Staphylothermus marinus and properties of the enzyme. AB - The gene encoding Staphylothermus marinus DNA ligase (Sma DNA ligase) was cloned and sequenced. The gene contains an open reading frame consisting of 1836bp, which encodes for 611 amino acid residues. Upon alignment of the entire amino acid sequence, Sma DNA ligase showed a high degree of sequence homology with the hyperthemophilic archaeal DNA ligases, 67% identity with Aeropyrum pernix K1, and 40% identity with both Pyrococcus abyssi and Thermococcus kodakarensis. An extremely high sequence identity was observed in the six conserved motifs indicative of DNA ligase. The Sma DNA ligase gene was expressed under the control of the T7lac promoter on the pET-22b(+) in Escherichia coli BL21-CodonPlus(DE3) RIL. The expressed enzyme was then purified by heat treatment followed by ion exchange and metal affinity column chromatography. The enzyme was activated by both Mg(2+) and Mn(2+), and its activity was inhibited by Ca(2+) and Zn(2+). Sma DNA ligase can utilize both ATP and ADP as cofactors. The half-life of the enzyme at 100 degrees C was determined to be approximately 2.8h. The enzyme catalyzed cohesive-end intramolecular and intermolecular joining and blunt-end intermolecular joining in the presence of tricine-NaOH buffer and Mn(2+), using either ATP or ADP. PMID- 17118475 TI - Limiting factors governing protein expression following polyethylenimine-mediated gene transfer in HEK293-EBNA1 cells. AB - Transient gene expression in mammalian cells is intensively used for the rapid generation of recombinant proteins for biochemical, biophysical and pre-clinical studies. Still, the principles behind DNA transfer to the cells and the cellular cascade of events that ultimately dictate protein expression levels are not fully understood. Using polyethylenimine (PEI) mediated transfection of HEK293-EBNA1 cells, we sought to determine the most critical parameters that drive and limit recombinant protein production. Our results showed that a maximum of 65,000 plasmid copies/cell can be recovered in total extracts at 1 day post transfection. Analyses performed after cell sorting revealed equal amounts of plasmid DNA in GFP-positive and -negative populations. However, nuclear plasmid content was three-fold higher in GFP-positive cells (1850 copies) than in GFP negative cells (550 copies). The fact that significant amounts of plasmid DNA are found in the nucleus of GFP-negative cells suggests that its transcriptional competency is impaired. Interestingly, transfecting cells using a wide range of plasmid quantities at the optimal DNA:PEI ratio did not significantly affect the number of expressing cells. Thus, it appears that successful transgene expression is more likely to depend on a cellular "competent" state than to the quantity of plasmid DNA delivered per cell. Moreover, Northern blot analysis and SEAP/GFP measurement following plasmid titration experiments showed that transcriptional and translational processes are operating near to saturation under optimal transfection conditions. Overall, our results suggest that events that regulate nuclear translocation of plasmid DNA and its transcriptional competency as well as translational/post-translational limitations represent major bottlenecks in the success of a PEI-mediated protein production. PMID- 17118476 TI - PEGylated lysine dendrimers for tumor-selective targeting after intravenous injection in tumor-bearing mice. AB - In this study, we synthesized a sixth generation lysine dendrimer (KG6) and two PEGylated derivatives thereof and evaluated their biodistribution characteristics in both normal and tumor-bearing mice. The intact KG6 showed a rapid clearance from the blood stream and non-specific accumulation in the liver and kidney. In contrast, the PEGylated derivatives showed a better retention in blood and low accumulativeness in organs dependent of the rate of PEGylation. In addition, PEGylated KG6 with a high modification rate was accumulated effectively in tumor tissue via the enhanced permeability and retention (EPR) effect. Moreover, we clarified that multiple administrations did not affect the biodistribution characteristics of a second dose of PEGylated KG6. PEGylated lysine dendrimer would be a useful material for a clinically applicable tumor-targeting carrier. PMID- 17118477 TI - Prevalence of the metabolic syndrome among Korean adults using the new International Diabetes Federation definition and the new abdominal obesity criteria for the Korean people. AB - This study was performed to compare the prevalence of the metabolic syndrome according to the International Diabetes Federation (IDF) and National Cholesterol Education Program (NCEP) definitions, and abdominal obesity criteria of WHO and the Korean Society for the Study of Obesity (KSSO) in Korean adults. A total of 4452 adults aged > or =20 years from the Korean National Health and Nutrition Examination Survey 2001 were analyzed. The prevalence of the metabolic syndrome estimated by NCEP definition with WHO criteria, NCEP with KSSO, IDF with WHO, and IDF with KSSO were 26.7%, 23.7%, 23.8% and 17.5%, respectively. The agreement percent among the four definitions ranged from 88.7% to 100% in men, and from 85.6% to 94.9% in women. The NCEP-defined metabolic syndrome was more strongly associated with hypertension and diabetes than the IDF-defined metabolic syndrome (age-adjusted odds ratio: 5.1 versus 3.6 for hypertension and 6.4 versus 3.2 for diabetes in men, respectively; 5.4 versus 3.4-4.3 for hypertension and 11.1 versus 3.8-4.2 for diabetes in women, respectively). Both definitions of the metabolic syndrome were associated with coronary heart disease or stroke only in women. Prospective studies are warranted to evaluate the predictive ability of the new definition of the metabolic syndrome and the new criteria of abdominal obesity for cardiovascular morbidity and mortality in Korean adults. PMID- 17118478 TI - Undertreatment of cardiovascular risk factors among persons with diabetes in the United States. AB - OBJECTIVE: We examined the extent of control of cardiovascular risk factors and distance from goal for those with uncontrolled levels in a recent sample of U.S. adults with diabetes. METHODS: In the cross-sectional National Health and Nutrition Examination Survey 2001-2002, 532 (projected to 15.2 million) or 7.3% of adults aged >/=18 years had diabetes. Use of antihypertensive, antidiabetic and antidyslipidemic medications was examined. We determined the proportion of subjects not at goal for blood pressure (BP), lipids and glycosylated hemoglobin (A1C) and examined the distance from goal for those not under control. RESULTS: Overall, 50.2% of subjects with diabetes were not at goal for A1C, 64.6% for low density lipoprotein-cholesterol (LDL-C), 52.3% for high density lipoprotein cholesterol (HDL-C), 48.6% for triglycerides and 53.0% BP. Only 5.3% of men and 12.7% of women with diabetes were simultaneously at goal for A1C, LDL-C and BP. Even among those on treatment, most were not at goal for these parameters. Women were more likely to have LDL-C and HDL-C not at goal than men. Non-Hispanic Blacks were more often not at goal for BP and LDL-C. Mean distances from targets were 36mg/dL for LDL-C, 18mmHg for systolic BP, 6mmHg for diastolic BP and 2.0% for A1C in patients not at goal. CONCLUSIONS: Many U.S. adults with diabetes have sub-optimal control of cardiovascular risk factors and remain far from target goals for BP, lipids and A1C, even if on treatment. PMID- 17118479 TI - Childhood obesity and its relation to serum adiponectin and leptin: a report from a population-based study. AB - This study examined the relationships between serum adiponectin (AD) and leptin (LP) levels, and obesity using a population-based cohort consisted of 315 (9-10 year olds: G1) and 308 (12-13 year olds: G2) school children. Serum AD, LP and other markers were compared according to the presence of obesity. The prevalence rates of obesity were 14.9% in G1 and 9.4% in G2. The medians of serum AD (microg/dl: non-obese/obese) were statistically lower in obese children (9.6/8.3 in G1, p<0.05; 8.9/6.6 in G2, p<0.05), and the medians of serum LP (ng/dl) were statistically higher in obese children (3.7/12.5 in G1, p<0.05; 2.9/8.4 in G2, p<0.05). The serum LP levels were significantly positively correlated with percent overweight (POW) irrespective of age and sex, and the serum AD levels were significantly negatively correlated with POW except for boys in G1. Multivariate regression analyses revealed that LP, LDL-cholesterol and gender in G1, and LP, AD, blood pressure and gender in G2 were significantly correlated with POW. A large-scale, population-based study revealed that AD was lower and LP higher in obese children, and that the obese status in G2 was related to a worse metabolic profile than the case in G1. PMID- 17118480 TI - Association of sulfonylurea receptor 1 genotype with therapeutic response to gliclazide in type 2 diabetes. AB - To investigate the effects of sulfonylurea receptor 1 (SUR1) exon 33 (TCC-->GCC, S1369A) polymorphism on responsiveness to gliclazide. About 115 patients with type 2 diabetes were treated with gliclazide for 8 weeks. SUR1 genotypes were tested by Taqman-PCR. After gliclazide treatment, there was association between T/G polymorphism and decrease of HbA1c. G carriers were more sensitive to gliclazide and the decrease of HbA1c was more significant than TT genotype (TT, 0.76%+/-1.70%; TG+GG, 1.60%+/-1.39%, P=0.044). The polymorphism of SUR1S1369A was associated with the therapeutic efficacy of gliclazide. PMID- 17118481 TI - Viral/self-mimicry and immunological cross-reactivity as a trigger of hepatic C virus associated autoimmune diabetes. PMID- 17118482 TI - Drug use and health-seeking behavior for childhood illness in Vietnam--a qualitative study. AB - OBJECTIVES: To describe and understand health-seeking behavior and drug use among caregivers, in particular antibiotics, for childhood illnesses in Vietnam. METHODS: A total of six focus group discussions were conducted with child care providers. A thematised interview guide was used and qualitative content analysis was applied. Meaning units in the texts dealing with the same content material were identified, condensed and compared until several emerging themes were found. Finally, similarities and differences in the views between rural and urban groups were described. RESULTS: Self-treatment and self-medication were prominent in the urban groups, whereas compliance and trust in doctors were more common among the rural participants. Misuse and misconceptions regarding the properties of drugs were reported leading to worries about irrational drug use in the community. Despite the widespread use of private services in both areas, the public health service was reported to be the most trusted recourse for serious cases. CONCLUSIONS: Qualitatively different ways of using drug and health care services are reported for childhood sickness in rural and urban Vietnam. These results provide rich information for the development of quantitative studies on drug use for children and point to the need to involve all health facilities in consideration. PMID- 17118483 TI - Morphometric analysis of face in dysmorphology. AB - Dysmorphology refers to study of human congenital malformations (birth defects). Most of the case reporting in dysmorphology is subjective and is based on experience of the reporting clinician. We have used the methods of geometric morphometrics to analyze the variation in faces of normal individuals and those with dysmorphic syndrome. We obtained photographs of 20 individuals with Rubinstein Taybi syndrome and 30 normal, age and sex matched individuals. The photographs were digitized with 16 landmarks on the face to obtain 32 "x" and "y" co-ordinates. These co-ordinates were then subjected to generalized procrustes superimposition in order to normalize for effects of size, rotation and position of image. The procrustes residuals thus obtained were then subjected to principal component analysis. The principal component analysis resulted in extraction of three important principal components explaining 41%, 17% and 14% of variance, respectively. Discriminant analysis could differentiate the two groups using first two principal component scores for each individual, with a predictive accuracy of 76% (Wilks lambda=0.725, chi2=15.09, d.f.=2, p=0.001). Binary logistic regression analysis showed predictive accuracy of 78% based on this model. The utility of the subjective evaluation of facial characteristics is multifold. The results of the analysis can be used as representatives of the facial dysmorphism for any genotype-phenotype association study. We conclude that application of the principles of geometric morphometrics to study of shape variation in facies of patients with dysmorphic syndromes appears to be a promising new area of research. PMID- 17118484 TI - Virus-based nanoparticles (VNPs): platform technologies for diagnostic imaging. AB - Non-invasive imaging holds great promise for the early detection and treatment of human disease. The ability to both detect and follow disease processes or anatomical defects without biopsy, surgery, or other invasive techniques should lead to lower costs and higher quality of life. The use of "smart" nanoparticles, that combine multiple functions of targeting, imaging, and drug delivery, have tremendous potential to increase the sensitivity and specificity of therapies. These will facilitate early detection and reduce adverse side effects of treatment. There are many different classes of nanoparticles in development including dendrimers, liposomes, paramagnetic nanoparticles, and quantum dots, to name just a few. Here we focus on virus-based nanoparticles (VNPs) as platforms for the development of tissue-specific targeting and imaging agents in vivo. PMID- 17118485 TI - Freeze-drying of nanoparticles: formulation, process and storage considerations. AB - Freeze-drying has been considered as a good technique to improve the long-term stability of colloidal nanoparticles. The poor stability in an aqueous medium of these systems forms a real barrier against the clinical use of nanoparticles. This article reviews the state of the art of freeze-drying nanoparticles. It discusses the most important parameters that influence the success of freeze drying of these fragile systems, and provides an overview of nanoparticles freeze drying process and formulation strategies with a focus on the impact of formulation and process on particle stability. PMID- 17118486 TI - Toxin entry and trafficking in mammalian cells. AB - There is a vast number of bacterial and plant toxins that affect cytosolic targets in mammalian cells, and whether the purpose of the toxin is to act as a defence mechanism against predators, or to cause deliberate cell death in order to form an environment more suitable for bacterial growth, each of these toxins must cross a lipid membrane barrier in order to exert their effect. This review looks at the methods employed by bacterial and plant toxins in order to reach their target. We examine the trafficking methods involved in toxin transport throughout the cell, highlighting the proteins necessary for the toxins movement, and noting how many of the toxins hijack the cells own trafficking and protein processing machinery in order to reach their goals. Studying the trafficking of toxins has led to a greater understanding of retrograde transport, a process which has key relevance to the correct intracellular delivery of pharmacological agents. PMID- 17118487 TI - Multiple lung adenocarcinomas and AAH treated by surgical resection. AB - Atypical adenomatous hyperplasia (AAH) is often found in the lungs of patients with multiple primary lung adenocarcinoma; however, treatment for such patients has not been clearly defined. This report presents a case of multiple primary lung adenocarcinoma with multiple AAH treated by surgery. A 55-year-old man underwent two operations in 3 years and 8 months. Thirteen tumorous lesions were resected; 10 lesions were diagnosed as primary lung adenocarcinoma and the others as AAH. Two years and 6 months after the second operation, radiographic examination showed no evidence of tumor recurrence. The clinical course indicates that surgery can be a treatment strategy for synchronous or metachronous lung carcinoma with AAH. PMID- 17118488 TI - Increased expression and activity of repair genes TDP1 and XPF in non-small cell lung cancer. AB - Resistance to camptothecin (CPT), a topoisomerase I (Top1) inhibitor, is frequently encountered in non-small cell lung cancer (NSCLC) and CPT resistance is linked with TDP1, an enzyme capable of cleaving the covalent linkage between stabilized Top1 with DNA. The aim of this study is to evaluate the in vivo expression level of TDP1, as well as parallel repair pathway components XPF and MUS81, in primary NSCLC. We collected 30 un-matched and 4 NSCLC samples matched with normal lung tissue and 8 samples of non-neoplastic lung tissue from patients with and without lung cancer, and determined the protein expression of these three genes using Western blot and TDP1 activity by a specific enzymatic assay. Both TDP1 and XPF were overexpressed in over 50% of NSCLC tissues, with wide ranges of expression levels. MUS81 did not exhibit alteration in expression. Overexpression of TDP1 and XPF is common in NSCLC, and is therefore of interest as a possible contributor to drug resistance in NSCLC. PMID- 17118489 TI - Iodopovidone: an inexpensive and effective agent for chemical pleurodesis. PMID- 17118490 TI - Second-line treatment for advanced non-small cell lung cancer: how to design a clinical trial for a new agent? PMID- 17118491 TI - Short-term effects of chewing gum on snack intake and appetite. AB - Consumers report that chewing gum can reduce cravings and the likelihood of snacking. The present study set out to examine the effects of chewing gum on subjective appetite and snack energy intake (EI) in 60 participants (40 females, 20 males, 21.7+/-4 years; BMI=22.7+/-3.4) who came to the laboratory four times for lunch and then returned 3 h later for a snack. Participants consumed salty or sweet snacks after chewing gum (sugar-free or regular) for 15 min hourly after lunch or had no-gum. Hunger, desire to eat and fullness were rated immediately after lunch (T0) and hourly post-lunch (T1 and T2) until just before snack (T3). Chewing gum reduced EI by 36 cal (401.8+/-22 kcal) compared to no-gum (437.7+/-23 kcal; p=0.04). Rated hunger increased from T0 to T3 (p<0.001); however, this was less after gum compared to no-gum (p<0.01). Desire to consume salty and sweet snacks also increased. However, desire to eat sweet snacks (but not salty) increased less after gum compared to no-gum (p=0.004). Therefore, chewing gum suppressed appetite, specifically desire for sweets and reduced snack intake. This supports anecdotal reports by consumers and could inform weight control strategies. PMID- 17118493 TI - Pupillary cholinergic hypersensitivity predicts cognitive decline in community dwelling elders. AB - Previous research demonstrated that it is possible to distinguish patients with probable Alzheimer's disease from age-matched controls based on an exaggerated pupil dilation response to dilute tropicamide. The research reported here employed a prospective longitudinal design to follow over time (2-4 years) a sample of 55 community dwelling elders with and without an exaggerated pupil response using the pupil assay and a comprehensive battery of neuropsychological tests sensitive to pre-clinical AD. Discrete time survival modeling was used to assess the ability of the assay to predict a pattern of cognitive decline consistent with early AD. Analysis showed that there is an increased risk (odds ratio of 3) with a hypersensitive pupil response (>/=13% increase in pupil diameter over baseline diameter) for developing significant cognitive impairment in areas of memory attention and language in a pattern, consistent with pre clinical Alzheimer's disease. When controlling for ApoE allele type the odds ratio for pupil response as a risk factor increased to 4. The analysis also found that an exaggerated pupil response was a significant (p=.02) predictor of cognitive decline. This analysis of longitudinal data has shown that over time an exaggerated response on the pupil assay is a significant independent risk factor for developing pre-clinical Alzheimer's disease. The risk for developing pre clinical Alzheimer's disease is increased four-fold. PMID- 17118492 TI - Effects of melatonin and age on gene expression in mouse CNS using microarray analysis. AB - The expression levels of a number of genes associated with inflammation and immune function change with advancing age. Melatonin modulates gene expression levels of several of these genes. Therefore the declining levels of melatonin associated with age may play a role in the physiological effects of aging. We used oligonucleotide microarrays to measure age-related changes in mRNA expression in the murine CNS, and to study the effect of prolonged administration of dietary melatonin upon these changes. CB6F1 male mice were fed 40 ppm melatonin for 2.1 months prior to sacrifice at age 26.5 months, and compared with both age-matched controls and young, 4.5-month-old untreated controls. Total RNA was extracted from whole brain (excluding cerebellum and brain stem) and individual samples were hybridized to Affymetrix Mouse 430-2.0 arrays. The expression of a substantial number of genes was modulated by melatonin treatment and changes in selected genes were validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR). A subset of these genes did not change with age. Conversely, some genes modulated by age were also modulated by melatonin treatment. In general, melatonin treatment drove the expression levels of these genes closer to the expression levels detected in the younger animals. Notably, the abundance of lipocalin 2 (Lcn2) mRNA increased with age and was decreased in old animals treated with melatonin. Lcn2 is a member of the acute phase response family of proteins and its mRNA levels in the brain increase in response to inflammation. Many of the genes with expression reduced by melatonin are involved in inflammation and the immune system. This suggests that melatonin treatment may influence the inflammatory responses of old animals, driving them to resemble more closely those occurring in young animals. PMID- 17118494 TI - 3D-QSAR study of sulfonamide inhibitors of human carbonic anhydrase II. AB - 3D-QSAR models of Comparative of Molecular Field Analysis (CoMFA) and Comparative of Molecular Similarity Indices Analysis (CoMSIA) of 61 potent carbonic anhydrase II (CAII) sulfonamide inhibitors were performed using two methods. The conventional ligand-based 3D-QSAR studies were performed based on the lower energy conformations employing database alignment rule. The receptor-based 3D QSAR models were also derived using bioactive conformations obtained by docking compounds to the active sites of CAII. The receptor-based model gave q(2) values of 0.623 and 0.562, r(2) values of 0.986 and 0.987 for CoMFA and CoMSIA, respectively, which were much better than those of ligand-based model (q(2) values of 0.532 and 0.466). The predictive ability of the models was validated using the test set of 10 compounds that were not included in the training set of 51 compounds. Results of CoMFA and CoMSIA suggested that heterocyclic sulfonamides are more active than aromatic sulfonamides, in the latter 1,3,5 triazole group substituting one hydrogen atom of the amido is favored and moderate groups in its 4- and 6-position are required. These results provided further understanding of the relationship between the structural features of CAII and its activities, which should be applicable to design and find new potential CAII inhibitors. PMID- 17118495 TI - [An infiltrated eyelid]. PMID- 17118496 TI - The effect of education on hypothetical and actual oxygen administration decisions. AB - AIM: This study examined the effect of an education intervention on emergency nurses' decisions related to oxygen administration. METHOD: A controlled pre test/post-test quasi-experimental design was used. The intervention was a written self directed learning package. Outcome measures were (i) factual knowledge measured using parallel form multiple choice questions (MCQs) and (ii) clinical decisions measured using parallel form MCQs, parallel form patient scenarios and clinical practice observation. RESULTS: Eighty-eight nurses from 4 Melbourne EDs participated in the study (control group: n=37 and experimental group: n=51). Subgroups of nurses from the experimental group also participated in the patient scenarios (n=20) and clinical practice observation (n=10). Emergency nurses' knowledge increased as a function of education. Both patient scenario data and clinical practice observation showed decreased selection of nasal cannulae, increased selection of air entrainment masks and a trend towards selection of higher oxygen flow rates following education. CONCLUSIONS: Evaluation of educational interventions in nursing should focus on identifying strategies that enhance learning in a clinical environment, are valid in terms of the clinical context and culture in which they are being used and most importantly, produce sustained improvements in actual clinical practice. PMID- 17118497 TI - Enhancement of BCG-induced Th1 immune response through Vgamma9Vdelta2 T cell activation with non-peptidic drugs. AB - Since drug-activated gammadelta T cells promote dendritic cell (DC) maturation, we analyzed the effect of combining gammadelta T cell specific drugs with BCG in vitro. BCG-induced DC maturation was increased by bromohydrin-pirophosphate (BrHPP) or zoledronate (Zol)-activated gammadelta T cells. Specifically, the co culture with activated Vgamma9Vdelta2 T cells with BCG-infected DC resulted in a significant increase of the expression of CD80, CD86, CD40 and CD25 molecules on DC. Moreover, DC were able to produce increased levels of TNF-alpha and synthesize ex novo IL-15 without altering the IL-10/IL-12 immunoregulatory pathway. Finally, the Th1 immunity induced by BCG-infected DC on naive CD4 T cells was increased by gammadelta T cell activation with BrHpp or Zol. These data indicate that gammadelta T cell triggering drugs could be used to enhance the BCG induced Th1 immunity. PMID- 17118498 TI - A feminist exploration of Traveller women's experiences of maternity care in the Republic of Ireland. AB - OBJECTIVE: to explore Traveller women's experiences of maternity care so that their needs might be identified and perceptions of helpful responses elicited. DESIGN: feminist research methodology using unstructured non-directive interviews. SETTING: community setting in the Republic of Ireland. PARTICIPANTS: snowball sample of 13 Traveller women aged 19-42 years. Each had experienced between two and eight pregnancies. FINDINGS: a fluid concept of culture affected Traveller women's expectations and negotiation of maternity care. The essential nature of familism, socialisation and religious beliefs, and the particular emphasis placed upon peer support, reflected cohesive and supportive aspects of culture. The concept of 'possessive individualism' was portrayed as conflicting with contraceptive use, the uptake of preventive care and women's reporting of mental-health problems. Majority norm expectations of breast feeding, husband participation and 'rooming in' were culturally unacceptable. Political and structural factors, such as the direct discriminatory barriers created by general practitioner services, indirect discrimination arising from dysfunctional communication and control of information, poor housing and lack of public transport were the basic causes of inequity of access to care. IMPLICATIONS FOR PRACTICE: midwives are challenged to respond sensitively to cultural preferences, meet informational needs and act as political advocates in efforts to improve maternity care experiences for Traveller women. PMID- 17118499 TI - Dynamic measurement of patellofemoral contact pressure following reconstruction of the medial patellofemoral ligament: an in vitro study. AB - BACKGROUND: Surgical reconstruction of the medial patellofemoral ligament used to stabilize the patella against lateral dislocation may concomitantly produce alteration of the patellofemoral contact pressure distribution. Two different tendon transfer techniques of reconstructing the medial patellofemoral ligament, one dynamic and one static, as well as a proximal soft tissue realignment of the patella were investigated. METHODS: Eight human knee specimens were mounted in a kinematic knee simulator and isokinetic extension motion was simulated. Patellofemoral pressure was measured using a pressure sensitive film while a 100 N laterally directed dislocation load was applied to the patella. The specimens were evaluated in a physiologic state, as well as after dynamic reconstruction of the medial patellofemoral ligament using a distal transfer of the semitendinosus tendon, following static reconstruction using a semitendinosus autograft, and following proximal soft tissue realignment of the patella. FINDINGS: Following both reconstruction techniques of the medial patellofemoral ligament patellofemoral contact pressure was not significantly (P=0.49) altered. In contrast, after proximal realignment a trend (P=0.07) towards higher contact pressure near knee extension was observed. In the absence of a lateral dislocation load dynamic and static reconstruction resulted in a medialization (P=0.04) of the center of pressure, whereas under the application of a 100 N dislocation load the center of pressure showed no significant alteration. Following proximal realignment the center of pressure was significantly medialized without (P<0.01) and with a dislocation load (P=0.01) throughout the entire range of knee motion. INTERPRETATION: Static and dynamic ligament reconstruction of the medial patellofemoral ligament did not alter patellofemoral pressure. Proximal realignment, on the other hand, resulted in a constant medialization of the patellofemoral pressure. The data suggest that the reconstruction techniques would be associated with a low risk of causing premature cartilage degeneration due to excessive patellofemoral contact pressure, whereas proximal realignment could cause medial overload of the patellofemoral joint. PMID- 17118500 TI - Nonlinear finite element analysis for musculoskeletal biomechanics of medial and lateral plantar longitudinal arch of Virtual Chinese Human after plantar ligamentous structure failures. AB - BACKGROUND: Musculoskeletal diseases of the foot such as stress fractures, tendonitis and subsequent pain are commonly associated with elevated stresses/strains of abnormal plantar arch after plantar ligamentous structure failures. The goal of this study was to develop anatomically detailed, finite element models of the medial and lateral plantar longitudinal arch, and to investigate bone and muscle stresses resulting from plantar fasciotomy and major plantar ligament injuries. METHODS: Nonlinear finite element models of the second ray and the fifth ray of plantar longitudinal arches were constructed on the basis of CT and MR images of Virtual Chinese Human "female No. 1". The models assumed a balanced standing load configuration. Three different degrees of passive intrinsic muscle tensions (weak, moderate, or severe) were used in conjunction with simulations of plantar fasciotomy and major plantar ligament injury. FINDINGS: Plantar fasciotomy caused von Mises stress increases in the bones and plantar ligaments while major plantar ligament injuries caused stress increases in the bones, flexor tendons, and plantar fascia. Increasing intrinsic muscle passive tensions decreased stress/strain levels in the medial and lateral arch, and adjusted abnormal tension/compression stress flows of both arches to close to the normal biomechanical states. INTERPRETATION: This study shows that plantar longitudinal arches are concordant combination of bony structures, intrinsic muscles, plantar fascia and ligaments. After plantar ligamentous structure failures, intrinsic muscles have to contribute to stabilize the plantar arches. This mechanism may reduce the risk of developing stress fractures, tendonitis and pain syndrome. PMID- 17118501 TI - Circulating microparticles: pathophysiology and clinical implications. AB - Microparticles (MP) derived from vascular endothelium or circulating blood cells circulate in the peripheral blood. They originate from blebbing and shedding from cell membrane surfaces in physiological and pathological conditions and are present in low concentrations in normal plasma. Increased levels are generated by a number of mechanisms including platelet activation, direct vascular endothelial damage, thrombin activity on the cell surface, C5b-9 activation, and PF4-heparin antibody interaction. Several techniques are currently used to study the generation and nature of circulating microparticles. In particular, the genesis and role of microparticles, derived from platelets, endothelial cells and monocytes, in sepsis (especially meningococcal-induced), heparin-induced thrombocytopenia (HIT), thrombotic thrombocytopenic purpura (TTP), aplastic anaemia, paroxysmal nocturnal haemoglobinuria (PNH) and sickle cell disease (SCD) have been well studied, and provide important insights into the underlying diseases. A defect in the ability to form microparticles leads to the severe bleeding disorder of Scott syndrome, which in turn provides a revealing insight into the physiology of coagulation. In addition the complex role of microparticles in vascular and cardiovascular diseases is an area of immense interest, that promises to yield important advances into diagnosis and therapy. PMID- 17118502 TI - Stroke and cognition: what's hot. PMID- 17118503 TI - Spatial limits on referred touch to an alien limb may reflect boundaries of visuo tactile peripersonal space surrounding the hand. AB - In this study, the spatial limits of referred touch to a rubber hand were investigated. Participants rated the strength of the perceived illusion when the rubber hand was placed in one of six different spatial positions (at a distance of 17.5-67.5cm horizontal from the participant's own hand). The results revealed a significant nonlinear relationship in the strength of the illusion, with the strongest ratings given when the two hands were closest; decaying significantly after a distance of 30cm. The time taken to elicit the illusion followed a similar trend. These results may reflect the response properties of bimodal visuo tactile cells encoding peripersonal space around the hand. PMID- 17118504 TI - The attentional blink reveals sluggish attentional shifting in adolescents with specific language impairment. AB - Rapid processing deficits have been the subject of much debate in the literature on specific language impairment (SLI). Hari and Renvall (2001) [Hari, R. & Renvall, H. (2001). Impaired processing of rapid stimulus sequences in dyslexia. Trends in cognitive sciences, 5, 525-532.] proposed that the source of this deficit can be attributed to sluggish attentional shifting abilities. That is, more time is required to shift attention between stimuli. To test this claim, 26 adolescents with SLI (divided into two subgroups to control for differences in non-verbal intelligence) and 14 controls were presented with a rapid serial visual presentation task. In this task participants were asked to detect two visual targets presented serially with distracter items with varying inter-target intervals (i.e., time difference between targets). This task was designed to elicit an attentional blink (AB). The AB describes the phenomenon whereby non impaired individuals are less likely to report the second of two targets presented within 200-500ms of each other. After controlling for group differences in non-verbal intelligence, the SLI group was found to be significantly less accurate than the control group at successfully reporting the second target at inter-target intervals of 100, 200, 300, 400 and 800ms. The results were interpreted to suggest that adolescents with language impairments have an AB which differs from non-impaired individuals in both magnitude and duration. PMID- 17118505 TI - Biomarkers for oxidative stress status of DNA, lipids, and proteins in vitro and in vivo cancer research. AB - Oxidation and the production of free radicals are an integral part of human metabolism, and oxidative stress is related to many diseases, including cancer and heart disease. The use of biomarkers for oxidative stress may provide further evidence of a causal relationship between oxidative damage to macromolecules (DNA, lipids, and proteins) and cancer. A wide variety of functional assays, both in vivo and ex vivo, include various measures of DNA oxidation (oxidized DNA bases such as 8-OHdG, autoantibodies to oxidized DNA, modified comet assay), lipid oxidation (thiobarbituric acid-reactive substances, exhaled pentane/ethane, low-density lipoprotein resistance to oxidation, isoprostanes), and protein oxidation (protein carbonyls). The objective of this review is to discuss characteristics and methodologic issues for studies involving biomarkers of exposure to antioxidant nutrients and of oxidative stress status. This paper provides an overview on the current knowledge of oxidative DNA, lipid, and protein damage and cancer incidence. PMID- 17118506 TI - Influence of pH and oxygen-inhibited layer on fluoride release properties of fluoride sealant. AB - OBJECTIVES: This study tested the hypothesis that the oxygen-inhibited layer on a light-cured methacrylate based resin and the pH of the storage medium would increase significantly the initial fluoride release and long-term release rate from fluoride dental sealant. METHODS: Forty-eight discs (16-mm diameter x 1-mm thick) were made from FluroShield (<5 wt% NaF) and Helioseal F (<30 wt% fluorosilicate glass) sealants. For each sealant, 24 discs were cured through a Mylar strip that covered the surface and the remaining 24 discs were cured in air allowing formation of the oxygen-inhibited surface. Each specimen in the 24-disc groups was stored individually in 25-mL vials, and divided into four six-vial groups to receive 10 mL of pH4-pH7 (designation of pH 4-7) lactate buffer solutions. The buffer solutions were replaced periodically up to 121 days. The cumulative fluoride release over time was used to determine the coefficients for short-term and long-term release. RESULTS: Two-way ANOVA showed that the mean coefficient values for either sealant were significantly influenced by the curing condition (p<0.0001) and pH (p<0.0001), except for short-term release from NaF sealant. The duration of short-term release was much longer for the fluorosilicate glass sealant. CONCLUSIONS: Both pH and the source of fluoride source incorporated in the sealant play significant roles in fluoride release. PMID- 17118507 TI - Contamination of a toothbrush with antibacterial properties by oral microorganisms. AB - OBJECTIVES: The aim of this study was to examine the contamination and the survival rate of periodontopathic and cariogenic species on new toothbrushes with antibacterial properties (coated bristles with triclosan), after a single use in periodontitis patients. The decontamination effect of the use of toothpaste was also evaluated. METHODS: Ten patients, who consulted the Department of Periodontology, for treatment of chronic periodontitis, were selected. In each patient four different toothbrushes were used. Two quadrants, randomly selected, were each brushed using a different antibacterial toothbrush. In one of these two quadrants toothpaste was used. The same happened with the remaining quadrants, only with regular toothbrushes. After brushing, the toothbrushes were rinsed and stored in room temperature and a dry environment. After 0, 4 and 24h, four tufts, from each toothbrush, were cut and processed for selective and non-selective culturing techniques, followed by identification and quantification of all species found. RESULTS: Immediately after brushing the toothbrushes harbored a significant number of microorganisms, with no statistically significant difference between the two types of brushes (regular and antibacterial). The reduction of microorganisms from 0 to 4h after brushing was statistically significant (p<0.05). The difference was less obvious from 4 to 24h. When toothpaste was used, brushes harbored significantly (p<0.05) lower numbers of colony-forming units (CFU) compared to those without the use of toothpaste. CONCLUSIONS: The antibacterial toothbrush with triclosan coated tufts failed to limit the bacterial contamination. The toothpaste, on the other hand, significantly reduced the contamination of toothbrushes. PMID- 17118508 TI - Pre-hospital pleural decompression and chest tube placement after blunt trauma: A systematic review. AB - Pre-hospital insertion of chest tubes or decompression of air within the pleural space is one of the controversial topics in emergency medical care of trauma patients. While a wide variety of opinions exist medical personnel on the scene require guidance in situations when tension pneumothorax or progressive pneumothorax is suspected. To ensure evidence based decisions we performed a systematic review of the current literature with respect to the diagnostic accuracy in the pre-hospital setting to identify patients with (tension) pneumothorax, the efficacy and safety of performing pleural decompression in the field and the choice of method and technique for the procedure. The evidence found is presented and discussed and recommendations are drawn from the authors' perspective. PMID- 17118509 TI - Epidemiology, trends, and outcome of out-of-hospital cardiac arrest of non cardiac origin. AB - INTRODUCTION: The majority of victims who experience out-of-hospital cardiac arrest (OHCA) have ventricular fibrillation (VF) as the presenting rhythm and are thought to have a cardiac etiology for their arrest. Over the past decade, the incidence of VF OHCA has declined. The aims of this study were to describe the epidemiology of OHCA of non-cardiac origin in Olmsted County MN and to determine the trends that have occurred over time. METHODS: All residents with a traumatic OHCA from 1995 to 2005 were included for analysis. OHCA data were collected prospectively according to the Utstein method. Cardiac arrests were classified as cardiac or non-cardiac in origin and the etiology determined based on autopsy reports, electronic medical records, and/or emergency medical services reports. RESULTS: During the study period, 414 OHCAs were identified, 90 (21.7%) of which were classified as non-cardiac. Mean age was 61.5+/-19.7 years. Response time was 7.73+/-2.9 min, and 40 (44.4%) were bystander-witnessed. Sixty-eight (75.6%) arrests occurred at home, 13 (14.4%) in a public place, and 9 (10%) in other locations. Bystander CPR was performed in 17 (18.9%) cases. The presenting rhythm was VF in 2 (2.2%) cases, PEA in 54 (60%), and asystole in 34 (37.8%). Eight (8.9%) patients survived to hospital discharge. Respiratory failure (35.6%), unknown (15.6%), and pulmonary embolism (13.3%) were the most common etiologies. The mean percentage of arrests due to a non-cardiac cause in three sequential time-periods (1995-1999, 2000-2002, 2003-2005) was 9.4%, 20.1% and 37.7%, respectively. CONCLUSIONS: Over the study period, 21.7% of OHCAs were non-cardiac in origin. PEA was the most common presenting rhythm and respiratory failure the most common etiology. 8.9% of patients survived. The decreasing number of VF arrests may be a contributing factor to the increasing proportion of OHCAs of non cardiac etiology observed in the out-of-hospital setting. PMID- 17118510 TI - Robert Woods (1865-1938): The rationale for mouth-to-mouth respiration. PMID- 17118511 TI - beta-Adrenergic blockade during systemic inflammation: impact on cellular immune functions and survival in a murine model of sepsis. AB - AIM OF THE STUDY: Adrenergic immuno-modulation mediated by beta-adrenergic receptors has been demonstrated. Pharmacological blockade of beta-adrenergic receptors is a therapeutic intervention frequently used in critically ill patients. The effect of beta-adrenergic blockade on cellular immune functions in a critical illness, such as polymicrobial sepsis, has not been investigated. METHODS: Male NMRI-mice were subjected to sham operation or to sepsis (caecal ligation and puncture, CLP) following administration of either the non-selective beta-adrenergic antagonist propranolol (0.5 mg/kg s.c. every 12 h in 1 ml vehicle) or saline 0.9% (1 ml s.c. every 12 h). Mice were kept in metabolic cages and were sacrificed 48 h after induction of sepsis. Survival rate, clinical situation (body weight and temperature, fluid and food intake, urine output), and immunological variables (splenocyte proliferation, apoptosis, and IFN-gamma and IL-6 release) were determined. RESULTS: Administration of propranolol in septic mice increased the splenocyte apoptosis rate, reduced the proliferative capacity of splenocytes, and modulated cellular cytokine release (IL-6, IFN-gamma). This was paralleled by a higher loss of body weight and temperature, and a decreased urine output. Furthermore, treatment with propranolol increased the sepsis induced lethality from 47% up to 68%, respectively. CONCLUSION: beta-Adrenergic blockade was accompanied by alterations of cellular immune functions, a deterioration in the clinical situation and a reduced survival in a murine model of sepsis. These data demonstrate the potential immuno-modulatory effects of beta adrenergic antagonists. PMID- 17118512 TI - Characteristics of volunteers responding to emergencies in the Public Access Defibrillation Trial. AB - OBJECTIVES: To evaluate the characteristics of volunteers responding to emergencies in the North American Public Access Defibrillation (PAD) Trial. METHODS: The PAD Trial was a prospective evaluation of cardiac arrest survival in community facilities randomized to cardiopulmonary resuscitation (CPR) or to CPR with automated external defibrillators (AEDs). The PAD volunteers' characteristics were analyzed using Poisson regression clustered on the facility and offset by the number of emergency episodes to which volunteers were exposed. RESULTS: A total of 19,320 volunteers in 1260 facilities were trained to provide emergency care. Of these, 8169 volunteers were participating actively at their facility during a time when one or more emergency episodes occurred. There were 1971 emergency episodes responded to by 1245 volunteers. The treatment arm (CPR only versus CPR+AED) was not associated with the likelihood of volunteer participation in an episode. Likewise, the volunteers' age or sex did not affect response. Volunteers more likely to respond were supervisory/management or security personnel, non-minority participants, volunteers with previous CPR training, volunteers with previous experience in emergency care and those who passed the PAD CPR skills follow-up test. Volunteers who had a formal education beyond a high school level were less likely to respond. CONCLUSIONS: Volunteers with previous emergency training and positions of responsibility in their facility had a greater likelihood of participation in medical emergencies in the PAD Trial. PMID- 17118514 TI - Effective ventilation following emergency needle cricothyroidotomy. PMID- 17118515 TI - Interpersonal defensiveness and diminished perceptual acuity for the odor of a putative pheromone: androstenone. AB - This study assessed the relationship of repressive coping and defensiveness to the perception of androstenone, which is a putative human pheromone with relevance to social perception. In Experiment 1, 34 men and 34 women between the ages of 16-28 sniffed pairs of bottles containing silicone (the solvent control) paired with either isoamyl acetate (IAA) or androstenone in an eight trial, two alternative forced-choice task. Participants chose which hand they believed the odor was in, and rated the odor's intensity and their confidence in their response on a 0-10 scale. Defensiveness was measured with the Eysenck Personality Questionnaire (EPQ) L scale. As predicted, defensiveness was associated with decreased perception of androstenone, but not IAA. Decreased detection accuracies, confidence, and intensity ratings for androstenone were associated with high defensiveness. The effect was stronger among women. None of the IAA detection variables correlated with the L scale for either women or men. In Experiment 2, 22 women and 18 men between the ages of 18 and 27 were given 4 concentrations of IAA ranging from subthreshold to suprathreshold, and a blank blank control. Detection accuracies, confidence ratings, and intensity ratings for IAA were not related to defensiveness for any of the concentrations, for either men or for women. The results are discussed in terms of motives to seek social approval and avoid social disapproval as they may relate to diminished awareness of androstenone. PMID- 17118516 TI - Domestic violence and vagal reactivity to peer provocation. AB - This paper examined whether individual differences in children's vagal reactivity to peer provocation were related to domestic violence within the family. It also examined the question of whether conduct-problem children who show vagal augmentation to peer provocation come from families with high levels of domestic violence. During the peer provocation, children were expecting to interact with a difficult peer while vagal reactivity was assessed. Groups were divided into children who showed vagal augmentation and vagal suppression to the stressful peer interaction. Findings indicated that conduct-problem children who showed vagal augmentation to interpersonal challenge came from families with the highest levels of domestic violence. Vagal augmentation was also associated with a greater number of conduct-related problems for those children exposed to high levels of domestic violence. Discussion highlights the role of individual differences in physiological reactivity in understanding children's behavior problems in relation to domestic violence. PMID- 17118517 TI - Effects of prepulse intensity, duration, and bandwidth on perceived intensity of startling acoustic stimuli. AB - Intense abrupt stimuli can elicit a startle reflex; a weak "prepulse" 30-300 ms earlier can reduce both startle and perceived stimulus intensity. Prepulse inhibition (PPI) of startle, an operational measure of sensorimotor gating, is used to understand brain disorders characterized by gating deficits. Compared to startle, PPI of perceived stimulus intensity (PPIPSI) may provide information that is distinct, and easier to acquire and analyze. To develop this experimental measure, we examined PPIPSI under different stimulus conditions. Both PPI and PPIPSI exhibited a non-linear relationship to prepulse intensity, with prepulses 15 dB(A) above background causing maximal inhibition of both measures. A 50 ms broadband noise prepulse produced maximal PPI and PPIPSI, whereas 5 and 20 ms pure tone prepulses produced maximal PPIPSI and PPI, respectively. PPIPSI is a robust, parametrically sensitive and "low tech" measure of sensory gating that may become a valuable tool for understanding the biology of certain mental disorders. PMID- 17118518 TI - Musically induced arousal affects pain perception in females but not in males: a psychophysiological examination. AB - The present study investigated affective and physiological responses to changes of tempo and mode in classical music and their effects on heat pain perception. Thirty-eight healthy non-musicians (17 female) listened to sequences of 24 music stimuli which were variations of 4 pieces of classical music. Tempo (46, 60, and 95 beats/min) and mode (major and minor) were manipulated digitally, all other musical elements were held constant. Participants rated valence, arousal, happiness and sadness of the musical stimuli as well as the intensity and the unpleasantness of heat pain stimuli which were applied during music listening. Heart rate, respiratory rate and end-tidal PCO(2) were recorded. Pain ratings were highest for the fastest tempo. Also, participants' arousal ratings, their respiratory rate and heart rate were accelerated by the fastest tempo. The modulation of pain perception by the tempo of music seems to be mediated by the listener's arousal. PMID- 17118519 TI - Reconstructive plastic surgery in the treatment of vulvar carcinomas. AB - OBJECTIVE: The results obtained using plastic surgery reconstruction in 207 patients with a primary or recurrent vulvar carcinoma were analyzed with regard to the surgical procedures applied, pre-treatment and post-operative findings, along with the long-term oncological disease course. STUDY DESIGN: Standardized data concerning the surgical procedures applied and clinical factors were collected in a databank and statistically analyzed. RESULTS: The flaps employed were termed either 'local' (cutaneous or fasciocutaneous; n=84) or 'regional' (myocutaneous, n=123). For local flaps, the rate of secondary healing was 31%, dropping to 20% for regional flaps. Such healing disturbances often affected the donor region and did not lead to lasting clinical problems such as stenosis or distortion. Severe disturbances of wound healing (loss of more than 10%) were not observed in local flaps, whereas such problems were encountered in 5.9% of regional flaps. Gluteal thigh flaps were most frequently applied and were also the most successful type of myocutaneous reconstruction. Therapy was selected on an individual basis according to tumor status. CONCLUSION: Plastic surgery reconstruction broadens the range of operative therapies available for the treatment of vulvar carcinomas, especially those at an advanced stage or recurrent tumors, and leads to a favorable oncological disease course as well as improved cosmetic results. PMID- 17118520 TI - Pregnancy outcome in primiparae of advanced maternal age. AB - OBJECTIVE: To investigate the impact of maternal age on singleton pregnancy outcome, taking into account intermediate and confounding factors. STUDY DESIGN: In this population-based retrospective cohort study, perinatal data of primiparous women aged 35 years or more (n = 2970), giving birth to a singleton child of at least 500 g, were compared to data of primiparous women aged 25-29 years old (n = 23,921). Univariate analysis was used to assess the effect of maternal age on pregnancy outcomes. The effects of intermediate (hypertension, diabetes and assisted conception) and confounding factors (level of education) were assessed through multivariable logistic regression analysis. RESULTS: Older maternal age correlated, independently of confounding and intermediate factors, with very preterm birth (gestational age <32 weeks) [adjusted odds ratio (AOR) 1.51, 95% confidence intervals (CI) 1.04-2.19], low birth weight (birth weight <2500 g) (AOR 1.69, 95% CI 1.47-1.94) and perinatal death (AOR 1.68, 95% CI 1.06 2.65). CONCLUSION: Maternal age is an important and independent risk factor for adverse pregnancy outcome. PMID- 17118521 TI - The mammographic appearance of breast carcinomas of invasive ductal type: relationship with clinicopathological parameters, biological features and prognosis. AB - OBJECTIVES: To evaluate the clinical significance of the mammographic appearance of tumors in 411 patients with infiltrating ductal carcinoma of the breast. STUDY DESIGN: Tumors were classified into five radiographic subgroups: spiculated mass (A-type), diffuse changes with or without suspicious microcalcifications (B type), microcalcifications with a mass (C-type), circumscribed (D-type), and not visible (E-type). Intratumoral levels of estrogen (ER) and progesterone (PR) receptors, c-erbB-2, EGFR, pS2, cathepsin D and tPA, ploidy and S-phase fraction, were analysed in a significant number of cases. RESULTS: A-type A radiographic pattern was detected in 234 patients (57%), B-type in 46 (11%), C-type in 46 (11%), D-type in 68 (17%), and E-type in 17 patients (4%). On the other hand, a total of 155 tumors (37.8%) showed microcalcifications. The percentage of tumors showing A-type pattern was more frequent in postmenopausal women, in well differentiated tumors, and in those showing higher levels of ER, pS2 of tPA. However, B-type pattern was detected in a high percentage of premenopausal women and in those showing larger tumors, positive nodes, poor differentiation or high S-phase fraction. Cox multivariate analysis showed that B-type pattern and the absence of microcalcifications were factors significantly associated to high risk for relapse. CONCLUSIONS: Our results suggest that the mammographic appearance of tumor may to provide useful clinical information in addition to classical prognostic factor in infiltrating ductal carcinoma of the breast. PMID- 17118522 TI - Distinguishing benign and malignant pelvic masses: the value of different diagnostic methods in everyday clinical practice. AB - OBJECTIVE: To optimize referral to specialized gynaecologists for surgical treatment of ovarian cancer by improving preoperative discrimination between benign and malignant pelvic tumours. STUDY DESIGN: In a prospective multicentre study 143 patients with a pelvic mass were included. At several occasions during the diagnostic work-up the gynaecologist estimated the chance of malignancy (educated guess/expert opinion). MRI in the local setting was suggested for uncertain cases. All MRI images were reviewed by an expert radiologist. The datasheet designed for the study further allowed for determining the risk of malignancy index (RMI). RESULTS: The diagnostic accuracy of the gynaecologist's final estimation of the chance of malignancy and the calculated RMI were comparable (area under the ROC curve of 0.87 and 0.86). MRI did not improve the accuracy of the diagnostic work-up for the study population as a whole. Subgroup analysis did however show improved diagnostic accuracy in cases with an estimated chance of malignancy between 20 and 80% when the MRI was read by an expert radiologist. CONCLUSION: Patient selection for surgery of a pelvic mass should be based on the chance of malignancy as assigned by the referring gynaecologists. In case of uncertainty MRI improves diagnostic accuracy, when judged by an expert. PMID- 17118523 TI - Residue cluster additivity of thermodynamic stability in the hydrophobic core of mesophile vs. hyperthermophile rubredoxins. AB - The branched sidechain residues 24 and 33 in the hydrophobic core of rubredoxin differ between the Clostridium pasteurianum (Cp) and Pyrococcus furiosus (Pf) sequences. Their X-ray structures indicate that these two sidechains are in van der Waals contact with each other, while neither appears to significantly interact with the other nonconserved residues. The simultaneous interchange of residues 24 and 33 between the Cp and Pf rubredoxin sequences yield a complementary pair of hybrid proteins for which the sum of their thermodynamic stabilities equals that of the parental rubredoxins. The 1.2 kcal/mol change arising from this two residues interchange accounts for 21% of the differential thermodynamic stability between the mesophile and hyperthermophile proteins. The additional interchange of the sole nonconserved aromatic residue in the hydrophobic core yields a 0.78 kcal/mol deviation from thermodynamic additivity. PMID- 17118524 TI - The structure at 2 A resolution of Phycocyanin from Gracilaria chilensis and the energy transfer network in a PC-PC complex. AB - Phycocyanin is a phycobiliprotein involved in light harvesting and conduction of light to the reaction centers in cyanobacteria and red algae. The structure of C phycocyanin from Gracilaria chilensis was solved by X-ray crystallography at 2.0 A resolution in space group P2(1). An interaction model between two PC heterohexamers was built, followed by molecular dynamic refinement. The best model showed an inter-hexamer rotation of 23 degrees . The coordinates of a PC heterohexamer (alphabeta)(6) and of the PC-PC complex were used to perform energy transfer calculations between chromophores pairs using the fluorescence resonance energy transfer approach (FRET). Two main intra PC ((I)beta(3)(82)- >(I)alpha(1)(84)-->(I)alpha(5)(84)-->(I)beta(6)(82) and (I)beta(3)(153)- >(I)beta(5)(153)) and two main inter PC ((I)beta(6)(82)-->(II)beta(3)(82) and (I)beta(5)(153)-->(II)beta(3)(153)) pathways were proposed based on the values of the energy transfer constants calculated for all the chromophore pairs in the hexamer and in the complex. PMID- 17118525 TI - What is driving the EU Burden-sharing Agreement: efficiency or equity? AB - Under the Kyoto Protocol the European Union agreed to reduce emissions of greenhouse gases by 8 percent. The Burden-Sharing Agreement (BSA) redistributes the reduction target among the member states. The purpose of this paper is to evaluate the BSA. To determine if cost efficiency was considered, marginal abatement costs (MACs) are first calculated based on an estimation of the directional output distance function using country production data for 1990-2000. MACs, together with equity indicators, are then regressed on the emission change targets. The main conclusion is that both efficiency and equity were important aspects considered in the settlement. PMID- 17118526 TI - Importance of information on tillage practices in the modelling of environmental processes and in the use of environmental indicators. AB - Tillage has been and will always be integral to crop production. Tillage can result in the degradation of soil, water, and air quality. Of all farm management practices, tillage may have the greatest impact on the environment. A wide variety of tillage equipment, practices and systems are available to farmers, providing opportunities to enhance environmental performance. These opportunities have made tillage a popular focus of environmental policies and programs such as environmental indicators for agriculture. This paper provides a very brief examination of the role of tillage in crop production, its effect on biophysical processes and, therefore, its impact on the environment. Models of biophysical processes are briefly examined to demonstrate the importance of tillage relative to other farm management practices and to demonstrate the detail of tillage data that these models can demand. The focus of this paper is an examination of the use of information on tillage in Canada's agri-environmental indicators initiative, National Agri-environmental Health Analysis and Reporting Program (NAHARP). Information on tillage is required for several of the indicators in NAHARP. The type of data used, its source, and its quality are discussed. Recommendations regarding the collection of tillage data and use of tillage information are presented. PMID- 17118527 TI - Oxidation and detoxification of pentachlorophenol in aqueous phase by ozonation. AB - The degradation and detoxification performance of ozonation in treating pentachlorophenol (PCP) contaminated wastewater was determined. All experiments were conducted in a bench scale glass column equipped with ceramic diffuser and a lab-scale ozone generator under ambient temperature and pH 7. The decomposition rate of PCP in this study was primarily controlled by the ozone mass transfer rate from gas to liquid phases. Principal intermediates found were 2,3,4,6- and 2,3,5,6-tetrachlorophenols (TeCP) and phenol. PCP seems to be more vulnerable to ozone than its intermediates. A bioluminescence technique was used to evaluate the toxicity of PCP with Vibrio fisheri NRRL B-11177 as the test bacterium, and the EC(50) of PCP was found to be 1.0 mg l(-1). Detoxification occurred as the PCP and TeCP reacted with ozone and decomposed to less chlorinated congeners and phenol. PMID- 17118528 TI - Analysis of gene regulatory network models with graded and binary transcriptional responses. AB - The steep sigmoid framework developed by Plahte and Kjoglum [Plahte, E., Kjoglum, S., 2005. Analysis and generic properties of gene regulatory networks with graded response functions. Phys. D 201, 150-176, doi:10.1016/j.physd.2004.11.014] provides a uniform description of gene regulatory networks in which there may be both graded and binary transcriptional responses, as well as a method for analysing the models developed. Here we extend this framework. We show that there is a relation between the location of steady states and the feedback structure of a system, thus generalising existing results for Boolean type models. In addition, we justify underlying assumptions and generic features of the modelling framework in terms of biology and generalise the overall approach to take into account that each transcription factor only regulates one gene at a given threshold. By this assumption, the analysis of the models are greatly simplified. PMID- 17118529 TI - A physiological role for glucuronidated thyroid hormones: preferential uptake by H9c2(2-1) myotubes. AB - Conjugation reactions are important pathways in the peripheral metabolism of thyroid hormones. Rat cardiac fibroblasts produce and secrete glucuronidated thyroxine (T4G) and 3,3',5-triiodothyronine (T3G). We here show that, compared to fibroblasts from other anatomical locations, the capacity of cardiofibroblasts to secrete T4G and T3G is highest. H9c2(2-1) myotubes, a model system for cardiomyocytes, take up T4G and T3G at a rate that is 10-15 times higher than that for the unconjugated thyroid hormones. T3 and T4, and their glucuronides, stimulate H9c2(2-1) myoblast-to-myotube differentiation. A substantial beta glucuronidase activity was measured in H9c2(2-1) myotubes, and this confers a deconjugating capacity to these cells, via which native thyroid hormones can be regenerated from glucuronidated precursors. This indicates that the stimulatory effects on myoblast differentiation are exerted by the native hormones. We suggest that glucuronidation represents a mechanism to uncouple local thyroid hormone action in the heart from that in other peripheral tissues and in the systemic circulation. This could represent a mechanism for the local fine-tuning of cardiac thyroid hormone action. PMID- 17118530 TI - Zinc-transporter genes in human visceral and subcutaneous adipocytes: lean versus obese. AB - Zinc ions influence adipose tissue metabolism by regulating leptin secretion and by promoting free fatty acid release and glucose uptake. The mechanisms controlling zinc metabolism in adipose tissue are unknown. We therefore examined the gene-expression levels of a number of zinc-transporting proteins in adipose tissue, comparing subcutaneous fat with visceral fat from lean and obese humans. Both ZnT-proteins responsible for zinc transport from cytosol to extracellular compartments and intracellular vesicles and Zip-proteins responsible for zinc transport to the cytoplasm were expressed in all samples. This suggests that zinc metabolism in adipocytes is actively controlled by zinc-transporters. The expression levels were different in lean and obese subjects suggesting a role for these proteins in obesity. Furthermore, the expression levels were different from subcutaneous fat to intra-abdominal fat suggesting that the metabolic activity in adipocytes is to some extent dependent upon zinc and the activity of zinc transporting proteins or vice versa. PMID- 17118531 TI - Interferon causes no myasthenia in a seropositive patient with multiple sclerosis. AB - We describe a successful outcome of long-term interferon beta-1b therapy in a patient who had multiple sclerosis (MS) with positive serum autoantibody to muscle acetylcholine receptor (AChR-Ab). Because of the reported possible causative linkage between interferon beta-1b and myasthenia gravis (MG), the presence of the pathogenic antibody complicated therapeutic strategies. We carefully observed the patient for further 6 months before the treatment, excluding symptomatic MG. The interferon beta-1b therapy then provided a clinical benefit. Hopefully this report will allow MS patients in similar situations to make more rapid, unprejudiced judgments than our patients. PMID- 17118532 TI - Characteristics and mobility of heavy metals in an MSW landfill: implications in risk assessment and reclamation. AB - To assess the reclamation feasibility of a landfill, the characteristics, distribution, and mobility of heavy metals in the landfill were investigated. The refuse was characterized as containing high concentrations of heavy metals, a relatively high pH, and a high ratio of NH(4)-N to total nitrogen (TN). The results of heavy metal distribution showed that relatively high levels of heavy metals were accumulated in the landfill. Sequential extraction revealed that the relative amounts of heavy metals were different in the samples. Zn demonstrated the greatest mobility compared to other heavy metals, whereas Cd was well retained in the landfill. Leaching experiments indicated that the mobility of heavy metals in the landfill was generally low under normal neutral conditions. However, release of heavy metals can be increased greatly when medium conditions become more acidic and aerobic. PMID- 17118533 TI - Study of ambient air particulates pollutants near Taichung airport sampling site in central Taiwan. AB - This study monitored the atmospheric pollutants at Taichung airport (TA) sampling site during September to December of 2005 near Taiwan Strait in central Taiwan. The distributions of the particle concentrations between the TSP, coarse and fine were also displayed at TA sampling site. Analysis the average percentage of various kinds of metallic elements, the data obtained here indicated that the average percentage concentrations of metallic elements Fe and Mg were higher in fine particulate than that of the coarse particulate at Taichung airport sampling site. These foundry factories were by the TA sampling site was the possible reason responsible for this result. In addition, by a non-parametric (Spearman) correlation analysis, the results indicated that Fe, Mn and Pb have high correlation coefficients of metallic elements in different particle size. The results indicate that the airport pollutants might come from similar pollutant sources at this airport sampling site. In addition, high correlation coefficients of non-airport pollutants were observed on the ionic species of SO(4)(2-), NH(4)(+) and K(+). The r(sp) correlate values between closed to r(sp)=0.7 and 0.9 at different particles size mode. Besides, the ionic Ca(2+) has high negative values (r(sp)=-0.66, -0.66 and 0.61) with the increasing of the temperature, relative humidity and wind speed, respectively at fine particle size. PMID- 17118534 TI - Absorption of carbon dioxide by raw and treated dye-bath effluents. AB - A bench-scale experimental apparatus, consisting of a glass column randomly packed with cylindrical glass rings, was utilized for the removal of carbon dioxide (CO(2)) by chemical absorption. Various liquid solvents were used, in a batch mode, circulating between the column and a reservoir vessel, and contacting the gas in counter current flow. The absorptive capacity of the liquid solvents and the absorption kinetics were studied by obtaining the respective 'breakthrough curves'. Aqueous solutions of sodium hydroxide (NaOH) were initially utilized in order to demonstrate the experimental apparatus function. Dye-bath effluent proved highly effective due to its high alkalinity and the capacity of the hydrolyzed "azo-reactive" dyes to react with CO(2). Decolorized dye-bath effluent (treated with FeSO(4)) can also be used as a chemical solvent for the absorption of CO(2); however, its absorptive capacity is much lower compared to the raw wastewater. The above technique is also a very effective method to neutralize industrial effluents. PMID- 17118535 TI - Use of natural clinoptilolite for the removal of lead, copper and zinc in fixed bed column. AB - This work deals with the removal of lead, copper and zinc from aqueous solutions by using natural zeolite (clinoptilolite). Fixed bed experiments were performed, using three different volumetric flow rates of 5, 7 and 10bed volume/h, under a total normality of 0.01N, at initial pH of 4 and ambient temperature (25 degrees C). The removal efficiency increased when decreasing the flow rate and the following selectivity series was found: Pb(2+)>Zn(2+)> or =Cu(2+). Conductivity measurements showed that lead removal follows mainly ion exchange mechanism, while copper and zinc removal follows ion exchange and sorption mechanism as well. PMID- 17118536 TI - The effect of isosaccharinic acid (ISA) on the mobilization of metals in municipal solid waste incineration (MSWI) dry scrubber residue. AB - Co-landfilling of incineration ash and cellulose might facilitate the alkaline degradation of cellulose. A major degradation product is isosaccharinic acid (ISA), a complexing agent for metals. The impact of ISA on the mobility of Pb, Zn, Cr, Cu and Cd from a municipal solid waste incineration dry scrubber residue was studied at laboratory using a reduced 2(5-1) factorial design. Factors investigated were the amount of calcium isosaccharinate (Ca(ISA)(2)), L/S ratio, temperature, contact time and type of atmosphere (N(2), air, O(2)). The effects of pH and Ca(ISA)(2) as well as other factors on the leaching of metals were quantified and modelled using multiple linear regression (alpha=0.05). Cd was excluded from the study since the concentrations were below the detection limit. The presence of Ca(ISA)(2) resulted in a higher leaching of Cu indicating complex formation. Ca(ISA)(2) alone had no effect on the leaching of Pb, Zn and Cr. A secondary effect on the mobilization was predicted to occur since Ca(ISA)(2) had a positive effect on the pH and the leaching of Pb, Zn and Cr increased with increasing pH. The leaching of Pb varied from 24 up to 66 wt.% of the total Pb amount (1.74+/-0.02 g(kgTS)(-1)) in the dry scrubber residue. The corresponding interval for Zn (7.29+/-0.07 g(kgTS)(-1)) and Cu (0.50+/-0.02 g(kgTS)(-1)) were 0.5-14 wt.% of Zn and 0.8-70wt.% of Cu. Maximum leaching of Cr (0.23+/-0.03 g(kgTS)(-1)) was 4.0 wt.%. At conditions similar to a compacted and covered landfill (4 degrees C, 7 days, 0 vol.% O(2)) the presence of ISA can increase the leaching of Cu from 2 to 46 wt.% if the amount of cellulose-based waste increases 20 times, from the ratio 1:100 to 1:5. As well, the leaching of Pb, Zn, and Cr can increase from 32 to 54 wt.% (Pb), 0.8-8.0 wt.% (Zn), and 0.5 to 4.0 wt.% (Cr) depending on the amount of cellulose and L/S ratio and pH value. Therefore, a risk (alpha=0.05) exists that higher amounts of metals are leached from landfills where cellulose-containing waste and ash are co-disposed. This corresponds to an additional 29 t of Pb and 17 t of Cu leached annually from a compacted and covered landfill in the north of Sweden. PMID- 17118537 TI - Photocatalytic performance of Sn-doped TiO2 nanostructured mono and double layer thin films for Malachite Green dye degradation under UV and vis-lights. AB - Nanostructure Sn(4+)-doped TiO(2) based mono and double layer thin films, contain 50% solid ratio of TiO(2) in coating have been prepared on glass surfaces by spin coating technique. Their photocatalytic performances were tested for degradation of Malachite Green dye in solution under UV and vis irradiation. Sn(4+)-doped nano-TiO(2) particle a doping ratio of about 5[Sn(4+)/Ti(OBu(n))(4); mol/mol%] has been synthesized by hydrotermal process at 225 degrees C. The structure, surface and optical properties of the thin films and/or the particles have been investigated by XRD, BET and UV/vis/NIR techniques. The results showed that the double layer coated glass surfaces have a very high photocatalytic performance than the other one under UV and vis lights. The results also proved that the hydrothermally synthesized nano-TiO(2) particles are fully anatase crystalline form and are easily dispersed in water. The results also reveal that the coated surfaces have hydrophilic property. PMID- 17118538 TI - A comparative study of linear and non-linear regression analysis for ammonium exchange by clinoptilolite zeolite. AB - Ammonium ion exchange from aqueous solution using clinoptilolite zeolite was investigated at laboratory scale. Batch experimental studies were conducted to evaluate the effect of various parameters such as pH, zeolite dosage, contact time, initial ammonium concentration and temperature. Freundlich and Langmuir isotherm models and pseudo-second-order model were fitted to experimental data. Linear and non-linear regression methods were compared to determine the best fitting of isotherm and kinetic model to experimental data. The rate limiting mechanism of ammonium uptake by zeolite was determined as chemical exchange. Non linear regression has better performance for analyzing experimental data and Freundlich model was better than Langmuir to represent equilibrium data. PMID- 17118539 TI - Photocatalytic degradation of pollutants from Elcogas IGCC power station effluents. AB - The aim of this work is to improve the quality of water effluents coming from Elcogas IGCC power station (Puertollano, Spain) with the purpose of fulfilling future more demanding normative, using heterogeneous photocatalytic oxidation processes (UV/H(2)O(2)/TiO(2) or ZnO). The efficiency of photocatalytic degradation for the different catalysts (TiO(2) and ZnO) was determined from the analysis of the following parameters: cyanides, formates and ammonia content. In a first stage, the influence of two parameters (initial concentration of H(2)O(2) and amount of catalyst) on the degradation kinetics of cyanides and formates was studied based on a factorial experimental design. pH was always kept in a value >9.5 to avoid gaseous HCN formation. The degradation of cyanides and formates was found to follow pseudo-first order kinetics. Experimental kinetic constants were fitted using neural networks (NNs). The mathematical model reproduces experimental data within 90% of confidence and allows the simulation of the process for any value of parameters in the experimental range studied. Moreover, a measure of the saliency of the input variables was made based upon the connection weights of the neural networks, allowing the analysis of the relative relevance of each variable with respect to the others. Results showed that the photocatalytic process was effective, being the degradation rate of cyanides about five times higher when compared to removal of formates. Finally, the effect of lowering pH on the degradation of formates was evaluated after complete cyanides destruction was reached (10 min of reaction). Under the optimum conditions (pH 5.2, [H(2)O(2)]=40 g/l; [TiO(2)]=2g/l), 100% of cyanides and 92% of initial NH(3) concentration are degraded after 10 min, whereas 35 min are needed to degrade 98% of formates. PMID- 17118540 TI - Equilibrium and kinetics studies for adsorption of direct blue 71 from aqueous solution by wheat shells. AB - This paper deals with the application of wheat shells (WS), an agricultural by product, for the removal of direct blue 71 (DR) from aqueous solution. The characteristics of WS surface, such as surface area, Bohem titration and scanning electron microscopy (SEM) were obtained. The removal of direct blue 71 onto WS from aqueous solution was investigated by using parameters, such as pH, temperature, adsorbent dose, contact time and initial concentration. The adsorption process attains equilibrium within 36 h. The extent of dye removal decreased with increasing adsorbent dosage and also increased with increasing contact time, temperature, in solution concentration. Optimum pH value for dye adsorption was determined between 6 and 8. The experimental data were analysed by the Langmuir and Freundlich models of adsorption. It was found that the Langmuir equation fit better than the Freundlich equation. Maximum adsorption capacity (Q(m)) was calculated as at different temperatures (293, 303 and 313 K) 40.82, 45.66 and 46.30 mgg(-1), respectively. In addition, the adsorption data obtained at different temperatures of DR by WS were applied to pseudo first-order, pseudo second-order and Weber-Morris equations, and the rate constants of first-order adsorption (k(1)), the rate constants of second-order adsorption (k(2)) and intraparticle diffusion rate constants (k(3)) at these temperatures were calculated, respectively. The rates of adsorption were found to conform to pseudo second-order kinetics with good correlation (R(2)>or=0.9904). Also, free energy of adsorption (DeltaG degrees), enthalpy (DeltaH degrees), and entropy (DeltaS degrees) changes were determined to predict the nature of adsorption. Furthermore, the results indicate that WS could be employed as a low-cost alternative to other adsorbents in the removal of direct blue 71 from aqueous solution. PMID- 17118541 TI - A combined electrocoagulation-sorption process applied to mixed industrial wastewater. AB - The removal of organic pollutants from a highly complex industrial wastewater by a aluminium electrocoagulation process coupled with biosorption was evaluated. Under optimal conditions of pH 8 and 45.45 Am(-2) current density, the electrochemical method yields a very effective reduction of all organic pollutants, this reduction was enhanced when the biosorption treatment was applied as a polishing step. Treatment reduced chemical oxygen demand (COD) by 84%, biochemical oxygen demand (BOD(5)) by 78%, color by 97%, turbidity by 98% and fecal coliforms by 99%. The chemical species formed in aqueous solution were determined. The initial and final pollutant levels in the wastewater were monitored using UV-vis spectrometry and cyclic voltammetry. Finally, the morphology and elemental composition of the biosorbent was characterized with scanning electron microscopy (SEM) and energy dispersion spectra (EDS). PMID- 17118542 TI - Production of lightweight aggregates from mining residues, heavy metal sludge, and incinerator fly ash. AB - In this study, artificial lightweight aggregate (LWA) manufactured from recycled resources was investigated. Residues from mining, fly ash from an incinerator and heavy metal sludge from an electronic waste water plant were mixed into raw aggregate pellets and fed into a tunnel kiln to be sintered and finally cooled rapidly. Various feeding and sintering temperatures were employed to examine their impact on the extent of vitrification on the aggregate surface. Microstructural analysis and toxicity characteristic leaching procedure (TCLP) were also performed. The results show that the optimum condition of LWA fabrication is sintering at 1150 degrees C for 15 min with raw aggregate pellets fed at 750 degrees C. The rapidly vitrified surface envelops the gas produced with the increase in internal temperature and cooling by spraying water prevents the aggregates from binding together, thus forming LWA with specific gravity of 0.6. LWA produced by sintering in tunnel kiln shows good vitrified surface, low water absorption rate below 5%, and low cylindrical compressive strength of 4.3 MPa. In addition, only trace amounts of heavy metals were detected, making the LWA non-hazardous for construction use. PMID- 17118543 TI - Adsorption of arsenic from aqueous solution on synthetic hydrous stannic oxide. AB - The hydrated stannic oxide (HSO) was synthesized and arsenic adsorption behaviour is reported. HSO is found to be amorphous, and stable thermally up to 700 degrees C. The adsorption of As(III) is much higher than As(V) in the drinking water pH (6.5-8.5) range. The time required for reaching equilibrium is 4.0 and 3.0 h, respectively for As(III) and As(V). The adsorption kinetic data obtained at pH 7.0 (+/-0.1) and temperature 27(+/-1) degrees C follow the pseudo-second-order kinetic model best (R(2)>0.98). The analyzes of isotherm adsorption data by two parameter isotherm model equations show the order to obey: Langmuir>Freundlich>Temkin for As(III), and Langmuir>Temkin>Freundlich for As(V). The monolayer adsorption capacities (mg/g) obtained for As(III) and As(V) are 15.85 and 4.30, respectively. Excepting phosphate, other anions studied show no adverse effect on adsorption of As(III) onto HSO. A fixed bed HSO packed column (internal diameter 0.70 cm, bed height 3.7 cm and particle size 0.14-0.29 mm) generates 2400 and 450 BV of potable water (As<0.01 mg/L), respectively, for As(III) and As(V) from arsenic spiked (1.0 mg/L) water samples at pH 7.0 (+/ 0.1), which indicated that HSO can be used as an efficient scavenger for As(III) from the contaminated water. PMID- 17118544 TI - Comparison of reductive dechlorination of p-chlorophenol using Fe0 and nanosized Fe0. AB - Chlorophenols, as a kind of important contaminants in groundwater, are toxic and difficult to biodegrade. Laboratory tests were conducted to examine zero-valent iron as an enhancing agent in the dechlorination of chlorinated organic compounds. Nanoscale iron particles were synthesized from common precursors KBH(4) and FeSO(4). Batch experiments were performed to investigate the reduction of p-chlorophenol (4-CP) by both common Fe(0) and nanoscale Fe(0). Comparison of 300 mesh/100 mesh/commercial reductive iron powders showed that size of iron particles played an important role in reduction process. Initial concentration and pretreatment of iron particles also influenced the chlorination rate. Nanoscale Fe(0) offered much more advantages for treatment of 4-CP compared with common iron particles, such as stability and durability. And they can be used to treat contaminants in groundwater over a long time. Among different parts of synthesized nanoscale iron particle solution, the very fine particles were the major agent for treatment of pollutants. As for preservation of nanoscale Fe(0), ethanol was recommended. PMID- 17118545 TI - Amperometric determination of hydrazine at manganese hexacyanoferrate modified graphite-wax composite electrode. AB - Fabrication, characterization and application of a manganese hexacyanoferrate (MnHCF) modified graphite-wax composite electrode are described. The MnHCF mixed with graphite powder was dispersed into molten paraffin wax to yield a conductive composite, which was used as electrode material to construct a renewable three dimensional MnHCF modified electrode. The characterization of the modified electrode has been studied by electrochemical techniques. The cyclic voltammogram of the MnHCF modified graphite-wax composite electrode prepared under optimum composition, showed a well-defined redox couple due to Fe(CN)(6)(4-)/Fe(CN)(6)(3 ) system. The electrocatalytic oxidation of hydrazine by MnHCF modified graphite wax composite electrode has been investigated in an attempt to develop a new sensor for its determination. It was found that the mediator catalyzed the oxidation of hydrazine. The electrocatalytic oxidation of hydrazine was also studied under hydrodynamic and chronoamperometric conditions. The anodic current increases linearly with increase in the concentration of hydrazine in the range of 3.33x10(-5)M to 8.18x10(-3)M. The detection limit was found to be 6.65x10(-6)M (S/N=3). The modified electrode can also be used for on-line detection of hydrazine. The proposed method has also been applied for the determination of hydrazine in photographic developer solution. PMID- 17118546 TI - Influence of Fenton oxidation on soil organic matter and its sorption and desorption of pyrene. AB - The influences of Fenton oxidation on the content and composition of soil organic matter (SOM) and the consequent change of its sorption and desorption of pyrene were investigated using three soil samples. The results showed that both the content and the composition of the SOM changed, with total SOM content decreasing. The content of humic acid (HA) was reduced, while the content of humin did not change significantly, however the content of fulvic acid (FA) had a tendency to increase. Correlation analysis of soil-water distribution coefficient (K(d)) and different parts of the SOM reveals that humin and HA are the key factors controlling the sorption of pyrene. Organic carbon normalized K(d) (K(OC)) varied to different extents after Fenton oxidation due to the change of SOM composition. The reduction of K(OC) is significant in Soils 1 and 2 where large part of HA was reduced to FA, whose sorption ability is low. The change of K(OC) by oxidation in Soil 3 is not so significant due to that the percentage of humin and HA in Soil 3 did not change greatly after oxidation. Desorption was hysteretic in all cases, and humin percentage was found to be the key factor on the extent of desorption hystersis. Oxidation made desorption more hysteretic due to the elevated proportion of humin. PMID- 17118547 TI - Environmental risk assessment of hydrofluoropolyethers (HFPEs). AB - Hydrofluoropolyethers (HFPEs), a new family of linear oligomeric fluorinated fluids, are being used as potential replacements for halon, hydrochlorofluorocarbons (HCFCs) and perfluorocarbons (PFCs) that have been listed as ozone depleting substances and/or greenhouse gases. Because of their physicochemical properties, these substances may be industrially used as cleaning solvents in the electronic components, fire suppression agents in the fire protection, and heat transfer fluids in the heat exchangers. From the environmental, ecological, and healthy points of view, it is urgent to understand their environmental risks of these HFPEs. This article aimed at introducing these HFPEs in physiochemical properties and potential uses, and evaluating their environmental risks (i.e., global warming, photochemical potential, and environmental partition). Further, the updated data on their toxicological profiles and potential exposure hazards from their degradation products were also addressed in this paper. It is indicated that HFPEs still pose some significant hazards, especially global warming and photochemical potentials, to the atmosphere. Regarding the estimation of partition properties (i.e., vapor pressure, octanol-water partition coefficient and bioconcentration) of HFPEs, the predicted values of logKow for several HFPEs were found to be below zero, suggesting that they should possess very low potential for bioaccumulation in the environment. PMID- 17118548 TI - Analysis of adsorption characteristics of 2,4-dichlorophenol from aqueous solutions by activated carbon fiber. AB - In this study experiments were conducted to investigate the adsorption of 2,4 dichlorophenol (2,4-DCP) by activated carbon fiber (ACF) activated by static air. With the results of batch experiments at various temperatures, the adsorption isotherms, kinetics and thermodynamics of this adsorption process were evaluated. Four adsorption isotherm models, Langmuir, Freundlich, Redlich-Peterson and Toth equations, were used to fit the experimental data and the results reveal that the adsorption isotherm models fitted the data in the order of: Langmuir>Redlich Peterson>Toth>Freundlich isotherms. A pseudo second-order adsorption model was better to describe the adsorption data than the pseudo first-order model and the Bangham model at the temperatures tested. The activation energy was calculated to be 40.90 kJ/mol, while the thermodynamic parameters DeltaH and DeltaS were estimated to be -5.82 kJ/mol and 0.07 kJ/(molK), respectively. PMID- 17118549 TI - Fluoride in drinking water and human urine in Southern Haryana, India. AB - The objective of this study was to determine the fluoride content in drinking water and urine samples of adolescent males aged 11-16 years living in Southern Haryana, India. A total of 30 drinking water sources in the studied habitations were assessed for fluoride contamination. Fluoride was estimated in the urine of 400 male children randomly selected from these habitations. The fluoride concentration in drinking water and urine samples was determined using USEPA fluoride ion selective electrode method. The mean fluoride concentration in drinking water samples of Pataudi, Haily Mandi and Harsaru villages was 1.68+/ 0.35, 3.22+/-1.18 and 1.78+/-0.12 mg/l, respectively. The mean urinary fluoride concentration was 2.26+/-0.024 mg/l at Pataudi, 2.48+/-0.77 mg/l at Haily Mandi and 2.43+/-0.84 mg/l at Harsaru village. The higher fluoride levels in the urine of children may be associated to higher fluoride levels in drinking water. The accuracy of measurements was assessed with known addition method in water and urine. Mean fluoride recovery was 98.0 and 99.1% in water and urine. The levels obtained were reproducible with in +/-3% error limit. PMID- 17118551 TI - Societal concerns and risk decisions. AB - Societal concern is a relatively new term and refers to hazards with the capability to generate socio-political responses. Hazards invoking societal concerns pose a challenge to decision makers for they oftentimes have major policy implications yet frequently lack the analytic support affording them such elevated status. Regulators and corporate risk managers, therefore, have been confronted with the difficult question -how and to what extent should societal concerns be factored into risk management decisions? - a question which is compounded by the tendency of 'politicians' to want to accommodate these concerns. Here we first seek to explore the drivers of societal concerns prior to considering the implications for decision makers. We conclude that societal concerns stem from highly disparate causes, are not necessarily originated by the public as is often implied, and as such have markedly differing legitimacies. Furthermore, we note that attempts to incorporate societal concerns into risk decisions raise a host of methodological, political, and ethical issues which suggest, at the very least, that deep caution is required, especially where policy implications are high. PMID- 17118550 TI - Thermodynamics and kinetics of adsorption of Cu(II) onto waste iron oxide. AB - This study investigates low-cost sorbents as replacements for current costly methods of removing heavy metals from solution. This investigation explores the waste iron oxide material (F1), which is a by-product of the fluidized-bed reactor (FBR)-Fenton reaction, for use in the treatment of the wastewater in Taiwan. X-ray powder diffraction (XRD) and scanning electron microscopy (SEM) were used to characterize the F1. In this investigation, F1 are tested as adsorbents for removing copper (Cu(2+)) from aqueous solutions. The highest Cu(2+) adsorption capacity of F1 adsorbent was determined as 0.21 mmolg(-1) for 0.8 mmoldm(-3) initial Cu(2+) concentration at pH 6.0 and 300 K. Adsorption data were well described by the Freundlich model and the thermodynamic constants of the adsorption process, DeltaG degrees , DeltaH degrees and DeltaS degrees were evaluated as -6.12 kJmol(-1) (at 318 K), 9.2 kJmol(-1) and 48.19 Jmol(-1)K(-1) (at 318 K), respectively. Additionally, a pseudo-second-order rate model was adopted to describe the kinetics of adsorption. PMID- 17118552 TI - Kinetics of mercury ions removal from synthetic aqueous solutions using by novel magnetic p(GMA-MMA-EGDMA) beads. AB - Poly(glycidylmethacrylate-methylmethacrylate), p(GMA-MMA-EGDMA), magnetic beads were prepared via suspension polymerization in the presence of ferric ions. The epoxy groups of the beads were converted into amino groups via ring opening reaction of the ammonia and, the aminated magnetic beads were used for the removal of Hg(II) ions from aqueous solution in a batch experiment and in a magnetically stabilized fluidized bed reactor (MFB). The magnetic p(GMA-MMA EGDMA) beads were characterized with scanning electron microscope (SEM), FT-IR and ESR spectrophotometers. The optimum removal of Hg(II) ions was observed at pH 5.5. The maximum adsorption capacity of Hg(II) ions by using the magnetic beads was 124.8+/-2.1 mgg(-1) beads. In the continuous MFB reactor, Hg(II) ions adsorption capacity of the magnetic beads decreased with an increase in the flow rate. The maximum adsorption capacity of the magnetic beads in the MFB reactor was 139.4+/-1.4 mgg(-1). The results indicate that the magnetic beads are promising for use in MFB for removal of Hg(II) ions from aqueous solution and/or waste water treatment. PMID- 17118553 TI - Removal of organic pollutants from 2,2',5,5'-tetrachlorobenzidine (TCB) industrial wastewater by micro-electrochemical oxidation and air-stripping. AB - A feasible method for treatment of the wastewater from the two-staged neutralization in 2,2',5,5'-tetrachlorobenzidine (TCB) manufacturing processes, a refractory dye intermediate effluents, based on combined micro-electrochemical oxidation or iron-chipping filtration (ICF) and air-stripping reactor (ASR), was developed. On conditions of HRT 1h, pH 3.0 in ICF and HRT 38 h, gas-liquid ratio 15, pH 6.0-8.65, temperature 26 degrees C in ASR, the overall COD, color, TCB and NH(4)(+)-N removal were 96.8%, 91%, 87.61% and 62%, respectively, during the treatment of TCB wastewater from the two-staged neutralization dissolved by methanol. The averaged 18.3%, 81.7% of the total degraded COD, 35.2%, 64.8% of TCB were carried out in ICF and ASR, respectively. NH(4)(+)-N removal was finished mainly in ASR. The experimental results indicated that the combined micro-electrochemical oxidation and air-stripping process performed good treatment of COD, color, TCB and NH(4)(+)-N removal in TCB wastewater from the two-staged neutralization dissolved by ethanol or acetone, came up the discharge standard in China. But the TCB wastewater from the two-staged neutralization dissolved by methanol should be deeply treated before discharged. PMID- 17118554 TI - Preparation and XPS studies of macromolecule mixed-valent Cu(I, II) and Fe(II, III) complexes. AB - A new macromolecule ligand and its mixed-valent Cu(I, II) and Fe(II, III) complexes have been prepared by using ethylenediamine as core and maleic anhydride as branched units and characterized by UV-vis, FT-IR, thermal analysis and X-ray photoelectron spectroscopy (XPS). The data obtained from these studies suggested that the coordinate bonds of N-->M, Cl-->M, Ph-OH-->M and H(2)O-->M have been formed and possible binding models are proposed for these complexes. The thermal analysis (TG-DTG) reveals that these complexes possess thermal stable property below 800 degrees C. PMID- 17118555 TI - Electrodialytic extraction of Cd and Cu from sediment from Sisimiut Harbour, Greenland. AB - A previous study showed that the sediment of Sisimiut Harbour, Greenland is polluted with Cu and Cd to an extent where toxicological effects must be expected. This study was aimed at evaluating the possibility for removing Cu and Cd from this sediment by an electrodialytic method and also to evaluate the removal rate of the two heavy metals. The sediment was suspended in distilled water during application of current. Both heavy metals were removed successfully. The Cu concentration was reduced from 97 to 16mg/kg and the Cd concentration was reduced from 0.55 to 0.03mg/kg after 28 days with an applied current density of 1.2mAcm(-2). However, it was seen that the removal rate decreased considerably after 3 days with an applied current density of 0.5mAcm(-2) and the major part of the two heavy metals was removed during the first 3 days (the Cu concentration was reduced with 74% and the Cd concentration with 80%). Thus a large reduction in concentrations can be obtained relatively fast. During the process of electrodialytic treatment the sediment suspension is acidified and reaches pH 2 after about 3 days (with 0.5mAcm(-2)), where it stabilizes. A comparison with chemical extraction in HNO(3) at about the same pH showed that 50-70% more Cu and 11% more Cd was removed during the electrodialytic treatment. PMID- 17118556 TI - The amygdala: different pains, different mechanisms. PMID- 17118557 TI - A serological study of Babesia caballi and Theileria equi in Thoroughbreds in Trinidad. AB - Ninety-three (93) horses were investigated for serum antibodies to Theileria equi (T. equi) and Babesia caballi (B. caballi) using the immunofluorescent antibody test (IFAT). Seventy-seven (82.8%) horses were seropositive; 31 (33.3%) were positive to T. equi compared to 64 (68.8%) to B. caballi while 18 (19.4%) horses were seropositive to both parasites. No significant differences in antibody frequencies among females and males for either T. equi or B. caballi were noted. Differences in seropositivity to B. caballi among age groups were not significant. Antibodies to T. equi were more frequent than to B.caballi in the age group 5 years and over than in the 1-2 and 2-4 years age groups (p<0.05). Unlike T. equi antibodies, B. caballi antibodies in horses in the county of Caroni were significantly less frequent when compared to other counties (p<0.05). Of 18 (19.4%) clinically ill horses, seven (42.9%) had clinicopathological evidence of anemia. Only one-third (6 of 18) horses were positive for the parasite on Wright-Giemsa stained blood smears and anemia was present in only 2. We report here that B. caballi and not T. equi may be the more common agent of piroplasmosis in Trinidad. PMID- 17118558 TI - In vivo effects of chronic contamination with depleted uranium on vitamin D3 metabolism in rat. AB - The extensive use of depleted uranium (DU) in today's society results in the increase of the number of human population exposed to this radionuclide. The aim of this work was to investigate in vivo the effects of a chronic exposure to DU on vitamin D(3) metabolism, a hormone essential in mineral and bone homeostasis. The experiments were carried out in rats after a chronic contamination for 9 months by DU through drinking water at 40 mg/L (1 mg/rat/day). This dose corresponds to the double of highest concentration found naturally in Finland. In DU-exposed rats, the active vitamin D (1,25(OH)(2)D(3)) plasma level was significantly decreased. In kidney, a decreased gene expression was observed for cyp24a1, as well as for vdr and rxralpha, the principal regulators of CYP24A1. Similarly, mRNA levels of vitamin D target genes ecac1, cabp-d28k and ncx-1, involved in renal calcium transport were decreased in kidney. In the brain lower levels of messengers were observed for cyp27a1 as well as for lxrbeta, involved in its regulation. In conclusion, this study showed for the first time that DU affects both the vitamin D active form (1,25(OH)(2)D(3)) level and the vitamin D receptor expression, and consequently could modulate the expression of cyp24a1 and vitamin D target genes involved in calcium homeostasis. PMID- 17118559 TI - Immobilized alpha2,6-linked sialic acid suppresses caspase-3 activation during anti-IgM antibody-induced apoptosis in Ramos cells. AB - In Ramos cells, a human Burkitt's lymphoma cell line, stimulation of the B cell antigen receptor with anti-IgM antibody (Ab) induces apoptosis as indicated by a decrease in cell viability and an increase in DNA fragmentation and cell surface exposure of phosphatidylserine. Furthermore, these changes are suppressed by incubating the cells in alpha(1)-acid glycoprotein (AGP)-coated tissue culture plates. Here, we found that, during Anti-IgM Ab-induced apoptosis in Ramos cells, caspase-3 is activated downstream of caspase-8 and the mitochondrial pathway is activated, as indicated by a loss of mitochondrial membrane potential, an increase in the release of cytochrome c to the cytoplasm, and enhanced Bax expression. Anti-IgM Ab-induced apoptosis of neuraminidase-treated Ramos cells was suppressed by incubating the cells on plates coated with AGP, which contains a high concentration of alpha2,6-linked sialic acid. The incubation on plates coated with AGP also suppressed anti-IgM Ab-stimulated caspase-3 activity and increased the level of X-linked inhibitor of apoptosis protein (XIAP), but it did not affect caspase-8 activity, the mitochondrial membrane potential, cytochrome c release, or Bax expression. The results indicate that the interaction of Ramos cells with immobilized alpha2,6-linked sialic acid enhances XIAP expression, directly or indirectly suppressing caspase-3 activity and inhibiting anti-IgM Ab induced apoptosis. PMID- 17118560 TI - Effect of thiocyanate on the peroxidase and pseudocatalase activities of Leishmania major ascorbate peroxidase. AB - We report here that the Leishmania major ascorbate peroxidase (LmAPX), having similarity with plant ascorbate peroxidase, catalyzes the oxidation of suboptimal concentration of ascorbate to monodehydroascorbate (MDA) at physiological pH in the presence of added H(2)O(2) with concurrent evolution of O(2). This pseudocatalatic degradation of H(2)O(2) to O(2) is solely dependent on ascorbate and is blocked by a spin trap, alpha-phenyl-n-tert-butyl nitrone (PBN), indicating the involvement of free radical species in the reaction process. LmAPX thus appears to catalyze ascorbate oxidation by its peroxidase activity, first generating MDA and H(2)O with subsequent regeneration of ascorbate by the reduction of MDA with H(2)O(2) evolving O(2) through the intermediate formation of O(2)(-). Interestingly, both peroxidase and ascorbate-dependent pseudocatalatic activity of LmAPX are reversibly inhibited by SCN(-) in a concentration dependent manner. Spectral studies indicate that ascorbate cannot reduce LmAPX compound II to the native enzyme in presence of SCN(-). Further kinetic studies indicate that SCN(-) itself is not oxidized by LmAPX but inhibits both ascorbate and guaiacol oxidation, which suggests that SCN(-) blocks initial peroxidase activity with ascorbate rather than subsequent nonenzymatic pseudocatalatic degradation of H(2)O(2) to O(2). Binding studies by optical difference spectroscopy indicate that SCN(-) binds LmAPX (Kd = 100 +/- 10 mM) near the heme edge. Thus, unlike mammalian peroxidases, SCN(-) acts as an inhibitor for Leishmania peroxidase to block ascorbate oxidation and subsequent pseudocatalase activity. PMID- 17118561 TI - A simple and effective procedure for treating burn contractures: releasing incision and quadra Z technique. AB - Burn contractures particularly involving the joints are challenging problems which might cause severe functional impairments. Many surgical techniques have been described for use, however, an ideal method yet to be found. Releasing incision is the most common and effective way to release the wide and severe contractures but it has some drawbacks. We propose a releasing incision technique combined with four Z plasty incisions to overcome the disadvantages of traditional releasing incision technique. We successfully used our releasing incision and quadra Z technique on seven consecutive patients with burn contractures between 2003 and 2005. We modified the classical releasing incision technique by adding four Z plasties; two of them with a common base on each corner of the incision line. In this technique, limitation of the webbing following the incision is made possible by the transposed flaps and unnecessary lateral extension of the incision and the defect was avoided, i.e. maximum release gain with minimal defect was provided. Satisfactory results were achieved in seven patients treated with this technique due to significant burn contractures between 2003 and 2005 with no significant complication. We propose this technique is suitable in all patients with severe burn contractures who require releasing incision and grafting. PMID- 17118562 TI - Antithrombin deficiency and its relationship to severe burns. AB - Antithrombin (AT) is an important endogenous anticoagulant and exhibits marked anti-inflammatory properties. To evaluate the incidence of AT deficiency in severe burn and its correlation to the variables of the abbreviated burn severity index (ABSI), length of hospital stay (LOS) and mortality we collected data on the substitution of human plasma-derived AT concentrate in 201 consecutive patients suffering from severe burn. One hundred and eight patients (54%) developed AT deficiency during their hospitalisation and, according to our institutional practice, received substitution therapy by continuous infusion to maintain physiological plasma activity (70-120%). The mean administered dose served as a measure of AT deficiency. The percentage of patients in an AT deficient state was highest within the first 5 days after injury. It was 26% on day 1 and between 38% and 41% on days 2-5 and thereafter decreased constantly over time. A multiple regression analysis between the dependent variable mean administered dose of AT concentrate and the independent variables age, total body surface area burned (TBSA), gender, inhalation injury (INHAL), full thickness burn (FTB), LOS and mortality was performed. Age, gender and FTB showed no significant influence on the development of AT deficiency. Increasing TBSA and INHAL clearly increase the risk of developing AT deficiency (p-values 0.0001 and 0.037). The analysis also identified AT deficiency as an independent predictor of LOS and mortality (p-values 0.036 and 0.003). Development of AT deficiency is a frequent event after burn with significant correlation to TBSA and INHAL, increased mortality rates and longer hospital stays. PMID- 17118563 TI - Properties of matter matter in assessment of scald injuries. AB - Gathering information on the thermal characteristics of the causative agent in scald injuries provides clues as to the likely depth of injury. We theorize that viscosity and thermal capacity may have important roles to play when we view scalds as contact burns due to a liquid. From a 4-year review of our scald patients, we found that scalds due to thick food/drinks such as congee (a porridge made from rice) were associated with a higher rate of surgery. We determined the rate of cooling of seven common food/drinks and found little difference between water, tea, coffee and noodles, other than the starting temperatures. However, the rate of cooling of congee was significantly slower indicating a greater thermal capacity. A "drip" model found that a skin substitute exposed to congee cooled significantly more slowly compared to other food/drinks, suggesting that its greater viscosity plays a role. This supports the theory that the viscosity of food/drink is important. PMID- 17118564 TI - A clinical case study of a participation intervention method for burn dressing change in two children. PMID- 17118565 TI - Hemispheric differences in protein kinase C betaII levels in the rat amygdala: baseline asymmetry and lateralized changes associated with cue and context in a classical fear conditioning paradigm. AB - The amygdala is critically important for fear learning, and specific kinases have been implicated as contributors to the mechanisms that underlie learning. We examined levels of protein kinase C betaII (PKC betaII) in the left and right lateral and basolateral nuclei (LA/BLA) of the amygdala from animals that were classically fear conditioned with tones as cues and footshocks. Groups consisted of animals that received neither tones nor shocks, paired tones and shocks, or unpaired tones and shocks. At 1 h after conditioning, some animals from each group were used for biochemical measurements of PKC betaII levels and other animals were given probe trials to assess freezing behavior to cue and context. The levels of PKC betaII were greater in the left hemisphere in animals receiving neither tones nor shocks and animals receiving paired tones and shocks. PKC betaII levels were greater in the right hemisphere of animals receiving randomly presented tones and shocks. Freezing times to cue were long (>80% of probe trial time) in both the paired tone/shock and randomly unpaired tone/shock groups. Freezing times to context were long in the unpaired tone/shock group, but not the paired tone/shock group. Correlational analyses showed that freezing times to context, but not cue, precisely predicted the right/left relation of PKC betaII levels in the LA/BLA: the greater the time spent freezing to context, the greater the increase in right hemisphere PKC betaII levels. We conclude that fear conditioning causes hemisphere and input specific increases in PKC betaII in the rat LA/BLA. PMID- 17118566 TI - Repeated long separations from pups produce depression-like behavior in rat mothers. AB - Long maternal (LMS) versus brief maternal (BMS) daily separations of rat pups from their mothers have contrasting effects on their adult stress responses and maternal behavior by, respectively, decreasing and increasing licking received from their mothers. We hypothesized that LMS decreases pup-licking in mothers by inducing learned helplessness, creating a depression-like state. We subjected postpartum rats to LMS (3 h), BMS (15 min) or no separation (NMS) on postpartum days 2-14. After weaning, mothers were given a forced swim test (FST). LMS mothers exhibited more immobility and fewer escape attempts than BMS or NMS mothers. These results suggest that LMS induces a depression-like state, which may account for the reductions in maternal behavior seen in LMS mothers. Immobility in the FST is recognized as an animal model of depression. Therefore, LMS may be a model of maternal depression. PMID- 17118567 TI - Towards a better understanding of the aetiology of Legg-Calve-Perthes' disease: acetabular retroversion may cause abnormal loading of dorsal femoral head-neck junction with restricted blood supply to the femoral epiphysis. AB - The most accepted part of the aetiology of Perthes' disease is an intermittent arterial occlusion. Many different reports now also suggest that some mechanical factor plays a role. In search of mechanical differences between normal hips and Perthes hips, many secondary morphologic differences of the proximal femur and the acetabulum are known. Recent research by the author has shown morphological changes of the acetabulum, independent of Perthes' disease in a large part of patients. In particular, retroversion of the acetabulum was found in a large part of adults who suffered from Perthes' disease as a child, and even in children who just developed Perthes' disease. The author suggest a relation of the two entities and hypotheses that due to the acetabular retroversion, there may be an intermittent, movement depending abnormal loading pressure on the dorsal femoral head-neck junction and the local blood vessels, which can lead to insufficient blood supply of the femoral epiphysis and to local necrosis, as these vessels are the only blood supply to the femoral epihysis in children. PMID- 17118568 TI - How does thrombolysis increase defibrillation success? PMID- 17118569 TI - The role of palliative radiation therapy in symptomatic locally advanced gastric cancer. AB - PURPOSE: To review the outcome of palliative radiotherapy (RT) alone in patients with symptomatic locally advanced or recurrent gastric cancer. METHODS AND MATERIALS: Patients with symptomatic locally advanced or recurrent gastric cancer who were managed palliatively with RT at The Cancer Institute, Singapore were retrospectively reviewed. Study end points included symptom response, median survival, and treatment toxicity (retrospectively scored using the Common Toxicity Criteria v3.0 [CTC]). RESULTS: Between November 1999 and December 2004, 33 patients with locally advanced or recurrent gastric cancer were managed with palliative intent using RT alone. Median age was 76 years (range, 38-90 years). Twenty-one (64%) patients had known distant metastatic disease at time of treatment. Key index symptoms were bleeding (24 patients), obstruction (8 patients), and pain (8 patients). The majority of patients received 30 Gy/10 fractions (17 patients). Dose fractionation regimen ranged from an 8-Gy single fraction to 40 Gy in 16 fractions. Median survival was 145 days, actuarial 12 month survival 8%. A total of 54.3% of patients (13/24) with bleeding responded (median duration of response of 140 days), 25% of patients (2/8) with obstruction responded (median duration of response of 102 days), and 25% of patients (2/8) with pain responded (median duration of response of 105 days). No obvious dose response was evident. One Grade 3 CTC equivalent toxicity was recorded. CONCLUSION: External beam RT alone is an effective and well tolerated modality in the local palliation of gastric cancer, with palliation lasting the majority of patients' lives. PMID- 17118570 TI - Association between pathologic response in metastatic lymph nodes after preoperative chemoradiotherapy and risk of distant metastases in rectal cancer: An analysis of outcomes in a randomized trial. AB - PURPOSE: To compare 5 x 5 Gy preoperative radiotherapy with immediate surgery vs. preoperative chemoradiotherapy (50.4 Gy, 5-fluorouracil, leucovorin) with delayed surgery in a randomized trial for cT3-T4 low-lying rectal cancer. Despite the downstaging effect of chemoradiotherapy, similar long-term outcomes were observed in both groups. METHODS: The Cox model was used to evaluate the prognostic value of ypTN ("yp" denotes that pathologic classification was performed after initial multimodality therapy) categories and the surgical margin status in 291 patients. RESULTS: Disease-free survival (DFS) (hazard ratio [HR] 1.05, 95% confidence interval [CI], 0.73-1.51), distant metastases (HR, 1.17; 95% CI, 0.77-1.78), and local control (HR, 1.45; 95% CI, 0.74-2.84) were similar in both arms. The ypN status was the only independent prognostic factor for DFS (p < 0.001). An interaction (p = 0.016) between N stage and the assigned treatment was demonstrated. For ypN-negative patients, DFS was similar in both arms (HR, 0.83, 95% CI, 0.47-1.48); however, for ypN-positive patients, DFS was worse in the chemoradiotherapy arm (HR, 1.73; 95% CI, 1.07-2.77). The 4-year (median follow up) DFS rate in N-positive patients was 51% in the 5 x 5-Gy arm vs. 25% in the chemoradiotherapy arm. The corresponding 4-year rates for the incidence of local recurrence and distant metastases were 14% vs. 27% (HR, 1.95; 95% CI, 0.78-4.86) and 38% vs. 68% (HR, 2.05; 95% CI, 1.21-3.48). CONCLUSION: N-positive disease after chemoradiotherapy indicates radiochemoresistance. N-positive disease after 5 x 5 Gy RT includes both radiosensitive and radioresistant tumors, because the interval between radiotherapy and surgery was too short for radiosensitive cancer to undergo necrosis. Thus, the greater risk of distant metastases recorded in the chemoradiotherapy arm suggests that radiochemoresistance of nodal metastases from rectal cancer is associated with a high potential for developing distant metastases. PMID- 17118571 TI - Neoadjuvant chemotherapy for radioinduced osteosarcoma of the extremity: The Rizzoli experience in 20 cases. AB - PURPOSE: Evaluate treatment and outcome of 20 patients with radioinduced osteosarcoma (RIO). Because of previous primary tumor treatment, RIO protocols were different from others we used for non-RIO. PATIENTS AND METHODS: Between 1983 and 1998, we treated 20 RIO patients, ages 4-36 years (mean 16 years), with chemotherapy (two cycles before surgery, three postoperatively). The first preoperative cycle consisted of high-dose Methotrexate (HDMTX)/Cisplatinum (CDP)/Adriamycin (ADM) and the second of HDMTX/CDP/Ifosfamide (IFO). The three postoperative treatments were performed with cycles of MTX/CDP; IFO was used as single agent per cycle repeated three times. RESULTS: Two patients received palliative treatment because their osteosarcoma remained unresectable after preoperative chemotherapy. The remaining 18 patients had surgery (7 amputations, 11 resections); histologic response to preoperative chemotherapy was good in 8 patients, poor in 10. At a mean follow-up of 11 years (range, 7-22 years), 9 patients remained continuously disease-free, 10 died from osteosarcoma and 1 died from a third neoplasm (myeloid acute leukemia). These results are not significantly different from those achieved in 754 patients with conventional osteosarcoma treated in the same period with protocols used for conventional treatment. However, this later group had an 18% 3-year event-free survival after treatment of relapse vs. 0% in the RIO group. CONCLUSION: Treated with neoadjuvant chemotherapy RIO seem to have an outcome that is not significantly different from that of comparable patients with conventional primary high grade osteosarcoma (5-year event-free survival: 40% vs. 60%, p = NS; 5-year overall survival 40% vs. 67%, p < 0.01). PMID- 17118572 TI - Antiinflammatory activity of the aqueous leaf extract of Byrsocarpus coccineus. AB - The antiinflammatory effect of the aqueous leaf extract of Byrsocarpus coccineus was evaluated using the carrageenan and egg albumin induced rat paw edema, xylene induced mouse ear edema and formaldehyde induced arthritis inflammation tests. The extract administered orally at doses of 50, 100, 200 and 400 mg/kg b.w produced a significant (P<0.05) dose dependent inhibition of edema formation in all four methods used. The results obtained suggest that the aqueous leaf extract of B. coccineus is endowed with effective antiinflammatory activity mediated via either inhibition of phospholipase A(2) (PLA(2)) activity or cyclooxygenase cascade and by blocking the release of vasoactive substances (histamine, serotonin and kinins). These findings seem to justify the use of the plant in traditional African medicine in the treatment of inflammation, including arthritic conditions. PMID- 17118573 TI - Testing for episodic-like memory in rats in the absence of time of day cues: replication of Babb and Crystal. AB - Two experiments were performed to look for evidence of episodic-like memory in rats. On each of a series of trials on an eight-arm radial maze, rats in two groups entered four open arms in Phase 1, with reward pellets on three arms and a favored reward (chocolate in Experiment 1 and cheese in Experiment 2) on the remaining arm. Phase 2 retention tests were given 30 min or 4 h after Phase 1, with all eight arms open. The four arms not entered in Phase 1 all contained reward pellets, and the three arms that contained pellets in Phase 1 were empty. In the replenish short group, the favored reward was replenished at the same location where it was found in Phase 1 at the 30 min retention interval but was absent (Experiment 1) or degraded (Experiment 2) at the 4 h retention interval. In the replenish long group, the favored reward was replenished at the 4 h retention interval but not at the 30 min retention interval. Over a number of daily trials that randomly mixed short and long delays, rats in both experiments learned to return earlier to the arm containing the favored reward at the retention interval when it was replenished than at the retention interval when it was absent or degraded. These results replicate earlier findings [Babb, S.J., Crystal, J.D., 2005, Discrimination of what, when, and where: implications for episodic-like memory in rats. Learn. Mot., 36, 177-189] and provide evidence of episodic-like memory in rats. PMID- 17118574 TI - Social and sexual behaviours aid transmission of bacteria in birds. AB - Understanding the behavioural mechanisms that mediate pathogen transmission in social hosts like birds could provide the empirical bases for explaining the epidemiological dynamics of zoonotic infections in vertebrates. By experimentally infecting the feathers and cloaca of captive zebra finches (Taeniopygia guttata), with the bacterium Bacillus licheniformis PWD1 (BL), we examined the self contamination and horizontal transmission of birds sharing the same environment. We also examined whether sexual transmission of bacteria is gender biased. Our results show that bacteria placed on the plumage of the birds lead to self and allo-infections of the bird guts, possibly through preening behaviours and bacterial ingestion. Furthermore, we found that sexual transmission of the bacteria was asymmetrical, being higher when males are the transmitting sex. Our results suggest the existence of an oral-faecal-genital route of bacterial transmission for avian hosts, wherein bacteria present on feathers infect their host guts through self and allo-preening and bacterial ingestion. Gut bacteria can then be transmitted sexually with transmission rate being higher when males are the infected sex. PMID- 17118575 TI - Endogenous oscillations in short-interval timing. AB - A defining feature of a circadian oscillator is that periodic output from the oscillator continues after the termination of periodic input. In contrast, a defining feature of a pacemaker-accumulator system is that elapsed time is measured with respect to the presentation of a stimulus, according to the classic description of this system. Consequently, the output of a short-interval system is periodic if presented with periodic input, but periodic output can be expected to cease if periodic input is discontinued. Groups of rats were trained to time short intervals (1-3 min); periodic delivery of food produced periodic behavior. Next, delivery of food was suspended. Behavior was periodic after termination of periodic input, and the period in extinction increased as a function of the period in training. These data suggest that short-interval timing is, at least in part, based on a self-sustaining, endogenous oscillator. PMID- 17118576 TI - Development and validation of the Stimulant Relapse Risk Scale for drug abusers in Japan. AB - OBJECTIVE: To develop and validate a multidimensional measure of relapse risk for stimulants in Japanese drug abusers. METHODS: A Stimulant Relapse Risk Scale (SRRS) was developed based on the Marijuana Craving Questionnaire and a discussion among three psychiatrists. We created 48 items after confirming the items including a variety of relapse risk, such as craving (expectancy, compulsivity, etc.) and emotionality problems. One hundred inpatients and outpatients with a history of stimulant abuse (71 males and 29 females) were recruited with informed consent, and were administered the SRRS. The Visual Analogue Scale for drug craving (VAS), Addiction Severity Index for Japanese (ASI J), and data on relapse within 3 and 6 months after the rating were used for the validation. RESULTS: Exploratory factor analysis highlighted five factors: anxiety and intention to use drug (AI), emotionality problems (EP), compulsivity for drug use (CD), positive expectancies and lack of control over drug (PL), and lack of negative expectancy for drug use (NE). These accounted for 48.3% of the total variance. Thirty of the 43 items were classified into the five subscales. Cronbach's alpha coefficient for each subscale ranged from .55 to .82, and was .86 for the total SRRS, indicating their adequate internal consistency. AI, CD, PL, and total SRRS were significantly correlated with the drug-use composite score of the ASI-J, supporting their concurrent validity. AI, PL, NE, and total SRRS were significantly correlated with relapse, implying their predictive validity. CONCLUSIONS: The SRRS has multidimensional psychometric properties useful for assessing the various aspects of stimulant relapse risk. PMID- 17118577 TI - Effects of smoking cessation on health care use: is elevated risk of hospitalization among former smokers attributable to smoking-related morbidity? AB - BACKGROUND: Although the association between smoking status and health services use is well established, this relation is not well-studied for the comparison of current and former smokers. Some studies showed higher utilization of health services among former smokers compared to continuing smokers. This study investigates the relation between smoking status, time since smoking cessation and hospitalization in a general population sample. We hypothesized that elevated risk of hospitalization among former smokers compared with continuing smokers is related to higher smoking-related morbidity among former smokers. METHODS: Data from a cross-sectional sample of 4310 adults aged 20-79 in Pomerania, Germany was used (response proportion 68.8%). Smoking status, time since smoking cessation (in years), and date of diagnosis of smoking-related diseases were determined from self-reports. We used fractional polynomials to determine the dose-response relation of time since cessation and risk of hospitalization. Confounding was investigated allowing for different sets of confounding variables. RESULTS: We found that the probability of hospitalization was highest among those who quit 1 3 years ago and decreased thereafter. Adjustment for health status and socio economic variables revealed that this association is attenuated by current diagnosis of smoking-related diseases. CONCLUSION: Short-term excess health care utilization among former smokers might result from smoking-related conditions that may have led to smoking cessation. Findings suggest that smoking cessation has long-term health benefits resulting in lower health care needs. PMID- 17118578 TI - Stigma, discrimination and the health of illicit drug users. AB - Persons who use illicit drugs are stigmatized in the United States. The conferral of a deviant social status on illicit drug users may serve to discourage use. However, stigmatization may also adversely affect the health of those who use illicit drugs, through exposure to chronic stress such as discrimination and as a barrier to accessing care. We hypothesized that aspects of stigma and discrimination would be associated with mental and physical health among illicit drug users. Using street outreach techniques, 1008 illicit drug users were interviewed about stigma and discrimination related to their drug use, and their health. We measured discrimination related to drug use, alienation, perceived devaluation, and responses to discrimination and stigma. Health measures included mental and physical health measures from the Medical Outcomes Study Short Form 36, depression symptoms from the Center for Epidemiological Studies Depression scale, and a sum of health conditions. In adjusted models, discrimination and alienation were both associated with poorer mental health, and only discrimination was associated with poorer physical health. Angry responses to discrimination and stigma were associated with poorer mental health. The association of stigma and discrimination with poor health among drug users suggests the need for debate on the relative risks and benefits of stigma and discrimination in this context. PMID- 17118579 TI - Profiling of maternal and developmental-stage specific mRNA transcripts in Atlantic halibut Hippoglossus hippoglossus. AB - cDNA libraries were constructed from the following developmental stages (tissues) of the Atlantic halibut (Hippoglossus hippoglossus): 2-cell stage (embryos), 1 day-old yolk sac larvae (trunk) and juvenile (fast skeletal muscle). A total of 4249 high quality expressed sequence tags from the three libraries were clustered into a partial transcriptome of 2124 putative genes. A large proportion of the gene clusters (48.3%) had no significant matches against known proteins. The most abundant ESTs of nuclear transcripts in the 2-cell library included sequences with high identity to zebrafish H1M, a linker histone-like protein involved in primordial germ cell specification, zinc finger protein, rRNA external transcribed spacer, thymosin beta-4, cyclin B1 and several predicted peptides from the Tetraodon nigroviridis genome assembly with unknown functions. 170 and 123 ESTs represented ribosomal proteins in the larval and juvenile libraries respectively, compared with only two sequences in the 2-cell library, which may reflect an abundance of maternally inherited pre-formed ribosomes in the yolk. Even though some clusters were common to all three libraries, most putative genes showed a developmental-stage specific distribution with 72% (2-cell embryo), 59% (larval) and 57% (juvenile) sequences having no significant matches against the 8400 adult halibut sequences in the EMBL nucleotide database. Comparison between the predicted halibut peptide data set and the human, zebrafish, and pufferfishes (T. nigroviridis and Takifugu rubripes) proteomes revealed that, as expected, the halibut sequences were more similar to the other two fish species than to human proteins. However, no clear bias towards the pufferfishes was observed, suggesting significant sequence variation between orthologues within the clade Acanthomorpha. The sequence information generated in the present study will represent a significant new resource for future studies on normal and abnormal development in Atlantic halibut. PMID- 17118580 TI - Functional characterization of the promoter region of the chicken elongation factor-2 gene. AB - Elongation factor 2 (EF-2) plays a key role in the essential process of protein synthesis by translocating tRNAs from the ribosomal A- and P-sites to the P- and E-sites. EF-2 regulates the outcome of protein synthesis in mammalian cells. This report demonstrates that chicken EF-2 protein levels are dependent on transcription in 8-bromo-cAMP, insulin and phorbol ester-treated cells. In order to delineate functional domains that control chicken EF-2 gene transcription, the 5'-flanking region of the chicken EF-2 promoter was analyzed. Deletion constructs from -550 and -86 had the same basal level promoter activity as the whole EF-2 promoter. The sequence between nucleotides -700 and -550 was determined to be a regulatory region for the chicken EF-2 basal promoter activity. The region between -700 and -550 has a negative regulatory region and two regulatory proteins (I, II). 8-bromo-cAMP increased chicken EF-2 promoter activity ( 700/+102) in Rat 1 HIR fibroblast cells more than insulin and phorbol ester treatment. Binding of protein I and II were decreased by 8-bromo-cAMP but restored by a protein kinase A inhibitor (KT5720). GATA consensus sequence oligonucleotide and fragment -86/-50 prevented protein II binding of fragment 700/-550. This result suggested that protein II is a GATA-like protein. These observations provide a novel regulatory mechanism for the EF-2 promoter. PMID- 17118581 TI - Mitochondrial genomes from major lizard families suggest their phylogenetic relationships and ancient radiations. AB - In placental mammals and birds, molecular data generally support a view that they diverged into their ordinal groups in good response to mid-Cretaceous continental fragmentations. However, such divergence patterns have rarely been studied for reptiles for which phylogenetic relationships among their major groups have not yet been established molecularly. Here, I determined complete or nearly complete mitochondrial DNA sequences from seven lizard families and reconstructed phylogenetic relationships between major lizard families. When snakes were included, maximum likelihood analysis did not support a morphological view of the snakes-varanoids affinity, although several other competing hypotheses on the position of snakes still cannot be discriminated presumably due to extremely long branches of the snake lineages. I also conducted clock-free Bayesian analyses to show that divergence times between major lizard families were centered in Triassic-Jurassic times. Thus, lizards include much deeper divergences than the mammals and birds and they appear to have already radiated into various families prior to the mid-Cretaceous major continental fragmentation. PMID- 17118582 TI - Most recent AluY insertions in human gene introns reduce the content of the primary transcripts in a cell type specific manner. AB - Being the most effectively transposed primate-specific SINEs, Alu elements are present in more than one million copies in the human genome and include most recently transposed subsets of AluY elements that are polymorphic in humans. Although Alu elements are commonly thought to play an essential role in shaping and functioning of primate genomes, the understanding of the impact of recent Alu insertions on human gene expression is far from being comprehensive. Here we compared hnRNA contents for allele pairs of genes heterozygous for AluY insertions in their introns in human cell lines of various origins. We demonstrated that some AluY insertions correlated with decreased content of the corresponding hnRNAs. The effect observed does not depend on sequences of Alu elements and their orientation but is likely to be cell type specific. PMID- 17118583 TI - Evaluation of three serological tests for diagnosis of Maedi-Visna virus infection using latent class analysis. AB - Maedi-Visna virus (MVV) infection in sheep is present in several European countries, including Norway. The current Norwegian surveillance and control programme for MVV infection uses three serological tests: an agar gel immunodiffusion test (AGID) and two commercially available indirect ELISAs (Institut Pourquier, P-ELISA and HYPHEN BioMed, H-ELISA). From 18 flocks with suspected or confirmed MVV infection, sera from naturally infected sheep were obtained, and sensitivity (Se) and specificity (Sp) of the three tests were estimated in absence of a perfect reference test using latent class models in a Bayesian analysis. The AGID had higher Sp (95% posterior credibility interval (PCI) [98.4; 99.9]) than either ELISA (95% PCI: P-ELISA, [95.1; 99.0]; H-ELISA, [91.4; 96.6]), but much lower Se (95% PCI: AGID, [41.4; 59.8]; P-ELISA, [92.7; 100.0]; H-ELISA, [90.9; 99.4]). Currently the P-ELISA is used for screening and positive samples are subsequently confirmed by a setup using all three tests in a serial reading. The Se and Sp of the serial interpretations with and without the H-ELISA were estimated. The results suggested that the H-ELISA could be dropped as a confirmatory test as the Se of the three test serial reading was reduced significantly without adding a significant improvement of the Sp compared to the serial reading of the P-ELISA and AGID alone. However, the perceived cost of false positives versus false negatives will influence this decision. Estimates of the predictive values for the tests and combinations suggested that the P-ELISA is a good choice of screening, but confirmatory tests are needed to achieve acceptable levels of positive predictive values. PMID- 17118584 TI - Evidence of existence of infectious hypodermal and hematopoietic necrosis virus in penaeid shrimp cultured in China. AB - Infectious hypodermal and hematopoietic necrosis virus is the causative agent of a shrimp disease which causes economic losses on a global scale. A pair of primers, I2814F/I3516R, was designed from the IHHNV genomic sequence (GenBank) that encodes for structural protein corresponding to nucleotides 2814-3516, which amplifies a 703 base pair (bp) region from the virus genome. PCR amplification with the primers generated a product of the expected size from the purified IHHNV DNA of Litopenaeus vannamei and IHHNV-infected penaeid populations but not from the IHHNV-free shrimp, white spot syndrome virus (WSSV) and hepatopancreatic parvovirus (HPV). The PCR amplicon described above was labeled with digoxigenin (DIG)-11-dUTP as a probe used for dot blot hybridization and in situ hybridization test. Under the optimized PCR conditions, the primers were detected by as little as 20 fg of purified IHHNV DNA, which contained only 8.83 x 10(3) copies of IHHNV, a 1000-fold greater than using dot blot hybridization. Sections of histopathology showed eosinophilic intranuclear inclusions (Cowdry type A inclusions or CAIs) in infected tissues while in situ hybridization, cells displayed an intense reaction with the DIG-labeled probe. PCR assay was developed to detect IHHNV in penaeid shrimp and other crustaceans from the rearing ponds of China (March 2001-June 2004). The positive rate was 51.5% (154 out of 299) and 8.3% (2 out of 24) for penaeid shrimp and crab samples, respectively. The survey demonstrated the presence of IHHNV in China. PMID- 17118585 TI - Fluorescence polarization assay for the detection of antibodies to Mycobacterium bovis in bovine sera. AB - The performance of a fluorescence polarization assay (FPA) that detects antibodies to Mycobacterium bovis in bovine sera is described. The FPA reported here is a direct binding primary screening assay using a small polypeptide derived from the M. bovis MPB70 protein. A secondary inhibition assay confirms suspect or presumed positive samples. Specificity studies involved five different veterinary laboratories testing 4461 presumed negative bovine samples. FPA specificity was 99.9%. The FPA was used to identify herd status as either M. bovis infected or non-infected. Herd surveillance studies (nine herds) were performed in Mexico and South Africa. The FPA had a specificity of 100% (two negative herds), and correctly identified six of seven infected herds. Finally, sera from 105 slaughter animals that had gross lesions in lymph nodes similar to those seen with bovine tuberculosis were tested by the FPA. Thin sections from the associated formalin-fixed paraffin-embedded samples of lymph nodes were stained using hematoxylin and eosin (H&E) for morphologic examination and using the Ziehl-Neelsen (ZN) method for detection of acid-fast bacilli. Of the 105 animals, 78 were classified as TB suspect based on lesion morphology, 21 were positive by ZN, 9 were positive by FPA and 13 were positive by PCR for the tuberculosis group of Mycobacterium. Among the 21 ZN positives, 11 (52.4%) were PCR positive. Among the 9 FPA positives, 8 (88.9%) were PCR positive. For the 13 PCR positives, 8 (61.5%) were FPA positive and 11 (84.6%) were ZN positives. These results show that use of the FPA for detection of M. bovis infection of cattle has value for bovine disease surveillance programs. PMID- 17118586 TI - Synchronization and superovulation of mature cycling gilts for the collection of pronuclear stage embryos. AB - An efficient protocol was developed to synchronize and superovulate mature pigs for the collection of pronuclear stage embryos suitable for DNA microinjection. A timed and coordinated regimen of Lutalyse, PG600 and Chorulon along with daily checking for estrus allowed synchronization of groups of gilts having estrous cycles at regular intervals. Pigs 10-16 days after the beginning of standing estrus have been successfully synchronized into estrus using this protocol. A standard dose of each drug was used independent of size or age of the animal. One protocol averaged 38.9 ovulations and 31.1 one-cell embryos recovered per animal. PMID- 17118587 TI - Solution studies on binary and ternary complexes of copper(II) with some fluoroquinolones and 1,10-phenanthroline: Antimicrobial activity of ternary metalloantibiotics. AB - Interaction of norfloxacin and ofloxacin with copper(II) and copper(II)/phenanthroline has been studied in aqueous solution and the stability constants of the binary complexes Cu(II)/fluoroquinolone and of the ternary complexes Cu(II)/phenanthroline/fluoroquinolone have been determined by potentiometry and UV-vis spectrophotometry. The stability constants for the binary and ternary complexes of norfloxacin were always higher than those found for ofloxacin and comparing the values obtained for the binary and ternary species (DeltalogK) it is possible to conclude that the ternary complexes are more stable than the binary ones, suggesting that an interaction occurs between the ligands in the ternary complexes. From the distribution diagrams it is possible to state that at physiological pH 7.4, the copper ternary complexes, are the main species in solution not only at the concentration used to determined the stability constants but also at the minimum inhibitory concentration. The antibacterial activity of these complexes, in different bacterial strains, was determined, at physiological pH, and the results obtain show that these ternary complexes may be good candidates as metalloantibiotics. PMID- 17118588 TI - Evaluation of bioadhesive potential and intestinal transport of pegylated poly(anhydride) nanoparticles. AB - Nanoparticles based on the poly(methyl vinyl ether-co-maleic anhydride) were pegylated with different types of PEGs, namely, two hydroxyl-functionalized PEGs (PEG and mPEG) and two amino-PEGs (DAE-PEG and DAP-PEG). The resulted nanoparticles demonstrated reduction of the negative surface charge compared to the non-modified particles. Further, in vivo experiments showed that all types of pegylated particles possessed higher affinity to adhere to intestinal rather than to the stomach mucosa. Higher bioadhesive potential was observed in the case of PEG-NP and DAE-PEG-NP which was attributed to the flexibility and specific properties of the surface "brush" layer of these particles. The lower bioadhesive potential of mPEG-NP was due to the low presence of coating "brush" layer, whereas for DAP-PEG-NP to the fact that the double end coupled chains ("loop" conformation) were not available for intensive interactions with the mucosa. The observations made by optic microscopy illustrated an intracellular transport of PEG-NP in vivo with preferable location in the apical area of enterocytes. PMID- 17118589 TI - Investigation on the photostability of tretinoin in creams. AB - In this investigation, the photodegradation of some tretinoin cream formulations was evaluated. Several oils were selected to prepare the cream formulations: olive oil, maize oil, castor oil, isopropyl myristate and Miglyol 812. A solubility study showed that tretinoin is best soluble in castor oil (0.60g/100ml), followed by isopropyl myristate, maize oil, Miglyol 812 and olive oil, respectively, 0.35, 0.30, 0.29 and 0.22g/100ml. The photostability of tretinoin in oils is comparable with the photostability of a tretinoin lotion (ethanol/propylene glycol 50/50), castor oil and olive oil giving slightly better results than the other oils. Investigation of the photodegradation of tretinoin in o/w creams, prepared with the same oils as mentioned above, revealed that tretinoin is far more stable in the cream formulations than in the respective oils, however it is not clear whether this is due to the formulation or due to a different irradiation technique. Tretinoin seemed to be most stable in the olive oil cream, followed by the castor oil cream. However microscopic investigation revealed the presence of tretinoin crystals in the olive oil cream, while the other creams were free of it. As a conclusion, one can say that the cream prepared with castor oil seems to be the most suitable one, in terms of solubility of tretinoin and in terms of photostability. PMID- 17118590 TI - Surface activity of a non-micelle forming compound containing a surface-active impurity. AB - PURPOSE: The purpose of this study is to characterize the surface activity of a water-soluble compound and its ability to form aggregates/micelles. METHODS: Aqueous solutions of the compound were prepared at various concentrations. Surface tension was determined using drop volume and Wilhelmy plate methods. Moreover, conductivity and osmolality measurements of aqueous solutions were also determined at various concentrations. RESULTS: Even though the compound appeared to be surface active, no change in the slope was found of either molar conductivity (Scm(2)/mol) versus square root of concentration (R(2)=0.994) or osmolality (mOsm/kg) versus concentration (R(2)=0.999). Moreover, no clear critical micelle concentration was observed when surface tension was plotted versus log concentration. These results indicated no micelle formation in these solutions. In order to investigate this behavior further, the main impurity in the compound was also tested. Surface tension measurements of solutions containing different concentrations of the impurity indicated that the impurity was more surface active than the compound. CONCLUSIONS: This study shows the importance of characterizing the behavior of surface-active compounds using multiple techniques. This work also emphasizes the importance of determining whether surface activity in aqueous solutions is due to the main compound, its impurities or both. PMID- 17118591 TI - Effects of solid-state reaction between paracetamol and cloperastine hydrochloride on the pharmaceutical properties of their preparations. AB - Tablets containing both paracetamol (PM) and cloperastine hydrochloride (CLH) in a combination formulation prepared by standard vertical granulation technology were found to have altered pharmaceutical properties. The hardness and disintegration time of tablets containing both PM and CLH gradually increased during storage, and the cross-screw did not operate smoothly during preparation of the mixed powder. The objective of the present study was to investigate the mechanism of formation of eutectic mixtures consisting of PM and CLH. Binary mixtures of PM and CLH in various proportions were prepared as physical mixtures and analyzed by DSC to study their thermal behavior. Phase diagrams obtained from the endothermic peaks due to melting of physical mixtures of PM and CLH demonstrated the formation of eutectic mixtures with eutectic temperatures of 86.9-110.2 degrees C depending on the ratio of constituents. The formation of the eutectic mixture was studied for a 50:50 mol.% ratio of PM and CLH. PXRD analysis revealed that the eutectic mixture of PM and CLH is structurally different from native PM and CLH. The most probable interaction sites between PM and CLH were demonstrated by DSC analysis of a binary mixture of PM and CLH prepared by melt quenching. PMID- 17118592 TI - Zootherapeutic practices among fishing communities in North and Northeast Brazil: a comparison. AB - This paper compares the medicinal uses of animals in fishing communities located in the North and Northeastern regions of Brazil. Data were obtained through field surveys conducted in the Northern state of Para and in the NE states of Paraiba, Piaui and Maranhao. We interviewed 137 people (67 men and 70 women), who provided information on animal species used as remedies, body parts used to prepare the remedies, and illnesses to which the remedies were prescribed. Comparisons were made using cluster analysis. Interviewees quoted 138 animal taxa, 11 of which are listed in the Brazilian list of threatened species. Animals were used to treat 100 illnesses. Results suggest that similarities in the repertoire of medicinal resources chosen by the surveyed communities reflects both a geographic/cultural continuum and the local accessibility/availability of the resources. PMID- 17118593 TI - Clematis vitalba L. aerial part exhibits potent anti-inflammatory, antinociceptive and antipyretic effects. AB - Extracts obtained from the dried aerial parts of Clematis species are used as folk remedy worldwide for the treatment of various inflammatory ailments such as rheumatism and to reduce fever. In order to test the effectiveness of extracts, fractions and subfractions from dried Clematis vitalba L. (Ranunculaceae) aerial parts were studied on mice. Extracts are shown to have a potent effect on carrageenan-induced hind paw edema and acetic acid-induced increased vascular permeability models. Through bioassay-guided fractionation procedures a new C glycosylflavon, 4'-O-coumaroyl-isovitexine (vitalboside) was isolated as the main active ingredient of the aerial parts. Vitalboside showed a potent and dose dependent (in 75 and 150 mg/kg does, per os) in vivo anti-inflammatory activity against acute (carrageenan-, serotonin- and PGE(2)-induced hind paw edema model, castor oil-induced diarrhea), subacute (subcutaneous air-pouch) and chronic (Freund's complete adjuvant-induced arthritis) models of inflammation. The same compound was also isolated as the main antinociceptive principle which was assessed by using the models based on the inhibition of p-benzoquinone-induced writhings, as well as antipyretic activity against Freund's complete adjuvant induced increased body temperature. Acute and subchronic toxicity studies were also performed. PMID- 17118594 TI - Interpretation of ante-mortem stature estimates in South Africans. AB - When human skeletal remains of unknown individuals are analysed, the estimation of ante-mortem stature forms a key part of the report. However, for many people it may be difficult to judge the height of their missing relative in metric values. In this paper, we report on the statures of 2052 black, 483 coloured and 880 white South Africans of both sexes, measured during the last few years. It is shown that the statures of whites are significantly higher than those of the other two groups. The statures of the black and coloured groups are similar. It is proposed that the stature estimates provided in forensic reports are interpreted as tall, medium or short, based on whether they fall in the upper 25%, middle 50% or lower 25% of the distribution for the specific population. PMID- 17118595 TI - Investigation into dizziness before and after Epley's maneuver for benign paroxysmal positional vertigo using stabilometry. AB - OBJECTIVE: To investigate the cause of dizziness after Epley's maneuver using stabilometry. METHODS: Subjects were 35 patients with posterior canal type BPPV. First we performed stabilometry, next Epley's maneuver, then stabilometry was performed again. The enveloped area and the locus length per second of stabilometry were compared. RESULTS: Seventeen patients reported a feeling of dizziness after the treatment, whereas the other 18 patients did not. The patients were divided into two groups: 17 patients with dizziness (WD) and 18 patients without dizziness (WOD) after the treatment. In the WD group, the enveloped area (P=0.0495) and the locus length per second (P=0.0099) before Epley's maneuver were significantly lower than those after Epley's maneuver. In the WOD group, there was no significant difference in the stabilometry parameter between values obtained before and after Epley's maneuver. There were no significant differences found in either parameter between these two groups either before or after applying Epley's maneuver. CONCLUSIONS: Epley's maneuver is a treatment for the posterior semicircular canal. Malfunction of the otolith organ remains. It is considered that the cause of dizziness after Epley's maneuver comes from otolith dysfunction. PMID- 17118596 TI - Case report: Meniere's disease and otosclerosis--different outcomes of the same disease? AB - The etiologies of Meniere's disease and otosclerosis are largely unknown. An association between these two diseases has been proposed on both a clinical and histopathologic basis but the causal relationship is controversial. In this paper we report two families in which both otosclerosis and Meniere's disease are inherited as independent phenotypes suggesting that the two diseases represent different outcomes of the same mutation. Thus the Meniere's disease occasionally seen in otosclerotic patients might not be caused by otosclerosis, but rather by a molecular defect leading to endolymphatic hydrops and/or clinical otosclerosis. PMID- 17118597 TI - Sulpiride plus hydroxyzine decrease tinnitus perception. AB - OBJECTIVES: The aim of the study is to confirm the effectiveness of sulpiride and hydroxyzine in tinnitus patients. The administration of sulpiride, a D2 antagonist of dopamine receptors, together with hydroxyzine, a subcortical sedative, covers the areas of tinnitus perception. METHODS: A prospective, randomized, single blinded, placebo-control study was done in general otorhinolaryngology consultations for 2002-2004 in Seville and Zaragoza (Spain). One hundred and fifty patients consulted for subjective tinnitus. They were included randomly in three groups of 50. A group took sulpiride (50 mg/8 h) alone, other the same dose of sulpiride plus hydroxyzine (25 mg/12 h), and the third placebo (lactose), for 1 month. One hundred and twenty-two patients completed the study. Clinical history, tonal audiometry, tympanometry, and tinnitometry were done in the beginning and end of the study. Subjective Grading of Tinnitus Perception and visual analogical scale (0-10) were done for result evaluation. RESULTS: Based on the Subjective Grading of Tinnitus Perception, tinnitus perception diminished by 56% in patients treated with sulpiride and by 81% in patients treated with sulpiride plus hydroxyzine. Based on the visual analogical scale, tinnitus perception diminished from 7.8 to 6.3 in the patients treated with sulpiride, and from 7.8 to 5.1 in those treated with sulpiride plus hydroxyzine. CONCLUSIONS: Sulpiride plus hydroxyzine decreases tinnitus perception. Tinnitus auditolimbic dopaminergic pathway opens wide therapeutical implications. PMID- 17118598 TI - A cross-national study of transitions in deficit counts in two birth cohorts: implications for modeling ageing. AB - Generally, health does not improve with age, and many physical and physiological functions are known to decline. These changes do not occur uniformly, however; for many reasons, some people experience significant improvement in their health over non-trivial time intervals. Earlier, we showed that 5-year transitions in health status in elderly people (age 65+ years) can be modeled as a stochastic process, using a modified Poisson distribution with four readily interpretable parameters. The original description was based on follow-up of a single cross sectional study, thus mixing age and cohort effects. Here, we again used a multistate Markov chain to model 5-year deficit accumulation in relation to frailty in both a Swedish birth cohort (aged 70 years at inception) and, from the original cross-sectional study, a Canadian birth cohort, aged 69-71. In both datasets, we found again that a modified Poisson describes the transition in health status with high precision. The parameters of the model though different, are close to each other, even though the cohorts are from different countries, were assembled 20 years apart, and counted different deficits. The model suggests that all health transitions, including health improvement, worsening, and death, can be summarized in a unified stochastic model with a few interpretable parameters. PMID- 17118599 TI - Evaluation of sex differences on mitochondrial bioenergetics and apoptosis in mice. AB - It has been postulated that the differences in longevity observed between organisms of different sexes within a species can be attributed to differences in oxidative stress. It is generally accepted that differences are due to the higher female estrogen levels. However, in some species males live the same or longer despite their lower estrogen values. Therefore, in the present study, we analyze key parameters of mitochondrial bioenergetics, oxidative stress and apoptosis in the B6 (C57Bl/6J) mouse strain. There are no differences in longevity between males and females in this mouse strain, although estrogen levels are higher in females. We did not find any differences in heart, skeletal muscle and liver mitochondrial oxygen consumption (State 3 and State 4) and ATP content between male and female mice. Moreover, mitochondrial H(2)O(2) generation and oxidative stress levels determined by cytosolic protein carbonyls and concentration of 8 hydroxy-2'-deoxyguanosine in mitochondrial DNA were similar in both sexes. In addition, markers of apoptosis (caspase-3, caspase-9 and mono- and oligonucleosomes: the apoptosis index) were not different between male and female mice. These data show that there are no differences in mitochondrial bioenergetics, oxidative stress and apoptosis due to gender in this mouse strain according with the lack of differences in longevity. These results support the Mitochondrial Free Radical Theory of Aging, and indicate that oxidative stress generation independent of estrogen levels determines aging rate. PMID- 17118600 TI - A search for principles of disability using experimental impairment of Drosophila melanogaster. AB - The results of life table experiments to determine the effects of artificial impairment (leg amputation) in 7500 Drosophila melanogaster adults revealed that the extent to which life expectancy was reduced in impaired individuals was conditional on: (1) leg location and number amputated--front leg had greatest impact and the number of legs amputated directly correlated with mortality impact; (2) age of amputation--the greatest relative reduction in remaining life expectancy occurred when young flies were impaired; (3) vial orientation- mortality in impaired flies was the least when vials held upside-down (most friendly environment) and the greatest when they were right-side up (least friendly environment); and (4) sex--male mortality was increased more than female mortality in nearly all impairment treatments. These results were used to formulate a set of general principles of disability that would apply not only to humans but to all organisms. PMID- 17118601 TI - Immunity and ageing in man. AB - Immunosenescence resulting in decreased ability to control infectious disease contributes to morbidity and mortality not only in the very elderly, but in all likelihood already from middle age. Studying immunity in humans is therefore essential for developing treatments to restore dysregulated immune responses and assure healthy longevity. The past year has seen many significant advances in our knowledge of age-associated alterations to immunity in elderly people, only some of which can be briefly reviewed here. PMID- 17118602 TI - Aging affects passive stiffness and spindle function of the rat soleus muscle. AB - Aging affects many motor functions, notably the spinal stretch reflexes and muscle spindle sensitivity. Spindle activation also depends on the elastic properties of the structures linked to the proprioceptive receptors. We have calculated a spindle efficacy index, SEI, for old rats. This index relates the spindle sensitivity, deduced from electroneurograms recording (ENG), to the passive stiffness of the muscle. Spindle sensitivity and passive incremental stiffness were calculated during ramp and hold stretches imposed on pseudo isolated soleus muscles of control rats (aged 4 months, n=12) and old rats (aged 24 months, n=16). SEI were calculated for the dynamic and static phases of ramp (1-80 mm/s) and for hold (0.5-2mm) stretches imposed at two reference lengths: length threshold for spindle afferents discharges, L(n) (neurogram length) and slack length, L(s). The passive incremental stiffness was calculated from the peak and steady values of passive tension, measured under the stretch conditions used for the ENG recordings, and taking into account the muscle cross-sectional area. The pseudo-isolated soleus muscles were also stretched to establish the stress-strain relationship and to calculate muscle stiffness constant. The contralateral muscle was used to count muscle spindles and spindle fibers (ATPase staining) and immunostained to identify MyHC isoforms. L(n) and L(s) lengths were not significantly different in the control group, while L(n) was significantly greater than L(s) in old muscles. Under dynamic conditions, the SEI of old muscles was the same as in controls at L(s), but it was significantly lower than in controls at L(n) due to increased passive incremental stiffness under the stretch conditions used to analyze the ENG. Under static conditions, the SEI of old muscles was significantly lower than control values at all the stretch amplitudes and threshold lengths tested, due to increased passive incremental stiffness and decreased spindle sensitivity at L(s). The muscle stiffness constant values were greater in old muscles than in controls, confirming the changes in elastic properties under passive conditions due to aging. Aging also altered the intrafusal fibers: it increased the mean number of intrafusal fibers and the contents in the slow, neonatal and developmental isoforms intrafusal of MyHC have been modified. These structural modifications do not seem great enough to counteract the loss of the spindle sensitivity or the spindle efficacy under passive conditions and after the nerve was severed. However, they may help to maintain the spindle afferent message under natural conditions and under fusimotor control. PMID- 17118603 TI - Imaging of hepatic steatosis and fatty sparing. AB - Radiology has gained importance in the non-invasive diagnosis of hepatic steatosis. Ultrasonography is usually the first imaging modality for the evaluation of hepatic steatosis. Unenhanced CT with or without dual kVp measurement and MRI with in and out of phase sequence can allow objective evaluation of hepatic steatosis. However, none of the imaging modalities can differentiate non-alcoholic steatohepatitis/fatty liver disease from simple steatosis. Evaluation of hepatic steatosis is important in donor evaluation before orthotopic liver transplantation and hepatic surgery. Recently, one-stop shop evaluation of potential liver donors has become possible by CT and MRI integrating vascular, parenchymal, volume and steatosis evaluation. Moreover hepatic steatosis (diffuse, multinodular, focal, subcortical, perilesional, intralesional, periportal and perivenular), hypersteatosis and sparing (geographic, nodular and perilesional or peritumoral) can cause diagnostic problems as a pseudotumor particularly in the evaluation of oncology patients. Liver MRI is used as a problem-solving tool in these patients. In this review, we discuss the current role of radiology in diagnosing, quantifying hepatic steatosis and solutions for diagnostic problems associated with fatty infiltration and sparing. PMID- 17118604 TI - A rapid, simple, specific liquid chromatographic-electrospray mass spectrometry method for the determination of finasteride in human plasma and its application to pharmacokinetic study. AB - A fast, accurate, sensitive, selective and reliable method using reversed-phase high-performance liquid chromatography-mass spectrometry coupling with an electrospray ionization interface was developed and validated for the determination of finasteride in human plasma. After deprotienation with acetonitrile, centrifugation, evaporation to dryness and dissolving in mobile phase, satisfactory separation was achieved on a Hypersil-Keystone C(18) reversed phase column using a mobile phase consisting of acetonitrile-water (46:54, v/v), 0.1% acetic acid and 0.1% trifluoracetic acid. Carbamazepine (IS) was used as internal standard. This method involved the use of the [M+H](+) ions of finasteride and IS at m/z 373 and 237 with the selective ion monitoring (SIM) mode. The calibration curve was linear in the range of 0.2-120 ng ml(-1). The limit of quantification for finasteride in plasma was 0.2 ng ml(-1) with good accuracy and precision. The intra-assay precision and accuracy were in the range of 2.1-11.2% and -1.3% to 8.5%, respectively. The inter-assay precision and accuracy were in the order of 3.4-12.1% and -1.5% to 11.5%, respectively. The mean sample extract recoveries of the method were higher than 85% and 74% for finasteride and internal standard (IS), respectively. The assay has been successfully used to estimate the pharmacokinetics of finasteride after oral administration of a 5mg tablet of finasteride to 24 healthy volunteers. PMID- 17118605 TI - Optimization of homonuclear 2D NMR for fast quantitative analysis: application to tropine-nortropine mixtures. AB - Quantitative analysis by (1)H NMR is often hampered by heavily overlapping signals that may occur for complex mixtures, especially those containing similar compounds. Bidimensional homonuclear NMR spectroscopy can overcome this difficulty. A thorough review of acquisition and post-processing parameters was carried out to obtain accurate and precise, quantitative 2D J-resolved and DQF COSY spectra in a much reduced time, thus limiting the spectrometer instabilities in the course of time. The number of t(1) increments was reduced as much as possible, and standard deviation was improved by optimization of spectral width, number of transients, phase cycling and apodization function. Localized polynomial baseline corrections were applied to the relevant chemical shift areas. Our method was applied to tropine-nortropine mixtures. Quantitative J resolved spectra were obtained in less than 3 min and quantitative DQF-COSY spectra in 12 min, with an accuracy of 3% for J-spectroscopy and 2% for DQF-COSY, and a standard deviation smaller than 1%. PMID- 17118606 TI - Investigation of the metabolism of substance P at the blood-brain barrier using LC-MS/MS. AB - Substance P (SP) has been associated with pain and depression as well as neurodegenerative diseases. Many of these diverse actions of SP can potentially be attributed to SP metabolites generated at the blood-brain barrier (BBB). In this study, the metabolism of SP was investigated using an in vitro model of the BBB and LC-MS/MS. Substance P metabolism was found to be non-saturable in the concentration range of 100 nM to 10 microM, with approximately 70% of the peptide remaining intact after 5 h. The major metabolites of SP were identified by MS as 3-11 and 5-11. Two previously unreported metabolites, 5-11 and 6-11, were also found in our studies. Several additional minor SP metabolites, including 1-9 and 2-11, were also identified. A profile of the SP metabolites generated by the BBB over time was obtained. The results from the present study provide a better understanding of the role of the blood-brain barrier in the pharmacology of SP. PMID- 17118607 TI - Kinetics and mechanism of degradation of lithospermic acid B in aqueous solution. AB - The degradation of lithospermic acid B (LAB) was investigated as a function of buffer concentration, pH and temperature. Stability tests were performed using a stability-indicating high-performance liquid chromatography (HPLC) with UV-vis detection. The degradation followed pseudo-first-order kinetics under all experimental conditions. The maximum stability of LAB was observed at pH 2.0. The logk(pH)-pH profile described by specific acid-base catalysis and water molecules agreed with the experimental results. The overall degradation rate constant as a function of the temperature under the given conditions obeyed the Arrhenius equation. The chemical fate of LAB in mild acidic solution was investigated, and nine degradation products were detected and tentatively identified by LC-MS analysis. The primary degradation pathway involving the cleavage of ester bond and ring-opened of benzofuran in the LAB are proposed. PMID- 17118608 TI - Complexation study of cinalukast and montelukast with cyclodextrines. AB - A fluorimetric study on the spectral characteristics of two antileukotrienes, cinalukast and montelukast, has been performed. Ionization constants of both of them have been photometrically calculated. Cinalukast pK(a) in ethanol:water 50:50 (v/v) medium resulted to be 2.2+/-0.1. Because the spectral characteristics of montelukast are widely affected by the solvent nature, pK(a) was estimated in two different ethanol:water media, 70:30 (v/v) and 10:90 (v/v) and the values calculated were pK(a)=2.9+/-0.1, and pK(a1)=2.0+/-0.1 and pK(a2)=6.5+/-0.1, respectively. It has been proven that the fluorescence of both, cinalukast and montelukast, is significantly intensified in the presence of cyclodextrins (CyDs). The host-guest complexation processes between cinalukast and alpha-CyD or heptakis-(2,6-di-O-methyl)-beta-cyclodextrin (DIMEB) and between montelukast and DIMEB have been investigated by fluorescence spectroscopy. A 1:1 stoichiometric ratio was established for the three studied inclusion complexes. The changes produced on the fluorescence of cinalukast or montelukast, when they are included on the hydrophobic CyD cavity are used to calculate their association constants by a non-linear regression method. Semiempirical MO calculations using AM1 method were performed in order to characterize the studied inclusion complexes. A new method for cinalukast determination in human serum, based on the fluorescence of the complex cinalukast-DIMEB exhibiting limit of detection of 7.95 ng mL(-1) has been proposed with satisfactory results. Adequate recovery values between 95 and 103% were calculated at five different concentration levels. PMID- 17118609 TI - Quality control of Chinese medicinal preparations LC/ESI(+)/MS/MS analyses of saikosaponins-a and -c as markers of Bupleuri radix samples. AB - We have used LC/ion trap tandem MS analysis to determine saikosaponin-a and -c as target markers in crude 70% methanol extracts from three different species of Bupleuri radix and the 10 most-popular Chinese medicinal preparations containing "Chaihu" (B. radix) without any clean-up. The optimal ionization characteristics were obtained when using positive-ion electrospray ionization (ESI) with 50 microM sodium acetate as an additive in the mobile phase. We observed good linearity over the range from 0.02 to 2 microg/ml for saikosaponin-a and from 0.02 to 1 microg/ml for saikosaponin-c. The intra-day precisions varied between 3.3 and 8.8% for saikosaponin-a and 0.3 and 11.1% for saikosaponin-c. The limits of detection were 0.01 microg/ml for both markers. The recoveries of saikosaponin a and -c from the extract of a medicinal preparation sample (Chai-Hu-Ching-Gan Tang, No. 13 in the table of section Analysis on actual samples) were 97 and 100%, respectively, at a 1 microg/ml spiking concentration of each marker. The highest concentrations of saikosaponin-a and -c among the three B. radixes were found in B. kaoi Liu Chao & Chuang (10.1 mg/g) and in B. falcatum (3.4 mg/g), respectively. The amounts of these saikosaponins in the 10 Chinese medicinal preparations ranged between 0.11 and 1.22 mg/g for saikosaponin-a and between 0.01 and 0.33 mg/g for saikosaponin-c. PMID- 17118610 TI - LC and LC-MS study of stress decomposition behaviour of isoniazid and establishment of validated stability-indicating assay method. AB - Isoniazid was subjected to different ICH prescribed stress conditions of thermal stress, hydrolysis, oxidation and photolysis. The drug was stable to dry heat (50 and 60 degrees C). It showed extensive decomposition under hydrolytic conditions, while it was only moderately sensitive to oxidation stress. The solid drug turned intense yellow on exposure to light under accelerated conditions of temperature (40 degrees C) and humidity (75% RH). In total, three major degradation products were detected by LC. For establishment of stability-indicating assay, the reaction solutions in which different degradation products were formed were mixed, and the separation was optimized by varying the LC conditions. An acceptable separation was achieved using a C-18 column and a mobile phase comprising of water:acetonitrile (96:4, v/v), with flow rate and detection wavelength being 0.5 ml min(-1) and 254 nm, respectively. The degradation products appeared at relative retention times (RR(T)) of 0.71, 1.34 and 4.22. The validation studies established a linear response of the drug at concentrations between 50 and 1000 microg ml(-1). The mean values (+/-R.S.D.) of slope, intercept and correlation coefficient were 35,199 (+/-0.88), 114,310 (+/-4.70) and 0.9998 (+/-0.01), respectively. The mean R.S.D. values for intra- and inter day precision were 0.24 and 0.90, respectively. The recovery of the drug ranged between 99.42 and 100.58%, when it was spiked to a mixture of solutions in which sufficient degradation was observed. The specificity was established through peak purity testing using a photodiode array detector. The method worked well on application to marketed formulation of isoniazid, and a fixed-dose combination containing isoniazid and ethambutol HCl. It was even extendable to LC-MS studies, which were carried out to identify the three degradation products. The m/z values of the peaks at RR(T) 0.71 and RR(T) 1.34 matched with isonicotinic acid and isonicotinamide, respectively. The product appearing at RR(T) 4.22 was isolated using preparative LC-MS, and turned out to be a yellow compound that was identified as isonicotinic acid N'-(pyridyl-4-carbonyl)-hydrazide based on mass, FTIR and (1)H/(13)C NMR spectral data. The same was indicated to be responsible for discolouration of isoniazid bulk drug substance and formulations, which is a familiar problem. The mechanism of formation of the said compound is outlined. PMID- 17118611 TI - Validation of an ELISA for determination of antibodies induced in monkeys against Epi-hNE4, a recombinant protein inhibitor of human neutrophil elastase. AB - The engineered protein inhibitor of human neutrophil elastase, Epi-hNE4, is being developed for the treatment of cystic fibrosis. Like many recombinant proteins, Epi-hNE4 may induce antibodies in pre-clinical species and in humans. The aim of this report was to validate an ELISA to assess its immunogenicity in monkeys. We have designed and optimized a classical ELISA in which Epi-hNE4 was coated directly on microtitre plates and the antibodies were detected using a secondary antibody labelled with peroxidase. We report implementation of the recent recommendations proposed for the validation of immunogenicity assessment. The cut off point was determined by means of statistical analysis of negative samples. Linearity, reproducibility, stability and specificity were estimated using quality control samples obtained from a pool of positive samples. The method was applied to monkeys given Epi-hNE4 by inhalation. A confirmation test and a neutralization assay were developed in order to further assess positive samples. In conclusion, we present here one of the first examples of validation in application of recent recommendations [A.R. Mire-Sluis, Y.C. Barrett, V. Devanarayan, E. Koren, H. Liu, M. Maia, T. Parish, G. Scott, G. Shankar, E. Shores, S.J. Swanson, G. Taniguchi, D. Wierda, L.A. Zuckerman, J. Immunol. Methods 289 (2004) 1-16]. PMID- 17118612 TI - Assessment of the immunogenicity of different interferon beta-1a formulations using ex vivo T-cell assays. AB - Recombinant homologues of human proteins have the potential to induce an immunogenic response when used therapeutically. Each of the three interferon (IFN) beta therapies currently approved for multiple sclerosis can induce the development of neutralising antibodies, the full effects of which on IFN beta therapy remain unclear. To investigate the immunogenicity of the currently licensed formulation of subcutaneous IFN beta-1a, 22 or 44 microg three times weekly (Rebif), two new formulations of IFN beta-1a (Rebif) New Formulation [RNF]1 and RNF2) have been developed. In this study, the immunogenicity of the current formulation was investigated against RNF1, RNF2 and an IFN beta standard using ex vivo T-cells. Dendritic cells, isolated from peripheral blood monocytes donated by 26 healthy volunteers, were matured in vitro and incubated with the test antigens for 6h. Autologous CD4(+) T-cells from the same donors were added and further incubated before cytokine release was assessed by ELISpot assay and proliferation by [(3)H]thymidine pulse. Secretion of T-cell-derived interleukin-2 was 79%, 66% and 105% in incubations with RNF1, RNF2 and the current formulation, respectively (normalised to secretion with the IFN standard; p<0.05, RNF2 versus the current formulation). Secretion of IFN gamma was highly variable between donors, with no significant difference observed between formulations. Normalised values for T-cell proliferation were 56%, 44% and 88% with RNF1, RNF2 and the current formulation, respectively (p<0.05, RNF2 versus the current formulation). The immunogenic potential of RNF2 is significantly lower than that of the current formulation when tested ex vivo. This result is now being confirmed through ongoing clinical trials. PMID- 17118613 TI - Simultaneous determination of p-aminobenzoic acid and its metabolites in the urine of volunteers, treated with p-aminobenzoic acid sunscreen formulation. AB - p-Aminobenzoic acid (PABA) and its metabolites (p-aminohippuric acid, p acetamidobenzoic acid, and p-acetamidohippuric acid) were detected using high performance liquid chromatography with an electrochemical (carbon paste) detector (HPLC-ECD). For direct current (dc) mode, with the current at a constant potential, and measurements with suitable experimental parameters, a linear concentration from 0.125 to 1.80 microg/ml was found. The detection limit was approximately 2.0 ng/ml. A carbon paste coulometric detector was used to demonstrate that PABA and its metabolites are electrochemically oxidized in acidic media, and to determine, by analyzing human urine, the percutaneous absorption of PABA and its metabolites. Findings using HPLC-ECD and HPLC with an ultraviolet detector (HPLC-UV) were comparable. PMID- 17118614 TI - Validated high-throughput HPLC assay for nimesulide using a short monolithic column. AB - High samples analysis rate is a key demand in modern pharmaceutical analysis, especially during new product development and validation of industrial-scale manufacturing process. The present study reports a validated HPLC assay for the dissolution studies of nimesulide-containing tablets (Lizepat 100 mg/tab, Cosmopharm Ltd., Korinthos, Greece). Using a 50 mm x 4.6 mm i.d. monolithic column (Chromolith, Merck) and acetonitrile-phosphate buffer (pH 7.0; 10 mM) (34:66, v/v) as the mobile phase, the separation cycle was completed in 60s at a flow rate of 4.0 ml min(-1). The assay was validated in terms of selectivity against potential impurities of the active ingredient, detection and quantification limits, linearity, accuracy and inter-/intra-day precision. Results from the application of the HPLC method to the accelerated and long-term dissolution stability control of Lizepat tablets (Lot 005) are reported. PMID- 17118615 TI - Quantitative determination and sampling of azathioprine residues for cleaning validation in production area. AB - Cleaning validation is an integral part of current good manufacturing practices in any pharmaceutical industry. Nowadays, azathioprine and several other pharmacologically potent pharmaceuticals are manufactured in same production area. Carefully designed cleaning validation and its evaluation can ensure that residues of azathioprine will not carry over and cross contaminate the subsequent product. The aim of this study was to validate simple analytical method for verification of residual azathioprine in equipments used in the production area and to confirm efficiency of cleaning procedure. The HPLC method was validated on a LC system using Nova-Pak C18 (3.9 mm x 150 mm, 4 microm) and methanol-water acetic acid (20:80:1, v/v/v) as mobile phase at a flow rate of 1.0 mL min(-1). UV detection was made at 280 nm. The calibration curve was linear over a concentration range from 2.0 to 22.0 microg mL(-1) with a correlation coefficient of 0.9998. The detection limit (DL) and quantitation limit (QL) were 0.09 and 0.29 microg mL(-1), respectively. The intra-day and inter-day precision expressed as relative standard deviation (R.S.D.) were below 2.0%. The mean recovery of method was 99.19%. The mean extraction-recovery from manufacturing equipments was 83.5%. The developed UV spectrophotometric method could only be used as limit method to qualify or reject cleaning procedure in production area. Nevertheless, the simplicity of spectrophotometric method makes it useful for routine analysis of azathioprine residues on cleaned surface and as an alternative to proposed HPLC method. PMID- 17118617 TI - Literacy, self-efficacy, and HIV medication adherence. AB - OBJECTIVE: We examined the relationship between patient literacy level and self reported HIV medication adherence, while estimating the mediating roles of treatment knowledge and self-efficacy on this relationship. METHODS: Structured patient interviews with a literacy assessment, supplemented by medical chart review, were conducted among 204 consecutive patients receiving care at infectious disease clinics in Shreveport, Louisiana and Chicago, Illinois. Literacy was measured using the Rapid Estimate of Adult Literacy in Medicine (REALM), while the Patient Medication Adherence Questionnaire (PMAQ) was used to assess medication self-efficacy and adherence to antiretroviral regimens in the past 4 days. RESULTS: Approximately one-third of patients (30.4%) were less than 100% adherent to their regimen, and 31.4% had marginal to low literacy skills. In multivariate analyses, low literate patients were 3.3 times more likely to be non adherent to their antiretroviral regimen (p < 0.001). Patients' self-efficacy, but not knowledge, mediated the impact of low literacy on medication adherence (AOR 7.4, 95% CI 2.7-12.5). CONCLUSION: While low literacy was a significant risk factor for improper adherence to HIV medication regimens in our study, self efficacy mediated this relationship. PRACTICE IMPLICATIONS: Comprehensive intervention strategies that go beyond knowledge transfer may be needed to address self-efficacy among patients across all literacy levels to be successful in the management of difficult medication schedules. PMID- 17118616 TI - Factors associated with medication hassles experienced by family caregivers of older adults. AB - OBJECTIVE: We wished to identify potential factors associated with medication administration hassles, daily irritants, among informal caregivers who provide long-term medication assistance to persons aged 55 or older. METHODS: A sample of 156 informal caregivers were recruited from seven states and several types of settings. The dependent variable was scores on the Family Caregiver Medication Administration Hassles Scale (FCMAHS). Independent variables included in the analyses were medication complexity; caregiver's gender, ethnicity, relationship to recipient, length of time in caregiving, education, and employment outside the home; care recipient's physical capacity and mental capacity; and whether the caregiver and care recipient live together. After preliminary analysis to reduce the number of independent variables, the remaining variables were included in a linear model (GLM procedure). Possible interactions and residuals were considered. RESULTS: Whites and Hispanics experience greater medication administration hassles than other groups, and perceived hassle intensity increases with medication complexity. Medication administration hassle scores increase with increasing education levels up to a high school degree, after which they remain consistently high. Caregivers whose care recipients have moderate levels of cognitive functioning have higher medication administration hassles scores than those whose care recipients have very high or very low cognitive functioning. CONCLUSION: The preliminary set of significant variables can be used to identify caregivers who may be at risk of experiencing medication administration hassles, increased stress, and potentially harmful events for their care recipients. PRACTICE IMPLICATIONS: Family caregivers are accepting complex caregiving responsibility for family members while receiving little or no support or assistance with caregiving hassles associated with this duty. The FCMAHS offers the means to monitor how caregivers are handling the daily irritants involved with medication administration so that educational interventions can be provided before hassles lead to more serious stress and strain. PMID- 17118618 TI - Mammary arteriovenous differences of glucose, insulin, prolactin and IGF-I in lactating sows under different protein intake levels. AB - Mammary uptake of nutrients is dependent on their availability in the circulation but the role of hormones in that process is not known. Arteriovenous differences (AVD) of glucose and key hormones across the mammary glands were therefore determined in sows fed varying levels of protein. Sixteen lactating sows (four/dietary treatment) were fed a 7.8, 13.0, 18.2 or 23.5% crude protein (CP) isocaloric diet throughout lactation and their litters were standardized to 11 pigs within 48 h of birth. The anterior main mammary vein and a carotid artery were cannulated on day 4+/-1 of lactation and blood samples were collected every 30 min over 6h on days 10, 14, 18 and 22 of lactation to measure glucose, insulin, IGF-I, and prolactin (PRL) concentrations. Amino acid data from these sows were previously published and used here to determine residual correlations. Dietary treatments had no effect on any of the insulin or PRL variables measured (P>0.1) and, on day 18 only, IGF-I AVD was greater (P=0.05) for sows on the 23.5% compared to the 18.2% diet. On days 18 and 22, sows fed the 13% CP diet had greater arterial, venous and AVD glucose concentrations than sows fed other diets (P<0.05). Total arterial amino acid concentrations were correlated to arterial insulin (P<0.001) and PRL (P<0.05) concentrations, but not to those of IGF-I (P>0.1). Mammary AVD for total (P<0.001) and essential amino acids (P<0.05) were correlated to arterial concentrations of insulin, but not to those of IGF-I (P>0.1) or PRL (P>0.1). Mammary AVD of both total (P<0.01) and essential (P<0.05) amino acids were also correlated to mammary PRL AVD. In conclusion, dietary protein level did not affect mammary AVD and circulating lactogenic hormone concentrations. Yet, amino acid utilization by the sow mammary gland seems to be regulated via both circulating insulin concentrations and PRL binding to and uptake by porcine mammary cells. PMID- 17118619 TI - Interaction of GH polymorphism with body weight and endocrine functions in Japanese black calves. AB - We assessed the interaction of GH gene polymorphisms (AA, AB and BB genotypes) with body weight and measures of endocrine function in Japanese black calves at 10 months of age. The average body weight for the BB genotype (281+/-5 kg) was significantly lower (P=0.0017, ANOVA) than those for the AA (324+/-9 kg) and AB (317+/-7 kg) genotypes. Plasma concentrations of insulin and IGF-I were greater for the AA genotype than for the AB genotype, and AB and BB genotypes, respectively. There were significant differences in the triglyceride and cholesterol concentrations among the GH genotypes. The area under the basal GH concentration was significantly greater (P=0.0314) for the AA genotype than for the two other genotypes. The incremental area over the basal GH concentrations in response to intravenous GHRH injection (0.4 microg/kg BW) was significantly smaller (P=0.0005) for the BB genotype than for the two other genotypes. In addition, linear regression analysis between GH incremental area induced by GHRH and body weight demonstrated that there was a positive linear correlation (r=0.6496, P<0.002) for incremental areas less than 600 ng min/ml, but a negative correlation (r=0.6473, P<0.05) for incremental areas over 600 ng min/ml. These findings indicate that the GH genotypes of the animals could be associated with difference in the GH response in Japanese black cattle at 10 months of age. We also observed a relationship between genotype and animal performances, but other studies on more animals in different conditions must be realized to make a definite conclusion. PMID- 17118620 TI - [Recommendations for good clinical practice concerning the application of Law No 2005-370 of April 2005 concerning the rights of patients at the end of life]]. PMID- 17118621 TI - New thymidylate synthase inhibitors induce apoptosis in melanoma cell lines. AB - Malignant melanoma is particularly resistant to conventional chemotherapy and radiotherapy. For this reason in the past years a huge variety of new compounds has been developed with potential chemotherapeutic activity which needs to be tested in vitro and in vivo. We investigated the in vitro action of three new experimental antifolate substances (MR7, MR21 and MR36) with a critical target for thymidylate synthase (TS), an essential enzyme for DNA synthesis. The response of two melanoma cell lines (SK-MEL-2 derived from malignant melanoma metastasis and SK-MEL-28 derived from primary malignant melanoma) was examined after treatment with these substances. The antifolate agents induced apoptosis in SK-MEL-2 and SK-MEL-28 cells as confirmed by the TUNEL technique and Comet Assay. Western-blot analysis showed a down-regulation of Bcl-2 protein level and PARP cleavage, otherwise p53 and Bax expressions were not modulated. Moreover, these antifolate-induced apoptosis was accompanied by both pro-caspase-9 and -8 activations. These results were supported by the use of the pan-caspases inhibitor Z-VAD-FMK that almost completely decreased the amount of apoptosis in both the melanoma cell lines treated with antifolate. In conclusion our results show that TS inhibitors are able to induce apoptosis through a caspase-mediated pathway, but without the involvement of the p53/Bax signalling. PMID- 17118622 TI - Phenotypic alterations and cytokine production in THP-1 cells in response to allergens. AB - In the induction phase of allergic contact hypersensitivity, dendritic cells (DCs), including Langerhans cells (LCs) present in epidermis, can trigger an efficient T cell response once they have matured in response to an allergen. Upon maturation, DCs have been shown to induce expression of several surface molecules and the up-regulation of cytokine production. We have previously shown that THP-1 cells, human acute monocytic leukemia cell line, can discriminate between allergens and irritants by measuring expression of surface markers, CD86 and CD54, following chemical exposure. At the same time, we have also reported that augmented expression of HLA and CD80, and production of IL-1beta were up regulated in THP-1 cells when treated with an allergen, 2,4-dinitrochlorobenzene (DNCB). In the present study, we first evaluated whether THP-1 cells induced the phenotypic changes and the production of cytokines, which are observed in the process of DC maturation, when treated with two known allergens, DNCB and nickel sulfate (NiSO(4)), and one irritant (sodium lauryl sulfate (SLS)). Exposure to DNCB and NiSO(4) induced significant augmentation of CD40 and CD83 expression as well as CD86 and CD54. Also, TNF-alpha and IL-8 secretion were markedly induced by DNCB and NiSO(4) in a dose-dependent manner. In addition, DNCB and NiSO(4) augmented CD1a expression and production of IL-6, respectively. On the contrary, SLS did not change any of these markers. We then evaluated a series of chemicals, including six known allergens (e.g., hydroquinone (HQ)) and two non-allergens (e.g., methyl paraben (MP)), in order to investigate the potential increase of CD86, CD54, CD40, and CD83 expression on THP-1 cells, and production of TNF-alpha and IL-8. Indeed, all tested allergens, except eugenol (EU), caused significant increased changes in at least four of the analyzed six markers, while non allergens did not induce any changes. EU significantly augmented CD86, CD54 and CD40 expression. These results revealed that the wide variety of responses to allergens in THP-1 cells may emulate allergen-induced maturation processes of DCs. It is suggested that THP-1 cells, which could develop several DC-like properties, are suitable for identifying sensitizing potential of chemicals. PMID- 17118623 TI - Assessing change with the Automated Neuropsychological Assessment Metrics (ANAM): issues and challenges. AB - Accurately documenting cognitive change is important, as neuropsychologists are routinely asked to determine cognitive change following disease progression or medical intervention. Computerized testing batteries, such as the Automated Neuropsychological Assessment Metrics (ANAM), are good tools for assessing change, because they allow for randomization of stimuli, creating near limitless alternate forms and reducing practice effects. The question remains, however, as to how best to determine reliable change in performance using ANAM. The current study compared the use of Reliable Change Index (RCI) and regression based methods (REG) calculated from 28 individuals with migraine. These methods then were applied to an independent sample of 25 individuals with migraine assessed with ANAM at baseline, headache, and following pharmacologic treatment. Traditional repeated measures analyses revealed declines in cognitive efficiency following migraine onset on two of four ANAM tasks and significant improvement on all ANAM tasks following treatment. Rates of deterioration and improvement did not significantly differ between RCI and REG methods, although were slightly different across the ANAM tasks used in this study. A combined ANAM score categorized the most individuals as demonstrating cognitive change, revealing that 60% of subjects declined in performance following headache and 84% improved following migraine treatment. PMID- 17118625 TI - Automated Neuropsychological Assessment Metrics sports medicine battery. AB - This paper describes the development and ongoing validation of the ANAM-sports medicine battery (ASMB) for use in concussion surveillance and management. A review of previous research utilizing the ASMB highlights current issues in concussion surveillance including: tracking cognitive recovery, effect of previous concussion history on acute concussion presentation, and clinical decision making using computerized measures. ASMB interpretation using reliable change indices or impairment indices is highlighted. Future development of the ASMB is discussed as it relates to interpretation of ASMB, development of appropriate norms, and defining adequate baseline assessment. This includes the definition of practice effects, the effects of maturation on test performance and definition of adequate baseline assessment that clearly defines a subject's normal cognitive performance level. ASMB is ready for cautious introduction into clinical practice for use by neuropsychologists with experience in both sports concussion and computerized testing. PMID- 17118626 TI - Broadening the definition of generalized anxiety disorder: effects on prevalence and associations with other disorders in the National Comorbidity Survey Replication. AB - Concerns have been raised that the DSM-IV requirements of 6-month duration, excessive worry, and three associated symptoms exclude a substantial number of people with clinically significant anxiety from a diagnosis of generalized anxiety disorder (GAD). We examined the implications of relaxing these three criteria for the estimated prevalence and predictive validity of GAD using nationally representative data from the US National Comorbidity Survey Replication. Relaxing all three criteria more than doubles the estimated prevalence of GAD. Broadly defined GAD significantly predicts the subsequent first onset of a wide range of temporally secondary disorders. The odds of secondary disorders are somewhat smaller for broadly defined than DSM-IV GAD, though few of these differences are statistically significant. Results suggest that subthreshold manifestations of GAD are significantly related to elevated risk of subsequent psychopathology. Further research is needed to determine whether broadening the current diagnostic criteria results in a more valid characterization of GAD. PMID- 17118627 TI - DNA microarray for detection of antibiotic resistance determinants in Bacillus anthracis and closely related Bacillus cereus. AB - We developed a multiplex PCR for amplification of ten genes involved in resistance to ciprofloxacin, doxycycline, rifampin, and vancomycin in Bacillus anthracis and closely related Bacillus cereus. Enzymatic labelling of PCR products followed by hybridization to oligonucleotide probes on a DNA microarray enabled simultaneous detection of resistance genes tetK, tetL, tetM, tetO, vanA, and vanB and resistance-mediating point mutations in genes gyrA, gyrB, parC, and rpoB. The presented assay allows detection of clinically relevant antibiotic resistance determinants within 4h and can be used as a time-saving tool supporting conventional culture-based diagnostics. PMID- 17118628 TI - Detection of regional blood perfusion changes in epileptic seizures with dynamic brain perfusion CT--a pilot study. AB - BACKGROUND AND PURPOSE: Perfusion CT (P-CT) is used for acute stroke management, not, however, for evaluating epilepsy. To test the hypothesis that P-CT may identify patients with increased regional cerebral blood flow during subtle status epilepticus (SSE), we compared P-CT in SSE to different postictal conditions. METHODS: Fifteen patients (mean age 47 years, range 21-74) underwent P-CT immediately after evaluation in our emergency room. Asymmetry indices between affected and unaffected hemispheres were calculated for regional cerebral blood volume (rCBV), regional cerebral blood flow (rCBF), and mean transit time (MTT). Regional perfusion changes were compared to EEG findings. RESULTS: Three patients in subtle status epilepticus (group 1) had increased regional perfusion with electro-clinical correlate. Six patients showed postictal slowing on EEG corresponding to an area of regional hypoperfusion (group 2). CT and EEG were normal in six patients with a first epileptic seizure (group 3). Cluster analysis of asymmetry indices separated SSE from the other two groups in all three parameters, while rCBF helped to distinguish between chronic focal epilepsies and single events. CONCLUSION: Preliminary results indicate that P-CT may help to identify patients with SSE during emergency workup. This technique provides important information to neurologists or emergency physicians in the difficult clinical differential diagnosis of altered mental status due to subtle status epilepticus. PMID- 17118629 TI - Crucial role of fibroblast integrins alpha2 and beta1 in maintaining the structural and mechanical properties of the skin. AB - BACKGROUND: In vivo functions of integrins in dermis have been investigated using several types of genetically integrin deficient mice. However, there are few studies to clarify actual in vivo functions of integrins in the dermis using normal type animals. OBJECTIVE: We investigated the actual in vivo functions of integrins in maintaining structural and mechanical properties in the normal skin by means of blocking interactions between fibroblasts and the extracellular matrix (ECM). METHODS: Intradermal injection of anti-integrin alpha2 or beta1 antibody into hairless rat skin was used to block the function of integrins. The dermal thickness was measured by an ultrasound scanner and the elastic properties of the skin was measured by Cutometer. RESULTS: Blocking integrin alpha2 or beta1 alone caused a moderate increase in dermal thickness. Blocking of integrins alpha1, alphaL or beta2 alone or blocking both integrins alpha1 and beta1 did not cause any change in the skin. However, blocking of both integrins alpha2 and beta1 caused a significant increase in dermal thickness accompanied by a marked loss of elastic properties. A clear change of the skin was observed within several minutes after injection, and continued for several hours. Treatment of human skin fibroblasts in collagen gel lattices with a mixture of anti-integrin alpha2 and beta1 antibodies in vitro caused marked and rapid morphological changes, but significant change was not observed with a treatment of alpha1, alpha2 or beta1 antibody alone. CONCLUSION: These results indicate that simultaneous functioning of integrins alpha2 and beta1 in fibroblasts play a crucial role in maintaining the structural and mechanical properties in the skin, which suggests that fibroblasts actively regulate collagen networks via these integrins. PMID- 17118630 TI - Protein adsorption on polymer-modified silica particle surface. AB - Silicon substrate surface and silica particle surface were modified with five kinds of polymers, poly(2-methoxyethyl methacrylate) (pMEMA), poly(2-hydroxyethyl methacrylate) (pHEMA), poly(acrylamide) (pAAm), poly(methyl methacrylate) (pMMA), and poly(styrene) (pSt), using a combined polymerization of surface-initiated polymerization that gives dense polymer chain layers and atom transfer radical polymerization (ATRP) that yields polymers with a narrow molecular weight distribution. Measurements of water contact angle and polymer chain amount on the modified silicon substrate surface and adsorption amounts of proteins (albumin and fibrinogen) on the modified silica particle surface revealed that the amount of polymer on the modified surface greatly affects the suppression of protein adsorption on the surface. PMID- 17118631 TI - Eudragit RS 100 microparticles containing 2-hydroxypropyl-beta-cyclodextrin and glutathione: physicochemical characterization, drug release and transport studies. AB - The aim of this study was to encapsulate glutathione (GSH) alone or in combination with hydroxypropyl-beta-cyclodextrin (HP-beta-CD) in Eudragit RS 100 microparticles (MPs), and to evaluate these novel delivery systems for oral administration of the considered tripeptide. The MPs were prepared by an O/O emulsion-solvent evaporation method according to a multilevel experimental design involving the volume of liquid paraffin, the HP-beta-CD amount, and the drug/polymer ratio as independent variables. The effects of these parameters on particle size, entrapment efficiency, and drug release were investigated. Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) analysis and differential scanning calorimetry (DSC) studies were performed to evaluate possible interactions between GSH and Eudragit RS 100 polymer and to characterize the physical state of drug within the MPs. The release profiles of GSH from MPs were examined in vitro at pH 1.2, 6.8. and 7.4 using the USP III (BioDis) dissolution apparatus. In general, a slow and zero-order release of GSH from MPs at pH 1.2 occurred, while at higher pH values considerable amounts of glutathione disulfide (i.e., GSSG) were observed. The enzymatic stability and the intestinal permeability of some GSH-containing MPs were assessed by using pepsin, alpha chymotrypsin, gamma-glutamyl-transpeptidase and everted frog intestinal sac methodology, respectively. The results suggest that GSH-loaded Eudragit RS 100 MPs containing HP-beta-CD represent a new sustained GSH delivery system useful for the oral administration of the examined tripeptide. PMID- 17118632 TI - [Pathologic childhood aerophagia]. AB - "Air swallowing" described as being part of functional gastrointestinal disorders in "Rome criteria" in 1999 is often misdiagnosed, particularly in non-mentally deficient children. AIMS: To recognize "air swallowing" child and to describe any progress according to the treatment. POPULATION AND METHODS: This retrospective study reports 13 cases of children without mental deficiency or neuromuscular disease. Clinical elements and precise histories are detailed and we have contacted consulting doctors or families for news. RESULTS: Ten boys and 3 girls, from 2,5 years to 10 years old, were referred for long lasting pain or abdominal distension. Numerous laboratory investigations were always normal. Diagnosis relied upon the observation of air swallowing and X-Rays views of gastric distension. Air swallowing was observed 7 times, 9 children had twitches and 3 language troubles. In 10 cases, X-rays showed gastric and colic distension. Three children have Chilaiditi syndrome. Favourable results followed in 12 cases after an average of 28 months of treatment. One case was lost for follow-up. Treatment was long, often disappointing and required the intervention of a psychiatrist, a paediatrician and (temporarily) a speech therapist. CONCLUSION: Pathological childhood aerophagia is often underdiagnosed and deserves to be better known by paediatricians, psychiatrists and surgeons. A late diagnosis leads to many negative results and causes anxiety. An early diagnosis should lead to a multidisciplinary care. PMID- 17118633 TI - [Breastfeeding and Cytomegalovirus infection in preterm infants. How to reduce the risks?]. AB - In France, screening for cytomegalovirus infection (CMV) during pregnancy is not recommended in routine. The transmission of CMV through breastmilk from mothers to preterm infants is frequent (15-20%). The frequency of neuro-sensorial handicap related to congenital CMV infection in very preterm infants is not well documented. We report the case of a female infant born at 30 weeks of gestation. At 15 days, she developed cholestatic jaundice. Urine cultures were positive for CMV. Diagnostic procedure showed no other cause for jaundice. At 40 days, the infant presented with hepato-splenomegaly, purpura and abnormal skin color related to a symptomatic, secondary CMV infection, probably transmitted through breastmilk. Ganciclovir was begun for 21 days. At 12 months, she presents with normal development. This observation raises questions about breastfeeding in very preterm infants. Unexplained prematurity could reflect recent infection or reactivation in the mother. Thus, because of the well-known risks of prematurity on one hand, and CMV infection on the other, we suggest that detection of CMV seropositive mothers should be considered before allowing breastfeeding. If the mother has serologic evidence of recent infection or reactivation, freezing breastmilk at -20 degrees C for 3 days may be an option in order to reduce virolactia, especially during early lactation. This may reduce the risk of postnatal vertical virus transmission with minimal logistical difficulties and without interrupting breastfeeding. PMID- 17118634 TI - [Community-acquired parapneumonic effusion in children: what's new?]. AB - Community-acquired pleural infection is a life-threatening complication of pneumonia in children. It seems to be more prevalent actually. This pathology is associated with an high morbidity and frequently requires prolonged hospitalization and invasives procedures. However, there is no consensus on its management in pediatrics, especially because of the lack of trials. To improve the quality of the future studies and to compare the series, a child-adapted classification is required. To date, in attempt of evidence, chest drainage or thoracocentesis-thoracoscopy are questionable. For treatment, high regimens of synergic and intravenously-delivered antibiotics seem to be the better choice. PMID- 17118635 TI - The long-term neurocompatibility of human fibrin sealant and equine collagen as biomatrices in experimental spinal cord injury. AB - INTRODUCTION: While fibrin sealant (FS) and equine collagen (EC) have been used as scaffold materials in experimental spinal cord injury (SCI), questions concerning neurocompatibility still remain. In this study, we assessed potential adverse effects, as well as functional and histological impact of FS and EC in subtotal hemisection of the thoracic spinal cord (SC) in rats. METHODS: 124 male rats were randomly assigned to four main groups (n=31): Sham (SH), Lesion only (L), fibrin sealant (GFS) and equine collagen group (GEC). SH animals received laminectomy only; all other animals underwent subtotal lateral hemisection at T9. Treatment consisted of application of FS or EC into the lesion gap in GFS and GEC, which was left empty in L. GFS, GEC, L and SH were each further divided into 4 subgroups: One subgroup, consisting of 10 rats was subjected to behavioural and reflex testing before surgery and followed up on days 1,7, 14, 21, 28 post op and then sacrificed. Haemalaun or cresyl violet (CV) was used to identify neutrophils in parasagittal cord sections which were obtained on day 1 (n=7). Sections stained for quantification of microglia/macrophages using ED-1 on day 3 (n=7), day 7 (n=7) and day 28 (n=7 out of 10). Additionally, neural filament (NF) staining was chosen to detect axonal regeneration and the length of ingrowth into FS and EC, Luxol blue for myelination, Von Willebrand factor for vascularisation, and glial fibrillary acidic protein (GFAP) staining for detection of astrocytes in glial scars on day 28. RESULTS: No adverse effects were observed in the treatment groups. Compared to L, GFS and GEC performed significantly better in the Basso, Beattie, Bresnahan (BBB) score and hopping responses. Proprioceptive placing was markedly improved in FS and EC compared to L. Axonal regrowth was found in GFS and GEC--the regrowth in the GFS was accompanied by myelination and vascularisation. Glial scarring occurred in all groups. Discussion Both biomatrices improved functional recovery compared to L and no adverse effects were perceived. PMID- 17118636 TI - Principles of intracellular viral recognition. AB - In mammalian cells, the products of microbial infection are recognized by pathogen recognition receptor (PRR) proteins. Virus recognition is mediated in part by PRRs that comprise a subset of Toll-like receptors or a family of RNA helicases, the latter of which contain caspase activation and recruitment domains, both of which induce interferons alpha and beta and antiviral immune defenses. Recent studies show that PRR engagement of specific pathogen-associated molecular patterns (PAMPs) within viral products, including viral proteins and nucleic acid, is facilitated by the discrete subcellular distribution of PRRs to sites that intersect with processes of virus entry and replication. PAMP structure and the subcellular context of PRR distribution form a basis of self versus nonself discrimination during the antiviral response. Understanding the virus/host interface of PRR function and PAMP recognition will advance therapeutic strategies for immune response regulation. PMID- 17118637 TI - Increased proportion of Fas positive CD8+ cells in peripheral blood of patients with COPD. AB - Chronic obstructive pulmonary disease (COPD) is characterised by chronic inflammation in pulmonary tissue and is also associated with systemic effects. The objective of this study was determination of lymphocyte subpopulation and the expression of Fas receptor on lymphocytes derived from peripheral blood of patients with stable COPD (n=18) and a control group: asymptomatic smokers (n=12) and non-smokers (n=12). Flow cytometry method with monoclonal antibodies was used for evaluation of lymphocyte subsets: CD4+ and CD8+ and the expression of Fas (CD95) on T lymphocytes. We found an elevated proportion of CD8+ cells in the blood of COPD patients. Proportion of Fas+ T lymphocytes was significantly higher in patients with COPD when compared with asymptomatic smokers and non-smokers (mean: 84.4% vs. 71.6% vs. 61.0% for Fas+/ CD4+ and 88.1% vs. 73.8% vs. 58.3% for Fas+/CD8+ lymphocytes). The proportion of Fas positive CD8+ cells significantly correlated with the degree of airway obstruction and hypoxemia. The significant correlations of Fas positive CD4+ and Fas positive CD8+ with smoking history expressed as pack years smoked were observed. Our observation of an elevated proportion of circulating lymphocytes bearing Fas receptor may play a role in induction of these cells' apoptosis and indicate the role of Fas/ FasL pathway in the changes in proportion of lymphocyte subpopulations in patients with COPD. PMID- 17118638 TI - Survival of patients on home mechanical ventilation: a nationwide prospective study. AB - Home mechanical ventilation (HMV) is increasingly used as a therapeutic option to patients with symptomatic chronic hypoventilation. There is, however, a paucity of solid data on factors that could affect prognosis in patients on home ventilation. In the present study, our aim was to study several factors in these patients with potential influence on survival. We examined 1526 adult patients from a nationwide HMV register to which data had been reported prospectively for 10 years. The patients constituted a broad diagnostic spectrum and the primary outcome in this study was death. We found by far the poorest survival rate in the ALS patients with only 5% alive after 5 years. Among the other patient groups the survival pattern was more uniform and the scoliosis, polio and Pickwick patients presented the best survival rate, after 5 years being around 75%. No factors were associated with a greater hazard for death in the ALS patients; in the non-ALS patients, however, negative predictors for survival were age, concomitant use of oxygen therapy, tracheostomy ventilation and start of ventilatory support in an acute clinical setting. Center size or county specific home ventilation treatment prevalence did not affect survival. In conclusion, in a large material of patients on HMV we found by far the poorest survival in the ALS patients. In the non-ALS patients a number of patient-related factors affected survival, while the size of the treating center or the regional treatment prevalence did not. PMID- 17118639 TI - Functional implications of BAL in the presence of restrictive or obstructive lung disease. AB - Most of the complications associated to bronchoscopy are related to changes of the respiratory function during or after its performance. Prevention of complications should be achieved by understanding the effects of bronchoscopic procedures and their relation to the pulmonary function deterioration. Previous studies regarding the functional impairment caused by bronchoalveolar lavage (BAL) were mostly limited by the presence of interferent factors such as sedative drugs. Furthermore, it is not clear whether or not patients with different ventilatory disturbances present the same functional response to bronchoscopy and BAL. The aim of this study was to determine the additional effects of BAL over the respiratory function deterioration related to bronchoscopy in patients with different respiratory function profiles (normal, restrictive and obstructive). Forty patients submitted to bronchoscopy without premedication were divided into four groups: group I-normal pulmonary function submitted to basic bronchoscopy; group II-bronchoscopy in combination with BAL, subdivided according to pulmonary function: group IIa (normal function), group IIb (restrictive ventilatory disturbances) and group IIc (obstructive ventilatory disturbances). Spirometry was made before and after the bronchoscopic procedure. Baseline hemoglobin saturation was compared to the lowest level during the procedure. Functional worsening caused by the procedure was observed with a decrease in forced vital capacity (FVC), forced expiratory volume in the first second (FEV(1)) and Hemoglobin saturation in all groups. Comparison between groups showed no significant difference regarding the changes in FVC (P=0.8324), FEV(1) (P=0.6952) and hemoglobin saturation (P=0.5044). We conclude that standardized BAL, like the one used in our study, does not result in an increased risk for ventilatory impairment compared to bronchoscopy itself, independently of the presence of previous respiratory disease. PMID- 17118640 TI - Recovery of PET from packaging plastics mixtures by wet shaking table. AB - Recycling requires the separation of materials appearing in a mass of wastes of heterogeneous composition and characteristics, into single, almost pure, component/material flows. The separation of materials (e.g., some types of plastics) with similar physical properties (e.g., specific gravity) is often accomplished by human sorting. This is the case of the separation of packaging plastics in municipal solid wastes (MSW). The low cost of virgin plastics and low value of recycled plastics necessitate the utilization of low cost techniques and processes in the recycling of packaging plastics. An experimental study was conducted to evaluate the feasibility of production of a PET product, cleaned from PVC and PS, using a wet shaking table. The wet shaking table is an environmentally friendly process, widely used to separate minerals, which has low capital and operational costs. Some operational variables of the equipment, as well as different feed characteristics, were considered. The results show that the separation of these plastics is feasible although, similarly to the mineral field, in somewhat complex flow sheets. PMID- 17118641 TI - Leaching characteristics of residual lateritic soils stabilised with fly ash and lime for geotechnical applications. AB - The effective use of residual lateritic soils as fill material for various construction activities is often limited by the difficulty in handling them. Attempt to improve their workability with fly ash and lime has shown promising results, although accompanied by risk of release of heavy metal leachates to the groundwater. In the present work, the leaching properties from residual lateritic soils from a part of Northeast India stabilised with fly ash and lime (CaO) was investigated with the help of single batch leaching test and column leaching test for different soil-fly ash-lime mixes. Test results show that the high pH induced by lime treatment of the mixes helps in keeping most of the metals within the stabilised soil matrix. Although the heavy metal concentrations in the leachates were generally within permissible limits, the release response for different metals was different suggesting implications for permeate solutions having metal pre-concentrations, such as those emanating from hazardous landfill sites. The observed characteristics provide insights towards the potential and realistic estimates of leaching of metals and its variation due to change in fly ash and lime content in the stabilised mix. Many of these constituents found in the stabilised soil had a first-flush phenomenon. But, as they occur only for short duration (about 5 pore volumes over 5 days) and at low concentration, dilution effect may eliminate them. The effect of continuous permeation on the flow parameters and the leaching pattern of the mixes have also been highlighted. PMID- 17118642 TI - Composting anaerobic and aerobic sewage sludges using two proportions of sawdust. AB - Sawdust has been proven to be a good bulking agent for sludge composting; however, studies on the most suitable ratio of sludge:sawdust for sludge composting and on the influence of the sludge nature (aerobic or anaerobic) on the composting reaction rate are scarce. In this study two different sewage sludges (aerobic, AS, and anaerobic, ANS) were composted with wood sawdust (WS) as bulking agent at two different ratios (1:1 and 1:3 sludge:sawdust, v:v). Aerobic sludge piles showed significantly higher microbial activity than those of anaerobic sludge, organic matter mineralization rates being higher in the AS mixtures. The lowest thermophilic temperatures during composting were registered when the anaerobic sludge was mixed with sawdust at 1:1 ratio, suggesting the presence of substances toxic to microorganisms. This mixture also showed the lowest decreases of ammonium during composting. All this matched with the inhibitory effect on the germination of Lepidium sativum seeds of this mixture at the first stages of composting, and with its low values of microbial basal respiration. However, the ANS+WS 1:3 compost developed in a suitable way; the higher proportion of bulking agent in this mixture appeared to have a diluting effect on these toxic compounds. Both the proportions assayed allowed composting to develop adequately in the case of the aerobic sludge mixture, yielding suitable composts for agricultural use. However, the ratio 1:1 seems more suitable because it is more economical than the 1:3 ratio and has a lower dilution effect on the nutritional components of the composts. In the case of the anaerobic sludge with its high electrical conductivity and ammonium content, and likely presence of other toxic and phytotoxic substances, the 1:3 ratio is to be recommended because of the dilution effect. PMID- 17118643 TI - DNA microarrays on silicon nanostructures: optimization of the multilayer stack for fluorescence detection. AB - To improve the sensitivity of fluorescence detection in DNA microarrays, the use of silicon nanostructures based on chemical vapor deposition (CVD) processes adopted for the growth of rough polycrystalline silicon was investigated. These substrates present advantages of two main properties which could lead to an enhancement of the fluorescence detection, i.e. (i) the increase of the available surface area in order to achieve a high loading capacity of biomolecules and (ii) the optimization of the stack of silicon nanostructures support. Indeed, the structures were elaborated on an initial thermal oxide layer and then covered with a silicon oxide layer, obtained by oxidation and allowing the functionalization for the subsequent grafting of DNA probes. Moreover, these oxide layers play a part in the fluorescence detection. The influence of the silicon oxide layer thickness above and below the silicon grains in close relation with the density of nanostructures on the emitted fluorescence was emphasized. This paper presents an experimental characterization of the fluorescence intensity and the optimization of the different layers that composed the substrate used for DNA microarrays. The performances of the microarrays were investigated by means of hybridization experiments using complementary fluorescent labeled-oligonucleotides targets. Our results indicate that an optimized substrate can be designed and that the use of oxidized silicon nanostructures for support of biochip could be a strategy for improving the sensitivity of fluorescence detection. PMID- 17118644 TI - Adsorption of 125I-labeled immunoglobulin G, its F(ab')2 and Fc fragments onto plasma-polymerized films. AB - Plasma-polymerized films were formed on flat glass plates using allylamine, acrylic acid, acrolein, and allylcyanide as monomers. Adsorption of (125)I labeled-proteins such as immunoglobulin G (IgG), its F(ab')(2) and Fc fragments, and human serum albumin (HSA) was measured on these plasma-polymerized (PP) films covering the glass plates and on commercially available polymer plates. The adsorption isotherm followed the Langmuir equation, from which the binding constant and amount of saturation binding were estimated. We found that, in general, a cationic surface had higher affinity for protein adsorption than an anionic surface. Among the surfaces examined, the PP-allylamine surface showed the highest binding capacity (264.2 nmol/m(2)) for F(ab')(2) fragment: it was remarkably high. Of the surfaces examined, the PP-acrylic acid surface showed the lowest binding capacity (12.8 nmol/m(2)) for F(ab')(2) fragment. The PP-acrylic acid surface also indicated the lowest protein binding capacity for IgG (16.5 nmol/m(2)), Fc-IgG (32.4 nmol/m(2)) and HSA (16.7 nmol/m(2)), respectively. These imply that the PP-acrylic acid film is useful to fabricate as a low protein adsorption material which expected to decrease cell adhesion. Results of our investigation indicate that the plasma-polymerization technique is promising for fabrication of a smart NanoBio-interface which can control the protein adsorption on a solid-phase substrate using a suitable monomer such as allylamine for the large adsorption and acrylic acid for the small adsorption. PMID- 17118645 TI - Electrochemical detection of 17beta-estradiol using DNA aptamer immobilized gold electrode chip. AB - An electrochemical detection method for chemical sensing has been developed using a DNA aptamer immobilized gold electrode chip. DNA aptamers specifically binding to 17beta-estradiol were selected by the SELEX (Systematic Evolution of Ligands by EXponential enrichment) process from a random ssDNA library, composed of approximately 7.2 x 10(14) DNA molecules. Gold electrode chips were employed to evaluate the electrochemical signals generated from interactions between the aptamers and the target molecules. The DNA aptamer immobilization on the gold electrode was based on the avidin-biotin interaction. The cyclic voltametry (CV) and square wave voltametry (SWV) values were measured to evaluate the chemical binding to aptamer. When 17beta-estradiol interacted with the DNA aptamer, the current decreased due to the interference of bound 17beta-estradiol with the electron flow produced by a redox reaction between ferrocyanide and ferricyanide. In the negative control experiments, the current decreased only mildly due to the presence of other chemicals. PMID- 17118646 TI - Methodology and evaluation of a highly sensitive algae toxicity test based on multiwell chlorophyll fluorescence imaging. AB - A new phytotoxicity bioassay based on chlorophyll fluorescence imaging of algae suspensions in multiwell plates is introduced. Phytotoxicity is quantified via inhibition of photosystem II quantum yield, Y(II), assessed with the saturation pulse method. The basics of this approach as well as the factors enhancing and limiting its performance are outlined. Compared to other established techniques the new system allows exceptionally rapid and accurate measurements of phytotoxicity using pulse-amplitude-modulation (PAM) fluorometry. While instrument related errors are negligibly small, optimal performance depends on appropriate choice of algae and illumination conditions. Illustrative examples for the response of Phaeodactylum tricornutum to diuron are presented. The standard deviation involved in the Y(II) determination of a single well amounts to the equivalent of 44 ng/L diuron. A decisive role is played by the light (measuring light, saturation pulses, actinic light) to which samples are exposed during the bioassay: (1) the inhibitor response is enhanced at high measuring light intensity. (2) Saturation pulses may be considered non-invasive only, if applied at low frequency and as long as physiologically healthy algae cultures are used. (3) Continuous actinic light may be problematic, as it induces complex physiological reactions that limit the performance of the approach; it is not required for assessment of diuron-type inhibitors at high measuring light intensity. PMID- 17118647 TI - The impact of the 'Clinical Trials' directive on the cost and conduct of non commercial cancer trials in the UK. AB - BACKGROUND: The UK Medicines for Human Use (Clinical Trials) Regulations 2004 implemented the European 'Clinical Trials' Directive (2001/20/EC) (EUCTD) into UK law and came into effect on 1(st) May 2004. In the period leading up to the implementation of the EUCTD in the UK there were serious concerns that it would have major cost implications for academic units running non-commercial clinical trials. METHODS: Directors and senior staff in 8 Clinical Trials Units (CTUs) were contacted and invited to participate in the study; arrangements were made for face-to-face interviews and the units were sent a questionnaire in advance of the meeting. The questionnaire was divided into six sections covering their involvement in non-commercial cancer clinical trials, and their perceptions of the EUCTD and its impact on all stages of trial development and conduct. Detailed cost data were also collected. FINDINGS: The findings from the questionnaire and interviews indicate that the EUCTD has resulted in a doubling of the cost of running non-commercial cancer clinical trials in the UK and a delay to the start of trials. The lack of central guidance, lack of clarity regarding the interpretation of the guidance notes, and increase in essential documentation and paperwork were causes of major concern for experienced staff who were anxious about whether they were interpreting the Directive correctly. Moreover, the CTUs were unable or unwilling to open trials in non-UK centres because of the different interpretation of the EUCTD by member states. INTERPRETATION: The EUCTD has both increased the cost and caused delay to non-commercial cancer clinical trials run by major public sector Clinical Trials Units in the UK. Staff have felt that they were working beyond capacity and were feeling demoralised in many CTUs. Finally, rather than harmonising and simplifying the regulatory environment, the Clinical Trials Directive has stopped many units from running trials in international centres. The UK has taken action to address some of the problems identified by this and other research, but problems remain. PMID- 17118648 TI - Histopathological risk factors for ipsilateral breast events after breast conserving treatment for ductal carcinoma in situ of the breast--results from the Swedish randomised trial. AB - AIM: The primary aims were to study risk factors for an ipsilateral breast event (IBE) after sector resection for ductal carcinoma in situ of the breast (DCIS) in a trial comparing adjuvant radiotherapy to no therapy and to assess predictive factors for response to radiotherapy. Secondary aims were to analyse reproducibility of the histopathological evaluation and to estimate correctness of diagnosis in the trial. SETTING: A randomised trial in Sweden (the SweDCIS trial), including 1046 women with a median of 5.2 years of follow-up in a population, offered routine mammographic screening. METHODS: A case-cohort design with a total of 161 cases of IBE (42 of those being members of the subcohort) and 284 sampled for the sub-cohort. Ninety five percent of the participants' slides could be retrieved and were re-evaluated by three experienced pathologists. RESULTS: Low nuclear grade (NG 1-2) and absence of necrosis halves the risk of IBE in both irradiated and non-irradiated patients. Lesion size, margins of excision and age at diagnosis did not modify these associations. The presence of necrosis modified the effect of radiotherapy: relative risk was 0.40 with necrosis present and 0.07 with necrosis absent (p-value for interaction 0.068). In all subsets of prognostic factors, radiotherapy conferred a substantial benefit. The risk factors for in situ and invasive IBE were similar. The agreement between pathologists was moderate (kappa=0.486). Correctness of diagnosis in the subcohort of SweDCIS was 84.8%. CONCLUSION: Although nuclear grade and necrosis carry prognostic information, we could not define a group with very low risk after sector resection alone. Radiotherapy has a protective effect in all substrata of risk factors studied. The interaction between the presence of necrosis and radiotherapy is a clinically and biologically relevant research area. PMID- 17118649 TI - Biosorption of cesium-137 and strontium-90 by mucilaginous seeds of Ocimum basilicum. AB - Mucilaginous seeds of Ocimum basilicum were used in uptake studies with cesium 137 and strontium-90. Results showed that uptake was dependent on the structural integrity of the mucilage fibrils. Water imbibed seeds showed higher adsorption of both 137Cs and 90Sr in comparison to seeds pretreated with NaOH, HCl and Na periodate solution. The uptake was pH dependent and while some divalent metal ions had no or little detrimental effect, the alkali metal ions Li+, Na+ and K+ decreased the uptake. The maximum adsorption capacity was 160 mg cesium g(-1) and 247 mg strontium g(-1) seed dry weight. PMID- 17118650 TI - Factors contributing to the poor bulk behavior of meat and bone meal and methods for improving these behaviors. AB - Meat and bone meal (MBM), a product of the rendering industry, is a potential feedstock for numerous bio-based applications. Design of processing equipment for MBM is difficult due to MBM's bulk behaviors; it flows less easily than many other granular materials, and it tends to foul the surfaces of processing equipment. This study examines the major factors contributing to MBM's poor bulk behavior, including moisture content, fat content, particle size distribution and temperature, and the relative importance of these factors. Potential methods for improving MBM's bulk properties, including use of an anti-caking agent, dehydration, fat extraction, milling and refrigeration are also studied. The effects of these factors were determined by a standard laboratory measurement, the Hausner ratio, as well as by the rate of surface-fouling and dust generation using a pilot-scale aspirator. In contrast to past studies with other granular materials, moisture content was shown to have an insignificant effect on MBM's bulk behavior. The results, however, show that MBM fat content is a major determinant of the bulk behavior of the MBM. Reduction of fat content resulted in major changes in MBM's bulk behavior, by all measures used. Less dramatic changes were achieved through refrigeration to solidify the fat and/or treatment with an anti-caking agent. PMID- 17118651 TI - Kinetics of batch ethanol fermentation of cheese-whey powder (CWP) solution as function of substrate and yeast concentrations. AB - A lactose utilizing yeast strain, Kluyveromyces marxianus DSMZ-7239 was used for ethanol formation from cheese-whey powder (CWP) solution in batch experiments. Effects of initial substrate (CWP) and yeast concentrations on the rate and extent of ethanol formation were investigated. The initial pH and oxidation reduction potential (ORP) was kept at 5 and -250 mV, respectively. The rate and extent of ethanol formation increased with increasing CWP concentration up to 156 g l(-1) (75 g l(-1) sugar) and then decreased for larger CWP concentrations due to substrate inhibition at high sugar concentrations. The ethanol yield coefficient was also maximum (0.54 g EtOH/g sugar) and equal to the theoretical yield at CWP concentration of 156 g l(-1). The growth yield coefficient was found to be Y(x/s)=1.2+/-0.1g biomass g sugar(-1). The rate of sugar utilization and ethanol formation also increased linearly with increasing initial biomass concentrations. A kinetic model describing the rate of sugar utilization and substrate inhibition as function of the initial substrate and the biomass concentrations was developed. The kinetic constants were determined using the experimental data. Model predictions of sugar utilization rates were in good agreement with the experimental data. The results indicated that the initial sugar concentration should be below 75 g l(-1) (CWP<156 g l(-1)) and the initial biomass should be above 850 mg l(-1) to obtain high rates and yields of ethanol formation and to avoid substrate inhibition. PMID- 17118652 TI - Highly constrained bicyclic VLA-4 antagonists. AB - VLA-4 is implicated in several inflammatory and autoimmune disease states. A series of cyclic beta-amino acids (beta-aa) was studied as VLA-4 antagonists. Binding affinity was highly dependent on the dihedral angle (phi) between the amino and the carboxyl termini of the beta-aa. Compound 5 m where the beta-aa is embedded in a bicycle possesses the most preferred phi (120 degrees). It is a potent and bioavailable VLA-4 antagonist (VCAM-Ig alpha4beta1 IC50 = 54 nM, rat po F = 49%). PMID- 17118653 TI - Intestinal permeability of antivirus constituents from the fruits of Eucalyptus globulus Labill. in Caco-2 Cell Model. AB - The uptake and transepithelial transport of the three main constituents macrocarpal A (M-A), macrocarpal B (M-B), and cypellocarpa C (Cy-C) from the fruits of Eucalyptus globulus Labill. were investigated. Monolayers of the human intestinal epithelial cancer cell line Caco-2 were incubated with M-A, M-B, and Cy-C to model its intestinal absorption and transport, respectively. The determination of compounds was performed by HPLC. The apparent permeability coefficients (P(app)) for M-A, M-B, and Cy-C in the apical-to-basolateral direction of a Caco-2 monolayer were (1.70+/-0.06)x10(-6), (1.99+/-0.10)x10(-6), and (6.08+/-0.41)x10(-6)cm/s, respectively. In the presence of iodoacetamide, the P(app) of Cy-C were both reducted in apical-to-basolateral and basolateral-to apical directions. M-A and M-B appear to accumulate in the epithelial cells. The intestinal absorption of M-A, M-B, and Cy-C was passive diffusion as the dominating process and Cy-C was partly ATP-dependent. PMID- 17118654 TI - Spirodiketopiperazine-based CCR5 antagonists: Lead optimization from biologically active metabolite. AB - Hydroxylated derivatives were designed and synthesized based on the information of oxidative metabolites. Compounds derived from beta-substituted (2R,3R)-2-amino 3-hydroxypropionic acid showed improved inhibitory activities against the binding of MIP-1alpha to human CCR5, compared with the non-hydroxylated derivatives and the other isomers. PMID- 17118655 TI - Alpha-methyltryptamine sulfonamide derivatives as novel glucocorticoid receptor ligands. AB - Alpha-methyltryptamine sulfonamides were identified as human glucocorticoid receptor (hGR) ligands in an ultra high throughput screening (UHTS) campaign. Described will be the hit-to-lead activities, including parallel and single point analog synthesis to map the scaffold. Ligands were identified that exhibited 30 nM binding to hGR. The SAR and selectivity of these compounds will be discussed. PMID- 17118656 TI - Enigmatic conflict of clinical and molecular diagnosis in Duchenne/Becker muscular dystrophy. PMID- 17118657 TI - Myosin storage (hyaline body) myopathy: a case report. AB - Myosin storage myopathy/hyaline body myopathy is a rare congenital myopathy, with less than 30 cases reported in the literature. It is characterised by the presence of subsarcolemmal hyaline bodies in type 1 muscle fibres and predominantly proximal muscle weakness. Recently, a single mutation (Arg1845Trp) in the slow/beta-cardiac myosin heavy chain gene (MYH7) was identified in four unrelated probands from Sweden and Belgium. The clinical severity and age of onset was variable, despite the same disease-causing mutation and similar histological findings. Here, we report the clinical and morphological findings of two brothers of English/Scottish background with the Arg1845Trp mutation in MYH7. This case report adds to the clinical description of this rare disorder and confirms that Arg1845Trp is a common mutation associated with this phenotype, at least in the White European population. PMID- 17118658 TI - Poloxamer 188 failed to prevent exercise-induced membrane breakdown in mdx skeletal muscle fibers. AB - We sought to determine the effectiveness of poloxamer 188 (P188) in protecting dystrophin-deficient, mdx skeletal muscle fiber membrane against exercise-induced breaches. mdx mice were treated with either P188 or placebo via intraperitoneal injections and run on a treadmill for 60-90 min. Membrane breakdown was quantified in cross-sections of rectus femoris muscle pretreated with Evans blue dye (in vivo). The mean % dye-penetrated muscle in the P188 and placebo groups was not significantly different in each of three trials. These results contrast with a recent report of P188 being highly effective in protecting the stretch- and dobutamine-stressed mdx heart muscle. The most likely explanations for the disparity are: (1) the exercise stress we used was beyond the protective range of P188, (2) P188 delivery and serum concentration were sub-optimal, or (3) the mdx skeletal myopathy and cardiomyopathy have fundamentally different responses to treatment. PMID- 17118659 TI - Describing physiotherapy interventions in an emergency department setting: an observational pilot study. AB - Physiotherapy interventions are provided to patients within Australian emergency departments in selected settings. By describing physiotherapy in the emergency department a contribution is made to understanding the shift in professional roles in emergency department settings. Using an intensive 5-day observational, single case design two key data sources were used: researcher observations and features of patient interventions provided by the emergency department (ED) physiotherapist outlined on a purposefully designed data collection sheet. Twenty patients (10 male, 10 female) were observed to have ED physiotherapy management during the data collection period. Physiotherapy interventions were targeted towards relieving pain, improving mobility, increasing joint range of motion and assisting with discharge planning of these patients. The interventions included educating patients about their conditions, providing gait aids, assisting with patient mobility and transfers, and liaising with medical, nursing and pharmacy staff for medication reviews and discharge planning requirements. Physiotherapy can assist with assessing and managing patients to contribute to reducing unnecessary hospital admissions from the ED. Interventions involve targeting features that would normally impede discharge of patients, such as reduced mobility, poor pain management and inadequate community or environmental supports, through interdisciplinary liaison with medical, nursing and allied health ED staff. PMID- 17118660 TI - Determinants of gait as applied to children with cerebral palsy. AB - In the present study, we quantified the isolated contributions of eight determinants of gait on the vertical center of mass (CoM) displacement of both typically developing children and children with cerebral palsy (CP). The role of these determinants, on vertical excursion, has never been examined for children or children with CP. We hypothesized that the relative contributions of the determinants to vertical CoM excursion of children with CP would be the same as the age-matched controls. We found that based on the similarities in the determinants effect on gait between the controls and adults reflect that children of this age walk with a mature gait. When applied to subjects with CP the determinant analysis found similar, but slightly exaggerated effects of those of the controls. All determinants that negatively affect CoM excursion were significantly worse in the children with CP, while those determinants that decreased excursion were varied. Heel rise, single support knee flexion, and pelvic obliquity had similar effects for on both groups. Pelvic rotation resulted in more excursion reduction in the controls, while leg inclination was more beneficial in reducing the CP groups excursion. The main cause for increased vertical excursion of the CoM in the children with CP was the increased knee flexion of both legs during double support. This excessive lowering of the CoM means that extra work is done to raise the CoM over the single support leg. The situation is aggravated by the fact that the CoM was lifted higher than typical because of the heel lifting during single support. Although these determinants allow quantification of the effects of gait kinematics and provide some useful information for gait they are limited in their ability to quantify the dynamics and kinetics of gait that are important for individuals with walking disabilities. PMID- 17118661 TI - Promising therapeutic agents for sepsis. AB - The incidence of sepsis is expected to increase at a rate of 1.5% per year. Advances in our understanding of the sepsis syndrome have enabled researchers to identify new therapeutic targets and design therapies for existing mediators of sepsis. Drotrecogin alfa (activated) was the first biological treatment for serious sepsis approved by the Food and Drug Administration in 2001. There have also been promising research results involving ethyl pyruvate, glycogen synthase kinase-3 inhibitors and 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors. Here, we review these four compounds and compound classes as examples of emerging pharmacological treatments of severe sepsis and describe the current status of sepsis research. PMID- 17118662 TI - Pyoverdine siderophores: from biogenesis to biosignificance. AB - Pyoverdines are a group of structurally related siderophores produced by fluorescent Pseudomonas species. Recent genomic and biochemical data have shed new light on the complex molecular steps of pyoverdine biogenesis and explained the chemical diversity of these compounds. In the opportunistic pathogen Pseudomonas aeruginosa, pyoverdine is necessary for infection in several different disease models. The occurrence of pyoverdine-defective strains in chronic infections of patients with cystic fibrosis and the extremely high sequence diversity of genes involved in pyoverdine synthesis and uptake indicate that pyoverdine production is subject to high evolutionary pressure. Pyoverdine dependent iron transport is also crucial for biofilm development, further expanding the importance of these siderophores in Pseudomonas biology. PMID- 17118663 TI - Transventricular hemispherotomy for surgical treatment of intractable epilepsy. AB - Surgical procedures for cerebral hemispherotomy may be broadly divided into those using a lateral and those using a vertical approach. However, careful study of surgical procedures using the lateral approach described in the literature shows differences in the approach to the ventricles. We discuss the application of transventricular hemispherotomy as a technique which provides relatively easy ventricular access for cerebral hemispherotomy. Transventricular hemispherotomy was successfully performed in a 36-year-old woman who was diagnosed with intractable epilepsy due to Sturge-Weber disease, and in a 25-year-old man who had developed intractable post-traumatic seizures after suffering cerebral contusion in a traffic accident as a child. These patients had no seizures or complications after surgery, and both patients have been weaned from antiepileptic drugs. The transventricular approach, as compared with other lateral approaches described in the literature, provides easy access to the ventricular cavity. Transventricular hemispherotomy proved to be a useful approach that allowed the following four common steps in cerebral hemispherotomy to be performed safely: (i) interruption of the internal capsule and corona radiata; (ii) resection of the medial temporal structures; (iii) transventricular corpus callosotomy; and (iv) disruption of the frontal horizontal fibers. PMID- 17118664 TI - N-Phenyl-N'-(2-chloroethyl)ureas (CEU) as potential antineoplastic agents. Part 2: role of omega-hydroxyl group in the covalent binding to beta-tubulin. AB - Tubulin is the target of many anticancer drugs, including N-phenyl-N'-(2 chloroethyl)urea (CEU). Unlike most anti-beta-tubulin agents, CEUs are protein monoalkylating agents binding through their N'-(2-chloroethyl)urea moiety to an amino acid nearby the colchicine-binding site on beta-tubulin isoform-2. Following the previously synthesized and attractive N-(3-omega hydroxyalkylphenyl)-N'-(2-chloroethyl)urea that exhibited growth inhibitory activity at the nanomolar level, we investigated the importance of lower alkyl and alkoxy groups to evaluate the effect of hydroxylated group and chain length on both cell growth inhibition and the mechanism of action of CEU. Here, we describe the preparation of two new series of CEU and show that the most potent CEU derivatives beside the omega-hydroxylated 1f were 2f and 3e, respectively. We have confirmed that the pentyl substituted CEUs 1f, 2f, and 3e are still covalently binding to beta-tubulin and still arrest cell division in G(2)/M phase. PMID- 17118665 TI - Paclitaxel binding to human serum albumin--automated docking studies. AB - A computational approach was used to study the interaction of the potent anticancer drug paclitaxel (Taxol) with human serum albumin. The primary and secondary binding sites were located at the interface of subdomains IIA and IIIA, and in the cleft between domains I and III of the protein, respectively. The C13 side chain and the baccatin core of paclitaxel were found to contribute approximately equally to the binding energy at the primary site, whereas the binding mode appears to be governed by the C13 side chain. PMID- 17118666 TI - Antennal morphology, structure and sensilla distribution in Microplitis pallidipes (Hymenoptera: Braconidae). AB - Microplitis pallidipes (Hymenoptera: Braconidae) is an important larval parasitoid of noctuid larval pests. In this study, we describe the morphology, ultrastructure and distribution of sensilla on the antennae of both male and female adults using scanning electron microscopy; complemented with transmission electron microscopy. Observations determined that the antennae of M. pallidipes were threadlike and the flagella were composed of 16 sub-segments. Totally, there were six types of sensilla found on the antennae. They were identified as Sensilla trichodea, Sensilla chaetica, Sensilla basiconica type I, S. basiconica type II, Sensilla coeloconicum, and elongated Sensilla placodea, respectively. The S. trichodea were the most abundant sensilla and distributed over the entire antennae, while S. basiconica I and II, S. coeloconicum, and elongated S. placodea were limited to the flagellum. S. chaetica were found on the scape and pedicle in very low numbers. The S. basiconica II and elongated S. placodea walls were full of pores as evident in transmission electron micrographs. Though the shape and structure of antennae between male and female adults were not basically different, the length and width of the antennae on male wasps were significantly greater than that of females. Also, the number, size and density of S. placodea of the male were significantly greater than those of females. The morphology and structure of these sensilla were compared with that found in other braconid wasps. Their possible functions are discussed in light of previously published literature. PMID- 17118667 TI - Determination of 210Po in environmental materials: a review of analytical methodology. AB - Polonium-210 ((210)Po) is analysed for a variety of purposes, including for radiological impact assessment, as a tracer of environmental processes, and as an indirect measure of its progenitor lead-210 ((210)Pb). Losses of polonium may occur at temperatures above 100 degrees C, depending on conditions, requiring particular care in sample preparation and treatment. In spite of this problem, the analysis of (210)Po is relatively straightforward, due to the ease of source preparation through spontaneous auto-deposition onto metal surfaces and the uncomplicated alpha spectrum. Although several optimisation studies have been carried out, published source preparation methods remain remarkably diverse. Some areas where further study would be useful are identified. PMID- 17118668 TI - Changes in the pool of soluble sugars induced by dehydration at the heterotrophic phase of growth of wheat seedlings. AB - Loss of dehydration tolerance coincides with a shift from heterotrophy to autotrophy during post-germination growth of spring wheat seedlings. This critical stage falls on the fifth day following imbibition. Till the sixth day of experiment light had no effect on dry weight of the seedlings but the survival of six day old seedlings was reduced by half upon dehydration. Germinating seeds in the presence of 5 mM glucose, fructose, mannose or sucrose did not promote seedling growth but either increase (glucose, fructose) or decreased (mannose, sucrose) the survival of dehydrated seedlings. Protection against dehydration by the former sugars was correlated, irrespective of the seedling age, with the decrease of sugar pool in seeds and increase in shoots (coleoptile and first leaf) and roots. The opposite changes were provoked by the sugars hampering seedling survival. Generally, survival of wheat seedlings was not correlated with the size of soluble sugar pool but its distribution and composition. Lower mobilisation of soluble sugars in seed, lower proportion of reduced sugars to sucrose and higher share of raffinose is characteristic for the tolerant four day old seedlings and those grown in the media containing glucose or fructose. The results presented indicate that higher proportion of reduced sugars to sucrose and lower share of raffinose in six day old seedlings seems to be associated with the loss of dehydration tolerance of these seedlings, despite heterotrophic character of growth. PMID- 17118669 TI - Video-assisted thoracic surgery lobectomy for lung cancer is associated with less immunochemokine disturbances than thoracotomy. AB - OBJECTIVE: Major surgery is immunosuppressive and could have an impact on postoperative tumor immunosurveillance and recurrence in cancer patients. Low circulating levels of insulin growth factor binding protein (IGFBP)-3 have been linked to advance prostate and the development of colonic cancers. This prospective study examined the early postoperative circulating levels of IGFBP-3, matrix metalloproteinase (MMP)-9, and tissue inhibitor of metalloproteinase (TIMP)-1 in early stage non-small cell lung cancer (NSCLC) patients undergoing major lung resection by VATS versus thoracotomy. METHODS: Forty-two consecutive patients with resectable primary NSCLC were assigned to VATS or thoracotomy approach over a 7-month-period. Blood samples were collected preoperatively and postoperatively on days (POD) 1 and 3 for enzyme linked immunosorbent assay determination of IGFBP-3, MMP-9 and TIMP-1 levels in the serum. RESULTS: There were no demographic differences between the two groups. VATS lung resection was associated with lower levels of MMP-9 and TIMP-1 on POD1 (median 628 vs 1311ng/ml, p=0.009; and 131 vs 211ng/ml, p=0.004, respectively) but higher levels of IGFBP-3 on POD3 (1366 vs 1144ng/ml, p=0.02), when compared with the thoracotomy approach. There was no perioperative mortality. CONCLUSIONS: VATS major lung resection for NSCLC is associated with higher circulating levels of IGFBP-3, and lower levels of MMP-9 and TIMP-1, compared to the thoracotomy approach. The clinical relevance of these postoperative changes on tumor biology following lung resection for cancer warrants further investigation. PMID- 17118670 TI - Intrapulmonary lymph node metastasis and renal cell carcinoma. AB - Intrapulmonary lymph node metastasis occuring after nephrectomy for renal cell carcinoma is a particular recurrence modality. Authors report two male patients presenting with this recurrence 2 and 4 years after treatment of the primary and who underwent surgery. Surgery confirmed the diagnosis and demonstrated station 7 minimal metastases and positive pleural lavage cytology in both patients. The first patient survived 2 years and the second was alive disease free at 33 months follow-up. Such metastases probably originate from the thoracic duct. Resection confirms the diagnosis and may be part of the treatment. PMID- 17118671 TI - Direct comparison of Au(3)(+) and C(60)(+) cluster projectiles in SIMS molecular depth profiling. AB - The sputtering properties of two representative cluster ion beams in secondary ion mass spectrometry (SIMS), C(60)(+) and Au(3)(+), have been directly compared. Organic thin films consisting of trehalose and dipalmitoylphosphatidylcholine (DPPC) are employed as prototypical targets. The strategy is to make direct comparison of the response of a molecular solid to each type of the bombarding cluster by overlapping the two ion beams onto the same area of the sample surface. The ion beams alternately erode the sample while keeping the same projectile for spectral acquisition. The results from these experiments are important to further optimize the use of cluster projectiles for SIMS molecular depth profiling experiments. For example, Au(3)(+) bombardment is found to induce more chemical damage as well as Au implantation when compared with C(60)(+). Moreover, C(60)(+) is found to be able to remove the damage and the implanted Au effectively. Discussions are also presented on strategies of enhancing sensitivity for imaging applications with cluster SIMS. PMID- 17118672 TI - Use of axenic animals in studying the adaptation of mammals to their commensal intestinal microbiota. AB - Vertebrates are essentially born germ-free but normally acquire a complex intestinal microbiota soon after birth. Most of these organisms are non pathogenic to immunocompetent hosts; in fact, many are beneficial, supplying vitamins for host nutrition and filling the available microbiological niche to limit access and consequent pathology when pathogens are encountered. Thus, mammalian health depends on mutualism between host and flora. This is evident in inflammatory conditions such as inflammatory bowel disease, where aberrant responses to microbiota can result in host pathology. Studies with axenic (germ free) or deliberately colonised animals have revealed that commensal organisms are required for the development of a fully functional immune system and affect many physiological processes within the host. Here, we describe the technical requirements for raising and maintaining axenic and gnotobiotic animals, and highlight the extreme diversity of changes within and beyond the immune system that occur when a germ-free animal is colonized with commensal bacteria. PMID- 17118673 TI - High density fermentation and activity of a recombinant lumbrokinase (PI239) from Pichia pastoris. AB - A system for the expression of recombinant lumbrokinase (rPI239) was developed in the yeast Pichia pastoris. A total supernatant protein content of 0.174 g/L of high density fermentation broth was obtained. The rPI239 exhibited in vitro fibrinolytic activity. The in vivo activity of rPI239 was measured by prothrombin time, kaolin part thrombin time, thrombin time, and fibrinolytic activity. This work presents the high-density fermentation of rPI239 from P. pastoris and shows that the recombinant protein has similar fibrinolytic activity both in vivo and in vitro. PMID- 17118674 TI - Protection of Procambarus clarkii against white spot syndrome virus using recombinant oral vaccine expressed in Pichia pastoris. AB - The potential for oral vaccination of crayfish against white spot syndrome virus was investigated. The envelope proteins VP19 and VP28 were expressed in yeast (Pichia pastoris). The expressed proteins were used as oral vaccines in different forms viz., in whole culture form, whole culture sonicated form, whole culture centrifuged supernatant form, and cell residue form. The recombinant proteins were mixed with food pellets and fed to crayfish for 25 days. The vaccinated groups were divided into two even groups and challenged on the 3rd and 21st day of post vaccination. Among different vaccine groups the relative percent survival (RPS) values of sonicated form and supernatant form vaccines were found the best and met the criterion (>RPS 60%) of effective vaccine even after 21st day of post vaccination. Development of vaccine by using recombinant proteins VP19 and VP28 in yeast as expression vector was feasible with significant effects. PMID- 17118675 TI - Phylogenetic relationships of egg parasitoids (Hymenoptera: Eulophidae) and correlated life history characteristics of their Neotropical Cassidinae hosts (Coleoptera, Chrysomelidae). AB - Egg parasitoids in the family Eulophidae (Hymenoptera) are an important part of the community of insects attacking neotropical leaf beetles in the subfamily Cassidinae. We present a phylogeny of 24 species of oophagous Eulophidae, using the 28S rDNA, the ITS2 rDNA and the cytochrome b genes, applying the NJ, MP, ML and Bayesian tree reconstruction methods on each data set. We ask whether the phylogenetic relationships of the parasitoids are linked with the life history characteristics of their beetle hosts. We show that cladogenesis in the oophagous Eulophidae does correlate with ovipositional behaviour and, to a lesser extent, diet and tribal affinities of their hosts. Additionally using two methods of simultaneous analysis of several gene sets: the Total Evidence method, and the construction of a "supertree" by Matrix Representation Parsimony (MRP), we substantiate the same major phylogenetic relationships within the Eulophidae. PMID- 17118676 TI - Late onset of axillary artery thrombosis after a nondisplaced humeral neck fracture: a case report. PMID- 17118677 TI - The pharmacokinetics of vigabatrin in rat blood and cerebrospinal fluid. AB - PURPOSE: Data on the blood pharmacokinetics of vigabatrin, an antiepileptic drug with a unique and novel mechanism of action, in the rat are sparse. Additionally, little is known of the kinetics of vigabatrin in the central cerebrospinal fluid (CSF) compartment. We therefore investigated the rate of penetration into and the inter-relationship between serum and CSF compartments following systemic administration of vigabatrin in the rat. METHODS: Sprague-Dawley rats were implanted with a jugular vein catheter and a cisterna magna catheter for blood and CSF sampling, respectively. Vigabatrin was administered by intraperitonial injection at three different doses (250, 500 and 1000mg/kg) and blood and CSF collected at timed intervals up to 8h. Vigabatrin concentrations in sera and CSF were determined by high performance liquid chromatography. RESULTS: Vigabatrin concentrations in blood and CSF rose linearly and dose-dependently and the time to maximum concentration (Tmax) was 0.4 and 1.0h, respectively. Vigabatrin is not protein bound in serum and its elimination from serum (mean t1/2 values, 1.1-1.4 h) is rapid and dose-independent. The efflux of vigabatrin from CSF was significantly slower than that seen for serum (mean t1/2 values, 2.2-3.3h). CONCLUSIONS: The kinetics of vigabatrin are linear with rapid entry into CSF. However, although vigabatrin CSF kinetics parallel that seen in serum, CSF vigabatrin concentrations represent only 2% of concentrations seen in serum and do not reflect free drug concentrations in serum. PMID- 17118678 TI - Microinjections of the dopamine D2 receptor antagonist sulpiride into the medial prefrontal cortex attenuate glucocorticoid-induced impairment of long-term memory retrieval in rats. AB - We recently reported that blockade of dopamine (DA) D2 receptors attenuated deficits in long-term memory retrieval induced by a systemic injection of corticosterone, but the anatomical sites of such interaction were not known. In this study, we investigated whether the DA D2 receptors located in the medial prefrontal cortex (mPFC) may play a role in the impairing effects of glucocorticoids on the memory retrieval process. Young adult male rats were trained in a one trial inhibitory avoidance task (0.5 mA, 3s footshock). On the retention test given 48 h after training, the latency to re-enter the dark compartment and the time spent in light compartment of the apparatus were recorded. Systemically administered corticosterone (1mg/kg) given to rats 30 min before retention testing impaired their memory retrieval. Bilateral microinjections of the DA D2 receptor antagonist sulpiride (10 or 100 ng/0.5 microl per side) into the mPFC 30 min before corticosterone administration attenuated the glucocorticoid-induced impairment of memory retrieval. Furthermore, applied doses of sulpiride alone were ineffective in modulating memory retrieval. These findings indicate that D2 receptors located in the mPFC play an important role in mediating the impairing effects of glucocorticoids on memory retrieval. PMID- 17118679 TI - The protein kinase C orthologue PkcA plays a role in cell wall integrity and polarized growth in Aspergillus nidulans. AB - The calC2 mutation in Aspergillus nidulans causes hypersensitivity to Calcofluor White, along with other drug sensitivities that indicate a defect in cell wall integrity. We have cloned CalC by complementation, isolating the A. nidulans orthologue of protein kinase C (PkcA). The pkcA allele of the calC2 strain contains a mutation predicted to introduce a charged arginine residue in place of neutral glycine at a conserved site located immediately beside the C1B regulatory domain. Both PkcA and calC2 map to the same region of chromosome VIII. A PkcA::GFP chimera localizes to hyphal apices and growing septa, as well as to the conidiogenous apices of phialides, indicating a role for PkcA in polarized cell wall growth. These observations support the hypothesis that the role of PkcA in A. nidulans, is comparable to that played by Pkc1p in the Saccharomyces cerevisiae cell wall integrity pathway. PMID- 17118680 TI - Acute muscle inflammation enhances the monosynaptic reflexes and c-fos expression in the feline spinal cord. AB - The aim of this research was to study the changes of the motor reflex activity (monosynaptic reflex (MSR) of the flexor and extensor muscles) and Fos immunoreactivity in lumbo-sacral spinal cord after acute induced myositis of m. gastrocnemius-soleus (GS). The experiments were carried out on ischaemic decerebrated, spinalized in C1 cats. After infiltration of the GS muscle with carrageenan (2%) MSRs of flexors and extensors showed a significant increase in amplitude +127+/-24.5% and +155+/-28.5%, respectively, p<0.05. The exposed effect was initiated within 30 min and achieved a maximum 2.8h after the intramuscular injections of carrageenan. After analysis of dynamics of the MSRs, animals were perfused and c-fos expression in the spinal segments L6-S1 was evaluated. In comparison to sham-operated animals, the number of Fos-immunoreactive (Fos-ir) cells was noticeably increased in the lumbar cord of cats with carrageenan induced myositis. The labeled cells were concentrated in the ipsilateral laminae I/II, neck of the dorsal horn (V/VI) and intermediate zone (VII), however, clear predominance of their concentration was found in the deep laminae. The effect of muscle inflammation was also expressed as a significant decline in the number of NADPH-d-reactive cells (p<0.05) in ipsilateral laminae I/II of L6/L7. The results show that the input from acutely inflamed muscles may induce an increase of the reflex responsiveness of flexors and extensors which is not mediated via the gamma-spindle-loop and which coincides with a significant increase in c-fos expression in the deep laminae of the lumbar spinal cord. PMID- 17118681 TI - Self-reported pain sensitivity: lack of correlation with pain threshold and tolerance. AB - Many recent studies and several reviews have highlighted the potential clinical applications of experimental pain testing (e.g., for predicting post-surgical pain, treatment responsiveness, etc.). However, the implementation of quantitative sensory testing of pain sensitivity on a broad scale is limited by requirements of time, equipment, and expertise, and their associated costs. One reasonable question is whether one can obtain, via self-report, a valid index of an individual's pain sensitivity and pain tolerance. We analyzed data from a large number of subjects (n=505) who had undergone standardized thermal pain testing, and found that while higher self-reported pain sensitivity was associated with higher scores on a measure of anxiety, no relationship was observed between subjects' self-report of pain sensitivity and subjects' actual pain threshold or tolerance. These findings suggest that circumventing psychophysical pain testing by assessing individuals' self-reported pain sensitivity is unlikely to be a useful strategy. PMID- 17118682 TI - Long-term reliable change of pain scores in individual myogenous TMD patients. AB - A within-patient change in pain score after treatment is statistically 'reliable' when it exceeds the smallest detectable difference (SDD). The aims of the present study were to: (i) determine SDDs for VAS-scores of pain intensity, for sufficiently long test-retest intervals to include most biological fluctuations, (ii) examine whether SDD is invariant to baseline score, and (iii) discuss the value of reliable change (RC) for detecting clinically important difference (CID) or as a possible indicator of successful treatment. SDDs were determined using duplicate data from 118 patients with myogenous Temporomandibular disorders: (1) VAS-scores of pain intensity from the masticatory system in a pre-treatment diary, and (2) VAS-scores of pain intensity from the hand (cold-pressor test). RC was determined in VAS-scores from a pre- and post-treatment questionnaire. The long-term SDD was 49mm. A regression analysis on duplicate VAS-scores showed that SDD was largely invariant to the baseline level. Because RC (change>SDD) exceeded CID, it might serve as an indicator of successful treatment. However, only 17% of the patients showed RC after treatment, mainly because the baseline was smaller than SDD in 67% of the patients thus making detection of any treatment effect impossible. For patients with possible detection (33%), the frequency of RC was 51%. If the detection threshold would be avoided by provoking pain in patients with a low baseline, a long-term RC in VAS-scores might occur in about half of all myogenous TMD patients and might then serve as an indicator of cases of treatment success. PMID- 17118683 TI - Fast nosologic imaging of the brain. AB - Magnetic resonance spectroscopic imaging (MRSI) provides information about the spatial metabolic heterogeneity of an organ in the human body. In this way, MRSI can be used to detect tissue regions with abnormal metabolism, e.g. tumor tissue. The main drawback of MRSI in clinical practice is that the analysis of the data requires a lot of expertise from the radiologists. In this article, we present an automatic method that assigns each voxel of a spectroscopic image of the brain to a histopathological class. The method is based on Canonical Correlation Analysis (CCA), which has recently been shown to be a robust technique for tissue typing. In CCA, the spectral as well as the spatial information about the voxel is used to assign it to a class. This has advantages over other methods that only use spectral information since histopathological classes are normally covering neighbouring voxels. In this paper, a new CCA-based method is introduced in which MRSI and MR imaging information is integrated. The performance of tissue typing is compared for CCA applied to the whole MR spectra and to sets of features obtained from the spectra. Tests on simulated and in vivo MRSI data show that the new method is very accurate in terms of classification and segmentation. The results also show the advantage of combining spectroscopic and imaging data. PMID- 17118684 TI - Comparable spirometric efficacy of tiotropium compared with salmeterol plus fluticasone in patients with COPD: a pilot study. AB - BACKGROUND: International guidelines recommend the long-acting anticholinergic, tiotropium, or long-acting beta 2-agonists as maintenance therapy in patients with moderate-to-very severe chronic obstructive pulmonary disease (COPD). The efficacy of long-acting beta(2)-agonists combined with inhaled corticosteroids (ICS) in the treatment of COPD has also been confirmed for severe and very severe COPD, but data comparing tiotropium with the combination of a long-acting beta 2 agonist and an ICS are lacking. METHODS: This 6-week multicentre, randomised, double-blind, triple-dummy pilot study compared the bronchodilator effects of tiotropium 18 microg once daily (n=56) vs. the combination of salmeterol 50 microg plus fluticasone 250 microg twice daily (n=51) in patients with moderate to-very severe COPD. Serial spirometry was performed over 12h after 6 weeks of treatment. The primary endpoint was forced expiratory volume in 1s (FEV1) area under the curve from 0 to 12h (AUC0-12h) on Day 43. RESULTS: Randomization failed to provide treatment groups with comparable baseline characteristics for smoking history, current smokers, duration of COPD, FEV1, forced vital capacity (FVC) and reversibility. Mean+/-SD FEV1 was 1.31+/-0.47 l in the tiotropium group vs. 1.46+/-0.53 l in the salmeterol plus fluticasone group. Fewer patients in the tiotropium showed a 12% and 200 ml acute increase to short-acting bronchodilators at baseline. However, treatment with tiotropium alone resulted in comparable bronchodilation compared with salmeterol plus fluticasone, as measured by all the spirometric parameters at the end of the 6-week study period. FEV1 AUC0-12h was 1.55+/-0.03 l in the tiotropium group vs. 1.57+/-0.04 l in the salmeterol plus fluticasone groups (p=0.63). Trough (predose) FEV1 was 1.54+/-0.03 l in the tiotropium group vs. 1.46+/-0.03 l in the combination group (p=0.07), and peak FEV(1) was 1.68+/-0.04 l vs. 1.66+/-0.04 l, respectively, (p=0.77). FVC AUC0-12h, trough and peak were also comparable between groups at study end (p>0.05, for all). Further, rescue salbutamol use was similar in the tiotropium and combination groups and both treatment regimens were well tolerated. CONCLUSIONS: Six weeks of treatment with tiotropium resulted in comparable bronchodilation compared with salmeterol plus fluticasone in patients with moderate-to-very severe COPD, despite tiotropium patients having lower lung function and fewer patients considered reversible at baseline. The results of this pilot study will aid planning for further large-scale comparative studies. PMID- 17118685 TI - Cough throughout life: children, adults and the senile. AB - We review cough from premature birth, mature neonatal life, in childhood and adult life, and in old age. There is a regrettable lack of definitive studies, but many clues in the literature. The cough reflex seems weak in premature infants, but develops with maturity. It is pronounced in childhood, but there seem to be no studies comparing its strength then with that in adulthood. In old age the cough may weaken, as indicated by the prevalence of aspiration pneumonia. These changes are presumably related to the development and degeneration of the afferent and central nervous pathways for cough, which may be reflected in the changes in laryngeal muscle function with age. There is much evidence that age influences the development of the respiratory system in general, and of the immune system which would affect the degree, frequency and clinical issues of cough. Other factors that limit our understanding of the changes in cough with age include the reporting of cough by parents in infants and carers in old age and the use of different diagnostic criteria throughout life. Age-related variation in cough sensitivity seems to be well established, but its quantitation and mechanisms require much further research. PMID- 17118686 TI - Comparable long-term safety and efficacy of a novel budesonide/formoterol pressurized metered-dose inhaler versus budesonide/formoterol Turbuhaler in adolescents and adults with asthma. AB - Budesonide/formoterol in one inhaler is an established therapy for asthma and chronic obstructive pulmonary disease. The long-term safety and efficacy profile of a novel hydrofluoroalkane (HFA) pressurized metered-dose inhaler (pMDI) formulation of budesonide/formoterol was compared with that of budesonide/formoterol in a dry powder inhaler (DPI; Turbuhaler). This multinational, 52-week, randomized, open, parallel-group study included patients aged > or = 12 years with asthma who had a forced expiratory volume in 1s (FEV1)> or = 50% of predicted normal; all patients used inhaled corticosteroids (400-1200 microg/day) and needed additional short-acting beta 2-agonist therapy. Patients were randomized to receive budesonide/formoterol pMDI or DPI 160/4.5 microg, two inhalations twice daily. Safety endpoints included assessment of adverse events and laboratory parameters. Efficacy endpoints included change from baseline in FEV1 and time to first severe asthma exacerbation. Overall, 673 patients (446pMDI, 227DPI) were included. There were no clinically significant differences between treatment groups in the nature, incidence or severity of adverse events or laboratory parameters. The number of patients experiencing adverse events was comparable in the pMDI (332/446 [74%]) and DPI (175/227 [77%]) groups; the most commonly reported adverse event was upper respiratory tract infection. The proportion of patients discontinuing as a result of adverse events was low in both groups (pMDI 12/446 [3%], DPI 2/227 [1%]). Lung function was improved to a similar extent in both groups and there was no detectable difference in time to first severe asthma exacerbation. The novel HFA pMDI formulation of budesonide/formoterol is an equally well tolerated and effective treatment for adults and adolescents with asthma as the budesonide/formoterol DPI. PMID- 17118687 TI - The effects of recombinant human growth hormone (rhGH) supplementation on adipokines and C-reactive protein in obese subjects. AB - OBJECTIVE: Obese subjects have functional growth hormone deficiency (GHD). Recombinant human GH (rhGH) treatment of pituitary GHD improves serum levels of leptin, adiponectin and C-reactive protein (CRP). This study was undertaken to determine whether these rhGH-induced changes occur in obese subjects during rhGH supplementation. DESIGN: Randomized double-blind placebo-controlled trial of low dose rhGH (200 microg/day for the first month, then 400 microg/day for men and 600 microg/day for women thereafter) or placebo supplementation as an adjuvant to a standard weight loss program. SUBJECTS: Forty healthy obese subjects, 28 premenopausal menstruating women (35+/-7 SD years) and 12 men (37+/-6 years). MEASUREMENTS: Body weight, BMI, body composition (assessed by dual energy X-ray absorptiometry [DEXA]), and serum levels of glucose, insulin, IGF-I, IGFBP-3, insulin resistance index (homeostasis modal assessment [HOMA]), leptin, CRP and adiponectin were performed at baseline and at 6 months. RESULTS: For similar entry BMI values, women when compared with men had higher percent body fat (BF) (43.5+/-4.6% vs. 29.8+/-4.0%, p<0.001), higher leptin levels (16.9+/-8.4 microg/L vs. 4.2+/-3.0 microg/L, p<0.001), and higher CRP levels (13.8+/-16.8 mg/L vs. 2.4+/-3.2mg/L, p=0.04). Serum levels of leptin and CRP, but not adiponectin, correlated significantly with BF in both sexes. Recombinant human GH treatment increased levels of IGF-I Z-Score between baseline and 6 months (from -0.7+/-0.9 SD to 0.1+/-1.1 SD, p=0.01) and modestly decreased BF (from 38.4+/-7.8% to 35.6+/ 7.5%, p=0.046). Despite increased IGF-I, there were no differences between rhGH and placebo with regard to changes in leptin, CRP, or adiponectin. CONCLUSION: It is concluded that in obesity, although rhGH treatment significantly increases IGF I and modestly reduces body fat, the lack of significant changes in serum leptin, adiponectin or CRP levels suggests that rhGH treatment does not have a significant effect on these serum markers of adiposity. PMID- 17118688 TI - Respective contributions of vertebroplasty and kyphoplasty to the management of osteoporotic vertebral fractures. AB - Among individuals aged 50-80 years, 5-20% have one or more vertebral crush fractures. One-third of these patients require treatment for acute or chronic pain. Vertebroplasty and kyphoplasty were introduced over the last two decades as treatments for incapacitating pain from osteoporotic vertebral fractures. Both techniques proved effective and safe in numerous retrospective and prospective studies. They now deserve to be incorporated into the standard treatment strategy for painful and incapacitating vertebral fractures. Kyphoplasty seeks not only to stabilize the vertebra, but also to correct the kyphosis induced by the vertebral body collapse. However, the correction is often limited (less than 15 degrees ) and has not been shown to increase the benefits in terms of pain relief or quality-of-life improvement, compared to vertebroplasty. Kyphoplasty is more costly than vertebroplasty, which is therefore emerging as the treatment of choice. However, a randomized double-blind trial comparing vertebroplasty and kyphoplasty is needed. Furthermore, a randomized comparison of vertebroplasty or kyphoplasty versus noninterventional treatment is needed in patients admitted for pain immediately after a vertebral crush fracture. PMID- 17118689 TI - Comments on the original article by Christine Albert et al.: "septic knee arthritis after intra-articular hyaluronate injection. Two case reports". PMID- 17118690 TI - [Is tubal and uterine surgery deleterious to ovarian reserve?]. AB - Changes in menstrual pattern after tubal sterilisation have been reported for more than 50 years. Hence all tubal surgeries have been suspected of altering the ovarian reserve, by damage to the ovarian blood vessels. Recent studies showed that tubal surgery has no significant adverse effect on doppler flow indice and hormonal markers. Hysterectomy and uterine artery embolization seem to decrease ovarian reserve in perimenopausal women. Uterine artery embolization does not seem to have adverse effects on normally functioning ovaries. PMID- 17118691 TI - Design and construction of small sized pencil probe to measure bio-impedance. AB - Currently, bio-impedance measurements are performed with relatively large probes which are not suitable for all in vivo studies. These are typically designed and constructed for different uses, such as for cervical and oesophagus tissues and are too large for many investigations, including those involving the bladder. Therefore, it was decided to design and construct a small sized pencil probe, using a microscope to solder very small wires to a tiny tip (about 2mm in diameter). In addition, different approaches were used to construct, treat, and perform the safety tests and calibration procedure on the probe before taking impedance measurements of the urinary bladder. PMID- 17118692 TI - Deletion of JAM-A causes morphological defects in the corneal epithelium. AB - Junctional adhesion molecule-A (JAM-A, JAM-1, F11R) is an Ig domain containing transmembrane protein that has been proposed to function in diverse processes including platelet activation and adhesion, leukocyte transmigration, angiogenesis, epithelial cell shape and endothelial cell migration although its function in vivo is less well established. In the mouse eye, JAM-A protein expression is first detected at 12.5 dpc in the blood vessels of the tunica vasculosa, while it is first detected in both the corneal epithelium and lens between 13.5 and 14.5 dpc. In the corneal epithelium, JAM-A levels remain appreciable throughout life, while JAM-A immunostaining becomes stronger in the lens as the animals age. Both the cornea and lens of mice lacking an intact JAM-A gene are transparent until at least a year of age, although the cells of the JAM A null corneal epithelium are irregularly shaped. In wild-type mice, JAM-A protein is found at the leading edge of repairing corneal epithelial wounds, however, corneal epithelial wound repair was qualitatively normal in JAM-A null animals. In summary, JAM-A is expressed in the corneal epithelium where it appears to regulate cell shape. PMID- 17118693 TI - Peroxisome proliferator-activated receptor gamma (PPARgamma) regulates trefoil factor family 2 (TFF2) expression in gastric epithelial cells. AB - Although trefoil factor family 2 (TFF2) plays a critical role in the defense and repair of gastric mucosa, the regulatory mechanism of TFF2 expression is not fully understood. In this study, we investigated the regulation of TFF2 expression by peroxisome proliferator-activated receptor gamma (PPARgamma) in gastric epithelial cells. MKN45 gastric cells were used. TFF2 mRNA expression was analyzed by real-time quantitative RT-PCR. The promoter sequence of the human TFF2 gene was cloned into pGL3-basic vector for reporter gene assays. Ciglitazone was mainly used as a specific PPARgamma ligand. MKN45 cells expressed functional PPARgamma proteins. Endogenous TFF2 mRNA expression and TFF2 reporter gene transcription was significantly up-regulated by ciglitazone in a dose-dependent manner. Reporter gene assays showed that two distinct cis-elements are involved in the response to PAPRgamma activation. Within one of these element (nucleotides -558 to -507), we identified a functional peroxisome proliferator responsive element (PPRE) at -522 (5'-GGGACAAAGGGCA-3'). Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) assay confirmed the binding of PPARgamma to this sequence. Another element (nucleotides -407 to -358) appeared to be a composite enhancer element indirectly regulated by PPARgamma and a combination of these two cis-elements was required for the full response of the human TFF2 gene expression to PPARgamma. These data demonstrate that human TFF2 gene is a direct target of PPARgamma in gastric epithelial cells. Since TFF2 is a critical gastroprotective agent, PPARgamma may be involved in the gastric mucosal defense through regulating TFF2 expression in humans. PMID- 17118694 TI - Ntal/Lab/Lat2. AB - Non-T cell activation linker (NTAL)/linker for activation of B cells (LAB), now officially termed LAT2 (linker for activation of T cells 2) is a 25-30kDa transmembrane adaptor protein (TRAP) associated with glycolipid-enriched membrane fractions (GEMs; lipid rafts) in specific cell types of hematopoietic lineage. Tyrosine phosphorylation of NTAL/LAB/LAT2 is induced by FcvarepsilonRI aggregation and Kit dimerization in mast cells, FcgammaRI aggregation in monocytes, and BCR aggregation in B cells. NTAL/LAB/LAT2 is also expressed in resting NK cells but, unlike the related TRAP, LAT, not in resting T cells. As demonstrated in monocytes and B cells, phosphorylated NTAL/LAB/LAT2 recruits signaling molecules such as Grb2, Gab1 and c-Cbl into receptor-signaling complexes. Although gene knock out and knock down studies have indicated that NTAL/LAB/LAT2 may function as both a positive and negative regulator of mast cell activation, its precise role in the activation of these and other hematopoietic cells remains enigmatic. PMID- 17118695 TI - Urokinase-type plasminogen activator. AB - Urokinase-type plasminogen activator (uPA) is a serine protease involved in tissue remodeling and cell migration. At the gene level, the interplay between a complex enhancer, required for induced and basal transcription, and the minimal promoter finely tunes uPA expression. The active form of uPA is bound to its high affinity receptor on the cell surface, where specific inhibitors modulate its enzymatic activity. Such inhibitors also regulate the cell surface levels of uPA by triggering the internalization of the uPA-receptor-inhibitor complex. The role of uPA is not only linked to its action as an enzyme. In fact, the mere binding of uPA on the cell surface also brings about two events that broaden the spectrum of its biological functions: (1) a conformational change of the receptor, which, in turn, affects its interaction with other proteins; (2) a signal transduction which modulates the expression of apoptosis-related genes. Besides its applications as a thrombolytic agent and as a prognostic marker for tumors, uPA may provide the basis for other therapies, as the structure of the receptor binding domain of uPA has become a model for the design of anti-cancer molecules. PMID- 17118696 TI - Waste management - cytokines, growth factors and cachexia. AB - Muscle damage with a lack of regeneration, manifests itself in several life threatening diseases, including cancer cachexia, congestive heart failure, AIDS and sepsis. Often misdiagnosed as a condition simply of weight loss, cachexia is actually a highly complex metabolic disorder involving features of anorexia, anaemia, lipolysis and insulin resistance. A significant loss of lean body mass arises from such conditions, resulting in wasting of skeletal muscle. Unlike starvation, the weight loss seen in chronic illnesses arises equally from loss of muscle and of fat. The cachectic state is particularly problematic in cancer, typifying poor prognosis and often lowering responses to chemotherapy and radiation treatment. More than half of cancer patients suffer from cachexia, and strikingly, nearly one-third of cancer deaths are related to cachexia rather than the tumour burden. In considering this disorder, we are faced with a conundrum; how is it possible for uncontrolled growth to prevail in the tumour, in the face of unrestrained tissue loss in our muscles? Consistently, the catabolic state has been associated with a shift in the homeostatic balance between muscle synthesis and degradation mediated by the actions of growth factors and cytokines. Indeed, tumour necrosis factor-alpha (TNF-alpha) levels are raised in several animal models of cachectic muscle wasting, whereas the insulin-like growth factor (IGF) system acts potently to regulate muscle development, hypertrophy and maintenance. This concept of skeletal muscle homeostasis, often viewed as the net balance between two separate processes of protein synthesis and degradation has however changed. More recently, the view is that these two biochemical processes are not occurring independently of each other but in fact are finely co-ordinated by a web of intricate signalling networks. This review, therefore, aims to discuss data currently available regarding the mechanisms of degeneration and regeneration with specific emphasis on the potential and controversial cross-talk which may exist between anabolic growth factors (e.g. IGF-I) and catabolic cytokines (e.g. TNF-alpha). Also importantly, the potential impact at a cellular level of exercise, diet and age will be addressed. Finally, the ability to 'hi jack' signalling pathways traditionally believed to be for growth and survival or death will be reviewed. It is anticipated that such a review will highlight significant gaps in our knowledge of the cachectic state as well as provide caution with regards to therapeutics suggesting total block on inflammatory processes such as that associated with TNF-alpha action. PMID- 17118697 TI - Fourth International Workgroup on Genotoxicity testing: results of the in vivo Comet assay workgroup. AB - As part of the Fourth International Workshop on Genotoxicity Testing (IWGT), held 9-10 September 2005 in San Francisco, California, an expert working group on the Comet assay was convened to review and discuss some of the procedures and methods recommended in previous documents. Particular attention was directed at the in vivo rodent, alkaline (pH >13) version of the assay. The aim was to review those protocol areas which were unclear or which required more detail in order to produce a standardized protocol with maximum acceptability by international regulatory agencies. The areas covered were: number of dose levels required, cell isolation techniques, measures of cytotoxicity, scoring of comets (i.e., manually or by image analysis), and the need for historical negative/positive control data. It was decided that a single limit dose was not sufficient although the required number of dose levels was not stipulated. The method of isolating cells was thought not to have a qualitative effect on the assay but more data were needed before a conclusion could be drawn. Concurrent measures of cytotoxicity were required with histopathological examination of tissues for necrosis or apoptosis as the "Gold Standard". As for analysing the comets, the consensus was that image analysis was preferred but not required. Finally, the minimal number of studies required to generate a historical positive or negative control database was not defined; rather the emphasis was placed on demonstrating the stability of the negative/positive control data. It was also agreed that a minimum reporting standard would be developed which would be consistent with OECD in vivo genotoxicity test method guidelines. PMID- 17118698 TI - Genotoxic effects of potassium bromate on human peripheral lymphocytes in vitro. AB - The aim of this study was to investigate the genotoxic effects of potassium bromate, which is used as a bleaching agent in flour, on human peripheral blood lymphocytes in vitro by sister chromatid exchange (SCE), chromosomal aberrations (CA) and micronucleus (MN) tests, and also to determine whether it has any genotoxic potential for humans. Cells were treated with 400, 450, 500, 550 microg/ml concentrations of potassium bromate for 24 and 48 h. The SCE frequencies showed an increase after both treatment periods, however, the differences between the treated cells and the control groups were found to be statistically significant only for the 48-h treatment. In addition, potassium bromate statistically significantly induced CA after the 24-h and 48-h treatment periods. Strikingly, potassium bromate induced CA as much as the positive control, mitomycin-C (MMC). Furthermore, potassium bromate decreased both the cell proliferation index (PI) and the mitotic index (MI). Although micronucleus formation was induced by potassium bromate during the 24-h treatment period in a dose-dependent manner, only the doses 500 and 550 microg/ml yielded statistically significant results. In contrast, MN formation was significantly induced at all doses during the 48-h treatment period. These in vitro results provide important evidence about genotoxicity of potassium bromate on a human cell culture system. PMID- 17118699 TI - Synthesis, magnetic, spectral, and antimicrobial studies of Cu(II), Ni(II) Co(II), Fe(III), and UO2(II) complexes of a new Schiff base hydrazone derived from 7-chloro-4-hydrazinoquinoline. AB - A new hydrazone ligand, HL, was prepared by the reaction of 7-chloro-4 hydrazinoquinoline with o-hydroxybenzaldehyde. The ligand behaves as monoprotic bidentate. This was accounted for as the ligand contains a phenolic group and its hydrogen atom is reluctant to be replaced by a metal ion. The ligand reacted with Cu(II), Ni(II), Co(II), Fe(III), and UO2(II) ions to yield mononuclear complexes. In the case of Fe(III) ion two complexes, mono- and binuclear complexes, were obtained in the absence and presence of LiOH, respectively. Also, mixed ligand complexes were obtained from the reaction of the metal cations Cu(II), Ni(II) and Fe(III) with the ligand (HL) and 8-hydroxyquinoline (8-OHqu) in the presence of LiOH, in the molar ratio 1:1:1:1. It is clear that 8-OHqu behaves as monoprotic bidentate ligand in such mixed ligand complexes. The ligand, HL, and its metal complexes were characterized by elemental analyses, IR, UV-vis, mass, and 1H NMR spectra, as well as magnetic moment, conductance measurements, and thermal analyses. All complexes have octahedral configurations except Cu(II) complex which has an extra square-planar geometry, while Ni(II) mixed complex has also formed a tetrahedral configuration and UO2(II) complex which formed a favorable pentagonal biprymidial geometry. Magnetic moment of the binuclear Fe(III) complex is quite low compared to calculated value for two iron ions complex and thus shows antiferromagnetic interactions between the two adjacent ferric ions. The HL and metal complexes were tested against one stain Gram positive bacteria (Staphylococcus aureus), Gram negative bacteria (Escherichia coli), and fungi (Candida albicans). The tested compounds exhibited higher antibacterial acivities. PMID- 17118700 TI - Designing web services in health information systems: from process to application level. AB - Service-oriented architectures (SOAs) and web service technologies have been proposed to respond to some central interoperability challenges of heterogeneous health information systems (HIS). We propose a model which we are using to define services and solutions for healthcare applications from the requirements in the healthcare processes. Focusing on the transition from the process level of the model to the application level, we also present some central design considerations, which can be used to guide the design of service-based interoperability. We illustrate these aspects with examples from our current work from the service-enabled HIS. PMID- 17118701 TI - A notary archive model for secure preservation and distribution of electrically signed patient documents. AB - The healthcare industry is moving from paper-based documentation into the digital era. Electronic health records (EHR) are playing a major role in this development. Electronic health records will not only to be shared among a growing number of healthcare providers but they have also to be archived over long periods of time. The required life cycle depends of national regulations, but typically the preservation time of patient data varies between 20 and 100 years. Availability, integrity, confidentiality and non-repudiation of stored data over these lengthy preservation periods needs to be fully proven, both to preclude loss and also ensure the ability to read and understand content is maintained. This document describes a co-operative trusted notary archive (TNA) which receives granular health data from different EHR-systems, stores data together with associated meta-information for long periods and distributes granular EHR data objects. TNA communicates with EHR-systems and external users via archive request and distribution messages. TNA can store objects in XML-format and prove the non-repudiation and integrity of stored data with the help of event records, Time-stamps and archive e-signatures. PMID- 17118702 TI - Genetic distribution of group A human rotavirus types isolated in Gyunggi province of Korea, 1999-2002. AB - BACKGROUND: Human rotavirus genotypes G1-G4 and G9 are the major etiological agents of infantile gastroenteritis. G1 was the most prevalent in Korea during the 10-year period prior to 1997. However, between 1998 and 1999, G4 was the predominant type in Korea, as it was in other Asian countries. OBJECTIVES: The circulating pattern and genetic variability of group A human rotavirus in Gyunggi, Korea, 1999-2002, were examined in 189 stool specimens. STUDY DESIGN: Stool samples were collected from children with diarrhea, and group A human rotavirus type was determined using multiplex RT-PCR in those specimens found to be positive for rotavirus by ELISA. Each genotype was sequenced, and phylogenetic analysis was performed on the sequences. RESULT: We found significant variability from year to year in the prevalence of different G and P types of rotavirus. We also found relatively high prevalence rates for types normally considered to be uncommon. Furthermore, we found that the most prevalent combination of G and P types changed from year to year. Although the combination of G and P types changed every year, the sequence of G genotypes showed a high level of similarity (>97%) compared to those of strains from other Asian countries. CONCLUSION: We report the types of rotavirus circulating in Gyunggi province, Korea from 1999 to 2002. This information on rotavirus diversity has important implications for rotavirus vaccine efficacy and future vaccine development. PMID- 17118703 TI - Development and assessment of a rapid method to detect Escherichia coli O26, O111 and O157 in retail minced beef. AB - A molecular-based detection method was developed to detect Escherichia coli O26, O111 and O157 in minced (ground) beef samples. This method consists of an initial overnight enrichment in modified tryptone soya broth (mTSB) and novobiocin prior to DNA extraction and subsequent serogrouping using a triplex PCR. This method has a low limit of detection and results are available within 24 hours of receipt of samples. Once optimized, this rapid method was utilized to determine the prevalence of these E. coli serogroups in six hundred minced beef samples all of which were previously examined by immunomagnetic separation (IMS) and selective plating for E. coli O26 and O111. Using IMS, two E. coli O26 isolates were detected. No E. coli O111 were recovered. The multiplex PCR technique described here did not detect E. coli O111 nor O157 in any of the samples, however six minced beef samples were positive for E. coli O26 using our method, only two of these were previously detected by IMS and culture. Application of molecular methods are useful to support culture-based approaches thereby further contributing to risk reduction along the food chain. PMID- 17118704 TI - Warm-up or stretch as preparation for sprint performance? AB - Warm-up and stretching are widely used as techniques in preparation for intense physical activity, yet there is little information available to compare their effectiveness in relation to athletic performance. Fourteen elite Under-19 year old rugby league footballers undertook each of four preparation protocols (no preparation, stretching only, warm-up only, warm-up and stretching) in four successive testing sessions. Protocols were randomly allocated to players in a counterbalanced design so that each type of preparation occurred equally on each day of testing. During each session, athletes performed three solo sprint trials at maximum speed. Sprints were of 40-m distance and were electronically timed with wind speed and direction recorded. Preparation involving warm-up resulted in significantly faster sprint times compared to preparations having no warm-up, with a diminishing effect over the three trials. On the first trial, warm-up resulted in a mean advantage of 0.97 m over 40 m. Stretching resulted in a mean disadvantage of 0.18 m on the first trial, and no significant effect overall despite significant wind assistance. Warm-up was effective at improving immediate sprint performance, whereas an equivalent duration of lower limb stretching had no effect. PMID- 17118705 TI - Why Australia needs an effective national campaign to reduce coastal drowning. PMID- 17118706 TI - Contrast water immersion hastens plasma lactate decrease after intense anaerobic exercise. AB - The benefits of rapid recovery after intense exercise are widely recognised, and lactate elimination is one indicator of recovery rate. This study examined the effect of contrast (alternating hot and cold) water immersion (CWI) on the rate of plasma lactate decrease during recovery after intense anaerobic exercise. Eleven subjects on each of two occasions undertook four successive 30-s Wingate tests separated by 30-s rest periods. On each occasion, plasma lactate concentration during recovery was measured 5 min post-exercise and thereafter at 5 min intervals for 30 min. On one occasion (determined randomly), the subjects recovered passively (PR) on a recovery bed and, on the other, they alternated partial body immersion in hot (36 degrees C) and cold (12 degrees C) water baths. Plasma lactate concentrations were analysed by repeated measures analysis of variance and by fitting a linear regression model, allowing for both gender and recovery mode differences. The rate of decrease in plasma lactate concentration over the 30-min recovery period was significantly higher (p<0.001) in CWI; 0.28(+/-0.02) mmol L(-1) min(-1) (CWI) compared to 0.22(+/-0.02) mmol L(-1) min( 1) (PR). These values do not differ significantly between males and females. Contrast water immersion is a valid method of hastening plasma lactate decrease during recovery after intense anaerobic exercise for both males and females. An approximately 1.8 mmol L(-1) difference between the two conditions may be expected after 30 min. With differences among elite competitors as little as 1 2%, this reduction may be of practical significance. PMID- 17118707 TI - The STAT3 oncogene as a predictive marker of drug resistance. AB - Constitutive activation of STAT3 (signal transducer and activator of transcription) has been reported in several primary cancers and tumor cell lines where it induces cell transformation through a combined inhibition of apoptosis and cell-cycle activation. Several studies have suggested that STAT3 prevents cell-cycle arrest and cell death through upregulation of survival proteins and downregulation of tumor suppressors. As a consequence of anti-apoptotic and proliferative lesions, we propose that this oncogenic pathway is also involved in intrinsic drug resistance and that STAT3-expressing tumors are resistant to chemotherapeutic agents. If this hypothesis is correct, the detection of the activated form of this protein should help to define subsets of tumors that fail to respond to chemotherapy. Furthermore, interfering with the STAT3 oncogenic pathway might restore the sensitivity to anticancer drugs. PMID- 17118708 TI - New anthelmintics for livestock: the time is right. AB - The viability of small-ruminant enterprises is under threat in some locations because anthelmintic resistance continues to increase in prevalence and severity. Despite this, no new anthelmintic group has been developed for many years, and animal health companies seem unconvinced of the commercial justification for new compounds for the ruminant market. It is now argued, however, that market conditions are becoming favourable because of increases in multiple-drug resistance in nematodes of ruminants and other hosts and because few non-chemical options exist. A new anthelmintic is urgently needed and will find ready acceptance in an increasing market, and with better knowledge of resistance management, greater product longevity can be achieved. PMID- 17118709 TI - A biochemical comparison of a pharmaceutically licensed coagulation active plasma (Octaplas) with a universally applicable development product (Uniplas) and single donor FFPs subjected to methylene-blue dye and white-light treatment. AB - The strive for more standardised and highly efficacious products is one of the important mainstays in modern haemotherapy. Coagulation active plasma for transfusion is the product of choice when treating hereditary or acquired isolated or complex coagulopathies, when no specific concentrate is available. The aim of this study was to perform an extensive biochemical comparison of the pharmaceutically licensed coagulation active plasma named Octaplas with an identical, but universally applicable, development product (Uniplas, working title) and single-donor fresh-frozen plasma (FFP) units subjected to a medical device treatment using a combination of methylene-blue dye and subsequent white light exposure (MB plasma). Our study showed that there are differences in the biochemical characteristics between Octaplas and MB plasma, while Uniplas revealed the same quality as Octaplas. The variability of selected plasma proteins in the 20 individual MB plasma units tested was high compared to Octaplas/Uniplas. Beyond the reported decreased levels of protein S and plasmin inhibitor found in Octaplas/Uniplas, and the significant loss of fully functional fibrinogen in MB plasma and its impact on selected global coagulation parameters, the latter product additionally revealed several coagulation factor activities outside the ranges given for normal single-donor FFP. It is important for plasma prescribers to be aware of the major inherent differences between Octaplas and MB plasma. PMID- 17118710 TI - Endotoxin apheresis for sepsis. AB - The principle use of apheresis in the treatment of sepsis may be summarized as the removal of toxic substances and the restoration of normal organs function. It is ideal to control the early phases of inflammatory cascade when treating sepsis by removing microbial components, such as endotoxin or peptidoglycan. This review discusses endotoxin apheresis with particular emphasis on treatment using polymyxin B immobilized fiber columns (Toraymyxin) which are used widely in Japan for endotoxin removal therapy in patients with septic shock. Lixelle and CTR which have recently been shown to remove circulating bacterial components are also included in this review. PMID- 17118712 TI - Factors associated with employment in epilepsy patients. AB - OBJECTIVE: The aim of the study described here was to determine those variables associated with employment in patients followed at a level 4 epilepsy center. METHOD: A survey was sent to patients seen at the University of Florida Health Science Center/Jacksonville Comprehensive Epilepsy Program. RESULTS: Two hundred sixty-two eligible subjects constituted the study population. By univariate analysis, variables that distinguished employed patients included: younger age, Caucasian race, higher education and household income, not receiving disability benefits, currently studying, fewer seizure medications, having no other medical conditions that interfere with work, previous work experience, perceived importance of work for personal and financial reasons, and decreased fears of workplace discrimination. By logistic regression, higher annual family income, perceived importance of work for personal reasons, and decreased fears of workplace discrimination were the only variables independently associated with employment. CONCLUSION: Psychosocial factors such as a high self-perceived importance of work and decreased fears of workplace discrimination are significantly associated with employment in epilepsy patients. PMID- 17118713 TI - Autonomic dysfunction due to lead poisoning. AB - We present a case history of a 24 years old male who developed autonomic dysfunction, intestinal pseudo-obstruction and anemia due to lead poisoning. Concomitant recording of blood levels of lead and autonomic function showed a gradual decline in blood lead level (98.8 microg/dL at week 0, 56 microg/dL at week 6, and 40 microg/dL at week 52) and gradual improvement in autonomic functions. Decrease in blood lead levels with DMSA (Meso-2, 3-dimercaptosuccinic acid) therapy showed improvement in autonomic functions. At week 0, the patient had severe loss of autonomic tone and autonomic reactivity which improved at week 6. At the 52nd week, most of the autonomic parameters had normalized except for the persistence of mild loss of parasympathetic reactivity. PMID- 17118714 TI - A single HBsAg DNA vaccination in combination with electroporation elicits long term antibody responses in sheep. AB - Vaccines continue to be the most cost effective method to reduce the burden of disease in both human and animal health. However, there is a need to improve the duration of immunity following vaccination, since maintenance of protective levels of antibody in serum or the ability to rapidly respond upon re-exposure (memory) is critical if vaccines are to provide long-term protective immunity. The purpose of this experiment was to test the duration of antibody responses and the ability to generate anamnestic responses following a single immunization with a DNA vaccine encoding hepatitis B surface antigen (HBsAg) delivered by a variety of routes. Sheep immunized with the conventional HBsAg subunit vaccine (Engerix B) as well as sheep immunized with a HBsAg DNA vaccine, combined with electroporation, generated significant antibody responses that were sustained for 25 weeks after primary immunization. At 25 weeks, all experimental groups received a secondary immunization with the HBsAg subunit vaccine. Sheep that received a primary DNA immunization, in combination with electroporation, mounted an anamnestic response similar to the cohort immunized with the HBsAg subunit vaccine. In contrast, animals immunized with DNA vaccines administered without electroporation elicited no detectable memory response. The presence of immune memory was significantly correlated with the induction of a prolonged primary immune response. Thus, a single DNA vaccination, in combination with electroporation, approached the efficacy of the commercial subunit vaccine in the maintenance of long-term protective serum antibody titres and immune memory. PMID- 17118715 TI - Mammalian single-strand break repair: mechanisms and links with chromatin. AB - Thousands of cellular single-strand breaks (SSBs) arise in cells each day, from attack of deoxyribose and DNA bases by reactive oxygen species and other electrophilic molecules, and from the intrinsic instability of DNA. If not repaired, SSBs can disrupt transcription and replication and can be converted into potentially clastogenic and/or lethal DNA double-strand breaks. Here, I present an updated model for the repair of SSBs, and speculate on the possible impact of chromatin structure and remodelling on single-strand break repair (SSBR) processes. PMID- 17118716 TI - A unique error signature for human DNA polymerase nu. AB - Human DNA polymerase nu (pol nu) is one of three A family polymerases conserved in vertebrates. Although its biological functions are unknown, pol nu has been implicated in DNA repair and in translesion DNA synthesis (TLS). Pol nu lacks intrinsic exonucleolytic proofreading activity and discriminates poorly against misinsertion of dNTP opposite template thymine or guanine, implying that it should copy DNA with low base substitution fidelity. To test this prediction and to comprehensively examine pol nu DNA synthesis fidelity as a clue to its function, here we describe human pol nu error rates for all 12 single base-base mismatches and for insertion and deletion errors during synthesis to copy the lacZ alpha-complementation sequence in M13mp2 DNA. Pol nu copies this DNA with average single-base insertion and deletion error rates of 7 x 10(-5) and 17 x 10( 5), respectively. This accuracy is comparable to that of replicative polymerases in the B family, lower than that of its A family homolog, human pol gamma, and much higher than that of Y family TLS polymerases. In contrast, the average single-base substitution error rate of human pol nu is 3.5 x 10(-3), which is inaccurate compared to the replicative polymerases and comparable to Y family polymerases. Interestingly, the vast majority of errors made by pol nu reflect stable misincorporation of dTMP opposite template G, at average rates that are much higher than for homologous A family members. This pol nu error is especially prevalent in sequence contexts wherein the template G is preceded by a C-G or G-C base pair, where error rates can exceed 10%. Amino acid sequence alignments based on the structures of more accurate A family polymerases suggest substantial differences in the O-helix of pol nu that could contribute to this unique error signature. PMID- 17118717 TI - XRCC1 interactions with base excision repair DNA intermediates. AB - Abasic (AP) sites in DNA arise either spontaneously, or through glycosylase catalyzed excision of damaged bases. Their removal by the base excision repair (BER) pathway avoids their mutagenic and cytotoxic consequences. XRCC1 coordinates and facilitates single-strand break (SSB) repair and BER in mammalian cells. We report that XRCC1, through its NTD and BRCT1 domains, has affinity for several DNA intermediates in BER. As shown by its capacity to form a covalent complex via Schiff base, XRCC1 binds AP sites. APE1 suppresses binding of XRCC1 to unincised AP sites however, affinity was higher when the DNA carried an AP lyase- or APE1-incised AP site. The AP site binding capacity of XRCC1 is enhanced by the presence of strand interruptions in the opposite strand. Binding of XRCC1 to BER DNA intermediates could play an important role to warrant the accurate repair of damaged bases, AP sites or SSBs, in particular in the context of clustered DNA damage. PMID- 17118718 TI - A comparative study of proteolysis methods for the measurement of 3-nitrotyrosine residues: enzymatic digestion versus hydrochloric acid-mediated hydrolysis. AB - A common approach for the quantification of 3-nitrotyrosine (NY) in routine analyses relies on the cleavage of peptide bonds in order to release the free amino acids from proteins in tissues or fluids. NY is usually monitored by either GC-MS(/MS) or LC-MS/MS techniques. Various proteolysis methods have been employed to combine digestion efficiency with prevention of artifactual nitration of tyrosine. However, so far, no study was designed to compare the HCl-based hydrolysis method with enzymatic digestion in terms of reliability for the measurement of NY. The present work addresses the digestion efficiency of BSA using either 6M HCl, pronase E or a cocktail of enzymes (pepsin, pronase E, aminopeptidase, prolidase) developed in our laboratory. The HCl-based hydrolysis leads to a digestion yield of 95%, while 25 and 75% are achieved with pronase E and the cocktail of enzymes, respectively. These methods were compared in terms of NY measurement and the results indicate that a prior reduction of the disulfide bonds ensures a reliable quantification of NY. We additionally show that the enzyme efficacy is not altered when the digestion is carried out in the presence of BSA with a high content of NY. PMID- 17118719 TI - Liquid chromatographic determination of irbesartan in human plasma. AB - A simple and sensitive method was developed for determination of irbesartan by liquid chromatography with fluorescence detection. Irbesartan and losartan (I.S.) in human plasma were extracted using diethyl ether:dichloromethane (7:3, v/v) followed by back extraction with 0.05 M sodium hydroxide. Neutralized samples were analyzed using 0.01 M potassium dihydrogen phosphate buffer (containing 0.07% triethylamine as peak modifier, pH was adjusted with orthophosphoric acid to pH 3.0) and acetonitrile (66:34, v/v). Chromatographic separation was achieved on an ODS-C-18 column (100 mm x 4.6 mm i.d., particle size 5 microm) using isocratic elution (at flow rate 1.25 ml/min). The peak was detected using a fluorescence detector set at Ex 259 nm and Em 385 nm, and the total time for a chromatographic separation was approximately 13 min. The validated quantitation ranges of this method were 15-4000 ng/ml with coefficients of variation between 0.75 and 12.53%. Mean recoveries were 73.3-77.1% with coefficients of variation of 3.7-6.3%. The between- and within-batch precision were 0.4-2.2% and 0.9-6.2%, respectively. The between- and within-batch relative errors (bias) were (-5.5) to 0.9% and (-0.6) to 6.9%, respectively. Stability of irbesartan in plasma was >89%, with no evidence of degradation during sample processing and 60 days storage in a deep freezer at -70 degrees C. This validated method is sensitive and simple with between-batch precision of <3% and can be used for pharmacokinetic studies. PMID- 17118720 TI - SELDI-TOF mass spectra: a view on sources of variation. AB - Adequate interpretation of mass spectrometry data can yield valuable biomarkers. However, spectrum interpretation is a complicated task. This paper reviews the various factors that determine a sample's spectrum and demonstrates the role of these factors in the interpretation process. We derive a simulation model that adequately predicts the expected spectrum based on known sample content and, in the reverse mode, obtain an analysis model that adequately fits an observed spectrum based on the hypothesized sources of variation. PMID- 17118721 TI - Characterization of flavonoids in the traditional Chinese herbal medicine Huangqin by liquid chromatography coupled with electrospray ionization mass spectrometry. AB - The root of Scutellaria baicalensis, called Huangqin in Chinese, is one of the most commonly used traditional Chinese medicines for the treatment of hepatitis, tumors, diarrhea, and inflammatory diseases. The major chemical constituents of Huangqin are flavonoids. In the present paper, HPLC-DAD-ESI-MS(n) was used to analyze flavonoids in the roots of S. baicalensis. A total of 26 flavonoids were identified or tentatively characterized, including 5 C-glycosides, 12 O glycosides, and 9 free aglycones. Two C-glycosides, apigenin-6-C-glucyl-8-C arabinoside and chrysin-6,8-di-C-glucoside, together with some O-glycosides, are reported from S. baicalensis for the first time. This method is simple, reliable and sensitive, and could be used for the quality control of Huangqin and its related preparations. PMID- 17118722 TI - Quantification of urinary zwitterionic organic acids using weak-anion exchange chromatography with tandem MS detection. AB - A rapid and accurate quantitative method was developed and validated for the analysis of four urinary organic acids with nitrogen containing functional groups, formiminoglutamic acid (FIGLU), pyroglutamic acid (PYRGLU), 5 hydroxyindoleacetic acid (5-HIAA), and 2-methylhippuric acid (2-METHIP) by liquid chromatography tandem mass spectrometry (LC/MS/MS). The chromatography was developed using a weak anion-exchange amino column that provided mixed-mode retention of the analytes. The elution gradient relied on changes in mobile phase pH over a concave gradient, without the use of counter-ions or concentrated salt buffers. A simple sample preparation was used, only requiring the dilution of urine prior to instrumental analysis. The method was validated based on linearity (r2>or=0.995), accuracy (85-115%), precision (C.V.<12%), sample preparation stability (2U model yielded a DeltaG of approximately -10 kcal/mol. Our results suggest that 26-71(A3) FIS can form a stable dimeric structure despite lacking the N- and C-terminus of native FIS. PMID- 17118727 TI - The effect of dextran on subunit exchange of the molecular chaperone alphaA crystallin. AB - Alpha-crystallin, a member of small heat shock protein (sHsp) family, is comprised of alphaA and alphaB subunits and acts as a molecular chaperone by interacting with unfolding proteins to prevent their aggregation. The alphaA crystallin homopolymer consists of 30-40 subunits that are undergoing dynamic exchange. In vivo, alpha-crystallin elicits its chaperone action in a crowded cellular environment (e.g. in the lens). In vitro, inert molecular crowding agents (e.g. dextran) are often used to mimic crowded conditions. In this study, it was found that alpha-crystallin and alphaA-crystallin are poorer chaperones in the presence of dextran. Using fluorescence resonance energy transfer, it is shown that the alphaA-crystallin subunit exchange rate strongly increases with temperature. Binding of reduced ovotransferrin to alphaA-crystallin markedly decreases the rate of subunit exchange, as does the presence of dextran. In addition, in the presence of dextran the effect of reduced ovotransferrin on decreasing the rate of subunit exchange of alphaA-crystallin is greater than in the absence of dextran. Under the conditions of molecular crowding, the alphaA crystallin subunit exchange rate is not temperature-dependent. In the absence of dextran, the exchange rate of alphaA-crystallin subunits correlates with its chaperone efficiency, i.e. the chaperone ability of alphaA-crystallin increases with temperature. However in the presence of dextran, the temperature dependence of the chaperone ability of alphaA-crystallin is eliminated. PMID- 17118728 TI - IPCS framework for analyzing the relevance of a cancer mode of action for humans. AB - The use of structured frameworks can be invaluable in promoting harmonization in the assessment of chemical risk. IPCS has therefore updated and extended its mode of action (MOA) framework for cancer to address the issue of human relevance of a carcinogenic response observed in an experimental study. The first stage is to determine whether it is possible to establish an MOA. This comprises a series of key events along the causal pathway to cancer, identified using a weight-of evidence approach based on the Bradford Hill criteria. The key events are then compared first qualitatively and then quantitatively between the experimental animals and humans. Finally, a clear statement of confidence, analysis, and implications is produced. The IPCS human relevance framework for cancer provides an analytical tool to enable the transparent evaluation of the data, identification of key data gaps, and structured presentation of information that would be of value in the further risk assessment of the compound, even if relevancy cannot be excluded. This might include data on the shape of the dose response curve, identification of any thresholds and recognition of potentially susceptible subgroups, for example, the basis of genetic or life-stage differences. PMID- 17118729 TI - Thiazopyr and thyroid disruption: case study within the context of the 2006 IPCS Human Relevance Framework for analysis of a cancer mode of action. AB - Thiazopyr increases the incidence of male rat thyroid follicular-cell tumors; however, it is not carcinogenic in mice. Thiazopyr is not genotoxic. Thiazopyr exerts its carcinogenic effect on the rat thyroid gland secondary to enhanced metabolism of thyroxin leading to hormone imbalance. The relevance of these rat tumors to human health was assessed by using the 2006 IPCS Human Relevance Framework. The postulated rodent tumor mode of action was tested against the Bradford Hill criteria and was found to satisfy the conditions of dose and temporal concordance, biological plausibility, coherence, strength, consistency, and specificity that fits with a well-established mode of action for thyroid follicular-cell tumors. Although the postulated mode of action could theoretically operate in humans, marked quantitative differences in the inherent susceptibility for neoplasia to thyroid hormone imbalance in rats allows for the conclusion that thiazopyr does not pose a carcinogenic hazard to humans. PMID- 17118730 TI - 4-Aminobiphenyl and DNA reactivity: case study within the context of the 2006 IPCS Human Relevance Framework for Analysis of a cancer mode of action for humans. AB - The IPCS Human Relevance Framework was evaluated for a DNA-reactive (genotoxic) carcinogen, 4-aminobiphenyl, based on a wealth of data in animals and humans. The mode of action involves metabolic activation by N-hydroxylation, followed by N esterification leading to the formation of a reactive electrophile, which binds covalently to DNA, principally to deoxyguanosine, leading to an increased rate of DNA mutations and ultimately to the development of cancer. In humans and dogs, the urinary bladder urothelium is the target organ, whereas in mice it is the bladder and liver; in other species, other tissues can be involved. Differences in organ specificity are thought to be due to differences in metabolic activation versus inactivation. Based on qualitative and quantitative considerations, the mode of action is possible in humans. Other biological processes, such as toxicity and regenerative proliferation, can significantly influence the dose response of 4-aminobiphenyl-induced tumors. Based on the IPCS Human Relevance Framework, 4-aminobiphenyl would be predicted to be a carcinogen in humans, and this is corroborated by extensive epidemiologic evidence. The IPCA Human Relevance Framework is useful in evaluating DNA-reactive carcinogens. PMID- 17118731 TI - Formaldehyde and glutaraldehyde and nasal cytotoxicity: case study within the context of the 2006 IPCS Human Framework for the Analysis of a cancer mode of action for humans. AB - Formaldehyde and glutaraldehyde cause toxicity to the nasal epithelium of rats and mice upon inhalation. In addition, formaldehyde above certain concentrations induces dose-related increases in nasal tumors in rats and mice, but glutaraldehyde does not. Using the 2006 IPCS human framework for the analysis of cancer mode of action (MOA), an MOA for formaldehyde was formulated and its relevance was tested against the properties of the noncarcinogenic glutaraldehyde. These compounds produce similar patterns of response in histopathology and in genotoxicity tests (although formaldehyde has been much more extensively tested studied). The MOA is based on the induction of sustained cytotoxicity and reparative cell proliferation induced by formaldehyde at concentrations that also induce nasal tumors upon long-term exposure. Data on dose dependency and temporal relationships of key events are consistent with this MOA. While a genotoxic MOA can never be ruled out for a compound that is clearly genotoxic, at least in vitro, the nongenotoxic properties fundamental to the proposed MOA can explain the neoplastic response in the nose and may be more informative than genotoxicity in risk assessment. It is not yet fully explained why glutaraldehyde remains noncarcinogenic upon inhalation, but its greater inherent toxicity may be a key factor. The dual aldehyde functions in glutaraldehyde are likely to produce damage resulting in fewer kinetic possibilities (particularly for proteins involved in differentiation control) and lower potential for repair (nucleic acids) than would be the case for formaldehyde. While there have been few studies of possible glutaraldehyde associated cancer, the evidence that formaldehyde is a human carcinogen is strong for nasopharyngeal cancers, although less so for sinonasal cancers. This apparent discrepancy could be due in part to the classification of human nasal tumors with tumors of the sinuses, which would receive much less exposure to inhaled formaldehyde. Evaluation of the human relevance of the proposed MOA of formaldehyde in rodents is restricted by human data limitations, although the key events are plausible. It is clear that the human relevance of the formaldehyde MOA in rodents cannot be excluded on either kinetic or dynamic grounds. PMID- 17118732 TI - Gestational programming of offspring obesity/hypertension. AB - The intrauterine milieu impacts fetal growth directly during gestation. It is now clear, however, that postnatal phenotype is also influenced by prenatal conditions. A variety of disorders in the adult have been linked to fetal size at birth; these include glucose intolerance, cardiovascular disease, and the subjects of this review, obesity and hypertension. We will review recent data regarding these associations and the pathophysiologic mechanisms underlying them in humans as well as in animal models. PMID- 17118733 TI - The pathophysiology of trauma in pregnancy: a review. AB - Emergency care of the pregnant patient with trauma presents a unique set of circumstances and challenges to physicians. Pregnancy causes anatomic and physiologic changes involving nearly every organ system in the body, making treatment of the pregnant trauma patient difficult. The other factors that make treatment complex are fear of harming the fetus, upsetting the patient, and/or lack of experience. The possibility of pregnancy should be considered in all women of reproductive age with trauma. A profound understanding of the pathophysiology of the pregnant trauma patient might aid in dealing with this complex problem. PMID- 17118734 TI - A role of the anti-angiogenic factor sVEGFR-1 in the 'mirror syndrome' (Ballantyne's syndrome). AB - BACKGROUND: 'Mirror syndrome' (Ballantyne's syndrome) refers to the association of fetal hydrops with placentomegaly and severe maternal edema. Preeclampsia occurs in approximately 50% of these cases. Soluble vascular endothelial growth factor receptor-1 (sVEGFR-1), an anti-angiogenic factor, has been implicated in the pathophysiology of preeclampsia (PE). OBJECTIVE: The objective of this study was to determine if the maternal plasma concentration of sVEGFR-1 is elevated in patients with mirror syndrome. STUDY DESIGN: This case-control study included patients with uncomplicated pregnancies (n = 40) and those with mirror syndrome (n = 4) matched for gestational age. Mirror syndrome was defined as fetal hydrops and severe maternal edema. Maternal plasma sVEGFR-1 concentrations were determined using specific enzyme-linked immunosorbent assays. Immunohistochemistry of sVEGFR-1 on villous trophoblasts was also performed in samples from one patient with mirror syndrome and compared with those from a patient with spontaneous preterm delivery matched for gestational age. Non parametric statistics were used for analysis (p < 0.05). RESULTS: (1) The median maternal plasma concentration of sVEGFR-1 was significantly higher in patients with mirror syndrome than in the control group (median: 3974 pg/mL, range: 3083 10 780 vs. median: 824 pg/mL, range: 260-4712, respectively; p < 0.001). (2) All patients with mirror syndrome had sVEGFR-1 concentrations above the 95th percentile for gestational age. Syncytiotrophoblast, especially syncytial knots, showed strong staining with antibodies against sVEGFR-1 in placental samples from the patient with mirror syndrome, but not in those from the patient with spontaneous preterm delivery. CONCLUSION: High maternal plasma concentrations of sVEGFR-1 were observed in mirror syndrome. We propose that this anti-angiogenic factor may participate in the pathophysiology of this syndrome. Thus, maternal plasma determination of sVEGFR-1 may help to identify the hydropic fetus that places the mother at risk for preeclampsia. PMID- 17118735 TI - Endocrine regulation in asymmetric intrauterine fetal growth retardation. AB - OBJECTIVE: The ponderal index (PI) is a widely accepted measure of disproportionate growth or asymmetrical growth retardation by pediatricians worldwide. Identification of disproportionately grown small for gestational age (SGA) neonates by using the ponderal index as a measure of the nutritional status at birth, is important because they constitute a high-risk group among SGA neonates. Poor nutritional status of the mother could have a direct effect on the organs of the developing fetus and/or affect the endocrine milieu in the maternal feto-placental unit resulting in an increased incidence of intrauterine growth retarded (IUGR)/SGA births. IUGR is a significant risk factor for adult disease. In this study, we have investigated the endocrine adaptation by the fetus to overcome the growth disadvantage caused due to poor nutritional status of the mother. MATERIALS AND METHODS: We examined the quantitative variations in hormonal and growth factor profiles in paired maternal and cord blood samples obtained from mothers and their neonates who were classified based on their growth status into SGA and appropriate for gestational age (AGA). RESULTS: (1) A total of 24.7% neonates had a PI < 2, indicating a high incidence of asymmetric IUGR in the population studied. (2) Anthropometric parameters measured in the mothers indicate that the mothers giving birth to neonates with a PI < 2 had poor nutritional status, both prior to and during pregnancy. (3) We observed increased levels of placental lactogen and prolactin and decreased levels of insulin in the cord blood of neonates with PI < 2, while lower levels of insulin-like growth factor 1 (IGF-1) and higher levels of epidermal growth factor (EGF) were observed in their mothers. CONCLUSION: Poor maternal nutritional status results in fetal adaptation to a growth restricted environment via the modulation of the pituitary thyroid axis thereby altering the endocrine milieu, thus affecting fetal growth. PMID- 17118736 TI - MCI-186 administered to the maternal circulation inhibits fetal brain injury resulting from total umbilical cord occlusion in the chronically instrumented fetal lamb. AB - OBJECTIVE: To evaluate the transplacental effects of MCI-186 (edaravone), a potent hydroxyl radical scavenger, administered to the maternal circulation to inhibit fetal brain injury caused by umbilical cord occlusion. METHODS: Nine chronically instrumented lambs were prepared. In three cases, 10-min persistent total umbilical cord occlusion (group A) was performed. Another three cases underwent occlusion and were administered 60 mg of MCI-186 through the maternal femoral vein prior to the end of occlusion (group B). The remaining three cases underwent sham operation (group C). On day 3 after insult, fetal brains were extirpated. Paraffin-embedded brain tissue sections were stained with hematoxylin and eosin, Bodian, Kluver-Barrera, and TUNEL. Neuronal cellular damage was evaluated by two pathologists blinded to the experimental conditions. RESULTS: Group A displayed numerous cells with eosinophilic condensation of nuclear chromatin and proliferation of microglia in the hippocampus and basal ganglia. TUNEL-positive cells were observed in the periventricular area. Group B showed microglial proliferations, but no marked changes. No pathological changes were apparent in group C. CONCLUSIONS: MCI-186 administered to the maternal circulation could inhibit fetal brain injury resulting from hypoxia-reperfusion induced by umbilical cord occlusion. PMID- 17118737 TI - Differences in amniotic fluid patterns and fetal biometric parameters in third trimester pregnancies with and without diabetes. AB - OBJECTIVE: The amniotic fluid index (AFI) has been increasingly used in the assessment of fetal well-being. We conducted the study to evaluate and compare the amniotic fluid index in third trimester normal and diabetic human pregnancy and to assess the correlation between the AFI and the fetal biometric parameters. METHODS: Real-time ultrasound was performed to evaluate the AFI (four-quadrant technique), and to measure the biparietal diameter, head circumference, abdominal circumference, and femur length in 225 normal and 120 diabetic pregnant women from 27 to 42 weeks of gestation. Each patient was studied only once. RESULTS: AFI in normal pregnancies was less than that in diabetic pregnancies throughout the gestational ages studied (27-42 weeks). In normal pregnancy, the mean AFI was 14.0 cm at 27 weeks and decreased to 11.4 cm at 42 weeks (r = 0.25, p = 0.0005), whereas in diabetic pregnancies, the values remained stable throughout the gestational ages studied. There exist significant differences in AFI, estimated fetal weight, estimated fetal weight %, abdominal circumference, abdominal circumference %, and head circumference to abdominal circumference ratio between the two groups. In both normal and diabetic pregnancies, there is a positive correlation between the AFI and the percentile of abdominal circumference (p < 0.0001), and between the AFI and the percentile of estimated fetal weight (p < 0.0001). CONCLUSION: This study provides gestational age-specific values of the AFI in normal and diabetic pregnancies. Diabetic pregnancy has greater AFI values than normal pregnancy between 27 and 42 weeks. The AFI correlates to the percentile of the estimated fetal weight and the abdominal circumference in both groups, suggesting that there may be a relationship between increased AFI and large for gestational age fetus independent of diabetes. PMID- 17118738 TI - Single- versus double-layer uterine incision closure and uterine rupture. AB - OBJECTIVE: To evaluate whether closure of the uterine incision with one or two layers changes uterine rupture or vaginal birth after cesarean section (VBAC) success rates. METHODS: Subjects with one previous cesarean section by documented transverse uterine incision that attempted VBAC were identified. Exclusion criteria included lack of documentation of the type of closure of the previous uterine incision, multiple gestation, more than one previous cesarean section, and previous scar other than low transverse. Uterine rupture and VBAC success rates were compared between those with single-layer and double-layer uterine closure. Time interval between deliveries, birth weight, body mass index (BMI), and history of previous VBAC were evaluated as possible confounders. RESULTS: Of 948 subjects identified, 913 had double-layer closure and 35 had single-layer closure. The uterine rupture rate was significantly higher in the single-layer closure group (8.6% vs. 1.3%, p = 0.015). This finding persisted when controlling for previous VBAC, induction, birth weight >4000 g, delivery interval >19 months, and BMI >29 (OR 8.01, 95% CI 1.96-32.79). There was no difference in VBAC success rate (74.3% vs. 77%, p = 0.685). CONCLUSION: Single-layer uterine closure may be more likely to result in uterine rupture. PMID- 17118739 TI - Matching risk and resources in high-risk pregnancies. AB - OBJECTIVE: To evaluate the joint impact of pregnancy risk and the timing of referral of high-risk pregnancies from obstetricians to maternal fetal medicine (MFM) sub-specialists on gestational age (GA) at delivery. METHODS: For the period 1992-2002, 2567 consecutive deliveries from pregnancies of at least 23 weeks gestational age (GA) from a community-level sub-specialty perinatal center were studied. A multiple regression model was developed specifying the impact of various risk factors and referral timing. RESULTS: Prior pregnancy risk was inversely related to GA at birth. Referral timing, operationalized as a continuous variable, did not have a significant additive impact on GA at birth, but several dummy-variable interaction effects combining risk factors and referral before 20 weeks as a dichotomy were significant. CONCLUSION: There are identifiable risks that occur either before the pregnancy or early into it that should lead to early referral to a sub-specialist because of their impact on GA at birth. Early referral is an important tactic in a larger preterm prevention strategy, but it needs to be embedded in a broader maternal-fetal health initiative in which both generalists and sub-specialists play important roles. PMID- 17118740 TI - Adverse effects of thyroid dysfunction on pregnancy and pregnancy outcome: epidemiologic study in Slovenia. AB - OBJECTIVE: To evaluate the consequences of maternal thyroid dysfunction for pregnancy outcome. METHODS: A retrospective analysis involving all pregnant women who delivered in Slovenia in the 1997-1999 triennium; those having a medical history of thyroid dysfunction and/or taking thyroid medications were allotted to the study group (n = 748) and the remaining ones to the control group (n = 52 253). RESULTS: Significantly higher incidences of infertility (5.5% vs. 3.7%, p < 0.05), menstrual cycle irregularities (3.2% vs. 1.9%, p < 0.05), hypertensive disorders (7.0% vs. 4.2%, p < 0.05), threatened preterm delivery (9.1% vs. 5.6%, p < 0.001), and delivery before 32 weeks (2.7% vs. 1.5%, p < 0.05) were found in the study than in the control group. There were no significant differences in the incidences of miscarriage, non-gestational diabetes mellitus, proteinuria, hyperemesis, intrahepatic cholestasis of pregnancy, intrauterine growth restriction, placental abruption, preterm delivery, small for gestational age newborns (SGA), and stillbirths. CONCLUSIONS: This is the first study to evaluate the incidence of thyroid dysfunction for the whole population of pregnant women in Slovenia using a retrospective analysis. Thyroid dysfunction adversely affects pregnancy and pregnancy outcome but to a lesser extent than presented in previous studies. An evaluation of thyroid function in the women who experience menstrual cycle irregularities, infertility, and complications during pregnancy, labor and delivery would be advisable. PMID- 17118741 TI - Etiological risk factors for brachial plexus palsy. AB - AIM: To investigate risk factors for brachial plexus palsy in newborns. We analyzed 45 544 live-born children, born over a nine-year period from January 1, 1996 to December 31, 2004. METHODS: The analysis was retrospective and based on the medical documentation of the Clinic for Gynecology and Obstetrics, Clinic for Neurology, and Clinic for Physical Medicine and Rehabilitation of the University Clinical Center Tuzla. We compared study and control groups of newborns. Rates among groups were compared using Chi-square, with significance at p < 0.05, and with significance at p < 0.01. RESULTS: Examining epidemiological characteristics, 86 newborns with brachial plexus palsy had been recorded, thus, the prevalence was 1.86 per 1000 live-born children. Analyzing maternal and neonatal factors, and the labor pattern itself, it was found that the highest factors of risk for brachial plexus injury were birth weight of over 4000 g, a precipitous second stage of labor (<15 minutes), and vacuum-extractor assisted labor. Brachial plexus palsy was more frequent when the mothers were overweight, with a body mass index >or=29 kg/m2. None of the parturient women, whose newborns were diagnosed with brachial plexus palsy, had external conjugate diameter <18 cm. Newborns delivered vaginally were not diagnosed with a higher frequency of brachial plexus palsy when compared to newborns who were delivered by cesarean section, but newborns who were vaginal breech-delivered were diagnosed to have a higher incidence of brachial plexus palsy. Newborns whose mothers were older than 35 years were diagnosed to have brachial plexus palsy more frequently, but a statistically significant difference between primiparas and multiparas was not found. A total of 39 newborns (45.2%) were diagnosed with a fracture of the clavicle, which was the most frequently combined damage with brachial plexus injury. Forty-two newborns (48.8%) had an Apgar score of G, c.524G > A, c.528C > G, c.529_546del18, c.536A > G, c.736A > G, c.743G > A, c.770_771del2, c.772G > T, c.814G > A, c.949delC, c.782+1G > A, and c.919+1G > C. Only a double heterozygote for c.370T > G and c.529_546del18 was found. All the other mutations were heterozygous. No homozygote was found. 524G > A was the most common mutation, accounting for 18.75 percent (3/16). c.370T > G could not be found in the whole somatic mutation section of IARC TP 53 mutation database (2004 version). c.528C > G, c.949delC, c.782+1G > A, and c.919+1G > C could not be found in colorectal carcinomas reported in the somatic mutation section of IARC TP 53 mutation database (2004 version). The frequency (45.83 percent) of p53 gene mutations in > or = 60 years old patients was higher than that (16.67 percent) in < 60 year old patients (P < 0.05). The frequency (47.83 percent) of p53 gene mutations in rectal carcinoma was higher than that (16 percent) in colonic carcinoma (P < 0.025). PMID- 17118780 TI - Current update of cytokines in pancreatic cancer: pathogenic mechanisms, clinical indication, and therapeutic values. AB - Pancreatic cancer is the fourth leading cause of cancer death with a 5-year survival less than 5 percent despite rigorous interventions. This largely is due to its late presentation, aggressive metastasis, and a lack of effective adjuvant therapies. Cytokines have been studied in many tumor types, where they have been shown to be an important influence in cancer cell behavior and to have potential as tumor markers, therapeutic targets, or as treatments themselves. Recently, the roles cytokines play in pancreatic cancer have become the subject of intense investigation. However, the story is complicated, largely because of the pleiotropic and overlapping nature of cytokine functions. This article attempts to provide a focused review of recent discoveries in this area by organizing the material along the pathophysiologic tasks a cancer cell must accomplish to achieve malignancy. We examined how cytokines act to create a microenvironment conducive to tumor cell survival and growth, discussed how cytokines affect proliferation, invasion, metastasis, and apoptosis in pancreatic cancer cells, and we summarized how this knowledge has been applied either to target cytokines or use them therapeutically. PMID- 17118781 TI - A systematic review on drug interactions in oncology. AB - OBJECTIVES: To systematically review drug interactions in oncology. METHODS: We searched PubMed for eligible articles and online databases abstracts of major oncology meetings from 2002 to 2005. RESULTS: One study reported on the frequency of drug interactions. Interactions between chemotherapy and nonchemotherapy agents have been reported mostly in small clinical trials and case series. Interactions between chemotherapic agents have been reported mostly in Phase I studies. Few studies described fatal outcomes of drug interactions in cancer patients. CONCLUSION: Drug interactions comprise an important issue in oncology, but very limited data exist on their frequency and clinical consequences. PMID- 17118782 TI - Could BEACOPP be the new standard for the treatment of advanced Hodgkin's lymphoma (HL)? AB - In 1992, the German Hodgkin Study Group (GHSG) developed the BEACOPP regimen for further improving the outcome of patients with advanced Hodgkin's lymphoma (HL). Since then, BEACOPP has been introduced in 3 different prospective randomized clinical trials of the GHSG to find an equilibrium between maximal efficacy and least toxicity with the BEACOPP principle for the treatment of advanced stage HL. In the HD9 trial of the GHSG, with 1,186 patients, after a median observation time (mot) of 7 years, the rates for freedom from treatment failure (FFTF) are 85 percent, and for overall survival (OS) 90 percent for dose-escalated BEACOPP, and for COPP/ABVD (C/ABVD comparable to ABVD) the rate for FFTF is 67 percent, and for OS it is 79 percent. These superior BEACOPP results are obtained inspite of a higher rate of secondary AML/MDS in the esc. BEACOPP arm. The number of toxic deaths during treatment, however, was lower for esc. BEACOPP (1.6 percent) than for C/ABVD (1.8 percent). The majority of patients were treated in outpatient setting, in a multicenter study with more than 400 centers, including 120 private doctors, in Germany and 9 other European countries. The reduce acute and longterm toxicity, the GHSG started in the consecutive studies HD12 and HD15 for advanced stage HL to de-escalate BEACOPP by reducing the number of escalated BEACOPP cycles and by applying the baseline-dose BEACOPP, a time-dense regimen, called BEACOPP-14. The excellent results obtained with the BEACOPP principle challenge the seemingly global consensus that ABVD is the gold standard treatment strategy for advanced stage HL. PMID- 17118783 TI - Alemtuzumab in CLL and other lymphoid neoplasms. AB - The recent success of monoclonal antibodies in the treatment of various hematological and nonhematological cancers is the result of several decades of research in immune therapy of cancer. The identification of cancer-specific surface markers has led to the development of numerous monoclonal antibodies directed at these antigens, which have been associated with variable success in treating patients with different malignancies. Alemtuzumab, one such monoclonal antibody, is a humanized antibody directed against CD52. The target antigen is a small glycosylphosphatidylinositol (GPI)-anchored glycoprotein that is highly expressed on normal T- and B-lymphocytes and on a large proportion of malignant lymphoid cells, but not on hematopoietic progenitor cells. A number of clinical trials have demonstrated the clinical activity of alemtuzumab in chronic lymphocytic leukemia (CLL), T-cell malignancies such as T-prolymphocytic leukemia (T-PLL) and cutaneous T-cell lymphoma (CTCL), and have examined its role as an immunosuppressive agent in transplantation and for the treatment of autoimmune disorders. Effective antibiotic prophylaxis can limit the incidence of infections, which are the major side effect associated with the profound lymphopenia occurring as a result of treatment with this agent. PMID- 17118784 TI - Proceedings of the 10th Swiss Receptor Workshop, March 12-15, 2006, Basel, Switzerland. PMID- 17118787 TI - The T-cell antigen receptor: a logical response to an unknown ligand. AB - The immune system can be roughly divided into innate and adaptive compartments. The adaptive compartment includes the B and T lymphocytes, whose antigen receptors are generated by recombination of gene segments. The consequence is that the creation of self-reactive lymphocytes is unavoidable. For the host to remain viable, the immune system has evolved a strategy for removing autoimmune lymphocytes during development. This review discusses how T lymphocytes are generated, how they recognize antigens, and how their antigen receptor directs the removal of self-reactive T cells. PMID- 17118788 TI - Two-dimensional solid-state NMR applied to a chimeric potassium channel. AB - Solid-state NMR (ssNMR) represents a spectroscopic method to study membrane protein structure and dynamics in lipid bilayers. We present two-dimensional correlation experiments conducted on a fully [13C,15N] labeled version of a chimeric potassium (KcsA-Kv1.3) channel. Data obtained by using two different ion concentrations suggest a structural conservation of the selectivity filter region. SsNMR experiments conducted at two different temperatures point to differential molecular dynamics of the channel. PMID- 17118789 TI - Characterization of monoclonal antibodies directed against the rat neurotensin receptor NTS1. AB - G protein-coupled receptors (GPCRs) are integral membrane proteins that mediate cellular responses to a variety of ligands and represent major drug targets. Despite their medical importance, detailed structural information is limited because only one GPCR has been crystallized and its structure determined. To develop tools to aid in the formation of well-ordered crystals, we generated monoclonal antibodies with high affinity to the rat neurotensin receptor. All antibodies bound to the C-terminus of the receptor, which may reflect the selection strategy used to identify high-affinity binders. Further characterization revealed that some antibodies bound to the receptor in a sodium chloride sensitive manner, but others did not. Epitope mapping revealed distinct antigenic regions within the receptor C-terminus. Tight binding of Fab fragments to the receptor was verified by size exclusion chromatography. PMID- 17118790 TI - The active and the inactive form of the hAT1 receptor have an identical ligand binding environment: an MPA study on a constitutively active angiotensin II receptor mutant. AB - Several models of activation mechanisms were proposed for G protein-coupled receptors (GPCRs), yet no direct methods exist for their elucidation. The availability of constitutively active mutants has given an opportunity to study active receptor conformations within acceptable limits using models such as the angiotensin II type 1 (AT1)1 receptor mutant N111G-hAT1 which displays an important constitutive activity. Recently, by using methionine proximity assay, we showed for the hAT1 receptor that TMD III, VI, and VII form the ligand-binding pocket of the C-terminal amino acid of an antagonistic AngII analogue. In the present contribution, we investigated whether the same residues would also constitute the ligand-binding contacts in constitutively activated mutant (CAM) receptors. For this purpose, the same Met mutagenesis strategy was carried out on the N111G double mutants. Analysis of 43 receptors mutants in the N111G-hAT1 series, photolabeled and CNBr digested, showed that there were only subtle structural changes between the wt-receptor and its constitutively active form. PMID- 17118791 TI - Synthesis of an agonistic, difluoro-azido photolabel of angiotensin II and labeling of the AT1 receptor: transmembrane domains 3, 6, and 7 form the ligand binding pocket. AB - p-Azido-phenylalanine has been frequently used for photoaffinity labeling of target proteins such as the angiotensin receptors. However, chemical studies showed that simple aryl nitrenes first react intramolecularly, forming a semistable cyclic keteneimine and then reacting with nucleophile residues in the target structure like those of lysine and arginine. We synthesized 3,5-difluoro-4 azidophenylalanine where the formation of the keteneimine is prevented and where photoincorporation should be due to nonselective nitrene insertion only. This new amino acid was introduced in position 8 of angiotensin II and compared with the corresponding azidophenylalanine peptide using human AT1 receptor as target. The new photolabel maintained full agonist activity and a similar yield of photolabeling but without the previously observed gradual hydrolysis. Several selective proteolyses of the labeled receptor indicate that the new photolabel forms three simultaneous contact regions on the hAT1 receptor, suggestive of a nonselective behavior of the photolabel. A major contact was established in the sixth transmembrane domain but also in the third and seventh domain. Our results are in excellent agreement with those recently obtained from methionine proximity assay studies. PMID- 17118792 TI - A rationally designed synthetic mimic of the discontinuous CD4-binding site of HIV-1 gp120. AB - Synthetic mimetics of the CD4-binding site of HIV-1 gp120 are promising candidates for HIV-1 entry inhibition, as well as immunogen candidates for the elicitation of virus-neutralizing antibodies. On the basis of the crystal structure of gp120 in complex with CD4, we have used a recently introduced strategy for the generation of structurally diverse scaffolds to design and synthesize a scaffolded peptide, in which three fragments, making up the sequentially discontinuous binding site of gp120 for CD4, are presented in a nonlinear and discontinuous fashion through a molecular scoffold, which restrains conformational flexibility. The affinities of this molecule to CD4, as well as to the broadly neutralizing antibody mAb b12, whose epitope overlaps the CD4-binding site of gp120, were determined in competitive binding assays. PMID- 17118793 TI - Sensing, threading, orienting, and cutting polymers with rigid-rod pores. AB - This short review describes synthetic pores that are made from rigid-rod molecules and can bind oligo-and polymers such as polyacetylenes, p-oligophenyls, terpenoids, polypeptides, polysaccharides, and oligonucleotides. The spotlight is on recent breakthroughs to image the longtime elusive pore-polymer host-guest complexes as single giant pseudorotaxanes. PMID- 17118794 TI - Entirely artificial signal transduction with adrenaline. AB - Multifunctional transmembrane-building blocks with recognition sites for adrenaline on one end and the reaction partners for an SN2 reaction on the opposite end have been embedded in DPPC-liposomes. These doped vesicles can be quantitatively reduced at their disulfide head groups by externally added reducing agents; their composition and chemical processes taking place within can be monitored by NMR spectroscopy and--with limitations--by UV/Vis spectroscopy. Attempted release of thiopyridine as a second messenger into the interior of the liposome on external adrenaline addition could not be proven unambiguously because the detection system does not fulfill the necessary rigorous specificity and sensitivity requirements. PMID- 17118795 TI - Recognition of neurohormones of the NPY family by their receptors. AB - In this review a structural approach developed to answer the question whether hormones from the neuropeptide Y (NPY) family are recognized directly from solution or from the membrane-bound state is described. The chosen strategy is built onto a comparison of a set of peptides with well-known pharmacology and investigates whether similarities of structures of pharmacologically related peptides are higher in solution or in the membrane-bound state. Moreover, we have established the membrane-association mode of these peptides and contributed to our understanding of the structural features of these hormones both when placed in bulk solution and when bound to membranes. As a result we propose a receptor recognition pathway that includes initial association with the membrane and requires the peptides to come off the membrane to diffuse into the binding pocket of the receptor. This review also presents methodology recently developed by us to simulate the structural transition the peptides undergo when diffusing from bulk solution onto the membrane. PMID- 17118796 TI - Opsin oligomerization in a heterologous cell system. AB - Using bioluminescence resonance energy transfer (BRET) we studied opsin oligomerization in heterologous expression systems and quantitatively assessed its oligomerization state. BRET2 saturation and competition experiments were performed with live COS-7 cells expressing Rluc-and GFP2-tagged receptor constructs. BRET2 saturation curves obtained were hyperbolic, and the calculated oligomerization state (N = 1 for dimers) suggested that opsin (N = 1.34 +/- 0.25) forms higher oligomers. Very high BRET2 values obtained for the opsin homo-dimer pair indicated a large energy transfer efficiency (E) and for cases where E >> 0.1 a modified saturation curve was proposed. The existence of homo-dimer complexes was additionally supported by competition assay results and was also observed in HEK-293 cells. Furthermore, evidence was provided for homo-and hetero dimerization of family A (beta2-adrenergic) and B (gastric inhibitory polypeptide, GIP) receptors. In summary, these experiments demonstrate homo-and hetero-dimerization for opsin, beta 2-adrenergic, and GIP receptors. PMID- 17118797 TI - Global down-regulation of gene expression in the brain using RNA interference, with emphasis on monoamine transporters and GPCRs: implications for target characterization in psychiatric and neurological disorders. AB - RNA interference (RNAi) is a natural mechanism for regulating gene expression, which exists in plants, invertebrates, and mammals. We investigated whether non viral infusion of short interfering RNA (siRNA) by the intracerebroventricular route would enable a sequence-specific gene knockdown in the mouse brain and whether the knockdown translates into disease-relevant behavioral changes. Initially, we targeted enhanced green fluorescent protein (EGFP) in mice overexpressing EGFP. A selective knockdown of both EGFP protein and mRNA was observed throughout the brain, with lesser down-regulation in regions distal to the infusion site. We then targeted endogenous genes, encoding the dopamine (DAT) and serotonin transporters (SERT). DAT-siRNA infusion in adult mice produced a significant down-regulation of DAT mRNA and protein and elicited hyperlocomotion similar, but delayed, to that produced on infusion of GBR-12909, a potent and selective DAT inhibitor. Similarly, SERT-siRNA infusion resulted in significant knockdown of SERT mRNA and protein and elicited reduced immobility in the forced swim test similar to that obtained on infusion of citalopram, a very selective and potent SSRI. Application of this non-viral RNAi approach may accelerate target validation for neuropsychiatric disorders that involve a complex interplay of gene(s) from various brain regions. PMID- 17118798 TI - Novel GPCR screening approach: indirect identification of S1P receptor agonists in antagonist screening using a calcium assay. AB - To elucidate the physiological function of sphingosine 1-phosphate receptors 1-3 (S1P1-3) we aimed to identify selective ligands for these GPCRs. S1P2 and S1P3 are coupled to Gq, and are, therefore, linked to the phospholipase C/IP3/calcium pathway. S1P1 is solely coupled to Gi and was artificially linked to calcium signaling by coexpression of Galpha 16. The three receptors desensitized on challenge of cells with an agonist (i.e., agonists appeared as antagonists in a second calcium measurement). We screened a compound library for inhibitors of S1P stimulated calcium signals, and we could identify agonists and antagonists with a single measurement. Agonism and antagonism were confirmed by recording compound and S1P-induced calcium signals from the same assay well. For the three receptors, we found a reciprocal correlation of agonism and "apparent" antagonism of agonists. In addition, agonists indirectly discovered by this approach do not promote calcium mobilization through endogenous GPCRs. PMID- 17118799 TI - Regulation of sodium and calcium channels by signaling complexes. AB - Membrane depolarization and intracellular calcium transients generated by activation of voltage-gated sodium and calcium channels are local signals, which initiate physiological processes such as action potential conduction, synaptic transmission, and excitation-contraction coupling. Targeting of effector proteins and regulatory proteins to ion channels is an important mechanism to ensure speed, specificity, and precise regulation of signaling events in response to local stimuli. In this article, we review recent experimental results showing that sodium and calcium channels form local signaling complexes, in which effector proteins, anchoring proteins, and regulatory proteins interact directly with ion channels. The intracellular domains of these channels serve as signaling platforms, mediating their participation in intracellular signaling processes. These protein-protein interactions are important for efficient synaptic transmission and for regulation of ion channels by neurotransmitters and intracellular second messengers. These localized signaling complexes are essential for normal function and regulation of electrical excitability, synaptic transmission, and excitation-contraction coupling. PMID- 17118800 TI - Receptors for protons or lipid messengers or both? AB - The subfamily of G protein-coupled receptors comprising GPR4, OGR1, TDAG8, and G2A was originally characterized as a group of proteins mediating biological responses to the lipid messengers sphingosylphosphorylcholine (SPC), lysophosphatidylcholine (LPC), and psychosine. We challenged this view by reporting that OGR1 and GPR4 sense acidic pH and that this process is not affected by concentrations of SPC or LPC previously reported as agonistic. The original publications describing GPR4, OGR1, and G2A as receptors for LPC or SPC have now been retracted, and the first studies exploring receptors of this family as pH sensors in physiology have appeared. Here we review the status of this field and we confirm that GPR4, OGR1, and TDAG8 should be considered as proton sensing receptors. Negative regulation of these receptors by high micromolar concentrations of lipids appears not specific in our experiments. PMID- 17118801 TI - Computational studies of ligand-receptor interactions in bitter taste receptors. AB - Phenylthiocarbamide tastes intensely bitter to some individuals, but others find it completely tasteless. Recently, it was suggested that phenylthiocarbamide elicits bitter taste by interacting with a human G protein-coupled receptor (hTAS2R38) encoded by the PTC gene. The phenylthiocarbamide nontaster trait was linked to three single nucleotide polymorphisms occurring in the PTC gene. Using the crystal structure of bovine rhodopsin as template, we generated the 3D structure of hTAS2R38 bitter taste receptor. We were able to map on the receptor structure the amino acids affected by the genetic polymorphisms and to propose molecular functions for two of them that explained the emergence of the nontaster trait. We used molecular docking simulations to find that phenylthiocarbamide exhibited a higher affinity for the target receptor than the structurally similar molecule 6-n-propylthiouracil, in line with recent experimental studies. A 3D model was constructed for the hTAS2R16 bitter taste receptor as well, by applying the same protocol. We found that the recently published experimental ligand binding affinity data for this receptor correlated well with the binding scores obtained from our molecular docking calculations. PMID- 17118802 TI - Regulation of g protein-coupled receptor signaling by a-kinase anchoring proteins. AB - Specificity of transduction events is controlled at the molecular level by scaffold, anchoring, and adaptor proteins, which position signaling enzymes at proper subcellular localization. This allows their efficient catalytic activation and accurate substrate selection. A-kinase anchoring proteins (AKAPs) are group of functionally related proteins that compartmentalize the cAMP-dependent protein kinase (PKA) and other signaling enyzmes at precise subcellular sites in close proximity to their physiological substrate(s) and favor specific phosphorylation events. Recent evidence suggests that AKAP transduction complexes play a key role in regulating G protein-coupled receptor (GPCR) signaling. Regulation can occur at multiple levels because AKAPs have been shown both to directly modulate GPCR function and to act as downstream effectors of GPCR signaling. In this minireview, we focus on the molecular mechanisms through which AKAP-signaling complexes modulate GPCR transduction cascades. PMID- 17118803 TI - Localization of novel adiponectin receptor constructs. AB - Adiponectin is one of the most abundant fat-derived hormones involved in a multitude of metabolism pathways. The receptors AdipoR1 and AdipoR2 of this metabolically active protein have been identified recently. AdipoR1 and AdipoR2 are most abundantly expressed in the skeletal muscle and in the liver, respectively. It has been postulated that although they both consist of seven transmembrane helices, they are distinct from other G protein-coupled receptors (GPCRs). We cloned both receptors as fusion proteins with enhanced yellow fluorescent protein (YFP) to determine their localization and orientation in the cell membrane. By confocal microscopy and immune staining we demonstrated that both receptor-YFP-fusion proteins are integral membrane proteins with the predicted topology--an intracellular N-terminus and an extracellular C-terminus. In parallel, comparative experiments were performed with the NPY Y2-receptor, a classical rhodopsin-like GPCR. PMID- 17118804 TI - Effect of dominant-negative epidermal growth factor receptors on cardiomyocyte hypertrophy. AB - Angiotensin II (AngII) induces heart growth via cardiomyocyte hypertrophy, and central to this is the capacity of the type 1 AngII receptor (AT1R) to "transactivate" epidermal growth factor receptors (EGFRs)--a family with four main subtypes (HER1-4)--although the exact molecular mechanism remains unresolved. In this study, the pharmacological inhibition of AngII-stimulated ERK1/2 activation and cardiomyocyte hypertrophy by increasing concentrations of an EGFR inhibitor, AG1478, indicated that other EGFR subtypes, in addition to HER1, may be involved. We constructed expression vectors and adenoviruses expressing truncated mutant versions of HER1, HER2, and HER4 and determined their capacity to act as dominant-negative inhibitors when co-transfected with full length EGFRs. It is surprising that adenoviral-mediated expression of these truncated EGFRs in cardiomyocytes led to paradoxical, ligand-independent increases in cardiomyocyte hypertrophy and unusual morphological changes. These results challenge our perception of AT1R-mediated EGFR transactivation and imply that truncated EGFRs may affect cell function through unconventional mechanisms. PMID- 17118805 TI - Control of peroxisome proliferator-activated receptor fate by the ubiquitinproteasome system. AB - Peroxisome proliferator-activated receptor (PPAR) alpha, gamma, and delta belong to the nuclear hormone receptor superfamily of ligand-activated transcription factors. PPARs regulate metabolic, developmental, and differentiation pathways and play important roles in human diseases, such as diabetes, atherosclerosis, cancer, and chronic inflammation. PPARs are the targets of drugs of widespread clinical use and represent promising targets for discovery of new therapeutics. The interaction of PPARs with the ubiquitin-proteasome system (UPS) has been the subject of limited investigation. The UPS plays an important role in regulating the levels and modulating ligand-dependent and-independent activity of nuclear receptors. This review highlights the current knowledge regarding the interactions of the UPS with PPARs and focuses on the differential regulation of the level and activity of the PPAR isotypes by the UPS in response to selective ligands. Understanding the connections between the UPS and PPARs can provide insights in the actions of existing drugs and raise the possibilities for development of more effective PPAR-based therapeutics. PMID- 17118806 TI - Comparative binding of 125I-and 99mTc-d-labeled native and glycated low-density lipoprotein to human microvascular endothelial cells-potential for atherosclerosis imaging? AB - Native (n), glycated (g), and glycoxidated (go) low-density lipoproteins (LDL) were labeled with 125I or 99mTc, and the labeling efficiency and binding were assessed for potential use of these LDL compounds in imaging analysis of atherosclerotic lesions (PPAR-gamma receptors) by determining the number of specific receptors for nLDL, gLDL or goLDL on human microvascular endothelial cells as well as the KDs using either 125I-or 99mTc-labeled LDLs. The specific activity of labeled gLDL and goLDL was much higher (for goLDL 20 times higher) than that of nLDL. Gel filtration of labeled LDLs revealed, however, that 99mTc g/goLDL is significantly degraded by the labeling reaction. No fragmentation was observed for 99mTc-nLDL and all the 125I-labeled LDL forms. Binding studies using both 125I-and 99mTc-nLDL indicated a weak binding affinity (KD 10- 7mol/L) to human microvascular endothelial cells. The binding affinity of 125I-g/goLDL to these cells was significantly higher (KD 10- 9mol/L) and could be increased further by preactivation of the endothelial cells using TNFalpha. Incubation with 99mTc-goLDL, however, did not result in specific binding of the ligand, possibly as a consequence of the fragmentation of the lipoprotein during the labeling. Scatchard transformation of the binding data with 99mTc-gLDL revealed the presence of only a few binding sites. This was in contrast to the results obtained with 125I-labeled gLDL, which revealed a much higher membrane density of scavenger receptors for this ligand. We conclude that for in vitro binding studies as well as for potential in vivo imaging, only 125I-labeled goLDL should be used, whereas nLDL may be applied as 125I-or 99mTc-labeled ligand. PMID- 17118807 TI - New natural noncannabinoid ligands for cannabinoid type-2 (CB2) receptors. AB - Since the discovery that Delta 9-tetrahydrocannabinol and related cannabinoids from Cannabis sativa L. act on specific physiological receptors in the human body and the subsequent elucidation of the mammalian endogenous cannabinoid system, no other natural product class has been reported to mimic the effects of cannabinoids. We recently found that N-alkyl amides from purple coneflower (Echinacea spp.) constitute a new class of cannabinomimetics, which specifically engage and activate the cannabinoid type-2 (CB2) receptors. Cannabinoid type-1 (CB1) and CB2 receptors belong to the family of G protein-coupled receptors and are the primary targets of the endogenous cannabinoids N-arachidonoyl ethanolamine and 2-arachidonoyl glyerol. CB2 receptors are believed to play an important role in distinct pathophysiological processes, including metabolic dysregulation, inflammation, pain, and bone loss. CB2 receptors have, therefore, become of interest as new targets in drug discovery. This review focuses on N alkyl amide secondary metabolites from plants and underscores that this group of compounds may provide novel lead structures for the development of CB2-directed drugs. PMID- 17118808 TI - GABAA receptors in central nervous system disease: anxiety, epilepsy, and insomnia. AB - Brain function is based on an exquisite balance between excitatory and inhibitory neurotransmission. GABAergic neurons provide the major inhibitory control. By controlling spike timing and sculpting neuronal rhythms they play a key role in regulating behavior. GABAergic neurons are highly diverse and operate with a corresponding diversity of GABAA receptor subtypes. In this article, the contribution of GABAA receptor deficits to central nervous system disorders, in particular anxiety disorders, epilepsy, schizophrenia and insomnia, is reviewed. PMID- 17118809 TI - Predictors of smoking cessation processes among secondary school students. AB - Many adolescents want to quit and have specific ideas on how they want to go about it. This study extended the search for factors related to different aspects of quitting. Four cross-sectional surveys of Queensland (Australia) secondary schools every three years over the past decade (1993, 1996, 1999, and 2002). A total of 9993 school students in grades 8 to 12 completed the survey. The data for 2451 students who had smoked in the last week (51% female) were used for the analysis of smoking cessation outcomes. The outcome measures for the study represented various smoking cessation outcomes, such as a desire to quit, attempts to quit, quitting for a week or more, and an intention not to be smoking in the following year. More than 60% of adolescent smokers are involved in the process of smoking cessation. The strongest predictor of cessation was whether or not students had actively influenced other students not to smoke. This finding supports the concept of involving adolescents more actively in prompting their peers to try quitting and supporting their efforts to quit smoking. PMID- 17118810 TI - Effects of craving self-report measurement on desire for heroin in opioid dependent individuals. AB - This study aims to investigate whether the administration of questionnaires measuring subjective craving induces more craving for opiates compared to questionnaires measuring other subjective states. The study was conducted in 2000. The sample was composed of 53 patients that were treated as inpatients and outpatients for their opioid dependence syndrome. Participants were assigned randomly into four groups. Both a craving and negative affect condition were presented to each group. The administration of the Visual Analogue Scale (VAS) preceded and followed each condition. Findings of the present study show that the administration of questionnaires measuring instant as well as general craving does not have a distinctive effect on measures of unidimensional craving compared to a questionnaire focused on anxiety and depression states. Because of the small sample, the results should be interpreted cautiously. PMID- 17118811 TI - Anabolic steroid use in high school students. AB - A total of 1351 high school students (52.3% males, 47.7% females) with mean age 17.5 years (SD = 2.2) from randomized school classes in Hordaland County, Norway, participated in an Internet survey conducted in 2004 about the lifetime use of anabolic steroids and personal acquaintance with at least one user of anabolic steroids. In addition to questions about anabolic steroids the participants completed the Hospital Anxiety and Depression Scale and the Alcohol Use Disorders Identification Test. They also answered questions about demography, smoking, and narcotic use. The lifetime prevalence for use of anabolic steroids was 3.6% for males and 0.6% for females. In all, 27.9% of the respondents reported having at least one acquaintance that used or had used anabolic steroids. Use of anabolic steroids and having acquaintances using such drugs were strongly related to use of other drugs such as alcohol, nicotine, and narcotics. Implications for prevention are discussed and the study's limitations are noted. PMID- 17118812 TI - Focus on secondary prevention: implications of a study on intervention in social networks. AB - Screening and brief intervention (SBI) as a method within secondary prevention of alcohol use-related problems has experienced enormous attention and interest over the past 20 years. Initial results were very promising and great effort was put into designing screening instruments and developing different kinds of interventions. However, at the same time, the approach has generated problems in its own right. Some of these problems relate to standardization of instruments and some to criteria of inclusion into samples; others relate to the focus on the individual drinking style independent from social influences. In light of these problems, it is necessary to elaborate on the theoretical foundation, as well as on the methods used in SBI. This article introduces a method for intervening in social networks with important implications for SBI. By putting emphasis on the motivational part of SBI and including social networks in the field of intervention, it may be assumed that the approach will produce better results than heretofore. The results from a Danish qualitative study on intervening into the social network of social drinkers are presented. This study was carried out between 1991 and 1999 and involved 13 adult Danes between 35 and 45 years of age. They all had a weekly alcohol consumption of 120 to 360 g of pure alcohol and they all considered themselves to be social drinkers. The results of this study show that enhanced awareness of the person's own drinking, as well as that of peers, may influence decisions about specific drinking situations. These findings and the perspectives for SBI are discussed. Recommendations for further research are also presented. PMID- 17118813 TI - What Chinese adolescents think about quitting smoking: a qualitative study. AB - The aim of this study was to investigate the attitudes of Chinese adolescents toward smoking, giving up smoking, and smoking cessation programs presently available. The study was a qualitative study carried out in 2002 by focus groups of 32 male secondary school students in Hong Kong who were either current smokers or had recently given up smoking. Subjects were students (grades 8-10) attending two full-day secondary schools in Hong Kong. Participants did not feel the need to make any serious psychological preparation for quitting. They underestimated the addictive nature of cigarette smoking and felt that they could choose to quit smoking at any time with little difficulty. Several barriers to quitting were reported, including boredom, peer influence, the urge to smoke, school work pressure, the wish to do something with their hands, difficulty in concentrating, and the ready availability of free cigarettes from peers. Those who had attempted to quit smoking (26/32) reported that peer influence and boredom were the main reasons why they started smoking and insisted that willpower and determination could have helped them in their quitting attempt. Participants were unanimous that pressure or encouragement from teachers, parents, or girlfriends did not help them to stay off cigarettes. Most (24/32) of the current smokers knew that smoking cessation services were available in Hong Kong, only 50% (12/24) of those who knew had made use of such services. None of the participants were able to identify any effective way of quitting smoking, though some suggested that the best practical measure was to avoid friends who smoked. The study suggests that attempts to persuade young people to quit smoking might benefit if they were framed to address issues such as the strong influence of their peers, the ease with which tobacco products can be obtained, the casual attitudes of young people toward smoking cessation, the perceived pros and cons of quitting, and (given that underage smoking is frowned upon by many parents and teachers) the need to respect confidentiality when offering support. PMID- 17118814 TI - Integrated HIV care: HIV risk outcomes of pregnant substance abusers. AB - Identifying contextual factors that may influence the effects of HIV risk reduction strategies aimed at inner-city, minority female populations may be critical to interrupting the alarming trends in seroprevalence in the United States, especially among pregnant women with substance use-related problems. The objective of this Phase I, NIDA-funded project was to determine which contextual and cognitive factors were most predictive of HIV outcomes in this population. Eighty-one HIV-negative women were enrolled in a maternal addiction program with a cognitive-behavioral HIV risk-reduction component. Measures were administered between 1996 and 1998, one week post-admission, and follow-up assessments were conducted on 69 participants at 6 months post-discharge Paired t-tests were conducted to assess changes in sex risk behaviors, HIV/AIDS-related knowledge, and HIV risk-related attitudes/behaviors. Hierarchical regression analyses were conducted to investigate the relationship between attitudes/knowledge with contextual variables. At six months follow-up, there were significant increases in favorable condom attitudes (t = 3.36, p = .01) and in factual knowledge regarding HIV (t = 3.20, p = .01), with a significant decrease in the number of sexual partners (t = 2.21, p = .05). Hierarchical regression analysis revealed that the strongest predictors of the number of partners--a key outcome variable- were alcohol use, intentions to engage in safer sex behaviors, psychiatric symptoms, and a history of physical abuse (F[11, 57] = 6.58, p < .001). This study also reinforces the strategic importance of utilizing substance user treatment programs as crucial vehicles for integrating HIV risk-reduction strategies. Additionally, it will further guide the design of effective procedures to test the feasibility of an integrated HIV risk-reduction intervention for a larger randomized controlled study. The study's limitations are noted. PMID- 17118815 TI - Alcohol use severity and HIV sexual risk among juvenile offenders. AB - Guided by the Information Motivation Behavioral Skills (IMB) model, we examine the alcohol severity/sexual risk relationship for juvenile offenders who are at extreme risk for HIV/AIDS due to situational vulnerabilities, substance abuse,1 and personality factors. Sexual risk behavior was analyzed by levels of alcohol use among 634 ethnically diverse adolescents in Miami between 1998 and 2002. Adolescents with the highest levels of alcohol use reported significantly higher levels of total and unprotected sexual activity and sex acts proximate to drinking. Alcohol use related problems require more attention by HIV interventionists. Alcohol severity may reduce the effectiveness of HIV interventions that do not address concurrent substance use. PMID- 17118816 TI - Buddhism and adolescent alcohol use in Thailand. AB - A sample of 2019 Thai secondary school students in grades equivalent to U.S. 10 through 12 completed a 43-item alcohol expectancy questionnaire in June 2000. Factor analysis revealed four factors: (a) positive expectancies, (b) negative expectancies, (c) sex and power expectancies, and (d) religious expectancies. Practicing Buddhists were less likely to drink than nonpracticing Buddhists and had fewer positive and more negative expectancies about alcohol. Among students who did drink, Buddhist beliefs did not appear to influence whether or not they were binge drinkers. Buddhist beliefs may influence decisions to drink but not decisions related to drinking patterns. PMID- 17118817 TI - Non drug use- and drug use-specific spirituality as one-year predictors of drug use among high-risk youth. AB - The present article explored two different dimensions of spirituality that might tap negative and positive relations with adolescent drug use over a 1-year period. Non-drug-use-specific spirituality measured how spiritual the person believes he or she is, participation in spiritual groups, and engagement in spiritual practices such as prayer, whereas drug-use-specific spirituality measured using drugs as a spiritual practice. Self-report questionnaire data were collected during 1997-1999 from a sample of 501 adolescents in 18 continuation high schools across southern California. Participants ranged in age from 14 to 19 and were 57% male, with an ethnic distribution of 34% White, 49% Latino, 5% African American, 7% Asian, and 5% other. A series of general linear model analyses were conducted to identify whether or not two different spirituality variables predict drug use (cigarettes, alcohol, marijuana, hallucinogens, and stimulants) at 1-year follow-up. After controlling for baseline drug use, non drug-use-specific spirituality was negatively predictive of alcohol, marijuana, and stimulant use, whereas drug-use-specific spirituality failed to be found predictive of these variables one year later. Conversely, drug-use-specific spirituality was positively predictive of cigarette smoking and hallucinogen use, whereas non-drug-use spirituality failed to be found predictive of these variables. Our results provide new evidence that suggests that spirituality may have an effect on drug use among adolescents. The drug-use-specific measure of spirituality showed "risk effects" on drug use, whereas the other measure resulted in "protective effects," as found in previous research. Knowledge of the risk and protective patterns and mechanisms of spirituality may be translated into future drug use prevention intervention programs. PMID- 17118818 TI - Patterns of drug use among drug misusers in Sweden. Gender differences. AB - The goal of the article is to provide information about polydrug abuse and drug misuse patterns in Sweden among women and men. The data has been taken from a 1998 national survey of "Heavy"/severe drug misuse in Sweden, project "MAX-98" (Olsson, Adamsson-Wahren, & Byqvist, 2001). The drug misusers were reported by various government agencies, including health services, social services, police, and correctional treatment facilities on a special form. One of the significant gender differences that emerged was that a greater percent of the women in the survey used and injected amphetamines, injected opiates, and used tranquilizers/sedatives, while a greater percent of the men smoked cannabis, smoked heroin, and misused alcohol. Furthermore, the most common combinations for both genders was amphetamines + cannabis, followed by amphetamines + heroin + cannabis. Alcohol played a large role for the narcotics users. Heroin as a primary drug has grown in the age groups under 35. The trends document that the use of ecstasy as well as chemical CNS-stimulating/hallucinogenic drugs has grown, that polydrug use has increased compared with earlier surveys, and that the methods of ingestion have changed. It is therefore more precise today to speak of different types of polydrug users than about users of exclusively one drug. PMID- 17118820 TI - Percutaneous Coronary Intervention (PCI): "The Ugly Duckling?". PMID- 17118821 TI - Are the symptoms of myocardial infarction different in men and women, if so, will there be any consequences? PMID- 17118822 TI - Effect of postconditioning on coronary blood flow velocity and endothelial function in patients with acute myocardial infarction. AB - OBJECTIVES: To observe the effect of postconditioning on coronary blood flow velocity and endothelial function in acute myocardial infarction (AMI). METHODS: We recruited patients with AMI who underwent primary PCI. The patients were randomized to an ischemic-reperfusion group (IR group) and a postconditioning group. Corrected TIMI frame count (CTFC), Wall motion score index (WMSI), Creatine phosphokinase (CK), CK-MB, malondialdehyde (MDA) were measured. Arterial endothelial function was studied non-invasively by echo Doppler technique. RESULTS: Patients with postconditioning had much faster CTFC. After 8 weeks, the WMSI in Postcond group was significantly larger. There was a significant negative correlation between WMSI and CTFC in IR group and Postcond group(r = - 0.90, p < 0.01; r = - 0.79, p < 0.01). The peaks of CK and CK-MB of Postcond group were much lower than those of the IR group, and MDA-reactive products were significantly lower than those of the IR group after PCI. The endothelium dependent vasodilatory function was improved in the Postcond group. CONCLUSION: Postconditioning is a simple interventional procedure for salvage of the coronary endothelial function and increasing coronary blood flow velocity. PMID- 17118823 TI - Postconditioning in rats and mice. AB - OBJECTIVE: For subsequent studies on the molecular mechanisms of postconditioning, we aimed to identify a robust postconditioning protocol in rat and mouse heart. DESIGN: Isolated rat hearts were subjected to different postconditioning protocols (study 1 and 2). The protection was compared to preconditioning. Rats (study 3) in vivo in two different laboratories were postconditioned. Isolated mouse hearts (study 4) and mice in vivo (study 5) were postconditioned. RESULTS: Postconditioning did not protect isolated, perfused rat hearts, however, preconditioning improved function and reduced infarct size. Postconditioning tended to protect rat hearts in vivo in one laboratory (p = 0.10), whereas protection was seen in the other laboratory (infarct size 51+/-11% vs controls 62+/-3%, p = 0.01). Postconditioned mouse hearts were protected, both ex vivo (16+/-9% vs controls 33+/-18%, p = 0.02) and in vivo (21+/-5% vs 42+/-7%, p < 0.001). CONCLUSIONS: Rat hearts are less suitable for studies of mechanisms of postconditioning. The results suggest that the signaling pathways differ between pre- and postconditioning. Mouse hearts were strongly protected by postconditioning, and genetically engineered mice may be useful for postconditioning research. PMID- 17118824 TI - Women with myocardial infarction are less likely than men to experience chest symptoms. AB - OBJECTIVES: To assess similarities and differences between women and men regarding experience and interpretation of symptoms of acute myocardial infarction before hospital admission. DESIGN: 149 women and 384 men responded to a questionnaire two weeks after hospital discharge. RESULTS: Men were more likely than women to report chest symptoms. Women were more likely to complain of nausea, palpitations, dyspnoea, fainting, pain in the back and pain between the scapulas than men. Irrespective of sex only half the patients interpreted their symptoms as cardiac. Early onset of chest symptoms, previously diagnosed angina, symptoms in accordance with expectations, symptoms stronger than expected and familiarity with symptoms, all contributed to a cardiac attribution among both sexes. A diagnosis of hypercholesterolemia and prescribed nitroglycerin was related to a cardiac attribution among men only. CONCLUSION: Women were less likely to experience chest symptoms and more likely to experience atypical symptoms than men. Symptom experience and the patients' expectations of symptoms influenced interpretation and attribution among both sexes. PMID- 17118825 TI - The Swedish Heart Surgery Register: data quality for proximal thoracic aortic operations. AB - OBJECTIVES: To review the data quality and validity in the nationwide Swedish Heart Surgery register for patients operated on the proximal thoracic aorta. DESIGN: Medical records from a random sample of 300 patients in The Swedish Heart Surgery register were reviewed with register data items systematically re reported. Variable reporting frequency, proportion of adequately reported data, and number and correctness of diagnostic and procedural codes were analysed. RESULTS: After exclusions, 251 patients (84%) remained in the analysis. Reporting frequency for individual items varied from 12% to 100% (median 61%). For core variables, reporting frequency was 96%-100%. In 40 of 43 (93%) reviewed variables, registry data were at least 85% correct. A total of 485 diagnoses and 673 procedures were reported, compared to 617 diagnoses and 758 procedures identified in the review process. CONCLUSIONS: The register data quality and validity for patients operated on the proximal thoracic aorta was satisfactory overall, but need further improvement for complications. The register coverage and completeness was very high. Register-based reports should be accompanied by review of data quality. PMID- 17118826 TI - Scintigraphic demonstration of myocardial perfusion and ischaemia associated with coronary artery bypass grafting. AB - OBJECTIVES: To assess if myocardial perfusion scintigraphy (MPS) at rest can be of value in elucidating myocardial perfusion, ischaemia and perioperative myocardial infarction (PMI) associated with coronary artery bypass graft (CABG) surgery. DESIGN: This was a prospective randomized study of patients undergoing elective CABG. Forty-eight patients in the control group underwent serial ECG recordings and measurements of CK-MB and cTnT. Fifty-four patients in the study group were additionally examined with MPS preoperatively and 2-4 days and 6 weeks postoperatively. RESULTS: The study showed a highly significant (p < 0.001) improvement in myocardial radionuclide uptake from preoperatively to 2-4 days postoperatively. Judged from ECG and enzymatic changes, two control patients and one study patient only had PMI and no additional cases of PMI were demonstrated by MPS. CONCLUSION: MPS at rest showed that CABG significantly improved myocardial perfusion, by demonstrating an increase in radionuclide uptake. In diagnosing PMI, we found that MPS provided no additional information beyond cardiac biochemical markers and ECG changes. PMID- 17118827 TI - Changes of plasma asymmetric dimethylarginine levels after coronary artery bypass grafting. AB - OBJECTIVES: We investigated whether coronary artery bypass grafting affects plasma asymmetric dimethylarginine (ADMA) concentrations and whether precardioplegic hyperoxia influences ADMA release from the heart. DESIGN: Twenty two patients were randomized into control (n = 11) and hyperoxia (n = 11, ventilated with >96% oxygen before cardiopulmonary bypass) groups. Arterial and coronary sinus blood was sampled before cardioplegia and during early reperfusion. Arterial samples were drawn 60 min after declamping of the aorta, and on the first postoperative day. RESULTS: Baseline arterial values of ADMA were not different between groups (0.59+/-0.18 micro mol/l control, 0.63+/-0.13 micro mol/l hyperoxia group). Negligible release of ADMA into coronary sinus was detected 20 min after cardioplegia. A significant decrease of arterial ADMA was observed by the first postoperative morning (0.42+/-0.16 micro mol/l in control, and 0.38+/-0.07 in hyperoxia group, p < 0.01 compared to baseline). CONCLUSIONS: CABG with cardioplegia is associated with decrease of ADMA by the first postoperative morning. Reperfusion of cardioplegic heart did not result in significant release of ADMA. Pretreatment with hyperoxia had no influence on myocardial release and arterial levels of ADMA. PMID- 17118828 TI - Myocardial cooling with ice-slush provides no cardioprotective effects in aortic valve replacement. AB - OBJECTIVES: Topical cooling of the heart with ice-slush has been widely used for myocardial protection. No prospective, randomized study has evaluated the effect of ice-slush on acknowledged markers (CK-MB, troponin-T) of myocardial damage during aortic valve replacement (AVR). This was the first aim of the present study. A second aim was to examine whether performing a study per se reduced myocardial damage. DESIGN: Sixty patients undergoing AVR were receiving cold crystalloid antegrade cardioplegia every 20 min. Thirty patients were randomized to achieve additional topical cooling with ice-slush. CK-MB and troponin-T were compared between groups as well as to a group of patients undergoing AVR immediately prior to the study. RESULTS: There were no significant differences in myocardial markers between patients with or without ice-slush. However, we found significantly higher levels of troponin-T and CK-MB in patients undergoing AVR prior to start of the study. CONCLUSIONS: Topical cooling with ice-slush does not provide additional cardioprotective effects. Comparison with an historical cohort indicates that administration of crystalloid cardioplegia following a rigid protocol might reduce myocardial damage. PMID- 17118829 TI - Quantitative evaluation of a novel sensor for measuring muscular tissue flow and gases. AB - Information about muscular tissue flow is important for diagnosing, treating and monitoring patients with tissue ischemia. For this purpose an objective method which is reproducible and continuous is needed. In co-operation with the company Unisense A/S, we have developed a sensor for instantaneous and continuous monitoring of muscle tissue flow in vivo. The method is based on a flexible microsensor which emits and measures minute amounts of inert tracer gases. The objective was to evaluate the capability of the microsensor to measure varying degrees of tissue flow changes and compare these measurements with the 133Xenon Washout Technique for measuring local perfusion rates in skeletal muscle. The Unisense microsensor was tested in the gracilis muscle of six anaesthetized pigs subjected to varying degrees of muscle ischemia. We found that both tissue flow and pO2 declined in synchrony with reduced blood flow to the lower extremities. All the data from the Unisense microsensor show the same trend as the Xenon data and thus confirms that the microsensor can measure changes in tissue flow. PMID- 17118830 TI - Does preemptive stellate ganglion blockage increase the patency of radiocephalic arteriovenous fistula? AB - OBJECTIVE: Radio-cephalic arteriovenous fistulas (AVFs) have high early failure ratio. Increased sympathetic activity and spasm of radial artery during the surgery may responsible for early occlusion rate. DESIGN: Fifty patients were randomized to two groups (each containing 25 patients). Stellate Ganglion Blockade (SGB) was performed in Group 1. Another group was considered as control group (Group 2) to make statistical comparisons. All AVFs were performed under local anesthesia in both groups. RESULTS: Average fistula flow was 201.4+/-40.4 ml/min in Group 1 and 155.6+/-27.4 ml/min in Group 2 (p < 0.001). While average peak velocity of radial artery was 167.1+/-31.3 cm/sec in Group 1, it was 107.8+/ 15.8 cm/sec in Group 2 (p < 0.001). Thrill was found in all Group 1 patients, but there was thrill only 13 of the Group 2 patients (p < 0.001). Mean maturation time was 41.4+/-6.8 days after surgery in Group 1 and 77.1+/-10.5 days in Group 2 (p < 0.001). Adequate vascular access was obtained 19 patients in Group 1 and 12 patients in Group 2 (p = 0.041). CONCLUSION: AVF occlusion rate is much more common in early postoperative period. Diminished sympathetic tonus by preemptive SGB not only increases early patency rate but also increases fistula maturation rate. PMID- 17118832 TI - Rectal cancer irradiation. Long course, short course or something else? PMID- 17118833 TI - Acta Oncologica Lecture. Present status of the use of recombinant human TSH in thyroid cancer management. PMID- 17118834 TI - Radioactive iodine in thyroid medicine--how it started in Sweden and some of today's challenges. AB - In Sweden, radioactive iodine for thyroid diagnostics and therapy was introduced by Jan Waldenstrom (1906-1996) and Bengt Skanse (1918-1963). The paper describes the start of the clinical use of radioiodine, the various iodine isotopes available, measurement techniques and dosimetry. There are still problems to solve in relation to an optimal clinical use of radioiodine. One of the remaining challenges is to get consensus about the goal of the treatment of hyperthyreosis, as well as about a method for individual absorbed dose calculations. Careful dose estimates will prevent unnecessary radiation exposure and constitute a base for a future optimised radioiodine therapy. For the dose calculation, it is important to understand if there is any clinically significant temporary reduction in the ability of thyroid tissue to trap or retain 131I-iodide following prior administration of a diagnostic activity of 131I-iodide (stunning of the thyroid). This may be of special concern in connection with treatment of thyroid cancer and its metastases. Finally, the production capacity, availability and delivery of 123I have to be improved to increase clinical access to this radionuclide, which is optimal for diagnostic imaging and which gives lower absorbed dose and therefore also less risk for thyroid stunning than 131I. PMID- 17118835 TI - Thyroid cancer after exposure to radioactive 131I. AB - The thyroid gland is susceptible to radiation carcinogenesis, and the thyroid cancer risk decreases with increasing age at exposure, with a low risk above 20 years of age at exposure. The risk is best described be a linear dose-response relationship down to 0.1 Gy. Epidemiological studies of patients have not observed any increased risk for thyroid cancer after 131I exposure, but the statistical power to detect risks in children is limited. The Chernobyl accident led to substantial 131I exposure in Belarus, the Russian Federation and Ukraine. About 4000 cases of thyroid cancer have been diagnosed among those who were children and adolescents in 1986, including about 3000 in the age group 0-14 years. The risk per Gy from 131I in young subjects may be less than that seen after external low-LET radiation. A recent case-control study found a threefold risk for thyroid cancer among children from severely iodine-deficient areas, as compared with those living in lesser iodine-deficient areas. A threefold risk reduction was observed among those children receiving stable iodine compared with those not receiving iodine. PMID- 17118836 TI - Radioiodine treatment for malignant thyroid disease. AB - Radioiodine treatment for thyroid disease has been given for half a decade in Sweden. The most common indication for treatment is hyperthyroidism, when iodine uptake is high. The situation in which radioiodine treatment is used in thyroid cancer is less favourable and measures therefore have to be taken to optimize the treatment. Treatment should be performed early in the course of the disease to achieve the highest possible differentiation. Before treatment the iodine and goitrogen intake should be kept low. Stimulation of the thyrocytes by thyroid stimulating hormone (TSH) should be high. It is conventionally achieved by thyroid hormone withdrawal rendering the patient hypothyroid, or by the recently available recombinant human TSH (rhTSH) which can be recommended for ablation of the thyroid remnant after thyroidectomy and for treatment of metastases in fragile patients unable to undergo hypothyroidism. Finally, stunning--the negative effect of a prior test dose from radioactive iodine--should be avoided. PMID- 17118837 TI - Endocrine ophthalmopathy and radioiodine therapy. AB - Endocrine ophthalmopathy is to some degree present in most patients with Graves' disease. In few cases, a severe form of the condition develops and in the majority of these cases, the course of the eye problems has been influenced by the treatment for thyrotoxicosis. In this regard, radioiodine therapy has been increasingly recognized as carrying a special risk. Here, the current understanding of endocrine ophthalmopathy and the risks associated with the development of severe eye disease are discussed. The results of a retrospective investigation of patients with severe eye disease in our hospital, and the experience with corticosteroid administration following radioiodine in order to reduce the risk of ophthalmopathy, are also presented. PMID- 17118838 TI - Radioiodine therapy in non-toxic multinodular goitre. The possibility of effect amplification with recombinant human TSH (rhTSH). AB - There is no consensus regarding the optimum treatment of benign non-toxic goitre. L-thyroxine suppressive therapy is widely used, but there is poor evidence of its efficacy, and it may have serious adverse effects on health. Surgery is first choice in large goitres or if malignancy is suspected. 131I therapy results in a one-year goitre reduction of around 40% in multinodular goitres, usually with a high degree of patient satisfaction and improvement of the inspiratory capacity. The effect is attenuated with increasing goitre size. The risk of hypothyroidism is 22-58% within 5-8 years. A sufficient thyroid 131I uptake is mandatory for 131I therapy to be feasible and pre-stimulation with recombinant human TSH (rhTSH) increases this considerably. This leads to an increased absorbed thyroid dose by approx.75%, mainly in those patients with the lowest thyroid 131I uptake, and a more homogeneous intrathyroidal isotope distribution. Pre-stimulation with even a small dose of rhTSH seems to allow a reduction of the 131I activity while still achieving a mean goitre reduction of approximately 40% within a year. A significantly lower extrathyroidal radiation is achieved by this approach. With an unchanged 131I activity, rhTSH pre-stimulation improves the goitre reduction by 30-50%. However, this is at the expense of a higher rate of hypothyroidism, cervical pain and transient thyrotoxicosis. Of particular concern is the observation made in healthy persons, that rhTSH results in a transient average thyroid volume increase of 35%. A similar goitre swelling may cause problems in susceptible patients during rhTSH-augmented 131I therapy. Thus, this concept still needs a closer evaluation before routine use. PMID- 17118839 TI - Risk of parathyroid adenomas in patients with thyrotoxicosis exposed to radioactive iodine. AB - External ionizing radiation is a risk factor for primary hyperparathyroidism. Whether exposure to radioactive iodine contributes to the risk of primary hyperparathyroidism is unknown. Patients with thyrotoxicosis are often treated with radioactive iodine and its accumulation in the thyroid gland exposes the adjacent parathyroid glands to radioactivity. Six thousand and eighty two patients with thyrotoxicosis (ICD-9 = 242) were identified from medical records. In a randomly selected subcohort we assessed the frequency of treatment with radioactive iodine to be 86%. The number of patient-years at risk was 77,118. Patients with parathyroid adenomas (ICD-9 = 195.1) were recruited from the Swedish Cancer Registry. Eleven patients with parathyroid adenomas following the diagnosis of thyrotoxicosis were identified. The standard incidence ratio (SIR) compared to the reference population of approximately 900,000 was 1.14 (95% CI 0.57-2.03). The median age at exposure was 59 years and the latency period between diagnosis of thyrotoxicosis and parathyroid adenoma was 7.4 years (range <1-19 years). This study does not indicate that patients with thyrotoxicosis treated with radioactive iodine in adult age have increased risk of developing parathyroid adenoma. PMID- 17118840 TI - Functioning thyroid carcinomas--some historical aspects. PMID- 17118841 TI - A prospective clinical trial to assess the efficacy of radioiodine ablation as an alternative to completion thyroidectomy in patients with differentiated thyroid cancer undergoing sub-total thyroidectomy. AB - We conducted a prospective clinical trial to evaluate whether radioiodine ablation can be an effective alternative to completion thyroidectomy in patients undergoing sub-total thyroidectomy and if yes, the optimum activity of 131I and frequency of ablation. A total of 85 patients (F--63; M--22) with mean age of 37.9+/-12.3 years were recruited in this study. The pre-ablation mean 24 hour radioiodine neck uptake, effective half-life, residual thyroid tissue weight and TSH values were 13.9+/-8.5%, 4.5+/-0.9 days, 9.6+/-3.6 g and 11.7+/-6.4 microIU/ml, respectively. Thyroid tissue was completely ablated in 50 patients (58.8%, 95% CI:50-68%) after mean 1st administered activity of 32.3+/-10.7 mCi of 131I and the cumulative ablation rate was 91.8% after two doses of 131I. During mean follow-up duration of 49 months no local/distant recurrence has been observed so far in this cohort. It appears that radioiodine ablation may be an attractive alternative to completion thyroidectomy and an activity as low as 35 mCi may achieve reasonable ablation. PMID- 17118842 TI - Glucocorticoid receptor subunit gene expression in thyroid gland and adenomas. AB - The present study was undertaken to investigate whether the glucocorticoid receptor -alpha (GR-alpha) and -beta (GR-beta) mRNA may be expressed in thyroid gland. Ten normal thyroid gland and 14 follicular adenomas were studied using a real-time fluorescent quantitative RT-PCR (FQ-RT-PCR) method. The results demonstrated that there was a lower expression of GR-alpha mRNA (x10(6) GR-alpha cDNA copies/microg total RNA) in thyroid adenoma (1.27+/-0.26) than that in normal thyroid gland (3.53+/-1.22) (p < 0.001). The expression of GR-beta mRNA was lower in all the thyroid tissues. Of note, there was a significant difference in GR-beta mRNA expression (x10(4) GR-beta cDNA copies/microg total RNA) between thyroid adenoma (80.8+/-13.9) and thyroid gland (1.78+/-0.59) (p < 0.001). The GR alpha/GR-beta ratios in thyroid adenoma and normal thyroid gland were 1.67+/-0.68 and 207.57+/-84.41 respectively (p < 0.001). These results revealed, for the first time, that both GR-alpha and GR-beta mRNA expression were detectable in both thyroid gland and adenomas tissues. We therefore conclude that down regulation of GR-alpha and up-regulation of GR-beta mRNA expression may play an important role in the thyroid adenomas. PMID- 17118843 TI - Short course continuous, hyperfractionated, accelerated radiation therapy (CHART) as preoperative treatment for rectal cancer. AB - Determine feasibility and toxicity of preoperative short course pelvic CHART (25 Gy in 15 fractions over 5 days) for treatment of clinically resectable primary rectal tumours. Between 1998 and 2004, 20 patients with clinically staged T3 resectable rectal carcinoma were treated in this prospective pilot study with preoperative short course CHART to their pelvis. The aim was for total mesorectal excision within 7 days. Radiation toxicity, surgical morbidity, locoregional control (LRC), overall (OS), cause specific (CSS) and disease free survival (DFS) outcomes were documented. Nineteen of the 20 patients completed planned radiotherapy. One discontinued radiotherapy due to toxicity. All patients underwent potentially curative radical surgery. One patient developed grade 3, and three patients grade 2 gastrointestinal toxicity. With a median follow-up of 31 months (range 0.9-88), there is no grade 3, 4 or 5 late toxicity. Two patients experienced grade 2, and three patients grade 1 late bowel toxicity. Two patients died from postoperative complications, and two developed grade 2 abdominal wound infections. At 3 years LRC is 95% (95% CI 83-100), OS 72% (95% CI 51-94), CSS 86% (95% CI 68-100) and DFS 80% (95% CI 60-100). Two patients died from metastatic disease, one patient from a second primary and one patient is alive after successful resection of hepatic metastases. This small study suggests preoperative short course CHART for clinically resectable rectal carcinoma is feasible with acceptable compliance and tolerable side effects. PMID- 17118844 TI - Effect of timing of surgery on survival after preoperative hyperfractionated accelerated radiotherapy (HART) for locally advanced rectal cancer (LARC): is it a matter of days? AB - We intend to analyse retrospectively whether the time interval ("gap duration" = GD) between preoperative radiotherapy and surgery in locally advanced rectal cancer (LARC) has an impact on overall survival (OS), cancer specific survival (CSS), disease free survival (DFS) and local control (LC). Two hundred seventy nine patients with LARC were entered in Trial 93-01 (hyperfractionated accelerated radiotherapy 41.6 Gy/26 Fx BID) shortly followed by surgery. From these 250 patients are fully assessable. The median GD of 5 days was used as a discriminator. The median follow-up for all patients was 39 months. GD > 5 days was a significant discriminator for actuarial 5-years OS (69% vs 47%, p = 0.002), CSS (82% vs 57%, p = 0.0007), DFS (62% vs 41%, p = 0.0003) but not for LC (93% vs 90%, p = non-significant). In multivariate analysis, the following factors independently predict outcome; for OS: age, GD, circumferential margin (CM) and nodal stage (ypN); for CSS: GD, ypN and vascular invasion (VI); for DFS: CEA, distance to anal verge, GD, ypN and VI; for LC: CM only. Gap duration predicts survival outcome but not local control. The patients submitted to surgery after a median delay of more than 5 days had a significantly better outcome. PMID- 17118845 TI - "Distributed proton radiation therapy"--a new concept for advanced competence support. AB - The increased interest in high precision radiation therapy is to a large extent driven by the potential of modern imaging technology. The aim of this project was to analyse how an expensive proton facility best could support a multi-centre health care system. We have developed a model for distributed expert collaboration where all clinical experts will work close to their patients in regional centres. Patients who are candidates for proton therapy will be examined and dose-planned at their regional clinic, discussed in a fully information supported video conference and digitally made available at the proton treatment facility. The proton facility itself will be placed near a communication centre easily reached by all patients where they will be treated under full responsibility of their own physician at the home clinic. This concept has been analysed in detail both with respect to the overall functionality and with respect to possible weaknesses. It was found that the concept of distributed radiation therapy, as proposed here, will offer a stable clinical solution for advanced radiation therapy. It will support the spread of knowledge, serve as a fully developed backup system and the concept will further serve as an efficient base for clinical research. PMID- 17118846 TI - Sequential evaluation of hepatic functional reserve by 99mTechnetium-galactosyl human serum albumin scintigraphy after proton beam therapy: a report of three cases and a review of the literatures. AB - The treatment strategy for malignant liver tumors should be appropriately determined because post-treatment quality of life greatly depends on the patients' residual hepatic function. In this report, we present three patients with malignant liver tumors treated by proton beam therapy in whom pre- and post therapeutic hepatic functional reserves were evaluated sequentially for more than a year by 99mTechnetium-galactosyl human serum albumin (99mTc-GSA) scintigraphy. All three patients exhibited the distinctive time course of 99mTc-GSA uptake efficiency, which suggested a transient decline in the ratio of liver activity to heart and liver activity at 15 minutes (LHL15) 3-6 months after proton beam therapy. This change was not in parallel with that expected from a functioning normal liver tissue volume. In a year after proton beam therapy, LHL15 recovered nearly to the pre-treatment level in all three patients. Our observations may be related to the up-regulation of receptor-mediated 99mTc-GSA uptake during hepatic regeneration after proton beam therapy. PMID- 17118847 TI - Clinical results of stereotactic body frame based fractionated radiation therapy for primary or metastatic thoracic tumors. AB - The aim of this study was to evaluate the treatment outcomes of stereotactic body radiation therapy for treating primary or metastatic thoracic tumors using a stereotactic body frame. Between January 1998 and February 2004, 101 lesions from 91 patients with thoracic tumors were prospectively reviewed. A dose of 10-12 Gy per fraction was given three to four times over consecutive days to a total dose of 30-48 Gy (median 40 Gy). The overall response rate was 82%, with 20 (22%) complete responses and 55 (60%) partial responses. The one- and two-year local progression free survival rates were 90% and 81%, respectively. The patients who received 48 Gy showed a better local tumor control than those who received less than 48 Gy (Fisher exact test; p = 0.004). No pulmonary complications greater than a RTOG toxicity criteria grade 2 were observed. The experience of stereotactic body frame based radiation therapy appears to be a safe and promising treatment modality for the local management of primary or metastatic lung tumors. The optimal total dose, fractionation schedule and treatment volume need to be determined after a further follow-up of these results. PMID- 17118848 TI - Correlation of COX-2 and K-ras expression to clinical outcome in gastric cancer. AB - Recent studies have suggested that K-ras play an important role in the induction of COX-2 expression in tumor cells. In the present study, tumor samples of 89 gastric cancer patients were prepared in tissue microarrays and they were stained by immunohistochemistry with antibodies against COX-2 and K-ras. We investigated the relationship between the protein expressions of COX-2 and K-ras in gastric cancer and their significance as prognostic markers in gastric cancer patients. The over expression rate of COX-2 and K-ras in gastric cancer was 61.8% and 61.8% (55/89) of all the patients, respectively. There was a significant positive correlation between COX-2 and K-ras expression in gastric cancer. COX-2 and K-ras positivity were correlated with depth of invasion and lymph node metastasis, respectively. K-ras positivity was correlated with growth pattern. Patients with COX-2 and K-ras positive tumors had a poorer prognosis than those with COX-2 and K-ras negative tumors. Over expression of COX-2 and K-ras were closely correlated to prognostic of patients with gastric cancer and they educed synergistic effect with carcinogenesis and development in gastric cancer. PMID- 17118849 TI - Treatment results of radiotherapy for carcinoma of the cervical esophagus. AB - The methods and results of treatment for cancer of the cervical esophagus differ from those for cancer of the thoracic esophagus. Our objective was to retrospectively review the outcome for cervical esophageal cancer patients treated with radiotherapy. Twenty-seven patients with carcinoma of the cervical esophagus treated with definitive radiotherapy from 1988 to 2002 were enrolled in the study. Clinical stage (UICC 1997) was stage I in five, II in six, III in 12 and IV in four. Concurrent head and neck malignancy was found in six patients (22%). The mean radiation dose was 66 Gy. Concurrent chemotherapy (cisplatin and 5-fluorouracil) was performed in 23 patients. The actuarial overall survival rates at 1, 3 and 5 years were 55.6%, 37.9% and 37.9%, respectively, with a median survival of 13.9 months. In the patients with stage I, the 3-year and 5 year survival rates were 75% and 75%, respectively. With univariate analysis, only two of the possible prognostic factors were found to actually influence survival: performance status (p < 0.01) and tumor length (p < 0.01). The survival of patients with cervical esophageal cancer remains poor. It is thought that organ preservation is possible by definitive chemoradiation for early cancer. PMID- 17118850 TI - Outcome and late complications of radiotherapy in patients with unicentric Castleman disease. AB - Castleman disease is a rare lymphoproliferative disorder. Surgery is considered standard therapy for the unicentric type. However, case reports have documented favorable responses to radiotherapy. The aim of this study was to analyse the clinical outcomes of five patients with unicentric Castleman disease treated with radiotherapy between 1991 and 2005. Mediastinal lymph nodes were the most common site of disease (four patients). Three patients were treated with radiotherapy alone, two patients with surgery and radiotherapy. Patients were treated with radiotherapy doses ranging from 40 Gy to 50 Gy. The median follow-up was 12 months (range, 3-175 months). During follow-up only one patient had progressive disease and died of Castleman disease. At the time of last follow-up two patients were in complete remission, one patient in partial remission, and one patient had stable disease. One patient showed serious acute and late toxicities. At the end of radiotherapy a paraneoplastic pemphigus vulgaris occurred, and eight to 11 months after radiotherapy a stenosis of the esophagus, of the left bronchus, and of the trachea due to scars. The study shows that unicentric Castleman disease is successfully treated with radiotherapy. However, for detection of possible complications as pemphigus vulgaris or stenosis of the esophagus or trachea an accurate follow-up is necessary. PMID- 17118851 TI - A patient surviving for eight years after proton and x-ray irradiation for advanced esophageal cancer. PMID- 17118852 TI - Behavioral disorders secondary to profound hypomagnesemia in a patient given cetuximab for metastatic colorectal cancer hypomagnesemia due to cetuximab treatment. PMID- 17118853 TI - Re-treatment with cetuximab in patients with severe hypersensitivity reactions to cetuximab. Two case reports. PMID- 17118856 TI - News from the Nordic colleges of general practitioners. The future is approaching. PMID- 17118857 TI - The Nordic congresses of general practice: a gateway to a global treasure? PMID- 17118858 TI - Prevention of disability by exercise among the elderly: a population-based, randomized, controlled trial. AB - OBJECTIVE: To assess the effectiveness of an intervention planned and implemented by regional geriatric care teams in order to prevent disability in an elderly population. DESIGN: Randomized, controlled trial with intention-to-treat and subgroup analyses. Subjects and setting. All home-dwelling persons aged 85 years or older in the City of Oulu (n = 555, 67%), including 486 persons with a history of recurrent falls or at least one risk factor for disability in the activities of daily living (ADL) or mobility. INTERVENTIONS: Programme consisting of home exercise, walking exercise, group exercise, and self-care exercise. MAIN OUTCOME MEASURES: Change in ADL and mobility during the intervention. RESULTS: Change in ADL performance did not differ (p = 0.462) but a positive change was found in mobility performance (p = 0.013) in the intervention subjects (n = 243) compared with the controls (n = 243). At the end of the intervention period, severe mobility restrictions existed similarly in intervention and control subjects: 48 (34%) and 46 (31%) (p = 0.650) respectively. Frequency of admissions into long term institutional care were similar in intervention and control groups: 15 (7%) and 13 (6%) (p = 0.669). Impaired balance existed in fewer intervention subjects (64, 45%) than controls (89, 59%) (p = 0.015). Frequency and times of home exercise activity reduced in control subjects (p < 0.001) but did not change in subjects assigned to home exercise intervention (p > 0.05). CONCLUSION: A pragmatic intervention slowed down the reduction of movement performance but positive effects could not be found in subjects with severe movement and any ADL dysfunction. Positive effects were also found in balance performance. PMID- 17118859 TI - Dietary and physical activity counselling on type 2 diabetes and impaired glucose tolerance by physicians and nurses in primary healthcare in Finland. AB - OBJECTIVE: To investigate the initiation of dietary and physical activity counselling and the arguments used when discussing physical activity and the type and consumption of dietary fats, during nurse-patient and physician-patient diabetic lifestyle counselling. DESIGN AND SUBJECTS: This study is a part of a larger follow-up research project focusing on diabetes counselling. The data include 129 videotaped counselling sessions between 17 patients and their physicians and nurses. Content analysis was carried out by identifying the verbal comments and reactions of participants concerning both physical activity and the type and consumption of dietary fats. RESULTS: The physicians and nurses spent little time on dietary and physical activity counselling. The counselling sessions consisted mainly of short questions with minimal feedback from either party. The beginning of the sessions concentrated on blood cholesterol levels and the extent of physical activity. The health professionals failed to emphasize the roles of reduced dietary fats and increased physical activity in substituting for or supplementing diabetes care. Although the patients communicated the factors that encouraged or discouraged physical activity the subject was generally not pursued by the health professionals. CONCLUSION: Within primary care there is a need for methods that would facilitate the discussion of health behaviour changes. In healthcare settings, education and training are required to develop the communication skills of health professionals in all aspects of preventive medicine. PMID- 17118860 TI - Abdominal obesity and insulin resistance in patients with type 2 diabetes in a Swedish community. Skaraborg hypertension and diabetes project. AB - OBJECTIVE: To explore the association between abdominal obesity and insulin resistance in patients with type 2 diabetes. DESIGN: A cross-sectional observational study. SETTING: Primary care in Skara, Sweden. SUBJECTS: A total of 198 men and 186 women with type 2 diabetes who consecutively completed an annual check-up in 1992-1993. MAIN OUTCOME MEASURES: Abdominal obesity was defined according to criteria for the metabolic syndrome using the waist circumference (WC): > 102 cm for men and > 88 cm for women. Insulin resistance was estimated using the Homeostasis Model Assessment (HOMA), and was dichotomized by the 75th percentile (IR). RESULTS: Abdominal obesity was found in 66 men (33%), and in 106 women (57%). Pearson's correlation coefficients between components of the metabolic syndrome and IR were statistically significant for WC, waist-hip ratio, serum triglycerides, and HDL cholesterol, and were higher for WC (0.40) than for waist-hip ratio (0.23) in both genders (p < 0.001). The association between WC and IR was challenged by successively entering other components of the metabolic syndrome into the model in a logistic regression. In the final model, adjusting for differences in age, systolic blood pressure, HbA1c, serum triglycerides, HDL cholesterol, and microalbuminuria, the association remained statistically significant both in men (OR 8.6, 95% CI 3.0-25.2, p < 0.001), and in women (OR 5.6, 95% CI 1.7-18.1, p = 0.004). CONCLUSIONS: WC provides a feasible measure for insulin resistance in the vast majority of subjects with type 2 diabetes. It is convenient and less expensive than direct means and could be used as a proxy for insulin resistance in population studies. PMID- 17118861 TI - Concepts underlying outcome measures in studies of consultations in general practice. AB - OBJECTIVE: To give an overview of the concepts used to describe and evaluate the outcome of general practice consultations. METHOD: A literature study was undertaken. Among 101 articles relevant to the subject 35 were chosen to illustrate the concepts found. RESULT: The following concepts were found: disease control, patient compliance, efficacy, symptom relief, enablement, general health, and patient satisfaction. The first three concepts encourage objective measurement while the following four concern the patient's subjective assessment. Methods naturally linked to the respective concepts are briefly described. DISCUSSION: The concepts reflect very diverse aspects of general practice. Some of the concepts come from other domains of research, while others have been developed within general practice. Disease is not exclusively in focus. The experience, competence, and life situation of the patient are as well dealt with as they should be in accordance with a patient-centred profession. However, the concepts spring from the researchers' ideas about what is important to patients. The patients' priorities were not sought. PMID- 17118862 TI - High blood pressure despite treatment: results from a cross-sectional primary healthcare-based study in southern Sweden. AB - OBJECTIVE: To study degree of blood pressure (BP) control in primary healthcare (PHC) treated hypertensive patients in relation to sex, age, drug treatment, and concomitant diseases. DESIGN: Random sample of patients with hypertension. SETTING: Ten PHC centres in the Region of Skane, Sweden. SUBJECTS: All the 30- to 95-year-old patients with hypertension who during the period 12 September to 24 September 2004 attended their PHC (146 men and 229 women). MAIN OUTCOME MEASURES: Achievement of BP control (< 140/90 mmHg) according to European guidelines. RESULTS: Some 90% had been treated > 12 months, 40% had mono-therapy, 15% > or = 3 drugs. Use of diuretics was more common in women while use of ACE inhibitors and calcium channel blockers was common in men. Inadequate BP control was related to age; only 22% had BP < 140/90 mmHg, 38% had a BP > or = 160/100 mmHg. BP decline was inversely related to BP measured 12 months or more prior to the present follow-up (r = - 0.64, p < 0.001, for systolic and r = - 0.67, p < 0.001, for diastolic BP). The systolic or diastolic BP had in every fifth patient during treatment increased by > or = 10 mmHg. No association was found between average BP decline and prescribed number of drugs. CONCLUSION: A minority of the patients had BP below the level (< 140/90 mmHg) recommended by European guidelines. This study illustrates the need for continued follow-up of defined groups of patients in order to improve quality of care. PMID- 17118863 TI - Primary care management of respiratory tract infections in Dutch preschool children. AB - OBJECTIVE: To determine age-specific antibiotic prescription and referral rates in preschool children diagnosed with acute respiratory tract infection (RTI) in primary care. DESIGN: Retrospective cohort study. SETTING: Research database of the Netherlands University Medical Center Utrecht Primary Care Network. SUBJECTS: All children aged 0-5 years within the database were included, covering 1998 to 2002 (18,801 child-years). MAIN OUTCOME MEASURES: Antibiotic prescription and referral rates were determined as percentage of children with at least one prescription or referral within a year, as total number of prescriptions per 1000 child-years, and as percentage of all RTI episodes. RESULTS: Antibiotics, mostly beta-lactam (80%) and macrolides (16%), were prescribed in 35% of RTI episodes. Annually 13% of the children received at least one antibiotic following an RTI. Antibiotics were prescribed in more than half of episodes of LRTI, sinusitis, AOM, and acute tonsillitis, and in 12-15% of episodes of asthma exacerbation, acute laryngitis, influenza acute, and acute upper respiratory infection (including common cold and pharyngitis). Almost 98% of RTIs were managed in primary care. On average 1% of the children were referred to a paediatrician or ENT specialist following RTI per year, especially after AOM (59% of referrals). Compared with older children, those under two years of age were more often treated with antibiotics (relative risk [RR] 1.4, 95% CI 1.3-1.6) and referred (RR 2.3; 95% CI 1.8-3.0). CONCLUSION: In the Netherlands most episodes of RTIs in preschool children were managed in primary care and this often involves prescription of antibiotics. Children younger than two years of age receive more often antibiotics for RTI and are also referred more, especially for AOM. PMID- 17118864 TI - Are patient characteristics helpful in recognizing mild COPD (GOLD I) in daily practice? AB - OBJECTIVE: To determine whether in a high-risk group of middle-aged male current smokers, patient characteristics are useful to recognize mild COPD (GOLD stage I). DESIGN: In a cross-sectional study spirometry was performed according to the American Thoracic Society criteria. COPD was defined according to the GOLD criteria for COPD. SETTING: Primary care. SUBJECTS: Male smokers, aged 40-65 years, without documented lung disease in the population at large. MAIN OUTCOME MEASURES: Medical records were scrutinized to collect patient characteristics. Multiple logistic regression analysis was used to identify independent determinants of mild COPD. RESULTS: A total of 567 subjects participated. COPD, defined by a FEV1/FVC ratio < 0.7, was detected in 170 subjects (30.0%, 95% CI 26.2-33.9%). In 149 subjects (26.3%; 22.7-30.1%) COPD was mild (GOLD stage I) and in 21 subjects (3.7%; 2.3-5.6%) moderate (GOLD stage II). Only age and cough were independently associated with the presence of mild COPD. The ability of these determinants to discriminate between subjects with or without mild COPD was relatively poor (ROC area 0.65). Therefore no prediction rule was developed. CONCLUSION: Our results indicate that patient characteristics are not helpful to recognize mild COPD (GOLD stage I) in middle-aged male smokers. PMID- 17118865 TI - Costs in primary care of investigating symptoms suspicious of cancer in a defined population. AB - OBJECTIVES: To calculate total cost of investigating symptoms related to one of the four most common cancers: prostate, breast, colorectal, and pulmonary. Special attention was given to two non-specific and common symptoms: vertigo/dizziness and tiredness/fatigue. DESIGN: Retrospective examination of patient records in a defined population. SETTING: Primary health care (PHC) and hospital care. MAIN OUTCOME MEASURES: Investigation costs of selected ICD codes. RESULTS: In total 6812 patients with 14,541 ICD codes were investigated to a total cost per registered ICD code, with one of the selected diagnostic measures, of SEK 800. Some 50% of the costs were due to X-ray examinations of which colonic X-ray was the most expensive single procedure. Symptoms related to colorectal cancer were the most expensive to investigate with vertigo/dizziness and tiredness/fatigue examined separately. Tiredness/fatigue symptoms cost SEK 395 and vertigo/dizziness SEK 197 per registered code to investigate and none of them yielded a detected malignancy. The incidence of cancer was 0.7%. The average cost of diagnosing one malignancy was SEK 236,700, if all diagnostic activities could be justified only for detection of malignancies. CONCLUSION: A high cost of diagnosing malignancy can be expected and justified, but many symptoms presented in PHC, such as tiredness/fatigue and vertigo/dizziness, have a relatively high investigation cost but detected no case of cancer in this study. PMID- 17118866 TI - Interviewing one's peers: methodological issues in a study of health professionals. AB - OBJECTIVE: Although health professionals are increasingly undertaking qualitative interviews with professional peers, there is little literature regarding the methodological implications of this process. The aim of the study was to elicit from informants their views on being interviewed by a fellow health professional. DESIGN: Semi-structured interviews with nine general practitioners (GPs), three rheumatologists, and three physical therapists, with a substantive focus on perceptions of osteoarthritis. The interviewer was a GP, and informants were asked for their reactions to being interviewed by a fellow professional. Data were analysed by hand, using a thematic approach. SETTING: Primary care clinics and practices in the UK. RESULTS: Although reassured to the contrary, many informants viewed the interview as a test of their professional knowledge. The interview was also seen by some GPs as serving an educational process, with the interviewer as an authoritative source of clinical information. There were some indications of professional vulnerability among informants in relation to possible scrutiny of their practice or knowledge, though none reported a negative experience of the interview. Notions of professional identity appeared central to many of the issues that emerged. CONCLUSION: The nature of the relationship in interviews involving professional peers creates specific methodological issues, which have important implications for qualitative research in primary healthcare. There are both advantages and disadvantages to interviewing professional peers, which should be considered in the light of the objectives of a particular study. PMID- 17118867 TI - Application of liquid chromatography-nuclear magnetic resonance spectroscopy for the identification of ethyldimethylpyrazine, a food flavouring agent. AB - The application of liquid chromatography-nuclear magnetic resonance spectroscopy (LC-NMR) for the direct identification of ethyldimethylpyrazine, a food flavouring agent, has been studied. The commercial product is a mixture of two regio-isomers, 2-ethyl-3,5-dimethylpyrazine (1) and 2-ethyl-3,6-dimethylpyrazine (2); however, the exact composition of the mixture is unknown. Structural characterization by LC-MS and GC-MS was not possible because both regio-isomers yield the same molecular related ion and ion fragmentation. To rapidly identify the two regio-isomers, the product was analyzed by LC-NMR with on-flow and fraction loop modes. From the results, the structure elucidations of the two regio-isomers could be carried out without the need to isolate the isomers by the usual procedures. PMID- 17118868 TI - Identification of hidden allergens: detection of pistachio traces in mortadella. AB - An analytical method based on the detection of specific DNA was developed and applied to mortadella samples with and without pistachio (Pistacia vera). The method is proposed for the detection of traces of pistachio deriving from previous processes or from accidental contamination, since in predisposed individuals pistachios can cause allergic reactions leading to anaphylactic shock. Three pairs of primers were identified and tested by polymerase chain reaction (PCR) on mortadella samples prepared with pistachio. Accidental contamination was also simulated. The optimized PCR was able to detect the presence of pistachio, even at low concentrations. The primers pair PSTC 1-2 is suggested for unambiguous identification of pistachio in mortadella. The limit of detection for this primers pair was 100 mg kg-1. No interference was observed from other spices or ingredients utilized in the formulation of the mortadella. The method enabled the identification of possible traces of pistachio remaining in the production plant after less than thorough washing. PMID- 17118869 TI - Usage of saccharin in food products and its intake by the population of Lucknow, India. AB - A survey of the usage patterns of the artificial sweetener, saccharin, in edible products and a study of its intake pattern in different population groups has been carried out. Of the different edible commodities, ice candy (87 samples) and crushed ice (14 samples), commonly consumed by children, and pan masala (16 samples) and pan flavourings (10 samples), consumed by the habitual population, were collected from different areas of Lucknow, India. Saccharin was extracted from the samples according to an AOAC method and analysed by HPLC. The consumption pattern of ice candy and crushed ice was determined for 6-20 year olds from a household dietary survey using the food frequency recall method (414 families having 1039 subjects). The consumption of pan masala and pan was assessed by a survey of habitual adult consumers comprising 782 and 1141 subjects, respectively. The average and maximum amounts of saccharin in pan masala samples were 12 750 and 24 300 mg kg-1, respectively, which are 1.6- and 3 fold higher than the maximum permitted levels allowed under Prevention of Food Adulteration (PFA) Act of India. In pan flavourings, the average and maximum amount of saccharin was 12.2 and 20.1%, i.e. 1.52- and 2.5-fold higher than the permissible limits of the PFA Act. The samples of ice candy and crushed ice showed average and maximum levels of 200 and 700 mg kg-1 and 280 and 460 mg kg-1, respectively. The average intake of saccharin through ice candy and crushed ice was less than 21% of the acceptable daily intake (ADI) (5 mg kg-1 body weight (bw) day-1). However, the maximum intake of saccharin, especially in the 6-10 year age group, contributed 57 and 68% of the ADI through ice candy and crushed ice, respectively. Maximum consumption of saccharin in all the age groups, if consuming both ice candy and crushed ice, results in exceeding the ADI by 54% for subjects in the 6-10-year age group. Hence, the 6-10-year age group population may be at risk of exceeding the ADI for saccharin. The average and maximum theoretical daily intake of saccharin through pan masala alone was 1.84 and 13.33 mg kg-1 bw day-1, contributing 37 and 267% of the ADI, whereas the estimated (maximum) daily intake was 810% of the ADI. The estimated maximum daily intake (EDI) of saccharin through pan was 6.87 mg kg-1 bw day-1, which was 137% of the ADI. Thus, individuals in the maximum consumption group for pan masala or pan may be susceptible to toxic effects of saccharin, including bladder distention, elevated urine osmolality and bladder cancer. PMID- 17118870 TI - Determination of acrylamide in coffee and coffee products by GC-MS using an improved SPE clean-up. AB - An improved gas chromatography-mass spectrometry (GC-MS) method to determine acrylamide (AA) in coffee and coffee products was developed. The method was based on two main purification steps: the first with ethanol and Carrez solutions in order to precipitate polysaccharides and proteins, respectively; and the second with a layered solid-phase extraction (SPE) column which proved to be efficient in the elimination of the main chromatographic interferences. The method is applicable to a wide range of coffee products. Twenty-six samples of different coffee products were analysed. The levels of AA were in the range 11.4-36.2 microg l-1 for 'espresso coffee' and 200.8-229.4 microg l-1 for coffee blends with cereals. The results indicate that the presence of cereals significantly increased the levels of AA. PMID- 17118871 TI - Levels of nitrate in Swedish lettuce and spinach over the past 10 years. AB - Monitoring of nitrate in Swedish-produced lettuce and spinach over the past 10 years (1996-2005) showed that more than 95% of the samples were below the maximum levels established by the European Commission in 1997. The good agricultural practices used by Swedish farmers may partly explain these results. Analytical results of organic farming production of lettuce from 2000 showed low nitrate levels compared with conventional production. The 10 years of Swedish experience has shown good compliance with the European Union maximum levels, but even lower nitrate levels may be achieved by organic farming methods, at least regarding fresh lettuce grown under cover. PMID- 17118872 TI - Fatty acid esters of 3-chloropropane-1,2-diol in edible oils. AB - A series of 25 virgin and refined edible oils, obtained from retailers, was analyzed for levels of free 3-chloropropane-1,2-diol (3-MCPD) and 3-MCPD released from esters with higher fatty acids (bound 3-MCPD). Oils containing free 3-MCPD ranging from <3 microg kg-1 (LOD) to 24 microg kg-1. Surprisingly, bound 3-MCPD levels were much higher and varied between <100 (LOD) and 2462 microg kg-1. On average, virgin oils had relatively low levels of bound 3-MCPD, ranging from <100 (LOD) to <300 microg kg-1 (LOQ). Higher levels of bound 3-MCPD were found in oils from roasted oilseeds (337 microg kg-1) and in the majority of refined oils (<300 2462 microg kg-1), including refined olive oils. In general, it appears that the formation of bound 3-MCPD in oils is linked to preliminary heat treatment of oilseeds and to the process of oil refining. Analysis of unrefined, de-gummed, bleached, and deodorized rapeseed oil showed that the level of bound MCPD decreased during the refining process. However, additional heating of seed oils for 30 min at temperatures ranging from 100 to 280 degrees C, and heating at 230 degrees C (260 degrees C) for up to 8 h, led to an increase in bound 3-MCPD levels. On the other hand, heating of olive oil resulted in a decrease in bound 3 MCPD levels. For comparison, fat isolated from salami was analyzed for intact fatty acid esters of 3-MCPD. This fat contained bound 3-MCPD at a level of 1670 microg kg-1 and the fatty acid esters of 3-MCPD mainly consisted of 3-MCPD diesters; monoesters of 3-MCPD were present in smaller amounts. The major types of 3-MCPD diesters (about 85%) were mixed diesters of palmitic acid with C18 fatty acids (stearic, oleic, linoleic acids). These diesters were followed by 3 MCPD distearate (11%) and 3-MCPD dipalmitate (4%). Generally, very little 3-MCPD existed as the free compound (31 microg kg-1). PMID- 17118873 TI - Quantification of seven arsenic compounds in seafood products by liquid chromatography/electrospray ionization-single quadrupole mass spectrometry (LC/ESI-MS). AB - A liquid chromatography/electrospray ionization-single quadrupole mass spectrometry (LC/ESI-MS) method was developed to quantify seven arsenic compounds: arsenate (As(V)), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), arsenobetaine (AB), trimethylarsine oxide (TMAO), arsenocholine (AC) and tetramethylarsonium ion (TEMA), widely found in seafood. The arsenicals separated by anion- or cation-exchange LC were all readily identified under the optimized ESI-MS conditions. Linear calibration curves constructed by plotting the peak area counts of molecular ions against the arsenic concentrations were obtained for all seven arsenic compounds. The limits of quantification (S/N = 10) were 800, 600, 50, 10, 5, 5 and 5 ng ml-1 for As(V), MMA, DMA, AB, TMAO, AC and TEMA, respectively. The LC/ESI-MS method was found to be useful to quantify arsenic compounds in seafood by model experiments using the mid-gut gland and muscle of a shellfish (Buccinid whelks). Spiking experiments verified the accuracy of the method. PMID- 17118874 TI - Quantitative analysis of ochratoxin A in wine and beer using solid phase extraction and high performance liquid chromatography-fluorescence detection. AB - A reliable and accurate method is described for the quantitative analysis of ochratoxin A (OTA) in wine and beer. The method involves the use of disposable non-polar polymeric and aminopropyl solid-phase extraction cartridges to isolate the mycotoxin from alcoholic beverages. Extracts were subsequently analysed using reverse-phase high-performance liquid chromatography-fluorescence detection with post column ammoniation to improve the limit of detection. The precision of the method determined at three levels in both wine and beer was less than 5% (RSD). Standard addition studies in both wine and beer showed that the recovery of OTA varied between 90 and 106% over a concentration range of 0.016-1.284 microg l-1. The detection and quantification limits were shown to be better than 0.004 (S/N = 3) and 0.016 microg l-1 (S/N = 10) for both beer and wine. PMID- 17118875 TI - Patulin distribution in Fuji and Golden apples contaminated with Penicillium expansum. AB - This work assesses the extent of patulin contamination in Penicillium expansum infected apples stored at room temperature for short periods of time and its relationship with apple variety (Golden or Fuji), degree of ripeness and size of lesions. Inoculated apples were incubated at 20 degrees C. Patulin was determined in both sound and decayed tissue from cylindrical samples taken around the lesions and cut into 0.5-cm thick sections. Higher accumulation of patulin occurred in Golden apples, with less ripened apples showing higher concentrations. Total accumulated patulin was similar or higher in 4-cm compared to 2-cm lesioned apples, although a decrease in patulin concentration was observed in older lesion sections. Patulin accumulation occurred over a short period of time at room temperature, thus the stand-by period before processing should be minimised. Of total patulin, 2-6% migrated to the surrounding sound tissue, thus trimming tissue around the rotten part may be a good preventive practice for apple derivative production. PMID- 17118876 TI - Occurrence of polybrominated diphenyl ethers and polychlorinated biphenyls in shark liver oil supplements. AB - Results are reported of a pilot survey of concentrations of polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) in shark liver oil supplements. Eleven brands of dietary supplements were analysed using an isotope dilution GC/MS method. Total concentrations of 10 PBDE congeners (BDE-28, -47, 49, -66, -99, -100, -153, -154, -155 and -183) ranged from 0.1 to 53 ng g-1 oil weight and total concentrations of six PCB congeners (CB-28/31, -52, -118, -153 and -180) in the samples ranged from 16 to 340 ng g-1 oil weight (undetected values are not included). Two brands of Japanese deep-sea shark liver oil contained the highest levels of PBDEs (49-53 ng g-1 oil weight) and PCBs (290-340 ng g-1 oil weight). These results indicate that PBDEs may have entered Japanese deep-sea waters. PMID- 17118877 TI - Effect of alginate composition and gelling cation on microbead swelling. AB - PURPOSE: The purpose of this study was to determine the roles of alginate composition and gelling cations on bead swelling, which affects its durability. METHOD: Using a 2-channel droplet generator, microspheres were generated with 1.5% solutions of low viscosity high-mannuronic acid (LVM), medium viscosity high mannuronic acid (MVM), low viscosity high-guluronic acid (LVG) and medium viscosity high-guluronic acid (MVG) alginate. They were gelled by cross-linking with 1.1% solution of either BaCl2 or CaCl2. The diameters of the microbeads were measured and recorded on day 0. The microbeads were subsequently washed and incubated in saline at 37 degrees C for 2 weeks with size assessment every 2 days. The data were normalized by calculation of the percentage change from control (day 0) for all groups of microbeads. RESULTS: Diameters of all beads were between 550-700 microns on day 0. Viscosity had no effect on swelling of Ba++- and Ca++-alginate microbeads. Ca++-alginate microbeads were more prone to swelling than the corresponding Ba++-alginate beads. High G-Ba++ beads had only a modest increase in size over time, in contrast to the high M-Ba++. CONCLUSION: Alginate composition and the gelling cation have significant effects on bead swelling. PMID- 17118878 TI - Preparation and control-release kinetics of isosorbide dinitrate microspheres. AB - Microcapsules for sustained release of poorly soluble isosorbide dinitrate (ISDN) were prepared based on ethylcellulose (EC) and/or blended with appropriate amounts of relatively hydrophilic hydroxypropyl cellulose (HPC) as matrix materials using the oil-in-oil emulsion evaporation method. The microspheres studied had three-mode sizes (100-150, 250-300 and 400-450 microm) and four polymer compositions (1, 0.833, 0.67 and 0.5 weight fraction EC). The microspheres were observed to contain essentially no drug crystalline domain and were of a porous morphology. The cumulative amounts of ISDN releasing from the microspheres as functions of mode fractions size and polymer compositions were measured in vitro. It was observed that the microspheres' size influenced the release behaviour of drug more obviously than the polymer composition. The smaller size and the higher hydrophilic HPC content show the faster release rate of drug and the smaller amount of drug residue. The kinetics of drug release depends on the size and polymer composition. The microspheres with 100-150 microm, of all polymer compositions, present one-stage diffusion kinetic with a lag period for drug release. On the other hand, the microspheres with the other two sizes exhibit two-stage diffusion kinetic with a lag period. According to the kinetic model, the microspheres obtained are surmised to have a core-shell like drug concentration distribution and/or a core-shell morphology. PMID- 17118879 TI - Formulation and testing of vancomycin loaded albumin microspheres prepared by spray-drying. AB - Microparticles are widely employed as carriers of biologically active compounds with many possible applications. For targeted drug delivery and sustained release purposes, biopolymers (i.e. polysaccharides and proteins) have been proposed. In this study, microsphere formulations of vancomycin were prepared by the spray drying method. Bovine serum albumin (BSA) was used as a polymer matrix and was cross-linked with glutaraldehyde after microsphere preparation. The product yield obtained from the spray-drying method was approximately 75%. The mean particle size was 5 +/- 1.6 microm, with the majority of particles between 4 and 8 microm. The extent of cross-linking affected the release of vancomycin from microspheres. Moreover, both rate and extent of vancomycin release from microspheres decreased with increasing glutaraldehyde concentration. Encapsulation of vancomycin did not alter the bioactivity of the drug and it was more effective in killing Staphylococcus aureus than the solution form. PMID- 17118880 TI - Characterization and in vitro degradation of salicylate-derived poly(anhydride ester microspheres). AB - The aim of this study was to investigate how glass transition temperature (Tg) influenced polymer microsphere formation and degradation of three chemically, similar novel salicylatebased poly(anhydride-esters): poly[1,6-bis(o carboxyphenoxy)hexanoate] (CPH), Tg = 59 degrees C; poly[1,8-bis(o carboxyphenoxy)octanoate] (CPO), Tg = 30 degrees C; and poly[1,10 bis(ocarboxyphenoxy) decanoate] (CPD), Tg = 27 degrees C. Microspheres of these polymers were prepared using a modified oil-in-water solvent evaporation method and processed by either resuspension or washed by centrifugation. The morphology of the microspheres determined by scanning electron microscopy (SEM) revealed that an extra washing step appears to increase aggregation as the Tg decreases; whereas only limited aggregation occurred in the polymer with the lowest Tg, CPD, in those not washed by centrifugation. Residual polyvinyl alcohol apparently affected the drug release rates from the microspheres by a stabilization process that produced an 8 h lag time and a 5% decrease in the amount of drug released over a 7 day period compared to microspheres washed free of PVA. These results demonstrate that salicylate-based poly(anhydride-esters) with sufficiently high Tgs, can be processed into microspheres that release salicylate over a time period amenable for drug delivery applications. PMID- 17118881 TI - New microencapsulation system for ascorbic acid using pea protein concentrate as coat protector. AB - Microencapsulation is essential to preserve biological activity of ascorbic acid (AA) and pea protein has not been used as a carrier in such processes. This work aimed to produce microparticles by a spray-drying process using pea protein (PPC) as wall material of AA and evaluate the retention of the core by HPLC, overall morphology SEM, size distribution by light scattering and release kinetics. Carboxymethylcellulose (CMC) and blends with maltodextrin (M) were produced for comparative analyses. The yields were compatible with the applied technology and the retention was above 84% for all materials. The PPC microparticles presented irregular and rough surfaces, CMC produced a regular and smooth surface and agglomeration was more intense in microparticles with M. Mean particle diameters were all below 8 microm. The microparticle release rates were lower than those with free AA, being best correlated to the Higuchi kinetic model. These results support the utilization of PPC for microencapsulation of AA. PMID- 17118882 TI - Encapsulation of RIN-m5F cells within Ba2+ cross-linked alginate beads affects proliferation and insulin secretion. AB - The viability, proliferation and insulin production of RIN-m5F cells when loaded into alginate beads to form a 3D culture system has been investigated. The mechanism of alginate cross-linking (calcium ions vs barium ions), the addition of poly(L-lysine) (PLL) and poly(L-ornithine (PLO) and presence of different extra-cellular matrix proteins (ECM) influence the RIN-m5F cell behaviour. Cells in calcium alginate beads (CAB) proliferated and produced more insulin per cell than monolayer culture, but the physical properties of the beads were poor and they ruptured within a few days of culture. Barium alginate beads (BABs) provided a stable encapsulation method. Addition of PLL and PLO at concentrations above 0.1% w/v with the culture medium increased cell proliferation. With the addition of ECMs after bead formation there was a further increase in cell proliferation for certain combinations of ECM and PLO. It was concluded that RIN-m5F-loaded Ba alginate beads (BABs), when incorporated with varying concentrations of poly (L) lysine (PLL), poly (L) ornithine (PLO) in the presence of extra-cellular matrix proteins (ECMs) were superior to both tissue culture and RIN-m5F-loaded Ca alginate beads (CABs) in terms of physical stability, cell proliferation and insulin production. PMID- 17118883 TI - Preparation and release of model drugs from thermally sensitive poly(N isopropylacrylamide) based macrospheres. AB - Emulsion polymerization was employed to prepare poly(N-isopropylacrylamide) hydrogel spheres, which exhibited an LCST of 32 degrees C. The hydrogels were loaded with model drugs (benzoic acid (BA), sodium benzoate and diltiazem HCl (DHCl)) and release investigated at 25 degrees C and 37 degrees C. The temperature at which gel formation occurred was vital for successful hydrogel preparation, macrosphere formation not occurring when the temperature was close to the LCST. Sphere size increased on decreasing the stirring rate and on slowing the rate of addition of the aqueous phase. Pulsatile delivery was investigated using BA and DHCl. For both compounds a pulse was observed with a change in temperature. Pulsed release of the smaller model drug of lowest solubility, BA, was more successful. Drug release from hydrogel spheres was, therefore, found to be dependent on the physicochemical properties of the drugs, with pulsatile release of low molecular weight compounds, by temperature cycling, difficult to control. PMID- 17118884 TI - Microspheres from new biodegradable poly(ester amide)s with different ratios of L and D-alanine for controlled drug delivery. AB - A series of biodegradable poly (ester amide)s composed of sebacic acid, dodecanediol and different ratios of the stereoisomers of L- and D-alanine were synthesized for applications in drug delivery systems. Microspheres loaded with diclofenac sodium salt, triclosan and clofazimine were prepared with the solvent evaporation technique. No influence of polymer constitution in the drug release rate was found in vitro and no degradation occurred during the period of drug release. It was shown that a sustained delivery of the hydrophilic diclofenac sodium salt in Sorensen media occurred and it was controlled by diffusion. However, exhaustion of microspheres was feasible only from the most porous matrices where channelling had an important contribution. PMID- 17118885 TI - Preparation and characterization of biodegradable urea-loaded microparticles as an approach for transdermal delivery. AB - This work describes the formulation and characterization of urea-loaded microspheres prepared using various polymers such as ethyl cellulose (EC), cellulose acetate phthalate (CAP) and poly (D,L-lactic-co-glycolic acid) (PLGA), along with the utilization of a solvent evaporation technique. The effect of various formulation parameters (i.e. polymer type and concentration, vehicle type, polymer solution/vehicle volume ratio, drug/polymer ratio, homogenizer and stirrer speed, sonication time and speed, type of washing solution, drying and separation method) on the characteristics of microspheres was also evaluated. Results obtained indicated that, in the presence of urea, highest rate of EC microsphere production could be obtained at a drug/polymer ratio of 1:2 and a polymer solution/vehicle volume ratio of 1:50. In some cases, crystallization of urea was observed during the encapsulation process using cellulose derivative polymers. CAP microparticles showed a rough and tortuous surface while EC microparticles had a wider range of particle size. However, with the PLGA polymer, much better desired microparticles with a smaller size range of 1-3 microm were obtained. In general, PLGA microspheres were spherical in shape and possessed smooth surfaces with less pores in comparison with those obtained by the other polymers. The yield of particle production and the extent of urea encapsulation in PLGA particles were measured to be 68.87% +/- 5.3 and 40.5% +/- 3.4, respectively. The release study from PLGA microspheres revealed that up to 70% of the drug was released within a few days, through a four-stage release pattern. PMID- 17118887 TI - Literature alerts. PMID- 17118889 TI - Hypoxia: importance in tumor biology, noninvasive measurement by imaging, and value of its measurement in the management of cancer therapy. AB - PURPOSE: The Cancer Imaging Program of the National Cancer Institute convened a workshop to assess the current status of hypoxia imaging, to assess what is known about the biology of hypoxia as it relates to cancer and cancer therapy, and to define clinical scenarios in which in vivo hypoxia imaging could prove valuable. RESULTS: Hypoxia, or low oxygenation, has emerged as an important factor in tumor biology and response to cancer treatment. It has been correlated with angiogenesis, tumor aggressiveness, local recurrence, and metastasis, and it appears to be a prognostic factor for several cancers, including those of the cervix, head and neck, prostate, pancreas, and brain. The relationship between tumor oxygenation and response to radiation therapy has been well established, but hypoxia also affects and is affected by some chemotherapeutic agents. Although hypoxia is an important aspect of tumor physiology and response to treatment, the lack of simple and efficient methods to measure and image oxygenation hampers further understanding and limits their prognostic usefulness. There is no gold standard for measuring hypoxia; Eppendorf measurement of pO(2) has been used, but this method is invasive. Recent studies have focused on molecular markers of hypoxia, such as hypoxia inducible factor 1 (HIF-1) and carbonic anhydrase isozyme IX (CA-IX), and on developing noninvasive imaging techniques. CONCLUSIONS: This workshop yielded recommendations on using hypoxia measurement to identify patients who would respond best to radiation therapy, which would improve treatment planning. This represents a narrow focus, as hypoxia measurement might also prove useful in drug development and in increasing our understanding of tumor biology. PMID- 17118890 TI - 'Being powerful beyond measure': lessons from Africa about the global practice of physical therapy in the 21st century. AB - In part, this is a synopsis of a talk Professor Dean was invited to share with the physical therapy students at the University of Nigeria at Enugu in July 2005. At that time, the students honored her with an Igbo name, 'Chinere' - a true highlight of her career. PMID- 17118891 TI - The effects of gender, age, and body mass index on standing lumbar curvature in persons without current low back pain. AB - Reference values for standing lumbar curvature (SLC) obtained via noninvasive methods are not well established in persons without current low back pain. The effect of gender is considered to have a significant effect on SLC with women having more lumbar lordosis than men. The effect of age and degree of obesity are not considered to have a statistically significant effect on SLC. The purpose of this study was to test the assumption that measurements of SLC in healthy adults obtained by a flexible curve will differ between genders, whereas the SLC will not differ across categories of age and body mass index (BMI). Two hundred thirty five volunteers (119 men and 116 women) whose ages ranged between 20 and 79 years participated in the study. Subjects were almost exclusively White and from the Midwest. Measurements of the SLC were obtained by a flexible curve. The curve's shape was transferred to poster board, and the value of SLC was quantified by a previously described technique. A three-way analysis of variance (alpha = 0.05) was used to examine the main effects of gender, age, and BMI on SLC. The effect of gender (F1,199 = 21.4, p < 0.0001) and the effect of age (F5,199 = 2.8, p < 0.017) were statistically significant. The effect of BMI (F2,199 = 1.8, p = 0.176) was not significant. Women (mean, 49.5 degrees +/-10.7 degrees ) demonstrated about 6.5 degrees more SLC than their male (mean, 43.0 degrees +/ 10.7 degrees ) counterparts. For age, the only significant difference was between the 20 to 29- and 50 to 59-year-old age categories. This study provides physical therapists with typical values of SLC in men and women without current low back pain. PMID- 17118892 TI - Role of conceptual models in a physical therapy curriculum: application of an integrated model of theory, research, and clinical practice. AB - The Department of Physical Therapy, University of Alberta, Edmonton, Alberta, Canada, recently implemented a Master of Physical Therapy (MPT) entry-level degree program. As part of the curriculum design, two models were developed, a Model of Best Practice and the Clinical Decision-Making Model. Both models incorporate four key concepts of the new curriculum: 1) the concept that theory, research, and clinical practice are interdependent and inform each other; 2) the importance of client-centered practice; 3) the terminology and philosophical framework of the World Health Organization's International Classification of Functioning, Disability, and Health; and 4) the importance of evidence-based practice. In this article the general purposes of models for learning are described; the two models developed for the MPT program are described; and examples of their use with curriculum design and teaching are provided. Our experiences with both the development and use of models of practice have been positive. The models have provided both faculty and students with a simple, systematic structured framework to organize teaching and learning in the MPT program. PMID- 17118893 TI - A functional model to describe the action of the adductor muscles at the hip in the transverse plane. AB - Anatomical texts agree on most muscle actions, with a notable exception being the action of the adductors of the hip in the transverse plane. Some texts list an action of the adductor brevis (AB), adductor longus (AL), and/or adductor magnus (AM) as internal rotation, whereas others list an action of external rotation. The purpose of this article is to present a functional model in support of the action of external rotation. Transverse plane motion of the femur at the hip during normal gait is driven by subtalar joint motion during the loading response, terminal stance, and preswing phases. During the loading response, the subtalar joint pronates, and the talus adducts. This talar adduction results in the lower leg, and subsequently the femur, internally rotating. During terminal stance and preswing, the opposite occurs; the subtalar joint supinates as the talus abducts in response to forces generated from the lower extremity and in the forefoot. Electromyographic (EMG) studies indicate varied activity in the AB, AL, and AM during the loading response, terminal stance, and preswing phases of the gait cycle. A careful analysis of EMG activity and kinematics during gait suggests that, in the transverse plane, the adductors may be eccentrically controlling internal rotation of the femur at the hip during the loading response, rather than the previously reported role as concentric internal rotators. In addition, these muscles may also concentrically produce external rotation of the femur at the hip during terminal stance and preswing. Physical therapists should consider this important function of the hip adductors during gait when evaluating a patient and designing an intervention program. Anatomical texts should consider listing the concentric action of external rotation of the femur at the hip as one action of the AB, AL, and AM, particularly when starting from the anatomic position. PMID- 17118894 TI - Descriptors of American Physical Therapy Association physical therapist members' reading of professional publications. AB - One of the components of evidence-based practice (EBP) is reading the literature. The purpose of this investigation was 1) to determine which publications are read most frequently by physical therapists (PTs), 2) to identify employment and education characteristics related to reading patterns, and 3) to determine how PTs use information gained from reading professional publications. A survey was constructed, pretested, revised, and then mailed to a random sample of 1,000 physical therapists, 500 from the general American Physical Therapy Association (APTA) membership and 500 from the combined Geriatric and Orthopedic section memberships. The data were analyzed with descriptive statistics. After all mailings, 43.3% of the sample responded. Of those responding, 66.3% were employed as clinicians, 14.7% as clinician-administrators, 6% as educators, and 4.9% as administrators. Overall, and for clinicians, the two most frequently read professional publications were non peer-reviewed. For clinician groups "patient management" was among the top two most selected uses of information from reading professional publications. Educators and those with an advanced doctorate indicated "class lectures" as either their second most frequently selected use or tied for first with "keeping current." Only educators and those with or pursuing an advanced doctorate reported using information from reading professional publications for "research ideas" or "research methods" among their top five uses. Slightly more than 10% of the respondents cited a peer-reviewed published article as having been most influential on their practice. Non peer-reviewed professional publications appear to serve as a more frequent source of information for "patient management" than do peer-reviewed publications. Efforts to increase the use of EBP need to be explored and evaluated for impact on physical therapists' practice. PMID- 17118895 TI - Effective intervention strategies for management of impaired posture and fatigue with post-polio syndrome: a case report. AB - This case report describes effective intervention strategies that included gait training with the use of an acceptable, assistive device for the management of impaired posture and fatigue associated with post-polio syndrome. Review of videotaped gait-training sessions enhanced patient compliance with the assistive device, a single forearm crutch. The 49-year-old female acquired childhood polio with resulting leg length discrepancy, lower extremity weakness, and complaints of fatigue. She walked with a right ankle foot orthosis and increased right lateral trunk flexion during right stance. Interventions consisted of patient education regarding the diagnosis and management of post-polio syndrome, health promotion, and energy conservation strategies that included gait training with a single forearm crutch. An outcome measure not previously documented in the post polio literature was used to measure the patient's perceived fatigue. Six-month and 18-month reexaminations found that the patient was compliant with the assistive device, reported a 30% reduction in fatigue, and walked with a more upright posture for longer distances in a shorter period of time. Patient education, health promotion, and energy conservation strategies that included walking with a properly fitting assistive device reduced perceived fatigue and improved posture and function in an individual with post-polio syndrome. PMID- 17118896 TI - End-to-side nerve repair induces nuclear translocation of activating transcription factor 3. AB - We wanted to find out if any of three different types of manipulations: a piece of muscle or nerve put parallel to a nerve; an epineurial window made, or sutures inserted into a nerve, or both; or pieces of nerve sutured to an epineurial window end-to-side to the musculocutaneous or sciatic nerve, resulted in activation of activating transcription factor 3 (ATF3) in neurons and in non neuronal cells. ATF3, a marker of cell activation, was investigated by immunocytochemistry one week after manipulation. A piece of nerve or muscle parallel to a nerve did not induce ATF3 locally in the nerve and induced ATF3 only rarely in neurons. In contrast, an epineurial window or insertion of sutures, or both, with or without attachment of a piece of nerve placed end-to side, induced robust ATF3 expression. We conclude that an injury to a peripheral nerve trunk associated with end-to-side nerve repair, activates neurons and non neuronal cells and may contribute to sprouting of axons into the nerve attached end-to-side. PMID- 17118897 TI - Effectiveness of short-contact topical tretinoin in promoting wound healing in db/db mice. AB - We have investigated the effects of short-contact topical application of tretinoin using 16 male db/db mice by creating two full-thickness wounds on the skin of the back. A 0.1% tretinoin aqueous gel was applied to one wound for five minutes daily for five successive days while only aqueous gel was applied to the other (control). The mean (SD) percentage surface area unhealed before and after treatment were 0.88 (0.3) and 0.64 (0.15). In tretinoin-treated mice and non treated mice (controls), the mean (SD) thicknesses of granulation were 1.383 (697) micro m and 683 (413) micro m, the density of capillary vessels in granulation was 12.2 (5.5)% and 5.7 (3.9)%, respectively. Differences between the two groups were significant for each variable. Wound healing was accelerated with short-contact topical application of tretinoin in db/db mice. PMID- 17118898 TI - New digital method for quantitative assessment of nasal morphology. AB - Our aim was to develop and validate a new method to assess objectively and quantitatively the morphology of the nostrils after nasal or nasolabial surgery. We used digital analysis using specific mathematical algorithms to assess several geometric measurements, particularly of facial asymmetry, expressed in adimensional units. Forty-five patients with no facial anomalies (control group) were used initially to evaluate the method and to obtain variables for statistical reference. Thirty-five patients operated on for unilateral cleft lip and palate (cleft group) were then analysed and compared with the control group. Individual scores were obtained for each patient, computed, and correlated with those established by a lay panel. Statistical analysis showed good sensitivity and reliability (R>0.8). PMID- 17118899 TI - Study of changes in nasal morphology in children with complete unilateral cleft lip and palate after secondary bone grafting measured with image analysis. AB - We aimed to evaluate retrospectively changes in the nasal appearance after secondary bone grafting in children with complete unilateral cleft lip and palate in 41 consecutive patients. Basal view photographs were analysed by a software program (ASYMNOS, for asymmetry of nostrils, available on simple request) that computes comparative functions between both nostrils and gives an objective score. These data were compared with those of a control group. The measurements and scores were calculated for each group, and then the global objective scores were correlated with the subjective scores that resulted from an independent external panel that ranked the aesthetic aspects of the nostrils. There was a significant improvement in the nasal symmetry postoperatively that resulted from a decrease in height between the nostrils, and a positive correlation between the objective and subjective scores. PMID- 17118900 TI - Waste of skin in elliptical excision biopsy of non-melanomatous skin cancer. AB - Our objective was to quantify the skin wasted during elliptical excision biopsy. This is defined as the difference between the excised elliptical area and the area of the original lesion. We used geometrical calculations of 26 excisional biopsy specimens from patients with non-melanomatous malignant tumours. Relative to the lesion area, the largest waste of skin measured was 230% and the smallest was 34% with a mean of 130%. We conclude that for better preservation of tissue when closing circular defects, patterns other than an ellipse should be adopted. PMID- 17118901 TI - Fibroma of the tendon sheath that expanded into the radiocarpal joint with bony involvement. AB - We present a 46-year-old man with a fibroma of the tendon sheath of the left radiocarpal joint and bony involvement of the scaphoid and the radial styloid. Bony involvement of a fibroma of the tendon sheath is rare and has been reported only four times to our knowledge. PMID- 17118902 TI - Middle phalangectomy: a functional and aesthetic cure for macrodactyly. AB - Macrodactyly is a rare congenital anomaly of the extremities. Some treatments such as debulking, epiphysiodesis, stripping or resection of the digital nerve, shortening of the fingers, amputation, and angulation osteotomy have been tried. We describe a patient with a giant digit in the hand, and present middle phalangectomy as a suitable treatment. PMID- 17118905 TI - Frequency-compression outcomes in listeners with steeply sloping audiograms. AB - Previous investigation of an experimental, wearable frequency-compression hearing aid revealed improvements in speech perception for a group of listeners with moderately sloping audiograms (Simpson et al, 2005). In the frequency-compression hearing aid, high frequencies (above 1600 Hz) were amplified in addition to being lowered in frequency. Lower frequencies were amplified without frequency shifting. In the present study, an identical frequency-compression scheme was evaluated in a group of seven subjects, all of whom had steeply sloping hearing losses. No significant differences in group mean scores were found between the frequency-compression device and a conventional hearing instrument for understanding speech in quiet. Testing in noise showed improvements for the frequency-compression scheme for only one of the five subjects tested. Subjectively, all but one of the subjects preferred the sound quality of the conventional hearing instruments. In conclusion, the experimental frequency compression scheme provided only limited benefit to these listeners with steeply sloping hearing losses. PMID- 17118906 TI - SII and fit-to-target analysis of compression system performance as a function of number of compression channels. AB - This work was undertaken to answer the question, 'How does the speech audibility/fit-to-gain-target provided by compression change with number of channels?' For each of 957 audiograms and a given number of compression channels, the channel crossover frequencies were set either to maximize the SII (speech intelligibility index) for low- and high-level speech spectra, or to optimize the fit-to-gain targets from the Cambridge method for loudness equalization (CAMEQ). The audiograms comprised all common configurations, and losses ranged from mild to severe. Use of these computational procedures allowed the predicted, channel number-based performance to be determined separately from the effects of other compression parameters. From one to five channels were sufficient to yield predicted speech recognition performance within 5% of maximum for 90% of the 'mild' and 'moderate' audiograms. Three to nine channels were necessary for the same level of predicted performance for 90% of the 'severe' audiograms. Four channels or fewer were sufficient to produce less than 5 dB rms error in fit to CAMEQ targets for 90% of all audiograms. PMID- 17118907 TI - Auditory processing tests for Spanish-speaking adults: an initial study. AB - There is no standardized material to assess auditory processing (AP) in Spanish speaking adults. The purpose of this research was to develop and obtain initial normative data for AP tests for Spanish-speaking adults. The AP tests comprised the Hong Kong CAPA (Hong Kong central auditory processing assessment) including speech-in-noise, binaural fusion, filtered speech, and dichotic digit tests. In addition five tests already developed by other authors, but with no normative data reported for the target population, were included in the study. These tests were staggered spondaic words, random gap detection, pitch pattern sequence, duration pattern sequence, and masking level difference. Forty normal hearing Chilean adults were assessed to obtain the initial normative data. Several correlations were found between tests and between subtests within the same test category. 90th percentiles are suggested as initial normative data. PMID- 17118908 TI - Prevalence of hearing loss in 1598 adults with an intellectual disability: cross sectional population based study. AB - A cross-sectional epidemiological study on prevalence of hearing loss was carried out in an age- and Down's syndrome- stratified random sample of 1598 persons drawn from a base population of 9012 persons, representative of the Dutch adult population of intellectual disability (ID) service users. The re-weighted population prevalence is 30.3% (95% confidence interval [CI]: 27.7-33.0%). Subgroup prevalences range from 7.5% (95% confidence interval [CI]: 3.6-13.3) in the subgroup aged 18-30 years with ID by other causes than Down's syndrome, up to 100% (95% CI: 79.4-100%) in adults over 60 years of age with Down's syndrome. Down's syndrome (OR 5.18, 95% CI 3.80-7.07) and age were confirmed to be risk factors. Age-related increase in prevalence in persons with Down's syndrome appears to occur approximately three decades earlier, and in persons with ID by other causes approximately one decade earlier than in the general population. PMID- 17118909 TI - Prevalence of cochlear implants in postlingually deafened adults in a Swedish region. AB - Cochlear implantation (CI) rates vary between countries, depending on identification routines and economic restrictions. The present study aimed at determining the prevalence of CIs in postlingually deafened, aged 20-69 years old, in Goteborg, Sweden. Three patient databases with information on PTA, a questionnaire, medical records and consultations identified 88 subjects with sensorineural hearing loss >/=80 dB HL (PTA of 500, 1000, 3000 Hz), PB word score of 1000 genes at p < 0.01) for most compounds. After ADME gene transcript enrichment, agglomerative clustering separated AhR ligands from CAR/PXR ligands, but it was difficult to distinguish CAR from PXR ligands. Consensus signatures were derived from groups of AhR, CAR and PXR ligands; and cross-talk among responding genes was determined. Many compounds had distinct log dose-response profiles, and relative potencies for ligands were established. Robust responses by CYP1A1, CYP2B10 (CAR responsive in mice) and CYP2B15 (CAR responsive in rats) and CYP3A1 (PXR responsive in rats) were used to benchmark the relative potency of different ligands and to determine the relative selectivity for AhR, CAR or PXR. By using a compendium of gene expression profiles, we defined species-specific induction patterns across the ADME transcriptome. PMID- 17118915 TI - Compendium of gene expression profiles comprising a baseline model of the human liver drug metabolism transcriptome. AB - Oligonucleotide microarrays were used to study the variability of pharmacokinetics and drug metabolism (PKDM)-related gene expression in 75 normal human livers. The objective was to define and use absorption, distribution, metabolism and excretion (ADME) gene expression variability to discern co regulated genes and potential surrogate biomarkers of inducible gene expression. RNA was prepared from donor tissue and hybridized on Agilent microarrays against an RNA mass balanced pool from all donors. Clustering of PKDM gene sets revealed donors with distinct patterns of gene expression that grouped genes known to be regulated by the nuclear receptor, pregnane X-receptor (PXR). Fold range metrics and frequency distributions from the heterogeneous human population were used to define the variability of individual PKDM genes in the 75 human livers and were placed in context by comparing expression data with basal ADME gene expression variability in an inbred and diet/environment controlled population of 27 Rhesus livers. The most variable genes in the hepatic transcriptome were mainly related to drug metabolism, intermediary metabolism, inflammation and cell cycle control. Unique patterns of expression across 75 individuals of inducible ADME gene expression allowed their expression to be correlated with the expression of many other genes. Correlated genes for AhR, CAR and PXR responsive genes (CYP1A2, CYP2B6 and CYP3A4) were identified that may be co-regulated and, therefore, provide clues to the identity of surrogate gene or protein markers for CYP induction. In conclusion, microarrays were used to define the variable expression of hepatic ADME genes in a diverse human population, the expression variability of ADME genes was compared with the expression variability in an inbred population of Rhesus monkeys, and genes were defined that may be co-regulated with important inducible CYP genes. PMID- 17118916 TI - Expression profiles of 50 xenobiotic transporter genes in humans and pre-clinical species: a resource for investigations into drug disposition. AB - Carrier-mediated transporters play a critical role in xenobiotic disposition and transporter research is complicated by species differences and their selective tissue expression. The purpose of this study was to generate a comprehensive data set of xenobiotic transporter gene expression profiles in humans and the pre clinical species mouse, rat, beagle dog and cynomolgus monkey. mRNA expression profiles of 50 genes from the ABC, SLC and SLCO transporter superfamilies were examined in 40 human tissues by microarray analyses. Transporter genes that were identified as enriched in the liver or kidney, or that were selected for their known roles in xenobiotic disposition, were then compared in 22 tissues across the five species. Finally, as clinical variability in drug response and adverse reactions may be the result of variability in transporter gene expression, variability in the expression of selected transporter genes in 75 human liver donors were examined and compared with the highly variable drug metabolizing enzyme CYP3A4. PMID- 17118917 TI - ADME transcriptome in Hispanic versus White donor livers: evidence of a globally enhanced NR1I3 (CAR, constitutive androstane receptor) gene signature in Hispanics. AB - Previous studies have found that, compared with Whites, Hispanic donor livers had elevated expression of CYP2 enzymes, gene products regulated by the constitutive androstane receptor (CAR). The objectives of the current study were to determine (1) the CAR activation signature in human liver (2) whether other drug detoxification (absorption, distribution, metabolism and excretion (ADME)) genes were differentially expressed in Hispanic versus White livers, and (3) the extent of overlap in the CAR and Hispanic liver transcriptomes. The CAR transcriptome (ADME genes differentially expressed following phenobarbital versus vehicle treatment of human hepatocytes) and the Hispanic liver transcriptome (ADME genes differentially expressed in Hispanic versus White livers) were identified using Affymetrix oligonucleotide arrays. Quantitative real-time polymerase chain reaction (PCR) was used to verify candidate genes in a larger sample size. Comparison of the CAR and Hispanic liver ADME transcriptomes revealed a significant association between the gene changes. Sixty-four per cent of the ADME genes induced more than twofold by phenobarbital were also induced in Hispanics, and 14% of the ADME genes repressed more than twofold by phenobarbital were repressed in Hispanics. In conclusion, compared with Whites, Hispanic donor livers have increased expression of many genes that are transcriptionally regulated by CAR. This result has practical implications to the drug treatment of Hispanic patients. PMID- 17118918 TI - Drug-induced changes in P450 enzyme expression at the gene expression level: a new dimension to the analysis of drug-drug interactions. AB - Drug-drug interactions (DDIs) caused by direct chemical inhibition of key drug metabolizing cytochrome P450 enzymes by a co-administered drug have been well documented and well understood. However, many other well-documented DDIs cannot be so readily explained. Recent investigations into drug and other xenobiotic mediated expression changes of P450 genes have broadened our understanding of drug metabolism and DDI. In order to gain additional information on DDI, we have integrated existing information on drugs that are substrates, inhibitors, or inducers of important drug-metabolizing P450s with new data on drug-mediated expression changes of the same set of cytochrome P450s from a large-scale microarray gene expression database of drug-treated rat tissues. Existing information on substrates and inhibitors has been updated and reorganized into drug-cytochrome P450 matrices in order to facilitate comparative analysis of new information on inducers and suppressors. When examined at the gene expression level, a total of 119 currently marketed drugs from 265 examined were found to be cytochrome P450 inducers, and 83 were found to be suppressors. The value of this new information is illustrated with a more detailed examination of the DDI between PPARalpha agonists and HMG-CoA reductase inhibitors. This paper proposes that the well-documented, but poorly understood, increase in incidence of rhabdomyolysis when a PPARalpha agonist is co-administered with a HMG-CoA reductase inhibitor is at least in part the result of PPARalpha-induced general suppression of drug metabolism enzymes in liver. The authors believe this type of information will provide insights to other poorly understood DDI questions and stimulate further laboratory and clinical investigations on xenobiotic-mediated induction and suppression of drug metabolism. PMID- 17118919 TI - Tissue and species distribution of the glutathione pathway transcriptome. AB - The goal of this study was to compare and contrast the basal gene expression levels of the various enzymes involved in glutathione metabolism among tissues and genders of the rat, mouse and canine. The approach taken was to use Affymetrix GeneChip microarray data for rat, mouse and canine tissues, comparing intensity levels for individual probes between tissues and genders. As was hypothesized, the relative expression in liver, lung, heart, kidney and testis varied from gene to gene, with differences of expression between tissues sometimes greater than a 1000-fold. The pattern of differential expression was usually similar between male and female animals, but varied greatly between the three species. Gstp1 appears to be expressed at high levels in male mouse liver, reasonable levels in canine liver, but very low levels in male rat liver. In all species examined, Gstp1 expression was below detectable levels in testis. Gsta3/Yc2 expression appeared high in rodent liver and female canine liver, but not male canine liver. Finally, Mgst1 and Gpx3 expression appeared to be lower in canine heart and testis than seen in rodents. Given the critical role of the glutathione pathway in the detoxification of many drugs and xenobiotics, the observed differences in basal tissue distribution among mouse, rat and canine has far-reaching implications in comparing responses of these species in safety testing. PMID- 17118920 TI - Predictive models of hepatotoxicity using gene expression data from primary rat hepatocytes. AB - With the aim of evaluating the usefulness of an in vitro system for assessing the potential hepatotoxicity of compounds, the paper describes several methods of obtaining mathematical models for the prediction of compound-induced toxicity in vivo. These models are based on data derived from treating rat primary hepatocytes with various compounds, and thereafter using microarrays to obtain gene expression 'profiles' for each compound. Predictive models were constructed so as to reduce the number of 'probesets' (genes) required, and subjected to rigorous cross-validation. Since there are a number of possible approaches to derive predictive models, several distinct modelling strategies were applied to the same data set, and the outcomes were compared and contrasted. While all the strategies tested showed significant predictive capability, it was interesting to note that the different approaches generated models based on widely disparate probesets. This implies that while these models may be useful in ascribing relative potential toxicity to compounds, they are unlikely to provide significant information on underlying toxicity mechanisms. Improved predictivity will be obtained through the generation of more comprehensive gene expression databases, covering more 'toxicity space', and by the development of models that maximize the observation, and combination, of individual differences between compounds. PMID- 17118921 TI - Emerging evidence for the interrelationship of xenobiotic exposure and circadian rhythms: a review. AB - The circadian clock controls many aspects of mammalian physiology and behaviour with a periodicity of approximately 24 h. These include the anticipation of, and adaptation to, daily environmental changes such as the light-dark cycle, temperature fluctuations and the availability of food. The toxicity of many drugs is dependent on the circadian phase at which they are administered, and recent work has begun to unravel the molecular basis for circadian variations in sensitivity to xenobiotic exposure. Between 2 and 10% of the transcriptome is expressed in a circadian manner, including many key genes associated with the metabolism and transport of xenobiotics. Furthermore, a number of xenobiotics may directly alter the expression of genes that control circadian rhythms. This review discusses the emerging evidence for the regulation of circadian rhythm genes having an important impact on molecular response to xenobiotics. PMID- 17118922 TI - Nuclear receptors CAR and PXR in the regulation of hepatic metabolism. AB - The nuclear receptors CAR and PXR were first characterized as xenosensing transcription factors regulating the induction of phase I and II xenobiotic metabolizing enzymes as well as transporters in response to exogenous stimuli. It has now become clear, however, that these receptors cross-talk with endogenous stimuli as well, which extends their regulation to various physiological processes such as energy metabolism and cell growth. As recognition of the function of these receptors has widened, the molecular mechanism of their regulation has evolved from simple protein-DNA binding to regulation by complex protein-protein interactions. Novel mechanisms as to how xenobiotic exposure alters hepatic metabolic pathways such as gluconeogenesis and beta-oxidation have emerged. At the same time, the molecular mechanism of how endogenous stimuli, such as insulin, regulate xenobiotc metabolism via CAR and PXR have also become evident. PMID- 17118923 TI - Introduction. The neurobiology of social recognition, attraction and bonding. PMID- 17118925 TI - Social odours, sexual arousal and pairbonding in primates. AB - We describe the role of social odours in sexual arousal and maintaining pairbonds in biparental and cooperatively breeding primates. Social odours are complex chemical mixtures produced by an organism that can simultaneously provide information about species, kinship, sex, individuality and reproductive state. They are long lasting and have advantages over other modalities. Both sexes are sensitive to changes in odours over the reproductive cycle and experimental disruption of signals can lead to altered sexual behaviour within a pair. We demonstrate, using functional magnetic resonance imaging (fMRI), that social odours indicating reproductive state directly influence the brain areas responsible for sexual behaviour. Social odours also influence other brain areas typically involved in motivation, memory and decision making, suggesting that these signals have more complex functions in primates than mere sexual arousal. We demonstrate a rapid link between social odours and neuroendocrine responses that are modulated by a male's social status. Recent work on humans shows similar responses to social odours. We conclude with an integration of the importance of social odours on sexual arousal and maintaining pairbonds in socially biparental and cooperatively breeding species, suggesting new research directions to integrate social behaviour, neural activation and neuroendocrine responses. PMID- 17118924 TI - Mammalian social odours: attraction and individual recognition. AB - Mammalian social systems rely on signals passed between individuals conveying information including sex, reproductive status, individual identity, ownership, competitive ability and health status. Many of these signals take the form of complex mixtures of molecules sensed by chemosensory systems and have important influences on a variety of behaviours that are vital for reproductive success, such as parent-offspring attachment, mate choice and territorial marking. This article aims to review the nature of these chemosensory cues and the neural pathways mediating their physiological and behavioural effects. Despite the complexities of mammalian societies, there are instances where single molecules can act as classical pheromones attracting interest and approach behaviour. Chemosignals with relatively high volatility can be used to signal at a distance and are sensed by the main olfactory system. Most mammals also possess a vomeronasal system, which is specialized to detect relatively non-volatile chemosensory cues following direct contact. Single attractant molecules are sensed by highly specific receptors using a labelled line pathway. These act alongside more complex mixtures of signals that are required to signal individual identity. There are multiple sources of such individuality chemosignals, based on the highly polymorphic genes of the major histocompatibility complex (MHC) or lipocalins such as the mouse major urinary proteins. The individual profile of volatile components that make up an individual odour signature can be sensed by the main olfactory system, as the pattern of activity across an array of broadly tuned receptor types. In addition, the vomeronasal system can respond highly selectively to non-volatile peptide ligands associated with the MHC, acting at the V2r class of vomeronasal receptor. The ability to recognize individuals or their genetic relatedness plays an important role in mammalian social behaviour. Thus robust systems for olfactory learning and recognition of chemosensory individuality have evolved, often associated with major life events, such as mating, parturition or neonatal development. These forms of learning share common features, such as increased noradrenaline evoked by somatosensory stimulation, which results in neural changes at the level of the olfactory bulb. In the main olfactory bulb, these changes are likely to refine the pattern of activity in response to the learned odour, enhancing its discrimination from those of similar odours. In the accessory olfactory bulb, memory formation is hypothesized to involve a selective inhibition, which disrupts the transmission of the learned chemosignal from the mating male. Information from the main olfactory and vomeronasal systems is integrated at the level of the corticomedial amygdala, which forms the most important pathway by which social odours mediate their behavioural and physiological effects. Recent evidence suggests that this region may also play an important role in the learning and recognition of social chemosignals. PMID- 17118926 TI - Voice processing in human and non-human primates. AB - Humans share with non-human primates a number of voice perception abilities of crucial importance in social interactions, such as the ability to identify a conspecific individual from its vocalizations. Speech perception is likely to have evolved in our ancestors on the basis of pre-existing neural mechanisms involved in extracting behaviourally relevant information from conspecific vocalizations (CVs). Studying the neural bases of voice perception in primates thus not only has the potential to shed light on cerebral mechanisms that may be- unlike those involved in speech perception--directly homologous between species, but also has direct implications for our understanding of how speech appeared in humans. In this comparative review, we focus on behavioural and neurobiological evidence relative to two issues central to voice perception in human and non human primates: (i) are CVs 'special', i.e. are they analysed using dedicated cerebral mechanisms not used for other sound categories, and (ii) to what extent and using what neural mechanisms do primates identify conspecific individuals from their vocalizations? PMID- 17118927 TI - The fusiform face area: a cortical region specialized for the perception of faces. AB - Faces are among the most important visual stimuli we perceive, informing us not only about a person's identity, but also about their mood, sex, age and direction of gaze. The ability to extract this information within a fraction of a second of viewing a face is important for normal social interactions and has probably played a critical role in the survival of our primate ancestors. Considerable evidence from behavioural, neuropsychological and neurophysiological investigations supports the hypothesis that humans have specialized cognitive and neural mechanisms dedicated to the perception of faces (the face-specificity hypothesis). Here, we review the literature on a region of the human brain that appears to play a key role in face perception, known as the fusiform face area (FFA). Section 1 outlines the theoretical background for much of this work. The face-specificity hypothesis falls squarely on one side of a longstanding debate in the fields of cognitive science and cognitive neuroscience concerning the extent to which the mind/brain is composed of: (i) special-purpose ('domain specific') mechanisms, each dedicated to processing a specific kind of information (e.g. faces, according to the face-specificity hypothesis), versus (ii) general-purpose ('domain-general') mechanisms, each capable of operating on any kind of information. Face perception has long served both as one of the prime candidates of a domain-specific process and as a key target for attack by proponents of domain-general theories of brain and mind. Section 2 briefly reviews the prior literature on face perception from behaviour and neurophysiology. This work supports the face-specificity hypothesis and argues against its domain-general alternatives (the individuation hypothesis, the expertise hypothesis and others). Section 3 outlines the more recent evidence on this debate from brain imaging, focusing particularly on the FFA. We review the evidence that the FFA is selectively engaged in face perception, by addressing (and rebutting) five of the most widely discussed alternatives to this hypothesis. In section 4, we consider recent findings that are beginning to provide clues into the computations conducted in the FFA and the nature of the representations the FFA extracts from faces. We argue that the FFA is engaged both in detecting faces and in extracting the necessary perceptual information to recognize them, and that the properties of the FFA mirror previously identified behavioural signatures of face-specific processing (e.g. the face-inversion effect). Section 5 asks how the computations and representations in the FFA differ from those occurring in other nearby regions of cortex that respond strongly to faces and objects. The evidence indicates clear functional dissociations between these regions, demonstrating that the FFA shows not only functional specificity but also area specificity. We end by speculating in section 6 on some of the broader questions raised by current research on the FFA, including the developmental origins of this region and the question of whether faces are unique versus whether similarly specialized mechanisms also exist for other domains of high-level perception and cognition. PMID- 17118928 TI - Genetic influences on the neural basis of social cognition. AB - The neural basis of social cognition has been the subject of intensive research in both human and non-human primates. Exciting, provocative and yet consistent findings are emerging. A major focus of interest is the role of efferent and afferent connectivity between the amygdala and the neocortical brain regions, now believed to be critical for the processing of social and emotional perceptions. One possible component is a subcortical neural pathway, which permits rapid and preconscious processing of potentially threatening stimuli, and it leads from the retina to the superior colliculus, to the pulvinar nucleus of the thalamus and then to the amygdala. This pathway is activated by direct eye contact, one of many classes of potential threat, and may be particularly responsive to the 'whites of the eyes'. In humans, autonomic arousal evoked by this stimulus is associated with the activity in specific cortical regions concerned with processing visual information from faces. The integrated functioning of these pathways is modulated by one or more X-linked genes, yet to be identified. The emotional responsiveness of the amygdala, and its associated circuits, to social threat is also influenced by functional polymorphisms in the promoter of the serotonin transporter gene. We still do not have a clear account of how specific allelic variation, in candidate genes, increases susceptibility to developmental disorders, such as autism, or psychiatric conditions, such as anxiety or depressive illness. However, the regulation of emotional responsiveness to social cues lies at the heart of the problem, and recent research indicates that we may be nearing a deeper and more comprehensive understanding. PMID- 17118929 TI - Reproductive strategy, sexual development and attraction to facial characteristics. AB - Sexual reproduction strategies vary both between and within species in the level of investment in offspring. Life-history theories suggest that the rate of sexual maturation is critically linked to reproductive strategy, with high investment being associated with few offspring and delayed maturation. For humans, age of puberty and age of first sex are two developmental milestones that have been associated with reproductive strategies. Stress during early development can retard or accelerate sexual maturation and reproduction. Early age of menarche is associated with absence of younger siblings, absence of a father figure during early life and increased weight. Father absence during early life is also associated with early marriage, pregnancy and divorce. Choice of partner characteristics is critical to successful implementation of sexual strategies. It has been suggested that sexually dimorphic traits (including those evident in the face) signal high-quality immune function and reproductive status. Masculinity in males has also been associated with low investment in mate and offspring. Thus, women's reproductive strategy should be matched to the probability of male investment, hence to male masculinity. Our review leads us to predict associations between the rate of sexual maturation and adult preferences for facial characteristics (enhanced sexual dimorphism and attractiveness). We find for men, engaging in sex at an early age is related to an increased preference for feminized female faces. Similarly, for women, the earlier the age of first sex the greater the preference for masculinity in opposite-sex faces. When we controlled sexual dimorphism in male faces, the speed of sexual development in women was not associated with differences in preference for male facial attractiveness. These developmental influences on partner choice were not mediated by self-rated attractiveness or parental relationships. We conclude that individuals assort in preferences based on the rapidity of their sexual development. Fast developing individuals prefer opposite-sex partners with an increased level of sexually dimorphic facial characteristics. PMID- 17118930 TI - Behavioural and neurophysiological evidence for face identity and face emotion processing in animals. AB - Visual cues from faces provide important social information relating to individual identity, sexual attraction and emotional state. Behavioural and neurophysiological studies on both monkeys and sheep have shown that specialized skills and neural systems for processing these complex cues to guide behaviour have evolved in a number of mammals and are not present exclusively in humans. Indeed, there are remarkable similarities in the ways that faces are processed by the brain in humans and other mammalian species. While human studies with brain imaging and gross neurophysiological recording approaches have revealed global aspects of the face-processing network, they cannot investigate how information is encoded by specific neural networks. Single neuron electrophysiological recording approaches in both monkeys and sheep have, however, provided some insights into the neural encoding principles involved and, particularly, the presence of a remarkable degree of high-level encoding even at the level of a specific face. Recent developments that allow simultaneous recordings to be made from many hundreds of individual neurons are also beginning to reveal evidence for global aspects of a population-based code. This review will summarize what we have learned so far from these animal-based studies about the way the mammalian brain processes the faces and the emotions they can communicate, as well as associated capacities such as how identity and emotion cues are dissociated and how face imagery might be generated. It will also try to highlight what questions and advances in knowledge still challenge us in order to provide a complete understanding of just how brain networks perform this complex and important social recognition task. PMID- 17118931 TI - Romantic love: a mammalian brain system for mate choice. AB - Mammals and birds regularly express mate preferences and make mate choices. Data on mate choice among mammals suggest that this behavioural 'attraction system' is associated with dopaminergic reward pathways in the brain. It has been proposed that intense romantic love, a human cross-cultural universal, is a developed form of this attraction system. To begin to determine the neural mechanisms associated with romantic attraction in humans, we used functional magnetic resonance imaging (fMRI) to study 17 people who were intensely 'in love'. Activation specific to the beloved occurred in the brainstem right ventral tegmental area and right postero-dorsal body of the caudate nucleus. These and other results suggest that dopaminergic reward and motivation pathways contribute to aspects of romantic love. We also used fMRI to study 15 men and women who had just been rejected in love. Preliminary analysis showed activity specific to the beloved in related regions of the reward system associated with monetary gambling for uncertain large gains and losses, and in regions of the lateral orbitofrontal cortex associated with theory of mind, obsessive/compulsive behaviours and controlling anger. These data contribute to our view that romantic love is one of the three primary brain systems that evolved in avian and mammalian species to direct reproduction. The sex drive evolved to motivate individuals to seek a range of mating partners; attraction evolved to motivate individuals to prefer and pursue specific partners; and attachment evolved to motivate individuals to remain together long enough to complete species-specific parenting duties. These three behavioural repertoires appear to be based on brain systems that are largely distinct yet interrelated, and they interact in specific ways to orchestrate reproduction, using both hormones and monoamines. Romantic attraction in humans and its antecedent in other mammalian species play a primary role: this neural mechanism motivates individuals to focus their courtship energy on specific others, thereby conserving valuable time and metabolic energy, and facilitating mate choice. PMID- 17118932 TI - Oxytocin, vasopressin and pair bonding: implications for autism. AB - Understanding the neurobiological substrates regulating normal social behaviours may provide valuable insights in human behaviour, including developmental disorders such as autism that are characterized by pervasive deficits in social behaviour. Here, we review the literature which suggests that the neuropeptides oxytocin and vasopressin play critical roles in modulating social behaviours, with a focus on their role in the regulation of social bonding in monogamous rodents. Oxytocin and vasopressin contribute to a wide variety of social behaviours, including social recognition, communication, parental care, territorial aggression and social bonding. The effects of these two neuropeptides are species-specific and depend on species-specific receptor distributions in the brain. Comparative studies in voles with divergent social structures have revealed some of the neural and genetic mechanisms of social-bonding behaviour. Prairie voles are socially monogamous; males and females form long-term pair bonds, establish a nest site and rear their offspring together. In contrast, montane and meadow voles do not form a bond with a mate and only the females take part in rearing the young. Species differences in the density of receptors for oxytocin and vasopressin in ventral forebrain reward circuitry differentially reinforce social-bonding behaviour in the two species. High levels of oxytocin receptor (OTR) in the nucleus accumbens and high levels of vasopressin 1a receptor (V1aR) in the ventral pallidum contribute to monogamous social structure in the prairie vole. While little is known about the genetic factors contributing to species-differences in OTR distribution, the species-specific distribution pattern of the V1aR is determined in part by a species-specific repetitive element, or 'microsatellite', in the 5' regulatory region of the gene encoding V1aR (avpr1a). This microsatellite is highly expanded in the prairie vole (as well as the monogamous pine vole) compared to a very short version in the promiscuous montane and meadow voles. These species differences in microsatellite sequence are sufficient to change gene expression in cell culture. Within the prairie vole species, intraspecific variation in the microsatellite also modulates gene expression in vitro as well as receptor distribution patterns in vivo and influences the probability of social approach and bonding behaviour. Similar genetic variation in the human AVPR1A may contribute to variations in human social behaviour, including extremes outside the normal range of behaviour and those found in autism spectrum disorders. In sum, comparative studies in pair bonding rodents have revealed neural and genetic mechanisms contributing to social-bonding behaviour. These studies have generated testable hypotheses regarding the motivational systems and underlying molecular neurobiology involved in social engagement and social bond formation that may have important implications for the core social deficits characterizing autism spectrum disorders. PMID- 17118933 TI - Mother-infant bonding and the evolution of mammalian social relationships. AB - A wide variety of maternal, social and sexual bonding strategies have been described across mammalian species, including humans. Many of the neural and hormonal mechanisms that underpin the formation and maintenance of these bonds demonstrate a considerable degree of evolutionary conservation across a representative range of these species. However, there is also a considerable degree of diversity in both the way these mechanisms are activated and in the behavioural responses that result. In the majority of small-brained mammals (including rodents), the formation of a maternal or partner preference bond requires individual recognition by olfactory cues, activation of neural mechanisms concerned with social reward by these cues and gender-specific hormonal priming for behavioural output. With the evolutionary increase of neocortex seen in monkeys and apes, there has been a corresponding increase in the complexity of social relationships and bonding strategies together with a significant redundancy in hormonal priming for motivated behaviour. Olfactory recognition and olfactory inputs to areas of the brain concerned with social reward are downregulated and recognition is based on integration of multimodal sensory cues requiring an expanded neocortex, particularly the association cortex. This emancipation from olfactory and hormonal determinants of bonding has been succeeded by the increased importance of social learning that is necessitated by living in a complex social world and, especially in humans, a world that is dominated by cultural inheritance. PMID- 17118935 TI - Genomic imprinting and the social brain. AB - Genomic imprinting refers to the parent-of-origin-specific epigenetic marking of a number of genes. This epigenetic mark leads to a bias in expression between maternally and paternally inherited imprinted genes, that in some cases results in monoallelic expression from one parental allele. Genomic imprinting is often thought to have evolved as a consequence of the intragenomic conflict between the parental alleles that occurs whenever there is an asymmetry of relatedness. The two main examples of asymmetry of relatedness are when there is partiality of parental investment in offspring (as is the case for placental mammals, where there is also the possibility of extended postnatal care by one parent), and in social groups where there is a sex-biased dispersal. From this evolutionary starting point, it is predicted that, at the behavioural level, imprinted genes will influence what can broadly be termed bonding and social behaviour. We examine the animal and human literature for examples of imprinted genes mediating these behaviours, and divide them into two general classes. Firstly, mother offspring interactions (suckling, attachment and maternal behaviours) that are predicted to occur when partiality in parental investment in early postnatal offspring occurs; and secondly, adult social interactions, when there is an asymmetry of relatedness in social groups. Finally, we return to the evolutionary theory and examine whether there is a pattern of behavioural functions mediated by imprinted genes emerging from the limited data, and also whether any tangible predictions can be made with regards to the direction of action of genes of maternal or paternal origin. PMID- 17118936 TI - Identification of mouse Prp19p as a lipid droplet-associated protein and its possible involvement in the biogenesis of lipid droplets. AB - Prp19p is an integral component of the heteromeric protein complex (the NineTeen complex) in the nucleus, and it is essential for the structural integrity of NineTeen complex and its subsequent activation of the spliceosome. We identified Prp19p, which has never been reported in relation to any function outside of the nucleus, as a member of proteins associated with lipid droplets. Down-regulation of Prp19p expression with RNA interference in 3T3-L1 cells repressed lipid droplet formation with the reduction in the level of expression of perilipin and S3-12. The levels of expression of SCD1 (stearoyl-CoA desaturase-1), DGAT-1 (acyl CoA diacylglycerol acyltransferase-1), and glycerol-3-phosphate acyltransferase were also reduced in Prp19p down-regulated cells, and a significant decrease in triglycerides was observed. Unlike perilipin, which is one of the most extensively studied lipid droplet-associated proteins, Prp19p is not essential for cAMP- and hormone-sensitive lipase-dependent lipolysis pathways, even though Prp19p is a component of the lipid droplet phospholipid monolayer, and down regulation of Prp19p represses fat accretion significantly. These results suggest that Prp19p or Prp19-interacting proteins during lipid droplet biogenesis in adipocytes may be considered as another class of potential targets for attacking obesity and obesity-related problems. PMID- 17118934 TI - Social buffering: relief from stress and anxiety. AB - Communication is essential to members of a society not only for the expression of personal information, but also for the protection from environmental threats. Highly social mammals have a distinct characteristic: when conspecific animals are together, they show a better recovery from experiences of distress. This phenomenon, termed 'social buffering', has been found in rodents, birds, non human primates and also in humans. This paper reviews classical findings on social buffering and focuses, in particular, on social buffering effects in relation to neuroendocrine stress responses. The social cues that transmit social buffering signals, the neural mechanisms of social buffering and a partner's efficacy with respect to social buffering are also detailed. Social contact appears to have a very positive influence on the psychological and the physiological aspects of social animals, including human beings. Research leading towards further understanding of the mechanisms of social buffering could provide alternative medical treatments based on the natural, individual characteristics of social animals, which could improve the quality of life. PMID- 17118937 TI - In vitro activity of Citrus bergamia (bergamot) oil against clinical isolates of dermatophytes. AB - OBJECTIVES: Recently, bergamot oil was shown to be a potent antifungal agent in vitro against clinically important Candida species. In this study, the activities of bergamot natural essence and its furocoumarin-free and distilled extracts on dermatophytes such as Trichophyton, Microsporum and Epidermophyton species were investigated. METHODS: In vitro susceptibility testing assays on 92 clinical isolates of dermatophytes (Trichophyton mentagrophytes n = 20, Trichophyton rubrum n = 18, Trichophyton interdigitale n = 15, Trichophyton tonsurans n = 2, Microsporum canis n = 24, Microsporum gypseum n = 1 and Epidermophyton floccosum n = 12) were performed using the CLSI M38-A broth microdilution method, except for employing an inoculum of 1-3 x 10(3) cfu/mL. MICs were determined at a visual endpoint reading of 80% inhibition compared with the growth control. RESULTS: MICs (v/v) of all fungi ranged from 0.156% to 2.5% for the natural essence, from 0.02% to 2.5% for the distilled extract, and from 0.08% to 1.25% for the furocoumarin-free extract. The three isolates of T. tonsurans and M. gypseum exhibited the highest MIC values. CONCLUSIONS: Data from this study indicate that bergamot oil is active in vitro against several common species of dermatophytes, suggesting its potential use for topical treatment of dermatophytoses. PMID- 17118938 TI - Antibiotic-resistant Campylobacter: could efflux pump inhibitors control infection? AB - Campylobacter is the most common cause of bacterial gastroenteritis in the world. Poultry is the main reservoir of human infections. The widespread use of antibiotics in agriculture and veterinary medicine has resulted in the emergence of an increasing number of antibiotic-resistant Campylobacter strains that can be transmitted to humans through the food chain. Of particular concern to public health is the prevalence of resistance to macrolides and fluoroquinolones that are used in the treatment of life-threatening campylobacteriosis. The CmeABC efflux system has been shown to contribute to the intrinsic and acquired resistance to these antibiotics. In addition, by mediating resistance to bile, it is essential for colonization of the chicken gut in vivo. Inhibition of CmeABC may provide an effective means of reversing antibiotic resistance and decreasing the transmission of Campylobacter via the food chain. This would positively impact on public health by decreasing the morbidity, mortality and increased healthcare costs associated with the treatment of antibiotic-resistant Campylobacter. PMID- 17118939 TI - Susceptibility of Actinobacillus actinomycetemcomitans to six antibiotics decreases as biofilm matures. AB - OBJECTIVES: Actinobacillus actinomycetemcomitans is a major causative agent of chronic and aggressive periodontitis. Freshly isolated strains of A. actinomycetemcomitans display rough-type colonies and initiate biofilm formation on glass surfaces. The purpose of this study was to determine the antibiotic susceptibility of A. actinomycetemcomitans biofilm during different phases of maturation. METHODS: Using 96-well microtitre plates, we determined the antibiotic susceptibility of rough-type strain 310a to concentrations from 0.1 to 10 mg/L each of erythromycin, ofloxacin, ampicillin, cefalexin, tetracycline and minocycline during biofilm formation. Antibiotics were added at the start of the culture (early phase) and after 24 h of cultivation (mature phase). RESULTS: Adding 10 mg/L of ampicillin, 10 mg/L of cefalexin, 0.1 or 1 mg/L of tetracycline, or 0.1 mg/L of minocycline significantly inhibited 310a biofilm formation in the early phase, but not in the mature phase. Although adding 10 mg/L of erythromycin, tetracycline or minocycline reduced biofilm development in the early phase, it enhanced 310a biofilm development in the mature phase. Ofloxacin exerted a strong inhibitory effect in both the early and mature phases of biofilm formation throughout all experiments. CONCLUSIONS: The present study demonstrated that the susceptibility of A. actinomycetemcomitans to many antibiotics decreased after biofilm maturation. PMID- 17118940 TI - Chronic shoulder pain and diaphragmatic endometriosis. PMID- 17118941 TI - Wegener's granulomatosis presenting as a disappearing renal mass. PMID- 17118943 TI - Successful treatment of primitive neuroectodermal tumor-associated microangiopathy with multiple bone metastases. AB - We report here a 16-year-old male with primitive neuroectodermal tumor (PNET) associated probable microangiopathy with multiple bone metastases. Laboratory findings excluded the possibility of amegakaryocytic or immune thrombocytopenia and/or disseminated intravascular coagulation. He was first treated with plasma exchange (PE), followed by platelet transfusions, steroid pulse therapy and combined chemotherapy. PE and steroid pulse therapy reduced his plasma CRP level. Combined chemotherapy drastically increased his platelet count until it had almost normalized without further transfusion. The plasma level of von Willebrand factor-cleaving protease (ADAMTS13) activity measured before PE was not severely deficient (48% of normal) and an unusually large von Willebrand factor multimer (UL-VWFM) was detected. We consider that this therapeutic strategy has the following benefits: (1) reduction of plasma levels of factors that are harmful to both platelet activation and endothelial cell injury; and (2) the safe transfusion of platelet concentrate in thrombotic microangiopathy. This strategy should be confirmed in further cases. PMID- 17118944 TI - Status report on Medicare Part D enrollment in 2006: analysis of plan-specific market share and coverage. AB - The centerpiece of the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 was the Part D drug benefit, provided through new stand-alone prescription drug plans (PDPs) and Medicare Advantage prescription drug (MA-PD) plans. We examine 2006 Part D enrollment data to analyze organization- and plan level market share and enrollment by plan type, benefit design, and gap coverage. Ten organizations captured 72 percent of Part D enrollment, primarily in low premium plans and those with name recognition. More than twelve million Part D enrollees without low-income subsidies enrolled in plans with limited or no gap coverage in 2006, but the number with actual spending in the gap remains to be seen. PMID- 17118945 TI - Application of wavelet-based tools to study the dynamics of biological processes. AB - The article makes use of three different examples (sensory information processing in the rat trigeminal complex, intracellular interaction in snail neurons and multimodal dynamics in nephron autoregulation) to demonstrate how modern approaches to time-series analysis based on the wavelet-transform can provide information about the underlying complex biological processes. PMID- 17118946 TI - Heat shock-induced changes in lipid and protein metabolism in the endoplasmic reticulum of barley aleurone layers. AB - Heat shock in barley aleurone layers induces heat shock protein synthesis and suppresses secretory protein synthesis by selectively destabilizing their mRNAs. In addition, the endoplasmic reticulum (ER) membranes upon which secretory protein mRNAs are translated become vesiculated during heat shock, leading to the hypothesis that ER dissociation and targeted mRNA destabilization are linked mechanistically. Supporting this, ER can be heat adapted, and heat-adapted ER has higher levels of fatty acid saturation in membrane phospholipids which do not vesiculate upon heat shock. Secretory protein mRNAs are also more stable in heat adapted cells. To understand better heat shock-induced changes in ER membranes, we examined ER membrane proteins and enzymes involved in phosphatidylcholine biosynthesis and phospholipid turnover in heat-shocked aleurone cells. Heat shock significantly increased the activity of phospholipases A2 and D, and shortly thereafter significant but gradual increases in choline kinase and phosphocholine glyceride transferase activities and a sharp increase in phosphorylcholine citidyl transferase activity were observed. Only minor changes were observed in SDS-PAGE analyses of proteins from sonicated ER membranes fractionated on continuous sucrose gradients. Overall, heat shock reduced total lipid in ER membranes relative to protein, and in intact, ultracentrifuged aleurone cells examined by light and electron microscopy the ER band appeared to increase in density. The changes in phospholipid metabolism coupled with the suppression of secretory protein synthesis indicate that in addition to inducing a classic heat shock response, high temperature also induces a classic unfolded protein response in the ER of this secretory cell. PMID- 17118947 TI - Disruption of actin filaments by latrunculin B affects cell wall construction in Picea meyeri pollen tube by disturbing vesicle trafficking. AB - The involvement of actin filaments (AFs) in vesicle trafficking, cell wall construction and tip growth was investigated during pollen tube development of Picea meyeri. Pollen germination and tube elongation were inhibited in a dose dependent manner by the latrunculin B (LatB) treatment. The fine AFs were broken down into disorganized fragments showing a tendency to aggregate. FM4-64 labeling revealed that the dynamic balance of vesicle trafficking was perturbed due to F actin disruption and the fountain-like cytoplasmic pattern changed into disorganized Brownian movement. The configuration and/or distribution of cell wall components, such as pectins, callose and cellulose, as well as arabinogalactan proteins changed in obvious ways after the LatB application. Fourier transform infrared (FTIR) analysis further established significant changes in the chemical composition of the wall material. Our results indicate that depolymerization of AFs affects the distribution and configuration of cell wall components in Picea meyeri pollen tube by disturbing vesicle trafficking. PMID- 17118948 TI - Utilization and transport of mannitol in Olea europaea and implications for salt stress tolerance. AB - Mannitol is one of the primary photosynthetic products and the major phloem translocated carbohydrate in Olea europaea L., an important crop in the Mediterranean basin. Uptake of mannitol in heterotrophic cell suspensions of O. europaea was shown to be mediated by a 1 : 1 polyol : H+ symport system with a Km of 1.3 mM mannitol and a Vmax of 1.3 nmol min(-1) mg(-1) DW. Dulcitol, sorbitol and xylitol competed for mannitol uptake, whereas glucose and sucrose did not. Reverse transcription-PCR (RT-PCR) performed on mRNA extracted from cultured cells exhibiting high mannitol transport activity allowed the cloning of a partial O. europaea mannitol carrier OeMaT1. The Vmax of mannitol uptake and the amount of OeMaT1 transcripts increased along with polyol depletion from the medium, suggesting that the mannitol transport system may be regulated by its own substrate. Addition of 100-500 mM NaCl to cultured cells enhanced the capacity of the polyol : H+ symport system and the amount of OeMaT1 transcripts, whereas it strongly repressed mannitol dehydrogenase activity. Measurements of cell viability showed that mannitol-grown cells remained viable 24 h after a 250 and 500 mM NaCl pulse, whereas extensive loss of cell viability was observed in sucrose-grown cells. OeMaT1 transcripts increased throughout maturation of olive fruits, suggesting that an OeMaT is involved in the accumulation of mannitol during ripening of olive. Thus, mannitol transport and compartmentation by OeMaT are important to allocate this source of carbon and energy, as well as for salt tolerance and olive ripening. PMID- 17118949 TI - Self management for men with lower urinary tract symptoms: randomised controlled trial. AB - OBJECTIVE: To evaluate the effectiveness of self management as a first line intervention for men with lower urinary tract symptoms. DESIGN: Randomised controlled trial. SETTING: A teaching hospital and a district general hospital in London. PARTICIPANTS: 140 men (mean age 63 (SD 10.7) years), recruited between January 2003 and April 2004, referred by general practitioners to urological outpatient departments with uncomplicated lower urinary tract symptoms. INTERVENTIONS: Self management and standard care (n=73) or standard care alone (n=67). The self management group took part in three small group sessions comprising education, lifestyle advice, and training in problem solving and goal setting skills. MAIN OUTCOME MEASURES: The primary outcome measure was treatment failure measured at 3, 6, and 12 months. Symptom severity (international prostate symptom score; higher scores represent a poorer outcome) was used as a secondary outcome. RESULTS: At three months, treatment failure had occurred in 7 (10%) of the self management group and in 27 (42%) of the standard care group (difference=32%, 95% confidence interval 18% to 46%). Corresponding differences in the frequency of treatment failure were 42% (27% to 57%) at six months and 48% (32% to 64%) at 12 months. At three months, the mean international prostate symptom score was 10.7 in the self management group and 16.4 in the standard care group (difference=5.7, 3.7 to 7.7). Corresponding differences in score were 6.5 (4.3 to 8.7) at six months and 5.1 (2.7 to 7.6) at 12 months. CONCLUSIONS: Self management significantly reduced the frequency of treatment failure and reduced urinary symptoms. Because of the large observed benefit of self management, the results of this study support the case for a large multicentre trial to confirm whether self management could be considered as first line treatment for men with lower urinary tract symptoms. TRIAL REGISTRATION: National Research Register N0263115137; Clinical trials NCT00270309 [ClinicalTrials.gov]. PMID- 17118950 TI - Impact of a theoretically based sex education programme (SHARE) delivered by teachers on NHS registered conceptions and terminations: final results of cluster randomised trial. AB - OBJECTIVE: To assess the impact of a theoretically based sex education programme (SHARE) delivered by teachers compared with conventional education in terms of conceptions and terminations registered by the NHS. DESIGN: Follow-up of cluster randomised trial 4.5 years after intervention. SETTING: NHS records of women who had attended 25 secondary schools in east Scotland. PARTICIPANTS: 4196 women (99.5% of those eligible). INTERVENTION: SHARE programme (intervention group) v existing sex education (control group). MAIN OUTCOME MEASURE: NHS recorded conceptions and terminations for the achieved sample linked at age 20. RESULTS: In an "intention to treat" analysis there were no significant differences between the groups in registered conceptions per 1000 pupils (300 SHARE v 274 control; difference 26, 95% confidence interval -33 to 86) and terminations per 1000 pupils (127 v 112; difference 15, -13 to 42) between ages 16 and 20. CONCLUSIONS: This specially designed sex education programme did not reduce conceptions or terminations by age 20 compared with conventional provision. The lack of effect was not due to quality of delivery. Enhancing teacher led school sex education beyond conventional provision in eastern Scotland is unlikely to reduce terminations in teenagers. TRIAL REGISTRATION: ISRCTN48719575 [controlled trials.com]. PMID- 17118951 TI - Structures of unique globoside elongation products present in erythrocytes with a rare NOR phenotype. AB - Rare polyagglutinable erythrocytes of NOR phenotype were found to contain two unique glycosphingolipids (designated NOR1 and NOR2). These components (not detected in normal erythrocytes) were reactive with Griffonia simplicifolia isolectin IB4 (GSL-IB4) and commonly present human anti-NOR antibodies. The NOR1 component has been reported to be a globoside containing a single galactose residue linked alpha1,4 to the terminal N-acetylgalactosamine. Here, we report the structural studies on a second glycolipid, NOR2, and a third novel component migrating in high-performance thin-layer chromatography (HPTLC) between NOR1 and NOR2. The structures were determined by a combination of ion trap sequential mass spectrometry (MALDI-QIT-TOF) and step-wise treatment with glycosidases, followed by identification of products on HPTLC plates with lectins and mouse monoclonal anti-NOR antibody. The NOR2 component was found to be a disaccharide extension of NOR1 with the following structure: Galalpha1-4GalNAcbeta1-3Galalpha1-4GalNAcbeta1 3Galalpha1-4Galbeta1-4Glcbeta1-Cer. Treatment of NOR2 with alpha-galactosidase gave a glycolipid migrating between NOR1 and NOR2, which did not react with either GSL-IB4 or anti-NOR antibodies but did react with GalNAc-specific soybean agglutinin. This intermediate glycolipid (now designated NOR(int)) was identified as a relatively abundant component of a neutral glycolipid fraction from NOR erythrocytes, suggesting its presence as a precursor to NOR2. The structure of NOR(int) was also confirmed by sequential mass spectrometry studies. These results indicate that polyagglutination in NOR subjects is due to unique erythrocyte glycolipids that are synthesized by sequential addition of Galalpha1,4 and GalNAcbeta1,3 to globoside. PMID- 17118952 TI - Communicable disease and health protection quarterly review: April to June 2006: from the Health Protection Agency. PMID- 17118953 TI - A multi-centre evaluation of nine rapid, point-of-care syphilis tests using archived sera. AB - OBJECTIVES: To evaluate nine rapid syphilis tests at eight geographically diverse laboratory sites for their performance and operational characteristics. METHODS: Tests were compared "head to head" using locally assembled panels of 100 archived (50 positive and 50 negative) sera at each site using as reference standards the Treponema pallidum haemagglutination or the T pallidum particle agglutination test. In addition inter-site variation, result stability, test reproducibility and test operational characteristics were assessed. RESULTS: All nine tests gave good performance relative to the reference standard with sensitivities ranging from 84.5-97.7% and specificities from 84.5-98%. Result stability was variable if result reading was delayed past the recommended period. All the tests were found to be easy to use, especially the lateral flow tests. CONCLUSIONS: All the tests evaluated have acceptable performance characteristics and could make an impact on the control of syphilis. Tests that can use whole blood and do not require refrigeration were selected for further evaluation in field settings. PMID- 17118954 TI - Diagnosis of gonococcal infection in high risk women using a rapid test. AB - OBJECTIVE: To assess the performance and acceptability for patients and health care workers of the NGThermo Biostar (GC OIA) to diagnose gonococcal infection compared with culture using modified Thayer Martin medium. METHODS: This study involved 326 high-risk women presenting with vaginal discharge or referral by sexual partner with urethral discharge at a sexually transmitted infections (STI) clinic in Manaus, Brazil. Endocervical swabs collected from the women were tested with both the NG Biostar and modified Thayer Martin culture as the reference standard test. Clinic staff were trained to perform the NG Biostar on site and the culture was performed in the laboratory of the clinic. RESULTS: The prevalence of gonococcal infection as measured by the reference standard was 15% (50/326) overall. Among asymptomatic participants, the prevalence of infection was 17.7% (25/141) and among symptomatic women it was 13.5% (25/185) (p = 0.3). Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for the NG Biostar test, with 95% confidence intervals (CI), were 60% (46.4% to 73.6%), 89.9% (86.2% to 93.6%), 55.6% (42.4% to 68.8%), and 92.6% (89.5% to 95.7%), respectively; 98.8% of study participants were willing to wait approximately 1 hour in the clinic for test results. CONCLUSION: Syndromic management protocols for treatment of STI in developing countries require refinement because, as currently described, they lead to over-treatment of cervical infection. A rapid test done during patients' initial presentation and leading to immediate treatment if positive would help improve the accuracy of diagnosis and could also be used to screen asymptomatic women. Even though the NG Biostar had a low sensitivity and PPV, which is less than ideal, it could still improve the rates of treatment over the gold standard test that requires return visits for patients to receive results and to benefit from treatment. Cost effectiveness studies using rapid point-of-care tests for Neisseria gonorrhoeae infection compared to the syndromic approach should be carried out to assess their value in STI diagnosis and treatment in developing nations. PMID- 17118955 TI - Plasma pro-B-type natriuretic peptide in the general population: screening for left ventricular hypertrophy and systolic dysfunction. AB - AIMS: B-type natriuretic peptide (BNP) measurement in screening for left ventricular hypertrophy (LVH) and left ventricular systolic dysfunction (LVSD) has been evaluated in the general population, but corresponding information on proBNP and the N-terminal proBNP fragment is still limited. We therefore examined whether proBNP measurement is useful for LVH and LVSD screening in the general population. METHODS AND RESULTS: In the 4th Copenhagen City Heart Study, 3497 participants underwent echocardiography with assessment of left ventricular ejection fraction (LVEF) and mass. The impact of gender and age was determined and the diagnostic performance of the plasma proBNP concentration was evaluated using receiver operating characteristic (ROC) curves. Of 1502 men and 1995 women, 4.1 and 2.6% had LVSD defined as an LVEF<60% whereas only 0.4% displayed LVEF<40%. The proBNP concentration was 1.7-fold higher in women compared with men (P<0.0001) and related to age in both genders. The mean proBNP plasma concentration was two-fold higher in subjects with LVSD than without LVSD (P<0.0001). Likewise, LVH imposed a 1.9-fold increase in the proBNP concentration (P<0.0001): Both differences persisted after adjusting for ischaemic heart disease, hypertension, diabetes, gender, and age. The diagnostic performance of proBNP in detecting LVEF<40% was high with an area under the ROC curves of 0.92 (95% CI 0.79-1.00) in women and 0.85 (95% CI 0.74-0.96) in men. CONCLUSION: We have established the impact of age and gender on the proBNP concentration in a large, community-based cohort. The diagnostic performance for proBNP measurement in screening for LVH and LVSD in the general population parallels the reported data for BNP. PMID- 17118956 TI - NvMap: automated analysis of NMR chemical shift perturbation data. AB - NMR chemical shift perturbation experiments are widely used to define binding sites in biomolecular complexes. Especially in the case of high throughput screening of ligands, rapid analysis of NMR spectra is essential. NvMap extends NMRViewJ and provides a means for rapid assignments and book-keeping of NMR titration spectra. Our module offers options to analyze multiple titration spectra both separately and sequentially, where the sequential spectra are analyzed either two at a time or all simultaneously. The first option is suitable for slow or intermediate exchange rates between free and bound proteins. The latter option is particularly useful for fast exchange situations and can compensate for the lack of indicators for overlapped peaks. Our module also provides a simple user interface to automate the analysis process from dataset to peak list. We demonstrate the effectiveness of our program using NMR spectra of SUMO in complexes with three different peptides. AVAILABILITY: NvMap is available on the web at http://www.cityofhope.org/Researchers/ChenYuan/NvMap/ Supplemental information: Manual pages and test spectra will be available on the web at the above site. PMID- 17118957 TI - A framework for gene expression analysis. AB - MOTIVATION: Global gene expression measurements as obtained, for example, in microarray experiments can provide important clues to the underlying transcriptional control mechanisms and network structure of a biological cell. In the absence of a detailed understanding of this gene regulation, current attempts at classification of expression data rely on clustering and pattern recognition techniques employing ad-hoc similarity criteria. To improve this situation, a better understanding of the expected relationships between expression profiles of genes associated by biological function is required. RESULTS: It is shown that perturbation expansions familiar from biological systems theory make precise predictions for the types of relationships to be expected for expression profiles of biologically associated genes, even if the underlying biological factors responsible for this association are not known. Classification criteria are derived, most of which are not usually employed in clustering algorithms. The approach is illustrated by using the AtGenExpress Arabidopsis thaliana developmental expression map. PMID- 17118958 TI - PartTree: an algorithm to build an approximate tree from a large number of unaligned sequences. AB - MOTIVATION: To construct a multiple sequence alignment (MSA) of a large number (> approximately 10,000) of sequences, the calculation of a guide tree with a complexity of O(N2) to O(N3), where N is the number of sequences, is the most time-consuming process. RESULTS: To overcome this limitation, we have developed an approximate algorithm, PartTree, to construct a guide tree with an average time complexity of O(N log N). The new MSA method with the PartTree algorithm can align approximately 60,000 sequences in several minutes on a standard desktop computer. The loss of accuracy in MSA caused by this approximation was estimated to be several percent in benchmark tests using Pfam. AVAILABILITY: The present algorithm has been implemented in the MAFFT sequence alignment package (http://align.bmr.kyushu-u.ac.jp/mafft/software/). SUPPLEMENTARY INFORMATION: Supplementary information is available at Bioinformatics online. PMID- 17118959 TI - WHAP: haplotype-based association analysis. AB - We describe a software tool to perform haplotype-based association analysis, for quantitative and qualitative traits, in population and family samples, using single nucleotide polymorphism or multiallelic marker data. A range of tests is offered: omnibus and haplotype-specific tests; prospective and retrospective likelihoods; covariates and moderators; sliding window analyses; permutation P values. We focus on the ability to flexibly impose constraints on haplotype effects, which allows for a range of conditional haplotype-based likelihood ratio tests: for example, whether an allele has an effect independent of its haplotypic background, or whether a single variant can explain the overall association at a locus. We illustrate using these tests to dissect a multi-locus association. AVAILABILITY: WHAP is a C/C++ program, freely available from the author's website: http://pngu.mgh.harvard.edu/purcell/whap/ PMID- 17118960 TI - Azaspiracid-1 alters the E-cadherin pool in epithelial cells. AB - Azaspiracids cause severe damages in the epithelium of several organs. In this study we have investigated the effects of azaspiracid-1 (AZA-1) on two epithelial cell lines. Nanomolar concentrations of AZA-1 reduced MCF-7 cell proliferation and impaired cell-cell adhesion. AZA-1 altered the cellular pool of the adhesion molecule E-cadherin by inducing a dose- and time-dependent accumulation of an E cadherin fragment (E-cadherin-related antigen [ECRA(100)]), with a concentration inducing the half-maximal effect (EC(50)) of 0.47nM. The immunological characterization of ECRA(100) revealed that it consists of an E-cadherin molecule lacking the intracellular domain, and these data showed that the effect induced by AZA-1 in MCF-7 cells is undistinguishable from that induced by yessotoxin (YTX) in the same experimental system. A comparison of toxin effects in MCF-7 and Caco 2 cells confirmed that the effects induced by AZA-1 and YTX are undistinguishable in these cells. Treatment of fibroblasts with AZA-1 did not affect the cellular pool of N-cadherin showing that the toxin effect is cadherin specific. A comparison of the effects induced by AZA-1, YTX, and okadaic acid on F-actin and E-cadherin in MCF-7 and Caco 2 cells showed that 1nM AZA-1 did not cause significant changes in F-actin and that accumulation of ECRA(100) did not correlate with decreased levels of F-actin under our experimental conditions. Matching our results with those available in literature, we notice that, when molecular effects induced by AZA-1 and YTX have been studied in the same in vitro systems, experimental data show that they are undistinguishable in terms of sensitive cellular parameters, effective doses, and kinetics of responses in several cell lines. The possibility that azaspiracids and YTXs might share their molecular mechanism(s) of action in defined biological settings should be considered. PMID- 17118961 TI - An unexpected role for keratin 10 end domains in susceptibility to skin cancer. AB - Keratin 10 (K10) is a type I keratin that is expressed in post-mitotic suprabasal keratinocytes of the skin. Based on cell culture experiments and transgenic mouse studies, it has been proposed that K10 suppresses cell proliferation and tumor formation in the skin. Furthermore, the ability of K10 to suppress cell proliferation was mapped to its unique N- and C-terminal protein domains. In the present study, we modified the endogenous keratin 14 (K14) gene of mice using a knock-in approach to encode a chimeric keratin that consists of the K14 rod domain fused to the K10 head and tail domains (K1014chim). This transgene was expressed in the basal layer of the epidermis and the outer root sheath of hair follicles. Unexpectedly, we found that the K10 end domains had no effect on basal keratinocyte proliferation in vivo. Moreover, when subjected to a chemical skin carcinogenesis protocol, papilloma formation in mutant mice was accelerated instead of being inhibited. Our data suggest that the increased tumor susceptibility of K1014chim mice is in part due to a suppression of apoptosis in mutant keratinocytes. Our results support the notion that intermediate filaments, in addition to their function as cytoskeletal components, affect tumor susceptibility of epithelial cells. PMID- 17118962 TI - The Cdi/TESK1 kinase is required for Sevenless signaling and epithelial organization in the Drosophila eye. AB - How cellular behaviors such as cell-to-cell communication, epithelial organization and cell shape reorganization are coordinated during development is poorly understood. The developing Drosophila eye offers an ideal model system to study these processes. Localized actin polymerization is required to constrict the apical surface of epithelial cells of the eye imaginal disc to maintain the refined arrangement of the developing ommatidia. The identity of each photoreceptor cell within the epithelium is determined by cell-to-cell contacts involving signal transduction events. The R7 photoreceptor cell requires the activity of the Sevenless RTK to adopt a proper cell fate. We performed an EP screen for negative regulators of this inductive process, and we identified the serine/threonine kinase Center divider (cdi) as a suppressor of the phenotype caused by an activated Sevenless receptor. Cdi is homologous to the human testis specific kinase 1 (TESK1), a member of the LIM kinases involved in cytoskeleton control through ADF/cofilin phosphorylation. We have analyzed the effects of gain and loss-of-function of cdi and found alterations in actin organization and in the adherens junctions proteins DE-cadherin and beta-catenin, as well as in Sevenless apical localization. Interference with the function of the ADF/cofilin phosphatase Slingshot (ssh), which antagonizes Cdi, also results in a suppression of signaling triggered by the Sevenless RTK. Our results reveal a critical interplay between the localization of molecules involved in epithelial organization and signal transduction. PMID- 17118963 TI - Werner syndrome protein participates in a complex with RAD51, RAD54, RAD54B and ATR in response to ICL-induced replication arrest. AB - Werner syndrome (WS) is a rare genetic disorder characterized by genomic instability caused by defects in the WRN gene encoding a member of the human RecQ helicase family. RecQ helicases are involved in several DNA metabolic pathways including homologous recombination (HR) processes during repair of stalled replication forks. Following introduction of interstrand DNA crosslinks (ICL), WRN relocated from nucleoli to arrested replication forks in the nucleoplasm where it interacted with the HR protein RAD52. In this study, we use fluorescence resonance energy transfer (FRET) and immune-precipitation experiments to demonstrate that WRN participates in a multiprotein complex including RAD51, RAD54, RAD54B and ATR in cells where replication has been arrested by ICL. We verify the WRN-RAD51 and WRN-RAD54B direct interaction in vitro. Our data support a role for WRN also in the recombination step of ICL repair. PMID- 17118964 TI - PDZRN3 (LNX3, SEMCAP3) is required for the differentiation of C2C12 myoblasts into myotubes. AB - PDZRN3 contains a RING-finger motif in its N-terminal region, two PDZ domains in its central region and a consensus-binding motif for PDZ domains at its C terminus. It was identified in silico as a homolog of the protein known as LNX1 or SEMCAP1, which possesses ubiquitin ligase activity and binds the membrane protein Semaphorin 4C. However, PDZRN3 itself has not previously been characterized. We have now evaluated the properties and functions of PDZRN3. The PDZRN3 gene was shown to be expressed in various human tissues including the heart, skeletal muscle and liver and its expression in mouse skeletal muscle was developmentally regulated. Both the differentiation of C2C12 mouse skeletal myoblasts into myotubes and injury-induced muscle regeneration in vivo were found to be accompanied by up-regulation of PDZRN3. The differentiation-associated increase in the expression of PDZRN3 in C2C12 cells follows that of myogenin and precedes that of myosin heavy chain. Depletion of PDZRN3 by RNA interference inhibited the formation of myotubes as well as the associated up-regulation of myosin heavy chain in C2C12 cells. Our data suggest that PDZRN3 plays an essential role in the differentiation of myoblasts into myotubes by acting either downstream or independently of myogenin. PMID- 17118965 TI - Dietary carbohydrate, glycemic index, and glycemic load in relation to risk of colorectal cancer in women. AB - Diets with a high glycemic index and glycemic load have been hypothesized to be implicated in the etiology of colorectal cancer owing to their potential to increase postprandial glucose and insulin levels. Prospective data on glycemic index and glycemic load in relation to colorectal cancer risk are limited and inconsistent. Therefore, the authors prospectively investigated the associations of dietary carbohydrate, glycemic index, and glycemic load with the incidence of colorectal cancer among 61,433 Swedish women who were free of cancer in 1987-1990 and completed a 67-item food frequency questionnaire. During follow-up through June 2005, 870 incident cases of colorectal adenocarcinoma were diagnosed. Carbohydrate intake, glycemic index, and glycemic load were not associated with risk of colorectal cancer, colon cancer, or rectal cancer. The multivariate hazard ratios for colorectal cancer comparing the highest with the lowest quintile were 1.10 (95% confidence interval: 0.85, 1.44) for carbohydrate intake, 1.00 (95% confidence interval: 0.75, 1.33) for glycemic index, and 1.06 (95% confidence interval: 0.81, 1.39) for glycemic load. Results did not vary by body mass index. The findings from this prospective study do not support the hypothesis that a high carbohydrate intake, a high glycemic index, and a high glycemic load increase the risk of colorectal cancer. PMID- 17118966 TI - Increased antigen presentation and T(h)1 polarization in genetically histamine free mice. AB - Histamine is a well-known inflammatory mediator exerting various immunomodulatory effects and affecting the development of antigen-specific immune responses. Dendritic cells (DCs) are the most potent antigen-presenting cells specialized for capture, uptake, transport, processing and presentation of antigens to T cells. Using a genetically histamine-free [histidine decarboxylase knockout (HDC /-)] mouse model, we examined the effects of histamine on DC-mediated antigen presentation. Applying an in vitro antigen presentation assay, we found that spleen DCs, derived from HDC-/- mice, display a higher efficiency in antigen presentation compared with wild-type cells. Flow cytometric characterization of DCs disclosed that this difference was not due to an altered distribution of DCs between or within the major functional sub-populations (assessed by CD11b and CD4 as myeloid and CD8alpha and DEC205 as lymphoid DC markers) or major changes in the co-stimulatory molecule profile (CD40, CD80, CD86). However, real-time PCR analysis of in vivo CFA-induced IL-12p35, IFNgamma, IL-10 and IL-4 expression showed that DCs matured in a histamine-free environment exhibit significantly elevated levels of IL-12p35 and IFNgamma mRNA. In vitro investigations confirmed that isolated DCs, developed in the absence of histamine, exhibit indeed a predominantly T(h)1-polarized cytokine pattern, as they show elevated levels of IFNgamma mRNA upon LPS stimulation. Similar difference was found at the protein level by ELISA, as well. Our study demonstrates that histamine interferes with antigen presentation and alters the cytokine profile of DCs. PMID- 17118967 TI - WSX-1 plays a significant role for the initiation of experimental autoimmune uveitis. AB - WSX-1 is a subunit of the IL-27R, which plays a critical role in the initiation of T(h)1 responses. Murine experimental autoimmune uveitis (EAU) is a model of human autoimmune uveitis, in which a T(h)1 response predominates in the pathogenetic process. To explore the role of WSX-1 in this model, WSX-1(-/-) mice were immunized with interphotoreceptor retinoid-binding protein peptide 1-20 to induce EAU. We found that the EAU clinical and histological scores were lower in the WSX-1(-/-) mice up to day 21, whereas after day 21, the EAU scores were the same between the wild-type (WT) and WSX-1(-/-) mice with both declining at the same rate. In contrast to T lymphocytes from WT mice, WSX-1(-/-) T lymphocytes on day 9 after immunization failed to produce IFN-gamma. Similarly, expression of T(h)1-related chemokines, such as regulated on activation, normal T cell expressed and secreted and IP-10, in the eye was reduced in WSX-1(-/-) mice compared with WT mice on day 13 after immunization. In addition, sub-retinal transfer of lymphocytes from WSX-1(-/-) mice on day 9 after immunization did not induce EAU in the recipient mice. Importantly, IFN-gamma production, chemokine expression and the transferability of disease by lymphocytes became comparable for WSX-1(-/-) and WT mice at later stages. Thus, IL-27/WSX-1 affects the early development of EAU, and might be a target for therapy during the onset of autoimmune uveitis in humans. PMID- 17118968 TI - 172G>T variant in the 5' untranslated region of DNA repair gene RAD51 reduces risk of squamous cell carcinoma of the head and neck and interacts with a P53 codon 72 variant. AB - RAD51 participates in homologous recombination (HR) repair of double-stranded DNA breaks (DSBs) that may cause genomic instability and cancer. Two single nucleotide polymorphisms (SNPs) and three P53 binding sites have been found in the RAD51 promoter and 5' untranslated region. We hypothesized that RAD51 and P53 SNPs may interact and alter risk of squamous cell carcinoma of the head and neck (SCCHN) and we genotyped for RAD51 135G>C and 172G>T and P53 Arg72Pro SNPs in 716 SCCHN patients and 719 matched controls (all non-Hispanic whites) and evaluated their effects on gamma radiation-induced mutagen sensitivity. We found that RAD51 172TT homozygotes had a significantly decreased risk [adjusted odds ratio (OR) = 0.66, 95% confidence interval (CI) = 0.50-0.87] of SCCHN, compared with carriers of other genotypes, particularly in P53 Arg72Arg homozygotes (adjusted OR = 0.60, 95% CI = 0.41-0.89) (homogeneity test P = 0.047), although no alterations in the risk were associated with the RAD51 135G>C and P53 Arg72Pro SNPs. Consistent with a protective effect of the 172TT genotype, significantly fewer gamma radiation induced chromatid breaks per cell were present in 172TT homozygotes (mean +/- SD = 0.36 +/- 0.13) than in subjects with other genotypes (mean +/- SD = 0.46 +/- 0.13, P < 0.001) among 148 control subjects we tested. The finding that the functional RAD51 172G>T SNP, particularly in the presence of the P53 Arg72Arg genotype, may be a marker of susceptibility to SCCHN needs to be validated by larger studies of different ethnic populations. PMID- 17118969 TI - Relationships between IQ and regional cortical gray matter thickness in healthy adults. AB - Prior studies show positive correlations between full-scale intelligence quotient (FSIQ) and cerebral gray matter measures. Few imaging studies have addressed whether general intelligence is related to regional variations in brain tissue and the associated influences of sex. Cortical thickness may more closely reflect cytoarchitectural characteristics than gray matter density or volume estimates. To identify possible localized relationships, we examined FSIQ associations with cortical thickness at high spatial resolution across the cortex in healthy young adult (age 17-44 years) men (n = 30) and women (n = 35). Positive relationships were found between FSIQ and intracranial gray and white matter but not cerebrospinal fluid volumes. Significant associations with cortical thickness were evident bilaterally in prefrontal (Brodmann's areas [BAs] 10/11, 47) and posterior temporal cortices (BA 36/37) and proximal regions. Sex influenced regional relationships; women showed correlations in prefrontal and temporal association cortices, whereas men exhibited correlations primarily in temporal occipital association cortices. In healthy adults, greater intelligence is associated with larger intracranial gray matter and to a lesser extent with white matter. Variations in prefrontal and posterior temporal cortical thickness are particularly linked with intellectual ability. Sex moderates regional relationships that may index dimorphisms in cognitive abilities, overall processing strategies, or differences in structural organization. PMID- 17118970 TI - Multispectral fluorescence and reflectance imaging at the leaf level and its possible applications. AB - Images taken at different spectral bands are increasingly used for characterizing plants and their health status. In contrast to conventional point measurements, imaging detects the distribution and quantity of signals and thus improves the interpretation of fluorescence and reflectance signatures. In multispectral fluorescence and reflectance set-ups, images are separately acquired for the fluorescence in the blue, green, red, and far red, as well as for the reflectance in the green and in the near infrared regions. In addition, 'reference' colour images are taken with an RGB (red, green, blue) camera. Examples of imaging for the detection of photosynthetic activity, UV screening caused by UV-absorbing substances, fruit quality, leaf tissue structure, and disease symptoms are introduced. Subsequently, the different instrumentations used for multispectral fluorescence and reflectance imaging of leaves and fruits are discussed. Various types of irradiation and excitation light sources, detectors, and components for image acquisition and image processing are outlined. The acquired images (or image sequences) can be analysed either directly for each spectral range (wherein they were captured) or after calculating ratios of the different spectral bands. This analysis can be carried out for different regions of interest selected manually or (semi)-automatically. Fluorescence and reflectance imaging in different spectral bands represents a promising tool for non-destructive plant monitoring and a 'road' to a broad range of identification tasks. PMID- 17118971 TI - Suillus variegatus causes significant changes in the content of individual polyamines and flavonoids in Scots pine seedlings during mycorrhiza formation in vitro. AB - Changes in the concentrations of individual flavonoids and polyamines (PAs) in Scots pine (Pinus sylvestris L.) cotyledonary seedlings were studied during the establishment of an ectomycorrhizal (ECM) symbiosis with two Suillus variegatus strains in vitro. Both flavonoids and PAs were analysed after 3, 7, and 14 d in dual culture, and changes in concentrations were compared with growth of the seedlings. Both S. variegatus strains caused similar responses in Scots pine seedlings. Free putrescine accumulated immediately but only transiently after inoculation. This was followed by continuous accumulation of PA conjugates in needles and stems, and free spermidine and spermine in roots, which was accompanied by mycorrhiza formation and improved growth. The fungi induced lateral root formation and main root and primary needle elongation. Inoculation caused no qualitative changes in flavonoid composition, while quantitative changes in flavonols, catechins, and condensed tannins were observed in shoots during mycorrhiza formation. These results indicate that in this in vitro system conjugated PAs and specific flavonoids, generally related to the plant's defence reactions, did not play a major role in the regulation of the establishment of the ectomycorrhizal (ECM) symbiosis in Scots pine roots. The results also clearly show that positive growth responses in shoots and roots due to S. variegatus were supported by different and highly specific changes in the synthesis of both primary and secondary metabolites in these parts of the seedling. PMID- 17118972 TI - Time-course metabolic profiling in Arabidopsis thaliana cell cultures after salt stress treatment. AB - Salt stress is one of the most important factors limiting plant cultivation. Many investigations of plant response to high salinity have been performed using conventional transcriptomics and/or proteomics approaches. However, transcriptomics and proteomics techniques are not all-encompassing methods that can achieve exclusive insights into the metabolite networks contributing to biochemical reactions. Hence, the functions of the complex stress response pathways are yet to be determined, especially at the metabolic level. A time course metabolic profiling with Arabidopsis thaliana cell cultures after the imposition of salt stress is reported in this study. Analyses of primary metabolites, especially small polar metabolites such as amino acids, sugars, sugar alcohols, organic acids, and amines, was performed by GC/MS and LC/MS at 0.5, 1, 2, 4, 12, 24, 48, and 72 h after a salt-stress treatment with 100 mM NaCl being the final concentration. The mass chromatographic data were converted into matrix data sets, which were subjected to data mining processes, including principal component analysis (PCA) and batch-learning self-organizing mapping analysis (BL-SOM). The mining results suggest that the methylation cycle for the supply of methyl groups, the phenylpropanoid pathway for lignin production, and glycinebetaine biosynthesis are synergetically induced as a short-term response against salt-stress treatment. The results also suggest the the co-induction of glycolysis and sucrose metabolism as well as co-reduction of the methylation cycle as long-term responses to salt stress. PMID- 17118973 TI - Perceptual anomalies in schizophrenia: integrating phenomenology and cognitive neuroscience. AB - From phenomenological and experimental perspectives, research in schizophrenia has emphasized deficits in "higher" cognitive functions, including attention, executive function, as well as memory. In contrast, general consensus has viewed dysfunctions in basic perceptual processes to be relatively unimportant in the explanation of more complex aspects of the disorder, including changes in self experience and the development of symptoms such as delusions. We present evidence from phenomenology and cognitive neuroscience that changes in the perceptual field in schizophrenia may represent a core impairment. After introducing the phenomenological approach to perception (Husserl, the Gestalt School), we discuss the views of Paul Matussek, Klaus Conrad, Ludwig Binswanger, and Wolfgang Blankenburg on perception in schizophrenia. These 4 psychiatrists describe changes in perception and automatic processes that are related to the altered experience of self. The altered self-experience, in turn, may be responsible for the emergence of delusions. The phenomenological data are compatible with current research that conceptualizes dysfunctions in perceptual processing as a deficit in the ability to combine stimulus elements into coherent object representations. Relationships of deficits in perceptual organization to cognitive and social dysfunction as well as the possible neurobiological mechanisms are discussed. PMID- 17118974 TI - Enteropathogenic Escherichia coli, Shigella flexneri, and Listeria monocytogenes recruit a junctional protein, zonula occludens-1, to actin tails and pedestals. AB - Enteropathogenic Escherichia coli, Shigella flexneri, and Listeria monocytogenes induce localized actin polymerization at the cytoplasmic face of the plasma membrane or within the host cytoplasm, creating unique actin-rich structures termed pedestals or actin tails. The process is known to be mediated by the actin related protein 2 and 3 (Arp2/3) complex, which in these cases acts downstream of neural Wiskott-Aldrich syndrome protein (N-WASP) or of a listerial functional homolog of WASP family proteins. Here, we show that zonula occludens-1 (ZO-1), a protein in the tight junctions of polarized epithelial cells, is recruited to actin tails and pedestals. Immunocytochemical analysis revealed that ZO-1 was stained most in the distal part of the actin-rich structures, and the incorporation was mediated by the proline-rich region of the ZO-1 molecule. The direct clustering of membrane-targeted Nck, which is known to activate the N-WASP Arp2/3 pathway, triggered the formation of the ZO-1-associated actin tails. The results suggest that the activation of the Arp2/3 complex downstream of N-WASP or a WASP-related molecule is a key to the formation of the particular actin-rich structures that bind with ZO-1. We propose that an analysis of the recruitment on a molecular basis will lead to an understanding of how ZO-1 recognizes a distinctive actin-rich structure under pathophysiological conditions. PMID- 17118975 TI - A genomic island of the pathogen Leptospira interrogans serovar Lai can excise from its chromosome. AB - An examination of the two Leptospira interrogans genomes sequenced so far reveals few genetic differences, including an extra DNA region, 54 kb in length, in L. interrogans serovar Lai. This locus contains 103 predicted coding sequences that are absent from the genome of L. interrogans serovar Copenhageni, of which only 20% had significant BLASTP hits in GenBank. By analyzing the L. interrogans serovar Lai genome by pulsed-field gel electrophoresis, we also found that this 54-kb DNA fragment exists as a circular plasmid. This was confirmed by amplification of a DNA fragment corresponding to that of the predicted fragment if this region excised from the chromosome and its left and right ends joined together. In addition, cloning of the putative rep gene of this DNA region was responsible for autonomous replication in Leptospira spp., therefore generating a new Escherichia coli-Leptospira sp. shuttle vector. Taken together, our results show that this genomic island can excise from the chromosome and form a replicative plasmid. Analysis of the distribution of this genomic island revealed that highly related sequences exist in other L. interrogans virulent strains. This genomic island, containing a high proportion of novel genes, may have an important role in spreading genes, including virulence factors, among bacterial populations. PMID- 17118976 TI - Therapeutic Chlamydophila abortus and C. pecorum vaccination transiently reduces bovine mastitis associated with Chlamydophila infection. AB - Infections with Chlamydophila abortus and C. pecorum are highly prevalent in cattle and have been associated with bovine mastitis. A prospective cohort study was conducted with a herd of 140 Holstein dairy cows to investigate the influence of Chlamydophila infection on subclinical inflammation of the bovine mammary gland as characterized by somatic cell numbers in milk. PCR detection of C. abortus and low serum antibody levels against Chlamydophila spp. were significantly associated with subclinical mastitis. To examine the effect of the infection by response modification, immune perturbation was done by two subcutaneous administrations of an experimental vaccine preparation of inactivated C. abortus and C. pecorum elementary bodies. Vaccination against Chlamydophila highly significantly decreased milk somatic cell numbers, thus reducing bovine mastitis, and increased antibody levels against Chlamydophila but did not eliminate shedding of C. abortus in milk as detected by PCR. The protective effect peaked at 11 weeks after vaccination and lasted for a total of 14 weeks. Vaccination with the Chlamydophila vaccine, a mock vaccine, or a combination vaccine against bovine viral diseases highly significantly increased C. abortus shedding in milk for 1 week, presumably mediated by the vaccine adjuvant. In summary, this study shows an etiological involvement of the widespread Chlamydophila infections in bovine mastitis, a herd disease of critical importance for the dairy industry. Furthermore, this investigation shows the potential for temporary improvement of chlamydial disease by therapeutic vaccination. Chlamydophila vaccination of cattle might serve as a testing ground for vaccines against human chlamydial infections. PMID- 17118977 TI - Differential activation of human gingival epithelial cells and monocytes by Porphyromonas gingivalis fimbriae. AB - Humans develop periodontitis in response to challenge by microbial dental plaque. Inflammation begins after perturbation of gingival epithelial cells by subgingival bacteria interacting through pattern-recognition receptors, including the Toll-like receptors (TLR). Porphyromonas gingivalis is a major periodontopathogen that interacts with epithelial cells through its cell surface fimbriae (FimA), leading to colonization and/or invasion. Previous work by our group has established membrane CD14 as an essential coreceptor for TLR2-mediated activation of transfected cell lines by P. gingivalis FimA. We have shown that gingival epithelial cells express TLR2 but not CD14 on their cell surfaces. We thus speculated that P. gingivalis FimA does not readily activate epithelial innate immune responses but rather functions to promote P. gingivalis colonization in the absence of a vigorous FimA-induced response. This hypothesis was verified by the findings that primary human gingival epithelial cells responded poorly to FimA in terms of interleukin (IL)-6, IL-8, granulocyte macrophage colony-stimulating factor, and tumor necrosis factor alpha responses, in stark contrast to the marked response to other TLR2 agonists (Pam3Cys, FSL-1) that are not strictly dependent on CD14. On the other hand, CD14-expressing human primary monocytes responded with high levels of the same cytokines to both FimA and the control TLR2 agonists. The gingival epithelial cells failed to respond to FimA even in the presence of exogenously added soluble CD14. These data indicate that the gingival epithelial cell hyporesponsiveness to FimA is attributable to the lack of membrane-expressed but not soluble CD14. In conclusion, P. gingivalis FimA differentially activates human monocytes and epithelial cells, perhaps reflecting different tactics used by P. gingivalis when interacting with different host cell types or a host strategy to limit inflammation. PMID- 17118978 TI - Vaccination with live Yersinia pestis primes CD4 and CD8 T cells that synergistically protect against lethal pulmonary Y. pestis infection. AB - Vaccination with live attenuated Yersinia pestis confers protection against pneumonic plague but is not considered safe for general use. Subunit plague vaccines containing the Y. pestis F1 and LcrV proteins prime robust antibody responses but may not provide sufficient protection. To aid the development of a safe and effective plague vaccine, we are investigating roles for T cells during defense against Y. pestis infection. Here we demonstrate that vaccination of mice with live Y. pestis primes specific CD4 and CD8 T cells that, upon purification and direct transfer to naive mice, synergistically protect against lethal intranasal Y. pestis challenge. While not preventing extrapulmonary dissemination, the coadministered T cells promote bacterial clearance and reduce bacteremia. These observations strongly suggest that development of pneumonic plague vaccines should strive to prime both CD4 and CD8 T cells. Finally, we demonstrate that vaccination with live Y. pestis primes CD4 and CD8 T cells that respond to Y. pestis strains lacking the capacity to express F1, LcrV, and all pCD1/pPCP-encoded proteins, suggesting that protective T cells likely recognize antigens distinct from those previously defined as targets for humoral immunity. PMID- 17118979 TI - Contribution of phagocytosis and intracellular sensing for cytokine production by Staphylococcus aureus-activated macrophages. AB - Toll-like receptors (TLRs) are involved in the sensing of microbially derived compounds. We analyzed the contribution of these receptors to cytokine production by macrophages following stimulation with whole bacteria. Using knockout mice, we confirmed that the TLR4 and TLR2 contribution was predominant in the induction of tumor necrosis factor alpha and interleukin-10 by gram-negative bacteria. In contrast, the absence of TLR2 and/or TLR4 or TLR9 did not affect the response to gram-positive bacteria. In the absence of TLR2, phagocytosis was essential for cytokine production in response to heat-killed Staphylococcus aureus (HKSA). Because intracellular sensing was important in the absence of TLR2, we evaluated the contribution of Nod1 and Nod2, intracytoplasmic sensors of peptidoglycan derived muropeptides, to the response to HKSA. By transfecting RAW 264.7 macrophages with dominant negative (DN) forms of Nod1 and Nod2, we showed that both molecules inhibited NF-kappaB activation in response to HKSA. The unexpected interference of DN Nod1 in the response of macrophages to gram-positive bacteria was confirmed with a Nod2 agonist (muramyl dipeptide) in transfection experiments with HEK293T cell. Taken together, these results show the contribution of phagocytosis and Nod molecules to the response to HKSA in macrophages and also identify possible cross talk between Nod1 and Nod2. PMID- 17118980 TI - ActA is required for crossing of the fetoplacental barrier by Listeria monocytogenes. AB - The facultative intracellular bacterial pathogen Listeria monocytogenes induces severe fetal infection during pregnancy. Little is known about the molecular mechanisms allowing the maternofetal transmission of bacteria. In this work, we studied fetoplacental invasion by infecting mice with various mutants lacking virulence factors involved in the intracellular life cycle of L. monocytogenes. We found that the placenta was highly susceptible to bacteria, including avirulent bacteria, such as an L. monocytogenes mutant with an hly deletion (DeltaLLO) and a nonpathogenic species, Listeria innocua, suggesting that permissive trophoblastic cells, trapping bacteria, provide a protective niche for bacterial survival. The DeltaLLO mutant, which is unable to escape the phagosomal compartment of infected cells, failed to grow in the trophoblast tissue and to invade the fetus. Mutant bacteria with inlA and inlB deletion (DeltaInlAB) grew in the placenta and fetus as well as did the wild-type virulent stain (EGDwt), indicating that in the murine model, internalins A and B are not involved in fetoplacental invasion by L. monocytogenes. Pregnant mice were then infected with an actA deletion (DeltaActA) strain, a virulence-attenuated mutant that is unable to polymerize actin and to spread from cell to cell. With the DeltaActA mutant, fetal infection occurs, but with a significant delay and restriction, and it requires a placental bacterial load 2 log units higher than that for the wild type virulent strain. Definitive evidence for the role of ActA was provided by showing that a actA-complemented DeltaActA mutant was restored in its capacity to invade fetuses. ActA-mediated cell-to-cell spreading plays a major role in the vertical transmission of L. monocytogenes to the fetus in the murine model. PMID- 17118981 TI - How flagellin and toll-like receptor 5 contribute to enteric infection. PMID- 17118982 TI - Mutants of type II heat-labile enterotoxin LT-IIa with altered ganglioside binding activities and diminished toxicity are potent mucosal adjuvants. AB - The structure and function LT-IIa, a type II heat-labile enterotoxin of Escherichia coli, are closely related to the structures and functions of cholera toxin and LT-I, the type I heat-labile enterotoxins of Vibrio cholerae and enterotoxigenic Escherichia coli, respectively. While LT-IIa is a potent systemic and mucosal adjuvant, recent studies demonstrated that mutant LT-IIa(T34I), which exhibits no detectable binding activity as determined by an enzyme-linked immunosorbent assay, with gangliosides GD1b, GD1a, and GM1 is a very poor adjuvant. To evaluate whether other mutant LT-IIa enterotoxins that also exhibit diminished ganglioside-binding activities have greater adjuvant activities, BALB/c mice were immunized by the intranasal route with the surface adhesin protein AgI/II of Streptococcus mutans alone or in combination with LT-IIa, LT IIa(T14S), LT-IIa(T14I), or LT-IIa(T14D). All three mutant enterotoxins potentiated strong mucosal immune responses that were equivalent to the response promulgated by wt LT-IIa. All three mutant enterotoxins augmented the systemic immune responses that correlated with their ganglioside-binding activities. Only LT-IIa and LT-IIa(T14S), however, enhanced expression of major histocompatibility complex class II and the costimulatory molecules CD40, CD80, and CD86 on splenic dendritic cells. LT-IIa(T14I) and LT-IIa(T14D) had extremely diminished toxicities in a mouse Y1 adrenal cell bioassay and reduced abilities to induce the accumulation of intracellular cyclic AMP in a macrophage cell line. PMID- 17118984 TI - Growth condition-dependent Esp expression by Enterococcus faecium affects initial adherence and biofilm formation. AB - A genetic subpopulation of Enterococcus faecium, called clonal complex 17 (CC 17), is strongly associated with hospital outbreaks and invasive infections. Most CC-17 strains contain a putative pathogenicity island encoding the E. faecium variant of enterococcal surface protein (Esp). Western blotting, flow cytometric analyses, and electron microscopy showed that Esp is expressed and exposed on the surface of E. faecium, though Esp expression and surface exposure are highly varied among different strains. Furthermore, Esp expression depends on growth conditions like temperature and anaerobioses. When grown at 37 degrees C, five of six esp-positive E. faecium strains showed significantly increased levels of surface-exposed Esp compared to bacteria grown at 21 degrees C, which was confirmed at the transcriptional level by real-time PCR. In addition, a significant increase in surface-exposed Esp was found in half of these strains when grown at 37 degrees C under anaerobic conditions compared to the level in bacteria grown under aerobic conditions. Finally, amounts of surface-exposed Esp correlated with initial adherence to polystyrene (R(2) = 0.7146) and biofilm formation (R(2) = 0.7535). Polystyrene adherence was competitively inhibited by soluble recombinant N-terminal Esp. This study demonstrates that Esp expression on the surface of E. faecium (i) varies consistently between strains, (ii) is growth condition dependent, and (iii) is quantitatively correlated with initial adherence and biofilm formation. These data indicate that E. faecium senses and responds to changing environmental conditions, which might play a role in the early stages of infection when bacteria transit from oxygen-rich conditions at room temperature to anaerobic conditions at body temperature. In addition, variation of surface exposure may explain the contrasting findings reported on the role of Esp in biofilm formation. PMID- 17118983 TI - Adipocyte, adipose tissue, and infectious disease. PMID- 17118985 TI - Expression of phosphorylcholine by Histophilus somni induces bovine platelet aggregation. AB - Histophilus somni-induced platelet aggregation was inhibited by antagonists of the platelet-activating factor (PAF) receptor but not inhibitors of PAF synthesis. In addition, H. somni cells expressing phosphorylcholine (ChoP) induced aggregation, while ChoP(-) H. somni cells did not. This suggests that H. somni ChoP may induce platelet aggregation via interactions with the PAF receptor. PMID- 17118986 TI - Oxidant generation by single infected monocytes after short-term fluorescence labeling of a protozoan parasite. AB - Leishmania spp. are intracellular protozoa residing in mononuclear phagocytes. Leishmania organisms are susceptible to microbicidal responses generated in response to phagocytosis. Assuming that both phagocyte and parasite populations are heterogeneous, it is advantageous to examine the response of individual cells phagocytosing living parasites. Because Leishmania spp. lose virulence during the raising of transfectants, we developed a method to label live Leishmania chagasi short-term with fluorescent dyes. Up to six parasite divisions were detected by flow cytometry after labeling with carboxyfluorescein diacetate succinimidyl ester (CFSE), dioctadecyl-tetramethylindo carbocyanine perchlorate, or chloromethyl tetramethylrhodamine. Labeled parasites entered mononuclear phagocytes as determined by confocal and time-lapse microscopy. Dihydroethidium (DHE) was used to detect macrophage-derived oxidants generated during phagocytosis. Presumably Leishmania organisms are opsonized with host serum/tissue components such as complement prior to phagocytosis. Therefore, we investigated the effects of opsonization and found that this increased the efficiency of CFSE-labeled parasite entry into monocytes (84.6% +/- 8.8% versus 20.2% +/- 3.8% monocytes infected; P < 0.001). Opsonization also increased the percentage of phagocytes undergoing a respiratory burst (66.0% +/- 6.3% versus 41.0% +/- 8.3% of monocytes containing CFSE-labeled parasites; P < 0.001) and the magnitude of oxidant generation by each infected monocyte. Inhibitor data indicated that DHE was oxidized by products of the NADPH oxidase. These data suggest that opsonized serum components such as complement lead to more efficient entry of Leishmania into their target cells but at the same time activate the phagocyte oxidase to generate microbicidal products in infected cells. The parasite must balance these positive and negative survival effects in order to initiate a viable infection. PMID- 17118987 TI - Intranasal vaccination with a secreted chlamydial protein enhances resolution of genital Chlamydia muridarum infection, protects against oviduct pathology, and is highly dependent upon endogenous gamma interferon production. AB - There is currently no licensed vaccine against Chlamydia trachomatis, the leading cause of sexually transmitted bacterial disease worldwide. Conventional vaccination attempts using surface-exposed chlamydial antigens have achieved only partial success. We have employed a novel vaccination strategy using a secreted protein, chlamydial protease-like activity factor (CPAF), which has been shown to degrade host major histocompatibility complex transcription factors and keratin-8 and therefore may allow immune evasion and establishment of a productive infection. Intranasal immunization using recombinant CPAF (rCPAF) plus interleukin-12 (IL-12) (rCPAF+IL-12 immunization) was used to assess the protective immunity against genital Chlamydia muridarum infection in BALB/c mice. rCPAF+IL-12 immunization induced robust gamma interferon (IFN-gamma) production and minimal IL-4 production by splenocytes upon in vitro recall with rCPAF. The total and immunoglobulin G2a (IgG2a) anti-rCPAF antibody levels in serum were significantly elevated after rCPAF+IL-12 vaccination, as were the total antibody, IgG2a, and IgA levels in bronchoalveolar lavage and vaginal fluids when the animals were compared to animals that received rCPAF alone. rCPAF+IL-12 vaccinated mice displayed significantly reduced bacterial shedding upon chlamydial challenge and accelerated resolution of infection compared to mock immunized (phosphate-buffered saline) animals. Moreover, rCPAF+IL-12-immunized animals exhibited protection against pathological consequences of chlamydial infection, including the development of hydrosalpinx and oviduct dilatation. This vaccination regimen also reduced the development of fibrosis and the influx of neutrophils into the upper genital tract when the animals were compared to mock immunized (phosphate-buffered saline) animals after bacterial challenge. rCPAF+IL 12-mediated resolution of the bacterial infection and protection against Chlamydia-induced inflammatory disease were highly dependent on endogenous IFN gamma production. Together, these results demonstrate that secreted chlamydial antigens may be novel vaccine candidates to induce protective immunity. PMID- 17118988 TI - The human immunodeficiency virus type 1 tat protein enhances Cryptosporidium parvum-induced apoptosis in cholangiocytes via a Fas ligand-dependent mechanism. AB - While Cryptosporidium parvum infection of the intestine has been reported in both immunocompetent and immunocompromised individuals, biliary infection is seen primarily in adult AIDS patients and is associated with development of AIDS cholangiopathy. However, the mechanisms of pathogen-induced AIDS cholangiopathy remain unclear. Since we previously demonstrated that the Fas/Fas ligand (FasL) system is involved in paracrine-mediated C. parvum cytopathicity in cholangiocytes, we also tested the potential synergistic effects of human immunodeficiency virus type 1 (HIV-1) transactivator of transcription (Tat) mediated FasL regulation on C. parvum-induced apoptosis in cholangiocytes by semiquantitative reverse transcription-PCR, immunoblotting, immunofluorescence analysis, and immunogold electron microscopy. H69 cells do not express CXCR4 and CCR5, which are receptors required for direct HIV-1 viral infection. However, recombinant biologically active HIV-1-associated Tat protein increased FasL expression in the cytoplasm of cholangiocytes without a significant increase in apoptosis. We found that C. parvum-induced apoptosis was associated with translocation of intracellular FasL to the cell membrane surface and release of full-length FasL from infected H69 cells. Tat significantly (P < 0.05) increased C. parvum-induced apoptosis in bystander cells in a dose-dependent manner. Moreover, Tat enhanced both C. parvum-induced FasL membrane translocation and release of full-length FasL. In addition, the FasL neutralizing antibody NOK-1 and the caspase-8 inhibitor Z-IETD-fmk both blocked C. parvum-induced apoptosis in cholangiocytes. The data demonstrated that HIV-1 Tat enhances C. parvum induced cholangiocyte apoptosis via a paracrine-mediated, FasL-dependent mechanism. Our results suggest that concurrent active HIV replication, with associated production of Tat protein, and C. parvum infection synergistically increase cholangiocyte apoptosis and thus jointly contribute to AIDS-related cholangiopathies. PMID- 17118989 TI - Expression of merozoite surface protein markers by Plasmodium falciparum-infected erythrocytes in peripheral blood and tissues of children with fatal malaria. AB - Sequestration of Plasmodium falciparum-infected erythrocytes is a pathological feature of fatal cerebral malaria. P. falciparum is genetically diverse among, and often within, patients. Preferential sequestration of certain genotypes might be important in pathogenesis. We compared circulating parasites with parasites sequestered in the brain, spleen, liver, and lung in the same Malawian children with fatal malaria, classifying serotypes using antibodies to merozoite surface proteins 1 and 2 and immunofluorescence in order to differentiate parasites and to quantify the proportions of each serotype. We found (i) similar distributions of various serotypes in different tissues and (ii) concordance between parasite serotypes in peripheral blood and parasite serotypes in tissues. No serotypes predominated in the brain in cerebral malaria, and parasites belonging to a single serotype did not cluster within individual vessels or within single tissues. These findings do not support the hypothesis that cerebral malaria is caused by cerebral sequestration of certain virulent types. PMID- 17118990 TI - Autoimmune polyendocrine syndrome type 1 in Norway: phenotypic variation, autoantibodies, and novel mutations in the autoimmune regulator gene. AB - CONTEXT: The autoimmune polyendocrine syndrome type I (APS I) is a rare disease that previously was difficult to diagnose. Autoantibody screening as well as mutational analysis of the disease gene autoimmune regulator (AIRE) are important diagnostic tools for this life-threatening syndrome. OBJECTIVE: The objective of the study was to identify all patients with APS I in Norway and correlate their clinical features with their autoantibody profiles and mutations in the AIRE gene. PATIENTS: We identified 36 Norwegian patients from 24 families with APS I (20 males, 16 females) during a nationwide survey for patients with Addison's disease and polyendocrine syndromes, seven of them only after their death. RESEARCH DESIGN AND METHODS: Clinical data were collected from questionnaires and patient records. AIRE mutations were determined by DNA sequencing. Most autoantibodies were measured in RIAs against recombinant autoantigens, but anti type I interferon (IFN) antibodies were titrated in ELISA or antiviral interferon neutralization assays. RESULTS: The prevalence of APS I in Norway was estimated to be about 1:90,000. Several patients exhibited a milder phenotype with few APS I disease components and onset only in late adolescent or adulthood. The others showed about the same distribution of disease components as reported in Finnish patients. Eleven different mutations were identified in the AIRE gene, six of these were novel, i.e. c.22C>T (p.Arg8Cys), c.290T>C (p.Leu97Pro), c.402delC (p.Ser135GlnfsX12), c.879 + 1G>A (p.IVS7 + 1G>A), c.1249dupC (p.Leu417ProfsX7), and c.1336T>G (p.Cys446Gly). The 13-bp deletion in exon 8 (c.967-979del13) was the most prevalent mutation, present in 23 of 48 (48%) of the alleles. The presence of neutralizing autoantibodies against IFN-omega was the most specific marker of APS I, being found in all but one Norwegian patient. Some other common APS I-associated autoantibodies appeared de novo during long-term follow-up of younger patients. CONCLUSIONS: Norwegian patients with APS I clinically resemble those from Finland and other European countries, but some have milder phenotypes. In total, six new mutations were identified in the Norwegian APS I patients. Anti type I IFN autoantibodies are easily detectable; their APS I specificity and persistently high titers render them reliable markers of APS I, even in prodromal or atypical cases. Both the clinical features and the AIRE mutations are more diverse in the Norwegian population than previously thought. PMID- 17118991 TI - Efficacy and safety of leptin-replacement therapy and possible mechanisms of leptin actions in patients with generalized lipodystrophy. AB - BACKGROUND: Lack of leptin is implicated in insulin resistance and other metabolic abnormalities in generalized lipodystrophy; however, the efficacy, safety, and underlying mechanisms of leptin-replacement therapy in patients with generalized lipodystrophy remain unclear. METHODS: Seven Japanese patients with generalized lipodystrophy, two acquired and five congenital type, were treated with the physiological replacement dose of recombinant leptin during an initial 4 month hospitalization followed by outpatient follow-up for up to 36 months. RESULTS: The leptin-replacement therapy with the twice-daily injection dramatically improved fasting glucose (mean +/- SE, 172 +/- 20 to 120 +/- 12 mg/dl, P < 0.05) and triglyceride levels (mean +/- SE, 700 +/- 272 to 260 +/- 98 mg/dl, P < 0.05) within 1 wk. The leptin-replacement therapy reduced insulin resistance evaluated by euglycemic clamp method and augmented insulin secretion at glucose tolerance test with different responses between acquired and congenital types. Improvement of the fatty liver was also observed. The efficacy and safety of the once-daily injection were comparable to those of the twice daily injection. The leptin-replacement therapy ameliorated macro- and microalbuminuria and showed no deterioration of neuropathy and retinopathy of these patients. The leptin-replacement therapy is beneficial to diabetic complications and lipodystrophic ones. Two patients developed antileptin antibodies but not neutralizing antibodies. The therapy was well tolerated, and its effects were maintained for up to 36 months without any notable adverse effects such as hypoglycemia, high blood pressure, or reduction of bone mineral density. CONCLUSIONS: The present study demonstrates the efficacy and safety of the long-term leptin-replacement therapy and possible mechanisms of leptin actions in patients with generalized lipodystrophy. PMID- 17118992 TI - Diminished glucocorticoid negative feedback in polydipsic hyponatremic schizophrenic patients. AB - CONTEXT: The mechanism and significance of diminished glucocorticoid negative feedback in schizophrenia is unknown but is more commonly observed in schizophrenic patients with primary polydipsia. Polydipsic patients, especially those who are also hyponatremic, exhibit other neuroendocrine abnormalities that have been linked to hippocampal pathology. OBJECTIVE: The objective of the study was to determine the effect of cortisol on plasma ACTH under conditions thought to be most sensitive to hippocampal influences. DESIGN: The design was repeated measures. SETTING: The study was conducted at an inpatient clinical research center. PARTICIPANTS: Participants included eight polydipsic hyponatremic and eight polydipsic normonatremic as well as six schizophrenic patients without water imbalance. Eight healthy community volunteers matched for age and gender were also studied. INTERVENTION: Metyrapone (750 mg) was administered orally at 1430 and 1900 h. Beginning at 1930 h, hydrocortisone was infused over 150 min at 0.03 mg/kg.h. Blood samples and other measures were obtained at 20-min intervals from 1850 to 2320 h. MAIN OUTCOME MEASURES: Plasma ACTH and cortisol were measured. RESULTS: ACTH levels did not decline significantly during the cortisol infusion in the polydipsic hyponatremic group. For any given level of cortisol, ACTH levels were higher in the hyponatremic group. Although levels declined after cortisol in the other three groups, the decline was greatest in patients without water imbalance. CONCLUSIONS: The marked impairment in glucocorticoid negative feedback in polydipsic hyponatremic schizophrenic patients is consistent with hippocampal mineralocorticoid dysfunction. PMID- 17118993 TI - Glucose requirements to maintain euglycemia after moderate-intensity afternoon exercise in adolescents with type 1 diabetes are increased in a biphasic manner. AB - CONTEXT: Exercise increases the risk of hypoglycemia in type 1 diabetes. OBJECTIVE: This study aimed to investigate how the amount of glucose required to prevent an exercise-mediated fall in glucose level changes over time in adolescents with type 1 diabetes. SETTING: The study took place at a tertiary pediatric referral center. DESIGN, PARTICIPANTS, AND INTERVENTION: Nine adolescents with type 1 diabetes mellitus (five males, four females, aged 16 +/- 1.8 yr, diabetes duration 8.2 +/- 4.1 yr, hemoglobin A1c 7.8 +/- 0.8%, mean +/- SD) were subjected on two different occasions to a rest or 45 min of exercise at 95% of their lactate threshold. Insulin was administered iv at a rate based on their usual insulin dose, with similar plasma insulin levels for both studies (82.1 +/- 19.0, exercise vs. 82.7 +/- 16.4 pmol/liter, rest). Glucose was infused to maintain euglycemia for 18 h. MAIN OUTCOME MEASURES: Glucose infusion rates required to maintain euglcycemia and levels of counterregulatory hormones were compared between rest and exercise study nights. RESULTS: Glucose infusion rates to maintain stable glucose levels were elevated during and shortly after exercise, compared with the rest study, and again from 7-11 h after exercise. Counterregulatory hormone levels were similar between exercise and rest studies except for peaks in the immediate postexercise period (epinephrine, norepinephrine, GH, and cortisol peaks: 375.6 +/- 146.9 pmol/liter, 5.59 +/- 0.73 nmol/liter, 71.9 +/- 14.8 mIU/liter, and 558 +/- 69 nmol/liter, respectively). CONCLUSIONS: The biphasic increase in glucose requirements to maintain euglycemia after exercise suggests a unique pattern of early and delayed risk for nocturnal hypoglycemia after afternoon exercise. PMID- 17118994 TI - Thyroid-stimulating hormone receptor messenger ribonucleic acid measurement in blood as a marker for circulating thyroid cancer cells and its role in the preoperative diagnosis of thyroid cancer. AB - CONTEXT: Thyroid cancer cells express TSH receptor (TSHR) mRNA, and its measurement in the circulation may be useful in the diagnosis/management of differentiated thyroid cancer (DTC). OBJECTIVE: Our objective was to assess the diagnostic value of circulating TSHR mRNA for preoperative detection of DTC in patients with thyroid nodules. PATIENTS: We measured TSHR mRNA levels by RT-PCR in 258 subjects: 51 healthy subjects and 207 patients (thyroid nodules, n = 180; recurrent thyroid cancer, n = 27) with fine-needle aspirations (FNA) and/or thyroid/neck surgery. Eighty-nine patients also had d-1 postoperative levels assessed. OUTCOME MEASURES: TSHR mRNA levels were compared with FNA cytology for cancer detection preoperatively and serum thyroglobulin and/or whole-body 131I scans postoperatively. RESULTS: Based on cytology/pathology, 88 patients had DTC and 119 had benign thyroid disease. The TSHR mRNA levels in cancer patients were significantly higher than in benign disease (P < 0.0001). At a cutoff value of 1.02 ng/microg total RNA, the TSHR mRNA correctly classified 78.7% of patients preoperatively (sensitivity = 72.0%; specificity = 82.5%). Of 131 patients with FNA and surgery, 51 were FNA positive (all cancer), 17 were FNA negative (15 benign, two cancer), and 63 were indeterminate. TSHR mRNA correctly diagnosed DTC in 16 of 24 (67%) and benign disease in 29 of 39 (74%) patients with indeterminate FNA (combined sensitivity = 90%; specificity = 80%). Combining TSHR mRNA and ultrasound features for follicular lesions correctly classified all follicular cancers and could have spared surgery in 31% of these patients with benign disease. TSHR mRNA has a short life in circulation, and normalized levels on postoperative d 1 correlated with disease-free status, whereas elevated levels predicted residual/metastatic disease. CONCLUSIONS: TSHR mRNA measured with FNA enhances the preoperative detection of cancer in patients with thyroid nodules, reducing unnecessary surgeries, and immediate postoperative levels can predict residual/metastatic disease. PMID- 17118995 TI - Obesity and sex steroid changes across puberty: evidence for marked hyperandrogenemia in pre- and early pubertal obese girls. AB - CONTEXT: Peripubertal obesity is associated with abnormal sex steroid concentrations, but the timing of onset and degree of these abnormalities remain unclear. OBJECTIVE: The objective of the study was to assess the degree of hyperandrogenemia across puberty in obese girls and assess overnight sex steroid changes in Tanner stage 1-3 girls. DESIGN: This was a cross-sectional analysis. SETTING: The study was conducted at general clinical research centers. SUBJECTS: Thirty normal-weight (body mass index for age < 85%) and 74 obese (body mass index for age >or= 95%) peripubertal girls. INTERVENTION: Blood samples (circa 0500-0700 h) were taken while fasting. Samples from the preceding evening (circa 2300 h) were obtained in 23 Tanner 1-3 girls. MAIN OUTCOME MEASURES: Hormone concentrations stratified by Tanner stage were measured. RESULTS: Compared with normal-weight girls, mean free testosterone (T) was elevated 2- to 9-fold across puberty in obese girls, whereas fasting insulin was 3-fold elevated in obese Tanner 1-3 girls (P < 0.05). Mean LH was lower in obese Tanner 1 and 2 girls (P < 0.05) but not in more mature girls. In a subgroup of normal-weight Tanner 1-3 girls (n = 17), mean progesterone (P) and T increased overnight 2.3- and 2.4 fold, respectively (P 100 pg/ml) were 40% in the entire series. CONCLUSIONS: CT screening of thyroid nodules is a highly sensitive test for early diagnosis of MTC, but confirmatory stimulation testing is necessary in most cases to identify true positive increases. PMID- 17119001 TI - Effect of human body weight changes on circulating levels of peptide YY and peptide YY3-36. AB - BACKGROUND: Recent findings suggest that low plasma peptide YY (PYY) levels may contribute to diet-induced human obesity and justify PYY replacement therapy. Although the pharmacological value of PYY is controversial, further study of the secretion of the precursor PYY(1-36) and the pharmacologically active PYY(3-36) is indicated to determine the potential role in energy balance regulation. AIM: Our objective was to determine the effects of acute and chronic changes in human body weight on circulating levels of the putative satiety hormone peptide YY. DESIGN: Total plasma PYY levels (PYY(1-36) + PYY(3-36)) were measured in 66 lean, 18 anorectic, 63 obese, and 16 morbidly obese humans. In addition, total PYY was measured in 17 of the obese patients after weight loss and in the 18 anorectic patients after weight gain. Fasting PYY(3-36) levels were measured in 17 lean and 15 obese individuals. RESULTS: Fasting total plasma PYY levels were highest in patients with anorexia nervosa (80.9 +/- 12.9 pg/ml, P < 0.05) compared with lean (52.4 +/- 4.6 pg/ml), obese (43.9 +/- 3.8 pg/ml), or morbidly obese (45.6 +/- 11.2 pg/ml) subjects. In obese patients, weight loss of 5.4% was associated with a 30% decrease in fasting total PYY plasma levels. In anorectic patients, weight gain had no effect on fasting PYY. PYY(3-36) levels did not differ between lean (96.2 +/- 8.6 pg/ml) and obese (91.5 +/- 6.9 pg/ml) subjects. CONCLUSION: Our findings do not support a role for abnormal circulating PYY in human obesity. We conclude that circulating PYY levels in humans are significantly elevated in anorexia nervosa and, given the controversially discussed anorectic effect of PYY, could theoretically contribute to that syndrome. PMID- 17119002 TI - Serum adiponectin is a predictor of coronary heart disease: a population-based 10 year follow-up study in elderly men. AB - CONTEXT: Cross-sectional and nested case-control studies indicate a relationship between adiponectin, obesity, and coronary heart disease (CHD). OBJECTIVE: Our objective was to investigate whether adiponectin could predict CHD in a population-based cohort of elderly men. DESIGN AND SETTING: From 1991-1995 a baseline investigation was carried out in 832 healthy men aged 70 yr in the Uppsala Longitudinal Study of Adult Men (ULSAM study). They were followed up to 10.4 yr using Swedish national registry data. The baseline investigation included anthropometry, blood pressure, smoking, serum lipids, a euglycemic insulin clamp, and fasting serum adiponectin. MAIN OUTCOME MEASURES: Main outcome measures were defined as death or first-time hospitalization for CHD (n = 116), recorded in the Cause of Death Registry or in the Hospital-Discharge Registry of the National Board of Health and Welfare, Sweden. Associations were analyzed using Cox's proportional hazards regression, presented as hazard ratios (HR) with 95% confidence intervals (CI) for 1 sd increase in the predictor variable. RESULTS: In a multivariable analysis including total cholesterol (HR, 1.24; CI, 1.02 1.50), high-density lipoprotein cholesterol (HR, 0.72; CI, 0.58-0.89), smoking (HR, 1.39; CI, 0.91-2.14), and systolic blood pressure (HR, 1.26; CI, 1.05-1.52), serum adiponectin was associated with lower risk for CHD (HR, 0.81; CI, 0.66 0.99). The association was independent of BMI and remained significant after adjustment for insulin sensitivity index. CONCLUSIONS: In this population-based cohort of healthy men, elevated serum levels of adiponectin were associated with a lower risk for CHD. Importantly, the association between adiponectin and CHD was independent of other well-known risk factors. PMID- 17119003 TI - Regulation of appetite in lean and obese adolescents after exercise: role of acylated and desacyl ghrelin. AB - CONTEXT: Increased physical activity is an integral part of weight loss programs in adolescents. We hypothesized that exercise could affect appetite-regulating hormones and the subjective desire to eat, which could partly explain the poor success rate of the existing interventions. OBJECTIVE: The objective of this study was to investigate prospectively the effects of exercise on acylated ghrelin (AG) and desacyl ghrelin (DG) concentrations and on appetite. SETTING: The setting for this study was a tertiary care center. PARTICIPANTS: Normal weight [NW; body mass index (mean +/- se), 20.7 +/- 0.5 kg/m2] and overweight (OW; body mass index, 32.4 +/- 1.7) male adolescents (n = 17/group, age 15.3 +/- 0.2 yr) were studied. INTERVENTION: Those studied participated in 5 consecutive days of aerobic exercise (1 h/d). MAIN OUTCOME: Changes in AG and DG concentrations and in appetite during a test meal were studied. RESULTS: Exercise did not significantly affect insulin sensitivity or body weight. Fasting total (AG and DG) ghrelin concentrations were lower in OW (600 +/- 33 pg/ml) compared with NW (764 +/- 33 pg/ml, P < 0.05) boys and were not affected by exercise. In contrast, there was a differential effect of exercise on both AG and DG (P or=100 cases which were also adjusted for sex and age were included. Fifteen case control studies and three prospective studies contributed to present analysis, nine of which had not been included in previous meta-analyses. We calculated the pooled relative risks (RR) with corresponding 95% confidence intervals (95% CI), as a weighted average of the estimated RRs by random-effect models. The pooled RR of all NHL among HCV-positive individuals was 2.5 (95% CI, 2.1-3.0), but substantial heterogeneity was found between studies and by study design. Pooled RRs were 2.5 (95% CI, 2.1-3.1) in case-control studies and 2.0 (95% CI, 1.8-2.2) in cohort ones. The strongest source of heterogeneity seemed to be the prevalence of HCV among NHL-free study subjects (RR for NHL among HCV-positive individuals 3.0 and 1.9, respectively, for >or=5% and <5% HCV prevalence). RRs were consistently increased for all major B-NHL subtypes, T-NHL, and primary sites of NHL presentation. Thus, previous suggestions that the RRs for HCV differed by NHL subtype were not confirmed in our meta-analysis. Associations weaker than with NHL were found between HCV infection and Hodgkin's lymphoma (RR, 1.5; 95% CI, 1.0 2.1) and multiple myeloma (RR, 1.6; 95% CI, 0.7-3.6), but they were based on much fewer studies than NHL. The etiologic fraction of NHL attributable to HCV varies greatly by country, and may be upward of 10% in areas where HCV prevalence is high. PMID- 17119032 TI - Body size, mammographic density, and breast cancer risk. AB - BACKGROUND: Greater weight and body mass index (BMI) are negatively correlated with mammographic density, a strong risk factor for breast cancer, and are associated with an increased risk of breast cancer in postmenopausal women, but with a reduced risk in premenopausal women. We have examined the associations of body size and mammographic density on breast cancer risk. METHOD: We examined the associations of body size and the percentage of mammographic density at baseline with subsequent risk of breast cancer among 1,114 matched case-control pairs identified from three screening programs. The effect of each factor on risk of breast cancer was examined before and after adjustment for the other, using logistic regression. RESULTS: In all subjects, before adjustment for mammographic density, breast cancer risk in the highest quintile of BMI, compared with the lowest, was 1.04 [95% confidence interval (CI), 0.8-1.4]. BMI was associated positively with breast cancer risk in postmenopausal women, and negatively in premenopausal women. After adjustment for density, the risk associated with BMI in all subjects increased to 1.60 (95% CI, 1.2-2.2), and was positive in both menopausal groups. Adjustment for BMI increased breast cancer risk in women with 75% or greater density, compared with 0%, increased from 4.25 (95% CI, 1.6-11.1) to 5.86 (95% CI, 2.2-15.6). CONCLUSION: BMI and mammographic density are independent risk factors for breast cancer, and likely to operate through different pathways. The strong negative correlated between them will lead to underestimation of the effects on risk of either pathway if confounding is not controlled. PMID- 17119033 TI - Correlates of repeat and recent mammography for women ages 45 to 75 in the 2002 to 2003 Health Information National Trends Survey (HINTS 2003). AB - BACKGROUND: Most national-level mammography data are for ever-had and most-recent screening. There are few national-level data on rates of repeat, on-schedule mammography, and on the prevalence and correlates of repeat mammography. It is also important to investigate the similarity of correlates for repeat and recent mammography. METHODS: Analyses were from data for women ages 45 to 75 in the 2002 to 2003 Health Information and National Trends Survey (HINTS 2003; N = 1,581). The two dependent variables were self-report of repeat mammography (two exams on schedule, based on an every-other-year interval) and recent mammography only (one mammogram within the past 2 years). RESULTS: The prevalence of recent mammography was 81.6% (95% confidence interval, 79.1-84.1) and for repeat mammography was 72.2% (95% confidence interval, 69.0-75.4). An access to care variable combining insurance coverage and regular source of care was the strongest sociodemographic correlate of both mammography indicators. Most other sociodemographic variables were not associated with mammography status. Five psychosocial/behavioral variables were associated with both mammography indicators (smoking status, attention to health information, knowledge of screening interval, worry about breast cancer, and recent mood status). Correlates were very similar for repeat and recent mammography. CONCLUSIONS: Although access to care had the strongest association with mammography, psychosocial and behavioral variables did better as a group than the sociodemographic variables. A standard set of such variables should be considered for all national surveys. PMID- 17119034 TI - Antibiotics and risk of breast cancer: up to 9 years of follow-up of 2.1 million women. AB - Antibiotic use has been associated with risk of breast cancer in previous reports. Using Cox proportional hazards analysis, we evaluated this association in 2,130,829 adult female subscribers of a health care program according to their receipt of prescriptions of antibiotics from outpatient pharmacies. Hormone use was taken into account. Altogether, 18,521 women developed breast cancer in up to 9.4 years of follow-up. Use of any antibiotic was associated with slightly increased risk [hazard ratio (HR), 1.14; 95% confidence interval (95% CI), 1.10 1.18] but there was little, if any, evidence of dose response, with HR of 1.17 (95% CI, 0.97-1.42) for >1,000 days of use compared with no use. The only two weakly associated antibiotic groups (HR >1.10 for >100 days of use) were tetracyclines and macrolides with HRs (95% CI) of 1.23 (1.11-1.36) and 1.16 (0.98 1.36), respectively. An association of lincosamides with breast cancer in an earlier, smaller database was not confirmed, but follow-up was too short in the present data for adequate evaluation. Medical record review suggested that acne and/or rosacea could be the underlying factor, associated with long-term antibiotic therapy and found by others to be associated with risk of breast cancer. Although causality cannot be ruled out, the observed associations of antibiotics overall, tetracyclines, and macrolides with breast cancer were weak and could be explained by uncontrolled confounding by the diseases being treated or by other factors. PMID- 17119035 TI - Prognostic value of PAI1 in invasive breast cancer: evidence that tumor-specific factors are more important than genetic variation in regulating PAI1 expression. AB - Plasminogen activator inhibitor-1 (PAI1) can promote cancer progression, and its protein expression in tumors is an independent indicator of poor prognosis in many forms of cancer. Here, we show that high PAI1 mRNA levels also predict for shorter overall survival in two independent breast cancer data sets, highlighting the importance of its transcriptional regulation. The -675insG (4G/5G) single nucleotide polymorphism in the PAI1 gene promoter has been shown to influence PAI1 transcription, with the 4G allele eliciting higher reporter gene expression in vitro and higher levels of circulating PAI1 in vivo. Nevertheless, its genotypic distribution in 2,539 British women with invasive breast cancer was virtually identical to that seen in 1,832 matched controls (P = 0.72), and annual mortality rates for 4G4G, 4G5G, and 5G5G cases were 2.6%, 2.8%, and 3.1% per year, respectively (P = 0.10). Thus, there was no association with breast cancer incidence or outcome, and in a separate set of breast cancers, the 4G/5G single nucleotide polymorphism showed no association with PAI1 mRNA expression (P = 0.85). By contrast, connective tissue growth factor (CTGF), which can regulate PAI1 expression in culture, was associated with PAI1 expression in three independent cohorts (P << 0.0001). In addition, PAI1 gene copy number differences in the tumors were correlated with PAI1 mRNA expression (P = 0.0005) and seemed to affect expression independently of CTGF. Thus, local factors, such as CTGF and genomic amplification, seem to be more important than germ line genetic variation in influencing PAI1 expression and its untoward effects in breast cancer. PMID- 17119036 TI - Genetic polymorphisms of the CYP19A1 gene and breast cancer survival. AB - The CYP19A1 protein (aromatase) plays a critical role in estrogen biosynthesis and thus may be related to the progression of breast cancer. We examined the association between CYP19A1 genetic polymorphisms and breast cancer survival in a cohort of 1,136 patients who were recruited as part of a population-based case control study in Shanghai, China from 1996 to 1998 and who has donated a DNA sample to the study. Patients were followed for cancer recurrence and mortality through July 2005. Nineteen haplotype tagging single-nucleotide polymorphisms (SNP) in the CYP19A1 gene were evaluated. For each of the five SNPs located in haplotype block 2, patients homozygous for the minor alleles had a reduced 5-year disease-free survival rate compared with those carrying the major allele. The age adjusted hazard ratios (HR) and 95% confidence intervals (95% CI) were 1.5 (1.1 2.1), 2.1 (1.2-3.6), 1.5 (1.1-2.0), 1.4 (1.0-2.0), and 1.4 (1.0-2.0) for hCV1664178, rs12900137, rs730154, rs936306, and rs1902586, respectively. Haplotype analyses showed that the haplotype CCCTA (all minor alleles of the five SNPs in block 2) was associated with decreased disease-free survival (HR, 1.9; 95% CI, 1.1-3.3). The nonsynonymous SNP, rs700519 (Arg264Cys), located in haplotype block 4, was also associated with breast cancer survival. The age adjusted HR for the Cys/Cys (T/T) genotype was 2.2 (95% CI, 1.2-4.1) for overall survival and 2.1 (95% CI, 1.1-3.9) for disease-free survival, compared with those carrying the Arg (C) allele. These results suggest that polymorphisms in the CYP19A1 gene may have effects on breast cancer prognosis. PMID- 17119037 TI - Effects of parity on pregnancy hormonal profiles across ethnic groups with a diverse incidence of breast cancer. AB - Epidemiologic evidence suggests that a full-term pregnancy may affect maternal risk of breast cancer later in life. The objective of this cross-sectional study was to compare circulating levels of maternal hormones affecting breast differentiation (human chorionic gonadotropin and prolactin) and proliferation [alpha-fetoprotein, insulin-like growth factor I (IGF-I), and estradiol] between women at a low to moderate risk (Asians and Hispanics), as compared with women at a high risk for breast cancer (Caucasians and African-Americans). Between May 2002 and December 2004, a total of 586 pregnant women were approached during a routine prenatal visit. Among them, 450 women (206 Caucasian, 126 Asian, 88 Hispanic, and 30 African-American) met the inclusion criteria and signed the informed consent. Only singleton pregnancies were considered. Blood samples were drawn during the second trimester of pregnancy. Laboratory analyses were done using the IMMULITE 2000 immunoassay system. Gestational age standardized mean levels of estradiol, IGF-I, and prolactin were significantly higher in Hispanic women compared with Caucasian women. Mean concentration of IGF-I was significantly higher in African-American women compared with Caucasian and Asian women. No significant differences in pregnancy hormone levels were observed between Caucasian and Asian (predominantly second-generation Chinese) women in this study. Irrespective of ethnicity, women who had their first pregnancy had substantially higher mean levels of alpha-fetoprotein, human chorionic gonadotropin, estradiol, and prolactin compared with women who previously had at least one full-term pregnancy. These data suggest that circulating pregnancy hormone levels may explain some of the ethnic differences in breast cancer risk. PMID- 17119038 TI - CYP17 genotypes differ in salivary 17-beta estradiol levels: a study based on hormonal profiles from entire menstrual cycles. AB - Variation in the levels of sex-steroid hormones results from differences in developmental conditions, adult lifestyle, and genetic polymorphism. Genes involved in sex-steroid biosynthesis have been implicated to influence levels of hormones in premenopausal women, but the results were inconclusive. We tested variation among women in levels of salivary estradiol (E(2)) corresponding to CYP17 genotypes. CYP17 encodes cytochrome P450c17alpha, which mediates two enzymes important in E(2) synthesis. In contrast to the earlier studies that relied on one or a few samples for assessing the E(2) levels of an individual woman, our study is based on daily collected saliva samples for one entire menstrual cycle. Sixty Polish women, ages 24 to 36 years, with regular menstrual cycles and no reported fertility problems participated in the study. Women with A2/A2 genotype had 54% higher mean E(2) levels than women with A1/A1 genotype (P = 0.0001) and 37% higher than women with A1/A2 genotype (P = 0.0008). Heterozygous A1/A2 women had 13 % higher E(2) levels than homozygous A1/A1 women (but this difference was significant only in a nonparametric test). Levels of E(2) during the day with highest E(2) (day -1) were 72% higher in A2/A2 compared with A1/A1 (P = 0.01) and 52 % higher compared with A1/A2 (P = 0.03). Our results suggest that CYP17 genotype may serve as a biomarker of endocrine function in women of reproductive age. (Cancer Epidemiol Biomarkers Prev 2006;15(11):2131-5). PMID- 17119039 TI - Reproductive factors, oral contraceptive use, and human papillomavirus infection: pooled analysis of the IARC HPV prevalence surveys. AB - High parity, early age at first full-term pregnancy (FTP), and long-term oral contraceptive (OC) use increase cervical cancer risk, but it is unclear whether these variables are also associated with increased risk of acquisition and persistence of human papillomavirus (HPV) infection, the main cause of cervical cancer. Information on reproductive and menstrual characteristics and OC use were collected from 14 areas worldwide, among population-based, age-stratified random samples of women aged 15 years or older. HPV testing was done using PCR-based enzyme immunoassay. Unconditional logistic regression was used to estimate the odds ratios (OR) of being HPV-positive according to reproductive and menstrual factors and corresponding 95% confidence intervals (CI). When more than two groups were compared, floating CIs (FCI) were estimated. A total of 15,145 women (mean age, 40.9 years) were analyzed. Women with >or=5 FTPs (OR, 0.90; 95% FCI, 0.76-1.06) showed a similar risk of being HPV-positive compared with women with only one FTP (OR, 1.00; 95% FCI, 0.86-1.16). However, nulliparous women showed an OR of 1.40 (95% CI, 1.16-1.69) compared with parous women. Early age at first FTP was not significantly related to HPV positivity. HPV positivity was similar for women who reported >or=10 years of use of OCs (OR, 1.16; 95% FCI, 0.85-1.58) and never users of OCs (OR, 1.00; 95% FCI, 0.90-1.12). Our study suggests, therefore, that high parity, early age at first FTP, and long-term OC use are not associated with HPV prevalence, but rather these factors might be involved in the transition from HPV infection to neoplastic cervical lesions. PMID- 17119040 TI - Cervical cancer screening among women in metropolitan areas of the United States by individual-level and area-based measures of socioeconomic status, 2000 to 2002. AB - BACKGROUND: Few studies have examined cancer screening among women residing in metropolitan areas in relation to both individual-level and area-based measures of socioeconomic status (SES). To learn more, we examined self-reported rates of Papanicolaou (Pap) testing among women living in metropolitan areas in relation to individual-level measures of SES (household income and education), and area based measures of SES (percentage of residents living in poverty, percentage with low education, and percentage working class). METHODS: Data were obtained from women who were interviewed by telephone during 2000 and 2002 as part of the Behavioral Risk Factor Surveillance System (BRFSS). Self-reported county of residence was used to classify respondents as residents of metropolitan statistical areas. Only BRFSS respondents who resided in 35 metropolitan statistical areas with a population of >or=1.5 million in 2000 were included in this analysis. Analyses were limited to women ages >or=18 years with no history of hysterectomy (n = 49,231). Area-based measures of SES were obtained by using county-level information from the 2000 U.S. Census. RESULTS: Only 75.4% [95% confidence interval (95% CI), 73.8-77.1%] of 3,947 women ages >or=18 years who had a reported household income of <15,000 dollars per year had received a Pap test in the previous 3 years, compared with 92.2% (95% CI, 91.2-93.1%) of 18,698 women with a household income of >or=50,000 dollars. Overall, 77.5% (95% CI, 75.7 79.3%) of women without a high school education had received a Pap test compared with 91.7% (91.0-92.3%) of college graduates. In multivariate analysis, we also found education level to be positively associated with Pap testing rates, especially among women residing in areas where a relatively low percentage of residents had a low education level (P < 0.0001). CONCLUSIONS: Individual-level measures of SES may be modified by county-level measures of SES. Analyses of cancer screening rates by measures of income, educational attainment, and other factors may help health officials to better direct their finite resources to areas of greatest need. PMID- 17119041 TI - Gonorrhea, syphilis, clinical prostatitis, and the risk of prostate cancer. AB - BACKGROUND: Although previous case-control studies have observed positive associations among gonorrhea, syphilis, clinical prostatitis, and prostate cancer, many may have been susceptible to recall and interviewer biases due to their retrospective designs. Therefore, to investigate these associations without concerns of recall and interviewer biases, we conducted a large, prospective investigation in the Health Professionals Follow-up Study. METHODS: In 1992, participants were asked to report their histories of gonorrhea, syphilis, and clinical prostatitis by mailed questionnaire. Prostate cancer diagnoses were ascertained by self-report on the 1994 and each subsequent biennial follow-up questionnaire and confirmed by medical record review. RESULTS: Of the 36,033 participants in this analysis, 2,263 were diagnosed with prostate cancer between the date of return of the 1992 questionnaire and 2002. No association was observed between gonorrhea [adjusted relative risk (RR), 1.04; 95% confidence interval (95% CI), 0.79-1.36] or syphilis (RR, 1.06; 95% CI, 0.44-2.59) and prostate cancer. Overall null results were also observed between clinical prostatitis and prostate cancer (RR, 1.08; 95% CI, 0.96-1.20), although a significant positive association was observed among younger men (<59 years) screened for prostate cancer (RR, 1.49; 95% CI, 1.08-2.06; P(interaction) = 0.006). CONCLUSIONS: Gonorrhea and syphilis do not seem to be risk factors for prostate cancer in this cohort of men with a lower burden of sexually transmitted infections. Clinical prostatitis is also unlikely to be a risk factor, although possible roles for prostatitis in younger men and asymptomatic prostatic infection and inflammation cannot be ruled out. PMID- 17119042 TI - Family history, perceived risk, and prostate cancer screening among African American men. AB - BACKGROUND: Many African American men have two major risk factors for prostate cancer. By ethnicity alone, they have twice the risk of Euro-American men of developing prostate cancer. Having a family history (brother or father with prostate cancer) also doubles their risk. The major hypotheses tested in this study are that men with a family history perceive their risk to be higher, are more worried about getting prostate cancer, and are more likely to have used cancer screening tests than men without such a history. METHODS: A sample of 208 African American men, ages 40 to 74 years, were recruited through relatives or friends whose prostate cancer diagnosis was reported to the California Cancer Registry during the years 1997 to 2001 and from churches and African American social groups. Following a screening interview to determine eligibility, 88 men with self-reported, first-degree family history of prostate cancer and 120 without such history were interviewed by telephone. Logistic regression was used to create models of perceived risk, prostate cancer worries, receipt of a digital rectal exam, and/or prostate-specific antigen (PSA) testing. RESULTS: Men with a self-reported family history of prostate cancer did not perceive their risk as higher than men without a family history, nor did they report more cancer worries. They were more likely to report having a recent PSA test, but not a digital rectal exam. Having a higher than average perceived risk was associated with younger age, a college education, and lower mental well-being, and reporting more prostate cancer worries and being more likely to have had a recent PSA test. CONCLUSIONS: Although there continues to be controversy about PSA testing, these data suggest that African American men at above-average risk are inclined to be screened. PMID- 17119043 TI - A prospective investigation of height and prostate cancer risk. AB - Greater adult height, which reflects a combination of early nutrition, exposure to androgens, growth hormones, and other factors during growth and development, as well as heredity, has been associated with increased prostate cancer risk in several observational studies, but findings have been inconsistent. We examined this relationship in the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study cohort. At baseline, 29,119 Finnish male smokers 50 to 69 years old had height and weight measured by trained personnel, provided information on demographic, smoking, medical, and other characteristics, and completed an extensive diet history questionnaire. A total of 1,346 incident prostate cancer cases were identified during a follow-up period of up to 17.4 years (median, 14.1 years). In age-adjusted Cox proportional hazards models, the hazard ratios and 95% confidence intervals for prostate cancer according to increasing quintiles of height [178 cm] were 1.00 (reference), 1.11 (0.93-1.32), 1.11 (0.95-1.31), 1.30 (1.01-1.55), and 1.14 (0.96-1.35); P(trend) = 0.04. In analyses stratified by disease stage (available for 916 cases), a strong dose-response relationship was observed between greater height and advanced, but not earlier-stage, disease [tumor-node-metastasis stage III-IV, hazard ratio and 95% confidence interval for increasing quintiles of height: 1.77 (1.18-2.65), 1.82 (1.25-2.65), 1.93 (1.29-2.90), and 2.02 (1.37-2.97); P(trend) = 0.0008, P(interaction) = 0.002]. Our study provides additional evidence that increased height is a risk factor for prostate cancer and suggests that taller men are particularly susceptible to advanced disease. PMID- 17119044 TI - Educating African American men about the prostate cancer screening dilemma: a randomized intervention. AB - BACKGROUND: Until there is a definitive demonstration that early diagnosis and treatment of prostate cancer reduces disease-related mortality, it is imperative to promote informed screening decisions by providing balanced information about the potential benefits and risks of prostate cancer screening. Within a community/academic collaboration, we conducted a randomized trial of a printed booklet and a videotape that were designed for African American (AA) men. The purpose of the trial was to determine the effect of the interventions on knowledge, decisional conflict, satisfaction with the screening decision, and self-reported screening. METHODS: Participants were 238 AA men, ages 40 to 70 years, who were members of the Prince Hall Masons in Washington, DC. Men were randomly assigned to the (a) video-based information study arm, (b) print-based information study arm, or (c) wait list control study arm. Intervention materials were mailed to men at home. Assessments were conducted at baseline, 1 month, and 12 months postintervention. Multivariate analyses, including ANCOVA and logistic regression, were used to analyze group differences. RESULTS: The booklet and video resulted in a significant improvement in knowledge and a reduction in decisional conflict about prostate cancer screening, relative to the wait list control. Satisfaction with the screening decision was not affected by the interventions. Self-reported screening rates increased between the baseline and the 1-year assessment, although screening was not differentially associated with either of the interventions. In exploratory analyses, prostate-specific antigen testing at 1 year was more likely among previously screened men and was associated with having low baseline decisional conflict. CONCLUSIONS: This study represents one of the first randomized intervention trials specifically designed to address AA men's informed decision making about prostate cancer screening. We have developed and evaluated culturally sensitive, balanced, and disseminable materials that improved knowledge and reduced decisional conflict about prostate cancer screening among AA men. Due to the high incidence and mortality rates among AA men, there is a need for targeted educational materials, particularly materials that are balanced in terms of the benefits and risks of screening. PMID- 17119045 TI - Adolescent milk fat and galactose consumption and testicular germ cell cancer. AB - Recent case-control studies suggested that dairy product consumption is an important risk factor for testicular cancer. We examined the association between consumption of dairy products, especially milk, milk fat, and galactose, and testicular cancer in a population-based case-control study including 269 case and 797 controls (response proportions of 76% and 46%, respectively). Dietary history was assessed by food frequency questions for the index persons and through their mothers including diet 1 year before interview and diet at age 17 years. We used conditional logistic regression to calculate odds ratios as estimates of the relative risk (RR), 95% confidence intervals (95% CI), and to control for social status and height. The RR of testicular cancer was 1.37 (95% CI, 1.12-1.68) per additional 20 servings of milk per month (each 200 mL) in adolescence. This elevated overall risk was mainly due to an increased risk for seminoma (RR, 1.66; 95% CI, 1.30-2.12) per additional 20 milk servings per month. The RR for seminoma was 1.30 (95% CI, 1.15-1.48) for each additional 200 g milk fat per month and was 2.01 (95% CI, 1.41-2.86) for each additional 200 g galactose per month during adolescence. Our results suggest that milk fat and/or galactose may explain the association between milk and dairy product consumption and seminomatous testicular cancer. PMID- 17119046 TI - Glutathione S-transferase polymorphisms and the synergy of alcohol and tobacco in oral, pharyngeal, and laryngeal carcinoma. AB - Investigations of the ability of polymorphisms in the GSTM1, GSTT1, and GSTP1 genes to alter susceptibility to head and neck squamous cell carcinoma (HNSCC) have examined gene-environment interaction in their detoxification of tobacco associated carcinogens. Little work has been done to ask if these variant genes also modify the interaction of tobacco and alcohol in the development of HNSCC. To test this hypothesis, we conducted a case-control study, enrolling 692 incident cases of HNSCC and 753 population controls. Information about lifetime tobacco and alcohol use was ascertained through questionnaires, and genotypes for GSTM1, GSTT1, and GSTP1 were determined from constitutional DNA. Genotype frequencies were compared among cases and controls, and the association between genotypes and tobacco use was evaluated on cancer risk through logistic regression. Deletion of GSTM1 was associated with an increased risk for HNSCC [odds ratio (OR), 1.3; 95% confidence interval (95% CI), 1.0-1.6]. GSTT1 deletion was associated with a slight decreased HNSCC risk (OR, 0.8; 95% CI, 0.6-1.0). Among those with GSTM1 present, the OR of cancer for heavy smoking was 2.6 (95% CI, 1.6-4.3) compared with 4.2 for those with the GSTM1 deleted (95% CI, 2.6 6.7). The combination of consuming 10 to 20 alcohol drinks weekly and smoking >45 pack-years was associated with a 13-fold elevated risk (OR, 12.6; 95% CI, 4.0 40.2) among the GSTM1 deleted subjects compared with an OR of 3.6 (95% CI, 1.5 8.7) among the GSTM1 present individuals. These data (showing that the GSTM1 deletion affects on the tobacco and alcohol synergy) suggest that the interaction of these carcinogens is, at least in part, driven by alcohol, enhancing the carcinogenic action of tobacco smoke. PMID- 17119047 TI - A tailored smoking, alcohol, and depression intervention for head and neck cancer patients. AB - BACKGROUND: Smoking, alcohol use, and depression are interrelated and highly prevalent in patients with head and neck cancer, adversely affecting quality of life and survival. Smoking, alcohol, and depression share common treatments, such as cognitive behavioral therapy and antidepressants. Consequently, we developed and tested a tailored smoking, alcohol, and depression intervention for patients with head and neck cancer. METHODS: Patients with head and neck cancer with at least one of these disorders were recruited from the University of Michigan and three Veterans Affairs medical centers. Subjects were randomized to usual care or nurse-administered intervention consisting of cognitive behavioral therapy and medications. Data collected included smoking, alcohol use, and depressive symptoms at baseline and at 6 months. RESULTS: The mean age was 57 years. Most participants were male (84%) and White (90%). About half (52%) were married, 46% had a high school education or less, and 52% were recruited from Veterans Affairs sites. The sample was fairly evenly distributed across three major head and neck cancer sites and over half (61%) had stage III/IV cancers. Significant differences in 6-month smoking cessation rates were noted with 47% quitting in the intervention compared with 31% in usual care (P < 0.05). Alcohol and depression rates improved in both groups, with no significant differences in 6 month depression and alcohol outcomes. CONCLUSION: Treating comorbid smoking, problem drinking, and depression may increase smoking cessation rates above that of usual care and may be more practical than treating these disorders separately. PMID- 17119048 TI - Risk of squamous cell carcinoma of the upper aerodigestive tract in cancer-free alcoholic Japanese men: an endoscopic follow-up study. AB - Asian case-control studies have shown a strong relationship between the development of squamous cell carcinoma (SCC) of the esophagus and alcohol consumption combined with inactive aldehyde dehydrogenase-2 (ALDH2*1/*2), less active alcohol dehydrogenase-1B (ADH1B*1/*1), high mean corpuscular volume (MCV), and self-reported facial flushing in response to alcohol. However, little is known about whether these risk factors prospectively influence cancer development in cancer-free alcoholics. Between 1993 and 2005, 808 Japanese alcoholic men diagnosed as cancer-free by an initial endoscopic screening examination received follow-up examinations ranging from 1 to 148 months (median, 31 months) later, and SCC of the upper aerodigestive tract was diagnosed in 53 of them (esophagus in 33 and oropharyngolarynx in 30). Cox proportional hazards analysis showed that the age-adjusted relative hazard for SCC was 11.55 [95% confidence interval (95% CI), 5.73-23.3] in ALDH2*1/*2 heterozygotes compared with ALDH2*1/*1 homozygotes, 2.02 (95% CI, 1.02-4.02) in ADH1B*1/*1 homozygotes compared with ADH1B*1/*2 heterozygotes or *2/*2 homozygotes, 2.64 (95% CI, 1.49-4.67) in patients with flushing compared with those who had never experienced flushing, 2.91 (95% CI, 1.63-5.20) in those with an MCV >or= 106 compared with those with an MCV < 106, 2.52 (95% CI, 1.22-5.22) in those who smoked >or=30 cigarettes per day compared with those who smoked 0 to 19 cigarettes per day, 7.26 (95% CI, 3.99-13.23) in those with esophageal dysplasia compared with those without distinct iodine unstained lesions >or=5 mm, and 0.28 (95% CI, 0.09-0.85) in those with body mass index >or= 23.2 (highest quartile) compared with those with body mass index < 19.0 (lowest quartile). These predictors are useful for selecting appropriately patients for careful follow-up examinations. PMID- 17119049 TI - Genes involved in DNA repair and nitrosamine metabolism and those located on chromosome 14q32 are dysregulated in nasopharyngeal carcinoma. AB - Polymorphisms in nitrosamine metabolism, DNA repair, and immune response genes have been associated with nasopharyngeal carcinoma (NPC). Studies have suggested chromosomal regions involved in NPC. To shed light on NPC etiology, we evaluated host gene expression patterns in 31 NPC and 10 normal nasopharyngeal tissue specimens using the Affymetrix Human Genome U133 Plus 2.0 Array. We focused on genes in five a priori biological pathways and chromosomal locations. Rates of differential expression within these prespecified lists and overall were tested using a bootstrap method. Differential expression was observed for 7.6% of probe sets overall. Elevations in rate of differential expression were observed within the DNA repair (13.7%; P = 0.01) and nitrosamine metabolism (17.5%; P = 0.04) pathways. Differentially expressed probe sets within the DNA repair pathway were consistently overexpressed (93%), with strong effects observed for PRKDC, PCNA, and CHEK1. Differentially expressed probe sets within the nitrosamine metabolism pathway were consistently underexpressed (100%), with strong effects observed for NQ01, CYP2B6, and CYP2E1. No significant evidence of increases in rate of differential expression was seen within the immune/inflammatory pathway. A significant elevation in rate of differential expression was noted for chromosome 4p15.1-4q12 (13.0%; P = 0.04); both overexpression and underexpression were evident (38% and 62%, respectively). An elevation in the rate of differential expression on chromosome 14q32 was observed (11.3%; P = 0.06) with a consistent pattern of gene underexpression (100%; P < 0.0001). These effects were similar when excluding late-stage tumors. Our results suggest that nitrosamine activation and DNA repair are important in NPC. The consistent down-regulation of expression on chromosome 14q32 suggests loss of heterozygosity in this region. PMID- 17119050 TI - Association between nonsteroidal anti-inflammatory drug use and the incidence of lung cancer in the Iowa women's health study. AB - BACKGROUND: Previous studies have suggested that use of aspirin or other nonsteroidal anti-inflammatory drugs (NSAIDs) may be associated with reduced risk of lung cancer, but the data are inconsistent and are limited particularly with respect to the effects of aspirin, separate from other NSAIDs. METHODS: The Iowa Women's Health Study is a prospective cohort of 41,836 Iowa women ages 55 to 69 years old at baseline in 1986. NSAID use was assessed in 1992. Over 10 years of follow-up, 403 incident cases of lung cancer were identified. The association of incident lung cancer with current use of aspirin or non-aspirin NSAIDs was analyzed after adjustment for lung cancer risk factors. Hazard ratios (HR) were estimated using multivariate COX proportional hazards regression. RESULTS: There were 27,162 women in the analytic cohort. After controlling for age, education, alcohol intake, pack-years, smoking status, body mass index, and total fruit intake, the RR of women taking six or more aspirin weekly was 1.21 (95% confidence interval, 0.92-1.59). The HR was 1.23 for women taking six or more non aspirin NSAIDs weekly (95% confidence interval, 0.92-1.65). There was no statistically significant trend by frequency of use for either aspirin (P(trend) = 0.22) or non-aspirin NSAIDs (P(trend) = 0.53). Analyses by histologic type and smoking status yielded similar null results. Information on dosage and duration of use were not available for this analysis. CONCLUSION: These findings do not suggest that aspirin or other NSAIDs reduce risk of lung cancer in this cohort of postmenopausal women. PMID- 17119051 TI - Coexpression of fragile histidine triad and c-kit is relevant for prediction of survival in patients with small cell lung cancer. AB - BACKGROUND: In a retrospective analysis of 195 patients with small cell lung cancer (SCLC), we examined the prognostic value of a coexpression of fragile histidine triad (FHIT) protein and c-kit on patient's survival. METHODS: As assessed by immunohistochemistry using formalin-fixed, paraffin-embedded tissue sections, tumors of 195 patients with SCLC were evaluated for FHIT and c-kit coexpression. RESULTS: Coexpression of FHIT and c-kit was observed in 53.3%; a positive expression of either FHIT or c-kit was found in 40.5%. Complete lack of FHIT and c-kit (6.2%) was associated with a significantly shorter survival time for the patients with a mean of 122 +/- 45 days compared with 468 +/- 89 days for patients with lung cancer coexpressing FHIT and c-kit (P = 0.0011). The proportion of FHIT- and c-kit-positive cells within a tumor was also related to survival time. Patients with tumors with a proportion between 0% to 25% of FHIT- and c-kit-positive cells had the worst survival of 157 +/- 34 days compared with 496 +/- 95 days for patients showing >25% FHIT- and c-kit-positive cells (P = 0.0002). Further, variables associated with shorter survival times were low performance status, elevated lactate dehydrogenase level, and advanced tumor stage according to tumor-node-metastasis classification. Multivariate analysis using Cox regression model, including 11 variables, confirmed the prognostic significance of a combined expression of FHIT and c-kit next to tumor stage, performance status, and lactate dehydrogenase level. CONCLUSIONS: Differential FHIT and c-kit expression was of prognostic relevance for survival in patients with SCLC and therefore provide useful variables for therapeutic decisions. PMID- 17119052 TI - Polymorphisms of vitamin D receptor and survival in early-stage non-small cell lung cancer patients. AB - Our previous analysis suggested that surgery season in the summer time and high vitamin D intake are associated with improved survival in early-stage non-small cell lung cancer (NSCLC) patients. Here, we investigated the associations of vitamin D receptor (VDR) polymorphisms of Cdx-2 G>A, FokI C>T, and BsmI C>T with overall survival (OS) and recurrence-free survival (RFS) in 373 early-stage NSCLC patients. The data were analyzed using log-rank test and Cox proportional hazards models. The median follow-up time was 71 months (range, 0.1-140 months), with 186 deaths and 127 recurrences. There was no association between VDR polymorphisms and survival, overall or among adenocarcinoma patients. Among squamous cell carcinoma (SCC) patients, the G/A+A/A genotype group of the Cdx-2 polymorphism was associated with better OS: the 5-year OS rates were 41% [95% confidence interval (95% CI), 28-53] for the G/G and 55% (95% CI, 39-71) for the G/A+A/A genotypes, respectively (P = 0.04, log-rank test), with the adjusted hazard ratio of 0.56 (95% CI, 0.33-0.95) for G/A+A/A versus G/G. For the joint effects of the three polymorphisms, subjects with two or more "protective" alleles have better OS among SCC patients, with the adjusted hazard ratios of 0.20 (95% CI, 0.09 0.48), 0.40 (95% CI, 0.19-0.87), and 0.43 (95% CI, 0.19-0.97), respectively, for subjects with two, three, and four or more "protective" alleles when compared with subjects with zero or one "protective" allele (P(trend) = 0.71). Similar associations were found in haplotype analysis and for RFS among SCC patients. In conclusion, VDR polymorphisms may be associated with improved survival among SCC patients of early-stage NSCLC. PMID- 17119053 TI - Modeling human metabolism of benzene following occupational and environmental exposures. AB - We used natural spline (NS) models to investigate nonlinear relationships between levels of benzene metabolites (E,E-muconic acid, S-phenylmercapturic acid, phenol, hydroquinone, and catechol) and benzene exposure among 386 exposed and control workers in Tianjin, China. After adjusting for background levels (estimated from the 60 control subjects with the lowest benzene exposures), expected mean trends of all metabolite levels increased with benzene air concentrations from 0.03 to 88.9 ppm. Molar fractions for phenol, hydroquinone, and E,E-muconic acid changed continuously with increasing air concentrations, suggesting that competing CYP-mediated metabolic pathways favored E,E-muconic acid and hydroquinone below 20 ppm and favored phenol above 20 ppm. Mean trends of dose-specific levels (micromol/L/ppm benzene) of E,E-muconic acid, phenol, hydroquinone, and catechol all decreased with increasing benzene exposure, with an overall 9-fold reduction of total metabolites. Surprisingly, about 90% of the reductions in dose-specific levels occurred below about 3 ppm for each major metabolite. Using generalized linear models with NS-smoothing functions (GLM + NS models), we detected significant effects upon metabolite levels of gender, age, and smoking status. Metabolite levels were about 20% higher in females and decreased between 1% and 2% per year of life. In addition, levels of hydroquinone and catechol were greater in smoking subjects. Overall, our results indicate that benzene metabolism is highly nonlinear with increasing benzene exposure above 0.03 ppm, and that current human toxicokinetic models do not accurately predict benzene metabolism below 3 ppm. Our results also suggest that GLM + NS models are ideal for evaluating nonlinear relationships between environmental exposures and levels of human biomarkers. PMID- 17119054 TI - Gene expression patterns distinguish colonoscopically isolated human aberrant crypt foci from normal colonic mucosa. AB - Aberrant crypt foci (ACF) are considered the earliest identifiable preneoplastic colonic lesions; thus, a greater understanding of the nature of genetic changes underlying the transformation of normal colonic mucosa (NM) into ACF may provide insight into the mechanisms of carcinogenesis. ACF were identified by indigo carmine spraying onto colonic mucosa during colonoscopy and isolated as standard pinch biopsies of the mucosal areas containing the ACF. RNAs isolated from ACF and matched NM biopsies from the ascending and descending colons of 13 patients were analyzed on arrays containing 9128 cDNAs. Thirty-four differentially expressed (P < 0.001) genes were found in a paired comparison of the ACF and NM samples, and 25 of 26 matched pairs of ACF and NM could be correctly classified in leave-one-out cross-validation. Differential expression for seven of eight genes was confirmed by real-time reverse transcription-PCR. Furthermore, ACF and NM samples, including six pairs of ACF and NM samples that had not previously been analyzed by array hybridization, can be correctly classified on the basis of the overexpression in ACF of three selected genes (REG4, SRPN-B5, and TRIM29) evaluated by real-time reverse transcription-PCR. In a separate analysis of 13 biopsy pairs from either ascending or descending colon, ACF and NM samples could also be correctly classified by the gene expression patterns. Analysis of gene expression differences in ACF from the ascending and descending colon versus NM samples indicates that ACF from these distinct colonic locations are converging toward similar gene expression profiles and losing differences in gene expression characteristic of NM from the ascending versus descending colon. PMID- 17119055 TI - Genetic variation in the nucleotide excision repair pathway and colorectal cancer risk. AB - Nucleotide excision repair (NER) enzymes are critical for the removal of bulky DNA adducts caused by environmental carcinogens, such as heterocyclic amines and polycyclic aromatic hydrocarbons, which are found in two putative risk factors for colorectal cancer, tobacco smoke and meat cooked at high temperature. To examine the association between common genetic variants in NER genes and the risk of colorectal cancer, we conducted a case-cohort study within the CLUE II cohort. Twenty-two single nucleotide polymorphisms in 11 NER genes were genotyped in 250 colorectal cancer cases and a subcohort of 2,224 participants. Incidence rate ratios (RR) and 95% confidence intervals (95% CI) were estimated using a modified Cox regression model and robust variance estimate. The ERCC6 1213G variant, which is thought to reduce NER capacity, was associated with an increased risk of colorectal cancer compared with the homozygous wild type (RR, 1.36; 95% CI, 1.00 1.86 and RR, 2.64; 95% CI, 1.53-4.58 for the RG and GG genotypes respectively with P(trend) = 0.0006). Having at least one XPC 492H allele was also associated with an increased risk of colorectal cancer (RR, 1.75; 95% CI, 1.20-2.57). When the combined effects of ERCC6 R1213G and XPC R492H were examined, the risk of colorectal cancer significantly increased with increasing number of variant alleles (P(trend) = 0.00003). Our study suggests that genetic polymorphisms in the NER genes, ERCC6 and XPC, may be associated with an increased risk of colorectal cancer. PMID- 17119056 TI - Somatic BRAF-V600E mutations in familial colorectal cancer. AB - The BRAF gene is mutated in 4% to 12% of unselected colorectal cancers, particularly those with high microsatellite instability and in premalignant lesions, such as serrated adenomas and hyperplastic polyps. However, it has been shown that activating BRAF mutations are almost never found in tumors from hereditary nonpolyposis colorectal cancer patients. To evaluate the role of oncogenic BRAF mutations in non-hereditary nonpolyposis colorectal cancer/non familial adenomatous polyposis familial colorectal cancer, we did a mutation screening of the most common BRAF mutation, the V600E mutation, in 194 colorectal tumors from patients with a positive family history of the disease. The BRAF V600E mutation was identified in 100% (8 of 8) of microsatellite-unstable tumors and in 9.7% (18 of 186) of microsatellite-stable tumors. Interestingly, families with extracolonic tumors showed a much higher mutation frequency (17.5%) compared with families with colonic cancer only (3.5%; P = 0.009). In addition, we studied colonoscopic results from 448 family members who had been under colonoscopic surveillance for several years. Subjects from families where the V600E mutation was identified had less adenomas compared with those from families where no BRAF mutation had been found (odds ratio, 8.5; 95% confidence interval, 1.1-64.6). These findings indicate that adenomas might be less important in the cancer development in the group of families with BRAF-V600E mutations and indirectly support a previous hypothesis that tumors might develop through the hyperplastic polyp-serrated adenoma pathway. In conclusion, our results suggest that BRAF V600E mutations are mainly involved in colorectal cancer families characterized by an increased risk of other common malignancies. PMID- 17119057 TI - New marker of colon cancer risk associated with heme intake: 1,4-dihydroxynonane mercapturic acid. AB - BACKGROUND: Red meat consumption is associated with an increased risk of colon cancer. Animal studies show that heme, found in red meat, promotes preneoplastic lesions in the colon, probably due to the oxidative properties of this compound. End products of lipid peroxidation, such as 4-hydroxynonenal metabolites or 8-iso prostaglandin-F(2)alpha (8-iso-PGF(2)alpha), could reflect this oxidative process and could be used as biomarkers of colon cancer risk associated with heme intake. METHODS: We measured urinary excretion of 8-iso-PGF(2)alpha and 1,4 dihydroxynonane mercapturic acid (DHN-MA), the major urinary metabolite of 4 hydroxynonenal, in three studies. In a short-term and a carcinogenesis long-term animal study, we fed rats four different diets (control, chicken, beef, and blood sausage as a high heme diet). In a randomized crossover human study, four different diets were fed (a 60 g/d red meat baseline diet, 120 g/d red meat, baseline diet supplemented with heme iron, and baseline diet supplemented with non-heme iron). RESULTS: DHN-MA excretion increased dramatically in rats fed high heme diets, and the excretion paralleled the number of preneoplastic lesions in azoxymethane initiated rats (P < 0.0001). In the human study, the heme supplemented diet resulted in a 2-fold increase in DHN-MA (P < 0.001). Urinary 8 iso-PGF(2)alpha increased moderately in rats fed a high heme diet (P < 0.0001), but not in humans. CONCLUSION: Urinary DHN-MA is a useful noninvasive biomarker for determining the risk of preneoplastic lesions associated with heme iron consumption and should be further investigated as a potential biomarker of colon cancer risk. PMID- 17119058 TI - The human leukocyte antigen class I region is associated with EBV-positive Hodgkin's lymphoma: HLA-A and HLA complex group 9 are putative candidate genes. AB - Various studies have indicated that the human leukocyte antigen (HLA) region is associated with Hodgkin's lymphoma. We recently showed a specific association of the HLA class I region with EBV-positive Hodgkin's lymphoma cases. One haplotype of two consecutive microsatellite markers (D6S265 and D6S510) was overrepresented in the patient group, whereas another haplotype was underrepresented. Here, we did fine mapping of this region of approximately 400 kb as a next step to find the causative single-nucleotide polymorphism(s) (SNP). To select candidate SNPs for screening the total study population, several known SNPs were determined by sequencing two individuals homozygous for either of the above-mentioned associated haplotypes. Seven SNPs displayed different alleles in these two individuals and were therefore analyzed in the total study population, including 238 Hodgkin's lymphoma patients and 365 family-based controls. All seven SNPs showed significant association with the EBV-positive patient group. Two of these SNPs were analyzed in a Scottish Hodgkin's lymphoma population and revealed significant associations as well. The associated SNPs are located nearby two putative candidate genes: HLA-A and HLA complex group 9. HLA-A represents the most interesting target because of its consistent expression in EBV-positive Hodgkin's lymphoma cases and its ability to present EBV-derived peptides to cytotoxic T cells. PMID- 17119059 TI - Interleukin-6-related genotypes, body mass index, and risk of multiple myeloma and plasmacytoma. AB - Interleukin-6 (IL-6) promotes normal plasma cell development and proliferation of myeloma cells in culture. We evaluated IL-6 genotypes and body mass index (BMI) in a case-control study of multiple myeloma and plasmacytoma. DNA samples and questionnaires were obtained from incident cases of multiple myeloma (n = 134) and plasmacytoma (n = 16; plasma cell neoplasms) ascertained from the Los Angeles County population-based cancer registry and from siblings or cousins of cases (family controls, n = 112) and population controls (n = 126). Genotypes evaluated included IL-6 promoter gene single nucleotide polymorphisms (SNP) at positions 174, -572, and -597; one variable number of tandem repeats (-373 A(n)T(n)); and one SNP in the IL-6 receptor (IL-6ralpha) gene at position -358. The variant allele of the IL-6 promoter SNP -572 was associated with a roughly 2-fold increased risk of plasma cell neoplasms when cases were compared with family [odds ratio (OR), 1.8; 95% confidence interval (95% CI), 0.7-4.7] or population controls (OR, 2.4; 95% CI, 1.2-4.7). The -373 9A/9A genotype was associated with a decreased risk compared with the most common genotype (OR for cases versus family controls, 0.4; 95% CI, 0.1-1.7; OR for cases versus population controls, 0.3; 95% CI, 0.1-0.9). No other SNPs were associated with risk. Obesity (BMI >or= 30 kg/m(2)) increased risk nonsignificantly by 40% and 80% when cases were compared with family controls or population controls, respectively, relative to persons with a BMI of <25 kg/m(2). These results suggest that IL-6 promoter genotypes may be associated with increased risk of plasma cell neoplasms. PMID- 17119060 TI - Risk group, skin lesion history, and sun sensitivity reliability in squamous cell skin cancer progression. AB - In studies of skin cancer, participants are often classified into risk groups based on self-reported history of sun exposure or skin characteristics. We sought to determine the reliability of self-reported skin characteristics among participants of a study to evaluate markers for nonmelanoma skin cancer (NMSC). Multiple questionnaires and screening protocols were administered over a 3-month period to individuals from three risk groups: existing sun damage on forearms but no visible actinic keratoses (n = 91), visible actinic keratoses (n = 38), and history of resected squamous cell skin cancer in the last 12 months (n = 35). We assessed consistency of risk group assignment between telephone screen and study dermatologist assignment, self-reported sun sensitivity (telephone recruitment form versus participant completed profile), and self-reported history of NMSC skin lesions (telephone recruitment form versus health history). There was substantial agreement between probable risk group and final assignment (kappa = 0.76; 95% confidence interval, 0.65-0.85) and agreement did not differ by gender. Agreement for self-reported sun sensitivity was moderate (kappa weighted = 0.46; 95% confidence interval, 0.36-0.56) with higher agreement for women. For self reported NMSC lesion history between two interviews, 24 days apart, kappa estimates ranged from 0.66 to 0.78 and were higher for women than men. Overall, there was evidence for substantial reproducibility related to risk group assignment and self-reported history of NMSC, with self-reported sun sensitivity being less reliable. In all comparisons, women had higher kappa values than men. These results suggest that self-reported measures of skin cancer risk are reasonably reliable for use in screening subjects into studies. PMID- 17119061 TI - Ethnic disparity in the relationship between obesity and plasma insulin-like growth factors: the multiethnic cohort. AB - Previous studies on the relationship between obesity and circulating insulin-like growth factor (IGF) hormones show inconsistent findings and have not considered the possibility of racial/ethnic-specific differences that may exist. We therefore examined the relationship between obesity status [as measured by body mass index (BMI)] and plasma levels of the IGF proteins, IGF-I, IGF-binding protein 3 (IGFBP-3), and the molar ratio of IGF-I/IGFBP-3 in Whites, African Americans, Latinos, Japanese Americans, and Native Hawaiians from the ongoing Hawaii and Los Angeles Multiethnic Cohort Study. We measured plasma IGF-I and IGFBP-3 by ELISA in a random sample of 811 Multiethnic Cohort participants (53% male, age range = 47-82 at blood draw). In a multivariate regression of IGF-I levels, we found a statistically significant interaction between race/ethnicity and obesity status (P = 0.005). Plasma IGF-I levels declined with increasing BMI most dramatically in Latinos and Japanese. This decline was attenuated in Whites and absent in African-American and Native Hawaiian subjects. In Japanese, the quadratic term (BMI(2)) was statistically significant in a multivariate model (P = 0.002). In Latinos, the adjusted least-squares mean IGF-I levels in ng/mL for BMI < 25, 25 to 29.99, and >or=30 were 184.6, 147.7, and 132.7, respectively. No interaction between race/ethnicity and BMI explained the plasma IGFBP-3 levels in these data. These results may help to resolve the uncertainty in the relationship between circulating IGF levels and obesity and highlight the potential importance of racial/ethnic-specific effects among these factors in explaining ethnic disparities in obesity-related cancers. PMID- 17119062 TI - Mammographic breast density, dense area, and breast area differences by phase in the menstrual cycle. AB - BACKGROUND: Mammographic breast density may be greater in the luteal phase (days 15-30) than the follicular phase (days 1-14) of the menstrual cycle; this may have implications for when mammography screening should occur. OBJECTIVE: Examine whether percent breast density, breast area, or dense area differ by menstrual phase. METHODS: We identified 204 premenopausal women with regular periods who were <55 years (mean = 45.0 years) and had two screening mammograms within 9 to 18 months, with one screening between days 9 and 14, and one screening between days 22 and 35 of the menstrual cycle. We measured percent breast density, breast area, and dense area using the Cumulus software. We used linear regression to test for differences in breast density, breast area, and dense area from follicular to luteal phase, adjusting for change in weight and time between exams. RESULTS: The mean (SD) percent breast density was 35.8% (21.3) in the follicular phase and 36.7% (21.3) in the luteal phase. Multivariable analyses showed small but not statistically significant increases in percent density [1.1%; 95% confidence interval (95% CI), -0.2% to 2.3%] and breast area (16.7 cm(2); 95% CI, -2.8 to 36.2) and a statistically significant increase in dense area (13.1 cm(2); 95% CI, 0.1-26.1) in the luteal compared with the follicular phase. CONCLUSIONS: Breast density, breast area, and dense area have small, but probably not clinically meaningful, increases in the luteal phase of the menstrual cycle. However, there are other factors that may differ by menstrual cycle phase that we were unable to assess (e.g., breast compression), which may ultimately influence mammographic sensitivity by menstrual cycle phase. PMID- 17119063 TI - Genetic polymorphisms in xenobiotic clearance genes and their influence on disease expression in hereditary nonpolyposis colorectal cancer patients. AB - BACKGROUND: Hereditary nonpolyposis colorectal cancer (HNPCC) is associated with germ-line mutations in DNA mismatch repair genes. There is considerable variation in disease expression that cannot be explained by genotype/phenotype correlation, which is likely to be the result of polymorphic modifier genes. One candidate group of modifiers is the xenobiotic clearance enzyme genes that encode CYP1A1, GSTM1, GSTT1, GSTP1, and NAT2. Alterations in these xenobiotic clearance genes can potentially influence the host response to carcinogen exposure and thereby alter cancer risk. We have investigated eight polymorphisms in xenobiotic clearance genes to assess the effect on the risk of disease in mutation positive HNPCC patients. METHODS: DNA samples from 220 mutation-positive HNPCC participants (86 Australian and 134 Polish) were genotyped for single nucleotide polymorphisms (SNP) in CYP1A1, GSTM1, GSTT1, GSTP1, and NAT2. The association between the SNPs and disease characteristics, disease expression and age of diagnosis of colorectal cancer (CRC), was tested with Pearson's chi(2) and Kaplan Meier survival analysis. RESULTS: The HNPCC population displays a significant difference in the genotype frequency distribution between CRC patients and unaffected mismatch repair gene mutation carriers for the CYP1A1 SNP where the CRC patients harbor more of the mutant genotype. CONCLUSIONS: Evidence from this study is not conclusive, but our data suggest that the CYP1A1 influences disease expression in individuals with HNPCC. PMID- 17119064 TI - Effect of mammography on breast cancer risk in women with mutations in BRCA1 or BRCA2. AB - Women who carry mutations in either the BRCA1 or BRCA2 genes are at risk for early-onset breast cancer and are recommended to begin screening mammography at age 25 to 30 years. Results of in vitro and animal studies suggest that BRCA1/BRCA2 mutation carriers are hypersensitive to ionizing radiation and possibly to radiation-induced breast cancer. This study was undertaken to investigate the association of low-dose radiation exposure from mammograms with breast cancer status in BRCA mutation carriers. One hundred sixty-two female mutation carriers provided information at time of genetic testing about exposure to mammograms before enrollment. Using unconditional logistic regression, breast cancer status was not associated with number of mammograms received before diagnosis (affected women) or ascertainment [unaffected women; adjusted odds ratio (OR), 0.94; P = not significant]. A larger group of 213 women provided information about lifetime number of mammograms. There was no association between mammogram exposure and risk in the group as a whole (adjusted OR, 1.04; P = not significant), although there was a modest association in BRCA1 carriers (adjusted OR, 1.08; P = 0.03). These findings indicate that screening mammography is unlikely to be associated with a large increase in breast cancer risk in this population. PMID- 17119065 TI - The methionine synthase polymorphism c.2756A>G alters susceptibility to glioblastoma multiforme. AB - Genetic polymorphisms of methionine metabolism, in particular methionine synthase (MTR) c.2756A>G (D919G) and methylenetetrahydrofolate reductase (MTHFR) c.677C>T (A222V), have been associated with various human cancers. We investigated MTR c.2756A>G, MTHFR c.677C>T, and a third polymorphism, transcobalamin 2 c.776C>G (P259R), for a potential association with the formation of glioblastoma multiforme. The MTR c.2756G allele was significantly underrepresented among 328 glioblastoma multiforme patients of Caucasian origin when compared with 400 population controls [patients AA/AG/GG: 0.72/0.26/0.02 and controls AA/AG/GG: 0.57/0.38/0.05, degrees of freedom = 2; chi(2) = 17.86 (Pearson); P < 0.001]. No association between glioblastoma multiforme and the two other polymorphisms was observed. PMID- 17119066 TI - Higher methylation levels in gastric mucosae significantly correlate with higher risk of gastric cancers. AB - BACKGROUND: Helicobacter pylori infection potently induces methylation of CpG islands in gastric mucosae, which is considered to decrease to a certain level after active H. pylori infection discontinues. Noncancerous gastric mucosae of H. pylori-negative cases with a gastric cancer had higher methylation levels than those of H. pylori-negative healthy individuals. Here, using cases with multiple gastric cancers, we analyzed whether the higher methylation levels correlated with the higher risk of gastric cancers. METHODS: Twenty-six healthy volunteers (HV), 30 cases with a single well-differentiated gastric cancer (S cases), and 32 cases with multiple well-differentiated gastric cancers (M cases) were recruited. H. pylori infection status was analyzed by the culture method. Methylation levels were quantified by real-time methylation-specific PCR of seven CpG islands. RESULTS: In H. pylori-negative individuals, significant increasing trends were present in the order of HV, S cases, and M cases for FLNc and HAND1 methylation levels (P < 0.01, Spearman's rank-order test). Furthermore, the FLNc methylation level of M cases was significantly higher than that of S cases (P < 0.01, t test). Even adjusted by the extent of gastric atrophy, the FLNc methylation level retained a significant increasing trend (P = 0.03). In contrast, methylation levels in H. pylori-positive individuals were increased to various degrees in all the three groups. CONCLUSIONS: In H. pylori-negative individuals, methylation levels in gastric mucosae significantly increased in cases with a single gastric cancer and more in cases with multiple gastric cancers. Quantitative analysis of methylation levels is a promising risk marker for gastric cancers. PMID- 17119067 TI - Methylenetetrahydrofolate reductase haplotype tag single-nucleotide polymorphisms and risk of breast cancer. PMID- 17119068 TI - APC E1317Q is not associated with Colorectal Cancer in a population-based case control study in Northern Israel. AB - A variant in the adenomatous polyposis coli gene, APC E1317Q, has been inconsistently associated with risk of colorectal cancer. We used data collected as a part of the Molecular Epidemiology of Colorectal Cancer study, a population based study of colorectal cancer in northern Israel (1,834 matched cases and controls), to evaluate the relationship between this variant and risk of sporadic colorectal cancer. There was no association between E1317Q and colorectal cancer [odds ratio (OR), 1.15; 95% CI, 0.65-2.02]. When the control sample was restricted to polyp-free controls, the OR was 0.87 (95% CI, 0.36-2.14), indicating that this result is unlikely to be due to nondifferential misclassification due to undiagnosed polyps. A meta-analysis including these data and prior published reports found a nonsignificant summary OR nearly identical to the association reported here in the Molecular Epidemiology of Colorectal Cancer case-control study. Although there are previously published reports addressing this question, due to the low frequency of the variant, many of these studies lack the power to estimate the risk in a meaningful way. Given the substantial size of our study and the consistency of our findings with the results of our meta-analyses, we conclude that it is unlikely that APC E1317Q is associated with a clinically meaningful risk of colorectal cancer. PMID- 17119069 TI - Assessing the performance of different high-density tiling microarray strategies for mapping transcribed regions of the human genome. AB - Genomic tiling microarrays have become a popular tool for interrogating the transcriptional activity of large regions of the genome in an unbiased fashion. There are several key parameters associated with each tiling experiment (e.g., experimental protocols and genomic tiling density). Here, we assess the role of these parameters as they are manifest in different tiling-array platforms used for transcription mapping. First, we analyze how a number of published tiling array experiments agree with established gene annotation on human chromosome 22. We observe that the transcription detected from high-density arrays correlates substantially better with annotation than that from other array types. Next, we analyze the transcription-mapping performance of the two main high-density oligonucleotide array platforms in the ENCODE regions of the human genome. We hybridize identical biological samples and develop several ways of scoring the arrays and segmenting the genome into transcribed and nontranscribed regions, with the aim of making the platforms most comparable to each other. Finally, we develop a platform comparison approach based on agreement with known annotation. Overall, we find that the performance improves with more data points per locus, coupled with statistical scoring approaches that properly take advantage of this, where this larger number of data points arises from higher genomic tiling density and the use of replicate arrays and mismatches. While we do find significant differences in the performance of the two high-density platforms, we also find that they complement each other to some extent. Finally, our experiments reveal a significant amount of novel transcription outside of known genes, and an appreciable sample of this was validated by independent experiments. PMID- 17119070 TI - The changing face of coeliac disease. PMID- 17119071 TI - One hundred years of telemedicine: does this new technology have a place in paediatrics? PMID- 17119072 TI - Searching for the Holy Grail of acute otitis media. PMID- 17119073 TI - Unilateral fixed dilated pupil in a well child. PMID- 17119075 TI - Teaching medical students to examine children. PMID- 17119076 TI - Atomoxetine-induced electrocardiogram changes. PMID- 17119077 TI - Developing critical mass and growing our own academics. AB - Academic paediatrics is an exciting and rewarding career path but is not immune to the problems of recruitment and retention currently affecting most branches of medicine. The Modernising Medical Careers initiative, with its explicit academic training path, offers an unparalleled opportunity to develop novel schemes that promote recruitment and retention. Coordinated action is required to define, publicise and support the new academic training programmes and to attract the best trainees into them. PMID- 17119078 TI - Resuscitation of general paediatrics in the UK. AB - "The report of my death was an exaggeration", said Mark Twain. For a dying specialty, general paediatrics has certainly been looking very healthy recently. It is timely to examine why our specialty was thought to be at such risk, and to explore why, although in many cases shocked and confused, it is well on the way to recovery. This article explores what is needed to keep it healthy to ensure that the general paediatrician is at the centre of the delivery of paediatrics in the UK. PMID- 17119079 TI - Magnetic resonance imaging in suspected septic arthritis can avoid unnecessary surgery. PMID- 17119080 TI - Is lumbar puncture necessary for evaluation of early neonatal sepsis? PMID- 17119083 TI - Does the use of calamine or antihistamine provide symptomatic relief from pruritus in children with varicella zoster infection? PMID- 17119084 TI - Is supplementary iron useful when preterm infants are treated with erythropoietin? PMID- 17119085 TI - Use of buccal midazolam in children. PMID- 17119086 TI - Hypernatraemic dehydration in breast-fed neonates. PMID- 17119087 TI - NICE guidelines and the epilepsies: how should practice change? PMID- 17119088 TI - Prejudice against test weighing, like tongue-tie division, should be re-assessed. PMID- 17119089 TI - Tongue-tie and infant feeding. PMID- 17119090 TI - Changes in guidance for prescription of controlled drugs. PMID- 17119091 TI - Banning mobile phones in hospitals: is there enough evidence? PMID- 17119092 TI - Chest x ray and high-resolution computed tomography in cystic fibrosis. PMID- 17119093 TI - Breast feeding and the Baby Friendly Initiative: is the training of doctors being ignored? PMID- 17119094 TI - Diagnostic value of skull x rays in infants with plagiocephaly. PMID- 17119095 TI - A safe solution. PMID- 17119096 TI - The burden of acute nasopharyngitis among adolescents. PMID- 17119098 TI - Role of metal ions in the tetraloop-receptor complex as analyzed by NMR. AB - Metal ions are critical for the proper folding of RNA, and the GAAA tetraloop receptor is necessary for the optimal folding and function of many RNAs. We have used NMR to investigate the role of metal ions in the structure of the tetraloop receptor in solution. The NMR data indicate native tertiary structure is formed under a wide range of ionic conditions. The lack of conformational adaptation in response to very different ionic conditions argues against a structural role for divalent ions. Nuclear Overhauser effects to cobalt hexammine and paramagnetic relaxation enhancement induced by manganese ions were used to determine the NMR structures of the tetraloop receptor in association with metal ions, providing the first atomic-level view of these interactions in the solution state. Five manganese and two cobalt hexammine ions could be localized to the RNA surface. The locations of the associated metal ions are similar, but not identical to, those of previously determined crystal structures. The sites of association are in general agreement with nonlinear Poisson-Boltzmann calculations of the electrostatic surface, emphasizing the general importance of diffusely associated ions in RNA tertiary structure. PMID- 17119099 TI - Heterodimerization regulates RNase MRP/RNase P association, localization, and expression of Rpp20 and Rpp25. AB - Rpp20 and Rpp25 are subunits of the human RNase MRP and RNase P endoribonucleases belonging to the Alba superfamily of nucleic acid binding proteins. These proteins, which bind very strongly to each other, transiently associate with RNase MRP. Here, we show that the Rpp20-Rpp25 heterodimer is resistant to both high concentrations of salt and a nonionic detergent. The interaction of Rpp20 and Rpp25 with the P3 domain of the RNase MRP RNA appeared to be strongly enhanced by their heterodimerization. Coimmunoprecipitation experiments demonstrated that only a single copy of each of these proteins is associated with the RNase MRP and RNase P particles in HEp-2 cells. Both proteins accumulate in the nucleoli, which in case of Rpp20 is strongly dependent on its interaction with Rpp25. Finally, the results of overexpression and knock-down experiments indicate that their expression levels are codependent. Taken together, these data indicate that the Rpp20-Rpp25 heterodimerization regulates their RNA-binding activity, subcellular localization, and expression, which suggests that their interaction is also crucial for their role in RNase MRP/P function. PMID- 17119100 TI - Conformational analysis of the telomerase RNA pseudoknot hairpin by Raman spectroscopy. AB - We have measured the temperature-dependent Raman spectra of two 30-mer ribonucleotides that represent the wild-type (WT) and dyskeratosis congenita (DKC) mutant (MT) GC (107-108) --> AG structures of the pseudoknot hairpin region of human telomerase RNA. We have used these structures, previously characterized by UV-melting and NMR, as a model system for our Raman investigation. We observe that Raman hypochromism of vibrational bands, previously assigned to specific bases or conformational RNA markers, reflect temperature-dependent alterations in the pentaloop and stem structures of these two oligonucleotides. We also observe that the intense nu(s)(O-P-O) band at 812 cm(-1) indicates the presence of A-form backbone structure at relatively low temperatures in both the WT and MT RNA sequences. The mutation induces a decrease in the intensity of the uridine (rU) band at 1244 cm(-1) associated with C2'-endo/anti ribose conformation in the pentaloop. Two transition temperatures (T(m) ) were determined from the analysis of Raman difference intensity-temperature profiles of the 1256 cm(-1) band, which is associated with vibrations of cytidine (rC) residues, in particular, the C2' endo/anti ribose conformation (T(m) 1 = 23.6 +/- 1.6 degrees C for WT and 19.7 +/ 2.8 degrees C for MT; T(m) 2 = 68.9 +/- 1.8 degrees C for WT and 70.9 +/- 1.1 degrees C for MT). From these results we can conclude that the DKC mutant 30-mer exhibits a lower stability in the pentaloop region and a slightly higher stability in the stem region than the WT 30-mer. This demonstrates that Raman bands, previously assigned to specific bases or conformational RNA markers, can be used to probe local structural features of the telomerase pseudoknot hairpin sequence. PMID- 17119101 TI - Immunization 1967-2006: implementation keeping pace with invention? PMID- 17119102 TI - Treatment effects of antidepressants in patients with post-stroke depression: a meta-analysis. AB - BACKGROUND: Appropriate treatment of post-stroke depression (PSD) is critically important, considering the negative impact of PSD. Data regarding the treatment efficacy of antidepressants in patients with PSD are conflicting, and the time dependent effects of antidepressant treatment in this population are unknown. OBJECTIVE: To systematically assess treatment effects of antidepressants in patients with PSD, incorporating data from recent studies. METHODS: A meta analysis of randomized placebo-controlled trials (RCTs) of antidepressants in patients with PSD was conducted, using published studies from 1984 to 2006. Outcome measures of antidepressant treatment included response rate, depression rating scale scores, recovery of neurologic impairments, and improvements in activities of daily living (ADLs) after stroke. The effect size was presented as rate difference (RD) and weighted mean difference for dichotomous outcomes and continuous outcomes, respectively. Pooled effect sizes were calculated by both fixed-effects and random-effects models. RESULTS: A total of 1320 patients who met inclusion criteria were identified from 16 RCTs. The pooled response rates in the active and placebo groups were 65.18% (234/359) and 44.37% (138/311), respectively. The pooled RD was 0.23 (95% CI 0.03 to 0.43), indicating a significantly higher response rate in the active group compared with the placebo group. From baseline to endpoint, patients in the active group had significantly greater improvement in depressive symptoms compared with patients in the placebo group. Longer duration of treatment was positively correlated with the degree of improvement in depressive symptoms (Spearman's correlation, [rho] = -0.93, p = 0.001). No consistent evidence was found for positive antidepressant effects on the recovery of neurologic impairments and improvements in ADLs. CONCLUSIONS: The results of this meta-analysis suggest that use of antidepressants among patients with a diagnosis of PSD is associated with improvement in depressive symptoms. Longer durations of antidepressant treatment may be associated with greater reductions in depressive symptoms. PMID- 17119103 TI - Allopurinol as adjuvant therapy in poorly responsive or treatment refractory schizophrenia. AB - OBJECTIVE: To review the available literature evaluating the effectiveness of allopurinol for poorly responsive or treatment refractory schizophrenia. DATA SOURCES: Searches of MEDLINE (1966-October 2006), the Cochrane Library, and International Pharmaceutical Abstracts (1970-October 2006) were conducted using the terms allopurinol and schizophrenia. Limits were set to select studies conducted in humans. STUDY SELECTION AND DATA EXTRACTION: All articles identified from the data sources were evaluated. All case reports or clinical trials located were included in the review. DATA SYNTHESIS: Dopamine has been implicated for many years in the pathophysiology of schizophrenia, and the typical antipsychotics, via blockade of dopaminergic neurotransmission, have provided relief for patients with positive symptoms. However, because dopamine blockade does not relieve all symptoms of schizophrenia, it is now evident that many neurotransmitters may be involved in the pathogenesis of schizophrenia. Therefore, atypical antipsychotics, which target multiple neurotransmitters, have emerged as first-line therapies. An evolving body of evidence also supports a purinergic hypothesis for schizophrenia. Increased adenosinergic transmission is thought to reduce the affinity of dopamine agonists for dopamine receptors. Allopurinol, a xanthine oxidase inhibitor, may increase circulating pools of adenosine and may ultimately have antipsychotic and anxiolytic effects. Growing evidence for use of allopurinol as adjunctive therapy has been reported in both case reports and small clinical trials. CONCLUSIONS: Clinical trials show that adjuvant allopurinol may provide benefit to patients who are poorly responsive to current treatments for schizophrenia. Allopurinol is well tolerated by most patients. However, larger, randomized clinical trials need to be performed to determine the magnitude of this benefit, whether allopurinol should be routinely used as adjuvant therapy to antipsychotics, and which patient population is most likely to benefit from allopurinol use. For patients with limited options, allopurinol in doses of 300 mg once or twice daily may improve psychotic symptoms, especially refractory positive symptoms. PMID- 17119104 TI - Effect of oral corticosteroids on chronic warfarin therapy. AB - BACKGROUND: A potential drug interaction exists between oral corticosteroids and warfarin, but there is limited documentation. OBJECTIVE: To evaluate the potential drug interaction between oral corticosteroids and long-term warfarin therapy. METHODS: A retrospective review was conducted of 387 medical records for active patients within an anticoagulation clinic. Inclusion criteria were stable anticoagulation therapy, short-term oral corticosteroid therapy, international normalized ratio (INR) recorded within 30 days prior to corticosteroid initiation (pre-INR), and INR recorded during corticosteroid therapy or within 14 days of discontinuation (post-INR). Patients were excluded if they had been started on any antibiotic or other drug with a probable interaction with warfarin at the same time as corticosteroid initiation. Thirty-two patient encounters met the predetermined inclusion and exclusion criteria. The primary outcome assessed was the difference between pre- and post-INR values. Secondary endpoints included bleeding events, emergency department (ED) visits, hospitalizations, and warfarin dose modifications. RESULTS: The mean difference between pre- and post-INR values was 1.24 (95% CI 0.86 to 1.62). Ninety-seven percent of the 32 patient encounters resulted in a change in their post-INR value, and 62.5% of patients had supratherapeutic INR values at the post-corticosteroid assessment. The majority of patients assessed had an elevation of their INR following concomitant use of warfarin and corticosteroids. The INR change was observed at a mean +/- SD of 6.7 +/- 3.3 days following the first dose of corticosteroid. Overall, 16 patients (50%) required a modification of their anticoagulation therapy during or following corticosteroid therapy. Only one adverse event of minor epistaxis was reported, and no ED visits or hospitalizations occurred as a consequence of the drug combination. CONCLUSIONS: Use of oral corticosteroids in patients on long term warfarin therapy may result in a clinically significant interaction, which requires close INR monitoring and possible warfarin dose reduction. PMID- 17119105 TI - Emerging therapies in the treatment of Clostridium difficile-associated disease. AB - OBJECTIVE: To describe emergent therapies, such as rifaximin, nitazoxanide, intravenous immunoglobulin (IVIG), tinidazole, tolevamer, and the possible use of a vaccine, in Clostridium difficile-associated disease (CDAD), one of the most common causes of diarrhea in hospitalized adults in North America. DATA SOURCES: A literature search was performed using MEDLINE (1996-October 2006), PubMed (1996 October 2006), abstracts from Infectious Diseases Society of America (September 2006) and International Conference on Antimicrobial Agents and Chemotherapy (September 2006), Internet (October 2006), Genzyme product Web site (October 2006), and Romark Laboratories Web site (October 2006) using the terms Clostridium difficile, rifaximin, nitazoxanide, intravenous immunoglobulin, tolevamer, vaccine, and tinidazole. STUDY SELECTION AND DATA EXTRACTION: Data presented in this article were selected based on clinical relevance and power of the studies. In vivo and in vitro studies supporting the use of drugs available for treatment of refractory CDAD were reviewed. Some of the information on new and emerging modalities was also included, although there were limited published data available. DATA SYNTHESIS: Clinical trials evaluating the use of nitazoxanide and tolevamer for the treatment of CDAD have been published. Tinidazole use is based on structural similarities to metronidazole; however, clinical trials have not been conducted and the cost of this agent may be a limiting factor. The use of rifaximin and IVIG will require randomized clinical trials to establish their place in therapy. Limited information in the literature suggests that a vaccine may be effective for CDAD prevention. CONCLUSIONS: CDAD is a debilitating disease with increasing treatment failure rates and recurrences using standard therapies. Clinicians need to look at other options to expand the available treatment arsenal in addition to placing a greater emphasis on prevention. PMID- 17119106 TI - Neuroleptic malignant syndrome with delayed onset of fever following risperidone administration. AB - OBJECTIVE: To report and discuss a case of neuroleptic malignant syndrome (NMS) with delayed onset of fever in a patient taking risperidone. CASE SUMMARY: A 59 year-old white female presented with progressive weakness, confusion, and disorientation 10 days after restarting risperidone 2 mg/day therapy for bipolar disorder. She had taken risperidone for several years prior to this episode and had stopped it for approximately 3 weeks; risperidone was discontinued on admission. The patient's creatine kinase (CK) level was elevated (901 IU/L; reference range 39-162) on admission and increased to 1991 IU/L the following day. She was initially afebrile and had no muscular rigidity. Elevated temperature (38.1 degrees C) did not occur until hospital day 2. The patient was successfully treated with diazepam, bromocriptine, and dantrolene and suffered no long-term sequelae. DISCUSSION: Other clinicians have reported atypical presentations of NMS in patients taking newer neuroleptic agents. Although this patient met diagnostic criteria for NMS, the hallmark symptoms of fever and muscle rigidity were delayed in onset. Also, the patient never remained febrile for more than 24 hours and her maximum temperature was only 38.6 degrees C. An objective causality assessment suggests that this case of NMS was probably related to restarting risperidone. CONCLUSIONS: Because of the life-threatening nature of this syndrome, clinicians should consider NMS in afebrile patients presenting with diaphoresis, changes in level of consciousness, mutism, tremors, tachycardia, leukocytosis, and elevated CK levels. PMID- 17119107 TI - Efficacy with high-dose aripiprazole after olanzapine-related metabolic disturbances. AB - OBJECTIVE: To report a case in which a patient tolerated and responded to high dose aripiprazole and lost weight after having been on olanzapine for several years; a secondary objective is to discuss the use of aripiprazole dosages at higher than those recommended in the product's labeling. CASE SUMMARY: A 57-year old man with a 30 year history of schizophrenia had been taking olanzapine for 4 years, with the dosage titrated to 20 mg/day, to control the psychosis. After he had gained significant weight with olanzapine (the highest was 102.7 kg), his treatment was switched to aripiprazole. The patient required a high dose of aripiprazole (60 mg/day) to achieve full control of the psychiatric symptoms, and during aripiprazole therapy, he lost the weight he had gained while on olanzapine, weighing 85.9 kg within 7 months after the therapy switch. DISCUSSION: Dosages of atypical antipsychotics higher than those recommended by the Food and Drug Administration are often used in clinical practice for refractory patients, despite the lack of evidence. The literature available on this subject is limited to small, double-blind trials; open-label trials; and case reports. Although certain patients may benefit from higher doses of atypical antipsychotics, the lack of evidence limits their use. CONCLUSIONS: High-dose aripiprazole (60 mg/day) was well tolerated and controlled this patient's symptoms effectively. In addition, he lost weight that was gained while being treated with olanzapine. High-dose aripiprazole may be beneficial and safe in refractory patients; however, large, double-blind, randomized clinical trials are needed. PMID- 17119108 TI - von Willebrand factor and factor VIII are independently required to form stable occlusive thrombi in injured veins. AB - von Willebrand factor (VWF) protects factor VIII (FVIII) from proteolysis and mediates the initial contact of platelets with the injured vessel wall, thus playing an important role in hemostasis and thrombosis. VWF is crucial for the formation of occlusive thrombi at arterial shear rates. However, with only a few conflicting studies published, the role of VWF in venous thrombosis is still unclear. Using gene-targeted mice, we show that in ferric chloride-injured veins platelet adhesion to subendothelium is decreased and thrombus growth is impaired in VWF(-/-) mice when compared with wild type (WT). We also observed increased embolization in the VWF(-/-) mice, which was due to lower FVIII levels in these mice as recombinant factor VIII (r-FVIII) restored thrombus stability. Despite normalization of blood clotting time and thrombus stability after r-FVIII infusion, the VWF(-/-) venules did not occlude. Transgenic platelets lacking the VWF receptor GPIbalpha extracellular domain showed decreased adhesion to injured veins. But, after a delay, all the injured venules occluded in these transgenic mice. Thus, VWF likely uses other adhesion receptors besides GPIbalpha in thrombus growth under venous shear conditions. Our studies document crucial roles for VWF and FVIII in experimental thrombosis under venous flow conditions in vivo. PMID- 17119109 TI - Engulfment of apoptotic cells by microvascular endothelial cells induces proinflammatory responses. AB - Circulating endothelial cells (CECs) have been detected in a variety of vascular disorders, but their interactions with healthy endothelium remain unknown. The aim of this study was to evaluate the response of human endothelial cells (ECs) to apoptotic or necrotic ECs in an in vitro model and to delineate pathogenetic pathways. Here we show that incubation of the human microvascular endothelial cell line (HMEC-1) with apoptotic ECs resulted in increased expression of chemokines and enhanced binding of leukocytes to HMEC-1 cells, whereas exposure of HMEC-1 cells to necrotic ECs caused no changes in leukocyte-binding affinity. Both apoptotic and necrotic cells were bound and engulfed by HMEC-1 cells and primary human umbilical vein endothelial cells (HUVECs). We therefore suggest that exposures to apoptotic and necrotic ECs induce different patterns of chemokine synthesis and leukocyte adhesion in healthy ECs. These data indicate that CECs are not only markers of vascular damage but may induce proinflammatory signals in the endothelium. PMID- 17119110 TI - Naive recirculating B cells mature simultaneously in the spleen and bone marrow. AB - We have recently demonstrated that IgD(hi) B cells can occupy an extravascular perisinusoidal niche in the bone marrow in addition to the well-established follicular niche in conventional secondary lymphoid organs. The spleen has long been considered to be the site at which newly formed B lymphocytes mature into IgD(hi) naive recirculating B cells, but the existence of mutant mice that have selectively lost mature B cells in the bone marrow prompted an examination of B cell maturation at this latter site. Following a single pulse of BrdU in intact mice, sequential labeling of more mature B-cell populations in the bone marrow suggested ongoing maturation at this site. Further evidence for B-cell maturation in the bone marrow was obtained from analyses of transitional B cells in splenectomized lymphotoxin alpha-deficient mice that lack all secondary lymphoid organs. In these mice, antibody-secreting cells recognizing multivalent antigens were also observed in the bone marrow following an intravenous microbial challenge. These data suggest that newly formed B cells mature into IgD(hi) B cells simultaneously in the spleen and the bone marrow and establish in a stringent manner that humoral immune responses can be initiated in situ in the bone marrow. PMID- 17119111 TI - Prophylactic administration of imatinib after hematopoietic cell transplantation for high-risk Philadelphia chromosome-positive leukemia. AB - Relapse occurs frequently after allogeneic hematopoietic cell transplantation (HCT) for treatment of high-risk Philadelphia chromosome-positive (Ph+) leukemia. Administration of imatinib early after HCT might provide an effective approach for preventing recurrent Ph+ leukemia, but the feasibility of this approach has not been systematically tested. Twenty-two patients, 15 with Ph+ acute lymphoblastic leukemia and 7 with high-risk chronic myelogenous leukemia, were enrolled in a prospective study and given imatinib from the time of engraftment until 365 days after HCT. Before day 90, adults (n = 19) tolerated a median average daily imatinib dose of 400 mg/d (range, 200-500 mg/d), and children (n = 3) tolerated 265 mg/m2/d (range, 200-290 mg/m2/ d). The most common adverse events related to imatinib administration were grade 1-3 nausea, emesis, and serum transaminase elevations. We conclude that imatinib can be safely administered early after myeloablative allogeneic HCT at a dose intensity comparable to that used in primary therapy. PMID- 17119112 TI - RhoH is important for positive thymocyte selection and T-cell receptor signaling. AB - RhoH is a small GTPase expressed only in the hematopoietic system. With the use of mice with targeted disruption of the RhoH gene, we demonstrated that RhoH is crucial for thymocyte maturation during DN3 to DN4 transition and during positive selection. Furthermore, the differentiation and expansion of DN3 and DN4 thymocytes in vitro were severely impaired. These defects corresponded to defective TCR signaling. Although RhoH is not required for TCR-induced activation of ZAP70 and ZAP70-mediated activation of p38, it is crucial for the tyrosine phosphorylation of LAT, PLCgamma1, and Vav1 and for the activation of Erk and calcium influx. These data suggest that RhoH is important for pre-TCR and TCR signaling because it allows the efficient interaction of ZAP70 with the LAT signalosome, thus regulating thymocyte development. PMID- 17119114 TI - Late cardiotoxicity after treatment for Hodgkin lymphoma. AB - We assessed cardiovascular disease (CVD) incidence in 1474 survivors of Hodgkin lymphoma (HL) younger than 41 years at treatment (1965-1995). Multivariable Cox regression and competing risk analyses were used to quantify treatment effects on CVD risk. After a median follow-up of 18.7 years, risks of myocardial infarction (MI) and congestive heart failure (CHF) were strongly increased compared with the general population (standardized incidence ratios [SIRs] = 3.6 and 4.9, respectively), resulting in 35.7 excess cases of MI and 25.6 excess cases of CHF per 10 000 patients/year. SIRs of all CVDs combined remained increased for at least 25 years and were more strongly elevated in younger patients. Mediastinal radiotherapy significantly increased the risks of MI, angina pectoris, CHF, and valvular disorders (2- to 7-fold). Anthracyclines significantly added to the elevated risks of CHF and valvular disorders from mediastinal RT (hazard ratios [HRs] were 2.81 and 2.10, respectively). The 25-year cumulative incidence of CHF after mediastinal radiotherapy and anthracyclines in competing risk analyses was 7.9%. In conclusion, risks of several CVDs are 3- to 5-fold increased in survivors of HL compared with the general population, even after prolonged follow up, leading to increasing absolute excess risks over time. Anthracyclines further increase the elevated risks of CHF and valvular disorders from mediastinal radiotherapy. PMID- 17119113 TI - A phase 1/2 trial of arginine butyrate and ganciclovir in patients with Epstein Barr virus-associated lymphoid malignancies. AB - Malignancies associated with latent Epstein-Barr virus (EBV) are resistant to nucleoside-type antiviral agents because the viral enzyme target of these antiviral drugs, thymidine kinase (TK), is not expressed. Short-chain fatty acids, such as butyrate, induce EBV-TK expression in latently infected B cells. As butyrate has been shown to sensitize EBV(+) lymphoma cells in vitro to apoptosis induced by ganciclovir, arginine butyrate in combination with ganciclovir was administered in 15 patients with refractory EBV(+) lymphoid malignancies to evaluate the drug combination for toxicity, pharmacokinetics, and clinical responses. Ganciclovir was administered twice daily at standard doses, and arginine butyrate was administered by continuous infusion in an intrapatient dose escalation, from 500 mg/(kg/day) escalating to 2000 mg/(kg/day), as tolerated, for a 21-day cycle. The MTD for arginine butyrate in combination with ganciclovir was established as 1000 mg/(kg/day). Ten of 15 patients showed significant antitumor responses, with 4 CRs and 6 PRs within one treatment cycle. Complications from rapid tumor lysis occurred in 3 patients. Reversible somnolence or stupor occurred in 3 patients at arginine butyrate doses of greater than 1000 mg/(kg/day). The combination of arginine butyrate and ganciclovir was reasonably well-tolerated and appears to have significant biologic activity in vivo in EBV(+) lymphoid malignancies which are refractory to other regimens. PMID- 17119115 TI - Mechanisms of regulation of CXCR4/SDF-1 (CXCL12)-dependent migration and homing in multiple myeloma. AB - The mechanisms by which multiple myeloma (MM) cells migrate and home to the bone marrow are not well understood. In this study, we sought to determine the effect of the chemokine SDF-1 (CXCL12) and its receptor CXCR4 on the migration and homing of MM cells. We demonstrated that CXCR4 is differentially expressed at high levels in the peripheral blood and is down-regulated in the bone marrow in response to high levels of SDF-1. SDF-1 induced motility, internalization, and cytoskeletal rearrangement in MM cells evidenced by confocal microscopy. The specific CXCR4 inhibitor AMD3100 and the anti-CXCR4 antibody MAB171 inhibited the migration of MM cells in vitro. CXCR4 knockdown experiments demonstrated that SDF 1-dependent migration was regulated by the P13K and ERK/ MAPK pathways but not by p38 MAPK. In addition, we demonstrated that AMD3100 inhibited the homing of MM cells to the bone marrow niches using in vivo flow cytometry, in vivo confocal microscopy, and whole body bioluminescence imaging. This study, therefore, demonstrates that SDF-1/CXCR4 is a critical regulator of MM homing and that it provides the framework for inhibitors of this pathway to be used in future clinical trials to abrogate MM trafficking. PMID- 17119116 TI - Gene-nutrient interactions among determinants of folate and one-carbon metabolism on the risk of non-Hodgkin lymphoma: NCI-SEER case-control study. AB - We previously reported a lower risk of non-Hodgkin lymphoma (NHL) associated with high consumption of vitamin B6 and methionine, dietary determinants of one-carbon metabolism. Evidence has linked genetic variants involved in one-carbon metabolism to NHL. We investigated 30 polymorphisms in 18 genes for their main effect on NHL among 1141 incident cases and 949 population-based controls and examined gene-nutrient interactions in a subgroup of 386 cases and 319 controls who provided detailed food-frequency information. Odds ratios (ORs) and 95% confidence intervals (CIs) were adjusted for age, sex, and race. We observed a decreased risk of NHL over-all with BHMTEx8+453A>T and increased risk with CBS Ex13+41C>T, FPGS Ex15-263T>C, and SHMT1 Ex12+138C>T and Ex12+236C>T. Furthermore, significant gene-nutrient interactions limited the protective association comparing high versus low vitamin B6 to FPGS Ex15-263T>C CC (OR = 0.22; 95% CL = 0.10-0.52), MTHFS IVS2-1411T>G TT/TG (OR = 0.54; 95% CI = 0.36-0.81), and MTR Ex26-20A>G AA (OR = 0.55; 95% CI = 0.35-0.86) genotypes, and the protective association of methionine to FTHFD Ex10-40G>TGG (OR = 0.63; 95% CI = 0.44-0.91), MTHFR Ex8-62A>C CC (OR = 0.13; 95% CI = 0.04-0.39), and MTRR Ex5+136T>CTT (OR = 0.67; 95% CI = 0.47-0.97) genotypes. Warranting replication, our finding of gene nutrient interactions in one-carbon metabolism supports their etiologic involvement in lymphomagenesis. PMID- 17119117 TI - The growth factor fusion construct containing B-lymphocyte stimulator (BLyS) and the toxin rGel induces apoptosis specifically in BAFF-R-positive CLL cells. AB - The cytokine B lymphocyte stimulator (BLyS) mediates its effect through cell surface receptors BAFF-R, TACI, and BCMA. BLyS receptors are expressed only on B cells and not present in other normal cells including normal T lymphocytes. Chronic lymphocytic leukemia (CLL) is a B-cell disease and CLL lymphocytes express BLyS receptors. Gelonin, a type 1 ribosome-inactivating toxin, lacks cell membrane binding domain and hence is nontoxic to intact cells. We generated a construct of recombinant gelonin (rGel) fused to BLyS to specifically target quiescent B-CLL lymphocytes. The construct rGel/BLyS specifically binds and internalizes through BAFF-R into CD19(+) B-CLL lymphocytes and induces apoptosis at nanomolar concentrations. In contrast, rGel alone was not able to internalize into these leukemic lymphocytes. Mechanistically, the rGel/BLyS construct inhibits protein synthesis with an IC(50) of less than 3 nM compared with more than 5000 nM for rGel toxin alone. This rGel/BLyS-mediated decrease in protein synthesis was associated with a decline in short-lived proteins such as MCL-1 and XIAP, the 2 survival proteins in B-CLL. There was a strong relationship between a decrease in these proteins and the cleavage of PARP, a hallmark feature of apoptosis. Taken together, these data suggest that the rGel/BLyS fusion toxin may have potential therapeutic efficacy for B-CLL patients. PMID- 17119118 TI - Arginase 1 is expressed in myelocytes/metamyelocytes and localized in gelatinase granules of human neutrophils. AB - Arginase 1 (ARG1) metabolizes arginine, thus reducing the availability of arginine as a substrate for nitric oxide synthase (NOS). The decreased production of nitric oxide (NO) by NOS and the production of ornithine by ARG1 affect immune responses and tissue regeneration at sites of infection, respectively. We here demonstrate that ARG1 is synthesized in myelocytes/metamyelocytes and is stored in gelatinase granules. In accordance with this, activated neutrophils coreleased ARG1 and gelatinase to the extracellular environment on stimulation with phorbol 12-myristate 13-acetate (PMA), formyl-methionyl-leucyl-phenylalanine (fMLP), or tumor necrosis factor alpha (TNF-alpha). Overall, these findings define ARG1 as a genuine gelatinase granule protein and support a model in which activated neutrophils release ARG1 at sites of infection to modulate immune responses and promote tissue regeneration. PMID- 17119119 TI - A neoepitope generated by an FLT3 internal tandem duplication (FLT3-ITD) is recognized by leukemia-reactive autologous CD8+ T cells. AB - The FLT3 receptor tyrosine kinase is expressed in more than 90% of acute myelogeneous leukemias (AMLs), up to 30% of which carry an internal tandem duplication (ITD) within the FLT3 gene. Although varying duplication sites exist, most FLT3-ITDs affect a single protein domain. We analyzed the FLT3-ITD of an AML patient for encoding HLA class I-restricted immunogenic peptides. One of the tested peptides (YVDFREYEYY) induced in vitro autologous T-cell responses restricted by HLA-A*0101 that were also detectable ex vivo. These peptide reactive T cells recognized targets transfected with the patient's FLT3-ITD, but not wild-type FLT3, and recognized the patient's AML cells. Our results demonstrate that AML leukemic blasts can in principle process and present immunogenic FLT3-ITD neoepitopes. Therefore, FLT3-ITD represents a potential candidate target antigen for the immunotherapy of AML. PMID- 17119120 TI - Physiologic and aberrant regulation of memory T-cell trafficking by the costimulatory molecule CD28. AB - Productive T-cell immunity requires both the activation and the migration of specific T cells to the antigenic tissue. The costimulatory molecule CD28 plays an essential role in the initiation of T-cell-mediated immunity. We investigated the possibility that CD28 may also regulate migration of primed T cells to target tissue. In vitro, CD28-mediated signals enhanced T-cell transendothelial migration, integrin clustering, and integrin-mediated migration. In vivo, T cells bearing a mutation in the CD28 cytoplasmic domain, which abrogates PI3K activation, displayed normal clonal expansion but defective localization to antigenic sites following antigenic rechallenge. Importantly, antibody-mediated CD28 stimulation led to unregulated memory T-cell migration to extra-lymphoid tissue, which occurred independently of T-cell receptor (TCR)-derived signals and homing-receptor expression. Finally, we provide evidence that CD28- and CTLA-4 mediated signals exert opposite effects on T-cell trafficking in vivo. These findings highlight a novel physiologic function of CD28 that has crucial implications for the therapeutic manipulation of this and other costimulatory molecules. PMID- 17119122 TI - TACI regulates IgA production by APRIL in collaboration with HSPG. AB - Transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI) is a member of the tumor necrosis factor (TNF) receptor family that serves as a receptor for B-cell activating factor of the TNF family (BAFF) and as a proliferation-inducing ligand (APRIL). Although TACI is reported to function as a positive or negative regulator for B-cell responses, its roles remain elusive. Experiments using TACI siRNA into B cells indicated that TACI positively regulated APRIL-induced IgA production in collaboration with heparan sulfate proteoglycans (HSPG). Furthermore, TACI negatively regulated BAFF-induced B-cell proliferation and production of IgA and IgG. In addition, B cells treated with heparitinase to denature HSPG showed that HSPG is essential for APRIL-induced B cell responses such as B-cell proliferation, IgG and IgA production, induction of activation-induced cytidine deaminase (AID), and noncanonical NF-kappaB2. In contrast, phosphorylation of physiological AID kinase, protein kinase A (PKA), was dependent on TACI. Importantly, coligation of TACI and HSPG by specific antibodies, but not by TACI or HSPG ligation itself, could induce the phosphorylation of PKA and IgA production instead of APRIL. Our findings indicate that simultaneous binding of TACI and HSPG on B cells with APRIL is crucial for IgA production. PMID- 17119121 TI - Retroviral vector insertion sites associated with dominant hematopoietic clones mark "stemness" pathways. AB - Evidence from model organisms and clinical trials reveals that the random insertion of retrovirus-based vectors in the genome of long-term repopulating hematopoietic cells may increase self-renewal or initiate malignant transformation. Clonal dominance of nonmalignant cells is a particularly interesting phenotype as it may be caused by the dysregulation of genes that affect self-renewal and competitive fitness. We have accumulated 280 retrovirus vector insertion sites (RVISs) from murine long-term studies resulting in benign or malignant clonal dominance. RVISs (22.5%) are located in or near (up to 100 kb [kilobase]) to known proto-oncogenes, 49.6% in signaling genes, and 27.9% in other or unknown genes. The resulting insertional dominance database (IDDb) shows substantial overlaps with the transcriptome of hematopoietic stem/progenitor cells and the retrovirus-tagged cancer gene database (RTCGD). RVISs preferentially marked genes with high expression in hematopoietic stem/progenitor cells, and Gene Ontology revealed an overrepresentation of genes associated with cell-cycle control, apoptosis signaling, and transcriptional regulation, including major "stemness" pathways. The IDDb forms a powerful resource for the identification of genes that stimulate or transform hematopoietic stem/progenitor cells and is an important reference for vector biosafety studies in human gene therapy. PMID- 17119123 TI - Mouse models of IgG- and IgM-mediated hemolysis. AB - Well-characterized mouse models of allo-immune antibody-mediated hemolysis would provide a valuable approach for gaining greater insight into the pathophysiology of hemolytic transfusion reactions. To this end, mouse red blood cells (mRBCs) from human glycophorin A transgenic (hGPA-Tg) donor mice were transfused into non Tg recipients that had been passively immunized with IgG or IgM hGPA-specific monoclonal antibodies (mAbs). In this novel murine "blood group system," mRBCs from hGPA-Tg mice are "antigen positive" and mRBCs from non-Tg mice are "antigen negative." Passive immunization of non-Tg mice with the IgG1 10F7 and IgG3 NaM10 2H12 anti-hGPA mAbs each induced rapid clearance of incompatible transfused hGPA Tg-mRBCs in a dose-response manner. Using various knockout mice as transfusion recipients, both the complement system and activating Fcgamma receptors were found to be important in the clearance of incompatible mRBCs by each of these IgG mAbs. In addition, the IgM E4 anti-hGPA mAb induced complement-dependent intravascular hemolysis of transfused incompatible hGPA-Tg-mRBCs accompanied by gross hemoglobinuria. These initial studies validate the relevance of these new mouse models for addressing important questions in the field of transfusion medicine. PMID- 17119124 TI - Gene expression profiles identify a role for cyclooxygenase 2-dependent prostanoid generation in BMP6-induced angiogenic responses. AB - The bone morphogenetic protein (BMP) family of proteins participates in regulation of angiogenesis in physiologic and pathologic conditions. To investigate the molecular mechanisms that contribute to BMP-dependent angiogenic signaling, we performed gene expression profiling of BMP6-treated mouse endothelial cells. We detected 77 mRNAs that were differentially regulated after BMP6 stimulation. Of these, cyclooxygenase 2 (Cox2) was among the most highly up regulated by BMP stimulation, suggesting a role for Cox2 as a downstream regulator of BMP-induced angiogenesis. Up-regulation of Cox2 by BMP6 was detected at both mRNA and protein levels in endothelial cells, and BMP6 increased production of prostaglandins in a Cox2-dependent fashion. BMP6 up-regulated Cox2 at the transcriptional level through upstream SMAD-binding sites in the Cox2 promoter. Pharmacologic inhibition of Cox2, but not Cox1, blocked BMP6-induced endothelial cell proliferation, migration, and network assembly. BMP6-dependent microvessel outgrowth was markedly attenuated in aortic rings from Cox2-/- mice or after pharmacologic inhibition of Cox2 in aortas from wild-type mice. These results support a necessary role for Cox2 in mediating proangiogenic activities of BMP6. These data indicate that Cox2 may serve as a unifying component downstream from disparate pathways to modulate angiogenic responses in diseases in which neovascularization plays an underlying pathophysiologic role. PMID- 17119125 TI - Development of a secondary autoimmune disorder after hematopoietic stem cell transplantation for autoimmune diseases: role of conditioning regimen used. AB - Patients undergoing autologous hematopoietic stem cell transplantation (auto HSCT) for autoimmune disease may have an added propensity to develop a second autoimmune disorder, given the genetic predisposition to autoimmunity. Therefore, we undertook a retrospective analysis of all patients who have undergone auto HSCT for an autoimmune disease in our institution to determine the occurrence of a second autoimmune disorder and possible risk factors. In all, 155 patients underwent auto-HSCT for various autoimmune diseases; of those patients, 6 manifested a distinct secondary autoimmune disease at a median of 8.5 months (range, 2-30 months) after auto-HSCT. There were 2 patients with systemic lupus erythematosus, conditioned with a regimen containing antithymocyte globulin (ATG), who developed factor VIII inhibitors with severe bleeding. There were 4 patients (2 with multiple sclerosis, one each with lupus and systemic sclerosis) who received an alemtuzumab-containing conditioning regimen who developed autoimmune cytopenias. Among the 155 patients, the frequency of secondary autoimmune complications was 16.0% with alemtuzumab (4/25), 1.9% for ATG (2/102), and 0% for conditioning regimens without lympho-depleting antibodies (0/28)-a difference that was found to be significantly higher with alemtuzumab exposure (P = .011). In contrast, sex, type of ATG used, and CD34-selection of peripheral blood stem cells were not found to be significantly associated with development of a secondary autoimmune disorder. PMID- 17119126 TI - Effect of von Willebrand factor Y/C1584 on in vivo protein level and function and interaction with ABO blood group. AB - Blood group O and the cysteine allele of the Y/C1584 change in von Willebrand factor (VWF) are enriched in type 1 VWD, but neither causes disease. We investigated the effect of C1584, alone and in combination with the ABO blood group, on the level and properties of plasma VWF. A cohort of 5052 blood donors was recruited: 50 donors were heterozygous for Y/C1584 and 5002 were homozygous for Y/Y1584. Mean VWF antigen (VWF:Ag) for heterozygotes (82 +/- 35 IUdL(-1)) was significantly lower than for homozygotes (111 +/- 37 IUdL(-1)) (P < .001). Foreach ABO blood group, VWF:Ag was decreased among Y/C1584 heterozygotes compared with Y/Y1584 homozygotes; a larger decrease was observed for group O. Among donors with VWF:Ag levels of 50 IUdL(-1) or lower, Y/C1584 heterozygosity was markedly enriched (18%) compared with the entire cohort (1.5%). Blood group O was enriched to a lesser extent (2.4%), but Y/C1584 in conjunction with group O was strikingly enriched (34.8%). VWF collagen binding activity (VWF:CB) and ristocetin cofactor activity (VWF:RCo) were significantly lower for Y/C1584 heterozygotes than for Y/Y1584 homozygotes, and a qualitative difference in Y/C1584 plasma VWF multimer profile was observed compared with that for Y/Y1584 VWF. The data support a multifactorial basis for low VWF levels in some individuals. PMID- 17119127 TI - Aberrant NF-kappaB signaling in lymphoma: mechanisms, consequences, and therapeutic implications. AB - The transcription factor NF-kappaB is a tightly regulated positive mediator of T- and B-cell development, proliferation, and survival. The controlled activity of NF-kappaB is required for the coordination of physiologic immune responses. However, constitutive NF-kappaB activation can promote continuous lymphocyte proliferation and survival and has recently been recognized as a critical pathogenetic factor in lymphoma. Various molecular events lead to deregulation of NF-kappaB signaling in Hodgkin disease and a variety of T- and B-cell non-Hodgkin lymphomas either up-stream or downstream of the central IkappaB kinase. These alterations are prerequisites for lymphoma cell cycling and blockage of apoptosis. This review provides an overview of the NF-kappaB pathway and discusses the mechanisms of NF-kappaB deregulation in distinct lymphoma entities with defined aberrant pathways: Hodgkin lymphoma (HL), diffuse large B-cell lymphoma (DLBCL), mucosa-associated lymphoid tissue (MALT) lymphoma, primary effusion lymphoma (PEL), and adult T-cell lymphoma/leukemia (ATL). In addition, we summarize recent data that validates the NF-kappaB signaling pathway as an attractive therapeutic target in T- and B-cell malignancies. PMID- 17119128 TI - The cover. Partridge Tureen. PMID- 17119129 TI - A piece of my mind. Capturing life. PMID- 17119130 TI - A world of pain: scientists explore factors controlling pain perception. PMID- 17119131 TI - Studies probe anesthesia, sleep links. PMID- 17119132 TI - 1918 flu vaccine effective in mice. PMID- 17119133 TI - Treatment of chronic insomnia with cognitive behavioral therapy vs zopiclone. PMID- 17119134 TI - Treatment of chronic insomnia with cognitive behavioral therapy vs zopiclone. PMID- 17119135 TI - Colorectal cancer risk following a negative colonoscopy. PMID- 17119136 TI - Colorectal cancer risk following a negative colonoscopy. PMID- 17119137 TI - Colonoscopic screening for colorectal cancer. PMID- 17119138 TI - Fluoxetine for treatment of anorexia nervosa. PMID- 17119139 TI - Consequences of the quality improvement revolution. PMID- 17119140 TI - Surgical vs nonoperative treatment for lumbar disk herniation: the Spine Patient Outcomes Research Trial (SPORT): a randomized trial. AB - CONTEXT: Lumbar diskectomy is the most common surgical procedure performed for back and leg symptoms in US patients, but the efficacy of the procedure relative to nonoperative care remains controversial. OBJECTIVE: To assess the efficacy of surgery for lumbar intervertebral disk herniation. DESIGN, SETTING, AND PATIENTS: The Spine Patient Outcomes Research Trial, a randomized clinical trial enrolling patients between March 2000 and November 2004 from 13 multidisciplinary spine clinics in 11 US states. Patients were 501 surgical candidates (mean age, 42 years; 42% women) with imaging-confirmed lumbar intervertebral disk herniation and persistent signs and symptoms of radiculopathy for at least 6 weeks. INTERVENTIONS: Standard open diskectomy vs nonoperative treatment individualized to the patient. MAIN OUTCOME MEASURES: Primary outcomes were changes from baseline for the Medical Outcomes Study 36-item Short-Form Health Survey bodily pain and physical function scales and the modified Oswestry Disability Index (American Academy of Orthopaedic Surgeons MODEMS version) at 6 weeks, 3 months, 6 months, and 1 and 2 years from enrollment. Secondary outcomes included sciatica severity as measured by the Sciatica Bothersomeness Index, satisfaction with symptoms, self-reported improvement, and employment status. RESULTS: Adherence to assigned treatment was limited: 50% of patients assigned to surgery received surgery within 3 months of enrollment, while 30% of those assigned to nonoperative treatment received surgery in the same period. Intent-to-treat analyses demonstrated substantial improvements for all primary and secondary outcomes in both treatment groups. Between-group differences in improvements were consistently in favor of surgery for all periods but were small and not statistically significant for the primary outcomes. CONCLUSIONS: Patients in both the surgery and the nonoperative treatment groups improved substantially over a 2 year period. Because of the large numbers of patients who crossed over in both directions, conclusions about the superiority or equivalence of the treatments are not warranted based on the intent-to-treat analysis. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00000410. PMID- 17119141 TI - Surgical vs nonoperative treatment for lumbar disk herniation: the Spine Patient Outcomes Research Trial (SPORT) observational cohort. AB - CONTEXT: For patients with lumbar disk herniation, the Spine Patient Outcomes Research Trial (SPORT) randomized trial intent-to-treat analysis showed small but not statistically significant differences in favor of diskectomy compared with usual care. However, the large numbers of patients who crossed over between assigned groups precluded any conclusions about the comparative effectiveness of operative therapy vs usual care. OBJECTIVE: To compare the treatment effects of diskectomy and usual care. DESIGN, SETTING, AND PATIENTS: Prospective observational cohort of surgical candidates with imaging-confirmed lumbar intervertebral disk herniation who were treated at 13 spine clinics in 11 US states and who met the SPORT eligibility criteria but declined randomization between March 2000 and March 2003. INTERVENTIONS: Standard open diskectomy vs usual nonoperative care. MAIN OUTCOME MEASURES: Changes from baseline in the Medical Outcomes Study Short-Form Health Survey (SF-36) bodily pain and physical function scales and the modified Oswestry Disability Index (American Academy of Orthopaedic Surgeons/MODEMS version). RESULTS: Of the 743 patients enrolled in the observational cohort, 528 patients received surgery and 191 received usual nonoperative care. At 3 months, patients who chose surgery had greater improvement in the primary outcome measures of bodily pain (mean change: surgery, 40.9 vs nonoperative care, 26.0; treatment effect, 14.8; 95% confidence interval, 10.8-18.9), physical function (mean change: surgery, 40.7 vs nonoperative care, 25.3; treatment effect, 15.4; 95% CI, 11.6-19.2), and Oswestry Disability Index (mean change: surgery, -36.1 vs nonoperative care, -20.9; treatment effect, 15.2; 95% CI, -18.5. to -11.8). These differences narrowed somewhat at 2 years: bodily pain (mean change: surgery, 42.6 vs nonoperative care, 32.4; treatment effect, 10.2; 95% CI, 5.9-14.5), physical function (mean change: surgery, 43.9 vs nonoperavtive care 31.9; treatment effect, 12.0; 95% CI; 7.9-16.1), and Oswestry Disability Index (mean change: surgery -37.6 vs nonoperative care -24.2; treatment effect, -13.4; 95% CI, -17.0 to -9.7). CONCLUSIONS: Patients with persistent sciatica from lumbar disk herniation improved in both operated and usual care groups. Those who chose operative intervention reported greater improvements than patients who elected nonoperative care. However, nonrandomized comparisons of self-reported outcomes are subject to potential confounding and must be interpreted cautiously. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00000410. PMID- 17119142 TI - Prevention of nosocomial infection in cardiac surgery by decontamination of the nasopharynx and oropharynx with chlorhexidine gluconate: a randomized controlled trial. AB - CONTEXT: Nosocomial infections are an important cause of morbidity and mortality after cardiac surgery. Decolonization of endogenous potential pathogenic microorganisms is important in the prevention of nosocomial infections. OBJECTIVE: To determine the efficacy of perioperative decontamination of the nasopharynx and oropharynx with 0.12% chlorhexidine gluconate for reduction of nosocomial infection after cardiac surgery. DESIGN, SETTING, AND PARTICIPANTS: A prospective, randomized, double-blind, placebo-controlled clinical trial conducted at the Onze Lieve Vrouwe Gasthuis, Amsterdam, the Netherlands, between August 1, 2003, and September 1, 2005. Of 991 patients older than 18 years undergoing elective cardiothoracic surgery during the study interval, 954 were eligible for analysis. INTERVENTION: Oropharyngeal rinse and nasal ointment containing either chlorhexidine gluconate or placebo. MAIN OUTCOME MEASURES: Incidence of nosocomial infection, in addition to the rate of Staphylococcus aureus nasal carriage and duration of hospital stay. RESULTS: The incidence of nosocomial infection in the chlorhexidine gluconate group and placebo group was 19.8% and 26.2%, respectively (absolute risk reduction [ARR], 6.4%; 95% confidence interval [CI], 1.1%-11.7%; P = .002). In particular, lower respiratory tract infections and deep surgical site infections were less common in the chlorhexidine gluconate group than in the placebo group (ARR, 6.5%; 95% CI, 2.3% 10.7%; P = .002; and 3.2%; 95% CI, 0.9%-5.5%; P = .002, respectively). For the prevention of 1 nosocomial infection, 16 patients needed to be treated with chlorhexidine gluconate. A significant reduction of 57.5% in S aureus nasal carriage was found in the chlorhexidine gluconate group compared with a reduction of 18.1% in the placebo group (P<.001). Total hospital stay for patients treated with chlorhexidine gluconate was 9.5 days compared with 10.3 days in the placebo group (ARR, 0.8 days; 95% CI, 0.24-1.88; P = .04). CONCLUSION: Decontamination of the nasopharynx and oropharynx with chlorhexidine gluconate appears to be an effective method to reduce nosocomial infection after cardiac surgery. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00272675. PMID- 17119143 TI - Public roles of US physicians: community participation, political involvement, and collective advocacy. AB - CONTEXT: Whether physicians have a professional responsibility to address health related issues beyond providing care to individual patients has been vigorously debated. Yet little is known about practicing physicians' attitudes about or the extent to which they participate in public roles, which we defined as community participation, political involvement, and collective advocacy. OBJECTIVES: To determine the importance physicians assign to public roles, their participation in related activities, and sociodemographic and practice factors related to physicians' rated levels of importance and activity. DESIGN, SETTING, AND PARTICIPANTS: Mail survey conducted between November 2003 and June 2004 of 1662 US physicians engaged in direct patient care selected from primary care specialties (family practice, internal medicine, pediatrics) and 3 non-primary care specialties (anesthesiology, general surgery, cardiology). MAIN OUTCOME MEASURES: Rated importance of community participation, political involvement, collective advocacy, and relevant self-reported activities encompassing the previous 3 years; rated importance of physician action on different issues. RESULTS: Community participation, political involvement, and collective advocacy were rated as important by more than 90% of respondents, and a majority rated community participation and collective advocacy as very important. Nutrition, immunization, substance abuse, and road safety issues were rated as very important by more physicians than were access-to-care issues, unemployment, or illiteracy. Two thirds of respondents had participated in at least 1 of the 3 types of activities in the previous 3 years. Factors independently related to high overall rating of importance (civic-mindedness) included age, female sex, underrepresented race/ethnicity, and graduation from a non-US or non-Canadian medical school. Civic mindedness, medical specialty, practice type, underrepresented race/ethnicity, preceptors of physicians in training, rural practice, and graduation from a non-US or non-Canadian medical school were independently related to civic activity. CONCLUSIONS: Public roles are definable entities that have widespread support among physicians. Civic-mindedness is associated primarily with sociodemographic factors, but civic action is associated with specialty and practice-based factors. PMID- 17119144 TI - Botulism in 4 adults following cosmetic injections with an unlicensed, highly concentrated botulinum preparation. AB - CONTEXT: Botulism is a potentially lethal paralytic disease caused primarily by toxins of the anaerobic, spore-forming bacterium Clostridium botulinum. Although botulinum toxin A is available by prescription for cosmetic and therapeutic use, no cases of botulism with detectable serum toxin have previously been attributed to cosmetic or therapeutic botulinum toxin injections. On November 27, 2004, 4 suspected botulism case-patients with a link to cosmetic botulinum toxin injections were reported to the Centers for Disease Control and Prevention. OBJECTIVE: To investigate the clinical, epidemiological, and laboratory aspects of 4 suspected cases of iatrogenic botulism. DESIGN, SETTING, AND PATIENTS: Case series on 4 botulism case-patients. MAIN OUTCOME MEASURES: Clinical characteristics of the 4 case-patients, epidemiological associations, and mouse bioassay neutralization test results from case-patient specimens and a toxin sample. RESULTS: Clinical characteristics of the 4 case-patients were consistent with those of naturally occurring botulism. All case-patients had been injected with a highly concentrated, unlicensed preparation of botulinum toxin A and may have received doses 2857 times the estimated human lethal dose by injection. Pretreatment serum toxin levels in 3 of the 4 case-patients were equivalent to 21 to 43 times the estimated human lethal dose; pretreatment serum from the fourth epidemiologically linked case-patient was not available. A 100-microg vial of toxin taken from the same manufacturer's lot as toxin administered to the case patients contained a toxin amount sufficient to kill approximately 14,286 adults by injection if disseminated evenly. CONCLUSIONS: These laboratory-confirmed cases of botulism demonstrate that clinical use of unlicensed botulinum toxin A can result in severe, life-threatening illness. Further education and regulation are needed to prevent the inappropriate marketing, sale, and clinical use of unlicensed botulinum toxin products. PMID- 17119145 TI - Potential for genetics to promote public health: genetics research on smoking suggests caution about expectations. PMID- 17119146 TI - Interpreting surgical trials with subjective outcomes: avoiding UnSPORTsmanlike conduct. PMID- 17119147 TI - Surgical treatment of lumbar disk disorders. PMID- 17119148 TI - JAMA patient page. Herniated lumbar disks. PMID- 17119149 TI - Free access for all! Can we afford it? PMID- 17119150 TI - Altered cardiac myocyte Ca regulation in heart failure. AB - Ca is critical in both the electrical and mechanical properties of cardiac myocytes, and much is known about ionic currents and the normal excitation contraction coupling process. In heart failure, there are significant alterations in how myocyte Ca is regulated, and these alterations are critical in dictating both contractile dysfunction and certain cardiac arrhythmias that are characteristic of heart failure. PMID- 17119151 TI - What determines blood vessel structure? Genetic prespecification vs. hemodynamics. AB - Vascular network remodeling, angiogenesis, and arteriogenesis play an important role in the pathophysiology of ischemic cardiovascular diseases and cancer. Based on recent studies of vascular network development in the embryo, several novel aspects to angiogenesis have been identified as crucial to generate a functional vascular network. These aspects include specification of arterial and venous identity in vessels and network patterning. In early embryogenesis, vessel identity and positioning are genetically hardwired and involve neural guidance genes expressed in the vascular system. We demonstrated that, during later stages of embryogenesis, blood flow plays a crucial role in regulating vessel identity and network remodeling. The flow-evoked remodeling process is dynamic and involves a high degree of vessel plasticity. The open question in the field is how genetically predetermined processes in vessel identity and patterning balance with the contribution of blood flow in shaping a functional vascular architecture. Although blood flow is essential, it remains unclear to what extent flow is able to act on the developing cardiovascular system. There is significant evidence that mechanical forces created by flowing blood are biologically active within the embryo and that the level of mechanical forces and the type of flow patterns present in the embryo are able to affect gene expression. Here, we highlight the pivotal role for blood flow and physical forces in shaping the cardiovascular system. PMID- 17119152 TI - Pulmonary emphysema: when more is less. AB - Pulmonary emphysema results from the loss of intricate alveolar architecture and progressive simplification of small and highly effective gas-exchanging units into large, inefficient cyst-like spaces. Because of the loss of alveolar gas exchanging units and the capillary bed within them, blood oxygen levels eventually fall and pressures within the pulmonary circulation rise. Recent insights from genetically manipulated mouse models have refined our understanding of the molecular events that prevent or promote the development of pulmonary emphysema. Capitalizing on an improved molecular understanding of emphysema with improved therapeutics has the potential to enhance both the survival and quality of life of patients with this common lung disease. PMID- 17119153 TI - SIRT1: linking adaptive cellular responses to aging-associated changes in organismal physiology. AB - Sirtuins comprise a family of enzymes implicated in the determination of organismal life span in yeast and the nematode. The mammalian sirtuin SIRT1 has been shown to deacetylate several proteins in an NAD(+)-dependent manner. SIRT1 substrates are involved in the regulation of apoptosis/cell survival, endocrine signaling, differentiation, chromatin remodeling, and transcription. Thus SIRT1 provides a molecular link between nutrient availability and adaptive transcriptional responses. This review presents current evidence as to how SIRT1 functions are relevant to changes in tissue physiology that occur with ageing and its implications for future pharmacological intervention to alleviate such degenerative processes. PMID- 17119154 TI - Peptides targeting protein kinases: strategies and implications. AB - Protein kinases are important key regulators in most, if not all, biological processes and are linked with many human diseases. Protein kinases thus became attractive targets for drug design. Intracellularly active peptides that selectively interfere with kinase function and or kinase-mediated signaling pathways are potential drug compounds with therapeutic implications. PMID- 17119155 TI - The Amt/MEP/Rh family: structure of AmtB and the mechanism of ammonia gas conduction. AB - The atomic structures of the first members of the Amt/MEP/Rh family show that they are 11-crossing membrane proteins that form trimers in the membrane. Each monomer supports a hydrophobic channel that conducts NH(3) but not any water or ions. The reprotonation of NH(3) on the receiving side raises the pH on that side in the absence of metabolism of NH(3), and there is no transfer of protons through the protein. PMID- 17119156 TI - Lipid rafts, detergent-resistant membranes, and raft targeting signals. AB - Lipid rafts are liquid-ordered (l(o)) phase microdomains proposed to exist in biological membranes. Rafts have been widely studied by isolating l(o)-phase detergent-resistant membranes (DRMs) from cells. Recent findings have shown that DRMs are not the same as preexisting rafts, prompting a major revision of the raft model. Nevertheless, raft-targeting signals identified by DRM analysis are often required for protein function, implicating rafts in a variety of cell processes. PMID- 17119157 TI - PTEN regulation, a novel function for the p85 subunit of phosphoinositide 3 kinase. AB - Timely regulation of phosphatidylinositol-3,4-bisphosphate [PI(3,4)P2] and phosphatidylinositol-3,4,5-trisphosphate [PI(3,4,5)P3] abundance in cells is essential for the control of cellular homeostasis. The concentrations of these lipids are low in quiescent cells but rapidly and transiently increase following growth factor receptor (GFR) stimulation, which triggers cellular metabolic changes, proliferation, survival, and motility. Class I(A) phosphatidylinositol 3 kinase (PI3K), which is composed of a p85 (regulatory) and p110 (catalytic) subunits, is the enzyme generating PI(3,4)P2 and PI(3,4,5)P3 following GFR stimulation. Although the steps in GFR-induced activation of PI3K , are relatively well known, the mechanisms for subsequent 3-polyphospho-PI down regulation are less understood. Examination of frequent genetic alterations in human cancer showed that PTEN (phosphatase with tensin homology on chromosome 10) is the major enzyme that decreases PI(3,4)P2 and PI(3,4,5)P3 cell content. Nonetheless, interpretation of the complexity of PTEN regulation remains a matter of debate. The recent description of diminished PTEN activity in liver conditional knockout mice lacking the p85alpha PI3K regulatory subunit reveals a previously unknown p85alpha-dependent negative-feedback pathway that controls PI(3,4)P2 and PI(3,4,5)P3 half-life by regulating PTEN. PMID- 17119158 TI - Ubiquitin and NEDD8: brothers in arms. AB - Ubiquitin (Ub) and ubiquitin-like (Ubl) proteins are small signaling molecules that are involved in many aspects of cell function. It has been assumed that Ub and Ubl have functionally distinct roles because they use different conjugation machineries and bind to different effector proteins. This paradigm, however, must be revisited after recent findings that signaling cascades mediated by Ub and the Ubl NEDD8 (Neural precursor cell-Expressed Developmentally Down-regulated 8) in the regulation of epidermal growth factor receptor (EGFR) endocytosis are redundant. In this context, Ub and NEDD8 share the same E3 ligase, Cbl, and are recognized by identical components of the endocytic sorting machinery. This unexpected redundancy introduces additional complexity to the current view of Ub signaling pathways. PMID- 17119159 TI - The clinician's responsibility for the consumer's financial well-being. PMID- 17119160 TI - Home nutrition support from the patient's perspective: the real reality story! PMID- 17119161 TI - Technology dependence in home care: impact on patients and their family caregivers. AB - BACKGROUND: The purpose of this review is to explore how home technology care affects patients, family caregivers, and quality of life (QOL). METHODS: A literature search was conducted to identify studies of home parenteral nutrition (HPN) and other technology prescribed home care. RESULTS: Technology dependence influences health-related QOL. Patients and their family caregivers must balance the positive aspects of being in the home environment with the challenges of administering complex therapies at home. Patients and caregivers need additional support to reduce the physical, emotional, social, and financial burdens they experience. CONCLUSIONS: More research is needed to address effective interventions to reduce patient and caregiver burdens and to improve outcomes for technology-dependent individuals. A greater level of preparedness for managing home technology and technology-related problems may improve quality of life. PMID- 17119162 TI - Fostering coping skills and resilience in Home Enteral Nutrition (HEN) consumers. AB - BACKGROUND: Home enteral nutrition (HEN) is a lifesaving therapy that provides benefits along with countless challenges. This qualitative study examined how HEN consumers learned to cope successfully with HEN-related challenges and uncovered how healthcare providers could help foster the process of coping in other HEN consumers. METHODS: Twelve adult HEN consumers who perceived that they were coping successfully and overcoming the adversity associated with HEN, and met the criteria for resilience using the Resilience Scale, self-selected for the study. Participants engaged in a series of 2 in-depth interviews. Data were coded and analyzed using grounded theory methodology. RESULTS: One overarching theme and 5 main categories emerged from the data, revealing that these individuals coped successfully with HEN by developing an attitude of personal responsibility to accept new life conditions, take charge of their own well-being, seek and accept support, maximize independence and normality, and focus on the positive. In addition, these participants used a variety of problem- and emotion-focused coping strategies and shared resilient characteristics such as self-efficacy and perseverance. CONCLUSIONS: Implications for clinical practice and HEN education, along with suggestions for healthcare providers to foster coping with HEN are provided. An educational manual with self-help suggestions for adult HEN consumers is also available at http://www.copingwell.com. PMID- 17119163 TI - Micronutrient assessment in long-term home parenteral nutrition patients. AB - Assessment of micronutrient status of patients requiring long-term parenteral nutrition (PN) continues to be a challenge for clinicians. The scientific literature primarily consists of small studies and case reports. There are no evidence-based guidelines available. Clinicians rely on clinical assessment according to the patient's disease process, medication regimen, physical examination, and history of nutrient intake to determine existence or risk of micronutrient deficiency and toxicity. This paper will examine current scientific evidence and existing recommendations for the micronutrient assessment and management of patients requiring long-term PN. PMID- 17119164 TI - Reinstituting oral feeding in tube-fed adult patients with dysphagia. AB - Feeding tubes are valuable assets in the rehabilitation of adult patients with dysphagia. Feeding tubes may be placed in response to perceived risks of airway compromise or insufficient nutrient intake. However, not all patients require long-term enteral feeding. With intensive dysphagia therapy, many patients will experience resolving deficits or improvement in swallowing ability. These patients require an appropriate strategy to transition from tube to oral feeding. This article reviews some of the basic characteristics of dysphagia and identifies specific swallowing difficulties in 2 groups of patients who often benefit from temporary enteral feeding: stroke survivors and patients treated for head and neck cancer. Specific suggestions are offered for clinical strategies to reinstitute oral feeding in these groups of tube-fed patients. PMID- 17119165 TI - Nutrition support and the chronic critical illness syndrome. AB - Critical illness can be viewed as consisting of 4 distinct stages: (1) acute critical illness (ACI), (2) prolonged acute critical illness, (3) chronic critical illness, and (4) recovery. ACI represents the evolutionarily programmed response to a stressor. In ACI, substrate is shunted away from anabolism and toward vital organ support and inflammatory proteins. Nutrition support in this stage is unproven and may ultimately prove detrimental. As critical illness progresses, there is no evolutionary precedent, and man owes his life to modern critical care medicine. It is at this point that nutrition and metabolic support become integral to the care of the patient. This paper (1) delineates and develops the 4 stages of critical illness using current evidence, clinical experience, and new hypotheses; (2) defines the chronic critical illness syndrome (CCIS); and (3) details an approach to the metabolic and nutrition support of the chronically critically ill patient using the metabolic model of critical illness as a guide. It is our hope that this clinical model can generate testable hypotheses that can improve the outcome of this unique population of patients. PMID- 17119166 TI - Intestinal failure-associated metabolic bone diseases and response to teriparatide. AB - Patients requiring home parenteral nutrition (PN) may develop metabolic bone disease, the etiology of which can be multifactorial. We report a case of significantly low bone mass in a postmenopausal woman with history of short bowel syndrome, renal impairment, and previous radiation exposure who responded to intermittent subcutaneous administration of parathyroid hormone. Her bone mineral density normalized after she completed a course of 18 months of treatment, and a bone isotope scan was negative for skeletal malignancy. PMID- 17119167 TI - A microbiologic study of enteral feeding hang time in a burn hospital: can feeding costs be reduced without compromising patient safety? AB - BACKGROUND: Procedural changes for hospitalized patients must always balance safety with fiscal constraints. Microbiologic contamination of enteral feeding solutions has been previously associated with nosocomial infections. Formula manipulation and hang time contribute to microbial load, and there is considerable variation in hang time recommendations in the medical literature. With cost containment in mind, the purpose of this performance improvement study was to determine if an increase in hang time of a modular tube feeding product would increase microbial load or affect the nosocomial infection rate in pediatric burn patients. METHODS: This biphasic trial initially evaluated the microbial load of the feeding after delivery of two 4-hour aliquots into a container using the same delivery set (total hang time of 8 hours; number of tests = 20). Second, once this feeding procedure was deemed microbiologically safe, tube feedings were administered to patients, and both microbial load and nosocomial infection rate were monitored for 1 year. RESULTS: Contamination levels at the end of the 8-hour period using the same feeding set with 2 consecutive 4-hour feeding aliquots (number of tests = 38) were lower than standard recommendations. The hospital's nosocomial infection rate was not altered by this procedural change, and feeding-set expenses were reduced. CONCLUSIONS: The hang time of our enteral feeding administration set can be increased safely from 4 hours to 8 hours, with the tube feeding preparation added as two 4-hour aliquots without a significant change in microbial load or nosocomial infection rate, thus promoting simultaneous fiscal responsibility and patient safety. PMID- 17119168 TI - Hypocaloric feeding of the critically ill. AB - During critical illness, the stress response causes accelerated gluconeogenesis and lipolysis, leading to hyperglycemia and elevated serum triglyceride levels. The traditional nutrition support strategy of meeting or exceeding calorie requirements may compound the metabolic alterations of the stress response. Hypocaloric nutrition support has the potential to provide nutrition support without exacerbating the stress response. Studies have shown hypocaloric nutrition support to be safe and to achieve nitrogen balance comparable with traditional regimens. Benefits shown include improved glycemic control, decreased intensive care unit (ICU) length of stay (LOS), and decreased ventilator days and infection rate; however, not all studies have produced identical results. Providing adequate dietary protein has emerged as an important factor in efficacy of the hypocaloric regimen. Although it is inconclusive, currently available research suggests that a nutrition support goal of 10-20 kcal/kg of ideal or adjusted weight and 1.5-2 g/kg ideal weight of protein may be beneficial during the acute stress response. Well-designed, randomized, controlled studies with adequate sample size that evaluate relevant clinical outcomes such as mortality, ICU LOS, and infection while controlling for factors such as glycemic control, severity of illness, incorporation of calories from all sources, in addition to feeding regimens, are needed to definitively determine the effects of hypocaloric nutrition support. PMID- 17119169 TI - Biography: Henry T. Randall, MD. PMID- 17119170 TI - The history of surgically placed feeding tubes. AB - Although supplemental enteral nutrition may have first been delivered by enema, the modern era of surgically placed feeding tubes began in the mid to late 1800s. Early procedures were generally disastrous, however, techniques rapidly improved. The basic techniques of surgical enteral access have not changed significantly in the last century, although endoscopic, radiologic and laparoscopic modifications have been described and adopted in the last 25 years. This article reviews some of the landmark surgical highlights in the United States and European literature regarding surgical enteral access. PMID- 17119172 TI - Short-chain fatty acids: ready for prime time? PMID- 17119173 TI - Idiopathic pericarditis. PMID- 17119174 TI - New skills acquisition. PMID- 17119175 TI - Nothing up the sleeve? PMID- 17119176 TI - Program director satisfaction with leadership skills. AB - CONTEXT: Educators in radiologic technology help students mature into health care professionals; however, not all educators receive formal training in education. The majority of radiologic technology educators train first as practitioners and later become educators. OBJECTIVE: Identify areas for professional development and effective methods to learn leadership skills. METHODS: Radiologic technology program directors from Joint Review Committee on Education in Radiologic Technology (JRCERT) accredited programs were surveyed and interviewed to determine their level of satisfaction with their leadership skills. RESULTS: Results indicate program directors are satisfied with most of their leadership skills. Budget and resources, conflict resolution and communication were identified as areas for improvement. Program directors identified workshops and lectures as their preferred method of learning leadership skills. SUMMARY: Program directors need to use a variety of skills for the successful operation of their programs. Even though results from this study indicate that radiologic technology program directors were satisfied overall with their leadership skills, there were areas that could use improvement. PMID- 17119177 TI - Occupational stress and radiography. AB - CONTEXT: Stress is a very common problem. There has been considerable research on stress in certain health care professions; however, very little is known about the importance of occupational stress on radiographers. The studies that do exist indicate that the prevalence and impact of stress on radiographers is considerable and that occupational stress in the radiography field is a very common health problem. OBJECTIVE: The aim of this article is to review the available evidence to assess the effect of occupational stress on radiographers and compare it to health care workers. METHODS: Several major databases were searched using various keywords. Additionally, references from selected articles were used to retrieve other papers relevant to the topic. RESULTS: A review of the literature revealed significant stress-related consequences on the physical and mental health of health care workers. These findings can be applied to radiographers and used to reduce stress in the workplace. SUMMARY: Future research in the area of health promotion, including interventional relaxation and stress relief courses, among radiographers is needed. PMID- 17119178 TI - Accreditation: The hallmark of educational quality. AB - CONTEXT: Although accreditation issues have received attention over time, an accurate examination of accreditation has been absent within the radiologic science literature. OBJECTIVE: Using a literature review format, this review will define accreditation, provide a historical review of accreditation and purposes, identify the types of accreditation, discuss the major issues related to accreditation and explain how accreditation has affected the radiologic sciences. RESULTS: Although there is a great deal of rhetoric about accreditation in the radiologic sciences, research to support a particular type of accreditation within the radiologic sciences is absent. SUMMARY: Accreditation programs continuously are reviewed and changed, and accreditation organizations constantly strive to improve quality. In the past, the radiologic sciences have been influenced by accreditation decisions. The potential for future accreditation decisions affecting the radiologic sciences profession is possible. PMID- 17119179 TI - Management concepts. AB - Management concepts evolve through time. Health care managers can learn new concepts by evaluating classical management strategies, as well as modern-day strategies. Focusing on quality improvement and team building can help managers align the goals of their departments with the goals of the organization, consequently improving patient care. PMID- 17119180 TI - Diagnostic imaging of traumatic brain injury. AB - In this Directed Reading, the history and epidemiology of traumatic brain injury (TBI) will be briefly introduced, the physical and physiological nature of TBI reviewed and the role of imaging in the assessment of TBI patients described. New imaging techniques and recent findings about the neurological correlates of TBI symptoms and outcomes from studies using different imaging modalities and techniques will also be discussed. This directed reading will focus on closed head TBI; penetrating missile brain injuries, such as those caused by bullet wounds, will not be reviewed. PMID- 17119181 TI - The CASL Study: Dose reduction in a computed radiography environment. PMID- 17119182 TI - Placenta accreta. PMID- 17119183 TI - Sinus series. PMID- 17119184 TI - Cutaneous community-associated methicillin-resistant staphylococcus aureus among all skin and soft-tissue infections in two geographically distant pediatric emergency departments. AB - OBJECTIVES: To describe the culture results of cutaneous infections affecting otherwise healthy children presenting to two pediatric emergency departments (EDs) in the southeastern United States and southern California. METHODS: Medical records of 920 children who presented to the pediatric EDs with skin infections and abscesses (International Classification of Diseases, Ninth Revision codes 680.0-686.9) during 2003 were reviewed. Chronically ill children with previously described risk factors for community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) were excluded. Data abstracted included the type of infection; the site of infection; and, if a culture was obtained, the organism grown, along with their corresponding sensitivities. RESULTS: Of the 270 children who had bacterial cultures obtained, 60 (22%) were CA-MRSA-positive cultures, most cultured from abscesses (80%). Of all abscesses cultured, CA-MRSA grew in more than half (53%). All CA-MRSA isolates tested were sensitive to vancomycin, trimethoprim-sulfamethoxazole, rifampin, and gentamicin. One isolate at each center was resistant to clindamycin. The sensitivities at both institutions were similar. CONCLUSIONS: The authors conclude that CA-MRSA is responsible for most abscesses and that the pattern of CA-MRSA infections in these geographically distant pediatric EDs is similar. These data suggest that optimal diagnostic and management strategies for CA-MRSA will likely be widely applicable if results from a larger, more collaborative study yield similar findings. PMID- 17119185 TI - Clinical features of bacterial conjunctivitis in children. AB - OBJECTIVES: Conjunctivitis is a common cause of primary care and emergency department (ED) visits. There is a paucity of data in recent literature on the prevalence of pediatric bacterial conjunctivitis, and there are no evidence-based clinical guidelines for empirical treatment. The study objective was to describe clinical features most predictive of bacterial conjunctivitis. METHODS: This was a prospective study in a children's hospital ED. Conjunctival swabs for bacterial culture were obtained from patients aged 1 month to 18 years presenting with red or pink eye and/or the diagnosis of conjunctivitis. RESULTS: A total of 111 patients were enrolled over one year. Patients had a mean (+/-SD) age of 33.2 (+/ 37.5) months, and 55% were male. Eighty-seven patients (78%) had positive bacterial cultures. Nontypeable Haemophilus influenzae accounted for 82% (71/87), Streptococcus pneumoniae for 16% (14/87), and Staphylococcus aureus for 2.2% (2/87). Five clinical variables were significantly associated with a positive bacterial culture. Regression analysis revealed that the combination of a history of gluey or sticky eyelids and the physical finding of mucoid or purulent discharge had a posttest probability of 96% (95% confidence interval = 90% to 99%). Subjective scoring by physicians for a positive culture was 50.6%. CONCLUSIONS: Conjunctivitis in children is predominantly bacterial, with nontypeable H. influenzae being the most common organism. A history of gluey or sticky eyelids and physical findings of mucoid or purulent discharge are highly predictive of bacterial infection. Based on the above data, empirical ophthalmic antibiotic therapy may be appropriate in children presenting with conjunctivitis. PMID- 17119186 TI - Emergency medicine practitioner knowledge and use of decision rules for the evaluation of patients with suspected pulmonary embolism: variations by practice setting and training level. AB - BACKGROUND: Several clinical decision rules (CDRs) have been validated for pretest probability assessment of pulmonary embolism (PE), but the authors are unaware of any data quantifying and characterizing their use in emergency departments. OBJECTIVES: To characterize clinicians' knowledge of and attitudes toward two commonly used CDRs for PE. METHODS: By using a modified Delphi approach, the authors developed a two-page paper survey including 15 multiple choice questions. The questions were designed to determine the respondents' familiarity, frequency of use, and comprehension of the Canadian and Charlotte rules. The survey also queried the frequency of use of unstructured (gestalt) pretest probability assessment and reasons why physicians choose not to use decision rules. The surveys were sent to physicians, physician assistants, and medical students at 32 academic and community hospitals in the United States and the United Kingdom. RESULTS: Respondents included 555 clinicians; 443 (80%) work in academic practice, and 112 (20%) are community based. Significantly more academic practitioners (73%) than community practitioners (49%) indicated familiarity with at least one of the two decision rules. Among all respondents familiar with a rule, 50% reported using it in more than half of applicable cases. A significant number of these respondents could not correctly identify a key component of the rule (23% for the Charlotte rule and 43% for the Canadian rule). Fifty-seven percent of all respondents indicated use of gestalt rather than a decision rule in more than half of cases. CONCLUSIONS: Academic clinicians were more likely to report familiarity with either of these two specific decision rules. Only one half of all clinicians reporting familiarity with the rules use them in more than 50% of applicable cases. Spontaneous recall of the specific elements of the rules was low to moderate. Future work should consider clinical gestalt in the evaluation of patients with possible PE. PMID- 17119187 TI - Factors associated with hospital admission among emergency department patients with chronic obstructive pulmonary disease exacerbation. AB - OBJECTIVES To determine the patient factors associated with hospital admission among adults who present to the emergency department (ED) with acute exacerbations of chronic obstructive pulmonary disease (COPD) and to determine whether admissions were concordant with recommendations in the Global Initiative for Chronic Obstructive Lung Disease (GOLD) guidelines. METHODS: The authors performed a prospective multicenter cohort study involving 29 EDs in the United States and Canada. By using a standard protocol, consecutive ED patients with COPD exacerbation were interviewed, and their charts were reviewed. Predictors of admission were determined by multivariate logistic regression. RESULTS: Of 384 patients, 233 (61%; 95% confidence interval = 56% to 66%) were admitted. Multivariate analysis showed that a higher likelihood of admission was associated with older age, female gender, more pack-years of smoking, recent use of inhaled corticosteroid, self-reported activity limitation in the past 24 hours, higher respiratory rate at ED presentation, and a concomitant diagnosis of pneumonia. Patients who reported the ED as their usual site for problem COPD care, or who had mixed COPD and asthma, were less likely to be admitted. The authors confirmed five of the seven testable indications for hospital admission in the GOLD guidelines. CONCLUSIONS: Several patient factors were independently associated with hospital admission among ED patients with COPD exacerbations. Overall, concordance with admission recommendations in the GOLD guidelines was high. The authors also identified a few novel predictors of admission (female gender, ED as the usual site for problem COPD care, mixed diagnosis of COPD and asthma, recent use of inhaled corticosteroid) that require replication in future studies. PMID- 17119188 TI - Point-of-care urine trypsinogen testing for the diagnosis of pancreatitis. AB - OBJECTIVES: To assess a point-of-care (POC) urine trypsinogen (UT) test for the diagnosis of pancreatitis in the emergency department (ED). METHODS: This was a prospective cohort study of a convenience sample of patients presenting to the ED with abdominal pain or symptoms suggestive of pancreatitis. A 3-minute POC UT test (Actim Pancreatitis; Medix Biochemica, Kauniainen, Finland) was compared with plasma lipase and amylase measurements, imaging results when performed, and final discharge diagnoses. The criterion standard was a final discharge diagnosis of acute pancreatitis. RESULTS: Of 191 patients included in this study, 17 patients were diagnosed with either acute or acute-on-chronic pancreatitis. The sensitivity and specificity of UT for acute pancreatitis were, respectively, 100% (95% confidence interval [CI] = 77% to 100%) and 96% (95% CI = 92% to 98%). Seven of the 17 patients with pancreatitis (41%) had diagnostic findings on CT and positive UT tests but had nondiagnostic plasma lipase and amylase levels. CONCLUSIONS: A POC UT screening test for pancreatitis in the ED compared favorably with plasma lipase and amylase levels. Future studies should be performed to explore whether this test in the ED setting has better clinical utility than plasma lipase or amylase. PMID- 17119189 TI - Clinicopathological conference: case report--a case of anorexia and weak arm. AB - The authors present the case of a 49-year-old female who presented to the emergency department with a chief complaint of "not eating well." She was found to have a heart murmur, a focal neurological deficit, and large mitral valve vegetation. The patient was later diagnosed with acute Pseudomonal endocarditis with septic emboli to the brain, liver, spleen, and kidneys. A discussion of the patient presentation, diagnostic evaluation, and outcome are reviewed. PMID- 17119190 TI - Identification of carcinogenic agents and primary prevention of cancer. AB - During the annual Ramazzini Days, the Mayor of Carpi confers the Ramazzini Award on scientists deemed by the Collegium Ramazzini to have made outstanding contributions to furthering the aims of Bernardino Ramazzini in safeguarding public health. Dr. Lorenzo Tomatis was the Ramazzini Award recipient in 2005, and the presentation of the award was a highlight of the Symposium. The Ramazzini Lecture given by Dr. Tomatis follows. PMID- 17119191 TI - Children's environmental health research--highlights from the Columbia Center for Children's Environmental Health. AB - A growing body of evidence has been generated indicating that the fetus, infant, and young child are especially susceptible to environmental toxicants as diverse as polycyclic aromatic hydrocarbons (PAHs), pesticides, lead, mercury, polychlorinated biphenyls (PCBs), and environmental tobacco smoke (ETS). Exposures to these toxicants may be related to the increases in recent decades in childhood asthma, cancer, and developmental disability. The Columbia Center for Children's Environmental Health (CCCEH), located in New York City, has developed four cohorts around the world to elucidate the relationships between these exposures and childhood illness. This article summarizes the recent findings from the Center's projects in the context of current research in children's environmental health. PMID- 17119192 TI - Interpreting cancer trends. AB - The interpretation of cancer incidence trends is complicated by short-term random variation, artifactual fluctuations introduced by screening, changes in diagnosis or disease classification, completeness of reporting, and by the multiplicity of factors that may affect risk for specific cancer sites. We analyzed trends in 56 different cancer sites and subsites in the U.S. SEER registries in the period 1975-2002 using join-point analysis. The increase in cancer incidence for all sites combined that became evident with the inception of the SEER registries in the mid-1970s has abated since the early 1990s. Among the 15 most common cancer sites in men, sites with increasing incidence rates during the most recent time period include melanoma of the skin and cancers of the prostate, kidney and renal pelvis (kidney), and esophagus. Among women, incidence rates are increasing for leukemia, non-Hodgkin's lymphoma, melanoma, and cancers of the breast, thyroid, urinary bladder, and kidney. Incidence rates for all childhood cancers combined increased 0.6% per year from 1975 to 2002. Cancer mortality rates have decreased in the United States since 1991 in both men and in women; site-specific death rates have decreased in the most recent time period for 12 of the top 15 cancer sites in men and 9 of the top 15 cancer sites in women. Similar trends in cancer incidence and mortality have been reported in other industrialized countries. Possible reasons for these trends are discussed. PMID- 17119193 TI - The anatomy of the exposures that occurred around the World Trade Center site: 9/11 and beyond. AB - The attack on the World Trade Center (WTC) resulted in a new era of awareness on terrorism in the United States and the issues surrounding the potential for acute and/or long-term health outcomes caused by personal exposures to toxicants released during a terrorist event or an accident. The aftermath of the collapse yielded a situation usually not encountered in environmental health science: a large population's exposure to a previously uncharacterized complex mixture of airborne gases and particles, and re-suspendable particles (>2.5 microm in diameter). This led to a series of rapidly changing potential and actual exposure categories, both in space and time that were associated with the complex mixture of heterogeneous composition and character; e.g., very large particles mixed with much smaller amounts of fine particles, and gases released by uncontrolled combustion. The four categories of outdoor exposure that were encountered will be discussed over the period from September 11 until the fires ended on December 20, 2001. Further, the complex issue of indoor exposure to deposited dust will be highlighted from the beginning through the residual exposure issues being examined today (Category 5 period). The strength of the information on the initial WTC dust and smoke, and the smoke plumes from the fires and the continuing (permanent) gaps in our knowledge within the exposure sciences will be discussed, as well as our attempt to reconstruct exposure for various segments of the population in southern Manhattan and the surrounding areas. This all will be tied to lessons that must be considered in response to future events, natural or otherwise. PMID- 17119194 TI - The health watch case-control study of leukemia and benzene: the story so far. AB - A case-control study nested in the Health Watch cohort of petroleum industry workers, investigated whether the excess of lymphohematopoetic cancers, identified among male members of the Health Watch cohort, was associated with benzene exposure. Cases of non-Hodgkin's lymphoma (n = 31), multiple myeloma (n = 15), and leukemia (n = 33) were identified between 1981 and 1999. Cases were age matched to five controls. Exposure was retrospectively estimated for each occupational history using an algorithm in a relational database. Benzene exposure measurements, supplied by Australian petroleum companies, were used to estimate exposure for specific tasks. The tasks carried out within the job, the products handled, and the technology used, were identified from interviews with contemporary colleagues. More than half of the subjects started work after 1965 and had an average exposure period of 20 years. Exposure was low, 85% of the cumulative exposure estimates were <10 ppm years. Matched analyses showed that non-Hodgkin's lymphoma and multiple myeloma were not associated with benzene exposure. Leukemia risk, however, was significantly increased for the subjects with greater than 16 ppm years cumulative exposure, odds ratio (OR) 51.9 (5.6 477) or with greater than 0.8 ppm intensity of highest exposed job. Cumulative exposures were similar to those found in comparable studies. The inclusion of occasional high exposures, for example, as a result of spillages, reduced the ORs, when the exposure was treated as either a continuous or a categorical variable. Our data demonstrate a strong association between leukemia and modest benzene exposure. The choice of cut-point and reference group has a marked effect on the ORs, but does not change the overall conclusions. PMID- 17119195 TI - Benzene exposure and multiple myeloma: a detailed meta-analysis of benzene cohort studies. AB - Case reports and epidemiological studies of workers exposed to benzene have demonstrated associations with a number of lymphohematopoietic diseases, but the association with multiple myeloma (MM) has been less apparent. Data from all of the "benzene cohort studies" conducted to date have been selected and evaluated for inclusion in a meta-analysis. The analysis demonstrates a significant excess in the relative risk (RR) of MM in relation to benzene exposure. Pooling the data from seven cohort studies, a meta-analysis yields a statistically significant weighted RR estimate of 2.13 (95% CI = 1.31-3.46). In the analysis of cohort data, an understanding of the cohort follow-up period in relation to benzene exposure and RR of MM is important. Exposure-related RRs of disease decline after the median latency periods are exceeded, particularly when exposure has terminated decades earlier. The positive epidemiological evidence for benzene as a cause of MM is supported by biological plausibility for such an effect from benzene exposure. Studies of refinery workers are difficult to interpret in relation to benzene exposure and risk of MM, but are limited in the study design and analysis. Nonetheless, they provide some support for an association between refinery work and MM. PMID- 17119196 TI - Dangerous and cancer-causing properties of products and chemicals in the oil refining and petrochemical industries. Part XXX: Causal relationship between chronic myelogenous leukemia and benzene-containing solvents. AB - Benzene and benzene-containing products and solvents have long been associated with bone marrow toxicity. Both animal studies and human epidemiological studies have shown statistically significant increases of leukemia and other lymphohematopoietic cancers in workers exposed to benzene. The most common leukemia that has been associated with benzene exposure, also called benzene poisoning, is acute myelocytic leukemia (AML). A review of the epidemiological literature on workers exposed to benzene or benzene-containing solvents and products shows, without question, that this exposure is significantly related to other types of leukemia and lymphoma. In this article, we review the literature on the relationship between benzene exposure and chronic myelogenous leukemia (CML) and find that benzene and benzene-containing products are significantly related to morbidity and mortality from CML. PMID- 17119197 TI - Causal relationship between non-Hodgkin's lymphoma and exposure to benzene and benzene-containing solvents. AB - Non-Hodgkin's lymphoma (NHL) is a malignant neoplasm of the lymphatic system made up of mainly B cell lymphocytes. A large number of studies have shown significant associations between NHL and benzene or benzene-containing solvents and products. This article summarizes studies detailing these associations and indicates those that are significant. Based on an analysis of the literature and the weight of evidence from numerous studies, it is reasonable to conclude that exposure to benzene or to solvents or products containing benzene is causally related to NHL. PMID- 17119198 TI - Genetic polymorphism of toxicant-metabolizing enzymes and prognosis of Chinese workers with chronic benzene poisoning. AB - Workers with chronic benzene poisoning (CBP) sometimes have a white blood cell count (WBC) below 4 x 10(9)/L even after cessation of workplace exposure to benzene for years. In order to explore this phenomenon, 120 workers with CBP were divided into two groups depending on the WBC, the mean diagnostic age of CBP, benzene exposure duration, and body mass index (BMI). The proportion of genotypes of cytochrome P450 2E1 (CYP2E1), glutathione-S-transferase mu-1 (GSTM1), glutathione-S-transferase theta-1 (GSTT1), myeloperoxidase (MPO), and NAD(P)H, quinone oxidoreductase 1 (NQO1) were compared between workers with WBC <4 x 10(9)/L and those with WBC > or =4 x 10(9)/L. With methods of logistic regression, a risk model was set up to predict the prognosis of CBP workers. The results indicated that the BMI of workers with WBC <4 x 10(9)/L was lower than that of workers with WBC of > or =4 x 10(9)/L (21.40 +/- 2.76 versus 23.09 +/- 3.36, P = 0.01), and the logistic regression model suggested there was a 4.5-fold increased risk among workers carrying GSTT1 null genotype (95% CI= 1.13- 17.54) compared with workers with GSTT1 non-null genotype. Our findings suggest that benzene exposure duration, BMI, and GSTT1 genotype may impact prognosis of the CBP workers. PMID- 17119199 TI - Gene-environment interactions in cancer: do they exist? AB - Single nucleotide polymorphisms (SNPs) are extensively used in case-control studies of practically all cancer types. In addition to the pure genetic studies, gene-environment studies, which simultaneously consider environmental factors, have been increasingly conducted. All SNP studies aim at the identification of the role of inherited cancer susceptibility genes. However, being genetic markers, they are applicable only on heritable conditions, which is often a neglected fact. Based on the data on the heritability of cancer and the importance of environmental factors in cancer etiology, we discuss the likelihood of successful gene-environment studies. The available evidence is not conclusive, but it consistently points to a minor heritable etiology in cancer, which will hamper the success of SNP-based association studies. We use simulation techniques to examine which situations would favor the application of a gene-environment approach instead of the traditional environmental approach in case-control studies. The results show that well-chosen candidate gene with a relatively low allele frequency may improve the power to detect environmental determinants of a disease. However, this advantage is lost when the number of underlying genes increases. We are concerned about an indiscriminate use of genetic tools for cancers, which are mainly environmental in origin. The likelihood of success for SNP-based gene-environment studies increases if established environmental risk factors are tested on proven candidate genes. Enhancing the likelihood that the disease causation is genetic, for example, by selecting familial cases, may increase the power of the studies, and the rareness of those cases calls for collaborative networks. PMID- 17119200 TI - Manufactured uncertainty: protecting public health in the age of contested science and product defense. AB - The strategy of "manufacturing uncertainty" has been used with great success by polluters and manufacturers of dangerous products to oppose public health and environmental regulation. This strategy entails questioning the validity of scientific evidence on which the regulation is based. While this approach is most identified with the tobacco industry, it has been used by producers of asbestos, benzene, beryllium, chromium, diesel exhaust, lead, plastics, and other hazardous products to avoid environmental and occupational health regulation. It is also central to the debate on global warming. The approach is now so common that it is unusual for the science not to be challenged by an industry facing regulation. Manufacturing uncertainty has become a business in itself; numerous technical consulting firms provide a service often called "product defense" or "litigation support." As these names imply, the usual objective of these activities is not to generate knowledge to protect public health but to protect a corporation whose products are alleged to have toxic properties. Evidence in the scientific literature of the funding effect--the close correlation between the results of a study desired by a study's funder and the reported results of that study- suggests that the financial interest of a study's sponsors should be taken into account when considering the study's findings. Similarly, the interpretation of data by scientists with financial conflicts should be seen in this light. Manufacturing uncertainty is antithetical to the public health principle that decisions be made using the best evidence currently available. PMID- 17119201 TI - Misuse of genetic data in environmental epidemiology. AB - The implications of attributing health and diseases more to "nature" (genes) or "nurture" (the environment) have been debated for a long time. Although considerable advancements have been made both in theoretical clarification of concepts, and in the study of the origins of disease, there is still much confusion, for example, in the press and in the beliefs of the population. There is a large consensus, among scientists, that only a small fraction of diseases is due to genes in the usual meaning, that is, according to mendelian inheritance (say around 5% of all diseases), whereas the vast majority of cases are due to environmental exposures or to "gene-environment interactions." In this article I will briefly discuss a model for gene-environment interactions, and I will recall an important discussion that took place decades ago around the mistakes related to attributing diseases or other traits to inheritance. Finally, I will describe a specific example of potential misuse of genetic information. PMID- 17119202 TI - Collision of basic and applied approaches to risk assessment of thyroid toxicants. AB - Thyroid hormone (TH) is essential for normal brain development; therefore, any environmental chemical that interferes sufficiently with thyroid function, TH metabolism, or TH action may exert adverse effects on brain development. Important known differences in aspects of thyroid endocrinology between the fetus, infant, and adult allow us to identify age-dependent vulnerabilities to thyroid toxicants with some confidence. These differences include the size of the hormone pool stored in the thyroid gland at different ages as well as the age dependent sensitivity to mild TH insufficiency. Several recent studies that describe risk assessments of the environmental contaminant, ammonium perchlorate, provide good examples of conclusions based on the selective consideration of these known aspects of the thyroid system. Specifically, authors who consider age dependent differences in thyroid endocrinology suggest that safe levels of perchlorate should be set at relatively low levels (low parts per billion). In contrast, authors who do not consider these known age-dependent differences in thyroid endocrinology recommend safe levels of perchlorate at high (hundreds) parts per billion to parts per million. Emerging evidence indicates that a variety of high production volume chemicals can directly interact with the TH receptor. As testing paradigms are designed by regulatory agencies, these age dependent differences in thyroid endocrinology must be considered. PMID- 17119203 TI - Science and policy in risk assessments of chlorinated ethenes. AB - In this article the use of data obtained from standardized experimental methods, for example, as specified in OECD guidelines for the testing of chemicals, epidemiology data, and mechanism data obtained from nonstandardized experimental methods in carcinogen risk assessment is scrutinized using the most recent risk assessments made by International Agency for Research on Cancer (IARC), the MAK(MAK)-Kommission, World Health Organization (WHO), European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC), and American Conference of Governmental Industrial Hygienists (ACGIH) for the four chlorinated ethenes as examples. The analysis shows that there was little controversy among these risk assessors about the interpretation of standardized animal data. On the other hand, they differ in their interpretation of epidemiology data, in particular in their assessment of statistical significance including the use of meta-analyses, and in quality evaluation of studies initiated on the basis of a priori concerns for carcinogenicity. The selection of mechanism data for species extrapolation is diverse among these risk assessors. Furthermore, in some cases they refrain from transparently motivating significant claims about mechanisms of toxicity by avoiding to give (explicit) references to the sources of information forming the basis of these claims or conclusions. This practice is not according to the scientific standards that should be required of a risk assessment document, and it makes it difficult to follow the argumentation and consequently to scrutinize the scientific accuracy of the conclusions drawn. In this article it is concluded that in some of these risk assessment documents, the use of mechanism data is not according to the scientific standards that should be required. It is furthermore concluded that if the use of mechanism data in these documents are representative of risk assessments in general, then there is an urgent need for further development and implementation of quality criteria for the use of mechanism data in species extrapolation. PMID- 17119204 TI - Potential health effects of occupational chlorinated solvent exposure. AB - Based on toxicology, metabolism, animal studies, and human studies, occupational exposure to chlorinated aliphatic solvents (methanes, ethanes, and ethenes) has been associated with numerous adverse health effects, including central nervous system, reproductive, liver, and kidney toxicity, and carcinogenicity. However, many of these solvents remain in active, large-volume use. This article reviews the recent occupational epidemiology literature on the most widely used solvents, methylene chloride, chloroform, trichloroethylene, and tetrachloroethylene, and discusses other chlorinated aliphatics. The impact of studies to date has been lessened because of small study size, inability to control for confounding factors, particularly smoking and mixed occupational exposures, and the lack of evidence for a solid pathway from occupational exposure to biological evidence of exposure, to precursors of health effects, and to health effects. International differences in exposure limits may provide a "natural experiment" in the coming years if countries that have lowered exposure limits subsequently experience decreased adverse health effects among exposed workers. Such decreases could provide some evidence that higher levels of adverse health effects were associated with higher levels of solvent exposure. The definitive studies, which should be prospective biomarker studies incorporating body burden of solvents as well as markers of effect, remain to be done. PMID- 17119205 TI - Endocrine disruptors: challenges for environmental research in the 21st century. AB - During the past 10 years, there has been a worldwide decline in the use and human exposure to many chemicals, including pesticides and persistent organic pollutants (POPs). However, a new generation of chemicals that have endocrine disrupting (ED) potential have emerged. Their presence in the environment and concomitant levels in humans are prevalent, although the sources of these contemporary-use industrial chemicals are not entirely identified. They include the phthalates, alkylphenols, brominated diphenyl ethers, and perfluorinated organics (PFOCs). The alkylphenols, especially bisphenol A, are potent EDs. Levels vary by geography, race/ethnicity, age and gender, and human health effects are just beginning to be assessed. This article discusses the toxicology, human exposure, and potential health effects of EDs that are likely to be important in the 21st century. PMID- 17119206 TI - Risk assessment of endocrine disrupters: the role of toxicological studies. AB - Endocrine disrupters (ED) represent a good challenge for experimental toxicology. In order to deal with several critical points relevant to risk assessment: (a) ED may induce long-term effects upon exposure in susceptible developmental phases, including postnatal life up to puberty; thus, efforts are required to refine testing strategies, for example, by supporting the two-generation rodent study with a comprehensive in vitro/in vivo screening battery; (b) due to the regulatory role of endocrine homeostasis, mechanisms of endocrine disruption may impact on immune, neurobehavioral, and reproductive development, as well as on susceptibility to cancer; (c) the potential multiple exposure to ED with common targets through diet and/or living environment calls for the development of models to understand mechanisms of interactions and effects of mixtures; and (d) last but not least, ED may interact with a number of factors related to differential vulnerability of individuals or population subgroups, including the intake of nutrients or bioactive food components. Besides reducing the chance for noxious chemicals to enter our life, toxicological research on mechanisms may also lead to the definition of possible biomarkers of exposure, effect, and susceptibility that may be further exploited in human health surveillance. PMID- 17119207 TI - Diesel exhaust and coal mine dust: lung cancer risk in occupational settings. AB - Conflicting evidence on the carcinogenicity of diesel exhaust (DE) and coal mine dust in occupational settings exist. Exposure measurement in most studies is inferred on the basis of job classifications and may lead to misclassification. Confounding behavioral factors (i.e., smoking) and occupational risk factors (exposure to asbestos, arsenic, radon) need to be considered. We evaluated the epidemiological evidence and current findings of the carcinogenicity of DE and coal mine dust in occupational settings. Pertaining literature was identified through Medline search and recent review articles. Strengths and limitations of recent approaches are discussed. Many epidemiological studies have addressed the question of carcinogenicity in workers exposed to DE, and most showed a low-to medium increase in the risk of bronchial carcinoma. The pooled relative risk (RR) estimates lie between 1.33 and 1.47, and a consistent rise in risk across various job categories and study designs point to a causal relationship. Data on the carcinogenicity of coal mine dust are less consistent and the potential for confounding by unmeasured risk factors (arsenic, radon, DE) are higher. While silica as one of its components has been evaluated as carcinogenic, there is inadequate evidence for the carcinogenicity of pure coal dust according to the International Agency for Research on Cancer (IARC). There is sufficient evidence for a causal relationship between DE and lung cancer in occupational settings. The evidence for coal mine dust is less convincing, but individual studies show an increase in risk of lung cancer in exposed workers. PMID- 17119208 TI - Lung tumor risk estimates from rat studies with not specifically toxic granular dusts. AB - Since 1985 several carcinogenicity studies have been published about lung tumors in rats after exposure to respirable granular biodurable particles without known significant specific toxicity (abbreviation of this complex definition by the three letters GBP to substitute the former term inert dusts). During this time, the relevance of the carcinogenicity of GBP in rats was questioned, for example, because no lung tumors from GBP were found in hamsters and carcinogenicity in mice was questionable. However, the carcinogenesis and the tumor risk from quartz appear similar in men and rats, and the effects of GBP in rats appear not to differ, on principle, from that of quartz, but at a much higher dose level. We calculated the excess risk (ER) of GBP in rats from the final results of an instillation study with 16 GBP types in connection with results of inhalation experiments with carbon black, titanium dioxide, and diesel particles. Retained particle volume together with some indicator of particle size was identified as the best suitable dose metric and the dose-response relationships were analyzed on the basis of the multistage model. By relating the results to the available dose-response slopes after inhalation, ER for workplace-like exposure were calculated for three particle size classes and an exposure to 0.3 mg/m(3) (density 2-2.5 g/mL); mean diameter 1.8-4 microm (GBP-fine-large): ER 0.1%; 0.09 0.2 microm (GBP-fine-small): ER 0.2%; 0.01-0.03 microm (GBP-ultra-fine): ER 0.5%. PMID- 17119209 TI - Measurements of asbestos burden in tissues. AB - Asbestos inhaled into the lung is recognized as a potential causal agent for the development of diseases in man. The diseases induced by asbestos include lung cancer, fibrosis of the lung (asbestosis), and extrapulmonary tumors including mesothelioma (a tumor of the serosal membrane), as well as fibrosis and other changes in the pleura linings. The cause of these diseases can often be more specifically linked to asbestos exposure once tissue burden of asbestos is established. The asbestos burden in tissue can be defined as the number of asbestos bodies and/or the numbers and types of asbestos fibers found in the tissue. In either of these cases the quality of information is directly dependent on the preparative techniques and instrumentation used in the analysis. The present article will discuss the significance of findings of tissue burden based on both these variables. PMID- 17119210 TI - Asbestos ban in India: challenges ahead. AB - Rapidly industrializing India is described by the International Monetary Fund as a young, disciplined, and vibrant economy with a projected growth of 6.7% for 2005. The total workforce of 397 million has only 7% of workers employed in the organized sector with construction, where asbestos exposure is prevalent, employing 4.4%. The domestic production of asbestos declined from 20,111 tons in 1998-1999 to 14,340 tons in 2002-2003. The imports from Russia and Canada increased from 61,474 tons in 1997-1998 to 97,884 tons in 2001-2002. The production of asbestos cement products went up from 0.68 million tons in 1993 1994 to 1.38 million tons in 2002-2003. The asbestos industry has been delicensed since March 2003. The number of asbestos-based units stood at 32, with the western state of Maharashtra having the largest number. According to official figures, the industry employs 8000 workers. The occupational exposure standard is still 2 fibers/mL, worse still, mesothelioma is not recognized as an occupational disease. The latest cancer registry data have no information on mesothelioma. The health and safety legislation does not cover 93% of workers in the unorganized sector where asbestos exposures are extremely high. Workers remain uninformed and untrained in dealing with asbestos exposure. Enforcement agencies are not fully conscious of the risks of asbestos exposure. Industrial hygiene assessment is seldom carried out and pathologists do not receive training in identifying mesothelioma histopathologically. The lack of political will and powerful influence of the asbestos industry are pushing India toward a disaster of unimaginable proportion. PMID- 17119211 TI - Ionizing radiation and cardiovascular disease. AB - For more than 15 years the A-bomb survivor studies have shown increased noncancer mortality due to radiation exposures. The most prominent cause of this increase is circulatory disease mortality. Although the estimated relative risk is less than for solid cancers (1.2 versus 1.6 per Sv), there are measurable increases in cardiovascular disease mortality at doses greater than 0.5 Sv. The evidence for circulatory diseases in mortality studies of occupational cohorts exposed to external radiation is less compelling. It is generally accepted that atherosclerosis is an inflammatory disease of the arteries and a risk factor for myocardial infarction. Immunological markers for inflammatory disease have been shown to be dose related in A-bomb survivors. Evidence from animal studies reveals increased cardiovascular mortality and arterial endothelial damage from both neutron and, to a lesser extent, gamma exposures. PMID- 17119212 TI - Power-frequency electric and magnetic fields in the light of Draper et al. 2005. AB - Power-frequency electric and magnetic fields are produced wherever electricity is used; exposure is ubiquitous. Epidemiologic studies find an association between children living in homes with the highest magnetic fields and childhood leukemia, but bias is a possible alternative to a causal explanation. A new study, Draper et al., looks at residence close to high-voltage power lines, one source of exposure to such fields, and its design avoids any obvious bias. It finds elevated childhood leukemia rates, but extending too far from the power lines to be straightforwardly compatible with the existing literature. This leads to an examination of alternative explanations: magnetic fields, other physical factors, such as corona ions, the characteristics of the areas power lines pass through, bias, and chance. The conclusion is that there is currently no single preferred explanation, but that this is a serious body of science that needs further work until an explanation is found. PMID- 17119213 TI - Getting it right the first time: developing nanotechnology while protecting workers, public health, and the environment. AB - Nanotechnology, the design and manipulation of materials at the atomic scale, may well revolutionize many of the ways our society manufactures products, produces energy, and treats diseases. Innovative nanotechnology products are already reaching the market in a wide variety of consumer products. Some of the observed properties of nanomaterials call into question the adequacy of current methods for determining hazard and exposure, and for controlling resulting risks. Given the limitations of existing regulatory tools and policies, two distinct kinds of initiatives are urgently needed: first, a major increase in the federal investment nanomaterial risk research, and second, rapid development and implementation of voluntary standards of care pending development of adequate regulatory safeguards. The U.S. government should increase federal funding for nanomaterial risk research under the National Nanotechnology Initiative to at least $100 million annually for the next several years. Several voluntary programs are currently at various stages of evolution, though the eventual outputs of each of these are still far from clear. Ultimately, effective regulatory safeguards, harmonized globally, are necessary to provide a level playing field for industry while adequately protecting human health and the environment. PMID- 17119214 TI - Pesticides and adult respiratory outcomes in the agricultural health study. AB - In the 1700s, Bernardino Ramazzini was among the first to describe respiratory disease among agricultural workers. Since then, farmers continue to have higher rates of respiratory illnesses, even as changes occur in occupational and environmental exposures on farms. While grain and animal exposures have been well studied for their role in agricultural lung diseases, pesticide exposures have not. Using the Agricultural Health Study, a prospective cohort study of approximately 89,000 licensed pesticide applicators and their spouses in Iowa and North Carolina, we are currently assessing the association of pesticides with respiratory outcomes, including wheeze, adult asthma, farmer's lung, and chronic bronchitis. At enrollment (1993-1997), 19% of farmers and 22% of commercial pesticide applicators reported wheeze in the previous year. Using logistic regression models adjusted for age, state, smoking status, and body mass index, we evaluated the association of 40 individual pesticides with wheeze within these two groups separately. In both groups, we observed strong evidence of an association of organophosphates with wheeze. For farmers, the organophosphates chlorpyrifos, malathion, and parathion were positively associated with wheeze; for the commercial applicators, the organophosphates chlorpyrifos, dichlorvos, and phorate were positively associated with wheeze. Chlorpyrifos was strongly associated with wheeze in a dose-dependent manner in both groups; use of chlorpyrifos for at least 20 days per year had an odds ratio of 1.48 (95% confidence interval [CI] = 1.00-2.19) for farmers and 1.96 (95% CI = 1.05-3.66) for commercial applicators. Our wheeze results are consistent with recent animal models that support a role for organophosphates and respiratory outcomes. PMID- 17119215 TI - The assessment of occupational exposure to diazinon in Nicaraguan plantation workers using saliva biomonitoring. AB - A cross-sectional study with repeated sample collection in multiple days was conducted to assess diazinon exposures. Saliva and limited blood samples were collected from 10 banana plantation workers involved with diazinon application and their children aged 2-12 years living in Chinandega, Nicaragua. Diazinon concentration-time profiles in saliva varied between two plantations, which reflects the differences of work practices in each plantation. Salivary concentrations of diazinon measured in Plantation 1 applicators continued to increase 2 days after self-reported diazinon application, suggesting an ongoing exposure among these workers. However, salivary diazinon concentrations measured in Plantation 2 applicators were peaked 12 h prior to the first application, and then decreased 36 h post the first application. Diazinon concentrations in saliva were significantly correlated with the time-matched plasma samples collected from the same workers, which is in agreement with the previous published data from animal models. Children's exposure to diazinon through take-home pathway does not exist, as evident by the majority of nondetected saliva samples, and this finding was confirmed by the results from the urine samples. Severe dehydration was observed in many plantation workers and their children, resulting in the loss of some saliva samples, which no doubt have impaired the overall quality of the study results. Regardless, this article has demonstrated that saliva can be used to assess exposures to diazinon in pesticide applicators and children. PMID- 17119216 TI - Cancer and pesticides: an overview and some results of the Italian multicenter case-control study on hematolymphopoietic malignancies. AB - Exposure to pesticides is recognized as an important environmental risk factor associated with development of cancer. Epidemiological studies, although sometimes contradictory, have linked phenoxy acid herbicides with non-Hodgkin's lymphoma (NHL) and Soft Tissue Sarcoma (STS); organochlorine insecticides with STS, NHL, and leukemia; organophosphorous compounds with NHL and leukemia; and triazine herbicides with ovarian cancer. Exposure assessment is a crucial point in studying the association between cancer and pesticides. In order to investigate the association between hematolymphopoietic malignancies and occupational exposures, including pesticides, a population-based case-control study was carried out in Italy in 11 areas, 9 of which are agricultural or mixed areas. All newly diagnosed cases of hematolymphopoietic malignancies were collected in a 3-year period (1991-1993). The control group consisted of a random sample of the population residing in each area. The approach to infer exposures in agriculture was based on: the use of an agricultural questionnaire with 24 crop-specific questionnaires; expert agronomists who reviewed the collected information for each subject and translated it into pesticides histories. In total, 1925 cases and 1232 controls were interviewed in the nine agricultural areas. Increased risk was observed for some specific classes of pesticides. Furthermore, a nonstatistically significant increased risk of NHL was observed for subjects who were exposed to phenoxy herbicides not using protective equipment and a significant increased risk for exposure to 2, 4-dichlorophenoxy acetic acid (2,4-D). PMID- 17119217 TI - Geographic model and biomarker-derived measures of pesticide exposure and Parkinson's disease. AB - For more than two decades, reports have suggested that pesticides and herbicides may be an etiologic factor in idiopathic Parkinson's disease (PD). To date, no clear associations with any specific pesticide have been demonstrated from epidemiological studies perhaps, in part, because methods of reliably estimating exposures are lacking. We tested the validity of a Geographic Information Systems (GIS)-based exposure assessment model that estimates potential environmental exposures at residences from pesticide applications to agricultural crops based on California Pesticide Use Reports (PUR). Using lipid-adjusted dichlorodiphenyldichloroethylene (DDE) serum levels as the "gold standard" for pesticide exposure, we conducted a validation study in a sample taken from an ongoing, population-based case-control study of PD in Central California. Residential, occupational, and other risk factor data were collected for 22 cases and 24 controls from Kern county, California. Environmental GIS-PUR-based organochlorine (OC) estimates were derived for each subject and compared to lipid adjusted DDE serum levels. Relying on a linear regression model, we predicted log transformed lipid-adjusted DDE serum levels. GIS-PUR-derived OC measure, body mass index, age, gender, mixing and loading pesticides by hand, and using pesticides in the home, together explained 47% of the DDE serum level variance (adjusted r(2) = 0.47). The specificity of using our environmental GIS-PUR derived OC measures to identify those with high-serum DDE levels was reasonably good (87%). Our environmental GIS-PUR-based approach appears to provide a valid model for assessing residential exposures to agricultural pesticides. PMID- 17119218 TI - The construction industry. PMID- 17119219 TI - Frequency and quality of radiation monitoring of construction workers at two gaseous diffusion plants. AB - Construction workers were and are considered temporary workers at many construction sites. Since World War II, large numbers of construction workers were employed at U.S. Department of Energy nuclear weapons sites for periods ranging from a few days to over 30 years. These workers performed tasks during new construction and maintenance, repair, renovation, and demolition of existing facilities. Such tasks may involve emergency situations, and may entail opportunities for significant radiation exposures. This paper provides data from interviews with more than 750 construction workers at two gaseous diffusion plants (GDPs) at Paducah, Kentucky, and Portsmouth, Ohio regarding radiation monitoring practices. The aim was to determine the extent to which workers believed they were monitored during tasks involving potential radiation exposures. The adequacy of monitoring practices is important for two reasons: (a) Protecting workers from exposures: Construction workers were employed by sub contractors, and may frequently been excluded from safety and health programs provided to permanent employees; and (b) Supporting claims for compensation: The Energy Employees Occupational Illness Compensation Program Act (EEOICPA) requires dose reconstruction of radiation exposures for most workers who file a claim regarding cancer. The use of monitoring data for radiation to qualify a worker means that there should be valid and complete monitoring during the work time at the various nuclear plants or workers may be unfairly denied compensation. The worker interviews from Paducah and Portsmouth were considered especially useful because these sites were designated as Special Exposure Cohorts (SECs) and the workers did not have to have a dose reconstruction to qualify for compensation for most cancers. Therefore, their responses were less likely to be affected by compensation concerns. Interview questions included asking for information regarding whether monitoring was performed, how often, and the maintenance (calibration) of monitoring equipment (devices). PMID- 17119220 TI - Evaluation of exposure to PAHs in asphalt workers by environmental and biological monitoring. AB - In the present article we assessed exposure to polycyclic aromatic hydrocarbons (PAHs) in Italian asphalt workers (AW, n = 100), exposed to bitumen fumes and diesel exhausts, and in roadside construction workers (CW, n = 47), exposed to diesel exhausts, by means of environmental and biological monitoring. 1 hydroxypyrene (OH-Py) was determined in urine spot samples collected, respectively, after 2 days of vacation (baseline), before, and at the end of the monitored work shift, in the second part of the workweek. Median airborne levels during the work shift of 15 PAHs (both vapor and particulate phases), from naphthalene (NAP) to indeno(1,2,3-cd)pyrene, ranged from below 0.03 to 426 ng/m(3). Median excretion values of OH-Py in baseline, before- and end-shift samples were 228, 402, and 690 ng/L for AW and 260, 304, and 378 ng/L for CW. Lower values were found in nonsmokers compared to smokers (e.g., in AW 565 and 781 versus 252 and 506 ng/L in before-shift and end-shift samples, respectively). In all subjects a weak correlation between personal exposure to the sum of airborne 15 PAHs and OH-Py was observed (r = 0.30). The results of this article show that AW experienced a moderate occupational exposure to airborne PAHs, resulting in a significant increase of urinary OH-Py during the workday and the workweek. The contribution of working activities to internal dose was in the same order of magnitude of the contribution of cigarette smoking. PMID- 17119221 TI - Carcinogens in the construction industry. AB - The construction industry is a complex work environment. The work sites are temporary and rapidly changing. Asbestos has been widely used in construction industry, but the risks were primarily detected in specialized trades, such as insulation workers and plumbers. Today, the majority of cases related to asbestos exposure will occur in other occupational groups in the construction industry. In a large cohort of Swedish construction workers, insulators and plumbers constituted 37% of all cases of pleural mesothelioma between 1975 and 1984 while they constituted 21% of the cases between 1998 and 2002. It is estimated that 25 40% of all male cases of pleural mesothelioma in Sweden are caused by asbestos exposure in the construction trades. There are many other known carcinogens occurring in the construction industry, including PAHs, diesel exhausts, silica, asphalt fumes, solvents, etc., but it is difficult to estimate exposures and thus the size of the risk. The risk of cancer is less easy to detect with traditional epidemiological methods in the construction industry than in other industrial sectors. It is not sufficient to rely upon broad epidemiological data to estimate the risk of cancer due chemicals in the construction industry. Thus, a strategy to decrease exposure, e.g., to dust, seems a feasible way to reduce the risk. PMID- 17119222 TI - Epoxy resins in the construction industry. AB - Epoxy resins are used as coatings, adhesives, and in wood and concrete repair. However, epoxy resins can be highly irritating to the skin and are strong sensitizers. Some hardeners are carcinogenic. Based on the results of earlier Dutch studies, an international project on "best practices,"--Epoxy Code--with epoxy products was started. Partners were from Denmark, Germany, the Netherlands, and the UK. The "Code" deals with substitution, safe working procedures, safer tools, and skin protection. The feasibility of an internationally agreed "ranking system" for the health risks of epoxy products was studied. Such a ranking system should inform the user of the harmfulness of different epoxies and stimulate research on less harmful products by product developers. PMID- 17119223 TI - Health effects of hazardous waste. AB - Since 1995, the Agency for Toxic Substances and Disease Registry (ATSDR) has evaluated environmental contaminants and human health risks at nearly 3000 sites. Hazardous substances at these sites include newly emerging problems as well as historically identified threats. ATSDR classifies sites according to the degree of hazard they represent to the public. Less than 1% of the sites investigated are considered urgent public health hazards where chemical or physical hazards are at levels that could cause an immediate threat to life or health. Approximately 20% of sites have a potential for long-term human exposures above acceptable risk levels. At almost 40% of sites, hazardous substances do not represent a public health hazard. Completed exposure pathways for contaminants in air, water, and soil have been reported at approximately 30% of evaluated sites. The most common contaminants of concern at these sites include heavy metals, volatile organic compounds, and polychlorinated biphenyls. This article reviews ATSDR's ongoing work by examining the historic hazard of lead, the contemporary hazard of asbestos, and the emerging issue of perchlorate contamination. PMID- 17119224 TI - Cancer mortality in an area of Campania (Italy) characterized by multiple toxic dumping sites. AB - Several recent studies have documented that a widespread practice of dumping toxic wastes has taken place for many years in the Provinces of Naples and Caserta. Extensive programs of environmental monitoring are currently ongoing in the area. In this frame, the Department of Civil Defence of the Italian Government has appointed an ad hoc study group in order to assess the health status of the population resident in the area of interest. The first investigation performed by the study group has been a geographic study on cancer mortality and occurrence of malformations in 196 municipalities constituting the two Provinces. The study detected an area located in the southeastern part of the Province of Caserta and in the northwestern part of the Province of Naples, where cancer mortality and congenital malformations show significantly increased rates with respect to expected figures derived from the regional population. The area highlighted by the study is, in general terms, overlapping with the area where most illegal dumping of toxic wastes took place. It is now recommended that mortality studies be extended to take into account other health outcomes, to search for correlations with environmental exposures, and consider possible confounding factors. PMID- 17119225 TI - Hazardous waste: recognition of the problem and response. AB - Two accidents, Seveso 1976 and Bhopal 1984, paved the way toward the international control of hazardous substances and waste, and public dissemination of information on their releases and transfers. The final facility cleanup in Seveso took place in 1982. The remaining dioxin-containing residues were packed in 41 drums. However, the drums disappeared and were found several months later in France. This was the starting point for the development of international control on transboundary movements of waste within the OECD. Between 1983 and 1992 OECD developed five binding Council Acts on the classification and control of transboundary movements of waste. This work also provided the foundation for the adoption of a series of directives and the Shipment Regulation 259/93 within the European Community, aimed at monitoring and controlling transboundary movements of hazardous waste, and of a global instrument in 1989 under UNEP: Basel Convention on the Control of Transboundary Movements of Hazardous Waste and Their Disposal. The Bhopal 1984 and another similar accident shortly thereafter led to the development of a mandatory Toxics Release Inventory (TRI) in the United States in 1986, to inform the public of releases and transfers of potentially hazardous pollutants and waste. OECD work has supported the rapidly increasing implementation of Pollutant Release and Transfer Registers (PRTRs) since 1994. The Basel 1989, Rotterdam 1998, and Stockholm 2001 Conventions all require strict control of the production and management of hazardous chemicals and waste. Yet, production of hazardous chemicals and generation of hazardous waste are still in rise. PMID- 17119226 TI - New directions in managing hazardous wastes from an industry perspective. AB - In past decades waste was left to the lowest level of staff to dispose of at the lowest cost. Today, business managers in leading companies consider waste to be part of their responsibility. There is money to be made by eliminating waste problems. Leading companies are moving to the top of the waste disposal hierarchy and eliminating waste for themselves and their customers. PMID- 17119227 TI - Developments in management and technology of waste reduction and disposal. AB - Scandals and public dangers from the mismanagement and poor disposal of hazardous wastes during the 1960s and 1970s awakened the modern-day environmental movement. Influential publications such as "Silent Spring" and high-profile disposal failures, for example, Love Canal and Lekkerkerk, focused attention on the use of chemicals in everyday life and the potential dangers from inappropriate disposal. This attention has not abated and developments, invariably increasing expectations and tightening requirements, continue to be implemented. Waste, as a surrogate for environmental improvement, is a topic where elected representatives and administrations continually want to do more. This article will chart the recent changes in hazardous waste management emanating from the European Union legislation, now being implemented in Member States across the continent. These developments widen the range of discarded materials regarded as "hazardous," prohibit the use of specific chemicals, prohibit the use of waste management options, shift the emphasis from risk-based treatment and disposal to inclusive lists, and incorporate waste producers into more stringent regulatory regimes. The impact of the changes is also intended to provide renewed impetus for waste reduction. Under an environmental control system where only certainty is tolerated, the opportunities for innovation within the industry and the waste treatment and disposal sector will be explored. A challenging analysis will be offered on the impact of this regulation-led approach to the nature and sustainability of hazardous waste treatment and disposal in the future. PMID- 17119228 TI - Systematic approach to evaluating trade-offs among fuel options: the lessons of MTBE. AB - The fuel additive methyl tertiary butyl ether (MTBE) has been used in an effort to improve air quality in the United States, but other undesirable effects, particularly the contamination of water resources, were eventually judged to outweigh any air quality benefits it may have offered. The experience with MTBE offers many lessons, including the need to evaluate potential positive and negative environmental impacts associated with fuel choices using a comprehensive approach that combines a product life-cycle perspective with the risk assessment paradigm. Such an approach, referred to as "comprehensive environmental assessment" (CEA), is illustrated here by highlighting some of the issues that might be considered in evaluating reformulated gasoline (RFG) produced with MTBE, ethanol, or no oxygenate. PMID- 17119229 TI - Environmental impacts and costs of energy. AB - Environmental damage is one of the main justifications for continued efforts to reduce energy consumption and to shift to cleaner sources such as solar energy. In recent years there has been much progress in the analysis of environmental damages, in particular thanks to the ExternE (External Costs of Energy) Project of the European Commission. This article presents a summary of the methodology and key results for the external costs of the major energy technologies. Even though the uncertainties are large, the results provide substantial evidence that the classical air pollutants (particles, No(x), and SO(2)) from fossil fuels impose significant public health costs, comparable to the cost of global warming from CO(2) emissions. The total external costs are relatively low for natural gas (in the range of about 0.5-1 eurocents/kWh for most EU countries), but much higher for coal and lignite (in the range of about 2-6 eurocents/kWh for most EU countries). By contrast, the external costs of nuclear, wind, and photovoltaics are very low. The external costs of hydro are extremely variable from site to site, and the ones of biomass depend strongly on the specific technologies used and can be quite large for combustion. PMID- 17119230 TI - Worldwide governmental efforts to locate and destroy chemical weapons and weapons materials: minimizing risk in transport and destruction. AB - The article gives an overview on worldwide efforts to eliminate chemical weapons and facilities for their production in the context of the implementation of the 1997 Chemical Weapons Convention (CWC). It highlights the objectives of the Organisation for the Prohibition of Chemical Weapons (OPCW), the international agency set up in The Hague to implement the CWC, and provides an overview of the present status of implementation of the CWC requirements with respect to chemical weapons (CW) destruction under strict international verification. It addresses new requirements that result from an increased threat that terrorists might attempt to acquire or manufacture CW or related materials. The article provides an overview of risks associated with CW and their elimination, from storage or recovery to destruction. It differentiates between CW in stockpile and old/abandoned CW, and gives an overview on the factors and key processes that risk assessment, management, and communication need to address. This discussion is set in the overall context of the CWC that requires the completion of the destruction of all declared CW stockpiles by 2012 at the latest. PMID- 17119231 TI - Options for the destruction of chemical weapons and management of the associated risks. AB - The destruction of chemical weapons is a hazardous operation. The degree of hazard posed, however, is not uniform and is dependent on the specific chemical agent and the configuration of the weapon or bulk storage vessel in which it is contained. For example, a highly volatile nerve agent in an explosively configured munition, such as a rocket, poses a very different hazard from that of a bulk storage container of viscous mustard gas. Equally the handling of recovered, often highly corroded, World War (WW)I or WWII chemical munitions will pose a very different hazard from that associated with dealing with modern chemical weapons stored under the appropriate conditions. Over the years, a number of technologies have been developed for the destruction of chemical weapons. Each has its advantages and disadvantages. None of them provide a universal solution to the problem. When assessing options for the destruction of these weapons and the management of the associated risks, therefore, it is important to give due consideration and weight to these differences. To ensure that the destruction technology selected takes due account of them and that the resulting overall risk assessment accurately reflects the actual risks involved. PMID- 17119232 TI - Health and environmental threats associated with the destruction of chemical weapons. AB - Still existing arsenals of chemical weapons (CW) pose not only security threats for possible use in hostilities by state actors or misuse by terrorists but also safety threats to humans and biota due to leakages and possible accidents. The Chemical Weapons Convention (CWC) commits the States Parties (SPs) to destroy CW using technologies taking into consideration human health and environmental protection. It does not allow methods, routinely used up to the 1970s, such as earth burial, open-pit burning, and sea dumping. Long-term health and environmental threats and some accidents that have already occurred in the known localities of the sea-dumped and earth-buried arsenals of Nazi-German armed forces in the Baltic Region and of Imperial Japanese forces in the Far East Region are analyzed according to the impact of major CW and ammunition types (i.e., sulfur mustard--HD, tabun--GA, arsenicals--DA, DC, DM, arsine oil, and chloroacetophenone--CN). Any possible operations and handling with CW envisaged by the CWC as well as their verification are summarized taking into account the health threat they pose. CW and toxic armament waste to be destroyed and applied technologies (both developed and under current use in operational CW destruction facilities [CWDF]) are reviewed as are systems of health safety and environmental protection of the destruction/demilitarization stems from the extraordinary high toxicity of supertoxic lethal agents in man and biota. Problems of currently used Russian and U.S. standards for maximum allowable workplace concentrations and general population limits and possibilities of their determination by available analytical instrumentation are discussed. PMID- 17119233 TI - Results of long-term carcinogenicity bioassay on Sprague-Dawley rats exposed to aspartame administered in feed. AB - Aspartame (APM) is one of the most widely used artificial sweeteners in the world. Its ever-growing use in more than 6000 products, such as soft drinks, chewing gum, candy, desserts, etc., has been accompanied by rising consumer concerns regarding its safety, in particular its potential long-term carcinogenic effects. In light of the inadequacy of the carcinogenicity bioassays performed in the 1970s and 1980s, a long-term mega-experiment on APM was undertaken at the Cesare Maltoni Cancer Research Center of the European Ramazzini Foundation on groups of male and female Sprague-Dawley rats (100-150/sex/group), 8 weeks old at the start of the experiment. APM was administered in feed at concentrations of 100,000, 50,000, 10,000, 2,000, 400, 80, or 0 ppm. Treatment lasted until spontaneous death of the animals. The results of the study demonstrate that APM causes: (a) an increased incidence of malignant tumor-bearing animals, with a positive significant trend in both sexes, and in particular in females treated at 50,000 ppm (P < or = 0.01) when compared to controls; (b) an increase in lymphomas-leukemias, with a positive significant trend in both sexes, and in particular in females treated at doses of 100,000 (P < or = 0.01), 50,000 (P < or = 0.01), 10,000 (P < or = 0.05), 2000 (P < or = 0.05), and 400 ppm (P < or = 0.01); (c) a statistically significant increased incidence, with a positive significant trend, of transitional cell carcinomas of the renal pelvis and ureter in females and particularly in those treated at 100,000 ppm (P < or = 0.05); and (d) an increased incidence of malignant schwannomas of the peripheral nerves, with a positive trend in males (P < or = 0.05). The results of this mega experiment indicate that APM, in the tested experimental conditions, is a multipotential carcinogenic agent. PMID- 17119234 TI - Results of a long-term carcinogenicity bioassay on Sprague-Dawley rats exposed to sodium arsenite administered in drinking water. AB - Arsenic (As) is a metal found in nature whose acute and chronic toxic effects have been known for decades. Hundreds of millions of people are at risk of exposure to As and its various chemical forms which can occur in the occupational and general environment in air, water, soil, food, and medicines. Several epidemiological studies have shown that prolonged exposure to As can induce various types of malignant tumors in humans, namely, skin, lung, liver, kidney, and bladder cancers. These effects have been observed particularly in geographic areas where people are exposed to well water with high concentrations of As. While the risks of As at high concentrations are well documented, there is still a great deal of uncertainty regarding the risk of exposure to As at very low levels. This uncertainty is due to the absence of adequate epidemiological data and the insufficiency of experimental data currently available. Given the limited evidence demonstrating the carcinogenic potential of As in animals, a long-term carcinogenicity bioassay on sodium arsenite (NaAsO(2)) was performed at the Cesare Maltoni Cancer Research Center (CMCRC) of the European Ramazzini Foundation (ERF). NaAsO(2) was administrated with drinking water at concentrations of 200, 100, 50, or 0 mg/L, for 104 weeks to Sprague-Dawley rats (50/sex/group), 8 weeks old at the start of the study. The animals were monitored until spontaneous death at which time each animal underwent complete necropsy. Histopathological evaluation of all pathological lesions and of all organs and tissues collected was routinely performed on each animal. The results demonstrate that in our experimental conditions NaAsO(2) induces sparse benign and malignant tumors among treated rats. The types of tumors observed are infrequent in the strain of Sprague-Dawley rats of the colony used in our laboratory, namely, lung adenomas and carcinomas, kidney adenomas/papillomas and carcinomas, and bladder carcinomas. Notably, an elevated incidence of these types of oncological lesions is also observed among people living in geographical areas where As is present at higher concentrations in drinking water. PMID- 17119235 TI - Use of carcinogenicity bioassays in the IARC monographs. AB - Carcinogenicity bioassays generally provide the best means of assessing the potential for a chemical to be a human carcinogenic hazard. The results of carcinogenicity bioassays are usually the key determinants of IARC Monograph evaluations. Along with carcinogenicity bioassays and epidemiological studies, the International Agency for Research on Cancer (IARC) also encourages the consideration of mechanistic data and other relevant data. During 2005 IARC is updating the Preamble to the IARC Monographs, which describes the principles and procedures used in developing the Monographs, including the criteria that guide the evaluations. Proposed revisions to the Preamble make some changes in the criteria for evaluating carcinogenicity in experimental animals to reflect the greater confidence that can be placed in Good Laboratory Practice (GLP) studies. Other changes will give more specific guidance for the evaluation of mechanistic data. Sections on mechanistic data will be given more prominence in future Monographs and will be more closely linked with toxicokinetics. Future Monographs will also include a new section on susceptible individuals, populations, and life stages that will often be based on the understanding of mechanisms. In addition, the draft Preamble discusses IARC's procedures for promoting impartial evaluations by avoiding conflicts of interests and ensuring that working groups are free from interference. PMID- 17119236 TI - Chemical hazards in health care: high hazard, high risk, but low protection. AB - It is counter-intuitive that the healthcare industry, whose mission is the care of the sick, is itself a "high-hazard" industry for the workers it employs. Possessing every hazard class, with chemical agents in the form of pharmaceuticals, sterilants, and germicidals in frequent use, this industry sector consistently demonstrates poor injury and illness statistics, among the highest in the United States, and in the European Union (EU), 34% higher than the average work-related accident rate. In both the United States and the EU, about 10% of all workers are employed in the healthcare sector, and in developing countries as well, forecasts for the increasing need of healthcare workers (HCW) suggests a large population at potential risk of health harm. The explosion of technology growth in the healthcare sector, most obvious in pharmaceutical applications, has not been accompanied by a stepped up safety program in hospitals. Where there is hazard recognition, the remedies are often voluntary, and often poorly enforced. The wrong assumption that this industry would police itself, given its presumed knowledge base, has also been found wanting. The healthcare industry is also a significant waste generator threatening the natural environment with chemical and infectious waste and products of incineration. The ILO has recommended that occupational health goals for industrial nations focus on the hazards of new technology of which pharma and biopharma products are the leaders. This unchecked growth cannot continue without a parallel commitment to the health and safety of workers encountering these "high tech" hazards. Simple strategies to improve the present state include: (a) recognizing healthcare as a "high-hazard" employment sector; (b) fortifying voluntary safety guidelines to the level of enforceable regulation; (c) "potent" inspections; (d) treating hazardous pharmaceuticals like the chemical toxicants they are; and (e) protecting HCWs at least as well as workers in other high-hazard sectors. PMID- 17119237 TI - National and international response to occupational hazards in the healthcare sector. AB - The health care sector is one of the largest, most rapidly expanding areas of employment and is increasingly in need of qualified staff especially in the area of nursing. The health care sector is complex and comprises a variety of largely different professions; occupational hazards and exposures differ accordingly. Rates of absenteeism, reported work-related ill-health, and early retirement or departure from professions are comparatively high, especially among the nursing staff. While classical health hazards are addressed by international and national regulations, underlying causes of ill-health and departure from the profession, such as psychological stress, violence, pressing time schedules, and poor work organization are less well heeded. Practical guidance and quality information have become increasingly available from national and international Occupational Safety and Health (OSH) institutions and can be easily accessed through the Internet. They will undoubtedly benefit the profession, but difficulties not related to OSH will warrant political solutions. This presentation provides access to relevant international and European Union (EU) legislation and to valuable information resources for health care workers available from the Internet. PMID- 17119238 TI - Hazardous anticancer drugs in health care: environmental exposure assessment. AB - Exposure of healthcare workers to anticancer drugs became problematic in the 1970s. Shortly thereafter, studies began documenting exposure of healthcare workers to these drugs. Investigations employing biological markers, such as urine mutagenicity, chromosomal aberrations, sister chromatid exchanges, and micronuclei, demonstrated associations between occupational exposures and elevated marker levels. Other analytical methods emerged to monitor workplaces where drugs were handled. These contemporary studies uncovered widespread contamination of drugs on work surfaces, trace amounts in air samples, and their presence in the urine of workers. Vials containing these drugs are often contaminated with the drug when they are shipped. Most workplace surfaces are contaminated with the drugs being prepared and used in that area. Other anticancer/hazardous drugs would most likely be used in these areas. The interior surfaces of biological safety cabinets and isolators, floors, countertops, carts, storage bins, waste containers, treatment areas, tabletops, chairs, linen, and other items are all potential sources of exposure to anticancer drugs. Patient body fluids contain the drugs and/or metabolites, often more biologically active than the parent compounds. An exposure assessment of areas where anticancer/hazardous drugs are handled must consider every potential source and route of exposure. Data from surface contamination and inhalation studies suggest that dermal exposure is the primary route of exposure. Assessment of exposure is the first step in providing a safe work environment for these workers. However, because of the many drugs to which they are exposed, any assessment can only be an estimation of the overall exposure. PMID- 17119239 TI - Beyond managing healthcare risks: the health-promoting hospital initiative in Mexico. AB - The hospital industry is unique for having within it "customers" exposed to a complex mix of risks. A model is proposed that combines both the risk assessment and the promoting hospital models. This model acts in three stages: exposure elimination and protection, health aptitudes and culture, and hospital population action, and includes specific operations that can be tracked through specific effectiveness factors. Being tested in a small community hospital, there is an opportunity to apply it within the current Mexican Health reform that moves the financial risk from the patient to the provider and thus may support health promotion. PMID- 17119240 TI - Handling anticancer drugs: from hazard identification to risk management? AB - The potential health hazards involved with antineoplastic agents have been known for decades. Many anticancer drugs are recognized as carcinogens and genotoxins in experimental assays. Second cancers have been recorded in follow-up studies with treated patients. The first findings on occupational exposures among hospital personnel administering chemotherapy were reported only in 1979. Since that time a magnitude of studies have been published using various chemical and biological exposure measurements. These findings prompted improvements in the handling practices of personnel working with anticancer drugs. In spite of strict guidelines for the safe handling of cancer chemotherapeutic agents and extensive improvements in the handling facilities in hospitals, also recent studies have revealed detectable, even if generally much decreased, amounts of indicator drugs in air and surface wipe samples, also including biological samples of personnel in hospital pharmacies and cancer therapy wards. Follow-up measurements show that application of strict safety precautions in hospitals decreases the biological exposure and/or effect markers to the level of unexposed referents. Open information and constant tutoring of personnel to avoid the hazards when working with anticancer drugs is absolutely necessary with the increasing use of these important drugs. PMID- 17119241 TI - Chemical safety and health conditions among Hungarian hospital nurses. AB - In the present study genotoxicological and immunotoxicological follow-up investigations were made on 811 donors including 94 unexposed controls and 717 nurses with various working conditions from different hospitals (The Hungarian Nurse Study). The nurses were exposed to different chemicals: cytostatic drugs, anesthetic, and sterilizing gases, such as ethylene oxide (ETO) and formaldehyde. The measured biomarkers were: clinical laboratory routine tests, completed with genotoxicological (chromosome aberrations [CA], sister chromatid exchange [SCE]), and immune-toxicological monitoring (ratio of lymphocyte subpopulations, lymphocyte activation markers, and leukocyte oxidative burst). The highest rate of genotoxicologically affected donors (25.4%) was found in the group of cytostatic drug-exposed nurses. Comparing geno- and immunotoxicological effect markers, we found that among genotoxicologically affected donors the frequency of helper T cell (Th) lymphocytes, the ratio of activated T and B cells increased, whereas the oxidative burst of leukocytes decreased. In hospitals with lack of protective measures increased CA yields were observed compared to those with ISO 9001 quality control or equivalent measures. Anemia, serum glucose level, thyroid dysfunctions, benign, and malignant tumors were more frequent in the exposed groups than in controls. The hygienic standard of the working environment is the basic risk factor for the vulnerability of nurses. On the basis of these results, it is suggested, that the used cytogenetic and immunological biomarkers are appropriate to detect early susceptibility to diseases. The Hungarian Nurse Study proved that the use of safety measures could protect against occupational exposure at work sites handling cytostatic drugs, anesthetic, and sterilizing gases. PMID- 17119242 TI - Residual hazard assessment related to handling of antineoplastic drugs: safety system evolution and quality assurance of analytical measurement. AB - Despite improvement of operating procedures and publication of safety guidelines, contamination is still observed in healthcare settings where antineoplastic drugs (ADs) are handled. Even after cleaning work areas, some residual contamination may still be present. Zero percent contamination is not a realistic goal, but the scientific community should set zero contamination as its main goal. The strategies to reach this objective may be traced based on the followings: (a) a wider number of drugs should be monitored; (b) safety equipment and devices must be available to the workers; (c) the likely source of widespread contamination in workplaces is the safety cabinet; (d) direct determination of the parent drug or its metabolite in urine is the recommended approach because it provides higher sensitivity and specificity; (e) reliable analytical methods are necessary to measure the extent of contamination; and (f) analytical methods intended to be applied for routine testing must be assessed through method validation studies. These studies rely on the determination of overall method performance parameters including uncertainty measurement. Our laboratory has developed and validated a number of analytical methods for the determination of several drugs in environmental and biological samples. Surveys were carried out in several hospitals, and there has been progressive, significant decrease in the number of positive samples, mostly due to the improvement of working procedures and safety measures. PMID- 17119243 TI - Framing the future in light of the past: living in a chemical world. PMID- 17119244 TI - Children's environment and health in Latin America: the Ecuadorian case. AB - Environmental health problems of children in Latin America and Ecuador are complex due to the close relationship that exists between social and environmental factors. Extended poverty and basic problems, such as the lack of drinking water and sanitation, are common. Infectious diseases are the greatest cause of morbidity and mortality among children. Development in industry and the introduction of chemical substances in agriculture add new risks including pesticide use, heavy metal exposure, and air pollution. Major problems can be divided into (a) lack of basic infrastructure, (b) poor living conditions, (c) specific environmental problems, and (d) child labor. Reproductive health disorders are frequent in developing countries like Ecuador. Issues related to children's health should consider new approaches, creative methodologies, and the search for independent predictors to separate environmental from social problems. Only with knowledge of the specific contribution of each factor, can it be possible to develop a strategy for prevention. PMID- 17119245 TI - Environmental impacts on children's health in Southeast Asia: genotoxic compounds in urban air. AB - Air pollution is a serious problem in many countries in Southeast Asia, particularly in major metropolises with high levels of traffic congestion generating significant amounts of genotoxic substances. The contribution of such environmental exposure to children's illnesses, such as respiratory diseases and cancer, is a public health concern. Inner-city children may have higher levels of exposure to genotoxic substances in the air than those living in rural areas. This study was conducted in Bangkok, where ambient levels of polycyclic aromatic hydrocarbons (PAHs) and benzene are relatively high. Bangkok school children were exposed to total PAHs at about sixfold higher levels than those in rural areas, with levels of urinary 1-hydroxypyrene (1-OHP) also being significantly higher. PAH-DNA adduct levels in lymphocytes were fivefold higher in Bangkok children. Benzene exposure in Bangkok school children was more than twofold higher than the levels measured in children from the rural areas. This is in agreement with the biomarkers of internal dose, that is, blood benzene and urinary trans, trans muconic acid (t,t-MA) levels. The potential health risks from exposure to PAHs and benzene were assessed through the use of DNA damage and DNA repair capacity as markers of early biological effect. DNA strand breaks were significantly higher in Bangkok school children, while DNA repair capacity was significantly lower. It appears that children in major cities in developing countries may have an increased health risk for the development of certain diseases, such as cancer due to exposure to genotoxic substances in their environment. PMID- 17119246 TI - New developments in children's environmental health in Europe. AB - Important developments have taken place in Europe regarding children's environmental health (CEH) over the last few years. In 1999 the Third Ministerial Conference on Environment and Health identified CEH as a priority area and started a process of scientific review and policy development that culminated at the Fourth Ministerial Conference held in Budapest in June 2004 with the adoption of the Children's Environment and Health Action Plan for Europe (CEHAPE). The rationale of the CEHAPE is based on a thorough review of the scientific evidence on CEH and on a study that quantified for the first time the burden of disease related to the main environmental exposures of children and adolescents in Europe. The Action Plan suggests actions and policies to achieve the four main priority goals: clean air, safe water, chemical and physical agents, and injuries. Over the same period, the European Commission (EC) has strengthened its focus on environment and health issues, has supported research on CEH, and has developed a proposal for a new EU regulatory framework for chemicals that has clear implications for children and for the reproductive period. The proposed new system, called REACH (Registration, Evaluation, and Authorization of Chemicals), currently under examination by the European Parliament, aims at reducing risks to human health and improvement of environmental quality through the better and earlier identification of the properties of chemical substances. The EC also adopted policies and action plans that are very relevant to children, such as the EU European Environment and Health Strategy, referred to as the SCALE initiative (Science, Children, Awareness, Legislation, Evaluation), and the 2004-2010 Environment and Health Action Plan. PMID- 17119247 TI - Toxicogenomics--a new systems toxicology approach to understanding of gene environment interactions. AB - Toxicogenomics is a new interdisciplinary area of research being developed to monitor the expression of multiple genes, proteins, and metabolites simultaneously. It combines new technologies in genomics, proteomics, and metabolomics with traditional tools of pathology and toxicology to study biological response to drugs and other environmental xenobiotics. The biological response to environmental exposure is so complex and involves so many interactive factors that the use of a systems biology analytical approach is required. In my opinion, the development of the field of toxicogenomics will provide powerful and relatively inexpensive tools to identify biomarkers and to relate exposure and biological events during disease progression. PMID- 17119248 TI - Toxicoproteomics in liver injury and inflammation. AB - Toxicoproteomics, in applying proteomics to toxicology, seeks to identify critical proteins and pathways in biological systems responding to adverse chemical exposures and environmental stressors using global protein expression technologies. Toxicoproteomics is being exploited for the discovery of new biomarkers and toxicity signatures in target organs, such as liver, in major biological processes, such as inflammation, in mapping serum, plasma, and other biofluid proteomes, and in parallel proteomic and transcriptomic studies. The new field of toxicoproteomics is uniquely positioned toward discovery of new biomarkers and signatures of tissue injury and a better understanding of protein expression responses during toxicity and environmental disease. PMID- 17119249 TI - Gene expression alterations in immune system pathways following exposure to immunosuppressive chemicals. AB - Exposure to environmental agents can affect a number of adverse immunological outcomes, including changes in the incidence of infectious disease. Diethylstilbestrol (DES), dexamethasone (DEX), cyclophosphamide, and 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) are immunosuppressive chemicals that can induce similar pathophysiological end points in the thymus; however, the mechanism of toxicity is different for each compound. We examined differential gene expression in the spleen and thymus following chemical exposure and correlated these changes with alterations in functional immune end points and our knowledge of the known mechanisms of action. RNA from the spleen and thymus has been analyzed using Illumina Sentrix arrays and BeadStudio software. Preliminary data suggest that DES induced the greatest number of gene changes in the spleen, while DEX induced the most changes in the thymus. In both spleen and thymus, genomic analysis revealed gene expression changes that were common to multiple chemicals and that may be associated with xenobiotic-induced immune system perturbations, including alterations in genes associated with apoptosis, antigen processing and presentation, and response to biotic stimulus. This was particularly evident in the thymus, where there were many similarities in the expression profiles, as well as gene alterations unique to a single compound. In contrast, expression profiles in spleen were more distinct. The category of genes most profoundly affected by all four chemicals was response to biotic stimulus: there were both clusters of genes modulated by multiple chemicals and genes altered by a single chemical. The distinct gene profiles may specifically relate to cellular targets and mechanism of action. PMID- 17119250 TI - Transcriptional profiling and functional genomics reveal a role for AHR transcription factor in nephrogenesis. AB - Transcriptional profiling and functional genomics experiments using E11.5 metanephros organ cultures from Ahr(-/-) and Ahr(+/+) have shown that aryl hydrocarbon receptor (AHR) transcription factor is involved in the regulation of mesenchymal-to-epithelial transition (MET) during nephrogenesis. This response is mediated by alterations in the post-transcriptional control of Wilms' tumor suppressor (Wt1) gene and Wt1 splicing. In this article, biologically relevant gene predictor sets of the nephrogenic response were calculated for target genes of interest. The predictability of the gene set for each target was quantified by the coefficient of determination which provided a good criterion for identification of predictor sets that define the complex gene-gene interactions co-regulated by Ahr and Wt1. A subset of the signature genes was found to be co regulated by Ahr and Wt1 and was responsible for shifts in renal cell transdifferentiation. PMID- 17119251 TI - Results of long-term carcinogenicity bioassays on Coca-Cola administered to Sprague-Dawley rats. AB - Coca-Cola was invented in May 1886 in Atlanta, Georgia by a pharmacist who, by accident or design, mixed carbonated water with the syrup of sugar, phosphoric acid, caffeine, and other natural flavors to create what is known as "the world's favorite soft drink." Coca-Cola is currently sold in more than 200 countries and in early 2000, the company sold its 10 billionth unit case of Coca-Cola branded products. Given the worldwide consumption of Coca-Cola, a project of experimental bioassays to study its long-term effects when administered as substitute for drinking water on male and female Sprague-Dawley rats was planned and executed. The objective of the project was to study whether and how long-term consumption of Coca-Cola affects the basic tumorigram of test animals. The bioassays were performed on rats beginning at different ages, namely: (a) on males and females exposed since embryonic life or from 7 weeks of age; and (b) on males and females exposed from 30, 39, or 55 weeks of age. Overall, the project included 1999 rats. During the biophase, data were collected on fluid and feed consumption, body weight, and survival. Animals were kept under observation until spontaneous death and underwent complete necropsy. The results indicate: (a) an increase in body weight in all treated animals; (b) a statistically significant increase of the incidence in females, both breeders and offspring, bearing malignant mammary tumors; (c) a statistically significant increase in the incidence of exocrine ademonas of the pancreas in both male and female breeders and offspring; and (d) an increased incidence, albeit not statistically significant, of pancreatic islet cell carcinomas in females, a malignant tumor which occurs very rarely in our historical controls. On the basis of the results of this study, excessive consumption of regular soft-drinks should be generally discouraged, in particular for children and adolescents. PMID- 17119252 TI - Occupational kidney cancer: exposure to industrial solvents. AB - We report seven cases of renal cell carcinoma in workers diagnosed with occupational exposure via skin contact and inhalation to industrial solvents containing benzene. The clinical significance of these cases are: (a) all seven patients diagnosed with kidney cancer were seen by private physicians who missed addressing occupational history to industrial solvents; (b) emphasize the importance of taking an in-depth history including occupational history in any patient presented to the clinician, especially like in these cases, kidney cancer; and (c) demonstrate the importance of educating workers. We believe that there exist more patients with renal cancer whose diagnosis has been rendered "idiopathic" due to the lack of detailed occupational, environmental, personal, and family history. PMID- 17119253 TI - Occupation and breast cancer: a Canadian case-control study. AB - A local collaborative process was launched in Windsor, Ontario, Canada to explore the role of occupation as a risk factor for cancer. An initial hypothesis generating study found an increased risk for breast cancer among women aged 55 years or younger who had ever worked in farming. On the basis of this result, a 2 year case-control study was undertaken to evaluate the lifetime occupational histories of women with breast cancer. The results indicate that women with breast cancer were nearly three times more likely to have worked in agriculture when compared to the controls (OR = 2.80 [95% CI, 1.6-4.8]). The risk for those who worked in agriculture and subsequently worked in automotive-related manufacturing was further elevated (OR = 4.0 [95% CI, 1.7-9.9]). The risk for those employed in agriculture and subsequently employed in health care was also elevated (OR = 2.3 [95% CI, 1.1-4.6]). Farming tended to be among the earlier jobs worked, often during adolescence. While this article has limitations including the small sample size and the lack of information regarding specific exposures, it does provide evidence of a possible association between farming and breast cancer. The findings indicate the need for further study to determine which aspects of farming may be of biological importance and to better understand the significance of timing of exposure in terms of cancer risk. PMID- 17119254 TI - Adverse health effects of fluoro-edenitic fibers: epidemiological evidence and public health priorities. AB - Subsequent to the detection of a cluster of mesothelioma cases in the Sicilian town of Biancavilla, located at the slopes of Etna volcano, ad hoc epidemiological studies and environmental monitoring suggested an etiological role of an asbestiform fiber present in a stone quarry. The fiber was shown to constitute a new mineral species named fluoro-edenite. Fluoro-edenitic fibers were found in the materials extracted from the quarry and used in the local building industry, as well as in soils. Besides the risk of mesothelioma, residents in Biancavilla showed a significantly increased mortality from chronic obstructive pulmonary disease, which was particularly evident among women. In the light of these findings, Biancavilla was defined a site of national interest for environmental reclamation. The first preventive action involved termination of quarrying activity, covering with asphalt of roads previously paved with local soil materials, and removal of sources of dust in the urban area. Concurrent to the implementation of environmental cleanup, some specific "second generation" studies are now being designed and performed, namely morbidity surveys based on hospital discharge cards, monitoring of fibers in sputum and health surveillance in selected population groups. In this frame, special emphasis is given to the issue of communication, both to the general public and to target groups like family doctors, teachers, and media professionals. This experience could represent a useful basis for the elaboration of a strategy to approach similar environmental issues. PMID- 17119255 TI - Glycol ethers: a ubiquitous family of toxic chemicals: a plea for REACH regulation. AB - Glycol ethers (GE) are chemicals used since the 1930s as solvents in paints, inks, varnishes, and cleaning agents, mainly in water-based products, cosmetics, and drugs. World production approximates 1 million tons. Nineteen GE are produced or imported each year; over 1000 tons in European Union (EU) have been classified as high production volume chemicals (HPVCs). First animal data were published in 1971 and 1979 showing severe reprotoxicity for some GE. Two alerts were launched in the United States in 1982 and 1983, but the first partial GE regulation only occurred in 1993 in the EU. Although these chemicals may expose a very large population, basic toxicity data, more especially carcinogenicity, are still lacking (3/32 GE). However, experimental data were sufficient to lead developmental toxicity risk assessment since the early 1980s. Risk indices over 1000 have been calculated for consumers and workers exposed to reprotoxic GE in domestic and industrial activities. The first ban was decided in 1999 in France, but was only for drugs and cosmetics. Not surprisingly, since the late 1980s, human studies have found results similar to those in animal data: spontaneous abortions, malformations, testicular toxicity, and hematotoxicity. Despite this highly coherent set of data, and although substitution products are available, reprotoxic GE have been and still remain widely used in the world. The case of GE shows the failure of the present system based on a posteriori risk assessment. This pleads for the change of paradigm through the European REACH regulation based on the "No data, no market" principle. Ethics in REACH management should also be considered. PMID- 17119256 TI - Controlling exposure to chemicals: a simple guide. AB - Controlling exposure to chemicals in the workplace has been made easier by the use of a guide published by the U.K. Health and Safety Executive (HSE). Known as COSHH (Control of Substances Hazardous to Health Regulations) Essentials, the guide is a simple five-step procedure to devise appropriate control strategies to reduce exposures to various substances under different conditions. U.K. health and safety law requires risk assessments prior to use of hazardous substances and installation of appropriate control strategies before work commences. A 1996 survey of 1500 safety managers and trade union safety representatives revealed that the majority had little understanding of occupational safety limits for chemicals. Small- and medium-sized companies had little understanding of limits, and most could not develop control strategies. A new approach was required. COSHH Essentials is it. Developed over 3 years by a working group of hygienists and toxicologists representing HSE, industry, trade unions, and independent experts, the guide is now available in both paper-based and internet versions. It applies a hazard banding approach validated by data for 111 substances that have well founded U.K. occupational exposure limits. New users select an appropriate hazard band for chemicals based on risk phrases. Details about dustiness for powders or volatility for liquids are inserted, and the guide allocates substances to one of four exposure bands linked, in turn, to specific control strategies. Now accessible through the HSE web site, COSHH Essentials will offer control strategies for both single chemicals and whole processes. To date over 300,000 risk assessments have been carried out using the internet version of COSHH Essentials. PMID- 17119257 TI - Progress of epidemiological and molecular epidemiological studies on benzene in China. AB - Benzene is an organic solvent that has been used in industry for about 100 years throughout the world. Since 1973, a series of toxicological and molecular epidemiological studies on benzene were conducted by researchers at the Chinese Academy of Preventive Medicine (CAPM) (1973-1986) and subsequently by a collaboration between the CAPM and the National Cancer Institute (NCI) in the United States that began in 1986, which was joined by investigators from the University of California at Berkeley, the University of North Carolina at Chapel Hill, and New York University. The findings demonstrated that the risk of leukemia and lymphoma among benzene-exposed workers was significantly increased, with elevated risks for leukemia present not only at higher exposure but also among workers exposed to under 10 ppm. Therefore, the benzene permissible level was decreased to 1.8 ppm (6 mg/m(3)) and benzene-induced leukemia is treated as an occupational cancer in China. The benzene permissible level is 1.0 in the United States and in several other developed countries and it has been suggested to be decreased to 0.5 ppm (ACGIH). A number of potential biomarkers are related to benzene exposure and poisoning. Some of these are benzene oxide-protein adducts, chromosome aberration of lymphocytes, and GPA mutations in erythrocytes, a decrease in B cell and CD4(-)T cell counts in peripheral blood, and altered expression of CXCL16, ZNF331, JUN, and PF4 in lymphocytes. Variation in multiple benzene metabolizing genes may be associated with risk of benzene hematotoxicity, including CYP2E1, MPO, NQO1, and GSTT1. PMID- 17119258 TI - Carcinogen exposure and epigenetic silencing in bladder cancer. AB - Tobacco smoking, certain occupational exposures, and exposure to inorganic arsenic in drinking water have been associated with the occurrence of bladder cancer. However, in these tumors the exposure-associated pattern of somatic alterations in genes in the causal pathway for disease has been poorly characterized. Animal and in vitro studies have suggested that arsenic, tobacco carcinogens, and other exposures may act through epigenetic mechanisms. We, therefore, examined, in a population-based study of human bladder cancer (n = 351), the relationship between epigenetic silencing of the tumor-suppressor genes, p16(INK4A), RASSF1A, PRSS3, and the four SFRP genes and exposure to both tobacco and arsenic in bladder cancer. Promotor methylation silencing of each of these genes occurred in approximately 30-50% of bladder cancers. Epigenetic silencing of RASSF1A and PRSS3 and any of the SFRP genes were each significantly associated with advanced tumor stage (P < 0.001, P < 0.04, and P < 0.005, respectively). Arsenic exposure, measured as toenail arsenic, was associated with RASSF1A (P < 0.02) and PRSS3 (P < 0.1) but not p16(INK4A) or SFRP promotor methylation, in models adjusted for stage and other risk factors. Cigarette smoking was associated with a greater than twofold increased risk of promotor methylation of the p16(INK4A) gene, with greater risk seen in patients with exposures more recent to disease diagnosis, and smoking was also significantly associated with any SFRP gene methylation (P < 0.01). These results from human bladder tumors, add to the body of animal and in vitro evidence that suggests bladder carcinogens play a crucial role in the induction of important epigenetic alterations. PMID- 17119259 TI - Causal relationship from exposure to chemicals in oil refining and chemical industries and malignant melanoma. AB - Malignant melanoma has been thought to be related mainly to exposure to the sun or radiation. A review of the scientific literature reveals many significant correlations between benzene and benzene-containing solvents in the workplace and the occurrence of malignant melanoma, particularly in sites that have never been exposed to sunlight. A comparison of positive correlations between such exposure and malignant melanoma by independent investigators and negative findings by investigators with industry affiliations reveals that this difference, at least in part, may account for the discrepant findings. Based on independent studies, it is reasonable to conclude that malignant melanoma is causally related to employment-related chemical exposures in the petroleum refining industry. PMID- 17119260 TI - A regional approach to assess the impact of living in a chemical world. AB - In the United States, some 80,000 commercial and industrial chemicals are now in use of which over 30,000 are produced or used in the Great Lakes region. Thus, the environmental quality within the Great Lakes basin has been compromised particularly with respect to persistent toxic substances (PTS). Information derived from wildlife studies, prospective epidemiological and toxicological studies, databases, demographics, and Geographical Information Systems (GIS) demonstrate significant public health implications. Studies of human populations indicate: (a) elevated body burden levels of PTSs, (b) decrease in gestational age, (c) low birth weight (LBW), (d) greater risk of male children with birth defects (OR = 3.01), (e) developmental and neurological deficits, (f) increased risk of infertility, (g) changes in sex ratio, and (h) fluctuations in thyroid hormones. These findings have been identified in vulnerable populations, such as the developing fetus, children, minorities, and men and women of reproductive age who are more susceptible because of their physiologic sensitivity and/or elevated exposure to toxic chemicals. Typically such health effects are assessed on a chemical specific basis; however, most human populations are exposed to hazardous chemicals as mixtures in air, water, soil, and biota. In this article we present an assessment of the potential for joint toxic action of these substances in combinations in which they are typically found. These evaluations represent an integration of all available scientific evidence in accordance with the "NAS paradigm" for risk assessment. In aggregate, our evaluations have demonstrated a need for community-based frameworks and computational techniques to track patterns of environmentally related exposures and associated health effects. PMID- 17119261 TI - Low-dose risk, hormesis, analogical and logical thinking. AB - The hormesis theory proposes the low-dose beneficial and high-dose detrimental pattern, existing for specific conditions, as a "general default assumption" for toxicology and carcinogenicity. Crump and Kitchin and Drane underline that in a post hoc retrospective scientific literature searching for hormetic dose-response patterns, the consideration of the whole available relevant studies is necessary and, for statistical testing purposes, for instance at a 0.05 standard level, a P value obtained from 1 - (1 - P)(n) = 0.05 (i.e., P = 0.0005 for 100 examined cases) should be used (otherwise, by definition, 5 "positive" results are expected by chance over 100 cases). The hypothesis, based on some experimental data on rodents, by Calabrese and Baldwin, of an hormetic effect of 2,3,7,8-TCDD at the 1-10 ng/kgbw/day dose, of Na-saccharine in the < or = 1% of diet exposure range, of Cadmium Chloride in the 0-5 micromol/kg dose range, single injection, and of neutrons in the 0- to 2-rad dose range, are not confirmed, and, rather, are contradicted, when the whole relevant data presented by international and national agencies are considered. As far as the radiation risk is in particular concerned, a recently published epidemiological study on more than 400,000 nuclear plant workers, co-ordinated by the IARC has indicated a small, but significant risk, at the current exposure limits, and possibly below them. Therefore, the hormesis theory-based criticism of current radiation protection criteria, assumed to be excessively conservative, is not justified. Also not justified is the assumption that "by dismissing hormesis, regulatory agencies such as U.S. EPA deny the public the opportunity for optimal health and avoidance of diseases;" rather, the contrary is here considered true. Analogical considerations are not necessarily logical ones and the single result should be considered in its whole context. PMID- 17119262 TI - Occupational injury and illness meet the labor market: lessons from labor economics about lost earnings. AB - Recent labor economics studies in the United States and Canada have demonstrated that occupational injuries and illnesses often lead to substantial lost earnings for workers and their families. Other studies have shown substantial long-term lost earnings attributable to large-scale layoffs, where no health impairment has taken place. This article uses evidence from these and other studies of apparently different situations to draw inferences about how managers' actions and public policy choices can affect the costs of occupational injuries and illnesses. Although primary prevention remains the policy of choice, reduction in the impact of workplace injuries and illnesses can decrease the costs of these events and can provide substantial benefits. This article proposes two hypotheses and discusses the evidence for each: (a) Loss of the job held at the onset of illness or injury increases time off work and exacerbates workers' lost earnings. (b) Workers' losses may be substantially reduced by policies that encourage employers to rehire people recovering from or disabled by workplace injuries and illnesses. PMID- 17119263 TI - The economic costs of health service treatments for asbestos-related mesothelioma deaths. AB - This article explores the complex and neglected picture of occupational and environmental disease healthcare costs specifically relating to asbestos. Diagnosed mesothelioma cases in Scotland in one calendar year were used to investigate the subject in greater depth. Data from UK sources on asbestos disease types recorded in 2000 and their disease treatment costs were obtained. Acute care economic costs of these diseases are estimated. One hundred and twenty diagnosed, recorded, and treated cases of asbestos-related diseases occurred in 2000 in Scotland. Mesothelioma accounted for 100 cases and directly cost Scottish National Health Service hospitals an estimated 942,038 pounds. The estimated UK figure in 2000 was at least 16,014,646 pounds because official figures for diagnosed and recorded deaths from mesothelioma are running at over 1700 a year with rises predicted for 2010 of 2000 deaths. By 2003, 50,000 people in the UK had died from diagnosed and recorded mesothelioma since records began. Earlier disease treatment costs would have been significantly lower than those in 2000 but, at 2000 prices, cost to the UK was roughly 471,019,000 pounds in acute hospital expenditure. Figures for primary care costs, including caregiver costs, are incomplete or unknown. These disease costs are substantial and have some international generalizability. Treatment patterns and costs vary greatly. Many lung cancer cases due to asbestos exposure occur globally for each mesothelioma case. Hence figures provided in this article are certain to be gross underestimates of the total health service and personal economic costs of asbestos illness and treatment in Scotland. PMID- 17119264 TI - Valuing the adult health effects of air pollution in Chinese cities. AB - China's ongoing economic growth is accompanied by a large amount of air pollution that exacts significant health and economic costs on its people. Following up on some earlier work focusing upon general mortality and child-specific health effects, this article uses a larger data set, covering more than 90 Chinese cities, along with a set of China-based epidemiological functions, to estimate some of the adult health benefits of reducing urban air pollution. Projecting future air pollution based upon current conditions, it calculates the averted mortality and morbidity effects that would result from the cleanup of particulates, sulfur dioxide, and nitrogen dioxide. The inclusion of nitrogen dioxide in our analysis is particularly important because it is a growing problem and has not been included in most of the more widely known studies that examine Chinese air pollution. Finally, the economic valuation of these pollution-related health effects is developed, using a number of recent, China-based valuation studies. PMID- 17119265 TI - Chinese workers and labor conditions from state industry to globalized factories: how to stop the race to the bottom. AB - This article discusses administrative obstacles in China that hinder the full integration of the rural population into the mainstream of development during a period of rapid industrialization. The Chinese household registration only for urban residents with its golden contents of cradle-to-grave security has become a formidable stumbling block that perpetuates the status of rural migrants as second-class citizens in their own country. Rural migrant workers are excluded from certain types of jobs and are not eligible for many benefits that urbanites have, such as health, education, and unemployment protection. These workers must also pay a number of fees and work for lower minimum wages than the local residents. With a precarious legal existence in urban areas, they are easy prey to unscrupulous officials and employers. Because they are not allowed to form independent trade unions, their best option is to vote with their feet and leave the firms with the worst conditions; this is exactly what they did from 2004. Given this situation, the debate on Corporate Social Responsibility (CSR) took a new turn with not only nongovernmental organizations (NGOs) pushing it but with a wider range of employers and, of late, Chinese officials promoting their version of CSR. In the campaign to promote minimum labor standards, the norms set down in the Social Accountability 8000 were included in the CSR, recognizing the right to free collective bargaining and free trade unions but were excluded in the Chinese version even though the World Trade Organization (WTO) agreements recognized these rights. PMID- 17119266 TI - Applying cost analyses to drive policy that protects children: mercury as a case study. AB - Exposure in prenatal life to methylmercury (MeHg) has become the topic of intense debate in the United States after the Environmental Protection Agency (EPA) announced a proposal in 2004 to reverse strict controls on emissions of mercury from coal-fired power plants that had been in effect for the preceding 15 years. This proposal failed to incorporate any consideration of the health impacts on children that would result from increased mercury emissions. We assessed the impact on children's health of industrial mercury emissions and found that between 316,588 and 637,233 babies are born with mercury-related losses of cognitive function ranging from 0.2 to 5.13 points. We calculated that decreased economic productivity resulting from diminished intelligence over a lifetime results in an aggregate economic cost in each annual birth cohort of $8.7 billion annually (range: $0.7-$13.9 billion, 2000 dollars). $1.3 billion (range: $51 million-$2.0 billion) of this cost is attributable to mercury emitted from American coal-fired power plants. Downward shifts in intellectual quotient (IQ) are also associated with 1566 (range: 115-2675) excess cases of mental retardation (MR defined as IQ < 70) annually. This number accounts for 3.2% (range: 0.2-5.4%) of MR cases in the United States. If the lifetime excess cost of a case of MR (excluding individual productivity losses) is $1,248,648 in 2000 dollars, then the cost of these excess cases of MR is $2.0 billion annually (range: $143 million-$3.3 billion). Preliminary data suggest that more stringent mercury policy options would prevent thousands of cases of MR and billions of dollars over the next 25 years. PMID- 17119268 TI - New approach in the treatment of T2DM and metabolic syndrome (focus on a novel insulin sensitizer). AB - Peroxisome Proliferator-Activate Receptors (PPARs) are transcription factors belonging to the nuclear receptor superfamily. The three PPARs (alpha, beta/delta, and gamma) are distributed differently in the different organs. PPARalpha is most common in the liver, but also found in kidney, gut, skeletal muscle and adipose tissue, while PPARbeta/delta, is fairly ubiquitous; it may be found in body tissues and brain (for myelination process and lipid metabolism in the brain). PPARgamma has 3 isoforms, such as PPARgamma 1, PPARgamma 2, and PPARgamma 3. The syndrome-X was firstly coined by Reaven in 1988 and then to be provided in 1999 by the name : the metabolic syndrome-X. This metabolic syndrome represents a "Cluster" of metabolic disorders and cardiovascular risk factors which has been collected and summarized by the author and such a cluster includes: insulin resistance/hyperinsulinemia, central obesity, glucose intolerance/DM, atherogenic dyslipidemia (increase TG, decrease HDL-cholesterol, increase Apo-B, increase small dense LDL), hypertension, prothrombotic state (increase PAI-1, increase F-VII, increase fibrinogen, increase vWF, increase adhesion molecules), endothelial dysfunction, hyperuricemia, and increased hsC-RP and cytokines. The metabolic syndrome-X may lead to the development of T2DM and coronary heart disease (CHD); insulin resistance plays pivotal roles in the progression of such a syndrome and cardiovascular diseases. Improvement of Insulin Resistance, therefore, is most likely to reduce the high cardiovascular event rate in T2DM. It has been generally accepted that Insulin Resistance (detected by HOMA-R) and Acute Insulin Response = AIR (by HOMA-B) are both usually present in T2DM. The Thiazolidinedions (TZDs) are Insulin Sensitizers (e.g Rosiglitazone = ROS, Pioglitazone = PIO) introduced into clinical practice in 1997; clinical evidence data showed that TZDs improved both HOMA-R, and HOMA B. PPARgamma can be activated by TZDs and it appears to be fundamental to the pathophysiology of diabetes mellitus i.e increase GLUT-4, increase glucokinase, decrease PEPCK, increase GLUT-4, and decreases production by fat cell of several mediators that may cause insulin resistance, such as TNFalpha and resistin. PPARgamma also mediates increased production of Adiponectin and the insulin signaling intermediate PI3K, and both actions lead to increase insulin sensitivity. A "dual PPARgamma-PPARalpha agonists" (e.g PIO, but ROS poorly activate PPARalpha) might lower glucose and modulate lipids. Thus, PIO, as a stronger "dual PPARgamma-PPARalpha agonists", shows an important therapeutic pathway in diabetes mellitus and cardiovascular diseases, even in metabolic syndrome. Current evidence suggests a close relationship between activation of PPARgamma and restoration of insulin sensitivity by reductions in TNFalpha and FFAs, and the enhancement of insulin stimulation of PI3-K Pathway and also increase adiponectin & decrease resistin. PMID- 17119269 TI - Schisandrin B-induced increase in cellular glutathione level and protection against oxidant injury are mediated by the enhancement of glutathione synthesis and regeneration in AML12 and H9c2 cells. AB - To define the relative role of reduced glutathione (GSH) synthesis and regeneration in schisandrin B (Sch B)-induced increase in cellular GSH level and the associated cytoprotection against oxidative challenge, the effects of L buthionine-[S,R]-sulfoximine (BSO, a specific inhibitor of gamma-glutamate cysteine ligase (GCL)) and 1,3-bis(2-chloroethyl)-1-nitrourea (BCNU, a specific inhibitor of glutathione reductase (GR)) treatments or their combined treatment were examined in control and Sch B-treated AML12 and H9c2 cells, without and/or with menadione intoxication. Both BSO and BCNU treatments reduced cellular GSH level in AML12 and H9c2 cells, with the effect of BSO being more prominent. The GSH-enhancing effect of Sch B was also suppressed by BSO and BCNU treatments, with the effect of the combined treatment with BSO and BCNU being semi-additive. While Sch B treatment increased the GR but not GCL activity in AML12 and H9c2 cells, it increased the cellular cysteine level. BSO treatment also suppressed the Sch B-induced increase in GR activity. BSO or BCNU treatment per se did not cause any detectable cytotoxic effect, as assessed by lactate dehydrogenase leakage, but the combined treatment with BSO and BCNU was cytotoxic, particularly in H9c2 cells. The cytotoxic effect of BSO and BCNU became more apparent following the menadione challenge. The cytoprotection afforded by Sch B pretreatment was partly suppressed by BSO or BCNU treatment, or completely abrogated by the combined treatment with BSO and BCNU. In conclusion, the results indicate that the cytoprotective action of Sch B is causally related to the increase in cellular GSH level, which is likely mediated by the enhancement of GSH synthesis and regeneration. PMID- 17119270 TI - Antioxidant and antigenotoxic activities of Angelica keiskei, Oenanthe javanica and Brassica oleracea in the Salmonella mutagenicity assay and in HCT116 human colon cancer cells. AB - Epidemiological studies indicate that consumption of green-yellow vegetables rich in chlorophyll, vitamin C, vitamin E, and carotenoids reduce the risk of cancer. We sought to examine the antigenotoxic and antioxidant properties of chlorophyll rich methanol extracts of Angelica keiskei, Oenanthe javanica, and Brassica oleracea (kale). In the Salmonella mutagenicity assay, A. keiskei caused dose dependent inhibition against three heterocyclic amine mutagens in the presence of S9, O. javanica was antimutagenic only at the highest concentration in the assay (2 mg/plate), and B. oleracea showed no consistent inhibitory activity at non toxic levels. None of the extracts were effective against three direct-acting mutagens in the absence of S9. Extracts of A. keiskei and, to a lesser extent O. javanica, inhibited two of the major enzymes that play a role in the metabolic activation of heterocyclic amines, based on ethoxyresorufin-O-deethylase and methoxyresorufin-O-demethylase assays in vitro. All three plant extracts were highly effective in assays which measured ferric reducing/antioxidant power, oxygen radical absorbance capacity, and Fe2+/H2O2-mediated DNA nicking. Finally, using the 'comet' assay, all three plant extracts protected against H2O2-induced genotoxic damage in human HCT116 colon cancer cells. These findings provide support for the antigenotoxic and antioxidant properties of chlorophyll-rich extracts of A. keiskei, O. javanica, and B. oleracea, through mechanisms that include inhibition of carcinogen activation and scavenging of reactive oxygen species. PMID- 17119271 TI - The isoflavonoid aglycone-rich fractions of Chungkookjang, fermented unsalted soybeans, enhance insulin signaling and peroxisome proliferator-activated receptor-gamma activity in vitro. AB - We investigated anti-diabetic candidates and their mechanisms from the fractions of Chungkookjang (CKJ), a traditional fermented unsalted soybean, by investigating insulin signaling, peroxisome proliferator-activated receptor (PPAR)-gamma activity and glucose-stimulated insulin secretion, in vitro. Cooked soybeans (CSB) and CKJ, fermented predominantly with Bacillus subtilis, were extracted by 70% EtOH followed by an XAD-4 column chromatography with a serial mixture of solvents comprised of MeOH and water. During fermentation, the contents of isoflavonoid aglycones were elevated, and the fractions enriched with aglycones enhanced insulin-stimulated glucose uptake in 3T3-L1 adipocytes. This increase in glucose uptake resulted from stimulating a translocation of the glucose transporter (GLUT)-4 into the plasma membrane through the phosphorylation of insulin receptor substrate (IRS)-1 and Akt. Especially, daidzein enriched fractions elevated insulin-stimulated glucose uptake by acting as PPAR-gamma agonist up to levels exhibited when 10 nM insulin is administered. Fractions containing small peptides with low polarity in CKJ slightly increased glucose stimulated insulin secretion. The data suggest that an increase in isoflavonoid aglycones in CKJ, in comparison to CSB, enhances glucose utilization via activating insulin signaling and stimulates PPAR-gamma activity in adipocytes. In addition, CKJ contains small peptides improving glucose-stimulated insulin secretion in insulinoma cells. Overall, CKJ is superior to CSB in anti-diabetic action. PMID- 17119272 TI - Regulation of Mg2+-independent Ca2+-ATPase by a low molecular mass protein purified from bovine brain. AB - The goat sperm microsomal membranes have been found to contain an Mg2+ independent Ca2+-ATPase, a low affinity but highly active enzyme sharing similarities with the SERCA family of ATPases. The present study reports the identification and characterization of a 14 kilodalton cytosolic protein from bovine brain which can act as an endogenous stimulator of the enzyme with an S50 (concentration producing 50% stimulation) of 0.8 mu molar. Kinetic analysis suggests that the stimulation is noncompetitive with respect to the substrate, and the binding site(s) of the stimulator and substrate are distinct. Binding of the stimulator to the enzyme is reversible. The stimulator increases the affinity of the enzyme for calcium as evident from a decrease in K0.5 of the enzyme for calcium in presence of the stimulator. Radioactive labeling of the enzyme with [gamma-32P]-ATP suggests that the stimulator enhances the rate of dephosphorylation of the phosphoenzyme intermediate without altering the phosphorylation reaction step. The stimulatory effect of the protein has been observed only for the Mg2+-independent form of the enzyme, the Mg2+-dependent form being unaffected. PMID- 17119273 TI - Anti-adipogenic activity of rutin in 3T3-L1 cells and mice fed with high-fat diet. AB - Polyphenolic compounds were examined for their effects on suppressing adipocyte differentiation in 3T3-L1 cells. Most polyphenolic compounds inhibited adipocyte development from 3T3-L1 cells to some extent. Among them, rutin was the most effective in suppressing adipocyte differentiation in a dosage dependant manner. Activity of glycerol-3-phosphate dehydrogenase (GPDH), which has a central position in lipogenesis in adipose cells, was also decreased by rutin addition at the induction stage. RT-PCR results demonstrated that mRNA expression of adipogenic transcription factors such as peroxisome proliferator-activated receptor-gamma (PPARgamma) and CCAAT/enhancer binding protein-alpha (C/EBPalpha) in 3T3-L1 cells were remarkably down regulated by rutin treatment. For further investigation on anti-adipogenic activities of rutin, it was orally administered (25 and 50 mg/kg b.w/daily) with high-fat diet (64.4% of total calories as fat) to C57BL/6 mice. Body weight gains were less in high-fat diet + rutin fed groups (HFR) than high-fat diet alone fed group (HF) after 4 weeks. Total cholesterol contents in blood were significantly lower in HFR groups. When mRNA expressions of PPARgamma and C/EBPalpha in hepatocytes were compared between the control HF and HFR groups, their expressions in hepatocytes of HFR groups were significantly suppressed. These results indicate that rutin inhibits adipogenic development in pre-adipocytes and hepatocytes by down regulating expressions of key adipogenic transcription factors. PMID- 17119274 TI - Activation of redox-systems of monocytes by hydrogen peroxide. AB - In this work the influence of H2O2 on the ability of human blood monocytes to generate ROS upon stimulation of cells by adhesion to glass surface and fMLP was studied using the luminol-dependent chemiluminescence (LDCL) method. Pretreatment of cells with H2O2 increased the adhesiveness of monocytes and ROS generation. Superoxide generation by cells in response to fMLP depended on the duration of pretreatment and the concentration of H2O2. The stimulatory effect on fMLP induced LDCL of cells further depended on the Ca2+ concentration in the medium and on the activities of phospholipase A2, cyclooxygenases, and Mek1/2. PMID- 17119275 TI - Induced dielectric-force-effect by 50 Hz strong electric field on living tissue. AB - The main objective of this paper is to study on the fields, resultant forces, and the dielectric substances in biologic tissues. Additionally, to unveil the possible mechanism which affects on major biological activities were examined. In the scientific literature, there are many studies conducted with low frequency Electromagnetic Fields (MFs), Static Magnetic Fields (SMFs), and Pulsed Electromagnetic Fields (PEMFs). However, we face in some cases; merely electric fields may be effective on biological functions (around 10 kV/m). Parallel plate exposure system was chosen as a model in this paper. In order to overcome the difficulty of generating virtual homogenous low frequency ac (power frequencies) or dc fields, a pair of appropriately designed parallel plates exposure model may be mighty tool. In this study, an important assumption, with deriving an equation that possible explanation for heat rise in a living tissue, has been made. Consequently it has been assumed that the field E(loc) may be responsible both for the induced and oriented polarization. PMID- 17119276 TI - Characterization of isometric contractions of rat skeletal muscle in vivo: duty cycle effects. AB - Many work related injuries stem from the exertion of skeletal muscle forces over an extended period of time. Musculoskeletal injury can be caused by muscle's inability to maintain force during occupational exposure. The goal of the present study is to test how various rest times (duty cycles) between long isometric contractions will affect decrements in force, and develop a model that characterizes force decrements due to skeletal muscle fatigue. All tests were performed in vivo on the tibialis anterior muscle of anesthetized Sprague-Dawley rats. Animals were randomly assigned to either a 10 second (N=8), 1 minute (N=8), or 5 minute (N=8) duty cycle group. All animals were then subjected to 7 isometric contractions (duration of 2.8 seconds). A model was constructed to characterize forces changes over the duration of a contraction and over multiple contractions. The model consisted of a power law and an exponential component; these two components were combined by using an exponential weighting function. Overall, the combination of a power law and exponential model with a weighting function satisfactorily characterized the changes in isometric force for the 10 second duty cycle, but a simpler exponential model could be used where longer duty cycles are performed. PMID- 17119277 TI - Thermal processing and characterization of 316LVM cardiovascular stent. AB - In the current investigation, annealing was employed as a means to improve the mechanical performance of 316LVM coronary stents. Two different temperatures (1000 degrees C and 1150 degrees C) were explored for the thermal processing of the device. Acid pickling was done as a pre-annealing step to remove the debris and slag material attached to the stent after laser cutting. Post annealing operation involved the electrochemical polishing of the device which was also a parameter for assessment of the feasibility of the annealing process. Microstructural characterization, balloon expandability and tensile testing of the stents were performed to characterize the properties after thermal treatment. A fine grained austenitic structure with marked improvement in the % elongation (>40%) could be achieved after annealing the stents at 1000 degrees C. Balloon expandability tests of the stents annealed at 1000 degrees C indicated that the device was implantable. PMID- 17119278 TI - Endothelial cell attachment to the gamma irradiated small diameter polyurethane vascular grafts. AB - Previously we have fabricated the small diameter polyurethane (Pellethane 2363 80A, abbreviated PU) vascular grafts that were modified by epoxy-crosslinked gelatin (abbreviated gelatin) and an RGD-containing protein (abbreviated CBD-RGD) to facilitate the endothelial cell (EC) seeding on the surface. In this study, the biocompatibility of such surface after freeze-drying and gamma irradiation was evaluated. The contact angle of the irradiated PU dropped a little and the ESCA spectra revealed oxygen bonding. The increases in the amount of extractables as well as in the molecular weight distribution were observed. The mechanical properties decreased only slightly. The irradiated PU surface showed enhanced EC affinity that persisted after several months of storage. Gelatin, CBD-RGD (used with either gelatin or PU), and PU modified by gelatin and CBD-RGD all demonstrated higher EC affinity after freeze-drying and gamma irradiation (2.5 Mrad). The positive cellular effect remained after storage. Based on these results, freeze-drying followed by gamma irradiation at 2.5 Mrad is a proper way to process and store these vascular grafts. PMID- 17119279 TI - Rapid preparation of fresh frozen tissue-engineered bone sections for histological, histomorphological and histochemical analyses. AB - Fresh frozen sections are the best materials to assess tissue-engineered bone using cells/ceramic complexes. However, there are a lot of technical difficulties in obtaining serial sections suitable for microscopic examinations. Kawamoto et al. developed a method for the production of fresh frozen sections using new adhesive tape, and showed that sections were very useful for histological and histochemical studies. However, no study reported that the method was useful for tissue-engineered bone from histochemical and histomorphological points of view. This study aimed to determine the efficacy of fresh frozen sectioning for evaluating tissue-engineered bone. We revealed that fresh frozen sections retained the original morphology of tissue-engineered bone, and their biochemical characteristics. Therefore, rapid preparation of fresh frozen sections using adhesive tape is extremely useful for research of tissue-engineered bone, and serial sections can be assessed from both histomorphological and biochemical point of views. It is expected that this method will become a powerful tool in tissue-engineering of hard tissues. PMID- 17119280 TI - Failures of yttria-stabilised tetragonal zirconia: 52 retrieved ceramic femoral heads of total hip prostheses. AB - Fifty two ceramic femoral heads have been investigated after implantation. All heads had articulated against ultra high molecular weight polyethylene (UHMWPE) cups. Examination of the heads shows changes in sphericity and surface roughness due to the wear. The behaviour of these retrieved heads after clinical use confirms the problems derived from the use of yttria-stabilised tetragonal zirconia heads. In this paper, a failure mechanism for this type of ceramic is proposed. The slow yttria dissolution produces the transformation from tetragonal to monoclinic phase with a volume change. This fact provokes an important decrease in mechanical properties favouring wear. The yttria dissolution and the zirconia solid transformation were confirmed by X-ray diffraction. PMID- 17119281 TI - Mechanical properties of calcium sulphate/hydroxyapatite cement. AB - Setting times, volume after setting, injectability and hardness (at 37 degrees C in contact with Ringer's solution) were determined for cements made of mixtures of calcium sulphate hemihydrate (CS) and hydroxyapatite (HA) with a range of compositions. The purpose of these experiments was to determine the behaviour of a mixture that could be used as an injectable cement for orthopaedic applications, including spinal fusion. A suitable mixture consisted of 60% CS and 40% HA by mass; a slurry was made by mixing solid (36 g) with water (15 cm(3)). The slurry had initial and final setting times of 5.7+/-1.3 min and 19.6+/-0.7 min (mean +/- standard deviation), respectively. The hardness of the cement did not systematically increase or decrease in the 72 h following the final setting time. The volume of the cement was 99.8+/-0.4% of the volume of the initial slurry, i.e. there was negligible shrinkage on setting. It was able to withstand a pressure of 7.3+/-1.2 MPa, applied by a hemispherical indenter before the onset of permanent damage, indicating adequate strength for spinal fusion. PMID- 17119283 TI - Mechanisms of aluminum-induced neurodegeneration in animals: Implications for Alzheimer's disease. AB - For four decades the controversial question concerning a possible role for aluminum neurotoxicity in contributing to the pathogenesis of Alzheimer's disease has been debated, and studies by different investigators have yielded contradictory results. The lack of sensitivity to aluminum neurotoxicity in transgenic mouse models of Alzheimer's disease has not allowed the system to be used to explore important aspects of this toxicity. Rabbits are particularly sensitive to aluminum neurotoxicity and they develop severe neurological changes that are dependent on dose, age and route of administration. The most prominent feature induced by aluminum in rabbit brain is a neurofibrillary degeneration that shares some similarity with the neurofibrillary tangles found in Alzheimer's disease patients. In the present review we discuss data from our laboratory and others, on the effects of aluminum on behaviour, neurologic function and morphology, using aluminum administered to rabbits via different routes. Finally, we will examine data on the possible cellular mechanisms underlying aluminum neurotoxicity, and potential neuroprotective strategies against aluminum toxicity. PMID- 17119284 TI - Metals and Alzheimer's disease. AB - There is increasing evidence to support a role for both the amyloid beta-protein precursor (AbetaPP) and its proteolytic fragment, amyloid beta (Abeta), in metal ion homeostasis. Furthermore, metal ions such as zinc and copper can interact with both AbetaPP and Abeta to potentiate Alzheimer's disease by participating in the aggregation of these normal cellular proteins and in the generation of reactive oxygen species. In addition, metal ions may interact on several other AD related pathways, including those involved in neurofibrillary tangle formation, secretase cleavage of AbetaPP and proteolytic degradation of Abeta. As such, a dysregulation of metal ion homeostasis, as occurs with both aging and in AD, may foster an environment that can both precipitate and accelerate degenerative conditions such as AD. This offers a broad biochemical front for novel therapeutic interventions. PMID- 17119285 TI - The role of aluminum and copper on neuroinflammation and Alzheimer's disease. AB - Metals such as aluminum (Al), copper (Cu), zinc and iron have been implicated in the pathogenesis of Alzheimer's disease (AD). Because trace amounts of metals are present in the drinking water, there is a possibility for low-dose chronic exposure. Since the presence of Al and Cu in drinking water has been shown to adversely affect the progression of AD, these two metals may aggravate some of the events associated with the disease process. The main focus of this review will be on the effects of Al and Cu in initiating or propagating an inflammatory response within the aging brain. Since inflammatory events are reported to be upregulated in the AD brain, this may be one of the mechanisms by which the metals potentiate neurodegeneration. PMID- 17119286 TI - Aluminium and iron, but neither copper nor zinc, are key to the precipitation of beta-sheets of Abeta_{42} in senile plaque cores in Alzheimer's disease. AB - A number of metals including Fe(II)/Fe(III), Al(III), Zn(II) and Cu(II) are found co-localised with beta-sheets of Abeta_{42} in senile plaque cores in AD brain. We know neither why nor how the co-localisation takes place or, indeed, if it is entirely aberrant or partly protective. There are data from in vitro studies which may begin to explain some of these unanswered questions and in considering these I have summised that Al(III) and Fe(III)/Fe(II) are directly involved in the precipitation of beta-sheets of Abeta_{42} in senile plaque cores whereas the presence of Cu(II) and Zn(II) is adventitious. The co-deposition of Al(III), Fe(III) and beta-sheets of Abeta_{42} could act as a source of reactive oxygen species and begin to explain some of the oxidative damage found in the immediate vicinity of senile plaques. Whether such metal-Abeta_{42} synergisms are an integral part of the aetiology of AD remains to be confirmed. PMID- 17119287 TI - Aluminum and Alzheimer's disease: a new look. AB - Despite the circumstantial and sometimes equivocal support, the hypothetic involvement of aluminum (Al) in the etiology and pathogenesis of Alzheimer's disease (AD) has subsisted in neuroscience. There are very few other examples of scientific hypotheses on the pathogenesis of a disease that have been revisited so many times, once a new method that would allow a test of Al's accumulations in the brain of AD patients or a comparison between Al-induced and AD neuropathological signs has become available. Although objects of methodological controversies for scientists and oversimplification for lay spectators, several lines of evidence have strongly supported the involvement of Al as a secondary aggravating factor or risk factor in the pathogenesis of AD. We review evidence on the similarities and dissimilarities between Al-induced neurofibrillary degeneration and paired helical filaments from AD, the accumulation of Al in neurofibrillary tangles and senile plaques from AD, the neuropathological dissociation between AD and dialysis associated encephalopathy, and the epidemiological relations between Al in drinking water and the prevalence of AD. We also critically analyze the prospects of Al-amyloid cascade studies and other evolving lines of evidence that might shed insights into the link between Al and AD. The message between the lines of the following article is that the involvement of Al in the pathogenesis of AD should not be discarded, especially in these times when the amyloid dogma of AD etiology shows its myopia. PMID- 17119288 TI - Experimental approaches to assess metallotoxicity and ageing in models of Alzheimer's disease. AB - This review highlights advantages and disadvantages of experimental procedures (chemical, cellular, physiological, histochemical and epidemiological) that have been used to identify Alzheimer- and dementia-related targets for exogenous toxins, and discusses how neuronal function can be assessed experimentally, based on the evidence obtained for the neurotoxin aluminium. PMID- 17119289 TI - Mapping and characterization of iron compounds in Alzheimer's tissue. AB - Understanding the management of iron in the brain is of great importance in the study of neurodegeneration, where regional iron overload is frequently evident. A variety of approaches have been employed, from quantifying iron in various anatomical structures, to identifying genetic risk factors related to iron metabolism, and exploring chelation approaches to tackle iron overload in neurodegenerative disease. However, the ease with which iron can change valence state ensures that it is present in vivo in a wide variety of forms, both soluble and insoluble. Here, we review recent developments in approaches to locate and identify iron compounds in neurodegenerative tissue. In addition to complementary techniques that allow us to quantify and identify iron compounds using magnetometry, extraction, and electron microscopy, we are utilizing a powerful combined mapping/characterization approach with synchrotron X-rays. This has enabled the location and characterization of iron accumulations containing magnetite and ferritin in human Alzheimer's disease (AD) brain tissue sections in situ at micron-resolution. It is hoped that such approaches will contribute to our understanding of the role of unusual iron accumulations in disease pathogenesis, and optimise the potential to use brain iron as a clinical biomarker for early detection and diagnosis. PMID- 17119290 TI - Blood-brain barrier flux of aluminum, manganese, iron and other metals suspected to contribute to metal-induced neurodegeneration. AB - The etiology of many neurodegenerative diseases has been only partly attributed to acquired traits, suggesting environmental factors may also contribute. Metal dyshomeostasis causes or has been implicated in many neurodegenerative diseases. Metal flux across the blood-brain barrier (the primary route of brain metal uptake) and the choroid plexuses as well as sensory nerve metal uptake from the nasal cavity are reviewed. Transporters that have been described at the blood brain barrier are listed to illustrate the extensive possibilities for moving substances into and out of the brain. The controversial role of aluminum in Alzheimer's disease, evidence suggesting brain aluminum uptake by transferrin receptor mediated endocytosis and of aluminum citrate by system Xc;{-} and an organic anion transporter, and results suggesting transporter-mediated aluminum brain efflux are reviewed. The ability of manganese to produce a parkinsonism like syndrome, evidence suggesting manganese uptake by transferrin- and non transferrin-dependent mechanisms which may include store-operated calcium channels, and the lack of transporter-mediated manganese brain efflux, are discussed. The evidence for transferrin-dependent and independent mechanisms of brain iron uptake is presented. The copper transporters, ATP7A and ATP7B, and their roles in Menkes and Wilson's diseases, are summarized. Brain zinc uptake is facilitated by L- and D-histidine, but a transporter, if involved, has not been identified. Brain lead uptake may involve a non-energy-dependent process, store operated calcium channels, and/or an ATP-dependent calcium pump. Methyl mercury can form a complex with L-cysteine that mimics methionine, enabling its transport by the L system. The putative roles of zinc transporters, ZnT and Zip, in regulating brain zinc are discussed. Although brain uptake mechanisms for some metals have been identified, metal efflux from the brain has received little attention, preventing integration of all processes that contribute to brain metal concentrations. PMID- 17119291 TI - Heme binding to Amyloid-beta peptide: mechanistic role in Alzheimer's disease. AB - Genetic, biochemical, and immunological evidences support a mechanistic role for amyloid-beta (Abeta) peptide in the pathophysiology of Alzheimer's disease (AD). Abeta appears to trigger most of the disparate cytopathologies of AD (e.g. loss of iron homeostasis and mitochondrial complex IV), which may initiate synaptic dysfunction, hypometabolism, and memory loss. However, the molecular mechanism that links Abeta to the neurodegeneration of AD is not clear. We have provided evidence for heme's key role in the important cytopathologies of AD, hypothesizing a functional deficiency for heme in the brains of AD patients. The molecular link between beta and heme required to support this hypothesis was demonstrated by our discovery that heme binds with Abeta, forming a complex (Abeta-heme). Heme prevented the aggregation of Abeta by forming Abeta-heme, suggesting Abeta-heme may prevent Abeta aggregation in vivo. The downside, however, is that Abeta-heme is a peroxidase, which if not regulated might indiscriminately oxidize diverse biomolecules. Additionally, excessive production of Abeta in AD brain may bind to and restrict the bioavailability of regulatory heme, creating a condition of heme-deficiency. Regulatory heme regulates heme synthesis, iron homeostasis, specific signaling pathways, and intermediary metabolism. A novel model of Abeta-induced heme-deficiency leading to mitochondrial dysfunction, Abeta-heme peroxidase, and altered metabolic activity is presented. Genetic, nutritional, and toxicological factors that influence heme metabolism will be discussed in relevance to AD. PMID- 17119292 TI - HFE mutations and Alzheimer's disease. AB - An imbalance in brain iron status has been established in Alzheimer's Disease (AD). This iron imbalance can impact plaque formation, amyloid processing, and expression of and response to inflammatory agents. In a more general sense, a deregulation of iron homeostasis underlies the generation of reactive oxygen species leading to oxidative damage. Thus, loss of iron homeostasis can be central to the pathogenic events in AD. Recently a number of studies have begun to investigate the frequency of mutations in the HFE gene in AD. Mutations in the HFE gene occur more frequently in Caucasians than any other mutation. The two most common mutations of HFE are the C282Y (2% of the total population) and the H63D (9%. Mutations in this gene are associated with loss of iron homeostasis, alterations in inflammatory responses and in its most severe form, a clinical disorder known as Hemochromatosis. The C282Y mutation is more frequently associated with Hemochromatosis and the frequency of the H63D mutation is receiving increasing attention in neurodegenerative disorders. This review summarizes the data on HFE mutations in neurodegenerative disorders and what is known about HFE in the brain and the cell biology underlying the HFE mutation. PMID- 17119293 TI - Presenilin structure in mechanisms leading to Alzheimer's disease. AB - Molecular genetic studies of familial Alzheimer's disease by 1995 had clearly implicated three proteins as critical to Alzheimer's disease (AD), the amyloid beta protein precursor (AbetaPP) and the two homologous presenilins, PS-1 and PS 2. To account for the roles of these proteins in AD, we had proposed that as an early and critical step in the mechanisms that lead to AD, the PS on the surface of a brain cell engages in a specific receptor-ligand intercellular interaction with AbetaPP on the surface of a neighboring cell. This cell-cell interaction is required to trigger off a cascade of processes that lead to the production of amyloid-beta (Abeta) from AbetaPP, leading to AD. At about this time, however, many established AD researchers had obtained data that appeared to disagree with our proposed mechanism. Their immediate objections to our proposal were based on their conclusions that 1) The PS proteins were exclusively intracellular, and were not expressed at the cell surface, and 2) The topography of the PS proteins in intracellular membranes exhibits either 6 or 8-TM spanning domains, not 7. Here we discuss the evidence for the 6-TM, 7-TM, 8-TM and other models of PS topography and offer possibilities for the differences in interpretation of the various sets of data. We review the experimental demonstration of the cell surface expression and the 7-TM structure of PS, the functional consequences of this structure, and the findings that PS-1 and PS-2 are members of the superfamily of 7-TM heterotrimeric G-protein coupled receptors (GPCRs). PMID- 17119294 TI - The nucleation growth and reversibility of Amyloid-beta deposition in vivo. AB - The amyloid-beta (Abeta) peptide is a major constituent of the brain senile plaques that characterize Alzheimer's disease (AD). Converging observations led to the formulation of the amyloid hypothesis whereby the accumulation of soluble aggregates and insoluble Abeta deposits is the primary event in AD pathogenesis. Furthermore, the apoE4 isoform of apolipoprotein E, a major prevalent genetic risk factor of AD, is associated with increased Abeta deposition. To investigate the initial stages of the amyloid cascade in vivo and how this is affected by apoE4, we studied the effects of prolonged inhibition and subsequent reactivation of the Abeta-degrading enzyme, neprilysin, on aggregation and deposition of Abeta in apoE transgenic and control mice. The results revealed that Abeta deposition in vivo is initiated by aggregation of Abeta42, which is followed by reversible deposition of both Abeta42 and Abeta40, along with growth of the deposits, and by their subsequent irreversible fibrillization. The initiation of Abeta42 deposition is accelerated isoform-specifically by apoE4, whereas the growth and dissolution of the Abeta deposits as well as their fibrillization are similarly stimulated by the various apoE isoforms. Interestingly, Abeta deposition was associated with increased gliosis, which may reflect early pathological interactions of beta with the brain's parenchyma. PMID- 17119295 TI - Macronutrients, aluminium from drinking water and foods, and other metals in cognitive decline and dementia. AB - A possible role of the macronutrients and the basic elements of carbohydrates (glucose administration or depletion), proteins (amino acids such as tryptophan and tyrosine), and fat (unsaturated fatty acids) was recently proposed for age related changes of cognitive function, and the cognitive decline of degenerative (AD) or vascular origin. The availability and utilization of glucose has been implicated in cognitive function not only as a result of nutritional and systemic metabolic conditions, but also, although speculatively, as a crucial phase of the mechanism of action of molecules used as cognitive-enhancers. Furthermore, many lines of evidence have focused on the importance of oxidative stress mechanisms and free radical damage in AD pathogenesis. In addition, epidemiological studies have recently reported an association between alcohol and the incidence of AD and predementia syndromes. Foods with large amounts of aluminium-containing additives or aluminium from drinking water may affect the risk of developing AD, aluminium more likely acting as a cofactor somewhere in the cascade of events leading to the demented brain. A role for other metals in dementia have been speculated, given the encouraging results reported from studies on peripheral zinc concentrations, zinc supplementation, serum copper, either bound with ceruloplasmin or not, and iron metabolism in AD. Nonetheless, more data are needed to support a possible role of these metals in dementing diseases. Healthy diets, antioxidant supplements, and the prevention of nutritional deficiencies or exposure to foods and water with high content of metals could be considered the first line of defence against the development and progression of cognitive decline. PMID- 17119296 TI - Aluminum and other metals in Alzheimer's disease: a review of potential therapy with chelating agents. AB - Alzheimer's disease (AD) is characterized by the presence of neuritic plaques and neurofibrillary tangles in the brain. Although the causes of AD remain still unknown, it seems that certain environmental factors may be involved in the etiology and pathogenesis of the disease. While AD is associated with the abnormal aggregation of beta-amyloid protein in the brain, evidence shows that certain metals play a role in the precipitation and cytotoxicity of this protein. Among these metals, the potential role of aluminum as a possible ethiopathogenic factor in AD has been especially controversial. This review is mainly focused on the role of aluminum and metals such as copper and zinc in AD, as well as on metal chelator therapy as a potential treatment for AD. The effects of desferrioxamine and other Al chelating agents have been reviewed. The role of the metal chelator clioquinol in AD, which has been reported to reduce beta-amyloid plaques, presumably by chelation associated with copper and zinc, is also revised. Finally, the potential role of silicon in AD is also discussed. PMID- 17119298 TI - Plasticity in spatial neglect: recovery and rehabilitation. PMID- 17119299 TI - Impact of neglect on functional outcome after stroke: a review of methodological issues and recent research findings. AB - PURPOSE: This study provides an update on recent research findings concerning neglect and its impact on functional outcome. The review covers studies published during the past ten years. METHODS: A systematic review was carried out on reports drawn from electronic databases (MEDLINE and PSYCHLIT, January 1996 - August 2005) and identified from the lists of references in these reports. Unpublished reports, articles in other than the English language, and studies with non-human and non-adult subjects were excluded. The selection criteria were met by 26 articles. RESULTS: 15 of the 26 studies recruited heterogeneous patient groups (patients with right and left and/or unspecified lesions). The results from homogeneous groups (right hemisphere patients) were more consistent, emphasizing neglect as an independent predictor of functional outcome. Studies with homogeneous patient groups used consecutive series of patients, standardized measures of neglect, and a broader concept of functional outcome (both motor and cognitive items) than those with heterogeneous patient groups. Follow-ups longer than one year were very rare. CONCLUSIONS: Neglect has a significant negative impact on functional outcome, either as an independent predictive factor or in connection with other variables. The results, however, are inevitably affected by differences in patient samples and in the methods used in assessing neglect and functional outcome. Research focusing on homogeneous patient groups and especially on left hemisphere patients is needed. Neglect should be assessed with a standardized test battery rather than a single test, and functional outcome should be measured with scales consisting of cognitive, social and motor items. Also longer follow-ups are needed to verify the long-term functional outcome of neglect patients. PMID- 17119300 TI - Neglect and extinction: within and between sensory modalities. AB - PURPOSE: The interest in human conscious awareness has increasingly propelled the study of neglect, the most striking occurrence of an acquired lack of conscious experience of space. Neglect syndromes commonly arise after unilateral brain damage that spares primary sensory areas nonetheless leading to a lack of conscious stimulus perception. Because of the central role of vision in our everyday life and motor behaviour, most research on neglect has been carried out in the visual domain. Here, we suggest that a comprehensive perspective on neglect should examine in parallel evidence from all sensory modalities. METHODS: We critically reviewed relevant literature on neglect within and between sensory modalities. RESULTS: A number of studies have investigated manifestations of neglect in the tactile and auditory modalities, as well as in the chemical senses, supporting the idea that neglect can arise in various sensory modalities, either separately or concurrently. Moreover, studies on extinction (i.e., failure to report the contralesional stimulus only when this is delivered together with a concurrent one in the ipsilesional side), a deficit to some extent related to neglect, showed strong interactions between sensory modality for the conscious perception of stimuli and representation of space. CONCLUSIONS: Examining neglect and extinction by taking into account evidence from all sensory modalities in parallel can provide deeper comprehension of the neglect syndrome mechanisms and possibly more effective multi-sensory based rehabilitation approaches. PMID- 17119301 TI - Stimulus- and goal-driven biases of selective attention following unilateral brain damage: implications for rehabilitation of spatial neglect and extinction. AB - In this review we address the question of whether selective attentional mechanisms within the ipsilesional field are intact in unilateral lesion patients with spatial neglect and extinction. We consider how a lesion-induced bias in the neural representation of salience critically disrupts the integration of goal driven and stimulus-driven prioritization signals. This has important consequences for selectivity both within the 'impaired' contralesional field within the 'intact' ipsilesional field. Examples are drawn from the neuropsychological literature and recent experiments conducted within our own laboratory. The implications of ipsilesional spatial selection deficits for rehabilitation are discussed. PMID- 17119302 TI - Anosognosia for motor and sensory deficits after unilateral brain damage: a review. AB - PURPOSE: The syndrome of unawareness (anosognosia) for sensory and motor neurological deficits (hemiplegia, hemianaesthesia, and hemianopia), contralateral to the side of a hemispheric lesion, is reviewed. CONTENT: Main topics include: basic historical facts; the types of patient's interview and specific questions used to reveal the deficits; the clinical patterns of presentation; the associations and dissociations of the different anosognosic manifestations, and their relationships with associated disorders of sensory, memory, and executive-intellectual functions; the hemispheric asymmetry of anosognosia, that, as the syndrome of unilateral spatial neglect, is more frequent and severe after damage to the right cerebral hemisphere; the relationships between spatial neglect and the anosognosias, and their neural correlates; the effects of lateralized sensory stimulations on defective awareness of neurological impairments. CONCLUSIONS: The argument is made that anosognosia for sensory and motor neurological deficits should be considered as a multi-component syndrome, including a number of specific disorders that are due to the impairment of discrete monitoring systems, specific for the different supervised functions. The putative causal role of associated deficits of other parts of the sensory-motor or cognitive (e.g., memory, general intelligence) system is critically discussed. These specific control processes may be physically implemented in brain areas anatomically (and functionally) close to those subserving the monitored function. PMID- 17119303 TI - What do eye-fixation patterns tell us about unilateral spatial neglect? AB - PURPOSE: Eye-fixation patterns, which include ocular searching and fixation, may change with tasks, stimuli, and instructions. This article reviews our studies over 18 years on eye-fixation patterns of neglect patients and aims to elucidate the visuospatial processing of unilateral spatial neglect. METHODS: We recorded eye-fixation patterns when patients with neglect bisected a line in various conditions. RESULTS: Patients with neglect rarely searched to the left side when bisecting a line of the ordinary length (e.g., 200 mm). They persisted in fixating a right-side point, at which they later marked the subjective midpoint. They made no effective comparison between the leftward and rightward extents not only for a whole line but also for its explored right segment. Where they 'favored' to fixate as the subjective midpoint depended strongly upon the location of the right endpoint in space. Their representational image of a line was also estimated with modified line bisection tasks performed on a touch-panel display. CONCLUSIONS: For patients with neglect, the representational image of a line may be formed on the basis of the attended segment between the right endpoint and the favored point of fixation. The line bisection task, if combined with recording of eye-fixation, would further contribute to elucidation of the mechanisms underlying neglect. PMID- 17119304 TI - A battery of tests for the quantitative assessment of unilateral neglect. AB - PURPOSE: The lack of agreement regarding assessment methods is responsible for the variability in the reported rate of occurrence of unilateral neglect (UN) after stroke. In addition, dissociations have been reported between performance on traditional paper-and-pencil tests and UN in everyday life situations. METHODS: In this paper, we present the validation studies of a quantitative test battery for UN, including paper-and-pencil tests, an assessment of personal neglect, extinction, and anosognosia, and a behavioural assessment, the Catherine Bergego Scale (CBS). The battery was given to healthy subjects (n=456-476) and to patients with subacute stroke, either of the right or the left hemisphere. RESULTS: In healthy subjects, a significant effect of age, education duration and acting hand was found in several tasks. In patients with right hemisphere stroke, the most sensitive paper and pencil measure was the starting point in the cancellation task. The whole battery was more sensitive than any single test alone. An important finding was that behavioural assessment was more sensitive than any other single test. Neglect was two to four times less frequent, but also less severe and less consistent after left hemisphere stroke. CONCLUSION: Assessment of UN should rely on a battery of quantitative and standardised tests. Some patients may show clinically significant UN in everyday life while obtaining a normal performance on paper-and-pencil measures. This underlines the necessity to use a behavioural assessment of UN. PMID- 17119305 TI - Spatial and non-spatial attention deficits in neurodegenerative diseases: assessment based on Bundesen's theory of visual attention (TVA). AB - PURPOSE: The aim was to present evidence that, similarly as in neglect, a combined pattern of spatial and non-spatial deficits of visual attention can also be typically observed in patients suffering from neurodegenerative disorders. METHOD: Whole and partial report of brief letter arrays, based on Bundesen's 'theory of visual attention' (TVA), was applied in patients suffering from Huntington's disease (HD), mild cognitive impairment (MCI), or Alzheimer's disease (AD). TVA-based parameter estimates were derived reflecting (a) perceptual processing speed and visual working memory storage capacity as non spatial aspects of visual attention (determined by whole report performance), and (b) spatial attentional weighting (determined by partial report performance). RESULTS: Processing speed was severely slowed in HD, and also reduced, although to a lesser degree, in MCI and AD patients. In HD and AD patients, but not in MCI patients, a strong leftward bias of spatial attention was observed. CONCLUSION: Neglect and neurodegenerative diseases both involve a similar constellation of non-spatial and spatial deficits of visual attention. Therefore, by using TVA based measurement, results from both fields of research may fruitfully inform each other in future studies, thus improving our understanding of the interaction of spatial and non-spatial attention deficits and its behavioral consequences. PMID- 17119306 TI - Construction and psychometric properties of a novel test for body representational neglect (Vest Test). AB - PURPOSE: Multimodal spatial neglect manifests itself also in nonvisual modalities such as audition, touch and body representation. Yet, quantitative tests for the diagnosis of nonvisual neglect are still quite rare. The purpose of the present paper was to develop and evaluate a novel, simple and sensitive test for the assessment of body representational neglect (BRN) in patients with left or right cerebral hemispheric lesions. METHODS: The vest test covers the front part of the trunk. The blindfolded subject wears the vest and is instructed to pick up all objects from the 24 pockets of the vest (12 on each side) as quickly as possible using the ipsilesional, nonparetic hand. Two samples of healthy control subjects (each N=25) using either their left or their right hand performed the test in identical way to obtain normative data for patients searching with their left hand (i.e. left hemisphere stroke patients) versus their right hand (i.e. right hemisphere stroke patients). The test can be performed within 5~minutes, even with aphasic or apractic stroke patients. RESULTS: Psychometric evaluations in a sample of 50 patients with unilateral stroke (25 leftsided, 25 rightsided) show high objectivity, high internal consistency (Cronbach's alpha=0.96), good retest reliability (0.79 after 1 week in neglect patients) and good validity as compared with two other measures of BRN or multimodal neglect. Patient examples show that BRN as assessed with the vest test allows the detection of qualitatively and quantitatively different patterns of BRN, and shows double dissociations from visual neglect and from apraxia in left hemisphere stroke patients. Details of the test including instructions and cut-off values are given for users in the appendix of this article. CONCLUSIONS: In conclusion, the vest test is a sensitive, quick and reliable test for BRN which complements the assessment of visuo- and audiospatial neglect and allows to measure recovery (spontaneous or treatment-induced) in patients with BRN. Furthermore, it can help to improve our knowledge about the multisensory coding of our body and the surrounding space in the human brain. PMID- 17119307 TI - Postural disorders and spatial neglect in stroke patients: a strong association. AB - PURPOSE: In this paper we analyse the arguments for a strong association between spatial neglect and postural disorders and attempt to better understand the mechanisms which underlie that. METHODS: We first provide a general overview of the available tools for a rational assessment of postural control in a clinical context. We then analyse the arguments in favour of a close relationship, although not necessarily causal, between spatial neglect and: 1) body orientation with respect to gravity (including verticality perception i.e. the visual vertical, the haptic vertical, and the postural vertical); 2) body stabilisation with respect to the base of support; 3) posturographic features of stroke patients; 4) and finally their postural disability in daily life. This second part of the paper is based both on the literature review and on results of our current research. RESULTS: Neglect patients show a dramatic postural disability, due both to problems in body orientation with respect to gravity and to problems in body stabilisation. It might be that these problems are partly caused by a neglect phenomenon bearing on graviceptive (somaesthetic > vestibular) and visual information serving postural control. This could correspond to a kind of postural neglect involving both the bodily and nonbodily domains of spatial neglect. The existence of distorsion(s) in the body scheme are also probably involved, especially to explain the weight-bearing asymmetry in standing, and probably an impaired multisegmental postural coordination leading to an impaired body stabilisation. CONCLUSION: The present paper explains why neglect patients show longer/worse recovery of postural-walking autonomy than other stroke patients. PMID- 17119308 TI - Development of a rehabilitative program for unilateral neglect. AB - PURPOSE: The aim of the present paper is to review several studies which assessed the validity of a visuo-spatial training for the rehabilitation of neglect patients. In addition two peripheral stimulations (TENS and Optokinetic Stimulation) have been studied to assess the improvements of neglect disorders when used in combination with the visuo-spatial training. Also we analyzed the potential effect of training for attention on neglect and, viceversa, the effect of visuo-spatial training on attentional impairments. METHODS: the goals have been investigated by both group studies and descriptions of single cases. RESULTS: The visuo-spatial training produced significant improvements on the performance of neglect patients which generalized to every day living situations: the results showed to be stable over time and had positive effects on a variety of other neurological impairments. It was also shown that the improvements are confined to tasks involving spatial exploration of extrapersonal space, but did not extend to other neglect disorders, such as representational and personal neglect. The use of peripheral stimulations, at variance with other studies in the literature, did not add any advantage as compared to the improvements produced by the visuo-spatial training. No transfer between training for neglect and attention was observed. CONCLUSIONS: the present review pointed out that neglect disorders can be improved in a clinically meaningful way: the studies described also showed some limitations and proposed the need of further researches in order to extend the improvements to several other aspects of the neglect syndrome. PMID- 17119309 TI - Neglect and prism adaptation: a new therapeutic tool for spatial cognition disorders. AB - PURPOSE: A large proportion of right-hemisphere stroke patients show unilateral neglect, a neurological deficit of perception, attention, representation, and/or performing actions within their left-sided space, inducing many functional debilitating effects on everyday life, and responsible for poor functional recovery and ability to benefit from treatment. This spatial cognition disorder affects the orientation of behaviour with a shift of proprioceptive representations toward the lesion side. METHODS: This shift can be reduced after a prism adaptation period to a right lateral displacement of visual field (induced by a simple target-pointing task with base-left wedge prisms). The modification of visuo-motor or sensory-motor correspondences induced by prism adaptation involves improvement of different symptoms of neglect. RESULTS: Classical visuo-motor tests could be improved for at least 2h after adaptation, but also non-motor and non-visual tasks. In addition, cross-modal effects have been described (tactile extinction and dichotic listening), mental imagery tasks (geographic map, number bisection) and even visuo-constructive disorders. These cognitive effects are shown to result from indirect bottom-up effects of the deeper, adaptive realignment component of the reaction to prisms. Lesion studies and functional imaging data evoke a cerebello-cortical network in which each structure plays a specific role and not all structures are crucial for adaptation ability. CONCLUSIONS: These cognitive effects of prism adaptation suggest that prism adaptation does not act specifically on the ipsilesional bias characteristic of unilateral neglect but rehabilitates more generally the visuo spatial functions attributed to the right cortical hemisphere. These results reinforce the idea that the process of prism adaptation may activate brain functions related to multisensory integration and higher spatial representations and show a generalization at a functional level. Prism adaptation therefore appears as a new powerful therapeutic tool for spatial cognition disorders. PMID- 17119310 TI - Repetitive optokinetic stimulation induces lasting recovery from visual neglect. AB - PURPOSE: To evaluate whether repetitive optokinetic stimulation with active pursuit eye movements leads to substantial and greater recovery from visual neglect as compared to conventional visual scanning training. METHODS: Two groups of five patients with leftsided hemineglect were consecutively collected and matched for clinical and demographic variables as well as neglect severity. One group received five treatment sessions of repetitive optokinetic stimulation (R OKS) within one week, while the other group received the same amount of conventional visual scanning training (VST) using identical visual stimuli and setup. All patients were treated in a single-subject baseline design with treatment-free intervals before (14 days) and after specific neglect therapy (14 days). Dependent variables were the improvements in digit cancellation, visuoperceptual and visuomotor line bisection and visual size distortion during treatment. The transfer of treatment effects was assessed by a paragraph reading test. RESULTS: The results showed superior effects of OKS treatment in all five patients which generalized across all tasks administered and remained stable at follow-up. In contrast, no significant improvements were obtained after VST training in any of these tasks, except in line bisection. CONCLUSION: We conclude that the presentation of moving visual stimulus displays with active smooth pursuit eye movements can be more efficient than conventional visual scanning training using static visual displays. PMID- 17119311 TI - Alertness-training in neglect: behavioral and imaging results. AB - PURPOSE: It has been proposed that the right hemisphere alerting network co activates, either directly or via the brainstem, the spatial attention system in the parietal cortex. The observation that measures of impaired alertness and sustained attention can be used to predict the outcome of neglect might suggest such a relationship, too. The aim of the present study was to investigate the effects of alertness training on hemispatial neglect. METHOD: A three-week computerised alertness training was applied to patients with chronic (> 3 months) stable visuospatial hemineglect. Training effects were investigated both in a single case and in a group of 7 patients by means of neuropsychological tests and functional magnetic resonance imaging (fMRI). RESULTS: After the training, the patients showed a significant improvement in a neglect test battery above any natural fluctuation during a three-week baseline phase. Improvements in the neglect tasks were accompanied by an increase of both right and left hemisphere frontal, anterior cingulate and superior parietal activation, areas known to be associated with both alertness and spatial attention. Four weeks after the end of the training, the patients' neglect test performance had mostly returned to baseline. Despite decreases of activation in some of the initially reactivated areas, increases in neural activity bilaterally in frontal areas, in the right anterior cingulate cortex, the right angular gyrus and in the left temporoparietal cortex remained. An Optokinetic Stimulation Training (OKS) in a control group of another 7 neglect patients led to comparable behavioral results. After the training, however, there was a reactivation mainly in posterior parts of both hemispheres suggesting training specific functional reorganization. CONCLUSION: The limited stability of the behavioral and reactivation results over time demonstrates that a three-week alertness or OKS training alone does not result in long lasting behavioral improvements and stable reactivation patterns in every patient. We rather suggest that combining alertness and spatial attention oriented training procedures might lead to a more stable amelioration of neglect symptoms. PMID- 17119312 TI - Using limb movements to improve spatial neglect: the role of functional electrical stimulation. AB - PURPOSE: Spatial neglect is common after right-hemisphere stroke and has proven resilient to a number of therapeutic interventions. Both active and experimenter induced passive movements of the left limb in left hemispace have been shown to ameliorate neglect in subsets of patients by improving performance on tasks requiring attention to the left side of space. However, the high incidence of contralesional hemiparesis and poor motor recovery in neglect makes active limb movement therapies applicable to only a small subset of patients. The purpose of our studies was to investigate the effects of passive movements of the left hand by functional electrical stimulation (FES), a common and portable motor rehabilitation technique, on performance in a visual scanning task. METHODS: The effect of FES-induced passive movement on target detection in a visual scanning task was compared to no movement and active movement conditions and also investigated in scanning tasks in both near and far space. RESULTS: Passive limb movement effects in neglect were variable across and within studies, reference spaces, and individuals, with a subset of positive responders differing from non responders in regard to constructional deficits and lesion location. CONCLUSIONS: The potential viability of FES as a therapy for neglect deserves further investigation and directions for future research in this area are discussed. PMID- 17119313 TI - The need for randomised treatment studies in neglect research. AB - PURPOSE: The aim was to review the methodological quality of trials to evaluate rehabilitation for spatial neglect and to determine the overall effectiveness of interventions METHODS: A systematic literature review and meta-analysis were conducted of trials completed by 2005. Trials identified were independently assessed for methodological quality by two reviewers. Outcomes were analysed as the standardised mean difference and 95% confidence intervals with random effects models. RESULTS: 25 trials of neglect rehabilitation were identified, 12 randomised controlled trials and 13 controlled clinical trials. The methodological quality was generally poor with only 4 trials achieving an A rating, i.e. low risk of selection bias. The immediate effect of cognitive rehabilitation on disability was small, 0.26 [-0.16, 0.67] and neither this nor the persisting effect 0.61 [-0.42, 1.63] was statistically significant. The most frequently used standardised neglect test (number of single letters correctly cancelled) favoured the experimental group 0.58 [0.10, 1.05] but was not significant. When cancellation errors were measured there was a small immediate effect favouring the experimental group, of borderline statistical significance, 0.65 [-1.28, -0.01] p=0.05, and a significant persisting effect -0.76 [-1.39, 0.13] p=0.02. Cognitive rehabilitation also significantly improved immediate (p=001) and persisting (p=0.02) line bisection performance but these findings are based on only four and one study respectively. CONCLUSIONS: The quality of trials identified was poor. Analysis of randomised controlled trials showed some evidence of an effect of intervention on measures of impairment. There was no evidence to support the effects of intervention on measures of disability. Further trials must use methods that reduce bias, have adequate statistical power, and include valid disability outcome measures. PMID- 17119314 TI - Prism adaptation first among equals in alleviating left neglect: a review. AB - PURPOSE: The current paper was designed to provide a critical overview on the different methods proposed for the rehabilitation of left spatial neglect. METHODS: On the basis of a previous systematic review of the literature, we analyzed all articles available aiming at reducing left spatial neglect which included a long term functional assessment. RESULTS: The aim of most early rehabilitation approaches, such as visuo-scanning training, was to re-orient visual scanning toward the neglected side. This review confirmed the utility of this method for rehabilitation purposes. More recent - theory driven - procedures, also based on a training approach, include limb activation, mental imagery training and video-feedback training. Although there is ground for optimism, the functional effectiveness of these methods still relies on few single-case studies. Newer methods have tried to stimulate automatic orientation of gaze or attention towards neglected space in a bottom-up fashion. Sensory stimulations can remove most of the classical signs of left neglect but their effects are short-lived. Such stimulations are not functionally relevant for rehabilitation except for trunk rotation or repeated neck muscle vibrations if they are associated with an extensive training program. A more promising intervention is prism adaptation given the growing evidence of relatively long term functional gains from comparatively short term usage. CONCLUSION: Overall, there is now evidence for several clinically relevant long-term benefits in the case of visual scanning training, mental imagery training, video feedback training, neck muscle vibration and trunk rotation if associated with visual scanning training and prism adaptation. However, the amount of evidence is still limited to a small number of relevant published articles and it is mandatory to continue the research in this field. In this review, the possible routes for new rehabilitation procedures are discussed on the basis of the actual knowledge regarding the neuro-cognitive mechanisms underlying the therapeutic effect of prism adaptation. PMID- 17119315 TI - Simulating unilateral neglect in normals: myth or reality? AB - Hemispatial neglect is a neurological deficit of perception, attention, representation, and/or performing actions within the left-sided space. The condition also produces many functional debilitating effects on everyday life, and is associated with poor functional recovery and inability to benefit from treatment. Numerous methods of rehabilitation (sensory stimulations or active training) have been proposed to alleviate neglect condition. It has been recently shown that visuo-manual adaptation to rightward optical shift leads to profound and enduring improvements of neglect symptoms. Based on the different methods commonly used in the rehabilitation of neglect, several techniques have been employed to simulate neglect symptoms in healthy subjects with a view to better understand the physiopathology of neglect. The present paper reviews studies of neglect-like behaviour in healthy individuals and in particular the use of prism adaptation as a procedure for simulating various symptoms of clinical neglect in normals. Neglect-like symptoms following prism adaptation offer insights as to the mechanisms of spatial neglect and provide an understanding of the interaction between low level sensorimotor processes and spatial cognition. Implications for the functional mechanisms and the anatomical substrates of prism adaptation are discussed in terms of inter-sensory plasticity and sensorimotor coordination and the way these may affect higher-level representations of space. PMID- 17119316 TI - Virtual reality applications for the remapping of space in neglect patients. AB - PURPOSE: The aims of the present article were the following: (i) to provide some evidence of the potential of virtual reality (VR) for the assessment, training and recovery of hemispatial neglect; (ii) to present data from our laboratory which seem to confirm that the clinical manifestation of neglect can be improved by using VR techniques; and (iii) to ascertain the neural bases of this improvement. METHODS: We used a VR device (DataGlove) interfaced with a specially designed computer program which allowed neglect patients to reach and grasp a real object while simultaneously observing the grasping of a virtual object located within a virtual environment by a virtual hand. The virtual hand was commanded in real time by their real hand. RESULTS: After a period of training, hemispatial neglect patients coded the visual stimuli within the neglected space in an identical fashion as those presented within the preserved portions of space. However it was also found that only patients with lesions that spared the inferior parietal/superior temporal regions were able to benefit from the virtual reality training. CONCLUSIONS: It was concluded that using VR it is possible to re-create links between the affected and the nonaffected space in neglect patients. Furthermore, that specific regions may play a crucial role in the recovery of space that underlies the improvement of neglect patients when trained with virtual reality. The implications of these results for determining the neural bases of a higher order attentional and/or spatial representation, and for the treatment of patients with unilateral neglect are discussed. PMID- 17119317 TI - Exogenous lipid pneumonia. PMID- 17119318 TI - Differential gene expression during development in two oligodendroglial cell lines overexpressing transferrin: a cDNA array analysis. AB - In the central nervous system, transferrin (Tf) is produced by oligodendroglial cells (OLGcs) and is essential for their development. Recently, using the complete cDNA of the human Tf gene, we obtained clones overexpressing Tf in two OLGc lines, N19 and N20.1, which represent different stages of differentiation. We showed that the overexpression of this glycoprotein promotes the maturation and myelinogenic capacity of both cell lines. In this work, using cDNA array technology, we examined changes induced by Tf in 1,176 genes. We found 41 genes differentially expressed in both cell lines, all of them involved in OLGc development. In the less mature cells (N19) overexpressing Tf, there was a significant increase in key enzymes of neurosteroid metabolism, such as cholesterol side chain cleavage cytochrome P450, 3beta-hydroxysteroid dehydrogenase and 5alpha-reductase type 1. In the more mature cell line (N20.1), Tf overexpression produced an induction of several mRNAs of the GABA(A) receptor subunits, of thyroid hormone receptors and of proteins involved in axon-glia interactions such as F3/contactin. In addition, in both cell lines, Tf overexpression induced an increase in the expression of different isoforms of transforming growth factor beta receptors and in several genes related to mitochondrial function and to complex lipid metabolism, crucial steps in myelin synthesis. Differentiation produced by Tf in both cell lines seems to occur by modulation of different genes depending on the maturational stage of the cells. Our findings provide new insights into the molecular basis of OLGc differentiation and on the role played by Tf in this process. PMID- 17119319 TI - Involvement of c-Jun-JNK pathways in the regulation of programmed cell death of developing chick embryo spinal cord motoneurons. AB - Key features of developmentally regulated programmed cell death (PCD) have been described for the first time in the chick nervous system. JNK/c-Jun pathway was involved in early events determining normal and pathological neuronal death as shown in experimental models. In the chick embryo, PCD of motoneurons (MNs) in ovo occurs within a well-defined temporal window and can be subjected to experimental manipulation. Taking advantage of this in vivo system, we explored the role of c-Jun and JNK pathway in the regulation of PCD in MNs. By using specific antibodies against phospho-c-Jun (Ser 63, 73) and JNK we demonstrated that before MNs acquire apoptotic phenotype there is an increase in c-Jun. Blockage of neuromuscular activity by the GABA agonist muscimol reduces PCD and diminishes c-Jun immunoreactivity in MNs. Extensive induction of PCD, either due to injection of beta-bungarotoxin or limb bud removal, is also preceded by an increase in c-Jun immunoreactivity that is also associated with upregulation of phospho-c-Jun and JNK. Translocation of JNK from cytoplasm to MN nuclei was also detected. After acute application of beta-bungarotoxin, which is a strong apoptotic stimulus for MNs, c-Jun phosphorylation occurs on serine 73, whereas serine 63 is the main site for c-Jun phosphorylation after limb bud removal. These results demonstrated that the JNK/c-Jun pathway is involved in the decision phase of normal and induced apoptosis in MNs. Pharmacological interventions involving this pathway should be explored as a potential therapeutic target for promoting MN survival. PMID- 17119320 TI - Effects of hyperbaric exposure with high oxygen concentration on the physical activity of developing rats. AB - The effects of hyperbaric exposure with high oxygen concentration on the physical activity of developing male rats were investigated. Five-week-old male rats were exposed to an atmospheric pressure of 1.25 with an oxygen concentration of 36.0% for 12 h (7.00-19.00 h) and exercised voluntarily for 12 h (19.00-7.00 h) daily for 8 weeks. The voluntary running activities were compared with those in age matched rats without hyperbaric exposure. In addition, the properties of the soleus and plantaris muscle fibers and their spinal motoneurons were examined. The voluntary running activities of rats with or without hyperbaric exposure increased during development. However, the mean voluntary running activities were higher in rats with hyperbaric exposure (7,104 m/day) than in those without hyperbaric exposure (4,932 m/day). The oxidative capacities of the soleus and plantaris muscle fibers and their spinal motoneurons increased following hyperbaric exposure. It is suggested that adaptations of neuromuscular units to hyperbaric exposure with high oxygen concentration enhance the metabolism, and thus, the function of neuromuscular units is promoted. PMID- 17119321 TI - The role of XBtg2 in Xenopus neural development. AB - In early neural development, active cell proliferation and apoptosis take place concurrent with cell differentiation, but how these processes are coordinated remains unclear. In this study, we characterized the role of XBtg2 in Xenopus neural development. XBtg2 transcripts were detected at the edge of the anterolateral neural plate and the neural crest region at the midneurula stage, and in eyes and in part of the neural tube at the tailbud stage. Translational inhibition of XBtg2 affected anterior neural development and impaired eye formation. XBtg2 depletion altered the expression patterns of the early neural genes, Zic3 and SoxD, at the midneurula stage, but not at the early neurula stage. At the midneurula stage, XBtg2-depleted embryos exhibited a marked decrease in the expression of anterior neural genes, En2, Otx2, and Rx1, without any changes in neural crest genes, Slug and Snail, or an epidermal gene, XK81. These results suggest that XBtg2 is required for the differentiation of the anterior neural plate from the midneurula stage, but not for the specification of the fate and patterning of the neural plate. XBtg2-depleted embryos also exhibited an increase in both proliferation and apoptosis in the anterior neural plate; however, the altered expression patterns of neural markers were not reversed by inhibition of either the cell cycle or apoptosis. Based on these data, we propose that XBtg2 plays an essential role in the anterior neural development, by regulating neural cell differentiation, and, independently, cell proliferation and survival. PMID- 17119322 TI - Early and long-term engraftment after autologous peripheral stem cell transplantation in acute myeloid leukemia patients. AB - This study aimed to identify which subset of CD34+ cells might be the most predictive of early and long-term hematopoietic recovery following autologous peripheral blood stem cell (PBSC) transplantation (PBSCT) in adult acute myeloid leukemia (AML) patients. The relationships between the number of 'mature' subsets of CD34+ cells (CD34+/CD33+, CD34+/CD38+, CD34+/DR+ and CD34+/CD90-) and 'immature' subsets of CD34+ cells (CD34+/CD33-, CD34+/CD38-, CD34+/DR- and CD34+/CD90+) and early and long-term hemoglobin, neutrophil and platelet counts were studied in a homogeneous series (for disease, pre-transplant chemotherapy, mobilization chemotherapy, conditioning regimen) of 26 AML patients after autologous PBSCT. Cell counts were performed before and after cryopreservation, but only after thawing were the cell counts used for correlation with early and long-term engraftment. The number of CD34+/CD38- cells infused correlated with the neutrophil (r = 0.88, p < 0.005) and platelet counts (r = 0.67, p < 0.05) at 12 months after PBSCT. This correlation was better than that for the total CD34+ cell dose at 12 months (r = 0.36, p = 0.09 for neutrophil count and r = 0.48, p = 0.06 for platelets count). The number of CD34+/CD90+ cells was also correlated with the platelet counts at 6 (r = 0.70, p < 0.05) and 12 months (r = 0.80, p = 0.005) after PBSCT. This correlation was better than the total dose of CD34+ cells at 6 (r = 0.31, p = 0.3) and 12 months (r = 0.48, p = 0.06) for the platelet counts. CD34+ subset analysis suggests that for early engraftment the total number of CD34+ cells infused is more strongly correlated than the CD34+ subsets, whereas the CD34+/CD38- and CD34+/CD90+ subsets may be associated with sustained long-term neutrophil and platelet engraftment. These findings may help to predict the repopulating capacity of PBSCs in AML patients after autologous PBSCT, especially when a relatively low number of CD34+ cells is infused. PMID- 17119323 TI - Does hepcidin affect erythropoiesis in hemodialysis patients? AB - INTRODUCTION: Prohepcidin is the precursor of hepcidin, a liver-derived peptide involved in iron metabolism by blocking its intestinal absorption and its release by the reticuloendothelial system. Iron overload and inflammation increase hepcidin expression, whereas anemia and hypoxia suppress it. In the present study prohepcidin levels were determined in the serum of hemodialysis (HD) patients and its correlations with iron metabolism markers, C-reactive protein (CRP) and hematocrit (Hct) were assessed. PATIENTS AND METHODS: Forty-sixHD patients and 22 healthy volunteers were enrolled in the study. Hct, serum prohepcidin, CRP, iron, ferritin, transferrin saturation and transferrin receptors were measured. The weekly erythropoietin dose, last-month intravenous iron dose and the patients' demographics were recorded. RESULTS: In comparison to the healthy volunteers, the HD patients had higher serum ferritin, transferrin receptors and CRP, lower serum iron and similar transferrin saturation and prohepcidin levels. In the patient group prohepcidin levels were negatively correlated with Hct but not with any other of the examined parameters. Multiple linear regression analysis considering age, inflammation, iron adequacy, erythropoietin dose and prohepcidin levels revealed that prohepcidin was the predominant determinant of Hct. CONCLUSIONS: Taking into account the low Hct levels in the HD patients of our study, it seems plausible that the prohepcidin levels assessed in this group are inappropriately high. These functionally high prohepcidin levels may be associated with the factors that inhibit erythropoiesis in HD patients. On the other hand, the absence of other expected correlations indicates that further studies are needed in order to definitely clarify this aspect. PMID- 17119324 TI - Neutropenia due to parvovirus B19 infections in patients with paroxysmal nocturnal hemoglobinuria. Blood transfusion and natural infection cases. AB - Two patients with paroxysmal nocturnal hemoglobinuria (PNH) and parvovirus B19 (PVB19) infection are reported. One was infected by PVB19-contaminated blood transfusion, whereas the other had become infected naturally. Both patients completely recovered after transient pancytopenia, but showed severe and intermediate neutropenia (0.216 and 0.768 x 10(9)/l, respectively) at the nadirs of aplastic crises. In the literature, PNH cases also showed neutropenia (0 and 0.54 x 10(9)/l) at aplastic crises. When patients with PNH are infected by PVB19, they may show severe neutropenia and mild thrombocytopenia in addition to severe reticulocytopenia. Therefore, these patients might have to be treated with granulocyte colony-stimulating factor to be able to recover from severe neutropenia. PMID- 17119325 TI - Soluble transferrin receptor-1 levels in mice do not affect iron absorption. AB - Soluble transferrin receptor-1 (sTfR1) concentrations are increased in the plasma under two conditions that are associated with increased iron absorption, i.e. iron deficiency and increased erythropoiesis. To determine the possible role of sTfR1 as a signaling mechanism for iron absorption, a hydrodynamic gene transfer technique was established to express transfected plasmid constructs of human sTfR1 (hsTfR1) and murine sTfR1 (msTfR1) from the livers of C57BL/6 mice. Iron absorption, serum iron levels and hepcidin expression were then measured. The hydrodynamic gene transfer technique proved to be an effective approach to achieving sustained expression of sTfR1 in mice. Although expression of high levels of sTfR1 significantly increased serum iron levels, repeated experiments showed that neither hsTfR1 nor msTfR1 had any effect on iron absorption or hepcidin mRNA expression levels. Thus, despite its attractiveness as a potential modifier of iron absorption, sTfR1 levels do not exert a regulatory effect on iron absorption. PMID- 17119326 TI - Bortezomib and dexamethasone in previously untreated multiple myeloma associated with renal failure and reversal of renal failure. AB - Multiple myeloma (MM) associated with renal failure carries a worse prognosis when compared with MM without renal failure. Bortezomib, a reversible proteosome inhibitor, is a new drug indicated for the treatment of refractory or relapsed myeloma. Published data on the use of bortezomib in patients with myeloma and renal failure are few. We report our experience with bortezomib and dexamethasone in 3 previously untreated and 1 relapsed patient with MM and renal failure. All patients achieved rapid improvement in their renal function as measured by serum creatinine levels with only 1-2 cycles of bortezomib (+/- dexamethasone), 3 of 4 patients had a near complete response and 1 patient had a partial response. The rapid reversal of renal dysfunction with bortezomib (+/- dexamethasone) treatment may be an effective strategy to prevent end stage renal failure in MM, thereby improving the morbidity and mortality in this otherwise poor prognosis subset of patients with myeloma. PMID- 17119327 TI - Variable efficacy of recombinant human erythropoietin in anemic pregnant women with different forms of heterozygous hemoglobinopathy. AB - OBJECTIVE: The aim of this study was to determine the response to recombinant human erythropoietin (rhEPO) in anemic pregnant women with heterozygous hemoglobinopathies. METHODS: A prospective study including 19 consecutive pregnant women with anemia and heterozygous hemoglobinopathy was performed. Treatment was divided into two phases: the initial low-dose phase and the subsequent high-rhEPO phase. In the initial phase, 3 x 10,000 U of rhEPO was administered with intravenous iron sucrose. In patients showing a poor response (Hb increase <1 g/dl) to low-dose rhEPO, the rhEPO dose was increased to 20,000 U per treatment in the subsequent phase. RESULTS: All patients showed stimulation of erythropoiesis as evidenced by an increase in hemoglobin. In 13 patients, a good response to therapy was observed (mean Hb increase 1.6 +/- 0.5 g/dl). In 6 patients, resistance to rhEPO was noted (mean Hb increase 0.5 +/- 0.5 g/dl). The mean gestational age at the start of therapy was 28 weeks of gestation and at the end 32 weeks. The mean duration of a complete therapy was 3.5 weeks (range 2-4.5 weeks). If calculated for body weight, the initial low- rhEPO dose of 160.4 +/- 30.6 U/kg body weight/treatment was increased to 320.9 +/- 61.2 U/kg body weight/treatment in the subsequent phase. CONCLUSION: Response to rhEPO treatment differs widely in anemic pregnant patients with heterozygous hemoglobinopathy. Resistance was observed in anemic pregnant patients with the beta-thalassemia trait originally from the Mediterranean region. PMID- 17119328 TI - Simultaneous hepatic relapse of non-Hodgkin's lymphoma and hepatocellular carcinoma in a patient with hepatitis C virus-related cirrhosis. AB - We report a 66-year-old man with hepatitis C virus (HCV)-related cirrhosis and simultaneous hepatic relapse of non-Hodgkin's lymphoma (NHL) and of hepatocellular carcinoma (HCC). Although the liver is frequently involved by NHL, hepatic colocalization of NHL and HCC is rarely detected by imaging techniques. HCV has been suggested to be lymphotrophic as well as hepatotrophic, and therefore has attracted speculation about a causative role in some cases of lymphoma. The patient had a past history of cutaneous diffuse large B cell lymphoma (DLBCL) in concurrence with HCC 32 months previously. Complete remission (CR) had been maintained for both diseases until February 2004, when ultrasonography and computed tomography (CT) showed multiple liver tumors. Two of these, appearing hyperattenuating in the arterial phase of contrast-enhanced CT, were diagnosed histopathologically as HCC, and treated with radiofrequency ablation. The other tumors, hypoattenuating in the portal phase CT, were diagnosed histopathologically as DLBCL, and treated with cyclophosphamide, tetrahydropyranyl-Adriamycin, vincristine and prednisolone (THP-COP) in combination with rituximab. CR was achieved for both DLBCL and HCC. Given the previously demonstrated immune system tropism and perturbation by HCV, the virus might have contributed to the occurrence of the NHL as well as the HCC. PMID- 17119329 TI - Pseudo-Pelger-Huet in kidney-transplanted patients. AB - Pelger-Huet anomaly is an inherited condition characterized by hyposegmentation of the neutrophil nucleus and excessive chromatin clumping. Acquired Pelger-Huet, also known as pseudo-Pelger-Huet, has been described in several clinical conditions including transplant recipients who received immunosuppressive drugs. The incidence of pseudo-Pelger-Huet in kidney transplant patients, characterized as neutrophil dysplasia, was observed in 9 of 170 patients (5.3%) at the Sao Francisco Hospital de Assis, Belo Horizonte, Brazil. Awareness of possible circulating neutrophil alterations in transplant patients is important for laboratory professionals who should report these findings of cell changes. It should be highlighted that the poor segmentation and the chromatin hypercondensation observed initially in pseudo-Pelger-Huet patients can be suggestive of early-stage neutrophils. Only a combination of laboratory and clinical data will facilitate a better understanding of this anomaly and its correct follow-up and management. PMID- 17119330 TI - Topotecan, Ara-C, cisplatin and prednisolone (Toposhap) for patients with refractory and relapsing lymphomas: Results of a phase II trial. PMID- 17119331 TI - Long-term erythropoietin therapy decreases CC-chemokine levels and intima-media thickness in hemodialyzed patients. AB - BACKGROUND: CC-chemokines are now widely accepted in the recruitment of leukocytes from the blood compartment into tissues, and their role in the progression of atherosclerosis has been documented. Recombinant human erythropoietin (EPO) has become widely used to treat anemic HD patients. However, little is known about the effect of EPO on the plasma CC-chemokine levels and intima-media thickness (IMT) in HD patients. METHODS: Assessment of CC chemokines: monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory proteins (MIP-1alpha, MIP-1beta), regulated upon activation, normal T cell expressed and secreted (RANTES) and IMT were performed in 26 stable HD patients and 15 healthy controls. The patients were divided into 3 groups: group I (n = 8, without EPO), group II (n = 9, EPO at a mean dose of 76 +/- 48 U/kg/week for more than 4 months), and group III (n = 9, EPO at a mean dose of 110.5 +/- 21 U/kg/week for more than 12 months), none of them on iron therapy. RESULTS: MCP-1, MIP-1alpha, MIP-1beta and IMT values were significantly higher, whereas RANTES were significantly lower in HD patients without EPO therapy than those in healthy controls. CC-chemokine levels were found to be significantly lower in patients administered EPO when compared to subjects without EPO. In the patients treated with EPO for more than 12 months IMT values were significantly decreased compared to patients not receiving this hormone. CONCLUSION: These results suggest that long-term EPO therapy decreased CC-chemokine and IMT values in patients undergoing regular HD in the absence of concomitant iron supplementation. PMID- 17119332 TI - Cochlear pharmacokinetics with local inner ear drug delivery using a three dimensional finite-element computer model. AB - HYPOTHESIS: Cochlear fluid pharmacokinetics can be better represented by three dimensional (3D) finite-element simulations of drug dispersal. BACKGROUND: Local drug deliveries to the round window membrane are increasingly being used to treat inner ear disorders. Crucial to the development of safe therapies is knowledge of drug distribution in the inner ear with different delivery methods. Computer simulations allow application protocols and drug delivery systems to be evaluated, and may permit animal studies to be extrapolated to the larger cochlea of the human. METHODS: A finite-element 3D model of the cochlea was constructed based on geometric dimensions of the guinea pig cochlea. Drug propagation along and between compartments was described by passive diffusion. To demonstrate the potential value of the model, methylprednisolone distribution in the cochlea was calculated for two clinically relevant application protocols using pharmacokinetic parameters derived from a prior one-dimensional (1D) model. In addition, a simplified geometry was used to compare results from 3D with 1D simulations. RESULTS: For the simplified geometry, calculated concentration profiles with distance were in excellent agreement between the 1D and the 3D models. Different drug delivery strategies produce very different concentration time courses, peak concentrations and basal-apical concentration gradients of drug. In addition, 3D computations demonstrate the existence of substantial gradients across the scalae in the basal turn. CONCLUSION: The 3D model clearly shows the presence of drug gradients across the basal scalae of guinea pigs, demonstrating the necessity of a 3D approach to predict drug movements across and between scalae with larger cross-sectional areas, such as the human, with accuracy. This is the first model to incorporate the volume of the spiral ligament and to calculate diffusion through this structure. Further development of the 3D model will have to incorporate a more accurate geometry of the entire inner ear and incorporate more of the specific processes that contribute to drug removal from the inner ear fluids. Appropriate computer models may assist in both drug and drug delivery system design and can thus accelerate the development of a rationale-based local drug delivery to the inner ear and its successful establishment in clinical practice. PMID- 17119333 TI - An experimental study of tympanic membrane and manubrium vibrations in rats. AB - Rats are potentially very useful for auditory research because the middle ear structures are easily approachable and because rats are relatively inexpensive. The goal of the present study was to better characterize the mechanics of the rat middle ear by measuring frequency responses at multiple points on the tympanic membrane and manubrium. A laser Doppler vibrometer was used to measure the vibrations. Measurements were made on 7 rats. Tympanic membrane vibrations are presented for 7 different points in the frequency range of 1-10 kHz. The repeatability of the measurements and the interanimal variability at the umbo are also presented. The vibration modes of the tympanic membrane and manubrium were investigated. PMID- 17119334 TI - Vestibular evoked myogenic potentials in newborns. AB - This study presents a novel method for recording vestibular evoked myogenic potential (VEMP) in newborns, used to investigate the maturation of sacculocollic reflex at birth. Twenty full-term newborns aged 2-5 days old were enrolled in this study. During natural sleep, each newborn underwent distortion product otoacoustic emission test, and VEMP test using the head rotation method. For comparison, 20 healthy adults also underwent VEMP test using the same method. Based on adult criteria, 40 newborn ears revealed normal VEMPs in 40%, prolonged VEMPs in 35%, and absent VEMPs in 25%, indicating that great variation exists in the maturation of the sacculocollic reflex at birth. Comparison of VEMP characteristics between healthy newborns and adults revealed nonsignificant difference in the response rate and the latency of n23. However, significant differences existed in the latency of p13, interpeak p13-n23 interval and p13-n23 amplitude between newborns and adults. In conclusion, VEMPs in newborns can be easily recorded by the head rotation method. Prolonged or absent VEMPs in newborns may reflect incomplete maturity of the sacculocollic reflex pathway, especially the myelination. A further large number of newborns receiving MRI scan for other reasons may undergo VEMP test to verify this hypothesis. PMID- 17119335 TI - Vertebral artery dissection after radiation therapy: A case report. PMID- 17119336 TI - Novel mutation in EIF2B gene in a case of adult-onset leukoencephalopathy with vanishing white matter. PMID- 17119337 TI - Balo's encephalitis periaxialis concentrica. AB - In 1928, Balo described a law student with an unusual fatal illness marked by aphasia and a right hemiplegia, with later optic neuritis and normal cerebrospinal fluid. At autopsy, he found a disease of the white matter characterised by foci varying in size from a lentil to that of a pigeon's egg and presenting gray softening and, in part, concentricity, where the medullary sheaths were destroyed and the axis cylinders were intact. He was uncertain whether this was a variant of acute multiple sclerosis or of Schilder's disease. The basis of concentric sclerosis is still unclear though current opinion favours a variant of acute multiple sclerosis. PMID- 17119338 TI - Misoplegia. AB - Amongst the many fascinating abnormal perceptions of parts of the body which may follow strokes and other cerebral lesions is the rare phenomenon of misoplegia. Afflicted patients show dislike, amounting to hatred of the affected part, often accompanied by verbal or physical abuse. Most often the result of a right hemisphere lesion, its precise mechanisms are not well understood. This is a summary of misoplegia and its alleged physiological mechanisms. PMID- 17119339 TI - Urinary retention: a cause of hyponatremia? AB - BACKGROUND: Syndrome of inappropriate antidiuretic hormone secretion (SIADH) is the most common cause of hyponatremia in elderly hospitalized patients; however in many patients the etiology remains unclear even after routine investigations. OBJECTIVE: To report our experience of the association of hyponatremia and urinary retention in elderly hospitalized patients. PATIENTS: Six patients with hyponatremia and urinary retention who were admitted to the geriatric department in our hospital during a four-year period (2001-2004). RESULTS: The mean age of the patients was 85 years. The mean serum sodium level was 120 mEq/l, average volume of retained urine was 933 ml (range 500-1,500 ml). All patients underwent a comprehensive work-up seeking a possible cause for the hyponatremia. The diagnostic work-up was negative. In 5 of the patients the laboratory findings fulfilled the criteria for SIADH. In all patients hyponatremia resolved following urinary catheterization and fluid restriction. All patients made a complete recovery. CONCLUSIONS: Potentially, urinary retention by itself may cause hyponatremia. The possible mechanism for this is SIADH, triggered by bladder distention or pain due to bladder distention. Urinary catheterization may be the key to treatment in these cases of hyponatremia. PMID- 17119340 TI - Human papillomavirus DNA in triple neoplasms of the cervix, perianus and ureter in one woman. AB - BACKGROUND: Multiple malignancies of the urogenital tract have been reported. We investigated the etiological role of human papillomavirus (HPV) in the pathogenesis of urogenital cancers. CASE: We present a case of cervical squamous carcinoma in a 64-year-old woman who subsequently developed Bowen's disease in the perianal skin and transitional cell carcinoma of the ureter. HPV DNA was detected in these tumor specimens using polymerase chain reaction. CONCLUSION: The results strongly suggest that HPV may play a major role in the carcinogenesis of urogenital and perianal tumors. PMID- 17119341 TI - Recent advances in the diagnosis and treatment of pheochromocytoma. AB - Pheochromocytoma (PHEO) is considered to be a rare cause of hypertension. However, if left untreated, PHEOs may lead to fatal hypertensive crises during anesthesia and other stresses. The diagnosis of PHEO is therefore extremely important. A 24-hour blood pressure (BP) pattern per se might be of some diagnostic value due to frequently observed higher BP variability as well as an attenuated night-time BP decrease. So far, germline mutations in five genes have been identified to be responsible for familial PHEOs: the von Hippel-Lindau gene, which causes von Hippel-Lindau syndrome, the RET gene leading to multiple endocrine neoplasia type 2, the neurofibromatosis type 1 gene, which is associated with von Recklinghausen's disease and the genes encoding the B and D subunits of mitochondrial succinate dehydrogenase (SDHB, SDHD), which are associated with familial paragangliomas and PHEOs. Genetic analysis should be offered to those patients with confirmed PHEO who are 50 years old or younger. Plasma-free metanephrines or urinary fractionated metanephrines seem to have higher diagnostic values compared to plasma or urinary catecholamines for the biochemical diagnosis of PHEO. Imaging with (123)I-metaiodobenzylguanidine or (18)F-fluorodopamine PET, if available, are in addition to CT/MRI useful for the detection of multifocal/extra-adrenal forms. Appropriate pharmacologic treatment with subsequent laparoscopic extirpation of PHEO is usually successful in benign forms. There is, however, no convincingly effective mode of treatment in malignant PHEOs. PMID- 17119342 TI - Melatonin and immunomodulation: connections and potential clinical applications. AB - Melatonin is the main hormone secreted by the pineal gland in the human brain. It has a strong impact on the sleep-wake cycle and is considered a general modulator of the human circadian rhythm. Apart from these well-established properties, melatonin possesses immunomodulatory, antioxidative and antiinflammatory properties. The potential ability of this hormone to act synergistically with several cytokines by enhancing their antitumoral activity and dramatically decreasing their adverse effects has placed melatonin among the new and promising agents in cancer immunotherapy. The use of the neurohormone alone or in combination with cytokines and traditional chemotherapeutic drugs is currently under vigorous investigation. Experimental and clinical trials have already depicted some of the immunomodulatory and antitumor effects of melatonin, delineating the need for further research in this field. PMID- 17119343 TI - Modulation of T cell responses in HTLV-1 carriers and in patients with myelopathy associated with HTLV-1. AB - OBJECTIVE: Human T lymphotropic virus-type 1 (HTLV-1) activates the immune system leading to a persistent and exacerbated T-cell response with increased production of IFN-gamma and TNF-alpha. Overproduction of pro-inflammatory cytokines is correlated with the development of HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP), although some HTLV-1 carriers also show high levels of these cytokines. In this study, the ability of regulatory cytokines and cytokine antagonists to inhibit spontaneous IFN-gamma production was investigated. METHOD: IFN-gamma levels were measured by ELISA before and after addition of cytokines or anti-cytokines. RESULTS: Addition of IL-10 significantly reduced spontaneous IFN gamma synthesis in cell cultures from HTLV-1 carriers, while no differences were observed in HAM/TSP patients. There was also a tendency to decreased IFN-gamma levels in cell cultures from HTLV-1 carriers with exogenous addition of TGF-beta. In paired analysis, neutralization of IL-2 significantly decreased IFN-gamma production in HTLV-1 carriers but not in HAM/TSP patients. Neutralization of IL 15 was less effective than neutralization of IL-2 in modulating IFN-gamma production. In HTLV-1 carriers, anti-IL-2 and simultaneous addition of anti-IL-2 and anti-IL-15 decreased IFN-gamma synthesis by 46 and 64%, respectively, whereas in patients with HAM/TSP simultaneous neutralization of both anti-cytokines only decrease IFN-gamma levels by 27%. CONCLUSIONS: Although a large proportion of HTLV-1 carriers produced high levels of pro-inflammatory cytokines similar to those observed in HAM/TSP patients, immune response can be downregulated by cytokines or cytokine antagonists in most HTLV-1 carriers. This modulation can be an important step in the prevention of tissue damage and progression from the HTLV-1 carrier state to HAM/TSP. PMID- 17119344 TI - Role of ammonia and nitric oxide in the decrease in plasma prolactin levels in prehepatic portal hypertensive male rats. AB - OBJECTIVES: Since very little is known about neuroendocrine changes that occur in portal-systemic hepatic encephalopathy, we studied plasma prolactin (PRL) levels and the involvement of hyperammonemia, nitric oxide (NO) and dopaminergic and adrenergic systems in the control of this hormone secretion in a male rat model of prehepatic portal hypertension (PH). METHODS: We conducted in vivo studies to determine plasma ammonia and PRL levels. Dopamine (DA), dihydroxyphenylacetic acid (DOPAC), epinephrine and norepinephrine content in medial basal hypothalamus (MBH) and anterior pituitary (AP) were measured. In addition, NO synthase (NOS) activity and protein expression were evaluated in APs. In in vitro studies, the APs from intact rats were incubated with different doses of ammonia and PRL secretion was determined. In ex vivo studies, the APs from normal and PH rats were incubated in the presence of ammonia and/or a NOS inhibitor, NG-nitro-L arginine-methyl ester (L-NAME) and PRL secretion was determined. RESULTS: PH rats had a significant increase in plasma ammonia levels (p < 0.001) and a decrease in plasma PRL levels (p < 0.05). Neither DA nor DOPAC content or DOPAC/DA ratios were modified in both MBH and APs; however, we observed a significant increase in norepinephrine content in both MBH and AP (p < 0.001 and p < 0.05, respectively) and a significant increase in epinephrine in APs (p < 0.001). Moreover, PH produced an increase in NOS activity (p < 0.01) and NOS protein expression (p < 0.0001) in APs. The ammonia (100 microM) significantly reduced PRL secretion from APs in vitro (p < 0.05). The presence of L-NAME, an inhibitor of NOS, abrogated the inhibitory effect of ammonia on PRL secretion from APs from control and PH rats. CONCLUSIONS: We found that plasma PRL levels were decreased in PH rats probably due to the high ammonia levels. The central noradrenergic system could also mediate this decrease. Also, the increase in NOS activity and/or content in AP induced NO production that directly inhibited PRL secretion from the AP, without the participation of the dopaminergic system. PMID- 17119345 TI - Pathophysiology of acute ischaemic stroke: an analysis of common signalling mechanisms and identification of new molecular targets. AB - Stroke continues to be a major cause of death and disability. The currently available therapies have proven to be highly unsatisfactory (except thrombolysis) and attempts are being made to identify and characterize signalling proteins which could be exploited to design novel therapeutic modalities. The pathophysiology of stroke is a complex process. Delaying interventions from the first hours to days or even weeks following blood vessel occlusion may lead to worsening or impairment of recovery in later stages. The objective of this review is to critically evaluate the major mechanisms underlying stroke pathophysiology, especially the role of cell signalling in excitotoxicity, inflammation, apoptosis, neuroprotection and angiogenesis, and highlight potential novel targets for drug discovery. PMID- 17119346 TI - P16-immunostaining pattern as a predictive marker of lymph node metastasis and recurrence in early uterine cervical cancer. AB - OBJECTIVES: We investigated the relationship between P16-immunostaining patterns and clinicopathological factors in early uterine cervix cancers and assessed whether P16-immunostaining patterns predict the prognosis of the patients with early uterine cervix cancers. METHODS: Twenty-nine early squamous cell carcinoma (SCC) specimens of the uterus were examined using immunohistochemistry for P16 expression. The P16-immunostaining pattern was classified into two groups: the homogeneous type and the heterogeneous type. P16-immunostaining patterns were evaluated in different parts of the carcinoma in situ (CIS): the center of the tumor and the front interface of the infiltrating tumor. RESULTS: All specimens were of the homogeneous type in CIS. The P16-immunostaining pattern was significantly of the heterogeneous type in the front interface of the infiltrating tumor with lymphatic invasion, vascular invasion, lymph node metastasis, and recurrence. Regarding the P16-immunostaining patterns in the front interface of the infiltrating tumor, the patients with the heterogeneous type showed a significantly worse prognosis than the patients with the homogeneous type. CONCLUSIONS: The prognosis of patients with early uterine cervical SCC may be predicted by evaluating the P16-immunostaining pattern in the front interface of the infiltrating tumor. PMID- 17119347 TI - Combination analysis of a whole lymph node by one-step nucleic acid amplification and histology for intraoperative detection of micrometastasis. AB - OBJECTIVE: The aim was to develop a more efficient molecular detection system than histological examination (HE) for lymph node (LN) metastasis. METHODS: Cytokeratin (CK) 19 mRNA copy numbers of 5 colon carcinoma cell lines (Lovo, DLD1, WiDr, Colo201 and Colo320) were calculated and compared by one-step nucleic acid amplification (OSNA) and conventional real-time reverse-transcription polymerase chain reaction (RT-PCR). Then, 91 LN submitted for HE from 6 patients with advanced colorectal adenocarcinoma and 64 LN submitted for frozen diagnosis from 47 patients with different malignancies were examined by OSNA and HE. RESULTS: CK19 mRNA copy numbers of all but Colo320 cells detected by OSNA were within double of those detected by RT-PCR. The least cell count of Lovo cells detected at one reaction (2 microl) by OSNA was calculated as 0.8 cells. Carcinoma metastasis showing either HE+ or OSNA+ was detected in 7.9% of the LN from advanced colorectal adenocarcinomas and in 30.0% of the LN for frozen diagnosis from different malignancies; HE-/OSNA+ metastasis was detected in 4.8 and 4.0%, respectively. OSNA analysis of 1 LN could be completed within 40 min. CONCLUSION: A combined analysis of LN by HE and OSNA could increase the sensitivity for detecting micrometastasis during surgery. PMID- 17119348 TI - A comparative study of neovascularisation in atherosclerotic plaques using CD31, CD105 and TGF beta 1. AB - OBJECTIVES: This study aims to identify plaque neovascularisation using antibodies to CD31, CD105 and TGFbeta1, and to compare their patterns of expression. METHODS: Tissue expression of CD31, CD105 and TGFbeta1 was examined immunohistologically in atherosclerotic plaques from 53 patients who had undergone carotid endarterectomy and in 10 controls. RESULTS: CD31 was observed in a proportion of the microvessels within atheroma. The expression of CD105 was barely visible in normal arteries, but was markedly enhanced in atherosclerotic plaques. The vast majority of the microvessels in atheroma were positive for CD105 with pronounced expression around the periphery of the lipid core. In consecutive sections, microvessels showing negative staining for CD31 were positive for CD105. Although TGFbeta1 was seen in the thickened intima, it was more strongly expressed in well-formed fibrous plaques. Consecutive sections showed that some microvessels were stained by both CD105 and TGFbeta1, but in certain areas microvessels were exclusively CD105 positive. CONCLUSIONS: These observations highlight the distinctive expression patterns of CD31, CD105 and TGFbeta1, suggesting their specific roles in the development of atherosclerotic plaques. CD105 is almost universally expressed in microvessels within the atheroma and is therefore a better vascular marker than CD31 and TGFbeta1for assessing neovascularisation in atherosclerotic plaques. PMID- 17119349 TI - Imbalance of matrix metalloproteinase-1 and tissue inhibitor of metalloproteinases: a novel approach for explaining the parenchymal liquefaction of the septic spleen? AB - OBJECTIVE: The causal pathophysiological mechanisms involved in the parenchymal liquefaction of the septic spleen are still far from clear. The balance between matrix metalloproteinases (MMPs) and their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), is largely responsible for the remodelling of tissues. Deregulation of this balance is a characteristic of extensive tissue degradation in certain chronic inflammatory diseases. METHODS: This study focuses on a search for alterations in the balance between MMP-1 (interstitial collagenase) and TIMP-1 by means of immunostaining, by immunoblotting, and by gel zymography. RESULTS: We found a deregulation of the balance between MMP-1 and TIMP-1 in the septic spleen in favor of the active form of MMP-1. CONCLUSION: Our findings suggest that active MMP-1 is involved in collagenolytic extracellular matrix breakdown in the septic spleen. PMID- 17119350 TI - Bcl-2 phosphorylation has pathological significance in human breast cancer. AB - The anti-apoptotic molecule, Bcl-2, is well known to play an important role in the chemoresistance of breast cancer. We have previously demonstrated that phosphorylation of Fas-associated death domain-containing protein (FADD) at 194 serine through c-jun NH2-terminal kinase (JNK) activation sensitizes breast cancer cells to chemotherapy through accelerating cell cycle arrest at G2/M, and that Bcl-2 phosphorylation downstream of JNK/FADD plays an important role in cell growth suppression by paclitaxel. In this study, the clinicopathological association of phosphorylated Bcl-2 (P-Bcl-2) with estrogen, progesterone, c-erbB 2 receptors, p53 expressions and phosphorylated FADD/JNK (P-FADD/JNK) was analyzed immunohistochemically using 107 human breast cancer specimens. Expression of P-Bcl-2 was found to significantly correlate with lymphatic invasion, lymph node metastasis, but not histological differentiation, tumor grade or vascular and fatty invasion. The positivity of P-Bcl-2 was also significantly correlated to that of P-FADD/JNK. Thus, P-Bcl-2 as well as the P FADD/JNK parameter might be useful markers for cancer progression, independent of the hormone receptor status, in human breast cancers. PMID- 17119351 TI - The future in diagnosis and staging of lung cancer: surgical techniques. AB - Surgical techniques remain central to the diagnosis and staging of lung cancer. Clinical situations which invoke the role of surgery include the diagnosis of solitary pulmonary masses, staging of the mediastinum, restaging of the mediastinum and the assessment of resectability. The techniques available include cervical mediastinoscopy, anterior mediastinotomy, video-assisted thoracoscopy and different procedures for intra-operative mediastinal lymph node assessment including systematic nodal dissection, lobe-specific nodal dissection and sentinel node mapping. The staging of lung cancer is continuously evolving as technological advances combine with clinical advances to better stratify patients into treatment and prognostic categories and alter pre-operative investigation algorithms. Although most of the surgical techniques have been around for many years, it is their application in future which is likely to change. The increasing use of positron emission tomography/computed tomography fusion imaging is raising the proportion of patients being shown to have additional lesions that could contraindicate surgical treatment but which require tissue confirmation to exclude a false-positive examination. Many such lesions are amenable to the expanding techniques available to the interventional endoscopist. The relationship between the surgeon and the endoscopist must become closer to ensure that the appropriate technique is used at each point in the patient's pathway. The future of surgical techniques will be driven by: (1) developments in screening and imaging, with a likelihood that more early stage cancers will present and may be amenable to minimally invasive surgical approaches with the possibility of a role for robotics and nanotechnology; (2) improvements in neoadjuvant therapies which will demand flawless mediastinal staging and restaging; (3) advances in molecular biology which, whilst currently requiring that surgery provide samples of tumour and lymph node tissue to fully characterize the disease, do hold the promise that ever smaller amounts of tissue will be required and that eventually the genetic fingerprint will provide a biological ultrastaging to perhaps supersede anatomical staging. PMID- 17119352 TI - Cardiopulmonary integration: the dark side. PMID- 17119353 TI - Oxidative stress during exercise: further proof that being lean is detrimental for chronic obstructive pulmonary disease patients. PMID- 17119354 TI - Dyspnea: the importance of a psychological approach. PMID- 17119355 TI - Sensory and affective aspects of dyspnea contribute differentially to the Borg scale's measurement of dyspnea. AB - BACKGROUND: Recent research has shown that distinct dimensions in the perception of dyspnea can be differentiated; however, most studies to date have only used a global rating scale for the measurement of this sensation. OBJECTIVES: This study examined the different influence of sensory and affective aspects of perceived dyspnea on the commonly used Borg scale, which measures the global perception of dyspnea. METHODS: Dyspnea was induced in 16 healthy volunteers (mean age 26.2 +/- 6.3 years) by breathing through an inspiratory resistive load (3.57 kPa/l/s) in two experimental conditions (attention and distraction). After each of the two conditions the experienced intensity (i.e., sensory dimension) and unpleasantness (i.e., affective dimension) of dyspnea were rated on separate visual analog scales (VAS), followed by a global rating of dyspnea on the Borg scale. Hierarchical multiple linear regression models were calculated to analyze the predictive validity of VAS ratings of intensity and unpleasantness on the Borg scale ratings. RESULTS: When subjects attended to their breathing, only VAS intensity scores showed a significant influence on Borg scale ratings (p < 0.05). In contrast, only the VAS unpleasantness scores showed a significant influence on Borg scale ratings (p < 0.05) when subjects were distracted. CONCLUSIONS: These findings show that sensory and affective aspects of perceived dyspnea differentially influence the global measure of dyspnea as determined by the Borg scale. A differentiation between these aspects in future studies through the use of separate rating scales could yield more detailed information on the perception and report of dyspnea. PMID- 17119356 TI - Hill walkers' lung. AB - A previously healthy female presented with a 7-week history of dyspnoea on exertion following inhalation of a fluorochemical-based water repellent spray, which was applied to footwear, in the living area that she shared with 8 members of her family while on a hill walking holiday. Clinical examination, serial lung function studies, bronchoalveolar lavage, transbronchial biopsies and high resolution CT thorax confirmed a sub-acute interstitial pneumonitis, which did not resolve until 15 weeks following exposure. None of her family members were affected despite similar exposure. Interstitial pneumonitis due to inhalation of fluorochemical-based water repellent, though rarely described, usually presents in an acute severe form necessitating immediate therapy and resolves in 1-4 weeks. Pulmonary fibrosis can also occur. Sub-acute interstitial pneumonitis following inhalation of fluorochemical-based water repellent spray should always be considered as a cause of unexplained persistent respiratory symptoms in otherwise healthy individuals involved in outdoor pursuits. PMID- 17119357 TI - The pediatric athlete--are we doing the right thing? PMID- 17119358 TI - Managing anterior cruciate ligament deficiency in the skeletally immature individual: a systematic review of the literature. AB - OBJECTIVE: To perform a systematic review of the literature to answer whether early ACL reconstruction for a skeletally immature individuals result in improved outcome compared with nonsurgical treatment or delaying the reconstruction until skeletal maturity in the pediatric athlete. DATA SOURCES: Articles were restricted to the English language, and 6 databases were searched (MEDLINE, CINAHL, EMBASE, ACP Journal Club, Cochrane Central Register of Controlled Trials, and the Cochrane Database of Systematic Reviews). The following keywords were used: anterior cruciate ligament, ACL, child, immature, pediatric, pediatric, and young. SELECTION CRITERIA: All studies examining the ACL reconstruction in skeletally immature individuals, including citations describing complications. Excluded were articles looking at ACL repairs and ACL avulsion injuries. All types of study design, including review articles, were included. Comparative studies were reviewed in detail and analyzed qualitatively. DATA EXTRACTION AND SYNTHESIS: A total of 615 articles were identified. Sixty-six articles met the inclusion and exclusion criteria. There were no articles with levels of evidence better than level III. The majority of the articles represented case series and reviews with expert opinion. RESULTS: There were 7 articles that provided comparisons between surgical and nonsurgical treatment in order to answer the question. CONCLUSIONS: The study designs are inadequate to answer the question of whether early or delayed ACL reconstruction results in the best possible outcome in skeletally immature individuals. Future prospective studies are required to answer the question adequately. A proposed algorithm was described to deal with the question. PMID- 17119359 TI - Articular cartilage repair in the adolescent athlete: is autologous chondrocyte implantation the answer? AB - OBJECTIVE: To determine the evidence base for recommendations regarding autologous chondryocyte implantation in adolescent athletes. MATERIALS AND METHODS: All literature on articular cartilage repair from MEDLINE search dated 1990 to 2006 was reviewed. The majority of articles describe surgical technique and indications. Three techniques for secondary articular cartilage repair have been identified: autologous chondrocyte implantation, autologous osteochondral implants, and marrow stimulation techniques. The initial literature search identified 4 studies that reported the effectiveness and durability of autologous chondrocyte implantation in adults and 2 studies that reported the outcomes of autologous chondrocyte implantation in adolescent athletes. No results of osteochondral implantation or marrow stimulation techniques in adolescent athletes have been published. RESULTS: Acceptable repair rates with all 3 techniques have been reported in adult athletes. Two studies reported high success using autolgous chondrocyte implantation (ACI) in children. CONCLUSIONS: Articular cartilage injury in young athletes remains a difficult problem. The ideal situation is early diagnosis and primary repair, particularly with lesions of the knee, elbow, and ankle. In cases where primary repair is not possible or has been unsuccessful and the lesion is large or symptomatic, secondary repair with either marrow stimulation, microfracture, autologous chondrocyte implantation, or autologous osteochondral grafting may be used. However, at present only the results of ACI repair have been reported for adolescent athletes. PMID- 17119360 TI - The pediatric overhead athlete: what is the real problem? AB - OBJECTIVE: The purposes of this article are to examine shoulder and elbow injuries in pediatric athletes, to evaluate the pathophysiology and pathomechanics that may be associated with the pathoanatomy, and to present suggestions for the prevention of those injuries. DATA SOURCES: This article will review the published sports medicine literature on these topics in baseball and tennis injuries and present a kinetic chain-based perspective on the possible causative factors that are present in the young thrower. RESULTS: The published literature shows that there are multiple intrinsic and extrinsic factors that contribute to the risk of injury in young throwing athletes. These factors appear to develop over time; if not recognized or addressed early, they have undesirable outcomes. CONCLUSIONS: A multitude of factors, including anatomical, biomechanical, and environmental concerns, can contribute to the dysfunction of the shoulder and elbow in young overhead athletes. Understanding the force generating and load-absorbing processes of the body will help clinicians, coaches, and others prevent or limit the deleterious effects of such occurrences. PMID- 17119361 TI - Weight training in youth-growth, maturation, and safety: an evidence-based review. AB - OBJECTIVE: To review the effects of resistance training programs on pre- and early-pubertal youth in the context of response, potential influence on growth and maturation, and occurrence of injury. DESIGN: Evidence-based review. METHODS: Twenty-two reports dealing with experimental resistance training protocols, excluding isometric programs, in pre- and early-pubertal youth, were reviewed in the context of subject characteristics, training protocol, responses, and occurrence of injury. RESULTS: Experimental programs most often used isotonic machines and free weights, 2- and 3-day protocols, and 8- and 12-week durations, with significant improvements in muscular strength during childhood and early adolescence. Strength gains were lost during detraining. Experimental resistance training programs did not influence growth in height and weight of pre- and early adolescent youth, and changes in estimates of body composition were variable and quite small. Only 10 studies systematically monitored injuries, and only three injuries were reported. Estimated injury rates were 0.176, 0.053, and 0.055 per 100 participant-hours in the respective programs. CONCLUSION: Experimental training protocols with weights and resistance machines and with supervision and low instructor/participant ratios are relatively safe and do not negatively impact growth and maturation of pre- and early-pubertal youth. PMID- 17119362 TI - Sports injuries in high school athletes: a review of injury-risk and injury prevention research. AB - OBJECTIVE: The objective of this review is to identify the available research regarding the risk factors and prevention of injuries in high school athletes (ages 14 to 18 years). DATA SOURCES: Relevant manuscripts were identified by searching six electronic databases with a combination of key words and medical subject headings (high school, adolescent, athletic injury, sports injury, risk factors, prevention, and prospective). STUDY SELECTION: Original research that reported prospective data on high school athletes (ages 14 to 18), reported injury and exposure data, and used data collected throughout the entire sport season or school year. DATA EXTRACTION: Twenty-nine studies that identified injury risk factors or injury prevention strategies were reviewed and summarized. Data extracted from the studies included a) sport(s) or injuries studied, b) year of publication, c) lead author, d) description of the subjects, e) sample-size calculation, f) variables studied (baseline demographic or performance variables), g) whether multivariate analyses were used, h) data reported (injury rates, risk ratios, and 95% CI), and i) results. Studies that introduced an intervention were characterized by the same data as well as the type of intervention employed and randomization procedures used. DATA SYNTHESIS: The quality of each injury-risk and injury-prevention study was assessed, and the results were summarized. CONCLUSIONS: The risk factors for injury in several specific sports such as soccer, American football, and basketball have been documented. Other sports are less well represented in the current literature. The risk factors for injuries to the ankle, head, and knee have been identified, to a limited degree. Upper-extremity injury risk factors are less well known. There is a need for high-quality prospective studies to further identify injury risk factors and injury-prevention strategies for high school athletes. PMID- 17119363 TI - Incidence and distribution of pediatric sport-related injuries. AB - OBJECTIVE: To provide a critical review of the available literature on the descriptive epidemiology of pediatric sport-related injuries. DATA SOURCES: MEDLINE (1966 to 2006) and SPORTDiscus (1975 to 2006) were searched to identify potentially relevant articles. A combination of medical subject headings and text words was used (epidemiology, children, adolescents, athletic injuries, sports, injury, and injuries). Additional references from the bibliographies of retrieved articles were also reviewed. STUDY SELECTION: Published research reports on the incidence and distribution of injury in children's and youth sports. Specific emphasis was placed on reviewing original studies, which report incidence rates (rate of injuries per unit athlete time). Forty-nine studies were selected for this review. DATA EXTRACTION: Data summarized include incidence of injury relative to who is affected by injury (sport, participation level, gender, and player position), where injury occurs (anatomical and environmental location), when injury occurs (injury onset and chronometry), and injury outcome (injury type, time loss, clinical outcome, and economic cost). DATA SYNTHESIS: There is little epidemiological data on injuries for some pediatric sports. Many of the studies retrieved were characterized by methodological short-comings and study differences that limit interpretation and comparison of findings across studies. Notwithstanding, the studies reviewed are encouraging and injury patterns that should be studied further with more rigorous study designs to confirm original findings and to probe causes of injury and the effectiveness of preventive measures. CONCLUSIONS: Incidence and severity of injury are high in some child and youth sports. This review will assist in targeting the relevant groups and in designing future research on the epidemiology of pediatric sports injuries. Well designed descriptive and analytical studies are needed to identify the public health impact of pediatric sport injury. PMID- 17119364 TI - Injury prevention in child and adolescent sport: whose responsibility is it? AB - OBJECTIVE: Sport and recreational injuries are a leading cause of morbidity in youth. There is a significant body of literature on risk factors for sport related injuries and a growing body of research supporting the effectiveness of sport-specific prevention strategies in youth. Given the predictability and preventability of injuries in youth sport, the purpose of this article is to develop a model that considers societal responsibility for injury prevention in youth sport, and to discuss the evidence that supports this model. DATA SOURCES/SYNTHESIS: Previously published papers have provided a basis for expert opinion to discuss an approach to examining the shared societal responsibility for implementing countermeasures to reduce the risk of injury to youth during sports. RESULTS: Based on a historical perspective, broad conceptual framework, and specific evidence for prevention strategies in youth sport, the authors have developed and supported a theoretical model that defines a responsibility hierarchy in preventing injuries in youth sport. An argument has been made for a hierarchy of responsibility, with the lowest level of responsibility assigned to the child, and the highest level to those organizations or groups with the potential to effect the most change. The justification for this approach has been discussed in the context of the desirability of passive prevention strategies, the limited evidence for the effectiveness of strategies relying solely on behavior change in children and parents, and the level of perceptual and cognitive development in children that inadequately prepares them to take primary responsibility for their own safety in sport. CONCLUSIONS: The development of effective programs to reduce the burden of sport injury among youth necessitates a scientific approach, the identification of key risk factors for injury, a thorough examination of how factors interact to affect risk, and the identification of potential barriers to the effectiveness of injury-prevention programs. PMID- 17119365 TI - Parental and coach support or pressure on psychosocial outcomes of pediatric athletes in soccer. AB - OBJECTIVE: The purpose of this article was to examine supportive and/or pressuring influences of parents and coaches on young athletes' maladaptive perfectionist tendencies, relationships to friends, and competency perceptions in soccer. Previous research has revealed that parents and coaches may give rise to both enjoyable and stressful sport experiences for the pediatric athlete and that parents and coaches are thus able to influence whether young people decide to quit sport or continue participating. Less is known about the relation of supportive versus pressuring parental and coach behaviors on the quality of athletes' achievement striving, relationships to friends in sport, and their competence perceptions. Such knowledge may help create a better psychological sport experience for pediatric athletes. DATA SOURCES/SYNTHESIS: A questionnaire based cross-sectional field study was carried out among 677 young Norwegian soccer players (aged 10 to 14 years; 504 boys, 173 girls; mean age: boys = 11.9 years, SD = 2.9; girls = 11.2 years, SD = 2.1) taking part in the Norway Cup international youth soccer tournament in 2001. Multivariate analysis of variance (MANOVA) with follow-up canonical correlation was used to examine multivariate relationships between supportive and pressuring behavior and athletes' psychosocial experiences. RESULTS: Joint pressuring behaviors from parents and coaches related positively to maladaptive achievement striving, as indicated by overconcern for mistakes, doubt about one's soccer actions, and lowered perceptions of soccer competence. Mirroring these findings, predominantly supportive coach-created psychological climates were related to a linear pattern of psychological outcomes comprising high-quality friendships, positive competency perceptions, and the absence of specific worries related to achievement striving. CONCLUSIONS: Supportive, mastery-oriented coach influence seems beneficial for constructive psychosocial outcomes in pediatric athletes, and athletes experiencing a joint social pressure to excel from coaches and parents may benefit less psychosocially through sport. PMID- 17119366 TI - Moderate to vigorous exertion and sudden cardiac death in women. PMID- 17119367 TI - Are glucosamine and/or chondroitin sulfate effective for knee osteoarthritis? PMID- 17119368 TI - Predictors of adverse reactions to chiropractic treatment of neck pain. PMID- 17119369 TI - Patellar bracing for patellofemoral pain syndrome. PMID- 17119372 TI - Overview: drug eluting stents. PMID- 17119373 TI - Drug-eluting stents and stent thrombosis: a cause for concern? AB - Drug-eluting stents, most commonly sirolimus-eluting stents and polymer-based paclitaxel-eluting stents, are now widely used during percutaneous coronary interventions, and have largely replaced bare-metal stents to treat a variety of native coronary artery and saphenous vein graft lesions. Stent thrombosis, a complication of both bare-metal and drug-eluting stents, is associated with significant morbidity and mortality including high rates of myocardial infarction and death. Recently, several studies in the literature have raised concern about increased rates of overall stent thrombosis and late stent thrombosis in drug eluting stents in the so-called 'real world' where off-label uses of drug-eluting stents are common. Hypersensitivity reactions to the polymers used in drug eluting stents, delayed endothelialization of the stents, and discontinuation of dual antiplatelet therapy have all been implicated in the pathophysiology of drug eluting stents stent thrombosis. The incidence of total stent thrombosis as well as late stent thrombosis, however, does not seem to be significantly higher in drug-eluting stents than in bare-metal stents. An important risk factor for stent thrombosis in both types of stents appears to be the premature discontinuation of dual antiplatelet therapy, and physicians should educate their patients about the importance of adhering to dual antiplatelet therapy, given the dire clinical consequences of stent thrombosis. PMID- 17119374 TI - Drug-eluting stents for ST[corrected]-elevation acute myocardial infarction: do we need randomized trials? AB - Since their introduction, drug-eluting stents have rapidly altered modern medicine's approach to coronary artery disease. Before the development of drug eluting stents, standard bare-metal stents were plagued by in-stent restenosis, requiring repeat revascularization in as many as 15-20% of patients during the first 6-12 months following implantation [1]. The currently approved drug-eluting stents have dramatically reduced this complication by using a polymer-impregnated coating that elutes either paclitaxel or sirolimus to inhibit smooth muscle proliferation. The pivotal TAXUS-IV [2] and SIRIUS [3] trials compared drug eluting stents with standard bare-metal stents and found rates of target vessel revascularization ranging from 3 to 4.1% in stable coronary artery disease patients - far lower than that had been seen previously with conventional standard bare-metal stents. After their approval in April 2003, drug-eluting stents use in clinical practice expanded rapidly. Within 9 months of their introduction, drug-eluting stents comprised 35% of all stent implantations in the United States [4]. In the last year at our own institution, drug-eluting stents comprised over 85% of all stents implanted. Despite their extensive use, data regarding the efficacy and safety of drug-eluting stents in certain clinical scenarios are limited. To date, the only published data supporting drug-eluting stents in ST[corrected]-elevation acute myocardial infarction come from the retrospective Rapamycin-Eluting Stent Evaluated at Rotterdam Cardiology Hospital registry [5] and the randomized, controlled single high-dose bolus tirofiban and sirolimus-eluting stent vs. abciximab and bare-metal stent in myocardial infarction study [6]. In this chapter, we discuss the theoretical risks and benefits of drug-eluting stents for ST elevation acute myocardial infarction, the available data regarding their use, and the areas in which future studies are needed. PMID- 17119375 TI - From randomized trials to routine clinical practice: an evidence-based approach for the use of drug-eluting stents. AB - The availability of drug-eluting stents has resulted in a paradigm shift in the management of patients with coronary artery disease with a substantial increase in the percentage of patients being revascularized percutaneously rather than surgically. Since its introduction, there has been a tremendous increase in the use of drug-eluting stents with nearly 90% of patients in the US who undergo percutaneous interventions receiving drug-eluting stents. The promising results of several randomized trials that demonstrated a profound reduction in restenosis rates compared with bare-metal stents, underscores the unprecedented enthusiasm among the cardiology community to adopt this new technology swiftly. Data regarding the safety and superiority of drug-eluting stents abound, and it is imperative for the practicing clinician to review and apply them in appropriate clinical settings. In this review, we present general concepts of drug-eluting stents, and attempt to summarize the available data on the approved drug-eluting stents in a variety of patient and lesion subsets. In addition, we share some insights regarding the potential limitations, and issues specific to drug-eluting stents. PMID- 17119376 TI - Drug-eluting stent era: will we improve 5-year outcomes? AB - Five-year outcomes after coronary stenting are determined by restenosis of the original stented lesion during the first year and, later, by disease progression at non-stented segments, owing to either gradual progression of atherosclerosis or instability of vulnerable plaques. Drug-eluting stents have demonstrated potent anti-restenosis benefits in a variety of lesion types and high-risk patients, including complex long lesions and diabetic patients. It is likely that this benefit will translate into improved 5-year outcomes, with reduction in need for repeat revascularization in many patients and possibly reduced incidence of myocardial infarction and death, especially in diabetic patients, in whom the risk for occlusive restenosis and 5-year death and myocardial infarction rates are known to be higher after bare-metal stents. Future studies will determine the role of drug-eluting stents in preventing adverse outcomes owing to later disease progression. Such strategies and proposed clinical trials may include identification and prophylactic stenting of individual vulnerable plaques or coronary segments, and comparisons with coronary artery bypass surgery for complete revascularization success. While the outlook for benefit of drug-eluting stents in 5-year outcomes is hopeful, it is likely that a true improvement in these outcomes will be realized more from a judicious use of drug-eluting stents coupled with other proven aggressive secondary prevention therapies. PMID- 17119377 TI - Increased levels of cardiac troponin-T in outpatients with heart failure and preserved systolic function are related to adverse clinical findings and outcome. AB - BACKGROUND: The implications of increased levels of cardiac troponin T in congestive heart failure with preserved systolic function have been poorly evaluated. We hypothesized that its presence might be related to disease severity and prognosis in this setting. METHODS: Clinical, echocardiographic, 6-min walking test and laboratory data were prospectively obtained in 69 congestive heart failure outpatients with ejection fraction > or = 40%. Serial blood samples were assayed for cardiac troponin T with a third-generation immunoassay and values > or = 0.02 ng/ml were considered abnormal. RESULTS: Abnormal cardiac troponin T levels in at least one sample were found in 27 patients (39%, group 1). These patients were older (71.7 +/- 11 vs. 63 +/- 12.4 years, P = 0.002); more frequently hospitalized during the previous year (63 vs. 26.2%, P = 0.003), had lower systolic blood pressure (129.3 +/- 19.6 vs. 140.4 +/- 23.5 mmHg, P = 0.04), but had similar proportion of ischemic etiology (55.6 vs. 42.9%, P = 0.21) than those with normal cardiac troponin T (group 2). In groups 1 and 2, the functional class was 2.8 +/- 0.8 and 2.1 +/- 0.9 (P = 0.03), and the distance covered in 6 min was 339 +/- 100 and 386 +/- 103 m (P = 0.05), respectively. In groups 1 and 2, the 18-month congestive heart failure hospitalization-free survival was 22 and 87%, respectively (log-rank test P = 0.0003). In a Cox proportional hazard model, functional class III-IV (hazard ratio = 5.21, 95% confidence interval: 1.43-18.96) and myocardial injury (hazard ratio = 5.51, confidence interval: 1.58-19.24) were independently associated with prognosis. CONCLUSION: Increased levels of cardiac troponin T were detected in one out of three congestive heart failure outpatients with preserved systolic function and correlated with clinical measures of disease severity and poor outcome. These findings suggest a link between ongoing myocardial injury and progressive impairment in congestive heart failure despite preserved systolic function. PMID- 17119378 TI - Admission C-reactive protein serum levels and survival in patients with acute myocardial infarction with persistent ST elevation. AB - OBJECTIVE: To evaluate the prognostic value of a single and early determination of high sensitivity C-reactive protein levels at admittance in patients with acute myocardial infarction with persistent ST elevation. PATIENTS AND METHODS: We evaluated high-sensitivity C-reactive protein levels in 247 consecutive acute myocardial infarction with persistent ST elevation patients at admittance. Patients were monitored for the occurrence of major adverse cardiovascular events. RESULTS: Mean follow-up was 26 months. High C-reactive protein levels were principally associated with age > or = 65 years (P=0.01), diabetes (P=0.03) and reduced left ventricle ejection fraction (P=0.048). We observed a significant C-reactive protein level difference between the major adverse cardiovascular event-free group and the major adverse cardiovascular event group (28.2+/-21.9 vs. 47.7+/-31.9 mg/l, P=0.03), between deceased patients group (vs. 81.5+/-51.8 mg/l, P<0.001) and early deaths (vs. 129.5+/-71.9 mg/l, P<0.001). Kaplan-Meier plots for survival and major adverse cardiovascular event occurrence showed a significant separation (P=0.01 and 0.002 by log-rank test, respectively) between high and low C-reactive protein level groups. C-reactive protein levels were independent risk predictors of major adverse cardiovascular events (odds ratio 2.931, 95% confidence interval 1.512-5.893; P=0.046) and death (odds ratio 5.068, 95% confidence interval 2.056-20.195; P=0.04). Patients with high C-reactive protein levels and age > or = 65 years were at highest risk for major adverse cardiovascular event occurrence (odds ratio 5.658, 95% confidence interval 2.898 6.249; P=0.022) and death (odds ratio 8.120, 95% confidence interval 5.656 22.729; P=0.03). CONCLUSIONS: High C-reactive protein levels identify patients with a worse prognosis after acute myocardial infarction with persistent ST elevation. The evaluation of C-reactive protein and age may provide a tool to select high-risk patients. PMID- 17119379 TI - Role of interleukin-17 and interleukin-17-induced cytokines interleukin-6 and interleukin-8 in unstable coronary artery disease. AB - BACKGROUND: Atherosclerosis is a chronic inflammatory disease and interleukins are considered to play a key role in the chronic vascular inflammatory response that is typical of atherosclerosis. The serum levels of several of these cytokines have been found to positively correlate with coronary arterial disease and its sequelae. AIM: The aim of our study was to evaluate the levels of a comparatively new cytokine IL-17, in patients with stable and unstable coronary artery disease in order to assess whether unstable coronary artery disease patients had higher IL-17 levels. MATERIALS AND METHODS: We analyzed the concentrations of IL-17, IL-6, IL-8 and IL-10 using enzyme-linked immunosorbent assay and heat-sensitive C-reactive protein using latex particle-enhanced immunoturbidimetry in 58 consecutive unselected patients divided into three groups: stable angina (n=14), unstable angina (n=24) and acute myocardial infarction (n=20). We further compared them with 20 healthy controls. These 58 patients were also angiographically studied and divided into two groups: simple lesion (n=22) and complex lesion (n=36), on the basis of the coronary plaque morphology. RESULTS: Our results show increased concentrations of the proinflammatory cytokines IL-17, IL-6, IL-8 and heat-sensitive C-reactive protein, and decreased concentration of IL-10 in plasma of unstable angina and acute myocardial infarction patients. Plasma concentration of IL-17 was also positively correlated with plasma concentrations of IL-6 and heat-sensitive C reactive protein. Our findings further showed that IL-17 values were higher in patients having angiographically visible complex types of lesions but no difference was observed between complex and simple lesion morphology patients. CONCLUSION: In conclusion, these findings point towards a role of inflammation in the form of increased activity of IL-17, IL-6 and IL-8 in patients of unstable angina and acute myocardial infarction and thus suggest that IL-17-driven inflammation may play a role in the promotion of clinical instability in patients with coronary artery disease. PMID- 17119380 TI - Diurnal variation of the P-wave dispersion in chronic ischemic heart diseases. AB - OBJECTIVE: Electrocardiographic indices like maximum P-wave duration (P(max)) and P-wave dispersion (PD) can be used to detect patients with atrial conduction disorders, myocardial ischemia and those at risk for atrial fibrillation. Considering the diurnal variation of ischemia in patients with significant coronary lesions, this study was designed to investigate the diurnal variation of eventual atrial conduction abnormalities. METHODS: Forty-eight patients (31 male) with typical angina were grouped according to coronary angiography results as group 1 - 70% or more luminal reduction in at least one of the coronary arteries (n=28), and group 2 - normal coronary arteries (n=20). The difference between the P(max) and minimum P-wave durations (P(min)) is designated as PD. The diurnal P(max), P(min) and PD values were compared between and within the groups. RESULTS: The morning P(max) value of group 1 was significantly higher than the value of group 2 (112+/-1 vs. 102+/-1 ms, P<0.001). The morning PD of group 1 was significantly higher than that of group 2 (54+/-9 vs. 48+/-1 ms, P<0.05). The morning P(max) of group 1 (112+/-1 ms) was significantly higher than its afternoon (102+/-9 ms) and night (102+/-1 ms) values (P<0.001). The morning PD of group 1 (54+/-9 ms) was higher than the afternoon (40+/-10 ms) and night (43+/-9 ms) PD (P<0.001). No significant difference was observed between the P(max), P(min) and PD values in group 2 (P>0.05). CONCLUSION: This study demonstrated that coronary heart disease patients have higher morning P(max) and PD values that may be important regarding prediction of timing and treatment of atrial conduction disorders in myocardial ischemia. PMID- 17119381 TI - Bibliography of current world literature. PMID- 17119382 TI - A population-based study of the frequency of corticosteroid resistance and dependence in pediatric patients with Crohn's disease and ulcerative colitis. AB - BACKGROUND: The goal of this study was to examine the 1-year outcome after the first course of systemic corticosteroids in an inception cohort of pediatric patients with inflammatory bowel disease. METHODS: All Olmsted County (Minnesota) residents diagnosed with Crohn's disease (n = 50) or ulcerative colitis (n = 36) before 19 years of age from 1940 to 2001 were identified. Outcomes at 30 days and 1 year after the initial course of corticosteroids were recorded. RESULTS: Twenty six patients with Crohn's disease (65%) and 14 with ulcerative colitis (44%) were treated with corticosteroids before age 19. Thirty-day outcomes for corticosteroid-treated Crohn's disease were complete remission in 16 (62%), partial remission in 7 (27%), and no response in 3 (12%), with 2 of these patients requiring surgery. Thirty-day outcomes for treated ulcerative colitis were complete remission in 7 (50%), partial remission in 4 (29%), and no response in 3 (21%). One-year outcomes for Crohn's disease were prolonged response in 11 (42%) and corticosteroid dependence in 8 (31%), whereas 7 (27%) were postsurgical. One-year outcomes for ulcerative colitis were prolonged response in 8 (57%) and corticosteroid dependence in 2 (14%), whereas 4 (29%) were postsurgical. CONCLUSIONS: Most pediatric patients with inflammatory bowel disease initially responded to corticosteroids. However, after 1 year, 58% of pediatric patients with Crohn's disease and 43% of pediatric patients with ulcerative colitis either were steroid dependent or required surgery. This finding emphasizes the need for early steroid-sparing medications in pediatric inflammatory bowel disease. PMID- 17119383 TI - Impaired intestinal iron absorption in Crohn's disease correlates with disease activity and markers of inflammation. AB - BACKGROUND: Anemia in patients with Crohn's disease (CD) is a common problem of multifactorial origin, including blood loss, malabsorption of iron, and anemia of inflammation. Anemia of inflammation is caused by the effects of inflammatory cytokines [predominantly interleukin-6 (IL-6)] on iron transport in enterocytes and macrophages. We sought to elucidate alterations in iron absorption in pediatric patients with active and inactive CD. METHODS: Nineteen subjects with CD (8 female, 11 male patients) were recruited between April 2003 and June 2004. After an overnight fast, serum iron and hemoglobin levels, serum markers of inflammation [IL-6, C-reactive protein (CRP), and erythrocyte sedimentation rate], and a urine sample for hepcidin assay were obtained at 8 am. Ferrous sulfate (1 mg/kg) was administered orally, followed by determination of serum iron concentrations hourly for 4 hours after the ingestion of iron. An area under the curve for iron absorption was calculated for each patient data set. RESULTS: There was a strong inverse correlation between the area under the curve and IL-6 (P = 0.002) and area under the curve and CRP levels (P = 0.04). Similarly, the difference between baseline and 2-hour serum iron level (Delta[Fe]2hr) correlated with IL-6 (P = 0.008) and CRP (P = 0.045). When cutoff values for IL-6 (>5 pg/mL) and CRP (>1.0 mg/dL) were used, urine hepcidin levels also positively correlated with IL-6 and CRP levels (P = 0.003 and 0.007, respectively). CONCLUSIONS: Subjects with active CD have impaired oral iron absorption and elevated IL-6 levels compared with subjects with inactive disease. These findings suggest that oral iron may be of limited benefit to these patients. Future study is needed to define the molecular basis for impaired iron absorption. PMID- 17119384 TI - A practical perspective on ulcerative colitis: patients' needs from aminosalicylate therapies. AB - A large, Internet-based survey of a random sample of members of the Crohn's and Colitis Foundation of America was undertaken to gain knowledge and understanding of patients' experiences with ulcerative colitis and first-line therapies. From 49,410 invitations to participate, 1,595 usable responses were received from patients with ulcerative colitis. Patients were prescribed a range of aminosalicylates for their ulcerative colitis. Treatments with the highest proportion of satisfied patients were associated with highest remission rates. Forty-three percent of patients considered their disease to be in remission; however, 74% reported disease relapse during the previous 12 months. Over 60% of patients reported that they were noncompliant with prescribed aminosalicylate dosing schedules, with reasons attributed to frequency of dosing, the number of pills, and the inconvenience of the medication. Many respondents reported that they had made significant lifestyle changes because of their ulcerative colitis, including spending more time at home (46%) and participating in fewer social activities (37%). When asked to describe their ideal treatment, patients considered high efficacy (97%), lack of side effects (74%), nonparenteral dosing (46%), nonrectal dosing (36%), low cost (23%), fewer pills (23%), and less frequent dosing (23%) as "very important." This study demonstrates that continuous symptomatic remission is central to patient satisfaction and that patients find currently available aminosalicylates to be inconvenient. Patients' ideal therapy would be an effective, oral formulation with fewer tablets, less frequent dosing, and minimal side effects. Development of such a therapy would, therefore, potentially improve both patient compliance and overall treatment success. PMID- 17119385 TI - Predictive value of the CARD15 variant 1007fs for the diagnosis of intestinal stenoses and the need for surgery in Crohn's disease in clinical practice: results of a prospective study. AB - BACKGROUND AND AIM: The diagnostic and therapeutic relevance of CARD15 genotyping in Crohn's disease (CD) for daily clinical practice has not been investigated so far. We therefore analyzed whether CARD15 variants are independent predictive factors for small bowel stenosis in CD evaluated by magnetic resonance enteroclysis (MRE). On the basis of these findings, the potential implications for patient management were investigated. METHODS: Eighty CD patients with clinical symptoms suggestive of small bowel stenosis were included. All patients were genotyped for the CARD15 variants c.2104C > T (p.R702W), c.2722G > C (p.G908R), and c.3019_3020insC (p.Leu1007fsX1008) and examined by MRE of the small bowel. RESULTS: CARD15 variants were found in 40 (50%) patients. MRE identified 31 (38%) patients with small bowel stenoses. Twenty-five of the 40 (62%) patients with at least one CARD15 variant were diagnosed of intestinal stenosis by MRE (odds ratio [OR] = 9.44; confidence interval [CI] 3.21-27.77; P = 0.00028, Bonferroni corrected). Particularly, the presence of the 1007fs variant was associated with an increased risk of an intestinal stenosis (OR = 12.00, CI 3.47-41.54, P = 0.00042, Bonferroni corrected). Twenty-one of 31 (68%) patients with stenoses required surgical intervention, with 13 of these 21 (62%) patients carrying the 1007fs variant. CONCLUSION: In the largest prospective study analyzing the diagnostic value of CARD15 variants in CD patients performed so far, we identified the 1007fs variant as strong predictor for intestinal stenoses with need for surgery in CD patients. Genotyping could therefore be an important diagnostic tool in clinical practice for identifying high-risk patients with specific diagnostic and therapeutic needs. Moreover, MRE is an excellent technique for diagnosing small bowel stenoses. PMID- 17119386 TI - Antibodies to I2 predict clinical response to fecal diversion in Crohn's disease. AB - OBJECTIVES: Fecal diversion is occasionally indicated in patients with advanced perianal or colorectal Crohn's disease (CD). Because CD may result from an aberrant immunologic response to bacteria within the gut lumen, fecal diversion should be effective in managing these complications. However, not all patients achieve a clinical response after fecal diversion. CD patients can be characterized by their antibody responses against Pseudomonas fluorescens (I2), E.coli outer membrane porin C (OmpC), oligomannan (anti-Saccharomyces cerevisiae antibodies [ASCA]), and antinuclear antigens (perinuclear antineutrophil cytoplasmic antibodies [pANCA]). This study examines the association between clinical features and seroreactivity to these microbial and auto-antigens in predicting a clinical response to fecal diversion. METHODS: Twenty-seven consecutive CD patients undergoing fecal diversion were included. Sera were drawn and tested for anti-I2, anti-OmpC, ASCA, and pANCA in a blinded fashion. Response was assessed using clinical parameters. RESULTS: Seventeen (63%) patients underwent fecal diversion for medically resistant proctocolitis and 10 (37%) for severe perianal disease. Median follow-up was 41 months. Seventeen (63%) patients achieved a clinical response. No preoperative clinical or surgical factor predicted response to diversion. Clinical response after fecal diversion was seen in 15 of 16 (94%) patients who were I2 positive compared with only 2 of 11 (18%) patients who were I2 negative (P = 0.0001). Seroreactivity to OmpC, ASCA, or pANCA was not associated with a clinical response to diversion. CONCLUSION: Expression of I2 antibodies against a bacterial antigen of Pseudomonas fluorescens was highly associated with clinical response to fecal diversion in CD patients. PMID- 17119387 TI - Treatment of ulcerative colitis refractory to steroid therapy by oral microemulsion cyclosporine (Neoral). AB - BACKGROUND: Intravenous cyclosporine is active in 60% to 80% of patients with ulcerative colitis (UC) who failed to respond to intravenous corticosteroids. Several studies have suggested that cyclosporine in microemulsion form (Neoral) has some efficacy in this setting, but the optimal dose, blood level, time to response, and remission need to be better defined. The aim of this study was to evaluate the response to Neoral and its toxicity in active corticosteroid refractory UC. METHODS: Between March 2002 and August 2005, 20 courses of Neoral [initial dose, 2.3 mg/kg (range, 1.8 to 2.8 mg/kg) every 12 hours] were prescribed in 19 consecutive patients for a UC attack that did not respond to intravenous methylprednisolone. All patients received prophylaxis against Pneumocystis carinii. RESULTS: Response was obtained in 17 of 20 attacks (85%) after 3.5 days (range, 1 to 7). Remission was obtained in 15 of 20 attacks (75%) after 13 days (range, 2 to 30 days). Four responders relapsed and underwent colectomy 21 to 900 days after the start of Neoral. Overall, 14 of 19 patients (74%) were colectomy free after a median follow-up of 8 months (range, 1 to 41 months). Cyclosporine blood levels were measured at fasting (C0) and 2 hours after Neoral administration (C2) in a subgroup of 10 responders. The results were 103 ng/mL (range, 32 to 240 ng/mL) for C0 and 761 ng/mL (183 to 1390 ng/mL) for C2. One severe bedridden patient with neonatal encephalopathy died. Main side effects observed were mild transient renal impairment (n = 2), hypertension (n = 1), cytomegalovirus infection (n = 2), and esophageal candidiasis (n = 1). CONCLUSIONS: In active corticosteroid-refractory UC, Neoral seems to have the same efficacy and toxicity as the intravenous form. Trough target cyclosporine blood levels should not exceed 100 ng/mL for C0 and 700 ng/mL for C2. PMID- 17119388 TI - Abnormal microbiota composition in the ileocolonic mucosa of Crohn's disease patients as revealed by polymerase chain reaction-denaturing gradient gel electrophoresis. AB - BACKGROUND: Bacteria might play a role in the pathogenesis of Crohn's disease (CD), and patients harbor a different type and density of gut microbiota compared with normal healthy subjects. Thus, the aim of this study was to compare the microbiota adhered to the mucosa of CD patients with that of healthy subjects. METHODS: Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR DGGE) of 16S rRNA gene fragments was used to identify the dominant bacterial species present in fresh biopsy samples obtained from the mucosa of 15 healthy and 19 CD subjects. Two patients suffering from ulcerative colitis and 1 suffering from ischemic colitis also were included. RESULTS: Individuals were clustered in 2 groups according to their molecular fingerprint, which differentiated the majority of CD specimens (88.2%) from the majority of healthy/ulcerative colitis/ischemic colitis specimens (82.3%). In addition, the patient-to-patient variability in microbiota was greater within the CD cluster than in the healthy/ulcerative colitis cluster (P = 0.000). One hundred forty-one sequences were obtained from the PCR-DGGE bands that were grouped into 58 different phylotypes, 8 of which were novel. BLAST analysis revealed that 74.5% of the sequences were similar to those of bacteria that have never been cultivated. In CD samples, prevalence values for Clostridium spp Ruminococcus torques and Escherichia coli were significantly higher, whereas Faecalibacterium was more frequently found in healthy specimens. Opportunistic pathogenic gamma proteobacteria were found occasionally, only in CD mucosal microbiota. CONCLUSIONS: Microbiota attached to the ileocolonic mucosa of CD patients is distinguishable from that of healthy subjects. We postulate that individuals who are predisposed to CD are less able to regulate the microbial makeup of their intestines, which leads to an unstable microbial population. PMID- 17119389 TI - Familial occurrence of inflammatory bowel disease in Korea. AB - BACKGROUND: Little information is available about the familial aggregation of inflammatory bowel disease (IBD) in Asian populations. We therefore determined the risk of familial aggregation of IBD among first-degree relatives of patients with ulcerative colitis (UC) or Crohn's disease (CD) in an ethnically distinct Korean population. METHODS: Familial aggregation of IBD was evaluated in terms of family history, prevalence, lifetime risk, and population relative risk in first degree relatives of 1440 unrelated patients with UC (n = 1043) or CD (n = 397). RESULTS: A positive first-degree family history of IBD was observed in 27 probands (1.88%): 21 of 1043 (2.01%) with UC and 6 of 397 (1.51%) with CD. The crude prevalence of IBD in first-degree relatives of probands with IBD was 0.31%. The lifetime risk of IBD was 0.54% in all first-degree relatives of IBD probands, 0.52% in UC probands, and 0.67% in CD probands, with overall lifetime relative risks of 0.12% in parents, 0.79% in siblings, and 1.43% in offspring. The age- and sex-adjusted population relative risk of IBD was 13.8 in first-degree relatives of probands with IBD. CONCLUSIONS: Although a positive family history, prevalence, and lifetime risk of IBD among first-degree relatives of Korean IBD patients are much lower than among relatives of Western patients, the population relative risk in first-degree relatives is about equal in Koreans and Westerners. This finding indicates that a positive family history is an important risk factor for IBD in Koreans and in Westerners. PMID- 17119390 TI - Plant sterol guggulsterone inhibits nuclear factor-kappaB signaling in intestinal epithelial cells by blocking IkappaB kinase and ameliorates acute murine colitis. AB - BACKGROUND/AIMS: The plant sterol guggulsterone has been shown to have anti inflammatory properties. It remains unknown, however, whether guggulsterone is effective for the treatment of inflammatory bowel disease (IBD). Therefore, we investigated anti-inflammatory effects of guggulsterone on intestinal epithelial cells (IEC) and on experimental murine colitis models and elucidated its molecular mechanisms. METHODS: Human Caco-2 cells and rat non-transformed IEC-18 cells were stimulated with interleukin (IL)-1beta or lipopolysaccharide (LPS) with or without guggulsterone. The effects of guggulsterone on nuclear factor (NF)-kappaB signaling in IEC were examined by intercellular adhesion molecule (ICAM)-1 real-time reverse-transcription polymerase chain reaction, NF-kappaB transcriptional activity assay, Western blotting for IkappaB phosphorylation/degradation, electrophoretic mobility shift assay, and in vitro IkappaB kinase (IKK) assay. For in vivo study, dextran sulfate sodium (DSS) treated mice were fed with or without guggulsterone. Colitis was quantified by disease activity index and evaluation of macroscopic and microscopic findings. Phosphorylation of IkappaB and IKK in colon mucosa was assessed by Western blotting and immunohistochemistry. RESULTS: Guggulsterone significantly inhibited LPS- or IL-1beta-induced ICAM-1 gene expression, NF-kappaB transcriptional activity, IkappaB phosphorylation/degradation, and NF-kappaB DNA binding activity in IEC. Moreover, guggulsterone strongly blocked IKK activity. Administration of guggulsterone significantly reduced the severity of DSS-induced murine colitis as assessed by clinical disease activity score, colon length, and histology. Furthermore, tissue upregulation of IkappaB and IKK phosphorylation induced by DSS was attenuated in guggulsterone-treated mice. CONCLUSION: Guggulsterone blocks NF-kappaB signaling pathway by targeting IKK complex in IEC and attenuates DSS-induced acute murine colitis, which suggests that guggulsterone could be an attractive therapeutic option in the treatment of IBD. PMID- 17119391 TI - Report on the vitamin D status of adult and pediatric patients with inflammatory bowel disease and its significance for bone health and disease. AB - Vitamin D is a hormone responsible for calcium homeostasis and essential for bone mineralization throughout the lifespan. Recent studies revealed a high prevalence of hypovitaminosis D among healthy adults and children, especially in the northern hemisphere, and a link between this condition and suboptimal bone health. Moreover, maintenance of what are today considered optimal vitamin D stores has not been achieved throughout the year with currently recommended daily intake for vitamin D. The prevalence of hypovitaminosis D is even higher among adults with inflammatory bowel disease (IBD), a situation that may be caused by malabsorption and gastrointestinal losses through an inflamed intestine, among other factors. In children with IBD, existing reports of vitamin D status are scarce. The relationship between vitamin D status and bone health, although well established in healthy adults and children, has been controversial among adults and children with IBD, and the reasons for this have not been investigated to date. Studies in animal models of colitis and in vitro human studies support a role of vitamin D in the regulation of the immune system of the gut and the potential of vitamin D and its derivatives as therapeutic adjuncts in the treatment of IBD. This role of vitamin D has not been investigated with translational studies to date. Currently, there are no guidelines for monitoring vitamin D status, treating hypovitaminosis D, and maintaining optimal vitamin D stores in patients with IBD. These tasks may prove particularly difficult because of malabsorption and gastrointestinal losses that are associated with IBD. PMID- 17119392 TI - Psychotherapy with chronic inflammatory bowel disease patients: a review. AB - Two distinct factors have lead in the past to the development of several psychotherapeutic treatments for patients with inflammatory bowel diseases (IBD). First, clinicians and researchers believe that psychologic and somatic factors in chronic IBD, Crohn's disease, and ulcerative colitis) are connected. In addition, IBD reduces the health-related quality of life for these patients. The purpose of the psychotherapies is to influence the somatic course of the disease, the psychological state of the patients, or the patients' health related quality of life. This report evaluates the existing studies with regard to the effectiveness of psychotherapy IBD patients received in addition to medical treatment. We have identified 10 psychotherapy studies and 4 additional studies on self management and patient education on this topic. The studies significantly differ from each other in regard to psychotherapeutic methods, inclusion criteria, and outcome assessments. The results so far lead to the conclusion that psychotherapy does not have an impact on the course of the disease but, in some cases, positively influences the patient's psychologic state (such as depression, anxiety, and health related quality of life or coping with the disease). Thus, psychotherapy cannot, in general, be recommended for all patients with chronic IBD. Patients, however, that display a tendency toward psychologic problems, especially as it pertains to their illness, might profit from it. PMID- 17119393 TI - Crohn's disease: the cost of comfortable inaction. PMID- 17119394 TI - Crohn's disease: is it becoming more common? PMID- 17119395 TI - Crohn's disease associated with severe autoimmune thrombocytopenic purpura. PMID- 17119396 TI - Is intestinal permeability test useful for asymptomatic Crohn's disease? PMID- 17119397 TI - Absence of Mycobacterium avium subspecies paratuberculosis in Crohn's disease patients. PMID- 17119398 TI - Giant nevi: a review. AB - Congenital nevi are benign neoplasms that are present at birth and composed of nevomelanocytes. Approximately 1-3% of all newborns have congenital pigmented nevi, and the number of nevi increases with age, peaking by late adolescence to 20-40 nevi in an individual. Giant congenital nevi are often defined as nevi that are greater than 20 cm in diameter in an adult, or nevi that occupy 2% or more of the body surface area. Histologically, nevi are transformed melanocytes, which are normally highly dendritic cells interspersed among basal keratinocytes. The genetic basis of these lesions is not known. Findings of a culture of melanocytes from such a lesion from a showed chromosome rearrangements involving 1p,12q, and 19p. The giant nevi might be associated to several diseases: neurocutaneous melanosis, diffuse lipomatosis, structural brain malformations, hypertrophy of skull bones, limb atrophy, skeletal asymmetry involving both soft tissue hyper and hypoplasia, von Recklinghausen's disease and vitiligo. The risk of malignant change in giant nevi is probably the most contentious issue in its management. The consensus is that lesions are pre-malignant, but the purported incidence of malignancy varies wildly from 0-42%. Surgical excision remains the mainstay of treatment for large congenital melanocytic nevi, and most giant nevi are managed by staged excision and resurfacing with skin grafts or tissue expanders and flaps. PMID- 17119399 TI - The grand nursery--are we getting there in an effort to get out. PMID- 17119400 TI - Recent advances made in wartime medical care. PMID- 17119401 TI - A new method for assessment of craniofacial malformations. AB - Many assessments of craniofacial malformations are generally undertaken to assist in surgical intervention including physical examination, cephalometric radiographs in anteroposterior and lateral views, stereolithographic models, and anthropometric measurements integrated with three-dimensional computed tomography (3-D CT) reconstructions to quantify skeletal deformities. In the present report, the use of 3-D Malformation Analysis, a three-dimensional methodology for planning craniofacial operative procedures, is presented. In addition to cephalometric and anthropometric databases, the measurements from 3-D surface reconstructions from CT were used intraoperatively to establish the correct position of skeletal segments. PMID- 17119402 TI - Orbital fractures: a new classification and staging of 190 patients. AB - The orbit is located in the middle third of the face, composed of several bones and surrounded by complex anatomic structures so that orbital fractures (OF) often involve other parts of the face. A staging system for classifying OF is of paramount importance in order to exchange information between trauma centers. Several classifications have been proposed for describing OF but they have not a single method applicable to the whole orbit. Here, a classification for OF that can be summarized with four abbreviations is proposed. Four letters define the localization (F = frontal, N = nasal, M = maxillary and Z = zygomatic bone fracture), two acronyms describe fragment shift (in = blow-in or out = blow-out), four numbers define ocular movement impairment (1 = superior, 2 = internal, 3 = inferior, and 4 = external extrinsic muscular deficit) and two acronyms describe eye position (EX = exophthalmos and ENO = enophthalmos). To evaluate the suitability of the proposed classification a retrospective study on a series of 190 OFs is performed. Age, gender, new stage, clinical diagnosis at admission, type of surgery, and need for graft for orbital reconstruction are considered. A good correlation between the proposed classification and the studied variables is detected. In conclusion, the proposed classification is a simply and precise method to stage OF. It can summarize OF and be used in the daily practice. However, it is our belief that a multi-center study should be performed before the effectiveness of the proposed classification can be clearly stated. PMID- 17119403 TI - Short- and long-term effect of surgically assisted maxillary expansion on nasal airway size. AB - Transverse maxillary deficiency (TMD) may reduce nasal dimensions and lead to oral breathing. The objective of the present study was to investigate the short- and long-term effects of surgically assisted maxillary expansion on nasal airway size of patients with TMD. Eleven subjects with TMD and skeletal maturity were submitted to posterior rhinomanometry to determine the minimum nasal cross sectional area (CSA) before and 3, 6, and 12 months after surgery. Subjects were also investigated concerning their habitual diurnal and nocturnal breathing mode (oral, nasal, and oronasal). A statistically significant increase in mean CSA was observed in the early postoperative period (3 months), followed by a decrease to the preoperative levels in the subsequent periods (6 and 12 months). No variations were observed in the breathing mode for 63.6% of the subjects. The results showed that, in the short-term, maxillary expansion most frequently produced an increase in nasal patency. However, it was observed that the effect did not persist over time in most subjects. PMID- 17119404 TI - Le Fort III midfacial distraction using an internal distraction device for syndromic craniosynostosis: device selection, problems, indications, and a proposal for use of a parallel bar for device-setting. AB - Le Fort III midfacial distraction using internal and external devices is a well accepted procedure for the midfacial retrusion of craniosynostosis syndrome patients. The authors described 20 consecutive series of Le Fort III midfacial distraction using internal distraction devices. Two types of devices were utilized. One type was a zygoma-skull device (the anterior part of the device is attached to the zygoma, and the posterior part is attached to the skull), which was used in six cases. The other was a zygoma-zygoma device (the anterior and posterior parts of the device are attached to the osteotomized zygoma, respectively), which was used in 14 cases. Subject ages ranged 3-32 years. A 14 20-mm distraction length was obtained by 1 mm/day. Satisfactory distraction of the midface was obtained in 17/20 cases. In 3/6 cases in which a zygoma-skull device was used, an unsatisfactory result was obtained. In these three cases, a fracture of the zygomatico-maxillary suture was encountered, resulting in the Le Fort III portion being left behind. In all 14 cases in which a zygoma-zygoma device was used, a satisfactory result was obtained. During the distraction period, the connection of the distraction device was dislodged, resulting in re connection in three cases. Slight asymmetry was noticed in two cases without any need for management. In order to obtain parallel setting of the bilateral distraction devices, a newly developed parallel bar was used and demonstrated to be effective. PMID- 17119405 TI - A retrospective analysis of 101 zygomatico-orbital fractures. AB - A 5-year review of 101 cases of zygomatico-orbital fractures is presented. The epidemiology, fracture patterns, treatment modalities, and complications were evaluated in this retrospective study. A majority of fractures were sustained by males and resulted from trauma inflicted during altercations and traffic accidents. The most common fracture pattern was tripod fracture and the most common associated facial fractures were mandibular fractures. Open reduction and rigid fixation was the most frequently employed treatment modality. Depending on the stability of reduced zygoma, one, two and three-point fixations were applied. Orbital floor exploration was performed in 41 cases. Ten out of 16 orbital floor bone defects required reconstruction. In these cases orbital floor was reconstructed with 1.5-mm porous polyethylene implant. Although we encountered a few complications related to the incisions for open reduction, the rate of complication in which correction was difficult (e.g. facial asymmetry) was lower with this approach when compared with the literature. PMID- 17119406 TI - Various applications of chondrocutaneous composite grafts in secondary cleft lip nose patients. AB - Composite chondrocutaneous grafts were applied to 12 patients in various forms to repair the columellar deficit, to form the nasal tubercle and nostril sill in cleft lip nose patients. Cleft lip-nose deformity patients with alar cartilage hypoplasia, obtuse angulation of the medial and lateral crura and the resulting plica vestibularis, internal nasal valve problems associated with the weakness of upper lateral cartilages are included in this study and composite conchal cartilage grafts are utilized to achieve a symmetrical and functional result. PMID- 17119407 TI - Psychiatric analysis of suicide attempt subjects due to maxillofacial gunshot. AB - The studies of maxillofacial gunshot injuries mainly focused on evaluating the surgical interventions and physical outcomes of the procedures. In this study we aimed to analyze the pre- and post-injury psychiatric status of the patients with self-inflicted gunshot wounds to the face. This study is based on 12 subjects who attempted suicide resulting in extensive maxillofacial injuries using guns placed beneath their chins. The psychiatric evaluation was conducted by interview and using SCID-I, SCID-II, MMPI, Rosenberg Self-Esteem Scale and Suicide Probability Scale. Two subjects were healthy, 1 had bereavement, 6 had current and 5 had previous MDD (major depressive disorder), 2 had dysthymic disorder, 3 had alcohol abuse, 2 had drug abuse and 4 had antisocial personality disorder. The suicidal group was more socially introverted according to MMPI. According to Rosenberg self-esteem subscale, self esteem, the constancy of self respect and depressive mood subtests were statistically significant in the suicide group compared to the healthy controls (P < 0.01). Depressive spectrum disorders are the most common causes. It is obvious that untreated or undiagnosed depression may increase risk of committing suicide. The changes in the physical facial appearance after the suicide attempt caused impairment of self-esteem and the constancy of self respect. Similar to other studies, none of our patients reattempted suicide and all tried to return to their pre-injury lifestyle and appeared to accommodate to the stigma of their physical deformities. Early diagnosis and treatment should be considered as a factor to reduce the risk for suicide attempt. PMID- 17119408 TI - Cranioplasty with auto-purified bone flap after infection. AB - Cranioplasty of cranial bone defects can generally be accomplished with autogeneous bones as well as with nonbiological materials. Autologous living-bone grafts are ideal but require sacrifice of donor bone, and synthetic materials might have possible exposure of the materials, delayed infection, and/or allergic reaction. The authors report cranioplasty with a bone graft after auto purification by subcutaneous preservation of the contaminated bone fragment. A 47 year-old man was given a frontal cranioplasty with a split parietal bone and rib. Epidural abscess ensued, and debridement was performed to control the infection. The largest bone fragment was subcutaneously preserved in the chest wall and reused for cranioplasty. Subcutaneous preservation of bone is a promising strategy for cranioplasty after neurosurgery. The condition of the bone fragment can be inferred from the condition of the site at which it is preserved. If the bone flap is contaminated, it can be purified by the patient's immune system. PMID- 17119409 TI - New therapeutic protocol in the treatment of avascular necrosis of the jaws. AB - Osteonecrosis of the jaws is being increasingly reported in patients with bone metastasis from a variety of solid tumors and disseminated multiple myeloma receiving intravenous bisphosphonates. Agreement exists that these drugs can initiate vascular endothelial cell damage and accelerate disturbances in the microcirculation of the jaws, possibly resulting in thrombosis of nutrient-end arteries. The role of bisphosphonates in cancer patients with previously treated jaws has yet to be elucidated. The signs and symptoms that may occur before the appearance of evident osteonecrosis include changes in the health of periodontal tissues, nonhealing mucosal ulcers, loose teeth and unexplained soft tissue infection. A series of 30 periodontally involved patients showing osteonecrosis of the jaws that appeared following the intravenous use of bisphosphonates is reported. Clinical management of the avascular necrosis of the jaws in patients treated with bisphosphonates presents several problems. An analysis of the international medical literature shows that surgical treatment of the necrotic jaws in patients treated with bisphosphonates has proven to be ineffective in stopping the pathological process. The use of hyperbaric oxygen and antibiotics are not effective, either. The authors have developed a new protocol for the management of these lesions. Compared with other therapeutic choices, this protocol has introduced the use of ozone therapy as therapeutic support. PMID- 17119410 TI - Comparison of infant head shape changes in deformational plagiocephaly following treatment with a cranial remolding orthosis using a noninvasive laser shape digitizer. AB - Deformational Plagiocephaly (DP) is a multi-planar deformity of the cranium occurring either pre-or postnatally in infants. In the last decade, the incidence of DP has increased substantially due to a number of factors, including supine sleeping positioning to reduce Sudden Infant Death Syndrome and the use of child carriers that increase supine positioning. Clinical questions persist about which children should be treated for DP and how to intervene, questions that are difficult to answer without accurate documentation of three-dimensional (3-D) head shape. This study explored a method for quantifying head shape and used that method to evaluate the success of orthotic treatment. Two hundred twenty-four infants who were diagnosed with DP received either a cranial remolding orthosis or a repositioning program with no orthotic intervention. Data from 25 head shape variables were collected using a noninvasive laser shape digitizer. Only variables attributable to growth showed significant differences in the control population, while the treatment population showed significant differences in pre and post-treatment values for all variables. The study identified four variables as particularly important in assessing the head shape of infants with plagiocephaly: the cranial vault asymmetry index, radial symmetry index, posterior symmetry ratio, and overall symmetry ratio. Ninety-six percent or more of subjects in the treatment group showed improvement in each variable. These data document the utility of a 3-D scanning device and the effectiveness of treatment with a cranial remolding orthosis. PMID- 17119411 TI - Calvarial bone grafts and temporalis muscle flap for midfacial reconstruction after maxillary tumor resection: a long-term retrospective evaluation of 17 patients. AB - Midfacial reconstruction after radical oncological resection is a challenging endeavor and several options can be employed: prosthethic devices, pedicled flaps (with or without the aid of autologous or alloplastic grafts), and microvascular flaps. Each technique has specific indications, advantages and disadvantages. The use of traditional surgical reconstructive procedures seems to be shifted nowadays by microvascular free flaps. Nevertheless, in our experience the myofascial temporalis flap associated with free calvarial bone grafts demonstrate to be a safe and versatile option in primary midfacial reconstruction. From this point of view the authors have developed a technique for one-stage reconstruction of the orbito-maxillary skeleton and soft tissues and without the use of microsurgical flaps; this surgical procedure can be used only on patients for whom a resection of the cutaneous tissues and exenteratio orbitae are not necessary. The aims of this paper is to describe the surgical technique and to show a retrospective analysis on 17 patients which underwent midfacial radical resection and immediate reconstruction with calvarial bone grafts and temporalis muscle flap along 15 years. PMID- 17119412 TI - Healing of bone defects by an osteopromotion technique using solvent-dehydrated cortical bone plate: a clinical and radiological study. AB - Osseous defects of the jaws following trauma, cysts, infection or congenital deformity may show poor osteogenesis and the affected area may never be completely filled with bone. It is widely accepted in guided tissue regeneration that physically halting soft connective tissue proliferation into bone allows bone regeneration. This concept is called the "osteopromotion principle." The purpose of the present study was to evaluate the effect of solvent-dehydrated cortical bone plates as a barrier to allow bone regeneration by assessing the osteopromotion principle. 30 patients (18 male, 12 female) with cystic lesions of the jaws were assigned to two different groups. The patients in Group A were treated by enucleation. In Group B, were treated by removal of the lesion and placing a solvent-dehydrated cortical bone plate on top of the cyst cavity to avoid ingrowth of connective tissue. Both groups were evaluated radiographically using spiral-computed tomography (CT) before surgery and 12 months postoperatively. In group B, all cystic cavities showed complete bone healing in 12 months. At the same period, invasion of the soft connective tissue were observed in the patients treated only by enuclation. This study demonstrates that solvent-dehydrated cortical bone plates can be used as a barrier in treatment of cyst cavities and they hinder ingrowth of connective tissues and promote successful bone healing. PMID- 17119413 TI - Branching patterns of the infraorbital nerve and topography within the infraorbital space. AB - The infraorbital nerve (ION) is the terminal branch of the maxillary nerve; it supplies the skin and mucous membranes of the middle portion of the face. This nerve is vulnerable to injury during surgical procedures of the middle face. Severe pain and loss of sense are noted in patients whose infraorbital nerve is damaged. In the study presented here, we investigated the branching pattern and topography of the ION, about which little is currently known, by dissecting 43 hemifaces of Korean cadavers. In most cases, the infraorbital artery was located in the middle (73.8%) and superficial to the ION bundle (73.8%) at its exit from the infraorbital canal. The ION produced four main branches, the inferior palpebral, internal nasal, external nasal, and superior labial branches. The superior labial branch was the largest branch of the ION produced the most sub branches. These sub-branches were divided into the medial and lateral branches depending upon the area that they supplied. We were able to classify four types of branching pattern of the external and internal nasal branch and the medial and lateral sub-branches of the superior labial branch of the ION at the site of their emergence through the infraorbital foramen (types I-IV). Type I, where all four branches are separated occurred the most frequently (42.1%). These findings will help to preserve the ION while performing certain types of maxillofacial surgery, such as removal of a tumor from the upper jaw and fracture of the upper jaw. PMID- 17119414 TI - Innervation of upper orbicularis oris muscle. AB - The aim of this study is to elucidate the innervation of upper orbicularis oris muscle. The upper lips of 17 hemifaces of Korean adult cadavers were dissected. The number of nerve branches crossing and entering the outer margin of pars peripheralis and pars marginalis were counted, respectively. The location of ramifying point of the buccal branch was measured. The nerve branches crossing the outer margin of pars peripheralis were about 4 (3.71 +/- 1.05) on each side of the face and branches crossing the outer margin of pars marginalis was 4.65 +/ 1.46. About four (4.06 +/- 0.83) branches entering the pars marginalis were found. Most ramifying points (14/17, 82%) located within a circle of 5 mm in radius diameter and the center was at 12 mm lateral and 26 mm superior to the mouth corner. An anatomical knowledge might be contributive to understand a wrinkle depletion of the upper lip and function of the orbicularis oris muscle. PMID- 17119415 TI - The relationship of capsulopalpebral fascia with orbital septum of the lower eyelid: an anatomic study under magnification. AB - The aim of this study is to elucidate each insertion of the capsulopalpebral fascia (CPF) and orbital septum (OS) in the lower eyelid. Fifteen eyelid specimens were obtained from 15 Korean male adult cadavers for the study. Three parasagittal sections were made apart at medial limbus, midpupillary line and lateral limbus. The specimens were cut in 10-mum sections, stained with Masson trichrome and observed under light microscopy. The head of the CPF split open superiorly and inferiorly wrapping around the inferior oblique muscle and met anteriorly. CPF inserted to the inferior border of the tarsus, merging the anterior border of the inferior tarsal muscles. OS blended with CPF most closely at 3.7-5.4 mm beneath the lower tarsal border: and differently at 3.7 +/- 0.7 mm on the medial limbus line, 4.3 +/- 0.8 mm on the midpupillary line and 5.4 +/- 1.0 mm on the lateral limbus line. The blending level is closer to the tarsus at the medial side than the lateral side. The closest distance between the lower tarsal border and the first identifiable smooth muscle nuclei of the inferior tarsal muscle was 2.1-2.7 mm, and differently 2.7 +/- 0.7 mm on the medial limbus line, 2.7 +/- 0.7 mm on the midpupillary line and 2.1 +/- 0.9 mm on the lateral limbus line. Precise anatomical understanding of OS and CPF may be conducive to the lower eyelid surgery. PMID- 17119416 TI - Poly-L/D-lactide (PLDLA) 96/4 fibrous implants: histological evaluation in the subcutis of experimental design. AB - Poly-L/D-lactide (PLDLA) 96/4 fibrous implants have been introduced to engineer functional fibrous constructions in situ. The current study was undertaken to evaluate the guidance of the fibrous tissue formation and the tissue reaction of porous PLDLA 96/4 scaffolds implanted in subcutaneous tissue. Three various PLDLA 96/4 knitted-mesh scaffolds (Loose, Ordinary, and Dense) were implanted subcutaneously in 32 rats, and followed-up from 3 days until 48 weeks postsurgery. Histological examination showed that PLDLA 96/4 scaffolds provided a structurally supporting element for 48 weeks. They were filled with fibrous tissue by 3 weeks. During the follow-up, loose connective tissue was organized into dense connective tissue with thick collagen bundles. At 48 weeks, no statistically significant difference was found in the amount of loose or dense connective tissue between the scaffold groups of various porosities, although the tendency for higher amounts of loose connective tissue was seen in the Loose type scaffolds. PLDLA filament diameters were 121 mum at 2 weeks, 119 mum at 24 weeks and 116 mum at 48 weeks (P = 0.03 between 2 and 48 weeks). Porous PLDLA scaffold induced fibrous tissue formation in situ. This can be exploited in engineering fibrous tissue constructs in vivo for tissue support or replacement purposes. PMID- 17119417 TI - Evaluation and management of exposed high-density porous polyethylene implants: an experimental study. AB - Numerous autogenous and alloplastic materials have been used for restoration of contour deformities of the face. Alloplastic materials have come into use to replace autogeneous bone grafts because bone grafts are associated with donor site morbidity, resorption and difficulty in shaping. Porous high-density polyethylene (HDPPE) is a commercial product which is nonallergenic, nonantigenic, noncarcinogenic and nonresorbable, highly stable and somewhat flexible. It has pores of 125-250 mum which enable tissue ingrowth resulting in firm attachment and integration of the implant to the surrounding tissues. In this study we aimed to evaluate the fate of the exposed implants and proposed a method to manage the exposed medpor implants. Twenty rats were used for this experimental study. Three phases involving the implantation, exposition, and the closure of the implants were held respectively. Closure was performed in two groups: 1) After perforating the exposed implants; 2) Without perforating the exposed implants. No serious complications were seen. Perforated exposed implants when covered with flaps rather than grafts could be covered. Exposed implants were not covered easily as indicated by the previous studies. This study demonstrates that early closure of perforated implants with flaps can give the best clinical results when exposed implants are covered in a short period after exposition so that the ingrown fibrous tissues are still intact. We believe that grafting of the exposed medpor implants cannot be effective either in early or late period. Perforating the exposed implants yield successful results as ingrowth of the tissues are enabled. PMID- 17119418 TI - What wrapped perichondrial and periosteal grafts offer as regenerators of new tissue. AB - The major goals in contour restoration procedures are to re-establish the desired contour with the use of resilient and durable materials that can be easily found and harvested. Cartilage grafts are commonly used for these purposes though they often possess a problem of donor site morbidity and shortage of quantity. The neo cartilage formation capacities of both perichondrium and periosteum are well known. We aimed to optimize both the amount and quality of the newly forming tissue from perichondrial and periosteal grafts. For this purpose the grafts were wrapped on themselves. Placement of oxidized regenerated cellulose (ORC) within graft layers was performed in two groups with the aim of giving support to the regenerating tissue, and increasing the connective tissue formation within the graft layers. Three-month-old New Zealand white rabbits were used. Group 1 ear perichondrial, and Group 2 calvarium periosteal grafts of 1.4 x 2.4 cm were harvested, folded on themselves, and sutured at the edges to create closed pockets. 0.8 x 0.8 cm sized ORC sheets were placed inside the pockets before wrapping in Group 3 perichondrial and Group 4 periosteal grafts. 0.2-mL autogenous blood was injected in each pocket. All grafts were transplanted under the abdominal muscle fascia, and harvested after 6 weeks. Volumes and weights of wrapped perichondrial grafts were higher than their periosteal counterparts either with or without the inclusion of ORC. Grafts with ORC (Groups 3 and 4) were heavier than the grafts lacking ORC (Groups 1 and 2), in a statistically significant manner (P /= age 65. At the time of diagnosis, 59% of oral cavity cancers and 29% of lip cancers were at stage III or IV (P < 0.001). The results emphasize an urgent need for a national program focusing on early detection of oral cancers, including educational information addressed to oral health professionals. PMID- 17119437 TI - Nevus sebaceus of Jadassohn. AB - The nevus sebaceus of Jadassohn (SNJ) is a hamartomatous disorder of the skin and its adnexa pertaining to the group of "organoid nevi,'' most frequently involving the face and scalp. During adulthood, patients with SNJ have a 10% to 20% risk of the development of cutaneous or adnexal neoplasia, so that prophylactic excision before puberty is recommended by most authors, and tissue expansion is considered to be the best method of reconstruction. It has been largely demonstrated in literature that most of the lesions that have been interpreted as basal cell carcinoma (BCC) are actually examples of primitive follicular induction or trichoblastomas, not authentic BCCs. A literature review on histopathologic findings associated with SNJ and a retrospective chart review of two cases occurring in young females are presented. In one case, the lesion was treated by intraoperative expander-assisted reduction and scalp graft (Case 1); in the other one, a primary closure with adjacent tissue was performed (Case 2). No signs of malignant degeneration or residual pathology have been found. For treatment of the biggest lesions, when preoperative tissue expansion cannot be performed, intraoperative one, transfer of a scalp graft has been shown to be a good reconstructive method. For the smallest lesions, a primary closure with adjacent tissue is sufficient. PMID- 17119438 TI - Two extreme conditions of xeroderma pigmentosum. AB - Two atypical cases of xeroderma pigmentosum are reported. One of these cases with lung metastasis of squamous cell carcinoma in the first decade of life and the other with late onset disease (XP) in fifth decade of life. Lung metastasis of squamous cell carcinoma in first decade of life in patients with xeroderma pigmentosum has never been reported in the literature to our knowledge. Late onset of the disease in fifth decade is also a rare condition. Clinicians should be more alert for the internal organ metastases to improve the survival of the patients and should never forget that xeroderma pigmentosum has got a wide range of clinical presentation. PMID- 17119439 TI - Genetic effects of Medpor on osteoblast-like cells. AB - Porous polyethylene (PP or Medpor) is an alloplastic material used worldwide for craniofacial reconstruction. Although several clinical studies are available, there is a lack as regard the genetic effects. Because PP is always fixed on bone and the mechanism by which PP acts on osteoblasts is unknown, we therefore attempted to address this question by using microarray techniques to identify genes that are differently regulated in osteoblasts exposed to PP. By using DNA microarrays containing 19,200 genes, we identified in osteoblast-like cell lines (i.e. MG-63) cultured on PP several genes where expression was differentially regulated. The differentially expressed genes cover a broad range of functional activities: 1) signal transduction, 2) transcription, 3) translation, 4) cell cycle regulation, 5) vesicular transport, and 6) production of cytoskeletal elements, cell-adhesion molecules and extracellular matrix components. The data reported are, to our knowledge, the first genetic portrait of osteoblast-like cells cultured on PP. They are relevant to better understanding of the molecular mechanism of bone-PP interaction and as a model for comparing other materials used for bone reconstruction. PMID- 17119440 TI - Novel strategy for orbital tumor resection: surgical "displacement" into the maxillary cavity. AB - Surgical intervention consisting of lateral orbitotomy, the indication of which is extremely wide for orbital tumor surgery, has been applied in cases of large, retrobulbar cavernous hemangioma. However, no method exists involving displacement of the tumor from the crowded orbital contents, with the exception of tumor traction toward the outer side. The impact of traction force on the fragile hemangioma is extremely traumatic and dangerous. The authors examined how a tumor might be "displaced" in the absence of traction force effect, into an appropriate cavity neighboring the orbit. The maxillary sinus may afford the most suitable space to shift the laterally situated orbital tumor. Thus, the osteotomy level was extended to the lateral half of the inferior orbital floor and orbital rim in order to displace the tumor through an "escape window" of sufficient size between the orbit and maxilla. This report describes the treatment of two cases with long histories of progressive proptosis associated with retrobulbar large cavernous hemangiomas. This novel procedure resulted in a successful outcome. The current approach and management, which involves displacement of the tumor into the maxillary sinus through the orbital floor escape window, is a novel procedure for orbital tumor surgery. PMID- 17119441 TI - Aesthetic upper lip reconstruction with vermilion submucosal-pedicle cross-lip flap. AB - The localized cutaneous amyloidosis occurring on the upper lip of a 48-year-old man was surgically treated and aesthetically reconstructed with vermilion submucosal pedicle partial-thickness cross-lip flap. The reconstruction of the upper lip following the tumor resection or trauma with Abbe flap or its modified methods may not be always satisfactory because it leaves parallel scars extending to the free vermilion margin and trapdoor deformity. We considered that the replacement of free margin of the upper vermilion with that of the lower vermilion might be responsible for it. When significant vermilion is lost, some surgeons may discard its free margin additionally or convert this defect to a through-and-through defect for easier repair. We believe that surgeon should preserve a continuity of the free margin of upper-lip if possible. Our modified Abbe flap can be applied for a various defects of the upper lip so far as the free margin of the vermilion can be preserved. PMID- 17119447 TI - NRAS and BRAF mutations in melanoma tumours in relation to clinical characteristics: a study based on mutation screening by pyrosequencing. AB - We have previously demonstrated the use of pyrosequencing to investigate NRAS [neuroblastoma RAS viral (v-ras) oncogene homolog] mutations in melanoma biopsies. Here, we expanded the analysis to include BRAF (V-raf murine sarcoma viral oncogene homolog B1), another member of the Ras-Raf-mitogen-activated protein kinase (MAPK) signalling pathway, and analysed a total of 294 melanoma tumours from 219 patients. Mutations in BRAF exons 11 and 15 were identified in 156 (53%) tumours and NRAS exon 2 mutations in 86 (29%) tumours. Overall, mutations in NRAS or BRAF were found in 242 of 294 tumours (82%) and were found to be mutually exclusive in all but two cases (0.7%). Multiple metastases were analysed in 57 of the cases and mutations were identical in all except three, indicating that BRAF and NRAS mutations occur before metastasis. Association with preexisting nevi was significantly higher in BRAF mutated tumours (P=0.014). In addition, tumours with BRAF mutations showed a significantly more frequent moderate to pronounced infiltration of lymphocytes (P=0.013). NRAS mutations were associated with a significantly higher Clark level of invasion (P=0.022) than BRAF mutations. Age at diagnosis was significantly higher in tumours with NRAS mutations than in those with BRAF mutations (P=0.019). NRAS and BRAF mutations, however, did not influence the overall survival from time of diagnosis (P=0.7). In conclusion, the separate genotypes were associated with differences in several key clinical and pathological parameters, indicating differences in the biology of melanoma tumours with different proto-oncogene mutations. PMID- 17119448 TI - Non-invasive imaging of tissue PO2 in malignant melanoma of the skin. AB - In various tumor systems, decreased PO2 values have been demonstrated by various methods. This study addresses the question of whether tumor hypoxia can be found in cutaneous melanoma using lifetime imaging of non-invasive sensors showing phosphorescence quenched by oxygen. Twenty-three cases of cutaneous malignant melanoma (average tumor thickness 1.25 mm, range 0.5-8 mm) were examined using the SkinCam lifetime imaging system for the assessment of cutaneous PO2 levels within the tumors and in adjacent clinically normal skin. For comparison, 30 non melanoma skin tumors were evaluated. In 15 exploitable melanoma cases, the average hypoxic difference of the lesion compared with the surrounding skin was 10 mmHg, typically associated with an inhomogeneous distribution. Only 10% of the non-melanoma lesions showed a similar hypoxia (false positives). The SkinCam equipment uses a non-invasive imaging method and provides further diagnostic hints in the assessment of benign and malignant skin tumors. PMID- 17119449 TI - Alkylating benzamides with melanoma cytotoxicity: experimental chemotherapy in a mouse melanoma model. AB - The in-vivo antineoplastic potential of the alkylating N-(2 dialkylaminoethyl)benzamides BZA1 and BZA2, novel melanoma targeted anticancer drugs, was evaluated in a mouse melanoma model with nude mice bearing subcutaneous SkMel28, B16 or WM266-4. The maximal tolerated dose (MTD) for the intraperitoneal application of both agents was found to be 24 mg/kg. Treatment was initiated with an intraperitoneal injection of 8 mg/kg of BZA1 or BZA2 on days 0, 2 and 4 in the case of B16 melanoma on days 0, 1 and 2 after the onset of the experiment, when the mean tumor diameter ranged within 4-6 mm. The experiment was terminated when the mean tumor diameter in the control group had reached a value of 12 mm. Tumor growth delay of these agents was compared with dacarbazine (3x250 mg/kg), chlorambucil (3x5 mg/kg) and an untreated control group. Significant tumor growth delay was observed under BZA1, BZA2 and dacarbazine treatment compared with the untreated control group in all three evaluated melanomas with insignificant differences among BZA1, BZA2 and dacarbazine. The insignificant effect of chlorambucil and the strong improvement on growth delay achieved with BZA1 and BZA2 demonstrated melanoma targeting characteristics of the N-(2-dialkylaminoethyl)benzamide structure element. Dacarbazine was more effective in the in-vivo antineoplastic assay compared with the in-vitro cytotoxicity studies, probably due to hepatic bioactivation. Similar side effect intensity of BZA2 and dacarbazine was observed, whereas BZA1 was more toxic. BZA2 might represent an alternative antimelanoma drug, especially in patients not responding to dacarbazine. PMID- 17119450 TI - Malignant melanoma and pregnancy. AB - The occurrence of cancer in pregnant women is not a common phenomenon and the real incidence of malignant melanoma during this period is unknown. Many authors reported a poor prognosis in pregnant women with melanoma compared with non pregnant women's tumour. Several retrospective reviews reported a worsened prognosis in pregnant women with melanoma and found that progesterone and oestrogen receptors can be detected in melanoma tissue. Other data are in conflict with these opinions; several studies demonstrated that the timing of the disease diagnosis during pregnancy did not appear to influence the risk of mortality. In our report, we reviewed data on women with malignant melanoma who were diagnosed during pregnancy in our institute from 1991 to 2000. We have considered the following parameters: age at diagnosis, histological type and tumour thickness, stage of disease and surgical management and we have compared the clinical and biological behaviour of these melanomas with melanoma in non pregnant women observed in the same period and in a follow-up of 5 years. In our study, there is no significant difference in outcome and survival rate between pregnant and non-pregnant women with melanoma. During pregnancy, melanocytic skin lesions show a transient modification of dermoscopic pattern; consequently, a close follow-up of pigmented lesions, both clinical and instrumental, is very important during pregnancy and care must be taken in revealing the presence of other risk factors for melanoma. PMID- 17119451 TI - Decrease of polyamine levels and enhancement of transglutaminase activity in selective reduction of B16-F10 melanoma cell proliferation induced by atrial natriuretic peptide. AB - The atrial natriuretic peptide (ANP) at physiological levels reduced the proliferation of highly metastatic murine (B16-F10) and human (SK-MEL 110) melanoma cell lines whereas rat aortic smooth muscle (RASM) cells were unaffected. In RASM cells, the levels of proliferation markers (putrescine, spermidine and spermine) increase after 24 h of epidermal growth factor (EGF) stimulation (RASM-EGF), but strongly decrease after 24 h of exposition to ANP. The B16-F10 cell line, which received no EGF stimulation, showed a similar decrease in polyamine content after ANP treatment. Furthermore, the enzymatic activity of a differentiation marker (transglutaminase) was increased for both RASM-EGF and B16-F10 cells after 24 h of treatment with 10(-10) mol/l ANP, concomitantly with the observed inhibition of polyamine biosynthesis and cell growth. Data obtained on B16-F10 cells treated with 8Br-GMPc or with an ANP analogue (cANF) support the involvement of the type C ANP receptor (NRP-C) in hormone effects. From the overall results, it appears that ANP may play a role in the inhibition of cellular growth under hyperproliferative conditions, as shown for RASM-EGF cells. The B16-F10 melanoma cell line showed similar results, but in the absence of mitogen stimulation. This observation suggests that the constitutive hyperproliferative state of tumor cells may be a sufficient condition to favor the ANP inhibitory effects on cell growth. This finding is particularly interesting in the light of a possible use of ANP as a potential selective antineoplastic agent. PMID- 17119452 TI - Sodium ascorbate inhibits growth via the induction of cell cycle arrest and apoptosis in human malignant melanoma A375.S2 cells. AB - Vitamin C has been reported to be useful in the treatment and prevention of cancer. Inconsistent effects from growth stimulation to induction of apoptosis of malignant tumor cells, however, have been reported. Melanoma is an increasingly common and potentially lethal malignancy. It was reported that melanoma cells were more susceptible to ascorbate toxicity than any other tumor cells. The mechanisms accounting for ascorbate-induced apoptosis in human melanoma cells, however, have remained unclear. This study was undertaken to investigate the effect of sodium ascorbate on cytotoxicity and apoptosis in human malignant melanoma A375.S2 cells. A375.S2 cells were incubated with a certain range of concentrations of sodium ascorbate for various time periods. In order to examine the effects of sodium ascorbate on cell proliferation, cell cycle, apoptosis and necrosis, we performed 4,6-diamidino-2-phenylindole dihydrochloride assays and flow cytometry analysis. Polymerase chain reaction was used to examine the mRNA levels of p53, p21, p27, cyclin A, cyclin E, CDK2 and CDK4, which are associated with cell cycle S-phase arrest and apoptosis. Flow cytometric analysis showed that sodium ascorbate significantly induced cell cycle arrest and apoptosis in the A375.S2 cell line in a dose-dependent manner. The increased expressions of p53 and p21, and the decreased expressions of cyclin A, cyclin E, CDK2 and CDK4, indicated the cell cycle arrest at G1/S phase after the cells had been treated with sodium ascorbate. Induction of apoptosis involved an increase in the levels of p53, p21 and cellular Ca, and a decrease in mitochondrial membrane potential and activation of caspase 3 before culminating in apoptosis in sodium ascorbate treated A375.S2 cells. PMID- 17119453 TI - Phase 2 trial of Allovectin-7 in advanced metastatic melanoma. AB - Treatment of metastatic melanoma with chemotherapeutic regimens has led to disappointing response rates, duration of response and no appreciable impact on survival. Intralesional injection of a low dose of an HLA-B7/beta2 microglobulin plasmid formulated with cationic lipids (Allovectin-7 is a registered trademark of Vical, Incorporated, San Diego, California, USA) has been shown previously to be safe and well tolerated. A phase 2, open-label study was performed at 16 centers in the United States. Seventy-seven patients were treated with 10 mug intralesional Allovectin-7 weekly for 6 weeks and clinical response to treatment were evaluated by World Health Organization criteria. Minimal adverse events were associated with the Allovectin-7 injections. Seven patients (9.1%) had complete or partial response with 4.8 months median duration of response. Allovectin-7 was shown to be safe and exhibit biological activity at this dose. Its safety profile may enable Allovectin-7 to be used at higher doses, which may provide greater clinical activity. PMID- 17119454 TI - The altered apoptotic pathways in cisplatin and etoposide-resistant melanoma cells are drug specific. AB - Apoptotic deficiency is one of the mechanisms leading to chemoresistance due to the potential of many chemotherapeutic drugs to induce apoptosis. We have examined drug-induced apoptosis in the chemosensitive human melanoma cell line MeWo, as well as in its resistant sublines, which were selected by continuous exposure to etoposide (MeWo(Eto1)) and cisplatin (MeWo(Cis1)). In former studies, activation of the mitochondrial pro-apoptotic pathway could not be demonstrated in etoposide-resistant cells after exposure to etoposide. A significant reduction of PARP [poly (ADP-ribose) polymerase] cleavage and caspase activation, but unimpaired DNA fragmentation, was seen in cisplatin-resistant cells after treatment with cisplatin. In the current study, we investigated effects of chemotherapeutic drugs different from the selecting agents cisplatin and etoposide on the observed modulations of the mitochondrial apoptotic pathway. We analysed dose-dependent release of cytochrome c, caspase-9 activation, cleavage of PARP and activation of effector caspases in etoposide and cisplatin-resistant cells after exposure to etoposide, teniposide, cisplatin or fotemustine. In analogy to etoposide exposure, we could not demonstrate any activation of the apoptotic pathway in etoposide-resistant cells after exposure to teniposide, another topoisomerase-II inhibitor. In contrast, exposure to cisplatin and fotemustine led to apoptotic cell death in these cells. This suggests that the deficiency of apoptosis in etoposide-resistant cells is dependent on the trigger by topoisomerase-II inhibitors. Analysis of cisplatin-resistant cells after etoposide and fotemustine exposure revealed an increased activity of the apoptotic pathway when compared with cisplatin exposure at corresponding survival rates in these cells. These results suggest that the observed modulations of the apoptotic pathway in resistant melanoma cell lines are specific for an anti neoplastic drug and are not fixed at the molecular level, as different chemotherapeutic drugs are capable of overcoming these alterations. PMID- 17119455 TI - The effect of blue light exposure and use of intraocular lenses on human uveal melanoma cell lines. AB - Little is known about the effect of blue light on inducing melanocytic malignant transformation. We chose to investigate the effect of blue light (475 nm wavelength) on the proliferation rates of uveal melanoma cells. In addition, we tested two different intraocular lenses to determine the possible effects of ultraviolet absorbing and blue light filtering intraocular lenses on the changes in proliferation. Four human uveal melanoma cell lines (92.1, MKT-BR, OCM-1, SP6.5) were exposed to blue light with and without the presence of ultraviolet absorbing and blue light filtering intraocular lenses. Cells covered by aluminum foil were used as a control. The proliferation rate of the cells compared with the control was then assessed using the Sulforhodamine-B proliferation assay. Cells exposed to blue light showed a statistically significant (P<0.05) increase in proliferation. Those exposed to blue light through a standard ultraviolet absorbing intraocular lens showed a smaller increase in proliferation, whereas those exposed with a blue light filtering intraocular lens showed no increase in proliferation than the control in all four cell lines. The exposure of cells to blue light led to an increase in proliferation in all cell lines compared with the control. The use of blue light filtering intraocular lenses abolished these increases in proliferation in the four cell lines. These results indicate that blue light filtering intraocular lenses may have a protective effect on the proliferation rates of uveal melanoma cells exposed to blue light. PMID- 17119456 TI - Persistent thrombocytopenia during melanoma treatment with fotemustine. AB - Haematological toxicity characterized by delayed and reversible neutropenia and/or thrombocytopenia is an adverse effect observed in 40% of patients receiving fotemustine. We report the case of a 66-year-old man with metastatic malignant melanoma treated with fotemustine as monotherapy. A severe persistent and prolonged thrombocytopenia occurred, so that chemotherapy was discontinued. Bone marrow involvement was excluded. The physician should be aware of this prolonged thrombocytopenia secondary to fotemustine, so that it may be recognized early and not attributed erroneously to tumour evolution. This observation could be substantial for the design of combination regimens with fotemustine and other myelotoxic drugs in pretreated melanoma patients. PMID- 17119457 TI - Acetabular outcome after hip hemiarthroplasty in patients with tumors. AB - Hip hemiarthroplasty is performed routinely on patients with tumors of the proximal femur, although the long-term effect on the native acetabulum is unknown. We measured the amount of femoral head migration that would occur with intermediate and long-term followup. We also measured the amount of groin pain experienced by patients with longer followup and the overall rate of conversion to total hip arthroplasty. From a surgical database, we identified 442 patients with tumors who had 447 hip hemiarthroplasties without or with an allograft composite. We reviewed the medical records and radiographs of 32 patients with at least 5 years followup (median followup, 10 years) to determine their clinical and radiographic outcomes. The median proximal and medial migration measurements were 3 mm (range, 0-24 mm) and 2 mm (range 0-20 mm), respectively. Thirty-one patients had minimal or no groin pain. Seven of the 447 arthroplasties (1.6%) were converted to total hip arthroplasties. Even patients with long-term followup usually had minimal pain and radiographic changes. The overall rate of conversion to total hip arthroplasty was low. Concern regarding excessive acetabular wear resulting in conversion to total hip arthroplasty in patients with tumors is not supported by our data. PMID- 17119458 TI - Platelet and leukocyte activation in salvaged blood and the effect of its reinfusion on the circulating blood. AB - Postoperative wound drainage reinfusion reduces the frequency of homologous blood transfusion. The salvaged blood is depleted of coagulation factors but may contain platelets and leukocytes which are activated, and therefore potentially procoagulant. We ascertained the degree of activation of platelets and leukocytes in salvaged blood and asked whether their infusion produced any measurable effect on patients' coagulation system. We prospectively randomized 24 patients who had total knee arthroplasties to reinfusion of salvaged autologous blood (n = 12) or a standard drain with no reinfusion (n = 12). Analysis of the salvaged blood showed marked activation of platelets as shown by their expression of P-selectin, CD40 ligand, and Factor V/Va, and as increased numbers of platelet-derived microparticles. After reinfusion there was no measurable effect on activation markers of circulating platelets or leukocytes but there was a decrease in platelet count in the reinfused group compared with the control group. Levels of prothrombin fragment F1 + 2 (suggesting thrombin formation) increased in the reinfused group compared with control group, possibly indicating activation of coagulation systemically. The platelets and leukocytes in salvaged blood are markedly activated and their reinfusion causes a decrease in platelet count in the recipient and a possible increase in thrombin generation potentially favoring thrombosis. PMID- 17119459 TI - Femoral remodeling may influence patient outcomes in slipped capital femoral epiphysis. AB - Clinical studies of patients treated for slipped capital femoral epiphysis have found limited functional impairment and femoral neck deformity causing eventual coxarthrosis. Since patient-focused assessments minimize bias and reflect health related quality of life status, we coupled their use to a clinical examination to obtain a more patient-centered picture of slipped capital femoral epiphyseal outcomes. The impact of residual deformity on outcomes also was examined. Of 78 patients treated for slipped capital femoral epiphyses between 1972 and 1998, 29 (38 hips) were evaluated at a mean followup of 7.6 years (range, 1.4-26 years). The average patient age was 21.8 years (range, 14.6-39 years), 55% were female, and the average body mass index was 28.7 (range, 16.1-50.2). Most slips were stable (92%, 35 of 38) and mild or moderate in severity (98%, 36 of 37). Followup examinations revealed slight deficits in range of motion, strength, and limb length. Radiographs showed slight improvements in head-shaft angle and reduced but persistent femoral neck deformity. Osteoarthritic changes were absent or negligible in 84% (32 of 38) of the hips. The average Iowa hip score was 90.5 (range, 51-100). Patient outcome scores for the AAOS Hip/Knee Questionnaire fell slightly below 50th percentile norms. Neither slip stability, severity, nor body mass index impacted outcome. Femoral neck deformity correlated with function, pain, and Boyer grade. Overall, patients had minor functional deficits and pain that may have been related to femoral neck deformity, but longer followup is warranted. PMID- 17119460 TI - Popliteal mass with knee pain in a 57-year-old woman. PMID- 17119461 TI - Arthroscopic management of labral tears in the hip: a systematic review of the literature. AB - Arthroscopic management of labral problems in the hip has become an accepted therapeutic modality in appropriately selected patients. We performed a systematic review of the literature to determine the rate of patient satisfaction that can be expected following acetabular labral debridement. Computerized literature databases were searched from January 1980 to September 2005 to identify relevant articles that met inclusion criteria and had at least 2 years followup. We included patients with symptomatic acetabular labral tears who failed conservative management, were not claiming workers' compensation, and did not have severe arthritis or severe acetabular dysplasia. Following labral debridement this patient population can expect: (1) a patient satisfaction rate of approximately 67% at 3.5 years follow-up; (2) good results by a modified Harris Hip Score in patients who are subjectively satisfied with their outcome; and (3) a complete resolution of mechanical symptoms in nearly 50% of patients with this complaint. Although limited, the current literature supports non workers' compensation patients with isolated labral tears, who lack associated intraarticular abnormality, can receive both symptomatic and functional improvement following arthroscopic labral debridement. PMID- 17119462 TI - A lytic bone lesion in the phalanx of an 18-year-old man. PMID- 17119464 TI - [Introduction: from molecular oxygen to oxidative stress and radical biochemistry]. PMID- 17119463 TI - Using navigation to measure rotation kinematics during ACL reconstruction. AB - Rotational kinematics of the knee is not fully restored after single-bundle anterior cruciate ligament (ACL) reconstruction. Cadaveric experiments using knee testing machines have suggested anatomical reconstruction replacing the anteromedial and posterolateral bundles could restore knee kinematics more effectively than single-bundle reconstruction. However, practical tools to objectively assess knee rotational laxities clinically have not been available. We used an optically based computer-assisted navigation system to measure the tibiofemoral motion kinematics in four fresh whole cadavers. Standard clinical knee laxity tests (anterior drawer, Lachman, and pivot shift) were performed and the kinematics described in terms of tibial axial rotation and anteroposterior translation. Data were obtained for intact knees after excision of the ACL and sequential reconstruction of the anteromedial and posterolateral bundles. In the ACL-deficient knee, the mean maximum tibial rotation during the pivot shift test was 27 degrees and mean maximum translation 11 mm. Reconstruction of the anteromedial bundle reduced the rotational component to 18 degrees and translation to 7 mm. Reconstruction of the posterolateral bundle reduced rotation to 14 degrees . This pilot study suggests computer assisted navigation could provide a practical method to objectively measure the pivot shift and may be used clinically to demonstrate differences in the control of tibiofemoral rotation kinematics afforded by single and two-bundle ACL reconstructions. PMID- 17119465 TI - [Physico-chemical aspects of reactive oxygen species]. AB - Reactive Oxygen Species (ROS), namely hydroxyl (*OH) and superoxide (O2*-) free radicals and hydrogen peroxide (H2O2), are involved in all oxidative stress phenomena. These latter occur in numerous pathological disorders such as, for example, cardiovascular diseases, diabetes or neurodegenerative diseases. Knowledge of thermodynamic (reduction potentials) and kinetic (rate constants) properties of ROS allows to draw up a rigorous overview of the chemical reactivity of these species. Hydroxyl free radicals (*OH) are powerful oxidants (very toxic species) which attack all the biomolecular targets (DNA, proteins, lipids...), giving other free radicals localized on the targets. Superoxide free radicals (O2*-) have a more graduated reactivity, since they don't directly react with biological molecules. However, some toxicity would be attributed to them via secondary radicular reactions. Other free radicals (of peroxyl RO2* and alkoxyl RO* types), belonging also to the ROS family, contribute by enhancement to oxidative stress. PMID- 17119466 TI - [Cellular sources of reactive oxygen and nitrogen species. Roles in signal transcription pathways]. AB - The history of studies regarding reactive oxygen and nitrogen species (ROS/RNS) is approximatively of 50 years. ROS were shown initially for their deleterious effects on marcormolecules such as DNA and proteins, leading to deterioration of cellular functions as an oxidative stress. On the other hand, recent studies have demonstrated that ROS/RNS act as oxidative signalling in cells, resulting in various gene expressions. This brief review focuses on the main cellular origins of ERO/ERN, such as mitochondrial respiratory chain, NAD(P)H oxidase and NO synthases, and describe the modulation by the reactive species of two major signal transduction pathways, NF-KB and AP-1 pathways. PMID- 17119467 TI - [Oxidative stress and damages to biomolecules (lipids, proteins, DNA)]. AB - Reactive oxygen species or peroxynitrite, coming from the reaction of nitric oxide and superoxide anion, are strong oxidants capable of damaging lipids, proteins and DNA. The oxidative products issuing from each biomolecule are complex and multiple. Reactivity, the mechanism of production and the products formed vary depending on the free radical (superoxide anion, hydroxyl radical, peroxynitrite) and the molecular target (phospholipids, cholesterol, aromatic or aliphatic amino acids, puric or pyrimidic bases). Some of these oxidative products are markers of oxidative stress. For example malondialdehyde and isoprostanes are oxidative markers of lipids, carbonylated proteins of proteins and 8-oxo-guanine or 8-oxodeoxyguanosine of DNA. However other products are also produced, as is the case of the reaction of peroxynitrite with tyrosine which leads to the formation of 3-nitrotyrosine. The quantification of 3-nitrotyrosine is labor-intensive and requires specific equipment. The major problem when searching for the most appropriate marker for a given disease is the great diversity of oxidative products formed depending on the nature of the free radical involved. PMID- 17119468 TI - [Oxidative stress in human diseases]. AB - Oxidative stress is an abnormal phenomenon occurring inside our cells or tissues when production of oxygen radicals exceeds their antioxidant capacity. Excess of free radicals damage essential macromolecules of the cell, leading to abnormal gene expression, disturbance in receptor activity, proliferation or cell dye, immunity perturbation, mutagenesis, protein or lipofushin deposition. Numerous human diseases involve during the pathological process such a stress, localized or general (in the same way as inflammation). In many serious diseases such as cancer, ocular degeneration (age related macular degeneration or cataract), neurodegenerative diseases (ataxia, amyotrophic lateral sclerosis, Alzheimer's disease) stress is the factor original. In familial amyotrophic lateral sclerosis the genetic abnormality occurred an abnormal coding for an antioxidant enzyme, copper-zinc super oxide dismutase. In various other diseases oxidative stress occur secondary to the initial disease but plays an important in role immune or vascular complications. This is the case in infectious disease such as AIDS or septic shock, Parkinson's disease or renal failure. So antioxidant treatment seems logical to be tested in these pathologies. But they have to be applied early in the process, before irreversible mechanisms. They need also to be prescribed at low doses as baseline free radical production have to be preserved to maintain useful activity that cannot be suppressed. PMID- 17119469 TI - [The SU.VI.MAX study, a randomized, placebo-controlled trial on the effects of antioxidant vitamins and minerals on health]. AB - The SU.VI.MAX study is a double blind, randomized, placebo-controlled trial testing, for 7,5 years, the effect of a combination of antioxidant vitamins and minerals, at doses considered to be nutritional (120 mg vitamin C, 30 mg vitamin E, 6 mg beta-carotene, 100 microg selenium and 20 mg zinc) in reducing cancer and ischemic vascular disease incidence in a general population (12.741 middle-aged). After 7.5 years, low-dose antioxidant supplementation had no effect on vascular disease incidence. This dose lowered, however, total cancer incidence in men, but not in women. With regard to contradictory results of observational and interventional studies published for the last decades, we can consider that the effect of antioxidants on cancer may depend on the doses (nutritional versus pharmacological), baseline antioxidant status (different between gender and/or nutritional status) and health status of subjects (healthy versus cancer high risk subjects). Antioxidant supplementation may have a beneficial effect on cancer incidence only in healthy subjects who are not exposed to cancer risk, and with a particularly low baseline status. Finally, antioxidants as well as free radicals appear to be ambiguous nutrients with a wide range of benefits and toxicity. High doses of antioxidant supplements may be deleterious in high-risk subjects without any clinical symptoms in whom the initial phase of cancer development has already started. PMID- 17119470 TI - [Conclusions, future prospects and recommendations]. AB - Understanding the notion of oxidative stress implies a good knowledge of the reactivity of the different reactive oxygen species (ROS). Cell damage can be induced by an overproduction of these species and/or by a deficiency in the protective antioxidant systems. Nevertheless, ROS do not display only deleterious effects and play key-roles in cell signalisation and regulation of the expression of redox sensitive genes. Besides ROS, reactive nitrogen species (RNS) with nitric oxide (*NO) as leader element, are widely involved in biology and lead to the term "nitrosative stress" that particularly describes the damage induced by peroxynitrite, a species formed by reaction between superoxide anion and degrees NO. Nutritional strategies have been based on antioxidant-rich diets, or on supplementation with antioxidants; they could constitute adjunct therapies of interest. Given all these data, radical biochemistry must be considered as a specific discipline. PMID- 17119471 TI - [Therapeutic failure: importance of genes?]. AB - Drug management can be a difficult task in certain situations because of the variable response observed from one patient to another. Genetic factors affecting the pharmacokinetics and pharmacodynamics of drug reactions could explain the interindividual variability in drug response. Pharmacogenetic analysis provides insight into the molecular mechanisms involved in drug response, with the ultimate goal of achieving optimal drug efficacy and safety. Numerous polymorphisms have been described in genes encoding drug-metabolising enzymes, transporters, and receptors. For some drugs, the impact on drug bioavailability and effect has been elucidated. We review here the molecular basis of interindividual variation in drug response and the methods used to identify individual risk of drug failure or toxicity. Clinical applications, concerning enzymes metabolising drugs (cytochrome P4502D6, thiopurine S-methyltransferase and N-acetyltransferase) provide an illustrative demonstration of the usefulness of pharmacogenetic tests in improving patient management. Clinical validation of these tests and new technologies (real-time PCR, DNA chips) should, in the future promote pharmacogenetics in clinical practice and may be lead to more individualized drug therapy. PMID- 17119472 TI - [John Vane, 1927-2004, the pharmacologist of the vascular endothelium [corrected]]. AB - The work of John Vane greatly contributed to the use of aspirin in cardiology. The impact of aspirin administration at low dose for the prevention of stroke or coronary attack results from its effect on enzymes regulating the production of prostaglandins. After understanding the mechanism of interaction between aspirin and the vascular endothelium, Vane proposed assigning a major physiological function to the vascular endothelium which then became a pharmacological target for new drugs. Using an ingenious "real time" biological assay of bloodstream hormones irrigating an isolated organ called the "blood-bathed organ cascade", John Vane developed a system for highly sensitive monitoring of several mediators like angiotensin, bradykinin and prostaglandins and discovered prostacyclin, a potent platelet aggregation inhibitor. Vane explained anti-inflammatory drugs effects (among which aspirin remains the leader) through their activity on cyclo oxygenase and inhibition of prostacyclin and thromboxane production. Another cyclooxygenase isoform, so-called type 2, has been discovered in 1991. Thus, besides the constitutive COX-1, participating to stomach protection and renal artery vasodilatation, a COX-2 enzyme is existing, induced by inflammatory phenomenons and cytokines stimulation, allowing to design specific inhibitors "coxibs", playing an increasing but controversial role in the struggle against inflammation. He won Albert Lasker Prize in 1977 and Nobel Prize in medicine and physiology (shared with Sune Bergstrom and Bengt I. Samuelson) in 1982. PMID- 17119473 TI - [Chronobiology]. PMID- 17119474 TI - [Disruption of biological rhythms and mood disorders]. PMID- 17119475 TI - [The melatonin system, biological rhythms and mood disorders]. PMID- 17119476 TI - [Biological rhythms and anxiety disorders]. PMID- 17119477 TI - [Disruptions of circadian rhythm in neurologic disorders]. PMID- 17119478 TI - [Interactions of melatonin with the central nervous system]. PMID- 17119479 TI - [The melatoninergic system]. PMID- 17119480 TI - [Synchronization and desynchronization of the biological clock in man]. PMID- 17119481 TI - [Establishment of biological rhythms in the course of development]. PMID- 17119482 TI - [Change in rhythms with age]. PMID- 17119483 TI - [Consensus of experts of the WPA (World Psychiatric Association) on atypical antipsychotics]. PMID- 17119484 TI - [Evolution of international consensus on antipsychotic agents of the second generation]. PMID- 17119485 TI - [Methodology of consensus]. PMID- 17119486 TI - [Therapeutic guidelines in schizophrenias]. PMID- 17119487 TI - [The course of the drug, clinical research and commercialization. Constraints, obstacles, delays and costs]. PMID- 17119488 TI - [Evidence data and importance of public health of the new antipsychotics]. PMID- 17119489 TI - [Report of the working group "families and patients"]. PMID- 17119490 TI - [Report of the working group: the place of nonmedical caregivers]. PMID- 17119491 TI - [The routine prescription: between references and reality]. PMID- 17119492 TI - [4th meeting in psychiatry: schizophrenias today]. PMID- 17119493 TI - [Neurodevelopmental hypothesis of schizophrenia]. PMID- 17119495 TI - [Gene/environment interactions in schizophrenia]. PMID- 17119497 TI - [Relevance of cognitive tools for early diagnosis of schizophrenia]. PMID- 17119499 TI - [Schizophrenia and manic depressive disorder: current studies of a unified hypothesis]. PMID- 17119501 TI - [Do cognitive sciences open therapeutic paths in schizophrenia?]. PMID- 17119503 TI - [The dopaminergic hypothesis revisited]. PMID- 17119505 TI - [Non-dopaminergic drugs in schizophrenia]. PMID- 17119507 TI - [Preventive treatments in schizophrenia]. PMID- 17119509 TI - [Resistant schizophrenia: definitions and procedures]. PMID- 17119510 TI - [Efficacy and tolerance of antipsychotic agents: what are the current issues?]. PMID- 17119511 TI - [How to improve compliance in the patients with schizophrenia?]. PMID- 17119514 TI - Physiological characteristics of elite professional soccer teams of different ranking. AB - AIM: Few studies have examined the role of physiological characteristics of soccer teams in the finishing position in the league. The purpose of the present study was to investigate whether there are differences in anthropometric characteristics, endurance, isokinetic peak torque, vertical jump height among professional soccer teams with different rank in the Greek championship. METHODS: Three soccer teams that participated in the Greek Championship participated voluntarily in the present study. The B (n=19; 26+/-4 years; 180+/-5 cm; 78+/-4.5 kg) team was among the best three teams of the National Greek league, whereas M (n=15; 24+/-4 years; 178+/-4 cm; 74.8+/-4.2 kg) and L (n=20; 23+/-3 years; 179+/ 7 cm; 75.3+/-6.4 kg) teams were in the middle and last of the rank, respectively. All teams were evaluated in body fat (%), peak torque of knee extensors at 1.05 rad/s, running velocity at the lactate threshold (LT) and countermovement jump height. RESULTS: The B team, which was deemed among the best three team of the league, presented significantly (P<0.05) lower body fat (%) values, and higher peak torque of knee extensors, running velocity at the LT, and vertical jump height values in comparison to the middle and last team of the league. CONCLUSIONS: The findings of the present study suggest that the physiological characteristics may play an important role for high soccer performance, as it is reflected from the rank order placed in the Greek championship. Especially, body fat (%), running velocity at the LT, peak torque of knee extensors and vertical jump ability could be differentiating factors for a soccer performance. PMID- 17119515 TI - Seasonal variation of aerobic performance in soccer players according to positional role. AB - AIM: The purpose of this study was to evaluate the seasonal variation in the cardiorespiratory fitness of soccer players according to positional role in the team. METHODS: Thirty-two male soccer players U-19 years old were studied, consisting of 10 central defenders (18.2+/-0.9 years and 8.5+/-1.6 years of training (Group A), 12 midfielders (18+/-1 years and 8.1+/-1.4 years of training (Group B) and 10 attackers (18.2+/-0.9 years and 8.4+/-1.5 years of training (Group C). All subjects underwent anthropometric measurements and maximal exercise tests on a treadmill four times during their annual training cycle and maximal blood lactate accumulation analysis in order to verify that maximal exertion was reached. The measurements took place in the beginning and at the end of the preparation period, in the middle as well as at the end of the competitive season. RESULTS: Group A showed increased VO2(max) (mL.kg(-1.)min(-1)) by 19.3% in the 2nd measurement, by 22.9% in the 3rd and by 17.6% in the 4th compared to the beginning, respectively (P ET>shorts (P<0.001). CONCLUSIONS: CT compared to ET have positive effects on calf muscle oxygenation and venous pooling in quiet resting positions. PMID- 17119520 TI - Creatine supplementation does not impair the thermoregulatory response during a bout of exercise in the heat. AB - AIM: The aim of this work was to examine the effects of creatine (Cr) supplementation on resting body water volumes and on core temperature and sweat loss during a bout of exercise in a warm environment. METHODS: Twenty-four aerobically trained male subjects (age 22.93+/-3.01 years, height 179.52+/-7.28 cm, mass 82.06+/-14.32 kg) volunteered to participate in this study. Each subject was assessed for resting body water volumes and for body mass (BM), heart rate (HR), blood pressure (BP), and core temperature immediately before and following a 60-min bout of exercise in a warm environment. Core temperature, HR, and BP were also recorded at 10-min intervals during exercise. Subjects were then randomly assigned to either a Cr or placebo (P) group. Each subject returned following a 5-day supplementation period and was reassessed using identical testing procedures. BM was measured using a standard electronic scale. Body water volumes were assessed using a multi-frequency BIS (Xitron Technologies, San Diego, CA, USA). Core body temperature was measured using a CorTemp Disposable Temperature Sensor and a CT2000 Miniaturized Ambulatory Recorder (HTI Technologies, Inc., Palmetto, FL, USA). RESULTS: The Cr group experienced a significant increase in all body water volumes. No changes were observed in the P group. No changes in core temperature or sweat loss were observed in either group following supplementation. CONCLUSIONS: Cr loading did not impair the thermoregulatory response during a bout of exercise in the heat. PMID- 17119521 TI - Aerobic exercise in water versus walking on land: effects on indices of fat reduction and weight loss of obese women. AB - AIM: It has been suggested, that water exercise is less effective than weight bearing exercise on land for body fat reduction. METHODS: To test this hypothesis 38 middle-aged obese women (25-47% body fat) participated in a 13 week exercise diet program to compare the effects of aerobic exercise in water versus walking on land on indices of fat reduction and weight loss changes. Subjects were randomly assigned to 1 of 3 exercise groups: 1) walking on land (WL), 2) swimming (SW) at 27 degrees C water temperature and 3) walking in 29 degrees C water (WW) at the shallow end of a declining pool with the water at navel height. Subjects in the SW group alternated breast-, side-, and backstroke swimming without face immersion. Exercise parameters were kept constant for all three groups. Subjects participated in supervised exercise sessions for 40 min, 4 times a week at 70% of age-predicted maximum heart rate. Subjects were tested before and after the 13 week experimental period. RESULTS: Significant reductions in body weight, (5.9 kg), percent body fat, (3.7%), and skinfold and girth measurements, occurred in all groups. There where no significant differences between groups. CONCLUSIONS: The results of this study indicate that there are no differences in the effect of aerobic activities in the water versus weight-bearing aerobic exercise on land on body composition components as long as similar intensity, duration and frequency are used. PMID- 17119522 TI - Cumulative effects of conjugated linoleic acid and exercise on endurance development, body composition, serum leptin and insulin levels. AB - AIM: The aim of this study was to evaluate the accruing effects of conjugated linoleic acid (CLA) supplementation and aerobic exercise (Ex) on body composition and serum lipid profile in humans. METHODS: Forty-four healthy female young subjects were divided ExCLA, CLA, Ex and control (C) groups. The groups of CLA and ExCLA were supplemented with 3.6 g/d CLA whilst ExCLA and Ex groups were exercised for 30 min(-1.)3 days(-1.)week(-1) for 6 weeks. RESULTS: After the study period, fat ratio, fat mass, waist and hip girths were reduced in all experimental groups and fat-free mass induced in ExCLA and CLA groups and body weight was reduced in the CLA group when compared to baseline levels. These alterations were significantly different than those of controls with the highest variations were observed in the ExCLA. There was no change in serum leptin, apo AI, apo-B, total cholesterol, HDL, LDL, free fatty acid, and triglyceride levels. Serum glucose concentrations of ExCLA and CLA groups and insulin level of ExCLA group decreased significantly as compared to baseline levels with only serum glucose reduction of both groups were significantly different than those of controls (P<0.05). Endurance performance significantly increased in ExCLA and Ex groups (P<0.01) but did not vary in the CLA and controls. CONCLUSIONS: It was shown that both CLA and exercise were effective in improvement of body composition and these effects were cumulated when they have been used together. CLA supplementation alone or with exercise seems effective on serum glucose and insulin concentrations but ineffective on endurance performance. PMID- 17119523 TI - Positive short-term subjective effect of sports drink supplementation during recovery. AB - AIM: The aim of this study was to investigate the effects of a naturally composed sports drink containing proteins and carbohydrates used during recovery in competitive badminton players. The hypothesis was that the use of a recovery drink would lead to positive subjective effects, enhanced physical performance and less signs of overtraining. METHODS: During an in-door season 18 badminton players were instructed to drink at least 250 mL of a given sports drink immediately after each training or playing session. The study design was prospective double blind crossover with one active drink and one placebo. The active drink was based on natural products containing whey and orange juice, and the placebo was made of diluted apple juice. Evaluation of effects was done with laboratory tests, self-registered values and field tests. RESULTS: The players perceived statistically significant short-term subjective positive effects after using the active drink, compared with after using placebo. The blood hemoglobin concentration was also higher after the period with active drink. There were no other differences concerning other laboratory tests (leg strength, endurance, body fat percent, lean arm and leg masses), self-registered values (body weight, pulse, training amount and intensity) or field tests (speed, explosive effort, grip strength, endurance and POMS) between the periods with the different sports drinks. CONCLUSIONS: Supplementation with a sports drink during recovery showed a significant short-term subjective positive effect compared with placebo. However, no effects were seen on physical performance or signs of overtraining. PMID- 17119524 TI - The reliability of isokinetic and isometric leg strength measures among individuals with symptoms of mild osteoarthritis. AB - AIM: The aim of the study is to evaluate the test-retest reliability of measures of isokinetic and isometric leg strength and joint function among individuals exhibiting symptoms of mild osteoarthritis. Reliable procedures are needed to assess the effectiveness of an intervention on osteoarthritic symptoms. METHODS: Test-retest reliability of two leg strength protocols was assessed using the intraclass correlation coefficient (R). Testing was completed on two occasions separated by 7 days. Eighteen subjects (9 male and 9 female; 54.1+/-11 years) completed an isokinetic testing trial, which consisted of a set of 5 maximal repetitions of the quadriceps and hamstrings at 60 deg/s followed by a set of 15 maximal contractions at 180 deg/s with a 2-min rest between sets and an isometric testing trial, which consist of 3 maximal contractions of the quadriceps for 6 s with a 30-s rest between contractions at 30, 45, and 80 degrees of knee flexion for a total of 9 isometric contractions. A 90-s rest occurred between angles. RESULTS: Most of the isokinetic variables showed moderate to high intraclass reliability (ICC). Two of the calculated isokinetic variables (work fatigue at 180 degrees /s for extension and for flexion) showed low intraclass reliability (ICC=0.78, resp. ICC=0.6). All calculated ICC values of the isometric variables were moderate to high. CONCLUSIONS: Test-retest reliability of isokinetic and isometric leg strength was high, allowing the intervention protocol to monitor changes in leg strength and joint function among those exhibiting symptoms of mild osteoarthritis. PMID- 17119525 TI - Differences in exercising limb blood flow variability between cardiac and muscle contraction cycle related analysis during dynamic knee extensor. AB - AIM: Blood flow in peripheral conduit arteries during steady-state, dynamic exercise, can be estimated noninvasively with Doppler ultrasound, by measuring the conduit arterial diameter and the mean blood velocity averaged over consecutive cardiac beat-by-beat cycles (BB(cycle)) or muscle contraction relaxation cycles (CR(cycle)). The precise impact fluctuations in the 1-BB(cycle) or 1-CR(cycle)-rate may impose on the average blood flow measurements has previously not been clearly defined. The hypothesis investigated in the present study was that the blood flow measurements obtained, and its variability, during exercise, may differ between the 1-BB(cycle) and 1-CR(cycle) at incremental exercise intensities; as the BB(cycle)-measurements may be influenced by transient alterations in heart rate; whereas the CR(cycle)-measurements are dependent on the muscle contraction-relaxation frequencies independent of the exercise intensities per se. The main purpose was therefore to determine if fluctuations in blood flow for 1-BB(cycle) and 1-CR(cycle)varies at incremental exercise intensities (work rates) using the one-legged dynamic knee-extensor exercise (DKE) model. METHODS: Limb femoral artery blood flow (LBF) was determined, for 1-BB(cycle) and 1-CR(cycle), in 8 healthy male subjects during 4 min of steady-state DKE at 60 contractions per minute at 10, 20, 30 and 40 W. The variability of LBF was determined from the coefficients of variation (CVLBF). RESULTS: The CV(LBF) for the CR(cycle)-measurements at each work rate were similar (P=NS). The CV(LBF) for the BB(cycle)-measurements were higher (P<0.05) at 40 W compared to at 10 W. Furthermore, the CV(LBF) for the 1-BB(cycle) was higher (P<0.05) than for the 1-CR(cycle) at 30 and 40 W, despite almost identical mean LBF values for the BB(cycle)- and the CR(cycle)-measurements at each exercise intensity. CONCLUSIONS: The present data suggests that estimates of LBF at slightly higher exercise intensities such as above 30 W, for a few number of consecutive BB(cycle), renders a higher variability than for CR(cycle) measurements. This may consequently result in slight over- and under-estimations of LBF compared to the CR(cycle)-measurement. PMID- 17119526 TI - Autonomic and psychological adaptations in Olympic rowers. AB - AIM: Power spectral analysis of heart rate variability has been used to assess the time course of neurovegetative adaptations to training. This study was undertaken: 1) to evaluate whether and which indicator(s) of autonomic cardiac regulation and psychophysical stress can identify successful athletes during a training season culminating with the Olympic Games and 2) to evaluate the feasibility of a quasi-on-line assessment of autonomic cardiac regulation from training field, by a telematic approach. METHODS: This study was conducted on the group of male athletes composing the Italian national team of rowing (n=34), in the season preceding the 2004 Olympic Games. Complete results are from 18 subjects (age 25.3+/-0.5 years), who were selected to participate to the Athens' Olympic games. Athletes were studied while partially detrained, at mid-training season and close to the games. The RR interval was obtained through a miniature transtelephonic-ECG recorder in the supine and standing posture, thus allowing the evaluation of cardiovascular responses to a sympathetic challenge. Data were downloaded through a telephone line, to the referral center where RR-interval variability data were analyzed with the autoregressive method. Also, in each study sessions, athletes filled a self-administered questionnaire of stress perception and somatic symptoms (4S-Q). RESULTS: All ECG recordings were transmitted successfully by phone to the referral center. No significant difference was detected in any marker of autonomic cardiac regulation between athletes who won a medal at the Olympic Games and those who did not. However, respiratory rate was faster in medal winners (P=0.02), while the questionnaire addressing stress (4S-Q) provided greater scores in the group that did not win a medal (F=5.55, P<0.022) at mid-training season and close to the Olympic Games. CONCLUSIONS: The results of this study would suggest the possibility of an early detection of psychosomatic symptoms resulting from long duration and elevated stress of preparing for top level competitions, whose better handling might identify the most successful athletes. In addition, it indicates the feasibility of a quasi-on-line assessment of autonomic cardiac adaptations to strenuous training directly from field to be possibly used for improving individual training programs, allowing athletes evaluation in their natural environment. PMID- 17119527 TI - Time course of blood pressure changes immediately after maximal exercise. AB - AIM: The aim of this study was to investigate the effect of exhaustive exercise on the time course of arterial blood pressure (BP) and heart rate (HR) during upright resting (inactive) and loadless pedaling (active) recovery from a bicycle exercise to exhaustion. METHODS: The subjects were 11 healthy normotensive males. Systolic, diastolic and mean BP, and HR were recorded every 20 s for the initial 6 min of the recovery period. RESULTS: The time course of all BP measures during inactive and active recovery was characterized by a marked and sudden drop during the initial 20-s period, followed by a quick rise. This was followed by a gradual decline till the end of the recovery period. The time course of HR recovery, on the other hand, exhibited a smooth decline without the initial drop. With active recovery, the initial drop of diastolic and mean BP was less than the inactive recovery. After the 20 s period, the diastolic BP and HR were kept slightly higher with the active recovery than the inactive recovery. CONCLUSIONS: A sudden drop of the BP occurred at the initial recovery period of postcycle exercise to exhaustion though HR did not show such a change. The initial BP drop could be attenuated by the actively pedaling the cycle without load. PMID- 17119528 TI - Free testosterone/cortisol ratio in soccer: usefulness of a categorization of values. AB - AIM: The free testosterone:cortisol ratio (FTCR) is widely used for studying and preventing overtraining syndrome in various sports. The use of FTCR for following overtraining syndrome was proposed originally with two approaches: FTCR lower than 0.35x10(-3), calculated on free testosterone (FT) in nanomoles per liter (nmol/L) and on cortisol (C) in micromoles per liter (mmole/L) or a decrease of the ratio of 30% or more in comparison with the previous value. In our experience, the use of an absolute value as a threshold is not useful, whereas the evaluation of the concentrations of hormones and their ratio in comparison with previous ones is more useful. These classical approaches are not, however, sufficient to describe the various possible physiological modifications linked to training excess and/or incomplete recovery. METHODS: We collected samples from 32 professional soccer players of an Italian First Division team, during the period July 2001-July 2003. We analyzed the values of 21 athletes during the season 2001 2002 and of 11 athletes during the season 2002-2003 (6 out of 11 were examined also during the previous one) always present when the 4 (first season) or 5 (second season) blood drawings have been performed. We applied an original, pragmatic and easy-to-use classification of FTCR values, in association with classical interpretations based on decreases of the values in comparison with previous athlete's result. RESULTS: We used the traditional approaches in two consecutive seasons in a professional soccer team: the evaluation of the decrease >30% of the parameter in comparison with the previous value or with the basal (preseason) value are shown. The statistical differences between the FTCR values of the six athletes followed in both seasons were not significant. CONCLUSIONS: The classification method we propose is advantageous in comparison with traditional interpretative schemes, because identify different risk categories, stratifying the interval between the values 0.35-0.8. PMID- 17119529 TI - Acute effects of active warm-up and stretching on the flexibility of elderly women. AB - AIM: The purpose of the investigation was to examine the acute effects of static stretch on the range of motion (ROM) of the lower extremities and trunk in elderly women, when stretching is performed with and without warming-up exercises. METHODS: Twenty-two sedentary subjects 65-85 years old (mean age: 76.5 years) with normal ROM without joint abnormalities took part in the study, and performed 3 different flexibility protocols in non-consecutive randomized sessions. The first stretching protocol comprised of a general warming-up for 20 min, the second of the same general warming-up followed by static stretching of the lower extremities and the trunk, whereas the third and final stretching protocol consisted of static stretching alone. Passive ROM was examined at the lower extremity joints and trunk flexion, using a goniometer and a flexometer. RESULTS: Static stretching alone and static stretching after a general warming-up bout, significantly increased the range of all lower extremity joints and trunk flexion (P<0.001). The general warming-up session included a significantly increased ROM only at the ankle dorsiflexion joint (P<0.001). CONCLUSIONS: The results reflect immediate changes in flexibility via acute stretching exercises, in sedentary elderly women, when muscles undergo static elongation, irrespective of the performance of warming-up exercises. PMID- 17119530 TI - The effect of exercise on urinary gamma-glutamyl transferase and protein levels of volleyball players. AB - AIM: Postexercise proteinuria and increased urinary gamma-glutamyl transferase (GGT) levels can be indicative of exercise-induced renal damage. In the literature, there exists numerous studies on exercise-induced proteinuria; but studies investigating the effects of exercise on urinary GGT levels are quite few. We aimed to evaluate the effects of exercise on renal function, expressed through the exercise-induced differences in urinary GGT, creatinine and protein levels. METHODS: The study was performed on 12 female and 12 male volleyball players of the same sports club. Urine samples collected before and 1 h after the exercise were analyzed for urinary GGT, creatinine and protein amounts. RESULTS: No statistically significant difference was observed between pre- and postexercise urinary GGT levels (U/L and U/g creatinine) of female and male volleyball players (P>0.05). A significant exercise-induced increase in urinary protein excretion was observed for the male players, while a significant exercise induced increase in urinary creatinine excretion was observed for the female players (P<0.05). When urinary GGT levels (U/L) were compared separately for setters and spikers, it was observed that female players had no significant difference, while male spikers had a statistically significant exercise-induced increase in the urinary GGT levels (U/L) (P<0.05). CONCLUSIONS: We suggest that the insignificance of the exercise-induced increases in the urinary parameters could be due to the relatively short-course of the exercise and the timing of postexercise urine collection. A comprehensive study performed on more subjects could yield results that are more significant. PMID- 17119531 TI - Toxicology testing and results for suicide victims--13 states, 2004. AB - In 2003, an estimated 31,484 suicides (10.7 per 100,000 population) occurred in the United States. Suicide was the fourth leading cause of death among persons aged 10-64 years and the second and third leading causes of death among persons aged 25-34 and 10-24 years, respectively. Few studies have attempted to determine the contribution of substance use to suicide. To assess toxicology testing practices and to determine the prevalence of positive results for alcohol or other drugs, CDC analyzed test results of suicide victims in the 13 states that collected data for the National Violent Death Reporting System (NVDRS) in 2004. This report summarizes the results of that analysis, which determined that 1) the percentage of suicide victims tested varied among states, ranging from 25.9% to 97.7%; 2) of those victims tested, 33.3% were positive for alcohol, and 16.4% were positive for opiates; and 3) similar percentages of poisoning suicide (i.e., suspected intentional overdose) and nonpoisoning suicide victims tested positive for alcohol or other drugs, with the exception of opiates. These results underscore the need to continue monitoring toxicology test results of suicide victims, which might identify patterns of substance use that can help guide development of effective suicide interventions. Such data can be enhanced by uniform, comprehensive, toxicology testing practices on a state and national basis. PMID- 17119532 TI - Improvement in lipid and glycated hemoglobin control among black adults with diabetes--Raleigh and Greensboro, North Carolina, 1997-2004. AB - Previous studies have indicated that, in the United States, black persons with diabetes have lower levels of glycemic and lipid control and are at increased risk for diabetes-related complications than white persons with the disease. Clinical trials have demonstrated that glycemic and lipid control can reduce the risk for microvascular and macrovascular complications among adults. In addition, recent studies of national survey data have indicated a secular trend of gradual improvements in blood pressure, cholesterol levels, and smoking rates among U.S. persons with diabetes. These studies have demonstrated an increase in the proportion of persons who meet recommended levels for blood pressure, glycated hemoglobin (HbA1c), and cholesterol. Whether black persons in the United States have benefited from these overall improvements is unclear. Surveys conducted among black adults in Raleigh and Greensboro, North Carolina, as part of Project DIRECT (Diabetes Intervention Reaching and Educating Communities Together), provided an opportunity to examine trends in diabetes control and risk for complications. Project DIRECT is a community-based intervention aimed at improving self-care, access to care, and quality of care for residents with diabetes. The analyses described in this report examined whether glycemic and lipid control improved in both communities from 1997 to 2004, a period of rapid advances in clinical understanding of how to control diabetes and its complications. This report summarizes the results of those analyses, which indicated improvements in the proportion of black adults with diabetes who reported that they were meeting recommended levels of HbA1c, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol, triglycerides, and total cholesterol. However, a substantial number of these persons smoked and were above recommended glycemic and lipid levels at follow-up. Therefore, continued education of the public is important in improving quality of care and reducing risk factors for persons at high risk for diabetes and cardiovascular disease. PMID- 17119533 TI - Geographic disparities in diabetes-related amputations--Texas-Mexico border, 2003. AB - The risk for lower extremity amputation (LEA) is estimated at 15 to 40 times higher among persons with diabetes than among persons without diabetes. In Texas, the prevalence of diabetes is higher near the Mexico border, where persons are more likely to have lower levels of education, lower incomes, no health insurance, and other barriers to obtaining health care. To determine whether diabetes-related LEA rates are higher near the Texas-Mexico border, rates were calculated, in both the general population and among persons with diabetes, for diabetes-related LEAs in border and nonborder counties. Data used for this analysis included 2003 Texas Inpatient Hospital Discharge Data (TIHDD), 2003 Texas population estimates, and data from the 2003 Texas Behavioral Risk Factor Surveillance System (BRFSS). The results of the analysis indicated that the age- and sex-adjusted rate of diabetes-related LEAs in the general population along the border was nearly double the rate of nonborder counties. Among persons with diabetes, the rate along the border also was significantly higher than among those in nonborder counties, but the rate differences were primarily among men aged > or =45 years. Additional measures to prevent diabetes and improve education regarding diabetes care are needed to reduce the excess burden of LEAs among persons with diabetes along the border. PMID- 17119534 TI - Abortion surveillance--United States, 2003. AB - PROBLEM/CONDITION: CDC began abortion surveillance in 1969 to document the number and characteristics of women obtaining legal induced abortions. REPORTING PERIOD COVERED: This report summarizes and describes data voluntarily reported to CDC regarding legal induced abortions obtained in the United States in 2003. DESCRIPTION OF SYSTEM: For each year since 1969, CDC has compiled abortion data by state or area of occurrence. During 1973-1997, data were received from or estimated for 52 reporting areas in the United States: 50 states, the District of Columbia, and New York City. In 1998 and 1999, CDC compiled abortion data from 48 reporting areas. Alaska, California, New Hampshire, and Oklahoma did not report, and data for these states were not estimated. During 2000-2002, Oklahoma again reported these data, increasing the number of reporting areas to 49, and for 2003, Alaska again reported and West Virginia did not, maintaining the number of reporting areas at 49. RESULTS: A total of 848,163 legal induced abortions were reported to CDC for 2003 from 49 reporting areas, representing a 0.7% decline from the 854,122 legal induced abortions reported by 49 reporting areas for 2002. The abortion ratio, defined as the number of abortions per 1,000 live births, was 241 in 2003, a decrease from the 246 in 2002. The abortion rate was 16 per 1,000 women aged 15-44 years for 2003, the same as for 2002. For the same 47 reporting areas, the abortion rate remained relatively constant during 1998-2003. During 2001-2002 (the most recent years for which data are available), 15 women died as a result of complications from known legal induced abortion. One death was associated with known illegal abortion. The highest percentages of reported abortions were for women who were unmarried (82%), white (55%), and aged <25 years (51%). Of all abortions for which gestational age was reported, 61% were performed at < or =8 weeks' gestation and 88% at <13 weeks. From 1992 (when detailed data regarding early abortions were first collected) through 2002, steady increases have occurred in the percentage of abortions performed at < or =6 weeks' gestation, with a slight decline in 2003. A limited number of abortions were obtained at >15 weeks' gestation, including 4.2% at 16--20 weeks and 1.4% at > or =21 weeks. A total of 36 reporting areas submitted data documenting that they performed and enumerated medical (nonsurgical) procedures, making up 8.0% of all known reported procedures from the 45 areas with adequate reporting on type of procedure. INTERPRETATION: During 1990-1997, the number of legal induced abortions gradually declined. When the same 47 reporting areas are compared, the number of abortions decreased during 1996-2001, then slightly increased in 2002 and again decreased in 2003. In 2000 and 2001, even with one additional reporting state, the number of abortions declined slightly, with a minimal increase in 2002 and a further decrease in 2003. In 2001 and 2002, as in the previous years, deaths related to legal induced abortions occurred rarely. PUBLIC HEALTH ACTION: Abortion surveillance in the United States continues to provide the data necessary for examining trends in numbers and characteristics of women who obtain legal induced abortions and to increase understanding of this pregnancy outcome. Policymakers and program planners use these data to improve the health and well being of women and infants. PMID- 17119535 TI - Replication of reported linkages for dyslexia and spelling and suggestive evidence for novel regions on chromosomes 4 and 17. AB - We report the first genome-wide linkage analysis for reading and spelling in a sample of 403 families of twins, aged between 12 and 25 years taken from the normal population and unselected for reading ability. These traits showed heritabilities of 0.52-0.73, and support for linkage exceeded replication levels (lod > 1.44) of seven of the 11 linkages reported in dyslexic samples, namely: 2q22.3, 3p12-q13, 6q11.2, 7q32, 15q21.1, 18p21, and Xq27.3. For five of these (2q22.3, 6q11.2, 7q32, 18p21, and Xq27), this study provides the first independent replication. 1p34-36 and 2p15-16 received some support, with lods of 1.2 and 0.83, respectively, whereas two regions received little support (6p23 21.3 and 11p15.5). This study also identified two novel linkages at 4p15.33-16.1 and 17p13.3, which received suggestive support (max. lod 2.08 and 1.99, respectively). PMID- 17119536 TI - Detection of small genomic imbalances using microarray-based multiplex amplifiable probe hybridization. AB - Array-based genome-wide screening methods were recently introduced to clinical practice in order to detect small genomic imbalances that may cause severe genetic disorders. The continuous advancement of such methods plays an extremely important role in diagnostic genetics and medical genomics. We have modified and adapted the original multiplex amplifiable probe hybridization (MAPH) to a novel microarray format providing an important new diagnostic tool for detection of small size copy-number changes in any locus of human genome. Here, we describe the new array-MAPH diagnostic method and show proof of concept through fabrication, interrogation and validation of a human chromosome X-specific array. We have developed new bioinformatic tools and methodology for designing and producing amplifiable hybridization probes (200-600 bp) for array-MAPH. We designed 558 chromosome X-specific probes with median spacing 238 kb and 107 autosomal probes, which were spotted onto microarrays. DNA samples from normal individuals and patients with known and unknown chromosome X aberrations were analyzed for validation. Array-MAPH detected exactly the same deletions and duplications in blind studies, as well as other unknown small size deletions showing its accuracy and sensitivity. All results were confirmed by fluorescence in situ hybridization and probe-specific PCR. Array-MAPH is a new microarray based diagnostic tool for the detection of small-scale copy-number changes in complex genomes, which may be useful for genotype-phenotype correlations, identification of new genes, studying genetic variation and provision of genetic services. PMID- 17119537 TI - Preclinical investigation of the functional effects of memantine and memantine combined with galantamine or donepezil. AB - Combinations of drugs approved to treat Alzheimer's disease (AD) were tested in older rabbits with delay eyeblink classical conditioning, a form of associative learning severely impaired in AD. In Experiment 1 (n=49 rabbits), low doses (0.1, 0.5, 1.0, and 0.0 (vehicle) mg/kg) of memantine (Namenda) were tested. These three doses neither improved nor impaired acquisition at a statistically significant level. The 0.5 mg/kg dose had the greatest effect numerically and did not cause sensitization or habituation in explicitly unpaired controls. In Experiment 2 (n=56), doses of galantamine (Razadyne; 3.0 mg/kg) and donepezil (Aricept; 0.75 mg/kg) that had comparable magnitudes of cholinesterase inhibition were tested alone and in combination with 0.5 mg/kg memantine. Older rabbits treated with galantamine and with galantamine+memantine learned significantly better than vehicle-treated rabbits, but adding memantine did not improve learning over galantamine alone. Older rabbits treated with donepezil or a combination of memantine and donepezil did not learn significantly better than rabbits treated with vehicle. Galantamine has two mechanisms of action: mild cholinesterase inhibition and allosteric modulation of nicotinic acetylcholine receptors (nAChRs). When equated for cholinesterase inhibition, galantamine had significant efficacy in the eyeblink conditioning model system, but donepezil did not, indicating that modulation of nAChRs may be the mechanism that significantly ameliorates learning deficits in this model. In the absence of AD neuropathology in older rabbits, memantine had no efficacy alone or in combination with the other drugs. PMID- 17119538 TI - Acute effects of the ampakine farampator on memory and information processing in healthy elderly volunteers. AB - Ampakines act as positive allosteric modulators of AMPA-type glutamate receptors and facilitate hippocampal long-term potentiation (LTP), a mechanism associated with memory storage and consolidation. The present study investigated the acute effects of farampator, 1-(benzofurazan-5-ylcarbonyl) piperidine, on memory and information processes in healthy elderly volunteers. A double-blind, placebo controlled, randomized, cross-over study was performed in 16 healthy, elderly volunteers (eight male, eight female; mean age 66.1, SD 4.5 years). All subjects received farampator (500 mg) and placebo. Testing took place 1 h after drug intake, which was around Tmax for farampator. Subjects performed tasks assessing episodic memory (wordlist learning and picture memory), working and short-term memory (N-back, symbol recall) and motor learning (maze task, pursuit rotor). Information processing was assessed with a tangled lines task, the symbol digit substitution test (SDST) and the continuous trail making test (CTMT). Farampator (500 mg) unequivocally improved short-term memory but appeared to impair episodic memory. Furthermore, it tended to decrease the number of switching errors in the CTMT. Drug-induced side effects (SEs) included headache, somnolence and nausea. Subjects with SEs had significantly higher plasma levels of farampator than subjects without SEs. Additional analyses revealed that in the farampator condition the group without SEs showed a significantly superior memory performance relative to the group with SEs. The positive results on short-term memory and the favorable trends in the trail making test (CTMT) are interesting in view of the development of ampakines in the treatment of Alzheimer's disease and schizophrenia. PMID- 17119539 TI - 2-Aminotetraline derivative protects from ischemia/reperfusion brain injury with a broad therapeutic window. AB - The effect of ST1942, a 2-aminotetraline derivative with anti-inflammatory properties, was evaluated in ischemia/reperfusion injury in CD1 and C57BL/6 mice. ST1942 or saline were injected intraperitoneally 30 min and 6, 24, 36 h after ischemia. Forty-eight hours after ischemia, ST1942 (25 mg/kg) reduced the infarct volume by 50% in CD1 and 61% in C57BL/6 mice. All subsequent data were obtained from the latter strain. The ischemic lesion was significantly reduced by 30% when the first injection was administered 6 h after ischemia, revealing a broad effective window. Degenerating neurons in striatum, cortex and hippocampus of ischemic mice were markedly decreased by ST1942. Also examined was the effect of ST1942 on general and focal neurological deficits for 4 days after ischemia. Mice receiving the drug twice daily showed constantly reduced deficits. We then investigated the cortical mRNA expression of some inflammatory and apoptotic genes by real-time PCR. Forty-eight hours after ischemia ST1942 treatment significantly counteracted ischemia-induced activation of IL-1beta, TNFalpha, and Bax, and enhanced the expression of the antiapoptotic gene, Bcl-2, showing in vivo anti-inflammatory and antiapoptotic actions. The microglial activation/macrophage recruitment in the ischemic lesion was strongly prevented in mice receiving ST1942. In neuron-microglia cocultures, ST1942 significantly counteracted LPS-induced cytotoxicity. Binding data and experiments on microglial cell cultures indicate that the anti-inflammatory effect of ST1942 may be due to its action on 5-HT2B receptors, thus highlighting the possibility that this 5-HT receptor subtype may represent a novel target for neuroprotective drugs in ischemic injury. PMID- 17119540 TI - Chronic fluoxetine treatment attenuates stressor-induced changes in temperature, heart rate, and neuronal activation in the olfactory bulbectomized rat. AB - The olfactory bulbectomized (OB) rat is a well-characterized animal model that exhibits a number of behavioral and neurochemical changes that have relevance to clinical depression. Hyperactivity in the open field is the most widely used parameter assessed in this model and is reversed following chronic, but not acute, antidepressant treatment. This study investigated OB-induced alterations in heart rate, body temperature, and neuronal activation following open-field exposure and the impact of chronic treatment with fluoxetine on these parameters. Upon placement in the open field, OB rats exhibited a characteristic hyperactivity response. Heart rate and body temperature were increased in sham operated rats following open-field exposure, a predictable response to stress, which was significantly reduced in OB rats. Moreover bulbectomy reduced open field-induced cFOS expression in the basal nucleus of the stria terminalis while concurrently increasing expression in the hippocampus, amygdala, paraventricular nucleus of the thalamus, and dorsal raphe nucleus. Chronic fluoxetine treatment (10 mg/kg subcutaneous once daily for 5 weeks) attenuated all of these OB associated changes. In conclusion, OB rats exhibit alterations in behavior, body temperature, heart rate, and neuronal activation in response to open-field exposure, which are reversed following chronic fluoxetine administration. These results identify stress-sensitive regions within the brain which are altered following bulbectomy and which may underlie the abnormal behavioral and physiological changes observed in this rodent model of depression. PMID- 17119541 TI - Effects of peroxisome proliferator-activated receptor gamma agonists on brain glucose and glutamate transporters after stress in rats. AB - Repeated stress causes an energy-compromised status in the brain, with a decrease in glucose utilization by the brain cells, which might account for excitotoxicity processes seen in this condition. In fact, brain glucose metabolism mechanisms are impaired in some neurodegenerative disorders, including stress-related neuropsychopathologies. More recently, it has been demonstrated that some synthetic peroxisome proliferator-activated receptor gamma (PPARgamma) agonists increase glucose utilization in rat cortical slices and astrocytes, as well as inhibit brain oxidative damage after repeated stress, which add support for considering these drugs as potential neuroprotective agents. To assess if stress causes glucose utilization impairment in the brain and to study the mechanisms by which this effect is achieved, young-adult male Wistar rats (control and immobilized for 6 h during 7 or 14 consecutive days, S7, S14) were i.p. injected with the natural ligand 15-deoxy-Delta-12,14-prostaglandin J2 (PGJ2, 120 microg/kg) or the high-affinity ligand rosiglitazone (RG, 3 mg/kg) at the onset of stress. Repeated immobilization during 1 or 2 weeks produces a decrease in brain cortical synaptosomal glucose uptake, and this effect was prevented by treatment with both natural and synthetic PPARgamma ligands by restoring protein expression of the neuronal glucose transporter, GLUT-3 in membrane fractions. On the other hand, treatment with PPARgamma ligands prevents stress-induced ATP loss in rat brain. Finally, repeated immobilization stress also produces a decrease in brain cortical synaptosomal glutamate uptake, and this effect was prevented by treatment with PPARgamma ligands by restoring synaptosomal protein expression of the glial glutamate transporter, EAAT2. In summary, our results demonstrate that 15d-PGJ2 and the thiazolidinedione rosiglitazone increase neuronal glucose metabolism, restore brain ATP levels and prevent the impairment in glutamate uptake mechanisms induced by exposure to stress, suggesting that this class of drugs may be therapeutically useful in conditions in which brain glucose levels or availability are limited after exposure to stress. PMID- 17119542 TI - Endocannabinoids in chronic migraine: CSF findings suggest a system failure. AB - Based on experimental evidence of the antinociceptive action of endocannabinoids and their role in the modulation of trigeminovascular system activation, we hypothesized that the endocannabinoid system may be dysfunctional in chronic migraine (CM). We examined whether the concentrations of N arachidonoylethanolamide (anandamide, AEA), palmitoylethanolamide (PEA), and 2 arachidonoylglycerol (2-AG) in the CSF of patients with CM and with probable CM and probable analgesic-overuse headache (PCM+PAOH) are altered compared with control subjects. The above endocannabinoids were measured by high-performance liquid chromatography (HPLC), and quantified by isotope dilution gas chromatography/mass-spectrometry. Calcitonin gene-related peptide (CGRP) levels were also determined by RIA method and the end products of nitric oxide (NO), the nitrites, by HPLC. CSF concentrations of AEA were significantly lower and those of PEA slightly but significantly higher both in patients with CM and PCM+PAOH than in nonmigraineur controls (p<0.01 and p<0.02, respectively). A negative correlation was found between AEA and CGRP levels in CM and PCM+PAOH patients (r=0.59, p<0.01 and r=-0.65, p<0.007; respectively). A similar trend was observed between this endocannabinoid and nitrite levels. Reduced levels of AEA in the CSF of CM and PCM+PAOH patients may reflect an impairment of the endocannabinoid system in these patients, which may contribute to chronic head pain and seem to be related to increased CGRP and NO production. These findings support the potential role of the cannabinoid (CB)1 receptor as a possible therapeutic target in CM. PMID- 17119543 TI - STN stimulation alters pallidal-frontal coupling during response selection under competition. AB - To investigate the effects of bilateral subthalamic nucleus (STN) stimulation on patterns of brain activation during random number generation (RNG), a task that requires suppression of habitual counting and response selection under competition. We used H(2)(15)O positron emission tomography to investigate the changes of regional cerebral blood flow (rCBF) induced by bilateral STN stimulation during a RNG task, in six patients with Parkinson's disease. Paced RNG at 1 Hz was compared with a control counting task. Both tasks were performed off medication with deep brain stimulation on and off. Subthalamic nucleus stimulation had a negative effect on performance of fast-paced RNG, leading to reduced randomness and increased habitual counting. Subthalamic nucleus stimulation also induced a reduction of rCBF in the left dorsal frontal gyrus, inferior frontal gyrus, dorsolateral prefrontal cortex, posterior and right anterior cingulate, and an increase of rCBF in the right internal globus pallidum (GPi) during RNG. Stimulation of the STN significantly altered pallidal coupling with frontal and temporal areas compared with when the stimulators were off. In conclusion, during RNG: (i) STN stimulation activates its output neurons to the GPi; (ii) STN stimulation induces increased inhibition of a prefrontal-cingulate network. This is the first direct evidence that STN stimulation significantly alters pallidal coupling with prefrontal, cingulate, and temporal cortices during performance of a task that requires response selection under competition. PMID- 17119544 TI - Neuroprotection in ischemic mouse brain induced by stem cell-derived brain implants. AB - Protective mechanisms of the brain may reduce the extent of injury after focal cerebral ischemia. Here, we explored in a mouse model of focal cerebral ischemia potential synergistic neuroprotective effects of two mediators of neuroprotection: (i) neuronal or glial precursor cells and (ii) the inhibitory neuromodulator adenosine. Embryonic stem (ES) cells, engineered to release adenosine by biallelic disruption of the adenosine kinase gene, and respective wild-type cells were induced to differentiate into either neural or glial precursor cells and were injected into the striatum of mice 1 week before middle cerebral artery occlusion. All stem cell-derived graft recipients were characterized by a significant reduction in infarct volume, an effect that was augmented by the release of adenosine. Neuroprotection was strongest in adenosine releasing glial precursor cell recipients, which were characterized by an 85% reduction of the infarct area. Graft-mediated neuroprotection correlated with a significant improvement of general and focal neurologic scores. Histologic analysis before and after ischemia revealed clusters of implanted cells within the striatum of all treated mice. We conclude that ES cell derived adenosine releasing brain implants provide neuroprotection by synergism of endogenous precursor cell-mediated effects and paracrine adenosine release. PMID- 17119545 TI - Muscularity and adiposity in addition to net acid excretion as predictors of 24-h urinary pH in young adults and elderly. AB - OBJECTIVE: In patients with nephrolithiasis, an inverse relationship between 24-h urinary pH (24h-UpH) and body weight has been reported. Whether body composition indices and 24h-UpH are similarly associated in healthy subjects needs investigation. DESIGN: Cross-sectional, retrospective analysis. SETTING: Dortmund, Germany and Gothenburg, Sweden. SUBJECTS: Healthy young adults (18-23 years; n=117) and elderly (55-75 years; n=85) having a mean body mass index (BMI) of 22.80+/-3.4 and 25.3+/-3.9 kg/m2, respectively. METHODS: Anthropometric data, 24h-UpH, and 24-h urinary excretion rates of net acid (NAE), creatinine, and urea were determined. After adjusting for urea (reflecting protein intake), renal creatinine output was used as a biochemical marker for muscularity. The BMI served as a marker of adiposity. RESULTS: NAE, body weight, and BMI were significantly (P<0.05) higher, and height and creatinine significantly lower in the elderly, whereas body-surface area (BSA) was not different. Step-wise multiple regression analysis using BSA-corrected urinary variables revealed NAE as the primary predictor of 24h-UpH (with R2 values of 0.64 and 0.68 in young adults and elderly, respectively, P<0.0001), followed by urea (P<0.0001), creatinine (P<0.05), and BMI (P<0.05 for the young adults and P=0.12 for the elderly). These associations were negative for NAE and BMI, and positive for urea and creatinine. CONCLUSIONS: Muscularity (i.e. creatinine adjusted for urea) and particularly in the group of young adults, adiposity (i.e. BMI) proved to be modest, but significant predictors of 24h-UpH. Future research should focus on more obese subjects in whom insulin resistance and particular kidney functions should also be examined to further substantiate the role of obesity in low-urine pH-associated conditions, for example, nephrolithiasis. PMID- 17119546 TI - Prevalence of the metabolic syndrome among Turkish adults. AB - OBJECTIVE: To determine prevalence of the metabolic syndrome (MS) in a sample representing Turkish population using United States Adult Treatment Panel-3 guidelines. DESIGN: The study included random samples from both urban and rural populations in the seven geographical regions of Turkey. The population for this analysis were 2108 men (1372 in urban and 736 in rural areas) and 2151 women (1423 in urban and 728 in rural areas) with a mean age of 40.9+/-14.9 years (range 20-90). RESULTS: The prevalence of the MS diagnosed using the Adult Treatment Panel III criteria was 33.9% (1442 of 4259) and differed significantly in men (28%) and women (39.6%). The prevalence of syndrome increased with age in men, from 10.7% in subjects aged 20-29 years to 49% in those aged over 70 years. The prevalence increased with age in women, from 9.6% in subjects aged 20-29 years to 74.6% in those aged 60-69 years, and decreased to 68.6% in those over 70 years of age. The prevalence of the syndrome was similar in urban (33.8%) and rural (33.9%) population. We found 26.8, 26.4, 19.3, 10.9 and 3.6% of the population had at least 1, 2, 3, 4 or 5 components, respectively. We found 57.2, 32.3 and 10.6% of the subjects with MS had 3, 4 and 5 components, respectively. CONCLUSIONS: The prevalence of the MS in the adult Turkish population is very high, especially in women. Our findings have important implications for public health in Turkey. PMID- 17119547 TI - A method to improve the nutritional quality of foods and beverages based on dietary recommendations. AB - OBJECTIVE: The increasing consumer interest in health prompted Unilever to develop a globally applicable method (Nutrition Score) to evaluate and improve the nutritional composition of its foods and beverages portfolio. METHODS: Based on (inter)national dietary recommendations, generic benchmarks were developed to evaluate foods and beverages on their content of trans fatty acids, saturated fatty acids, sodium and sugars. High intakes of these key nutrients are associated with undesirable health effects. In principle, the developed generic benchmarks can be applied globally for any food and beverage product. Product category-specific benchmarks were developed when it was not feasible to meet generic benchmarks because of technological and/or taste factors. RESULTS: The whole Unilever global foods and beverages portfolio has been evaluated and actions have been taken to improve the nutritional quality. The advantages of this method over other initiatives to assess the nutritional quality of foods are that it is based on the latest nutritional scientific insights and its global applicability. CONCLUSIONS: The Nutrition Score is the first simple, transparent and straightforward method that can be applied globally and across all food and beverage categories to evaluate the nutritional composition. It can help food manufacturers to improve the nutritional value of their products. In addition, the Nutrition Score can be a starting point for a powerful health indicator front of-pack. This can have a significant positive impact on public health, especially when implemented by all food manufacturers. PMID- 17119550 TI - Sexual selection and the evolution of evolvability. AB - Here we show that sexual selection can have an effect on the rate of mutation. We simulated the fate of a genetic modifier of the mutation rate in a sexual population with and without sexual selection (modelled using a female choice mechanism). Female choice for 'good genes' should reduce variability among male subjects, leaving insufficient differences to maintain female preferences. However, female choice can actually increase genetic variability by supporting a higher mutation rate in sexually selected traits. Increasing the mutation rate will be selected against because of the resulting decline in mean fitness. However, it also increases the probability of rare beneficial mutations arising, and mating skew caused by female preferences for male subjects carrying those beneficials with few deleterious mutations ('good genes') can lead to a mutation rate above that expected under natural selection. A choice of two male subjects was sufficient for there to be a twofold increase in the mutation rate as opposed to a decrease found under random mating. PMID- 17119551 TI - Individual QTLs controlling quantitative variation in blood pressure inherited in a Mendelian mode. AB - We studied three possible genotypes at 10 well-defined blood pressure (BP) QTLs using congenic rat lines. The central question was whether the hypertensive or normotensive allele is dominant, or whether there is partial dominance. The congenic strains were employed to investigate the BP effects of alleles originating from normotensive rats in the background of hypertensive Dahl salt sensitive (DSS) rats. The normotensive alleles at eight QTLs were fully dominant over DSS alleles, which we tentatively interpreted as indicating that DSS rats incurred a loss of function at these loci and that the QTLs produced BP-reducing agents. In contrast, the normotensive allele of only one QTL was recessive over its DSS counterpart, implying a gain of function at this QTL or a null allele involved in generating a BP-elevating agent. Only one locus, C17QTL, had alleles exhibiting partial dominance. These estimates of dominance differ considerably from those obtained by QTL analysis in a F2 cross. This disagreement demonstrates the importance of establishing a cause-effect relationship between a QTL and its phenotypic effect via congenic strains. The dominance relationships suggest pertinent strategies for gene identification and pharmaceutical intervention. PMID- 17119552 TI - Time-delay compensated monochromator in the off-plane mount for extreme ultraviolet ultrashort pulses. AB - The design of ultrafast monochromators using grazing-incidence gratings in the off-plane mount for the spectral selection of extreme-ultraviolet femtosecond pulses in a broad spectral region is presented. Their application in the selection of high-order laser harmonics is analyzed in detail. The main advantage of the off-plane mount is a much higher efficiency than that of the classical mount. It is shown that two-grating configurations preserve the length of the optical paths of different diffracted rays, maintaining the extremely short time duration of the pulse. Configurations with plane or toroidal gratings are discussed. As a test case, the design of a monochromator for the 17-61 nm region with a time compensation better than 1 fs is presented. PMID- 17119553 TI - Analysis of optical systems with extended depth of field using the Wigner distribution function. AB - We describe the use of a Wigner distribution function approach for exploring the problem of extending the depth of field in a hybrid imaging system. The Wigner distribution function, in connection with the phase-space curve that formulates a joint phase-space description of an optical field, is employed as a tool to display and characterize the evolving behavior of the amplitude point spread function as a wave propagating along the optical axis. It provides a comprehensive exhibition of the characteristics for the hybrid imaging system in extending the depth of field from both wave optics and geometrical optics. We use it to analyze several well-known optical designs in extending the depth of field from a new viewpoint. The relationships between this approach and the earlier ambiguity function approach are also briefly investigated. PMID- 17119554 TI - Complete wavefront reconstruction using sequential intensity measurements of a volume speckle field. AB - The recording of the volume speckle field from an object at different planes combined with the wave propagation equation allows the reconstruction of the wavefront phase and amplitude without requiring a reference wave. The main advantage of this single-beam multiple-intensity reconstruction (SBMIR) technique is the simple experimental setup because no reference wave is required as in the case of holography. The phase retrieval technique is applied to the investigation of diffusely transmitting and reflecting objects. The effects of different parameters on the quality of reconstructions are investigated by simulation and experiment. Significant enhancements of the reconstructions are observed when the number of intensity measurements is 15 or more and the sequential measurement distance is 0.5 mm or larger. Performing two iterations during the reconstruction process using the calculated phase also leads to better reconstruction. The results from computer simulations confirm the experiments. Analysis of transverse and longitudinal intensity distributions of a volume speckle field for the SBMIR technique is presented. Enhancing the resolution method by shifting the camera a distance of a half-pixel in the lateral direction improves the sampling of speckle patterns and leads to better quality reconstructions. This allows the possibility of recording wave fields from larger test objects. PMID- 17119555 TI - Quasi-absolute measurement of aspheres with a combined diffractive optical element as reference. AB - We have already reported a method for the quasi-absolute test of rotationally symmetric aspheres by means of combined diffractive optical elements (combo DOEs). The combo-DOEs carry the information for the ideal shape of an aspheric surface under test as well as a spherical wave for the measurement at the cat's eye position. An experimental demonstration of the procedure is given. Measurements with two different designs of combo-DOEs have been conducted, and their relative advantages and disadvantages are discussed. PMID- 17119556 TI - Electronically swept millimeter-wave interferometer for spatially resolved measurement of plasma electron density. AB - We report the development and initial implementation of what we believe to be a new rapid- spatial-scan millimeter-wave interferometer for plasma density measurements. The fast scan is effected by electronic frequency sweeping of a wideband (180-280 GHz) backward-wave oscillator whose output is focused onto a fixed blazed diffraction grating. The system, which augments the rotating-grating scanned multiview H-1 heliac interferometer, can sweep the plasma cross section in a period of less than 1 ms with a beam diameter in the plasma of 20 mm and phase noise of the order of 0.01 rad. PMID- 17119557 TI - Modeling the interferometric radius measurement using Gaussian beam propagation. AB - We model the interferometric radius measurement using Gaussian beam propagation to identify biases in the measurement due to using a simple geometric ray-trace model instead of the more complex Gaussian model. The radius measurement is based on using an interferometer to identify the test part's position when it is at two null locations, and the distance between the positions is an estimate of the part's radius. The null condition is observed when there is no difference in curvature between the reflected reference and the test wavefronts, and a Gaussian model will provide a first-order estimate of curvature changes due to wave propagation and therefore changes to the radius measurement. We show that the geometric ray assumption leads to radius biases (errors) that are a strong function of the test part radius and increase as the radius of the part decreases. We tested for a bias for both microscaled (<1 mm) and macroscaled parts. The bias is of the order of parts in 10(5) for micro-optics with radii a small fraction of a millimeter and much smaller for macroscaled optics. The amount of bias depends on the interferometer configuration (numerical aperture, etc.), the nominal radius of the test part, and the distances in the interferometer. PMID- 17119558 TI - Effective wavelength calibration for moire fringe projection. AB - The fringe patterns seen when using moire instruments are similar to the patterns seen in traditional interferometry but differ in the spacing between consecutive fringes. In traditional interferometry, the spacing is constant and related to the wavelength of the source. In moire fringe projection, the spacing (the effective wavelength) may not be constant over the field of view and the spacing depends on the system geometry. In these cases, using a constant effective wavelength over the field of view causes inaccurate surface height measurements. We examine the calibration process of the moire fringe projection measurement, which takes this varying wavelength into account to produce a pixel-by-pixel wavelength map. The wavelength calibration procedure is to move the object in the out-of-plane direction a known distance until every pixel intensity value goes through at least one cycle. A sinusoidal function is then fit to the data to extract the effective wavelength pixel by pixel, yielding an effective wavelength map. A calibrated step height was used to validate the effective wavelength map with results within 1% of the nominal value of the step height. The error sources that contributed to the uncertainty in determining the height of the artifact are also investigated. PMID- 17119559 TI - Spectrally resolved phase-shifting interferometry of transparent thin films: sensitivity of thickness measurements. AB - Spectrally resolved white-light phase-shifting interference microscopy can be used for rapid and accurate measurements of the thickness profile of transparent thin-film layers deposited upon patterned structures exhibiting steps and discontinuities. We examine the sensitivity of this technique and show that it depends on the thickness of the thin-film layer as well as its refractive index. The results of this analysis are also valid for any other method based on measurements of the spectral phase such as wavelength scanning or white-light interferometry. PMID- 17119560 TI - Complex amplitude correlation for compensation of large in-plane motion in digital speckle pattern interferometry. AB - The use of complex amplitude correlation to compensate for large in-plane motion in digital speckle pattern interferometry is investigated. The result is compared with experiments where digital speckle photography (DSP) is used for compensation. An advantage of using complex amplitude correlation instead of intensity correlation (as in DSP) is that the phase change describing the deformation is retrieved directly from the correlation peak, and there is no need to compensate for the large movement and then use the interferometric algorithms. A discovered drawback of this method is that the correlation values drop quickly if a phase gradient larger than pi is present in the subimages used for cross correlation. This means that, for the complex amplitude correlation to be used, the size of the subimages must be well chosen or a third parameter in the cross correlation algorithm that compensates for the phase variation is needed. Correlation values and wrapped phase maps from the two techniques (intensity and complex amplitude correlation) are presented. PMID- 17119561 TI - Effect of polarization state on electro-optic coupling and its application to polarization rotation. AB - The effect of the polarization state on electro-optic coupling is studied by using the wave coupling theory of the linear electro-optic effect. The numerical results show that the polarization state obviously influences the electro-optic coupling. The conditions for realizing perfect coupling are emphasized. As an application of perfect coupling, a novel polarization rotator, which can rotate the polarization of a light beam with an arbitrary angle but keep the output intensity unchanged, is presented. PMID- 17119562 TI - Optical data storage system with a planoellipsoidal solid immersion mirror illuminated directly by a point light source. AB - A new solid immersion mirror called the planoellipsoidal (PE) solid immersion mirror (SIM) for the near-field optical storage is proposed and developed. The PE SIM has a small aperture on the apex of the ellipsoidal surface. The intensity distribution of the transmitted field is calculated by using the vector diffraction theory. Compared with a conventional solid immersion lens (SIL), the proposed PE SIM has the following features. A PE SIM replaces three optical elements of the collimator, objective, and SIL in a conventional SIL optical storage system, so that the optical system equipped with the PE SIM is not only simple in its assembly but is also effective in making an optical head unit. The PE SIM obtains light from a point light source and focuses it directly on the recording layer, which may be useful for a compact optical data storage system. The convex ellipsoidal surface of the PE SIM can reduce the risk of the SIM touching the surface of the recording medium. In addition, the spreading of the spot size with the increase of distance is very small in the PE SIM. PMID- 17119563 TI - Wide-angle catadioptric lens with a rectilinear projection scheme. AB - A catadioptric wide-angle lens having a rectilinear projection scheme has been developed with a view to possible applications in the security-surveillance area. The lens has been designed for a miniature camera with a video graphics array grade 1/3 in. color CCD sensor. The field of view of the lens is over 151 degrees , and still distortion is under 1%. Furthermore, the modulation transfer function is better than 0.3 at 70 line pairs/mm over the whole active area of the image sensor. PMID- 17119564 TI - Optics and mechanisms for the Extreme-Ultraviolet Imaging Spectrometer on the Solar-B satellite. AB - The Extreme-Ultraviolet Imaging Spectrometer (EIS) is the first of a new generation of normal-incidence, two-optical-element spectroscopic instruments developed for space solar extreme-ultraviolet astronomy. The instrument is currently mounted on the Solar-B satellite for a planned launch in late 2006. The instrument observes in two spectral bands, 170-210 A and 250-290 A. The spectrograph geometry and grating prescription were optimized to obtain excellent imaging while still maintaining readily achievable physical and fabrication tolerances. A refined technique using low ruling density surrogate gratings and optical metrology was developed to align the instrument with visible light. Slit rasters of the solar surface are obtained by mechanically tilting the mirror. A slit exchange mechanism allows selection among four slits at the telescope focal plane. Each slit is precisely located at the focal plane. The spectrograph imaging performance was optically characterized in the laboratory. The resolution was measured using the Mg iii and Ne iii lines in the range of 171-200 A. The He ii line at 256 A and Ne iii lines were used in the range of 251-284 A. The measurements demonstrate an equivalent resolution of ~2 arc sec? on the solar surface, in good agreement with the predicted performance. We describe the EIS optics, mechanisms, and measured performance. PMID- 17119565 TI - Laboratory calibration of the Extreme-Ultraviolet Imaging Spectrometer for the Solar-B satellite. AB - The laboratory end-to-end testing of the Extreme-Ultraviolet Imaging Spectrometer (EIS) for the Solar-B satellite is reported. A short overview of the EIS, which observes in two bands in the extreme-ultraviolet wavelength range, is given. The calibration apparatus is described, including details of the light sources used. The data reduction and analysis procedure are outlined. The wavelength calibration using a Penning source to illuminate the aperture fully is presented. We discuss the aperture determination using a radiometrically calibrated hollow cathode-based source. We then give an account of the predicted and measured efficiencies from consideration of the efficiencies of individual optical elements in first order, an account of efficiencies out of band when radiation incident in one band is detected in the other, and efficiencies in multiple orders. The efficiencies measured in first order for in band and out of band are compared with the predictions and the sensitivity, and its uncertainties are derived. Application of the radiometric calibration is discussed. PMID- 17119566 TI - Anamorphic white light Fourier processor with holographic lenses. AB - Two anamorphic and achromatic Fourier processors were designed and constructed using diffractive and refractive cylindrical lenses. The diffractive lenses are holographic lenses recorded on silver halide material. In both processors the achromatic one-dimensional Fourier transform plane was obtained with two holographic lenses and one refractive cylindrical lens. The image with the same magnification in both directions at the output plane was formed with two different combinations of lenses. The differences between the two processors are analyzed, and in both cases the chromatic aberration in the Fourier plane and in the output plane is evaluated. Even though single cylindrical refractive lenses were used to image in one direction, good results were obtained. PMID- 17119567 TI - Design of cascaded volume holographic gratings to increase the number of channels for an optical demultiplexer. AB - The design and demonstration of a holographic optical demultiplexer based on cascaded volume holographic gratings are presented. By serially adding a second holographic grating, which has a different grating period, slant angle, and center wavelength compared with those of the first grating, the operating wavelength range of the optical demultiplexer could be expanded, and, therefore, the number of channels of the holographic demultiplexer is doubled. As a result of the experiment, a 0.4 nm spaced 130- channel demultiplexer with a channel uniformity of 3.5 dB, a 3 dB bandwidth of 0.12 nm, and channel cross talk of -20 dB is experimentally achieved. PMID- 17119568 TI - Spatial carrier fringe pattern demodulation by use of a two-dimensional continuous wavelet transform. AB - A novel technique that uses a fan two-dimensional (2D) continuous wavelet transform (CWT) to phase demodulate fringe patterns is proposed. The fan 2D CWT algorithm is tested by using computer generated and real fringe patterns. The result of this investigation reveals that the 2D CWT technique is capable of successfully demodulating fringe patterns. The proposed algorithm demodulates fringe patterns without the requirement of removing their background illumination prior to the demodulation process. Also, the algorithm is exceptionally robust against speckle noise. The performance of the 2D CWT technique in fringe pattern demodulation is compared with that of the 1D CWT algorithms. This comparison indicates that the 2D CWT outperforms its 1D counterpart for this application. PMID- 17119569 TI - Speckle activity images based on the spatial variance of the phase. AB - We propose the display of the local spatial variance of the temporal variations of the phase as an activity descriptor in dynamic speckle images. The spatial autocorrelation of the speckle intensity is calculated in sliding windows, and an estimation of the variance of the phase variations in each region of the sample is determined. The activity images obtained in this way depict some interesting features and in some cases they could be related to physical magnitudes in the samples. A simulation is presented, and examples corresponding to usual study cases are also shown, namely, fruit bruising and paint drying. PMID- 17119570 TI - Vectorial modeling of near-field imaging with uncoated fiber probes: transfer function and resolving power. AB - Using exact 3D vectorial simulations of radiation coupling into uncoated dielectric fiber probes, we calculate amplitude transfer functions for conical single-mode fiber tips at the light wavelength of 633 nm. The coupling efficiency of glass fiber tips is determined in a wide range of spatial frequencies of the incident radiation for opening angles varying from 30 degrees to 120 degrees . The resolution in near-field imaging with these tips is considered for field distributions limited in both direct and spatial-frequency space. The characteristics of the transfer functions describing the relation between probed optical fields and near-field images are analyzed in detail. The importance of utilizing a perfectly sharp tip is also examined. PMID- 17119571 TI - Encrypted holographic memory using an encoded reference wave. AB - A secure holographic memory system is proposed by use of an encoded reference beam. The reference beam is encrypted by a fiber-optic faceplate, which serves as a phase mask. There are seven keys in the system including the position and direction of the fiber bundle and the direction of the incident beam. The experiment shows that the total key length is larger than 1.8 x 1019. The method can be used directly in a shift-multiplexing system with high selectivity. PMID- 17119572 TI - Bidirectional reflectance study on dry, wet, and submerged particulate layers: effects of pore liquid refractive index and translucent particle concentrations. AB - We performed extensive bidirectional reflectance measurements on dry, wet, and submerged particulate layers with various albedos to investigate the darkening effect caused by wetting with fluids. It was found that, in addition to the reduction of the refractive index contrast when there is a pore liquid (wetted), the concentration of translucent grains in a particulate layer and the surface roughness conditions of the individual grains make important contributions to the wetting-induced darkening effect. Reflectance measurements on glass-sediment mixtures confirmed that, as the concentration of translucent particles increases, the reflectance of the dry layers increases while that of the wetted layers decreases. Measurements indicate that neither the prediction made by the theory of Twomey et al. [Appl. Opt. 25, 431 (1986)] nor that of Lekner and Dorf [Appl. Opt. 27, 1278 (1988)] is sufficient. PMID- 17119573 TI - Transmission medium and full fiber-optic setup for quantum key distribution applications. AB - A fiber-optic-based coupled waveguide transmission medium is proposed to distribute secret keys in a single-photon polarization-based quantum cryptography setup. Polarization maintenance properties and coupling phenomena of the transmission medium are exploited to achieve accuracy and security of the transferred key. Elliptic fibers and fiber couplers are used to prepare the transmitted photons at the sender as well as analyze them at the receiver. The uniqueness of the setup stands on the exclusive use of fiber-optic components, enabling its construction on a single fiber line. PMID- 17119574 TI - External-cavity wavelength tunable laser with an electro-optic deflector. AB - A near-IR tunable external-cavity laser that uses an electro-optic deflector and a reflection grating as a tunable filter is demonstrated. The deflector is an electronically reconfigurable nonpixelated nematic liquid-crystal device. By controlling the voltage applied to the electro-optic deflector, a tuning range of 12 nm and a side-mode suppression higher than 30 dB are obtained. PMID- 17119575 TI - Modified negative-branch confocal unstable resonator. AB - A new type of unstable resonator, suitable for a laser with a large medium cross section and a small or median output coupling, is presented. The resonator configuration, a modification of a negative-branch confocal unstable resonator, is numerically investigated. The basis of the theory is the Fresnel-Kirchhoff integral equation, and the calculations describe a passive resonator. With respect to output mirror tilting, the calculations confirm that the modified negative-branch confocal unstable resonator is less sensitive to mirror misalignments than the conventional negative-branch confocal unstable resonator. Furthermore, the modified resonator improves the beam quality in comparison with the conventional unstable resonator. PMID- 17119576 TI - Determination of slope in lidar data using a duplicate of the inverted function. AB - An iterative method for determining slope in noisy lidar data is considered based on the use of a corrected ('shaped') inverted function and an assumed behavior of the unknown function of interest (an 'image function'). The method is utilized for extracting extinction- coefficient profiles from data of multiangle measurements. The sequence and specifics of the retrieval procedure, results of simulations, and essentials of the practical retrieval of particulate extinction coefficient profiles from signals of the elastic scanning lidar are considered. The methodology may be applicable when extracting the extinction-coefficient profiles from an elastic lidar operating in a multiangle scanning mode, a combined Raman elastic-backscatter lidar, or a high spectral resolution lidar operating in a fixed angular position. PMID- 17119577 TI - Effect of aerosol microphysical properties on polarization of skylight: sensitivity study and measurements. AB - We analyze the sensitivity of the degree of linear polarization in the Sun's principal plane as a function of aerosol microphysical parameters: the real and imaginary parts of the refractive index, the median radius and geometric standard deviation of the bimodal size distribution (both fine and coarse modes), and the relative number weight of the fine mode at a wavelength of 675 nm. We use Mie theory for single-scattering simulations and the doubling-adding method with the inclusion of polarization for multiple scattering. It is shown that the behavior of the degree of linear polarization is highly sensitive to both the small mode of the bimodal size distribution and the real part of the refractive index of aerosols, as well as to the aerosol optical thickness; whereas not all parameters influence the polarization equally. A classification of the importance of the input parameters is given. This sensitivity study is applied to an analysis of ground-based polarization measurements. For the passive remote sensing of microphysical and optical properties of aerosols, a ground-based spectral polarization measuring system was built, which aims to measure the Stokes parameters I, Q, and U in the visible (from 410 to 789 nm) and near-infrared (from 674 to 995 nm) spectral range with a spectral resolution of 7 nm in the visible and 2.4 nm in the near infrared. We compare polarization measurements taken with radiative transfer simulations under both clear- and hazy-sky conditions in an urban area (Cabauw, The Netherlands, 51.58 degrees N, 4.56 degrees E). Conclusions about the microphysical properties of aerosol are drawn from the comparison. PMID- 17119578 TI - Laser-induced fluorescence-cued, laser-induced breakdown spectroscopy biological agent detection. AB - Methods for accurately characterizing aerosols are required for detecting biological warfare agents. Currently, fluorescence-based biological agent sensors provide adequate detection sensitivity but suffer from high false-alarm rates. Combining single-particle fluorescence analysis with laser-induced breakdown spectroscopy (LIBS) provides additional discrimination and potentially reduces false-alarm rates. A transportable UV laser-induced fluorescence-cued LIBS test bed has been developed and used to evaluate the utility of LIBS for biological agent detection. Analysis of these data indicates that LIBS adds discrimination capability to fluorescence-based biological-agent detectors. However, the data also show that LIBS signatures of biological agent simulants are affected by washing. This may limit the specificity of LIBS and narrow the scope of its applicability in biological-agent detection. PMID- 17119579 TI - Enhancing near-infrared avalanche photodiode performance by femtosecond laser microstructuring. AB - A processing technique using femtosecond laser pulses to microstructure the surface of a silicon avalanche photodiode (APD) has been used to enhance its near infrared (near-IR) response. Experiments were performed on a series of APDs and APD arrays using various structuring parameters and poststructuring annealing sequences. Following thermal annealing, we were able to fabricate APD arrays with quantum efficiencies as high as 58% at 1064 nm without degradation of their noise or gain performance. Experimental results provided evidence to suggest that the improvement in charge collection is a result of increased absorption in the near IR. PMID- 17119580 TI - Counting signal processing and counting level normalization techniques of polarization-insensitive fiber-optic Michelson interferometric sensors. AB - A counting signal processing technique of the fiber-optic interferometric sensor is proposed. The technique is capable of counting the numbers of the maximum and minimum of the output interferometric signal in a specific time duration, and it can be used as the basis to distinguish the sensing phase signal. It can also be used as a signal detector on applications such as intrusion detection. All sensors are subject to aging of the optical components and bending loss, and therefore the output signal of each sensor may vary with time. We propose a counting level normalization technique to compensate for these variations and to obtain the correct counting numbers. PMID- 17119581 TI - Grinding surface roughness measurement based on the co-occurrence matrix of speckle pattern texture. AB - Surface speckle pattern intensity distribution resulting from laser light scattering from a rough surface contains various information about the surface geometrical and physical properties. A surface roughness measurement technique based on the texture analysis of surface speckle pattern texture images is put forward. In the surface roughness measurement technique, the speckle pattern texture images are taken by a simple setup configuration consisting of a laser and a CCD camera. Our experimental results show that the surface roughness contained in the surface speckle pattern texture images has a good monotonic relationship with their energy feature of the gray-level co-occurrence matrices. After the measurement system is calibrated by a standard surface roughness specimen, the surface roughness of the object surface composed of the same material and machined by the same method as the standard specimen surface can be evaluated from a single speckle pattern texture image. The robustness of the characterization of speckle pattern texture for surface roughness is also discussed. Thus the surface roughness measurement technique can be used for an in process surface measurement. PMID- 17119582 TI - Phase unwrapping method for three-dimensional stress analysis by scattered-light photoelasticity with unpolarized light. AB - In scattered-light photoelasticity with unpolarized light, the secondary principal stress direction psi and the relative phase retardation rho in a three dimensional stressed model with rotation of the principal stress axes can be obtained by use of Stokes parameters. For completely automated stress analysis, measurements of the total relative phase retardation and the secondary principal stress direction over the entire field are required, and it is necessary to unwrap psi and rho. A phase unwrapping method is thus proposed for the determination of these values based on scattered-light photoelasticity. The values are easily obtained via an arctangent function, overcoming the error associated with the quarter-wave plate by employing an incident light of different wavelengths. The proposed technique provides automated and nondestructive determination of the total relative phase retardation and the secondary principal stress direction in a model exhibiting rotation of the principal stress axes. PMID- 17119583 TI - Accurate wavelength calibration in the near-infrared for multielement analysis without the need for reference spectra. AB - A new technique has been developed for an instant, precise, and accurate wavelength calibration over a wide pixel array for simultaneous, multielement spectral analysis based on an inverse numerical solution to the grating dispersion function. This technique is applicable to multielement analytical applications such as laser-induced breakdown spectroscopy (LIBS), particularly when using high-density gratings in the upper visible and in the near-infrared, where nonmetallic elements are detected. This application overcomes the need to use reference spectra for each window of observation and is tested on a commercially available LIBS instrument. PMID- 17119584 TI - Phase-step calibration technique based on a two-run-times-two-frame phase-shift method. AB - A novel phase-step calibration technique is presented on the basis of a two-run times-two-frame phase-shift method. First the symmetry factor M is defined to describe the distribution property of the distorted phase due to phase-shifter miscalibration; then the phase-step calibration technique, in which two sets of two interferograms with a straight fringe pattern are recorded and the phase step is obtained by calculating M of the wrapped phase map, is developed. With this technique, a good mirror is required, but no uniform illumination is needed and no complex mathematical operation is involved. This technique can be carried out in situ and is applicable to any phase shifter, whether linear or nonlinear. PMID- 17119585 TI - Photonic crystal waveguide-based Mach-Zehnder demultiplexer. AB - A Mach-Zehnder demultiplexer with two different waveguide arms is proposed and studied theoretically in photonic crystal. The two waveguide arms with different widths function as a phase shifter. The operating wavelength spacing depends on the length of the two waveguide arms. The photonic bandgap is calculated by the plane-wave expansion method, and the device is simulated by the finite-difference time-domain method. PMID- 17119586 TI - Modeling of optical properties of silver-doped nanocomposite glasses modified by electric-field-assisted dissolution of nanoparticles. AB - It has been demonstrated recently that silver nanoparticles embedded in a glass matrix can be dissolved by the combination of an intense dc electric field and moderately elevated temperature. In an intermediate state of this process percolated silver layers inside the glass can also occur. These structural modifications significantly modify the optical behavior of the glass, suggesting an interesting perspective for the engineering of optical properties of this kind of metallodielectric materials. We present the optical characterization of silver doped glasses subjected to the electric-field-assisted dissolution of nanoparticles. The characterization is performed by means of fitting spectrophotometric measurements. The optical properties of the investigated samples are described in terms of the interference between the light reflected from the glass surface and the light reflected from a buried silver-containing layer formed in the depths of the glass. The analysis of the data reveals a porosity of the glass in the region where the nanoparticles are dissolved that can be attributed to the presence of residual nanopores. PMID- 17119587 TI - High-efficiency collector design for extreme-ultraviolet and x-ray applications. AB - A design of a two-reflection mirror for nested grazing-incidence optics is proposed in which maximum overall reflectivity is achieved by making the two grazing-incidence angles equal for each ray. The design is proposed mainly for application to nonimaging collector optics for extreme-ultraviolet microlithography where the radiation emitted from a hot plasma source needs to be collected and focused on the illuminator optics. For completeness, the design of a double- reflection mirror with equal reflection angles is also briefly outlined for the case of an object at infinity for possible use in x-ray applications. PMID- 17119588 TI - Resolution in optically addressed spatial light modulators based on dye-doped liquid crystals. AB - Dye-doped nematic liquid crystals (LCs) are studied as materials for single-layer optically addressed spatial light modulators. The dopant is 2,5-azo-substituted anthraquinone (ASAQ) dye. The resolution in the ASAQ-doped LC systems does not depend on the device thickness (in the 5-125 microm range). The efficiency increases with the increase of the thickness and begins to saturate in devices thicker than 40 microm. The limiting resolution in the thick devices is 400 line pairs per millimeter. The limitations of performance (efficiency and resolution) in the studied systems are discussed. PMID- 17119589 TI - Comparison of annular wavefront interpretation with Zernike circle polynomials and annular polynomials. AB - A general wavefront fitting procedure with Zernike annular polynomials for circular and annular pupils is proposed. For interferometric data of typical annular wavefronts with smaller and larger obscuration ratios, the results fitted with Zernike annular polynomials are compared with those of Zernike circle polynomials. Data are provided demonstrating that the annular wavefront expressed with Zernike annular polynomials is more accurate and meaningful for the decomposition of aberrations, the calculation of Seidel aberrations, and the removal of misalignments in interferometry. The primary limitations of current interferogram reduction software with Zernike circle polynomials in analyzing wavefronts of annular pupils are further illustrated, and some reasonable explanations are provided. It is suggested that the use of orthogonal basis functions on the pupils of the wavefronts analyzed is more appropriate. PMID- 17119590 TI - Global beam shaping with nonuniformly polarized beams: a proposal. AB - A procedure for global beam shaping by modifying some global spatial parameters characteristic of the beam is proposed. This method is based on the generation of a nonuniformly polarized beam using a Mach-Zehnder system with two suitably shaped intensity transmittances and orthogonal linear polarizers. The changes in beam quality and kurtosis parameters after a linear polarizer placed at the output of the system are investigated. PMID- 17119591 TI - Achromatic half-wave plate for submillimeter instruments in cosmic microwave background astronomy: modeling and simulation. AB - We adopted an existing formalism and modified it to simulate, with high precision, the transmission, reflection, and absorption of multiple-plate birefringent devices as a function of frequency. To validate the model, we use it to compare the measured properties of an achromatic five-plate device with a broadband antireflection coating to expectations derived from the material optical constants and its geometric configuration. The half-wave plate presented here is observed to perform well with a phase shift variation of < 2 degrees from the ideal 180 degrees over a bandwidth of Deltav/v approximately 1 at millimeter wavelengths. This formalism represents a powerful design tool for birefringent polarization modulators and enables its optical properties to be specified with high accuracy. PMID- 17119592 TI - Thin-film coatings--a transmission ellipsometric function approach: I. Nonnegative transmission systems, polarization devices, coatings, and closed-form design formulas. AB - The transmission ellipsometric function (TEF) of a film-substrate system relates the polarization change, upon transmission, of an electromagnetic wave obliquely incident on, and transmitted through, a film-substrate system. The behavior of the TEF depends on the category of the film-substrate system: negative, zero, or positive. The category is determined by the sign of [equation in text]: negative for a negative film-substrate system, zero for a zero system, and positive for a positive system. We discuss the behavior of the TEFs of the two transparent nonnegative film-substrate systems, zero and positive. We describe the TEF as two successive transformations and analyze its behavior as the angle of incidence and film thickness of the film-substrate system are changed. We use the constant angle-of-incidence contours and constant- thickness contours to analyze and utilize that behavior. From the analysis and understanding of the behavior of the TEF, and from the definition of a polarization device as a film-substrate system that introduces prescribed polarization changes, we discuss the design of all possible types of polarization devices using either of the two systems. We present a design formula for each. We also present a general formula that is used for the design of any of the devices. Thin-film coatings are treated as polarization devices for the purposes of our discussion. We conclude with a brief discussion of suggested practical modifications to, and simplifications of, ellipsometric memory, which is an interesting application of polarization devices for which there is a patent pending. PMID- 17119593 TI - Micropore x-ray optics using anisotropic wet etching of (110) silicon wafers. AB - To develop x-ray mirrors for micropore optics, smooth silicon (111) sidewalls obtained after anisotropic wet etching of a silicon (110) wafer were studied. A sample device with 19 microm wide (111) sidewalls was fabricated using a 220 microm thick silicon (110) wafer and potassium hydroxide solution. For what we believe to be the first time, x-ray reflection on the (111) sidewalls was detected in the angular response measurement. Compared to ray-tracing simulations, the surface roughness of the sidewalls was estimated to be 3-5 nm, which is consistent with the atomic force microscope and the surface profiler measurements. PMID- 17119594 TI - Scattering of light by a periodic structure in the presence of randomness. V. Detection of successive peaks in a periodic structure. AB - We address the problem of detecting periodic structures hidden behind roughness. We have shown that if r(0) is the coherence length of the scattered radiation, due to the random part of the surface, and Lambda is the wavelength of the periodic part of the surface, then with a matched filtering method that we introduce, and by using simple computations with the intensity data, it is possible to detect the hidden first-order peak even when (r(0)/Lambda) approximately 0.11. Here we advance the method to bring out very weak second order peaks, which we demonstrate for what we believe is the first time. The unmistakable presence of both the first- and second-order peaks, which have identical shapes as the zeroth-order peak, is strong evidence of the hidden periodicity and serves as a novel method for the detection of weak periodicities hidden behind strong randomness. PMID- 17119595 TI - Fast acquisition system for digital holograms and image processing for three dimensional display with data manipulation. AB - A three-dimensional (3D) digital holographic display system with image processing is presented. By use of phase-shifting digital holography, we obtain the complex amplitude of a 3D object at a recording plane. Image processing techniques are introduced to improve the quality of the reconstructed 3D object or manipulate 3D objects for elimination and addition of information by modifying the complex amplitude. The results show that the information processing is effective in such manipulations of 3D objects. We also show a fast recording system of 3D objects based on phase-shifting digital holography for display with image processing. The acquisition of 3D object information at 500 Hz is demonstrated experimentally. PMID- 17119596 TI - Aerosol polarization effects on atmospheric correction and aerosol retrievals in ocean color remote sensing. AB - The current ocean color data processing system for the Sea-viewing Wide Field-of View Sensor (SeaWiFS) and the moderate resolution imaging spectroradiometer (MODIS) uses the Rayleigh lookup tables that were generated using the vector radiative transfer theory with inclusion of the polarization effects. The polarization effects, however, are not accounted for in the aerosol lookup tables for the ocean color data processing. I describe a study of the aerosol polarization effects on the atmospheric correction and aerosol retrieval algorithms in the ocean color remote sensing. Using an efficient method for the multiple vector radiative transfer computations, aerosol lookup tables that include polarization effects are generated. Simulations have been carried out to evaluate the aerosol polarization effects on the derived ocean color and aerosol products for all possible solar-sensor geometries and the various aerosol optical properties. Furthermore, the new aerosol lookup tables have been implemented in the SeaWiFS data processing system and extensively tested and evaluated with SeaWiFS regional and global measurements. Results show that in open oceans (maritime environment), the aerosol polarization effects on the ocean color and aerosol products are usually negligible, while there are some noticeable effects on the derived products in the coastal regions with nonmaritime aerosols. PMID- 17119597 TI - Adaptive optics sky coverage modeling for extremely large telescopes. AB - A Monte Carlo sky coverage model for laser guide star adaptive optics systems was proposed by Clare and Ellerbroek [J. Opt. Soc. Am. A 23, 418 (2006)]. We refine the model to include (i) natural guide star (NGS) statistics using published star count models, (ii) noise on the NGS measurements, (iii) the effect of telescope wind shake, (iv) a model for how the Strehl and hence NGS wavefront sensor measurement noise varies across the field, (v) the focus error due to imperfectly tracking the range to the sodium layer, (vi) the mechanical bandwidths of the tip tilt (TT) stage and deformable mirror actuators, and (vii) temporal filtering of the NGS measurements to balance errors due to noise and servo lag. From this model, we are able to generate a TT error budget for the Thirty Meter Telescope facility narrow-field infrared adaptive optics system (NFIRAOS) and perform several design trade studies. With the current NFIRAOS design, the median TT error at the galactic pole with median seeing is calculated to be 65 nm or 1.8 mas rms. PMID- 17119598 TI - Isotope-selective excitation of 41Ca isotope in Doppler-free two-photon continuous-wave excitation: a case study. AB - Seven schemes are studied theoretically for Doppler-free two-photon excitation of rare (41)Ca isotope using single-mode continuous-wave lasers. The ionization efficiencies and optical selectivities for all the schemes are calculated for various powers of the excitation and ionization lasers and for various focusing conditions of the two lasers. To maximize the ionization efficiencies and the optical selectivities, wavelength-dependent Stark compensation is used. Certain laser wavelengths of the ionization step termed as magic wavelengths are identified for compensating the Stark shift induced by the excitation laser. The effects of the Stark-shift-induced asymmetry and its reversal by selecting the appropriate magic wavelength for the ionization step for various excitation and ionization laser intensities are investigated. The ionization efficiency and optical selectivity for the best scheme after Stark compensation are found to be 8.4 x 10(-4) and approximately 9 x 10(3), respectively. PMID- 17119599 TI - Design and analysis of a flow-through integrating cavity absorption meter. AB - We present a design for a flow-through integrating cavity absorption meter. This instrument, in principle, is capable of measuring the spectral optical absorption coefficient of natural waters in situ independently of scattering effects. Monte Carlo simulations are used to determine the design parameters and evaluate instrument performance. We investigate both detector response and the distribution of radiant energy inside the instrument and present empirical equations describing these quantities as a function of the absorption coefficient. The effects of changing the instrument geometry are illustrated. Finally, we discuss the effects of scattering on the instrument performance and verify that they are negligible for natural waters. PMID- 17119600 TI - Transverse-mode control in vertical-cavity surface-emitting lasers via a patterned phase aperture. AB - Transverse-mode discrimination in vertical-cavity surface-emitting lasers that contain a patterned phase aperture is analyzed numerically. The two lowest-order modes are calculated for different aperture shapes and sizes. They are then expanded in Laguerre-Gaussian modes to study the power distribution as well as their beam propagation factor. The mode selection depends on the aperture's size and degree of symmetry. The maximum value for the mode discrimination in the case of a specific phase aperture is determined, and an enhancement by a factor of 3, compared to the case without a phase aperture, is found. PMID- 17119601 TI - Gain measurements of Fabry-Perot InP/InGaAsP lasers using an ultrahigh-resolution spectrometer. AB - Measurements of the optical gain in a semiconductor laser using a 20 MHz resolution optical spectrum analyzer are presented for what is believed to be the first time. The high resolution allows for accurate gain measurements close to the lasing threshold. This is demonstrated by gain measurements on a bulk InGaAsP 1.5 microm Fabry-Perot laser. Combined with direct measurement of transparency carrier density values, parameters were determined for characterizing the gain at a range of wavelengths and temperatures. The necessity of the use of a logarithmic gain model is shown. PMID- 17119602 TI - Extending the continuous tuning range of an external-cavity diode laser. AB - The continuous tuning range of an external-cavity diode laser can be extended by making small corrections to the external-cavity length through an electronic feedback loop so that the cavity resonance condition is maintained as the laser wavelength is tuned. By maintaining the cavity resonance condition as the laser is tuned, the mode hops that typically limit the continuous tuning range of the external-cavity diode laser are eliminated. We present the design of a simple external-cavity diode laser based on the Littman-Metcalf external-cavity configuration that has a measured continuous tuning range of 1 GHz without an electronic feedback loop. To include the electronic feedback loop, a small sinusoidal signal is added to the drive current of the laser diode creating a small oscillation of the laser power. By comparing the phase of the modulated optical power with the phase of the sinusoidal drive signal using a lock-in amplifier, an error signal is created and used in an electronic feedback loop to control the external-cavity length. With electronic feedback, we find that the continuous tuning range can be extended to over 65 GHz. This occurs because the electronic feedback maintains the cavity resonance condition as the laser is tuned. An experimental demonstration of this extended tuning range is presented in which the external-cavity diode laser is tuned through an absorption feature of diatomic oxygen near 760 nm. PMID- 17119603 TI - Feasibility study for joint retrieval of air density and ozone in the stratosphere and mesosphere with the limb-scan technique. AB - Air density is a key sensing object for its high variability especially in the mesosphere, and ozone plays an important role in the physical, chemical, and radiant processes in the atmosphere system. Therefore it is essential to obtain their global vertical distributions jointly with high precision and vertical resolution. There is little work on joint retrieval of air density and ozone distributions using the ultraviolet limb-scan technique, although much work has been done on ozone measurement. Numerical simulations of joint air density and ozone retrieval in the middle atmosphere (20-90 km) are carried out using limb scattered radiances at four bands (255, 300, 320, and 340 nm). Results show that joint retrieval of dual parameters using the limb-scan technique is feasible with high precision in nearly the whole region concerned, where air density and ozone have a precision of 1%-2% and 3%-5%, respectively, provided that high measurement precision and accurate correction of multiple-scattered radiance at long ultraviolet bands are ensured. PMID- 17119605 TI - Population structure of the Brazilian southern green stink bug, Nezara viridula. AB - The Southern Green Stink Bug, Nezara viridula (L.) (Heteroptera: Pentatomidae), is a cosmopolitan and economically important pest to several crops. Studies on N. viridula migration and population structure have been neglected. We studied geographically distinct Brazilian N. viridula populations to assess their variability and to determine gene flow among them. DNA from specimens collected on soybean fields were subjected to RAPD analysis to determine genetic similarity and population structure parameters. All N. viridula populations studied were genetically distinct from the others. The maximum similarity occurred between populations from Londrina and Sertanopolis (Parana State). The Cruz Alta population was the most divergent from the others. Despite the short distance between Cambe and Londrina (ca. 29 km), and the absence of geographic barriers, both populations clustered in different groups and the estimated gene flow index (Nm) among them was 2.02, indicating relatively restricted migration. The estimated overall index, Nm was 1.41 suggesting that N. viridula is a better flier than the Neotropical Brown stink bug, Euschistus heros (Nm =0.83). PMID- 17119606 TI - A synthesis of the temperature dependent development rate for the obliquebanded leafroller, Choristoneura rosaceana. AB - The obliquebanded leafroller, Choristoneura rosaceana (Harris) (Lepidoptera: Tortricidae), is a serious pest of apples and other tree crops throughout North America. A review of temperature dependent development and models show that five different lower thresholds for development are published and used as the basis of heat-driven phenology models. We present a small lab data set of C. rosaceana development at four different temperatures and combine this with literature-based data into a single meta-analysis. Our analysis shows that the data from the different studies can be lumped together and the combined analysis suggests the lower and upper thresholds for development from egg to adult are approximately 10 degrees C and 30 degrees C, respectively. PMID- 17119607 TI - The biology and behavior of the longhorned beetle, Dectes texanus on sunflower and soybean. AB - The biology and behavior of the longhorned beetle Dectes texanus LeConte (Coleoptera: Cerambycidae) was studied on two host plants that suffer economic losses from this pest; sunflower, Helianthus annuus, and soybean, Glycines max. Reciprocal crosses of D. texanus collected from the two plants all produced viable progeny, indicating that conspecific insects attack both crops. Pupae from soybean stalks weighed about 40% less than those from sunflower, and adults fed on soybean lived a mean of 23 days, compared to a mean of 53 days (males) and 76 days (females) for those fed sunflower. A female's larval host plant had no effect on her tendency to ovipuncture plants of either type in a greenhouse trial. A field-tested population collected exclusively from sunflower contained three types of females in similar proportions: those that laid eggs only on sunflower, those that laid only on soybean, and those that laid equally on both host plants. Females in field trials fed more on the plant they had fed on in the laboratory, but soybean-fed females fed more on soybean than did sunflower-fed females. Females fed soybean also made more ovipunctures on soybean plants in field trials than sunflower-fed females, but their responses to sunflower plants were similar. Females displayed higher total ovipositional activity when they encountered sunflower first in the field, and lower total activity when they encountered soybean first. Feeding scores were significantly correlated with ovipunctures and eggs on both plant types. We conclude that sunflower is the preferred host plant, although females will accept soybean when it is the only available food. The results suggest that D. texanus is still in the initial stages of a host range expansion with female host selection behavior demonstrating both genetic influences and phenotypic flexibility. Sunflower represents a nutritionally superior, ancestral host plant and relatively high fitness costs are still associated with utilization of the novel host plant, soybean, costs that may be offset by benefits such as reduced intraspecific competition. These potential benefits and their consequent implications for D. texanus host range evolution are hypothesized and discussed. PMID- 17119609 TI - Evidence of sexual reproduction of woolly apple aphid, Eriosoma lanigerum, in New Zealand. AB - Reproduction of the woolly apple aphid, Eriosoma lanigerum (Hausmann), can take place parthenogenetically or sexually when both host plants, apple (Malus domestica Borkh.) and elm (Ulmus americana L.) are available. Since elm is not commonly grown in New Zealand, E. lanigerum, a major pest of apple, is thought to reproduce only parthenogenetically here. During our studies between 1999 and 2003, different morphs of E. lanigerum have been observed on apple trees, which were studied in more detail in 2003 and 2004. In the laboratory, alates reproduced mainly sexual morphs with degenerated mouthparts. Oviparous females lived for 8.95 +/- 0.17 days (n = 20) and males lived for 7.1 +/- 0.12 days (n = 20). Both went through 4 moults, without feeding or changing body size. Oviparous female laid a single egg and died soon after oviposition. In addition to sexual morphs, shiny brown, oblong eggs were seen on apple leaves grown outside as well as in the glasshouse. As the numbers of eggs and sexual morphs on trees grown outside were less than on those grown in the glasshouse, we suggest that alates disperse into the natural environment searching for an apple or elm tree to continue the sexual reproduction while spreading the population. PMID- 17119608 TI - Identification and partial characterization of the enzyme of omega: one of five putative DPP IV genes in Drosophila melanogaster. AB - The omega (ome) gene product is a modifier of larval cuticle protein 5 and its alleles (and duplicates) in the third instar of Drosophila melanogaster. Using deletion mapping the locus mapped to 70F-71A on the left arm of chromosome 3. A homozygote null mutant (ome 1) shows a pleiotropic phenotype that affected the size, developmental time of the flies, and the fertility (or perhaps the behavior) of homozygous mutant males. The omega gene was verified as producing a dipeptidyl peptidase IV (DPPIV) by genetic analysis, substrate specificity and pH optimum. The identity of the gene was confirmed as CG32145 (cytology 70F4) in the Celera Database (Berkeley Drosophila Genome Project), which is consistent with its deletion map position. The genomic structure of the gene is described and the decrease in DPPIV activity in the mutant ome1 is shown to be due to the gene CG32145 (omega). The D. melanogaster omega DPPIV enzyme was partially purified and characterized. The exons of the ome1 mutant were sequenced and a base substitution mutation in exon 4 was identified that would yield a truncated protein caused by a stop codon. A preliminary study of the compartmentalization of the omega DPPIV enzyme in several organs is also reported. PMID- 17119610 TI - Preliminary survey of the mayflies (Ephemeroptera) and caddisflies (Trichoptera) of Big Bend Ranch State Park and Big Bend National Park. AB - The mayfly (Insecta: Ephemeroptera) and caddisfly (Insecta: Trichoptera) fauna of Big Bend National Park and Big Bend Ranch State Park are reported based upon numerous records. For mayflies, sixteen species representing four families and twelve genera are reported. By comparison, thirty-five species of caddisflies were collected during this study representing seventeen genera and nine families. Although the Rio Grande supports the greatest diversity of mayflies (n=9) and caddisflies (n=14), numerous spring-fed creeks throughout the park also support a wide variety of species. A general lack of data on the distribution and abundance of invertebrates in Big Bend National and State Park is discussed, along with the importance of continuing this type of research. PMID- 17119611 TI - The shield-backed bug, Pachycoris stallii: description of immature stages, effect of maternal care on nymphs, and notes on life history. AB - The life history of the shield-backed bug, Pachycoris stallii Uhler (Heteroptera: Scutelleridae), immatures was studied on its host plant, Croton californicus Muell.-Arg. (Euphorbiaceae), in Baja California Sur, Mexico. Immature stages are described and illustrated. Pachycoris stallii is bi- or multivoltine and occurs in xeric areas with sandy soil where it is rarely encountered away from C. californicus. Nymphs and adults feed on seeds within C. californicus fruit. Bugs oviposit on the underside of leaves, and females guard their eggs and first instar nymphs from natural enemies. Embryonic orientation of prolarvae is nonrandom; each embryo is oriented with its venter directed toward the ground. This orientation may facilitate aggregation of first instars. The longitudinal axes of eggs are always oriented upward at about a 16 degree angle of deviation from a line perpendicular to the leaf surface. This is the first recorded observation of this phenomenon in Pentatomoidea. Experimental removal of females guarding first instars results in 100% loss of nymphs, and this is attributed to disruption of the aggregative behavior of nymphs. Maternal guarding appears to be a net benefit to P. stallii, despite possible costs to the brooding female. PMID- 17119612 TI - Effects of Wolbachia in the uzifly, Exorista sorbillans, a parasitoid of the silkworm, Bombyx mori. AB - The uzifly, Exorista sorbillans (Diptera: Tachinidae), a parasitoid of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), harbours Wolbachia (Rickettsia) endosymbionts. Administration of 0.05 mg/ml oxytetracycline to the adult uziflies removed Wolbachia endosymbionts and resulted in different reproductive disorders, such as i) reduction in fecundity of uninfected females, ii) cytoplasmic incompatibility in crosses between infected males and uninfected females, iii) sterility in the crosses between both males and females from uninfected populations, and iv) sex-ratio distortion in uninfected females irrespective of the presence of Wolbachia in males. However, tetracycline treatment did not have much effect on longevity of the uzifly. These results suggest that the interaction of Wolbachia with its uzifly host is one of mutual symbiosis as it controls the reproductive physiology of its hosts. PMID- 17119613 TI - Sex differences in frass production and weight change in Tenebrio molitor (Coleoptera) infected with cysticercoids of the tapeworm Hymenolepis diminuta (Cestoda). AB - In their intermediate host, parasites alter aspects of host physiology including waste production and body weight. Further, this alteration may differ between female and male hosts. To study this, a beetle (Tenebrio molitor)-tapeworm (Hymenolepis diminuta) system was used. Infected and uninfected male and female beetles were individually housed in vials without food. Each beetle's weight change and frass production were measured over 24 h periods at 3, 7, 12 and 16 days post-infection. Treatment (infection) had no effect on weight change, but males lost more weight than females. Further, infected females produced more frass than control females. Males on the day of infection had a higher food intake than females. These results suggest that males will be more exposed to infection than females and could explain why males had a higher median cysticercoid infection level. PMID- 17119614 TI - The broadly insecticidal Photorhabdus luminescens toxin complex a (Tca): activity against the Colorado potato beetle, Leptinotarsa decemlineata, and sweet potato whitefly, Bemisia tabaci. AB - Toxin complex a (Tca), a high molecular weight insecticidal protein complex produced by the entomopathogenic bacterium Photorhabdus luminescens, has been found to be orally toxic to both the Colorado potato beetle, Leptinotarsa decemlineata, and the sweet potato whitefly, Bemisia tabaci biotype B. The 48 hour LC50 for Tca against neonate L. decemlineata was found to be 2.7 ppm, and the growth of 2nd instar L. decemlineata exposed to Tca for 72 hours was almost entirely inhibited at concentrations above 0.5 ppm. B. tabaci was highly susceptible to Tca as well; newly emerged nymphs that were artificially fed Tca developed poorly, or not at all. Tca concentrations between 0.1 and 0.2 ppm reduced the number of nymphs reaching the second instar by 50%. In addition, a preparation of Tca missing two prominent subunits, TcaAii and TcaAiii, was found to be at least as toxic to L. decemlineata and B. tabaci as Tca itself, indicating that the activity of Tca is not dependant on the presence of these subunits at the time of ingestion. PMID- 17119615 TI - Reproductive strategies of Tribolium flour beetles. AB - Although, beetles of the genus Tribolium first evolved as saprophylic insects, they have adapted to the stored products environment for several thousand years. In this study reproductive strategies are described for eight species of Tribolium that are known to occur in this environment. Experiments were conducted under the same conditions for every species, and several life history traits, including egg mass, adult mass, developmental time and fecundity were examined and compared among these species. Common reproductive strategies were not found among the different species and univariate analysis highlighted strong differences between the species for most of the traits investigated. Some species showed reproductive traits that are likely to give a fitness advantage in the environment of stored products. Multivariate statistical analysis allowed the detection of different sub-groups with respect to their reproductive strategy. Adult mass and egg-to-adult developmental time discriminated between groups. Intraspecific allometric relationships were further investigated but only a few correlations appeared to be significant. PMID- 17119616 TI - The effect of water on the ground nesting habits of the giant tropical ant, Paraponera clavata. AB - The large predatory ant, Paraponera clavata, exerts measurable top-down effects in wet and moist Neotropical forests, and therefore its distribution has potential ecological implications. To determine how water affects the presence of this important predator, the ground nesting ecology of P. clavata was examined with respect to various habitat characteristics. Four hectares of disturbed Costa Rican lowland rain forest were surveyed for ant colonies to determine nest distribution patterns in wet and dry habitat; significantly more colonies were found in dry habitat. Seventeen of 19 nests built on slopes of > 5 degrees inclination were positioned on the downward side of the tree, possibly using the trunk as a shield against runoff during rain showers. Moisture and pH inside nests were significantly different from adjacent soil. These results suggest that water influences the ground nesting habits of P. clavata, thus ecological differences between comparatively wet and dry portions of tropical forests may arise from the relative abundance of this ant species. PMID- 17119617 TI - Revision of Heterocloeon McDunnough (Ephemeroptera: Baetidae). AB - The North American genus Heterocloeon McDunnough is redefined and shown to be distinct from other two-tailed Baetis complex genera, including Acentrella Bengtsson and Plauditus Lugo-Ortiz and McCafferty, which are also represented in North America. Heterocloeon is divided into three distinct subgenera, including Heterocloeon s.s., Iswaeon McCafferty and Webb, new subgenus, and Jubilatum McCafferty and Jacobus, new subgenus. Cladistic analysis supports the recognition of these subgenera within a genus defined by the presence of an apomorphic secondary ridge or denticle row on the larval claw; Heterocloeon s.s. and Jubilatum being sister groups, with their stem and Iswaeon branching basally within the Heterocloeon clade. The subgenus Heterocloeon, which possesses larval prothoracic osmobranchiae, retains the traditional species makeup of the genus. Iswaeon, which possesses a basally narrowed labial palp segment 2 and tibiae, and primary claw denticles of subequal length, includes a revalidated H. anoka (Daggy); H. davidi Waltz and McCafferty, new species; and H. rubrolaterale (McDunnough), new combination. Jubilatum, which posssesses an adenticulate secondary ridge on the claws and specialized tibial setation, includes H. amplum (Traver), new combination, and H. grande (Wiersema and Long), new combination. Previous confusion of H. anoka and Plaudituspunctiventris (McDunnough) is also resolved, with adult color patterns in Iswaeon and Plauditus shown to be of diagnostic importance. PMID- 17119618 TI - Effects of carbon dioxide narcosis on ovary activation and gene expression in worker honeybees, Apis mellifera. AB - In an effort to uncover genes associated with ovary activation in honey bee workers, the extent to which eight candidate genes co-varied in their expression with experimentally-induced changes in worker reproductive state was examined. Groups of caged, queenless workers narcotized with CO(2) on consecutive days early in adult life showed a significantly lower level of ovary activation than did groups of untreated workers. This same experimental treatment, by contrast, is known to accelerate ovary activation and induce egg laying in virgin honey bee queens--an observation that suggests that CO(2) narcosis has contrasting effects in queen versus worker ovary activation. Experimentally-induced changes to worker reproductive state were associated with changes in gene expression. Vitellogenin, an egg yolk precursor, and transferrin, an iron transporter, were two transcripts found to be significantly down-regulated as a function of the ovary-inhibiting treatment. CO(2) narcosis did not effect the expression of six other genes selected as putative markers for processes that may underlie ovary activation. The show that the expression of vitellogenin and transferrin is correlated with ovary activation in workers, and may therefore be part of the gene network involved in the regulatory control of functional sterility in honeybees. PMID- 17119619 TI - A new method to evaluate the biocontrol potential of single spore isolates of fungal entomopathogens. AB - Fifty Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) strains isolated from the coffee berry borer were used to develop a novel screening method aimed at selecting strains with the highest biocontrol potential. The screening method is based on percent insect mortality, average survival time, mortality distribution, percent spore germination, fungal life cycle duration, and spore production on the insect. Based on these parameters, only 11 strains merited further study. The use of a sound scientific protocol for the selection of promising fungal entomopathogens should lead to more efficient use of time, labor, and financial resources in biological control programs. PMID- 17119620 TI - AFLP markers for the R-gene in the flea beetle, Phyllotreta nemorum, conferring resistance to defenses in Barbarea vulgaris. AB - A so-called R-gene renders the yellow-striped flea beetle Phyllotreta nemorum L. (Coleoptera: Chrysomelidae: Alticinae) resistant to the defenses of the yellow rocket Barbarea vulgaris R.Br. (Brassicacea) and enables it to use it as a host plant in Denmark. In this study, genetic markers for an autosomal R-gene, inherited as a single, dominant locus in flea beetles from the Danish locality "Kvaerkeby" are described, and a genetic linkage map around this particular R gene is constructed, using the technique of AFLP (Amplified Fragment Length Polymorphism). PMID- 17119621 TI - Egg size-number trade-off and a decline in oviposition site choice quality: female Pararge aegeria butterflies pay a cost of having males present at oviposition. AB - Once mated, the optimal strategy for females of the monandrous butterfly, Pararge aegeria, is to avoid male contact and devote as much time as possible to ovipositing, as there is little advantage for females to engage in multiple matings. In other butterfly species the presence of males during egg laying has been shown to affect aspects of oviposition behavior and it has been suggested that repeated interference from males has the potential to reduce reproductive output. The aim of this study was to assess the effects of male presence during oviposition on reproductive output and behavior of a population of P. aegeria obtained from Madeira Island, Portugal, and maintained in the laboratory. Two experiments were performed where females were housed individually in small cages. Experiment 1 examined how social factors influenced the egg laying behavior of females. To do this the presence or absence of males was manipulated and egg size and number was measured over the first 14 days of oviposition. It was observed that when males were present during oviposition females made a trade-off between egg size and number. Experiment 2 examined how social factors affected oviposition site choice. Again, male presence/absence was manipulated, but in this experiment where the female laid her egg in relation to host quality was scored, and the size of the egg laid was measured. In the absence of males females selectively positioned their larger eggs on good quality host plants. However, selective oviposition was no longer observed when females were in the presence of males. We suggest that P. aegeria females from the Madeira Island population are adapted for a flexible oviposition strategy, governed by external cues, allowing a trade-off between egg size and number when the time available for egg laying is limiting. PMID- 17119622 TI - The melon fruit fly, Bactrocera cucurbitae: a review of its biology and management. AB - The melon fruit fly, Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae) is distributed widely in temperate, tropical, and sub-tropical regions of the world. It has been reported to damage 81 host plants and is a major pest of cucurbitaceous vegetables, particularly the bitter gourd (Momordica charantia), muskmelon (Cucumis melo), snap melon (C. melo var. momordica), and snake gourd (Trichosanthes anguina). The extent of losses vary between 30 to 100%, depending on the cucurbit species and the season. Its abundance increases when the temperatures fall below 32 degrees C, and the relative humidity ranges between 60 to 70%. It prefers to infest young, green, soft-skinned fruits. It inserts the eggs 2 to 4 mm deep in the fruit tissues, and the maggots feed inside the fruit. Pupation occurs in the soil at 0.5 to 15 cm below the soil surface. Keeping in view the importance of the pest and crop, melon fruit fly management could be done using local area management and wide area management. The melon fruit fly can successfully be managed over a local area by bagging fruits, field sanitation, protein baits, cue-lure traps, growing fruit fly-resistant genotypes, augmentation of biocontrol agents, and soft insecticides. The wide area management program involves the coordination of different characteristics of an insect eradication program (including local area options) over an entire area within a defensible perimeter, and subsequently protected against reinvasion by quarantine controls. Although, the sterile insect technique has been successfully used in wide area approaches, this approach needs to use more sophisticated and powerful technologies in eradication programs such as insect transgenesis and geographical information systems, which could be deployed over a wide area. Various other options for the management of fruit fly are also discussed in relation to their bio-efficacy and economics for effective management of this pest. PMID- 17119623 TI - Mitochondrial DNA variation among populations of the glassy-winged sharpshooter, Homalodisca coagulata. AB - The glassy-winged sharpshooter, Homalodisca coagulata (Homoptera: Cicadellidae), is a highly polyphagous insect species that is distributed throughout most of the southern regions of the United States. In the last 10 years, H. coagualta has become established in California and represents a significant threat to the state's 35 billion dollar wine and table grape industries. DNA sequencing analysis was used to characterize a portion of the mitochondrial cytochrome oxidase I gene from a single population of the smoke tree sharpshooter, Homalodisca liturata, in California and from 20 natural populations of H. coagulata distributed in Tahiti, California, Texas, Louisiana, Mississippi, Alabama, and Florida. The results indicate that H. liturata and H. coagulata are genetically distinct, suggesting that they do not hybridize. Populations of H. coagulata are geographically structured into two groups of haplotypes; a group of populations from east of the Mississippi River including Louisiana, Mississippi, Alabama and Florida and a group comprised of populations west of the Mississippi River from Texas and California, and from Tahiti. There was no genetic structure among haplotypes within the eastern and western groups, respectively. The data also indicates that H. coagulata in California most likely originated from a source in Texas and not from any of the populations east of the Mississippi River. PMID- 17119624 TI - Evidence for community structure and habitat partitioning in coastal dune stiletto flies at the Guadalupe-Nipomo dunes system, California. AB - This study provides empirical evidence for habitat selection by North American species of stiletto flies (Diptera: Therevidae), based on local distributions of adults and immatures, and the first hypothesis of community assemblages proposed for a stiletto fly community. Sites at three localities within the Guadalupe Nipomo dune system were sampled for stiletto flies in 1997 and 2001 by sifting sand, malaise trapping, and hand netting. Nine species were collected from four ecological zones and three intermediate ecological zones: Acrosathe novella (Coquillett), Brachylinga baccata (Loew), Nebritus powelli (Webb and Irwin), Ozodiceromyia sp., Pherocera sp., Tabudamima melanophleba (Loew), Thereva comata Loew, Thereva elizabethae Holston and Irwin, and Thereva fucata Loew. Species associations of adults and larvae with habitats and ecological zones were consistent among sites, suggesting that local distributions of coastal dune stiletto fly species are influenced by differences in habitat selection. In habitats dominated by the arroyo willow,Salix lasiolepsis, stiletto fly larvae of three species were collected in local sympatry, demonstrating that S. lasiolepsis stands along stabilized dune ridges can provide an intermediate ecological zone linking active dune and riparian habitat in the Guadalupe-Nipomo dune system. Sites dominated by European beach grass, Ammophilia arenaria, blue gum, Eucalyptus globulus, and Monterey cypress, Cupressus macrocarpa, are considered unsuitable for stiletto flies, which emphasizes the importance of terrestrial habitats with native vegetation for stiletto fly species. The local distributions of stiletto fly species at the Guadalupe-Nipomo dune system allow the community to be divided into three assemblages; active dune, pioneer scrub, and scrub riparian. These assemblages may be applicable to other coastal dune stiletto fly communities, and may have particular relevance to stiletto fly species collected in European coastal dunes. The results from this study provide a descriptive framework for studies testing habitat selection in coastal dune stiletto fly species and inform conservation of threatened dune insects. PMID- 17119625 TI - Chronobiological analysis and mass spectrometric characterization of pigment dispersing factor in the cockroach Leucophaea maderae. AB - Pigment-dispersing factor (PDF) is a neuropeptide that plays a prominent role in the circadian clock of several insects. The cockroach Leucophaea maderae was the first animal where the site of a biological clock could be located, and still is a focal point of circadian research. Although detailed studies on the action of pigment-dispersing factor and the distribution of PDF-like immunoreactivity in the L. maderae brain exist, a native pigment-dispersing factor of this species has not been characterized so far. The authentic Lem-PDF was isolated from L. maderae by a combination of high performance liquid chromatography, crab pigment dispersion bioassay and an immunosorbent assay. Mass spectrometric characterization and the conserved sequence of pigment-dispersing factor in orthopteromorphan insects suggest that Lem-PDF has the sequence NSEXINSLLGLPKVLNDAa (where X= I or L). Lem-PDF is thus identical to either Periplaneta americana PDF or Acheta domesticus PDF. Detailed analysis of PDF-like immunofluorescence in different regions of the brain suggests that there are no drastic daily changes in the amount of pigment-dispersing factor as occur in Drosophila melanogaster, which might be explained by a lack of circadian pigment dispersing factor release and production, or by phase differences between the pigment-dispersing factor neurons. PMID- 17119626 TI - Parasitism of Lygus spp. nymphs by the parasitoid wasp, Peristenus howardi, in the alfalfa seed-growing region of the Pacific Northwest. AB - The parasitoid, Peristenus howardi Shaw (Hymenoptera: Braconidae) has been found to parasitize a large proportion of Lygus species in some Washington and Idaho alfalfa seed fields. During 2002-2003 a survey was conducted to estimate the proportion of Lygus spp. (Hemiptera: Miridae) parasitized and the amountof that parasitism attributable to P. howardi in crop and non-crop plants attacked by Lygus in the alfalfa seed growing region of southwestern Idaho and eastern Oregon. Percentage parasitism was estimated from dissection of field-collected Lygus nymphs. Polymerase chain reaction (PCR) was used to amplify DNA extracted from parasitoid larvae followed by restriction endonuclease digestion of PCR products to distinguish P. howardi from other potentially co-occurring Peristenus species. Peak parasitism of Lygus nymphs occurred between the first and third weeks of July for both years for all host plants sampled. Of the parasitoid larvae recovered from Lygus nymphs in our study, 75% to 80% tested positive as Peristenus spp. and 76% of these matched the endonuclease digestion banding pattern for P. howardi. The identity of the remaining 20% to 25% of the parasitoids is not known. PMID- 17119627 TI - Molecular cloning and nucleotide sequence of CYP6BF1 from the diamondback moth, Plutella xylostella. AB - A novel cDNA clong encoding a cytochrome P450 was screened from the insecticide susceptible strain of Plutella xylostella (L.) (Lepidoptera:Yponomeutidae). The nucleotide sequence of the clone, designated CYP6BF1, was determined. This is the first full-length sequence of the CYP6 family from Plutella xylostella (L.). The cDNA is 1661bp in length and contains an open reading frame from base pairs 26 to 1570, encoding a protein of 514 amino acid residues. It is similar to the other insect P450s in gene family 6, including CYP6AE1 from Depressaria pastinacella, (46%). The GenBank accession number is AY971374. PMID- 17119628 TI - Host-parasitoid interactions relating to penetration of the whitefly, Bemisia tabaci, by the parasitoid wasp, Eretmocerus mundus. AB - It has been reported that the aphelinid wasp Eertmocerus mundus parasitizes all four nymphal instars of the sweet potato whitefly, Bemisia tabaci (Biotype B), with 3rd instars being the preferred hosts. The parasitoid lays its egg on the leaf underneath the host nymph. First instars hatch and later penetrate the whitefly. Previous studies have shown that the initiation of parasitoid penetration induces the host to form a cellular capsule around the parasitoid. As described here, females never oviposited once the 4th instar whitefly nymph had initiated adult development. First instar E. mundus larvae were observed under 2nd, 3rd and 4th instar whitefly nymphs, however, penetration did not occur until the whitefly had reached the 4th instar. The non-penetrating E. mundus larva almost always induced permanent developmental arrest in its 4th instar whitefly host and also caused a reduction in whole body host ecdysteroid titers. Therefore, unless there is a peak in molting hormone titer in the area local to penetration, it appears that the induction of capsule formation is not due to an increase in ecdysteroid titer. As the capsule formed around the penetrating parasitoid, host epidermal cells multiplied and became cuboidal and columnar, and relatively thick layers of new cuticle were deposited within the developing capsule, particularly near its ventral opening. The newly formed host cuticle was thinner in the dorsal part of the capsule and appeared to be absent at its apex. These results provide new information regarding the timing and dynamics of parasitoid oviposition and egg hatch as related to larval penetration, parasitoid induced changes in whitefly development, molting hormone titers and the process of capsule formation. PMID- 17119629 TI - Cross-species investigation of Helicoverpa armigera microsatellites as potential markers for other related species in the Helicoverpa--Heliothis complex. AB - Primers previously designed to amplify microsatellite DNA markers in the Old World bollworm, Helicoverpa armigera, larvae were tested in three closely related species: the corn earworm, Helicoverpa zea, tobacco budworm, Heliothis virescens, and Heliothis subflexa. Of the fourteen loci surveyed, only four loci (HaB60, HaC14, HaC87, HarSSR1) consistently demonstrated scorable single-copy microsatellite bands. Of these four, length polymorphism was identified only in the HaB60 marker (160 bp, 140 bp) of the H. virescens and H. subflexa sampled laboratory populations. Partial DNA sequences of all the identified single-copy microsatellites are presented as well as alignments to their respective H. armigera microsatellite. PMID- 17119630 TI - Influence of diet on the larval beet armyworm, Spodoptera exigua, glucose oxidase activity. AB - Many researchers investigating plant-insect interactions maintain their insect colonies on artificial diet and assume that salivary enzymes and elicitors remain representative of natural situations. These salivary elicitors, such as the enzyme glucose oxidase (GOX), play important roles in influencing plant defense responses. In fact, GOX has been implicated in suppressing induced nicotine production in tobacco plants (Musser et al., 2002). In this study, we investigated the effect of artificial or plant diet on the GOX activity in caterpillars of the beet armyworm, Spodoptera exigua. In the later developmental stages, whole body GOX of S. exigua caterpillars reared on a wheat germ-based artificial diet is over ten times higher than when insects were fed plants of the legume, Medicago trunctula. Labial salivary GOX accounted for most of this whole body activity in 4th instar caterpillars (57.8%), with the remaining activity present in the carcass. Hemolymph GOX levels were below the detection limits of the o-dianisidine-peroxidase assay used to measure activity. Labial salivary GOX activity was significantly higher in 4th larval instars reared on artificial diet compared with plant-fed caterpillars (U/mg per pair labial salivary gland, p = 0.0062), suggesting that diet effects GOX activity. When 4th instar plant-fed caterpillars were transferred to artificial diet, increased labial salivary GOX activity is closely correlated with the amount of time spent feeding on artificial diet. This study shows that the labial salivary GOX activity of S. exigua caterpillars is dependent on diet and developmental stage and that caution must be exercised in the design of plant-insect experiments. PMID- 17119631 TI - Impact of riparian land use on stream insects of Kudremukh National Park, Karnataka state, India. AB - The impact of riparian land use on the stream insect communities was studied at Kudremukh National Park located within Western Ghats, a tropical biodiversity hotspot in India. The diversity and community composition of stream insects varied across streams with different riparian land use types. The rarefied family and generic richness was highest in streams with natural semi evergreen forests as riparian vegetation. However, when the streams had human habitations and areca nut plantations as riparian land use type, the rarefied richness was higher than that of streams with natural evergreen forests and grasslands. The streams with scrub lands and iron ore mining as the riparian land use had the lowest rarefied richness. Within a landscape, the streams with the natural riparian vegetation had similar community composition. However, streams with natural grasslands as the riparian vegetation, had low diversity and the community composition was similar to those of paddy fields. We discuss how stream insect assemblages differ due to varied riparian land use patterns, reflecting fundamental alterations in the functioning of stream ecosystems. This understanding is vital to conserve, manage and restore tropical riverine ecosystems. PMID- 17119632 TI - Susceptibility of biological stages of the horn fly, Haematobia irritans, to entomopathogenic fungi (Hyphomycetes). AB - The susceptibility of the egg, pupa, and adult of Haematobia irritans (L.) (Diptera: Muscidae) to isolates of the fungi Metarhizium anisopliae (Metsch.) Sor., Beauveria bassiana (Bals.) Vuill., and Paecilomyces fumosoroseus (Wize) Brown and Smith, was evaluated under laboratory conditions. Groups of 20 eggs than 4 h old, pupae less than 48h old and adults were sprayed with a conidial suspension of each isolate. Eggs, pupae and adults of horn fly were susceptible to these entomopathogenic fungi. For treated eggs, the isolates Ma3, Ma 15, Ma25, Pfr1, and Pfr8 reduced adult emergence to 3.8% to 6.3% in comparison with the control (72%). The mortality of pupae infected by the isolates Ma2, Ma25, and Pfr10 ranged between 50% and 71.3%. Mortality of adults after treatment with the isolates Ma6, Ma 10, Ma 14, Ma 15, Pfr 1, Pfr 9, Pfr 10, Pfr 11, and Pfr12 were higher than 90%. The isolate Ma6 produced the lowest LC(50) against adult horn flies (8.08 × 10(2)conidia/ml). These findings supported the hypotheses that isolates of M. anisopliae, and P. fumosoroseus are pathogenic against the different biological stages of horn flies by reducing adult emergence when applied on groups of eggs and pupae, and producing mortality when applied to adults. PMID- 17119634 TI - Bacterial pathways for degradation of nitroaromatics. AB - The last one hundred years have seen a massive expansion in the chemicals industry; however, with this progress came the concomitant pollution of the environment with a significant range of xenobiotics.Nitroaromatic compounds form one such category of novel environmental contaminants and are produced through a large number of industrial processes, most notably the pesticides, dyes and explosives industries. Whilst singly nitrated aromatic compounds are usually mineralised in the environment, multiply nitrated aromatics, such as the explosive 2,4,6-trinitrotoluene (TNT), are recalcitrant and highly toxic. The predominant route of biological transformation of aromatic compounds is oxidation; however, the presence of three electron-withdrawing nitro-groups around the ring prevents oxidation, rendering such compounds resistant to biodegradation. The subsequent accumulation of these contaminants has stimulated much research leading to the isolation of bacteria that possess, to varying extents, the ability to remediate explosives and other nitroaromatic pollutants.The extreme environments created by these toxic substances accelerate the evolutionary process and examples of bacteria that have conscripted metabolic enzymes for novel remediatory pathways are included. This Highlight ends with a discussion of the future of nitroaromatic bioremediation including engineering plants to express bacterial enzymes for use in bioremediation programs. PMID- 17119635 TI - A scent of therapy: pharmacological implications of natural products containing redox-active sulfur atoms. AB - A range of sulfur-containing natural products from plants, fungi, bacteria and animals have recently been investigated to determine their therapeutic potential. Preliminary in vitro and in vivo studies of compounds such as ergothioneine, ovothiols, allicin, leinamycin, varacin, lenthionine and diallyltetrasulfide have provided evidence for antioxidant, antibacterial, antimicrobial, antifungal and anticancer properties. The biological activity of these compounds is the result of specific chemical properties which converge in chemotypes such as thiols, disulfides, sulfenic and sulfinic acids,thiosulfinates, sulfoxides, sulfones and polysulfides. Redox-activity, catalysis, metal binding, enzyme inhibition and radical generation allow reactive sulfur species to interact with oxidative stressors, to affect the function of redox-sensitive cysteine proteins and to disrupt the integrity of DNA and cellular membranes. In some cases, the biological activity of sulfur-containing plant products depends on initial enzymatic activation, which allows thiosulfinates and isothiocyanates to be generated with high target selectivity. Not surprisingly, research into the biochemical and pharmacological properties of the lesser known sulfur chemotypes is rapidly gathering momentum. PMID- 17119636 TI - Biosynthesis of 2-deoxystreptamine-containing aminoglycoside antibiotics. AB - The 2-deoxystreptamine-containing aminoglycosides are an important class of clinically valuable antibiotics. A deep understanding of the biosynthesis of these natural products is required to enable efforts to rationally manipulate and engineer the biological production of novel aminoglycosides. This review discusses the development of our biosynthetic knowledge over the past half century, with emphasis on the relatively recent contributions of molecular biology to the elucidation of these biosynthetic pathways. PMID- 17119637 TI - Diterpenoids. AB - This review covers the isolation and structures of diterpenoids, including labdanes, clerodanes,pimaranes, abietanes, kauranes, cembranolides, taxanes and marine diterpenoids. The literature from January to December 2005 is reviewed, and 195 references are cited. PMID- 17119638 TI - Steroids: partial synthesis in medicinal chemistry. AB - This article reviews the progress in the chemistry of the steroids that was published between January and December 2005. The reactions and partial synthesis of estrogens, androgens, pregnanes, bile acid derivatives, cholestanes and vitamin D analogues are covered. There are 139 references. PMID- 17119639 TI - Dissecting non-ribosomal and polyketide biosynthetic machineries using electrospray ionization Fourier-Transform mass spectrometry. AB - Many virulence factors and bioactive compounds with antifungal, antimicrobial, and antitumor properties are produced via the non-ribosomal peptide synthetase (NRPS) or polyketide synthase(PKS) paradigm. During the biosynthesis of these natural products, substrates, intermediates and side products are covalently tethered to the NRPS or PKS catalyst, introducing mass changes, making these biosynthetic systems ideal candidates for interrogation by large molecule mass spectrometry. This review serves as an introduction into the application of electrospray ionization Fourier-Transform massspectrometry (ESI-FTMS) to investigate NRPS and PKS systems. ESI-FTMS can be used to understand substrate tolerance, timing of covalent linkages, timing of tailoring reactions and the transfer of substrates and biosynthetic intermediates from domain to domain. Therefore we not only highlight key mechanistic insights for thiotemplate systems as found on the enterobactin,yersiniabactin, epothilone, clorobiocin, coumermycin, pyoluteorin, gramicidin, mycosubtilin, C-1027,6-deoxyerythronolide B and FK520 biosynthetic pathways, but we also explain the approaches taken to identify active sites from complex digests and compare the FTMS based assay to traditional assays and other mass spectrometric techniques. Although mass spectrometry was introduced over two decades ago to investigate NRPS and PKS biosynthetic systems, this is the first review devoted to this methodology. PMID- 17119640 TI - Birch bark research and development. AB - This review will detail progress made in the previous decade on the chemistry and bioactivity of birch bark extractive products. Current and future applications of birch bark natural products in pharmaceuticals, cosmetics, and dietary supplements for the prevention and treatment of cancer, HIV,and other human pathogens are reviewed. Current developments in the technology of birch bark processing are discussed. New approaches for the synthesis of potentially valuable birch bark triterpenoid derivatives are also reviewed. PMID- 17119641 TI - Natural sesquiterpenoids. AB - This review covers the isolation, structural determination, synthesis and chemical and microbiological transformations of natural sesquiterpenoids. The literature from January to December 2005 is reviewed,and 386 references are cited. PMID- 17119642 TI - Electron transfer processes in DNA: mechanisms, biological relevance and applications in DNA analytics. AB - In principle, DNA-mediated charge transfer processes can be categorized as oxidative hole transfer and reductive electron transfer. With respect to the routes of DNA damage most of the past research has been focused on the investigation of oxidative hole transfer or transport. On the other hand, the transport or transfer of excess electrons has a large potential for biomedical applications, mainly for DNA chip technology. PMID- 17119643 TI - Indolocarbazole natural products: occurrence, biosynthesis, and biological activity. AB - The indolocarbazole family of natural products, including the biosynthetically related bisindolylmaleimides, is reviewed (with 316 references cited). The isolation of indolocarbazoles from natural sources and the biosynthesis of this class of compounds are thoroughly reviewed, including recent developments in molecular genetics, enzymology and metabolic engineering. The biological activities and underlying modes of action displayed by natural and synthetic indolocarbazoles is also presented, with an emphasis on the development of analogs that have entered clinical trials for its future use against cancer or other diseases. PMID- 17119644 TI - From miso, sake and shoyu to cosmetics: a century of science for kojic acid. AB - This article reviews the curious history of kojic acid, discovered as a fungal natural product in 1907. It was one of the first secondary metabolites to have its biosynthetic pathway studied by the isotope tracer technique, and, more recently, has been of interest as a skin lightening agent. There are 112 references. PMID- 17119645 TI - Calculations of solute and solvent entropies from molecular dynamics simulations. AB - The translational, rotational and conformational (vibrational) entropy contributions to ligand-receptor binding free energies are analyzed within the standard formulation of statistical thermodynamics. It is shown that the partitioning of the binding entropy into different components is to some extent arbitrary, but an appropriate method to calculate both translational and rotational entropy contributions to noncovalent association is by estimating the configurational volumes of the ligand in the bound and free states. Different approaches to calculating solute entropies using free energy perturbation calculations, configurational volumes based on root-mean-square fluctuations and covariance matrix based quasiharmonic analysis are illustrated for some simple molecular systems. Numerical examples for the different contributions demonstrate that theoretically derived results are well reproduced by the approximations. Calculation of solvent entropies, either using total potential energy averages or van't Hoff plots, are carried out for the case of ion solvation in water. Although convergence problems will persist for large and complex simulation systems, good agreement with experiment is obtained here for relative and absolute ion hydration entropies. We also outline how solvent and solute entropic contributions are taken into account in empirical binding free energy calculations using the linear interaction energy method. In particular it is shown that empirical scaling of the nonpolar intermolecular ligand interaction energy effectively takes into account size dependent contributions to the binding free energy. PMID- 17119646 TI - Molecular simulation studies of water physisorption in zeolites. AB - We report a series of Grand Canonical Monte Carlo simulations of water adsorption in NaY and NaX faujasite, as well as in silicalite-1. Computed adsorption isotherms and heats of adsorption were in good agreement with the available experiments. The existence of cyclic water hexamers in NaX located in the 12-ring windows, recently disclosed by neutron diffraction experiments (Hunger et al., J. Phys. Chem. B, 2006, 110, 342-353) was reproduced in our simulations. Interestingly enough, such cyclic hexamer clusters were also observed in the case of NaY, in which no stabilizing cation is present in the 12-ring window. We also report cation redistribution upon water adsorption for sodium faujasite with varying cation contents (Si ratio Al ratio in the range 1.53-3). A simple and transferable forcefield was used, that enabled to reproduce the different aspects of water physisorption in stable zeolites. The high pressure water condensation in hydrophobic silicalite-1 was reproduced without any parameter readjustment. The method and forcefield used here should be useful for engineering oriented applications such as the prediction of multi-component mixture adsorptive separations in various stable zeolites. It allows to address the issue of the effect of the small amounts of water that are almost inevitably present in zeolite-based separation processes. PMID- 17119647 TI - A hollow tetrahedral cage of hexadecagold dianion provides a robust backbone for a tuneable sub-nanometer oxidation and reduction agent via endohedral doping. AB - We show, via density functional theory calculations, that dianionic Au16(2-) cluster has a stable, hollow, Td symmetric cage structure, stabilized by 18 delocalized valence electrons. The cage maintains its robust geometry, with a minor Jahn-Teller deformation, over several charge states (q = -1,0,+1), forming spin doublet, triplet and quadruplet states according to the Hund's rules. Endohedral doping of the Au16 cage by Al or Si yields a geometrically robust, tuneable oxidation and reduction agent. Si@Au16 is a magic species with 20 delocalized electrons. We calculate a significant binding energy for the anionic Si@Au16/O2- complex and show that the adsorbed O2 is activated to a superoxo species, a result which is at variance with the experimentally well-documented inertness of Au16- anion towards oxygen uptake. PMID- 17119648 TI - A microwave-heated infrared reaction cell for the in situ study of heterogeneous catalysts. AB - A transmission infrared microreactor cell which holds a pressed disc in a controlled atmosphere and allows microwave and conventional heating up to 423 K is demonstrated using the oxidation of carbon monoxide over the standard catalyst EUROPT-1. Optical characteristics are determined by the choice of CaF2 as the window material, allowing transmission from 77,000-1000 cm(-1). An oscillating microwave power regime with a peak height of 200 W is used and time-resolved infrared spectra and mass spectrometry show oscillations in the reaction which correspond to the microwave heating. PMID- 17119649 TI - Size effects of gold nanaoparticles on plasmon-induced photocurrents of gold-TiO2 nanocomposites. AB - An ITO electrode was coated with a nanoporous TiO2 film, and the film was loaded with Au nanoparticles with different diameters (15, 40 and 100 nm). The electrode exhibited plasmon-induced photocurrents in the presence of Fe(2+/3+) couple under visible light (lambda > 420 nm). The quantum efficiency increased with increasing particle size, whereas the maximum photocurrent density decreased. PMID- 17119650 TI - Nonequilibrium thermodynamics--A tool to describe heterogeneous catalysis. AB - In the study of multi-component mass transfer it is common to use the film model, in which all the resistance to mass transfer towards a catalytic surface is assumed to be localized in a diffusion layer in front of the surface. At the surface one furthermore assumes that the temperature and chemical potentials are continuous, while the coupling of a possible heat flux to the mass fluxes is assumed to be negligible. Both these assumptions are questionable. Using nonequilibrium thermodynamics we discuss how to integrate the coupling between heat and mass fluxes in the description of the film. Furthermore, following Gibbs, we introduce the surface as a separate thermodynamic system where the coupling between the vectorial heat flux and the scalar reaction rate is allowed and can be significant in heterogeneous catalysis. Non-equilibrium thermodynamic theory for surfaces allows one to find the proper rate equations. It allows for a consistent and complete description of mass and heat transfer through the film and subsequently from the film to the surface where the reaction takes place. Fast endo- or exothermic surface reactions in heterogeneous catalysis may give significant temperature gradients between a catalyst surface and the media, which will, when not accounted for, lead to an incorrect evaluation of the activity, stability and selectivity of a catalyst. Non-equilibrium thermodynamics is a useful tool for predicting the surface temperature as well as for analyzing the system. In this contribution we sketch how to systematically set up the complete description, in which the film and the surface "sum up" to one effective surface. PMID- 17119651 TI - An ab initio investigation of zinc chloro complexes. AB - A series of geometry, frequency, and energy calculations of chloroaquazinc(II) complexes were carried out at up to the MP2/6-31+G* level. A thorough examination of all species up to and including hexacoordinate species, and with up to six chlorides, was carried out. The structures of the complexes are compared with experimental data where available. The solution chemistry of zinc(II) in the presence of chloride is discussed, and Raman spectra of zinc perchlorate with increasing amount of chloride are presented. PMID- 17119652 TI - Electrochemical properties of core-shell TiC-TiO2 nanoparticle films immobilized at ITO electrode surfaces. AB - Titanium carbide (TiC) nanoparticles are readily deposited onto tin-doped indium oxide (ITO) electrodes in the form of thin porous films. The nanoparticle deposits are electrically highly conducting and electrochemically active. In aqueous media (at pH 7) and at applied potentials positive of 0.3 V vs. SCE partial anodic surface oxidation and formation (at least in part) of novel core shell TiC-TiO2 nanoparticles is observed. Significant thermal oxidation of TiC nanoparticles by heating in air occurs at a temperature of 250 degrees C and leads first to core-shell TiC-TiO2 nanoparticles, next at ca. 350 degrees C to TiO2 (anatase), and finally at temperatures higher than 750 degrees C to TiO2 (rutile). Electrochemically and thermally partially oxidized TiC nanoparticles still remain very active and for some redox systems electrocatalytically active. Scanning and transmission electron microscopy (SEM and TEM), temperature dependent XRD, quartz crystal microbalance, and voltammetric measurements are reported. The electrocatalytic properties of the core-shell TiC-TiO2 nanoparticulate films are surveyed for the oxidation of hydroquinone, ascorbic acid, and dopamine in aqueous buffer media. In TiC-TiO2 core-shell nanoparticle films TiO2 surface reactivity can be combined with TiC conductivity. PMID- 17119653 TI - Effects of noncovalently bound quinones on the ground and triplet states of zinc chlorins in solution and bound to de novo synthesized peptides. AB - The Qy absorption band of two chlorophyll derivatives, zinc chlorin e6 (ZnCe6) and zinc pheophorbide a (ZnPheida), in aqueous solution is bathochromically shifted on addition of quinones, e.g., 1,4-benzoquinone (BQ), with a corresponding shift of the fluorescence band. This is due to a complex formation of zinc chlorins induced by BQs and subsequent rearrangement. The time-resolved absorption spectra after laser pulse excitation show triplet quenching of the pigments by BQ and other quinones via electron transfer. The effects of electron transfer to noncovalently bound BQs were also studied with de novo synthesized peptides, into which ZnCe6 and ZnPheida were incorporated as model systems for the primary steps of photosynthetic reaction centers. Whereas the photophysical properties are similar to those of the unbound zinc chlorins, no BQ-mediated complex formation was observed. PMID- 17119654 TI - A crossed beams study of the reaction of carbon atoms, C(3Pj), with vinyl cyanide, C2H3CN(X 1A')--investigating the formation of cyano propargyl radicals. AB - The chemical dynamics of the reaction of ground state carbon atoms, C(3Pj), with vinyl cyanide, C2H3CN(X 1A'), were examined under single collision conditions at collision energies of 29.9 and 43.9 kJ mol(-1) using the crossed molecular beams approach. The experimental studies were combined with electronic structure calculations on the triplet C4H3N potential energy surface (H. F. Su, R. I. Kaiser, A. H. H. Chang, J. Chem. Phys., 2005, 122, 074320). Our investigations suggest that the reaction follows indirect scattering dynamics via addition of the carbon atom to the carbon-carbon double bond of the vinyl cyanide molecule yielding a cyano cyclopropylidene collision complex. The latter undergoes ring opening to form cis/trans triplet cyano allene which fragments predominantly to the 1-cyano propargyl radical via tight exit transition states; the 3-cyano propargyl isomer was inferred to be formed at least a factor of two less; also, no molecular hydrogen elimination channel was observed experimentally. These results are in agreement with the computational studies predicting solely the existence of a carbon versus hydrogen atom exchange pathway and the dominance of the 1-cyano propargyl radical product. The discovery of the cyano propargyl radical in the reaction of atomic carbon with vinyl cyanide under single collision conditions implies that this molecule can be an important reaction intermediate in combustion flames and also in extraterrestrial environments (cold molecular clouds, circumstellar envelopes of carbon stars) which could lead to the formation of cyano benzene (C6H5CN) upon reaction with a propargyl radical. PMID- 17119655 TI - Increased layer interdiffusion in polyelectrolyte films upon annealing in water and aqueous salt solutions. AB - As-deposited films of multilayered polyelectrolytes are considered to be non equilibrium structures. Due to the strong attraction between oppositely charged polyions, polyelectrolyte interdiffusion is thought to be suppressed during the adsorption process. Equilibration is promoted by a decrease of the electrostatic attraction between polyion pairs. We have used neutral impact collision ion scattering spectroscopy to investigate the influence of polyelectrolyte multilayer annealing in water and aqueous 1 M NaCl solutions at different temperatures (20 and 70 degrees C) on the increase in interpenetration of a single polyelectrolyte layer throughout the whole film. The multilayers were composed of poly(4-vinylpyridinium) and poly(4-styrenesulfonate). Contrast between neighboring layers was established by labelling the layer in question with the heavy atom ruthenium. It is found that both temperature and salt increase layer interpenetration, whereas salt has a stronger influence than temperature. From numerical simulations polyelectrolyte diffusion coefficients were evaluated for the different annealing conditions. The influence of temperature and salt on the equilibration of the film is interpreted in terms of increased screening of polyion charges and binding of small counterions to polyion monomeric units. PMID- 17119656 TI - Hepatitis C: facts in numbers. PMID- 17119657 TI - [Ambulatorial prevalence of hepatitis B and C markers in patients with human immunodeficiency virus infection in a general hospital]. AB - BACKGROUND: Hepatitis B and C viruses and human immunodeficiency virus share the same route of transmission, and the prevalence of HBV and HCV infection in patients infected with HIV is greater than it is in the general population. AIM: To determine the prevalence of hepatitis B and C markers in a population of patients with HIV as well as the risk factors involved. PATIENTS AND METHODS: From 5,870 registration forms of patients with HIV of an Infectology Unit, 587 were randomly selected. From these, the 343 which had investigated the presence of any hepatitis B (HBsAg, anti-HBc or anti-HBs) or C (anti-HCV) marker were retrospectively analyzed. RESULTS: HBsAg was positive in 14/306 (4.6%), anti-HBs was positive in 40/154(26.0%), and anti-HBc in 79/205 (38.5%). The anti-HCV test was reactive in 126/330 (38.2%). HBV and HCV co-infection was observed in 7 of the 296 patients who had both HBsAg and anti-HCV tests (2.4%). For those who were HBsAg positive, the main exposure factor was homosexual intercourse (50.0%). For those who were anti-HCV reactive, the main risk factor was intravenous drug use (75.3%). In the HIV mono-infected (185 patients), the most prevalent exposure risk factor was promiscuous heterosexual practices or sexual intercourse with a spouse infected with HIV (83 patients - 44.9%). CONCLUSION: In our environment HBV-HIV and HCV-HIV co-infections are frequent, a greater relevance being observed in the association between HCV and HIV. PMID- 17119658 TI - [Prevalence of hepatitis C infection in patients with type 2 diabetes mellitus]. AB - BACKGROUND: Recently, a possible epidemiological association between hepatitis C virus infection and diabetes mellitus has been suggested and a higher prevalence of HCV antibodies has been found among type 2 diabetic when compared with normal controls. AIM: To evaluate the prevalence of hepatitis C infection in diabetic patients in Curitiba, PR, Brazil. PATIENTS AND METHODS: A total of 145 type 2 and 104 type 1 diabetic patients attending the outpatient diabetic unit of an university hospital were consecutively tested for anti-HCV, using a fourth generation enzyme-linked immunosorbent assay (ELISA). The control group was constituted by 16,720 volunteer blood donors attending the blood bank of the same hospital during the period of the study. Diabetic patients were also evaluated for clinical, biochemical (aminotransferase levels) and demographic variables and previous exposure to risk factors for hepatitis C infection. RESULTS: A higher prevalence of hepatitis C infection was observed in type 2 diabetic patients in comparison with blood donors. Although anti-HCV prevalence in type 2 diabetic patients was higher than found in type 1, it did not reach statistical significance. Both diabetic groups were predominantly female, and as expected, type 2 diabetic were older than type 1. Race distribution, duration of the disease, and previous exposure to hepatitis C risk factors were similar in both groups, but type 2 diabetic subjects had higher median levels of alanine aminotransferase than type 1. CONCLUSIONS: A higher prevalence of hepatitis C infection was detected in type 2 diabetic patients in comparison with blood donors in our region, in accordance with study data from different populations. If all type 2 diabetic patients should undergo regular screening for hepatitis C infection remains a question. PMID- 17119659 TI - Prevalence of hepatitis C virus in alcoholic patients: role of parenteral risk factors. AB - BACKGROUND: The prevalence of hepatitis C virus (HCV) infection is elevated in alcoholic patients, but the risk factors are unclear. The role of parenteral risk factors are indeterminated in this population. AIMS: To determine the prevalence of hepatitis C virus infection in alcoholic patients admitted to a detoxification unit and to evaluate the presence of underlying parenteral risk factors. METHODS: A total of 114 consecutive unselected alcoholic patients admitted to a single chemical dependency unit during 14 month were included. Epidemiological data and history of parenteral risk factors for hepatitis C virus infection were obtained with a standardized questionnaire. Blood was collected for determination of aminotransferases and anti-hepatitis C virus antibodies (ELISA-3). Positive samples were confirmed by polymerase chain reaction and tested for genotype. RESULTS: Among the 114 alcoholics, 17 (15%) were anti-hepatitis C virus positive. Of these, 12 (71%) had detectable serum HCV-RNA by PCR. Genotype 1 was found in six cases and genotype 3 in five (one patient was undetermined). Forty-nine (43%) patients had elevated serum ALT and/or AST at baseline. The comparison between the 17 positive and the 97 negative patients showed significant differences in mean serum ALT levels (42 +/- 41 IU/L vs. 22 +/- 20 IU/L), rate of elevated ALT (65% vs. 34%), and presence of parenteral risk factors (94% vs. 10%). Comparison between alcoholic patients with and without elevated aminotransferases showed significant difference only in the rate of positive anti-hepatitis C virus antibodies (24% vs. 7%). Furthermore, among the 17 anti-hepatitis C virus positive patients, the rate of detectable HCV-RNA was significantly higher in the 12 with elevated aminotransferases versus the 5 with normal aminotransferases (92% vs. 20%). CONCLUSIONS: There was a high prevalence of anti-hepatitis C virus antibodies in alcoholics and the majority was confirmed by the presence of detectable HCV-RNA. Intravenous drug use was the main risk factor for hepatitis C virus infection in this population. PMID- 17119660 TI - [Lymphoma in patients harboring hepatosplenic mansonic schistosomiasis]. AB - BACKGROUND: Correlation between infectious agents and linfoproliferative diseases are more stablished, over all virus and bacteria, through the activation of linfocytes. AIM: To describe six new cases, of a series of 254 patients (2,36%) with mansonic schistosomiasis, in the hepatosplenic form. METHODS: Six patients will be described, amongst the 254 carriers of mansonic schistosomiasis, in the hepatosplenic form, followed in the last 13 years. RESULTS: All the six cases had occurred in women. The histopathologic examinations had evidenced two cases of marginal splenic zone lymphomas, one of great cells with immunoblasts lymphomas, one diffuse lymphomas, a great cells malignant lymphomas, a great not clivads cells, and another case of Hodgkin. Half of the six evolved for the death 4-15 months after the diagnosis. The others three persist in accompaniment in the Oncology Division of the Clinics Hospital. CONCLUSION: The incidence of lymphomas in the 254 mansonic schistosomiasis patients followed in our clinic was of 2,36%. At last, this article intends to call the attention, for the occurrence of lymphomas, in the spleen of patients with mansonic schistosomiasis, in the hepatosplenic form. PMID- 17119661 TI - [Lymph node evaluation in colorectal cancer]. AB - BACKGROUND: In Brazil, colorectal carcinoma is the third cause of death by malignant tumors among women, and the fifth among men. Lymph node involvement is one of the most relevant prognostic maker in these tumors. AIM: To study lymph node involvement in colorectal carcinoma in relationship to biological behavior and tumor location. PATIENTS AND METHODS: One hundred and eight five colorectal carcinoma cases were studied. Lymph node involvement was analyzed according to tumor location, diameter, vessel invasion, and TNM staging. RESULTS: Three thousand nine hundred and six lymph nodes were harvested in 185 patients (21.1 lymph nodes/patient). Metastasis were detected in 399/2,573 peritumoral lymph nodes (15.5%) and in 72/1,333 non-peritumoral lymph nodes (5.4%). Eighty-six patients presented metastasis; in these patients 471/1942 lymph nodes were compromised. In 26 patients peritumoral and non-peritumoral lymph nodes were involved; in 57 cases metastasis were detected only in peritumoral lymph nodes and in 3, only non-peritumoral lymph nodes were involved. The number of lymph node was higher among cecal tumors and smaller in the rectum and sigmoid. There was a positive correlation between the number of metastatic lymph node and pT, tumor diameter and lymphatic and venous invasion; there was a negative correlation between lymph node involvement and lymphocytic response; pN was significantly associated with pT. CONCLUSIONS: Colorectal carcinoma involves preferentially peritumoral lymph node, but in 29 patients (15,7%) non-peritumoral lymph nodes were affected, which is important for tumor staging and prognosis. pN and the number of metastatic lymph nodes were associated with other behaviour markers. PMID- 17119662 TI - [Evaluation of the toxicity and quality of life in patients with colorectal cancer treated with chemotherapy]. AB - BACKGROUND: The colorectal cancer is the fourth cause of cancer in Brazil and 5 fluorouracil is the drug most commonly used in the adjuvant or palliative treatment of this disease. AIM - Evaluating in patients with colorectal cancer and chemotherapy, the toxicity and the quality of life. PATIENTS AND METHODS: From March 2001 and May 2003, 45 patients treated with colorectal cancer treated with 5-fluourouracil and folinic acid were followed closely during six cycles. The gastrointestinal and hematologic toxicity was analysed making use of the chart "Recommendations for the Graduation of Acute and Subacute Toxicity". After the end of each cycle of chemotherapy, the results were registered according to the respectives degrees that vary from 0 to 4. The quality of life was researched through the WHOQOL bref (World Health Organization Quality of Life) questionary that consists of 26 questions and 4 domains: physical, psychological, social relations and environmental, in the beginning, on the 3rd and 6th cycles of treatment. RESULTS: Among the 45 patients, 28 were male, the average age was 58.4 years old (from 34 to 79 years old). According to the International Union Against Cancer classification, 34 patients (75.6%) had tumors stage II or III and 11 had tumors stage IV (24.4%), 64.4% were in the colon. In 57.7% the chemotherapy was adjuvant and in the others palliative. The toxicities more commonly found were nauseas (42%), diarrhea (38%), and neutropenia (15.7%). There was no significant difference among the degrees of toxicity in the different cycles as well as among the patients in adjuvant or palliative treatment. Significant alterations was found among the quality of life in the physical and psychological domains when the 1st and the 2nd or the 1st and the 3rd application of the test were done. Alterations of the quality of life were also found in the social domain when the first evaluation was compared with the last one. There was no difference between the quality of life and the treatment. PMID- 17119663 TI - [Fecal incontinence in postmenopausal women: prevalence, severity and associated factors]. AB - BACKGROUND: Fecal incontinence occurs more frequently in the female population and it becomes more prevalent with increasing age. There are few studies that have assessed the prevalence and severity of postmenopausal women. AIMS: To investigate fecal incontinence. PATIENTS AND METHODS: A cross-sectional study was performed on 100 postmenopausal women over the age of 45. Sociodemographic and clinical characteristics were evaluated, and a descriptive analysis of these characteristics was carried out. The prevalence of fecal incontinence was estimated. St. Mark's incontinence score was applied to study the severity of symptoms associated with fecal incontinence. The score was then categorized according to the tertile and symptom severity was classified as mild, moderate or severe incontinence. Bivariate and multivariate analyses were used to study the association between fecal incontinence and its likely determinants, employing the prevalence ratio. Confidence interval was set at 95%. RESULTS: The mean age of the patients was 58.9 +/- 5.9 years (range, 46-76 years). The prevalence rate was 15% for fecal incontinence. Of incontinent patients, 60% had mild incontinence. After multivariate analysis, factors associated with fecal incontinence was history of forceps delivery (prevalence ratio: 7.80; 95% confidence interval:2.38 25.55). CONCLUSIONS: The prevalence of fecal incontinence was high in postmenopausal women. Data suggest that most women presented mild fecal incontinence. The history of forceps delivery was associated with fecal incontinence. PMID- 17119664 TI - [Aging and esophageal motility in patients with gastroesophageal reflux disease]. AB - BACKGROUND: Alterations of esophageal contractions may worsen the esophageal lesions caused by gastroesophageal reflux. The impairment of the contractions may be localized only in the distal esophagus or in the entire esophageal body, and may be worse with the aging process. AIMS: To evaluate the proximal and distal esophageal contractions in patients with gastroesophageal reflux symptoms with or without esophagitis. PATIENTS AND METHODS: We studied esophageal motility in 104 patients with gastroesophageal reflux symptoms, 42 with normal esophageal endoscopic examination, 47 with mild esophagitis and 15 with severe esophagitis. The esophageal contractions were measured by the manometric method at 2, 7, 12 and 17 cm from the upper esophageal sphincter, after five swallows of a 5 mL bolus of water. RESULTS: The amplitude and area under the curve of contractions were lower in patients with severe esophagitis than in patients without esophagitis or with mild esophagitis in the distal part of the esophageal body (17 cm from the upper esophageal sphincter). In the proximal esophageal body there was no difference in amplitude or area under the curve. In the entire esophageal body there was no difference between the three groups of patients in duration, velocity of peristaltic contractions, or proportion of failed, simultaneous, non-propagated or peristaltic contractions. There was no difference between the patients with less than 50 years or with more than 50 years of age. CONCLUSIONS: Patients with severe esophagitis had lower distal esophageal contraction amplitude than patients without esophagitis or with moderate esophagitis. There was no effect of aging on esophageal contractions. PMID- 17119665 TI - [Electromanometry and acid pHmetry of 24 hours in postoperative evaluation of the hiatoplasty and total antireflux wrap laparoscopic]. AB - BACKGROUND: The electromanometry and 24-hour esophageal pHmetry has been proposed as objective method in postoperative evaluation of antireflux surgery. AIM: Prospective evaluation of the results of cruroplasty and total fundoplication in the treatment of non-complicated gastroesophageal reflux disease based on comparative results of pre and post operative manometry and pHmetry findings. METHODS: Fifty-nine consecutive individuals with typical symptoms of gastroesophageal reflux disease were prospectively studied. All patients were submitted to a laparoscopic short floppy Nissen fundoplication between March, 2002 and August, 2003. All patients were submitted pre and postoperatively (3 months) to upper digestive endoscopy, manometry and pHmetry 24 hours. RESULTS: Eighteen (30.5%) patients were male and 41 (69.5%) female, their average age was 43.8 years. There were differences comparing the pre and post operative period in the manometrical findings in the following items: localization of the lower esophageal sphincter (41.2 cm and 42.3 cm). Extension o f the lower esophageal sphincter preoperative (2.0 cm and 2.5 cm), resting pressure of the lower esophageal sphincter (15.0 mm Hg and 21.5 mm Hg), and pressure of the esophageal body (78.0 mm Hg and 70.0 mm Hg). There were differences comparing the pre and post operative period in the pHmetrical findings in the following items: total number of acid refluxes (68.0 and 3.0), total number of acid refluxes more than 5 minutes (2.0 and 0.0), acid reflux more than 5 minutes in supine position (1.0 and 0.0), acid reflux more than 5 minutes in upright position (0.0 and 0.0), acidification period (5.5 and 0.1 min), DeMeester score (33.0 and 0.8). CONCLUSIONS: The esophageal manometry and pHmetry findings were improved in the post operative period with statistical significance when compared to the pre operative period. The exams were considered efficient in the evaluation of the surgical procedure. PMID- 17119666 TI - Short segment Barrett's esophagus and distal gastric intestinal metaplasia. AB - BACKGROUND: Short segment Barrett's esophagus is defined by the presence of <3 cm of columnar-appearing mucosa in the distal esophagus with intestinal metaplasia on histophatological examination. Barrett's esophagus is a risk factor to develop adenocarcinoma of the esophagus. While Barrett's esophagus develops as a result of chronic gastroesophageal reflux disease, intestinal metaplasia in the gastric cardia is a consequence of chronic Helicobacter pylori infection and is associated with distal gastric intestinal metaplasia. It can be difficult to determine whether short-segment columnar epithelium with intestinal metaplasia are lining the esophagus (a condition called short segment Barrett's esophagus) or the proximal stomach (a condition called intestinal metaplasia of the gastric cardia). AIMS: To study the association of short segment Barrett's esophagus (length <3 cm) with gastric intestinal metaplasia (antrum or body) and infection by H. pylori. PATIENTS AND METHODS: Eight-nine patients with short segment columnar-appearing mucosa in the esophagus, length <3 cm, were studied. Symptoms of gastroesophageal reflux disease were recorded. Biopsies were obtained immediately below the squamous-columnar lining, from gastric antrum and gastric corpus for investigation of intestinal metaplasia and H. pylori. RESULTS: Forty two from 89 (47.2%) patients were diagnosed with esophageal intestinal metaplasia by histopathology. The mean-age was significantly higher in the group with esophageal intestinal metaplasia. The two groups were similar in terms of gender (male: female), gastroesophageal reflux disease symptoms and H. pylori infection. Gastric intestinal metaplasia (antrum or body) was diagnosed in 21 from 42 (50.0%) patients in the group with esophageal intestinal metaplasia and 7 from 47 (14.9%) patients in the group with esophageal columnar appearing mucosa but without intestinal metaplasia. CONCLUSION: Intestinal metaplasia is a frequent finding in patients with <3 cm of columnar-appearing mucosa in the distal esophagus. In the present study, short segment intestinal metaplasia in the esophagus is associated with distal gastric intestinal metaplasia. Gastroesophageal reflux disease symptoms and H. pylori infection did not differ among the two groups studied. PMID- 17119667 TI - [Potentially pre-neoplasics areas in rat's liver associated to chronic use of phenobarbital]. AB - BACKGROUND: Phenobarbital has been used in experimental models because it is an important agent of carcinogenesis promotion in the liver of rats, and it is also non-genotoxic, organ-specific and dose-dependent. AIM: To evaluate the effects of the daily administration of phenobarbital in old rats treated with phenobarbital since their birth up to 24 months of age, in the absence of concomitant administration of chemical agents, which initiate carcinogenesis. PATIENTS AND METHODS: A control group of male Wistar rats was fed with a basic diet and a second group was fed with the same basic diet added of 0.05% of phenobarbital, for a period of 24 months. Medium and right liver fragments were submitted to the histological processing and they were stained by hematoxiciline and eosin and were immunohystochemically colored to glutathione S-transferase placentary form. RESULTS: Glutathione S-transferase placentary positive zones were detected in both groups and the images were analyzed concerning their number and surface extension through the technique of histometry analyses. CONCLUSION: Chronic use of phenobarbital did not modify the number of glutathione S-transferase placentary form positive areas. Although, data indicates that glutathione S transferase placentary form positive areas media size are increased, probably because there are an increase in their evolution capacity and irreversibility. PMID- 17119668 TI - [Study of artificial valves on left colon after rectal partial abdominoperineal resection with perineal colostomy, in dogs]. AB - BACKGROUND: Throughout time, perineal colostomy in abdominoperineal resection, as a way to avoid abdominal colostomy, was studied. Perineal colostomy associated with a slowing down mechanism for stool transit has been studied for many years with satisfactory results. AIM: The investigation of a colic valvoplasty in dogs which have undergone an abdominoperineal resection plus perineal colostomy, and the discussion of the results achieved in clinic and histopathologic analysis are the objective of this study. MATERIAL AND METHODS: The experimental model of this research studied the colon of dogs. Sixty five animals were operated and divided in five groups: 10 animals in the pilot group, 15 animals in control group and 40 animals in 3 other groups of observation with 10, 20 and 30 days of postoperative care. The dogs underwent a rectal partial abdominoperineal resection with perineal colostomy associated to a circumferential extramucosal seromiotomy of the left colon (except in the control group). RESULTS: Through postoperative and post-mortem observation it was possible to verify solid stool before the valves showing the efficiency of the mechanism in slowing it down. The histological analysis of the valves showed connective growth with fibrosis in the whole incised serous and muscular layers, separating them. A fibrous ring diminished the colon lumen in response to the artificial valve created by the scar that appeared in the inverted suture of the serous and muscular layers. The inflammatory fibrous repair process occurred in all valves, associated to foreign body type reactions and to partial stricture of the colon lumen. CONCLUSION: The circumferential seromiotomy produces a fibrous ring that provides stool retention on the descending colon. PMID- 17119669 TI - [Endoscopic treatment of squamous cell esophageal cancer]. AB - OBJECTIVE: In this article, it was evaluated the role of endoscopic procedures for the management of squamous cell esophageal cancer. DATA SOURCE: Relevant publications cited at PubMed database in the last 10 years were analyzed and compared with the experience developed at the Gastrointestinal Endoscopy Division of the Department of Gastroenterology of the University of Sao Paulo School of Medicine. Mucosectomy and advanced tumor tunnelization were the most important developments in that area. DATA SYNTHESIS: Endoscopic mucosal resection of early epidermoid cancer of the esophagus is indicated when the lesion is confined to the epithelium (m1) or to the lamina propria (m2). The described 5-year survival rate after endoscopic mucosal resection of intramucosal epidermoid tumor of the esophagus approaches 95%. Based on the available evidence, it seems reasonable to indicate endoscopic mucosal resection as a first-choice treatment for patients with intramucosal epidermoid esophageal carcinoma. There are a variety of endoscopic palliative methods for dysphagia relief in advanced esophageal cancer. CONCLUSIONS: The choice will vary according to the anatomical features and location of the tumor, patient preferences, local and expertise availability. The technical success rate for placement of metal stents across the malignant stenosis is close to 100%. The rate of long-term palliation of dysphagia approaches 80% which makes expandable metal stents the treatment of choice for palliation of obstructive symptoms caused by advanced squamous cell cancer of the esophagus. PMID- 17119670 TI - [Portuguese-language cultural adaptation and translation of "The Bowel Disease Questionnaire" used to assess functional bowel disorders]. AB - BACKGROUND: Functional bowel disorders are considered a public health problem, but there are a few prevalence studies in Brazil. In 1999, researchers from the Mayo Clinic developed The Bowel Disease Questionnaire with the purpose to evaluate functional bowel disorders. Although this has been used in different studies and population, it is often necessary to perform a cultural adaptation of a questionnaire developed for use in another culture, in order to improve the quality of the information obtained. OBJECTIVE: To translate and adapt the Bowel Disease Questionnaire to Portuguese for its use in adolescents. METHODS: Assessment of conceptual and item equivalence involved evaluation of the pertinence of the concepts in the Brazilian context; translation was evaluated through semantic equivalence between the original instrument and the Portuguese version; measurement equivalence between the original and the Portuguese version was assessed by the intra-observer reliability and internal consistency. RESULTS: The Bowel Disease Questionnaire's concepts were considered pertinent in the Brazilian context; the semantic equivalence between the original and the Portuguese version were high; reliability ranged from moderate to almost perfect; internal consistency of the scales ranged from 0,66 to 0,74. CONCLUSION: The Portuguese version of the Bowel Disease Questionnaire seems to be semantically and culturally equivalent to the original version and might be useful for measuring functional bowel disorders among Brazilian adolescents. PMID- 17119671 TI - Salmonella serovars isolated from humans in Sao Paulo State, Brazil, 1996-2003. AB - Salmonellosis remains an important cause of diarrheal illness in humans in Sao Paulo State, Brazil. In this study were identified 3554 Salmonella isolates from human infections, during the period 1996-2003. Among 68 different serovars determined, S. Enteritidis was the most frequent one in gastrointestinal and extra-intestinal infections accounting for 67.4% of all isolates. S. Typhimurium and S. enterica subsp. enterica (4,5,12:i:-) were most frequently isolated from children aged < 1-4 year-old, in contrast, people with S. Enteritidis infections were most likely to be 20-50 year-old. In our geographic area the continued laboratorial surveillance of salmonellosis, including serotyping, has showed the trends in Salmonella serovars causing infections in humans throughout the time. PMID- 17119672 TI - Virulence factors of uropathogenic Escherichia coli from a university hospital in Ribeirao Preto, Sao Paulo, Brazil. AB - The aim of the study was to determine the occurrence of virulence genes expressing fimbriae, production of hemolysin, colicin and aerobactin among a hundred Escherichia coli isolates obtained from in-and outpatients of a tertiary care teaching hospital, between July and August 2000, showing clinical and laboratory signs of urinary tract infection (UTI). The presence of genes (pap, afa, sfa) for fimbriae expression was assayed using specific primers in a polymerase chain reaction. Among the isolates studied, the prevalence of the virulence factors was 96.0%, 76.0%, 24.0%, for hemolysin, aerobactin and colicin, respectively; the prevalence of genes coding for fimbrial adhesive systems was 32.0%, 19.0% and 11.0% for pap, sfa and afa respectively. The strains isolated from the outpatients displayed a greater number of virulence factors compared to those from hospitalized subjects, emphasizing the difference between these two kinds of patients. PMID- 17119673 TI - Mass treatment for intestinal helminthiasis control in an Amazonian endemic area in Brazil. AB - The objective of the present study was to estimate the prevalence of soil transmitted helminthiasis and evaluate the sanitary conditions and the role of a mass treatment campaign for control of these infections in Santa Isabel do Rio Negro. A cross-sectional survey was carried out in 2002, to obtain data related to the sanitary conditions of the population and fecal samples for parasitological examination in 308 individuals, followed by a mass treatment with albendazole or mebendazole with coverage of 83% of the city population in 2003. A new survey was carried out in 2004, involving 214 individuals, for comparison of the prevalences of intestinal parasitosis before and after the mass treatment. The prevalences of ascariasis, trichuriasis and hookworm infection were 48%; 27% and 21% respectively in 2002. There was a significant decrease for the frequency of infections by Ascaris lumbricoides (p < 0.05; OR / 95% CI = 0.44 / 0.30 - 0.65), Trichuris trichiura (p < 0.05; OR / 95% CI = 0.37 / 0.22 - 0.62), hookworm (p < 0.05; OR / 95% CI = 0.03 / 0.01 - 0.15) and helminth poliparasitism (p < 0.05; OR / 95% CI = 0.16 / 0.08 - 0.32). It was also noticed a decrease of prevalence of infection by Entamoeba histolytica / dispar (p < 0.05; OR / 95% CI = 0.30 / 0.19 - 0.49) and non-pathogenic amoebas. It was inferred that a mass treatment can contribute to the control of soil-transmitted helminthiasis as a practicable short-dated measure. However, governmental plans for public health, education and urban infrastructure are essential for the sustained reduction of prevalences of those infections. PMID- 17119674 TI - Echovirus 4 associated to hand, foot and mouth disease. AB - Hand, foot and mouth disease (HFMD) is a contagious enteroviral infection occurring primarily in children and characterized by vesicular palmoplantar eruptions and erosive stomatitis. Echovirus 4 (EV-4) has been commonly associated with aseptic meningitis. The association of HFMD with EV-4 has not been reported previously. Two samples of a 14-month child who presented mild fever, sores in the mouth, rash with blisters on the palm of hands and soles of feet were sent to Enteric Viruses Laboratory of Adolfo Lutz Institute. Clinical samples were inoculated in three different cell lines, and those which presented cytopathic effect (CPE), were submitted to Indirect Immunofluorescence Assay (IFA) and "one step" RT-PCR. Agarose gel electrophoresis from RT-PCR product, showed a product with 437 bp, which is characteristic of Enterovirus group. Echovirus 4 was identified by IFA. Although HFMD is a viral infection associated mainly with Enterovirus 71 (HEV-71) and Coxsackievirus A16 (CV-A16), our results demonstrate a diversity of serotype related to HFMD and stress the importance of epidemiological surveillance to this disease and its complications. PMID- 17119675 TI - Detection of HIV and HCV RNA in semen from Brazilian coinfected men using multiplex PCR before and after semen washing. AB - INTRODUCTION: Prolonged survival of patients under HAART has resulted in new demands for assisted reproductive technologies. HIV serodiscordant couples wish to make use of assisted reproduction techniques in order to avoid viral transmission to the partner or to the newborn. It is therefore essential to test the effectiveness of techniques aimed at reducing HIV and HCV loads in infected semen using molecular biology tests. METHODS: After seminal analysis, semen samples from 20 coinfected patients were submitted to cell fractioning and isolation of motile spermatozoa by density gradient centrifugation and swim-up. HIV and HCV RNA detection tests were performed with RNA obtained from sperm, seminal plasma and total semen. RESULTS: In pre-washing semen, HIV RNA was detected in 100% of total semen samples, whereas HCV RNA was concomitantly amplified in only one specimen. Neither HIV nor HCV were detected either in the swim-up or in the post-washing semen fractions. CONCLUSIONS: Reduction of HIV and/or HCV shedding in semen by density gradient centrifugation followed by swim up is an efficient method. These findings lead us to believe that, although semen is rarely found to contain HCV, semen processing is highly beneficial for HIV/HCV coinfected individuals. PMID- 17119676 TI - Clinical and epidemiological aspects of HTLV-II infection in Sao Paulo, Brazil: presence of tropical spastic paraparesis/HTLV-associated myelopathy (TSP/HAM) simile diagnosis in HIV-1-co-infected subjects. AB - In this study, the epidemiological and clinical features observed in solely HTLV II-infected individuals were compared to those in patients co-infected with HIV 1. A total of 380 subjects attended at the HTLV Out-Patient Clinic in the Institute of Infectious Diseases "Emilio Ribas" (IIER), Sao Paulo, Brazil, were evaluated every 3-6 months for the last seven years by infectious disease specialists and neurologists. Using a testing algorithm that employs the enzyme immuno assay, Western Blot and polymerase chain reaction, it was found that 201 (53%) were HTLV-I positive and 50 (13%) were infected with HTLV-II. Thirty-seven (74%) of the HTLV-II reactors were co-infected with HIV-1. Of the 13 (26%) solely HTLV-II-infected subjects, urinary tract infection was diagnosed in three (23%), one case of skin vasculitis (8%) and two cases of lumbar pain and erectile dysfunction (15%), but none myelopathy case was observed. Among 37 co-infected with HIV-1, four cases (10%) presented with tropical spastic paraparesis/HTLV associated myelopathy (TSP/HAM) simile. Two patients showed paraparesis as the initial symptom, two cases first presented with vesical and erectile disturbances, peripheral neuropathies were observed in other five patients (13%), and seven (19%) patients showed some neurological signal or symptoms, most of them with lumbar pain (five cases). The results obtained suggest that neurological manifestations may be more frequent in HTLV-II/HIV-1-infected subjects than those infected with HTLV-II only. PMID- 17119677 TI - Effect of stalk and leaf extracts from Euphorbiaceae species on Aedes aegypti (Diptera, Culicidae) larvae. AB - The objective of this study was to evaluate the larvicidal activity of essential oil aqueous solutions (hydrolates) obtained by steam distillation of stalks and leaves of Croton argyrophylloides, Croton nepetaefolius, Croton sonderianus and Croton zehntneri against Aedes aegypti larvae. Twenty-five larvae of third instar were placed in plastic beckers, containing the hydrolates (50 mL), in a four repetitions scheme. Water was used as control and the number of dead larvae was counted after 24 hours. The data obtained were submitted to Variance Analysis and Tukey test. Significant differences were observed among the hydrolates from different species and from different parts of each plant (p < 0.001). The hydrolates of stalk and leaf from C. nepetaefolius and C. zehntneri and leaf hydrolate of C. argyrophylloides presented 100% mortality against larvae. The compounds present in C. zenhtneri and C. nepetaefolius are oxygenated phenylpropanoids that are more soluble in water than the monoterpenes and sesquiterpenes detected in the oils of C. argyrophylloides and C. sonderianus. This study showed that all species analyzed presented compounds with larvicidal properties, with differences between each plant parts. PMID- 17119678 TI - Sensitivity of an immunoenzymatic test for detection of anti-L. brasiliensis antibodies compared to other tests used for the diagnosis of American cutaneous leishmaniasis. AB - The diagnosis of American cutaneous leishmaniasis (ACL) is frequently based on clinical and epidemiological data associated with the results of laboratory tests. Some laboratory methods are currently being applied for the diagnosis of ACL, among them the indirect immunofluorescence reaction (IIFR), the Montenegro skin test (MST), histopathological examination, and the polymerase chain reaction (PCR). The performance of these methods varies in a considerable proportion of patients. After the standardization of an immunoenzymatic test (ELISA) for the detection of IgG in the serum of patients with ACL using a crude Leishmania braziliensis antigen, the results obtained were compared to those of other tests routinely used for the diagnosis. The tests revealed the following sensitivity, when analyzed separately: 85% for ELISA IgG, 81% for PCR, 64.4% for MST, 58.1% for IIFR, and 34% for the presence of parasites in the biopsy. ELISA was positive in 75% of patients with ACL presenting a negative MST, in 84.8% of ACL patients with negative skin or mucous biopsies for the presence of the parasite, and in 100% of cases with a negative PCR. Thus, ELISA presented a higher sensitivity than the other tests and was useful as a complementary method for the diagnosis of ACL. PMID- 17119679 TI - Presence of Leishmania amastigotes in peritoneal fluid of a dog with leishmaniasis from Alagoas, Northeast Brazil. AB - The goal of this short communication is to report the uncommon presence of intracellular amastigotes of Leishmania in peritoneal fluid of a dog with leishmaniasis from Alagoas State, Brazil. Physical examination of an adult male rottweiler suspected to be suffering of leishmaniasis revealed severe loss of weight, ascitis, splenomegaly, moderately enlarged lymph nodes, onychogryphosis, generalized alopecia, skin ulcers on the posterior limbs, and conjunctivitis. Samples of bone marrow, popliteal lymph node, skin ulcer, and peritoneal fluid were collected and smears of each sample were prepared and stained with hematoxylin and eosin. Numerous amastigotes were detected in bone marrow, popliteal lymph node, and skin ulcer smears. Smears of peritoneal fluid revealed the unusual presence of several free and intracellular amastigotes of Leishmania. Future studies are needed to determine whether the cytology of ascitic fluid represents a useful tool for diagnosis Leishmania infection in ascitic dogs, particularly in those living in areas where canine leishmaniasis is enzootic. PMID- 17119681 TI - Does Cleome droserifolia have anti-schistosomiasis mansoni activity? AB - The present study was undertaken to assess the effect of the crude extract of Cleome droserifolia (CD) leaves on experimentally infected mice with Schistosoma mansoni. Two groups of mice, showing a patent infection of S. mansoni, one of them was daily treated with an alcoholic extract of CD leaves (0.31 g kg-1 body weight, i.p.) for 21 days. The schistosomicidal activity of the CD extract was evaluated, three weeks post-treatment, on some parasitological and histopathological aspects including worm load, oogram pattern, faecal eggs releasing and granuloma formation. In addition, serum thyroid hormones levels (tri-iodothyronine; T3 and tetra-iodo-thyronine; T4), serum total protein contents and hepatic reduced glutathione (GSH) were evaluated. Treatment using CD extract resulted in a weak reduction in worm burden (32.46%) and affected the viability of both mature and immature eggs as indicated by the increase in the percentage of dead eggs and the decrease in the percentage of live ones. In addition, a week post-treatment, eggs elimination was observed in the stool of the infected-treated group which was low compared to the infected group. There was a suppressive effect of the extract on granuloma formation that could be due to the antioxidant effect of the extract. These data are confirmed by increasing hepatic GSH, serum total proteins and thyroid hormone levels in the infected treated group as compared to the infected group. Treatment significantly enhanced b globulin fractions of the protein. Based on these assumptions, CD extract has beneficial effects on thyroid hormones status and anti-schistosomiasis activity. The beneficial effects of CD extract could be related to its direct effects on the parasite, and secondary to its effect on the antioxidant capacity of the host. The present study could emphasize the precise mechanism (s) of CD extract protection. PMID- 17119682 TI - Primary liver AIDS-related lympoma. AB - Non-Hodgkin's lymphomas (NHL) are the second most frequent malignancies in AIDS patients. The majority of NHL associated with AIDS involves extranodal sites, especially the digestive tract and the central nervous system. Primary liver lymphoma (PLL) is an uncommon neoplasm among these patients. Ultrasonography and computed tomography scans may be helpful in the diagnosis of focal hepatic lymphoma. Image-guided fine-needle biopsy with histopathology of the liver lesions is the gold standard for the diagnosis of hepatic lymphoma. We report a case of PLL as the initial manifestation of AIDS in a patient without any previous infection by hepatitis C or B virus, presented as multiple and large hepatic masses. PMID- 17119683 TI - Brain abscess secondary to frontal osteomyelitis. AB - Frontal osteomyelitis is a rare complication of sinusitis. Common intracranial complications of the frontal osteomyelitis are meningitis, epidural empyema, subdural empyema and brain abscess. We described a case of frontal osteomyelitis with brain abscess caused by Staphylococcus aureus with improve after needle aspiration and antibiotics to brain abscess for eight weeks and for chronic osteomyelitis for four months. PMID- 17119684 TI - Unpredicted HTLV-1 infection in female sex worker from Imbituba, Santa Catarina, Brazil. PMID- 17119685 TI - Does acupuncture work? Is there any evidence? PMID- 17119686 TI - Protein expression of c-erbB-2 and p53 in normal ducts, ductal carcinoma in situ and invasive carcinoma of the same breast. AB - CONTEXT AND OBJECTIVE: Breast cancer is thought to derive from progressively aberrant, non-invasive breast lesions, but it is not known exactly how invasive breast cancer develops from these lesions. The aim of this study was to verify the changes in c-erbB-2 and p53 protein expression between non-neoplastic ducts, ductal carcinoma in situ and invasive ductal carcinoma found in the same breast. DESIGN AND SETTING: This was a cross-sectional study at Centro de Atencao Integral a Saude da Mulher, Campinas, Brazil. METHODS: Fifty-six women with invasive ductal carcinoma and ductal carcinoma in situ in the same breast were included. The expression of c-erbB-2 and p53 proteins was assessed in non neoplastic and neoplastic cells using immunohistochemical techniques. RESULTS: The c-erbB-2 protein was absent in non-neoplastic ducts but was present in 46% and 36% of in situ and invasive carcinoma components, respectively. Only 2% of non-neoplastic ducts, and 18% and 16% of ductal carcinoma in situ and invasive carcinoma components, respectively, were positive for p53 protein. No significant difference in c-erbB-2 and p53 protein expression was observed between in situ and invasive components. The nuclear grade agreement between in situ and invasive carcinoma was very good. CONCLUSIONS: The invasiveness of ductal carcinoma in situ seems to be independent of the Her-2/neu and TP53 genes. The general features of an occurrence of breast carcinoma are formulated at the outset of carcinogenesis, and the Her-2/neu and TP53 genes are involved. PMID- 17119687 TI - Cigarette smokers views on their habit and the causes of their illness following lung cancer diagnosis: a clinical-qualitative study. AB - CONTEXT AND OBJECTIVE: Lung cancer is the commonest malignant tumor and is increasing in incidence by 2% a year. In 90% of diagnosed cases, it is associated with tobacco product consumption. It is the greatest cause of mortality among cancer types in Brazil. Knowledge of patients psychological representations is needed for evaluating treatments and educating patients. The aim here was to interpret how smokers with lung cancer interpret the possible causes of their illness and to understand their perceptions regarding cigarette use. DESIGN AND SETTING: Clinical-qualitative study (exploratory, non-experimental) at the Pulmonary Disease Service, General Hospital, Universidade Estadual de Campinas. METHODS: An intentional small sample of cancer inpatients was recruited. The group was closed with 11 subjects, following attainment of data saturation from interviews. These interviews were semi-directed, with in-depth open-ended questions on interviewees observations, applied in a confidential setting using a tape recorder. Interviewees responses were categorized using qualitative content analysis and the results were assessed using interdisciplinary theoretical concepts, particularly from medical psychology. RESULTS: Six males and five females aged between 46 and 68 years who presented diverse clinical conditions were interviewed. CONCLUSIONS: A broader approach towards the psychological comprehension of such patients is needed, considering that cigarette consumption involves conscious and unconscious motivations, sociocultural and educational factors, the glamour of tobacco advertising, and problems with psychophysical dependence. Such an approach would avoid the perception among patients that the healthcare team are "inquisitors". This would lead to better adherence to treatment and better quality of life. PMID- 17119688 TI - Modified radical mastectomy sparing one or both pectoral muscles in the treatment of breast cancer: intra and postoperative complications. AB - CONTEXT AND OBJECTIVE: Modified radical mastectomy is widely utilized in breast cancer treatment. However, no prospective comparison has yet been made between the Madden technique (preservation of the pectoralis minor muscle) and the Patey technique (resection of this muscle). The aim of this work was to compare these two modified radical mastectomy techniques, by analyzing their degrees of difficulty and complications. DESIGN AND SETTING: Randomized trial at the Breast Unit of Hospital Araujo Jorge, Goias; and Faculdade de Medicina da Universidade Federal de Goias. METHODS: 430 patients with breast cancer with an indication for modified radical mastectomy were included in the program, of whom 426 patients were available for analysis (225 allocated to Patey and 201 to Madden). The chi squared and Student t tests were used for analysis. RESULTS: The patients demographics were well balanced between the two groups. The mean duration of the surgical procedures was 105 (+/- 29.9) and 102 minutes (+/- 33), for the Patey and Madden groups, respectively (p = 0.6). Hospitalization duration was 2.3 days for both groups. The mean number of lymph nodes resected was 20.3 (+/- 7.6) for Patey and 19.8 (+/- 8.1) for Madden (p = 0.5). There were no differences in terms of vascular or nerve sections, hematomas or infections. The surgeons reported the same degree of difficulty for the two methods. CONCLUSION: The removal of the pectoralis minor muscle did not influence any of the variables studied. Therefore, either technique can be performed, at the surgeon's discretion. PMID- 17119689 TI - Antipyretic efficacy and tolerability of oral ibuprofen, oral dipyrone and intramuscular dipyrone in children: a randomized controlled trial. AB - CONTEXT AND OBJECTIVE: Dipyrone is a widely used over-the-counter antipyretic in Latin America, and elsewhere among Latin immigrants. Despite limited evidence, physicians often prescribe oral ibuprofen or intramuscular dipyrone as the most effective antipyretics. Our aim was to compare the antipyretic efficacy and tolerability of a single dose of oral ibuprofen, oral dipyrone or intramuscular dipyrone in febrile children. DESIGN AND SETTING: Randomized, single-blind clinical trial, at San Bartolome Mother-Child National Teaching Hospital, Lima, Peru. METHODS: Children from six months to six years old with fever (rectal temperature: 38.3 to 39.8 degrees C) in the emergency ward between February and June 2003 were eligible. Seventy-five children were randomly assigned to receive a single dose of oral ibuprofen (10 mg/kg), oral dipyrone (15 mg/kg) or intramuscular dipyrone (15 mg/kg). The primary outcome was mean temperature reduction after 30, 45, 60, 90 and 120 minutes. Secondary outcomes were fever associated symptoms and clinical adverse events. RESULTS: Fever decreased by about 0.5 degrees C after 45 minutes and by about 1.0 degrees C after 120 minutes in all three groups. Mean temperatures were similar for the three groups at all times. There was a significant decrease in fever-associated symptoms for all groups. Six patients (four receiving oral dipyrone and two receiving ibuprofen) were withdrawn because of vomiting within 20 minutes after first dose of study medication. One patient assigned to oral ibuprofen presented transient urticaria. CONCLUSIONS: Antipyretic efficacy and tolerability were similar for oral ibuprofen, oral dipyrone and intramuscular dipyrone. Oral antipyretics seem more appropriate for feverish children. PMID- 17119690 TI - Diagnosis of spontaneous bacterial peritonitis in cirrhotic patients in northeastern Brazil by use of rapid urine-screening test. AB - CONTEXT AND OBJECTIVE: Spontaneous bacterial peritonitis (SBP) is a frequent and severe complication of cirrhotic patients with ascites. It has been proposed that the reagent strip for leukocyte esterase designed for the testing of urine (Combur test UX) could be a useful tool for diagnosing SPB. The aim of this study was to assess the sensitivity and specificity of urine test strips for diagnosing SBP in cirrhotic patients with ascites. DESIGN AND SETTING: Prospective study, at a university hospital in northeastern Brazil. METHODS: Forty-two unselected consecutive cirrhotic patients (32 males; mean age: 51.7 +/- years) were included, and a total of 100 paracenteses were performed. All ascitic fluid samples were analyzed using the reagent strip and cytology, neutrophils, lymphocyte count, appropriate biochemical tests and culturing. The strips were considered positive if the color became purple on a colorimetric scale. RESULTS: Nine patients were diagnosed with SBP using cytology (> 250 neutrophils/mm(3)), and the strips were positive for all these nine patients with SBP. In one sample, the strip was positive but the neutrophil count was less than 250 cells/mm(3). For 86 samples, both the strips and cytology were negative. At the threshold of 250 neutrophils/mm(3) in ascitic fluid, the sensitivity, specificity, positive predictive value and negative predictive value for the strips were respectively 100%, 98.9%, 92.3% and 100%. CONCLUSION: The Combur test UX urine screening test is a very sensitive and specific method for diagnosing SBP in cirrhotic patients with ascites. PMID- 17119691 TI - Low sensitivity of three-phase bone scintigraphy for the diagnosis of repetitive strain injury. AB - CONTEXT AND OBJECTIVE: The diagnosis of repetitive strain injury (RSI) is subjective and solely based on clinical signs and physical examination. The aim of this paper was to assess the usefulness of three-phase bone scintigraphy (TPBS) in diagnosing RSI. DESIGN AND SETTING: Prospective study at the Division of Nuclear Medicine, Department of Radiology, School of Medical Sciences, Universidade Estadual de Campinas (UNICAMP). METHODS: Seventy-three patients (mean age 31.2 years; 47 males) with clinical suspicion of RSI in the upper limbs were studied. A total of 127 joints with suspicion of RSI were studied. The shoulders, elbows and wrists were analyzed semi-quantitatively, using the shafts of the humeri and ulnae as references. The results were compared with a control group of 40 normal individuals. The patients signs and symptoms were used as the "gold standard" for calculating the probabilities. RESULTS: From visual analysis, abnormalities were observed in the flow phase for four joints, in the blood pool phase for 11 joints and in the delayed images for 26 joints. Visual analysis of the joints of the control group did not show any abnormalities. Semi-quantitative analysis showed that most of the patients joint ratios were normal. The exceptions were the wrists of patients with left-sided RSI (p = 0.0216). However, the sensitivity (9%) and accuracy (41%) were very low. CONCLUSION: TPBS with semi quantitative analysis has very low sensitivity and accuracy in the detection of RSI abnormalities in the upper limbs. PMID- 17119692 TI - Experience with local negative pressure (vacuum method) in the treatment of complex wounds. AB - CONTEXT AND OBJECTIVE: Use of topical negative pressure over difficult-to-heal wounds has been studied. The objective of this study was to analyze the effects from negative pressure in the treatment of complex wounds. DESIGN AND SETTING: Case series developed at Hospital das Clinicas, Faculdade de Medicina da Universidade de Sao Paulo. METHODS: Twenty-nine patients with complex wounds were treated with the vacuum system and then underwent a surgical procedure to close the wound. RESULTS: 85% of the skin grafts took well, and 87.5% of the local flaps were successful, thus demonstrating adequate wound preparation. The wounds were closed within shorter times than observed using other conventional treatments In two cases, the vacuum system was also used to stabilize the skin grafts over the wounds. CONCLUSIONS: Use of the vacuum method is safe and efficient for preparing wounds for surgical closure. It allows for an improvement of local wound conditions, and healthy granulation tissue develops with control over local infection. PMID- 17119694 TI - Laryngeal papillary carcinoma with unexpected evolution: case report. AB - CONTEXT: According to the literature, laryngeal papillary carcinoma is rare and has a benign prognosis. CASE REPORT: In this report we present a surprising case with nodal metastasis at the time of diagnosis. Computed tomography showed infiltration of the lesion and metastatic lymph nodes. The resected specimen was submitted to histopathological study that confirmed the diagnosis of papillary squamous cell carcinoma. PMID- 17119693 TI - Detection of free circulating Epstein-Barr virus DNA in plasma of patients with Hodgkin's disease. AB - CONTEXT AND OBJECTIVE: Free circulating Epstein-Barr virus (EBV) DNA is often present in the plasma of Hodgkins disease patients. The aim here was to evaluate the prevalence of this finding, its correlation with the immunohistochemical expression of LMP-1 (latent membrane protein 1) and the influence of other clinical factors. DESIGN AND SETTING: Prospective study in two public tertiary institutions: Hematology Service, Universidade Federal do Rio de Janeiro, and Oncology Service, Instituto Nacional do Cancer, Rio de Janeiro. METHODS: A cohort of 30 patients with newly diagnosed Hodgkins disease was studied. The control group consisted of 13 healthy adult volunteers. EBV DNA was determined by conventional polymerase chain reaction (PCR). RESULTS: The median age was 28 years, and 16 patients were women. Advanced disease was present in 19 patients, and six were HIV-positive. EBV DNA was present in the plasma of 13 patients and one control (43% versus 8%, p = 0.03). EBV DNA prevalence was higher in HIV positive patients (100% versus 29%, p = 0.0007) and those with advanced disease (63% versus 9%, p = 0.006). Among HIV-negative patients alone, EBV DNA prevalence remained higher in those with advanced disease. EBV DNA was found in 10/11 patients with LMP-1 expression in the lymph nodes, and in 3/19 without LMP-1 expression (kappa coefficient = 0.72). CONCLUSION: EBV DNA was present in 91% of patients with EBV-associated Hodgkins disease, and in all patients with HIV associated Hodgkins disease. EBV DNA prevalence was higher in patients with advanced disease, irrespective of HIV status. PMID- 17119695 TI - Pure sarcomatous recurrence of clear cell renal carcinoma following radical nephrectomy and dendritic cell vaccination. AB - CONTEXT: Sarcomatous differentiation, which represents transformation to high grade malignancy, can occur in all histological types of renal malignancy. CASE REPORT: The authors report on the case of a 66-year-old woman with a right renal mass that was shown to be a clear cell carcinoma. She underwent radical nephrectomy and dendritic cell vaccination and, 3.5 years later, she developed retroperitoneal pure sarcomatous recurrence of the tumor. The authors speculate that the vaccination could have played some role in this differentiation or selection of the sarcomatous component of the primary tumor. PMID- 17119696 TI - Facial pain of cardiac origin: a case report. AB - CONTEXT: Cardiac pain may radiate to the face and lead patients to seek dental care. Dentists may contribute towards the diagnosing of ischemic heart disease and thus refer patients for cardiological evaluation. CASE REPORT: A 50-year-old female patient was referred to a dentist for evaluation of a suspected temporomandibular disorder after repeated visits to medical emergency departments due to excruciating facial and left temporal pain associated with exertion. The pain would start in the chest and radiate to the neck, face and left temporal region. The patient's chief complaint was the facial pain; hence, she sought dental care. The dental examination revealed an edentulous upper jaw and partially edentulous lower jaw with full upper prosthetic set of teeth and decreased vertical dimension. X-ray of facial bones did not reveal any bone abnormalities. A diagnosis of temporomandibular disorder was made. However, she was referred for cardiological evaluation, since her pain was starting in the chest and because she had a past medical history of surgical treatment for coronary artery disease. A diagnosis of angina pectoris was made, the therapeutic regimen was optimized and her angina was brought under control. PMID- 17119697 TI - Investigation of possible association between Ser9Gly polymorphism of the D3 dopaminergic receptor gene and response to typical antipsychotics in patients with schizophrenia. AB - Typical antipsychotics have a high affinity for dopamine receptors. It is therefore of interest to investigate such loci in pharmacogenetic studies on psychosis. We investigated the hypothesis that Ser9Gly polymorphism of the DRD3 gene may play a role in the differences in individual response to typical antipsychotics between schizophrenic patients. The sample was composed of 53 good responders and 59 poor ones. No significant differences between the good and poor responders were found in the allelic distribution (good responders: Ser9 61.32%, Gly9 38.67%; poor responders: Ser9 64.40%, Gly9 35.59%; odds ratio, OR = 0.88, 0.49 < OR < 1.56; chi2 = 0.23, 1 degree of freedom, df, p = 0.63) and genotype distribution (good responders: Ser9/Ser9 37.73%, Ser9/Gly9 47.16%, Gly9/Gly9 15.09%; poor responders: Ser9/Ser9 42.37%, Ser9/Gly9 44.06%, Gly9/Gly9 13.55%; chi2 = 0.25, 2 df, p = 0.88). Nor was there any association with homozygosity (good responders: homozygous: 52.82%, heterozygous: 47.16%; poor responders: homozygous: 55.92%, heterozygous: 44.06%; odds ratio, OR = 0.88, 0.39 < OR < 1.99; chi2 = 0.11, 1 df, p = 0.74). The results did not support the hypothesis that Ser9Gly polymorphism of the DRD3 gene influences the response to typical antipsychotics in our sample of schizophrenics. PMID- 17119698 TI - Parkinson's disease and dopamine transporter neuroimaging: a critical review. AB - Parkinson's disease (PD) is a common neurodegenerative disorder that is mainly caused by dopaminergic neuron loss in the substantia nigra. Several nuclear medicine radiotracers have been developed to evaluate PD diagnoses and disease evolution in vivo in PD patients. Positron emission tomography (PET) and single photon computerized emission tomography (SPECT) radiotracers for the dopamine transporter (DAT) provide good markers for the integrity of the presynaptic dopaminergic system affected in PD. Over the last decade, radiotracers suitable for imaging the DAT have been the subject of most efforts. In this review, we provide a critical discussion on the utility of DAT imaging for Parkinson's disease diagnosis (sensitivity and specificity). PMID- 17119699 TI - Helicobacter pylori in the oral mucosa of patients submitted to allogeneic haematopoietic stem cell transplantation. AB - This study was designed to investigate the impact of haematopoietic stem cell transplantation (HSCT) on Helicobacter pylori colonization of the oral mucosa by nested polymerase chain reaction (nested-PCR). Forty six consecutive patients submitted to HSCT and 46 healthy volunteers were included in the study. Oral swabs were taken from the oral mucosa of the patients and control group. The medical records of the patients were reviewed and the following information was retrieved: gender and age of the patient, donor gender, primary disease, stem cell source (bone marrow or blood stem cells), leukocyte, neutrophil and platelet counts, and chronic graft versus host disease (cGVHD) of salivary glands. The results demonstrated an increased frequency of H. pylori in the oral mucosa of HSCT patients compared to controls (rho = 0.002). The presence of H. pylori in the oral mucosa was not related to the severity of cGVHD. The median counts of platelet/mm3, leukocytes/mm3 and neutrophils/mm3 in the group of HSCT patients positive for H. pylori were not statistically different from those of the patients negative for it. In conclusion, the present study shows increased frequency of H. pylori in the oral mucosa of HSCT patients compared to non transplanted healthy volunteers. PMID- 17119700 TI - Comparative therapeutic use of Risedronate and Calcarea phosphorica--allopathy versus homeopathy--in bone repair in castrated rats. AB - Osteoporosis, a disease characterized by progressive bone loss, has been the target of several studies in the past few years. It results in a much higher risk for fractures and might cause slower bone lesion healing. The aim of this work was to study the effects of Risedronate (allopathic medicine) and Calcarea phosphorica 6CH (homeopathic medicine) on the repair of bone lesions in male rats with osteoporosis induced by castration. Eighty-four three-month-old rats were used divided into four groups of twenty-one animals each. Three groups where castrated and one group was submitted to Sham surgery. One month later, cortical lesions were made in all animals' tibiae and, after one day, the different experimental treatments began according to the following groups: CR- castrated/Risedronate (1 mg/kg/day); CCp--castrated/Calcarea phosphorica 6CH (3 drops/day); CP--castrated/placebo and SP--Sham/placebo. The animals were sacrificed at seven, fourteen and twenty-eight days after the beginning of the treatments and had their tibiae removed. Digital radiographs of the tibiae were taken and analyzed in order to evaluate the optical density of the defect area. Then, they were decalcified and processed for histological and histomorphometrical analysis. The data were submitted to ANOVA, and to the Tukey and Dunnett tests (5%). The allopathic and homeopathic treatments led to different bone formation as regards remodeling and maturation aspects. Further research is necessary to access the resistance and quality of the newly formed bone. PMID- 17119701 TI - In vitro minocycline activity on superinfecting microorganisms isolated from chronic periodontitis patients. AB - Chronic periodontitis is the most common type of periodontitis and it is associated with various species of microorganisms. Enteric rods, Pseudomonas, Staphyloccocus and Candida have been retrieved from periodontal pockets of patients with chronic periodontitis and correlated to cases of superinfection. Local or systemic antibiotic therapy is indicated to reinforce the effects of the conventional mechanical therapy. Minocycline has been suggested as one of the most effective drugs against periodontal pathogens. The aim of this work was to evaluate the minimal inhibitory concentration (MIC) of minocycline on superinfecting microorganisms isolated from the periodontal pocket and the oral cavity of individuals with chronic periodontitis. Isolates of Enterobacteriaceae (n = 25), Staphylococcus spp. (n = 25), Pseudomonas aeruginosa (n = 9) and Candida spp. (n = 25) were included in the study. Minimal inhibitory concentrations (MIC) of minocycline were determined using the Mueller-Hinton agar dilution method. Staphylococcus spp. isolates were the most sensitive to minocycline with a MIC of 8 microg/mL, followed by Enterobacteriaceae with a MIC of 16 microg/mL. The concentration of 16 microg/mL inhibited 96% of Candida spp. isolates. The MIC for 88.8% of the isolates of Pseudomonas aeruginosa was 128 microg/mL. A concentration of 1,000 microg/mL was not enough to inhibit 100% of the tested isolates. PMID- 17119702 TI - Reproducibility of pixel values for two photostimulable phosphor plates in consecutive standardized scannings. AB - The objective of the present study was to determine the reproducibility of the pixel values obtained with the Digora system (Soredex, Finland). Exposures were standardized, with variation in exposure and scanning time of two photostimulable phosphor plates containing a stepwedge image. The smallest variation in pixel values ranged from 50 to 75%, with the widest variations being observed in less dense steps. A significant difference in pixel values was observed in terms of X ray exposure and scanning times and between the two plates themselves (ANOVA, p < 0.01). Using the present methodology, the reproducibility of pixel values was not satisfactory for the tested white photostimulable plates. This wide variation in digitalization might be influenced by the amount of X-rays that sensitized the plates. It may be important to establish the reproducibility of the pixel values in quantitative studies using digital image. PMID- 17119703 TI - Diametral tensile strength of composite resins submitted to different activation techniques. AB - The aim of this study was to evaluate the diametral tensile strength (DTS) of composite resins submitted to different curing techniques. Four composite resins were tested in this study: Targis (Ivoclar), Solidex (Shofu), Charisma (Heraeus Kulzer) and Filtek Z250 (3M Espe). Sixty-four cylindrical specimens were prepared and divided into eight groups according to each polymerization technique (n = 8). The indirect composite resins (Targis and Solidex) were polymerized with their respective curing systems (Targis Power and EDG-lux); Charisma and Filtek Z250 were light-cured with conventional polymerization (halogen light) and additionally, with post-curing systems. Specimens were stored in artificial saliva at 37 degrees C for one week. DTS tests were performed in a Universal Testing Machine (0.5 mm/min). The data were statistically analyzed by ANOVA and Duncan tests. The results were (MPa): Z250/EDG-lux: 69.04 feminine; Z250/Targis Power: 68.57 feminine; Z250/conventional polymerization: 60.75b; Charisma/Targis Power: 52.34c; Charisma/conventional polymerization: 49.17c; Charisma/EDG-lux: 47.98c; Solidex: 36.62d; Targis: 32.86d. The results reveal that the post-cured Z250 composite resin showed the highest DTS means. Charisma composite presented no significant differences when activation techniques were compared. Direct composite resins presented higher DTS values than indirect resins. PMID- 17119704 TI - Biological response of pulps submitted to different capping materials. AB - Pulp capping is a procedure that comprises adequate protection of the pulp tissue exposed to the oral environment, aiming at the preservation of its vitality and functions. This study evaluated the response of the dental pulps of dog teeth to capping with mineral trioxide aggregate (MTA) or calcium hydroxide P.A. For that purpose, 37 teeth were divided into two groups, according to the capping material employed. Two dogs were anesthetized and, after placement of a rubber dam, their pulps were exposed in a standardized manner and protected with the experimental capping materials. The cavities were then sealed with resin-modified glass ionomer cement and restored with composite resin. After sixty days, the animals were killed and the specimens were processed in order to be analyzed with optic microscopy. It was observed that MTA presented a higher success rate compared to calcium hydroxide, presenting a lower occurrence of infection and pulp necrosis. PMID- 17119705 TI - Histologic evaluation of pulpotomy performed with ethyl-cyanoacrylate and calcium hydroxide. AB - The aim of this study was to evaluate histological aspects of the pulp-dentin complex of dogs submitted to pulpotomy and capped with ethyl-cyanoacrylate and calcium hydroxide. Thirty dog teeth were divided into 2 groups of 15 as follows: Group 1 - ethyl-cyanoacrylate; Group 2 - calcium hydroxide. The pulpotomies were carried out following all of the treatment precautions recommended for dogs. After 30 days the specimens were submitted to histological preparation and were then blindly evaluated by a histologist. Data were analyzed statistically by the Fisher exact test, comparing the two groups. After 30 days, the presence of a hard tissue barrier was observed in 83.3% of Group 1, and in 100% of Group 2 (p = 0.478). A continuous hard tissue barrier was observed in 50% of the ethyl cyanoacrylate group and 75% of the calcium hydroxide group (p = 0.652). It can be concluded that both materials induced hard tissue barrier formation, but Group 2 had a higher percentage than Group 1, with no significant statistical differences; the differences observed between the different barriers (continuous/non-continuous) were not significant between groups and there was no pulpal necrosis in either group. PMID- 17119706 TI - A scanning electron microscopic evaluation of different root canal irrigation regimens. AB - The purpose of this study was to assess the effectiveness of endodontic irrigants in removing the smear layer from instrumented root canal walls using Scanning Electron Microscopy (SEM). The endodontic irrigants used were: 1% sodium hypochlorite (NaOCl); 1% NaOCl mixed to 17% EDTAC; 2% chlorhexidine gel; and Ricinus communis gel. Photomicrographs of the middle and apical thirds were evaluated with the aid of the Fotoscore - v. 2.0 software. The results indicated that the mixture of sodium hypochlorite and EDTAC completely removed the smear layer from dentinal walls. The other endodontic irrigants were not as efficient in cleansing the root canals. PMID- 17119707 TI - Dimensional stability of distances between posterior teeth in maxillary complete dentures. AB - The aim of this study was to assess the displacement of posterior teeth in maxillary complete dentures stored in water at 37 degrees C. Twenty acrylic resin based maxillary complete dentures were constructed with the anterior teeth arranged in normal overlap and the posterior teeth in Angle class I. Metallic pins were placed on the labial cusp of the first premolars (PM), and on the mesiolabial cusp of the second molars (M). The final acrylic resin pressing was made in a metallic flask with aid of the RS tension system, and polymerized in a moist-hot cycle at 74 degrees C for 9 hours. The dentures were deflasked after cooling in their own polymerizing water or after cooling in polymerizing water plus bench storage for 3 hours, and stored in water at 37 degrees C for periods of 7, 30, and 90 days. Following deflasking and after each storage period tested, the PM-PM (premolar to premolar), M-M (molar to molar), LPM-LM (left premolar to left molar), and RPM-RM (right premolar to right molar) distances were measured with an STM Olympus microscope, with an accuracy of 0.0005 mm. Collected data were submitted to ANOVA and Tukey's test (5%). There was no statistically significant difference for the PM-PM, M-M, and LPM-LM distances after all storage periods when the flask cooling methods were considered. With exception of the RPM RM distance after the 30-days water plus bench storage period, the other distances remained statistically stable. PMID- 17119708 TI - The pediatric patient at a maxillofacial service: Eye prosthesis. AB - Congenital absence or loss of the ocular globe during childhood causes psycho social and cosmetic disorders and compromise the normal development of the orbital region. The literature relating to congenital or acquired etiology, due to trauma or disease, demonstrates the necessity of prevention and early detection in order to minimize the sequelae and disturbances in orbital growth. Installation of an eye prosthesis is essential to the rehabilitation process, so as to produce satisfactory development of the region. In order to characterize a profile of the child patient with this condition, a survey was carried out at the Prosthetic Eye sector, Out-patient Clinic, Discipline of Maxillofacial Prosthodontics, School of Dentistry, University of Sao Paulo (FOUSP), during the period from 1988 to 2003. The 124 (14.02%) patients within the age group of 0-13 years registered for ocular prosthesis were divided into a first group of 64 patients (51.62%) with 0-7 years, and a second group of 60 patients (48.38%) with 8-13 years. Fifty nine were girls and 65 were boys. No statistical significance was observed regarding the distribution of genders in the two analyzed age groups (p = 0.069). However, there was statistical significance at the level of 0.01 in relation to etiology, with higher prevalence of congenital and pathological disturbances in the younger group and traumatic occurrences in the older group. The etiology also presented variation according to the gender, at the significance level of 0.05, where girls presented three times less trauma than boys in the older age group. The necessity of prosthetic ocular repair was evenly distributed along the childhood period and the eye losses that required prosthetic treatment equally affected both genders. However, the etiology of eye loss varied according to the considered gender and age bracket. PMID- 17119709 TI - Efficacy of Gluma Desensitizer on dentin hypersensitivity in periodontally treated patients. AB - The aim of this double-blind, controlled, split-mouth designed clinical trial was to assess the effect of a single application of Gluma Desensitizer on alleviating dentin hypersensitivity. Twelve subjects entered the study and ten completed the protocol. Each subject had two teeth treated: one with Gluma Desensitizer according to the manufacturer's instructions and one with water. The assessment of pain was performed with the VAS (Visual Analogue Scale), after tactile (probe), thermal (cold blast of water) and thermal/evaporative (cold blast of air) stimuli at baseline, immediately after treatment, after 1 week and after 4 weeks. The mean VAS values for the test and control teeth were compared by the paired t test (alpha = 0.05). Repeated measurements ANOVA was used to compare the different experimental times. The results showed that for test teeth, at baseline, mean VAS values were 1.76 (+/- 2.82), 7.10 (+/- 2.10) and 4.75 (+/- 2.65), and, after 4 weeks, the mean values were 1.70 (+/- 2.31), 5.50 (+/- 3.30) and 4.61 (+/- 3.14), respectively for probe, water and air stimuli. For the control teeth, at baseline, the mean VAS values were 1.86 (+/- 2.92), 6.61 (+/- 2.31) and 4.08 (+/- 2.91) and, after 4 weeks, 2.66 (+/- 3.07), 6.32 (+/- 2.94) e 4.76 (+/- 3.26). There were no statistically significant differences between test and control teeth at any time. No intra-group differences were demonstrated either. It was concluded that Gluma Desensitizer had no effect on hypersensitive teeth from periodontally treated patients for a period up to 4 weeks. PMID- 17119710 TI - Factors related to periodontal disease in a rural population. AB - To estimate the prevalence and related aspects of periodontitis in a rural area of the State of Bahia, Brazil, this cross-sectional study was carried out in the village of Matinha dos Pretos, Feira de Santana County, Bahia, among 172 subjects ranging from 20 to 60 years of age. During household visits, a full-mouth periodontal exam was performed on each subject, who also answered a questionnaire about socio-demographic, economic and health-related issues. The factors assessed were plaque index, bleeding on probing index, probing depth, gingival recession or hyperplasia measurements. Clinical attachment loss was also calculated. The multivariate logistic regression method was used to evaluate the relative contribution of these factors to the periodontitis condition. The prevalence of periodontitis was 24.4%. The following factors were all positively associated with the presence of periodontitis: being male (OR = 1.58; 1.00 - 2.53), being 30 years of age or older (OR = 2.80; 1.00 - 7.39), living in a house where there was more than one person per room (OR = 1.53; 0.96 - 2.45), being a cigarette or pipe smoker or ex-smoker (OR = 1.49; 0.92 - 2.39), having a plaque index of over 65% (OR = 2.97; 2.72 - 7.39) and more than four missing teeth (OR = 1.51; 0.82 - 2.78). The authors concluded that socioeconomic and biological factors, especially poor oral hygiene and older age, are positively associated with periodontitis in the rural population of a small village in the county of Feira de Santana, State of Bahia, Brazil. PMID- 17119711 TI - Translation into Brazilian Portuguese, cultural adaptation and validation of the oral health impact profile (OHIP-49). AB - The aim of this study is to develop a Portuguese version of the OHIP-49 to be used in Brazil. The adopted methodology involves translation, back-translation, revision of the first version, cultural adaptation and evaluation of its measurement properties. The questionnaire was administered in interviews with 60 year-old and older Brazilian people. The Kendall-tau correlation coefficient was used to assess reproducibility; the Cronbach alpha coefficient was used to verify internal consistency and the Kruskal-Wallis and Mann-Whitney U tests were used to assess validity. Reproducibility was shown to be statistically significant, with coefficients ranging from 0.72 to 0.74 between dimensions of three interviews. The Cronbach alpha coefficients were 0.96 and 0.90 respectively for the dimensions of the interviews and for total items. Constructive validity, demonstrated through a comparison between the questionnaire dimensions and the clinical parameters, showed statistical significance for "presence of carious lesion" (p < 0.05). Significant correlations between similar dimensions of the OHIP-49 and the SF-36 were also identified (p < 0.05). In the present sample, the Portuguese version of the OHIP-49 presented results similar to those found in the literature and proved to be a reproducible and valid parameter for evaluating the impact of oral conditions on the quality of life of Brazilians. PMID- 17119712 TI - Analysis of fluoride concentration in mother's milk substitutes. AB - The aim of the present study was to determine the fluoride concentration in some brands of mother's milk substitutes and evaluate the possibility of developing dental fluorosis by consuming these products. The products, all powdered, were divided into 3 groups: infant formulae (group I, n = 7), milk-based (group M, n = 8) and soy-based (group S, n = 3). Samples from 3 cans of different batches of each brand were reconstituted in deionized water and analyzed using the specific electrode method, after hexamethyldisiloxane (HMDS) facilitated diffusion. The fluoride content (mg F/L) of the products ranged from 0.044 to 0.326 (I), 0.014 to 0.045 (M) and 0.253 to 0.702 (S). There was significant difference in the fluoride content of cans from distinct batches (p < 0.05) in most of the brands. The reconstitution of all products in water with optimal fluoride concentration for consumption during the mineralization phase of the primary teeth could result in daily fluoride intake above 0.07 mg F/kg body weight/day. Therefore, the consumption of these products, especially when reconstituted with optimally fluoridated water, could increase the risk of developing dental fluorosis. PMID- 17119713 TI - [Articles in English in the Brazilian Archives of Ophthalmology: a result of globalization]. PMID- 17119714 TI - [Questionnaire of the psychosocial profile of the patient with anophthalmia with indication of ocular prosthesis]. AB - The patient with anophthalmia may present feelings of inferiority and rejection. Knowing his/her needs and expectations contribute to a better technical intervention. PURPOSE: To elaborate a questionnaire of the psychosocial profile of the patient with anophthalmia with indication of ocular prosthesis. METHODS: An exploratory research was used to elaborate the questionnaire, by means of a guided interview followed by writing down what was said by the interviewees, who were adult patients of the Bucco-Maxillo-Facial Prosthesis Center of FOSJC - UNESP. The guided interview was made up of 14 items directly related to the future outline of the profile. Each item of the interview resulted in questions of the questionnaire, which was pretested twice before reaching its final version. RESULTS: The patients reported, in the exploratory research, unpleasant feelings with the loss of the eye; relationship shyness; expectations regarding surgery and prosthesis use; a wish to receive explanations and to hold their opinion about the treatment. The questionnaire of the psychosocial profile of the patient with anophthalmia with indication of ocular prosthesis is, therefore, made of 43 questions divided into 5 blocks in order to aid the comprehension of the inquired aspects and to facilitate both the computation of data and discussion, and also to improve the selection of questions according to the objective of the researcher or professional. CONCLUSIONS: It was concluded that the questionnaire was viable, can be used in full or by selecting blocks and provide a panorama of the patient's history related to the problem he/she faces, from the loss of the ocular globe to the confection of the prosthesis. PMID- 17119715 TI - [Iridociliary cysts on ultrasound biomicroscopic examinations]. AB - PURPOSE: To present the prevalence of iridociliary cysts in eyes of patients submitted to UBM examination. METHODS: We analyzed retrospectively the UBM images of 1157 patients in the period from September 1995 to June 2004. The inclusion criterion was the UBM examination had been made on the four quadrants (superior, inferior, nasal and temporal) of the eyes. The cysts were assessed and classified in relation to their number in each quadrant; localization and morphology. Using the UBM Pro 2000 software we took the measurements of the largest diameter, wall thickness and area of the cysts besides the angle recession area (ARA). RESULTS: The prevalence of the cysts was 4.9%. No statistical difference was found between the prevalence of cysts regarding gender or between the right and left eyes. They were more prevalent in the inferior (39.8%) and temporal (29.1%) quadrants. The cysts with the largest diameter and area were located in the temporal and superior quadrants. No difference was found in relation to cysts' wall thickness in all quadrants. CONCLUSIONS: No statistical difference was found between the prevalence of cysts as concerns gender or between the right and left eyes. However, the cysts were more prevalent in the inferior and temporal quadrants. The locations of the largest cysts are preferentially the temporal and superior quadrants. The angle recession area was smaller in the temporal and superior quadrants. The cysts' wall thickness is practically the same in all quadrants. The diagnosis of iridociliary cysts was an occasional finding in 64.3% of the patients. PMID- 17119716 TI - Correlation between the ibopamine provocative test and the diurnal tension curve in glaucoma patients. AB - PURPOSE: To correlate the ibopamine provocative test with the diurnal tension curve (highest intraocular pressure-IOP and range) in glaucoma. METHODS: This is a prospective case series including glaucoma patients from the Federal University of Goias, Glaucoma Service. Two 2% ibopamine eyedrops were instilled into one or both eyes of each patient, 5 minutes apart. Intraocular pressure was checked before and 30 and 45 minutes after the second ibopamine instillation. Thereafter, the diurnal tension curve of each patient was assessed with five independent measurements (at every 2:30 hours), from 8:00 o'clock AM to 6:00 o'clock PM. Pearson's correlation coefficient was used to test the linear relation between the intraocular pressure after the ibopamine instillation with the highest intraocular pressure value and the intraocular pressure range in the diurnal curve. RESULTS: Thirty-one eyes from 22 patients were included. There was a significant correlation between the intraocular pressure 30 and 45 minutes after ibopamine instillation and the highest intraocular pressure assessed in the diurnal curve (r=0.356, p=0.04 and r=0.429, p=0.01, respectively). However, no correlation between IOP after the use of ibopamine and the diurnal intraocular pressure range at 30 (r=0.046, p=0.8) and 45 minutes (r=0.109, p=0.5) was observed. CONCLUSION: The ibopamine provocative test shows a significant correlation with the highest intraocular pressure in the diurnal tension curve in glaucoma patients. However, no correlation was observed with the intraocular pressure range. PMID- 17119717 TI - Genetics and prevention of blindness. AB - PURPOSE: To verify the perception of Brazilian ophthalmologists regarding the role played by Genetics in their routine medical activity and their conduct when dealing with patients, with hereditary diseases who need genetic counseling. METHODS: A cross-sectional survey was conducted by means of invitations to participate in an interview on this subject. The questionnaires were sent to 200 ophthalmologists who work in the area of Campinas, SP, Brazil. RESULTS: Among the 73 professionals who answered the questionnaire (36%), there was a high rate of positive answers (49-88%) concerning their knowledge of genetics and a low rate (10-33%) of use of this knowledge in their clinical practice. The frequency of genetic ophthalmopathies in clinical practice was relevant in the opinion of 68% of the interviewees; 92% indicated a relevant relationship between genetically determined ocular disorders and the prevention of blindness through detection, early treatment and genetic counseling. More extensive genetic knowledge was considered indispensable to 84% of the professionals, but 16% answered that it was irrelevant. CONCLUSIONS: Although these ophthalmologists have basic notions of genetics (88,0%) and are aware of its importance in the prevention of blindness, a great majority of Brazilian ophthalmologists have not acquired adequate knowledge of genetics and, in practice, rarely use its therapeutic and preventive potential. PMID- 17119718 TI - [Development of ophthalmologic software for handheld devices]. AB - INTRODUCTION: The formulas for calculation of intraocular lenses have evolved since the first theoretical formulas by Fyodorov. Among the second generation formulas, the SRK-I formula has a simple calculation, taking into account a calculation that only involved anteroposterior length, IOL constant and average keratometry. With the evolution of those formulas, complexicity increased making the reconfiguration of parameters in special situations impracticable. In this way the production and development of software for such a purpose, can help surgeons to recalculate those values if needed. PURPOSE: To idealize, develop and test a Brazilian software for calculation of IOL dioptric power for handheld computers. METHODS: For the development and programming of software for calculation of IOL, we used PocketC program (OrbWorks Concentrated Software, USA). We compared the results collected from a gold-standard device (Ultrascan/Alcon Labs) with the simulation of 100 fictitious patients, using the same IOL parameters. The results were grouped for ULTRASCAN data and SOFTWARE data. Using SRK/T formula the range of those parameters included a keratometry varying between 35 and 55D, axial length between 20 and 28 mm, IOL constants of 118.7, 118.3 and 115.8. RESULTS: Using Wilcoxon test, it was shown that the groups do not differ (p=0.314). We had a variation in the Ultrascan sample between 11.82 and 27.97. In the tested program sample the variation was practically similar (11.83-27.98). The average of the Ultrascan group was 20.93. The software group had a similar average. The standard deviation of the samples was also similar (4.53). CONCLUSION: The precision of IOL software for handheld devices was similar to that of the standard devices using the SRK/T formula. The software worked properly, was steady without bugs in tested models of operational system. PMID- 17119719 TI - [Endothelial damage after planned extracapsular cataract extraction and phacoemulsification of hard cataracts]. AB - PURPOSE: To evaluate and compare the endothelial damage after planned extracapsular cataract extraction (ECCE) and phacoemulsification of very hard cataracts. METHODS: In this prospective, randomized study, 41 patients with age related and very hard cataract were divided into two groups: in group 1 (21 patients) an extracapsular cataract extraction was performed and in group 2 (20 patients), phacoemulsification. In both groups, intraocular lenses were implanted in the capsular bag. Preoperatively and 1, 3 and 6 months postoperatively, a complete ophthalmological examination, endothelial specular microscopy, and ultrasonic pachymetry were done. Endothelial cell loss, pleomorphism, polymegathism and corneal thickness were studied. RESULTS: Both groups presented an endothelial cell loss in the postoperative time, as compared with the preoperative values, but there were no significant differences among the postoperative values (1, 3 and 6 months). Six months after surgery, mean cell loss was 28.50% in group 1 and 34.77% in group 2. There were no differences among the indexes of pachymetry, polymegathism and pleomorphism between the two groups. CONCLUSIONS: Endothelial response was not statistically different between the two studied groups. PMID- 17119720 TI - [Comparison between the postoperative results of triple procedure and combined penetrating keratoplasty/transsclerally sutured posterior chamber lens implantation]. AB - PURPOSE: To compare the outcomes of two surgical techniques of penetrating keratoplasty with different surgical time, regarding the crystalline and the intraocular lens. METHODS: This retrospective study included 37 patients' eyes divided into 2 groups: extracapsular cataract extraction, posterior chamber intraocular lens implantation and penetrating keratoplasty (Group 1, G1) and transscleral fixation of posterior chamber lens and penetrating keratoplasty (Group 2, G2). The following parameters were recorded: visual acuity, intraocular pressure, refractive astigmatism, complication and keratometric astigmatism. RESULTS: Visual acuity improved in the two groups (G1 p<0.001 and G2 p=0.008). In G2 a significant change for the worse of intraocular pressure outcome was observed when compared with the other group (p=0.014). Regarding refractive and keratometric astigmatism no significant differences between the groups were found. The follow-up was 11 months. CONCLUSION: The most important negative prognostic factor affecting visual acuity was the postkeratoplasty corneal curvature which can not be predicted accurately and the most important negative factor affecting intraocular pressure was the loss of integrity of the anterior and posterior segments. PMID- 17119721 TI - Use of optical aids by visually impaired students: social and cultural factors. AB - PURPOSE: To identify conceptions, social and cultural factors regarding the use of optical aids by visually impaired students and to present information to health and educational professionals. METHODS: Qualitative research using spontaneous theater (interactive theater modality based on improvisation) as research instrument. To analyze data, an adapted form of the collective subject discourse technique - procedures for organization of verbal data - was applied. Scenes, gestures, expressions, silences and behaviors were added to the original proposal. The study population included all visually impaired students from elementary public schools, aged 10 to 14 years who attended a resource room in a Sao Paulo state city. The students were examined at a university low vision service. RESULTS: Little knowledge about the impairment and difficult adaptation to use of optical aids were identified. The students' behavior showed denial of own problems, discomfort on public use of aids and lack of participation in own health decisions. CONCLUSION: Analysis through spontaneous theater session allows the professional to gather information which is not possible to acquire in the health assistance atmosphere. Needs, difficulties and barriers the users found before the prescribed treatment were identified. PMID- 17119722 TI - [Effects of analysis of posterior subtenon injection of triamcinolone: a case series]. AB - PURPOSE: To evaluate the effects of posterior subtenon injection of triamcinolone acetonide in patients with intraocular inflammation. METHODS: Case series enrolling 24 patients (26 eyes) submitted to posterior subtenon injection of triamcinolone acetonide for intraocular inflammation treatment. Maleate timolol at 0.5% was prescribed for all patients for 30 days. RESULTS: Improvement of visual acuity was observed in 81% of the patients after a single injection of triamcinolone acetonide. Increase in the intraocular pression was not observed in the treated eyes. CONCLUSION: Posterior subtenon injection of triamcinolone acetonide proved to be a safe and effective way of treating intraocular inflammations. PMID- 17119723 TI - Computed tomographic patterns of orbital cellulitis due to sinusitis. AB - PURPOSE: To describe the CT findings of orbital cellulitis due to sinusitis. METHODS: The records and CT scans of 45 consecutive patients with orbital cellulitis due to sinusitis treated at the Hospital of the Medical School of Ribeirao Preto were analyzed by a radiologist and two orbital surgeons. RESULTS: Three major types of CT changes were observed: diffuse fat infiltration, subperiosteal abscess and orbital abscess. Diffuse fat infiltration (characterized by an increased density of the extra- or intraconal fat) was seen in 11 patients (24.44%). A subperiosteal abscess was diagnosed in 28 patients (62.23%). A surgically proved orbital abscess was detected in 6 patients (13.33%). CONCLUSIONS: In all cases of orbital cellulitis due to sinusitis intraorbital changes can be detected by CT scans either as a diffuse infiltration of the orbital fat or as a detachment of the periorbita (subperiosteal abscess) or a true orbital abscess. Category I of Chandler orbital cellulitis classification (inflammatory edema) must be understood as a stage of a process that is already happening within the orbit and, as the term "preseptal cellulitis" means a palpebral infection, this designation should not be used to stage orbital cellulitis. PMID- 17119724 TI - Lack of papillomavirus (HPV) in pterygia of a Brazilian sample. AB - PURPOSE: Because the influence of papillomavirus (HPV) in pterygium pathogenesis is controversial, the aim of this study was to identify whether it is present in the pterygia lesions in our region. METHODS: Thirty-six patients with unilateral primary pterygia were submitted to excision of pterygia and a sample of normal conjunctiva. Tissues were submitted to polymerase chain reaction (PCR) evaluation for papillomavirus DNA detection. RESULTS: We were unable to detect any HPV DNA in all studied specimens. CONCLUSION: According to our results papillomarivus is not important for pterygium formation. PMID- 17119725 TI - [Botulinum toxin treatment for horizontal strabismus in children with cerebral palsy]. AB - To evaluate the efficacy of botulinum toxin injection in the treatment of horizontal strabismus in children with cerebral palsy. METHODS: A group of 24 patients, age 6 - 156 months, with cerebral palsy and horizontal strabismus (17 esotropias ranging from 25 to 45 PD and 7 exotropias ranging from 20 to 45 PD) were treated with botulinum toxin (34 medial rectus and 14 lateral rectus muscles). The children underwent full ophthalmic examination. The injection treatment was performed after sedation using Mendonca's forceps. The follow-up was done after 7 days, 15 days and monthly. Deviation less or equal of 10 PD after 6 months of follow-up was considered successful. However, if patients did not achieve this result a second application could be performed. RESULTS: In the group of esotropia (n=17) 47.1% had successful results with only one application after two years of follow-up. All of these patients had previous deviation less than or equal to 35 PD. In the exotropia group a single injection was not sufficient to achieve successful result. The side effects included: subconjutival hemorrhage in 4 (16.7%) patients, transient vertical deviation in 4 (16.7%) and transient ptosis in 22 (91.7%). CONCLUSION: The use of botulinum toxin was a good alternative in the treatment of children with cerebral palsy and esotropia. PMID- 17119726 TI - [Comparison of scanning laser polarimetry, optical coherence tomography 1 and Stratus optical coherence tomography for the detection of axonal loss in band atrophy of the optic nerve]. AB - PURPOSE: To compare the abilitiy of scanning laser polarimetry (GDx), optical coherence tomography (OCT) 1 and Stratus- optical coherence tomography to discriminate between healthy eyes and eyes with band atrophy of the optic nerve and temporal hemianopsia. METHODS: Twenty-three eyes with band atrophy of the optic nerve and 23 eyes from healthy subjects were included in this observational prospective case-control study. All eyes underwent retinal nerve fiber layer (RNFL) thickness analysis using GDx, optical coherence tomography-1 and Stratus optical coherence tomography. Mean values obtained with each equipment were compared between band atrophy and normal eyes. Receiver operating characteristic (ROC) curves and sensitivities at fixed specificities (80% e 95%) were calculated for each parameter calculated with each equipment and compared. RESULTS: When compared with healthy subjects, the parameters corresponding to the global average as well as each of the four quadrant retinal nerve fiber layer thickness in eyes with band atrophy were significantly smaller (p<0.05), than in normal eyes, with the exception of GDx's temporal thickness parameter. Comparison of the areas under ROC curves (AUCs) of the parameters from the three equipments revealed significantly greater values for the Stratus-OCT when compared to the OCT-1 in the global average and in the temporal quadrant thickness measurement. Stratus-OCT was significantly more sensitive than GDx in the global average as well as in the temporal, nasal and inferior quadrant. OCT-1 was superior to GDx only in the temporal quadrant. All three equipments revealed a similar ability to identify retinal nerve fiber layer reduction in the superior quadrant. CONCLUSIONS: The Stratus OCT showed the best ability to discriminate between eyes with band atrophy of the optic nerve and healthy eyes although all three equipments were able do identify most of the abnormal eyes. OCT-1 was inferior to Stratus-OCT and superior do GDx in the temporal quadrant. Although GDx was able identify some abnormality in most studied eyes, the equipment showed poor sensitivity in the detection of axonal loss in the nasal and temporal aspect of the optic disc and should be used with caution in neuro-ophthalmic diseases that cause retinal nerve fiber layer loss in those areas of the optic disc. PMID- 17119727 TI - Retinal thickness assessed by optical coherence tomography (OCT) in pseudophakic macular edema. AB - PURPOSE: To evaluate the usefulness of the optical coherence tomography (OCT) retinal map in angiographic pseudophakic macular edema (ACME). METHODS: This is a prospective analysis of a group of 36 pseudophakic eyes at the fifth postoperative week, submitted to optical coherence tomography 2 macular map and fluorescein angiography on the same day and, a control group of twenty-two 20/20 vision eyes with a minimum 6-month postoperative period, submitted to optical coherence tomography 2 macular map only. Exclusion criteria were diabetes, any macular pathology and unreadable optical coherence tomography or angiogram. The angiograms were divided into three groups according to the edema intensity: Group I: level 0 (absence of angiographic pseudophakic macular edema); group II: levels 1 and 2 (incomplete and complete perifoveal angiographic pseudophakic macular edema respectively) and group III: levels 3 and 4 (up to one disc diameter and greater than one disc diameter angiographic pseudophakic macular edema area respectively). The nine zones of the optical coherence tomography macular map were compared between the control and the three subgroups of the study group and, between group I and groups II and III. RESULTS: Among the 36 eyes, 23 (64%) were classified as group I, 10 (28%) as group II and three (8%) as group III. The mean thickness of the fovea zone was 185+/-15 microm for the control group, 189+/-24 microm for group I, 213+/-33 microm for group II and 455+/-38 microm for group III. Significant differences: between group III and the control or group I for all zones (p<0.01) and, between group II and control or group I for the fovea and temporal inner zones (p<0.05). CONCLUSION: The optical coherence tomography retinal map is useful in angiographic pseudophakic macular edema diagnosis. PMID- 17119728 TI - [Quality of life assessment in patients with blepharoptosis]. AB - PURPOSE: To assess the health-related quality of life in blepharoptosis patients and the impact after surgical management on their quality of life. METHODS: Twenty-one patients with congenital or acquired blepharoptosis completed a questionnaire which was adapted from the Visual Function Questionnaire (VFQ), before and after surgical management. RESULTS: Mean preoperative score was 74 and mean postoperative score was 81 and there was a statistically significant difference between mean pre- and postoperative scores of 7 (p=0.004). There was a statistically significant difference only for the mental health subdomain (p=0.001) in the evaluation of the questions (alpha=0.002) and subdomains (alpha=0.004). CONCLUSION: The quality of life in blepharoptosis patients was significantly better after surgical management in the assessment using the VFQ. The outcomes emphasize the need for of using questionnaires designed specifically for this disease. PMID- 17119729 TI - [Study on choroidal melanoma at the Ophthalmology Unit of the Regional Hospital of Sao Jose--Santa Catarina]. AB - PURPOSE: To describe the characteristics of patients choroidal melanoma diagnosed and treated at the Ophthalmology Unit of the Regional Hospital of Sao Jose - Santa Catarina, and to show the results. METHODS: Analysis of 24 medical records of patients with a diagnosis of malignant choroidal melanoma from, March 1st 1997 to July 21st 2003, regarding: age, gender, race, symptoms and duration, diagnostic methods, performed treatment, size and type of tumor. RESULTS: There was no difference regarding gender, and the average age of patients was about 57.2 +/- 16.56 years. The tumor predominated in white people (83.33%) and low visual acuity was the most frequent symptom (83.33%). The average time from the beginning of symptomalogy up to the initial treatment was 152.12+/-114.00 days. Indirect ophtalmoscopy and ultrasound were performed in all the patients in order to diagnose the tumor. All patients underwent enucleation, because they showed large tumors (greatest basal diameter higher than 16 mm at to USG). The most frequent type was that of fusiform cells (58.5%) and the average size of the tumors was 11.8+/-4.5 mm basal diameter. Two patients presented systemic metastasis. CONCLUSIONS: All patients were diagnosed when the tumor was already advanced, making the visual and systemic prognostic worse and reducing the therapeutic options, and therefore were enucleated. PMID- 17119730 TI - [Keratoconus special soft contact lens fitting]. AB - PURPOSE: To evaluate the fitting and use of a soft contact lens in keratoconic patients. METHODS: Retrospective study on 80 eyes of 66 patients, fitted with a special soft contact lens for keratoconus, at the Contact Lens Section of UNIFESP and private clinics. Keratoconus was classified according to degrees of disease severity by keratometric pattern. Age, gender, diagnosis, keratometry, visual acuity, spherical equivalent (SE), base curve and clinical indication were recorded. RESULTS: Of 66 patients (80 eyes) with keratoconus the mean age was 29 years, 51.5% were men and 48.5% women. According to the groups: 15.0% were incipient, 53.7% moderate, 26.3% advanced and 5.0% were severe. The majority of the eyes of patients using contact lenses (91.25%) achieved visual acuity better than 20/40. To 88 eyes 58% were tihed with lens with spherical power (mean -5.45 diopters) and 41% with spherocylinder power (from -0.5 to -5.00 cylindrical diopters). The most frequent base curve was 7.6 in 61% of the eyes. The main reasons for this special lens fitting were due to reduced tolerance and poor fitting pattern achieved with other lenses. CONCLUSION: The special soft contact lens is useful in fitting difficult keratoconic patients by offering comfort and improving visual rehabilitation that may allow more patients to postpone the need for corneal transplant. PMID- 17119731 TI - [Evaluation of the cut quality of Masyk microkeratome in obtaining corneal flap from human eyes of an eye bank]. AB - PURPOSE: To evaluate the cut quality of Masyk microkeratome in obtaining a corneal flap from human eyes of Eye Banks. METHODS: Prospective study with 20 human eyes from Eye Banks. All flaps were programmed to obtain 160 microm thickness and 9.5 mm diameter. The thicknesses were calculated with P55 pachymeter (Paradigm, USA), and diameter with compass. RESULTS: No complications were observed during the use of the microkeratome. The central corneal thickness average was 160.34+/-5.10 microm and range from 149 to 181 microm. The vertical diameter average was 9.64+/-0.16 mm and range from 9.30 to 9.85 mm. CONCLUSION: Masyk microkeratome showed to be effective for obtention of corneal flap with appropriate thickness and diameter from human corneas of Eye Banks. PMID- 17119732 TI - [Tear antioxidant potential in young adults]. AB - PURPOSE: To study the antioxidant status of human tears, stimulated by onion fumes and the possible influence of the life habits thereon were measured. METHODS: Subjects were ten healthy young adults, who answered a questionnaire about smoking, alcohol ingestion, fruit, vegetable, cereal, and vitamin intake and/or intake of other drugs, and physical exercise habits. Chemoluminescensce of luminol was used to analyze the total reactive antioxidant potential (TRAP), inhibition of piragollol was used to measure superoxide dismutase (SOD) and absorbance of H2O2 at 240 nm was used to identify catalase. RESULTS: Mean +/- SD value for total reactive antioxidant potential was 33.8+/-11.5 microM and for superoxide dismutase 10.8+/-1.9 U/mL. Catalase was not identified. Regular exercise was associated with increased total reactive antioxidant potential values (p=0.021), with a difference of 18.6 microM between individuals who exercise at least once a week and sedentary individuals. Male gender and total reactive antioxidant potential values were statistically associated (p=0.013), with a difference of 16.3 microM between genders. There was an association between smoking and increased superoxide dismutase values (p=0.041), with a difference of 3.3 U/mL between smokers of more than five cigarettes/day and non smokers. Also, vitamin C intake and superoxide dismutase values were associated (p=0.018); the difference for vitamin C takers was 3.3 U/mL. CONCLUSION: Reflex tearing antioxidants were measurable in healthy young adults, and different variables apparently influenced their values. PMID- 17119733 TI - [Functional outcome of visually handicapped children cared for at the Department of Visual Stimulation--"Fundacao Altino Ventura"]. AB - PURPOSE: To analyze the functional outcome of visually handicapped children. METHODS: Twenty-seven children were evaluated using the standard functional PEDI test. Seventeen who attended school at "Escola Municipal Alto do Maracana", Recife - Pernambuco, Brazil, had normal visual acuity and the other ten had visual handicap, and were cared for at Stimulation Visual Department - "Fundacao Altino Ventura", in the same city. RESULTS: The children who had visual handicap showed significantly worse performance in self-care and mobility than normal children. CONCLUSION: Visual handicap may have limited the performance regarding self-care, mobility, understanding, communication, home works, in the studied group. PMID- 17119734 TI - [Visual evoked potential in leprosy patients]. AB - PURPOSE: To verify the possible alterations in visual evoked potential of leprosy patients. METHODS: Visual evoked potential examinations were performed in 13 Hansen disease patients, 5 of them with the multibacillary form and 8 with the paucibacillary. This was done at the moment of the diagnosis. The control group enrolled 15 health individuals, without Hansen disease. RESULTS: The measurement result varied from 102.0 to 120.5 ms, with a mean of 110.1+/-5.7 ms. In the multibacillary form, the values varied from 109.0 to 120.0 ms, mean 111.1+/-5.4. In the paucibacillary, the values varied from 102.0 to 120.5 ms, with a mean of 109.5+/-6.1 ms. The latency values were significantly greater in Hansen disease patients (p<0.0001), even if multibacillary and paucibacillary forms were compared separately. However, there was no significant difference when the pauci- and multibacillary groups were compared. CONCLUSION: The latency values were significantly greater in leprosy patients. A recommendation can be made for these patients, which is to be submitted to early visual evoked potential as a rule in order to investigate ocular complications, as well as to prevent damages. PMID- 17119735 TI - [Corneal thickness progression from the thinnest point to the limbus: study based on a normal and a keratoconus population to create reference values]. AB - PURPOSE: To study the variation and progression of the pachymetric values from the thinnest point towards the limbus in normal and keratoconic corneas; to establish reference curves for this parameter. METHODS: One hundred eyes with normal corneas and twenty-five eyes with mild keratoconus (stages I and II - Krumeich) were analyzed using the Orbscan. Concentric circles were drawn on the thinnest point of the cornea with increasing radii from 1 to 7 mm. The average results of each circle were calculated and inserted in an Excel table in order to arrange a progression chart for each case starting on the thinnest point. The SPSS software was also used for statistical analysis. Student's t test was then used to compare the found values. RESULTS: The average values on the thinnest point in normal patients was 511.6 microm (standard deviation 30.6). The average values of the thinnest point in patients with keratoconus was 424.4 microm (standard deviation 56.57). Statistically significant different values were reported (p<0.01) for all circles, and for 6 and 7 mm radii p=0.01. CONCLUSION: There is a greater pachymetric variability in patients with keratoconus. Keratoconic corneas also have a faster progression of pachymetric values than healthy eyes. Pachymetric progression complements traditional single point evaluation of corneal thickness and must be considered for the screening at refractive candidates. This parameter may represent an indirect index of the biomechanics of corneal tissue but this hypothesis still needs further studies. PMID- 17119736 TI - [New surgical approach in the treatment of incomitant strabismus: case report]. AB - The authors report the utilization of the surgical technique proposed by Scott, in 1994, of recess and resection of the same extraocular muscle in the treatment of incomitant horizontal strabismus. A 22-year-old female presented with incomitant strabismus, with an esotropia of 9delta for distant and 30delta for near. She was submitted to a recess and resection of the right medial rectus. She presented satisfactory results in the long-term, showing that this technique could be part of the therapeutic arsenal for correction of incomitant strabismus. PMID- 17119737 TI - [Optical coherence tomography aspects of Stargardt's disease: case report]. AB - The term fundus flavimaculatus (Stargardt disease) describes a group of inherited macular dystrophies characterized by multiple yellow to yellow-white flecks at the level of the retinal pigment epithelium. The authors describe findings in the patient with Stargardt's disease using optical coherence tomography (OCT), and suggest the examination to be valid as subsidiary method in the study of the characteristics of the retina in Stargardt's disease patients, but studies involving a series of patients should be able to show the most frequent findings in these cases. PMID- 17119738 TI - [Autoimmune optic neuropathy: case report]. AB - We report on a 9-year-old female patient who had bilateral severe visual loss and was treated with oral corticosteroids. Visual improvement occurred in one eye. Nine years later she presented relapse of visual loss in her only seeing eye. Pulse corticosteroid therapy resulted in dramatic visual improvement followed, however, by progressive and complete visual loss as soon as the corticosteroid was tapered. Repeat treatment did not result in visual improvement. Clinical and laboratory investigation failed to find a systemic disease but the patient had positive antinuclear (1/640), anti-Ro and anti-La antibodies. Autoimmune optic neuropathy is a rare condition that may mimic an idiopathic optic neuritis and is characterized by acute visual loss, without systemic disease but with laboratory evidence of an autoimmune disorder, usually a positive ANA. A skin biopsy usually shows evidence of vasculitis. This condition should be treated aggressively, with corticosteroids and immunosuppressant, since the visual involvement is usually worse than that of idiopathic/desmyelinating optic neuritis. PMID- 17119739 TI - [Opaque intraocular lens in intractable diplopia: case report]. AB - Intractable diplopia has been described in many situations but poor results are the rule with standard treatment modalities. The authors report a case of a woman with long-standing strabismus and diplopia who failed to improve following surgery, prism, and occlusive spectacles or contact lenses. Then, she was submitted to phacoemulsification and opaque intraocular lens implantation, which successfully neutralized diplopia. PMID- 17119740 TI - [Central vein occlusion in a patient using interferon and ribavirin: case report]. AB - Interferon and ribavirin are medications widely used in the treatment of some systemic diseases, mainly hepatitis C. Ribavirin when associated with interferon increases the rate of success of this treatment. There are about 170 million patients with chronic hepatitis C in the world, many in use of these medications. The classic associated retinopathy is described as cotton wool exudates and hemorrhages. Since the first reports, several different ocular disturbances were described in association with interferon. The present case shows a patient whose right eye presented with central retinal vein occlusion and whose left eye presented the typical findings of hemorrhages; prompt resolution after the medications were discontinued. PMID- 17119741 TI - [Non-penetrating filtering surgery: concept, technique and results]. AB - Recently, many discussions occurred concerning non-penetrating filtering surgery. Viscocanalostomy and deep sclerectomy with external trabeculectomy are the most practiced techniques. Their goal is to reduce intraocular pressure by enhancing the natural aqueous outflow, while reducing its resistance. Both techniques involve the removal of a deep scleral flap, the external wall of Schlemm's canal and corneal stroma behind the anterior trabecular meshwork and Descemet membrane, thus creating a scleral lake, where the aqueous humor remains until it is absorbed in many different ways. In viscocanalostomy, a high-molecular viscoelastic substance is injected into the ostia of Schlemm's canal in order to enlarge it and its collectors channels. In deep sclerectomy with external trabeculectomy, the inner wall of Schlemm's canal is removed with adjacent trabecular layers. Studies, both retrospective and prospective, demonstrated similar tensional results when compared to classical trabeculectomy, with fewer complications and better visual recovery after non-penetrating procedure. The aim of this study is to expose the non-penetrating surgery techniques, mechanism of action and results based on the analysis of published literature. PMID- 17119742 TI - [Perceptions of ophthalmologists regarding teaching models applied to contact lenses]. AB - PURPOSE: To identify perceptions regarding teaching models applied to contact lenses. METHODS: A transversal study in a readily accessible sample consisting of ophthalmologists (n=39) was performed by applying a questionnaire. RESULTS: The sample consisted of 39 ophthalmologists, 51.3% males and 48.7% females. Regarding frequency prior to courses on contact lenses, 28.2% reported only one course; 25.6% two courses; 5.1% three courses; 2.6% five courses and 2.6% more than 10 courses. 23.1% never attended contact lens courses and 12.8% did not answer the questionnaire. On comparison of previous knowledge with that acquired by the offered course, 59.0% declared that they increased their knowledge a lot; 33.3% reported median increase; 2.6% little increase and 5.1% did not answer. As concerns training in contact lenses, 66.7% answered that they never had any training; 30.7% answered affirmatively and 2.6% did not answer. Among those who had training, 75% declared it was good; 16.7%, very good and 8.3%, regular. CONCLUSION: The offered teaching model for contact lenses was considered very valuable, increasing a great deal of knowledge in most subjects. PMID- 17119744 TI - An alternative antimonial schedule to be used in cutaneous leishmaniasis when high doses of antimony are undesirable. AB - Despite more than half a century of use in leishmaniasis, antimony therapy still presents serious problems concerning dosage and toxicity. Low and high doses have been shown to be equally effective. In this paper, the feasibility of injecting one ampoule of meglumine antimoniate intramuscularly every other day until clinical cure is demonstrated, while studying a series of 40 cutaneous leishmaniasis cases. Total dose used varied from 1,822.5 to 12,150 mg of pentavalent antimony and total time of treatment varied from 3 to 10 weeks, with 86% efficacy. Thirty-six out of the 40 patients are still on follow-up with a mean time of 10.7 +/- 7 months and a median of 9 months. No relapse or mucosal lesions have been noted so far. The schedule showed good tolerance and easy application and its efficacy was comparable to the officially recommended WHO schedule. Therefore, such a schedule represents a valuable alternative for the cases with high toxicicity to antimony or daily injections are an obstacle to the treatment. PMID- 17119745 TI - [Periodicity of oviposition of females of Aedes aegypti (Linnaeus, 1762) (Diptera: Culicidae) in laboratory and field]. AB - The object of this work was to determine of gonotrophic diel pattern of female Aedes aegypti in laboratory and field conditions. Three day-old female mosquitoes were the fed on chicken blood and transferred to bioassay cages. Four oviposition substrates were offered: paper sulfite, filter, butter and towel. The results showed that filter paper received a significantly higher (40.4%) percentage of deposited eggs than the other oviposition substrates. After their first blood meal, females started to oviposit on the 3rd model day; 35.7% of the total number of eggs deposited. The oviposition diel patterns of females were observed every two hours during the photoperiod in the laboratory and in the field. In the laboratory, the periodicity of oviposition showed that the highest egg deposition occurred during the 9th- 12th h of photophase and 1st - 2nd h of scotophase. In the field, the highest egg deposition occurred during the 9th - 12th h of photophase and 1st - 4th h of scotophase. These results point out that Aedes aegypti showed an oviposition periodicity pattern that can subsidize monitoring and or control of vector insect. itis suggested that ovitraps should be placed in the field during the morning hours since the captures occur during afternoon. PMID- 17119746 TI - [Focus of canine heartworm disease in Marajo Island, North of Brazil: A risk factor for human health]. AB - The occurrence of human pulmonary dirofilariasis maintains a relation with the prevalence of Dirofilaria immitis infection in the canine population. Several mosquito species are vectors of this nematode. Canine blood samples collected in Pingo d'Agua and Uniao villages, Salvaterra municipality (Marajo Island, Para), in June, 2004 (n = 34) and April, 2005 (N = 90) were analyzed. Parasitological and immunological (ELISA--kit SNAP(R) 3DX Biobrasil) diagnoses were compared following the examination of 34 samples. The prevalence in the population (N = 90) was evaluated by means of ELISA. ELISA revealed more positive samples (25/34; 73.5%) than thick smears (23/34, 67.6%) or Knott (21/34, 61.8%), but the differences were not significant (p > 0.05). The frequency of D. immitis infection was 58% in dogs ranging from 0-2 years old, whereas in older dogs it was 100%. The prevalence of canine dirofilariasis was high in Pingo d'Agua and Vila Uniao (53.5%), indicating the risk of parasite transmission to the people in this area. PMID- 17119747 TI - [Spatial distribution of Lutzomyia longipalpis (Lutz & Neiva, 1912) and Lutzomyia cruzi (Mangabeira, 1938) in the State of Mato Grosso]. AB - Visceral leishmaniasis is currently considered an emergent and reemergent disease, in agricultural and urban zones, both in domestic and peridomestic areas. The objective of this work was to verify the spatial distribution of Lutzomyia longipalpis and Lutzomyia cruzi in Mato Grosso State. Data from 1996 to 2004 was collected by the Entomology Laboratory, whose captures were carried with a CDC light trap. Sixty-eight of the 139 cities in the state have been studied. Lutzomyia longipalpis and Lutzomyia cruzi occurred in 23 and 22 cities, respectively. The results demonstrate the extensive occurrence of Lutzomyia longipalpis in areas with different vegetation types: savanna, transition and Amazon forest. Lutzomyia cruzi occurred mainly in cities with areas of marshland and savanna. Verification of the distribution of the vector populations in the state and their preferential vegetation type, provides an indication of vulnerable and/or receptive areas for disease transmission. PMID- 17119748 TI - Phlebotominae sand flies associated with a tegumentary leishmaniasis outbreak, Tucuman Province, Argentina. AB - The distribution of sand flies and cases of tegumentary leishmaniasis in the area surrounding JB Alberd city, and the proximities of Catamarca province were studied, after an increase of reported cases from JB Alberdi, Tucuman province, in 2003. Of 14 confirmed cases, 57% were females and 57% were less than 15 years old, suggesting peridomestic transmission. However, 86% of them lived close to the Marapa river forest gallery and related wooded areas. Over 1,013 sand flies were collected; Lutzomyia neivai (Pinto, 1926) was prevalent at all the sites (92.3%), while Lutzomyia migonei (Franca, 1920) (6.7%) and Lu. cortelezzii (Brethes, 1923) (1%) were also found. The spatial distribution of Lu. neivai overlapped that of the cases, with higher abundance in microfocal hot spots close to the river in stable vegetated habitats or modified habitats with shadow and animal blood sources. The cumulative outcome of anthropic, ecological and climatic factors could have contributed to the onset of the outbreak. PMID- 17119749 TI - [Feeding sources evaluation of Panstrongylus lutzi (Neiva & Pinto, 1923) (Hemiptera: Reduviidae: Triatominae) in the State of Ceara]. AB - The authors used precipitin technique to detect the feeding sources of Panstrongylus lutzi (Neiva & Pinto, 1923) in 20 municipalities of Ceara State, Brazil. The results detected the presence of blood from eight different blood sources and mixed feedings, demonstrating that Panstrongylus lutzi is an eclectic species. High infection rates for Trypanosoma like-cruzi were detected. PMID- 17119750 TI - [Geographical expansion of visceral leishmaniasis in the State of Pernambuco]. AB - This study aimed to demonstrate the geographical expansion of visceral leishmaniasis in Pernambuco State, Brazil. With data on the number of visceral leishmaniasis cases notified from 1990 to 2001, maps showing the biannual evolution of the geographical distribution of the disease per municipality were elaborated. A map of the cumulative geographical distribution of the cases registered during the whole period was also constructed. From 1990 to 2001, 1,737 cases of visceral leishmaniasis were notified in Pernambuco. In 1990, 15.2% (n = 28) of the municipalities notified cases of the disease. In contrast, this percentage increased to 78.3% (n = 144), over an eleven-year period. The map of cumulative geographical distribution during the whole period shows a notable concentration of cases in Agreste and Sertao. A notable geographical expansion of visceral leishmaniasis in Pernambuco also occurred, probably reflecting the low impact of the current control measures and, possibly, an improvement in the notification system. PMID- 17119751 TI - Dynamics and kinetics of natural killer cell cytotoxicity in human malaria as evaluated by a novel stepwise cytotoxicity assay. AB - Malaria causes important functional alterations of the immune system, but several of them are poorly defined. To evaluate thoroughly the natural killer cell cytotoxicity in patients with malaria, we developed a technique capable to assess both the dynamics and the kinetics of the process. For the kinetics assay, human peripheral blood mononuclear cells were previously incubated with K562 cells and kept in agarose medium, while for the dynamics assay both cells were maintained in suspension. NK activity from patients with vivax malaria presented a kinetics profile faster than those with falciparum malaria. NK cytotoxicity positively correlated with parasitemia in falciparum malaria. The dynamics of NK cytotoxicity of healthy individuals was elevated at the beginning of the process and then significantly decreased. In contrast, malaria patients presented successive peaks of NK activity. Our results confirmed the occurrence of alteration in NK cell function during malaria, and added new data about the NK cytotoxicity process. PMID- 17119752 TI - [Hope Clubs as adjunct therapeutic measure in bancroftian filariasis endemic areas]. AB - In 1997 the World Health Organization announced an ambitious project called the Global Program to Eliminate Lymphatic Filariasis, as a Public Health Problem. The program is based on two pillars: interruption of transmission and morbidity control. Experience in Recife, Brazil, an endemic area for bancroftian filariasis, showed that an innovative approach called Hope Clubs, can equip lymphedema patients with the skills, motivation, and enthusiasm to sustain effective, low-cost and convenient self-care to prevent acute skin bacterial episodes and milky urine in the case of chyluria carriers. They feel they are not alone, they regain their potential for productive work and are able to amplify these activities throughout filariasis-endemic communities. PMID- 17119753 TI - [Notes about of Trypanosoma cruzi and yours bio-ecology characteristics with agents of the transmission by meals]. AB - Experimental evidence and field observations concerning the oral transmission of Trypanosoma cruzi to human beings and mammalian species are reviewed, confirming its unquestionable occurrence, mainly in the enzootic cycle of the parasite. Several types of foods and carriers of the flagellate have been involved in this transmission route, where the human cases are generally related to infected triatomines and/or natural reservoirs in the proximities of the event. The penetration points of the parasite (mouth, esophagus, stomach and intestinal mucosae) are reviewed, as well as the general anatomic and pathological processes, such as hemorrhagic phenomena, mesenteritis and interstitial hepatitis. In particular, different aspects of the biology and biochemistry of the parasite are analyzed, especially in terms of its epidemiology and the possibilities of the prevention of the oral transmission of Chagas' disease. PMID- 17119754 TI - Successful therapeutic response of resistant cases of mucocutaneous leishmaniasis to a very low dose of antimony. AB - Two mucocutaneous leishmaniasis cases resistant to therapy are reported here. After the failure of initial therapies (antimony, amphotericin B and/or pentamidine) patients received a low-dose schedule: one ampoule of meglumine antimoniate (405 mg of pentavalent antimony [Sb v]) by intramuscular injection, three times a week until complete healing of the lesions. One patient was cured with a total of 30 ampoules in 10 weeks and the other received 36 ampoules in 12 weeks. Both remain clinically cured after one year of follow-up. PMID- 17119755 TI - Schistosomiasis mansoni and severe gastrointestinal cytomegalovirus disease in a patient with acquired immunodeficiency syndrome. AB - The behavior of the Schistosoma mansoni infection in patients with AIDS has not been explored. The case of a young woman with schistosomiasis mansoni, AIDS, and cytomegalovirus disease is reported. The authors suggest that the helminth was not a bystander in this case, or rather, by interfering with the host's immune response, it set the stage for the development and/or aggravation of the viral infection. PMID- 17119756 TI - [First record of scorpionism caused by Tityus adrianoi Lourenco (Scorpiones: Buthidae)]. AB - Three cases of scorpionism caused by Tityus adrianoi Lourenco occurring in the cities of Belo Horizonte and Curvelo, Minas Gerais State, Brazil, are presented. The patients were treated on the same day as the accident and in all cases, only local pain was reported, which varied from low to intense, without posterior complications. All three cases were classified as mild. PMID- 17119757 TI - Assessment of chemiluminescence and PCR effectiveness in relation to conventional serological tests for the diagnosis of Chagas' disease. AB - While testing 414 sera for the diagnosis of Chagas' disease, the conventional reactions of indirect hemagglutination, indirect immunofluorescence and the immunosorbent assay showed a sensitivity of 95.7%, 100% and 98.2% and a specificity of 98%, 98% and 96.4%, respectively, and an excellent association using Fisher's exact test. Chemiluminescence presented 100% sensitivity and 89.6% specificity, while PCR showed 100% specificity and 1.2% sensitivity. It is believed that the three conventional serological reactions are still adequate for diagnosing Chagas' disease. PMID- 17119758 TI - [Occurrence of Entamoeba histolytica/Entamoeba dispar in ambulatory patients of Recife, PE]. AB - The objective this study was to determine the occurrence of the species Entamoeba histolytica/Entamoeba dispar in clinical samples of ambulatory patients in Pernambuco. A specific assay for Entamoeba histolytica was used in this study, which identified no positive fecal samples among the 213 patients. These results confirm that E. dispar is the dominant species in Pernambuco State. PMID- 17119759 TI - Rapid progressive dementia associated with neurosyphilis. PMID- 17119760 TI - [Recommendations for diagnosis, treatment and follow-up of the Trypanosoma cruzi: human immunodeficiency virus co-infection]. PMID- 17119761 TI - [Heonir de Jesus Pereira da Rocha (*1930-2005)]. PMID- 17119762 TI - The uncertain future. PMID- 17119763 TI - Vocal assessment in patients submited to CO2 laser cordectomy. AB - AIM: To evaluate voice outcomes in patients with early glottic carcinoma treated by CO2 laser cordectomy. METHOD: 15 patients with glottic Tis and T1 squamous cell carcinoma treated with CO2 laser were analyzed. The assessment consisted of perceptual voice analysis, objective voice evaluation and video-laryngo stroboscopic exam. In addition, patients rated their voices and completed the Voice related Quality of Life (VR-QOL) questionnaire. The results were compared with those obtained in a matched control group. RESULTS: Most of the patients presented some degree of hoarseness on perceptual voice analysis, mainly rough and breathy voices. Their acoustic evaluation compared with the control group showed a small increase in fundamental frequency, but with no statistically significant difference, and the values of jitter, shimmer and noise to harmonic ratio were worse and statistically significant. As regards to videolaryngo stroboscopic findings, better results were achieved in the less extensive resection group. Patients have had minimal repercussion in their life quality in respect to voice. CONCLUSIONS: In spite of voice alterations in patients submitted to cordectomy by CO2 laser, functional results are acceptable, with minimal repercussion in their quality of life. PMID- 17119764 TI - Endoscopic anatomy of the anterior ethmoidal artery: a cadaveric dissection study. AB - INTRODUCTION: The anterior ethmoidal artery (AEA) is an important point of anatomical reference in order to locate the frontal sinus and the skull base. Notwithstanding, despite numerous endoscopic studies in cadavers, we still lack an anatomical study on the AEA in the western population. AIM: to determine reference points used to locate the artery, study its relationship with the skull base and its degree of dehiscence, as well as to study intra and inter individual variations. MATERIALS AND METHODS: we dissected the nasal fossae belonging to 25 cadavers. RESULTS: the average intranasal length of the anterior ethmoidal artery was 5.2 mm. The anterior ethmoidal canal presented some degree of dehiscence in 66.7%. The average distance between the artery middle point to the anterior nasal spine was of 61.72 mm (sd=4.18 mm); to the lateral nasal wall (nasal axilla) was of 64.04 mm (sd=4.69 mm); and from the anterior axilla to the middle turbinate was of 21.14 mm (sd=3.25 mm). For all the measures there was no statistically significant measures when both sides were compared (p>0.05). CONCLUSIONS: We concluded that the middle conchae axilla is the most reliable point of reference to locate the AEA. PMID- 17119765 TI - The oral mucosa in leprosy: a clinical and histopathological study. AB - INTRODUCTION: Multibacillary leprosy may involve the oral mucosa, with or without apparent lesions. There are few studies that deal with this issue in the era of multidrug therapy. AIM: To assess the frequency of oral mucosa involvement in multibacillary leprosy patients. PATIENTS AND METHODS: A transversal study with twenty non-treated multibacillary leprosy patients. The patients were treated in Dracena, Sao Paulo, between 2000 and 2002. Clinical examination of the oral mucosa was carried out. All patients were submitted to jugal mucosa, soft palate and tongue biopsies, in altered or in pre-established sites. The cross-sections were stained by techniques of hematoxilin-eosin and Ziehl-Neelsen. Granuloma and alcohol-acid-resistant bacilli findings determined the specific histopathological involvement. RESULTS: The study involved 19 patients with an average of 2.5 years of disease progression. Specific histopathological involvement occurred in the tongue and soft palate of one lepromatous patient with an apparently normal oral mucosa. CONCLUSIONS: (1) Clinical alterations in the oral mucosa does not imply disease involvement, it is necessary to have histopathological confirmation. (2) Apparent specific clinical alterations are rare. (3) The clinically normal oral mucosa can show specific histopathological involvement. PMID- 17119766 TI - Auditory processing assessment in children with obstructive sleep apnea syndrome. AB - INTRODUCTION: The obstructive sleep apnea syndrome (OSAS) is a respiratory disorder that occurs during sleep and it is relatively common in children. AIM: The goal of this paper is to verify if there is a relationship between the obstructive sleep apnea syndrome (OSAS) and auditory processing. METHODS: In order to do that, three groups of children ranging in age from 5 to 11 were studied, including a normal group. Twenty subjects who made up the study group were submitted to ear, nose and throat (ENT) exams and to polysomnography (PSG), and were divided in two groups: GROUP I (RO) comprised of 10 children who presented oral breathing and displayed normal PSG, and GROUP II (SAS) comprised of 10 children who presented oral breathing and displayed abnormal PSG. Their performance was compared to the performance of the third group--GROUP III (REN) comprised of 10 children who did not refer ENT difficulties. All the subjects completed a basic audiometric assessment as well as an auditory processing diagnosis. RESULTS: The analyses of the results revealed a statistically significant difference in ENT exams related to the turbinate and the palatine tonsils. Group II presented a higher incidence of turbinate hypertrophy levels II and III (p<0.001) and palatine tonsils hypertrophy grades III and IV (p 0.007). Regarding the auditory processing assessment, a statistically significant difference (p<0.001) was obtained in the dichotic digits test. Group II performed worse than group III. Also, for the non-verbal sequence memory test, Group II obtained a worse result (p<0.022) than Group I. CONCLUSION: Subjects with OSAS obtained worse results in auditory processing tests. PMID- 17119767 TI - Chronic GVHD: predictive factor for rhinosinusitis in bone marrow transplantation. AB - INTRODUCTION: Bone marrow transplantation (BMT) is a treatment option for hematological diseases and immunodeficiency. It is frequently used today. BMT predisposes patients to upper airway infections and its complications, such as rhinosinusitis (RS). Chemotherapy, radiotherapy, viral infections, antibiotic therapy, graft versus host disease (GVHD) are rhinosinusitis predisposing conditions. AIM: to investigate RS frequency in this population and its relationship to GVHD; to try and establish the best treatment for RS in these patients. METHOD: ENT evaluation of two groups. One group with 35 patients (gI) and another with 24 patients (gII), before and after BMT. They were treated with antibiotics, maxillary sinus punction or endoscopic sinusectomy. RESULTS: none of them had RS before BMT. 42.8% from gI had RS and 34% had GVHD; in the gII, 58% had RS and 25% had GVHD. 49% from both groups had RS and 30.5% had GVHD. There was significantly more RS in chronic GVHD patients. Surgery was used to treat RS in chronic GVHD patients who underwent BMT. CONCLUSION: RS frequency was 49%; GVHD is a predisposing condition to RS; sinusectomy may be necessary to control RS in GVHD patients. PMID- 17119768 TI - Molecular investigation in children candidates and submitted to cochlear implantation. AB - AIM: Recent progresses in molecular biology have been made in the diagnosis of sensorineural hearing loss. The high prevalence of a connexin 26 gene mutation, and its easy identification have made the diagnosis possible. The most frequent gene mutation is called 35delG. The purpose of this study was to evaluate the prevalence of 35delG mutation in children submitted to cochlear implantation who had severe and profound hearing loss previously diagnosed as idiopathic. METHOD: The study was done at the Cochlear Implantation Clinic of the Otolaryngology Department and at the Laboratorio Genetica Humana-CBMEG, UNICAMP-SP. 32 children with severe to profound sensorineural hearing loss were evaluated. The detection of the 35delG mutation was made by a allele-specific PCR, using primers and polymerase chain reaction. RESULTS: 69% had a normal exam, 12% were homozygous for the mutation, 19% of the cases were heterozygous. The 35delG mutation in heterozygous is not a cause of hearing loss. CONCLUSION: The data confirm the high prevalence of the 35delG mutation in nonsyndromic bilateral profound sensorineural hearing loss. It was also possible to diagnose the cause of hearing loss as genetic in a significant percentage of patients. That stresses the importance of the molecular investigation in those cases formerly classified as idiopathic. PMID- 17119769 TI - Standardization of techniques used in facial nerve section and facial movement evaluation in rats. AB - AIM: Standardization of the technique to section the extratemporal facial nerve in rats and creation of a scale to evaluate facial movements in these animals before and after surgery. STUDY DESIGN: Experimental. METHOD: twenty Wistar rats were anesthetized with ketamine xylazine and submitted to sectioning of the facial nerve near its emergence through the mastoid foramen. Eye closure and blinking reflex, vibrissae movement and positioning were observed in all animals and a scale to evaluate these parameters was then created. RESULTS: The facial nerve trunk was found between the tendinous margin of the clavotrapezius muscle and the auricular cartilage. The trunk was proximally sectioned as it exits the mastoid foramen and the stumps were sutured with a 9-0-nylon thread. An evaluation and graduation scale of facial movements, independent for eye and vibrissae, was elaborated, together with a sum of the parameters, as a means to evaluate facial palsy. Absence of eye blinking and closure scored 1; the presence of orbicular muscle contraction, without blinking reflex, scored 2; 50% of eye closure through blinking reflex, scored 3, 75% of closure scored 4. The presence of complete eye closure and blinking reflex scored 5. The absence of movement and posterior position of the vibrissae scored 1; slight shivering and posterior position scored 2; greater shivering and posterior position, scored 3 and normal movement with posterior position, scored 4; symmetrical movement of he vibrissae, with anterior position, scored 5. CONCLUSION: The rat anatomy allows easy access to the extratemporal facial nerve, allowing its sectioning and standardized suture. It was also possible to establish an evaluation and graduation scale of the rat facial movements with facial palsy based on the clinical observation of these animals. PMID- 17119770 TI - Audiometric and vestibular evaluation in women using the hormonal contraceptive method. AB - AIM: To co-relate the use of hormonal contraceptives with positive Auditory and Vestibular alterations. METHODS: medical history taking, audiometric test and vestibular test was applied to 60 women between the ages of 14 and 35 years old, and 30 of these women are on oral hormonal contraceptive consisting of Estrogen and Progesterone (risk group), for 6 months or more, that had no Auditory or Vestibular complaints prior to the usage of hormones; and 30 women that had never used these hormones (control group), with no Auditory or Vestibular complaints. Medical history was used to select the sample. RESULTS: Based on otoneurological findings, through quantitative research, we could see the prevailing Irritatative Peripheral Vestibular Syndrome and tinnitus in the risk group, without audiometric alterations. CONCLUSION: The use of oral contraceptives can provoke functional alterations in the inner ear, specially tinnitus and Irritative Peripheral Vestibular Syndrome in the risk group; but auditory threshold alterations were not evident. PMID- 17119771 TI - OSAS in children: clinical and polysomnographic respiratory profile. AB - Obstructive sleep apnea and hypopnea syndrome in children (osas) has an estimated prevalence of up to 3% and can be associated with neurocognitive and behavioural abnormalities, and also cardiovascular complications. This study may help pediatricians, who are unaware of the problem, to recognize osas. STUDY DESIGN: series of cases. AIM: to describe the clinical characteristics and polysomnographic respiratory findings in a population of children with obstructive sleep apnea and hypopnea syndrome referred to the sleep laboratory from january 2002 up to july 2003. METHODS: we studied 93 patients between 2 and 10 years of age with polysomnographic diagnosis of obstructive sleep apnea and hypopnea syndrome. Age, gender, racial group and questions about the childrens health and sleep related disorders were evaluated. Apnea-hypopnea index, oxyhemoglobin desaturation, and arousal index were evaluated too. RESULTS: males represented 61.3%, With a mean age of 5.2+/-2.1 (Years-old). The complaints that most commonly lead to the exams were snoring in 24.7% And restless sleep in 24.7%. Associated medical conditions frequently reported were allergic rhinitis (98.9%) And adenoid hypertrophy (50.6%). Mild apnea was found in 66%. The mean and sd of spo2 nadir was 89.1+/-3.5% And the mean and sd of the number of arousals was 8.4+/-3.5/Hour of sleep. CONCLUSION: the results suggest the possibility that obstructive sleep apnea and hypopnea syndrome should be suspected in children with allergic diseases and adenoid and tonsil hypertrophy with snoring and restless sleep complaints. PMID- 17119772 TI - Exposure to occupational noise: otoacoustic emissions test alterations. AB - Exposure to occupational noise may cause injuries to the inner ear, and the distortion product otoacoustic emissions (DPOAE) may identify initial auditory alterations, thus assisting NIHL early diagnosis. AIM: The goal of this study was to evaluate DPOAE as a method to diagnose early physiopathological alterations caused by occupational noise exposure. STUDY DESIGN: Transversal. METHODS: 74 workers of the University of Sao Paulo, in the capital city of the State, participated in this investigation. They were divided in two age-matched groups and with tonal audiometric values within the acceptable limits: 37 were exposed to occupational noise and 37 were not exposed. RESULTS: Risk estimates (Odds Ratio) of absent DPOAE was 12 fold higher for the group exposed to occupational noise (CI 95% 3.1-45.9), in the frequencies of 3. 4 and 6 kHz. CONCLUSION: DPOAE may be useful in the identification of physiopathological hearing alterations caused by exposure to occupational noise, even in individuals with tonal audiometric responses within acceptable limits. PMID- 17119773 TI - Correlation between dizziness and impaired glucose metabolism. AB - INTRODUCTION: Impaired glucose metabolism is characterized by conditions of hypo and hyperglycemia. AIM: The objective of the present study was to asses whether or not there is a relationship between impaired glucose metabolism and dizziness. In the clinical laboratory settings, patients were examined using vectoelectronystagmography in association with glycemic levels. METHODS: 33 patients were divided in 3 groups: diabetics; patients with dizziness and a control group. RESULTS: 65% of the patients with dizziness showed impaired glucose metabolism. 40% of the patients with dizziness had alterations in their vectoelectronystagmography results. CONCLUSION: Dizziness is a good indicator of glucose metabolism alterations and these may be a good indicator of alterations in vectoelectronystagmography responses. The study of glycemic levels after glucose overexposure is a good prognosis factor to evaluate dizziness and shows the same results as insulin level studies after glucose overexposure. PMID- 17119774 TI - CT-Scan sheep and human inner ear morphometric comparison. AB - INTRODUCTION: Studies about the use of sheep in surgical training and experimental otologic surgery are rare. This study intends to contribute to the knowledge on this field. AIM: To study sheep inner ear structures using computerized tomography and serial cross-sections to collect more accurate morphometric data to compare sheep and human ears. MATERIAL AND METHODS: This descriptive study compared the inner structures of sheep and human ears. Measurements were made using computerized tomography, and they were stored in a DICOM compact disc for later analysis and manipulation, with a program used for medical image analysis (Osiris 4.16). RESULTS: Mean measures for sheep and human ears were found to be similar in this morphological study. Most structures (10 out of 15) maintained the 2/3 ratio of sheep to human ear. CONCLUSION: The results of this morphometric study of sheep ear are an important contribution to the development of an animal model to be used for surgical training and experimental otologic surgeries. PMID- 17119775 TI - Intraoral and transcutaneous cervical ultrasound in the differential diagnosis of peritonsillar cellulitis and abscesses. AB - AIMS: The objective of the present study was to determine the specificity, sensitivity and accuracy of intraoral and transcutaneous ultrasound (US) in the diagnosis of peritonsillar cellulitis and abscess. STUDY DESIGN: Clinical Prospective. MATERIAL AND METHODS: Thirty nine patients were seen at the otorhinolaryngology emergency department of the University Hospital, of the School of Medicine, University of Sao Paulo, with a clinical diagnosis of peritonsillar cellulitis or abscess. After initial evaluation, all patients were submitted to intraoral and transcutaneous US. RESULTS: Intraoral US was performed on 35 cases and its sensitivity was of 95.2%, the specificity was of 78.5% and the accuracy was of 86.9%. Transcutaneous US was feasible in all 39 patients and diagnosed peritonsillar abscess in 53.8%. There were 5 false-negatives and 1 false-positive result, sensitivity was 80%, specificity was 92.8% and accuracy was 84.5%. CONCLUSION: Intraoral US was quite sensitive in the diagnosis of peritonsillar abscesses when performed by an experienced radiologist. Specificity was higher for transcutaneous US compared to intraoral US. However, when transcutaneous US was performed in patients with trismus, it was able to diagnose all peritonsillar abscesses, since they were large collections which are common in patients with trismus. These exams showed similar accuracy. PMID- 17119776 TI - Lymph node reactivity and microvessel density in neck metastases of unknown primary squamous cell carcinoma. AB - BACKGROUND: Neoangiogenesis and the immune response are important mechanisms in metastasis development. AIM: to evaluate lymph node reactivity and microvessel density in neck metastasis of occult primary squamous cell carcinoma considering their histological and clinical variables. STUDY DESIGN: retrospesctive case series. METHOD: 19 patients with neck metastasis of occult primary squamous cell carcinoma who underwent neck dissection between 1983 and 2000 were selected. The lymph nodes were reevaluated on the type of reactivity in both the cortical and paracortical areas, and the metastasis were assessed as to grade, desmoplasia, necrosis and microvessel density (CD34). The relationship between histological and clinical variables was evaluated. RESULTS: the median microvessel density was 91 vessels/mm2, varying from 28 to 145. Paracortical hyperplasia was more common in patients below 55 years of age (90%x44%, p=0.05), but there was no relationship between reactivity patterns and microvessel density with prognosis. The disease-free survival was 52% in 3 years, being similar in both groups, with higher or lower microvessel densities. CONCLUSION: microvessel density in neck metastasis of occult primary squamous cell carcinoma had a great individual variability. It wasnt possible to establish the relationship between microvessel density and the clinical or histological variables studied. PMID- 17119777 TI - Circumstances and consequences of falls in elderly people with vestibular disorder. AB - AIM: To investigate the circumstances and consequences of falls in the chronically dizzy elderly and to correlate them with the number of falls (one/two and more). METHOD: Transversal descriptive analytic study with 64 patients aged 65 or over, with history of falls and diagnostic of chronic vestibular dysfunction. We performed a descriptive analysis and Chi-Square test (x2<0.05). RESULTS: The sample was constituted by a female majority (76.6%) with a mean age of 73.62+/-5.69 years. The vestibular examination showed peripheral vestibulopathy in 81.5% of the cases and the most prevalent diagnostic hypothesis were benign paroxysmal positional vertigo (43.8%) and metabolic inner ear disease (42.2%). Recurrent falls were seen in 35 elderly (53.1%). In relation to the last fall, 39.1% of the patients had fallen in their homes, 51.6% of them occurred during the morning, 51.6% with some propulsion mechanism, 53.1% when walking, 25.0% caused by dizziness and 23.4% by stumbling. Activity restriction was significantly greater in patients that have already had two and more falls, when compared with those who had fallen only once (p=0.031). We found a significant association between the number of falls and their causes (p<0.001). Falls that have happened by slipping were more frequent in the elderly that reported one fall (p=0.0265) and falls that had happened because of dizziness were more frequent in the elderly that complained of two or more falls (p=0.0012). CONCLUSION: Fear and tendency to fall are referred by the majority of chronically dizzy elderly. Fall are more frequent in the morning, in the home and during walking. The propulsion direction is mentioned by half of the elderly and the most common cause for falls are dizziness and stumbling. The number of falls is significantly associated with activity restrictions after the last fall and with the causes for falling (slipping and dizziness). PMID- 17119778 TI - Prevalence and factors related to mouth breathing in school children at the Santo Amaro project-Recife, 2005. AB - AIM: To determine the prevalence of mouth breathing children at the santo amaro project/ esef/ upe, and study their main facial and behavior alterations. STUDY DESIGN: transversal study. MATERIAL AND METHODS: there were 150 children in the sample, with ages ranging from 8 to 10 years. Data was collected by means of a questionnaire and clinical examinations. As for their breathing assessment, two tests were carried out: test 1- breath steam against a mirror; and test 2 -water remains in the mouth with lips closed for 3 minutes. RESULTS: mouth breathing prevalence was of 53.3%. There was no significant difference between gender, age and type of breathing. Facial alterations were:incomplete lip closure ( 58.8%X5,7%), fallen eyes ( 40.0%X1.4%), High palate ( 38.8%X2.9%), Anterior open bite ( 60.0% Versus 30.0%), Hypotonic lips ( 3.8%X0.0%), Circles under the eyes (97.5% Versus 77.1%). CONCLUSION: high mouth breathing prevalence without significant statistical difference between genders,age and type of mouth breathing. There was no association between behavior characteristics and type of breathing. There were significant differences between physical traits and breathing pattern. PMID- 17119779 TI - Pharmaceutical excipients and the information on drug labels. AB - AIM: to evaluate the presence of preservatives, dyes, sweeteners and flavouring substances in 73 pharmaceutical preparations of 35 medicines for oral administration, according to drug labeling information about the excipients. METHODS: 35 medications were selected, both over-the-counter and prescription drugs, marketed in Brazil. The sample included: analgesic/antipyretic, antimicrobial, mucoregulatory, cough and cold, decongestant, antihistamine, bronchodilator, corticosteroid, antiinflammatory and vitamin medications. We collected data on 73 preparations of these drugs, according to drug labeling information regarding preservatives, dyes, sweeteners and flavourings. RESULTS: Methylparaben and propylparaben were the most common preservatives found (43% and 35.6% respectively). The most common sweeteners were: sucrose (sugar) (53.4%), sodium saccharin (38.3%) and sorbitol (36.9%). Twenty-one medicines (28,7%) contained two sweeteners. Colourless medicines predominated (43.8%), followed by those with sunset yellow dye (FD&C yellow no. 6) (15%). Five products (6.8%) contained more than one colour agent. Tartrazine (FD&C yellow no. 5) was present in seven preparations (9.5%). Fruit was the most common flavouring found (83%). Labelings of drugs which contained sugar frequently omitted its exact concentration (77%). Of the four labelings of medicines which contained aspartame, two did not warn patients regarding phenylketonuria. CONCLUSION: Omission and inaccuracy of drug labeling information on pharmaceutical excipients may expose susceptible individuals to adverse reactions caused by preservatives and dyes. Complications of inadvertent intake of sugar-containing medicines by diabetics, or aspartame intake by patients with phenylketonuria may also occur. PMID- 17119780 TI - Oculomotor evaluation in patients with peripheral vestibular dysfunction. AB - AIM: To assess whether or not the parameters of fixed and randomized saccadic movements, of pendular tracking and of optokinetic nystagmus in the digital vectonystagmography may show abnormalities in patients with possible diagnosis of peripheral vestibular dysfunction. METHOD: 60 patients with dizziness of peripheral vestibular origin, from 12 to 82 years of age, males and females, were evaluated in the Universidade Federal de Sao Paulo. Ocular movement parameter findings were compared to a normal pattern. RESULTS: Fixed saccadic movements were altered in 100% of the cases as to latency, and in 35.0% of the cases as to speed; the randomized saccadic movements were altered in 100% of the cases as to latency, in 78.3% as to precision, and in 1.7% as to speed; the pendular tracking showed a gain alteration in the frequencies of 0.1 Hz in 15% of the cases, 0.2 Hz in 21.7%, and 0.4 Hz in 13.3%; the optokinetic nystagmus showed an alteration of the angular speed in the slow component in 1.7% of the cases, and in gain in 5.0%. CONCLUSION: Fixed saccadic movement latency and speed, randomized saccadic movement latency, precision and speed, pendular tracking gain, slow component angular speed, and optokinetic nystagmus gain in the digital vectonystagmography may show abnormalities in patients with possible diagnosis of peripheral vestibular dysfunction. PMID- 17119781 TI - Superior Canal Dehiscence Syndrome. AB - The Superior Canal Dehiscence Syndrome (SCDS) was first reported by Minor at. Al. (1998), and has been characterized by vertigo and vertical-torsional eye movements related to loud sounds or stimuli that change middle ear or intracranial pressure. Hearing loss, for the most part with conductive patterns on audiometry, may be present in this syndrome. We performed a literature survey in order to to present symptoms, signs, diagnostic and therapeutic approaches to the SCDS, also aiming at stressing the great importance of including this syndrome among the tractable cause of vertigo. We should emphasize that this is a recent issue, still unknown by some specialists. The Correct SCDS diagnosis, besides enabling patient treatment, precludes misdiagnosis and inadequate therapeutic approaches. PMID- 17119782 TI - Burning mouth syndrome: etiology. AB - The Burning Mouth Syndrome (BMS) is an oral mucosa pain--with or without inflammatory signs--without any specific lesion. It is mostly observed in women aged 40-60 years. This pain feels like a moderate/severe burning, and it occurs more frequently on the tongue, but it may also be felt at the gingiva, lips and jugal mucosa. It may worsen during the day, during stress and fatigue, when the patient speaks too much, or through eating of spicy/hot foods. The burning can be diminished with cold food, work and leisure. The goal of this review article is to consider possible BMS etiologies and join them in 4 groups to be better studied: local, systemic, emotional and idiopathic causes of pain. Knowing the different diagnoses of this syndrome, we can establish a protocol to manage these patients. Within the local pain group, we must investigate dental, allergic and infectious causes. Concerning systemic causes we need to look for connective tissue diseases, endocrine disorders, neurological diseases, nutritional deficits and salivary glands alterations that result in xerostomia. BMS etiology may be of difficult diagnosis, many times showing more than one cause for oral pain. A detailed interview, general physical examination, oral cavity and oropharynx inspection, and lab exams are essential to avoid a try and error treatment for these patients. PMID- 17119783 TI - Laryngeal dystonia: case report and treatment with botulinum toxin. AB - Laryngeal dystonia or spasmodic dysphonia is characterized by involuntary and inappropriate spasms of vocal muscles, having the adductor type as the most common one. It is characterized by strain-strangled voice with pitch breaks. Diagnosis is made by means of videolaryngostroboscopic exam. The treatment of choice is done with botulinum toxin directly injected in the muscles responsible for the mismatched movement. The aim of this study is to report on an adductor- type dysphonia patient and to discuss the advantages and observations about this treatment reported in the literature. PMID- 17119784 TI - Bilateral frontal sinus mucocele. PMID- 17119785 TI - Automastoidectomy. PMID- 17119786 TI - Perilymphatic hyperension. PMID- 17119787 TI - Foreign body in nasopharynx: an accidental radiological finding. PMID- 17119788 TI - Mutation analysis of CACNA1A and ATP1A2 genes in Brazilian FHM families. AB - Familial hemiplegic migraine (FHM) is a rare autosomal dominant form of migraine with aura. This disease has been associated with missense mutations in the CACNA1A and ATP1A2 genes. The aim of this study was to identify whether CACNA1A and ATP1A2 are or not related to Brazilian FHM. Here we screened four Brazilian FHM families (total of 26 individuals--13 affected and 13 asymptomatic or normal) for mutations in both genes. We found an amino acid change in a member of family FHM-D (Arg2206Gly). However since this alteration is not present in all affected individuals and is present in one asymptomatic individual it should be considered a polymorphism. Further studies with additional families will be necessary to reveal the importance of both CACNA1A and ATP1A2 genes on the pathogeneses of FHM in Brazil and to test the third gene (SCN1A) in these FHM families. PMID- 17119789 TI - Event related potentials during the visual discrimination of words and pseudowords by children. AB - Event related potentials (ERPs) in reading were studied in children in a word and pseudoword discriminating task. Seventy-nine children (9 to 11 year old), all with no elements suggesting brain injury and with school performance compatible with their age were studied. The ERP were registered as there were presented, visually, successively and in a random manner, 100 words and 100 pseudowords. For each stimulus the child pressed a key corresponding to the discrimination between word and pseudoword. The register was carried out for the electrodes of the 10-20 system and the mean amplitudes and latency peaks measured and also the amplitude from 200 to 550 milliseconds. The most significant differences between the ERPs occurred in Cz, with greater negativity for the mean of the amplitude between 425 and 550 milliseconds for pseudowords (N400). The N400 was more precocious in 11 year old. The influence of age was thus evident and also the differences in ERPs between words and pseudowords. PMID- 17119790 TI - Saccadic movements using eye-tracking technology in individuals with autism spectrum disorders: pilot study. AB - OBJECTIVE: To verify differences in the visual scanning strategies between pervasive developmental disorders (PDD) and controls when they are observing social and non-social pictures. METHOD: PDD group (PDDG) comprised by 10 non retarded subjects (age from 4 to 41) and age-matched control group (CG). Nine social pictures with human beings (including two pictures of cat mask), and 3 nonsocial pictures of objects were presented for 5 seconds. Saccadic movements and fixation were recorded with equipment EyeGaze (LC Technologies Inc.). RESULTS: PDDG (mean=292.73, SE=67.62) presented longer duration of saccadic movements for social pictures compared to CG (mean=136.06, SE=14.01) (p=0.04). The CG showed a higher number of fixations in the picture 7 (a women using a cat mask, with the eyes erased) (CG: mean=3.40; PDDG: mean=1.80; p=0.007). CONCLUSION: The results suggest differences in strategies that PDD explore human picture. Moreover, these strategies seem not to be affected by the lack of expected part of the face (the eyes). PMID- 17119791 TI - Attention deficit hyperactivity disorder (ADHD) in adults: social-demographic profile from a university hospital ADHD outpatient unit in Sao Paulo, Brazil. AB - PURPOSE: To describe the social-demographic variables, including interpersonal, academic, and professional performance in adult individuals with the diagnoses of attention deficit hyperactivity disorder (ADHD). There are no reports of this kind in the Brazilian population with ADHD. The ADHD is a common disorder, which can reach up to 3% of the general population. METHOD: Descriptive study of adults with ADHD, according to the DSM-IV criteria (American Psychiatric Association). The sample was selected from a specialized outpatient service in Sao Paulo city. The social-demographic data was obtained by personal interviews. RESULTS: There was a predominance of males in the sample (61.2%) and a high education level (90.2% had at least a high school degree) and 52% of the 102 patients had repeated their class at a least once during their school lives. In addition, 22.5% of the population sample were unemployed at the time of the interview. CONCLUSION: The distribution of the socio-demographic variables in adult ADHD is similar to other ADHD samples reported in other countries, despite the high education level met in our sample. Similarities between child and adult ADHD could also be traced. PMID- 17119792 TI - Vascular parkinsonism: analysis of seven cases. AB - INTRODUCTION: Neuroimaging studies of elderly individuals reveal alterations in the white matter that are incompatible with the patients parkinsonism, mistakenly classified as vascular parkinsonism (VP). METHOD: This study was conducted on a population composed of 20 patients with Parkinsons disease (PD) whose neuroimaging exams revealed vascular alterations in the white matter and seven patients with VP in order to compare diagnostic criteria. RESULTS: Age at disease onset of patients with PD was 55+/-12 years and patients with VP it was 62+/-13 years. Twelve patients with PD and five patients with VP presented arterial hypertension; three patients with VP and two patients with PD presented gait impairment; all patients with VP presented rigidity and bradykinesia, six of them presented resting tremor; 19 patients with PD presented tremor and 19 of them presented rigidity, while 17 presented bradykinesia. When the symptoms and evolution of both diseases were compared, the vascular alterations in the white matter were considered unspecific. CONCLUSION: Since clinical symptoms are unspecific, a differential diagnosis requires neuroimaging, good response to levodopa and clinical evolution. PMID- 17119793 TI - Effects of cervical sympathectomy on vasospasm induced by meningeal haemorrhage in rabbits. AB - This study investigates the role of cervical sympathectomy in the prevention of acute vasospasm induced by meningeal haemorrhage in rabbits. Sixteen adult English Norfolk rabbits were divided into 2 experimental groups: bilateral cervical sympathectomy of the superior sympathetic ganglion (SSSG, n=8), and bilateral SSSG and sympathectomy of the inferior sympathetic ganglion (SISG, n=8). Other 24 animals were used as controls. Basilar artery diameter was evaluated by angiography. SSSG protected the animals against developing cerebral vasospasm; SSSG associated with SISG did not increase this effect. PMID- 17119794 TI - Black, white and shades of grey: SUNCT or short-lasting chronic paroxysmal hemicrania? AB - AIM OF THE STUDY: To report a case of unilateral headache with two possibilities of diagnosis. METHOD: Case report. RESULTS: Patient with unilateral, intense, stabbing periocular headache with conjuntival injection and tearing. Although the duration of attacks was typical of SUNCT, there was complete remission of the pain with indomethacin, suggesting that this was a case of chronic paroxysmal hemicrania with unusually short attack duration. CONCLUSION: Therapeutic trials of indomethacin on younger patients presenting clinical diagnosis of SUNCT could be tried on a more regular basis. PMID- 17119795 TI - Visual and automatic cyclic alternating pattern (CAP) scoring: inter-rater reliability study. AB - The classification of short duration events in the EEG during sleep, as the A stage of the cyclic alternating pattern (CAP) is a tedious and error prone task. The number of events under normal conditions is large (several hundreds), and it is necessary to mark the limits of the events with precision, otherwise the time sensitive classification of the CAP phases (A and B) and specially the scoring of different types of A phases will be compromised. The objective of this study is to verify the feasibility of visual CAP scoring with only one channel of EEG, the evaluation of the inter-scorer agreement in a variety of recordings, and the comparison of the visual scorings with a known automatic scoring system. Sixteen hours of one channel (C4-A1 or C3-A2) of NREM sleep were extracted from eight whole night recordings in European Data Format and presented to the different scorers. The average inter-scorer agreement for all scorers is above 70%, the pair wise inter-scorer agreement found was between 69% up to 77.5%. These values are similar to what has been reported in different type studies. The automatic scoring system has similar performance of the visual scorings. The study also has shown that it is possible to classify the CAP using only one channel of EEG. PMID- 17119796 TI - Andersen syndrome: an association of periodic paralysis, cardiac arrhythmia and dysmorphic abnormalities. AB - Andersen syndrome (AS) is a rare disease characterized by the presence of periodic paralysis (PP), cardiac arrhythmia and dysmorphic abnormalities. We report herein the first Brazilian patient presenting AS who also had obesity, obstructive sleep apnea (OSA) and daytime sleepiness. Clinical and genetic evaluation of six family members demonstrated that four had dysmorphic abnormalities but none had PP or cardiac arrhythmia. Sequencing of KCNJ2 revealed the R218W mutation in the index patient and her 6-year-old daughter, who presented dysmorphic abnormalities (micrognathia, clinodactyly of fourth and fifth fingers, short stature) and OSA. Three relatives had clinodactyly as the only manifestation but the R218W mutation was absent, suggesting that this characteristic may be influenced by another gene. OSA accompanied by dysmorphic features may be related to AS. PMID- 17119797 TI - [Reibergram for C3c intrathecal synthesis evaluation]. AB - INTRODUCTION: Reiber's quotient diagram or reibergram has a growing apply for characterize the intratecal synthesis of proteins. Firstly reibergrama was used for the major classes of immunoglobulins but later it was used to evaluate other proteins based on the theory about molecular flux/cerebrospinal fluid (CSF) flow rate. METHOD: C3c is a degradation product of complement factor C3 with 145 KDa and approaches to IgG molecular characteristics according with Fick's diffusion laws. It was assumed IgG constants and graphic for IgG constants and graphic to evaluate the intrathecal synthesis of C3c. Twenty-seven patients and 27 controls were studied. Serum and CSF C3c and albumin were quantified by immunodiffusion. RESULTS: The patients with the C3c proposed reibergram were evaluated. It has been proved its validity under several CSF blood barrier conditions. CONCLUSION: Reibergram for C3c can be used for the evaluation of the intrathecal synthesis of this protein. PMID- 17119798 TI - sICAM-1 in meningoencephalitis due to Angiostrongylus cantonensis. AB - INTRODUCTION: Angiostrongylus cantonensis meningoencephalitis is an emergent disease in the Americas. METHOD: Twelve children suffering from eosinophilic meningoencephalitis due to this parasite aged between 6-10 years were studied. Cerebrospinal fluid (CSF) and serum samples were taken simultaneously in the first diagnostic puncture at admission. RESULTS: All cases showed typical findings on the routine CSF and serum analysis: increased CSF total protein, increased Q (CSF/serum) albumin accompanied by eosinophilia in CSF. No intrathecal synthesis of immunoglobulins was found. Mean serum and CSF sICAM-1 values were 337.4 and 3.97 ng/mL. Qalbumin and QsICAM-1 mean values were 4.1 and 6.2 respectively. In 50% of the patients an increased brain-derived fraction of sICAM-1 was found. CONCLUSION: It may be suggested that a dynamic of the sICAM-1 brain derived fraction is perhaps associated to the immune response in the evolution of the disease.sICAM-1 may be an agent in negative feedback for eosinophils passage through the blood-CSF barrier into the inflammatory brain response. PMID- 17119799 TI - CSF markers for diagnosis of bacterial meningitis in neurosurgical postoperative patients. AB - OBJECTIVE: To evaluate the diagnostic usefulness of cerebral spinal fluid (CSF) cellularity, protein, neutrophils, glucose and lactate for detection of postoperative bacterial meningitis. METHOD: This prospective study was conducted in 28 postoperative neurosurgical patients from 2002 to 2005 at University of Sao Paulo. The CSF markers were plotted in a receiver operating characteristic (ROC) curve to evaluate their accuracy. RESULTS: Based on the area under ROC curve CSF glucose, cellularity, and lactate were considered good tests. Polymorphonuclear and protein did not achieve enough accuracy to be used clinically. CONCLUSION: The CSF glucose, lactate, and cellularity can be used for the diagnosis of bacterial meningitis. Moreover, it can be helpful to differentiate bacterial from aseptic meningitis. PMID- 17119800 TI - Wrist immobilization after carpal tunnel release: a prospective study. AB - This prospective study evaluates the possible advantages of wrist immobilization after open carpal tunnel release comparing the results of two weeks immobilization and no immobilization. Fifty two patients with idiopathic carpal tunnel syndrome were randomly selected in two groups after open carpal tunnel release. In one group (A, n=26) the patients wore a neutral-position wrist splint continuously for two weeks. In the other group (B, n=26) no wrist immobilization was used. Clinical assessment was done pre-operatively and at 2 weeks follow-up and included the two-point discrimination test at the second finger and two questionnaires as an outcome measurement of symptoms severity and intensity. All the patients presented improvement in the postoperative evaluations in the three analyzed parameters. There was no significant difference between the two groups for any of the outcome measurements at the final follow-up. We conclude that wrist immobilization in the immediate post-operative period have no advantages when compared with no immobilization in the end result of carpal tunnel release. PMID- 17119801 TI - Peripheral neuropathy in patients with hepatitis virus C infection in the Amazon region. AB - INTRODUCTION: Hepatitis virus C (HCV) infection is considered a health problem in the State of Acre localized in the Brazilian Amazon which has a prevalence rate of 5.9%. Peripheral neuropathy is a common extra-hepatic manifestation in patients with HCV. OBJECTIVE: To determine the prevalence of peripheral neuropathies using clinical and neurophysiological parameters. METHOD: A cross sectional study was performed in patients assisted by a specialized center of infectious diseases in the State of Acre. All patients completed a clinicoepidemiological questionnaire, physical examination and nerve conduction studies (NCS). RESULTS: We studied 78 patients with mean age 45.5 years (range from 10 to 76 years), two thirds were male, 51% had at least 8 years of formal education and 96% lived in the capital city of Acre State. Roughly 34% of patients complained about paresthesias mainly in upper limbs. The NCS diagnosed multiple mononeuropathy in 11 (14.1%; IC95% 7.6-23.2) patients and carpal tunnel syndrome in 4 (5.1%) patients. CONCLUSION: Subclinical involvement of peripheral nerves seems common in patients with HCV, with multiple mononeuropathy the main manifestation of nerve injury in this region as suggested by electrophysiological studies. PMID- 17119802 TI - Isolated velopalatine paralysis associated with parvovirus B19 infection. AB - A case of isolated velopalatine paralysis in an 8-year-old boy is presented. The symptoms were sudden-onset of nasal speech, regurgitation of liquids into the nose and dysphagia. Brain MRI and cerebrospinal fluid examination were normal. Infectious serologies disclosed an antibody arrangement towards parvovirus B19 that was typical of recent infection. In the absence of other positive data, the possibility of a correlation between the tenth nerve palsy and parvovirus infection is discussed. PMID- 17119803 TI - Episodes of Guillain-Barre syndrome associated with the acute phase of HIV-1 infection and with recurrence of viremia. AB - We report a severe case of Guillain-Barre syndrome (GBS) characterized by flaccid areflexive tetraplegia and signs of autonomic instability related to acute HIV-1 infection, and the occurrence of relapse episodes coinciding with the detection of HIV-1 RNA in blood during the phase of irregular treatment with antiretroviral agents. The patient has been asymptomatic for 3 years and has an HIV-1 load below the limit of detection. The recurrence of GBS in this case may be related to alterations of the immunologic response caused by disequilibrium in the host-HIV relationship due to the increase in HIV-1 viremia. PMID- 17119804 TI - Acute axonal polyneuropathy with predominant proximal involvement: an uncommon neurological complication of bariatric surgery. AB - Bariatric surgery is frequently indicated in the treatment of morbid obesity. Previously unreported complications have been associated to this surgery; among them, neurological complications have gained attention. We report the case of a 25-year-old man submitted to gastric surgery for treatment of morbid obesity who developed, two months after surgery, acute proximal weakness in lower limbs. The electroneuromyography revealed axonal peripheral polyneuropathy with predominant proximal involvement. After treatment with immunoglobulin and vitamin supplementation, rapid clinical and neurophysiologic recovery was observed. We describe the clinical and electroneuromyographic features of this case, stressing the difficulty of initial diagnosis, particularly in the differential diagnosis with Guillain-Barre syndrome. We discuss the importance of nutritional follow-up and the eventual indication of routine vitamin supplementation in these patients. PMID- 17119805 TI - Ganglioglioma: comparison with other low-grade brain tumors. AB - METHOD: Forty-two patients with low-grade brain tumor and refractory epilepsy were studied. The mean age was 22.3 years. They were divided into two groups: Group A, patients with ganglioglioma (n=19) and group B, patients with other low grade tumors (n=23) (14 astrocytoma, 6 oligodendroglioma, 2 dysembryoplastic neuroepithelial tumor, and 1 xanthoastrocytoma). RESULTS: Age at seizures onset was 7 years or less in 73% of the patients in group A and in 30.4% of the patients in group B (p=0.045). Complex partial occurred frequently in group A and B (94.7% versus 82%, respectively). Seizures frequency was higher in group B (p=0.002). Computerized tomography (CT) was normal in 36.8% of group A patients and abnormal in all group B patients. Magnetic resonance imaging (MRI) was abnormal in all patients. Surgical removal was complete in 89.5% of the patients in group A and in 78.2% of the patients in group B. CONCLUSION: The association of refractory epilepsy and complex partial seizures, at a relatively low frequency, in young patients potentially normal CT and a MRI hypointense temporal lobe lesion in T1-weighed slices were habitual image findings in ganglioglioma, rather than other low-grade tumor. PMID- 17119806 TI - Abnormalities in auditory evoked potentials of 75 patients with Arnold-Chiari malformations types I and II. AB - OBJECTIVE: To evaluate the frequency and degree of severity of abnormalities in the auditory pathways in patients with Chiari malformations type I and II. METHOD: This is a series-of-case descriptive study in which the possible presence of auditory pathways abnormalities in 75 patients (48 children and 27 adults) with Chiari malformation types I and II were analyzed by means of auditory evoked potentials evaluation. The analysis was based on the determination of intervals among potentials peak values, absolute latency and amplitude ratio among potentials V and I. RESULTS: Among the 75 patients studied, 27 (36%) disclosed Arnold-Chiari malformations type I and 48 (64%) showed Arnold-Chiari malformations type II. Fifty-three (71%) of these patients showed some degree of auditory evoked potential abnormalities. Tests were normal in the remaining 22 (29%) patients. CONCLUSION: Auditory evoked potentials testing can be considered a valuable instrument for diagnosis and evaluation of brain stem functional abnormalities in patients with Arnold-Chiari malformations type I and II. The determination of the presence and degree of severity of these abnormalities can be contributory to the prevention of further handicaps in these patients either through physical therapy or by means of precocious corrective surgical intervention. PMID- 17119807 TI - [Transspheinoidal transpalatal meningoencephalocele]. AB - Transspheinoidal meningoencephalocele is a rare presentation of dysraphism of the neural tube. It is characterized by the herniation of the neural tissue through a bony defect in the sphenoid bone. The clinical presentation is variable. It may be assymptomatic or it may include an upper airway obstruction, rhinorrhea, meningitis, hypothalamic dysfunction and optic anomalies. The surgical treatment is controversial. We describe the case of a 7-year-old boy who presented a pulsate structure filling the palate, palate digenesis and hypertelorism. The diagnosis of transsphenoidal transpalatal meningoencephalocele was confirmed by a computerized tomography and a magnetic resonance imaging. The child was operated on by the transpalatal/transspheinoidal approach with a good result. PMID- 17119808 TI - [Neuroimaging of the dopamine transporter in Parkinsons disease: first study using [99mTc]-TRODAT-1 and SPECT in Brazil]. AB - BACKGROUND: Dopamine transporter (DAT) neuroimaging radiotracers were developed to estimate dopamine neuronal loss in vivo in Parkinsons disease (PD). OBJECTIVE: To evaluate DAT density in vivo using [99mTc]-TRODAT-1 and single photon computerized tomography (SPECT) in a population of Brazilian PD. METHOD: Fifteen PD patients and 15 matched healthy controls scanned with [99mTc]-TRODAT-1 (INER Taiwan) and SPECT. Estimates of striatum DAT density were calculated using binding potential (BP). Patients were assessed with PD scales. RESULTS: PD patients had significantly lower striatal DAT-BP (mean+/-SD) (0.38+/-0.12) compared to controls (BP=0.84+/-0.16; p<0.01). A 100% sensitivity and 100% specificity was obtained to discriminate PD cases from controls. Negative correlations between striatal DAT-BP and PD severity (rho=-0.7, p<0.001) and motor scales (rho=-0.80, p<0.001) were found. CONCLUSION: [99mTc]TRODAT-1 SPECTs scanning was able to discriminate PD patients from controls. The technique is a powerful instrument to measure DAT density that can be used in clinical and research settings in Brazil. PMID- 17119809 TI - [Clinical and epidemiologic characteristics of 20 patients with multiple sclerosis in Cuiaba-Mato Grosso, Brazil]. AB - Multiple sclerosis is an acquired immune-mediated disease of the central nervous system that show a widely variation of clinical aspects. In Brazil, demographic and ethnical aspects show these variations as has been demonstrated by many Brazilian papers. This retrospective study evaluated epidemiological and clinical characteristics of 20 multiple sclerosis patients followed in Cuiaba, situated on Brazil middle west region. The patients were classified followed at Neurology Ambulatory of CRIDAC from 1998 to 2005. The data show a 3:1 female/male ratio and that 20% of patients were from black origin. EDSS medium range was 3.9; 55% of patients were born in Mato Grosso State. The most common symptoms were pyramidal (90%), cerebellar (60%) and visual (25%). All patients were on immunomodulatory or immunosupressive treatment. PMID- 17119810 TI - [Morphological characteristics from the insula's lobe in patients with medial temporal lobe epilepsy]. AB - The temporal medial sclerosis (TMS) is characterized by hippocampal sclerosis in temporal and by distinguished grades of injury near to other neurological structures such as: amygdaloid nucleus, parahippocampal girus and entorhinal region. The study analyzed 40 patients with TMS and 40 people from the control cluster. All the cases were appreciated by one method for measurement of insula's cortex (E-Film) and another method to calculate the insula's volume (Neuroline). There is no variation statistical between the insula's volume and insula's measurement for the two clusters. This paper didn't show the insula's morphological variation when these two groups were compared. PMID- 17119811 TI - [Severe behavioral changes in a patient with Fahr's disease]. AB - We report on a case of a 40 year-old man with Fahrs disease, defined by idiopathic bilateral basal ganglia calcification, who developed depressive disorder, motor and phonic tics, stereotyped behaviors such as punding and personality changes with significant social and familiar implications. We discuss about the psychopathology of Fahrs disease and the relevance of the basal ganglia in the determination of humans behavior. PMID- 17119812 TI - Endoscopic assisted microneurosurgery for gasserian portion of trigeminal neuroma: two cases. AB - We report two cases of trigeminal neuroma that were operated on by the neurosurgery team at Felicio Rocho Hospital, Belo Horizonte, Minas Gerais State, Brazil. Endoscopic assisted microsurgery was the technique used to approach the gasserian region tumor with good results. PMID- 17119813 TI - Primary Ewings sarcoma of the spine: case report. AB - Primary malignant sarcomas of the spine are extremely rare. Because of biological heterogeneity, these tumors have variable sensitivity to radiation and chemotherapy. Adequate local control through complete tumor removal is an important therapeutic goal. However, aggressive resection of tumors in the spinal column must be coupled with restoration of spinal column stability and minimization of neural deficits. The balance of these factors makes treatment of primary sarcomas of the spine challenging, and dictates an individual approach to treatment. We report on a 18 years old man with primary Ewing's sarcoma of the nonsacral spine. The clinical picture and imaging characteristics were analyzed as well as the management modalities and outcome. PMID- 17119814 TI - Cerebral ischemia as initial neurological manifestation of atrial myxoma: case report. AB - Cerebral infarctions of cardiac etiology are observed in around 20% of patients with ischemic stroke. Cerebral ischemia is the first clinical manifestation in 1/3 of cases of atrial myxomas. Although almost half of patients with atrial myxoma show changes at neurological exam, non-hemorrhagic cerebral infarction is seen in computed tomography in practically all cases. We present the case of a 40 year-old woman whose first clinical manifestation of atrial myxoma was an ischemic stroke. We point out to the possibility of silent cerebral infarction in atrial myxoma patients. PMID- 17119815 TI - Symptomatic non-atherosclerotic bilateral extracranial vertebral artery occlusion treated with extracranial to intracranial bypass: case report. AB - Posterior fossa ischemia is not a very frequent situation. It is responsible for about 25% of all ischemic strokes, and the vast majority of the cases are related to atherosclerotic stenosis of the vertebral and/or basilar arteries. Acute ischemia can also occur in the setting of vertebral artery dissection, traumatic or spontaneous. Recently, blunt trauma has been increasingly recognized as a cause for craniocervical artery injury. The management options for both traumatic and atherosclerotic lesions of the posterior fossa are still under debate. We present a case of a delayed onset of hemodynamic ischemic symptoms due to bilateral vertebral artery occlusion probably related to remote trauma to the head and neck in a 55-year-old-man treated successfully with extracranial to intracranial bypass. PMID- 17119816 TI - Basilar impression associated with impacted cisterna magna, spastic paraparesis and distress of balance: case report. AB - We report on a 48 years-old man with basilar impression without syringohydromyelia, in which the cisterna magna was impacted by the cerebellar tonsils. Six months after posterior fossa decompression there was the disappearance of nuchal rigidity, vertigo, spastic paraparesis and improvement of balance. Nevertheless hyperreflexia and diminished pallesthesia of the lower limbs persisted. PMID- 17119817 TI - Impacted cisterna magna without syringomyelia associated with spastic paraparesis: case report. AB - We report on a 49 year old man with impacted cisterna magna without the presence of syringohydromyelie (SM). The clinical picture was characterized by spastic paraparesis. Magnetic resonance imaging depicted a cisterna magna filled by the cerebellar tonsils. Six months after osteodural-neural decompression of the posterior fossa there was resolution of neurological symptoms and signs with the exception of hyperactive patellar and Achilles reflexes. PMID- 17119818 TI - Intracranial arterial aneurysms in childhood: case report. AB - A case of an intracranial arterial aneurysm at internal carotid bifurcation in a 10-year-old girl is described with the special features of cerebral aneurysm which occur in children, comparing with the adults. We alert for the necessity of carefully operative technique in order to avoid damage and intraoperative rupture of the aneurysm due to the very thin vessel wall that this population can develop. Our recommendation is early surgery in these patients. PMID- 17119819 TI - [Spontaneous occlusion of previously cerebral embolized residual arteriovenous malformation: report of two cases]. AB - OBJECTIVE: To report two cases of spontaneous occlusion of residual malformations that had previously been embolized and to analyze their probable causes. METHOD: Retrospective review of patients with embolized arteriovenous malformations (AVMs) between January 1999 and August 2005 in our institution and analysis of two cases of occlusion in residual AVMs. RESULTS: Two cases of AVMs that received endovascular treatment and had post-embolization residual nidus malformation, which in later angiographic controls showed a complete disappearance of the lesions without other treatment. CONCLUSION: The spontaneous occlusion of residual arteriovenous malformations is an infrequent occurrence. Several related mechanisms are discussed. PMID- 17119820 TI - [Paracoccidioidomycosis in cerebral hemisphere and brainstem: case report]. AB - We report on a 36 years-old man that had been at the Amazon forest four years before. Six months before the admission he had developed a progressive quadriparesis, gait ataxia, dysphagia, dysarthria, difficulty in breathing and hiccup. The gadolinium-enhanced T1-weighted MRI showed a lesion into the right parietoccipital area and another into the medulla, that was the largest. There was any evidence of tuberculosis or AIDS. The patient was submitted to microsurgical approach to the medulla. Pathological examination revealed paracoccidioidomycosis. Treatment with anphotericin B till 2100 mg was administered followed by sulfamethoxazole-trimetoprim for three months plus physical therapy. The patient went back to his activities six months after the end of the treatment. Comments are presented about the participation of the immunological system and of the cytokines (interleukines). PMID- 17119821 TI - [Reliability of neurological assessment scales in patients with stroke]. AB - Scales for the assessment of neurological impairment are employed in clinical trials, outcome evaluation and in therapeutic decisions. We evaluated the reliability of the Portuguese version of the Rankin Scale (RS), Barthel Index (BI) and the NIHSS (National Institute of Health Stroke Scale) in 51 stroke patients, estimating the agreement of the results for examiners. The interrater reliability was measured using the Kappa Coefficient and Intraclass Correlation Coefficient. The RS showed moderate, substantial and excellent coefficients of agreement. The BI showed from substantial to excellent coefficients of agreement as a whole and also for its constituents. This scale was stratified in prognostic and functional groups, both presenting coefficients of agreement from substantial to excellent. The NIHSS presented excellent agreement on its total and substantial agreements on its individual items. The NIHSS was also stratified in groups of points, which presented moderate or excellent coefficient, and in groups of congruous items, wich showed a moderate to excellent agreement. These results suggest that the Portuguese version of the RS, BI and the NIHSS present adequate reliability. PMID- 17119822 TI - [Basal ganglia calcification on computed tomography]. PMID- 17119824 TI - Species composition and structure of Thysanoptera communities in different microhabitats at the Parque Estadual de Itapua, Viamao, RS. AB - Although thrips are known as inhabitants of flowers, they are also abundant and diverse in other microhabitats. There is an information gap concerning them, especially related to the native fauna in southern Brazil. The structure and composition of the thysanopteran community in different microhabitats was studied at the "Parque Estadual de Itapua" (30 degrees 22' S 51 degrees 02' W), RS, southern Brazil. Between June 1999 and May 2001, branches (n = 1,274), flowers (n = 774), grass tussocks (n = 596) and leaf litter (n = 603) were sampled systematically in 20 points of four trails (T1 - Pedreira beach, T2 - Araca beach, T3 - Lagoinha, and T4 - Grota hill). We found 2,197 adult thrips determined in 73 species in 41 genera, of which 37 could be nominated. Four families are represented, Thripidae, Phlaeothripidae, Heterothripidae and Merothripidae, with the first the most abundant (N = 1,599) and with the highest species richness (S = 32). The highest thrips abundance occurred in flowers N = 1,224 and the highest number of exclusive species occurred in the leaf litter (27). Frankliniella rodeos Moulton, 1933, Frankliniella gemina Bagnall, 1919 and Smicrothrips particula Hood, 1952 comprise 49.4% of the total sampled. Regarding T2, we obtained the highest abundance (N = 935) and highest species richness (S = 43). The composition of the faunas in each kind of environment proved very particular. PMID- 17119825 TI - Changes in biomass, chemical composition and nutritive value of Spartina alterniflora due to organic pollution in the Itanhaem River Basin (SP, Brazil). AB - We compared the values of the biomass, chemical composition and nutritive value of the emergent aquatic macrophyte S. alterniflora in a river affected by the discharge of domestic sewage (Guau River) and in an unpolluted river (Itanhaem River). S. alterniflora, water and sediment samples were obtained in the two rivers in November, 2001. The Guau River presented the highest levels of Total-N and Total-P in the water (415 and 674 microg.L(-1), respectively) and in the sediment (0.25 e 0.20% of the Dry Mass, respectively), when compared to the water (TN = 105 microg.L(-1); TP = 20 microg.L(-1)) and the sediment (NT = 0.12% DM; PT = 0.05% DM) of the Itanhaem River. Aerial (316 g DM.m(-2)) and subterraneous (425 g DM.m(-2)) biomass of S. alterniflora were significantly higher in the Guau River than in the Itanhaem River (146 and 115 g DM.m(-2), respectively). In addition, the values of TN, protein, TP, lipids and soluble carbohydrates were significantly higher in S. alterniflora biomass from the Guau River. On the other hand, the values of the polyphenols and the cell wall fraction were significantly higher in the biomass of S. alterniflora from the Itanhaem River. We concluded that domestic sewage discharge in water bodies may increase the biomass and change the chemical composition of S. alterniflora. The high N and P availability in the water of the Guau River is probably the cause of the higher biomass, TN, TP, protein, lipids and soluble carbohydrates measured in S. alterniflora in this river. PMID- 17119826 TI - A medium-spatial scale distribution pattern of Pseudoscorpionida (Arachnida) in a gradient of topography (altitude and inclination), soil factors, and litter in a central Amazonia forest reserve, Brazil. AB - In Amazonia, nothing is known about the distribution of the invertebrates on a medium-spatial scale pattern. In a trail system of 64 km2 at Ducke Reserve, we sampled 72 transects using the hand-sorting method and Berlese-Tullgren extraction. The reserve possesses ecosystems of "terra-firme" forest and the trail system represents a gradient of topographic soil factors and vegetation, avoiding categorizations. Considering the abundance and diversity of Pseudoscorpionida, we investigated the relation of the community to environmental factors tested (topography, clay percentage, litter, and soil pH), to the two major drainage basins of the reserve, and if these invertebrates can be used as biological indicators to monitor changes. We registered two species for the first time in the reserve, increasing the known diversity to 17 species. The lack of correlation with the predictor variables and the large home range, indicate that pseudoscorpions are not good biological indicators in the reserve. As the eastern and western watersheds are not separate management units for the community, our results show that they are generalist predators. In spite of our results and lack of knowledge concerning their biology, life history and taxonomy, pseudoscorpions are cosmopolitan and easy to find and measure. Compared with previous studies in the reserve, they have a consistent pattern of abundance and diversity throughout the years showing the stability of the community which can be checked mainly by comparison with environmental changes that would occur in the reserve. An investigation on a medium-spatial scale pattern and over a long-term period including other habitats, and also other predictor variables, like humidity, the structure of the vegetation and canopy closure, will be necessary to reinforce the observed tendencies. PMID- 17119827 TI - Diet and foraging of the endemic lizard Cnemidophorus littoralis (Squamata, Teiidae) in the restinga de Jurubatiba, Macae, RJ. AB - We investigated the diet and foraging of the endemic teiid lizard Cnemidophorus littoralis in a restinga habitat in Jurubatiba, Macae - RJ. The stomach contents were removed, analyzed and identified to the Order level. There was no relationship between C. littoralis morphological variables and number, length or volume of preys. Termites (48.7%) and larvae (35.5%) were the most important prey items which occurred in the examined lizards' stomachs. The diet did not differ between males and females. Cnemidophorus littoralis is an active forager and predominantly consumes relatively sedentary prey or prey that is aggregated in the environment. We also found an intact and undigested hatchling of the crepuscular/nocturnal gekkonid lizard Hemidactylus mabouia in the stomach of an adult male of C. littoralis, which indicates that C. littoralis is a potential source of mortality for individuals of H. mabouia in the restinga de Jurubatiba. PMID- 17119828 TI - Ecological-evolutionary relationships in Passiflora alata from Rio Grande do Sul, Brazil. AB - The geographical distribution, ecological characteristics, flowering and fruiting times, and pollinating agents of Passiflora alata are considered and related to molecular genetic data gathered simultaneously. The first report on this species in Rio Grande do Sul was made in 1934, only in cultivated gardens. Approximately 20 years later, however, the species was already classified as efferata (wild) in Porto Alegre's suburbs. The data presented here, together with the DNA investigations, indicate that P. alata is actively colonizing previously unoccupied areas of this region. PMID- 17119829 TI - Income evaluation of small - scale fishers in two Brazilian urban reservoirs: Represa Billings (SP) and Lago Paranoa (DF). AB - The income of small-scale fisheries of two urban reservoirs in Brazil: Represa Billings (127 km2) located in the metropolitan region of Sao Paulo, the capital city of the state of Sao Paulo and Lago Paranoa (38 km2) located in Brasilia (DF), the capital city of Brazil were calculated and compared. Both fisheries are mainly based on the alien Nile tilapia Oreochromis niloticus (more than 80% of the total catch). Although these reservoirs are nearly 900 km apart, their native fish fauna belong to the Upper Parana Province. The Represa Billings fishers have, on average, a daily profit of 15.8 R dollars (8.4 U dollars mostly employing gill nets) and Lago Paranoa fishers 46.6 R dollars (24.9 U dollars, mostly employing cast nets), which is a statistically significant difference (P < 0.001). The profit of the fisheries is explained by the factor "reservoir" and covariate "days of fishing". Due to the increasing violence in the metropolitan region of Sao Paulo, the Represa Billings fishery is vanishing. PMID- 17119830 TI - Protected cerrado fragments grow up and lose even metapopulational birds in central Sao Paulo, Brazil. AB - Moderately dense woodland (cerradao) grew in two isolated patches of bushy savanna (cerrado) in central Sao Paulo over 23 years of bird censuses. Various uncommon birds were lost and some forest species were permanently gained. Fall and winter fruits attract long and short-distance migrants. Woodpeckers and some birds that nest in their holes seem to disappear during tree growth. Some birds in weedy areas nearby disappeared when the pastures replaced these areas, however sugar cane reduced the numbers of birds even more up to the point when some areas became pastures once more. Even travel-prone species disappear with vegetation growth in cerrado protected fragments, and therefore "metapopulations" may not survive over time, only in space. PMID- 17119831 TI - Sticholonche zanclea (Protozoa, Actinopoda) in fecal pellets of copepods and Euphausia sp. in Brazilian coastal waters. AB - Fecal pellets produced by mesozooplanktonic copepods (Centropages velificatus and Paracalanus parvus) and macrozooplanktonic Euphausiacea (Euphausia sp.) were examined using scanning electron microscopy. Fragments of the protozoan Sticholonche zanclea were found in both copepod and in Euphausia sp. fecal pellets, even when the abundance of the protozoan in the water was low. The results suggest that S. zanclea is an important food resource for different trophic levels, including meso- and macrozooplankton, in Brazilian coastal waters. PMID- 17119832 TI - Study of a community of flies at different altitudes in the Serra of Caldas Novas Park, Goias, Brazil. PMID- 17119833 TI - Phenotypic plasticity in Passiflora suberosa L.(Passifloraceae): induction and reversion of two morphs by variation in light intensity. AB - Leaf morphology may vary considerably even within a branch of Passiflora suberosa plants. Leaves are of a typical green type in shaded areas, but in open fields turn into violet, and apparently have greater thickness and trichome density. The proximate causes and the adaptive meaning, if any, for the existence of the violet morph are still unknown. By cultivating P. suberosa clones under two light regimes (total and partial exposure to sunlight), we consecutively induced (first year) and then reversed (second year) the appearance of the violet morph. We evaluated the corresponding changes in morpho-anatomic and chemical leaf characteristics. Plants that were grown under partial sunlight had a greater size and did not alter their green color, but those grown under total sunlight changed into violet, were smaller in size and their leaves were tougher, thicker, and had a greater number of trichomes. The violet morph had increased anthocyanins and phenolic derivatives. It also showed cellular hypertrophy, a greater number of cell layers in the mesophyll, and a lignified pericycle. Since these morphs are interchangeable by changing light conditions, we inferred that they are not determined by genotypic diversity, but are mainly a result of a physiological response to light stress, and thus part of P. suberosa phenotypic plasticity. PMID- 17119834 TI - Behavioural and chemical studies of discrimination processes in the leaf-cutting ant Acromyrmex laticeps nigrosetosus (Forel, 1908). AB - Leaf-cutting ants live in symbiosis with a basidiomycete fungus that is exploited as a source of nutrients for ant larvae. Tests of brood transport revealed that Acromyrmex laticeps nigrosetosus workers did not discriminate a concolonial brood from an alien brood. The same result was observed with tests of fungus transport. Adult workers showed no aggressive behaviour to workers from other alien colonies (non-nestmates). There was no qualitative variation in the chemical profiles of larvae, pupae and adult workers from the different colonies. However, quantitative differences were observed between the different colonies. Hypotheses about the lack of intraspecific aggression in this subspecies of ants are discussed. PMID- 17119835 TI - Phylogenetic autocorrelation and evolutionary interpretation of the higher-taxon approach for biodiversity analyses. AB - Although in most recent broad-scale analyses, diversity is measured by counting the number of species in a given area or spatial unity (species richness), a 'top down' approach has been used sometimes, counting higher-taxon (genera, family) instead of species with some advantages. However, this higher-taxon approach is quite empirical and the cut-off level is usually arbitrarily defined. In this work, we show that the higher-taxon approach could be theoretically linked with models of phenotypic diversification by means of phylogenetic autocorrelation analysis in such a way that the taxonomic (or phylogenetic) rank to be used could not be necessarily arbitrary. This rank expresses past time in which taxa became independent for a given phenotypic trait or for the evolution of average phenotypes across different traits. We illustrated the approach by evaluating phylogenetic patches for 23 morphological, ecological and behavioural characters in New World terrestrial Carnivora. The higher-taxon counts at 18.8 mya (S(L)) defined by phylogenetic correlograms are highly correlated with species richness (r = 0.899; P < 0.001 with ca. 13 degrees of freedom by taking spatial autocorrelation into account). However, S(L) in North America is usually larger than in South America. Thus, although there are more species in South and Central America, the fast recent diversification that occurred in this region generated species that are "redundant" in relation to lineages that were present at 18.8 my. BP. Therefore, the number of lineages can be comparatively used as a measure of evolutionary diversity under a given model of phenotypic divergence among lower taxonomic units. PMID- 17119836 TI - Seed shadow, seedling recruitment, and spatial distribution of Buchenavia capitata (Combretaceae) in a fragment of the Brazilian Atlantic forest. AB - Here we describe the seed shadow, seedling recruitment, ontogenetic structure and spatial distribution of Buchenavia capitata (an emergent canopy tree) in a 380-ha fragment of the Atlantic forest in northeast Brazil. In particular, we examine seed distribution around 10 parental trees and both seedling recruitment and mortality, during an 18 month period beneath and around parental trees. Moreover, we describe: (1) B. capitata occurrence within treefall gaps; (2) population structure in terms of ontogenetic stages for the whole site; and (3) spatial distribution of adults within an area of 51 hectares. 99% of seeds were found beneath parent crowns (n = 4,236) and seed density reached 14.6 +/- 29.9 seeds/m2 (0-140 seeds/m2). 49% of all seeds germinated but seedling mortality reached 100% after an 18 month period. In addition, saplings of B. capitata were not found in forest understory and within 30 treefall gaps (94-2,350 m2). The adults showed an average DBH of 69.3 +/- 22.1 cm, were 19.2 +/- 2.9 m tall and presented a clumped spatial distribution. B. capitata matched some of the features presented by shade intolerant trees or large-gap specialists, and we hypothesize that low rates or even lack of long distance seed dispersal events may be reducing the probability of B. capitata seeds reaching suitable habitats for successful seedling recruitment and growth. Because of that (1) seedlings face high levels of early mortality; (2) there is no sapling recruitment at the study site; and (3) local population faces senility and it is threatened by local extinction. PMID- 17119837 TI - Spiders of soybean crops in Santa Fe province, Argentina: influence of surrounding spontaneous vegetation on lot colonization. AB - Trials during two consecutive soybean cycles were performed in central Santa Fe in order to determine the main spider families present in the crop and to determine the influence of spontaneous margin flora on colonization towards the lot. Samplings were done by sweeping net and pitfall traps. It was concluded that:1. Oxyopidae was the most frequent family in the herbaceous layer of both the margins and the soybean crop, and Lycosidae in the lower layer; 2. Margin strips in a soybean lot contribute to the colonization of the crop by spiders of aerial habits and also promote re-colonization following pesticide applications, since they act as shelters. The influence on spiders of terrestrial habits was somewhat lower; 3. The distribution of the populations of spiders of terrestrial habits was homogeneous in a soybean crop seeded directly and these predators had a greater capacity to control pests at all points of the lot. PMID- 17119838 TI - Reproductive performance of the mite Brevipalpus phoenicis (Geijskes, 1939) on citrus and coffee, using life table parameters. AB - The flat-mite Brevipalpus phoenicis (Geijskes, 1939) (Acari: Tenuipalpidae) is considered important in citrus (Citrus spp.) and coffee plants (Coffea spp.) in Brazil, and is known as the leprosis and coffee ring spot mite, as being a vector of the Citrus Leprosis Rhabdovirus - CitLV and Coffee Ring Spot Virus - CoRSV. The objective of this work is to find out about the reproductive success of B. phoenicis on citric fruits and coffee leaves by fertility life table parameters and its biology. The experiments were carried out in laboratory conditions at 25 +/- 2 degrees C, 70 +/- 10% of relative humidity and 14 h of photophase. The lengths of embryonic and post-embryonic periods were different due to the host where the mite was reared. B. phoenicis showed better development and higher survival and fecundity in citric fruits than coffee leaves. The intrinsic rate of the population increase (r(m)) was 0.128 and 0.090 - females/female/day on citric fruits and coffee leaves, respectively. The citric fruits were more appropriate for the development of B. phoenicis than coffee leaves. PMID- 17119839 TI - Notes on the biology of Trypoxylon (Trypargilum) opacum Brethes (Hymenoptera; Crabronidae) in Southern Brazil. AB - The present study investigated the abundance, seasonality and various life history traits of Trypoxylon (Trypargilum) opacum. Using trap-nests, 320 nests of T. opacum were collected in the Parque Municipal das Araucarias in Southern Brazil (25 degrees 23' 36" S and 51 degrees 27' 19" W) over a 3 year period. Nesting was more frequent during the warm season. Nests consisted of a linear series of 1 to 8 brood cells separated by mud partitions, usually followed by an empty vestibular cell and final-closure mud plug. Brood cells were most commonly provisioned with spiders of the family Araneidae. Sex-ratio was strongly female biased, 3.4:1 females:males. Natural enemies attacking nests T. opacum included chrysidids, ichneumonids, sarcophagids, bombyliids and ants. PMID- 17119840 TI - Nesting biology of Trypoxylon (Trypargilum) lactitarse Saussure (Hymenoptera, Crabronidae) in trap-nests in Southern Brazil. AB - This study was carried in the Parque Municipal das Araucarias in the municipality of Guarapuava, state of Parana, Southern Brazil. Three hundred and sixty five nests of T. lactitarse were obtained using trap-nests of 0.7, 1.0, and 1.3 cm in diameter. All of them had similar architecture, regardless of the diameter of the trap-nest. Completed nests consisted of a linear series of brood cells whose average number per nest was of 3.3, 4.0 and 3.6 for the nests with 0.7 cm, 1.0 cm and 1.3 cm in diameter, respectively. They were constructed more often during the summer. T. lactitarse had two types of life cycles: direct development (without diapause), and delayed development (with diapause during winter). Natural enemies included Chrysididae, Sarcophagidae, Dolichopodidae and Ichneumonidae. Out of 1,353 identified spider prey, 1,313 belonged to the Araneidae family. PMID- 17119841 TI - Parasitic community of Fransciscodoras marmoratus (Reinhardt, 1874) (Pisces: Siluriformes, Doradidae) from the upper Sao Francisco River, Brazil. AB - One hundred and thirteen specimens of Franciscodoras marmoratus (Reinhardt, 1874) were collected in the upper Sao Francisco River (18 degrees 12' 32" S, 45 degrees 15' 41" W, state of Minas Gerais) between September, 1999 and January, 2004 to investigate their parasite fauna. From this total, 45 (39.8%) were afflicted by at least one parasite species. The parasitic richness consisted of six species represented by Hirudinea (n = 20), Monogenoidea (n = 25), Eucestoda (n = 55), Nematoda (n = 1, n = 2) and Acanthocephala (n = 41) found in the dry and wet periods making a total of 144 specimens. Proteocephalus renaudi Chambrier & Vaucher, 1994 was the only species with prevalence higher than 10% and a typical aggregate distribution pattern. The prevalence, intensity and abundance of P. renaudi were not influenced by the total length or sex of the hosts or by the collection periods. The relative condition factor indicated that the health of the P. renaudi hosts was not significantly affected in relation to fish not infected by parasites. The fish stocked in tanks before necropsy were opportunistically infested by Lernaea cyprinacea Yashuv, 1959. The various parasites found indicate that F. marmoratus is omnivorous and a potential definitive host. The parasite species, except for Acanthocephala, have expanded their known geographic distribution to the Sao Francisco River Basin. The parasite community was considered isolationist because of the low endoparasite diversity, infrapopulations with low intensity, lack of evidence of parasite interactions and sparse signs of parasite aggression against their hosts. PMID- 17119842 TI - Phenotypic aspects of oral strains of Candida albicans in children with Down's syndrome. AB - The aim of this article is to characterize the biological aspects of oral strains of C. albicans in children with Down's syndrome. These yeasts were analyzed as to their macromorphological and enzymatic aspects and were tested as to their in vitro susceptibility to antifungal drugs using broth microdilution to determine the minimum inhibitory concentration (MIC). The morphotyping revealed that all oral C. albicans isolates from children with Down's syndrome promoted the formation of fringes regardless of size, while the control group presented smaller fringes. All oral C. albicans strains produced proteinase, but those with phospholipolytic activity showed greater enzyme capacity in the test group. In vitro susceptibility showed that all oral C. albicans isolates were sensitive to the drugs used. PMID- 17119843 TI - Thermal biology of Liolaemus occipitalis (Squamata, Tropiduridae) in the coastal sand dunes of Rio Grande do Sul, Brazil. AB - The thermal biology of the small sand lizard, Liolaemus occipitalis, was studied in the coastal sand dunes at Quintao Beach (Palmares do Sul, Rio Grande do Sul, Brazil; 30 degrees 24' S and 50 degrees 17' W), between September, 1998 and August, 1999. Liolaemus occipitalis presented a mean body temperature of 30.89 degrees C (SD = 4.43 degrees C; min = 16.4 degrees C; max = 40.2 degrees C; N = 270), that varied on a daily and seasonal basis according to microhabitat thermal alterations. The substrate temperature was the main heat source for thermoregulation of L. occipitalis as in all seasons of the year it was responsible for the animals' temperature variation (82% of the collected lizards in the spring; 60% in the summer; 84% in the fall and 68% in the winter). The results indicate that L. occipitalis is a saxicolous, thigmothermic and heliothermic species that regulates its body temperature through behavioral mechanisms. PMID- 17119844 TI - Reproduction of Chrotopterus auritus (Peters) in captivity (Chiroptera, Phyllostomidae). PMID- 17119845 TI - Life mapping: a 'Therapeutic Document' approach to needs assessment. AB - This study tested the effectiveness of The Life Map designed by the authors for holistic needs assessment for the promotion of mental health based on the World Health Organisation's six Quality of Life domains (1985). Tests were conducted of test-retest reliability and validity (criterion, discriminant, construct and concurrent) compared with an existing validated needs assessment, the Camberwell Assessment of Needs Inventory CAN-R. For main analyses, 43 mental health service users (SUs) were age-sex matched with 43 of non-service users (non-SUs). The Life Map significantly discriminated between these two study groups. Non-parametric correlations in each group showed significant relationship between a general quality of life question and the 'well-being' and the 'influence over' scores in each of the six domains. Consistent significant agreement was found in test retests conducted 2 weeks apart. Qualitative data in the Life Map was able to conceptualise the quantitative Life Map scores. This makes the Life Map a tool to improve communication between user and care-provider using a reliable felt and expressed needs assessment that empowers the service user with a 'voice' and empowers the care-provider with the capacity to compare aggregated needs between localities and for change over time. The limitations of this study, the Life Map's utility to facilitate meeting Standards 1 and 2 of the UK's National Service Framework for adult mental health and future directions are discussed. PMID- 17119846 TI - Determinants of health related quality of life and health state utility in patients with age related macular degeneration: the association of contrast sensitivity and visual acuity. AB - BACKGROUND: There has been increasing interest in the use of measures of health related quality of life (HRQoL) and health state utility values in Age Related Macular Degeneration (ARMD). Visual acuity has been found to be an important determinant of such measures in previous studies. More recently, another measure of visual impairment, contrast sensitivity has received considerable attention. We designed a study to examine whether the contribution of contrast sensitivity in explaining HRQoL and health utilities over and above that of visual acuity. METHODS: 209 patients with unilateral or bilateral ARMD were recruited into a cross-sectional study of patients from a large teaching hospital. Patients underwent visual tests (near and distant visual acuity, contrast sensitivity) and completed a vision function questionnaire, the VF-14, HUI3, and time trade-off. RESULTS: Using multivariate regression analysis, the study revealed that contrast sensitivity remained a statistically significant predictor of all outcome measures even when visual acuity was included. This result was supported by the correlation coefficients between measures. CONCLUSIONS: The measurement of contrast sensitivity appears to be better related to a person's HRQoL and health utility. Future studies should consider incorporating contrast sensitivity in addition to visual acuity. Studies, in particular economic evaluations, may underestimate the effect of treatment unless contrast sensitivity is considered. PMID- 17119847 TI - Health-related quality of life: Hepatocellular carcinoma, chronic liver disease, and the general population. AB - Health related quality of life (HRQL) has become an important endpoint in testing the efficacy of treatments for chronic liver disease (CLD) and the consequences of CLD which include hepatocellular carcinoma (HCC) and liver failure. However, a paucity of research on HRQL has been conducted with these patient populations. The aims of the present study were to compare persons diagnosed with HCC to persons diagnosed with CLD as well as with the general population (GP) on a disease-specific instrument measuring HRQL. If significant and clinically meaningful differences in HRQL exist, HRQL may be used as a corroborative indicator of disease progression in patients with CLD. Two hundred and seventy two people participated in the present study. Of these participants, 83 were diagnosed with HCC, 51 with CLD, and 138 were from the GP. None of the patients in the HCC or CLD samples were actively receiving chemotherapeutic treatments for the CLD or HCC. A sociodemographic questionnaire and the Functional Assessment of Cancer Therapy-Hepatobiliary (FACT-Hep) was administered to participants. The results of the study suggested that people diagnosed with HCC, prior to treatment, had a poorer overall HRQL when compared to those persons with CLD and the general population, as expected. The differences in HRQL were statistically significant as well as clinically meaningful. People diagnosed with CLD and HCC respectively, reported better social and family well-being than the general population. Furthermore, people with CLD reported equivalent emotional well-being as the general population sample. HRQL subscale scores, with the exception of social and family well-being, discriminated group membership. PMID- 17119848 TI - Enhanced NTPDase and 5'-nucleotidase activities in diabetes mellitus and iron overload model. AB - The activity of the enzymes NTPDase and 5'-nucleotidase was studied in both diabetes mellitus and an associated model of iron-overload. Rats were divided in five groups: citrate (CC), saline (S), diabetic (D), iron-overload (IO), and diabetic iron-overload (DIO). Diabetes was induced with alloxan (150 mg/kg), and iron-overload was induced with iron-dextran (10 intramuscular applications of +/ 80 mg/kg). The enzymatic activities were evaluated in the platelets. The results demonstrated an increase in the activity of NTPDase with substrates ATP and ADP (60% and 120%, respectively; P<0.001), and 5'-nucleotidase (60%, P<0.001). This increase was more intense in the IO and DIO groups. The results obtained in vitro showed an activation in ATP, ADP, and AMP hydrolysis between 1 microM and 1,000 microM ferric nitrate concentrations, being more pronounced at 100 microM and decreasing at 1,000 microM. We concluded that diabetes mellitus in association with iron-overload increased the hydrolysis of adenine nucleotides in platelets, contributing to the abnormalities found in these pathological conditions. PMID- 17119850 TI - Evaluation of SAT-1, SAT-2 and GalNAcT-1 mRNA in colon cancer by real-time PCR. AB - By qualitative and quantitative PCR, we evaluated the expression of three messengers coding for SAT-1, SAT-2 and GalNAcT-1 in human samples of intestinal cancer and some cell lines (breast cancer and melanomas). Qualitative PCR demonstrated, in human tissues but not in the cell lines examined, the presence of an mRNA that lacks hexon 3; experiments performed on transfected SKMEL-28 excluded a regulative role of this noncanonical mRNA. Data from real-time PCR, statistically analysed by ANOVA indicated that the mRNA expression of all the considered glycosyltransferases (SAT-1, SAT-2 and GalNAcT-1) was significantly different in tumours versus their own control. The ganglioside patterns in the examined samples did not correlate with mRNA expression; this finding demonstrates that ganglioside expression is the result of a very complex balance between anabolic and catabolic enzyme activities. Although this study is still preliminary, it opens a new possibility for neoplastic prognosis finding potential molecular markers among the mRNAs that codify for glycosyltransferases. PMID- 17119851 TI - Cloning and heterologous expression of bovine pyroglutamyl peptidase type-1 in Escherichia coli: purification, biochemical and kinetic characterisation. AB - We describe the cloning, expression and purification of the bovine XM866409 form of pyroglutamyl peptidase type-1 (PAP1). The cloned nucleotide sequence has an ORF coding for a primary sequence of 209 amino acid residues, which displays 98% identity with the human AJ278828 form of the enzyme. Three amino acid residues at positions 81, 205 and 208 were found to vary between the two sequences. The recombinant bovine PAP1 with a C-terminal His(6) tag (rBtaPAP1(6H)) was expressed in Escherichia coli XL10-Gold cells and purified by immobilised nickel ion affinity chromatography resulting in a yield of 2.6 mg of PAP1 per litre of culture. Purified rBtaPAP1(6H) had a specific activity of 3633 units mg(-1). SDS PAGE revealed a band for bovine PAP1 with a molecular weight of approximately 24 kDa, which is in good agreement with previously reported data on PAP1. The K (m) and k (cat) values obtained for rBtaPAP1(6H) were 59 muM and 3.5 s(-1), respectively. The optimum pH for activity was 9.0-9.5 and the optimum temperature was 37 degrees C. rBtaPAP1(6H) was found to have an absolute requirement for the thiol-reducing agent DTT, consistent with the expected property of a cysteine protease. Kinetic studies using the peptides pGlu-His-Pro-NH(2) (TRH), pGlu-Ala and pGlu-Val revealed K (i) values of 44.1, 141 and 652.17 microM, respectively. The lowest K (i), observed for Thyrotropin-releasing Hormone (TRH), indicates that rBtaPAP1(6H) has a higher affinity for tripeptides over dipeptides. PMID- 17119849 TI - Calcium signaling phenomena in heart diseases: a perspective. AB - Ca(2+) is a major intracellular messenger and nature has evolved multiple mechanisms to regulate free intracellular (Ca(2+))(i) level in situ. The Ca(2+) signal inducing contraction in cardiac muscle originates from two sources. Ca(2+) enters the cell through voltage dependent Ca(2+) channels. This Ca(2+) binds to and activates Ca(2+) release channels (ryanodine receptors) of the sarcoplasmic reticulum (SR) through a Ca(2+) induced Ca(2+) release (CICR) process. Entry of Ca(2+) with each contraction requires an equal amount of Ca(2+) extrusion within a single heartbeat to maintain Ca(2+) homeostasis and to ensure relaxation. Cardiac Ca(2+) extrusion mechanisms are mainly contributed by Na(+)/Ca(2+) exchanger and ATP dependent Ca(2+) pump (Ca(2+)-ATPase). These transport systems are important determinants of (Ca(2+))(i) level and cardiac contractility. Altered intracellular Ca(2+) handling importantly contributes to impaired contractility in heart failure. Chronic hyperactivity of the beta-adrenergic signaling pathway results in PKA-hyperphosphorylation of the cardiac RyR/intracellular Ca(2+) release channels. Numerous signaling molecules have been implicated in the development of hypertrophy and failure, including the beta adrenergic receptor, protein kinase C, Gq, and the down stream effectors such as mitogen activated protein kinases pathways, and the Ca(2+) regulated phosphatase calcineurin. A number of signaling pathways have now been identified that may be key regulators of changes in myocardial structure and function in response to mutations in structural components of the cardiomyocytes. Myocardial structure and signal transduction are now merging into a common field of research that will lead to a more complete understanding of the molecular mechanisms that underlie heart diseases. Recent progress in molecular cardiology makes it possible to envision a new therapeutic approach to heart failure (HF), targeting key molecules involved in intracellular Ca(2+) handling such as RyR, SERCA2a, and PLN. Controlling these molecular functions by different agents have been found to be beneficial in some experimental conditions. PMID- 17119853 TI - Use of porcine dermal collagen graft (Permacol) for hernia repair in contaminated fields. AB - BACKGROUND: Complicated hernias often involve contaminating surgical procedures in which the use of polypropylene meshes can be hazardous. Prostheses made of porcine dermal collagen (PDC) have recently been proposed as a means to offset the disadvantages of polypropylene meshes and have since been used in humans for hernia repairs. The aim of our study was to evaluate the safety and efficacy of incisional hernia repair using PDC as a mesh in complicated cases involving contamination. METHODS: A prospective study of hernia repair of complicated incisional hernias with contamination using PDC grafts was carried out at the Department of General, Emergency and Transplant Surgery of St Orsola-Malpighi University Hospital. RESULTS: From January 2004 up to the writing of this article, seven patients were treated for complicated incisional hernias with a PDC prosthesis. In six out of seven patients a bowel resection was carried out. There were not surgical complications. Morbidity was 14.2%. No recurrences and wound infections were observed. CONCLUSIONS: Incisional hernioplasty using PDC grafts is a potentially safe and efficient approach in complicated cases with contamination. PMID- 17119854 TI - "Suture hernia": identification of a new type of hernia presenting as a recurrence after laparoscopic ventral hernia repair. AB - After laparoscopic repair of ventral or incisional hernias, the recurrence rates reported are around 4%. Different mechanisms for the recurrences have been identified. We report two cases in which the patients were operated on laparoscopically for recurrence after laparoscopic ventral hernia repair. In both cases, the site of the recurrent hernia was situated at the transfascial fixation sutures. Patients were treated by laparoscopy with a larger intraperitoneal mesh covering the new hernia and the old mesh. PMID- 17119852 TI - A novel loss-of-function mutation in TP53 in an endometrial cancer cell line and uterine papillary serous carcinoma model. AB - The etiology of carcinoma of the uterine endometrium (ECa) is poorly understood. However, loss of apoptosis is one of the major factors that allow cancer cells to survive and progress. Hec50co, a poorly differentiated human ECa cell line, is widely used in the investigation of ECa. Previously, Hec50co xenograft tumor model in nude mice developed an advanced phenotype, similar to that of uterine papillary serous carcinoma (UPSC). Importantly, loss-of-function mutation in tumor suppressor TP53 was found in 20-30% of all ECa and >90% of UPSC. Thus, understanding the status of TP53 in Hec50co is essential for using Heco50co as a model for UPSC. To obtain an accurate genotype-phenotype status of TP53 in Hec50co, we performed mutation and functional analysis of TP53 gene of Hec50co by RT-PCR, genomic-PCR, and cloning and expression of mutant and wildtype TP53 alleles. We found a novel 42-bp deletion mutation in the exon6-intron6 splice junction of TP53 (TP53.del42bp) leading to a 113-bp exon6-deleted/skipped transcript was identified in Hec50co. In addition, the other TP53 allele in Hec50co is inactivated through a large deletion. Adenovirus (AD) harboring wildtype full-length TP53 cDNA induces caspase-dependent apoptosis; while the AD TP53.del42bp allele does not. In addition, messenger RNA of TP53.del42bp allele is stable whereas the protein product of TP53.del42bp allele is made but not stable. Taken together, we demonstrate that Hec50co is a TP53-null cell line possessing one TP53.del42bp allele and the other lost allele and therefore provides an excellent model to dissect the molecular and cellular bases of UPSC and other p53-null cancers. PMID- 17119855 TI - Laparoscopic retroperitoneal repair of a right-sided Bochdalek hernia. AB - Bochdalek hernias are rare congenital diaphragmatic defects. We report a case of a 50-year-old male with chronic shortness of breath who was diagnosed with a right-sided Bochdalek hernia. This hernia was repaired using a laparoscopic retroperitoneal approach. PMID- 17119856 TI - Endometriosis mimicking hernia recurrence. AB - Endometriosis is a common gynecologic condition and has been described in several locations, mostly in the pelvis. Extragenital endometriosis may appear as a painful nodule evoking an inguinal hernia. Scar endometriosis after inguinal hernia repair seems to be a rare occurrence. We report an unusual case of a 28 year-old woman who developed a scar endometriosis 2 years after an inguinal hernia repair. This case highlights that the presence of a painless inguinal mass similar to a recurrent hernia, with possible swelling related to the menstrual cycle, may evoke endometriosis, especially after a previous hernia repair and should lead to prompt diagnosis, wide excision, and gynecological advice. PMID- 17119857 TI - Could linear MRI measurements of hippocampus differentiate normal brain aging in elderly persons from Alzheimer disease? AB - Although mild progressive specific structural brain changes are commonly associated with normal human aging, it is unclear whether automatic or manual measurements of these structures can differentiate normal brain aging in elderly persons from patients suffering from cognitive impairment. The objective of this study was primarily to define, with a standard high resolution MRI, the range of normal linear age-specific values for the hippocampal formation (HF), and secondarily to differentiate hippocampal atrophy in normal aging from that occurring in Alzheimer disease (AD). Two MRI-based linear measurements of the hippocampal formation at the level of the head and of the tail, standardized by the cranial dimensions, were obtained from coronal and sagittal T1-weighted MR images in 25 normal elderly subjects, and 26 patients with AD. In this study, dimensions of the HF have been standardized and they revealed normal distributions for each side and each sex: the width of the hippocampal head at the level of the amygdala was 16.42 +/- 1.9 mm, and its height 7.93 +/- 1.4 mm; the width of the tail at the level of the cerebral aqueduct was 8.54 +/- 1.2 mm, and the height 5.74 +/- 0.4 mm. There were no significant differences in standardized dimensions of the HF between sides, sexes, or in comparison to head dimensions in the two groups. In addition, the median inter-observer agreement index was 93%. In contrast, the dimensions of the hippocampal formation decreased gradually with increasing age, owing to physiological atrophy, but this atrophy is more significant in the group of AD. PMID- 17119859 TI - The relationship between nail- and distal phalangeal bone involvement severity in patients with psoriasis. AB - We aimed to investigate the relationship between nail involvement and joint manifestations and whether there was a correlation between nail psoriasis severity and bone manifestations in psoriatic patients without symptomatic psoriatic arthritis in plaque type psoriasis. Thirty-one patients with nail involvement (16 men, 15 women, mean age 45.29+/-18.73) and 39 patients without nail involvement (16 men, 23 women, mean age 38.41+/-17.33) were enrolled in the study. X-ray of the hands and feet with magnification were performed. The distal interphalangeal (DIP) joint and bone (tuft of terminal phalanx) were evaluated. A scoring method was performed on the patients with nail involvement. There was no difference in DIP joint involvement in patients with or without finger- and toenail involvement (p=0.085 and p=0.062, respectively). However, the prevalence of bone involvement was higher in patients with finger- and toenail involvement than without finger- and toenail involvement (p=0.039 and p=0.021, respectively). A positive correlation was also determined between finger- and toenail psoriasis severity and bone involvement severity (r=0.379, p=0.001 and r=0.288, p=0.015). PMID- 17119858 TI - Kinking, coiling, and tortuosity of extracranial internal carotid artery: is it the effect of a metaplasia? AB - INTRODUCTION: Morphological anomalies of the extracranial internal carotid artery (ICA) cause symptomatic cerebrovascular insufficiency in 4-16% of the cases. The aim of the present study is to evaluate macroscopic and microscopic features of a group of extracranial ICA anomalies, specifically kinking, coiling, and tortuosity, eventually affecting the surgical approach. MATERIALS AND METHODS: From January 2003 to December 2005, 10 out of 169 (6%) revascularized patients (pts) were operated upon because of an ICA anomaly. They were all but two symptomatics. Seven pts were treated by ICA transection and end-to-side reimplantation of the ICA at the level of the carotid bulb; three pts were treated by ICA resection and end-to-end anastomosis. In all the cases a segment of ICA was resected; in three cases one more segment was also obtained from a common carotid artery (CCA) and these specimens were histologically examined. Patients were followed-up through a 3-year period. RESULTS: No pts died and none suffered of neurologic events. Duplex scan and arteriographic postoperative control showed the correct surgical reconstruction. Matching preoperative clinical findings with presence or absence of significant atherosclerotic stenotic lesion, we found out a positive cerebral CT in one pt (20%) in both groups; fluent neurological deficit was preeminent in pts with pure ICA anomalies (40% vs. 0%) (P = 0.2); pts with pure ICA anomalies were significantly younger than 65 years old (80% vs. 0%) (P = 0.03) and males were more involved by pure ICA anomalies (60% vs. 40%) (P = 0.1). The histological examination of ICA specimens showed a reduction of elastic fibers and muscular cells with a compensative increase of connective fibers. CONCLUSIONS: At our knowledge this is the first study focused on ICA anomalies like kinking, coiling, and tortuosity, comparing histologic features of CCA and ICA specimens coming from the same affected carotid axis. Our results, although preliminary, show elastic and muscular tissue substituted by loose connective tissue, configuring a metaplasia of tunica media limited to the ICA. Our hypothesis is that extracranial ICA, being a segment of transition between an elastic vessel (CCA) and a muscular vessel (intracranial ICA), is particularly subject to metaplastic transformation, analogously to other transition zones in human body. Our purpose is now to confirm by ultrastructural and molecular biology techniques, in a wider series, the presence of this metaplasia, since this could condition also the revascularization techniques. PMID- 17119860 TI - Harlequin ichthyosis and juvenile idiopathic arthritis: a rare combination. PMID- 17119861 TI - Anti-p53 antibodies in patients with dermatomyositis/polymyositis. AB - Dermatomyositis/polymyositis (DM/PM), which is often accompanied by various immunological abnormalities, was reported to be associated with an increased incidence of malignancies. In this study, we analyzed serum levels of anti-p53 antibody (anti-p53 Ab) in DM/PM patients and in normal controls. Serum levels of anti-p53 Abs were significantly higher in DM/PM patients than those in healthy controls. However, there was no significant difference between serum levels in patients with malignancies and those in patients without malignancies. Anti-p53 Abs were positive in 13% (4 out of 31) of the DM/PM patients. Of these four patients, only one had an internal malignancy. Immunoglobulin G levels were significantly higher in patients positive for anti-p53 Ab than those who were not. These results seemed to suggest that the presence of anti-p53 Abs in DM/PM patients is due to immunological abnormalities in this disease. PMID- 17119862 TI - 99mTechnetium pyrophosphate scintigraphy in the detection of skeletal muscle disease. AB - We aimed to assess the specificity and sensitivity of (99m)technetium pyrophosphate muscle scintigraphy in the diagnostic workup of patients with suspected myopathy. We reviewed the charts of 166 patients; 52% of the subjects had myalgias, 36% had muscle weakness, 45% had an elevated serum creatine kinase (CK), and 49% had an increased C reactive protein (CRP). Scintigraphy was positive in 34 patients (20%). The test was more sensitive in the presence of muscle weakness, elevated CK, or increased CRP. The presence of myalgias did not influence the odds. Sensitivity was 60% in patients with the final diagnosis of polymyositis, dermatomyositis, or inclusion body myositis, and 70% in noninflammatory myopathies. Eight percent had false positive scintigrams. In individuals with biopsy-proven myopathy (51 subjects), the diagnostic sensitivity was 43%, and its specificity was 60%. Low positive and high negative likelihood ratios (5.0 and 0.65, respectively) document an only limited diagnostic efficiency of (99m)Tc-PYP scintigraphy in the evaluation of inflammatory and noninflammatory myopathies and suggest that the test is not helpful in the routine diagnostic workup of muscle complaints, even after a priori selection of patients for CK plus CRP abnormalities. PMID- 17119863 TI - Quantitative ultrasound is better correlated with bone mineral density and biochemical bone markers in elderly women. AB - The association between quantitative ultrasound (QUS) and bone turnover in postmenopausal women of different ages is an area of continuous investigation. The aim of this study was to investigate the relationship of ultrasound parameters [broadband ultrasound attenuation (BUA) and speed of sound (SOS)] to bone mineral density (BMD) and biochemical markers of bone turnover in three age groups of postmenopausal women. One hundred and twenty-three postmenopausal Caucasian women were divided into three groups according to their age: group A, range 44-54 years, mean age (+/-SD) 48.3 +/- 2.3; group B, range 55-65 years, mean age 59.4 +/- 2.1; and group C, range 66-77 years, mean age 68.2 +/- 3.1. Ultrasound parameters were measured by the DTU-one imaging ultrasonometer in the calcaneus. BMD was assessed by dual-energy X-ray absorptiometry (DEXA) at the lumbar spine, femoral neck, and trochanter. Bone turnover was assessed by serum bone-specific alkaline phosphatase (BAP), urinary excretion of free deoxypyridinoline, N-telopeptides (NTX), and C-telopeptide breakdown products of type I collagen (CTX). QUS and BMD were significantly correlated in all sites, except hip BMD in group A. The most significant correlation was observed between BUA and femoral neck BMD in group C (r = 0.626, p < 0.01). BUA correlated significantly with BAP, NTX, and CTX (r = -0.434, -0.511, -0.478, respectively; p < 0.01), and SOS with BAP and NTX (r = -0.351 and -0.356, respectively; p < 0.05) only in group C. In groups A and B, ultrasound parameters did not correlate significantly to biochemical markers. Ultrasound parameters were better correlated to hip BMD and to biochemical markers of bone turnover in elderly postmenopausal women. These ultrasound measurements could be used as a screening test for bone status, either in nonambulatory third aged women or in those living in rural areas where attending medical centers with DEXA equipment and biochemical laboratories is difficult. PMID- 17119864 TI - Mixed connective tissue disease: a case with scleroderma renal crisis following abortion. AB - The term mixed connective tissue disease (MCTD) has been applied to a particular subset of patients with overlapping clinical features of systemic sclerosis, systemic lupus erythematosus, and polymyositis. Immune response to U1 ribonucleoprotein is the defining serological feature of MCTD. There are different organ and system involvements in MCTD including the heart, lung, kidney, muscle, joints, gastrointestinal, and hematologic involvements. Reports describing pregnancies in patients with MCTD are rare, and the results have been contradictory: a high risk of fetal loss and of disease exacerbation or no influence on fetus or mother. In MCTD, simultaneous pulmonary and renal involvement is very rare. In this paper, we report a case of MCTD with pulmonary involvement that developed scleroderma renal crisis after an abortion. PMID- 17119865 TI - Development of a microsatellite marker set applicable to genome-wide screening of cynomolgus monkeys (Macaca fascicularis). AB - To develop a microsatellite marker set applicable to genome-wide screening of cynomolgus monkeys (Macaca fascicularis), 148 microsatellite markers were selected from the human genome database. The polymorphisms and inheritance of PCR products were determined by screening twenty unrelated monkeys and by analysis of three families, respectively. As a result, 106 primers (72%) gave PCR products of the size expected for humans and rhesus monkeys. Among these products, polymorphism and single-gene inheritance in cynomolgus monkeys was observed for 66 markers (62%). The average number of alleles at the 66 polymorphic loci was 5.86 (range 2-10), and average heterozygosity was 0.63 (range 0.10-0.88). This is the first report of microsatellite markers for cynomolgus monkeys. Chromosomal mapping of these markers is now in progress. PMID- 17119866 TI - Do facial gestures, visibility or speed of movement influence gaze following responses in pigtail macaques? AB - This study investigated whether a human model's facial gestures, speed of head turn and visibility of face influenced gaze-following responses (GFR) in pigtail macaques. A human provided gaze cues by turning her head 90 degrees in one of four directions. Head turns were immediately followed by a facial movement (pucker, eyebrow raise, tongue protrusion, neutral), or were executed swiftly (<0.5 s), slowly (3 s) or whilst facing away from the monkeys. All monkeys reliably followed the gaze in all conditions with no differences between conditions. A greater frequency of GFR was found in females compared to males, and two hypotheses for this finding are discussed. PMID- 17119867 TI - Postglacial population expansion of Japanese macaques (Macaca fuscata) inferred from mitochondrial DNA phylogeography. AB - We investigated the diversity and phylogeography of mitochondrial DNA (mtDNA) in Japanese macaques (Macaca fuscata), an endemic species in Japan that has the northernmost distribution of any non-human primate species. DNA samples from 135 localities representing the entire range of this species were compared. A total of 53 unique haplotypes were observed for the 412-bp partial mtDNA control region sequence, with length variation distinguishing the two subspecies. Clustering analyses suggested two putative major haplogroups, of which one was geographically distributed in eastern Japan and the other in western Japan. The populations in the east showed lower mtDNA diversity than those in the west. Phylogeographical relationships of haplotypes depicted with minimum spanning network suggested differences in population structure. Population expansion was significant for the eastern but not the western population, suggesting establishment of the ancestral population was relatively long ago in the west and recent in the east. Based on fossil evidence and past climate and vegetation changes, we inferred that the postulated population expansion may have taken place after the last glacial period (after 15,000 years ago). Mitochondrial DNA showed contrasting results in both variability and phylogenetic status of local populations to those of previous studies using protein variations, particularly for populations in the periphery of the range, with special inference on habitat change during the glacial period in response to cold adaptation. PMID- 17119868 TI - Complications associated with neoadjuvant radiotherapy in the multidisciplinary treatment of retroperitoneal sarcomas. AB - INTRODUCTION: Retroperitoneal sarcomas (RPS) remain a therapeutic challenge due to high local recurrence rates. Preoperative RT offers theoretical advantages in the multidisciplinary care of RPS. The purpose of our study was to evaluate our experience using preoperative radiotherapy (PRT) in the treatment of RPS. METHODS: This is a single-institution review of patients with RPS treated with PRT from 1994 until 2004. Three radiation oncologists and four surgical oncologists were involved. Medical records, tumor registries, and death records were reviewed. RESULTS: Fourteen patients were included; nine were treated for primary presentation and five for recurrent disease. Histologic grade was grade I (n = 3), grade II (n = 3), and grade III (n = 8). Five patients received additional IORT. Radiotherapy complications were generally mild, including nausea (n = 3), diarrhea (n = 1), dehydration (n = 1), anemia (n = 1), and skin changes (n = 1); one required early cessation due to nausea. Thirteen patients had gross negative margins; while 7/13 had negative microscopic margins. Operative complications included anastomotic bleeding (n = 1), fluid collections (n = 2), ileus (n = 3), ascites (n = 2), temporary leg weakness (n = 1), and uncomplicated wound infections (n = 2). In patients with R0 or R1 resections, one and two year local control rates were 64 and 50%. Overall survival for all patients was 90% at 1 year and 74% at 2 years with median survival of 21 months. CONCLUSION: PRT and IORT can be administered effectively in carefully selected patients with resectable RPS. Larger multi-center studies are needed to delineate the role of PRT and IORT to improve local recurrence and survival rates in the treatment of RPS. PMID- 17119869 TI - Method of biopsy and incidence of positive margins in primary melanoma. AB - BACKGROUND: The staging of patients with primary melanoma is dependent on adequate sampling of the tumor thickness. Initial biopsies with a positive deep margin suggest inadequate sampling, potentially limiting accurate staging and affecting treatment decisions. METHODS: To determine the efficacy of shave biopsy to adequately sample the tumor, we retrospectively reviewed our pathology database for original pathology reports of primary melanomas accessioned between 01/01/04 and 6/30/05. The biopsies were evaluated by technique, the presence of tumor at the margins of the specimen, and specimen thickness. RESULTS: We identified 240 cases of primary melanoma; 223/240 were analyzable. The specimens were divided by biopsy technique (excisional, n = 51; punch, n = 44; and shave, n = 128). Shave and punch specimens had a significantly higher percentage of positive margins than excisional specimens (50, 68, and 16%, respectively; P < 0.0001). Shave specimens had a significantly higher percentage of positive deep margins than punch or excisional specimens (22, 7, and 2%, respectively; P = 0.0009). For melanomas or = 3.0, and 2.02 (1.31-3.11) for score > or = 3.5. In both instances the population fraction of the disease attributable to genetic predisposition exceeded 22%. Assuming a prior probability of at least 0.10 for the score to be predictive of coronary infarct risk, our findings are more likely than not to be truly positive. Our results, based on a simple score integrating the additive impact of 11 genetic polymorphisms, indicate that genetic predisposition accounts for a considerable fraction of the incidence of coronary infarct in the community. PMID- 17119880 TI - Investigating indicators and determinants of asthma in young adults. AB - BACKGROUND: In epidemiological studies on asthma determinants an extreme variability in results exists, probably due to different criteria utilised for defining of an asthma 'case' and for measuring determinants. We aimed to assess multiple indicators and multiple determinants of asthma in young adults by applying latent variable mixture models (LVMMs), a novel statistical modelling with hidden (or latent) variables. METHODS: We consider the pooled data of 1103 subjects (aged 20-44 years) from the three Italian centres of the European Community Respiratory Health Survey (ECRHS 1), a standardised database. Underlying multiple asthma indicators (clinicians' diagnosis, self-report symptoms, respiratory trials) both a latent two-class of asthma syndrome, and three continuous latent variables (severity of diagnosed asthma, severity of asthma symptoms, and severity of respiratory function) were investigated. RESULTS: Family history was the more relevant predictor of the two-class of asthma syndrome with a risk increase of about 60% per 1 relative with early life events (OR = 1.60, 95% CI: 1.30-1.97). Smoking, active and passive, are predictive for the indicators of severity of asthma symptoms. On average the risk increase of about 10% (OR = 1.10, 95%CI: 1.01-1.20) either per 1 source point of environmental tobacco smoke (ETS) or per 1 packet a day per 10 years. While, the risk of the indicators of both severity of asthma symptoms (OR = 1.59, 95%CI: 1.23-2.06) and severity of respiratory function (OR = 1.37, 95%CI: 1.03-1.82) increase in women compared to men, the risk of the indicators of severity of diagnosed asthma (OR = 0.57, 95%CI: 0.35-0.91) decreases. CONCLUSIONS: Considering latent modelling perspective for formulating plausible hypotheses in asthma research, this study highlighted that the host (genetic) component measured as number of relatives with life-events of asthma and/or allergies seems to be the primary determinants of overall observed asthma indicators summarised by hidden two-class of asthma syndrome. Furthermore, a secondary (or trigger) role of smoking on the continuous latent variable of severity of asthma symptoms, and a gender reversal effect were suggested. PMID- 17119881 TI - Adult height and lung function as markers of life course exposures: associations with risk factors and cause-specific mortality. AB - BACKGROUND AND OBJECTIVE: Pulmonary function and height may be regarded as adult indices of exposures accumulated across the entire life course and in early life, respectively. As such, we hypothesised that pulmonary function would be more strongly related to mortality than height. Studies of the association of height and lung function with mortality--which are currently modest in number--will clarify the relative utility of these risk indices and the mechanisms underlying observed patterns of disease risk. DESIGN, SETTING, AND PARTICIPANTS: Data were drawn from the Whitehall study, a prospective cohort study of 18,403 middle-aged non-industrial London-based male government employees conducted in the late 1960s. Data were collected on stature, spirometry measures (including forced expiratory volume in one second [FEV1]) and a range of covariates. These analyses are based on the 3083 non-smoking men with complete data. MAIN OUTCOME MEASURES: Mortality ascribed to all-causes, cardiovascular disease, respiratory disease and site-specific cancers. RESULTS: Both height and FEV1 were associated with a range of physiological, behavioural and socio-economic risk factors. Relations with these risk factors were seen more frequently for FEV1 and, where they occurred, were of somewhat higher magnitude. During a maximum of 35 years follow-up, half the non-smokers had died (n = 1545). FEV1 (HR(per one SD increase); 0.89; 0.84, 0.95) was somewhat more strongly related to total mortality than height (0.96; 0.91, 1.01) in a fully adjusted model, but this difference did not attain statistical significance at conventional levels (p-value for difference = 0.15). Of the eight independent disease-specific outcomes examined, the only convincing evidence of a differential effect was for deaths from respiratory causes which was unsurprisingly more strongly related to FEV1 than height (p-value for difference = 0.03). CONCLUSIONS: In the present study, height and FEV1 were essentially similarly related to both risk factors and mortality outcomes, thus not providing support for our hypothesis. Both factors would appear to have some utility as markers of early life exposures. PMID- 17119882 TI - [Liver resection in the state of Brandenburg]. PMID- 17119884 TI - [Response prediction--early response evaluation. Consequences for surgical oncology]. AB - Response to perioperative chemotherapy or chemoradiation can hardly be predicted on the basis of molecular marker analyses. In contrast, by means of metabolic and molecular imaging using positron emission tomography, response can be assessed as early as 14 days after the start chemotherapy for many tumors. There is no doubt that the prognosis of patients with surgically resected tumors is much better in the case of a response to chemotherapy or chemoradiation. Important consequences can be deduced from this regarding the indications for perioperative therapies, the radicality of surgery or the surgical indications per se. In the following, we delineate the current knowledge on response prediction and early response evaluation in selected tumor entities and draw possible conclusions for clinical practice and future clinical studies. PMID- 17119885 TI - [Advances in radio-oncology. From precision radiotherapy with photons to ion therapy with protons and carbon ions]. AB - Modern techniques in radiation oncology, such as fractionated stereotactic radiotherapy (FSRT), stereotactic radiosurgery (SRS) or intensity modulated radiotherapy (IMRT) allow the application of high local doses to defined treatment volumes, while normal structures in close vicinity can be spared; high local control rates can be achieved, while treatment-related toxicity can be minimized. Innovative Hi-Art tomotherapy systems offer an alternative, combining a 6 MV photon accelerator with a CT scanner. Ion beams, such as protons and carbon ions, have been shown to be beneficial for distinct tumor entities. Both offer a characteristic physical dose distribution with an inverse dose profile contributing to beneficial dose conformality. Carbon ions also offer the advantage of increased relative biological effectiveness. For certain tumor types, a significant increase in local control and survival rates could be obtained with carbon ions. PMID- 17119886 TI - [Minimal residual tumor in gastrointestinal carcinoma. Relevance to prognosis and oncologic surgical consequences]. AB - Isolated tumor cells as a consequence of minimal residual disease are often not detectable by routine diagnostic procedures. However, before or after surgery, isolated tumor cells in lymph nodes, the peritoneal cavity, blood, or bone marrow can frequently be identified by immunohistochemical or molecular methods. Failure to reveal the presence of such cells results in under-staging of tumor patients and may constitute the source of unexpected tumor recurrence after radical surgery. These facts emphasize the importance of isolated tumor cells at least as a surrogate marker. The frequency of appearance of isolated tumor cells in different organ systems also depends on the type of primary tumor. Developments in modern detection methods have led to increasing sensitivity but at the expense of specificity. Isolated tumor cells demonstrate remarkable heterogeneity with respect to proliferative potential and tumorigenicity. This characteristic is also reflected by a striking variability in the expression of various genes conditioning the aforementioned biological behavior. Unfortunately there is also remarkable heterogeneity in methods used for sampling and processing patient material as well as for the enrichment and detection of isolated tumor cells. Despite the ongoing controversies concerning detection methods and biological significance of isolated tumor cells, several clinical trials providing data supporting the prognostic relevance of minimal residual disease should also be considered for gastrointestinal carcinoma. In future this finding should be integrated in the planning of trials in surgical oncology, and "minimal residual disease" should receive stronger attention as a stratification criterion in such clinical studies. PMID- 17119887 TI - [Suicidology: suicide notes and their themes]. AB - BACKGROUND: The study of suicide notes has a long tradition. It is important to integrate this study into psychological autopsy and to make contact with the bereaved. For the first time, we present an Austrian-German study and its themes. MATERIAL AND METHODS: A consecutive series of suicides, investigated at the Department for Forensic Medicine in Vienna from Mai 2002 until April 2005, was examined for the presence of suicide notes. Suicide notes and demographic variables were integrated in an Access databank. RESULTS: A total of 29% of the those committing suicide left notes, however, the bereaved of only about a quarter allowed these to be studied. The major part was hand written, addressed to the general public or family, and shorter than 150 words. Frequent themes were specific instructions, emotions, themes regarding the environment such as the family, occupation and leisure time, and themes regarding stations of life and illness. PMID- 17119888 TI - [Cortical excitability in schizophrenia. Studies using transcranial magnetic stimulation]. AB - Abnormalities in brain plasticity, including abnormal information processing by disturbed cortical inhibition and facilitation in schizophrenia, have been described several times in the past. Transcranial magnetic stimulation (TMS) provides a neurophysiological technique for the measurement of cortical excitability, especially of the motoneural system. With TMS it is possible to explore important aspects of the pathophysiological mechanisms underlying schizophrenia using recently developed paradigms (paired pulse method, cortical silent period). This review summarizes the results of available diagnostic TMS studies in schizophrenia. Studies investigating the efficacy of therapeutic repetitive magnetic stimulation in patients with treatment resistant hallucinations or predominantly negative symptoms were not considered. The reviewed studies support the assumption of reduced cortical inhibition, probably caused by GABAergic deficit, in schizophrenia. Factors influencing the study results, the limitations of this promising technique in schizophrenia, and further research options are discussed. PMID- 17119889 TI - [Mixed affective disorders]. AB - The Kraepelin concept of "mixed states" has experienced a renaissance over the last two decades. This has been caused by clinical as well as theoretically relevant factors. Of particular clinical relevance is the fact that more than 40% of all patients with bipolar disorders show at least one mixed affective episode during the course of their illness. However, the correct assessment and the precise classification of the symptoms are very important. A diagnosis according to the tight criteria of ICD-10 or DSM-IV, or perhaps based on the moderate Pisa or Cincinnati criteria, is recommended, whereas the use of broader definitions cannot be advised. Mixed schizoaffective episodes are under-diagnosed in comparison to pure affective mixed episodes, although both are defined by ICD-10 as well as by DSM-IV, and the frequency of occurrence is almost the same. Mixed schizoaffective episodes appear to be one of the most severe forms of bipolar disorders. Their clinical relevance is mainly caused by their unfavourable prognosis and difficulty in treatment. Atypical neuroleptics combined with anticonvulsives have proved to be more effective than any other psychopharmacological substances. The occurrence of mixed depressive and manic symptoms during one and the same episode is theoretically important, especially in the sense of their nosological and etiological differentiation. PMID- 17119890 TI - [Multiaxial classification of stalking. Guidelines for the assessment of criminal liability and prognosis]. AB - Stalking is a widespread phenomenon describing a pattern of intrusive and threatening behaviour that leads to the victim's perception of being harassed and of him or her being rendered fearful. Physical assault and even homicide may occur in the context of stalking. Anglo-Saxon studies have revealed a lifetime prevalence of being a victim of stalking ranging from 4-7% in men and 12-17% in women. Recently, these rates have been confirmed by the first community based study carried out in Germany. As a stalker can have a number of victims during his or her lifetime, the prevalence of stalkers may be less than this, although at present data for this are lacking. Although the phenomenology of stalking appears to be rather homogenous, fairly distinct stalker typologies and perpetrator-victim relationships have to be considered. Requests for psychiatric and forensic assessment of stalkers are increasing. According to the German penal code, psychiatrists must provide expert opinion on criminal responsibility and the placement of stalkers. So far, all typologies of stalkers refer to the Anglo Saxon cultural background and do not consider the special needs of German forensic psychiatry. In particular, the psychopathological dimension is widely neglected in common typologies. The present paper proposes a multiaxial typology of stalking that considers the psychopathological dimension, the relationship between stalker and victim and motivational aspects. Consequences for the forensic psychiatric assessment according to section 20, 21 StGB are outlined. It should be pointed out that stalking is not a new diagnostic category, but only involves, at a descriptive level, deviation from a normal behavioural pattern. The central components of the forensic psychiatric assessment remain the known diagnostic categories, the effects of which on behaviour can be analysed. PMID- 17119891 TI - [Endovascular treatment for intracranial stenoses. A common statement by neurologists and neuroradiologists]. AB - Intracranial stenoses cause 5-10% of all strokes and are increasingly detected by means of modern imaging methods. The stroke danger of high-grade symptomatic stenoses is relatively high, with an annual risk of approximately 10% under medical treatment. Coumadin increases the risk of hemorrhage, and after risk/benefit considerations, antiplatelets should be preferred for antithrombotic therapy. Despite optimized medical treatment, a small group of patients with recurrent symptoms or symptomatic stenoses without adequate collateral supply probably carry higher spontaneous stroke risk and may be considered for intracranial stenting, which itself is associated with procedural risks of up to 10%. Currently published case series show relatively high complication rates as a major drawback of endovascular treatment, mainly strokes after occlusion of perforating branches extending from the stenotic vessel segment or hemorrhagic complications. According to data from smaller feasibility studies, stroke rates in follow-up after successful stenting seem to be low. The average rate of high grade restenosis with possible indication for reintervention is 10%. Improvements in endovascular treatment aim at reducing vessel wall trauma during balloon angioplasty by underdilatation or the use of self-expanding stents. Until complication rates are dependably reduced to values of 5-6%, indication for endovascular treatment should be restricted to patients without therapeutic alternatives. According to limited data with large variation between different studies, a prospective multicentric registry is proposed for systematic evaluation and further development of the method. PMID- 17119892 TI - ["Hypermetamorphosis". Heinrich Neumann's (1814-1884) legacy]. AB - The term "hypermetamorphosis" was originally coined in 1859 by an almost forgotten psychiatrist from Breslau, now Wroclaw in Poland, Heinrich Wilhelm Neumann. The 1906 textbook of his assistant Wernicke transmitted the concept to Kluver and Bucy, who understood it as the "excessive tendency to take notice of and to attend and react to every visual stimulus" in their syndrome description of bitemporal lobectomy in the monkey (1937-1939). Hypermetamorphosis so far has not been properly operationalized, and the concept appears outdated. Components such as "compulsive manipulation", "magnet reaction/groping", "compulsive grasping/grasp reflex", "utilization behavior", and "environmental dependency syndrome" can be better delineated and are commonly seen in frontal rather than temporal lesions. They may occur with frontal contusion, anterior cerebral artery infarction, in diseases of the Pick complex, and basal ganglia neurodegeneration. PMID- 17119893 TI - [Radiological imaging of the upper gastrointestinal tract. Part 1. The esophagus]. AB - In the diagnosis of diseases of the esophagus, conventional x-ray evaluation still plays a more important role than endoscopy in the visualization of stenoses. CT plays a major role in the staging of malignancies of the esophagus, while MRI plays does not play a major part in the diagnostic evaluation of the upper GI-tract but is equal to CT for the staging and evaluation of the extent of local infiltration. The main indication for the radiological examination of the esophagus by barium studies is dysphagia. The use of barium allows a functional examination of esophageal motility. Swallow motility disorders can be diagnosed by videofluorography using high frame rate imaging. Zenker's diverticulum and other pulsion diverticula should also be investigated by functional esophageal imaging. Candida esophagitis can be identified by its characteristic ulcerations using barium swallow. The extension of gastroesophageal hernias are more accurately evaluated with barium studies than with endoscopy. The diagnosis of gastroesophageal reflux disease should be made by barium studies, but discrete inflammation as well as epithelial dysplasia are best investigated by classic endoscopy and modern endoscopic techniques. In cases of esophageal carcinoma, radiology adds to the findings of endoscopy and endosonography. PMID- 17119894 TI - [Imaging of spinal tumors]. AB - Spinal tumors are often categorized into extradural, intradural extramedullary, or intramedullary. Although this classification represents somewhat of an overgeneralization as a lesion may reside in two compartments, it still helps to characterize spinal tumors. In the intradural, extramedullary space, primary tumors, such as neurofibroma and meningioma, are relatively common. Secondary tumors or leptomeningeal enhancement also occur. In the intramedullary space, primary tumors are far more common than secondary tumors or metastases. PMID- 17119896 TI - [Evidence-based pharmacological metaphylaxis of stone disease]. AB - Approximately one-fourth of urolithiasis patients are at high risk for recurrent stone formation or severe metabolic disturbances. These patients need specific metaphylaxis for effective stone prevention, adjusted to their individual metabolic risk. Recent recommendations for the pharmacological treatment of stone diseases are summarized in this article. For the different treatment options, evidence from the literature was assessed. In addition, a follow-up concept for pharmacologically treated high-risk stone formers is discussed. PMID- 17119897 TI - [HIV-related symptoms]. AB - The course of HIV-infection has changed dramatically since the beginning of the epidemic. Opportunistic infections and AIDS-defining tumors are diagnosed less frequently since the introduction of antiretroviral therapy and the time of survival has increased. According to German-Austrian therapy guidelines highly active antiretroviral therapy (HAART) should be started on the onset of HIV related symptoms and/or when the CD4 cell count is lower than 350/microl. Patients should be treated in specialized centres because of the complexity of HIV-infection and its therapy. For monitoring CD4 cell counts and viral load are determined. Reasons for therapeutic failure can be drug interactions, resistance or compliance problems. Although HIV-infection is often compared to insulin dependent diabetes mellitus psychological and social impact on HIV patients is still high. Increasing viral multi-drug resistance, long-term toxicity like lipodystrophy, osteoporosis and cardiovascular disease are only some problems HIV infected patients are facing in the next years. In Germany 600 to 700 patients still die because of AIDS every year. PMID- 17119898 TI - [Laboratory diagnostics of systemic autoimmune diseases. Part 1. Collagenoses]. AB - This is the first part of a series of articles on the laboratory diagnostics of rheumatic diseases and will consider the systemic autoimmune diseases lupus erythematosus, Sjogren's syndrome, systemic sclerosis, dermato/polymyositis and mixed connective tissue disease (MCTD, SHARP syndrome). The basis for diagnostics is the presence of antinuclear antibodies (ANA). Initially, these antibodies are detected using a screening test. This must be followed by the identification of the patient's individual autoantibody specificities, which then yields important diagnostic clues. Disease activity may be monitored serologically by following the titers of selected autoantibodies and, in certain patients, by examining complement consumption. PMID- 17119899 TI - [Ankylosing spondylitis--current state of imaging including scoring methods]. AB - Conventional radiography and magnetic resonance imaging (MRI) are currently the most widely used imaging methods for the initial diagnostic evaluation and follow up of patients with ankylosing spondylitis (AS). Scintigraphy, computed tomography (CT), and positron emission tomography (PET) only play minor roles, although some are being further developed. AS is characterized by inflammatory changes to the sacroiliac joints (SIJs) and spine, as well as asymmetrical arthritis of the peripheral joints and joints near the trunk. The diagnosis of AS is based on clinical parameters and the presence of chronic inflammatory changes to the SIJs on conventional radiographs. Typical radiographic changes also involve the spine. MRI depicts not only chronic changes, but also active inflammatory lesions, which are important for the diagnosis of early disease and precursors of AS. The scoring system of choice for quantifying spinal changes depicted by conventional radiography is the modified Stoke Ankylosing Spondylitis Spine Score (mSASSS). MRI allows the quantitative evaluation of changes involving the SIJs and the spine. Various MRI scoring systems have been proposed to quantify these changes, but they require further validation. This review article presents the imaging modalities used in AS patients, typical findings, and relevant methods of analysis. The most recent developments are discussed. PMID- 17119901 TI - Glioblastoma--the consequences of advanced patient age on treatment and survival. AB - Glioblastoma is the most common primary brain tumor. Recent evidence suggests that aggressive treatment is also effective in elderly patients. However, large patient series are missing. The aim of this retrospective study was to determine prognostic factors in a large series (n=345) of elderly patients surgically treated for newly diagnosed glioblastoma (WHO grade IV) at a single institution between 1991 and 2002. U-tests (Mann Whitney), chi-square tests, log-rank tests/Kaplan-Meier plots and Cox regression models were used for statistical analysis. Based on the maximum difference in median survival, a threshold of 60 years was used to separate younger from older patients. In total, 185 patients (53.6%) were over 60 years old. In these individuals, total tumor resection, radiotherapy and reoperation for tumor recurrence were identified as independent prognostic factors. When total surgical resection was combined with radiotherapy and reoperation, Kaplan-Meier analysis revealed a median survival of up to 64 weeks in elderly patients. Our data indicate that total tumor resection, radiotherapy and reoperation should also be considered in selected elderly patients. Age alone should not generally exclude elderly individuals from aggressive treatment. PMID- 17119900 TI - [Recommendations for the management of ankylosing spodylitis after ASAS/EULAR. Evaluation in the German language area]. AB - AIM: Our aim was to adapt and implement the evidence based recommendations for the management of ankylosing spodylitis (AS) of the "Assessments in AS" (ASAS) International Working Group together with the European League Against Rheumatism (EULAR) within the framework of a competence network (CN) in rheumatology in the German language area. METHODS: The ASAS/EULAR project calculated the effective size (ES), rate ratio, number of patients requiring treatment (number needed to treat, NNT) and the incremental cost-effectiveness ratio (ICER). The strength of the recommendations was determined through the evidence level found in the literature, the risk-benefit trade-off and the clinical experience of the experts. The recommendations were recently published in English. All of the centers taking part in the study area Spondyloarthritis (SpA) CN, as well as an additional 35 experts, were sent the English manuscript. All 35 participants were asked to evaluate the ten main management recommendations on a scale from 0 to 10. RESULTS: The recommendations encompass the use of drugs such as non-steroid anti-inflammatories (NSAR), which, along with conventional NSAR include coxibs and the parallel application of gastroprotectives, so called disease-modifying anti-rheumatic drugs, biologicals, simple analgesics, local and systematic glucocorticoids, non-drug therapies (such as patient training, medical training therapy and physiotherapy), in addition to surgical treatment methods. Moreover, three general recommendations were formulated and a therapy scheme created, taking into consideration the various clinical manifestations. The strength of the ASAS/EULAR recommendations was generally high. There was a marked consensus between the German speaking experts and the international proposal: a mean of 9.13 with relatively low variation between the recommendations. SUMMARY: Ten key recommendations for the treatment of AS were developed. These were strengthened by a systematic search of the literature and by expert consensus. The large group of German speaking experts were largely in agreement with the proposal. This can be seen as a starting point for the dissemination and implementation of the recommendations. PMID- 17119902 TI - Computational modeling of arterial wall growth. Attempts towards patient-specific simulations based on computer tomography. AB - The present manuscript documents our first experiences with a computational model for stress-induced arterial wall growth and in-stent restenosis related to atherosclerosis. The underlying theoretical framework is provided by the kinematics of finite growth combined with open system thermodynamics. The computational simulation is embedded in a finite element approach in which growth is essentially captured by a single scalar-valued growth factor introduced as internal variable on the integration point level. The conceptual simplicity of the model enables its straightforward implementation into standard commercial finite element codes. Qualitative studies of stress-induced changes of the arterial wall thickness in response to balloon angioplasty or stenting are presented to illustrate the features of the suggested growth model. First attempts towards a patient-specific simulation based on realistic artery morphologies generated from computer tomography data are discussed. PMID- 17119903 TI - Efficient solubilization, purification of recombinant extracellular alpha-amylase from pyrococcus furiosus expressed as inclusion bodies in Escherichia coli. AB - The gene encoding the Pyrococcus furiosus extracellular alpha-amylase (PFA) was amplified by PCR from P. furiosus genomic DNA and was highly expressed in Escherichia coli BL21-Codon Plus (DE3)-RIL. The recombinant alpha-amylase was mainly expressed in the form of insoluble inclusion bodies. An improved purification method was established in this paper. The solubilization of the inclusion bodies was achieved by 90 degrees C treatment for 3 min in Britton Robinson buffer at pH 10.5. The solubilized PFA was then diluted and subsequently purified by Phenyl Sepharose chromatography. The overall yield of the new purification method was about 58,000 U/g wet cells, which is higher than previously reported. PMID- 17119904 TI - Fast chemical shift mapping with multiecho balanced SSFP. AB - OBJECT: A method is proposed that provides spectroscopic images with high spatial resolution and moderate spectral resolution at very short total data acquisition times. MATERIALS AND METHODS: Balanced steady-state free precession (bSSFP, TrueFISP, FIESTA, b-FFE) is combined with a multiecho readout gradient and frequency-sensitive reconstruction such as Fourier reconstruction known from echo planar spectroscopic imaging (EPSI) or matrix inversion. Balanced SSFP imaging requires short repetition times to minimize banding artefacts, thereby restricting the achievable frequency resolution. RESULTS: Two-dimensional (2D) high-resolution spectroscopic images were produced of three 1H resonances (water, acetone and fat) on phantoms and water/fat separation in vivo within 1-2 s. Additionally, fast 31P spectroscopic images were acquired from a phantom consisting of two resonances within 195 ms. CONCLUSION: Frequency-sensitive reconstruction of multiecho bSSFP data can provide spectroscopic images with high spatial and temporal resolution while the frequency resolution is moderate at around 100 Hz. The method can also separate more than three resonances, allowing for hetero-nuclei metabolite mapping, for example 13C and 31P. PMID- 17119905 TI - CaMKII, an emerging molecular driver for calcium homeostasis, arrhythmias, and cardiac dysfunction. AB - Maintenance of cytoplasmic calcium homeostasis is critical for all cells. An exciting field has emerged in elucidating the multiple roles that Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) plays in regulating Ca(2+) cycling in normal cardiac myocytes and in pathophysiological states. Moreover, CaMKII was recently identified as a potential drug target in cardiac disease. This work has given us a closer view of the complexity and therapeutic possibilities of CaMKII regulation of Ca(2+) signaling in cardiac myocytes. PMID- 17119906 TI - Characterization of novel missense mutations in CYP21 causing congenital adrenal hyperplasia. AB - Congenital adrenal hyperplasia due to 21-hydroxylase deficiency is the most common inherited disorder of steroid metabolism, with an incidence of 1/10,000 in the general Caucasian population. Although most patients carry a deletion of the CYP21 gene or any of nine pseudogene-derived point mutations, the number of reported rare mutations continues to increase, and consist today of more than 80 different point mutations. In this study, we report the characterization of four additional missense mutations in CYP21. Two of these, L166P and A391T, are novel missense mutations, whereas the R479L and R483Q mutations have been detected previously. Functional assays of mutagenized CYP21 were performed in transiently transfected mammalian cells in vitro, and enzymatic ability of substrate conversion of the two natural substrates of CYP21-17-hydroxyprogesterone and progesterone-was determined. All mutants displayed reduced in vitro enzyme activities compared with wild type, but to different extents, corresponding to clinical phenotypes that span the whole spectrum of disease severity. Functional studies are important to further establish the relationships between genotype and clinical phenotype as well as in vitro CYP21 activity in congenital adrenal hyperplasia due to 21-hydroxylase deficiency. This has relevance for diagnosis, prognosis, and genetic counseling for affected families. PMID- 17119907 TI - Geographic variations in seed dispersal by ants: are plant and seed traits decisive? AB - The effect of local ant species on the dispersal success of a myrmecochorous plant, Helleborus foetidus, was analyzed in two populations of the Iberian Peninsula (Caurel and Cazorla, respectively). The contribution of the various local ant species to dispersal was very unequal. While 5 and 19 ant taxa visited the plants of Caurel and Cazorla, respectively, most removal activity (67 and 80%) was performed by two species only (Formica lugubris and Camponotus cruentatus, respectively). Visits by dispersers were also unequally distributed between neighboring plants. While some plants were always visited during the period of seed release, others were never visited. A regression model indicated that this pattern might be explained by two plant traits: ants preferred to visit plants that released more seeds and whose elaiosomes were richer in oleic acid. Although it has long been known that this compound triggers removal by ants, it is the first demonstration that quantitative variations in elaiosome traits contribute to variation in dispersal success. Finally, other variables being equal, morphological traits (seed size, elaiosome size, and elaiosome/seed size ratio) did not affect ant behavior. Although myrmecochory has long been considered a diffuse interaction, our results support the idea that, at local scale, a limited number of ant species may be decisive to its evolution. PMID- 17119908 TI - Parasitoid-induced mortality of Araneus omnicolor (Araneae, Araneidae) by Hymenoepimecis sp. (Hymenoptera, Ichneumonidae) in southeastern Brazil. AB - All species included in the Polysphincta genus-group develop as ectophagous parasitoids of active spiders, killing their hosts prior to pupation. However, little information regarding natural history and ovipositing behavior of most species are available. In this study we inspected 85 webs of Araneus omnicolor to evaluate the frequency of parasitism and host size preferences of the wasp Hymenoepimecis sp. We also described the web characteristics of normal and parasitized spiders and the wasp ovipositing behavior. About 41% of the adult females of A. omnicolor inspected were parasitized. The highest incidence of parasitism was observed among relatively small females while no egg or larva was found in large individuals. Araneus omnicolor builds a strong web composed of an orb and barrier threads, where the spider rests within a curled leaf. The parasitoid larva builds its cocoon within this refuge, and modified cocoon webs were not observed. The ovipositing behavior of Hymenoepimecis sp. was very similar to that of Hymenoepimecis argyraphaga parasitizing Leucauge argyra, including the position of the sting, the killing of a previously attached larva, and the expelling of the egg from the base of the ovipositor. PMID- 17119909 TI - Floral odor learning within the hive affects honeybees' foraging decisions. AB - Honeybees learn odor cues quickly and efficiently when visiting rewarding flowers. Memorization of these cues facilitates the localization and recognition of food sources during foraging flights. Bees can also use information gained inside the hive during social interactions with successful foragers. An important information cue that can be learned during these interactions is food odor. However, little is known about how floral odors learned in the hive affect later decisions of foragers in the field. We studied the effect of food scent on foraging preferences when this learning is acquired directly inside the hive. By using in-hive feeders that were removed 24 h before the test, we showed that foragers use the odor information acquired during a 3-day stimulation period with a scented solution during a food-choice situation outside the nest. This bias in food preference is maintained even 24 h after the replacement of all the hive combs. Thus, without being previously collected outside by foragers, food odors learned within the hive can be used during short-range foraging flights. Moreover, correct landings at a dual-choice device after replacing the storing combs suggests that long-term memories formed within the colony can be retrieved while bees search for food in the field. PMID- 17119910 TI - Living in a "stethoscope": burrow-acoustics promote auditory specializations in subterranean rodents. AB - Subterranean mammals rely to a great extent on audition for communication and to be alerted to danger. The only hitherto published report on burrow acoustics revealed that in tunnels of blind mole-rats (Spalax ehrenbergi), airborne sounds of 440 Hz propagated best whereas lower and higher frequencies were effectively attenuated. Morpho-functional analyses classify the ear of subterranean mammals as a low-sensitivity and low-frequency device. Concordantly, hearing is characterized by low sensitivity and a restricted frequency range tuned to low frequencies (0.5-4 kHz). Some authors considered the restricted hearing in subterranean mammals vestigial and degenerate due to under-stimulation. In contrast to this view stand a rich (mostly low-frequency) vocal repertoire and progressive structural specializations of the middle and inner ear. Thus, other authors considered these hearing characteristics adaptive. To test the hypothesis that acoustical environment in burrows of different species of subterranean mammals is similar, we measured sound attenuation in burrows of Fukomys mole-rats (formerly known as Cryptomys, cf. Kock et al. 2006) of two differently sized species at different locations in Zambia. We show that in these burrows, low frequency sounds (200-800 Hz) are not only least attenuated but also their amplitude may be amplified like in a stethoscope (up to two times over 1 m). We suggest that hearing sensitivity has decreased during evolution of subterranean mammals to avoid over-stimulation of the ear in their natural environment. PMID- 17119911 TI - Extensive macrosynteny between Medicago truncatula and Lens culinaris ssp. culinaris. AB - The first predominantly gene-based genetic linkage map of lentil (Lens culinaris ssp. culinaris) was constructed using an F5 population developed from a cross between the cultivars Digger (ILL5722) and Northfield (ILL5588) using 79 intron targeted amplified polymorphic (ITAP) and 18 genomic simple sequence repeat (SSR) markers. Linkage analysis revealed seven linkage groups (LGs) comprised of 5-25 markers that varied in length from 80.2 to 274.6 cM. The genome map spanned a total length of 928.4 cM. Clear evidence of a simple and direct macrosyntenic relationship between lentil and Medicago truncatula was observed. Sixty-six out of the 71 gene-based markers, which were previously assigned to M. truncatula genetic and physical maps, were found in regions syntenic between the Lens c. ssp. culinaris and M. truncatula genomes. However, there was evidence of moderate chromosomal rearrangements which may account for the difference in chromosome numbers between these two legume species. Eighteen common SSR markers were used to connect the current map with the most comprehensive and recent map that exists for lentil, providing the syntenic context of four important domestication traits. The composite map presented, anchored with orthologous markers mapped in M. truncatula, provides a strong foundation for the future use of genomic and genetic information in lentil genetic analysis and breeding. PMID- 17119912 TI - QTL associated with horizontal resistance to soybean cyst nematode in Glycine soja PI464925B. AB - Soybean cyst nematode (Heterodera glycines Ichinohe; SCN) is the primary disease responsible for yield loss of soybean [Glycine max (L.) Merr.]. Resistant cultivars are an effective management tool; however, the sources currently available have common resistant genes. Glycine soja Sieb. and Zucc., the wild ancestor of domesticated soybean, represents a diverse germplasm pool with known SCN resistance. The objectives of this research were to: (1) determine the genetic variation and inheritance of SCN resistance in a G. max ('S08-80') x G. soja (PI464925B) F (4:5) recombinant inbred line (RIL) population; and (2) identify and evaluate quantitative trait loci (QTL) associated with SCN resistance. Transgressive segregation for resistance was observed, although neither parent was resistant to the Chatham and Ruthven SCN isolates. Broad sense heritability was 0.81 for the Ruthven and 0.91 for the Chatham isolate. Root dry weight was a significant covariate that influenced cyst counts. One RIL [female index (FI) = 5.2 +/- 1.11] was identified as resistant to the Chatham isolate (FI < 10). Seventeen and three RILs infected with Chatham and Ruthven isolates, respectively, had mean adjusted cyst counts of zero. Unique and novel QTL, which derived resistance from G. soja, were identified on linkage groups I, K, and O, and individually explained 8, 7 and 5% (LOD = 2.1-2.7) of the total phenotypic variation, respectively. Significant epistatic interactions were found between pairs of SSR markers that individually may or may not have been associated with SCN resistance, which explained between 10 and 15% of the total phenotypic variation. Best-fit regression models explained 21 and 31% of the total phenotypic variation in the RIL population to the Chatham and Ruthven isolates, respectively. The results of this study help to improve the understanding of the genetic control of SCN resistance in soybean caused by minor genes resulting in horizontal resistance. The incorporation of the novel resistance QTL from G. soja could increase the durability of SCN-resistance in soybean cultivars, especially if major gene resistance breaks down. PMID- 17119913 TI - Modeling segregation distortion for viability selection. I. Reconstruction of linkage maps with distorted markers. AB - Molecular markers have been widely used to map quantitative trait loci (QTL). The QTL mapping partly relies on accurate linkage maps. The non-Mendelian segregation of markers, which affects not only the estimation of genetic distance between two markers but also the order of markers on a same linkage group, is usually observed in QTL analysis. However, these distorted markers are often ignored in the real data analysis of QTL mapping so that some important information may be lost. In this paper, we developed a multipoint approach via Hidden Markov chain model to reconstruct the linkage maps given a specified gene order while simultaneously making use of distorted, dominant and missing markers in an F(2) population. The new method was compared with the methods in the MapManager and Mapmaker programs, respectively, and verified by a series of Monte Carlo simulation experiments along with a working example. Results showed that the adjusted linkage maps can be used for further QTL or segregation distortion locus (SDL) analysis unless there are strong evidences to prove that all markers show normal Mendelian segregation. PMID- 17119914 TI - Impact of the population at risk of diabetes on projections of diabetes burden in the United States: an epidemic on the way. AB - AIMS/HYPOTHESIS: The aim of this study was to make projections of the future diabetes burden for the adult US population based in part on the prevalence of individuals at high risk of developing diabetes. MATERIALS AND METHODS: Models were created from data in the nationally representative National Health and Nutrition Examination Survey (NHANES) II mortality survey (1976-1992), the NHANES III (1988-1994) and the NHANES 1999-2002. Population models for adults (>20 years of age) from NHANES III data were fitted to known diabetes prevalence in the NHANES 1999-2002 before making future projections. We used a multivariable diabetes risk score to estimate the likelihood of diabetes incidence in 10 years. Estimates of future diabetes (diagnosed and undiagnosed) prevalence in 2011, 2021, and 2031 were made under several assumptions. RESULTS: Based on the multivariable diabetes risk score, the number of adults at high risk of diabetes was 38.4 million in 1991 and 49.9 million in 2001. The total diabetes burden is anticipated to be 11.5% (25.4 million) in 2011, 13.5% (32.6 million) in 2021, and 14.5% (37.7 million) in 2031. Among individuals aged 30 to 39 years old who are not currently targeted for screening according to age, the prevalence of diabetes is expected to rise from 3.7% in 2001 to 5.2% in 2031. By 2031, 20.2% of adult Hispanic individuals are expected to have diabetes. CONCLUSIONS/INTERPRETATION: The prevalence of diabetes is projected to rise to substantially greater levels than previously estimated. Diabetes prevalence within the Hispanic community is projected to be potentially overwhelming. PMID- 17119915 TI - Comment on: Brugman S et al. (2006) Antibiotic treatment partially protects against type 1 diabetes in the Bio-Breeding diabetes-prone rat. Is the gut flora involved in the development of type 1 diabetes? Diabetologia 49:2105-2108. PMID- 17119916 TI - Exercise under hyperinsulinaemic conditions increases whole-body glucose disposal without affecting muscle glycogen utilisation in type 1 diabetes. AB - AIMS/HYPOTHESIS: We examined whole-body and muscle metabolism in patients with type 1 diabetes during moderate exercise at differing circulating insulin concentrations. METHODS: Eight men (mean +/- SEM age 36.4 +/- 1.5 years; diabetes duration 11.3 +/- 1.4 years; BMI 24.6 +/- 0.7 kg/m(2); HbA(1c) 7.9 +/- 0.2% and VO(2) peak 44.5 +/- 1.2 ml kg(-1) min(-1)) with type 1 diabetes were studied on two occasions at rest (2 h) and during 45 min of cycling at 60% maximum VO(2) with insulin infused at the rate of either 15 (LO study) or 50 (HI) mU m(-2) min( 1) and blood glucose clamped at 8 mmol/l. Indirect calorimetry, insulin-glucose clamps and thigh muscle biopsies were employed to measure whole-body energy and muscle metabolism. RESULTS: Fat oxidation contributed 15 and 23% to total energy expenditure during exercise in the HI and LO studies, respectively. The respective carbohydrate (CHO) oxidation rates were 31.7 +/- 2.7 and 27.8 +/- 1.9 mg kg(-1) min(-1) (p < 0.05). Exogenous glucose utilisation rate during exercise was substantially greater (p < 0.001) in the HI study (18.4 +/- 2.1 mg kg(-1) min(-1)) than in the LO study (6.9 +/- 1.2 mg kg(-1) min(-1)). Muscle glycogen content fell by approximately 40% during exercise in both trials. Muscle glycogen utilisation, muscle intermediary metabolism, and phosphorylation of protein kinase B/Akt, glycogen synthase kinase 3alpha/beta and extracellular signal regulated protein kinase 1 and 2 proteins were no different between interventions. CONCLUSIONS/INTERPRETATION: In patients with type 1 diabetes, exercise under peak therapeutic insulin concentrations increases exogenous glucose utilisation but does not spare muscle glycogen utilisation. A disproportionate increase in exogenous glucose utilisation relative to the increase in CHO oxidation suggests an increase in glucose flux through non oxidative pathways. PMID- 17119917 TI - Peroxisome proliferator-activated receptor-delta, a regulator of oxidative capacity, fuel switching and cholesterol transport. AB - Synthetic agonists of peroxisome proliferator-activated receptor (PPAR)-delta have shown a promising pharmacological profile in preclinical models of metabolic and cardiovascular disease. At present, the pharmaceutical development of these drugs exploits the potential to raise plasma HDL-cholesterol in animals and their insulin-sensitising and glucose-lowering properties. PPAR-delta agonists have also proven to be powerful research tools that have provided insights into the role of fatty acid metabolism in human physiology and disease. Activation of PPAR delta induces the expression of genes important for cellular fatty acid combustion and an associated increase in whole-body lipid dissipation. The predominant target tissue in this regard is skeletal muscle, in which PPAR-delta activation regulates the oxidative capacity of the mitochondrial apparatus, switches fuel preference from glucose to fatty acids, and reduces triacylglycerol storage. These changes counter the characteristic derangements of insulin- resistant skeletal muscle but resemble the metabolic adaptation to regular physical exercise. Apart from effects on fuel turnover, there is evidence for direct antiatherogenic properties, because PPAR-delta activation increases cholesterol export and represses inflammatory gene expression in macrophages and atherosclerotic lesions. Whereas conclusions about the full potential of PPAR delta as a drug target await the result of large scale clinical testing, ongoing investigation of this nuclear receptor has greatly improved our knowledge of the physiological regulation of whole-body fuel turnover and the interdependence of mitochondrial function and insulin sensitivity. PMID- 17119918 TI - Meta-analysis of the effects of n-3 polyunsaturated fatty acids on haematological and thrombogenic factors in type 2 diabetes. AB - AIM/HYPOTHESIS: To determine whether marine-derived n-3 polyunsaturated fatty acids (n-3 PUFA) (also known as omega-3 fatty acids) have beneficial effects on haematological and thrombogenic risk markers in addition to dyslipidaemia, in patients with type 2 diabetes. METHODS: A systematic review and meta-analysis of randomised controlled trials comparing dietary or non-dietary intake of n-3 PUFA with placebo in type 2 diabetes was conducted by systematically searching databases from 1966 to February 2006. Changes in C-reactive protein, IL-6, TNF alpha, platelet function, fibrinogen, factor VII, von Willebrand factor, endothelial function, heart rate and blood pressure were recorded. Inclusion of studies, data extraction and quality were assessed independently in duplicate. RESULTS: Twelve trials involving 847 subjects with a mean treatment duration of 8.5 weeks included sufficient data to permit pooling. Compared with placebo, n-3 PUFA supplementation had a significant effect on two outcomes: reducing the level of diastolic blood pressure (five trials, 248 subjects) by a mean of 1.8 mm Hg (95% CI 0.0-3.6, p = 0.05) and increasing factor VII (two trials, 116 subjects) by 24.9% (95% CI 7.2-42.6, p = 0.006). There were no significant effects on systolic blood pressure, fibrinogen or heart rate. CONCLUSIONS/INTERPRETATION: These results suggest that, in addition to the recognised effects on dyslipidaemia, n-3 PUFA decreases diastolic blood pressure, and appears to increase factor VII. Larger and more rigorously conducted clinical trials are required to establish conclusively the role of n-3 PUFA in cardiovascular risk markers and clinical outcomes in type 2 diabetes. PMID- 17119919 TI - Beta(2)-Adrenergic activation increases glycogen synthesis in L6 skeletal muscle cells through a signalling pathway independent of cyclic AMP. AB - AIMS/HYPOTHESIS: In skeletal muscle, the storage of glycogen by insulin is regulated by glycogen synthase, which is regulated by glycogen synthase kinase 3 (GSK3). Here we examined whether adrenergic receptor activation, which can increase glucose uptake, regulates glycogen synthesis in L6 skeletal muscle cells. METHODS: We used L6 cells and measured glycogen synthesis (as incorporation of D: -[U-(14)C]glucose into glycogen) and GSK3 phosphorylation following adrenergic activation. RESULTS: Insulin (negative logarithm of median effective concentration [pEC(50)] 8.2 +/- 0.3) and the beta-adrenergic agonist isoprenaline (pEC(50) 7.5 +/- 0.3) induced a twofold increase in glycogen synthesis in a concentration-dependent manner. The alpha(1)-adrenergic agonist cirazoline and alpha(2)-adrenergic agonist clonidine had no effect. Both insulin and isoprenaline phosphorylated GSK3. The beta-adrenergic effect on glycogen synthesis is mediated by beta(2)-adrenoceptors and not beta(1)-/beta(3) adrenoceptors, and was not mimicked by 8-bromo-cyclic AMP or cholera toxin, and also was insensitive to pertussis toxin, indicating no involvement of cyclic AMP or inhibitory G-protein (G(i)) signalling in the beta(2)-adrenergic effect on glycogen synthesis. 12-O-tetra-decanoylphorbol-13-acetate (TPA) increased glycogen synthesis 2.5-fold and phosphorylated GSK3 fourfold. Inhibition of protein kinase C (PKC) isoforms with 12-(2-cyanoethyl)-6,7,12,13-tetrahydro-13 methyl-5-oxo-5H-indolo(2,3-a)pyrrollo(3,4-c)-carbazole (Go6976; inhibits conventional and novel PKCs) or 2-[1-(3-dimethylaminopropyl)-5-methoxyindol-3-yl] 3-(1H-indol-3-yl)maleimide (Go6983; inhibits conventional, novel and atypical PKCs) inhibited the stimulatory TPA effect, but did not significantly inhibit glycogen synthesis mediated by insulin or isoprenaline. Inhibition of phosphatidylinositol 3-kinase (PI3K) with wortmannin inhibited the effects of insulin and isoprenaline on glycogen synthesis. CONCLUSIONS/INTERPRETATION: These results demonstrate that in L6 skeletal muscle cells adrenergic stimulation through beta(2)-adrenoceptors, but not involving cyclic AMP or G(i), activates a PI3K pathway that stimulates glycogen synthesis through GSK3. PMID- 17119920 TI - Posterior reversible encephalopathy syndrome in intensive care medicine. AB - BACKGROUND: Posterior reversible encephalopathy syndrome (PRES) is a well recognized clinico-neuroradiological transient condition. Early recognition is of paramount importance for prompt control of blood pressure or removal of precipitating factors and treatment of epileptic seizures or status epilepticus. Delay in the diagnosis and treatment may in fact results in death or in irreversible neurological sequelae. DISCUSSION: PRES is characterized by headache, altered mental status, seizures, and visual disturbances and is associated with a number of different causes, most commonly acute hypertension, preeclampsia/eclampsia, and immunosuppressive agents. Clinical symptoms and neuroradiological findings are typically indistinguishable among the cases of PRES, regardless of underlying cause. Magnetic resonance studies typically show edema involving the white matter of cerebral posterior regions, especially parieto-occipital lobes but frontal and temporal lobes, and other encephalic structures may be involved. CONCLUSIONS: Intensivists and other physicians involved in the evaluation of patients with presumed PRES must be aware of the clinical spectrum of the associated conditions, the diagnostic modalities, and the correct treatment. PMID- 17119921 TI - Plasma volume expansion of 5% albumin, 4% gelatin, 6% HES 130/0.4, and normal saline under increased microvascular permeability in the rat. AB - OBJECTIVE: To compare the colloids 5% albumin, 4% gelatin, and 6% HES 130/0.4 with one another and with normal saline regarding their plasma expanding effects at increased permeability and to compare the results with those from a previous study at normal permeability. DESIGN AND SETTING: Prospective controlled randomized laboratory study in a university research laboratory. SUBJECTS: 48 adult male Sprague-Dawley rats. INTERVENTIONS: Permeability was increased by an injection of 0.5 ml dextran 70 using the fact that dextran causes anaphylactic reaction in the rat. Plasma volume was determined ((125)I albumin tracer technique) after anesthesia, 1 h after dextran injection (before infusion for 10 15 min of 20 ml/kg bw of each of the colloids or 80 ml/kg saline), and 3 h later. Blood pressure, hematocrit, blood gases, and electrolytes were measured. CVP was measured in four rats. MEASUREMENTS AND RESULTS: Plasma volume was 41.1+/-1.9 ml/kg at baseline (n=9), and 29.1+/-4.1 ml/kg (n=35) 1 h after the dextran injection. Three hours after infusion of the plasma expander plasma volume had increased by 17.1+/-3.4 ml/kg in the albumin group, 7.9+/-3.6 ml/kg in the gelatin group, 7.4+/-4.4 ml/kg in the HES group, and 12.2+/-3.1 ml/kg in the saline group. It was unchanged in a control group given no solution (n=7 for all groups). CONCLUSION: Albumin was a more effective plasma volume expander than gelatin or HES or saline (saline in 4 times larger volume). Gelatin and HES were equally effective. All solutions showed a smaller plasma expanding effect than observed in a previous study with normal permeability. PMID- 17119922 TI - Volume-targeted modes of modern neonatal ventilators: how stable is the delivered tidal volume? AB - OBJECTIVE: Volume-targeted modes are designed to deliver a constant tidal volume (V(t)) at lowest possible pressure independently of changes in compliance, resistance, and leak of the respiratory system. We examined whether these volume targeted modes respond rapidly enough to sudden changes in respiratory mechanics (e.g., selective intubation, surfactant administration, endotracheal tube kinking, de-kinking, obstruction), resulting in insufficient or excessive V(t) delivery. DESIGN AND SETTING: Bench study of six neonatal ventilators in the volume-targeted mode simulating preterm and full-term infant settings on a test lung. MEASUREMENTS AND RESULTS: Breath-to-breath expiratory V(t) were measured after rapid compliance, resistance, and leak changes. Under our test settings all ventilators showed important volume overshooting following rapid increase in compliance or decrease in resistance. Between one and 16 inflations were required to return to the set V(t). Some ventilators delivered inaccurate V(t) under steady state condition while others showed considerable breath-to-breath V(t) variability. CONCLUSIONS: We observed inaccurate V(t) delivery under specific conditions as well as immediate and sometimes prolonged volume overshooting after a rapid respiratory system compliance increase or resistance decrease in volume targeted modes of modern neonatal ventilators. Similar discrepancies between the set V(t) and the delivered inflations can be harmful in clinical situations, especially in newborns. Their clinical relevance needs to be clarified with safety studies in the neonatal population and we encourage manufacturers to further improve the ventilators algorithms. PMID- 17119923 TI - Goal-directed fluid management reduces vasopressor and catecholamine use in cardiac surgery patients. AB - OBJECTIVE: We examined whether guiding therapy by an algorithm based on optimizing the global end-diastolic volume index (GEDVI) reduces the need for vasopressor and inotropic support and helps to shorten ICU stay in cardiac surgery patients. DESIGN AND SETTING: Single-center clinical study with a historical control group at an university hospital. PATIENTS: Forty cardiac bypass surgery patients were included prospectively and compared with a control group. INTERVENTIONS: In the goal-directed therapy (GDT) group hemodynamic management was guided by an algorithm based on GEDVI. Hemodynamic goals were: GEDVI above 640 ml/m2, cardiac index above 2.5 l/min/m2, and mean arterial pressure above 70 mmHg. The control group was treated at the discretion of the attending physician based on central venous pressure, mean arterial pressure, and clinical evaluation. RESULTS: In the GDT group duration of catecholamine and vasopressor dependence was shorter (187+/-70 vs. 1458+/-197 min), and fewer vasopressors (0.73+/-0.32 vs. 6.67+/-1.21 mg) and catecholamines (0.01+/-0.01 vs. 0.83+/-0.27mg) were administered. They received more colloids (6918+/-242 vs. 5514+/-171ml). Duration of mechanical ventilation (12.6+/-3.6 vs. 15.4+/4.3 h) and time until achieving status of fit for ICU discharge (25+/-13 vs. 33+/-17h) was shorter in the GDT group. CONCLUSIONS: Guiding therapy by an algorithm based on GEDVI leads to a shortened and reduced need for vasopressors, catecholamines, mechanical ventilation, and ICU therapy in patients undergoing cardiac surgery. PMID- 17119924 TI - The "paraglider-wing" sign: an arthroscopic indicator of partial-thickness bursal surface tears of the supraspinatus tendon. AB - Partial-thickness bursal-surface tears of supraspinatus tendon may be missed on preoperative investigations and can be overlooked at surgery if not specifically sought. The authors describe an arthroscopic sign to detect these tears, when they involve more than half the tendon fibres, from the articular-side of the joint. The "paraglider-wing" sign, visualized during diagnostic glenohumeral arthroscopy, is demonstrated as an upward bulge of the capsulo-tendinous layer through the bursal-surface tear, under pressure of the inflow fluid. A positive sign indicates (1) presence of a partial-thickness bursal-side tear of the supraspinatus tendon, (2) significant depth (stage II or III) of the tear, and (3) the medial extent of the tear along the length of the tendon. A meticulous subacromial bursoscopy and excision of the bursa is then performed to visualize the tear from the subacromial space. Repair of the tear is performed with a double-row suture anchor fixation technique; the medial row of sutures is passed through the intact region of the tendon using the "paraglider-wing" sign as a guide. PMID- 17119925 TI - Carbon black particles increase reactive oxygen species formation in rat alveolar macrophages in vitro. AB - Alveolar macrophages (AM) have an important role in clearing particles from the lungs. In response to different stimuli they can release reactive oxygen species (ROS) and inflammatory mediators and promote pulmonary inflammation. We exposed rat AM to carbon black (CB) particles (0.63-20 microg/ml) and measured the eneration of ROS by using the fluorescent probe 2',7'-dichlorofluorescein diacetate. Fluorescence was elevated in a concentration dependent manner in the AM exposed to CB. Follow-up experiments using a series of enzyme inhibitors indicate that the ERK MAP kinase pathway and the p38 MAP kinase pathway may be involved in the formation of ROS. PMID- 17119926 TI - Effect of quinupristin/dalfopristin on 3T3 and Eahy926 cells in vitro in comparison to other antimicrobial agents with the potential to induce infusion phlebitis. AB - Infusion phlebitis is a common clinical problem that is observed with some antimicrobial agents, when being administered intravenously. In this study, cultured murine fibroblasts and immortalised human endothelial cells were exposed to three antibiotics at clinically relevant concentrations to assess their toxic potential in two established cytotoxicity assays. BALB/c 3T3 fibroblasts and Eahy926 endothelial cells were exposed to quinupristin/dalfopristin (QD), erythromycin and levofloxacin at increasing concentrations. For assessment of cytotoxicity the cells were incubated with neutral red (NR) or stained with crystal violet (CV). Measurements were done by photometry. At the concentration range tested QD and erythromycin showed a concentration-dependent cytotoxic effect in both cell cultures. In 3T3 cells the half-maximal effect concentration (EC50) was 20 mg/l for QD and 340 mg/l for erythromycin in the NR uptake test and 12 and 200 mg/l, respectively, in the CV assay. In Eahy926 cells the EC50 was 50 mg/l for QD and 880 mg/l for erythromycin in the NR uptake test and 40 and 750 mg/l, respectively, in the CV assay. No EC50 could be established in both cell types for levofloxacin. Eahy926 cells were less sensitive to cytotoxic stimuli than 3T3 fibroblasts. Cytotoxic effects in both cell cultures occurred in the following order: QD > erythromycin >> levofloxacin. This ranking correlates well with the frequency of local adverse effects observed with the infusion of these antibiotics in patients. Thus, these in vitro assays may serve as an estimate for the prediction of local tolerability of antibiotics when administered parenterally. PMID- 17119927 TI - Open channel block of Kv1.3 by rosiglitazone and troglitazone: Kv1.3 as the pharmacological target for rosiglitazone. AB - The effects of rosiglitazone and troglitazone were examined on cloned Kv1.3 channels stably expressed in Chinese hamster ovary cells using the whole-cell configuration of the patch-clamp technique. Rosiglitazone decreased the Kv1.3 currents and accelerated the decay rate of current inactivation in a concentration-dependent manner with an IC(50) of 18.6 microM. These effects were reversible after washout of the drug. Troglitazone caused the block of Kv1.3 with a similar pattern but was five times more potent than rosiglitazone with an IC(50) of 3.5 microM. The block of Kv1.3 by rosiglitazone and troglitazone was voltage-dependent at a membrane potential coinciding with the activation of the channels. Both drugs decreased the tail current amplitude and slowed the deactivation process of Kv1.3, resulting in a tail crossover phenomenon. These results indicate that rosiglitazone and troglitazone block the open state of Kv1.3 channels, suggesting that it is an important pharmacological target for rosiglitazone as a potent blocker of Kv1.3 channels. PMID- 17119928 TI - CNS drugs approved by the centralised European procedure: true innovation or dangerous stagnation? PMID- 17119929 TI - Calculating utility: preclinical evidence for cost-benefit analysis by mesolimbic dopamine. AB - RATIONALE: Throughout our lives we constantly assess the costs and benefits of the possible future outcomes of our actions and use this information to guide behavior. There is accumulating evidence that dopamine contributes to a fundamental component of this computation-how rewards are compared with the costs incurred when obtaining them. OBJECTIVE: We review the evidence for dopamine's role in cost-benefit decision making and outline a simple mathematical framework in which to represent the interactions between rewards, costs, behavioral state and dopamine. CONCLUSIONS: Dopamine's effects on cost-benefit decision making can be modeled using simple utility-function curves. This approach provides a useful framework for modeling existing data and generating experimental hypotheses that can be objectively and quantitatively tested by observing choice behavior without the necessity to account for subjective psychological states such as pleasure or desire. We suggest that dopamine plays a key role in overcoming response costs and enabling high-effort behaviors. A particularly important anatomical site of this action is the core of the nucleus accumbens. Here, dopamine is able to modulate activity originating from the frontal cortical systems that also assess costs and rewards. Internal deprivation states (e.g., hunger and thirst) also help to energize goal-seeking behaviors, probably in part by their rich influence on dopamine, which can in turn modify decision making policies. PMID- 17119930 TI - CP-154,526, a CRF type-1 receptor antagonist, attenuates the cue-and methamphetamine-induced reinstatement of extinguished methamphetamine-seeking behavior in rats. AB - RATIONALE: Previous studies from our laboratory and others have indicated a role for the hypothalamo-pituitary-adrenal (HPA) axis in the extinction/reinstatement animal model of cocaine relapse OBJECTIVE: This present study was designed to investigate the potential role for the HPA axis in the cue- and methamphetamine induced reinstatement of extinguished methamphetamine-seeking behavior by determining the effects of ketoconazole and the corticotropin-releasing hormone (CRF) type 1 receptor antagonist, CP-154,526, on these behaviors. MATERIALS AND METHODS: Male Wistar rats were trained to self-administer methamphetamine (0.03 mg/kg/infusion). The delivery of methamphetamine was paired with the presentation of a tone and the illumination of a house light. Once stable responding was reached, the rats were placed into extinction. The effects of pretreatment with ketoconazole (25, 50, or 100 mg/kg, i.p.) or CP-154,526 (20 or 40 mg/kg, i.p.; 3 micro g, i.c.v) on cue-induced reinstatement were then evaluated. RESULTS: Cue induced reinstatement was not significantly attenuated by pretreatment with peripherally administered CP-154,526 or by pretreatment with ketoconazole. However, centrally administered CP-154,526 (3 micro g, i.c.v.) significantly attenuated cue-induced reinstatement. In a separate group of rats, CP-154,526 (20 mg/kg, i.p.) attenuated methamphetamine-induced reinstatement (0.12 mg/kg priming infusion); whereas a higher dose (40 mg/kg) was necessary to attenuate reinstatement induced by a priming infusion of 0.24 mg/kg/infusion. Ketoconazole (50 mg/kg) did not affect reinstatement induced by a 0.12 mg/kg priming infusion and, therefore, was not tested at the higher methamphetamine priming dose. CONCLUSIONS: These data suggest an important role for CRF in the cue- and methamphetamine-induced reinstatement of extinguished methamphetamine-seeking behavior. PMID- 17119931 TI - Pharmacokinetic and behavioral characterization of a long-term antipsychotic delivery system in rodents and rabbits. AB - RATIONALE: Non-adherence with medication remains the major correctable cause of poor outcome in schizophrenia. However, few treatments have addressed this major determinant of outcome with novel long-term delivery systems. OBJECTIVES: The aim of this study was to provide biological proof of concept for a long-term implantable antipsychotic delivery system in rodents and rabbits. MATERIALS AND METHODS: Implantable formulations of haloperidol were created using biodegradable polymers. Implants were characterized for in vitro release and in vivo behavior using prepulse inhibition of startle in rats and mice, as well as pharmacokinetics in rabbits. RESULTS: Behavioral measures demonstrate the effectiveness of haloperidol implants delivering 1 mg/kg in mice and 0.6 mg/kg in rats to block amphetamine (10 mg/kg) in mice or apomorphine (0.5 mg/kg) in rats. Additionally, we demonstrate the pattern of release from single polymer implants for 1 year in rabbits. CONCLUSIONS: The current study suggests that implantable formulations are a viable approach to providing long-term delivery of antipsychotic medications in vivo using animal models of behavior and pharmacokinetics. In contrast to depot formulations, implantable formulations could last 6 months or longer. Additionally, implants can be removed throughout the delivery interval, offering a degree of reversibility not available with depot formulations. PMID- 17119933 TI - Simultaneous determination of losartan, EXP-3174 and hydrochlorothiazide in plasma via fully automated 96-well-format-based solid-phase extraction and liquid chromatography-negative electrospray tandem mass spectrometry. AB - An automated, sensitive and high-throughput liquid chromatographic/electrospray tandem mass spectrometric (LC-MS/MS) assay was developed for the simultaneous determination of losartan (LOS), its major circulating metabolite EXP-3174 and hydrochlorothiazide (HCTZ) in human plasma. LOS and HCTZ coexist in the same drug formulation, and this is the first method that enables the simultaneous determination of both drugs along with the active metabolite of LOS. Since these drugs have different physicochemical properties, the employment of a liquid liquid extraction (LLE) protocol was precluded. A fully automated solid-phase extraction (SPE) protocol, based on 96-well format plates, was used to isolate these compounds and furosemide (internal standard, IS) from plasma. Washing and elution steps were amended accordingly in order to minimize any matrix effect from components of the plasma without reducing the elution of the molecules of interest. The compounds were eluted from a C18 column and detected with an API 3000 triple-quadrupole mass spectrometer using negative electrospray ionization and multiple reaction monitoring (MRM). The assay was linear over the range 1.00 400 ng/mL for LOS and EXP-3174 and 0.500-200 ng/mL for HCTZ, respectively, when 200 microl of plasma was used in the extraction. The overall intra- and interassay variations were within acceptance limits. The analysis time for each sample was 4 min, and more than 300 samples could be analyzed in one day by running the system overnight. The assay was simple, highly sensitive, selective, precise, fast, and it enables the reliable determination of LOS, EXP-3174 and HCTZ in pharmacokinetic or bioequivalence studies after per os administration of a single tablet containing both drugs. PMID- 17119934 TI - Biosensor incorporating cell barrier architectures for detecting Staphylococcus aureus alpha toxin. AB - Alpha toxin is a common virulent factor of Staphylococcus aureus and is believed to play crucial roles in pathogenicity induced by S. aureus. Alpha toxin is also known to induce permeability to endothelial cell monolayers in vitro due to the formation of interendothelial gaps. The present study is directed towards measuring alpha toxin using a whole-cell-based biosensor. The biosensor, consisting of a confluent monolayer of human umbilical vein endothelial cells (HUVECs) on a potassium ion-selective electrode, takes advantage of cell permeability dysfunction to detect the presence of small quantities of alpha toxin. When a confluent monolayer of cells was formed on the membrane surface, the response of the electrode toward the marker ion, potassium, was inhibited. Upon exposing this sensor to varying concentrations of alpha toxin for 20 min, an increase in sensor response to potassium was observed. The response thus obtained was indirectly related to the concentration of alpha toxin. The detection limit of this sensor for alpha toxin was found to be 0.1 ng/ml. Cell monolayers were stained with silver nitrate to quantify the formation of intercellular gaps as well as to study the effect of this toxin on HUVECs morphology. A strong positive correlation was observed between the response obtained from the biosensor and the area of the intercellular gaps. Silver staining also revealed the tendency of cells to round up upon being exposed to alpha toxin. PMID- 17119935 TI - Determination of NSAIDs in river sediment samples. AB - Consumption of non-steroidal anti-inflammatory drugs (NSAIDs) is increasing and with it the danger of environmental pollution by pharmaceutical residues. Publications regarding NSAIDs in the environment not only show that they are toxic to many animal species, but also highlight the need for robust analytical methods for monitoring the level of such contaminants in environmental matrices. In our study we selected the four most widely used NSAIDs in Slovenia and Central Europe, ibuprofen, naproxen, ketoprofen and diclofenac, and studied their extraction from sediment samples. We examined several extraction techniques (ultrasonic extraction, Soxhlet extraction, pressurized liquid extraction, supercritical fluid extraction and microwave-assisted extraction) using a spiked sediment sample and determined optimal extraction conditions. After extraction we applied a clean-up step, derivatisation of the analytes and gas chromatography with mass spectrometric detection (GC-MSD) and selected the most appropriate extraction procedure. The optimised analytical method chosen for analysis of sediment samples consisted of microwave-assisted extraction, clean-up of the extract with SPE, derivatisation with MSTFA and determination with GC-MSD. The optimised procedure was applied to the analysis of two environmental river samples taken from the vicinity of Novo mesto, the biggest town in the south eastern part of Slovenia with 62,000 inhabitants, a hospital and a pharmaceutical factory in its vicinity. While analysis of the sample taken upstream of the town showed no detectable amounts of NSAIDs, analysis of samples taken downstream showed quantifiable levels of two of the studied NSAIDs (naproxen and ketoprofen). Besides these two NSAIDs, river water samples sampled at the same time and location on the River Krka also showed the presence of diclofenac. Sampling on the River Krka and other Slovene rivers will in the future be repeated at different sampling points in order to track down the main sources of pollution. PMID- 17119936 TI - Microanalyzer for biomonitoring lead (Pb) in blood and urine. AB - Biomonitoring of lead (Pb) in blood and urine enables quantitative evaluation of human occupational and environmental exposures to Pb. State-of-the-art ICP-MS instruments can only analyze metals in laboratories, resulting in lengthy turnaround times, and they are expensive. In response to the growing need for a metal analyzer capable of on-site, real-time monitoring of trace toxic metals in individuals, we developed a portable microanalyzer based on flow injection/stripping voltammetry (ASV), and validated the system using rat blood and urine spiked with known amounts of Pb. Fouling of electrodes by proteins often prevents the effective use of electrochemical sensors in biological matrices. Minimization of such fouling was accomplished with suitable sample pretreatment and by establishing turbulent flow of blood and urine containing Pb onto the electrode inside the microanalyzer, which resulted in no apparent electrode fouling even when the samples contained 50% urine or 10% blood by volume. No matrix effect was observed for the voltammetric Pb signals, even when the samples contained 10% blood or 10% urine. The microanalyzer offered linear concentration ranges relevant to Pb exposure levels in humans (0-20 ppb in 10% blood samples, 0-50 ppb in 50% urine samples). The device showed excellent sensitivity and reproducibility; Pb detection limits were 0.44 ppb and 0.46 ppb, and % R.S.D. was 4.9 and 2.4 in 50% urine and 10% blood samples, respectively. It gave similar Pb concentrations in blood and urine to those measured by ICP-MS. It offered high throughput (3 min per sample) and economical use of samples (60 microL per measurement) as well as low reagent consumption (1 microg of Hg per measurement), thus minimizing environmental concerns associated with mercury use. Since it is miniaturized, the microanalyzer is portable and field-deployable. Thus, it shows much promise as the next-generation analyzer for the biomonitoring of toxic metals. PMID- 17119937 TI - Development of a novel luminol chemiluminescent method catalyzed by gold nanoparticles for determination of estrogens. AB - It has been found that gold nanoparticles (nano-Au) enhance the chemiluminescence (CL) of the luminol-hydrogen peroxide system and that estrogens inhibit these CL signals in alkaline solution. CL spectra, UV-visible spectra, X-ray photoelectron spectra (XPS), and transmission electron microscopy (TEM) were used to investigate the mechanism of the CL enhancement. On the basis of the inhibition, a flow-injection CL method has been established for determination of three natural estrogens. Under the optimized conditions, the linear range for determination of the estrogens was 0.07 to 7.0 micromol L(-1) for estrone, 0.04 to 10 micromol L(-1) for estradiol, and 0.1 to 10 micromol L(-1) for estriol. The detection limits were 3.2 nmol L(-1) for estrone, 7.7 nmol L(-1) for estradiol, and 49 nmol L(-1) for estriol, with RSD of 2.9, 2.6, and 1.8%, respectively. This method has been used for analysis of estrogens in commercial tablets and in urine samples from pregnant women. PMID- 17119938 TI - Chemiluminescent detection of DNA hybridization using gold nanoparticles as labels. AB - A chemiluminescent (CL) detection method has been developed for DNA hybridization. The assay relies on a sandwich-type DNA hybridization in which gold nanoparticles modified with alkylthiol-capped oligonucleotide strands are used as probes to monitor the presence of the specific target DNA. The AuCl(4)( ), which is the dissolving product of the gold nanoparticles anchored on the DNA hybrids, serves as an analyte in the H(2)O(2)-luminol- AuCl(4)(-) CL reaction for the indirect measurement of the target DNA. The combination of the remarkable sensitivity of the CL analysis with the large number of AuCl(4)(-) released from each DNA hybrid allows a detection limit at levels as low as 0.1 pM of the target DNA. Moreover, with a further silver amplification step, the detection limit will be pushed down to the femtomolar domain. PMID- 17119939 TI - Rhythmic arm cycling produces a non-specific signal that suppresses Soleus H reflex amplitude in stationary legs. AB - Rhythmic arm cycling significantly suppresses Hoffmann (H-) reflex amplitude in Soleus muscles of stationary legs. The specific parameters of arm cycling contributing to this suppression, however, are unknown. Between the arms or legs, movement results in suppression of the H-reflex that is specifically related to the phase of movement and the locus of limb movement. We speculated that the effects of arm movement features on H-reflexes in the leg would be similar and hypothesized that the Soleus H-reflex suppression evoked by arm movement would therefore be specifically related to: (1) phase of the movement; (2) the locus of the movement (i.e., ipsilateral or contralateral arm); (3) range of arm motion; and (4) frequency of arm cycling. Participants performed bilateral arm cycling at 1 and 2 Hz with short and long-crank lengths. Ipsilateral and contralateral arm cycling was also performed at 1 Hz with a long-crank length. Soleus H-reflexes were evoked at four equidistant phases and comparisons were made while maintaining similar evoked motor waves and Soleus activation. Our results show that comparable suppressive effects were seen at all phases of the arm movement: there was no phase-dependence. Further, bilateral or unilateral (whether ipsi- or contralateral arm) cycling yielded equivalent suppression of the H-reflex amplitude. Cycling at 2 Hz resulted in a significantly larger suppression than with 1 Hz cycling. We conclude that a general, rather than a specific, signal related to the command to produce rhythmic arm muscle activity mediates the suppression of Soleus H-reflex during arm cycling. PMID- 17119940 TI - The time course of visuo-motor affordances. AB - To measure dynamic visuo-motor coupling within the two hemispheres, we showed observers an animation of a rotating cup. They indicated with left or right buttons when a fixation dot in the center of the animation changed color. For either hand, response times changed continuously with the irrelevant position of the cup's handle, even when the perceptual asymmetry of the handle was controlled. This spontaneous lateralized motor preparation showed both top-down and bottom-up components and was more pronounced for the right than the left hand. The dynamic affordance method will help understand perception-action coupling in the brain. PMID- 17119941 TI - Returning home: location memory versus posture memory in object manipulation. AB - Previous studies of object manipulation have suggested that when participants return an object to the place from which they just carried it, they tend to grasp the object for the target-back-to-home trips close to where they just grasped it for the home-to-target trips [Exp Brain Res 157(4):486-495, 2004; Psychon Bull Rev, 2006]. What was unclear from these previous studies was whether participants recalled postures or locations. According to the posture hypothesis, they remembered what body positions they adopted when they last held the object. According to the location hypothesis, they remembered where they held the object and then took hold of it there or nearby again. To distinguish between these possibilities, we had participants mount or dismount a platform after home-to target moves and before target-back-to-home moves. In the control condition, they did not change their vertical position relative to the shelf containing the home and target platforms (they merely stepped sideways). We found that participants grasped the object at nearly the same place along its length as they had before, even if this meant adopting very different postures than before. This outcome is consistent with the location-recall account and is inconsistent with the posture recall account. The implications for motor planning are discussed. PMID- 17119942 TI - Dual adaptation to two opposing visuomotor rotations when each is associated with different regions of workspace. AB - Studies examining dual adaptation to opposing novel environments have yielded contradictory results, with previous evidence supporting both successful dual adaptation and interference leading to poorer adaptive performance. Whether or not interference is observed during dual adaptation appears to be dependent on the method used to allow the performer of the task to distinguish between two novel environments. This experiment tested if colour cues, a separation in workspace, and presentation schedule, could be used to distinguish between two opposing visuomotor rotations and enable dual adaptation. Through the use of a purpose designed manipulandum, each visuomotor rotation was either presented in the same region of workspace and associated with colour cues (Group 1), different regions of workspace in addition to colour cues (Groups 2 and 3) or different regions of workspace only (Groups 4 and 5). We also assessed the effectiveness of the workspace separation with both randomised and alternating presentation schedules (Groups 4 and 5). The results indicated that colour cues were not effective at enabling dual adaptation when each of the visuomotor rotations was associated with the same region of workspace. When associated with different regions of workspace, however, dual adaptation to the opposing rotations was successful regardless of whether colour cues were present or the type of presentation schedule. PMID- 17119943 TI - Interactions between luminance and colour channels in visual search and their relationship to parallel neural channels in vision. AB - We have compared visual search under conditions that tend to isolate the magnocellular, parvocellular and koniocellular channels of the human visual system. We used isoluminant red-green stimuli that do not modulate short wavelength sensitive (SWS) cones to isolate the parvocellular pathway, isoluminant SWS-cone isolating stimuli to stimulate only the koniocellular system and addition of small luminance contrasts to selectively activate the magnocellular pathway. We found that in the case of conjunction search, where attentional resources were required, the red-green (parvocellular) system can use accompanying small luminance (magnocellular) signals to improve visual search. On the other hand, when using SWS-cone isolating stimuli to selectively stimulate the blue-yellow (koniocellular) system, addition of similar luminance signals did not increase the efficiency of the serial visual search. The results indicate that S-cone signals may be processed in a separate pathway that does not get converging inputs from the magnocellular pathway. This is unlike the case with the red-green opponent system, which functions more synergistically with the magnocellular system. PMID- 17119944 TI - Modification of CYP2E1 and CYP3A4 activities in haemoglobin E-beta thalassemia patients. AB - OBJECTIVE: Thalassemia disease is a genetic haemoglobinopathy usually associated with an iron overload and some degree of organ impairment. The impact of the disease on the drug metabolising enzyme cytochrome P450 (CYP) is not known. CYP2E1 and CYP3A4 are responsible for the metabolism of a large number of drugs and changes in their activities may have clinical consequences. METHODS: Haemoglobin E-beta thalassemia paediatric, blood transfusion-dependent patients apparently without complications (n = 35) and healthy controls (n = 42) were recruited in this study. The ratios of plasma 6-hydroxychlorzoxazone to chlorzoxazone, and urinary 6-beta-hydroxycortisol (6beta-OHF) to cortisol were used as indices for CYP2E1 and CYP3A4 activities, respectively. Blood and plasma samples were assayed for parameters of clinical biochemistry, oxidants and antioxidants. RESULTS: There were significant increases in serum iron, protein carbonyl and lipid peroxidation in thalassemia patients, whereas there was a decrease in blood glutathione, but unchanged plasma nitric oxide metabolites. CYP2E1 activity in the patients was unchanged; however, when the patients were stratified by splenectomy status, CYP2E1 activity was increased in non splenectomised patients in comparison with the controls and splenectomised subjects. On the other hand, 6beta-OHF/cortisol ratios increased markedly in patients associated with depressed growth hormone levels. There were no correlations between CYP2E1 activity and oxidant stress or antioxidant parameters. CONCLUSIONS: This report is the first demonstration that thalassemia major is associated with an alteration of CYP2E1 and CYP3A4 activities; this could modify the sensitivity of thalassemia patients to the toxic or therapeutic effects of drugs. PMID- 17119945 TI - Hepatic adverse drug reactions: a case/non-case study in Italy. AB - OBJECTIVE: Adverse drug reactions (ADRs) can involve all tissues and organs. Liver injuries are considered among the most serious and are a cause for concern among physicians and patients. To assess the extent of drug-induced liver injuries in Italy we compared the number of cases of hepatic ADRs with reports of all other drug-related reactions present in the same database. METHODS: Spontaneous reports from six Italian Regions collected from January 1990 to May 2005 were analysed. Adverse reactions were classified according to WHO Adverse Reaction Terminology for causality assessment, and only those with "certain", "probable" or "possible" causality assessment were included. Association between drugs and hepatic ADRs was assessed using the case/non case method, calculating the ADR reporting odds ratio (ROR) as a measure of disproportionality. RESULTS: On May 2005, the database contained 35,767 ADR reports, of which 11,829 were excluded because they were unclassifiable or unlikely in terms of causality assessment. Therefore, the analysis was carried out on 23,938 reports, of which 1,069 concerned hepatic ADRs (cases) and 22,869 concerned non-cases. The proportion of serious ADRs was about 40% in the overall database, and about 74% among cases. The drug classes with the highest number of cases were statins (ROR = 2.9, 95% CI 2.4-3.5), antiplatelet agents (ROR = 3.5; 95% CI 2.6-4.6), NSAIDs (ROR = 2.9; 95% CI 2.1-3.9) and macrolides (ROR = 1.7; 95% CI 1.2-2.3). CONCLUSION: Hepatic adverse drug reactions remain a serious concern for several drugs widely used in clinical practice. Monitoring hepatic enzymes on a monthly basis for the first 6 months of treatment has been suggested for patients taking medications known to be hepatotoxic. A better knowledge of the epidemiology and mechanisms of hepatic ADRs may contribute to minimising their occurrence. PMID- 17119946 TI - Magnetic resonance imaging assessment of brain maturation in preterm neonates with punctate white matter lesions. AB - INTRODUCTION: Early white matter (WM) injury affects brain maturation in preterm infants as revealed by diffusion tensor imaging and volumetric magnetic resonance (MR) imaging at term postmenstrual age (PMA). The aim of the study was to assess quantitatively brain maturation in preterm infants with and without milder forms of WM damage (punctate WM lesions, PWML) using conventional MRI. METHODS: Brain development was quantitatively assessed using a previously validated scoring system (total maturation score, TMS) which utilizes four parameters (progressive myelination and cortical infolding, progressive involution of glial cell migration bands and germinal matrix tissue). PWML were defined as foci of increased signal on T1-weighted images and decreased signal on T2-weighted images with no evidence of cystic degeneration. A group of 22 preterm infants with PWML at term PMA (PWML group) were compared with 22 matched controls with a normal MR appearance. RESULTS: The two groups were comparable concerning gestational age, birth weight and PMA. TMS was significantly lower in the PWML group than in the control group (mean TMS 12.44 +/- 2.31 vs 14.00 +/- 1.44; P = 0.011). Myelination (mean 2.76 +/- 0.42 PWML group vs 3.32 +/- 0.55 control group, P = 0.003) and cortical folding (3.64 +/- 0.79 vs 4.09 +/- 0.43, P = 0.027) appeared to be significantly delayed in babies with PWML. CONCLUSION: Conventional MRI appears able to quantify morphological changes in brain maturation of preterm babies with PWML; delayed myelination and reduced cortical infolding seem to be the most significant aspects. PMID- 17119947 TI - HydroCoil as an adjuvant to bare platinum coil treatment of 100 cerebral aneurysms. AB - INTRODUCTION: The overall safety of the HydroCoil, an expansile hybrid hydrogel platinum coil, is unknown. We report a prospective observational study of our first 100 cerebral aneurysms treated with HydroCoils, focusing on safety and initial efficacy. METHODS: Indications, procedural complications, clinical and angiographic outcomes were recorded. Packing density, number of coils deployed and angiographic results were compared with those in a matched control group of 100 aneurysms treated solely with bare platinum coils. HydroCoil complication rates were compared to bare platinum coil rates at our institution and in published series. RESULTS: Adjuvant HydroCoil treatment led to increased mean percentage aneurysm filling compared to controls (50 +/- 21% versus 27 +/- 13%, P < 0.001). Immediate posttreatment angiographic results showed significantly (P < 0.001) more complete occlusions and fewer incomplete (<95%) occlusions compared to controls. Intermediate follow-up angiograms (median 7.5 months) in 63 aneurysms showed a trend towards fewer incomplete occlusions with HydroCoil treatment. There were significantly fewer major recurrences with HydroCoil treatment compared to the control treatment (9.5% versus 22.6%, P = 0.046). In the adjuvant HydroCoil group, major recurrent aneurysms had significantly less percentage volume packing with HydroCoils than non-recurrent aneurysms (50.3 +/- 5.0% versus 65.3 +/- 18.0%, P = 0.04). There was a 12% procedural complication rate, 6% procedural morbidity and 1% mortality rate, similar to institutional and reported bare platinum coil complication rates. CONCLUSION: HydroCoils can be safely deployed with a similar complication rate to bare platinum coils. They result in improved aneurysm filling. Intermediate follow-up angiography showed significantly fewer major recurrences. Long-term follow-up is required to confirm initial improved stability. PMID- 17119948 TI - Functional MRI of the cervical spinal cord on 1.5 T with fingertapping: to what extent is it feasible? AB - INTRODUCTION: Until recently, functional magnetic resonance imaging (fMRI) with blood oxygen level-dependent (BOLD) contrast, was mainly used to study brain physiology. The activation signal measured with fMRI is based upon the changes in the concentration of deoxyhaemoglobin that arise from an increase in blood flow in the vicinity of neuronal firing. Technical limitations have impeded such research in the human cervical spinal cord. The purpose of this investigation was to determine whether a reliable fMRI signal can be elicited from the cervical spinal cord during fingertapping, a complex motor activity. Furthermore, we wanted to determine whether the fMRI signal could be spatially localized to the particular neuroanatomical location specific for this task. METHODS: A group of 12 right-handed healthy volunteers performed the complex motor task of fingertapping with their right hand. T2*-weighted gradient-echo echo-planar imaging on a 1.5-T clinical unit was used to image the cervical spinal cord. Motion correction was applied. Cord activation was measured in the transverse imaging plane, between the spinal cord levels C5 and T1. RESULTS: In all subjects spinal cord responses were found, and in most of them on the left and the right side. The distribution of the activation response showed important variations between the subjects. While regions of activation were distributed throughout the spinal cord, concentrated activity was found at the anatomical location of expected motor innervation, namely nerve root C8, in 6 of the 12 subjects. CONCLUSION: fMRI of the human cervical spinal cord on an 1.5-T unit detects neuronal activity related to a complex motor task. The location of the neuronal activation (spinal cord segment C5 through T1 with a peak on C8) corresponds to the craniocaudal anatomical location of the neurons that activate the muscles in use. PMID- 17119949 TI - Incidental internal carotid artery calcifications on temporal bone CT in children. AB - BACKGROUND: Incidental internal carotid artery (ICA) calcifications are occasionally noted on CT images of the brain and temporal bone. In adults, incidental calcifications have been correlated with increased incidence of hypercholesterolemia, cardiac disease, diabetes and carotid stenosis. OBJECTIVE: To determine the incidence of incidental calcifications of the carotid siphon on temporal bone CT in children. MATERIALS AND METHODS: We retrospectively reviewed 24 months of consecutive temporal bone CT examinations in children aged 18 years and younger. CT examinations on 663 patients were reviewed and the presence or absence of ICA calcifications was ranked as absent, questionable or definitive. In patients in whom definitive calcifications were identified, hospital charts were reviewed for evidence of diabetes mellitus, hypercholesterolemia, hypertriglyceridemia, hyperlipidemia and chronic renal disease as potential causes of early atherosclerosis. RESULTS: Of the 663 patients, 25% had definitive calcifications within the wall of the ICA: 6% of children younger than 2 years and 28% of children 12-19 years of age. CONCLUSIONS: Incidentally noted ICA calcifications are a common finding on temporal bone CT in children, most likely a physiologic response to turbulent flow at natural bends in the artery rather than secondary to underlying disease predisposing to early atherosclerotic calcification. PMID- 17119950 TI - Two critical genes (HLA-DRB1 and ABCF1)in the HLA region are associated with the susceptibility to autoimmune pancreatitis. AB - We have previously reported that autoimmune pancreatitis (AIP) is a bioclinical entity characterized by high serum immunoglobulin G4 concentrations and association with the HLA-DRB1*0405-DQB1*0401 haplotype. However, the precise identity of gene(s) within this haplotype directly responsible for AIP pathogenesis is yet to be established. To dissect the genetic contribution of the incriminated haplotype, we have now performed an association analysis within the human leukocyte antigen (HLA) region using various types of polymorphic markers. Genomic DNAs from 43 AIP patients and 213 unrelated Japanese controls were used in this analysis. In each DNA sample, we established the genotype of 25 microsatellite markers distributed throughout the HLA region, that of single nucleotide polymorphism within the 5'-flanking regions of the TNFA and IkBLI (also known as NFKBIL1) as well as HLA class I and II genes. The HLA-linked susceptibility regions for AIP were localized to two segments: HLA-DRB1 (*0405; OR = 3.20, P = 0.00063, Pc = 0.0016) -DQB1 (*0401; OR = 3.29, P = 0.00046, Pc = 0.0069) in the HLA class II and C3-2-11 microsatellite (allele 219; OR = 2.96, P = 0.0076, Pc = 0.099) in the HLA class I regions. Upon stratification analysis in search for a synergistic effect given the extensive linkage disequilibrium within the major histocompatibility complex, it was established that each segment contributed to disease pathogenesis. The two critical HLA regions for susceptibility to AIP are limited to the HLA-DRB1*0405-DQB1*0401 in the class II and the ABCF1 proximal to C3-2-11, telomeric of HLA-E, in the class I regions. PMID- 17119951 TI - A modular concept of HLA for comprehensive peptide binding prediction. AB - A variety of algorithms have been successful in predicting human leukocyte antigen (HLA)-peptide binding for HLA variants for which plentiful experimental binding data exist. Although predicting binding for only the most common HLA variants may provide sufficient population coverage for vaccine design, successful prediction for as many HLA variants as possible is necessary to understand the immune response in transplantation and immunotherapy. However, the high cost of obtaining peptide binding data limits the acquisition of binding data. Therefore, a prediction algorithm, which applies the binding information from well-studied HLA variants to HLA variants, for which no peptide data exist, is necessary. To this end, a modular concept of class I HLA-peptide binding prediction was developed. Accurate predictions were made for several alleles without using experimental peptide binding data specific to those alleles. We include a comparison of module-based prediction and supertype-based prediction. The modular concept increased the number of predictable alleles from 15 to 75 of HLA-A and 12 to 36 of HLA-B proteins. Under the modular concept, binding data of certain HLA alleles can make prediction possible for numerous additional alleles. We report here a ranking of HLA alleles, which have been identified to be the most informative. Modular peptide binding prediction is freely available to researchers on the web at http://www.peptidecheck.org . PMID- 17119952 TI - Molecular characterization of coding sequences and analysis of Toll-like receptor 3 mRNA expression in water buffalo (Bubalus bubalis) and nilgai (Boselaphus tragocamelus). AB - Toll-like receptor 3 (TLR3), an antiviral innate immunity receptor recognizes double-stranded RNA, preferably of viral origin and induces type I interferon production, which causes maturation of phagocytes and subsequent release of chemical mediators from phagocytes against some viral infections. The present study has characterized TLR3 complementary DNA (cDNA) in buffalo (Bubalus bubalis) and nilgai (Boselaphus tragocamelus). TLR3 coding sequences of both buffalo and nilgai were amplified from cultured dendritic cell cDNA and cloned in pGEMT-easy vector for characterization by restriction endonucleases and nucleotide sequencing. Sequence analysis reveals that 2,715-bp-long TLR3 open reading frame encoding 904 amino acids in buffalo as well as nilgai is similar to that of cattle. Buffalo TLR3 has 98.6 and 97.9% identity at nucleotide level with nilgai and cattle, respectively. Likewise, buffalo TLR3 amino acids share 96.7% identity with cattle and 97.8% with nilgai. Non-synonymous substitutions exceeding synonymous substitutions indicate evolution of this receptor through positive selection among these three ruminant species. Buffalo and nilgai appear to have diverged from a common ancestor in phylogenetic analysis. Predicted protein structures of buffalo and nilgai TLR3 from deduced amino acid sequences indicate that the buffalo and nilgai TLR3 ectodomain may be more efficient in ligand binding than that of cattle. Furthermore, TLR3 messenger RNA expression in tissues as quantified by real-time PCR was found higher in nilgai than buffalo. PMID- 17119953 TI - Differential calcium signaling in dairy cows with specific CXCR1 genotypes potentially related to interleukin-8 receptor functionality. AB - Neutrophil migration and activation are critical components of innate immunity and are mediated by a variety of inflammatory mediators, which include interleukin-8 (IL-8) and epithelial-derived neutrophil activating peptide-78 (ENA 78). Limited knowledge on the expression of receptors for these inflammatory mediators (CXCR1 and CXCR2) in bovine, in addition to the association of a polymorphism (G-->C) in position +777 of the CXCR1 gene with impaired neutrophil function, prompted evaluation of CXCR1 and CXCR2 mRNA and protein expression, ligand binding affinity, and intracellular receptor signaling in neutrophils from cows with different CXCR1 genotypes. Initial observations revealed that overall IL-8 receptor numbers appeared to be lower in cows with a CC genotype compared to cows with a GG genotype. However, in the presence of SB225002, a CXCR2 inhibitor, CXCR1 affinity was about fivefold lower in cows with a CC genotype and may have resulted in an underestimation of receptor numbers in cows with this genotype. In addition, intracellular calcium ([Ca++]i) release was lower in cows with a CC genotype when cells were stimulated with IL-8 but not ENA-78. Furthermore, when neutrophils were stimulated with an optimal dose of IL-8 in the presence of SB225002, [Ca++]i release was lower in cows with a CC genotype, suggesting differential CXCR1 signaling among genotypes. These findings offer knowledge of the role that each of these receptors plays in the inflammatory response in the bovine and provide insight into the potential mechanisms that may be affected in neutrophils of cows with different CXCR1 genotypes. PMID- 17119954 TI - Bovine and water buffalo Mx2 genes: polymorphism and antiviral activity. AB - Millennia-long selective pressure of single-strand RNA viruses on the bovine Mx locus has increased the advantages of using the bovine Mx protein to evaluate the ultimate significance of the antiviral role of Mx proteins. The conclusions of research based only on the bovine Mx1 protein showed the need for comprehensive studies that demonstrate the role of all isoforms, individually or together, especially in the presence of a second isoform, the bovine Mx2 gene. This study provides information about bovine and water buffalo Mx2 genes, as well as their allelic polymorphism and basic antiviral potential. Observation of an Mx2 cDNA sequence (2,381 bp) obtained from 15 animals from 11 breeds using primers based on a previous sequence (NCBI accession no. AF335147) revealed several nucleotide substitutions, with eight different alleles and two amino acid exchanges: Gly to Ser at position 302 and Ile to Val at position 354, though the latter was found only in the NCBI database. A water buffalo Mx2 cDNA sequence was identified for the first time, revealing 46 nucleotide substitutions with 12 amino acid variations, in addition to a 9-bp insertion in the 5' untranslated region UTR, compared with the bovine Mx2 cDNA. Transfected 3T3 cells expressing bovine Mx2 mRNAs coding Gly or Ser at position 302, water buffalo Mx2 mRNA, positive control bovine Mx1 mRNA-expressing cells, and negative control parental 3T3 were subjected to infection with recombinant vesicular stomatitis virus (VSVDeltaG* G), as were empty pCI-neo vector-transfected cells. The positive control and all cells expressing Mx2 mRNAs displayed significantly higher levels of antiviral activity against VSVDeltaG*-G (P < 0.01) than did the negative controls. PMID- 17119955 TI - Screening and its potential application of lipolytic activity from a marine environment: characterization of a novel esterase from Yarrowia lipolytica CL180. AB - To develop an enantioselective lipase/esterase hydrolyzing racemic ofloxacin ester to levofloxacin, samples were collected from a variety of marine environments such as cold sea, hydrothermal vent area, sediment, tidal flat area, arctic sea, marine organisms, and so on. Microorganisms were isolated by plating on an enrichment medium with simultaneous detection of lipolytic activities and screened for the hydrolysis of ofloxacin ester. Three candidates among isolates were selected, and one of them, identified as Yarrowia lipolytica CL180, hydrolyzed preferentially S-enantiomer of racemic ofloxacin ester. The lipase/esterase gene (yli180) was cloned by screening a genomic library. The sequence analysis revealed an open reading frame consisting of 1,431 bp that encoded a protein of 476 amino acids with a molecular mass of 53 kDa. The yli180 gene was expressed in Escherichia coli and purified to homogeneity. The optimum activity of the recombinant protein (rYli180) occurred at pH 7.5 and 35 degrees C, respectively. rYli180 preferentially hydrolyzed p-nitrophenyl esters of fatty acids with short chain lengths of < or =10 carbon atoms. This study represents a novel esterase of type B1 carboxylesterase/lipase family from a marine isolate, showing a potential usage as a biocatalyst because of enantioselectivity toward racemic ofloxacin ester. PMID- 17119956 TI - Overview of some theoretical approaches for derivation of the Monod equation. AB - The Monod equation has been widely applied to describe microbial growth, but it has no mechanistic basis and is purely empirical. Extensive efforts have been dedicated to develop theoretical approaches for derivation of the Monod equation, which can be classified into three major groups, i.e., kinetic, thermodynamic, and substance transport approaches. In this review, four representative approaches are thus discussed. Due to the fact that different assumptions are made in each approach, no universal physical meaning of the Monod constant (K (s)) can be revealed. However, it seems that the Monod constant would be free energy-dependent and have nonequilibrium thermodynamic characteristics. PMID- 17119957 TI - Medium optimization by combination of response surface methodology and desirability function: an application in glutamine production. AB - An optimization strategy based on desirability function approach (DFA) together with response surface methodology (RSM) has been used to optimize production medium in L-glutamine fermentation. Fermentation problems often force to reach a compromise between different experimental variables in order to achieve the most suitable strategy applying in industrial production. The importance of the use of multi-objective optimization methods lies in the ability to cope with this kind of problems. A sequential RSM with different combinations of glucose and (NH(4))(2)SO(4) was performed to attain the optimal medium (OM-1) in glutamine production. Based on the result of RSM and the evaluation of production cost, a more economical optimal medium (OM-2) was obtained with the aid of DFA. In DFA study, glutamate, the main by-product in glutamine fermentation as another response was considered. Compared with OM-1 in validated experiment, similar amounts of glutamine were obtained in OM-2 while the concentration of glutamate and the production cost decreased by 53.6 and 7.1%, respectively. PMID- 17119958 TI - Multiple antibiotic resistances of Enterococcus isolates from raw or sand filtered sewage. AB - Fifty antibiotic-resistant Enterococcus strains were isolated from raw sewage of a wastewater treatment plant and from the same sewage after trickling through a 25-cm sand column, which retained >99% of the initial population. All 50 Enterococcus isolates were resistant against triple sulfa and trimethoprim/sulfamethoxazole and none were resistant against vancomycin. Most of the isolates from raw sewage were resistant to more antibiotics than the isolates from sand column effluent. One Enterococcus isolate from raw sewage (no. 61) and one Enterococcus isolate from sand column effluent (no. 95) had ten antibiotic resistances each. Isolate no. 95 maintained its resistances in the absence of antibiotics during the whole study. It was compared with isolate no. 70, which was one of the isolates, being resistant only against the two sulfonamides. Phenotypically and biochemically, the two organisms were strains of Enterococcus faecalis. Sequence analysis of partical 16S rDNA allowed alignment of isolate no. 95 as a strain of Enterococcus faecium and of isolate no. 70 as a strain of E. faecalis. E. faecium strain no. 95 carried at least six different plasmids, whereas for E. faecalis strain no. 70, no discrete plasmid band was seen on the gels. PMID- 17119959 TI - Performance evaluation of the Philips MOSAIC small animal PET scanner. AB - PURPOSE: In this study an evaluation of the performance of the Philips MOSAIC small animal PET scanner is presented, with special emphasis on the ability of the system to provide quantitatively accurate PET images. METHODS: The performance evaluation was structured according to NEMA-like procedures. RESULTS: The transaxial spatial resolution of the system (radial component) ranged between 2.7 mm FWHM at the centre and 3.2 mm FWHM at a radial offset of 45 mm from the centre. The axial spatial resolution of the system ranged between 3.4 mm FWHM at the centre and 5.8 mm FWHM at a radial offset of 45 mm from the centre. The scatter fraction was determined for a mouse- as well as for a rat-sized phantom, and the values obtained were 9.6% and 16.8%, respectively. For the mouse phantom, the maximum count rate measured was 560 kcps at 93 MBq; the maximum NEC rate equalled 308 kcps at 1.7 MBq/ml. For the rat phantom, these values were 400 kcps at 100 MBq and 129 kcps at 0.24 MBq/ml, respectively. The sensitivity of the system was derived to be 0.65%. An energy window between 410 and 665 keV was used in all experiments. CONCLUSION: The MOSAIC system exhibits moderate spatial resolution and sensitivity values, but good NEC performance. In combination with its relatively large field of view, the system allows for high-throughput whole body imaging of mice and rats. The accurate measurement of relative changes in radiotracer distributions is feasible. PMID- 17119960 TI - Juvenile fibroadenoma of the breast demonstrated on 111in-octreotide SPECT and 18F-FDG PET/CT. PMID- 17119961 TI - Early mechanical failure in total knee arthroplasty. AB - Early mechanical dysfunction of a total knee arthroplasty (TKA) is a challenging problem in terms of causality and solutions. The current strategy in our department is to perform a complete TKA revision rather than the less invasive partial procedures when a clear mechanical cause of failure has been found. In this investigation, we assessed 21 patients who underwent complete TKA revision in 2003-2004 in our institution within the first two years following the index TKA. Various clinical presentations included pain, stiffness, instability, and femoro-patellar signs. These corresponded to implant size, position, and fixation issues. The IKS knee score/function significantly increased from 47/47 to 85/78 at follow-up (six months minimum). Compared to the data in the literature, this systematic full revision seems to be a reasonable approach. This attitude takes advantage of the modularity of the implants for allowing perioperative adjustments of position, fixation, and constraint. Based on the results of our study, we propose a list of six mechanical pitfalls to be evaluated in the case of early dysfunction: frontal misalignment, sagittal overstuffing or malpositioning, axial malrotation, poor bone fixation, inappropriate constraint or ligamentous balance, and inappropriate level of the joint space. PMID- 17119962 TI - Surgical versus non-surgical treatment of chronic low back pain: a meta-analysis of randomised trials. AB - We performed a meta-analysis of randomised controlled trials to investigate the effectiveness of surgical fusion for the treatment of chronic low back pain compared to non-surgical intervention. Several electronic databases (MEDLINE, EMBASE, CINAHL and Science Citation Index) were searched from 1966 to 2005. The meta-analysis comparison was based on the mean difference in Oswestry Disability Index (ODI) change from baseline to the specified follow-up of patients undergoing surgical versus non-surgical treatment. Of the 58 articles identified, three studies were eligible for primary analysis and one study for sensitivity analysis, with a total of 634 patients. The pooled mean difference in ODI between the surgical and non-surgical groups was in favour of surgery (mean difference of ODI: 4.13, 95%CI: -0.82 to 9.08, p = 0.10, I(2) = 44.4%). Surgical treatment was associated with a 16% pooled rate of early complication (95%CI: 12-20, I(2) = 0%). Surgical fusion for chronic low back pain favoured a marginal improvement in the ODI compared to non-surgical intervention. This difference in ODI was not statistically significant and is of minimal clinical importance. Surgery was found to be associated with a significant risk of complications. Therefore, the cumulative evidence at the present time does not support routine surgical fusion for the treatment of chronic low back pain. PMID- 17119963 TI - Limited effect of rituximab on thrombocytopaenia and anticardiolipin antibodies in a patient with primary antiphospholipid syndrome. PMID- 17119964 TI - Whole blood laboratory model of thrombocytopenia for use in evaluation of hemostatic interventions. AB - This study describes a laboratory model of whole blood (WB) thrombocytopenia established with blood from healthy volunteers. We obtained a mean platelet count of 16 x 10(9)/l (95% confidence interval, 10-22) in WB by repeatedly replacing the platelet-rich supernatant with autologous platelet-poor plasma from the same individual. Thrombelastographic profiles of WB clot formation and WB clot stability were performed in parallel with measurements of WB platelet aggregation response. Thrombocytopenia reduced the maximum rate of WB clot formation, while ex vivo addition of platelets reversed the coagulopathy of thrombocytopenia. Control experiments revealed minimal changes in coagulation factors, distribution of bloods cells, and platelet activation capabilities. The WB model appears useful in research, development, and evaluation of the effects of hemostatic interventions in thrombocytopenia. PMID- 17119965 TI - Usefulness of the MSG/IFICG/EORTC diagnostic criteria of invasive pulmonary aspergillosis in the clinical management of patients with acute leukaemia developing pulmonary infiltrates. AB - Invasive pulmonary aspergillosis (IPA) is a frequently fatal complication in patients with acute leukaemia. Because diagnosis is still difficult, non-invasive diagnostic criteria were recently proposed by MSG/IFICG/EORTC for study purposes. We have analysed their usefulness in the clinical management of acute leukaemic patients with pulmonary infiltrates. Twenty-seven infiltrates developed during 174 chemotherapy cycles given to 50 consecutive patients. According to diagnostic criteria, IPA was diagnosed in 42% of patients and 77.8% of pulmonary infiltrates. AML diagnosis and the first induction cycle were significant risk factors. "Proven" IPA was rare, occurring in one patient (2%). The diagnosis of "probable" IPA was made in seven patients (14%) and was strongly supported by the significant association of characteristic radiological lesions ("major" clinical criterion) with the positivity of one microbiological criterion (P = 0.026). Conversely, "possible" IPA was frequent (26%) because its pertinent diagnostic criteria were fulfilled in 48.1% of pulmonary infiltrates. However, in 84.6% of cases, the diagnosis of "possible IPA" aspecifically derived from the association of two conditions, a new pulmonary infiltrate with symptoms of lower respiratory tract infection ("minor clinical criterion"), together with the definition of "susceptible" host, which applied to 100% of our leukaemic patients. We conclude that, according to MSG/IFICG/EORTC criteria, a high number of pulmonary infiltrates would be diagnosed as IPA, but only a diagnosis of "proven/probable" IPA should be considered reliable in the clinical management of suspected IPA. PMID- 17119966 TI - Mechanisms of acquired chemoresistance to 5-fluorouracil and tomudex: thymidylate synthase dependent and independent networks. AB - PURPOSE: Thymidylate synthase (TS) over-expression is widely accepted as a major molecular mechanism responsible for 5-fluorouracil (5-FU) and tomudex (TDX) resistance. In this study, the importance of TS in 5-FU and TDX resistance was evaluated. METHODS: The sensitivity of TS-over-expressing 5-FU (3) and TDX (3) resistant cell lines to 5-FU and TDX was analysed. The cross-resistance between 5 FU and TDX resistant cell lines was determined. The relationship between p53 and NF-kappaB status and the sensitivity to 5-FU and TDX was evaluated. RESULTS: Compared to relevant parental sensitive cell lines, the 5-FU resistant cell lines were highly cross-resistant to TDX (over 20,000-fold). In contrast, over expression of TS did not significantly confer 5-FU resistance on the TDX resistant cell lines (0.8- to 1.3-fold). Thymidine (20 microM) rescue induced TDX resistance in TDX sensitive cell lines (over 10,000-fold) but only moderately influenced 5-FU sensitivity in 5-FU sensitive cell lines (1.1- to 2.4-fold). Uridine moderately protected one cancer cell line (RKO) from 5-FU-induced, but not TDX-induced, cytotoxicity. NF-kappaB transfected MCF-7 and p53 knockout HCT116 cells were resistant to 5-FU (4.4- and 2.4-fold, respectively) but not to TDX. TS protein expression in NF-kappaB transfected and p53 knockout cell lines was comparable to the relevant parental cell lines. CONCLUSION: In some cancer cell lines, TS-independent molecular events may play a key role in 5-FU resistance. Loss of p53 function and NF-kappaB over-expression may be involved in TS-independent 5-FU chemoresistance in some cancer cell lines. PMID- 17119967 TI - Analysis of a certain class of replicator equations. AB - Motivated by a problem in the evolution of sensory systems where gains obtained by improvements in detection are offset by increased costs, we prove several results about the dynamics of replicator equations with an n x n game matrix of the form: A( ij ) = a( i )b( j ) - c( i ). First, we show that, generically, for this class of game matrix, all equilibria must be on the 1-skeleton of the simplex, and that all interior solutions must limit to the boundary. Second, for the particular ordering, a1b2> ... >bn, which is most natural in the study of the evolution of sensory systems, we show that topological restrictions require a unique local attractor in every face of the simplex. We conjecture that the unique local attractor for the full simplex is, in fact, a global attractor, and prove this for n < or = 5. In a separate argument supporting the conjecture, we show that there can be no chain recurrent invariant set entirely contained in the 1-skeleton of the simplex. Finally, we discuss the special, non-generic case and give a local description of the dynamics when there is an interior equilibrium. PMID- 17119968 TI - A chitinase gene, chiB, involved in the autolytic process of Aspergillus nidulans. AB - Chitinases are thought to be involved in the morphogenesis and autolysis of filamentous fungi. We cloned a gene (chiB) encoding a class V chitinase from Aspergillus nidulans. ChiB expressed in Escherichia coli had chitin-hydrolyzing activity, indicating that chiB encoded a chitinase. Deletion of chiB affected neither germination efficiency nor hyphal growth rate, but considerably reduced the intracellular and extracellular chitinase activities. The decrease in hyphal dry weight during autolytic phase was slower in the mutant than in the wild-type strain. Western blot analysis indicated that the quantity of ChiB significantly increased when the wild-type mycelia were starved for carbon sources, a condition that induced hyphal autolysis. These results suggest that chiB plays an important role in the autolytic process in A. nidulans. PMID- 17119969 TI - Proteome mapping of the Trichoderma reesei 20S proteasome. AB - The 26S proteasome, a multicatalytic protease comprising the catalytic 20S core particle and the 19S regulatory particle has a crucial role in cellular protein quality control. We have used a chromatography-based approach to purify and map the protein content of the 20S core particle from the industrially-exploited filamentous fungus Trichoderma reesei. There are no previous reports on the isolation or proteomic mapping of the proteasome from any filamentous fungus. From the reference map, 13 of the 14 20S proteasome subunits and many related proteins that co-purified with the 20S proteasome have been identified. These include 78 kDa glucose-regulated protein (BIP) and several chaperones including heat shock proteins involved in the unfolded protein response (UPR). Some proteasome interacting proteins (PIPs) were also identified on the proteome map and included 14-3-3-like protein, glyceraldehyde-3-phosphate dehydrogenase, transaldolase, actin, translation elongation factor, enolase, ATPase in the ER (CDC48), and eukaryotic initiation factor. We present here a master map for the 20S catalytic core to pave the way for future differential display studies addressing intracellular degradation of endogenous and foreign proteins in filamentous fungi. PMID- 17119970 TI - Effects of pravastatin in murine collagen-induced arthritis. AB - Here we evaluated whether 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) have beneficial effects for collagen-induced arthritis (CIA). DBA/1 mice were immunized with bovine type-II collagen and administered 100 mg/kg of pravastatin interperitoneally. We measured the effects of pravastatin for CIA including infiltration of macrophages at the synovial membrane and production of anti-type-II collagen antibodies and cytokines. Adverse reactions of pravastatin were also measured. The pravastatin-treated mice had delayed onset of CIA compared with the controls. The involvement of inflammatory cells in the synovial membrane and the expression of monocyte chemotactic protein-1 (MCP-1) mRNA in the joint were reduced. Moreover, some cytokines (TNF-alpha, IL-6, IFN-gamma) and MCP-1 levels in the supernatants of spleen cells cultured with pravastatin decreased. Meanwhile, adverse reactions of pravastatin, such as peritonitis, were not detected. Pravastatin may have good prospects for treating some anti-inflammatory effects on human rheumatoid arthritis. PMID- 17119971 TI - Quantitative and qualitative assessment of hand function and deformity in systemic sclerosis. PMID- 17119972 TI - Irreversible JPEG 2000 compression of abdominal CT for primary interpretation: assessment of visually lossless threshold. AB - To estimate the visually lossless threshold for Joint Photographic Experts Group (JPEG) 2000 compression of contrast-enhanced abdominal computed tomography (CT) images, 100 images were compressed to four different levels: a reversible (as negative control) and irreversible 5:1, 10:1, and 15:1. By alternately displaying the original and the compressed image on the same monitor, six radiologists independently determined if the compressed image was distinguishable from the original image. For each reader, we compared the proportion of the compressed images being rated distinguishable from the original images between the reversible compression and each of the three irreversible compressions using the exact test for paired proportions. For each reader, the proportion was not significantly different between the reversible (0-1%, 0/100 to 1/100) and irreversible 5:1 compression (0-3%). However, the proportion significantly increased with the irreversible 10:1 (95-99%) and 15:1 compressions (100%) versus reversible compression in all readers (P < 0.001); 100 and 95% of the 5:1 compressed images were rated indistinguishable from the original images by at least five of the six readers and all readers, respectively. Irreversibly 5:1 compressed abdominal CT images are visually lossless and, therefore, potentially acceptable for primary interpretation. PMID- 17119973 TI - Cine MR imaging of uterine peristalsis in patients with endometriosis. AB - Endometriosis is one of the most important causes of infertility; however the precise mechanism by which it affects female fertility is unclear. The objective of this study was to study the functional aspects of the uterus by evaluating uterine contractility in patients with endometrial cysts of the ovary. The study population was recruited from two institutes and consisted of 26 women (periovulatory (10), luteal (13), and menstrual phase (3); age range: 19-51 years) with untreated endometriosis; the control group consisted of 12 healthy women (age range: 22-41 years). Cine MR imaging obtained by a 1.5T magnet was visually evaluated at 12x faster than real speed, focusing on the presence of uterine peristalsis, the direction and frequency of peristalsis, and the presence of sustained uterine contractions. Uterine peristalsis was identifiable in 3/10, 3/13, and 3/3 of the endometriosis patients in each menstrual cycle, respectively, and in 11/12, 3/12, and 5/12 of their control subjects. Peristaltic detection rate and frequency were significantly less for the endometriosis group than for the controls in the periovulatory phase only (p<0.05). Sustained contractions were recognized in 19/36 control subjects and in 13/26 endometriosis patients, but the difference was not significant. Uterine peristalsis appears to be suppressed during the periovulatory phase in patients with endometriosis, which may have an adverse effect on sperm transport. PMID- 17119974 TI - Color Doppler, power Doppler and B-flow ultrasound in the assessment of ICA stenosis: Comparison with 64-MD-CT angiography. AB - The purpose of this study is to investigate the diagnostic potential of color coded Doppler sonography (CCDS), power-Doppler (PD) and B-flow ultrasound in assessing the degree of extracranial internal carotid artery (ICA) stenosis in comparison to CT-angiography (MD-CTA). Thirty-two consecutive patients referred for CTA with 41 ICA-stenoses were included in this prospective study. MD-CTA was performed using a 64 row scanner with a CTDIvol of 13.1 mGy/cm. In CTA, CCDS, PD and B-flow, the degree of stenosis was evaluated by the minimal intrastenotic diameter in comparison to the poststenotic diameter. Two radiologists performed a quantitative evaluation of the stenoses in consensus blinded to the results of ultrasound. These were correlated to CTA, CCDS, PD and B-flow, intraoperative findings and clinical follow-up. Grading of the stenoses in B-flow ultrasound outperformed the other techniques in terms of accuracy with a correlation coefficient to CTA of 0.88, while PD and CCDS measurements yield coefficients of 0.74 and 0.70. Bland-Altman analysis additionally shows a very little bias of the three US methods between 0.5 and 3.2 %. There is excellent correlation (coefficient 0.88, CI 0.77-0.93) with 64-MD-CTA and B-flow ultrasound in terms of accuracy for intrastenotic and poststenotic diameter. Duplex sonography is useful for screening purposes. PMID- 17119976 TI - Larval lampreys possess a functional lateral line system. AB - Morphology of larval lampreys' neuromasts was found to be very similar to that of adults. Activity in the lateral line nerve, elicited by a vibrating ball, indicated a functional lateralis system. Analysis revealed at least two populations of afferents, responding to opposite directions of water flow, with adapting responses. The response magnitude increased monotonically with stimulus amplitude. Larval lampreys' neuromasts were less sensitive than those of teleosts. At low frequencies the response showed a phase lead of 200-220 degrees with respect to the maximum of the ball displacement and a gain that was approximately linearly proportional to frequency. PMID- 17119975 TI - A comparison of primary two- and three-dimensional methods to review CT colonography. AB - The aim of our study was to compare primary three-dimensional (3D) and primary two-dimensional (2D) review methods for CT colonography with regard to polyp detection and perceptive errors. CT colonography studies of 77 patients were read twice by three reviewers, first with a primary 3D method and then with a primary 2D method. Mean numbers of true and false positives, patient sensitivity and specificity and perceptive errors were calculated with colonoscopy as a reference standard. A perceptive error was made if a polyp was not detected by all reviewers. Mean sensitivity for large (> or = 10 mm) polyps for primary 3D and 2D review was 81% (14.7/18) and 70%(12.7/18), respectively (p-values > or = 0.25). Mean numbers of large false positives for primary 3D and 2D were 8.3 and 5.3, respectively. With primary 3D and 2D review 1 and 6 perceptive errors, respectively, were made in 18 large polyps (p = 0.06). For medium-sized (6-9 mm) polyps these values were for primary 3D and 2D, respectively: mean sensitivity: 67%(11.3/17) and 61%(10.3/17; p-values > or = 0.45), number of false positives: 33.3 and 15.6, and perceptive errors : 4 and 6 (p = 0.53). No significant differences were found in the detection of large and medium-sized polyps between primary 3D and 2D review. PMID- 17119977 TI - Delayed rupture of traumatic aneurysm after civilian craniocerebral gunshot injury in children. AB - BACKGROUND: There are few published large series on civilian craniocerebral gunshot injuries in children. Traumatic intracranial aneurysms (TICAs) are rare and highly unstable lesions. They represent less than 1% of all aneurysms and can either rupture within minutes after formation or remain quiescent for several weeks or years, manifesting with delayed hemorrhage and neurologic deterioration. CASE HISTORY: We report the case of a 10-year-old girl who was referred for coma after high-velocity craniocerebral gunshot wound and neurological deterioration 7 days after the initial injury. A massive right posterior occipital hematoma caused by the rupture of an unsuspected right posterior cerebral artery TICA was discovered. TICA was treated by coil embolization, with a good neurological recovery at 6-month follow-up. DISCUSSION: We discuss the pathogenesis and the management of TICA in a child after civilian craniocerebral gunshot injuries. CONCLUSIONS: TICAs should be suspected in patients with civilian craniocerebral gunshot injuries, presenting with secondary neurological deterioration, to carry out emergent CT scan and angiographic exploration before contemplating definitive endovascular treatment. Endovascular management may be a prompt safe-to-use technique and a valuable option, especially when surgery is highly risky. PMID- 17119978 TI - Outcome of children with posterior fossa medulloblastoma: a single institution experience over the decade 1994-2003. AB - AIM: While the impact of radiotherapy in the management of medulloblastoma was recognised, the introduction of chemotherapy was investigated in clinical trials and shown to confer an additional advantage. We reviewed the outcome of a series of consecutive patients to assess the impact in a population-based clinical establishment. MATERIALS AND METHODS: A series of 38 children treated for medulloblastoma at Birmingham Children's Hospital between 1994 and 2003 was analysed. The effect of surgery, radiotherapy, chemotherapy and metastasis on survival was analysed. RESULTS: The overall 5-year survival rate was 61.4% for the 36 patients who had resective surgery, while 2 patients had biopsy only and died within a few months. There was no operative mortality. The incidence of hydrocephalus needing permanent shunting was higher in the first 3 years of life (p = 0.007, chi-square). The 5-year survival rate of patients with total and sub total excision of medulloblastoma was 61.1% and 61.8%, respectively. The 5-year survival rate of patients older than 3 years was 73.4% and for patients under 3 years was 36.3% (p = 0.007, log rank). Metastases at presentation did not influence survival. All deaths occurred in the first 32 months. CONCLUSION: The contribution of chemotherapy in the improvement of the overall survival appears more evident in children younger than 3 years or presenting with metastases. The absence of significant difference in survival between patients with total or sub total excision of medulloblastoma supports the view that total excision of medulloblastoma can be avoided when the risk for potential intra-operative damage and consequent neurological deficits is high. PMID- 17119979 TI - Pediatric intracranial aneurysms-clinical characteristics and outcome of surgical treatment. AB - OBJECT: Intracranial aneurysms are rare in children and have different epidemiological, clinical, and morphological characteristics and outcome from those in adults. MATERIALS AND METHODS: We analyzed demographic, clinical, radiologic features, treatment, and outcome in 55 patients <18 years of age, treated from Jan 1995 through December 2005. RESULTS: Intracranial aneurysms in children below 18 years constituted 4% of all intracranial aneurysms. Internal Carotid artery (ICA) bifurcation was the commonest location. About half of the aneurysms were complex. Three-fourth of the patients required surgical treatment. Two patients died, constituting 5% mortality. Two patients (5%) had poor outcome, whereas the majority (90%) had a favorable outcome. CONCLUSIONS: Pediatric aneurysms have male predominance, higher incidence of clinical features of mass effect or seizures, high incidence of large, traumatic/mycotic aneurysms, associated illnesses and ICH/IVH and hydrocephalus, better Hunt and Hess grades at presentation, ICA bifurcation as the commonest site, and better outcome than their adult counterparts. PMID- 17119980 TI - Histological findings in cerebellar tonsils of patients with Chiari type I malformation. AB - OBJECTIVES: Cerebellar tonsillectomy is often performed for relief of symptoms associated with Chiari type I malformation (CMI). Nonetheless, the idea of removing supposedly healthy central nervous tissue has been a source of concern for neurosurgeons. The aim of this paper is to determine the histological changes in the cerebellar tonsils of patients with a wide range of symptoms and conditions related to CMI. MATERIALS AND METHODS: The cerebellar tonsils of 43 pediatric patients with CMI were sent to pathology for histological examination. CONCLUSION: The cerebellar tonsils in a great majority of CMI patients can be abnormal. We suggest that the reported histological findings are secondary to injury and ischemia. PMID- 17119981 TI - Treatment outcomes for hepatoblastoma: an institution's experience over two decades. AB - Our objective is to review a leading Australian tertiary referral centre's experience in treating hepatoblastoma over two decades. This is a retrospective study of clinical, radiological and pathological data in hepatoblastoma patients treated at our institution between 1984 and 2004. Thirty children (17 male, 13 female) ranging in age from 5 months to 6.5 years (median 19 months) at diagnosis were reviewed. This is the largest case series reported in Australia to date. The median length of follow-up was 6.3 years (range 7 months to 20.9 years). On average, the number of new cases increased by 0.12 each year (P=0.01). A total of 29 patients underwent definitive surgery for primary tumour control. Of these, two (7%) had initial primary surgery, while 27 (93%) had delayed surgery following neo-adjuvant chemotherapy. Chemotherapy regimens included the SIOPEL study protocols. Patients not enrolled in SIOPEL tended to be given more courses of chemotherapy. Ten patients (34%) received an extended right hemi-hepatectomy, six (21%) had right hemi-hepatectomy, seven (24%) had left hemi-hepatectomy, three (10%) had left lateral segmentectomy, and two (7%) had a non-anatomical resection. In addition, two patients required IVC reconstruction at the time of their primary liver resection. Overall survival at 5 years was 89.1% (95% CI 69.8 96.4%). Event-free survival at 5 years was 75.7% (95% CI 53.2-88.5%). There was a clear increase in the risk of recurrence (RR=4.8) and death (RR=4.5) where margins were not microscopically clear. However, neither reached statistical significance in this small cohort. Surgery was still worthwhile even when margins were involved. There was no correlation between mean AFP level at diagnosis, and outcome. Our experience suggests that the achievement of microscopically clear margins at the time of primary tumour resection is an important factor in achieving a positive long-term outcome. In addition, extended courses of neo adjuvant chemotherapy in patients with pulmonary metastases achieves good long term outcomes, provided adequate surgical resection of the primary tumour with clear microscopic margins can be achieved. PMID- 17119982 TI - Simple harmonic scalpel hemorrhoidectomy utilizing local anesthesia combined with intravenous sedation: a safe and rapid alternative to conventional hemorrhoidectomy. AB - BACKGROUND: Harmonic Scalpel(R) hemorrhoidectomy (HSH) is an established surgical therapy for the treatment of symptomatic grade III and IV hemorrhoids. Hemorrhoid surgery is still being performed as an inpatient procedure with general or regional anesthesia in many centers today. There was a trend toward performing hemorrhoid surgery as an ambulatory procedure using local anesthesia supplemented with intravenous sedation. The aim of the current study was to evaluate the safety and efficacy of HSH performed with combination local anesthesia and intravenous sedation in an ambulatory surgical center. MATERIALS AND METHODS: A retrospective review was performed on the clinical charts of all patients undergoing HSH in an ambulatory surgical center from 2001 to 2005. All hemorrhoidectomies were attempted under propofol/ketamine intravenous sedation and local anesthesia in the prone position. A simple, open technique without routine suture was used. RESULTS: During the study period, 180 patients (70 females) underwent HSM. Mean procedure and total operating room time were 12 and 28 min, respectively. One patient (0.6%) was converted to general endotracheal anesthesia. Ten patients (5.6%) required post anesthesia care unit (PACU) observation. All patients were discharged home after the procedure. Postoperative complications occurred in 19 patients (10.6%). There were no reoperations and the total readmission rate was 3.7%. CONCLUSION: HSH performed with a combination of intravenous sedation and local anesthesia is safe and effective in the ambulatory surgery setting. The combined technique was associated with a rate of complications comparable to published series utilizing conventional hemorrhoidectomy techniques. Added benefits include shorter hospital stay and a potential for cost savings. PMID- 17119983 TI - Peritoneal instillation of taurolidine or polihexanide modulates intestinal microcirculation in experimental endotoxemia. AB - BACKGROUND AND AIMS: Treatment of peritonitis may include peritoneal lavage/instillation with anti-infective agents like taurolidine or chlorhexidine. MATERIALS AND METHODS: We examined the effects of peritoneal instillation (INST, 5-ml solution) with taurolidine (TAURO) or polihexanide (POLI-LS) on intestinal microcirculation using intravital microscopy (IVM) in experimental endotoxemia (15 mg/kg lipopolysaccharide i.v.; LPS) in the rat (n = 8 each group), their direct effects on local small blood vessels, aortal rings, and myocardial strips in vitro, as well as plasma interleukin levels. RESULTS: It was found that LPS produced hypotension (98.8 +/- 9.5 vs 130.4 +/- 10.5 mmHg; mean arterial pressure [MAP], mean +/- standard deviation [SD]), which was further pronounced after INST of TAURO (78.8 +/- 10.8; P < 0.005) or POLI-LS (78.1 +/- 6.0; P < 0.001). IVM revealed a reduction in temporary adhering leucocytes and an increase in firmly adhering leucocytes after INST with TAURO and POLI-LS. Both agents reduced functional capillary density either in the mucosa (POLI-LS vs sham: 259.7 +/- 54 cm/cm(2) vs 337.1 +/- 35.5) or longitudinal muscular layer in LPS rats (TAURO vs sham: 119.8 +/- 14.8 vs 153.7 +/- 11.0). POLI-LS induced local vasodilatation, whereas TAURO induced small vasoconstriction; in vitro, both agents showed vasodilating properties and did not have any effect on myocardial strip contraction. CONCLUSION: Some of the observed microcirculatory changes could be a result of the direct vascular effects of these agents. PMID- 17119984 TI - Transureteroureterostomy allows renal sparing radical resection of advanced malignancies with rectosigmoid invasion. AB - BACKGROUND: In case of advanced colorectal cancer or other malignancies with pelvic involvement, tumor invasion of the ureter may afford partial ureteral resection for curative multivisceral resection. One preferable approach for the primary reconstruction of the urinary tract to preserve the ipsilateral kidney and maintain normal urinary function is transureteroureterostomy, i.e. termino lateral anastomosis of both ureters. MATERIALS AND METHODS: Between June 2001 and August 2004, 15 multivisceral en bloc resections including a distal tumor infiltrated ureteral segment were performed. Reconstruction was achieved by transureteroureterostomy with or without additional insertion of double J stents. Clinical outcome and especially complications of the urinary tract were monitored within the follow-up-period until August 2005. RESULTS: Post-operative course was uncomplicated in 12 cases and double J stents were removed after ultrasound control on the 14th post-operative day. Percutaneous nephrostomy was post operatively necessary in three patients who were initially operated without insertion of double J catheters: two suffered from leakage of the transureteroureterostomy and one patient had bilateral hydronephrosis due to stenosis of the transureteroureterostomy. After 6 weeks, percutaneous nephrostomy could be removed and urine flow was uncomplicated in all three patients. CONCLUSION: For patients with advanced colorectal cancer or other pelvic malignancy, transureteroureterostomy is a favourable technique for definitive reconstruction of the urinary tract whenever tumor resection affords partial ureteral resection. Intra-operative insertion of double J stents into both ureters is highly recommended to prevent leakage or stenosis of the anastomosis. PMID- 17119985 TI - Preoperative serum levels of c-erbB-2 do not seem to be useful in management of patients with rectal cancer. AB - BACKGROUND AND AIM: Soluble c-erbB-2 oncoprotein has been proven as a useful marker in the management of breast cancer patients, but its value in diagnostics and follow-up of colorectal cancer patients remains controversial. The aim of this study was to evaluate the usefulness of serum c-erbB-2 monitoring in diagnostics and prediction of disease outcome in rectal cancer patients. MATERIALS AND METHODS: Serum samples from 88 patients with rectal adenocarcinoma before surgery and from 41 healthy controls were tested for the presence of c erbB-2 oncoprotein by ELISA, and the patients were followed up for at least 5 years after the surgery. RESULTS: Preoperative serum c-erbB-2 levels were significantly higher in stage IV patients than in healthy controls (P<0.001) and did not show correlation with preoperative CEA levels. Elevated preoperative serum c-erbB-2 levels showed relatively high specificity (88%) and low sensitivity (44%) in the diagnosis of rectal cancer. Elevated preoperative oncoprotein levels were predictive neither for overall survival nor for development of local recurrence/distant metastases. CONCLUSION: Although preoperative serum c-erbB-2 levels were significantly higher in rectal cancer patients than in healthy controls, the soluble c-erbB-2 does not seem to be useful in the diagnosis of rectal cancer due to its low sensitivity. Preoperative serum levels of this oncoprotein were predictive neither for overall survival nor for local recurrence/distant metastases in rectal cancer patients. PMID- 17119986 TI - Low grade malignant peripheral nerve sheath tumor with smooth muscle differentiation. AB - Malignant peripheral nerve sheath tumors are usually Schwann cell derived. Differentiation along mesenchymal lines is uncommon. Herein we present an example with smooth muscle differentiation occurring in the left distal forearm of a 62 year-old male with neurofibromatosis type I. Incisional biopsy of the slowly growing mass demonstrated a low-grade malignant peripheral nerve sheath tumor with strong S-100 protein immunoexpression as well as focal smooth muscle actin staining in atypical neoplastic cells. Immunostains for epithelial membrane antigen, neurofilament protein, glial fibrillary acidic protein, CD57, desmin, and myogenin were negative. The MIB-1 labeling index was 5.4%. Electron microscopy revealed arrays of microfilaments (actin myofilaments) variably associated with fusiform dense bodies, plasmalemmal micropinocytotic vesicles, and foci of external lamina (basement membrane) consistent with smooth muscle differentiation. Smooth muscle should be added to the spectrum of differentiation exhibited by malignant peripheral nerve sheath tumors. Its recognition requires exclusion of the alternative diagnosis of leiomyosarcoma. PMID- 17119987 TI - Prediction of preservative sensitization potential using surface marker CD86 and/or CD54 expression on human cell line, THP-1. AB - Preservatives are important components in many products, but have a history of purported allergy. Several assays [e.g., guinea pig maximization test (GPMT), local lymph node assay (LLNA)] are used to evaluate allergy potential of preservatives. We recently developed the human Cell Line Activation Test (h CLAT), an in vitro skin sensitization test using human THP-1 cells. This test evaluates the augmentation of CD86 and CD54 expression, which are key events in the sensitization process, as an indicator of allergy following treatment with test chemical. Earlier, we found that a sub-toxic concentration was needed for the up-regulation of surface marker expression. In this study, we further evaluate the capability of h-CLAT to predict allergy potential using eight preservatives. Cytotoxicity was determined using propidium iodide with flow cytometry analysis and five doses that produce a 95, 85, 75, 65, and 50% cell viability were selected. If a material did not have any cytotoxicity at the highest technical dose (HTD), five doses are set using serial 1.3 dilutions of the HTD. The test materials used were six known allergic preservatives (e.g., methylchloroisothiazolinone/methylisothiazolinone, formaldehyde), and two non allergic preservatives (methylparaben and 4-hydroxybenzoic acid). All allergic preservatives augmented CD86 and/or CD54 expression, indicating h-CLAT correctly identified the allergens. No augmentation was observed with the non-allergic preservatives; also correctly identified by h-CLAT. In addition, we report two threshold concentrations that may be used to categorize skin sensitization potency like the LLNA estimated concentration that yield a three-fold stimulation (EC3) value. These corresponding values are the estimated concentration which gives a relative fluorescence intensity (RFI) = 150 for CD86 and an RFI = 200 for CD54. These data suggest that h-CLAT, using THP-1 cells, may be able to predict the allergy potential of preservatives and possibility classify the potency of an allergen. PMID- 17119988 TI - Twenty-four-hour CTG monitoring: comparison of normal pregnancies of 25-30 weeks of gestation versus 36-42 weeks of gestation. AB - OBJECTIVE: Routinely antepartal cardiotocogram (CTG) is recorded for 30 min in order to obtain normal resting phases (<30 min) or a decrease of irregulatory due to hypoxia (>30 min) or to differentiate these from each other. In case of early onset of hypoxia first pathological findings might only be seen by chance in incidentally recorded CTG. The goal of this study was, if a continuous 24-h CTG allows an earlier detection of beginning hypoxia in case of normal pregnancies of 36-42 weeks compared to pregnancies of 25-30 weeks of gestation, and if there are any differences between both groups concerning the qualitative and quantitative description or the detection of a circadian rhythm. METHODS: 21 patients in each group had 24-h CTGs by means of telemetry (Hewlett-Packard type 78101A, 80110A). In both study groups, fetal heart-rate tracing included a full qualitative and quantitative description. Comparison of the results of both groups was done to look for early signs of pathological findings concerning reduced fetal well-being and a potential day and night rhythm. RESULTS: In comparison to 36-42 weeks of pregnancies 25-30 weeks had significantly more physiological undulatory oscillation and less narrowed undulatory oscillation (P < 0.001), as well as less resting phases (P < 0.001). Baseline tachycardia and bradycardia showed significantly increasing quantity (P < 0.001). CONCLUSION: Twenty-four-hour CTG is a good screening method to detect early onset of hypoxia in case of second and third trimester pregnancies and a big help to detect a fetus at risk earlier. In future computerized CTG-systems should be proved in this connection and should be compared with our study. PMID- 17119989 TI - Creation of neovagina using oxidized cellulose (surgicel) as a surgical treatment of vaginal agenesis. AB - OBJECTIVE: Our goal was to analyze the safety and efficacy of the use of oxidized cellulose in the creation of neovagina. METHODS: A total of 10 women with vaginal agenesis attending gynaecology clinic of AIIMS between July 2003 to March 2005 underwent vaginal reconstructive surgery using oxidized cellulose were recruited in this retrospective study. The age of patients ranged from 17 to 22 years (mean age 19 years). Using a transverse incision at introitus, a 10 x 4 cm space was created and a vaginal mould covered with oxidized cellulose was put in the neovagina. RESULTS: The age ranged from 16 to 22 years (mean 19.5 years). Out of 10 patients, who had undergone vaginoplasty, all were sexually active; 8 had satisfactory intercourse, 1 chronic mild dyspareunia due to partial stenosis and 1 was lost to follow up. 2 out of 10 had inflammatory reaction and 1 had psychological problems. CONCLUSION: Vaginal reconstruction using oxidized cellulose appears to be a safe and effective procedure with minimum complications, morbidity and discomfort with high success rates. PMID- 17119990 TI - Preliminary multi-institutional prospective pathologic and molecular studies support preservation of sublevel IIB and level IV for laryngeal squamous carcinoma with clinically negative neck. AB - Level II-IV selective neck dissection, often performed bilaterally, has become the procedure of choice for elective dissection of the clinically negative (N0) neck in the treatment of laryngeal cancer. The most significant morbidity of this procedure is dysfunction of the accessory nerve, incurred by the necessity of mobilization and retraction of the nerve in order to remove the contents of sublevel IIB. Other morbidity includes possible injury to the phrenic nerve and chylous fistula. These complications are associated with the dissection of level IV. A number of prospective multi-institutional studies of the distribution of cervical lymph node metastases in the neck indicate that lymph nodes in sublevel IIB and level IV are rarely involved in cases of laryngeal cancer with N0 neck. Information was obtained by the study of neck dissection specimens by conventional light microscopy, and by molecular analysis of the specimens. Molecular analysis reveals a significant number of metastases that are not discovered by light microscopy, and is thus essential for this type of evaluation. The authors conclude that these preliminary studies indicate that it is safe and appropriate to eliminate dissection of sublevel IIB and level IV from the elective neck dissection performed for laryngeal cancer with N0 neck. This practice will reduce both operating time and morbidity, particularly accessory nerve dysfunction, without compromising the oncologic result. Further prospective studies are needed to confirm these conclusions. PMID- 17119991 TI - Sudden hearing impairment combined with diabetes mellitus or hyperlipidemia. AB - The objective of this study was to delineate whether the presence of the risk factors of microvascular disorder (MD) contribute to the prognostic outcome in patients with idiopathic sudden deafness (ISD). Retrospective clinical record review was conducted with 106 patients of ISD. Pure tone hearing thresholds were compared among the groups with or without hyperglycemia and hypercholesterolemia. Significantly better hearing recoveries were observed in cases with hyperglycemia as compared with controls. There was a tendency that the magnitude of hearing recovery in hypercholesterolemia was greater than that of controls although there was no statistical difference. The presence of the risks in MD could be an indicator of better hearing prognosis of ISD if we choose appropriate treatments. Heterogeneity of the ISD makes the establishment of the relationship between the risk factors of MD and etiology of some cases of ISD. We may have to choose the treatments for ISD considering whether the patient has any risk factor of MD. PMID- 17119992 TI - Preservation of large-scale chromatin structure in FISH experiments. AB - The nuclear organization of specific endogenous chromatin regions can be investigated only by fluorescence in situ hybridization (FISH). One of the two fixation procedures is typically applied: (1) buffered formaldehyde or (2) hypotonic shock with methanol acetic acid fixation followed by dropping of nuclei on glass slides and air drying. In this study, we compared the effects of these two procedures and some variations on nuclear morphology and on FISH signals. We analyzed mouse erythroleukemia and mouse embryonic stem cells because their clusters of subcentromeric heterochromatin provide an easy means to assess preservation of chromatin. Qualitative and quantitative analyses revealed that formaldehyde fixation provided good preservation of large-scale chromatin structures, while classical methanol acetic acid fixation after hypotonic treatment severely impaired nuclear shape and led to disruption of chromosome territories, heterochromatin structures, and large transgene arrays. Our data show that such preparations do not faithfully reflect in vivo nuclear architecture. PMID- 17119993 TI - Lack of association of the VEGF gene promoter (-634 G-->C and -460 C-->T) polymorphism and the risk of advanced retinopathy of prematurity. AB - BACKGROUND: Recently, it has been reported that genetic polymorphism (-634 G-->C and -460 C-->T) in the promoter region of the vascular endothelial growth factor (VEGF) gene can influence the progression of retinopathy of prematurity (ROP). In order to evaluate its general applicability as a screening procedure in clinics and to replicate the above result, we have undertaken the following study. METHODS: We have analyzed a cohort of 61 patients with advanced ROP (stage 4 and 5) along with 61 normal controls for the VEGF gene promoter polymorphism. For this purpose, blood samples were collected from each patient and leukocyte DNA was isolated. Genomic DNA was amplified by the polymerase chain reaction (PCR) method with two pairs of primers designed to amplify separately the promoter region (containing -634 G-->C and -460 C-->T polymorphism) of the VEGF gene. The amplified product was subjected to restriction enzyme digestion. The base change in the restriction site was further confirmed by a BigDye terminator cycle sequencing of the amplified product. RESULTS: Our analysis suggests that there is no significant difference in allelic frequency of the VEGF gene between normal subjects and patients with advanced ROP in our cohort. CONCLUSION: Our results do not support the association of the VEGF gene promoter polymorphism and the risk of advanced ROP. In order to adapt this method for the identification of high risk infants in clinics in the future, a large-scale study involving a mixed ethnically diverse population is much needed. PMID- 17119994 TI - Functional outcome and prognostic factors in 304 eyes managed by retinectomy. AB - BACKGROUND: This study was undertaken to relate the anatomic and functional results of patients who underwent retinectomy for complex retinal detachment (RD) to preoperative prognostic variables. METHODS: Three hundred and four eyes of 302 patients whose surgery involved retinectomy were included in the analysis. All eyes had established proliferative vitreoretinopathy (PVR grade C). The main outcome measures were (1) postoperative visual acuity of 6/24 or better, (2) status of the retina at the end of follow-up, and (3) incidence of hypotony whilst under review. RESULTS: PVR was secondary to rhegmatogenous RD in 237 eyes (78%), posterior trauma in 51 eyes (16.8%), tractional RD in vasoproliferative vasculitides in 12 eyes (4%), acute retinal necrosis in 2 eyes and endophthalmitis in 2 eyes. Complete reattachment rate after one operation was 51%, with final complete reattachment success rate of 72%. The visual acuity improved in 138 eyes (45%), remained the same in 73 eyes (24%) and became worse in 89 cases (29%). Postoperative visual acuity of 6/24 or better was significantly associated with preoperative vision, the duration of silicone oil tamponade, silicone oil removal and retinectomy size. There was also some evidence of association between visual outcome and the number of clock hours of retinal detachment. Final retinal attachment was significantly associated with silicone oil removal and preoperative vision, and final hypotony was significantly associated with silicone oil removal. The incidence of sympathetic ophthalmia in our study was 0.09% (one case). CONCLUSIONS: Good functional outcome is possible following retinectomy surgery despite advanced pathology and often multiple surgical procedures. Retinal redetachment as a result of reproliferation and hypotony appear to be the main reasons for anatomical and functional failure. The clinical features we have identified as good indicators for improved final visual acuity such as shorter tamponade duration, removal of silicone oil, smaller retinectomy size, fewer previous operations and better preoperative vision are surrogate markers of less advanced PVR and should prompt retinal surgeons to consider retinectomy at an earlier stage in the process of PVR development. Clinicians should be aware of the small risk of sympathetic ophthalmia from complex retinal surgery. PMID- 17119995 TI - Influence of macular pigment and melanin on incident early AMD in a white population. AB - BACKGROUND: The protective effect of macular pigment (MP) and melanin against age related macular degeneration (AMD) is still controversial from cross-sectional studies. In an attempt to clarify this issue, we performed a population-based longitudinal study. METHODS: MP optical density (MPOD) and melanin optical density (MOD) data were collected during the second follow-up phase of the Rotterdam Study in 1999 in a random subset of 435 participants. Data from 419 participants (98% white) was available for analysis. AMD diagnosis was based on standardized fundus photographs according to the International Classification System, and AMD cases were subdivided into five mutually exclusive stages. In the three follow-up phases, incident AMD (iAMD), defined as absence of any AMD at baseline and the presence of stage 2 or higher at follow-up, was determined. We used Cox regression analysis to study the effect of an assumedly stable MPOD and MOD on early iAMD. RESULTS: During a mean follow-up of 9.82 years, 13 male and 17 female participants developed early iAMD and two male participants late iAMD. Because only two participants developed late iAMD, we had to restrict our analyses to early iAMD. Cox regression analysis adjusted for age and gender showed no significant effect of MPOD [hazard ratio (HR) 0.37; 95% confidence interval (CI) 0.04, 3.57] and MOD (HR 0.56; 95% CI 0.09, 3.60) on early iAMD. Additional adjustment for smoking did not change these associations. CONCLUSIONS: This unique but quantitatively limited material leads to the conclusion that no major protective effect of MPOD or MOD was seen on early iAMD. PMID- 17119996 TI - Survival after primary enucleation for choroidal melanoma: changes induced by the introduction of conservative therapies. AB - BACKGROUND: Most uveal melanomas are currently treated by eye-preserving radiotherapies. However, for melanomas of the largest size or with initial complications, enucleation remains the reference treatment. Enucleation is called primary when it is proposed as the only local treatment option for a melanoma. There is very little literature on the use of primary enucleation after the introduction of conservative treatments. Our main goal was to evaluate the survival of melanoma patients treated by primary enucleation since the introduction of proton-beam therapy in France in 1991. METHODS: All melanoma patients undergoing primary enucleation in our department between 1991 and 2002 were included in this retrospective study. The 5-year melanoma-specific survival rate was calculated using the Kaplan-Meier method. The multivariate prognostic analysis was performed using the Cox proportional hazards model. RESULTS: Forty patients, representing 8% of all patients with choroidal uveal melanoma diagnosed and followed up in our department during an 11-year period, were included in the study. No patient was lost to follow-up. The 5-year melanoma-specific survival rate was 31.45% (SE: 7.8) after primary enucleation. Significant prognosis factors in the multivariate analysis were: tumor thickness > 12 mm (p = 0.03), anterior margin of the tumor involving the iris (p = 0.018), and presence of epithelioid cells (p = 0.02). CONCLUSIONS: The very low survival rate reported reflects the evolution of primary enucleation, which is currently indicated only for melanomas with the worst prognosis. The knowledge of current post-enucleation survival rates represents an essential achievement for both correct assessment of conservative therapies and patient counseling. PMID- 17119997 TI - A pilot study on slit lamp-adapted optical coherence tomography imaging of trabeculectomy filtering blebs. AB - BACKGROUND: Our study aims to identify anatomical characteristics of glaucoma filtering blebs by means of slit lamp-adapted optical coherence tomography (SL OCT) and to identify new parameters for the functional prognosis of the filter in the early post-operative period. METHODS: Patients with primary open-angle glaucoma, aged 18 years and older, scheduled for primary trabeculectomy at the Department of Ophthalmology, Radboud University Nijmegen Medical Centre, were considered for our study. All patients underwent standardized trabeculectomy with intra-operative application of mitomycin C. The filtering blebs were evaluated clinically and with SL-OCT on day 1 and 1, 2, 4 and 12 weeks following surgery. The resulting data were analysed and weighed against surgical success. To better understand the SL-OCT data a small comparative histologic study was performed. RESULTS: The study included 20 eyes of 20 patients. After completion of our study, 15 eyes of 15 patients (mean age+/-SD 67 +/- 16 years) were eligible for data analysis and 5 eyes missed at least one follow-up visit. Filtering surgery was considered successful (intraocular pressure < or = 21 mmHg without antiglaucomatous medication) in 11 of 15 eyes. SL-OCT frequently demonstrated multiple hypo-reflective layers within Tenon's capsule ("striping" phenomenon) in the first post-operative week. Presumably, these layers corresponded with drainage channels in the histological specimen. These channels were present in functional filters but not in the failures. In addition, the visualisation of the sclera below the filtering zone was better defined in failures compared with successful filtering blebs ("shading" phenomenon). We observed no differences in the volume and clinical aspect of the blebs in the successful group compared with the unsuccessful group. CONCLUSIONS: Successful filtering blebs show characteristic optical properties on SL-OCT. These phenomena suggest a diffusely enhanced fluid content and the presence of intra-bleb drainage channels in functional filtering blebs. PMID- 17119998 TI - The efficacy of acrylic acid grafting and arginine-glycine-aspartic acid peptide immobilization on fibrovascular ingrowth into porous polyethylene implants in rabbits. AB - PURPOSE: To determine the effects of acrylic acid (AA) grafting by argon plasma treatment and of immobilization of arginine-glycine-aspartic acid (RGD) peptides on fibrovascular ingrowth rate into high-density porous polyethylene (HPPE) anophthalmic orbital implants. MATERIALS AND METHODS: Sixty rabbits were divided into three groups, with 20 rabbits in each group: (1) control group, rabbits implanted with unmodified HPPE; (2) PAA group, rabbits implanted with HPPE grafted with poly(AA) by argon plasma treatment; (3) RGD group, rabbits implanted with HPPE grafted with AA by argon plasma treatment and subsequently immobilized with RGD peptide. An HPPE spherical implant was put in the abdominal muscles of rabbit. After implantation for 4 weeks, the retrieved implants were sectioned and stained with hematoxylin and eosin (H&E). Blood vessels were counted using CD-31 immunostaining. Cross-sectional areas of fibrovascular ingrowth, blood vessel densities, and host inflammatory response scores were determined for all three groups. RESULTS: The mean cross-sectional areas of fibrovascularization at 2 and 3 weeks after implantation were the greatest in the RGD group, followed by the PAA group. While minimal fibrovascular ingrowths were noted in all implants at 1 week, all the implants showed nearly complete ingrowth at 4 weeks. Blood vessel densities were the highest in the RGD group, followed by the PAA group at 2, 3, and 4 weeks. The mean inflammation scores of the PAA and RGD groups were less than that of the control group. CONCLUSION: Fibrovascularization into HPPE implants was enhanced by surface grafting of AA and further improved by immobilizing RGD peptides onto the grafted AA surfaces. The inflammatory reactions were mild by either technique of surface modification. PMID- 17119999 TI - Comment re: multicenter study of the European Assessment Group for Lysis in the Eye (EAGLE) for the treatment of central retinal artery occlusion: design issues and implications. PMID- 17120000 TI - Comparison of simultaneous readings of intraocular pressure in rabbits using Perkins handheld, Tono-Pen XL, and TonoVet tonometers. PMID- 17120001 TI - A case of cutaneous melanoma metastatic to the vitreous cavity: possible pathomechanism and review of the literature. AB - BACKGROUND: Isolated vitreous metastases are extremely rare and the pathogenesis of metastasis is still unclear. Here we present the detailed description of the disease progression in a 68-year-old patient with vitreous seeding of a metastatic cutaneous melanoma beginning at a very early stage. METHODS: Interventional case report and review of the literature. RESULTS: The initial retrohyaloidal metastatic lesion was identified adjacent to a small epiretinal hemorrhage. As the disease progressed golden brown spherules appeared in the posterior vitreous emanating from the area of the lesion. Further progression led to a dense metastatic infiltration of the entire vitreous cavity and a decline of the visual acuity to 20/1200. Diagnostic and therapeutic pars plana vitrectomy was performed to confirm the diagnosis and preserve the eye and useful vision. CONCLUSIONS: For the first time the formation of vitreous metastases derived from cutaneous melanoma was carefully studied beginning at a very early stage. This made it possible to analyze the rare mechanism of vitreous metastasis, which has not been conclusively known till now. The features of metastatic cutaneous melanoma to the vitreous are discussed in context of a review of the literature that resulted from the study of 17 patients with 22 affected eyes. PMID- 17120002 TI - Drug delivery to the eye through systematic circulation following subconjonctival injection. PMID- 17120003 TI - Bilateral canaliculitis following SmartPLUG insertion for dry eye syndrome post LASIK surgery. AB - BACKGROUND: Dry eyes are a common symptom following LASIK corneal refractive surgery. Treatments include topical lubricants to supplement the tear film, and punctal occlusion to reduce tear outflow. Canaliculitis is a recognised complication of punctal plugs, but has not previously been described following insertion of newer generation semi-permanent intra-canalicular plugs, such as the SmartPLUG. METHODS: Case report. RESULTS: We describe a 60-year-old female who underwent bilateral LASIK surgery leading to aggravation of her dry eye syndrome. She was managed with the insertion of semi-permanent intra-canalicular moldable silicone SmartPLUGs. She subsequently developed bilateral canaliculitis requiring bilateral canaliculotomy. CONCLUSIONS: To the best of our knowledge, this is the first report of bilateral canaliculitis following intra-canalicular SmartPLUG insertion. This case illustrates that punctal occlusion with the newer generation intra-canalicular plugs carries a risk of canaliculitis and that irrigation is not always effective in removing these devices. PMID- 17120004 TI - Ocular penetration of caspofungin in a rabbit uveitis model. AB - BACKGROUND: Little is known about the ocular penetration of echinocandin antifungals. We studied the ocular distribution of systemically administered caspofungin in a rabbit uveitis model. METHODS: Caspofungin (1 mg/kg per day) was given intravenously to rabbits as a single dose or as repeated daily doses on 7 days starting 24 h after induction of unilateral uveitis by intravitreal endotoxin injection. Caspofungin concentrations were determined by high performance liquid chromatography in the cornea, aqueous humor, vitreous humor, and serum 4, 8, 16, and 24 h after administration of a single dose and 24 h after the last of seven doses. RESULTS: The mean caspofungin concentration in the aqueous of the inflamed eye 4 and 8 h after single-dose administration was 1.30 +/- 0.39 microg/ml and 1.12 +/- 0.34 microg/ml, respectively. Drug concentrations decreased to 0.24 +/- 0.09 microg/ml at 16 h and 0.26 +/- 0.14 microg/ml at 24 h. In the vitreous of inflamed eyes drug levels were undetectable at all time points. No drug was found in the aqueous of inflamed eyes 24 h after the last of seven repeated doses, and the vitreous only contained trace amounts. In the corneas of inflamed eyes concentrations reached 1.64 +/- 0.48 microg/g at 4 h, peaked at 2.16 +/- 1.14 microg/g at 8 h, and declined to 1.87 +/- 0.52 microg/g and 1.49 +/- 0.48 microg/g at 16 and 24 h, respectively. After repeated dosing, corneal concentrations of caspofungin were 0.8 and 1.0 microg/g and below the limit of detection in two of four animals. In non-inflamed eyes no drug was detectable in the aqueous and vitreous humor, and the corneas at any time point. CONCLUSIONS: In our model, caspofungin reached therapeutically relevant levels in the aqueous and cornea but not in the vitreous humor of inflamed eyes. Intraocular drug deposition was critically dependent on a disrupted blood-eye barrier. These findings suggest a limited role for caspofungin in the treatment of fungal endophthalmitis. PMID- 17120005 TI - Reelin expression is upregulated following ocular tissue injury. AB - PURPOSE: Reelin is important in the guidance of neuronal stem cells in the central nervous system during normal development. We wished to determine whether reelin is expressed in the retina and cornea after injury. METHODS: Mice underwent laceration of their retina as well as corneal epithelial debridement. The mice were sacrificed at 3 days, and eyes were fixed and stained for reelin expression and reelin messenger ribonucleic acid (mRNA). RESULTS: In normal eyes, reelin was expressed only at very low levels in the ganglion cell layer of the retina and the endothelial cell layer of the cornea. In injured eyes, there was marked expression in reelin immunoreactivity in the retina and cornea. Reelin gene expression was seen in the retina and cornea. CONCLUSIONS: Reelin is expressed during normal retinogenesis. This study shows that reelin is also upregulated following injury to the retina and cornea. The expression of reelin following injury suggests that reelin may play an important role in regulating stem cell trafficking in neuronal and nonneuronal tissues following injury similar to its role in normal organogenesis. PMID- 17120007 TI - Acquired choroidal folds: a sign of idiopathic intracranial hypertension. AB - BACKGROUND: Choroidal folds may be caused by several underlying ocular and orbital diseases; they are classified as idiopathic when no obvious cause is found. The objective of this study was to determine whether acquired choroidal folds are associated with idiopathic intracranial hypertension. METHODS: In this observational case series, three patients with normal fundus examination later developed choroidal folds. They underwent complete ocular examination, B-scan ultrasonography, fluorescein angiography, optic coherent tomography (OCT) and magnetic resonance imaging. A neurological consultation, lumbar puncture and measurement of opening pressure of cerebrospinal fluid (CSF) were also obtained for all patients. RESULTS: Three patients with previously normal ocular fundus developed choroidal folds, and optic nerve subarachnoid space enlargement was seen on B-scan. Clinical and radiological evaluations identified elevation of intracranial pressure, and biochemical analysis of CSF was normal, suggesting a diagnosis of pseudotumour cerebri, which was later confirmed. The first patient developed choroidal folds in one eye first, and 4 years later in the fellow eye. The second patient presented with bilateral optic disk swelling secondary to intracranial hypertension. After treatment, which lowered CSF pressure, bilateral choroidal folds were identified by OCT, fluorescein angiography and ocular fundus photograph. The third patient also developed choroidal folds in one eye, but his fellow eye was difficult to evaluate due to a chorioretinitis scar on the macula. OCT identified choroidal folds in all three patients and in the second patient, it also revealed retinal nerve fiber layer damage. CONCLUSIONS: Acquired choroidal folds and optic nerve subarachnoid space enlargement may be signs of idiopathic intracranial pressure elevation. Nevertheless, this diagnosis should be confirmed by lumbar puncture. Comprehensive imaging studies should be performed to rule out expanding tumors. The lower CSF pressure in our first two patients suggests that choroidal folds or optic disk swelling may depend on the level of intracranial pressure and may be points in a continuum of clinical presentations. PMID- 17120008 TI - The long-term safety of donor eye for 180 degrees limbal transplantation. AB - BACKGROUND: Little information is currently available on the long-term safety of eyes following limbal tissue donation. We have therefore evaluated long-term refractive changes, corneal surface stability, and visual outcome following limbal tissue donation of 180 degrees with permanent amniotic membrane transplantation (AMT). METHODS: Four healthy eyes of four patients underwent limbal keratectomy involving half of the limbal circumference (half limbal) for the purpose of limbal donation with permanent AMT. The best corrected visual acuity (BCVA), astigmatic changes by manifest refraction, and corneal surface changes as assessed by slit-lamp examination and by impression cytology were evaluated. RESULTS: The mean follow-up period was 20.8 months (range: 19-24 months). BCVA remained unchanged in three eyes (75%), but eye one showed a BCVA decrease of two lines due to newly developed filamentary keratitis caused by Sjogren's syndrome. The average astigmatism change was an increase of 0.31 diopters. Conjunctival invasion toward the center beyond the edge of the keratectomy did not occur in any of the eyes, as assessed by impression cytology. Partial conjunctivalization within 2 h on keratectomy sites covered with amniotic membrane developed in three eyes (75%). CONCLUSIONS: Limbal donation of 180 degrees with permanent AMT seems to be a safe procedure, demonstrating stable astigmatism and an intact corneal surface for at least 1.5 years following the initial donation. PMID- 17120006 TI - Vascular risk factors in glaucoma: the results of a national survey. AB - BACKGROUND: The role of vascular risk factors in glaucoma is still being debated. To assess the importance of vascular risk factors in patients with primary open angle glaucoma (POAG), data from the medical history of 2,879 POAG patients and 973 age-matched controls were collected and analyzed. METHODS: DESIGN: observational survey. SETTING: 35 Italian academic centers. STUDY POPULATION: POAG patients and age-matched controls. In order to reduce bias consecutive patients were included. OBSERVATION PROCEDURES: data concerning vascular risk factors were collected for all patients with a detailed questionnaire. A complete ophthalmological examination with assessment of intraocular pressure (IOP), visual field, optic disc, and systemic blood pressure was performed. MAIN OUTCOME MEASURES: the ESH-ESC (European Society of Hypertension-European Society of Cardiology) guidelines were used to calculate the level of cardiovascular risk. Crude and adjusted estimates of the odds ratios (OR) were calculated for all cardiovascular risk factors in POAG and controls. RESULTS: The study included 2,879 POAG patients and 973 controls. POAG cases had a significantly higher systolic and diastolic blood pressure (p = 0.001) and systolic perfusion pressure (p = 0.02) as compared with controls. Also mean IOP was significantly higher in the POAG group (p = 0.01), while diastolic perfusion pressure was not significantly different in the two groups. Myopia was more prevalent in the POAG group (23 vs 18%, p = 0.005) as well as a positive family history for glaucoma (26 vs 12%, p = 0.004). POAG patients tended to have a higher cardiovascular risk than controls: 63% of glaucoma cases vs 55% of controls (OR: 1.38, p = 0.005) had a "high" or "very high" cardiovascular risk. CONCLUSIONS: The level of cardiovascular risk was significantly higher in glaucoma patients than in controls. PMID- 17120009 TI - Conjunctival mucin deficiency in complete androgen insensitivity syndrome (CAIS). AB - Sex steroid hormones are essential for a healthy ocular surface and the androgen receptor impairment found in patients with complete androgen insensitivity syndrome (CAIS) has been described to cause meibomian gland dysfunction and functional dry eye for lipid tear film layer instability. However, it has not been reported if the mucous layer is also affected. A 37-year-old CAIS patient with persistent symptoms of dry eye underwent ophthalmological examination and was evaluated for qualitative and quantitative tear function tests and conjunctival cytology. Samples obtained from the conjunctival epithelium were stained for histology and immunohistochemistry and compared with three age matched female controls. Western blot and relative real-time RT-PCR for MUC1 and MUC5AC were also performed on these samples. Immunohistochemistry, Western blot and relative real-time RT-PCR showed a decrease in the expression of MUC1 and MUC5AC in CAIS. Changes in the tear film mucous layer were accompanied by a reduction in the tear film break up time test. This is the first report describing mucous layer alteration associated with androgen receptor impairment. Decreased mucin levels contribute in explaining the tear film instability in CAIS and should be considered an additional cause of dry eye in sex steroid hormone pathology. PMID- 17120010 TI - The phenomenon of "sticky" silicone oil. AB - BACKGROUND: To investigate the reasons for difficulties removing silicone oil from the vitreous cavity due to putative adherence to the retina. METHODS: Gas chromatography-coupled mass spectroscopy of the headspace (GC/MS/HS) and gel permeation chromatography (GPC) were used to detect volatile compounds in silicone oil samples explanted from patients, qualitatively as well as quantitatively. Surface and interfacial tensions of the explanted samples were measured using the pendent-drop technique. To simulate the removal of silicone oil from the vitreous cavity, the contact between silicone oil and differently treated surfaces and various aspiration techniques were tested in vitro. RESULTS: The median concentration of perfluorodecalin in seven "sticky" samples was 2.4 times higher than in 14 non-sticky samples. In the sticky samples, the median surface tension of the aqueous phase was lower. The difficulty of aspirating silicone oil could be reproduced in vitro by reducing the surface tension of the aqueous environment of the silicone oil. CONCLUSION: The observed stickiness of silicone oil seems to be a matter of reduced surface tension of the surrounding aqueous material and/or contamination of silicone oil with perfluorocarbon liquid, which creates interruption of the material flow, giving the impression of adherence of the silicone oil to the retina. PMID- 17120011 TI - Histological and immunohistochemical characterisation of conjunctival graft vs host disease following haematopoietic stem cell transplantation. AB - BACKGROUND: Conjunctival graft vs host disease (cnGvHD) is a complication of haematopoietic stem cell transplantation, in most cases as part of systemic GvHD. Diagnostic biopsies are commonly collected from bulbar conjunctiva only. The aims of our study were to evaluate whether additional biopsies from the tarsal conjunctiva increase sensitivity upon histopathologic evaluation and to investigate the staining profile for common immunohistochemical markers in cnGvHD. We additionally propose an adaptive histological classification for cnGvHD analogous to Lerner's GvHD skin classification for predicting patient survival. METHODS: Formalin-fixed and paraffin-embedded conjunctival specimens from 23 post-mortem control eyes and 42 patients after haematopoietic stem cell transplantation (HSCT) were stained with haematoxylin and eosin (HE), periodic acid-Schiff (PAS) stain and with antibodies against CD1a, CD4, CD8, CD25, CD45RO, CD68, Fas ligand, TIA-1, HLA-DRalpha by means of immunohistochemistry. Cell counting took place in ten representative fields at 64.4 microm (length) x 21.2 microm (width). Multifactorial analysis of variance was performed to assess any influence of cnGvHD on the staining pattern for the immunohistochemical markers. Survival times were estimated by the Kaplan-Meier method. RESULTS: All 42 specimens and none of the controls were diagnosed as cnGvHD. The bulbar specimens were staged according to the modified Lerner classification: grade (G) I: 0; G II: 17 (tarsal with GII, 8); G III: 12 (tarsal with GIII: 1); G IV: 12 (tarsal with G or = 50). Antibodies were seen in 10 (20%) of 50 household dogs and in 23 (46%) of 50 farm dogs. There were significant statistical differences in seropositivity between these two groups (P = 0.005). The IFAT antibody titers were as follows: 1:50 in seven dogs, 1:100 in eight dogs, 1:200 in six dogs, 1:400 in seven dogs, 1:800 in three dogs, 1:1,600 in one dog, and 1:12,800 in one dog. There were no significant differences in seropositivity between males and females. The positive results were increasing with age, and positive results were significantly different in the age group of older than 2 years compared to the dogs of age group under 1 year (P = 0.000) and 1-2 years (P = 0.007). The results confirm the exposure of household and farm dogs to N. caninum in Tehran and the higher rate of exposure for the dogs of dairy and cattle farms around Tehran. PMID- 17120043 TI - Parasites of two abundant sympatric rodent species in relation to host phylogeny and ecology. AB - In the present study, two abundant sympatric rodent species (27 Apodemus flavicollis and 33 A. sylvaticus) were studied for their endo- and ectoparasite fauna. The rodents were trapped in Dormagen, a city in North Rhine-Westphalia, Germany. A total of 20 different parasites species were identified, 13 endoparasite (2 Digenea, 5 Cestoda and 7 Nematoda) and 7 ectoparasite (5 Insecta and 2 Arachnida) species. Thirteen parasite species were found inhabiting both rodent species. The predominant endoparasite species in both rodents was the nematode Pelodera strongyloides, followed by the nematode Heligmosomoides polygyrus and a Syphacia species. The flea Ctenophthalmus agyrtes was the dominant ectoparasite in both rodent species. A. flavicollis usually carried 1-7 ecto-/endoparasite species (mean 4.0), whereas A. sylvaticus were mostly infested with 1-9 (mean 4.4). The parasite diversity of A. flavicollis (H' = 0.268, J = 0.097) was marginal lower in comparison to A. sylvaticus (H' = 0.319, J = 0.110). The two rodent species examined show remarkable similarities in the composition of their endo- and ectoparasite fauna being directly related to their similar pattern of living in the investigated area. PMID- 17120044 TI - Efficacy of oxfendazole and fenbendazole against tortoise (Testudo hermanni) oxyurids. AB - Thirty-six tortoises (Testudo hermanni) with naturally acquired oxyurids infections were used to assess the anthelmintic efficacy of oxfendazole (Dolthene; Merial) and fenbendazole (Panacur; Hoechst Roussel Vet). Animals were randomly assigned to three groups (A, B, and C) based on sex and weight. Animals in group A (seven males and six females) were orally treated with oxfendazole at dose rate of 66 mg/kg, group B animals (nine males and eight females) were orally treated with fenbendazole at dose rate of 100 mg/kg, and group C animals (three males and three females) were not treated and served as controls. All animals were individually stabled in plexiglas boxes under controlled conditions of temperature, humidity, and light beginning 7 days pretreatment and continuing for the duration of the trial. Individual tortoises feces were examined daily by the McMaster technique and drugs efficacy was assessed by the fecal eggs count reduction (FECR) test. Both drugs showed 100% of FECR. However, oxfendazole reached this level 12 days after treatment, whereas 31 days after treatment were necessary to obtain the same stable result with fenbendazole. The two drugs were well tolerated by all the animals and no adverse reactions were observed after treatment. PMID- 17120045 TI - New data on Microphallus breviatus Deblock & Maillard, 1975 (Microphallidae: Digenea) with emphasis on the evolution of dixenous life cycles of microphallids. AB - Infection by intramolluscan stages of Microphallus breviatus Deblock & Maillard, 1975 are common in Hydrobia ventrosa mudsnails in Iceland. Cercariae encyst inside the daughter sporocysts and develop there into metacercariae that become infective for the definitive hosts which are probably charadriiform waders. The adult stage was obtained in 1-day-old chicks that were experimentally infected with metacercariae from naturally infected hydrobians. New data are presented on the morphology and biology of the cercariae, and the adult is described for the first time. Comparisons are made between M. breviatus and closely related species. Differential diagnosis of M. breviatus is given. The morphological specializations in larvae of the hermaphroditic generation of the microphallids accompanying transition from trixenous life cycles to dixenous ones are considered, and the applicability of the term "life-cycle truncation" to microphallids with dixenous life cycles is discussed. Also, reasons for the broad distribution of dixenous life cycles within the family Microphallidae are reviewed. PMID- 17120046 TI - Homozygosity mapping in consanguineous families reveals extreme heterogeneity of non-syndromic autosomal recessive mental retardation and identifies 8 novel gene loci. AB - Autosomal recessive gene defects are arguably the most important, but least studied genetic causes of severe cognitive dysfunction. Homozygosity mapping in 78 consanguineous Iranian families with nonsyndromic autosomal recessive mental retardation (NS-ARMR) has enabled us to determine the chromosomal localization of at least 8 novel gene loci for this condition. Our data suggest that in the Iranian population NS-ARMR is very heterogeneous, and they argue against the existence of frequent gene defects that account for more than a few percent of the cases. PMID- 17120047 TI - A novel polymorphism associated with lactose tolerance in Africa: multiple causes for lactase persistence? AB - Persistence or non-persistence of lactase expression into adult life is a polymorphic trait that has been attributed to a single nucleotide polymorphism (C 13910T) in an enhancer element 13.9 kb upstream of the lactase gene (LCT). The 13910*T allele occurs at very high frequency in northern Europeans as part of a very long haplotype (known as A), and promotes binding of the transcription factor Oct-1. However, -13910*T is at very low frequency in many African milk drinking pastoralist groups where lactase persistence phenotype has been reported at high frequency. We report here for the first time, a cohort study of lactose digester and non-digester Sudanese volunteers and show there is no association of -13910*T or the A haplotype with lactase persistence. We support this finding with new genotype/phenotype frequency comparisons in pastoralist groups of eastern African and Middle Eastern origin. Resequencing revealed three new SNPs in close proximity to -13910*T, two of which are within the Oct-1 binding site. The most frequent of these (-13915*G) is associated with lactose tolerance in the cohort study, providing evidence for a cis-acting effect. Despite its location, 13915*G abolishes, rather than enhances Oct-1 binding, indicating that this particular interaction is unlikely to be involved in lactase persistence. This study reveals the complexity of this phenotypic polymorphism and highlights the limitations of C-13910T as a diagnostic test for lactase persistence status, at least for people with non-European ancestry. PMID- 17120048 TI - Fine-mapping the putative chromosome 17q21-22 prostate cancer susceptibility gene to a 10 cM region based on linkage analysis. AB - Prostate cancer linkage studies have suggested the existence of a prostate cancer susceptibility gene on chromosome 17q21-22. We now report the results of an extended linkage analysis including 95 new multiplex prostate cancer families and 9 additional microsatellite markers resulting in a maximum LOD score of 2.99 at approximately 81-82 cM for all 453 pedigrees. Results from these 95 new pedigrees provide additional support for a chromosome 17q21-22 prostate cancer susceptibility gene. Inclusion of the 9 additional markers significantly reduced the size of the candidate region, as defined using a 1-LOD support interval, especially when focusing analyses on subsets of pedigrees with four or more confirmed affecteds or average age of diagnosis less than or equal to 65 years. A novel subset analysis of only those families (n = 147) that had four or more prostate cancer cases and an average age of prostate cancer diagnosis < or = 65 years results in a maximum LOD score of 5.49 at 78 cM with a 1-LOD support interval of 10 cM. This large set of pedigrees with four more prostate cancer cases characterized by early-onset disease will serve as a useful resource for identifying the putative 17q21-22 prostate cancer susceptibility gene. PMID- 17120050 TI - Distributional characteristics of lymphatic vessels in normal human nasal mucosa and sinus mucosa. AB - An immunohistochemical staining technique with the D2-40 antibody was undertaken to examine the functional and morphological features of lymphatic networks in tissue sections and whole-mount preparations of normal nasal mucosa and ethmoid sinus mucosa. In normal nasal mucosa, most lymphatic vessels were found in the superficial mucosa beneath the epithelial layer. Some of these vessels were dilated, whereas others were compressed and had a slit-like lumen. Whole-mount preparations revealed the extent of lymphatic vessels in normal ethmoid sinus mucosa. A network of lymphatic vessels was mainly found in the subepithelial layer, where lymphatic vessels represented rich networks, possessing antler-like branches and typical blind ends. However, these lymphatic networks were not arranged in the form of lymphangion chains, with each lymphangion consisting of a contractile compartment and valve. Thus, recognition of the distinctive features of the lymphatic network in normal nasal and sinus mucosa might aid investigations of lymphatic involvement in sinonasal diseases, such as rhinitis, sinusitis, and malignancy. PMID- 17120049 TI - Contributions of the DAT1 and DRD2 genes to serious and violent delinquency among adolescents and young adults. AB - As far as we know, this is the first national study that reports compelling evidence for the main effects of genetic variants on serious and violent delinquency among adolescents and young adults. This study investigated the association between the self-reported serious and violent delinquency and the TaqI polymorphism in the DRD2 gene and the 40-bp VNTR in the DAT1 gene. The study was based on a cohort of more than 2,500 adolescents and young adults in the National Longitudinal Study of Adolescent Health in the United States. The trajectories of serious delinquency for the DAT1*10R/10R and DAT1*10R/9R genotypes are about twice as high as that for the DAT1*9R/9R genotype (LR test, P = 0.018, 2 df). For DRD2, the trajectory of serious delinquency for the heterozygotes (A1/A2) is about 20% higher than the A2/A2 genotype and about twice as high as the A1/A1 genotype, a phenomenon sometimes described as heterosis (LR test, P = 0.005, 2 df). The findings on violent delinquency closely resemble those on serious delinquency. The trajectories of violent delinquency for the DAT1*10R/9R and DAT1*10R/10R genotype are again about twice as high as that for DAT1*9R/9R (LR test, P = 0.021, 2 df). The two homozygotes of DRD2*A1/A1 and DRD2*A2/A2 scored lower (LR test, P = 0.0016, 2 df) than the heterozygotes. The findings in the models that consider DAT1 and DRD2 jointly (serious delinquency P = 0.0016, 4 df; violent delinquency P = 0.0006, 4 df) are essentially the same as those in the single-gene models, suggesting the absence of a significant correlation between the two genetic variants. These results only apply to males. Neither variant is associated with delinquency among females. PMID- 17120051 TI - Identification of key tissue type for antler regeneration through pedicle periosteum deletion. AB - Epimorphic regeneration is the "holy grail" of regenerative medicine. Research aimed at investigating the various models of epimorphic regeneration is essential if a fundamental understanding of the factors underpinning this process are to be established. Deer antlers are the only mammalian appendages that are subject to an annual cycle of epimorphic regeneration. In our previous studies, we have reported that histogenesis of antler regeneration relies on cells resident within the pedicle periosteum (PP). The present study elaborates this finding by means of functional studies involving the deletion of PP. Four yearling and four 2-year old stags were selected for total PP deletion or partial PP deletion experiments. Of the animals in the total PP deletion group, one showed no signs of antler regeneration throughout the antler growth season. Two showed substantial and one showed marginal delays in antler regeneration (at 34, 20 and 7 days, respectively) compared with the corresponding sham-operated sides. Histological investigation revealed that the delayed antlers were derived from regenerated PP. Unexpectedly, the regenerative capacity of the antler from the total periosteum deleted pedicles depended on antler length at surgery. Of the four deer that had partial PP deletion, two regenerated antlers exclusively from the left-over PP on the pedicle shafts in the absence of participation from the pedicle bone proper. The combined results from the PP deletion experiments convincingly demonstrate that the cells of the PP are responsible for antler regeneration. PMID- 17120052 TI - Histological changes in intestine of Atlantic salmon (Salmo salar L.) following in vitro exposure to pathogenic and probiotic bacterial strains. AB - Furunculosis and vibriosis are diseases that cause severe economic losses in the fish-farming industry. The foregut of the Atlantic salmon (Salmo salar L.) was exposed in vitro to two fish pathogens, Aeromonas salmonicida (causative agent of furunculosis) and Vibrio anguillarum (causative agent of vibriosis), and to one probiotic strain, Carnobacterium divergens, at 6 x 10(4) or 6 x 10(6) viable bacteria per milliliter. Histological changes following bacterial exposure were assessed by light and electron microscopy. Control samples (foregut exposed to Ringer's solution only) and samples exposed only to C. divergens had a similar appearance to intact intestinal mucosal epithelium, with no signs of damage. However, exposure of the foregut to the pathogenic bacteria resulted in damaged epithelial cells, cell debris in the lumen, and disorganization of the microvilli. Co-incubation of the foregut with a pathogen and C. divergens did not reverse the damaging effects caused by the pathogen, although these were alleviated when probiotic bacteria were used. Based on these results, we suggest that the probiotic bacterium, C. divergens, is able to prevent, to some extent, pathogen-induced damage in the Atlantic salmon foregut. PMID- 17120053 TI - Secretory granules of hypophyseal and pancreatic endocrine cells contain proteins of the neuronal postsynaptic density. AB - The PDZ domain-containing protein Shank is a master scaffolding protein of the neuronal postsynaptic density and directly or indirectly links neurotransmitter receptors and cell adhesion molecules to the actin-based cytoskeleton. ProSAP/Shank proteins have recently also been detected in several non-neuronal cells in which they are mostly concentrated in the apical subplasmalemmal cytoplasm. In contrast, we have previously reported a more widespread cytoplasmic immunostaining pattern for the ProSAP1/Shank2 protein in endocrine cells at the light-microscopic level. Therefore, in the present study, we have determined the ultrastructural localization of ProSAP1/Shank2 and the ProSAP/Shank-interacting proteins ProSAPiP1 and IRSp53 in pancreatic islet and adenohypophyseal cells by using immunogold staining techniques. Dense immunolabeling of secretory granules including the granule core in cells such as hypophyseal somatotrophs and pancreatic B-cells indicates the unexpected presence of ProSAP/Shank and ProSAP/Shank-interacting proteins in the hormone-storing compartment of endocrine cells. Thus, ProSAP/Shank and certain ProSAP/Shank-interacting proteins exhibit distinct subcellular localizations in the different cell types, raising the possibility that the function of ProSAP/Shank proteins is more diverse than has been envisaged to date. PMID- 17120054 TI - Nature of Cx30-containing channels in the adult mouse mammary gland. AB - Oligonucleotide microarray analysis uniquely shows that several members of the connexin family of gap junction proteins are expressed by the epithelium during mouse mammary gland development. Connexin 26 (Cx26) is present throughout pregnancy and lactation, is then undetectable shortly after weaning, but reappears during involution. Additionally, Cx30 is abundant in late-pregnant and early lactating gland epithelium. From mid-pregnancy into early lactation, Cx26 and Cx30 co-localize in junctional plaques between epithelial cells, forming hemichannels of mixed connexin content. Microarray analysis also shows Cx32 is developmentally restricted to parturition, suggesting that specific modification of gap junction channel composition and/or intercellular communication pathways occurs at parturition. Specifically, heteromeric channels of all pairwise combinations are formed when these connexins are expressed within the same cells. Of these hemichannels, Cx26/Cx32 pores are increasingly sensitive to closure by taurine (an osmolyte implicated in milk protein synthesis) with increasing Cx26 content. In contrast, physiological taurine concentrations have no effect on Cx26/Cx30 and Cx30/Cx32 channel activity. Such changes in connexin expression and channel composition and their chemical modulation are discussed in relation to the various stages of mammary gland development in the adult mouse. PMID- 17120055 TI - Variation in multiple traits of vegetative and reproductive seagrass tissues influences plant-herbivore interactions. AB - Plant-herbivore interactions have strong ecological and evolutionary consequences, but have been traditionally overlooked in marine higher plants. Despite recent advances in seagrass ecology that highlight the importance of herbivory, the mechanisms that regulate the feeding behaviour of seagrass consumers remain largely unknown. Herbivores have been shown to reduce the sexual reproductive success of seagrasses through direct consumption of inflorescences and seeds, but we know little about intraspecific variation in susceptibility to grazing of different seagrass tissues. We contrasted the relative palatability of reproductive and vegetative tissues of the temperate seagrass Posidonia oceanica in the field, and we assessed the feeding preferences among these tissues of the main consumers of the plant, the fish Sarpa salpa and the urchin Paracentrotus lividus. Moreover, we identified the plant traits that explained the observed feeding behaviour. We provide strong evidence for herbivore selectivity among seagrass tissues. In the field, 70-90% of inflorescences were damaged by herbivores compared to 3-60% of leaves of similar age. In feeding assays, the urchin P. lividus showed over a twofold preference for reproductive tissue at various stages of development. By contrast, we detected no feeding activity on either leaves or inflorescences from the fish S. salpa, which is known to migrate to deeper waters soon after flowering starts and during the period of fruit maturation. Despite being the preferred food of urchins, inflorescences were chemically defended, had higher levels of phenolics and lower nutrient and calorific content than leaves. We experimentally demonstrated that leaf structural defences are the primary factor in determining urchin feeding preferences. Removal of plant structure results in a drastic shift in urchin selectivity towards the most nutritious and less chemically defended leaf tissue, indicating that multiple mechanisms of defence to herbivory may coexist in seagrasses. PMID- 17120056 TI - Landscape context outweighs local habitat quality in its effects on herbivore dispersal and distribution. AB - Past studies with spatially structured herbivore populations have emphasized the primacy of intrinsic factors (e.g., patch quality), patch geometry (e.g., patch size and isolation), and more recently landscape context (e.g., matrix composition) in affecting local population abundance and dispersal rate. However, few studies have examined the relative importance of each factor, or how they might interact to affect herbivore abundance or dispersal. Here, we performed a factorial field experiment to examine the independent and interactive effects of patch quality (plant biomass, leaf protein, leaf phenolics) and matrix composition [mudflat or non-host grass (Bromus inermis)] on planthopper (Prokelisia crocea) emigration from host-plant patches (prairie cordgrass, Spartina pectinata). In addition, a field survey was conducted to examine the relative importance of patch quality, geography, and matrix composition on planthopper occupancy and density. In the experiment, we found that rates of emigration from low and intermediate quality patches were, on average, 21% percent higher for patches embedded in brome than mudflat. In contrast, the emigration rate was unaffected by matrix composition in nutrient-rich patches. Within matrix types, plant quality had little effect on emigration. In the survey, planthopper density and the patch occupancy rate of planthoppers increased nonadditively with increasing patch size and the percentage of the surrounding matrix composed of mudflat. This study suggests that landscape-level factors, such as the matrix, may be more important than factors intrinsic to the patches. PMID- 17120057 TI - Colonization of ephemeral detrital patches by vagile macroinvertebrates in a brackish lake: a body size-related process? AB - The mechanisms regulating the build-up of invertebrate assemblages on ephemeral detritus patches are still poorly understood. Here, the daily colonization of decaying reed leaves by vagile macroinvertebrates was monitored in an brackish lake in Italy. The highly variable abundance patterns of dominant taxa were analysed by spectral and geostatistical techniques to test for nonrandomness and to further determine whether they were related to body size. Comparisons between two contrasting sites allowed an assessment of the generality of our observations. At both sites, the macroinvertebrate assemblage was dominated by three detritivorous taxa, i.e. the isopod Lekanesphaera monodi, the amphipod Microdeutopus gryllotalpa and the polychaete Neanthes caudata. Overall, their abundance patterns were characterised by short-term fluctuations of a nonrandom, autocorrelated nature. In addition, a significant covariation was observed between the average body mass of each taxon and the complexity of the respective abundance pattern, expressed by the fractal dimension D. The covariation was observed at both study sites, notwithstanding the diverging outcomes of bivariate pattern comparisons for similar-sized taxa. Our findings indicate that the size of macroinvertebrates is strongly related to the short-term dynamics of their abundance patterns on reed detritus, suggesting that the interaction between vagile consumers and ephemeral resource patches might be influenced by individual energetics. The implications of size-related constraints for the coexistence of species on decaying detrital patches are discussed. PMID- 17120058 TI - Climate-driven warming during spring destabilises a Daphnia population: a mechanistic food web approach. AB - Temperature-driven changes in interactions between populations are crucial to the estimation of the impact of global warming on aquatic food webs. We analysed inter-annual variability in two data sets from Bautzen reservoir, Germany. In a long-term data set (1981-1999) we examined the pelagic phenology of Daphnia galeata, a keystone species, the invertebrate predator Leptodora kindtii, phytoplankton and Secchi depth in relation to water temperature and the North Atlantic Oscillation index. In a short-term data set (1995-1998) we examined food web relations, particularly the consumption of D. galeata by young-of-the-year (YOY) percids and L. kindtii and rates of population change of D. galeata (abundance, recruitment pattern and non-consumptive mortality). The start of the clear-water stage (CWS) was correlated with winter temperatures. It started 5.8 days earlier per degree warming after warm winters (mean January-March temperature>or=2.5 degrees C) compared to cold winters (mean temperature<2.5 degrees C). However, the end of the CWS remained relatively constant. Predation by L. kindtii and YOY percids on D. galeata started distinctly earlier, i.e. by 13.0 and 6.5 days per degree warming, respectively, in years when the average May temperature was high (>or=14 degrees C) compared to years when it was low (<14 degrees C). Significant reductions of Daphnia abundance in midsummer occurred only in years in which the mean May temperature exceeded 14 degrees C. This temperature regime resulted in a match of over-exploitation of food resources by Daphnia during the CWS and strong predation by YOY percids and L. kindtii. Consumptive mortality increased at higher rates with a rise in temperature than net recruitment, resulting in lower Daphnia densities at the end of the CWS. Our data suggest that even low warming by 1.7 degrees C during a short, but critical seasonal period, resulting in the coincidence of two or more factors adversely affecting a keystone species, such as Daphnia, may induce changes in whole lake food webs and thus alter entire ecosystems. PMID- 17120059 TI - Effects of the internal recycling rate on biological nutrient removal and microbial community structure in a sequential anoxic/anaerobic membrane bioreactor. AB - This study investigated the effects of the internal recycling rate on nutrients removal in a sequential anoxic/anaerobic membrane bioreactor (SAM). Microbial community structure in sludge from the SAM was studied using quinone profile method. Above 98% COD, 68% nitrogen, and 55% phosphorus removal efficiencies were achieved when the internal recycling rate was 2.5 times influent flow. At that rate, the optimum specific nitrate loading rate and COD/NO(3)-N ratio were found to be 2.24 mgNO(3)-N g(-1) MLSS h(-1) and 9.13, respectively. Batch tests demonstrated that anoxic condition suppressed phosphorus release, and that denitrification was also influenced by initial substrate concentration. Denitrification appeared to have some priority over phosphorus release for substrate uptake. Microbial community analysis revealed a predominance of the subclass beta-Proteobacteria. Furthermore, it was found that Rhodocyclus-related bacteria were efficient at phosphorus removal than Actinobacteria. PMID- 17120061 TI - Validation of child serum creatinine-based prediction equations for glomerular filtration rate. AB - Equations for estimating glomerular filtration rate (GFR) are vital in caring for patients with renal disease and the current standard, the Schwartz formula, lacks precision. We evaluated several child serum creatinine-based GFR prediction equations. Subjects aged 2-21 years who underwent iothalamate GFR (IoGFR) testing between 1999 and 2004 were studied retrospectively. GFR was estimated using: (1) Schwartz formula (SchwartzGFR), using a local k value; (2) Schwartz model (SchwartzMod) using regression-derived coefficients; (3) Leger GFR (LegerGFR) using original coefficients; and (4) Leger model (LegerMod) using regression derived coefficients. Bias, precision, and diagnostic characteristics were evaluated. There were 195 subjects [61% male; mean (SD) age 12.4 (4.5) years; mean (SD) IoGFR 78.9 (33.4) ml/min per 1.73 m(2)]. Only the LegerGFR overestimated IoGFR (5.5 ml/min per 1.73 m(2)). Precision for all formulae was poor (95% limits of agreement approximately -40 to 40 ml/min per 1.73 m(2)), but >or=72% of estimates were within 30% of IoGFR. Sensitivities for detecting IoGFR <30 and 90 ml/min per 1.73 m(2) were highest using the SchwartzGFR (80%) and SchwartzMod (90%), respectively. The LegerGFR was most specific. Using local coefficients, the Schwartz and Leger models were imprecise estimates of GFR, but the Schwartz model was most unbiased and sensitive. Future research should derive more precise equations for GFR in children. PMID- 17120060 TI - Cardiovascular complications of pediatric chronic kidney disease. AB - Cardiovascular disease (CVD) mortality is a leading cause of death in adult chronic kidney disease (CKD), with exceptionally high rates in young adults, according to the Task Force on Cardiovascular Disease. Recent data indicate that cardiovascular complications are already present in children with CKD. This review summarizes the current literature on cardiac risk factors, mortality and morbidity in children with CKD. PMID- 17120062 TI - The association of anemia and hypoalbuminemia with accelerated decline in GFR among adolescents with chronic kidney disease. AB - We sought to describe rates of kidney function decline and to identify modifiable risk factors for CKD progression in a multicenter prospective cohort study of adolescents with CKD aged 11 to 18 years seen semiannually for up to three years. Of the 23 subjects meeting inclusion criteria, the average estimated GFR was 51 +/- 27 ml/min/1.73 m(2) (0.85 +/- 0.45 ml/s/1.73 m(2)) at entry. The overall annualized decline in GFR was 5.6 ml/min/1.73 m(2) (0.093 ml/s/1.73 m(2)) per year (95% confidence interval [95% CI]: 1.9 to 9.3 [0.032 to 0.16]). The adjusted annualized decline in GFR was found to be accelerated in males, as well as among those over 15 years of age. The adjusted annualized decline in GFR was greater among those with either anemia (hematocrit below 36%), or hypoalbuminemia (albumin below 4 g/dl [40 g/L]). After adjustment, anemia was associated with an accelerated decline of 7.8 ml/min/1.73 m(2) (0.13 ml/s/1.73 m(2)) (95% CI: 3.3 to 12 [0.055 to 0.20]) and hypoalbuminemia was associated with an accelerated decline of 17 ml/min/1.73 m(2) (0.28 ml/s/1.73 m(2)) (95% CI: 11 to 22 [0.18 to 0.37]). Further study is needed to evaluate whether treatment of anemia or hypoalbuminemia, as outlined in current clinical care guidelines for CKD, may slow the progression of CKD in adolescents. PMID- 17120063 TI - Examining Ambrosia pollen episodes at Poznan (Poland) using back-trajectory analysis. AB - The pollen grains of Ambrosia spp. are considered to be important aeroallergens in parts of southern and central Europe. Back-trajectories have been analysed with the aim of finding the likely sources of Ambrosia pollen grains that arrived at Poznan (Poland). Temporal variations in Ambrosia pollen at Poznan from 1995 2005 were examined in order to identify Ambrosia pollen episodes suitable for further investigation using back-trajectory analysis. The trajectories were calculated using the transport model within the Lagrangian air pollution model, ACDEP (Atmospheric Chemistry and Deposition). Analysis identified two separate populations in Ambrosia pollen episodes, those that peaked in the early morning between 4 a.m. and 8 a.m., and those that peaked in the afternoon between 2 p.m. and 6 p.m.. Six Ambrosia pollen episodes between 2001 and 2005 were examined using back-trajectory analysis. The results showed that Ambrosia pollen episodes that peaked in the early morning usually arrived at Poznan from a southerly direction after passing over southern Poland, the Czech Republic, Slovakia and Hungary, whereas air masses that brought Ambrosia pollen to Poznan during the afternoon arrived from a more easterly direction and predominantly stayed within the borders of Poland. Back-trajectory analysis has shown that there is a possibility that long-range transport brings Ambrosia pollen to Poznan from southern Poland, the Czech Republic, Slovakia and Hungary. There is also a likelihood that Ambrosia is present in Poland, as shown by the arrival of pollen during the afternoon that originated primarily from within the country. PMID- 17120064 TI - Risk assessment of the gypsy moth, Lymantria dispar (L), in New Zealand based on phenology modelling. AB - The gypsy moth is a global pest that has not yet established in New Zealand despite individual moths having been discovered near ports. A climate-driven phenology model previously used in North America was applied to New Zealand. Weather and elevation data were used as inputs to predict where sustainable populations could potentially exist and predict the timing of hatch and oviposition in different regions. Results for New Zealand were compared with those in the Canadian Maritimes (New Brunswick, Nova Scotia, and Prince Edward Island) where the gypsy moth has long been established. Model results agree with the current distribution of the gypsy moth in the Canadian Maritimes and predict that the majority of New Zealand's North Island and the northern coastal regions of the South Island have a suitable climate to allow stable seasonality of the gypsy moth. New Zealand's climate appears more forgiving than that of the Canadian Maritimes, as the model predicts a wider range of oviposition dates leading to stable seasonality. Furthermore, we investigated the effect of climate change on the predicted potential distribution for New Zealand. Climate change scenarios show an increase in probability of establishment throughout New Zealand, most noticeably in the South Island. PMID- 17120065 TI - Statistical modeling of valley fever data in Kern County, California. AB - Coccidioidomycosis (valley fever) is a fungal infection found in the southwestern US, northern Mexico, and some places in Central and South America. The fungus that causes it (Coccidioides immitis) is normally soil-dwelling but, if disturbed, becomes air-borne and infects the host when its spores are inhaled. It is thus natural to surmise that weather conditions that foster the growth and dispersal of the fungus must have an effect on the number of cases in the endemic areas. We present here an attempt at the modeling of valley fever incidence in Kern County, California, by the implementation of a generalized auto regressive moving average (GARMA) model. We show that the number of valley fever cases can be predicted mainly by considering only the previous history of incidence rates in the county. The inclusion of weather-related time sequences improves the model only to a relatively minor extent. This suggests that fluctuations of incidence rates (about a seasonally varying background value) are related to biological and/or anthropogenic reasons, and not so much to weather anomalies. PMID- 17120066 TI - Health-related quality of life and predicting survival in cancer: not a simple matter. PMID- 17120067 TI - The relationship between dyspnea and patient satisfaction with quality of life in advanced cancer. AB - GOALS OF THE WORK: Dyspnea is a common symptom in patients with advanced cancer. The goal of this study was to investigate the relationship between dyspnea and patient satisfaction with quality of life (QoL) in advanced cancer. MATERIALS AND METHODS: A case series of 954 cancer patients treated at the Cancer Treatment Centers of America was investigated. Dyspnea was measured using the EORTC dyspnea subscale. Patient satisfaction with QoL was measured using the Ferrans and Powers Quality of Life Index (QLI). The relationship between dyspnea and QLI was evaluated using multivariate analysis of variance and multiple regression analysis. RESULTS: Of 954 patients, 579 were females and 375 males with the median age at presentation of 56 years (range: 20-90 years). Of these patients, 66% did not respond to prior treatment. Most common cancers were breast (26%), colorectal (19%) and lung (16%). After controlling for the effects of age and treatment history, every ten unit increase in dyspnea was statistically significantly associated with 0.81, 0.16, 0.47, and 0.47 unit decline in QLI health/physical, social/economic, psychological/spiritual and global function score, respectively. CONCLUSIONS: We found that dyspnea is strongly correlated with patient satisfaction with QoL in advanced cancer. Future studies should evaluate the impact of integrative cancer care services on patient satisfaction with QoL. PMID- 17120068 TI - Breast cancer survivors' supportive care needs 2-10 years after diagnosis. AB - GOALS OF THE WORK: A significant proportion of breast cancer patients experience psychosocial morbidity after treatment, although their longer-term outcomes and supportive care service needs have not been comprehensively documented. The aim of this study was to identify longer-term outcomes and supportive care needs in disease-free breast cancer survivors. MATERIALS AND METHODS: One hundred seventeen patients who had been diagnosed with breast cancer 2-10 years earlier completed questionnaires to assess psychosocial outcomes including supportive care needs, psychological distress, and quality of life (QoL). MAIN RESULTS: QoL and depression scores were consistent with community rates although anxiety scores were higher. Approximately two thirds of survivors reported at least one unmet need, most frequently concerning existential survivorship issues, thereby highlighting the unique needs of survivors. Years since diagnosis was not correlated with need levels. Survivors classified as clinically anxious reported over three times as many unmet needs and survivors classified as depressed reported over two and a half times as many unmet needs. Positive outcomes were frequently reported. CONCLUSIONS: The findings have direct clinical relevance: irrespective of years since diagnosis, comprehensive and extended supportive care services are required to identify breast cancer survivors in need of supportive care interventions and remediate high levels of anxiety. PMID- 17120069 TI - The development and validation of a prediction tool for chemotherapy-induced anemia in patients with advanced nonsmall cell lung cancer receiving palliative chemotherapy. AB - BACKGROUND: In this study, the development and validation of a cycle-based prediction model for severe anemia [i.e., a hemoglobin (Hb) of or=8 to <10 was identified as the optimal cut off to maximize the sensitivity (83.1%) and specificity (67.8%) of the prediction tool. Patients with a score of >or=8 would be considered at high risk for developing anemia after a particular cycle of chemotherapy. DISCUSSION: We developed and validated an anemia prediction tool for advanced stage NSCLC patients receiving palliative chemotherapy. To make the model available for easy use and access, we have incorporated it on to our risk prediction website: http://www.PredictPatientEvents.com . It is hoped that this risk model will enhance patient care by optimizing the frequency of Hb testing and/or the use of preventative therapies. PMID- 17120070 TI - How important is the opinion of significant others to cancer patients' adjuvant chemotherapy decision-making? AB - GOALS: Decisions regarding adjuvant chemotherapy are difficult, since value tradeoffs are involved. Little is known about the importance of the significant others in patients' decision-making regarding adjuvant treatment. We surveyed patients with breast and colorectal cancer about the importance they assigned to the opinions of their significant others and assessed correlates of these importance scores. MATERIALS AND METHODS: One hundred and twenty-three patients rated on a five-point scale how much they cared about the opinion of six significant others. MAIN RESULTS: Most important was the opinion of their treating specialist, followed by that of their partner, children, other family, friends, and colleagues. Women assigned higher scores to the opinion of their children, younger patients to that of their specialist, and patients who were about to undergo chemotherapy to that of their family. Patients with breast cancer and patients without paid employment assigned slightly more importance to the opinion of their partner. CONCLUSIONS: Information on the influence of significant others may help clinicians when involving patients in treatment decision-making and discussing patients' treatment preferences. PMID- 17120071 TI - Prospective study of standalone balloon kyphoplasty with calcium phosphate cement augmentation in traumatic fractures. AB - Prospective consecutive series cases study to investigate the clinical and radiological results of standalone balloon kyphoplasty and cement augmentation with calcium phosphate in traumatic fractures. Independent observer evaluation of radiological and computer tomography results, visual analogue scale (VAS), Roland Morris score and complications with acute traumatic compression fractures type A, treated with a standalone balloon kyphoplasty and cement augmentation with calcium phosphate (Calcibon); follow-up time at a mean of 30 months (24-37 months). From August 2002 to August 2003, consecutive patients with traumatic compression fractures (Magerl type A) without neurological deficit underwent standalone kyphoplasty with Calcibon. We report here the pre-, post-operative and the follow-up results, applying the VAS (0-10) for pain rating, the Roland-Morris (0-24) disability score, CT-scan examination, detailed radiographic evaluation of vertebral body (VB) deformity and segmental kyphosis measurement. The pre operative X-ray measurements, VAS and the 7 days Roland-Morris scores are compared with the post-operative and the 30 months follow-up findings. Twenty eight patients with 33 treated fracture levels were included in this study. The mean initial vertebral deformity (VB kyphosis) was 17 degrees, corrected to a post-operative of 6 degrees. We noted a loss of correction at the follow-up in comparison to the post-operative standing X-ray at 24 h of 3 degrees vertebral deformity and 3 degrees segmental kyphosis. The VAS score demonstrates a decrease over time from a mean of 8.7-3.1 at 7 days and to 0.8 at the last follow-up. The Roland-Morris disability score demonstrates a similar improvement. We noticed no major complications related to the procedure. The mean cement resorption after 1 year was 20.3% (0.3-35.3%) and is related to the individual biological resorption process and is not predictable. All patients with vertebral fractures as sole medical problem were discharged within 48 h. All active patients returned to the same work within 3 months with the same working ability as before the accident. Standalone balloon kyphoplasty is a potential alternative mini-invasive technique to reduce the fractures. However, due to the intrinsic characteristic of calcium phosphate cement (Calcibon) we recommend the application of this biological cement for standalone reduction and stabilisation only in fractures type A1 and A3.1 in young patient. In case of higher destruction levels of the VB, we propose the utilisation of Calcibon associated with posterior instrumentation. Having regard to the pointed out indications, our preliminary results demonstrate a new possibility to treat this kind of fractures, allowing a rapid handling of pain, early discharge and return to normal activities. PMID- 17120072 TI - Lumbar spondylolysis: a life long dynamic condition? A cross sectional survey of 4.151 adults. AB - Lumbar spondylolysis (LS) has been the subject of several studies focusing on adolescent athletes. Few, if any, studies have examined LS in the general population. Lysis of the pars interarticularis of the vertebra may be associated with slipping (olisthesis), or it may be stable. In the present survey of lumbar radiographs and general epidemiological data recorded from the Copenhagen Osteoarthritis Study cohort of 4.151 subjects (age range, 22-93 years), we identified the distribution and individual risk factors for LS-development. Men were significantly more at risk of L5 spondylolysis (P = 0.002). There were no sex-specific significant differences regarding LS-incidence at the L4 level. We found no significant differences of risk of LS between nulliparous or multiparous women (L4 P = 0.54/L5 P = 0.35). Furthermore, we found no significant relationship between age at menopause and LS-development. Increased lumbar lordosis was associated to L4/L5 spondylolysis in men (L4 P < 0.001/L5 P = 0.008). In women increased lumbar lordosis had a significant association with L5 spondylolysis (P < 0.001). Increased pelvic inclination was associated with L5 spondylolysis in both men and women (P < 0.001). There were no sex-specific differences regarding the occurrence of simultaneous slips/non-slips. In men, no individual risk factors for L4 slips with concomitant LS were found. In women slipped LS of L4 were significantly associated to aging (P < 0.001) and with decreased pelvic inclination (P = 0.001). In men slipped LS of L5 was significantly associated to increased BMI (P = 0.002), but not to aging (P = 0.10). In women, slips of L5 LS were significantly correlated to aging (P = 0.005), to BMI recorded at the time of radiographic examination (P = 0.006), and BMI measured 17 years before radiographic index examination (P = 0.004). The present study contrasts with commonly held views regarding lumbar spondylolysis. The prevalence of LS increases throughout life and is apparently not a condition restricted to adolescence. Although the cross-sectional nature of the present study prevents an exact estimate of the age at onset; future, sequential studies of the cohort may provide us with some important answers on this topic. Apart from aging-obesity, lordotic angle and pelvic inclination were found to be individual risk factors for LS. PMID- 17120073 TI - Conformational stability and dynamics of cytochrome c affect its alkaline isomerization. AB - The alkaline isomerization of horse heart ferricytochrome c (cyt c) has been studied by electronic absorption spectroscopy in the presence of the Hofmeister series of anions: chloride, bromide, rhodanide and perchlorate. The anions significantly affect the apparent pK (a) value of the transition in a concentration-dependent manner according to their position in the Hofmeister series. The Soret region of the absorption spectra is not affected by the presence of the salts and shows no significant structural perturbation of the heme crevice. In the presence of perchlorate and rhodanide anions, the cyanide exchange rate between the bulk solvent and the binding site is increased. These results imply higher flexibility of the protein structure in the presence of chaotropic salts. The thermal and isothermal denaturations monitored by differential scanning calorimetry and circular dichroism, respectively, showed a decrease in the conformational stability of cyt c in the presence of the chaotropic salts. A positive correlation between the stability, DeltaG, of cyt c and the apparent pK (a) values that characterize the alkaline transition indicates the presence of a thermodynamic linkage between these conformational transitions. In addition, the rate constant of the cyanide binding and the partial molar entropies of anions negatively correlate with the pK (a) values. This indicates the important role of anion-induced solvent reorganization on the structural flexibility of cyt c in the alkaline transitions. PMID- 17120074 TI - Quantification of single-stranded nucleic acid and oligonucleotide interactions with metal ions by affinity capillary electrophoresis: part I. AB - The interactions between oligonucleotides and inorganic cations have been measured by capillary zone electrophoresis. With increasing concentrations of divalent cations (Ca(2+), Mg(2+), Mn(2+) and Ni(2+)) in the running buffer, the migration behavior was evaluated by calculation of the binding constants. Besides these fundamental studies of binding equilibria, different buffer components, tris(hydroxymethyl)aminomethane and 3-(N-morpholino)propanesulfonic acid, have been investigated and their effects on metal ion binding quantified. PMID- 17120075 TI - Investigations of differences in iron oxidation state inside single neurons from substantia nigra of Parkinson's disease and control patients using the micro XANES technique. AB - X-ray absorption near edge structure spectroscopy was applied in order to investigate differences in iron chemical state between the nerve cells of substantia nigra (SN) representing Parkinson's disease (PD) and those of control cases. Autopsy samples were cut using a cryotome, and were not fixed and not embedded in paraffin. The comparison of the absorption spectra near the iron K edge measured in melanized neurons from SN of PD and control samples did not show significant differences in iron oxidation state. Measurements of inorganic reference materials containing iron in the second and third oxidation states indicate that most of the iron in all the nerve cell bodies examined was oxidized and occurred as trivalent ferric iron (Fe(3+)). PMID- 17120077 TI - Interfacial and surface characterization of two self-etching adhesive systems and a total-etch adhesive after bonding to ground and unground bovine enamel--a qualitative study. AB - The purpose of the study was to evaluate the enamel surface and interface morphology of two self-etching adhesive systems (SAS) vs a total-etch control, after bonding to ground and unground enamel using field emission scanning electron microscopy (FESEM). Thirty bovine incisors were used in this study. The buccal enamel surface of 15 teeth was ground flat to resemble freshly cut enamel. The rest of the teeth were left intact. Two SAS, Clearfil SE Bond (CSE, Kuraray) and Prompt L-Pop (3M-ESPE), and a conventional adhesive system, Scotchbond Multipurpose (3M-ESPE, control), were used to condition the surface of unground and ground enamel on 12 teeth. A composite button was bonded to the remaining 18 teeth; a cross-section (1 mm thick) was obtained from each and the bonded interface was polished. All specimens were dehydrated in ascending grades of ethanol, gold-sputter-coated, and observed under FESEM (Hitachi S-4000) to evaluate the ultrastructural morphology of the enamel surface and the enamel dentin interface. The etching patterns and adhesive penetration varied according to the aggressiveness of the SAS, with CSE being the mildest and H3PO4 being the most aggressive. There were no significant differences on the ultrastructural morphology of the enamel surface between unground and ground specimens. It appears that microporosities within enamel prisms provide sufficient enamel-resin hybridization in unground enamel. The enamel dissolution pattern and depth of infiltration depend on the type of SAS used, with no significant differences in unground and ground enamel. PMID- 17120078 TI - Ligand specificity of odorant receptors. AB - Odorant receptors belong to class A of the G protein-coupled receptors (GPCRs) and detect a large number of structurally diverse odorant molecules. A recent structural bioinformatic analysis suggests that structural features are conserved across class A of GPCRs in spite of their low sequence identity. Based on this work, we have aligned the sequences of 29 ORs for which ligand binding data are available. Recent site-directed mutagenesis experiments on one such receptor (MOR174-9) provide information that helped to identify nine amino-acid residues involved in ligand binding. Our modeling provides a rationale for amino acids in equivalent positions in most of the odorant receptors considered and helps to identify other amino acids that could be important for ligand binding. Our findings are consistent with most of the previous models and allow predictions for site-directed mutagenesis experiments, which could also validate our model. PMID- 17120079 TI - Effects of a metacognitive intervention on students' approaches to learning and self-efficacy in a first year medical course. AB - AIM: To determine the influence of metacognitive activities within the PBL tutorial environment on the development of deep learning approach, reduction in surface approach, and enhancement of individual learning self-efficacy. METHOD: Participants were first-year medical students (N = 213). A pre-test, post-test design was implemented with intervention and control cohorts, with intervention students experiencing a program of metacognitive activities within their PBL tutorials of at least 20 weeks duration. All students completed the Medical Course Learning Questionnaire at the commencement, and again at the completion of, the study. The metacognitive intervention itself consisted of reflection on the learning in PBL coupled with peer- and self-assessment. RESULTS: Self efficacy was significantly reduced for both control and intervention cohorts at the conclusion of the study. A significant reduction in the adoption of deep and strategic learning approach, matched by a corresponding increase in the use of surface learning, was demonstrated for both cohorts. There was a statistically significant association between high self-efficacy and deep learning approach, with older students over-represented in the group of efficacious deep learners. CONCLUSION: Over the course of first-year medical studies, students lose self efficacy and move away from deep-strategic learning approaches towards more surface approaches. The program of metacognitive activities failed to reverse this trend. The substantial swing towards surface learning raises questions about the perceived capacity of PBL curricula to promote deep approaches to learning in dense curricula, and reinforces the importance of personal and contextual factors, such as study habits, workload and assessment, in determining individual approaches and idiosyncratic responses to learning situations. PMID- 17120080 TI - How clinical instructors can enhance the learning experience of physical therapy students in an introductory clinical placement. AB - PURPOSE: There is little understanding of how physical therapy students are influenced by clinical instructors (CIs) particularly at the outset of their clinical learning. The purpose of this study was to evaluate physical therapy students' perceptions of their learning experiences during an introductory clinical placement. METHODS: Subjects were 51 physical therapy students participating in an introductory clinical placement following the first unit (semester) of a 2-year, entry-level Master's program. Each day of their placement, students completed a questionnaire based on Brookfield's Critical Incident Questionnaire. Students were asked when they felt most engaged or distanced, what actions of others were most affirming or puzzling, and what surprised them about the placement that day. Two evaluators independently read and coded the entries in all questionnaires, and then collaborated to establish themes related to the behaviors of the CI. RESULTS: The CI could enrich the students' experiences in the following ways: (1) prepare them by introducing, explaining, demonstrating, or allowing them time to obtain information. (2) Confirm learning by providing feedback and recapping. (3) Provide "hands on" experiences appropriate to students' knowledge, skills and comfort. (4) Challenge students by questioning, discussing possibilities, or providing time for reflection. (5) Respect students, value their input and allow them an appropriate level of independence. (6) Demonstrate professional behavior related to communication, evidence-based practice and continuing education. CONCLUSIONS: Physical therapy students value CIs who involve them in patient care; confirm, challenge and prepare them for learning; respect their input and model professional behavior. PMID- 17120081 TI - Epithelial cells treated with genistein inhibit adhesion and endocytosis of Paracoccidioides brasiliensis. AB - Paracoccidioidomycosis is caused by Paracoccidioides brasiliensis, which although not formally considered an intracellular pathogen, can be internalized by epithelial cells in vitro and in vivo. The mechanisms used by P. brasiliensis to adhere to and invade non-professional phagocytes have not been identified. The signal-transduction networks, involving protein tyrosine kinase (PTK) and protein phosphatase activities, can modulate crucial events during fungal infections. In this study, the involvement of PTK has been investigated in P. brasiliensis adherence and invasion in mammalian epithelial cells. A significant inhibition of the fungal invasion occurred after the pre-treatment of the epithelial cells with genistein, a specific tyrosine kinase inhibitor, indicating that the tyrosine kinase pathway is involved in P. brasiliensis internalization. In contrast, when the fungus was treated, a slight (not significant) inhibition of PTK was observed, suggesting that PTK might not be the fungus' transduction signal pathway during the invasion process of epithelial cells. An intense PTK immunofluorescence labeling was observed in the periphery of the P. brasiliensis infected cells, little PTK labeling was found in both uninfected cells and yeast cells, at later infection times (8 and 24 h). Moreover, when the epithelial cells were treated with genistein and infected with P. brasiliensis, no labeling was observed, suggesting the importance of the PTK in the infectious process. These results suggest that PTK pathway participates in the transduction signal during the initial events of the adhesion and invasion processes of P. brasiliensis to mammalian epithelial cells. PMID- 17120082 TI - Effect of Agave tequilana juice on cell wall polysaccharides of three Saccharomyces cerevisiae strains from different origins. AB - In this study, a characterization of cell wall polysaccharide composition of three yeasts involved in the production of agave distilled beverages was performed. The three yeast strains were isolated from different media (tequila, mezcal and bakery) and were evaluated for the beta(1,3)-glucanase lytic activity and the beta-glucan/ mannan ratio during the fermentation of Agave tequilana juice and in YPD media (control). Fermentations were performed in shake flasks with 30 g l(-1) sugar concentration of A. tequilana juice and with the control YPD using 30 g l(-1) of glucose. The three yeasts strains showed different levels of beta-glucan and mannan when they were grown in A. tequilana juice in comparison to the YPD media. The maximum rate of cell wall lyses was 50% lower in fermentations with A. tequilana juice for yeasts isolated from tequila and mezcal than compared to the bakery yeast. PMID- 17120083 TI - Influence of prey on developmental performance, reproduction and prey consumption of Neoseiulus californicus (Acari: Phytoseiidae). AB - The Spical strain of the predatory mite Neoseiulus californicus (McGregor) is used as a biological control agent, but little is known about its preferred prey and host plants in Japan. Here we studied the development, reproduction and prey consumption of the Spical strain when fed on eggs of five different spider mite species deposited on both their laboratory-rearing plant and cherry, on which all five spider mite species developed well. The developmental periods of immature N. californicus females and males were significantly affected by the prey species they fed on, but not by the plants. No difference was found between males and females. The developmental period was shorter on eggs of two Tetranychus species than on eggs of Panonychus ulmi. Immature females had a higher predation rate than immature males. Preoviposition period, oviposition period and the number of eggs laid per female were not significantly affected by either the plants or the type of prey eggs. The postoviposition period and total adult longevity were shorter on eggs of P. ulmi than of the other four prey species, but there was no effect of plant substrate. The postoviposition period of the Spical strain was much longer than that of other N. californicus strains or other predatory mite species: the postoviposition period of the Spical strain was more than three times longer than the oviposition period, accounting for more than 75% of the total adult longevity. This suggests that the females need multiple mating to reach full egg load, but this remains to be tested. Total consumption by N. californicus adults was lower for eggs of P. ulmi than for eggs of the other four species, apparently because of the shorter postoviposition period when fed on eggs of P. ulmi. The intrinsic rates of natural increase (r(m)) on the rearing plant did not differ among prey species, whereas those on cherry were significantly different: the value was higher on Tetranychus urticae eggs than on eggs of other species. Only when N. californicus fed on T. urticae eggs, the r(m) values were significantly different between the rearing plant and cherry (higher on cherry). Thus, the Spical strain of N. californicus could feed on eggs of all five spider mite species, deposited on a variety of plants with similar r(m) values, suggesting that it could be successfully used to control spider mites in orchards and various crop fields of Japan. PMID- 17120084 TI - PTH regulation of c-Jun terminal kinase and p38 MAPK cascades in intestinal cells from young and aged rats. AB - In the present study, we examined the role of Parathyroid hormone (PTH) on the c Jun N-terminal kinase (JNK) 1/2 and p38 mitogen-activated protein kinase (MAPK) members of the MAPK family as it relates to ageing by measuring hormone-induced changes in their activity in enterocytes isolated from young (3 month old) and aged (24 month old) rats. Our results show that PTH induces a transient activation of JNK 1/2, peaking at 1 min (+threefold). The hormone also stimulates JNK 1/2 tyrosine phosphorylation, in a dose-dependent fashion, this effect being maximal at 10 nM. PTH-induced JNK 1/2 phosphorylation was suppressed by its selective inhibitor SP600125. Moreover, hormone-dependent activation of JNK 1/2 was dependent on calcium, since pretreatment of cells with BAPTA-AM or EGTA blocked PTH effects. With ageing, the response to PTH was significantly reduced. JNK basal protein expression was not different in the enterocytes from young and aged rats, however, basal protein phosphorylation increased with ageing. PTH did not stimulate, within 1-10 min, the basal activity and phosphorylation of p38 MAPK in rat intestinal cells. The hormone increased enterocyte DNA synthesis; the response was dose-dependent and decreased (-40%) with ageing. In agreement with the mitogenic role of the MAPK cascades, this effect was blocked by specific inhibitors of extracellular signal-regulated protein kinase (ERK) 1/2 and JNK 1/2. The results obtained in this work expand our knowledge on the mechanism of action of PTH in duodenal cells. PMID- 17120085 TI - Chondrogenesis of human periosteum-derived progenitor cells in atelocollagen. AB - Periosteum-derived progenitor cells (PDPCs) could be differentiated into cartilage using atelocollagen as a carrier and in the presence of transforming growth factor-beta3 (TGF-beta3). Chondrogenesis was verified by RT-PCR and Western blotting. Expression of the type II collagen mRNA was found from the differentiated PDPCs in atelocollagen 3 weeks after chondrogenic induction. The chondrogenic potential of the PDPCs was also verified by histochemical staining for type II collagen protein. Increased production of glycosaminoglycan shows that the PDPCs in atelocollagen could differentiate into chondrocytes under a chondrogenic environment. PDPCs can therefore be used as a cell source for cell based therapies targeted toward the articular cartilage of the knee. PMID- 17120086 TI - pEGFP-T, a novel T-vector for the direct, unidirectional cloning and analysis of PCR-amplified promoters. AB - A novel T-vector, designated as pEGFP-T, was constructed which could be used for direct and unidirectional cloning and analysis of promoters. The method involves minimizing the non-recombinant background of the T-vector when used to clone PCR products, and maintaining only the forward orientation of the PCR products prior to transformation. The usefulness of this vector is demonstrated in cloning and analyzing promoters. PMID- 17120087 TI - Antibody production with yeasts and filamentous fungi: on the road to large scale? AB - Yeasts and filamentous fungi have gained significant interest for the production of recombinant antibodies and antibody fragments. The opportunities and constraints of antibody (fragment) production in these hosts are highlighted as well as cell engineering strategies to overcome the constraints. Following aspects are addressed: folding, assembly and secretion of antibody related proteins, process optimization to improve productivity and quality, proteolysis, and, as a major point of interest, glycosylation. PMID- 17120088 TI - Transgenic wine yeast technology comes of age: is it time for transgenic wine? AB - Saccharomyces cerevisiae is the main yeast responsible for alcoholic fermentation of grape juice during wine making. This makes wine strains of this species perfect targets for the improvement of wine technology and quality. Progress in winemaking has been achieved through the use of selected yeast strains, as well as genetic improvement of wine yeast strains through the sexual and pararexual cycles, random mutagenesis and genetic engineering. Development of genetically engineered wine yeasts, their potential application, and factors affecting their commercial viability will be discussed in this review. PMID- 17120089 TI - Changes in primary metabolism leading to citric acid overflow in Aspergillus niger. AB - For citric acid-accumulating Aspergillus niger cells, the enhancement of anaplerotic reactions replenishing tricarboxylic acid cycle intermediates predisposes the cells to form the product. However, there is no increased citrate level in germinating spores and a complex sequence of developmental events is needed to change the metabolism in a way that leads to an increased level of tricarboxylic acid cycle intermediates in mycelia. A review of physiological events that cause such intracellular conditions, with the special emphasis on the discussion of hexose transport into the cells and regulation of primary metabolism, predominantly of glycolytic flux during the process, is presented. PMID- 17120090 TI - Use of molecular markers and flow cytometry to preserve ancient Annurca apple germplasm. AB - The old Annurca apple cultivar (Malus domestica), particularly appreciated for its peculiar flavor and crispy flesh, was studied in order to preserve its ancient germplasm. Twelve clones of Annurca were analyzed using random amplified polymorphic DNA (RAPD) and simple-sequence repeat (SSR) markers. Two out of 30 RAPD primers and nine out of ten SSR primers were able to discriminate all the clones analyzed. Data were confirmed by measuring DNA content using flow cytometry. The results provide a good procedure to improve germplasm field management, in order to removing redundant material in the Annurca collection. This represents an efficient way to create a data bank in order to preserve the genetic variability of the Annurca cultivar. PMID- 17120091 TI - Preparation of validoxylamine A by biotransformation of validamycin A using resting cells of a recombinant Escherichia coli. AB - A gene encoding beta-glucosidase was cloned and over-expressed in Escherichia coli. Validamycin A was then biotransformed into validoxylamine A by using the resting recombinant cells. The biotransformation yield reached 92% when the reaction was performed at 37 degrees C for 1 h in the presence of 100 ml sodium phosphate buffer (0.1 M, pH 7.0), 32 mM validamycin A and 0.71 mg dry cell w/ml. PMID- 17120092 TI - Ca2+ and Cu2+ supplementation increases mannitol production by Candida magnoliae. AB - Supplementation with CaCl(2).2H(2)O (50 mg l(-1)) or CuSO(4).5H(2)O (10 mg l(-1)) improved mannitol production by Candida magnoliae by 14.5 and 18.6% (25 and 32 g/L), respectively. When used in combination, they acted synergistically: Ca(2+) decreased the intracellular concentration of mannitol 30%, whereas Cu(2+ )increased the intracellular activity of mannitol dehydrogenase 1.6-times more than control. Ca(2+) probably works by altering the permeability of cells to mannitol, whereas, Cu(2+) increases the activity of an enzyme responsible for mannitol biosynthesis. PMID- 17120093 TI - Streptomycin production by Streptomyces griseus can be modulated by a mechanism not associated with change in the adpA component of the A-factor cascade. AB - In Streptomyces coelicolor, AtrA is an activator of transcription of the actinorhodin cluster-situated regulator gene actII-ORF4. In previous work, we showed that S. coelicolor AtrA binds in vitro to the promoter of S. griseus strR, the streptomycin cluster-situated regulator. We show here that S. griseus carries a single close homologue of atrA and that expression of S. coelicolor AtrA in S. griseus causes a DNA binding-dependent reduction in streptomycin production and in the mRNA levels of strR and genes of streptomycin biosynthesis. However, there is no effect on the level of the mRNA of adpA, which is the only transcription factor that has so far been characterised for strR. The adpA gene is directly regulated by ArpA, the receptor protein for the gamma-butyrolactone signalling molecule A-factor. Therefore, to our knowledge, our results provide the first in vivo evidence that A-factor-ArpA-AdpA-StrR regulatory cascade represents only part of the full complexity of regulation of streptomycin biosynthesis in S. griseus. The potential biotechnological application of our findings is discussed. PMID- 17120094 TI - Fluorophenol oxidation by a fungal chloroperoxidase. AB - Caldariomyces fumago chloroperoxidase degrades monofluorophenols at both pH 3 and pH 6. 4-Fluorophenol is most readily degraded and its oxidation is most efficient at pH 6. GC-MS analyses of the reaction products revealed compounds relating to the reaction of fluorophenol radical. The degradation of fluorinated compounds is of significant environmental interest and this versatile enzyme may by employed to treat contaminated soil or water prior to discharge. PMID- 17120095 TI - Oxygen transfer model in recombinant Pichia pastoris and its application in biomass estimation. AB - An oxygen transfer model was established for Pichia pastoris growing on glycerol and methanol in a stirred tank bioreactor and expressing a recombinant human serum albumin (rHSA). This was based on pseudo-steady state mass balance, where the volumetric O(2) transfer coefficient, k (L) a, was estimated as a function of power input per unit volume and aeration rate. Under pseudo-steady state, the O(2) transfer rate model matched the O(2) uptake rate obtained from a previous macrokinetic model. This procedure was also applied to estimate biomass concentration by using the on-line rolling identification approach. PMID- 17120096 TI - Degradation of 2,5-dimethylpyrazine by Rhodococcus erythropolis strain DP-45 isolated from a waste gas treatment plant of a fishmeal processing company. AB - A bacterium, strain DP-45, capable of degrading 2,5-dimethylpyrazine (2,5-DMP) was isolated and identified as Rhodococcus erythropolis. The strain also grew on many other pyrazines found in the waste gases of food industries, like 2,3 dimethylpyrazine (2,3-DMP), 2,6-dimethylpyrazine (2,6-DMP), 2-ethyl-5(6) dimethylpyrazine (EMP), 2-ethylpyrazine (EP), 2-methylpyrazine (MP), and 2,3,5 trimethylpyrazine (TMP). The strain utilized 2,5-DMP as sole source of carbon and nitrogen and grew optimally at 25 degrees C with a doubling time of 7.6 h. The degradation of 2,5-DMP was accompanied by the growth of the strain and by the accumulation of a first intermediate, identified as 2-hydroxy-3,6 dimethylpyrazine (HDMP). The disappearance of HDMP was accompanied by the release of ammonium into the medium. No other metabolite was detected. The degradation of 2,5-DMP and HDMP by strain DP-45 required molecular oxygen. The expression of the first enzyme in the pathway was induced by 2,5-DMP and HDMP whereas the second enzyme was constitutively expressed. The activity of the first enzyme was inhibited by diphenyliodonium (DPI), a flavoprotein inhibitor, methimazole, a competitive inhibitor of flavin-containing monooxygenases, and by cytochrome P450 inhibitors, 1-aminobenzotriazole (ABT) and phenylhydrazine (PHZ). The activity of the second enzyme was inhibited by DPI, ABT, and PHZ. Sodium tungstate, a specific antagonist of molybdate, had no influence on growth and consumption of 2,5-DMP by strain DP-45. These results led us to propose that a flavin-dependent monooxygenase or a cytochrome P450-dependent monooxygenase rather than a molybdenum hydroxylase catalyzed the initial hydroxylation step and that a cytochrome P450 enzyme is responsible for the transformation of HDMP in the second step. PMID- 17120097 TI - Isolation of the subclavian artery: 4 cases report and literature review. PMID- 17120098 TI - Prevalence of concomitant atherosclerotic arterial diseases in patients with significant cervical carotid artery stenosis in Taiwan. AB - BACKGROUND: Atherosclerotic stenosis of extracranial carotid arteries is an important cause of ischemic stroke in Taiwan, an ethnic Chinese population. Concurrent atherosclerotic arterial disease is an important issue in the management of patients with carotid stenosis, but its prevalence and extent are unknown. METHODS: One hundred and sixty-three consecutive patients with angiographically proven significant cervical carotid artery stenosis (>or=50% stenosis) were enrolled in this study. Angiography was done to document concurrent coronary, renal, and limb artery stenosis. Clinical symptoms and signs were also correlated with the angiographic findings. RESULTS: One or more significant concurrent arterial stenotic disease was found in 73% of the patients. The most frequent were coronary artery disease, found in 68% of the patients, while renal artery stenosis and limb artery stenosis were found in 20% and 21% of the patients, respectively. Age, diabetes, history of angina pectoris, intermittent claudication, and asymmetric arm blood pressures were significantly associated with the presence of concurrent arterial stenosis. However, 41% of the patients with concurrent coronary artery disease did not have any clinical symptoms or history of myocardial infarction. CONCLUSIONS: Our data indicated that concurrent advanced and extensive arterial disease is common in patients with significant cervical carotid stenosis, and also suggest the importance of global evaluation of systemic atherosclerosis in these patients to achieve optimal management. PMID- 17120100 TI - Major and trace elements in paddy soil contaminated by Pb-Zn mining: a case study of Kocani Field, Macedonia. AB - The objective of this study was to assess the bulk chemical composition as well as the extent and severity of heavy metal contamination in the paddy soil of Kocani Field (eastern Macedonia). The results revealed that the paddy soil of the western part of Kocani Field is severely contaminated with Pb, Zn, As and Cd in the vicinity of the Zletovska River due to irrigation with riverine water that is severely affected by acid mine and tailing effluents from the Pb-Zn mine in Zletovo. The detected total concentrations of these metals are far above the threshold values considered to be phytotoxically excessive for surface soil. The paddy soil in the vicinity of the Zletovska River was also found to exhibit elevated levels of Ba, Th, U, V, W, Mo, Cu, Sb, Bi, Ag, Au, Hg and Tl, with concentrations above their generally accepted median concentration values obtained during this study. A correlation matrix revealed that the Mn and Fe oxides/hydroxides are the most important carrier phase for several trace elements, with the exception of rare earth elements (REEs). These also represent a major sink for the observed heavy metal pollution of the soil. REEs are mostly associated with two phases: light (L)REEs are bound to K-Al, while heavy (H)REEs are bound to Mg-bearing minerals. Although there is no direct evidence of a health risk, the paddy soil in the vicinity of Zletovska River needs further investigation and an assessment should be made of its suitability for agricultural use, particularly in view of the highly elevated concentrations of Pb, Zn, As and Cd. PMID- 17120101 TI - Possible linkages between lignite aquifers, pathogenic microbes, and renal pelvic cancer in northwestern Louisiana, USA. AB - In May and September, 2002, 14 private residential drinking water wells, one dewatering well at a lignite mine, eight surface water sites, and lignite from an active coal mine were sampled in five Parishes of northwestern Louisiana, USA. Using a geographic information system (GIS), wells were selected that were likely to draw water that had been in contact with lignite; control wells were located in areas devoid of lignite deposits. Well water samples were analyzed for pH, conductivity, organic compounds, and nutrient and anion concentrations. All samples were further tested for presence of fungi (cultures maintained for up to 28 days and colonies counted and identified microscopically) and for metal and trace element concentration by inductively-coupled plasma mass spectrometry and atomic emission spectrometry. Surface water samples were tested for dissolved oxygen and presence of pathogenic leptospiral bacteria. The Spearman correlation method was used to assess the association between the endpoints for these field/laboratory analyses and incidence of cancer of the renal pelvis (RPC) based on data obtained from the Louisiana Tumor Registry for the five Parishes included in the study. Significant associations were revealed between the cancer rate and the presence in drinking water of organic compounds, the fungi Zygomycetes, the nutrients PO(4) and NH(3), and 13 chemical elements. Presence of human pathogenic leptospires was detected in four out of eight (50%) of the surface water sites sampled. The present study of a stable rural population examined possible linkages between aquifers containing chemically reactive lignite deposits, hydrologic conditions favorable to the leaching and transport of toxic organic compounds from the lignite into the groundwater, possible microbial contamination, and RPC risk. PMID- 17120102 TI - Mobilization of arsenic in groundwater of Bangladesh: evidence from an incubation study. AB - The extensive extraction of arsenic (As)-contaminated groundwaters for drinking, household and agricultural purposes represents a serious health concern in many districts of Bangladesh. This laboratory-based incubation study investigated the sources and mechanisms of As mobilization in these groundwaters. Several incubation studies were carried out using sediments collected from the Bangladesh aquifer that were supplemented, or not, with different nutrients, followed by an analysis of the sediment suspensions for pH, ORP (oxidation-reduction potential), EC (electrical conductivity) and As and Fe(II) concentrations. In the substrate amended sediment suspensions incubated under anaerobic environment, there was a mobilization of As (maximum: 50-67 microg/l) and Fe(II) (maximum: 182 microg/l), while the ORP value decreased immediately and drastically (as much as -468 mV to 560 mV) within 5-6 days. In the sediment suspensions incubated under control and aerobic conditions, no significant As mobilization occurred. The simultaneous mobilization of As and Fe(II) from sediments is a strong indication that their mobilization resulted from the reduction of Fe oxyhydroxide by the enhanced activity of indigenous bacteria present in the sediments; this phenomenon also provides insights on the mobilization mechanism of As in groundwater. The concentrations of As in the sediments used in the incubation studies were strongly linked to the gradients of redox potential development that was stimulated by the quantity of organic nutrient (glucose) used. The penetration of surface-derived organic matter into the shallow aquifer may stimulate the activity of microbial communities, thereby leading to a reduction of iron oxyhydroxide and As release. PMID- 17120103 TI - A methodology for assessing the maximum expected radon flux from soils in northern Latium (central Italy). AB - Northern Latium (Italy) is an area where the Rn risk rate is potentially high because of the extensive outcropping of Neogene U-rich volcanics and the presence of major active tectonic lineaments. The lack of data on Rn risk rates in that area, which is undergoing major urban and industrial development, has prompted this study. It proposes a methodology to evaluate the maximum potential diffusive Rn flux from soils based on the measurement of (226)Ra, (232)Th and (40)K activities by gamma-ray spectrometry, and the measurement of main soil parameters influencing the Rn emanation. This methodology provides a simple, reliable and low-cost tool for drawing up radon flux maps useful to both public planners and private individuals, who want to operate safely in the study area. The proposed methodology may also be applied to other geographic areas outside the prescribed study area. PMID- 17120104 TI - Atmospheric mercury emissions from polluted gold mining areas (Venezuela). AB - Soil, waste rock and mud from mercury-gold amalgamation mining areas of El Callao (Venezuela) are highly enriched in Hg (0.5-500 microg g(-1)) relative to natural background concentrations (<0.1 microg g(-1)). Mercury fluxes to the atmosphere from twelve polluted sites of this area were measured in situ (6 a.m. to 8 p.m.) using a Plexiglas flux chamber connected to a portable mercury analyzer (model RA 915+; Lumex, St. Petersburg, Russia). Mercury fluxes ranged between 0.65 and 420.1 microg m(-2) h(-1), and the average flux range during the diurnal hours was 9.1-239.2 microg m(-2) h(-1). These flux values are five orders of magnitude higher than both reported world background Hg fluxes (1-69 ng m(-2) h(-1)) and the regional values, which are in the range 2-10 ng m(-2) h(-1). The flux results obtained in this study are, however, similar to those measured at Hg polluted sites such as chloro-alkali plants or polymetallic ore mining districts (>100,000 ng m(-2) h(-1)). The results from this study also show that Hg emissions from the soil are influenced by solar radiation, soil temperature and soil Hg concentration. Our data suggest that solar radiation may be the dominant factor affecting Hg degrees emission since the major species of mercury in polluted soil is Hg degrees (85-97% of total Hg). The simple release of Hg degrees vapor is probably the dominant process occurring with incident light in the field. The apparent activation energy for mercury emission indicates that the volatilization of mercury mainly occurred as a result of the vaporization of elemental mercury in soil. The degree of Hg emission differed significantly among the soil sites studied, which may be due to variations in soil texture, organic matter content and soil compaction. PMID- 17120105 TI - Mercury pollution in two typical areas in Guizhou province, China and its neurotoxic effects in the brains of rats fed with local polluted rice. AB - Guizhou province, which located in southwestern of China, is an important mercury (Hg) production center. This study was to investigate the environmental levels and ecological effects of mercury in two typical Hg polluted areas in Guizhou province. In addition, to improve the understanding of the neurotoxic effects of Hg, a rats based laboratory study was also carried out in this study. Samples of water, soil, plants, crops and animals collected from Wanshan mercury mine area, Guzhou province, were analyzed by mercury analyzer. The effects of Hg contaminated rice on the expression of c-jun mRNA in rat's brain and the expression of c-JUN protein in cortex, hippocampus were observed using reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemical methods. The results showed that the mercury contents in most environmental samples of aquatics, soil, atmosphere and the biomass of corn, plant and animals, were higher than the national standard and the corresponding data from unpolluted area. It was found mercury pollutions were significant in soil and air. In the laboratory study, the expression of c-jun mRNA and its protein was significantly induced by Hg polluted rice collected from local area. Selenium could reduce the Hg accumulation in the body and had antagonist effect on Hg in terms of the expression of c-jun mRNA and c-JUN protein. The environmental data and Hg levels in different creatures collected in this study will facilitate the environmental and ecological risk assessment of Hg in the polluted areas. It was urged to be alert of mental health problem in human beings when any kind of Hg-polluted food was taken. More efforts should be performed to protect the local ecosystem and human health in the mercury polluted area of Wanshan, Guizhou province of China. PMID- 17120106 TI - Neuronal nitric oxide synthase immunopositive neurons in cat vestibular complex: a light and electron microscopic study. AB - Nitric oxide is a unique neurotransmitter, which participates in many physiological and pathological processes in the organism. Nevertheless, there are little data about the neuronal nitric oxide synthase immunoreactivity (nNOS-ir) in the vestibular complex of a cat. In this respect, the aims of this study were to: (1) demonstrate nNOS-ir in the neurons and fibers, from all major and accessory vestibular nuclei; (2) describe their light microscopic morphology and distribution; (3) investigate and analyze the ultrastructure of the NOS I immunopositive neurons, fibers, and synaptic boutons. For demonstration of the nNOS-ir, the peroxidase-antiperoxidase-diaminobenzidin method was applied. Immunopositive for nNOS neurons and fibers were present in all major and accessory vestibular nuclei. On the light microscope level, the immunopositive neurons were different in shape and size. According to the latter, they were divided into four groups--small (with diameter less than 15 microm), medium-sized (with diameter from 15 to 30 microm), large type I (with diameter from 30 to 40 microm), and large type II (with diameter greater than 40 microm). On the electron microscope level, the immunoproduct was observed in neurons, dendrites, and terminal boutons. According to the ultrastructural features, the neurons were divided into three groups--small (with diameter less than 15 microm), medium sized (with diameter from 15 to 30 microm), and large (with diameter greater than 30 microm). At least two types of nNOS-ir synaptic boutons were easily distinguished. As a conclusion, we hope that this study will contribute to a better understanding of the functioning of the vestibular complex in cat and that some of the data presented could be extrapolated to other mammals, including human. PMID- 17120107 TI - Chromatin remodeling and repair of DNA double-strand breaks. AB - Eukaryotic cells have developed conserved mechanisms to efficiently sense and repair DNA damage that results from constant chromosomal lesions. DNA repair has to proceed in the context of chromatin, and both histone-modifiers and ATP dependent chromatin remodelers have been implicated in this process. Here, we review the current understanding and new hypotheses on how different chromatin modifying activities function in DNA repair in yeast and metazoan cells. PMID- 17120108 TI - Comparing eye movements recorded by search coil and infrared eye tracking. AB - OBJECTIVE: The performance of a new video-based infrared eye tracker (IR) was compared to the magnetic search coil technique (SC). Since the IR offers interesting possibilities as a diagnostic tool in neuro-ophthalmology, it was investigated whether the new device has overcome shortcomings that were reported from former IR systems. METHODS: Horizontal saccades were recorded using the IR and the SC. The IR allowed eye movement recordings at different sampling rates ranging from 250 Hz to 1000 Hz while the SC recorded at 1000 Hz. RESULTS/CONCLUSIONS: The results show that the IR and the SC were in good agreement and produced similar results. In contrast to other studies, the influence of the sampling rate of the IR was small. The saccade main-sequences did not show significant differences. The latency times observed for both systems were mainly in the short-latency range. PMID- 17120109 TI - Generalized peripheral nerve failure during thoracic spine surgery: a case report. AB - OBJECTIVE: Intra-operative systemic changes impairing peripheral nerve function are not commonly detected with electrophysiology. This case presentation illustrates how somatosensory evoked potential (SSEP) monitoring can detect global changes in peripheral nerve excitability during spine surgery. METHODS: A posterior thoracic spine fixation was performed on a young male with multiple traumatic injuries. Bilateral tibial nerve SSEPs were intraoperatively recorded, along with the right median nerve SSEP for control. RESULTS: A rapid, progressive loss of tibial and median nerve potentials (followed by cortical SSEP loss) occurred 90 min after anaesthetic induction. Oxygenation and fluid volume were adequate throughout the case, despite mean airway resistance being elevated (33 cmH(2)0) and blood pressure being low (80/45 mmHg). Corresponding to the decrease in peripheral nerve responses was a drop in end-tidal CO(2) partial pressure (PaCO(2)) from 37 to 25 mmHg. Approximately, 100 min later, the peripheral and cortically generated SSEPs recovered in 2 of 3 limbs monitored. On emergence from anesthesia it was clear that the patient had bitten and kinked the endotracheal tube thus increasing the airway resistance. Ventilation difficulties were magnified with the patient's prone position. Post-operatively there were no sensorimotor deficits. CONCLUSIONS: Somatosensory evoked potential monitoring during spine surgery can detect uncommon generalized nerve conduction block, and further alert surgical teams to a systemic impairment. This was discovered to result from a compromised endotracheal tube. This can apply in various monitoring situations, as the changes affecting the SSEPs were not related to surgical manipulation. PMID- 17120110 TI - Structural genomics on membrane proteins: comparison of more than 100 GPCRs in 3 expression systems. AB - Production of recombinant receptors has been one of the major bottlenecks in structural biology on G protein-coupled receptors (GPCRs). The MePNet (Membrane Protein Network) was established to overexpress a large number of GPCRs in three major expression systems, based on Escherichia coli, Pichia pastoris and Semliki Forest virus (SFV) vectors. Evaluation by immunodetection demonstrated that 50% of a total of 103 GPCRs were expressed in bacterial inclusion bodies, 94% in yeast cell membranes and 95% in SFV-infected mammalian cells. The expression levels varied from low to high and the various GPCR families and subtypes were analyzed for their expressability in each expression system. More than 60% of the GPCRs were expressed at milligram levels or higher in one or several systems, compatible to structural biology applications. Functional activity was determined by binding assays in yeast and mammalian cells and the correlation between immunodetection and binding activity was analyzed. PMID- 17120111 TI - The logos of the genome: genomes as parts of organisms. PMID- 17120112 TI - "What the patient would have decided": a fundamental problem with the substituted judgment standard. AB - Decision making for incompetent patients is a much-discussed topic in bioethics. According to one influential decision making standard, the substituted judgment standard, the decision that ought to be made for the incompetent patient is the decision the patient would have made, had he or she been competent. Although the merits of this standard have been extensively debated, some important issues have not been sufficiently explored. One fundamental problem is that the substituted judgment standard, as commonly formulated, is indeterminate in content and thus offers the surrogate little or no guidance. What the standard does not specify is just how competent one should imagine the patient to be, and what else one ought to envision about the patient's hypothetical outlook and the circumstances surrounding his or her decision making. The article discusses this problem of underdetermined decision conditions. PMID- 17120113 TI - The practice of health care: wisdom as a model. AB - Reasoning and judgement in health care entail complex responses to problems whose demands typically derive from several areas of specialism at once. We argue that current evidence- or value-based models of health care reasoning, despite their virtues, are insufficient to account for responses to such problems exhaustively. At the same time, we offer reasons for contending that health professionals in fact engage in forms of reasoning of a kind described for millennia under the concept of wisdom. Wisdom traditions refer to forms of deliberation which combine knowledge, reflection and life experience with social, emotional and ethical capacities. Wisdom is key in dealing with problems which are vital to human affairs but lack prescribed solutions. Uncertainty and fluidity must be tolerated in seeking to resolve them. We illustrate the application of wisdom using cases in psychiatry, where non-technical aspects of problems are often prominent and require more systematic analysis than conventional approaches offer, but we argue that our thesis applies throughout the health care field. We argue for the relevance of a threefold model of reasoning to modern health care situations in which multifaceted teamwork and complex settings demand wise judgement. A model based on practical wisdom highlights a triadic process with features activating capacities of the self (professional), other (patient and/or carers and/or colleagues) and aspects of the problem itself. Such a framework could be used to develop current approaches to health care based on case review and experiential learning. PMID- 17120114 TI - Provide expertise or facilitate ethical reflection? A comment on the debate between Cowley and Crosthwaite. PMID- 17120115 TI - Helicase: mystery of progression. AB - Helicases mode of unwinding the nucleic acids and translocation along single stranded nucleic acids is still a subject of great curiosity. Based on the energy transduction and electrophilic interactions, we present a model to explain the mode of action of active helicases. This model considers that both strand separation as well as translocation is active processes fueled by NTP hydrolysis. The model proposes that the translocation appears to involve creeping of helicase over the ssNA lattice rather than inchworm movement. PMID- 17120116 TI - Cryogenic synthesis of molecules of astrobiological interest: catalytic role of cosmic dust analogues. AB - We have studied the effects of the substrate, namely amorphous olivine (MgFeSiO(4)) cosmic dust analogues (CDAs), in synthesis of molecules obtained after 200 keV proton irradiation of formamide (NH(2)COH). Formamide has been deposited on the olivine substrate at 20 K. The abundances of new molecular species formed after an irradiation dose of 12 eV/16 amu in formamide pure (i.e. deposited on an inert silicon substrate) and deposited on CDAs have been compared. Specifically, MgFeSiO(4) amorphous olivine is a selective catalyst preventing formation of NH(3) and CN(-) molecules and changing the relative abundances of NH4(+)OCN(-), CO(2), HNCO, CO. We have shown that the role of CDAs has to be taken into account in experiments simulating processes occurring in astronomical environments. PMID- 17120117 TI - Observed molecular alignment in gaseous streams and possible chiral effects in vortices and in surface scattering. AB - Extensive work in this laboratory has been devoted to the study of intermolecular interactions from scattering experiments, in order to provide ingredients for modelling forces acting in systems involving hydrocarbons, the components of atmospheres, and water. Our detection of aligned oxygen in gaseous streams and further evidence on simple molecules has been extended to benzene and various hydrocarbons. Chiral effects can be seen in the differential scattering of oriented molecules, in particular from surfaces. It is pointed out that it may be of pre-biotical interest that we focus on possible mechanisms for chiral bio stereochemistry of oriented reactants, for example when flowing in atmospheres of rotating bodies, specifically the planet earth, as well as in vortex motions of celestial objects. Molecular dynamics simulations and experimental verifications are in progress. PMID- 17120118 TI - Stromatolites as a resource for novel natural products. PMID- 17120119 TI - S.A.M., the Italian Martian simulation chamber. AB - The Martian Environment Simulator (SAM "Simulatore di Ambiente Marziano") is a interdisciplinary project of Astrobiology done at University of Padua. The research is aimed to the study of the survival of the microorganisms exposed to the "extreme" planetary environment. The facility has been designed in order to simulate Mars' environmental conditions in terms of atmospheric pressure, temperature cycles and UV radiation dose. The bacterial cells, contained into dedicated capsules, will be exposed to thermal cycles simulating diurnal and seasonal Martian cycles. The metabolism of the different biological samples will be analysed at different phases of the experiment, to study their survival and eventual activity of protein synthesis (mortality, mutations and capability of DNA repairing). We describe the experimental facility and provide the perspectives of the biological experiments we will perform in order to provide hints on the possibility of life on Mars either autochthonous or imported from Earth. PMID- 17120120 TI - Studies of biominerals relevant to the search for life on Mars. AB - The evidence of the water erosion on Mars is particularly interesting since present climatic conditions are such that liquid water cannot exist at the surface. But, if water was present on the planet in the past, there may have been life, too. Since the discovery of carbonates on Mars also may have very important implications on the possibility that life developed there, we are studying minerals that can have biotic or abiotic origin: calcite (CaCO(3)) and aragonite, a metastable state of calcite.We have analysed biomineral aragonite, in the form of recent sea shells, as well as crystals of mineral aragonite. Infrared spectroscopy in the 2-25 mum wavelength range reveals that, after thermal processing, the biotic samples have a different spectral behaviour from the abiotic ones. As a result, it is possible to distinguish abiotic mineral aragonite from aragonite of recent biological origin.Obviously, if life existed in the past on the Red Planet, we could expect to find "ancient" biotic carbonates, which should therefore be investigated, in order to search for a way of discriminating them from abiotic minerals. For this reason, at the beginning we have considered samples of crushed fossil shells of aragonite composition. Afterwards, in order to take into account that fossilization processes almost always produce a transformation of metastable form (aragonite) into more stable form (calcite), we also studied samples of mineral calcite and different types of fossils completely transformed into calcite. All these biotic fossil samples show the same spectral behaviour as the fresh biotic material after thermal annealing at 485 degrees C. Instead, the calcite behaves like abiotic aragonite.Furthermore, it is known that seashells and other biominerals are formed through an intimate association of inorganic materials with organic macromolecules. The macromolecules control the nucleation, structure, morphology, crystal orientation and spatial confinement of the inorganic phase: this differentiates biominerals from minerals. Analysing the aragonite or calcite fossils with a Scanning Electron Microscope, we found that the fossilization process did not modify the structure of the biominerals which maintain their microscopic characteristics. Looking at the morphology of fossil biominerals, it is evident that the crystals are arranged in complex architectures compared with the compact structure of the mineral crystals. In conclusion, the properties and structure of the biominerals are different from those of the minerals. The rapid increase of the crystalline structure developed under biotic conditions makes these minerals less resistant to thermal treatments, compared with samples of abiotic origin. This result holds both for recent shells as well as all fossil samples. The spectroscopic behaviour of all analysed calcium carbonates of biotic origin is different from that of the abiotic one. Therefore, the infrared spectroscopy is a valid technique to discern the origin of the samples and a powerful tool for analysing in-situ and "sample-return" Mars missions specimens. Also Optical and Scanning Electron Microscopy can be useful to support this type of studies. PMID- 17120121 TI - Protocell-like microspheres from thermal polyaspartic acid. AB - One of the most prominent amino acids to appear in monomer-generating origin-of life experiments is aspartic acid. Hugo Schiff found in 1897 that aspartic acid polymerizes when heated to form polyaspartylimide which hydrolyzes in basic aqueous solution to form thermal polyaspartic acid which is a branched polypeptide. We recently reported at the ISSOL 2005 Conference that commercially made thermal polyaspartic acid forms microspheres when heated in boiling water and allowed to cool. In a new experiment we heated aspartic acid at 180 degrees C for up to 100 h to form thermal polyaspartylimide which when heated in boiling water without addition of base hydrolyzed to form thermal polyaspartic acid which upon cooling formed microspheres. Thermal polyaspartic acid microspheres appear protocell-like in the sense of being prebiotically plausible lattices or containers that could eventually have been filled with just the right additions of primordial proteins, nucleic acids, lipids, and metabolites so as to constitute protocells capable of undergoing further chemical and biological evolution. Thermal polyaspartic acid microspheres are extremely simple models of protocells that are more amenable to precise quantitative experimental investigation than the proteinoid microspheres of Sidney W. Fox. We present here scanning electron microscope images of such thermal polyaspartic acid microspheres. Figure 1 shows thermal polyaspartic acid microspheres from L: aspartic acid heated at 180 degrees C for 50 h, at a magnification of 3,500x. Figure 2 shows thermal polyaspartic acid microspheres from the same sample at a magnification of 7,000x. The thermal polyaspartic acid microspheres have a diameter of approximately 1 mum These images were viewed with a Hitachi S2460N scanning electron microscope at 20 kV acceleration voltage. Figure 1 Thermal polyaspartic acid microspheres from L: -aspartic acid heated at 180 degrees C for 50 h, at a magnification of 3,500x. Figure 2 Thermal polyaspartic acid microspheres from L: -aspartic acid heated at 180 degrees C for 50 h, at a magnification of 7,000x. PMID- 17120122 TI - Primordia vita. Deconvolution from modern sequences. AB - Evolution of the triplet code is reconstructed on the basis of consensus temporal order of appearance of amino acids. Several important predictions are confirmed by computational sequence analyses. The earliest amino acids, alanine and glycine, have been encoded by GCC and GGC codons, as today. They were succeeded, respectively, by A- and G-series of amino acids, encoded by pyrimidine-central and purine-central codons. The length of the earliest proteins is estimated to be 6-7 residues. The earliest mRNAs were short G+C-rich molecules. These short sequences could have formed hairpins. This is confirmed by analysis of modern prokaryotic mRNA sequences. Predominant size of detected ancient hairpins also corresponds to 6-7 amino acids, as above. Vestiges of last common ancestor can be found in extant proteins in form of entirely conserved short sequences of size six to nine residues present in all or almost all sequenced prokaryotic proteomes (omnipresent motifs). The functions of the topmost conserved octamers are not involved in the basic elementary syntheses. This suggests an initial abiotic supply of amino acids, bases and sugars. PMID- 17120123 TI - The evolution of the genetic code revisited. AB - The evolution of the genetic code in terms of the adoption of new codons has previously been related to the relative thermostability of codon-anticodon interactions such that the most stable interactions have been hypothesised to represent the most ancient coding capacity. This derivation is critically dependent on the accuracy of the experimentally determined stability parameters. A new set of parameters recently determined for B-DNA reveals that the codon anticodon pairs for the codes in non-plant mitochondria on the one hand and prokaryotic and eukaryotic organisms on the other can be unequivocally divided into two classes - the most stable base steps define a common code specified by the first two bases in a codon while the less stable base steps correlate with divergent usage and the adoption of a 3-letter code. This pattern suggests that the fixation of codons for A, G, P, V, S, T, D/E, R may have preceded the divergence of the non-plant mitochondrial line from other organisms. Other variations in the code correlate with the least stable codon-anticodon pairs. PMID- 17120124 TI - Characterizing extrasolar terrestrial planets with reflected, emitted and transmitted spectra. AB - NASA and ESA are planning missions to directly detect and characterize terrestrial planets outside our solar system (nominally NASA-Terrestrial Planet Finder and ESA-DARWIN missions). These missions will provide our first opportunity to spectroscopically study the global characteristics of those planets, and search for signs of habitability and life. We have used spatially and spectrally-resolved models to explore the observational sensitivity to changes in atmospheric and surface properties, and the detectability of surface biosignatures, in the globally averaged spectra and light-curves of the Earth. Atmospheric signatures of Earth-size exoplanets might be detected, in a near future, by stellar occultation as well. Detectability depends on planet's size, atmospheric composition, cloud cover and stellar type. According to our simulations, Earth's land vegetation signature (red-edge) is potentially visible in the disk-averaged spectra, even with cloud cover, and when the signal is averaged over the daily time scale. Marine vegetation is far more difficult to detect. We explored also the detectability of an exo-vegetation responsible for producing a signature that is red-shifted with respect to the Earth vegetation's one. PMID- 17120125 TI - Galileo Avionica's technologies and instruments for planetary exploration. AB - Several missions for planetary exploration, including comets and asteroids, are ongoing or planned by the European Space Agencies: Rosetta, Venus Express, Bepi Colombo, Dawn, Aurora and all Mars Programme (in its past and next missions) are good examples. The satisfaction of the scientific request for the mentioned programmes calls for the development of new instruments and facilities devoted to investigate the body (planet, asteroid or comet) both remotely and by in situ measurements. The paper is an overview of some instruments for remote sensing and in situ planetary exploration already developed or under study by Galileo Avionica Space & Electro-Optics B.U. (in the following shortened as Galileo Avionica) for both the Italian Space Agency (ASI) and for the European Space Agency (ESA). Main technologies and specifications are outlined; for more detailed information please refer to Galileo Avionica's web-site at: http://www.galileoavionica.com . PMID- 17120126 TI - Definitely life but not definitively. AB - Although there have been attempts at a definition of life from many disciplines, none is accepted by all as definitive. Some people believe that it is impossible to define 'life' adequately at the moment. We agree with this point of view on linguistic grounds, examining the different types of definition, the contexts in which they are used and their relative usefulness as aids to arriving at a scientific definition of life. We look at some of the more recent definitions and analyse them in the light of our criteria for a good definition. We argue that since there are so many linguistic and philosophical difficulties with such a definition of life, what is needed is a series of working descriptions, which are suited to the audience and context in which they are used and useful for the intended purpose. We provide some ideas and examples of the forms these may take. PMID- 17120127 TI - Interrupted genes in extremophilic archaea: mechanisms of gene expression in early organisms. AB - Extremophilic Archaea populate biotopes previously considered inaccessible for life. This feature, and the possibility that they are the extant forms of life closest to the last common ancestor, make these organisms excellent candidates for the study of evolution on Earth and stimulate the exobiological research in planets previously considered totally inhospitable. Among the other aspects of the physiology of these organisms, the study of the molecular genetics of extremophilic Archaea can give hints on how the genetic information is transmitted and propagated in ancient forms of life. We review here the expression of interrupted genes in a recently discovered nanoarchaeon and the mechanisms of reprogrammed genetic decoding in Archaea. PMID- 17120128 TI - Early inner solar system impactors: physical properties of comet nuclei and dust particles revisited. AB - During the epoch of early bombardment, terrestrial planets have been heavily impacted by cometary nuclei and cometary dust particles progressively injected in the interplanetary medium. Stardust and Deep Impact missions confirm that the nuclei are porous, loosely consolidated objects, with densities below 1,000 kg m( 3), and that they often release small fragments of ices and dust. Recent numerical simulations of the light scattering properties of cometary dust particles indicate that they are highly porous, most likely fractal, and rich in absorbing organics compounds (with a mixture ratio of e.g. 33 to 60% in mass for comet Hale-Bopp). Taking into account the fact that porous structures survive more easily than compact ones during atmospheric entry, such results reinforce the scenario of the early terrestrial planets enrichment--in organics needed for life to originate--by comets. PMID- 17120129 TI - Prebiotic homochirality as a critical phenomenon. AB - The development of prebiotic homochirality on early-Earth or another planetary platform may be viewed as a critical phenomenon. It is shown, in the context of spatio-temporal polymerization reaction networks, that environmental effects--be they temperature surges or other external disruptions--may destroy any net chirality previously produced. In order to understand the emergence of prebiotic homochirality it is important to model the coupling of polymerization reaction networks to different planetary environments. PMID- 17120130 TI - Design and validation of a medication assessment tool for cancer pain management. AB - OBJECTIVE: A clinical tool to examine prescribing in cancer pain management may provide a means to help establish acceptable standards of adherence to treatment guidelines. The study aim was to design and validate a Medication Assessment Tool for Cancer Pain Management (MAT-CP). SETTING: Hospitals in Northern Norway METHOD: The MAT-CP was designed from guideline criteria based on a previously developed method. The tool was validated by peer review before and during field testing on a study sample of cancer patients experiencing pain. MAIN OUTCOME MEASURE: Perceived relevance, utility, and clarity of individual criteria, and reliability of their application to clinical documentation. Frequency of adherence to agreed definitions of guideline criteria. RESULTS: The final tool comprised 36 criteria covering six different aspects of cancer pain management: (1) pain assessment and information transfer, (2) start of strong opioid therapy; (3) current continuous analgesia; (4) current intermittent analgesia; (5) follow up of therapy, and; (6) other care issues. The tool was tested on 109 cancer patients experiencing pain (57 males), mean (SD) age 60.8 (11.5) years. Guideline adherence overall was 61% (n=1704 applicable criteria). The field-testing informed the modification of the MAT-CP to optimise its clarity and utility when applied to patients' clinical documentation. Good inter- and intra-rater reliability (Cohen's kappa kappa=0.86 and kappa=0.95, respectively) were demonstrated in the application. The preliminary application of the tool during field-testing has highlighted the following for further study: (a) Low adherence <50%) to 14 standards concerning start of opioid treatment and pain therapy follow-up, clinical assessment of risk of gastro-intestinal adverse effects among patients on non-steroidal anti-inflammatory drugs (NSAID), current treatment of breakthrough pain, management of nausea/vomiting; (b) High adherence (>75%) to standards of prescribing of continuous analgesia. CONCLUSION: A clinical tool to examine prescribing in cancer pain management has been designed. Face and content validity have been informed by field-testing. The tool requires further study among palliative care specialists as part of the validation required before it can be recommended for clinical use. PMID- 17120131 TI - Community pharmacists in Australia: barriers to information provision on complementary and alternative medicines. AB - OBJECTIVE: To determine, by surveying Australian community pharmacists, the perceived barriers to the provision of information about complementary and alternative medicines (CAMs) and suggestions for overcoming them. METHOD: Anonymous, self-administered survey sent to a random sample of 701 pharmacists registered in three states of Australia in 2004. MAIN OUTCOME MEASURE: Pharmacists' perceived barriers to the provision of information about CAMs. RESULTS: A total of 344 questionnaires were returned by pharmacists (49% response) of which 211 (30%) were currently practising in community pharmacy. Ninety-five percent of surveyed community pharmacists indicated that they personally received enquires about CAMs, with fewer than 15% reporting they were "very confident" in answering queries about safety, interactions or benefits of CAMs. Frequently used CAM information sources were those from manufacturers and distributors, professional newsletters and journals and textbooks. Pharmacists' perceived barriers to the provision of CAM information included a lack of suitable training (most training was informal), deficiencies in available information sources, a lack of managerial support, the need for regulatory changes, consumer beliefs about CAM safety and time constraints due to competing demands in daily practice. Pharmacists proposed improvements to overcome these barriers including improvements to training. CONCLUSION: There is scope for pharmacy professional organisations and educational institutions to further support pharmacists in their practice through providing information on the best information sources available and training that meets the needs of undergraduate students, pharmacists and other pharmacy staff. There is a need to examine regulatory requirements concerning the provision of product information with CAMs in Australia and to implement mechanisms for increasing consumer awareness of regulatory procedures for these medicines. PMID- 17120132 TI - Development of written information for antiretroviral therapy: comprehension in a Tanzanian population. AB - OBJECTIVE: To design and develop a simple, easily readable patient information leaflet (PIL) for a commonly used antiretroviral (ARV) regimen and to evaluate its readability and acceptability in a Tanzanian population. METHOD: A PIL incorporating simple text and pictograms was designed for the antiretroviral regimen of stavudine, lamivudine and efavirenz. The PIL was designed according to established good design guidelines, modified during a multi-stage iterative testing process and piloted in a South African Xhosa population. The PIL was made available in both English and Kiswahili. Sixty Tanzanian participants who were not taking ARVs were interviewed. They were asked to read the PIL in the language of their choice and were then asked a series of two-part questions; the first part required participants to locate the information in the PIL, after which they were asked to explain the information in their own words. Acceptability was assessed through close-ended questions and open-ended feedback. The influence of selected patient characteristics on comprehension of the PIL was investigated using one-way ANOVA and t-tests for independent samples with a significance level set at 0.05. MAIN OUTCOME MEASURE: Comprehension of the written information in an overall percentage understanding. RESULTS: The overall average percentage comprehension of the PIL was 95%. The target set by the EC guideline that at least 80% of participants correctly locate and understand the information was achieved for 19 of the 20 questions. Five of the six instructions illustrated by pictograms were correctly understood by all participants. The only patient characteristics significantly associated with comprehension were educational level and self-reported ease of reading the PIL. Acceptability of the PIL was high and positive comments were associated with simplicity, good design, easy readability and user-friendliness, the latter enhanced by the inclusion of pictograms. CONCLUSION: The PIL designed for this study was shown to be effective in communicating information about ARVs. Patient characteristics must be taken into account when developing written information, and the final document must be tested for comprehension in the target population. PMID- 17120133 TI - Customers' expectations and satisfaction with a pharmacy not providing advanced cognitive services. AB - OBJECTIVE: To assess customers' expectations and satisfaction with a community pharmacy which does not provide advanced professional cognitive services (APCS). METHOD: A non-validated questionnaire was designed; the questionnaire included items on expectations with pharmacy services and on satisfaction with the existing services. An independent interviewer was appointed to survey people leaving the pharmacy; the questionnaire was administered on two randomly chosen days. RESULTS: The mean age of respondents (n=61) was 46 years. Expectations: The most highly ranked item was "Drug available in the pharmacy" (4.97 on a 5-point Likert scale), followed by "Having queries and questions about health problems answered" (4.84). People found it easy "To seek advice on health problems or about medicines from the pharmacist at the counter" (4.48). The lowest ranked item was "reduced waiting time" (3.13). The overall Satisfaction with pharmacy services was high (4.89). CONCLUSIONS: Customers reported a high level of satisfaction in a pharmacy where advanced cognitive services were not available. However, this should not discourage pharmacists from implementing APCS, because patients' expectations are based only on experiences with current distributive services, which fulfill their perceived needs. PMID- 17120134 TI - Retrospective analysis of medication incidents reported using an on-line reporting system. AB - OBJECTIVE: To examine the types of prescribing, administration and dispensing incidents reported to an on-line incident-reporting scheme and determine the types of healthcare professionals responsible for reporting such incidents. METHOD: Retrospective analysis of medication-related incidents reported to an on line incident-reporting scheme in a large (1000-bed) teaching hospital in the UK. MAIN OUTCOME MEASURES: Frequency and type of incidents, the discipline of the health care professional who reported the incident and the stage in the medication use process (prescribing, dispensing, or administration) at which the incident occurred. RESULTS: Over a 26-month study period, there were 495 medication-related incidents reported, of which 38.6% (191) were classified to be a "near miss". Medication-related incidents were reported most often at the stages of administration (230, 46.5%) and prescribing (192, 38.8%), whilst incidents involving dispensing or supply of medication were reported less often (73, 14.7%). Of all the incidents, pharmacists reported 51.9% (257), nursing staff reported 37.6% (186), and doctors reported 9.1% (45). Cardiovascular (149, 30.1%), central nervous system (106, 21.4%), and antibiotic/anti-infective medication (71, 14.3%) were the most common therapeutic categories associated with reports of medication-related incidents. CONCLUSION: An on-line reporting scheme can be used to monitor medication-related incidents at key stages in the medication-use process in secondary care. The types of incidents reported by health care professionals differ markedly, with fewer medication-related incidents being reported by doctors. Future research should explore the prevailing safety culture amongst the different health care disciplines, and examine the impact that information technology has on the willingness of health care professionals to report adverse incidents. PMID- 17120135 TI - A rice gene activation/knockout mutant resource for high throughput functional genomics. AB - Using transfer DNA (T-DNA) with functions of gene trap and gene knockout and activation tagging, a mutant population containing 55,000 lines was generated. Approximately 81% of this population carries 1-2 T-DNA copies per line, and the retrotransposon Tos17 was mostly inactive in this population during tissue culture. A total of 11,992 flanking sequence tags (FSTs) have been obtained and assigned to the rice genome. T-DNA was preferentially ( approximately 80%) integrated into genic regions. A total of 19,000 FSTs pooled from this and another T-DNA tagged population were analyzed and compared with 18,000 FSTs from a Tos17 tagged population. There was difference in preference for integrations into genic, coding, and flanking regions, as well as repetitive sequences and centromeric regions, between T-DNA and Tos17; however, T-DNA integration was more evenly distributed in the rice genome than Tos17. Our T-DNA contains an enhancer octamer next to the left border, expression of genes within genetics distances of 12.5 kb was enhanced. For example, the normal height of a severe dwarf mutant, with its gibberellin 2-oxidase (GA2ox) gene being activated by T-DNA, was restored upon GA treatment, indicating GA2ox was one of the key enzymes regulating the endogenous level of GA. Our T-DNA also contains a promoterless GUS gene next to the right border. GUS activity screening facilitated identification of genes responsive to various stresses and those regulated temporally and spatially in large scale with high frequency. Our mutant population offers a highly valuable resource for high throughput rice functional analyses using both forward and reverse genetic approaches. PMID- 17120136 TI - Nine 3-ketoacyl-CoA thiolases (KATs) and acetoacetyl-CoA thiolases (ACATs) encoded by five genes in Arabidopsis thaliana are targeted either to peroxisomes or cytosol but not to mitochondria. AB - The sub-cellular location of enzymes of fatty acid beta-oxidation in plants is controversial. In the current debate the role and location of particular thiolases in fatty acid degradation, fatty acid synthesis and isoleucine degradation are important. The aim of this research was to determine the sub cellular location and hence provide information about possible functions of all the putative 3-ketoacyl-CoA thiolases (KAT) and acetoacetyl-CoA thiolases (ACAT) in Arabidopsis. Arabidopsis has three genes predicted to encode KATs, one of which encodes two polypeptides that differ at the N-terminal end. Expression in Arabidopsis cells of cDNAs encoding each of these KATs fused to green fluorescent protein (GFP) at their C-termini showed that three are targeted to peroxisomes while the fourth is apparently cytosolic. The four KATs are also predicted to have mitochondrial targeting sequences, but purified mitochondria were unable to import any of the proteins in vitro. Arabidopsis also has two genes encoding a total of five different putative ACATs. One isoform is targeted to peroxisomes as a fusion with GFP, while the others display no targeting in vivo as GFP fusions, or import into isolated mitochondria. Analysis of gene co-expression clusters in Arabidopsis suggests a role for peroxisomal KAT2 in beta-oxidation, while KAT5 co expresses with genes of the flavonoid biosynthesis pathway and cytosolic ACAT2 clearly co-expresses with genes of the cytosolic mevalonate biosynthesis pathway. We conclude that KATs and ACATs are present in the cytosol and peroxisome, but are not found in mitochondria. The implications for fatty acid beta-oxidation and for isoleucine degradation in mitochondria are discussed. PMID- 17120137 TI - Redescriptions of Monosertum parinum (Dujardin, 1845) and M. mariae (Mettrick, 1958) n. comb. from European passerine birds, with an amended generic diagnosis of Monosertum Bona, 1994 (Cestoda: Dilepididae). AB - The type-species of Monosertum Bona, 1994, M. parinum (Dujardin, 1845), is redescribed on the basis of the neotype designated by Bona (1994). M. mariae (Mettrick, 1958) n. comb. (originally Paricterotaenia mariae Mettrick, 1958) is recorded and redescribed from Erithacus rubecula L. (Passeriformes: Turdidae) in the Kaliningradskaya Oblast', Russia (new geographical record). An amended generic diagnosis of Monosertum is presented. PMID- 17120138 TI - Gartner duct cyst simplified treatment approach. AB - We present two patients with Gartner's duct cyst managed with simple marsupialization and successful long-term follow up. PMID- 17120139 TI - Real-time patient-specific finite element analysis of internal stresses in the soft tissues of a residual limb: a new tool for prosthetic fitting. AB - Fitting of a prosthetic socket is a critical stage in the process of rehabilitation of a trans-tibial amputation (TTA) patient, since a misfit may cause pressure ulcers or a deep tissue injury (DTI: necrosis of the muscle flap under intact skin) in the residual limb. To date, prosthetic fitting typically depends on the subjective skills of the prosthetist, and is not supported by biomedical instrumentation that allows evaluation of the quality of fitting. Specifically, no technology is presently available to provide real-time continuous information on the internal distribution of mechanical stresses in the residual limb during fitting of the prosthesis, or while using it and this severely limits patient evaluations. In this study, a simplified yet clinically oriented patient-specific finite element (FE) model of the residual limb was developed for real-time stress analysis. For this purpose we employed a custom made FE code that continuously calculates internal stresses in the residual limb, based on boundary conditions acquired in real-time from force sensors, located at the limb-prosthesis interface. Validation of the modeling system was accomplished by means of a synthetic phantom of the residual limb, which allowed simultaneous measurements of interface pressures and internal stresses. Human studies were conducted subsequently in five TTA patients. The dimensions of bones and soft tissues were obtained from X-rays of the residual limb of each patient. An indentation test was performed in order to obtain the effective elastic modulus of the soft tissues of the residual limb. Seven force sensors were placed between the residual limb and the prosthetic liner, and subjects walked on a treadmill during analysis. Generally, stresses under the shinbones were approximately threefold higher than stresses at the soft tissues behind the bones. Usage of a thigh corset decreased the stresses in the residual limb during gait by approximately 80%. Also, the stresses calculated during the trial of a subject who complained about pain and discomfort were the highest, confirming that his socket was not adequately fitted. We conclude that real-time patient-specific FE analysis of internal stresses in deep soft tissues of the residual limb in TTA patients is feasible. This method is promising for improving the fitting of prostheses in the clinical setting and for protecting the residual limb from pressure ulcers and DTI. PMID- 17120140 TI - Computation of individual latent variable scores from data with multiple missingness patterns. AB - Latent variable models are used in biological and social sciences to investigate characteristics that are not directly measurable. The generation of individual scores of latent variables can simplify subsequent analyses. However, missing measurements in real data complicate the calculation of scores. Missing observations also result in different latent variable scores having different degrees of accuracy which should be taken into account in subsequent analyses. This manuscript presents a publicly available software tool that addresses both these problems, using as an example a dataset consisting of multiple ratings for ADHD symptomatology in children. The program computes latent variable scores with accompanying accuracy indices, under a 'user-specified' structural equation model, in data with missing data patterns. Since structural equation models encompass factor models, it can also be used for calculating factor scores. The program, documentation and a tutorial, containing worked examples and specimen input and output files, is available at http://statgen.iop.kcl.ac.uk/lsc . PMID- 17120141 TI - Enhancement of emulsifier production by Curvularia lunata in cadmium, zinc and lead presence. AB - The influence of cadmium, zinc and lead on fungal emulsifier synthesis and on the growth of filamentous fungus Curvularia lunata has been studied. Tolerance to heavy metals established for C. lunata was additionally compared with the sensitivity exhibited by strains of Curvularia tuberculata and Paecilomyces marquandii-fungi which do not secrete compounds of emulsifying activity. Although C. lunata, as the only one out of all studied fungi, exhibited the lowest tolerance to heavy metals when grown on a solid medium (in conditions preventing emulsifier synthesis), it manifested the highest tolerance in liquid culture - in conditions allowing exopolymer production. Cadmium, zinc and lead presented in liquid medium up to a concentration of 15 mM had no negative effect on C. lunata growth and stimulated emulsifier synthesis. In the presence of 15 mM of heavy metals, both the emulsifier and 24-h-old growing mycelium exhibited maximum sorption capacities, which were determined as 18.2 +/- 2.67, 156.1 +/- 10.32 mg g(-1) for Cd2+, 22.2 +/- 3.40, 95.2 +/- 14.21 mg g(-1) for Zn2+ and 51.1 +/- 1.85, 230.0 +/- 28.47 mg g(-1) for Pb2+ respectively. The results obtained by us in this work indicate that the emulsifier acts as a protective compound increasing the ability of C. lunata to survive in heavy metal polluted environment. Enhancement of exopolymer synthesis in the presence of Cd2+, Zn2+ and Pb2+ may also suggest, at least to some extent, a metal-specific nature of emulsifier production in C. lunata. Due to accumulation capability and tolerance to heavy metals, C. lunata mycelium surrounded by the emulsifier could be applied for toxic metal removal. PMID- 17120142 TI - The RcnRA (YohLM) system of Escherichia coli: a connection between nickel, cobalt and iron homeostasis. AB - The transporter RcnA has previously been implicated in Ni(II) and Co(II) detoxification in E. coli probably through efflux. Here we demonstrate that the divergently described rcnA and rcnR gene products constitute a link between nickel, cobalt and iron homeostasis. Deletion of the rcnA gene resulted in increased cellular nickel, cobalt and iron concentrations. Expression of rcnA was induced by Ni(II) or Co(II). Overproduction of rcnR inhibited induction of rcnA by metal cations but RcnR did not bind to the rcnA promoter in vitro. When rcnR or fur, the gene of the global repressor of iron homeostasis, was deleted, expression of rcnA was also induced by iron. The promoter region of rcnA was positive in a Fur titration (FURTA) in vivo assay indicative of Fur binding. Thus, rcnA is part of the Fur regulon of E. coli. The implications of a connection between the homoeostasis of closely related transition metals are discussed. PMID- 17120143 TI - Cadmium, zinc and iron interactions in the tissues of bank vole Clethrionomys glareolus after exposure to low and high doses of cadmium chloride. AB - In present study, bank voles Clethrionomys glareolus were peritioneally injected with different doses of cadmium, 0, 1.5, 3.0 mg Cd/kg body mass. Animals were sacrificed on the 21st day after cadmium exposure and the liver and kidney were obtained for cadmium, zinc and iron analysis using atomic absorption spectrometry. Results showed that cadmium had accumulated in the tissues according to dosage and sex. Cadmium affected the survival and body masses of dosed females. Cadmium decreased the iron concentrations in the liver of voles, whereas zinc concentrations increased in both the kidney and liver. PMID- 17120144 TI - Estimation of forward and backward mitral flow using indicator dilution technique: a theoretical feasibility study. AB - A new theoretical algorithm is presented for high-resolution mitral flow determination based on the indicator dilution principle. The algorithm allows forward as well as backward time-dependent mitral flow estimation with a beat-to beat resolution. Indices of normal/subnormal left heart functioning, including total stroke volume (TSV), cardiac output (CO), total ejection fraction (TEF), mitral regurgitation volume (MRV) and mitral regurgitation fraction (MRF), are determined. Knowledge of left atrium and ventricle indicator concentration versus time dependencies and the end systolic left atrium and ventricle volumes are sufficient to determine the mitral flow pattern. However, the non-dimensional index of the total ejection fraction can be calculated on the basis of only the indicator concentration. The algorithm was validated by applying it to blood flows and heart chamber volumes derived from a computer simulation of the cardiovascular circulation. First left heart concentrations versus time data were obtained by determining the distribution over a cardiovascular tract of an ideal indicator, a bolus of which was intravenously injected into one of the arms. Then the backward problem of finding mitral flow was solved. The accuracy of the mitral flow estimation depends on the accuracy of end systolic left atrium and ventricle volume data. The method is applicable over a wide range of aortic regurgitation, up to 20% of cardiac output, suggesting that the algorithm might become a robust technique of non-invasive mitral flow assessment, replacing traditional techniques such as nuclear radiography. PMID- 17120145 TI - Primary infection with cytomegalovirus in ulcerative colitis. PMID- 17120146 TI - Is the time for absorption and oxidation of 13c-octanoate really negligible in the gastric emptying breath test? PMID- 17120147 TI - Esophageal impedance/pH monitoring in pediatric patients: preliminary experience with 50 cases. AB - This paper describes multiple intraluminal impedance (MII) in 50 children with typical and atypical gastroesophageal reflux (GER) symptoms and discusses the possible clinical significance of objective numeric data provided by MII computed analysis. Patients underwent 24-hr pH/MII monitoring. Reflux parameters were analyzed with relation to age and reported symptoms. Nonacidic MII events occurred as frequently as acidic ones. A Pathologic Bolus Exposure Index associated with a normal pH Reflux Index was detected in 26% of our series. Significant correlations were found regarding acid and bolus clearing times and their ratio. We conclude that the low rate of symptom occurrence in the pediatric population represents a limit on MII evaluation. Our study confirmed that nonacid GER is at least as frequent as acid GER. As MII provides interesting objective data that could be used in clinical practice, we suggest further research to define normal ranges in the pediatric population. PMID- 17120148 TI - Plasmapheresis in the treatment of hypertriglyceridemic pancreatitis with ARDS. PMID- 17120150 TI - Use of songs to promote independence in morning greeting routines for young children with autism. AB - This study evaluated the effects of individually composed songs on the independent behaviors of two young children with autism during the morning greeting/entry routine into their inclusive classrooms. A music therapist composed a song for each child related to the steps of the morning greeting routine and taught the children's teachers to sing the songs during the routine. The effects were evaluated using a single subject withdrawal design. The results indicate that the songs, with modifications for one child, assisted the children in entering the classroom, greeting the teacher and/or peers and engaging in play. For one child, the number of peers who greeted him was also measured, and increased when the song was used. PMID- 17120149 TI - Eye movement and visual search: are there elementary abnormalities in autism? AB - Although atypical eye gaze is commonly observed in autism, little is known about underlying oculomotor abnormalities. Our review of visual search and oculomotor systems in the healthy brain suggests that relevant networks may be partially impaired in autism, given regional abnormalities known from neuroimaging. However, direct oculomotor evidence for autism remains limited. This gap is critical since oculomotor abnormalities might play a causal role in functions known to be impaired in autism, such as imitation and joint attention. We integrate our oculomotor review into a developmental approach to language impairment related to nonverbal prerequisites. Oculomotor abnormalities may play a role as a sensorimotor defect at the root of impairments in later developing functional systems, ultimately resulting in sociocommunicative deficits. PMID- 17120151 TI - Functional organization within a neural network trained to update target representations across 3-D saccades. AB - The goal of this study was to understand how neural networks solve the 3-D aspects of updating in the double-saccade task, where subjects make sequential saccades to the remembered locations of two targets. We trained a 3-layer, feed forward neural network, using back-propagation, to calculate the 3-D motor error the second saccade. Network inputs were a 2-D topographic map of the direction of the second target in retinal coordinates, and 3-D vector representations of initial eye orientation and motor error of the first saccade in head-fixed coordinates. The network learned to account for all 3-D aspects of updating. Hidden-layer units (HLUs) showed retinal-coordinate visual receptive fields that were remapped across the first saccade. Two classes of HLUs emerged from the training, one class primarily implementing the linear aspects of updating using vector subtraction, the second class implementing the eye-orientation-dependent, non-linear aspects of updating. These mechanisms interacted at the unit level through gain-field-like input summations, and through the parallel "tweaking" of optimally-tuned HLU contributions to the output that shifted the overall population output vector to the correct second-saccade motor error. These observations may provide clues for the biological implementation of updating. PMID- 17120152 TI - Atrioventricular nodal reentrant tachycardia diagnosed by pacemaker electrograms. AB - Electrograms stored by pacemakers or implantable defibrillators enable the diagnosis of numerous atrial or ventricular arrhythmia episodes. We present the case of a patient implanted with a DDD pacemaker for sinus node dysfunction who experienced episodes of tachycardia. Atrioventricular nodal reentrant tachycardia was diagnosed by real-time analysis of electrograms during and at the onset of the tachycardia episodes. Electrophysiological study confirmed the diagnosis and led to radical treatment of the tachycardia by radiofrequency ablation of the slow pathway. PMID- 17120153 TI - Case report: Placement of a coronary sinus lead in a patient with dextrocardia and situs inversus. AB - Situs inversus with dextrocardia is a congenital condition in which the heart is a mirror image of the anatomically normal heart on the right side. Patients with this condition are increasingly surviving into adulthood and may present with heart failure. Several of such patients may benefit from atriobiventricular pacing which may be technically challenging. This case report describes techniques and equipment used to successfully place a coronary sinus lead in a patient with this congenital condition. PMID- 17120155 TI - God's laboratory: religious rationalities and modernity in Ecuadorian in vitro fertilization. AB - Catholicism is the only major world religion that unequivocally bans the use of in vitro fertilization (IVF). Nevertheless, in Ecuador, Catholic IVF practitioners declare God's dominion over their IVF laboratories and clinics in explaining pregnancy outcomes. My analysis of this routine combination of spiritual and material causal models in Ecuadorian IVF contributes to two ongoing discussions about (1) the tensions between "institutional" and popular forms of Catholic religiosity and (2) the proper boundaries of science in modernity. The Catholic Church's historical and contemporary struggle to determine control of the miraculous has usually been characterized as a conflict between educated clergy and humble peasants. In the case of Ecuadorian IVF, we find, instead, educated elites and middle classes participating in this same contestation with the Church, proclaiming their direct ability to harness the power of God to effect material change on earth. This spiritual power to affect clinical outcomes does not take place just anywhere, but in clinic and lab, disrupting another set of presumptions about modern scientific practice and subjectivity. Like other Ecuadorian elites and middle classes, IVF practitioners are heirs to Enlightenment thought, and experience themselves as modern in their participation in these high-tech endeavors. But their spiritual approach to laboratory rationality does not trouble these IVF practitioners' experience of themselves as moderns, prompting a reevaluation of the narratives of scientific modernity that limit their scope to Europe and North America. PMID- 17120154 TI - Comparative mechanisms of branching morphogenesis in diverse systems. AB - Much progress has been made in recent years toward understanding mechanisms controlling branching morphogenesis, a fundamental aspect of development in a variety of invertebrate and vertebrate organs. To gain a deeper understanding of how branching morphogenesis occurs in the mammary gland, we compare and contrast the cellular and molecular events underlying this process in both invertebrate and vertebrate organs. Thus, in this review, we focus on the common themes that have emerged from such comparative analyses and discuss how they are implemented via a battery of signaling pathways to ensure proper branching morphogenesis in diverse systems. PMID- 17120156 TI - Origin stories revisited: IVF as an anthropological project. PMID- 17120157 TI - Differentiation of recurrent astrocytoma from radiation necrosis: a pilot study with 13N-NH3 PET. AB - Differentiation of posttherapy radiation necrosis from recurrent brain tumor remains a challenging diagnostic problem. The combination of the imaging modalities on the basis of different physiologic mechanisms could improve diagnostic accuracy. The present study assessed the role of (13)N-NH(3) PET in differentiating recurrent cerebral astrocytoma from radiation necrosis. METHODS: Seven patients, who were previously treated with conventional external-beam radiation therapy after surgical resection for cerebral astrocytomas, and showed the enhancing brain lesions on T1-weighted gadiolinium-enhanced MR studies performed in 6 months or above after the radiotherapies, were examined prospectively with (13)N-NH(3) and FDG PET. Five lesions with tumor recurrence and two with radiation necrosis were histologically verified by either surgical resection or stereotactic biopsy. One lesion of radiation necrosis was confirmed clinicoradiologically. RESULTS: In all eight lesions the (13)N-NH(3) PET scans were concordant with the final diagnosis (100%, 8/8). The lesions with recurrent tumor showed moderately to markedly increased (13)N-NH(3) uptake (grade = 4-5). The lesions with radiation necrosis showed absent or less (13)N-NH(3) uptake than surrounding area (grade = 1-2). The FDG PET scans were concordant with the final diagnosis in six of eight lesions (75%, 6/8), and there were one false-negative result and one false-positive result. The diagnostic result of (13)N-NH(3) PET was discordant with FDG PET in two lesions. One lesion with gliosis and radiation necrosis showed slightly increased FDG uptake (grade = 4), but less (13)N-NH(3) uptake (grade = 2). The other lesion with anaplastic astrocytoma showed moderately increased (13)N-NH(3) uptake (grade = 4), but slightly less FDG uptake than surrounding area (grade = 2). CONCLUSIONS: The recurrent astrocytomas showed increased (13)N-NH(3) uptake, and the radiation necrosis showed absent or less (13)N-NH(3) uptake, and (13)N-NH(3) seem superior to (18)F-FDG for this purpose, suggesting that (13)N-NH(3) is a promising tracer for separating radiation necrosis from astrocytoma recurrence. However, the patient population in this study was small. Thus, the further studies are needed to settle this issue. PMID- 17120158 TI - Fractionated stereotactic radiotherapy using gamma unit after hyperbaric oxygenation on recurrent high-grade gliomas. AB - BACKGROUND: To reduce this complication and to enhance the radiation effect to hypoxic cells of high-grade gliomas, the authors performed noninvasive fractionated stereotactic radiotherapy (FSRT) using a Gamma unit combined with hyperbaric oxygen (HBO) therapy for the treatment of recurrent disease. PATIENTS AND METHODS: Twenty-five consecutive patients who had previously received radiotherapy with chemotherapy for recurrent high-grade gliomas, including 14 patients with anaplastic astrocytoma (AA) and 11 with glioblastoma multiforme (GBM), underwent Gamma FSRT immediately after HBO therapy (2.5 atmospheres absolute for 60 min). The Gamma FSRT was repeatedly performed using a relocatable head cast. Median tumor volume was 8.7 cc (range, 1.7-159.3 cc), and the median total radiation dose was 22 Gy (range, 18-27 Gy) to the tumor margin in 8 fractions. RESULTS: Actuarial median survival time after FSRT was 19 months for patients with AA and 11 months for patients with GBM, which was significantly different (P = 0.012, log-rank test). Two patients underwent subsequent second FSRT for regional or remote recurrence. Seven patients (28%) underwent subsequent craniotomies and resections at a mean of 8.4 months after FSRT treatment, and 4 of them had radiation effects without viable cells and remained alive for 50-78 months. CONCLUSION: Gamma FSRT after HBO therapy appears to confer a survival benefit for patients with recurrent high-grade gliomas and warrants further investigation. PMID- 17120159 TI - Epidemiological survey of central nervous system germ cell tumors in Canadian children. AB - OBJECTIVES: To determine the incidence and characteristics of pediatric patients with central nervous system (CNS) germ cell tumors (GCT) in Canada. METHOD: A national retrospective review of hospital charts was done on all patients with CNS GCT diagnosed between 1990 and 2004. Patients had to be under age 18 years at the time of diagnosis of a CNS germ cell tumor and be a resident of Canada. Information extracted included age and year of diagnosis, pathological diagnosis, location of tumor, evidence of disseminated disease at time of diagnosis and biological markers. RESULTS: One hundred and twenty-one cases were identified (83 germinoma; 38 non-germinoma germ cell tumor). The mean annual incidence of CNS GCT was 1.06 per million children (0.7 per million for germinoma; 0.3 per million for NGGCT). Though yearly incidences varied, there was no clear trend to increased incidence. Male predominance was noted (2.4:1 for germinoma; 11:1 for NGGCT). The primary locations were the pineal and suprasellar regions. At the time of diagnosis, disseminated disease was not uncommon (22% germinoma; 32% NGGCT). Beta human gonadotrophin was elevated in the serum, cerebrospinal fluid (CSF) or both in 7% of patients with germinoma and 36% of patients with NGGCT. Elevation of alpha-fetoprotein in serum, CSF or both was seen in 34% of patients with NGGCT. CONCLUSION: The incidence of CNS germ cell tumors in Canadian children is similar to that observed in other Western countries. PMID- 17120160 TI - Silent or minute hemorrhages do probably occur even in congenital supratentorial hemangioblastoma cases. PMID- 17120161 TI - Carbon disulfide-induced alterations of neurofilaments and calpains content in rat spinal cord. AB - To investigate the mechanism of carbon disulfide-induced neuropathy, male Wistar rats were randomly divided into two experimental groups and one control group. The rats in two experimental groups were treated with carbon disulfide by gavage at dosages of 300 and 500 mg/kg/day, respectively, five times per week for 12 weeks. Spinal cords of carbon disulfide-intoxicated rats and their age-matched controls were Triton-extracted and ultracentrifuged to yield a pellet fraction of neurofilament (NF) polymer and a corresponding supernatant fraction. Then, the contents of NF triplet proteins (NF-H, NF-M, NF-L) and two calpain isoforms (m calpain and mu-calpain) in both fractions were determined by immunoblotting. In the meantime, the mRNA levels of NF-H, NF-M, and NF-L in spinal cords were quantified using reverse transcriptase-polymerase chain reaction. Results showed that in the pellet fraction, the contents of three NF subunits in both treated groups decreased significantly except NF-L in low dose group. In the supernatant fraction, the pattern of NFs alteration varied according to dose-levels. Compared to controls, three neurofilmant subunits in the high dose group displayed significant reduction consistently. However, in the low dose group, they remained unaffected. As for calpains, the contents of mu-calpain in both fractions increased significantly regardless of carbon disulfide dose-levels. Meanwhile, m calpain demonstrated a significant decline in the supernatant fraction, and remained unchangeable in the pellet fraction compared to the control group. Furthermore, the levels of mRNA expression of NF-H, NF-M, and NF-L genes were elevated consistently in CS(2)-treated groups. These findings suggested that carbon disulfide intoxication was associated with obvious alterations of NFs content in rat spinal cord, which might be involved in the development of carbon disulfide neurotoxicity. PMID- 17120163 TI - Supportive psychoanalytic psychotherapy for borderline patients: an empirical approach. AB - The author describes a reformulation for research purposes of a supportive psychoanalytic therapy for patients with borderline personality disorder. The major influences on the thinking that led to the modification of prior formulations and the historical background of treatment manuals are described. Supportive treatment is contrasted with transference-focused psychotherapy and dialectic behavior therapy in the interest of creating a clear comparison that lends itself to empirical study. PMID- 17120162 TI - Regulation of phosphorylation of tau by protein kinases in rat brain. AB - Microtubule associated protein tau is abnormally hyperphosphorylated in Alzheimer disease (AD) brain. To investigate the role of protein kinases involved in this lesion, metabolically active slices made from brains of adult rats were treated with or without various specific kinase activators in oxygenated artificial cerebrospinal fluid. The basal kinase activities of protein kinase-A (PKA), CaM Kinase II and GSK-3 were stimulated more than two-fold by isoproterenol, bradykinin and wortmannin, respectively. We found that cdk5 activity was co stimulated with PKA by isoproterenol. Sequential activation of PKA (+cdk5), CaM Kinase II and GSK-3 produced hyperphosphorylation of tau at Ser-198/Ser-199/Ser 202, Ser-214, Thr-231/Ser-235, Ser-262, Ser-396/Ser-404 and Ser-422 sites. Like AD P-tau, the P-tau from brain slices bound to normal tau and its binding to tubulin was inhibited. These studies suggest that PKA, cdk5, CaM Kinase II and GSK-3 are involved in the regulation of phosphorylation of tau and that AD-type phosphorylation of tau is probably a product of the synergistic action of two or more of these kinases. PMID- 17120164 TI - The human serotonin 1A receptor expressed in neuronal cells: toward a native environment for neuronal receptors. AB - 1. The serotonin(1A) (5-HT(1A)) receptor is an important representative of G protein coupled family of receptors. It is the most extensively studied among the serotonin receptors, and appears to be involved in various behavioral and cognitive functions. 2. We report here the pharmacological and functional characterization of the human serotonin(1A) receptor stably expressed in HN2 cell line, which is a hybrid cell line between hippocampal cells and mouse neuroblastoma. 3. Our results show that serotonin(1A) receptors in HN2-5-HT(1A)R cells display ligand-binding properties that closely mimic binding properties observed with native receptors. We further demonstrate that the differential discrimination of G-protein coupling by the specific agonist and antagonist, a hallmark of the native receptor, is maintained for the receptor in HN2-5-HT(1A)R cells. Importantly, the serotonin(1A) receptor in HN2-5-HT(1A)R cells shows efficient downstream signalling by reducing cellular cyclic AMP levels. 4. We conclude that serotonin(1A) receptors expressed in HN2-5-HT(1A)R cells represent a useful model system to study serotonin(1A) receptor biology, and is a potential system for solubilization and purification of the receptor in native-like membrane environment. PMID- 17120165 TI - Irradiation at 634 nm releases nitric oxide from human monocytes. AB - Previous studies have shown that irradiation at 634 nm decreases the release of extracellular reactive oxygen species (ROS) without affecting viability in human monocytes. Here, we examined the effect of irradiation at 634 nm on the release of nitric oxide (NO), activation of inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS), and release of intracellular ROS. Chemiluminescence assays were used to measure NO release, intracellular ROS, and adenosine triphosphate levels (to assess cell viability). Levels of iNOS and eNOS mRNA were analyzed using PCR. Irradiation resulted in elevated levels of NO but had no effect on iNOS or eNOS. Irradiation also caused a decrease in levels of intracellular ROS and had no effect on cell viability. Our studies indicate that irradiation at 634 nm releases NO, possibly from a preformed store, and reduces the production of intracellular ROS without affecting cell viability. Irradiation at 634 nm may have a wide range of clinical applications, including a reduction in oxidative stress-mediated injury in the vasculature. PMID- 17120166 TI - Preliminary trial to investigate temperature of the iPulse intense pulsed light (IPL) glass transmission block during treatment of Fitzpatrick II, IV, V, and VI skin types. AB - The glass transmission block, a key component of all intense pulsed light (IPL) devices, is responsible for the delivery of IPL energy from the xenon discharge lamp to hair and skin structures during treatment. The purpose of this study was to investigate the variation in temperature of the quartz glass block used in the iPulse (CyDen, Swansea, UK) handset during typical hair removal treatments of Asian and Afro-Caribbean skin types. Initial results from four subjects indicated that the temperature of the glass transmission block did not exceed 45 degrees C during any of the treatments. Furthermore, the development of the temperature measurement methodology described in this paper will enable the comparison of data from different IPL systems to be undertaken in a subsequent larger scale trial. PMID- 17120167 TI - Anti-microbial photodynamic therapy: useful in the future? AB - Previous chapters in this volume have focused on fundamental principles and clinical applications of PDT. This chapter will attempt to outline emerging areas of research to identify some new applications that may become useful in the future in clinical practise. The worldwide rise in antibiotic resistance has driven research to the development of novel anti-microbial strategies. Cutaneous diseases caused by MRSA are ideally suited to treatment by anti-microbial photodynamic therapy for eradicating localized infections and for modulating wound healing due to the ability to deliver photosensitizer and light with topical application. The use of photosensitizer and light as an anti-microbial agent against periodontal microbial biofilms should also represent an attractive method of eliminating oral bacteria. Suitable light sources, laser light and non coherent light will be briefly covered. This chapter will focus on some aspects of anti-microbial photodynamic therapy that appear to be promising for dermatological indications and inactivation of pathogenic bacteria within the oral cavity. PMID- 17120168 TI - High-intensity laser for acupuncture-like stimulation. AB - The insertion of needles into specific parts of the body was shown to provide analgesic and therapeutic effects. In this study, we tested the analgesic effects of high-intensity infrared laser for acupuncture-like stimulation. Twelve adult Sprague-Dawley rats weighing 230 to 250 g were randomly assigned to laser, needle, or restraint groups. Stimulation was directed to the meridian point Taixi (KI 3) for 10 min. For laser stimulation, a pulsed Er:YAG system was used. The laser settings were adjusted to provide a focal raise in the skin temperature to about 45 degrees C. The anti-nociceptive effect was evaluated by the tail-flick test. Both needling and laser stimulation significantly increased the tail-flick latency. Peak needling effect was observed immediately after treatment, while laser stimulation was effective both immediately and 45 min after treatment. High intensity laser stimulation may be used alternatively or in combination with conventional acupuncture needling for pain relief. PMID- 17120169 TI - Clinical relevance of urodynamic investigation tests prior to surgical correction of genital prolapse: a literature review. AB - Urodynamic investigations have been advocated to be incorporated in the standard diagnostic workup of patients undergoing surgical correction of genital prolapse. Stress incontinence is reported by 40% of patients with genital prolapse. Urodynamic stress incontinence (USI) is diagnosed in 70-75% of these patients. Occult urodynamic stress incontinence (OUSI) is diagnosed in about 50% of the patients with genital prolapse not reporting stress incontinence before surgery. Performing urodynamic investigation in patients undergoing prolapse surgery may be valuable if diagnosing USI or OUSI results in the selection of the optimal treatment strategy. This treatment strategy is either a combination of prolapse and stress incontinence surgery or prolapse surgery at the beginning and re evaluation of possible stress incontinence afterwards. The combination of prolapse and stress incontinence surgery has the advantage of attempting to solve two problems at the same moment, but carries an increased risk on unwanted side effects, of which, voiding dysfunction and detrusor overactivity are the most important. This review presents a literature overview of what has been proven about the diagnostic and therapeutic value of urodynamic investigations in patients undergoing prolapse surgery. PMID- 17120170 TI - Questionnaires for women with pelvic floor disorders. PMID- 17120171 TI - Monarc transobturator sling system for the treatment of female urinary stress incontinence: results of a post-operative transvaginal ultrasonography. AB - The aim of the study was to determine Monarc (American Medical Systems) sling position after surgical treatment of stress urinary incontinence (SUI) through the transobturator approach. A total of 54 consecutive women with SUI were evaluated post-operatively with transvaginal ultrasound. A concomitant hysterectomy was performed in ten cases and a concomitant prolapse surgery in six cases. Ultrasound measurements include urethral length, the distance between the upper edge of the sling and the bladder neck (BN-S) and the BN-S/U ratio. The mean distance between the transobturator tape and the bladder neck was found to be 12.6 +/- 3.2 mm in the group of patients who underwent the transobturator procedure alone, 13 +/- 3.1 mm in the transobturator plus hysterectomy group and 12 +/- 2.8 mm in the transobturator plus prolapse group. The superior tape margin was at the mid-urethra in 81.5% of patients and always at a distance greater than 7 mm from the bladder neck. Eight patients did not have satisfactory results after the surgery. Only in one out of these eight patients was the transobturator sling not found to be at the mid-urethra. The superior tape margin of the Monarc sling remained at the level of mid-urethra in the majority of cases. It was never located too proximally beneath the bladder neck. PMID- 17120172 TI - A randomized trial of local anesthesia with intravenous sedation vs general anesthesia for the vaginal correction of pelvic organ prolapse. AB - The purpose of this study is to compare the feasibility of local anesthesia with IV sedation versus general anesthesia for vaginal correction of pelvic organ prolapse. Patients with pelvic organ prolapse who were scheduled for an anterior or posterior colporrhaphy, or an obliterative procedure, and who did not have a contraindication or preference to type of anesthesia were randomized to one of the two anesthesia groups. Nineteen patients were randomized to the general group and 21 patients were randomized to the local group. Mean operating room, anesthesia, and surgical time were similar in each group, and 10 patients in the local group bypassed the recovery room. Requests and doses of antiemetics, postoperative verbal numerical pain scores and length of hospital stay were similar between the two groups. Mean recovery room and total hospital costs were significantly lower in the local group. Local anesthesia with IV sedation is a feasible alternative for vaginal surgery to correct pelvic organ prolapse. PMID- 17120173 TI - Bias in research and conflict of interest: why should we care? PMID- 17120174 TI - Do pregnant women exercise their pelvic floor muscles? AB - The aims of the present study were to assess the number of women performing pelvic floor muscle training (PFMT) during pregnancy and to compare the background variables in those exercising and in those who did not. Four hundred and sixty-seven pregnant women (response rate 84%), mean age 31.5 years (range 20 49), answered a questionnaire on general physical activity level during pregnancy including PFMT. The questionnaire was sent out in week 32 of gestation and answered within week 36. Twenty-four percent reported problems with urinary incontinence and 9% flatus/fecal incontinence. The percentages of pregnant women performing PFMT at least once a week before pregnancy and during trimesters 1, 2, and 3 were 7, 12.9, 17.6, and 17.4%, respectively. More women with lower prepregnancy BMI and with present and past pelvic girdle pain were performing regular PFMT. No significant differences were found in any other background variables. It is concluded that relatively few women perform regular PFMT during pregnancy. In conclusion, only 17% of pregnant Norwegian women reported performing PFMT during pregnancy. PMID- 17120175 TI - Prophylactic single-dose prulifloxacin for catheter-associated urinary tract infection after tension-free vaginal tape procedure. AB - The purpose of the study was to evaluate whether antibiotic prophylaxis with a single dose of prulifloxacin after tension-free vaginal tape (TVT) procedure decreases catheter-associated urinary tract infections (UTIs). Patients undergoing TVT procedure receiving a single dose of prulifloxacin (600 mg) 10 h before catheter removal (n=54) were compared with historical controls who have had no treatment (n=60). All patients had an indwelling urethral catheter in situ for 24 h. Urine specimens for culture were collected preoperatively, before Foley removal, and 7 days postoperatively in both groups. Main outcome measures were significant bacteriuria and symptomatic UTI at catheter removal and 1 week postoperatively. No patient had a positive urine culture in the prulifloxacin group compared with 14 (23.3%) in the control group at catheter removal (P=0.0001). Ten out of 14 (71.4%) women with a positive catheter urine culture had a symptomatic UTI. One week after surgery no difference was found in the rate of significant bacteriuria between groups. Our data suggest that a prophylactic single dose of prulifloxacin could help to reduce the rate of symptomatic and asymptomatic UTIs after short-term urethral catheterization in women undergoing TVT procedure. PMID- 17120176 TI - Acute urinary retention caused by a large hydrosalpinx. AB - Urinary retention is a rare condition in women. We report a case of intermittent retention associated with severe anterior displacement of the uterus and cervix caused by a large hydrosalpinx. We also highlight the possibility of a falsely elevated post-void residual as measured by a bladder scan due to hydrosalpinx. Resolution of the patient's symptoms was documented following salpingectomy. PMID- 17120177 TI - Correlation of pelvic organ prolapse quantification system scores with obstetric parameters and lower urinary tract symptoms in primiparae postpartum. AB - This study investigated the correlation between results of the pelvic organ prolapse quantification (POPQ) system at 3 days and at 2 months postpartum with obstetric parameters and lower urinary tract symptoms (LUTS) in 125 primiparae with vaginal delivery. The clinical characteristics, prevalence of pregnancy related LUTS, and POPQ scores were evaluated. Regarding the relationship of obstetric parameters with POPQ scoring, the gh was found positively correlated with the body mass index and vaginal laceration at 2 months postpartum. The POPQ evaluation did not find the LUTS to be significantly related to the prolapse score. The mean scores of points C and D were significantly increased, and gh, pb, and tvl were significantly decreased between the initial and 2-month follow up scores. Our results revealed that a decrease in vaginal size is the principal change during the first 2 months postpartum and that with the exception of gh, neither the obstetric parameters nor the LUTS were associated with the POPQ scoring system. PMID- 17120178 TI - Bone microarchitecture in males with corticosteroid-induced osteoporosis. AB - SUMMARY: Microarchitectural changes in trabecular bone were analyzed by microcomputed tomography (microCT) and histomorphometry in 24 patients with corticosteroid-induced osteoporosis. The microCT images revealed a reduction in trabecular thickness only on frequency distribution curves, with no increase in trabecular separation. Trabecular plate thinning and perforations were easily identified. INTRODUCTION: Corticosteroid-induced osteoporosis (CSIOP) is mediated by direct actions of the drug on bone cells. The result is a decrease in trabecular bone mass and a reduction in trabecular thickness, but connectivity is believed to remain rather well preserved. METHODS: Twenty-four transiliac bone biopsies from patients with CSIOP were studied conjointly by histomorphometry [with two-dimensional (2D) architectural descriptors] and microCT (with 3D analysis of trabecular characteristics, including trabecular thickness and separation). The frequency distribution of thickness and separation were compared with data obtained in nine control subjects. RESULTS: 2D histomorphometry revealed a decrease in bone volume and trabecular thickness in the bone biopsies of the CSIOP patients when compared to those of the controls. MicroCT appeared to be able to identify the reduction in thickness only when the frequency distribution of trabecular thickness was computed. No difference for the curves of the frequency distribution of trabecular separation was evidenced between patients and controls. MicroCT and 2D histomorphometric results were correlated, but 2D analysis appeared to be more sensitive. However, microCT identified a very specific thinning of the trabecular plates in their center that corresponds to the earlier stages of perforations. CONCLUSION: Trabecular plate thinning can be observed and perforations occur on very thin plates in CSIOP patients. PMID- 17120179 TI - Prevalence and predictors of osteoporosis treatment in nursing home residents with known osteoporosis or recent fracture. AB - SUMMARY: We studied nursing home residents with osteoporosis or recent fracture to determine the frequency and predictors of osteoporosis treatment. There was wide variation in performance, and both clinical and systems variables predicted use. This study shows that improvement in osteoporosis care is possible and important for many nursing homes. INTRODUCTION: We determined the prevalence and predictors of osteoporosis evaluation and treatment in high-risk nursing home residents. METHODS: We identified 67 nursing facilities in North Carolina and Arizona with > 10 residents with osteoporosis or recent hip fracture. Medical records (n=895) were abstracted for osteoporosis evaluation [dual-energy X-ray absorptiometry (DXA), vitamin D level, serum calcium), treatment (calcium, vitamin D, osteoporosis medication, hip protectors), clinical, and systems covariates. Data were analyzed at the facility level using mixed models to account for the complex nesting of residents within providers and nursing facilities. RESULTS: Calcium and vitamin D was prescribed for 69% of residents, bisphosphonates for 19%, calcitonin for 14%, other pharmacologic therapies for 6%, and hip protectors for 2%. Overall, 36% received any bone protection (medication or hip protectors), with wide variation among facilities (0-85%). Factors significantly associated with any bone protection included female gender [odds ratio (OR) 2.4, (1.5-3.7)] and nonurban/suburban location [1.5, (1.1-2.2)]. Residents with esophagitis, peptic ulcer disease (PUD), or dysphagia [0.6, (0.4 0.9)] and alcohol abuse [0.2, (0.0-0.9)] were less likely to receive treatment. CONCLUSIONS: There is substantial variation in the quality of osteoporosis treatment across nursing homes. Interventions that improve osteoporosis quality of care are needed. PMID- 17120180 TI - The effect of treatment with a thiazide diuretic for 4 years on bone density in normal postmenopausal women. AB - SUMMARY: We performed a 2-year extension of our previous 2-year randomized controlled trial of the effects of hydrochlorothiazide on bone mineral density. The improvements in bone density seen in the first 2 years were sustained throughout the extension study. Thiazides provide a further option in the prevention of postmenopausal bone loss. INTRODUCTION: Thiazide diuretics reduce urinary calcium excretion and therefore might prevent osteoporosis. Previously we reported a 2-year randomized controlled trial of hydrochlorothiazide treatment in 185 postmenopausal women that showed positive benefits of hydrochlorothiazide on bone density. Here, we report the results of a 2-year extension to that study. METHODS: Of 185 healthy postmenopausal women, 122 agreed to continue in a double blinded 2-year extension taking 50 mg hydrochlorothiazide or placebo daily. Measurements of bone density occurred every 6 months and of calcium metabolism at 2 and 4 years. RESULTS: The improvements in bone density seen in the first 2 years of the trial were sustained throughout the extension. There were significant between-groups differences in the change in bone density over 4 years at the total body (0.9%, P<0.001), legs (1.0%, P=0.002), mid-forearm (1.1%, P=0.03), and ultradistal forearm (1.4%, P=0.04). At the lumbar spine (0.9%, P=0.76) and femoral neck (0.4%, P=0.53) the between-groups differences did not reach statistical significance. CONCLUSIONS: Hydrochlorothiazide produces small positive benefits on cortical bone density that are sustained for at least the first 4 years of treatment. They provide a further option in the prevention of postmenopausal bone loss, especially for women with hypertension or a history of kidney stones. PMID- 17120181 TI - Determinants of bone mineral density in Chinese-American women. AB - SUMMARY: Few data are available regarding bone mineral density (BMD) and its determinants among Chinese Americans. We identified determinants of BMD among 359 Chinese-American women in order to identify risk factors for low BMD in this burgeoning population. BMD in Chinese-American women is influenced by a number of factors, including immigration. INTRODUCTION: Osteoporosis and low BMD are common among Chinese women, including Chinese Americans, who are a growing population at risk for osteoporosis in the US. Few data are available regarding BMD and its determinants among Chinese-American women. METHODS: In this study, we examined predictors of BMD in 359 ambulatory Chinese-American women, ages 20-90, using stepwise multiple regression analysis. Variables in the model included age, weight, height, menarche age, years since menopause, immigration age, years in US, percentage of life in US, number of pregnancies, oral contraceptive use, family history of osteoporosis, family history of hip fracture, daily calcium intake, exercise, time outdoors, alcohol consumption and tobacco use. RESULTS: Among premenopausal women, weight was the strongest predictor of BMD, accounting for 10.5% of the variance at the lumbar spine (LS), 15.2% at the total hip (TH) and 16.6% at the femoral neck (FN). Time outdoors was also a positive predictor of BMD (1.4% at LS, 2.8% at TH and 1.6% at FN), while family history of osteoporosis (1.4% at TH) and age (3.7% at FN) were negative predictors. Among postmenopausal women, greater BMD at the LS and TH was associated with greater weight and earlier immigration age. Weight accounted for 16.4% of the variance at the LS and 19.8% at the TH; immigration age accounted for 3.1% of the variance at the LS and 4.1% at the TH. At the FN, years since menopause and weight were predictors of BMD, accounting for 14.4% and 8.7% of the variance, respectively. While older age at immigration had a negative effect on BMD, years in and proportion of life in the United States were not significant predictors of BMD. CONCLUSIONS: Bone mineral density in Chinese-American women is influenced by a number of biological and lifestyle factors, including immigration. The results of this study provide new insights into risk factors for low bone density as they relate to environmental determinants in the growing population of Chinese American women. PMID- 17120182 TI - A comparative study of the bone-restorative efficacy of anabolic agents in aged ovariectomized rats. AB - INTRODUCTION: The study was designed to compare the bone anabolic effects of basic fibroblast growth factor (bFGF), a selective agonist for prostaglandin E receptor subtype EP4, and parathyroid hormone (PTH) in aged ovariectomized (OVX) rats with severe cancellous osteopenia. METHODS: Groups of aged OVX rats were maintained untreated for 1 year postovariectomy (15 months of age) to develop severe tibial cancellous osteopenia. These animals were then treated with bFGF or the EP4 agonist (EP4) for 3 weeks. Other groups of aged OVX rats were treated with EP4 or PTH alone for 11 weeks, or sequentially with bFGF or EP4 for 3 weeks followed by PTH for 8 weeks. Cancellous and cortical bone histomorphometry were performed in the right proximal tibial metaphysis and tibial diaphysis respectively. RESULTS: Treatment with bFGF for 3 weeks markedly increased serum osteocalcin, osteoid volume, and osteoblast and osteoid surfaces to a greater extent than EP4. Basic FGF, but not EP4 or PTH, induced formation of osteoid islands within bone marrow. EP4 stimulated cancellous bone turnover, but failed to restore lost cancellous bone in the severely osteopenic proximal tibia after 11 weeks of treatment. In contrast, EP4, much like PTH, increased cortical bone mass in the tibial diaphysis by stimulating both periosteal and endocortical bone formation. Treatment of aged OVX rats with PTH alone tended to partially reverse the severe tibial cancellous osteopenia, whereas sequential treatment with bFGF and PTH increased tibial cancellous bone mass to near the level of vehicle treated control rats. These findings indicate that bFGF had the strongest stimulatory effect on cancellous bone formation, and was the only anabolic agent to induce formation of osteoid islands within the bone marrow of the severely osteopenic proximal tibia. Therefore, bFGF may be more effective for the reversal of severe cancellous osteopenia. PTH and EP4 increased cortical bone mass to nearly the same extent, but cancellous bone mass was greater by two-fold in PTH treated OVX rats than in EP4-treated OVX rats. CONCLUSION: These findings in aged OVX rats suggest that PTH is more efficacious than EP4 for augmentation of cancellous bone in the severely osteopenic, estrogen-deplete skeleton. PMID- 17120183 TI - Multiple Z-plasty in pilonidal sinus--a new technique under local anesthesia. AB - BACKGROUND: Pilonidal sinus is a common surgical condition. Though benign, it causes a lot of distress with its associated morbidity and financial loss to the patient. Many procedures have been tried, ranging from nonsurgical to surgical methods (both conservative and radical ones). It is well known that recurrences are low when the scar is away from the midline. MATERIALS AND METHODS: The author has described a new technique of multiple Z-plasty for the treatment of pilonidal sinus that has been effective in the cure of this problem. This article reports a prospective, nonrandomized noncomparative preliminary clinical study of 115 cases of pilonidal sinus operated on by the author over 14 years. RESULTS: The results were compared with the results of other methods mentioned in the literature. There were only two recurrences. CONCLUSIONS: The author proposes that multiple Z plasty can be considered as an option in the surgical treatment of pilonidal sinus. PMID- 17120184 TI - Transient exposure to PTHrP (107-139) exerts anabolic effects through vascular endothelial growth factor receptor 2 in human osteoblastic cells in vitro. AB - Intermittent administration of the N-terminal fragment of parathyroid hormone (PTH) and PTH-related protein (PTHrP) induces bone anabolic effects. However, the effects of the C-terminal domain of PTHrP on bone turnover remain controversial. We examined the putative mechanisms whereby this PTHrP domain can affect osteoblastic differentiation, using human osteosarcoma MG-63 cells and osteoblastic cells from human trabecular bone. Intermittent exposure to PTHrP (107-139), within 10-100 nM, for only 1000 microg/g) at the tumor core at both time points and drug penetration distances of 2.8 and 1.3 mm on day 4 and 8, respectively. Histological examination confirmed necrosis throughout the tumor tissue. Biodegradable polymer millirods successfully treated the primary tumor mass by providing high doxorubicin concentrations to the tumor tissue over an eight day period. PMID- 17120198 TI - Prolonged follow-up after initial therapy with 2-chlorodeoxyadenosine in patients with indolent non-Hodgkin lymphoma: results of Cancer and Leukemia Group B Study 9153. AB - BACKGROUND: The objective of this study was to determine the efficacy and toxicity of 2-chlorodeoxyadenosine (2-CdA) in patients with untreated, indolent non-Hodgkin lymphoma (NHL). METHODS: For this multicenter, single-arm, Phase II study, 44 patients with treatment-naive, stage III or IV, indolent NHL (International Working Formulation subtypes A, B, and C) were enrolled. Patients received 0.14 mg/kg per day of 2-CdA as a 2-hour bolus infusion for 5 consecutive days every 28 days until maximal response or a total of 6 cycles. RESULTS: Thirty eight patients were eligible for response evaluation. The overall response rate was 100% (95% confidence interval [95% CI], 90.8-100%), and the complete response rate was 31.6% (95% CI, 17.5-48.7%). In the intent-to-treat population, the median failure-free survival was 2.0 years (95% CI, 1.3-3.4 years), and the overall survival rate was 7.0 years (95% CI, 4.3-9.4 years). Six patients had sustained remissions that lasted a median of 8.7 years (range, from 5.9 years to > or =11 years). Although 68% of patients experienced at least 1 grade 3 or 4 event, consisting primarily of myelosuppression, severe infections were rare, with only 8 grade 3 infections. Four late malignancies (prostate adenocarcinoma, ductal carcinoma in situ, and myelodysplasia) and 4 patients with large cell transformation were reported. CONCLUSIONS: 2-CdA is an active, well-tolerated therapy for patients with untreated, indolent NHL. PMID- 17120199 TI - Adenosine triphosphate-binding cassette B1 (ABCB1) (multidrug resistance 1) G2677T/A gene polymorphism is associated with high risk of lung cancer. AB - BACKGROUND: P-glycoprotein (P-gp) is a transmembrane transporter that is encoded by the adenosine triphosphate-binding cassette B1 (ABCB1) (multidrug resistance 1) gene, which plays a role in cell defense against environmental attacks, like those generated by xenobiotics. P-gp is expressed in the lung, where it has been suggested to transport these compounds from the interstitium into the lumen. METHODS: Two functional ABCB1 polymorphisms were examined, G2677T/A (in exon 21) and C3435T (in exon 26), in a group of lung cancer patients and in a control group. RESULTS: Whereas 3435T allelic and genotype frequencies were unchanged between both study groups, lung cancer patients showed higher frequency of the 2677T variant allele compared with the control group (0.67 vs. 0.43; P < .001; odds ratio, [OR], 2.6; 95% confidence interval [95% CI], 1.7-4.0). Among the histologic tumor types that were included in the study, squamous cell carcinoma was associated most strongly with the presence of the 2677T allele (OR, 3.92; 95% CI, 2.2-6.9) and especially was associated with the 2677 TT genotype (OR, 6.75; 95% CI, 3.0-15.2). In a haplotype analysis, homozygous wild-type alleles were classified as genotype A, heterozygous alleles were classified as genotype B, and homozygous mutant allele were classified as genotype C both in exon 21 (first letter) and in exon 26 (second letter) loci. The haplotype CB displayed the highest association with lung cancer (OR, 18.09; 95% CI, 2.4-139.2). CONCLUSIONS: The current results taken together suggest that, aside from other known causes of lung cancer, such as tobacco smoke, the existence of polymorphisms in the ABCB1 gene and, specifically, the presence of the G2677T mutation can be crucial in conferring susceptibility to lung cancer. Further studies comprising larger populations are needed to confirm these preliminary findings. PMID- 17120200 TI - Cytotoxicity of partial-stabilized cement. AB - Partial-stabilized cement (PSC) is a kind of modified calcium silicate cement used for root-end surgery. Minor transition metal elements Co, Cr, and Zn were added for enhancing the setting property of to PSC. In our previous study, minor transition metal additions greatly improved the setting property of PSC. However, the concern of metal toxicity was raised, as the material would be used in the human body. In this study, we evaluated the cytotoxicity of PSC in comparison with mineral trioxide aggregate (MTA), which is one of the commercialized materials used for dental root-end filling. Primary osteoblast cell was used as the target cell. Cell proliferation, cytotoxicity, viability, function, and senescence were analyzed. The cytotoxicity of the PSC-Zn group (PSC with Zn addition) was similar to that of MTA. PSC-Zn is not only nontoxic at the cellular level but also has adequate mechanical property, which makes it a potential root end filling material for apical surgery. PMID- 17120201 TI - Increased osteoblast adhesion on nanograined hydroxyapatite and tricalcium phosphate containing calcium titanate. AB - Depending on the coating method utilized and subsequent heat treatments (such as through the use of plasma-spray deposition), inter-diffusion of atomic species across titanium (Ti) and hydroxyapatite (HA) coatings may result. These events may lead to structural and compositional changes that consequently cause unanticipated HA phase transformations which may clearly influence the performance of an orthopedic implant. Thus, the objective of the present in vitro study was to compare the cytocompatibility properties of chemistries that may form at the Ti:HA interface, specifically HA, tricalcium phosphate (TCP), HA doped with Ti, and those containing calcium titanate (CaTiO(3)). In doing so, results of this study showed that osteoblast (bone-forming cells) adhesion increased with greater CaTiO(3) substitutions in either HA or TCP. Specifically, osteoblast adhesion on HA and TCP composites with CaTiO(3) was almost 4.5 times higher than that over pure HA. Material characterization studies revealed that enhanced osteoblast adhesion on these compacts may be due to increasing shrinkage in the unit lattice parameters and decreasing grain size. Although all CaTiO(3) composites exhibited excellent osteoblast adhesion results, Ca(9)HPO(4)(PO(4))(5)OH phase transformation into TCP/CaTiO(3) increased osteoblast adhesion the most; because of these reasons, these materials should be further studied for orthopedic applications. PMID- 17120202 TI - Microscopic analysis of subcutaneous reactions to endodontic sealer implants in rats. AB - The reaction of subcutaneous tissues to Endofill, Endomethasone, Sealer 26, and AH-Plus was investigated microscopically after implantation of in rats polyethylene cannulae, obturated with gutta-percha cones and sealers, in rats. Empty polyethylene cannulae and cannulae filled with gutta-percha cones alone were used as controls. The inflammatory reactions caused by the sealers were evaluated 7, 14, and 30 days after implantation using a descriptive, histopathological analysis. Inflammatory reactions at each implant site were gauged as either absent, discreet, moderate, or intense, and scores from 0 to 3 were attributed, respectively. Microscopic analysis revealed that Endomethasone showed the best biological behavior for all postimplant periods, followed by Sealer 26 and AH Plus, which produced an irritating effect only during the initial pos-implant period. Endofill caused the severest irritation, producing an inflammatory reaction that ranged from moderate to intense over the entire experimental period. Reactions were more intense near those parts of the cannulae containing more sealer. These results reveal that the root canal sealers tested cause inflammation in rat, subcutaneous conjunctive tissue, the intensity of which may be related to the type and quantity of sealer used, and to postimplant period. PMID- 17120203 TI - Synthesis of novel cholic acid functionalized branched oligo/poly(epsilon caprolactone)s for biomedical applications. AB - Novel cholic acid functionalized branched oligo/poly(epsilon-caprolactone)s were synthesized through the ring-opening polymerization of epsilon-caprolactone initiated by cholic acid with hydroxyl groups. The molecular weight of the branched polymers can be adjusted by controlling the feed ratio of the initiator cholic acid to the monomer epsilon-caprolactone. Comparing with linear homopolymer poly(epsilon-caprolactone) (PCL), these branched oligo/poly(epsilon caprolactone)s show much faster hydrolytic degradation rates, implies that our approach provides a convenient and effective strategy to accelerate degradation of the biodegradable polymers with slow degradation rates such as PCL. The cell culture experiment indicates the incorporation of cholic acid moiety to the polymer chain can improve both cell adherence and proliferation obviously. PMID- 17120204 TI - Contrasting effects of collagen and bFGF-2 on neural cell function in degradable synthetic PEG hydrogels. AB - Injectable biodegradable cell carriers provide a potential means to improve transplanted cell viability in the nervous system by providing physical protection from compaction, shear forces, and the acute inflammatory response that occurs following transplantation into the host brain environment. Synthetic polyethylene glycol (PEG) hydrogels are ideal candidates for this purpose, as the degradation profile and mechanical properties of the gel can be controlled. Here we introduce biological components into the synthetic gel with the goal of improving neural cell function in the inert PEG environment. In this study, it was found that (1) bFGF-2 is a survival/mitogenic factor for neural precursor cells in degradable hydrogel cultures, (2) collagen has no measurable effect on cell survival, metabolic activity, or proliferation, and (3) co-application of collagen and bFGF-2 to hydrogel cultures targets cell survival and metabolic activity, an effect that is different than either applied individually. Because collagen and bFGF-2 support the survival and growth of neural cells and other cell types, the co-encapsulation approach and functional characterization described in this study can be extended to the development of an array of tissue engineering applications. These findings suggest the importance of understanding and developing strategies to control the chemical microenvironment surrounding cells in three-dimensional biomaterials. PMID- 17120205 TI - Combined radiofrequency ablation and doxorubicin-eluting polymer implants for liver cancer treatment. AB - Previously, biodegradable polymer implants (polymer millirods) to release chemotherapeutic agents directly into tumors have been developed. The purpose of this study is to evaluate local drug distribution from these implants in liver tumors treated with radiofrequency (RF) ablation and determine if the implants provide a therapeutic improvement over RF ablation alone. Cylindrical implants were fabricated using 65% poly(D,L-lactide-co-glycolide) (PLGA), 21.5% NaCl, and 13.5% doxorubicin. Control or drug-containing millirods were implanted inside VX2 liver tumors (11 mm diameter) in rabbits after RF ablation. Therapeutic efficacy was assessed 4 and 8 days after treatment using tumor size, histology, and fluorescence measurement of drug distribution. Tumors in both test groups recurred at the boundary of the ablated region. Therapeutic doxorubicin concentrations were found in more than 80% of the ablated area, but concentrations declined rapidly at the boundary between normal and ablated tissue. This region was characterized by a developing fibrous capsule with resolving inflammation, which restricted drug transport out of the ablated zone. The intratumoral doxorubicin implants delivered high concentrations of drug within the ablated region but only limited amounts outside the ablation zone. Future studies will focus on overcoming the fibrotic transport barrier and enhancing drug delivery to the periphery of the ablation region to prevent tumor progression. PMID- 17120206 TI - Effects of coumestrol on neonatal and adult mice osteoblasts activities. AB - Estrogen replacement therapy has been shown to reduce postmenopausal osteoporosis. In the present study, we examined the effects of the phytoestrogen coumestrol on neonatal and adult osteoblasts metabolism. Two different sources of osteoblast cells (neonatal mice calvaria and adult mice long bone) cultures were used in this study. The effects of coumestrol on the cellular activities were analyzed by the mitochondrial tetrazolium (MTT) assay, secretion of alkaline phosphatase (ALP), intracellular calcium content (Ca), and the gene expression of bone matrix protein, estrogen receptors (ER-alpha, ER-beta), and osteoprotegerin (OPG) and osteoprotegerin ligand (OPGL). The results showed that the proliferation of neonatal mice osteoblast cells was enhanced by treatment of coumestrol. In the presence of 10(-9)M coumestrol, the osteoblast proliferation attained 139.5% of the control and that the coumestrol can increase the intracellular calcium contents. Type I collagen gene expression was upregulated 167% at the 1st day's culture; ALP gene expression was upregulated 360% at the 7th day's culture; while the osteocalcin gene expression was upregulated 222% at the 14th day's culture. When adult mice osteoblasts were cultured in the presence of 10(-9)M coumestrol, the osteoblasts population increased significantly earlier and attained its maximal effect at the 21st day's culture with 207.4% of control group. The content of ER-beta and osteoprotegerin secretion by neonatal mice control cells gradually increased during osteoblasts differentiation, whereas the ER-alpha and OPGL content were decreased in this study. The cellular responses to the estradiol and counmestrol were quite different in the osteoblasts derived from different age. PMID- 17120207 TI - Bioactive glass as precursor of designed-architecture scaffolds for tissue engineering. AB - In this work, the conditions to obtain concentrated and fluid suspensions from a bioactive glass (55-SiO(2); 41-CaO; 4-P(2)O(5); mol %) were investigated. The influence of the heat treatment of the glass on the specific surface area, solubility, bioactivity, and finally on their dispersion characteristics was studied. Zeta potential and viscosity measurements were carried out, and based on the obtained results, the best dispersant was selected. The optimum concentration of dispersant, maximum content of solid and time of mixing were also investigated. Slurries containing 50 vol % could be obtained calcining the glass at 1100 degrees C and using Darvan 811 (sodium polyacrylate) as dispersant. Scaffolds with designed architecture were prepared from these suspensions combining the gelcasting method and the stereolithography technique. A polymeric negative (replica of the desired structure) was previously obtained by stereolithography. The slurry was cast into the molds and then polymerized (gelcasting method). The negative was eliminated by heat treatment. After sintering at 1300 degrees C, scaffolds with interconnected porosity and three dimensional channels of 400-470 microm and macropores of 1.4 microm were obtained. PMID- 17120208 TI - Fibrinogen adsorption and platelet lysis characterization of fluorinated surface modified polyetherurethanes. AB - A polyetherurethane (PU) was modified using fluorinated surface-modifying macromolecules (SMMs). A double radiolabel method was used simultaneously to measure the number of adhered platelets ((51)Cr) and the quantity of adsorbed Fg ((125)I), in a cone-and-plate instrument. The objectives were to determine if adsorbed Fg levels correlated to platelet adhesion on the surfaces, and to assess if any reductions in platelet adhesion for the SMM-treated surfaces resulted from surface-induced platelet lysis, rather than changes directly related to lower platelet activation and attachment on the novel surfaces. Platelet lysis was determined from lactate dehydrogenase (LDH) and unbound (51)Cr released into plasma isolated from whole blood exposed to test materials. The corresponding Fg adsorption, evaluated under the same platelet adhesion conditions, did not account for the reduced platelet adhesion on the treated surfaces. LDH and (51)Cr platelet release were very low and indicated no statistically significant differences between the materials. It was therefore concluded that platelet lysis did not contribute to the reduction in platelet adhesion characteristic observed on the SMM-treated surfaces. More importantly, the work emphasizes that the platelet activation cannot be inferred to by assessing the quantity of fibrinogen as is commonly done in the literature. The finding suggests a much more complex mechanism of action for the SMM surface modifiers. On-going work is investigating other Fg parameters such as protein binding affinity and protein conformational state in order to establish the mechanism by which the fluorinated surface modifiers may be reducing platelet adhesion via intermediary changes in initial protein adsorption. PMID- 17120209 TI - Molecular mechanisms involved in Ras inactivation: the annexin A6-p120GAP complex. AB - In mammalian cells, a complex network of signaling pathways tightly regulates a variety of cellular processes, such as proliferation and differentiation. New insights from one of the most-important signaling cascades involved in oncogenesis, the Ras-Raf-MAPK pathway, suggest that the subcellular localisation and assembly of signaling modules of this pathway is crucial to control the biological response. This commonly requires membrane targeting events that are mediated by adaptor/scaffold proteins. Of particular interest is the translocation and complex formation of GTPase-activating proteins (GAPs), such as p120GAP, at the plasma membrane to inactivate Ras. Recent studies indicate that one member of the annexin family, annexin A6 acts as a targeting protein for p120GAP. This review discusses how annexin A6 modulates the involvement of negative regulators of the Ras-Raf-MAPK pathway contributing to Ras inactivation. PMID- 17120210 TI - Novel transparent nano- to micro-heterogeneous substrates for in-situ cell migration study. AB - Transparent substrates having heterogeneities ranging from nanometer to micrometer lateral length scale were fabricated to study cell migration. The surfaces were generated using thin films of block copolymers and homopolymer blends on ultra smooth transparent polyethylene terephthalate films. Results show that the lateral size scale of the surface heterogeneities affects fibroblast (NIH-3T3) adhesion, spreading and motility. More specifically, fibroblasts migrate faster on micron-sized than on nanometer-sized heterogeneities. Cell movements and morphology on the micron patterned surfaces resemble cells cultured in a 3D environment. These surfaces, therefore, can potentially be utilized as models to study cell behavior in physiologically relevant conditions which can add to our fundamental understanding of cell-substrate interactions and facilitate development of surfaces for medical devices. PMID- 17120211 TI - Inflammatory response to titanium surfaces with fibrinogen and catalase coatings: an in vitro study. AB - The aim of the present study was to evaluate the possibility to modulate the early inflammatory response in vitro by coating titanium surfaces with candidate proinflammatory (fibrinogen coated turned titanium "Fib") and antiinflammatory proteins (catalase on top of fibrinogen coated turned titanium "Cat"). Additionally, turned titanium surfaces (Ti) were used as controls. The discs were incubated with human mononuclear cells. Adhered cells were investigated with respect to number, viability, differentiation (acute marker 27E10 vs. chronic marker RM3/1), and cytokine production (TNF-alpha and IL-10), after 24 and 72 h. The results indicated that it is possible to modulate the inflammatory response with protein coatings. However, the strongest inflammatory response, indicated by increased number of adhered cells and release of pro and antiinflammatory mediators, was induced by Cat. Furthermore, the cytokine production on this surface was not sensitive to LPS stimulation. Differentiation measured as the expression of the chronic cell surface marker, dominated after 72 h for all surface modifications and Cat displayed an increased number compared to the others. A decrease in the total number of adhered cells and amounts of TNF-alpha were observed on all surfaces over time. The cell viability was, in general, high for all tested surfaces. In conclusion, the study proved it possible to influence the early inflammatory response in vitro by immobilizing protein coatings to titanium surfaces. However, the catalase surface demonstrated the strongest inflammatory response, and the possibility to selectively use the potent antiinflammatory capacity of catalase needs to be further evaluated. PMID- 17120212 TI - Tinkering: a metaphor uniting evolutionary and developmental biology. PMID- 17120213 TI - Chemical profile of adhesive/caries-affected dentin interfaces using Raman microspectroscopy. AB - In clinical practice, dentists must frequently bond adhesives to caries-affected dentin substrates, but the bond that characteristically forms with these substrates does not provide the durability necessary for long-term clinical function. The purpose of this study was to characterize and compare the interfacial chemistry of adhesive with caries-affected and noncarious dentin using micro-Raman spectroscopy. The results indicated that the differences in the Raman spectra between noncarious and caries-affected dentin could not be accounted for by simple decreased mineralization. Both the structure of collagen and mineral in the caries-affected dentin has been altered by the caries process. The differences in structure and composition not only interfered with acid etching process but also subsequent resin monomer penetration. It was shown that the interface between the adhesive and caries-affected dentin was wider and more complicated than that of the adhesive and noncarious dentin. As a result of adhesive phase separation, a structurally integrated hybrid layer did not form at the interface with either caries-affected or noncarious dentin. Using chemical imaging techniques, this study provides the direct evidence of adhesive phase separation at the interface with caries-affected dentin. Although our group previously reported adhesive phase separation at the interface with noncarious dentin, the chemistry of caries-affected dentin leads to greater variability and a more highly irregular composition along the length and breadth of the interface. PMID- 17120214 TI - Distinctin improves the efficacies of glycopeptides and betalactams against staphylococcal biofilm in an experimental model of central venous catheter infection. AB - The ability of microorganisms to adhere to medical implants is a problem of high visibility and has been focused in numerous investigations. To assess the efficacy of distinctin and conventional antibiotics in the treatment of central venous catheter in vitro and in vivo studies were performed. The in vitro susceptibility assay was performed against S. aureus biofilms developed on 96 well polystyrene tissue culture plates. Efficacy studies were performed in a rat model of CVC infection. Twenty-four hours after implantation, the catheters were filled with distinctin. Thirty minutes later, rats were challenged via the CVC with S. aureus. Administration of antibiotics into the CVC at a concentration equal to the MBC for adherent cells, or at 1024 microg/mL began 24 h later. The killing activities of all antibiotics against adherent bacteria were at least four- to eightfold lower than against freely growing cells. When antibiotics were used in distinctin pretreated wells, they showed a significant increase of activity. The in vivo studies showed that when CVCs were pretreated with distinctin biofilm bacterial load was further decreased to 10(1) CFU/mL and bacteremia was not detected. Distinctin displays potential as an adjunctive agent to antibiotics in the treatment of CVC-related infections. PMID- 17120215 TI - Effects of a p38 MAP kinase inhibitor on bone ingrowth and tissue differentiation in rabbit chambers. AB - The effects of an oral p38 mitogen-activated protein kinase (MAPK) inhibitor and polyethylene particles separately and together on tissue differentiation in the bone harvest chamber (BHC) in rabbits over a 3-week treatment period were investigated. The harvested tissue was analyzed histomorphometrically for markers of bone formation (percentage of bone area), osteoblasts (alkaline phosphatase staining), and osteoclasts (CD51, the alpha chain of the vitronectin receptor). Polyethylene particles decreased the percentage of bone ingrowth and staining for alkaline phosphatase. The p38 MAPK inhibitor alone decreased alkaline phosphatase staining. When the oral p38 MAPK inhibitor was given and the chamber contained polyethylene particles, there was a suppression of bone ingrowth and alkaline phosphatase staining. In contrast to oral non-steroidal anti-inflammatory drugs (NSAIDs) and local Interleukin-1 receptor antagonist (IL-1ra) administration, the oral p38 MAPK inhibitor alone did not suppress bone formation when given during the initial phase of tissue differentiation. Particle-induced inflammation and the foreign body reaction were not curtailed when the p38 MAPK inhibitor was given simultaneously with particles. Additional experiments are needed to establish the efficacy of p38 MAPK inhibitor administration on mitigating an established inflammatory and foreign body reaction that parallels the clinical situation more closely. PMID- 17120216 TI - Synthesis, characterization, and in vitro activity of antibiotic releasing polyurethanes to prevent bacterial resistance. AB - Central venous catheters are a major cause of nosocomial bloodstream infections. Different attempts have been made to incorporate antimicrobial agents into catheters, particularly directed at the surface-coating of devices. To facilitate the antimicrobial adsorption, various cationic surfactants, which however showed several problems, have been used. On the other hand, impregnated catheters with only antimicrobials have demonstrated a short-term duration due to the difficulties to deliver the drug slowly. Thus, in order to obtain high antimicrobial-polymer affinity we synthesized or modified polyurethanes to introduce different functional groups. Polymers were loaded with two antibiotics, cefamandole nafate and rifampin (RIF), chosen for both their functional groups and their action spectrum. The in vitro release behavior showed that the elution of drugs depended on the matrix hydrophilicity and on the antibiotic-polymer and antibiotic-antibiotic interactions. To increase the amount of drug released, polyethylene glycol (PEG) used as a pore forming agent at different molecular weights was incorporated in the polymer bulk with antibiotics. As for the in vitro antimicrobial activity of matrices, assessed by Kirby-Bauer test, it was seen that antibiotics released from various formulations inhibited the bacterial growth and exerted a synergistic effect when both were present. In particular, PEG10000-containing polymer was active against the RIF-resistant S. aureus strain up to 23 days. These results suggest that the combined entrapping of antibiotics and pore formers in these novel polymer systems could be promising to prevent the bacterial colonization and to control the emergence of bacterial resistance. PMID- 17120217 TI - Electrospun fibrinogen: feasibility as a tissue engineering scaffold in a rat cell culture model. AB - Fibrinogen has a well-established tissue engineering track record because of its ability to induce improved cellular interaction and scaffold remodeling compared to synthetic scaffolds. While the feasibility of electrospinning fibrinogen scaffolds of submicron diameter fibers and their mechanical properties have been demonstrated, in vitro cellular interaction has not yet been evaluated. The goal of this study was to demonstrate, based on cellular interaction and scaffold remodeling, that electrospun fibrinogen can be used successfully as a tissue engineering scaffold. Electrospun fibrinogen scaffolds were disinfected, seeded with neonatal rat cardiac fibroblasts, and cultured for 2, 7, and 14 days. Cultures were treated to regulate scaffold degradation by either supplementing serum-containing media with aprotinin or crosslinking the scaffolds with glutaraldehyde vapor. Biocompatibility was assessed through a WST-1 cell proliferation assay. Postculture scaffolds were evaluated by scanning electron microscopy and histology. Cell culture demonstrated that fibroblasts readily migrate into and remodel electrospun fibrinogen scaffolds with deposition of native collagen. Supplementation of culture media with different concentrations of aprotinin-modulated scaffold degradation in a predictable fashion, but glutaraldehyde vapor fixation was less reliable. Based on the observed cellular interactions, there is tremendous potential for electrospun fibrinogen as a tissue engineering scaffold. PMID- 17120218 TI - Drug release from pH-responsive thermogelling pentablock copolymers. AB - A novel pH-dependent injectable sustained delivery system was developed by utilizing a cationic pentablock copolymer that exhibits a thermoreversible sol gel transition. Aqueous solutions of the pentablock copolymer, consisting of poly(2-diethylaminoethyl-methyl methacrylate)-poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide)-poly(2-diethylaminoethyl-methyl methacrylate) (PDEAEM(25)-PEO(100)-PPO(65)-PEO(100)-PDEAEM(25)) exhibit temperature and pH dependent micellization due to the lower critical solution temperature of the PPO blocks and the polyelectrolyte character of the PDEAEM blocks, respectively. Aqueous solutions of the copolymers above 12 wt % are free flowing liquids at room temperature and form elastic physical hydrogels reversibly above 37 degrees C. Hydrophobic probe absorbance studies indicate that pentablock copolymer micelles increase the solubility of sparingly soluble drugs. Solutions of the pentablock copolymer that form gels at body temperature exhibit sustained zero order release in in vitro experiments. The release rates of model drugs and proteins were significantly influenced by the pH of the release media, thereby making these polymers ideal candidates for modulated drug delivery. PMID- 17120219 TI - Layer-by-layer assembly of polyelectrolyte films improving cytocompatibility to neural cells. AB - The using of layer-by-layer assembly polyelectrolyte (PE) films has been suggested as a new versatile technique for surface modification aimed at tissue engineering and cell-based chips. In this study, we investigated the surface morphology of the hyaluronic acid (HA)-based PE films deposited on the amino functionalized glass slides using atomic force microscopy. These thin films (bilayer number <9) were measured to have nanoscale roughness ranging from 10 to 100 nm. Then the primary hippocampal and cortical neural cells were cultured on the PE films, respectively. After 5 days of culturing, the cytocompatibility to neural cells was evaluated by cellular morphology, neurite outgrowth, and microtubule-associated protein 2 expressions. From the present results, the HA based PE films were found to be able to support neural cell adhesion and neurite development, especially for the polycation-ending films. It is suggested these HA based multilayer PE films or similar build-ups could thus be used in the future as a way to modify surfaces for nerve scaffolds and neuron-based chips. PMID- 17120220 TI - Thermal oxidation enhances early interactions between human osteoblasts and alumina blasted Ti6Al4V alloy. AB - Oxidation of Ti6Al4V at 500 degrees C for 1 h in air results in the formation of an outer ceramic layer that improves osteoblast behavior and decreases Ti and Al ion release. In this work, alumina blasted Ti6Al4V alloy has been thermally treated and its in vitro biocompatibility has been assessed. Roughness of the blasted alloy was not found significantly altered after heat treatment while chemical surface analysis indicated an increase in stable TiO(2) and Al(2)O(3) oxides. Cell attachment, spreading, cytoskeleton organization as well as cell proliferation, viability, and procollagen I peptide secretion of human primary osteoblasts, impaired on alumina blasted Ti6Al4V, were found to be greatly enhanced on the thermally oxidized blasted alloy. Other informative markers of the osteoblastic phenotype such as alkaline phosphatase, osteocalcin, osteoprotegerin, and mineralized nodule formation were evaluated and indicated that osteoblasts responded at the same extent on untreated and thermally treated blasted alloys. Taken together, our in vitro results indicate that thermal oxidation of alumina blasted Ti6Al4V may favor successful osseointegration by promoting early interactions of osteoblastic cells and the modified surface alloy. PMID- 17120221 TI - In vitro biocompatibility evaluation of poly(3-hydroxybutyrate-co-4 hydroxybutyrate) copolymer in fibroblast cells. AB - Among the various biomaterials available for tissue engineering and therapeutic applications, microbial polyhydroxyalkanoates offer the most diverse range of thermal and mechanical properties. In this study, the biocompatibility of poly(3 hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB); containing 50 mol % of 4 hydroxybutyrate] copolymer produced by Delftia acidovorans was evaluated. The cytotoxicity, mode of cell death, and genotoxicity of P(3HB-co-4HB) extract against V79 and L929 fibroblast cells were assessed using MTT assay, acridine orange/propidium iodide staining, and alkaline comet assay, respectively. Our results demonstrate that P(3HB-co-4HB) treated on both cell lines were comparable with clinically-used Polyglactin 910, where more than 60% of viable cells were observed following 72-h treatment at 200 mg/mL. Further morphological investigation on the mode of cell death showed an increase in apoptotic cells in a time-dependent manner in both cell lines. On the other hand, P(3HB-co-4HB) at 200 mg/mL showed no genotoxic effects as determined by alkaline comet assay following 72-h treatment. In conclusion, our study indicated that P(3HB-co-4HB) compounds showed good biocompatibility in fibroblast cells suggesting that it has potential to be used for future medical applications. PMID- 17120223 TI - Functionalized hydrogel surfaces for the patterning of multiple biomolecules. AB - Patterning of multiple proteins and enzymes onto biocompatible surfaces can provide multiple signals to control cell attachment and growth. Acrylamide-based hydrogels were photo-polymerized in the presence of streptavidin-acrylamide, resulting in planar gel surfaces functionalized with the streptavidin protein. This surface was capable of binding biotin-labeled biomolecules. The proteins fibronectin and laminin, the enzyme alkaline phosphatase, and the photo-protein R phycoerythrin were patterned using soft lithographic techniques. Polydimethylsiloxane stamps were used to transfer biotinylated proteins onto streptavidin-conjugated hydrogel surfaces. Stamped biomolecules were spatially resolved to feature sizes of 10 mum. Fluorescence measurements were used to assess protein transfer and enzyme functionality on modified surfaces. Our results demonstrate that hydrogel surfaces can be patterned with multiple proteins and enzymes, with retention of biological and catalytic activity. These surfaces are biocompatible and provide cues for cell attachment and growth. (c) 2006 Wiley Periodicals, Inc. J Biomed Mater Res 2007. PMID- 17120224 TI - Histological findings of long-term healing of the experimental defects by application of a synthetic biphasic ceramic in rats. AB - Calcium phosphate ceramics are generally biocompatible and can develop interactions with human recipient bone. Therefore, they can be widely used in the field of periodontology and dentistry. The purpose of this investigation was to assess the long-term histological bone healing results of experimentally created critical size parietal bone defects in rats. Twelve Wistar rats were used in this investigation. Two 6-mm wide, symmetrical, and circular critical size defects were created in each parietal bone of the animals. While the right defects filled with granular implant (Ceraform), the symmetrical defects were taken as controls. Eighteen months after implantation, rats were killed and defects including the biomaterial with surrounding bone was taken for histological examination. Serial histological sections were cut across the defects and stained for the histological analysis. Both control and Ceraform implanted regions contained dense collagenous tissue. In the implantation site, multinuclear giant cells were observed around the material. On the other hand, there were no necrosis, tumour, and infection in the implantation region. There was no statistical difference between the control and ceraform implanted groups when the bone formation results were compared (p > 0.05). In conclusion, the results revealed that this material is biocompatible and does enhance the new bone building despite the long-term observation period. Although this biphasic ceramic shows within the limits of the study as a less resorptive and not osteoconductive properties, it can be considered as a biocompatible bone defect filling material having a limited application alternative in dentistry and medicine. PMID- 17120225 TI - Low resilience in suicide attempters: relationship to depressive symptoms. PMID- 17120226 TI - Hooking some stem-group "worms": fossil lophotrochozoans in the Burgess Shale. AB - The fossil record plays a key role in reconstructing deep evolutionary relationships through its documentation of the early diverging stem groups leading to extant phyla. In the middle Cambrian Burgess Shale, two famously problematic worms, Odontogriphus and Wiwaxia, have recently been reinterpreted as stem-group molluscs based on their shared expression of a putative radula and putative ctenidia in Odontogriphus. More detailed analysis of these fossil structures, however, reveals pronounced anatomical and histological discrepancies with molluscan analogues, such that they are more reliably interpreted as primitive features of the superphylum Lophotrochozoa. In the absence of any obviously derived characters, Odontogriphus could be placed in the stem group of the Lophotrochozoa or on the stem of any of its constituent phyla, whereas the dorsal covering of chaetae in Wiwaxia identifies it as a stem-group polychaete. Despite their close relationship, these two jawed, segmented worms could conceivably represent the early stages of two separate phyla. PMID- 17120227 TI - Deeper into the maize: new insights into genomic imprinting in plants. AB - Current models for regulation of parent-specific gene expression in plants have been based on a small number of imprinted genes in Arabidopsis. These present repression as the default state, with expression requiring targeted activation. In general, repression is associated with maintenance methylation of cytosines, while no role has been found in Arabidopsis imprinting for de novo methylation- unlike the case in mammals. A recent paper both reinforces and challenges the model drawn from Arabidopsis. Methylation patterns of two imprinted loci in maize were tracked from gametes to offspring, enabling an exploration of the timing of imprinting. For one gene, fie1, the results were as expected: parent-specific methylation patterns were inherited from the three types of gamete: egg, central cell and sperm. The behaviour of fie2, however, was a surprise: no alleles were methylated in the gametes, although paternally contributed fie2 is methylated and silent in the endosperm, indicating that, in some cases, plant imprinting requires de novo DNA methylation. This work significantly broadens our understanding of plant imprinting and points to a greater diversity in imprinting mechanisms than has previously been appreciated. PMID- 17120228 TI - Serendipitous discovery of novel bacterial methionine aminopeptidase inhibitors. AB - In this article we describe the application of structural biology methods to the discovery of novel potent inhibitors of methionine aminopeptidases. These enzymes are employed by the cells to cleave the N-terminal methionine from nascent peptides and proteins. As this is one of the critical steps in protein maturation, it is very likely that inhibitors of these enzymes may prove useful as novel antibacterial agents. Involvement of crystallography at the very early stages of the inhibitor design process resulted in serendipitous discovery of a new inhibitor class, the pyrazole-diamines. Atomic-resolution structures of several inhibitors bound to the enzyme illuminate a new mode of inhibitor binding. PMID- 17120229 TI - iGibbs: improving Gibbs motif sampler for proteins by sequence clustering and iterative pattern sampling. AB - The motif prediction problem is to predict short, conserved subsequences that are part of a family of sequences, and it is a very important biological problem. Gibbs is one of the first successful motif algorithms and it runs very fast compared with other algorithms, and its search behavior is based on the well studied Gibbs random sampling. However, motif prediction is a very difficult problem and Gibbs may not predict true motifs in some cases. Thus, the authors explored a possibility of improving the prediction accuracy of Gibbs while retaining its fast runtime performance. In this paper, the authors considered Gibbs only for proteins, not for DNA binding sites. The authors have developed iGibbs, an integrated motif search framework for proteins that employs two previous techniques of their own: one for guiding motif search by clustering sequences and another by pattern refinement. These two techniques are combined to a new double clustering approach to guiding motif search. The unique feature of their framework is that users do not have to specify the number of motifs to be predicted when motifs occur in different subsets of the input sequences since it automatically clusters input sequences into clusters and predict motifs from the clusters. Tests on the PROSITE database show that their framework improved the prediction accuracy of Gibbs significantly. Compared with more exhaustive search methods like MEME, iGibbs predicted motifs more accurately and runs one order of magnitude faster. PMID- 17120230 TI - Crystal structure of UDP-N-acetylenolpyruvylglucosamine reductase (MurB) from Thermus caldophilus. PMID- 17120231 TI - Enhanced sampling of peptide and protein conformations using replica exchange simulations with a peptide backbone biasing-potential. AB - During replica exchange molecular dynamics (RexMD) simulations, several replicas of a system are simulated at different temperatures in parallel allowing for exchange between replicas at frequent intervals. This technique allows significantly improved sampling of conformational space and is increasingly being used for structure prediction of peptides and proteins. A drawback of the standard temperature RexMD is the rapid increase of the replica number with increasing system size to cover a desired temperature range. In an effort to limit the number of replicas, a new Hamiltonian-RexMD method has been developed that is specifically designed to enhance the sampling of peptide and protein conformations by applying various levels of a backbone biasing potential for each replica run. The biasing potential lowers the barrier for backbone dihedral transitions and promotes enhanced peptide backbone transitions along the replica coordinate. The application on several peptide cases including in all cases explicit solvent indicates significantly improved conformational sampling when compared with standard MD simulations. This was achieved with a very modest number of 5-7 replicas for each simulation system making it ideally suited for peptide and protein folding simulations as well as refinement of protein model structures in the presence of explicit solvent. PMID- 17120232 TI - Optimization of the concentration of autologous serum for generation of leukemic dendritic cells from acute myeloid leukemic cells for clinical immunotherapy. AB - Clinical application of immunotherapy for acute myeloid leukemia (AML) requires the efficient induction of dendritic cells (DCs) from AML blast cells using in vitro culture. We examined the effect of autologous serum on the properties of leukemic DCs derived from leukemic cells of AML patients by culture in AIM-V medium with GM-CSF, IL-4, TNF-alpha, and 0, 2, 5, or 10% human autologous serum. The expressions of CD80, CD83, CD86, and HLA-DR were upregulated under all culture conditions; however, 10% autologous serum induced the highest expression levels of several molecules. The capacity of leukemic DCs to stimulate allogeneic T cells increased with increasing serum concentration. Stimulation of autologous CD3(+) T cells with leukemic DCs grown in the presence of various concentrations of autologous serum resulted in induction of more IFN-gamma-secreting cells than was the case for unprimed CD3(+) T cells. Leukemic DCs cultured with 10% autologous serum induced the highest numbers of IFN-gamma-secreting cells and CD8(+)CD56(+) T cells from autologous T cells. These results suggest that culture of AML blast cells in the presence of autologous serum could be used to generate leukemic DCs for immunotherapy against AML. The highest serum concentration appeared optimal for generating the most potent leukemic DCs. PMID- 17120233 TI - Effect of immunoadsorption on refractory idiopathic focal and segmental glomerulosclerosis. AB - A case of a young adult with refractory nephrotic syndrome due to focal segmental glomerulosclerosis is reported. Several treatments had been used without success including steroids, cyclophosphamide, cyclosporine A, tacrolimus, and mycophenolate mofetil. Immunoadsorption was performed as a last resort to manage the nephrotic syndrome, which led to a drastic urinary protein reduction. We review the literature supporting immunoadsorption in primary focal segmental glomerulosclerosis. PMID- 17120234 TI - Plasma vitamin values and antiepileptic therapy: case reports of pregnancy outcomes affected by a neural tube defect. AB - BACKGROUND: Folic acid supplementation reduces the occurrence of neural tube defects (NTDs); however, it is not clear whether it protects against teratogenic effects of antiepileptic drugs. METHODS: We report the cases of four pregnant women receiving valproic acid therapy, who all had NTD-affected offspring, despite periconceptional 5 mg/day of folic acid supplementation (cases), and investigated homocysteine metabolism, linked with folate metabolism. Their plasma homocysteine, folates, and vitamin B6 and B12 results were compared with values of two other women, who were also receiving valproic acid and folic acid complement, but who had normal pregnancies (valproic acid controls), and values of 40 pregnant women who had normal pregnancies and were not receiving any therapy (controls without therapy). Because of the possible existence of a genetic susceptibility, polymorphisms in homocysteine metabolism were sought. RESULTS: Two cases showed a decreased phosphopyridoxal level, compared with levels in the controls not receiving therapy. The genotype TT (C677T) is an NTD genetic susceptibility, but it was observed in only one valproic acid control. Various polymorphisms were observed in the cases, but were also common in the controls. Several studies have reported that valproic acid therapy lowers vitamin B6 levels. Our case with the greatest decrease in plasma phosphopyridoxal, who was taking periconceptional folic acid plus pyridoxine therapy, had a normal second pregnancy outcome. CONCLUSIONS: In addition to folates, other vitamins, such as vitamin B6, may have played a role in NTDs in our patients taking an antiepileptic drug. PMID- 17120235 TI - Triphalangeal thumb in association with split hand/foot: a phenotypic marker for SHFM3? AB - BACKGROUND: At least five distinct loci have been implicated in split hand foot malformation (SHFM). Establishing genotype/phenotype correlations at the chromosomal level may elucidate responsible developmental genes and improve patient management. In our analysis of previously published genetically mapped SHFM cases, preaxial hand involvement was a significant discriminating variable, most commonly seen at the SHFM3 locus (OMIM 600095) at 10q24. Of the 47 SHFM3 patients analyzed, 15 (31.9%) had triphalangeal thumb (TPT), a limb finding not reported at any other locus. METHODS: The association of TPT/split foot, in particular, prompted us to review the literature for similar cases. RESULTS: We ascertained a number of unmapped familial and sporadic cases with TPT/split foot, including a group of patients with triphalangeal thumb-brachyectrodactyly syndrome. Certain trends were similar in both SHFM3 and these unmapped literature cases. With respect to gender, 7/12 (58%) of mapped SHFM3 cases with TPT/split foot were male whereas 5/12 (42%) were female, compared with 22/50 (44%) males and 28/50 (56%) females among unmapped cases (P=0.3715). Individuals in both groups usually had bilateral involvement, with 67 and 60% showing bilateral TPT among mapped and literature cases, respectively (P=0.6714). Bilateral involvement of the feet was even more striking (83% of SHFM3 patients and 96% of literature cases; P=0.0808). CONCLUSIONS: Patients with TPT/split foot may in fact represent SHFM3 cases and should be evaluated for genomic rearrangements at 10q24. TPT may be identified only by radiographic analysis, emphasizing the importance of imaging these patients and their family members. PMID- 17120236 TI - Association of paternal age with prevalence of selected birth defects. AB - BACKGROUND: Unlike maternal age, the effect of paternal age on birth defect prevalence has not been well examined. We used cases from the Texas birth defect registry, born during 1996-2002, to evaluate the association of paternal age with the prevalence of selected structural birth defects. METHODS: Poisson regression was used to calculate prevalence ratios (PRs) and 95% confidence intervals (CIs) associated with paternal age for each birth defect, adjusting for maternal age, race/ethnicity, and parity. RESULTS: Relative to fathers ages 25-29 years, fathers 20-24 years of age were more likely to have offspring with gastroschisis (PR 1.47, 95% CI: 1.12-1.94), and fathers 40+ years old were less likely to have offspring with trisomy 13 (PR 0.40, 95% CI: 0.16-0.96). No association was seen between paternal age and prevalence of anencephaly and encephalocele. A selection bias was observed for the other birth defects in which cases of younger fathers were more often excluded from study. CONCLUSIONS: In studies of birth defect risk and paternal age, the source of information may affect the validity of findings. PMID- 17120237 TI - Virus-PLoc: a fusion classifier for predicting the subcellular localization of viral proteins within host and virus-infected cells. AB - Viruses can reproduce their progenies only within a host cell, and their actions depend both on its destructive tendencies toward a specific host cell and on environmental conditions. Therefore, knowledge of the subcellular localization of viral proteins in a host cell or virus-infected cell is very useful for in-depth studying of their functions and mechanisms as well as designing antiviral drugs. An analysis on the Swiss-Prot database (version 50.0, released on May 30, 2006) indicates that only 23.5% of viral protein entries are annotated for their subcellular locations in this regard. As for the gene ontology database, the corresponding percentage is 23.8%. Such a gap calls for the development of high throughput tools for timely annotating the localization of viral proteins within host and virus-infected cells. In this article, a predictor called "Virus-PLoc" has been developed that is featured by fusing many basic classifiers with each engineered according to the K-nearest neighbor rule. The overall jackknife success rate obtained by Virus-PLoc in identifying the subcellular compartments of viral proteins was 80% for a benchmark dataset in which none of proteins has more than 25% sequence identity to any other in a same location site. Virus-PLoc will be freely available as a web-server at http://202.120.37.186/bioinf/virus for the public usage. Furthermore, Virus-PLoc has been used to provide large scale predictions of all viral protein entries in Swiss-Prot database that do not have subcellular location annotations or are annotated as being uncertain. The results thus obtained have been deposited in a downloadable file prepared with Microsoft Excel and named "Tab_Virus-PLoc.xls." This file is available at the same website and will be updated twice a year to include the new entries of viral proteins and reflect the continuous development of Virus-PLoc. PMID- 17120238 TI - Exploratory synthesis of peptide-alpha-thioester segments spanning the polypeptide sequence of the delta-opioid receptor, a G protein-coupled receptor. AB - We have decided to use the delta-opioid receptor (372 residues) as a model system to develop methods for the total chemical synthesis of G protein-coupled receptors. The most important feature of this receptor from a chemical synthesis perspective is the wealth of cysteines spread throughout its sequence, which are required for native chemical ligation. A total of 13 cysteines are located in the the delta-opioid receptor polypetide chain in both loop and putative transmembrane (TM) regions. We envisioned a synthesis of the polypeptide that would make use of peptide-alpha-thioesters ranging from 37 to 63 residues in length. Here, we report data from an exploratory synthesis of such a set of peptide-alpha-thioesters. For all seven peptides, the crude material approximately 30 residues into the synthesis was sufficiently homogeneous to make isolation and purification straightforward. Extension of the peptides to between 40 and 50 residues in length generally produced a significant decrease in the quality of the crude products, although in most cases, we judged that high purity peptides could probably be isolated. By 60 residues, however, the crude peptide product mixtures are probably too heterogeneous to purify to homogeneity by reversed-phase HPLC. In general, delta-opioid receptor peptides with a single predicted TM domain were sufficiently soluble to handle postcleavage and to analyze by reversed-phase HPLC, whereas 1.5 TM domains rendered the peptides too hydrophobic to handle or analyze by standard protocols. Given the challenges of chain assembly, handling, and purification of these peptides, a synthetic strategy that uses approximately 12 or 13 shorter peptide segments of 20-40 residues each is probably a more feasible approach. PMID- 17120239 TI - Managing hepatosplenic gammadelta T-cell leukemia-lymphoma in children. PMID- 17120240 TI - Pharmacokinetic evaluation of darbepoetin alfa for the treatment of pediatric patients with chemotherapy-induced anemia. AB - BACKGROUND: Cancer patients undergoing chemotherapy often develop anemia, which can increase the risk for transfusions and fatigue. The recombinant erythropoiesis-stimulating agent darbepoetin alfa can effectively treat chemotherapy-induced anemia (CIA) in adults, but limited data are available regarding its use in pediatric cancer patients. The goals of this phase 1, open label, uncontrolled study were to assess the pharmacokinetic profile and safety of darbepoetin alfa in pediatric patients with CIA. PROCEDURE: Pediatric patients with nonmyeloid malignancies and CIA received up to six doses of darbepoetin alfa 2.25 mcg/kg subcutaneously. After the first dose, the pharmacokinetic properties of darbepoetin alfa were assessed during a 14-day sampling period. All subsequent doses were given weekly with predose blood samples collected before study drug administration. RESULTS: After a single dose of darbepoetin alfa, the mean (SD) peak serum concentration was 10.5 (3) ng/ml, and the median time to peak concentration was 71.4 hr. Darbepoetin alfa exhibited a mean (SD) terminal half life of 49.4 (32) hr. Upon repeated weekly administration, no evidence of darbepoetin alfa accumulation was observed though there was high intra- and inter individual variability. In addition, darbepoetin alfa was well tolerated; some study patients experienced increases in hemoglobin. CONCLUSIONS: The pharmacokinetic profile of darbepoetin alfa indicated that it was slowly absorbed and exhibited a long terminal half-life in these pediatric study patients with CIA. PMID- 17120241 TI - Outcome of retinoblastoma in east Africa. AB - We estimated the proportion of patients reaching a pediatric ophthalmology unit (Comprehensive Community Based Rehabilitation for Tanzania Disability Hospital, CCBRT) or an oncology unit (ORCI) in east Africa and investigated presentation, histology, and treatment outcomes of patients with retinoblastoma. A 5-year retrospective study identified 91 patients, representing approximately 18% of the nationwide total. Mean lag time was 10 months (standard deviation (SD) = 17) and mean follow-up was 8 months (SD = 11, range 0-40, n = 91). Thirty months disease free survival probability was 0.23 (standard error = 0.07). Outcomes for retinoblastoma in Africa remain poor. The data presented here suggest strategies for improving the outcomes, including encouraging earlier presentation and establishment of multi-disciplinary treatment centers. PMID- 17120243 TI - Simultaneous occurrence of Wilms tumor and Burkitt lymphoma in two brothers. PMID- 17120244 TI - Glutamate activates c-fos in glial cells via a novel mechanism involving the glutamate receptor subtype mGlu5 and the transcriptional repressor DREAM. AB - Activation of c-fos in brain is related to coupling of neuronal activity to gene expression, but also to pathological conditions such as seizures or excitotoxicity-induced cell death. Glutamate activates c-fos in neurons through the calcium-dependent phosphorylation of CREB by ERK and/or CaMKIV kinase pathways downstream NMDA-receptors. In glial cells, however, the activation of c fos by glutamate is poorly understood. Because glial cells actively modulate neuronal excitability and the brain's response to injury, we studied the mechanisms by which glutamate activates c-fos in rat cortical glial cells. Glutamate potently induced c-fos mRNA in a calcium-dependent manner, as demonstrated by using the calcium chelator BAPTA-AM. Glutamate-induced c-fos mRNA expression was not sensitive to inhibitors of ERK, p38(MAPK), or CaMK pathways, indicating that glial c-fos is activated by a distinct mechanism. Thapsigargin abolished the glutamate effect on c-fos mRNA, indicating ER calcium mobilization. Additionally, glutamate induction of c-fos mRNA was sensitive to the mGluR5 antagonist MPEP but not the NMDA-R antagonist MK-801. In luciferase reporter assays, DRE, which actively represses c-fos by binding the calcium-binding transcriptional repressor DREAM, was activated by glutamate, whereas SRE and CRE were not. Finally, glutamate caused the nuclear export of DREAM in astrocytes, and transfection of astrocytes with a mutant variant of DREAM that constitutively binds DNA inhibited glutamate-induced c-Fos expression. These findings are in sharp contrast to the mechanism described in neurons and suggest a novel pathway activated by glutamate in glial cells that employs mGluR5, ER calcium, and the derepression of c-fos at the DRE. PMID- 17120245 TI - Nosology and classification of genetic skeletal disorders: 2006 revision. AB - The objective of the paper is to provide the revision of the Nosology of Constitutional Disorders of Bone that incorporates newly recognized disorders and reflects new molecular and pathogenetic concepts. Criteria for inclusion of disorders were (1) significant skeletal involvement corresponding to the definition of skeletal dysplasias, metabolic bone disorders, dysostoses, and skeletal malformation and/or reduction syndromes, (2) publication and/or MIM listing, (3) genetic basis proven or very likely, and (4) nosologic autonomy confirmed by molecular or linkage analysis and/or distinctive diagnostic features and observation in multiple individuals or families. Three hundred seventy-two different conditions were included and placed in 37 groups defined by molecular, biochemical and/or radiographic criteria. Of these conditions, 215 were associated with one or more of 140 different genes. Nosologic status was classified as final (mutations or locus identified), probable (pedigree evidence), or bona fide (multiple observations and clear diagnostic criteria, but no pedigree or locus evidence yet). The number of recognized genetic disorders with a significant skeletal component is growing and the distinction between dysplasias, metabolic bone disorders, dysostoses, and malformation syndromes is blurring. For classification purposes, pathogenetic and molecular criteria are integrating with morphological ones but disorders are still identified by clinical features and radiographic appearance. Molecular evidence leads to confirmation of individual entities and to the constitution of new groups, but also allows for delineation of related but distinct entities and indicates a previously unexpected heterogeneity of molecular mechanisms; thus, molecular evidence does not necessarily simplify the Nosology, and a further increase in the number of entities and growing complexity is expected. By providing an updated overview of recognized entities with skeletal involvement and of the underlying gene defects, the new Nosology can provide practical diagnostic help, facilitate the recognition of new entities, and foster and direct research in skeletal biology and genetic disorders. PMID- 17120247 TI - Genotype-phenotype correlations in tuberous sclerosis: who and how to treat. PMID- 17120248 TI - Unusually mild tuberous sclerosis phenotype is associated with TSC2 R905Q mutation. AB - OBJECTIVE: To report the clinical manifestations and functional aspects of Tuberous Sclerosis Complex (TSC), resulting from Codon 905 mutations in TSC2 gene. METHODS: We performed a detailed study of the TSC phenotype and genotype in a large French-Canadian kindred (Family A). Subsequently, clinical and molecular data on 18 additional TSC families with missense mutations at the same codon of TSC2 were collected. Functional studies were performed on the different missense changes and related to the phenotype. RESULTS: A 2714G>A (R905Q) mutation was identified in Family A. The TSC phenotype in this family was unusually mild and characterized by hypomelanotic macules or focal seizures that remitted spontaneously or were easily controlled with medication. Diagnostic criteria were met in only a minority of mutation carriers. Other families with the R905Q mutation were found to have a similar mild phenotype. In contrast, patients with a 2713C>T (R905W) or a 2713C>G (R905G) mutation had more severe phenotypes. Although all three amino acid substitutions were pathogenic, the R905W and R905G substitutions affected tuberin function more severely than R905Q. INTERPRETATION: Codon 905 missense mutations in TSC2 are relatively common. The TSC2 R905Q mutation is associated with unusually mild disease, consistent with functional studies. Combined with previous reports, it is apparent that certain TSC2 missense mutations are associated with a mild form of tuberous sclerosis, which in many patients does not meet standard diagnostic criteria. These findings have implications for the large number of patients with limited clinical features of TSC and for genetic counseling in these families. PMID- 17120249 TI - Localization of LRRK2 to membranous and vesicular structures in mammalian brain. AB - OBJECTIVE: The PARK8 gene responsible for late-onset autosomal dominant Parkinson's disease encodes a large novel protein of unknown biological function termed leucine-rich repeat kinase 2 (LRRK2). The studies herein explore the localization of LRRK2 in the mammalian brain. METHODS: Polyclonal antibodies generated against the amino or carboxy termini of LRRK2 were used to examine the biochemical, subcellular, and immunohistochemical distribution of LRRK2. RESULTS: LRRK2 is detected in rat brain as an approximate 280kDa protein by Western blot analysis. Subcellular fractionation demonstrates the presence of LRRK2 in microsomal, synaptic vesicle-enriched and synaptosomal cytosolic fractions from rat brain, as well as the mitochondrial outer membrane. Immunohistochemical analysis of rat and human brain tissue and primary rat cortical neurons, with LRRK2-specific antibodies, shows widespread neuronal-specific labeling localized exclusively to punctate structures within perikarya, dendrites, and axons. Confocal colocalization analysis of primary cortical neurons shows partial yet significant overlap of LRRK2 immunoreactivity with markers specific for mitochondria and lysosomes. Furthermore, ultrastructural analysis in rodent basal ganglia detects LRRK2 immunoreactivity associated with membranous and vesicular intracellular structures, including lysosomes, endosomes, transport vesicles, and mitochondria. INTERPRETATION: The association of LRRK2 with a variety of membrane and vesicular structures, membrane-bound organelles, and microtubules suggests an affinity of LRRK2 for lipids or lipid-associated proteins and may suggest a potential role in the biogenesis and/or regulation of vesicular and membranous intracellular structures within the mammalian brain. PMID- 17120251 TI - Is penumbral imaging useful for extending the treatment window for intravenous tissue plasminogen activator? PMID- 17120252 TI - Effects of intraoperative hypothermia on neuropsychological outcomes after intracranial aneurysm surgery. AB - OBJECTIVE: Subarachnoid hemorrhage and surgical obliteration of ruptured intracranial aneurysms are frequently associated with neurological and neuropsychological abnormalities. We reported that intraoperative cooling did not improve neurological outcome in good-grade surgical subarachnoid hemorrhage patients, as assessed by the Glasgow Outcome Scale score or other neurological and functional measures (National Institutes of Health Stroke Scale, Rankin Disability Scale, Barthel Activities of Daily Living). We now report the results of neuropsychological testing in these patients. METHODS: A total of 1,001 patients who bled < or = 14 days before surgery were randomly assigned to intraoperative hypothermia (t = 33 degrees C) or normothermia (37 degrees C). Outcome was assessed approximately 3 months after surgery. Patients underwent the Benton Visual Retention, Controlled Oral Word Association, Rey-Osterrieth Complex Figure, Grooved Pegboard, and the Trail Making tests. T-scores for each test were calculated from normative data. T-scores were averaged to calculate a Composite Score. A test result (or the Composite Score) was considered "impaired" if the T score was two or more standard deviations below the norm. A Mini-Mental State Examination was also performed. RESULTS: Neurological outcome data were available in 1,000 patients. Sixty-one patients died. Of the 939 survivors, 873 completed 3 or more tests (exclusive of the Mini-Mental State Examination). Patients with poor neurological outcomes were less likely to complete testing; only 3.9% of Good Outcome (Glasgow Outcome Scale score = 1) patients were untested, compared with 38.6% of patients with Glasgow Outcome Scale scores of 3 and 4. There were no prerandomization demographic differences between the two treatment groups. For hypothermic patients, 16.8% were impaired from their Composite Score versus 20.0% of patients in the normothermic group (p = 0.317). For patients in the hypothermic group, 54.5% were impaired on at least one test, compared with 55.5% of patients in the normothermic group (p = 0.865). Similar results were seen in patients with baseline WFNS scores = I. Mini-Mental State Examination scores in the hypothermic and normothermic groups were 27.4 +/- 3.8 and 26.8 +/- 4.5, respectively. INTERPRETATION: This is the largest prospective evaluation of neuropsychological function after subarachnoid hemorrhage to date. Testing was completed in a high fraction of patients, demonstrating the feasibility of such testing in a large trial. However, the frequent inability to complete testing in poor-outcome patients suggests that testing may be best used to refine outcome assessments in good-grade patients. Many patients showed impairment on at least one test, with global impairment present in 17 to 20% of patients (18-21% of survivors). This was true even among the patients with the best preoperative condition (WFNS = 1). There was no difference in the incidence of impairment between hypothermic and normothermic groups. PMID- 17120255 TI - Recombinant therapeutics: from bench to bedside (if your health plan concurs). AB - A talented medical student at our institution was recently diagnosed with multiple sclerosis (MS). beta interferon was prescribed; however, her annual cap from student health insurance coverage for outpatient drug expenses is only Dollars 3000, a sum that would be exceeded within 3 months and leave her without coverage for symptomatic medications also required for her care. Unable to pay for the treatment from personal resources, she qualified to receive beta interferon without cost from a universal access program established by the manufacturer. Unfortunately, it was evident within a few months that she had a poor therapeutic response, so her physician prescribed natalizumab. Her health care plan requires that infusion therapy be given at the hospital's infusion center, yet for various reasons, the drug was not yet approved by the formulary. She remains unable to receive the drug. PMID- 17120258 TI - Measuring cognitive outcome after subarachnoid hemorrhage. PMID- 17120259 TI - Structure of a paramagnetic supermicroporous silica-based material via a multinuclear solid-state NMR monitoring. AB - The paramagnetic Si/Al material doped with 19.8% (weight) of Mn2+ has been characterized by the 1H, 13C, 27Al, 29Si MAS NMR spectra and the 1H, 29Si T1MAS relaxation measurements as a gel dried at 130 degrees C and as samples calcined at 504, 540 and 650 degrees C. It has been demonstrated that in spite of strong paramagnetic effects, NMR monitoring in combination with 29Si T1 relaxation experiments can be successful for structural descriptions of porous materials. PMID- 17120260 TI - Poly(methyl methacrylate) CE microchips replicated from poly(dimethylsiloxane) templates for the determination of cations. AB - A novel method for the rapid fabrication of poly(methyl methacrylate) (PMMA) microfluidic chips using poly(dimethylsiloxane) (PDMS) templates has been demonstrated. The PDMS molds were fabricated by soft lithography. The dense prepolymerized solution of methyl methacrylate containing thermal and UV initiators was allowed to polymerized between a PDMS template and a piece of a 1 mm thick commercial PMMA plate under a UV lamp. The images of microchannels on the PDMS template were precisely replicated into the synthesized PMMA substrates during the UV-initiated polymerization of the prepolymerized solution on the surface of the PMMA plate at room temperature. The polymerization could be completed within 10 min under ambient temperature. The chips were subsequently assembled by thermal bonding of the channel plate and the cover sheet. The new fabrication method obviates the need for specialized replication equipment and reduces the complexity of prototyping and manufacturing. Nearly 20 PMMA chips were replicated using a single PDMS mold. The attractive performance of the new microfluidic chips has been demonstrated by separating and detecting cations in connection with contactless conductivity detection. The fabricated PMMA microchip has also been successfully employed for the determination of potassium and sodium in environmental and biological samples. PMID- 17120261 TI - Usefulness of microchip electrophoresis for the analysis of mitochondrial DNA in forensic and ancient DNA studies. AB - We evaluate the usefulness of a commercially available microchip CE (MCE) device in different genetic identification studies performed with mitochondrial DNA (mtDNA) targets, including the haplotype analysis of HVR1 and HVR2 and the study of interspecies diversity of cytochrome b (Cyt b) and 16S ribosomal RNA (16S rRNA) mitochondrial genes in forensic and ancient DNA samples. The MCE commercial system tested in this study proved to be a fast and sensitive detection method of length heteroplasmy in cytosine stretches produced by 16 189T>C transitions in HVR1 and by 309.1 and 309.2 C-insertions in HVR2. Moreover, the quantitative analysis of PCR amplicons performed by LIF allowed normalizing the amplicon input in the sequencing reactions, improving the overall quality of sequence data. These quantitative data in combination with the quantification of genomic mtDNA by real-time PCR has been successfully used to evaluate the PCR efficiency and detection limit of full sequencing methods of different mtDNA targets. The quantification of amplicons also provided a method for the rapid evaluation of PCR efficiency of multiplex-PCR versus singleplex-PCR to amplify short HV1 amplicons (around 100 bp) from severely degraded ancient DNA samples. The combination of human-specific (Cyt b) and universal (16S rRNA) mtDNA primer sets in a single PCR reaction followed by MCE detection offers a very rapid and simple screening test to differentiate between human and nonhuman hair forensic samples. This method was also very efficient with degraded DNA templates from forensic hair and bone samples, because of its applicability to detect small amplicon sizes. Future possibilities of MCE in forensic DNA typing, including nuclear STRs and SNP profiling are suggested. PMID- 17120262 TI - The tautomeric forms of cyameluric acid derivatives. AB - The tautomerism of cyameluric acid C6N7O3H3 (1 a), cyamelurates and other heptazine derivatives has recently been studied by several theoretical investigations. In this experimental study we prepared stannyl and silyl derivatives of cyameluric acid (1 a): C6N7O3[Sn(C4H9)3]3 (3 a), C6N7O3[Sn(C2H5)3]3 (3 b), and C6N7O3[Si(CH3)3]3 (4). In order to investigate the structure of 1 a the mono- and dipotassium cyamelurate hydrates K(C6N7O3H2)2 H2O (5) and K2(C6N7O3H)1 H2O (6) were synthesized by UV/Vis-controlled titration of a potassium cyamelurate solution with aqueous hydrochloric acid. Compounds 3-6 were characterized by FTIR and solid-state NMR spectroscopy as well as simultaneous thermal analysis (TGA, DTA). The single crystal X-ray structures of the salts 5 and 6 show that the hydrogen atoms in both anions are localized on the peripheral nitrogen atoms. This indicates-in combination with the solid-state NMR studies that the most stable tautomer of solid 1 a is the triketo form with C3h symmetry. However, derivatives of both the hydroxyl and the amido tautomers may be formed depending on the substituent atoms: The spectroscopic data and single crystal structures of compounds C6N7O3[Si(CH3)3]3 (4) and the solvate C6N7O3[Sn(C2H5)3]3C2H4Cl2 (3 b') show that the former is derived from the symmetric trihydroxy form of 1 a, while 3 b' crystallizes as a chain-like polymer, which contains the tin atoms as multifunctional building blocks, that is, bridging pentacoordinated Et3SnO2 and Et3SnON units as well as non-bridging four-coordinated Et3SnN units. The cyameluric nucleus is part of the polymeric chains of C6N7O3[Sn(C2H5)3]3C2H4Cl2 (3 b'), by the action of both tautomeric forms of cyameluric acid, the amide and the ester form. PMID- 17120263 TI - Heavy-metal-ion capture, ion-exchange, and exceptional acid stability of the open framework chalcogenide (NH(4))(4)In(12)Se(20). AB - The hydrothermal synthesis of the purely inorganic open-framework indium selenide (NH(4))(4)In(12)Se(20) (1) is reported. Compound 1 exhibits a unique three dimensional open-framework structure. The framework of 1 shows an unusual, for a chalcogenide compound, rigidity arising from the unprecedented connection mode of its building blocks. Compound 1 possesses ion exchange capacity for Cs(+), Rb(+), NH(4) (+), but it has selectivity against Na(+) and Li(+). It also showed exceptional stability in relatively concentrated hydrochloric acid. Ion exchange of 1 with hydrochloric water solutions can produce its solid acid analogue H(2)(NH(4))(2)In(12)Se(20). The maximum cation-exchange capacity of 1 was found equal to two equivalents per mol, which is consistent with an exchange mechanism taking place in the 1D-channels formed by the largest cavities. In addition, 1 can do ion-exchange with heavy-metal ions like Hg(2+), Pb(2+), and Ag(+). The capacity of 1 to clean water solutions from heavy-metal ions was preliminarily investigated and found very high. Specifically, 1 can remove 99.9 % of Hg(2+), 99.8 % of Ag(+), and 94.9 % of Pb(2+) from aqueous solutions of each of these ions. Using different synthetic conditions, we isolated compound (NH(4))(2)In(12)Se(19) (2), which also has as good an acid stability as 1, but no ion-exchange properties. Overall, this work provides new hydrothermal synthetic approaches for isolation of all-inorganic open-framework chalcogenides. PMID- 17120264 TI - Chemical and constitutional influences in the self-assembly of functional supramolecular hydrogen-bonded nanoscopic fibres. AB - A new series of secondary amides bearing long alkyl chains with pi-electron-donor cores has been synthesized and characterised, and their self-assembly upon casting at surfaces has been studied. The different supramolecular assemblies of the materials have been visualized by using atomic force microscopy (AFM) and transmission electron microscopy (TEM). It is possible to obtain well-defined fibres of these aromatic core molecules as a result of the hydrogen bonds between the amide groups. Indeed, by altering the alkyl-chain lengths, constitutions, concentrations and solvent, it is possible to form different rodlike aggregates on graphite. Aggregate sizes with a lower limit of 6-8 nm width have been reached for different amide derivatives, while others show larger aggregates with rodlike morphologies which are several micrometers in length. For one compound that forms nanofibres, doping was performed by using a chemical oxidant, and the resulting layer on graphite was shown to exhibit metallic-like spectroscopy curves when probed with current-sensing AFM. This technique also revealed current maps of the surface of the molecular material. Fibre formation not only takes place on the graphite surface: nanometre scale rods have been imaged by using TEM on a grid after evaporation of solutions of the compounds in chloroform. Molecular modelling proves the importance of the hydrogen bonds in the generation of the fibres, and indicates that the constitution of the molecules is vital for the formation of the desired columnar stacks, results that are consistent with the images obtained by microscopic techniques. The results show the power of noncovalent bonds in self-assembly processes that can lead to electrically conducting nanoscale supramolecular wires. PMID- 17120265 TI - Rational design of sterically and electronically easily tunable chiral bisimidazolines and their applications in dual Lewis acid/bronsted base catalysis for highly enantioselective nitroaldol (Henry) reactions. AB - A new addition to the rational design of sterically and electrically easily tunable chiral bis(imidazoline) ligands from chiral amino alcohols has been developed. Vast structural variation of chiral bis(imidazoline) ligands can be simply achieved by the choice of both the 1,2-amino alcohol and its N-1 R1 substituent. A small library of chiral bisimidazolines (1 a-h) has been constructed. The method has provided an easy and simplified route to a diverse set of air-stable and water-tolerant chiral bis(imidazoline) ligands on 10 g scales. The dual Lewis Acid/Bronsted base catalytic system generated from the (S) 1 a/Cu(OTf)2 complex and Et3N was able to catalyze Henry reactions between aldehydes and nitromethane effectively at room temperature, and also to tolerate a wide scope of aldehydes with excellent enantiomeric excesses. Not only aromatic aldehydes but also aliphatic aldehydes afforded the nitroalcohol products, with enantiomeric excesses in the 93-98% range. This dual catalytic system is among the most effective systems so far reported for the asymmetric parent Henry reactions. This work also represents the first members of the class of chiral bisimidazolines to have been demonstrated to achieve excellent enantioselectivities. PMID- 17120266 TI - Configurations of nickel-cyclam antiviral complexes and protein recognition. AB - Nickel(II)-xylylbicyclam is a potent anti-HIV agent and binds strongly to the CXCR4 co-receptor. We have investigated configurational equilibria of Ni(II) cyclam derivatives, since these are important for receptor recognition. Crystallographic studies show that both trans and cis configurations are readily formed: [Ni(cyclam)(OAc)(2)] x H(2)O adopts the trans-III configuration with axial monodentate acetates, as does [Ni(benzylcyclam)(NO(3))(2)] with axial nitrate ligands, whereas [Ni(benzylcyclam)(OAc)](OAc)2 x H(2)O has an unusual folded cis-V configuration with Ni(II) coordination to bidentate acetate. UV/Vis and NMR studies show that the octahedral trans-III configuration slowly converts to square-planar trans-I in aqueous solution. For Ni(II)-xylylbicyclam, a mixture of cis-V and trans-I configurations was detected in solution. X-ray diffraction studies showed that crystals of lysozyme soaked in Ni(II)-cyclam or Ni(II) (2) xylylbicyclam contain two major binding sites, one involving Ni(II) coordination to Asp101 and hydrophobic interactions between the cyclam ring and Trp62 and Trp63, and the second hydrophobic interactions with Trp123. For Ni(II)-cyclam bound to Asp101, the cis-V configuration predominates. PMID- 17120267 TI - Diversity in guanine-selective DNA binding modes for an organometallic ruthenium arene complex. PMID- 17120268 TI - Design, synthesis and pharmacological evaluation of cyclic mimetics of the insulin-like peptide 3 (INSL3) B-chain. AB - Insulin-like peptide 3 (INSL3) is a peptide hormone belonging to the relaxin insulin superfamily of peptides that plays important roles in testes descent, oocyte maturation and the control of male germ cell apoptosis. These actions are mediated via a specific G-protein coupled receptor, LGR8. Previous structure activity studies have shown that the key binding site of INSL3 is situated within its B-chain. Recent studies in our laboratory have led to the identification of a cyclic peptide mimetic 2 of the INSL3 B-chain, which we have shown to compete with the binding of [33P]-relaxin to LGR8 expressed in HEK293T cells, and to inhibit cAMP-mediated signaling in these cells, i.e. it is an antagonist of INSL3. In order to further define the structure-activity relationships of cyclic analogues of the INSL3 B-chain, we used a structure-based approach to design a series of cyclic, disulfide-constrained INSL3 B-chain mimetics. To do this, we first created a model of the 3D structure of INSL3 using the crystal structure of human relaxin as a template. This model of INSL3 was then used as a template to design a series of disulfide-constrained mimetics of the INSL3 B-chain. The peptides were synthesized by solid-phase peptide synthesis using pseudoproline dipeptides to improve the synthesis outcome. Of the seven prepared INSL3 B-chain mimetics, three compounds were found to have partial displacement activity, while four were able to completely displace [33P]-relaxin from LGR8, including compounds that were markedly shorter than compound 2. The best of these, mimetic 6, showed significantly greater affinity for LGR8 than compound 2, but still displayed around 1000-fold less affinity for LGR8 than native INSL3. Analysis of selected mimetics for their alpha-helical content using circular dichroism (CD) spectroscopy revealed that, generally, the mimetics showed less than expected helicity. The inability of the compounds to display true native INSL3 structure is likely contributing to their reduced receptor binding affinity. We are currently examining alternative INSL3 B-chain mimetics that might better present key receptor binding residues in the native INSL3-like conformation. PMID- 17120269 TI - Top-down approach of completely sequencing a 4.9 kDa recombinant peptide using quadrupole time-of-flight mass spectrometry. AB - A recombinant peptide (near the C-terminal region of head involution defective protein) of 4.9 kDa has been completely sequenced and characterized using medium resolution mass spectrometry (QTOF). The observed difference in the experimental mass and the theoretical mass is due to beta-mercaptoethanol adduct formation on the cysteine residue. The fragmentation pattern is correlated with the primary structure of the protein. Top-down sequencing of the peptide was extended to small proteins like barstar of mass 10.3 kDa. PMID- 17120270 TI - Developing multiple high-throughput GLP methods for an investigational drug candidate in various matrices within a 96-well plate. AB - In early pharmaceutical product development, an investigational drug candidate is typically dosed to various species for toxicological and pharmacokinetic studies. Most of these studies require multiple analytical methods that have to be validated with good laboratory practice (GLP) prior to the application in regulated studies. Usually, these analytical methods are developed in either a serial or parallel approach. For either approach, the development of multiple analytical methods takes tremendous work from scientists and instruments, and thus is not cost-effective. In this respect, a new strategy has been developed for simultaneous GLP method development using liquid chromatographic separation and tandem mass spectrometric detection. This high-throughput approach allows system suitability, carryover, calibration curve, accuracy, precision, matrix effect and selectivity to be evaluated in one 96-well plate. The strategy has been successfully implemented for multiple investigational drug candidates at Abbott Laboratories. The methods developed with this strategy are accurate, precise, selective, robust and matrix-independent. As an example, ABT-279 was used to demonstrate the feasibility of this strategy. PMID- 17120271 TI - Liquid chromatography/tandem mass spectrometric study and analysis of xanthone and secoiridoid glycoside composition of Swertia chirata, a potent antidiabetic. AB - Swertia chirata is a bitter plant, used in the Indian system of medicine (Ayurveda) for various human ailments. The bioactive constituents include the xanthone and secoiridoid glycosides consisting of mangiferin, amarogentin, amaroswerin, sweroside and swertiamarin. Methanolic extracts of S. chirata possess constituents with antidiabetic activities, which was investigated by high performance liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS). Preliminary HPLC analyses were performed on a reversed-phase C18 column using gradient elution. In the LC/ESI-MS spectra, predominant [M+H]+ and [M+Na]+ ions were observed in positive ion mode and provided molecular mass information. The five components of S. chirata were structurally correlated and confirmed based on the fragmentation characteristics and information available in the literature. The fragmentation behavior of [M+H]+/[M+Na]+ ions of these components were deduced from the collision-induced dissociation (CID) spectra obtained from the selective on-column information dependant acquisition (IDA) approach. Xanthone-C-glycoside showed characteristic fragment ions due to fragmentation in the C-glycosidic unit while iridoid-O glycosides showed characteristic fragment ions due to cleavage in the glycoside linkage and retro-Diels-Alder (RDA) cleavage within an iridoid aglycone. Furthermore, on the basis of this information, an analytical assay was developed and validated to determine relative concentrations of mangiferin, amarogentin, amaroswerin, sweroside and swertiamarin. The detection was carried out using multiple reaction monitoring (MRM) in positive ionization mode with a total analysis time of 3.5 min. The method was successfully applied to standardize four different batches of herbal preparation on the basis of relative concentration of five bioactive components. PMID- 17120272 TI - 'On-the-fly' hydrogen/deuterium exchange liquid chromatography/mass spectrometry using a dual-sprayer atmospheric pressure ionisation source. AB - In-source 'on-the-fly' hydrogen/deuterium (H/D) exchange liquid chromatography mass spectrometry (LC/MS) has been investigated. The work was performed using a dual-sprayer source. The analyte was introduced through an electrospray ionisation sprayer and D2O was introduced through an atmospheric pressure chemical ionisation sprayer. To achieve H/D exchange sufficient to determine the number of exchangeable H atoms of a compound, a saturated 'steady-state' D2O atmosphere had to be created in the ion source by having a 2:1 or higher D2O-to analyte flow rate ratio. Under these conditions H/D exchange levels of 32-90% were achieved. In most cases the H/D exchange was sufficient to measure the number of exchangeable H atoms in some antiulcerative and anthelmintic pharmaceuticals. The concept of in-source 'on-the-fly' H/D exchange by introducing the deuterating agent via a second sprayer has been shown. It allows the integrity of the chromatographic separation to be kept, since the H/D exchange takes place post-separation. PMID- 17120273 TI - Bradykinin-related peptides from Phyllomedusa hypochondrialis azurea: Mass spectrometric structural characterisation and cloning of precursor cDNAs. AB - Amphibian skin secretions contain a plethora of bioactive compounds, many of which are understood to act to deter ingestion by predators. Bradykinins in particular are constitutively expressed in many amphibian skin secretions, mediating a variety of effects including hyperalgesia and contraction of gastric smooth muscle. Using a variety of proteomic techniques (high-performance liquid chromatography (HPLC) separation, matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS), and quadrupole time-of-flight tandem mass spectrometry (Q-TOF-MS/MS)) the current study identified 13 bradykinin-like peptides in the skin secretions of Phyllomedusa hypochondrialis azurea, including several new C-terminally extended isoforms (VPPGFTPFRLT, VHypPGFTPFRQT) and a novel phyllokinin-like peptide (RPPGFTPFRVY). Identification of the cDNA sequences encoding these peptides led to the deduction that the peptides were derived from differential post-translational processing and modification of five different precursors. Such an event emphasises the metabolic efficiency of peptide production in amphibian venom, with multiple products perhaps selective to different receptors in a variety of predators generated from a single precursor. An unusual modification was also recognised in the present study, with several bradykinin-like peptides featuring hydroxyprolination of the first proline residue rather than the commonly targeted second. This alteration may be mediated by the structural organisation of N-terminal amino acids prior to precursor processing. PMID- 17120274 TI - Characterization of ribosomal proteins as biomarkers for matrix-assisted laser desorption/ionization mass spectral identification of Lactobacillus plantarum. AB - For rapid identification of bacteria by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), a bioinformatics approach using ribosomal subunit proteins as biomarkers has been proposed. This method compares the observed masses for biomarkers with calculated masses as predicted from the amino acid sequences registered on protein databases. To evaluate this approach, the expressed ribosomal proteins of a genome-sequenced bacterium, Lactobacillus plantarum NCIMB 8826, were characterized as a model sample. The protein expression of 42 ribosomal subunit proteins, together with 10 ribosome associated proteins in the isolated ribosome fraction, was confirmed through two dimensional gel electrophoresis combined with peptide mass fingerprinting. The observed masses of the proteins in the isolated ribosome fraction were then determined by MALDI-MS. We preliminarily selected 44 biomarkers whose observed masses were matched with the calculated masses predicted from the amino acid sequence registered in the protein databases by considering N-terminal methionine loss only. Of these, the finally selected reliable biomarkers were 34 proteins including 31 ribosomal subunit proteins and 3 ribosome-associated proteins that could be observed in the MALDI mass spectra of the cell lysate sample. These biomarkers were usable in MALDI-MS characterization of two industrial L. plantarum cultures. PMID- 17120275 TI - Molecular weight determination of ultra-high mass compounds on a standard matrix assisted laser desorption/ionization time-of-flight mass spectrometer: PAMAM dendrimer generation 10 and immunoglobulin M. PMID- 17120276 TI - Mental health literacy in Japanese cancer patients: ability to recognize depression and preferences of treatments-comparison with Japanese lay public. AB - BACKGROUND: Insufficient knowledge about mental illness and its treatment has been shown to constitute a major barrier to its adequate care for mental illness in the lay public (LP). We therefore examined Japanese cancer patients' (CP) ability to recognize depression and their preferences of its treatments. PARTICIPANTS AND METHOD: One hundred lung CP and 300 LP were selected at random to participate in the study. Structured interviews using a vignette of a person with both cancer and depression were conducted with CP, and those using a vignette of a person with depression were carried out with LP, respectively. RESULTS: Only 11% of CP recognized the presence of depression in the vignette, while 25% of LP did (p<0.001). There were few significant differences in the preference for standard psychiatric treatments between CP and LP: standard treatments such as antidepressants (CP: 39%, LP: 36%) were less often rated as helpful, whereas non-standard treatments such as physical activity (CP: 85%, LP: 66%) were most often rated as helpful. CONCLUSIONS: The results indicated that cancer patients' knowledge about mental illness and its treatment were insufficient. Psychological education may reduce patient-related barriers to seek and to utilize optimal mental health care in cancer patients. PMID- 17120277 TI - The scent of urine spots of male mice, Mus musculus: Changes in chemical composition over time. AB - A dominant male mouse scent-marks his territory very frequently by emitting small urinary spots. The urine spots release in the air a variety of odorants that transmit different information to other mice, especially those concerning the time of deposition. To investigate this effect, small spots of urine of a dominant male mouse were left to freely release the odorants in the air for time intervals ranging from 0 min to 24 h prior to sampling. Thereupon, the odorants remaining in the spot were sampled at diffusion equilibrium (45 degrees C) in a small vial by means of headspace solid-phase microextraction followed by gas chromatography coupled to flame ionisation detection and mass spectrometry. Thirteen odorants were consistently found. Nine odorants were identified and four were matched. The rate of release of each odorant was characteristic and was described using principal component analysis. A first principal component was based on nine early odorants that showed a decreasing release over time. The odorants were 2,4-dehydro-exo-brevicomin, an unknown with 78% matching to 4 acetonilcycloheptanone, linalool, 2,4-dimethyl-phenol, 4-ethylphenol, indole, 2 butyl-1-octanol, an unknown with 83% matching to 2-ethyl-1-decanol, and 2,4-bis (1,1-dimethylethyl)phenol. A second principal component, based on two unknowns with 73% matching to yohimban-17-one and 71% matching to the 2-methyl-3-hydroxy 2,4,4-trimethyl ester of propanoic acid, had an irregular release after deposition. A third principal component of late odorants, based on 2-sec-butyl 4,5-dihydrothiazole and 6,10-dimethyl-5,9-undecaden-2-one, had a peak of release at about 22 min. In conclusion, the release of the odorants in the headspace of a urine spot may code and transmit information on the deposition time. PMID- 17120278 TI - The delta18O of root crown water best reflects source water delta18O in different types of herbaceous species. PMID- 17120279 TI - Origins and migratory routes of murine Cajal-Retzius cells. AB - The first layer that appears in the cortical neuroepithelium, the preplate, forms in the upper part of the cortex immediately below the pial surface. In mice, this layer exists between embryonic days (E) 10 and 13, and it hosts different cell populations. Here, we have studied the first cell population generated in the preplate, the Cajal-Retzius cells. There is considerable confusion regarding these cells with respect to both their site of generation and the migratory routes that they follow. This perhaps is due largely to the different opinions that exist regarding their characterization. We have studied the site of origin of these cells, their migratory routes, and the molecular markers that may distinguish them by injecting tracers into early embryos, culturing them in toto for 24 hours, and then performing immunohistochemistry. We found that the Cajal Retzius cells are most likely generated in the cortical hem by comparing with other cortical or extracortical origins. These cells are generated mainly at E10 and E11, and they subsequently migrate tangentially to cover the whole cortical mantle in 24 hours. From their site of origin in the medial wall of the telencephalon, they spread in a caudorostral direction, following an oblique migratory path toward the lateral part of the neuroepithelium. Prior to the splitting of the preplate, a percentage of the Cajal-Retzius cells that can be distinguished by the expression of reelin do not contain calretinin. Furthermore, there were no early-migrating neurons that expressed calbindin. PMID- 17120280 TI - Dynamics of bidirectional transport of Arc mRNA in neuronal dendrites. AB - The mRNA for Arc (activity-regulated cytoskeletal protein) is delivered into dendrites and localizes selectively at active synapses. Here we use a green fluorescent protein-based labeling system and confocal microscopy to define the transport kinetics of exogenously expressed mRNA from chimaeric Arc constructs (Arc/MS2 mRNA) in the dendrites of living rat neurons in culture. Arc/MS2 mRNA assembles into particles that move independently, bidirectionally, and intermittently in a fashion indicative of transport. Transport velocities range from below 6 to 65 mum/minute, which is consistent with actin-based and microtubule-based transport, respectively. In general, orthograde translocations are longer than retrograde translocations. Rapidly translocating Arc/MS2 mRNA particles sometimes reverse direction and decrease velocity just before stopping, suggesting that local signals regulate Arc mRNA targeting movements. These observations identify several phases of Arc mRNA movement that serve as potential points for regulating Arc mRNA localization. PMID- 17120281 TI - Origin of spectral tuning in rhodopsin--it is not the binding pocket. PMID- 17120282 TI - Manufacturing immunity to disease in a test tube: the magic bullet realized. AB - Although it took over one hundred years, Ehrlich's concept of the magic bullet is now a reality. Today, therapeutic antibodies are, arguably, the most important class of new drugs for the treatment of illnesses ranging from Alzheimer's disease to cancer. The emergence of therapeutic antibodies had to wait for advances in immunochemistry that allowed construction of antibodies in vitro. The centerpiece of the new technology is the combinatorial antibody library, which essentially allows one to synthesize an artificial immune system with a diversity that exceeds that of the natural repertoire. The construction of such libraries was perceived to be difficult because, if the natural immune system was to be used as the starting material, construction of the libraries would entail protocols that are the opposite of usual cloning. In gene cloning one starts with complexity and reduces it to a singularity. In the generation of diversity by construction of combinatorial antibody libraries, one starts with a collection of clones, randomly expands their complexity, and then returns them to recoverable singularities. The methods developed to accomplish this seemingly formidable task now allow construction of antibodies in a test tube to any antigen. These synthetic antibodies may be qualitatively and quantitatively superior to those of nature. PMID- 17120283 TI - A new catalytic route for the activation of sp-hybridized carbon-hydrogen bonds. PMID- 17120284 TI - Ultrahigh-resolution backbone structure of perdeuterated protein GB1 using residual dipolar couplings from two alignment media. PMID- 17120285 TI - Motoneurons of the flight power muscles of the blowfly Calliphora erythrocephala: structures and mutual dye coupling. AB - The morphologies of the motoneurons of the dorsolongitudinal and the three dorsoventral flight power muscles (DLM, DVM 1-3) of Calliphora were investigated by means of cobalt backfills and intracellular biocytin stainings. The DLM is innervated by four prothoracic motoneurons supplying the four ventral muscle fibers and one mesothoracic motoneuron supplying the two dorsal fibers. The three fibers of the DVM 1 and the two fibers of the DVM 2 are innervated by five mesothoracic motoneurons, whereas the two fibers of the DVM 3 are innervated by two prothoracic motoneurons. In general, the motoneurons of each muscle have a common ventral soma cluster located in a characteristic position on the ipsilateral side of the thoracic ganglion, show similar dendritic arborizations in the mesothoracic wing neuropil, and have the same axon pathway. Only the soma of the common motoneuron of two dorsal fibers of the DLM is situated dorsally in the contralateral hemiganglion. The motoneurons of each muscle were found to be strongly dye coupled with each other, indicating that they are connected by gap junctions. In addition, the motoneurons of each muscle establish characteristic coupling patterns with the motoneurons of the other flight power muscles on both sides of the thorax and with two bilateral groups of local mesothoracic interneurons. The revealed coupling patterns are assumed to be of major relevance for the generation the characteristic, rhythmic flight activity of the motoneurons described in previous studies. PMID- 17120286 TI - Kappa opioid receptors in the rostral ventromedial medulla of male and female rats. AB - Kappa opioid receptor (KOR) ligands alter nociceptive responses when applied to the rostral ventromedial medulla (RVM). However, the effects of kappa opioid receptor ligands are distinct in males and females. The present study examined the distribution of kappa opioid receptor immunoreactivity in the RVM of male and female rats. KOR immunoreactivity was found at pre- and postsynaptic sites within the RVM of both sexes. The most common KOR-immunoreactive (KOR-ir) neuronal structures were unmyelinated axons, followed by axon terminals, dendrites, and somata. Different proportions of KOR-ir axon terminals and dendrites were found in females at different estrous stages. Specifically, dendrites containing KOR immunoreactivity were less abundant in proestrus females compared with estrus females and showed a trend toward being less abundant in males, suggesting that KOR ligands applied to the RVM may be less potent in proestrus females. These findings suggest that the distribution of KORs in the RVM may be influenced by reproductive hormone levels. We also found KOR immunoreactivity in many spinally projecting neurons within the RVM of female rats. These findings are consistent with the hypothesis that KOR ligands influence nociceptive behaviors by altering the activity of specific populations of neurons within the RVM. The abundance of KOR in axons and axon terminals in RVM indicates a substantial role for presynaptic effects of KOR ligands through pathways that have not been clearly delineated. Altering the balance between pre- and postsynaptic receptive sites may underlie differences in the effects of KOR agonists on nociceptive responses in males and females. PMID- 17120287 TI - Enriched expression and developmental regulation of the middle-weight neurofilament (NF-M) gene in song control nuclei of the zebra finch. AB - Songbirds evolved a complex set of dimorphic telencephalic nuclei that are essential for the learning and production of song. These nuclei, which together make up the oscine song control system, present several neurochemical properties that distinguish them from the rest of the telencephalon. Here we show that the expression of the gene encoding the middle-weight neurofilament (NF-M), an important component of the neuronal cytoskeleton and a useful tool for studying the cytarchitectonic organization of mammalian cortical areas, is highly enriched in large neurons within pallial song control nuclei (nucleus HVC, robustus nucleus of the arcopallium, and lateral magnocellular nucleus of the nidopallium) of male zebra finches (Taeniopygia guttata). We also show that this transcript is highly expressed in large neurons in the medulla, pons, midbrain, and thalamus. Moreover, we demonstrate that NF-M expression in song control nuclei changes during postembryonic development, peaking during an early phase of the song learning period that coincides with the maturation of the song system. We did not observe changes in NF-M expression in auditory areas or in song control nuclei in the contexts of hearing song or singing, although these contexts result in marked induction of the transcription factor ZENK. This observation suggests that NF-M might not be under the regulatory control of ZENK in auditory areas or in song control nuclei. Overall, our data indicate that NF-M is a neurochemical marker for pallial song control nuclei and provide suggestive evidence of an involvement of NF-M in the development and/or maturation of the oscine song control system. PMID- 17120288 TI - Peptidomics of identified neurons demonstrates a highly differentiated expression pattern of FXPRLamides in the neuroendocrine system of an insect. AB - FXPRLamides are insect neuropeptides that mediate such diverse functions as pheromone biosynthesis, visceral muscle contraction, and induction of diapause. Although multiple forms occur in every insect studied so far, little is known about a possible functional differentiation and/or differences in the cellular expression pattern of these messenger molecules. In this study, we performed a mass spectrometric survey of all FXPRLamide-expressing neurosecretory neurons in the CNS of Periplaneta americana. That species combines a very well characterized peptidergic system with relatively easy accessible neurosecretory cells suitable for dissection. In addition to the extensive mass spectrometric analyses of single cells, the projection of the FXPRLamide-expressing neurons was studied with three antisera specifically recognizing different FXPRLamides. The following conclusions can be drawn from this first comprehensive peptidomic approach on insect neurons. 1) A high degree of differentiation in the expression of FXPRLamides exists; not fewer then four cell types containing different sets of FXPRLamides were observed. 2) A low level of colocalization with other neuropeptides was found in these neurons. 3) A comparison with FXPRLamide expressing neurons of other insects shows a high degree of conservation in the localization and projection of these neurons, which is not corroborated by a similar conservation of the corresponding peptide sequences. 4) Although the methods for cell identification, dissection, and sample preparation for mass spectrometry were kept as simple as possible, it was unambiguously shown that this approach is generally suitable for routine analysis of single identified neurons of insects. PMID- 17120289 TI - Postsynaptic targets of somatostatin-containing interneurons in the rat basolateral amygdala. AB - The basolateral amygdala contains several subpopulations of inhibitory interneurons that can be distinguished on the basis of their content of calcium binding proteins or peptides. Although previous studies have shown that interneuronal subpopulations containing parvalbumin (PV) or vasoactive intestinal peptide (VIP) innervate distinct postsynaptic domains of pyramidal cells as well as other interneurons, very little is known about the synaptic outputs of the interneuronal subpopulation that expresses somatostatin (SOM). The present study utilized dual-labeling immunocytochemical techniques at the light and electron microscopic levels to analyze the innervation of pyramidal cells, PV+ interneurons, and VIP+ interneurons in the anterior basolateral amygdalar nucleus (BLa) by SOM+ axon terminals. Pyramidal cell somata and dendrites were selectively labeled with antibodies to calcium/calmodulin-dependent protein kinase II (CaMK); previous studies have shown that the vast majority of dendritic spines, whether CAMK+ or not, arise from pyramidal cells. Almost all SOM+ axon terminals formed symmetrical synapses. The main postsynaptic targets of SOM+ terminals were small-caliber CaMK+ dendrites and dendritic spines, some of which were CaMK+. These SOM+ synapses with dendrites were often in close proximity to asymmetrical (excitatory) synapses to these same structures formed by unlabeled terminals. Few SOM+ terminals formed synapses with CaMK+ pyramidal cell somata or large-caliber (proximal) dendrites. Likewise, only 15% of SOM+ terminals formed synapses with PV+, VIP+, or SOM+ interneurons. These findings suggest that inhibitory inputs from SOM+ interneurons may interact with excitatory inputs to pyramidal cell distal dendrites in the BLa. These interactions might affect synaptic plasticity related to emotional learning. PMID- 17120290 TI - Organization of endogenous opioids in the rostral agranular insular cortex of the rat. AB - The rostral agranular insular cortex (RAIC) of rats has opioid receptors and has been implicated in the analgesic and reinforcing effects of opiates. To help in understanding the function of endogenous opioids in this structure, we sought to identify and describe the opioid peptides intrinsic to the RAIC by using immunohistochemical methods. Immunolabeling for proopiomelanocortin (POMC), the precursor to beta-endorphin, and endomorphin 1 and 2 on sectioned rat forebrain revealed limited labeling consisting of individual varicose fibers. Immunolabeling for prodynorphin and enkephalin revealed numerous immunopositive cell bodies and fibers with distribution and morphology unique to each. Prodynorphin-immunopositive cell bodies consisted of two types: large, lightly labeled, pyramidal-shaped cell bodies in lamina V and more intensely labeled, small, ovoid cell bodies scattered in other lamina. Axonal fibers immunolabeled for prodynorphin varied in size and were found in all lamina. Immunolabeling for the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) was rarely found in dynorphin-containing cell bodies (6%, 10/167) but was visible within a subpopulation of axons. Enkephalin immunolabeling was detected within a single morphological subpopulation of nonpyramidal neurons located predominantly in lamina II/III, 30% (33/109) of which were also GABA immunopositive. Axons immunolabeled for enkephalin were also abundant in lamina II/III. These results suggest that dynorphin and enkephalin peptides are the predominant endogenous opioids in the RAIC and their distinct distributions suggest divergent functional roles. The localization of prodynorphin immunoreactivity to pyramidal cells suggests the possibility that this neuropeptide may be used in RAIC projection neurons, whereas enkephalin distribution was more characteristic of a role in local networks. PMID- 17120291 TI - Differential mRNA distribution of components of the ERK/MAPK signalling cascade in the adult mouse brain. AB - The mitogen-activated protein kinases (MAPKs), also called extracellular signal regulated kinases (ERKs), are a group of serine/threonine terminal protein kinases activated downstream of a pleiotrophy of transmembrane receptors. Main intracellular components of the MAPK signalling pathway are the RAF, MEK, and ERK proteins, which work in a cascade of activator and effector proteins. They regulate many fundamental cellular functions, including cell proliferation, cell survival, and cell differentiation by transducing extracellular signals to cytoplasmic and nuclear effectors. To reveal more details about possible activation cascades in this pathway, the present study gives a complete description of the differential expression of Braf, Mek1, Mek2, Mek5, Erk1, Erk2, Erk3, and Erk5 in the adult murine brain by way of in situ hybridization analysis. In this study, we found that each gene is widely expressed in the whole brain, except for Mek2, but each displays a very distinct expression pattern, leading to distinct interactions of the MAPK components within different regions. Most notably we found that 1) Braf and Erk3 are coexpressed in the hippocampus proper, confirming a possible functional interaction; 2) in most forebrain areas, Mek5 and Erk5 are coexpressed; and 3) in the neurogenic regions of the brain, namely, the olfactory bulb and the dentate gyrus, Braf is absent, indicating that other activator proteins have to take over its function. Despite these differences, our results show widespread coexpression of the pathway components, thereby confirming the hypothesis of redundant functions among several MEK and ERK proteins in some regions of the brain. PMID- 17120292 TI - Region-specific expression and sex-steroidal regulation on aromatase and its mRNA in the male rat brain: immunohistochemical and in situ hybridization analyses. AB - The brain has an estrogen-biosynthetic potential resulting from the presence of neuronal aromatase, which controls the intraneural sex-steroidal milieu and is involved in brain sexual differentiation, psychobehavioral regulation, and neuroprotection. In the rat brain, three distinct aromatase-P450-immunoreactive (AromP450-I) neural groups have been categorized in terms of their peak expression time (fetal, fetoneonatal, and young-to-adult groups), suggesting the presence of region-specific regulation on brain AromP450. In the present study, we compared the expressions between AromP450 protein and mRNA by using immunohistochemistry and in situ hybridization with an ovary-derived cRNA probe in serial sections of fetal, fetoneonatal, and adult male rat brains and then performed steroidal manipulations to evaluate the sex-steroidal effects on AromP450 in adult orchiectomized and adrenalectomized (OCX + ADX) male rats. As a result, prominent mRNA signals were detected in the fetal (i.e., the anterior medial preoptic nucleus) and fetoneonatal (i.e., the medial preopticoamygdaloid neuronal arc) groups, although no detectable signal was found in the "young-to adult" group (i.e., the central amygdaloid nucleus). In addition, the "fetoneonatal" AromP450-I neurons were prominently reduced in number and intensity after OCX + ADX and then were reinstated by the administration of dihydrotestosterone, testosterone, or 17beta-estradiol. In contrast, none of the sex steroids had any significant effects on the young-to-adult group. Several possible explanations were explored for why the young-to-adult group may differ in aromatase expression and regulation, including the possibility that distinct splicing variants or isozymes for aromatase exist in the rat brain. PMID- 17120293 TI - Adult neurogenesis: a common strategy across diverse species. AB - Adult neurogenesis, the generation of new neurons from adult precursor cells, occurs in the brains of a phylogenetically diverse array of animals. In the higher (amniotic) vertebrates, these precursor cells are glial cells that reside within specialized regions, known as neurogenic niches, the elements of which both support and regulate neurogenesis. The in vivo identity and location of the precursor cells responsible for adult neurogenesis in nonvertebrate taxa, however, remain largely unknown. Among the invertebrates, adult neurogenesis has been particularly well characterized in freshwater crayfish (Arthropoda, Crustacea), although the identity of the precursor cells sustaining continuous neuronal proliferation in these animals has yet to be established. Here we provide evidence suggesting that, as in the higher vertebrates, the precursor cells maintaining adult neurogenesis in the crayfish Procambarus clarkii are glial cells. These precursor cells reside within a specialized region, or niche, on the ventral surface of the brain, and their progeny migrate from this niche along glial fibers and then proliferate to form new neurons in the central olfactory pathway. The niche in which these precursor cells reside has many features in common with the neurogenic niches of higher vertebrates. These commonalities include: glial cells functioning as both precursor and support cells, directed migration, close association with the brain vasculature, and specialized basal laminae. The cellular machinery maintaining adult neurogenesis appears, therefore, to be shared by widely disparate taxa. These extensive structural and functional parallels suggest a common strategy for the generation of new neurons in adult brains. PMID- 17120295 TI - In vivo analysis of the post-natal development of normal mouse brain by DTI. AB - The water diffusion characteristics of wild-type mouse brains have been studied in vivo by DTI to follow developmental changes. Here, axial (lambda(//)) and radial (lambda(perpendicular)) diffusivities and fractional anisotropy were measured from the fifth day of life (P5) and at three other post-natal ages (P12, P19 and P54). Magnetic resonance images were collected from a single sagittal slice in the middle of the two hemispheres; ROI were chosen in nine different structures of both grey and white matter. Fractional anisotropy (FA) from P5 onwards distinguished structures of both white and grey matter, even though myelination had yet to occur. Between P5 and P54, a significant increase in FA was observed in the genu of the corpus callosum due to a significant decrease in lambda(perpendicular) whereas lambda(//) remained stable. Many other significant variations of lambda(//) and lambda(perpendicular) were measured in different structures. They were substantially correlated with axon and myelin maturation which are responsible for the main evolutions of the brain during its post-natal development. These quantitative data show that in vivo characterization of the anatomy and microstructure of the normal mouse brain during development is possible. The normative data will greatly improve the characterization of abnormal development in the transgenic mouse brain. PMID- 17120296 TI - Comparison between prospective and retrospective triggering for mouse cardiac MRI. AB - High-resolution magnetic resonance imaging (MRI) has evolved into one of the major non-invasive tools to study the healthy and diseased mouse heart. This study presents a Cartesian CINE MRI protocol based on a fast low-angle shot sequence with a navigator echo to generate cardiac triggering and respiratory gating signals retrospectively, making the use of ECG leads and respiratory motion sensors obsolete. MRI of the in vivo mouse heart using this sequence resulted in CINE images with no detectable cardiac and respiratory motion artefacts. The retrospective method allows for steady-state imaging of the mouse heart, which is essential for quantitative contrast-enhanced MRI studies. A comparison was made between prospective and retrospective methods in terms of the signal-to-noise ratio and the contrast-to-noise ratio between blood and myocardial wall, as well as global cardiac functional indices: end-diastolic volume, end-systolic volume, stroke volume and ejection fraction. The retrospective method resulted in almost constant left-ventricle wall signal intensity throughout the cardiac cycle, at the expense of a decrease in the signal-to-noise ratio and the contrast-to-noise ratio between blood and myocardial wall as compared with the prospective method. Prospective and retrospective sequences yielded comparable global cardiac functional indices. The largest mean relative difference found was 8% for the end-systolic volume. PMID- 17120294 TI - Selective enrichment of DJ-1 protein in primate striatal neuronal processes: implications for Parkinson's disease. AB - Mutations in DJ-1 cause autosomal recessive, early-onset Parkinson's disease (PD). The precise function and distribution of DJ-1 in the central nervous system remain unclear. In this study, we performed a comprehensive analysis of DJ-1 expression in human, monkey, and rat brains with antibodies that recognize distinct, evolutionarily conserved epitopes of DJ-1. We found that DJ-1 displays region-specific neuronal and glial labeling in human and nonhuman primate brain, sharply contrasting with the primarily neuronal expression pattern observed throughout rat brain. Further immunohistochemical analysis of DJ-1 expression in human and nonhuman primate brains showed that DJ-1 protein is expressed in neurons within the substantia nigra pars compacta and striatum, two regions critically involved in PD pathogenesis. Moreover, immunoelectron microscopic analysis revealed a selective enrichment of DJ-1 within primate striatal axons, presynaptic terminals, and dendritic spines with respect to the DJ-1 expression in prefrontal cortex. Together, these findings indicate neuronal and synaptic expression of DJ-1 in primate subcortical brain regions and suggest a physiological role for DJ-1 in the survival and/or function of nigral-striatal neurons. PMID- 17120297 TI - Examination of the known-groups validity of the Liebowitz Social Anxiety Scale. AB - We examined the known-groups validity of the Liebowitz Social Anxiety Scale (LSAS) by comparing the scores of patients with social anxiety disorder (n=46), generalized anxiety disorder plus an additional diagnosis of social anxiety disorder (n=15), generalized anxiety disorder without social anxiety disorder (n=12), and nonanxious controls (n=34). The LSAS total score discriminated significantly among all pairs of groups. Similar analyses were conducted on the original LSAS subscales and additional subscales derived from the factor-analytic work of Safren et al. [1999]. Original subscales showed a pattern substantially similar to that of the total score, but subscale intercorrelations and total score-subscale correlations were extremely high, suggesting that these subscales do not provide much unique information beyond that provided by the total score. Factor-analytically derived subscales were less highly correlated with each other or with the LSAS total score. Although the pattern of differences was more variable across subscales, the factor-analytically derived subscales, in conjunction with the total score, may provide more nonredundant information of clinical relevance than the original subscales. Limitations and future directions for research on the LSAS are discussed. PMID- 17120298 TI - Electrospray ionization mass spectrometry fingerprinting of perfumes: Rapid classification and counterfeit detection. AB - A fast procedure to classify perfumes and identify counterfeit samples is described. Dilution of a few microL of the sample in a 1:1 methanol/water solution is followed by detection of its major polar components via direct infusion electrospray ionization mass spectrometry (ESI-MS) in the positive ion mode. As proof-of-principle cases, three famous brands of perfumes were used. The ESI+-MS fingerprints of authentic samples were very characteristic, showing distinctive sets of polar markers for each sample. Principal component analysis (PCA) placed samples of the three perfume brands in well-defined groups. Counterfeit samples were also clearly detected owing to contrasting ESI-MS fingerprints, with PCA placing these samples far away from the authentic samples. PMID- 17120299 TI - Simultaneous determination of seven active components of Fufang Danshen tablet by high performance liquid chromatography. AB - A high-performance liquid chromatography method was established for simultaneously determining seven major components, i.e. protocatechuic aldehyde, notoginsenoside R(1), ginsenoside Rg(1), salvianolic acid B, ginsenoside Rb(1), cryptotanshinone and tanshinone IIA in Fufang Danshen tablet, a commonly used traditional Chinese medicinal combined prescription mainly derived from the roots of Salvia miltiorrhiza and Panax notoginseng. These seven compounds, belonging to the chemical types of phenolic acids, diterpenoid quinones and saponins, were simultaneously separated on Zorbax C(18) column (250 x 4.6 mm, 5.0 microm) with the column temperature at 30 degrees C. The mobile phase was composed of (A) aqueous phosphoric acid (0.1%, v/v) and (B) acetonitrile using a gradient elution of 7-17% B at 0-10 min, 17-20% B at 10-12 min, 20-21% B at 12-16 min, 21% B at 16 32 min, 21-29% B at 32-40 min, 29-35% B at 40-55 min, 35-65% B at 55-65 min and 65-80% B at 65-80 min; the flow rate was 1.0 mL/min. Detection wavelengths were set at 203 nm for notoginsenoside R(1), ginsenoside Rg(1) and ginsenoside Rb(1), 281 nm for protocatechuic aldehyde, salvianolic acid B, and 270 nm for cryptotanshinone and tanshinone IIA. All calibration curves showed good linear regression (r(2) > 0.9992) within test ranges. The established method showed good precision and accuracy with overall intra-day and inter-day variations of 0.15 4.35 and 0.61-5.17% respectively, and overall recoveries of 94.8-102.1% for the seven compounds analyzed. The developed method has been successfully applied to simultaneous evaluation of the intrinsic quality of both Danshen and Sanqi in Fufang Danshen tablets from different pharmaceutical companies. PMID- 17120300 TI - Investigation of lipophilicity of anticancer-active thioquinoline derivatives. AB - The lipophilicity of a series of anticancer propargylthioquinoline derivatives has been investigated using chromatographic and computational methods. The parameters of relative lipophilicity (R(MO) and logk0) of the tested compounds were determined experimentally both by reversed-phase thin layer (RP-TLC), and high-performance liquid chromatographic methods (RP-HPLC, LiChrospher RP18 column), with mixtures of acetonitrile and water as mobile phases. Their phospholipophilicity (logk(0IAM)) was determined using immobilized artificial membrane HPLC (IAM. PC DD2 Regis column). Mobile phase acetonitrile concentrations were in the ranges 50-90% (RP-TLC), 55-90% (RP-HPLC) and 35-60% (IAM-HPLC). The R(M), logk and logk(IAM) values of the compounds investigated were linearly dependent on acetonitrile concentration. The analysis led to the calculation of R(MO), logk0 and logk(0IAM) parameter values for each of the tested compounds. Their partition coefficients (logP) were also calculated with the Pallas and CAChe programs. The obtained results indicated that, among experimental methods, both RP-TLC and RP-HPLC gave similar results, and these methods enable the determination of lipophilicity of derivatives of thioquinolines. Using the IAM-HPLC technique a simple method of estimation of phospholipoplilicity was described. The CAChe program might better predict calculated lipophilicity logP values, and therefore is a useful tool for the early stage of design of new propargyl thioquinolines. PMID- 17120301 TI - A liquid chromatography-mass spectrometry method for the quantitation of aristololactam-I in rat plasma. AB - A sensitive method with liquid chromatography-electrospray ionization mass spectrometry has been developed and validated for the determination of aristololactam-I in rat plasma after oral administration of aristolochic acid-I using finesteride as the internal standard. Chromatographic separation was achieved on a Lichrospher C(18) column using methanol:0.05% acetic acid in water (71:29, v/v) as a mobile phase delivered at a flow rate of 1 mL/min. The assay was linear for aristololactam-I over the range 0.3-300 ng/mL. The analysis of quality control samples demonstrated precision with coefficient of variation less than 20% (n = 5). Absolute recovery of aristololactam-I was 90.4-97.3%. The LC-MS method for the determination of aristololactam-I is sensitive, specific and can be used to investigate the toxicokinetics of aristololactam-I. PMID- 17120302 TI - Pesticide multiresidue analysis in Panax ginseng (C. A. Meyer) by solid-phase extraction and gas chromatography with electron capture and nitrogen-phosphorus detection. AB - An analytical multi-residue method using gas chromatography coupled with electron capture and a nitrogen-phosphorus detector was investigated for the simultaneous determination of 18 commonly used insecticides and fungicides in Korean ginseng (Panax ginseng C. A. Meyer). Samples were previously extracted with an acetonitrile and cleaned up by solid-phase extraction (SPE). The calibration curves were linear, with determination coefficients higher than 0.989. Recoveries at concentrations between 0.01 and 14.9 ppm ranged from 72.3 to 117.2%, with precision, which was expressed as relative standard deviation (RSD), at values lower than 5%. The proposed method was applied to the determination of pesticide levels from 12 ginseng samples, taken from four different agricultural areas of Jeonnam province, where several insecticides and fungicides were applied. Except in one sample, tolclofos-m was the only pesticide contained at a level lower than the maximum residue limits (MRL) authorized by the Korea Food and Drug Administration (KFDA) in real ginseng samples grown for 4, 5 and 6 years. PMID- 17120303 TI - High-performance liquid chromatography and LC-ESI-MS method for the identification and quantification of two biologically active polyisoprenylated benzophenones xanthochymol and isoxanthochymol in different parts of Garcinia indica. AB - A reverse-phase high-performance liquid chromatographic method was developed for qualitative and quantitative analysis of xanthochymol (1), and isoxanthochymol (2) in the fruit rinds, leaves and seed pericarps of Garcinia indica with confirmation using PDA detection and electrospray ionization MS. Absorption at 276 nm was chosen as the measuring wavelength at which resolution and baseline separation of compounds (1) and (2) could be obtained. The identity of the above two isomeric compounds (1) and (2) in the samples was unambiguously determined by their respective quasi-molecular ion [M - H]- in ESI-MS. Compounds (1) and (2) were qualitatively and quantitatively analyzed in the above three samples of Garcinia indica. The overall analytical procedure is rapid and reproducible and is considered for the analysis of the above two compounds. PMID- 17120304 TI - Post TLC developing technique for tyrosinase inhibitor detection. AB - Post TLC developing technique was developed to detect substances which can inhibit tyrosinase activity. The method involved spraying the TLC plate or chromatographic paper containing sample spot(s) with tyrosinase and l-tyrosine solutions successively. A positive result could be visualized directly as white spot(s) against a brownish-purple background. The method can either be used as a quick screening method for tyrosinase inhibitor detection or a guiding procedure for an isolation of tyrosinase inhibitors from mixtures or natural product extracts. The technique is sensitive enough to give a clear result in the presence of only 6 ng glabridin. PMID- 17120305 TI - Pharmacokinetic study of free-form sinomenine in rat skin by microdialysis coupled with liquid chromatography-electrospray mass spectrometry. AB - Sinomenine (7,8-didehydro-4-hydroxy-3,7-dimethoxy-17-methylmorphinan-6-one) is a pure alkaloid extracted from the Chinese medical plant. In this report a liquid chromatography-electrospray mass spectrometry (LC-ESI-MS) method with in vivo microdialysis for the pharmacokinetic study of free-form sinomenine in rat skin has been developed. A microdialysis probe was surgically implanted into the subcutaneous tissue of the rats and an isotonic phosphate buffer (PBS) was used as the perfusion medium. Samples were collected and then analyzed off-line by LC ESI-MS. The chromatographic separation was achieved within 4.2 min by using a narrow-bore Xterra C(18) column (2.1 x 150 mm, 5 microm) with acetonitrile-(10 mmol/L ammonium acetate buffer, 0.1% acetic acid) (15:85, v/v). Ion signal m/z 330.1 for sinomenine was measured in the positive mode. Linearity was established for the range of concentrations of 2.0-10000.0 ng/mL with a coefficient of determination (r) of 0.9989. The intra- and inter-day reproducibility of the present method was better than 6%. The lower limit of quantification (LLOQ) was 1.0 ng/mL. The proposed method described provides more authentic information on pharmacokinetics and metabolism at the site of action by using the coupling of microdialysis to LC-ESI-MS technique than the traditional sampling methods. PMID- 17120306 TI - Influence of in vitro oxygen concentrations on preimplantation embryo development, gene expression and production of Hanwoo calves following embryo transfer. AB - This study evaluated the effects of two different oxygen (O2) concentrations on in vitro embryo development, embryo quality, and gene expression and the in vivo development following embryos transfer to recipients of natural and synchronized estrus in bovines. Cumulus oocyte complexes were in vitro matured in TCM199 supplemented with FSH (10 microg/ml), LH (10 microg/ml), and 10% (v/v) FBS. Presumptive zygotes were cultured in SOF medium either under 5% (low) or 20% (high) O2 in air. Cleavage rates did not differ between groups. Blastocyst and hatched blastocyst development in 5% O2 were significantly (P < 0.05) higher than in 20% O2. Total cell number of in vivo blastocyst was significantly (P < 0.05) higher than that of in vitro blastocyst. ICM ratio and apoptosis of in vivo blastocyst were significantly (P < 0.05) lower than that of in vitro blastocyst. Using real time PCR, we have found that for the set of genes (GLUT-1, MnSOD, VEGF, Bax, and Bcl-2) analyzed, there were differences in mRNA expression between in vitro produced (IVP) and in vivo produced embryos. Interestingly, the abundance of transcript for IFN-tau in IVP embryos produced under 5% O2 concentration was similar to in vivo counterparts. The pregnancy and twin rates of natural recipients were significantly (P < 0.05) higher than those of synchronized counterparts. No significant difference in the offspring sex was observed. In conclusion, low (5%) O2 concentration during IVC was beneficial for enhancing the embryo quality and recipients of natural estrus were more suitable than synchronized estrus for stable production of Hanwoo calves. PMID- 17120307 TI - Expression of vascular endothelial growth factor and its receptors in the porcine corpus luteum during the estrous cycle and early pregnancy. AB - The present study was undertaken to determine the expression of vascular endothelial growth factor (VEGF) and its receptors, fms-like tyrosine kinase (Flt 1) and fetal liver kinase-1/kinase insert domain-containing receptor (Flk-1/KDR), in the porcine corpus luteum (CL) during the estrous cycle and early pregnancy. Immunohistochemical studies localized proteins of VEGF ligand-receptor system in the cytoplasm of luteal cells and in some blood vessels. Western blot analysis revealed significantly higher levels of VEGF protein during early and mid-luteal phase (vs. late luteal phase; P<0.001 and P<0.01, respectively). Quantification of VEGF mRNA in the CL showed increased mRNA levels during entire luteal phase (vs. Days 16-17; P<0.05). Expression of Flt-1 protein remained high during luteal phase (P<0.001), but the mRNA levels tended to increase from the early to the late luteal phase. Elevated protein expression of Flk-1/KDR was found in the mid luteal phase (vs. Days 16-17; P<0.05). However, induction of Flk-1/KDR mRNA expression occurred earlier, in early luteal phase. The lowest VEGF, Flt-1 and Flk-1/KDR mRNA and protein levels were observed in regressed CL (P<0.001). During pregnancy, VEGF, Flt-1 and Flk-1/KDR mRNA and protein expression was comparable to the mid-luteal phase. In conclusion, the present study has demonstrated dynamic expression of VEGF and its receptors in the porcine CL during the estrous cycle and early pregnancy. These data suggest that the VEGF ligand-receptor system may play an important role in the development and maintenance of the CL in pigs. PMID- 17120308 TI - In utero exposure to tributyltin alters the expression of E-cadherin and localization of claudin-1 in intercellular junctions of the rat ventral prostate. AB - Tributyltin (TBT) is an environmental contaminant, exhibiting well-established toxicity to reproductive systems in aquatic organisms. Little information exists regarding the effects of TBT on mammalian reproduction. Cellular junctions are crucial for sperm development and maturation. Intercellular tight junctions are formed by transmembrane proteins such as claudins (Cldns), while the formation of tight junctions involves signaling components of adhering junctions, comprised of cadherins. The objectives of this study were to determine the effects of in utero exposure to TBT on the rat ventral prostate. Pregnant Sprague-Dawley rats were given doses of TBT (2.5, 10, or 20 mg/kg) throughout gestation and sacrificed at Day 91. Ventral prostate weights of TBT-treated rats were decreased in all treatment groups. Results of gene expression macro-array analysis indicated that numerous genes related to cellular adhesion and cell polarity were affected. Cldn 1 mRNA levels decreased after exposure to TBT. Cldn-1 was immunolocalized to the apical lateral margins of adjacent prostatic epithelial cells in controls, but was increasingly dispersed along the lateral plasma membrane with increasing TBT dose, suggesting that the targeting of Cldn-1 or its localization to tight junctions was altered as a result of fetal TBT exposure. E-cadherin mRNA levels and immunolocalization were decreased in a dose-dependent manner. These data indicate that in utero TBT exposure results in permanent alterations in ventral prostate and that these are associated with alterations in the expression and distribution of cell adhesion and tight junctional proteins. PMID- 17120309 TI - Comprehensive genomic analysis identifies MDM2 and AURKA as novel amplified genes in juvenile angiofibromas. AB - BACKGROUND: Frequent beta-catenin mutations have been detected in juvenile angiofibromas, but the tumor pathogenesis remains unknown. METHODS: Metaphase comparative genomic hybridization (CGH) was used to identify chromosomal aberrations in 29 tumor specimens. Two tumors were investigated using genome DNA microarrays. RESULTS: Three hundred eleven chromosomal gains and losses were detected by metaphase-CGH. Frequent chromosomal gains were detected at 4q, 6, 12, and X, while frequent chromosomal losses affected regions of chromosomes 8, 16, 17, 22, and Y. Genome DNA microarray analysis in 2 tumors of the series confirmed chromosomal aberrations, detected by metaphase-CGH, and indicated genes such as AURKA (20q13.2) not being recognized by metaphase-CGH. CONCLUSION: Metaphase-CGH results confirmed numerous chromosomal aberrations in juvenile angiofibromas. The most frequent aberrations affected sex chromosomes. Further consensus regions of chromosomal aberrations were detected at 4q, 6, 8, 12, 16, 17, and 22. AURKA and MDM2 were identified as interesting novel amplified genes in juvenile angiofibromas. PMID- 17120310 TI - Retropharyngeal node metastasis from papillary thyroid carcinoma. AB - BACKGROUND: Papillary thyroid carcinomas commonly metastasize to paratracheal and jugular lymph nodes. Metastasis to the retropharyngeal node is rare for this tumor. METHODS: Five patients underwent surgical treatment for metastasis of thyroid papillary carcinoma to the retropharyngeal lymph nodes that presented as a parapharyngeal or retropharyngeal mass. All patients had a history of total or subtotal thyroidectomy as their initial treatment. Among them, 3 patients had undergone ipsilateral modified radical neck dissection at their initial treatment. The other 2 patients had a history of bilateral or ipsilateral modified neck dissection for their subsequent cervical lymph node metastases. RESULTS: Metastatic retropharyngeal nodes were successfully resected via transcervical approach in all patients. Although aspiration and difficulty in swallowing were observed in 2 patients after surgical treatment for metastatic retropharyngeal nodes, these complications spontaneously resolved within a few months. CONCLUSIONS: This study suggests that neck dissection and/or metastatic cervical lymph nodes might alter the direction of lymphatic drainage to the retrograde fashion, resulting in the unusual metastasis to the retropharyngeal lymph nodes. Although the cases described here are rare, metastasis to the retropharyngeal node should be considered at the follow-up for thyroid papillary carcinoma. Because these metastases will be missed by routine ultrasonography of the neck, periodic CT scan or MRI is recommended for follow-up, especially for patients with a history of neck dissection. PMID- 17120311 TI - Screening for local and regional cancer recurrence in patients curatively treated for laryngeal cancer: definition of a high-risk group and estimation of the lead time. AB - BACKGROUND: All patients treated for laryngeal cancer are offered the same follow up schedule to detect asymptomatic locoregional recurrences. In this study, we evaluated the prognostic profile of patients for cancer recurrence and estimated the lead time. METHODS: A cohort study was performed between 1990 and 1995. Cox proportional hazards model was used to analyze the prognostic factors. The effect of altering the follow-up for asymptomatic recurrence detection was determined after estimating the lead time. RESULTS: The variables cT classification, smoking, and histologic grade proved to be prognostic factors. The risk of locoregional failure was 15% in the low-risk group versus 29% in the high-risk group. The estimated lead time was 2 to 4 weeks. CONCLUSION: Risk profiles for locoregional relapse were defined. Intensifying the follow-up schedule is not advisable because the lead time is very short. An excessively high number of routine visits would have to be performed to increase the detection rate for asymptomatic recurrences. PMID- 17120312 TI - Sentinel node in head and neck cancer: use of size criterion to upstage the no neck in head and neck squamous cell carcinoma. AB - BACKGROUND: Anatomical imaging tools demonstrate poor sensitivity in head and neck squamous cell carcinoma (HNSCC) patients with clinically node-negative necks (cN0). This study evaluates nodal size as a staging criterion for detection of cervical metastases, utilizing sentinel node biopsy (SNB) and additional pathology (step-serial sectioning, SSS; and immunohistochemistry, IHC). METHODS: Sixty-five patients with clinically N0 disease underwent SNB, with a mean of 2.4 nodes excised per patient. Nodes were fixed in formalin, bisected, and measured in 3 axes before hematoxylin-eosin staining. Negative nodes were subjected to SSS and IHC. SNB-positive patients underwent modified radical neck dissection. RESULTS: Maximum diameter was larger in levels II and III (13.1 and 13.2 mm) when compared with level I (10.5 mm; p = .004, p = .018), while minimum diameter was constant. Positive nodes were larger than negative nodes (p = .007), but nodes found positive by SSS/IHC were not significantly larger than negative nodes for either measurement (p = .433). Sensitivity and specificity were poor for all measurements. CONCLUSIONS: Nodal size is an inaccurate predictor of nodal metastases and should not be regarded as an accurate means of staging the clinically N0 neck. PMID- 17120313 TI - [Homocysteine--still standing at the end of round ten?]. PMID- 17120315 TI - Consultation-liaison nurses meet. PMID- 17120320 TI - My doctor suggested that I get more exercise. I am considering joining a gym, but how can I cut costs? PMID- 17120321 TI - Institute practices reproductive medicine--and Catholicism. PMID- 17120322 TI - Multiscale dynamics of the cell envelope of Shewanella putrefaciens as a response to pH change. AB - The bacterial surface properties of gram-negative Shewanella putrefaciens were characterized by microbial adhesion to hydrocarbons (MATH), adhesion to polystyrene dishes, and electrophoresis at different values of pH and ionic strength. The bacterial adhesion to these two apolar substrates shows significant variations according to pH and ionic strength. Such behavior could be partly explained by electrostatic repulsions between bacteria and the solid or liquid interface. However, a similar trend was also observed at rather high ionic strength where electrostatic interactions are supposed to be screened. The nanomechanical properties at pH 4 and 10 and at high ionic strength were investigated by using atomic force microscopy (AFM). The indentation curves revealed the presence of a polymeric external layer that swells and softens up with increasing pH. This suggests a concomitant increase of the water permeability and so did of the hydrophilicity of the bacterial surface. Such evolution of the bacterial envelope in response to changes in pH brings new insight to the pH dependence in the bacterial adhesion tests. It especially demonstrates the necessity to consider the hydrophobic/hydrophilic surface properties of bacteria as not univocal for the various experimental conditions investigated. PMID- 17120323 TI - Isolation of microbial DNA by newly designed magnetic particles. AB - Carboxyl group-containing magnetic nonporous poly(2-hydroxyethyl methacrylate-co glycidyl methacrylate) (P(HEMA-co-GMA)) microspheres and cobalt ferrite nanoparticles modified with alginic acid (natural carboxylic polysaccharide) were used for isolation of microbial DNA of lactic acid bacteria (LAB) from dairy products, lyophilised cell cultures, and bacterial colonies grown on hard media, and Trichophyton fungi DNA from lyophilised cells. DNA from the samples with lysed cells was reversibly adsorbed to the particles in the presence of high poly(ethylene glycol) (PEG 6000) and sodium chloride concentrations. The optimal final PEG and NaCl concentrations were 9.1 wt.% and 2.0 M, respectively. The adsorbed DNA was released from the particles in low ionic strength TE buffer. The quality of isolated DNA was checked by PCR amplification. Moreover, PCR amplicons were isolated on cobalt ferrite nanoparticles modified with alginic acid and checked by restriction analysis. PMID- 17120324 TI - Geriatrics and the IOM: did report forget the elderly? PMID- 17120325 TI - The birth of the NETT: NIH-funded network will launch emergency neurological trials. PMID- 17120326 TI - National Highway Traffic Safety Administration (NHTSA) notes. A social norms strategy to reduce impaired driving among 21- to 34-year-olds. PMID- 17120327 TI - Killing controversy. AB - How do neutrophils kill pathogens? A 1967 paper by Seymour Klebanoff provided a neat answer. But far from being resolved, the question still provokes vehement arguments. PMID- 17120328 TI - Bibliography. Current world literature. Transfusion medicine. PMID- 17120329 TI - [Diseases of thyroid - evaluation of the thyroid stimulating hormone (TSH)]. PMID- 17120330 TI - Don't forget the medical directors. PMID- 17120331 TI - Ovary removal linked to increased risk of dementia. PMID- 17120332 TI - Chronic subdural hematoma. Bleeding on the brain. PMID- 17120333 TI - I'm wondering about omega-3 fatty acids. Many times you've stated that fatty, cold-water fish--such as salmon, tuna, mackerel and herring--are good sources of omega-3s. What about freshwater game fish? PMID- 17120334 TI - By the way, doctor. I was vaccinated against Lyme disease in 2000. Do I still need to be careful about tick bites? PMID- 17120335 TI - Authors' reply on Cochrane reviews v industry supported meta-analyses. PMID- 17120336 TI - Hippocrates' tree. PMID- 17120337 TI - Women on methadone. PMID- 17120338 TI - Imaging in general practice. PMID- 17120339 TI - Making CO2 cryotherapy more economical. PMID- 17120340 TI - Apples, oranges and unknown fruit. AB - For many microbiologists their ultimate goal is to understand microbial life at the whole-cell level. Here, Fred Neidhardt gives his view that attention must be paid to the growth conditions for this goal to be realized. PMID- 17120341 TI - Colonic irritation. PMID- 17120342 TI - And the next Director-General of WHO is ... PMID- 17120343 TI - South Africans hope for a new era in HIV/AIDS policies. PMID- 17120344 TI - AAGP issues principles of care for AD-related dementia. PMID- 17120345 TI - Modeling the pregnant woman in driving position. AB - Despite motor vehicle crashes being the leading cause of traumatic fetal morbidity, only a few researches have tried to study the automobile crashes on pregnant women. The possible negative effect of the restraint systems and the injuries mechanisms involved in car crashes with pregnant women are therefore still poorly understood. In this context, the aim of this study is to develop a numerical model of the whole human body with a gravid uterus, in order to investigate car crash scenarios and to evaluate alternative security systems to improve protection of both the woman and the fetus. A 3D reconstruction based on a set of MRI images led us to a good spatial representation of the pregnant woman in driving position. The anatomical precision will make progress possible in the field of traumatology of the pregnant woman. PMID- 17120346 TI - Performance and muscle oxygenation during isometric exercise and recovery in children with congenital heart diseases. AB - This study investigated performance, muscle oxygen saturation (StO2), and blood volume (BV) in patients with congenital heart diseases (CHD) and healthy children during and following sustained exercise. Maximal volunteered contraction (MVC) and endurance at 50 % of MVC (time to exhaustion, Tlim) of the knee extensor were measured in nine patients with CHD and 14 healthy control children. Near infrared spectroscopy was used to evaluated StO2 and BV in vastus lateralis. The drop in muscle oxygen saturation (D(mO2)), half time of recovery (T(SR)), and recovery speed to maximal oxygen saturation (Rs) were analyzed. Patients with CHD showed lower MVC (101.0 +/- 6.2 vs. 125.5 +/- 7.4 N x m, p < 0.01) and Tlim (67.0 +/- 7.5 vs. 127.5 +/- 11.1 s, p < 0.001) than control children. StO2 and BV values in both groups were similar at rest and decreased at the onset of contraction. D(mO2) was larger in patients, which reflected pronounced deoxygenation. During recovery, the patients exhibited a longer TSR (25.2 +/- 2.1 vs. 18.4 +/- 2.0 s, p < 0.05) and R(S) (64.6 +/- 5.5 vs. 42.7 +/- 4.6 s, p < 0.01) than control children. We concluded that reduced strength and endurance in patients with CHD were associated with an impairment of StO2 and BV, and a slower reoxygenation during recovery. PMID- 17120347 TI - Exhaled nitric oxide and airway caliber during exercise-induced bronchoconstriction. AB - Data on the relationship between exercise-induced bronchoconstriction (EIB) and exhaled nitric oxide (NO) in adult patients with asthma are controversial. It is unclear whether endogenous NO may act as either a protective or stimulatory factor in the airway response to exercise or whether changes in exhaled NO simply reflect acute narrowing of the airway. The aim of this study was to assess the changes in the fraction of exhaled nitric oxide (FE(NO)) before and after exercise challenge in patients with asthma and to analyze the relationship between FE(NO) and airway obstruction. Twenty-five non-smoking, steroid-naive, atopic, adult patients with mild persistent asthma and 12 non-smoking, nonatopic, healthy subjects (control group) performed an exercise challenge on a cycloergometer, with monitored ventilation. FEV1 and FE(NO) were measured at baseline and 1, 5, 10, 15 and 20 minutes after the exercise challenge. Eleven of the asthmatic patients had exercise-induced bronchoconstriction (EIB group) and the remaining 14 did not (non-EIB group). Baseline FE(NO) was higher in the EIB and non-EIB asthmatic groups than in the control group. In the EIB group, FE(NO) was significantly lower 5, 10 and 15 minutes after exercise, and the changes in FE(NO) correlated with variation in FEV1 10 and 15 min after exercise. A significant correlation between baseline FE(NO) and maximal post-exercise decrease in FEV1 was found in asthmatic patients (EIB group). In conclusion, exhaled nitric oxide levels transiently decrease during exercise-induced bronchoconstriction in adult patients with asthma. Baseline FE(NO) might predict the airway obstruction resulting after exercise. PMID- 17120348 TI - [The ARCHIVOS archive, 2005: and overview of research published in Archivos De Bronconneumologia]. PMID- 17120349 TI - Seasonal patterns and preterm birth: a systematic review of the literature and an analysis in a London-based cohort. AB - OBJECTIVE: The objectives of this study included a systematic review of the countries in which a seasonal pattern of preterm birth has been reported and an analysis on the seasonal variability of preterm birth in a London-based cohort. DESIGN: Cross-sectional study. SETTING: Eighteen maternity units in a London health region from 1988 to 2000. POPULATION: The study population comprised 482,765, live singleton births born after 24 weeks of gestation and weighing more than 200 g. METHODS: Systematic review and secondary analysis of seasonality over 13 years of births from the St Mary's Maternity Information System (SMMIS). MAIN OUTCOME MEASURE: Annual patterns of preterm birth and a comparison of risk by seasons. RESULTS: Three studies from developing countries and three from developed countries reported a seasonal pattern of preterm birth. One study from the USA reported no seasonal pattern of preterm birth. No British studies were located. Rates of preterm birth in developed countries were highest twice a year (once in winter and again in summer). In London (SMMIS data set), however, preterm births peaked only once a year, in winter. Babies born in winter were 10% more likely to be preterm compared with those born in spring (OR 1.10, 95% CI 1.07-1.14). CONCLUSION: Establishing a seasonal pattern of birth can have important implications for the delivery of healthcare services. Most studies from both developed and developing countries support the existence of preterm birth seasonality. This study has shown that the seasonality of preterm births in this London-based cohort differs from other developed countries that have previously reported a seasonal pattern of preterm birth. PMID- 17120350 TI - Therapy for methicillin-resistant Staphylococcus aureus. PMID- 17120351 TI - Therapy for methicillin-resistant Staphylococcus aureus. PMID- 17120352 TI - Alendronate or alfacalcidol in glucocorticoid-induced osteoporosis. PMID- 17120353 TI - Re: Effect of resin-removal methods on enamel and shear bond strength of rebonded brackets. PMID- 17120354 TI - Re: The effects of first premolar extractions on third molar angulations. PMID- 17120355 TI - Re: Midfacial morphology in adult unoperated complete unilateral cleft lip and palate patients. PMID- 17120356 TI - DeltaFosB: a molecular gate to motivational processes within the nucleus accumbens? PMID- 17120357 TI - Unknown case: part II. Soft tissue herniation through a thoracic laminectomy defect. PMID- 17120358 TI - Long-term fate of neural precursor cells following transplantation into developing and adult CNS. AB - Successful strategies for transplantation of neural precursor cells for replacement of lost or dysfunctional CNS cells require long-term survival of grafted cells and integration with the host system, potentially for the life of the recipient. It is also important to demonstrate that transplants do not result in adverse outcomes. Few studies have examined the long-term properties of transplanted neural precursor cells in the CNS, particularly in non-neurogenic regions of the adult. The aim of the present study was to extensively characterize the fate of defined populations of neural precursor cells following transplantation into the developing and adult CNS (brain and spinal cord) for up to 15 months, including integration of graft-derived neurons with the host. Specifically, we employed neuronal-restricted precursors and glial-restricted precursors, which represent neural precursor cells with lineage restrictions for neuronal and glial fate, respectively. Transplanted cells were prepared from embryonic day-13.5 fetal spinal cord of transgenic donor rats that express the marker gene human placental alkaline phosphatase to achieve stable and reliable graft tracking. We found that in both developing and adult CNS grafted cells showed long-term survival, morphological maturation, extensive distribution and differentiation into all mature CNS cell types (neurons, astrocytes and oligodendrocytes). Graft-derived neurons also formed synapses, as identified by electron microscopy, suggesting that transplanted neural precursor cells integrated with adult CNS. Furthermore, grafts did not result in any apparent deleterious outcomes. We did not detect tumor formation, cells did not localize to unwanted locations and no pronounced immune response was present at the graft sites. The long-term stability of neuronal-restricted precursors and glial restricted precursors and the lack of adverse effects suggest that transplantation of lineage-restricted neural precursor cells can serve as an effective and safe replacement therapy for CNS injury and degeneration. PMID- 17120359 TI - Informed consent in gerontology. AB - The aim of this study was to assess the use and adequacy of informed consent in research involving the elderly in Brazil. Using a reading index, we observed that in 83% of informed consent forms, the text was considered difficult, and demanded a higher schooling level than that presented by the subjects. Whereas 100% of the investigators considered the text in informed consent forms accessible, 75% of the subjects considered it hard to understand. This difference was statistically significant. 94% percent of the elderly participating in research protocols made the decision to participate in the study before reading the term of consent. More attention should be given both to the writing of informed consent forms and to the entire informed consent process, which in gerontology research, should be reviewed at each encounter with study participants. PMID- 17120360 TI - Ethical challenges of human genome diversity research. PMID- 17120361 TI - A historical overview of the developing medical ethics culture in the new Jewish settlement in Israel during the years 1840-1914. PMID- 17120362 TI - NY: LPN refused a transfer to night shift: unemployment benefits denied for 'good cause'. Skinner V. Nathan Littauer Hospital, No. 500149 (N.Y.App.Div. 09/14/2006) -NY. PMID- 17120363 TI - Low-down on link between bone-builders and jaw damage. PMID- 17120364 TI - What's in a name? Perhaps help for tendon pain. PMID- 17120365 TI - Sorting out advances in breast cancer treatment. PMID- 17120366 TI - Itching to find a cure for...itching. PMID- 17120367 TI - Seeing double? See a doctor. PMID- 17120368 TI - My wife is facing a long recovery confined first to bed, and then to a wheelchair. Is there any way to prevent bedsores? PMID- 17120369 TI - Doctors warn of powerful and resistant TB strain. PMID- 17120370 TI - Winners and losers: routine HIV testing is on the cards. Who will it really help? PMID- 17120371 TI - To know or not to know...would routine HIV testing help stem the epidemic, or spell big trouble for vulnerable individuals? PMID- 17120372 TI - The illness industry. PMID- 17120373 TI - Knowing sin: making sure good science doesn't go bad. PMID- 17120374 TI - [Acral melanoma in situ]. PMID- 17120375 TI - Evolution for John Doe: pictures, the public, and the Scopes trial debate. PMID- 17120376 TI - Allotments from federal employees. Interim rule with request for comments. AB - The Office of Personnel Management (OPM) is issuing interim regulations dealing with the use of OPM's allotment authority to allow for pretax salary reductions as part of OPM's flexible benefits plan. Using an allotment from an employee's pay to the employing agency allows certain payments (e.g., employee health insurance premiums, contributions to a flexible spending arrangement, and contributions to a health savings account) to be paid with pretax dollars, as provided under section 125 of the Internal Revenue Code. In addition, these regulations include certain policy clarifications and changes to make the regulations more readable. PMID- 17120377 TI - Rethinking the failure of Fundamentalist political antievolutionism after 1925. PMID- 17120378 TI - Suspension of enrollment in the Federal Employees Health Benefits (FEHB) Program for Peace Corps volunteers. Final rule. AB - The Office of Personnel Management is issuing a final regulation to allow Peace Corps volunteers who are FEHB Program enrolled annuitants, survivors, and former spouses to suspend their FEHB enrollments and then return to the FEHB Program during the Open Season, or return to FEHB coverage immediately, if they involuntarily lose health benefits coverage under the Peace Corps. The intent of this final rule is to allow these beneficiaries to avoid the expense of continuing to pay FEHB Program premiums while they have other health coverage as Peace Corps volunteers, without endangering their ability to return to the FEHB Program in the future. PMID- 17120379 TI - Exemption of work activity as a basis for a continuing disability review. Final rules. AB - We are publishing these final rules to amend our regulations to carry out section 221(m) of the Social Security Act (the Act). Section 221(m) affects our rules for when we will conduct a continuing disability review if you work and receive benefits under title II of the Act based on disability. (We interpret this section to include you if you receive both title II disability benefits and title XVI (Supplemental Security Income (SSI)) payments based on disability.) It also affects our rules on how we evaluate work activity when we decide if you have engaged in substantial gainful activity for purposes of determining whether your disability has ended. In addition, section 221(m) of the Act affects certain other standards we use when we determine whether your disability continues or ends. We are also amending our regulations concerning how we determine whether your disability continues or ends. These revisions will codify our existing operating instructions for how we consider certain work at the last two steps of our continuing disability review process. We are also revising our disability regulations to incorporate some rules which are contained in another part of our regulations and which apply if you are using a ticket under the Ticket to Work and Self-Sufficiency program (the Ticket to Work program). In addition, we are amending our regulations to eliminate the secondary substantial gainful activity amount that we currently use to evaluate work you did as an employee before January 2001. PMID- 17120380 TI - Rules for the issuance of work report receipts, payment of benefits for trial work period service months after a fraud conviction, changes to the student earned income exclusion, and expansion of the reentitlement period for childhood disability benefits. Final rules. AB - We are revising our rules to reflect and implement sections 202, 208, 420A, and 432 of the Social Security Protection Act of 2004 (the SSPA). Section 202 of the SSPA requires us to issue a receipt each time you or your representative report a change in your work activity or give us documentation of a change in your earnings if you receive benefits based on disability under title II or title XVI of the Social Security Act (the Act). Section 208 changes the way we pay benefits during the trial work period if you are convicted by a Federal court of fraudulently concealing your work activity. Section 420A changed the law to allow you to become reentitled to childhood disability benefits under title II at any time if your previous entitlement to childhood disability benefits was terminated because of the performance of substantial gainful activity. Section 432 changes the way we decide if you are eligible for the student earned income exclusion. We will also apply the student earned income exclusion when determining the countable income of an ineligible spouse or ineligible parent. We are also changing the SSI student policy to include home schooling as a form of regular school attendance. PMID- 17120381 TI - Revised medical criteria for evaluating visual disorders. Final rules. AB - We are revising the criteria in the Listing of Impairments (the listings) that we use to evaluate claims involving visual disorders. We apply these criteria when you claim benefits based on disability under title II and title XVI of the Social Security Act (the Act). The revisions reflect our program experience and advances in medical knowledge, treatment, and methods of evaluating visual disorders. PMID- 17120382 TI - Glycemic control and treatment satisfaction in Saudi diabetic children on insulin pump therapy. PMID- 17120383 TI - Urban stress and health in developing countries: development and validation of a neighborhood stress index for India. AB - Stress caused by chronic difficulties encountered by people residing in poor urban neighborhoods is associated with health problems and disease in developed countries, but the relationship between neighborhood stress and health in developing nations, such as India, has not been assessed. In this study, the authors administered the City Stress Inventory, a self-report measure assessing stress experienced as a function of environmental conditions unique to living in large cities that was validated in the United States, to 163 high school students in New Delhi, India. Components of urban stress in India, with some modifications, appear to be similar to components of urban stress reported by adolescents in the United States. Urban stress was predictive of high blood pressure as reported by the adolescents 'parents. In addition, urban stress also predicted health habits, such as chewing tobacco and alcohol use, and psychosocial characteristics, such as hostility. Adolescents' reports of parental stress concerning money and social pressures were also associated with city stress. The current study indicates that the City Stress Inventory is valid in an Indian sample and is predictive of health problems. PMID- 17120385 TI - Smoking cessation 2: behavioral treatments. AB - The authors' purpose of this portion of a 3-part review of interventions for tobacco dependence is to present evidence regarding the efficacy of behavioral treatments for smoking cessation. The present review includes evidence evaluated in the Treating Tobacco Use and Dependence Clinical Practice Guideline (the Guideline) (MC Fiore, WC Bailey, SJ Cohen, et al, 2000) as well as evidence published subsequent to the Guideline. A wide range of effective behavioral treatments for smoking cessation is available. These interventions come in a variety of formats and intensities, can be used by all types of clinicians in a variety of settings, and are appropriate for different types of smokers. The authors have organized recommendations in 2 broad domains that account for much of the variability across behavioral interventions, treatment content, and structure. Finally, the authors discuss key directions for future research. PMID- 17120384 TI - Perceived risk and worry about prostate cancer: a proposed conceptual model. AB - Prostate cancer is one of the most common forms of cancer among American men, and worry about the disease has psychological, behavioral, and biological consequences. To better understand prostate cancer-specific worry, the authors tested a model of the interrelationships among family history of prostate cancer; perceived risk of and worry about prostate cancer; and perceived risk of and worry about other diseases. Men who attended prostate cancer-screening appointments at a general urology practice (n=209) were given a brief anonymous self-report measure. Structural equation modeling (LISREL) results indicated: (1) perceived risk of prostate cancer mediated the relationship between family history of prostate cancer and prostate cancer worry; (2) perceived risk of other diseases increased perceived risk of prostate cancer; and (3) prostate cancer worry and increased other disease worry. PMID- 17120386 TI - How to improve practice communication. PMID- 17120387 TI - A technique for surgical mandibular exostosis removal. AB - Exostosis, a slow-growing, benign bony outgrowth, is a common clinical finding and not usually a concern for patients. However, when removable prosthetics must sit either adjacent to or over these areas, pressure, food abrasion, ulceration, or limited tongue space can occur. This article describes a surgical technique for excision of exostosis through a case presentation. An 86-year-old woman had soft-tissue irritation caused by abrasion from food in the buccal posterior right quadrant. The removal of the exostosis is illustrated through the use of a device that serves as an alternative to a scalpel, offering a safe, predictable outcome. PMID- 17120388 TI - Resin-based composite coronal augmentation of diminutive lateral incisors: an update. AB - The 2 cases presented here document methods of enlarging diminutive lateral incisors using bonded resin-based composite materials. Former methods are reviewed, and descriptions of a matrix strip stabilization technique and the use of a self-etching adhesive agent are included. PMID- 17120389 TI - The effect of temperature on self-etching adhesive penetration. AB - The purpose of this study was to investigate the penetration of an aggressive self-etching adhesive system at refrigerated and room temperatures into ground and unground enamel surfaces. Thirty extracted human teeth were used to measure adhesive penetration into enamel by light microscopy analysis (x400). The unground enamel surfaces were cleaned with pumice and water using a rotary dental brush. For each specimen, part of the unground enamel was manually ground and part was kept intact. A self-etch adhesive was evaluated for its ability to penetrate ground and unground enamel surfaces at room temperature (25 degrees C), at 30 minutes after removal from the refrigerator, and immediately after removal from the refrigerator (6 degrees C). Data were analyzed using variance and the Tukey test, which revealed significant differences in length of penetration of this material when applied on ground and unground enamel surfaces and between the different temperatures used (P > .05). The self-etching system used in this study had significantly lower penetration into unground enamel and at 6 degrees C (P < .05). No statistical difference was found between the interactions of these factors. It was concluded that the self-etching system produced the best penetration into ground enamel surface at room temperature (25 degrees C) and at 30 minutes after removing the specimens from the refrigerator. PMID- 17120390 TI - The effect of fibronectin and a bone xenograft on regenerative treatment: a feasibility study. AB - The purpose of this study was to evaluate the ability of fibronectin to augment the regenerative effects of a bovine-derived xenograft in human periodontal defects. Using a parallel arm, randomized double-blind design, 24 patients with an intrabony defect or a Class II furcation defect were randomly assigned to either the experimental group (xenograft plus fibronectin) or the control group (xenograft without fibronectin). Probing attachment level, pocket depth, and gingival recession were measured at baseline and at 12 months after surgery. Both treatment modalities resulted in attachment gain and pocket depth reduction compared with baseline values. Changes in clinical attachment were not significantly different between the groups (gain of 1.5 mm +/- 1.1 mm in the experimental group and 1.3 mm +/- 1.4 mm in the control). Pocket depth reduction was greater in the control (2.3 mm +/- 1.2 mm) than in the experimental group (2.1 mm +/- 1.9 mm). Gingival recession also was greater in the control (0.9 mm +/- 0.6 mm) than in the experimental group (0.6 +/- 1.5 mm). Subtraction radiography revealed no significant differences between the groups when measuring changes in the distance between the cementoenamel junction and the crest of the bone or in the estimated gain in mineralized tissue mass. It was concluded that a significant difference between the regenerative treatment modalities could not be demonstrated within the limitations of the study. Fibronectin appears to have a stabilizing or proliferative effect on the gingival soft tissue by promoting less postoperative gingival recession. PMID- 17120391 TI - Direct composite resin restorations in posterior teeth. AB - Direct adhesive composite resin restorations are being used extensively. A proper understanding of the technique and its indications has contributed to the widespread use of this restorative approach. Recent industrial developments brought novel technologies to dentistry including the introduction of nanofillers to the restorative procedures. This article reviews some features of this new type of composite resin, discusses its advantages over other materials, and presents an application technique for restoration of posterior teeth through a clinical case report. PMID- 17120392 TI - Applicable research in practice: understanding the hydrophilic and flow property measurements of impression materials. AB - The flow properties and hydrophilicity of an impression material are key factors that affect its performance. This article details in vitro studies comparing these properties in 1 polyether and several vinyl polysiloxane light-body impression materials. The first series of studies examined the materials' flow properties used in a "shark fin" measurement procedure to determine which exhibited superior flow characteristics. The second series of studies reviewed the hydrophilic properties of the materials. Video analysis was used to record contact angle measurements at the early- and late-stage working times. Results showed 1 polyether material to be more hydrophilic. Applying this knowledge to practice, the authors present a clinical case in which a polyether's superior flow and quality of detail were used to make impressions for a patient receiving 8 single-unit zirconia crowns. PMID- 17120393 TI - Large abdominal mass due to a giant uterine leiomyoma. PMID- 17120394 TI - Early referral for chronic kidney disease: good for those who need it, but who are they? PMID- 17120395 TI - Education scholarship. PMID- 17120396 TI - Gulf War Syndrome: the final chapter? PMID- 17120397 TI - Limitations of estimating glomerular filtration rate from serum creatinine in the general population. AB - OBJECTIVE: To compare estimated glomerular filtration rate (GFR) in the general population on the basis of equations derived from different subsets of the general population. PARTICIPANTS AND METHODS: Adults (ages _45 years) were randomly selected from 1997 to 2000 from the Olmsted County, Minnesota, population and had their serum creatinine levels measured. The GFR was estimated using previously reported equations derived from a sample of patients with chronic kidney disease (CKD), a sample of healthy persons, and the combined samples. Serum creatinine was measured with the same assay used to derive these equations. RESULTS: Of 4203 subjects, 2042 (47% participation rate) were enrolled and studied. Serum samples from 1982 subjects were used to measure creatinine levels. The prevalence of a reduced estimated GFR (<60 mL/min per 1.73 m2) was 12% (95% confidence interval [CI], 10%-13%) based on an equation derived with all CKD patients, and this finding was similar to prior reports. However, the prevalence of a reduced estimated GFR was 5.7% (95% CI, 4.8%-6.8%) based on an equation derived with both CKD patients and healthy persons and 0.2% (95% CI, 0.1%-0.5%) based on an equation derived with all healthy persons. Women had a higher risk of reduced estimated GFR according to an equation derived with all CKD patients, but men had a higher risk with an equation derived with both CKD patients and healthy persons. PMID- 17120398 TI - A multicenter study of burnout, depression, and quality of life in minority and nonminority US medical students. AB - OBJECTIVE: To determine the well-being of minority medical students in a multicenter sample of US medical students. PARTICIPANTS AND METHODS: All 1098 medical students at 3 medical schools in Minnesota were surveyed in April 2004. Validated instruments were used to assess burnout, depression, and quality of life (QOL). Students were also asked about the prevalence of significant personal life events in the previous 12 months and strategies used to cope with stress. RESULTS: Although symptoms of depression and overall burnout were similar among minority and nonminority students, minority students were more likely to have a low sense of personal accomplishment (P=.02) and lower QOL In a number of domains (all P< or =.05). These differences persisted on multivariate analysis that controlled for demographic characteristics and recent life events. Minority students were also more likely to have a child (P=.01), originate from outside Minnesota (P<.001), and experience a major personal Illness in the last 12 months (P=.03). CONCLUSION: As a group, the minority medical students in this survey had a lower sense of personal accomplishment and QOL than nonminority students. Additional studies are needed to provide insight regarding the causes of these inequities and the unique challenges faced by minority medical students. Efforts to improve minority students' well-being, QOL, and learning experience may help prevent attrition among minority medical students and promote diversification in the physician workforce. PMID- 17120399 TI - The benefits of pass-fail grading on stress, mood, and group cohesion in medical students. AB - OBJECTIVE: To objectively measure the effect of a pass-fail grading system on stress, mood, group cohesion, and test anxiety in medical students. PARTICIPANTS AND METHODS: Beginning with the class of 2006, the Mayo Medical School, Rochester, Minn, changed the grading system for first-year courses from a 5 interval grading system to a pass-fail grading system. Students in the previous class of 2005, who were graded using a 5-interval system during their first year of medical school, were compared with students in the class of 2006. Using a prospective study design, the 2 groups were compared at the end of both the first year and the second year of medical school on the Perceived Stress Scale, Profile of Mood States, Perceived Cohesion Scale, Test Anxiety Inventory, and (after year 2) the United States Medical Licensing Examination Step 1. Data collection occurred in 2002 and 2003 with the class of 2005 and in 2003 and 2004 with the class of 2006. RESULTS: Students graded with the pass-fail system had less perceived stress (median, 15.0 vs 21.0; P-.01) and greater group cohesion (median, 34.5 vs 30.0; P=.02) at the end of their second year of coursework than their 5-interval graded peers. The pass-fail group had better mood (median, 46.5) than the graded group (median, 64.0), but this difference was not statistically significant (P=.07). No significant differences were found between the 2 groups in test-taking anxiety or in United States Medical Licensing Examination Step 1 board scores. CONCLUSION: Pass-fail grading may reduce stress and increase group cohesion in medical students compared with traditional 5-interval grading. PMID- 17120400 TI - The impact of parental leave on extending training and entering the board certification examination process: a specialty-based comparison. AB - OBJECTIVE: To study the impact of parental leave on extending residency training and the timing of entrance into the specialty board certification examination system. METHODS: From June 3, 2005, through December 28, 2005, primary specialty board policies regarding parental leave, absence from training, and the timing of entrance into the board certification examination process were reviewed for all American Board of Medical Specialties programs with a national enrollment of more than 100 residents. The impact of a 6-week parental leave on extending training and qualifying to enter the board certification examination system was compared among these specialties. RESULTS: All specialty boards studied, except for the American boards of neurosurgery, ophthalmology, psychiatry, neurology, and thoracic surgery, have defined limits on absences from training. The limits on absence from training among the 21 other residency programs studied are generally similar, but important differences exist. These differences include the maximum length of time away from training per year, whether absence from training can accumulate year to year, and whether the length of time away from training is consistent for each postgraduate year of the training program. The impact of a 6 week parental leave on qualifying for the board certification examination system on schedule varies from no impact to delaying entrance for 1 year. CONCLUSION: Specialty board policies regarding absence from training and entering the board certification examination process vary and could influence decisions about family planning, the length of time taken for parental leave, the use of vacation time for parental leave, and resident well-being. PMID- 17120401 TI - A comparison of intensive care unit physician staffing costs at the 3 Mayo Clinic sites. AB - OBJECTIVES: To determine the provider cost of administering intensive care unit (ICU) services, comparing 3 different staffing models for ICU coverage, and to compare the costs of using house staff vs nonphysician providers (NPPs). METHODS: Data were collected on total staff composition and number of beds In ICUs from January 1, 2004, through December 31, 2004, at the 3 Mayo Clinic sites: Rochester, Minn; Jacksonville, Fla; and Scottsdale, Ariz. Institutional or national average staff salaries were used to determine total staffing costs per ICU bed per year at each site. Medicare medical education reimbursements were also taken into account. RESULTS: Costs per ICU bed for physician staffing were $18,630 in Rochester, $37,515 in Jacksonville, and $38,010 in Scottsdale. When NPPs were substituted for house staff, the costs per bed were $72,466 in Rochester, $61,291 in Jacksonville, and $49,909 in Scottsdale. Incremental costs per ICU bed using NPPs were $53,836 in Rochester, $23,776 in Jacksonville, and $11,899 in Scottsdale. CONCLUSION: Use of residents and fellows in ICU staffing at a major tertiary health center is more cost-efficient than use of NPPs. This finding could have Implications for the cost of physician services in nonteaching community hospitals and the methods by which care is provided. PMID- 17120402 TI - Smoking and inflammatory bowel disease: a meta-analysis. AB - OBJECTIVE: To assess whether there is a true effect of smoking on the 2 most prevalent forms of inflammatory bowel disease (IBD): Crohn disease (CD) and ulcerative colitis (UC). METHODS: For this meta-analysis, we searched multiple health care databases, including MEDLINE and EMBASE (January 1980 to January 2006), to examine the relationship between smoking and IBD. Keywords searched included smoking, Inflammatory bowel disease, Crohn's disease, and ulcerative colitis. Data were abstracted using predefined inclusion and exclusion criteria. An odds ratio (OR) was recalculated for each study using the random-effects model, and a combined OR was calculated. RESULTS: A total of 245 articles were obtained through an electronic search of health care databases. Thirteen studies examined the relationship between UC and smoking, whereas 9 examined the relationship between CD and smoking. We found evidence of an association between current smoking and CD (OR, 1.76; 95% confidence interval [CI], 1.40-2.22) and former smoking and UC (OR, 1.79; 95% CI, 1.37-2.34). Current smoking had a protective effect on the development of UC when compared with controls (OR, 0.58; 95% CI, 0.45-0.75). CONCLUSION: This is the first meta-analysis, to our knowledge, to evaluate the relationship between smoking and IBD using accepted quality standards for meta-analysis reporting. Our meta-analyses confirm that smoking is an important environmental factor in IBD with differing effects in UC and CD. By using predefined inclusion criteria and testing for homogeneity, the current analysis provides an estimate of the effect of smoking on both these forms of IBD. PMID- 17120403 TI - eQuality: electronic quality assessment from narrative clinical reports. AB - OBJECTIVE: To evaluate an electronic quality (eQuality) assessment tool for dictated disability examination records. METHODS: We applied automated concept based indexing techniques to automated quality screening of Department of Veterans Affairs spine disability examinations that had previously undergone gold standard quality review by human experts using established quality indicators. We developed automated quality screening rules and refined them iteratively on a training set of disability examination reports. We applied the resulting rules to a novel test set of spine disability examination reports. The initial data set was composed of all electronically available examination reports (N=125,576) finalized by the Veterans Health Administration between July and September 2001. RESULTS: Sensitivity was 91% for the training set and 87% for the test set (P .02). Specificity was 74% for the training set and 71% for the test set (P=.44). Human performance ranged from 4% to 6% higher (P<.001) than the eQuality tool in sensitivity and 13% to 16% higher in specificity (P<.001). In addition, the eQuality tool was equivalent or higher in sensitivity for 5 of 9 individual quality indicators. CONCLUSION: The results demonstrate that a properly authored computer-based expert systems approach can perform quality measurement as well as human reviewers for many quality indicators. Although automation will likely always rely on expert guidance to be accurate and meaningful, eQuality is an important new method to assist clinicians in their efforts to practice safe and effective medicine. PMID- 17120404 TI - Diagnosis and management of pathological laughter and crying. AB - Patients with various neurologic disorders exhibit exaggerated or inappropriate episodes of laughter, crying, or both without an apparent motivating stimulus or in response to stimuli that would not have elicited such an emotional response before the onset of the underlying disease. During these episodes, patients have difficulty controlling their emotional expression according to the contextual information. In contrast, patients with mood disorders have a pervasive and sustained change in their emotional experience and thus exhibit spells of laughter or crying because of an underlying mania or depression. This article focuses on the clinical presentation, diagnosis, prevalence, and proposed pathophysiological mechanisms of and available treatment options for this clinical phenomenon. PMID- 17120405 TI - Late referral of patients with chronic kidney disease: no time to waste. AB - The prevalence of patients with chronic kidney disease (CKD) in the US population is approximately 11%, and because of the increase in life expectancy and in diabetic nephropathy incidence, an exponential increase is predicted for the next decades. During the past decade, evidence that the progression of CKD can be attenuated by a multifactorial therapeutic approach has been increasing. However, a substantial percentage of patients with CKD will have progression to CKD stage V (ie, need for renal replacement therapy). Late referral of these patients (ie, <1 to 6 months before the start of renal replacement therapy) has been shown to be associated with higher mortality, morbidity, and costs. However, up to 64% of patients with CKD are still referred late. This review presents the available data on the epidemiology, causes, and consequences of late patient referral. Furthermore, it offers information to prevent late referral, improve CKD patient care, and change clinical practice. PMID- 17120406 TI - 47-Year-old woman with spells of slurred speech, blurred vision, and loss of consciousness. PMID- 17120407 TI - Transient stress-induced cardiomyopathy with an "inverted takotsubo" contractile pattern. AB - We describe a patient who had transient cardiomyopathy with akinesia of the basal portions of the left ventricle and hyperkinesia of the apex triggered by alcoholic pancreatitis. This case seems to confirm recent publications suggesting a new or variant clinical entity with a clinical presentation similar to that of Takotsubo cardiomyopathy but with an Inverse left ventricular contractile pattern ("Inverted Takotsubo"). This entity could provide clues to the underlying pathophysiology of these syndromes of acute heart injury. PMID- 17120408 TI - Lactic acidosis in patients with neoplasms: an oncologic emergency. PMID- 17120409 TI - "Futile" care in the intensive care unit. PMID- 17120410 TI - President Taft's blood pressure. PMID- 17120411 TI - Frequency of hypertension in patients with gout. PMID- 17120412 TI - [Andrei L'vovich Polenov (1871-1947)]. PMID- 17120413 TI - [Effectiveness of using autologous mono-nuclears of the bone marrow in treatment of patients with ischemic heart disease]. AB - The authors have analyzed their experiences with treatment of 50 patients with ischemic heart disease using transplantation of autologous mono-nuclears of the bone marrow. It was shown that this operation resulted in an improvement of indices of the heart functions and myocardium metabolism. Transplantation of stem cells as mononuclear fraction of the bone marrow is indicated in treatment of different groups of patients: in recurrent diseases after previous operations on the coronary arteries; in patients with distal lesions of the coronary bed; transplantation of autologous stem cells of the bone marrow is expedient simultaneously with coronary artery bypass grafting or coronary angioplasty (stenting). PMID- 17120414 TI - [Prosthesis of mitral valve in mesenchimal dysplasia: anatomo-morphological aspects and technical specificities]. AB - Pathomorphological aspects of acquired mitral defects of the heart caused by manifestations of mesenchimal dysplasia were studied in 100 patients. Special attention is given to important features of surgical strategy when performing mitral valve prosthesis in such patients unlike similar procedures for mitral defects of another etiology. PMID- 17120415 TI - [Modern methods of treatment of inguinal hernias]. AB - Surgical methods of treatment have been analyzed with special reference to wide introduction of new technologies. Altogether there were 4424 operations for inguinal hernias. Traditional plasty of the posterior wall of the inguinal canal with autotissues was used in 2136 patients. Lichtenstein operations were fulfilled on 521 patients, laparoscopic transabdominal preperitoneal hernioplasty -on 1614, complete preperitoneal hernioplasty--on 153 patients. The analysis made has shown that an introduction of modern surgical technologies into clinical practice allowed reduction of the percentage of complications and recurrences. The share of operations by the "tension-free," method was as high as 52%. PMID- 17120416 TI - [Epidemiology of abdominal sepsis]. AB - An analysis of 2376 case histories of patients with surgical infections has shown that in 827 (34.8%) patients the course of main disease was complicated by sepsis. The most frequent causes of the development of sepsis were surgical infections with localization in the abdominal cavity, lungs and mediastinum. Abdominal sepsis was diagnosed in 398 (41.7%) of patients with peritonitis. The annual growth of this category of patients was 8.3%. The outcome of abdominal sepsis was shown to depend on the score number according to scales APACHE II and SOFA, and on the timely operative treatment and beginning of antibacterial therapy. PMID- 17120417 TI - [Assessment of effectiveness of gastroduodenal anastomosis in the surgery of gastric cancer]. AB - An analysis of results of surgical treatment was made on the basis of experiences of performing 260 operations with gastroduodenal anastomosis for distal gastric cancer under conditions of the same hospital for 20 years. Postoperative lethality was 2.69%. Different complications after surgery were noted in 34.2% of the patients. Dumping syndrome after Billroth-1 operations developed in 12.3%. The 5 and 10 year survival after operations among 215 followed-up patients was 65.1% and 24.7% respectively. Original methods of operations were developed and introduced into practice for the improvement of immediate and long-term functional results of the surgery on the stomach. PMID- 17120418 TI - [Principles of treatment of nonparasitic cysts of the liver]. AB - A comparative investigation of different methods of treatment of nonparasitic cysts of the liver was carried out in 104 patients aged from 21 through 74 years with sizes of the cysts from 6 to 21 cm in diameter, divided into three groups. The first group included 50 patients who were subjected to open operations (segmentectomy, bisegmentectomy, fenestration of the cysts with treatment of the residual inner lining with ethanol, diathermocoagulation or cryodestruction). Laparoscopic fenestration and chemical or thermal treatment of the cysts were used in the second group of 29 patients, in the third group consisting of 25 patients a puncture-aspiration method was used under sonographic control. An analysis of the results showed advantages of laparoscopic and puncture methods that gave good immediate and long-term results, less number of postoperative complications and shorter periods of rehabilitation of the patients. PMID- 17120419 TI - [Intravascular laser irradiation of blood in complex treatment of obliterating atherosclerosis of the lower extremity vessels in elderly and senile patients]. AB - The authors describe comparative results of treatment of 60 elderly and senile patients with obliterating atherosclerosis of the lower extremity vessels. In 50 patients treatment with traditional medicines was combined with intravascular laser irradiation of blood, 10 patients were treated with traditional medicines only. The data obtained by questionnaires concerning the patients' state, expert judgment of doctors in charge of the profile department, indices of instrumental examinations of regional hemodynamics (rheovasography of the lower extremity vessels), data of laboratory investigations of morphofunctional state of erythrocytes and hemorheology showed that laser irradiation of blood gave better results of treatment. Its therapeutic effect persisted during 3 months in most patients. PMID- 17120420 TI - [Hemodynamic characteristics and priority of blood refluxes in genesis of trophic ulcers in patients with varicose disease of lower extremities]. AB - An analysis of results of complex examination of 75 patients with varicose disease and trophic ulcers has shown that in 81.4% of the patients the main hemodynamic factor resulting in the development of ulcers was highly intensive total-subtotal reflux of blood along the superficial veins, and in 9.3% it was the highly intensive profound reflux along the posterior tibial veins. In 9.3% of the patients the priority of blood reflux was not established. Trophic ulcers were developing under the influence of low intensity refluxes of blood in the superficial, profound veins and perforators in elderly patients against the background of heart failure. PMID- 17120421 TI - [Comparative clinical assessment of the factors determining the volume of operations for sarcomas of the extremity soft tissues]. AB - In an analysis of data of 526 patients with carcomas of the extremity soft tissues the factors influencing the decision on the volume of surgery were determine. The 5 year survival of patients subjected to amputations was considerably lower than after sparing operations. Amputations were mostly made for local recurrences of the disease, neoplasm larger than 15 cm in diameter, low differentiated sarcomas, with infiltration of the tumor into the bone and/or periosteum and localization of the tumor in distal parts of extremities. The rates of 5 year survival among the patients with malignant tumors of the extremity soft tissues subjected to sparing operations were found to be higher after combined program of treatment (operation + radiation therapy). PMID- 17120422 TI - [Decision on the strategy of treatment of patients with medullary cancer of the thyroid]. AB - The work is based on an analysis of immediate and long-term results of treatment of 138 patients. It was shown that organ-sparing operations in combination with central lymphadenectomy were good for patients with intrathyroid carcinomas. Regional metastases observed by the moment of surgical treatment were a bad prognostic sign. The survival of patients without metastases was similar to that of patients with papillary and follicular cancer of the thyroid gland. PMID- 17120423 TI - [Model of prognosis of outcome of burn trauma on the basis of probit-analysis]. AB - On the basis of probit-analysis of results of treatment of 10,670 burned patients a prognostic model of the trauma outcome was created as a coordinate network. The model is very accurate, sensitive, specific and simple in use that allows it to be applied for prognosis of burn trauma outcomes in the early period after trauma, especially in organization of medical-evacuation work in places of appearance of mass burns. The model is a standard for the assessment of new protocols of treatment and can serve a criterion of the efficiency of work of intensive care units in burn centers. PMID- 17120424 TI - [Anesthesiologist-resuscitator: problems of training and maintenance of professional work]. AB - The problems of psychological disturbances and insufficient profeccional skills amongst anaesthesiologists and intensive care specialists are discussed. The authors emphasized the need for special psychological education during refreshing courses and training period in residency. The official education in clinical residency should be updated in accordance with current view of speciality. It is necessary to use special simulators for better training of manual skill's and evaluation of knowledge more widely. The proper organization of the workplace is very important for successful performance and psychological condition of the personnel. The lack of profeccional Russian-language journals makes necessity for creation of a new textbook and periodical issue in paper and electronic format. PMID- 17120425 TI - [Pharmacoeconomical grounds for decision on a basic hypnotic for general anesthesia in surgical interventions]. AB - A comparative pharmacoeconomical analysis of basic hypnotics has been made in 127 patients based on an assessment of quality of anesthesia using standard scales. The data obtained show the use of propofol and a combination propofol-midazolam to be expedient. These medicines give reliably better results than using propofol H20, sodium thiopental, and a combination of thiopental with droperidol. PMID- 17120426 TI - [Transfacial accesses to tumors of the clivus]. AB - Treatment of 3 patients with malignant tumors of the skull base using transfacial accesses as half maxilotomy by the Kosson access and access with osteotomy of a nasoorbital complex was analyzed that provided a possibility to ablate large and gigantic tumors of the localization in question using the operative microscope and a traditional instruments' kit. PMID- 17120427 TI - [Choice and results of laparoscopic suture of perforated gastroduodenal ulcers]. AB - Results of treatment of 655 patients aged from 17 through 89 years for perforative ulcers of the stomach and duodenum were analyzed retrospectively. Simple suturing of ulcer was performed in 526 (80.3%) patients, laparoscopic methods included. Comparison of general results of treatment of 655 patients with the data of the control group consisting of 390 patients treated at the same hospital before introduction of laparoscopic operations failed to reveal statistically significant differences in frequency of postoperative complications and lethality. The authors defined general somatic, technical and tactical contraindications for laparoscopic operations. PMID- 17120428 TI - [Long-term results of conservative treatment of patients with obliterating atherosclerosis of the lower extremity arteries]. AB - An analysis of long-term results of treatment of 56 patients has shown that a complex program including the intravascular photohemotherapy and a course of a modified method of plasmapheresis is the most effective method in such cases. PMID- 17120429 TI - [Primary-multiple carcinoma of the uterine cervix and stomach in an elderly woman]. PMID- 17120430 TI - [Autotransplantation of the heart as a method of treatment of valvular disease, atriomegalia with pronounced disturbances of hemodynamics]. PMID- 17120431 TI - [Vagotomy in surgical gastroenterology: legends and reality]. AB - The work has shown high effectiveness of vagotomy in a number of diseases. In 2565 patients with perforated duodenal ulcer operation of suturing finished with lethal outcome in 9.2% of cases and recurrent disease in 54% of patients. After vagotomy the results were 1.5% and 14% respectively. In 277 patients with massive bleeding due to erosive gastritis lethality after different operations was 39 46%, after vagotomy--10.0%. After 403 vagotomies for decompensated stenosis of the duodenum atony of the stomach developed in 1.2% of patients, recurrent ulcer appeared in 6%, lethality was 1%. Vagotomy proved to be effective for peptic ulcers of the jejunum (106 patients) in 86% of cases, for chronic indurative pancreatitis (86 patients) in 97% of cases, lethality was 1.2%. PMID- 17120432 TI - [Professor Anatolii Grigor'evich Zemlianoi (1925-1998)]. PMID- 17120433 TI - [Traumatic arterial aneurysms]. PMID- 17120434 TI - [Use of purines in anesthesiology and intensive therapy]. PMID- 17120435 TI - Checking the pulse of water-quality credit trading. PMID- 17120436 TI - Stormwater quality benefits of a porous friction course and its effect on pollutant removal by roadside shoulders. AB - This project has two main objectives; the first is to documents the effect of a porous friction course overlay on the quality of highway stormwater runoff, and the second is to assess pollutant removal of a vegetated area along the side of a road from both conventional and porous pavements. The quantity and quality of stormwater runoff from a four-lane divided highway in the Austin, Texas, area was monitored before and after the installation of a porous friction course (PFC). Observed concentrations of suspended solids and pollutants associated with particulate material were much lower in the runoff from the PFC compared with runoff from the conventional asphalt surface. The vegetated highway shoulder reduced the concentrations of many stormwater constituents from the conventional asphalt surface; however, the concentrations of these constituents in runoff produced by the PFC were so low that the vegetated area produced no further reduction in concentration. PMID- 17120437 TI - Effect of humification processes on polyaromatic hydrocarbons concentration during wastewater treatment. AB - The aim of the present work was to compare the concentration changes of polyaromatic hydrocarbons (PAH) and the course of humification processes during wastewater treatment. Studies of samples from a biological-mechanical wastewater treatment plant in Sosnowiec-Zagorze (Poland) were carried out. Determination of PAH was performed both for wastewater sludge and sludge water. Observations of the course of humification processes for humic acid fractions isolated from sludges were conducted. Analysis of PAH extracted from wastewaters and from sludge was performed by means of high-performance liquid chromatography. Investigations of humification processes were conducted by electron paramagnetic resonance and nuclear magnetic resonance methods. The elementary composition changes in the structure of the extracted humic acids were determined. It was found that polyaromatic hydrocarbons appear during the processes of humification. Their content in water decreased only after the process of sludge aeration; however, sludge water leaving the settlers was PAH-enriched. PMID- 17120438 TI - Simultaneous nitrification and denitrification with anoxic phosphorus uptake in a membrane bioreactor system. AB - The performance of an innovative membrane bioreactor (MBR) process using anoxic phosphorus uptake with nitrification and denitrification for the treatment of municipal wastewater with respect to operational performance and effluent quality is addressed in this paper. The system was operated at steady-state conditions with a synthetic acetate-based wastewater at a hydraulic retention time (HRT) of 12 hours and on degritted municipal wastewater at a total system HRT of 6 hours. The MBR system was able to achieve 99% biochemical oxygen demand (BOD), chemical oxygen demand (COD), and ammonia-nitrogen (NH4(+)-N); 98% total Kjeldahl nitrogen (TKN); and 97% phosphorus removal, producing effluent BOD, COD, NH4+-N, TKN, nitrate-nitrogen, nitrite-nitrogen, and phosphate-phosphorus of <3, 14, 0.2, 0.26, 5.8, 0.21, and <0.01 mg/L, respectively, at the 6-hour HRT. The comparison of the synthetic and municipal wastewater run is presented in this paper. Steady state mass balance on municipal wastewater was performed to reveal some key features of the modified MBR system. PMID- 17120439 TI - Gas phase transport in gravity sewers--A methodology for determination of horizontal gas transport and ventilation. AB - A method was developed for determination of horizontal gas transport and ventilation in gravity sewers. This was achieved by changing the composition of the sewer atmosphere by pulse injection of oxygen gas and subsequently measuring the oxygen concentration in a downstream manhole. Conventional tracer techniques may require sampling and may also affect the environment. The method developed is simple, based on direct monitoring and without environmental or toxic effects. The method was developed based on measurements in an intercepting gravity sewer. The horizontal gas transport processes were quantified by measuring the velocity and dispersion of the gas in the sewer atmosphere. Based on 54 measurements, the gas velocity was found to vary between 0.05 and 0.22 m/s. The coefficients of dispersion were calculated to be in the range 0.05 to 1.1 m2/s. Climatic conditions did not significantly influence the gas phase transport. PMID- 17120440 TI - Influence of chitosan characteristics and environmental conditions on flocculation of anaerobic sludge. AB - The effects of chitosan characteristics (i.e., degree of deacetylation [DD] and molecular weight) and environmental conditions (i.e., ionic strength and pH) on the flocculation of anaerobic sludge were investigated. The results showed that chitosan enhanced the flocculation of sludge, and the flocculation efficiency depended on both the degree of deacetylation and molecular weight. Chitosan with 85%DD was more effective than that with 70%DD, as the former required a lower dose to obtain 90% flocculation at all studied pH values. In addition, low molecular weight chitosan enhanced the flocculation better than high molecular weight chitosan. The increase in ionic strength (up to 0.1 M) of the suspension helped reduce restabilization that occurred when chitosan was overdosed. In general, chitosan has potential to be used as an effective cationic bioflocculant, which is able to function either in acidic or neutral conditions, and very small amounts of chitosan (less than 4 mg/g dried sludge) are required. PMID- 17120441 TI - Biosolids-amended soils: Part I. Effect of biosolids application on soil quality and ecotoxicity. AB - Investigations of potential risk from biosolids generally indicate that land application does not threaten human or ecosystem health, but questions continue to arise concerning the environmental effects of this practice. This research project was initiated to evaluate ecotoxicity resulting from the amendment of soils with biosolids from municipal wastewater treatment plants. Toxicity was evaluated using standard tests, including earthworm mortality, growth, and reproduction; seedling germination and root elongation; microbial respiration; and nematode mortality and reproduction. Nineteen municipal wastewater treatment plants were identified to participate in an initial screening of toxicity, and five were chosen for a more detailed evaluation. In addition, two soils with historically high applications of high-metal biosolids were evaluated. Contaminants examined were zinc, copper, nickel, chromium, arsenic, cadmium, lead, and coplanar polychlorinated biphenyls (PCBs). Single applications had no effect on soil metal concentrations. Coplanar PCBs were not detectable in any of the soils or biosolids. All target organisms were sensitive to reference toxicants. Limited toxicity was observed in a small number of the amended soils, but no patterns emerged. Approximately one-half of the negative effects of biosolids on bioindicators could be attributed to routine properties, such as slight depression of pH and/or elevated salinity. None of the accumulated metal concentrations were excessive, and most would not be considered elevated. These observations suggest that current regulations for application of biosolids to soils are providing adequate ecosystem protection. PMID- 17120442 TI - Biosolids-amended soils: Part II. Chemical lability as a measure of contaminant bioaccessability. AB - Biosolids recycling by amending agricultural soils has increased significantly over the last few decades. The presence of contaminants in small, bioavailable quantities has generated concerns about health threats resulting from accumulation of potential toxins in the food chain. In this study, land application of biosolids was evaluated for environmental risk. Chemical lability tests for metals were used for the test soils and included analyses for water soluble, exchangeable, and metals extractable by the physiologically based extraction test. Chemical extractions detected slight increases in labile metal concentrations for many of the treated soils, particularly those receiving long term applications of 5 years or more. Significantly higher metal concentrations were observed in the soils that had been exposed to biosolids before the U.S. Environmental Protection Agency (Washington, D.C.) 503 Rule (U.S. EPA, 2004) was implemented. PMID- 17120443 TI - Full-scale class A biosolids production by two-stage continuous-batch thermophilic anaerobic digestion at the hyperion treatment plant, Los Angeles, California. AB - The City of Los Angeles Hyperion Treatment Plant (HTP) (California) converted its anaerobic digesters to thermophilic operation to produce Class A biosolids. Phase IV tests demonstrated compliance of a two-stage, continuous-batch process with Alternative 1 of U.S. Environmental Protection Agency 40 CFR Part 503 (U.S. EPA, 1993), which defines the time-temperature requirement for batch treatment (T > or = 56.3 degrees C at 16-h holding). Fecal coliforms, Salmonella sp., viable helminth ova, and enteric viruses were not detected in biosolids in the postdigestion train, including the truck-loading facility and the farm for land application as the last points of plant control where compliance is to be demonstrated. The same results were achieved during Phase V tests, after lowering the second-stage holding temperature to 52.6 degrees C to reduce the elevated methyl mercaptan production that was observed during Phase IV. Hence, the Phase V process complied with Alternative 3 of 40 CFR Part 503. Currently, HTP operates its digesters under the same conditions as tested in Phase V. In 2003, monthly monitoring of the biosolids at the truck-loading facility and the farm for land application demonstrated consistent compliance with Alternative 3. PMID- 17120444 TI - Transport of coliphage PRD1 in a surface flow constructed wetland. AB - A tracer study was conducted in a 3-ha surface flow constructed wetland to analyze transport performance of PRD1, an enteric virus model. The convection dispersion equation (CDE), including a first-order reaction model, adequately simulated transport performance of PRD1 in the wetland under an average hydraulic loading rate of 82 mm/d. Convective velocity (v) and longitudinal dispersion coefficient (D) were estimated by modeling a conservative tracer (bromide) pulse through the wetland. Both PRD1 and bromide were simultaneously added to the entering secondary treated wastewater effluent. The mass of bromide and PRD1 recovered was 76 and 16%, respectively. The PRD1 decay rate was calculated to be 0.3/day. The findings of this study suggest that the CDE model and analytical moment equations represent a suitable option to characterize virus transport performance in surface flow constructed wetlands. PMID- 17120445 TI - A study for evaluation of contaminant transport characteristics through fine grained soil. AB - Transport of soluble toxic substances through porous media lead to some significant geoenvironmental problems, for example, leachate migration from municipal and industrial solid waste resulting from unregulated disposal. Advection, dispersion, diffusion, and decay are reported to be the principal mechanisms in such phenomena. Geotechnical properties of the soil also play a significant role in this deterioration. In the present study, laboratory tests were conducted to formulate an appropriate method for assessment of migration of metal ions, such as nickel, through the soil. Relevant kinetic and process parameters, such as aquifer data, surface area, dielectric constant, pH of zero point charge (pHzpc), and permeability were also studied. One-dimensional mathematical modeling was used to describe the dynamics of the process. The present investigation was carried out at an ash pond site of a thermal power plant situated in West Bengal, India. PMID- 17120446 TI - Removal of aqueous nickel (II) using laterite as a low-cost adsorbent. AB - The present paper describes the laboratory study of laterite as a low-cost adsorbent for removal of aqueous nickel (II). At pH 7 and a temperature of 30 degrees C, a sorbent dose of 15 mg/L resulted in approximately 90% removal of nickel (II) from its initial concentration of 10 mg/L. A maximum removal of 98% of the adsorbate was observed with an adsorbent particle size of 210 micro with the above conditions. Batch kinetics results were described by fitting in a Langmuir isotherm. Helffrich's half-time equation (Helffrich, 1962) has been applied to evaluate the adsorption process. It appears that film diffusion would be the rate-limiting step. The effect of pH on the sorption process was carried out to a value of 8.0. The removal rate of nickel was found to be the function of pH of the reaction mixture. The rate of nickel uptake by laterite with the decrease in pH value has been explained on the basis of aqueous-complex formation and the subsequent acid-base dissociation at the solid-solution interface. PMID- 17120447 TI - Evaluation of the persistence of micropollutants through pure-oxygen activated sludge nitrification and denitrification. AB - The persistence of pharmaceuticals, hormones, and household and industrial chemicals through a pure-oxygen activated sludge, nitrification, denitrification wastewater treatment facility was evaluated. Of the 125 micropollutants that were tested in this study, 55 compounds were detected in the untreated wastewater, and 27 compounds were detected in the disinfected effluent. The persistent compounds included surfactants, fire-retardant chemicals, pesticides, fragrance compounds, hormones, and one pharmaceutical. Physical-chemical properties of micropollutants that affected partitioning onto wastewater solids included vapor pressure and octanol-water partition coefficients. PMID- 17120448 TI - Removal of pyrene and benzo(a)pyrene from contaminated water by sequential and simultaneous ozonation and biotreatment. AB - The removal of pyrene and benzo(a)pyrene from contaminated water by sequential and simultaneous ozonation-bioremediation techniques was investigated. During the sequential treatment, ozonation using 0.5 or 2.5 mg/L ozone was used as a pretreatment process, whereas, during the simultaneous treatment process, ozonation of hydrocarbon-contaminated water at a predetermined duration using 0.5 mg/L ozone was made in the presence of microbial biomass. Ozonation was not beneficial for the removal of pyrene. However, despite a decreased specific biodegradation rate, ozonation improved the overall elimination of benzo(a)pyrene during both treatment processes. The overall removal of benzo(a)pyrene increased from 23 to 91% after exposure of the water to 0.5 mg/L ozone for 30 minutes during the simultaneous treatment process and further to 100% following exposure to 2.5 mg/L ozone for 60 minutes during the sequential treatment mode, demonstrating the benefits of combined ozonation-biological treatment for the removal of polycyclic aromatic hydrocarbons. PMID- 17120449 TI - The increasing need for science to inform policy--more than 30 years passage Clean Water Act. PMID- 17120450 TI - Decomposition of 2-nitrophenol in aqueous solution by ozone and UV/ozone processes. AB - The decomposition of 2-nitrophenol in aqueous solutions by ozone and UV/ozone processes was found to be technically feasible under adequate experimental conditions. Formation of nitrate ions was observed following the decomposition of 2-nitrophenol by ozone and UV/ ozone processes. Increasing ozone dosage and UV light intensity accelerated the decomposition rate of 2-nitrophenol in an aqueous solution. The species distribution of 2-nitrophenol under various solution conditions plays a significant role in determining decomposition behavior. In most experiments conducted in this study, the decomposition of 2-nitrophenol by ozone and UV/ozone processes was favored to occur in alkaline conditions. The addition of 2-butanol accelerated the rate of gaseous ozone transfer to an aqueous phase by reducing the surface tension of aqueous solution and therefore enhancing the decomposition rate of 2-nitrophenol by ozone and UV/ozone processes. PMID- 17120451 TI - Computational fluid dynamics analysis of the effects of reactor configuration on disinfection efficiency. AB - The efficacy of disinfection processes in water purification systems is governed by several key factors, including reactor hydraulics, disinfectant chemistry, and microbial inactivation kinetics. The objective of this work was to develop a computational fluid dynamics (CFD) model to predict velocity fields, mass transport, chlorine decay, and microbial inactivation in a continuous flow reactor. The CFD model was also used to evaluate disinfection efficiency in alternative reactor designs. The CFD reactor analysis demonstrates that disinfection efficiency is affected by both kinetics and mixing state (i.e., degree of micromixing or segregation). Residence time distributions (RTDs) derived from tracer analysis do not describe intrinsic mixing conditions. The CFD based disinfection models account for reactor mixing patterns by resolution of the reactor velocity field and thus provide a better prediction of microbial inactivation than models that use an RTD. PMID- 17120452 TI - Mechanism of effective nocardioform foam control measures for non-selector activated sludge systems. AB - Solids retention time (SRT), biological scum trapping and recycle, and the dynamic equilibrium between Nocardioform populations in the foam and the mixed liquor are the controlling factors in activated sludge foaming events caused by Nocardioform bacteria. For the operating modes described in this paper, a cured mixed liquor foaming condition (filament counts of approximately 10(5) intersections/g volatile suspended solids) was only achieved when SRT control, selective wasting, and polymer addition were in effect. Solids retention time control, with the SRT remaining below 1.5 days, and selective wasting will cure a severely foaming mixed liquor, but effects will only be observed after 3 or 4 months after implementation. The combined wastage of Nocardioform bacteria from selective wasting and SRT control can ensure long-term foam control to the operation of a pure-oxygen activated sludge system with foam-trapping features. An SRT of 0.3 days will result in the complete washout of Nocardioform bacteria from the activated sludge system, which can then operate at an SRT of 3 days free of Nocardioform. Polymer addition to mixed liquor is only effective for foam control when a large portion of the system biomass exists as a heavy layer of foam above the mixed liquor. PMID- 17120453 TI - Effects of pH, temperature, and water quality on chloride removal with ultra-high lime with aluminum process. AB - The ultra high-lime with aluminum process (UHLA) has the ability to remove sulfate and chloride in addition to other scale-forming materials from recycled cooling water. Laboratory experiments have demonstrated that the UHLA process can achieve high chloride removal from recycled cooling water, and an equilibrium model was developed to describe chemical behavior during chloride removal. This paper describes the influence of pH, temperature, and initial chloride concentration on chloride removal by UHLA and identifies the precipitated solids formed during treatment. The optimum pH for maximum chloride removal efficiency was found to be 12 +/- 0.2. Chloride removal efficiency was higher at a high initial chloride concentration than at a low initial chloride concentration with the chemical doses used. Solids formed during UHLA treatment were identified by x ray diffraction as calcium chloroaluminate, tricalcium hydroxyaluminate, and tetracalcium hydroxyaluminate. This supports the assumption of the equilibrium model that these compounds are present and form a solid solution. PMID- 17120454 TI - Bagasse-assisted bioremediation of ammonia from shrimp farm wastewater. AB - Development of new economically feasible ecofriendly products from agricultural wastes or byproducts for shrimp farm wastewater treatment is the objective of our continued research. Ammonia is a nitrogenous toxicant, which is commonly found in wastewater from shrimp farms. In the present study, we explored the possibility of the use of simply and inexpensively prepared bagasse products so that this abundant crop byproduct could be used to remove ammonia from shrimp farm wastewater. Bagasse, a natural highly fibrous lignocellulosic byproduct of sugarcane, was converted into five different products. Experimental results have shown that ammonia is efficiently removed from wastewater by four bagasse products with a dose of 1 to 6 g/L within 24 hours. The effect of bagasse products on other water quality parameters and growth kinetics of biofilm bacteria onto bagasse fiber have also been studied. Efficacies of products were compared by using statistical analysis. Products developed from bagasse are useful and economical. PMID- 17120455 TI - Municipal landfill leachate treatment for metal removal using water hyacinth in a floating aquatic system. AB - Experiments were carried out to investigate the ability of water hyacinth (Eichhornia crassipes) to remove five heavy metals (cadmium, chromium, copper, nickel, and lead) commonly found in leachate. All experiments were conducted in batch reactors in a greenhouse. It was found that living biomass of water hyacinth was a good accumulator for copper, chromium, and cadmium. The plants accumulated copper, chromium, and cadmium up to 0.96, 0.83, and 0.50%, respectively, of their dry root mass. However, lead and nickel were poorly accumulated in water hyacinth. Also, nonliving biomass of water hyacinth dry roots showed ability to accumulate all metals, except Cr(VI), which was added in anionic form. The highest total metal sorption by nonliving dry water hyacinth roots was found to take place at pH 6.4. The current research demonstrates the potential of using water hyacinth for the treatment of landfill leachate containing heavy metals. PMID- 17120456 TI - Development of a three-stage system for wastewater toxicity monitoring: a design and feasibility study. AB - A three-stage system was developed to automate a batchwise toxicity testing protocol designed for assessing wastewater toxicity to activated sludge. The three-stage system used the luminescent bacterium Shkl. The three stages were cell storage, cell activation, and continuous toxicity testing. Shkl cells were stored in a bioreactor at 4 degrees C when the system was not in use and activated in another bioreactor for use in toxicity tests conducted in a continuous manner. The system could quickly be switched between the "off" and "on" modes, and operation of the system was easy. The stability of the system, in terms of cell density and bioluminescence in the storage and activation bioreactors, and the response of the activated cells to a metal and an organic toxicant were studied. The feasibility of the system design was demonstrated by simulating zinc toxicity episodes in synthetic wastewater. The needs for further modifications and improvements of the system were discussed. PMID- 17120457 TI - Treatment of food industry waste by bench-scale upflow anaerobic sludge blanket anoxic-aerobic system. AB - An upflow anaerobic sludge blanket (UASB)-anoxicaerobic system was used for treatment of tomato and bean processing wastewater. At various hydraulic retention times, ranging from 0.7 to 5 days, excellent removal of chemical oxygen demand (COD), biochemical oxygen demand (BOD), total suspended solids (TSS), ammonia-nitrogen (NH4-N), and total Kjeldahl nitrogen was achieved with final effluent BOD/TSS/NH4N concentrations of less than 15/15/1 mg/L. Biogas yield in the UASB reactor varied from 0.33 to 0.44 m3/kgCODremoved. The kinetics of anaerobic treatment were investigated. The yield coefficient was 0.03 gVSS/gCOD; maximum specific growth rate was 0.24 day(-1); Monod half velocity constant was 135 mgCOD/L; and specific substrate utilization rate was 3.25 gCOD/gVSS x d. Nitrification and denitrification kinetics were studied in batch experiments, and the rates were comparable with those in the continuous flow system. PMID- 17120458 TI - Crystalline hydrous ferric oxide: an adsorbent for chromium(VI)-contaminated industrial wastewater treatment. AB - Synthetic crystalline hydrous ferric oxide (CHFO) (particle size 0.14 to 0.29 mm) has been used systematically for adsorptive chromium(VI) removal from contaminated water. Batch experiments were performed as a function of pH, contact time, solute concentration, and regeneration of adsorbents. Column experiments were performed for breakthrough points in the presence and absence of other ions and treatment of industrial effluent. The optimum pH range was 2.0 to 4.0. The adsorption kinetic data could be described well by both second-order and pseudo first-order models. The isotherm adsorption data at 30 +/- 2 degrees C obeyed the Langmuir model best. The monolayer adsorption capacity was 35.7 mg/g. Chromium(VI)-rich CHFO could be regenerated up to 89 +/- 1% with 2.0 M sodium hydroxide. Regenerated column reuse showed a decrease (10 to 12%) in breakthrough capacity. Finally, the CHFO- (dried at 300 degrees C) packed column was used for the recovery (98.5 +/- 1.0%) of chromium(VI) from contaminated industrial waste effluent of Hindustan Motor Limited (Hooghly, West Bengal, India). PMID- 17120459 TI - Treatment of distillery spent-wash by ozonation and biodegradation: significance of pH reduction and inorganic carbon removal before ozonation. AB - This study is aimed at exploring strategies for mineralization of refractory compounds in distillery effluent by anaerobic biodegradation/ozonation/aerobic biodegradation. Treatment of distillery spent-wash used in this research by anaerobic-aerobic biodegradation resulted in overall COD removal of 70.8%. Ozonation of the anaerobically treated distillery spent-wash was carried out as is (phase I experiments) and after pH reduction and removal of inorganic carbon (phase II experiments). Introduction of the ozonation step resulted in an increase in overall chemical oxygen demand (COD) removal, with the highest COD removals of greater than 95% obtained when an ozone dose of approximately 5.3 mg ozone absorbed/mg initial total organic carbon was used. The COD removal during phase II experiments was slightly superior compared with phase I experiments at similar ozone doses. Moreover, efficiency of ozone absorption from the gas phase into distillery spent-wash aliquots was considerably enhanced during phase II experiments. PMID- 17120460 TI - Recurrence of fecal coliforms and Salmonella species in biosolids following thermophilic anaerobic digestion. AB - The U.S. Environmental Protection Agency (U.S. EPA) Part 503 Biosolids Rule requires the fecal coliform (indicator) or Salmonella species (pathogen) density requirements for Class A biosolids to be met at the last point of plant control (truck-loading facility and/or farm for land application). The three Southern Californian wastewater treatment plants in this study produced biosolids by thermophilic anaerobic digestion and all met the Class A limits for both fecal coliforms and Salmonella sp. in the digester outflow biosolids. At two plants, however, a recurrence of fecal coliforms was observed in postdigestion biosolids, which caused exceedance of the Class A limit for fecal coliforms at the truck loading facility and farm for land application. Comparison of observations at the three plants and further laboratory tests indicated that the recurrence of fecal coliforms can possibly be related to the following combination of factors: (1) incomplete destruction of fecal coliforms during thermophilic anaerobic digestion, (2) contamination of Class A biosolids with fecal coliforms from external sources during postdigestion, (3) a large drop of the postdigestion biosolids temperature to below the maximum for fecal coliform growth, (4) an unknown effect of biosolids dewatering in centrifuges. At Hyperion Treatment Plant (City of Los Angeles, California), fecal coliform recurrence could be prevented by the following: (1) complete conversion to thermophilic operation to exclude contamination by mesophilically digested biosolids and (2) insulation and electrical heat-tracing of postdigestion train for maintaining a high biosolids temperature in postdigestion. PMID- 17120461 TI - Detection of Cryptosporidium parvum oocysts in sediment and biosolids by immunomagnetic separation. AB - A method for the detection of Cryptosporidium parvum oocysts in sediment and wastewater biosolids has been developed using immunomagnetic separation kits that were designed for use with water. This method requires no pretreatment of the sediment or biosolids samples before the commercial kit application. Oocyst recovery efficiencies from sediment and biosolids using the modified Dynal (Lake Success, New York) and Crypto-Scan commercial methods (Immucell Corporation, Portland, Maine) ranged from 20 to 60%. While the sensitivity of the method is dependent on the amount of sediment processed and the equivalent volume examined under the microscope, it was able to detect 0.48 oocysts per gram dry weight sediment. Using this method, Cryptosporidium parvum oocysts were found at levels as high as 97 oocysts/g of primary biosolids and at levels up to 4 oocysts/g in polluted sediment. PMID- 17120462 TI - Compliance with therapy in hypertensive patients. PMID- 17120463 TI - Perioperative myocardial infarction in non-cardiac surgery. Pathophysiology and clinical implications. AB - Advances in surgical and anaesthetic techniques and an aging patient population have resulted in more complex procedures being performed in greater numbers of aged subjects and in patients with a high likelihood of significant cardiovascular disease. Nearly one fourth of non-cardiac surgical procedures (major intra-abdominal, thoracic, vascular, and orthopaedic procedures) performed in persons older than 65 years have been found to be associated with significant perioperative cardiovascular morbidity and mortality. During previous years the main attempt was to define strategies to accurately estimate perioperative cardiovascular risk based either on the characteristics of surgery and on patient characteristics. More recently preventive medical strategies have been proposed. Therefore, the physician has to be aware of the key elements useful to calculate the perioperative cardiovascular risk, and of the medical preventive treatment or further interventions to adopt in patients candidate to surgery. PMID- 17120464 TI - Immunological tolerance and autoimmunity. AB - Immunological tolerance is a complex series of mechanisms that impair the immune system to mount responses against self antigens. Central tolerance occurs when immature lymphocytes encounter self antigens in the primary lymphoid organs, and consequently they die or become unreactive. Peripheral tolerance occurs when mature lymphocytes, escaped from negative selection during ontogeny, encounter self antigens in secondary lymphoid organs and undergo anergy, deletion or suppression. A heterogeneous family of T regulatory cells has recently been identified, which have been found to play an important role in suppressing immune responses against self. Failure or breakdown of immunological tolerance results in autoimmunity and autoimmune diseases. Such events are related to both genetic and environmental factors, the latter being mainly represented by infections. Infectious agents can indeed promote autoimmune responses either by inducing tissue inflammation and therefore an unintended bystander activation of autoreactive T cells, or by promoting T cell responses to microbial epitopes that cross react against self peptides. PMID- 17120465 TI - Subintensive care unit for the elderly: a new model of care for critically ill frail elderly medical patients. AB - OBJECTIVE: An increasing number of elderly patients are admitted to the hospital for critical diseases and the gap between supply and demand of intensive care resources is a growing problem. To meet this challenge, 4 beds in a 24-bed acute care for the elderly (ACE) medical unit were dedicated to a subintensive care unit (SICU). Severely ill elderly medical patients, requiring a higher level of care than provided in ordinary wards, are admitted. The aim of the study was to describe the characteristics of the setting and to discuss its usefulness based on data obtained after the first period of implementation. METHODS: This article describes the development, management, economics and patient characteristics of the SICU. Patient care combines the ACE model with a highly specialised medical care. Patients admitted to the SICU are compared with patients treated in the ordinary ACE unit before the SICU opened. All patients received a multidimensional evaluation, including demographics, main diagnosis, number of chronic somatic diseases, Charlson index, APACHE II score, APACHE-APS subscore, number of currently administered drugs, serum albumin, cognitive status (Mini Mental State Examination), depression (Geriatric Depression Scale) and functional status (basic and instrumental activities of daily living). Ward physicians performed assessment and collection of data. RESULTS: During the first 16 months, 489 patients were admitted, 401 according to the selection criteria (60 +/- years and APACHE II score > or =5 and/or APACHE-APS score > or =3). Mean age was 78.1 years, mean APACHE II score 14.5 (moderate severity) and non-invasive mechanical ventilation was received by 87 (21.7%). The most common diagnoses were respiratory failure, cardiac disease and stroke. Mean length of stay in the SICU was 61.8 h, and 6.0 days in the hospital. Compared with ACE-unit patients admitted during 2002 (n=1380), SICU patients were obviously more seriously ill (APACHE II score 14.5 vs 6.7). When comparing patients of same illness severity (APACHE-APS score > or =3) (n=125), patients treated in the SICU had lower in hospital mortality than those treated in the ordinary ACE ward (12.5 vs 19.2%). Only a few patients (3.5%) were transferred to the intensive care unit as a consequence of increased severity of illness. CONCLUSIONS: The SICU is an innovative method to treat frail elderly patients with more severe conditions. Low hospital mortality compared with that of severe patients in the ACE unit supports the usefulness of this model. It could be implemented in medical units of large hospitals in order to give optimal care and advanced interventions to the frail elderly and to avoid intensive care unit overcrowding. PMID- 17120466 TI - Compliance with therapy in hypertensive patients. AB - OBJECTIVE: This study investigated pharmacological compliance in hypertensive patients recruited during a 24-month period at the Hypertension Clinic of the S. Maria Nuova Hospital. METHODS: The compliance profile scale was developed in order to classify compliance as sufficient, or poor. Compliance was evaluated according to patients' (1) global cardiovascular risk (hypertensive) and (2) complexity of prescribed treatment at enrolment (drug, number of pills). RESULTS: A total of 367 patients were recruited; 82 were excluded owing to changes in therapeutic schedule or medication necessitated by other medical or surgical disease. Compliance was evaluated in 285 patients, 66% were male; the mean age was 72 +/- 12 years. Compliance was evaluated at a series of follow-ups at 3 month intervals and at 36 months of observation. Compliance at 36 months was good or sufficient in 56% of patients, but differed according to hypertensive and drug status. Compliance was better in patients with a higher level vs. lower level of hypertension severity. Specifically, compliance was better in hypertensives with co-morbidity (76%) than in hypertensives without co-morbidity (37%) or hypertensives with risk factors (65%). Moreover, compliance was better in patients with a more complex therapeutic regimen (drug 1 through 3: 47, 59 and 66%). CONCLUSIONS: These data demonstrate a relationship between disease severity and compliance, independent of the prescribed therapeutic regimen. This suggests that the most important factor for therapy adherence is disease awareness and severity, and patients with a higher level of disease are more likely to be compliant. PMID- 17120467 TI - A young woman with oedema. PMID- 17120468 TI - Diffuse acute abdominal pain and palpation of the lower limb pulses: what kind of link? PMID- 17120469 TI - The diagnostic labyrinth. PMID- 17120470 TI - Determining disease severity in severe sepsis and septic shock. PMID- 17120471 TI - Sepsis and its consequences. PMID- 17120472 TI - Cardiac markers in the low-risk chest pain patient. AB - With the arrival of point-of-care cardiac marker determination, emergency physicians may be able to arrive at the diagnosis of cardiac ischemia faster than ever before. However, these tests must be used with care, as a lack of understanding about when and how they should be obtained is important both for good patient care and to avoid medicolegal pitfalls. This report reviews risk stratification of patients who present with chest pain, provides an overview of cardiac markers and literature supporting their use, and concludes with a practice guideline for the utilization of cardiac markers in the emergency department. PMID- 17120473 TI - Prognostic indexes of septic syndrome in the emergency department. AB - OBJECTIVE: Sepsis is a frequent and often lethal condition. Rapid identification and aggressive therapy in the emergency department (ED) are essential for outcome. Several indexes were found to be significantly related to short-term clinical outcome, but only bedside, rapidly available tests are thought to be useful in the ED. To define the prevalence and mortality of patients with severe sepsis presenting to the ED of a tertiary care hospital in Italy, we furthermore investigated the ability of bedside, non-invasive prognostic indexes to identify patients with adverse short-term clinical outcome. METHODS: All patients admitted to the ED with a diagnosis of severe sepsis or septic shock were included. Retrospective data were collected by a dedicated software program using predefined searching criteria including clinical data, vital sign parameters, sepsis-related organ failure assessment (SOFA) score, and blood tests. The relationship between prognostic indexes and 24-h or 28-day mortality was evaluated by multivariate logistic regression analysis. RESULTS: Ninety patients were enrolled from June 2004 to June 2005 (0.2% of all incoming patients to ED and 0.7% of all critical patients). Mean age was 77 +/- 15 years, 54.4% were women. During follow-up (28 days) 46 patients died (51.1%), 21 patients (23.3%) within 24 h. At multivariate analyses, age >80 (odds ratio [OR] 4.10; 95% confidence intervals [CI] 1.39-11.90, p = 0.01), serum lactate >5 mmol/l (OR 3.40; 95% CI 1.21-9.60, p = 0.02) and acute renal failure (OR 18.90; 95% CI 1.80 200, p = 0.02) were independent predictors of 28-day mortality. CONCLUSIONS: Among critical patients admitted to an Italian ED, those with severe sepsis/septic shock represent about 1%, with a very high mortality rate. Bedside non-invasive prognostic indexes are able to identify with high accuracy patients with adverse short-term clinical outcome. PMID- 17120474 TI - A 46-year-old man with difficulty breathing and the sensation of choking. PMID- 17120475 TI - Pleuritic chest pain in a patient who had undergone recent surgical repair of a patent foramen ovale. PMID- 17120476 TI - Human albumin solution for resuscitation and volume expansion in critically ill patients. AB - BACKGROUND: Human albumin solutions are used in a range of medical and surgical problems. Licensed indications are the emergency treatment of shock and other conditions where restoration of blood volume is urgent, burns, and hypoproteinaemia. Human albumin solutions are more expensive than other colloids and crystalloids. OBJECTIVES: To quantify the effect on mortality of human albumin and plasma protein fraction (PPF) administration in the management of critically ill patients. SEARCH STRATEGY: We searched the Cochrane Injuries Group trials register, Cochrane Central Register of Controlled Trials, Medline, Embase and BIDS Index to Scientific and Technical PROCEEDINGS: Reference lists of trials and review articles were checked, and authors of identified trials were contacted. The search was last updated in August 2004. SELECTION CRITERIA: Randomised controlled trials comparing albumin/PPF with no albumin/PPF, or with a crystalloid solution, in critically ill patients with hypovolaemia, burns or hypoalbuminaemia. DATA COLLECTION AND ANALYSIS: We collected data on the participants, albumin solution used, mortality at the end of follow up, and quality of allocation concealment. Analysis was stratified according to patient type. MAIN RESULTS: We found 32 trials meeting the inclusion criteria and reporting death as an outcome. There were 1632 deaths among 8452 trial participants. For hypovolaemia, the relative risk of death following albumin administration was 1.01 (95% confidence interval 0.92-1.10). This estimate was heavily influenced by the results of the SAFE trial, which contributed 91% of the information (based on the weights in the meta-analysis). For burns, the relative risk was 2.40 (1.11-5.19) and for hypoalbuminaemia the relative risk was 1.38 (0.94-2.03). There was no substantial heterogeneity between the trials in the various categories (chi2 = 21.86, df = 25, p = 0.64). The pooled relative risk of death with albumin administration was 1.04 (0.95-1.13). CONCLUSIONS: For patients with hypovolaemia there is no evidence that albumin reduces mortality when compared with cheaper alternatives such as saline. There is no evidence that albumin reduces mortality in critically ill patients with burns and hypoalbuminaemia. The possibility that there may be highly selected populations of critically ill patients in which albumin may be indicated remains open to question. However, in view of the absence of evidence of a mortality benefit from albumin and the increased cost of albumin compared to alternatives such as saline, it would seem reasonable that albumin should only be used within the context of well concealed and adequately powered randomised controlled trial. PLAIN LANGUAGE SUMMARY: There is no evidence that giving human albumin to replace lost blood in critically ill or injured people improves survival when compared to giving saline. Trauma, burns or surgery can cause people to lose large amounts of blood. Fluid replacement, giving fluids intravenously (into a vein), is used to help restore blood volume and hopefully reduce the risk of dying. Blood products (including human albumin), non-blood products or combinations can be used. The review of trials found no evidence that albumin reduces the risk of dying. Albumin is very expensive in which case it may be better to use cheaper alternatives such as saline for fluid resuscitation. PMID- 17120477 TI - Haemolytic uraemic syndrome and thrombotic thrombocytopenic purpura in southern Iran. PMID- 17120478 TI - Considerations on the potential teratogenic effects of antiretroviral therapy. PMID- 17120479 TI - Acute ischaemic stroke treated with combined intra-arterial thrombolysis and intravenous tirofiban despite oral anticoagulant therapy at an international normalised ratio > or = 2.0. PMID- 17120480 TI - A case of acute abdomen for the internist. PMID- 17120481 TI - Whipple's disease in a father-son pair. PMID- 17120482 TI - Re-emerging poliomyelitis--is Australia's surveillance adequate? AB - In the past two years there has been a resurgence of polio with 21 previously polio-free countries importing wild poliovirus. Wild poliovirus importations into polio-free areas will continue to occur until endemic transmission is interrupted globally. Australia's acute flaccid paralysis (AFP) surveillance falls well short of the target of more than 80 per cent of AFP cases having two adequate stool specimens taken at least 24 hours apart within 14 days of onset for poliovirus examination. As most AFP cases are hospitalised, AFP should be immediately notifiable by hospitals to public health units or state or territory public health authorities to ensure appropriate follow up, including stool specimens. PMID- 17120483 TI - Burden and causes of foodborne disease in Australia: Annual report of the OzFoodNet network, 2005. AB - In 2005, OzFoodNet sites recorded 25,779 notifications of seven potentially foodborne diseases, which was 12.5 per cent higher than the mean for the previous five years. Diseases with significant increases in 2005, when compared to historical reports include: Shiga toxin-producing Escherichia coli, shigellosis, haemolytic uraemic syndrome, salmonellosis and campylobacteriosis. The most significant increases were those due to Salmonella (13.1%) and Campylobacter (5.1%) because of the frequency of these infections. Reports of listeriosis were lower than previous years and there were only four materno-foetal infections compared to seven in 2004. Sites reported 624 outbreaks of gastroenteritis and foodborne disease in 2005. One hundred and two of these were foodborne and affected 1,926 persons, hospitalised 187 and caused four deaths. Among foodborne outbreaks, Salmonella Typhimurium was the most common pathogen and restaurants were the most common place where food implicated in outbreaks was prepared. Outbreaks associated with fish, poultry meat, and mixed meat dishes were common. There were several large outbreaks of salmonellosis, including one associated with dips at a Turkish restaurant, one with alfalfa sprouts, and two due to egg based dishes. In addition, there were several multi-state investigations of Salmonella infection during 2005, including one large outbreak of S. Typhimurium 135 implicating poultry meat from retail supermarkets. Sites identified a source of infection for 39 per cent (41/104) of investigations into clusters of salmonellosis. Overall, 97.4 per cent of Salmonella notifications on state and territory surveillance databases recorded complete information about serotype and phage type. This report highlights the considerable burden of disease from food sources in Australia and the need to continue to improve food safety. PMID- 17120484 TI - Communicable Diseases Network Australia annual report, 2005. PMID- 17120485 TI - Annual report: Surveillance of adverse events following immunisation in Australia, 2005. AB - This report summarises Australian passive surveillance data for adverse events following immunisation (AEFI) reported to the Adverse Drug Reactions Advisory Committee for 2005, and describes reporting trends over the six year period 2000 to 2005. There were 839 AEFI records for vaccines received in 2005. This is an annual AEFI reporting rate of 4.1 per 100,000 population, the lowest since 2000 and a 22 per cent decrease compared with 2004 (1,081 records; 5.4 AEFI records per 100,000 population). The decrease was not consistent across age groups. Reporting of AEFI increased for children aged <1 year in 2005 (60.7 versus 50.3 per 100,000 population) and decreased for the 7 to <20 year age group (0.9 versus 8.9 per 100,000 population). Dose-based AEFI reporting rates in 2005 were 11.0 per 100,000 doses of scheduled vaccines for children aged <7 years and 2.0 per 100,000 doses of influenza vaccine for adults aged >18 years. The majority of records described non-serious events while 9 per cent (n=72) described AEFIs defined as serious. There was one report of death in an older person following influenza vaccine and one of non-polio acute flaccid paralysis in an infant, both temporally associated with immunisation. The most frequently reported individual AEFI was injection site reaction in children following a fifth dose of diphtheria tetanus-acellular pertussis vaccine (79 reports per 100,000 doses). The increase in the population-based AEFI reporting rate for children aged <1 year in 2005 coincided with the introduction of national immunisation programs for conjugate pneumococcal vaccine in January 2005 and inactivated poliovirus vaccine in November 2005. The fall in reporting rates for older children and adolescents follows the completion of the national meningococcal C catch-up program in early 2005. The consistently low reporting rate of serious AEFIs demonstrates the high level of safety of vaccines in Australia. PMID- 17120486 TI - Annual report of the Australian National Poliovirus Reference Laboratory 2005. AB - In May 1988 the World Health Assembly adopted a resolution for the global eradication of poliomyelitis. Since then two target dates for eradication (2000 and 2003) have passed and the struggle to eradicate the poliovirus continues. Australia's commitment to the worldwide campaign began in December 1994 with the designation of the National Poliovirus Reference Laboratory at the Victorian Infectious Diseases Reference Laboratory and the initiation of acute flaccid paralysis (AFP) surveillance in March 1995. During 2005 the National Poliovirus Reference Laboratory did not isolate any wild or vaccine derived polioviruses from the 42 samples collected from eighteen cases of acute flaccid paralysis in Australian residents. Three Sabin-like polioviruses were isolated from three cases of acute flaccid paralysis but all were considered incidental isolations by the Polio Expert Committee and not implicated in the disease of the patients. After exceeding the World Health Organization target of one case of AFP per 100,000 children aged less than 15 years in 2004, Australia's non-polio AFP rate in 2005 fell to 0.75 cases per 100,000 children. The high number of wild poliovirus importations reported globally in 2005 into previously polio free countries, highlights the need for a sensitive AFP surveillance system within Australia and for specimens from AFP cases to be forwarded to the National Poliovirus Reference Laboratory. PMID- 17120487 TI - Communicable and vaccine-preventable conditions under surveillance by the APSU: 2005 update. PMID- 17120488 TI - A comparison of data sources for the surveillance of seasonal and pandemic influenza in Victoria. AB - Understanding the characteristics of available influenza or influenza-like illness (ILI) surveillance systems is important for seasonal influenza surveillance and pandemic preparedness. We compared five influenza or ILI data sources in Victoria: notifications of laboratory-confirmed influenza to the Victorian Department of Human Services; hospital emergency presentations and hospital admissions; sentinel general practitioner surveillance; and medical locum service surveillance. Seasonal trends for influenza and ILI activity were similar for all data sources. Community ILI surveillance, operating as sentinel GP, locum service or hospital emergency department surveillance, in conjunction with notification of laboratory-confirmed influenza, would provide adequate inter pandemic surveillance for influenza in Victoria and, by extension, in any Australian jurisdiction. Other surveillance systems would be needed for early pandemic case or cluster detection, while pandemic monitoring would be better achieved by a more automated system. PMID- 17120489 TI - Circulation and antigenic drift in human influenza B viruses in SE Asia and Oceania since 2000. AB - During annual influenza epidemics, influenza B viruses frequently co-circulate with influenza A viruses and in some years, such as 2005, large outbreaks have occurred while in other years, the virus virtually disappears. Since 1987 there have been two lineages of influenza B viruses co-circulating in various countries and causing disease in humans. The proportions of these two lineages vary from year to year and country to country. For example, in 2005, the B/Victoria/2/87 lineage was predominant in New Zealand while in Australia the B/Yamagata/16/88 lineage was more common. Antigenic and genetic analysis has revealed gradual movement in the both lineages. Careful monitoring of the two virus lineages is important, as they are antigenically distinct. This is an important consideration for influenza vaccine formulation decisions, as only one influenza B component is traditionally included in the annual trivalent influenza vaccine. PMID- 17120490 TI - Nosocomial and community transmission of measles virus genotype D8 imported by a returning traveller from Nepal. AB - Measles is uncommon in Australia due to effective national vaccination strategies. In mid-2003, a cluster of nine cases of measles occurred in western Sydney. The index case was a 29-year-old traveller recently returned from Nepal. The case presented to hospital and transmitted the disease to two others in the Emergency Department. Further cases resulted from both community and nosocomial transmission. The median age of cases was 24 years, with three cases in children aged under four years. Only one person had a documented history of measles vaccination, a child who had received one dose of vaccine overseas. One case was a 2-month-old infant whose mother was immune and two cases were hospital staff members. Molecular analysis of measles virus isolates from four cases revealed the same D8 genotype, a strain previously identified in Nepal. Staff vaccination strategies implemented as a result of the outbreak were poorly patronised despite nosocomial transmission. As diseases such as measles become rare it is important to thoroughly investigate any outbreaks, and to maintain a high index of suspicion of measles, particularly in travellers presenting with a rash having returned from measles-endemic areas. Genetic analysis is important in tracing the origins of an outbreak, and to confirm relatedness between cases. The highly infectious nature of measles virus also underscores the need for appropriate infection control in minimising risk of nosocomial transmission. Such policies are of increasing importance with the emergence of novel viruses or the threat of pandemic influenza. PMID- 17120491 TI - A profile of HIV testing in Victoria, 1984 to 2004. AB - HIV testing is an important public health strategy and collection of HIV testing data is a component of overall HIV surveillance activities. This paper describes changes in HIV testing patterns in relation to HIV diagnoses in Victoria between 1984 and 2004. HIV testing and diagnosis data were extracted from surveillance databases maintained at the Burnet Institute. The annual number of HIV tests performed in Victoria increased from 2,879 in 1984, to 193,927 in 2004. Between 1991 and 2004, the male HIV testing rate per 100,000 population increased from 2,754 to 3,710 and the female rate from 2,395 to 4,453. The proportion of HIV tests conducted by private laboratories increased from less than 1 per cent in 1991 to 75 per cent in 2004. The number of HIV diagnoses increased from 140 in 1999 to 233 in 2002 and then fell to 217 in 2004. The HIV diagnosis rate per 100,000 tests increased from 98.9 in 1999 to 137.7 in 2000 then decreased to 111.9 in 2004. The overall rate of HIV diagnosis per 100,000 tests was 291.6 for males and 25.9 for females. Increased testing among males is a good outcome considering the majority of HIV diagnoses in Victoria are among men who have sex with men (MSM). Increased testing among females probably relates to increased antenatal screening. The inability to collect sexual orientation and reason for test data limited interpretations. To provide a better understanding of the impact of testing on the HIV epidemiology, especially among MSM, linked HIV sentinel surveillance has been implemented in Victoria. PMID- 17120492 TI - Campylobacter outbreak due to chicken consumption at an Australian Capital Territory restaurant. AB - Campylobacter is the most common cause of bacterial gastroenteritis in Australia, with 15,008 notifications in 2004. This represents only a small fraction of the total cases of Campylobacter. Despite this, outbreaks are rarely reported. This report describes the investigation of an outbreak of campylobacteriosis following a restaurant meal in the Australian Capital Territory. The outbreak was identified by a general practitioner who notified the Health Protection Service, ACT Health. A retrospective cohort investigation of the 27 work colleagues who attended lunch at the restaurant was conducted. Eleven cases were identified with two culture positive for Campylobacter. An association between eating several dishes containing chicken was identified. This outbreak highlights the important identified risk for Campylobacter infection from commercially prepared chicken. It also demonstrates the important role of clinicians in notifying disease outbreaks. PMID- 17120493 TI - National Vaccine Safety Workshop: Summary and draft recommendations. PMID- 17120494 TI - OzFoodNet quarterly report, 1 April to 30 June 2006. PMID- 17120495 TI - Communicable diseases surveillance: Highlights for 2nd quarter, 2006. PMID- 17120496 TI - Sand fly saliva: effects on host immune response and Leishmania transmission. AB - The feeding success of sand flies (Diptera: Phlebotominae) is linked to the vast array of pharmacological substances in their saliva, which interferes with the host haemostasis and immune response. Modification of feeding site plays also an important role in Leishmania transmission. In naive hosts, co-inoculation of saliva and Leishmania parasites increases the chance of successful transmission. Disease exacerbation seems to be associated with enhanced production of type 2 cytokines and selective inhibition of some macrophage functions including the production of NO and H202. On the other hand, hosts repeatedly exposed to sand fly bites develop anti-saliva immune response that results in a protection against Leishmania infection. This led to a new interesting approach to anti Leishmania vaccine--using salivary components to block parasite transmission. The review is therefore focused on the interactions that run between immunomodulatory molecules in sand fly saliva and host immune response, with the impact on Leishmania infection development. Recent studies revealed that saliva-based vaccine for leishmaniasis might be effective and feasible, however, several questions still require to be solved. The knowledge based on experimental mouse model cannot be fully extrapolated to dogs or humans and due to differences in salivary antigens between sand fly species the protective effect is species specific. On the other hand, the specificity of salivary antigens enables the use of anti-saliva antibodies for monitoring the exposure of hosts to sand fly bites and might be used as a marker of risks for Leishmania transmission in endemic areas. PMID- 17120497 TI - Fish-isolated Naegleria strains and their phylogeny inferred from ITS and SSU rDNA sequences. AB - Effort was made to identify Naegleria strains isolated from organs of fish, using phylogenetic analyses of SSU rDNA and ITS sequences. Eighteen fish-isolated strains studied enlarged substantially the so far available set of Naegleria strains characterized by both molecular markers. The phylogenetic analyses of separate and concatenated SSU rDNA and ITS sequences revealed phylogenetic relationships of strains under study; however, they failed to solve classification of fish-isolated strains into species. The sequence similarity of strain-representatives of Naegleria species as well as data obtained on intragenomic variation of ITS sequences discouraged the authors from the definition of new species. The results of the present study provide evidence of a need to re-evaluate the current practice of setting boundaries between species of the genus Naegleria. Sequences obtained in this study have been deposited in GenBank with accession numbers DQ768714-DQ768743. PMID- 17120498 TI - Prosorhynchus maternus sp. n. (Digenea: Bucephalidae) from the Malabar grouper Epinephelus malabaricus (Perciformes: Serranidae) off New Caledonia. AB - A new species, Prosorhynchus maternus sp. n., is described from the serranid fish Epinephelus malabaricus (Bloch et Schneider) in the waters off New Caledonia. It belongs to a group of Prosorhynchus species from serranids in which the uterus is restricted to the postovarian region. Its distinguishing features include the vitellarium relatively distant from the rhynchus, the cirrus-sac relatively distant from the posterior testis, the distinctly pre-equatorial mouth and several other somatic ratios. New records of Prosorhynchus longisaccatus Durio et Manter, 1968 from Epinephelus cyanopodus (Richardson) and Prosorhynchoides lamprelli Bott et Cribb, 2005 from Caranx papuensis Alleyne et Macleay off New Caledonia are also included. PMID- 17120499 TI - The status of Rhoptrobothrium Shipley et Hornell, 1906 (Cestoda: Tetraphyllidea), with redescription of the type species, R. myliobatidis, and description of three new species from two species of Aetomylaeus (Myliobatiformes: Myliobatidae) from Malaysian Borneo. AB - As part of a metazoan parasite survey of elasmobranchs from Malaysian Borneo, specimens of Rhoptrobothrium Shipley et Hornell, 1906 were collected from the eagle rays Aetomylaeus maculatus (Gray) and Aetomylaeus niehofii (Bloch et Schneider). The type species is redescribed from its type host, and a neotype specimen is designated. In addition, three new species of Rhoptrobothrium are described: R. chongi sp. n., R. gambangi sp. n. and R. limae sp. n. Rhoptrobothrium myliobatidis conspicuously differs from the three new species in its lack of a secondary areola; R. limae is distinguished from R. chongi and R. gambangi based on its greater total length; R. chongi possesses conspicuously stalked remi, while R. gambangi possesses short remi, often folded anteriorly. Rhoptrobothrium is somewhat unusual among tetraphyllideans in its possession of a "metascolex," a character it shares with other taxa in the Thysanocephalinae (i.e., Myzocephalus Shipley et Hornell, 1906, Myzophyllobothrium Shipley et Hornell, 1906 and Thysanocephalum Linton, 1889). The morphology of the "metascolex" of Rhoptrobothrium is investigated and new terminology is suggested to standardise the names given to structures constituting a metascolex. As a result, Rhoptrobothrium is considered to possess cephalic peduncle extensions, termed remi. In Rhoptrobothrium, each remus bears, at its distal end, a primary areola, and, in the case of the three new species, also a secondary areola proximal to the primary areola. Myzocephalus and Myzophyllobothrium are tentatively considered to possess remi; the configuration of the "metascolex" of Thysanocephalum, however, is not considered homologous to the condition in the other three genera currently placed in the Thysanocephalinae. PMID- 17120500 TI - A new trematode species Neoplagioporus kajika sp. n. (Digenea: Opecoelidae), parasitic in the Japanese fluvial sculpin, Cottus pollux (Osteichthyes: Scorpaeniformes: Cottidae), from Japan. AB - The adult morphology is described and illustrated of Neoplagioporus kajika sp. n. (Digenea: Opecoelidae) found in the Japanese fluvial sculpin Cottus pollux Gunther (Osteichthyes: Scorpaeniformes: Cottidae) collected in the Naka River at Terase Bridge, Narutake, Nakagawa Town, Fukuoka Prefecture, Kyushu, Japan. This new species is characterized by that the body shape is oval, that the intestinal caeca end posteriorly at the middle level of the testicular region, that the ovary is trilobed, and that the vitelline follicles are distributed between the pharyngeal level and usually the posterior end of body and fill up the lateral fields of body. The new species is different from three hitherto known Neoplagioporus species, N. zacconis (Yamaguti, 1934) Shimazu, 1990 (type species), N. ayu (Takahashi, 1928) Shimazu, 1990, and N. elongatus (Goto et Ozaki, 1930) Shimazu, 1990, in a combination of these characteristics. The new species is considered mainly infective to C. pollux in the river. PMID- 17120501 TI - Comparative description of males of two species of Achtheres von Nordmann, 1832 (Copepoda: Siphonostomatoida: Lernaeopodidae) infecting zander and European perch. AB - Achtheres percarum von Nordmann, 1832 and Achtheres sandrae Gadd, 1901 (Lernaeopodidae) are common parasitic copepods infecting fishes in Eurasia. The former is specific to perch, Perca fluviatilis L., while the latter, to zander, Sander lucioperca (L.). Until recently these copepods have been regarded a single species. The present study was intended to analyse details of male morphology and provide their complete descriptions with differential diagnosis. Males of A. percarum and A. sandrae were collected from perch and zander at Lake Dabie (north western Poland). The males of A. sandrae are larger than those of A. percarum. They also differ in proportions of the first antenna, mandibular denticulation, structure of the first maxilla, and the armament of caudal ramus. The reported differences in male morphology constitute a conclusive confirmation of the separate identity of the two species. PMID- 17120502 TI - Two new species of Apoloniinae (Acari: Trombiculoidea: Leeuwenhoekiidae) from African small mammals, with a key to the species of the world. AB - The genus Liuella Wang et Bai, 1992 is transferred from Trombiculidae to Apoloniinae and a new species Liuella monosetosa sp. n. described from specimens taken in Morocco from the hosts Gerbillus sp. and Meriones libycus Lichtenstein. A new species of Straelensia Vercammen-Grandjean et Kolebinova, 1968, Straelensia variocula sp. n. is described from specimens taken in Morocco and Burkina Faso from the hosts Genetta thierryi Matschie, Gerbillus sp., M. libycus and Elephantulus rozeti (Duvernoy). A list of species, hosts and localities and a key to the species of the Apoloniinae of the world are presented. PMID- 17120503 TI - Some camallanid nematodes from marine perciform fishes off New Caledonia. AB - Two new, one known and three unidentified species of the nematode family Camallanidae are reported and described from the intestines of marine perciform fishes off the southwestern coast of New Caledonia, South Pacific: Camallanus carangis Olsen, 1952 from the forked-tailed threadfin bream Nemipterus furcosus (Nemipteridae), the yellowstriped goatfish Upeneus vittatus and the whitesaddle goatfish Parupeneus ciliatus (both Mullidae) (new host records); Procamallanus (Spirocamallanus) variolae sp. n. from the white-edged lyretail Variola albimarginata (type host) and the blacktip grouper Epinephelus fasciatus (both Serranidae); Procamallanus (Spirocamallanus) longus sp. n. from the twotone tang Zebrasoma scopas (Acanthuridae); Procamallanus (Spirocamallanus) sp. 1 (female tail with 2 terminal spikes on a digit-like projection) from the speckled sand perch Parapercis hexophtalma (Pinguipedidae); Procamallanus (Spirocamallanus) sp. 2 (female tail with 1 spike on a digit-like projection) from the drab emperor Lethrinus ravus (Lethrinidae) and Procamallanus (Spirocamallanus) sp. 3 (female tail with a smooth digit-like protrusion) from the two-lined monocle bream Scolopsis bilineata (Nemipteridae). Camallanus paracarangis Velasquez, 1980 is synonymized with C. carangis. Several additional species of Camallanus from marine fish of the Indo-Pacific region may be synonymous with C. carangis as it has a poorly sclerotized left spicule and 3 small caudal projections on the tail of young (i.e., non-gravid) females. The fourth-stage larva of C. carangis is described for the first time. P. (S.) variolae differs from most similar species of this region mainly in the position (i.e., at level or posterior to the nerve ring) and shape of deirids. P. (S.) longus differs from the similar P. (S.) chaimha mainly in a different arrangement of postanal papillae, shape of the female tail, much longer right spicule (429 microm) and longer body of gravid females (38-55 mm). All Camallanus and Procamallanus (Spirocamallanus) spp. reported here represent the first records of camallanids from marine fishes in New Caledonian waters. PMID- 17120504 TI - Systematic status of Rhabdochona leucaspii Kritscher, 1979 (Nematoda: Rhabdochonidae). AB - Reexamination of the male holotype of the rhabdochonid nematode Rhabdochona leucaspii Kritscher, 1979, originally described from the intestine of the cyprinid fish Leucaspius prosperi [= Ladigesocypris ghigii (Gianferrari)] from Rhodes Island, Greece, confirmed its morphological and biometrical identity with Rhabdochona denudata (Dujardin, 1845), a common and widespread parasite of cyprinids and some other fishes in palaearctic Eurasia. Consequently, R. leucaspii is considered a junior synonym of R. denudata. PMID- 17120505 TI - Awareness about the persons with disability act among leprosy patients and other disabled persons. AB - To assess the level of awareness about the different provisions of the Persons with Disability Act (PWD Act) among leprosy patients and other disabled, 233 disabled persons from the self-help groups formed by Vadathorasalur Leprosy Control Unit have been interviewed using a structured interview checklist. The results show that 74.7% of the respondents were aware that identity cards are available for the disabled, 56.2% were aware of the free education benefit to the disabled, as low as 35.6% were aware of the scholarships, 33% knew about the employment reservations, 24.9% heard about the housing scheme of the government for the disabled, but 24.5% only knew about law against discrimination, 31.8% came in contact with institutions for the severely disabled and only 16% were aware of the unemployment allowance to the disabled. The level of awareness is low among women with regard to all components of the Act. It was found that students studying up to secondary level were not aware of the availability of scholarships and free education, which needs to be seriously looked into, especially by educational institutions. The level of formal education played a significant role in increasing awareness about the Act among literates. The knowledge is low among persons of all occupations. The study showed that there is a great need for an educational intervention programme to publicize the provisions of the Act among the disabled and their families. PMID- 17120506 TI - Profile of new cases of childhood leprosy in a hospital setting. AB - A hospital-based prospective study was carried out to assess the frequency of occurrence of leprosy in childhood. Out of 800 patients registered for leprosy, 67 (8.4%) were children aged 4-14 years. The male-to-female ratio was 2.5:1. Family history of leprosy was found in 14.9% of cases. The commonest type of leprosy was BT leprosy (35.8%), followed by BB leprosy (25.4%) and BL leprosy (19.4%). More than half of the patients had more than one lesion. Nerve involvement was noted in 70.1% of cases. Slit-skin smear was positive in 46.3% of cases. Out of 67 children, PB and MB regimens were given to 29 and 38 respectively. PMID- 17120507 TI - Role of target groups in integrated leprosy programmes. AB - The introduction of integrated leprosy services into the primary health care set up has taken away active case-detection in the community and is replaced by passive reporting by the suspected, afflicted individuals. This can only be made operative effectively with intensive IEC activities in the community. A research study involving school-children (219,000) in leprosy work achieved spectacular success in new case-detection, effective monitoring, completion of MDT and coverage of a large number of individuals (750,000). The results evaluated on a representative sample of 20,000 school students (pre- and post-test), showed over 90% success in creating awareness about the cause of the disease, its symptoms, curability by fixed duration MDT and better attitudes and perceptions of the community towards leprosy-affected individuals. It is emphasised that, in view of the experience gained from the study, other more cohesive and disciplined target groups, such as scouts and guides, NCC cadets, NSS volunteers, should be identified for leprosy work throughout the country in a planned and coordinated manner in order to implement and sustain leprosy eradication activities in the near-elimination and post-elimination phases. PMID- 17120508 TI - Inter-state variations in integration of leprosy services into general health system in low/ moderately endemic states of India. AB - The objective of the study was to analyse inter-state variations in integration of leprosy services into the general health system, covering broad categories of structure integration, training of health functionaries, availability of MDT services and record maintenance, in 24 low/moderately endemic states. Multi-stage random sampling technique was used to select 9 states, 86 health facilities (including district hospitals, community health centres, primary health centres) and 108 sub-centres. Information from each level was collected on a pre-tested form by officers of three leprosy institutions of the Government of India. The results showed wide inter-state variations on each aspect. Redeployment of vertical staff was complete (100%) in Tamil Nadu and Tripura. Assam reported a higher level of training (97%) of medical officers in leprosy. Training of health supervisors and multipurpose workers was better than that of medical officers in most of the states. Tripura reported negligible training of all the health functionaries because of specific local problems. In Assam, Maharashtra and Sikkim, all the urban and rural health facilities were providing MDT. Three months' stock of all types of MDT blister packs was available only in one health facility in Andhra Pradesh and in Goa. Assam and Haryana had lower availability of MDT stocks. In Assam and Maharashtra, medical officers in all health facilities were diagnosing and treating leprosy cases, as compared with Himachal Pradesh where the value was 30%. Involvement of sub-centres in MDT delivery was more at 92% and 100% in Tamil Nadu and Maharashtra respectively in comparison to none in Himachal Pradesh and Tripura. Use of the Simplified Information System (SIS) 2002 guidelines and formats was universal. However, lower involvement of GHS staff in recording and reporting was noted in Assam (0%), Andhra Pradesh (10% and 30%). The study emphasized the need for further tailor-made follow-up studies to suit local problems. PMID- 17120509 TI - Primary neuritic leprosy: a reappraisal at a tertiary care hospital. AB - AIM: To study the clinico-epidemiological profile of primary neuritic leprosy. METHODS: Retrospective analysis of the leprosy records of clinically diagnosed patients of PNL who attended the Leprosy Clinic from 2000 to 2004 was carried out for details of presenting complaints, age and sex distribution, duration, number of nerves involved and pattern of nerve enlargement, BI (skin), nerve abscess, and deformities. RESULTS: There were 32 (4.6%) patients of PNL out of a total of 686. Majority (56.2%) had complaints for less than a year. There were 29 (90.6%) males, and 3 (9.4%) females, with 15-30 years as the commonest age-group (65.6%) involved. Paraesthesia and numbness were the presenting complaints in 20 (62.5%), and motor deficit (paresis) in 11 (34.4%); deformities (claw hand, foot drop, trophic changes) were seen in 16 (50%) cases. Polyneuritic pattern was noted in 21 (63.56%) patients and mononeuritic in 11 (16.5%) with ulnar nerve as the most commonly enlarged nerve (63.6%). Nerve abscess was noted in 4 (12.5%) cases. Slit skin smear was positive in 2 (6.2%) cases only. DISCUSSION: PNL continues to be common in India. Sensory complaints are early and more common. The disease is more common in males. Polyneuritic pattern was predominant, and the ulnar nerve was the most commonly involved nerve. Majority of the cases belong to the tuberculoid spectrum. Early diagnosis depends on complete neurological examination in order to reduce the sequelae of the disease. PMID- 17120510 TI - Affection of immune zones in leprosy: a clinico-epidemiological study. AB - The study was undertaken to evaluate the prevalence of involvement of immune zones in leprosy and to assess the clinico-epidemiological characteristics of the disease in patients presenting with immune zone involvement. 200 leprosy cases were included in this study and detailed history, clinical examination, slit-skin smears and skin biopsies were carried out on all patients. Those cases presenting with immune zone involvement were further evaluated for clinical and epidemiological characteristics of disease process. Immune zone involvement was detected in 7% of cases with male preponderance in the study. Majority of patients (85.5%) had borderline tuberculoid leprosy and midline of back was the commonest site of involvement (50% of cases). Morphologically, macular lesions were the commonest presentation (85.68%) of immune zone involvement. The study heightens the clinical awareness of the possibility of occurrence of leprous lesions on uncommon and unusual sites, which should be termed as relatively immune, rather than absolutely immune zones of leprosy. PMID- 17120511 TI - Dynamic procedures for claw-finger correction--dilemma of balancing tension in transferred tendon slips. AB - The critical step in dynamic claw-finger correction procedures is adjustment of tension on the tendon slips which are being sutured at the new insertion sites to correct finger-clawing. Several methods to balance and adjust the tension have been described ever since these procedures have been in use. Ultimately it is the experience of the operating surgeon that helps to decide as to the tension that is to be kept on each slip so that maximum deformity correction is obtained without compromising the functional capabilities of the hand. An attempt has been made to describe this "experience" in words so that the surgeons who perform these corrective surgical procedures for the first time have some criteria to guide them. PMID- 17120512 TI - Lepromatous leprosy with bilateral facial nerve palsy and hyperthyroidism. AB - Bilateral lagophthalmos secondary to facial nerve is extremely uncommon. Further, the aetiology in most of these cases is of central origin unlike the peripheral involvement in leprosy. A patient of lepromatous leprosy (LL) may be euthyroid or hypothyroid on account of leprous involvement of the thyroid gland. A case of LL with bilateral lagophthalmos and hyperthyroidism is reported. PMID- 17120513 TI - Tattooing in leprosy: the myth continues! PMID- 17120514 TI - Global forum on elimination of leprosy as a public health problem. PMID- 17120515 TI - Externalizing problems through art and writing: experiences of process and helpfulness. AB - Externalization of problems as a component of narrative therapy has been well defined by such authors as Epston and White, and Freedman and Combs. This study reflects the voices and experiences of 17 participants who engaged in an innovative externalization exercise combining sculpture and journaling over a period of 4 weeks. In an attempt to better understand the experience of the participants, the principal investigator also engaged in the exercise. Findings indicated that the intervention helped participants express emotions, increased their awareness of personal resources and agency, helped separate problems from self decreased symptoms and problem behaviors, and fostered a sense of empowerment. This study reveals the potential usefulness of physically externalizing problems and interacting with them deliberately over time. Such interventions may be useful components of narrative therapy or augmentations to brief therapy. They may help reach populations who have limited access to therapy services or who are reluctant to present for therapy. PMID- 17120516 TI - The effect of proxy voice intervention on couple softening in the context of enactments. AB - In this study we evaluated the effectiveness of proxy voice (therapist acting as client's "voice") intervention, embedded within couple enactments, on client perceived softening. The primary research question was whether use of proxy voice would be more likely to bring about softening, or if its use was counterintuitive to enactment conceptualization and would elicit struggle behavior (e.g., withdrawal or negativity). Results indicate that proxy voice has a significant, positive association with softening and is inversely related to withdrawal or negativity. Preliminary findings suggest that proxy voice intervention embedded within a fluid, carefully delineated, and discriminating model of enactments effectively facilitates essential elements of couple interaction (expression of primary affect and articulation of self-concept and attachment threats) while promoting self-reliant couple interaction and increased softening. PMID- 17120517 TI - Factor analysis of the personal authority in the family system questionnaire. AB - The factor structure of the Personal Authority in the Family System Questionnaire (PAFS-Q) has been used in numerous studies, but the factor structure has not been examined since its development in 1984. This study examined the factor structure of the PAFS-Q. Findings suggest a six-factor solution with important differences from those reported when the PAFS-Q was developed. The main differences between this study and the original factor analysis are that this study found separate Mother and Father Intimacy factors instead of a single Intergenerational Intimacy factor and the original Spousal Fusion and Spousal Intimacy factors were not separate factors in the current study. Implications and future directions for research are also discussed. PMID- 17120518 TI - The modified cognitive constructions coding system: reliability and validity assessments. AB - The cognitive constructions coding system (CCCS) was designed for coding clients' expressed problem constructions on four dimensions: intrapersonal-interpersonal, internal-external, responsible-not responsible, and linear-circular. This study introduces, and examines the reliability and validity of a modified version of the CCCS-a version that involves coding clients' problem constructions at the level of speech turn. Results indicate that the modified CCCS generated reliable ratings of clients' problem constructions in 15 sessions of family therapy. Validity was established by comparing 71 subjects' own ratings of over 250 self generated problem constructions to the ratings of trained CCCS coders. The modified CCCS allows for identifying moment-to-moment changes in clients' problem constructions and, ultimately, examining the impact of therapist behaviors on such changes. PMID- 17120519 TI - Directive and nondirective spousal support: differential effects? AB - In this study we investigated whether the following factors influenced married partners' reactions to spousal support: type of support (directive vs. nondirective); individual differences (gender, depressive symptoms); and dimension along which support was evaluated (level of helpfulness vs. valence of emotional impact). Nondirective support was consistently rated more favorably on the dimension of partner's impact than on the dimension of partner's helpfulness, and this effect was exaggerated for those self-reporting a high level of depressive symptoms. Nonetheless, directive support was provided more frequently than was nondirective support in this sample. Gender differences and complications introduced by partner dysphoria suggest some situations in which directive support may be particularly difficult to provide effectively. Implications for couple therapy are discussed. PMID- 17120520 TI - Domestic violence in veterans with posttraumatic stress disorder who seek couples therapy. AB - Domestic violence rates among veterans with posttraumatic stress disorder (PTSD) are higher than those of the general population. Individuals who have been diagnosed with PTSD who seek couples therapy with their partners constitute an understudied population. Self-report measures of domestic violence, relationship satisfaction, and intimacy were administered at intake to 179 couples seeking relationship therapy at a Veterans Affairs clinic. Couples in which the veteran was diagnosed with combat-related PTSD were compared with two other groups based on the veteran's primary diagnosis (depression, adjustment disorder/V-code). Both the PTSD- and depression-diagnosed veterans perpetrated more violence than did those with adjustment/V-code diagnoses. Domestic violence rates among depressed and PTSD-diagnosed veterans were much higher than those found in previous research. Implications for assessment and treatment are discussed. PMID- 17120521 TI - Essential components of curricula for preparing therapists to work effectively with lesbian, gay, and bisexual clients: a Delphi study. AB - The purpose of this study was to determine the knowledge, experiences, and values that therapists should possess to provide quality therapy services to lesbian, gay, and bisexual (LGB) persons and their families. Using the Delphi method, consensus from a panel of experts on LGB topics was obtained on the components that training curricula should ideally include to prepare therapists adequately to work with this population. Panelists provide suggestions for important therapist values and qualities, key theoretical orientations, common issues of LGB clients and their support persons, key components of assessment and interventions, ethical and legal issues, and recommended materials. The panelists stress self-of-the-therapist work and development of a systemic perspective as critical aspects of training. The role of sexuality was deemphasized. Recommendations to training programs based on these findings are provided. PMID- 17120522 TI - Juvenile anorexia nervosa: family therapy's natural niche. AB - Juvenile Anorexia Nervosa (AN) is a severe problem both in terms of presenting symptomatology and its tendency toward chronicity. Researchers have consistently shown that family-based approaches are superior to individual approaches for the treatment of juvenile AN. This article addresses the capacity deficit of trained family therapists to treat this disease. The author reviews the effectiveness of Structural Family Therapy as a treatment of juvenile AN and the essential concepts and skills required by the family therapist to treat this disorder. The concepts of therapeutic crisis induction, enactment, and therapeutic intensity are discussed in detail. Recommendations are made for future research. PMID- 17120523 TI - The implementation and the cultural adjustment of functional family therapy in a Dutch psychiatric day-treatment center. AB - Because of the increasing severity of adolescent problem behavior, evidence-based practices are becoming of interest as an alternative to traditional treatment with the behavior problems of adolescents in juvenile justice settings. Despite interest in evidence-based practices, questions exist regarding whether or not evidence-based intervention models can be successfully transported to cultures other than those in which they were developed. This article describes the transportation process of an American evidence-based family therapy (Functional Family Therapy [FFT]) into the service delivery system of a psychiatric day treatment center for juvenile delinquents in Amsterdam. The characteristics of FFT that make it cross-culturally sensitive are discussed. Results from the changes in service delivery suggest FFT can be successfully implemented in international settings with adjustments to make the model fit the culture(s) of The Netherlands without changing the model of FFT itself. PMID- 17120524 TI - Phytoremediation of selenium using subsurface-flow constructed wetland. AB - The potential of two plant species, Phragmites australis (common reed) and Typha latifolia (cattail), in the phytoremediation process of selenium (Se) was studied in subsurface-flow constructed wetland (SSF). Se was supplemented continuously at a concentration of 100 microg Se L(-1) in the inlet of the cultivation beds of the SSF. Water samples collected from the outlet of the Phragmites bed after 1, 3, 6, 9, and 12 wk of treatments showed that Se content was under detectable limits. Water samples collected from the Typha bed at the same five periods showed that Se concentrations in the outlet were 55, 47, 65, 76, and 25 microg/L, respectively. The results of bioaccumulation in the biomass of both species after 12 wk of treatment indicated that Typha plants accumulated Se mainly in fine roots. Phragmites accumulated Se mainly in leaves and rhizomes, and moderate levels were found in stems and fine organic materials. The results indicate that common reed is a very good species for Se phytoextraction and phytostabilization (immobilization) and that cattail is only a phytostabilization species. The use of common reed and cattail for Se phytoremediation in a SSF system and in constructed wetland models are discussed. PMID- 17120525 TI - Potential for phytoremediation of polychlorinated biphenyl-(PCB-)contaminated soil. AB - Weathered soils contaminated with commercial-grade Aroclor 1260 from three sites in Canada were used to investigate the polychlorinated biphenyl (PCB) phytoextraction potential of nine plant species (Festuca arundinacea, Glycine max, Medicago sativa, Phalaris arundinacea, Lolium multiflorum, Carex normalis, and three varieties of Cucurbita pepo ssp. pepo) under controlled greenhouse conditions. The soils used varied in PCB concentration (90-4200 microg/g) and total organic content (0.06-2.02%). Greenhouse experiments controlled for PCB volatilization through the use of a vented enclosure and by isolating the contaminated soils with parafilm. After 8 wks, PCB concentrations of 47-6700 microg/g were observed in root tissues. Although PCB concentrations in shoot tissues were lower (< 1-470 microg/g), the absolute amounts of PCBs observed in shoot tissue were significant (1.7-290 microg) once shoot biomass was accounted for. Congener signatures indicated that tetra- to hexa-chlorobiphenyls contributed the largest proportions to shoot tissues, but hepta-to nona chorobiphenyls were also present in measurable amounts. Overall, the results indicate that varieties of C. pepo were more effective at extracting PCBs from soil than other plants screened The evidence suggests that this was mainly due to root uptake of PCBs and tranlocation to the shoots, rather than volatilization of PCBs from soil. All plants screened showed signs of stress in the most highly contaminated soil (4200 microg/g), but not in the two lower PCB contaminated soils (250 and 90 microg/g, respectively). No detectable decreases in soil PCB concentrations were observed in these short-term greenhouse experiments, but the results suggest that this may be achievable through multiple plantings. PMID- 17120526 TI - Phytoremediation of two types of sediment contaminated with Zn by Schoenoplectus americanus. AB - The effect of different sediments on growth, Zn uptake, Zn plant distribution, and morphometric variables of Schoenoplectus americanus were investigated under controlled conditions. Two types of sediments were assayed: from a large natural levee (LS) and alluvial sediments (AS), the former with lower organic matter (OM) and nutrients content than AS, without and with added Zn (2500 microg Zn/g air dry sediment). Zinc partition in sediment was determined. Increases in water conductivity and Zn concentrations in water and sediments were observed in artificially contaminated treatments. Plants showed a lower above ground growth rate, height, and width of shoots, and a higher Zn concentration in shoots and rhizomes. In the contaminated treatments, AS treatment showed lower Zn concentration in water and higher Zn concentration in sediments (total, exchangeable, and OM fractions) than LS treatment, due to Zn displacement from floodwater to sediments. The presence of a high level of OM and nutrients also increased aboveground biomass growth, whereas it decreased Zn concentration in shoots. Although the translocation factor increased with Zn addition, it was lower in AS treatment Sediments of AS treatments are a suitable environment for growth of S. americanus, which partially compensates the toxic effects of Zn. Our results provide an encouraging basis for planning larger scale experiments to test the role of OM and nutrients in improving phytoremediation. PMID- 17120527 TI - Stimulating natural defenses in poplar clones (OP-367) increases plant metabolism of carbon tetrachloride. AB - Groundwater contamination by carbon tetrachloride (CCl4) presents a health risk as a potential carcinogen and pollutant that is capable of depleting the ozone layer. Although use of poplar trees in a phytoremediation capacity has proven to be cost effective for cleaning contaminated sites, minimizing leaf emission of volatile contaminants remains a pressing issue. We hypothesized that recently fixed carbon plays a key role in CCl4 metabolism in planta yielding nonvolatile trichloroacetic acid (TCA) and that the extent of this metabolism can be altered by heightening plant defenses. Labeling intact leaves with (11)CO2 (t 1/2 20.4 m) can test this hypothesis, because the extremely short half-life of the tracer reflects only those processes involving recently fixed carbon. Using radio-HPLC analysis, we observed [(11)C]TCA from leaf extract from poplar clones (OP-367) whose roots were exposed to a saturated solution of CCl4 (520 ppm). Autoradiography of [(11)C]photosynthate showed increased leaf export and partitioning to the apex within 24 h of CCl4 exposure, suggesting that changes in plant metabolism and partitioning of recently fixed carbon occur rapidly. Additionally, leaf CCl4 emissions were highest in the morning, when carbon pools are low, suggesting a link between contaminant metabolism and leaf carbon utilization. Further, treatment with methyljasmonate, a plant hormone implicated in defense signal transduction, reduced leaf CCl4 emissions two-fold due to the increased formation of TCA. PMID- 17120528 TI - Laboratory study of heavy metal phytoremediation by three wetland macrophytes. AB - Detention ponds and constructed wetlands have proven to be effective in reducing peak stormwater runoff volume and flow, and recent interest has extended to utilizing them to improve stormwater runoff quality. A review of stormwater runoff studies indicated that lead, zinc, copper, cadmium, phosphorus, and chloride are contaminants of primary concern. In laboratory settings, the uptake of contaminants by three wetland plant species, Glyceria grandis, Scirpus validus, and Spartina pectinata, was examined and removal rates from nutrient solutions inflow and nonflow reactors were measured. The removal rates varied by plant species and target contaminant, and no one species was the best accumulator of all six contaminants. Belowground tissues of all three species accumulated higher concentrations of the four heavy metals and aboveground tissues accumulated higher concentrations of phosphorus and chloride. Plants grown in flow reactors showed significantly higher accumulation rates than those grown in nonflow reactors. Also, plants grown hydroponically accumulated higher concentrations of the six target contaminants than those grown in sand reactors. However, those grown in sand had a much greater increase of biomass and removed a greater mass of the six target contaminants. Removal rates measured in these experiments can be used to design detention ponds to maximize stormwater remediation. PMID- 17120529 TI - Growth conditions impact 2,2-bis(p-chlorophenyl)-1,1-dichloroethylene (p,p'-DDE) accumulation by Cucurbita pepo. AB - Laboratory experiments were conducted to study the effects of soil moisture content, planting density, plant age, and the growth of multiple generations on the bioconcentration of weathered p,p'-DDE by the plant Cucurbita pepo. As soil moisture content increased from 7.4% to 29.9% (by weight), rates of contaminant accumulation by plant roots were increased by more than a factor of 2. Higher planting density also led to higher uptake, as the root bioconcentration factor (BCF, dry-weight ratio of contaminant concentration in the tissue to that in the soil) increased by 15-fold as the number of plants per pot was raised from 1 to 3. Concentrations of the compound in plant roots were inversely related to plant age, with root BCF declining by approximately a factor of 3 as plants aged from 14 to 28 d. Finally, no change in the bioavailability of the compound was observed in successive generations of plants grown in the same contaminated soil. The results suggest that phytoremediation is influenced by a number of factors and that the cleanup of contaminated soil can be enhanced by an understanding of environmental and other conditions affecting plant growth and bioconcentration. PMID- 17120530 TI - Formation of DHP-derived DNA adducts in vivo from dietary supplements and chinese herbal plant extracts containing carcinogenic pyrrolizidine alkaloids. AB - We recently determined that the metabolism of a series of tumorigenic pyrrolizidine alkaloids resulted in the formation of a set of 6,7-dihydro-7 hydroxy-1-hydroxymethyl-5H-pyrrolizine (DHP)-derived DNA adducts. These DHP derived DNA adducts have been proposed as potential biomarkers of pyrrolizidine alkaloid tumorigenicity, as well as pyrrolizidine alkaloid exposure. In this paper, we report that DHP-derived DNA adducts are formed in the liver of female F344 rats, gavaged with three dietary supplements (comfrey root extract, comfrey compound oil, and coltsfoot root extract), or an extract of a Chinese herbal plant, flos farfara (Kuan Tong Hua). PMID- 17120531 TI - Plasma lipid peroxidation and erythrocyte antioxidant enzymes status in workers exposed to cadmium. AB - BACKGROUND: Cadmium (Cd) is a corrosion-resistant metal, used extensively for electroplating in the automobile, electronic and aerospace industry. Only a few studies are available regarding Cd-induced oxidative stress in animals, but no reports are available regarding the effects of Cd on oxidative stress during occupational exposure. OBJECTIVE: The present study was carried out to determine the plasma lipid peroxidation and erythrocyte antioxidant enzymes status in workers exposed to Cd during electroplating. METHODS: 50 subjects exposed to Cd during electroplating formed the study group. An equal number of age-sex matched subjects, working in the administrative section, formed the control group. Urinary Cd levels were determined using the flameless atomic absorption spectrophotometer. Plasma lipid peroxidation and erythrocyte antioxidant enzymes were determined using spectrophotometric methods. RESULTS: A significant increase of plasma lipid peroxidation and a significant decrease of superoxide dismutase and glutathione peroxidase levels were noted in the study group compared with the control group. The level of plasma lipid peroxidation was positively and erythrocyte antioxidant enzymes were negatively and significantly associated with Cd levels in urine. Multiple regression analysis assessed the oxidative stress associated with Cd and other lifestyle confounding factors, such as age, body mass index, the consumption of vegetables, coffee, tea, smoking and alcohol. Analysis showed that the lifestyle confounding factors viz; smoking, body mass index and urinary Cd levels > 5 microg/g of creatinine, were significantly associated with oxidative stress. CONCLUSION: The results of the present study suggest that increased plasma lipid peroxidation and decreased superoxide dismutase levels could be used as biomarkers of oxidative stress in cadmium exposed workers. PMID- 17120532 TI - Effect of sodium azide on the metabolic activity of cultured fetal cells. AB - Sodium azide is a highly toxic substance. However, the mechanism of its toxicity has not been fully established. In the present study, we attempted to investigate the toxicity of sodium azide in various cultured fetal cells, using changes in cellular respiration as an indicator of metabolic inhibition to elucidate tissue specificity. The human fetal cell lines used in this study included myocardial cells, nerve cells, fibroblasts, hepatocytes and renal tubular epithelial cells. The cells were seeded in wells at a density of 2 x 10(6)cells/2mL, sodium azide was added at a concentration of 0.01 ng/mL to 10 microg/mL, and the respiration of each type of cell was measured 1 h later using a dissolved oxygen meter. The concentration at which sodium azide inhibited metabolic activity was lower in the nerve and myocardial cells than in the fibroblasts, hepatocytes and renal tubular epithelial cells. These findings may serve to clarify the dynamic mechanisms of sodium azide toxicity in vivo. PMID- 17120533 TI - The effect of Mentha spicata Labiatae on uterine tissue in rats. AB - The plant Mentha spicata, or peppermint, is commonly used in the treatment of loss of appetite, common cold, bronchitis, sinusitis, fever, nausea and vomiting, and indigestion as a herbal agent. In this study, we aimed to investigate the biochemical and histological effects of M. spicata Labiatae, growing on the Anamas high plateau of Yenisarbademli town, on rat uterine tissue. Twenty female Wistar albino rats weighing 160+/-10 g were used for this study. Rats were divided into two groups of ten animals: group I received no herbal tea (control group) and group II received 20 g/L M. spicata tea. Control group rats were given commercial drinking water (Hayat DANONESA water). Herbal tea was prepared daily and provided at all times to the rats over 30 days as drinking water. Plasma malondialdehyde (MDA) levels were determined. In addition, uterine tissues were submitted for histopathologic examination. MDA levels were increased in group II when compared with the control group. The difference between group II and the control group was statistically significant (P<0.01). In the M. spicata Labiatae treated group, histopathological changes like apoptosis and diffuse eosinophil leucocyte infiltration in surface and stromal glandular epithelium were observed in both endometrium and endocervix. It was concluded that lipid peroxidation and uterine damage occurs after M. spicata administration in rat uterus. Despite the beneficial effects of M. spicata Labiatae in indigestion, we should also be aware of the toxic effects when it is not used in the recommended fashion, at the recommended dose. PMID- 17120534 TI - Male reproductive impacts of styrene in rat. AB - To determine the effect of styrene on the male reproductive function of rats, male Wistar rats received a daily intraperitoneal (ip) injection of the xenobiotic at a dose of 600 mg/kg body weight. Serum testosterone (T) level was measured in duplicate by radioimmunoassay (RIA). Blood luteinizing hormone (LH) and follicle stimulating hormone (FSH) concentrations were determined using enzyme-linked immunosorbent assay (ELISA). After 10 days of treatment, an increase of the relative weight of the testis occurred, but that of the seminal vesicles and prostate remained unchanged compared to controls injected with an equivalent volume of the vehicle (corn oil). Serum T concentration dropped, while serum hypophyse hormone levels increased. Testicular histological observations revealed a pronounced morphological alteration, with enlarged intracellular spaces, loosening of tissue, and dramatic loss of gametes in the lumen of the seminiferous tubules. Spermatogenesis damage was also confirmed by the decrease in motility and the number of epididymal spermatozoa of treated rats. According to these results, with regard to the lack of a dose response relationship in this study, we may conclude that the testis, precisely the germinal and Sertoli cells, are the major targets for styrene toxicity. PMID- 17120535 TI - Lead and cadmium exposure in children living around a coal-mining area in Yatagan, Turkey. AB - The study was designed to determine asymptomatic lead poisoning prevalence and cadmium exposure of preschool children living in a coal-mining area in Yatagan, Mugla, Turkey. The research was conducted between May and June 2002. The study included 236 children (53.4% female and 46.6% male) who were identified among the healthy children between the ages of 6 months and 6 years, using a systematic sampling method, from the records of the local medical centre of Yatagan. Assessments of the levels of blood lead and cadmium were performed by an atomic absorption spectrophotometer, in the Department of Biophysics, Cerrahpasa Medical Faculty. The mean age of females and males were 49+/-18 and 43+/-19 months, respectively. The mean blood lead level was 33.8+/-15.6 microg/dL in females and 38.8+/-16.0 microg/dL in males. The mean blood lead level of the males was significantly higher than the females (P<0.05). Correlation analysis showed a statistically significant negative correlation between blood lead level and age in both sex groups (r= -0.367, P<0.001). The blood lead level was found to be > 10 microg/dL in 95.7% and >20 microg/dL in 87.6% of all children. The mean blood cadmium level of all children was 1.31+/-0.72 microg/dL. The blood cadmium level was found to be >0.5 microg/dL, which is considered to be toxic, in 85% of all children. The difference in blood cadmium levels between sexes was not significant. A statistically significant negative correlation was found between blood cadmium level and age of all children (r= -0.382, P<0.001). Although it is not possible to understand from this study what proportion of the biological lead and cadmium burden results from mining waste and what proportion comes from other sources, such as paint and gasoline residue deposited in soil and air, these results do indicate that asymptomatic lead poisoning and cadmium exposure are significant problems in children living in the Yatagan area. In conclusion, environmental lead measurements (house dust, soil, drinking water and air) must be performed, the results must be compared with the normal limits, and precautions must be taken if necessary in the Yatagan area. Future public health research efforts should focus on reducing the excessive levels of lead and cadmium in the environment. PMID- 17120536 TI - Exxon Valdez oil no longer a threat? PMID- 17120537 TI - Pesticides lurk in daycare centers. PMID- 17120538 TI - Following mercury in a pristine park. PMID- 17120539 TI - Lichens: ready environmental sensors. PMID- 17120540 TI - Surprising release of metals from CO2 storage. PMID- 17120541 TI - Soybeans resurface in wood glue. PMID- 17120542 TI - Wet electroscrubbers for state of the art gas cleaning. AB - Current trends observed in air pollution control technology are closely related to the development of new, more efficient hybrid systems, i.e., those, which simultaneously utilize two or more physical mechanisms for dust or gaseous contaminants removal. These systems can operate more economically than conventional devices, especially in the removal of PM2.5 particles. The electrostatic scrubber (electroscrubber), discussed in this paper, is one of such types of devices, which combines advantages of electrostatic precipitators and inertial wet scrubbers, and removes many shortcomings inherent to both of these systems operating independently. The electroscrubber is a device in which Coulomb attraction or repulsion forces between electrically charged scrubbing droplets (collector) and dust particles are utilized for the removal of particles from a gas. Unlike wet electrostatic precipitators in which particles are precipitated only on the collection electrode, in electroscrubbers, the collection of dust particles takes place in the entire precipitator chamber. Compared to inertial scrubbers, the electroscrubbers can operate at lower droplet velocities, but the collection efficiency for a single droplet can be larger than 1. The paper reviews the state-of-the-art of wet electrostatic scrubbing (electroscrubbing) technique used for gas cleaning from dust or smoke particles. Three groups of problems are discussed: (1) The fundamental problems concerning the charged dust particle deposition on a charged collector, usually a drop, with a focus on different models describing the process. (2) The experimental works of fundamental importance to our knowledge referring to the scrubbing process, which can be used for validating the theory. (3) The laboratory demonstrations and industrial tests of different constructions of electroscrubbers designed for effective gas cleaning. The electroscrubber is not designed to replace wet or dry electrostatic precipitators but can be used as a complementary device following the last stage of conventional electrostatic precipitator, which helps to remove submicron particles. It was shown in the paper that a higher collection efficiency of an electroscrubber could be obtained for higher values of Coulomb number (i.e., higher electric charges on the collector and particle), and for a Stokes number lower than5 (i.e., low particle-collector relative velocity). PMID- 17120543 TI - Effects of inland nitrogen transport and attenuation modeling on coastal nitrogen load abatement. AB - Modeling of the spatial distribution of nitrogen transport and attenuation from various inland sources and along different hydrological pathways to coastal waters is needed for relevant decisions on effective allocation of measures for coastal nitrogen load abatement. We identify, classify, and quantify uncertainties associated with main discrepancies between spatial process representations in different catchment-scale nitrogen transport-attenuation models. The results show important model differences, indicating scientific disagreement on the realistic spatial process understanding, representation, and quantification in nitrogen transport-attenuation modeling. By further developing solutions for economic optimization of spatially differentiated nitrogen source abatement in coastal catchments, we find this disagreement to considerably affect the economic efficiency of coastal nitrogen load reduction. It may also lead to stakeholder mistrust and conflict and needs to be recognized and handled in environmental policy. PMID- 17120544 TI - The information content of high-frequency environmental monitoring data signals pollution events in the coastal ocean. AB - There are an increasing number of coastal ocean observing systems that deploy new technology for environmental sensing and stream these data in near-real-time to end-users (e.g., scientists and coastal managers) via the worldwide web. The temporal resolution, spatial coverage, and accessibility of these data open up new opportunities for better understanding and managing the coastal ocean, but they also present enormous challenges relative to data processing and data interpretation, particularly in cases where these data are to inform rapid management decision making. Here we demonstrate that changes in surf zone water quality at a popular beach in southern California are signaled by changes in the Fisher Information and Shannon Entropy of high frequency (1/4 min(-1)) measurements of salinity and temperature in the surf zone. These results support the hypothesis that the information content of environmental signals, such as salinity and temperature, can be used to identify changes in the water quality of the coastal ocean. More generally, the approach described here-of using information theory indices calculated from monitoring data as real-time indicators of environmental change-is quite general, and may therefore be applicable to other situations where rapid management decisions are based on high frequency measurements of environmental parameters. PMID- 17120545 TI - Trends in polycyclic aromatic hydrocarbon concentrations in the great lakes atmosphere. AB - Atmospheric polycyclic aromatic hydrocarbon (PAHs) concentrations were measured in both the vapor and particle phases at seven sites near the Great Lakes as a part of the Integrated Atmospheric Deposition Network. Lower molecular weight PAHs, including fluorene, phenanthrene, fluoranthrene, and pyrene, were dominant in the vapor phase, and higher molecular weight PAHs, including chrysene, benzo[a]pyrene, and coronene, were dominant in the particle phase. The highest PAH concentrations in both the vapor and particle phases were observed in Chicago followed by the semiurban site at Sturgeon Point, NY. The spatial difference of PAH concentrations can be explained by the local population density. Long-term decreasing trends of most PAH concentrations were observed in both the vapor and particle phases at Chicago, with half-lives ranging from 3-10 years in the vapor phase and 5-15 years in the particle phase. At Eagle Harbor, Sleeping Bear Dunes, and Sturgeon Point, total PAH concentrations in the vapor phase showed significant, but slow, long-term decreasing trends. At the Sturgeon Point site, which was impacted by a nearby city, particle-phase PAH concentrations also declined. However, most particle-phase PAH concentrations did not show significant long-term decreasing trends at the remote sites. Seasonal trends were also observed for particle-phase PAH concentrations, which were higher in the winter and lower in the summer. PMID- 17120546 TI - PCDD and PCDF emissions from simulated sugarcane field burning. AB - The emissions from simulated sugarcane (Saccharum officinarum) field burns were sampled and analyzed for polychlorinated dibenzodioxins and dibenzofurans (PCDDs and PCDFs). Sugarcane leaves from Hawaii and Florida were burned in a manner simulating the natural physical dimensions and biomass density found during the practice of preharvest field burning. Eight composite burn tests consisting of 3 33 kg of biomass were conducted, some with replicate samplers. Emission factor calculations using sampled concentration and measured mass loss compared well to rigorous carbon balance methods commonly used in field sampling. The two sources of sugarcane had distinctive emission levels, as did tests on separate seasonal gatherings of the Florida sugarcane. The average emission factor for two tests of Hawaii sugarcane was 253 ng toxic equivalents (TEQ)/kg of carbon burned (ng TEQ/kg(Cb)) (rsd = 16%) and for two gatherings of Florida sugarcane was 25 ng TEQ/kg(Cb) (N = 4, rsd = 50%) and 5 ng TEQ/kg(Cb) (N = 2, rsd = 91%). The Hawaii sugarcane, as well as most of the Florida sugarcane, had emission values which were well above the value of 5 ng TEQ/kg(Cb) commonly attributed to biomass combustion. Application of this emission factor range to the amount of U.S. sugarcane fields burned suggests that this practice may be a relatively minor source of PCDDs and PCDFs in the U.S. national inventory, but the limited sample size and range of results make this conclusion tenuous. PMID- 17120547 TI - TEM study of PM2.5 emitted from coal and tire combustion in a thermal power station. AB - The research presented here was conducted within the scope of an experiment investigating technical feasibility and environmental impacts of tire combustion in a coal-fired power station. Previous work has shown that combustion of a coal+tire blend rather than pure coal increased bulk emissions of various elements (e.g., Zn, As, Sb, Pb). The aim of this study is to characterize the chemical and structural properties of emitted single particles with dimensions <2.5 microm (PM2.5). This transmission electron microscope (TEM)-based study revealed that, in addition to phases typical of coal fly ash (e.g., aluminum silicate glass, mullite), the emitted PM2.5 contains amorphous selenium particles and three types of crystalline metal sulfates never reported before from stack emissions. Anglesite, PbSO4, is ubiquitous in the PM2.5 derived from both fuels and contains nearly all Pb present in the PM. Gunningite, ZnSO4-H2O, is the main host for Zn and only occurs in the PM derived from the coal+tire blend, whereas yavapaiite, KFe3+(SO4)2, is present only when pure coal was combusted. We conclude that these metal sulfates precipitated from the flue gas, may be globally abundant aerosols, and have, through hydration or dissolution, a major environmental and health impact. PMID- 17120548 TI - Fate, partitioning, and mass loading of polybrominated diphenyl ethers (PBDEs) during the treatment processing of municipal sewage. AB - Sewage treatment plant (STP) effluents are likely a major source of contamination for PBDEs, especially in the receiving water bodies of local aquatic environments surrounding the location of these discharges. Congeners of the pentaBDE mixture, 2,2,',4,4'-tetrabromodiphenyl ether (BDE47), 2,2,',4,4',5-pentabromodiphenyl ether (BDE99), 2,2,',4,4',6-pentabromodiphenyl ether (BDE100), 2,2,',4,4',5,5' hexabromodiphenyl ether (BDE153), and 2,2,',4,4',5,6'-hexabromodiphenyl ether (BDE154), are of great environmental concern in North America due to their persistence, potential for bioaccumulation, and >97% use of the global production of the mixture in the region. Detailed characterization of the distribution of eight PBDE congeners (2,4,4'-tribromoDE (BDE28) and BDE47, 99, 100, 138, 153, 154, and 183) was carried out at five sites along the treatment process at an activated sludge-type secondary treatment municipal STP facility. PentaBDE mixture congeners, sigma5PBDE (sum of BDE47, 99, 100, 153, and 154) accounted for >98% of the total (sigma 8) PBDE concentration at all sites, with over 80% of the composition being BDE47 and BDE99. Presence of dissolved organic matter affected the mobility of PBDEs during the initial stages of the treatment process. About 9% of the influent mass of sigma5PBDE to the facility is estimated to be discharged into the Little River (leading to the Detroit River) with the final effluent, resulting in an estimated mass loading of approximately 0.7 kg/year. The total mass loading of sigma5PBDE to the Detroit River is expected to be much larger as effluent from this facility accounts for <10% of the total STP discharges to the river. PMID- 17120549 TI - Detailed polybrominated diphenyl ether (PBDE) congener composition of the widely used penta-, octa-, and deca-PBDE technical flame-retardant mixtures. AB - Polybrominated diphenyl ethers (PBDEs) have been widely used to flame-retard products common in homes and the workplace, and subsequently, they have become widely dispersed in the environment. Detailed compositional knowledge of these complex PBDE mixtures is crucial to a fuller understanding of their toxicological potencies and environmental fate due to selective congener biomagnification, degradation, and transport. Utilizing recenttechnical enhancements and newly available commercial standards, we developed a method capable of analyzing a larger suite of mono- through deca-BDEs. We then characterized the congener composition of six common technical flame-retardant mixtures: two penta-BDE products (DE-71 and Bromkal 70-5DE) two octa-BDE products (DE-79 and Bromkal 79 8DE) and two deca-BDE products (Saytex 102E and Bromkal 82-0DE). PBDEs were analyzed by gas chromatography/mass spectrometry (GC/MS). Structural conformations based on fragmentation patterns and molecular ions were established by electron-capture negative ionization (ECNI) and electron ionization (El). Sixty-four commercially available PBDE standards were chromato-graphed on two GC columns (DB-1HT and DB-5HT) and relative retention indexes (RRI) calculated. Thirty-nine PBDEs were identified in these products, 29 at concentrations >0.02% by weight. Of these, 12 previously unreported congeners have been confirmed as commercial mixture components. Four of these congeners were detected >0.02% w/w (BDE-144, -171, -180, and -201) and three (BDE-75, -184, and -194) at <0.02%. Five other congeners (four <0.02% by weight) were tentatively identified based on their molecular ion and ECNI fragmentation in the absence of corresponding analytical standards. PMID- 17120550 TI - Low molecular weight dicarboxylic acids, ketoacids, and dicarbonyls in the fine particles from a roadway tunnel: possible secondary production from the precursors. AB - Low molecular weight dicarboxylic acids (DCAs), ketoacids, and alpha-dicarbonyls have been determined for the PM2.5 samples in a Hong Kong roadway tunnel, using a water extraction followed by a butyl ester and/or dibutyl acetal derivatization technique. For the most wintertime sampling runs, outlet and inlet concentrations of the quantified compounds were found to be quite similar (ca. 10% differences), leading to the conclusion that direct emissions of the organic compounds are insignificant from vehicles in hot-stabilized operations although vehicular emissions can provide important precursors to them. In contrast, a significant concentration increase of most compounds was observed at the outlet station compared to the inlet station in the summertime runs, which might be explained by the secondary production of aerosols in the tunnel. The organic compounds studied comprised a small fraction (<1%) of aerosol organic carbon (OC). In winter, their abundances relative to that of OC in outlet samples were found to be significantly less than those in inlet samples. On the basis of the summer data, apparent secondary production factors of the compounds were calculated, which indicate that adipic and m-phthalic acids can be favorably formed in the tunnel. However, like other DCAs, direct emissions of adipic, m-phthalic, and p-phthalic acids from automobiles are suggested to be insignificant. PMID- 17120551 TI - Mercury in soils, lakes, and fish in Voyageurs National Park (Minnesota): importance of atmospheric deposition and ecosystem factors. AB - Concentrations of methylmercury in game fish from many interior lakes in Voyageurs National Park (MN, U.S.A.) substantially exceed criteria for the protection of human health. We assessed the importance of atmospheric and geologic sources of mercuryto interior lakes and watersheds within the Park and identified ecosystem factors associated with variation in methylmercury contamination of lacustrine food webs. Geologic sources of mercury were small, based on analyses of underlying bedrock and C-horizon soils, and nearly all mercury in the 0- and A-horizon soils was derived from atmospheric deposition. Analyses of dated sediment cores from five lakes showed that most (63% +/- 13%) of the mercury accumulated in lake sediments during the 1900s was from anthropogenic sources. Contamination of food webs was assessed by analysis of whole, 1-year-old yellow perch (Perca flavescens), a regionally important prey fish. The concentrations of total mercury in yellow perch and of methylmercury in lake water varied substantially among lakes, reflecting the influence of ecosystem processes and variables that affect the microbial production and abundance of methylmercury. Models developed with the information-theoretic approach (Akaike Information Criteria) identified lake water pH, dissolved sulfate, and total organic carbon (an indicator of wetland influence) as factors influencing methylmercury concentrations in lake water and fish. We conclude that nearly all of the mercury in fish in this seemingly pristine PMID- 17120552 TI - Pesticide measurements from the first national environmental health survey of child care centers using a multi-residue GC/MS analysis method. AB - The U.S. Department of Housing and Urban Development, in collaboration with the U.S. Consumer Product Safety Commission and the U.S. Environmental Protection Agency, characterized the environments of young children (<6 years) by measuring lead, allergens, and pesticides in a randomly selected nationally representative sample of licensed institutional child care centers. Multi-stage sampling with clustering was used to select 168 child care centers in 30 primary sampling units in the United States. Centers were recruited into the study by telephone interviewers. Samples for pesticides, lead, and allergens were collected at multiple locations in each center by field technicians. Field sampling was conducted from July through October 2001. Wipe samples from indoor surfaces (floors, tabletops, desks) and soil samples were collected at the centers and analyzed using a multi-residue GC/MS analysis method. Based on the questionnaire responses, pyrethroids were the most commonly used pesticides among centers applying pesticides. Among the 63% of centers reporting pesticide applications, the number of pesticides used in each center ranged from 1 to 10 and the frequency of use ranged from 1 to 107 times annually. Numerous organophosphate and pyrethroid pesticides were detected in the indoor floor wipe samples. Chlorpyrifos (0.004-28 ng/cm2), diazinon (0.002-18 ng/cm2), cis-permethrin (0.004 3 ng/cm2), and PMID- 17120553 TI - Brominated flame retardants and halogenated phenolic compounds in North American west coast bald eaglet (Haliaeetus leucocephalus) plasma. AB - We report on the identity, characterization, and spatial trends of several brominated flame retardants and hydroxylated (OH-) and methoxylated (MeO-) organohalogen contaminants in bald eagle (Haliaeetus leucocephalus) nestling plasma collected from sites along the west coast of North America. Samples were from four southwestern British Columbia (BC) locations, a reference site in northern BC (Fort St. James; FSJ), and from Santa Catalina Island, CA (SCI), an area of high DDT and PCB contamination. Mean concentrations of sigma polybrominated diphenyl ether (sigma PBDE (8 congeners monitored); 1.78-8.49 ng/g), sigma OH-polychlorinated biphenyl (sigma OH-PCB (30 congeners monitored); 0.44-0.87 ng/g), and sigma OH-PBDE (14 congeners monitored; 0.31-0.92 ng/g) were similar in eaglets from southwestern BC yet lower than for SCl and significantly higher than for FSJ. Dominant PBDE congeners were BDE47, BDE99, and BDE100, but SCl eaglets also contained low levels of higher brominated congeners. 4-OH-CB187 and 4'-OH-CB202 accounted for 65-100% of sigma OH-PCB in all BC eaglets, with 4' OH-CB202 as well as 3'-OH-CB138 and 4-OH-CB146 dominating in SCl eaglets. Ostensibly of biogenic origin, 6'-OH-BDE49 and 6-OH-BDE47 were found in BC nestlings. Only 4'-OH-BDE49 (2.10 ng/g) was found in SCl eaglets. MeO-PBDEs and total hexabromocyclododecane (HBCD) were not found in any birds, but the polybrominated biphenyl BB101 was detected in southwestern BC samples. This study demonstrates that west coast North American bald eagles contain previously unreported organohalogens, which have the potential to impact the health and survival of these raptors. PMID- 17120554 TI - Hydroxy-PCBs, PBDEs, and HBCDDs in serum from an elderly population of Swedish fishermen's wives and associations with bone density. AB - Lack of human exposure data is frequently reported as a critical gap in risk assessments of environmental pollutants, especially regarding "new" pollutants. The objectives of this study were to assess serum levels of the persistent 2,2',4,4',5,5'-hexachlorobiphenyl (CB-153), hydroxylated polychlorinated biphenyl metabolites (OH-PCBs), polybrominated diphenyl ethers (PBDEs), and hexabromocyclodode-canes (HBCDDs) in a group of Swedish middle-aged and elderly women expected to be relatively highly exposed, and to evaluate the impact of potential determinants (e.g., fish intake, age) for the inter-individual variation, as well as to investigate the association between these pollutants and bone density. No associations were found between bone mineral density or biochemical markers of bone metabolism and the analyzed environmental pollutants. Relatively high levels of CB-153 (median 260 ng/g fat) and sigma 3-OH-PCBs (median 1.7 ng/mL serum), and low concentrations of sigma6PBDEs (median 3.6 ng/g fat) were determined. Total level of HBCDDs in serum was quantified by gas chromatography with mass spectrometric detection (median 0.5 ng/g fat). HBCDD diastereomeric and enantiomeric patterns were determined by liquid chromatography with mass spectrometric detection. The dominating stereoisomer was (-)alpha HBCDD, but 1-3% of gamma-HBCDD was also detected in the serum samples. PMID- 17120555 TI - Kinetics of microbial reduction of Solid phase U(VI). AB - Sodium boltwoodite (NaUO2SiO3OH x 1.5 H2O) was used to assess the kinetics of microbial reduction of solid-phase U(VI) by a dissimilatory metal-reducing bacterium (DMRB), Shewanella oneidensis strain MR-1. The bioreduction kinetics was studied with Na-boltwoodite in suspension or within alginate beads in a nongrowth medium with lactate as electron donor at pH 6.8 buffered with PIPES. Concentrations of U(VI)tot and cell number were varied to evaluate the coupling of U(VI) dissolution, diffusion, and microbial activity. Microscopic and spectroscopic analyses with transmission electron microscopy (TEM), energy dispersive spectroscopy (EDS), and laser-induced fluorescence spectroscopy (LIFS) collectively indicated that solid-phase U(VI) was first dissolved and diffused out of grain interiors before it was reduced on bacterial surfaces and/or within the periplasm. The kinetics of solid-phase U(VI) bioreduction was well described by a coupled model of bicarbonate-promoted dissolution of Na-boltwoodite, intragrain uranyl diffusion, and Monod type bioreduction kinetics with respect to dissolved U(VI) concentration. The results demonstrated that microbial reduction of solid-phase U(VI) is controlled by coupled biological, chemical, and physical processes. PMID- 17120556 TI - Impact of redox conditions on arsenic mobilization from tailings in a wetland with neutral drainage. AB - More than 80 years of silver mining in the Cobalt area (Ontario, Canada) has led to widespread contamination of water with arsenic. The objective of this study was to determine the impact of changes in redox conditions on the stability of As in samples collected from a tailings wetland in the historic mining camp. Dissolved metal concentrations were monitored while tailings samples (approximately 1300 mg of As kg(-1), pH 7.4) were subjected to 30 days of reduction. Reoxidation of the samples was accomplished by air drying. The As oxidation states in the original, reduced, and reoxidized samples were determined using X-ray absorption spectroscopy (XAS). Arsenic speciation was affected by changes in redox conditions, resulting in rapid mobilization of As during reduction. Glucose input had a significant impact on the dissolution and speciation of As, suggesting that the As transformation was microbially mediated. When carbon was not limiting, the combination of reducing conditions and lower pH favored the formation of As(-I) species. PMID- 17120557 TI - Mechanisms of gold bioaccumulation by filamentous cyanobacteria from gold(III) chloride complex. AB - The mechanisms of gold bioaccumulation by cyanobacteria (Plectonema boryanum UTEX 485) from gold(III)-chloride solutions have been studied at three gold concentrations (0.8,1.7, and 7.6 mM) at 25 degrees C, using both fixed-time laboratory and real-time synchrotron radiation absorption spectroscopy (XAS) experiments. Interaction of cyanobacteria with aqueous gold(III)-chloride initially promoted the precipitation of nanoparticles of amorphous gold(I) sulfide at the cell walls, and finally deposited metallic gold in the form of octahedral (111) platelets (approximately 10 nm to 6 microm) near cell surfaces and in solutions. The XAS results confirm that the reduction mechanism of gold(III)-chloride to metallic gold by cyanobacteria involves the formation of an intermediate Au(I) species, gold(I)-sulfide. PMID- 17120558 TI - Long-term aging of copper added to soils. AB - Aging refers to the processes by which the mobility and bioavailability/toxicity of metals added to soil decline with time. Although long-term aging is a key process that needs to be considered in the risk assessment of metals in field soils, the long-term aging of Cu added to soils has not been studied in detail. In this study, the effects of aging on Cu isotopic exchangeability, total Cu in soil pore water, pore water free Cu2+ activity, and "available" Cu measured by the technique of diffusive gradient in thin film (DGT-Cu) were investigated in 19 European soils at two total Cu concentrations shown to inhibit plant (tomato) growth by 10 and 90%. After addition of Cu, the soils were leached, incubated outdoors, and sampled regularly over a 2-year period. The results showed that when water soluble Cu was added to soils, concentrations of Cu determined by each of the techniques tended to decrease rapidly initially, followed by further decreases at slow rates. Soil pH was a vital factor affecting the aging rate of Cu added to soils. The relatively low solubility products and low isotopic exchangeabilities of Cu in calcareous soils immediately after addition of soluble Cu2+ suggested Cu2+ probably precipitated in these soils as Cu2(OH)2CO3 (malachite) and Cu(OH)2. Isotopic dilution was found to be a robust technique for measuring rates of long-term aging reactions. A semi-mechanistic model was developed to describe the rate and extent of Cu aging across soils as affected by soil pH and other physicochemical parameters. Although not measured directly, it is inferred from soil physicochemical controls on Cu aging that processes of precipitation/nucleation of Cu in soils and hydrolysis of Cu2+ followed by a diffusion process controlled the decrease in Cu availability with time. The model was validated by testing it against field soils with different contamination histories and was found to successfully predict the isotopic exchangeability of Cu added to soils based on two parameters: soil pH and time. PMID- 17120559 TI - Oligomer formation in evaporating aqueous glyoxal and methyl glyoxal solutions. AB - Glyoxal and methyl glyoxal are common secondary atmospheric pollutants, formed from aromatic and terpene precursors. Both compounds are extremely water-soluble due to dihydrate formation and partition into cloudwater. In this work, FTIR-ATR and mass measurements indicate that both compounds remain primarily in the condensed phase due to oligomer formation when aqueous solution droplets are evaporated, regardless of concentration (> or = 1 mM) or, for glyoxal, droplet evaporation rate. FTIR spectral analyses suggestthat oligomer formation is triggered by conversion from dihydrate to monohydrate forms, which are still nonvolatile but contain reactive carbonyl groups. Methyl glyoxal hemiacetal formation is observed by changes in the C-0/C=0 stretch peak area ratio. The formation of glyoxal oligomers is detected by a dramatic shift of the C-0 stretching peak toward low frequencies. Glyoxal oligomer peaks at 1070 cm(-1), 950 cm(-1), and 980 cm(-1) are assigned to free C-OH stretch, dioxolane-linked C OC asymmetric stretch, and tentativelyto non-dioxolane-linked C-OC stretches, respectively. Acids have little effect on glyoxal oligomer formation; however, base interrupts oligomer formation by catalyzing glyoxal hydration and disproportionation to glycolic acid. Since glyoxal and methyl glyoxal are commonly found in cloudwater and are expected to remain largely in the aerosol phase when cloud droplets evaporate, this process may be a source of secondary organic aerosol by cloud processing. PMID- 17120560 TI - Prion degradation in soil: possible role of microbial enzymes stimulated by the decomposition of buried carcasses. AB - This study is part of a European project focused on understanding the biotic and abiotic mechanisms involved in the retention and dissemination of transmissible spongiform encephalopathies (TSE) infectivity in soil in order to propose practical recommendations to limit environmental contamination. A 1-year field experiment was conducted with lamb carcasses buried in a pasture soil at three depths (25, 45, and 105 cm). Microbial community response to carcasses was monitored through the potential proteolytic activity and substrate induced respiration (SIR). Soil above carcasses and control soil exhibited low proteolytic capacity, whatever the depth of burial. Contrastingly, in soil beneath the carcasses, proteolysis was stimulated. Decomposing carcasses also stimulated SIR, i.e., microbial biomass, suggesting that proteolytic populations specifically developed on lixiviates from animal tissues. Decomposition of soft tissues occurred within 2 months at subsurface while it lasted at least 1 year at deeper depth where proteolytic activities were season-dependent. The ability of soil proteases to degrade the beta form of prion protein was shown in vitro and conditions of burial relevant to minimize the risk of prion protein dissemination are discussed. PMID- 17120561 TI - Hydrolytic stability of terephthaloyl chloride and isophthaloyl chloride. AB - The phthaloyl chloride isomers, terephthaloyl chloride (TCl) and isophthaloyl chloride (ICl), are high production volume chemicals used in polymers to impartflame resistance, chemical resistance, and temperature stability and as water scavengers. In these studies, we determined the hydrolytic stability of TCl and ICl and their hydrolysis products in aqueous solutions. Hydrolysis rates for TCl and ICl were initially determined by gas chromatography/flame ionization detection in water buffered at pH 4.0, 7.0, and 9.0 and 0 degrees C for up to 30 min. Subsequent studies determined the products from TCl and ICl hydrolysis. The parent phthaloyl chlorides (TCl and ICl), their intermediate hydrolysis products (designated as the "half-acids"), and their stable hydrolysis products (terephthalic acid (TPA) and isophthalic acid (IPA)) were determined by high performance liquid chromatography. The half-lives (t(1/2)) of TCl and ICl ranged from an average of 1.2 to 2.2 min and from 2.2 to 4.9 min, respectively, at pH 4 9 and 0 degrees C. The observed first-order rate constants (k(obs)) ranged from an average of 530 to 1100 (x 10(5) s(-1)) for TCl and 240 to 520 (x 10(5) s(-1)) for ICl. Both phthaloyl chlorides formed their respective short-lived intermediates, in which one of the two carboxylic acid chloride functionalities reacts with water to form the carboxylic acid ("half-acid"). Subsequently, the half-acids underwent further hydrolysis so that greater than 90% of the initial phthaloyl chloride hydrolyzed in less than 60 min at 0 degrees C. The hydrolysis products TPA and IPA were hydrolytically stable, undergoing no further transformations after 20 min at pH 7 and 25 degrees C. This work demonstrated that TCl, ICl, and their respective half-acids will not be persistent in aqueous systems for a time sufficient to have a sustained toxicological effect on aquatic organisms (less than 1 h). Performing additional aquatic toxicity studies, biodegradation studies, and potentially mammalian studies on TCl and ICl are unnecessary since the existing information on TPA and IPA with the hydrolysis data presented here is sufficient to address questions on the fate and effects of these two substances in aqueous environments. PMID- 17120562 TI - Transport of rodlike colloids through packed beds. AB - The effect of colloid shape on filtration rates in porous media was examined by constructing particles with different aspect ratios and measuring their retention in packed beds. Spherical polystyrene latex microspheres (1.0-microm diameter) were heated, stretched to the desired aspect ratio (2:1 and 3:1, with a 1:1 control), and quickly cooled. These particles were injected into minicolumns containing glass beads (40-microm diameter) in solutions at two different ionic strengths (IS = 1 and 100 mM). The measured retentions increased with aspect ratio in both IS solutions. The zeta-potentialsfor all three aspect ratios were indistinguishable, and no charge nonuniformity was measured for any of the samples. Thus, the data supportthat changes in retention resulted from the different aspect ratios rather than from different surface chemistries. Interpretation of the retention data in terms of a collision efficiency (alpha) showed an increase with aspect ratio in both IS solutions, and for 1 mM the alpha increased from 0.011 (1:1) to 0.095 (2:1) to 0.26 (3:1). These results demonstrate for the first time the direct impact of particle shape on retention in porous media. Our findings have important implications for the transport of particles with high aspect ratios, such as rod-shaped bacteria, and for the modeling of such transport. PMID- 17120563 TI - The impact of particle-bound cadmium on bioavailability and bioaccumulation: A pragmatic approach. AB - Studying the bioavailability of sediment-bound contaminants is complicated by many reasons, such as the variable composition of the particles, their temporal variations, the low levels of contaminant concentrations, their partitioning between diverse aqueous and particulate phases, and the variety of uptake routes that may involved with the biota. Therefore, simple and innovative methodologies should be tested as analogues for natural sediments. Among them, a diverse selection of artificial particles with well-defined surface properties, in the presence and absence of commercially available humic acids, has been proposed and used to investigate the bioavailability of several organic pollutants. For this work, this model was applied to investigate the uptake and accumulation of cadmium by the freshwater oligochaete Lumbriculus variegatus. The results showed that the uptake of the metal depended on the free dissolved Cd(II) species, while the contribution from the particles was negligible. Thus, the extent of cadmium bioaccumulated from each test system could be predicted as a function of the rate of absorption of the free dissolved Cd(II) species. These species were calculated either from the particle-water partition coefficients, or by using the MINEQL+ computer program. In general, the estimated accumulation levels were in good agreement with the experimental results. PMID- 17120564 TI - Comparison of chemical approaches for assessing bioavailability of sediment associated contaminants. AB - Two chemical approaches, Tenax extraction and matrix solid-phase microextraction (matrix-SPME), were compared to assess the bioavailability of hydrophobic contaminants from sediment. Hexachlorobiphenyl, DDE, permethrin, chlorpyrifos, and phenanthrene were individually spiked into two sediments differing in physical characteristics. Bioaccumulation was determined by exposing the oligochaete, Lumbriculus variegatus, to the spiked sediments. The rapidly desorbing fraction from Tenax extraction at 6 h and fiber concentration at 14 d from the matrix-SPME were compared for predicting bioaccumulation. Further, a comparison between laboratory-spiked and field-contaminated sediments was conducted. A regression between the rapidly desorbed sediment concentration at 6 h and the amount bioaccumulated across compounds and sediments described 94% of the variation in the data when phenanthrene was excluded. Phenanthrene was excluded because of complications due to a combination of biotransformation and rapid elimination during the sampling process. Contaminant accumulation by L. variegatus also correlated well with matrix-SPME fiber concentrations, accounting for 92% of the variation in the data, again excluding phenanthrene. Both chemical methods provided matrix- and chemical-independent estimations of bio-accumulation for hydrophobic contaminants without extensive biotransformation. PMID- 17120565 TI - Application of heuristic optimization techniques and algorithm tuning to multilayered sorptive barrier design. AB - Although heuristic optimization techniques are increasingly applied in environmental engineering applications, algorithm selection and configuration are often approached in an ad hoc fashion. In this study, the design of a multilayer sorptive barrier system served as a benchmark problem for evaluating several algorithm-tuning procedures, as applied to three global optimization techniques (genetic algorithms, simulated annealing, and particle swarm optimization). Each design problem was configured as a combinatorial optimization in which sorptive materials were selected for inclusion in a landfill liner to minimize the transport of three common organic contaminants. Relative to multilayer sorptive barrier design, study results indicate (i) the binary-coded genetic algorithm is highly efficient and requires minimal tuning, (ii) constraint violations must be carefully integrated to avoid poor algorithm convergence, and (iii) search algorithm performance is strongly influenced by the physical-chemical properties of the organic contaminants of concern. More generally, the results suggest that formal algorithm tuning, which has not been widely applied to environmental engineering optimization, can significantly improve algorithm performance and provide insight into the physical processes that control environmental systems. PMID- 17120566 TI - Mechanism and elimination of a water vapor interference in the measurement of ozone by UV absorbance. AB - A water vapor interference in ozone measurements by UV absorption was investigated using four different ozone monitors (TEI models 49 and 49C, Dasibi model 1003-AH, and a 2B Technologies model 202 prototype). In the extreme case of step changes between 0 and 90% relative humidity (RH), a large interference in the range of tens to hundreds of ppbv was found for all instruments tested, with the magnitude and sign depending on the manufacturer and model. Considering that water vapor does not absorb at the wavelength of the Hg lamp (253.7 nm) used in these instruments, another explanation is required. Based on experimental evidence and theoretical considerations, we conclude that the water vapor interference is caused by humidity effects on the transmission of uncollimated UV light through the detection cell. The ozone scrubber acts as a water reservoir, either adding or removing water from the air sample, thereby modulating the detector signal and producing a positive or negative offset. It was found for the 2B Technologies ozone monitor that use of a 1-m length of Nafion tubing just prior to the entrance to the detection cell reduces the water vapor interference to negligible levels (+/- 2 ppbv for step changes between 0 and 90% RH) while quantitatively passing ozone. PMID- 17120567 TI - An evaluation of DGT performance using a dynamic numerical model. AB - A numerical model of the transport and dynamics of metal complexes in the resin and gel layers of a DGT (diffusive gradients in thin films) device was developed and used to investigate how the chelating resin and metal-ligand complexes in solution affect metal uptake. Decreasing the stability constant or concentration of the binding resin increases the competition for free metal ions by ligands in solution, lowering the rate of mass uptake. Such effects would be rarely observed for moderately or strongly binding resins (K> 10(12)), including Chelex, which out-compete labile ligands in solution. With weakly binding resins, strongly bound solution complexes can diffuse into the resin layer before a measurable amount of dissociation occurs, such that concentrations of bound metal at the rear and front surfaces of the resin layer are equal. With more strongly binding resins, metal mainly binds to the front surface of the resin. Only complexes with the largest binding constants penetrate the gel layer containing Chelex, buttheir lack of lability means thatthe DGT sensitivity to the complex is, in any case, very low. The slow diffusion of complexes, such as those of fulvic acids, which increases the time required to establish steady state, compromises the use of the simple DGT equation. Errors are negligible for 24 h deployments, when diffusive layer thicknesses are less than 1 mm, but 3 day deployments are required to ensure accuracy with 2.4 mm thick layers. The extent to which the commonly used equation, that accounts for the concentration and diffusion of metal-complex species, overestimates DGT uptake if the rate of dissociation is slow, was estimated. PMID- 17120568 TI - Intercomparison of thermal and optical measurement methods for elemental carbon and black carbon at an urban location. AB - Despite intensive efforts during the past 20 years, no generally accepted standard method exists to measure black carbon (BC) or elemental carbon (EC). Data on BC and EC concentrations are method specific and can differ widely (e.g. Schmid et al., 2001, ten Brink et al., 2004). In this study, a comprehensive set of methods (both optical and thermal) is compared. Measurements were performed under urban background conditions in Vienna, Austria, a city heavily impacted by diesel emissions. Filter and impactor samples were taken during 3 weeks in summer 2002 and analyzed for EC with thermal methods: a modified Cachier method (Cachier et al., 1989), a thermal-optical method (Schmid et al., 2001), and the VDI method (VDI, 1996); for BC with optical methods: a filter transmission method and the integrating sphere method (Hitzenberger et al., 1996); and for total carbon (TC) with a combustion method (Puxbaum and Rendl, 1983). The online methods aethalometer (Hansen et al., 1984) and the multiangle absorption photometer MAAP (Petzold et al., 2002) to measure BC were also used. The average values of BC and EC obtained with the methods agreed within their standard deviations. A conversion table was set up to allow comparisons between data measured elsewhere under urban background conditions (with similar source characteristics) with different instruments. An approach to estimate the absorption coefficient from attenuation data is derived so that existing records of aethalometer data in urban environments may be used to obtain also the absorption coefficients. PMID- 17120569 TI - Use of lichens as pollution biomonitors in remote areas: comparison of PAhs extracted from lichens and atmospheric particles sampled in and around the Somport tunnel (Pyrenees). AB - Lichens of the species Parmelia sulcata were collected from sites on both sides of the Somport tunnel (which links France and Spain) and atmospheric particles were collected by air samplers installed within and on either side of the tunnel. Polycyclic aromatic hydrocarbons (PAHs) in the lichen and particle samples were then extracted, identified, quantified, and compared to evaluate the potential utility of lichens as pollution biomonitors in remote areas. The origin of the PAHs was also assessed using the Phe/Ant, Flu/Pyr, Ant/Ant+Phe, Flu/Flu+Pyr, and BaA/BaA+Chr concentration ratios. The total concentration of 16 priority PAHs ranged from 6.79 to 23.3 microg/g in particles outside the tunnel, from 18.3 to 265.2 microg/g in particles inside the tunnel, and from 0.91 to 1.92 microg/g in the lichen samples. The PAH ratios found in the lichens and particulate matter indicate that they were of pyrogenic origin and that road traffic was a major contributor. Results from the lichen samples suggest that they may be excellent biomonitors of pollution in remote areas. PMID- 17120570 TI - Magnitude and directional measures of water and Cr(VI) fluxes by passive flux meter. AB - A new configuration of the passive fluxmeter (PFM) is presented that provides for simultaneous measurements of both the magnitude and the direction of ambient groundwater specific discharge qo and Cr(VI) mass flux J(Cr). The PFM is configured as a cylindrical unit with an interior divided into a center section and three outer sectors, each packed with a granular anion exchange resin having high sorption capacity for the Cr(VI) oxyanions CrO4(2-) and HCrO4-. The sorbent in the center section is preloaded with benzoate as the "resident" tracer. Laboratory experiments were conducted in which PFMs were placed in porous packed bed columns, through which was passed a measured volume of synthetic groundwater containing Cr(VI). During the deployment period, some of the resident tracer is depleted while the Cr(VI) is sorbed. The resin was then removed from the four sectors separately and extracted to determine the "captured" mass of Cr(VI) and the residual mass of the resident tracer in each. Cumulative specific discharge, q0t, values were assessed using the residual mass of benzoate retained in the center section. The direction of this discharge theta was ascertained from the mass distribution of benzoate intercepted and retained in the outer three sections of the PFM. Cumulative chromium fluxes, J(Cr)t, were quantified using the total Cr(VI) mass intercepted and retained on the PFM. Experiments produced an average measurement error for direction theta of 3 degrees +/- 14 degrees, while the average measurement errors for q0 and J(Cr) were, respectively, -8% +/- 15% and -12% +/- 23%. Results demonstrate the potential utility of the new PFM configuration for characterizing groundwater and contaminant fluxes. PMID- 17120571 TI - Field comparison of particulate PAH measurements using a low-flow denuder device and conventional sampling systems. AB - Polyaromatic hydrocarbons (PAHs) are complex carbonaceous compounds emitted to the atmosphere by various combustion processes. Because the toxicity of many of them is now well recognized and documented, the determination of their atmospheric concentrations is of great interest to better understand and develop future atmospheric pollution control strategies. Hence, a common sampling protocol has to be defined to homogenize the results. With this goal in mind, field studies were carried out under different environmental conditions (74 samples) by simultaneously operating both a conventional sampler and a sampler equipped with a denuder tube upstream from the filter. The experimental results presented in this work show that the atmospheric particulate PAH concentrations are underestimated at least by a factor of 2 using a conventional sampler. The discrepancy between the two kinds of samplers used varied a lot from one compound to another and from one field campaign to another. This discrepancy may be explained by a simple degradation of particulate PAH in the natural atmosphere and on the filter. This is particularly worrisome because, based on the results presented in this work, the atmospheric PAH concentrations measured using conventional samplers not equipped with an ozone trap can underestimate the PAH concentration by more than 200%. This is especially true when the samples are collected in the vicinity of the point source of particulate PAHs and for highly reactive compounds such as benzo[a]pyrene. PMID- 17120572 TI - Analysis for perfluorocarboxylic acids/anions in surface waters and precipitation using GC--MS and analysis of PFOA from large-volume samples. AB - The presence of perfluorocarboxylates (PFCAs) in the environment is of increasing concern, following the discovery of perfluoroalkyl acids (PFAs) in wildlife and human samples. Here we report a method forthe determination of (C2-C9) PFCAs by preparing the 2,4-difluoroanilides of the acids and analyzing by using GC-MS. Detector response was linear over the range 0.1 -1000 pg of each perfluoroalkyl anilide. A complete suite of PFCAs can be analyzed in an individual sample with the PFCAs detected at levels similar to or lower than those determined by other methods. For a comparison between the present method and the more common LC-MS/MS method, 10 replicates of a sewage treatment plant discharge were analyzed for perfluoro-octanoic acid (PFOA) using both methods. Results were nearly identical with low standard deviation (GC-MS 30.9 +/- 1.88 ng/L; while the LC-MS/MS 34.7 +/ 3.05 ng/L). PFCA concentrations for water samples collected from depth profiles in mid-Lake Ontario were analyzed by GC-MS with most PFCAs (C2-C8) present above the detection limit (0.5 ng/L). Major PFCAs were trifluoroacetate (TFA) (100 ng/L) and perfluorobutanoate (PFBA) (> 5 ng/L). Results for PFOA (2.5 ng/L) were in good agreement with recent analyses by LC-MS/MS. PFCAs were also detected in the precipitation samples at concentrations lower than those of the samples from the lake profiles or sewage treatment plants (STPs) effluent. Since PFOA levels may be less than the lower detection limit (<0.5 ng/L) in 1 L samples, a method for large volumes using XAD-7 resin was developed that allows detection to 0.01 ng/L. This method was applied to Lake Superior samples which produced good agreement for C6-C9 PFCAs between regular analysis (GC-MS) and the XAD-7 followed by GC-MS analysis. PMID- 17120573 TI - Investigation of novel incineration technology for hospital waste. AB - Conventional incineration systems for hospital waste (HW) emit large amounts of particulate matter (PM) and heavy metals, as well as dioxins, due to the large excess air ratio. Additionally, the final process residues--bottom and fly ashes containing high levels of heavy metals and dioxins--also constitute a serious environmental problem. These issues faced by HW incineration processes are very similar to those confronted by conventional municipal solid waste (MSW) incinerators. In our previous work, we developed a novel technology integrating drying, pyrolysis, gasification, combustion, and ash vitrification (DPGCV) in one step, which successfully solved these issues in MSW incineration. In this study, many experiments are carried out to investigate the feasibility of employing the DPGCV technology to solve the issues faced by HW incineration processes, although there was no MSW incinerator used as a HW incinerator till now. Experiments were conducted in an industrial HW incineration plant with a capacity of 24 tons per day (TPD), located in Zhenzhou, Henan Province. Results illustrated that this DPGCV technology successfully solved these issues as confronted by the conventional HW incinerators and achieved the expected results for HW incineration as it did for MSW incineration. The outstanding performance of this DPGCV technology is due to the fact that the primary chamber acted as both gasifier for organic matter and vitrifying reactor for ashes, and the secondary chamber acted as a gas combustor. PMID- 17120574 TI - Coagulation and electrocoagulation of wastes polluted with dyes. AB - Dyes are common pollutants in a large variety of industrial wastewaters, and the treatment of these wastes by coagulation has been extensively studied in the literature. This work is focused on the comparison of the efficiencies of the chemical and the electrochemical coagulation processes with hydrolyzing aluminum salts, and it tries to determine the similarities or differences that exist between the two coagulation processes. To do this, Eriochrome Black T solutions were used as a model of dye-polluted wastewater, and experiments of both coagulation technologies were planned to meet the same operation conditions. The pH, the aluminum concentration, the type of electrolyte, and the mode of dosing of aluminum were found to influence the process. Moreover, the speciation of aluminum was found to be the key parameter to explain the results, in terms of the mechanisms previously proposed in the literature for dissolved organic matter coagulation. PMID- 17120575 TI - Verifying three types of methane fluxes from soils by testing the performance of a novel mobile photoacoustic method versus a well-established gas chromatographic one. AB - The performance of a novel portable, tunable diode laser, resonant photoacoustic (TDL-PA) analyzer developed for field measurements of CH4 was compared to a commonly applied offline gas chromatographic (GC) method. This comparative studywas realized under normal field conditions parallel to long-term weekly GC monitoring of four different soil types with very different methane budgets. The method used for gas-exchange measurements was the well-known closed-chamber technique. The TDL-PA analyzer detects methane at 1650.957 nm [R (5) line of the 2v3 band], guaranteeing high precision without the need for correction procedures. The two techniques correlated well (R2 = 0.988) over the entire concentration range (0.15-33 ppmv CH4) tested at highly varying flux rates between -30 and -12 ppbv CH4 min(-1) for uptakes and between 2.5 and 362 ppbv CH4 min(-1) for emissions. The two analyzers proved to be interchangeable, leaving the online advantages to the TDL-PA. A suitable CH4 online GC solution for chamber measurement is not available as a portable system. Additionally, the data sampling rate of 2 Hz enables a direct coupling to other infrared gas analyzers with the high time resolution commonly required to determine plant CO2 assimilation rates or soil respiration rates. PMID- 17120576 TI - Effects of carbonate precipitates on long-term performance of granular iron for reductive dechlorination of TCE. AB - Long-term column experiments were conducted to evaluate the effects of secondary carbonate minerals on permeability and reactivity of commercial granular iron treating trichloroethene (TCE). The results showed that carbonate precipitates caused a decrease in reactivity of the iron, and spatially and temporally varying reactivity loss resulted in migration of mineral precipitation fronts, as well as profiles of TCE, pH, alkalinity, calcium, and dissolved iron. In the columns receiving solutions of dissolved calcium carbonate, porosity gradually decreased in proportion to the source concentrations, as carbonate minerals accumulated. However, the rate of porosity loss slowed over time because of the declining reactivity of the iron. Thus, secondary minerals are not likely to accumulate to the extent that there is a substantial reduction in hydraulic conductivity. The reactivity of the iron was found to decrease as an exponential function of the carbonate mineral volume fraction. This changing reactivity of iron should be incorporated into predictive models for improved designs of iron permeable reactive barriers (PRBs). PMID- 17120577 TI - Use of caustic magnesia to remove cadmium, nickel, and cobalt from water in passive treatment systems: column experiments. AB - In the present study caustic magnesia obtained from calcination of magnesium carbonate was tested in column experiments as an alternative material for passive remediation systems to remove divalent metals. Caustic magnesia reacts with water to form magnesium hydroxide, which dissolves increasing the pH to values higher than 8.5. At these pH values, cadmium is precipitated as otavite and to a minor amount as a hydroxide. Cobalt and nickel are precipitated as hydroxides which form isostructural solids with brucite. Thus, metal concentrations as high as 75 mg/L in the inflowing water are depleted to values below 10 microg/L. Magnesia dissolution is sufficiently fast to treat flows as high as 0.5 m3/m2 x day. For reactive grain size of 2-4 mm, the column efficiency ends due to coating of the grains by precipitates, especially when iron and aluminum are present in the solution. PMID- 17120578 TI - Role of moisture in adsorption, photocatalytic oxidation, and reemission of elemental mercury on a SiO2-TiO2 nanocomposite. AB - A novel silica-titania (SiO2-TiO2) nanocomposite has been developed to effectively capture elemental mercury (Hg0) under UV irradiation. Moisture has been reported to have an important impact on this nanocomposite's performance. In this work, the role of moisture on Hg0 removal and reemission as well as the corresponding mechanisms was investigated. Hg0 removal experiments were carried out in a fixed-bed reactor at 65 degrees C using air as the carrier gas. Without UV irradiation, Hg0 adsorption was found to be insignificant, but it could be enhanced by the photocatalytic oxidation product, mercuric oxide (HgO), possibly due to the high affinity between HgO and Hg0. Under dry conditions 95% of Hg0 can be removed; however, increased humidity levels remarkably suppress both Hg0 adsorption and photocatalytic oxidation. Introducing water vapor can also result in significant reemission of captured Hg0 from the nanocomposite, which may be ascribed to the repellant effect of water vapor adsorbed on the superhydrophilic TiO2 surface. Exposure to UV light was found either to prohibit Hg0 reemission when photocatalytic oxidation of reemitted Hg0 prevailed or to promote Hg0 reemission when photocatalytic reduction of HgO to Hg0 dominated later on. The results indicate that minimization of Hg0 reemission can be achieved by appropriate application of UV irradiation. PMID- 17120579 TI - Enrichment, performance, and microbial diversity of a thermophilic mediatorless microbial fuel cell. AB - A thermophilic mediatorless microbial fuel cell (ML-MFC) was developed for continuous electricity production while treating artificial wastewater concurrently. A maximum power density of 1030 +/- 340 mW/m2 was generated continuously at 55 degrees C with an anode retention time of 27 min (11 mL h(-1)) and continuous pumping of air-saturated phosphate buffer into the cathode compartment at the retention time of 0.7 min (450 mL h(-1)). Meanwhile, about 80% of the electrons available from acetate oxidation were recovered as current. Denaturing gradient gel electrophoresis (DGGE) and direct 16S-rRNA gene analysis revealed that the bacterial diversity in this ML-MFC system was lower than the inoculum. Direct 16S rDNA analysis showed that the dominant bacteria representing 57.8% of total population in anode compartment was phylogenetically very closely related to an uncultured clone, clone E4. Two sheets of graphite used as the anode showed different dominant bacterial population. For the first time, it is shown that thermophilic electrochemically active bacteria can be enriched to concurrently generate electricity and treat artificial wastewater in a thermophilic ML-MFC. PMID- 17120580 TI - Complete combustion of methane over indium tin oxides catalysts. AB - Indium tin oxide (ITO) catalysts with different In/Sn ratios have been prepared by the co-precipitation method. The catalysts were evaluated for methane combustion at different temperatures (673-873 K) with a space velocity of 30 000 h(-1). The results showed that methane could be completely oxidized at 873 K with ITO catalysts. Doping an appropriate amount of tin into In2O3 could greatly improve its activity, while the performances of Indium-doped tin oxides were worse than that of SnO2. A significant improvement of the activity was obtained on the catalyst In8Sn2, which contains 80 wt. % of indium oxide and 20 wt. % of tin oxide. Crystal defection and the amount of oxygen vacancy caused by doping were the main factors that would affect catalytic activity of ITO catalysts. The catalytic activity is strongly inhibited by the presence of a large amount of water vapor at the entire temperature range, while only the activity at low temperature (under 823 K) decreased in the presence of sulfur dioxide. By doping Sn into In2O3, its tolerance to SO2 could be enhanced due to the higher resistance of SnO2. PMID- 17120581 TI - Outside-in trimming of humic substances during ozonation in a membrane contactor. AB - This paper addresses the change of molecular size distribution of humic substances (HS) during ozonation in a membrane contactor. It focuses on the characterization and identification of some small ozonation products. The membrane contactor setup allows very precise control of ozone transfer into the solution as well as precise sampling of the products in time. The molecular size distribution was followed by gel permeation chromatography (GPC). Characterization and identification of small ozonation products was performed by membrane nanofiltration and high-performance liquid chromatography (HPLC). Measurements on molecular size distribution indicate that during the ozonation process the size of the HS molecules decreases slowly and only small highly oxidated compounds are being split off the larger molecules. Pyruvic acid, formic acid, methylglyoxal, and acetaldehyde could be identified by substantial peaks. Glyoxilic acid and glyoxal were identified to a lesser extent. This suggests that HS molecules consist of a relatively stable backbone network structure and that the HS molecule degrades according to an outside-in trimming mechanism. PMID- 17120582 TI - Long-term performance of high-rate anaerobic reactors for the treatment of oily wastewater. AB - Complex oily wastewater from a food industry was treated in three different UASB reactors at different operating conditions. Although all three systems achieved fat, oil, and grease (FOG) and COD removal efficiencies above 80% at an organic loading of 3 kg COD/m3 x d, system performance deteriorated sharply at higher loading rates, and the presence of high FOG caused a severe sludge flotation resulting in failure. Initially, FOG accumulated onto the biomass which led to sludge flotation and washout of biomass. The loss of sludge in the bed increased the FOG loading to the biomass and failure ensued. Contrary to previous findings, accumulation of FOG rather than influent FOG concentrations or volumetric FOG loading rate was the most importantfactor governing the high-rate anaerobic reactor performance. The critical accumulated FOG loading was identified as 1.04 +/- 0.13 g FOG/g VSS for all three reactors. Furthermore, FOG accumulation onto the biomass was identified mainly as palmitic acid (>60%) whereas the feed LCFA contained only 30% of palmitic acid and 50% of oleic acid. PMID- 17120583 TI - Stainless steels can be cathodically protected using energy stored at the marine sediment/seawater interface. AB - Laboratory-scale experiments were performed in which the corrosion protection of stainless steels in seawater was afforded by cathodic protection. The method was implemented for the first time using the potential difference at the marine sediment/seawater interface as the only source of electric power. Graphite electrodes buried in marine sediment, developing a potential of -0.45 V versus a saturated calomel electrode (SCE), were used as anodes to cathodically polarize UNS S30403 stainless steel coupons that were exposed to seawater. The cathodic protection system was operated with low polarization of stainless steel, typically to -0.2 V (vs SCE) and was found to properly prevent material failure even in the presence of a well-developed biofilm. With voltammetry, the protection current was found to be related to the oxidation of reduced sulfur compounds in the sediments. Results demonstrate that this inexpensive and environmentally friendly method can, so far, extend the service life of stainless steels in seawater. PMID- 17120584 TI - Development of the GENIPOL European flounder (Platichthys flesus) microarray and determination of temporal transcriptional responses to cadmium at low dose. AB - We have constructed a high density, 13 270-clone cDNA array for the sentinel fish species European flounder (Platichthys flesus), combining clones from suppressive subtractive hybridization and a liver cDNA library; DNA sequences of 5211 clones were determined. Fish were treated by single intraperitoneal injection with 50 micrograms cadmium chloride per kilogram body weight, a dose relevant to environmental exposures, and hepatic gene expression changes were determined at 1, 2, 4, 8, and 16 days postinjection in comparison to saline-treated controls. Gene expression responses were confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR). Blast2GO gene ontology analysis highlighted a general induction of the unfolded protein response, response to oxidative stress, protein synthesis, transport, and degradation pathways, while apoptosis, cell cycle, cytoskeleton, and cytokine genes were also affected. Transcript levels of cytochrome P450 1A (CYP1A) were repressed and vitellogenin altered, real-time PCR showed induction of metallothionein. We thus describe the establishment of a useful resource for ecotoxicogenomics and the determination of the temporal molecular responses to cadmium, a prototypical heavy metal pollutant. PMID- 17120585 TI - Assessing effects of metal mining effluent on fathead minnow (Pimephales promelas) reproduction in a trophic-transfer exposure system. AB - Assessment of effects of metal mine effluent (MME) on aquatic organisms in lab based settings predominantly evaluates contaminant transfer through the water only with little emphasis on food-borne exposure. The effects of MME on fathead minnow (Pimephales promelas) (FHM) have been reported downstream of metal mine discharges in the Junction Creek system, Sudbury, ON, but to date, no study has investigated the significance of trophic transfer in this system. Our objective was to develop a self-sustaining trophic-transfer bioassay, using Chironomus tentans and FHM, that allowed assessment of the effects of not only water-borne (FHM-only) but also food- and water-borne (trophic-transfer) exposure to MME on FHM reproduction. Reproductive performance of FHM was assessed for 21 days under controlled laboratory conditions to obtain baseline data of various endpoints, including egg production and hatching success. Exposure to 45% (v/v) Copper Cliff mine effluent (CCME) and control treatments for both systems was then conducted for a further 21 days. It was evident that reproductive output in both the water only and the trophic-transfer system was reduced compared to controls. It was only in the trophic-transfer system that a significant reduction in larval hatching and an increase in deformities occurred after exposure to CCME. This would suggest that contaminated food was a route of exposure causing effects on larval survival. PMID- 17120586 TI - First derivation of predicted-no-effect values for freshwater and terrestrial ecosystems exposed to radioactive substances. AB - The FASSET Radiation Effects Database (FRED) constitutes a unique structured resource of the biological effects of ionizing radiation on non-human species mainly from temperate ecosystems, encompassing 26,000 primary data entries. Quality-assessed data were extracted from FRED and dose-effect relationships were constructed to provide estimates of ED50 and EDR10. These estimates are Doses (or Dose Rates) related to the percent change in the average level of the endpoint for a particular effect (50% or 10% for acute or chronic exposure regimes, respectively). Acute and chronic Species Sensitivity Distributions (SSDs) were built on the basis of these data sets, and the Assessment Factor Method (AFM) was applied when data were too scarce. The Hazardous Dose corresponding to 5% of species acutely affected at the 50% effect level varied from 1 to 5.5 Gy according to the ecosystem. For chronic gamma external irradiation exposure, no effect values varied from 10 microGy/h for freshwaters through application of the AFM to 67 microGy/h for terrestrial ecosystems, corresponding to the 5th percentile of the non-weighted SSD (vs 229 microGy/h when trophic weights are applied). These values are higher by ca. x50 to x100 than the upper bound of natural background, and lower than dose rates triggering effects at individual levels on contaminated sites. PMID- 17120587 TI - Larval deformities associated with selenium accumulation in northern pike (Esox lucius) exposed to metal mining effluent. AB - The objective of this study was to investigate selenium toxicosis in larval northern pike (Esox lucius) originating from reproductively mature pike collected downstream of a uranium milling operation in northern Saskatchewan, Canada. Eggs were obtained from female pike collected from a reference site and three sites representing an exposure gradient (approximately 2, 10, and 15 km downstream of effluent discharge). Embryos were incubated following a two-way (crossover) analysis of variance experimental design that allowed discrimination between effects due to maternal transfer to eggs and effects due to site water exposure in the developing embryos. The major finding of this study was a significant increase in the frequencies of individual deformities (skeletal curvatures, craniofacial deformities, and fin deformities) and edema in fry originating from high and medium exposure site females (mean selenium concentrations of 48.23 and 31.28 microg/g egg dry weight and 38.27 and 16.58 microg/g muscle dry weight, respectively) compared to reference site females. Selenium concentrations resulting in a 20% increase in total deformities above background levels (EC20S) were 33.55 and 21.54 micro/g dry weight in eggs and muscle, respectively. Mathematical conversion of the egg- and muscle-derived relationships to whole body selenium levels resulted in similar EC20S of 15.56 and 17.72 microg/g dry weight, respectively. These relationships between tissue selenium levels and larval deformities suggest that northern pike are within the same range of sensitivity to selenium as the majority of warm water (e.g., centrarchids and cyprinids) and cold water (e.g., salmonids) fish species studied to date. PMID- 17120588 TI - Biomarkers of PAH exposure in an intertidal fish species from Prince William Sound, Alaska: 2004-2005. AB - Polycyclic aromatic hydrocarbon (PAH) exposure biomarkers were measured in high cockscomb prickleback (Anoplarchus purpurescens) fish collected from both previously oiled and unoiled shore in Prince William Sound (PWS), Alaska, to test the hypothesis that fish living in the nearshore environment of the sound were no longer being exposed to PAH from the Exxon Valdez oil spill. Pricklebacks spend their entire lives in the intertidal zone of rocky shores with short-term movements during feeding and breeding restricted to an area of about 15 meters in diameter. Fish were assayed for the PAH exposure biomarkers, bile fluorescent aromatic compounds (FAC), and liver ethoxyresorufin O-deethylase (EROD) activity (a measure of cytochrome P450 1A (CYP1A) monooxygenase activity). Bile FAC concentrations and EROD activities were low and not significantly different in fish from previously oiled and unoiled sites. The similar low EROD activity and bile FAC concentrations in fish from oiled and unoiled shores, supports the hypothesis that these low-level biomarker responses were not caused by exposure of the fish to residues of the spilled oil. PMID- 17120589 TI - A review of health related quality of life in systemic lupus erythematosus. AB - Health-related quality of life (HRQoL) is an important outcome measure in patients with systemic lupus erythematosus (SLE). A review was undertaken of the literature relating to HRQoL in SLE. MEDLINE, EMBASE, CINAHL, Allied and Complimentary Medicine were searched to locate full papers in the English language reporting on HRQoL in adult SLE patients published between 1990 and 2005. In total 53 papers were included and the review was subdivided into: 1) description of HRQoL in SLE patients; 2) HRQoL and disease activity and/or damage; 3) the impact of other variables on HRQoL; and 4) HRQoL measures used in clinical trials in SLE patients. The findings were as follows: HRQoL is reduced in SLE patients; HRQoL is not correlated to disease activity or damage; age appears to have a negative impact on HRQoL especially physical health but the effect of disease duration is unclear; other potentially modifiable variables such as fatigue and psychosocial factors impact on HRQoL in a complex manner; and HRQoL measures which are sensitive to change should be an essential outcome measure in all clinical trials on SLE patients. PMID- 17120590 TI - Increased aortic pulse wave velocity in middle aged women with systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a connective tissue disease where inflammatory activity affects several organ systems. An increased risk of cardiovascular disease has been identified in these patients, even after correction for traditional risk factors. The aim of the present study was to evaluate arterial stiffness and central hemodynamics in women with SLE in comparison to controls. Arterial tonometry was used to measure aortic (carotid femoral) and arm (carotid-radial) pulse wave velocity (PWV), reflected pressure waves, and aortic augmentation index (AIx) in 27 women with SLE (52 to 68 years) and 27 controls. Aortic PWV was higher in women with SLE than controls, 9.8 m/s versus 8.2 m/s (P < 0.01), after correction for mean arterial pressure and body mass index, 9.5 m/s versus 8.5 m/s (P < 0.05). Other parameters were similar, arm PWV, 8.4 versus 8.5 m/s, AIx 34 versus 33% and calculated central aortic pulse pressure 48 versus 43 mmHg, in SLE and controls, respectively (NS). Aortic PWV was positively associated to C-reactive protein (CRP) and complement factor 3 (C3). Women with SLE have increased stiffness of their elastic central arteries. This may be one factor contributing to the increased cardiovascular risk seen in this cohort. PMID- 17120591 TI - Neuropsychiatric manifestations in pediatric systemic lupus erythematosus: a 20 year study. AB - The objective of this study was to investigate the manifestations, treatment and outcome of neuropsychiatric (NP) involvement in pediatric systemic lupus erythematosus (SLE) patients. The charts of 185 pediatric patients with SLE diagnosed between 1985 and 2005 in a tertiary referral hospital were retrospectively reviewed. NPSLE were defined using the American College of Rheumatology NPSLE case definitions. NPSLE developed in 34.6% (64/185) of the patients. The mean onset age was 15.2 years. Fourteen patients (21.9%) had NP manifestations on initial diagnosis of SLE. The median duration from the onset of SLE to NP manifestation was 11 months. The most frequent NP manifestations were seizure disorder (84.4%), ischemic stroke (28.1%) and psychosis (21.9%). However, the prevalence of manifestations of NPSLE might be underestimated by the retrospective design of our study. Higher mean C3/C4 levels, less percentage of anti-dsDNA antibodies elevation and higher percentage of elevated anticardiolipin antibodies were observed in NPSLE events than in non-NPSLE events (P < 0.05). The mortality rate of NPSLE patients decreased from 52.2% in 1985-1994 cohort to 27.8% in 1995-2005 cohort. In the past 10 years, the leading cause of death in NPSLE patients was NPSLE itself. NPSLE is common in pediatric SLE patients. It has diverse manifestations and a high mortality. PMID- 17120592 TI - Study of a functional polymorphism in the p53 gene in systemic lupus erythematosus: lack of replication in a Spanish population. AB - The aim of this study was to assess the possible association between the p53 suppressor gene codon 72 polymorphism and systemic lupus erythematosus (SLE). Our study population consisted of 513 SLE patients and 567 healthy controls. All the individuals were of Spanish Caucasian origin. Genotyping of the p53 codon 72 polymorphism was performed by allele-specific PCR. No statistically significant differences were observed between SLE patients and healthy controls when p53 codon 72 genotype and allele frequencies were compared. In addition, no evidence for association with clinical subfeatures of SLE was found. In conclusion, the p53 codon 72 polymorphism associated with SLE in a Korean population does not appear to play a major role in the susceptibility or severity of SLE in the Spanish population. PMID- 17120593 TI - Geographical clustering of mortality from systemic lupus erythematosus in the United States: contributions of poverty, Hispanic ethnicity and solar radiation. AB - The objective was to investigate whether spatial variation in poverty, Hispanic ethnicity, and solar radiation explains the strong pattern of geographical clustering of mortality from systemic lupus erythematosus (SLE) in the United States. SLE mortality counts for women and men of black and white race in US counties, 1979-1998, were obtained from the National Center for Health Statistics. County-level poverty rates and proportions of Hispanic residents were drawn from the 1990 national census. The annual cumulative level of ambient ultraviolet 'B' radiation (UVB) was estimated for each county according to latitude, longitude, and elevation. Maps for the full study population and for sex - and race-specific subpopulations showed that the national pattern of geographical variation in SLE mortality primarily reflected the experience of white women. Formal spatial analysis of the data for white women identified 10 statistically significant, multi-county clusters--four with elevated and six with reduced SLE mortality rates. Multivariate regression modeling established that higher levels of poverty, Hispanic ethnicity, and UVB were each associated with elevated local rates of SLE mortality among white women. Statistical adjustment via the regression model was used to remove effects of these factors on local rates. In a re-application of spatial analysis to the adjusted rates, four clusters 'disappeared'. In those clusters, poverty, Hispanic ethnicity and UVB had explained an average of 58.2% of the deviations between local and national SLE mortality rates. In six clusters (including three that disappeared with adjustment), Hispanic ethnicity explained a larger percentage of the deviations between local and national rates than either poverty or UVB. In multivariate models based on data for black women and for men of both races, poverty and UVB had similar effects on SLE mortality rates to those observed among white women. However, Hispanic ethnicity was not a significant predictor of SLE mortality in any of these other groups. The contributions of poverty, Hispanic ethnicity and solar radiation to geographical variation of SLE mortality in the United States are substantial, but these factors do not fully account for the clustering phenomenon. PMID- 17120594 TI - Headache, Raynaud's syndrome and serotonin receptor agonists in systemic lupus erythematosus. AB - There are potential concerns regarding serotonin receptor agonists in SLE patients with migraine, particularly patients with concomitant Raynaud's syndrome. We estimated the prevalence of lupus-related headache and Raynaud's syndrome in the Montreal General Hospital SLE clinic cohort and evaluated the relationship between these two variables in multivariable logistic regression models, controlling for age, sex, race, SLE duration and the presence of lupus anticoagulant and antibodies to cardiolipin and beta2 glycoprotein I. We also assessed, through chart review in those individuals with both Raynaud's syndrome and migraine, a history of serotonin receptor agonist use, and any associated worsening vasospasm. Based on Systemic Lupus Activity Measure (SLAM) scores, the cumulative incidence of lupus-related headache in our sample (n = 391) was 46.1%; the prevalence of Raynaud's syndrome was 49.4%. The adjusted odds ratio (OR) for lupus-related headache and Raynaud's syndrome was 1.7 (95% CI 1.1, 2.5). In addition, there was a strong independent relationship between headache and anti beta2 glycoprotein I antibodies (adjusted OR 5.6 [95% CI 1.8, 17.0]). The data from our chart review suggest that careful use of serotonin receptor agonists in patients with both Raynaud's syndrome and migraines may be undertaken, although caution would necessitate that these agents not be used in individuals with very severe Raynaud's (eg, digital ulcerations, and so on). PMID- 17120595 TI - Traditional and non-traditional risk factors contribute to the development of accelerated atherosclerosis in patients with systemic lupus erythematosus. AB - To determine risk factors of accelerated atherosclerosis in patients with systemic lupus erythematosus (SLE), 72 patients with inactive disease and 36 age- and sex-matched controls were included. The intima-media thickness (IMT) of the common carotid artery was determined by ultrasound. Traditional risk factors and disease-related factors were recorded. Cardiovascular risk was estimated using SCORE (systematic coronary risk evaluation). Markers of inflammation, endothelial activation and vascular remodelling (matrix metalloproteinases (MMP-3, MMP-9) and tissue inhibitor of metalloproteinase- 1 (TIMP- 1)) were determined. IMT was increased in patients (0.67 mm+/-0.13 versus 0.61 mm+/-0.11, P < 0.05). Prevalence of hypertension (33% versus 6%, P < 0.001), SCORE (2.2 (1.7-4.2) versus 1.7 (1.3-2.1), P < 0.001), as well as parameters of inflammation (CRP 1.8 (0.6-5.8) mg/L versus 0.6 (0.2-1.0) mg/L, P < 0.001) and endothelial activation (VCAM-1 505 (389-683) ng/mL versus 374 (322-427) ng/mL, P < 0.001) and von Willebrand factor (138 (59-208)% versus 48 (24-92)%, P < 0.001), were increased in patients. Vascular remodelling was altered: MMP-3 and TIMP-1 were increased (18 (10-29) ng/mL versus 8 (5-11) ng/mL, P < 0.001, and 275 (216-352) ng/mL versus 230 (197-268) ng/mL, P < 0.001, respectively), and MMP-9 was decreased in SLE (266 (147-412) ng/mL versus 348 (226-530) ng/mL, P < 0.05). Univariate analyses revealed that in patients IMT was associated with age, systolic blood pressure, SCORE and disease duration. In multivariate analysis, age and SCORE were independent predictors of IMT. In conclusion, SLE patients have an increased IMT, which is associated with traditional risk factors. Non-traditional risk factors, such as endothelial activation, altered vascular remodelling and disease duration, might play an additional role. PMID- 17120596 TI - Successful treatment of cold agglutinin disease with anti-CD20 antibody (rituximab) in a patient with systemic lupus erythematosus. AB - Cold agglutinin disease (CAD) is a rare cause of anaemia in patients with systemic lupus erythematosus (SLE). CAD is usually refractory to glucocorticosteroid, and other immunosuppressive and/or cytotoxic therapies. We report the case of a 55-years old woman with SLE and CAD that did not respond to high-dose methylprednisolone, cyclosporin A, and double filtration plasma pheresis. Because several recent case reports and studies have indicated promising results of rituximab treatment for CAD and for SLE, rituximab was given weekly at 375 mg/m(2) in two doses. The rituximab was well tolerated, and there were no adverse effects. The hemolysis and SLE improved markedly, and the patient remained disease free eight months later. This is the first report of successful rituximab treatment of CAD in a patient with SLE. We conclude that rituximab is worth trying in such patients if they fail to respond to conventional treatment. PMID- 17120597 TI - Association of nodular regenerative hyperplasia of the liver with porto-pulmonary hypertension in a patient with systemic lupus erythematosus. AB - A 37-year old woman with systemic lupus erythematosus (SLE) complicated by pulmonary hypertension (PHT) was admitted to evaluate abnormal liver function. Radiological imaging study, including ultrasonography, computed tomography and magnetic resonance imaging and upper gastrointestinal endoscopy, revealed multiple hepatic nodules, hepatosplenomegaly and esophageal varices. Percutaneous needle liver biopsy showed non-cirrhotic hepatic nodules with hyperplastic hepatocytes surrounded by atrophic hepatocytes, confirming the diagnosis of nodular regenerative hyperplasia (NRH) associated with non-cirrhotic portal hypertension (PT). NRH of the liver is known to be a very rare hepatic manifestation in rheumatic diseases. This case shows the association of NRH with porto-pulmonary hypertension in SLE. PMID- 17120598 TI - Lupus nephritis in an Afro-Caribbean population: renal indices and clinical outcomes. AB - In an Afro-Caribbean population, 111 new cases of systemic lupus erythematosus were diagnosed in the 10-year period from January 1995. Fifty-three cases (48%) presented with or subsequently developed lupus nephritis (SLEN). We recorded clinical characteristics and treatment outcomes of SLEN. We retrospectively categorized patients into four groups based on presence or absence of proteinuria with or without renal impairment. Group 1 (n = 15, 28%) had normal renal function (creatinine clearance (CrCl) > 70 mL/minute) with urinary protein excretion (UPE) of 0.5-3.0 g/24 hour, group 2 (n = 7, 13%) had normal renal function with UPE > 3.0 g/24 hour, group 3 (n = 9, 17%) had renal impairment (CrCl < 70 mL/minute) with UPE of 0.5-3.0 g/24 hour and group 4 (n = 22, 42%) had renal impairment with UPE > 3.0 g/24 hour. Renal biopsies were performed in 15 patients (28%). The number of treated patients in-remission decreased across the groups, from 100% in group 1 and 71% in group 2, to 33% in group 3 and 32% in group 4 (Pr < 0.001). There were 12 deaths from renal causes: none in groups 1 and 2, two (22%) from group 3 and 10 (45%) from group 4 (Pr = 0.003). In resource-poor clinical settings with limited access to histopathological services, CrCl and UPE may be useful predictors of therapeutic response and clinical outcomes in SLEN. PMID- 17120599 TI - Malignancy prevalence in the first-degree relatives of persons with systemic lupus erythematous: a pilot study. PMID- 17120600 TI - What do lupus specialists believe about managing conventional cardiovascular risk factors in patients with systemic lupus erythematosus? PMID- 17120601 TI - Discontinuation rate and factors predictive of the use of hydroxychloroquine in LUMINA, a multiethnic US cohort (LUMINA XL). PMID- 17120602 TI - Communicating with patients when English is not their first language. PMID- 17120603 TI - Are we losing the battle against liver disease related mortality? PMID- 17120604 TI - HIV infection among heroin users and area of residence. AB - The aim of this study was to develop a hypothesis to explain the link between HIV prevalence and area of residence. The study was conducted in two parts using two existing data sources. In Part 1, the bloodborne viral test status and test results of a sample of clients attending treatment in December 2001 in two areas of Dublin, an inner city area (Dublin 8) and a suburban area (Dublin 24), were extracted from the Bloodborne Viral Status Dataset created by Grogan. In Part 2 the characteristics of heroin users seeking treatment for the first time at treatment services in their respective areas of residence, Dublin 8 or Dublin 24, between 1997 and 2000 were examined, using data from the National Drug Treatment Reporting System. A higher proportion of heroin users in Dublin 8 had HIV and hepatitis C than did their counterparts in Dublin 24. The analysis suggests that heroin users in Dublin 8 were more likely both to have ever used cocaine and to have used heroin daily, than were those who lived in Dublin 24. Also, a higher proportion of injectors living in Dublin 8 used heroin and cocaine concurrently than did their counterparts in Dublin 24. In both samples, heroin users who lived in Dublin 8 were older than those who lived in Dublin 24. The findings led to a hypothesis:'The risk of acquiring HIV is associated with area of residence and may be linked to cocaine use. PMID- 17120605 TI - Vulval cancer, 1992-2002 audit--multidisciplinary care and prognostic factors. AB - Vulval carcinoma is becoming increasingly common. Thirty-four cases of vulval carcinoma were treated from 01/01/1992-31/12/2002. The mean age was 67, range (18 90). The presenting complaints were "a lump" (76%)(25/33), "itch" (49%)(16/33), "discomfort" (30%)(10/33) and postmenopausal bleeding (21%)(7/33). Most patients presented with stage 1 or 2 disease (73%) (n = 24/33). The majority (97%) (32/33) underwent surgical treatment. Five-year survival was 61% (17/28), (disease-free survival 76% (13/17)). There were 12 cases of local/regional recurrence. Survival rates deteriorated with stage of disease. Lymph-node results, lowered survival from 79% (11/14), if negative, to 17% (1/6) if positive. Age >70 reduced survival from 69% (11/16) to 50% (6/12). We conclude that age, the stage of disease, and lymph-node status were important prognostic factors. The favourable outcomes reflect muItidisciplinary care--combining clinical examinations with regular home contact with specialist nurses, by telephone. PMID- 17120606 TI - Consensus on diabetes management in primary care. AB - Diabetes management in Irish Primary Care is in the early stages of development. This study used a Delphi technique with a panel of experts working in diabetes in primary care in order to reach a consensus on its future development. There was strong agreement on the need for multidisciplinary professional advocacy to improve services for people with diabetes. The panel indicated a preference for the development of shared models of care and emphasised the importance of research, audit and professional education. The results of this study can be used by health planners to develop services in a way that reflects the experience and opinion of those working as experts in the field of primary care diabetes. PMID- 17120607 TI - Anxiety and performance in elite non-professional athletes. AB - Anxiety is one of the main motivators with regards to performance of individuals in any given task, including sporting endeavours. Our study sought to assess state anxiety levels in elite non-professional sportsmen, and to investigate if anxiety correlated with sporting performance, the IDA-Q (irritability, depression & anxiety questionnaire) was used to assess 3 mental state variables in an inter county hurling team as well as a matched non-sporting control group, and performance was judged by completion of a standard task in 2 different settings: a non-pressurised one and a highly pressurised setting. Subjects had significantly higher anxiety scores on the IDA-Q than the controls (p = 0.019). There were no significant differences and controls in the depression and irritability scales. There was a significantly negative correlation between anxiety scores and performance on the IDA-Q; spearman r = -0.57. High anxiety levels impair sporting performance. PMID- 17120608 TI - Screening for patent ductus arteriosus. AB - The aim of our study was to look at the amount of training required to accurately assess ductal patency in very low birth weight infants and to show that this service could be provided by non-cardiologists. Training consisted of an introductory echocardiography course, a CD package, and a total of 3 hours of teaching with a paediatric cardiologist. All of this was accomplished within the 3 months preceding our study. All newborn infants less than 1500 grams or less than 30 weeks gestation had an echocardiogram performed in the first 48 hours of life. A prediction was then made based on the echocardiogram findings as to whether the arterial duct would become a clinically significant one. We based our prediction on established echocardiography criteria for the prediction of ductal patency, namely a ductal diameter of greater than 1.5 millimetres, a left atrial to-aortic ratio of greater than 1.4, and an abnormal pattern of flow in the distal aorta. In total, 20 very low birth weight infants were screened from March 2005 to June 2005. Fifteen infants did not develop any signs of an arterial duct and this was predicted correctly in 13 cases. Of the 5 infants who developed a clinically apparent duct 4 were predicted correctly, giving us a sensitivity and specificity of 80% and 87%. Our study shows that it is possible to predict ductal patency without extensive echocardiography training. It also shows us that a neonatology based screening programme for pre-symptomatic arterial ducts in very low birth weight infants would be feasible. PMID- 17120609 TI - Successful ex utero intrapartum treatment (EXIT) procedure for congenital high airway obstruction syndrome (CHAOS) owing to a large oropharyngeal teratoma. AB - Large fetal neck masses can present a major challenge to securing an airway at birth, with associated risks of hypoxia, brain injury and death. The authors report a case of a giant oropharyngeal teratoma diagnosed in a fetus of 19 weeks gestation. The fetus was delivered by the ex utero intrapartum treatment procedure allowing sufficient time on placental support for bronchoscopy and tracheostomy to secure the airway. A multidisciplinary team approach combined with an accurate prenatal diagnosis obtained through fetal ultrasound was the key to a successful outcome. Unfortunately due to the large size of tumour and intracranial extension, the lesion was unresectable and the baby died 6 days after birth. PMID- 17120610 TI - Hypercalcaemia in a case of florid pulmonary tuberculosis. PMID- 17120611 TI - Should cervical smears be taken at a postnatal visit? PMID- 17120612 TI - Clinical guidelines: indicators of rising or falling standards in healthcare delivery? PMID- 17120613 TI - Is pre conceptual advice effective in improving use of pre conceptual folic acid? PMID- 17120614 TI - Casualty overcrowding and increased mortality; is it time to say no? PMID- 17120615 TI - Black tea polyphenols protect against 7,12-dimethylbenz[a]anthracene-induced hamster buccal pouch carcinogenesis. AB - Dietary chemoprevention has emerged as a cost-effective approach for cancer control. We evaluated the chemopreventive effects of black tea polyphenols (Polyphenon-B) administration during the preinitiation phase of 7,12 dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis. The expression of proliferating cell nuclear antigen (PCNA) in the buccal pouch and the concentration of lipid peroxides, protein carbonyl, and the antioxidant status in the buccal pouch, liver and erythrocytes were used as biomarkers of chemoprevention. All the hamsters painted with DMBA alone for 14 weeks developed buccal pouch carcinomas associated with increased expression of PCNA, diminished lipid and protein oxidation, and enhanced antioxidant status. In the liver and erythrocytes of tumor-bearing animals, enhanced oxidation of lipids and proteins was accompanied by compromised antioxidant defenses. Dietary administration of Polyphenon-B effectively suppressed DMBA-induced HBP carcinogenesis as revealed by decreased incidence of tumours and PCNA expression. In addition, Polyphenon-B modulated lipid and protein oxidation and enhanced the antioxidant status in the pouch, liver, and erythrocytes. We suggest that Polyphenon-B exerts its chemopreventive effects by inhibiting cell proliferation in the target tissue and modulating the oxidant-antioxidant status in the target as well as in host tissues. PMID- 17120617 TI - Chemotherapy influences inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) activity on 3D breast cancer cell line. AB - Multicellular tumor spheroids (MTS) are three-dimensional structural forms of tumors grown in vitro in the laboratory. In this study, the aim was to determine the regulation of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) expressions on MTS in response to treatment with the commonly used anti-cancer drugs Doxorubicin and Docetaxel. The spheroids were generated using the "liquid overlay" technique. The distribution of both iNOS and eNOS was detected using indirect immunohistochemistry, while the expression of both iNOS and eNOS was measured using Western blots. Additionally, S-phase analysis using 5-bromo-2'-deoxyuridine (BrdU) was done on the MTS after treatment with doxorubicin, docetaxel, and a combination of the two. The Griess method was used to measure nitric oxide (NO) production in the cells. An increase in iNOS immunoreactivity and a decrease in eNOS immunoreactivity were observed after doxorubicin treatment, when compared with the other groups. Furthermore, upregulation of iNOS and downregulation of eNOS were detected in doxorubicin treated cells using Western blotting. Insignificant iNOS expression was observed in all of the groups, and it was particularly low in the control and drug combination groups. NO production was also found to be significantly high after docetaxel treatment, and cell proliferation decreased after doxorubicin treatment. In conclusion, chemotherapy influences NOS activity differently with the presence of different drugs. The results with iNOS show that doxorubicin is a more effective drug than docetaxel, and a drug combination may play a helpful role in the suppression of tumorigenicity and cancer metastasis. Interestingly, eNOS expression increased after the addition of both docetaxel and the drug combination, and it was found to negatively correlate with the histological grade of the tumor. Therefore, analyzing the expression of both iNOS and eNOS might be very useful for targeting the treatment of breast carcinoma and obtaining better information on prognosis. PMID- 17120616 TI - Phosphorylation of ErbB4 on tyrosine 1056 is critical for ErbB4 coupling to inhibition of colony formation by human mammary cell lines. AB - In many studies, ErbB4 expression in breast tumor samples correlates with a favorable patient prognosis. Similarly, ErbB4 signaling is coupled to cellular differentiation and growth arrest in a variety of model systems. However, in some studies, ErbB4 expression in breast tumor samples correlates with poor outcome. Likewise, studies using some human mammary tumor cell lines suggest that ErbB4 is coupled to malignant phenotypes. Thus, the roles that ErbB4 plays in human breast cancer are still poorly defined. Here we demonstrate that a constitutively active ErbB4 mutant (ErbB4-Q646C) inhibits colony formation on plastic by two human mammary tumor cell lines (SKBR3 and MCF7) and by the MCF10A immortalized human mammary cell line, but does not inhibit colony formation by the MDA-MB-453 and T47D human mammary tumor cell lines. ErbB4 kinase activity is necessary for ErbB4 function and phosphorylation of ErbB4 Tyr1056 is necessary and appears to be sufficient for ErbB4 function. The inhibition of colony formation by MCF10A cells is accompanied by growth arrest but not cell death. These data suggest that ErbB4 behaves as a mammary tumor suppressor and that loss of ErbB4 coupling to growth arrest may be an important event in mammary tumorigenesis. PMID- 17120618 TI - Association of promoter hypermethylation of the RASSF1A gene with prognostic parameters in endometrial cancer. AB - RASSF1A is a tumor suppressor gene that is frequently hypermethylated in various human cancers. In the present study, we examined RASSF1A methylation status in 70 patients with endometrial cancer to search for correlations between the promoter hypermethylation of RASSF1A and the clinicopathologic parameters. Thirty-six of 70 endometrial cancers demonstrated hypermethylation of the RASSF1A promoter. Advanced stage disease (FIGO stage III, IV), lymph node involvement, and high grade (G3) are more frequent in patients with RASSF1A hypermethylation than in those without. We also observed a higher incidence of recurrences and lower disease-free survival (DFS) in patients with RASSF1A hypermethylation (77.8% and 97.0% at 5 years for methylated and unmethylated patients, respectively, p = 0.039). Our results suggest that RASSF1A hypermethylation might be a useful indicator of tumor aggressiveness in endometrial cancer patients. PMID- 17120619 TI - A review of the factors affecting the costs of bovine mastitis. AB - Mastitis is one of the most prevalent production diseases affecting the dairy cattle industry worldwide. Its occurrence is associated with direct and indirect losses and expenditures. When estimating the cost of mastitis to the dairy industry the cost of the control programmes must be added. The direct losses of mastitis are the only costs obvious to the farmer. The difference between the costs of mastitis on one side and the benefits of mastitis control on the other side will give us a picture of the economic efficacy of the mastitis control programme. Continuing education of the farmer is needed for better mastitis control programmes. This article is an attempt to review briefly all relevant factors included in the economics of bovine mastitis and to illustrate the authors' view of some of the costs. PMID- 17120620 TI - Seroprevalence of Babesia bovis and Babesia bigemina in cattle in the Soutpansberg region, Limpopo Province, South Africa, associated with changes in vector-tick populations. AB - A survey was conducted at 30 communal dip tanks and on 5 commercial farms in Limpopo Province, South Africa, during 1999 and 2000 to determine the seroprevalence of antibodies to Babesia bovis and Babesia bigemina. Cattle seropositive for B. bovis were found in 97% of the herds on communal land; the overall seroprevalence changed little between 1999 (63.3%) and 2000 (62.4%). All herds surveyed were infected with B. bigemina, and overall seroprevalence decreased significantly from 56.1% in 1999 to 49.3% in 2000. In herds on communal land in Sour Lowveld Bushveld, overall seroprevalence of B. bovis increased from 70% in 1999 to 80% in 2000, while seroprevalence of B. bigemina decreased from 70% in 1999 to 30% in 2000. This was possibly due to an influx of Rhipicephalus (Boophilus) microplus that occurred at the time. In commercially farmed herds the seroprevalence to B. bovis increased significantly from 19% in 1999 to 57.5% in 2000. All commercial herds in the survey tested positive to B. bigemina, with a seroprevalence of 48.3% in 1999 and 47.5% in 2000. During 1999, cattle in 60% of the dip tank/farm herds with only R. (B.) microplus present were approaching endemic stability to both B. bovis and B. bigemina. In 2000, 60% of the herds with only R. (B.) microplus present were approaching endemic stability for B. bovis, while only 45% were approaching endemic stability for B. bigemina. Those dip tanks/farms where only R. (B.) microplus was recorded had a significantly higher seroprevalence of B. bovis than those where both tick species were present. PMID- 17120621 TI - The use of a pour-on and spray dip containing Amitraz to control ticks (Acari: Ixodidae) on cattle. AB - Knockdown and persistence efficacies of a pour-on containing Amitraz 1% and Cypermethrin 1% and a spray dip containing 12.5% Amitraz were compared. Knock down and persistence efficacies of the Amitraz spray dip against Rhipicephalus (Boophilus) decoloratus and Amblyomma hebraeum were significantly higher for the duration of the trial than those of the pour-on. In the case of Rhipicephalus appediculatus and Rhipicephalus evertsi evertsi, efficacy was significantly higher on Days 2 and 5. Resistance testing prior to the start of the trial indicated that R. (B.) decoloratus was resistant to both Cypermethrin and Amitraz, yet in the case of the spray dip excellent efficacy results were obtained. It is thought that the cattle's scruffy winter coat may have hindered the spread of the pour-on, but that the thorough wetting and especially the higher concentration of active ingredient applied via the spray dip allowed this formulation to be effective. These results show that under certain conditions a spray dip containing 12.5% Amitraz may be more effective than a pour-on containing 1% Amitraz and 1% Cypermethrin, despite apparent in vitro resistance. PMID- 17120622 TI - A method for determining the extent of thermal burns in elephants. AB - A practical method was developed to assess the extent of burns suffered by elephants caught in bush fires. In developing this method, the surface areas of the different body parts of juvenile, subadult and adult elephants were first determined using standard equations, and then expressed as a percentage of the total body surface area. When viewed from a distance, the burnt proportion of all body segments is estimated, converted to percentages of total body surface area, and then summed to determine the extent of burns suffered. PMID- 17120623 TI - Telemedicine in veterinary practice. AB - Veterinary surgeons have a long tradition of consulting one another about problem cases and many have unwittingly practised telemedicine when discussing cases by telephone or by sending laboratory reports by telefax. Specific veterinary telemedicine applications have been in use since the early 1980s, but little research has been undertaken in this field. The Pubmed and CAB International databases were searched for the following Boolean logic-linked keywords; veterinary and telemedicine, veterinary and telecare, animal and telemedicine, animal and telecare and veterinary and e-mail and an additional search was made of the worldwide web, using Google Scholar. This returned 25 papers which were reviewed. Of these only 2 report research. Sixteen papers had no references and 1 author was associated with 13 papers. Several themes emerge in the papers reviewed. These include remarks about the use of telemedicine, the benefits that can and are derived from the use of telemedicine, areas of practice in which telemedicine is being used, ethical and legal issues around the practice of telemedicine, image standards required for telemedicine, the equipment that is required for the practice of telemedicine, advice on ways in which digital images can be obtained and educational aspects of telemedicine. These are discussed. Veterinary practice has lagged behind its human counterpart in producing research on the validity and efficacy of telemedicine. This is an important field which requires further research. PMID- 17120624 TI - Acellular dermal graft for repair of abdominal wall defects in rabbits. AB - Sixteen clinically healthy New Zealand white rabbits of either sex were divided into 2 equal groups (I and II) of 8 animals each. Under thiopental sodium (2.5%) anaesthesia a 2 x 3 cm full-thickness abdominal wall defect in the mid-ventral abdominal wall was created and repaired with an acellular dermal graft (ADG) in all the animals of group I (test group). In animals of group II (control group) a full-thickness linear midline abdominal muscular wall incision was made and repaired with a continuous suture pattern using 2-0 nylon. PMID- 17120625 TI - Confirmed Datura poisoning in a horse most probably due to D. ferox in contaminated tef hay. AB - Two out of a group of 23 mares exposed to tef hay contaminated with Datura ferox (and possibly D. stramonium) developed colic. The 1st animal was unresponsive to conservative treatment, underwent surgery for severe intestinal atony and had to be euthanased. The 2nd was less seriously affected, responded well to analgesics and made an uneventful recovery. This horse exhibited marked mydriasis on the first 2 days of being poisoned and showed protracted, milder mydriasis for a further 7 days. Scopolamine was chemically confirmed in urine from this horse for 3 days following the colic attack, while atropine could just be detected for 2 days. Scopolamine was also the main tropane alkaloid found in the contaminating plant material, confirming that this had most probably been a case of D. ferox poisoning. Although Datura intoxication of horses from contaminated hay was suspected previously, this is the 1st case where the intoxication could be confirmed by urine analysis for tropane alkaloids. Extraction and detection methods for atropine and scopolamine in urine are described employing enzymatic hydrolysis followed by liquid-liquid extraction and liquid chromatography tandem mass spectrometry (LC/MS/MS). PMID- 17120626 TI - Unusual manifestation of a concurrent demodectic and sarcoptic mange in a Zebu Friesian cross-bred heifer. AB - We describe a rare case of a concurrent demodectic and sarcoptic mange in a 2 year-old heifer in Khartoum, Sudan. The lesions were massive lumps of granulomatous tumour-like dermatitis with thick, nodular folds mainly covering the head, neck and shoulders. Histopathological examination of the lesions revealed the presence of both Demodex bovis and Sarcoptes scabiei. The animal died regardless of the anti-parasitic treatment it received. PMID- 17120627 TI - Significant climate change of extreme rainfall in Denmark. AB - During recent years, practitioners of urban drainage have complained that they felt that surcharging occurred more frequently. Therefore, a study was initiated focussing on the variations in extreme rainfall during the last two decades. In Denmark, a total of 41 rain gauges with a high resolution in time and volume have an observation period close to 20 years. The rainfall observed at these gauges was selected for this study. Three variables were analysed for statistically significant trends during the observation period: Maximum average 10 min intensity, maximum average 6h intensity and the total volume of individual rain events. For the 10 min maximum intensity there is a statistically significant trend towards more extreme and more frequently occurring rain storms. The trend is pronounced in the eastern part of the country and below statistically significant levels in the western part of the country. For the 6 h maximum intensity and total volume of events the trends are less pronounced. The findings are confirmed by comparison to physically based climate models and studies based on large regions. PMID- 17120628 TI - The effect of climate change on urban drainage: an evaluation based on regional climate model simulation. AB - That we are in a period of extraordinary rates of climate change is today evident. These climate changes are likely to impact local weather conditions with direct impacts on precipitation patterns and urban drainage. In recent years several studies have focused on revealing the nature, extent and consequences of climate change on urban drainage and urban runoff pollution issues. This study uses predictions from a regional climate model to look at the effects of climate change on extreme precipitation events. Results are presented in terms of point rainfall extremes. The analysis involves three steps: Firstly, hourly rainfall intensities from 16 point rain gauges are averaged to create a rain gauge equivalent intensity for a 25 x 25 km square corresponding to one grid cell in the climate model. Secondly, the differences between present and future in the climate model is used to project the hourly extreme statistics of the rain gauge surface into the future. Thirdly, the future extremes of the square surface area are downscaled to give point rainfall extremes of the future. The results and conclusions rely heavily on the regional model's suitability in describing extremes at timescales relevant to urban drainage. However, in spite of these uncertainties, and others raised in the discussion, the tendency is clear: extreme precipitation events effecting urban drainage and causing flooding will become more frequent as a result of climate change. PMID- 17120629 TI - On the issue of trend and noise in the estimation of extreme rainfall properties. AB - Up until today extreme rainfall properties are frequently applied in sewer design guidelines. Uncertainty in the estimation of such properties will hence directly influence the dimensions of sewers, structures and pumps. In this paper the issue of potential trend and noise in the estimation is investigated for 6 rain series ranging from 19 to 55 years in duration. Different to recent research that predicts a climate induced-increase in heavy precipitation no clear indication for such trend was found in the investigated historical rain series. Another important aspect is the length of the rain series that is required for the estimation of extreme rainfall properties and the associated uncertainty. The analysis indicates that at least a period of 10 years should be used for the estimation. But even so the possible deviation expressed in terms of the 90 percentile is in the order of 5 to 10% of the 'true value', defined as the value derived when the whole given series is used for the estimation. PMID- 17120630 TI - On the effect of spatial variances in historical rainfall time series to CSO performance evaluation. AB - Historical, high-resolution rain series are the backbone of modern combined sewer overflow (CSO) structure design. These rain series are the input to the computational estimation of the performance of the measures with respect to CSO pollution abatement. However, those historical precipitation measurements are available at only a few locations. Frequently rain series have to be used from gauging stations at a significant distance. In order to judge and to compensate for this influence an estimate between rain characteristics and combined sewer outflow (CSO) performance indicators would be useful. In this paper such correlations have been sought for a collection of 37 rain series covering large areas of Europe. It was found that the mean annual rain volume can explain most of the variances for the performance indicators Number of overflows and CSO volume. For explaining the spatial differences in the efficiency of the CSO structure another rain characteristic, i.e. the maximum event with a return period of one year, is to be used. PMID- 17120631 TI - From point to areal rainfall: linking the different approaches for the frequency characterisation of rainfalls in urban areas. AB - In urban water design and management, many hydrologic problems involve the frequency characterisation of rainfalls. Hydrologists are commonly asked to evaluate rainfall intensities for given recurrence frequencies or to indicate how rare an observed event is by estimating its return period. This study aims to improve the characterisation of rainfall hazard over a city by linking point to areal rainfall frequency analysis. We use a stochastic rainfall field generator based on the turning-bands method directly to assess areal rainfall distributions and to illustrate the link between different approaches. The simulating algorithm is applied to rainfall data from the city of Marseilles. The frequency analysis of simulated fields provides the elements to deal with the notions of return period and severity of observed storm events. The study concludes on the importance of a unified approach to assess rainfall better. PMID- 17120632 TI - Study of the water budget of streets: experimentation and modelling. AB - The study of two stretches of street during 38 months has been performed to analyze the hydrological behavior of streets during rain events. The results show that runoff coefficients are very variable and runoff losses may be important. In order to better understand this behavior, a physically based model has been used. This model, BiL, combines a porous media flow module with a surface runoff module. The lateral runoff transfer in the lateral gutter is approximated by the Muskingum model. Evaporation is simulated by an adaptation of the Penman method. A sensitivity study shows that the model is mainly sensitive to saturated hydraulic conductivity of the asphalt pavement and to the storage capacity. The comparison of simulated and observed data gives good results for the runoff outflow at a 3 minutes time step. Nevertheless, the simulation results are less encouraging for the runoff coefficient. This study of the water budget of two street stretches during a time period of 38 months indicates that the infiltration and evaporation represent between 20 and 30% of rain. PMID- 17120633 TI - Assessment of runoff contributing catchment areas in rainfall runoff modelling. AB - In numerical modelling of rainfall caused runoff in urban sewer systems an essential parameter is the hydrological reduction factor which defines the percentage of the impervious area contributing to the surface flow towards the sewer. As the hydrological processes during a rainfall are difficult to determine with significant precision the hydrological reduction factor is implemented to account all hydrological losses except the initial loss. This paper presents an inconsistency between calculations of the hydrological reduction factor, based on measurements of rainfall and runoff, and till now recommended literature values for residential areas. It is proven by comparing rainfall-runoff measurements from four different residential catchments that the literature values of the hydrological reduction factor are over-estimated for this type of catchment. In addition, different catchment descriptions are presented in order to investigate how the hydrological reduction factor depends on the level of detail regarding the catchment description. When applying a total survey of the catchment area, including all possible impervious surfaces, a hydrological reduction factor of approximately 0.5 for residential areas with mainly detached houses is recommended contrary to the literature recommended values of 0.7-0.9. PMID- 17120634 TI - From single-objective to multiple-objective multiple-rainfall events automatic calibration of urban storm water runoff models using genetic algorithms. AB - The calibration of storm water runoff models is a complex task. Early attempts focused on the choice of a performance criterion function that could capture all the facets of the problem into a single-objective framework. Subsequently, the awareness that a good calibration must necessarily take into account conflicting objectives led to the adoption of more sophisticated multi-objective approaches. Only recently, the focus has shifted towards effective ways of exploiting the mounting information provided by the availability of many sets of concurrent rainfall and flow measurements. This paper revisits through a case study the transition just elucidated: the calibration of a SWMM model applied to a catchment in Singapore is tackled through a single-objective, a multi-objective and a multi-objective multiple-event (MOME) paradigm respectively. A new approach to support the latter is presented herein. It consists in formulating the problem of model calibration as a multi-objective problem with m x r objective functions, where m and r are the number of performance criteria and rainfall events respectively, that must be optimized simultaneously. Results suggest that the new MOME framework performs significantly better than the others tested on the case study presented. PMID- 17120635 TI - Comparison of numerical and experimental simulations of a flood in a dense urban area. AB - Although various numerical methods were used to simulate real floods occurring in cities, the validation of the models was never accurate because of the lack of data about location and event description and about observation for validation. In order to check the capacities of our 2-dimensional shallow water equations model to simulate an urban flood, we then decided to simulate numerically an experimental event with well known characteristics and accurate flow measurements. The physical model presented in (Ishigaki et al, 2003) represents the flooding of the city center of Kyoto in Japan due to an overflow from the Kamo river. The 2-dimensional numerical simulation of this event was then set up and the experimental and computed data were compared. It appears that the event was calculated quite fairly in terms of flow depth and flow rates in the streets and in terms of timing. However, some discrepancies appear between the measurements and the numerical results, mostly due to some topographical local uncertainties and to the capacities of the equations to model the complex flows in the crossroads. PMID- 17120636 TI - Flooding flows in city crossroads: experiments and 1-D modelling. AB - This study focuses on the discharge distribution in an intersection of four channels, similar to a city crossroad. The channels and the intersection are all horizontal. Flow enters through two of the channels, and leaves through the other two. The flow is subcritical everywhere, and flow depths are controlled by vertical weirs at the exits of the outlet channels. The main variables that are measured are the flow rates in the four channels. When the weir heights in the outlet channels are the same, the ratio of flow rates in the outlet channels depends only on the ratio of flow rates in the inlet channels; if the outlet conditions are different, other parameters, such as the total flow rate also become important. The flow has also been simulated numerically using a solution of the 1-D Saint Venant equations, with a simple model to predict flow distribution in the intersection. A comparison with the experimental data shows that this model works well for the limited range of experimental conditions studied here. However, further work is needed on a wider range of conditions, closer to real conditions, before the model can be considered valid for practical applications. PMID- 17120637 TI - One-dimensional GIS-based model compared with a two-dimensional model in urban floods simulation. AB - A GIS-based one-dimensional flood simulation model is presented and applied to the centre of the city of Nimes (Gard, France), for mapping flow depths or velocities in the streets network. The geometry of the one-dimensional elements is derived from the Digital Elevation Model (DEM). The flow is routed from one element to the next using the kinematic wave approximation. At the crossroads, the flows in the downstream branches are computed using a conceptual scheme. This scheme was previously designed to fit Y-shaped pipes junctions, and has been modified here to fit X-shaped crossroads. The results were compared with the results of a two-dimensional hydrodynamic model based on the full shallow water equations. The comparison shows that good agreements can be found in the steepest streets of the study zone, but differences may be important in the other streets. Some reasons that can explain the differences between the two models are given and some research possibilities are proposed. PMID- 17120638 TI - Bottom shear stress in unsteady sewer flow. AB - The properties of unsteady open-channel turbulent flow were theoretically and experimentally investigated in a circular cross section channel with fixed sediment deposits. Velocity and turbulence distribution data were obtained using an ultrasonic velocity profiler (UVP). Different uniform flow conditions and triangular-shaped hydrographs were analysed. The hydrograph analysis revealed a dynamic wave behaviour, where the time lags of mean cross section velocity, friction velocity, discharge and flow depth were all evident. The bottom shear stress dynamic behaviour was estimated using four different approaches. Measurements of the velocity distribution in the inner region of the turbulent layer and of the Reynolds stress distribution in the turbulent flow provided the analysed data sets of the bottom shear stress. Furthermore, based on the Saint Venant equation, the bottom shear stress time behaviour was studied using both the kinematic and the dynamic flow principles. The dynamic values of the bottom shear stress were compared with those for the steady flow conditions. It is evident that bottom shear stress varies along the generated flood hydrograph and its variation is the function of the flow unsteadiness. Moreover, the kinematic flow principle is not an adequate type of approximation for presented flow conditions. PMID- 17120639 TI - Experimental analysis of the hydrass flushing gate and laboratory validation of flush propagation modelling. AB - Periodical cleansing operations prove necessary inside sewer systems in order to reduce hydraulic and environmental problems owing to the accumulation of deposits on the bottom of channels. For this objective, new effective hydraulic devices based on the scouring effects of flushing waves have been recently set up and adopted in many sewer systems. In this paper, the results of an experimental and numerical investigation on the hydraulic operation of the Hydrass flushing gate are reported. The experimental analysis has been carried out using a laboratory channel and a reduced scale model of the gate, in order to characterise the flushing waves generated by the device. The numerical analysis has been performed using a mathematical model specifically developed for the simulation of flushing waves inside sewer channels. The comparison of numerical results and experimental data has allowed evaluation of the applicability under unsteady flow conditions of the outflow relations determined for the Hydrass gate in a previous investigation under steady flow conditions. PMID- 17120640 TI - Long term monitoring of sewer sediment accumulation and flushing experiments in a man-entry sewer. AB - This paper presents continuous field experiments carried out during 4 years in a man-entry egg-shaped combined sewer in Lyon, France in order to contribute to the knowledge and the modelling of sediment accumulation and sediment removal by means of a Hydrass flushing gate. The 250 microm sediments are mainly mineral, and their physical and chemical characteristics appear as rather stable in time and space. Long-term sediment monitoring reveals: (i) a regular asymptotic increase of both the sediment mass and the slope of its longitudinal profile, (ii) a clear correlation between local sediment profile irregularities and sewer ancillaries but without significant influence on the global and long term accumulation. Simple sediment accumulation modelling shows: (i) a good suitability of a three parameters conceptual model to reproduce asymptotic sediment volume accumulation, and (ii) a good suitability of the Velikanov model to reproduce sediment profiles. Both models reproduce observations with an acceptable margin of uncertainty for operational management purposes but are very sensitive to input data and parameter values. The Hydrass flushing gate is efficient and it appeared that the mass of sediments moves downstream linearly with the number of flushes. PMID- 17120641 TI - Failure of sewage pumps: statistical modelling and impact assessment. AB - Sewage pumping stations are directly responsible for affecting performance, i.e. failing pumps may result in combined sewer overflows or flooding. However, failures of sewage pumps are not yet incorporated in sewer assessments due to lack of knowledge and data. This paper presents the analysis of pump failure data provided by two sewer management authorities in The Netherlands. Pump failures have been studied accounting for the nature of the failures, the operation and maintenance procedures of the management authority, the ageing of the pumps and the changes in the environment of pumps. The analysis shows that sewage pumps fail relatively often due to the composition of sewage and the discontinuous operation of the pumps. The interarrival time and the duration of failures are highly variable and independent of the pump type and the specific function of the pump. The results also indicate that the serviceability of sewer systems is significantly affected by failing pumps. As a consequence, part of the environmental damage due to CSOs (combined sewer overflows) can be avoided by improving maintenance of pumping stations. PMID- 17120642 TI - Recent findings on sinks for sulfide in gravity sewer networks. AB - Sulfide buildup in sewer networks is associated with several problems, including health impacts, corrosion of sewer structures and odor nuisance. In recent years, significant advances in the knowledge of the major processes governing sulfide buildup in sewer networks have been made. This paper summarizes this newly obtained knowledge and emphasizes important implications of the findings. Model simulations of the in-sewer processes important for the sulfur cycle showed that sulfide oxidation in the wetted biofilm is typically the most important sink for dissolved sulfide in gravity sewers. However, sulfide emission and thereby potential hydrogen sulfide buildup in the sewer atmosphere is of particular importance in sewers constructed with large diameter pipes, in sewers constructed with steep slopes and in sewers conveying low pH wastewater. Precipitation of metal sulfides is only important when the sulfide concentration in the wastewater is low; i.e. less than 1 g Sm(-3). PMID- 17120643 TI - Quantification of sewer leakage: a review. AB - One of the goals of the APUSS project (assessing infiltration and exfiltration on the performance of urban sewer systems) is to assess sewer exfiltration, in order to support cities and operators to define problem-oriented rehabilitation strategies. In this paper, we review various methods currently used for the estimation of exfiltration and discuss data needs and applicability for rehabilitation planning. Although each approach has its individual advantages and drawbacks, we identified pressure tests and tracer methods to have the highest potential for decision support in rehabilitation planning. With regard to future challenges (e.g., micropollutants) such methods might play a key role in integrated sewer management. PMID- 17120644 TI - The effectiveness of selected microbial and chemical indicators to detect sewer leakage impacts on urban groundwater quality. AB - Sewer and stormwater pipe leakage can lead to the degradation of urban groundwater quality. This groundwater may be subsequently used for public water supply and so the resulting water treatment and public health consequences can be serious. To understand the impact of sewer exfiltration on groundwater quality, suitable indicators need to be sampled and analysed for. This study examined potential sewer-derived inorganic and microbial parameters in the U.K. city of Doncaster. Sulphite reducing clostridia, faecal streptococci and boron were all detected in groundwater with reductions compared with sewer values ranging from 1 to 6 orders of magnitude for the former two, to 1 to 2 orders of magnitude decrease for boron. The correlation between these two different indicator types suggests that groundwater quality is being adversely affected by sewer leakage in the study area. The employment of several and varied indicators can better demonstrate the effect than use of single parameters. PMID- 17120645 TI - Statistical methods towards more efficient infiltration measurements. AB - A comprehensive knowledge about the infiltration situation in a catchment is required for operation and maintenance. Due to the high expenditures, an optimisation of necessary measurement campaigns is essential. Methods based on multivariate statistics were developed to improve the information yield of measurements by identifying appropriate gauge locations. The methods have a high degree of freedom against data needs. They were successfully tested on real and artificial data. For suitable catchments, it is estimated that the optimisation potential amounts up to 30% accuracy improvement compared to nonoptimised gauge distributions. Beside this, a correlation between independent reach parameters and dependent infiltration rates could be identified, which is not dominated by the groundwater head. PMID- 17120646 TI - Using decision analysis to determine optimal experimental design for monitoring sewer exfiltration with tracers. AB - The tracer methods developed to assess exfiltration from sewers in the European project APUSS (assessment of the performance of sewer systems) have a high degree of freedom with regard to the choice of tracer and the dosing strategy. These can lead to very different degrees of uncertainty in the measured exfiltration ratio. In this study, we demonstrate how to select an optimal experimental design using decision analysis, which accounts for this uncertainty and its associated costs. Although the results are site-specific, we can conclude generally that, when NaCl is used as the tracer, the accuracy of the exfiltration estimate is most sensitive to the amount of tracer used and the starting time of the experiment. PMID- 17120647 TI - Sampling for representative micropollutant loads in sewer systems. AB - Most commercially available auto-sampling devices do not support a continuous flow-proportional sampling mode, which would conceptually be the best for collecting representative composite samples. Instead different discrete sampling modes are available. Household chemicals can show considerable random short-term variations. With the anticorrosive benzotriazole, relating to a middle-frequent household activity, we show that, besides an accurate flow meter, mainly three factors are decisive for the representativeness of a substance's average load: the substance's load pattern, the sampling frequency and the length of the composite sample. When the sampling intervals are 10 minutes or longer, errors in the order of +/-40% (standard deviation) or more have to be accepted, if the substance of interest is contained in a low number of wastewater pulses (i.e., the level of household activity). This particularly holds true for specific pharmaceuticals e.g. carbamazepine. Ammonium would be less critical, because it relates to a larger number of sources in the same catchment. PMID- 17120648 TI - Quality of stormwater runoff from paved surfaces of two production sites. AB - In order to investigate stormwater pollutant loads associated with different anthropic activities and the related pollutant build-up and wash-off processes, two pilot sites have been equipped in the Liguria Region (Italy) for monitoring first flush water quality in a gas station and an auto dismantler facility. TSS, COD, HCtot and heavy metals in dissolved form (Zn, Pb, Cu, Ni, Cd, Cr) have been analyzed during the monitoring campaign (started in February 2004). Stormwater flow and quality data collected in both production sites confirm that EMC values are significantly higher than those observed in an urban site. In the auto dismantler site, the EMC values for TSS, COD and HC largely exceed the standard values (EC 91/271). Contrary to urban surface runoff, scarce correlation between TSS and COD concentrations is observed in runoff from both production sites. The occurrence and nature of the pollutant load connected to first flush flows is discussed by inspection of the M(V)-curves that are provided for all monitored water quality parameters. Significant first flush phenomenon is evidenced for TSS and HC, while such clear behavior doesn't emerge for heavy metals. Hydrologic and climatic characteristics (ADWP, rainfall intensity/depth) appear to scarcely affect the build-up and wash-off processes. PMID- 17120649 TI - Spatial variability of polycyclic aromatic hydrocarbon load of urban wet weather pollution in combined sewers. AB - In Paris, the OPUR research programme created an experimental on-site observatory of urban pollutant loads in combined sewer systems in order to characterise the dry and wet weather flows at different spatial scales. This article presents the first results on the spatial variability of the polycyclic aromatic hydrocarbon (PAH) load during wet weather flow (WWF). At the scale of a rain event, investigations revealed that (i) PAH concentrations were relatively homogenous whatever the spatial scale and were greater than those of the dry weather flow (DWF), (ii) PAH distributions between dissolved and particulate phases were constant, and (iii) PAH fingerprints exhibited a similar pattern for all catchments. Moreover, an evaluation of the contribution of DWF, runoff and erosion of sewer deposits to WWF load was established. According to the hypothesis on the runoff concentration, the contributions were evaluated at 14, 8 and 78%, respectively, at the scale of the Marais catchment. For all the catchments, the runoff contribution was found quite constant and evaluated at approximately 10%. The DWF contribution seems to increase with the catchment area, contrary to the sewer erosion contribution, which seems to decrease. However, this latter still remains an important source of pollution. These first trends should be confirmed and completed by more investigations of rain events. PMID- 17120650 TI - Polycyclic aromatic hydrocarbons and metals in snow along a highway. AB - Snow quality and its variations due to distance from the road were studied. Also, how the snow quality changes over time during the melting period was discussed. Snow samples were collected at three occasions during the winter of 2004. The samples were taken along a highway in the Lulea region, Sweden, with an average daily traffic load of 9200 vehicles. Snow samples were taken perpendicular to the road and at different distances. The snow samples were analysed for metals and polycyclic aromatic hydrocarbons (PAH). Also, weather parameters such as temperature, precipitation, and wind speed and wind direction were measured. The highest total metal and PAH concentrations were found at the sample site closest to the road and at the end of the season. Before the melting period started, 42 57% of the total amount of metals and PAH were found in the first 1 m of the snow pack. This information could be valuable when one is discussing how to achieve sustainable snow-handling management. PMID- 17120651 TI - Prolonged deposition of heavy metals in infiltration facilities and its possible threat to groundwater contamination. AB - A field investigation of infiltration facilities, built two decades ago in Tokyo, was carried out and sediment samples were collected from 12 infiltration inlets of three different locations. Heavy metals contents in the inlet sediment, road dusts and soils samples were analysed and compared. The particle size distribution analysis showed its variation in depth as well as from inlet to inlet. The nature of organic substances present in sediment found changes in particle sizes as well as in depth. The heavy metals content in the sediment samples ranged from 6-143 (Cr), 1-84 (Ni), 49-331 (Cu), 210-2186 (Zn) and 2-332 (Pb) microg/g. The heavy metal content ranges were similar to road dust, which indicated road dust as a possible source for sediment for the infiltration inlets. The lower heavy metals content in many sediment samples than the soil indicated possible release/desorption of heavy metals under newly created environments such as an anaerobic environment. Among the heavy metals there was a relatively good relationship between Cu and Zn, indicating the existence of their common sources. PMID- 17120652 TI - Integrated modelling of two xenobiotic organic compounds. AB - This paper presents a dynamic mathematical model that describes the fate and transport of two selected xenobiotic organic compounds (XOCs) in a simplified representation of an integrated urban wastewater system. A simulation study, where the xenobiotics bisphenol A and pyrene are used as reference compounds, is carried out. Sorption and specific biological degradation processes are integrated with standardised water process models to model the fate of both compounds. Simulated mass flows of the two compounds during one dry weather day and one wet weather day are compared for realistic influent flow rate and concentration profiles. The wet weather day induces resuspension of stored sediments, which increases the pollutant load on the downstream system. The potential of the model to elucidate important phenomena related to origin and fate of the model compounds is demonstrated. PMID- 17120653 TI - An elutriation apparatus for assessing settleability of combined sewer overflows (CSOs). AB - An elutriation apparatus was proposed for testing the settleability of combined sewer outflows (CSOs) and applied to 12 CSO samples. In this apparatus, solids settling is measured under dynamic conditions created by flow through a series of settling chambers of varying diameters and upward flow velocities. Such a procedure reproduces better turbulent settling in CSO tanks than the conventional settling columns, and facilitates testing coagulant additions under dynamic conditions. Among the limitations, one could name the relatively large size of the apparatus and samples (60 L), and inadequate handling of floatables. Settleability results obtained for the elutriation apparatus and a conventional settling column indicate large inter-event variation in CSO settleability. Under such circumstances, settling tanks need to be designed for "average" conditions and, within some limits, the differences in test results produced by various settleability testing apparatuses and procedures may be acceptable. Further development of the elutriation apparatus is under way, focusing on reducing flow velocities in the tubing connecting settling chambers and reducing the number of settling chambers employed. The first measure would reduce the risk of floc breakage in the connecting tubing and the second one would reduce the required sample size. PMID- 17120654 TI - Design of a retention tank: comparison of stormwater quality models with various levels of complexity. AB - Stormwater quality simulation models are useful tools for the design and management of sewer systems. Modelling results are highly sensitive to experimental data used for calibration. This sensitivity is examined for three modelling approaches of various complexities (site mean concentration approach, event mean concentration approach and build-up, washoff and transport modelling approach) applied to a typical case study (design of a dry detention tank), accounting for the variability of calibration data and their effect on simulation results. Calibrated models with different calibration data sets were used to simulate 3 years of rainfall with different retention tank specific volumes. Annual pollutant load interception efficiencies were determined. Simulations results revealed (i) that there is no advantage in using the EMC model compared to the SMC model and (ii) that the BWT model resulted in higher design ratios than those given by the SMC/hydraulic approach. For both EMC and BWT models, using an increasing number n of events for calibration leads to narrower confidence intervals for the design ratios. It is crucial for design ratios to account for successive storm events in chronological order and to account for the maximum allowable flow to be transferred to the downstream WWTP. PMID- 17120655 TI - Assessment of CSO loads--based on UVIVIS-spectroscopy by means of different regression methods. AB - The use of UV/VIS-spectroscopy for water quality measurements is based on the solution of the correlation between the surrogate parameter absorbance and the resulting equivalence parameters. The coherence of absorbance and equivalence parameters (CODtot, CODsol, TSS) is solved in this paper with different regression methods. The correlation of absorbance and concentrations are analysed based on linear regression methods, model tree regressions, multivariate regression methods and support vector machines using sequential minimal optimisation algorithm. For this purpose the regression methods are calibrated on three 24hours measurement campaigns of a combined sewer measurement station situated in the combined sewer overflow chamber in Graz (Austria). The online measurement station has been conveying data for more than 2 1/2 years up to now. Finally, the load calculation based on the different regression methods and its comparison demonstrate that an apparently complex model does not inevitably lead to accurate concentration values due to possible model overfitting. Hence, the paper points out the possibilities and the drawbacks of spectroscopy measuring in sewers and the arising concentration values. PMID- 17120656 TI - Assessment of uncertainties in the modelling of CSOs. AB - Engineers generally use simulation tools such as SWMM, MOUSE, HYDROWORKS, etc. to simulate the operation of sewer systems. Equations used by these models to represent the hydraulics of sewer systems are well known as well as their performance. Thus assessment of the performance needs to suppose that the user is an expert who is able to describe each facility in the most accurate way according to the model. In Western Europe, important parts of sewer systems are old and combined structures. Such structures very often include very complicated special structures, especially within CSO facilities. Experience shows that the performance limits of the models do not rely on the accuracy of the equations, but on the ability of the modeller to describe these special structures properly. The objective of this research is to assess the part of errors and uncertainties which originate from modellers' inconsistencies in the description of special structures. Even if the results are obtained with specific software (CANOE), results can easily be generalised to other models, since the equations used (Barre de Saint Venant, etc.) are almost the same. PMID- 17120657 TI - The role of in situ unit operation/process infiltration treatment on partitioning and speciation of rainfall-runoff. AB - Management decisions regarding the potential fate and toxicity of anthropogenic metal elements requires a knowledge of metal partitioning and speciation as mediated by in situ control systems (ICS). This study focussed on Cd, Zn, Cu and Pb, common anthropogenic metal elements generated in the built environment and examined the influence of variable event-based hydrology and passive ICS by an engineered partial exfiltration reactor (PER) system on partitioning and speciation. The feasibility and efficiency of the PER as an in situ stormwater runoff control strategy to attenuate levels of metal elements was evaluated. Results indicate that higher rainfall intensity resulted in higher dissolved fraction (fd) values for influent Zn, Cu and Cd, but did not have a significant influence on partitioning of Pb. Speciation indicated that divalent fractions of Cd, Zn, Cu and Pb were changed marginally by the PER. Cu and Pb mainly complexed with organic matter in the influent, while carbonate complexes of these metals in the effluent significantly increased. The PER consistently and statistically reduced all loadings of Cd, Cu, Pb and Zn for all examined events, whether on a mass or concentration basis. As an unsaturated flow unit operation/process the PER can efficiently remove ionic or complexed forms of metal elements. Such UOPs are a potential quality and quantity ICS strategy for rainfall-runoff. PMID- 17120658 TI - Filter media for stormwater treatment and recycling: the influence of hydraulic properties of flow on pollutant removal. AB - Improved urban water management in Australia is of national importance. Water resources are stretched and urban runoff is a recognized leading cause of degradation of urban waterways. Stormwater recycling is an option that can contribute to easing these problems. Biofilters are effective structural stormwater pollution control measures with the potential for integration into stormwater treatment and recycling systems. However, premature clogging of biofilters is a major problem, with resulting decreased infiltration capacity (and hence the volume of stormwater the system can detain) and increased detention time. This paper presents preliminary findings with respect to the effect of clogging on pollutant removal efficiency in conventional stormwater filter media. A one-dimensional laboratory rig was used to investigate the impact of clogging on pollutant removal efficiency in a conventional biofiltration filter media (gravel over sand). Both the individual gravel layer and the overall multi-filter were highly efficient at removing suspended solids and particulate associated pollutants. This removal efficiency was consistent, even as the filters became clogged. Removal of dissolved nutrients was more variable, with little reduction in concentrations overall. Although preliminary, these results challenge the concept that increased detention time improves the treatment performance of stormwater filtration systems. PMID- 17120659 TI - The development of geotextiles incorporating slow-release phosphate beads for the maintenance of oil degrading bacteria in permeable pavements. AB - The development of a self-fertilising geotextile mat designed to provide a sustained slow-release of required inorganic nutrients for the growth of oil degrading microorganisms in porous pavement systems (PPS) is reported. The system comprises a geotextile spun from polymer fibres containing spherical phosphated polymer beads that release phosphate upon contact with water at a desirable level for microbial growth. Initial results using model PPS have shown that the self fertilising geotextile system works extremely effectively as increased microbial activity has been observed throughout the experiment, illustrating that the oil degrading bacteria can effectively utilise this polymer composite as a suitable nutrient source. PMID- 17120660 TI - Infiltration of a copper roof runoff through artificial barriers. AB - On-site infiltration of a copper roof runoff may contribute to deterioration of the ground and ground water. To avoid such a negative effect the performance of two different technical systems, equipped with four different barrier materials, regarding copper elimination was examined in a field study. During the period March 2004 to January 2005, 16 rain events were examined. Copper concentrations between 200 and 11,000 microg/L in the roof runoff during a rain event were observed. The cover material of the roof and the drainage system were responsible for the high concentrations of copper in the roof runoff. It was evident that roof aspects facing towards the wind direction were receiving higher rainfall, thus were establishing higher copper runoff rates. The retention facilities have reached a performance of up to 97% regarding copper elimination. PMID- 17120661 TI - Development and investigation of a pollution control pit for treatment of stormwater from metal roofs and traffic areas. AB - Source control by on-site retention and infiltration of stormwater is a sustainable and proven alternative to classical drainage methods. Unfortunately, sedimentary particles and pollutants from drained surfaces cause clogging and endanger soil and groundwater during long-term operation of infiltration devices. German water authorities recommend the use of infiltration devices, such as swales or swale-trench-systems. Direct infiltration by underground facilities, such as pipes, trenches or sinks, without pretreatment of runoff is generally not permitted. Problems occur with runoff from metal roofs, traffic areas and industrial sites. However, due to site limitations, underground systems are often the only feasible option. To overcome this situation, a pollution control pit was developed with a hydrodynamic separator and a multistage filter made of coated porous concrete. The system treats runoff at source and protects soil, groundwater and receiving waterways. Typically, more than 90% of the pollutants such as sedimentary particles, hydrocarbons and heavy metals can be removed. Filters have been developed to treat even higher polluted stormwater loads from metal roofs and industrial sites. The treatment process is based on sedimentation, filtration, adsorption and chemical precipitation. Sediments are trapped in a special chamber within the pit and can be removed easily. Other pollutants are captured in the concrete filter upstream of the sediment separator chamber. Filters can be easily replaced. PMID- 17120662 TI - Pollutant removal efficiency of alternative filtration media in stormwater treatment. AB - Sorption experiments were used to assess the ability of various materials (sand, compost, packing wood, ash, zeolite, recycled glass and Enviro-media) to remove heavy metal contaminants typically found in stormwater. Compost was found to have the best physicochemical properties for sorption of metal ions (Cu, Zn and Pb) compared with sand, packing wood, ash, zeolite and Enviro-media. The compost sorption of these metal ions conformed to the linear form of the Langmuir adsorption equation with the Langmuir constants (q,) for Zn(ll) being 11.2 mg/g at pH 5. However, compost was also found to leach a high concentration of dissolved organic carbon (DOC, 4.31 mg/g), compared with the other tested materials. Various combinations of sand, compost and other materials were observed to have excellent heavy metal removal (75-96% of Zn and 90-93% of Cu), with minimal DOC leaching (0.0013-2.43 mg/g). The sorption efficiency of the different Enviro-media mixes showed that a combination of traditional (sand) and alternative materials can be used as an effective medium for the treatment of dissolved metal contaminants commonly found in stormwater. The application of using recycled organic materials and other waste materials (such as recycled glass) also provides added value to the products life cycle. PMID- 17120663 TI - Colloidal speciation of heavy metals in runoff and interstitial waters of a retention/infiltration pond. AB - The thickness of non-saturated zone and physico-chemical conditions are important parameters to assess the impact of infiltration ponds on water resources with respect to heavy metals transfer. As changes in physico-chemical parameters of solutions have a strong impact on the mobility of colloidal phases in sediments and soils, the colloidal facilitated transfer of heavy metals has to be investigated. Therefore, this study focuses on the characterization of runoff, surface and interstitial waters in a retention/infiltration pond collecting runoff waters of a bridge near Nantes. Physico-chemical parameters and chemical analyses were performed on the waters during about one year. The separation of dissolved and colloidal fractions was carried out by filtration and ultrafiltration for one sample of surface and interstitial waters. Until now, the runoff waters were only filtered at 0.45 microm. The comparison of physico chemical data shows that the minor variations of runoff water parameters are mitigated in basin and in soils but strong variations impact the composition of interstitial waters. High concentrations of zinc, copper and still of lead are measured in runoff. Lead and cadmium seem to be associated to colloidal and particulate fractions while zinc, copper, nickel and chromium are distributed in all fractions. PMID- 17120664 TI - Occurrence and relative abundance of mosquitoes in stormwater retention facilities in North Carolina, U.S.A. AB - Throughout the 2004 mosquito season, 52 stormwater retention facilities were sampled to characterize the seasonal occurrence and relative abundance of mosquito species in relation to the structural complexity and biological diversity of the facilities. The three different types of facilities included standard wet ponds (n=20), innovative ponds (n=14), and wetland ponds (n=18). All retention structures were sampled at the beginning, middle and end of the mosquito season so that seasonal changes in mosquito production could be characterized. Overall samplings, mosquitoes were collected from 34% of the retention structures. Fourteen species representing 7 genera were collected, but only 5 species were commonly collected in all three types of stormwater management facilities. In general, the seasonal prevalence and relative abundance of mosquito species did not vary among three types of retention structures. A significant association (P < 0.01) between the presence of mosquito larvae or pupae and the absence of mosquitofish was found for innovative and wetland stormwater retention facilities but not for standard retention facilities (P > 0.05). PMID- 17120665 TI - Transfer of hydrophobic contaminants in urban runoff particles to benthic organisms estimated by an in vitro bioaccessibility test. AB - An in vitro bioaccessibility test was applied for assessing the transfer of polycyclic aromatic hydrocarbons (PAHs) present in road dust, into benthic organisms living in a receiving water body. The road dust is supposed to be urban runoff particles under wet weather conditions. Sodium dodecyl sulfate (SDS) solution was used as a hypothetical gut fluid. Pyrene, fluoranthene and phenanthrene were the main PAH species in the SDS extractable fraction of road dust, as well as the whole extract. Benzo(ghi)perylene showed relatively low concentrations in the SDS extract in spite of a high concentration in the original dust. The PAH composition in benthic organisms (polychaetes) did not correspond with that of the surrounding sediment and the PAHs detected were also detected in high concentrations in the SDS extract of road dust. When testing the toxicity of the extracted contaminants by a standardised algal toxicity test, SDS extracts of a detention pond sediment showed higher toxicity than the pore water of the corresponding sediment. Sediment suspension showed a comparative toxicity with 0.1% SDS extract. From the results, the in vitro bioaccessibility test seems more suitable to evaluate the exposed contaminants than the traditional organic solvent extraction method and the SDS extracted fraction is applicable to toxicity tests reflecting the digestive process. PMID- 17120666 TI - Dynamics of the nitrogen transformation in a shallow stream and possible interventions. AB - This paper presents the assessment of the efficiency of the main biological nitrogen transformation processes in a shallow well-oxygenated river and conditions under which they are active and stabilise. The process dynamics was studied with the help of mathematical modelling of 2 years on-line data series measured in a reach of the Toess River, Switzerland. The algal nitrogen uptake was very stable and unaffected by most but frequent flood events. Daylight photosynthetic nitrogen uptake stabilised at 6 mgN mstreambed(-2) h(-1) (15 degrees C), dark uptake on storage products at rates of 0.5-2.5 mgN mstreambed( 2) h(-1). Nitrogen uptake by heterotrophic bacteria in the hyporheic zone was relatively constant at a level of 1.5-3.5 mgN mstreambed(-2) h(-1). Streambed nitrification could establish only during periods with average an daily concentration of at least 0.3 g(NH4-N) m(-3) in river water for several weeks. The maximum nitrification rate was 35 mgN mstreambed(-2) h(-1) for 3 g(NH4-N) m( 3). The effects of reduced nitrification in the WWTP and of river banks shading on a sudden ammonium peak were simulated. A river reach endangered by ammonium spills should be kept open to sun to favour ammonium uptake by algae. In-stream nitrification reduces ammonium peaks efficiently but leads to toxic nitrite concentrations. PMID- 17120667 TI - Distribution of heavy metals in freshwater ecosystem of a small stream impacted by urban drainage. AB - This paper focuses on the study of heavy metals' remobilisation possibilities and changes of distribution coefficients leading to remobilisation in a stream as a result of changing conditions caused by urban drainage. The paper also reports consequent changes of bioaccumulation coefficient for different species of the benthic community, which seems to be the best indicator of aquatic community quality in small urban streams. The paper presents results obtained during field monitoring as well as results obtained during laboratory experiment. The field monitoring identified in the study area (the Botic Creek) three heavy metals (Cu, Zn and Pb), which are the main sources of toxicological risk in this area. Based on the monitoring, these elements were used for laboratory experiment, where their behaviour during changing conditions was studied. The experiment shows that changing conditions in the stream, caused by urban drainage, significantly impact the fate of heavy metals in aquatic ecosystems and that these elements have different preferences to bind to a particular geochemical fraction of sediment, which consequently means different bioavailability. PMID- 17120668 TI - Matching the critical portion of the flow duration curve to minimise changes in modelled excess shear. AB - Hydrologic and hydraulic modeling in the USEPA Stormwater Management Model (SWMM) were used to examine the effectiveness of typical stormwater management practices in reducing the potential for stream erosion. Fifty-year continuous simulations were used to produce flow duration curves and stream erosion rates for a variety of critical shear stress values representative of both cohesive and non-cohesive sediments. An excess shear stress erosion potential index was used to evaluate changes in erosion between undeveloped conditions of a 10 hectare watershed and four variations of post-development stormwater control. Evaluation of flow duration curves showed that when development takes place, the duration of mid- to low-range discharges increase significantly, especially when detention practices are applied. In channels with low entrainment thresholds for bed and bank materials, e.g. sands and highly erodible clays, the significant increase of the duration of mid- to low-range discharges results in erosion potential index values greater than two regardless of the detention practices used. Overcontrol detention resulted in erosion potential index values of less than one, indicating a loss of erosion potential for bed materials such as most gravels (d(s) > 6 mm) and resistant clays that have critical shear stress values greater than four Pa. PMID- 17120669 TI - Water quality criteria for total suspended solids (TSS) in urban wet-weather discharges. AB - Total suspended solids (TSS) from combined sewer overflows (CSOs) and stormwater discharges represent a crucial parameter for evaluating wet-weather pollution in urban areas. In fact, the increase of TSS in water during rain events can have ecotoxic effects on aquatic organisms. Furthermore, major potentially harmful substances such as heavy metals, PAHs and organic matter are adsorbed onto TSS and later settle on sediment. Water quality criteria for TSS consequently enable the risk of wet-weather pollution to be assessed, for instance to avoid detrimental effects on aquatic organisms. The criteria proposed in this study cover the short-term impact of TSS on fish (acute quality criteria AQC), taking into account the duration of their exposure in the receiving water. The concentration-exposure duration-effect curve proposed here thus predicts "ill effects" on fish for different exposure times and TSS concentrations. The ecotoxic effects of adsorbed pollutants are also taken into account with an additional safety factor. We implement this TSS criteria in a software that allows us to estimate the number of rain events that exceed a given morbidity threshold for fishes per year. PMID- 17120670 TI - Estimation of solids loadings to rainfall-runoff unit operations using a unit pollutograph concept for source area watersheds. AB - Source area watershed in the built environment deliver significant loads of solids and constituents such as metals, organics and phosphorus associated with solids. The ability to model the mass delivery process of solids in small urban watersheds is essential for advancement of rainfall-runoff quantity and quality control design in the built environment. In this study the hydrologic concepts of the unit hydrograph (UH) and instantaneous unit hydrograph (IUH) were used to support the concepts of a unit pollutograph (UP) and an instantaneous unit pollutograph (IUP). These latter two concepts were developed as analogous concepts for estimation of solids mass transport loading as a pollutograph from source area watersheds. Relationships between solids mass and hydrologic volume were based on relationships that expressed either a first-order or zero-order relationship between solids mass and volumetric transport. The resulting concepts were applied to historical rainfall-runoff events where the hydrologic and water quality measurements had been conducted. From this development a site mean IUH and IUP determined from the rainfall-runoff record and water quality data, the corresponding site mean UH and UP were developed for solids. The hydrographs and solid pollutographs generated by convolution of the UH and UP compared well with measurements, indicating the feasibility of the concept. PMID- 17120671 TI - Assessment of WWTP design and upgrade options: balancing costs and risks of standards' exceedance. AB - Numerical models can be used to evaluate design and upgrade scenarios of urban wastewater systems on the basis of their ecological consequences. The objective of this paper is to illustrate a systematic procedure of system design/upgrade. This procedure consists of the following steps: (1) data collection and data reconstruction; (2) model building and calibration; (3) evaluation of scenarios; (4) uncertainty assessment. In contrast to conventional practice, this approach allows to choose the most appropriate trade-off between cost of measures and risk of non-compliance with regulatory limits. An example of its application dealing with the assessment of WWTP design and upgrade options is provided. Results show that by reducing the tank volumes compared to conventional design procedures, costs can be reduced sensibly while the risk of not meeting legislative requirements are only slightly increased. PMID- 17120672 TI - Integrated urban water modelling with uncertainty analysis. AB - In the last twenty years, the scientific world has paid particular care towards the problems that involve the environment. Accordingly, several researches were developed to describe phenomena that take place during both wet and dry periods and to increase the knowledge in this field. In particular, attention was addressed towards the problems linked with receiving water body pollution because of the impact of rain water in the urban environment. In order to obtain a good description of the problem, it is important to analyse both quantity and quality aspects connected with all the transformation phases that characterise the urban water cycle. Today, according to this point, integrated modelling approach is spreading, aiming to find solutions to improve the quality characteristics of the receiving water body. Because several models are connected together for analysing the fate of pollutants from the sources on the urban catchment to the final recipient, classical problems connected with the selection and calibration of parameters are amplified by the complexity of the modelling approach increasing the uncertainty and reducing the reliability connected with a model's application. For this reason, a parsimonious integrated modelling approach has been developed and its uncertainty has been evaluated adopting the well known GLUE framework. For the purpose of the study, the uncertainty analysis has been applied to a "semi-hypothetic" case study obtained connecting Fossolo catchment (Bologna-Italy) to the Oreto river near Palermo (Italy). PMID- 17120673 TI - Random number generator or sewer water quality model? AB - Integrated urban drainage modelling and environmental impact assessment require sewer emission models to be linked with submodels for treatment infrastructure and receiving rivers. The uncertainty in current water quality modelling is, however, huge, and environmental impact assessment looses more and more credibility. Based on an integrated modelling case for a combined sewer-WWTP river system, it is shown in the paper that the integrated model does not produce more accurate results in comparison with the random simulation of emission concentrations from a frequency distribution. This should, however, not pose a serious problem as in most applications of impact assessment, model results are not needed in real time but in statistical terms. Further investigation makes clear that detail/sophistication in water quality modelling is not so important, but that more focus has to be given to long-term simulations, the use of parsimonious models and model validation based on concentration frequencies. PMID- 17120674 TI - Integrating groundwater into urban water management. AB - The management of urban groundwater resources is directly linked to urban water supply and drainage concepts. A proper integration of groundwater into urban water management plans is recommended for long-term planning. The paper describes the development of a new modelling suite which addresses the urban water and solute balance in a holistic way. Special focus has been placed on the assessment of the impact of sewer leakage on groundwater in four case study cities. Tools for the prediction of sewer leakage including the assessment of uncertainties are now available. Field investigations in four European case study cities were able to trace the influence of sewer leakage on urban groundwater using microbiological indicators and pharmaceutical residues. PMID- 17120675 TI - The scope of integrated modelling: system boundaries, sub-systems, scales and disciplines. AB - Integrated modelling has become an urgent issue of urban drainage and wastewater treatment planning. The scope of integrated modelling, system boundaries and disciplines to be involved are addressed in view of future developments and new paradigms in urban drainage, demanding the inclusion of the full urban water cycle. A system analysis is demonstrated to identify relevant sub-systems and components, processes and interactions within the urban water system. The permissibility to exclude subsystems or neglect interactions is evaluated. Integrated modelling of urban water system is characterised as an ambitious task in regard to system complexity, heterogeneous scales and interface problems. The methodical status quo is characterised in preliminary approaches towards integrated modelling. It is concluded that it does not seem promising to create and apply one entity model for the scope of integrated urban water modelling. Instead, the development of adequate and efficient IT frameworks is identified as the key issue of integrated modelling. Harmonising interfaces to facilitate the linking of existing models is presented as the objective of a European research project HarmonlT and the U.S. EPA Multimedia Integrated Modelling System project MIMS. PMID- 17120676 TI - Implementation impediments to institutionalising the practice of sustainable urban water management. AB - It is now well accepted that there are significant challenges to realising the widespread and self-sustaining implementation of sustainable urban water management. It is argued that these challenges are entrenched within the broader socio-political framework, yet often unsuccessfully addressed within the more narrow scope of improving technical knowledge and design capacity. This hypothesis is investigated through a comparative analysis of three independent research projects investigating different dimensions of the water cycle, including stormwater management in Australia and sanitary waste management and implementation of innovative technologies in the U.K. The analysis reveals significant and common socio-political impediments to improved practice. It was evident that the administrative regime, including implementing professionals and institutions, appears to be largely driven by an implicit expectation that there is a technical solution to solve water management issues. This is in contrast to addressing the issues through broader strategies such as political leadership, institutional reform and social change. It is recognised that this technocratic culture is inadvertently underpinned by the need to demonstrate implementation success within short-term political cycles that conflict with both urban renewal and ecological cycles. Addressing this dilemma demands dedicated socio-technical research programs to enable the much needed shift towards a more sustainable regime. PMID- 17120677 TI - Centralised urban wastewater reuse: what is the public attitude? AB - Public support is crucial for successful implementation of wastewater reuse projects. This paper analyses the findings of a questionnaire-type survey (256 participants) conducted to determine the attitude of Israeli urban public towards possible urban reuse options. The paper summarises the support/objection to 13 reuse options and the correlation between support and environmental awareness and perceptions. Results show that a high proportion of the participants supported options perceived as low-contact, such as irrigation of public parks (96%), sidewalk landscaping (95%) and use in the construction industry (94%), while higher-contact reuse options found less support (e.g. commercial launderettes, 60%). No correlation was found between biographical characteristics and support (education, gender, income, age). Based on the results, public campaigns in Israel should focus on disseminating information regarding wastewater treatment technologies, discuss health related issues, highlight the positive economic impact of water reuse and generate a positive public opinion, as these factors tend to influence individuals to support reuse projects. PMID- 17120678 TI - The perception of flood risk and water nuisance. AB - In this paper we applied the psychometric paradigm to validate a questionnaire that assesses the risk perception characteristics of flooding and water nuisance. The state-trait anxiety inventory was used as a bench mark to determine whether perceptions are related to anxiety characteristics. A focus group was used to further validate the questionnaire. Factor analyses of 49 questionnaires identified eight flooding factors (explained variance 74%) and three water nuisance factors (explained variance 62%). Internal consistencies of the obtained scales were moderate to high. Like in the perception of external safety risks, "dread" seems to be the most important concept binding different characteristics. Although dread towards both flooding and water nuisance is rather low, it seems more present in the latter case. Furthermore, the extent of dread for water nuisance seems related to someone's state anxiety. In both cases awareness of "increasing risks" is clearly present, and we find the characteristics "(no) dread", "(un)controllable situation" and "does not affect me" to be related. PMID- 17120679 TI - Indicators of hazard, vulnerability and risk in urban drainage. AB - An alternative definition of risk is proposed as risk being a function of the hazard, which is related to the risk source and the vulnerability, which is related to the risk object. The same hazard will not cause the same effect on all risk objects. Therefore, vulnerability is introduced as a system-dependent property to be the link between the hazard and the effect so that the combination of the occurrence of a hazard and the vulnerability of an object results in the effect. In risk communication indicators are helpful since they help to simplify the message that has to be communicated. Three examples (pluvial flooding of sewers, dissolved oxygen depletion in streams and discharge of chemicals to receiving waters) show that dependent on the risk problem possibilities for risk reduction lies either at the risk source or at the risk object. Therefore, it is important to have indicators that can be used when the possibilities of risk reduction are analysed. PMID- 17120680 TI - A decision support framework for sustainable urban water planning and management in new urban areas. AB - The paper discusses issues of decision support within the context of sustainable development and more specifically sustainable water cycle management to provide a context and a rationale for the decision support approach adopted within an on going U.K. EPSRC-funded project, WaND. The paper proposes a set-up for a flexible, upgradeable, efficient and modular decision support framework and associated tools. Furthermore, the paper presents early prototypes of three decision support tools developed within the proposed framework including initial results for one of them. PMID- 17120681 TI - "Triple-bottom-line" assessment of urban stormwater projects. AB - New guidelines have been developed and trialled in Australia to assist urban stormwater managers to assess options for projects that aim to improve urban waterway health. These guidelines help users to examine the financial, ecological and social dimensions of projects (i.e., the so-called "triple-bottom-line"). Features of the assessment process described in the guidelines include use of multi criteria analysis, input from technical experts as well as non-technical stakeholders, and provision of three alternative levels of assessment to suit stormwater managers with differing needs and resources. This paper firstly provides a background to the new guidelines and triple-bottom-line assessment. The assessment methodology promoted in the new guidelines is then briefly summarised. This methodology is compared and contrasted with European guidelines from the "SWARD" project that have been primarily developed for assessing the relative sustainability of options involving urban water supply and sewerage assets. Finally, the paper discusses how assessment methodologies that evaluate the financial, ecological and social dimensions of projects can, under some circumstances, be used to evaluate the relative progress of options for urban water management on a journey towards the widely pursued, but vaguely defined goal of "sustainable development". PMID- 17120682 TI - Dynamic least-cost optimisation of wastewater system remedial works requirements. AB - In recent years, there has been increasing concern for wastewater system failure and identification of optimal set of remedial works requirements. So far, several methodologies have been developed and applied in asset management activities by various water companies worldwide, but often with limited success. In order to fill the gap, there are several research projects that have been undertaken in exploring various algorithms to optimise remedial works requirements, but mostly for drinking water supply systems, and very limited work has been carried out for the wastewater assets. Some of the major deficiencies of commonly used methods can be found in either one or more of the following aspects: inadequate representation of systems complexity, incorporation of a dynamic model into the decision-making loop, the choice of an appropriate optimisation technique and experience in applying that technique. This paper is oriented towards resolving these issues and discusses a new approach for the optimisation of wastewater systems remedial works requirements. It is proposed that the optimal problem search is performed by a global optimisation tool (with various random search algorithms) and the system performance is simulated by the hydrodynamic pipe network model. The work on assembling all required elements and the development of an appropriate interface protocols between the two tools, aimed to decode the potential remedial solutions into the pipe network model and to calculate the corresponding scenario costs, is currently underway. PMID- 17120683 TI - Life cycle optimisation for highway best management practices. AB - Highway runoff can cause a number of water quantity and quality problems. Stormwater management systems for highways have been developed based on a fast drainage for large storm situations. Non-point source pollution from highway runoff is a growing water quality concern. Stormwater quality control needs to be integrated into highway drainage design and operation to reduce the stormwater impacts on the receiving water. A continuous simulation/optimisation model for analysing integrated highway best management practices (BMPs) is presented. This model can evaluate the life cycle performance of infiltration and/or storage oriented highway BMPs. It can be directly integrated with spreadsheet optimisation tools to find the least cost options for implementing BMPs throughout a specified life cycle. PMID- 17120684 TI - Hydrologic regionalisation impacts on wet-weather control selection. AB - Continuous simulation is performed using the US Environmental Protection Agency (USEPA) Storm Water Management Model (SWMM) to evaluate regional differences around the United States in hydrologic and water quality performance of wet weather controls. Controls are characterised as being limited by peak inflow rate (i.e. any device with little or no storage, such as screens, filters and some proprietary devices) or by storage capacity (e.g., ponds, tanks). For flow limited devices, results are presented in the form of percentage of annual runoff volume captured (passing through the device) for a given inflow capacity. For storage-limited devices, results are presented in two forms: percentage of annual runoff volume captured as a function of unit basin size and drawdown (drain) time, and as a percentage of total suspended solids captured, for the same two variables. Regional differences are apparent, driven mainly by variations in rainfall patterns around the country. PMID- 17120685 TI - Comparison between two methodologies for urban drainage decision aid. AB - The objective of the present work is to compare two methodologies based on multicriteria analysis for the evaluation of stormwater systems. The first methodology was developed in Brazil and is based on performance-cost analysis, the second one is ELECTRE III. Both methodologies were applied to a case study. Sensitivity and robustness analyses were then carried out. These analyses demonstrate that both methodologies have equivalent results, and present low sensitivity and high robustness. These results prove that the Brazilian methodology is consistent and can be used safely in order to select a good solution or a small set of good solutions that could be compared with more detailed methods afterwards. PMID- 17120686 TI - [The role of biological agents in the treatment of psoriatic arthritis, literature review]. AB - INTRODUCTION: The recognition of the key pathogenetic role of TNF-alpha in psoriatic arthritis has made it possible to introduce new drugs in the treatment. TNF-alpha inhibitors available in Hungary (infliximab, etanercept, adalimumab) are potential therapies for patients who have not adequately responded to traditional disease-modifying antirheumatic drugs. OBJECTIVE: The aim of the study was to present the epidemiology and progression of psoriatic arthritis in Hungary based on national and international data, to assess the target population for biological therapy and to analyze their effectiveness, reviewing the available literature of randomized controlled trials. METHODS: The prevalence of psoriatic arthritis in Hungary was estimated using international data. Characteristics of psoriatic arthritis population were studied using the database of a rheumatology ward. A systematic literature search was performed to identify each relevant trial. A synthesis and comparison of the results from the 5 identified trials was performed and the average effect of biological agents was calculated. Both the fixed and the random effect model were used for the data synthesis; the results were probed with Mantel-Haenzel test. RESULTS: The prevalence of psoriatic arthritis is about 10.000-20.000 in Hungary. Average disease-duration was 10 years in the sample (n = 189), the most frequent was polyarticular form (51%). Regarding functional status the mean HAQ score was 1.46, with an average progression of 0.05 points/year. The trial data confirmed that biological agents are superior to placebo in improving symptoms (achieving ACR20); risk difference between biological treatments and placebo is 47% (RD = 0.47, 95% CI: 0.42-0.53). The biological treatment of 2 patients improves the status of 1 patient (NNT = 2.1 95% CI 1.9-2.4). There is no significant difference in efficacy between the three biological agents. CONCLUSIONS: TNF alpha inhibitors are effective treatments of psoriatic arthritis and are safe under strict medical control. The principles of indications, contraindications, administration and control have been worked out by the Rheumatology and Physical Medicine Board. PMID- 17120687 TI - [Vascular pleiosynergism--a new concept?]. AB - The coexistence of cardiovascular risk factors augments exponentially the relative risk value. Similarly, the parallel treatment of these risks proves multiplicative effectiveness. However, members of given drug classes show different capacity due to their variant main and other effects. Nowadays, among favorable "side" effects the pleoitropic form is in the focus of interest. In drugs with antiatherosclerotic outline the lipid lowering statins and the antihypertensive third generation calcium channel blocker amlodipin posses the widest pleiotropic profile. According to the newest molecular biological results both of them are able to enhance endothelial nitrogen-monoxid (NO) synthesis, to inhibit production of reactive oxide radicals and to reduce the migration and proliferation of smooth muscle cells, which mechanisms are independent from their main effects. Statins and amlodipin can increase the activity of eNOS by influencing the caveolin/eNOS complex. The presence of common targets in direct and indirect effects seems to be important in gaining greater efficacy. The valuable indirect therapeutical "cross reaction" can be delineated as pleiosynergism. The recent revisited results of ASCOT-LLA trial support in vitro observations. The difference in primary end points of atorvastatin/amlodipin versus atorvastatin/atenolol regimen against placebo was more than three times (3.3 x) greater in favour to amlodipin combination. The same benefits were seen in secondary end points. Because the capacity of the two schedules lowering blood pressure was very similar, the advantage can be based dominantly on the pleiosynergistic interaction between atorvastatin and amlodipin. PMID- 17120688 TI - [Inflammatory bowel disease and colorectal cancer]. AB - The colorectal cancer is one of the most serious complication of inflammatory bowel disease. Longer duration of the disease, extensive colitis, primary sclerosing cholangitis, family history of colorectal cancer are the main risk factors. The relative risk of colorectal cancer in ulcerative colitis is increased, however, there're marked geographically differences. Relative risk of colorectal cancer is 2.5 and small bowel cancer risk is 31.2 in Crohn's disease. There aren't prospective, randomized, controlled trials that definitively prove the benefit of surveillance of colorectal cancer in IBD. Colonoscopy improve the 5-year survival rate, however, there is no evidence for the reduction of mortality. Meta-analysis showed efficacy of mesalamine in the reduction of risk of colorectal cancer, but prospective trials have been missed. Chemoprotective role of other immunomodulators has not been proven yet. PMID- 17120689 TI - [Eczematous skin diseases]. AB - The skin, as one of the most important barriers of the human body, protects the inner homeostasis from the harmful environmental influences as well as physical, chemical and biological factors. When the impact of these factors exceeds the tolerance and reproducing capacity of the skin, pathological alterations will develop. If follows from this that dermatology can surely be considered to be a part of environmental medicine. Eczematous diseases are mostly pathological pictures of varied mechanisms developing as a result of environmental influences (irritants, contact allergens, microbes). Since their clinical appearance is similar, it is a serious professional challenge to diagnose them. In this article we present the clinical features, provoking factors of these skin diseases as well as therapeutical possibilities. PMID- 17120690 TI - [Do we use digitalis properly in the management of elderly patients suffering from the signs of chronic heart failure?]. AB - INTRODUCTION: The indications of digoxin therapy has been significantly narrowed and also the effective target therapeutic blood level has been decreased (0.9 micromol/L) compared to the previously desired one. OBJECTIVE: In this retrospective trial the data of 60 consecutive patients over 65 years (25 male, 35 female, mean age 77.3 +/- 5.0 y), hospitalized between 01. 01. 2002 and 31. 12. 2003 with a diagnosis of chronic heart failure and elevated (> 1.2 microg/I) serum level of digoxin, were analyzed. METHODS: Beside the analysis of the age, sex, serum level of digoxin and potassium, creatinine clearance value, symptoms and ECG-signs of digitalis intoxication, presence of atrial fibrillation, concomitant diseases and left ventricular ejection fraction value, the reasonability of digitalis treatment and therapy applied at the time of discharge (considering actual treatment guidelines) were also reviewed. RESULTS: At the admission mean serum level of digoxin was 2.1 +/- 0.9 microg/l. 20 patient's value (33.3%) was found above 2.2 microg/l. Symptoms characteristic for digitalis intoxication were observed in 28 patients. On the ECG performed at admission signs of digitalis effect/overdose were observed in 54 cases ("bigemin" ventricular extrasystoles, bradycardia, characteristic down-sloping ST depressions). The mean left ventricular ejection fraction of the patients (51.5 +/- 12.7%) did not suggest to a significant left ventricular systolic dysfunction. For the elevated serum level of digoxin the impaired renal function (mean creatinine clearance 42.9 +/- 21.3 mL/min) was responsible in most cases. In patients with the highest serum level of digoxin (n = 20, 3.2 +/- 0.7 microg/L) the creatinine clearance was even lower, 30.4 +/- 13.7 mL/min. During hospital treatment the administration of digitalis was found to be unnecessary and thus terminated in 44 patients. At the discharge only 16 patients were receiving digitalis, 14 of them digoxin and 2 patients digitoxin. CONCLUSIONS: The authors emphasize, that in case of elderly patients the indication and control of digitalis therapy requires greater precaution and tight doctor-patient cooperation. PMID- 17120691 TI - [What does fetal renal echogenicity mean in intrauterine growth retardation?]. AB - AIMS: The object of this study was to investigate the fetal renal and middle cerebral arterial blood flows in patients with normal and hyperechogenic kidneys during the fetal period. MATERIAL AND METHODS: Study group consisted of 82 pregnancies with intrauterine growth retardation. Group included pregnant women from the third trimester. Hyperechogenic medullae were detected in 17 out of 82 pregnancies with intrauterine growth retardation. RESULTS: Fetal renal hyperechogenicity appears to be an indicator of fetal arterial circulatory depression, correlated with pathological changes in the pulsatility index for the fetal renal arteries. The fetal renal arterial blood flow pulsatility index was significantly lower in hyperechogenic cases, while in the middle cerebral artery flow was in the normal range. This may also be an indication of subsequent intrauterine and neonatal complications, such as cesarean section because of fetal distress (47%), treatment in a neonatal intensive care unit (71%) or increased perinatal mortality (12%) in our cases. CONCLUSIONS: Detailed ultrasound of renal parenchyma and Doppler examination of renal and middle cerebral arteries appear to be an useful method in the prenatal diagnosis of reduced renal perfusion and of intrauterine hypoxia so as to detect possible pathological fetal conditions in utero. PMID- 17120692 TI - [Identification of a Meckel's diverticulum bleeding by urgent capsule endoscopy]. AB - A 10 year-old patient was admitted to our hospital due to severe gastrointestinal bleeding. His symptoms included hematochezia and fainting. Neither emergency upper endoscopy nor colonoscopy had identified the site of bleeding. During the colonoscopy they noticed that fresh blood entered the cecum from the ileum. An urgent capsule endoscopy was performed 8 hours after the patient admission. They placed the capsule with specific endoscopic technique into the proximal duodenum under general anesthesia. During 3,5 hours by the small bowel passage the capsule moved to the ileum and a focal bleeding lesion was detected at this site, suggesting the typical picture of bleeding Meckel's diverticulum. The patient was transfused by 1 unit of blood, with a minimum haemoglobin level 95 g/l. By surgery the ulcerated Meckel's diverticulum was identified and resected. Histopathology showed ectopic gastric mucosa. Surgical resection of the diverticulum resulted complete healing of this patient. The capsule endoscopy diagnosis was prompt, precise, and the all examinations were carried out within 24 hours. Involving urgent small bowel capsule endoscopy into the diagnostic workup of the obscure bleeding patient could considerably shorten the time to achieve a correct diagnosis and allowed the early institution of definitive treatment. By this one could spare a great number of costly alternative investigations with low diagnostic yield. PMID- 17120693 TI - The EXTRA difference: leadership development. PMID- 17120694 TI - The effect of content and physical properties of affective pictures on emotional responses. AB - The aim of this research was to study the influence of both the emotional content and the physical characteristics of affective stimuli on the psychophysiological, behavioral and cognitive indexes of the emotional response. We selected 54 pictures from the IAPS, depicting unpleasant, neutral, and pleasant contents, and used two picture sizes as experimental conditions (120 x 90 cm and 52 x 42 cm). Sixty-one subjects were randomly assigned to each experimental condition. We recorded the startle blink reflex, skin conductance response, heart rate, free viewing time, and picture valence and arousal ratings. In line with previous research (e.g., Bradley, Codispoti, Cuthbert, and Lang, 2001), our data showed an effect of the affective content on all the measurements recorded. Importantly, effects of the size of the affective pictures on emotional responses were not found, indicating that the emotional content is more important than the formal properties of the stimuli in evoking the emotional response. PMID- 17120695 TI - Cardiovascular reactivity in hypertensives: differential effect of expressing and inhibiting emotions during moments of interpersonal stress. AB - This study investigated cardiovascular reactivity of hypertensive adults during periods of emotional stress. Two types of instructions were given at different moments, to the same subject, either to express or to suppress feelings during role-play. Expressing, but not inhibiting, emotions elicited significantly higher reactivity during responding to negative scenes, followed by responding during the positive interactions. Blood pressure increases in both expressing and inhibiting conditions, were also found during the instruction periods. Results indicated that socially demanding situations represent a stressor whose effects may vary depending on whether or not respondents regulate expression of emotions. It is suggested that the difficulty in expressing emotions found in some hypertensive individuals may have the function of controlling or reducing blood pressure reactivity. PMID- 17120696 TI - Is sexual attraction independent of the instrumental and expressive traits? AB - Sexual attraction is an essential part of sex, just as the instrumental and expressive traits are the mainstay of gender. Various hypotheses concerning the dimensionality and independence versus dependence/overlapping of these core entities were tested. A group of 423 university students completed the Sexual Attraction Questionnaire (SAQ; Fernandez, Quiroga, and Rodriguez, 2006) and the 12-item Bem Sex Role Inventory (BSRI; Bem, 1974). Internal consistency and factor analyses (PAF) were conducted. The results support the dimensionality established for the SAQ and reveal some psychometric and conceptual weaknesses of the 12-item BSRI. The results also support the independence of the two cores: sexual attraction and the instrumental and expressive traits. The logical implications for the different viewpoints of the relations between sex and gender are discussed. PMID- 17120697 TI - Fathers and gender traditionalism: perception of inequality and life roles. AB - Men's discourse about the paternal role is changing significantly. Despite the fact that men still perceive themselves as being responsible for the family's economical protection and the children's discipline, they face increasing demands for more involvement in childcare. From this perspective, this work analyzes the traditional view of gender roles and the perception of inequality in a sample of 95 employed fathers, as well as the various levels of satisfaction with other life roles and their relevance as a function of some gender and sociodemographic variables. The results show that men do not maintain a very traditional gender ideology with regard to role distribution and they still consider the paternal role and feelings as the most important thing in their lives. However, the couple relationship gives them the most satisfaction. Level of traditionalism and age were the two significant predicting variables of perception of inequality of men and women. PMID- 17120698 TI - Psychopathological symptoms, social skills, and personality traits: a study with adolescents. AB - The purpose of this study is two-fold: (a) to study the concomitant relationships between psychopathological symptoms, cooperation, social skills, and other personality traits; and (b) to identify the predictive variables of psychopathological symptoms. The sample consists of 322 adolescents aged 14 to 17 years old. This study uses correlational methodology. In order to assess psychopathological symptoms, cooperation, social skills, and personality traits, the following scales are used: the Symptom Checklist (SCL-90-R; Derogatis, 1983), the Cooperativeness Scale (CS; Rigby, Cox, and Black, 1997), the MESSY social skills scale (Matson, Rotatori, and Helsel, 1983), and the TPT Personality Test (Corral, Pamos, Perena, and& Seisdedos, 2002). Pearson coefficients suggest that adolescents with many psychopathological symptoms have low levels of cooperative behaviors and social skills. They also score high in inappropriate assertiveness, impulsiveness, overconfidence, and jealousy-withdrawal and have low levels of emotional stability, sociability, and responsibility. Through multiple regression analyses, the following variables were identified as predictors of psychopathological symptoms: jealousy-withdrawal, low social integration, impulsiveness, and low self-concept. The role played by intervention programs promoting socio-emotional development to prevent psychopathological symptoms and enhance mental health is discussed. PMID- 17120699 TI - Psychophysiological training of operators in adaptive biofeedback cardiorhythm control. AB - A new individual computerized technique for psychophysiological training of operators before performing psychomotor activity on a computer model (psychomotor concentration and spatial orientation test) was developed. Qualitative criteria for the prediction of safe operator activity were formulated. Preliminary testing of operators' activity quality showed great dispersion of individual results: The amount of errors ranged from 0 to 56 and the rate of information processing varied from 1.01 up 3.56 bit/s. Subjects with initially identified respiratory sinus arrhythmia or synchronization caused by respiratory movements committed minimal recognition errors in initial stages at a high rate of information processing. The number of errors remained unchanged after the biofeedback cardio training cycle, with the rate of information processing increasing noticeably. Subjects without inherent harmonics developed harmonics after sessions of cardiorhythm biofeedback control, and their operator activity quality improved significantly, making fewer mistakes and increasing the rate of information processing. Biofeedback control led not only to the restoration of respiratory sinus arrhythmia, a favorable diagnostic sign, as revealed by cardiorhythmograms, but also resulted in improvement of the quality of operator activity. PMID- 17120700 TI - Conscious and unconscious sensory inflows allow effective control of the functions of the human brain and heart at the initial ageing stage. AB - The authors of the present article based their assumption on the concept that the sensory systems are the "windows to the brain" through which various functions of the human organism can be controlled. Comprehension of the fundamental mechanisms of the optimization of the sensory systems, brain, and cardiac functions has increased based on the prolonged sensory flows using conscious and unconscious aromatherapy and multimodal sensory activation. Sensory flow evoked stable systemic responses, including adaptive alteration of psycho-emotional state, attention, memory, sensorimotor reactions, intersensory interaction, visual information processing, statokinetic stability, and autonomic heart rhythm control. The efficacy and expediency of the use of sensory flow for non-medicinal correction of vital functions of the human organism at the initial stages of ageing was revealed. PMID- 17120701 TI - An immunochemical approach to model and long-term suppression of depressive and anxiety behavior. AB - The inhibitor of monoaminooxydase isatin and the ligand of B-receptors cholecystokinin-4 play a significant role in the suppression and induction of depressive and anxiety states. We induced the formation of auto-antibodies to these compounds against their conjugates with antigen-carrier by immunization of white rats. The result was long-term (more than 2 months) stimulation of depressive and anxiety behavior after immunization to isatin and, in contrast, the suppression of such behavior after immunization to cholecystokinin. The perspective of immunochemical approach to long-term correction of behavior is discussed. PMID- 17120702 TI - Factor structure of items from the personality Adjective Check List in a sample of normal adolescents. AB - We present the results of a principal-components analysis of the Personality Adjective Check List (PACL) at the item level, rather than at the scale level. The PACL was constructed by Strack (1987) to evaluate the personality scales proposed by Millon (1981). Data were obtained from a sample of normal adolescents. A scree test was applied to determine the number of factors to be extracted, and the retained factors were then rotated using a varimax procedure. Using the data from this second analysis, congruence coefficients were calculated by correlating the factor weights from the two analyses of men and women. Results reveal the existence in the whole sample of five unipolar personality factors, which we have called: Aggressive, Conscientious, Inhibited, Impulsive and Gregarious. These are not similar to those obtained from a sample of normal adults. PMID- 17120703 TI - Narrow-band 1, 2, 3, 4, 8, 16, 24, 32, 48, 64, and 96 cycles/360 degrees angular frequency filters. AB - We measured human frequency response functions for eleven angular frequency filters using a forced-choice procedure in a supra-threshold summation paradigm. Each of the eleven functions of 17 experimental conditions was measured 4-9 times among 12 observers. Results show that, for the arbitrarily selected filter phases, maximum summation effect occurred at test frequency for all filters. These results lead to the conclusion that there are narrow-band angular frequency filters operating in human visual system mostly through summation surrounded by inhibition at the specific test frequency ranges. Our previous suggestion (Simas and Santos, 2002), arguing that summation for the higher angular frequency filters should occur if background angular frequency contrast were set to a maximum of 5 times the test frequency threshold, was supported. PMID- 17120704 TI - The effect of white-noise mask level on sinewave contrast detection thresholds and the critical-band-masking model. AB - It is known that visual noise added to sinusoidal gratings changes the typical U shaped threshold curve which becomes flat in log-log scale for frequencies below 10c/deg when gratings are masked with white noise of high power spectral density level. These results have been explained using the critical-band-masking (CBM) model by supposing a visual filter-bank of constant relative bandwidth. However, some psychophysical and biological data support the idea of variable octave bandwidth. The CBM model has been used here to explain the progressive change of threshold curves with the noise mask level and to estimate the bandwidth of visual filters. Bayesian staircases were used in a 2IFC paradigm to measure contrast thresholds of horizontal sinusoidal gratings (0.25-8 c/deg) within a fixed Gaussian window and masked with one-dimensional, static, broadband white noise with each of five power density levels. Raw data showed that the contrast threshold curve progressively shifts upward and flattens out as the mask noise level increases. Theoretical thresholds from the CBM model were fitted simultaneously to the data at all five noise levels using visual filters with log Gaussian gain functions. If we assume a fixed-channel detection model, the best fit was obtained when the octave bandwidth of visual filters decreases as a function of peak spatial frequency. PMID- 17120705 TI - Geometrical structures of photographic and stereoscopic spaces. AB - Two experiments were conducted to investigate the geometrical structures of photographic and stereoscopic spaces. In Experiment 1, it was investigated how accurately photographic space reproduces real physical space, and the geometrical structure of photographic space was compared with that of visual space. As a result, the mapping function of distance between photographic and physical spaces (delta = ad(b)) shows that a and b range from 0.96-1.1 and 0.69-0.78. The mapping function of angle between photographic and physical spaces (phi = g phi(h)) shows that g and h range from 2.37-5.29 and 0.74-0.97. Further, photographic space has larger anisotropic property than visual space and photographic space may be hyperbolic. In Experiment 2, the geometrical structure of stereoscopic space was compared with that of visual space. It was found that stereoscopic space was almost the same as visual space. PMID- 17120706 TI - One-dimensional and multi-dimensional studies of the exocentric distance estimates in frontoparallel plane, virtual space, and outdoor open field. AB - The aim of this study is twofold: on the one hand, to determine how visual space, as assessed by exocentric distance estimates, is related to physical space. On the other hand, to determine the structure of visual space as assessed by exocentric distance estimates. Visual space was measured in three environments: (a) points located in a 2-D frontoparallel plane, covering a range of distances of 20 cm; (b) stakes placed in a 3-D virtual space (range = 330 mm); and (c) stakes in a 3-D outdoors open field (range = 45 m). Observers made matching judgments of distances between all possible pairs of stimuli, obtained from 16 stimuli (in a regular squared 4 x 4 matrix). Two parameters from Stevens' power law informed us about the distortion of visual space: its exponent and its coefficient of determination (R2). The results showed a ranking of the magnitude of the distortions found in each experimental environment, and also provided information about the efficacy of available visual cues of spatial layout. Furthermore, our data are in agreement with previous findings showing systematic perceptual errors, such as the further the stimuli, the larger the distortion of the area subtended by perceived distances between stimuli. Additionally, we measured the magnitude of distortion of visual space relative to physical space by a parameter of multidimensional scaling analyses, the RMSE. From these results, the magnitude of such distortions can be ranked, and the utility or efficacy of the available visual cues informing about the space layout can also be inferred. PMID- 17120707 TI - Distance perception in a natural outdoor setting: is there a developmental trend to overconstancy? AB - The main purpose of the present study was to investigate whether in natural environment, using very large physical distances, there is a trend to overconstancy for distance estimates during development. One hundred and twenty nine children aged 5 to 13 years old and twenty-one adults (in a control group), participated as observers. The observer's task was to bisect egocentric distances, ranging from 1.0 to 296.0 m, presented in a large open field. The analyses focused on two parameters, constant errors and variable errors, such as measuring accuracy and precision, respectively. A third analysis focused on the developmental pattern of shifts in constancy as a function of age and range of distances. Constant error analysis showed that there are two relevant parameters for accuracy, age, and range of distances. For short distances, there are three developmental stages: 5-7 years, when children have unstable responses, 7-11, underconstancy, and 13 to adulthood, when accuracy is reached. For large distances, there is a two-stage development: 5-11 years, with severe underconstancy, and beyond this age, with mild underconstancy. Variable errors analyses indicate that precision is noted for 7 year-old children, independently of the range of distances. The constancy analyses indicated that there is a shift from constancy (or slightly overconstancy) to underconstancy as a function of physical distance for all age groups. The age difference is noted in the magnitude of underconstancy that occurs in larger distances, where adults presented lower levels of underconstancy than children. The present data were interpreted as due to a developmental change in cognitive processing rather than to changes in visual space perception. PMID- 17120708 TI - An empirical test of formal equivalence between Emmert's law and the size distance invariance hypothesis. AB - Emmert's law and the size-distance invariance hypothesis have been said to be formally equivalent, provided that Emmert's law means that the perceived size of an afterimage is proportional to the perceived distance of the projected surface of the afterimage. However, there have been very few studies that have attempted to verify this formal equivalence empirically. We measured both the perceived size and distance of afterimages and real objects with the same proximal size. Nineteen participants projected afterimages of 1 deg in visual angle on the wall located at distances of 1 to 23 meters from the participants. They also observed real objects, disc-shaped and made from a sheet of Styrofoam board, with the same proximal size as that of the afterimages, which were located at the same physical distances as those of the wall on which the afterimages were projected. Each participant reproduced the apparent sizes of the afterimages and real objects using the reproduction method and estimated the apparent distances using the magnitude estimation method. When the mean apparent sizes of the afterimages and real objects, represented as a function of apparent distance, were fitted to a linear function, the slopes for the afterimages and real objects did not differ significantly. These results are interpreted as evidence for the formal equivalence of Emmert's law and the size-distance invariance hypothesis. PMID- 17120709 TI - Lightness compression and hue changes. AB - Two experiments were performed to relate the Bezold-Brucke (B-B) and lightness compression effects. The first used a calibrated screen to present an achromatic luminance staircase. In addition, it reproduced, the methodology and the essential aspects the lightness compression effect discovered by Cataliotti and Gilchrist (1995). That is, observers perceived a truncated grey scale (from white to medium grey) when the staircase was the only stimulation in the near background (Gelb condition), but not when presented on a Mondrian background, because of the high articulation level provided by this background. Experiment 1 design also included two other backgrounds that produced a partial compression effect. In Experiment 2, two chromatic staircases were used. Employing a naming task, changes in hue perception were only observed for the susceptible staircase. The observed changes were of two types. First, for the full staircase presentations, a Gelb background produced maximum lightness compression (more similarity in the lightness of the staircase stimuli) and, also, a minimum B-B effect (fewer differences in hue). Second, only for the Gelb condition, there were changes in the hue of the lowest luminance staircase stimuli depending on the staircase extension. Results are discussed in the framework of the anchoring theory of lightness perception. PMID- 17120710 TI - Influence of the instructions on the performance and establishment of memorization strategies in space judgments. AB - Studies of visual space perception have been assuming that people have an internal representation of the physical space that surrounds them. A variety of psychophysical procedures has been used in an attempt to measure the properties of visual space. The goal of the present study was to evaluate the accuracy of the mental representation and the strategies adopted to acquire and retain visuo spatial information of a configuration as a function of two types of instructions. Thirty-eight undergraduate and graduate students participated in the study and were distributed in perceptive and mnemonic experimental conditions. The effect of the instructions (intentional and incidental) on the representation of the distances among the objects of the scene was estimated using exponents of power function, based on the reproduction of the distances among the stimuli of the scene. The results revealed that judgments made under intentional instructions were more frequently based on strategies related to the location of the stimuli, whereas judgments originating from incidental instructions were based on strategies related to the name of the stimuli. It was observed that the intentional instruction facilitated a more accurate mental representation of the observed experimental configuration, enhancing participants' performance. PMID- 17120711 TI - Visual distance estimation in static compared to moving virtual scenes. AB - Visual motion is used to control direction and speed of self-motion and time-to contact with an obstacle. In earlier work, we found that human subjects can discriminate between the distances of different visually simulated self-motions in a virtual scene. Distance indication in terms of an exocentric interval adjustment task, however, revealed linear correlation between perceived and indicated distances but with a profound distance underestimation. One possible explanation for this underestimation is the perception of visual space in virtual environments. Humans perceive visual space in natural scenes as curved, and distances are increasingly underestimated with increasing distance from the observer. Such spatial compression may also exist in our virtual environment. We therefore surveyed perceived visual space in a static virtual scene. We asked observers to compare two horizontal depth intervals, similar to experiments performed in natural space. Subjects had to indicate the size of one depth interval relative to a second interval. Our observers perceived visual space in the virtual environment as compressed, similar to the perception found in natural scenes. However, the nonlinear depth function we found can not explain the observed distance underestimation of visual simulated self-motions in the same environment. PMID- 17120712 TI - Does energy expenditure affect the perception of egocentric distance? A failure to replicate experiment 1 of Proffitt, Stefanucci, Banton, and Epstein (2003). AB - In a series of recent studies, Proffitt and his colleagues have reported that the perceived distance to a target is influenced by the energy expenditure associated with any action, such as walking or throwing, for spanning the distance to the target. In particular, Proffitt, Stefanucci, Banton, and Epstein (2003) reported that wearing a heavy backpack caused verbal reports of distance to increase. We conducted a study to determine whether three responses dependent on perceived distance (verbal report of distance, blind walking, and estimates of object size) are influenced by the backpack manipulation. In two experiments, one involving a between-participants design and the other involving a within-participants design, we found that none of the three responses were influenced by the wearing of a heavy backpack. PMID- 17120713 TI - [Between good news and concern: how to face demographic aging?]. PMID- 17120714 TI - [Postprandial hypotension: an unclear clinical entity]. AB - Postprandial hypotension (PPH) is a clinical entity considered to affect above all elderly people with hypertension. It is equally common in diastolic heart failure, Parkinson's disease, diabetes mellitus and autonomic dysfunction. Diagnosis is based on a minimum of 20 mmHg drop of the systolic blood pressure oron a symptomatic systolic blood pressure decrease within 2 hours of the meal. Post-prandial dizziness, fatigue, syncope and falls must raise suspicion for this entity. Although more frequent than orthostatic hypotension, PPH is less searched for. Socio-economical repercussions associated to falls require a better screening of PPH in hospital and ambulatory conditions. PMID- 17120715 TI - [Drug-induced arterial hypertension]. AB - In a few clinical cases, controlling arterial hypertension is really very difficult even when using the appropriate treatment(s) at the optimal dosage. In such cases, it is crafty to research any possible drug interferences with the antihypertensive treatment. Two clinical cases will illustrate that arterial hypertension can be either induced or very difficult to control due to opposite physiopathological mechanisms of drugs or to pharmacokinetic interactions. A review of the main interacting drug regimens will complete this paper, attesting that drugs as various as NSAID, corticosteroids, erythropoietin, cyclosporine, antidepressants, beta blockers, carbamazepine, sympathomimetics, antihistamines, tramadol and (...) can induce arterial hypertension. PMID- 17120716 TI - [How to decide when a situation is too complex?]. AB - Health care professionals frequently face difficult clinical situations where no simple solution is available. In these situations, a pragmatic, structured ethical approach could be very helpful in defining ethical conflicts, systematically listing all possible options, and determining ethical justifications to their decisions. PMID- 17120717 TI - [Detection and coping of violence in care]. AB - Violence is an intrinsic phenomenon which deeply disturbs each human being. Violence can be either considered as a threat or as a source of energy. In fact violence is always driven by a vital force which interplays in any human relationship. Violence is a gradual process which is closely linked to the individual's personality, life history, pathology and reaction in confrontation to a precise situation within a determined context. Care givers have to be aware of the early symptoms of violence, identify its main causes to better cope with the patient's aggression and try to avoid catastrophic outcomes. PMID- 17120718 TI - [Anxiety in palliative care]. AB - In palliative care, the intensity and duration of anxiety as well as its consequences on the patient's daily activities can significantly decrease his quality of life. Anxiety that does not incapacitate the patient to the point of his being unable to communicate or perform his usual activities does not necessarily require drug treatment. The non pharmacological treatments of anxiety are presented in some detail. Prescription of anxiolytic drugs in renal or hepatic failure, as well as when oral intake or venous access are difficult, is briefly discussed. PMID- 17120719 TI - [Spirituality and elderly patients]. AB - Spirituality includes notions as diverse as the purpose and meaning of one's life, the feeling of internal peace and harmony. Several epidemiological studies have shown evidence of an association between spirituality and mental, physical, as well as functional health status. These evidences, in conjunction with the high incidence of spiritual distress observed in older patients, increased the interest toward improving the assessment and management of spiritual distress in these older patients. Several ongoing studies investigate the benefits to expect from such intervention. PMID- 17120720 TI - [Chronic lymphoid B-cell leukemia]. PMID- 17120721 TI - [Social inequalities and access to health care: consequences of the revision of the Swiss insurance law (art 64A)]. AB - The revision of the Swiss health insurance law (article 64A) excludes an increasing portion of the Swiss population from access to health care, and specifically to medications. From May to August 2006, 84 patients were referred to the University Hospital's primary care outpatient clinic to receive medications. These patients had a low socio-economic position, suffered from chronic diseases (88%) and had a high prevalence of psychiatric diseases (59%). This change in the Swiss insurance law mainly has an impact on the more fragile members of society and threatens the concept of equal access of the whole population to the national health care system. PMID- 17120722 TI - [As one puts on the white coat, one wonders...]. PMID- 17120723 TI - [Avian influenza: the European media under the eyes of Brussels]. PMID- 17120724 TI - [The new-found lesson of Dr. Destouches]. PMID- 17120726 TI - [International traffic of human cadaveric fragments stolen in the United States]. PMID- 17120725 TI - [Acomplia, an essential medication or a "comfort" product?]. PMID- 17120727 TI - [Stethoscope]. PMID- 17120728 TI - Vaccination with a recombinant chicken FGFR-1 bypasses immunological tolerance against self-FGFR-1 in mice. AB - The possibility that a recombinant protein vaccine based on xenogeneic homologous FGFR-1 of chicken induces production of autoantibodies against self-FGFR-1 in BALB/c mice was examined by using ELISA, Western blot analysis and ELISPOT assay respectively. Autoantibodies against mouse FGFR-1 were identified by Western blot analysis and ELISA. Compared with the two control groups, the number of APBCs, which were detected by ELISPOT assay, was significantly increased in the spleens of mice immunized with cFR1 (P < 0.05). IgG1 and IgG2b, which were detected by ELISA, were the major subclasses and were substantially increased in response to chicken FGFR-1 when compared with control group. The recombinant chicken FGFR-1 protein used as a vaccine can induce autoantibodies against self-FGFR-1 in mice and provide a basis for the active immunotherapy of tumor angiogenesis. PMID- 17120729 TI - Inhibition of PCNA antisense oligonucleotides mediated by liposome on mRNA expression and proliferation of h-RPE cells. AB - The proliferating cell nuclear antigen (PCNA) gene expression was blocked and retinal pigment epithelium (RPE) proliferation was inhibited by using antisense oligonucleotides (AS-ODN) mediated by liposome, to find a new genetic therapy of proliferative vitreoretinopathy (PVR). RPE cells cultured in vitro were transfected with synthetic fluorescence labled AS-ODN mediated by liposome Lipofectamine, and the intracellular distribution and persistence time of AS-ODN were dynamically observed. AS-ODN (0.07, 0.28 and 1.12 micro mol/L and sense oligonucleotides (S-ODN with the same concentrations as AS-ODN) mediated by liposome were delivered to the RPE cells cultured in vitro, and CPM values were measured by 3H-TdR incorporation assay and analyzed statistically by variance by comparison with blank control group. Expression of PCNA mRNA in RPE cells was detected by in situ hybridization after the treatment of different concentrations of PCNA AS-ODN and S-ODN, and the average optic density (AOD) was measured by image analysis system and was subjected to q-test and correlation analysis with CPM. Our results showed that AS-ODN mediated by liposome could quickly aggregate in cellular plasma and nuclei in 30 min and 6 h, and stayed for as long as 6 days. AS-ODN (0.28 and 1.12 micro mol/L) markedly suppressed proliferation of RPE cells in a dose-dependent manner with the difference being statistically significant (P < 0.05 and P < 0.01, repectively) as compared with blank control group. AOD was well correlated with CPM (r = 0.975). It is concluded that liposome could increase transfection efficiency of AS-ODN in RPE cells, and AS ODN could sequence-specifically suppress PCNA mRNA expression and proliferation of human RPE cells. PMID- 17120730 TI - Effect of cytokines secreted by human adipose stromal cells on endothelial cells. AB - To isolate and culture adipose stromal cells (ASCs), and study the effect of cytokines secreted by ASCs on endothelial cells, human adipose tissue was digested with collagenase type I solution and ASCs were derived by culture. The cells surface phenotype was examined by flow cytometry. ELISA was used to detect the secretion of VEGF, HGF, SDF-1 alpha and RT-PCR was employed to detect the expression of their mRNA. Then the ASC medium was utilized to culture human umbilical vein endothelial cells ECV304. Cells were counted by hemacytometer to determine the proliferation and Annexin V/ PI was employed for the examination of the apoptosis rate of ECV304. ASCs were derived by culture and expressed CD34, CD105 while they did not express CD31 or CD45. ASCs secreted cytokines such as VEGF, HGF and SDF-1 alpha so the ASC medium could stimulate proliferation and counteract apoptosis of endothelial cells (P < 0.05). Bcl-2 mRNA was also found to be up-regulated in the endothelial cells. It is concluded that ASCs can secrete cytokines and has significant effect on the proliferation of endothelial cells and apoptosis. PMID- 17120731 TI - Induction of silencing effect of Swedish mutant amyloid precursor protein by RNA interference. AB - Over-expression of APP and Swedish mutation could cause some familial early onset AD. In this study, a primary screening was conducted of effective small interference RNAs (siRNAs) targeted wild type APP (APPwt) and Swedish mutant APP (APPswe). One siRNA targeting APPwt and the other siRNA targeting APPswe were designed. All these siRNAs were endogenously expressed by siRNAs expressing plasmids. COS-7 cells were transiently co-transfected with APP-GFP recombinant plasmids and siRNA expression vector. The silencing effect of each siRNA was quantitatively assessed by the level of expression of green fluorescent protein (GFP). It was found that the siRNAs silenced APPwt and APPswe to different degrees. siRNA directed against APPswe was more effective in suppressing the expression of fusion gene of APPswe than that of APPwt. The silencing effect of siRNA directed against APPswe indicating allele-specific silencing property of the siRNAs. Therefore, siRNAs directed against APP play an important role both in the therapeutic study of Alzheimer disease and functional exploration ofAPP gene. PMID- 17120732 TI - Expression and significance of Bcl-2, Bax, Fas and caspase-3 in different phases of human hemangioma. AB - The relationship between Bcl-2, Bax, Fas, caspase-3 and development of hemangioma and the molecular mechanism was investigated. By using immunohistochemical S-P method, proliferating cell nuclear antigen was detected. According to the classification of Mulliken in combination with PCNA expression, 27 cases were identified as proliferating hemangioma and 22 cases as involutive hemangioma. Five normal skin tissues around the tumor tissue served as controls. By using immunohistochemical technique, the expression of Bcl-2, Bax, Fax and Caspase-3 was detected. The cells expressing Bcl-2, Bax, Fax and cappase-3 were identified as hemangioma endothelia by immunohistochemical staining of VIII factor. The average absorbance (A) and average positive area rate of Bcl-2, Bax, Fas and caspase-3 expression were measured by using HPIAS-2000 imaging analysis system. The results showed that the expression of Bcl-2 in the endothelia of proliferating hemangioma was significantly higher that in involutive degenerative hemangioma endothelia and vascular endothelia of normal skin tissue (P < 0.01). The expression of Bax, Fas and Caspase-3 in the endothelia of involutive hemangioma was obviously higher than in the endothelia of proliferating hemangioma and normal skin tissue (P < 0.01). The expression of BAx and Fas in endothelia of proliferating hemangioma was higher than in those of normal skin tissue (P < 0.05). It was suggested that Bcl-2, Bax, Fas and caspase-3 might be involved in the development and involution of hemangioma. Bcl-2 could promote the growth of hemangioma by inhibiting apoptosis of endothelia. Bax, Fas and caspase 3 promote the switch of hemangioma from proliferation to involution by inducing the apoptosis of hemangioma endothelia. PMID- 17120733 TI - The value of p63 and CK5/6 expression in the differential diagnosis of ductal lesions of breast. AB - In order to explore the value of p63, smoothmuscle actin (alpha-SMA) and cytokeratin 5/6 (CK5/6) in the differential diagnosis of ductal lesions of breast, 88 tissue specimens of ductal lesions of breast were collected and examined histologically by HE staining. By using immunohistochemistry, the expression of p63, alpha-SMA and CK5/6 was detected. The results showed that in 38 cases of benign breast lesions, the proliferating cells were all positive for p63 and alpha-SMA. In 19 cases of ductal carcinoma in situ (DCIS) and 7 cases of intraductal papillary carcinoma, alpha-SMA positive cells formed a layer of continuous embroider-shaped structure and the p63 positive cells formed a layer of evenly separated embroider-shaped structure around the ducts. There was no cross-reaction between p63 and interstitial myofibroblasts and vascular smooth muscle cells. In 38 cases of benign breast lesions, the positive rate of CK5/6 expression was 100%. In 5 cases of atypical ductal hyperplasia, there were few positive cells in the ducts. In 19 cases of CDIS, no tumor cells expressed CK5/6. In 19 cases of invasive ductal carcinoma, almost no CK5/6 was detectable. It was suggested that p63 could serve as a novel specific marker for the identification of breast myoepithelial cells. CK5/6 is of value in differentiating ductal proliferation of varying degrees, especially in the differentiation between cancerous and non-cancerous changes. Simultaneous detection of p63, CK5/6 and alpha-SMA can help increase the diagnostic accuracy of breast diseases. PMID- 17120734 TI - E sequence analysis of persistently infected mutant Japanese encephalitis virus strains. AB - A persistent infection model was established after human hepatoma cells infected by Japanese encephalitis viruses were subcultured for several times. Viral titers of mutant viruses in persistently infected cells were examined by plaque methods using BHK cells. Nucleotides of the E coding region of two wild and two mutant viruses were amplified by RT-PCR. PCR products were sequenced by ABI-PRSM 310 sequencing system. Compared to JaGAr-01 wild strains, four amino acids were replaced (E61Tyr --> Asp, E219His --> Tyr, E384Val --> Glu, E418Pro --> Ala) in the E sequence of JaGAr-01 persistently-infected mutant strains. Eleven amino acid replacement (E51Arg --> Ser, E61Tyr --> Asp, E83Lys --> Glu, E123Ser --> Arg, E209Arg --> Lys, E227Pro --> Ser, E276Asp --> Ser, E290Arg --> Lys, E387Lys -> Arg, E418Leu --> Pro, E454Arg --> Gly) was also noted when we compared the E sequence between persistently infected Nakayama and its wild strains. A lot of similarities of amino acid sequence between mutant strains JaGAr-01 and Nakayama were also noted. It was concluded that geno-variation existed in E region of mutant viruses and the mutant protein encoded by E region, especially the mutation of E61 (Tyr --> Asp) may contribute to the maintenance of the persistent infection of Japanese encephalitis virus. PMID- 17120735 TI - Characteristics of ex vivo expansion of endothelial progenitor cells. AB - The characteristics for the ex vivo expansion of the endothelial progenitor cells (EPCs) were explored. CD34+ cells were selected from umbilical cord blood mononuclear cells (MNC) by MiniMACS system, expanded under the same conditions as those for total MNC, coincubation of CD34+ and CD34- from the same donor for EPCs. In addition, the effects of vessel endothelial growth factor (VEGF) and passage on cell differentiation, expansion kinetics and apoptosis were examined. EPCs were determined and quantified by immunocytochemistry and flow cytometry. The results showed that both coculture of CD34+ and CD34- and total MNC led to a significant increase in the expansion of CD34+ cells as compared with CD34 enrichment (P < 0.05). There was a tendency toward decreased apoptosis in cultures when early passage was performed immediately after cord like structures appeared. VEGF had no significant effect on apoptosis (P > 0.05). These differentiated EPCs were positive for CD34+, von Willebrand factor (vWF), KDR, CD31 staining and phagocytized acetylated low-density lipoprotein (LDL). CD34+ cells accounted for (68.2 +/- 6.3)% of attaching (AT) cells at day 7 of culture. It was suggested the most efficient method to ex vivo expansion of EPCs was coculture of CD34+ and CD34- or total MNC. Early passage makes cell apoptosis rate decrease. VEGF had no significant effect on ex vivo expansion of EPCs. PMID- 17120736 TI - P53 regulation of leukemia cells with the blockage of MDM2 by antisense oligonucleotides. AB - The changes of expression and function of MDM2 and P53 by MDM2 specific antisense oligonucleotides were investigated in HL60 cells. Cells were divided into control group, AS group (MDM2 specific antisense oligonucleotides group), cisplatin group, and combined treatment group. FCM analysis and Western blot and RT-PCR were used to estimate apoptosis and the expression of MDM2 and P53. Our results showed that the transfection of MDM2 specific antisense oligonucleotides obviously inhibited MDM2 expression (P < 0.01) and increased the expression of P53 (P < 0.05). Apoptosis rate were reduced by MDM2 specific antisense oligonucletides and cisplatin (P < 0.01). It is concluded that MDM2 specific antisense oligonucletides can inhibit the expression of MDM2, induce the expression of P53 and increase the apoptosis of leukemia cells after chemotherapy. PMID- 17120737 TI - The dynamic change of TGF-beta1 in the myocardial remodeling of rat after myocardial infarction. AB - To observe the dynamic changes of the TGF-beta1 expressed in the infarct and non infarcted region of rat heart during the ventricular remodeling (day 3, 7, 28, 180), myocardial infarction rat model was made and relationship between the cytokine and indicator of myocardial remodeling was analyzed. After the detection of hemodynamic parameter was performed by the Powerlab devices, the size of myocardial infarction and the morphology change was detected by TTC and HE, respectively. The relative levels of mRNA of TGF- beta1, collagen type I, III, and fetal gene beta-MHC were detected by RT-PCR. The distribution of TGF- beta1 protein in the myocardium was detected by immunohistochemistry. The results showed that the size of infarction was higher than that of the sham operated groups in the infarcted group (44.5 +/- 0.5 vs 0). The difference in hemodynamic parameters between the infarcted group and sham operated group was significant (P < 0.01). HE staining showed that inflammatory cells were accumulated in the infarcted region at the beginning of the 3rd day, which lasted 4 weeks. Then, it decreased gradually. beta-MHC in the non-infarcted region rose from the 3rd day, reaching its peak at the 4th week, and it decreased gradually. The ratio of the collagen type I/III showed similar changes as compared with the sham operated groups (P < 0.01). And the relative mRNA levels in the non-infarcted group were significantly higher than that in the infarcted and sham operated group (P < 0.01) at day 180. Linear regression analysis indicated that the TGF-beta1 was positively correlated with the ventricular remodeling. It was concluded that the cytokine TGF-beta1 participates in the process of the myocardial remodeling, which could be a strategy in the interference of myocardial remodeling. PMID- 17120738 TI - Efficacy and safety of intravenous moxifloxacin versus cefoperazone with azithromycin in the treatment of community acquired pneumonia. AB - To compare the efficacy, safety, and tolerability of intravenous moxifloxacin with those of a commonly used empirical antibiotic regimen, cefoperazone and azithromycin in the treatment of community acquired pneumonia (CAP) in adult patients requiring initial parenteral therapy, 40 patients with CAP were divided into two groups, a moxifloxacin group (n = 20) and a control group (n = 20), which were treated for 7 to 14 days. The patients in the moxifloxacin group were intravenously given 400 mg of moxifloxacin (Avelox) once a day. Patients in the control group were administered 2.0 g of cefoperazone twice a day and azithromycin 0.5 g once a day. Clinical, bacteriological, and laboratory examinations were performed before the treatment, and at the end of the treatment. Our results showed that there was no significant difference in the clinical efficacy rate between two treatment groups at end of therapy (90% for moxifloxacin, 95% for cefoperazone plus azithromycin) (P > 0.05). The bacteriologic eradication rate at the end of treatment was 90% in the moxifloxacin group and 80% in the cefoperazone-plus-azithromycin group, whereas there was no significant difference between the two groups (P > 0.05). In addition, both drugs were well-tolerated in this trial, with the number of drug related adverse events being comparable. It is concluded that moxifloxacin is an effective and well-tolerated treatment for CAP and was equivalent to the commonly used empirical treatment of cefoperazone plus azithromycin. Moxifloxacin is likely to offer clinicians an alternative for reliable empirical CAP treatment in the face of increasing antibiotic resistance. PMID- 17120739 TI - Apoptosis signaling pathway in a subtotal nephrectomy rat model. AB - To investigate the role and mechanisms of apoptosis and apoptosis signaling pathway in 5/6 nephrectomy rat model (SN(x)), the mRNA and protein levels of caspase-3, -8, -9 and apoptosis were detected by in situ end labeling (TUNEL), immunohistochemistry, RT-PCR, Western-blotting 1, 2, 4, 8, 12, 16, 26 and 40 weeks after 5/6 nephrectomy rat model was made respectively. The rats in the model group developed glomerular sclerosis and renal interstitial fibrosis. The number of the apoptototic cells in glomeruli, renal tubule and renal interstitium was remarkably higher in the model group than that in the control group (P < 0.05, P < 0.01). Changes of mRNA and protein level of caspase-3, -8, -9 had the same tendency and was up-regulated wavily in the rat model compared with the control group (P < 0.05). Peaks in model appeared on the 4th and the 40th week respectively. The growth amplitude of caspase-9 was remarkably higher than that of caspase-8. It is concluded that the development of 5/6 nephrectomy rat model was correlated with the apoptosis of glomeruli, renal tubule and renal interstitium. Both of death receptor and mitochondria signaling pathways are involved in the process and the latter might play a primary role. PMID- 17120740 TI - Significance of nueroelectrophysiological tests in the early diagnosis of sub clinical neuropathay with diabetes mellitus. AB - To evaluate the application of nueroelectrophysiological tests in early diagnosis of sub-clinical neuropathy in diabetes mellitus (DM), The routine nerve conductive velocity (NCV), F-wave and sympathetic skin response (SSR) were detected in 27 patients with diabetes mellitus but without symptoms and signs of lesions of nerve system. Our results showed that 48.1%, 44.4%, 51.9% of the patients were found to have abnormal NCV, F-wave and SSR respectively. The abnormalities were mainly characterized by prolonged latency, reduced velocity and absence of wave-form. There were significant differences between the controls and the DM group (P < 0.05). Both the distal and proximal segments of nerves were affected and the distal lesions took place earlier than proximal ones and the changes in low extremities were more severe than those of upper extremities. F wave can be used as a sensitive indicator for the early diagnosis of peripheral neuropathy and it can help to detect the subclincial lesions. SSR can be used for the evaluation of functional status of autonomic nerves in DM patients. PMID- 17120741 TI - Naloxone or vagotomy does not influence centrally octreotide-induced inhibition of gastric acid secretion in rats. AB - To investigate the effect of preceding naloxone injection into the third cerebroventricle or acute subdiaphragmatic vagotomy on the gastric acid secretion inhibited by the somatostatin analogue octreotide given by intracerebroventricular (icv) injection. The third ventricles were cannulated in male Wistar rats anesthetized with sodium pentobarbital. One week later, acute gastric lumen perfusion was carried out. The gastric perfusion samples were collected every 10 min and were titrated by 0.01 mol/L NaOH to neuter. On the basis of subcutaneous injection of pentagastrin (G-5, 160 micro, g/kg), icv injection of physiological saline (group A, n = 20), icv injection of octreotide (0.05 micro g) (group B, n = 20), icv injection of naloxone (2.5 micro g)+octreotide (0.05 micro g) (group C, n = 20), acute subdiaphragmatic vagotomy+ icv injection of physiological saline (group D, n = 20), or acute subdiaphragmatic vagotomy+icv injection of octreotide (0.05 micro g) (group E, n = 20) were conducted. Before and after icv injection, 1-h total acid output (TAO) was determined and compared. The experimental data were expressed in change rate (%) of TAO. The change rates (%) of TAO were 4.60% in group A, -20.35% in group B, -18.06% in group C, 5.01% in group D and -21.59% in group E, respectively. Comparison of group B or C versus group A showed that P < 0.01 and comparison between the group E versus group D showed that P < 0.01. Whereas the differences between group C and group B, group E and group B were not statistically significant (P > 0.05 for all). The results indicate that the central inhibition of gastric acid secretion by octreotide may not be mediated by the endogenous opiate substance or its receptor and the peripheral pathway for icv injection of octreotide to suppress gastric acid secretion is via extra-vagus route. PMID- 17120742 TI - Protective effects of focal ischemic preconditioning and HSP70 expression on middle cerebral artery occlusion in rats. AB - To systematically evaluate the importance of protein synthesis in ischemic preconditioning (PC)-induced ischemic tolerance (IT), temporary middle cerebral artery occlusion (MCAO) by Longa (20 min) was used for PC (ischemic precondioning). Twenty-four hours of reperfusion was allowed after PC and before permanent MCAO to establish ischemic tolerance (IT) to compare with non-PC (sham operated) rats (n = 5 for each group). Infarct size and neurological deficits were measured 24 h after PMCAO. Samples of brain were taken for the determination of HSP70 expression by Western blot analysis. The effects of the protein synthesis inhibitor cycloheximide administered just before PC or administered long after PC but just before PMCAO on IT were also determined (n = 5 for each group). Our results showed that hemispheric infarct was significantly reduced (P < 0.01) only if PC was performed after 24 h, and PC significantly (P < 0.05) reduced neurological deficits (similar to reductions in infarct size). Cycloheximide eliminated ischemic PC-induced IT effects on bothbrain injury and neurological deficits if administered before PC but not if administered long after PC but before PMCAO. PC produced no brain injury but did increase HSP70 protein 24 h after PC. Cycloheximide eliminated that effect. The results suggest that PC is a powerful inducer of ischemic brain tolerance as reflected by the preservation of brain tissue and motor function. PC induces IT that is dependent on de novo protein synthesis. PMID- 17120743 TI - Cerebral ischemic tolerance induced by 3-nitropropionic acid is associated with increased expression of erythropoietin in rats. AB - To examine the changes in erythropoietin (Epo) protein and its mRNA expression in rat brain subjected to focal ischemia and possible mechanism of the preconditioning of mitochondrial toxin 3-nitropropionic acid (3-NPA), rats were administrated either vehicle or 3-NPA at a dose of 20 mg/kg, intraperitoneally (ip), 3 days prior to a 2-h middle cerebral artery occlusion followed by 24-h reperfusion. Infarct volumes were measured by using 2, 3, 5 triphenylte trazolinm chloride (TTC) staining, and Epo protein and its mRNA levels were assessed by immunohistochemistry and reverse transcriptase polymerase chain reaction (RT PCR), respectively. Our results showed that after reperfusion, Epo was found to be expressed extensively in the rat brain. It was most apparent in the basal nuclei and hippocampus, and was, to some extent, present in cortex. Preconditioning with 3-NPA caused a reduction in infarct volume. The expression of both Epo protein and mRNA increased significantly in the different brain areas in the 3-NPA pretreated group as compared with the non-pretreated ischemia model group. These results suggested that preconditioning with low dose 3-NPA could induce ischemic tolerance and neuro-protective effects by increasing the Epo expression in the ischemic and ischemia-related areas. PMID- 17120744 TI - Correlation between protein expression of PTEN in human pancreatic cancer and the proliferation, infiltration, metastasis and prognosis. AB - In order to investigate the correlation between protein expression of PTEN and the proliferation, infiltration, metastasis and prognosis in pancreatic cancer, immunohistochemical SP method was used to examine the protein expression of PTEN, PCNA, MVD, MMP-2, MMP-9 and TUNEL method to detect the levels of apoptosis of pancreatic cells in 41 pancreatic head cancers from regional pancreatectomy (RP) and 10 normal pancreatic tissues. The results showed that among 41 cases of pancreatic cancers, the positive staining of PTNE (39.02%) was significantly weaker than that in normal pancreatic tissues (P < 0.05). The levels of PCNA labeling index (LI), apoptotic index (AI), microvessel density (MVD), MMP-2 LI and MMP-9 LI were decreased gradually with the increase of the expression intensity of PTEN, and there was a significant difference in the above parameters among the patients having different expression levels of PTEN (P < 0.01 or P < 0.05). There was a negative correlation between the expression of PTEN and PCNA LI, MVD, MMP-2 LI, MMP-9 LI, and a positive correlation between AI and the expression of PTEN. The expression intensity of PTEN was correlated with the postoperative survival of the patients with pancreatic cancer (chi2 = 22.3400, P < 0.0001, RR = 2.030). It was suggested that the expression levels of PTEN protein were closely related with proliferation, infiltration and metastasis in human pancreatic cancer, and the expression of PTEN protein was one of the prognostic factors for pancreatic cancer following RP. PMID- 17120745 TI - Antisense oligonucleotide of hypoxia-inducible factor-1alpha suppresses growth and tumorigenicity of lung cancer cells A549. AB - To study the role and mechanisms of hypoxia-inducible factor-1alpha (HIF-1alpha) on the growth and tumorigenicity of lung cancer cells A549, the antisense oligonucleotide of HIF-1alpha was transfected to A549 cells. The effect of the antisense oligonucleotide on tumor growth in vitro and in vivo was evaluated by the growth rate suppression of A549 cells and subcutaneous implanted tumor in nude mice, and the effect on tumorigenicity was evaluated by the expression inhibition of angiogenic factors, the microvessel density (MVD) and vascular endothelial growth factor (VEGF) protein expression which were detected by immohistochemistry and western blot respectively. This study revealed that in vitro the growth rate of antisense oligonucleotide group was significantly decreased as compared with that of control group, sense oligonucleotide group and false-sense oligonucleotide group; in vivo the weight of implanted tumors in nude mice of antisense oligonucleotide group was 1.51 +/- 0.40 g, which was significantly lower than that of control group (2.79 +/- 0.33 g), sense oligonucleotide group (2.81 +/- 0.45 g) and false-sense oligonucleotide group (2.89 +/- 0.39 g) and the inhibitory rate was 47%. Both MVD and VEGF protein expression were significantly inhibited in antisense oligonucleotide group compared with those in other groups. These results indicated that antisense oligonucleotide of HIF-1alpha could inhibit lung cancer cells A549 growth in vitro and in vivo, and the mechanism may be due to the inhibition of vascular growth and VEGF protein expression. PMID- 17120746 TI - Culture and identification of monoclonal neural stem cells derived from cerebral cortex. AB - To isolate and culture the purified monoclonal neural stem cells from the cerebral cortex of new born mice, new-born mice cerebral cortex was isolated and dissociated to single-cell suspension by mechanical trituration. The dissociated single cells were cultured in serum-free medium. After the formation of neurospheres, single-cell clone culture was performed by limiting dilution and the proliferated single-cell clones were harvested for subculture. Immunocytochemistry was used to detect the specific marker of neuroepithelial stem cells (Nestin) of the primary and monoclonal neurospheres. In the differentiated cells we detected the specific antigen of NF-200 and GFAP. Our results showed that the primary neurospheres expressed Nestin antigen positively. By limiting dilution, we cultured the cell lines from single-cell clone and the monoclonal neurospheres expressed Nestin and had capabilities of self-renewal, proliferation and the potentiality of differentiation into neurons and glial cells. It is concluded that monoclonal neural stem cells which have the ability of proliferation and multi-directional differentiation can be isolated and cultured from the cerebral cortex of new-born mice by limiting dilution. PMID- 17120747 TI - Semi-skeletonized internal mammary grafts and phrenic nerve injury: cause-and effect analysis. AB - Phrenic nerve injury after cardiac surgery increases postoperative pulmonary complications. The purpose of this study was to analyze the causes and effects of phrenic nerve injury after cardiac surgery. Prospectively collected data on 2084 consecutive patients who underwent cardiac surgery from Jan. 1995 to Feb. 2002 were analyzed. Twenty-eight preoperative and operation related variables were subjected to logistic analysis with the end point being phrenic nerve injury. Then phrenic nerve injury and 6 perioperative morbidities were included in the analysis as variables to determine their independent predictive value for perioperative pulmonary morbidity. An identical approach was used to identify the independent risk factors for perioperative mortality. There were 53 phrenic nerve injuries (2.5%). There was no phrenic nerve injury in non-coronary surgery or coronary surgery using conduits other than the internal mammary artery. The independent risk factors for phrenic nerve injury were the use of internal mammary artery (Odds ratio (OR) = 14.5) and thepresence of chronic obstructive pulmonary disease (OR = 2.9). Phrenic nerve injury was an independent risk factor (OR = 8.1) for perioperative pulmonary morbidities but not for perioperative mortality. Use of semi-skeletonized internal mammary artery harvesting technique and drawing attention to possible vascular or mechanical causes of phrenic nerve injury may reduce its occurrence. Unilateral phrenic nerve injury, although rarely life-threatening, is an independent risk factor for postoperative respiratory complications. When harvesting internal mammary arteries, it should be kept in mind avoiding stretching, compromising, or inadvertently dissecting phrenic nerve is as important as avoiding damage of internal mammary artery itself. PMID- 17120748 TI - Effect of icariin on cyclic GMP levels and on the mRNA expression of cGMP-binding cGMP-specific phosphodiesterase (PDE5) in penile cavernosum. AB - To further investigate the mechanisms of action of icariin (ICA), we assessed the effects of ICA on the in vitro formation of cGMP and cAMP in isolated rabbit corpus cavernosum. Isolated segments of rabbit corpus cavernosum were exposed to increasing concentrations of ICA and the dose-dependent accumulation of cGMP and cAMP was determined in the tissues samples by means of 125I radioimmunoassay. Responses of the isolated tissues preparations to ICA were compared with those obtained with the reference compounds sildenafil (Sild). Furthermore, the effects of ICA on the mRNA expression of specific cGMP-binding phosphodiesterase type V (PDE5) in rat penis were also observed. After incubation with ICA for 6 h or 14 h respectively, the levels of PDE5 mRNA were examined by reverse transcriptase polymerase chain reaction (RT-PCR). The results showed that ICA increased cGMP concentrations directly (P < 0.05), but there was no significant effect on cAMP concentrations (P > 0.05). In the presence of sodium nitroprusside (SNP), a stimulatory agent of cGMP, both ICA and Sild increased cGMP concentrations with increasing dose (P < 0.01). Their EC50 was 4.62 (ICA) and 0.42 (Sild) micromol/L respectively. Under the same condition, ICA and Sild unaltered cAMP level significantly (P > 0.05). There were PDE5A1 and PDE5A2 mRNA expressions in rat corpus cavernosum with PDE5A2 being the dominant isoform. ICA could obviously inhibit these two isoforms mRNA expression in rat penis, and decrease PDE5A1 more pronouncedly (P < 0.01). The present study indicated that the aphrodisiac mechanisms of icariin involved the NO-cGMP signal transduction pathway, with increasing cGMP levels in the corpus cavernosum smooth muscle. The inhibitory effect of icariin on PDE5 mRNA expression, especially on PDE5A1, might account for its molecular mechanisms for its long-term activity. PMID- 17120749 TI - Up-regulation of HIF-1alpha and VEGF expression by elevated glucose concentration and hypoxia in cultured human retinal pigment epithelial cells. AB - In order to explore the effect of high glucose concentration and high glucose concentration with hypoxia on the production of hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF), human RPE cells were cultured in 5.56 mmol/L glucose (control group), 5.56 mmol/L glucose with 150 micro mol/L CoCl2 (hypoxic group), 25 mmol/L glucose (high glucose group) and 25 mmol/L glucose with 150 micro mol/L CoCl2 (combination group). RT-PCR was used to detect the expression of HIF-1alpha and VEGF mRNAs. Western blot analysis was used to measure the levels of HIF-1alpha and VEGF proteins. Although the small amount of HIF-1alpha protein was able to be detected in high glucose group but not in control group, there was no significant difference between the expression of HIF-1alpha mRNA of RPE cells in high glucose group and that of RPE cells in control group. As compared with RPE cells in control group, the mRNA expression and the protein synthesis of VEGF in high glucose group were up-regulated. As compared with RPE cells in hypoxic group, the expression of HIF-1alpha mRNA of RPE cells in combination group was not different, but the protein synthesis of HIF-1alpha, the mRNA expression and the protein synthesis of VEGF were more obviously up-regulated. In conclusion, high concentration glucose mainly influence the protein synthesis of HIF-1alpha of RPE cell, and HIF-1alpha protein is able to be accumulated in high concentration glucose. Under hypoxia, the HIF 1alpha protein induced by high concentration glucose is more stable, and the expression of VEGF is obviously increased. It is suggested that high concentration glucose may play a role in retinal neovascularization, especially at ischemia stage of diabetic retinopathy. PMID- 17120750 TI - CXCL12/CXCR4 expression in trophoblasts takes part in materno-fetal immune tolerance and vascular remodeling. AB - In this study, we investigated the expression of CXCL12 (SDF-1)/CXCR4 in trophoblasts and the role they play in the gestation. Immunochemistry was used to detect the expression of CXCR4 and CXCL12 in human villi and placenta. Highly purified extra-villous trophoblasts (EVTs) ere detected for CXCR4 and CXCL12 in vitro by immunocytochemistry. The chemotaxis of CXCL12 was tested in transwell and the chemotactic activity was quantitatively examined. It was suggested that both CXCR4 and CXCL12 were expressed in trophoblasts and were decreased with the gestation time P < 0.05). In a certain coverage, CXCL12 exhibited chemotactic activity which was positively correlated with its concentration [(r) = 0.68, P < 0.01], the maximum chemotactic index (CI) was 1.62 +/- 0.12. Our results suggest that interaction between CXCR4 and CXCL12 is involved in materno-fetal immunological tolerance in all three trimesters of gestation and contributes to the invasion of EVTs during pregnancy. PMID- 17120751 TI - Cyclosporine inhibits apoptosis in experimental murine xerophthalamia conjunctival epithelium. AB - This study examined the inhibitory effect of topical cyclosporine (CsA) treatment on conjunctiva epithelial apoptosis in a murine model of xerophthalamia. Dry eye was induced in 3 groups of C57BL6 mice by subcutaneous injection of scopolamine (t.i.d) and exposure to an air draft and low-humidity environment for 16 h each day for 12 days. The dry eye control group received no topical treatment; another group received 1 microL of 0.05% CsA topically (t.i.d, dry eye+CsA); and the third group received 1 microL of the castor oil vehicle of CsA topically (t.i.d, dry eye + vehicle). Normal mice were used as untreated controls. Twelve days later, the mice were killed, and their conjunctivas were excised. The number of the conjunctival goblet cells was counted in tissue sections stained with periodic acid Schiff (PAS) reagent. Their conjunctiva epithelium had been investigated by immuno-histochemical staining to detect the goblet cells and the expression of Caspase-3, Bax and bcl-2. Our results showed that compared with dry eye control and dry eye mice + vehicle groups, the number of conjunctival epithelial goblet cells was significantly greater in the untreated controls and dry eye mice receiving CsA (P < 0.01 for both groups). There was no significant difference in the number of conjunctival epithelial goblet cells between the dry eye control and dry eye+vehicle group. It was also true of the number of conjunctival epithelial goblet cells when comparison was made between the normal group and the dry eye+CsA group. Expressions of Caspas-3 and Bax were increased and ex-pression of bcl-2 was decreased in conjunctival epithelial cells in dry eye control and dry eye mice+vehicle groups. There was a significant positive correlation between goblet cell number and the number of cells that expressed bcl 2, and a negative correlation between goblet cells and Caspase-3 and Bax expression. It is concluded that the topical use of CsA could significantly reduce conjunctival epithelial apoptosis and protect goblet cell against the loss in experimental murine xerophathalamia. Inhibition of apoptosis appears to be a key mechanism responsible for the therapeutic effect of CsA on xerophthalamia. PMID- 17120752 TI - Correcting the corneal power measurements after myopic laser in situ keratomileusis. AB - To describe and evaluate a refraction-derived method and a clinically derived method to calculate the correct corneal power for intraocular lens (IOL) power calculations after laser in situ keratomileusis (LASIK) and to compare the results to the commonly used history-derived method. Retrospective analysis were conducted in consecutive case from clinical practice. For each patient, we established the pre-LASIK and post-LASIK spectacle refraction, the pre-LASIK (K(pre)) and post-LASIK K readings (K(post)). We then calculated the pre- and post-LASIK refraction at the cornmeal plane and the amount of correction obtained by the refraction surgery (deltaSEQco). The cases were divided into two groups. Group I was used to derive two formulas. The values obtained with the two methods were compared with the K by history-derived method (K(c.hd) in group II to validate the results. The K values calculated by using the refraction-derived method (K(c.rd)) and the K values calculated using the clinically derived method (K(c.cd)) correlated highly with K(c.hd). The correct corneal power for intraocular lens (IOL) power calculations after LASIK can use refraction-derived method and clinically derived method instead of history-derived method when some refractive parameters are not available. PMID- 17120753 TI - Comparative study of two vitreous humor sampling methods in rabbits. AB - To compare and evaluate two methodologies, entire-sampling and micro-sampling for the harvesting of vitreous humor, the vitreous humor of rabbits were sampled with the two methods respectively, and the concentrations of calcium, chlorine, potassium, sodium and phosphorus of the were measured. The results showed that the differences in the variance coefficient and two-eye concentrations of micro sampled specimens were less than those of the entire-sampled specimens. In the micro-sampling group, the concentrations of repeated micro-sampling showed no differences among different groups (P > 0.05) and the intra-ocular fluid dynamics did not have significant influence on post-mortem sampling. The sampling technique may affect the concentrations of specimen collected. Our study suggests that micro-sampling is less influenced by the human factor and is reliable, reproducible, and more suitable for forensic investigation. PMID- 17120754 TI - Effects of morphine, fentanyl and tramadol on human immune response. AB - Morphine has been reported to suppress human immune response. We aimed to observe the effects of morphine, fentanyl and tramadol on NF-kappa B and IL-2 from both laboratory and clinical perspective. Jurkat cells were incubated with ten times clinically relevant concentrations of morphine, fentanyl and tramadol before being stimulated with PMA. NF-kappa B binding activity and IL-2 levels were measured. In the clinical study, 150 consenting patients were randomized into 3 groups according to the analgesics used in them, namely, group morphine (M), group fentanyl (F) and group tramadol (T). IL-2 was measured preoperatively and 1, 3 and 24 h after operation. Consequently, NF-kappa B activation was suppressed by morphine and fentanyl but not by tramadol. IL-2 was significantly decreased by morphine and fentanyl but not by tramadol in vitro. In the PCA patients, IL-2 was decreased in group M and increased in group F postoperatively. Whereas in group T, IL-2 was unchanged 1 h after operation but was significantly elevated 3 and 24 h after operation. Our results showed that the inhibition of morphine on IL-2 was most probably related to its suppression on NF-kappa B. Fentanyl had different effects on human immune response in vitro and in vivo. Tramadol may have immune enhancing effect. PMID- 17120755 TI - Early reversible ischemia of femoral head epiphysis in piglets on gadolinium enhanced MRI: an experimental study. AB - The purpose of this study is to demonstrate if Gadolinium-enhanced MRI can detect early reversible ischemia of the femoral head epiphysis caused by hip hyper abduction in piglets. Between 3 and 6 h consistent hyper-abduction, gadolinium enhanced MRI was performed in 20 femoral heads of 10 piglets. After completion of MRI scan, the piglets were allowed to ambulate freely for 1 or 7 days and re imaged. The enhanced-MRI results of epiphyseal and physeal cartilage and the secondary center of ossification were observed. MRI appearances and histological findings were compared. On Gadolinium-enhanced MRI, decreased or absent enhancement was seen in 14 cartilaginous epiphyses of all 20 femoral heads. Reperfusion was completed in 10 of 14 femoral heads after one day of ambulation and in the rest 4 after 7 days of ambulation. Gadolinium-enhanced MRI can identify early ischemia and its reversal of the capital femoral epiphysis induced by hip hyper-abduction. PMID- 17120756 TI - Application of multislice spiral CT for guidance of insertion of thoracic spine pedicle screws: an in vitro study. AB - To investigate the value of the guidance of three dimensional (3-D) reconstruction of multi-slice spiral CT (MSCT) for the placement of pedicle screws, the 3-D anatomical data of the thoracic pedicles were measured by MSCT in two embalmed human cadaveric thoracic pedicles spines (T1-T10) to guide the insertion of pedicle screws. After pulling the screws out, the pathways were filled with contrast media. The PW, PH, TSA and SSA of developed pathways were measured on the CT images and they were also measured on the real objects by caliper and goniometer. Analysis of variance demonstrated that the difference between the CT scans and real objects had no statistical significance (P > 0.05). Moreover, the difference between pedicle axis and developed pathway also had no statistical significance (P > 0.05). The data obtained from 3-D reconstruction of MSCT demonstrated that individualized standards, are not only accurate but also helpful for the successful placement of pedicle screws. PMID- 17120757 TI - Magnetic resonance imaging of transplanted neural stem cells in Parkinson disease rats. AB - In this study we implanted magnetically labeled neural stem cells (NSCs) in PD rats and then monitored their survival and migration in the host brain by magnetic resonance imaging (MRI). The mesencephalic NSCs were obtained from the brain of SD rats. Superparamagnetic iron oxide (SPIO) was transferred to NSCs by Lipofectamine transfection. Eighteen PD lesioned rats were selected for transplantation by evaluation of their rotational behavior in response to amphetamine and randomly assigned to 3 groups, i.e., sham group, PBS group and NSCs transplanted group, with 6 rats in each group. MR scanning was performed at 1, 2, 4, 6, 8 and 10 week(s) following transplantation. At the meantime, rotational behavior was assessed in each group. Our results showed that SPIO particles were clearly visible with Prissian blue staining in neurospheres and cells derived from NSCs. The rotational behavior of the NSCs transplanted group was remarkably improved compared with that of sham group and PBS group (P < 0.05). In vivo MR tracking of NSCs showed that SPIO labeling led to a strong susceptibility change of signal 1 week after transplantation on T2 weighted images. And a large circular hypointense signal appeared in the transplanted area on T2* gradient echo images. Ten weeks following transplantation, the hypointense signal on T2 weighted and T2* gradient echo images was still displayed. It is concluded that SPIO particles could label NSCs effectively, and MRI detection of SPIO labeled cells is a promising method and novel approach to analyzing the NSCs following transplantation in the treatment of PD. PMID- 17120758 TI - Overview of the SLC26 family and associated diseases. AB - In the late 1990s the SLC26 family of anion exchangers emerged as the second, structurally distinct gene family capable of similar transport functions as the classical SLC4 or anion exchanger (AE) gene family. The observations leading to the characterization of the SLC26 family were firmly based on research on rare human diseases and aided by comparison to Caenorhabditis elegans. SLC26A1, or rat sulphate/anion transporter 1 (Sat1), was the first gene cloned in mammals, but not characterized in humans until the year 2000. Three rare recessive diseases in humans, namely diastrophic dysplasia (cartilage disorder resulting in growth retardation), congenital chloride diarrhoea (anion exchange disorder of the intestine) and Pendred syndrome (deafness with thyroid disorder) turned out to be caused by the highly related genes SLC26A2 (first called DTDST), SLC26A3 (first called CLD or DRA) and SLC26A4 (first called PDS), respectively. Subsequently, others and our laboratory cloned prestin, a cochlear motor protein gene (SLC26A5), a putative pancreatic anion transporter (SLC26A6), and SLC26A7 SLC26A11. Some SLC26 family members show highly specific tissue expression patterns, others are widely expressed. The SLC26 exchangers are capable of transporting, with different affinities, at least the chloride, iodide, sulfate, bicarbonate, hydroxyl, oxalate and formate anions, and have distinct anion specificity profiles. PMID- 17120759 TI - Individual characteristics of members of the SLC26 family in vertebrates and their homologues in insects. AB - The 10-member SLC26 gene family encodes anion exchangers of which SLC26A5 appears to be restricted to the outer hair cells of the inner ear. Here, the so-called prestin protein acts as a molecular motor, thought to be responsible for active mechanical amplification in the mammalian cochlea. We introduce special characteristics of SLC26A5 which may have relevance for other members of the family as well. As such, data point to a characteristic transcriptional control mechanism of which thyroid hormone surprisingly takes a role not only as an enhancer of expression, but also as a regulator of the subcellular redistribution of the prestin protein. Of significance for other members of the SLC26 family may be the observation that the failure of the subcellular redistribution of prestin protein prior to the onset of hearing leads to severe deficit of mature prestin function. Data will furthermore be argued in the context that prestin-related SLC26 proteins in the auditory organs of non-mammalian vertebrates and insects are widespread, possibly ancestral constituents of auditory organs and are likely to serve salient roles in mammals and across taxa. PMID- 17120760 TI - Sulfate transport by SLC26 transporters. AB - Sulfate is the fourth most abundant anion in human plasma that is essential for numerous physiological functions, including biotransformation of xenobiotics, steroids, non-steroidal anti-inflammatory drugs (NSAIDs), adrenergic stimulants/blockers and analgesics. Sulfate is also required for activation of many endogenous compounds (heparin, heparan sulfate, dermatan sulfate, bile acids) and utilized in the metabolism of neurotransmitters. Sulfation of structural components, including glycosaminoglycans and cerebroside sulfate, is essential for the maintenance of normal structure and function of tissues. Due to its hydrophilic nature, sulfate cannot readily cross the lipid bilayer of cells, thus plasma membrane proteins, known as sulfate transporters, are required for the movement of sulfate into/out of cells. Sulfate transporters can be divided into two distinct groups: Na+-dependent sulfate transporters belonging to the SLC13 gene family and Na+-independent sulfate transporters (antiporters, exchangers) belonging to SLC26 gene family. There are 11 members of the SLC26 family (including Sat1, DTDST, CLD, pendrin, prestin, cfex) whose structures and functions have been only partially characterized. In this presentation, the current information on the structures and functions of the sulfate transporters in the SLC26 gene family will be described and the issue that certain members of this family are unable to transport sulfate, will be addressed. PMID- 17120761 TI - Sugar transport by members of the SLC26 superfamily. AB - Two members of the SLC26 superfamily of transport proteins are expressed in the mammalian inner ear. These are pendrin (SLC26A4) and prestin (SLC26A5). Mutations in either lead to deafness but for different reasons. Prestin forms the molecular underpinning of the mechanical feedback step in cochlear amplification. The outer hair cell (OHC) motor basolateral membrane contains a high density of prestin (more than 10(7) copies per cell). OHCs can thus act as a laboratory to study this transporter. Before prestin was identified by a subtractive cDNA strategy, it was recognized that OHCs selectively transport fructose. Cylindrical OHCs swell reversibly and therefore shorten on exposure to iso-osmotic replacement of bath glucose with fructose. The uptake is saturable with Km = 16mM. We studied such transport in a heterologous expression system. Prestin expression levels, assayed by electrophysiology, were at least 30 times lower than in OHCs. Radioactive flux measurements show that fructose was transported into transfected cells. Measuring fluxes by cell swelling suggested that about 300 water molecules are co-transported per fructose, probably through the same salicylate-sensitive pore. Pendrin shares the same properties. The phenomenon reveals features of the extracellular surface of SLC26 while charge translocation reveals anion binding features of intracellular surface. PMID- 17120762 TI - SLC26A3 and congenital chloride diarrhoea. AB - Congenital chloride diarrhoea (CLD, OMIM214700) is a rare genetic disease caused by mutations in a plasma membrane protein, the solute-linked carrier family 26 member A3 (SLC26A3) protein, which encodes for an epithelial anion exchanger for Cl- and HCO3-. The main clinical symptom is a lifetime watery diarrhoea with a high Cl- content and low pH, causing dehydration and hypochloremic metabolic alkalosis. CLD may be fatal, if not adequately treated by substitution of NaCl, KCl and fluid lost in the faeces. Long-term prognosis is generally favourable, but complications such as renal disease, inflammatory bowel disease, hyperuricemia, inguinal hernias, spermatoceles and male subfertility are possible. The role of dysfunctional SLC26A3 in the pathogenesis of these complications is poorly known. Altogether 30 different mutations of the SLC26A3 gene are currently known among patients with CLD; the most common of them being the three founder mutations present among Finns (V317del), Polish (I675676ins) and Arabic (G187X) populations. Individual variation in the clinical picture of CLD is common, but not known to associate with the genotype. PMID- 17120763 TI - Expression, regulation and the role of SLC26 Cl-/HCO3- exchangers in kidney and gastrointestinal tract. AB - SLC26 isoforms are members of a large, conserved family of anion exchangers that display highly restricted and distinct tissue distribution. Cloning experiments have identified the existence of 10 SLC26 genes or isoforms (SLC26A1-11). The products of all, excepting SLC26A5 (prestin), function as anion exchangers with versatility with respect to transported anions. Modes of transport mediated by SLC26 members include the exchange of chloride for bicarbonate, hydroxyl, sulfate, formate, iodide, or oxalate with variable specificity. Several members of SLC26 family mediate chloride-bicarbonate exchange and display expression in a limited number of tissues including the gastrointestinal tract and/or kidney, with distinct subcellular (apical or basolateral) localization. These include SLC26A3 (DRA), SLC26A4 (pendrin), SLC26A6 (PAT1 or CFEX), SLC26A7 and SLC26A9. SLC26A3 and A9 are not expressed in the kidney but SLC26A4, A6 and A7 are. Genetically engineered null mice have highlighted the important role of two members of the SLC26 family, SLC26A4 and SLC26A6, in homeostatic function in kidney and/or intestine. In conjunction with expression studies, the evolving picture points to important roles for SLC26 family in chloride, bicarbonate, oxalate or sulfate transport and homeostasis in gastrointestinal tract, kidney and several other tissues. This review in particular focuses on the role and regulation of SLC26A6, A7 and A9 in the kidney and/or gastrointestinal tract. PMID- 17120764 TI - Anion exchangers in flux: functional differences between human and mouse SLC26A6 polypeptides. AB - The SLC26 anion transporter polypeptides exhibit considerably greater sequence diversity among near-species orthologues than is found among the SLC4 bicarbonate transporters, and among SLC26 transporters is most marked among SLC26A6 orthologues. This observation prompted systematic functional comparison in Xenopus oocytes of mouse Slc26a6 with several human SLC26A6 polypeptide variants. Mouse and human polypeptides exhibited similar rates of bidirectional [14C]oxalate flux, Cl-/HCO3- exchange, and Cl-/OH- exchange, and similar cAMP stimulation and enhancement of that stimulation by wild-type but not delta F508 CFTR. However, high rates of 36Cl- and 35S-sulfate transport by mouse Slc26a6 contrasted with low transport rates of the human proteins. The high 36Cl- transport phenotype cosegregated with the transmembrane domain of mouse Slc26a6 in chimera studies. Mouse Slc26a6 and human SLC26A6 each mediated electroneutral Cl-/HCO3- and Cl-/OH- exchange. But, whereas Cl-/oxalate exchange by mouse Slc26a6 was electrogenic, that mediated by human SLC26A6 appeared electroneutral. Oocyte expression of either mouse or human orthologue elicited currents that were pharmacologically distinct from the monovalent anion exchange activities measured in the same lots of oocytes. The human SLC26A6 polypeptide variants SLC26A6c and SLC26A6d were inactive in isotopic flux assays. Understanding of SLC26 transport mechanisms and pathophysiology will benefit from recognition of substantial differences in transport properties among orthologues. PMID- 17120765 TI - Physiology of electrogenic SLC26 paralogues. AB - SLC26 anion exchangers transport monovalent and divalent anions, with a diversity of anion specificity and stoichiometry. Our microelectrode studies indicate that several SLC26 members are electrogenic. We reported that Slc26a6 functions as a Cl-/formate, Cl-/oxalate, Cl-/OH- and electrogenic Cl-/nHCO3- exchanger. Recently, we have also confirmed that Slc26a7 does not behave as a Cl-/HCO3- exchanger but does function as an electrogenic anion conductance, perhaps a channel. We have also cloned murine Slc26a9, which is strongly expressed in the respiratory tract and stomach. Radioisotope uptakes in Xenopus oocytes indicate that Slc26a9 is a highly selective anion exchanger, transporting Cl- but neither formate, oxalate, nor SO42-. We also utilized electrophysiology to voltage clamp (VC) and/or measure intracellular pH (pHi), Cl- ([Cl-],) and Na+ ([Na+]i), in response to various ion replacements. Cl- removal in HCO3- depolarizes oocytes (to > +60mV), alkalinizes oocytes, and decreases aCl-i. Slc26a9 thus functions as an electrogenic nCl-/HCO3- exchanger, suggesting a role in pulmonary and gastric HCO3- secretion and/or CO2 transport. VC experiments revealed channel-like currents (>10 microA at -60mV and >80 microA at +60mV) mediated by Slc26a9 in the presence and absence of HCO3-. Our experiments and those of others continue to reveal additional characteristics and unique roles for this new class of electrogenic anion transporters. PMID- 17120766 TI - Role of SLC26-mediated Cl-/base exchange in proximal tubule NaCl transport. AB - The majority of the Na+ and Cl- filtered by the kidney is reabsorbed in the proximal tubule. In this nephron segment, a significant fraction of Cl- is transported via apical membrane Cl-/base exchange: Cl-/formate exchange in parallel with Na+/H+ exchange and H+/formate cotransport, and Cl-/oxalate exchange in parallel with oxalate/sulfate exchange and Na+/sulfate cotransport. Apical membrane Cl--OH- or Cl-/HCO3- exchange has also been observed. NHE3 mediates most if not all apical membrane Na+/H+ exchange in the proximal tubule. We evaluated SLC26 family members as candidates to mediate proximal tubule Cl /base exchange. We could not detect pendrin (SLC26A4) expression in the proximal tubule, and found no change in transtubular NaCl absorption in pendrin null mice. We did find expression of SLC26A6 (CFEX, PAT1) on the apical membrane of proximal tubule cells, and demonstrated that SLC26A6 is capable of mediating the Cl-/base exchange activities described to take place across the brush border membrane. Microperfusion studies on SLC26A6 null mice demonstrated that SLC26A6 is essential for oxalate-dependent NaCl absorption but does not contribute to baseline transport, suggesting it primarily mediates Cl-/oxalate exchange rather than Cl--OH- or Cl-/HCO3- exchange in the proximal tubule. Expression of SLC26A7 was also detected on the brush border membrane of proximal tubule cells. Finally, we demonstrated an essential role for the scaffolding protein PDZK1 in apical membrane expression of SLC26A6. PMID- 17120767 TI - SLC26 transporters and the inhibitory control of pancreatic ductal bicarbonate secretion. AB - SLC26 anion exchangers (probably SLC26A3 and SLC26A6) are expressed on the apical membrane of pancreatic duct cells and play a key role in HCO3- secretion; a process that is inhibited by the neuropeptide, substance P (SP). SP had no effect on basolateral HCO3- transporters in the duct cell or on CFTR Cl- channels, but inhibited a Cl- -dependent HCO3- efflux step on the apical membrane. In microperfused ducts, luminal H2DIDS (0.5mM) caused intracellular pH to alkalinize (consistent with inhibition of HCO3- efflux) and, like SP, inhibited HCO3- secretion. SP did not reduce HCO3- secretion further when H2DIDS was applied to the duct lumen, suggesting that SP and H2DIDS inhibit the same transporter on the apical membrane. As SLC26A6 is DIDS-sensitive, this isoform is the likely target for SP. The inhibitory effect of SP was mimicked by phorbol 12,13-dibutyrate (PDBu), an activator of protein kinase C (PKC). Moreover, bisindolylmaleimide, a blocker of PKC, relieved the inhibitory effect of both SP and PDBu on HCO3- secretion. Western blot analysis revealed that guinea pig pancreatic ducts express the alpha, beta1, delta, epsilon, eta, theta, zeta and mu isoforms o f PKC. We conclude that PKC is a negative regulator of SLC26 activity in pancreatic duct cells. PMID- 17120768 TI - Regulatory interaction between CFTR and the SLC26 transporters. AB - Most epithelia that express CFTR secrete fluid rich in HCO3- and poor in Cl- that is generated by a CFTR-dependent Cl- absorption and HCO3- secretion process that when aberrant leads to human diseases such as cystic fibrosis and congenital chloride diarrhoea. Epithelial Cl- absorption and HCO3- secretion require expression of CFTR and other Cl- and HCO3- transporters in the luminal membrane of the secreting cells. Recent advances in understanding this critical epithelial function revealed that the luminal Cl- and HCO3- transporters are members of the SLC26 family. Characterization of several members of the family reveals that all characterized thus far are electrogenic with an isoform specific Cl-/HCO3- transport stoichiometry. In vivo these transporters exist in a transporting complex with CFTR. The SLC26 transporters and CFTR are recruited to the complex by binding to scaffolds containing PDZ domains. Upon stimulation and PKA dependent phosphorylation of CFTR R domain, the R domain binds to the SLC26 transporter STAS domain. Interaction of the R and STAS domains results in a marked and mutual activation of CFTR and the SLC26 transporters. The significance of this mode of regulation to epithelial Cl- absorption and HCO3- secretion is obvious. PMID- 17120769 TI - Insights from a transgenic mouse model on the role of SLC26A2 in health and disease. AB - Mutations in the SLC26A2 cause a family of recessive chondrodysplasias that includes in order of decreasing severity achondrogenesis 1B, atelosteogenesis 2, diastrophic dysplasia and recessive multiple epiphyseal dysplasia. The gene encodes for a widely distributed sulfate/chloride antiporter of the cell membrane whose function is crucial for the uptake of inorganic sulfate that is needed for proteoglycan sulfation. To investigate the mechanisms leading to skeletal dysplasia, we generated a transgenic mouse with a mutation in Slc26a2 causing a partial loss of function of the sulfate transporter. Homozygous mutant mice were characterized by skeletal dysplasia with chondrocytes of irregular size, delay in the formation of the secondary ossification centre and osteoporosis of long bones. Impaired sulfate uptake was demonstrated in chondrocytes, osteoblasts and fibroblasts, but proteoglycan undersulfation was detected only in cartilage. The similarity with human diastrophic dysplasia makes this mouse a model to explore pathogenetic and therapeutic aspects of SLC26A2-related disorders. PMID- 17120770 TI - New insights into the role of pendrin (SLC26A4) in inner ear fluid homeostasis. AB - For over 100 years after the first description of the disorder, the molecular pathology underlying the deafness and thyroid pathology in Pendred syndrome (PS) remained unknown. In 1997, early progress towards understanding the molecular basis of the disorder was made when we identified the PS gene and found it to belong to the SLC26 family of anion transporters. The realization that an anion transporter was responsible for these clinical features soon highlighted a potential role for pendrin in thyroid hormone biosynthesis. The role of pendrin in deafness, however, remained unclear. Our determination of its expression pattern in the inner ear along with the development of a mouse with a targeted disruption of the Slc26a4 gene has revealed that Slc26a4 is expressed in areas of the endolymphatic compartment known to play a role in endolymph reabsorption and that absence of this protein leads to a profound prenatal endolymphatic hydrops and destruction of many of the epithelial cells surrounding the scala media. The precise mechanisms underlying endolymph reabsorption in the inner ear are not yet known; these studies, however, provide some of the groundwork for allowing the future delineation of these processes. PMID- 17120771 TI - The renal physiology of pendrin (SLC26A4) and its role in hypertension. AB - SLC26A4 (pendrin, PDS) is a Na+-independent, Cl-/HCO3-/OH- exchanger that is expressed in the apical regions of type B and non-A, non-B intercalated cells within the cortical collecting duct (CCD), the connecting tubule and the distal convoluted tubule where it mediates HCO3- secretion and Cl- absorption. SLC26A4 is upregulated with aldosterone analogues and with Cl- restriction. While under basal conditions no renal abnormalities are observed in mice and humans with genetic disruption of SLC26A4 (Pendred syndrome), differences become apparent under conditions wherein the transporter is stimulated. Following treatment with aldosterone analogues, e.g. deoxycorticosterone pivalate (DOCP), weight gain and hypertension are observed in Slc26a4+/+ but not in Slc26a4-/- mice. During dietary NaCl restriction, a model in which serum aldosterone is appropriately increased, urinary volume and urinary excretion of Cl- are greater in Slc26a4-/- than in wild-type mice which results in apparent vascular volume contraction in Slc26a4-/- mice. Moreover, during NaCl restriction or following DOCP treatment, Slc26a4-/- mice have a higher serum HCO3- than wild type mice from an impaired ability to excrete OH- equivalents. In conclusion, SLC26A4 regulates blood pressure and arterial pH, likely by participating in the renal regulation of net acid and Cl- excretion. PMID- 17120772 TI - Interaction of prestin (SLC26A5) with monovalent intracellular anions. AB - Outer hair cells (OHCs) of the mammalian cochlea are equipped with a specific form of cellular motility that is driven by changes of the membrane potential. This electromotility is a membrane-based process generated by the membrane protein prestin (SLC26A5). Current models suggest that prestin undergoes a force generating conformational transition upon changes of the membrane potential. The voltage dependence of prestin needs to be mediated by a charged particle within the protein, a 'voltage sensor', that can move through the membrane electrical field to trigger these conformational rearrangements. Indeed, voltage sensor translocation can be measured as electrical charge transfer. Here, we review and extend data indicating that charge movement by prestin and consequently electromotility depend on the presence of small monovalent anions such as chloride and bicarbonate at the cytoplasmic side of the membrane. The voltage dependence of prestin varies with concentration and species of the anion present, consistent with a partial translocation of the anion through the membrane. Thus anions may act as extrinsic voltage sensors. These conclusions suggest that charge movement and subsequent conformational rearrangements may relate to anion transport by other SLC26 members. Insights into molecular properties of prestin may provide clues to common mechanisms of anion transport by SLC26 proteins. PMID- 17120773 TI - "Stem cells into liver"--basic research and potential clinical applications. PMID- 17120774 TI - Mesenchymal stem cells increase self-renewal of small intestinal epithelium and accelerate structural recovery after radiation injury. AB - Patients who undergo pelvic or abdominal radiotherapy may develop side effects that can be life threatening. Tissue complications caused by radiation-induced stem cell depletion may result in structural and functional alterations of the gastrointestinal (GI) tract. Stem cell therapy using mesenchymal stem cells (MSC) is a promising approach for replenishment of the depleted stem cell compartment during radiotherapy. There is little information on the therapeutic potential of MSC in injured-GI tract following radiation exposure. In this study, we addressed the ability of MSC to support the structural regeneration of the small intestine after abdominal irradiation. We isolated MSC from human bone marrow and human mesenchymal stem cells (hMSC) were transplanted into immunotolerent NOD/SCID mice with a dose of 5.10(6) cells via the systemic route. Using a model of radiation induced intestinal injury, we studied the link between damage, hMSC engraftment and the capacity of hMSC to sustain structural recovery. Tissue injury was assessed by histological analysis. hMSC engraftment in tissues was quantified by PCR assay. Following abdominal irradiation, the histological analysis of small intestinal structure confirms the presence of partial and transient (three days) mucosal atrophy. PCR analysis evidences a low but significant hMSC implantation in small intestine (0.17%) but also at all the sites of local irradiation (kidney, stomach and spleen). Finally, in presence of hMSC, the small intestinal structure is already recovered at three days after abdominal radiation exposure. We show a structural recovery accompanied by an increase of small intestinal villus height, three and fifteen days following abdominal radiation exposure. In this study, we show that radiation-induced small intestinal injury may play a role in the recruitment of MSC for the improvement of tissue recovery. This work supports, the use of MSC infusion to repair damaged GI tract in patients subjected to radiotherapy. MSC therapy to avoid extended intestinal crypt sterilization is a promising approach to diminish healthy tissue alterations during the course of pelvic radiotherapy. PMID- 17120775 TI - Optimizing viral and non-viral gene transfer methods for genetic modification of porcine mesenchymal stem cells. AB - INTRODUCTION: Mesenchymal stem cells (MSCs) provide an excellent source of pluripotent progenitor cells for tissue-engineering applications due to their proliferation capacity and differentiation potential. Genetic modification of MSCs with genes encoding tissue-specific growth factors and cytokines can induce and maintain lineage-specific differentiation. Due to anatomical and physiological similarities to humans, porcine research models have been proven valuable for the preclinical testing of tissue engineering protocols in large animals. The aim of this study was to evaluate optimized viral and non-viral ex vivo gene delivery systems with respect to gene transfer efficiency, maintenance of transgene expression, and safety issues using primary porcine MSCs as target cells. MATERIALS AND METHODS: MSCs were purified from bone marrow aspirates from the proximal tibiae of four 3-month-old Danish landrace pigs by Ficoll step gradient separation and polystyrene adherence technique. Vectors expressing enhanced green fluorescent protein (eGFP) and human bone morphogenetic protein-2 (BMP-2) were transferred to the cells by different non-viral methods and by use of recombinant adeno-associated virus (rAAV)-mediated and retroviral gene delivery. Each method for gene delivery was optimized. Gene transfer efficiency was compared on the basis of eGFP expression as assessed by fluorescence microscopy and fluorescence-activated flow cytometry. BMP-2 gene expression and osteogenic differentiation were evaluated by realtime quantitative RT-PCR and histochemical detection of alkaline phosphatase activity, respectively. RESULTS: Non-viral gene delivery methods resulted in transient eGFP expression by less than 2% of the cells. Using high titer rAAV-based vector up to 90% of the cells were transiently transduced. The efficiency of rAAV-mediated gene delivery was proportional to the rAAV vector titer applied. Retroviral gene delivery resulted in long-term transgene expression of porcine MSCs. A 26-fold increase in percentage of eGFP expressing cells (1.7%+/-0.2% versus 44.1% +/-5.0%, mean +/ SD) and a 68-fold increase in mean fluorescence intensity (327.4+/-56.6 versus 4.8+/-1.3) was observed by centrifugation of retroviral particles onto the target cell layer. Porcine MSCs that were BMP-2 transduced by optimized retroviral gene delivery demonstrated a significant increase in BMP-2 gene expression and showed increased osteogenic differentiation. Retrovirally transduced porcine MSCs were furthermore tested free of replication-competent viruses. DISCUSSION: The non viral gene transfer methods applied were significantly less efficient compared to the viral methods tested. However, due to advantages with respect to safety issues and ease of handling, improvement of non-viral gene delivery to primary MSCs deserves further attention. The high efficiency of rAAV-mediated gene delivery observed at high titers can be explained by the ability of rAAV vector to transduce nondividing cells and by its tropism towards porcine MSCs. rAAV mediated gene delivery resulted in transient transgene expression due to lack of stable AAV genome integration. MLV-mediated retroviral gene delivery can be considered a safe method for long-term transgene expression by porcine MSCs, and is therefore particularly attractive for advanced tissue engineering strategies requiring extended transgene expression. PMID- 17120776 TI - Transplantation of bone marrow stromal cells for treatment of central nervous system diseases. PMID- 17120777 TI - Chondrocyte signaling and artificial matrices for articular cartilage engineering. AB - Chondrocytes depend on their environment to aid in their expression of appropriate proteins. It has been found that the interaction of integrin receptors with chondrocytes effects the production of extracellular molecules such as type II collagen and aggrecan. Additionally, the presence of growth factors such as IGF-1, TGF-beta1 and BMP-7 induce various signaling pathways that also aid in transducing phenotypic expressions by chondrocytes. Natural and synthetic polymers have been used to act as a scaffold for chondrocytes. The production of extracellular matrix proteins by chondrocytes has been studied. As tissue engineers, it is advantageous to explore the possibility of how altering biomaterial properties affect the signaling cascades by activation of receptors and transduction through the cytoplasm. This vital information will be able to aid in the future of engineering an appropriate biomaterial that can incorporate chondrocytes to act as a scaffold for articular cartilage. PMID- 17120778 TI - Osteoinduction with COLLOSS, COLLOSS E, and GFm. AB - This study provided data relevant to three major goals. It confirmed that both COLLOSS and COLLOSS E contain osteo- and chondro-inductive BMPs as shown by their ability to produce new bone in an ectopic location in rats. Second, based on the area of bone produced in standardized implant sections by osteoinductive growth factors in GFm, COLLOSS , and COLLOSS E and their respective collagenous carrier matrices, the study showed that COLLOSS was 0.1, and COLLOSS E 0.3 time as potent as 10 microg of GFm. Finally, the study showed that ordinary and accelerated endochondral bone formation were more frequent in response to GFm than to COLLOSS and COLLOSS E, whereas membranous bone formation was more frequent in response to COLLOSS E than to COLLOSS or GFm. PMID- 17120779 TI - Biglycan is a positive modulator of BMP-2 induced osteoblast differentiation. PMID- 17120780 TI - Use of neopterin as a bone marrow hematopoietic and stromal cell growth factor in tissue-engineered devices. AB - The in vitro response of early haematopoietic progenitors or stem cells (CD34+)- common for myeloid (granulocyte, eosinophil, megakaryocyte) and marrow stromal cell lineages, to neopterin, exogenously added to the liquid mouse bone marrow cultures, at doses 12.5-25 microg/ml culture medium, has been studied. The results obtained show a significant stimulation of common--myeloid and stromal/ mesenchymal progenitor cell proliferation and differentiation, as early as 24h to the 96h after the in vitro treatment with neopterin. On day 4 of cultivation the granulocyte/macrophageal proliferation and differentiation has been attenuated giving place to the marrow stromal/mesenchymal cell growth and differentiation. A functional role of neopterin as hematopoietic growth factor--essential for the proliferation and differentiation of bone marrow common (hematopoietic and stromal) progenitors is not yet clear and remains to be elucidated. The in vitro and ex vivo applying of neopterin--alone or in specific combinations with other cytokines (e.g. FGF-2) for the induction of marrow stromal/mesenchymal cell proliferation and differentiation, merits further investigations with regards to its future use in regenerative medicine. The results provide a theoretical basis for the application of neopterin in tissue-engineered devices: incorporated into biodegradable polymer microparticles (with encapsulated early bone marrow progenitors and other special supplements), it could be experimentally applied for fast and easy induction of endothelial, osteoblastic/osteogenic, neuronal and other cell lineage differentiation as well as for improving tissue trophical processes and reparative microenvironment. PMID- 17120781 TI - Injectable synthetic extracellular matrices for tissue engineering and repair. PMID- 17120782 TI - Temporal changes in peg hydrogel structure influence human mesenchymal stem cell proliferation and matrix mineralization. PMID- 17120783 TI - Novel biophysical techniques for investigating long-term cell adhesion dynamics on biomaterial surfaces. AB - Cell adhesion on biomaterial surface is crucial for the regeneration and function of clinically viable cell and tissues. In turn, the cellular phenotypes, following the mechanochemical transduction of adherent cells on biomaterials, are directly correlated to the biophysical responses of cells. However, the lack of an integrated bio-analytical system for probing the cell-substrate interface poses significant obstacles to understanding the behavior of cells on biomaterial surface. We have developed a novel method, based on the principle of confocal reflectance interference contrast microscopy (C-RICM) that has enabled us to study the biomechanical deformation of cells on biomaterial surfaces. In this article, we would like to describe our recent development of the C-RICM system that integrates a confocal fluorescence microscope, phase contrast microscope and GFP expression system. We shall demonstrate the system by determining the adhesion contact kinetics, initial deformation rate, cytoskeleton structures of adherent cells on extracellular matrices (e.g., collagen and fibronectin) and biodegradable polymer (e.g., poly(lactic acid)) during long-term culture. We shall demonstrate that this unique approach could provide valuable biophysical information necessary for designing optimized biomaterial surfaces for cell/tissue regeneration applications. PMID- 17120784 TI - Evaluation of various types of scaffold for tissue engineered intervertebral disc. PMID- 17120785 TI - Physicochemical characterization of photopolymerizable PLGA blends. PMID- 17120786 TI - Porous tantalum trabecular metal scaffolds in combination with a novel marrow processing technique to replace autograft. AB - INTRODUCTION: Interbody fusion requires a structural member to carry load while the autograft or osteoinductive agent stimulates bone formation. In the present study, we evaluated the potential use of extracted nucleated cells from bone marrow mixed in hyaluronic acid gel as an osteoinductive agent, in comparison to Collagraft loaded with nucleated cells or rhBMP-2 in the porous tantalum ring, in an anterior lumbar interbody fusion (ALIF) in pigs. METHODS: Four 3-month-old female Danish landrace pigs were employed in the current study. Bone marrow was collected by means of aspiration, from the medullary cavity of the proximal tibia. The nucleated cells were isolated with a Ficoll step gradient centrifugation. The cell adhered rate after 24 hours of cultivation and ALP activity in the osteogenic medium following 4 days of cultivation were measured. Cell numbers in the porous tantalum discs were assessed by CyQuant measurement, and fluorescent live/dead cell staining in the porous tantalum discs was performed after periods of 24 hours and 7 days of cultivation. The nucleated cells mixed in hyaluronic acid gel were cultivated on slides for 24 hours, 7 days and 21 days. The live/dead cell staining, ALP staining or osteocalcin staining, were performed. A porous tantalum ring was loaded with nucleated cells in hyaluronic acid gel or packed with Collagraft strips also with nucleated cells or rhBMP-2. Immediately after preparation, one of three implants was inserted into L2-3, L4-5 or L6-7 respectively. The pigs were killed 3 months postoperatively. The lumbar spine specimens were prepared for histological evaluation. RESULTS: The adhered rate, of the nucleated cells, was 2.26+/-0.56%. ALP activity was no different in the osteogenic culture compared to DMEM/10%FBS alone. Cell numbers and live/dead cells showed no difference in the porous tantalum discs. Histological appearance showed that nucleated cells mixed with hyaluronic acid gel, had more mature bone in the central hole of the porous tantalum ring, compared to Collagraft strips with nucleated cells or rhBMP-2. Bone volume fraction did not differ within the three porous tantalum rings; however, more marrow space in the central hole of the porous tantalum ring was present when nucleated cells mixed with hyaluronic acid gel (57.4%) compared to Collagraft strips with rhBMP-2 (29.7%). CONCLUSION: In the current study, we demonstrate that nucleated cells, which were isolated from bone marrow intraoperatively, could be used to replace autograft if nucleated cells mixed with hyaluronic acid gel or with Collagraft strips packed into the porous tantalum ring in the pig ALIF model. PMID- 17120787 TI - Preparation of sponge using porcine small intesinal submucosa and their applications as a scaffold and a wound dressing. PMID- 17120788 TI - Modulation of cell differentiation in bone tissue engineering constructs cultured in a bioreactor. AB - In summary, many factors can influence the osteoblastic differentiation of marrow stromal cells when cultivated on three-dimensional tissue engineering scaffolds. In creating ideal bone tissue engineering constructs consisting of a combination of a scaffold, cells, and bioactive factors; a flow perfusion bioreactor is a much more suitable culture environment than static culture in well plates. The bioreactor eliminates mass transport limitations to the scaffold interior and provides mechanical stimulation to the seeded cells through fluid shear. Scaffold properties such as pore size impact cell differentiation, especially in flow perfusion culture. In addition, the bone-like extracellular matrix created by the in vitro culture of marrow stromal cells on porous scaffolds creates an osteoinductive environment for the differentiation of other marrow stromal cell populations. Therefore, bone tissue engineering constructs created by in vitro culture have excellent potential for bone regeneration applications in the clinic. However, more work is required to optimize this tissue engineering strategy. A biodegradable material with mechanical integrity similar to native bone and degradation properties similar to the rate of bone formation would be a more ideal scaffold material. It is also yet unclear what the optimum scaffold pore size and amount of in vitro generated extracellular matrix are to maximize bone formation. Finally, better characterization of the flow patterns within the flow perfusion bioreactor is needed to better understand the relationship between fluid shear and cell differentiation for creation of the ideal scaffold/culture combination. PMID- 17120789 TI - Bioreactors for tissues of the musculoskeletal system. AB - Muskuloskeletal tissue includes bone, cartilage, ligament, skeletal muscle and tendons. These tissues malfunction either due to a natural injury, trauma, or a disorder. In all cases natural regeneration needs to be enhanced by medication and, in many instances, by surgery. Surgical techniques are limited to suturing, autografts or allografts. Tissue engineering stems from the challenge presented by the limited resources for natural implants and the ineffectiveness of previous curing techniques. The challenge in tissue engineering resides in the design of a functional bioreactor that would: (1) house the engineered construct under sterile conditions; and (2) provide the appropriate stimuli that would result in a neotissue with biochemical and biomechanical properties comparable to in situ tissue. The various types and designs of bioreactors for the regeneration of musculoskeletal tissue, including spinner flask, rotating wall vessel, flow perfusion, and mechanical loading devices are presented in this paper. PMID- 17120790 TI - Non-invasive monitoring of tissue-engineered pancreatic constructs by NMR techniques. PMID- 17120791 TI - From molecules to matrix: construction and evaluation of molecularly defined bioscaffolds. AB - In this chapter, we describe the fundamental aspects of the preparation of molecularly-defined scaffolds for soft tissue engineering, including the tissue response to the scaffolds after implantation. In particular, scaffolds prepared from insoluble type I collagen fibres, soluble type II collagen fibres, insoluble elastin fibres, glycosaminoglycans (GAGs) and growth factors are discussed. The general strategy is to prepare tailor-made "smart" biomaterials which will create a specific microenvironment thus enabling cells to generate new tissues. As an initial step, all biomolecules used were purified to homogeneity. Next, porous scaffolds were prepared using freezing and lyophilisation, and these scaffolds were crosslinked using carbodiimides. Crosslinking resulted in mechanically stronger scaffolds and allowed the covalent incorporation of GAGs. Scaffold characteristics were controlled to prepare tailor-made scaffolds by varying e.g. collagen to elastin ratio, freezing rate, degree of crosslinking, and GAGs attachment. The tissue response to scaffolds was evaluated following subcutaneous implantations in rats. Crosslinked scaffolds maintained their integrity and supported the formation of new extracellular matrix. Collagen-GAG scaffolds loaded with basic fibroblast growth factor significantly enhanced neovascularisation and tissue remodelling. Animal studies of two potential applications of these scaffolds were discussed in more detail, i.e. for bladder and cartilage regeneration. PMID- 17120792 TI - Age-related differences in articular cartilage wound healing: a potential role for transforming growth factor beta1 in adult cartilage repair. AB - Objective of this study was to investigate the early wound healing reactions of immature and mature articular cartilage on experimental wound healing in the New Zealand White rabbit. The proliferation potential and glycosaminoglycan production of isolated chondrocytes of these animals was studied in an alginate culture system. A band of tissue with death chondrocytes was observed at wound edges of immature articular cartilage, whereas mature cartilage showed a significant smaller amount of dead chondrocytes. A general increase in TGFbeta1, FGF2 and IGF1 was observed throughout cartilage tissue with the exception of lesion edges. The observed immunonegative area appeared to correlate with the observed cell death in lesion edges. Repair in immature cartilage was indicated by chondrocyte proliferation in clusters and a decrease in defect size. No repair response was observed in mature articular cartilage defects. The alginate culture experiment demonstrated a higher proliferation rate of immature chondrocytes. Addition of recombinant TGFbeta1 increased proliferation rate and GAG production of mature chondrocytes. We were not able to further stimulate immature chondrocytes. These results indicate that TGFbeta1 addition may contribute to induce cartilage repair responses in mature cartilage as observed in immature, developing cartilage. PMID- 17120793 TI - Intrinsic versus extrinsic vascularization in tissue engineering. PMID- 17120794 TI - Predictive value of in vitro and in vivo assays in bone and cartilage repair- what do they really tell us about the clinical performance? PMID- 17120795 TI - Engineered tissues: the regulatory path from concept to market. PMID- 17120796 TI - Fibrin in tissue engineering. PMID- 17120797 TI - Ectopic bone induction by equine bone protein extract. AB - Demineralized bone matrix from horse has been reported to be osteoinductive. However, its performance was inferior to autogenous bone graft in terms of new bone formation. In the present experiment, an equine bone protein extract-COLLOSS E was investigated for its osteoinductivity in a rat model. At the mean time, carboxymethyl-cellulose (CMC) was tested as a potential carrier for the protein extract. 18 male Wistar rats (8 weeks) were employed in the experiment. Each rat was implanted randomly with the 2 of the following implants, one on each side of the abdominal muscle. 1) COLLOSS E lyophilisate. 2) PEEK ring holder. 3) 3% or 10% CMC .in gel or lyophilized form 4) COLLOSS E lyophilisate with 3% CMC, implanted as gel or in lyophilized form. 5) COLLOSS E suspension with 10% CMC, implanted as gel or in lyophilized form. The rats were followed up for 21 days. After termination, samples were subjected to macroscopic examination, plain radiograph, micro-CT and histological evaluations. The results showed that PEEK ring or CMC alone could not induce ectopic bone formation. COLLOSS E lyophilisate has a slightly higher (6 out of 7) positive bone formation rate over COLLOSS E/3% CMC (3 out of 5, both gel and lyophilized form), however, the difference is non significant (p=0.36, Fisher's exact test). 10% CMC with COLLOSS E did not show ectopic bone formation when implanted as gel form (0/8), while 1 positive bone formation was found when implanted as the lyophilized form (1/4). Bone tissue volume ranged from 0 mm(3) to 23.1mm(3) for COLLOSS-E lyophilisate alone and 0 to 29.7mm(3) for COLLOSS E/3%CMC (gel or lyophilized form). We concluded that equine bone protein extract has the ability to induce ectopic bone formation in the rat model. CMC could be a potential carrier, however, further studies are needed to verify the proportion and efficacy. PMID- 17120798 TI - Adipose tissue induction in vivo. AB - Engineering adipogenic tissue in vivo requires the concomitant induction of angiogenesis to generate a stable long-term three-dimensional construct. Histioconductive tissue engineering strategies have been used. The disadvantage of using biodegradable scaffolds is a delayed angiogenic induction resulting in ischemic necrosis of the central cell population in the scaffold. We evaluated an histioinductive approach for adipose tissue engineering by combining essential key components for adipogenic induction: (1) a precursor cell source, (2) a vascular pedicle, (3) a supportive matrix, and (4) a chamber to preserve space for the new tissue to develop. We observed concomitant adipogenic and angiogenic induction after 6 weeks in three-dimensional adipose tissue constructs. PMID- 17120799 TI - Ocular tissue engineering. PMID- 17120800 TI - Molecular mechanism of osteochondroprogenitor fate determination during bone formation. AB - Osteoblasts and chondrocytes, which derive from a common mesenchymal precursor (osteochondroprogenitor), are involved in bone formation and remodeling in vivo. Determination of osteochondroprogenitor fate is under the control of complex hormonal and local factors converging onto a series of temporospatial dependent transcription regulators. Sox9, together with L-Sox5 and Sox6, of the Sox family is required for chondrogenic differentiation commitment, while Runx2/Cbfa 1, a member of runt family and Osterix/Osx, a novel zinc finger-containing transcription factor play a pivotal role in osteoblast differentiation decision and hypertrophic chondrocyte maturation. Recent in vitro and in vivo evidence suggests beta-catenin, a transcriptional activator in the canonical Wnt pathway, can act as a determinant factor for controlling chondrocyte and osteoblast differentiation. Here we focus on several intensively studied transcription factors and Wnt/beta-catenin signal molecules to illustrate the regulatory mechanism in directing commitment between osteoblast and chondrocyte, which will eventually allow us to properly manipulate the mesenchymal progenitor cell differentiation on bone and regeneration of cartilage tissue engineering. PMID- 17120801 TI - Mapping sound to meaning: connections between learning about sounds and learning about words. PMID- 17120802 TI - A developmental intergroup theory of social stereotypes and prejudice. AB - Developmental intergroup theory specifies the mechanisms and rules that govern the processes by which children single out groups as targets of stereotyping and prejudice, and by which children learn and construct both the characteristics (i.e., stereotypes) and affective responses (i.e., prejudices) that are associated with these groups in their culture. Specifically, we argue that children have a drive to understand their world, and that this drive is manifested in their tendency to classify natural and non-natural stimuli into categories, and to search the environment for cues about which of the great number of potential bases for categorization are important. The first step in the process of stereotype and prejudice formation is, therefore, the establishment of the psychological salience of some particular set of dimensions. Four factors are hypothesized to affect the establishment of the psychological salience of person attributes: (1) perceptual discriminability of social groups, (2) proportional group size, (3) explicit labeling and use of social groups, and (4) implicit use of social groups. We argue that person characteristics that are perceptually discriminable are more likely than other characteristics to become the basis of stereotyping, but that perceptual discriminability alone is insufficient to trigger psychological salience. Thus, for example, young children's ability to detect race or gender does not mean that these distinctions will inevitably become the bases of stereotypes and prejudice. Instead, for perceptually salient groups to become psychologically salient, one or more additional circumstances must hold, including being characterized by minority status, by adults' use of different labels for different groups, by adults using group divisions functionally, or by segregation. After a particular characteristic that may be used to differentiate among individuals becomes salient, we propose that children who have the ability to sort consistently will then categorize newly encountered individuals along this dimension. The act of categorization then triggers the process of social stereotyping and prejudice formation. Four factors are hypothesized to have an impact on the processes of forming stereotypes and prejudice. These include: (1) essentialism, (2) ingroup bias, (3) explicit attributions to social groups, and (4) group-attribute covariation. As noted throughout this chapter, there has been relatively little developmental work on many of the processes outlined here. Although findings from our own programs of research are consistent with the role of factors we have identified in the theory (e.g., the role of minority status, segregation, labeling and functional use of groups have all been shown to influence children's evaluations and beliefs about social groups), far more extensive research is needed. In addition to testing the reliability and generalizability of past findings to other samples, other research laboratories, and other experimentally manipulated groups, future work must move these theoretical models into the laboratory of the real world. If the tenets of developmental intergroup theory are correct, there would be many implications for social, educational, and legal policies related to social groups. We noted, for example, ways in which race and gender are made psychologically salient (e.g., the use of labels; segregated conditions). Importantly, factors such as these are largely under societal control. That is, institutions and individuals can choose to routinely label and use some particular category within children's environments or not. It is a violation of federal law, for example, for public school teachers to ask the children in their classrooms to line up at the door by race. In contrast, no federal or state law prohibits teachers from organizing their classrooms by sex. Should such laws be enacted? There can also be social controls on various forms of social segregation. Is it within individual children's rights to affiliate only with same-sex or same-race individuals? Is it acceptable for children and adolescents to exclude peers from their games, play, study groups, or other cliques on the basis of gender, race, age, or ethnicity? Finally, social institutions such as schools offer potential opportunities for intervention programs. What, if any, programs should be offered or required? Should curricula explicitly discuss social stereotyping and prejudice? Should children be taught negative information about people with whom they share some characteristic to reduce ingroup favoritism? Our hope is that developmental intergroup theory will ultimately prove valuable not only for understanding the development of social stereotypes and prejudices in children, but also for guiding social interventions that can ultimately prevent the development of stereotypes and prejudices in individuals and society. PMID- 17120803 TI - Income poverty, poverty co-factors, and the adjustment of children in elementary school. AB - Since 1990, there have been great advances in how developmental researchers construct poverty. These advances are important because they may help inform social policy at many levels and help frame how American culture constructs poverty for children, both symbolically and in the opportunities children and families get to escape from poverty. Historically, developmental perspectives have embodied social address and main effects models, snapshot views of poverty effects at single points in time, and a rather narrow focus on income as the symbolic marker of the ecology of disadvantage. More recent views, in contrast, emphasize the diverse circumstances of disadvantaged families and diverse outcomes of disadvantaged children, the multiple sources of risk and the multiple determinants of poor outcomes for these children, dynamic aspects of that ecology, and change as well as continuity in outcome trajectories. The advances also consist of more powerful frames for understanding the ecology of disadvantage and the risk it poses for child outcomes. Most developmental researchers still tend to frame causal variables ultimately in terms of the dichotomy between social causation and social selection views, with a primary emphasis on the former. In part, this framing has reflected limitations of sample size and design, because the theoretical and empirical power of reciprocal selection models is clear (Kim et al., 2003). The conceptual advances that prompt such models include widespread acknowledgement of third variable problems in interpreting effects, of the clear need for multivariate approaches, and the need to pursue mechanisms and moderators of the relations between causal candidates and child outcomes. In the context of these advances, one of the core goals of our research program has been to construct robust representations of environmental adversity for disadvantaged families. Most of our research focuses on contextual co-factors at a family level (e.g., maternal relationship instability), which either have not been described by many researchers or have been described in a way that does not fit the ecology of disadvantage (e.g., marital status). We found that income poverty, key contextual co-factors, and endogenous child attributes tend to show independent and selective associations with child academic competence and externalizing behavior, and that co-factor effects tend to be direct rather than mediated by harsh parenting, tend to have effects that are episodic and concurrent, and are easily- and well-represented by multiple risk indexes that bear powerful relations to child problem behaviors. A second core goal has been to better understand the developmental construction of poor outcomes for disadvantaged children, which requires consideration of dynamic aspects of the ecology and the potential importance of the timing of risk experiences. We found that family instability and change in environmental circumstances predict increases in problem behaviors, and that dose of adversity seems to matter for some variables if it is recent, and not for other variables. Through person-centered research, we also are beginning to understand some factors that seem to underlie the convergence of adjustment problems over grade in school. Many of our co-factor findings and many of our developmental findings seem both complex and double-edged. One edge is that they encourage a certain pessimism in showing how environmental adversity progressively constructs poor outcomes for disadvantaged children in school. Overall, for instance, we saw more problems and more multi-dimensional problems in fifth grade than in first grade, and the impact of environmental change was mostly negative. The other edge, however, is more positive in reflecting the possibility of discontinuity in child adjustment problems associated with positive changes in family circumstances. Findings for minimal persistence and for the strength of recent and concurrent effects argue that the possibility of self-righting and emergent competence in school is robust through the fifth grade even for the most problematic disadvantaged children. PMID- 17120804 TI - I thought she knew that would hurt my feelings: developing psychological knowledge and moral thinking. PMID- 17120805 TI - Home range and the development of children's way finding. PMID- 17120806 TI - The development and neural bases of facial emotion recognition. PMID- 17120807 TI - Children's suggestibility: characteristics and mechanisms. PMID- 17120808 TI - The emergence and basis of endogenous attention in infancy and early childhood. PMID- 17120809 TI - The probabilistic epigenesis of knowledge. PMID- 17120810 TI - Optimization via experimental design of an SPE-HPLC-UV-fluorescence method for the determination of valsartan and its metabolite in human plasma samples. AB - A chemometric approach was applied for the optimization of the extraction and separation of the antihypertensive drug valsartan and its metabolite valeryl-4 hydroxy-valsartan from human plasma samples. Due to the high number of experimental and response variables to be studied, fractional factorial design (FFD) and central composite design (CCD) were used to optimize the HPLC-UV fluorescence method. First, the significant variables were chosen with the help of FFD; then, a CCD was run to obtain the optimal values for the significant variables. The measured responses were the corrected areas of the two analytes and the resolution between the chromatographic peaks. Separation of valsartan, its metabolite valeryl-4-hydroxy-valsartan and candesartan M1, used as internal standard, was made using an Atlantis dC18 100 mm x 3.9 mm id, 100 angstroms, 3 microm chromatographic column. The mobile phase was run in gradient elution mode and consisted of ACN with 0.025% TFA and a 5 mM phosphate buffer with 0.025% TFA at pH 2.5. The initial percentage of ACN was 32% with a stepness of 4.5%/min to reach the 50%. A flow rate of 1.30 mL/min was applied throughout the chromatographic run, and the column temperature was kept to 40+/-0.2 degrees C. In the SPE procedure, experimental design was also used in order at achieve a maximum recovery percentage and extracts free from plasma interferences. The extraction procedure for spiked human plasma samples was carried out using C8 cartridges, phosphate buffer (pH 2, 60 mM) as conditioning agent, a washing step with methanol-phosphate buffer (40:60 v/v), a drying step of 8 min, and diethyl ether as eluent. The SPE-HPLC-UV-fluorescence method developed allowed the separation and quantitation of valsartan and its metabolite from human plasma samples with an adequate resolution and a total analysis time of 1 h. PMID- 17120811 TI - RP-HPLC prefractionation and its application in expressional proteomics analysis of an in vitro viral infection model. AB - Prefractionation of complex protein mixtures is an efficient method for increasing the separation power of 2-DE. RP-HPLC has been successfully utilized as a prefractionation method prior to 2-DE. Here we describe the optimization of an efficient RP-HPLC method for prefractionation of baby hamster kidney cell solubilized proteins. A step gradient elution of acetonitrile was optimized and collected fractions were further examined by SDS-PAGE and 2-DE. By utilizing this method an effective increase in separation power of 2-DE is accomplished. Moreover, we describe the application of this method to expressional proteome analysis of a virally infected cell model. PMID- 17120812 TI - Determination of 6-gingerol in ginger (Zingiber officinale) using high performance thin-layer chromatography. AB - A sensitive and accurate High-Performance TLC (HPTLC) method has been developed to determine the quantity of 6-gingerol in rhizomes of Zingiber officinale (family: Zingiberaceae), commonly known as ginger. Methanol extracts of rhizomes from three different sources were used for HPTLC, n-hexane, and diethyl ether (40:60 v/v) as the mobile phase. The Rf of 6-gingerol was found to be 0.40. The calibration plot was linear in the range of 250-1200 ng of 6-gingerol and the correlation coefficient of 0.9997 was indicative of good linear dependence of peak area on concentration. The mean quantity of 6-gingerol was found to be 60.44+/-2.53 mg/g of ginger extract. The method permits reliable quantification of 6-gingerol and good resolution and separation of 6-gingerol from other constituents of ginger. To study the accuracy and precision of the method, recovery studies were performed by the method of standard addition. Recovery values from 99.79 to 99.84% showed the excellent reliability and reproducibility of the method. The proposed HPTLC method for quantitative monitoring of 6 gingerol in ginger can be used for routine quality testing of ginger extracts. PMID- 17120813 TI - High performance liquid chromatography for the determination of glucosamine sulfate in human plasma after derivatization with 9-fluorenylmethyl chloroformate. AB - In this study, we developed a simple, rapid, sensitive, and reliable method for the determination of glucosamine sulfate in human plasma, which was based on derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) followed by reverse phase HPLC-FLD. For the first time, FMOC-Cl was introduced into derivatization of glucosamine sulfate in human plasma. The amino groups of glucosamine sulfate and vertilmicin sulfate (the internal standard) were trapped with FMOC-Cl to form glucosamine-FMOC-Cl and vertilmicin-FMOC-Cl adducts, which can be very suitable for HPLC-FLD. Precipitation of plasma proteins by acetonitrile was followed by vortex mixing and centrifugation. Chromatographic separation was performed on a C18 column (DIAMONSIL 150 x 4 mm id, 5 microm) with a mobile phase gradient consisting of acetonitrile and water at a flow-rate of 1 mL/min. The retention times of glucosamine-FMOC-Cl and vertilmicin-FMOC-Cl adducts were 8.9 and 21.2 min, respectively. This method was shown to be selective and sensitive for glucosamine sulfate. The limit of detection was 15 ng/mL for glucosamine sulfate in plasma and the linear range was 0.1-10 mg/mL in plasma with a correlation coefficient (r) of 0.9999. The relative standard deviations (RSDs) of intra-day and inter-day assays were 5.2-8.1% and 6.1- 8.5%, respectively. Extraction recoveries of glucosamine sulfate in plasma were greater than 90%. The validated method was successfully applied to the determination of glucosamine sulfate in human plasma samples. PMID- 17120814 TI - Development and validation of a reversed-phase HPLC method for monitoring of synthetic reactions during the manufacture of a key intermediate of an anti hypertensive drug. AB - A reversed-phase high-performance liquid-chromatographic method for monitoring of reactions involved in process development of a key intermediate of antihypertensive drugs, e.g, doxazosin mesylate, prazosin, alfuzosin, terazosin, etc., has been developed and validated. The HPLC profiles of impurities of 4 amino-2-chloro-6,7-dimethoxyquinazoline were used as fingerprints to follow the synthetic procedures in the manufacturing unit. The separation was accomplished on an Inertsil ODS-3 column with isocratic elution using acetonitrile-ammonium acetate (10 mM; pH 4.0; 50:50 v/v) as mobile phase and a photodiode array detector set at 240 nm at ambient temperature. The method was validated with respect to accuracy, precision, linearity, and limits of detection and quantification. The method could detect the impurities at a level of 0.01 to 0.20 microg/mL and it was found to be suitable not only for monitoring of reactions but also for quality assurance of 4-amino-2-chloro-6,7-dimethoxyquinazoline. PMID- 17120815 TI - Molecular imprinting of natural flavonoid antioxidants: application in solid phase extraction for the sample pretreatment of natural products prior to HPLC analysis. AB - As shown in the past years, SPE based on molecularly imprinted polymers (MIPs) may provide significant enhancement of selectivity in sample preparation and analyte preconcentration. The objective of this work was the fabrication of MIPs for the specific adsorption of rutin and quercetin. The two flavonoids were used as the template molecules for the preparation of MIP phases in a self-assembly (noncovalent) approach. The produced MIPs were validated with regard to the imprinting efficiency as media for LC and SPE. The retention behavior of several flavonoid compounds was studied using as stationary phases imprinted, control nonimprinted polymers, and commercial silica-based materials. MIPs were applied as materials for the selective SPE and preconcentration of the flavonoids from white and red wine, orange juice, and tea. The collected fractions were analyzed by high-pressure LC. MIP-SPE facilitated specific analyte isolation and effective sample clean-up. The results show that molecularly imprinted SPE can be a useful tool for the simple, selective, and cost-effective pretreatment of samples containing natural antioxidants. PMID- 17120816 TI - Determination of amphetamines in human urine by liquid chromatography with fluorimetric detection using a solid-phase extraction procedure. AB - A precise and feasible HPLC method has been developed for the analysis of amphetamine (AMPH), methamphetamine (MAMPH) and methylenedioxymethamphetamine (MDMA, ecstasy) in human urine. A chromatographic run on a C8 Genesis (150 mm x 4.6 mm, 5 microm) column maintained at 30 degrees C lasts about 17 min, using a mobile phase composed of ACN (12%) and a pH 2.5 phosphate buffer (88%) containing 0.3% triethylamine. Mirtazapine was used as the internal standard. Good linearity was found in the 100-2000 ng/mL concentration range for AMPH and MAMPH and in the 12-2000 ng/mL concentration range for MDMA. The pretreatment of urine samples was carried out by means of a careful SPE procedure on C2 cartridges. The extraction yields were very satisfactory for all analytes, with average values greater than 97%. The leading conditions allowed the determination of AMPH, MAMPH and MDMA with satisfactory precision and accuracy. The method has been successfully applied to the determination of the analytes in urine of AMPH users. PMID- 17120817 TI - Determination of multiclass pesticide residues in apple juice by gas chromatography-mass spectrometry with large-volume injection. AB - This study presents two GC-MS SIM methods, in combination with large-volume injection programmed-temperature vaporization (LVI-PTV) injection, for the determination of 141 pesticide residues in apple juice. The sample was extracted with ACN, and coextractives were removed with primary/secondary amine sorbent. ACN extract (20 microL) was injected into a PTV injection port in solvent vent mode, and the pesticides were determined by GC-MS using retention time locking software. Deuterium-labeled pesticides (surrogate standards) were used for analytical quality control. In the validation experiments, pesticides recoveries were found to be 70-121% with RSDs of 4.6-21% (n = 6). PMID- 17120818 TI - Low cost, robust, in-house hardware for heart cutting two-dimensional gas chromatography. AB - Natural materials are so complex that no single column can separate all of the components. Heart cutting 2-D GC (GC-GC) using a Deans switch provides maximum separation, but the requisite gas flow configurations have earned a reputation for being fiddly and time consuming to set up and tune, unless electronic gas controls and costly software are employed. A design for a vented Deans switch is presented that can be assembled in-house from standard commercial components. A vent to atmosphere replaces the balancing resistor, which solves the problem of balancing the pressures and flows, and requires no compensation for changes in gas viscosity and back-pressure during temperature programming. First and second dimension columns can both be run at optimum flow rates, even if they are of different diameters. Analyte detectability is preserved by cutting only the target fraction from the first column and transferring the whole of it to the second column. Cryotrapping the selected fraction allows cuts of any width, and transfer of analytes to the second column as sharp bands. I have applied the vented Deans switch-cold trap to the identification of flavour and fragrance compounds from South African plants and foods; cuts are very repeatable, and the detectability of trace components is enhanced. Its capabilities are demonstrated by examples from analyses of essential oils and flavour extracts. PMID- 17120819 TI - Comprehensive two-dimensional gas chromatographic analysis of commercial lemon flavored beverages. AB - Fresh lemon juice and lemon-flavored beverages were analyzed by using comprehensive 2-D GC (GC x GC) with flame-ionization detection, with a nonpolar polar column combination. A low-alcohol, ready-to-drink (RTD) beverage was also analyzed as fresh, and after deterioration for 12 days at 50 degrees C. Identification of some of the components in the 2-D plots was performed by comparison of peak positions of authentic standards and comparison with 1-D GC. However, without the aid of GC x GC-mass spectral data, only 24 components were identified; a large number of components remained unassigned. In some soft drinks obtained in the market, components indicative of deterioration, such as p methylacetophenone and p-cymen-8-ol were already present in the products. In contrast, even upon heat challenge, a low-alcohol RTD beverage did not generate deterioration products of citral, such as p-methylacetophenone and the intermediates, p-menth-2-ene-1,8-diols. This was apparently related to the fact that the original formulation contained only a minute amount of the citral ingredient. PMID- 17120820 TI - Determination of the 2,3-pentadienedioic acid enantiomer interconversion energy barrier 1. Classical kinetic approach. AB - A classical kinetic method was used to determine the energy barrier for the inter conversion of 2,3-pentadienedioic acid enantiomers. Each individual enantiomer was isolated by collecting the appropriate peaks from the HPLC enantiomeric separation, of racemic 2,3-pentadienedioic acid. The isolated enantiomers were racemized at 22 degrees C using various interconversion times. The ratio of enantiomers in each reaction solution was determined by HPLC at 22 degrees C. The corresponding peak areas of the enantiomers and the interconversion times obtained from the HPLC chromatograms were used to calculate both the interconversion rate constants describing (+)--> (-) and (-) --> (+) interconversions as well as the energy barriers. It was confirmed that the interconversion of 2,3-pentadienedioic acid enantiomers is a firstorder kinetic reaction. Both semiempirical and ab initio methods were used to explore the mechanism of the interconversion of 2,3-pentadienedioic acid enantiomers, and to calculate the interconversion energy barrier. Comparison of the interconversion energy barriers found by the ab initio method (deltaG# = 110.7 kJ/mol) and by classical kinetics in the mobile phase solution at 22 degrees C (delta Gapp = 93.9+/-0.2 kJ/mol) shows a difference which may be attributed to the different conditions assumed in the theoretical calculation (i.e., a gaseous state) and the actual experimental conditions (i. e., liquid solution) and a possible catalytic effect of the solution composition. PMID- 17120821 TI - Injector-internal thermal desorption from edible oils performed by programmed temperature vaporizing (PTV) injection. AB - Injector-internal thermal desorption is a promising technique for the analysis of a wide range of food components (e.g., flavors) or food contaminants (e.g., solvent residues, pesticides, or migrants from packaging materials) in edible oils and fats or fatty food extracts. Separation from the fatty matrix occurs during injection. Using programmed temperature vaporizing (PTV) injection, the oily sample or sample extract was deposited on a small pack of glass wool from which the components of interest were evaporated and transferred into the column in splitless mode, leaving behind the bulk of the matrix. Towards the end of the analysis, the oil was removed by heating out the injector and backflushing the precolumn. The optimization dealt with the gas supply configuration enabling backflush, the injector temperature program (sample deposition, desorption, and heating out), separation of the sample liquid from the syringe needle and positioning it on a support, deactivation of the support surface, holding the plug of fused silica wool by a steel wire, and the analytical sequence maintaining adsorptivity at the desorption site low. It was performed for a mixture of poly(vinyl chloride) (PVC) plasticizers in oil or fatty food. Using MS in SIM, the detection limit was below 0.1 mg/kg for plasticizers forming single peaks and 1 mg/kg for mixtures like diisodecyl phthalate. For plasticizers, RSDs of the concentrations were below 10%; for the slip agents, oleamide and erucamide, it was 12%. The method of incorporating PTV injection was used for about one year for determining the migration from the gaskets of lids for glass jars into oily foods. PMID- 17120822 TI - Studies on thermionic ionisation detection in comprehensive two-dimensional gas chromatography. AB - This study explores the application of specific thermionic ionisation detection in comprehensive 2-D GC (GC x GC) and represents the first report of GC x GC with nitrogen phosphorus detection (GC x GC-NPD). Of particular interest is the performance of the NPD with respect to peak parameters of asymmetry and sensitivity. Since GC x GC produces much narrower peaks than obtained with fast GC (e.g. 100 ms vs. <1 s) the effect of detector response time and any lack of symmetry arising from the detection step is important if peak separation (resolution) is to be maintained. It was observed that detector gas flows had a significant impact on peak asymmetry and peak magnitude, and that optimisation of the detector was critical, particularly for complex sample analysis by GC x GC NPD. Peak asymmetries ranging from As = 1.8 to 8.0 were observed under different conditions of detector gas flows. Comparison of GC x GC-NPD with GC x GC-flame ionisation detection (FID) showed the former to be approximately 20 times more sensitive for the detection of nitrogen-containing methoxypyrazines analytes, and GC x GC-NPD had a larger linear detection range compared to GC x GC-FID. Furthermore, comparison of GC x GC-NPD and GC x GC-TOFMS chromatograms for the analysis of coffee head-space demonstrated the benefits of selective detection, ultimately realised in a comparatively simplified contour plot. PMID- 17120823 TI - High-capacity purification of hen egg-white proteins by ion-exchange electrochromatography with an oscillatory transverse electric field. AB - The ion-exchange electrochromatography with an oscillatory electric field perpendicular to the mobile-phase flow driven by pressure (pIEEC) was used to separate hen egg-white (HEW) proteins. The results were compared with those of normal ion-exchange chromatography (IEC). The column was designed as three compartment rectangular column of 2-mL with dimensions (length x width x depth) of 40 x 10 x 5 mm(3) and the electric field was applied across the direction of column width. Q Sepharose FF was packed into the central compartment as the chromatographic bed. It was confirmed that the dynamic binding capacity (DBC) of different proteins (ovotransferrin and ovalbumin) in the HEW solution increased 2.3 times when an oscillatory electric current of 30 mA at 1/20 Hz was applied in the transverse column direction. Then, the HEW proteins were separated by the pIEEC at loading amounts 2.3-fold higher than those by the IEC. When the feedstock of about one-third of the DBC was applied to the columns (i.e., 7 mL for the pIEEC and 3 mL for the IEC), similar separation efficiencies of the two chromatographic modes were achieved. Both the recovery yield and purity reached 73% to over 90%. The results indicate that the pIEEC is promising for high capacity purification of proteins. PMID- 17120824 TI - Fabrication of PDMS/glass microchips by twofold replication of PDMS and its application in genetic analysis. AB - In this paper, we describe a simple method for fabrication of high quality poly(dimethylsiloxane) (PDMS)/glass microchip by twofold replica molding of PDMS. This technique first served to transfer the negative microchannels from the glass template to the PDMS substrate as a master, and then this PDMS master with positive microchannels was used to replicate the PDMS replica with negative microchannels. Finally, the PDMS replica was bound to a glass sheet by UV radiation. The fabricated microchips were successfully applied for the detection of C677T mutation from the human methylenetetrahydrofolate reductase gene. PMID- 17120825 TI - Separation and quantitation of fructose-6-phosphate and fructose-1 ,6-diphosphate by LC-ESI-MS for the evaluation of fructose-1,6-biphosphatase activity. AB - An LC-ESI-MS method was developed for the identification and quantification of fructose-1,6-biphosphate (F1,6BP) and fructose-6-phosphate (F6P), respectively the substrate and the product of the enzymatic reaction catalysed by fructose-1,6 bisphosphatase (F1,6BPase). F1,6BPase, expressed predominantly in liver and kidney, is one of the rate-limiting enzymes of hepatic gluconeogenesis and has become a target for the development of new drugs for type 2 diabetes. The two sugar phosphates were separated on a Phenomenex Luna NH2 column (150 mm x 2.0 mm id) using the following mobile phase: 5 mM triethylamine acetate buffer/ACN (80:20) v/v in a linear pH gradient (from pH = 9 to 10 in 15 min) at the flow rate of 0.3 mL/min. The detection was performed with an IT mass spectrometer in negative polarity (full scan 100-450 m/z) and in SIM mode on the generated anions at m/z = 339 (F1,6BP) and m/z = 259 (F6P). Under the optimised final conditions, the method was validated for accuracy, specificity, precision (inter- and intradays RSD comprised between 1.0 and 6.3% over the range of concentrations used), linearity (50-400 microM), LODs (0.44 microM) and LOQs (1.47 microM), and the method was applied to F6P determination in the F1,6BPase catalysed hydrolysis of F1,6BP. PMID- 17120826 TI - Column experiments to assess the effects of electron donors on the efficiency of in situ precipitation of Zn, Cd, Co and Ni in contaminated groundwater applying the biological sulfate removal technology. AB - BACKGROUND, AIMS AND SCOPE: In a previous study, we explored the use of acetate, lactate, molasses, Hydrogen Release Compound (HRC, which is based on a biodegradable poly-lactate ester), methanol and ethanol as carbon source and electron donor to promote bacterial sulfate reduction in batch experiments, this with regards to applying an in situ metal precipitation (ISMP) process as a remediation tool to treat heavy metal contaminated groundwater at the site of a nonferrous metal work company. Based on the results of these batch tests, column experiments were conducted with lactate, molasses and HRCI as the next step in our preliminary study for a go-no go decision for dimensioning an on site application of the ISMP process that applies the activity of the endogenous population of sulfate-reducing bacteria (SRB). Special attention was given to the sustainability of the metal precipitation process under circumstances of changing chemical oxygen demand (COD) to [SO4(2-)] ratios or disrupted substrate supply. METHODS: To optimize the ISMP process, an insight is needed in the composition and activity of the indigenous SRB community, as well as information on the way its composition and activity are affected by process conditions such as the added type of C-source/ electron donor, or the presence of other prokaryotes (e.g. fermenting bacteria, methane producing Archaea, acetogens). Therefore, the biological sulfate reduction process in the column experiments was evaluated by combining classical analytical methods [measuring heavy metal concentration, SO4(2-)-concentration, pH, dissolved organic carbon (DOC)] with molecular methods [denaturing gradient gel electrophoresis (DGGE) fingerprinting and phylogenetic sequence analysis] based on either the 16S rRNA-gene or the dsr (dissimilatory sulfite reductase) gene, the latter being a specific biomarker for SRB. RESULTS AND DISCUSSION: All carbon sources tested promoted SRB activity, which resulted within 8 weeks in a drastic reduction of the sulfate and heavy metal contents in the column effluents. However, unexpected temporal decreases in the efficiency of the ISMP process, accompanied by the release of precipitated metals, were observed for most conditions tested. The most dramatic observation of the failing ISMP process was observed within 12 weeks for the molasses amended column. Subsequent lowering the COD/ SO4(2-) ratio from 1.9 to 0.4 did not alter the outcome of sulfate reduction and metal precipitation efficiency in this set-up. Remarkably, after 6 months of inactivity, bacterial sulfate reduction was recovered in the molasses set up when the original COD/ SO4(2-) ratio of 1.9 was applied again. Intentional disruption of the lactate and HRC supplies resulted in an immediate stagnation of the ISMP processes and in a rapid release of precipitated metals into the column effluents. However, the ISMP process could be restored after substrate amendment. 16S rDNA-based DGGE analysis revealed that the SRB population, in accordance with the results of the previously performed batch experiments, consisted exclusively of members of the genus Desulfosporosinus. The community of Archaea was characterized by sequencing amplicons of archaeal and methanogen-specific PCR reactions. This approach only revealed the presence of non-thermophilic Crenarchaeota, a novel group of organisms which is only distantly related to methane producing Euryarchaeota. DGGE on the dsrB genes was successfully used to link the results of the ISMP process to the community composition of the sulfate reducing bacteria. CONCLUSIONS: In the case of an intentional disruption of substrate supply, the ISMP process failed most likely because the growth and activity of the indigenous SRB community stopped due to a lack of a carbon and electron donor. On the other hand, the cause of the sudden temporal shortcomings of the ISMP process in the presence of different substrates was not immediately clear. It was first thought to be the result of competition between methanogenic prokaryotes (MP) and sulfate reducers, since the formation of small amounts of CH4 (0.01-0.03 ppm ml(-1) was detected. However, the results of molecular analyzes indicate that methanogens do not constitute a major fraction of the microbial communities that were enriched in the column experiments. Therefore, we postulate that the SRB population becomes inhibited by the formed metal sulfides. RECOMMENDATION AND PERSPECTIVE: Our results indicate that the ISMP process is highly dependent on SRB-stimulation by substrate amendments and suggest that this remedial approach might not be viable for long-term application unless substrate amendments are continued and environmental conditions are strictly controlled. This will include the removal of affected aquifer material from the metal precipitation zone at the end of the remediation process, or removal of metal precipitates when the microbial activity decreases. Additional tests are necessary to investigate what will happen when clear groundwater passes through the reactive zone while no more C-sources are amended and all indigenous carbon is consumed. Also, the effects of dramatic increases in sulfate- or HM-concentrations on the SRB-community and the concomitant ISMP process need to be studied in more detail. PMID- 17120827 TI - Characterization of primary precipitate composition formed during co-removal of Cr(VI) with Cu(II) in synthetic wastewater. AB - BACKGROUND, AIMS AND SCOPE: Hexavalent chromium [Cr(VI)] cannot react with either carbonate or hydroxide to form chromium precipitates. However, by using a precipitation technology to treat plating wastewater containing Cr(VI), Cu(II), Ni(II) and Zn(II), approximately 78% of Cr(VI) (initial 60 mg/L) was co-removed with the precipitation of Cu(II), Ni(II) and Zn(II) (each 150 mg/L) by dosing with Na2CO3 (Sun 2003). Direct precipitation by forming Cu(II)-Cr(VI) precipitates followed by adsorption of Cr(VI) onto freshly formed Cu-precipitates was subsequently found to be the main mechanism(s) involved in Cr(VI) co-removal with Cu(II) precipitation by dosing Na2CO3 stepwise to various pH values (Sun et al. 2003). This study was. carried out to further characterize the formation of primary precipitates during the early stages of copper precipitation and simultaneous removal of Cr(VI) with Cu(II). METHODS: Test metal-solutions were prepared with industrial grade chemicals: CuCl2 x 2H2O, Na2SO4 and K2Cr2207. NaCO3 was added drop-wise to synthetic metal-solution to progressively increase pH. For each pH increment, removal of soluble metals was detected by atomic absorption spectrophotometer (AAS) and surface morphology of precipitates was analyzed by scanning electron microscope (SEM). To further characterize the formation of primary precipitates, a series of MINEQL+ thermodynamic calculations/analyses and equilibrium calculations/ analyses were conducted. RESULTS AND DISCUSSION: MINEQL+ thermodynamic calculation indicated that, for a system containing 150 mg/L Cu(II) and 60 mg/L Cr(VI) with gradual Na2CO3 dosing, if any precipitates can be formed at pH 5.0 or lower, it should be in the form of CuCrO4. Comparison tests using systems containing the same equivalent of Cu(II) plus Cr(VI) and Cu(II) plus SO4(2-) showed that the precipitation occurred at a pH of around 5.0 in the Cu(II)-Cr(VI) system and around 6.0 in the Cu(II)-SO4(2-) system. The discrepancy of the precipitation was indeed caused by the formation of Cu-Cr precipitates. The initiation of copper removal at pH around 5.0 for the Cu-Cr co-removal test was not attributable to the formation of Cu-CO3 precipitates, instead, it was most likely through the formation of insoluble Cu Cr precipitates, such as CuCrO4 and CuCrO4 x 2Cu(OH)2. Experimental tests, equilibrium calculations, MINEQL+ thermodynamic calculations and surface morphologies for systems using higher concentrations of Cu(II) and Cr(VI) further verified the most probable composition of primary precipitates is copper chromate. CONCLUSION: In the Cu-Cr co-removal test with Na2O3 dosing to increase pH and induce metal precipitation, copper-chromate precipitates are the primary precipitates produced and contribute to the initial simultaneous removal of copper and chromium. PMID- 17120828 TI - Leaching potential of some phenylureas and their main metabolites through laboratory studies. AB - BACKGROUND, AIMS AND SCOPE: Laboratory studies were conducted with the aim of defining the leaching potential of some phenylureas and their metabolites. A first study was performed for calculating their leaching index (as GUS) on the base of intrinsic properties: persistence (as DT50) and mobility (as Koc) in soil. Another study consisted of aged column leaching experiments whose meaning was to semi-quantify the occurrence of the tested compounds in the leachates, so simulating in field conditions. METHODS: The tested compounds were: diuron, linuron and monolinuron (parents); 3,4-dichloroaniline (DCA), 4-chloroaniline (CLA), 1-(3,4-dichlorophenyl)-3-methylurea (DCPMU), 1-(3,4-dichlorophenyl)urea (DCPU), 1-(4-chlorophenyl)urea (CPU) and monuron, this latter considered both as a metabolite and parent compound. The Koc values of the examined substances were determined by the HPLC screening methods, according to the OECD TG 121. DT50 determinations and aged column leaching experiments were carried out according to SETAC procedures. RESULTS AND DISCUSSION: The examined compounds showed a rather wide range of persistence in soil, with DT50 values less than 2 days for DCA and CLA, close to 8 days for DCPU and CPU and from 16 (diuron) up to 24.8 (DCPMU) days for the others. Their mobility was generally high, based on their Koc values, which ranged from 33 (CPU) to 406 (linuron). The GUS indices indicated that monuron has a clear potential to contaminate groundwater (> 2.8); DCPMU, monolinuron, CPU and diuron are intermediate contaminants (1.8-2.8). Linuron, DCPU, CLA and DCA exhibited a non-leaching behaviour (< 1.8). The aged leaching column experiments showed that parents were found in the leachates at very high percentages respect to the doses applied. The metabolites reached much less percentages, the highest values were observed for monuron from diuron (5.7), CPU (7.2) and DCPMU (8.2%). CONCLUSION: Diuron, Monuron, CPU and DCPMU on the basis of their intrinsic properties, formation from their parents and occurrence in leachates from aged column leaching studies, seem to possess the characteristics of groundwater contaminants. The methodological approach of this study is relatively easy and rapid, hence it can represent a tool for a first screening of compounds such as pesticide metabolites (generally available only in small quantities and for which a field study is not conceivable) or other compounds for which not adequate environmental data are available. PMID- 17120829 TI - In-situ surfactant/surfactant-nutrient mix-enhanced bioremediation of NAPL (fuel) contaminated sandy soil aquifers. AB - SCOPE AND BACKGROUND: Contamination of soils, aquifers and groundwater by nonaqueous phase liquid (NAPL) pollutants constitutes a major environmental issue of concern, worldwide. The residual (biodegradation-resistant) hydrophobic fuel hydrocarbons entrapped in the soil porous matrix, possess a particular bioremediation challenge due to their becoming virtually immobile, nor desorbable, or water dispersible. Consequently, they are not available as substrates to the micro-organism-based biodegradation. MATERIALS AND METHODS: Our research involves the development of economically feasible, surfactant/surfactant nutrient mix (SSNM)-enhanced bioremediation methodologies for sustainable, in situ bioremediation of fuel-contaminated aquifers. This requires, methodologically, (a) the optimization, via in vitro 'flow' (columns) lab experiments and screening processes, of an effective mixture for the intended SSNM-enhanced bioremediation; and (b) the study of the combined effect of the optimized SSNM on the solubilization/mobilization and biodegradation of NAPL (fuel) in in vitro site/aquifer-simulated bioremediation. RESULTS AND DISCUSSION: The essence of our findings: (1) kerosene's maximum enhanced mobilization - f = 3.6, compared with that of deionized water, was achieved with an SSNM having the composition of linear alkylbenzene sulfonate (LABS): coco-amphodiacetate (containing N): surfactant-nutrient X (containing both N and P) = 0.15: 0.15: 0.05 g/L, respectively; (2) 62-64% of the initial amount of kerosene in the initially saturated soil matrix, 'packed' in a column, has been eluted from it during approximately 30 days, compared with 68% of kerosene biodegradation in 'vessel' settings, in 21 days. CONCLUSIONS: (1) The indigenous microorganisms present in th vadose zones of fuel-contaminated sandy soil aquifers are potentially capable of unassisted removal of approximately 80% of the initially contained fuel (kerosene), during a period of about 42 days; (2) the major effects of the SSNM addition are (a) enhanced mobilization of the bulky NAPL; and (b) enhanced desorbtion/ solubilization/dispersion of the entrapped NAPL which, in turn, facilitate their enhanced biodegradation. RECOMMENDATIONS AND PERSPECTIVE: Our findings suggest that pre-optimized, biodegradable SSNM is essential for surfactants-based bioremediation of NAPL-contaminated aquifers, in order to make this in-situ methodology both technologically and economically feasible. PMID- 17120830 TI - Pilot study on road traffic emissions (PAHs, heavy metals) measured by using mosses in a tunnel experiment in Vienna, Austria. AB - BACKGROUND, AIMS AND SCOPE: Over the last few years there has been extensive research for new indicators providing information about deposition resulting from road traffic and tunnel experiments received special attention in emission research. Mosses have been used for the estimation of atmospheric heavy metal and PAH depositions for more than three decades, although they were used only a few times for estimating ambient air pollution caused by traffic. In the current study, the suitability of using a moss species for monitoring road traffic emissions inside a tunnel was evaluated. This was a first-time ever attempt to use plants (mosses) as bioindicators in a tunnel experiment. Specifically, two relevant questions were examined: 1) Do mosses accumulate toxic substances derived from road traffic emissions under the extremely adverse conditions which can be found in a tunnel, and 2) Which substances can mainly be attributed to road traffic emissions and therefore be taken as efficient and reliable indicators for motor vehicles? METHODS: For the first time a biomonitor (the moss species Hylocomium splendens (Hedwig) B.S.G.) was used in a road tunnel experiment to analyse emissions from road traffic. Moss samples were exposed for four weeks in wooden frames (size 10 cm x 10 cm), covered by a thin plastic net with a mesh size of 1 cm x 1 cm. 17 elements, mainly heavy metals, and the 16 EPA PAHs together with coronene were analysed by ICP-AES, AAS and GC-MSD. RESULTS: Enrichment factors, calculated by comparing post-experiment concentrations to those of a background site, were high for most PAHs, especially benzo(g,h,i)perylene (150.7), coronene (134.7), benzo(a)anthracene (125.0), indeno(1,2,3-c,d)pyrene (79.8), chrysene (78.1), pyrene (69.6) and benzo(b) fluoranthene (67.4), and among the other elements for Sb (73.1), Mo (59.6), Cr (33.9), As (24.1), Cu (19.6), and Zn (17.1). All these substances can thus be taken as indicators for road traffic pollution. Concentrations were also significantly higher in the tunnel mosses for all investigated substances than along busy roads outside tunnels. Cluster analysis revealed groups of substances which could sensibly be attributed to various sources (abrasion processes, Diesel combustion) and enrichment in the various particle size classes. DISCUSSION: The extreme high concentrations in the analysed moss samples from inside the tunnel were due to higher concentrations in the ambient tunnel air, and the fact that already deposited chemical substances are not lost by rain, as well as efficient uptake capacities even under the extremely adverse conditions in a tunnel. In accordance with previous studies our results suggest that PAHs are better indicators for emissions from the burning process than heavy metals. CONCLUSIONS: As in open fields, mosses are suitable indicators for monitoring traffic emissions in tunnels. In addition to biomonitoring in open fields, in tunnel experiments mosses are even better indicators, because the confounding effects of other sources of pollution and the 'noise' in the accumulation process (e.g. washout through wet deposition) are minimised. The results of our study demonstrate the usefulness of mosses for surveying heavy metals and PAH emissions and deposition arising from road traffic sources, even under the extremely adverse conditions of the tunnel environment. RECOMMENDATION: It can be considered that biomonitors like mosses are a suitable alternative to technical particle filters inside tunnels. They are easy to handle, low in costs and valuable information regarding traffic emissions can be obtained. PERSPECTIVE: The results of this pilot-study proved the feasibility of the method, however, should be corroborated by further investigations based on a sample set that allows for generalization of the findings and might even include other moss species. A comparison of technical measurements with the biomonitoring method could lead to a more general acceptance of the results. PMID- 17120831 TI - The origin of stable halogenated compounds in volcanic gases. AB - BACKGROUND: Halogenated compounds in the atmosphere are of great environmental concern due to their demonstrated negative effect on atmospheric chemistry and climate. Detailed knowledge of the emission budgets of halogenated compounds has to be gained to understand better their specific impact on ozone chemistry and the climate. Such data are also highly relevant to guide policy decisions in connexion with international agreements about protection of the ozone layer. In selected cases, the relevance of specific emission sources for certain compounds were unclear. In this study we present new and comprehensive evidence regarding the existence and relevance of a volcanic contribution of chlorofluorocarbons (CFCs), hydrofluorocarbons (HFCs), hydrochlorofluorocarbons (HCFCs), halons (bromine containing halo(hydro)carbons), and fully fluorinated compounds (e.g. CF4 and SF6) to the atmospheric budget. METHODS: In order to obtain new evidence of a volcanic origin of these compounds, we collected repeatedly, during four field campaigns covering a period of two years, gases from fumaroles discharging over a wide range of temperatures at the Nicaraguan subduction zone volcanoes Momotombo, Cerro Negro and Mombacho, and analysed them with very sensitive GC/MS systems. RESULTS AND DISCUSSION: In most fumarolic samples certain CFCs, HFCs, HCFCs, halons, and the fully fluorinated compounds CF4 and SF6 were present above detection limits. However, these compounds occur in the fumarole gases in relative proportions characteristic for ambient air. CONCLUSION: This atmospheric fingerprint can be explained by variable amounts of air entering the porous volcanic edifices and successively being incorporated into the fumarolic gas discharges. Recommendation and Outlook. Our results suggest that the investigated volcanoes do not constitute a significant natural source for CFCs, HFCs, HCFCs, halons, CF4, SF6 and NF3. PMID- 17120832 TI - Effects of zeolites on cultures of marine micro-algae: A brief review. AB - GOAL, SCOPE AND BACKGROUND: The cation-exchange capacity of zeolites is well known and has been increasingly explored in different fields with both economic and environmental successes. In aquatic medium with low salinity, zeolites have found multiple applications. However, a review of the literature on the applications of zeolites in salt waters found relatively few articles, including some recently published papers. The purpose of this review is to present the state-of-the-art on applications of using zeolites for amending the trace elemental contents of salt water as well as the implications of this property for promoting marine micro-algal growth. MAIN FEATURES: This paper deals with the following features: Sorption capacity of zeolites including 1. application of zeolites in saltwater, 2. the role of silicon and zeolites on cultures of micro algae, and 3. the role of organically chelated trace metals. RESULTS: The following competing factors have been identified as effects of zeolites on algal growth in salt water: (i) ammonia decrease: growth inhibition reduced; (ii) macro nutrients increase, mainly silicon: stimulation of silicon-dependent algae; (iii) trace metals increase (desorption from zeolites) or decrease (adsorption): inhibition or stimulation, depending on the nature of the element and its concentration; and, (iv) changes in the chelating organics exudation: inhibition or stimulation of growth, depending on the (a) nature of the complexed element; (b) bioavailability of the complex; and (c) concentration of the elements simultaneously present in inorganic forms. DISCUSSION: Zeolites have been capable of stimulating the growth of the silicon-demanding marine micro-algae, like diatoms, mainly because they can act as a silicon buffer in seawater. Zeolites can also influence the yield of non-silicon-demanding algae, because the changes they can cause (liberation and adsorption of trace elements) in the composition of the medium. CONCLUSIONS: Zeolites have been capable of stimulating the growth of the marine micro-algae. However, the extent of ion exchange between zeolite and seawater, which conditions the effects, will depend on several factors: (1) initial metal concentration in seawater; (2) levels of trace metals in the zeolites (contaminants); (3) characteristics of the zeolites in terms of both ion exchange capacity and specific affinities for the different cations; (4) quantity of zeolite per litre of solution; (5) pH and (6) response of the organism in terms of liberation of organic ligands. RECOMMENDATIONS AND PERSPECTIVES: RECOMMENDATIONS: Therefore, a previous investigation in each particular case is recommended, in order to select the zeolitic characteristics and concentrations that will maximize the algal yield. PERSPECTIVES: Stimulation of phytoplankton growth can be economically relevant since phytoplankton constitutes the basis of the marine food webs and is required in fish farming nurseries in the marine aquaculture industry. Zeolites are cheap, only small amounts (few milligrams per liter of culture) are required and the addition of some micro-nutrients may be omitted. Therefore, the inclusion of zeolites in algal cultures in aquaculture may have economic advantages. PMID- 17120833 TI - Range-wide Yangtze freshwater dolphin expedition: The last chance to see Baiji? AB - BACKGROUND, AIM AND SCOPE: There are two species of fresh water cetaceans surviving in the Yangtze River system in China: Baiji (Lipotes vexillifer) and Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis). As a result of the expansion of human activities on the river, their distribution ranges appear to be decreasing and in the case of the Baiji, are even being restricted to several sections. The Baiji is the world's most critically endangered cetacean species with a population estimated at only a few tens of individuals. The Yangtze finless porpoise is the world's only freshwater-adapted population of the species, and it has been estimated that only around 1,000 individuals remain in the river system. In order to prevent the extinction of Baiji and a sharp decline in the abundance of the porpoise, in situ conservation (i.e. in the river) and two ex situ conservation (i.e. in semi-natural reserves and in captivity) strategies were proposed and have been implemented since the early 1990s. In view of both the severely endangered status of the animals and the severely degraded conditions of their habitats, the feasibility and actual status of these two strategies are raised for discussion. MAIN FEATURES: The threats faced by the cetaceans are mainly from the unfettered exploitation of the river's resources. In the past 20 years, five nature reserves have been established along the river. Imposing maximum prohibition of harmful and illegal fishing methods in the reserves might prolong the process of extinction of these cetaceans in the wild, but so far, the administrative measures taken in the reserves have not yet kept the abundance from sharply declining. As human use of the river and its resources is expected to intensify for many decades into the future, the ability of the river to continue to support these species is certainly undecided. Therefore, rescuing animals from the river and establishing viable breeding populations in seminatural reserves, in which the environment is similar to the main stream of the river, and in captivity, has to be considered urgently as the short-term goal of ex situ strategies. Since the abundance of porpoises is higher than that of the Baiji, we have first established breeding populations of them in the semi natural reserves and in captivity. But, considering the extremely low density of Baiji in the river, an immediate range-wide Yangtze Baiji survey is an urgent need for locating and capturing sufficient Baiji for successfully establishing a breeding population of them in semi-natural reserves. RESULTS: Two semi-natural reserves (in Shishou, Hubei Province, and Tongling, Anhui Province) have been set up along the river in order to establish breeding populations of the Baiji and the porpoises. So far, several small groups of porpoises that were caught in the main stream of the river have successively been introduced into the semi-natural reserves. Under careful management, these animals in both of the semi-natural reserves not only survive, but can also reproduce naturally and successfully. At least one or three calves were born in each reserve each year. Additionally, a breeding group of porpoises is being established at the Baiji Dolphinarium at the Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan. There are presently four adults and one calf living in the Dolphinarium. The calf, born in July, 2005, is the first captive bred Yangtze Finless Porpoise in the world. In preparation for the range-wide Yangtze cetacean survey, a 9-day pilot expedition on the river near Wuhan was conducted in March, 2006, in order to develop methods for locating the Baiji. No Baiji were expected to be seen in such a short period but about 40 porpoise sightings were observed. Results of the pilot survey indicated that traditional visual and acoustical survey methods for cetaceans should be adapted to find the elusive Baiji in the river. Currently, the range wide Yangtze cetacean survey is in preparation. The survey will cover over 1,700 km of the Yangtze River from Yichang to Shanghai, and is expected to provide detailed information on Baiji and porpoise numbers and distribution patterns in the river. DISCUSSION: Although the short-term goal of ex situ conservation is to rescue cetaceans from the river and to establish viable breeding populations in semi-natural reserves and in captivity, the long-term goal of releasing the animals back into the river when the threats have decreased and the natural environment has been improved, should not be neglected. Moreover, the in situ conservation efforts in the natural reserves, and even in the entire Yangtze River system, including the lakes, should not be ignored or abandoned at any time. The activities contributing to the conservation of the Baiji and the porpoise in the wild have the incidental effect of benefiting the entire Yangtze ecosystem and other rare threatened species. The dynamics of the groups of porpoises in semi-natural reserves should be monitored continually, in order to guide the establishment of breeding groups of Baiji in these semi-natural reserves in the near future. CONCLUSIONS: Under the existing severely degraded conditions of the Yangtze system, the sharply fall populations of Baiji and porpoises will not be suspended in the foreseeable future. Therefore, ex situ conservation should be emphasized, and the severely threatened Baiji in the river should be removed and translocated to semi-natural reserves for establishing viable breeding populations. The successful program of capturing, translocating and maintaining finless porpoises in the Shishou semi-natural reserve has demonstrated its adequacy as an ex situ environment for cetaceans. Following the successful pilot survey in the river, the immediate range-wide Yangtze cetacean survey is proposed and is in preparation. The range-wide survey is expected to ensure that any remaining Baiji can be found reliably and captured successfully after the survey. RECOMMENDATIONS AND PERSPECTIVES: During the range-wide survey, not only the Baiji but also the porpoise as well as their habitats should be investigated based on visual and acoustical methods that adapted to the river and the animals. Meanwhile, the current risk levels to the Baiji and porpoises should be evaluated at each area where Baiji or porpoises can be reliably sighted. Any capture efforts should be targeted on the most threatened areas, or where there is maximum risk of injury or death. The immediate track of the Baiji should be carried out once a Baiji is sighted during the range-wide survey in order to obtain the movement route of the animals, which is crucial information for the successful capture operation. Additionally, the need to establish new semi natural reserves for the porpoises should be placed on the agenda of local and central governments in the near future. PMID- 17120834 TI - Effect thresholds and 'adequate control' of risks: The fatal flaws in the EU council's position on authorisation within REACH. AB - BACKGROUND: Preparation of the new European REACH (Registration, Evaluation and Authorisation of CHemicals) regulation on chemicals has reached a critical stage. Depending on how key elements of the legislative proposal are finalised, especially that on authorisation of uses of 'substances of very high concern', REACH could either provide an effective measure to drive innovation towards cleaner and safer alternatives, or instead lead to further avoidable chemical exposures on the basis of demonstrated 'adequate control' of risks. Given that some key indicators of human and wildlife reproductive health continue to decline in parts of Europe, while evidence for chemical exposure as a contributory factor grows, it will clearly be vital to get the legislation right. GOAL AND SCOPE: Whereas there is now a consensus between the European Parliament and Council of the European Union that uses of persistent, bioaccumulative and toxic (PBT) and very persistent and very bioaccumulative (vPvB) substances should only be permitted when no safer alternatives are available, major differences remain regarding the manner in which other 'substances of very high concern' (including substances which are carcinogenic, mutagenic or toxic to reproduction (CMRs) and endocrine disruptors) are addressed. This paper examines those differences in more detail and proposes some ways forward. METHODS: Using case studies of specific chemicals as examples, the paper critically evaluates the concepts of 'effect thresholds' and 'adequate control' of risks, which underpin the Council's proposal for many CMRs and endocrine disruptors. RESULTS: The subjectivity and uncertainties inherent in the threshold approach proposed by the Council, as illustrated by these case examples, bring its ability to ensure a high level of protection for the environment and human health into question: i. the nature and extent of toxic effects recorded depend on many different factors, including the type of test and conditions selected, the organisms exposed, the timing of exposure and precisely which effects are measured and over what timeframe. ii. doses considerably below 'no effect levels' for survival could nevertheless be causing significant impairment to health and/or reproductive success. iii. chemicals present in mixtures at levels below established thresholds for effects may, in combination, induce significant toxicological responses. DISCUSSION: Under the Council's current proposal, companies will be granted authorisations for some uses of CMRs and endocrine disruptors, even if safer alternatives without these properties are already on the market. The high level of evidence required for identification of substances as being of equivalent concern represents an additional weakness in the Council approach. CONCLUSIONS: Instead, a requirement (along the lines of the Parliament's proposals) to address the availability of alternatives in all cases, to use them when available and to initiate their development when not, represents a more robust, defensible and protective approach to the management of 'substances of very high concern'. The possibility for authorisation of essential uses would remain, while all avoidable uses and exposures would progressively be prevented and sustainable innovation supported. PERSPECTIVES: In the long run, this can only lead to a more sustainable future for the chemical industry in Europe, as well as delivering benefits of increased protection for our environment and health for generations to come. PMID- 17120835 TI - Initiative for an International Panel on Chemical Pollution (IPCP). PMID- 17120836 TI - Experimental studies of environmental processes: A practical course in environmental chemistry. AB - BACKGROUND, AIM AND SCOPE: To predict the fate of persistent organic pollutants in the environment, several aspects have to be considered carefully: their basic physical and chemical properties, distribution, transport within and among compartments, biotic and abiotic transformation processes, as well as effects on living organisms including humans. Laboratory simulation of the processes that control the chemical behavior of organic compounds in the environment is often desirable for deeper understanding, as well as for determination of basic characteristics required for successful environmental modeling. However, laboratory exercises targeted on the simulations of environmental processes are very rare. MATERIALS AND METHODS: Practical training was designed as a supplement to the course of Environmental Chemistry. The whole course consists of seven experiments in a logical sequence-partition coefficient (n-octanol-water), Henry's Law Constant, soil sorption, volatilization from soil, bioaccumulation, photochemical degradation and microbial degradation. The objective is to gain knowledge of the principal physical processes, by which chemicals move, concentrate and dissipate, as well as of the principal routes of biotic and abiotic degradation. RESULTS: New practical course reflecting current topics and approaches of environmental chemistry was introduced to M.S. students at Masaryk University in 2004. Detailed description of the course including experimental techniques is provided in this article. DISCUSSION: Lab course employs basic techniques for the study of environmental processes to strengthen links between the theory taught in the lectures of Environmental Chemistry, laboratory simulations, and processes in the real word. CONCLUSIONS: A significant effort towards developing this laboratory exercise was devoted with the goal of providing a context for teaching both fundamental theories and environmental techniques, strengthening the link between the theory and processes in the real world, providing a scientific background for understanding the environmental problems, challenging the students with the experience of simulating the physical, chemical, and biological processes that control contaminant fate and transport in the environment, showing the students importance of the interdisciplinary approach and giving the students a taste of the excitement and challenge of the research in real conditions. RECOMMENDATIONS AND PERSPECTIVES: The laboratory course was taught with a great success, and it became a part of recommended study plans for M.S. students in Environmental Chemistry and Ecotoxicology. Suggested experiments proved to be suitable for simulation of biotic and abiotic transformations, bioaccumulation, as well as distribution and transport processes in a student laboratory with good reproducibility and accuracy. The response of the students was very positive. PMID- 17120837 TI - Artful forgery. PMID- 17120838 TI - The time course of visual afterimages: data and theory. AB - Sequential viewing of two orthogonally related patterns produces an afterimage of the first pattern (Vidyasagar et al, 1999 Nature 399 422-423; Francis and Rothmayer, 2003 Perception and Psychophysics 65 508-522). We investigated how the timing between the first stimulus (a vertical bar grating) and the second stimulus (a horizontal bar grating) affected the visibility of the afterimage (a perceived vertical grating). As the duration from offset of the first stimulus increased, reports of afterimages decreased. Holding fixed the total time from offset of the first stimulus and increasing the duration from offset of the second stimulus while decreasing the time between the first and second stimuli, caused a decrease in afterimage reports. We interpret this finding in terms of Grossberg's BCS - FCS (boundary contour system--feature contour system) theory. In this theory, the afterimage percept is the result of color complement after responses in the FCS system interacting with orientation after-responses in the BCS system. The two types of after-responses interact at a stage of neural filling-in to produce the afterimage percept. As the duration between the stimuli increases, the color after-responses weaken so that visible filling-in is less likely to occur. A similar effect occurs for the orientation after-responses but at a faster time scale. Simulations of the model match the experimental data. PMID- 17120839 TI - The influence of depth segmentation on colour constancy. AB - In real scenes, surfaces in different depth planes often differ in the luminance and chromatic content of their illumination. Scene segmentation is therefore an important issue when considering the compensation of illumination changes in our visual perception (lightness and colour constancy). Chromatic adaptation is an important sensory component of colour constancy and has been shown to be linked to the two-dimensional spatial structure of a scene (Werner, 2003 Vision Research 43 1611 - 1623). Here, the question is posed whether this cooperation also extends to the organisation of a scene in depth. The influence of depth on colour constancy was tested by introducing stereo disparity, whereby the test patch and background were perceived in either the same or one of five different depth planes (1.9-57 min of arc). There were no additional cues to depth such as shadows or specular highlights. For consistent illumination changes, colour constancy was reduced when the test patch and background were separated in depth, indicating a reduction of contextual influences. An interaction was found between the influences of stereo depth and spatial frequency on colour constancy. In the case of an inconsistent illumination change, colour constancy was reduced if the test patch and background were in the same depth plane (2-D condition), but not if they were separated in depth (3-D condition). Furthermore, colour constancy was slightly better in the 3-D inconsistent condition than in the 2-D inconsistent condition. It is concluded that depth segmentation supports colour constancy in scenes with inconsistent illumination changes. Processes of depth segmentation are implemented at an early sensory stage of colour constancy, and they define visual regions within which the effects of illuminant changes are discounted for separately. The results support recent models that posit such implementation of scene segmentation in colour constancy. PMID- 17120840 TI - Lightness of an object under two illumination levels. AB - Anchoring theory (Gilchrist et al, 1999 Psychological Review 106 795-834) predicts a wide range of lightness errors, including failures of constancy in multi-illumination scenes and a long list of well-known lightness illusions seen under homogeneous illumination. Lightness values are computed both locally and globally and then averaged together. Local values are computed within a given region of homogeneous illumination. Thus, for an object that extends through two different illumination levels, anchoring theory produces two values, one for the patch in brighter illumination and one for the patch in dimmer illumination. Observers can give matches for these patches separately, but they can also give a single match for the whole object. Anchoring theory in its current form is unable to predict these object matches. We report eight experiments in which we studied the relationship between patch matches and object matches. The results show that the object match represents a compromise between the match for the patch in the field of highest illumination and the patch in the largest field of illumination. These two principles are parallel to the rules found for anchoring lightness: highest luminance rule and area rule. PMID- 17120841 TI - Determinants of perceived surface transparency in two-dimensional achromatic textured patterns. AB - Achromatic transparency in 2-D surfaces composed of three adjacent areas, one created from the others, occurs when in the created area it is possible to see the two colours of the adjacent areas. Displays with two white and black intersecting bars were produced to verify the possibility of perceiving transparency in the intersection area when this was subdivided into a different number of white and black diamonds. By increasing the spatial frequency of the elements in the intersection area we investigated the impression of transparency, and found that (i) the impression of transparency increases up to a spatial frequency of 10 cycles deg(-1) and then levels off, (ii) there is a transition from a perception of transparency in which white and black are well discriminable to a perception in which a perceptually uniform grey is seen, (iii) the perception of transparency occurring when colours are discriminated seems consistent with Helson's area-luminance hypothesis [1964, in Adaptation Level Theory (New York: Harper & Row) pp 282-292], according to which the increase of spatial frequency determines the transition from the phenomenon of contrast to that of assimilation. PMID- 17120842 TI - Stereo motion transparency processing implements an ecological smoothness constraint. AB - Transparent motion stimuli allow us to investigate how visual motion is processed in the presence of multiple sources of information. We used stereo random-dot kinematograms to determine how motion processing is affected by the difference in direction and depth of two overlapping motion components. Observers judged whether a noise dot display contained one or two directions of motion. For all disparity differences, performance did not change among angles greater than 60 degrees, but the ability to detect transparent motion fell dramatically as the direction difference decreased below 60 degrees. When a disparity difference was added between the two motion components, detection became easier. We compared these results to an ideal-observer model limited by stimulus uncertainty and low level sources of internal noise. The resulting measure of efficiency--the ratio of human to model performance--reflects changes in how motion stimuli are being processed. A decrease of both the direction and disparity differences had the effect of decreasing efficiency. These results suggest that the mechanism processing transparent motion may implement a smoothness constraint that tends to combine similar motions into a single percept. PMID- 17120843 TI - Alignment effect: primary-secondary learning and cognitive styles. AB - The degree to which the way of learning spatial information (primary/secondary learning) and spatial cognitive style (landmark/route/survey) affect orientation specificity (alignment effect) is studied. We think that the most important factor explaining the absence of the alignment effect is the spatial cognitive style. We hypothesise that while landmark participants show an alignment effect after both primary and secondary learning, route participants show this effect only after secondary learning, and survey participants do not show it at all. Participants performed three tasks in order to distinguish their cognitive style; they were then randomly assigned to primary or secondary learning and submitted to directional judgment tasks to verify whether the alignment effect was present. The results confirm our hypothesis. PMID- 17120844 TI - Development of modal and amodal completion in infants. AB - Visual completion has been divided into two types: modal and amodal. While psychophysical studies with adults provided several common properties between modal and amodal completion, studies with infants showed differential trends in the development of these perceptual abilities. In the present study, we further examined the development of these two kinds of visual completion in infants aged 3 to 6 months. We created a display composed of a partially overlapping circle and square. The display induced either modal or amodal completion depending on the colour. Infants were familiarised with either the modal or the amodal display. After this familiarisation, the infants were tested on their discrimination between the complete figure and the broken figure. If the infants could perceptually complete the figures in the familiarisation display, they were expected to show a novelty preference for the broken figure. A total of thirty two infants participated in the present study. Our results suggest that modal completion develops by 3-4 months of age, whereas amodal completion develops by 5 6 months of age. PMID- 17120845 TI - Errors in judging information about reflections in mirrors. AB - We investigated people's perception and knowledge of planar mirror reflections. People were accurate at deciding when they could first see their reflection as they approached a mirror from the side, but only if their reflection was visible. Most people stopped too early if the mirror was covered up. People also overestimated the size of the reflection of their face on the surface of a mirror if they were shown a covered mirror. Their accuracy improved somewhat if their reflection was visible but, unlike the first task, they still made striking errors. Perceptual feedback thus improved performance at predicting the behaviour of mirror reflections in both tasks but failed to eliminate errors in the second task. The overestimation of reflection size was not face-specific as it generalised to novel stimuli (paper ellipses) and it was found with both a matching response and for verbal size estimations. The early error in the first task appears to be due to an inaccurate belief that can be overridden by perceptual feedback. The overestimation in the second task is primarily caused by a powerful size-constancy effect. PMID- 17120846 TI - Presentation of a visual nearby moving object alters stream/bounce event perception. AB - Two identical visual objects moving across each other in a two-dimensional display can be perceived as either streaming through or bouncing off each other. The bouncing event percept is promoted by the presentation of a brief sound at the point of coincidence of the two objects. In this study, we examined the effect of the presence of a moving object near the two objects as well as the brief sound on the stream/bounce event perception. When both the nearby moving object and brief sound were presented, a streaming event, not a bouncing event, was robustly perceived (experiment 1). The percentage of the streaming percept was also systematically affected by the proximity of the nearby object (experiment 2). These results suggest that the processing of intramodal grouping between a nearby moving object and either of the two objects in the stream/bounce display interferes with crossmodal (audiovisual) processing. Moreover, we demonstrated that, depending on the trajectory of the nearby moving object, the processing of intramodal grouping can promote the bouncing percept, just as crossmodal processing does (experiment 3). PMID- 17120847 TI - Nursing history: blurring disciplinary boundaries. PMID- 17120848 TI - Rewriting nursing history--again? PMID- 17120849 TI - Florence Nightingale's nursing practice. PMID- 17120850 TI - The nursing radicalism of the honourable Albinia Brodrick, 1861-1955. PMID- 17120851 TI - "The ultimate destination of all nursing": the development of district nursing in England, 1880-1925. PMID- 17120852 TI - "Much instruction needed here": the work of nurses in rural Wisconsin during the Depression. PMID- 17120853 TI - Reweaving a tapestry of care: religion, nursing, and the meaning of hospice, 1945 1978. PMID- 17120854 TI - The rise and demise of the Colonial Nursing Service: British nurses in the colonies, 1896-1966. PMID- 17120855 TI - Florence Nightingale's opposition to state registration of nurses. PMID- 17120856 TI - New tools and interventions take aim at pediatric obesity. PMID- 17120857 TI - Unprecedented region-wide initiative takes on diabetes. AB - Western Pennsylvania gets serious about diabetes with unprecedented effort. It's called the PRIDE initiative, and it has assembled an impressive array of resources to equip primary care practices in the region with the tracking tools they need to bring diabetes care in line with the chronic care model. It's a tall order, and the initiative has just begun, but developers are determined to translate their research into measurable results. PMID- 17120858 TI - The benefits of aggressive BP control in patients with CAD. AB - Is it time to get more aggressive with BP control? Evidence is mounting that the answer to that question is definitely yes. In fact, new research suggests that national guidelines simply don't go far enough with their recommended BP targets for patients who already have cardiac disease. More evidence would be nice, but investigators maintain there is already a compelling case for clinicians to treat to lower targets. PMID- 17120859 TI - New four-pronged approach to wellness combines an eclectic mix of strategies. AB - Humana uses non-traditional means in an effort to keep seniors healthy. Taking care of people when they're sick is important, but Humana has decided that seniors in particular have a strong interest in maintaining their health, and it is rolling out a new initiative aimed at helping them do just that. The program offers a mix of opportunities related to cognitive function, physical fitness, and nutrition, but there is also a strong component geared toward advocacy. Humana is hoping that the payoff will come in the form of improved health and reduced claims costs. PMID- 17120860 TI - Recalibrating prelicensure education. PMID- 17120861 TI - A Delphi study of innovative teaching in baccalaureate nursing education. AB - The purpose of this Delphi study was to describe the process of innovative teaching in the baccalaureate nursing classroom, based on the consensus of expert nurse educators (n = 28). Round 1 involved a semi-structured survey that required participants to identify essential components of, facilitators of, and barriers to innovative teaching. Content analysis of the responses was used to construct the 62-item Round 2 survey in which participants ranked the importance of each item using a 7-point, Likert-type scale. The Round 3 survey included the same items and provided the median scores of the participants' own and other experts' responses. The highest-ranked essential components were faculty open, seeking new ideas; faculty motivation, commitment, and enthusiasm; and open and effective student-faculty communication. The highest-ranked facilitators were effective communication with students, faculty motivation and enthusiasm for change, and faculty satisfaction with innovative teaching. Faculty attitudes, fears, and lack of knowledge of innovative teaching were the highest-ranked barriers to innovative teaching in the nursing classroom. PMID- 17120862 TI - Reflection in the disability education of undergraduate nurses: an effective learning tool? AB - Reflection was integrated into an undergraduate nursing curriculum, with different frameworks demonstrating the application of a structured approach to reflection. Reflection is defined as examination and exploration of an issue of concern to help create or clarify meaning. The aim of this qualitative study was to evaluate the usefulness of formal reflection in the context of undergraduate nursing education during the disability module. Twelve reflection assignments written by second-year nursing students were analyzed. The analysis indicated that students' reflection focused less on their experience of working with people with disabilities and more on their overall learning experience and coping with clinical practice. A central theme, Coping with Clinical Practice, and four subthemes were identified. Students acknowledged reflection as beneficial to their learning and linked to their clinical practice. PMID- 17120863 TI - Use of a breathalyzer in a local bar: A community education project. AB - In completing a faculty-assigned service-learning project, a group of nursing students sought to educate their peers and the public about the effects of alcohol consumption. Breathalyzer readings were obtained from 150 bar patrons and correlated with the number of drinks consumed and number of hours spent drinking. The results were used to educate bar patrons about the difference between how intoxicated they felt and their actual blood alcohol levels. The students also distributed information about substance abuse assistance and safe transportation home. This screening technique served as an application of secondary prevention principles for the nursing students. The project was considered innovative in that no similar study was found in the literature. Selected Healthy People 2010 goals were also addressed in this project. PMID- 17120864 TI - The art of nursing expressed in poetry. AB - Writing is an essential skill for professional nurses. During their education, nursing students are required to produce a variety of written assignments, but most of these are regimented, such as care plans and term papers. There is little opportunity for students to express their creative side. Because all writing experiences are useful for student growth, this project in writing poetry was developed to provide students with an opportunity to reflect on their clinical experience in home health nursing and to document the contribution of clinical experience to their affective domain. PMID- 17120865 TI - Teaching therapeutic communication VIA camera cues and clues: the video inter active (VIA) method. AB - For nursing and other health care students to be effective, remain in their training, and subsequently join the health care workforce, they need to have good therapeutic communication skills. This article presents an innovative strategy to improve therapeutic communication skills for nursing and health sciences students. The Video Inter-Active (VIA) computer-based, challenge-response-record evaluate method used in this pilot study engaged students in realistic responses to scenarios commonly encountered in health care settings and provided them with opportunities to see themselves and critique their responses. Findings from this pilot study indicate that the VIA method is an effective and efficient way for students to develop and practice therapeutic communication skills. In a relatively short period (5 weeks), their learning from this method far PMID- 17120866 TI - Using audience response technology for pretest reviews in an undergraduate nursing course. AB - Implementing strategies to promote classroom interaction has been shown to enhance learning. In this study, we used an audience response system (ARS), an emerging technology, to actively engage students in examination reviews in an undergraduate nursing anatomy and physiology course. The reviews, set up as a pre test of 25 questions in a format similar to that of the game Jeopardy!, were used before six of the eight examinations. Average scores of the class on examinations preceded by the interactive ARS review were compared with those on examinations preceded by the more traditional, lecture-style review; no significant improvement due to ARS use was found. However, results from a student satisfaction survey about ARS use demonstrated positive attitudes and perceived benefit from this interactive technology. PMID- 17120867 TI - Using the interpretive narrative research method in interdisciplinary research projects. AB - Researchers from different disciplines bring to a project their different perspectives of the research problem. Differences in education, experiences, and understanding create a research project that has more depth and breadth than one designed by researchers from a single discipline. The purpose of this article is to describe how faculty from two disciplines, nursing and education, used the interpretive narrative method, a qualitative research method, in a pilot project to examine issues related to the recruitment and retention of Hispanic nurses. The researchers chose the interpretive narrative method for the flexibility it offers interdisciplinary research, its power in eliciting comprehensive narratives from participants, and the possibilities it offers for analysis. PMID- 17120868 TI - Influenza vaccination administration by junior and senior nursing students. PMID- 17120869 TI - Public health in a globalized world: breaking down social, economic and political barriers. PMID- 17120870 TI - Summarizing health inequalities in a Balanced Scorecard. Methodological considerations. AB - The association between social determinants and health inequalities is well recognized. What are now needed are tools to assist in disseminating such information. This article describes how the Balanced Scorecard may be used for summarizing data on health inequalities. The process begins by selecting appropriate social groups and indicators, and is followed by the measurement of differences across person, place, or time. The next step is to decide whether to focus on absolute versus relative inequality. The last step is to determine the scoring method, including whether to address issues of depth of inequality. PMID- 17120871 TI - Obesity rates among rural Ontario schoolchildren. AB - BACKGROUND: The majority of existing studies of obesity risk among Canadian children come from urban populations. The purpose of this study is to assess the prevalence of obesity in a sample of rural Ontario children. METHODS: Measures of height and weight were obtained for 504 children attending seven public elementary schools in Grey and Bruce Counties, a predominantly rural area of Southern Ontario. Body mass index (BMI, or weight/height2) scores were calculated and compared with reference data from the Centers for Disease Control. RESULTS: Rates of overweight and obesity were high in this sample, with 17.7% of children classified as overweight and 10.9% classified as obese. There was a significantly high prevalence of overweight for both boys (17.8%) and girls (17.5%) (Chi-square = 75.70, p < 0.001). However there was a significant gender difference in obesity prevalence: 15.0% of boys were obese, compared with 6.8% of girls (Mann-Whitney U = 29133.0, p > 0.05). CONCLUSION: Findings indicate that among rural children- particularly boys--risk of overweight and obesity are at least as high as in their urban Canadian counterparts. There appear to be fewer girls than boys at the extreme high end of the distribution of BMI, which may indicate differences in the growth environment of rural boys and girls. PMID- 17120872 TI - School region socio-economic status and geographic locale is associated with food behaviour of Ontario and Alberta adolescents. AB - BACKGROUND: In an attempt to elucidate broader determinants of adolescent dietary intake and habits, food intakes and selected food behaviours of grades 9 and 10 students from Ontario and Alberta were examined according to school region socio economic status and urban/rural locale. METHODS: Using a stratified random sample framework, 53 high schools from 28 school boards were recruited (45 public and 8 private; 33 urban and 20 rural). Median family income for Canada Post's forward sortation area of the school was used to define school region SES. Public and private schools were compared as a proxy measure of SES. A web-based survey of food intake and behaviours, including a 24-hour diet recall and food frequency questionnaire, was completed by 2,621 students in grades 9 and 10. Comparison of intakes and behaviours by school designation as urban/rural, public/private or regional SES (generalized linear model procedure) controlled for student gender and grade distribution and number of participants within schools. RESULTS: School region SES ranged from dollars 40,959 to dollars 85,922/year. Vegetable and fruit consumption (p < 0.001), fibre intake (p < 0.001) and frequency of breakfast consumption (p < 0.01) increased with increasing income, while added sugar intake decreased (p < 0.01). Private versus public school students had lower intakes of sweetened drinks (p < 0.01) and higher intakes of fibre (p=0.02). Rural students reported higher mean intakes of calcium (1106 vs. 995 mg/day, respectively, p = 0.03) and milk products (2.7 vs. 2.3 servings/day, p < 0.01) than urban students. CONCLUSION: Selected food behaviours of youth from Ontario and Alberta improve with increasing school SES and vary with rural/urban school locale. Identifying regional demographics may be useful in tailoring healthy eating programs to the specific school. PMID- 17120873 TI - Portrait of outpatient visits and hospitalizations for acute infections in Nunavik preschool children. AB - OBJECTIVE: Inuit children from around the world are burdened by a high rate of infectious diseases. The objective of this study was to evaluate the incidence rate of infections in Inuit preschool children from Nunavik (Northern Quebec). METHODS: The medical chart of 354 children from a previously recruited cohort was reviewed for the first five years of life. All outpatient visits that led to a diagnosis of acute infection and all admissions for acute infections were recorded. RESULTS: Rates of outpatient visits for acute otitis media (AOM) were 2314, 2300, and 732 events/1000 child-years for children 0-11 months, 12-23 months, and 2-4 years, respectively. Rates of outpatient visits for lower respiratory tract infections (LRTI) were 1385, 930, and 328 events/1000 child years, respectively. Rates of hospitalization for pneumonia were 198, 119, and 31 events/1000 child-years, respectively. CONCLUSION: Inuit children from Nunavik have high rates of AOM and LRTI. Such rates were higher than that of other non native North-American populations previously published. Admission for LRTI is up to 10 times more frequent in Nunavik compared to other Canadian populations. PMID- 17120874 TI - Seroprevalence of West Nile virus in Saskatchewan's Five Hills Health Region, 2003. AB - BACKGROUND: The Five Hills Health Region of Saskatchewan reported the highest West Nile virus (WNV) case rates in the 2003 outbreak. A serologic and telephone survey was undertaken to assess the seroprevalence of the virus and the knowledge, attitudes and behaviours of the residents. METHODS: Respondents had to be at least 18 years of age, and residents of the Five Hills Health Region between July 1st and September 15th, 2003. Blood samples of respondents were tested at the National Microbiology Laboratory for flavivirus immunoglobulin using a WNV IgG ELISA and plaque reduction neutralization test. Descriptive analyses performed related to respondents' demographics, knowledge, attitudes, behaviours, and seropositivity. WNV infection risk was assessed using odds ratio. RESULTS: There were 619 questionnaire respondents, of whom 501 donated a blood sample. The seroprevalence of WNV in the Five Hills Health Region was 9.98% (95% CI 7.37-12.59%). Seropositivity of rural areas was 16.8% and urban was 3.2%. Most (97%) of participants thought WNV was an important health issue. Forty-eight percent of the participants used insect repellents containing DEET most of the time. There was good knowledge regarding WNV transmission and prevention of the spread of WNV. Rural compared to urban residents were six times more likely to be positive for WNV (OR=6.13, 95% CI 2.82-13.34). INTERPRETATION: This is the highest seroprevalence rate of West Nile virus recorded in North America thus far. Many factors could have influenced this outbreak, such as eco-region, early prolonged hot weather, level of mosquito control programs, urban and rural community differences, and personal protective behaviours. PMID- 17120875 TI - Modelling geographic variations in West Nile virus. AB - BACKGROUND: This paper applies a method for modelling the spatial variation of West Nile virus (WNv) in humans using bird, environmental and human testing data. METHODS: We used data collected from 503 Alberta municipalities. In order to manage the effects of residual spatial autocorrelation, we used generalized linear mixed models (GLMM) to model the incidence of infection. RESULTS: There were 275 confirmed cases of WNv in the 2003 calendar year in Alberta. Our spatial model indicates that living in the grasslands natural region and levels of human testing are significant positive predictors of WNv; living in an urban area is a significant negative predictor. CONCLUSION: Infected bird data contribute little to our model. The variability of West Nile virus incidence in Alberta may be partly confounded by the variations in the rate of testing in different parts of the province. However, variation in infection is also associated with known environmental risk factors. Our findings are consistent with existing knowledge of WNv in North America. PMID- 17120876 TI - Shelter-based convalescence for homeless adults. AB - OBJECTIVES: Homelessness is associated with increased hospital costs and length of stay, and medical or surgical conditions are typically complicated by secondary diagnoses of substance abuse or mental illness. Convalescence care to provide timely treatment has not been analyzed. This is a retrospective study of diagnoses and utility of shelter-based convalescence in a cohort of homeless subjects. METHODS: A 20-bed shelter-based unit providing up to 3 months stay post hospital discharge, or for treatment of addictions or for those too ill to remain in the general shelter was studied. Charting was by the use of an electronic health record developed for the project. Demographics, reason for admission and outcomes are retrospectively described. RESULTS: 140 men had 181 admissions from July 2000-April 2003; 23.8% were post hospital discharge, 57.4% were from the general shelter. Average length of stay was 40 days. 83.4% were treated for a medical or surgical condition, 83.6% for psychiatric disease and 29.8% for addictions. Medication adherence was >80% in the majority. During admission, 20% obtained a new health card, 43.6% a new drug card, 89.3% received transportation to appointments, 60% applied for housing and 24.3% obtained housing. CONCLUSION: A shelter-based convalescence unit can provide health care to homeless persons, treat medical and mental illness, ensure adherence to treatment regimes, decrease substance abuse and assist with housing. PMID- 17120877 TI - Drug-related overdose deaths in British Columbia and Ontario, 1992-2004. AB - OBJECTIVE: To compare rates of fatal drug-related overdose death (OD) cases--a major harm outcome of illicit substance use--in the two provinces of British Columbia (BC) and Ontario, and the two largest municipalities in those provincial jurisdictions, namely the cities of Vancouver and Toronto, between 1992 and 2004. METHODS: Provincial coroners' data of drug-related OD cases for the provincial jurisdictions of BC and Ontario, and the municipal jurisdictions of Vancouver and Toronto, are descriptively presented and compared. RESULTS: After drastic increases in the initial part of the observation period, OD rates in BC have been declining; moreover, due to major reductions of OD cases in Vancouver, the ratio of OD cases between Vancouver and the province of BC has fallen considerably. Conversely, OD rates in Ontario have remained stable at low levels, whereas Toronto has seen a slight decline in such rates during the observation period. INTERPRETATION: The recent establishment and expansion of treatment and harm reduction interventions may have influenced the decline of ODs in BC, yet similar interventions in Ontario did not have a similar effect, perhaps due to different patterns of illicit drug use. OD rates in jurisdictions across Canada need to be monitored and analyzed to inform evidence-based policy development. PMID- 17120878 TI - Substance use among women in shelters for abused women and children. Programming opportunities. AB - OBJECTIVE: This study explores changes in the use of alcohol and other substances by women in British Columbia as they moved into shelters for abused women and again three months later. We see this time as a key life transition, and potentially a rich opportunity for influencing women's substance use behaviour. The purpose of this study was to document changes in the level of use of alcohol and other substances and the levels of stress among women as they moved through shelters for abused women. METHODS: Standardized questionnaires augmented by qualitative interviews were employed to measure alcohol and substance use, experiences of abuse, and levels and types of stressors facing women in this situation. FINDINGS: Significant reductions in women's use of alcohol and stimulants were observed from Interview I to Interview II, but there was no significant reduction in use of other depressants or tobacco use. Levels of stress decreased and sources of stress changed for the women after the shelter experience. Stress connected to relationship with partners had the most significant decrease, followed by mental health, housing, and legal issues. Women reported barriers to accessing financial aid and services for substance use outside of the shelter. CONCLUSIONS: Women's experiences of violence and substance use were found to be interconnected in complex ways and changes in substance use were affected by a range of influences, such as financial concerns, mothering, relationships, levels of social support, and physical and mental health issues. Substance-using women who have experienced violence are an underserved population and a multi-sectoral response designed to address psychosocial, relational, and structural issues could better help them improve their overall health. PMID- 17120879 TI - Bayesian small area cluster analysis of neural tube defects in Newfoundland. AB - BACKGROUND: The incidence of neural tube defects (NTDs) is declining worldwide due to the implementation of folic acid supplementation programs. Such a program was implemented over 1996-97 in Newfoundland and Labrador, Canada. The geographical distribution of birth incidence was studied prior to and after the implementation of the program to identify regions of residual high incidence. Excess residual cases may potentially be due to genetic causes or incomplete supplementation program implementation. METHODS: Maternal place of residence for all provincial live birth and stillbirth notifications, provincial maternal-fetal medicine referrals, provincial rehabilitation referrals, and all provincial hospitals with NTDs or terminations for NTDs was obtained from 1975 to 2002 for near complete case ascertainment. Bayesian small area analysis was separately performed on cases from 1975-1996 and 1997-2002. The two time periods were compared. RESULTS: Birth incidence of NTDs was noted to decline after 1996, from 5.54/1000 live births to 1.08/1000 live births. 592 cases were found from 1975 1996 and 34 cases from 1997-2002. Relative risk of birth incidence was 0.93-1.18 (95% CI) for 1975-1996 and 0.97-1.02 for 1997-2002 after Bayesian smoothing. One region had an excess of residual cases greater than 34%. CONCLUSIONS: The implications of this observation to the management of the public health initiative imply that overall response to the decrease in cases tends to be uniform across the province, with potentially one area of interest where extra efforts may be devoted. PMID- 17120880 TI - The three networks framework to deal with public health emergencies in Guangxi, China. AB - OBJECTIVE: To improve the detection and control of infectious diseases in Guangxi, China. SETTING/PARTICIPANTS: Guangxi province in southwest China has almost 50 million people, of whom approximately 30% reside in urban and 70% in rural areas. There are 12 minority nationality groups living in the region. INTERVENTION: A village doctor reports any infectious disease outbreak to the Reporting Network, which notifies the Service Network to organize the clinical response. This is supported by the Government Network that coordinates the response among the multiple layers of local governments. OUTCOMES: Since 2002 when the Three Networks system was first started, the time from incidence to report has been shortened on average from 30.6 to 7.6 days and the number of cases has increased from slightly less than 5000 cases/year (4965) to almost 10,000 cases/year (9873). Average mortality has decreased from 3.23% to 0.74%. The Three Networks system has been successful in controlling measles outbreaks; and during SARS, when 11 cases came to Guangxi from the neighbouring Guangdong province, there were only 11 additional new cases with no community spread and no spread to medical staff. CONCLUSION: The Three Networks system has played an important role in infectious disease prevention and control in Guangxi province, and may be applicable to other areas with a similar situation. PMID- 17120881 TI - The promise of public health: ethical reflections. AB - Public health is in the spotlight of public and political concern, providing a unique window of opportunity for its revitalization and restoration as a pillar of the Canadian health care system. The establishment of a Federal Public Health Agency is a critical first step. The public health community has identified key challenges toward this renewal. However, public health ethics have received little attention and, when addressed, have focussed almost exclusively on communicable diseases. The ethical issues inherent in public health transcend infectious diseases and are distinct from clinical and research ethics. Identifying and addressing ethical issues at the heart of public health, including the public interest and the common good, as well as fundamental issues related to the core functions of public health will be essential if this revitalization of public health is to be achieved. While legal and regulatory reforms are necessary, they will not be sufficient to adequately address fundamental questions of the valuing of public health or the ethical issues in public health. Elements of a research agenda on the ethical foundations of public health reform are identified here. PMID- 17120882 TI - Three challenges to the Ottawa Spirit of Health promotion, trends in global health, and disabled people. AB - Health promotion according to the 1986 Ottawa Charter of the first global health promotion conference "is the process of enabling people to increase control over, and to improve, their health. To reach a state of complete physical, mental and social well-being, an individual or group must be able to identify and to realize aspirations, to satisfy needs, and to change or cope with the environment". In this commentary, I explore three powerful challenges to the spirit of the Ottawa Charter and to global health. The first challenge is the departure from the WHO definition of health; the second challenge relates to the appearance of the transhumanist/enhancement model of health which includes human performance enhancement beyond species-typical boundaries as part of the concept of health. The third challenge consists of the limited involvement and understanding of disabled people with their different models of 'disability/impairment' (medical, social, transhumanist/enhancement) in the discourse of global health and health promotion. Not dealing with these challenges impairs the ability of health promotion to deal with global health problems, the 'health' needs of marginalized groups--in particular, disabled people--and the Millennium Development Goals. PMID- 17120883 TI - Primary care (PC) and primary health care (PHC). What is the difference? AB - Primary Care and Primary Health Care are very similar terms which are often employed interchangeably, but which are also used to denote quite different concepts. Much time and energy is spent discussing which term is the appropriate one for a particular application. There is a growing recognition internationally that the two terms describe two quite distinct entities. Recent Canadian uses of the two terms are, for the most part, consistent with the international uses. Primary Care, the shorter term, describes a narrower concept of "family doctor type" services delivered to individuals. Primary Health Care is a broader term which derives from core principles articulated by the World Health Organization and which describes an approach to health policy and service provision that includes both services delivered to individuals (Primary Care services) and population-level "public health-type" functions. PMID- 17120884 TI - The problem with breastfeeding discourse. AB - This paper reflects on how informational biases have subtly entered into the breastfeeding health discourse and recommends how focus might be redirected. Specifically, contemporary infant feeding health discourse reflects a biased representation of the scientific literature, and fails to provide an appropriate contextualization of risk and benefit. It is important to correct these biases in order to uphold the foundational ethics of expert-guided childcare, and to rightly reposition the discourse as a tool for influence via education, not informational manipulation. Correcting these biases would also ease the discursive pressure and associated emotional burdens mothers currently experience in relation to infant feeding, and would reinforce a needed commitment to the development of appropriate supports to help mothers viably exercise their right to breastfeed. PMID- 17120885 TI - [Interdisciplinary health research]. AB - Health care professionals are increasingly called upon to work collaboratively. This commentary discusses our experience at the Summer Institute on Interdisciplinary Health Research, which aimed to increase the capacity of future Canadian researchers to work with health professionals from other disciplines. We discuss, in particular, our small group experience--the challenges we faced, and how we were able to resolve the internal conflicts that occurred. We also discuss the conditions necessary to engage in interdisciplinary work and offer suggestions to improve future initiatives to build interdisciplinary research capacity. PMID- 17120886 TI - Pathognomonic radiological signs for predicting prognosis in patients with chronic atlantoaxial rotatory fixation. AB - OBJECT: The authors conducted a study to assess the correlation between radiological features and clinical courses in patients with chronic atlantoaxial rotatory fixation (AARF) and to determine diagnostic imaging signs for predicting prognosis. METHODS: There were 24 patients (eight boys and 16 girls) whose mean age was 7.8 years (range 4-14 years) and in whom AARF was diagnosed. The mean follow-up duration was 3.7 years (range 5 weeks-12 years). There were two groups of patients: 15 patients who visited the hospital less than 8 weeks after symptom (torticollis) onset comprised the acute group, and nine patients in whom the diagnosis of AARF was established later than 3 months after symptom onset comprised the chronic group. The chronic group was divided into two subgroups: those in whom the closed reduction could be achieved and those in whom it could not. Clinical data and radiographic studies, including plain radiographs and plain and three-dimensional (3D) computed tomography (CT) reconstructions, were reviewed retrospectively. A deformity of the superior C-2 facet joint was frequently observed in the group of patients with chronic AARF (p < 0.0001). This sign represented a risk factor for recurrent dislocation (p = 0.0003, Fisher exact test). Prominent lateral inclination of C-1 was an impeding factor for reduction of chronic AARF (p < 0.0001, analysis of variance with Fisher post hoc test). Greater than 20 degrees of lateral inclination of the atlas indicated an irreducible subluxation (p = 0.0023, Fisher exact test). CONCLUSIONS: Both facet joint deformity and lateral inclination observed on 3D CT reconstructions can be useful signs to predict the prognosis and the treatment of choice in patients with chronic AARFs. PMID- 17120887 TI - Cervical spine epidural abscess: experience with microsurgical treatment in eight cases. AB - OBJECT: The authors report a series of eight consecutive cases in which epidural abscesses in the cervical spine were treated by microsurgery without arthrodesis, including two cases of concomitant pyogenic and tubercular infection. METHODS: The authors used a minimally invasive surgical approach consisting of single level anterior microsurgical discectomy and drainage of the epidural abscess via a silicone catheter, and then initiated antibiotic therapy. At follow-up examination (mean duration 39 months), six patients exhibited complete recovery and two suffered from minor residual deficits. In all cases, spontaneous vertebral fusion occurred. Sagittal alignment was maintained in seven patients, and in one there was slight asymptomatic kyphosis. In two patients, tubercular and pyogenic infections were found. Prior intervention for dental infection was recorded in four cases. CONCLUSIONS: In the absence of preoperative spinal instability, microsurgical drainage of the abscess followed by specific antibiotic therapy resulted in spinal cord decompression and neurological recovery, thereby facilitating spontaneous fusion and vertebral stability. The presence of combined tubercular and pyogenic infections of the cervical spine should be considered, especially in patients whose immune systems are depressed. PMID- 17120888 TI - Facet cyst in the lumbar spine: radiological and histopathological findings and possible pathogenesis. AB - OBJECT: The authors define facet cyst as a cyst located beside the facet joint and exhibiting a communication with the joint, as demonstrated on arthrography and subsequent computed tomography (CT) of the joint space. The purpose of this study was to determine the pathogenesis of facet cysts based on their radiological and histological features. METHODS: Forty-six juxtafacet cysts in 45 patients (26 men and 19 women, age range 41-82 years) were surgically treated after evaluation by arthrography and subsequent CT scanning. A communication channel between the cyst and the joint was confirmed in all cases and thus the lesions were designated facet cysts. In almost all patients the involved facet joint showed moderate to severe degeneration. After a thorough preoperative radiological evaluation, these cysts were excised en bloc by medial facetectomy with the entire ligamentum flavum. The specimens were cut axially at the maximum diameter and were histologically investigated. Morphologically, the cysts exhibited three shapes, appearing as: 1) a small protrusion, 2) a semicircular cyst, or 3) a round cyst. The cyst walls consisted of elastic and collagen fibers undergoing fibrinoid degeneration, but no synovial lining cells were detected. The cystic cavities were regularly filled with fibrinoids, and myxoid degeneration was found particularly in the larger cysts. CONCLUSIONS: Facet cysts are closely related to the degeneration of the neighboring facet joint. Analysis of the findings in this histological study suggests that there are several shapes of facet cysts. The authors propose that the shape could depend on the stage of the cyst's development. PMID- 17120889 TI - Evaluation of postoperative spinal epidural hematoma after microendoscopic posterior decompression for lumbar spinal stenosis: a clinical and magnetic resonance imaging study. AB - OBJECT: The incidence of postoperative spinal epidural hematoma (SEH) is low, and to the best of the authors' knowledge, no researchers have evaluated its actual incidence and clinical features. The purpose of this study was to investigate the clinical consequences of SEH after microendoscopic posterior decompression (MEPD) in patients with lumbar spinal stenosis. METHODS: Data obtained in 30 patients undergoing MEPD for lumbar spinal stenosis were reviewed. At 1 week after surgery, magnetic resonance (MR) imaging documented SEHs in 10 patients (33% [Group 1]) and no evidence of SEHs in 20 patients (67% [Group 2]). The authors compared MR imaging findings, postoperative morbidities, and clinical outcomes between the groups. Three Group 1 patients had symptomatic SEHs. All symptoms were mild without associated neurological deterioration and spontaneously subsided within 3 weeks of surgery. Magnetic resonance imaging demonstrated spontaneous regression of the SEH in all patients at 3 months after surgery. In Group 1 patients, however, the authors observed less expansion of the dural sac after 1 year despite sufficient widening of the osseous spinal canal. Low-back pain within 1 week of surgery was moderate in Group 1 and mild in Group 2. Improvements at the final follow up were greater in Group 2 patients. CONCLUSIONS: The incidence of postoperative SEHs may be greater than reported. Postoperative SEHs caused poor expansion of the dural sac despite its spontaneous regression. In addition, postoperative SEHs caused a delay in the patient's recovery and led to a poor clinical improvement. The prevention of postoperative SEHs might be required to prevent not only neurological deterioration but also a delay in the patient's recovery. PMID- 17120890 TI - Incidence of unusual and clinically significant histopathological findings in routine discectomy. AB - OBJECT: Routine histopathological examination of discectomy specimens remains common practice in many hospitals, although it rarely detects unsuspected clinically significant disease. Controversy exists as to the effectiveness of this practice. The objectives of this study were to compare the authors' experience with a review of the literature. METHODS: In a retrospective database analysis the authors identified all intervertebral disc specimens obtained during spinal procedures over an 8-year period (1996-2004). Cases of benign (nonneoplastic and noninfectious) indications for surgery were included in the study, whereas cases of nonbenign indications were excluded. The final pathological diagnoses were reviewed, and a chart review was performed to determine whether any unexpected findings affected subsequent patient care. A total of 1858 discectomy specimens were identified: 1775 of these were obtained in 1719 routine discectomy procedures. Unexpected histopathological findings were identified in four cases, and none was clinically significant. CONCLUSIONS: Routine histopathological examination of disc specimens is not justified. The decision to send specimens for pathological examination should be determined on a case-by-case basis after consideration of the clinical presentation, results of laboratory and imaging studies, and intraoperative findings. PMID- 17120891 TI - Surgical treatment of lumbar epidural varices. AB - OBJECT: An epidural varix of the lumbar spine is an acquired pathological entity that results from dilation of the internal vertebral venous plexus. The dilated veins in the epidural spaces or in the intervertebral foramen cause irritation or compression of the thecal sac and nerve roots, and this in turn causes lumbar radiculopathy. The lesions develop as an independent process or are accompanied by degenerative changes of the lumbar spine. Resection of varicose veins can be complicated when significant venous bleeding occurs. The purpose of the present study was to develop diagnostic criteria for lumbar epidural varices and methods of surgical treatment and to analyze the results of treatment. METHODS: There were 43 surgically treated patients in whom lumbar radiculopathy was caused by epidural varices. The mean duration of symptoms was 11 months (range 5-49 months) and the mean age of the patients was 41 years (range 26-68 years). Segmentary varices were found in 14 patients, local varices in 21, and extensive varices in eight. Patients underwent preoperative magnetic resonance imaging and vertebral venography. Of 43 cases, the varicose veins were completely occluded in all 14 patients with segmentary varices of the segmentary-type lesion. Of 21 patients with local varices, total occlusion was obtained in 17 patients and partial occlusion in four. In the patients with extensive varices, total occlusion was not performed; instead, partial occlusion was undertaken. Resolution of lumbar radicular syndromes was observed, to some extent, in all patients. Radicular pain regressed in 12 of 14 patients with segmentary varices, in 18 of 19 patients with local varices, and in three of four patients with extensive varices. Nerve root related sensory disturbances regressed in six of 10 patients with segmentary varices, in 11 of 15 patients with local varices, and in one patient with extensive varices. Motor deficits regressed in three of eight patients with segmentary varices, in three of six patients with local varices, and in none among those with extensive varices. CONCLUSIONS: Epidural varicosity is a specific form of spinal disease. Its diagnostic clarification and surgical treatment are difficult and have only recently been developed. Precise diagnosis and a well-planned surgery allow for a positive outcome. PMID- 17120892 TI - The use of hemopoietic stem cells derived from human umbilical cord blood to promote restoration of spinal cord tissue and recovery of hindlimb function in adult rats. AB - OBJECT: The use of human umbilical cord blood (HUCB) cells has been reported to improve functional recovery in cases of central nervous system injuries such as stroke, traumatic brain injury, and spinal cord injury (SCI). The authors investigated the effects of hemopoietic stem cells that were derived from HUCB and transplanted into the injured spinal cords of rats. METHODS: One week after injury, an HUCB fraction enriched in CD34-positive cells was transplanted into the experimental group. In control animals, vehicle (Matrigel) was transplanted. Recovery of motor functions was assessed using the Basso-Beattie-Bresnahan Locomotor Scale, and immunohistochemical examinations were performed. Cells from HUCB that were CD34 positive improved functional recovery, reduced the area of the cystic cavity at the site of injury, increased the volume of residual white matter, and promoted the regeneration or sparing of axons in the injured spinal cord. Immunohistochemical examination revealed that transplanted CD34-positive cells survived in the host spinal cord for at least 3 weeks after transplantation but had disappeared by 5 weeks. The transplanted cells were not positive for neural markers, but they were positive for hemopoietic markers. There was no evidence of an immune reaction at the site of injury in either group. CONCLUSIONS: These results suggest that transplantation of a CD34-positive fraction from HUCB may have therapeutic effects for SCI. The results of this study provide important preclinical data regarding HUCB stem cell-based therapy for SCI. PMID- 17120893 TI - Effect on bone induction of using contrast media to reconstitute recombinant human bone morphogenetic protein-2 in an ectopic model in rats. AB - OBJECT: In this study the authors tested the osteoinductive potential of recombinant human bone morphogenetic protein-2 (rhBMP-2) when combined with each of three commercially available contrast media (Conray, Omniscan, and Optiray). METHODS: Initial in vitro and cadaver tests verified the feasibility of using contrast media to visualize absorbable collagen sponge implants containing rhBMP 2 on fluoroscopic radiographic images. For the feasibility studies, lyophilized rhBMP-2 was prepared for injection by reconstitution with contrast media instead of sterile water. For the in vivo study, samples of an rhBMP-2 stock solution were diluted to 0.1 mg/ml by using three contrast media. In each sample, the final solution consisted of 97% contrast medium by volume. Recombinant human bone morphogenetic protein-2 diluted with sterile water for injection was used as a positive control. The rhBMP-2 solutions were applied to 0.5-cm3 collagen sponges and implanted subcutaneously on the thoracic cavity of athymic rats. At 4 weeks, the rats were killed, and the implants were removed. The explants were graded for degree of bone formation by using manual palpation and radiographic and histological assessments. CONCLUSIONS: By all methods of evaluation used, rhBMP-2 diluted with Omniscan was equivalent to rhBMP-2 diluted with sterile water in inducing bone formation. Both Conray and Optiray were shown to inhibit the osteoinductive potential of rhBMP-2. PMID- 17120894 TI - Anatomical relationships between the V2 segment of the vertebral artery and the cervical nerve roots. AB - OBJECT: During surgical procedures focused on the cervical nerve roots, the surgeon works in proximity to the V2 segment of the vertebral artery (VA). Depending on the specific surgical approach, it may be necessary to identify, expose, or mobilize the artery. In most cases, the artery may be left undisturbed. To reduce the risk of iatrogenic injury to the V2 segment during anterior and anterolateral approaches to the cervical spine, the authors analyzed the relationship between the V2 segment and the proximal segment of the C3-6 nerve roots. METHODS: Six cadaveric cervical spines (12 sides) were fixed with formalin, injected with red and blue latex, and investigated intraoperatively using different magnifications (x 3-40). The VA rested on the anteromedial surface of the cervical nerve roots at the level of each intertransverse space. The exiting nerve roots intersected the VA at a distance ranging from 4.5 to 8.1 mm (mean 6.3 +/- 1.06 mm) from the dural sac. The distance was slightly shorter at cephalad levels, suggesting that the artery is more posteriorly and medially situated at those levels. Arterial pedicles anchored the VA to the cervical nerve roots at various levels. These arteries gave rise to purely radicular, ligamentous, and medullary branches without a predictable pattern. After reaching the nerve roots on their lower margin, the nonligamentous branches pierced the radicular dural sheath within the neural foramen at a distance of 2 to 4 mm from the VA. CONCLUSIONS: Proximal-to-distal dissection of a cervical nerve root may proceed with relative safety for at least 4 mm. The V2 segment of the VA gives rise to at least one radicular arterial pedicle between C-4 and C-6. These trunks give rise to purely radicular, ligamentous, and medullary branches in an unpredictable pattern. PMID- 17120895 TI - Giant sacral meningeal diverticula: surgical implications of the "thecal tip" sign. Report of two cases. AB - The surgical anatomy of giant sacral meningeal diverticula varies greatly depending on whether they develop ventral or dorsal to the thecal sac and spinal nerve roots. The ability to distinguish between the two lesion types preoperatively is therefore advantageous. The authors present a method of distinguishing ventral from dorsal meningeal diverticula on magnetic resonance imaging using the "thecal tip sign." They also describe the differences in operative technique required for resection of each type of diverticular cyst. PMID- 17120896 TI - Type A intradural spinal arteriovenous fistula. Case report. AB - In this report, the authors present the case of a patient with a unique type of spinal arteriovenous fistula. Both the location and venous angioarchitecture of this variant are uncommon, making diagnosis of the lesion challenging and raising particular management issues. The authors discuss this unusual lesion and describe its imaging features and surgical findings, as well as highlight its pathological abnormalities. PMID- 17120897 TI - Tumoral calcinosis in bilateral facet joints of the lumbar spine in scleroderma. Case report. AB - Tumoral calcinosis commonly occurs in the articular soft tissues of the extremities but rarely in the spine. The authors performed surgery to treat lumbar tumoral calcinosis in a patient with scleroderma, in whom symptoms of neurological dysfunction had manifested. This 49-year-old woman presented with low-back pain and gait disturbance. Seven years before presentation, scleroderma had been diagnosed, and the patient had received medical treatment ever since. Imaging revealed tumoral calcinosis centered at the bilateral facet joints between L-3 and L-4, marked stenosis of the spinal canal, L-3 spondylolisthesis, and intervertebral instability. Surgery was performed to excise the lesion en bloc. After neural decompression, posterolateral fusion and pedicle screw fixation were undertaken. Symptoms improved after surgery. In this case, the underlying scleroderma that predisposes to calcinosis and facet joint degeneration due to lumbar spondylolisthesis were probably factors leading to the development of tumoral calcinosis in the lumbar spine. PMID- 17120898 TI - Regression of retroodontoid pseudotumors following C-1 laminoplasty. Report of three cases. AB - The authors report a new technique for C-1 laminoplasty without fusion in the treatment of cervical myelopathy associated with a retroodontoid pseudotumor (also known as a phantom tumor). The authors review the cases of three patients who underwent C-1 laminoplasty in which hydroxyapatite was used and fusion was not performed. All patients suffered from severe progressive myelopathy before surgery. Magnetic resonance imaging revealed a retroodontoid pseudotumor compressing the spinal cord at the C-1 level in all cases. Computed tomography was performed to examine the extent of bone erosion at the atlantoaxial joint and dens. Clinical parameters included neurological function, measured using the Japanese Orthopaedic Association score, and neck pain. Imaging parameters included pre- and postoperative atlas-dens intervals; the space available for the spinal cord; instability of the atlantoaxial joints; osteoarthritic changes of the atlantoaxial joint; postoperative changes in T2 high-intensity signal; and postoperative alteration in the size of the pseudotumor. Magnetic resonance imaging demonstrated complete disappearance of the pseudotumor in two cases and partial reduction in one case. In all three cases sufficient neurological recovery was observed. The mean recovery rate was 87.0%. This new technique is less invasive than standard procedures, preserves the cervical range of motion, and avoids the morbidity of obtaining a bone graft and placing instrumentation. The authors conclude that C-1 laminoplasty without fusion is an option in the surgical management of cervical myelopathy associated with a retroodontoid pseudotumor, either without C1-2 instability or with slight, but reducible, C1-2 instability. PMID- 17120899 TI - Endoscopic vertebroplasty for the treatment of chronic vertebral compression fracture. Technical note. AB - The authors describe a new vertebroplasty technique for the treatment of chronic painful vertebral compression fractures (VCFs). A urinary balloon catheter is introduced into the vertebral body (VB) via a bilateral transpedicular approach and inflated with contrast medium to obtain sufficient space for endoscopic observation. The granulation tissue occupying the VB is then removed using a punch or curette inserted through one pedicle, with the guidance of an endoscope introduced through the contralateral pedicle. After endoscopic resection of granulation tissue in the fractured VB, vertebroplasty is performed by injecting calcium phosphate cement (CPC) into the VB. Fourteen patients in whom chronic painful VCFs were diagnosed underwent surgery involving the aforementioned technique. In all cases, intractable pain and ambulatory function improved after surgery, and there were no significant systemic complications. On radiological evaluation in eight cases in which the follow-up period exceeded 1 year, the mean height of the fractured VB improved from 38% of that of adjacent intact VBs to 85%. Although a slight loss of correction was routinely observed at 1 month postoperatively, an additional loss of VB height was not noted up to 1 year later. Bone formation was commonly seen along the anterior wall of the involved vertebrae in all cases. Vertebroplasty involving the endoscopic removal of granulation tissue proved to be an efficacious procedure for the treatment of chronic painful VCFs. The osteoconductive capacity of CPC facilitated callus formation and ultimately restoration of vertebral bone structure. PMID- 17120900 TI - Dynamic retraction of the psoas muscle to expose the lumbar spine using the retroperitoneal approach. Technical note. AB - The retroperitoneal surgical approach has gained acceptance as a way to access the ventral aspect of the lumbar spine. Visualization is often limited, however, by the psoas muscle, which lies along the posterolateral aspect of the spine. Improved visualization is often attempted by retracting the muscle from the wound, which generally pulls the muscle laterally from the spine but not posteriorly, which is desirable for a better exposure of the spine, particularly the neural elements. In this paper, the authors describe a simple, atraumatic technique for retraction of the psoas muscle that allows excellent visualization of the spine. PMID- 17120901 TI - Complete removal of an intraspinal and extraspinal cervical chordoma in one stage using the lateral approach. Technical note. AB - The authors describe a precise surgical technique in which a large intraspinal and extraspinal, multivertebral, cervical chordoma was completely removed in one stage using the lateral approach. The patient in this case was a 29-year-old woman who presented with signs of radicular pain in the left C-3 area. Computed tomography and magnetic resonance imaging demonstrated a large intra- and extraspinal multivertebral tumor from C-2 to C-5, a finding that suggested a cervical chordoma. The tumor was completely removed in one stage using the lateral approach while controlling the vertebral artery (VA), and a partial corporectomy of C2-5 was also performed. Results from a postoperative histopathological examination confirmed that the tumor was a typical chordoma. The patient's postoperative course was uneventful. Cervical chordomas are typically excised using a posterior-anterior surgical approach with partial resection of the tumor. The lateral approach was appropriate in this patient for complete resection in one stage, because it enabled the surgeons to control the VA and access both extraspinal and intraspinal components of the chordoma. PMID- 17120902 TI - Intradural extramedullary spinal anaplastic ependymoma. Case illustration. PMID- 17120903 TI - The V2 segment of the vertebral artery. PMID- 17120904 TI - The V2 segment of the vertebral artery. PMID- 17120905 TI - Cervical cage migration. PMID- 17120906 TI - Infinite possibilities. PMID- 17120907 TI - Bridging the gaps. PMID- 17120908 TI - GeoWall: stereoscopic visualization for geoscience research and education. PMID- 17120909 TI - Swordplay: innovating game development through VR. PMID- 17120910 TI - Simulators and closed interaction loops. PMID- 17120911 TI - Online remeshing for soft tissue simulation in surgical training. AB - To graphically model and animate the realistic behavior of deformable tissue in surgical simulations, the authors' system adapts tetrahedra resolution by dynamically retessellating the mesh in and around the regions of interest. This technique overcomes limitations of previous methods that made it difficult to modify the mesh's topology online. PMID- 17120912 TI - Liver surgery planning using virtual reality. AB - In liver surgery planning, 2D and desktop-based 3D systems offer surgeons limited assistance. By using VR technology to liberate 3D from 2D input devices such as the mouse and keyboard, this surgery planning system better supports surgeons. User studies show that the system is both effective and easy to use. PMID- 17120913 TI - Visuohaptic simulation of bone surgery for training and evaluation. AB - Visual and haptic simulation of bone surgery can support and extend current surgical training techniques. The authors present a system for simulating surgeries involving bone manipulation, such as temporal bone surgery and mandibular surgery, and discuss the automatic computation of surgical performance metrics. Experimental results confirm the system's construct validity. PMID- 17120914 TI - Real-time finite-element simulation of linear viscoelastic tissue behavior based on experimental data. AB - The lack of experimental data on the viscoelastic material properties of live organ tissues has been a significant obstacle in the development of realistic models. A real-time and realisti finite-element simulation of viscoelastic tissue behavior using experimental data collected by a robotic indenter offers one solution. PMID- 17120915 TI - Conceptual framework for laparoscopic VR simulators. AB - Availability of YR laparoscopic simulation for surgical training has increased significantly. Nevertheless, few studies have explored such simulators' requirements and the degree of fidelity necessary to provide effective educational tools. The authors aim to identify which didactic resources available in YR simulation technologies are most important for laparoscopic training. PMID- 17120916 TI - Modeling crowd and trained leader behavior during building evacuation. AB - This article considers animating evacuation in complex buildings by crowds who might not know the structure's connectivity, or who find routes accidentally blocked. It takes into account simulated crowd behavior under two conditions: where agents communicate building route knowledge, and where agents take different roles such as trained personnel, leaders, and followers. PMID- 17120917 TI - Seeing is believing: using computer graphics to enthuse students. PMID- 17120918 TI - How to solve a cubic equation, part 3: general depression and a new covariant. PMID- 17120919 TI - Redesigning health systems for quality: Lessons from emerging practices. AB - BACKGROUND: It has been five years since the Institute of Medicine (IOM) report, Crossing the Quality Chasm, proposed systemwide changes to transform our health care system. What progress has been made? What lessons have been learned? How should we move forward? METHODS: Semistructured telephone interviews were conducted with 16 health care providers and researchers at organizations involved in system redesign. The findings were supplemented with a focused literature review and discussions from a national expert meeting. RESULTS: Many promising and innovative examples of redesign were identified. However, even delivery systems that are redesigning care in pursuit of the six IOM aims face daunting challenges, reflecting the need to align system changes across multiple levels and to integrate redesign efforts with ongoing system features. Four success factors were reported by providers as crucial in overcoming redesign barriers: (1) directly involving top and middle-level leaders, (2) strategically aligning and integrating improvement efforts with organizational priorities, (3) systematically establishing infrastructure, process, and performance appraisal systems for continuous improvement, and (4) actively developing champions, teams, and staff. A framework that integrates these success factors to facilitate a systems approach to redesigning health care organizations and delivery systems for improved performance is provided. CONCLUSIONS: Successful system redesign requires coordinating and managing a complex set of changes across multiple levels rather than isolated projects. PMID- 17120920 TI - Eliminating nosocomial infections at Ascension Health. AB - BACKGROUND: Eliminating nosocomial infections was identified as one of eight priorities for action for Ascension Health. St. John Hospital and Medical Center (SJHMC), and St. Vincent's Hospital (STV), designated alpha sites, developed best practices for the prevention of catheter-related blood stream infections (CR BSIs) and ventilator-associated pneumonia (VAP), respectively. METHODS: Both hospitals used the Institute for Healthcare Improvement model of "bundles" to achieve the goal of reducing nosocomial infections and also implemented multidisciplinary rounds and the use of daily goal sheets in the intensive care unit (ICU). RESULTS: Through the use of ventilator bundle, central line (CL) bundle, MDRs, and daily goal sheets, both facilities reduced CR-BSIs and VAPs by more than 50%. DISCUSSION: SJHMC saw the benefit of having the physical presence of the ICPs in the ICUs, providing the staff with on-the-spot reinforcement of the initiative. STV found by starting the change process through the use of a flexible MDR team, the hospital was able to successfully implement positive changes in its ICU culture. On the basis of the success in the ICU, the concept of MDR teams eventually was adapted and spread to all units. Open communication among all patient caregivers was extended and served to provide improved patient care throughout the hospital. PMID- 17120921 TI - The 100,000 Lives Campaign: A scientific and policy review. AB - BACKGROUND: On June 14, 2006, the Institute for Healthcare Improvement (IHI) announced that its campaign to save 100,000 lives had far surpassed its goal--by saving 122,300 lives. THE INTERVENTIONS: Although many of the campaign's six "evidence-based practices" are supported by relatively strong evidence, the use of rapid response teams (the only intervention not already required or promoted by a major federal or Joint Commission initiative) is not. THE "LIVES SAVED": Secular trends could account for many of the "lives saved," which IHI acknowledges (and could have adjusted for, resulting in a markedly lower "lives saved" estimate). Moreover, IHI's estimates of lives saved are dependent on the case-mix adjustment-accounting for nearly three out of four "lives saved." The actual mortality data were supplied without audit by the more than 3,000 participating hospitals, and 14% of the hospitals submitted no data at all. POLICY IMPLICATIONS: IHI established and promoted a set of achievable goals for American hospitals and generated unprecedented amounts of social pressure for hospitals to participate. This remarkable achievement should be studied by other organizations seeking to generate widespread change in the health care field. CONCLUSIONS: Although the 100,000 Lives Campaign succeeded in catalyzing efforts to improve safety and quality in American hospitals, the promotion of rapid response teams as a national standard is problematic, and methodologic concerns regarding the "lives saved" calculations make it difficult to interpret the campaign's true accomplishments. PMID- 17120922 TI - IHI replies to "The 100,000 Lives Campaign: A scientific and policy review. PMID- 17120923 TI - The Codman competition: Rewarding excellence in performance measurement, 19 -20. AB - BACKGROUND: Since 1997 the Ernest Amory Codman Award, the only health care award that recognizes excellence in performance measurement, has honored organizations and individuals for their use of process and outcomes measures to improve organization performance and quality of care. INDIVIDUAL AWARD WINNERS: The individual Codman award winners have advanced measurement of systems performance, health outcomes, and customer satisfaction. ORGANIZATION AWARD WINNERS: Forty-two organizations have been selected as winners. The work for which these organizations were recognized was categorized as improvements in direct patient care services, improved effectiveness of care through better teamwork, interdisciplinary planning, improvement of administrative processes, and improved quality of care in large regions or health care systems. Case studies from four organizations that have won the Codman award each represents a lesson or theme that may be instructive for other health care organizations--(1) the need for catalysts or agents of change, (2) evidence-based clinical pathways are essential for delivering optimal care to patients in large organizations, (3) quality assessment and improvement methods from other industries can be successfully applied to health care, (4) as health care is increasingly delivered by large networks and systems, quality takes on regional and even national relevance. CONCLUSION: The scope of Codman's endeavors is reflected in the array of quality improvement projects selected as Codman award winners. PMID- 17120924 TI - The medical emergency team as a safety net. AB - In the case of an 11-year-old boy with stable physiological parameters, the deterioration in condition was so sudden that only a rapid response system could ensure any chance of survival. PMID- 17120925 TI - A model for building a standardized hand-off protocol. AB - BACKGROUND: The Joint Commission has made a "standardized approach to hand-off communications" a National Patient Safety Goal. METHOD: An interactive 90-minute workshop (hand-off clinic) was developed in 2005 to (1) develop a standardized process for the handoff, (2) create a checklist of critical patient content, and (3) plan for dissemination and training. CONCLUSION: To date, 7 of 10 residency programs have participated. Analysis of these protocols demonstrated that the hand-off process is highly variable and discipline-specific. Although all disciplines required a verbal handoff, because of competing demands, verbal communication did not always occur. In some cases, the transfer of professional responsibility was separated in time and space from the transfer of information. For example, in two cases, patient tasks were assigned to other team members to facilitate timely departure of a postcall resident (to meet resident duty-hour restrictions), but results were not formally communicated to anyone. The hand-off clinic facilitated the incorporation of "closed-loop" communication by requiring that follow-up on these tasks be conveyed to the on-call resident. DISCUSSION: This model for design and implementation can be applied to other health care settings. PMID- 17120926 TI - Fatty acid production in Schizochytrium sp.: Involvement of a polyunsaturated fatty acid synthase and a type I fatty acid synthase. AB - Schizochytrium sp. is a marine microalga that has been developed as a commercial source for docosahexaenoic acid (DHA, C22:6 (omega-3), enriched biomass, and oil. Previous work suggested that the DHA, as well as docosapentaenoic acid (DPA, C22:5 omega-6), that accumulate in Schizochytrium are products of a multi-subunit polyunsaturated fatty acid (PUFA) synthase (1). Here we show data to support this view and also provide information on other aspects of fatty acid synthesis in this organism. Three genes encoding subunits of the PUFA synthase were isolated from genomic DNA and expressed in E. coli along with an essential accessory gene encoding a phosphopantetheinyl transferase (PPTase). The resulting transformants accumulated both DHA and DPA. The ratio of DHA to DPA was approximately the same as that observed in Schizochytrium. Treatment of Schizochytrium cells with certain levels of cerulenin resulted in inhibition of 14C acetate incorporation into short chain fatty acids without affecting labeling of PUFAs, indicating distinct biosynthetic pathways. A single large gene encoding the presumed short chain fatty acid synthase (FAS) was cloned and sequenced. Based on sequence homology and domain organization, the Schizochytrium FAS resembles a fusion of fungal FAS beta and alpha subunits. PMID- 17120927 TI - Different kinetic in incorporation and depletion of n-3 fatty acids in erythrocytes and leukocytes of mice. AB - n-3 PUFA are well known for their anti-inflammatory effects. However, there has been only limited study on the kinetics of incorporation and depletion of n-3 PUFA in immune cells. In the present study we investigated the incorporation and depletion of n-3 PUFA in erythrocytes and leukocytes in mice during a 6-wk feeding period. Over the first 3-wk period (the incorporation period) the mice were fed a special diet with a high n-3/n-6 PUFA ratio. In the following 3-wk period (the depletion period) the mice were fed a standard chow diet. A linear increase of the concentration of EPA and DHA in erythrocyte membranes was observed during the incorporation period, whereas a stagnation was observed after the second week for leukocytes. The level of EPA did not fall to the background level after the depletion period, and the level of DHA was kept almost constant during the depletion period in the erythrocyte membranes. In leukocytes the concentration of both EPA and DHA decreased during the depletion period, but did not reach the background level after the 3-wk depletion. In conclusion, the kinetics of EPA and DHA in the different cells are different. The rate of incorporation is faster than that of depletion for n-3 PUFA. More n-3 PUFA can be incorporated into leukocytes in comparison with erythrocytes. The ratio of n-3/n 6 PUFA is more important than the amount of n-3 FA in changing the FA compositions of membrane lipids. PMID- 17120928 TI - Partial prevention of hepatic lipid alterations in nude mice by neonatal thymulin gene therapy. AB - During adult life athymic (nude) male mice display not only a severe T-cell related immunodeficiency but also endocrine imbalances and a moderate hyperglycemia. We studied the impact of congenital athymia on hepatic lipid composition and also assessed the ability of neonatal thymulin gene therapy to prevent the effects of athymia. We constructed a recombinant adenoviral vector, RAd-metFTS, expressing a synthetic DNA sequence encoding met-FTS, an analog of the thymic peptide facteur thymique serique (FTS), whose Zn-bound biologically active form is known as thymulin. On postnatal day 1-2 homozygous (nu/nu) nude and heterozygous (nu/+) mice were injected with 10(8) pfu of RAd-metFTS or RAd betagal (control vector) intramuscularly. The animals were processed at 52 d of age. Serum thymulin, glycemia, hepatic phospholipid FA composition and free and esterified cholesterol were determined. Adult homozygous male nudes were significantly (P < 0.01) hyperglycemic when compared with their heterozygous counterparts (2.04 vs. 1.40 g/L, respectively). The relative percentage of 16:0, 18:1 n-9, and 18:1n-7 FA was lower, whereas that of 18:0, 20:4n-6, and 22:6n-3 FA was higher, in hepatic phospholipid (PL) of nu/nu animals as compared with their nu/+ counterparts. Some of these alterations, such as that in the relative content of 22:6n-3 in liver PL and the unsaturation index, were completely or partially prevented by neonatal thymulin gene therapy. We conclude that the thymus influences lipid metabolism and that thymulin is involved in this modulatory activity. PMID- 17120929 TI - Intrasample variability of intramyocellular triacylglycerol. AB - Intrasample variability of intramyocellular triacylglycerol (imcTG) in the skeletal muscle of rats has been examined. Aliquoting after homogenization of muscle samples reduced imcTG variability considerably compared with aliquoting before homogenization. The results suggested that skeletal muscle samples be homogenized before aliquoting in order to reduce imcTG variability. PMID- 17120930 TI - Desaturation indices in liver, muscle, and bone of growing male and female mice fed trans-10,cis-12 conjugated linoleic acid. AB - trans-10,cis-12-CLA (t10,c12-CLA) inhibits lipid deposition in adipose tissue of many species, but it also enhances lipid deposition in liver. We evaluated effects of dietary t10,c12-CLA content and gender on carcass composition, FA profile of selected tissues, and expression of FA synthase (FAS) and stearoyl-CoA desaturase-1 (SCD) mRNA in adipose tissue. Male and female (63 of each) CD-1 mice were assigned a diet containing 0.0, 0.15, or 0.30% t10,c12-CLA at 4 wk of age. Seven mice per dietary group within gender were sacrificed after 2, 4, or 6 wk. The CLA isomer caused dose-dependent reductions in dry carcass weight and fat content, without altering protein content, but carcass fat and epididymal fat pad weights of males were reduced to a greater extent than carcass fat and inguinal fat pad weights of females. FAS and SCD mRNA in adipose tissue was more abundant in females than males, but expression in both genders decreased as the t10,c12 CLA content of the diet increased. Although the weight of gastrocnemius muscle was not influenced by diet, total FA content of the muscle of both genders decreased in response to dietary t10,c12-CLA content. Femur weight of male mice increased as the t10,c12-CLA content of the diet increased, but the weight increase was associated with a reduction in total FA content. The delta 9 desaturation indices for muscle and femur suggested a linear reduction in SCD activity, whereas delta 9 indices for liver indicated linear enhancement of SCD activity. Overall, results suggested that growing male mice were more susceptible than females to t10,c12-CLA inhibition of lipid deposition. PMID- 17120931 TI - Biocatalysis of linoleic acid to conjugated linoleic acid. AB - CLA refers to a group of geometrical and positional isomers of linoleic acid (LA) with conjugated double bonds. CLA has been reported to have diverse health benefits and biological properties. Traditional organic synthesis is highly capital-intensive and results in an isomeric mixture of CLA isomers. Biotechnology presents new alternatives to traditional lipid manufacturing methods. The objective of this study was to examine the effect of protein isolation procedures on linoleate isomerase (LAI) recovery from microbial cells and biocatalysis of LA to CLA. Protein isolation experiments were carried out using Lactobacillus acidophilus L1 and two strains of Lactobacillus reuteri (ATCC 23272 and ATCC 55739). Under the same assay conditions, ATCC 55739 had the highest LAI activity among the microbial cultures examined in this study. Efficiency of cell lysis methods, which included various combinations of lysozyme and mutanolysin treatments in combination with sonication and osmotic rupture of cells with liquid nitrogen, was very low. Although treatment of cell material with a detergent (octylthioglucoyranoside) freed a significant amount of LAI activity into the solution, it was not sufficient to recover all the LAI activity from the residual cells. Crude LAI preparations produced mainly the cis-9,trans 11 CLA isomer. Time and substrate/protein ratio had a significant effect on biocatalysis of LA to CLA. It appears that the mechanism and kinetics of enzymatic conversion of LA to CLA are quite complex and requires further research using pure LAI preparations. PMID- 17120932 TI - Syntheses of conjugated octadecadienoic acids. AB - Three approaches for the synthesis of octadecadienoic acids with conjugated double bond systems are presented: synthesis of (10Z, 12Z)-octadecadienoic acid via an enyne-substructure; the use of an educt with a conjugated double bond system for the synthesis of (10E, 12E)-octadecadienoic acid; and the Suzuki cross coupling for the synthesis of (7E,9Z)-octadecadienoic acid. PMID- 17120933 TI - Alterations of high density lipoprotein subclasses in obese subjects. AB - The object of this study was to investigate the characteristics of lipid metabolism in obese subjects, with particular emphasis on the alteration of HDL subclass contents and distributions. A population of 581 Chinese individuals was divided into four groups (25 underweight subjects, 288 of desirable weight, 187 overweight, and 45 obese) according to body mass index (BMI). Apoprotein A-I (apoA-I) contents of plasma HDL sub-classes were determined by 2-D gel electrophoresis associated with an immunodetection method. The concentrations of TG and the apoA-I content of pre-beta 1-HDL were significantly higher (P < 0.01 and P < 0.01, respectively), but the levels of HDL cholesterol, and the apoA-I contents of HDL2a and HDL2b were significantly lower (P < 0.01, P < 0.05, and P < 0.01, respectively) in obese subjects than in subjects having a desirable weight. Moreover, with the elevation of BMI, small-sized pre-beta 1-HDL increased gradually and significantly, whereas large-sized HDL2b decreased gradually and significantly. Meanwhile, the variations in HDL subclass distribution were more obvious with the elevation of TG levels in obese as well as overweight subjects. In addition, Pearson correlation analysis revealed that BMI and TG levels were positively correlated with pre-beta 1-HDL but negatively correlated with HDL2b. Multiple regression analysis also showed that TG concentrations were associated independently and positively with high pre-beta 1-HDL and independently and negatively with low HDL2b in obese and overweight subjects. The HDL particle size was smaller in obese and overweight subjects. The shift to smaller size was more obvious with the elevation of BMI and TG, especially TG levels. These observations, in turn, indicated that HDL maturation might be abnormal, and reverse cholesterol transport might be impaired. PMID- 17120934 TI - Antioxidant effects of black rice extract through the induction of superoxide dismutase and catalase activities. AB - Our ex vivo study revealed that BRE had significantly stronger ability to inhibit LDL oxidation than white rice extract (WRE). The purpose of this study was to investigate whether black rice extract (BRE) supplementation might ameliorate oxidative stress and enhance antioxidant enzyme activities in HepG2 cells and in C57BL/6 mice. In the cellular study, superoxide anions (O2*-) and reactive oxygen species (ROS) in the BRE group were significantly suppressed. The BRE group also showed significant increases in superoxide dismutase (SOD) and catalase (CAT) activities by 161.6% and 73.4%, respectively. The major components responsible for the free-radical-scavenging and antioxidative properties might be cyanidin-3 O-glucoside chloride and peonidin-3-O-glucuside chloride. In the animal study, male C57BL/6 mice were divided into three groups (control, BRE, and WRE). Plasma HDL-cholesterol was significantly higher, and thiobarbituric, acid-reactive substances were significantly lower in the BRE group, whereas plasma levels of total cholesterol and triglyceride were not affected by BRE supplementation. Increased hepatic SOD and CAT activities were observed in BRE-treated mice as compared to the control mice. However, no changes were detected for the protein expression of antioxidant enzymes by Western blot analysis. Our data suggest that antioxidative effects exerted by BRE are mediated through decreases in free radical generation as well as increases in SOD and CAT activities both in vitro and in vivo. PMID- 17120935 TI - Phospholipid molecular profiles in the seed kernel from different sunflower (Helianthus annuus) mutants. AB - Phospholipids are essential components of plant cell membranes whose acyl composition appears to be influenced by oil composition in the sunflower. In the current study, we have determined the diacylglycerol profile of the main phospholipids using phospholipase C degradation and separation of the diacylglycerols by HPLC and GLC. The main polar lipid molecular species were defined in different classes of sunflower kernel: PC, PE, and PI. The proportions of each were determined at different stages of development in order to define the point at which the mutations carried by each sunflower line affected the phospholipid composition of the seeds. The results indicated that modifications to intraplastidial de novo FA synthesis affected the seed phospholipid profile during the whole period of the seed formation, including accumulation and maturation, whereas the influence of mutations in the endoplasmic reticulum desaturases were more readily detected at later stages of development. These results are discussed in terms of the pathways involved in glycerolipid synthesis and phospholipid conversion in sunflower seeds. PMID- 17120936 TI - Comparative study of lipids in mature seeds of six Cordia species (family Boraginaceae) collected in different regions of Brazil. AB - The oil content, FA, and lipid class composition of the mature seeds of six Cordia species were analyzed. Mature seeds of each species were collected in their natural habitat from 2002 to 2004. The total lipid content varied from 1.9% to 13.2%, there being significant differences between the results found in different years for each species and between the species analyzed. The contents of FFA varied from 2.0% to 7.9% of total lipids. Neutral lipids (NL) were the largest class, making up between 89.6% and 96.4% of the total lipids; the phospholipids (PL) were the second largest class (3.0% to 8.9% of the total lipids), and the glycolipids (GL) were the smallest class (0.6 to 3.4%). The presence of GLA was determined in each class of lipids; it is predominant in the NL. Levels of GLA ranged from 1.2% to 6.8% of total seed FA. This is, to our knowledge the first study of lipid composition in seeds of species of Cordia from Brazil. PMID- 17120937 TI - A three-decade perspective on anesthesia safety. AB - Modern medical practice, and particularly that within the hospital environment, has been under intense scrutiny in an attempt to improve patient safety and optimize outcomes. Anesthesiology has been cited as among the most successful specialties effecting improvements. According to the Institute of Medicine's 1999 report, To Err is Human, "... anesthesiology has successfully reduced anesthesia mortality rates from two deaths per 10,000 anesthetics administered, to one death per 200,000 to 300,000 anesthetics administered." The current report reviews representative highlights from 30 years of progress in improving anesthesiology safety and offers a speculative synthesis of the factors critical to past and future successes. The seven identified points include 1) the emergence of a champion and his allies, 2) initial efforts to identify and quantify broad reaching problems, 3) research addressing intellectually "amusing" problems of relevance to practitioners, 4) reaching out to others with focused expertise in problem prevention and problem solving, 5) sharing the responsibility for quality and safety improvement with other specialties, 6) expanding buy-in and participation within the anesthesia community, and 7) preparing for the future. The factors provide not only an accounting of anesthesiologists' successes, but also a road map for other groups and specialties desiring to emulate the anesthesiologists' experience. PMID- 17120938 TI - Early efforts with quality at centers for Medicare and Medicaid services and judicious use of its powers as a payor. PMID- 17120939 TI - Safety, quality, and the National Surgical Quality Improvement Program. AB - The Institute of Medicine 1999 publication, To Err is Human, focused attention on preventable provider errors in surgery, and prompted numerous new national initiatives to improve patient safety. It is uncertain whether these initiatives have actually improved patient safety, mainly because of the lack of a quantitative metric for the assessment of patient safety in surgery. A 15-year experience with the National Surgical Quality Improvement Program, which originated in the Veteran's Administration in 1991 and was recently made available to the private sector, prompts the surgical community to place patient safety in surgery within a much larger conceptual framework than that of the Institute of Medicine report, and provides a quantitative metric for the assessment of patient safety initiatives. This conceptual framework defines patient safety in surgery as safety from all adverse outcomes (not only preventable errors and sentinel events); regards safety as an integral part of quality of surgical care; recognizes that adverse outcomes, and hence patient safety, are primarily determined by quality of systems of care; and uses comparative risk-adjusted outcome data as a metric for the identification of system problems and for the assessment and improvement of patient safety from adverse outcomes. PMID- 17120940 TI - Surgical quality programs in the Veterans Health Administration. AB - This review describes the development, implementation, and current status of programs that promote and maintain surgical quality performance within the Department of Veterans Affairs, Veterans Health Administration (VHA). It also considers evolving initiatives to improve surgical outcomes and enhance patient safety. The VHA is a nationwide health care system operated and funded by the federal government. It consists of 154 federal hospitals and 875 clinics staffed by 16,000 nurses, 33,000 other health care professionals, 15,000 doctors, and 25,000 university-affiliated faculty physicians. The VHA trains 90,000 health care professionals, and additionally its activities are supported by 140,000 dedicated volunteers. More than 1000 surgeons in all specialties serve in its facilities. Since these programs began more than 19 years ago, there has been a steady decrease in both risk-adjusted and actual mortality rates tracked annually. PMID- 17120941 TI - American College of Surgeons remains committed to patient safety. AB - Since 1913 the American College of Surgeons has addressed patient safety as a top priority, so they are pleased to contribute this article offering the College's perspective on this critical subject. More specifically, this piece reviews the College's perennial efforts to ensure surgeons and hospitals access to scientifically verifiable standards, availability of effective quality improvement tools, and a better understanding of errors in care. Additionally, they examine the cultural changes required within surgery and provide an overview of the College's recent initiatives in research, accreditation, and education. PMID- 17120942 TI - The Surgical Infection Prevention and Surgical Care Improvement Projects: promises and pitfalls. AB - Variations in outcomes for patients who have surgery are well known, and there is extensive evidence that failure to apply standards of care known to prevent adverse events results in patient harm. Infections and postoperative sepsis, cardiovascular complications, respiratory complications, and thromboembolic complications represent some of the most common adverse events that occur after surgery. Patients who experience postoperative complications have increased hospital length of stay, readmission rates, and mortality rates; in addition, costs of care are increased for patients, hospitals, and payers. In 2002, the Centers for Medicare and Medicaid Services, in collaboration with the Centers for Disease Control and Prevention, implemented the Surgical Infection Prevention Project to decrease the morbidity and mortality associated with postoperative surgical site infections. More recently, the Surgical Care Improvement Project, a national quality partnership of organizations committed to improving the safety of surgical care has been implemented. Although the Surgical Care Project does not focus on the complete set of important surgical quality issues, it does provide the incentive and infrastructure for national data collection and quality improvement activities for hospitals. There is now a strong national commitment to measure processes and outcomes of care for surgery in the United States. PMID- 17120943 TI - The evolution of guidelines toward standards of practice. AB - The development of practice guidelines and its many synonyms has been the source of concern and a large amount of work in surgical specialty practice, especially over the last decade. These guidelines represent improvements and refinements. If they are strongly adhered to, standards of practice without consideration of outliers, physician judgment, and the man on the scene, then they have the capacity for great misunderstanding and harm. In general, I firmly believe that surgeons, their hospitals, and everybody involved in the process will find the preparation for transparency a positive experience, and that these guidelines, when properly assessed and refined by practicing surgical specialists, can become a strong asset not only for ourselves and our profession but also for our patients. In the future, guidelines must be carefully stated with specific exceptions and caveats. These guidelines need the respect, the careful judgment of the physician at the time and place, and the relative roles of a consultation and even second opinions. We must also be aware of the substantial capacity for influence of these standards by industry. A careful and ongoing analysis of the self-serving aspects of such work is essential. However, these observations and our work make the decision for elective surgical operations safer than it has ever been. PMID- 17120944 TI - The hazards of using administrative data to measure surgical quality. AB - Administrative claims data have been used to measure risk-adjusted clinical outcomes of hospitalized patients. These data have been criticized because they cannot differentiate risk factors present at the time of admission from complications that occur during hospitalization. This paper illustrates how valid risk-adjustment can be achieved by enhancing administrative data with a present on-admission code, admission laboratory data, and admission vital signs. Examples are presented for inpatient mortality rates following craniotomy and rates of postoperative sepsis after elective surgical procedures. Administrative claims data alone yielded a risk-adjustment model with 10 variables and a C-statistic of 0.891 for mortality after craniotomy, and a model with 18 variables and a C statistic of 0.827 for postoperative sepsis. In contrast, the combination of administrative data and clinical data abstracted from medical records increased the number of variables in the craniotomy model to 21 with a C-statistic of 0.923, and the number of variables in the postoperative sepsis model to 29 with a C-statistic of 0.858. Use of only administrative data resulted in unacceptable amounts of systematic bias in 24 per cent of hospitals for craniotomy and 19 per cent of hospitals for postoperative sepsis. Addition of a present-on-admission code, laboratory data, and vital signs reduced the percentage of hospitals with unacceptable bias to two percent both for craniotomy and for postoperative sepsis. These illustrations demonstrate suboptimal risk stratification with administrative claims data only, but show that present-on-admission coding combined with readily available laboratory data and vital signs can support accurate risk-adjustment for the assessment of surgical outcomes. PMID- 17120945 TI - The volume-outcome debate revisited. AB - Multiple studies support the intuitive association between higher provider procedure volume and better clinical outcomes. Health care purchasers and payers have been seeking ways to direct patients to high-volume providers to improve the quality of care received and to avoid costs associated with higher surgical morbidity. Volume-based referral has faced resistance from providers who are concerned that the use of volume instead of more direct measures of surgical quality will result in unfair discrimination. On close examination, volume-based referral policies also appear to be more congruent with payers' interests than the interests of individual patients and providers. Furthermore, a policy of volume-based referral does not address surgical quality directly, is applicable to only a very small segment of surgical care, and is logistically problematic. However, in the absence of viable alternative measures of surgical quality, imperfect proxies such as volume will likely continue to be a significant part of the national dialogue surrounding surgical quality. PMID- 17120946 TI - The challenge of small numbers to the contemporary excellence of elective specialty surgery. PMID- 17120947 TI - The holy Grail of surgical quality Improvement: process measures or risk-adjusted outcomes? AB - Quality of care in surgery has garnered increased attention. Focus on the structure, processes of care, and use of risk-adjusted outcomes has offered different possible solutions. Volume-outcome studies have consistently demonstrated relationships between higher surgeon and hospital volume and favorable outcomes. The policy implications for quality improvement remain unclear. Recent efforts have focused on the use of risk-adjusted outcomes, such as the American College of Surgeons National Surgical Quality Improvement Program, to drive quality improvement. Conversely, large efforts, mandated by the Center for Medicare and Medicaid Services, have focused on process measures, such as perioperative complication prevention. For the future of surgical quality improvement, the combination of process measures and risk-adjusted outcomes are essential. It is only through the combined use of improved understanding of the relationship of processes of care and outcomes that we will make surgical care safer and improve quality. PMID- 17120948 TI - Quality and safety performance in teaching hospitals. AB - The Hospital Quality Alliance created a vehicle to display Hospital Performance data which is known as Hospital Compare. Overall, the data shows that teaching hospitals perform very well in the areas of Heart Failure and Heart Attack and not as well in Pneumonia care. Unique issues at teaching hospitals, such as timing for specific patient services, continue to be a concern in achieving high scores relative to their non-teaching peers. Most hospitals and specifically surgical services will be challenged in the upcoming years with the addition of the Surgical Care Improvement Project (SCIP) measures as we move into the pay-for performance era. PMID- 17120949 TI - Does outcomes research impact quality? Examples from bariatric surgery. AB - This manuscript addresses the question "Does outcomes research affect quality?" using examples from the field of bariatric surgery. The roles that outcomes research has played in each of the four major recent events in bariatric surgery are examined. In the first three major events, which include 1) the National Institutes of Health Consensus Conference on Bariatric Surgery in 1991, 2) the dramatic increase in numbers of bariatric operations performed, and 3) the move toward a laparoscopic approach in bariatric surgery, a multitude of outcomes studies seem to be the result, but not the cause, of these changes in the field of bariatric surgery. However, for the most recent event, the 2006 Centers for Medicare and Medicaid Services National Coverage Determination for bariatric surgery and the introduction of accreditation in general surgery, outcomes research has played a significant role in the determination of policy and, ultimately, quality. PMID- 17120950 TI - An eight-year analysis of surgical morbidity and mortality: data and solutions. AB - In this article, a reproducible process for presenting, analyzing, and reducing early and late surgical morbidity and mortality (M and M) is detailed. All M and M cases presented from 1998 through 2005 at Monmouth Medical Center were categorized. Residents and nurses were empowered to report the complications. The five major categories were overwhelming disease on admission, delays in treatment, diagnostic or judgment complications, treatment complications, and technical complications. From the 53,541 operations performed over 8 years, 714 patients were presented, which included 147 deaths and 1,132 category entries. The most common problems were technical complications in 474 (66.4%) patients. The data have generated actionable solutions, many with low barriers to adoption, resulting in safer, less expensive surgical management. Surgical outcome benchmarks have been established and are used for credentialing surgeons. The "Hostile Abdomen Index" has been developed to assess the safest choice for abdominal operative access, pre- and intraoperatively. We explained the real-time process that generated solutions for the entire department as well as changes relevant to residency training and individual operative techniques. PMID- 17120951 TI - The challenge of changing roles and improving surgical care now: Crew Resource Management approach. AB - Many surgeons are also pilots; the two activities demand similar skill sets. Surgeons have developed an interest in aviation models for managing risk and reducing adverse events, such as Crew Resource Management training. This article provides seven suggestions from aviators that might be adopted by surgeons in an effort to improve surgical care and mitigate patient harm. Each suggestion is offered based on the value added to aviation, with an acknowledgment that the suggestion may be more or less applicable in surgery. The suggestions for dealing with the changing roles for surgeons are: Crew Resource Management-type training to improve teamwork should be required for hospital credentialing, surgeons should brief the operating room team before an operation, surgeons should write standards specific to their organization, surgeons should recognize fatigue and age as factors in performance, surgeons should have "check-rides" as a part of the credentialing process, surgeons should abandon the mortality and morbidity conference in favor of a data collection system that effectively examines adverse events for root causes of error, and all members of the surgical team should be subject to mandatory, random drug testing. PMID- 17120952 TI - How a system for reporting medical errors can and cannot improve patient safety. AB - The Institute of Medicine has recommended systems for reporting medical errors. This article discusses the necessary components of patient safety databases, steps for implementing patient safety reporting systems, what systems can do, what they cannot do, and motivations for physician participation. An ideal system captures adverse events, when care harms patients, and near misses, when errors occur without any harm. Near misses signal system weaknesses and, because harm did not occur, may provide insight into solutions. With an integrated system, medical errors can be linked to patient and team characteristics. Confidentiality and ease of use are important incentives in reporting. Confidentiality is preferred to anonymity to allow follow-up. Analysis and feedback are critical. Reporting systems need to be linked to organizational leaders who can act on the conclusions of reports. The use of statistics is limited by the absence of reliable numerators and denominators. Solutions should focus on changing the cultural environment. Patient safety reporting systems can help bring to light, monitor, and correct systems of care that produces medical errors. They are useful components of the patient safety and quality improvement initiatives of healthcare systems and they warrant involvement by physicians. PMID- 17120953 TI - Maintenance of Certification: American Board of Surgery goals. PMID- 17120954 TI - Simulation: a strategy for success in quality and safety in pay-for-performance environments. AB - Pay-for-Performance appears to be another step in our ever-changing healthcare environment. In most of the white papers, reports, and web pages devoted to improving the quality of healthcare in America, there is a failure to recognize or list medical simulation as a methodology to reduce the costs of implementation and to speed transition to the new order. The Agency for Healthcare Research and Quality is funding research in simulation to improve quality. This article outlines the rationales for using simulation and how simulation can benefit all involved. With a paucity of proof that simulation can deliver in terms of improving the quality of healthcare, the mass of evidence has been from observation and anecdotal tales of medical professionals that simulation is a valid tool. This article correlates the use of simulation in other nonmedical pay for-performance professions to similar situations in medicine as some other evidence that simulation should be considered a viable option. I conclude by relating the individual strengths of simulation to the six quality initiatives of the Institute of Medicine's second report from the Committee on Quality of Health Care in America. Simulation can work to enhance the assimilation of change with each of these initiatives and help to reduce the costs of doing so. There are limitations to simulation, but used within those limitations, simulation should prove to be a powerful tool. PMID- 17120955 TI - Making the operating room of the future safer. AB - There is an increasing demand for interventions to improve patient safety, but there is limited data to guide such reform. In particular, because much of the existing research is outcome-driven, we have a limited understanding of the factors and process variations that influence safety in the operating room. In this article, we start with an overview of safety terminology, suggesting a model that emphasizes "safety" rather than "error" and that can encompass the spectrum of events occurring in the operating room. Next, we provide an introduction to techniques that can be used to understand safety at the point of care and we review the data that exists relating such studies to improved outcomes. Future work in this area will need to prospectively study the processes and factors that impact patient safety and vulnerability in the operating room. PMID- 17120956 TI - Morbidity and mortality in vascular surgery: the Kentucky experience with a statewide database. AB - Efforts at improvement in quality of care for surgical patients have required multiple strategies that include local, regional, and national efforts to influence processes and outcomes, and examination of outcome databases with and without risk stratification. In the mid to late 1980s, there was an effort to examine the outcome of two high-risk procedures (carotid endarterectomy and abdominal aortic aneurysm repair) on all Medicare patients in the state of Kentucky with an effort to determine the outcome of their treatment and improve the quality of care delivered to them. Even though this experience is somewhat dated, it still represents a unique examination of a large number of patients treated by numerous surgeons. Furthermore, all charts reviewed by the author presented an opportunity to compare actual patient data with that obtained from an administrative data set. This report also examines the author's attempts at improving outcome. PMID- 17120957 TI - The practicing surgeon as a member of the quality-safety team. AB - The practicing surgeon is a valuable member of the quality-safety team and is often an underutilized data source for quality initiatives. The authors describe how their efforts in Kentucky, during a 10-year period, encouraged surgeons to become leaders in the quality initiative. Their experience began with the establishment of an organization of surgeons devoted to quality health care and cost control. As their efforts expanded and they gained experience, they were well prepared to transition to a regional and national quality initiative as part of a collaborative effort with the Centers for Medicare and Medicaid Services in the 2004 Surgical Care Improvement Project pilot. As a result of this ongoing experience, the authors we have been able to affect the quality of health care and have a positive influence on health care cost. They have demonstrated that surgeons will participate in and lead quality initiatives, and that these efforts foster an environment of cooperation between surgeons and hospitals. PMID- 17120958 TI - Linking processes and outcomes to improve surgical performance: a new approach to morbidity and mortality peer review. AB - Peer review of surgical cases resulting in death or potentially avoidable complications is a longstanding tradition, but intensive reviews of individual cases rarely produces tangible improvements in clinical outcomes. The systematic comparison of care received by patients who experienced adverse outcomes to care received by patients who had uneventful surgical courses is a promising alternative to intensive case review. However, the results of these studies may be misleading because physicians often choose interventions based on their perceptions of patients' preoperative risks, and higher adverse outcome rates among patients with higher preoperative risk may distort comparisons of alternative interventions. The creation of a control sample by carefully matching each patient who experienced an adverse outcome to a patient who had a similar preoperative risk but did not experience an adverse outcome can overcome this problem and provide excellent insight into how to improve clinical performance most effectively and efficiently. By using currently available electronic data to compute each patient's risk of an adverse outcome, a series of cases with adverse outcomes can be matched to an equal number of controls. Peer review committees can then direct focused data collection and analyses of potentially critical processes of care to determine which, if any, are associated with significantly poorer clinical results. A simulated scenario is analyzed in detail to illustrate the ability of this technique to correctly determine best practices when other approaches fail to do so. PMID- 17120959 TI - Where is the captain of the ship when you need her? PMID- 17120960 TI - Efficacy of ivermectin treatment of cutaneous gnathostomiasis evaluated by placebo-controlled trial. AB - Previous studies have revealed that ivermectin treatment for gnathostomiasis can reduce parasitic loads in animals and make recurrent subcutaneous swelling subside in 76% of patients. Our study aimed to evaluate the efficacy of ivermectin for cutaneous gnathostomiasis treatment in a placebo-controlled trial. This study was a prospective randomized placebo-controlled study performed at The Bangkok Hospital for Tropical Diseases, Mahidol University, Thailand. Thirty patients with a serologically confirmed diagnosis of cutaneous gnathostomiasis were enrolled. Seventeen patients in the ivermectin treated group received a single dose of 12 mg ivermectin (200 microg/kg bodyweight), while 13 patients in the control group received a single dose of 40 mg of vitamin B1. The follow-up period was 1 year. Of the 17 patients, 7 (41.2%) responded to ivermectin, while no patient responded to placebo. The mean (95% Cl) time to the first recurrence of subcutaneous swelling with ivermectin and in the placebo groups were 257 (184 331) and 146 (42-250) days, respectively, (p=0.102). Although this study revealed no significant difference in the mean time to first recurrence of swelling between the ivermectin and placebo groups, there was a trend towards ivermectin efficacy against gnathostomiasis in previous animal and human studies. Further studies with different doses of ivermectin and larger sample sizes, and close monitoring for ivermectin tolerability and treatment response are necessary to confirm an efficacy of ivermectin. PMID- 17120961 TI - Thiamine deficiency and parasitic infection in rural Thai children. AB - A study was carried out to determine whether thiamine deficiency in northeast Thailand is linked with parasitic infection. We assessed the thiamine status and tested for parasitic infections in 231 children between 6 and 12 years of age in Nam Phong District of Khon Kaen Province, Thailand. Thiamine deficiency [thiamine pyrophosphate effect (TPPE) > 20%] and parasitic infections (mostly Strongyloides stercoralis) were identified in 16 and 45% of the children, respectively. No association between thiamine deficiency and parasitic infection was found. The general health of the children, according to Thai standard anthropometric curves, was within the normal range. The suboptimal thiamine status and the high prevalence of parasitic infection require attention. PMID- 17120962 TI - Mucosal mast cell responses in rats (Rattus norvegicus) experimentally infected with Centrocestus caninus. AB - Mucosal mast cell (MMC) responses and worm recovery rates in rats experimentally infected with Centrocestus caninus were investigated. Metacercariae of C. caninus, procured from goldfish, Carassius auratus, were orally administered to twenty-five male rats (300 metacercariae each rat). The infected rats were sacrificed on days 3, 7, 14, 21 and 28 post-infection (PI) along with the control rats. Worm recovery was performed from each part of small intestine. To investigate MMC, duodenal, jejunal and ileal paraffinized-tissue sections were processed and stained with 1% alcian blue and 0.5% safranin-O. The average worm recovery rates were 42.8, 37.7, 21.2, 12.5 and 3.7% on days 3, 7, 14, 21 and 28 PI, respectively. The majority of the worms (98.9%) were collected from the duodenum and jejunum. The MMC numbers in the infected rats were significantly higher than those of the controls (p<0.05). A peak level was observed on days 14 PI and the numbers gradually decreased thereafter. The results reveal that MMC plays an important role in the expulsion of C. caninus from the host intestine. A more precise description of the role the MMC plays in helminth expulsion is still needed to understand the mechanism of host defense against intestinal helminthic infection, along with other effector cells, such as goblet cells. PMID- 17120963 TI - An effective indirect fluorescent antibody test for diagnosis of intestinal acariasis. AB - Adult mites' bodies of Dermatophagoides farinae were used as antigen in an indirect fluorescent antibody test (IFAT) to detect mite-specific IgG in sera of 48 patients with intestinal acariasis based on stool examination. Antibody titers with positive reaction ranged from 1:4 to 1:512 in 48 patients with intestinal acariasis. If antibody titers > or = 1:16 is regarded as being positive, the positive rate of patients detected with IFAT was 92%. PMID- 17120964 TI - Longevity of Aedes aegypti (Diptera: Culicidae) compared in cages and field under ambient conditions in rural Thailand. AB - Aedes aegypti (L.) were exposed to all of the physiological stresses of a natural environment, without mortality from predation or from the defensive behavior of bitten hosts. Each replicate consisted of four cages containing 50 male and 50 female, locally reared Ae. aegypti. The cages were placed in the bedroom and kitchen of a typical Thai house in the village of Hua Samrong, Chachoengsao Province. Replicates were repeated five times between October 1991 and June 1992. Mosquitoes had constant access to sugar and were offered blood meals every day. The number of dead mosquitoes and eggs were recorded daily for 30 days. Indoor maximum temperatures were high throughout the year, ranging from a mean of 32.8 degrees C in October/November to 37.6 degrees C in March-April, with an absolute maximum of 41 degrees C. Survival in cages was related to temperature, with the force of mortality lowest in November-December (0.002) and highest in May-June (0.043). The negative slope of cohort survival was also greatest in the warmest months. Egg laying rate was lower in the cooler months (minimum 16.4 eggs/female/day in November-December), but did not vary greatly in absolute value (maximum 22.7 eggs/female/day in March-April). Statistically, survival of females in cages was much greater than survival calculated from mark-release-recapture studies conducted by other authors in Hua Samrong. The difference in survival for mosquitoes released in the field and those confined to cages suggests that predation or defensive behavior may be important in regulating adult populations of this vector. PMID- 17120965 TI - The relationship between the abundance of Mansonia mosquitoes inhabiting a peat swamp forest and remotely sensed data. AB - The present study aimed to demonstrate the relationship of some environmental factors, vegetation greenness index (NDVI) and land surface temperature (LST), with the seasonal variations of Mansonia bonneae and Ma. uniformis in Khosit Subdistrict, Narathiwat Province. It was found that the Mansonia population lagged one month behind but correlated positively to NDVI, LST and rainfall. A rise in the number of mosquitoes was directly related to a rise in vegetation, temperature and rainfall. PMID- 17120966 TI - Double infection of heteroserotypes of dengue viruses in field populations of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) and serological features of dengue viruses found in patients in southern Thailand. AB - In order to understand more about the epidemiology of DHF, a study of the type of dengue viruses and vectors under natural conditions was carried out. Mosquito vectors in the field and the serum of DHF patients in southern Thailand were examined. The two mosquito species are abundant and DHF incidence remains high in this region. Dengue viruses were examined in field-caught mosquitoes by RT-PCR technique. The mosquitoes were caught in 4 provinces: Krabi, Phuket, Phang-Nga and Surat Thani during the late dry season until the early rainy season in 2005. Three dengue serotypes (DEN-2, DEN-3, DEN-4) were detected in Ae. aegypti males and females, and 2 (DEN-2, DEN-3) were detected in Ae. albopictus females. Double infection with 2 serotypes of dengue viruses (DEN-2 and DEN-3) were detected in Ae. aegypti males and females and Ae. albopictus females. DEN-2 and DEN-1 were the most prevalent serotypes found in the serum of the patients in this area, followed by DEN-4 and DEN-3. The prevalence of the predominant dengue serotype varied from province to province. Detection of viruses in adult male mosquitoes reveals the role of transovarial transmission of dengue viruses in field populations of DHF vectors and elucidates circulation of dengue viruses in vectors in the natural environment of endemic areas. The incidence of multiple serotypes of dengue virus in Ae. aegypti and Ae. albopictus in the same area points toward a high risk for an epidemic of DHF. These findings provide greater understanding of the relationship among mosquito vectors, virus transmission and DHF epidemiology in endemic areas. PMID- 17120967 TI - Quantitative detection of human immunodeficiency virus type 1 (HIV-1) viral load by real-time RT-PCR assay using self-quenched fluorogenic primers. AB - HIV-1 viral load is a basic marker to evaluate the severity of HIV-1 related diseases and to monitor the effectiveness of treatment. A method based on real time RT-PCR technology has been developed to quantify HIV-1 RNA using self quenched fluorogenic primers known as LUX primers. They were used in this study to recognize a low variable gag region of subtype E and B consensus sequences. Specificity was verified by amplicon melting temperatures. An external standard curve was constructed with 10 fold serial dilutions of synthetic HIV-gag RNA. A broad range linear relationship (10 to 10(6) copies/ml) was observed between the number of PCR cycles needed to detect a fluorescent signal and the number of RNA copies. The intra- and inter-assay coefficients of variation were 0.72 to 2.54% and 3.14 to 8.83%, respectively, thus indicating good reproducibility. Thirty out of fifty HIV-infected individual plasma samples were quantified by this method and compared with the AMPLICOR HIV-1 Monitor assay, which is widely considered the reference technique for HIV-RNA viral load measurement. The results indicate that the AMPLICOR HIV-1 Monitor assay and real-time RT-PCR using LUX primers are in good agreement (mean difference in log10 copies/ml+/-2 standard deviations = 0.21+/-1.34). PMID- 17120968 TI - A household survey to assess the burden of influenza in rural Thailand. AB - Little is known about the disease burden of influenza in middle-income tropical countries like Thailand. The recent outbreak of avian influenza (H5N1) and studies on influenza from neighboring countries highlight the need for data on incidence, access to care, and health care cost. In May/ June 2003, we conducted a province-wide household survey using two-stage cluster sampling to determine the burden of influenza-like illness in Sa Kaeo Province. We used the total number of reported influenza that occurred in May 2003 and a prospective study of outpatient influenza in clinic patients to develop an estimate of the annualized incidence of influenza. Of 718 subjects, 16 (2.2%) suffered an episode of influenza-like illness in the preceding month; 14 sought care, of whom 7 went to a hospital facility. Fifty percent reported missing on average 3 days of work or school. The total individual cost per illness episode was 663 baht (15.78 US dollars). The proportion of outpatients with influenza-like illness caused by an influenza virus in May was 16% and the annualized influenza incidence was estimated to be 5,941/100,000 in Sa Kaeo Province. This survey adds to information indicating that in rural Thailand, the burden of influenza is substantial and costs associated with an illness episode are up to 20% of an average monthly income. PMID- 17120969 TI - One-tube multiplex PCR method for rapid identification of mycobacterium tuberculosis. AB - A rapid, inexpensive, simple, and accurate multiplex polymerase chain reaction (PCR) was developed in a single tube for identification of Mycobacterium tuberculosis. Assessment of sensitivity and specificity of simple PCR was performed with 116 strains of M. tuberculosis complex (MTC) and 144 strains of nontuberculous mycobacteria (NTM) compared with the biochemical method. Specific amplification of KS4, MTC-specific DNA fragment, was found in 98% (114/116) of MTC and not detected in 99% (143/144) of NTM. Amplification of the mtp40 gene revealed 95% sensitivity (100/105 strains of M. tuberculosis) and 77% specificity (not found in 119/155 mycobacterial strains). A multiplex PCR method based on the combination of KS4- and mtp40-derived primers was used for identification of M. tuberculosis. Crude DNA from slow growing mycobacteria with cream rough colonies that showed both 768-bp amplified product for KS4 and 396-bp for mtp40 was identified as M. tuberculosis whereas that from MTC gave only the 768-bp product. PMID- 17120970 TI - Development of a combined air sampling and quantitative real-time PCR method for detection of Legionella spp. AB - The objective of this study was to develop and optimize the combined methods of air sampling and real time polymerase chain reaction (real-time PCR) for quantifying aerosol Legionella spp. Primers and TaqMan hydrolysis probe based on 5S rRNA gene specific for Legionella spp were used to amplify a specific DNA product of 84 bp. The impinger air sampler plus T-100 sampling pump was used to collect aerosol Legionella and as low as 10 fg of Legionella DNA per reaction could detected. Preliminary studies demonstrated that the developed method could detect aerosol Legionella spp 1.5-185 organisms /500 l of air within 5 hours, in contrast to culture method, that required a minimum of 7-10 days. PMID- 17120971 TI - Evaluation of an algorithm for persistent/ chronic diarrhea in children at a community hospital adjoining slums in Agra, north India. AB - The objective of this study was to evaluate an algorithm for the management of children with persistent/chronic diarrhea at a community level hospital. The study was carried out in the pediatric OPD of a 150 bed trust hospital catering to children from poor, rural and urban slums. Fifty clinically stable children (6 months-5 years old, mean = 19.7 months) with persistent or chronic diarrhea refusing admission, being managed on an outpatient basis, were enrolled prospectively. A detailed history and physical examination were done for each child to ascertain the cause of diarrhea. They were managed using a pre-tested simplified algorithm and monitored for symptom improvement using a questionnaire 15 days, 1 month and 3 months after initiation of therapy. The average cost for treatment of each child was also calculated. Twenty-one (42%) children had persistent diarrhea. Seven (14%) infants with a typical history of lactose malabsorption responded to a trial of WHO feeding protocols or lactose/sucrose free milk. Four (8%) infants had chronic non-specific diarrhea. A total of 71.8% (28/39) of children were treated satisfactorily with albendazole or metronidazole and Cotrimaxazole along with hematinics and multivitamins. Three (6%) children were diagnosed with abdominal tuberculosis. Four (8%) had raised anti-tissue tranglutaminase antibodies (age 18-34 months). The algorithm used was successful in managing all the children with chronic diarrhea. The average cost per managed case was US$10. Further, multi-center evaluations of similar algorithms are needed to validate the observations in the present study. PMID- 17120972 TI - The feasibility of a school-based VI polysaccharide vaccine mass immunization campaign in Hue City, central Vietnam: streamlining a typhoid fever preventive strategy. AB - We report the coverage, safety, and logistics of a school-based typhoid fever immunization campaign that took place in Hue City, central Vietnam; a typhoid fever endemic area. A cluster-randomized evaluation-blinded controlled trial was designed where 68 schools (cluster) were randomly allocated the single dose Vi polysaccharide vaccine (Typherix) or the active control hepatitis A vaccine (Havrix). A safety surveillance system was implemented. A total of 32,267 children were immunized with a coverage of 57.5%. Strong predictors for vaccination were attending primary schools, peri-urban location of the school, and low family income. Human resources were mainly schoolteachers and the campaign was completed in about 1 month. Most adverse events reported were mild. Safe injection and safe sharp-waste disposal practices were followed. A typhoid fever school-based immunization campaign was safe and logistically possible. Coverage was moderate and can be interpreted as the minimum that could have been achievable because individual written informed consent procedures were sought for the first time in Hue City and the trial nature of the campaign. The lessons learned, together with cost-effectiveness results to be obtained by the end of follow-up period, will hopefully accelerate the introduction of Vi typhoid fever vaccine in Vietnam. PMID- 17120973 TI - Effects of sonicated Prevotella intermedia, Fusobacterium nucleatum and Lactobacillus casei extracts on interleukin-8 production by human dental pulp cells. AB - The objective of this study was to determine the effects of Prevotella intermedia, Fusobacterium nucleatum and Lactobacillus casei on the production of IL-8 by human dental pulp cells. Human dental pulp cells from teeth of young patients (aged 18-25 years) were cultured and tested with sonicated P. intermedia ATCC 25611, F. nucleatum ATCC 25586 and L. casei ATCC 4646 extracts. IL-8 secreted into the culture supernatants were measured at 6, 12 and 24 hours using a quantitative sandwich enzyme immunoassay technique. Cell viability was evaluated using trypan blue exclusion technique. IL-8 production by human dental pulp cells increased significantly at 12 and 24 hours after exposure to P. intermedia and F. nucleatum, whereas L. casei extract exhibited low IL-8 production. The sonicated bacterial extracts did not significantly affect viability or total number of dental pulp cells. PMID- 17120974 TI - Effect of Cha-em Thai mouthwash on salivary levels of mutans streptococci and total IgA. AB - The aim of this study was to investigate the effect of Cha-em Thai (Albizia myriophylla) mouthwash on the mutans streptococci (MS) and immunoglobulin A (IgA) level in saliva. Sixty-seven schoolchildren, age 6-12 years, with MS more than 1x10(5) cfu per milliliter of saliva were entered in this study. They were divided into two balanced groups according to their baseline MS counts (> or = 10(5) cfu/ml). Each group was randomly assigned to use either the Cha-em Thai mouthwash or the placebo mouthwash. After twice daily rinses with their mouthwash for 2 weeks, stimulated saliva were collected and analysed. The MS counts and IgA levels for the two groups were compared. Those rinsing with Cha-em Thai mouthwash showed a significant reduction in MS counts (p<0.05), but the IgA levels were not different. Those rinsing with the placebo mouthwash showed no statistically significant differences in the MS counts or the IgA levels (p>0.05). The results indicate that twice daily use of Cha-em Thai mouthwash can reduce the levels of MS in saliva. PMID- 17120975 TI - Anti-head lice effect of Annona squamosa seeds. AB - The present study focused on the separation and identification of the active compounds against head lice from the hexane extract of Annona squamosa L seed. Chromatographic and spectroscopic techniques revealed that two major compounds of the hexane seed extract were oleic acid and triglyceride with one oleate ester. The yields of these compounds were 13.25% and 7.74% dry weight, respectively. The compounds were tested in vitro against head lice, comparing to the crude hexane extract of the seed. The triglyceride with one oleate ester and the crude hexane extract diluted with coconut oil 1:1. These compounds were found to kill all tested head lice in 49, 11 and 30 minutes, respectively. The triglyceride ester can be used as a marker for quantitative analysis of the active compound for quality control of the raw material A. squamosa seed and its extract. This first finding will be useful for quality assessment and the chemical stability of the antihead lice preparation from this plant. PMID- 17120976 TI - Prevalence and associated factors of head lice infestation among primary schoolchildren in Kelantan, Malaysia. AB - Head lice infestation contributes a significant morbidity among schoolchildren in Malaysia. A cross-sectional study was designed to determine the prevalence and associated factors of head lice infestation among primary schoolchildren in Kelantan, Malaysia. Six schools were randomly selected from three sub-districts of Kuala Krai, Kelantan. A total of 463 eleven-year-old pupils were screened by visual scalp examination and fine-toothed combing. Self-administered questionnaire was used to collect data on socio-demography and associated factors of head lice infestation. The prevalence of head lice infestation was 35.0% (95% Cl: 30.6, 39.3) with 11.9% inactive, 23.1% active, 18.2% light and 16.8% heavy infestations. The associated factors were girls; family income of RM247 or less; head lice infestation of family member and having four or more siblings. The high prevalence of head lice infestation in this study indicates the need for regular school health program that emphasis on the eradication of head lice. The significant associated factors identified in this study reconfirm the importance of controlling the transmissibility of head lice. Pupils and parents should be informed regarding factors that may facilitate the transmission of head lice. PMID- 17120977 TI - Preparation of in vivo cow control blood samples for cadmium analysis. AB - Quality control is essential for any analysis in the laboratory. The objective of this study was to prepare in vivo cow control blood samples. The experiment was performed by feeding cows with a single dose of cadmium in the form of cadmium chloride, withdrawing the blood at an appropriate time to get the highest level of cadmium and detecting the level of cadmium in the blood. It was found that feeding the cow a single dose of 0.06 mg cadmium per kg body weight resulted in the highest cadmium level of 3.622 microg/l 30-60 minutes after feeding. The samples were homogeneous because feeding the cows with single dose of cadmium let the cadmium be absorbed and distributed naturally. In addition, the samples were stable during transport. Therefore, they may be used as quality control samples to detect cadmium levels without using a lyophilized process. They could be used for proficiency testing and to evaluate whole blood analysis in the laboratory. PMID- 17120978 TI - Thalassemia among blood donors at the Hospital Universiti Sains Malaysia. AB - The aim of this study was to screen and identify the types of thalassemia among blood donors at the Hospital Universiti Sains Malaysia (HUSM). Thalassemia screening was performed by hemoglobin electrophoresis. A total number of 80 blood samples were obtained from donors at the Transfusion Medicine Unit, HUSM. The ethnic origins of the donors were Malays (n=73, 91.3%) and non-Malays (n=7, 8.75%). Males comprised 88.1% of the donors. Thalassemia was detected in 16.25% (n=13) of the blood donors. Of those with thalassemia, 46.2% (6/13) were anemic. Microcytosis and hypochromia were detected in 84.6% (n=l1) and 84.6% (n=l1) of these donors, respectively. The types of thalassemias detected were Hb E, 11.25% (n=9/80) and beta thalassemia trait, 5% (n=4/80). Among the thalassemias detected, the Hb E hemoglobinopathy was comprised of Hb E/ alpha-thalassemia (38.5%: n=5), Hb E /beta-thalassemia (23.1%: n=3), Hb E trait (7.6%: n=1) and beta-thalassemia (30.8%: n=4). In conclusion, screening for thalassemia trait should be included as part of a standard blood testing before blood donation. Further studies are required to look at the effects of donated thalassemic blood. PMID- 17120979 TI - The prevalence and determinants of iron deficiency anemia in rural Thai-Muslim pregnant women in Pattani Province. AB - This study was conducted in order to describe the type of anemia and risk factors for iron deficiency anemia in Pattani Province, Thailand. A cross-sectional survey was conducted from March to October 1997 in five randomly selected districts, choosing villages in the catchment area of a random sample of 30 out of 57 health centers (HC). All resident eligible pregnant women (PW) at 32-40 weeks of gestation without any overt diseases were selected. Food intake and antenatal health history were assessed by a food frequency questionnaire, health questionnaire and a review of HC records. Of the 180 enrolled PW, the prevalence of iron deficiency (ID), iron deficiency anemia (IDA) and other anemia were 34.4, 37.8 and 7.8%, respectively. PW in the last group were excluded from the analysis of predictors of iron status. Stool samples were obtained from 130 PW. The prevalences of hookworm, Ascaris and Trichuris were 47, 48 and 25 %, respectively. The number of ante-natal care (ANC) visits ranged from 0-8 with a median of 3 visits. Of those PW who visited, 97% reported receiving iron tablets. The compliance rate with iron tablets was low especially in the third trimester (9-12 %). Ordinal logistic regression showed that the risks for ID and IDA were reduced with statistical significance at a gestational age greater than 34 weeks, with more than three ANC visits, and increased consumption of meat and calories, but increased with hookworm infection. Compliance with iron tablet supplementation did not significantly reduce the risk for ID and IDA. In this study, PW had high percentages of ID and IDA. The risk factors identified in this report require intervention to eliminate them. PMID- 17120980 TI - A SPINK1 gene mutation in a Thai patient with fibrocalculous pancreatic diabetes. AB - Fibrocalculous pancreatitis diabetes (FCPD), a late stage of tropical chronic pancreatitis (TCP), is classified as a secondary cause of diabetes mellitus resulting from pancreatic exocrine dysfunction. The distinctive features of FCPD and TCP are young age at onset, presence of large intraductal pancreatic calculi, and reported mainly in tropical developing countries. Their etiology is still obscure, but the autodigestion due to aberrant intraductal activation of zymogens by trypsin is thought to be a primary common event. Recently, mutations in SPINKI gene encoding a pancreatic secretory trypsin inhibitor have been reported in association with an increased risk of pancreatitis. We describe a heterozygous mutation, IVS3+2 T>C, of SPINK1 gene in a young Thai female patient with typical presentation of FCPD. To our knowledge, this is the first report of the SPINK1 gene mutation in a FCPD patient in Southeast Asia. PMID- 17120981 TI - Necrotizing non-granulomatous lymphadenitis: a clinicopathologic study of 40 Thai patients. AB - The purpose of this study was to describe the clinicopathological features of 40 cases of necrotizing non-granulomatous lymphadenitis in Thai patients. The clinical features, histomorphology and special stains were evaluated in 40 Thai patients from the pathology records of King Chulalongkorn Memorial Hospital from January 2001 to December 2003 in those diagnosed as having necrotizing non granulomatous lymphadenitis. Of the 40 patients, 17 cases (42.5%) had Kikuchi Fujimoto disease (KFD), 8 cases (20%) had tuberculosis (TB) lymphadenitis and 1 case (2.5%) had systemic lupus erythematosus (SLE) with associated lymphadenitis. Fourteen cases (35%) did not have a specific diagnosis due to a lack of follow-up data. KFD most commonly occurs in young women, and is characterized by the presence of coagulative necrosis and karyorrhexis often centered in the paracortex, an absence of neutrophils and plasma cells, proliferation of various cells composed of lymphocytes, histiocytes, immunoblasts and plasmacytoid monocytes and the absence of a granuloma. Tuberculous lymphadenitis usually occurs in women with a mean age of 34.25 years. The lymph nodes reveal extensive coagulative necrosis involving the cortex, paracortex and medulla, proliferation of mixed inflammatory cells, including neutrophils, lymphocytes and plasma cells in the necrotic area and the presence of proliferating histiocytes at the periphery of the necrotic area. The lymph nodes of SLE-associated lymphadenitis reveal large numbers of plasma cells and hematoxylin bodies. We suggest that necrotizing non-granulomatous lymphadenitis is not specific for any disease, but rather a common histologic change found in diseases, such as TB, SLE, and KFD. Further investigation to obtain a definite diagnosis should be done for appropriate treatment. PMID- 17120982 TI - Normal spirometry, gas transfer and lung volume values in Papua, Indonesia. AB - Spirometry is an efficient and clinically useful tool in the diagnosis and management of chronic lung disease. It relies on an appreciation of normal lung function that can vary between populations. In order to improve the utility of spirometry, gas transfer and lung volume measures for clinical and research use in Papua, Indonesia, we determined lung function in Papuan and non-Papuan Indonesian adults who did not have evidence of lung disease. A cross-sectional survey of Papuan and non-Papuan Indonesians 18 years or older with no history of chronic cough or recent wheeze was made. Spirometry, gas transfer and total lung capacity (TLC) were determined and regression models developed for normal values. The spirometry values were similar but not directly comparable to similar studies in Papua New Guinea populations. Papuan highland residents demonstrated independently greater values of gas transfer and total lung capacity in comparison to lowland Papuans. Values for lung function in apparent respiratory health were shown to differ between Papuan and non-Papuan Indonesian populations and in comparison to reference values derived from non-indonesian populations. Differences in age-related decline in lung function would suggest that simple proportional correction based on values derived from non-Indonesian populations may be inappropriate and would support the development of similar reference values in other populations. Whether differences seen here are innate or occur as a consequence of in-utero and post-partum environmental exposure remains to be accurately elucidated. PMID- 17120983 TI - High sister chromatid exchange among a sample of traffic policemen in Bangkok, Thailand. AB - There are several volatile substances from the traffic, including benzene, toluene, carbon monoxide, lead and formaldehyde. Most of these substances are considered carcinogens. Police are at occupational risk for toxic fume exposure. This study compared sister chromatid exchange (SCE), a marker for genotoxicity, among a sample of Thai traffic policemen in Bangkok with healthy control subjects. Thirty police officers (all male) and 20 controls were included in this study. The average (mean+/-SD) SCE for policemen and controls were 4.40+/ 0.93/cell and 0.24+/-0.12/cell, respectively. A significantly higher SCE among the policemen was observed. Concern for and prevention of toxic substance exposure in traffic police officers should be made a national goal. PMID- 17120984 TI - Tobacco and alcohol use among the youth of the agricultural tea industry in Assam, India. AB - A cross-sectional study was conducted to assess tobacco use (both smoking and non smoked tobacco) and alcohol use among tea garden youths of Assam, India during the period 2002-2003. A total of 650 tea plantation youth age 15-24 years (255 males, 395 females) from eight randomly selected tea plantations, Dibrugarh District, Assam, were interviewed to collect information on alcohol and tobacco use using a pre-designed, pre-tested questionnaire. Nearly 59% of the respondents had no formal education. Fifty-eight percent of the youth used at least one substance and 27.4% were concurrent users of both alcohol and tobacco. The smoking rate was only 2.2% (4.7% in males, 0.5% in females). However, 52.5% of the study population used non-smoked tobacco (56.9% males, 49.6% females). The prevalence of alcohol consumption was 32.2% (43.9% males, 24.6% females). A higher rate of alcohol and tobacco use was found among the respondents who had no formal education or were school dropouts. A higher rate of alcohol and tobacco use were seen among respondents in whom both parents were illiterate. Working as a manual worker in the tea industry is significantly associated (p<0.01) with higher rates of alcohol and tobacco use. We recommend a vigorous campaign against tobacco and alcohol use among tea plantation youth to reduce the health risks associated with the use of these two substances. PMID- 17120985 TI - Modified Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE) as a screening test for dementia for Thai elderly. AB - A potential test for early detection of dementia in the elderly is the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE), which is based on information from the informant for the elderly about the changes of the elderly in everyday cognitive functioning associated with dementia. The present study aimed to modify and assess the reliability and validity of the modified IQCODE consisting of 32 items. The study consisted of two methods of assessing dementia: DSMIV diagnosis carried out by clinicians, and informants responding to the IQCODE. The subjects were 200 pairs of elderly subjects and their informants who visited the Geriatric Clinic, Ramathibodi Hospital. The optimal cutoff score on the modified IQCODE was 3.42, with 90% sensitivity and 95% specificity. The positive predictive values, negative predictive values, and accuracy were 0.94, 0.90, and 0.92, respectively. The IQCODE items had high internal consistency. The IQCODE associated with the elderly person's age, but not with their gender and educational level; nor were they associated with the demographic characteristics of the informant. Therefore, the IQCODE could be used as an alternative screening test for dementia in Thailand with acceptable sensitivity and specificity. This tool may be useful for dementia screening in the community and the geriatric clinic for early detection of disease. PMID- 17120986 TI - Long stick exercise to improve muscular strength and flexibility in sedentary individuals. AB - The objective of this study was to investigate the effects of long stick exercise training on the strength and flexibility of sedentary individuals. An observational prospective study was conducted at the Prince of Songkla University. Eighty-five subjects who did not engage in any regular physical activity for at least three months before the study. A long stick exercise program was taught by a master for 45 minutes, at least three times weekly, for three months. Handgrip strength, back-leg strength, and flexibility were measured at the beginning of the program as a baseline, and at one, two, and three months of training. The subjects had increased back-leg strength and flexibility. Median back-leg strength increased from the baseline by 0.07 kg/weight, 0.19 kg/weight, and 0.21 kg/weight, at one, two, and three months, respectively (p< 0.05). Median flexibility (sit-and-reach test) improved from the baseline by 4.34 cm, 4.71 cm, and 5.56 cm, at one, two, and three months, respectively (p = 0.001). There were no statistically significant changes in handgrip strength. PMID- 17120987 TI - [Changes in range of mosquito-borne diseases affected by global climatic fluctuations]. AB - Climate models suggest the strong possibility of range increase of the diseases transmitted by parasitic arthropods, mostly mosquitoes. In predicting processes of malaria and Dengue diseases dispersion the estimation of risk is based mostly on reproduction rate of vector species. These models allow to calculate the critical threshold of host density which is necessary to maintain parasites and pathogens transmission. Such studies based on integrated mathematical modelling indicate widespread increase of risk due to expansion of the areas suitable for mosquito-borne diseases transmission. This predicted increase is the most pronounced at the borders of the endemic areas and at higher altitudes within malaria and Dengue areas. The simulated change in mosquito-borne diseases risk must be interpreted on the basis of local environmental conditions as well as the effects of socio-economic developments and control disease programs. Apart from mathematical models the sequencing of proteins and DNA of vectors and their pathogens as well as satellite technology (GIS) are taken into consideration. It is supposed that potential impact of global climate change on malaria and Dengue risk can be reduced by constant warning system based on biological monitoring of mosquito vector species and their pathogens. Efficient care system connected with full diagnosis, treatment and prophylaxis of transmission diseases are also required. PMID- 17120988 TI - [Tick-transmitted diseases which may threaten health of dogs in Poland]. AB - Tick-borne dog infectious diseases constitute an emerging problem in veterinary medicine in recent years. The diseases recognized as characteristic for tropical and semi-tropical regions are diagnosed in dogs in countries with temperate climate including Poland. The major reason for such phenomenon is that Polish tourists increasingly travel with their dogs into subtropical and tropical areas of the world. Coming back to Poland, dogs often bring in non-native species of ticks as well as pathogens transmitted by them. Tick-borne diseases can result from infection with bacteria, viruses and parasitic protozoa. In this review we present most important tick-transmitted diseases which may occur in dogs in Poland. PMID- 17120989 TI - [Entomopathogenic nematodes, insects, bacteria and their relationship used in practice]. AB - In the paper are presented entomopathogenic nematodes (EPN) as a vector of bacteria killing insects. Many abiotic and biotic factors influence the survival and the virulence of mentioned organisms. These biological agents of control are used in practice over twenty years. Knowledge about the relationships between EPN insect-bacteria allow to increase efficacy of treatment used in order to limiting populations of pests. The paper presents a short review of literature data of titled subjects. PMID- 17120990 TI - Helminth parasites of laboratory mice and rats. AB - Rodents, as mice and rats are the most common laboratory animals used in research and testing. They are seldom investigated for autochthonous ecto- and endoparasites prior their utilization in the experiments. Helminth parasites can alter the interpretation of final results. Pinworms commonly infecting laboratory rodents include mainly the mice pinworms Syphacia obvelata and Aspiculuris tetraptera, and in rats Syphacia muris. The fact that many laboratory rodent colonies were found to be parasite contaminated suggests a need for eradication and improvment of the quality of laboratory rodents. This review reports the data on the presence of helminth parasites in laboratory rodents colonies, and suggests to pay special attention on controlling the sanitary conditions of animal houses. PMID- 17120991 TI - Content of glycogen and trehalose and activity of alpha-amylase and trehalase in Galleria mellonella larvae infected with entomopathogenic nematodes Steinernema affinis and S. feltiae. AB - INTRODUCTION: The influence of infection with two species of entomopathogenic nematodes of Steinernematidae family on metabolism of glycogen and trehalose of the host was studied. MATERIAL AND METHODS: Last instar larvae (L7) of Galleria mellonella were experimentally infected with Steinernema affinis and S. feltiae. At 6, 12, 18 and 24 h after infection concentrations of trehalose and glycogen as well as activity of trehalase and alpha-amylase were determined. RESULTS: The content of glycogen was lower in insects infected with S. feltiae than in the controls and animals infected with S. affinis. The content of trehalose was higher in insects from both infected groups than in the controls. Its concentration was slightly higher in larvae infected with S. affinis than in those infected with S. feltiae. The activity of alpha-amylase after infection with S. affinis was low. It was significantly higher in insects infected with S. feltiae. In animals of both infected groups, following a significant reduction at 6 h, the activity of trehalase remained at a similar level, higher than in the controls. In the paper the effects of infection with (i) different species of entomopathogenic nematodes and (ii) the importance of the developmental stage of the insect-host for changes in its metabolism of glycogen and trehalose were discussed. PMID- 17120992 TI - [The analysis of somatic antigens extracted from Aspiculuris tetraptera (Oxyuridae) and their role in eliciting immune response in laboratory mice]. AB - BACKGROUND: The aim of this study was isolation and examination of Aspiculuris tetraptera somatic proteins and somatic antigens role in eliciting of immune response in laboratory mice. MATERIAL AND METHODS: In our investigation 40 laboratory mice (BALB/c strain) were used. To extract the somatic proteins Tris HCl buffer with NaCl and Triton X-100 was used. The analysis of somatic antigens was undertaken by Western blotting. RESULTS: The study showed the presence of 14 protein bands ranging from approximately 82 to 28 kDa. Glicoproteins detection revealed 13 bands in range between approximately 70 to 30 kDa. There was no reaction observed with immunoglobulins IgA. Comparision of these results with earlier studies concerning S. obvelata somatic antigens show that there are proteins and glicoproteins with the same molecular weights for both species. It is also observed that S. obvelata somatic extract is more diversed and have higher antigenicity than A. tetraptera. Hence, we may suppose this fact could favour easier colonization of the host by A. tetraptera. PMID- 17120993 TI - Parasite fauna of the eel, Anguilla anguilla (Linnaeus, 1758), from the Polish part of the Vistula Lagoon. AB - INTRODUCTION: Eel is one of the most valuable commercial fish species of the Vistula Lagoon. For this reason its infection by non-indigenous species of parasites is the subject of the interest of sciencists. MATERIAL AND METHODS: A total of 90 eel specimens caught in the Polish part of the Vistula Lagoon were examined over the period of May-August 2005. The fish were subjected to standard procedures of parasitological examination and parasite identification. The fish were divided into two length classes: smaller and larger than 50 cm. To assess the effects of parasites on fish condition, the Fulton coefficient, expressed with the formula: WF = G x L(-3) x 100, was calculated. RESULTS: The fish were found to be hosts to 17 parasite taxa. The Ciliophora were represented by Trichodinella epizootica (Raabe, 1950); the Myxozoa by Myxidium giardi Cepede, 1906; the Digenea by Bunodera luciopercae (Muller, 1784), Deropristis inflata (Molin, 1859), Diplostomum spp., and Ichthyocotylurus platycephalus (Creplin, 1852); the Cestoda by Bothriocephalus claviceps (Goeze, 1782), Proteocephalus macrocephalus (Creplin, 1815), and Proteocephalus sp.; the Nematoda by Anguillicola crassus Kuwahara, Niimi et Itagaki, 1974, Camallanus lacustris (Zoega, 1776), C. truncatus (Rudolphi, 1814), and Hysterothylacium aduncum (Rudolphi, 1802); the Acanthocephala by Acanthocephalus lucii (Muller, 1776) and Corynosoma strumosum (Rudolphi, 1802); the Hirudinea by Piscicola geometra (Linnaeus, 1761); and the Copepoda by Ergasilus sieboldi Nordmann, 1832. All fishes were infected, and the mean intensity of all parasites was 4.8 inds. The nematode Anguillicola crassus was the most frequent parasite (67.8%, 4.2 inds), represented by all developmental stages, but mostly by adult specimens. The fish condition coefficient was somewhat higher in the eel larger than 50 cm (0.1746) than in those smaller than 50 cm (0.1734); in addition, only Anguillicola crassus free fish showed a higher condition coefficient (0.1861), compared to the infected specimens (0.1715). PMID- 17120994 TI - Parasite fauna of the European eel (Anguilla anguilla L, 1758) from the Russian part of the Vistula Lagoon (Baltic Sea). AB - INTRODUCTION: The european eel is one of the major fishing objects of the Russian fishery zone of the Vistula Lagoon (Baltic Sea). This is the reason that its parasite fauna was studied within 1998-2005. MATERIAL AND METHODS: Totally 142 individuals of 35-81 cm in length were examined. The parasites collected from various organs were fixed and processed according to commonly accepted methods. RESULTS: Twenty two parasite species representing the following higher taxa: Coccidia (2), Oligohymenophorea (2), Myxosporea (5), Monogenea (1), Cestoda (2), Trematoda (2), Nematoda (4), Acanthocephala (2), and Crustacea (2) were recovered. The following main features of the eel parasitofauna were determined: high specificity, predominance of the parasites with simple life cycle, the presence of three invader species (Anguillicola crassus, Pseudodactylogyrus anguillae, and Paratenuisentis ambiguus). The latter species was found in the Vistula Lagoon for the first time. PMID- 17120995 TI - [Helminth fauna of rodents (Arvicolidae and Muridae) from the Kampinos National Park]. AB - BACKGROUND: The aim of the study was to examine the parasite fauna of rodents living in natural environment, but in close contact with human seats (small village at the area of the natural big forest). MATERIAL AND METHODS: 169 rodents from family Arvicolidae and Muridae were examined. The material comprised: Microtus arvalis--5 specimens, Mus musculus--131, Apodemus silvaticus--9, Apodemus flavicollis--8, Apodemus agrarius--14 and Micromys minutus--2 specimens, collected between April 2003 and July 2004. RESULTS: The examination of some internal organs (alimentary tract, liver, lungs, kidneys, heart, body cavity) revealed the occurence of 8 helminth species: 3 species of Cestoda (Paranoplocephala macrocephala, Hymenolepis diminuta, Taenia taeniaeformis) and 5 species of Nematoda (Heterakis spumosa, Aspiculuris tetraptera, Syphacia obvelata, Heligmosomoides polygyrum, Mastophorus muris), all in the alimentary tract (including liver). The helminth fauna of every rodent species was determined, as well as seasonal dynamics of their frequency. It was stated that only the species which realize the complex life cycle undergo some seasonal fluctuations of their abundance. Additionally the distribution of particular helminth species along the alimentary tract of their hosts was described. PMID- 17120996 TI - Red fox (Vulpes vulpes) a new host species for acanthocefalan Moniliformis moniliformis (Bremser, 1811) in Poland. AB - Acanthocephalan Moniliformis moniliformis is a parasite of rodents, rarely also reported from carnivorous mammals. One female specimen of this parasite has been found in the small intestine of red fox Vulpes vulpes. It is the first report about this species invading the red fox in Poland. PMID- 17120997 TI - [The occurrence of fasciola hepatica in chosen regions of Poland based on molecular and serological methods]. AB - Fasciolosis, caused by the liver fluke (Fasciola hepatica) is an important issue for both human and animal health. The disease evokes economic losses which are a consequence of impaired animal productivity leading to higher costs of meat and milk production, as well as liver condemnation. The goals of this thesis were to: (1) elaborate a molecular method--PCR for the detection of F. hepatica DNA in intermediate and definite hosts; (2) estimate the usefulness of a recombinated cysteine proteinase produced in E. coli in the form of inclusive bodies in serological diagnosis of F. hepatica infection in definite hosts, using an enzyme linked immunosorbent assay (ELISA); (3) conduct field research on the prevalence of infection among intermediate and definitive hosts (cattle) in chosen regions of Poland, utilizing the elaborated methods. Based on the results obtained in this study, it was established that it is possible to detect F. hepatica DNA in the feces of definite hosts with the elaborated PCR method. The amplification of a 124 base pair tandem repeat allows the detection of fluke larval stages in intermediate hosts within 12 hours of exposure and F. hepatica infection in definite hosts (by the 5th week in rats, 8th week in sheep and 10th week in cattle). Therefore, the PCR test is more sensitive than traditional microscopic methods. Furthermore, it was determined that, the recombinated cysteine proteinase in the form of inclusive bodies, after solubillization exhibits antigenic properties of the native protein and the ELISA method based on this antigen may be useful as a tool for diagnosing fasciolosis in sheep and cattle, in both serum and milk samples. The test achieves a greater sensitivity and specificity than an ELISA based on native excretory-secretory antigens. The results of field research indicate that Fasciola hepatica is a frequent parasite of cattle in central and eastern Poland. The mean prevalence was 34.86% (+/- 16.95) in all studied areas. The prevalence among intermediate hosts varied greatly (0-100%). The elaborated tests were proved to be valuable, mutually complementing diagnostic tools, applicable to different epidemiological situations. PMID- 17120998 TI - [Waste management in the Republic of Croatia--current status]. AB - Waste management is one of the key ecological challenges of the modern world. Waste is a direct consequence of human activity and the quantity of generated waste is often an indicator of the economic strength and development of a community. This paper presents the figures indicating quantities of waste generated in the E.U. countries and Croatia. The framework of the waste management system is determined by law, governing bodies and institutions. In Croatia, the governing bodies include the parliament, relevant ministries, county and town administration, and local offices for environmental protection, all within their authorities and responsibilities. Current waste management in Croatia is characterised by the lack of accurate information about the quantity of waste produced, who produces what type of waste in what quantities, how it is further treated and disposed; then by inadequate treatment of waste, by the lack of adequate facilities within waste management system (treatment, disposal); by difficulties in finding appropriate location for disposal sites (difficulties in obtaining approvals by local communities and permits by relevant authorities). Only recently a database of dumps has been established. The regulatory framework is relatively good in Croatia, and in spite of problems, there is a growing activity and interest in waste management. PMID- 17120999 TI - [Waste management strategy and the role of the environmental protection and energy efficiency fund]. AB - On 14 October 2005, the Croatian Parliament adopted the National Strategy of Waste Management pursuant to Article 8 of the Waste Act. This strategy defines its objectives and the means to reach them, and gives an overview of the current situation in waste management. It seeks to provide a realistic framework for the effective reduction of national waste load and for a sustainable waste management. This Strategy defines 15 major waste flows which are in concordance with the EU practice. It also defines waste management buildings and plants, and looks into the remediation of municipal waste landfills. According to the Strategy, waste management will be present on all levels of administration: from national to local, and will include all areas of production, consumption, and everyday life. Founded on the EU's general principles of waste management, this Strategy provides the ideal framework for a sustained waste management in Croatia by the year 2025. Environmental Protection and Energy Efficiency Fund is one of the key administration bodies within the waste management system. It collects taxes for motor vehicles, for sulphur and nitrogen oxide pollution sources, for environmental hazardous and non-hazardous industrial waste load, and for the use of the environment. The Fund participates in the ongoing municipal landfill remediation projects and programmes all over Croatia (currently 161 landfills are being remedied) and helps to establish new regional and county waste management centres. It co-finances clean production technology projects, projects whose aim is to reduce the emission of harmful gases, projects for reusable energy sources and for more efficient use of energy, projects for remediation of unregulated municipal landfills, projects for primary recycling, and projects for biodiversity landscapes. According to an estimate, the implementation of the National Strategy of Waste Management will cost over 24 billion Croatian kunas (about 3.3 billion Euros). Beside national and local revenues, the Fund will play a key role in carrying this burden. PMID- 17121000 TI - [War consequences as the potential hazard for the Croatian karst ecosystem]. AB - In the recent war in Croatia, the karstic area of the country was endangered by hazardous waste, which calls for particular attention because of its exceptional ecological sensitivity. There are strong indications that various organic pollutants have entered the environment. It is also assumed that oil containing polychlorinated biphenyls (PCBs) was spilled in a number of locations. After the penetration in the environment, these substances are quickly absorbed in soil, sediment or bioaccumulated in the organisms of the aquatic ecosystem. In view of the locations of damaged electric transformer stations in Zadar, Bilice and Dubrovnik, there was some speculation on potential hazard posed by organohalogenated toxicants to Lake Vransko and the coastal area around Zadar, Sibenik and Dubrovnik. Samples of sediments and aquatic organisms were collected and analysed, and the results showed no significant or high levels of contaminants in the areas of Lake Vransko, Sibenik, and Dubrovnik (Petka location), while in the Mikulandra Bay near Sibenik, in Rijeka Dubrovacka, Brodanovo location and Marina near the Vruljica Creek in Zadar, significantly higher levels of PCBs were observed and these toxicants were additionally monitored within the APOPSBAL project from 2002 to 2005. The results of these investigations are presented in this paper. PMID- 17121001 TI - Behaviour of polychlorinated biphenyls contaminating soil near Zadar. AB - This study analysed PCB-contaminated natural soil excavated from an area adjacent to an electrical transformer station in Zadar, Croatia. For one year, PCB concentrations were measured in the soil and leachate water under natural climatic conditions. After 12 months, a negligible quantity of Aroclor 1248 (0.024% in average) and a total of seven key PCB congeners (IUPAC No: PCB-28, PCB 52, PCB-101, PCB-118, PCB-138, PCB-153, and PCB-180) were leached through the soils into water collected under lysimeters (0.032% in average). Although the amounts of leached PCBs were relatively small, their range 2 microg L(-1) to 15 microg L(-1) substantially exceeds the maximal allowed concentration of total PCBs in fishponds which is 1 ng L(-1). Soil samples were taken on two occasions from three depths from each lysimeter. The distribution of total PCBs as Aroclor 1248 equivalents and seven individual PCB congeners was determined within soil layers before and after planting seeds. In all soil plots the content of total seven PCBs and some individual PCB congeners in the surface soil layers significantly decreased. The total content of the seven PCB congeners in the surface soil layer of all soil plots decreased between 19.0% (soil plot 2) and 47.6% (soil plot 1) and in the middle soil layer between 8.1% (soil plot 4) and 37.4% (soil plot 1). PCB-28 and PCB-52 showed the highest percent of removal in all soil plots after 12 months of the experiment. The most important mechanism of removal of "lighter" congeners may be evaporation from the soil surface. Generally, our results showed that the PCBs accumulated in the deepest level of the soil, probably due to vertical transport and decreased evaporation. Some of the PCB content was lost to their degradation in the rhizosphere of the plants growing on the analysed soil plots. PMID- 17121002 TI - Organic contaminants in Croatian municipal wastewaters. AB - Municipal wastewaters are among the most important sources of waste materials released into the environment. In Croatia, this problem is especially pronounced since only 4.4% of the total wastewaters are subject to complete mechanical and biological treatment. Beside a limited number of regulated organic contaminants, wastewaters contain an extremely high number of different potentially harmful organic compounds. Comprehensive analyses of Croatian wastewaters using mass spectrometric techniques (GC/ MS, LC/MS) indicated the presence of a large number of different organic compounds with a predominance of two main groups of contaminants: petroleum hydrocarbons and detergent-derived surfactants. Recent investigations of specific organic contaminants in wastewater focused on the determination of so-called emerging contaminants, whose ecotoxicological relevance is based on new types of biological effects and for which water quality criteria have not yet been defined. The main goal of this paper is to make an overview of the present knowledge on the occurrence of different types of organic contaminants in Croatian municipal wastewaters, paying special attention to the emerging contaminants. PMID- 17121003 TI - [Organic contaminants in the Jakusevec landfill and their impact on groundwater quality]. AB - Landfilling is probably, the most popular disposal method for the management of domestic and industrial waste. Unfortunately, many landfills around the world do not include leachate collection systems and present a considerable risk to the underlying aquifers. There have been numerous reports on groundwater contamination in the vicinity of unprotected landfills, paying particular attention to specific organic contaminants. This problem is especially pronounced in landfills which, in addition to ordinary domestic refuse, contain waste of industrial origin. Jakusevec, the main landfill of the city of Zagreb, belongs to this category. The recently completed remediation programme for Jakusevec included comprehensive investigations with the aim to quantify various organic contaminants in solid waste and establish their possible infiltration into the underlying soils and groundwater. A detailed analyses using chromatographic and mass spectrometric techniques made it possible to identify a number of contaminants. According to their origin, the identified compound classes can be divided into the two main categories: markers of biological waste and its microbial transformation and markers of anthropogenic waste. Our investigations have shown that the composition of organic contaminants changes dramatically during the vertical transport from the solid waste through the unsaturated zone of soil to aquifer sediments, which involves both physicochemical and biotransformation processes. The vertical transport of polar organic contaminants was very efficient and led to a significant contamination of the aquifer in the vicinity of the landfill. PMID- 17121004 TI - [Air pollution at the waste disposal facility Jakusevec and surrounding populated area]. AB - The first air quality measurements in the communities of Jakusevec and Micevec surrounding the waste disposal facility Jakusevec started in the summer of 1995, that is, before the first section of the disposal facility underwent recovery. The continuous air monitoring was carried out at three measuring sites until the end of 2004. Over the years, air quality showed significant seasonal variations, with a tendency to have significantly higher concentrations of volatile compounds, especially hydrogen sulphite, mercaptans, mercury and gaseous fluorides in the summer than in the winter. In addition, pollution levels would rise substantially during recovery operations. The concentrations of H2S, mercaptans and gaseous fluorides would then exceed limit values defined by the Croatian law. Mercury levels kept below the limit value, but exceeded the recommended value. The concentrations of other pollutants were somewhat lower. PMID- 17121005 TI - Mercury in aquatic sediments and soils from Croatia. AB - Mercury is one of the most toxic and hazardous pollutant which occurs in the environment in different chemical forms, of which methylmercury is the most dangerous. Recently it was recognised that long-term anthropogenic inputs of mercury into environment resulted in the global mercury pollution and it was concluded that action should be taken to quantify the pollution sources and reduce human-generated releases of mercury. This paper presents new data on mercury levels in aquatic sediments from about 15 Croatian rivers, lakes and estuaries. It also brings data on mercury concentrations in soils from eight different regions of Croatia. Distribution of mercury species is discussed in more details for the Sava River, the Krka estuary and the Kastela Bay on the eastern Adriatic coast. Results show that sediments and soils from Croatia are generally not contaminated by mercury, except for some rivers and coastal locations under direct anthropogenic influence. PMID- 17121006 TI - [Radioactive waste due to electric power and mineral fertiliser production]. AB - Radiation Protection Unit of the Institute for Medical Research and Occupational Health in Zagreb has been conducting systematic investigations of radioactive contamination of the Croatian environment by anthropogenic fission products as well as by naturally occurring radioactive material (NORM) since 1963. Several critical sites in Croatia were identified for NORM, that is, for slag and ash repositories from coal-fired power plants and phosphogypsum repository from a mineral fertilizer production plant. As the coals and phosphate ores contain naturally occurring radionuclides, especially the members of the uranium and thorium radioactive chains, utilising these materials in various industries only enhances their natural radioactivity in residual waste. Consequently, the resulting activity concentrations of natural radionuclides in waste material could be several times higher than in the adjacent soil. These deposited materials pose permanent risk of radiation exposure due to the long physical half life of natural radionuclides (e.g., T 1/2 = 1600 years for 226Ra). Results of scientific investigations related to natural radioactivity are used in the recovery of slag and ash repositories and landfills, as well as in establishing regulatory criteria targeting import of coal and phosphate ores. In consequence, recently measured activity concentrations of natural radioactivity in imported materials used nowadays in coal-fired power plants are significantly lower than in previously used raw materials. Therefore, slag and ash can be used as additive materials in cement production. PMID- 17121007 TI - [Medical waste management: the law and its application]. AB - Medical waste management in Croatia is regulated by three fundamental legal acts: Waste Act, Ordinance on Waste Types and Directive on Manipulation of Waste Generated in Health Care. The implementation of these acts and waste management in general are also supported by Transport of Hazardous Materials Act and Toxic Substances Act. Modern trends in waste management promote an integral approach also called "waste chain". This integral approach to medical waste management is based on hierarchical structuring of waste management structure from the point at which the waste is produced to its final disposal. Priority is always given to reducing the amount and the harmfulness of waste by replacing harmful materials with those that are not, but where this is not possible, waste management includes reduction by sorting and separating, pretreatment on site, safe transportation, final treatment and sanitary disposal. Methods of choice for the final treatment should be the least harmful for human health and environment. Integral medical waste management could greatly reduce quantities of medical waste and consequently financial strains. Even though Croatian regulations define and cover all steps in the waste management chain their implementation is one of the country's greatest issues. Improper practice is seen from waste production in healthcare institutions to the final disposal, and includes handling, sorting, use of containers, or treatment in incinerators. Information on quantities, type and flow of medical waste are inadequate, just as sanitary control. In addition, Croatia lacks locations for sanitary landfills. Croatia has signed international conventions on the supervision and control of transfrontier waste shipments. Within the CARDS project, Croatia has started to harmonise its laws with acquis communitaire related to waste management. Although waste management is one of the priorities defined within the National Environment Action Plan, Croatia still has not made a strategy for waste management. PMID- 17121008 TI - Marine radioecology and waste management in the Adriatic. AB - This paper gives a review of marine radioecology research in the Adriatic area carried out by the Radiation Protection Unit of the Institute for Medical Research and Occupational Health. Measurements of radioactivity in the Adriatic started in 1963 as a part of an extended monitoring programme of radioactivity in Croatian environment. The main sources of radioactive contamination of the Adriatic Sea are the fallout from past nuclear weapon testing conducted in the atmosphere and the Chernobyl accident. In 2005, the activity concentrations of fission radionuclides were detectable at very low levels in all environmental samples collected on the Adriatic. The 90Sr data obtained from long-term monitoring were used to estimate the upper limit of the Adriatic seawater turnover time, which turned out to be (3.4 +/- 0.4) years. Detailed knowledge about seawater circulation, including the turnover time is essential for planning an overall communal and other wastewater management on the Adriatic coast. The paper concludes with the prospects for future marine radioecological investigations. PMID- 17121009 TI - [Environmental protection techniques used in the federal state of Baden Wurttemberg in Germany]. AB - Within the framework of the programme "Partnership for Sustainable Development" The Ministry of Environment of the German Federal State of Baden-Wurttemberg has come up with a project "Study Visit--Environmental Protection Techniques". It was intended as a three-week study visit for environmental protection experts from Central and Eastern European Countries (CEEC) to learn about the environmental protection techniques used in this federal state. Visits were paid to companies producing, applying or installing plants based on the state-of-the-art environmental protection techniques. The project started in 2005 and will last five years. The first visit to Baden-Wurttemberg was scheduled for 25 September 14 October 2005 for 12 experts from 12 countries: Bulgaria, Croatia, Cyprus, Czech Republic, Estonia, Hungary, Latvia, Lithuania, Poland, Romania, Slovakia and Ukraine. PMID- 17121010 TI - Report of the Poison Control Centre for the period 1 February-30 April, 2006. PMID- 17121011 TI - [Cloning, expression and purification of human nuclear autoantigenic sperm protein (hNASP) and preparation of its polyclonal antibody]. AB - OBJECTIVE: To acquire the purified human nuclear autoantigenic sperm protein (hNASP) and its polyclonal antibody for investigating the possible functions of hNASP involved in fertilization. METHODS: The coding sequence of hNASP gene was amplified from human testis RNA with specific primers, and the PCR product was cloned first into pMD-18T and then into pET-28a ( + ) after restriction digestion with BamH I and Hind III. The fusion protein was expressed in E. coli BL21 (DE3) after induction with IPTG. The recombinant protein NASP was purified from the supernatant with Ni2 -NTA His-bind resin under native conditions. RESULTS: The results of DNA sequencing and SDS-PAGE analysis showed the protein to be what we had hoped to acquire. ELISA showed that we had acquired rabbit anti-hNASP serum with high titer. CONCLUSION: High purity recombinant hNASP protein could be obtained with the above-mentioned prokaryotic expression method, and so could the rabbit anti-hNASP serum with high titer and high specificity. PMID- 17121012 TI - [Role of MAPK signal transduction pathway phosphorylation induced by granulocyte macrophage-colony stimulating factor in the proliferation of hormone-independent prostate cancer]. AB - OBJECTIVE: To investigate the phosphorylation intensity of MAPK pathway molecular Erk1/2 and the proliferation of prostate cancer cell line PC-3M. METHODS: Flow cytometry and RT-PCR were employed to study the ratio of different cell cycles and phases, respectively, before and after GM-CSF stimulation. Erk1/2 phosphorylation intensity was examined by Western blot simultaneously. RESULTS: The rate of PC-3M cells at S and G2/M stages and the expression intensity of Ki 67 increased after GM-CSF incubation in a dose-dependent manner. The phosphorylation intensity of Erk1/2 increased remarkably after stimulation with GM-CSF. CONCLUSION: The intensification of Erk1/2 phosphorylation is one important molecular mechanism of the proliferation of hormone-independent prostate cancer. PMID- 17121013 TI - [Expression and role of nitric oxide synthase in the testis and epididymis of Macaca fascicularis]. AB - OBJECTIVE: To investigate the expression and the role of nitric oxide synthase (NOS) in the testis and epididymis of macaca fascicularis. METHODS: The immunohistochemical ABC method was used to observe the localization of nitric oxide synthase in the testis and epididymis of the macaca fascicularis. RESULTS: (1) nNOS immunoreactivity was found in the spermatogenic cells of seminiferous tubules, the epithelia of epididymal efferent ducts, sperm and the endothelia of blood vessels; (2) iNOS was expressed in the epididymal efferent duct, the sperm inside the duct, and the myoid cells and endothelia of blood vessels; (3) eNOS immunoreactivity was detected in the interstitial cells of the testis, the epididymal efferent duct, the sperm inside the duct, and the myoid cells and endothelia of blood vessels. CONCLUSION: NOS is extensively expressed in the testis and epididymis of the macaca fascicularis and it may play an important role in such processes as spermatogenesis, sperm maturation and testosterone secretion. PMID- 17121014 TI - [Evaluation of the determination of seminal ACP and gamma-GT activities and correlation between seminal ACP or gamma-GT activity and semen parameters]. AB - OBJECTIVE: To evaluate the determination of seminal acid phosphatase (ACP) and gamma-glutamyltranspeptidase (gamma-GT) activity, and analyze the correlation between seminal ACP or gamma-GT and semen parameters. METHODS: ACP and gamma-GT activities in 133 samples of seminal plasma were measured. Two of the samples were randomly selected for intra-assay, one for the detection of ACP activity and the other for gamma-GT activity. And another four were selected the same way for the same purpose, two for the detection of ACP activity and the other two for gamma-GT activity. The semen volume, pH, sperm concentration, motility, and grade a and -b motility were analyzed by CASA system and so were the correlation between seminal ACP or gamma-GT activity and semen parameters. RESULTS: There was significant positive correlation between ACP and gamma-GT activities (r = 0.570, P = 0.000). The intra-CV of ACP was 13.72%, and inter-CVs of ACP were 13.80% and 15.49%. The intra-CV of gamma-GT was 7.68%, and inter-CVs of gamma-GT were 7.76% and 9.73%. Both seminal ACP and gamma-GT activities had significant negative correlation with pH (r = -0.330, P = 0.000 vs r = - 0. 388, P = 0.000). There was obvious correlation between gamma-GT activity and sperm concentration (r = 0.165, P = 0.045), but not between ACP activity and sperm concentration (r = 0.048, P = 0.546). Neither of seminal ACP and gamma-GT activity was correlated with sperm motility, grade-a and -b motility, semen volume, abstinence time and age. CONCLUSION: The precision of the measurement of gamma-GT activity in seminal plasma was higher than that of ACP. The correlation between seminal gamma-GT activity and semen parameters was similar to that between seminal ACP activity and semen parameters. Thus, the determination of gamma-GT activity was a more reliable marker than that of ACP activity for the evaluation of prostate function. PMID- 17121015 TI - [Meiosis related gene expression in rat spermatogenesis]. AB - OBJECTIVE: To find the expression of specific genes related to the meiosis of germ cells during spermatogenesis in the rat testis. METHODS: Segments of seminiferous tubules were obtained from the adult male SD rats, at stages XIII - I of meiosis, and the interstitial cells of the same testis were isolated under the stereomicroscope. The total RNAs of stages XIII - I segments and the testicular interstitial cells were extracted respectively, and mRNA differential display RT-PCR (DDRT-PCR) was conducted. The obtained cDNA fractions were purified and recovered, the reverse dot blot hybridization, sub-clones and screens of blue/white dots performed, and the results of sub-clones were identified by restriction endonuclease EcoR I digestion. RESULTS: Sixteen differential cDNA fractions were obtained through primary DDRT-PCR, 7 from stages XIII - I segments and 9 from the testicular interstitial cells. Another 11 were selected for further screening by reverse dot blot hybridization, their size ranging from 200 to 500 bp, of which 6 were from the stages XIII - I segments of seminiferous tubules and the other 5 from the rat testicular interstitial cells. All of the 11 cDNA fractions were sub-cloned and screened by blue/white dots. CONCLUSION: Specifically expressed differential cDNA fractions can be obtained and primarily identified from testicular interstitial cells and the seminiferous tubules, which, as the sequence tags of the testicular meiotic expression, deserve further investigation. PMID- 17121016 TI - [Diagnosis and treatment of testicular torsion in children]. AB - OBJECTIVE: To explore the early diagnosis of testicular torsion in children. METHODS: The clinical data of 24 patients with testicular torsion were analyzed retrospectively. RESULTS: The patients ranged in age from 4 months to 15 years ( mean 10.3 years), 17 left and 7 right testes involved. The duration between the onset and operation varied from 1 hour to 4 months. Diagnoses were initially and correctly made in 16 cases and delayed in 8. Surgical explorations were carried out in 23 cases, and resection of the testis performed in 17. CONCLUSION: Testicular necrosis is associated not only with the duration and degree, but also with the extent of the torsion. Color Doppler has clinical value in the early diagnosis of child testicular torsion. Timely surgical exploration should be performed for cases of acute scrotal problem suspected of child testicular torsion. PMID- 17121017 TI - [Inhibition of survivin gene expression in PC-3 cells by RNAi]. AB - OBJECTIVE: To construct eukaryotic expression vectors by using the pSilencer3. 1 H1 neo vector for inhibiting human survivin gene by RNA interference, and to detect the effect of the silenced survivin gene on PC-3 cells. METHODS: Three target gene segments were synthesized and cloned into the pSilencer3. 1-H1 neo vector respectively to construct three recombinant eukaryotic expression vectors: pSilencer3. 1-SVV1, pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3, which were identified by enzyme digestion analysis and DNA sequencing. Then the PC-3 cells were transfected with the recombinant vectors and the interference effect detected by RT-PCR, Western blot and immunohistochemical staining. The apoptosis index of the PC-3 cells was detected by flow cytometry and their proliferation detected by MTT method. RESULTS: Enzyme digestion analysis and DNA sequencing showed that three target segments were cloned into pSilencer3. 1-H1-neo vectors. The results of RT-PCR, Western blot and immunohistochemical staining indicated that pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3 vectors could knock down the transcription and expression of survivin gene. After transfected with pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3 vectors, the apoptosis index of the PC-3 cells was increased by 10% - 15% and their growth obviously slowly down. CONCLUSION: The transcription and expression of survivin gene were inhibited effectively by the recombinant eukaryotic expression vectors (pSilencer3. 1-SVV2 and pSilencer3. 1 SVV3) in the prostate cancer cell line PC-3. After transfected with pSilencer3. 1 SVV2 and pSilencer3. 1-SVV3 vectors, the apoptosis index of the PC-3 cells was increased and their growth inhibited. PMID- 17121018 TI - [Expression patterns of sonic hedgehog signaling molecules in human fetal prostate development]. AB - OBJECTIVE: To investigate whether the sonic hedgehog signaling pathway is involved in the development of human fetal prostate, and to evaluate the changing staining patterns of its molecules, sonic hedgehog (SHH), patchedl (PTC1), smoothened (SMO), and GLI1, in the human fetal prostate at various gestation stages. METHODS: Fifteen human fetal prostate specimens at various developmental stages (10 - 39 weeks) were included in this study. SHH, PTC1, SMO and GLI1 were detected in all the specimens by immunohistochemical technique. All the slides were observed and assessed under the light microscope. RESULTS: SHH, PTC1, SMO and GLI1 could be detected in human fetal prostate tissues, and their expression formed two surges, the former at week 16, and the latter at week 28. The staining of SHH and SMO was distributed only in the ductal epithelium but not in the stroma. The expression of PTC1 and GLI1 could be found mainly in the epithelium, with minimal staining in the stroma. CONCLUSION: The sonic hedgehog signaling pathway is involved in the development of the human fetal prostate. The high expression of its molecules at early gestation stages might be associated with the induction of prostatic buds, while their abundant expression at later gestation stages might be related to the prostate ductal branching, growth, differentiation and morphogenesis. PMID- 17121019 TI - [Characterization of the mRNA profile in ejaculated spermatozoa from healthy fertile men]. AB - OBJECTIVE: To explore the complexity of mRNA in the ejaculated sperm from healthy fertile men. METHODS: Semen samples were collected from 10 healthy fathers. The swim-up method was adopted to purify the sperm from possible contamination of somatic cells and the spermatozoal total RNA extracted by Trizol was used for SAGE library analysis. RESULTS: A totle of 21 052 SAGE raw tags were sequenced from 877 clones and 2 712 unique tags that occurred at least twice in the library were given further analysis. 19.7% of the unique tags had no match in the existing SAGE map, representing novel genes. Molecular function analysis revealed 67% of unique tags related to protein binding or nucleic acid binding categories, 41% to catalytic activity, 13% to message transducer activity, and 10% to transporter, structural and transcription regulator activity, respectively. CONCLUSION: There exists a complex repertoire of mRNAs in the ejaculated spermatozoa from fertile men. PMID- 17121020 TI - [Combined assay of serum prostate specific antigen and chromogranin A helps diagnosis of prostate cancer]. AB - OBJECTIVE: To evaluate the significance of the combined assay of chromogranin A (CgA) and prostate specific antigen (PSA) in the diagnosis of prostate cancer. METHODS: Serum CgA and PSA were detected by ELISA technique in 55 cases of prostate cancer (PCa), 25 cases of benign prostate hyperplasia (BPH), and 50 cases of normal subjects (control). RESULTS: The serum CgA level in the PCa group was significantly higher than those in the control and BPH groups (P < 0.05), and increased with clinical stages. The parallel and serial tests associated with serum PSA and CgA raised the rate of detection of prostate cancer. CONCLUSION: The combined assay of serum PSA and CgA is of significant clinical value in raising the rate of diagnosis of prostate cancer, as well as in staging and prognosing the disease. PMID- 17121021 TI - [Diagnosis and treatment of iatrogenic ejaculatory duct obstruction following prostatic hyperthermia (report of 3 cases)]. AB - OBJECTIVE: To investigate the characteristics of diagnosis and treatment of iatrogenic ejaculatory duct obstruction following prostatic hyperthermia. METHODS: Retrospective analyses were made of the clinical data of 3 cases of ejaculatory duct obstruction following prostatic hyperthermia. RESULTS: The 3 cases were diagnosed as urethrostenosis and deformity of the posterior urethra by transurethral ultrasound and semen analysis. And all the 3 patients had undergone prostatic hyperthermia for prostatitis. Transurethral resection of the ejaculatory duct (TURED) was performed to remove the obstruction and the postoperative semen analysis showed both semen volume and sperm count to be normal. CONCLUSION: Urethra microwave thermotherapy, urethra radiofrequency, or per urethra rheophore ablation can be adopted in the treatment of protatitis, but should be strictly indicated and cautiously selected lest secondary iatrogenic ejaculatory duct obstruction should result. For the treatment of this obstruction, TURED is the first choice. PMID- 17121022 TI - [Molecular identification of Ca2+ channels in human ejaculated sperm]. AB - OBJECTIVE: To identify the types of voltage-dependent Ca2+ channels (VDCCs) in human ejaculated sperm using molecular technology. METHODS: Based on the WHO criteria, the donated semen was screened by computer-aided sperm analysis (CASA), the sperm optimized by swim-up method, and the VDCCs' alpha1 subunits examined by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Such mRNA messages as alpha1H, alpha1G, alpha1E, alpha1B and alpha1C were detected, but alpha1A and alpha1D were not. CONCLUSION: Human motile sperms express multiple VDCC RNAs, among which T-type and non-L-type channel messages are likely to be predominantly expressed. Not only T-type but also non-L-type calcium channels may be major gates for the external calcium influx, which helps regulate sperm motility and acrosome reaction. PMID- 17121023 TI - [Type IV collagenase expression in human prostate carcinoma cell lines with different metastasis potentials]. AB - OBJECTIVE: To investigate the correlation between type IV collagenase expression and metastatic potential of human prostate carcinoma cells. METHODS: Human prostate carcinoma cell lines (LNCaP and C4-2B) with different metastatic potentials were selected. Using SABC immunocytochemical staining and zymographic analysis, we detected the difference of type IV collagenase expression and activity between the two cell lines. RESULTS: The expression of type IV collagenase (MMP-2, MMP-9) could be detected in both LNCaP and C4-2B, mainly in cytoplasm, much higher in C4-2B than in LNCaP (P < 0.01). The activities of type IV collagenase in the conditioned medium of prostatic carcinoma cell LNCaP was the lowest, and the quantitative estimation values of MMP-2 and MMP-9 were 0.89 +/- 0.02 and 0.86 +/- 0.01, respectively. However, the quantitative estimation values of MMP-2 and MMP-9 in the conditioned medium of C4-2B were 96.32 +/- 4.36 and 33.89 +/- 1.84, respectively. Compared with LNCaP, the activity of type IV collagenase in C4-2B was much higher (P < 0.01). CONCLUSION: Compared with LNCaP, the higher production capability of type IV collagenase in C4-2B may possibly contribute to its invasive and metastatic potentials. The expression of type IV collagenase of human prostate carcinoma cells is closely correlated to the metastatic and invasive potential. PMID- 17121024 TI - [Role of bFGF and TGF-beta1 in primary cultured prostatic stromal cells]. AB - OBJECTIVE: To study the role of the basic fibroblast growth factor (bFGF) and transforming growth factor beta1 (TGF-beta1) in benign prostatic hyperplasia (BPH). METHODS: The human stromal cells of BPH were isolated and cultured. The proliferation of the stromal cells cultured in serum-free medium was detected by MTT method, the phenotype changes of smooth muscle cells detected by immunohistochemical method, and the effect of different concentrations of bFGF and TGF-beta1 on the cultured stromal cells of BPH observed. RESULTS: bFGF stimulated the cultured BPH stromal cell proliferation (P < 0.05, P < 0.01) and decreased the expression of smooth muscle cell (SMC) phenotype in higher concentration (10 microg/L). TGF-beta1 (> 1 microg/L) inhibited stromal cell proliferation and increased the expression of SMC phenotype (P < 0.05, P < 0.01). 5 microg/ml bFGF and TGF-beta1 (0.001 microg/L, 0.01 microg/L) promoted stromal cell proliferation (P < 0.01), while 5 microg/L bFGF and TGF-beta1 (0.1 microg/L, 1 microg/L, 10 microg/L) inhibited it, slightly in 0.1 microg/L (P > 0.05) and significantly in 1 microg/L and 10 microg/L (P < 0.01), and increased the expression of SMC phenotype in higher concentration (> 1 microg/L, P < 0.01). CONCLUSION: bFGF stimulates the proliferation of the prostatic stromal cells of BPH in a time- and dose-dependent fashion and decreases the expression of SMC phenotype, TGF-beta1 inhibits the growth of stromal cells and induces the differentiation of stromal cells to SMC, both playing an important role in the mechanism of BPH. PMID- 17121025 TI - [Clinical features and epidemiological survey of perianal warts in 72 males]. AB - OBJECTIVE: To investigate the clinical features and epidemiological data of 72 cases of male perianal warts. METHODS: Seventy-two cases of perianal warts in our clinic dated from June, 2004 to April, 2006 were enrolled in the study, whose clinical information and epidemiological data were collected and analyzed. RESULTS: Perianal warts were most commonly seen in young and middle-aged men aged from 18 to 45, only 12.5% of whom had homosexual behaviors. Sauna was another predisposing factor of perianal warts in males in China (chi2 = 5.03, P < 0.05). Primary eruptions of the anus and rectum, like perianal pruritus, eczema, anus fissure, and haemorrhoids, often impaired the local integrity of skin/mucosa. Classical condyloma acuminate was found in 61 (84.72%) of patients, who were susceptive to the infections of HPV 6/11, and were flat condylomas related to HPV16/18. Cryotherapy was believed to be one of the most efficient therapeutic choices for flat perianal warts. Suppression of cellular immune response was identified in the patients by comparison between the subgroups of peripheral T cells and the normal control. CONCLUSION: Sauna is an essential predisposing factor of perianal warts in males, while anus sexual intercourse is not the main route of HPV infection. Classical condylomata acuminate constitute the majority of the eruptions, and flat condylomata come next. The study also provides some useful data for understanding the clinical and epidemiological features of perianal warts in Chinese males for the sake of prevention and treatment of the disease. PMID- 17121026 TI - [Color Doppler in differential diagnosis of scrotal mass]. AB - OBJECTIVE: To evaluate the differential diagnosis of scrotal mass with color Doppler ultrasound. METHODS: Retrospective analysis was made of 21 cases of scrotal mass confirmed both surgically and pathologically in our hospital. RESULTS: Eight of the total number were malignancy of the testis origin, accounting for 38.1% of whole study group and 13 were benign, accounting for 61.9%. Of the 13 benign cases, only 2 were of the testis origin (15.4%) while the other 11 (84.6%) were not. CONCLUSION: Color Doppler ultrasound plays an increasingly important role in the differential diagnosis of scrotal mass. PMID- 17121027 TI - [Extraperitoneal laparoscopic radical prostatectomy (report of 5 cases)]. AB - OBJECTIVE: To discuss the clinical experience of laparoscopic radical prostatectomy by extraperitoneal approach. METHODS: Five patients with localized prostate cancer underwent laparoscopic radical prostatectomy by extraperitoneal approach. The surgical procedure included the excision of the prostate, seminal vesicles, ampulla ductus deferentis and part of the bladder neck, followed by urethrovesical anastomosis. RESULTS: All the operations were successful. The mean operation time was 350 minutes (ranging from 270 to 420 mm); the mean estimated blood loss was 480 ml (ranging from 250 to 600 ml). The bowel activity was recovered with 48 hours after surgery. The patients were ambulant between the 2nd and 3rd postoperative days. The mean hospital stay was 8. 5 days (ranging from 7 to 12 days). The 3-8 months follow-up found no incontinence of urine; of the 3 preoperatively potent patients, 2 were able to have sexual intercourse; strictured stoma was reported in only 1 case. CONCLUSION: The extraperitoneal laparoscopic radical prostatectomy, keeping the procedure out of the peritoneal cavity, with small incision and rapid recovery, may be considered as a promising surgical method for patients with localized prostate cancer. PMID- 17121028 TI - [Ca2+ and sperm function]. AB - Ca2+ is an important positive ion in the living body. Recently, there have been quite a few reports about the function of Ca2+ in sperm. Calcium is considered as a regulator of sperm motility, a participant in sperm capacitation, and an essential second messenger for acrosome reaction. This paper reviews the relationship of Ca2+ with sperm function. PMID- 17121029 TI - [Present state of studies on anatomical anomalies in cryptorchidism]. AB - Cryptorchidism is a clinically common congenital deformity of the genital organ, the etiological factors of which are still not clarified up to now. Undescended testis is frequently accompanied with the anomalies of the spermatic cord, gubernaculum testis, processus vaginalis, and epididymis. Because these anatomical anomalies act as terminal factors or the ending in the links of the cause of cryptorchidism, it is of important directive significance for the clinical treatment of the problem to seek evidence of anatomical anomalies at orchiopexy to demonstrate the etiological factors of cryptorchidism. PMID- 17121030 TI - [Cavernous nerve reconstruction to restore erectile function following radical prostatectomy]. AB - Recent years have seen an increasing incidence of prostate cancer (PCa) and a more frequent occurrence of the disease in younger men. Damage to cavernous nerves caused by radical prostatectomy is the main reason for post-operative erectile dysfunction. So reconstruction of cavernous nerves at the time of radical prostatectomy may restore the patient's erectile function. At present, clinical studies of using autologous sural nerve grafts to bridge transected cavernous nerves have achieved modest success, and laparoscopic sural nerve grafting during radical prostatectomy has also been proved safe and feasible. Besides, various grafts have been used to reconstruct cavernous nerves in animal models, among which biodegradable conduits containing growth-promoting factors or cells are a promising option for the repair of damaged cavernous nerves. PMID- 17121031 TI - [Experimental research on the liver-qi stasis induced erectile dysfunction by mouse model]. AB - OBJECTIVE: To establish a mouse model of erectile dysfunction (ED) induced by liver-qi stasis for observing the changes in the mouse's sex organs and investigating the possible mechanism of ED. METHODS: Seventy-two Kunming mice, 24 males and 48 females, were randomly divided into a liver-qi stasis group, a severe liver-qi stasis group and a control group. A stressing box was used to exert gradual stress on the mice of the two experimental groups by confining them to the box and gradually reducing their room of movement so as to induce liver-qi stasis. In addition, the males of the severe liver-qi stasis group were further prohibited from any sexual activities to repress their sexual desire and intensify their liver-qi stasis. RESULTS: Obvious differences were observed in the autonomous behavior, sexual activity and body weight of the mice of the three groups. Pathological examination showed that the sex organ indexes descended in the liver-qi stasis group and even further in the severe liver-qi stasis group as compared with the controls. CONCLUSION: The stressing box can induce and intensify ED associated with liver-qi stasis. PMID- 17121032 TI - [Efficacy and safety of vardenafil in the treatment of erectile dysfunction in men with hypertension]. AB - The prevalence of erectile dysfunction (ED) is higher in hypertension patients than in non-hypertension men. Because doctors worry about the severe adverse events of drug combination, they tend to be reluctant to prescribe ED medicines for patients. Vardenafil, as a novel and highly selective phosphodiesterase 5 inhibitor, has been proved by many clinical trials to be quite safe for the cardiovascular system. A recent large-scale clinical trial showed that vardenafil could improve erectile function in ED men with hypertension, with sure safety and no significant clinical changes in the index of hypertension or heart rate. PMID- 17121033 TI - [Biomechanopharmacology annotation of action of promoting blood and removing blood-stasis]. PMID- 17121034 TI - [Correlations between diagnostic information and therapeutic efficacy in rheumatoid arthritis analyzed with decision tree model]. AB - OBJECTIVE: To explore the correlations between diagnostic information and therapeutic efficacy in rheumatoid arthritis (RA) with decision tree model analysis. METHODS: Three hundred and ninety seven patients came from 9 clinical centers were randomly divided into the Western medicine (WM) group (n=194) treated with non-steroidal anti-inflammatory drugs and slow-acting antirheumatic drug and the Chinese medicine (CM) group (n=203) with basic therapy and syndrome differentiation dependant TCM treatment. TCM and WM diagnostic information were collected. The ACR 20 was used for efficacy evaluation and the information of patients before treatment was analyzed by SAS 8.2 statistical package. Through single-factor exploratory analysis, odds ratio of efficacy and variable was calculated taken P < 0.2 as the including criteria for data mining analysis with decision tree model. All data were classified into the training set (75%) and verifying set (25%) with efficacy as the variable for layering to make further verification of the data-mining analysis. RESULTS: Twenty variables were included in the CM group and 26 in the WM group in the data-mining model. In the former, 9 variables were positively correlated to the efficacy, including degree of arthralgia, tenderness and morning stiffness, number of swollen joint, and joint with tenderness, levels of IgM, rheumatoid factor (RF), C-reactive protein (CRP), and total assessment from doctor; and disease duration and degree of nocturnal polyuria were negatively correlated to that. While in the latter, 8 were positively correlated to the efficacy, including erythrocyte sedimentation rate (ESR), sour and weak waist and knees, white fur in tongue, joint ache and stiffness, swollen joint, and total assessment from doctor and patient, and red tongue with yellow fur and leucocyte count negatively correlated to it. Data mining with decision tree analysis revealed that different combinations of morning stiffness, slight red tongue, joint tenderness and nocturnal polyuria in the CM group, and those of white fur in tongue, CRP level, leucocyte count and morning stiffness in the WM group showed different efficacy, which were also verified in the randomly chosen verifying set. CONCLUSION: To analyze the correlations between diagnostic information and therapeutic efficacy with decision tree analysis is conformed to the theory of TCM in applying treatment according to syndrome differentiation individually, thus it would contribute to elevate the accuracy of therapy. PMID- 17121035 TI - [Clincal observation of qingre yulin decoction in treatment of male infertility caused by accessory gland infection]. AB - OBJECTIVE: To study the effect of Qingre Yulin Decoction (QYD) on male infertility caused by accessory gland infection (AGI) with randomized controlled trial (RCT). METHODS: Sixty infertility outpatients were equally divided into two groups randomly, the QYD group treated with modified QYD and the control group with antibiotic plus vitamin E, both for 3 months with another 6 months' follow up. Pregnant rates, routine test of sperm and expressed prostatic secretion (EPS) were determined. RESULTS: The healed rate was 26.7% (8 cases), the markedly effective rate was 43.3% (13 cases), the effective rate was 16.7% (5 cases), and the total effective rate was 86.7% in the QYD group, while in the control group it was 6.7% (2), 30.0% (9), 40.0% (12) and 76.7% respectively, showing higher healed rate and total effective rate in the former than those in the latter. Sperm quality of infertility patients with AGI decreased obviously, manifesting short ened average liquefaction time, reduced concentration, survival rate and vitality of sperm. These abnormal changes were improved after treatment in both groups, and the efficacy was better in the QYD group than that in the control group. CONCLUSION: Infertility patients with AGI were manifested as oligospermatism and asthenospermia, which may not be the definite outcome of AGI. QYD is able to improve sperm quality, especially sperm vitality in infertility patients with AGI and therefore increase pregnant rate of their wives. PMID- 17121036 TI - [Effects of jingui shenqi pill combined prednisone on expression of glucocorticoid receptor and its clinical effect in treating bullous pemphigoid patients]. AB - OBJECTIVE: To investigate the effect of Jingui Shenqi Pill (JSP) on the expression of glucocorticoid receptor (GR) in the skin lesion and its clinical effect in treating bullous pemphigoid (BP) patients. METHODS: Thirty BP patients were randomly divided into the treatment group (n=15) treated with JSP plus prednisone and the prednisone group (n=15) with prednisone alone both for 4 weeks. And a normal control group was set up also. Expressions of GR-alpha and GR beta in the skin lesion of BP patients and the normal skin of the normal control were detected by immunohistochemical assay. Results The total effective rate was 93.33% in the treatment group, significantly higher than that in the prednisone group which was 73.33% (P < 0.05); GR-alpha expression was higher in the treatment group than that in other two groups (P < 0.01), while GR-beta expression in the treatment group was lower than that in the prednisone group (P < 0.01). CONCLUSION: JSP could increase GR-alpha expression and decrease GR-beta expression in the skin lesion of BP patients, so as to improve sensitivity of skin to glucocorticoid. PMID- 17121037 TI - [Relationship between pathological changes of coronary artery and severity of Xiongbi syndrome]. AB - OBJECTIVE: To study the relationship between the outcome of coronary angiography (CAG) and the severity of Xiongbi syndrome diagnosed according to "Guiding principle for clinical study on new traditional Chinese medicine in treating Xiongbi (angina pectoris)" (abbreviated as the principle). METHODS: Severity of Xiongbi syndrome in 207 patients were graded according to the principle and they were examined with CAG by Judkins methods. The relationship between the outcome of CAG and severity of Xiongbi syndrome was analyzed. RESULTS: Abnormal rate of ECG was markedly higher in patients with lesion involving 1 or 2 branches of coronary artery than that in patients without coronary artery lesion (P < 0.05), while no significant difference was found in that between patients with multiple branches lesions and those without lesion. Comparison of Xiongbi syndrome severity showed that there was remarkable difference between patients with normal coronary artery and those with lesions involving different branches (P < 0.01), but no difference among patients with 1 or 2 branches and those with multiple branches lesions; on the contrary, mild Xiongbi syndrome took up the highest rate (37%) in patients with multiple-branches lesions, and the rate was lower along with higher severity. CONCLUSION: There was no positive relationship between severity of Xiongbi syndrome and that of coronary artery disease. PMID- 17121038 TI - [Effects of xinshuai mistura on plasma angiotensin II, atrial natriuretic peptide and N terminal pro-brain natriuretic peptide in patients with heart failure]. AB - OBJECTIVE: To observe the effects of Xinshuai Mistura (XM) on plasma angiotensin II (Ang II), atrial natriuretic peptide (ANP) and N terminal pro- brain natriuretic peptide (Nt-proBNP) in patients with heart failure (HF). METHODS: Fifty-nine patients with HF (NYHA class II-IV) were randomly divided into two groups, the treated group treated with XM and the control group with Wuling Pulvis (WP), both on the base of captopril treatment. The treatment course was two weeks. Plasma levels of Ang II , ANP and Nt-proBNP were observed. RESULTS: The plasma levels of Ang I, ANP and Nt-proBNP showed an ascending tendency along with the increasing in severity of HF(P < 0.01), and significantly decreased after treatment in the treated group (P < 0.01), but with no remarkable difference as compared with those in the control group (P > 0.05). CONCLUSION: XM had definite therapeutic effects on excessive neuroendocrine activation in HF patients. PMID- 17121039 TI - [Clinical effect of shenbing mistura combined with glucocorticoid on recurrent nephrotic syndrome in children and levels of interleukin-6 and tumor necrosis factor-alpha in blood and urine]. AB - OBJECTIVE: To study the effect of Shenbing Mistura (SM) combined with glucocorticoid on recurrent nephrotic syndrome (RNS) in children and levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) in blood and urine. METHODS: The treatment group was treated with SM plus glucocorticoid, the control group with glucocorticoid alone, and a healthy control group was adopted, 30 cases in each group. The clinical effect and recurrence rate were observed, and levels of IL-6 and TNF-alpha in blood and urine were determined before treatment and at the 4th, 8th, 12th week after treatment. RESULTS: Significant difference of IL-6 and TNF-alpha levels in blood and urine was found in either the pre- and post- treatment auto-control of both the treatment group and control group, or in the inter-group comparison of them (P < 0.01); clinical effect also showed remarkable difference between the two groups (P < 0.01). Furthermore, the recurrence rate of the treatment group was lower than that of the control group showed by a 18-month follow-up (P < 0.01). CONCLUSION: SM combined with glucocorticoid could significantly reduce the recurrence rate and elevate the clinical effect in children with RNS, it could also lower the levels of IL-6 and TNF-alpha in patients' blood and urine. PMID- 17121040 TI - [Clinical study on colonic transmission time and the effect of sini powder on it in functional constipation patients]. AB - OBJECTIVE: To investigate the characteristic of colonic transmission in functional constipation (FC) and the effect of traditional Chinese medicine (TCM) Sini Powder (SP) on it. METHODS: The colonic transmission time (CTT) of 36 patients with FC (the FC group) and 22 healthy subjects (control group) was measured through colonic transmission test, and CTT of entire colon and that of various subsections was calculated with Hinton method and Arhan method respectively. After then, the FC group was treated with SP for 7 days, and CTT was detected again after treatment. RESULTS: Before treatment, body mass index (BMI) was higher, CTT of entire colon, left half colonic section, and sigmoid rectal section were longer in the FC group than those in the control group (P < 0.05), no statistical difference in CTT of right half colon was found between the two groups (P > 0.05). After FC patients being treated with SP, their CTT of whole colon, left half colonic section and sigmoid-rectal section were significantly shortened (P < 0.05). CONCLUSION: FC patients were characterized by increased BMI and CTT prolonged and unevenly distributed in subsections, especially in the left half colon, sigmoid and rectum; SP could shorten the CTT in FC patients. PMID- 17121041 TI - [Clinical research on effects of yunpi zhixie granule in children with diarrhea]. AB - OBJECTIVE: To investigate the clinical effect of Yunpi Zhixie Granule (YZG) on children's diarrhea. METHODS: Three hundred diarrhea cases were randomly divided into two groups equally, the treated group treated by YZG orally and the control group by smacta, both for 7 days. The clinical efficacy was observed, routine scatologic analysis, scatologic cultivation, rotavirus antigen and urinary D xylose content were detected. RESULTS: Clinical results showed the effective rate in treating the chronic or Pi-deficiency diarrhea as well as the negative conversion rate and improvement rate of routine scatologic analysis were all higher in the treated group than those in the control group (P < 0.05); while no significant difference was found in negative conversion rate of scatologic cultivation and rotavirus antigen and urinary D-xylose content after treatment, compared with those before treatment (P > 0.05). CONCLUSION: YZG was effective in treating children' s diarrhea, especially on chronic diarrhea and diarrhea of Pi deficiency type. PMID- 17121042 TI - [Effects of yishen huoxue decoction on proliferating cell nuclear antigen expression and extracellular matrix in 5/6 nephrectomized rats]. AB - OBJECTIVE: To investigate the effects and mechanisms of Yishen Huoxue decoction (YHD) on chronic renal failure (CRF) rats induced by 5/6 nephrectomy. METHODS: The glomerulosclerosis model was established by 5/6 nephrectomy in rats. Experimental animals were allocated into the normal group, the model group, the YHD group and the benazepril group. Urine protein of 24 h (UP) at the 6th and 12th weekend after operation, blood urea nitrogen (BUN) and creatinine (SCr), albumin (Alb) and haemoglobin (HB) at the 12th weekend were measured, renal pathology changes were examined with light microscope, the expressions of proliferating cell nuclear antigen (PCNA) and fibronectin (Fn) were examined by immunohistochemistry and mRNA expressions of connective tissue growth factor (CTGF) and plasminogen activator inhibitor-1 (PAI-1) by RT-PCR at the 12th weekend. RESULTS: Compared with those in the normal group, the levels of UP, BUN and SCr, the area of glomerular mesangial matrix, the FN deposition, PCNA expression in glomeruli and tubular interstitium and mRNA expressions of CTGF and PAI-1 were all significantly higher in the model group (P < 0.05). All the above mentioned indexes were lower in the YHD group than those in the model group (P < 0.05). PCNA positively expressed cells in glomeruli of the normal, model group, YHD group and benazepril group was 7.00 +/- 2.24,34.78 +/- 6.96,15.75 +/- 2.61 and 15.50 +/- 2.57 respectively, positively correlated to the expression of CTGF, PAI-1, FN and SCr level. CONCLUSION: YHD could delay the progression of CRF in 5/6 nephrectomized rats, and the mechanisms were mainly related to the inhibition on renal cell proliferation and it induced over-expression of cytokines, and accumulation of extracellular matrix. PMID- 17121043 TI - [Effects of huanglian jiedu decoction on signal transduction of insulin receptor and insulin receptor substrate in adipose tissue of insulin resistant rats]. AB - OBJECTIVE: To investigate the effects of Huanglian Jiedu decoction (HLJD) on protein expressions and tyrosine phosphorylation levels of insulin receptor (InsR) and insulin receptor substrate-1 (IRS-1) in adipose tissue of insulin resistant (IR) rats, and explore its possible molecular mechanism in improving IR. METHODS: IR model was induced by intravenous injection of a small dose of streptozotocin combined with high fat and caloricity diet feeding in Wistar rats. And the model rats in the testing group were treated with HJD for 10 weeks. Fasting serum glucose and insulin were determined, and protein expressions and tyrosine phosphorylation levels of InsR and IRS-1 in adipose tissue of epididymides were detected by immunoprecipitation and Western blot. RESULTS: The protein expression of IRS-1 and the tyrosine phosphorylation levels of InsR and IRS-1 increased significantly in model rats treated with HLJD, compared with those in the untreated model rats. CONCLUSION: HLJD could increase the tyrosine phosphorylation levels of InsR and IRS-1 in adipose tissue in IR rats, which maybe one of its mechanisms in lowering blood glucose and improving insulin sensitivity of the target tissues. PMID- 17121044 TI - [Effect of the disassembled recipes of dahuang zhechong pill on proliferation and apoptosis of vascular smooth muscle cells in atherosclerotic rabbits]. AB - OBJECTIVE: To study the effects of the disassembled recipes of Dahuang Zhechong Pill (DRDZP) on proliferation and apoptosis of vascular smooth muscle cell (VSMC) of thoracic aorta in atherosclerotic rabbits. METHODS: The atherosclerotic rabbit model was induced by high-cholesterol diet and immune injury of endothelium. Expression of proliferating cell nuclear antigen (PCNA) in VSMC was detected by SP immunohistochemical technique, and VSMC apoptosis was observed with TUNEL technique. RESULTS: Compared with the model control, PCNA expression in VSMC decreased, while apoptotic cells increased significantly in all the groups treated by DR-DZP (P < 0.01). CONCLUSION: All the DR-DZP could inhibit VSMC proliferation and promote its apoptosis to modulate the balance between cell proliferation and apoptosis, so as to exert antiatherosclerotic action, among which the disassembled-recipe I is the main composition to contribute to antiatherosclerotic action of Dahuang Zhechong Pill. PMID- 17121045 TI - [Effect of biminne on vascular permeability of nasal mucosa in allergic rhinitis rats]. AB - OBJECTIVE: To investigate whether the effect of Biminne on allergic rhinitis (AR) was through improving vascular permeability of nasal mucosa. METHODS: Rat's model in Biminne-treated group and model group was induced by intraperitoneal injection of ovalbumin (OVA) and aluminum hydroxide gel suspension Biminne-treated rats were orally given Biminne suspension from the 8th day to the 17th day. On the 18th day, Evan's blue dye (EBD) in the nasal perfusate was detected to assess the vascular permeability. RESULTS: EBD concentration was higher in the model rats than that in the normal rats, and lower in the Biminne-treated rats than that in the model rats (both P < 0.01). CONCLUSION: Biminne could improve vascular permeability of nasal mucosa in sensitized rats, which may be the mechanism of its clinical effect on AR. PMID- 17121046 TI - [Effect of panax notoginseng saponins injection on brain edema in intracerebral hemorrhage rats]. AB - OBJECTIVE: To study the effect of panax notoginseng saponins (PNS) in treating hemorrhagic apoplexy at super-early stage in rats. METHODS: Rat model of hypertension with cerebral hemorrhage was induced by collagenase method. Sixty rats were randomly divided into 5 groups: the sham operated group, model group, PNS high, middle, and low dose group, 12 in each; 4 h after modeling, PNS or normal saline was intraperitoneally injected into the rats every 12 h, the total is 5 times. Contents of water, sodium and potassium ion in brain, and the diameter of hematoma in rats of different groups were measured 24 h and 72 h after modeling. RESULTS: Compared with the model rats, nerve defect symptoms aggravated, the contents of water and sodium ion in ipsilateral cortex and basal ganglia were significantly higher, the content of potassium ion was lower and the hematoma diameter was obviously less in the PNS-treated rats (all P < 0.05). CONCLUSION: PNS may worsen the brain edema and increase the nerve defect score when it was applied at the early stage of cerebral hemorrhage, but could promote the absorption of hematoma, indicating PNS should be used cautiously in treating patients with large amount of cerebral hemorrhage at super-early stage. PMID- 17121047 TI - [Effect of fulu baoxin oral liquid on lipid peroxidation, plasma nitric oxide and vascular endothelial growth factor in acute myocardial infarction rats]. AB - OBJECTIVE: To study the effect and mechanism of Fulu Baoxin Oral Liquid (FBOL) on lipid peroxidation (LPO), plasma nitric oxide (NO) and vascular endothelial growth factor (VEGF) in myocadial tissue of rats with acute myocardial infarction (AMI). METHODS: The contents of superoxide dismutase (SOD), malondialdehyde (MDA) and VEGF in myocardial tissue and the plasm NO level were determined by immunohistochemical method. RESULTS: Compared with those in AMI model rats, contents of SOD, plasma NO and myocardial VEGF were higher and MDA was lower obviously in the FBOL high dosage and coronary favorable medicine treated rats (P < 0.05 or P < 0.01). CONCLUSION: FBOL could increase SOD activity and lower MDA content in myocardial tissue, and up-regulate plasma NO and myocardial VEGF, which may be related to its curative effect on AMI. PMID- 17121048 TI - [Effects of acupuncture in Siguan points on man-machine counteraction in respiratory failure patients during mechanical ventilation]. AB - OBJECTIVE: To investigate the value of applying acupuncture in patients with respiratory failure Sixty (RF) during mechanical ventilation (MV) when there happened man-machine counteraction. METHODS: RF patients with nonsynchronous spontaneous breath (SB) during mechanical ventilation were divided into two groups randomly, 30 in each group. The control group treated with intravenous midazolam injection, and the treated group with acupuncture in Siguan points combined intravenous midazolam injection to maintain synchronized SB and MV, and to calm patients. The dose of midazolam used, the sedative effect as well as the complications occurred at different time points in the two groups were observed and compared. RESULTS: Compared with the control group, advantages such as smaller dose of midazolam used, lighter in deepness of sedation, and less complications were observed in the treated group (all P < 0.05). CONCLUSION: Acupuncture auxiliary Western medicine treatment has better effects than that of Western treatment alone in treating RF patients with man-machine counteraction during mechanical ventilation. PMID- 17121049 TI - [Effect of TCM syndrome-typing based therapy combined with cyclosporin in treating myelodysplastic syndrome]. AB - OBJECTIVE: To observe the therapeutic effect of TCM syndrome-typing based therapy (TCM-STBT) combined with cyclosporin on myelodysplastic syndrome (MDS) and investigate the correlation of efficacy with TCM syndrome type and MDS type. METHODS: Fifty-eight MDS patients were classified into refractory anemia (RA) and RA with excess blast (RAEB) by FAB standard and randomly divided into the cyclosporin group (the treated group) and the alltransretinoic acid group (the control group), and both were subjected to STBT additionally. The therapeutic effect and changes of peripheral hemogram were observed and compared. RESULTS: The total effective rate and the total remission rate was 73% and 33% in the treated group, higher than that in the control group (39% and 0) respectively. Hemoglobin level, white blood cell and blood platelet count increased significantly in the treated group after treatment (P < 0.05), while only hemoglobin level rose obviously in the control group (P < 0.05). The total effective rate and markedly effective rate in patients with qi-blood deficiency syndrome type were higher than those with other syndrome types. CONCLUSION: TCM STBT combined with cyclosporin has good efficacy in treating MDS and there is well correlation between the therapeutic efficacy and type of TCM syndrome or of MDS, indicating the STBT is necessory for TCM therapy. PMID- 17121050 TI - [Comparative study of the clinical effects of salvia miltiorrhiza injection and shengmai injection on chronic hepatitis B]. AB - OBJECTIVE: To observe the effect of Salvia miltiorrhiza Injection (SMI) and Shengmai Injection (SI) on liver function and fibrosis related indexes in patients with chronic hepatitis B. METHODS: Seventy-nine chronic hepatitis B patients were randomly divided into the SMI group (n=47) and the SI group (n=32), they were treated with SMI and SI respectively on the basis of conventional treatment. The therapeutic course was 35 days for both groups. The changes of main symptoms and physical signs were observed, and indexes of liver function and fibrosis including serum hyaluronidase, laminin, III type precollagen (PC-III) and IV type collagen (IV-C) were investigated before and after treatment. RESULTS: Symptoms, physical signs and liver functions were improved obviously in both SMI and SI groups, SI showed better effect than SMI (P < 0.05). The four liver fibrosis indexes declined significantly in the SMI group after treatment (P < 0.05), but no obvious change of those was found in the SI group (P > 0.05), showing significant difference between the two groups (P < 0.05). Conclusion SMI is effective in improving liver function and inhibiting liver fibrosis, and SI has even better effect in improving liver function than SMI, though it shows no anti-liver fibrosis effect. PMID- 17121051 TI - [Effects of pulmonary arterial perfusion with shenqi fuzheng injection on lung injury during cardiopulmonary bypass]. AB - OBJECTIVE: To observe the effect of pulmonary arterial perfusion with Shenqi Fuzheng Injection (SFI)on lung injury during cardiopulmonary bypass (CPB). METHODS: Twenty-two patients with cardiac valvular disease and pulmonary hypertension were randomly divided into the control and the SFI group equally. SFI mixed pure oxygenated blood to the SFI group, and oxygenated blood alone to the control group was perfused via pulmonary artery during CPB. Plasma malondialdehyde (MDA), pulmonary vascular resistance (PVR), ratio of leucocyte counts in venous and arterial blood, and time of mechanical ventilation applied were measured before and at the end of CPB, and 6th, 24th hours after CPB. RESULTS: After treatment, MDA content and PVR were significantly higher than those before CPB (P < 0.05), and reduced to normal level 24 h after CPB in both groups, but the peak levels were lower in the SFI group than those in the control group (P < 0.05). The leucocyte counts ratio in venous and arterial blood were significantly higher at the end of CPB and 6 h later than those before CPB in both groups (P < 0.05), but the increment were lower in the SFI group than those in the control group (P < 0.05). Furthermore, the applying mechanical ventilation time in the SFI group was 16.1 +/- 5.5 h, significantly shorter than that in the control group (29.1 +/- 6.9 h, P < 0.01). CONCLUSION: Pulmonary arterial perfusion with SFI could alleviate the CPB induced lung injury. PMID- 17121052 TI - [Diagnostic standard of integrative Chinese and Western medicine for cerebral infarction and hemorrhage (try out)]. PMID- 17121053 TI - [Alienation and adaptation in English translation of traditional Chinese medicinal literature]. AB - Alienation and adaptation are two of the principles and methods in translation, each possessing their own values. Alienation should be applied in translating linguistic content in order to transfer the imbedded cultural messages trustfully; while the principle of adaptation should be followed in linguistic structure translation due to the different thinking patterns between Chinese and English which results in a great linguistic structure difference. Therefore, the translator must express the original meaning trustfully, on the other hand, to make the Chinese version more smooth, linguistic structure should be transformed to conform to the thinking habit of the readers. In brief,alienation and adaptation should complement each other in translation to make it a bridge connecting the different cultures. PMID- 17121054 TI - [Criticism of "English translation of common terms in Chinese medicine"]. AB - This paper offers criticism of "English Translation of Common Terms in Chinese Medicine" (2004) by Professor XIE Zhu-fan. Criticisms of specific term choices are listed under the following headings: (1)Poor Term Choices;(2) Poor Grammar and Style; (3) One Term with Multiple Translations (Inconsistency); (4) Multiple Terms with one Translation; (5) Overuse of Pinyin; (6) Overuse of Modern Medical Terms. PMID- 17121055 TI - [Progress of research on effects of ginsenoside Rg1 in promoting capability of learning and memory]. AB - It is testified by long-standing traditional Chinese medicine clinical practice that ginseng was effective in treating dementia and promoting capability of learning and memory, for which ginsenoside Rg1 has been proved the main effective ingredient. Recently many researches have been carried out on the mechanism and action links of ginsenoside Rg1, and illustrated that it could exert the anti dementia and nootropic effects through intervening multiple targets and links, thus to provide a theoretical basis for bettering the clinical use of ginsenoside Rg1. PMID- 17121056 TI - Observations on the ongoing shortage of academic dermatologists. PMID- 17121057 TI - What's eating you? Common striped bark scorpion (Centruroides vittatus). PMID- 17121058 TI - Photo quiz. What is your diagnosis? Confluent and reticulate papillomatosis (Gougerot-Carteaud syndrome). PMID- 17121059 TI - Atopic dermatitis in children, part 1: epidemiology, clinical features, and complications. AB - Atopic dermatitis (AD), also known as eczema, is a chronic skin condition, characterized by itch (pruritus) and dryness (xerosis). AD lesions appear as pruritic red plaques that ooze when scratched. Children with AD are excessively sensitive to irritants such as scented products and dust due to their impaired skin barrier and skin immune responses. AD is among the most common disorders of childhood and its incidence is increasing. AD is an all-encompassing disease that causes sleep disturbances in the affected child, disrupting the entire household. Patients with AD also are prone to bacterial overgrowth, impetigo, and extensive viral infections. Consequently, familiarity with the most recent literature is of utmost importance so that dermatologists and pediatricians can appropriately manage their patients. PMID- 17121060 TI - Lymphocutaneous nocardiosis from commercially treated lumber: a case report. AB - GOAL: To understand nocardiaI infections to better manage patients with the condition. OBJECTIVES: 1. Identify the organisms causing nocardial infections in humans. 2. Describe the presenting symptoms of nocardial infections. 3. Explain the treatment of nocardial infections. PMID- 17121061 TI - Fatal sporotrichosis. AB - Disseminated sporotrichosis is a serious fungal infection caused by the soil inhabitant Sporothrix schenckii. It is seen in immunocompromised patients, with a substantial number of recent cases involving patients with acquired immunodeficiency syndrome (AIDS). However, individuals with other conditions that affect the immune system also are at increased risk. We report a case of fatal disseminated sporotrichosis in a patient with liver disease and a diagnosis of a granulomatous condition presumed to be sarcoidosis; the patient was receiving systemic corticosteroid therapy. The various presentations of S schenckii infection, the risk of disseminated disease in immunocompromised hosts, and the importance of making accurate histologic diagnoses are reviewed. PMID- 17121062 TI - Tacrolimus ointment 0.1% for the treatment of peristomal skin disease: 3 case reports. AB - Patients with stomas face a variety of problems, such as skin breakdown or ulceration at the peristomal site, that can complicate care. Topical steroids are frequently used to treat various inflammatory conditions that affect peristomal skin with good results, but chronic use can lead to undesirable side effects. Tacrolimus ointment 0.1%, a nonsteroidal immunosuppressant, could offer a more favorable alternative to topical steroids. We present 3 cases of peristomal skin disease that were successfully treated with tacrolimus ointment 0.1%. PMID- 17121063 TI - Inflammatory linear verrucous epidermal nevus: a case report and short review of the literature. AB - GOAL: To understand inflammatory linear verrucous epidermal nevus (ILVEN) to better manage patients with the condition. OBJECTIVES: Upon completion of this activity, dermatologists and general practitioners should be able to: 1. Describe the presenting characteristics of ILVEN. 2. Explain the differential diagnosis of ILVEN. 3: Discuss the treatment options for ILVEN. PMID- 17121064 TI - The safety and efficacy of sertaconazole nitrate cream 2% for tinea pedis. AB - To determine the safety and efficacy of topical sertaconazole nitrate cream 2% in the treatment of tinea pedis, 2 randomized, multicenter, double-blinded, parallel group, vehicle-controlled studies were conducted. A total of 588 subjects were enrolled, and 383 subjects were randomized to treatment with sertaconazole or vehicle applied twice daily for 4 weeks. Improvements in symptoms were noted at week 1 in the active treatment group. At week 4, mycologic cure was seen in 70.3% of sertaconazole-treated subjects and 36.7% of vehicle-treated subjects (P<.0001). At week 6, 46.7% of sertaconazole-treated subjects had successful treatment outcomes compared with 14.9% of vehicle-treated subjects (P<.0001). No serious adverse events were reported, and rates of cutaneous adverse events were comparable between treatment groups. In conclusion, sertaconazole nitrate cream 2% was well-tolerated, offered rapid relief of symptoms, and achieved high rates of mycologic cure. The stability of the mycologic cure rates through weeks 5 and 6 (2 weeks after cessation of therapy) indicate that sertaconazole protects against reinfection. PMID- 17121066 TI - [Prospects in the treatment of patients with chronic renal failure?]. PMID- 17121065 TI - Facilitating facial retinization through barrier improvement. AB - The utility of topical tretinoin as a treatment for improving the appearance of photodamaged skin is limited by irritation that occurs during the early phases of facial retinization. The observed side effects are consistent with stratum corneum barrier compromise. This paired double-blinded study was conducted to determine if preconditioning the skin with a barrier-enhancing cosmetic facial moisturizer before beginning tretinoin therapy and continuing moisturizer application during therapy would mitigate these side effects. Women with facial photodamage were recruited and randomly assigned to apply one cosmetic moisturizer to one side of the face and the other cosmetic moisturizer to the other side of the face twice daily for 10 weeks. One moisturizer contained a mixture of vitamins (niacinamide, panthenol, and tocopheryl acetate) to enhance stratum corneum barrier function, and the other moisturizer contained similar moisturizing ingredients but no vitamins. Daily full-face treatment with tretinoin cream 0.025% commenced 2 weeks into the study. Subjects' facial skin condition was monitored via investigator assessments, instrumental measurements, and subject self-assessments. The results show that improving stratum corneum barrier function before beginning topical tretinoin therapy and continuing use of a barrier-enhancing cosmetic moisturizer during therapy facilitates the early phase of facial retinization and augments the treatment response. PMID- 17121067 TI - [Interstitial microdialysis in the clinical and experimental medicine]. AB - Interstitial microdialysis is a minimally invasive method that allows biochemical monitoring of endogenous and exogenous substances in extracellular tissue space. The method is based on sampling of soluble molecules from the interstitial space fluid by means of a semipermeable membrane. Microdialysis has been applied in primary and clinical research of metabolic changes and the blood flow in tissue interstitium and pharmacokinetic drug studies. Results obtained by microdialysis are gradually finding their place in clinical practice as well. This paper is aimed at presenting to the reader new technique of monitoring interstitial metabolism, its advantages, drawbacks and relevance for medicine. PMID- 17121068 TI - [Glomerular filtration rate prediction formulas in obese persons with chronic renal impairment]. AB - BACKGROUND: The glomerular filtration rate (GFR) could be estimated on the basis of serum creatinine concentration (Scr) and some simple variables and demographic data. In clinical practice the most used methods for GFR estimation are Cockcroft Gault (CG) formula and abbreviated MDRD equation (MDRD abr). The aim of this study was to evaluate how far obesity could affect GFR estimation based on the above formulas. METHODS AND RESULTS: In 291 patients with chronic renal impairment (S(cr) 45-489 micromol/l) GFR was examined on the basis of renal inulin clearance (C(in)) and estimated using MDRD abr and CG (without correction for body surface area-BSA and CG corrected for BSA) (CGkorig). The group of nonobese patients (A) consisted of 229 patients (BMI <30 kg/m2) and the group of obese patients (B) consisted of 62 patients (BMI 30 kg/m2). The values of r (r2) for MDRD abr, CG and CG(korig) in group A of patients was as follows: 0.893 (0.797), 0.810 (0.651), 0.853 (0.727) and 0.853 (0.727). In obese patients (group B) the corresponding values were as follows: 68.3% (82.6%), 28.6% (39.7%) and 46% (61.9%). Predicted GFR within 30% and 50% of C(in) (in brackets), CG and CG(korig) (for BSA) were for group A: 70.2% (87.3%), 50.4% (67.1%) and 55.7% (75%) and for group B: 68.3% (82.6%), 28.6% (39.7%) and 46% (61.9%). The ratio MDRD abr/Cin did not correlate with BMI. A weak, but significant correlation was found between BMI and CCcorig/Cin ratio (r=0.22, p<0.05). CONCLUSIONS: The obtained results suggest that estimation of GFR based on MDRD abr is not influenced by obesity. Estimation of GFR based on CG formula is significantly affected by obesity. A weak but significant relation could by found between CGkorig/Cin and BMI. PMID- 17121069 TI - [Effects of specific cycloxygenase-2 inhibition on the renal functions of ederly patients with renal function impairment]. AB - BACKGROUND: Elderly patients suffering from nociceptive pain of locomotive organs and concomitantly from renal impairment represent a target population for painkilling drugs. That is why they are predisposed to nephrotoxic effects non steroidal anti-inflammatory drugs. The aim of our study was to evaluate cycloxygenase-2 (COX-2) inhibition effect on renal function in elderly with moderate impairment of renal function. METHODS AND RESULTS: Based on 24-h urine collection we assessed creatinine clearance (C(Cr), fractional excretion of sodium (FE(Na)), potassium (FE(K)), chloride (FE(Cl)), osmotic active solutes (FE(OSM)) and 24h urinary excretion of prostaglandin PGE2 and PGF(2 alpha). Under conditions of sub-maximal water load fractional excretion of electrolytes, inulin clearance (C(in)), serum cystatin C (S(cyst)) were assessed. In addition basal and stimulated plasma renin activity (PRA) and plasma aldosteron (P(aldo)) were examined. Using comparison of parameters before and at the end of 7-days rofecoxib treatment we found out C(in) 0,82 +/- 0,34 vs 0,74 +/- 0,18 ml/s/l,73 m2, FE(Na) 1,0 +/- 0,3 vs 1,2 +/- 0.4 (p=0,02), FE(OSM) 2.9 +/- 0,7 vs 3,7 +/- 1,2% (p=0,03), U(PGE2 alpha),V 663 +/- 528 vs 414 +/- 195 (p=0,059), U(PGD2) V (559 +/- 625) vs 205 +/- 174 eta g/24h (p=0,02), stimulated PRA 0.94 +/- 0,73 vs 0,4 +/- 0,27 +/- pg/l/h (p=0,019), P(aldo) 104,56 +/- 50,15 vs 56,94 +/- 27,08 eta g/l/h (p=0,008). CONCLUSIONS: Short-term COX-2 inhibition in patients with moderate renal impairment was associated with significant decrease of tubular transport of sodium, without changing GFR and water excretion. PMID- 17121070 TI - [Motor skills testing in patients with chronic renal failure]. AB - BACKGROUND: During chronic renal failure, number of complex metabolic and endocrinal changes in organism can develop, which in consequence lead to an inception of associate complications and in principle change the activity of all organ systems. Functional capacity of the chronically dialysed patients is limited, muscle strength is decreased, joint mobility is limited and the muscular imbalance is frequently found. According to the recent literature, an acceptable choice of exercise activity could contribute to prevention of disturbances and improve the patient's status. An essential part of each exercise program for these patients is an acceptable motor tests battery (1, 2, 3, 4, 5, 6 and 7). The aim of our study was to choose an acceptable motor tests battery, which should target such components of motor performance, whose certain rate is necessary for self-sufficiency keeping and perform activities of daily living. METHODS AND RESULTS: We observed mixed group of 23 patients. For evaluation of the functional condition we used the "Senior Fitness Test Manual" (8), which measures these physical attributes: muscle strength, physical endurance, flexibility, agility and balance. The input results we compared with population standard specification used in the test battery. Results of the tests showed that the group of patients in comparison with the population standard specification have reached subnormal and risk performances, especially in tests requiring for its implementation muscular strength of lower extremities and physical efficiency. Normal and above normal performances we observed in patients that were physically active before and during regular dialysis treatment. CONCLUSIONS: After the evaluation of result, we consider the selected battery of motor tests as an acceptable choice for motor skills testing in renal dialysed patients all age categories. PMID- 17121071 TI - [Ischemic disease of the lower extremities versus depression symptoms and quality of life]. AB - BACKGROUND: The study has three main aims: 1. it evaluates occurrence of depression symptoms in patients with peripheral arterial occlusive disease (PAOD), 2. it evaluates global quality of life (QoL) in patients with PAOD and 3. it evaluates effect of age and stage of PAD on seriousness of depression symptoms and on the QoL in patients with PAOD. PATIENTS AND METHOD: the study is local, prospective and cross-sectional. It was carried out at the 2nd Internal Clinic of University Hospital in Hradec Kralove. Data were obtained during the year 2006. METHODS AND RESULTS: The total number of respondents with PAOD was 42 (28 male, 14 female). The average age of all respondents with PAOD was 65.4 years and age range was 45-79 years. The evaluation of occurrence of depression symptoms in patients with PAOD was performed by means of self-assessment Zung-SDS and evaluation of QoL in patients with PAOD was performed by means of Czech version of international generic European Quality of Life Questionnaire-EQ-5D Version. Statistical significance was determined by means of analysis of variance. Main The mean SDS index certifies the presence of signs of minimum or light depression in patients with PAOD. The QoL in patients with PAOD is on the low level. The above-mentioned aspects proved statistically significant dependence of QoL in patients with PAOD on depression (p<0.001), on age (p<0.01) and on stage of PAOD (p<0,01). We proved statistically significant dependence of depression in patients with PAOD on age (p<0.01) and on stage of PAOD (p<0.01). CONCLUSIONS: The results show the existence of the association between PAOD and depression symptoms with its negative effect on QoL in patients. PMID- 17121072 TI - [Photodynamic therapy with visudyne in treatment of wet form of age-related macular degeneration--two years results]. AB - BACKGROUND: Photodynamic therapy with Visudyne represents a new method in treatment of the wet form of age-related macular degeneration. METHODS AND RESULTS: Effectiveness of photodynamic therapy with Visudyne was confirmed at our department in the group of 49 patients (13 men, 36 women) of mean age 72.7 years with predominantly classical and occult choroidal neovascular membrane in subfoveal localization in wet form of age-related macular degeneration. The observation period in all patients in this group was 24 month. During the observation period, 27 patients with predominantly classical choroidal neovascular membrane underwent 1 to 5 treatments (mean 1.8). Before the treatment the average best corrected visual acuity was 0.696 +/- 0.20 logMAR. At the end of observation period the average best corrected visual acuity was 0.985 +/- 0.39 logMAR. Best corrected visual acuity dropped by 2.88 lines of ETDRS (early treatment diabetic retinopathy study) visual charts. 22 patients with the genuine occult choroidal neovascular membrane underwent during the observation period 1 to 3 treatments (mean 1.5). Average best corrected visual acuity before the treatment was 0.755 +/- 0.25 logMAR. At the end of observation period the average best corrected visual acuity was 0.909 +/- 0.42 logMAR. We found out the decrease of average best corrected visual acuity by 1.55 lines of ETDRS charts. CONCLUSIONS: Decrease of the average best corrected visual acuity less than 3 lines at ETDRS charts is considered as stabilisation of the finding. In our group this goal was achieved in 2/3 of patients. PMID- 17121073 TI - [Results of photodynamic treatment of choroidal neovascular membrane in age related macular degeneration]. PMID- 17121074 TI - [Role of percutaneous vertebroplasty and kyphoplasty in the treatment of oncology disorders of the spine]. AB - BACKGROUND: The aim of the study is to present results of a prospective uncontrolled clinical study. Percutaneous vertebroplasty or kyphoplasty are minimally invasive methods based on polymethylmethacrylate (PMMA) bone cement application into the damaged vertebra. This leads to decrease of the pain and vertebral body stabilisation. Oncology disorders of the spine are relatively common, having a wide alternative of various methods of treatment. Patients, according to their findings and indication criteria, are treated surgically or conservatively, oncological treatment is usually based on radiotherapy. Authors discuss the role of these invasive procedures in the treating algorithm of patients with spinal metasthases and multiple myeloma. METHODS AND RESULTS: From September 2003 to December 2005, 21 percutaneous vertebroplasties in 14 patients, mean age 68.7 (47-80) year, were performed in our department. During one treatment session 1-2 vertebrae (total of 21 vertebrae) in level Th9 - L5 were treated. Vertebroplasties and kyphoplasty were performed under fluoroscopy guidance. Transpedicular acces was used. Totally, 3 asymptomatic complications were proved. As first, a bone cement leaked paravertebrally during L5 body treatment, as second, a bone cement leaked into paravertebral veins, and as third, a bone cement leaked into the intervertebral space. Visual analog scale (VAS) was 8.9 points before procedure, 1.9 point 3 months after procedure and 2.6 points 6 months after procedure. We did not prove a symptomatic or total complication. CONCLUSIONS: According to our experience, percutaneous vertebroplasty is an effective alternative treatment of painful oncologic spine disease. PMID- 17121075 TI - [Cardiac myxoma in the left atrium resulting in syncopes]. AB - A case study of a 74-year-old woman with cardiac myxoma in the left atrium resulting in syncopes is discussed in this article. PMID- 17121076 TI - [What did the study of gene polymorphisms contribute to practise?]. PMID- 17121077 TI - [Development of pharmacy in the Czech Republic]. PMID- 17121078 TI - Effect of probe-site mismatches on detection of virulent Newcastle disease viruses using a fusion-gene real-time reverse transcription polymerase chain reaction test. AB - Virulent forms of Newcastle disease virus (NDV) are a major concern for poultry producers around the world and the rapid diagnosis of an outbreak is crucial to any control program. A validated real-time reverse transcription-PCR test (fusion test) directed at the fusion-cleavage site of NDV was developed to differentiate virulent Newcastle disease virus strains from those of low virulence, however one virulent isolate, Dove/Italy/2736/2000, escaped detection during the initial evaluation of the test. The objectives of this study were to determine how this isolate differed from other detectable isolates, to identify other isolates that may fail detection, and to characterize the effect of specific probe-site mutations on the fusion test at a range of annealing temperatures. Using a virulent NDV isolate (Game fowl/US(CA)/2002) as a backbone that has 100% identity to the fusion-test probe, specific changes were made to the fusion-test probe site to reflect the unique mismatches found in Dove/Italy/2736/2000 and other selected regions of the probe. Mutated clones with mismatches unique to Dove/Italy/2736/2000 at positions 6, 13, and 14 were not detected until annealing temperatures were lowered to 50 degrees C. Those detected at 58 degrees C contained 1-2 mismatches (position 1 and 6, 13 and 14, or 14 only) although increased cycle threshold values compared to the parent clone indicated decreased sensitivity. Data from this study predicts that the fusion test may fail to detect some viruses among lineage 4b and potential solutions to identify this subset of viruses include lowering the annealing temperature or modifying the probe. PMID- 17121079 TI - A testing scheme for the detection of Mycobacterium avium subsp. paratuberculosis in bovine feces utilizing the ESP para-JEM liquid culture system. AB - A testing scheme for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) in broth cultures of bovine fecal samples carried out in ESP para-JEM System was evaluated. The scheme included acid-fast staining (on signal-positive and signal-negative samples), and confirmation by PCR for 2 MAP-specific targets and subculture of all acid-fast positive PCR-negative samples. Two hundred and fifty bovine fecal samples were evaluated for the presence of MAP using this scheme. Thirty-seven (15%) of 250 fecal samples had a positive culture result when the proposed testing scheme was used, compared to 14 (6%) positive results when using the standard ESP para-JEM protocol (requiring samples to have a positive signal from the system, a positive acid-fast stain, and a positive IS900 PCR result), and 20 (8%) positives when conventional culture was performed on Herrold egg yolk (HEY) media. A preliminary comparison of real-time and conventional PCR on DNA extracted from 15 MAP-positive broth cultures by 3 different protocols suggested that conventional PCR may be a better choice for the confirmation of the presence of MAP in the liquid cultures than real-time PCR. PMID- 17121080 TI - Tissue distribution of psittacid herpesviruses in latently infected parrots, repeated sampling of latently infected parrots and prevalence of latency in parrots submitted for necropsy. AB - Psittacid herpesvirus-1 (PsHV-1) is the cause of an acute fatal disease in parrots and is implicated as the cause of papillomatous lesions of the digestive tract. Not all infections cause disease and some parrots are infected asymptomatically. Latently infected parrots are potential sources for virus dissemination. Tissues from parrots that died spontaneously with a history of coming from flocks where a PsHV-1 outbreak had occurred were examined for PsHV-1 DNA. Fourteen of 16 parrots examined were infected with at least 1 variant of PsHV-1; of these 13 (93%) had viral DNA in either or both the oral and cloacal mucosa, suggesting that most latently infected parrots could be detected by sampling these sites. Nine of 9 parrots shown to be infected 5 years prior to this study were positive again on repeat sampling and were infected with the same virus genotype. Opportunistic sampling of parrots submitted for diagnostic necropsy indicated that the prevalence of PsHV-1 in parrots in the sampled population was approximately 9.3%. PsHV-1 genotypes 1, 2, and 3 were found in these birds, but genotype 4 was not. Six necropsy specimens were found to be infected with two PsHV-1 genotypes and it was concluded that infection with one serotype did not protect against infection with another. Psittacid herpesvirus 2 (PsHV-2) was identified in 4 African grey parrots and a blue and gold macaw. Prior to this study PsHV-2 had only been found in African grey parrots. PMID- 17121081 TI - Development and evaluation of an indirect enzyme-linked immunosorbent assay for detection of foot-and-mouth disease virus nonstructural protein antibody using a chemically synthesized 2B peptide as antigen. AB - Forty peptides were synthesized corresponding to hydrophilic clusters of amino acids within the sequences of foot-and-mouth disease virus (FMDV) nonstructural proteins (NSP). Six peptides were studied in more detail and the most promising, a 2B peptide, was evaluated in enzyme-linked immunosorbent assay (ELISA) using sera from naive, vaccinated, and vaccinated-and-challenged cattle as well as bovine sera from field outbreaks. The performance of the new NSP peptide ELISA was compared to that of 4 commercial NSP ELISA kits. Antibody to 2B was detectable from the end of the first week to the second week after infection in most of the nonvaccinated animals and by the second to third week in vaccinated and-challenged animals. The sensitivity of the 2B peptide ELISA was comparable to the 3ABC Ceditest (Ceditest FMDV-NS, Cedi Diagnostics B.V.; Chung et al., 2002). With some modification and further validation, this 2B test could be useful as a screening or conformational NSP test in postvaccination surveillance for FMD. PMID- 17121082 TI - Detection of PrP(CWD) in postmortem rectal lymphoid tissues in Rocky Mountain elk (Cervus elaphus nelsoni) infected with chronic wasting disease. AB - Preclinical diagnostic tests for transmissible spongiform encephalopathies have been described for mule deer (Odocoileus hemionus), using biopsy tissues of palatine tonsil, and for sheep, using lymphoid tissues from palatine tonsil, third eyelid, and rectal mucosa. The utility of examining the rectal mucosal lymphoid tissues to detect chronic wasting disease (CWD) was investigated in Rocky Mountain elk (Cervus elaphus nelsoni), a species for which there is not a live-animal diagnostic test. Postmortem rectal mucosal sections were examined from 308 elk from two privately owned herds that were depopulated. The results of the postmortem rectal mucosal sections were compared to immunohistochemical staining of the brainstem, retropharyngeal lymph nodes, and palatine tonsil. Seven elk were found positive using the brainstem (dorsal motor nucleus of the vagus nerve), retropharyngeal lymph nodes, and palatine tonsil. Six of these elk were also found positive using postmortem rectal mucosal sections. The remaining 301 elk in which CWD-associated abnormal isoform of the prion protein (PrP(CWD)) was not detected in the brainstem and cranial lymphoid tissues were also found to be free of PrP(CWD) when postmortem rectal mucosal sections were examined. The use of rectal mucosal lymphoid tissues may be suitable for a live-animal diagnostic test as part of an integrated management strategy to limit CWD in elk. PMID- 17121083 TI - Transmission of chronic wasting disease of mule deer to Suffolk sheep following intracerebral inoculation. AB - To determine the transmissibility of chronic wasting disease (CWD) to sheep, 8 Suffolk lambs of various prion protein genotypes (4 ARQ/ARR, 3 ARQ/ARQ, 1 ARQ/VRQ at codons 136, 154, and 171, respectively) were inoculated intracerebrally with brain suspension from mule deer with CWD (CWDmd). Two other lambs were kept as noninoculated controls. Within 36 months postinoculation (MPI), 2 inoculated animals became sick and were euthanized. Only 1 sheep (euthanized at 35 MPI) showed clinical signs that were consistent with those described for scrapie. Microscopic lesions of spongiform encephalopathy (SE) were only seen in this sheep, and its tissues were determined to be positive for the abnormal prion protein (PrP(res)) by immunohistochemistry and Western blot. Three other inoculated sheep were euthanized (36 to 60 MPI) because of conditions unrelated to TSE. The 3 remaining inoculated sheep and the 2 control sheep did not have clinical signs of disease at the termination of the study (72 MPI) and were euthanized. Of the 3 remaining inoculated sheep, 1 was found to have SE, and its tissues were positive for PrP(res). The sheep with clinical prion disease (euthanized at 35 MPI) was of the heterozygous genotype (ARQ/VRQ), and the sheep with subclinical disease (euthanized at 72 MPH) was of the homozygous ARQ/ARQ genotype. These findings demonstrate that transmission of the CWDmd agent to sheep via the intracerebral route is possible. Interestingly, the host genotype may play a notable part in successful transmission and incubation period of CWDmd. PMID- 17121084 TI - Modest copper supplementation blocks molybdenosis in cattle. AB - It is widely accepted that ratios of dietary copper (Cu) to molybdenum (Mo) lower than 10:1 may produce molybdenosis in cattle, especially if sulfur concentrations are more than 3,000 ppm. Some authorities suggest that dietary Mo concentrations greater than 10 ppm are hazardous to cattle regardless of Cu concentration, but anecdotal reports suggest that this may not be the case. The original purpose of the experiment described in this report was to investigate whether supranutritional supplemental Cu could protect cattle against relatively high dietary Mo. Pregnant cows were grazed on 1 of 3 pastures: 1 with only background Mo, 1 with an average of 13 ppm Mo, and 1 that averaged 230 ppm Mo. Half the cows on the Mo pastures were supplemented with 17 ppm dietary Cu, the other half with the dietary supplement plus Cu boluses. Molybdenum effects were anticipated in the groups supplemented with 17 ppm Cu; however, despite increased tissue concentrations of Mo, only the 230 ppm Mo/17 ppm Cu group exhibited any effects. Moderate Cu supplementation permitted cows to graze a site heavily contaminated with Mo with no adverse effects on general health or reproduction. PMID- 17121085 TI - Aplasia cutis congenita (epitheliogenesis imperfecta) in swine: observations from a large breeding herd. AB - Epitheliogenesis imperfecta has been reported in several animal species, and its inheritance is suspected to be autosomal recessive. This term has been used to describe two different diseases, namely epidermolysis bullosa and aplasia cutis congenita, which are both grossly characterized by an absence of epidermis or mucosal epithelium and are most frequently reported on the distal limbs and oral cavity. Epitheliogenesis imperfecta has been described in swine, but the literature on the subject is scarce. To better characterize this condition, 70 piglets with congenital skin defects macroscopically compatible with epitheliogenesis imperfecta were examined. In all but 1 case, only 1 piglet per litter was affected. Of the affected piglets, 65 (93%) were male, suggesting a sex-related problem. More than half of the piglets had multiple skin lesions. All defects were located on the caudal half of the body, and none was found in the oral cavity. Most lesions were characterized by an absence of epidermis and part of the dermis and adnexae. Adnexal dysplasia was also observed at several sites, both with and without epitheliogenesis imperfecta, suggesting a developmental problem. Fluid-filled, congenital subcutaneous bullae were noted grossly on 7 piglets; their relationship, if any, with epitheliogenesis imperfecta remains unknown. As the term epitheliogenesis imperfecta has been used in cases of epidermolysis bullosa, the term aplasia cutis congenita seems to be more appropriate to describe these lesions in swine. PMID- 17121086 TI - Freezing complete polymerase chain reaction master mix reagents for routine molecular diagnostics. AB - The potential of storing complete frozen real-time polymerase chain reactions (PCRs) and real-time reverse transcription PCRs (RT-PCRs), which require only thawing and the addition of template nucleic acid before PCR cycling, was examined. Master mixes containing all necessary reagents at working concentration (except template nucleic acid) were aliquoted into single-reaction volumes and stored at -70 degrees C for periods of up to 8 months. Reactions were removed from storage and nucleic acid template was added and amplified using different real-time PCR instruments. Threshold cycle values were used to monitor changes in assay performance after storage. Results for hybridization probe and TaqMan probe assays showed that freezing complete real-time PCR and RT-PCR reaction mixes was possible without deterioration in assay performance. This approach has advantages for routine molecular diagnostics in areas such as convenience, test consistency, quality control, and ease of use by nonspecialist staff. PMID- 17121087 TI - Comparison of direct fluorescent antibody staining and real-time polymerase chain reaction for the detection of Borrelia burgdorferi in Ixodes scapularis ticks. AB - Borrelia burgdorferi, the agent responsible for causing Lyme disease in humans and animals, is transmitted via the bite of infected Ixodes spp. ticks. Ticks removed from humans and animals are routinely tested by diagnostic laboratories to determine if they are infected with these bacteria. The objective of this study was to compare the efficacy of 2 commonly used methods, direct fluorescent antibody staining and real-time polymerase chain reaction (PCR), for the detection of B. burgdorferi in Ixodes scapularis ticks. One hundred and twenty seven adult I. scapularis ticks collected in Connecticut, a Lyme disease endemic area, were tested, and results were compared. Results showed 24.8% ticks tested positive for Borrelia spp. by fluorescent antibody testing and 32.5% ticks were positive for B. burgdorferi by real-time PCR testing. When ticks were grouped into categories by level of engorgement (unengorged, partially engorged, and fully engorged), 95% of unengorged ticks, 90.5% of partially engorged, and 86.8% of engorged ticks tested were in agreement. Ten of the 127 ticks examined were too dehydrated to be tested by the fluorescent antibody technique; half of these tested positive by PCR. Real-time PCR appears to be the better of these 2 methods for the diagnosis of this bacterial infection in I. scapularis ticks. PMID- 17121088 TI - Comparison of three DNA preparation methods for real-time polymerase chain reaction confirmation of Mycobacterium avium subsp. paratuberculosis growth in an automated broth culture system. AB - Three methods of harvesting DNA from broth culture tubes for quantitative real time polymerase chain reaction (qrtPCR) confirmation of Mycobacterium avium subsp. paratuberculosis (MAP) were evaluated. A commercial DNA extraction kit, the boil method (boiling for 5 minutes), or direct addition of broth culture media to the PCR reaction mix were tested. Samples were evaluated at 8 or 11 days of incubation and at the time of instrument-signal culture-positive. In total, when tested at time to instrument signal positive, 10/10 (100%) of samples extracted by the commercial method were positive on qrtPCR, whereas 9/10 (90%) were positive after the boil method, and 6/10 (60%) were positive after the direct method. Increased volumes of egg-yolk emulsion added to the culture tubes prolonged the number of cycles to threshold positive for the samples that were not subjected to commercial extraction or boiling. Samples were not reliably positive when tested at 8 or 11 days of incubation. The boil method appears to represent a reasonable time- and money-saving method to harvest DNA for qrtPCR confirmation of MAP in broth culture at time to instrument signal positive. PMID- 17121089 TI - Cyclooxygenase-2 expression in feline pancreatic adenocarcinomas. AB - Cyclooxygenase-2 (COX-II) is an inducible enzyme that is responsible for the production of prostaglandin E2 (PGE2), which is often upregulated in neoplastic conditions. Expression of COX-II is documented in the majority of human pancreatic adenocarcinomas and in many epithelial neoplasms in humans and animals. The purpose of this study was to assess a series of feline pancreatic adenocarcinomas for the expression of COX-II. Eight feline pancreatic adenocarcinomas (5 poorly differentiated ductular variants and 3 well differentiated acinar variants) were included. Immunohistochemical staining showed that COX-II was expressed in 2 (both poorly differentiated ductular variants) of the 8 neoplasms (25%). Approximately 10% of the epithelial cells from these 2 neoplasms expressed intense cytoplasmic staining. However, because feline pancreatic adenocarcinoma does not appear to consistently express COX-II, it is not a useful prognostic indicator for this group of feline neoplasms. In addition, COX-II inhibitors are not likely to be effective therapeutics for cats with this neoplasm. PMID- 17121090 TI - Isolation of Vibrio cholerae from the brain of a feedlot heifer with meningoencephalitis. AB - A 700-pound, 9-month-old Angus heifer from a feedlot presented with acute neurologic signs, characterized by circling, posterior weakness, and nonresponsiveness, followed by death. Histologically, the frontal lobe and the thalamus contained multiple foci of liquefaction that contained numerous degenerative neutrophils and foamy macrophages. Some of these foci were centered on blood vessels that contained fibrin thrombi and exhibited varying degrees of fibrinoid necrosis of the vessel wall. There was adjacent axonal degeneration and neuronal necrosis characterized by pronounced cytoplasmic eosinophilia, peripheralization of the nuclei, and loss of Nissl substance. Aerobic culture of the brain yielded moderate growth of Vibrio species, which was determined to be Vibrio cholerae by polymerase chain reaction analysis of a 438-base pair fragment of the 16 S ribosomal RNA gene. V. cholerae are motile, gram-negative, curved rod shaped bacteria. Some strains of V. cholerae are important food- and water-borne bacterial pathogens that produce an often fatal diarrhea in humans. This is the first known case report of V. cholerae meningoencephalitis and cerebral abscessation in a bovine. PMID- 17121091 TI - Screening petting zoo animals for the presence of potentially pathogenic Escherichia coli. AB - Several outbreaks of Escherichia coli O157 have been reported in petting zoos, resulting in hospitalization of many children. At present, no standard procedure has been adopted to monitor the presence of enterohemorrhagic E. coli (EHEC) or Shiga-toxin-producing E. coli (STEC) in petting zoo animals. Direct detection of these strains from rectal swabs of animals in petting zoos was developed and obviated the need to culture the organisms. DNA extracted from bacteria in the swabs was tested for the presence of wecA gene specific for E. coli by polymerase chain reaction (PCR). The wecA positive samples were further tested for Shiga toxin genes stxl and stx2, and the intimin eae by multiplex PCR and for the presence of O157 and H7. Swabs (n=104) from 15 animal species in a petting zoo were tested; 7 goats and 3 cows were found to carry STEC. The method is rapid and convenient for monitoring potentially pathogenic E. coli in petting zoo animals. PMID- 17121092 TI - A survey of bovine viral diarrhea virus testing in diagnostic laboratories in the United States from 2004 to 2005. AB - Bovine viral diarrhea virus (BVDV) has a great economic impact on the United States cattle industry. The Academy of Veterinary Consultants, the American Association of Bovine Practitioners, and the National Cattlemen's Beef Association have called for the goal of BVDV control and eventual eradication in the U.S.A. One of the key factors in such efforts will be the detection of BVDV infections, particularly targeting persistently infected animals. To assess current BVDV detection methods in the U.S.A., 26 veterinary diagnostic laboratories in 23 states were surveyed. Survey questions related to the types of tests currently offered, the number of tests performed, the reasons for test requests, the type of samples used, whether sample pooling was performed, and whether follow-up testing or information regarding bovine viral diarrhea (BVD) management was provided after positive tests. There was no clear consensus on an individual BVDV testing method, the pooling of samples or the retesting of positive animals. Ear-notch antigen capture enzyme-linked immunosorbent assay (ACE) was the test most frequently performed based on the absolute number of tests. However, when the data were adjusted to reflect individual laboratory choices, the number of ACE and immunohistochemistry tests performed on ear notches was nearly equal. Only 55% of diagnostic laboratories provided BVD management information to producers or veterinarians who submitted positive samples. There was no significant difference in the number of positive tests in laboratories that received the majority of their samples for screening purposes versus laboratories that received the majority of their samples because BVDV was suspected based on clinical signs in a herd. PMID- 17121093 TI - Hyperandrogenism from an ovarian interstitial-cell tumor in an alpaca. AB - An 8-year-old intact female Huacaya alpaca (Lama pacos) was presented for recent development of male behavior. Serum testosterone concentration was determined to be 969.1 pg/ml by using radioimmunoassay, while the range in 33 healthy female adult intact alpacas was 11.7-62.1 pg/ml. An ovarian mass was suspected, and an exploratory laparotomy was performed. A tan mass was present on the left ovary. Histologically, the mass was composed of closely packed, plump, polygonal cells with central round nuclei with granular chromatin and abundant eosinophilic finely granular to vesiculate cytoplasm. An ovarian benign interstitial (Leydig) cell tumor was diagnosed. PMID- 17121094 TI - Degenerative myelopathy and vitamin A deficiency in a young black-maned lion (Panthera leo). AB - Degenerative myelopathy and vitamin A deficiency were diagnosed in a 1-year-old, female, black-maned lion (Panthera leo). Diffuse white matter degeneration characterized by dilated myelin sheaths, Wallerian degeneration, and reactive astrocytosis was present at all levels of the spinal cord. With luxol fast blue resyl echt violet stain, bilaterally symmetrical demyelination was observed in the fasciculus cuneatus of the cervical spinal cord and in peripheral white matter of cervical, thoracic, and lumbar segments. Additionally, the ventral gray columns and brain stem nuclei contained rare chromatolytic neurons with abnormal neurofilament accumulation. Leptomeninges of the cervical spinal cord were focally adhered to the dura and thickened by fibrosis and osseous metaplasia. Vitamin A deficiency was diagnosed based on hepatic vitamin A concentration of 1.71 microg/g dry weight. Adequate hepatic vitamin A concentration for yearling to adult domestic animals ranges between 150 and 1000 microg/g dry weight. Lesions were distinct from those previously described in young captive lions with vitamin A deficiency, which had thickened skull bones and cerebellar herniation. The pathogenesis of vitamin A-associated myelopathy in this lion may be similar to that described in adult cattle, which is believed to result from spinal cord compression secondary to elevated pressure of cerebrospinal fluid. PMID- 17121095 TI - Halicephalobus gingivalis-associated meningoencephalitis in a Thoroughbred foal. AB - A 13-week-old Thoroughbred colt from central Kentucky was euthanized after an acute onset of ataxia, blindness, head tremors, leaning to the right, recumbency, and seizures. Microscopically, there was a verminous meningoencephalitis characterized by an eosinophilic and granulomatous inflammatory reaction primarily affecting the cerebellum. Dispersed within regions of inflammation were numerous cross and longitudinal sections of intact and degenerative small nematodes. The nematodes had dorsoflexed ovaries and ventroflexed vulvas, which are distinguishing features of Halicephalobus gingivalis. Intact nematodes, compatible with H. gingivalis, also were recovered and identified from portions of the brain that had been frozen for 5-week post-necropsy examination via tissue maceration and additional laboratory techniques. PMID- 17121096 TI - Systemic blastomycosis in a horse. AB - Progressive multisystemic disease caused by Blastomyces dermatitidis was diagnosed in a 17-year-old Quarter horse broodmare. The mare had been treated unsuccessfully with antibiotics for mastitis 3 months postpartum. The disease progressed to exudative cutaneous lesions affecting the ventrum, pectoral region, and limbs accompanied by weight loss across several months. Yeast bodies were observed in swabs of the cutaneous exudate, suggesting a clinical diagnosis of blastomycosis. Following referral, pleural effusion, cavitated lung lesions, and hyperproteinemia were identified, and the mare was euthanized because of poor prognosis. Necropsy revealed extensive pyogranulomas in the mammary gland, skin, subcutaneous tissues, and lungs, accompanied by thrombi in major blood vessels of the lungs and hind limbs. Histologically, pyogranulomatous inflammation was evident in many tissues, and fungal organisms were seen in sections of mammary gland, skin, subcutis, pericardium, and lung. Blastomyces dermatitidis was cultured from mammary tissue, lungs, lymph node, and an inguinal abscess. Although blastomycosis is endemic in the area of origin of the mare in northwestern Wisconsin, the disease is extremely rare in horses and hence easily misdiagnosed. Unique features of this case included the extent of mammary gland involvement and the presence of thrombi in multiple sites. PMID- 17121097 TI - Meningoencephalitis in an adult cow due to Mortierella woifli. AB - A 7-year-old dairy cow presented with clinical signs of neurologic disease. Despite treatment with penicillin, the cow died 36 hours after initial presentation. Necropsy examination revealed multiple foci of hemorrhage within the cerebrum and thickened meninges. Additionally, endometritis and consolidation of approximately 30% of both lungs was observed. Histology revealed necrotizing vasculitis, infarction, and hemorrhage within sections of the brain, uterus, and lung. Large numbers of intralesional fungal hyphae were visible. Because only formalin-fixed tissue was available, polymerase chain reaction was used to make an etiologic diagnosis of Mortierella wolfii. PMID- 17121098 TI - Morphological changes in the lungs of meconium-stained piglets. AB - Meconium staining of the skin is a common event associated with fetal hypoxia, stillbirths, weak-born piglets, and neonatal mortality. Aspiration of meconium leads to meconium aspiration syndrome (MAS). This study was undertaken to assess the relationship between the degree of meconium staining of the skin at birth, meconium aspiration, and pulmonary changes in porcine neonates. A total of 353 farrowing sows and 3,693 born piglets were monitored during parturition and for 15 days after delivery. Umbilical cords were classified as normal or ruptured. Meconium staining in the skin was graded as nonstained, mildly, moderately, and severely stained. Mortality from birth to 15 days of age was 8.4%. The lungs from 60 meconium-stained piglets and 60 lungs from nonstained piglets were collected and microscopically examined for meconium aspiration and inflammation. Rupture of the umbilical cord was significantly higher (P < 0.01) in meconium-stained piglets. Microscopically, 32% and 40% of the lungs had evidence of meconium for the stained and nonstained groups, respectively. The microscopic grade of meconium aspiration and inflammatory cells was not different between nonstained and meconium-stained piglets. Aspiration of meconium induced a granulomatous response in the lungs. It was concluded that the grade of meconium staining is a good indicator of fetal hypoxia, but not a good predictor for meconium aspiration and MAS in piglets. PMID- 17121099 TI - Spontaneous subcutaneous myxosarcoma in a captive European hedgehog (Erinsceus europaeus). AB - A surgically excised biopsy representing a subcutaneous mass on the left side of the neck from a 3-year-old female European hedgehog (Erinsceus europaeus) was presented. Spontaneous myxosarcoma was diagnosed based on histological, immunohistochemical, and ultrastructural characteristics. The neoplasm grossly consisted of a firm, pale, multilobulated mass with a characteristic clear gelatinous fluid. Histologically, the neoplasm was nonencapsulated and composed of pleomorphic stellate or spindle-shaped vimentin and periodic acid-Schiff positive cells arranged in loose sheets and occasionally whorls. The neoplastic cells were suspended in Alcian blue-positive stroma and contained infrequent mitotic figures. Evidence of a viral etiology was not detected using electron microscopy and polymerase chain reaction. This is the first case report of a myxosarcoma in a captive European hedgehog. PMID- 17121100 TI - From conflict to consensus: managing competing interests in your organization. PMID- 17121101 TI - Executive decision making. Effective processes for making and implementing decisions. PMID- 17121102 TI - The prevalence and impact of disability among healthcare executives. PMID- 17121103 TI - Ethical implications of transparency. Valid justification is required when withholding information. PMID- 17121104 TI - Lean processes improve patient care. Observing processes through patients' eyes can lead to improvements. PMID- 17121105 TI - Meeting the language challenge. One organization's quest for better understanding. PMID- 17121106 TI - Payment rule changes. It pays to understand changes affecting other providers. PMID- 17121107 TI - Going Retail. New leadership skills to keep up with healthcare's market shift. PMID- 17121108 TI - Career and competency mapping. Leaders embrace life-long learning for success. PMID- 17121109 TI - Putting the 'I' in service. Management processes help staff connect work to patient outcomes. PMID- 17121110 TI - Embracing innovation. Solving key challenges provides future solutions. PMID- 17121112 TI - Zero waste--a desirable goal in residue utilization? PMID- 17121111 TI - Prepare the next generation of healthcare leaders. Make a connection with health management students. PMID- 17121113 TI - PCDD/F and related compounds in solid residues from municipal solid waste incineration--a literature review. AB - Emissions of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F) from waste incineration into the air have been a major focus of interest during the last two decades. An integrated approach to clean waste disposal has to take the occurrence of PCDD/F in all residues into account. This paper compiles published data on concentration ranges of PCDD/F and the related compounds polychlorinated biphenyls (PCB), chlorinated benzenes and phenols, as well as polyaromatic hydrocarbons (PAH) in solid residues from waste incineration in grate furnaces and their development since 1985. A short description of inertization processes for PCDD/F loaded materials is added. PMID- 17121114 TI - Predicted growth of world urban food waste and methane production. AB - Landfill gas emissions are one of the largest anthropogenic sources of methane especially because of food waste (FW). To prevent these emissions growing with world population, future FW best management practices need to be evaluated. The objective of this paper was therefore to predict FW production for 2025 if present management practices are maintained, and then, to compare the impact of scenario 1: encouraging people to stay in rural areas and composting 75% of their FW, and; of scenario 2, where in addition to scenario 1, composting or anaerobically digesting 75% of urban FW (UFW). A relationship was established between per capita gross domestic product (GDP) and the population percentage living in urban areas (%UP), as well as production of municipal solid waste (MSW) and UFW. With estimated GDP and population growth per country, %UP and production of MSW and UFW could be predicted for 2025. A relatively accurate (R(2) > 0.85) correlation was found between GDP and %UP, and between GDP and mass of MSW and FW produced. On a global scale, MSW and UFW productions were predicted to increase by 51 and 44%, respectively, from 2005 to 2025. During the same period, and because of its expected economic development, Asia was predicted to experience the largest increase in UFW production, of 278 to 416 Gkg. If present MSW management trends are maintained, landfilled UFW was predicted to increase world CH4 emissions from 34 to 48 Gkg and the landfill share of global anthropogenic emissions from 8 to 10%. In comparison with maintaining present FW management practices, scenario 1 can lower UFW production by 30% and maintain the landfill share of the global anthropogenic emissions at 8%. With scenario 2, the landfill share of global anthropogenic emissions could be further reduced from 8 to 6% and leachate production could be reduced by 40%. PMID- 17121115 TI - Generation amount prediction and material flow analysis of electronic waste: a case study in Beijing, China. AB - The draft legislation on e-waste prepared by the Chinese national government assigns management responsibility to local governments. It is an urgent task for the municipal government to plan an effective system as soon as possible to divert the e-waste flow from the existing informal e-waste recycling processes. This paper presents a case study implemented in Beijing, the capital city of China, with the purpose of predicting the amount of obsolete equipment for five main kinds of electronic appliances from urban households and to analyse the flow after the end of their useful phase. The amount to be handled was 885,354 units in 2005 and is predicted to double by 2010. Due to consumption growth and the expansion of urbanization it is estimated that the amount will increase to approximate 2,820,000 units by 2020: 70% of the obsolete appliances will be awaiting collection for possible recycling, 7% will be stored at the owner's home for 1 year on average and 4% will be discarded directly and enter the municipal solid waste collecting system. The remaining items will be reused for about 3 years on average after the change of ownership. The results of this study will assist the waste management authorities of Beijing to plan the collecting system and facilities needed for management of e-waste generated in the near future. PMID- 17121116 TI - Effects of calcium hydroxide and calcium chloride addition to bentonite in iron ore pelletization. AB - Pyrite ash is created as waste from the roasting of pyrite ores during the production of sulphuric acid. These processes generate great amounts of pyrite ash waste that is generally land filled. This creates serious environmental pollution due to the release of acids and toxic substances. Pyrite ash waste can be utilized in the iron production industry as a blast furnace feed to process this waste and prevent environmental pollution. The essential parameters affecting the pelletization process of pyrite ash were studied using bentonite as a binder. Experiments were then carried out using bentonite and a mixture of bentonite with calcium hydroxide and calcium chloride in order to make the bentonite more effective. The metallurgical properties of pyrite ash, bentonite, calcium hydroxide, calcium chloride, a mixture of these and sintered pellets were studied using X-ray analysis. The crushing strength tests were carried out to investigate the strength of pyrite ash waste pellets. The results of these analyses showed that pyrite ash can be agglomerated to pellets and used in the iron production industry as a blast furnace feed. The crushing strength of the pellets containing calcium hydroxide and calcium chloride in addition to bentonite was better than the strength of pellets prepared using only bentonite binder. PMID- 17121117 TI - Feasibility study of a passive aeration reactor equipped with vertical pipes for compost stabilization of cow manure. AB - Pilot-scale composting was carried out with cow manure to evaluate the performances of two passive aeration systems: a conventional passive aeration system equipped with horizontal pipes and an unusual passive aeration method based on air delivery by means of vertical pipes. The effects of both types of passive aeration apparatus were investigated in order to determine the degree of composting rate by continuously monitoring temperature, moisture content, organic matter, electrical conductivity, pH and C/N ratio in the piles. Temperatures in the range of thermophily (55-65 degrees C) were reached in all runs within 1-2 days then lasting for about 1 week, a span long enough for pathogen abatement. Results suggest that passive aeration carried out by vertical pipes is more effective for air delivery into compost piles than conventional passive aeration of air adduction with horizontal pipes. The variation in the number of vertical pipes was revealed to be an important parameter for the control of composting rate and temperature. Composting rates estimated from the heat balance equation were substantially in agreement with those computed through the conversion ratio of total organic matter decrement. The conversion ratios and composting rates obtained in this study using passive aeration with vertical pipes were well aligned with those found using forced air delivery systems. PMID- 17121118 TI - Household, hotel and market waste audits for composting in Vietnam and Laos. AB - In Da Nang and Ha Long, Vietnam and in Vientiane, Laos, there was interest by local authorities in separating and composting waste in order to reduce environmental and health problems at the local landfills and to produce a soil conditioner for local agricultural use. To assist in the planning of composting projects, three studies were carried out to estimate waste quantities and composition. 1. A 9-day audit of waste from 45 vendors in a market in Vientiane, the capital of Laos. The total quantity of waste and the quantity in each of nine categories were estimated for each of six different types of vendors. 2. A 7-day audit of waste disposed by three hotels in the tourist area of Ha Long, Vietnam. Waste quantities were estimated in total, on a per guest basis, and in three main categories: compostables, recyclables and miscellaneous. 3. A 7-day audit of waste collected from 74 households in Da Nang, the fourth largest city in Vietnam. Waste from each household was separated into compostable and non compostable waste. Over 60% of each waste source comprised compostable waste and this was considered significant enough to warrant further planning of composting operations. PMID- 17121119 TI - Evaluating effects of wind-induced pressure fluctuations on soil-atmosphere gas exchange at a landfill using stochastic modelling. AB - The impact of wind turbulence-induced pressure fluctuations at the soil surface on landfill gas transport and emissions to the atmosphere at an old Danish landfill site was investigated using stochastic modelling combined with soil property and gas transport data measured at the site. The impacts of soil physical properties (including air permeability and volumetric water content) and wind-induced pressure fluctuation properties (amplitude and temporal correlation) on landfill gas emissions to the atmosphere were evaluated. Soil-air permeability and pressure fluctuation amplitude were found to be the most important parameters. Wind-induced gas emissions were further compared with gas emissions caused by diffusion and by long-term pressure variations (due to passing weather systems). Here diffusion and wind-induced gas transport were found to be equally important with wind-induced gas transport becoming the most important at lower soil-air contents. PMID- 17121120 TI - Graded landfill requirements in South Africa--the climatic water balance classification. AB - Landfilling in South Africa is controlled by a set of statutory Minimum Requirements, based on a landfill classification system. Landfills are classified according to the type of waste, the projected final size of the landfill and the climate. Climate is important as climatic conditions in South Africa vary from humid sub-tropical in the east, to semi-arid on the central plateau, to semi desert in the west. Anti-pollution measures are closely related to climate and size, with the pollution potential of small, general (i.e. domestic) waste landfills in dry climates being regarded as negligible. At the other end of the scale, large landfills where hazardous wastes are disposed have a serious pollution potential and must be designed as containment systems. The paper describes the method currently in use for deciding on the climatic classification of a site, followed by the new method that will be adopted when the latest revision of the Minimum Requirements appears shortly. PMID- 17121121 TI - Assessment of long-term pH developments in leachate from waste incineration residues. AB - Environmental assessment of residue disposal needs to account for long-term changes in leaching conditions. Leaching of heavy metals from incineration residues are highly affected by the leachate pH; the overall environmental consequences of disposing of these residues are therefore greatly influenced by changes in pH over time. The paper presents an approach for assessing pH changes in leachate from municipal solid waste incineration (MSWI) air-pollution-control (APC) residues. Residue samples were subjected to a stepwise batch extraction method in order to obtain residue samples at a range of pH values (similar to common pH-dependence tests), and then on these samples to determine leaching of alkalinity as well as remaining solid phase alkalinity. On a range of APC residues covering various pretreatment and disposal options, this procedure was used to determine leachable and residual alkalinity as a function of pH. Mass balance calculations for typical disposal scenarios were used to provide data on pH as a function of the liquid-to-solid (L/S) ratio in the leaching system. Regardless of residue type and pretreatment, pH was found to stay above 7 for L/S ratios up to about 2000 L kg(-1) corresponding to about 100,000 years in typical landfill scenarios. It was found that pH changes were mainly governed by alkalinity decreases from leaching processes rather than neutralization reactions. The results suggest that leaching testing for assessment purposes should be carried out in the alkaline range, for example, at pH 9. The paper offers a thorough basis for further modelling of incineration residue leaching and for modelling the environmental consequences of landfilling and utilization of these residues. PMID- 17121122 TI - Linear accelerator radiosurgery for vestibular schwannomas. PMID- 17121123 TI - Linear accelerator radiosurgery for vestibular schwannomas. AB - OBJECT: Radiosurgery has become a popular treatment for small vestibular schwannomas (VSs). The aim of this study was to review an extensive, single institution experience with linear accelerator (LINAC) radiosurgery for VSs. METHODS: Between July 1988 and August 2005, 390 patients with VSs were treated with LINAC-based radiosurgery at the authors' institution. Patient and treatment variables were prospectively maintained in a computer database. Outcomes were tracked through periodic clinical examinations and annual scanning studies. Multivariate and actuarial statistics were used to analyze rates of local tumor control and complications, including facial and trigeminal neuropathies, after treatment. One- and 2-year actuarial control rates were both 98%, and the 5-year actuarial control rate was 90%. Only four patients (1%) required surgery for tumor growth. Seventeen patients (4.4%) reported facial weakness and 14 patients (3.6%) reported facial numbness after radiosurgery. The risk of these complications rose with increasing tumor volume or increasing radiosurgical dose to the tumor periphery. Since 1994, when doses were deliberately lowered to 1250 cGy, only two patients (0.7%) have experienced facial weakness and two (0.7%) have experienced facial numbness. CONCLUSIONS: Radiosurgery provides a safe and effective therapeutic alternative to surgery for small VSs. PMID- 17121124 TI - Endovascular coil embolization in patients with poor neurological grades after aneurysmal rupture. PMID- 17121125 TI - Contribution of endovascular therapy to the management of poor-grade aneurysmal subarachnoid hemorrhage: Clinical and angiographic outcomes. AB - OBJECT: Treatment of patients presenting with poor-grade (Hunt and Hess Grade IV or V) subarachnoid hemorrhage (SAH) is controversial. Endovascular coil embolization has been considered a valuable therapeutic alternative to surgical clip placement for this kind of patient. The aim of the present study was to evaluate immediate and long-term angiographic and clinical outcomes in patients with poor-grade SAH treated by endovascular embolization. METHODS: One hundred eleven patients with Hunt and Hess Grade IV or V SAH were treated with endovascular embolization at the University of California at Los Angeles Medical Center between October 1990 and December 2004. Eighty patients harbored Grade IV hemorrhages and 31 patients had Grade V ones. Immediate and long-term anatomical and clinical outcomes were evaluated in all patients. Long-term clinical outcome assessments were based on follow-up data obtained over an average of 32 months posttherapy. Technical complications occurred in 15 patients (13.5%). Immediate complete aneurysm occlusion was observed in 51.4% of aneurysms. Angiographic, long-term follow-up review revealed aneurysm recanalization in 16.2% of cases. Thirty-nine patients (35.1%) demonstrated a favorable long-term clinical outcome. The overall mortality rate in this patient series was 32.4%. The mortality rate associated with vasospasm was significantly higher in patients with Grade IV SAHs than in those with Grade V hemorrhages. CONCLUSIONS: The results of this study demonstrate a valuable contribution of endovascular therapy of ruptured intracranial aneurysms in patients with Hunt and Hess Grade IV or V SAH. This technique was successful in decreasing repeated aneurysm rupture and in enabling aggressive medical management during the acute phase of SAH. This is particularly important in patients with Grade IV SAH because of their potential for obtaining higher physical and functional recoveries. PMID- 17121126 TI - Risk of cerebral vasopasm after subarachnoid hemorrhage reduced by statin therapy: A multivariate analysis of an institutional experience. AB - OBJECT: Impairment of endothelial nitric oxide synthase (eNOS), endothelium dependent relaxation, and cerebrovascular autoregulation all occur in vasospastic cerebral arteries following subarachnoid hemorrhage (SAH). The 3-hydroxy-3 methylglutaryl coenzyme A reductase inhibitors, or statins, both improve endothelial function and increase eNOS messenger RNA, protein, and enzymatic activity threefold. Increasing experimental evidence in animal models of SAH suggests that statins may ameliorate cerebral vasospasm. The authors hypothesized that patients chronically treated with statins would have a decreased risk of symptomatic vasospasm after SAH. METHODS: The authors retrospectively reviewed the charts of 115 patients with SAH who were consecutively admitted to the Neuroscience Intensive Care Unit of Duke University between 1998 and 2001. The independent association of statin therapy to symptomatic vasospasm was assessed using multivariate logistic regression analysis. Fifteen patients (13%) admitted with SAH were receiving statin therapy for at least 1 month before admission. Forty-nine patients (43%) experienced symptomatic vasospasm a mean of 5.8 +/- 3 days after onset of SAH. Current statin therapy on admission (odds ratio [OR] 0.09, 95% confidence interval [CI] 0.01-0.77) was independently associated with an 11-fold reduction in the risk of symptomatic vasospasm. Fisher Grade 3 SAH (OR 2.82, 95% CI 1.50-5.71) and rupture of anterior cerebral or internal carotid artery aneurysm (OR 3.77, 95% CI 1.29-10.91) were independently associated with an increased risk of symptomatic vasospasm. CONCLUSIONS: In this retrospective case series, patients who received statin therapy for at least 1 month demonstrated an 11-fold decrease in the risk of developing symptomatic vasospasm after SAH. PMID- 17121127 TI - Monitoring of muscle motor evoked potentials during cerebral aneurysm surgery: intraoperative changes and postoperative outcome. AB - OBJECT: The authors in this study evaluated muscle motor evoked potentials (MMEPs) elicited by transcranial electrical stimulation (TES) and direct cortical stimulation as a means of monitoring during cerebral aneurysm surgery. The analysis focused on the value and frequencies of any intraoperative changes and their correlation to the postoperative motor status. METHODS: One hundred nineteen patients undergoing surgery for 148 cerebral aneurysms were included in the study. Muscle motor evoked potentials were elicited by a train of five constant-current anodal stimuli with an individual pulse duration of 0.5 msec and a stimulation rate of 2 Hz. Stimulation intensity was up to 240 mA for TES and up to 33 mA for direct cortical stimulation. The MMEPs were continuously recorded from the abductor pollicis brevis and tibialis anterior muscles bilaterally and from the biceps brachii and extensor digitorum communis muscles contralateral to the surgical side. The motor status was evaluated immediately after surgery and 7 days later. In 97% of the patients MMEPs were recordable for continuous neurophysiological monitoring of the vascular territory of interest throughout the surgery. In 14 patients significant intraoperative MMEP changes occurred, resulting in a transient motor deficit in one patient and a permanent motor deficit in six. The permanent loss of MMEPs in three patients was followed by a permanent severe motor deficit in one patient and severe clinical deterioration in the other two. CONCLUSIONS: Data in this study demonstrated that MMEPs are a useful means of intraoperative neurophysiological monitoring of motor pathway integrity and predicting postoperative motor status. The intraoperative loss of MMEPs reliably predicts both severe and permanent postoperative motor deficits. PMID- 17121128 TI - Multislice computed tomography angiography screening for new aneurysms in patients with previously clip-treated intracranial aneurysms: Feasibility, positive predictive value, and interobserver agreement. AB - OBJECT: Multislice computed tomography (CT) angiography may be useful for screening patients with intracranial aneurysms that are treated with clip occlusion. However, cobalt clips produce much more artifact on CT scans than titanium clips, which may hamper the evaluation of the image obtained at the clip site. METHODS: The authors screened 415 patients with previously ruptured aneurysms that had been treated using cobalt clips. Screening was performed using multislice CT angiography. The feasibility of this modality for screening these patients (based on the complication risk, CT angiography quality, and artifact avoidance) and interobserver agreement were evaluated. Patients in whom the presence of an aneurysm was suspected based on results of CT angiography studies underwent digital subtraction (DS) angiography. False-negative and false-positive findings were recorded, and the positive predictive value (PPV) was calculated. Eight patients (1.9%) had allergies to the contrast material. The quality of the CT angiography image was suboptimal in 14%. In 52%, clip artifacts hampered evaluation of the clip site. In 65 patients who underwent DS angiography, there were nine false-positive and eight false-negative reports related to aneurysms that were either small, located at the clip site, or were infundibula. The PPV on a per-patient basis was 86% (95% confidence interval [CI] 75-94%); for aneurysms at the clip site it was 83% (95% CI 61-95%); and for aneurysms at different locations it was 91% (95% CI 81-97%). The interobserver agreement was good (kappa = 0.69; 95% CI 0.60-0.78). CONCLUSIONS: Except for the evaluation of images from the clip site, CT angiography has good feasibility with good PPV and interobserver agreement. Drawbacks are that very small aneurysms can be missed and that visualization is poor at the clip site in patients in whom cobalt clips have been placed for occlusion. This second problem can be expected to resolve with the increasing use of titanium clips. PMID- 17121129 TI - Radiosurgical treatment for rolandic arteriovenous malformations. AB - OBJECT: The authors reviewed the radiosurgical outcomes in patients with arteriovenous malformations (AVMs) located in the rolandic area, including the primary motor and sensory gyri. METHODS: The study population consisted of 38 patients with rolandic-area AVMs who underwent linear accelerator radiosurgery at the University of Toronto between 1989 and 2000. Obliteration rate, risk of hemorrhage during the latency period, radiation-induced complications, seizure control, and functional status were evaluated. Patients were also divided into two subgroups according to AVM volume (< 3 cm3 and > or = 3 cm3). Patients were followed up for a median of 42.4 months (range 30-103 months), and the median age of the patients was 40 years (range 12-67 years). The median AVM volume was 8.1 cm3 (range 0.32-21, mean 8.32 cm3), and the median dose at the tumor margin was 15 Gy (range 15-22, mean 16.8 Gy). The risk of hemorrhage after radiosurgery was 5.3% for the 1st year, 2.6% for the 2nd, and 0% for the 3rd. Two patients (5.3%) sustained adverse effects related to radiation for more than 6 months. Complete nidus obliteration after a single radiosurgical treatment was achieved in 23 patients (60.5%). The obliteration rate for AVMs smaller than 3 cm3 was 83.3% (10 of 12) and that for AVMs larger than or equal to 3 cm3 was 50% (13 of 26). Among the patients who had seizures as the initial presentation, 51.8% were free of seizures after radiosurgery and the seizure pattern improved in 40.7% during the 3rd and last year of follow up. Overall, excellent results (obliteration and no new or worsening neurological deficit) can be achieved in approximately 60% of patients. This percentage varies according to the AVM size and can reach 83% in patients with AVMs smaller than 3 cm3. CONCLUSIONS: Radiosurgery is a safe and effective treatment for people with rolandic AVMs. The low rate of morbidity associated with radiosurgery, compared with other treatments, indicates that this method may be the first choice for patients with AVMs located in this area. PMID- 17121130 TI - Split facial nerve course in vestibular schwannomas. AB - OBJECT: The facial nerve in vestibular schwannomas (VSs) is located on the ventral tumor surface in more than 90% of cases; other courses are rare. A split facial nerve course with two distinct bundles has thus far been described exclusively for medial extrameatal tumors. METHODS: Between 1996 and 2005, 16 consecutive cases of 241 surgically treated VSs were observed to have distinct splitting of the facial nerve. The mean tumor size measured 27 mm. In one third of the cases, intrameatal tumor extension with obliteration of the fundus was documented. All patients underwent extensive intraoperative neurophysiological monitoring using multichannel electromyography recordings. Patients were reevaluated 12 months after surgery. In all 16 patients, distinct splitting of the facial nerve was demonstrated. The major portion of the facial nerve followed a typical course on the ventral tumor surface. The smaller nerve portion in all cases ran parallel to the brainstem up to the level of the trigeminal root exit zone and crossed on the cranial tumor pole to the internal auditory canal. The two nerve portions rejoined at the level of the porus acusticus. The smaller portion carried fibers exclusively to the orbicularis oris muscle, whereas the major portion supplied all three branches of the facial nerve. CONCLUSIONS: In VSs, an aberrant course with distinct splitting of the facial nerve adds considerably to the surgical challenge. Long-term facial nerve results are excellent with extensive neurophysiological monitoring, which allows the differentiation and identification of aberrant facial nerve fibers and avoids additional risks to facial nerve preservation. PMID- 17121131 TI - Intrasaccular combination of metallic coils and onyx liquid embolic agent for the endovascular treatment of cerebral aneurysms. AB - OBJECT: The aim of this study was to report on a novel technique in which metallic embolization coils were combined with the Onyx liquid embolic agent in the aneurysm sac to achieve a more durable result after endovascular treatment. This therapeutic procedure was performed in selected cases in which, based on the authors' experiences, either coil embolization or Onyx alone would likely have failed. The authors report long-term clinical and angiographic follow-up results in 20 consecutive intracranial aneurysms treated using this combination for defined indications. METHODS: Twenty aneurysms in 20 patients were treated with a combination of embolic coils and Onyx. Four aneurysms were giant; 13, large; and three, small. This new technique was used when standard Onyx or coil treatment with balloon assistance was determined to involve a higher possibility of recanalization, because either an adjunctive stent insertion could not be performed or the Onyx technique could not be used due to an unsuccessful seal test or intraaneurysm balloon prolapse. In one case, an adjunctive stent was placed before coil placement and Onyx deposition to control the material in the sac of the aneurysm, which had a fusiform neck. All aneurysms were completely occluded after using this technique. No clinical or technical adverse events occurred in any of the cases. Follow-up angiography was performed in all patients: 3-year studies in six patients, 2-year studies in five, and 1-year studies in nine. None of these studies demonstrated aneurysm regrowth or parent artery occlusion. CONCLUSIONS: The combination of the embolic coils and the Onyx liquid embolic agent provides very durable aneurysm occlusion for defined indications. PMID- 17121132 TI - Reliability of cobalt-chromium alloy aneurysm clips after long-term implantations in patients with cerebral aneurysms. AB - OBJECT: Aneurysm clip reliability after long-term implantation in vivo has not been examined. In this study the authors evaluated the mechanical properties and surface elemental composition of Co-Cr alloy aneurysm clips implanted for more than 10 years in patients with cerebral aneurysms. METHODS: Five aneurysm clips implanted for ruptured or unruptured intracranial aneurysms were retrieved and examined. New aneurysm clips were applied to the regrown aneurysms. The implantation period ranged from 11 to 20 years. Four new and unused aneurysm clips were also examined as controls. The mechanical properties of the clips were tested by measuring their closing force and bending strength. The surface elemental composition of the aneurysm clips was evaluated using x-ray photoelectron spectroscopy. The closing force of the retrieved clips exceeded the minimum force requirement at the time of manufacture. The bending strength was similar between the retrieved and control clips. Chromium oxide was the predominant constituent on the surface of all clips, and its concentration on the retrieved clips was higher than that on the control clips. CONCLUSIONS: Data in the present study demonstrated that Co-Cr alloy aneurysm clips retain their mechanical properties in vivo for a long time, which indicates the reliability of these clips. PMID- 17121133 TI - Using endothelial nitric oxide synthase gene polymorphisms to identify intracranial aneurysms more prone to rupture in Japanese patients. AB - OBJECT: Recent investigators found that the presence of three tandem polymorphisms of the endothelial nitric oxide synthase (eNOS) gene-promoter single nucleotide polymorphism (SNP) T-786C, intron-4 27-bp variable number of tandem repeats, and the G894T SNP in exon 7-was indicative of intracranial aneurysms more prone to rupture in a Caucasian patient sample. In the present study, the authors sought to determine whether the presence of these eNOS polymorphisms could indicate which Japanese patients with aneurysms were more endangered by a subarachnoid hemorrhage (SAH). METHODS: The three eNOS polymorphisms were genotyped in 297 patients with ruptured aneurysms (RAs), 108 patients with unruptured aneurysms (UAs), and 176 healthy volunteers by using polymerase chain reaction. The distribution of the variant alleles did not differ significantly (p > 0.05) between the RA group and the UA group. The frequency of the corresponding genotypes between the two groups and a haplotype analysis did not show any significant differences. Further comparisons of the RA and UA groups with the control group did not yield any significant allele or genotype frequency differences. Conclusions. These data show that the examined set of eNOS polymorphisms were not indicative of which Japanese patients with intracranial aneurysms would suffer an SAH. The presence of eNOS polymorphisms is not useful in identifying intracranial aneurysms that are more prone to rupture in a Japanese patient sample. PMID- 17121134 TI - Magnesium infusion for vasospasm prophylaxis after subarachnoid hemorrhage. AB - OBJECT: Despite the application of current standard therapies, vasospasm continues to result in death or major disability in patients treated for ruptured aneurysms. The authors investigated the effectiveness of continous MgSO4 infusion for vasospasm prophylaxis. METHODS: Seventy-six adults (mean age 54.6 years; 71% women; 92% Caucasian) were included in this comparative matched-cohort study of patients with aneurysmal subarachnoid hemorrhage on the basis of computed tomography (CT) findings. Thirty-eight patients who received continuous MgSO4 infusion were matched for age, race, sex, treatment option, Fisher grade, and Hunt and Hess grade to 38 historical control individuals who did not receive MgSO4infusion. Twelve grams of MgSO4 in 500 ml normal saline was given intravenously daily for 12 days if the patient presented within 48 hours of aneurysm rupture. Vasospasm was diagnosed on the basis of digital substraction angiography, CT angiography, and transcranial Doppler ultrasonography, and evidence of neurological deterioration. Symptomatic vasospasm was present at a significantly lower frequency in patients who received MgSO4 infusion (18%) compared with patients who did not receive MgSO4 (42%) (p = 0.025). There was no significant difference in mortality rate at discharge (p = 0.328). A trend toward improved outcome as measured by the modifed Rankin Scale (p = 0.084), but not the Glasgow Outcome Scale (p = 1.0), was seen in the MgSO4 treated group. CONCLUSIONS: Analysis of the results suggests that MgSO4 infusion may have a role in cerebral vasospasm prophylaxis if therapy is initiated within 48 hours of aneurysm rupture. PMID- 17121135 TI - Does dose rate affect efficacy? The outcomes of 256 gamma knife surgery procedures for trigeminal neuralgia and other types of facial pain as they relate to the half-life of cobalt. AB - OBJECT: Gamma Knife surgery (GKS) is a treatment option for patients with refractory typical trigeminal neuralgia (TN), TN with atypical features, and atypical types of facial pain. The Gamma Knife's 201 60Co sources decay with a half-life of 5.26 years. The authors examined whether the decrease in dose rate over 4.6 years between Co source replacements affected the control rates of facial pain in patients undergoing GKS. METHODS: The authors collected complete follow-up data on 239 of 326 GKS procedures performed in patients with facial pain. Patients were classified by their type of pain. The isocenter of a 4-mm collimator helmet was targeted at the proximal trigeminal nerve root, and the dose (80-90 Gy) was prescribed at the 100% isodose line. Patients reported the amount of pain control following radiosurgery by answering a standardized questionnaire. Eighty percent of patients experienced greater than 50% pain relief, and 56% of patients experienced complete pain relief after GKS. Neither dose rate nor treatment time was significantly associated with either the control rate or degree of pain relief. A significant association between the type of facial pain and the pain control rate after GKS was observed (p < 0.001; Pearson chi-square test). In their statistical analysis, the authors accounted for changes in prescription dose over time to prevent the dose rate from being a confounding variable. There was no observable effect of the dose rate or of the treatment duration within the typical period to source replacement. CONCLUSIONS: Patients with facial pain appear to receive consistent treatment with GKS at any time during the first half-life of the Co sources. PMID- 17121136 TI - Oncocytic meningiomas: Cases with benign histopathological features and a favorable clinical course. AB - OBJECT: Oncocytic meningioma has recently been recognized as a distinct morphological variant of intracranial meningothelial neoplasms, and only a few cases have been reported in the literature. The first description of this lesion, which was based on data in six cases, revealed a potentially aggressive nature with a tendency to infiltrate the brain and to recur. However, the true behavior of, and the long-term follow-up data for, such lesions must still be outlined. METHODS: The authors report on five cases of intracranial oncocytic meningiomas. On neuroimaging, the lesions showed the characteristic features of common meningiomas. All patients underwent gross-total removal of the mass together with the adjacent dura mater. No additional treatments were administered. Histologically, the tumors were composed of sheets, nests, and cords of large polygonal neoplastic cells with finely granular cytoplasm. Necrosis was absent in all cases. Mitosis was also absent or exceedingly rare, and no brain cortex infiltration was observed. The follow up ranged from 6 to 54 months (mean 32.4 months). At the last follow-up evaluation, all patients were asymptomatic and magnetic resonance imaging examinations demonstrated no evidence of tumor recurrence. CONCLUSIONS: Data in the presented cases did not confirm a previously described propensity to aggressiveness in this meningioma subtype. In fact, the histological features as well as the long-term favorable clinical course may suggest benign behavior of such neoplasms, as in the common forms of meningiomas. PMID- 17121137 TI - Genetic profiling of a distant second glioblastoma multiforme after radiotherapy: Recurrence or second primary tumor? AB - OBJECT: In nearly all patients with glioblastoma multiforme (GBM) a local recurrence develops within a short period of time. In this paper the authors describe two patients in whom a second GBM developed after a relatively long time interval at a site remote from the primary tumor. The genetic profiles of the tumors were compared to discriminate between distant recurrence and a second primary tumor. METHODS: Both patients harboring a supratentorial GBM were treated with surgery and local high-dose radiotherapy. Local control of the disease at the primary tumor site was achieved. Within 2 years, a second GBM developed in both patients, not only outside the previously irradiated target areas but infratentorially in one patient and in the opposite hemisphere in the other. The tumors were examined for the presence of several genetic alterations that are frequently found in GBMs--a loss of heterozygosity at chromosome regions 1p36, 10pl5, 19q13, and 22q13, and at the CDKN2A, PTEN, DMBT1, and TP53 gene regions; a TP53 mutation; and EGFR amplification. In the first patient, genetic profiling revealed that the primary tumor had an allelic imbalance for markers in several chromosome regions for which the second tumor displayed a complete loss. In the second patient, genetic profiling demonstrated the presence of genetic changes in the second tumor that were identical with and additional to those found in the primary tumor. CONCLUSIONS: Based on the similarities between the genetic profiles of the primary and the second tumors in these patients, the authors decided that in each case the second distant GBM was a distant recurrence rather than a second independent primary tumor. PMID- 17121138 TI - Evaluation of particulate systems supporting tumor cell fractions in a preventive vaccination against intracranial rat glioma. AB - OBJECT: Irradiated autologous tumor cells are commonly used as a source of antigens in antiglioma vaccinations to activate the immune system. As cell number is often a limiting factor in these cells' preparation, the aim of the present study was to find a means that can lower the amount of cells required. Among strategies currently developed, adjuvant particulate systems offer a promising means to improve the antitumor immune response. In this study, the authors were interested in evaluating the role of particulate systems containing biodegradable microspheres that carry tumor cell fractions on their surfaces in the induction of a protective immunity in the 9L/Fischer 344 rat glioma model. The efficiency of these particulate systems was compared to that of irradiated 9L cells. METHODS: Particulate systems composed of poly(D,L-lactide-co-glycolide) (PLGA) microspheres that support 9L cell fractions on their surfaces (cell lysates or plasma membranes) or irradiated 9L cells alone were injected subcutaneously into the flanks of syngeneic Fischer 344 rats. Eighteen days later, the rats were intracranially injected with nonirradiated 9L cells. A study of survival in these animals and an analysis of the resulting immune response were then conducted. For the same amount of protein (50 microg) injected, irradiated 9L cells provided long-term survival in 30% of animals, whereas 9L plasma membranes adsorbed onto PLGA microspheres provided long-term survival in 10% of animals and cell lysates adsorbed onto microspheres provided long-term survival in 0%. Accordingly, particulate systems induced a lower T helper cell Type 1 (Th1) peripheral immune response than irradiated 9L cells. However, greater secretion of Th1 cytokines was observed when particulate systems were used than when cell fractions separated from microspheres were used, indicating the adjuvant property of these particulate systems. CONCLUSIONS: Particulate systems have adjuvant properties but are still less efficient than irradiated whole tumor cells for vaccinations. Encapsulation of an activating molecule in the microsphere will be the next developmental step in the search for efficient antiglioma vaccinations. PMID- 17121139 TI - Jugular tubercle: Morphometric analysis and surgical significance. AB - OBJECT: Maximizing intradural exposure via the extreme lateral infrajugular transcondylar-transtubercular exposure (ELITE) approach depends on understanding the fundamental anatomy of the jugular tubercle (JT). Drilling the JT can maximize the extent of exposure achieved with the ELITE approach. Removing the JT is critical for optimizing access to the inferior and midclival areas, vertebrobasilar artery junction, and ventral pons and medulla. METHODS: In this cadaveric study, the individual structural variations in the JT were evaluated in 100 split occipital bones. The mean length of the JT was 1.65 +/- 0.36 cm (range 1.2-3 cm); its mean width was 1.15 +/- 0.16 cm (range 0.7-1.7 cm); and its mean thickness was 0.61 +/- 0.15 cm (range 0.2-1 cm). The authors analyzed the difference in morphometric data with regard to right and left sides and found no statistically significant difference between the two sides. Furthermore, data from the morphometric study were compared with the results of 20 measurements obtained from three-dimensional computed tomography (3D CT) scans. Accordingly, the mean length of the JT was 1.35 +/- 0.15 cm (range 1-2.8 cm); the mean width, 1.10 +/- 0.12 cm (range 0.8-1.3 cm); and the mean thickness, 0.51 +/- 0.18 cm (range 0.2-1 cm). CONCLUSIONS: Morphometric data on the JT contribute significantly to the neurosurgeon's task of skull base drilling. The 3D CT scans were useful in the preoperative planning. PMID- 17121140 TI - Expanded polytetrafluoroethylene graft for bypass surgery using the excimer laser assisted nonocclusive anastomosis technique. AB - OBJECT: Patients with complex craniocerebral pathophysiologies such as giant cerebral aneurysms, skull base tumors, and/or carotid artery occlusive disease are candidates for a revascularization procedure to augment or preserve cerebral blood flow. However, the brain is susceptible to ischemia, and therefore the excimer laser-assisted nonocclusive anastomosis (ELANA) technique has been developed to overcome temporary occlusion. Harvesting autologous vessels of reasonable quality, which is necessary for this technique, may at times be problematic or impossible due to the underlying systemic vascular disease. The use of artificial vessels is therefore an alternative graft for revascularization. Note, however, that it is unknown to what degree these grafts are subject to occlusion using the ELANA anastomosis technique. Therefore, the authors studied the ELANA technique in combination with an expanded polytetrafluoroethylene (ePTFE) graft. METHODS: The experimental surgeries involved bypassing the abdominal aorta in the rabbit. Ten rabbits were subjected to operations representing 20 ePTFE graft-ELANA end-to-side anastomoses. Intraoperative blood flow, followup angiograms, and long-term histological characteristics were assessed 75, 125, and 180 days postoperatively. Angiography results proved long-term patency of ePTFE grafts in all animals at all time points studied. Data from the histological analysis showed minimal intimal reaction at the anastomosis site up to 180 days postoperatively. Endothelialization of the ePTFE graft was progressive over time. CONCLUSIONS: The ELANA technique in combination with the ePTFE graft seems to have favorable attributes for end-to-side anastomoses and may be suitable for bypass procedures. PMID- 17121141 TI - Photophobia in a blind patient: An alternate visual pathway. Case report. AB - Photophobia is a common neurological and ophthalmological symptom that has been associated with a growing number of neurosurgical conditions, especially compressive lesions. The exact signaling pathways and neurophysiological features of the disorder are not well understood; however, data from multiple studies have shown the significance of the trigeminal system and the pretectal nuclei in its pathophysiology. The authors report on a rare case of a blind patient who presented with photophobia without evidence of light perception. They also review the literature and early experimental data in an effort to understand the possible neuronal pathways and structures involved in photophobia. PMID- 17121142 TI - Spontaneous resolution of syringomyelia and Chiari malformation Type I in a patient with cerebrospinal fluid otorrhea. Case report. AB - The spontaneous resolution of syringomyelia in the setting of a Chiari malformation Type I (CM-I) has been reported infrequently. Several theories about the pathogenesis and spontaneous resolution of syringomyelia associated with CM-I have been proposed. The authors present the case of a patient with spontaneous resolution of a CM-I and syringomyelia coinciding with the development of cerebrospinal fluid (CSF) otorrhea. Although cases of spontaneous resolution of syringomyelia have been reported, this is the first reported case of spontaneous resolution of syringomyelia and a CM-I associated with the simultaneous development of CSF otorrhea. PMID- 17121143 TI - Postoperative dural sinus thrombosis in a patient in a hypercoagulable state. Case report. AB - Spontaneous cerebral venous sinus thrombosis is a rare problem that may be encountered in patients with underlying thrombophilic disorders. It has also been reported as a postoperative complication following suboccipital, transpetrosal, and transcallosal approaches. The authors report on a 67-year-old man with two prior episodes of lower-extremity deep venous thrombosis who underwent transcallosal resection of a colloid cyst and in whom sagittal sinus thrombosis developed 2 weeks thereafter. Results of a subsequent hematological workup revealed both a factor V Leiden mutation and the presence of antiphospholipid antibodies, two thrombophilic risk factors that likely contributed to the development of delayed postoperative sinus thrombosis. Although the safety of low molecular-weight heparin (LMWH) after craniotomy has not been established in a randomized, controlled study, there is sufficient evidence to justify its use for prophylactic anticoagulation therapy in patients at high risk for postoperative cerebral venous thrombosis. The authors propose using LMWH prophylaxis in patients with thrombophilic disorders who undergo neurosurgical procedures in proximity to dural sinuses in an effort to prevent catastrophic venous infarction. PMID- 17121144 TI - Stereotactic radiosurgery as adjuvant treatment for residual solitary fibrous tumor. Case report. AB - This 50-year-old woman presented with headache and visual disturbance. Neuroimaging results demonstrated a well-demarcated tumor attached to the falx cerebri near the transverse sinus, and the lesion was subtotally removed. Based on histological and immunohistochemical evaluation, a solitary fibrous tumor (SFT) was diagnosed. During the next 32 months, the size of the residual tumor increased slightly at the transverse sinus. Stereotactic radiosurgery (SRS) was performed as an adjuvant treatment. Over the course of the next 4 years the tumor decreased in size. The authors suggest that SRS constitutes good adjuvant treatment for regrowing residual SFTs not amenable to reoperation. PMID- 17121145 TI - Endovascular surgery for very small ruptured intracranial aneurysms. Technical note. AB - Application of endovascular surgery for very small aneurysms is controversial because of technical difficulties and high complication rates. The aim in the present study was to assess treatment results in a series of such lesions at one institution. Since 1997, endovascular surgery has been advocated for very small ruptured aneurysms (< 3 mm in maximum diameter) that fulfill the criterion of a fundus/neck ratio greater than 1.5. Twenty-one patients were treated, for whom the World Federation of Neurosurgical Societies classification before treatment was Grade I in 10, Grade II in two, Grade III in two, Grade IV in five, and Grade V in two. The aneurysm location was the internal carotid artery in four, the anterior communicating artery in 11, the middle cerebral artery in one, and the vertebrobasilar system in five. In all patients, endovascular surgery was performed using Guglielmi detachable coils after induction of general anesthesia. Initially, the presumed volume of the lesions was calculated for each aneurysm. Thereafter, the appropriate coil length was decided according to the volume embolization ratio, as 30 to 40%. In all attempts to obliterate aneurysms a single coil was used. All aneurysms were completely obliterated as confirmed by postembolization angiography, without procedure-related complications. During the follow-up period only one patient needed additional coil embolization for a growing aneurysm. Final outcomes were good recovery in 15 patients, moderate disability in five, and severe disability in one. Appropriate selection of patients and coils, and use of sophisticated techniques allow a good outcome for patients with very small aneurysms. PMID- 17121146 TI - Treatment of vertebral artery aneurysms with transposition of the posterior inferior cerebellar artery to the vertebral artery combined with parent artery occlusion. Technical note. AB - The authors describe transposition of the posterior inferior cerebellar artery (PICA) to the vertebral artery (VA) combined with parent artery occlusion for the treatment of VA aneurysms in cases in which a clip could not be applied because of the origin of the ipsilateral PICA. The aneurysm is trapped through a lower lateral suboccipital craniectomy. The PICA is then cut just distal to the aneurysm, and the PICA and VA proximal to the aneurysm are anastomosed in an end to-end or end-to-side fashion. The surgical procedure was successfully performed in two patients, each of whom had hypoplastic occipital arteries (OAs). The PICA contralateral to the lesion was hypoplastic in one patient and distant to the ipsilateral PICA in the other patient. Mild transient dysphagia developed postoperatively in one patient due to glossopharyngeal and vagus nerve palsy, and the other patient had an uneventful postoperative course. In both patients, postoperative cerebral angiography demonstrated good patency of the transposed PICA. These results show that transposition of the PICA to the VA is a useful procedure for the reconstruction of the PICA when parent artery occlusion is necessary to exclude a VA aneurysm involving the origin of the PICA and when OA PICA anastomosis or PICA-PICA anastomosis cannot be performed. PMID- 17121147 TI - Wrap-clipping with polytetrafluoroethylene for ruptured blisterlike aneurysms of the internal carotid artery. Technical note. AB - A technique combining wrapping and clip occlusion of aneurysms by using polytetrafluoroethylene (PTFE) for treatment of ruptured blisterlike aneurysms of the supraclinoid internal carotid artery (ICA) is described. The diameter of the abnormal arterial lesion along the long axis of the ICA and the distance between the origin of the ophthalmic artery and the origin of the posterior communicating artery (PCoA), or the origin of the PCoA and the origin of the anterior choroidal artery are measured intraoperatively; a strip of PTFE membrane is then trimmed with scissors to match this diameter and distance. After temporarily occluding the cervical ICA, the intracranial ICA that includes the lesion is wrapped with the strip of PTFE, and one or more aneurysm clips are applied parallel to the ICA. This procedure was successfully accomplished in six patients, all of whom had an uneventful postoperative course with no recurrent subarachnoid hemorrhage during the follow-up period. "Wrap-clipping" using PTFE is a useful procedure for management of ruptured blisterlike aneurysms of the ICA. PMID- 17121148 TI - Spontaneous thrombosis and resorption of a giant fusiform A2 aneurysm. Case illustration. PMID- 17121149 TI - Neuroanatomy and cadaver dissection in Italy: History, medicolegal issues, and neurosurgical perspectives. AB - Despite the significant Italian tradition of important anatomical studies, an outdated law historically influenced by the Catholic church restricts the use of cadavers for teaching and scientific purposes. The object of the present paper was to trace the historical evolution of the Italian anatomical tradition, particularly neuroanatomical studies, in relation to the juridical regulations on the use of cadavers today. Special attention was paid to the opportunities offered to neurosurgery by using cadavers and to the scientific and social issues in neurosurgical training in the twenty-first century. Considering the new Common European Constitution, the authors advocate a political solution from the European community to improve the quality of training in the disciplines with a social impact such as neurosurgery. PMID- 17121150 TI - Embolization of orbital arteriovenous fistulas. PMID- 17121151 TI - Modeling the risk of unruptured intracranial aneurysms and the benefit of prophylactic treatment. PMID- 17121152 TI - Cystic intracranial plasma cell granuloma. PMID- 17121153 TI - Idiopathic normal-pressure hydrocephalus. PMID- 17121154 TI - Neuronavigation in solitary intracerebral tumors. PMID- 17121155 TI - Intracranial aneurysms and antihypertension therapy. PMID- 17121156 TI - Myoblast therapy: from bench to bedside. AB - Myoblasts are defined as stem cells containing skeletal muscle cell precursors. A decade of experimental work has revealed many properties of myoblasts, including the stability of resulting hybrid myofibers without immune suppression, the persistence of transgene expression, and the lack of tumorigenicity. Early phase clinical trials also showed that myoblast-based therapy is a promising approach for many intractable clinical conditions, including both muscle-related and non muscle-related diseases. The potential application of myoblast therapy may be in the treatment of genetic muscle diseases, cardiomyocyte damaged heart diseases, and urinary incontinence. This review will provide an overview of myoblast biology, along with discussion of the potential application in clinical medicine. In addition, problems in current myoblast therapy and possible future improvements will be addressed. PMID- 17121157 TI - Cell therapy for Parkinson's disease: only young onset patients allowed? Reflections about the results of recent clinical trials with cell therapy and the progression of Parkinson's disease. AB - The selection of the best candidates for surgery among Parkinson's disease (PD) patients is a debated topic. This could be particularly important for transplantation studies in which patients with advanced PD and motor complications refractory to conventional pharmacological treatments are usually included. The development of lesions in nondopaminergic structures, which apparently are unaffected by the intervention, could eventually lead to the appearance of disabling, treatment-resistant symptoms. This has been considered as the crucial factor responsible for the outcome of any therapeutic procedure. However, other factors might be involved. It is suggested in this article that the rate of progression of PD and the effects of ageing are more important than the extradopaminergic involvement in the final outcome. Rate of progression of PD is critically related to the power of compensatory mechanisms, which are age related and under the control of still unknown genes. Thus, patients with young onset parkinsonism (YOP), either caused by gene mutations or not, could be the best candidates for surgery because they have a slower disease progression and more competent compensatory mechanisms. On the other hand, this can also explain the appearance of unexpected side effects such as the "runaway" dyskinesias reported following transplantation. PMID- 17121158 TI - Bridging nigrostriatal pathway with fibroblast growth factor-primed peripheral nerves and fetal ventral mesencephalon transplant recuperates from deficits in parkinsonian rats. AB - Previous studies have indicated that the nigrostriatal dopaminergic (DA) pathway can be reconstructed in hemiparkinsonian rats with a bridge transplantation technique involving fetal ventral mesencephalic transplants and glial cell line derived neurotrophic factor. In this study, we examined if the nigrostriatal pathway can be restored by combining peripheral nervous tissue with the fetal ventral mesencephalon transplants. Adult rats were injected with 6 hydroxydopamine into left median forebrain bundle. Those with marked rotational behavior, which has been previously shown to indicate complete DA dennervtion, were used for transplant treatments. One month after the lesion, fetal ventral mesencephalic cells were transplanted into the nigral region followed by nigral striatal grafting of peripheral nerves as a bridge. The bridging nerves (sciatic or intercostals) were pretreated with basic fibrous growth factor (nerve+bFGF+) or Hank's saline (nerve+bFGF-). We found that (a) animals receiving transplants of VM and bFGF+ nerve had a reduction in rotational behavior; (b) animals receiving bFGF-- nerve bridge only had a partial improvement in rotation. Reinnervation of tyrosine hydroxylase (TH)-immunoreactive (ir) fibers into the striatum was found in both of the above groups with more innervation in the former than in the latter. No TH-ir fibers in lesioned striatum or reduction in rotational behavior were found in animals receiving VM only, or VM plus bFGF. Taken together, our data indicate that peripheral nerve, with the aid of bFGF, greatly facilitates the reconstitution of the TH pathway from nigra to striatum and improves motor function in hemiparkinsonian rats. PMID- 17121159 TI - Inhibition of p38 MAP kinase in the early posttransplantation phase redistributes blood vessels from the surrounding stroma into the transplanted endocrine tissue. AB - Transplanted pancreatic islets attain a chronically decreased vascular density following transplantation, despite the increased concentrations of vascular endothelial growth factor (VEGF) secreted from beta-cells in response to hypoxia during culture and in the immediate posttransplantation phase. VEGF, however, exerts dual effects on endothelial cells, and in islet endothelial cells of the adult, the vascular permeability-inducing effects of VEGF seem normally more pronounced than those to induce angiogenesis. p38 MAP kinase activity has recently been shown to serve as a switch to separate these properties of VEGF; inhibition of p38 MAP kinase activity enhances VEGF-induced angiogenesis and, at the same time, abrogates VEGF-induced vascular permeability. We hypothesized that the revascularization of transplanted islets may be hampered by a predisposition of adult islet endothelial cells to react to VEGF by forming fenestrae rather than migrating and proliferating. We therefore administered the p38 MAP kinase inhibitor SB203580 by daily IP injections for the first 14 days following transplantation, and then studied the influence of this treatment on the oxygen tension, blood perfusion, and vascular density of the islet grafts 1 month posttransplantation. SB203580 treatment redistributed islet graft blood vessels from the stroma into the endocrine tissue, and this redistribution of blood vessels into the endocrine tissue was accompanied by an increased oxygenation of the islet cells. However, the total number of blood vessels in the tissue was not affected. The blood perfusion of the islet grafts was also similar in control and SB203580-treated animals. Our results suggest that effects of VEGF to preferentially induce vascular permeability may partially contribute to, but is not the main cause of, low revascularization of transplanted islets. PMID- 17121160 TI - Isolated pancreatic islets in three-dimensional matrices are responsive to stimulators and inhibitors of angiogenesis. AB - The formation of a new microvasculature is essential for the long-term survival and function of the islet graft. In this study we examined endothelium of isolated pancreatic islets by stimulation with growth factors, different culture conditions, and genetic modification. We also inspected the effect of immunosuppressives used in human transplantation on angiogenesis. Isolated islets were embedded in a three-dimensional fibrin or Matrigel matrix. The effect of hyperglycemia, hypoxia, and the addition of VEGF and bFGF was investigated. We exposed islets from transgenic mice expressing the VEGF gene (RIP1VEGF-A) to high glucose (16.7 mmol/L) medium and tested the immunosuppressive agents rapamycin (100 ng/ml) and FK506 (100 ng/ml). To quantify angiogenesis the percentage of sprouting islets was determined. New endothelial capillary-like structures protruded from isolated pancreatic islets. Addition of VEGF to the islets and transgenic RIP-VEGF islets showed a two- to threefold increase of sprouting islets compared to control. Hypoxic culture conditions stimulated angiogenesis, resulting in a twofold increase of capillary sprouting. Rapamycin and FK506 proved to be potent inhibitors of angiogenesis in this system, because a decrease of sprouting islets of more than 20% by both agents was observed. Isolated pancreatic islets are capable of forming new capillary structures and are susceptible to pro- and antiangiogenic stimuli. PMID- 17121161 TI - Endothelial cell preservation at 10 degrees C minimizes catalytic iron, oxidative stress, and cold-induced injury. AB - There is growing evidence that oxidative stress plays an important role in mediating the injury induced by hypothermia during the preservation of cells and tissues for clinical or research use. In cardiovascular allografts, endothelial cell loss or injury may lead to impaired control of vascular permeability and tone, thrombosis, and inflammation. We hypothesized that hypothermia-induced damage to the endothelium is linked to increases in intracellular catalytic iron pools and oxidative stress. In this study, bovine aortic endothelial cells and cell culture methods were used to model the response of the endothelium of cardiovascular tissues to hypothermia. Confluent cells were stored at 0 degrees C to 25 degrees C and cell damage was measured by lipid peroxidation (LPO) and lactate dehydrogenase release. Varying the bleomycin-detectible iron (BDI) in cells modulated cold-induced LPO and cell injury. In untreated cells, injury was highest at 0 degrees C and a minimum at 10 degrees C. A similar temperature dependent trend was found in BDI levels and cell plating efficiencies. Arrhenius plots of cell killing and iron accumulation rates showed biphasic temperature dependence, with minima at 10 degrees C and matching activation energies above and below 10 degrees C. These findings imply that the mechanisms underlying the hypothermic increase in catalytic iron, oxidative stress, and cell killing are the same and that preservation of the endothelium may be optimized at temperatures above those routinely used. PMID- 17121162 TI - Application of a new subretinal injection device in the dog. AB - The use of a new subretinal injection device (RetinaJect Subretinal Cannula, SurModics, Inc., Eden Prairie, MN) to access the subretinal space in the canine model was evaluated. Subretinal injections were performed in 33 mongrel dogs between 2 and 52 months of age (median = 9 months). In 5 normal dogs the injection of 150 microl saline or India ink occurred by using a conventional subretinal injection device (CSID) with a 30-gauge anterior chamber irrigating cannula. The sclera had to be surgically exposed and penetrated before the subretinal injection with the CSID could occur. After removing the CSID, the conjunctiva over the sclerotomy site had to be closed. In a second group of 28 dogs [16 normals, 10 RPE65 mutants, and 2 with progressive rod cone degeneration (prcd)], the 25-gauge needle of the RetinaJect was used to penetrate the conjunctiva and the sclera. Once the tip of the needle was close to the retinal surface, a 39-gauge polyimide cannula was extended and brought into apposition with the retina for the subsequent subretinal injection of 150 microl saline, India ink, or adeno-associated virus (AAV). No closure of the conjunctiva was required. The animals were clinically monitored between 1 and 59 weeks after surgery. From this second group 25 eyes were harvested for routine histological analysis either immediately after surgery or after a clinical observation time of between 1 and 40 weeks. Both devices provided equally successful access to the subretinal space. The main advantage of the RetinaJect was that no surgical dissection was required; this led to a shorter procedure time and milder postoperative conjunctival swelling. In summary, the use of the RetinaJect can be recommended as an alternative to the CSID for subretinal injections in dogs. PMID- 17121163 TI - Periosteal cell pellet culture system: a new technique for bone engineering. AB - To treat bone loss that is induced by disease or wounds, bone grafts are commonly used. In dentistry, guided tissue regeneration is effective in the treatment of periodontal diseases. However, bone resorption after implantation is a major problem with the bone graft and guided tissue regeneration technique. This study examines a cell pellet culture system without exogenous scaffolds for bone regeneration. First, we examined the effect of ascorbic acid on cells. Transmission electron microscopic observation revealed that cells formed a three dimensional structure of multiple cell layers after 5 weeks of culturing in medium containing 50 microg/ ml ascorbic acid with the medium changed every 7 days. A single cell pellet was produced by centrifuging cells that were gathered from 10 tissue culture dishes. Van Gieson staining and collagen type I immunostaining showed that the pellet contained collagen fibers and cells that adhered to the collagen fibers. Several of these cell pellets were implanted subcutaneously on the backs of nude mice for 6 weeks. Histology and immunohistochemistry results indicated new bone formation, vascular invasion, and insular areas of calcification. Bone tissue was surrounded by osteoblasts. The appearance of new bone formation is similar to that seen in intramembranous ossification. The present pellet system is reliable and might solve problems of bone resorption after implantation. PMID- 17121164 TI - Directed three-dimensional growth of microvascular cells and isolated microvessel fragments. AB - Tissue engineering has promise as a means for repairing diseased and damaged tissues. A significant challenge in tissue construction relates to the constraints placed on tissue geometries resulting from diffusion limitations. An ability to incorporate a premade vasculature would overcome these difficulties and promote construct viability once implanted. Most in vitro microvascular fabrication strategies rely on surface-associated cell growth, manipulated cell monolayers, or random arrangement of cells within matrix materials. In contrast, we successfully suspended microvascular cells and isolated microvessel fragments within collagen and then microfluidically drove the mixtures into microfabricated network topologies. Developing within the 3D collagen matrix, patterned cells progressed into cord-like morphologies. These geometries were directed by the surrounding elastomer mold. With similar techniques, suspended fragments formed endothelial sprouts. By avoiding the addition of exogenous growth factors, we allowed constituent cells and fragments to autonomously develop within the constructs, providing a more physiologically relevant system for in vitro microvascular development. In addition, we present the first examples of directed endothelial cell sprouting from parent microvessel fragments. We believe this system may serve as a foundation for future in vivo fabrication of microvascular networks for tissue engineering applications. PMID- 17121165 TI - Community treatment orders for psychiatric patients: the emperor with no clothes. PMID- 17121166 TI - Community treatment orders: an essential therapeutic tool in the face of continuing deinstitutionalization. PMID- 17121167 TI - Psychopathy checklist score predicts negative events during the sentences of prisoners with Hare psychopathy: a prospective study at a German prison. AB - OBJECTIVE: This study examined the predictive validity of the German translation of the Psychopathy Checklist-Screening Version (PCL-SV) for negative events during the course of the prison sentence of German prisoners. METHOD: Using the PCL-SV, we investigated 145 offenders in a German prison at the start of their sentences. We then compared the extreme groups identified by the PCL-SV--the high and low scorers--using a prospective design with respect to negative events and factors during the course of the sentences. This involved the standardized collection of data on both objective records of disciplinary incidents and subjective impressions from prison staff on the basis of operationalized criteria. RESULTS: The high scorers were involved in significantly more disciplinary incidents and were also rated significantly less favourably by prison staff than the low scorers. CONCLUSION: Until now, the PCL has only been shown to predict recidivism following release from prison. The results of our study show that the PCL also has predictive validity for problems during the course of the sentence. It is therefore recommended that the PCL be used routinely at the start of the prison sentence to estimate the likelihood of subsequent difficulties. PMID- 17121168 TI - The associations between peer and parental relationships and suicidal behaviours in early adolescents. AB - OBJECTIVE: To investigate associations between suicidal behaviours, including suicidal ideation and attempts, and poor peer and parental relationships in a nationally representative sample of Canadian adolescents aged 12 to 13 years. METHODS: We used Statistics Canada's National Longitudinal Survey of Children and Youth as the dataset. This cross-sectional sample included 1049 girls and 1041 boys aged 12 to 13 years. We obtained answers to self-report questionnaires that included measures of peer relationships, parental nurturance, and parental rejection, as well as information regarding suicidal ideation and attempts. Unadjusted and adjusted logistic regression models were used for the analysis. We included depression in the multiple logistic regression analysis. Analyses were run separately for boys and girls. RESULTS: The unadjusted logistic regression models found that, among early adolescent boys and girls, depression, poor peer relationships, decreased parental nurturance, and increased parental rejection were all significantly associated with suicidal ideation and attempts. However, after adjusting for all other variables in the multiple logistic regression models, poor peer relationships were no longer significantly associated with suicidal ideation among early adolescent boys and were only weakly associated among early adolescent girls. CONCLUSIONS: Poor parental relationships and depression were more powerfully associated with suicidal ideation and attempts than were peer relationships in a nationally representative sample of boys and girls aged 12 to 13 years, and these factors may be important early intervention targets. PMID- 17121169 TI - Demographic characteristics of participants in studies of risk factors, prevention, and treatment of postpartum depression. AB - OBJECTIVES: Metaanalyses have found that sociodemographic variables are not strong predictors of postpartum depression. However, no studies have systematically examined the extent to which the samples used in published research on postpartum depression have included sufficiently diverse samples of women to merit this conclusion. The objectives o this study were to examine the demographic characteristics of participants in previously published studies and to document existing gaps in the current literature. METHOD: We extracted age, ethnicity, relationship status, and socioeconomic status of 51 453 participants from 143 studies previously selected for systematic literature reviews. RESULTS: Few studies reported complete demographic data; however, existing data indicate that participants were predominantly aged 25 to 35 years, white, partnered, and of mid- or high-socioeconomic status. CONCLUSIONS: To assess the external validity of the findings, improved reporting of demographic characteristics is required in publications related to postpartum depression. Additional research is needed to understand postpartum depression among understudied populations. PMID- 17121170 TI - One-year stability of diagnosis in first-episode nonaffective psychosis: influence of sex. AB - OBJECTIVE: To determine whether sex influences the frequency and direction of diagnostic change 1 year after a first episode of nonaffective psychosis. METHOD: The subjects (n = 228) were individuals diagnosed with schizophrenia and other psychoses according to DSM-IV criteria at baseline and at 1 year. RESULTS: When diagnostic shifts occurred, men mainly shifted toward schizophrenia (84.6%), which was associated with more severe symptoms. In contrast, 72% of women in whom a diagnostic shift occurred shifted away from a schizophrenia spectrum disorder toward mood disorder. A shift to mood disorder in women was associated with an early age of illness onset. CONCLUSIONS: At initial presentation, nonaffective first-episode psychosis studies may underestimate the number of men and overestimate the number of women who will eventually receive a schizophrenia diagnosis. PMID- 17121171 TI - Serum monitoring of antipsychotic drug levels during concomitant administration of sertraline and antipsychotic medication. AB - OBJECTIVE: To assess whether pharmacokinetic drug interactions occur when sertraline is added to antipsychotic medications. METHOD: Forty-eight patients with remitted DSM-IV schizophrenia and comorbid major depression were randomized to placebo for 6 weeks or sertraline 50 mg for 4 weeks followed by sertraline 50 mg to 100 mg for 2 weeks for nonresponders. Treatment with the patients' usual antipsychotic continued. Weekly clinical outcome assessments occurred for 6 weeks, and serum samples for drug monitoring were collected at Weeks 1, 5, and 6. Serum concentrations of sertraline and antipsychotics were measured with standard assays. RESULTS: In both placebo- and sertraline-treated groups, most patients displayed minor fluctuations in antipsychotic serum levels over 6 weeks. There was no clinical evidence of drug interactions in the sertraline-treated group. CONCLUSIONS: Clinically significant adverse effects did not occur despite variable antipsychotic serum levels with or without sertraline. Concern about pharmacokinetic interactions should not deter the use of sertraline for depression in individuals with schizophrenia. PMID- 17121172 TI - Forgetfulness: a role in noncompliance with antidepressant treatment. AB - OBJECTIVES: To determine the degree of noncompliance with antidepressant treatment in the Alberta population and to investigate the reasons for noncompliance. METHOD: We used data from the Alberta Mental Health Survey, a telephone survey conducted in 2003 (n = 5323 adults), to produce population-based estimates of the frequency of noncompliance and the reported reasons for noncompliance. RESULTS: Reported noncompliance was 41.7% (95% confidence interval [CI], 36.9% to 46.6%) for respondents taking 1, 2, or 3 antidepressants. Noncompliance for those taking 1 antidepressant was 42.0% (95%CI, 36.9% to 47.2%), whereas noncompliance for those taking 2 or 3 antidepressants was 39.4% (95%CI, 26.7% to 53.6%). Among respondents currently taking at least one antidepressant, 64.9% (95%CI, 57.4% to 71.7%) reported that forgetfulness was the most common reason for noncompliance. Of respondents taking 1 medication, 64.1% (95%CI, 56.0% to 71.4%) reported forgetfulness as did 71.3% (95%CI, 48.3% to 86.8%) of those taking 2 or 3 medications. Both the frequency of noncompliance and the reported reasons for noncompliance were independent of sex and age. CONCLUSION: Our study replicates prior reports that indicate that noncompliance is common with antidepressant treatment. Forgetting to take medication is the most important reported reason for this noncompliance. PMID- 17121173 TI - A new monoterpene glycoside and antibacterial monoterpene glycosides from Paeonia suffruticosa. AB - Antibacterial activity-guided fractionation of the CHCI3-MeOH (1:1) extract of Paeonia suffruticosa root bark furnished three monoterpene glycosides, 6-O vanillyoxypaeoniflorin (1), mudanpioside-H (2), and galloyl-oxypaeoniflorin (3). Of the isolated compounds, compound 1 is a new compound. All isolated compounds showed broad, but moderate, antibacterial activity with minimum inhibitory concentration (MIC) values in the range of 100 to 500 microg/mL against eighteen pathogenic microorganisms of concern for public health or zoonosis. PMID- 17121174 TI - Constituents of the roots of Pueraria lobata inhibit formation of advanced glycation end products (AGEs). AB - Two isoflavone C-glucosides, puerarin (1) and PG-3 (2), a but-2-enolide, (+/-) puerol B (3), two isoflavone O-glucosides, daidzin (4) and genistin (5), and three pterocarpans, (-)-medicarpin (6), (-)-glycinol (7) and (-)-tuberosin (8), were isolated from a MeOH extract of the roots of Pueraria lobata, using an in vitro bioassay based on the inhibition of the formation of advanced glycation end products (AGEs) to monitor chromatographic fractionation. The structures of 1-8 were determined by spectroscopic data interpretation, particularly by 1D- and 2D NMR studies, and by comparison of these data with values in the literature. All of the isolates (1-8) were evaluated for their inhibitory activity on AGEs formation in vitro. Of these, puerarin (1), PG-3 (2), and (+/-)-puerol B (3) exhibited more potent inhibitory activity than the positive control aminoguanidine. PMID- 17121175 TI - Synthesis of novel benzofuran and related benzimidazole derivatives for evaluation of in vitro anti-HIV-1, anticancer and antimicrobial activities. AB - Previously, we synthesized and evaluated several benzofuran derivatives containing heterocyclic ring substituents linked to the benzofuran nucleus at C-2 by a two- to four-atom spacer as potential anti-HIV-1, anticancer and antimicrobial agents. Among these derivatives, NSC 725612 and NSC 725716 exhibited interesting anti-HIV-1 activity. To further investigate the structure activity relationship, we synthesized several new benzofuran derivatives derived from 2-acetylbenzofuran (2, 3a-c) and 2-bromoacetylbenzofuran (6; 7a,b; 8a,b). The compounds were designed to comprise the heterocyclic substituents directly linked to the benzofuran nucleus at C-2. Moreover, various related benzimidazoles derived from 2-acetylbenzimidazole and from 2-cyanomethylbenzimidazole (12a,b; 13a,b; 15; 16a,b) were also prepared as isosteres. The synthesized compounds were preliminarily evaluated for their in vitro anti-HIV-1, anticancer and antimicrobial activity. Compounds 2, 3a, 3b, and 12b showed weak anti-HIV-1 activity. Compound 6 exhibited mild activity against S. aureus, while compound 15 had mild activity towards S. aureus and C. albicans. However, no significant anticancer activity was observed with any of the tested compounds. From these results, we conclude that the presence of the spacer between the heterocyclic substituent and the benzofuran nucleus may be essential for the biological activity. PMID- 17121176 TI - Torilin from Torilis japonica (Houtt.) DC. blocks hKv1.5 channel current. AB - Torilin was purified from Torilis japonica (Houtt.) DC., and its effects on a rapidly activating delayed rectifier K+ channel (hKv1.5), cloned from human heart and stably expressed in Ltk- cells, as well as the corresponding K+ current (the ultrarapid delayed rectifier, I(KUR)) were assessed in human atrial myocytes. Using the whole cell configuration of the patch-clamp technique, torilin was found to inhibit the hKv1.5 current in time and voltage-dependent manners, with an IC50 value of 2.51+/-0.34 microM at +60 mV. Torilin accelerated the inactivation kinetics of the hKv1.5 channel, and slowed the deactivation kinetics of the hKv1.5 current, resulting in a tail crossover phenomenon. Additionally, torilin inhibited the hKv1.5 current in a use-dependent manner. These results strongly suggest that torilin is a type of open-channel blocker of the hKv1.5 channel. PMID- 17121177 TI - Exploration of essential structure of malloapelta B for the inhibitory activity against TNF induced NF-kappaB activation. AB - For the exploration of pharmacophoric moiety of malloapelta B (1) possessing the inhibitory activity of NF-kappaB activation, structural variation of alpha,beta unsaturated carbonyl motif was attempted. 1 was reduced by catalytic hydrogenation, sodium borohydride, and lithium aluminumhydride. Catalytic hydrogenation with 30 psi or 15 psi of H2 gas of 1 generated 8-butyl-5,7 dimethoxy-2,2-dimethylchroman (2) and 1-(5,7-dimethoxy-2,2-dimethylchroman-8 yl)butan-1-one (3), respectively. Reduction with sodium borohydride occurred at the double bond of alpha,beta-unsaturated ketone of 1 to give 1-(5,7-dimethoxy 2,2-dimethyl-2H-chromen-8-yl)butan-1-one (4). Reduction of 1 with lithium aluminumhydride and then quenched with methanol and water produced unexpected products, 1-(5,7-dimethoxy-2,2-dimethyl-2H-chromen-8-yl)-3-methoxy-1-butene (5) and 1-(5,7-dimethoxy-2,2-dimethyl-2H-chromen-8-yl)-3-hydroxy-l-butene (6). These are formed from the isomerization of initial product 9 through the continuous conjugate carbocation intermediate 11. Addition of ethylmagnesium bromide and dimethyl malonate anion to 1 gave the conjugate adducts 7 and 8. Ethylmagesium bromide and sodium borohydride reduction unusually gave the conjugate addition due to steric congestion around carbonyl group of 1. Compound 2 exhibits the reduced inhibitory activity against NF-kappaB activation and the others do not show the activity. Therefore alpha,beta-unsaturated carbonyl group of 1 should be important for its inhibitory activity. PMID- 17121178 TI - Cytotoxic terpene hydroperoxides from the aerial parts of Aster spathulifolius. AB - Three new sesquiterpene hydroperoxides, 1-[3-(2-hydroperoxy-3-methylbut-3-en)-4 hydroxyphenyl]ethanone (2), 7beta-hydroperoxy-eudesma-11-en-4-ol (3), and 7alpha hydroperoxymanool (4), together with three known compounds, germacrone (1), ent germacra-4(15),5,10(14)-trien-1alpha-ol (5) and teucdiol A (6) were isolated from the aerial parts of Aster spathulifolius (Compositae). Their structures were characterized using chemical and spectroscopic methods. The isolated compounds were tested for their cytotoxicity against five human tumor cell lines in vitro using a SRB method. The two new compounds, 3 and 4, showed moderate cytotoxicity against human cancer cells with ED50 values ranging from 0.24 to 13.27 microg/mL. PMID- 17121179 TI - Anti-inflammatory action of phenolic compounds from Gastrodia elata root. AB - Previous screening of the pharmacological action of Gastrodia elata (GE) root (Orchidaceae) showed that methanol (MeOH) extracts have significant anti inflammatory properties. The anti-inflammatory agents of GE, however, remain unclear. In this experiment, MeOH extracts of GE were fractionated with organic solvents for the anti-inflammatory activity-guided separation of GE. Eight phenolic compounds from the ether (EtOEt) and ethyl acetate (EtOAc) fractions were isolated by column chromatography: 4-hydroxybenzaldehyde (I), 4 hydroxybenzyl alcohol (II), benzyl alcohol (III), bis-(4-hydroxyphenyl) methane (IV), 4(4'-hydroxybenzyloxy)benzyl methylether (V), 4-hydroxy-3-methoxybenzyl alcohol (VI), 4-hydroxy-3-methoxybenzaldehyde (VII), and 4-hydroxy-3 methoxybenzoic acid (VIII). To investigate the anti-inflammatory and anti-oxidant activity of these compounds, their effects on carrageenan-induced paw edema, arachidonic acid (AA)-induced ear edema and analgesic activity in acetic acid (HAc)-induced writhing response were carried out in vivo; cyclooxygenase (COX) activity, reactive oxygen species (ROS) generation in rat basophilic leukemia (RBL 2H3) cells and 1,1-diphenyl-2-picryl-hydroazyl (DPPH) scavenging activity were determined in vitro. These phenolic compounds not only had anti-inflammatory and analgesic properties in vivo, but also inhibited COX activity and silica induced ROS generation in a dose-dependent manner. Among these phenolic compounds, compound VII was the most potent anti-inflammatory and analgesic. Compound VII significantly inhibited silica-induced ROS generation and compound VI significantly increased DPPH radical scavenging activity. Compounds I, II and III significantly inhibited the activity of COX-I and II. These results indicate that phenolic compounds of GE are anti-inflammatory, which may be related to inhibition of COX activity and to anti-oxidant activity. Consideration of the structure-activity relationship of the phenolic derivatives from GE on the anti inflammatory action revealed that both C-4 hydroxy and C-3 methoxy radicals of benzyl aldehyde play an important role in anti-inflammatory activities. PMID- 17121180 TI - The antitumor effects of selenium compound Na5SeV5O18.3H20 in K562 cell. AB - With an approach to study the anti-tumor effects and mechanism of selenium compound, we investigated the anti-tumor activity and mechanism of Na5SeV5O18.H20 (NaSeVO) in K562 cells. The results showed that 0.625-20 mg/L NaSeVO could significantly inhibit the proliferation of K562 cells in vitro in a time- and concentration-dependent manner as determined by microculture tetrazolium (MTT) assay, the IC50 values were 14.41 (4.45-46.60) and 3.45 (2.29-5.22) mg/L after 48 h and 72 h treatment with NaSeVO respectively. In vivo experiments demonstrated that i.p. administration of 5, 10 mg/kg NaSeVO exhibited an significant inhibitory effect on the growth of transplantation tumor sarcoma 180 (S180) and hepatoma 22 (H22) in mice, with inhibition rate 26.8% and 58.4% on S180 and 31.3% and 47.4% on H22, respectively. Cell cycle studies indicated that the proportion of G0/G1 phase was increased at 2.5 mg/ L while decreased at 10 mg/L after treatment for 24, 48 h. Whereas S phase was decreased at 2.5-5 mg/L and markedly increased at 10 mg/L after treatment for 48 h. After treatment for 24 h, 10 mg/L NaSeVO also markedly increased S and G2/M phases. Take together, the result clearly showed that NaSeVO markedly increased S and G2/M phases at 10 mg/L. The study of immunocytochemistry showed that the expression bcl-2 is significantly inhibited by 10 mg/L NaSeVO, and bax increased. Morphology observation also revealed typical apoptotic features. NaSeVO also significantly caused the accumulation of Ca2+ and Mg2+, reactive oxygen species (ROS) and the reduction of pH value and mitochondrial membrane potential in K562 cells as compared with control by confocal laser scanning microscope. These results suggest that NaSeVO has anti-tumor effects and its mechanism is attributed partially to apoptosis induced by the elevation of intracellular Ca2+, Mg2+ and ROS concentration, and a reduction of pH value and mitochondria membrane potential (MMP). PMID- 17121181 TI - Inhibitory effect of curcumin on MDR1 gene expression in patient leukemic cells. AB - When patients with cancers are treated with chemotherapeutic agents a long time, some of the cancer cells develop the multidrug resistance (MDR) phenotype. MDR cancer cells are characterized by the overexpression of multidrug resistance1(MDR1) gene which encodes P-glycoprotein (Pgp), a surface protein of tumor cells that functions to produce an excessive efflux and thereby an insufficient intracellular concentration of chemotherapeutic agents. A variety of studies have sought potent MDR modulators to decrease MDR1 gene expression in cancer cells. Our previous study has shown that curcumin exhibits characteristics of a MDR modulator in KB-V1 multidrug-resistant cells. The aim of this study was to further investigate the effect of curcumin on MDR1 gene expression in patient leukemic cells. The leukemic cells were collected from 78 childhood leukemia patients admitted at Maharaj Nakorn Chiang Mai Hospital, Chiang Mai, Thailand, in the period from July 2003 to February 2005. There were 61 cases of acute lymphoblastic leukemia (ALL), 14 cases of acute myeloblastic leukemia (AML), and 3 cases of chronic myelocytic leukemia (CML). There were 47 males and 31 females ranging from 1 to 15 years old. Bone marrows were collected. The leukemic cells were separated and cultured in the presence or absence of 10 microM curcumin for 48 hours. MDR1 mRNA levels were determined by RT-PCR. It was found that curcumin reduced MDR1 gene expression in the cells from 33 patients (42%). Curcumin affected the MDR1 gene expression in 5 of 11 relapsed cases (45%), 10 of 26 cases of drug maintenance (38%), 7 of 18 cases of completed treatment (39%), and 11 of 23 cases of new patients (48%). The expression levels of MDR1 gene in leukemic patient cells as compared to that of KB-V1 cells were classified as low level (1 20%) in 5 of 20 cases (25%), medium level (21-60%) in 14 of 32 cases (44%), and high level (61-100%) in 14 of 20 cases (70%). In summary, curcumin decreased MDR1 mRNA level in patient leukemic cells, especially in high level of MDR1 gene groups. Thus, curcumin treatment may provide a lead for clinical treatment of leukemia patients in the future. PMID- 17121182 TI - Effects of methyl gallate on arachidonic acid metabolizing enzymes: Cyclooxygenase-2 and 5-lipoxygenase in mouse bone marrow-derived mast cells. AB - Methyl gallate (MG) is a medicinal herbal product that is isolated from Paeonia lactiflora that inhibits cyclooxygenase-2 (COX-2) dependent phases of prostaglandin D2 (PGD2) generation in bone marrow-derived mast cells (BMMC) in a concentration-dependent manner with an IC50 values of 17.0 microM. This compound also found inhibited the COX-2-dependent conversion of the exogenous arachidonic acid to PGD2 in a dose-dependent manner with an IC50 values of 19.0 microM, using a COX enzyme assay kit. However, at concentrations up to 80 microM, MG did not inhibit COX-2 protein expression in BMMC, indicating that MG inhibits COX-2 activity directly. Furthermore, MG consistently inhibited the production of leukotriene C4 (LTC4) in a dose dependent manner, with an IC50 value of 5.3 microM. These results demonstrate that MG has a dual cyclooxygenase-2/5 lipoxygenase inhibitory activity, which might provide the basis for novel anti inflammatory drugs. PMID- 17121183 TI - Trials of novel 13C-urea-containing capsule for more economic and sensitive diagnosis of Helicobacter pylori infection in human subjects. AB - To develop a 13C-urea-containing capsule for more economic and sensitive diagnosis of Helicobacter pylori infection, the 13C-urea-containing capsules were prepared with various additives such as polyethylene glycol, microcrystalline cellulose, sodium lauryl sulfate and citric acid. Their dissolution test and 13C urea Breath Test in human volunteers were then performed. Polyethylene glycol increased the initial dissolution rates of urea and difference delta 13C values from 13C-urea, while microcrystalline cellulose and sodium lauryl sulfate decreased them. Irrespective of addition of citric acid, the compositions with polyethylene glycol showed higher values from 13C-urea compared to a commercial 76 mg 13C-urea-containing capsule due to higher initial dissolution rate. The capsules with 38 mg 13C-urea and 1.9 mg polyethylene glycol, which showed higher Helicobacter pylori-positive value of about 8 per thousand at 10 min, improved the sensitivity of 13C-urea in human volunteers. Thus, the 13C-urea-containing capsule with polyethylene glycol would be a more economical and sensitive preparation for diagnosis of Helicobacter pylori infection. PMID- 17121184 TI - Mutant presenilin 2 causes abnormality in the brain lipid profile in the development of Alzheimer's disease. AB - Mutation in the presenilin 2 (PS2mt) is known to be one of factors involved in the development of Alzheimer's disease (AD). It was recently revealed that an abnormality of lipid metabolism is a phenomenon occurring in AD. Therefore, the aim of this study was to investigate the potential relationship between the mutation of PS2 and alterations of the lipid profile within the brain. The results showed there increases in the levels of cholesterol, low density lipoprotein and triglyceride, but a decrease in the level of high density lipoprotein in brain tissues expressing mutant PS2. These findings indicated that PS2mt is involved in the abnormalities of the lipid profile, which could cause or result in the development of AD. PMID- 17121185 TI - Expression profiling of lipopolysaccharide target genes in RAW264.7 cells by oligonucleotide microarray analyses. AB - In inflammatory responses, induction of cytokines and other immune regulator genes in macrophages by pathogen-associated signal such as lipopolysaccharide (LPS) plays a crucial role. In this study, the gene expression profile changes by LPS treatment in the macrophage/monocyte lineage cell line RAW264.7 was investigated. A 60-mer oligonucleotide microarray of which probes target 32381 mouse genes was used. A reverse transcription-in vitro translation labeling protocol and a chemileuminescence detection system were employed. The mRNA expression levels in RAW264.7 cells treated for 6 h with LPS and the control vehicle were compared. 747 genes were up-regulated and 523 genes were down regulated by more than 2 folds. 320 genes showing more than 4-fold change by LPS treatment were further classified for the biological process, molecular function, and signaling pathway. The biological process categories that showed high number of increased genes include the immunity and defense, the nucleic acid metabolism, the protein metabolism and modification, and the signal transduction process. The chemokine-cytokine signaling, interleukin signaling, Toll receptor signaling, and apoptosis signaling pathways involved high number of genes differentially expressed in response to LPS. These expression profile data provide more comprehensive information on LPS-target genes in RAW264.7 cells, which will be useful in comparing gene expression changes induced by extracts and compounds from anti-inflammatory medicinal herbs. PMID- 17121186 TI - Antiplatelet and antithrombotic activities of Korean Red Ginseng. AB - The antiplatelet and antithrombotic activities of Korean Red Ginseng (KRG) were examined on rat carotid artery thrombosis in vivo, and platelet aggregation in vitro and ex vivo. Administration of KRG to rats not only prevented carotid artery thrombosis in vivo in a dose-dependent manner, but also significantly inhibited ADP- and collagen-induced platelet aggregation ex vivo, while failed to prolong coagulation times such as activated partial thromboplastin time (APTT) and prothrombin time (PT), indicating the antithrombotic effect of KRG might be due to its antiplatelet aggregation rather than anticoagulation effect. In line with the above observations, KRG inhibited U46619-, arachidonic acid-, collagen- and thrombin-induced rabbit platelet aggregation in vitro in a concentration dependent manner, with IC50 values of 620 +/- 12, 823 +/- 22, 722 + 21 and 650 +/ 14 microg/mL, respectively. Accordingly, KRG also inhibited various agonists induced platelet serotonin secretions as it suppressed platelet aggregation. These results suggest that KRG has a potent antithrombotic effect in vivo, which may be due to antiplatelet rather than anticoagulation activity, and KRG intake may be beneficial to the individuals with high risks of thrombotic and cardiovascular diseases. PMID- 17121187 TI - Inhibitory effects of paeonol on morphine-induced locomotor sensitization and conditioned place preference in mice. AB - The inhibitory effects of paeonol, a major compound of Paeoniae radix, on the development of locomotor sensitization, conditioned place preference (CPP) and dopamine receptor supersensitivity induced by the repeated administration of morphine were investigated through behavioral experiments. A single administration of morphine produces hyperlocomotion. Repeated administration of morphine develops sensitization (reverse tolerance), a progressive enhancement of locomotion, which is used as a model for studying the drug-induced drug-seeking behaviors, and CPP, which is used as a model for studying drug reinforcement. Paeonol inhibited morphine-induced hyperlocomotion, sensitization and CPP. In addition, paeonol inhibited the development of postsynaptic dopamine receptors supersensitivity, which may be an underlying common mechanism that mediates the morphine-induced dopaminergic behaviors such as sensitization and CPP. Apomorphine (a dopamine agonist)-induced climbing behaviors also were inhibited by a single direct administration of paeonol. These results provide evidence that paeonol exerts anti-dopaminergic activity, and it is suggested that paeonol may be useful for the prevention and therapy of these adverse actions of morphine. PMID- 17121188 TI - In vivo pharmacokinetics, activation of MAPK signaling and induction of phase II/III drug metabolizing enzymes/transporters by cancer chemopreventive compound BHA in the mice. AB - Phenolic antioxidant butylated hydroxyanisole (BHA) is a commonly used food preservative with broad biological activities, including protection against chemical-induced carcinogenesis, acute toxicity of chemicals, modulation of macromolecule synthesis and immune response, induction of phase II detoxifying enzymes, as well as its undesirable potential tumor-promoting activities. Understanding the molecular basis underlying these diverse biological actions of BHA is thus of great importance. Here we studied the pharmacokinetics, activation of signaling kinases and induction of phase II/III drug metabolizing enzymes/transporter gene expression by BHA in the mice. The peak plasma concentration of BHA achieved in our current study after oral administration of 200 mg/kg BHA was around 10 microM. This in vivo concentration might offer some insights for the many in vitro cell culture studies on signal transduction and induction of phase II genes using similar concentrations. The oral bioavailability (F) of BHA was about 43% in the mice. In the mouse liver, BHA induced the expression of phase II genes including NQO-1, HO-1, gamma-GCS, GST-pi and UGT 1A6, as well as some of the phase III transporter genes, such as MRP1 and Slcolb2. In addition, BHA activated distinct mitogen-activated protein kinases (MAPKs), c-Jun N-terminal kinase (JNK), extracellular signal-regulated protein kinase (ERK), as well as p38, suggesting that the MAPK pathways may play an important role in early signaling events leading to the regulation of gene expression including phase II drug metabolizing and some phase III drug transporter genes. This is the first study to demonstrate the in vivo pharmacokinetics of BHA, the in vivo activation of MAPK signaling proteins, as well as the in vivo induction of Phase II/III drug metabolizing enzymes/transporters in the mouse livers. PMID- 17121189 TI - Preparation of prolonged release clarithromycin microparticles for oral use and their in vitro evaluation. AB - Prolonged release microparticles of clarithromycin (CL) were prepared using Eudragit RL 100 and RS 100 by spray-drying and casting-drying techniques. For the characterization of those microparticles, preparation yield, particle size distribution, X-ray diffraction, thermal behavior, active agent content and in vitro dissolution from the microparticles were performed. HPLC was used for the assay of clarithromycin and the assay method was validated. All the formulations obtained showed prolonged release when compared to pure clarithromycin. Microparticles prepared by spray-drying method had a slower release compared to those of casting-drying method. Spray-drying method seems to be a more suitable method to prepare microparticles for prolongation in release. PMID- 17121190 TI - Enhanced transdermal delivery of pranoprofen from the bioadhesive gels. AB - Percutaneous delivery of NSAIDs has advantages of avoiding hepatic first pass effect and delivering the drug for extended period of time at a sustained, concentrated level at the inflammation site that mainly acts at the joint and the related regions. To develop the new topical formulations of pranoprofen that have suitable bioadhesion, the gel was formulated using hydroxypropyl methylcellulose (HPMC) and poloxamer 407. The effects of temperature on drug release was performed at 32 degrees C, 37 degrees C and 42 degrees C according to drug concentration of 0.04%, 0.08%, 0.12%, 0.16%, and 0.2% (w/w) using synthetic cellulose membrane at 37+/-0.5 degrees C. The increase of temperature showed the increased drug release. The activation energy (Ea), which were calculated from the slope of lop P versus 1000/T plots was 11.22 kcal/ mol for 0.04%, 10.79 kcal/mol for 0.08%, 10.41 kcal/mol for 0.12% and 8.88 kcal/mol for 0.16% loading dose from the pranoprofen gel. To increase the drug permeation, some kinds of penetration enhancers such as the ethylene glycols, the propylene glycols, the glycerides, the non-ionic surfactants and the fatty acids were incorporated in the gel formulation. Among the various enhancers used, propylene glycol mono laurate showed the highest enhancing effects with the enhancement factor of 2.74. The results of this study suggest that development of topical gel formulation of pranoprofen containing an enhancer is feasible. PMID- 17121191 TI - Effects of morin pretreatment on the pharmacokinetics of diltiazem and its major metabolite, desacetyldiltiazem in rats. PMID- 17121192 TI - Immediate polymerization shrinkage in light cured restorative resins. AB - The purpose of this study was to evaluate the immediate polymerization shrinkage of restorative resins following activation with different amounts of light energy. The composites used were: Z-100 (3M-ESPE), Z-250 (3M-ESPE), Ariston pHc (Ivoclar-Vivadent), Filtek P-60 (3M-ESPE), SureFil (Dentsply), Tetric Flow (Ivoclar-Vivadent), Silux Plus (3M-ESPE) and F-2000 (3M-ESPE). The percentage shrinkage after an 80-second activation was determined by means of a capillary mercury dilatometer The power of the light beam that reached the unpolymerized material was regulated at 350 mW/cm2 (Group I) or 275 mW/cm2 (Group II) positioning the end of the light source (XL-3000, 3M-ESPE) either in contact with or 5 mm from the dilatometer stopper. Four measurements were taken for each material and experimental condition, reading the dilatometer scale one minute after light activation. Data were recorded as percentage volumetric shrinkage as a function of the material volume calculated from the specimen mass and density. Analysis of variance and Tukey's test were used for statistical evaluation. Shrinkage was found to be significantly greater in Group I - 1.43% (0.43) - than in Group II -1.04 % (0.38). Some significant differences were found among some of the products. A reduction in polymerization shrinkage occurred concomitantly with the reduction in light activation energy and varied according to the organic content and the physical and chemical characteristics of each material. PMID- 17121193 TI - Changes in blood pressure in children undergoing psychological treatment before dental procedures. AB - Different dental procedures in children undergoing treatment can induce changes in blood pressure. These changes will be minimized if psychological treatment is applied prior to any procedure. The objective of this study was to determine changes in blood pressure (systolic-diastolic) in children undergoing psychological treatment before dental procedures. The population studied consisted of 100 children and adolescents, ages 6 to 15 years. The patients were assigned to two groups at random, i.e. with or without psychological treatment before dental procedures. Behavior management was carried out applying the tell show-do technique of conduct handling. The systolic and diastolic blood pressures were measured prior to the initiation of the treatment, during drilling, restoring, and before dismissing the patient. The no-psychological treatment group exhibited significant differences in diastolic and systolic blood pressures during the dental procedure whereas the psychological treatment group evidenced no significant differences in blood pressure. No correlation was found between the alterations of blood pressure and the type of dental treatment performed. The application of the tell-show-do method would have an effect on the blood pressure in patients undergoing dental procedures. PMID- 17121194 TI - Early phenotypic and genotypic alterations in submandibular gland oncogenesis in rats. AB - The present study evaluates the phenotypic and genotypic changes that take place during early oncogenesis. The submandibular glands of male rats were injected with a 0.5% solution of 9,10-dimethyl-1,2-benzanthracene (DMBA) in acetone. Gland samples were taken at 0, 7, 30 and 150 days post-injection and submitted to histological, biochemical, immunocytochemical and PCR evaluation. Histopathological analysis was performed on hematoxylin-eosin stained slides. Total protein content was assessed by Lowry's method and the protein profile was analyzed by 12% SDS-PAGE. Bcl-2 was demonstrated by silver-enhanced gold immunolabeling. p53 immunolabeling was performed using the streptavidin-biotin system. All the treated animals developed carcinoma-like lesions at 30 and 150 days. Total protein concentration rose significantly (p < 0.05) above control values at 7, 30 and 150 days. The treated glands exhibited positive immunolabeling for p53 in the nuclei of neoplastic cells at 30 and 150 days. Treated glands also showed positive cytoplasmic immunolabeling for Bcl-2, exhibiting statistically significant differences between 7, 30 and 150 days (p = 0.0015), and with controls (p < 0.0001). No p53 mutations were observed whereas a point mutation, C-to-A, of the Bcl-2 gene was detected at 7, 30 and 150 days by PCR amplification. This mutation led to a single aminoacid change (thre --> asn) in the protein molecule. Our results suggest that the early histopathological changes correspond to quantitative and qualitative protein changes. The histopathological, biochemical, immunocytochemical and genetic alterations observed during the course of experimental carcinogenesis in the submandibular gland of the rat could constitute reproducible indices of malignant transformation applicable to human oncogenesis, given the high degree of homology between the oncogenes of mice, rats and human beings. PMID- 17121195 TI - Influence of short-term diabetes on osteocytic lacunae of alveolar bone. A histomorphometric study. AB - The aim of this study was to evaluate, for the first time, the histomorphometry of the cellular and lacunar features of the osteocytes of alveolar bone in acute streptozotocin-induced diabetic insulin-treated or untreated rats. Eighteen male Wistar rats weighing 200 to 260 g were assigned to one of the following groups: I) control group (C), II) diabetic group (DBT), and III) insulin treated diabetic group (DBT+INS). Experimental diabetes was induced by a single intraperitoneal injection of 60 mg/kg of body weight of streptozotocin. Insulin treatment began 24 h after the streptozotocin injection in animals of group DBT+INS in a dose of 4-6 IU of Humulin NPH insulin given as a single daily subcutaneous (s.c.) injection each morning between 07.00 and 10.00 h. The animals were euthanized on the 8th day. The upper maxillae were removed and fixed in buffered formalin, decalcified in EDTA, embedded in paraffin and stained with H-E for histologic and histomorphometric evaluation. Bone activity and lacunar density, osteocyte and empty lacunar densities, lacunar volume and lacunar shape were evaluated. Differences between variables were assessed by one-way ANOVA. Surface bone activity values revealed that bone resorption was significantly greater in the DBT group than in the C group (p < 0.05). Total lacunar density was significantly reduced in the DBT and DBT+INS groups as compared to control (p < 0.05). Concomitantly, a statistically significant reduction in osteocyte density and an increase, albeit not statistically significant, in empty lacunar density was observed in DBT and DBT+INS groups versus control. Lacunar volume did not exhibit statistically significant differences. The osteocyte lacunae in the DBT group lost their rounded shape and acquired intermediate shapes. This study reveals an early response of osteocytes to hyperglycemia, before systemic compensatory mechanisms are turned on. The effects are not always compensated by insulin treatment. PMID- 17121196 TI - Morphology of the lateral pterygoid muscle associated to the mandibular condyle in the human prenatal stage. AB - The lateral pterygoid muscle (LPM) inserts at the condyle and the articular disc and plays a central role in mandibular movement via the Temporomandibular Articular Complex. The aim of this study was to examine the association between the morphology of LPM muscular fascicles and the degree of mineralization of the mandibular condyle in the prenatal stage employing structural, ultrastructural and microanalytical evaluation. Sixteen human fetuses at 11-37 weeks of gestation, with no apparent pathology and resulting from spontaneous abortions, were included in the study. Samples from lateral pterygoid muscle and the mandibular condyle were processed for light microscopy and electron microscopy and microanalysis. Desmin immunolabeling (dilution 1: 25 Dako) and alpha sarcomeric actin immunolabeling (dilution 1:50 Dako) employing the avidin-biotin system were used in paraffin embedded samples. Contralateral samples were examine by transmission electron microscopy. Four condyles (at 17-21 weeks of gestation) were used to measure the relative content of calcium and phosphorous employing the X-ray diffraction microanalytical technique. At 11-16 weeks of gestation, the LPM was composed of secondary myotubes associated to satellite cells and nerve fibers. At 18 weeks, the muscle exhibited multiple compact fascicles and the condyle showed a thin, external, subperiostal mineralized layer with few central bone spicules. At 20 weeks, at the site of insertion of the LPM, the bone trabeculae of the condyle contained an electrondense matrix with abundant mineralization nuclei. At 17-21 weeks of gestation no significant variations in the contents of phosphorous and calcium were observed. At 24 weeks, transmission electron calcium and microscopy studies revealed a marked increase in the functional units of the muscle fascicles. Also, at this age muscle fibers exhibited differences in the expression of desmin and alpha sarcomeric actin. At 37 weeks the muscle became multipennate in appearance, exhibiting a more complex organization than younger fetuses. Alpha sarcomeric actin labeling became light with age. This results suggest that between 16 and 22 weeks of gestation the differentiation and maturation process of the muscle fibers precedes and prevails over the development and mineralization process from mandibular condyle. The rudimentary performance of the prenatal LPM would be one of the factors that regulate the process of ossification at the level of the mandibular condyle. The rate of ossification would increase starting from 22 of gestation week. PMID- 17121197 TI - In vivo bracket bond strength using two adhesive systems applied under wet and dry conditions. AB - The purpose of this study was to investigate, in vivo, the bond strength of two adhesive materials: a moisture insensitive primer (MIP)* and a one step self etching primer (SEP)*, both used with Transbond XT* on dry and wet enamel and an adhesion time of 10-15 minutes. First or second upper and/or lower bicuspids (n = 124), to be extracted for orthodontic reasons, were used. A comparison of the materials' behavior was conducted under four different situations: 1) MIP on enamel etched and dry; 2) MIP on a surface etched and wetted with patient's saliva; 3) SEP on a dry field, 4) SEP on a saliva-wet enamel. For statistical analysis, Dunn-Sidak's multiple comparison test was applied with a probability of less than 0.05 (before correction). Stainless steel brackets with mesh-backed pads were bonded to the teeth. Bond strength was tested with modified orthodontic pliers on which a strain-gage was fixed to measure handle deformation while debonding. Moisture insensitive primer tested on wet enamel showed the highest mean bond strength outcomes (8.98 MPa) compared to one step etching primer (5.81 MPa). Statistical difference between these groups was significant (p = 0.000). Standard deviation was lower for the one-step technique, under dry and wet conditions. Since the media bond strength of SEP proved sufficient for clinical purposes and its behavior tended to be more homogeneous, this was considered the best choice. PMID- 17121198 TI - Fine-needle aspiration of a Xp11.2 translocation/TFE3 fusion renal cell carcinoma metastatic to the lung: report of a case and review of the literature. AB - A 57-yr-old woman presented to the National Cancer Institute (NCI) with a history of nephrectomy for a clear cell renal cell carcinoma (RCC), Fuhrman grade 3 of 4 diagnosed 1 yr prior to admission to the NCI. A CT scan done upon admission revealed multiple bilateral lung masses. A CT-guided fine-needle aspiration (FNA) of one of the lung masses revealed a cellular specimen composed primarily of follicular structures surrounding dense hyalinized central cores. The cells in the follicular structures displayed bland nuclei and had granular to vacuolated cytoplasm. Papillary structures were also appreciated. Immunocytochemical studies showed tumor cells that were strongly vimentin and TFE3 positive. Focal staining for AE1/AE3 and CD10 was observed, as was negative staining for EMA. A surgical biopsy specimen reflected the FNA findings and demonstrated a similar immunoprofile. These findings correspond to the recently described Xp11.2 translocation/TFE3 fusion renal cell carcinoma. To our knowledge, this is the first report describing the cytologic features of an Xp11.2 translocation/TFE3 fusion RCC. PMID- 17121199 TI - Renal cell carcinoma with an Xp11.2 translocation in a 16-year-old girl: a case report with cytological features. AB - We report a case of a 16-year-old girl with a left renal tumor discovered by her family practitioner. On physical examination the patient had a painless abdominal mass, located in the upper medium portion of the abdomen on the left side with a voussure of the abdominal wall. Ultrasound and abdominal pelvic CAT scan revealed a large heterogeneous mass with calcifications in the inferior portion of the left kidney. We made touch-imprint cytological preparations of the biopsy fragments, obtained under ultrasound guidance. Cytological analysis revealed highly cellular smears with malignant cells arranged in large clusters or rarely isolated, sometimes surrounding hyaline nodules with numerous psammoma bodies. After May-Grunwald-Giemsa staining, cells displayed moderately irregular nuclei with an abundant and pale basophilic cytoplasm with well-defined borders and a finely granular texture. The diagnosis of a special type of renal cell carcinoma was suspected, and was then confirmed after examination of the biopsy sample and the corresponding surgical specimen. The histomorphologic features were those of a renal cell carcinoma associated with an Xp11.2 translocation. Immunohistochemistry revealed this translocation by showing nuclear positivity in tumor cells for an antibody raised against the TFE3 protein. The clinical outcome was marked several months later by metastases in lymph nodes, bone, lung, and adrenal gland as well as a local recurrence. PMID- 17121200 TI - Diagnostic pitfalls associated with fine-needle aspiration biopsy in a patient with the myxoid variant of monophasic fibrous synovial sarcoma. AB - Synovial sarcoma (SS) is one of the most common soft tissue tumors that typically presents in the extremities of young adults, but may occur at any site and affect children during the first decade. Herein we discuss a 12-yr-old male who complained of left foot pain and plantar mass. A fine-needle aspiration biopsy of an 8 cm subcutaneous mass was performed revealing a myxoid spindle cell neoplasm. The cytologic differential diagnosis included a myxoid neurofibroma, neurothekeoma, and a myxoid sarcoma. Subsequent excision of the mass revealed a monophasic fibrous SS with myxoid features. Examination of the tissue by fluorescence in situ hybridization confirmed the presence of characteristic SS SYT gene rearrangement at chromosome 18q11.2. This case underscores that the cytologic distinction of mxyoid spindle cell tumors may be challenging. We report the cytologic features of a myxoid monophasic fibrous SS, and discuss its distinction from other benign and malignant myxoid soft tissue neoplasms. PMID- 17121201 TI - Preventing cervical cancer. PMID- 17121202 TI - Disseminated nocardiosis diagnosed by fine needle aspiration biopsy: quick and accurate diagnostic approach. AB - Nocardia is an uncommon pathogen in immunocompetent patients; however, it has been increasingly recognized as a significant opportunistic pathogen in organ transplant patients. Diagnosis of Nocardiosis is usually made by microbiologic culture or cytologic examination of pulmonary specimens including, sputum, and brushing/washings or by histologic evaluation of tissue biopsy material. We report a case of subcutaneous Nocardiosis diagnosed by Fine-needle aspiration biopsy (FNA). The patient is a 66-year-old man with a history of lung transplantation and posttransplant lymphoproliferative disorder who presented with subcutaneous masses in the right upper arm and the left shoulder. FNA was performed in an outpatient clinic setting, with immediate morphologic assessment revealing filamentous branching organisms suspicious for Nocardiosis. Subsequent examination with special stains and microbiologic culture confirmed the diagnosis. The quick and accurate diagnosis by FNA led to emergent and appropriate treatment. PMID- 17121203 TI - State cancer survivorship policies. PMID- 17121204 TI - Metaplastic breast carcinoma with chondrosarcomatous differentiation: fine-needle aspiration cytology findings. A case report. AB - Metaplastic carcinoma (carcinoma with pseudosarcomatous metaplasia) of the breast are high-grade carcinomas in which much of the tumor undergoes metaplastic change producing a pseudosarcomatous pattern. We report a case of metaplastic breast carcinoma (MBC) in whom fine-needle aspiration (FNA) cytology was performed with later histological confirmation. The lesion affected a 68-yr-old woman, with a tumor measuring 6.4 x 5.3 cm well demarcated mass located in the upper outer quadrant of the right breast. FNA cytology revealed a variety of markedly atypical cells, mainly spindle-shaped, and mitotic figures sporadically distributed against a severely necrotic background. Atypical chondrocytes were observed against a background of myxomatous substance that displayed metachromasia with May-Giemsa stain that resembled chondrosarcoma cells. Clusters of markedly atypical carcinoma cells that exhibited epithelial junctions were also seen, and immunostaining confirmed the presence of both mesenchyme-marker positive sarcomatous and epithelial-marker-positive carcinoma cells. Careful attention to the precise cellular composition such as sarcomatous cells, chondrosarcomatous cells and carcinoma cells should allow the recognition of these neoplasms. Therefore, MBC seems to be very a characteristic tumor in which accurate cellular diagnosis may be achieved by FNA cytology. PMID- 17121206 TI - Sight test and glasses could dramatically improve the lives of 150 million people with poor vision. PMID- 17121205 TI - School wellness policies. PMID- 17121207 TI - WHO and Sanofi-Aventis expand programme to fight neglected tropical diseases. PMID- 17121208 TI - The World Health Organization calls for improved treatment for mental illness to reduce suicide. PMID- 17121209 TI - Global polio eradication now hinges on four countries. PMID- 17121210 TI - Insulin detemir: new drug. A second long-acting insulin analogue: many uncertainties, few advantages. AB - (1) The standard treatment for type 1 diabetes is intensive insulin therapy, consisting of at least 3 daily injections of different insulins, one of which is a long-acting insulin. (2) Insulin detemir is the second long-acting human insulin analogue to be marketed in Europe (after insulin glargine) for the treatment of diabetes in adults and children over 6 years of age. Its action lasts about 12 hours. (3) Insulin detemir was evaluated in around 10 comparative trials, all unblinded, examining the effect of insulin detemir in terms of global glycaemic control (HbA1c level). None of these trials examined whether insulin detemir prevented complications of diabetes. (4) About 10 trials, involving more than 3000 patients, showed that insulin detemir, insulin isophane and insulin glargine have similar efficacy in treating both type 1 and type 2 diabetes. (5) The short-term adverse effect profile of insulin detemir is similar to that of isophane insulin. There is slightly less weight gain with insulin detemir, but injection site reactions occur more frequently. The long-term adverse effects of insulin detemir are not known. (6) Insulin detemir is a clear solution, leading to a risk of confusion with ordinary human insulin or a fast-acting insulin analogue. (7) In practice, isophane insulin remains the first choice long-acting insulin for patients with type 1 or type 2 diabetes. PMID- 17121211 TI - Duloxetine: new indication. Depression and diabetic neuropathy: too many adverse effects. AB - (1) Several classes of antidepressants are available. The main difference between these classes is in their short-term pharmacological effects, leading to different patterns of adverse effects. Some antidepressants, especially tricyclics, have positive risk-benefit balances in the treatment of diabetic neuropathy. (2) Duloxetine, a compound chemically related to fluoxetine, appears to have a short-term mechanism of action similar to that of venlafaxine. In the European Union, duloxetine was first approved for female urinary stress incontinence. Another brand of duloxetine has since been marketed for depression and neuropathic pain in diabetic patients. (3) Duloxetine at a dose of 60 mg once a day showed moderate efficacy in 2 placebo-controlled trials. At this dose, however, there are no other comparative trials. It is therefore not possible to know whether duloxetine is as effective as other antidepressants. (4) Two placebo controlled trials involving patients with pain due to diabetic neuropathy concluded that a dose of 60 mg/day had efficacy, although of doubtful clinical relevance. In the absence of comparative trials, however, we do not know if this efficacy is even equivalent to that of a tricyclic antidepressant used as an analgesic. (5) In fibromyalgia, a controversial clinical diagnosis, two double blind placebo-controlled trials involving 207 and 354 patients failed to prove that duloxetine had tangible analgesic efficacy. It is therefore appropriate that this use is not mentioned in the "Indications" section of the summary of product characteristics (SPC). (6) The assessment of duloxetine in depression and neuropathic pain confirms existing data on its gastrointestinal, neuropsychological and hepatic adverse effects. In these trials, duloxetine increased blood pressure in a dose-dependent manner. (7) Duloxetine is metabolized by cytochrome P450 isoenzymes CYP 1A2 and CYP 2D6, creating an important risk of interactions with other drugs. (8) In practice, duloxetine currently has no place in the treatment of depression or diabetic neuropathy. Its efficacy has not yet been demonstrated to be even equivalent to that of other available drugs, and it has too many adverse effects, given this degree of uncertainty. PMID- 17121212 TI - What happens to the public interest when medical care becomes a business? PMID- 17121213 TI - Two cases of tetanus? AB - Tetanus is a rare infectious disease in Western countries that leads to diagnostic difficulties. Several diseases may mimic tetanus, and diagnostic considerations can at times be difficult, especially in critically ill patients, who need prompt treatment. Two patients are presented who were diagnosed with and treated for tetanus. However, the course of these patients' diseases strongly suggested that the diagnoses were incorrect. The article includes reflections on these 2 difficult diagnostic cases. These 2 cases illustrate the importance of proposing alternative diagnoses in suspected tetanus in Western countries. This may prevent loss of valuable time and prevent a delay of possibly effective therapy. PMID- 17121214 TI - [Twenty-five years of HIV and AIDS: opportunities and challenges]. AB - From the first report of AIDS case in the United States in 1981, to the development of effective antiretroviral drugs, researches on HIV/AIDS in the past 25 years made dramatic achievement. Breakthroughs were also made in the epidemiology, virology, immunology, and pathogenesis of HIV/AIDS, and in the development of blood tests for HIV. HIV/AIDS-related researches also made great advances in China in recent years. PMID- 17121215 TI - [Clinical and basic researches on HIV/AIDS]. AB - Worldwide, about 85% of human immunodeficiency virus (HIV) infections were acquired through sexual transmission. Control strategies have been focused on behavioral change through educational efforts and condom promotion, which had achieved certain success in several countries. The past decade witnessed the extraordinary advances in highly active antiretroviral therapy (HAART) and its effectiveness. HAART fundamentally alters the course of HIV-1 infection by decreasing the plasma viral load to the undetectable level and increasing the number of CD4 + T cells. However, problems including drug resistance and adverse events also exist in HAART. In the near future, the major challenges may include: determining the role and efficacy of new drugs and new therapies; addressing the coinfection of HIV/liver diseases/tuberculosis; improving the savage management; addressing the issues faced by the resource-limited countries; and achieving further success in HIV/AIDS prevention and treatment. PMID- 17121216 TI - [Resistance to HIV-1 infection of CD4 + T lymphocytes in vitro from chinese HIV-1 exposed seronegative individuals]. AB - OBJECTIVE: To determine the relative resistance to HIV-1 infection of CD4 + T lymphocytes in HIV-exposed seronegative individuals (ESNs) in China. METHODS: HIV primary isolates were obtained from peripheral whole blood of HIV-infected persons. CD4 + T lymphocytes of Chinese ESNs were separated from peripheral blood mononuclear cells with magnetic cell sorting (MACS). The purified CD4 + T lymphocytes were cocultured with HIV primary isolates. The p24 level was detected and the culture medium was refreshed every 3 days within 2 weeks. RESULTS: For M tropic HIV strains, p24 level was significantly lower in ESN group than in control group (P < 0.05); for some M tropic HIV strains, even no p24 replicated in ESN group. However, T tropic virus strains had no significant difference between these two groups (P > 0.05). CONCLUSION: CD4 + T lymphocytes of Chinese ESNs may possess relative resistance to M tropic HIV strains, which may be one of the main influencing factors that result in ESN. PMID- 17121217 TI - [Abnormal changes of CD28 expression on CD4 + T cells in treatment-naive and highly active antiretroviral therapy-treated HIV/AIDS patients]. AB - OBJECTIVE: To study the alteration of the expression of CD28 on CD4 + T cells in HIV/AIDS patients and observe the dynamics of CD28 expression under highly active antiretroviral therapy (HAART). METHODS: The expression of CD28 on CD4 + T cells, CD4 counts, and plasma viral load were measured by flow cytometry and bDNA assays in 278 treatment-naive HIV/AIDS patients and 56 healthy controls. In addition, the evolution of these parameters was assessed in 59 patients who initiated HAART and were followed for 12 months in regular 3-month visits. RESULTS: The median level of CD28 on CD4 + T cells decreased dramatically in treatment-naive HIV positive individuals than in HIV-negative controls (P <0.001). The expression rate of CD28 molecule was positively correlated with CD4 counts (r = 0.484, P < 0.001), and negatively correlated with plasma viral load (r = -0.300, P <0.001). In patients who had received one month of standard HAART, the level of CD28 on CD4 + T cells increased rapidly from 75.0% to 90.0% (P < 0.001). Moreover, there was a negative correlation between the median CD28 expression and the median viral load (r = - 0.829, P = 0.042). CONCLUSIONS: The level of CD28 expression on CD4 + T cells is down-regulated in treatment-naive HIV/AIDS patients. HAART can successfully restore the lymphocyte subsets of CD4 + CD28 + T cells. The up regulation of CD28 expression after HAART may be closely correlated with the suppression of the viral replication. PMID- 17121218 TI - [Pathology of AIDS-related lymphadenopathy and its relationship to expression of CD4 + CD25 + regulatory T cells in lymphoid node tissue]. AB - OBJECTIVE: To observe the pathology of AIDS-related lymphadenopathy and its relationship to the expression and distribution of CD4 + CD25 + regulatory T cells in lymphoid node tissue. METHODS: Totally 22 biopsy and 13 autopsy lymphoid node tissues from HIV-positive patients were examined under microscopy and pathological staging was performed. Specific marker for CD4 + CD25 + regulatory T cells in lymphoid node tissue was detected with anti-Foxp3 monoclonal antibody by immunohistochemistry. RESULTS: Among all the 35 specimens, 5, 4, 14, and 12 specimens were histopathologically staged from 1 to 4, respectively. FoxP3 were detected in all lymphoid node tissues. The distribution of FoxP3-positive lymphocytes were mainly in intermediate zone of follicle and cortical area in stages 1 and 2. The counts of FoxP3-positive lymphocytes remarkably decreased in stages 3 and 4, following depletion of lymphocytes. CONCLUSIONS: CD4 + CD25 + regulatory T cells exist in lymphoid node tissue of patients with HIV infection. Their amounts decrease or deplete along with the progression of AIDS-related lymphadenopathy. PMID- 17121219 TI - [Inhibition of expression of CCR5 and CXCR4 on cells by adenovirus-mediated antisense RNA]. AB - OBJECTIVE: To suppress the expression of CCR5 and CXCR4, the co-receptors for human immunodeficiency virus type 1 ( HIV-1), and thus inhibit HIV-1 from entering cells. METHODS: DNA fragments encoding either CCR5 or CXCR4 were amplified from healthy human peripheral blood mononuclear cells (PBMCs) by reverse transcript polymerase chain reaction (RT-PCR) and sequencing was performed. Correct fragments were inserted into Shuttle plasmid inversely, which was recombined with backbone plasmid containing homologous adenoviral genome in E. coli BJ5183. The recombinant plasmids were transfected into 293 cells in which they were packaged and amplified. Recombinant adenoviruses containing antisense RNA of CCR5 or CXCR4 were obtained and identified by RT-PCR, and the titres of them were determined by cytopathic effect (CPE) method. The U937 and MT4 cells were infected by recombinant adenoviruses containing antisense RNA of CCR5 (multiplicity of infection, MOI = 100) and CXCR4 (MOI = 200), respectively. The expression of co-receptors on infected cell was measured by fluorescence activated cell sorter at 24, 48, 72 hours and 10 days after infection. In addition, the chemotactic activity and proliferation of infected cells were detected with Boyden chamber and 3H incorporation respectively. RESULTS: We constructed the recombinant plasmids and obtained the recombinant adenoviruses which contained antisense RNA of CCR5 or CXCR4 and were designated as pAd-antiR5 and pAd-antiX4 respectively. The titers of recombinant adenoviruses pAd-antiR5 and pAd-antiX4 were 5 x 10" PFU/ml and 7 x 10(10) PFU/ml, respectively. The expression rate of CCR5 on U937 cells decreased from 82. 10% (blank control) to 1.12% (Ad-antiR5 infected) , and that of CXCR4 on MT4 cells decreased from 42% (blank control) to 1.03% (Ad-antiX4 infected) 24 hours later. The expression rates of CCR5 on Ad-antiR5 infected U937 cells were 1.02% , 1.26% , 1.23% at 48 hours, 72 hours, and 10 days later, respectively. The expression rates of CXCR4 on Ad-antiX4 infected MT4 cells were 1.13%, 1.17%, 1.22% at 48 hours, 72 hours, and 10 days later, respectively. Moreover, the recombinant adenovirus had no effects on chemotactic activity and proliferation of the cells. CONCLUSION: The recombinant adenovirus containing antisense CCR5 or CXCR4 can remarkably decrease the expression of co-receptors for HIV-1 on U937 or MT4 cells without affecting their chemotactic activities and proliferative abilities. PMID- 17121220 TI - [Background study of HIV-1 drug resistant mutations in treatment-naive patients in liaoning province]. AB - OBJECTIVE: To collect background information on drug resistance mutations in treatment-naive HIV-1 infected individuals in Liaoning Province. METHODS: Samples from 91 antiretroviral therapy-naive patients were collected. The entire protease gene and 1-290 amino acids of the reverse transcriptase gene were amplified by nested PCR from provirus DNA and sequenced. The results were analyzed with HIVdb Drug Resistance Algorithm, and genotypic resistance mutations were determined to particular anti-HIV drugs. RESULTS: Totally 91 sequences were obtained, 3 of which displayed M46I mutations in the protease gene. Minor resistance mutation rate to protease inhibitors was 100%, including types of L63P (60.4%), V77I (60.4%), M36I/V (31.9%), A71V/T (22.0%), L10I (8.8%), and K20R (6.6%). Only one sequence carried reverse transcriptase related resistance mutations M184I. CONCLUSIONS: About 4.4% of HIV-1 infected individuals in Liaoning Province carried strains with drug resistance mutations. Most treatment-naive HIV-1 infected individuals in Liaoning Province were sensitive to the currently available antiviral medicines, but antiviral treatment must be in accordance with the strict procedures to keep better adherence and avoid the prevalence of drug resistant strains. PMID- 17121221 TI - [Human immunodeficiency virus type 1 strains epidemic in Shenzhen]. AB - OBJECTIVE: To study the epidemic status of human immunodeficiency virus type 1 (HIV-1) subtypes in Shenzhen and to study their transmission source and routes. METHODS: HIV-1 env and gag genes were amplified by nested PCR from uncultured peripheral blood mononuclear cells (PBMCs) obtained from 122 HIV-1 carriers confirmed in Shenzhen. The C2-V3 region (about 450 bp) of HIV-1 env and P17/ P24 region were sequenced. RESULTS: Among 122 samples, 6 HIV-1 strains including 3 circulating recombinant forms (CRFs) of CRF01_AE, CRF08_BC, CRF07_BC and 3 subtypes of B', B, C were found in Shenzhen, and the percentages were 45.1% (55/122) for CRF01_AE, 31.1% (38/122) for CRF08_BC, 6.6% (8/122) for CRF07_BC, 14.8% (18/122) for B' subtype, 1.6% (2/122) for B subtype, and 0.8% (1/122) for C subtype. The intragroup genetic distances were (4.455 +/- 1.478)%, (2.997 +/- 1.345)%, (4.380 +/- 2.024)%, (5.186 +/- 2.487)%, and (4.869 +/- 2.638)%, respectively. In comparison with the sequence of respective international strains 01AE. TH. 90. CM240, 97CNGX-9F, CN. 97. C54A, B. US. 83. JRFL, and RLA2, the genetic distances were (5. 228 +/- 0.823)%, (3.634 +/- 1.073)%, (4.233 +/- 1.119)%, (4.950 +/- 2.564)%, and (5.795 +/- 2.198)%, respectively. The major subtypes found in injection drug users (IDUs) were CRF07_BC, CRF08_BC, and CRF01_AE strains. CRF01_AE and B' strains were epidemic mainly in sexual workers. CONCLUSION: There are 3 HIV-1 subtypes (B', B, C) and 3 CRFs (CRF01_AE, CRF08_BC, CRF07_BC) epidemics in Shenzhen. The predominant subtypes varies among different transmission routes. While CRF01_AE is predominant among sexual workers, CRF08_BC and CRF01_AE are major subtypes among IDU population. PMID- 17121222 TI - [Expression of glutathione-S-transferase fusion protein and human CCL3L1 protein]. AB - OBJECTIVE: To clone human CCL3L1 cDNA and to express and purify the glutathione-S transferase (GST) fusion protein and human CCL3L1 protein. METHODS: Total RNA was isolated from breast cancer cell line MCF7. CCL3L1 cDNA including open reading frame was obtained by RT-PCR. PCR product was digested with EcoR I and cloned into the pGEX-4T-1 vector. The plasmids from positive clone was prepared and sequenced to confirm the CCL3L1 in correct fusion form. pGEX-4T-CCL3L1 was transfected to BL21 E. coli via isopropyl-beta-D-thiogalactoside (IPTG) induction to produce GST-CCL3L1 fusion protein, which was further detected by SDS-PAGE and Western blotting. RESULTS: As shown and confirmed by restriction endonuclease digestion analysis, CCL3L1 was correctly inserted into pGEX-4T-1 vector. The expressed fusion protein had a relative molecular weight of approximately 34 kD. CONCLUSION: GST-CCL3L1 fusion protein can be successfully expressed using appropriate vector. PMID- 17121223 TI - [Comparison of immunological profiles between pediatric and adult patients with AIDS in China]. AB - OBJECTIVE: To compare the immunological profiles of pediatric and adult patients with AIDS in China. METHODS: Totally 103 pediatric AIDS patients, 38 adult patients, 88 healthy children, and 72 healthy adults were enrolled. CD4 + T lymphocyte counts were determined by four-color flow cytometer and HIV-RNA levels were measured in EDTA plasma by quantitative reverse-transcription polymerase chain reaction (RT-PCR). Plasma levels of interleukin (IL)-10, IL-16, IL-18, regulated upon activation, normal T-cell expressed and secreted (RANTES), monocyte chemoattractant protein 1 (MCP-1), stromal cell-derived factor-(SDF-1) alpha, SDF-1 beta, and macrophage stimulate protein (MSP) were quantified by enzyme-linked immunosorbent assay (ELISA). The levels of beta 2-microglobulin (beta 2-MG) and soluble Fas (sFas) were measured to indicate the activation of immune system. RESULTS: The mean CD4 + T cell count in pediatric patients with AIDS was significantly lower than in healthy children (P < 0.01), as between the adult AIDS patients and healthy adults (P < 0.01). The mean levels of these cytokines in pediatric patients were significantly higher than in healthy children (P < 0.01). The level of MSP in adult patients was significantly lower than in healthy adults and other cytokines were significantly higher (P < 0.01). The mean levels of these cytokines, except SDF1 alpha and beta 2-MG, were significantly higher in pediatric patients than in adult patients (P < 0.01). CONCLUSION: Abnormal immune activation is induced in both pediatric and adult patients with HIV-1 infection. The level of immune activation is higher in pediatric patients than in adult patients. PMID- 17121224 TI - [Clinical characteristics of 143 Chinese HIV/AIDS patients]. AB - OBJECTIVE: To investigate the clinical characteristics of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients in China. METHODS: Totally 143 HIV/AIDS patients who were first diagnosed in Peking Union Medical College Hospital form January 1988 to April 2006 were enrolled in this study. Clinical characteristics were retrospectively analyzed. RESULTS: Among 143 HIV/ AIDS patients, 57 patients had no clinical symptoms and were confirmed by routine examinations; 86 patients had clinical symptoms, including fever (n = 50), weight loss (n = 18), and discomforts involving respiratory system (n = 34), gastrointestinal system (n = 16), and derma and mucosa (n = 17). Opportunistic infections (OIs) such as pneumocystis jiroveci pneumonia (PCP) (n = 27), oropharyngeal candidiasis (n = 16), tuberculosis (n = 15) , and cytomegalovirus (CMV) infection (n = 9) were also observed in patients whose CD4 + T cell counts were less than 200/mm3. Most CMV infection and cryptococcal meningitis occurred in patients whose CD4 + T cell counts were less than 100/mm3. CD4 + T cell count was negatively correlated with plasma viral load (r = -0.420, P = 0.001). CONCLUSIONS: Fever, dyspnea, and weight loss are the most common symptoms in the patients of this study. The respiratory system, gastrointestinal system, derma and mucosa are the most commonly affected areas by OIs, and PCP is the most common OI. The occurrence of OIs corelates with CD4 + T cell count. PMID- 17121225 TI - [Epidemiology of pediatric HIV infection in six provinces of China]. AB - OBJECTIVE: To investigate the epidemiology of pediatric human immunodeficiency virus (HIV) infection in six provinces of China. METHODS: A cross-sectional study was conducted in six provinces with the highest HIV prevalence. Surveys on demographics and HIV-related questions (transmission modes, time of diagnosis, clinical stage, laboratory test) were distributed to clinicians in these provinces. Descriptive and bivariate analyses were performed on the completed surveys. RESULTS: Survey results of 650 children [405 males and 245 females; average age: (7.9 +/- 3.2) years] were eligible for analysis. The interval between possible transmission and diagnosis was (7.1 +/- 3.2) years. The location distribution was as follows: 570 cases (87.7%) in Henan Province, 23 cases (3.5%) in Guangxi Province, 21 cases (3.2%) in Yunnan Province, 19 cases (2.9%) in Hubei Province, 10 cases (1.5%) in Anhui Province, and 7 cases (1.1%) in Shanxi Province. Transmission routes included mother-to-child transmission (75.1%), blood transfusion/ plasma donation (15.7 %), and injecting drug use (IDU, 0.5%). Former plasma donation (FPD) was the main transmission route in some provinces (Henan, Shanxi, Hubei, and Anhui), while IDU was the main transmission route in other provinces (Guangxi and Yunnan). The average age in the FPD provinces was significantly higher than that in IDU provinces [(8.1 +/- 3.2) vs. (5.4 +/- 2.2) years, P <0.001]. Among 178 patients in all six provinces who required treatment (on the basis of CD4 count or WHO staging), 133 (74.7%) did not receive treatment and 45 (25.3%) received antiretroviral therapy. CONCLUSION: Mother-to-child transmission is the main transmission mode in pediatric patients. Efforts should be made to strengthen the diagnosis and treatment of pediatric HIV/AIDS patients. PMID- 17121226 TI - [Effects of traditional Chinese medicine on CD4 + T cell counts and HIV viral loads during structured treatment interruption in highly active antiretroviral therapy]. AB - OBJECTIVE: To explore the impacts of traditional Chinese medicine (TCM) on CD4 + T cell counts and human immunodeficiency virus (HIV) viral loads during the course of structured treatment interruption (STI) in highly active antiretroviral therapy (HAART). METHODS: Nineteen HIV/ADIS patients were treated for 14 months as follows: initiated with zidovudine/lamivudine + efavirdine for 6 months, then discontinued the therapy and treated with TCM instead for 2 months. HAART was then reinitiated for another 3 months, and then discontinued and replaced with TCM for another 3 months. The changes of CD4 + T cell counts and HIV viral loads were measured. RESULTS: During the first STI of HAART, 43.8% of patients had no viral rebounds one month later, and 62.6% had stable or increased immune functions; 18.8% had no viral rebounds two months later, and 43.8% had stable or increased immune functions. Changes of viral loads were not significantly different between these two months (P = 0.097), while CD4 + T cell counts significantly decreased two months later compared with one month later (P = 0.043). During the second STI of HAART, 33.3% of patients had no viral rebounds one month later, and 64.3% had stable or increased immune functions; 13.3% had no viral rebounds 3 months later and 46.6% had stable or increased immune functions. Changes of viral loads had significant difference (P = 0. 017), while CD4 + T cell counts at month 12 elevated significantly compared with the baseline (P = 0.014). CONCLUSIONS: TCM can suppress the viral rebounds during STI-HAART, maintain immune functions. However, this effect may decrease along with the prolongation of STI-HAART. PMID- 17121227 TI - [Coinfection with HBV and HCV in 128 AIDS patients infected through blood transmission]. AB - OBJECTIVE: To analyze the clinical features of 128 acquired immunodeficiency syndrome (AIDS) patients infected through blood transmission who were coinfected with hepatitis B virus (HBV) and hepatitis C virus (HCV). METHODS: The prevalence, liver functions, and some immunological profiles of 128 AIDS patients coinfected with HBV and HCV were retrospectively analyzed. RESULTS: Among the 128 AIDS patients, 107 (83.6%) were coinfected with HCV, among which 40 (31.3%) patients had abnormal liver functions or liver damage and 15 (11.7%) patients experienced hepatitis symptoms. Three (2.3%) AIDS patients were singly coinfected with HBV, and all of them had abnormal liver functions and hepatitis symptoms. Seven (5.5%) patients were coinfected with HIV/HCV/HBV and none of them had abnormal liver functions or hepatitis symptoms. Eleven (8.6%) patients were only infected with HIV. CONCLUSIONS: The prevalence of blood-transmitted HIV patients coinfected with HCV is higher than with HBV. The clinical outcomes of HIV coinfection with HCV and HBV are different. PMID- 17121228 TI - [A novel anti-DNA antibody modified coronary stent for site-specific plasmid DNA delivery]. AB - OBJECTIVE: To explore the feasibility of using an endovascular metal stent as a highly efficient and site-specific gene delivery system. METHODS: Stents were formulated with a collagen coating. Anti-DNA monoclonal antibodies were covalently bound to the collagen surface by a cross linking reagent of N succinimidyl 3-(2-pyridyldithio) propionate (SPDP). Binding capacity and stability of antibody and plasmid DNA on stents were quantified by radioactive labeling. The gene transduction efficiency was evaluated in cell culture and in rabbits. RESULTS: The amount of antibodies binding on collagen matrix through SPDP reaction was 15 times higher than that of through physical absorption (P < 0.005). The binding stability of plasmid was significantly better than the control groups (P < 0.01). There was no harmful effect on cell growth with the anti-DNA antibody modified stents. The stents retrieved from cell culture after 72 hours of incubation in A10 cells showed numerous transducted cells only infiltrating the surface coating indicating a highly localized and efficient gene delivery pattern. Results of in vivo gene transfer by this modified stent revealed (2.8 +/- 0.7)% of total cells transduction and the higher transduction location was neointimal layer (about 7%). No distal spread of vector was detectable in the anti-DNA antibody modified stent implantation animals. CONCLUSIONS: Anti-DNA antibody modified stents represent a novel highly efficient and site-specific gene delivery system which can deliver various kinds of plasmid vectors. The release of plasmid DNA tethered on the stents could be controlled in some conditions. This novel system provided a novel platform for cardiovascular site-specific gene therapy. PMID- 17121229 TI - [Effect of silencing hypoxia-inducible factor-1 alpha by RNA interference on human breast carcinoma cell line]. AB - OBJECTIVE: To investigate the effect of short hairpin RNA (shRNA) targeting hypoxia-inducible factor-1 alpha (HIF-1 alpha) on the human breast carcinoma MCF 7 cell line. METHODS: The hypoxia environment was achieved by treating cells with cobalt chloride. The shRNA eukaryotic expression vector targeting HIF-1 alpha was constructed, and transfected into MCF-7 cells through lipofectamine 2000. Semi quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to study the expression of vascular endothelial growth factor (VEGF). The mRNA and protein level of HIF-1 alpha were detected by real-time PCR and Western blot. Sub G1 apoptotic population analysis, Annexin V/PI binding assay, and DNA ladder analysis were applied to investigate the cell apoptosis. The cell cycle was detected by flow cytometry. RESULTS: The mRNA and protein level of HIF-1 alpha increased after exposure of MCF-7 cells to hypoxia (P < 0.01). However, apoptosis was lower in hypoxia compared with normoxia (P < 0.05). The HIF-1 level of MCF-7 transfected with HIF-1 alpha shRNA decreased approximately 91.63% (P < 0.01). When the cells were treated with or without apoptosis inducer Ara-C, the apoptosis of MCF-7 cells transfected with HIF-1 alpha shRNA increased by 1.75 times (P < 0.01) and 61. 31 times (P < 0.01), respectively. The expression of VEGF in MCF-7 cells transfected with HIF-1 alpha shRNA decreased 66.8% compared with untransfected cells (P < 0.05). Cell cycle progression was inhibited when the MCF-7 cells were transfected with HIF-1 alpha shRNA. CONCLUSIONS: HIF-1 alpha plays an anti-apoptotic role in human breast carcinoma MCF-7 cell line. The shRNA we designed targeting HIF-1 alpha in MCF-7 can promote cell apoptosis, inhibit the expression of VEGF, and delay cell cycle progression. PMID- 17121230 TI - [Phenomena and pathological significances of the methylated p16 promotor in DNA derived from plasma and blood cells of patients with systemic lupus erythematosus]. AB - OBJECTIVE: To detect the methylation status of p16 gene promotor in DNA derived from plasma and blood cells of patients with systemic lupus erythematosus (SLE) , and it's relationship with clinical symptoms. METHODS: p16 promotor methylation in plasma and peripheral blood cells (PBCs) DNA were simultaneously detected with the methylation specific PCR (MSP) method in 24 active SLE patients, 21 inactive SLE patients, as well as 20 healthy controls. RESULTS: In the plasma DNA, p16 gene methylation ratio (MP%) was higher in SLE patients than in the healthy controls (64.4% vs. 5.0%, P < 0.05). MP% in the active SLE patients was significantly higher than that in the inactive SLE patients (83.3% vs. 42.9%, P < 0.05). In the PBCs, p16 gene methylation ratio (MC%) in the healthy controls was significantly higher than that in SLE (80.0% vs. 48.9%, P < 0.05). MC% in the active SLE patients (29.2%) was the lowest among three groups. There was no significant difference between the inactive SLE patients and healthy controls (71.4% vs. 80.0%, P > 0.05). Each patient could be judged as one of the four methylation patterns: MP/MC, UP/MC (UP: unmethylated plasma p16) , MP/UC (UC: unmethylated PBCs p16) , and UP/UC. The ratios of MP/ MC and UP/UC were similar between the active and inactive SLE patients. However, different distributions of other two patterns were found in the active and inactive SLE patients as UP/MC 4.2% vs. 42.9% (P <0.05) and MP/UC 58.3% vs. 14.3% (P < 0.05), respectively. The active SLE patients with MP/UC and the inactive SLE patients with UP/MC showed different clinical symptoms and laboratory examinations. Significant correlation was found between the disease activity index for lupus patients (SLEDAI) scores and MP% (r = 0.93), between the SLEDAI scores and MC% (r = - 0.96) also between MC% and MP% (r = - 0.79). CONCLUSION: The p16 methylation assay provides available information for the diagnosis, judgment of disease activity, as well as novel insights into the pathogenesis underlying this disease. PMID- 17121231 TI - [Animal study of intravascular gene therapy based on polyurethane implantable devices]. AB - OBJECTIVE: To explore the feasibility of utilizing two implantable devices made from modified polyurethane films with antibody tethered replication-defective adenoviruses encoding for green fluorescent protein (AdGFP) as gene delivery platforms. METHODS: Intra-aortic button implants of collagen-coated polyurethane films with antibody tethered AdGFP were sutured into the infrarenal aorta of adult pigs and pulmonary valve leaflet in juvenile sheep was replaced by polyurethane pulmonary valve cusp replacement with antibody-tethered AdGFP. After seven days, the buttons, prosthetic leaflets, and their surrounding tissues were explanted and evaluated for biocompatibility and AdGFP-mediated gene transfer by fluorescent microscopy and PCR analysis. RESULTS: In vivo analysis of gene transfer from collagen-coated polyurethane films in pig infrarenal aorta implants, one week explants of the collagen-coated polyurethane films demonstrated (14.2 +/- 2.5)% of neointimal cells on the surface of the implant. In sheep pulmonary valve leaflet replacement studies, polyurethane films with antibody tethered AdGFP vector demonstrated (25.1 +/- 5.7)% of cells attached to polyurethane valve leaflets were transduced in one week. PCR analyses showed that GFP DNA was not detectable in blood or distal tissues. CONCLUSION: Site-specific intravascular delivery of adenoviral vectors for gene therapy can be achieved with these two kinds of polyurethane implants utilizing the antivector antibody tethering mechanism. PMID- 17121232 TI - [Preparation of tetrandrine solid lipid nanoparticles]. AB - OBJECTIVE: To prepare solid lipid nanoparticles (SLN) loaded tetrandrine (TET) extracted from traditional Chinese medicine with ultrasonication and high pressure homogenization, and to compare the physicochemical characteristics of the particles produced by the two methods. METHODS: TET was incorporated into SLN by ultrasonication and high pressure homogenization separately. Transmission electron microscopy was employed to study the shape. Particle characterization system and Zeta potential analyzer were used to study the diameter and Zeta potential of SLN in suspension. The entrapment efficiency was determined with the high-performance liquid chromatography. The stability of SLN was also studied. RESULTS: TET-SLNs prepared by these two methods were sheet-shaped and irregular, but the SLNs prepared by high pressure homogenization were smaller. The mean diameter of SLN prepared by ultrasonication was (92 +/- 6) nm with Zeta potential of (-21.11 +/- 2.12) mV in distilled water, and the mean entrapment efficiency was 95.27%. The mean diameter of TET-SLN prepared by high pressure homogenization was (47 +/- 3) nm with Zeta potential of (-32.99 +/- 2.54) mV, and up to 97.82% of TET was incorporated. The diameter of SLN prepared by high pressure homogenization and ultrasonication were (52 +/- 5) nm and (168 +/- 12) nm after 90 days of storage at room temperature. CONCLUSION: Compared with ultrasonication, high pressure homogenization is a better method to prepare TET SLN, which is smaller, steadier and highly incorporated. PMID- 17121233 TI - [Treatment of hepatocellular carcinoma in mice with locally administered epirubicin-loaded poly(D, L)-lactic acid microspheres]. AB - OBJECTIVE: To study the effectiveness of treating hepatocellular carcinoma (HCC) in mice with locally administered epirubicin-loaded poly( D, L) - lactic acid microspheres (EPI-PLA-MS ). METHODS: EPI-PLA-MS was prepared with double emulsion solvent evaporation technique. Five groups of mice (n = 8 in each group) were intraperitoneally injected with five different doses of free epirubicin (FEPI), and the maximum tolerated dose (MTD) was calculated. Then 15 mice with transplanted subcutaneous H22 HCC were divided into three groups (n = 5), which were respectively intratumorally injected with normal saline (NS), blank microspheres, and EPI-PLA-MS (with 9 mg/kg of EPI). After two weeks the tumors were excised and weighed. Another 15 mice with transplanted H22 ascites HCC were divided into three groups (n = 5), which were intraperitonealy injected with the same drugs, and the increased life span were registered exactly. RESULTS: The MTD of intraperitoneally injected FEPI was 9 mg/kg. The tumour-inhibiting rates was 40.35% and 36.09% when EPI-PLA-MS were administered by intratumoral injection to the mice with subcutaneous H22 HCC. It significantly prolonged the survival time of mice with H22 ascites HCC and the increased life span by 153.49% and 142.22% when EPI-PLA-MS were intraperitoneally administered. CONCLUSION: EPI-PLA-MS is a new sustained-release preparation with high-efficacy and low-toxicity in treating HCC and has shown promising prospects when administered locally. PMID- 17121234 TI - [8B7 spectrin--a new member of spectrin family]. AB - OBJECTIVE: To analyze the nature of the protein encoded by 8B7cDNA (1 835 bp) and to examine the localization of the protein in cells. METHODS: The nature of the protein was analyzed using Blastn, Blastp, and TMpred. Expressions of 8B7 mRNA in tissues and cells were examined by Northern blotting. Recombinant expression vectors for localization test were constructed and transfected into COS-7 cells. Fluorescence emission in cells was examined upon a laser scan confocal microscope. RESULTS: The protein encoded by 8B7cDNA was 363 amino acids long and contained spectrin repeats. It was completely homologous to the C-terminal 363 amino acids of Enaptin, Nasprin-1, Mynel, and Syne-1 proteins. It belonged to Spectrin super-family and was called 8B7 spectrin. Northern blotting revealed a 1.8 kb mRNA expression in human spleen and small intestine tissues. EGFP-8B7 fusion protein was observed to express on the nuclear membrane and in the cytoplasm in a reticular form in a localization assay on COS-7 cells. The position of fluorescence in COS-7 cells transfected with pEGFP-delta SR8B7 was similar to that in the cells transfected with pEGFP-8B7cDNA. CONCLUSIONS: 8 B7 spectrin is a novel member of spectrin super-family. It localizes to the nuclear membrane and the cytoplasm in a reticular form in COS-7 cells. The localization is determined by the C-terminal KASH domain in 8B7 spectrin molecule. PMID- 17121235 TI - [Identification of traditional Chinese medicine scorpio by powder X-ray diffraction Fourier pattern]. AB - OBJECTIVE: To set up a new analysis method of the traditional Chinese medicine Scorpio. METHODS: Ten Scorpio samples were obtained from Hubei, Shaanxi, and Shandong provinces and analyzed with X-ray diffraction Fourier pattern technique to obtain the geometric topology and characteristic marked peak of Scorpio. RESULTS: The geometric topologies of 9 samples were similar, excepting Sample 7#. Totally 11 characteristic marked peaks were observed among these 9 samples. CONCLUSION: X-ray diffraction Fourier pattern is a useful tool for the identification and quality control of the Scorpio. PMID- 17121236 TI - [Association between genetic polymorphism in xeroderma pigmentosum G gene and risks of laryngeal and hypopharyngeal carcinomas]. AB - OBJECTIVE: To study the association between polymorphism of DNA repair gene xeroderma pigmentosum G (XPG) Asp1104His and the risks of developing laryngeal and hypopharyngeal carcinomas. METHODS: Totally 175 patients with laryngeal or hypopharyngeal carcinoma and 525 cancer-free controls were genotyped for the polymorphism by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The odds ratio (OR) and 95% confidence interval (CI) were calculated using unconditional logistic regression model. RESULTS: Compared with those having the Asp/Asp genotype, patients having the XPG 1104Asp/His genotype had a higher risk for laryngeal carcinoma (OR = 2.46, 95% CI = 1.15 5.24, P < 0.05), but not for hypopharyngeal carcinoma (OR = 1.36, 95% CI = 0.87 2.12, P > 0.05). In addition, the XPG 1104Asp/His genotype appeared to be associated with well differentiated squamous cell carcinoma in both larynx and hypopharynx (OR = 1.88, 95% CI = 1.05-3.40, P < 0.05 ). CONCLUSION: The XPG Asp1104His polymorphism may play a role in the development of laryngeal and hypopharyngeal carcinomas. PMID- 17121237 TI - [Evaluation of oxidative stress in experimental rabbit aneurysms]. AB - OBJECTIVE: To evaluate the roles of oxidative stress in the generation and development of aneurysms. METHODS: Five terminal aneurysms and 8 lateral aneurysms were rebuilt on rabbits, and 6 normal artery vessels were prepared as control. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), and anti-reactive oxygen species unit (anti-ROS unit) were measured with chemical methods. RESULTS: The levels of MDA in terminal and lateral aneurysms were (33.85 +/- 8.66) and (27.87 +/- 5.78) nmol/mg prot, respectively, which were significantly higher than (10.91 +/- 2.72) nmol/mg prot in control group (P < 0.01). The levels of SOD in terminal and lateral aneurysms were (28.30 +/- 3.58) and (33.00 +/- 8.09) U/mg prot, respectively, which were significantly lower than (127.27 +/- 38.72) U/mg prot in control group (P < 0.01). The levels of anti-ROS unit in terminal and lateral aneurysms were (47.86 +/- 5.00) and (62.64 +/- 13.87) U/ mg prot, respectively, which were significantly lower than (116.94 +/- 9.22) U/mg prot in control group (P < 0.01). No significant differences were shown between terminal aneurysm and lateral aneurysm in MDA and SOD except anti ROS unit (P = 0.014). MDA had negative correlations with both SOD and anti-ROS unit, and the correlation coefficients were -0. 830 and -0. 852, respectively. CONCLUSION: Oxidative stress may play an important role in the development of aneurysms. Oxidative stress seems similar among various aneurysms. PMID- 17121238 TI - [Preparation and characterization of a new injectable bone substitute carrageenan/nano-hydroxyapatite/collagen]. AB - OBJECTIVE: To develop a new injectable biomaterial carrageenan/nano hydroxyapatite/collagen (nHAC/Carr) for bone surgery and characterize it. METHODS: nHAC/Carr was developed by filling carrageenan with nano hydroxyapatite/collagen (nHAC) granules. RESULTS: It was found that nHAC/Carr had similar X-ray diffraction patterns with that of nHAC. Fourier transform infrared spectroscopy showed that carrageenan did not change its structure in nHAC/Carr. The rheological behavior of nHAC/Carr was the same as carrageenan. The scanning electron microscopy and porosity analysis showed that nHAC/Carr had porous structure and its porosity was about 90%. CONCLUSION: The biomaterial nHAC/Carr may be used as an injectable bone substitute. PMID- 17121239 TI - [Range of plasma hydrogen sulfide in children]. AB - OBJECTIVE: To measure the range of plasma hydrogen sulfide (H2S) in children. METHODS: Totally 200 healthy children were classified into 4 groups based on age and sex: 7-14 years old group (n = 75, 43 boys and 32 girls), 15-19 years old group (n = 125, 64 boys and 61 girls). Plasma H2S level was detected by a modified sulfide electrode-based method. RESULTS: Plasma H2 S levels were (52.2181 +/- 17.9400) micromol/L in 7-14 years old boys, (51.9441 +/- 16.5448) micromol/L in 7-14 years old girls, (52.8771 +/- 14.1444) micromol/L in 15-19 years old boys, and (53.6551 +/- 14.5563) micromol/L in 15-19 years old girls (P > 0.05). In summary, the range of plasma H2S in children was about (52.8234 +/- 15.4339) micromol/L. CONCLUSION: The range of plasma H2S in children is about (52.8234 +/- 15.4339) micromol/L. PMID- 17121240 TI - [Changes of matrix metalloproteinases activities in pulmonary fibrosis rats]. AB - OBJECTIVE: To observe the changes of matrix metalloproteinases (MMPs) activities in pulmonary fibrosis rats. METHODS: Eighty male SD rats were randomly divided into sham group (n = 40) and bleomycin group (BLM, n = 40), in which SD rats were injected with a single intratracheal dose of sham saline or bleomycin respectively. On day 1, 3, 7, 14, and 28 following bleomycin or saline instillation, rats were randomly killed, and serum from abdominal aorta, alveolar fluid from the bronchoalveolar lavage, and the lung homogenate were collected and then stored at -80 degrees C. MMPs activity was determined by zymography. RESULTS: Compared with sham group, the levels of MMP-9 in all samples were augmented. MMP-9 activities in the serum were highest on day 3 than those on day 1 and day 7, and in lung tissue homogenate were highest on day 7; however, no significant differences were found between BLM group and sham group on day 14 and day 28; and that of bronchoalveolar lavage fluid (BALF) was highest on day 7 than those on day 1 and day 14, while no significant difference existed between BLM group and sham group on day 28. Serum MMP-2 level did not change from day 1 to day 28, while the level of BALF MMP-2 began to increase after day 14, even on day 28. Lung tissue homogenate MMP-2 level began to increase early on day 3 and continued to day 28. CONCLUSION: The sources and effects of MMP-2 and MMP-9 differ in BLM-induced rat pulmonary fibrosis. PMID- 17121241 TI - [Virological response of chronic hepatitis C management with peg-interferon alpha 2b and ribavirin]. AB - OBJECTIVE: To evaluate the virological response of managing chronic hepatitis C (CHC) with peg-interferon alpha-2b (PEG-IFN alpha-2b) and ribavirin. METHODS: We retrospectively analyzed the virological response of 40 patients with different genotypes of hepatitis C virus (HCV) infection after anti-HCV management. Patients were given different dosages of PEG-IFN alpha-2b and ribavirin based on their weights. The duration of treatment was 48 weeks for patients infected by HCV genotype 1, and was 24 weeks for the others. HCV RNA was tested before treatment, 12 weeks post management, end of treatment, and 24 weeks after treatment stopped. RESULTS: Data from 40 patients were collected. Among them, 24 cases experienced HCV genotype 1 infection, and 16 cases were infected with other genotypes. Between these two groups, the early virological responses were 75.0% (18/24) and 87.5% (14/16), the end-of-treatment virological responses were 80.0% (16/20) and 85.7% (12/14), and the sustained virological responses were 56.2% (9/16) and 78.5% (11/14), respectively. CONCLUSION: Body weight-based customized PEG-IFN alpha-2b in combination with ribavirin can effectively treat patients with different genotypes of CHC. PMID- 17121242 TI - [Advances in primary thyroid lymphoma]. AB - Primary thyroid lymphomas (PTLs) are closely correlated with the autoimmune reaction of thyroid. However, the molecular mechanisms of PTLs are still unclear. It is really necessary to improve the ability to differentiate between benign and malignant PTLs along with the introduction of some new molecular biology methods. The diagnosis and prognosis of PTLs depend on their histological features, pathological classification, and clinical stages. Customized therapy of PTLs becomes possible with the further advances in lymphoma's pathological classification, clinical stages, and international prognosis index standard. PMID- 17121243 TI - [Clinical features and treatment options for aggressive angiomyxoma]. AB - OBJECTIVE: To discuss the clinical features and treatment options for aggressive angiomyxoma (AAM). METHODS: We retrospectively analyzed the clinical data of 4 patients with AAM treated at PUMC Hospital from January 1990 to December 2004. RESULTS: There were 1 man and 3 women with an average age of 34 years. The average age of the female patients were 27 years. Two patients (50%) had urinary or enteric compressive complaints, while another 2 patients had no clinical symptoms. Two patients underwent transvaginal surgeries, 1 patient underwent transabdomenal surgery, and 1 patient underwent subcurtaneouly local excision. Three patients (75%) experienced recurrences after operation and the median relapse time was 2.5 years. CONCLUSIONS: AAM usually occurs in female pelvic cavity and the soft tissues of female perineum. AAM is huge in capacity, and easy to infiltrate or recur locally. PMID- 17121244 TI - [Prevention and screening are always a priority and in the agenda in gynecology obstetrics...]. PMID- 17121245 TI - [3D and 4D imaging in fetal echocardiography]. AB - Cardiac defects form the most frequently encountered group of congenital malformations (0.8%). Despite the improving ultrasonographic technology and the teaching of the investigators, the prenatal recognition of cardiac defects stays low (around 30%). The application of 3D technology in their recognition may well lead to a substantial improvement of this percentage. This technique consists of a consecutive summation of multiple 2D images and permits the three dimensional reconstruction of the different structures and the definition of their spatial position. Its importance is localised in the direct identification of normal anatomy and the possibility to reevaluate the data in an off-line version and if necessary to ask for a specialised second opinion on the same data set, thus opening the door for telemedical approach. PMID- 17121246 TI - [Prevention of preterm delivery: new interventions]. AB - Prematurity is an important cause of perinatal mortality and morbidity. The incidence is not decreasing, despite the efforts of prevention. Few interventions were demonstrated effective to decrease the risk of preterm birth. We are currently conducting two randomized trials including women with preterm labor. The objective of one trial is to evaluate a psychotherapeutic intervention. The other trial is multicentric to evaluate vaginal progesterone. We hope that these two trials will provide us with valid evidence on the effectiveness of these interventions to decrease the risk of preterm delivery and of neonatal consequences. PMID- 17121247 TI - [Prophylactic HPV vaccine: "a giant leap for mankind and..." questions for our society!]. AB - Cervical cancer remains a major public health problem in the world. The strong link between human papillomavirus (HPV) and cervical cancer brings us the opportunity to develop a prevention against HPV infection through vaccination. HPV 16 and 18 induce about 70% of all cervical cancer, so prophylactic vaccine must be focus on these types. Trails established a great safety of theses vaccines and the induction of high titer of antibodies. They provide about 100% of protection against persistent HPV 16/18 infection and associated high-grade dysplasia. HPV vaccine must be proposed to young women "HPV-naive" before beginning of their sexual activity. But questions arise for our society: when to perform it, who would be on charge, inclusion of boys also, parental approval, adaptation of cervical screening and high cost. PMID- 17121248 TI - [Should we resect all premalignant breast lesions?]. AB - Premalignant breast lesions are defined by their characteristic histologic features. Based on these criteria, an increased risk is assigned of eventually progressing towards a malignant lesion, but the most majority of these lesions will never progress to cancer. Hence, the clinical management depends on correct sampling and thorough histologic examination. PMID- 17121249 TI - [Sacral staged reflexes to localize the pudendal compression: an anatomical validation of the concept]. AB - Pudendal neuropathy is common. The diagnosis is clinical and the confirmation is electrophysiological. Distal pudendal nerve latencies have been used but they are unspecific and do not allow to localize the site of compression. A preliminary electrophysiological study has suggested separate innervations of the anterior and the posterior anal sphincter quadrants, so diverging from what main anatomy textbooks teach. By detailed dissections of pudendal nerve region we can confirm a dichotomy in the innervation of the two quadrants. Therefore, it seems feasible, by using the differences of staged sacral reflexes, to better localize the compressive neuropathy, with a stimulation of the clitoris and by recording latencies of different muscles. PMID- 17121250 TI - [Laparoscopic treatment of genital prolapse]. AB - Progresses performed in laparoscopic surgery during the last ten years offer the possibility to do complex and difficult gynaecologic operations by laparoscopy, specially surgical treatment of genital prolapse, with interesting results. We report the main laparoscopic techniques to treat genital prolapse. It is clear that most of the laparoscopic procedures are the same than the classical operations performed by laparotomy. The main techniques that we report treat completely the pelvic organ prolapse (POP), without any vaginal scar, avoiding the risks of the dyspareunia in the sexually active patients. PMID- 17121251 TI - [Patient waiting time at the Medical and Surgical Emergency Center at the Hopital des Cadolles: a descriptive study]. AB - Analyzing how much time patient are waiting in the Medical and Surgical Emergency Center (Centre medico-chirurgical des urgences, CMCU) at the Hopital des Cadolles (Neuchatel, Switzerland) before discharge, over a 80 days period found that overall, patients spent on average 2 h 26 in the CMCU. There was a difference between hospitalized patients (3 h 16) and discharged patients (1 h 48). The time from first encounter with a health care provider until discharge or admission was 1 h 42 on average. Again, this time was longer for hospitalized (2 h 15 versus 1 h 17 for ambulatory patients). The majority of patients (76%) were assessed by a nurse or a physician within 5 minutes, while the other 24% of patients had to wait 15 minutes on average. Specifically, 26% of patients were seen by a physician within 5 minutes, while the other 74% had to wait 26 minutes on average. PMID- 17121252 TI - [Why not kill one's mother-in-law?]. PMID- 17121253 TI - [Iraq: when George W. Bush strikes at The Lancet]. PMID- 17121254 TI - [Pregnancy and nicotine substitution treatment: what to do?]. PMID- 17121255 TI - [The giddiness of infanticides]. PMID- 17121257 TI - [The jarring notes of the Swiss health system]. PMID- 17121256 TI - [Dogs recover their sight thanks to viral vectors that could be used in humans]. PMID- 17121258 TI - Relapse is different in recovery. PMID- 17121259 TI - No time to reinvent the wheel. PMID- 17121260 TI - Inspiring change leaders. PMID- 17121261 TI - Advice for new leaders. PMID- 17121262 TI - What's going on with meth? PMID- 17121263 TI - The potential role of tailored messaging. PMID- 17121264 TI - Common board problems--and solutions. PMID- 17121265 TI - Awake, hungry, and stressed. PMID- 17121266 TI - Increasing scrutiny of antipsychotics. PMID- 17121267 TI - When the computer offers advice. PMID- 17121268 TI - Improving our workforce. PMID- 17121269 TI - Structure and nomenclature of inositol phosphates, phosphoinositides, and glycosylphosphatidylinositols. PMID- 17121270 TI - Inositol and plant cell wall polysaccharide biogenesis. PMID- 17121271 TI - Functional genomics of inositol metabolism. PMID- 17121272 TI - Genetics of inositol polyphosphates. PMID- 17121273 TI - Inositol in bacteria and archaea. PMID- 17121274 TI - Regulation of 1D-myo-inositol-3-phosphate synthase in yeast. PMID- 17121275 TI - The structure and mechanism of myo-inositol-1-phosphate synthase. AB - The first and rate-limiting step in the biosynthesis of myo-inositol is the conversion of D-glucose 6-phosphate to 1L-myo-inositol 1-phosphate catalyzed by 1L-myo-inositol 1-phosphate synthase (MIP synthase). MIP synthase has been identified in a wide variety of organisms from bacteria to humans and is relatively well-conserved throughout evolution. It is probably homotetrameric in most if not all cases and always requires NAD+ as a cofactor, with NADH being reconverted to NAD+ in the catalytic cycle. This review focuses on the structure and mechanism of MIP synthase, with a particular emphasis on the mechanistic insights that have come from several recent structures of the enzyme. These include the structure of the enzyme from Saccharomyces cerevisiae, Archeoglobus fulgidus and Mycobacterium tuberculosis. PMID- 17121276 TI - Phosphoinositide metabolism: towards an understanding of subcellular signaling. PMID- 17121277 TI - Cracking the green paradigm: functional coding of phosphoinositide signals in plant stress responses. PMID- 17121278 TI - Inositols and their metabolites in abiotic and biotic stress responses. PMID- 17121279 TI - Inositol phosphates and phosphoinositides in health and disease. AB - In the past two decades, considerable progress has been made toward understanding inositol phosphates and PI metabolism. However, there is still much to learn. The present challenge is to understand how inositol phosphates and PIs are compartmentalized, identify new targets of inositol phosphates and PIs, and elucidate the mechanisms underlying spatial and temporal regulation of the enzymes that metabolize inositol phosphates and PIs. Answers to these questions will help clarify the mechanisms of the diseases associated with these molecules and identify new possibilities for drug design. PMID- 17121280 TI - Mammalian inositol 3-phosphate synthase: its role in the biosynthesis of brain inositol and its clinical use as a psychoactive agent. PMID- 17121281 TI - Evolutionary divergence of L-myo-inositol 1-phosphate synthase: significance of a "core catalytic structure". PMID- 17121282 TI - Plasmepsin II inhibitory activity of alkoxylated and hydroxylated chalcones. AB - Sixteen antimalarial alkoxylated and hydroxylated chalcones were tested for their ability to inhibit recombinant plasmepsin II in vitro. The best inhibitory compounds had either a chloro or dimethylamino group at the 4-position of phenyl ring A in the chalcone template. Combination of the chalcones with chloroquine showed additivity or mild antagonism of Plasmodium falciparum K1 growth in culture. PMID- 17121283 TI - Efficacy and safety of halofantrine in Pakistani children and adults with malaria caused by P. falciparum and P. vivax. AB - One hundred two patients aged 2-43 years diagnosed with acute malaria due to P. falciparum or P. vivax were treated with 3 doses of halofantrine (500 mg for > or = 18 year old patients and 8 mg/kg of patient body weight for 2-17 year olds), with each dose administered once in 6 hours and followed up for 28 days. Out of 102 patients 63 had P. falciparum, 36 had P. vivax and 3 had unidentified species. Following three dose therapy, 96.1% (98/102) of patients were cured, 0.98% (1/102) showed improvement from baseline, 1.96% (2/102) did not respond and were considered as treatment failures and one patient had indeterminate data. The lone patient, who relapsed after 120 hours post dose 1, was cured following re treatment on day 7. The median parasite clearance and fever clearance times, from the first dose, were 26 hours and 30 hours, respectively. Eleven point eight percent (12/102) of patients reported adverse events, of which abdominal pain, reported by one subject, was considered to be probably related to the drug and required corrective therapy. There were no serious adverse events or fatalities and none of the patients had a change in QTc interval greater than 10%. Thirteen point seven percent (14/102) of patients had abnormal clinical laboratory parameters that normalized later. PMID- 17121284 TI - Antimalarial activity of concanamycin A alone and in combination with pyronaridine. AB - Concanamycin A, a macrolide antibiotic inhibitor of vacuolar H+-ATPase derived from Streptomyces sp, inhibited Plasmodium falciparum K1 growth in culture with an IC500 value of 0.2 nM. It exhibited an additive effect when tested together with the antimalarial pyronaridine. PMID- 17121285 TI - Modern medicine and indigenous health beliefs: malaria control alongside "Sadsana phee" (animist belief system) in Lao PDR. AB - The major religion in Lao PDR is Buddhism, but many ethnic groups in rural Lao PDR hold an animist belief system called "Sadsana-pee". At the same time, the Bourapar District study site in Khammouane Province, Lao PDR is at high risk of malaria infection. Due to their belief in traditional ways of healing, the promotion of malaria prevention and treatment with modern medicine was not always welcomed by the villagers. Based on the results of questionnaire interviews with 240 heads of households from February to March of 2003, the effect of local beliefs on malaria control activities was discussed. Despite widely available western medicine and widely conducted health education, some people still believe that evil spirits cause malaria and rely on traditional medicine and/or religious ceremonies for treatment. Based on our findings, we recommend that future education and malaria control programs be revised and made sensitive to those people holding indigenous beliefs. PMID- 17121286 TI - Distribution of mast cells in relation to Schistosoma japonicum induced lesions in pigs. AB - The pathogenesis of schistosomiasis japonica has been extensively studied, however only little attention has been paid to the presence and localization of mast cells in relation to Schistosoma japonicum induced lesions. The aim of the present pilot study was to assess the parasitological and pathological responses in S. japonicum infected pigs with emphasis on the description of the distribution of mast cells in relation to lesions in the liver and cecum. Six pigs were exposed to 2,000 cercariae and examined 9 weeks post-infection. Three unexposed pigs of the same age served as helminth free controls. All infected pigs developed granulomatous hepatitis and typhlitis. In the liver, the degree of mast cell infiltration was higher in the infected pigs compared to the unexposed control group. This distinction could not be shown in the cecum. In both the liver and cecum, a mild to moderate number of mast cells were present within the granulomas. A significant relation was found between infection with S. japonicum and the mast cell infiltration in the liver. Due to their possible association with hepatic fibrosis, it seems as if they have some function in the fibrogenic process and thereby play a dual role in the pathogenesis of S. japonicum. In conclusion, the results show that mast cells are recruited to egg induced lesions in both the liver and the cecum. PMID- 17121287 TI - Strongyloidiasis with gastric mucosal invasion presenting with acute interstitial nephritis. AB - We report an atypical occurrence of invasive Strongyloides stercoralis infection of the stomach mucosa in an elderly female patient from Bangka Island, northwestern Indonesia. The patient presented with severe epigastric pain, edema of the legs, proteinuria and severe hypoalbuminemia. Gastric and duodenal biopsies found eggs, larval and adult forms present in the superficial mucosa with mild inflammation. The Harada-Mori filter paper culture technique revealed S. stercoralis filariform larvae and free-living adult worms, corroborating the diagnosis. The infection was associated with acute interstitial nephritis. The patient showed rapid and dramatic improvement after treatment with mebendazole. PMID- 17121288 TI - Acute cholecystitis caused by Clonorchis sinensis. AB - We are reporting a case of a 43-year-old Chinese male from Hong Kong, who came to see a doctor complaining of acute onset of severe upper abdominal pain. A diagnosis of acute cholecystitis was made and an emergency cholecystectomy was carried out. On opening the common bile duct, lancet-shaped worms were seen emerging from it. About 45 adult worms were collected and sent to the Department of Parasitology University of Malaya for identification. The worms were identified as Clonorchis sinensis. After the operation the patient was treated with praziquantel and he had an uneventful recovery. PMID- 17121289 TI - Eye lesion caused by adult Brugia malayi: a first case reported in a child from Malaysia. AB - We are reporting a case of an eye lesion caused by an adult Brugia malayi. The patient was a 3-year-old Chinese boy from Kemaman District, Terengganu, Peninsular Malaysia. He presented with a one week history of redness and palpebral swelling of his right eye. He claimed that he could see a worm in his right eye beneath the conjunctiva. He had no history of traveling overseas and the family kept dogs at home. He was referred from Kemaman Hospital to the eye clinic of Hospital Tengku Ampuan Afzan, Kuantan, Pahang, Malaysia. On examination by the ophthalmologist, he was found to have a subconjunctival worm in his right eye. Full blood count revealed eosinophilia (10%). Four worm fragments, each about 1 cm long were removed from his right eye under general anesthesia. A thick blood smear stained with Giemsa was positive for microfilariae of Brugia malayi. A Brugia Rapid test done was positive. He was treated with diethylcarbamazine. PMID- 17121290 TI - Laboratory colonization of Mansonia mosquitoes with an emphasis on Ma. annulata and Ma. bonneae. AB - The present study records the first successful colonization of Mansonia annulata and describes colony maintenance with modification of rearing medium and host plants. Three species of Mansonia mosquitoes (Ma. uniformis, Ma. indiana and Ma annulifera) were successfully reared in ambient environments with adult emergence rates > 50%, while Ma. bonneae and Ma. dives yielded emergence rates > 30%. Colonization of Ma. annulata was modified and improved so that they were successfully raised to adult with emergence rates of 23%. Tube sedge, Lepironia articulata, was utilized as a host plant and peat swamp water was used as a rearing medium. Yeast and small lizard droppings were added daily to the larval medium to maintain microorganisms and pH in the infusion. However, identifying suitable culture medium remains an obstacle to establishing colonies of Ma. annulata, as the culture medium is difficult to mimic in the laboratory. Further study, focusing particularly on larval attachment substrates and rearing medium, is needed to develop a standardized and practical rearing technique for Mansonia mosquitoes. PMID- 17121291 TI - Field investigation of Bithynia funiculata, intermediate host of Opisthorchis viverrini in northern Thailand. AB - A survey of freshwater snail, Bithynia funiculata, was conducted in four locations, Doi Saket, Mueang, Saraphi and Mae Rim Districts of Chiang Mai Province, northern Thailand, between June and October, during the rainy season of 2004. A total of 2,240 snails was collected and classified into 7 families and 15 genera; of which 352 B. funiculata were obtained. B. funiculata was found most abundant in July and September. The infection rate of trematode cercariae in B. funiculata was 9.6% (19/352), while that of pleurolophocercous was 0.3% (1/352). Virgulate cercaria was the most common type, followed by lophocercous, monostome and pleurolophocercous. B. funiculata prefers habitats with clear water, which was less than 30 cm depth, temperatures between 24.48 and 31.78 degrees C, dissolved oxygen 2.03-7.66 mg/l, saturated dissolved oxygen 26.70-95.00%, conductivity 0.000-0.2642 mS/cm, turbidity 16.00-288.00 NTU and pH 6.58-7.56. Geographic Information System (GIS) analysis using loose soil mixed with clay revealed that the breeding grounds of this snail species was in the paddy fields and village environs of the Ping, Kuang and Fang river basins. PMID- 17121292 TI - Analysis of a previous smallpox vaccination study: estimation of the time period required to acquire vaccine-induced immunity as assessed by revaccination. AB - The time interval required to develop immunity after vaccination, in the event of a bioterrorist attack using variola virus, is yet to be clarified. In this article, a historical vaccination study conducted in Japan in 1929 was re examined. Forty-four previously vaccinated and 44 unvaccinated children were involved. After successful first round primary (or re-) vaccination, all children underwent revaccination at variable intervals. Absence of a major reaction (vaccine 'take') after revaccination was taken as a sign of immunity conferred by first round primary (or re-) vaccination. Univariate analysis was employed to examine the relationship between vaccine 'take' and the exposure variables. Maximum likelihood estimates of the time period required to develop immunity were obtained using a simple logit model. The interval between vaccinations was significantly associated with vaccine 'take' in both the previously unvaccinated (p < 0.01) and vaccinated (p < 0.01) groups, and the median interval required for immunity after vaccination was estimated to be 6.4 [95% Confidence Interval (CI): 5.8, 7.1] and 4.3 days (95 % CI: 4.1, 4.7), respectively. Obtained estimates were consistent with previous observations, and the logistic fits reasonably explained the discrepancy among previous suggestions. The findings suggest that it is necessary to vaccinate exposed susceptible individuals within 3 days after exposure to ensure disease protection, and within at least 5 days (for those previously unvaccinated) to provide a certain level of protection; the probability shows a dramatic decline hereafter. PMID- 17121293 TI - Dengue in pregnancy. AB - This was a retrospective study of patients with dengue infection in pregnancy from year 2000 till 2004. Data were analyzed by looking at the presentation, complications of patient and fetus, and pregnancy outcomes. There was a total of 16 cases with an increasing trend (0.12% in 2003 vs 0.25% in 2004). The mean age of patients was 30.19+/-6.85 years. Fifty percent of patients were multiparae and in their third trimester. The average gestation was 24.44 weeks with 7.5 days being the average duration of ward admission. Tourniquet test was positive in 62.5% of patients. Dengue serology IgM was positive in 50% whereas dengue serology IgG were positive in 68.8% of patients. There were three cases of maternal death. One patient presented as missed abortion. Preterm deliveries happened in 50.0% of the women. There were 4 premature babies, one in-utero fetal death, and two fetuses which suffered acute fetal distress. Three babies required intensive care. One unrelated fetal anomaly resulted in early neonatal death. PMID- 17121294 TI - Avian influenza A (H5N1) infection in a child. AB - A previously healthy, 9-year-old girl was admitted to the hospital with respiratory insufficiency. She had mild and severe respiratory symptoms for 3 weeks and 4 days before admission, respectively. She had a history of close contact with her domestic poultry, which was infected with avian influenza A (H5N1). She was isolated with the air-borne transmission prevention mode of treatment. Acute respiratory distress syndrome (ARDS) was documented from the time of admission and mechanical ventilation was introduced without improvement. She had multiple episodes of diarrhea for 2 days. Her condition deteriorated and she expired in 4 days. Throat swab RT-PCR and viral culture for avian influenza A (H5N1) were positive. PMID- 17121295 TI - Factors associated with non-disclosure of HIV infection status of new mothers in Bangkok. AB - The objective of this study was to estimate HIV disclosure rates and identify factors that predict non-disclosure in Thai women who tested HIV positive during pregnancy or at delivery. This was a cohort study evaluating the implementation of prevention of mother-to-child HIV transmission programs at two Bangkok hospitals in 1999-2003. All HIV-infected women who delivered during the study period were enrollment eligible. Thai-language questionnaires were used to collect baseline data before discharge from the hospital. At the 1 and 4 month follow-up visits, women were asked if they had disclosed their HIV status. Of the 799 women who enrolled, 647 (81.0%) completed follow-up at 1 and 4 months. Four hundred fifty-three (70.0%) women disclosed their status by 1 month. Of the 194 women who had not disclosed by 1 month, 48 (24.7%) had disclosed their status by 4 months. An independent increased odds of non-disclosure by 1 month was associated with not having a partner tested for HIV (OR=5.83, 95% CI=3.19-9.08) or not knowing if the partner was ever tested for HIV (OR=1 3.02, 95% Cl=5.26 32.28), first learning of HIV positive status during delivery (OR=6.84, 95% CI=2.36-19.81) or after delivery (OR=3.14, 95% CI=1.57-6.26) and having >2 lifetime sexual partners (OR=1.71, 95% CI=1.04-2.82). Not living with a partner every day was associated with non-disclosure by 4 months in those women who had not disclosed by 1 month (OR=2.28, 95% CI=1.43-3.64). Despite high rates of disclosure by 1 month, 22.6% of women still had not disclosed their HIV status to their partners by 4 months. The benefits of disclosure warrant effective interventions targeted at women at risk for non-disclosure. PMID- 17121296 TI - The National Access to Antiretroviral Program for PHA (NAPHA) in Thailand. AB - To describe the development, components, initial results and lessons learned from Thailand's National Access to Antiretroviral Program for People living with HIV/AIDS (NAPHA), a historical review was conducted and program monitoring was analyzed. The national antiretroviral therapy program at different levels of the public health system was implemented with all major program components; ARV protocol development, health care professional training, drug supply chain management, laboratory network formation, monitoring and evaluation, and multi sector and PHA involvement since 2001, which was based on elements of research, pilot projects, training, national guideline development, experiences and policy making. A national monitoring system was developed to monitor the progress of the program. From February 2001 to December 2004, the monitoring reports received from implementing hospitals showed that 58,133 cases had received antiretroviral therapy (ART), and 85% (49,477) of them were continuing to take ARV drugs. In conclusion, the NAPHA was implemented nationwide with comprehensive systems. The reports indicate achievement of expansion of the ART program. Lessons learned from the program initiation and scaling up show local leadership, comprehensive training, adherence, and coordination are essential to program effectiveness and sustainability. PMID- 17121297 TI - A serological investigation of leptospirosis in sows from central Thailand. AB - Leptospirosis is a major pig-associated zoonosis worldwide. There are also human occupational disease exposure risks. For better public health and sanitation, the prevalence of this disease was investigated. Four hundred sera were obtained from sows in central Thailand during August 2004 to January 2005 and examined with 20 reference leptospira serovars by a microscopic agglutination test (MAT). The results showed that 10% of samples gave a positive reaction. The most prevalent serovar was grippotyphosa which accounted for 55% of all positive cases. Other minor serovars were canicola and a non-pathogenic strain (patoc). It follows that, the sow may be one of the disease transmitting sources for humans in this area. The disease correlation between pigs, humans and the environments should be evaluated. PMID- 17121298 TI - A histopathological study of hearts and spleens of hamsters (Mesocricetus auratus) infected with Leptospira interrogans, serovar pyrogenes. AB - The effects of Leptospira interrogans on the heart and spleen of hamsters were studied histopathologically. Infected hamsters were sacrificed at 1 hour, 6 hours and on days 1, 2, 3, 4, 5 and 6 after inoculation with Leptospira interrogans serovar pyrogenes. The heart and spleen of each of the sacrificed animals were removed and processed for routine conventional light microscopy. Infected hearts showed degenerative change of the cardiac muscle cells composed of cellular swelling, condensation of chromatin granules, pyknotic nuclei and acidophilic cytoplasm. Congestion of the cardiac blood vessels and hemorrhagic areas were found. Necrosis of the cardiac muscle cells was surrounded by numerous inflammatory cells. In the spleen, cellular necrosis was found scattered throughout the splenic cord. The splenic sinusoids were dilated and congested with many hemorrhagic areas. Inflammatory cell infiltration was also noted in the splenic parenchyma and the splenic sinusoids. PMID- 17121299 TI - Duplex PCR-hybridization based detection of pathogenic Leptospira in environmental water samples obtained from endemic areas in northeast region of Thailand. AB - Leptospirosis, a major health problem worldwide, is known to be endemic in the northeastern part of Thailand with the risk of infection by exposure to pathogenic Leptospira in contaminated aquatic environment. A method based on PCR hybridization detection of pathogenic Leptospira in water was established. The method included filtration of water sample through membrane filters of two pore sizes, DNA extraction from filters using a guanidine thiocyanate extraction method, a duplex-PCR assay with two primer pairs, and hybridization with a synthetic LipL32 DNA probe. The duplex-PCR allowed detection of two products of 279 bp for LipL32 gene and 430 bp for 16S rRNA gene. In water samples artificially seeded with serovar bratislava, at least 10(3) cells could be analyzed by PCR-agarose gel electrophoresis and 1-10 cells by PCR-Southern blot hybridization. The protocol was applied to the detection of pathogenic Leptospira in environmental waters collected from endemic areas in the northeast region of Thailand. Of 100 water samples analyzed, 23 samples were positive for pathogenic Leptospira with PCR performed with Southern blot hybridization only, but none was detected by PCR-agarose gel-electrophoresis. However, PCR performed with the chemiluminescent LipL32 probe using the Fluorescein ULS labeling facilitated the detection of low numbers of pathogenic Leptospira in water. This method should prove useful for monitoring of pathogenic Leptospira pollution in environmental waters, and has the potential to become a valuable tool to the surveillance of leptospirosis in endemic areas, thus leading to enhanced public health protection. PMID- 17121300 TI - Recovery of Salmonella using a combination of selective enrichment media and antimicrobial resistance of isolates in meat in Thailand. AB - From November 2004 to March 2005, 50 samples (chicken, pork and beef) of registered meat and non-registered meat were purchased from supermarkets and retail markets located in Bangkok, Thailand. Each sample was evaluated for Salmonella spp by a conventional method using combination of selective enrichment media (RV+MSRV) and compared with selective enrichment medium alone (DIASALM). Our study revealed the performance of RV+MSRV for the detection of Salmonella spp was significantly better than those of DIASALM alone since the recovery of Salmonella spp in both groups of meat was high using RV+MSRV, particularly in the registered meat. In addition, the recovery of serovars in registered meat was significantly higher than those in non-registered meat. Antimicrobial resistance of 62 Salmonella isolates in both groups of meat was determined for 10 antimicrobial drugs using the disk diffusion method. The results show that 100% of isolates from both groups were susceptible to amoxicillin/clavulanic acid, ciprofloxacin, cefotaxime and norfloxacin and 50-60% of isolates from both groups were resistant to tetracycline, streptomycin and ampicillin. Sixty percent of Salmonella isolates from meat showed multiresistance antimicrobial patterns. PMID- 17121301 TI - Analysis of fecal leukocytes and erythrocytes in Shigella infections in urban Bangladesh. AB - We evaluated the usefulness of enumeration of fecal leukocytes and erythrocytes in making an early diagnosis of Shigella infection, where Shigella is a leading cause of invasive diarrhea. Stool specimens from 561 invasive diarrhea patients were submitted for microscopic examination. A presumptive diagnosis of shigellosis based on microscopic examination was made in 389 of them; 227 had stool cultures positive for Shigella spp (Shigella patients). One hundred sixty two patients with no detectable Shigella infection (non-Shigella invasive diarrhea cases) served as a comparison group. Two hundred twenty-seven randomly selected Shigella patients and 227 non-Shigella infectious diarrhea cases from the surveillance system database of the hospital constituted another group for comparative evaluation. The stool specimens of the patients were examined under the microscope, and isolation, biochemical characterization and serotyping of Shigella were performed. In comparison with non-Shigella invasive diarrhea cases, the presence of >50 WBC/hpf in association with any number of RBC in the fecal sample had a modest sensitivity of 67%, specificity of 59%, positive predictive value of 70%, negative predictive value of 56%, accuracy of 64%, and positive likelihood ratio of 1.6 in predicting shigellosis. Comparison between Shigella and non-Shigella infectious diarrhea patients revealed the presence of >20 WBC/hpf was a less accurate predictor of shigellosis (sensitivity 51%, specificity 88%, positive predictive value 81%, negative predictive value 64%, accuracy 69%, and positive likelihood ratio 4.1). Direct microscopical examination of stool specimens for the presence of WBC and RBC may facilitate the early diagnosis of shigellosis, and may be a cheap alternative to stool culture in this setting. PMID- 17121302 TI - Surat plague of 1994 re-examined. AB - A plague episode in Surat in 1994, and its spread to other cities in India, lasted only a little over 2 weeks, but it created an unprecedented panic that had global repercussions. At first, the Surat hospital doctors could not diagnose the disease, but when they did, immediate intervention, in the form of preventation and treatment (administration of antibodies) prevented the disease from spreading beyond Surat, Delhi, Calcutta, Bombay and their vicinities. Fewer than 1,200 people were diagnosed with plague. A DNA-based study in 2000 decisively concluded that the Surat episode was a plague, but the Indian isolates were genetically more heterogeneous compared to others in the world. PMID- 17121303 TI - Detection of heterogeneous, intermediate-vancomycin-resistant Staphylococcus aureus (hVISA) using low-concentration vancomycin disks. AB - Heterogeneous, intermediate-vancomycin-resistant Staphylococcus aureus (hVISA) represents a threat of an incurable infection since the first report in 1997. The method used to detect hVISA isolates is a population analysis profile (PAP); however, it is impractical for routine laboratory analysis. We therefore tested a simple, reliable and inexpensive method for the detection of hVISA. Eighteen isolates of hVISA and 22 of vancomycin-sensitive S. aureus (VSSA) were included. The organisms were tested by the disk diffusion method, using 15-microg vancomycin disks on four different media: Mueller-Hinton agar (MHA), MHA plus 2% NaCI (MHAS), Brain Heart Infusion agar (BHA), and BHA plus 2% NaCl (BHAS). In addition, two different inoculum sizes, bacterial suspensions adjusted to 0.5 and 2.0 McFarland, were tested. The inhibition zone was read independently by three medical technologists after incubation at 37 degrees C for 24 and 48 hours. The use of MHAS with an inoculum size of 2.0 McFarland and 48-hour incubation period yielded the highest sensitivity (94.4%), specificity (81.8%), positive predictive value (80.9%), and negative predictive value (94.7%). The disk diffusion test with 15-microg vancomycin disk is simple and may be used as a screening method for the detection of hVISA. PMID- 17121304 TI - Detection of Cryptococcus neoformans in bird excreta. AB - We evaluated 14 samples of bird excreta from pigeons, parrots, open billed storks and crows obtained from thirteen places in Bangkok and nearby areas between April and July 2004. These bird excreta were examined for Cryptococcus neoformans by direct plating method to inspect their ability to grow at 37 degrees C. Capsule production was examined by Indian ink preparation. They were also tested for urease and phenoloxidase enzymes. Cryptococcus neoformans var neoformans was recovered from pigeon excreta in 9.09%. This implies those having impaired immunity may get this fungus from the environment. PMID- 17121305 TI - Childhood anemia and vitamin a deficiency in rural Bangladesh. AB - The study aimed to assess the prevalence of anemia and vitamin A deficiency in preschool children in rural Bangladesh. A cross-sectional study was done on eight randomly-selected sub-districts of rural Bangladesh. Children (n=1,302) aged 2-6 years were studied. Families of 43% of the study participants had a monthly household expenditure of US$ 60 or less. Fifty-six percent of the children were underweight, and 17% were severely underweight; 18% were wasted, and 1% were severely wasted; and 45% were stunted while 20% were severely stunted. The mean+/ SD serum retinol of the children was1.0+/-0.4 micromol/l, and 3% of them had serum retinol levels of <0.35 micromol/l, about one-fifth (20%) had a serum retinol level of <0.70 micromol/l and 55% had serum retinol levels of <1.05 pmol/l. The mean hemoglobin concentration of the children was 110+/-11 g/l, and 48% had a Hb of <11 g/l signifying anemia in this age group. Thirty-one percent (31 %) of children had low serum ferritin (<12 microg/l), and 14% had elevated CRP (> or = 15 mg/l) indicating the presence of a sub-clinical infection. Male and female children had similar nutritional status and biochemical profiles although boys tended to be heavier than girls (p=0.013). The proportion of children with anemia and iron deficiency anemia (IDA) declined significantly (p<0.001) with advancing age. Five percent of the study children had IDA and concomitant low serum retinol. The proportion of children with IDA and serum retinol also declined significantly with increasing age from 8% in children aged 35 months or younger, to 3% in children aged 60 months and more (p=0.025). Results of our study clearly demonstrated the public health importance of anemia and vitamin A deficiency among children of rural Bangladesh. PMID- 17121306 TI - Postprandial leptin response to Thai meals with different macronutrient mixtures. AB - The objective of this study was to investigate the postprandial response of leptin, an appetite-regulating hormone, to different macronutrient mixtures in Thai meals. A within-subject repeat measurement was performed. Two groups of healthy Thais (10 men and 10 women in each group) received a single meal of equal calories composed either a high carbohydrate, low fat, low protein diet (HC-LFLP, carbohydrate:fat:protein = 70%:15%:15%) or a low carbohydrate, high fat, high protein diet (LC-HFHP, carbohydrate:fat:protein = 20%:50%:30%). Fasting and 30 minute interval postprandial blood levels of leptin, insulin and glucose were measured for a 2-hour period. In comparison to the LC-HFHP meal, the HC-LFLP meal produced a greater increase in glucose and insulin levels, but halted leptin from decreasing. Postprandial leptin levels were suppressed by a LC-HFHP meal but not by a HC-LFLP meal. The reduced leptin in conjunction with lower glucose and insulin levels may encourage overeating in habitual LC-HFHP diet consumers. PMID- 17121307 TI - Anthropometric characteristics of elderly people: observations at a large diarrheal hospital in Dhaka, Bangladesh. AB - There is a lack of evidence-based information to assist health policy makers in preparing for appropriate health, nutrition, and social-support guidelines for the elderly in Bangladesh. We examined selected indicators of the nutritional status of elderly people attending the Dhaka Hospital of ICDDR,B, Dhaka, Bangladesh. The population constituted of 1,196 individuals (718 men and 478 women), aged 60 to 106 years, who attended the hospital between 1 January 1993 and 31 December 2003. Patients were recruited from a hospital-based systematic sampling, regardless of age and gender, that presented to the facility. Men were heavier, and taller than women were (p < 0.001 for both comparisons). Using MUAC cut-off of < 22 cm for females and < 23 cm for males, at least 50% of the elderly were peripherally wasted (malnourished). Among all the study population, 40% had a BMI within the optimal range (18.5-24.9 kg/m(2)). Using the chronic energy deficiency (CED) classification, at least half of elderly (> or= 60 year) women were chronic energy deficient (BMI < 18.5). A significantly higher proportion of elderly women (7%) compared to men (2%) were overweight (BMI > or = 25, p < 0.001). Among the elderly ( > or = 60 year), males and females from a higher socioeconomic status (SES) had significantly higher BMI (p < 0.001, p = 0.001, respectively) and MUAC values (p < 0.001, p < 0.001, respectively) than their less well-off SES counterparts. We consider that, although our data were not valid for assessing the country situation, they are still useful as baseline information for longitudinal studies and for highlighting the need for studies in other geographical locations and in other population groups. PMID- 17121308 TI - Rapid detection of apolipoprotein E genotypes in Alzheimer's disease using polymerase chain reaction-single strand conformation polymorphism. AB - Apolipoprotein E (APOE) gene on chromosome 19q13.2 is encoded by three common alleles designated as epsilon2, epsilon3 and epsilon4. In Alzheimer's disease (AD) the epsilon4 allele is over-represented and is considered to be a major genetic risk factor. Several methods have been developed to determine APOE genotypes. Among them, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) appears to be highly reliable. In this study, we improved the nonisotopic PCR-SSCP method for determining APOE genotypes in 42 cases of AD patients, 40 cases of non-AD dementia patients, and 49 cases of age-matched controls. DNA from the target sequence on APOE was amplified by PCR from peripheral blood genomic DNA. PCR products were electrophoresed in a non denaturing polyacrylamide gel and visualized by silver staining. We found that the epsilon4 allele had a significantly high frequency of occurrence in AD patients (33.3%) compared with age-matched controls (13.3%) (chi(2) = 10.43, p = 0.001) and non-AD dementia (10%) (chi(2) = 13.02, p<0.001) whereas the epsilon3 allele was of high frequency in non-AD dementia (90%) compared with age-matched controls (85.7%) and AD patients (66.7%). APOE epsilon4 homozygotes were found only in AD groups. On the other hand, the epsilon2 allele was found only in an age-matched control. This study confirmed that the APOE psilon4 allele is a risk factor in Thai AD subjects and that the PCR-SSCP method is a rapid and useful means of detecting the APOE genotype in AD. PMID- 17121309 TI - National response to neurological diseases in Malaysia: planning for the future. AB - The number of cases of neurological disease is expected to rise in the next 10 years, making this the second leading cause of morbidity and mortality after heart disease in Malaysia. The lack of human resources in the neurological field currently serving the Malaysian population may cause a deficiency in specialized care, especially in rural areas where neurological and neurosurgical care may be lacking. Thus, a resolve was made to increase the numbers of specialists by the Universiti Sains Malaysia (USM) with the help of the Ministry of Health of Malaysia. A study was made to evaluate the number of referral centers needed in strategic parts of Malaysia. Our calculation was based on service demands and operative procedures following the guidelines of the Association of British Neurologists (ABN) where 15 minutes of service time was equivalent to 1 unit. Based on 2 million population covered in the state of Kelantan by this University Hospital, 4.27 neurologists are needed to meet service demands with a consultant to population ratio (CPR) of 1:468,384, compared to 7.46 neurosurgeons, with a CPR of 1:268,097. According to the current service demands, one neurologist has to work more than 407 hours per year and one neurosurgeon 1,219 hours per year in our hospital. Hospitals with a larger catchment area would need to have more neurologists and neurosurgeons for optimal care in their area. Thus, more neurologists and neurosurgeons are needed to be produced, since the existing numbers are too small for quality care in Malaysia. PMID- 17121310 TI - A study of the factors related to intima-media thickness of the common carotid artery amongst rural middle age individuals in Hospital Universiti Sains Malaysia. AB - There is very little data regarding the factors related to intima-media thickness (IMT) of the common carotid artery in normal individuals in those with non insulin diabetes mellitus and perimenopausal women in Southeast Asian countries. Ultrasound imaging evaluating the carotid artery IMT in those with diabetes and those on hormone replacement therapy (HRT) was performed beginning in August 2000 for a period of nearly two years at the Department of Radiology, Hospital Universiti Sains Malaysia, Kubang Kerian, Kelantan, Malaysia. A total of 153 participants were included. Significant differences between the women on HRT and not on HRT were IMT and systolic blood pressure. When comparing those with non insulin dependent diabetes mellitus (NIDDM) and normal individuals, the significant differences were IMT, total cholesterol level, systolic blood pressure and diastolic blood pressure. IMT was high in those with NIDDM but not in those on HRT. Both those with NIDDM and those on HRT had associated dyslipidemia and systolic hypertension. PMID- 17121311 TI - Cost-benefit analysis of an intensive adverse product reactions monitoring program of inpatients in Thailand. AB - The objective of this study was to analyze the costs and benefits of the intensive adverse products reactions (APRs) monitoring program of inpatients in medical wards at Nakhon Ping Hospital, Chiang Mai, Thailand. The data were retrospectively collected from inpatients who had APRs during admission period from November 16, 2004 to March 31, 2005. Products included were drugs, electrolyte solutions, bloods and blood derivatives. Only direct medical costs were considered using provider's perspective. The results showed that there were 1,407 admitted patients during the study period. Adverse products reactions were found in 31 patients. Of those, three patients had two APRs. Therefore, a total of 34 APRs were found yielding the APRs incidence rate of 24 per 1,000 inpatients. Of the APRs found, 20 were reported after the symptoms had begun, but the remaining APRs were preventable. Drugs were the main causes for APRs (94.12%). Cost of intensive APRs monitoring was 1,426.37 US dollars and included 939.44 US dollars for labor costs, 12.5 US dollars for material costs, 29.07 US dollars for capital cost and 445.36 US dollars for APRs treatment and investigation. The program cost saving excluding the cost of hemodialysis was 3,090.85 US dollars. Net benefit was 1,664.48 US dollars and benefit to cost (B/ C) ratio was 2.17. When one time hemodialysis cost was included, cost saving increased to 4,040.85 US dollars. Net benefit and B/C ratio were 2,614.48 US dollars and 2.84 respectively. The results of sensitivity analysis represented that net benefit and B/C ratio were increased when duration of digoxin intoxication was prolonged. However, net benefit and B/C ratio were decreased when pharmacist labor cost was increased. Results indicate that intensive APR monitoring is a cost beneficial program and should be continuously implemented. PMID- 17121312 TI - Clinical effect of a triclosan containing dentifrice on gingivitis during pregnancy and post-partum. AB - One hundred forty pregnant women 3 months gestation were enrolled in a 9 month, double-blind clinical study to evaluate the effect of 0.3% triclosan/copolymer dentifrice on gingivitis, compared to a placebo dentifrice. Women were stratified into two balanced groups according to their baseline gingivitis scores. They then received oral prophylaxis and were assigned to use either a placebo or the triclosan dentifrice for the next 9 months. The 3, 5 and 9 month results of this study showed that the triclosan dentifrice provided statistically significant reductions in gingivitis of 19.73, 27.91and 38.45%, respectively, compared to the placebo dentifrice. PMID- 17121313 TI - Quantification of regional left ventricular systolic dysfunction in patients with coronary artery disease by pulsed Doppler tissue imaging. AB - OBJECTIVES: To analyze and compare quantitatively regional myocardial functional abnormalities of the left ventricle by pulsed Doppler tissue imaging (DTI) in patients with unstable angina (UA) and prior myocardial infarction (MI), and to explore the value of systolic velocity and time intervals in evaluating regional left ventricular systolic dysfunction. METHODS: Patients with coronary artery disease (CAD) were divided into UA (16 cases) and anterior wall MI (ant-MI, 21 cases) groups. Sixteen age-matched normal subjects served as the control group. The septal and lateral, anterior and inferior walls of the left ventricle were displayed, and basal and middle segments of each wall were selected for myocardial motion spectrum sampling. DTI parameters were: peak systolic myocardial velocity (s), regional pre-ejection period (PEP), time to peak of the systolic wave (Ts), regional ejection time (ET) and PEP/ET ratio. RESULTS: Compared with the control group, s was significantly lower in all segments in the ant-MI group, and in lateral and anterior segments in the UA group. It was even lower in the ant-MI than in the UA group, and in infarct compared with corresponding non-infarct segments in the ant-MI group. PEP and Ts were significantly longer in both the UA and the ant-MI groups. CONCLUSION: Not only s, but also PEP and Ts as measured by pulsed DTI are sensitive markers of regional left ventricular systolic dysfunction in patients with CAD; s and PEP may even indicate the severity of myocardial ischemia and aid in estimating the site of MI. PMID- 17121314 TI - Systemic lupus erythematosus and renal involvement: which role of citokines expression? AB - The role of cytokines in systemic lupus erythematosus (SLE) glomerulonephritis is extremely complex. Proinflammatory molecules, such as TNF, IL-6, IL-1 and IL-18 are upregulated, as are both Thl and Th2 cytokines, with different implications: the local effects may be different from the systemic immunoregulatory ones. Excessive T helper cell function is a hallmark of SLE and abnormalities of Th citokine profiles have been implicated in loss of immune tolerance, increased antogenic load, defective B cell suppression and a variety of clinical manifestations. For some cytokines, TNF and IL-18 in particular, the local proinflammatory ones may be more relevant to the disease. PMID- 17121315 TI - Focus on juvenile idiopathic arthritis according to the 2001 Edmonton revised classification from the International League of Associations for Rheumatology: an Italian experience. AB - OBJECTIVE: To classify a cohort of Italian patients categorized as affected with juvenile idiopathic arthritis (JIA) according to the revised 2001 Edmonton International League of Associations for Rheumatology (ILAR) criteria. METHODS: Eighty-five patients with JIA firstly framed depending on traditional criteria during the last ten years were reallocated according to the JIA revised criteria proposed in 2001 by ILAR in Edmonton. RESULTS: The revision consented to define the following distribution of patients: 28.2% systemic, 55.3% oligoarticular and 11.8% polyarticular forms; only one child was defined as having psoriatic arthritis, one child with enthesitis-associated arthritis and two with the undifferentiated form of JIA. DISCUSSION: The 97.6% of the recruited patients were strictly classified according to the Edmonton ILAR criteria, demonstrating a very low number of patients whose arthritis could not be assigned to any JIA category due to unfulfillment of the required criteria. PMID- 17121316 TI - Normal Weight Obese syndrome: role of single nucleotide polymorphism of IL-1 5Ralpha and MTHFR 677C-->T genes in the relationship between body composition and resting metabolic rate. AB - We have identified a subset of metabolically obese, but normal weight individuals, with potentially increased risks of developing the metabolic syndrome, despite their normal body mass index. We determined the relationship among body fat distribution, resting metabolic rate (RMR), total body water amount (%TBW), selected gene polymorphism on interleukin-15 receptor-alpha (IL 15Ralpha) and methylenetetrahydrofolate reductase 677C-->T (MTHFR 677C-->T), to distinguish normal weight obese (NWO) from nonobese with a normal metabolic profile and obese individuals. We analysed anthropometric variables, body composition by Dual energy X-ray Absorptiometry (DXA), RMR by indirect calorimetry, %TBW by bioimpedence analysis (BIA), MTHFR 677C-->T and IL-15Ralpha genotypes of 128 clinically healthy Caucasian individuals. We compared a group of female, defined as NWO and characterised by a BMI < or = 25 kg/m(2) and FM > or = 30% with groups of others female, and males, represented by nonobese with a BMI < or = 25 kg/m(2) and FM < or = 30%, and preobese-obese individuals with BMI > or = 25 kg/m(2) and %FM > or = 30%; none of the males was classified as NWO. Significant correlations were found among body fat mass distribution, metabolic variables, percentage of total body water distribution and selected genetic variations. The variables that contributed significantly to the separation of classes were body tissue (Tissue), %TBW, RMR, the volumes of both oxygen (VO2) and carbon dioxide (VCO2). The distribution of MTHFR 677C-->T and IL-15 genotypes was significantly different between classes. Our data highlight that NWO individuals showed a significant relationship between the decrease in the basal metabolism (RMR), body fat mass increasing and total water amount. Possession of wild type homozygotes genotypes regarding IL-15Ralpha cytokine and 677C-->T MTHFR enzyme characterised NWO individuals. PMID- 17121317 TI - Myoinositol/folic acid combination for the treatment of erectile dysfunction in type 2 diabetes men: a double-blind, randomized, placebo-controlled study. AB - Erectile dysfunction is a common complication of diabetes. Diabetes can cause neuropathy or damage to nerves throughout your body, including the penis. Damaged nerves can't communicate properly. So even though you might be emotionally stimulated to have intercourse, nerve damage means that information isn't relayed to the penis, and it doesn't respond. In addition, poor blood sugar control can inhibit nitric oxide production. Lack of nitric oxide can prevent the pressure of blood in the corpora cavernosa from rising enough to close off penile veins, allowing blood to flow out of the penis instead of remaining trapped for an erection. This prospective, randomized, double-blind, placebo-controlled study included 176 patients with type 2 diabetes. The daily 4 g dose of inositol plus 400 microg of folic acid or placebo was divided and given in three doses. The present study demonstrates that Myoinositol/folic acid combination, deserves consideration as therapeutic agent for preventing and treating erectile dysfunction in diabetic men, probably by virtue of both their chronic metabolic, acute ROS scavenging, and NO protective beneficial effects. PMID- 17121318 TI - Solitary plasmacytoma of the spine: a 22 years follow-up case report. AB - BACKGROUND AND OBJECTIVES: Plasmacytoma is a systemic malignancy and it is the most frequent primary tumor affecting the skeleton. Progression from solitary plasmacytoma to systemic diffusion is reported to be 65-100% in 15 years. A case report of a rare 22-year follow-up of a thoracic solitary plasmacytoma is here presented. MATERIAL AND METHODS: Clinical case analysis, radiographs, magnetic resonance images and histological sections of the lesion are discussed. Repeated surgical interventions were required due to progression and local recurrence of disease. RESULTS: Although the inadequate surgical treatment, low response to chemotherapy and radiation therapy, and many local recurrences, no systemic diffusion of the disease was observed in 22 years. DISCUSSION: Aggressive surgery may be indicated in young patients with isolated lesion and good prognostic factors. Palliative surgery remains necessary in order to decompress the cord and to stabilize the spine. PMID- 17121319 TI - Ambulatory proctology surgery--an Indian experience. AB - OBJECTIVE: To evaluate the results of proctological surgeries carried out by us on an outpatient basis. DESIGN: Retrospective study. SUBJECT: 3256 patients with benign ano-rectal pathologies needing surgical intervention. MATERIALS AND METHODS: Patient demographics, type of anesthesia used, the type of operation and postoperative complications are described. Patient satisfaction was assessed independently after surgical procedure. RESULTS: Mean age of the patients was 37.5 years. 61% patients were male. There was no mortality. The mean duration of hospital stay was 8.4 hours (range 3-22 hours). Complication rate was 2.9%. 81.4% patients were highly satisfied with the procedure. CONCLUSION: Day care proctological procedures are a safe and effective way of reducing costs without increasing morbidity, mortality, and is acceptable to majority of patients. PMID- 17121320 TI - Radiofrequency Y-V anoplasty in the treatment of anal stenosis. AB - BACKGROUND: Anal stenosis is a common proctologic disease often caused from hemorrhoids surgery. Treatments adopted are many and varied. In this study we applied radiofrequencies to the classic Y-V anoplasty operation and reported results obtained. METHODS: Eligibility criteria consisted of patients affected by anal stenosis. Exclusion criteria were those with contraindications to the surgical operation: pregnant patients or American Society of Anaesthesiologists Score III or IV. RESULTS: From January 2002 to December 2004 we operated 7 patients, 4 of them were males. Mean age at the time of diagnosis was 46 years. All patients referred obstructive defecation, painful evacuation or bleeding. Mean values for operative time were 30 min. Postoperative pain was 3.9 at day 1 and 3.0 at 7th day (VAS scale). Patient satisfaction rate was 6.6 at three weeks and 8.3 at 6 and 12 months. No recurrences were observed after 1 year. CONCLUSION: Radiofrequency Y-V anoplasty is a feasible and effective for the treatment of anal stenosis. The radiofrequency bistoury easies the procedure, lessens operating times and the healing process of surgical wounds. PMID- 17121321 TI - Amiodarone-induced thyroid dysfunction in clinical practice. AB - Amiodarone is a potent class III anti-arrhythmic drug used in clinical practice for the prophylaxis and treatment of many cardiac rhythm disturbances, ranging from paroxismal atrial fibrillation to life threatening ventricular tachyarrhythmias. Amiodarone often causes changes in thyroid function tests mainly related to the inhibition of 5'-deiodinase activity resulting in a decrease in the generation of T3 from T4 with a consequent increase in rT3 production and a decrease in its clearance. In a group of amiodarone-treated patients there is overt thyroid dysfunction, either amiodarone-induced thyrotoxicosis (AIT) or amiodarone-induced hypothyroidism (AIH). AIT is primarily related to excess iodine-induced thyroid hormone synthesis in an abnormal thyroid gland (type I AIT) or to amiodarone-related destructive thyroiditis (type II AIT). The pathogenesis of AIH is related to a failure to escape from the acute Wolff-Chaikoff effect due to defects in thyroid hormonogenesis, or, in patients with positive thyroid autoantibody test, to concomitant Hashimoto's thyroiditis. Both AIT and AIH may develop either in apparently normal thyroid glands or in glands with preexisting, clinically silent abnormalities. AIT is more common in iodine-deficient regions of the world, whereas AIH is usually seen in iodine sufficient areas. In contrast to AIH, AIT is a difficult condition to diagnose and treat, and discontinuation of amiodarone is usually recommended. In this review we analyse, according to data from current literature, the alterations in thyroid laboratory tests seen in euthyroid patients under treatment with amiodarone and the epidemiology and treatment options available of amiodarone induced thyroid dysfunctions (AIT and AIH). PMID- 17121322 TI - Giant gastric trichobezoar: a direct indication to surgery. PMID- 17121323 TI - [Compared Markov with fractal models by using single-channel experimental and simulation data]. AB - The gating mechanical kinetical of ion channels has been modeled as a Markov process. In these models it is assumed that the channel protein has a small number of discrete conformational states and kinetic rate constants connecting these states are constant, the transition rate constants among the states is independent both of time and of the previous channel activity. It is assumed in Liebovitch's fractal model that the channel exists in an infinite number of energy states, consequently, transitions from one conductance state to another would be governed by a continuum of rate constants. In this paper, a statistical comparison is presented of Markov and fractal models of ion channel gating, the analysis is based on single-channel data from ion channel voltage-dependence K+ single channel of neuron cell and simulation data from three-states Markov model. PMID- 17121324 TI - [Human excited electroencephalogram model and its location in brain]. AB - The aim of this study was to identify the characteristics of human excited electroencephalogram and the excited location in brain. The subjects were excited by taking a fixed quantity of caffeine. Electroencephalographic signals were collected using with 128 channels Phoenix Digital EEG and compared before and after the subjects drank coffee. The results showed obvious differences and compared with the ones before being excited electroencephalogram. After being excited we found that the lower frequency composition was restrained in the region below 10 Hz, the high frequency composition was increased significantly,and a wave crest of 38 Hz was produced in every acquisition point with the height around 200. Then the excited region was located in brain with the software of ASA 3 Course (designed by ANT company of Germany) and found that the excited location was focused on the area of the middle abdomen in the pons' side near bulbar when a person was exciting. PMID- 17121325 TI - [Heart sound recognition algorithm based on PNN for evaluating cardiac contractility change trend]. AB - This paper discusses the recognition of heart sound for evaluating the cardiac contractility change trend, which includes heart sound samples recorded at different exercise condition. Especially, focused on the recognition of heart sound recorded after high intensity exercise workload. The algorithm proposed consisted of two correlative methods. The first was to recognize heart sound recorded at rest and after low intensity exercise workloads by probabilistic neural network and the second was to recognize heart sound recorded after high intensity exercise workloads based on the characteristic of heart sound. Both methods have two consecutive phases. Firstly, all peaks, including the peaks of both heart sounds and noise, are marked by a repetitive threshold detecting algorithm. Secondly, probabilistic neural network is employed to classify the peaks detected in the first phase into Si, S2, and noise. Finally, the performance of the algorithm was evaluated using 45 digital heart sound recordings including normal and abnormal heart sound, which were recorded at rest and after low intensity exercise workloads, and 28 digital heart sound recordings recorded after high intensity exercise workloads. The results showed that over 94% of heart sound samples were classified and recognized correctly. Moreover, the reasons for the wrong classification, of which omitting and misdetection are two main problems, are also discussed and solutions are proposed. So this method can be improved and refined in following studies. In conclusion, this algorithm is a reliable approach to detect and classify heart sounds, providing a solid basis for further heart sound analysis. PMID- 17121326 TI - [The three-dimension medical image reconstruction simulation on 3D Shepp-Logan head phantom]. AB - Shepp-Logan head phantom is the classical model of simulation computing in Computerized Tomography. This paper proposes a new method--considering 3D Shepp Logan head phantom as the basic reference model to carry on simulation experiment and algorithms performance evaluating in three-dimension medical image reconstruction field. The paper introduces the design and the implementation of 3D Shepp-Logan head phantom as well the computing of projection data at first, and then describes the simulation process of the three-dimension medical image. The numerical experiment gives the three-dimension medical image reconstruction simulations based on 3D Shepp- Logan head phantom. The result of this experiment indicates the accuracy of model computing and the feasibility of our this method. PMID- 17121327 TI - [Liquid flow field measurement in a tube with a combination of stenosis and bifurcation]. AB - Two-D liquid velocity field in a tube with a combination of a stenosis and a bifurcation was measured with refractive index matching technique using Laser Doppler Velocimeter (LDV). The flow ratio of main tube to bypass was 1:4. It is found that the axial velocity profile has two peaks in the XOY plane which is skewed to the main tube wall facing the flow from the bypass, and it manifests itself in the form of "M" in the XOZ plane. A two counter-rotating vortices in the main tube cross section downstream the conjunction region is also detected. PMID- 17121328 TI - [Extraction of epileptic spike waves in EEG with B-spline wavelet]. AB - The automatic detection of EEG spikes is important both in diagnosis and alleviating the hart work of doctors. This paper uses B-spline wavelet which does well in detecting break point to detect spike of epilepsy. Through experiment, the validity of the method is proved. PMID- 17121329 TI - [Computer simulation study of the re-entry mechanisms in one-dimensional ischaemic myocardium]. AB - Torsades de Pointes is a kind of severe ventricular arrhythmia. Myocardial ischaemia is one of the major causes leading to TdP. In this paper the mechanisms of the TdP were quantitatively studied under the condition of ischaemia based on the Noble98 dynamic model of the ventricular action potential. The study was conducted on one-dimensional homogeneous myocardium with the method of computer simulation. The models were firstly developed to simulate the lower excitability, extracellular accumulation of the K+ concentration or the decreased gap junctions in ischaemic myocardium. By separately reducing the Na+ conductance, increasing the extracellular K+ concentration or decreasing the conductance of the gap junctions enabled us to study the effect of each change in isolation. Then different degrees of ischaemic models were established to study their physiological features. The study showed that the conduction velocity became slower with the ischaemia aggravation, the action potential duration became shorter and the width of the vulnerable window obviously became larger than the normal conditions. The results illustrated that ischaemia was easily leading to unidirectional conduction block and resulted in re-entry and arrhythmias. PMID- 17121330 TI - [A multi-lead ECG classification network system based on modified LADT]. AB - An electrocardiogram (ECG) classify system based on the features of the ECG and neural network classification, which is the simulation of the real world situation, was present. First, a modified approach of the linear approximation distance thresholding (LADT) algorithm was studied and the features of the ECG were obtained. Then a neural network which can classify the multi-lead ECG data was trained with these features along the theory of the ECG diagnosis and the situation of ECG diagnosis in practice. Thus take a new idea for the ECG automatic analysis. The algorithm was tested using several ECG signals of MIT BIH, and the performance was good. The correct rate of the trained wave is 100%, untrained is 78.2%. PMID- 17121331 TI - [Nonlinear analysis of multi-channel EEG and its application to mental workload detection]. AB - Mental workload research is important to people's health and work efficiency, Psychophysiological measures such as electroencephalography (EEG), ECG and respiration measures can be used to predict mental workload level. A Multi channel phase-space reconstruction method is proposed in this paper which rearranges signal serials by the correlation coefficients and select time delay by signal determinism. The study of determinism and correlation dimension on simulative data exhibits a good performance. The result of EEG series shows a clearly consistency to workload level variety. The method is useful for multi channel signals nonlinear analysis and mental workload detection. PMID- 17121332 TI - [Dynamic analysis of heart rate variability based on wavelet transform]. AB - The analysis of heart rate variability (HRV) has become a tool for noninvasively detecting the cardiovascular modulation of autonomic nervous system. Traditional analysis in frequency domain mainly includes calculating the power and detecting the peak frequency of each physiological frequency band. Whether employing the classical method or AR model to estimate the spectrum, the approximate stationarity of HRV is presupposed. Only in short term analysis can data meet this condition, while in long term the nonstationarity of HRV notably appears. A dynamic analysis method based on wavelet transform was proposed in this paper, which not only can obtain the traditional indices in frequency domain, but can compute their dynamic values varying with time, called short-time power and short time LF/HF ratio. The latter can dynamically evaluate the activity of autonomic nervous. Finally the method was applied to trace the balance of autonomic nervous in Atropin drag experiment. PMID- 17121333 TI - [An improved motion estimation of medical image series via wavelet transform]. AB - The compression of medical image series is very important in telemedicine. The motion estimation plays a key role in the video sequence compression. In this paper, an improved square-diamond search (SDS) algorithm is proposed for the motion estimation of medical image series. The improved SDS algorithm reduces the number of the searched points. This improved SDS algorithm is used in wavelet transformation field to estimate the motion of medical image series. A simulation experiment for digital subtraction angiography (DSA) is made. The experiment results show that the algorithm accuracy is higher than that of other algorithms in the motion estimation of medical image series. PMID- 17121334 TI - [The abstraction of the heart rate signals based on clipped-error LMS algorithm with variable step size of segmenting type]. AB - The adaptive filter is an effective method to dealing with such kind of non steady random signal of the heart rate. The algorithm of clipped- error LMS algorithm with variable step size of segmenting type was proposed. We adopted auto-regression predict filter estimating the noise of heart rate signals through Doppler ultrasound sensor. We used it as the reference signal of the adaptive noise cancelling system, its result is to remove some noise of main input, it made SNR raise by about 36 dB, it realized the abstraction of the heart rate signals. PMID- 17121335 TI - [The application of adaptive algorithm and wavelet transform in the filtering of ECG signal]. AB - Electrocardiographic (ECG) signal are a kind of basic physiological signals of human body, and are very important in clinical diagnosis. But the ECG signals from body surface are often interfered by noises such as 50 Hz noise, baseline displacemant, electromyography (EMG) noise and edv. These noises bring obstacle to the diagnosis of cardiovascular diseases. To eliminate the ECG signals noises mentioned above,this paper adopts LMS adaptive algorithm and wavelet transform theory to design three kinds of digital adaptive filters-adaptive noise cancellation filter, wavelet transform filter and adaptive signal dividing filter to filter the corresponding noises. The results show that the three kinds of noises existing in the ECG signal have been efficiently eliminated. PMID- 17121336 TI - [A method to estimate the short-term fractal dimension of heart rate variability based on wavelet transform]. AB - A new method of calculating fractal dimension of short-term heart rate variability signals is presented. The method is based on wavelet transform and filter banks. The implementation of the method is: First of all we pick-up the fractal component from HRV signals using wavelet transform. Next, we estimate the power spectrum distribution of fractal component using auto-regressive model, and we estimate parameter 7 using the least square method. Finally according to formula D = 2- (gamma-1)/2 estimate fractal dimension of HRV signal. To validate the stability and reliability of the proposed method, using fractional brown movement simulate 24 fractal signals that fractal value is 1.6 to validate, the result shows that the method has stability and reliability. PMID- 17121337 TI - [Wavelet-based pulse-abnormality analysis for heroin addicts]. AB - Using wavelet transforms method, time-frequency characteristics of pulse signals from 15 heroin addicts and 15 healthy persons were analyzed. According to 3-D and contour plots used to display discrete dyadic wavelet transforms, the significant difference of time-frequency characteristics between the signals of heroin addicts and healthy persons were revealed. A primary criterion was also obtained,with the criterion, 15 heroin addicts were entirely identified, while two healthy subjects were misidentified. The research result shows that the wavelet-based multiresolution analysis is a very effective method to extract characteristics of pulse signals. It is valuable to the diagnosis and therapy for heroin addicts. PMID- 17121338 TI - [Research on a novel watershed algorithm combining with wavelet analysis]. AB - Due to noise and irregularity of the gradient image, watershed algorithm used to segment images generally leads to over-segmentation, which is unacceptable. A novel watershed algorithm, which combines with some concepts of wavelet analysis and mathematic morphology, is proposed in this paper. The test shows that this method can solve the problem of over-segmentation and achieve the anticipative goal. PMID- 17121339 TI - [Visualization and real-time tracking of the instruments in surgical navigation system]. AB - In order to realize the visualization and real-time tracking of the instruments in surgical navigation system, we use the CAD solid model of the instruments directly or by reverse engineering, and transform the solid model to surface model. Then, the instruments, which are fixed with locator, are characterized and tested their precision. After registration between the CT model and the patient site in intra-operation, the system accurately displays the STL model of the instruments according to the tracked site and orientations. The model construction of the instruments is the base of the surgical navigation and is important to improve the security and precision of operation. PMID- 17121340 TI - [A study on clustering analysis of arrhythmias]. AB - According to the characteristics of ECG analysis, large data quantum, high accuracy demand and real-time, a classified algorithm of arrhythmia based on clustering analysis is presented in this paper. According to "things-of-one-kind come-together" principle, this algorithm uses the similarities of cases with same kind of heart disease at the same time, includes the factors of the individual difference to analyze arrhythmias by clustering QRS complex waveform and rhythm analysis as the subordinate method. Verified by eight records of MIT-BIR standard heart electricity database, the probability of correct clustering reaches above 90%, which shows that this algorithm can analyze arrhythmias effectively. PMID- 17121341 TI - [An improved fast algorithm for ray casting volume rendering of medical images]. AB - Ray casting algorithm can obtain better quality images in volume rendering, however, it presents some problems such as powerful computing capacity and slow rendering velocity. Therefore, a new fast algorithm of ray casting volume rendering is proposed in this paper. This algorithm reduces matrix computation by the matrix transformation characteristics of re-sampling points in two coordinate system, so re-sampled computational process is accelerated. By extending the Bresenham algorithm to three dimension and utilizing boundary box technique, this algorithm avoids the sampling in empty voxel and greatly improves the efficiency of ray casting. The experiment results show that the improved acceleration algorithm can produce the required quality images, at the same time reduces the total operations remarkably, and speeds up the volume rendering. PMID- 17121342 TI - [Finite element analysis of the mechanical property of the resistance to compressing of the coronary stent]. AB - Since coronary stent is an important device used in percutaneous support the stenosed artery, its ability to expant the diseased artery wall shall be one of the most significant prerequisites for optimal stent performance. In this paper, the finite element model of coronary stent to resist compressing was set up based on the stent prototype testing. Using the numerical simulation, it was systematically analyzed that the influence of the size of the dilatation of stent and stent strut width and thickness on the stent mechanical properties of the resistance to compressive force applied by two opposite planes. The results show that this resistibility is weakened with the increasing of the final expanded diameter of stent, but increasing the stent strut width or thickness can improve it, and it is equivalent to this improvement to increase the dimension of strut cross section of the stent in the two directions. Experiment result is well consistent with simulation which indicates that the finite element method can substitute for the stent prototype testing to some extent. PMID- 17121343 TI - [Experimental study on the migration of vascular endothelial cells stimulated by IL-8]. AB - To investigate the influence of different concentrations of IL-8 on the migration of vascular endothelial cells and find out the best IL-8 concentration, the transwell chamber motility assay and the scrape motility assay were applied to observe the migration of vascular endothelial cells induced by IL-8. The results demonstrated that the migration of vascular endothelial cells was increased significantly under different IL-8 concentrations, while the best effect occurred when IL-8 concentration was 100 ng/ml. PMID- 17121344 TI - [Study on the biomechanical behavior of human intestine]. AB - To investigate the biomechanical behavior of human intestines. The tensile test human intestine was performed with the electronic tension machine in this paper. The results indicate that the exponential relationship for the stress-strain of the human intestine was obtained, and the exponential coefficient a of each segment of the intestine is almost the same although the constant C is different. It also shows that the relative rate of stretch length of each segment intestines is different in longitudinal and circumferential directions. And the incremental elastic modulus of colon is less than those of small intestine. It is considered that the colon can be more easily deformed. The experimental results provide the theoretic basis for research on intestinal endoscopic microrobot. PMID- 17121345 TI - [The effect of aerobic and anaerobic endurance training on the regulating function of autonomic nervous system and its significance]. AB - To evaluate the effect of aerobic and anaerobic endurance training the regulating the function of autonomic nervous system, in order to provide scientific basis for optimizing the project of physical fitness training. Fourty-one healthy young men were randomly divided into aerobic and anaerobic endurance training groups. The training period was 8 weeks. Pre-exercise, 4 weeks and 8 weeks after trained, HRV were measured compared with pre-exercise. The autonomic balance in aerobic endurance group had an increasing parasympathetic activity (HF, HFnu, RMSSD, PNN50, all P was < 0.05) and relatively decreasing sympathetic activity (LFnu). This group showed a parasympathetic predominance (LF/HF) and increase of HRV. While in the anaerobic group there was a relative stabilization with the function of autonomic nervous system. The present study shows that the effect of aerobic and anaerobic endurance training on the autonomic nervous system depends on its intensity. Proper intensity of anaerobic endurance training may be beneficial to improve the adaptability of human body for circumstances as aerobic endurance training. PMID- 17121346 TI - [Effects on biomechanical properties of hip articular cartilage degeneration following the femoral neck fracture in rabbits]. AB - Osteotomies were performed through post-lateral approach under the base of femoral neck on 60 New Zealand white rabbits. The changes of the thickness and biomechanical properties of acetabular cartilage following the cartilage degeneration and the relationship to the postoperative time 2, 4, 6, 8, 12, 16 weeks later were measured respectively. The cartilage specimens were harvested and measured by the automated creep indentation apparatus to obtain the thickness and biomechanical properties. The cartilage became thicker within 6 weeks after operation, cartilage degeneration developed critically from the surface with the lost of the biomechanical properties. The degeneration progressed till the eartilage became thinner and thinner and the to be torn out into pieces at last. The matrix of articular cartilage degraded markedly for the absence of normal stress load following the femoral neck fracture, led to the decline of the biomechanical properties and kept the degeneration progress to osteoarthritis. PMID- 17121347 TI - [Finite element analysis (FEA) for the structure capacity of proximal femur during falling--(I) FEA model and the failure criteria for the bone]. AB - Femur fracture from falls is considered one of the most serious types of the elderly. FEA has proved to be an extremely useful tool in the structure analysis of the proximal femur. In this paper, a FEA model of proximal femur is introduced, and Hoffman failure criteria are built based on the experimental strength data for both cortical bone and trabecular bone of the femur from some references. The FEA model and the failure criteria are verified using other researcher's experimental results. The predicted trabecular failure load was only 0.5% lower than the experimental data and cortical failure load was 4.2% higher than the experimental result. This result shows that our FEA model, combined with the Hoffman criterion for both cortical bone and trabecular bone, can effectively predict the structural capacity of the femur during falling. PMID- 17121348 TI - [Investigation of computational fluid dynamics application in blood pumps]. AB - One of the internal reasons resulting in hemolysis and thrombi is the hemodynamics. Many studies show that irregular flow patterns and shear stress result in the damage of blood cells. With the rapid development of computer technology, simulation for such microdynamics becomes possible. Computational fluid dynamics was applied to predict the flow in the streamlined blood pump and a pump with straight vanes. After the analysis of flow patterns and the distribution of shear stress, it was concluded that in the same boundary conditions, the blood pump based on streamlined design had better hemodynamics than the pump with straight vanes and caused less PMID- 17121349 TI - [Strain analyses of left ventricle wall with spatial modulation of magnetization based on two-dimensional method]. AB - We proposed a two-dimensional deformation analysis method in regional ventricular wall motion by using tagged magnetic resonance (MR) images to evaluate the wall functions. In a region surrounded by four image slices, a material point is traced to obtain the displacement and strains. MR images were taken 24 phases over one cardiac cycle in the long and short axes of heart. PMID- 17121350 TI - [The biomechanical study of rotating-arm self-locking intramedullary nails in comminuted femoral shaft fractures]. AB - This study was sought to compare and evaluate the biomechanical properities of rotating-arm self-locking intramedullary nails (RSIN) with domestic femoral intramedullary nail (DFIN) in communited femoral shaft fractures. A 2 cm midshaft defect was created on seven pairs of cadaver femur, and fixed with two kinds of intramedullary nails. Then the torsion, four-point bending and axial loading to failure were tested on these models. The load versus deformation curve was generated from the load recorded by the load cell and the deformation simultaneously recorded by the linear variable displacement transducer. The mean torsional stiffness of the femora fixed with RSIN was 0.98 Nm/degree, which was significantly less than that fixed with the similar nails reported in documents, and the mean value of bending stiffness of the femur fixed with DFIN was larger than that of the femur fixed with RBIN, but there was no significant statistic difference. For use in fracture models under compression load beyond 1200 N, the RBIN was more rigid than DFIN, which provided the biggest load. All the parameters of the test with nails were greater than that for femur fixed with nails. In conclusion, FBIN probably provides enough strength and rigidity for use in communited femoral shaft fractures. PMID- 17121351 TI - [An association study between the insertion/deletion polymorphism of angiotensin I converting enzyme gene and human speed endurance]. AB - This study was conducted to research the association between the insertion/deletion (I/D) polymorphism of Angiotensin I converting enzyme (ACE) gene and human speed endurance. Fourty subjects of Han nationality, healthy, with similar sports history were included. The I/D polymorphism of ACE gene was detected by polymerase chain reaction (PCR). The score of 800m run and the concentrations of the whole blood lactic acid were mensurated. Cluster analysis of the grade was made according to the result of cluster analysis. The subjects were divided into two groups: high speed endurance group and low speed endurance group. We found that both the distributions of the ACE genotypes and the distributions of the ACE alleles there were no significant difference between high speed endurance group and low speed endurance group (P > 0.05); Whether at rest state, or after 800m run or the difference value between rest and after 800m run,the concentrations of the whole blood lactic acid did not exist significant difference among three kinds of genotypes groups (P > 0.05). There was on association with I/D polymorphism of ACE gene and human speed endurance. PMID- 17121352 TI - [Preparation and properties of medical calcium phosphate cement]. AB - The preparation of tetracalcium phosphate (Ca4(PO4)2O, TTCP)was studied. Then calcium phosphate cement (CPC) was prepared. The setting time, pH value, compressive strength, X-ray diffraction (XRD) and scanning electron microscope (SEM) analysis of CPC were studied. The results show that TTCP containing small amount of CaO can be successfully attained heated at 1 500 degrees C for 6 h in vacuum condition. The initial setting time and final setting time of CPC is 4 min and 15 min, respectively. Its compressive strength is 20 MPa after 1-day immersion and 35 MPa after 7-day immersion. The pH value of the solution changes between 6.4 and 8.9. These properties can satisfy the clinical requirements of CPC. The final product of CPC is flake-like or needle-like hydroxyapatite (Ca5(P04)3OH, HA). The continuous network structure of HA appears in the microstructure, this leads to increase the strength of the material. This CPC can be used as bone substitute material. PMID- 17121353 TI - [Analysis of FT-IR-ATR spectra of serum proteins adsorbed on carbonaceous materials]. AB - To clarify the reason causing difference of serum proteins adsorbability on different carbonaceous materials, FT-IR-ATR spectra of human serum albumin (HSA) and human serum fibrinogen(HFG) before and after adsorbing on diamond like carbon film (DLC),diamond film (DF) and graphite were analyzed. It has been shown that there are hydrogen bond because of -NH at the interfaces of HSA-DLC, HFG-DF and HFG-graphite. Based on the results, earlier research conclusion that the adsorbability of HSA on DLC higher than that on DF and graphite, but on DF and graphite the adsorption of HFG takes precedence can be explained rationally. PMID- 17121354 TI - [Preparation of graded zirconia-hydroxyapatite composite bioceramic and its immunocompatibility in vitro]. AB - Graded Zirconia-hydroxyapatite composite bioceramic and simplex Zirconia hydroxyapatite composite bioceramic were produced, and the extractes of these two of materials were made to evaluate their immunocompatibility. Scanning electron microscope (SEM) was used to detect the character of the surface of the graded composite bioceramic. Holding and without holding phytohemagglutinin (PHA), proliferation and activation of peripheralblood monocytes(PBMCs) cultured in the two extracts were studied. Cultured in PHA after 72 hours, the proliforation rate of the graded composite group was significantly higher than the simplex composite group (P < 0.01). There was no difference of apoptosis of PBMCs of the two groups (P > 0.05). Cultured in PHA after 24 hours, the ratio of CD3/CD69 positive PBMCs of the simple composite group was significantly higher than that of the graded composite material group (P < 0.01). The numbers of PBMCs activated by the graded composite material group were less than that of the simply composite material group and the technique of graded composite will be helpful to improve its immunocompatibility. PMID- 17121355 TI - [Effect of degradable products on degradable property of PGLA in vitro]. AB - In order to investigate the effect of degradable products on degradable property of PGLA in vitro, two kinds of media-PBS and artificial plasma were prepared for immersing PGLA under changing or non-changing media condition. The mass loss rate of PLGA was calculated and the pH value in the non-changing media was measured before and after immersing 2 w, 3 w, 4 w, 6 w, 8 w and 10 w respectively. The results showed that there was almost no statistically significant difference of mass loss rate of PGLA immersing in two kinds of media at 2 w (P > 0.05). But from 2 w to 6 w, the degradation of PGLA immersing in both media under non changing media group was remarkably faster than those at the same period of changing media group (P < 0.01). During the whole degradable period, the pH value in PBS kept stable around 7.0-7.4, while the pH value in artificial plasma showed gradually decreased as the degradation of PGLA from 7.5 to 5.7. The change of pH values had statistically significant difference between two degradable media (P < 0.01). It was implied that the degradable products existed in immersing media had an effect on degradable speed of PGLA itself if the media was unchanged. It could accelerate the mass loss of material. The pH value also affected the degradable property of PGLA, the lower the pH value, the slower the degradable speed. PMID- 17121356 TI - [Preparation of cytarabine nanoparticle and study on the drug releasing mechanism]. AB - Cytarabine nanoparticle was prepared through emulsion polymerization method, and its releasing properties were studied. The results showed that releasing principle complied with biexponential equation and had characteristics of sustained releasing. The pharmacokinetics in rabbits complied with two-department model. Compared with cytarabine injection, cytarabine nanoparticle had prolonged t 1/2beta and MRT and reduced CL, which showed nanoparticle could significantly prolong the retention time of Ara-C and possess significant sustained releasing character. PMID- 17121357 TI - [Encapsulating hepatocytes with chitosan in physiological conditions]. AB - Prepared from 15.3% N-acetylated chitosan (FNC), half N-acetylated chitosan (HNC) possesses a good solubility in a weak basic solution, guaranteeing the formation of microcapsules by the coacervating reaction between HNC and methacrylic acid (MAA)-hydroxyethyl methacrylate (HEMA)-methyl methacrylate (MMA) (MAA-HEMA-MMA) terpolymer under physiological conditions. When hepatocytes were encapsulated in such 3-dimensional microenvironment, as compared to monolayer culture, cell functions, including P450 activity, urea production and albumin release, were well supported. The prepared microcapsules have good mechanical stability and permeability. PMID- 17121358 TI - [Sol-gel preparation of ultrathin nano-hydroxyapatite coating and its characterization]. AB - Present study used dip-coating techniques to fabricate ultrathin nano-HA coating on titanium in organic sol-gel of Ca (NO3)2. 4H2O and PO(CH3)3 and inorganic sol gel of Ca (NO3)2. 4H2O and (NH4)2HPO4. Scanning electron microscope (SEM) and grazing-incidence X-ray diffraction (XRD) were used to observe the morphology and distribution of crystallite size (D) and lattice strain (epsilon) of ultrathin nano-HA coating. After heated at 400 degrees C, the apatite structure of coatings on titanium began to appear. At heating temperature of 400 degrees C-600 degrees C, the effect of heating temperature on D and epsilon of both coatings was obvious. Precursor types significantly affected the particle diameters of nano-HA coatings, which were 25-40 nm for organic sol-gel and about 100 nm for inorganic sol. The thickness of ultrathin nano-HA coatings was 2.5 microm for organic sol gel and 5 microm for inorganic sol and morphology of interfaces between coating and titanium was intact and homogenous. PMID- 17121359 TI - [Rhythmic beating cardiomyocytes derived from human embryonic germ (EG) cells in vitro]. AB - Embryonic germ (EG) cells are pluripotent cells derived from primordial germ cells (PGCs) of gonads, gonadal ridges and mesenteries, analogies of fetuses,with the ability to undergo both highly self-renewal and multiple differentiation. These cells in vitro can differentiate into derivatives of all three embryonic germ layers when transferred to an in vitro environment and have the ability to form any fully differentiated cells of the body. The aim of this study is to investigate the potentiality of human EG cells differentiation into cardiomyocytes. Inducing human EG cells with the method of murine ES cells differentiation into cardiomyocytes, supplemented with 0.75%-1% DMSO, 20% NBS, 10(-7) mM RA and 20% cardiomyocytes conditioned medium. RESULTS: 20 heart-like (rhythmic beating cell masses were observed in vitro culture and delayed human EG cells, which beat spontaneously from 20-120 times per minute and maintained beating for 2-15 days, periodic acid's staining (PAS), Myoglobin and a-actin immunological histology positive were all positive and reacted with K+, Ca2+ and adrenalin. Relatively unorganized myofibrillar bundles or more organized sarcomeres, z-bands or a gap junction, the presence of desmosomes in a few cells of the cell masses was observed with transmision electron microscope, which initially demonstrated that these cells were cardiomyocytes. We could not get rhythmly beating cardiomyocytes with 0.75%-1% DMSO, 10-7 mM RA and 20% cardiomyocytes conditioned medium,but in which the percentage of cardiac alpha actin immunostaining positive cells were increased. The results first demonstrated that human EG cells can differentiate into rhythmic beating cardiomyocytes in vitro and suggests that human EG cells may represent a new potent resource for cardiomyocytes transplantation therapy for myocardium infarction. PMID- 17121360 TI - [Fusion expression and identification of angiostatin and endostatin in E. coli BL21(DE3)]. AB - Angiostatin(AS) and endostatin(ES) are both potent endogenous angiogenesis inhibitors, and the combination of AS and ES has been shown to have synergistic antiangiogenic effects. Here we report the fusion protein AS-ES expressed in E. coli which has antiangiogenic effects. At first, AS and ES genes were cloned respectively through RT-PCR, then fusion gene was made through gene splicing ,finally pET-42 (b)/AS-ES expression plasmid was constructed and transduced in E. coli BL21 (DE3). Target protein was in form of inclusion body,the rate of expression was about 14%, and MW about 65KD. Western blotting assay showed expressed protein had specific immune reaction to both the antibodies of AS and ES. The expressed protein which was refolded and purified through heparin affinity chromatography had antiangiogenic effect to vessels on chicken embryo chorioallantoic membrane. The results show that fusion protein AS-ES was expressed successfully in E. coli, and the expressed protein,which was renatured and purified, had immuno-reactivity to anti-AS and anti-ES in Western blotting and angiogenesis inhibition activity. PMID- 17121361 TI - [Transfection of dominant negative MyD88 decreases IL-8 production in bacteria infected airway epithelial cells]. AB - Interleukin-8 (IL-8) is an important activator and chemoattratant of neutrophils and has been implicated in airway inflammatory diseases. To explore the new gene therapeutic strategies for airway inflammation, plasmid expressing dominant negative myeloid differentiation protein (MyD88 DN) was constructed and transfected into human airway epithelial cell lines A549 and SPC-A-I. The cells were challenged with M. tuberculosis, P. aeruginosa or K. pneumoniae and the release of IL-8 was measured using ELISA. The results showed that the supernatants of M. tuberculosis and R. aeruginosa enhanced IL-8 release from the epithelial cells; and transfection of MyD88 DN diminished this effect. MyD88 DN also reduced IL-8 release from cells induced by live bacteria of P. aeruginosa or K. pneumoniae. These data suggest that MyD88 could be used as a target gene in the gene therapy of airway inflammation. PMID- 17121362 TI - [Inhibition of cell proliferation and C-myc cancer protein expression in human colon adenocarcinoma cell line HT29 with VIP-131I-ASON]. AB - A 15-mer phosphorothioate antisense oligonucleotide (ASON) complementary to the translation start region of the C-myc oncogene mRNA was labeled with 131I or 125I and the labelled compound was linked to the vasoactive intestinal peptide (VIP) to be bound covalently to a polylysine chain so as to deliver oligonucleotide into tumor cells. The effect of the VIP as carrier on cell uptake of ASON in tissue culture was evaluated in a human colon adenocarcinoma HT29 cell line. The efficacy of VIP-131-ASON on cell growth was evaluated using the MTT assay. Expression of c-myc-encoded protein was measured by flow cytometry. Sense and nosense control Oligonucleotides with VIP carrier were used as control. The results showed that VIP competed effectively with VIP-125I-ASON to bind the HT29 cells. Cell uptake was increased 3-4 fold using the VIP carrier compared to the same dosage of naked DNA. HT29 cells treated with VIP-131I-ASON complexes exhibited 4-fold lower proliferation than those treated with 13I-ASON and 6-fold lower proliferation than those treated with radioiodinated Sense and nosense DNA. Cancer protein expression of HT29 cells treated with VIP-131I-ASON was decreased 2-fold compare with that in 131I-ASON treated cell. The use of a VIP carrier greatly increased 131I-ASON cellular uptake and inhibition of cell proliferation and C-myc cancer protein expressing in HT29 cell by radioiodinated antisense Oligonucleotides. PMID- 17121363 TI - [Study of a new medical stainless steel]. AB - Medical implantable stainless steels are widely used in medical field due to their excellent properties, besides its allergic response to human body, the nickel ion released from the steels due to corrosion has the harm of malformation and carcingenesis. The mechanical property, corrosion resistance and blood compatibility of a new nickel-free stainless steel (BIOSSN4) is researched in this paper. Compared with the traditional 316L medical stainless steel, BIOSSN4 shows wide future applications because of its better combination of strength and toughness, good corrosion resistance and biocompatibility. PMID- 17121364 TI - [The dynamic concision for three-dimensional reconstruction of human organ built with virtual reality modeling language (VRML)]. AB - This research studies the process of dynamic concision and 3D reconstruction from medical body data using VRML and JavaScript language, focuses on how to realize the dynamic concision of 3D medical model built with VRML. The 2D medical digital images firstly are modified and manipulated by 2D image software. Then, based on these images, 3D mould is built with VRML and JavaScript language. After programming in JavaScript to control 3D model, the function of dynamic concision realized by Script node and sensor node in VRML. The 3D reconstruction and concision of body internal organs can be formed in high quality near to those got in traditional methods. By this way, with the function of dynamic concision, VRML browser can offer better windows of man-computer interaction in real time environment than before. 3D reconstruction and dynamic concision with VRML can be used to meet the requirement for the medical observation of 3D reconstruction and has a promising prospect in the fields of medical image. PMID- 17121365 TI - [Prediction of protein solvent accessibility with Markov chain model]. AB - Residues in protein sequences can be classified into two (exposed / buried) or three (exposed/intermediate/buried) states according to their relative solvent accessibility. Markov chain model (MCM) had been adopted for statistical modeling and prediction. Different orders of MCM and classification thresholds were explored to find the best parameters. Prediction results for two different data sets and different cut-off thresholds were evaluated and compared with some existing methods, such as neural network, information theory and support vector machine. The best prediction accuracies achieved by the MCM method were 78.9% for the two-state prediction problem and 67.7% for the three-state prediction problem, respectively. A comprehensive comparison for all these results shows that the prediction accuracy and the correlative coefficient of the MCM method are better than or comparable to those obtained by the other prediction methods. At the same time, the advantage of this method is the lower computation complexity and better time-consuming performance. PMID- 17121366 TI - [Quadrature Doppler ultrasound signal denoising based on adapted local cosine transform]. AB - The spectrogram of Doppler ultrasound signal has been widely used in clinical diagnosis. The additional frequency components arising from internal or external noise to the system will produce adverse effects on its subjective and quantitative analysis. A novel approach based on the adapted local cosine transform and the non-negative Garrote thresholding method was proposed to remove noise from quadrature Doppler signal. At first, the directional information was extracted from the quadrature signal. And then the denoising method based on the adapted local cosine transform is performed on the forward and backward flow signals, respectively. At last, the estimated signal was reconstructed from the denoised signals using Hilbert transform. In the simulation study, both the mean frequency and spectral width waveform were studied for the denoised signal. The simulation results had shown that this approach was superior to that based on the wavelet transform, especially under low SNR conditions. PMID- 17121367 TI - [Adaptive exercise electrocardiographic signal enhancer with manual neural network anticipate filtering ]. AB - Exercise electrocardiogram is to detect ECG signals when one is in exercise state. Its characteristics are that the exercise will result in serious excursion of the baseline ,the distinct increase of the muscle voltage interfere and the fall signal-to-noise. This article introduces a adaptive exercise ECG signal enhancer with manual neural network. It is designed by combining non-linearity of manual neural network and the tracking characteristics of adaptive processing. It can reduce the noise, increase signal-to-noise and effectively abstract exercise PMID- 17121368 TI - [Review of nonlinear filters for medical ultrasonic images]. AB - Ultrasound medical imaging has been widely applied in clinic diagnoses because of its real-time, non-invasiveness and convenience. However, it suffers from severe speckle noises. Until now, a lot of filtering algorithms have been proposed, but none of them is satisfactory. In this paper, four nonlinear filter methods, based on median filter, wavelet decomposition and anisotropic diffusion, are systematically reviewed. Finally, the prospect of the nonlinear filtering method is discussed. PMID- 17121369 TI - [Applications of polymerase chain reaction biochip/microdevice technology in the clinical diagnosis]. AB - The polymerase chain reaction (PCR) technique has been widely used in fields of molecular biology and diagnosis. PCR biochip/microdevice is increasingly of great interest as a result of its small volume of sample and reaction mixture, short reaction period and portability. The applications of PCR biochip/microdevice are specially introduced in the clinical diagnosis. Finally, the applications and development of PCR biochip/microdevice are also predicted. PMID- 17121370 TI - [Design of a cardiac monitoring system using mobile communications]. AB - Telemedicine is gaining more and more momentum as a new approach for patients'surveillance. In many monitoring parameters, electrocardiography is the most important, especially for monitoring patients with serious cardiovascular diseases. They need to be monitored frequently in order to detect any deterioration. In this paper,we design a electrocardiographic telemetry system using General Packet Radio Service(GPRS). This design adds to traditional Holter capability of monitoring the patients'heart anywhere, anytime. PMID- 17121371 TI - [Gene chip application in organ transplantation and ischemia reperfusion injury]. AB - Gene chip has been an important way to detect gene changes of organism in different condition. It is a new tendency to use gene chip to research graft rejection at gene level. Ischemia reperfusion injury (IRI) accurs early in organ transplantation. Studying IRI with gene chip is beneficial to understand the mechanism of graft rejection and will provide guidance for surveying and treating graft rejection after organ transplantation. PMID- 17121372 TI - [Comparison of the clinic results of speech processing strategies and latest advances on cochlear implant]. AB - Cochlear implant (CI) is a neural prosthetic device used to provide the sensation of sound to those who are profoundly deaf by delivering electrical stimulus to auditory nerve directly . It is becoming one of the main research frontiers in the area of otology and rehabilitation engineering . This symposium proposes an introduction about the new achievements of cochlear implant research , which consists of the following parts: the structure of CI, new speech processing strategies, comparison of the clinical results of speech processing strategies and the hotspots of the research. PMID- 17121373 TI - [Alzheimer's disease and oxidative stress]. AB - Alzheimer's disease (AD) has abstracted many scientists' interests with the aging of people in the world, but its mechanisms are still remaining unclear. This review will focus on some of the mediators of oxidative stress occurring in AD pathology and their possible role in the AD pathogenesis. Meanwhile, antioxidant approaches for the prevention and treatment of AD are discussed. PMID- 17121374 TI - [A new real-time method monitoring behaviors of living cells in vitro-PCC]. AB - A method combinating piezoelectric-biochip technique with cell culture technique in vitro has been developed. Piezoelectric cell-based chip (PCC) can continously monitor the behaviors of living cells attached to or spreaded on surfaces. The method measures the resonant frequency (f), the dissipation energy (D) and impedance (Z), in real time. These multi parameters can inflect information of cells qualitively and quantitively. This paper covers principle, materials and methods, results and discussion of the new technique, simultaniously indicates its application and trends. PMID- 17121375 TI - [Think about it!]. PMID- 17121376 TI - [Significance of color Doppler ultrasonography in the assessment of pancreatic carcinoma vascular invasion]. AB - BACKGROUND/AIM: It is highly appreciated to provide exact data on vascular invasion of pancreatic carcinoma relying as much as possible on non-invasive diagnostic procedures. Color Doppler ultrasonography has been proven as an efficient method for clinical staging of pancreatic carcinoma essential for therapeutic decisions. The aim of this study was to provide an analysis of the sensitivity and specificity for color Doppler ultrasonography in patients suffering from pancreatic carcinoma. METHODS: We performed color Doppler ultrasonography examination in 43 patients with pancreatic carcinoma prior to the surgery. The findings of ultrasonography on neoplasm vascular invasion were correlated to the findings obtained during the subsequent surgical procedures. An estimation of neoplastic invasion of certain blood vessels including portal vein, celiac trunk, and superior mesenteric artery and vein is critical for decision making regarding surgical treatment. The patients with metastases of pancreatic carcinoma were excluded from the study. RESULTS: Comparing color Doppler and the surgical findings we estimated the sensitivity for detection of neoplastic vascular invasion ranging from 79-93%, whereas the specificity range was from 83 93%. CONCLUSION: Color Doppler ultrasonography is a sufficiently sensitive and specific method for evaluation of vascular invasion in pancreatic carcinoma patients. Since color Doppler ultrasonography is a non-invasive, radiation free, and inexpensive diagnostic tool, considering also the results of this and similar studies we could strongly recommend its use for an initial presurgical evaluation of vascular invasion in pancreatic carcinoma patients. PMID- 17121377 TI - [Clinical characteristics of multiple system atrophy in serbian population]. AB - BACKGROUND/AIM: Mulstiple system atrophy (MSA) is a neurodegenerative central nervous system disorder, characterized by any combination of extrapyramidal, cerebellar, pyramidal or autonomic disturbance. The aims of our study were to define clinical characteristics of MSA patients in our population, to account for neuroradiological and electrophysiological profile of the disease and to evaluate one-dose levodopa response. METHODS: We have diagnosed 29 patients as MSA, with disease duration from the first symptom 5 years on average on examination. The examinating procedure included an anamnesis and complete neurological investigations, as well as neurootological and neuroradiological examinations. The study included the patients of the Institute of Neurology of the Clinical Center of Serbia in the period of 1996-2001, who completed both clinical and diagnostic criteria for a possible and probable MSA. RESULTS: Autonomic disturbances were documented in 93.1%, whereas extrapyramidal symptoms were seen in additional 89.3% with symmetrical onset in 60%. Levodopa response was poor or moderate. Cerebellar signs were present in 63%, while pyramidal signs occured in 78.7%. There was no a cognitive deterioration (MMSE > 24). CT scan and MRI showed cerebellar and brainstem atrophy, as well as diffuse cortical atrophy. CONCLUSION: Failure of additional diagnostic procedures to distinguish MSA patients required a precise understanding of their clinical specificities. Our results support this statement. PMID- 17121378 TI - [Mobility of B.C.T. dental implants inserted by one- and two-phase surgical method--an experimental study]. AB - BACKGROUND/AIM: Achievement of the osseointegration of dental implants is of crucial importance for their long-term survival. One of the factors that influence the osseointegration is a surgical method of implantation. The outcome of osseointegration can be evaluated on the basis of implant mobility in bone. The aim of this study was to investigate and compare the mobility of B.C.T. dental titanium implants inserted to experimental animals using an one- and two phase method. METHODS: The investigation was performed using a split-mouth design on nine dogs, male german shephards, average age of 3.5 years and average weight of 32 kg. Extractions of the third and fourth lower premolars were performed under intravenous (i.v.) anaesthesia with 5% ketamine chloride. Eight weeks after the extractions, the implants, diameter of 4.5 mm each, with four threads 13.7 mm long, were inserted. Eighteen implants were inserted one side of the jaw using a one-phase method, and another 18 implants were inserted contralaterally using a two-phase method. Three months after the implantation, the implant mobility was evaluated. Three measurements were performed with a Periotest device, and average values were calculated. The implant mobility was classified according to the Periotest scale in four groups of Periotest values (PTV) and compared. A total of 36 implants were inserted in 9 experimental dogs. The PTV ranged from -7.666 to + 50. RESULTS: According to the Periotest scale, 14 one-phase implants (78%) were classified into the 0 group of PTV, and 4 one-phase implants (22%) in the 3rd group. Thirteen two-phase implants (72%) were classified in the 0 group, and 5 implants in the 3rd group of Periotest scale. The difference in the average values of PTV between the two methods was 0.879 which was not statistically significant. CONCLUSION: This study showed that the method of implant insertion had no influence on the implant mobility, i.e. satisfactory osseointegration could be achieved by both methods. Further clinical parameters, as well as pathohistological and histomorphometric ones, have to be evaluated in order to assess better outcome of a particular method. PMID- 17121379 TI - [Preparation and testing of buffered eye drops containing pilocarpine chloride with timolol maleate]. AB - BACKGROUND/AIM: In glaucoma therapy beta-blocator, timolol-maleate, is the first line medicine of selection, whereas a miotic, pilocarpine chloride, is one of the oldest medicines used for the treatment of this illness. They are applied as eye drops. In order to achieve a better therapeutic effect and improve life quality of the ill, we produced and tested eye drops formulations based on combining pilocarpinechloride and timolol maleate in buffers of diferent pH values. METHODS: Following the general pharmacopoeial eye drops preparation regulation, we prepared formulations, of solution type, of pilocarpine chloride and timolol maleate combination, of pilocarpine choloride alone and of timolol maleate alone. A modified phosphate buffer according to Sorensen at 7.4, 7.7 and 8.0 pH values was used as a solvent. The quality of the produced formulations was examined using physical and physico-chemical methods and biological tests. Following pharmacopoeial regulations, we examined clarity, pH value and sterilty. Pilocarpine chloride level was determined by means of ion-par (High Performance Liguid Chromatography) RP-HPLC method, whereas UV/VIS absorption spectrophotometry was used for determining the level of both timolol maleate, and timolol. RESULTS: Results showed that monocomponent and combined preparations complied with the regulation demands. With the increase in the pH value of the solution pilocarpine chloride level decreased in relation to its initial content, whereas timolol level showed a tendency of moderate increase. CONCLUSION: Magistrally prepared pilocarpine chloride eye drops with timolol maleate have satisfied all the required conditions for an ophtalmological preparation. A modified phosphate buffer according to Sorensen at 7.4 pH value proved to be the most optimal solvent. PMID- 17121380 TI - Therapeutic value of serum uric acid levels increasing in the treatment of multiple sclerosis. AB - BACKGROUND/AIM: Uric acid was successfully used in both, prevention and treatment of the animal model of multiple sclerosis (MS). Recently it has been shown that inosine, a ribosylated precursor of uric acid, might be used to elevate serum uric acid levels in MS patients. The aim of this study was to evaluate the safety and efficacy of oral inosine as a single drug treatment in patients with MS. METHOD: We administered inosine orally to 32 MS patients from 2001-2004 year at doses from 1-2 g daily (given twice) depending on the pretreatment serum uric acid levels. The mean follow-up interval was 37.69+/-6.55 months. The other 32 MS patients, without any treatment except for a relapse period (matched by age, sex, duration of disease and functional disability), were used as controls. The follow up interval of these patients was 36.39 +/- 2.68 months. The neurological disability was evaluated by the Expanded Disability Status Scale score (EDSS). RESULTS: During the observed period the treated MS patients were found to have the lower relapses rate than the non-treated MS patients (Chi-square test, p = 0.001). None of the patients have showed any adverse effect of inosine treatment. The non-treated MS patients were found to have a higher increasing in the mean EDSS score than the treated ones (two-way ANOVA-repeated measures/factor times, p = 0.025). CONCLUSION: Our results suggested that the treatment approaches based on the elevation of serum uric acid levels might prove beneficial for some MS patients. PMID- 17121381 TI - Psychosocial and genetic determinants of tobacco smoking as an addictive disease. PMID- 17121382 TI - The effect of tobacco smoke ingredients on immunity with special reference to chronic obstructive pulmonary disease. PMID- 17121383 TI - Influence of tobacco smoke on the appearance of oxidative stress in patients with lung cancer and chronic obstructive pulmonary diseases. PMID- 17121384 TI - [Supraselective embolization as a method for the management of renal vascular injuries]. AB - BACKGROUND: Renal injuries are most often caused by traumatic injuries, but they can also be induced iatrogenically, during renal biopsy, percutaneous nephrostomy or lithotripsy. Vascular renal injuries may be treated by embolization, non surgical or surgical methods. CASE REPORT: In this paper we presented a high-risk patient with poor outcome of an open surgery threatment and a prior attempt of embolization, with gross haematuria caused by traumatic renal arterio-venous (AV) fistula and pyelocalical injury who was managed by supraselective embolization of the irrigating arterial vessel. CONCLUSION: Supraselective embolization is a first choice method for the treatment of low grade renal vessel injuries. Even patients with the most severe renovascular lesions and major renal destruction (a grade IV injury) can be treated nonsurgically with supraselective embolization, with an excellent chance to preserve the maximal amount of functional renal parenchyma. This method is rapid, effective, tissue preserving, and likely to reduce morbidity and mortality. PMID- 17121385 TI - [Bilobular pancreas: another variant of the divided pancreas?]. AB - BACKGROUND: Pancreas divisum is the most common anomaly of the pancreas. This anomaly has been known as a possible cause of recurrent pancreatitis. CASE REPORT: We performed computerized tomography (CT) of the abdomen in 5 children in whom a divided pancreas was confirmed using endoscopic cholangiopancreatography. In a girl, who had three episodes of severe acute pancreatitis, a CT examination confirmed a completely divided embryonal dorsal and ventral primordium. We named this variant of the divided pancreas the "bilobular pancreas". Contrary to the remaining 4 children in whom the control of the number and severity of attacks, as well as the control of pancreatic pain were achieved by pharmacotherapeutics and an adequate diet, in the reported patient sphincteroplasty of the papilla duodeni minor resulted in a full control of the disease. CONCLUSION: The paper discussed the possibility that the variant of the divided pancreas, with anatomically completely separated ventral and dorsal pancreas and their ductal systems, is the key factor that determines the severity of pancreatic disease and an indication for sphincteroplasty of the papilla duodeni minor as the major therapeutic method. PMID- 17121386 TI - [Prevention of spreading of avian influenza in medical care facilities]. PMID- 17121387 TI - A morphologic and cytochemical study on the great alveolar cell. AB - Lungs from marsupials, bats and rodents were studied by light and electron microscopy. In all three groups, the great alveolar cells exhibit similar morphologic and cytochemical characteristics. Cytoplasmic vacuoles seen in these cells by light microscopy correspond to cytosomes that are demonstrable in them by electron microscopy. Such cytosomes are osmiophilic, periodic acid-Schiff positive and stainable with Sudan black after acetone extraction. After fixation in a mixture of aldehydes, followed by extraction in chloroform-methanol and postfixation in osmium tetroxide, cytosomes lose their osmiophilia. The cytoplasm of the great alveolar cell is notable for a loosely ordered granular endoplasmic reticulum, an extensive Golgi apparatus and numerous multivesicular bodies. Many forms transitional in appearance between multivesicular bodies and cytosomes are present. In these, osmiophilic matter occupies the intervesicular space. It is proposed that these bodies are the precursors of cytosomes. The cytosomes are interpreted to be products of the "lysosomal" system in this cell. Ultimately they are secreted onto the alveolar surface. PMID- 17121388 TI - Evaluation of the glycogenolytic effect of alpha-amylase using radioautography and electron microscopy. AB - The effectiveness of crystalline alpha-amylase and saliva in hydrolyzing newly formed glycogen in liver and muscle was examined. Glycogen synthesis was induced by the administration of H3-glucose to fasting rats or by the incubation of tissue slices in a medium containing H3-glucose. Paraffin sections of Rossman fixed tissues or small pieces of liver fixed in glutaraldehyde and subsequently postosmicated and embedded in Epon were then enzymatically digested. Grain counts were made in radioautographs of treated and untreated materials, and the amount of radioactivity removed by the digestion was used to assess the efficiency of the enzymes in hydrolyzing glycogen. Crystalline alpha-amylase hydrolyzed almost completely newly formed glycogen in liver and muscle. Saliva removed the glycogen that was synthesized in vivo, but it was less effective in hydrolyzing glycogen synthesized in vitro. Electron micrographs of digested liver cells confirmed the radioautographic findings on the effectiveness of the enzyme preparations. PMID- 17121389 TI - Histochemical demonstration of phospholipase B (lysolecithinase) activity in rat tissues. AB - A method has been developed for the histochemical demonstration of phospholipase B (lysolecithinase) of rat tissues. The enzyme attacks lysolecithin with liberation of 1 mole of glycerylphosphorylcholine and 1 mole of fatty acid. The recommended procedure involves use of 6-10 micro frozen sections, fixed in cold calcium-formol and incubated at 37 degrees C in Tris buffered medium at pH 6.6 containing 2.2 X 10(-3) M lysolecithin and 1% cobalt acetate. The fatty acid liberated by enzymatic hydrolysis is trapped as a cobalt precipitate and is then converted to a black-brown precipitate by treatment with dilute ammonium sulfide in cold isotonic saline. Equivalent amounts of fatty acid and glycerylphosphorylcholine are recovered by extraction and analysis of the incubated sections and of the incubation medium, thus proving that lysolecithin hydrolysis occurs under the proposed reaction conditions. Staining is reduced by treating the sections with copper ions, mercury compounds, alcohols, acetone and by heating at 60 degrees C prior to incubation with substrate. Lowering of the pH of the incubation medium has similar effect. These findings are interpreted as evidence of the enzymatic nature of the reaction. Cells exhibiting a positive staining are found in the lamina propria of the intestinal villi and crypts, in the red pulp of the spleen and in the interstitial tissue of lung, liver and thymus. Similar elements are present in bone marrow smears and in leukocyte preparations obtained by peritoneal lavage. The morphologic and staining characteristics of these cells correspond to those of the eosinophilic leukocytes. Physical and chemical agents (x-irradiation, corticosteroids) which sharply decrease the number of eosinophils also reduce the number of cells shown histochemically to hydrolyze lysolecithin. A correspondent diminution of phospholipase B activity of homogenates of the same tissues can be shown in vitro. Differences in tissue distribution and chemical properties distinguish the phospholipase B from less specific esterases and lipases. PMID- 17121390 TI - The number of acetylcholinesterase molecules in the rat megakaryocyte. AB - A megakaryocyte cell series from rat bone marrow has been examined by the isotopic di-isopropyl fluorophosphate (DFP) method for esterases. After complete reaction with 32P-DFP, the numbers of DFP-reacted molecules inindividual cells havebeen determined by beta trackauto-radiography. Previous work has shown the percentage of organophosphate-sensitive sites in these cells which can be taken as active centers of acetylcholinesterase (AChase). Combining these data, the absolute numbers of organophosphate-sensitive esterase molecules and AChase molecules per cell were determined. Histograms show a narrow spread of values within each of four size classes from megakaryoblast to fully mature megakaryocyte, but, with means increasing 4-fold through this series, approximately in proportion to cell volume. A rat megakaryoblast has 2 X 10(6) AChase molecules, and a megakaryocyte (of 48-micro diameter) has 7.6 X 10(6) molecules. The apparent turnover number of the enzyme for intracellular reaction with substrate is calculated and compared with turnover numbers available for other AChases. PMID- 17121391 TI - A semiautomatic instrument for the radioautographic coating technique. AB - By means of a mechanical coating instrument a fast, simple method to coat specimens with liquid nuclear track emulsion has been devised for quantitative light and electron microscopic radioautography. In both cases, the section is mounted on a glass slide. After the vertically held slide has been immersed in the melted emulsion, the instrument withdraws it at a slow, constant speed. As a result, the specimen is coated with a thin, uniform emulsion layer composed of homogeneously distributed silver bromide crystals. The thickness of the emulsion coat may be standardized by selection of an optimal combination of emulsion dilution, temperature and withdrawal speed. PMID- 17121392 TI - Enzyme-labeled antibodies: preparation and application for the localization of antigens. PMID- 17121393 TI - Ultrastructural localization of aryl sulfatase activity in plant meristematic cells. PMID- 17121394 TI - Demonstration of 5'-nucleotidase activity in decalcified bones and teeth. PMID- 17121395 TI - Shape dependence in the formation of condensed phases exhibited by disubstituted sucrose esters. AB - We report on the self-organizing properties of sucrose esters that are di-(1',6', 1',6, and 6,6')-substituted with aliphatic chains of identical or different chain lengths and levels of saturation. For the materials possessing two saturated aliphatic chains, the compounds exhibited thermotropic lamellar smectic A phases. A remarkable new phase transition was observed for the di-octadecanoyl homologue in which one smectic A phase transformed into another with a continuous change in layer spacing, but with a discontinuous change in the correlation length. The incorporation of long cis-unsaturated chains led to increased cross-sectional areas of the chains relative to the sucrose head groups and, hence, columnar phases were observed. PMID- 17121396 TI - Unexpected conformational properties of 1-trifluoromethyl-1-silacyclohexane, C5H10SiHCF3: gas electron diffraction, low-temperature NMR spectropic studies, and quantum chemical calculations. AB - The molecular structure of axial and equatorial conformers of 1-trifluoromethyl-1 silacyclohexane, (C5H10SiHCF3), as well as the thermodynamic equilibrium between these species was investigated by means of gas electron diffraction (GED), dynamic nuclear magnetic resonance (DNMR) spectroscopy, and quantum chemical calculations (B3LYP, MP2, and CBS-QB3). According to GED, the compound exists as a mixture of two Cs symmetry conformers possessing the chair conformation of the six-membered ring and differing in the axial or equatorial position of the CF3 group (axial=58(12) mol%/equatorial=42(12) mol%) at T=293 K. This result is in a good agreement with the theoretical prediction. This is, however, in sharp contrast to the conformational properties of the cyclohexane analogue. The main structural feature for both conformers is the unusually long exocyclic bond length Si--C 1.934(10) A. A low-temperature 19F NMR experiment results in an axial/equatorial ratio of 17(2) mol%:83(2) mol% at 113 K and a DeltaG (not equal) of 5.5(2) kcal mol-1. CBS-QB3 calculations in the gas-phase and solvation effect calculations using the PCM(B3LYP/6-311G*) and IPCM(B3LYP/6-311G*) models were applied to estimate the axial/equatorial ratio in the 100-300 K temperature range, which showed excellent agreement with the experimental results. The minimum energy pathways for the chair-to-chair inversion of trifluoromethylsilacyclohexane and methylsilacyclohexane were also calculated using the STQN(Path) method. PMID- 17121397 TI - Metal-ion-coordinating properties of the dinucleotide 2'-deoxyguanylyl(5'-->3') 2'-deoxy-5'-guanylate (d(pGpG)3-): isomeric equilibria including macrochelated complexes relevant for nucleic acids. AB - The interaction between divalent metal ions and nucleic acids is well known, yet knowledge about the strength of binding of labile metal ions at the various sites is very scarce. We have therefore studied the stabilities of complexes formed between the nucleic acid model d(pGpG) and the essential metal ions Mg2+ and Zn2+ as well as with the generally toxic ions Cd2+ and Pb2+ by potentiometric pH titrations; all four ions are of relevance in ribozyme chemistry. A comparison of the present results with earlier data obtained for M(pUpU)- complexes allows the conclusion that phosphate-bound Mg2+ and Cd2+ form macrochelates by interaction with N7, whereas the also phosphate-coordinated Pb2+ forms a 10-membered chelate with the neighboring phosphate diester bridge. Zn2+ forms both types of chelates with formation degrees of about 91% and 2.4% for Zn[d(pGpG)]cl/N7 and Zn[d(pGpG)] cl/PO, respectively; the open form with Zn2+ bound only to the terminal phosphate group, Zn[d(pGpG)]-op, amounts to about 6.8 %. The various intramolecular equilibria have also been quantified for the other metal ions. Zn2+, Cu2+, and Cd2+ also form macrochelates in the monoprotonated M[H;d(pGpG)] species (the proton being at the terminal phosphate group), that is, the metal ion at N7 interacts to some extent with the P(O)2(OH)- group. Thus, this study demonstrates that the coordinating properties of the various metal ions toward a pGpG unit in a nucleic acid differ: Mg2+, Zn2+, and Cd2+ have a significant tendency to bridge the distance between N7 and the phosphate group of a (d)GMP unit, although to various extents, whereas Pb2+ (and possibly Ca2+) prefer a pure phosphate coordination. PMID- 17121398 TI - [40]nonaphyrin(1.1.1.1.1.1.1.1.1) and its heterometallic complexes with palladium carbon bonds. AB - meso-Pentafluorophenyl- substituted [40]nonaphyrin(1.1.1.1.1.1.1.1.1) 3 has been prepared by using a stepwise ring-size-selective synthesis, and has been reduced with NaBH(4) to [42]nonaphyrin(1.1.1.1.1.1.1.1.1) 5. Structurally, 3 is characterized by a figure-of-eight shape, consisting of a porphyrin-like tetrapyrrolic segment and a hexaphyrin-like hexapyrrolic segment, whereas 5 has been found to adopt a distorted nonplanar butterfly-like shape. In the mono-metal complexes 6 and 7, a Zn(II) or Cu(II) ion is bound by the porphyrin-like tetrapyrrolic segment, maintaining the overall structure of 3. Similarly to 3, complexes 6 and 7 are interconvertible with the corresponding complexes 9 and 10 through two-electron reduction with NaBH(4) and oxidation with DDQ. The metal free hexaphyrin-like segments of 6 and 7 have been shown to serve as a suitable platform for the complexation of two palladium ions, providing hetero-trinuclear metal complexes 11 (Zn(II)-Pd(II)-Pd(II)) and 13 (Cu(II)-Pd(II)-Pd(II)) in high yields, in which the Zn or Cu ion resides at the same porphyrin-like segment, and one Pd ion is bound in an NNCC fashion through double C--H bond activation while the other is bound in an NNC fashion with single C--H bond activation. Multi metal complexes 11, 12, and 13 exhibit small electrochemical HOMO-LUMO gaps (<0.6 eV), despite their nonplanar conformations. PMID- 17121399 TI - Hydrogen storage in the giant-pore metal-organic frameworks MIL-100 and MIL-101. PMID- 17121400 TI - Induction of structure and function in a designed peptide upon adsorption on a silica nanoparticle. PMID- 17121401 TI - Dancing on a fullerene surface: isomerization of Y(3)N@(N-ethylpyrrolidino-C(80)) from the 6,6 to the 5,6 regioisomer. PMID- 17121402 TI - Microtiter plate-based screening for the optimization of DNA-protein conjugate synthesis by means of expressed protein ligation. AB - We report a rapid microtiter plate screening assay for the optimization of the synthesis of covalent DNA-protein conjugates by means of expressed protein ligation (EPL). The EPL method allows for the site-specific coupling of cysteine modified DNA oligomers with recombinant intein-fusion proteins, the latter containing a C-terminal thioester that enables a mild and highly specific reaction with N-terminal cysteine compounds. To screen for optimal reaction conditions, we developed a microtiter plate-based assay that utilizes DNA directed immobilization of the products formed in the ligation reaction of cysteine-modified DNA oligonucleotides with the model protein thioester of the maltose-binding protein (MBP), recombinantly expressed as an intein-fusion protein in E. coli. The screening assay allowed the rapid quantitative monitoring of various reaction parameters, such as the ratio of the reactants, reaction times, pH and ion strength of the buffer, the influence of various thiol additives and the nature of the chemical linker within the cysteine-bearing DNA oligonucleotide. As the consequence of the assay-based optimization, the ligation of MBP with the oligonucleotide was improved to near quantitative yields. PMID- 17121403 TI - A molecular probe for the detection of homopurine sequences. PMID- 17121404 TI - Synthesis of [1,2-13C2, 15N]-L-homoserine and its incorporation by the PKS-NRPS system of Fusarium moniliforme into the mycotoxin fusarin C. PMID- 17121405 TI - Functional immobilization of the small GTPase Rab6A on DNA-Gold nanoparticles by using a site-specifically attached poly(ethylene glycol) linker and thiol place exchange reaction. PMID- 17121406 TI - Developments in the characterisation of the catalytic triad of alpha chymotrypsin: Effect of the protonation state of Asp102 on the 1H NMR signals of His57. PMID- 17121407 TI - Hyperfine coupling tensors of the benzosemiquinone radical anion from Car Parrinello molecular dynamics. AB - Based on Car-Parrinello ab initio molecular dynamics simulations of the benzosemiquinone radical anion in both aqueous solution and the gas phase, density functional calculations provide the currently most refined EPR hyperfine coupling (HFC) tensors of semiquinone nuclei and solvent protons. For snapshots taken at regular intervals from the molecular dynamics trajectories, cluster models with different criteria for inclusion of water molecules and an additional continuum solvent model are used to analyse the HFCs. These models provide a detailed picture of the effects of dynamics and of different intermolecular interactions on the spin-density distribution and HFC tensors. Comparison with static calculations allows an assessment of the importance of dynamical effects, and of error compensation in static DFT calculations. Solvent proton HFCs depend characteristically on the position relative to the semiquinone radical anion. A point-dipolar model works well for in-plane hydrogen-bonded protons but deviates from the quantum chemical values for out-of-plane hydrogen bonding. PMID- 17121408 TI - On the origin of red and blue shifts of X-H and C-H stretching vibrations in formic acid (formate ion) and proton donor complexes. AB - Complexes between formic acid or formate anion and various proton donors (HF, H(2)O, NH(3), and CH(4)) are studied by the MP2 and B3LYP methods with the 6 311++G(3df,3pd) basis set. Formation of a complex is characterized by electron density transfer from electron donor to ligands. This transfer is much larger with the formate anion, for which it exceeds 0.1 e. Electron-density transfer from electron lone pairs of the electron donor is directed into sigma* antibonding orbitals of X--H bonds of the electron acceptor and leads to elongation of the bond and a red shift of the X--H stretching frequency (standard H-bonding). However, pronounced electron-density transfer from electron lone pairs of the electron donor also leads to reorganization of the electron density in the electron donor, which results in changes in geometry and vibrational frequency. These changes are largest for the C--H bonds of formic acid and formate anion, which do not participate in H-bonding. The resulting blue shift of this stretching frequency is substantial and amounts to almost 35 and 170 cm(-1), respectively. PMID- 17121409 TI - Microsolvation of an ionic dopant in small (4)He clusters: OH(+)((3)sigma)((4)He)(N) via genetic algorithm optimizations. AB - The optimized spatial structures of the small clusters (with N up to 33) formed by an increasing number of (4)He atoms, which act as a microsolvent surrounding the OH(+) ionic molecular dopant, are obtained using a sum-of-potentials scheme corrected by three-body (3B) effects. The most stable structures are generated using the type of genetic algorithm described herein, and the sequential formation of regular shell structures is analyzed in detail. Possible quantum corrections for both the solvent distributions and the stable energetics are analyzed and discussed. PMID- 17121410 TI - pH sensitivity of Si--C linked organic monolayers on crystalline silicon surfaces. AB - The electrochemical behavior of Si--C linked organic monolayers is studied in electrolyte-insulator-Si devices, under conditions normally encountered in potentiometric biosensors, to gain fundamental knowledge on the behavior of such Si electrodes under practical conditions. This is done via titration experiments, Mott-Schottky data analysis, and data fitting using a site-binding model. The results are compared with those of native SiO(2) layers and native SiO(2) layers modified with hexamethyldisilazane. All samples display pH sensitivity. The number of Si--OH groups on the alkylated samples is calculated to be less than 0.7 % of that of a pure SiO(2) insulator, which still causes a pH sensitivity of approximately 25 mV per pH unit in the pH range: 4-7. The alkylated samples hardly suffer from response changes during up- and down-going titrations, which indicates that very little oxide is additionally formed during the measurements. The pK(a) values of all samples with monolayers (4.0-4.4) are lower than that of native SiO(2) (6.0). The long-term drift (of approximately 1 mV h(-1)) is moderate. The results indicate that biosensors composed of alkylated Si substrates are feasible if a cross-sensitivity towards pH in the sensor signal is taken into account. PMID- 17121411 TI - Eigen or Zundel ion: news from calculated and experimental photoelectron spectroscopy. PMID- 17121412 TI - Local structure and thermodynamics of a core-softened potential fluid: theory and simulation. AB - Phase behavior and structural properties of homogeneous and inhomogeneous core softened (CS) fluid consisting of particles interacting via the potential, which combines the hard-core repulsion and double attractive well interaction, are investigated. The vapour-liquid coexistence curves and critical points for various interaction ranges of the potential are determined by discrete molecular dynamics simulations to provide guidance for the choice of the bulk density and potential parameters for the study of homogeneous and inhomogeneous structures. Spatial correlations in the homogeneous CS system are studied by the Ornstein Zernike integral equation in combination with the modified hypernetted chain (MHNC) approximation. The local structure of CS fluid subjected to diverse external fields maintaining the equilibrium with the bulk CS fluid are studied on the basis of a recently proposed third order+second order perturbation density functional approximation (DFA). The accuracy of DFA predictions is tested against the results of a grand canonical ensemble Monte Carlo simulation. Reasonable agreement between the results of both methods proves that the DFA theory applied in this work is a convenient theoretical tool for the investigation of the CS fluid, which is practically applicable for modeling numerous real systems. PMID- 17121413 TI - Quantification of intracellular homocysteine by stable isotope dilution liquid chromatography/tandem mass spectrometry. AB - A precise and accurate stable isotope dilution liquid chromatography/tandem mass spectrometry method for the analysis of intracellular homocysteine has been developed. An internal standard, [(2)H(8)]-homocystine, was added to cell pellets from EA.hy 926 cells grown in culture under low and high folate concentrations. D,L-dithiothreitol was used to reduce cellular homocystine to homocysteine. Cellular proteins were precipitated by the addition of formic acid in acetonitrile. After centrifugation, a portion of the supernatant was analyzed by liquid chromatography/tandem mass spectrometry. Using a Supelcosil cyano column with an Applied Biosystems API 4000 triple quadrupole mass spectrometer, the SRM transitions for homocysteine (m/z 136 to m/z 90) and [(2)H(4)]-homocysteine (m/z 140 to m/z 94) were monitored. The method was validated by conducting five replicate analyses on three different days at four different concentrations (concentrations at the lower limit of quantitation and expected lower quartile, mid-range and upper quartile). The limit of detection was 2 ng/10(6) EA.hy 926 cells. Using this method, the intracellular homocysteine concentration in EA.hy 926 cells ranged from 10 to 36 ng/10(6) cells. PMID- 17121414 TI - Correlation between prenatal ultrasound and autopsy findings: A study of second trimester abortions. AB - OBJECTIVE: To evaluate discrepancies between sonographic and autopsy findings following termination of pregnancy (TOP) in the second trimester. METHODS: This retrospective report is based on 288 consecutive second-trimester abortions carried out because of fetal malformations diagnosed by ultrasound examination at a tertiary referral center. The correlation between the results from the ultrasound and autopsy examinations was evaluated. RESULTS: Autopsy was performed in 274 cases. In 160 of the 274 pregnancies (58.4%) there was full agreement between the two examination methods. Findings detected by autopsy (in addition to those leading to termination) were not observed by ultrasonography in 86 (31.4%) of the pregnancies; of the 64 malformations that occurred, 30 (46.9%) were judged as 'detectable'. In 27 (9.9%) pregnancies, observations made by ultrasound (in addition to those leading to termination) were not confirmed at autopsy. In one pregnancy, postmortem radiology examination-but not autopsy-confirmed the ultrasound observations. No pregnancies were terminated because of false positive ultrasound observations. The correlation between ultrasound and autopsy findings was evaluated by three investigators; the inter-rater agreement was high (kappa = 0.85). CONCLUSION: Discrepancies between ultrasound and autopsy findings were observed in about 40% of the pregnancies. These discrepancies confirm the need for autopsy following TOP. PMID- 17121415 TI - Mind the gap! What we don't know about right aortic arches and aberrant branches. PMID- 17121416 TI - Towards the minimally invasive autopsy? PMID- 17121418 TI - Deficiency of tenascin-C attenuates liver fibrosis in immune-mediated chronic hepatitis in mice. AB - Tenascin-C (TNC), an extracellular matrix glycoprotein, is upregulated in chronic liver disease. Here, we investigated the contribution of TNC to liver fibrogenesis by comparing immune-mediated hepatitis in wild-type (WT) and TNC null (TNKO) mice. Eight-week-old BALB/c mice received weekly intravenous injections of concanavalin A to induce hepatitis, and were sacrificed one week after the 3rd, 6th, 9th, and 12th injections. In WT livers, immunohistochemical staining revealed a gradual increase in TNC deposition. TNC mRNA levels also increased sequentially and peaked after the 9th injection. Collagen deposition, stained with picrosirius red, was significantly less intense in TNKO mice than in WT mice, and procollagen I and III transcripts were significantly upregulated in WT mice compared with TNKO mice. Inflammatory infiltrates were most prominent after the 3rd-6th injections in both groups and were less intense in TNKO mice than in WT mice. Interferon-gamma, tumour necrosis factor-alpha, and interleukin 4 mRNA levels were significantly higher in WT mice than in TNKO mice, while activated hepatic stellate cells (HSCs) and myofibroblasts, a cellular source of TNC and procollagens, were more common in WT livers. Transforming growth factor (TGF)-beta1 mRNA expression was significantly upregulated in WT mice, but not in TNKO mice. In conclusion, TNC can promote liver fibrogenesis through enhancement of inflammatory response with cytokine upregulation, HSC recruitment, and TGF beta expression during progression of hepatitis to fibrosis. PMID- 17121419 TI - Identification of the nitration site of insulin by peroxynitrite. AB - Our previous investigation indicated that insulin can be nitrated by peroxynitrite in vitro. In this study, the preferential nitration site of the four tyrosine residues in insulin molecule was confirmed. Mononitrated and dinitrated insulins were purified by RP-HPLC. Following reduction of insulin disulfide bridges, Native-PAGE indicated that A-chain was preferentially nitrated. Combination of enzymatic digestion of mononitrated insulin with endoproteinase Glu-C, mass spectrometry confirmed that Tyr-A14 was the preferential nitration site when insulin was treated with peroxynitrite. Tyr-A19, maybe, was the next preferential nitration site. According to the crystal structure, Tyr-B26 between the two tyrosine residues in insulin B-chain was likely easier to be nitrated by peroxynitrite. PMID- 17121420 TI - Limiting racemization and aspartimide formation in microwave-enhanced Fmoc solid phase peptide synthesis. AB - Microwave energy represents an efficient manner to accelerate both the deprotection and coupling reactions in 9-fluorenylmethyloxycarbonyl (Fmoc) solid phase peptide synthesis (SPPS). Typical SPPS side reactions including racemization and aspartimide formation can occur with microwave energy but can easily be controlled by routine use of optimized methods. Cysteine, histidine, and aspartic acid were susceptible to racemization during microwave SPPS of a model 20mer peptide containing all 20 natural amino acids. Lowering the microwave coupling temperature from 80 degrees C to 50 degrees C limited racemization of histidine and cysteine. Additionally, coupling of both histidine and cysteine can be performed conventionally while the rest of the peptide is synthesized using microwave without any deleterious effect, as racemization during the coupling reaction was limited to the activated ester state of the amino acids up to 80 degrees C. Use of the hindered amine, collidine, in the coupling reaction also minimized formation of D-cysteine. Aspartimide formation and subsequent racemization of aspartic acid was reduced by the addition of HOBt to the deprotection solution and/or use of piperazine in place of piperidine. PMID- 17121421 TI - Solid-phase synthesis and purification of a set of uniformly 13C, 15N labelled de novo designed membrane fusogenic peptides. AB - The transmembrane segments of soluble N-ethylmaleimide-sensitive factor (SNARE) proteins or viral envelope proteins drive membrane fusion, which suggests that simple synthetic biology constructs for fusion exist and can be evaluated. We describe the high-yield synthesis of a set of de novo designed fusogenic peptides for use in functional investigations, which are highly enriched in 13C and 15N using three equivalents of labelled amino acids and optimized reaction conditions minimizing aggregation. The biomimetic peptides have a high purity >90% and show reproducible and fusogenic activity that correlates well with the intended functional design characteristics, from strongly fusogenic to almost non fusogenic. PMID- 17121422 TI - Sonographic detection of fetal cerebellar cavernous hemangioma with in-utero hemorrhage leading to cerebellar hemihypoplasia. AB - We report the prenatal diagnosis of cerebellar cavernous angioma. Ultrasound examination at 21 weeks' gestation showed a hyperechogenic lesion measuring 12 mm in diameter, occupying most of the right cerebellar hemisphere. At 24 weeks' gestation, significant hypoplasia of the right hemisphere was diagnosed, and the pregnancy was terminated. Pathological evaluation of the lesion revealed extensive hemorrhaging and a rich network of dilated vessels and small capillaries. The right cerebellar lobe was hypoplastic and covered with blood clots. Cerebral cavernous malformation should be considered in the differential diagnosis of hyperechogenic lesions in the fetal cerebellum. PMID- 17121423 TI - Three-dimensional sonographic determination of normal fetal mandibular and maxillary size during the second half of pregnancy. AB - OBJECTIVE: To explore the various ways of obtaining fetal maxillary and mandibular size with three-dimensional (3D) ultrasound, with a view to developing a tool for identifying minor anomalies in the lower facial region. METHODS: Serial 3D sonographic measurements of the fetal jaws were made in 126 normal singleton pregnancies at 18-34 weeks of gestation for determination of degree of maxillary and mandibular protrusion, maxillary and mandibular corpus lengths, mid and lower facial depths and maxillary and mandibular curvature. In a sub study the reproducibility of the measurements was evaluated. RESULTS: The coefficient of variation in the reproducibility study varied between 7.1 and 10.5%. For all parameters except maxillary and mandibular protrusion, there was a significant gestational age-related increase. Ratios of maxillary/mandibular protrusion, maxillary/mandibular corpus lengths, mid-/lower facial depths and maxillary/mandibular curvature all showed a significant gestational age-related decrease, with the most distinct decrease in the mid-/lower facial depth ratio. CONCLUSIONS: 3D sonographic measurement of the fetal maxilla and mandible demonstrated an acceptable intraobserver variability for all measurements. The mid-/lower facial depth ratio appears to be most valuable in determining abnormal mandibular development. PMID- 17121424 TI - The moving face of the fetus-the changing face of medicine. PMID- 17121425 TI - Re: Second-trimester intra-abdominal bowel dilation in fetuses with gastroschisis predicts neonatal bowel atresia. PMID- 17121426 TI - Subtelomeric rearrangements and 22q11.2 deletion syndrome in anomalous growth restricted fetuses with normal or balanced G-banded karyotype. AB - OBJECTIVE: To determine the frequencies of cryptic subtelomeric rearrangements and 22q11.2 deletion in anomalous growth-restricted fetuses with normal or balanced G-banded karyotypes. METHODS: This was a study of 27 consecutive fetuses at a median gestational age of 26 (range, 19-33) weeks, that had intrauterine growth restriction (IUGR) as well as at least one major structural anomaly, and a normal or balanced G-banded karyotype. The median maternal age was 29 (range, 17 39) years. Fluorescence z in-situ hybridization (FISH) diagnosis of the cultured amniocytes with the probe TUPLE 1, and then the Chromoprobe Multiprobe-T system were used, respectively, to screen for the frequency of 22q11.2 deletion syndrome and subtelomeric rearrangements involving the 41 unique chromosome ends (i.e. excluding the five short arms of acrocentric autosomes (no. 13, 14, 15, 21 and 22)). Those that had suspected deleted subtelomeres were reanalyzed with a specific subtelomeric probe, TelVysion. RESULTS: Of the 27 fetuses, three (11%) were affected with 22q11.2 deletion syndrome and two (7.4%) had subtelomeric deletions (one monosomy 21q22.3, one monosomy 1p36.3). Of the 11 fetuses with congenital heart defects, three (27.3%) had 22q11.2 deletion syndrome and one (9.1%) had monosomy 1p36.3. In the remaining 16 fetuses without congenital heart defects, none had 22q11.2 deletion syndrome. However, one (6.3%) had cryptic rearrangement involving subtelomeres. CONCLUSION: Prenatal subtelomeric FISH screening is technically feasible using cultured amniocytes. We propose that 22q11.2 deletion syndrome and cryptic subtelomere rearrangements may be important etiologies of fetuses with IUGR and at least one structural anomaly, along with a normal karyotype or one that is balanced by traditional G-banding. Fetuses with congenital heart defects and IUGR should undergo FISH to exclude 22q11.2 deletion syndrome. In fetuses with IUGR and at least one major structural anomaly but without congenital heart defects, screening of subtelomeric rearrangements may contribute to further elucidation of the underlying etiology. PMID- 17121427 TI - Test characteristics of placental location screening by transabdominal ultrasound at 18-20 weeks. AB - OBJECTIVES: To determine the test characteristics of a second-trimester transabdominal fetal anomaly scan in screening for placenta previa. PATIENTS AND METHODS: This was a case-control study carried out at a tertiary obstetric hospital involving 54 women (0.5% of the total number booked for antenatal care and delivery at the center) with clinically significant placenta previa at the time of delivery, and 168 randomly selected controls who attended for antenatal care and delivery in 2000-2003. Sensitivity, specificity and false positive rate of the placenta-os measurement in the second-trimester transabdominal fetal anomaly scan for identification of clinically significant placenta previa were assessed. RESULTS: Women with placenta previa at delivery had significantly different second-trimester placenta-os measurements from those of controls (P < 0.0001). For women with placenta previa, the mean (standard deviation) placenta os measurement was -0.96 cm (1.95) vs. 4.15 cm (1.77) for controls (P < 0.0001). Using the anomaly scan as a screening test for placenta previa with a placenta-os measurement cut-off point of < or = 2 cm, all cases of placenta previa should be identified, but 11.1% (95% CI, 7.1-14.8) of women without placenta previa would have false positive results. At cut-off points of 3 cm and 1.5 cm the false positive rates were 25.7% (95% CI, 18.8-30.2%) and 6.6% (95% CI, 3.8-9.6%), respectively, but at 1.5 cm 3.8% of cases would be missed. CONCLUSIONS: The second-trimester transabdominal fetal anomaly scan is a useful screening test for placenta previa. However, because of false positives among the much larger population of women without placenta previa, we estimate that with a cut-off point of < or =2 cm, 23 women would have to be followed up to confirm one true placenta previa. PMID- 17121428 TI - The CADEUS study: methods and logistics. AB - PURPOSE: At the request of the French Health authorities, a study called CADEUS (COX-2 inhibitors and NSAIDs: description of users) aimed to describe the users of cyclo-oxygenase (COX)-2 inhibitors and traditional non-selective non-steroidal anti-inflammatory drugs (tNSAIDs). We report here the methodology, logistics and study design performances. METHODS: CADEUS is a cohort study designed to include 40,000 patients randomly sampled monthly in the French National Healthcare Insurance database, who received at least one dispensation of celecoxib, rofecoxib or tNSAIDs (1:1:2), from September 2003 to August 2004. Patients and prescribers were asked to fill a questionnaire on indication, medical history, risk factors and hospitalizations since drug acquisition. There was no reminder. For each respondent, healthcare resources used for the 6 months before and after inclusion were extracted from the database. Response rate, response delay, responders and non-responders characteristics were assessed. RESULTS: Of the 222,879 patients and their prescribers contacted, 20.8% patients and 32.6% prescribers responded. Median response delay was 16 days for patients and 17 days for physicians. Factors associated with patient response were age, cohort, type of prescriber and period of inclusion. CONCLUSION: This is the first study of this design in France, combining data from a claims database and direct patient and prescriber questionnaires. PMID- 17121429 TI - Comparison of the global gene expression profiles produced by methylparaben, n butylparaben and 17beta-oestradiol in MCF7 human breast cancer cells. AB - Since the alkyl esters of p-hydroxybenzoic acid (parabens) can be measured intact in the human breast and possess oestrogenic properties, it has been suggested that they could contribute to an aberrant burden of oestrogen signalling in the human breast and so play a role in the rising incidence of breast cancer. However, although parabens have been shown to regulate a few single genes (reporter genes, pS2, progesterone receptor) in a manner similar to that of 17beta-oestradiol, the question remains as to the full extent of the similarity in the overall gene profile induced in response to parabens compared with 17beta oestradiol. The GE-Amersham CodeLink 20 K human expression microarray system was used to profile the expression of 19881 genes in MCF7 human breast cancer cells following a 7-day exposure to 5 x 10(-4) M methylparaben, 10(-5) M n-butylparaben and 10(-8) M 17beta-oestradiol. At these concentrations, the parabens gave growth responses in MCF7 cells of similar magnitude to 17beta-oestradiol. The study identified genes which are upregulated or downregulated to a similar extent by methylparaben, n-butylparaben and 17beta-oestradiol. However, the majority of genes were not regulated in the same way by all three treatments. Some genes responded differently to parabens from 17beta-oestradiol, and furthermore, differences in expression of some genes could be detected even between the two individual parabens. Therefore, although parabens possess oestrogenic properties, their mimicry in terms of global gene expression patterns is not perfect and differences in gene expression profiles could result in consequences to the cells that are not identical to those following exposure to 17beta-oestradiol. PMID- 17121430 TI - Dissection of signaling pathways in fourteen breast cancer cell lines using reverse-phase protein lysate microarray. AB - Signal transduction pathways play a crucial role in breast cancer development, progression, and response to different therapies. A major problem in breast cancer therapy is the heterogeneity among different tumor types and cell lines commonly used in preclinical studies. To characterize the signaling pathways of some of the commonly used breast cancer cell lines and dissect the relationship among a number of pathways and some key genetic and molecular events in breast cancer development, such as p53 mutation, ErbB2 expression, and estrogen receptor (ER)/progesterone receptor (PR) status, we performed pathway profiling of 14 breast cancer cell lines by measuring the expression and phosphorylation status of 40 different cell signaling proteins with 53 specific antibodies using a protein lysate array. Cluster analysis of the expression data showed that there was close clustering of phosphatidylinositol 3-kinase, Akt, mammalian target of rapamycin (mTOR), Src, and platelet-derived growth factor receptor beta (PDGFRbeta) in all of the cell lines. The most differentially expressed proteins between ER- and PR-positive and ER- and PR-negative breast cells were mTOR, Akt (pThr308), PDGFRbeta, PDGFRbeta (pTyr751), panSrc, Akt (pSer473), insulin-like growth factor-binding protein 5 (IGFBP5), Src (pTyr418), mTOR (pSer2448), and IGFBP2. Many apoptotic proteins, such as apoptosis-inducing factor, IGFBP3, bad, bax, and cleaved caspase 9, were overexpressed in mutant p53-carrying breast cancer cells. Hexokinase isoenzyme 1, ND2, and c-kit were the most differentially expressed proteins in high and low ErbB2-expressing breast cancer cells. This study demonstrated that ER/PR status, ErbB2 expression, and p53 status are major molecules that impact downstream signaling pathways. PMID- 17121431 TI - Pharmacogenomics of the polyamine analog 3,8,13,18-tetraaza-10,11-[(E)-1,2 cyclopropyl]eicosane tetrahydrochloride, CGC-11093, in the colon adenocarcinoma cell line HCT1161. AB - Polyamine analogs are known to inhibit tumorigenesis at least in part by mimicking some of the regulatory roles of natural polyamines. To begin the identification of those signaling pathways that are involved in differential cellular responses to the synthetic conformationally restricted polyamine analog CGC-11093, we conducted gene expression profiling, proteomic, and genome-wide DNA methylation and histone acetylation analyses of the HCT116 colon adenocarcinoma cell line after treatment with this analog. Gene expression analysis was performed using Affymetrix GeneChip human genome U133 Plus 2.0 arrays. Changes in protein expression were evaluated using 2D polyacrylamide gels followed by LCMS/MS. DNA methylation was measured using 6,800 element CpG island microarrays. Treatment of cells with CGC-11093 at concentrations ranging from 0.1 to 10 microM caused inhibition of cell growth and metabolic activity, but only minimally affected cell viability. Gene expression analysis showed concentration-dependent effects of CGC-11093 on the DNA/RNA binding transcription factor, cell cycle, signaling, transport, cytoskeletal/structural, and serine protease genes. Functional gene analysis revealed distinct expression patterns related to inhibition of cell cycle control, TGF beta signaling, proteasome and RNA polymerase pathways, upregulation of the aminoacyl-tRNA synthesis pathway, and perturbations in the MAPK and Wnt signaling pathways. Microarray results were validated for selected genes with real time RT PCR. Proteomics analysis showed correlative changes in the expression of proteins involved in the regulation of proteasome function (proteasome subunit Y) and tRNA synthesis. CGC-11093 treatment did not produce any detectable changes in DNA methylation or histone acetylation in cells. This study validates specific target pathways for a specific conformationally restricted polyamine analog and suggests the utility of combined gene and DNA methylation microarrays along with proteomic analyses as a useful approach to the evaluation of the mechanisms of action of anticancer drugs. PMID- 17121432 TI - Wnt-1 is dominant over neu in specifying mammary tumor expression profiles. AB - Wnt-1 and Neu collaborate to induce mammary tumors in bitransgenic mice carrying both MMTV-Wnt-1 and MMTV-Neu. In this report, gene expression profiles were determined for tumors from these bitransgenic mice, and compared with expression profiles of tumors from mice singly transgenic for MMTV-Wnt-1 or MMTV-Neu. While very different from tumors arising in MMTV-Neu transgenic mice, tumors from these bitransgenic mice were found not to have identifiable differences from tumors from MMTV-Wnt-1 transgenic mice, using clustering and multidimensional scaling analyses (unsupervised and supervised), One-way Analysis of Variance (ANOVA), and two sample t test (the later two of which were combined with false discovery rate computation). These observations suggest that Wnt-1 is dominant over Neu in specifying mammary tumor expression profiles. PMID- 17121433 TI - Clusterin expression and univariate analysis of overall survival in human breast cancer. AB - The aim of this research is to investigate the significance of clusterin (CLU) expression as a risk factor for breast cancer through tissue microarray technology and univariate analysis of overall survival. Formalin-fixed, paraffin embedded tissues from 158 cases of breast cancer and 31 cases of normal adjacent tissues assembled into a tissue microarray. Cytoplasmic CLU expression in tumor tissues was measured by immunochemistry. Survival analysis was used to investigate the relationship between CLU expression and prognosis, tumor volume, pathological classification, and recurrence. Survival time of patients with CLU expression, lymph node metastasis, and limited post-surgery chemotherapy (<6 cycles of treatment) was significantly shorter than that of patients with no detectable CLU expression (P=0.000), without lymph node metastasis (P=0.000) and more comprehensive post-surgery chemotherapy (>/=6 cycles of treatment) (P=0.035). CLU expression in tumor cells was higher than in normal adjacent breast epithelial cells (P=0.03). The CLU expression staining coefficient of cancer tissues with lymph node metastasis was higher than those without lymph node metastasis (P=0.000). Cytoplasmic CLU expression was found to be a prognostic factor for human breast cancer. PMID- 17121434 TI - Confidence intervals for the true classification error conditioned on the estimated error. AB - Bias and variance for small-sample error estimation are typically posed in terms of statistics for the distributions of the true and estimated errors. On the other hand, a salient practical issue asks, given an error estimate, what can be said about the true error? This question relates to the joint distribution of the true and estimated errors, specifically, the conditional expectation of the true error given the error estimate. A critical issue is that of confidence bounds for the true error given the estimate. We consider the joint distribution of the true error and the estimated error, assuming a random feature-label distribution. From it, we derive the marginal distributions, the conditional expectation of the estimated error given the true error, the conditional expectation of the true error given the estimated error, the conditional variance of the true error given the estimated error, and the 95% upper confidence bound for the true error given the estimated error. Numerous classification and estimation rules are considered across a number of models. Massive simulation is used for continuous models and analytic results are derived for discrete classification. We also consider a breast-cancer study to illustrate how the theory might be applied in practice. Although specific results depend on the classification rule, error-estimation rule, and model, some general trends are seen: (I) if the true error is small (large), then the conditional estimated error is generally high (low)-biased; (II) the conditional expected true error tends to be larger (smaller) than the estimated error for small (large) estimated errors; and (III) the confidence bounds tend to be well above the estimated error for low error estimates, becoming much less so for large estimates. PMID- 17121435 TI - The Role for intensity modulated radiation therapy (IMRT) in pediatric population. AB - The purpose of this study was to evaluate our initial experience on the use of IMRT in children with tumors in eloquent or critical locations. Twenty-two children with a median age of 12 years (range 1-17) were treated using IMRT for tumors which were within 2 cm of a critical structure. The treatment locations were spine [2], head and neck [5], abdominopelvic [8], and intracranial [7]. Eighty-two percent (82%) of patients were treated with curative intent despite most patients having advanced or metastatic disease and two patients having previously received standard external beam radiation. IMRT was delivered with a 6MV linear accelerator using dynamic multileaf collimators with a median of six fields. The median follow-up was five months [1-21]. The median administered dose was 45 Gy. The median planning treatment volume (PTV) was 105.4 cc. For the intracranial lesions, the mean doses to the pituitary, brainstem, cochlea, optic nerve, and lens were 31%, 42%, 17%, 27%, and 6% of the total dose, respectively. For the head and neck tumors, the mean doses to the spinal cord and parotid glands were 47% and 49%, respectively. For the pelvic tumors, the mean dose to the bladder, rectum, and small bowel were 51%, 63%, and 22%, respectively. Local failure occurred in one patient. IMRT resulted in substantial sparing of surrounding critical structures and acceptable local control rate for these tumors in children. Further follow-up is needed to assess long-term local control and late effects. PMID- 17121436 TI - Design and development of motorized multileaf collimator for telecobalt unit. AB - A manual multileaf collimator developed for telecobalt unit was motorized to accomplish the easy movement of the leaves. The required field shaping using MLC could be achieved by either using template or display. The beam characteristics were investigated and then compared with those of customized blocks. The maximum interleaf leakage and the percentage of transmission measured at the depth of maximum ionization (0.5cm) were found to be 2.7% and 2.4%, respectively. The field shaping performed by the MLC was verified using film dosimetry. The comparative study of treatment plans of 3DCRT and IMRT between (60)Co beam and 6 MV beams was carried out. This MLC could be used as a substitute for conventional blocks in static fields, there by eliminating the effort and cost of fabricating customized blocks, the need for storage space for blocks and other practical difficulties during the process of the block making. It is also demonstrated that if a provision for IMRT delivery with MLC for (60)Co is made, could be a cost effective alternative to IMRT with 6 MV beam. PMID- 17121437 TI - Spatially fractionated GRID radiation treatment of advanced neck disease associated with head and neck cancer. AB - Advanced nodal disease associated with head and neck cancer warrants aggressive, often multi-modality therapy to maximize local-regional control. The expansion of a novel treatment paradigm developed by our institution includes the addition of a single-fraction of high dose spatially-fractionated radiation (GRID) to a conventional course of treatment. Between 1995 and 2002 a series of 27 patients (median age 65) with bulky N2-3 disease were treated. Median nodal tumor size was 7 cm. Two groups of patients were evaluated. Group 1 (N=14) patients received a median neck dose 69 Gy (range 54-79 Gy) plus GRID treatment. Group 2 (N=13) patients received a median neck dose of 59 Gy (range 54-72 Gy) plus GRID treatment followed by planned neck dissection. Patients were evaluated for local regional control, pathological response, survival, and morbidity. Median time to follow-up for Group 1 was 10 months (range 3-44 months). Neck control was 93%. Disease specific survival was 50%. Morbidity was limited to soft-tissue related damage and was mild. Median time to follow-up for Group 2 was 38 months (range 5 116 months). Pathologic complete response rate was 85%. Overall neck control rate was 92%. Disease specific survival was 85%. Surgical morbidity was limited to three wound healing complications. GRID treatment may be safely added to conventional treatment management of locally advanced neck disease related to cancer with acceptable morbidity. It may improve pathologic complete response rates in those patients who undergo planned neck dissection, possibly leading to improved survival. In patients with inoperable bulky disease, addition of GRID enhances local-regional control. PMID- 17121438 TI - Killing two birds with one stone: a dosimetric study of dual target radiosurgery using a single isocenter. AB - The treatment of hematogenous brain metastases is a frequent indication for stereotactic radiosurgery (SRS). It is common for more than one metastasis to be treated during the same SRS session. We retrospectively identified four cases where our m3 micro multileaf collimator (mMLC) was used to create two distinct apertures and treat adjacent lesions using a single isocenter. For these four cases, single isocenter plans with static conformal beams were dosimetrically compared to plans utilizing two isocenters with static conformal beams or conformal arcs. The effects on dose homogeneity, dose conformity, and the minimum isodose separating the two targets are minor and variable. On the other hand, the use of a single isocenter technique consistently halves delivery time and decreases the integral dose to normal tissue. For small adjacent metastases, which can simultaneously be encompassed within the high-resolution portion of the m3/Novalis mMLC collimator, the use of a single rather than a dual isocenter technique is feasible and generally advantageous. PMID- 17121439 TI - High intensity focused ultrasound for prostate cancer: a review of the scientific foundation, technology and clinical outcomes. AB - High Intensity Focused Ultrasound (HIFU) is a definitive treatment for localized prostate cancer that is currently utilized most in Europe and Japan but it not yet approved by the FDA for this indication. Within the armamentarium of definitive prostate cancer therapies it is unique as it is truly non-invasive and does not involve incision or excision. The purpose of this paper is to review the scientific foundation of the technology as well as the clinical outcomes of commercially available devices. The scientific foundation of HIFU is reviewed in terms of how it has resulted in the development of commercially available equipment. MEDLINE was used to search the medical literature for publications pertaining to HIFU for prostate cancer as a primary therapy in terms of clinical outcomes. Biochemical disease free rates as well as negative biopsy rates are reviewed. Different engineering optimization strategies in the face of technicalities inherent to HIFU for prostate cancer have led to the development of two distinct commercially available devices. Each has their own merits and limitations. HIFU provides excellent biochemical and local control and results appear to be durable. Clinical outcomes are similar for the two technologies developed but are difficult to compare due to different lengths of follow-up and varying patient populations. HIFU is a technically advanced definitive local therapy for prostate cancer. Short and medium term results are encouraging and its role as a primary therapy for prostate cancer continues to be defined as more results become available. PMID- 17121440 TI - Publishing an article in the journal of school nursing: an overview of the publication process. PMID- 17121441 TI - The healthy learner model for student chronic condition management--part I. AB - A significant number of children have chronic health conditions that interfere with normal activities, including school attendance and active participation in the learning process. Management of students' chronic conditions is complex and requires an integrated system. Models to improve chronic disease management have been developed for the medical system and public health. Programs that address specific chronic disease management or coordinate school health services have been implemented in schools. Lacking is a comprehensive, integrated model that links schools, students, parents, health care, and other community providers. The Healthy Learner Model for chronic condition management identifies seven elements for creating, implementing, and sustaining an efficient and effective, comprehensive community-based system for improving the management of chronic conditions for school children. It has provided the framework for successful chronic condition management in an urban school district and is proposed for replication in other districts and communities. PMID- 17121442 TI - The healthy learner model for student chronic condition management--part II: the asthma initiative. AB - The Healthy Learner Asthma Initiative (HLAI) was designed as a comprehensive, school-community initiative to improve asthma management and produce healthy learners. National asthma guidelines were translated into components of asthma management in the school setting that defined performance expectations and lead to greater quality and consistency of asthma care. The HLAI incorporated evidence based practice and introduced the role of the asthma resource nurse. Leadership, capacity building, and strong partnerships among school nurses, students, families, and health care providers were essential to the implementation and sustainability of the HLAI. Professional school nursing and evaluation were defined as key requisites to a successful initiative. Evaluation results indicated positive effects on nursing practice, fewer asthma visits to the health office, and better attendance among students who received asthma care in the school health office. The HLAI provided the basis for development of the Healthy Learner Model for Student Chronic Condition Management. PMID- 17121443 TI - The cognitive and academic impact of sickle cell disease. AB - Sickle cell disease (SCD) affects over 30,000 students in the United States. Central nervous system complications are widespread among students with SCD and include stroke, silent cerebral infarction, and cognitive impairment. The effects of these complications may lead to academic failure, limited career options, and for some, total disability. Despite studies describing the significant academic and cognitive impact of sickle cell disease, reports describing interventions are limited. There is a lack of awareness among educators of the academic risks associated with sickle cell disease and a lack of appropriate resource allocation. The school nurse, as community health advocate, will be called upon to bridge the gap among healthcare providers, parents, students, and educators. This article provides a review of both recent and landmark studies describing the cognitive and academic impact of sickle cell disease and discusses the role of the school nurse as an advocate, liaison, and educator. PMID- 17121444 TI - The experiences of school nurses caring for students receiving continuous subcutaneous insulin infusion therapy. AB - Diabetes mellitus is the most common metabolic disorder in childhood. Today, children with diabetes are receiving new technologically advanced treatment options, such as continuous subcutaneous insulin infusion (CSII) therapy. School nurses are the primary health caregivers of children with diabetes during school hours. Therefore, it is important to determine their perceptions, resources, and resource needs when caring for students with continuous subcutaneous insulin infusion or insulin pump therapy. This study uses a phenomenological approach to examine the experiences of school nurses caring for students receiving insulin pump therapy. Eleven school nurses were interviewed using semistructured taped interviews. The nurses' responses indicated that they were "scared" when first caring for students with continuous subcutaneous insulin infusion therapy. However, they were able to work through their fear by using their resources and gaining more knowledge and hands-on experience with insulin pumps. The data also revealed that school nurses who were able to learn the language of continuous subcutaneous insulin infusion therapy and successfully deal with pump problems developed trusting and knowing relationships with students, teachers, and parents. PMID- 17121445 TI - Childhood nutrition: perceptions of caretakers in a low-income urban setting. AB - The incidence of overweight and obese children, especially those from low-income and minority backgrounds, continues to rise. Multiple factors contribute to the rising rates. In order to gain an understanding of factors contributing to obesity in low-income families, a qualitative study was conducted with the purpose of gaining knowledge of low-income urban caretakers' understanding and attitudes regarding children's nutrition. A focused ethnography was used as a means of understanding behavior within the context of a person's cultural environment. The sample was 17 caretakers of children in the 1st-3rd grades. Four focus groups were conducted. Two themes emerged from caretakers' perceptions: knowing the right things children should eat and balancing healthy nutrition with unhealthy choices. Four categories emerged regarding influences on food choices: tradition, finances, time constraints, and role models. Lastly, five barriers and three facilitating factors emerged. Implications of the study findings for school nurses include the need, when implementing healthy eating programs for school children, to gain information from caretakers about their perceptions of childhood nutrition. PMID- 17121446 TI - Knowledge of school nurses in the U.K. regarding sexual health education. AB - In the U.K., a current government health strategy indicates that school nurses should be key contributors to sexual health education because they have access to the school-age population. However, there appears to be little research that investigates whether school nurses are the most appropriate health care professionals or indeed have sufficient knowledge to contribute to this topic in the schools. The purpose of this study was to examine the sexual health knowledge of school nurses, using a survey to assess school nurses employed in a large central region of the U.K. The results suggest that although their general knowledge of the topic was adequate, school nurses have insufficient knowledge to effectively teach about sexually transmitted infections or emergency contraception. This has implications for school nursing because it appears that school nurses may have inadequate knowledge about sexual health to contribute to sex education in schools. PMID- 17121449 TI - Adherence to HAART: a systematic review of developed and developing nation patient-reported barriers and facilitators. AB - BACKGROUND: Adherence to highly active antiretroviral therapy (HAART) medication is the greatest patient-enabled predictor of treatment success and mortality for those who have access to drugs. We systematically reviewed the literature to determine patient-reported barriers and facilitators to adhering to antiretroviral therapy. METHODS AND FINDINGS: We examined both developed and developing nations. We searched the following databases: AMED (inception to June 2005), Campbell Collaboration (inception to June 2005), CinAhl (inception to June 2005), Cochrane Library (inception to June 2005), Embase (inception to June 2005), ERIC (inception to June 2005), MedLine (inception to June 2005), and NHS EED (inception to June 2005). We retrieved studies conducted in both developed and developing nation settings that examined barriers and facilitators addressing adherence. Both qualitative and quantitative studies were included. We independently, in duplicate, extracted data reported in qualitative studies addressing adherence. We then examined all quantitative studies addressing barriers and facilitators noted from the qualitative studies. In order to place the findings of the qualitative studies in a generalizable context, we meta analyzed the surveys to determine a best estimate of the overall prevalence of issues. We included 37 qualitative studies and 47 studies using a quantitative methodology (surveys). Seventy-two studies (35 qualitative) were conducted in developed nations, while the remaining 12 (two qualitative) were conducted in developing nations. Important barriers reported in both economic settings included fear of disclosure, concomitant substance abuse, forgetfulness, suspicions of treatment, regimens that are too complicated, number of pills required, decreased quality of life, work and family responsibilities, falling asleep, and access to medication. Important facilitators reported by patients in developed nation settings included having a sense of self-worth, seeing positive effects of antiretrovirals, accepting their seropositivity, understanding the need for strict adherence, making use of reminder tools, and having a simple regimen. Among 37 separate meta-analyses examining the generalizability of these findings, we found large heterogeneity. CONCLUSIONS: We found that important barriers to adherence are consistent across multiple settings and countries. Research is urgently needed to determine patient-important factors for adherence in developing world settings. Clinicians should use this information to engage in open discussion with patients to promote adherence and identify barriers and facilitators within their own populations. PMID- 17121450 TI - Complement lysis activity in autologous plasma is associated with lower viral loads during the acute phase of HIV-1 infection. AB - BACKGROUND: To explore the possibility that antibody-mediated complement lysis contributes to viremia control in HIV-1 infection, we measured the activity of patient plasma in mediating complement lysis of autologous primary virus. METHODS AND FINDINGS: Sera from two groups of patients-25 with acute HIV-1 infection and 31 with chronic infection-were used in this study. We developed a novel real-time PCR-based assay strategy that allows reliable and sensitive quantification of virus lysis by complement. Plasma derived at the time of virus isolation induced complement lysis of the autologous virus isolate in the majority of patients. Overall lysis activity against the autologous virus and the heterologous primary virus strain JR-FL was higher at chronic disease stages than during the acute phase. Most strikingly, we found that plasma virus load levels during the acute but not the chronic infection phase correlated inversely with the autologous complement lysis activity. Antibody reactivity to the envelope (Env) proteins gp120 and gp41 were positively correlated with the lysis activity against JR-FL, indicating that anti-Env responses mediated complement lysis. Neutralization and complement lysis activity against autologous viruses were not associated, suggesting that complement lysis is predominantly caused by non-neutralizing antibodies. CONCLUSIONS: Collectively our data provide evidence that antibody mediated complement virion lysis develops rapidly and is effective early in the course of infection; thus it should be considered a parameter that, in concert with other immune functions, steers viremia control in vivo. PMID- 17121451 TI - What is the best approach to reducing birth defects associated with isotretinoin? AB - BACKGROUND TO THE DEBATE: Isotretinoin is an effective treatment for severe acne, a condition which can be physically, emotionally, and socially disabling. Because the drug is teratogenic, causing severe birth defects, women taking the drug are directed to avoid pregnancy. In the United States, a series of risk reduction programs have been implemented that aim to prevent pregnant women from taking the drug and to prevent women taking it from getting pregnant. The most recent, and most stringent, is an Internet-based, performance-linked system called iPLEDGE, which tries to ensure that the drug is dispensed only when there is documentary proof that the patient is not pregnant and is using two forms of birth control. Is iPLEDGE the best way to reduce isotretinoin birth defects, or is it an unproven and overly burdensome system? PMID- 17121452 TI - Six imprisoned health-care workers in Libya are pawns in a far larger strategic game. The repercussions are enormous. PMID- 17121453 TI - Interdigitated paralemniscal and lemniscal pathways in the mouse barrel cortex. AB - Primary sensory cortical areas receive information through multiple thalamic channels. In the rodent whisker system, lemniscal and paralemniscal thalamocortical projections, from the ventral posteromedial nucleus (VPM) and posterior medial nucleus (POm) respectively, carry distinct types of sensory information to cortex. Little is known about how these separate streams of activity are parsed and integrated within the neocortical microcircuit. We used quantitative laser scanning photostimulation to probe the organization of functional thalamocortical and ascending intracortical projections in the mouse barrel cortex. To map the thalamocortical projections, we recorded from neocortical excitatory neurons while stimulating VPM or POm. Neurons in layers (L)4, L5, and L6A received dense input from thalamus (L4, L5B, and L6A from VPM; and L5A from POm), whereas L2/3 neurons rarely received thalamic input. We further mapped the lemniscal and paralemniscal circuits from L4 and L5A to L2/3. Lemniscal L4 neurons targeted L3 within a column. Paralemniscal L5A neurons targeted a superficial band (thickness, 60 mum) of neurons immediately below L1, defining a functionally distinct L2 in the mouse barrel cortex. L2 neurons received input from lemniscal L3 cells and paralemniscal L5A cells spread over multiple columns. Our data indicate that lemniscal and paralemniscal information is segregated into interdigitated cortical layers. PMID- 17121454 TI - A posterior centre establishes and maintains polarity of the Caenorhabditis elegans embryo by a Wnt-dependent relay mechanism. AB - Cellular polarity is a general feature of animal development. However, the mechanisms that establish and maintain polarity in a field of cells or even in the whole embryo remain elusive. Here we provide evidence that in the Caenorhabditis elegans embryo, the descendants of P1, the posterior blastomere of the 2-cell stage, constitute a polarising centre that orients the cell divisions of most of the embryo. This polarisation depends on a MOM-2/Wnt signal originating from the P1 descendants. Furthermore, we show that the MOM-2/Wnt signal is transduced from cell to cell by a relay mechanism. Our findings suggest how polarity is first established and then maintained in a field of cells. According to this model, the relay mechanism constantly orients the polarity of all cells towards the polarising centre, thus organising the whole embryo. This model may also apply to other systems such as Drosophila and vertebrates. PMID- 17121455 TI - Functional convergence of neurons generated in the developing and adult hippocampus. AB - The dentate gyrus of the hippocampus contains neural progenitor cells (NPCs) that generate neurons throughout life. Developing neurons of the adult hippocampus have been described in depth. However, little is known about their functional properties as they become fully mature dentate granule cells (DGCs). To compare mature DGCs generated during development and adulthood, NPCs were labeled at both time points using retroviruses expressing different fluorescent proteins. Sequential electrophysiological recordings from neighboring neurons of different ages were carried out to quantitatively study their major synaptic inputs: excitatory projections from the entorhinal cortex and inhibitory afferents from local interneurons. Our results show that DGCs generated in the developing and adult hippocampus display a remarkably similar afferent connectivity with regard to both glutamate and GABA, the major neurotransmitters. We also demonstrate that adult-born neurons can fire action potentials in response to an excitatory drive, exhibiting a firing behavior comparable to that of neurons generated during development. We propose that neurons born in the developing and adult hippocampus constitute a functionally homogeneous neuronal population. These observations are critical to understanding the role of adult neurogenesis in hippocampal function. PMID- 17121457 TI - Ten simple rules for selecting a postdoctoral position. PMID- 17121456 TI - Iron regulation of the major virulence factors in the AIDS-associated pathogen Cryptococcus neoformans. AB - Iron overload is known to exacerbate many infectious diseases, and conversely, iron withholding is an important defense strategy for mammalian hosts. Iron is a critical cue for Cryptococcus neoformans because the fungus senses iron to regulate elaboration of the polysaccharide capsule that is the major virulence factor during infection. Excess iron exacerbates experimental cryptococcosis and the prevalence of this disease in Sub-Saharan Africa has been associated with nutritional and genetic aspects of iron loading in the background of the HIV/AIDS epidemic. We demonstrate that the iron-responsive transcription factor Cir1 in Cr. neoformans controls the regulon of genes for iron acquisition such that cir1 mutants are "blind" to changes in external iron levels. Cir1 also controls the known major virulence factors of the pathogen including the capsule, the formation of the anti-oxidant melanin in the cell wall, and the ability to grow at host body temperature. Thus, the fungus is remarkably tuned to perceive iron as part of the disease process, as confirmed by the avirulence of the cir1 mutant; this characteristic of the pathogen may provide opportunities for antifungal treatment. PMID- 17121458 TI - Meta-analysis of differentiating mouse embryonic stem cell gene expression kinetics reveals early change of a small gene set. AB - Stem cell differentiation involves critical changes in gene expression. Identification of these should provide endpoints useful for optimizing stem cell propagation as well as potential clues about mechanisms governing stem cell maintenance. Here we describe the results of a new meta-analysis methodology applied to multiple gene expression datasets from three mouse embryonic stem cell (ESC) lines obtained at specific time points during the course of their differentiation into various lineages. We developed methods to identify genes with expression changes that correlated with the altered frequency of functionally defined, undifferentiated ESC in culture. In each dataset, we computed a novel statistical confidence measure for every gene which captured the certainty that a particular gene exhibited an expression pattern of interest within that dataset. This permitted a joint analysis of the datasets, despite the different experimental designs. Using a ranking scheme that favored genes exhibiting patterns of interest, we focused on the top 88 genes whose expression was consistently changed when ESC were induced to differentiate. Seven of these (103728_at, 8430410A17Rik, Klf2, Nr0b1, Sox2, Tcl1, and Zfp42) showed a rapid decrease in expression concurrent with a decrease in frequency of undifferentiated cells and remained predictive when evaluated in additional maintenance and differentiating protocols. Through a novel meta-analysis, this study identifies a small set of genes whose expression is useful for identifying changes in stem cell frequencies in cultures of mouse ESC. The methods and findings have broader applicability to understanding the regulation of self renewal of other stem cell types. PMID- 17121459 TI - Robustness and fragility in immunosenescence. AB - We construct a model to study tradeoffs associated with aging in the adaptive immune system, focusing on cumulative effects of replacing naive cells with memory cells. Binding affinities are characterized by a stochastic shape space model. System loss arising from an individual infection is associated with disease severity, as measured by the total antigen population over the course of an infection. We monitor evolution of cell populations on the shape space over a string of infections, and find that the distribution of losses becomes increasingly heavy-tailed with time. Initially this lowers the average loss: the memory cell population becomes tuned to the history of past exposures, reducing the loss of the system when subjected to a second, similar infection. This is accompanied by a corresponding increase in vulnerability to novel infections, which ultimately causes the expected loss to increase due to overspecialization, leading to increasing fragility with age (i.e., immunosenescence). In our model, immunosenescence is not the result of a performance degradation of some specific lymphocyte, but rather a natural consequence of the built-in mechanisms for system adaptation. This "robust, yet fragile" behavior is a key signature of Highly Optimized Tolerance. PMID- 17121460 TI - Computational inference of neural information flow networks. AB - Determining how information flows along anatomical brain pathways is a fundamental requirement for understanding how animals perceive their environments, learn, and behave. Attempts to reveal such neural information flow have been made using linear computational methods, but neural interactions are known to be nonlinear. Here, we demonstrate that a dynamic Bayesian network (DBN) inference algorithm we originally developed to infer nonlinear transcriptional regulatory networks from gene expression data collected with microarrays is also successful at inferring nonlinear neural information flow networks from electrophysiology data collected with microelectrode arrays. The inferred networks we recover from the songbird auditory pathway are correctly restricted to a subset of known anatomical paths, are consistent with timing of the system, and reveal both the importance of reciprocal feedback in auditory processing and greater information flow to higher-order auditory areas when birds hear natural as opposed to synthetic sounds. A linear method applied to the same data incorrectly produces networks with information flow to non-neural tissue and over paths known not to exist. To our knowledge, this study represents the first biologically validated demonstration of an algorithm to successfully infer neural information flow networks. PMID- 17121461 TI - SabA is the H. pylori hemagglutinin and is polymorphic in binding to sialylated glycans. AB - Adherence of Helicobacter pylori to inflamed gastric mucosa is dependent on the sialic acid-binding adhesin (SabA) and cognate sialylated/fucosylated glycans on the host cell surface. By in situ hybridization, H. pylori bacteria were observed in close association with erythrocytes in capillaries and post-capillary venules of the lamina propria of gastric mucosa in both infected humans and Rhesus monkeys. In vivo adherence of H. pylori to erythrocytes may require molecular mechanisms similar to the sialic acid-dependent in vitro agglutination of erythrocytes (i.e., sialic acid-dependent hemagglutination). In this context, the SabA adhesin was identified as the sialic acid-dependent hemagglutinin based on sialidase-sensitive hemagglutination, binding assays with sialylated glycoconjugates, and analysis of a series of isogenic sabA deletion mutants. The topographic presentation of binding sites for SabA on the erythrocyte membrane was mapped to gangliosides with extended core chains. However, receptor mapping revealed that the NeuAcalpha2-3Gal-disaccharide constitutes the minimal sialylated binding epitope required for SabA binding. Furthermore, clinical isolates demonstrated polymorphism in sialyl binding and complementation analysis of sabA mutants demonstrated that polymorphism in sialyl binding is an inherent property of the SabA protein itself. Gastric inflammation is associated with periodic changes in the composition of mucosal sialylation patterns. We suggest that dynamic adaptation in sialyl-binding properties during persistent infection specializes H. pylori both for individual variation in mucosal glycosylation and tropism for local areas of inflamed and/or dysplastic tissue. PMID- 17121462 TI - A translocated bacterial protein protects vascular endothelial cells from apoptosis. AB - The modulation of host cell apoptosis by bacterial pathogens is of critical importance for the outcome of the infection process. The capacity of Bartonella henselae and B. quintana to cause vascular tumor formation in immunocompromised patients is linked to the inhibition of vascular endothelial cell (EC) apoptosis. Here, we show that translocation of BepA, a type IV secretion (T4S) substrate, is necessary and sufficient to inhibit EC apoptosis. Ectopic expression in ECs allowed mapping of the anti-apoptotic activity of BepA to the Bep intracellular delivery domain, which, as part of the signal for T4S, is conserved in other T4S substrates. The anti-apoptotic activity appeared to be limited to BepA orthologs of B. henselae and B. quintana and correlated with (i) protein localization to the host cell plasma membrane, (ii) elevated levels of intracellular cyclic adenosine monophosphate (cAMP), and (iii) increased expression of cAMP-responsive genes. The pharmacological elevation of cAMP levels protected ECs from apoptosis, indicating that BepA mediates anti-apoptosis by heightening cAMP levels by a plasma membrane-associated mechanism. Finally, we demonstrate that BepA mediates protection of ECs against apoptosis triggered by cytotoxic T lymphocytes, suggesting a physiological context in which the anti-apoptotic activity of BepA contributes to tumor formation in the chronically infected vascular endothelium. PMID- 17121463 TI - Anthrax, but not Bacillus anthracis? PMID- 17121464 TI - Dissecting the WRKY web of plant defense regulators. PMID- 17121465 TI - Genomic imprinting in mammals: emerging themes and established theories. AB - The epigenetic events that occur during the development of the mammalian embryo are essential for correct gene expression and cell-lineage determination. Imprinted genes are expressed from only one parental allele due to differential epigenetic marks that are established during gametogenesis. Several theories have been proposed to explain the role that genomic imprinting has played over the course of mammalian evolution, but at present it is not clear if a single hypothesis can fully account for the diversity of roles that imprinted genes play. In this review, we discuss efforts to define the extent of imprinting in the mouse genome, and suggest that different imprinted loci may have been wrought by distinct evolutionary forces. We focus on a group of small imprinted domains, which consist of paternally expressed genes embedded within introns of multiexonic transcripts, to discuss the evolution of imprinting at these loci. PMID- 17121466 TI - Inference of splicing regulatory activities by sequence neighborhood analysis. AB - Sequence-specific recognition of nucleic-acid motifs is critical to many cellular processes. We have developed a new and general method called Neighborhood Inference (NI) that predicts sequences with activity in regulating a biochemical process based on the local density of known sites in sequence space. Applied to the problem of RNA splicing regulation, NI was used to predict hundreds of new exonic splicing enhancer (ESE) and silencer (ESS) hexanucleotides from known human ESEs and ESSs. These predictions were supported by cross-validation analysis, by analysis of published splicing regulatory activity data, by sequence conservation analysis, and by measurement of the splicing regulatory activity of 24 novel predicted ESEs, ESSs, and neutral sequences using an in vivo splicing reporter assay. These results demonstrate the ability of NI to accurately predict splicing regulatory activity and show that the scope of exonic splicing regulatory elements is substantially larger than previously anticipated. Analysis of orthologous exons in four mammals showed that the NI score of ESEs, a measure of function, is much more highly conserved above background than ESE primary sequence. This observation indicates a high degree of selection for ESE activity in mammalian exons, with surprisingly frequent interchangeability between ESE sequences. PMID- 17121467 TI - Pigment pattern in jaguar/obelix zebrafish is caused by a Kir7.1 mutation: implications for the regulation of melanosome movement. AB - Many animals have a variety of pigment patterns, even within a species, and these patterns may be one of the driving forces of speciation. Recent molecular genetic studies on zebrafish have revealed that interaction among pigment cells plays a key role in pattern formation, but the mechanism of pattern formation is unclear. The zebrafish jaguar/obelix mutant has broader stripes than wild-type fish. In this mutant, the development of pigment cells is normal but their distribution is altered, making these fish ideal for studying the process of pigment pattern formation. Here, we utilized a positional cloning method to determine that the inwardly rectifying potassium channel 7.1 (Kir7.1) gene is responsible for pigment cell distribution among jaguar/obelix mutant fish. Furthermore, in jaguar/obelix mutant alleles, we identified amino acid changes in the conserved region of Kir7.1, each of which affected K(+) channel activity as demonstrated by patch-clamp experiments. Injection of a bacterial artificial chromosome containing the wild-type Kir7.1 genomic sequence rescued the jaguar/obelix phenotype. From these results, we conclude that mutations in Kir7.1 are responsible for jaguar/obelix. We also determined that the ion channel function defect of melanophores expressing mutant Kir7.1 altered the cellular response to external signals. We discovered that mutant melanophores cannot respond correctly to the melanosome dispersion signal derived from the sympathetic neuron and that melanosome aggregation is constitutively activated. In zebrafish and medaka, it is well known that melanosome aggregation and subsequent melanophore death increase when fish are kept under constant light conditions. These observations indicate that melanophores of jaguar/obelix mutant fish have a defect in the signaling pathway downstream of the alpha2-adrenoceptor. Taken together, our results suggest that the cellular defect of the Kir7.1 mutation is directly responsible for the pattern change in the jaguar/obelix mutant. PMID- 17121468 TI - Operon conservation and the evolution of trans-splicing in the phylum Nematoda. AB - The nematode Caenorhabditis elegans is unique among model animals in that many of its genes are cotranscribed as polycistronic pre-mRNAs from operons. The mechanism by which these operonic transcripts are resolved into mature mRNAs includes trans-splicing to a family of SL2-like spliced leader exons. SL2-like spliced leaders are distinct from SL1, the major spliced leader in C. elegans and other nematode species. We surveyed five additional nematode species, representing three of the five major clades of the phylum Nematoda, for the presence of operons and the use of trans-spliced leaders in resolution of polycistronic pre-mRNAs. Conserved operons were found in Pristionchus pacificus, Nippostrongylus brasiliensis, Strongyloides ratti, Brugia malayi, and Ascaris suum. In nematodes closely related to the rhabditine C. elegans, a related family of SL2-like spliced leaders is used for operonic transcript resolution. However, in the tylenchine S. ratti operonic transcripts are resolved using a family of spliced leaders related to SL1. Non-operonic genes in S. ratti may also receive these SL1 variants. In the spirurine nematodes B. malayi and A. suum operonic transcripts are resolved using SL1. Mapping these phenotypes onto the robust molecular phylogeny for the Nematoda suggests that operons evolved before SL2 like spliced leaders, which are an evolutionary invention of the rhabditine lineage. PMID- 17121469 TI - Opposite root growth phenotypes of hy5 versus hy5 hyh mutants correlate with increased constitutive auxin signaling. AB - The Arabidopsis transcription factor HY5 controls light-induced gene expression downstream of photoreceptors and plays an important role in the switch of seedling shoots from dark-adapted to light-adapted development. In addition, HY5 has been implicated in plant hormone signaling, accounting for the accelerated root system growth phenotype of hy5 mutants. Mutants in the close HY5 homolog HYH resemble wild-type, despite the largely similar expression patterns and levels of HY5 and HYH, and the functional equivalence of the respective proteins. Moreover, the relative contribution of HYH to the overall activity of the gene pair is increased by an alternative HYH transcript, which encodes a stabilized protein. Consistent with the enhanced root system growth observed in hy5 loss-of-function mutants, constitutively overexpressed alternative HYH inhibits root system growth. Paradoxically, however, in double mutants carrying hy5 and hyh null alleles, the hy5 root growth phenotype is suppressed rather than enhanced. Even more surprisingly, compared to wild-type, root system growth is diminished in hy5 hyh double mutants. In addition, the double mutants display novel shoot phenotypes that are absent from either single mutant. These include cotyledon fusions and defective vasculature, which are typical for mutants in genes involved in the transcriptional response to the plant hormone auxin. Indeed, many auxin-responsive and auxin signaling genes are misexpressed in hy5 mutants, and at a higher number and magnitude in hy5 hyh mutants. Therefore, auxin-induced transcription is constitutively activated at different levels in the two mutant backgrounds. Our data support the hypothesis that the opposite root system phenotypes of hy5 single and hy5 hyh double mutants represent the morphological response to a quantitative gradient in the same molecular process, that is gradually increased constitutive auxin signaling. The data also suggest that HY5 and HYH are important negative regulators of auxin signaling amplitude in embryogenesis and seedling development. PMID- 17121470 TI - Alternative splicing at NAGNAG acceptors: simply noise or noise and more? PMID- 17121472 TI - Speak v. pro-test--the Newsnight animal experimentation debate--round 1. PMID- 17121473 TI - The comparative effects of metals on the hatching of earthworm cocoons. AB - To establish the use of Metaphire posthuma as a sensitive model for ecotoxicological studies, the comparative effects of five metals on the hatching profiles of the cocoons of the earthworms, Metaphire posthuma, Eisenia foetida and Perionyx excavatus, were studied. The cocoons of the three species of earthworms were exposed to copper, chromium (III), chromium (VI), lead and zinc at 1.25, 2.5, 5.0 and 10.0 ppm. Viable cocoons were incubated at 20 +/- degrees C by using the immersion method. The results indicated that the inhibition of cocoon hatching was concentration dependent. The normal hatching, delayed hatching and non-viability of cocoons were recorded. At a concentration of 1.25 ppm, there was almost no effect on the hatching of the cocoons of all three species of earthworms, except when exposed to chromium (VI), but higher concentrations (2.5 and 5.0 ppm) caused severe effects. It was concluded that M. posthuma was more sensitive than the other two species, and that it is a suitable model for use in ecotoxicity testing. PMID- 17121474 TI - Comparison of bovine udder skin with human and porcine skin in percutaneous permeation experiments. AB - Rat and pig animal skin has been the most common replacement material for human skin for use in in vitro permeability experiments. Unfortunately, the permeability barrier of skin from laboratory animals is known to be relatively weak, due to significant follicular transport. Pig skin has been shown to be a suitable model for human skin. Unfortunately, it cannot be gathered from the regular slaughtering process, which makes it unsuitable for permeation experiments. We therefore studied the suitability of bovine udder skin, an untreated waste material of the butchering process, as a possible replacement material for use in in vitro permeability tests. We investigated the barrier strength of bovine udder skin against four different substances, and its histology and lipid profile, in comparison with pig skin and heat separated human epidermis. Pig and human skin were found to be equally permeable, whilst bovine udder skin seemed to exhibit a weaker, but less variable, barrier against caffeine, benzoic acid, testosterone, and flufenamic acid. The skin of all three species contained variable contents of the major lipid classes: cholesterol, ceramides, cholesterol ester, fatty acids and triglycerides. Morphological differences mainly comprised variations in the density of hair follicles. Based on these results, the amount of free fatty acids and triglycerides and the density of hair follicles seem to be important factors in the differences between the skin barriers in the three species. PMID- 17121475 TI - Comparison of impedance-based sensors for cell adhesion monitoring and in vitro methods for detecting cytotoxicity induced by chemicals. AB - The recent European Commission REACH (Registration, Evaluation and Authorisation of Chemicals) policy outlines a plan for toxicological testing by using alternative non-animal in vitro methods. In this context, there is a need to develop and standardise high-throughput screening (HTS) methods for studying the cytotoxicity induced by chemicals. Electrochemical impedance spectroscopy (EIS) can be considered as a complementary technique to alternative in vitro testing for studying cell adhesion to the substrate, and can give real-time and kinetic information on cell responses to a toxicant. This paper describes the development of a home-made chip based on impedance spectroscopy, and its application in studying the kinetics of BALB/3T3 cell adhesion and the cellular responses to a toxic product as a function of time. Concentrations of sodium arsenite, ranging from 10 microM up to 1000 microM, were tested in the system, and the results were compared with those obtained with standard protocols used to study basal cytotoxicity induced by chemicals in the BALB/3T3 cell line. The results show that the sensitivity of the developed chip was better than that with the MTT test, with the additional advantages of online monitoring. PMID- 17121476 TI - The practical application of three validated in vitro embryotoxicity tests. The report and recommendations of an ECVAM/ZEBET workshop (ECVAM workshop 57). PMID- 17121477 TI - The UK Food Standards Agency draft report on variability and uncertainty in toxicology: a response by FRAME. AB - At the request of the Food Standards Agency, the Committee on Toxicity of Chemicals in Food, Consumer Products and the Environment (COT) established a Working Group on Variation and Uncertainty in Toxicology (WG VUT). In April 2006, the WG VUT produced a draft report for public consultation. FRAME made a submission in response to this consultation in July 2006. We commend the WG VUT for its comprehensive account of many of the problems associated with risk assessment, and for making recommendations about the problems that need to be addressed. We were particularly encouraged by the WG VUTs recognition of the need for guidelines on how toxicological studies should be conducted and data analysed. However, we believe that the report has not achieved all of its objectives. It does not adequately consider how modern technologies, experimental design, statistical analysis and species extrapolation can be used in practice to address variability and uncertainty. There is a disproportionate focus on the sources of variability and uncertainty in human data, with relatively little consideration of how variation and uncertainty due to animal tests can be addressed. Furthermore, it is clear that, until the advantages and limitations of all toxicological methods are fully appraised and testing strategies and guidelines are agreed, the scope for improving the existing approaches to risk assessment will be severely limited. Hence, the use of alternative methods for hazard identification and characterisation merit more consideration than they were given in the draft report. PMID- 17121482 TI - Rheumatology in 2006: crossroads or crisis? AB - Rheumatology has made remarkable advances in patient treatment in the past decade related to the impressive array of new drugs that have been approved or are undergoing clinical trial. While this situation should engender optimism for the future, concerns about sustaining momentum have been raised. These concerns relate to uncertainty in the research agenda for major diseases such as osteoarthritis and fibromyalgia, lack of informatics systems to allow accurate assessment of risks and benefits of new treatments, and a paucity of clinical trials in rheumatoid arthritis aimed at sustained remission or cure. Fortunately, the opportunities for the future remain very bright because of burgeoning research in biomedicine and outcomes assessment as well as progress in developing personalized medicine to individualize treatment better. PMID- 17121483 TI - The pathogenesis of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is due to a combination of phagocytosis and anaphylaxis gone awry. Immune complexes create their havoc via Fc gamma III receptors that signal via the tyrosine phosphorylation immunoreceptor pathway. Anaphylatoxins, mainly C5a, signal via the MEK kinase cascade, and both engage in cross-talk via NF kappa B. Aspirin-III blocks signals sent by both: immune complexes via FCR, and anaphylatoxin via C5 receptors. Drugs that affect both pathways, for example anti-B cell monoclonal antibodies, such as rituximab, and anti-C5 monoclonal antibodies, such as pexelizumab are currently being investigated. We now appreciate that cytokines are important mediators of inflammation in RA but are downstream of the primary insults: immune complexes and anaphylatoxins. We can therefore begin to ask what the antigen or antigens might be that produce IgGs reactive to 7 S or 19 S rheumatoid factors. The primary antigens of RA could well be CPs, formed perhaps by oral bacteria. Once immunoglobulins become recognized by rheumatoid factors, immune complexes form, and these activate anaphylatoxins. Phagocytosis and anaphylaxis are the proximal events of RA. PMID- 17121484 TI - Pharmacogenetics in the rheumatic diseases. AB - Designing a therapeutic plan that involves the least risk of toxicity and the greatest chance of success is the goal of the modern physician. To better achieve this goal an understanding of the genetic basis for drug efficacy and toxicity is essential. Here we review the available information on the pharmacogenetics of drugs commonly used to treat rheumatic diseases in the hope that the application of this information to the patient will contribute to more effective and safer therapies for rheumatoid arthritis, systemic lupus erythematosus, and other inflammatory diseases. PMID- 17121485 TI - The role of the synovial fibroblast in rheumatoid arthritis: cartilage destruction and the regulation of matrix metalloproteinases. AB - Rheumatoid arthritis (RA) is a complex multisystem disease, the hallmark of which is pannus, the abnormal proliferative synovial tissue that serves as both propagator of the immune response and as the engine of tissue damage. Conceptually, pannus may be divided into two compartments (Fig. 1). The first, comprised by T cells, B cells, macrophages, and dendritic cells, is an immune compartment that exists in what was formerly the subintimal layer of normal synovium. These immune cells partake in antigen presentation, immunoglobulin production (including rheumatoid factor and anti-cyclic citrullinated protein [CCP] antibodies) and cytokine generation; the T cell is thought by many investigators to be the driving force coordinating these various activities. PMID- 17121486 TI - Psoriatic arthritis update. AB - Psoriatic arthritis is an inflammatory arthritis occurring in up to 30% of patients with psoriasis. Its clear distinction from rheumatoid arthritis has been described clinically, genetically, and immunohistologically. Updated classification criteria have been recently derived from a large international study. Key pathophysiologic cellular processes are being elucidated, increasing our understanding of potential targets of therapy. Therapies that target cells, such as activated T cells, and proinflammatory cytokines, such as tumor necrosis factor alpha (TNFalpha), are rational to pursue. Outcome measures have been "borrowed" from rheumatoid arthritis and psoriasis studies. A variety of domains are assessed including joints, skin, enthesium, dactylitis, spine, function, quality of life, and imaging assessment of disease activity and damage. The performance qualities of outcome measures in these various domains is being evaluated by the Group for Research and Assessment of Psoriasis and Psoriatic Arthritis (GRAPPA), and improved measures are being developed and validated specifically for psoriatic arthritis. Traditional therapies for psoriatic arthritis have included nonsteroidal anti-inflammatory agents, oral immunomodulatory drugs, topical creams, and light therapy. These therapies have been helpful in controlling both musculoskeletal and dermatologic aspects of the disease, but they may not be fully effective in all disease domains, may eventually show diminished benefit, and may produce treatment-limiting toxicities. In the past several years, use of biologic agents has generally yielded greater benefit across more domains, yielding significant and enduring benefits for clinical manifestations, function, and quality of life, and especially with the anti-TNF agents, inhibition of structural damage. Adverse effects with these agents can be significant but are usually manageable. Cost is also significant, but cost-effectiveness analysis is demonstrating reasonable trade-off between cost and benefit. PMID- 17121487 TI - Advantages and limitations of quantitative measures to assess rheumatoid arthritis: joint counts, radiographs, laboratory tests, and patient questionnaires. AB - Medical care is advanced by quantitative measures, all of which have advantages and limitations. No single "gold standard" measure, analogous to blood pressure, is available for diagnosis, prognosis, and management of rheumatoid arthritis (RA). Four types of measures have been used, including joint counts, radiographs, laboratory tests, and patient questionnaires. Joint counts are the most specific measure for RA but are poorly reproducible and not performed in most standard care. Radiographs provide an objective record of joint damage, but are scored quantitatively only in clinical research and have little prognostic value for long-term outcomes such as work disability and mortality. Laboratory tests are helpful when positive but frequently are "false negative"--for example, rheumatoid factor (RF), erythrocyte sedimentation rate, or C-reactive protein are normal in 30% to 45% of patients. "False positive" results are also seen; most people with RF or antinuclear antibody do not have a disease. Patient questionnaires are useful to assess and monitor patient status and provide the most significant predictive measures for long-term work disability and mortality. A multidimensional health assessment questionnaire is useful in all rheumatic diseases, with scoring templates and medical history information to save time for the rheumatologist and patient in standard care. PMID- 17121488 TI - Monitoring outcomes of arthritis and longitudinal data collection using patient questionnaires in routine care. AB - Though quantitative data might lead to improved information for clinical decisions, at the present time decisions in routine rheumatology practice generally are based largely on qualitative impressions, rather than on data. Patient questionnaires are readily accessible tools that the rheumatologist can use to go beyond impressions and to institute evidence-based guidelines appropriate to his or her own patient population and practice style. The Health Assessment Questionnaire (HAQ) and its derivatives have been shown to be the best predictors of functional and work disability, costs, joint replacement surgery, and mortality. Such questionnaires are at least as good as joint counts, radiographs, and laboratory tests in predicting these outcomes. Every encounter of a patient with a rheumatologist provides an opportunity to collect data. Based on experience with the Brooklyn Outcomes of Arthritis Registry Database, the author advocates distributing a waiting-room questionnaire to every patient who comes for an office visit. Potential benefits of recording questionnaire-based information include identifying trends or important changes in a patient's pain or physical function, providing a baseline for success with various treatment strategies for conditions of the rheumatologist's own practice, allowing patients an opportunity to express concerns, encouraging patients to disclose information they may feel is too minor to mention, and providing control data for research studies. A short questionnaire designed specifically for clinical, rather than research, use does not create a burden for office staff. Consistent use of patient questionnaires and systematic storage of the information gained can help document, track, and improve patient care in routine rheumatology practice. PMID- 17121489 TI - Epstein-Barr virus infection induces lupus autoimmunity. AB - Systemic lupus erythematosus (SLE or lupus) is a systemic autoimmune disease characterized by a constellation of varied clinical presentations, although the nearly universal presence of autoantibodies is a salient unifying feature. Ongoing research efforts focus on understanding the complex combination of genetic and environmental factors that lead to SLE in select individuals. Our previous work has demonstrated that years before diagnosis abnormal autoantibody responses are present in the sera of patients who will subsequently develop lupus and, further, that the initial targets of two of these key responses (anti-Sm B' and anti-60 kD Ro alone) have been identified for some patients. Indeed, our results suggest that the first lupus-specific autoantibodies arise from particular antibodies directed against Epstein-Barr virus Nuclear Antigen-1 (EBNA 1) and that infection with Epstein-Barr virus (EBV) is an environmental risk factor for lupus. The predicted sequence of events is normal immunity, followed by Epstein- Barr virus infection, the generation of anti-EBNA-1 antibodies, then followed by those particular anti-EBNA-1 antibodies that also bind lupus-specific autoantigens (Sm or Ro), followed by the development of more complex autoimmune responses, and, finally, culminating in clinical disease. Studies from others and those underway suggest that lupus patients have unusual immune responses to Epstein-Barr virus. In aggregate, these results are consistent with an immune response against Epstein-Barr virus being important in at least some patients for the initiation of lupus autoimmunity. PMID- 17121490 TI - Targeting of B cells in SLE: rationale and therapeutic opportunities. AB - B cells were first implicated in lupus pathogenesis because of their roles as autoantibody producers. B cells, which play important roles in (auto)immune recognition, are now understood to also have other functional capabilities that contribute to the recruitment and stimulation of T lymphocytes and cells of the innate immune system. Herein, these emerging insights are discussed as well as the newly recognized influence on B cells of the Toll-like receptors, which are postulated to be important sources of costimulation for autoreactive B cells. Also discussed is how B-cell survival factors may contribute to the loss of immune tolerance that leads to pathologic autoimmunity. These findings are part of the rationale for the development of new specific B-cell-targeted therapies. PMID- 17121491 TI - Update on antiphospholipid syndrome. AB - Revised criteria for the antiphospholipid syndrome were published in 2006. Major changes from the 1999 criteria are an increase in the time between two laboratory studies required for diagnosis from 6 to 12 weeks, the acceptance of antibody to beta2 glycoprotein I as a criterion, the exclusion of older age persons, and the acknowledgment of several associated findings such as livedo, heart valve disease, and antibody to prothrombin. New concepts of pathogenesis now invoke complement activation and participation of the innate immune system upstream to thrombosis. Warfarin remains the treatment of choice for patients who have suffered thrombosis, but antiplatelet agents and heparin are options. Target INR is 2.0-3.0. Treatment is potentially life-long, though options for withdrawal of treatment are under investigation. PMID- 17121492 TI - Treatment of ANCA-associated systemic vasculitis. AB - The antineutrophil cytoplasmic antibodies (ANCA)-associated small vessel vasculitides include Wegener's granulomatosis, Churg-Strauss syndrome, microscopic polyangiitis (MPA), and the renal limited form of MPA, also known as pauci-immune or idiopathic crescentic glomerulonephritis. ANCA are probably necessary but not sufficient for disease pathogenicity. Classical induction and maintenance therapy of these conditions with corticosteroids and long-term cyclophosphamide is associated with occasional relapse and major toxicities. Therefore, treatment regimens being investigated include induction with methotrexate or, especially for patients with more aggressive disease accompanied by renal insufficiency, therapies that include either pulses of methylprednisolone or plasma exchanges. Nontraditional options for maintenance therapy may include step-down treatment with azathioprine or mycophenolate mofetil. For patients with Wegener's granulomatosis, studies have shown a reduced occurrence of flares with the use of co-trimoxazole. Finally, although a carefully randomized controlled trial with etanercept demonstrated that this tumor necrosis factor (TNF)-blocking agent was not superior to conventional maintenance therapy, a biologic agent with a different mechanism of action, rituximab, may prove a satisfactory alternative. PMID- 17121493 TI - Osteoporosis: is there a rational approach to fracture prevention? AB - The most effective way to manage osteoporosis is to prevent fractures before they occur. To do this, a clinician needs to be aware of both the clinical risk factors that predispose a patient to an osteoporotic fracture and the patient's bone mineral density (BMD). An assessment of risk factors that increase fracture risk, including age, weight less than 125 pounds as an adult, family history of hip fracture, low-impact fractures as an adult, inability to rise up from a chair without using one's arms, presence of rheumatoid arthritis (RA), and use of glucocorticoid medication, in addition to low BMD, is necessary to assess fracture risk. Therefore, a complete history and BMD will improve the identification and treatment of patients at high risk of an osteoporotic fracture. Also, patients with systemic inflammatory diseases like RA or systemic lupus erythematosus have an increased risk of fracture owing to systemic inflammation independent of glucocorticoid use. These patients should be screened for osteoporotic risk factors, and BMD tests should be obtained. Treatment to prevent fractures should be initiated at a BMD (T score) <-1 to improve skeletal health in these patients. This review provides an update on the epidemiology of fractures, reviews fracture risk-factor assessment, and makes recommendations on how to screen patients and decide which patients would benefit from an intervention. Lastly, this review analyzes the new initiative by the World Health Organization (WHO) to assess fracture risk and new information on assessment of bone health in rheumatic disease patients. PMID- 17121494 TI - Update on treatment of arthritis in children: new treatments, new goals. AB - Although improved in recent years, the outcome for children who have juvenile idiopathic arthritis (JIA) is still less than ideal. Between 25% and 70% of children with JIA will still have active arthritis 10 years after disease onset, and over 40% will enter adulthood with active arthritis. Based on older publications, the fatality rate for JIA is 4- to 14-fold greater than an age- and sex-matched healthy population, and up to 39% of patients are significantly incapacitated, either wheelchair-bound or bedridden (Steinbrocker Classes III or IV). The hope is that recent changes in treatment approaches will result in marked improvement in long-term functional outcomes, although this has yet to be proven. JIA-associated chronic uveitis has a high frequency of serious complications: 20% develop cataracts, 19% glaucoma, and 16% band keratopathy. The anti-TNF biologics have all been tested in children with polyarticular JIA in blinded, placebo-controlled clinical trials, with over 70% in each trial demonstrating response that has been sustained in longer term follow-up studies. In systemic JIA approximately 50% respond to anti-TNF agents, but in many the response is not sustained. Openlabel studies have shown promising results for biologic therapies that block interleukins 1 and 6 in systemic JIA. PMID- 17121495 TI - Biomarkers in osteoarthritis. AB - Osteoarthritis (OA) can be a progressive disabling disease, which results from the pathological imbalance of degradative and reparative processes, with concomitant inflammatory changes. The synovium, bone, and cartilage are each well established sites involved in the pathophysiological mechanisms that lead to progressive joint degeneration. The search for disease-modifying osteoarthritis drugs, DMOADs, has been hampered by several factors, including the variable progression of disease, the lack of specificity and sensitivity of standard radiography, and the fact that the slowing of radiographic progression may not result in corresponding improvement in pain and function. As a result, there is general agreement that development of DMOADs will be facilitated by advances in imaging and the validation of chemical biomarkers. Such biomarkers should be useful tools that will identify patients at risk for disease progression and predict responses to candidate structure-modifying drugs. PMID- 17121496 TI - Gout in 2006: the perfect storm. AB - Mechanistic and therapeutic advances in gout have been moving swiftly in the past decade. Clinically, the disease is changing in character. This review discusses several of the pertinent recent developments in understanding gout and in novel therapeutics for the disease. Subjects addressed include the role of URAT1 mediated renal proximal tubule epithelial cell urate anion reabsorption in hyperuricemia. We discuss the therapeutic limitations of allopurinol and uricosurics and the potential applications of novel xanthine oxidase inhibitors and of recombinant uricase preparations. Last, we summarize understanding of the central role of the early induced innate immune response in gouty inflammation, which has suggested the potential value of new strategies for treating gouty inflammation by targeting caspase-1 or IL-1beta. PMID- 17121498 TI - Directed evolution and structural analysis of N-carbamoyl-D-amino acid amidohydrolase provide insights into recombinant protein solubility in Escherichia coli. AB - One of the greatest bottlenecks in producing recombinant proteins in Escherichia coli is that over-expressed target proteins are mostly present in an insoluble form without any biological activity. DCase (N-carbamoyl-D-amino acid amidohydrolase) is an important enzyme involved in semi-synthesis of beta-lactam antibiotics in industry. In the present study, in order to determine the amino acid sites responsible for solubility of DCase, error-prone PCR and DNA shuffling techniques were applied to randomly mutate its coding sequence, followed by an efficient screening based on structural complementation. Several mutants of DCase with reduced aggregation were isolated. Solubility tests of these and several other mutants generated by site-directed mutagenesis indicated that three amino acid residues of DCase (Ala18, Tyr30 and Lys34) are involved in its protein solubility. In silico structural modelling analyses suggest further that hydrophilicity and/or negative charge at these three residues may be responsible for the increased solubility of DCase proteins in E. coli. Based on this information, multiple engineering designated mutants were constructed by site directed mutagenesis, among them a triple mutant A18T/Y30N/K34E (named DCase-M3) could be overexpressed in E. coli and up to 80% of it was soluble. DCase-M3 was purified to homogeneity and a comparative analysis with wild-type DCase demonstrated that DCase-M3 enzyme was similar to the native DCase in terms of its kinetic and thermodynamic properties. The present study provides new insights into recombinant protein solubility in E. coli. PMID- 17121500 TI - Broken Hill experience: a comparison to the United States' area health education centers' national network role and responsibilities in rural health. PMID- 17121501 TI - Relationships between the health care sector and the media. PMID- 17121502 TI - Value adding through regional coordination of rural placements for all health disciplines: The Broken Hill experience. AB - OBJECTIVE: To describe the experience of the University of Sydney's Broken Hill Department of Rural Health (BHUDRH) delivering a regional program in far western NSW. DESIGN: Case Study Setting: The Australian Government's University Departments of Rural Health Program, established in 1997, has been responsible for setting up regionally coordinated rural placement programs for students from all health disciplines in each state and territory. RESULTS: Over nine years the BHUDRH has harnessed both academic and administrative resources to achieve greater efficiency in program delivery, increased support for students during their placement, and enhanced educational opportunities that draw on regional issues. The Broken Hill program accepts students from 22 Australian universities and places more students in the region, for longer periods than previously (271 students in 2005 compared with 140 students in 1998). CONCLUSION: Regional coordination, linked to investments in rural-based infrastructure and support systems can provide a sustainable platform from which to provide quality rural placements for students from all health disciplines, while at the same time increasing regional capacity to take students. PMID- 17121503 TI - Farm-related injury event, social consequences and injury reporting in the Land Lantbruk newspaper in Sweden: a retrospective study of farm-related injury reporting during 2000-2005. AB - OBJECTIVE: The aim of the present study was to detect how a leading news paper, the Land Lantbruk in Stockholm, Sweden, informs the public and specifically the rural sector in Sweden and the Scandinavian countries concerning injury events (farm-related injury event) and the use of injury prevention and control. Injury reporting in the Land Lantbruk has been studied from the point of injury prevention and control. A study of injury prevention and rural health and safety in Australia shows that the newspaper The Land that serves Australia's rural community should 'be an under-utilised vehicle for news and commentary on rural health and safety issues'. DESIGN: The study period was from January 2000 to February 2005. A total of 178 articles were reviewed and analysed. The articles were available on a newspaper database in the Land Lantbruk newspaper. Articles that addressed farm-related injury event and rural health and safety were chosen and organised into subgroups. RESULTS: Tractor and motor vehicle safety (35%) was most common among the injury reporting. Although the newspaper Land Lantbruk provided excellent coverage of the causes of these events, the reports tended to focus on circumstances and did not provide information on injury prevention or the advantages of also coverage of the social and psychosocial long-term consequences of accidents. CONCLUSION: In the prevention work of reducing farm related injuries in the rural sector in the Scandinavian countries and decreasing the human suffering represented by this health problem, rural politicians, insurance companies, rural authorities and also handicap organisations should listen more to the injured individuals and their own experiences relative to the difficulty of life after an accident. The reaction of family and relatives, and experiences of the long-term social consequences, have not been included in the media coverage. Journalists at the Land Lantbruk could also share experiences of the Swedish coverage of rural health and safety from Australian journalists from The Land. PMID- 17121504 TI - Total joint replacement surgery in a rural centre. AB - OBJECTIVE: To demonstrate that total joint replacement surgery can be safely and effectively performed in rural hospitals with acceptable outcomes. DESIGN: Case series. SETTING: A rural district hospital. PARTICIPANTS: PARTICIPANTS were 64 patients, 30 men and 34 women, who underwent total knee replacements (TKR); and 63 patients, 41 men and 22 women, who had total hip replacements (THR). MAIN OUTCOME MEASURES: Level of patient satisfaction following total joint replacement surgery, obtained by patient interview. Incidence of postoperative joint specific complications, for example infection, THR dislocation and manipulation under anaesthetic (MUA) of a TKR. RESULTS: None of the TKR or THR patients developed a deep wound infection. In this study 8.8% TKR patients had an MUA but all during a period of limited physiotherapy services; 5.8% THR patients suffered a dislocated prosthesis. Following TKR 95.3% patients reported to be 'happy' with the outcome of their surgery. Of the THR patients 97.0% declared they were 'happy' with their surgical outcome. CONCLUSIONS: There was a high level of patient satisfaction, low infection rate, acceptable levels of MUA for TKR and dislocation for THR following total joint replacement in our rural district hospital. The surgeons performed a medium volume of total joint replacements and an appropriate multidisciplinary team was in place. In such settings joint replacement surgery can be safely and successfully performed in rural centres to the benefit of rural patients, surgeons and GPs. PMID- 17121505 TI - Suturing as an advanced skill for registered nurses in the emergency department. AB - OBJECTIVE: Many patients present to emergency departments (EDs) for wounds that require repair by suturing. This study looked at ascertaining the efficacy of Advanced Clinical Nurses' (ACN) suturing dermal lacerations in tertiary, regional and rural ED settings. DESIGN: Convenience sample; prospective questionnaire. SETTING: Three mixed adult and paediatric EDs. These departments were tertiary, regional and rural regional EDs. PARTICIPANTS: Eighty patients requiring sutures for dermal lacerations that were repaired by an ACN. Of them 24 were men and seven were women. Mean age was 40.5 years, with a range of 21-58 years. Ten results from rural ED; six from regional ED; 15 from tertiary ED. MAIN OUTCOME MEASURE(S): Any differences between suturing standards in rural, regional and tertiary EDs; clinical results of suturing performed by ACNs; local medical officers' satisfaction with wound repair. RESULTS: A total of 31 responses were received. The lacerations were located on the face in 13 patients (42%), the scalp in 10 (32%), an extremity in 2 (6%), and on the trunk in 6 (20%). A total of 196 sutures were required. Local medical officers graded wound repair as 'good' in 24 cases (77%) and adequate in seven cases (23%). There were no wound healing complications reported. CONCLUSIONS: Nurses who complete a standardised training program in wound management and repair are capable of providing high quality, definitive care for patients who present to EDs with dermal lacerations. This is true irrespective of whether the Registered Nurse is working in a rural, regional or tertiary ED. PMID- 17121506 TI - Patient outcomes following an intervention involving community pharmacists in the management of depression. AB - OBJECTIVE: Documentation and evaluation of patient outcomes in a pilot study into the role of rural community pharmacists in the management of depression. DESIGN: Parallel groups design with a control and intervention group. SETTING: Thirty-two community pharmacies in rural and remote New South Wales, Australia. PARTICIPANTS: One hundred and six patient participants, mean age of 46 years, predominantly female, not currently employed, recruited by participating pharmacists. INTERVENTIONS: Intervention pharmacists were given video-conference training on the nature and management of depression by a psychiatrist, psychologist and general practitioner and asked to dispense medication with extra advice and support. Control pharmacists were asked to provide usual care. MAIN OUTCOME MEASURES: Adherence by self-report, K10, Drug Attitude Index. RESULTS: The results indicated that adherence to medications was high in both groups (95% versus 96%) and that both groups had improved significantly in wellbeing (a reduction K10 score of 4 (control) versus 4.7 (intervention)). No significant change was found in attitude to drug treatment once baseline scores were controlled for. CONCLUSIONS: Because both groups improved in wellbeing it is not possible to claim that the training provided to the intervention pharmacists was responsible for the success. However, the improvements gained in such a short time (two months) suggest that the involvement of pharmacists has had a beneficial rather than negative effect. Further research into the most appropriate ways in which to integrate the skills of pharmacists into a model of mental health care delivery in rural communities is recommended. PMID- 17121507 TI - Aeromedical evacuations from an east Arnhem Land community 2003-2005: the impact on a primary health care centre. AB - OBJECTIVE: To understand the profile and impact of aeromedical evacuations in remote Indigenous communities. DESIGN: Descriptive study. SETTING: A primary health care centre in east Arnhem Land, Northern Territory, Australia. PARTICIPANTS: Four hundred and ten evacuations from a total population of more than 2200 were analysed from February 2003 to August 2005. MAIN OUTCOME MEASURES: Patient demographics, seasonal variations, diagnostic categories, utilisation of staff resources. RESULTS: On average 6.5% of the community were evacuated to hospital every year with an evacuation occurring every 2.2 days. Children aged under five years were 3.3-fold overrepresented in evacuations (comprising 37.7% of those evacuated versus 11.3% of the community, P < 0.001). Four diagnostic categories accounted for 61% of evacuations: respiratory disease (21%), obstetric conditions (15%), gastroenteritis (14%) and injury/poisoning (11%). Over the study period four patients required intubation at the clinic. Evacuation rates were higher in the monsoon season. Forty-seven per cent of evacuations occurred after hours. The waiting time for plane arrival ranged from one hour to 21 hours with a median wait-time of three hours. CONCLUSION: Aeromedical evacuations place a heavy burden on primary health centres. Clinic staff are regularly required to provide hospital-level acute care, often for several hours at a time. Meeting this burden competes with primary prevention programs and regular clinic duties. The age and diagnostic profiles encountered in this study have significant implications for the range of skills required to provide an adequate acute care service. This study highlights the need for remote area health centres to be well resourced to meet these needs. PMID- 17121508 TI - Managing patients at risk of deterioration in rural hospitals: a qualitative study. AB - OBJECTIVE: This paper examines strategies used by nurses to manage patients at risk of deterioration in smaller rural hospitals in Victoria, Australia. DESIGN: A two-stage design comprising a postal questionnaire (n = 20) and case studies (n = 7). Case study participants were interviewed and their practice was observed for one shift. SETTING: Smaller rural hospitals in Victoria. PARTICIPANTS: Registered Nurses (Division 1). MAIN OUTCOME MEASURE: Observed and reported strategies used to manage deteriorating patients. RESULTS: Nurses in this study used clinical strategies and communication strategies when managing patients who might deteriorate. These strategies were adapted to suit the individual rural context. The ability to anticipate patient requirements and confidence to act on clinical presentation were central. Nurses also had to learn the art of referral and they tailored communication according to their perception of the knowledge and skills of medical colleagues. Strategies were also influenced by staffing patterns and expectations of the community and health service. CONCLUSIONS: We conclude that rural nurses play an essential role in assessing and managing risk of deterioration across a range of patient presentations and often in challenging situations. Strategies used by nurses required an understanding of the rural context of care delivery. PMID- 17121509 TI - A technically simple method for staining of acid-fast bacilli in cytology smears: an evaluation. AB - OBJECTIVE: To study the effects of modifications in the Ziehl-Neelsen staining procedure on predictive accuracy for acid fast bacilli in comparison to the conventional technique. Simplicity of procedure and reagent economy were the factors taken into consideration. DESIGN: Comparative evaluation between thick and thin air-dried smears stained conventionally and thick ethanol-fixed smears stained by the modified technique was done. RESULTS: Positive predictive accuracy of all the three smears, that is, thick air-dried, thin air-dried and thick ethanol-fixed, was 100%. Negative predictive accuracy for thick air-dried, thin air-dried and thick ethanol-fixed smears was 36.36%, 32.33% and 34.78%, respectively. Overall predictive accuracy was 66.67% for thick air-dried, 61.90% for thin air-dried and 64.29% for thick ethanol-fixed. These differences were found to be statistically insignificant. CONCLUSION: The modified method offers an accuracy comparable to the conventional technique, is simpler and with improved reagent economy. It is of special importance to diagnostic facilities in rural set-ups. PMID- 17121510 TI - Potentially pathogenic spotted fever group rickettsiae present in Western Australia. PMID- 17121511 TI - Under-use of bisphosphonates in rural elderly women who have sustained fractures. PMID- 17121512 TI - Community participation in rural health care, a quality perspective. PMID- 17121518 TI - Phaeochromocytoma, new genes and screening strategies. AB - Following recent advances in the genetics of phaeochromocytomas and paragangliomas, the members of the European Network for the Study of Adrenal Tumours (ENS@T) Phaeochromocytoma Working Group have decided to share their genotyping data and to propose European recommendations for phaeochromocytoma/functional paraganglioma (PH/FPGL) genetic testing. Germline DNA from 642 patients was analysed by ENS@T teams. In 166 patients (25.9%) the disease was familial and caused by germline mutations in VHL (56), SDHB (34), SDHD (31), RET (31) or NF1 (14), causing von Hippel-Lindau disease, SDHB- or SDHD PH/FPGL syndromes, multiple endocrine neoplasia type 2 (MEN 2) and type 1 neurofibromatosis (NF1), respectively. In almost 60% of inherited cases it was possible to formulate a probable genetic diagnosis based on family history and/or typical syndromic presentation. Genetic testing revealed mutations in 12.7% of cases with an apparently sporadic presentation. Several clinical characteristics, such as young age at onset, the presence of bilateral, extra-adrenal or multiple tumours or a malignant tumour, should be seen as indications for genetic testing. The ENS@T Phaeochromocytoma Working Group recommends the genetic testing of all patients with PH and FPGL and suggests a practice algorithm for the management of their exploration. PMID- 17121519 TI - Effect of thyroxine replacement on serum IGF-I, IGFBP-3 and the acid-labile subunit in patients with hypothyroidism and hypopituitarism. AB - OBJECTIVE: To describe the effect of T4 replacement in patients with primary and central hypothyroidism on components of the IGF binding protein complex: IGF-I, the acid-labile subunit (ALS) and IGFBP-3. PATIENTS AND METHODS: We determined IGF-I, ALS and IGFBP-3 (by 125I-IGF-II ligand blots and immunoblots) in serum of 19 patients with primary and 11 patients with central hypothyroidism. RESULTS: Mean (+/- SD) free T4 (fT4) increased from 4.4 +/- 2.4 pmol/l at baseline to 18.6 +/- 5.2 pmol/l following T4 therapy. In patients with primary hypothyroidism, IGF I concentrations increased from 101 +/- 57 to 158 +/- 60 microg/l (P < 0.001) and ALS from 12.6 +/- 4.7 to 15.6 +/- 5.2 mg/l (P = 0.001). IGFBP-3 levels (in arbitrary units, AU), assessed by 125I-IGF-II ligand blot and by Western blot (the intensity of the 45/42-kDa doublet following T4 replacement defined as 1 AU) increased from 0.74 +/- 0.47 to 1 (P = 0.029) and from 0.76 +/- 0.42 to 1 (P = 0.018), respectively. In patients with hypopituitarism, IGF-I and ALS concentrations increased on T4 therapy from 49 +/- 23 to 97 +/- 36 microg/l (P < 0.001) and from 7.8 +/- 4.1 to 11.0 +/- 2.7 mg/l (P = 0.010), respectively. IGFBP 3 remained unchanged during T4 replacement. CONCLUSIONS: T4 replacement increases the serum levels of IGF-I and ALS in patients with primary as well as central hypothyroidism. IGFBP-3 levels increase in response to T4 replacement in patients with primary hypothyroidism but not in those with central hypothyroidism, suggesting that thyroid hormones increase IGF-I and ALS but not IGFBP-3 in patients with GH deficiency. PMID- 17121520 TI - Percutaneous coronary intervention increases leptin and decreases adiponectin levels. AB - OBJECTIVE: The study was designed to examine the effect of percutaneous coronary intervention (PCI) on adiponectin and leptin levels. We have previously demonstrated that PCI triggers a systemic inflammatory response. We hypothesized that inflammation participates in the pathogenesis of diabetes mellitus and the metabolic syndrome by modulating levels of adiponectin and leptin. DESIGN: Prospective study in which inflammation was induced by PCI. PATIENTS: Forty-eight patients with stable coronary artery disease and without diabetes mellitus. MEASUREMENTS: High-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), leptin and adiponectin were measured at baseline and 48 h after the procedure. RESULTS: Following PCI, hs-CRP increased by 211%, IL-6 by 87% and leptin by 19%, while adiponectin decreased by 14% (P < 0.001 for all). The change in IL-6 correlated with that in hs-CRP (rho = 0.32; P = 0.027), as did the changes in IL 6 and leptin (rho = 0.31; P = 0.03). The change in adiponectin, however, did not correlate with the change in any of the other markers. CONCLUSION: This study demonstrates that PCI affects the levels of adiponectin and leptin within 48 h. These effects may be secondary to the inflammatory response triggered by PCI. PMID- 17121521 TI - Effect of recombinant hGH (rhGH) replacement on gonadal function in male patients with organic adult-onset GH deficiency. AB - OBJECTIVE: Previous evidence indicated that, in adults with organic hypopituitarism, GH deficiency (GHD) may mask the presence of other pituitary deficits, in particular central hypothyroidism and hypoadrenalism. Little and conflicting information is available about the relationship between GHD, rhGH therapy and gonadal function in males. The aim of the present study was to investigate the hypothalamic-pituitary-gonadal axis (HPG) in male adults with organic GHD and normal HPG axis. PATIENTS: Twelve male adults (mean age 48 +/- 7 years) with organic GHD and normal HPG axis. MEASUREMENTS: Serum levels of testosterone, LH and FSH (basal and after GnRH stimulation test), SHBG and IGF-I and percentage body fat (BF%) were evaluated before and during rhGH (mean dose 0.24 +/- 0.02 mg/day for 13 +/- 1 months) treatment. RESULTS: Serum IGF-I levels normalized during rhGH treatment and BF% significantly decreased. Serum testosterone levels significantly decreased (from 18.1 +/- 1.7 to 14.2 +/- 1.6 nmol/l, P = 0.01), with a parallel and significant decrease of serum SHBG (from 31.1 +/- 3.6 to 24.3 +/- 2.3 nmol/l, P < 0.05). Thus, calculated free testosterone (cFT) did not change (from 0.39 +/- 0.17 to 0.33 +/- 0.14 nmol/l, P = ns). Finally, no difference was found in basal and GnRH stimulated gonadotrophins levels. CONCLUSIONS: In conclusion, the condition of GHD does not seem to mask central hypogonadism, in contrast to what is observed for central hypothyroidism and hypoadrenalism. However, the significant decrease in serum testosterone levels, strictly related to SHBG decrease, suggests that evaluation of the HPG axis during rhGH treatment cannot be based on the measurement of total testosterone levels, but should mainly rely on calculation of cFT and a careful clinical evaluation, in order to avoid unnecessary replacement therapy. PMID- 17121522 TI - Effects of pioglitazone and metformin on plasma adiponectin in newly detected type 2 diabetes mellitus. AB - OBJECTIVE: This prospective study evaluates the effect of insulin sensitizers, pioglitazone (PGZ) and metformin (MET) on plasma adiponectin and leptin levels in subjects newly diagnosed with type 2 diabetes mellitus (T2DM). DESIGN: Double blind, randomized, active control, dose escalation study of 12 weeks treatment duration. PATIENTS: Thirty apparently healthy, treatment-naive T2DM patients diagnosed within the past 6 months. MEASUREMENTS: Plasma adiponectin and leptin levels were estimated by enzyme-linked immunosorbent assay (ELISA), and insulin resistance by the homeostasis model of assessment (HOMA-IR). RESULTS: Baseline plasma levels of adiponectin were lower in diabetic (n = 30) subjects than matched controls (n = 10, 6.6 +/- 1.1 vs 10.4 +/- 4.2 microg/ml, P = 0.021). The 12-week treatment with PGZ significantly increased adiponectin concentrations (6.6 +/- 1.1-17.9 +/- 7.4 microg/ml, P < 0.001) with no alteration in the MET treated group (6.8 +/- 1.5-6.7 +/- 2.8 microg/ml, P = 0.9). A significant decrease in plasma leptin levels was observed in the MET treated group (32.0 +/- 28.9-21.4 +/- 23.3 ng/ml, P = 0.024) but not in the PGZ treated group (23.9 +/- 24.1-22.4 +/- 25.4 ng/ml, P = 0.69). The alterations in plasma adiponectin and leptin levels were not associated with any change in body mass index (BMI). PGZ therapy improved insulin sensitivity to a greater degree (P = 0.007 and P = 0.001 for fasting plasma insulin (FPI) and HOMA-IR, respectively) than MET (P = 0.75 and P = 0.02 for FPI and HOMA-IR, respectively) but this improvement was not significantly different from that of MET at the end of 12 weeks (P = 0.146 and P = 0.09 for FPI and HOMA-IR, respectively). However, improvement in insulin sensitivity with PGZ was not commensurate with the increase in adiponectin. Better control of postbreakfast plasma glucose (PBPG) as well as decrease in serum triglycerides (TGs) were also seen with PGZ (PBPG, P < 0.001; TGs, P = 0.013). The rest of the parameters were comparable. Adverse reactions reported were minor and did not result in treatment discontinuation. CONCLUSIONS: Pioglitazone therapy appears to be better in achieving glycaemic control and increasing plasma adiponectin and insulin sensitivity in newly detected type 2 diabetics. PMID- 17121523 TI - Determinants of life expectancy in medullary thyroid cancer: age does not matter. AB - OBJECTIVE: In medullary thyroid cancer (MTC) age is considered an important prognostic factor but survival has never been properly adjusted for baseline mortality in the general population. We aimed to identify prognostic factors by analysing patients with MTC regarding life expectancy. DESIGN: We described a retrospective cohort study with a median follow-up of 8 years (range 1-35 years). PATIENTS: We included 120 consecutive patients of whom 66 (55%) had sporadic MTC. Male/female ratio was 1 : 1; median age was 45 years (range 3-83 years). MEASUREMENTS: Measurements were overall and disease-specific survival and life expectancy expressed as survival adjusted for baseline mortality rate in the general population. RESULTS: Overall and disease-specific 10-year survival was 65% and 73%, respectively. After 10 years, 29% of patients were biochemically and 63% clinically cured. Median overall life expectancy was 0.58 (95%CI 0.37-0.80). Detectable recurrence occurred in 60 patients after a median of 36 months (range 5-518 months). On multivariate regression analysis only stage of disease and extrathyroidal extension predicted recurrence-free life expectancy. Extrathyroidal extension was the only independent predictor of overall life expectancy. Persistent biochemical MTC did not independently affect life expectancy but calcitonin doubling time of less than one year indicated worse prognosis. Patients without detectable recurrences after initial treatment had a life expectancy similar to the general population. CONCLUSIONS: In MTC patients, extrathyroidal extension and stage at diagnosis are the only independent predictors of (recurrence-free) life expectancy. Patients diagnosed in an early stage of disease and patients without detectable recurrence have favourable life expectancy independently of biochemical cure. PMID- 17121524 TI - Glucose tolerance and lipid profile in longterm exogenous subclinical hyperthyroidism and the effects of restoration of euthyroidism, a randomised controlled trial. AB - OBJECTIVE: The impact of prolonged subclinical hyperthyroidism on glucose and lipid metabolism is unclear. Therefore, we evaluated glucose and lipid metabolism in patients with differentiated thyroid carcinoma (DTC) on TSH suppressive thyroxine therapy as a model for subclinical hyperthyroidism and investigated whether restoration to euthyroidism affects metabolism. DESIGN: We performed a prospective, single-blinded, placebo-controlled, randomised trial of 6 months duration with 2 parallel groups. PATIENTS: Twenty-five subjects with a history of differentiated thyroid carcinoma with > 10 years TSH-suppressive therapy with l thyroxine completed the study. l-thyroxine dose was replaced by study medication containing l-thyroxine or l-thyroxine plus placebo. Medication was titrated to establish continuation of TSH suppression (low-TSH group, 13 patients) and euthyroidism (euthyroidism group, 12 patients). MEASUREMENTS: We evaluated glucose metabolism by glucose tolerance test and HOMA (IR) and lipid metabolism by lipid profile. In addition, we measured plasma concentrations of glucoregulatory hormones. RESULTS: At baseline, glucose tolerance, HOMA (IR), lipid profile and plasma concentrations of glucoregulatory hormones were within the normal range. No significant differences between the low TSH and euthyroidism group were observed. After 6 months, neither glucose nor lipid metabolism in the low TSH group were different from baseline values. CONCLUSION: In summary, glucose and lipid metabolism in patients with DTC and long-term subclinical hyperthyroidism in general are not affected. Restoration of euthyroidism in general does not affect glucose and lipid metabolism. PMID- 17121525 TI - Alpha-enolase: a novel autoantigen in patients with premature ovarian failure. AB - OBJECTIVE: Although controversial, the presence of circulating antiovarian antibodies (AOA) may be considered a marker of autoimmune premature ovarian failure (POF). The purpose of the present work was to evaluate the presence of AOA in POF patients, and to identify a possible autoantigen in order to develop a reliable diagnostic tool that might help to determine the real prevalence of autoimmune POF. DESIGN: Non-randomised study. Blood sampling for determination of circulating AOA. PATIENTS: One hundred and ten patients with POF and 60 normally menstruating women with no record of autoimmune diseases (controls). MEASUREMENTS: Presence of circulating AOA was assessed by Western-blot, using cytosolic fraction from human ovarian homogenate as antigen. RESULTS: Twenty-one of 110 women with POF presented circulating antibodies directed toward an antigen of approximately 50 kD. Sixty control subjects proved negative. After purification and analysis by mass spectrometry, the antigen was identified as alpha-enolase. CONCLUSION: Determination of the presence of circulating antialpha enolase antibodies might be instrumental in identifying those patients who may present a putative defect in immunoregulation and therefore a possible autoimmune aetiolgy for POF. PMID- 17121526 TI - Plasma phospholipid transfer protein activity, a determinant of HDL kinetics in vivo. AB - OBJECTIVE: Phospholipid transfer protein (PLTP) is an important regulator in the transport of surface components of triglyceride-rich lipoprotein (TRL) to high density lipoprotein (HDL) during lipolysis and may therefore play an important role in regulating HDL transport. In this study we investigated the relationship of plasma PLTP activity with HDL metabolism in men. DESIGN AND METHODS: The kinetics of HDL LpA-I and LpA-I:A-II were measured using intravenous administration of [D3]-leucine, gas chromatography-mass spectrometry (GCMS) and a new multicompartmental model for HDL subpopulation kinetics (SAAM II) in 31 men with wide-ranging body mass index (BMI 18-46 kg/m2). Plasma PLTP activity was determined as the transfer of radiolabelled phosphatidylcholine from small unilamellar phosphatidylcholine vesicles to ultracentrifugally isolated HDL. RESULTS: PLTP activity was inversely associated with LpA-I concentration and production rate (PR) after adjusting for insulin resistance (P < 0.05). No significant associations were observed between plasma PLTP activity and LpA-I fractional catabolic rate (FCR). In multivariate analysis, including homeostasis model assessment score (HOMA), triglyceride, cholesteryl ester transfer protein (CETP) activity and PLTP activity, PLTP activity was the only significant determinant of LpA-I concentration and PR (P = 0.020 and P = 0.016, respectively). CONCLUSIONS: Plasma PLTP activity may be a significant, independent determinant of LpA-I kinetics in men, and may contribute to the maintenance of the plasma concentration of these lipoprotein particles in setting of hypercatabolism of HDL. PMID- 17121527 TI - Increased plasma concentrations of N-terminal pro-B-type natriuretic peptide in patients with mild primary hyperparathyroidism. AB - OBJECTIVE: Primary hyperparathyroidism (PHPT) is associated with heart disease. The aims of the present study were to evaluate how cardiac function and secretion of N-terminal pro-B-type natriuretic peptide (NT-proBNP) correlate in patients with mild PHPT, and how the plasma level of NT-proBNP is influenced by cure of the parathyroid disease. DESIGN AND PATIENTS: Forty-two patients with PHPT without symptoms of heart disease were examined before and 1 year after curative parathyroidectomy. MEASUREMENTS: Plasma or serum concentrations of NT-proBNP, calcium, PTH, creatinine, oestradiol, testosterone and SHBG were measured. Cardiac function was evaluated by equilibrium radionuclide angiography (ERNA). RESULTS: At baseline, NT-proBNP levels correlated negatively with systolic function [left ventricular ejection fraction (LVEF), P < 0.001]. Twelve per cent of the patients had NT-proBNP levels above normal reference values preoperatively. One year postoperatively, the corresponding proportion was 21%. The mean plasma concentration of NT-proBNP increased after parathyroidectomy (P < 0.01) in parallel with a dip in diastolic function (peak filling rate, P < 0.05) and a falling trend in systolic function (LVEF, P = 0.08). The postoperative percentage changes in circulating NT-proBNP and total oestradiol correlated positively (P < 0.05). CONCLUSIONS: Patients with mild PHPT and normal renal function may have high levels of circulating NT-proBNP despite the absence of symptomatic heart disease. Cure of the parathyroid disease is followed by a further increase in NT-proBNP secretion in parallel with ERNA measures, indicating subclinical changes in heart function. These results are in line with data indicating an association between PHPT and increased risk of premature death. PMID- 17121528 TI - Natural course of benign thyroid nodules in a moderately iodine-deficient area. AB - OBJECTIVE: Few studies using ultrasonographic evaluation have addressed the natural course of benign thyroid nodules. In this study, we present the long-term analysis of benign, untreated thyroid nodules in a moderately iodine-deficient area. DESIGN: We collected the retrospective data for 531 nodules in 420 patients followed by ultrasonography for a mean time period of 39.7 +/- 27.8 months (range 12-168 months). RESULTS: When we considered significant nodule size decrease criteria as decreases in nodule volume of at least 15% and 30%, we found that 33.1% and 20.7% of thyroid nodules decreased in size, respectively. Using the criteria of a greater than 15% and 30% increase in volume showed that 32% and 24.1% increased in size, respectively. A logistic regression analysis using mixed models to predict thyroid nodule growth (volume change > or = 15%), including age, TSH, sex and time between evaluations, failed to predict nodule growth. Only hypoechoic echo pattern remained as a statistically significant predictor of growth. CONCLUSION: In 40 months' follow-up, we found that benign thyroid nodules exhibited a mostly favourable clinical natural course. One-third of benign thyroid nodules showed continuous growth, one-third remained unchanged, and the other third decreased in size. PMID- 17121529 TI - Do serum leptin levels have a role in the prediction of pregnancy outcome in case of threatened miscarriage? AB - OBJECTIVE: To determine whether maternal serum leptin levels can predict pregnancy outcome in cases of threatened miscarriage. DESIGN: Observational study. PATIENTS: Fifty-seven women with normal pregnancy (group A), 19 women with threatened miscarriage who delivered at term (group B), 22 women with threatened miscarriage with subsequent miscarriage (group C) and 27 women with delayed miscarriage (group D). MEASUREMENTS: Maternal serum leptin was determined using a commercial radioimmunoassay (RIA) kit. RESULTS: Serum leptin levels were not significantly different among groups A, B and C [median (range) 17.6 (10.1-26.5), 18.2 (12.5-25.6) and 17.3 (11.2-27.6) microg/l, respectively]. Women in group D showed significantly lower serum leptin levels in comparison with other groups [14 (6.7-21.3) microg/l; P < 0.05]. CONCLUSIONS: Serum leptin levels in the first trimester of pregnancy may not be the primary indicator of miscarriage in cases of threatened abortion. PMID- 17121530 TI - Elevated circulating adiponectin in type 1 diabetes is associated with long diabetes duration. AB - OBJECTIVE: To study circulating adiponectin concentrations in relation to diabetes duration and endogenous insulin secretion in patients with type 1 diabetes. PATIENTS: Patients with haemoglobin A1c (HbA1c) < 6% (reference range 3.6-5.4%) were selected for the study. Twenty-two men and 24 women [age 41.3 +/- 13.8 years (mean +/- SD), diabetes duration 4 months to 52 years] participated. Healthy controls (15 women and nine men, age 41.3 +/- 13.0 years) were also included. Overnight fasting serum samples were analysed for adiponectin, HbA1c, C peptide and lipoproteins. RESULTS: Significant positive associations were found between adiponectin concentrations and diabetes duration in univariate and multiple regression analyses. Serum adiponectin averaged 9.7 +/- 5.3 [median 8.1, interquartile range (IQR) 3.6] mg/l in patients with diabetes duration less than 10 years and 17.8 +/- 10.7 (median 14.7, IQR 7.5) mg/l in patients with longer duration (P = 0.0001). Among the patients, 24 were without detectable (< 100 pmol/l) and 22 with detectable C-peptide levels (185 +/- 91 pmol/l). C-peptide levels in controls averaged 492 +/- 177 pmol/l. HbA1c was 5.7 +/- 0.6% in patients without detectable C-peptide and 5.6 +/- 0.4% in patients with detectable C-peptide (ns). Serum adiponectin was higher in patients without detectable C-peptide than in patients with detectable C-peptide [17.3 +/- 11.1 vs. 10.6 +/- 5.8 mg/l (P < 0.005)] and in the controls [10.1 +/- 2.9 mg/l (P < 0.001 vs. patients without detectable C-peptide)]. CONCLUSIONS: The increase in circulating adiponectin concentrations in patients with type 1 diabetes appears to be strongly associated with long diabetes duration, irrespective of the metabolic control. Among other factors, a putative role for residual beta-cell function in the regulation of circulating adiponectin levels can be considered but we did not find sufficient evidence for this in the present study. PMID- 17121531 TI - Bone mineral density in patients with growth hormone deficiency: does a gender difference exist? AB - OBJECTIVE: The aim of the study was to clarify whether a gender difference exists with respect to bone mineral density (BMD) and bone mineral content (BMC) in adult patients with growth hormone deficiency (GHD). DESIGN: A case-control design. METHODS: Blood sampling for measurements of calcium, phosphate, creatinine, PTH, vitamin D, IGF-1, markers of bone formation and bone resorption, and dual energy X-ray absorptiometry (DEXA), to determine BMD and BMC of the lumbar spine, hip, distal arm and total body, were performed in 34 patients with GHD (19 females) and 34 sex-, age- and weight-matched healthy control subjects. The patients were well substituted on all pituitary axes, apart from GH. RESULTS: GH-deficient males had significantly lower BMD in the lumbar spine (P = 0.02), hip (P = 0.01) and total body (P = 0.003) than healthy males while GH-deficient females compared to healthy females had identical BMD values at all regions. This gender difference was even more obvious when BMD values were expressed as Z scores or as three-dimensional BMD of the total body. The bone formation and bone resorption markers, as well as calcium and vitamin D, were all at the same levels in GH-deficient and healthy males, indicating identical bone turnover. The GH deficient females, however, had significantly lower levels of bone markers compared to healthy females, indicating a reduced bone turnover. Oestrogen substitution of the GH-deficient females could explain this difference. CONCLUSIONS: Compared to healthy control subjects GH-deficient males had, in contrast to GH-deficient females, significantly reduced BMD and BMC. This obvious gender difference seems to be caused by the oestrogen substitution given to the females, compensating for the lack of GH, an effect testosterone does not seem to possess. PMID- 17121532 TI - Expression of activin and inhibin subunits, receptors and binding proteins in human adrenocortical neoplasms. AB - OBJECTIVE: The growth and differentiation factors activin and inhibin can affect tumour formation and steroid production in the adrenal cortex. These factors bind to type I (Alk-4), type II (ActRIIA, ActRIIB) and type III (betaglycan) receptors or to the activin-binding protein follistatin. Expression of these activin related mRNAs was measured in different types of adrenocortical tissues and tumours to study the relationship with tumorigenesis. DESIGN: Quantitative expression of activin-related mRNAs was investigated in patient adrenocortical samples. PATIENTS: Twenty-eight human adrenocortical samples from normal and hyperplastic adrenals and from adrenocortical adenomas and carcinomas were collected after surgery for study purposes. MEASUREMENTS: Using quantitative reverse transcription polymerase chain reaction (RT-PCR), we investigated the expression of inhibin alpha-, betaA- and betaB-subunits, follistatin, betaglycan, ActRIIA, ActRIIB and Alk-4 in the adrenocortical tissues. The expression of cytochrome P450c17 (CYP17) mRNA was also measured to investigate its association with inhibin and activin subunit expression. RESULTS: All genes studied were expressed in all tissues, with the exception of the inhibin alpha-subunit in one hyperplastic adrenal and three adrenocortical carcinomas. Expression of inhibin betaA-subunit, follistatin, betaglycan, ActRIIA, ActRIIB and CYP17 differed between nontumorous adrenals and carcinomas. CONCLUSIONS: These differences, together with correlation analysis, indicate parallel regulation of the expression of CYP17, the inhibin alpha-subunit, ActRIIA, ActRIIB, betaglycan and follistatin. We conclude that the expression of activin and inhibin subunits, receptors and binding proteins is affected by tumour formation in the adrenal gland and may play a role in tumorigenesis. PMID- 17121533 TI - 11Beta hydroxysteroid dehydrogenase type 1 is expressed and is biologically active in human skeletal muscle. AB - OBJECTIVE: No data exist regarding the distribution and oxoreductase enzyme activity of 11beta hydroxysteroid dehydrogenase type 1 (11beta HSD-1) in fresh human skeletal muscle. We aimed to investigate the mRNA and protein expression of 11beta HSD-1 in fresh skeletal muscle, confirm its biological activity and determine its relationship with hexose-6-phosphate dehydrogenase (H6PDH). We also examined the muscle fibre localization of 11beta HSD-1. DESIGN: Eleven non diabetic community volunteers underwent muscle biopsy of vastus lateralis. MEASUREMENTS: (i) 11beta HSD-1 and H6PDH mRNA expression by quantitative reverse transcription polymerase chain reaction (RT-PCR); (ii) protein localization and fibre type specificity by immunohistochemistry; and (iii) enzyme oxoreductase activity by percentage conversion of 3H cortisone to cortisol. RESULTS: 11beta HSD-1 mRNA was expressed at low levels compared to human liver. Mean DeltaCT of skeletal muscle in 11 subjects was 19.57 (range 18.40-20.79) compared to DeltaCT of 12.75 in human liver, which equates to an approximate 100-fold higher level of expression. H6PDH mRNA was also detected with a mean DeltaCT of 14.46 (range 13.13-16.60), approximately 35-fold more abundant than 11beta HSD-1 in skeletal muscle. There was a significant correlation between 11beta HSD-1 and H6PDH (r = 0.67, P = 0.03). 11beta HSD-1 immunostaining was present in all muscle specimens, with similar distribution among fast and slow twitch fibres. 11beta HSD-1 oxoreductase activity was demonstrated, with mean conversion of cortisone to cortisol of 17.7% per 200 mg of muscle per 24 h (range 7.1-29.5%). CONCLUSIONS: 11beta HSD-1 mRNA and protein is expressed in fresh human skeletal muscle along with readily demonstrable oxoreductase activity. 11beta HSD-1 localization is not muscle fibre type specific. High levels of skeletal muscle H6PDH should ensure that oxoreductase activity predominates in vivo. PMID- 17121534 TI - Mutational analysis of the PTH 3'-untranslated region in parathyroid disorders. AB - OBJECTIVE: Sequence alterations in untranslated regions (UTRs) of genes are important contributors to human diseases, including hereditary thrombophilia, hereditary hyperferritinaemia-cataract and fragile X mental retardation syndromes. Recently, functional studies of the 3'-UTR of the PTH gene, encoding parathyroid hormone, have highlighted it as a potential target for pathogenic mutations in patients with parathyroid dysfunction. Regulation of PTH gene expression occurs in part through protein binding to a specific 26 nucleotide instability element in the 3'-UTR of PTH mRNA, in a sequence-dependent manner. Thus, the PTH 3'-UTR has emerged as an important potential contributor to parathyroid dysfunction. Therefore, we sought to rigorously examine the PTH 3' UTR in patients with primary and secondary parathyroid disorders, including primary parathyroid hyperplasia, secondary parathyroid hyperplasia, sporadic parathyroid adenoma and familial hypoparathyroidism of unknown genetic basis. PATIENTS AND DESIGN: Twenty-one parathyroid glands from 14 patients with primary parathyroid hyperplasia, 40 sporadic parathyroid adenomas from 40 patients, 42 parathyroid glands from 29 patients with secondary parathyroid hyperplasia and peripheral blood leucocytes from 24 affected members of eight kindreds with familial hypoparathyroidism of unknown genetic basis were examined for mutations in the 3'-UTR of the PTH gene. RESULTS: No alterations from the normal sequence were detected in any of the 127 samples examined. CONCLUSIONS: Based on the absence of identifiable DNA sequence alterations in these forms of parathyroid dysfunction, it is unlikely that mutation of the PTH 3'-UTR contributes frequently to their pathogenesis. PMID- 17121535 TI - Congenital hypothyroidism caused by new mutations in the thyroid oxidase 2 (THOX2) gene. AB - OBJECTIVE: Congenital primary hypothyroidism (CH) occurs in one of 4000 births and in 20% of the cases CH is due to a defect in thyroid hormonogenesis. Candidate genes were examined to determine the precise aetiology of suspected dyshormonogenesis in CH. DESIGN: The genes that code for thyroid peroxidase (TPO), pendrin (PDS), sodium iodide symporter (NIS) and thyroid oxidase 2 (THOX2) were sequenced directly from genomic DNA. PATIENTS: Two girls found to have CH in the neonatal screening programme and suspected of having thyroid dyshormonogenesis were investigated to identify their molecular defect. RESULTS: Patient A had a novel heterozygous 1 bp insertion in the THOX2 gene (ins602g). This insertion results in a frameshift that predicts a premature stop at codon 300. Analysis of cDNA, transcribed from lymphocyte RNA, showed that this mutation causes skipping of exon 5, resulting in a frameshift and a premature stop at codon 254. The euthyroid mother was also a heterozygous carrier of the mutation whereas the father was homozygous for the wild-type THOX2 gene. In patient B, compound heterozygous mutations (ins602g-->fsX300 and D506N) were identified. D506N was present in one allele of the clinically unaffected mother and in a brother, whereas the euthyroid father was heterozygous for ins602g. Sixty normal individuals did not harbour the mutations. Sequencing of the TPO, PDS and NIS genes revealed no mutations. CONCLUSIONS: The identified THOX2 mutations, which have not been described previously, are the probable causes of CH in the patients. Mutations in the THOX2 gene should be considered as the molecular cause of CH in young patients with thyroid dyshormonogenesis. PMID- 17121536 TI - Functional effects of genetic variants in the 11beta-hydroxylase (CYP11B1) gene. AB - OBJECTIVE: We previously described an association between the -344C/T 5' untranslated region (UTR) polymorphism in the CYP11B2 (aldosterone synthase) gene and hypertension with a raised aldosterone to renin ratio (ARR); the same genetic variant is also associated with impaired adrenal 11beta-hydroxylase efficiency. The -344 polymorphism does not seem to be functional, so is likely to be in linkage with variants in CYP11B1 that determine the associated variation in 11beta-hydroxylase efficiency. We therefore aimed to determine whether there is an association between CYP11B1 variants and hypertension and/or an altered ARR. DESIGN AND MEASUREMENTS: We screened 160 subjects divided into four groups, normotensive controls, unselected hypertensive subjects, and hypertensive subjects with either a high (> or = 750) or low ARR (< or = 200), for variants in the coding region of CYP11B1 by single-stranded conformation polymorphism (SSCP) and direct sequencing. The effects of these variants on enzyme function were assessed by conversion of 11-deoxycortisol to cortisol and 11-deoxycorticosterone (DOC) to corticosterone. RESULTS: Eight novel missense mutations were identified in the CYP11B1 gene that alter the encoded amino acids: R43Q, L83S, H125R, P135S, F139L, L158P, L186V and T196A. In each case they were heterozygous changes. However, no mutations were identified that could account for hypertension and/or a raised ARR. The variants L158P and L83S severely impaired enzyme function while R43Q, F139L, P135S and T196A enzymes resulted in product levels that were approximately 30-50% that of wild-type levels. The variant enzymes H125R and L186V resulted in substrate-specific alterations in enzyme function. H125R decreased conversion of 11-deoxycortisol to cortisol and L186V increased 11 deoxycortisol conversion. Neither had an effect on the conversion of DOC to corticosterone. CONCLUSION: No variants were identified in the coding region of CYP11B1 that could account for hypertension and/or a raised ARR. However, this in vitro study identifies the importance of these affected residues to enzyme function and will inform subsequent studies of structure-function relationships. PMID- 17121537 TI - Novel mutation of the calcium sensing receptor gene in familial hypocalciuric hypercalcaemia and neonatal severe hyperparathyroidism. PMID- 17121538 TI - The influences of hyperprolactinaemia and obesity on cardiovascular risk markers: effects of cabergoline therapy. PMID- 17121539 TI - Thyroid volume and intrathyroidal blood flow increase during pregnancy. PMID- 17121540 TI - No evidence for coding region mutations in the retinoblastoma-associated Kruppel associated box protein gene (RBaK) causing familial hyperaldosteronism type II. PMID- 17121541 TI - Characterization of the Drosophila myeloid leukemia factor. AB - In human, the myeloid leukemia factor 1 (hMLF1) has been shown to be involved in acute leukemia, and mlf related genes are present in many animals. Despite their extensive representation and their good conservation, very little is understood about their function. In Drosophila, dMLF physically interacts with both the transcription regulatory factor DREF and an antagonist of the Hedgehog pathway, Suppressor of Fused, whose over-expression in the fly suppresses the toxicity induced by polyglutamine. No connection between these data has, however, been established. Here, we show that dmlf is widely and dynamically expressed during fly development. We isolated and analyzed the first dmlf mutants: embryos lacking maternal dmlf product have a low viability with no specific defect, and dmlf(-)- adults display weak phenotypes. We monitored dMLF subcellular localization in the fly and cultured cells. We were able to show that, although generally nuclear, dMLF can also be cytoplasmic, depending on the developmental context. Furthermore, two differently spliced variants of dMLF display differential subcellular localization, allowing the identification of regions of dMLF potentially important for its localization. Finally, we demonstrate that dMLF can act developmentally and postdevelopmentally to suppress neurodegeneration and premature aging in a cerebellar ataxia model. PMID- 17121542 TI - Transient alterations in granule cell proliferation, apoptosis and migration in postnatal developing cerebellum of CRMP1-/- mice. AB - Collapsin response mediator proteins (CRMPs) consist of five homologous cytosolic proteins that participate in signal transduction involved in a variety of physiological events. CRMP1 is highly expressed during brain development; however, its functions remains unclear. To gain insight into its function, we generated CRMP1(-/-) mice with a knock-in LacZ gene. No gross anatomical changes or behavioral alterations were observed. Expression of CRMP1 was examined by the expression of the knocked-in LacZ gene, in situ hybridization with riboprobes and by imunohistochemistry. CRMP1 was found to be highly expressed in the developing the cerebellum, olfactory bulbs, hypothalamus and retina. In adults, expression level was high in the olfactory bulbs and hippocampus but very low in the retina and cerebellum and undetectable in hypothalamus. To study potential roles of CRMP1, we focused on cerebellum development. CRMP1(-/-) mice showed a decrease in the number of granule cells migrating out of explants of developing cerebellum, as did treatment of the explants from normal mice with anti-CRMP1 specific antibodies. CRMP1(-/-) mice showed a decrease in granule cell proliferation and apoptosis in external granule cell layers in vivo. Adult cerebellum of CRMP1(-/-) did not show any abnormalities. PMID- 17121543 TI - The substrate specificity of tRNA (m1G37) methyltransferase (TrmD) from Aquifex aeolicus. AB - Transfer RNA (m(1)G37) methyltransferase (TrmD) catalyzes methyl-transfer from S adenosyl-L-methionine to the N(1) atom of G37 in tRNA. In Escherichia coli cells, TrmD methylates tRNA species possessing a G36G37 sequence. It was previously believed that G36 was the positive determinant of TrmD recognition. In the current study, we demonstrate that TrmD from Aquifex aeolicus methylates tRNA transcripts possessing an A36G37 sequence as well as tRNA transcripts possessing a G36G37 sequence. In contrast, tRNA transcripts possessing pyrimidine36G37 were not methylated at all. These substrate specificities were confirmed by an in vitro kinetic assay using 16 tRNA transcripts. The modified nucleoside and the position in yeast tRNA(Phe) transcript were confirmed by LC/MS. Furthermore, nine truncated tRNA molecules were tested to clarify the additional recognition site. Unexpectedly, A. aeolicus TrmD protein efficiently methylated the micro helix corresponding to the anti-codon arm. Because the disruption of the anti-codon stem caused the complete loss of the methyl group acceptance activity, the anti codon stem is essential for the recognition. Moreover, the existence of the D-arm structure inhibited the activity. Recently, it was reported that E. coli TrmD methylates yeast tRNA(Phe) harboring a sequence A36G37. Thus, recognition of the purine36G37 sequence is probably common to eubacteria TrmD proteins. PMID- 17121544 TI - Fission yeast Tor2 links nitrogen signals to cell proliferation and acts downstream of the Rheb GTPase. AB - The target of rapamycin (Tor) plays a pivotal role in cell growth and metabolism. Yeast contains two related proteins, Tor1 and Tor2. In fission yeast, Tor1 is dispensable for normal growth but is involved in amino acid uptake and cell survival under various stress conditions. In contrast, Tor2 is essential for cell proliferation; however, its physiological function remains unknown. Here we characterize the roles of fission yeast Tor2 by creating temperature sensitive (tor2(ts)) mutants. Remarkably, we have found that tor2(ts) mimics nitrogen starvation responses, because the mutant displays a number of phenotypes that are normally induced only on nitrogen deprivation. These include G1 cell-cycle arrest with a small cell size, induction of autophagy and commitment to sexual differentiation. By contrast, tor1Deltator2(ts) double mutant cells show distinct phenotypes, as the cells cease division with normal cell size in the absence of G1 arrest. Tor2 physically interacts with the conserved Rhb1/GTPase. Intriguingly, over-expression of rhb1(+) or deletion of Rhb1-GAP-encoding tsc2(+) is capable of rescuing stress-sensitive phenotypes of the tor1 mutant, implying that Tor1 and Tor2 also share functions in cell survival under adverse environment. We propose that Tor1 and Tor2 are involved in both corroborative and independent roles in nutrient sensing and stress response pathways. PMID- 17121545 TI - Multiple phytohormones influence distinct parameters of the plant circadian clock. AB - Circadian systems coordinate endogenous events with external signals. In mammals, hormone-clock feedbacks are a well-known integration system. Here, we investigated phytohormone effects on plant-circadian rhythms via the promoter:luciferase system. We report that many hormones control specific features of the plant-circadian system, and do so in distinct ways. In particular, cytokinins delay circadian phase, auxins regulate circadian amplitude and clock precision, and brassinosteroid and abscisic acid modulate circadian periodicity. We confirmed the pharmacology in hormone synthesis and perception mutants, as rhythmic expression is predictably altered in an array of hormone related mutants. We genetically dissected one mechanism that integrates hormone signals into the clock, and showed that the hormone-activated ARABIDOPSIS RESPONSE REGULATOR 4 and the photoreceptor phytochrome B are elements in the input of the cytokinin signal to circadian phase. Furthermore, molecular expression targets of this signal were found. Collectively, we found that plants have multiple input/output feedbacks, implying that many hormones can function on the circadian system to adjust the clock to external signals to properly maintain the clock system. PMID- 17121546 TI - Identifying novel substrates for mouse Cdk5 kinase using the yeast Saccharomyces cerevisiae. AB - Among the mammalian Cdk family members, Cdk5, activated by the binding of p35, plays an important role in the control of neurogenesis, and its deregulation is thought to be one of the causes of neurodegenerative diseases. Overproduction of Cdk5 and p35 in yeast cells causes growth arrest, probably because of hyperphosphorylation of yeast proteins. We screened mouse brain cDNA that could recover the growth of yeast cells overproducing Cdk5 and p35, hoping that such cDNA encodes a substrate or inhibitor of Cdk5. Mouse brain cDNA library was introduced into a yeast strain in which Cdk5, p35 and mouse cDNA were over expressed under the control of the GAL promoter, and cDNA plasmids were isolated from the transformants that recovered growth on galactose medium. The analysis of those plasmids revealed that they harbored cDNA that encodes neuronal proteins including SCLIP and CRMP-1, and those with unknown function. We found that Cdk5 could phosphorylate SCLIP and CRMP-1 in vitro and the two proteins in cultured cells showed a mobility shift depending on Cdk5 activity and the presence of specific Ser or Thr residues, indicating that SCLIP and CRMP-1 are likely substrates for Cdk5 in vitro and in cultured cells. Further screening with these systems will enable us to identify more novel substrates and regulators of Cdk5/p35, which will lead to the exploration of Cdk5 function in diverse cellular systems. PMID- 17121547 TI - Female infertility in mice deficient in midkine and pleiotrophin, which form a distinct family of growth factors. AB - Midkine and pleiotrophin form a family of growth factors. Mice deficient in one of the genes show few abnormalities on reproduction and development. To understand their roles in these processes, we produced mice deficient in both genes; the double deficient mice were born in only one third the number expected by Mendelian segregation and 4 weeks after birth weighed about half as much as wild-type mice. Most of the female double deficient mice were infertile. In these mice, the numbers of mature follicles and of ova at ovulation were reduced compared to numbers in wild-type mice. Both midkine and pleiotrophin were expressed in the follicular epithelium and granulosa cells of the ovary. The expression of these factors in the uterus was dramatically altered during the estrous cycle. The diestrus and proestrus periods were long and the estrus period was short in the double deficient mice, indicating the role of the factors in the estrous cycle. Furthermore, vaginal abnormality was found in about half of the double deficient mice. These abnormalities in combination resulted in female infertility. Therefore, midkine and pleiotrophin, together with their signaling receptors, play important roles in the female reproductive system. PMID- 17121549 TI - Oxygen - elixir of life or Trojan horse? Part 2: oxygen and neonatal anesthesia. PMID- 17121550 TI - Pediatric cardiac surgery: an historical appreciation. AB - Many of the early, classic pediatric cardiac surgical operations were named after their originators. Some of these continue to be performed in the original form, many in modified form and some are obsolete. The development of many of these important early operations is reviewed and they are placed in the context of their times. PMID- 17121551 TI - Value of bispectral index monitor in differentiating between moderate and deep Ramsay Sedation Scores in children. AB - BACKGROUND: Pediatric patients who undergo diagnostic radiological imaging studies routinely require moderate or deep sedation to a Ramsay Sedation Score (RSS) of 4 or 5, respectively. The correlation between moderate and deep RSS and bispectral index (BIS) in children has never been validated. This study was designed to determine whether the BIS values correlate with RSS of children sedated for diagnostic imaging studies. METHODS: After Institutional Review Board approval, data were prospectively collected on all children receiving pentobarbital alone for computed tomography or magnetic resonance imaging. On arrival in the recovery room, any child assigned a Ramsay score of 4 or 5 was subsequently monitored with the BIS. Receiver-operating characteristic (ROC) analysis was used to evaluate the discriminative ability of BIS in differentiating depth of anesthesia. RESULTS: A total of 86 children over 1 year of age were enrolled. Children with Ramsay 4 (moderate) and Ramsay 5 (deep) sedation scores showed no significant difference in age or pentobarbital dose (mg.kg(-1)). BIS values followed a normal Gaussian-shaped distribution for both Ramsay scores. There was no significant difference in mean BIS values between the Ramsay groups (P = 0.64). There was a wide variation in BIS values (range of 31 90), which demonstrates that the BIS monitor does not correlate with the clinically assigned RSS depth of anesthesia in children. The ROC curve for BIS is equivalent to a line of nondiscrimination. CONCLUSION: BIS has limited ability to distinguish between moderate and deep Ramsay sedation levels in children who receive pentobarbital sedation for diagnostic imaging studies. PMID- 17121552 TI - Lidocaine pretreatment for the prevention of propofol-induced transient motor disturbances in children during anesthesia induction: a randomized controlled trial in children undergoing invasive hematologic procedures. AB - BACKGROUND: We examined the effect of lidocaine pretreatment before propofol administration on the incidence of transient motor disturbances and on propofol requirements for anesthesia induction in infants and children undergoing repeated painful diagnostic and therapeutic hematological procedures. METHODS: A series of 358 children subgrouped according to the presence of a peripheral-vein or central venous catheter were randomly assigned to receive an intravenous dose of 2% lidocaine (2.0 mg.kg(-1)) or an equivalent volume of saline, 1 min before propofol (1.5-3.5 mg.kg(-1)) injected for anesthesia induction. RESULTS: The incidence of spontaneous movements was significantly lower in patients pretreated with lidocaine than in those receiving placebo (2.5% vs 29%; P < 0.001, by chi square test), as was the propofol induction dose (1.6 +/- 0.2 mg.kg(-1) vs 2.2 +/ 0.3 mg.kg(-1); P < 0.001) and pain at the injection site in patients peripheral vein catheter (12% vs. 54%; P < 0.001). Lidocaine administration also improved children's acceptance as reported by parents on the Observational Scale of Behavioral Distress administered 2 h after the procedure (6.5 +/- 2.5 vs. 9.4 +/- 3.3; P < 0.001). Bouts of coughing developed significantly more frequently after lidocaine pretreatment than after placebo (62.5% vs. 17.5%; P < 0.001). CONCLUSIONS: Because lidocaine pretreatment before the induction of propofol based anesthesia decreases propofol-induced motor disturbances, lowers hypnotic requirements and reduces pain at the injection site, without inducing untoward events, thus improving children's and parental acceptance, it should become standard practice in infants and children undergoing repeated painful diagnostic and therapeutic hematological procedures. PMID- 17121553 TI - New formulae for predicting tracheal tube length. AB - BACKGROUND: The aim of this study was to determine the accuracy of standard techniques for estimating oral and nasal tracheal tube length in children and to devise more accurate predictive formulae that can be used at the bedside. METHODS: Data were collected from 255 children who required tracheal intubation whilst on the Pediatric Intensive Care Unit over a period of 1 year. Age, weight, the final length of the tracheal tube and the internal diameter were documented. Patients with a tracheostomy were excluded from the study. RESULTS: Using linear regression the following formulae best predicted final tracheal tube length. For children over 1 year of age: Insertion depth (cm) for orotracheal intubation = age/2 + 13 Insertion depth (cm) for nasotracheal intubation = age/2 + 15 For children below 1 year of age: Insertion depth of orotracheal tube (cm) = weight/2 + 8 Insertion depth of nasotracheal tube (cm) = weight/2 + 9 CONCLUSIONS: Current Advanced Paediatric Life Support guidelines underestimate the appropriate tracheal tube lengths for orotracheal intubation in children over 1 year of age. Similarly, the novel weight-based formulae for tracheal tube lengths in children below the age of 1 year proved more accurate than standard reference charts. We therefore recommend that these new formulae are prospectively evaluated. PMID- 17121554 TI - Can Bispectral Index aid laryngeal mask placement in children? AB - BACKGROUND: Laryngeal mask airway (LMA) placement requires an adequate depth of anesthesia to prevent patient movement and adverse airway responses. Patient movement, coughing, stridor or laryngospasm at LMA placement may result in transient hypoxia, injury to the patient and prolong time spent in the anesthetic room. The Bispectral Index Score (BIS) is a relatively new tool that has not yet established its place in routine clinical pediatric anesthesia practice. One potential use may be to predict an adequate depth of anesthesia for successful intraoperative interventions such as LMA placement in children. METHODS: A total of 116 children aged between 1 and 16 years due to have an LMA placed under general anesthesia were enrolled into this prospective, blinded observational study. Complication or success of LMA placement was then analyzed in relation to the BIS value at the time of placement. Other clinical variables were also considered. RESULTS: The difference in BIS values in children with complicated compared with successful LMA placement was small and BIS therefore appears not to be a reliable predictor for either. Inexperience of the anesthetist and a history of recent upper respiratory tract infection (URTI) may be better indicators for complications. CONCLUSIONS: Bispectral Index Score appears, from this study, not to be a useful tool to help predict complications of LMA placement or to help reduce airway complications during LMA placement. Other markers may be more useful predictors of adverse airway events. PMID- 17121555 TI - Prospective, randomized comparison of laryngeal tube and laryngeal mask airway in pediatric patients. AB - BACKGROUND: While reports of the use of laryngeal mask airway (LMA)-Classic in great patient numbers are available, data on the use of the laryngeal tube (LT) in this age group is limited. The two devices are compared in a prospective randomized trial to evaluate success rates and quality of airway seal. METHODS: Sixty children, aged 2-8 years, scheduled for elective surgical interventions were randomized to be ventilated with LMA or LT. Standardized anesthesia was induced with fentanyl and propofol. Number of insertion attempts, time until first tidal volume and intraoperative tidal volumes, and peak pressures were recorded. Airway leak pressure was measured with cuff pressure adjusted to 60 cmH(2)O. RESULTS: Demographic data were comparable, average age in the LMA/LT group was 5.2 +/- 1.9/5.3 +/- 1.9 years. Insertion was successful in 29 of 30 patients in the LMA group (second attempt 8) and in all patients in the LT group (second attempt 3). Time until first tidal volume for LMA/LT was 23.1 +/- 7.3/19.2 +/- 8.6 s (P < 0.05). Peak airway pressures for LMA and LT were 15.3 +/- 3.4 and 17.1 +/- 4.0 cmH(2)O (P < 0.05) with tidal volumes of 10.2 +/- 2.2 and 10.2 +/- 1.9 ml.kg(-1), airway leak pressure was 19.2 +/- 8.6 cmH(2)O for LMA and 26.3 +/- 7.3 cmH(2)O for LT (P < 0.001). CONCLUSION: Insertion success rate is high with both LMA and LT in the age group studied. The airway leak pressure, serving as an estimate to judge quality of airway seal, is higher with the LT. PMID- 17121556 TI - Continuous monitoring of central venous oxygen saturation in neonates and small infants: in vitro evaluation of two different oximetry catheters. AB - BACKGROUND: Accurate assessment and monitoring of the cardiocirculatory function are essential during major pediatric and pediatric cardiac surgery. Monitoring of the central venous oxygen saturation (ScvO(2)) may be a better indicator of tissue oxygenation and derangement of cellular oxygen utilization than the more commonly used vital parameters. Therefore, we compared oxygen saturation measurements with thin fiberoptic oximetry catheters and standard blood gas oximetry in an in vitro setting. METHODS: Two different size continuous fiberoptic oximetry catheters (2-4-F) were inserted in an extracorporeal circuit filled with human red blood cells in normal saline (haematocrit 30%, flow 600 ml.min(-1)). The results of fiberoptic oximetry were then compared with standard blood gas oximetry for a wide range of different oxygen saturations using linear regression. RESULTS: The oxygen saturations found ranged from 9% to 100%. The results of the two different fiberoptic oximetry catheters correlated significantly (r = 0.99, P < 0.0001) with standard blood gas oximetry. CONCLUSION: The results of fiberoptic oximetry are nearly identical with standard blood gas oximetry for a wide range of different oxygen saturations. Thin oximetry catheters can be inserted percutaneously even in neonates and small infants. The continuous monitoring of ScvO(2) may be beneficial, especially in patients who are in danger of developing low cardiac output or sudden cardiovascular collapse. PMID- 17121557 TI - Hemodynamic changes during acute elevation of intra-abdominal pressure in rabbits. AB - BACKGROUND: The intra-abdominal pressure (IAP) may be increased during pneumoperitoneum for minimally invasive surgery, after high tension repairs of congenital abdominal wall defects, major abdominal surgery, liver transplantation, abdominal trauma, peritonitis or ileus. The aim of this study was to investigate hemodynamic changes during elevation of IAP using an experimental setting, which mirrors anatomical and physiological conditions of neonates and small infants as closely as possible. METHODS: In five fasted, anesthetized, mechanically ventilated and multicatheterized New Zealand rabbits, the IAP was gradually increased by intra-abdominal infusion of normal saline (total volume 1000 ml). At baseline and after each infusion of 100 ml normal saline cardiac output (CO, transcardiopulmonary thermodilution), pressure in the superior (SVCP) and inferior vena cava (IVCP), mean arterial pressure (MAP), peak airway pressure (PAP) and IAP was recorded. RESULTS: During the study, IAP, SVCP and IVCP increased significantly. IVCP was significantly higher than SVCP from timepoint 200 ml to study end. After abdominal decompression IAP, SVCP and IVCP decreased to baseline levels. Changes in MAP were not significant. CO increased significantly from baseline to timepoint 200 ml (peak value), remained nearly constant until timepoint 800 ml and decreased thereafter until the abdominal infusion ceased. After abdominal decompression CO returned to baseline level. SVCP, IVCP and PAP correlated significantly with IAP (SVCP, r = 0.73; IVCP, r = 0.97; PAP, r = 0.94; P < 0.0001). CONCLUSIONS: The hemodynamic changes caused by increased IAP cannot be recognized by routine monitoring of arterial blood pressure and transcutaneous oxygen saturation. The increase in central venous pressure may be misinterpreted as an elevation of cardiac preload. One major effect of a prolonged increase in IAP is a decreased CO. PMID- 17121558 TI - Ascites in infants with severe sepsis - treatment with peritoneal drainage. AB - BACKGROUND: Ascites in neonates and infants is usually caused by cardiac failure and urinary or biliary tract obstruction. The objective of this study was to characterize our experience with ascites as a complication of sepsis. METHODS: We retrospectively collected and analyzed data of patients treated in the intensive care unit (ICU) of the university-based children's hospital, in whom ascites developed during nosocomial sepsis. Ten infants admitted to the ICU in the first 2 days of life developed sepsis on the mean 31.5 (+/-21.9) postnatal day. Gram negative bacteria were the causative organism in nine cases, and Staphylococcus hemolyticus in one. Because of sepsis, reintubation and mechanical ventilation were necessary. All patients received broad spectrum antibiotics (including meropenem and ciprofloxacin), blood transfusions, catecholamines and intravenous immunoglobulin preparations. Ascites was observed on the median 13.5 day of sepsis (range 3-36), and severely compromised gas exchange. Continuous peritoneal drainage was applied by means of an intravascular catheter placed in the right lower abdominal quadrant. RESULTS: The mean drained fluid volume was 44.7 (+/ 20.4) ml.kg(-1).day(-1), drainage was continued for a median of 5.5 (range 1-56) day, and enabled significant reduction of ventilator settings 24 h after its implementation. No severe complications related to drainage occurred; six of 10 babies survived. CONCLUSIONS: Ascites can develop in infants with sepsis and cause respiratory compromise. Continuous drainage of ascitic fluid by means of an intravenous catheter is relatively safe and can improve gas exchange. PMID- 17121559 TI - Airway management in two of newborns with Pierre Robin Sequence: the use of disposable vs multiple use LMA for fiberoptic intubation. AB - In this article, we discuss the use of LMAs as a conduit to intubate the trachea of two Pierre Robin Sequence infants. Multiple use LMAs will admit larger diameter tracheal tubes (TT) than their disposable counterparts. Increased friction with the surface of the TT makes passing even small diameter tubes through the lumen of the disposable LMA difficult. PMID- 17121560 TI - Transpulmonary thermodilution hemodynamic monitoring for pheochromocytoma surgery in a child with complex congenital heart disease. AB - Hemodynamic alterations related to pheochromocytoma surgery may lead to decompensation with an associated cardiac malformation. Moreover, they are incompletely described in children because of potential hazards of invasive monitoring. We report transpulmonary thermodilution hemodynamic monitoring during pheochromocytoma surgery in a child with a complex congenital heart malformation. Comprehensive analysis of hemodynamic events was obtained that guided therapeutic decisions. Cardiac function was preserved and no complication occurred. PMID- 17121561 TI - Pitfalls in pediatric tracheostomy: a case report. AB - Tracheostomy is required in approximately 22% of children with Goldenhar syndrome because of upper airway obstruction. We report a case of problematic tracheostomy placement in a 24-week-old ex-premature infant with Goldenhar syndrome. This infant had potentially life-threatening ventilatory insufficiency following tracheostomy placement related to tracheostomy tube size and position. These complications may have been avoided by a change in practice in methods of assessment of tube position. PMID- 17121562 TI - Prolonged use of dexmedetomidine in an infant with respiratory failure following living donor liver transplantation. AB - We used dexmedetomidine for more than 2 months in a mechanically ventilated infant without serious adverse effects. An infant with liver cirrhosis of unknown cause underwent living donor liver transplantation at the age of 9 months. Long term mechanical ventilation was required postoperatively, and midazolam with fentanyl had been used to sedate the patient. They required increase to 1.7 mg.kg(-1).h(-1) and 3.5 microg.kg(-1).h(-1), respectively, which were still inadequate. On postoperative day 29, dexmedetomidine was added. The rate of dexmedetomidine infusion was increased gradually to 1.4 microg.kg(-1).h(-1). It was discontinued temporarily to exclude drug-induced liver dysfunction. However, without dexmedetomidine, adequate sedation level was unattainable. Liver dysfunction was likely to be attributed to cytomegalovirus infection and after restarting dexmedetomidine, the respiratory condition improved. He was extubated 10 weeks after the operation. Dexmedetomidine was successfully tapered off over the following 2 weeks with no signs of withdrawal. Dexmedetomidine was a useful sedative for an infant who required mechanical ventilation for a prolonged period of time. PMID- 17121563 TI - Anesthetic management of an infant with Perlman syndrome. PMID- 17121564 TI - Anesthetic management of a 2-year-old male with propionic acidemia. PMID- 17121565 TI - Lumbar sympathetic blockade in a patient with cutis marmorata telangiectatica congenita. PMID- 17121566 TI - Anesthesia in a child with medium-chain Acyl-CoA dehydrogenase deficiency. PMID- 17121567 TI - Penile erection during remifentanil anesthesia in children. PMID- 17121568 TI - Propofol fear: 'mommy, I don't want the white stuff'. PMID- 17121580 TI - All talk and no action. PMID- 17121582 TI - CpG methylation patterns in the IFNgamma promoter in naive T cells: variations during Th1 and Th2 differentiation and between atopics and non-atopics. AB - Interferon-gamma (IFNgamma) gene expression is tightly regulated in early life, and exaggerated negative control of IFNgamma production in CD4(+) T cells has been associated with risk for subsequent development of atopy. Recent studies have demonstrated hypermethylation of CpG sites in the IFNgamma promoter in neonates, a mechanism which in mice leads to strong suppression of IFNgamma gene transcription. In the present study, the methylation status of six CpG sites in the proximal promoter of the human IFNgamma gene was determined by bisulphite sequencing. Cell populations studied were Th1 or Th2 polarized cell lines derived from neonatal and adult CD4(+)/CD45RA(+) T cells, CD4(+) and CD8(+) naive T cells from cord blood of children followed to outcome age 2 for assessment of atopy status, and CD4(+) and CD8(+) naive T cells from 6 yr old and adult atopics and controls. We demonstrate that in vitro differentiation of CD4(+) T cells down the Th1 pathway (but not the Th2 pathway) is accompanied by progressive demethylation of CpG sites in the IFNgamma promoter, which is most marked in neonatal cells. Atopy development by age 2 was not associated with variations in methylation patterns in cord blood T cells. However, IFNgamma promoter methylation was reduced in CD8(+) T cells from atopic children in the age range in which hyperproduction of IFNgamma as recently been identified as a common feature of the atopic phenotype. The findings demonstrate the potency of IFNgamma promoter methylation as a mechanism for control of human IFNgamma gene expression, particularly during early life. Differential regulation of IFNgamma promoter methylation in T cells may be an important contributory factor in atopy development in childhood, and this possibility warrants further detailed investigation. PMID- 17121583 TI - Increased mutations of CD72 transcript in B-lymphocytes from adolescent patients with systemic lupus erythematosus. AB - Recent studies have shown that B cells play a central role in the pathogenesis of systemic lupus erythematosus (SLE). Abnormal expression of molecules engaging in B-cell receptor (BCR) signaling and resultant hyperactivity of B cells has been reported in both mouse models of lupus and patients with SLE. CD72 on B cells is unique in that it regulates BCR signaling both positively and negatively. We analyzed the expression of CD72 protein and mRNA in peripheral blood B cells from adolescent patients with SLE. The expression level of CD72 on B cells of the patients was decreased compared with that on B cells of controls. Sequence analysis of CD72 mRNA showed significantly increased nucleotide mutations, including both nucleotide substitutions and deletions. Almost all (95.6%) of the CD72 transcripts from the patients had different nucleotide sequences from those of the wild type. About half (41.3%) of the mutations were point mutations located close to the sequence of the immunoreceptor tyrosine-based inhibitory motif (ITIM), which negatively regulates BCR signaling. These results indicate that increased nucleotide mutation of CD72 mRNA accounts for the decreased expression level of CD72 in B cells, and it might be related to hyperactivity of B cells in patients with SLE. PMID- 17121584 TI - Haplotypes of surfactant protein C are associated with common paediatric lung diseases. AB - Surfactant protein C is part of the surfactant complex lining up the alveoles and thereby inhibiting collapse of the airways. In addition it is involved in innate immune responses. Rare polymorphisms within surfactant protein C have been linked to sporadic paediatric lung diseases, like proteinosis or interstitial lung diseases. One study in the Finnish population described association of common polymorphisms with neonatal respiratory syndrome. Other common lung diseases have not yet been investigated for association with this gene. The aim of this study was to test surfactant protein C for association with bronchial asthma and with severe respiratory syncytial virus associated diseases in infancy. The two common amino acid variants Asn138Thr and Asn186Ser were genotyped on 322 children with asthma, 131 children with severe respiratory syncytial virus associated diseases and 270 controls. Statistical analyses of single polymorphisms made use of the Armitage's trend test; haplotypes were calculated with FAMHAP and FASTEHPLUS. Polymorphisms were in Hardy-Weinberg equilibrium and in tight linkage equilibrium in all populations. Single polymorphisms showed no association with the diseases, however, surfactant protein C haplotypes were associated with severe respiratory syncytial virus associated diseases (p = 0.013). Furthermore, an inverse haplotype distribution was found between children with asthma and respiratory syncytial virus infection (p = 0.00025). The results of our study might suggest opposing roles of surfactant Protein C in the genetic predisposition for respiratory syncytial virus associated diseases vs. asthma. The causal mechanism for this observation has still to be shown. PMID- 17121585 TI - Polymorphisms of the Bcl-2 family member bfl-1 in children with atopic dermatitis. AB - T lymphocytes accumulating in the skin of patients with atopic dermatitis (AD) have a prolonged survival and are key mediators of this inflammatory disease. The anti-apoptotic bfl-1 gene is unique in that it is the only member of the Bcl-2 family that is transcriptionally regulated by inflammatory cytokines and might therefore be important in promoting the survival of effector T cells in patients with AD. The aim of this study was to determine whether polymorphisms in the bfl 1 gene are associated with a predisposition to childhood AD. Four bfl-1 gene, single nucleotide polymorphisms (SNPs) were studied by ARMS-PCR in 105 Caucasian children with moderately severe AD and 110 non-atopic adult controls. In addition to the known polymorphisms of exon 1 (+141*A/G, +202*G/T, +303*A/G), we described a novel polymorphism in the promoter region of the gene (-1182*G/C). We found a significant difference in bfl-1 +141 genotype [OR (95% CI) 5.1 (1.0-25.2)], as well as bfl-1-1182:+141:+202:+303 G:A:G:A/G:A:G:A diplotype frequencies [3.5 (1.0 12.2)] in AD (p < 0.05). The study thus provides evidence for an association between bfl-1 polymorphisms and the genetic predisposition to AD. PMID- 17121586 TI - Influence of SNPs in cytokine-related genes on the severity of food allergy and atopic eczema in children. AB - Although many single nucleotide polymorphism (SNP) studies have reported an association of atopy, allergic diseases and total serum immunoglobulin E (IgE) levels, almost all of these studies sought risk factors for the onset of these allergic diseases. Furthermore, many studies have analyzed a single gene and hardly any have analyzed environmental factors. In these analyses, the results could be masked and the effects of other genes and environmental factors may be decreased. Here, we described the correlation between four genes [interleukin (IL)-4 (C-590T), IL-4 receptor (A1652G), FCER1B (G6842A) and STAT6 (G2964A)] in connection with IgE production; the role of IL-10 (C-627A) as a regulatory cytokine of allergy; and the severity of food allergy (FA) and atopic eczema (AE) in 220 Japanese allergic children. In addition to these SNPs, environmental factors, i.e., patient's attitude, indoor environment, and so on, were also investigated in this study. Our study was retrospective, and the correlation was analyzed by our defined clinical scores divided into three terms: worst symptoms, recent symptoms and general amelioration at the most recent examination during the disease course. Our results indicated that IL-10 AA, the genotype with lower IL-10 production, is associated with higher IgE levels in the serum (p < 0.0001, estimate; 0.912). Marginal liver abnormalities were observed in the subject group with both FA and AE (p < 0.1191, estimate; 0.1490). Our defined clinical scores enabled evaluation of various aspects of disease severity. Based on the scores, while no single SNP selected in this study determined severity, the combination of the SNP with laboratory data and environmental factors appeared to determine severity. PMID- 17121587 TI - Patterns of sensitization in infants and its relation to atopic dermatitis. AB - Longitudinal studies in infant populations using validated diagnostic criteria of atopic dermatitis and sensitization are rarely reported, and disease definitions, testing procedures, age of study population and evaluation of objective markers vary between countries and studies. The objectives of this prospective birth cohort study were to investigate: (i) the prevalence, the cumulative incidence and the pattern of transient and persistent sensitization to common food- and aeroallergens in unselected infants, (ii) the association between sensitization and the development of atopic dermatitis (AD) and (iii) the association between selected perinatal risk factors with respect to AD and post-natal sensitization. During a one-year period a cohort of 562 unselected newborns was established and followed up at the age of 3, 6, 9, 12 and 18 months of age. At all time points infants were examined clinically and by histamine release (HR), total- and specific immunoglobulin E (IgE) and skin prick test (SPT). Sensitization ever to > or =1 allergen at 18 months of age was 59%, 50% and 6% using HR, IgE and SPT, respectively. A transient sensitization to > or =1 allergen was found in 47%, 42% and 4% and a persistent sensitization in 17%, 10% and 3%, respectively. Sensitization to environmental allergens was frequently observed in infancy when testing with HR and IgE. Results of SPT gave much lower frequencies. Reactivity to foods was more frequent than to aeroallergens. The dominant pattern was low level transient sensitization. This is important to know when sensitization tests are used in the course of examination of infants with eczematous skin diseases. Specific definitions of sensitization like persistent reactivity, high-level sensitization and poly sensitization were clearly associated with AD. A maternal history of AD was a valuable tool in predicting AD in early infancy; a similar finding was less obvious with regard to post-natal sensitization. Cord-specific IgE and cord-HR positive reactivity did not prove better tools than cord-total IgE in predicting AD within the first 18 living months. PMID- 17121588 TI - Continuing food-avoidance diets after negative food challenges. AB - Negative food challenges for follow-up in patients previously diagnosed with food allergy should logically be followed by a normal diet. However, all patients do not reintroduce the food. The aims of the study were to define the proportion of negative food challenge not followed by a normal diet, and to identify possible reasons for not reintroducing the food. Patients with a negative food challenge were sent a questionnaire by mail. Items in the questionnaire included the symptoms at diagnosis, the duration of the diet, the fear of an accidental reaction during the avoidance diet and how it influenced the social life. Patients were also asked if the food was reintroduced after the negative food challenge, and if not, for which reasons. In 25.4% of the questionnaires (18/71) respondents reported that the food was not reintroduced. Patients with a previous diagnosis of peanut allergy tended to reintroduce the food less frequently than patients allergic to other foods. Girls were found to significantly less frequently reintroduce the food than boys. However, neither the severity of the initial reaction, the anxiety of an accidental reaction during the avoidance diet, nor a prolonged avoidance diet did influence the decision to reintroduce the food. Among other reasons listed, fears of persistence of allergies, with recurrent pruritus or non-specific skin rashes after eating the food, were reported in 12.7% of the total number of questionnaires. Patients who reintroduced the food reported that their social life generally improved. One quarter of previously allergic patients continue a food avoidance diet despite a negative challenge. We suggest reassessing food consumption in all patients after a negative food challenge, and in those still avoiding the specific food to consider a repeated challenge test. PMID- 17121589 TI - Sublingual desensitization in children with congenital malformations and latex allergy. AB - The frequency of latex allergy in children requiring multiple surgery ranges from 16.7% to 65%. The aim of this study was to investigate the safety and efficacy of latex desensitization in a group of 10 patients with a history of multiple surgical procedures and clinically manifested allergy to latex. We selected 10 children (female-male ratio = 5:5), aged 4-16 yr (mean +/- s.d.: 9 +/- 4), with a history of multiple surgical procedures, adverse reactions to latex and positive skin test to latex and/or specific immunoglobulin E (IgE). Latex allergy diagnosis was confirmed by specific provocation tests (cutaneous, sublingual, mucous, conjunctival tests). Rush (4-day) sublingual desensitization was performed with increasing doses of latex extract (ALK Abello) under patients' tongue until the highest dose of 500 microg of latex. A maintenance therapy (10 drops of undiluted solution three times a week) was recommended. During the 2-yr follow-up mean values of specific IgG4 and IgE, eosinophilic cationic protein and total IgE did not show significant variations. Patients did not manifest any adverse effect during the rush phase and only two patients manifested mild local symptoms during the maintenance therapy. All the challenges showed a reduction in terms of percentage of positivity and mean scores. All the patients showed a reduction of the mean individual score (p < 0.001). Furthermore patients who needed dental examination or surgery underwent such procedures without the occurrence of symptoms. Our preliminary results show sublingual desensitization to latex can be an important therapeutic tool in the management of young allergic patients requiring multiple operations. PMID- 17121590 TI - Short-term growth and adrenal function in children with asthma treated with inhaled beclomethasone dipropionate hydrofluoroalkane-134a. AB - Inhaled beclomethasone dipropionate (BDP) with the propellant hydrofluoroalkane 134a (HFA) has been designed to be equivalent in terms of safety to chlorofluorocarbon (CFC)-formulated metered dose inhalers (MDI). The aim was to assess whether BDP HFA MDI 100 microg twice daily was equivalent to BDP CFC MDI 100 microg twice daily in terms of effects on short-term lower leg growth rate (LLGR) and measures of hypothalamic-pituitary-adrenal (HPA) function. The study consisted of a randomized double-blind cross-over trial with three active, a run in and two wash-out periods each consisting of 2 wk. The place of study was a secondary referral outpatient clinic. The subjects involved were 14 boys and 10 girls with asthma, aged 7-12 yr. They were all administered BDP HFA 100 microg, BDP CFC 100 microg and 200 microg twice daily. The outcome measures included LLGR and 24-h urine-free cortisol (UFC) and total cortisol metabolites (TCM). Mean (SD) LLGR during run-in and BDP HFA 100 microg, BDP CFC 100 microg and 200 microg twice daily periods were 0.43 (0.23), 0.09 (0.29), 0.10 (0.45) and 0.08 (0.27) mm/wk. The one-sided 97.5% confidence interval for the difference in LLGR between BDP HFA 100 microg and BDP CFC 100 microg was 0.24, thus, below the predefined criterion of 0.20 mm/week. Inter-period comparisons of active treatments showed no differences between means of LLGR, UFC or TCM. Though non-inferiority between BDP HFA and CFC 100 microg twice daily in terms of effects on LLGR was not found, equivalence was suggested by comparisons of LLGR during run-in and active treatments and by HPA function measures. PMID- 17121591 TI - Asthma and allergy medication use and costs among pediatric primary care patients on asthma controller therapy. AB - As observational studies in children initiating GINA-Step 3 therapies are scarce, we evaluated outcomes and costs in a primary care cohort. Two-yr retrospective cohort study included French children (age: 6-14) continuously followed in BKL Thales database who received > or =2 consecutive prescriptions for GINA-Step 3 therapy (=addition of montelukast or other controllers ('other'), such as increasing inhaled-corticosteroid dose (hICS), adding long-acting beta agonist (LABA), or ICS + LABA). After matching on gender and propensity score, medication use [rescue (short-acting beta agonists), acute (antibiotics (AB), oral corticosteroids (OCS)), allergy (antihistamines, nasal steroids) and other respiratory] was estimated via mean number of prescriptions and mean cost (per child/per month), and cost trends. During 12-month follow-up, children adding montelukast (n = 71) vs. 'other' (n = 213) had similar asthma rescue/acute and allergy medication use. Subgroup with asthma and allergic rhinitis (A + AR) adding montelukast used less OCS and AB (p = 0.014). Two-yr cost trends suggest stable asthma/allergy medication use in montelukast group (0.83 euro) compared with increase in 'other' (5.39 euro), which was driven by nasal steroid use [0.32 euro ('other') vs. -0.04 euro (montelukast), p = 0.0013]. In subgroup with A + AR decline in asthma/allergy medication use in montelukast group (-0.47 euro) vs. increase in 'other' (11.05 euro), p = 0.015, was driven by differences in AB and OCS (p = 0.04) and nasal steroid use (p = 0.001). Concomitant asthma/allergy medication use was similar in children adding montelukast or 'other' controllers (hICS, LABA, ICS + LABA), while children with allergic rhinitis on montelukast used less AB. Concomitant medication costs after addition of montelukast remained stable, while 'other' group experienced increase, especially in children with concomitant allergic rhinitis. PMID- 17121592 TI - A new small volume holding chamber for asthmatic children: comparison with Babyhaler spacer. AB - When a new holding chamber for administrating inhaled medication is to be marketed, it needs to be compared with existing chambers with two questions in mind: is this chamber well accepted by patients and is there an in vitro equivalence? We compared the new small volume non-electrostatic valved holding chamber, usable with all pressurized metered-dose inhalers and equipped with a funny facemask, Vortex (Pari GmbH, Germany), to the most frequently prescribed holding chamber in France, Babyhaler (GlaxoSmithKline Laboratories). Preferences were studied for 75 families with a child no more than 4 yr old, using standard questionnaires. An in vitro study assessed the delivered dose and the particle size distribution of two HFA beclomethasone dipropionate pressurized metered dose inhalers (Becotide 250 microg per dose and Nexxair 100 microg per dose) by dose uniformity sample apparatus and cascade impactor according to the European Pharmacopoeia. Vortex was preferred by 95% of the families because of its small size, its duck facemask, and its robust appearance. Among children able to give their opinion, 86% preferred Vortex to Babyhaler. In vitro, both holding chambers reduced the delivered dose of beclomethasone dipropionate and increased the quantity of particles smaller than 5 microm in diameter with both medications. A higher proportion of fine particles was obtained with Nexxair than with Becotide (p < 0.05) and with Vortex than with Babyhaler (p < 0.05). As expected, throat deposition is dramatically reduced for both drugs with both holding chambers. The in vitro difference in the particle size distribution of beclomethasone dipropionate with both holding chambers probably has no clinical influence. PMID- 17121593 TI - Childhood allergic disorders in Samsun, Turkey: discrepancy between reported and diagnosed. AB - Schoolchildren (n = 1310) randomly selected from 32 schools in Samsun, Northern Turkey, were screened using the International Study of Asthma and Allergies in Childhood questionnaire. The prevalence of wheezing and current (last 12 months) wheezing were 21% and 14%, respectively: 2.3% of this group had received the diagnosis of asthma by a physician. Allergic skin rash was described in 17.3% and rhinitis in 44.7%, while 2.6% had been diagnosed with eczema and 10.5%, with allergic rhinitis. Respiratory symptoms were more common among 6-7-yr-old children compared with those aged 13-14 yr, and tended to be more prevalent in urban and coastal regions. The discrepancy between the rate of allergic symptoms and diagnosed allergic disorders may indicate a need for increased public and professional awareness and screening for allergic disorders in this area. PMID- 17121594 TI - Authors reply to beta-lactam allergy in children. PMID- 17121595 TI - Meningococcal biofilm formation: structure, development and phenotypes in a standardized continuous flow system. AB - We show that in a standardized in vitro flow system unencapsulated variants of genetically diverse lineages of Neisseria meningitidis formed biofilms, that could be maintained for more than 96 h. Biofilm cells were resistant to penicillin, but not to rifampin or ciprofloxacin. For some strains, microcolony formation within biofilms was observed. Microcolony formation in strain MC58 depended on a functional copy of the pilE gene encoding the pilus subunit pilin, and was associated with twitching of cells. Nevertheless, unpiliated pilE mutants formed biofilms showing that attachment and accumulation of cells did not depend on pilus expression. Mutation and complementation analysis revealed that the type IV pilus-associated protein PilX, which was recently shown to mediate interbacterial aggregation, indirectly supported microcolony formation by contributing to pilus expression. A large number of PilX alleles was identified among genetically diverse meningococcal strains. PilX alleles differed in their propensity to support autoaggregation of cells in suspension, but not in their ability to support microcolony formation within biofilms in the continuous flow system. PMID- 17121596 TI - H4 acetylation does not replace H3 acetylation in chromatin remodelling and transcription activation of Adr1-dependent genes. AB - Histone acetylation regulates gene expression. Whether this is caused by a general increase in nucleosome fluidity due to charge neutralization or by a more specific code is still matter of debate. By using a set of glucose-repressed Adr1 dependent genes of Saccharomyces cerevisiae, whose transcription was previously shown to require both Gcn5 and Esa1, we asked how changes of histone acetylation patterns at the promoter nucleosomes regulate chromatin remodelling and activation. When the signal of glucose reduction reaches the cells, H4 acetylation is kept constant while an increase of H3 acetylation occurs, in an Adr1- and Gcn5-dependent manner. In cells lacking Gcn5 activity, the H3 acetylation increase does not occur and an unexpected increase of histone H4 acetylation is observed. Nevertheless, chromatin remodelling and transcription activation are impaired, suggesting that acetylation of H3 and H4 histones plays different roles. PMID- 17121597 TI - Quantitative trait loci analysis of egg and meat production traits in a red junglefowlxWhite Leghorn cross. AB - Egg and production traits are of considerable economic importance in chickens. Using a White Leghorn x red junglefowl F(2) intercross, standard production measures of liver weight and colour, egg size, eggshell thickness, egg taste and meat quality were taken. A total of 160 markers covering 29 autosomes and the Z chromosome were genotyped on 175-243 individuals, depending on the trait under consideration. A total of nine significant quantitative trait loci (QTL) and three suggestive QTL were found on chicken chromosomes 1, 2, 4, 5, 7, 8, 10, 12, E47W24 and E22C19W28. PMID- 17121598 TI - Postnatal changes in the expression of genes located in the callipyge region in sheep skeletal muscle. AB - The expression of five genes surrounding the callipyge (CLPG) mutation was analysed in skeletal muscles from lambs at one prenatal and two postnatal ages that coincide with the onset and establishment of muscle hypertrophy. Genotype specific changes in transcript abundance were detected for paternal allele specific DLK1 and PEG11 (the official symbol of the latter is RTL1) and the maternal allele-specific MEG3, PEG11AS and MEG8 when the mutation was inherited in cis. There were differences in the temporal and muscle-specific effects on expression between the maternal allele-specific genes and paternal allele specific genes. Maternal inheritance of the CLPG allele had a significant effect on the expression of MEG3 and MEG8 at prenatal and postnatal ages, whereas paternal inheritance of DLK1 and PEG11 only affected postnatal expression. Genotype-specific changes in PEG11AS expression were detected only in prenatal muscle. Maternal inheritance of the mutation caused similar changes in MEG3 and MEG8 expression in the semimembranosus, which undergoes hypertrophy, and the supraspinatus, which does not hypertrophy. Paternal inheritance of the mutation caused changes in PEG11 expression in both muscles, although the magnitude of expression in semimembranosus was more than 100-fold greater than in supraspinatus. DLK1 expression was upregulated in callipyge animals at both postnatal ages in the semimembranosus, but there was no effect of genotype on DLK1 expression in the supraspinatus at any age. Increased DLK1 expression was likely the primary cause of muscle hypertrophy, but a contribution of PEG11 to the phenotype cannot be ruled out based on gene expression. PMID- 17121599 TI - Results of a whole-genome quantitative trait locus scan for growth, carcass composition and meat quality in a porcine four-way cross. AB - A whole-genome quantitative trait locus (QTL) scan for 31 phenotypes related to growth, carcass composition and meat quality was conducted using 1187 progeny of a commercial four-way cross. Animals were genotyped for 198 microsatellite markers that spanned the entire porcine genome. QTL analysis was conducted to extract information from paternal and maternal meioses separately using a rank based nonparametric approach for half-sib designs. Nine QTL exceeded genome-wide significance: one QTL affecting growth (average daily gain on SSC1), two QTL influencing carcass composition (fatness on SSC3 and muscle mass on SSC15) and six QTL influencing meat quality (tenderness on SSC4 and SSC14; colour on SSC5, SSC6 and SSCX; and conductivity on SSC16). All but one of these coincided with previously reported QTL. In addition, we present evidence for 78 suggestive QTL with a combined false discovery rate of 40%. PMID- 17121600 TI - Association analysis of adiponectin and somatostatin polymorphisms on BTA1 with growth and carcass traits in Angus cattle. AB - This study tested positional candidate genes adiponectin (ADIPOQ) and somatostatin (SST) for effects on carcass traits in a commercially relevant cattle population. Both genes are located within a region of BTA1 previously reported to harbour quantitative trait loci (QTL) that affect marbling, quality grade, yield grade, ribeye area and weaning weight in Bos taurus x Bos indicus crosses. Except for the first intron of ADIPOQ, both genes, including over 2 kb upstream of the promoters, were sequenced in five registered Angus sires to identify polymorphisms. A variable copy duplication and three single nucleotide polymorphisms (SNPs) in ADIPOQ and one SNP in SST were genotyped and tested for association with 19 traits in a 14-generation pedigree of 1697 registered Angus artificial insemination sires representing all the major USA lineages of the breed. Linear models that parameterized predicted genetic merits in terms of allele substitution effects were fit by weighted least squares, and goodness-of fit tests were employed to differentiate causal mutations or polymorphisms in strong linkage disequilibrium (LD) with causal mutations from markers in weak LD with QTL. We confirmed the presence of QTL affecting marbling, ribeye muscle area and fat thickness in the vicinity of SST and ADIPOQ on BTA1 in Angus; excluded SST as underlying the ribeye muscle area QTL; and excluded ADIPOQ as underlying the marbling score QTL. However, association analysis provides very limited information about QTL location and has little intrinsic value when performed in the absence of linkage or LD analysis using flanking marker data to localize the QTL effect relative to positional candidate genes. PMID- 17121601 TI - A microsatellite linkage map of the blacklip abalone, Haliotis rubra. AB - There is considerable scope for genetic improvement of cultured blacklip abalone Haliotis rubra in Australia using molecular marker-assisted, selective-breeding practices. Such improvement is dependent on the availability of primary genetic resources, such as a genetic linkage map. This study presents a first-generation linkage map of H. rubra, containing 122 microsatellite markers typed in a single full-sib family. These loci mapped to 17 and 20 linkage groups for the male and female respectively, and when aligned, the consensus map represented 18 linkage groups. The male linkage map contained 102 markers (one unlinked) covering 621 cM with an average intermarker spacing of 7.3 cM, and the female map contained 98 markers (eight unlinked) covering 766 cM with an average intermarker spacing of 9.8 cM. Analysis of markers informative in both parents showed a significantly higher recombination rate in the female parent, with an average male-to-female recombination ratio of 1:1.45 between linked pairs of markers. This linkage map represents a significant advancement in the genetic resource available for H. rubra and provides a framework for future quantitative trait loci mapping and eventual implementation of marker-assisted selection. PMID- 17121602 TI - Saving feral horse populations: does it really matter? A case study of wild horses from Donana National Park in southern Spain. AB - In the 1980s, a conservation programme involving a feral horse population, the Retuertas horses from the Guadalquivir marshes, was started in the Donana National Park. The analysis of an extensive genetic survey of this population, which now numbers 100 animals, and 10 additional European and North African breeds using DNA polymorphisms from 22 microsatellites is presented. Highly significant fixation indexes were obtained for all pairwise comparisons between the Retuertas population and other breeds. A population neighbour-joining breed phenogram was built using different distance measures, but the Retuertas population failed to cluster with either of the two major clades of European and North African breeds, highlighting its uniqueness. In fact, the Retuertas population was positioned at the base of the trees, which were rooted using donkey samples. Furthermore, assignment tests and the individual Q-matrices obtained with the structure programme isolated the Retuertas breed from the other breeds with only four K groups. Interestingly, some local semi-feral horses, known as Marismeno, also currently living in the Guadalquivir marshes, have some microsatellite genotypes that fall well within the Retuertas cluster. This raises the possibility of incorporating horses from the Marismeno population in a future conservation programme. PMID- 17121603 TI - Origin and phylogeographical structure of Chinese cattle. AB - Complete mitochondrial D-loop sequences of 231 samples were used to explore the origin and genetic diversity of Chinese cattle. Phylogenetical analysis of these sequences revealed both Bos taurus and Bos indicus mitochondrial types in Chinese cattle. Four of the previously identified mitochondrial DNA lineages (T1-T4) were identified in the Bos taurus type, including lineage T1, which was found for the first time in Chinese cattle. Two lineages (I1 and I2) were identified in the Bos indicus type. Our results support the suggestion that the Yunnan-Guizhou Plateau is the domestication site of Chinese zebu. We also found evidence that Tibetan cattle originated from taurine and zebu cattle. The distribution pattern of Chinese cattle breeds was closely related to the geographical and climatic background. It was possible to divide Chinese cattle in this study into two major groups: northern and southern cattle. PMID- 17121604 TI - Polymorphisms within the coding region of the bovine luteinizing hormone receptor gene and their association with fertility traits. AB - Mutations within a number of genes have been associated with variations in fertility in various mammals. However, to date there have been no such associations reported for cattle. Herein, we describe three single nucleotide polymorphisms (SNPs) in the luteinizing hormone/choriogonadotropin receptor gene of cattle (Bos taurus). These polymorphisms include two missense mutations and one sense mutation, and all are located in areas of conserved synteny. When assessed in terms of haplotypes, these SNPs were significantly associated with variations in cattle fertility and production traits, most notably on calving interval, days to first service and production index (the UK economic index of milk yield measured in poundGB). PMID- 17121605 TI - Physiological and genetic characteristics of black-boned sheep (Ovis aries). AB - Here we report for the first time the discovery of sheep that have black bones and black muscles. The spectral pattern of pigment extracted from tissues of these black-boned sheep is similar to that of black-boned Chinese silky fowl. Additionally, black-boned sheep have significantly higher plasma colour, tyrosinase activity and kidney function than normal sheep. Synonymous nucleotide substitutions in the tyrosinase (TYR) and melanocortin 1 receptor (MC1R) genes were detected in black-boned sheep when compared with the corresponding sequences in normal sheep. In addition, a missense mutation (215T>C) in exon 2 of tyrosinase-related protein 1 (TYRP1) was detected in black-boned sheep, and this resulted in a putative valine-to-alanine substitution at codon 68 (Val68Ala). PMID- 17121606 TI - Characterization of the porcine FABP5 gene and its association with the FAT1 QTL in an Iberian by Landrace cross. AB - We have characterized and mapped the porcine fatty acid binding protein 5, epidermal (FABP5) gene. According to linkage and RH mapping, this gene is located close to the FABP4 (fatty acid binding protein 4, adipocyte) gene on swine chromosome 4. We resequenced 4.7 kb of the FABP5 gene in the parental population of an Iberian x Landrace cross (IBMAP), identifying seven SNPs arranged in two distinct FABP5 haplotypes. QTL and association analyses in the IBMAP population showed that this gene is strongly associated with fat deposition. QTL and haplotype analysis revealed that both FABP4 and FABP5 (clustered in mammals) are major candidate genes for the FAT1 QTL; the most likely position for the FAT1 QTL is between these two genes. Finally, our results suggest the presence of more than one QTL affecting fatness traits on porcine chromosome 4. PMID- 17121607 TI - A 2.5-Mb contig constructed from Angus, Longhorn and horned Hereford DNA spanning the polled interval on bovine chromosome 1. AB - The polled locus has been mapped by genetic linkage analysis to the proximal region of bovine chromosome 1. As an intermediate step in our efforts to identify the polled locus and the underlying causative mutation for the polled phenotype, we have constructed a BAC-based physical map of the interval containing the polled locus. Clones containing genes and markers in the critical interval were isolated from the TAMBT (constructed from Angus and Longhorn genomic DNA) and CHORI-240 (constructed from horned Hereford genomic DNA) BAC libraries and ordered based on fingerprinting and the presence or absence of 80 STS markers. A single contig spanning 2.5 Mb was assembled. Comparison of the physical order of STSs to the corresponding region of human chromosome 21 revealed the same order of genes within the polled critical interval. This contig of overlapping BAC clones from horned and polled breeds is a useful resource for SNP discovery and characterization of positional candidate genes. PMID- 17121608 TI - Comparative mapping reveals multiple rearrangements between pig chromosome 6 and human 19q13. PMID- 17121609 TI - Isolation and radiation hybrid mapping of 213 porcine microsatellites. PMID- 17121610 TI - Mapping of Toll-like receptor genes in rainbow trout. PMID- 17121611 TI - Mapping of genes in a region associated with upper temperature tolerance in rainbow trout. PMID- 17121612 TI - Construction and characterization of a duck bacterial artificial chromosome library. PMID- 17121613 TI - Mapping of nine porcine interferon regulatory factor genes. PMID- 17121614 TI - Evaluation of kallikrein 7 as a disease-causing gene for canine atopic dermatitis using microsatellite-based association mapping. PMID- 17121615 TI - Linkage and RH mapping of 10 genes to a QTL region for fatness and muscling traits on pig chromosome X. PMID- 17121616 TI - Identification of new MHC-DRB3 alleles from Indian (Bos indicus) cattle. PMID- 17121617 TI - Association of MITF with white spotting in Beagle crosses and Newfoundland dogs. PMID- 17121619 TI - Role of anti-tumour necrosis factor-alpha therapeutic agents in the emergence of infections. AB - There is increasing interest concerning the possible impact of anti-tumour necrosis factor (TNF)-alpha therapeutic agents on the emergence of infections. However, these agents do not seem to increase the incidence of adverse infectious events significantly. Published observations concern mostly infections of the urinary and upper respiratory tracts that develop in the setting of co morbidities, such as anterior or concomitant immunosuppressive treatment. Infliximab appears to increase the risk of tuberculosis, but this effect has not been observed with other anti-TNF-alpha agents. To better characterise the adverse infectious effects associated with these agents, physicians should be encouraged to notify the microbiological data relating to all cases. PMID- 17121620 TI - Control of endemic methicillin-resistant Staphylococcus aureus--recent advances and future challenges. AB - Although the exact burden of disease caused by methicillin-resistant Staphylococcus aureus (MRSA) remains largely unknown, most experts agree that MRSA infections are an important clinical and public health problem. Thousands of reports have been published during the last four decades concerning epidemiological and microbiological aspects of MRSA, but uncertainty remains about the best approach to prevent and control this worldwide plague, especially endemic MRSA. Epidemiological methods, e.g., risk scores for targeted screening upon admission, rapid molecular tests and pre-emptive isolation of high-risk patients, new decontamination regimens and restriction of certain antibiotic classes, are all promising approaches that may decrease MRSA cross-transmission; however, further evidence is needed before these strategies can be implemented on a wide scale. Control of community MRSA is an additional challenge for the future, requiring improved surveillance and contact tracing, as well as education and treatment of both infected cases and colonised contacts. This review summarises recent advances and studies that address these issues. Overall, it seems that there is no level of MRSA prevalence for which active control measures are no longer warranted. PMID- 17121621 TI - Direct detection of methicillin-resistant Staphylococcus aureus in clinical specimens by a nucleic acid-based hybridisation assay. AB - The occurrence of methicillin-resistant Staphylococcus aureus (MRSA) is still increasing worldwide and is associated with significant morbidity, mortality and hospital costs. Screening for MRSA plays a key role in limiting further nosocomial spread of this organism. Control measures require a rapid and sensitive test for direct detection of MRSA carriage. This study evaluated an easy-to-use PCR-hybridisation assay for the direct detection of MRSA in clinical swab specimens. In total, 508 pairs of swabs from 242 patients at risk for MRSA carriage were analysed by the standard culture method and the PCR assay. One swab was used for PCR and culture, while the second was used for culture only. Of the 508 pairs tested, 37 were positive by culture and 35 were positive by PCR. Among the 471 culture-negative specimens, 465 were negative by PCR and six were PCR positive. The PCR assay had a sensitivity of 94.59%, a specificity of 98.73%, a positive predictive value of 85.37%, and a negative predictive value of 99.57%. The PCR-hybridisation assay enabled reliable detection of MRSA carriage in c. 4 h, thereby allowing its effective use in an MRSA control strategy. PMID- 17121622 TI - Evaluation of three chromogenic media (MRSA-ID, MRSA-Select and CHROMagar MRSA) and ORSAB for surveillance cultures of methicillin-resistant Staphylococcus aureus. AB - Screening specimens were homogenised in saline 0.9% w/v before either direct inoculation or following enrichment in broth on three chromogenic media (MRSA-ID, CHROMagar MRSA and MRSA Select) and ORSAB medium for the detection of methicillin resistant Staphylococcus aureus (MRSA). In total, 102 of 466 specimens yielded MRSA on at least one medium. After incubation for 16-18 h, the sensitivity was 51%, 59%, 47% and 65% on MRSA-ID, CHROMagar MRSA, ORSAB and MRSA Select, respectively, compared with 82%, 75%, 67% and 80%, respectively, after 42 h, and 93%, 95%, 79% and not tested, respectively, following broth enrichment. There were significantly more MRSA colonies on MRSA-Select after 16-18 h than on ORSAB or MRSA ID (p 0.001 and 0.0022, respectively), whereas there were more MRSA colonies after 42 h on MRSA-ID and MRSA-Select than on ORSAB (p 0.0004 and 0.012, respectively). The specificity of the media for identifying MRSA based on the colour of colonies after incubation for 16-18 h was 100%, 99%, 99% and 100%, respectively, compared with 98%, 97%, 98% and 98%, respectively, after 42 h, and 100%, 99%, 100% and not tested, respectively, following broth enrichment. The speed of detection (mean time to report a positive result) was 1.65, 1.72, 2.31 and 1.35 days, respectively. For each of the three media tested following enrichment, the use of an enrichment broth increased the detection rate of MRSA by 16-24%. PMID- 17121623 TI - Staphylococcal chromosome cassette evolution in Staphylococcus aureus inferred from ccr gene complex sequence typing analysis. AB - Staphylococcal chromosome cassette mec (SCCmec) elements within major lineages of healthcare- and community-associated methicillin-resistant Staphylococcus aureus (MRSA) clones were characterised using intra-SCCmec multilocus sequencing. A strong correlation was observed between sequence- and PCR-based typing methods (p <0.001). However, phylogenetic analysis of the SCCmec locus using concatenated sequences evidenced few recombination events. Sequence type (ST)-SCCmec1 was found in SCCmec elements types I and IV, suggesting the evolution of an SCCmecI element into an SCCmecIV element. This coincided with the spread of the clone harbouring this SCCmec element into the community. No correlation was observed between ST-SCCmec lineage and MRSA lineage, confirming multiple acquisitions of SCCmec by S. aureus. This was exemplified by the SCCmecIV ST-SCCmec10 element, which was detected in all of the clonal complexes examined, including healthcare- and community-associated MRSA. The acquisition of this SCCmec element was five- to ten-fold more common than that of others. Models of MRSA clone evolution suggest that this SCCmec was first found in the paediatric clone. PMID- 17121624 TI - Antimicrobial susceptibility testing in European hospitals: report from the ARPAC study. AB - This observational study describes the antimicrobial susceptibility testing (AST) methods and interpretive criteria used in European hospitals during 2001, focusing specifically on detection of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). Of 263 hospitals that took part in the ARPAC study, 192 submitted data on AST. Of these, 89% (n = 170) routinely used a disk-diffusion AST method, 43% (n = 82) used a semi-automated method, and 70% (n = 135) routinely determined MICs. Hospitals in southern Europe were less likely to use disk-diffusion, but were more likely to use a semi automated method (p <0.001). In total, 173 (90%) interpreted AST results using CLSI breakpoints; 30% of these detected MRSA using unmodified CLSI disk-diffusion methods, while 35% used the unmodified CLSI agar-screening method for MRSA; 41% and 30% adhered to unmodified CLSI methodology for disk-diffusion and agar screening, respectively, to detect VRE. Some of the modifications made may have greatly reduced the ability of the tests to detect MRSA/VRE. For example, 20% of respondents used excessively high incubation temperatures and 13% used inadequate incubation times to detect MRSA by disk-diffusion, and 28% used Mueller-Hinton agar instead of brain-heart infusion agar in VRE screening plates. The majority of respondents stated that they followed CLSI guidelines, but a high proportion had modified the CLSI methods for detecting MRSA and VRE, which may compromise clinical management and antimicrobial resistance surveillance. PMID- 17121625 TI - Genetic and phenotypic differences among Enterococcus faecalis clones from intestinal colonisation and invasive disease. AB - This study investigated the differences among Enterococcus faecalis isolates from the intestinal compartment of healthy volunteers (n = 36), intensive care unit (ICU) patients (n = 29) and blood isolates (n = 31) from the same institution, in comparison with seven epidemic clones from other institutions. In general, isolates from colonised ICU patients and from bacteraemic patients showed higher rates of antimicrobial resistance than isolates from colonised healthy volunteers, particularly for erythromycin and aminoglycosides. The proportion of isolates/clone was 1.05 in the community, 2.63 in the ICU, and 1.47 among bacteraemic cases, suggesting low clonal variation in ICUs. Two clones, RENC1 and RENC2, were frequently found as intestinal colonisers of ICU patients, and RENC1 was also found to colonise healthy volunteers. These two clones were a cause of bacteraemia in the institution studied, and RENC2 was also detected in various other Spanish hospitals. Both RENC1 and RENC2 were esp+, bacteriocin producers, and were resistant to all antibiotics tested except vancomycin and ampicillin. RENC1 produced haemolysin whereas RENC2 produced protease. The ace, agg, cylA, esp and gelE genes were more common among colonising strains from ICU patients than among isolates from individuals in the community. In both colonised groups (ICUs and the community), 40-50% of isolates harbouring the gelE and cylA genes did not express the corresponding phenotypes. Thus, the study indicated that particular E. faecalis clones might be well-adapted to hospital environments, and that surveillance should be directed specifically towards rapid detection of these disseminating clones in order to prevent infections and clonal spread. PMID- 17121626 TI - Virulence genotype and nematode-killing properties of extra-intestinal Escherichia coli producing CTX-M beta-lactamases. AB - This study evaluated the virulence potential of Escherichia coli isolates producing CTX-M beta-lactamases. During a 24-month period, 33 extended-spectrum beta-lactamase (ESBL)-producing E. coli, including 14 CTX-M-producers, were isolated from urinary tract infections at Nimes University Hospital, France. The prevalence of 14 major virulence factors (VFs) was investigated by PCR and compared with the prevalence in a group of 99 susceptible E. coli isolates. Ten VFs were less prevalent (p <0.05) in the ESBL isolates than the susceptible E. coli, while iutA and traT were more prevalent in ESBL isolates (p <0.05). Moreover, the CTX-M-producing isolates had significantly fewer VFs than TEM producing isolates. A novel infection model using the nematode Caenorhabditis elegans was developed to assess the virulence properties of extra-intestinal pathogenic E. coli (ExPEC) strains in vivo. C. elegans infection assays, using 14 ESBL-producing E. coli and ten susceptible E. coli isolates, indicated that the ability to kill nematodes correlated with the presence of VFs, and that CTX-M producing isolates had relatively low virulence in vivo. Overall, the results suggested that hospital-acquired CTX-M-producing E. coli, although adapted for survival in an antibiotic-rich environment such as the hospital milieu, have a relatively low intrinsic virulence potential. PMID- 17121627 TI - The role of procalcitonin in a decision tree for prediction of bloodstream infection in febrile patients. AB - Bloodstream infection (BSI) in febrile patients is associated with high mortality. Clinical and laboratory variables, such as procalcitonin (PCT), may predict BSI and help decision-making concerning empirical treatment. This study compared two models for prediction of BSI, and evaluated the role of PCT vs. clinical variables, collected daily in 300 consecutive febrile inpatients, for 48 h after onset of fever. Multiple logistic regression (MLR) and classification and regression tree (CART) models were compared for discriminatory power and diagnostic performance. BSI was present in 17% of cases. MLR identified the presence of intravascular devices, nadir albumin and thrombocyte counts, and peak temperature, respiratory rate and leukocyte counts, but not PCT, as independent predictors of BSI. In contrast, a peak PCT level of >2.45 ng/mL was the principal discriminator in the decision tree based on CART. The latter was more accurate (94%) than the model based on MLR (72%; p <0.01). Hence, the presence of BSI in febrile patients is predicted more accurately and by different variables, e.g., PCT, in CART analysis, as compared with MLR models. This underlines the value of PCT plus CART analysis in the diagnosis of a febrile patient. PMID- 17121628 TI - Public health implications of using various case definitions in The Netherlands during the worldwide SARS outbreak. AB - This study analysed the consequences of deviation from the WHO case definition for the assessment of patients with suspected severe acute respiratory syndrome (SARS) in The Netherlands during 2003. Between 17 March and 7 July 2003, as a result of dilemmas in balancing sensitivity and specificity, five different case definitions were used. The patients referred for SARS assessment were analysed from a public health perspective. None of the patients referred had SARS, based on serological and virological criteria. Nevertheless, all 72 patients required thorough assessment and, depending on the results of the assessment, institution of appropriate prevention and control measures. Changing case definitions caused confusion in classifying cases. A centralised assessment of the reported cases by a team with clinical and public health expertise (epidemiological and geographical risk assessment) is a practical solution for addressing differences in applying case definitions. The burden of managing non-cases is an important issue when allocating public health resources, and should be taken into account during the preparation phase, rather than during an outbreak. This applies not only to SARS, but also to other public health threats, such as pandemic influenza or a bioterrorist episode. PMID- 17121629 TI - Human metapneumovirus infection among children in Taiwan: a comparison of clinical manifestations with other virus-associated respiratory tract infections. AB - This study compared the clinical, laboratory and radiological features of infections caused by human metapneumovirus (hMPV) with other respiratory viruses. Nasopharyngeal aspirates and throat swabs were obtained from children during a 9 week period. hMPV was the virus isolated most frequently, followed by adenovirus, influenza virus A, respiratory syncytial virus and influenza virus B. hMPV infected children were younger, and were more likely to be female, to present with feeding difficulties, a rash, tachycardia and a longer duration of fever, and to cough less frequently. Increasing interstitial infiltrates and hyperinflation were the most common radiological findings. None of the children required mechanical ventilation. PMID- 17121630 TI - Antimicrobial prophylaxis to prevent surgical site infections in a rural sub Saharan hospital. AB - A prospective cohort study was performed to collect baseline data concerning surgical site infections (SSIs) and antimicrobial prophylaxis (AMP) in a remote sub-Saharan district hospital. The SSI rate of 22% was high. Most (88%) of the patients received prophylaxis after incision, and only 5% within the 30-min period before incision. Of all pathogens isolated from SSIs, 60% were resistant to the agent administered. The antibiotics given most frequently were chloramphenicol (60%), aminopenicillins (23%) and benzylpenicillin (15%). Staphylococcus aureus (36%), Escherichia coli (5%) and enterococci (4%) were the pathogens isolated most commonly from SSIs. PMID- 17121631 TI - Effectiveness and nephrotoxicity of colistin monotherapy vs. colistin-meropenem combination therapy for multidrug-resistant Gram-negative bacterial infections. AB - A retrospective cohort study evaluated the effectiveness and nephrotoxicity of intravenous colistin monotherapy vs. colistin-meropenem combination therapy for patients with multidrug-resistant Gram-negative bacterial infections. Fourteen patients received intravenous colistin monotherapy and 57 received colistin meropenem. No significant differences were found concerning clinical response of the infection (12/14 (85.7%) vs. 39/57 (68.4%), p 0.32) and development of nephrotoxicity (0/14 (0%) vs. 4/57 (7%), p 0.58). A favourable association was revealed between survival and treatment with colistin monotherapy compared to colistin-meropenem (0/14 (0%) vs. 21/57 (36.8%) deaths, p 0.007), even after adjusting for the variables for which significant differences were found. PMID- 17121632 TI - Clonality of slime-producing methicillin-resistant coagulase-negative staphylococci disseminated in the neonatal intensive care unit of a university hospital. AB - Methicillin-resistant coagulase-negative staphylococci (MR-CNS) (n = 132), isolated from pre-term neonates, were analysed to determine their antibiotic resistance patterns, clonal distribution, biofilm production and the presence of the ica operon. All MR-CNS were multiresistant, and 89% produced slime. A major clone was identified (77 isolates) among 115 Staphylococcus epidermidis isolates. Ten of 16 Staphylococcus haemolyticus isolates also belonged to a single clone. Most (80%) slime-positive isolates possessed all the ica genes tested, while the remaining 23 (20%) had a variety of gene combinations. The entire ica cluster was detected in three of 15 slime-negative isolates. One major and two minor slime positive, multiresistant MR-CNS clones had disseminated among hospitalised pre term neonates. PMID- 17121633 TI - Specific and sensitive diagnosis of syphilis using a real-time PCR for Treponema pallidum. AB - A real-time PCR assay with a Taqman probe was developed that targeted the polA gene of Treponema pallidum. The test was validated using an analytical panel (n = 140) and a clinical panel of genital samples (n = 112) from patients attending a sexually transmitted infections clinic. High sensitivities and specificities of 94-100% were achieved using two real-time PCR platforms, the Rotor-Gene and the iCycler. The assay can be completed within 2 h, enabling reporting in <8 h. This fast and robust assay is suitable for implementation in routine laboratories for diagnosing primary syphilis. PMID- 17121634 TI - Pyrosequencing for rapid identification of carbapenem-hydrolysing OXA-type beta lactamases in Acinetobacter baumannii. AB - Carbapenem-resistant Acinetobacter baumannii isolates producing carbapenem hydrolysing oxacillinases are emerging worldwide. These enzymes are divided into four phylogenetic subgroups: OXA-23-like, OXA-51-like, OXA-24-like and OXA-58 like. A PCR-based approach linked to pyrosequencing analysis was developed to identify the genes for these beta-lactamases. Carbapenem-hydrolysing oxacillinases were rapidly and unambiguously identified in a collection of carbapenem-resistant clinical isolates of A. baumannii and Acinetobacter junii. Pyrosequencing may provide a rapid tool for identification of OXA variants, thus avoiding delays inherent in classical sequencing methods. PMID- 17121635 TI - Escherichia coli pacemaker-related endocarditis. PMID- 17121636 TI - What is the place of teicoplanin and linezolid in the treatment of prosthetic joint infections? PMID- 17121637 TI - EUCAST Technical Note on linezolid. PMID- 17121638 TI - The new emergence of exercise neurobiology. PMID- 17121639 TI - Effects of a physical exercise intervention on subjective physical well-being, psychosocial functioning and general well-being among office workers: a cluster randomized-controlled cross-over design. AB - The purpose of the study was to examine the effects of a workplace physical exercise intervention on subjective physical well-being, psychosocial functioning and general well-being. The study was a cluster randomized-controlled trial with the department (n=4) as the unit of randomization. The subjects (n=90) were office workers [mean age 45.7 (SD 8.5) years]. Psychosocial functioning and well being variables were measured by descriptive visual rating scales. The cross-over design consisted of one 15-week intervention period of light resistance training and guidance and another 15-week period of no training and no guidance. The statistical analysis was based on linear mixed models. The active component of the intervention, light resistance training, resulted in a slight, but statistically significant, increase in subjective physical well-being (P=0.015). At the average training time of 5 min/working day (25 min/week) the average increase during the 15-week period was 4 units (95% confidence interval (CI) 1-7) and 5% (95% CI 1-9). The physical exercise intervention had no effect on somatic symptoms, anxiety, self-confidence, mood, mental stress at work, working atmosphere, life satisfaction or meaning of life. Daily light resistance training, conducted during the working day, had a positive direction on subjective physical well-being among office workers. PMID- 17121640 TI - Motivation for physical activity of psychiatric patients when physical activity was offered as part of treatment. AB - This study examined motivation variables, self-determination and self-schema, in relation to physical activity, among psychiatric patients with experience with physical activity as part of their treatment. Participants were patients (N=109) from 15 psychiatric hospitals or day-care institutions. Data were collected by questionnaires. A positive relationship between physical activity level, positive experiences of the activity and higher degree of self-determination and exercise self-schema was expected. Intrinsically regulated motives (motivated by the experience of the activity in itself) were positively and significantly related to physical activity level and the experience of decrease in symptoms during physical activity, and extrinsically regulated motives were negatively correlated with physical activity level. Intrinsically regulated motives gave an odds ratio of 20.0 for being physically active rather than inactive. Holding an exercise self-schema gave an odds ratio of 6.1 for being physically active. The majority of the patients (57.4%) reported that physical activity decreased their illness symptoms, but a few (11.9%) reported negative effects. The findings demonstrated that psychiatric patients do not differ from the normal population in relation to motivational mechanisms, even if they may experience more barriers to physical activities because of their illness. Therefore, in trying to motivate psychiatric patients, it is important to make physical activity as intrinsically motivating as possible by focusing on the positive experiences of the activity itself, as well as helping to develop an exercise self-schema. PMID- 17121641 TI - The relation between motor activity and [3H]uridine uptake in the mouse brain. AB - Using microautoradiography ex vivo we tested the effect of forced running on a roller drum for 3 h on the nuclear incorporation of [5-(3)H uridine] in mouse brain. Specific neuron types with increased nuclear labelling included primary motor cortex layer 5 nerve cells with nuclei greater than 12 microm (+38%) and large neuron nuclei in putamen (+58%). Mice running for 45 min do not show any change in the labelling of nerve cell nuclei compared with mice moving freely in the cage. The [(3)H]uridine uptake in other cell types, e.g. other neurons in cortical layer 5, neurons in sensory cortex and in the other cell layers in motor cortex, were not different from control mice. We conclude that RNA synthesis is normally low in adult mouse brain, but that physical exercise stimulates RNA synthesis in specific populations of large neurons in the motor system. PMID- 17121642 TI - Maximum physical capacity testing in cancer patients undergoing chemotherapy: qualitative findings from an exercise program. AB - BACKGROUND: Over the past few years there has been a growing interest in the field of physical exercise in rehabilitation of cancer patients, leading to requirements for objective maximum physical capacity measurement (maximum oxygen uptake (VO(2max)) and one-repetition maximum (1RM)) to determine dose-response levels in different cancer diagnoses. AIM: To explore the patients' experiences of maximum physical capacity testing while concurrently undergoing chemotherapy and participating in a 6-week, 9 h weekly multidimensional exercise program. DESIGN AND METHOD: Prospective, exploratory study using semi-structured qualitative interviews conducted prior to and at termination of the program. The study included 100 patients (18-65 years, median 42 years) with or without residual disease and with mixed diagnoses. RESULTS: Following the intervention, cancer patients felt significantly safer in performing maximum physical capacity tests as these motivated them through self-perceived competitiveness and set a standard that served to encourage peak performance. CONCLUSION: The positive attitudes in this sample towards maximum physical capacity open the possibility of introducing physical testing early in the treatment process. However, the patients were self-referred and thus highly motivated and as such are not necessarily representative of the whole population of cancer patients treated with chemotherapy. PMID- 17121643 TI - Psychological impact of sports activity in spinal cord injury patients. AB - OBJECTIVE: To investigate whether sports activity is associated with better psychological profiles in patients with spinal cord injury (SCI) and to evaluate the effect of demographic factors on psychological benefits. METHODS: The State Trait Anxiety Inventory, Form X2 (STAI-X2), the Eysenck Personality Questionnaire for extraversion (EPQ-R (E)) and the questionnaire for depression (QD) were administered in a cross-sectional study of 137 males with spinal cord injury including 52 tetraplegics and 85 paraplegics. The subjects were divided into two groups according to sports activity participation (high frequency vs no sports participation). Moreover, multiple regression analysis was adopted to investigate the influence of demographic variables, such as age, educational level, occupational status and marital status, on psychological variables. RESULTS: Analysis of variance revealed significant differences among the groups for anxiety (STAI-X2), extraversion (EPQ-R (E)) and depression (QD). In particular, SCI patients who did not practice sports showed higher anxiety and depression scores and lower extraversion scores than sports participants. In addition, with respect to the paraplegics, the tetraplegic group showed the lowest depression scores. Following multiple regression analysis, only the sports activity factor remained as an independent factor of anxiety scores. CONCLUSION: These findings demonstrate that sports activity is associated with better psychological status in SCI patients, irrespective of tetraplegia and paraplegia, and that psychological benefits are not emphasized by demographic factors. PMID- 17121644 TI - Physical fitness profile of elite athletes with intellectual disability. AB - The aim of this study was to investigate the physical fitness profile of high performance athletes with intellectual disability (ID) in comparison with able bodied individuals. METHODS: Participants were 231 male and 82 female athletes. All evaluations were done using the EUROFIT physical fitness test. RESULTS: In comparison with population data, both male and female athletes with ID score better for flexibility and upper body muscle endurance, but have similar or lower values for running speed, speed of limb movement, and strength measures. Compared with age-matched physical education students, male athletes with ID score better for running speed and flexibility, and worse for strength. Female athletes with ID score not different from able-bodied individuals for flexibility, running speed, and upper body muscle endurance, but worse for strength measures. Athletes with ID also have poorer cardio respiratory endurance capacity compared with sportive peers without ID. Furthermore, male athletes have a more differentiated profile depending upon their sports discipline, compared with female athletes. CONCLUSION: It can be concluded that high-performance athletes with ID reach physical fitness levels that are equal to or lower than those of able-bodied sportive counterparts. Further research should investigate the importance of reduced muscle strength to be the limiting factor. PMID- 17121645 TI - Injury pattern in youth team handball: a comparison of two prospective registration methods. AB - The purpose of this study was to examine the injury incidence and pattern of injuries in youth female and male team handball players using two different prospective registration methods; match reports (90 teams, 1080 players) and coach reports (34 teams, 428 players). A total of 118 injuries were recorded by the coach report, of which 93 (79%) were acute injuries (incidence training: 0.9+/-0.16 injuries/1000 player hours; matches: 9.9+/-1.26; rate ratio vs training: 10.8 [95% confidence interval (CI) 7.0-16.6]; P<0.0001) and 25 (21%) were overuse injuries. Knee (26%) and ankle (24%) injuries accounted for half of the acute injuries (training: 0.5+/-0.12 injuries/1000/h; matches: 4.4+/-0.84; rate ratio vs training: 8.0 (95% CI 4.5-14.5); P<0.0001). No gender difference was found in the injury rate (rate ratio female vs male: 1.3 (95% CI 0.8-2.1); P=0.40). Most of the injuries occurred in the attacking phase by back or wing players doing a plant-and-cut, landing or turning movement, and more than half in contact situations with the opponent. Similar results were observed for acute match injuries in the match report. These results indicate that the rate of injuries in youth team handball is as high as at the senior level, and prevention should focus on knee and ankle injuries. The coach report seems to be the best method to register injuries in youth team handball to provide a full spectrum of injuries according to their type, incidence and severity. PMID- 17121646 TI - Injuries and preventive actions in elite Swedish volleyball. AB - The purpose of this study was to examine the prevalence of injury and the extent of preventive actions in elite Swedish volleyball players. Injuries to players in the elite male and female Swedish division, during the 2002-2003 season, were registered by using a questionnaire. Of the 158 volleyball players (70% response rate), a total of 82 players (52%) reported 121 injuries, during a total exposure time of 24 632 h, representing an overall incidence of 0.77 injuries per player. The majority of the injuries were located in the ankle (23%), followed by the knee (18%) and the back (15%). Most injuries (62%) were classified as being of minor severity. Most injuries occurred during training (47%), and 41% of the injuries had a gradual onset. Fifty-four percent of the injuries that could be related to a specific court situation occurred during blocking, and 30% during spiking. Most players (96%) participated in injury prevention training of some kind, generally performed without supervision (58%). Although most players took part in some kind of preventive action, one out of two players incurred an injury during the season, which indicates that the risk of suffering an injury in elite volleyball is relatively high. PMID- 17121647 TI - Who chooses anterior cruciate ligament reconstruction and why? A 2-year prospective study. AB - The purpose of this study was to investigate possible differences in age, gender, Tegner activity level prior to injury, knee joint laxity and the variables evaluated with knee osteoarthritis outcome score (KOOS) between patients who chose non-operative treatment (NT), early reconstruction (ER) and late reconstruction (LR). A second aim was to study whether patients who choose ER choose surgical treatment for the same reasons as patients who choose LR. Seventy two patients with an acute (< 1 month) anterior cruciate ligament (ACL) tear, who had not yet decided to have surgery, were included in this study. They filled out the Tegner activity score, assessing preinjury activity level, KOOS and underwent KT-1000 arthrometer examination. The 36 subjects who later chose reconstruction were questioned about reasons for their decision. A high preinjury activity level was associated with the choice of ACL reconstruction, but the choice of treatment was not associated with age, gender or the outcome variables measured with KOOS or KT-1000. Fifteen out of the 20 (75%) subjects who chose ER based their decision on assumptions of future problems and not on experience of knee function. Fourteen out of 16 (88%) subjects, who chose reconstructive surgery later, based their decision on experience of knee function. PMID- 17121648 TI - Relationships between physical activity and physical capacity in adolescent females and bone mass in adulthood. AB - This study investigates whether physical activity and physical performance in adolescence are positively related to adult bone mineral density (BMD). In 1974, physical activity, endurance, and muscular strength were measured in 204 randomly selected female students, age 16.1 +/- 0.3 year (range 15-17 years). Twenty years later, 36 of the women volunteered to undergo a measurement of their BMD. Women who were members in a sports club in adolescence had significantly higher adult BMD (mean differences of 5% to 17% depending on site) compared with subjects who were not engaged in a sports club. Furthermore, women with persistent weight bearing activity in adulthood had significantly higher BMD compared with women who had stopped being active or had never been active. The differences ranged between 5% and 19% with the highest difference found in trochanter BMD. Stepwise regression analyses showed that membership in a sports club at baseline was a significant independent predictor of BMD in the total body, lumbar spine, legs, trochanter, and femoral neck, explaining 17-26% of the variation in BMD. Change in body weight was a strong independent predictor of BMD of the total body and arms, explaining 8% of the variation in both sites. In addition, running performance at baseline was an independent predictor of total body BMD, whereas the two-hand lift performance significantly predicted BMD of the total body, legs and trochanter. The hanging leg-lift and handgrip were both significant predictors of arm BMD. In conclusion, membership in a sports club and site specific physical performance in adolescence together with the change in body weight were significantly associated with adult BMD in premenopausal women. PMID- 17121649 TI - A capsular incision leads to a fast osteoarthritic response, but also elevated levels of activated osteogenic protein-1 in rabbit knee joint cartilage. AB - We studied whether a small capsular incision alone, or combined with meniscectomy could induce early osteoarthritic changes in the rabbit knee. Thirty-one rabbits were operated on with a capsular incision in the left knee and meniscectomy in the right knee. Another 12 rabbits were used as controls. The rabbits were killed 3, 6 and 12 weeks after surgery. Osteoarthritic changes in the articular cartilage were evaluated by the modified Mankin score. The subchondral bone was evaluated by scintimetry ((99m)Tc-HDP) and semiquantitative grading of histological changes. Osteogenic protein (OP-1) in its mature and pro-form was examined by immunohistochemistry. Both a capsular incision and meniscectomy induced articular cartilage fibrillation and increased bone metabolic activity during the initial weeks after surgery. Capsular incision led to lesser changes than meniscectomy. Mature OP-1 was elevated, and its pro-form reduced, in meniscectomized knees. A similar pattern was observed in knees with capsular incision. Already 3 weeks after surgery, the articular cartilage and subchondral bone showed typical signs of early osteoarthritis (OA), and a reparative response was suggested by increased intensity of OP-1 staining. As these signs were also found in knees with capsular incision only, it appears that trauma-related factors such as increased bleeding and inflammation are critical for the development of OA. PMID- 17121650 TI - Tendinopathy and Doppler activity: the vascular response of the Achilles tendon to exercise. AB - BACKGROUND: Intratendinous Doppler activity has been interpreted as an equivalent of neovessels in the Achilles tendon and as a sign of tendinosis (AT). AIM: To evaluate the vascular response as indicated by color Doppler activity after repeated loading of both symptomatic and non-symptomatic Achilles tendons. MATERIAL AND METHODS: Ten non-trained, healthy subjects ran 5 km. Ultrasound (US) Doppler activity was determined before and after the exercise. Eleven patients with chronic AT performed 3 x 15 heavy-load eccentric exercise. The Achilles tendons were scanned before and immediately after the exercise. RESULTS: Non symptomatic: six Achilles tendons in five subjects had intratendinous Doppler activity before the exercise. All but two subjects (80%) had intratendinous Doppler activity after running. Symptomatic: all patients had Doppler activity in the tendons, with a median color fraction before eccentric exercise of 0.05 (range 0.01-0.33). The Doppler activity did not disappear after exercise. Tendons with a color fraction below the median at baseline increased significantly after the exercise (P=0.02). CONCLUSION: The mere presence of Doppler in the Achilles tendon does not per se indicate disease. Eccentric exercise does not extinguish the flow during or after one training session in patients with chronic AT. PMID- 17121654 TI - Relationship between seminal ascorbic acid and sperm DNA integrity in infertile men. AB - Ascorbic acid has recently been reported to protect sperm DNA from the damage induced by exogenous oxidative stress in vitro. But, there is no report on seminal ascorbic acid and sperm DNA fragmentation in infertile men. In this study, we asked whether sperm DNA damage correlates with seminal ascorbic acid levels. Sperm DNA fragmentation index (DFI) was analysed in 75 men by flow cytometry after acridine orange staining. We also measured the levels of seminal plasma ascorbic acid and total antioxidant capacity. Abnormal sperm DNA integrity (DFI >or= 30%) was observed in 12% of the patients with normal semen parameters and in 52% of the patients with abnormal semen parameters. There were significant correlations between the level of DFI and conventional semen parameters including sperm count, motility and morphology (r = -0.29, -0.55 and -0.53 respectively; p < 0.05). Seminal ascorbic acid level was significantly lower in the patients with leucospermia than the patient with normal semen parameters. Interestingly, a significantly greater percentage of men with abnormal DFI were observed in the patients with low levels of seminal ascorbic acid compared with those with normal or high levels of ascorbic acid (59% vs. 33%, p < 0.05). Men with insufficient seminal ascorbic acid frequently have sperm DNA damage. PMID- 17121655 TI - Effects of lithium carbonate on rat seminiferous tubules: an ultrastructural study. AB - Lithium salts are commonly used for treatment of bipolar disorder but prolonged treatment with therapeutic doses induces substantial toxic effects. In the present study we examined the effects of lithium carbonate on the ultrastructure of rat seminiferous tubules. Rats were exposed to lithium carbonate at doses of 35 mg/kg/day for 21 days. After lithium treatment, the tunica propria widened and folded together with convolutions of the basement membrane, myoid cells and lymphatic endothelium. In the seminiferous epithelium loss of germ cell attachment and appearance of expanded intercellular spaces between spermatogenic cells were observed. Early stages of spermatogenic cells showed nuclear protrusions or swellings because of an extensive enlargement of the outer nuclear membrane. Round spermatids exhibited abnormally shaped acrosomes and dilation of the subacrosomal space. Many abnormal, degenerated late spermatids with random orientation were seen towards the basal and adluminal compartments of the seminiferous epithelium. In addition, spermatids exhibited alteration in F-actin bundle ectoplasmic specialization and contained many mitochondria-associated granular bodies. PMID- 17121656 TI - Differences in SCSA outcome among boars with different sperm freezability. AB - Spermatozoa from some boars sustain the process of cryopreservation poorly and yield poor fertility after artificial insemination. Poor freezability has not been disclosed using conventional semen analyses. A defective chromatin can, if present in a substantial number of spermatozoa, affect the fertilizing ability of spermatozoa. Here we tested the hypothesis that nuclear DNA instability could explain differences in freezability among boars, and complement or supersede conventional tests for sperm quality such as sperm motility or membrane assessments. Frozen-thawed (FT) spermatozoa from a total of 44 stud boars were assessed by the sperm chromatin structure assay (SCSA), in relation to computer assisted sperm analysis-derived sperm motility variables and sperm viability (triple fluorescent microscopic staining), including three experiments. The first trial, including 24 boars, evaluated the relationship between the sperm motility and viability with levels of DNA integrity. The SCSA showed that most spermatozoa had intact DNA [levels of DNA fragmentation index (%DFI) ranging from 0.63% to 11.85%] significantly correlated (albeit weakly) with current sperm quality variables. The second trial, on 15 boars, assessed the influence of two different thawing rates (20 s at 37 degrees C vs. 8 s at 70 degrees C) and the post-thaw incubation times (0, 30, 150 and 300 min) at 37 degrees C on FT-boar sperm quality. The highest sperm survival (p < 0.05) and the lowest DNA damage (p < 0.01) were achieved when thawing was carried out at 70 degrees C for 8 s, without any change during the first 150 min of incubation. Finally, the third experiment studied if differences in sperm freezability showed by stud boar semen, as 'good' or 'bad' freezers by conventional analyses, could be attributed to differences in chromatin structure. All SCSA parameters were low, but significantly (p < 0.05 0.001) higher for 'bad' freezers, showing they had less homogeneous sperm chromatin than the 'good' freezers. The results indicate that SCSA outcome complements conventional assessment of FT-boar spermatozoa, disclosing differences in their ability to sustain freezing and thawing. However, the low overall DNA damage observed in FT spermatozoa seems to have poor biological significance. PMID- 17121657 TI - Alleviation of pre-exposure of mouse brain with low-dose 12C6+ ion or 60Co gamma ray on male reproductive endocrine damages induced by subsequent high-dose irradiation. AB - Irradiation has been widely reported to damage organisms by attacking on proteins, nucleic acid and lipids in cells. However, radiation hormesis after low dose irradiation has become the focus of research in radiobiology in recent years. To investigate the effects of pre-exposure of mouse brain with low-dose (12)C6+ ion or 60Co gamma (gamma)-ray on male reproductive endocrine capacity induced by subsequent high-dose irradiation, the brains of the B6C3F1 hybrid strain male mice were irradiated with 0.05 Gy of (12)C6+ ion or 60Co gamma-ray as the pre-exposure dose, and were then irradiated with 2 Gy as challenging irradiation dose at 4 h after pre-exposure. Serum pituitary gonadotropin hormones, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), testosterone, testis weight, sperm count and shape were measured on the 35th day after irradiation. The results showed that there was a significant reduction in the levels of serum FSH, LH, testosterone, testis weight and sperm count, and a significant increase in sperm abnormalities by irradiation of the mouse brain with 2 Gy of (12)C6+ ion or 60Co gamma-ray. Moreover, the effects were more obvious in the group irradiated by (12)C6+ ion than in that irradiated by 60Co gamma-ray. Pre-exposure with low-dose (12)C6+ ion or 60Co gamma-ray significantly alleviated the harmful effects induced by a subsequent high-dose irradiation. PMID- 17121658 TI - Early exposure to smoking and future fecundity among Danish twins. AB - Several studies have examined the effect of in utero exposure to smoking and fecundity among the offspring but the findings are contradictory. We therefore studied the waiting time to first pregnancy (TTP) and exposure to smoking in utero and childhood among Danish twins born between 1931 and 1952. Information about TTP, exposure to mothers smoking in pregnancy, exposure to smoking in childhood and current smoking among the male twins and smoking in their own pregnancy among female twins was collected by interview. Fecundability odds ratio (FOR) estimating the odds of conception in a cycle among exposed compared to the unexposed were calculated separately for female and male twins. A total of 1653 female and 1598 male twins reported a TTP. Female twins, exposed in utero, had reduced fecundability after control for confounders (FOR = 0.81; 95% CI 0.67 0.99). A nonsignificant increase in fecundity among male twins exposed to smoking in utero was found (FOR = 1.12; 95% CI 0.89-1.40). Among dizygotic twins of opposite sex sharing the same in utero exposures, the future fecundity of the male twin was unaffected by in utero exposure (FOR = 0.97; 95% CI 0.60-1.55) whereas the female twin had reduced fecundity (FOR = 0.65; 95% CI 0.47-0.91). This study supports that smoking is hazardous to the female fetus not only in the short term but also affects her future ability to conceive and makes it even more important to advise pregnant women to stop smoking. PMID- 17121659 TI - Alterations of the USP26 gene in Caucasian men. AB - The Ubiquitin Specific Protease 26 gene is a testis-specific gene that is located on the X chromosome. Sequence variants of this gene were previously reported in men with azoospermia caused by defects at the level of spermatogenesis. Especially a cluster of three changes (c.370_371insACA, c.494T>C and c.1423C>T) was frequently observed. To further define the role of this cluster of sequence variants in the USP26 gene, we have now analysed 202 control samples and 146 patients of Caucasian origin with cryptozoospermia or oligozoospermia. The detection method was based on a restriction reaction, by which the change c.494T>C can be detected. In none of the patients, the change c.494T>C was observed. Only in one man with normal spermatogenesis this sequence variant was detected. Sequencing can confirm the presence of the three changes of the USP26 gene. These data indicate that the cluster of changes is not restricted to men with severe testicular dysfunction. PMID- 17121663 TI - The mechanisms of atrial fibrillation. AB - In this article we have reviewed the mechanisms of atrial fibrillation (AF) with special emphasis on the thoracic veins. Based on a number of features, the thoracic veins are highly arrhythmogenic. The pulmonary vein (PV)-left atrial (LA) junction has discontinuous myocardial fibers separated by fibrotic tissues. The PV muscle sleeve is highly anisotropic. The vein of Marshall (VOM) in humans has multiple small muscle bundles separated by fibrosis and fat. Insulated muscle fibers can promote reentrant excitation, automaticity, and triggered activity. The PV muscle sleeves contain periodic acid-Schiff (PAS)-positive large pale cells that are morphologically reminiscent of Purkinje cells. These special cells could be the sources of focal discharge. Antiarrhythmic drugs have significant effects on PV muscle sleeves both at baseline and during AF. Both class I and III drugs have effects on wavefront traveling from PV to LA and from LA to PV. Separating the thoracic veins and the LA with ablation techniques also prevents PV-LA interaction. By reducing PV-LA interaction, pharmacological therapy and PV isolation reduce the activation rate in PV, intracellular calcium accumulation, and triggered activity. Therefore, thoracic vein isolation is an important technique in AF control. We conclude that thoracic veins are important in the generation and maintenance of AF. PMID- 17121672 TI - Possible criteria for inpatient psychiatric admissions: which patients are transferred from emergency services to inpatient psychiatric treatment? AB - BACKGROUND: Patients with psychiatric problems often seek help and assistance in hospital emergency departments. An important task of emergency room staff is to decide whether such patients need to be admitted or whether they can be treated on an outpatient basis. METHODS: Psychiatric treatments given in the Central Interdisciplinary Emergency Department (CED) at the Medical University of Hannover (MHH) in 2002 were analysed. RESULTS: Of a total of 2632 patients seeking psychiatric help, 51.4% were admitted for inpatient treatment. Patients with dementia syndromes were admitted more frequently than patients with other psychiatric diseases. Suicidality was often the reason for admission. Accompanied patients were less likely to be hospitalised, unless a care-order was in force. Restraining measures and acute medication also had an impact on the rate of admissions. CONCLUSION: The results may help psychiatrists in the emergency department to make a more effective decision regarding inpatient admission in the interest of the individual patient. PMID- 17121673 TI - A rapidly evolving secretome builds and patterns a sea shell. AB - BACKGROUND: Instructions to fabricate mineralized structures with distinct nanoscale architectures, such as seashells and coral and vertebrate skeletons, are encoded in the genomes of a wide variety of animals. In mollusks, the mantle is responsible for the extracellular production of the shell, directing the ordered biomineralization of CaCO3 and the deposition of architectural and color patterns. The evolutionary origins of the ability to synthesize calcified structures across various metazoan taxa remain obscure, with only a small number of protein families identified from molluskan shells. The recent sequencing of a wide range of metazoan genomes coupled with the analysis of gene expression in non-model animals has allowed us to investigate the evolution and process of biomineralization in gastropod mollusks. RESULTS: Here we show that over 25% of the genes expressed in the mantle of the vetigastropod Haliotis asinina encode secreted proteins, indicating that hundreds of proteins are likely to be contributing to shell fabrication and patterning. Almost 85% of the secretome encodes novel proteins; remarkably, only 19% of these have identifiable homologues in the full genome of the patellogastropod Lottia scutum. The spatial expression profiles of mantle genes that belong to the secretome is restricted to discrete mantle zones, with each zone responsible for the fabrication of one of the structural layers of the shell. Patterned expression of a subset of genes along the length of the mantle is indicative of roles in shell ornamentation. For example, Has-sometsuke maps precisely to pigmentation patterns in the shell, providing the first case of a gene product to be involved in molluskan shell pigmentation. We also describe the expression of two novel genes involved in nacre (mother of pearl) deposition. CONCLUSION: The unexpected complexity and evolvability of this secretome and the modular design of the molluskan mantle enables diversification of shell strength and design, and as such must contribute to the variety of adaptive architectures and colors found in mollusk shells. The composition of this novel mantle-specific secretome suggests that there are significant molecular differences in the ways in which gastropods synthesize their shells. PMID- 17121674 TI - Validation of the Portuguese version of the Lithium Attitudes Questionnaire (LAQ) in bipolar patients treated with lithium: cross-over study. AB - BACKGROUND: Poor adherence to lithium is very common in bipolar patients and it is a frequent cause of recurrence during prophylactic treatment. Several reports suggest that attitudes of bipolar patients interfere with adherence to lithium. The Lithium Attitudes Questionnaire (LAQ) is a brief questionnaire developed as a means of identifying and grouping the problems patients commonly have with taking lithium regularly. The original version is validated in patients, but a validated version in Portuguese is not yet available. METHODS: One-hundred six patients with bipolar disorder (DSM-IV criteria) criteria under lithium treatment for at least one month were assessed using LAQ. LAQ is a brief questionnaire administered under interview conditions, which includes 19 items rating attitudes towards prophylactic lithium treatment. We analysed the internal consistency, concurrent validity, sensitivity and specificity of the Portuguese version of LAQ. RESULTS: The internal consistency, evaluated by Cronbach's alpha was 0.78. The mean total LAQ score was 4.1. Concurrent validity was confirmed by a negative correlation between plasma lithium concentration and total LAQ score (r = -0,198; p = 0.048). We analysed the scale's discriminative capacity revealing a sensitivity of 69% and a specificity of 71% in the identification of negative attitudes of bipolar patients. CONCLUSION: The psychometric assessment of the Portuguese version of LAQ showed good internal consistency, sensitivity and specificity. The results were similar to the original version in relation to attitudes of bipolar patients towards lithium therapy. PMID- 17121675 TI - Context-sensitive autoassociative memories as expert systems in medical diagnosis. AB - BACKGROUND: The complexity of our contemporary medical practice has impelled the development of different decision-support aids based on artificial intelligence and neural networks. Distributed associative memories are neural network models that fit perfectly well to the vision of cognition emerging from current neurosciences. METHODS: We present the context-dependent autoassociative memory model. The sets of diseases and symptoms are mapped onto a pair of basis of orthogonal vectors. A matrix memory stores the associations between the signs and symptoms, and their corresponding diseases. A minimal numerical example is presented to show how to instruct the memory and how the system works. In order to provide a quick appreciation of the validity of the model and its potential clinical relevance we implemented an application with real data. A memory was trained with published data of neonates with suspected late-onset sepsis in a neonatal intensive care unit (NICU). A set of personal clinical observations was used as a test set to evaluate the capacity of the model to discriminate between septic and non-septic neonates on the basis of clinical and laboratory findings. RESULTS: We show here that matrix memory models with associations modulated by context can perform automatic medical diagnosis. The sequential availability of new information over time makes the system progress in a narrowing process that reduces the range of diagnostic possibilities. At each step the system provides a probabilistic map of the different possible diagnoses to that moment. The system can incorporate the clinical experience, building in that way a representative database of historical data that captures geo-demographical differences between patient populations. The trained model succeeds in diagnosing late-onset sepsis within the test set of infants in the NICU: sensitivity 100%; specificity 80%; percentage of true positives 91%; percentage of true negatives 100%; accuracy (true positives plus true negatives over the totality of patients) 93,3%; and Cohen's kappa index 0,84. CONCLUSION: Context-dependent associative memories can operate as medical expert systems. The model is presented in a simple and tutorial way to encourage straightforward implementations by medical groups. An application with real data, presented as a primary evaluation of the validity and potentiality of the model in medical diagnosis, shows that the model is a highly promising alternative in the development of accuracy diagnostic tools. PMID- 17121676 TI - Serine phosphorylation regulates paxillin turnover during cell migration. AB - BACKGROUND: Paxillin acts as an adaptor protein that localizes to focal adhesion. This protein is regulated during cell migration by phosphorylation on tyrosine, serine and threonine residues. Most of these phosphorylations have been implicated in the regulation of different steps of cell migration. The two major phosphorylation sites of paxillin in response to adhesion to an extracellular matrix are serines 188 and 190. However, the function of this phosphorylation event remains unknown. The purpose of this work was to determine the role of paxillin phosphorylation on residues S188 and S190 in the regulation of cell migration. RESULTS: We used NBT-II epithelial cells that can be induced to migrate when plated on collagen. To examine the role of paxillin serines 188/190 in cell migration, we constructed an EGFP-tagged paxillin mutant in which S188/S190 were mutated into unphosphorylatable alanine residues. We provide evidence that paxillin is regulated by proteasomal degradation following polyubiquitylation of the protein. During active cell migration on collagen, paxillin is protected from proteasome-dependent degradation. We demonstrate that phosphorylation of serines 188/190 is necessary for the protective effect of collagen. In an effort to understand the physiological relevance of paxillin protection from degradation, we show that cells expressing the paxillin S188/190A interfering mutant spread less, have reduced protrusive activity but migrate more actively. CONCLUSION: Our data demonstrate for the first time that serine regulated degradation of paxillin plays a key role in the modulation of membrane dynamics and consequently, in the control of cell motility. PMID- 17121677 TI - Exploring glycopeptide-resistance in Staphylococcus aureus: a combined proteomics and transcriptomics approach for the identification of resistance-related markers. AB - BACKGROUND: To unravel molecular targets involved in glycopeptide resistance, three isogenic strains of Staphylococcus aureus with different susceptibility levels to vancomycin or teicoplanin were subjected to whole-genome microarray based transcription and quantitative proteomic profiling. Quantitative proteomics performed on membrane extracts showed exquisite inter-experimental reproducibility permitting the identification and relative quantification of >30% of the predicted S. aureus proteome. RESULTS: In the absence of antibiotic selection pressure, comparison of stable resistant and susceptible strains revealed 94 differentially expressed genes and 178 proteins. As expected, only partial correlation was obtained between transcriptomic and proteomic results during stationary-phase. Application of massively parallel methods identified one third of the complete proteome, a majority of which was only predicted based on genome sequencing, but never identified to date. Several over-expressed genes represent previously reported targets, while series of genes and proteins possibly involved in the glycopeptide resistance mechanism were discovered here, including regulators, global regulator attenuator, hyper-mutability factor or hypothetical proteins. Gene expression of these markers was confirmed in a collection of genetically unrelated strains showing altered susceptibility to glycopeptides. CONCLUSION: Our proteome and transcriptome analyses have been performed during stationary-phase of growth on isogenic strains showing susceptibility or intermediate level of resistance against glycopeptides. Altered susceptibility had emerged spontaneously after infection with a sensitive parental strain, thus not selected in vitro. This combined analysis allows the identification of hundreds of proteins considered, so far as hypothetical protein. In addition, this study provides not only a global picture of transcription and expression adaptations during a complex antibiotic resistance mechanism but also unravels potential drug targets or markers that are constitutively expressed by resistant strains regardless of their genetic background, amenable to be used as diagnostic targets. PMID- 17121678 TI - Impact of concomitant DMARD therapy on adherence to treatment with etanercept and infliximab in rheumatoid arthritis. Results from a six-year observational study in southern Sweden. AB - The objective of this work is to compare the adherence to therapy of patients receiving etanercept and infliximab during first tumour necrosis factor (TNF) blocking treatment course in rheumatoid arthritis. Special emphasis is placed on potential predictors for treatment termination and the impact of concomitant methotrexate (MTX) or other disease-modifying antirheumatic drugs (DMARDs). Patients (n = 1,161) with active rheumatoid arthritis, not responding to at least two DMARDs including MTX starting etanercept or infliximab therapy for the first time, were included in a structured clinical follow-up protocol. Information on diagnosis, disease duration, previous and ongoing DMARDs, treatment start and termination, as well as cause of withdrawal was prospectively collected during the period of March 1999 through December 2004. Patients were divided into six groups according to TNF-blocking drugs and concomitant DMARDs. Five-year level (one-year) of adherence to therapy was 36% (69%) for patients receiving infliximab in combination with MTX compared with 65% (89%) for patients treated with etanercept and MTX (p < 0.001). Cox regression models showed that the risk for premature treatment termination of patients treated with infliximab was threefold higher than for etanercept (p < 0.001). Also, the regression analysis showed that patients receiving concomitant MTX had better treatment continuation than patients treated solely with TNF blockers (p < 0.001). Moreover, patients receiving concomitant MTX had superior drug survival than patients receiving other concomitant DMARDs (p < 0.010). The superior effect of MTX was associated primarily with fewer treatment terminations because of adverse events. In addition, the study identifies low C-reactive protein level, high age, elevated health assessment questionnaire score, and higher previous number of DMARDs as predictors of premature treatment termination. In summary, treatment with etanercept has higher adherence to therapy than treatment with infliximab. Concomitant MTX is associated with improved treatment continuation of biologics when compared with both TNF blockers as monotherapy and TNF blockers combined with other DMARDs. PMID- 17121679 TI - A fungal phylogeny based on 42 complete genomes derived from supertree and combined gene analysis. AB - BACKGROUND: To date, most fungal phylogenies have been derived from single gene comparisons, or from concatenated alignments of a small number of genes. The increase in fungal genome sequencing presents an opportunity to reconstruct evolutionary events using entire genomes. As a tool for future comparative, phylogenomic and phylogenetic studies, we used both supertrees and concatenated alignments to infer relationships between 42 species of fungi for which complete genome sequences are available. RESULTS: A dataset of 345,829 genes was extracted from 42 publicly available fungal genomes. Supertree methods were employed to derive phylogenies from 4,805 single gene families. We found that the average consensus supertree method may suffer from long-branch attraction artifacts, while matrix representation with parsimony (MRP) appears to be immune from these. A genome phylogeny was also reconstructed from a concatenated alignment of 153 universally distributed orthologs. Our MRP supertree and concatenated phylogeny are highly congruent. Within the Ascomycota, the sub-phyla Pezizomycotina and Saccharomycotina were resolved. Both phylogenies infer that the Leotiomycetes are the closest sister group to the Sordariomycetes. There is some ambiguity regarding the placement of Stagonospora nodurum, the sole member of the class Dothideomycetes present in the dataset. Within the Saccharomycotina, a monophyletic clade containing organisms that translate CTG as serine instead of leucine is evident. There is also strong support for two groups within the CTG clade, one containing the fully sexual species Candida lusitaniae, Candida guilliermondii and Debaryomyces hansenii, and the second group containing Candida albicans, Candida dubliniensis, Candida tropicalis, Candida parapsilosis and Lodderomyces elongisporus. The second major clade within the Saccharomycotina contains species whose genomes have undergone a whole genome duplication (WGD), and their close relatives. We could not confidently resolve whether Candida glabrata or Saccharomyces castellii lies at the base of the WGD clade. CONCLUSION: We have constructed robust phylogenies for fungi based on whole genome analysis. Overall, our phylogenies provide strong support for the classification of phyla, sub-phyla, classes and orders. We have resolved the relationship of the classes Leotiomyctes and Sordariomycetes, and have identified two classes within the CTG clade of the Saccharomycotina that may correlate with sexual status. PMID- 17121680 TI - Distinct cellular responses differentiating alcohol- and hepatitis C virus induced liver cirrhosis. AB - BACKGROUND: Little is known at the molecular level concerning the differences and/or similarities between alcohol and hepatitis C virus induced liver disease. Global transcriptional profiling using oligonucleotide microarrays was therefore performed on liver biopsies from patients with cirrhosis caused by either chronic alcohol consumption or chronic hepatitis C virus (HCV). RESULTS: Global gene expression patterns varied significantly depending upon etiology of liver disease, with a greater number of differentially regulated genes seen in HCV infected patients. Many of the gene expression changes specifically observed in HCV-infected cirrhotic livers were expectedly associated with activation of the innate antiviral immune response. We also compared severity (CTP class) of cirrhosis for each etiology and identified gene expression patterns that differentiated ethanol-induced cirrhosis by class. CTP class A ethanol-cirrhotic livers showed unique expression patterns for genes implicated in the inflammatory response, including those related to macrophage activation and migration, as well as lipid metabolism and oxidative stress genes. CONCLUSION: Stages of liver cirrhosis could be differentiated based on gene expression patterns in ethanol induced, but not HCV-induced, disease. In addition to genes specifically regulating the innate antiviral immune response, mechanisms responsible for differentiating chronic liver damage due to HCV or ethanol may be closely related to regulation of lipid metabolism and to effects of macrophage activation on deposition of extracellular matrix components. PMID- 17121681 TI - Predictors of gallstone composition in 1025 symptomatic gallstones from Northern Germany. AB - BACKGROUND: Gallstones represent a prevalent and costly health problem. The changing epidemiology and the emerging non-surgical interventions for gallstone disease necessitate the definition of target populations for future therapies. This study aimed to define patterns of gallstone composition and identify demographic predictors of gallstone composition in a large sample of symptomatic gallstones from Northern Germany. METHODS: One thousand and seventy-four post cholecystectomy gallstone specimens were obtained. Demographic and clinical information was provided by questionnaire (N = 1025 independent individuals with complete information). Two samples from each gallstone were analyzed using Fourier transformed infrared spectrometry. RESULTS: The most prevalent substance was cholesterol, which was detected in 95.0% of gallstone specimens. Bilirubin and bilirubinate were present in 30.0% and calcium was detected in 10.0% of the spectra. Ninety-two percent of measurements from the same stone yielded the same "main" substances, indicating a homogenous stone composition in most cases. Female sex and higher body mass index (BMI) were associated with the presence of cholesterol as a main substance in the gallstones (p < 0.001). CONCLUSION: The changing epidemiology of gallstone disease is reflected by a marked shift in stone composition: Only two percent of stones in this study were pigment stones as compared to 91% percent of stones containing cholesterol as a main substance. Obese individuals from Germany with a BMI > 30 kg/m2 have in 95% cholesterol dominant gallstones and represent a potential target population for non-surgical interventions for the prevention or treatment of cholesterol stones. PMID- 17121682 TI - Multilocus sequence typing method for identification and genotypic classification of pathogenic Leptospira species. AB - BACKGROUND: Leptospira are the parasitic bacterial organisms associated with a broad range of mammalian hosts and are responsible for severe cases of human Leptospirosis. The epidemiology of leptospirosis is complex and dynamic. Multiple serovars have been identified, each adapted to one or more animal hosts. Adaptation is a dynamic process that changes the spatial and temporal distribution of serovars and clinical manifestations in different hosts. Serotyping based on repertoire of surface antigens is an ambiguous and artificial system of classification of leptospiral agents. Molecular typing methods for the identification of pathogenic leptospires up to individual genome species level have been highly sought after since the decipherment of whole genome sequences. Only a few resources exist for microbial genotypic data based on individual techniques such as Multiple Locus Sequence Typing (MLST), but unfortunately no such databases are existent for leptospires. RESULTS: We for the first time report development of a robust MLST method for genotyping of Leptospira. Genotyping based on DNA sequence identity of 4 housekeeping genes and 2 candidate genes was analyzed in a set of 120 strains including 41 reference strains representing different geographical areas and from different sources. Of the six selected genes, adk, icdA and secY were significantly more variable whereas the LipL32 and LipL41 coding genes and the rrs2 gene were moderately variable. The phylogenetic tree clustered the isolates according to the genome-based species. CONCLUSION: The main advantages of MLST over other typing methods for leptospires include reproducibility, robustness, consistency and portability. The genetic relatedness of the leptospires can be better studied by the MLST approach and can be used for molecular epidemiological and evolutionary studies and population genetics. PMID- 17121683 TI - Clinical features and hormonal profiles of cloprostenol-induced early abortions in heifers monitored by ultrasonography. AB - BACKGROUND: The present study describes the clinical features and plasma profiles of bovine pregnancy-associated glycoprotein 1 (bPAG1), the main metabolite of prostaglandin F2alpha (PG metabolite) and progesterone (P4) in heifers in which early abortions were induced. METHODS: Early abortions were induced in four heifers with cloprostenol and monitored by ultrasonography. Blood samples were collected and the plasma were analyzed for bPAG 1, P4 and PG metabolite. RESULTS: The foetal heartbeat rates varied from 170-186 beats per minute for all foetuses up to the date of cloprostenol treatment. Foetal death was confirmed within two days after cloprostenol treatment. Prior to cloprostenol injection, blood plasma concentrations of bPAG1, PG metabolite and P4 varied from 8.4-40.0 ng/mL, 158-275 pmol/L and 20.7-46.9 nmol/L, respectively. After the foetus expelled, the plasma level of bPAG1 began to decrease but the decrease was small and gradual. The estimated half-life of bPAG1 was 1.8-6.6 days. The plasma level of the PG metabolite started to have short lasting peaks (above 300 pmol/L) within three hours after cloprostenol treatment. The plasma concentrations of P4 dropped sharply to less than 4 nmol/L after 24 hours of cloprostenol injection. CONCLUSION: The current findings indicated that after early closprostenol-induced foetal death, the plasma concentration of bPAG1 decreased gradually and showed a tendency of variation with the stages of pregnancy. PMID- 17121684 TI - Phospholipase A1 from Trypanosoma cruzi infective stages generates lipid messengers that activate host cell protein kinase c. AB - Here we have studied phospholipase A1 (Plase A1) from Trypanosoma cruzi infective stages and it's possible role regarding the interaction with mammalian host cells. Plase A1 was mainly detected as a membrane-bound activity in the infective amastigote and trypomastigote stages, being remarkably higher with respect to the non-infective epimastigotes. It is noteworthy that only the infective stages secreted Plase A1. Moreover, along the differentiation process from epimastigotes into metacyclic trypomastigotes, the secreted enzyme activity increased simultaneously with the appearance of metacyclic forms, as expected. Since this enzyme is predominantly membrane-associated and secreted by the infective stages, Vero cell lipid profile modifications were analysed after interaction with either intact infective parasites or purified T. cruzi Plase A1. Significant changes in Vero cell lipid composition were observed, with the appearance of free fatty acids, diacylglycerol and lysophosphatidylcholine. Concomitantly with the generation of second lipid messengers, host cell protein kinase C activation was demonstrated. These results indicate that T. cruzi Plase A1 could play a critical role in the early events of parasite-host cell interaction that precede invasion. PMID- 17121685 TI - Behaviour and chemical signature of pre-hibernating females of Polistes dominulus infected by the strepsipteran Xenos vesparum. AB - Polistes dominulus are social wasps which are the host of the strepsipteran endoparasite Xenos vesparum. In the hibernating phase, unparasitized and parasitized wasps leave natal nests and aggregate together in sheltered quarters. In aggregations, wasps are socially active, and some individuals perform helping behaviour. Here we investigated if castrated parasitized wasps perform worker tasks in mixed aggregations. Moreover, by gas chromatography and mass spectrometry, we examined the cuticular hydrocarbons of unparasitized and parasitized wasps to evaluate if the infection alters the composition of cuticular waxes that are recognition cues in social insects. In clusters, infected females do not perform helping behaviour and they are less active than unparasitized wasps. Cuticular hydrocarbons are slightly differentiated between unparasitized and parasitized wasps but, generally, unparasitized wasps are more similar to wasps infected by Xenos females compared to wasps infected by Xenos males. Wasps infected by Xenos males do not usually survive the winter. This chemical similarity is probably a consequence of the similar physiological condition of unparasitized and female-affected Polistes wasps. At this stage, it is difficult to affirm whether these modifications are a true parasite manipulation or a consequence of infection. PMID- 17121686 TI - Social inequalities in antidepressant treatment and mortality: a longitudinal register study. AB - BACKGROUND: Despite an increased prevalence of depression among people of low socio-economic position, it remains unclear whether their treatment with antidepressants appropriately matches their increased need compared with people from more affluent backgrounds. This study examined socio-economic differences in antidepressant prescriptions and mortality related to depressive disorders. METHOD: A longitudinal register study of 17947 male and 47458 female local government employees with linked information on socio-economic indicators (education and occupational status) and data on antidepressant use and mortality associated with depressive disorder (suicide, alcohol-related deaths) during the years 1994 to 2000. RESULTS: In men, antidepressant treatment was less common among low educational groups than among high educational groups (OR 0.87, 95% CI 0.76-0.99) and a corresponding difference was seen between occupational statuses (OR for manual v. upper non-manual 0.72, 95% CI 0.62-0.84). In women, socio economic position was not associated with antidepressant use. However, both among the men and women, employees with low socio-economic position had increased risk for mental-health-related mortality, as indicated by suicides, deaths from alcohol-related causes, and all-cause mortality. CONCLUSIONS: These data suggest a mismatch in the treatment of depression relative to apparent clinical need, with the lowest levels of treatment concentrated in the lower socio-economic groups, despite evidence of their increased prevalence of depression and suicide. PMID- 17121687 TI - Patterns of contraceptive use in Australia: analysis of the 2001 National Health Survey. AB - The purpose of this paper is to review the patterns of contraceptive use in Australia, using data from a nationally representative sample of 5872 women aged 18 to 49. This survey was conducted by the Australian Bureau of Statistics in 2001 as part of the National Health Survey. Results of the analysis indicate that the oral contraceptive pill and condom were the two most frequently used methods. More than 76% of the respondents reported having ever used the pill. Over 23% of women were currently using condoms; of these 80% of the condom users used them for contraception - this included 36% who used condoms for both protection against infection and for contraception - and the remainder used them only for protection. Withdrawal was the third most popular non-surgical method up to age 40. Few women used IUDs, injections or diaphragms. Just over 3% of the respondents were using natural methods with the highest rate reported among those in their 30s. The 'morning-after pill' was reported mostly by women aged 18-24; however, there was no evidence to suggest that it was being used as a primary method of birth control. Contraceptive use declined in older women who turned to sterilization for themselves and/or their partners. Use of the contraceptive pill was somewhat higher among better-educated women, but lower among less-educated women and those from non-English-speaking backgrounds. PMID- 17121688 TI - The sources of co-morbidity between major depression and generalized anxiety disorder in a Swedish national twin sample. AB - BACKGROUND: Prior studies report high levels of co-morbidity between major depression (MD) and generalized anxiety disorder (GAD) and suggest that these disorders are closely related genetically. The personality trait of neuroticism (N) is substantially correlated with risk for MD and GAD. METHOD: Bivariate twin models were applied to lifetime diagnoses of modified DSM-IV diagnosis of MD and GAD obtained at personal interview in 1998-2003 with 37296 twins from the population-based Swedish Twin Registry. A trivariate Cholesky model with N, MD and GAD was applied to a subset (23280 members of same-sex twin pairs) who completed a self-report questionnaire assessing N in 1972-1973. RESULTS: In the best-fit bivariate model, the genetic correlation between MD and GAD was estimated at +1.00 in females and +0.74 in males. Individual-specific environmental factors were also shared between the two disorders with an estimated correlation of +0.59 in males and +0.36 in females. In the best-fit trivariate Cholesky model, genetic factors indexed by N impacted equally on risk for MD and GAD in males and females. However, in both sexes, genetic risk factors indexed by N contributed only around 25% to the genetic correlation between MD and GAD. CONCLUSION: Genetic risk factors for lifetime MD and GAD are strongly correlated, with higher correlations in women than in men. Although genetic risk factors indexed by the personality trait of N contribute substantially to risk for both MD and GAD, the majority of genetic covariance between the two disorders results from factors not shared with N. PMID- 17121689 TI - A prospective study of dysfunctional thinking and the regulation of negative intrusive memories in bipolar 1 disorder: implications for affect regulation theory. AB - BACKGROUND: Cognitive behavioural therapy (CBT) has been implemented in bipolar (BP) disorder with varying degrees of success. The link between CBT for BP disorder and underlying theory is not clear. There have been attempts to identify a unique style of dysfunctional thinking in BP disorder analogous to that in unipolar (UP) disorder but this has suffered from a dearth of prospective studies controlling for mood and phase of illness. In this prospective study, we have examined whether dysfunctional thinking and the (dys)regulation of traumatic memories are trait vulnerability factors in BP versus UP mood disorders. METHOD: BP-1 and UP groups were followed-up from acute episode to recovery, and compared with a healthy control group. Measures of dysfunctional thinking and linked personality dimensions were taken, together with measures assessing autobiographical memory retrieval (AMT). The presence and control of intrusive memories of traumatic events in BP and UP disorder were assessed. RESULTS: Controlling for mood symptoms and phase of illness, the BP (but not the UP) group was largely indistinguishable from controls on dysfunctional thinking. Intrusive memories of traumatic events were present in 45% of the BP and 48% of the UP groups; and those without intrusions were more overgeneral on the AMT in all phases of illness. CONCLUSION: These findings are in line with the 'affect regulation' hypothesis for UP disorder and deliberate self-harm. This may help in understanding the mode of action of CBT in BP disorder and to further improve the therapy, primarily through the promotion of affective regulation, which is one component of the complex CBT intervention for BP disorder. PMID- 17121690 TI - Comparison of alternative models for personality disorders. AB - BACKGROUND: The categorical classification system for personality disorder (PD) has been frequently criticized and several alternative dimensional models have been proposed. METHOD: Antecedent, concurrent and predictive markers of construct validity were examined for three models of PDs: the Five-Factor Model (FFM), the Schedule for Nonadaptive and Adaptive Personality (SNAP) model and the DSM-IV in the Collaborative Study of Personality Disorders (CLPS) sample. RESULTS: All models showed substantial validity across a variety of marker variables over time. Dimensional models (including dimensionalized DSM-IV) consistently outperformed the conventional categorical diagnosis in predicting external variables, such as subsequent suicidal gestures and hospitalizations. FFM facets failed to improve upon the validity of higher-order factors upon cross validation. Data demonstrated the importance of both stable trait and dynamic psychopathological influences in predicting external criteria over time. CONCLUSIONS: The results support a dimensional representation of PDs that assesses both stable traits and dynamic processes. PMID- 17121691 TI - Multiply antibiotic-resistant Vibrio cholerae O1 biotype El Tor strains emerge during cholera outbreaks in Zambia. AB - Antibiotic resistance data, made available from laboratory records during eight cholera outbreaks between 1990 and 2004 showed Vibrio cholerae serogroup O1 to have a low level of resistance (2-3%) to tetracycline during 1990-1991. Resistance increased for tetracycline (95%), chloramphenicol (78%), doxycycline (70%) and trimethoprim-sulphamethoxazole (97%) in subsequent outbreaks. A significant drop in resistance to tetracycline and chloramphenicol followed the adoption of a national policy to replace tetracycline with erythromycin for treating cholera. Sixty-nine strains from cholera outbreaks in Zambia between 1996 and 2004, were examined for antibiotic resistance and basic molecular traits. A 140 MDa conjugative, multidrug-resistant plasmid was found to encode tetracycline resistance in strains from 1996/1997 whereas strains from 2003/2004 were resistant to furazolidone, but susceptible to tetracycline, and lacked this plasmid. PCR revealed 25 of 27 strains from 1996/1997 harboured the intl1 class 1 integron but lacked SXT, a conjugative transposon element. Similar screening of 42 strains from 2003/2004 revealed all carried SXT but not the intl1 class 1 integron. All 69 strains, except two, one lacking ctxA and the other rstR and thus presumably truncated in the CTX prophage region, were positive for important epidemic markers namely rfbO1, ctxA, rstR2, and tcpA of El Tor biotype. Effective cholera management is dependent on updated reports on culture and sensitivity to inform the choice of antibiotic. Since the emergence of antibiotic resistance may significantly influence strategies for controlling cholera, continuous monitoring of epidemic strains is crucial. PMID- 17121692 TI - Modelling outbreak control for pneumonic plague. AB - Although pneumonic plague is listed by the Centers of Disease Control in the leading 'critical biological agents', very few studies exist on this subject. In this study, a mathematical compartment model was used to describe the geographical and temporal spread of an epidemic of pneumonic plague following its use as a biological weapon. Univariate and multivariate analyses were performed in order to assess the key parameters for the control of an outbreak in France. If interventions were taken 10 days after an attack, a reference scenario of 1000 index cases in Paris would lead to 2500 deaths. The results of the study indicate that the rapidity of onset of interventions has the largest effect on the final size of the epidemic, followed by wearing masks, treating contacts preventively and quarantine. Limiting inter-regional mixing does little to reduce casualties, although it does confine them to a single region. PMID- 17121693 TI - Increased rainfall is associated with increased risk for legionellosis. AB - Legionnaires' disease (LD) is caused by Legionella species, most of which live in water. The Mid-Atlantic region experienced a sharp rise in LD in 2003 coinciding with a period of record-breaking rainfall. To investigate a possible relationship, we analysed the association between monthly legionellosis incidence and monthly rainfall totals from January 1990 to December 2003 in five Mid Atlantic states. Using negative binomial model a 1-cm increase in rainfall was associated with a 2.6% (RR 1.026, 95% CI 1.012-1.040) increase in legionellosis incidence. The average monthly rainfall from May to September 1990-2002 was 10.4 cm compared to 15.7 cm from May to September 2003. This change in rainfall corresponds to an increased risk for legionellosis of approximately 14.6% (RR 1.146, 95% CI 1.067-1.231). Legionellosis incidence increased during periods of increased rainfall; identification of mechanisms that increase exposure and transmission of Legionella during rainfall might lead to opportunities for prevention. PMID- 17121694 TI - Phase-contrast magnetic resonance angiography for assessment of intracranial venous sinus lesions. AB - OBJECTIVE: To review the presentations of intracranial venous sinus lesions in phase-contrast (PC) magnetic resonance angiography (MRA) and discuss the diagnostic value of this imaging modality for these lesions. METHODS: This study involved 52 patients with intracranial venous sinus lesions, including 21 with dural venous sinus malformation, 7 dural arteriovenous fistula (6 caroticocaveneous fistula, CCF), 5 venous sinus thrombus and 19 meningioma invading the venous sinuses. All patients underwent PC and time-of-flight (TOF) MRA with a 1.5 Tesla MR scanner, with 28 of them undergoing subsequent digital subtraction angiography (DSA). RESULTS: PC MRA showed extensive hyperintense signals that identified the lesions, feeding arteries, deferent veins and venous sinuses in 21 cases of dural arteriovenous fistula, but the lesion signals appeared homogeneous. In these cases, PC and TOF MRA had equally good performance in displaying the feeding arteries with success in 20 of the 21 cases (95.2%), whereas the former resulted in a greater rate of deferent vein display [90.5% (19/21)] than the latter [76.2% (16/21), P<0.05]. The 6 CCF cases showed ipsilateral cavernous sinus expansion and upper ophthalmic vein dilations. In the 5 cases of venous sinus thrombis, 4 had transverse sinus thrombus and 1 superior sagittal sinus thrombus. All the cases presented signal disappearance in the sinuses in PC MRA and hyperintense signals in MRI, and some cases had also cerebral superficial hemorrhage and edema. The 19 cases of meningioma invading the venous sinus displayed sinus shift, stenosis and occlusion in PC MRA. CONCLUSION: PC MRA has great advantages in displaying the anatomical structure and lesions of the venous sinuses. PMID- 17121695 TI - Protein expression of human neuron-specific enolase and its antiserum preparation. AB - OBJECTIVE: To clone and express human neuron-specific enolase (HuNSE) protein and prepare NSE-specific antibody for prion disease diagnosis. METHODS: HuNSE gene was amplified by RT-PCR and subcloned into a HIS-tagged expression vector pQE30 after sequence verification. HIS-NSE fusion protein expression was obtained in E. coli M15 after IPTG induction followed by purification of the fusion protein by Ni-NTA affinity chromatography. Two male rabbits were immunized for 4 times with the purified protein, and the antiserum against NSE protein was collected and evaluated by enzyme-linked immunosorbent assay (ELISA), Western blotting and immunohistochemistry. RESULTS: SDS-PAGE assay yielded an approximately 22 kD HIS NSE fusion protein. The prepared antiserum could recognize both recombinant NSE protein and native NSE protein extracted from the brain tissues of different mammalian species as shown by Western blotting and immunohistochemistry. CONCLUSION: High expression of HuNSE is obtained in E. coli and the prepared antiserum against HuNSE can be used potentially for diagnosis of prion-associated diseases and other nervous degeneration diseases. PMID- 17121696 TI - [cDNA microarray-based screening of differentially expressed genes in macrophages in the spleen of patients with portal hypertension and hypersplenism]. AB - OBJECTIVE: To identify the differentially expressed genes associated with hypersplenism in patients with portal hypertension. METHODS: The total RNA were extracted from the macrophages isolated from normal spleen and the spleen of patients with portal hypertension and reversely transcribed to cDNA with the incorporation of fluorescent (cy3 and cy5)-labeled dCTP to prepare the hybridization probes. After hybridization of Biostar-H140s chip containing 14,112 spots of cDNAs with the prepared probes, the gene chip was scanned for fluorescence intensity to screen the differently expressed genes. Three gene chips were used for hybridization and only the genes with differential expression in all the three chips were considered to associate with hypersplenism in patients with portal hypertension. RESULTS: Totaling 896, 1330 and 898 genes were identified to be differentially expressed by the three chips, respectively, and 121 genes (0.86%) showed differential expression in all the three chips, including 21 up-regulated known genes and 73 down-regulated known genes. The differently expressed genes were functionally related with ion channels and transport proteins, cyclins, cytoskeleton, cell receptors, cell signal transduction, metabolism, immunity, and so forth. These genes might be involved in hypersplenism in the condition of portal hypertension. CONCLUSION: cDNA microarray-based screening of differentially expressed genes in the macrophages in the spleen may provide new insights into the pathogenesis of hypersplenism in patients with portal hypertension. PMID- 17121697 TI - [Construction of a recombinant adenovirus vector for interferon-gamma-inducible protein 10 and the adenovirus preparation]. AB - OBJECTIVE: To construct a recombinant adenovirus vector for expressing interferon gamma-inducible protein 10 (IP-10) by homogenous bacterial recombination. METHODS: IP-10 gene was cloned into the shuttle plasmid pAdTrack-CMV that contained the coding sequence of enhanced green fluorescent protein (EGFP). The shuttle plasmid was then transformed into E. coli BJ5183 with pAdEasy-1 vector by chemical transformation. The recombinant adenovirus vector pAd/IP-10 was identified by enzyme digestion with Pac I and the linearized plasmid was transfected into HEK293 cells. RESULTS: The positive clones were identified with enzyme digestion and polymerase chain reaction (PCR) and were further verified by DNA sequencing. The recombinant adenovirus of high titration was obtained after transfection and packaging in HEK293 cells. CONCLUSION: A recombinant adenovirus vector for expression of IP-10 has been constructed successfully and high-titer active adenovirus is obtained for functional study of IP-10 protein. PMID- 17121698 TI - [Standardization of rat stable orthotopic liver transplantation model and comparison of the effect of two liver graft perfusion methods]. AB - OBJECTIVE: To standardize the establishment of a stable rat model of orthotopic liver transplantation and surgical technique of two-cuff technique for improving the operation success rate, and compare the effect of graft perfusion via the portal vein and the abdominal aorta on the graft function. METHODS: Totally 100 cases of orthotopic liver transplantation was performed in rats under microscope according to modified Kamada's two cuff technique. The rats were divided into 2 groups with the donor liver perfused through the portal vein with 10 ml cold lactated Ringer's solution, and via the abdominal aorta with 20 ml cold lactated Ringer's solution, respectively. The postoperative function recovery and pathological changes of the liver grafts were evaluated by serum ALT detection and histopathological examination. The operation success rate, 3-month survival rate of the rats and the complications were observed. RESULTS: No significant differences was noted in the liver function, operation success rate and 3-month survival rate between the 2 groups, and histopathological examination also showed similar findings. The success rates of the two groups were 98% and 96%, with 3 month survival rate of 93.5% (29/31) and 93.3% (28/30) (P>0.05), respectively. CONCLUSION: The liver transplantation models with portal vein or abdominal aorta graft perfusion both serve well their respective purposes. Good microsurgical skills, standardized performance and shortened anhepatic period are the keys to improved stability and survival rate and reduced operative complications. PMID- 17121699 TI - [Effects of Sini decoction on vascular stenosis after iliac artery balloon injury and oxidative stress in rabbits]. AB - OBJECTIVE: To investigate the effect of Sini decoction (SND) in preventing vascular restenosis and protecting against oxidative stress after rabbit iliac artery balloon injury. METHODS: Twenty-four male New Zealand albino rabbits were equally randomized into control group, model group and SND group. Rabbits in the control group were fed with common forage, and those in the model and SND groups with high-fat diet. Two weeks later, the iliac arteries of the rabbits in the latter two groups were subjected to balloon injury. Four weeks after the operation, the rabbits were killed and the vascular structure was observed by scanning electron microscope and optical microscope, with the serum cholesterol level, superoxide dismutase (SOD) activity and malondialdehyde (MDA) level determined. RESULTS: Scanning electron microscopy showed that the endothelial cell lining in the iliac artery of the control and SND group remain regular, but the arteries in the model group presented with desquamated and exposure of the collagen fibril beneath the endothelium. Optical microscope revealed narrowed vascular lumen, thicken intima and numerous arteriosclerotic plaques in the model group in comparison with the control group, whereas the vascular lumen and intima thickness remained basically normal in SND group. The levels of total cholesterol, low-density lipoprotein cholesterol and triglyceride were decreased with high-density lipoprotein cholesterol levels increased in SND group, which was not observed in the model group. The serum SOD activity was higher in the control group than in the model and SND groups, but SND group had higher serum SOD activity than the model group. The serum MDA level was lower in the control group than in the other two groups, but SND group had lower MDA level than the model group. CONCLUSION: SND can alleviate intimal hyperplasia and vascular stenosis in injured rabbit iliac artery, possibly in relation to increased SOD activity and decreased lipid peroxidation as a result of SND treatment. PMID- 17121700 TI - [Characteristics of uterine contraction and stages of labor under continuous epidural block anesthesia]. AB - OBJECTIVE: To observe the characteristics of uterine contraction and stages of labor during delivery under continuous epidural block anesthesia. METHODS: Totaling 213 parturients in spontaneous labor under epidural block anesthesia with dilated cervical orifice of 3 cm were monitored for the contraction cycle, duration, intensity and curve types of uterine contraction, and recordings were made for 30 min before and 30, 60 and 120 min after the anesthesia took effect, respectively. The duration of the active phase in the first, second and third stages of labor was compared between 421 cases with anesthesia and 237 without anesthesia. RESULTS: Significant difference was noted in the objective indexes of uterine contraction recorded after anesthesia had taken effect (P<0.05) in comparison with those before anesthesia, suggesting significantly attenuated uterine contraction after anesthesia, whereas these indexes underwent no significant further variation as compared between different time points after anesthesia (P>0.05). The average active phase in the first stage was significantly shorter in anesthesia group than that in the control group (P<0.05), but the average duration of the second and third stages of labor differed little between the two groups with appropriate use of oxytocin under strict monitoring (P>0.05). The rates of obstetric forceps utilization and use of oxytocin were higher in anesthesia group than in the control group (P<0.05). CONCLUSION: Epidural block anesthesia produces certain influences on uterine contraction and stages of labor during delivery, for which appropriate treatment measures may prove beneficial. PMID- 17121701 TI - [Spontaneous discharge modulation by acetylcholine in chronically compressed rat dorsal root ganglion neurons]. AB - OBJECTIVE: To study the effects of acetylcholine (ACh) on spontaneous discharges of compressed rat dorsal root ganglion (DRG) neurons. METHODS: In chronically compressed DRG model, the spontaneous discharge of a single fiber from the DRG neuron in response to ACh treatment was recorded. RESULTS: Active spontaneous discharges were recorded in the injured DRG, and 77.9% of the injured DRG neurons responded to Ach treatment in the manner of simple excitation, or excitation followed by inhibition. The responses were enhanced with the increase of Ach concentration. CONCLUSION: Injured DRG remains active in spontaneous discharges, which can be significantly influenced by ACh treatment. PMID- 17121702 TI - [Acquisition of fetal cells from transcervcial cells in early pregnancy and immunocytochemical study]. AB - OBJECTIVE: To study the presence of trophoblast cells in the lower uterus in early pregnancy, identify fetal cells by immunocytochemistry, and determine the best timing for sample collection. METHODS: Samples from healthy gravida in early pregnancy were divided into HE group (I) and immunocytochemistry group (II), and those from healthy nonpregnant women were also divided into HE group (III) and immunocytochemistry group (IV). Four different methods (usage of a cotton swab, aspiration of the cervical mucus, lavage of the endocervical canal, and lavage of the intrauterine cavity) were employed for collecting the samples, which were tested with HE staining or immunocytochemistry, and the presence of fetal cells was observed under optical microscope. RESULTS AND CONCLUSION: Fetal cells were detected in the genital tract of the gravida in early pregnancy. Utilization of the lavage of the endocervical canal or the lavage of the intrauterine cavity allowed greater amount of fetal cell acquisition, and sampling of the fetal cells between 50 days and 79 days of gestational age yielded optimal results. These sampling methods may provide safe and effective means for prenatal diagnosis with minimal invasiveness. PMID- 17121703 TI - [Establishment of miniCTTA multiplex set with fluorescence-labeled primers]. AB - OBJECTIVE: To establish a miniCTTA multiplex set including short tandem repeat (STR) markers as CSF1PO, TH01, and TPOX and amelogenin gene, whose amplified fragments are shorter than 130 bp. METHODS: The length of the gene fragments amplified with fluorescence-labeled primers were analyzed by ABI3100 Genetic Analyzer for genotyping of 100 unrelated individuals, 10 genealogies and 30 highly degraded specimens. RESULTS: The genotypes derived form miniCTTA were consistent with the results by AmpFLSTR Identifiler kit. CONCLUSION: MiniCTTA multiplex set is useful for personal identification and paternity test, especially applicable for degraded DNA sample. PMID- 17121704 TI - [Construction of artificial nerve bridge by three-dimensional culture of interleukin-1beta- activated Schwann cells with human hair keratins]. AB - OBJECTIVE: To culture interleukin-1beta (IL-1beta)-activated Schwann cells (SCs) with human hair keratins (HHKs) for artificial nerve bridge construction. METHODS: SCs purified by primary culture with or without IL-1beta activation were cultured with HHKs decorated by extracellular matrix (ECM), and the artificial nerve bridge was implanted into the defect of rat sciatic nerve. The morphology of the SCs cultured with HHKs was monitored by inverted microscope, scanning electron microscope and evaluated by immunocytochemical staining, and the expression of nerve growth factor (NGF) in the sciatic nerve was observed by in situ hybridization. RESULTS: Activated SCs showed better ability to adhere to the HHKs and grew well. The HHKs component in the artificial nerve bridge underwent degradation in the sciatic nerve defect after 3 to 4 weeks, and IL-1beta activation resulted in enhanced NGF expression in the SCs. CONCLUSION: The constructed artificial nerve bridge by three-dimensional culture of IL-1beta activiated SCs with HHKs decorated by ECM promotes the repair of sciatic nerve defects and accelerates sciatic nerve regeneration. PMID- 17121705 TI - [Effects of perindopril on left ventricular remodeling and myocardial osteopontin expression in rats with myocardial infarction]. AB - OBJECTIVE: To observe the effects of perindopril on left ventricular remodeling and myocardial osteopontin expression in rats with myocardial infarction (MI). METHODS: Male adult SD rats were randomly divided into sham-operation group, MI saline group and MI-perindopril group, and in the latter two groups, ligation of the left anterior descending artery was performed to establish rat models of myocardial infarction and perindopril (2 mg/kg daily) or saline was administered since the next day of MI. Four weeks later, the left ventricular diameter (LVEDD and LVESD) and left ventricular ejection fraction were estimated with echocardiography, and the left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP) and -/+dp/dt(max) was obtained via hemodynamic measurement, with also evaluation of the cardiac myocyte diameter and interstitial fibrosis infiltration with histological methods. Western blotting was performed to detect the expression level of myocardium osteopontin protein. RESULTS: Compared with the sham-operation group, all MI rats developed significant systolic and diastolic dysfunction, as indicated by decreased LVEF, LVSP and -/+dp/dt(max), as well as increased LVEDP. MI rats showed significantly dilated left ventricles and higher ventricular weight/body weight ratio, significantly increased cardiomyocyte diameter and marked interstitial fibrosis in the non-infarction area. Perindopril treatment partially prevented cardiac dysfunction and left ventricular remodeling as indicated by the above indices. No osteopontin was detected in the myocardium of sham-operation rats, and in MI rats, high level of osteopontin expression, as detected in MI-saline group, was significantly inhibited by perindopril treatment. CONCLUSION: Perindopril treatment can significantly inhibit left ventricular remodeling and myocardium osteopontin expression in rats with MI. PMID- 17121706 TI - [Expression of chemokine monokine induced by interferon-gama in patients with chronic hepatitis B]. AB - OBJECTIVE: To study the expression levels of monokine induced by interferon-gama; (Mig) mRNA and its association with HBV DNA and alanine aminotransferase (ALT) in patients with chronic hepatitis B. METHODS: The level of Mig mRNA in peripheral blood mononuclear cells (PBMCs) was dynamically detected with real-time quantitative PCR, and the ratio of chemokine/GAPDH was considered to represent the final chemokine level. The plasma level of was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The mean level of Mig mRNA in PBMCs of the patients with chronic hepatitis B was 0.6883-/+0.0693, which was significantly higher than that in normal controls (P<0.001). The plasma Mig level in the patients was 609.6-/+73.8 pg/ml, also significantly higher than that in normal controls (P<0.05). In patients with chronic hepatitis B, the level of Mig mRNA in the PBMCs was significantly correlated with plasma Mig level (r=0.7157, P<0.001), and plasma Mig level was correlated with plasma ALT level (r=0.7220, P<0.001) and plasma HBV DNA level (r=0.7266, P<0.001). CONCLUSION: Both the expression of Mig mRNA in PBMCs and plasma Mig concentration are elevated in patients with chronic hepatitis B. Mig plays an important role in migration of the inflammatory cells to the liver and mediates the development of chronic hepatitis B. PMID- 17121707 TI - [Nuclear ribosomal DNA internal transcribed spacer 1 sequences of 4 Leonurus species]. AB - OBJECTIVE: To analyze the nuclear ribosome DNA (nrDNA) internal transcribed spacer (ITS) sequences of 4 Leonurus species, and the possibility of using them for molecular authentication of the crude drugs from the genus. METHODS: The nrDNA ITS sequence (including ITS1, 5.8S rDNA, ITS2, and partial 18S rDNA and 26S rDNA) of L. japonicus and its 3 adulterant species were amplified and sequenced, and CLUSTRAL X and MEGA software was employed for analysis. RESULTS: The variation of ITS1 and ITS2 between L. japonicus and its adulterant species ranged between 7.2% and 18.8% and between 14.2% and 27%, respectively. The phylogenic tree derived from the dendrograms based on the ITS sequence data contained some discrepancy from the traditional classification. CONCLUSION: The nrDNA ITS sequences can be used potentially as efficient markers for identification of L. japonicus and its adulterants, and further study is needed for studying the phylogeny of Leonurus. PMID- 17121708 TI - [Auditory brainstem response in normal guinea pigs and those with gentamicin induced hearing loss under awake and anesthetic conditions]. AB - OBJECTIVE: To observe the alterations of auditory brainstem response (ABR) in guinea pigs with gentamicin-induced hearing loss under awake and anesthetic conditions. METHODS: We recorded the ABR in 20 normal guinea pigs and 20 with gentamicin-induced hearing loss before and after anesthesia for statistical analysis. RESULTS: No significant difference was observed in the waveform, response threshold (RT), I and III peak latencies (PL), I-III interpeak latencies (IPL) of ABR between awake and anesthetic conditions in normal guinea pigs (P>0.05), nor did gentamicin-induced hearing loss showed obvious impact on the ABR parameters (P>0.05). CONCLUSION: No significant ABR alterations occur under awake and anesthetic conditions in either normal guinea pigs or those with hearing loss, therefore ABR test can be performed without anesthesia to ensure the success and error minimization of the experiment. PMID- 17121709 TI - [Chemopreventive effect of celecoxib against DMBA-induced breast cancer and its mechanism]. AB - OBJECTIVE: To evaluate the chemopreventive effect of celecoxib, a specific cyclooxegenease-2 (COX-2) inhibitor, on chemically induced breast cancer of rats and its effect on COX-2 expression. METHODS: 7, 12-dimethylbenz anthracene (DMBA) was administered intragastrically in SD female rats to establish breast cancer models, which were divided subsequently into control group, tamoxifen group and celecoxib group to receive different treatments accordingly. The occurrence rate of breast cancer was observed and the effect of celecoxib on COX-2 and vascular endothelial growth factor (VEGF) expressions assayed by immunohistochemical SP method. RESULTS: The incidence of breast cancer in tamoxifen group (48.15%) and celecoxib group (50.00%) were both significantly lower than that in the control group (85.71%; P=0.003 and P=0.004, respectively). The positivity rate of COX-2 expression in celecoxib group (28.57%) was significantly lower than those of tamoxifen group (48.15%) and control group (83.33%; P=0.001 and P=0.035, respectively). The positivity rate of VEGF expression in celecoxib group (42.86%) was significantly lower than that of control group (79.17%, P=0.023), but comparable with that in tamoxifen group (46.15%, P=0.863). CONCLUSION: Celecoxib can significantly suppress DMBA-induced breast cancer in female rats possibly through down-regulation of COX-2 and VEGF expressions. PMID- 17121710 TI - [Anatomical study of venous spaces in transsphenoidal approach for cavernous sinus surgery]. AB - OBJECTIVE: To study the anatomical and morphological characteristics of the venous spaces involved in surgery via transsphenoidal approach to the cavernous sinus (CS). METHODS: Ten fixed cadaver heads (six male, four female) with red and blue latex injected in the arteries and veins, respectively, were used to perform the transsphenoidal approach. The anterior wall of the sphenoidal sinus and the floor of sellar turcica were opened as much as possible to expose the dura mater at the sellar floor and the inferior wall of CS, and the location of the anterior and inferior intercavernous sinuses were observed carefully. All the spaces of CS were observed and measured. According to the observations, the venous spaces available for operation were identified and analyzed. RESULTS: In all the cadaver heads, 4 anterior and 5 inferior intercavernous sinuses were found, with the former locating below the optic protuberance, while the latter situated at the turn of the sellar protuberance at the clival indentation. CS was subdivided into medial space, inferolateral space, and dorsolateral space. CONCLUSIONS: In transsphenoidal approach, opening of anterior and inferior intercavernous sinus is liable to result in intra- and postoperative venous bleeding, and understanding of the location of the intercavernous sinus and appropriate utilization of these CS may help reduce intraoperative vascular and nerve injury. PMID- 17121711 TI - [Evaluation of perioperative blood loss following total knee arthroplasty]. AB - OBJECTIVE: To evaluate perioperative occult blood loss following total knee arthroplasty (TKA). METHODS: A retrospective analysis of 40 patients undergoing TKA was conducted to calculate the mean blood loss and occult blood loss according to Gross formula. RESULTS: The mean total blood loss was 1538 ml in these cases with occult blood loss of 791 ml. In patients with autologous blood transfusion, the mean total blood loss was 1650 ml with occult blood loss of 786 ml. In patients without autologous blood transfusion, the mean total blood loss was 1370 ml with occult loss of 798 ml. CONCLUSION: TKA often results in large volume of occult blood loss in the perioperative period which can not be fully compensated by autologous blood transfusion, and additional blood supply is needed for maintenance of the circulating volume. PMID- 17121712 TI - [Cloning and expression of murine Toll-like receptor-2 N terminal and preparation of its antibody]. AB - OBJECTIVE: To prepare the recombinant murine Toll-like receptor-2 N-terminal (mTLR-2N) fusion protein and obtain anti-mTLR-2N polyclonal antibody. METHODS: The gene encoding 153 amino acids of mTLR-2N was amplified by PCR and cloned into pET32A vector with sequence verification. The recombinant fusion protein was expressed in E. coli and purified by Probond resin column. Rabbits were immunized with fusion protein to obtain the polyclonal anti-sera, and the antibodies were identified by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry. RESULTS: The recombinant fusion protein was efficiently expressed and purified. The polyclonal antibodies could bind to the fusion protein expressed in different vectors as the antigens in ELISA, and also bind with RAW264.7 cells expressing mTLR-2 and CHO cells transfected with full-length mTLR-2 gene. CONCLUSION: The recombinant mTLR-2N fusion protein is obtained and the anti-mTLR-2N polyclonal antibody can recognize natural mTLR-2 on the cell surface. PMID- 17121713 TI - [Expressions of RhoC and osteopontin in esophageal squamous carcinoma and association with the patients' prognosis]. AB - OBJECTIVE: To investigate the expressions of RhoC and osteopontin (OPN) protein in esophageal squamous carcinoma (ESC) and their association with the biological behavior of ESC. METHODS: The expressions of RhoC and OPN protein were detected in 80 ESC cases by immunohistochemistry. RESULTS: The positive expression rate of RhoC was 66.25% in these ESC cases. The rate was significantly higher in cases with lymph node metastasis than in those without (r(s)=-2.115, P<0.05), but RhoC expression was not associated with the tumor diameter, differentiation or TNM grade (P>0.05). The RhoC-positive patients had significantly shorter survival time than the negative patients (P<0.001). All the 80 ESC patients were positive for OPN expression, and OPN expression levels were correlated with the differentiation (chi(2)=10.766, P<0.05) and lymph node metastasis of the tumor (r(s)=-2.289, P<0.05), but not with the tumor diameter or TNM grade (P>0.05). Higher expression level of OPN was closely related to shorter survival time of the patients (P<0.05). A positive correlation was found between RhoC protein and OPN expressions (r(s)= 0.408, P<0.001) in these cases. CONCLUSION: The expressions of RhoC and OPN protein are closely related to lymph node metastasis of ESC and the patients survival time, and therefore may serve the purpose of prognostic evaluation of ESC. PMID- 17121714 TI - [Simulation and experimental study of cone artifact in spiral CT]. AB - OBJECTIVE: To study regular patterns of cone artifact resulted from interpolation algorithm of spiral CT. METHODS: Based on the principle of interpolation algorithm and back-projection reconstruction, a mathematical model of the reconstructed image was established to clarify the relation of the scanning parameters and the characteristics of the scanned object with the cone artifact. Experiments were carried out by a set of acrylic phantoms on siemens plus 4 CT scanners. RESULTS: The artifact in the image was directly proportional to the table increment per gantry rotation of the scanner, and was positively correlated to the tangent of half cone-angle and inversely to the radii in the reconstruction plane of the phantom. The theoretical analysis was validated by experimental results. CONCLUSION: The cone artifact is related to the scanning parameters and the characteristics of the scanned object. PMID- 17121715 TI - [Isolation and in vitro culture of follicular papilla cells from rat vibrissae]. AB - OBJECTIVE: To establish a simple method for isolating and culturing follicular papilla cells from rat vibrissae. METHODS: The intact follicles were obtained and digested in 0.2% collagenase I with agitation on a rotary stirrer at 100 r/min at 37 degrees C; for 3 h. The suspension was centrifuged at 300 r/min and the papilla cells were collected and suspended in DMEM for cell culture. The adhesion efficiency of the dermal papilla cells was evaluated and compared with that of the cells obtained by microdissection. RESULTS AND CONCLUSION: The described procedure allowed efficient and rapid isolation of the dermal papilla cells from rat vibrissae and ensured improved adhesion of the dermal papillae and outgrowth of the cells with reduced labor and risk of contamination. The cells obtained with this procedure were positive for alpha-smooth muscle actin staining. PMID- 17121716 TI - [Radioprotective effect of catechines against radiation injury in mice]. AB - OBJECTIVE: To study the radioprotective effect of catechines against radiation injury in mice. METHODS: Catechines were administered in mice intragastrically at the daily dose of 200 mg/kg for 10 consecutive days before whole body irradiation with 6 Gy X-rays. The body weight changes, survival time, 30-day survival rate, and counts of peripheral white blood cells were recorded. RESULTS: The mice with catechine pre-treatment before X-ray exposure suffered less body weight loss than those without the treatment before exposure. Catechines markedly increased the survival time of the irradiated mice, and raised the 30-d survival rate of the irradiated mice to 53.33% as compared with the rate of 13.33% in the radiated mice without catechine pre-treatment. Catechines significantly promoted recovery of peripheral white blood cells. CONCLUSION: Catechines have definite radioprotective effect against radiation injury in mice. PMID- 17121717 TI - [Cytotoxic effect of oncolytic virus combined with mitomycin against human bladder cancer cells in vitro and in vivo]. AB - OBJECTIVE: To evaluate the effect of combined use of oncolytic virus and the chemotherapeutic agents mitomycin (MMC) in growth inhibition of human bladder cancer cell line T-24 in vitro. METHODS: Human bladder cancer cell line T-24 was infected with oncolytic virus (ONYX-015) of different multiplicity of infection, or treated with MMC in addition to ONYX-015. The changes in the cell growth, morphology, and apoptosis of cultured T-24 cells were observed by means of cell counting and fluorescence microscopy after the treatments. The effects of the treatment protocols were also tested in nude mouse model of implanted subcutaneous tumor. RESULTS: Combined use of ONYX-015 and MMC produced substantially stronger cytotoxic effect against T-24 cells than exclusive use of ONYX-015. In in vivo experiments, combination of oncolytic virus and MMC resulted in much more significant tumor growth inhibition than either of the agents used alone. Obvious T-24 cell apoptosis could be observed in response to combined ONYX 105 and MMC treatment and exclusive ONYX-105 treatment. CONCLUSIONS: ONYX-015 combined with MMC can produce significant cytotoxicity against T-24 cells and enhance therapeutic efficacy against bladder carcinoma. PMID- 17121718 TI - [Percutaneous radiofrequency ablation combined with transcatheter arterial chemoembolization and percutaneous ethanol injection for recurrent small hepatocellular carcinoma]. AB - OBJECTIVE: To explore the value of percutaneous radiofrequency ablation (PRFA) combined with transcatheter arterial chemoembolization (TACE) and percutaneous ethanol injection (PEI) in the management of recurrent small hepatocellular carcinoma. METHODS: Between March 2001 and March 2005, 52 patients with recurrent hepatocellular carcinoma (tumor size< or =5 cm) underwent PRFA, and 14 of the patients (tumor size 3-5 cm) also received TACE and PEI, and their clinical data were analyzed retrospectively. RESULTS: MRI or CT after PRFA revealed complete coagulative necrosis of the tumor in 38 cases (tumor size <3 cm). In the 14 patients (tumor size 3-5 cm) with also TACE and PEI, complete necrosis occurred in 11 cases (78.6%). In the patients involved in this study, the 1-, 2-, 3- and 4 year survival rates were 96.2%, 69.4%, 45.5% and 30.0%, respectively. CONCLUSIONS: PRFA is an effective modality for local treatment of recurrent small hepatocellular carcinoma, capable of total elimination of tumors <3 cm. For tumors of 3-5 cm, combination with TACE and PEI may help increase the tumor necrosis rate following the ablation and raise the patients' survival rate. PMID- 17121719 TI - [Isolation, in vitro culture and identification of cardiac stem cells from neonatal SD rats]. AB - OBJECTIVE: To isolate and culture cardiac stem cells (CSCs) in vitro and evaluate their potential of differentiation into functional cardiac myocytes. METHODS: Myocardial tissues obtained from neonatal SD rats were cut into pieces of 0.5-1.0 mm(3), and digested twice for 5 min at 37 degrees C; with 0.2% trypsin and 0.1% collagenase II. The remaining tissues were cultured in complete explant culture medium (CEM) at 37 degrees C; in the presence of 5% CO(2). About a week later, a layer of fibroblast-like cells was generated from the adherent explants. These cells were passaged and seeded at about 1x10(6) cells/ml in poly-D-lysine-coated multi-well plates in cardiosphere-growing medium. When beating of the cultured cells was observed (at week 2), flow cytometry and immunohistochemistry were performed for identification of the primary and passaged cells. RESULTS: The primary cells were successfully cultured from the digested myocardial tissue, and flow cytometry demonstrated the phenotype of c-kit(+)CD31(+)CD34(-)CD45(-)CTnT( ). After cell passage for about two weeks, single beating cells and cell clusters with synchronized contraction were seen microscopically, and their phenotype was converted to c-kit(+)CD31(-)CD34(-)CD45(-) CTnT(+). Immunohistochemistry staining identified CTnT expression in the passaged cells but not in the primary cells. CONCLUSIONS: A cell population with the phenotype c-kit(+)CD31(+)CD34(-)CD45( )CTnT(-) has been obtained from neonatal SD rat heart, which possesses the potential to differentiate in vitro into beating cardiac myocytes and express CTnT protein. PMID- 17121720 TI - [Effect of calcineurin Aalpha gene overexpression on the myocardium apoptosis induced by hypoxia-reoxygenation and adrenergic receptors]. AB - OBJECTIVE: To investigate the effect of calcineurin AalphacDNA (AdCnAalpha) overexpression as a result of adenovirally mediated gene transfer on neonatal rat cardiac myocyte apoptosis induced by hypoxia-reoxygenation (H/R) and adrenergic receptors. METHODS: Neonatal rat cardiac myocytes were cultured for 20 h after AdCnAalpha transfection, and treated with isoproterenol (10 micromol/L) and 24 h of hypoxia followed by 4 h of reoxygenation (24H/4R). The cardiac myocyte apoptosis induced by the treatments was assessed by flow cytometry and DNA laddering, and the levels of calcineurin, p38 and phosphorylation p38 (p-p38) were determined by Western blotting and (or) RT-PCR. RESULTS: AdCnAalpha transfection promoted cultured neonatal rat cardiac myocyte apoptosis induced by isoproterenol+24H/4R as compared with the treated cells without transfection (14.247-/+0.525 vs 10.763-/+1.554, P<0.01), along with greater phosphorylation p38 protein expression (1.60-/+0.22 vs 2.42-/+0.19, P<0.01). The levels of p38 underwent no obvious change after AdCnAalpha transfection (P<0.05). CONCLUSIONS: AdCnAalpha transfection can promote cardiac myocyte apoptosis induced by H/R and adrenergic receptors, the mechanism of which might be associated with p38 mitongen-activated protein kinase (p38MAPK) activation. PMID- 17121721 TI - [Dynamic changes of plasma VEGF, SDF-1 and peripheral CD34+ cells in patients with acute myocardial infarction]. AB - OBJECTIVE: To investigate the changes in plasma levels of vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1 (SDF-1) and in peripheral CD34(+) cells in patients with acute myocardial infarction (AMI), and explore their role in AMI. METHODS: Enzyme-linked immunoassay (ELISA) was employed for measuring the levels of VEGF and SDF-1 in AMI patients on days 1, 3, 7, 10, and 14 of onset and in normal control subjects. The absolute counts of CD34(+) in the peripheral blood were measured on days 1, 7, and 14 by flow cytometry in AMI patients, with their myocardial enzyme and troponin I detected and electrocardiography (ECG) and echocardiography (UCG) recorded. RESULTS: Peripheral CD34(+) cells obviously increased on day 7 after AMI onset (2.35 /+0.72/microl vs 1.48-/+0.49/micro, P<0.05). VEGF levels were significantly higher in AMI patients than in the control subjects, reaching the peak level and on day 14 (197.56-/+39.87 vs 53.79-/+18.12 pg/ml, P<0.01). SDF-1 level obviously decreased on day 1 after AMI onset (1683.12-/+224.79 vs 2178.67-/+265.34 pg/ml, P<0.01), followed by gradually increased to the control level. Obvious correlation was noted between the level of VEGF on day 7 and the peak level of peripheral CD34(+) cells, and the peak plasma VEGF level was obviously associated with the peak serum CK-MB and troponin I levels. CONCLUSION: The stem cells are mobilized into the peripheral blood in the event of AMI. Obviously increased VEGF level following AMI may persist for at least 2 weeks, whereas SDF-1 level undergoes temporary decrement after AMI. The dynamic changes of VEGF and SDF-1 can be related to the mobilization and homing of the stem cells to the injured myocardium. PMID- 17121722 TI - [Construction and identification of recombinant adenoviruses containing the gene fragments encoding Her2/neu extracellular and transmembrane domains]. AB - OBJECTIVE: To construct eukaryotic expression vectors using recombinant adenovirus containing the gene fragments encoding Her2/neu extracellular first ligand-binding domain (Her2-ECD), full-length extracellular domain (Her2-ECD), and extracellular and transmembrane domain (Her2-TM). METHODS: The cDNAs were amplified by RT-PCR and inserted into shuttle pAdTrack-CMV plasmids. Viral plasmids were obtained from homologous recombination in E. coli BJ5183, and transfected into 293 cells via liposome. Formation of viral plaque and expression of green fluorescent protein were observed by fluorescence microscopy, and the target proteins were detected by Western blotting. RESULTS: The target cDNA fragments were amplified by PCR with expected lengths and the DNA sequences were confirmed against Genbank. Formation of viral plaque, expression of green fluorescent protein and the target proteins were detected in 293 cells transfected by the viral plasmids, which showed elevated expression of Her2/neu protein with the increase of multiplicity of infection (MOI). CONCLUSION: The eukaryotic expression vectors using recombinant adenovirus have been successfully constructed for expression of Her2/neu extracellular and transmembrane domains. PMID- 17121723 TI - [Antisense oligonucleotide against survivin induces apoptosis and enhances adriamycin sensitivity of SMMC-7721/ADM cells]. AB - OBJECTIVE: To explore the effects of survivin-specific antisense oligonucleotide (ASODN) delivered via liposome on the growth and apoptosis of drug-resistant human hepatic cancer cell line SMMC-7721/ADM and the sensitivity of the cells to adriamycin (ADM). METHODS: SMMC-7721/ADM cells were divided into 6 groups and treated with liposome, ADM, sense oligonucleotide (SODN), SODN+ADM, 400 ng/ml ASODN, and 400 ng/ml ASODN+ADM group, respectively. MTT assay was used to calculate the relative survival rates of the cells, and the changes in cell apoptosis and cycle were detected with flow cytometry. RT-PCR and Western blotting were performed to detect the expressions of survivin mRNA and protein, respectively. RESULTS: The apoptotic rate of ASODN-treated cells was much higher than that of the control cells. survivin protein expression showed no significant variation between cells treated with liposome, ADM, SODN, and SODN+ADM (P>0.05), whereas compared with these 4 groups, cells treated with 400 ng/ml ASODN and 400 ng/ml ASODN+ADM had significantly lowered survivin mRNA expression (P<0.05), without significant differences between the latter two groups (P>0.05). SMMC 7721/ADM cells cultured in the presence of ASODN and adriamycin showed significant growth inhibition in comparison with ASODN group and ADM group. CONCLUSION: survivin-specific ASODN can enhance the sensitivity of SMMC-7721/ADM cells to ADM by depressing survivin expression in the cells, thus improves the effect of ADM chemotherapy for liver cancer. PMID- 17121724 TI - [Diffusion tensor imaging of the white matter tracts in preoperative patients with cerebral neoplasm]. AB - OBJECTIVE: To evaluate the quantitative relationship between white matter tract (WMT) variation resulting from cerebral tumors and shifting of the fractional anisotropy (FA) index in magnetic resonance (MR) diffusion tensor imaging (DTI). METHODS: Four female and 8 male patients aged from 21 to 62 years with brain malignancies (2 malignant lymphomas, 2 low-grade astrocytomas, and 8 high-grade cerebral gliomas) underwent conventional contrast-enhanced MR and DTI examinations before operation. Routine T(2)-weighted image, fractional anisotropic (FA) map, color-coded directional map, three-dimensional white matter tractography (WMT), and the FA index of bilateral internal capsule were obtained in every patient. RESULTS: Fiber tractography derived from DTI was consistent with known white matter fiber anatomy. The DTI patterns in WMT altered by the tumor were categorized on the basis of FA1/FA2 ratio as follows: pattern 1, FA1/FA2> or =75% with normal or only slightly decreased FA; pattern 2, 50%< or =FA1/FA2<75% with WMT displacement; pattern 3, 25%< or =FA1/FA2/50% with WMT involvement; pattern 4, FA1/FA2<25% with WMT destruction. CONCLUSIONS: DTI allows for visualization of WMT and benefits surgical planning for patients with intrinsic brain tumor. There is a positive relationship between the bilateral FA ratio (FA1/FA2) variation and WMT alterations resulting from the tumor. PMID- 17121725 TI - [Serum sCD40L detection for risk evaluation of acute coronary syndromes]. AB - OBJECTIVE: To investigate the value of serum soluble CD40 ligand (sCD40L) detection in risk evaluation of acute coronary syndromes (ACS). METHODS: This study involved 200 patients with established diagnosis of ACS, with death or nonfatal myocardial infarction as the end point of observation during the 6-month long follow-up. Blood samples were obtained from the patients within the initial 72 h of ACS onset, and the levels of sCD40L and C-reactive protein (CRP) were determined with enzyme-linked immunosorbent assay (ELISA). Cardiac troponin I (cTnI) measurement was performed using chemiluminescent immunoassay. RESULTS: Of the 200 patients, 108 had serum sCD40L levels higher than 5.0 microg/L, and the levels of sCD40L, CRP and cTnI were found to significantly correlate with ACS. CONCLUSION: Independent detection of serum sCD40L, CRP and cTnI can help predict the risks of ACS, and their combined measurement may increase the sensitivity of the risk prediction and provide new cardiac makers to replace the cardiac enzymes for laboratory diagnosis and risk evaluation of cardiovascular events. PMID- 17121726 TI - [Effect of huperzine A on cerebral cholinesterase and acetylcholine in elderly patients during recovery from general anesthesia]. AB - OBJECTIVE: To observe the effect of huperzine A on cerebral cholinergic system in elderly patients during recovery from general anesthesia. METHODS: Thirty elderly patients undergoing elective surgery under general anesthesia were randomized in a double-blind manner into group I (n=15) to receive huperzine A (0.3 mg/2 ml) and group II (n=15) with normal saline (2 ml) given intravenously. Huperzine A or normal saline was administered 30 min before completion of the operation, and acetylcholine (Ach) concentration in the cerebral spinal fluid (CSF) of the patients was determined using high-performance liquid chromatography with electrochemical detector (HPLC-ECD) and the activity of cholinesterase inhibitors (ChE) evaluated with automatic biochemistry analyzer before general anesthesia induction (T1) and 5 h after operation completion (T2). RESULTS: In both the groups, Ach concentration in the CSF were lower at T2 than that at T1 (P<0.01), and at T2, CSF Ach concentration was significantly higher in group I than in group II (P<0.01); in group I, the activity of CSF ChE at T2 was lower than that at T1 (P<0.01), and also lower than at T2 in group II (P<0.01). CONCLUSION: Huperzine A can inhibit cholinesterase to increase Ach, which has a positive effect on cerebral cholinergic system in elderly patients during recovery from general anesthesia. PMID- 17121727 TI - [Low-dose ketamine combined with fentanyl for intravenous postoperative analgesia in elderly patients]. AB - OBJECTIVE: To investigate the clinical efficacy of and complications arising from low-dose ketamine combined with fentanyl for intravenous postoperative analgesia in comparison with the exclusive use of fentanyl in elderly patients. METHODS: Eighty elderly patients were randomized into two equal groups following thoracoabdominal surgery, and received intravenous analgesia with the combination of 0.5 mg/ml ketamine, 5.0 microg/ml fentanyl and 50 microg/ml midazolam (KF group) and with 7.5 microg/ml fentanyl plus 50 microg/ml midazolam (FT group), respectively. The drugs used were diluted in 200 ml normal saline. For analgesic administration, a loading dose (2-4 ml) was given followed by a background infusion (2.5-3.5 ml), with patient-controlled bolus doses of 2.0-3.0 ml with lock-out time of 20 min via PCA pump (Automedical, Korea). The static pain score (VAS), sedation score, and incidences of nausea, vomiting, pruritus and hallucinations were recorded during the initial 48 h after the surgery. RESULTS: The total analgesic dosage and PCA dosage in the two groups were similar (P>0.05). With similar VAS in the two groups (P>0.05), the sedative effect in KF group was much better than that in FT group (P<0.05) during the initial 48 h postoperatively. The incidences of nausea, vomiting and itching were lower in KF group than in FT group (P<0.05), and no illusion was reported in two groups during the initial 48 h. CONCLUSION: For producing comparable postoperative analgesic effect, low-dose ketamine combined with fentanyl can markedly reduce fentanyl requirement in the elderly patients and lowers the incidences of nausea, vomiting and itching in comparison with the exclusive use of fentanyl. PMID- 17121728 TI - [Relation of plasma brain natriuretic peptide to serum creatine kinase MB level in patients with acute myocardial infarction following primary percutaneous coronary intervention]. AB - OBJECTIVE: To study the relation between plasma brain natriuretic peptide (BNP) and serum creatine kinase MB (CK-MB) level in patients with acute myocardial infarction (AMI) following primary percutaneous coronary intervention (PCI). METHODS: Sixty-three consecutive patients with AMI were divided into two groups according to the timing of PCI, namely direct PCI and indirect PCI groups. Plasma BNP levels were measured in all patients on admission and at 4, 24 and 48 h after admission. The CK-MB level was measured every 3 h on the first day of hospitalization, every 6 h on the second day and every 12 h on the third day. RESULTS: BNP level increased gradually following admission and began to decrease 48 h after admission in the two groups of patients. The peak BNP level occurred at 24 h after admission, and the BNP levels in patients of indirect PCI group were significantly higher than that of direct PCI group at 4, 24 and 48 h after admission. The peak CK-MB level of the direct PCI group occurred significantly earlier than that of the indirect group. CONCLUSION: Plasma BNP level may serve as an important objective indicator for recanalization of the infarct-related arteries following PCI in the early stage of AMI, which can help in the decision on clinical treatment plans for AMI. PMID- 17121729 TI - The World Veterinary Poultry Association: the beginnings and first 25 years. AB - The World Veterinary Poultry Association (WVPA) was formed in 1959, the honour of being the first President of the Association going to Professor de Blieck and that of Secretary to Dr R.F. Gordon, who had both championed its formation. The First WVPA Conference (the name "Congress" was not applied until the third meeting) took place in Utrecht in 1960. The second Conference (1962) and the third Congress (1965) were in Cambridge and Paris, respectively. The fourth Congress, in Belgrade (1969), was a landmark one for two reasons: firstly, submitted papers were introduced, and secondly, the notion of a WVPA journal, later to be called Avian Pathology, was first discussed. A year later (1970) the journal came into being, Professor Klimes (Czhechoslovakia), the driving force behind its creation, being the first Editor-in-Chief, with Professor Kosic (Yugoslavia) as production Editor. Due to the ill health of Professor Klimes, Peter Biggs was confirmed as Editor-in-Chief in 1973. A charitable company, Avian Pathology Ltd, was formed in 1980, under which Avian Pathology was published. Dr L.N. (Jim) Payne succeeded Dr Biggs as Editor-in-Chief in 1988. The fifth and six Congresses were in Munich (1973) and Atlanta, Georgia, USA (1977), respectively. Four years later, at the Oslo Congress, it was agreed that the rather crude logo adopted at the fourth Congress should improved. This was done and is the current logo. On the 25th anniversary (1985) of the first WVPA Congress, the eighth Congress was held in Jerusalem. Membership had increased to almost 1000, with members in 39 countries, 22 of which had branches of WVPA. In 4 years it will be time to publish an account of the second 25 years in the history of the WVPA. PMID- 17121730 TI - Interaction between a live avian pneumovirus vaccine and two different Newcastle disease virus vaccines in broiler chickens with maternal antibodies to Newcastle disease virus. AB - Broiler chicks with maternal antibodies to Newcastle disease virus (NDV) but none to avian metapneumovirus (APV) were divided into six groups. One group was kept as an unvaccinated control group. Three of the other groups were vaccinated at 1 day old with live APV vaccine or one of two live NDV vaccines (VG/GA or HB1). The remaining two groups received the APV vaccine in combination with either of the two NDV vaccines at 1 day old. At intervals after vaccination for up to 42 days, distribution of the viruses in the tissues was monitored, together with humoral antibody responses. Few NDV isolations were made from any NDV-vaccinated chicks, probably due to the presence of NDV maternal antibodies. In both dual-vaccinated groups, APV persisted longer (up to 21 days post vaccination (d.p.v.)) than in the single vaccinates (up to 14 d.p.v.). After 14 d.p.v., antibody titres against APV in both dual-vaccinated groups remained higher than the single APV vaccinates. For NDV haemagglutination inhibition antibodies, similar titres were found in the single and dual NDV VG/GA vaccinates. However, for chickens dually vaccinated with NDV HB1 and APV, the haemagglutination inhibition titres were significantly higher at 21 and 28 d.p.v. than the single HB1 vaccinates. These differences reflect the fact that NDV haemagglutination inhibition titres may depend on the NDV vaccine used. PMID- 17121731 TI - Isolation and molecular characterization of a new Muscovy duck parvovirus from Muscovy ducks in the USA. AB - Between 1997 and 1999 several cases of a new disease in Muscovy ducks were reported in Pennsylvania, USA. The cases were characterized by locomotor dysfunction, weakness, recumbency, 40 to 60% morbidity and 10 to 40% mortality. The most characteristic microscopic lesions were moderate to severe degenerative rhabodomyopathy. In order to characterize the aetiological agent, virus isolation was attempted from the spleen, liver, heart, skeletal muscle and intestine by inoculation of 14-day-old Muscovy duck embryos with tissue homogenates. Deaths occurred on the second egg passage and parvoviruses were isolated by serial passage of allantoic fluid from dead embryos and then in Muscovy duck embryo fibroblast (MDEF) cultures. Parvovirus particles were observed in allantoic fluids and supernatants of MDEF cultures by transmission electron microscopy. Two genomic fragments, comprising 1108 nucleotides of the right open reading frame that codes for the structural viral proteins 1, 2 and 3, were amplified by polymerase chain reaction from one of the isolates, Muscovy duck parvovirus (MDPV)/PSU-31010. Comparison of this fragment with available sequences of other MDPV and related goose parvovirus (GPV) isolates showed that it had only 84.5% sequence identity with other MDPV isolates and 84.6% identity with the GPV isolates. This region shares over 99% identity among previously sequenced MDPV isolates and 95% identity among the related GPV isolates. This suggests that MDPV/PSU-31010 is divergent from all other sequenced MDPV and GPV isolates, and may represent a new group of avian parvoviruses. PMID- 17121732 TI - An outbreak of avian influenza subtype H9N8 among chickens in South Korea. AB - Low pathogenic avian influenza subtype H9N8 was diagnosed on a Korean native chicken farm in Gyeonggi province, South Korea, in late April 2004. Clinical signs included moderate respiratory distress, depression, mild diarrhoea, loss of appetite and a slightly elevated mortality (1.4% in 5 days). Pathologically, mucopurulent tracheitis and air sacculitis were prominently found with urate renal deposition. The isolated A/chicken/Kr/164/04 (H9N8) had an Ala-Ser-Gly-Arg (A/S/G/R) motif at the cleavage site of haemagglutinin, which has been commonly found in H9N2 isolated from Korean poultry. Phylogenetic analysis of the haemagglutinin and neuraminidase genes of the H9N8 avian influenza virus (AIV) isolate showed that reassortment had occurred. Its haemagglutinin gene was similar to that of Korean H9N2 AIVs, but its neuraminidase gene was closely related to that of A/WBF/Kr/KCA16/03 (H3N8) isolated from the faeces of wild birds in Korea. The pathogenicity of the isolate was tested on 6-week-old specific pathogen free chickens. The inoculated virus (H9N8) was recovered from most tested organs, including the trachea, lung, kidney, spleen, and caecal tonsil. This is the first report of an outbreak of low pathogenic avian influenza in chickens caused by AIV subtype H9N8. PMID- 17121733 TI - Relationship between mortality, clinical signs and tracheal pathology in infectious laryngotracheitis. AB - Previous studies in our laboratory using a combination of polymerase chain reaction and restriction fragment length polymorphism have identified at least five different genotypes of infectious laryngotracheitis virus (ILTV). However, the virulence of these classes of ILTV was not investigated. In this study, five groups (16 birds each) of 3-week-old specific pathogen free chickens were inoculated via the intratracheal route with 10(3) median embryo infected dose of five different strains of ILTV. Three further groups of chickens were inoculated similarly with the vaccine strains SA2 and A20 or with sterile phosphate-buffered saline (PBS) for comparison. Four days post-inoculation, clinical signs were monitored for scoring, and eight chickens from each group were subsequently euthanized, weighed and subjected to pathological and histopathological examinations. The remaining birds were monitored for clinical signs and mortality until 21 days post-inoculation. All groups inoculated with ILTV strains showed moderate to severe clinical signs 4 days after inoculation. The strain Q1-96 caused only minimal breathing symptoms with a median score that was not significantly different to that of the group inoculated with PBS, but was significantly different to those of the groups inoculated with other ILTV strains. The strain Q1-96 caused severe photophobia and conjunctivitis with a median score that was significantly higher than those of all other groups except for the group inoculated with the strain N3-04. All ILTV strains caused a significant reduction in weight gain when compared with the group inoculated with PBS. The strain Q1-96 caused an average weigh loss of 14% that was significantly higher than those of other ILTV strains. The strains S2-04 and Q1-96 induced only minor microscopic tracheal lesions while all the other ILTV strains, including the vaccine strains A20 and SA2, induced moderate to severe microscopic tracheal lesions. Median scores for microscopic tracheal lesions were well correlated with the number of viral genomes detected in trachea. The results revealed that there is considerable variation among ILTV strains in their tropism for trachea or conjunctiva. In addition it was revealed that ILTV strains with high affinity for conjunctiva can severely affect weigh gain. The ILTV numbers and microscopic lesions in trachea were not found to be reliable indicators of virulence since they are not necessarily correlated with mortality rate in ILT. PMID- 17121734 TI - Epidemiological and experimental evidence for immunodeficiency affecting avian infectious bronchitis. AB - We evaluated the effects of viral immunodeficiency on the outcome of infectious bronchitis virus (IBV) infection in chickens as a hypothetical cause for failure of adequate protection in vaccinated chickens. Initially, we investigated IBV isolations from cases of respiratory disease in association with the presence of thymic and/or bursal atrophy in 322 submissions during 1997 to 2002. Arkansas (Ark)-type IBV was most frequently isolated in spite of extensive ArkDPI vaccination in the broiler industry. The number of IBV isolations was consistently higher in broilers aged 27 to 43 days, coinciding with lymphocytic depletion of the bursa and/or thymus, providing circumstantial evidence that immunodeficiency and IBV incidence may be linked. S1 gene sequence analyses, antigenic characterizations, and challenge of susceptible chickens demonstrated that the field IBV isolates tested were closely related to vaccine strains and had low pathogenicity for chickens. We experimentally evaluated the effects of immunodeficiency caused by co-infection with chicken anaemia virus and infectious bursal disease virus on the outcome of IBV infection. Clinical signs and histological lesions were more persistent in immunodeficient chickens. Local specific IgA production was delayed and lower levels were achieved in immunodeficient chickens. At the same time, IBV RNA concentrations in tracheas and lachrymal fluids were higher and more persistent in immunodeficient chickens. Collectively, these results indicate that viral immunodeficiency most probably plays a relevant role in the epidemiology and outcome of IBV infection. PMID- 17121735 TI - The feathering gene is linked to degranulation and oxidative burst not cytokine/chemokine mRNA expression levels or Salmonella enteritidis organ invasion in broilers. AB - In the past, we showed differences in heterophil function between parental broilers (A [fast feathering] > B [slow feathering]) and their F1 reciprocal crosses (D [fast feathering] > C [slow feathering]). In the present study, we evaluated the linkage of the feathering gene to heterophil function, pro inflammatory cytokine/chemokine mRNA expression levels, and resistance to Salmonella enteritidis organ invasion. Heterophils were isolated from 2-day-old chickens (C and D) separated into males and females - slow males and females (SM and SF), and fast males and females (FM and FF). Heterophil functions of degranulation and oxidative burst were measured. Heterophils from FF chickens (183+/-8.9) released more (P < 0.05) beta-d-glucuronidase (microM) than heterophils from SF chickens (149+/-3.7); FF heterophils (4.6 x 10(4)) generated a significantly (P < 0.05) greater oxidative burst (mean relative fluorescent units) compared with SF heterophils (4.2 x 10(4)). Interleukin-6, CXCLi2, and interferon-alpha mRNA expression levels were quantitated by real-time quantitative reverse transcriptase-polymerase chain reaction. No differences were observed between SM and FM or between SF and FF heterophils. Finally, 1-day-old chickens were administered S. enteritidis and liver/spleen organ invasion was quantitated. No differences were observed between the number of S. enteritidis positive FF and SF chickens, but FM were significantly (P < 0.05) more resistant to S. enteritidis organ invasion than SM chickens. The data indicate degranulation and oxidative burst were linked with the feathering gene; however, interleukin-6, CXCLi2, and interferon-alpha mRNA expression levels were not. Furthermore, susceptibility to in vitro S. enteritidis organ invasion was not linked to the feathering gene. PMID- 17121736 TI - Viral load in 1-day-old and 6-week-old chickens infected with chicken anaemia virus by the intraocular route. AB - Although the effects of chicken anaemia virus (CAV) infection have frequently been investigated in young chickens, there have been few studies of the pathogenesis of CAV infection in older birds. The aim of the work reported here was to study viral loads in 6-week-old chickens and to compare these with those seen in younger birds. Specific pathogen free chickens were inoculated at 1 day or at 6 weeks of age with 10(4) median tissue culture infective doses of CAV by the intraocular route. Chicks infected when 1 day old were euthanized at day 14, 18 or 22 post inoculation (p.i.), and those infected when 6 weeks old at day 16, 18 or 20 p.i. Their body and thymus weights were determined and samples were collected from their spleen, liver and thymus. A quantitative polymerase chain reaction assay was developed and used to determine the number of viral genome copies in the tissue samples. In both age groups, viral genome concentrations increased in all organs up to day 18 p.i. and reached a peak in the spleen and liver at day 18 p.i. The peak viral concentrations in the thymus were detected at day 18 in the younger birds and at day 20 p.i. in older chickens. These studies have shown that exposure to CAV in older birds leads to similar levels of active viral replication to those seen in younger birds, and may result in subclinical infections in older birds with the potential to increase susceptibility to other infectious agents. PMID- 17121737 TI - Deposition of differently sized airborne microspheres in the respiratory tract of chickens. AB - As a part of the development of an efficient dry powder aerosol vaccine for poultry, the objective of this study was to accurately determine the deposition pattern of nebulized microspheres in the airways of unanaesthetized chickens of different ages (1 day, 2 weeks and 4 weeks old). In the first part of the study, the aerosol administration method was characterized: the influence of different nebulizers and nebulizing protocols on the relative humidity in the exposure chamber, the particle size distributions, the microsphere output and single microsphere percentage were determined. In the second part, birds were exposed to nebulized fluorescently labelled polystyrene microspheres (1 to 20 microm). Respiratory and gastro-intestinal tract tissue samples were collected and the number of fluorescent microspheres per sample was determined. In 2-week-old and 4 week-old chickens, microspheres of 5 and 10 microm, respectively, were too large for deposition in the lungs and air sacs as less than 5% of these microspheres penetrated into the lower airways. The larger size of microspheres reaching the lower airways of 4-week-old birds was explained by increasing airway dimensions with age. For 1-day-old chickens, deposition in the lungs decreased from 17 to 3% with increasing particle size (1 to 20 microm), but increased in the air sacs from 6 to 20%. Consequently, the total deposition percentage in the lower airways was independent of microsphere size and even 20 microm particles were able to penetrate into the lower airways, which was attributed to mouth breathing of the 1-day-old chickens. PMID- 17121738 TI - Infection and excretion of Salmonella Enteritidis in two different chicken lines with concurrent Ascaridia galli infection. AB - Studies on the impact of interaction of Salmonella enterica serovar Enteritidis and the parasitic nematode Ascaridia galli with the avian host were undertaken with particular emphasis on infection and excretion of these pathogens in two different layer lines. A total of 148 salmonella-free 1-day-old chickens (73 Hellevad and 75 Lohmann Brown) were randomly divided into five groups for each line. Group 1 served as an uninoculated control group. Groups 2 and 3 were infected with A. galli and S. Enteritidis, respectively. Group 4 was first infected with S. Enteritidis and subsequently with A. galli, and vice versa for group 5. The number of chickens excreting S. Enteritidis was significantly higher (P < 0.001) in the groups infected with both S. Enteritidis and A. galli compared with those only infected with S. Enteritidis over time. Furthermore, excretion of S. Enteritidis over time was significantly higher (P < 0.001) in the group first infected with S. Enteritidis and subsequently with A. galli compared with the group infected in the reverse order. No significant differences were observed between the two lines concerning excretion of S. Enteritidis over time in any group (P = 0.61 (group 3), P = 0.73 (group 4), P = 0.31 (group 5)). A. galli established itself significantly better (P = 0.02) in the group first infected with A. galli and subsequently with S. Enteritidis compared with the group infected in the reverse order. Furthermore, the A. galli infection rate was significantly higher (P = 0.02) in Hellevad chickens compared with Lohmann Brown chickens at the end of the experiment. PMID- 17121739 TI - Serological diagnosis of goose circovirus infections. AB - Infections with goose circovirus (GoCV) are associated with growth retardation and developmental problems in farmed geese. An indirect immunofluorescence assay for detecting virus-specific serum antibody was developed for diagnostic and epidemiological purposes. In the absence of a method for growing GoCV in cell culture, the assay was based on the reaction of antibodies with the GoCV capsid protein produced within baby hamster kidney cells using the eukaryotic Semliki forest virus expression vector. Using an optimized test that involved screening sera at 1:50 dilution and the use of a fluorescein isothiocyanate anti-duck immunoglobulin conjugate, GoCV-specific antibody was detected in 141 (88.6%) of 159 samples obtained from 27 of 28 breeder flocks aged from 1 to 6 years. Testing also showed the presence of GoCV-specific antibody in 85 (40.9%) of 208 serum samples from birds aged 30 weeks or less. Although maternally derived antibody was detected in birds when 1 and 4 days old, actively acquired antibody was first detected in birds aged 53 days. Following experimental inoculation of 21-day-old geese with tissue homogenate containing GoCV, virus-specific antibody was detected in serum samples collected at 27 and 34 days post inoculation. It is concluded that the SFV expression vector approach may prove useful for developing serological tests for other viruses, including other avian circoviruses, that do not grow in cell culture. PMID- 17121741 TI - Area social characteristics and carotid atherosclerosis. AB - OBJECTIVES: To explore the effect of social characteristics of residential areas on carotid atherosclerosis prevalence. METHODS AND RESULTS: The associations among area social characteristics and B-mode ultrasound determined carotid plaque score (a semi-quantitative scale measuring the degree of atherosclerosis in the carotid bifurcation area) were cross-sectionally investigated in a general population sample of 4033 men and women. Area socioeconomic circumstances were described through a social deprivation index calculated from migration rate, percentage residents with foreign citizenship among those with foreign background, dependency on social welfare support, and employment rate. Living in socially deprived areas was associated with an increased carotid plaque-score in both men (P for trend = 0.004) and women (P for trend = 0.007). These associations were only slightly reduced after adjustment for individual level indicators with a decrease of the absolute mean difference in carotid plaque score between worse-off and better-off areas of 9% for men and 13% for women, whereas adjustment for risk factors turned the trend non-significant in women, however, not in men. CONCLUSIONS: Those living in socially deprived areas in general had more extensive carotid atherosclerosis. However, in these areas there were a substantial number of individuals with low degrees of carotid atherosclerosis and vice versa. Thus, with regard to conceptual ideas of causal inference, the social characteristics of an area seem to be associated with the prevalence of carotid atherosclerosis. However, with regard to benefits of prevention, focusing on geographical areas would probably give a restricted benefit, where only some high-risk individuals would be reached. PMID- 17121742 TI - Contrasting acute and slow-growing lesions: a new door to brain plasticity. AB - The concept of plasticity describes the mechanisms that rearrange cerebral organization following a brain injury. During the last century, plasticity has been mainly investigated in humans with acute strokes. It was then shown: (i) that the brain is organized into highly specialized functional areas, often designated 'eloquent' areas and (ii) that a lesion within the eloquent area gives rise to major irrevocable deficits. However, in sharp contrast with these observations, it was recently found that patients with low-grade gliomas were able to undergo massive cerebral resections without detectable functional consequence. In this paper, we tackle this puzzling observation and address the idea that brain plasticity cannot be fully understood and fruitfully studied without considering the temporal pattern of the injury inflicted to the brain. To achieve this goal, we first review experimental evidence showing that functional recovery is considerably better in the context of slow-growing injuries than after acute lesions. Both human and animal data are considered. In a second step, we emphasize that slow and acute lesions involve very different patterns of reorganization. In agreement with this idea, we show that the recruitment of remote brain areas in the ipsi- and contralesional hemispheres is much more efficient in slow growing than acute lesions. Finally in a last section, we briefly discuss the main implications of these results. PMID- 17121743 TI - Spreading photoparoxysmal EEG response is associated with an abnormal cortical excitability pattern. AB - Photosensitivity or photoparoxysmal response (PPR) is a highly heritable electroencephalographic trait characterized by an abnormal cortical response to intermittent photic stimulation (IPS). In PPR-positive individuals, IPS induces spikes, spike-waves or intermittent slow waves. The PPR may be restricted to posterior visual areas (i.e. local PPR with occipital spikes only) or spread to anterior non-visual cortical regions (i.e. PPR with propagation). The mechanisms underlying the PPR and causing its spread remain to be clarified. In unmedicated PPR-positive individuals and PPR-negative control participants without any history of previous seizures, we used focal transcranial magnetic stimulation (TMS) to investigate the excitability of the visual or primary motor cortex (M1). In the first experiment [18 healthy control subjects (i.e. without PPR in electroencephalography: 6 females, mean age 26.5 +/- 7.34 years) and 17 healthy participants with PPR (7 females, mean age 25.18 +/- 12.2 years) were studied], occipital TMS was used to elicit phosphenes or to suppress the visual perception of letter trigrams. PPR-positive individuals with propagation had lower phosphene thresholds and steeper stimulus-response curves than individuals without PPR or with occipital spikes only. Occipital TMS also induced a stronger suppression of visual perception in PPR-positive subjects with propagation relative to subjects without PPR or with occipital spikes. In the second experiment, we applied TMS over the right M1 without concurrent IPS and measured the motor threshold, the stimulus response curve, and the duration of the cortical silent period (CSP) in PPR positive individuals with propagation and in PPR-negative control participants [15 right-handed healthy subjects without PPR (3 males, mean age 17.7 +/- 3.6 years) and 14 right-handed healthy individuals showing a PPR with propagation (3 males, mean age 17.4 +/- 3.9 years)]. PPR-positive individuals showed no changes in these excitability measures relative to the PPR-negative control participants. We also measured the modifiability of the CSP by continuous IPS at a frequency of 18 or 50 Hz. While IPS reduced the duration of the CSP in PPR-negative control subjects, IPS had no effect on the duration of the CSP in PPR-positive individuals. Our results provide first time evidence that the propagation of the PPR is associated with increased excitability of the occipital but not the motor cortex. The stronger inhibitory effect of TMS on visual perception and the failure of IPS to shorten the CSP in PPR-positive participants may possibly reflect adaptive changes that prevent the provocation of seizures during the PPR. PMID- 17121744 TI - TGF-beta receptor-mediated albumin uptake into astrocytes is involved in neocortical epileptogenesis. AB - It has long been recognized that insults to the cerebral cortex, such as trauma, ischaemia or infections, may result in the development of epilepsy, one of the most common neurological disorders. Human and animal studies have suggested that perturbations in neurovascular integrity and breakdown of the blood-brain barrier (BBB) lead to neuronal hypersynchronization and epileptiform activity, but the mechanisms underlying these processes are not known. In this study, we reveal a novel mechanism for epileptogenesis in the injured brain. We used focal neocortical, long-lasting BBB disruption or direct exposure to serum albumin in rats (51 and 13 animals, respectively, and 26 controls) as well as albumin exposure in brain slices in vitro. Most treated slices (72%, n = 189) displayed hypersynchronous propagating epileptiform field potentials when examined 5-49 days after treatment, but only 14% (n = 71) of control slices showed similar responses. We demonstrate that direct brain exposure to serum albumin is associated with albumin uptake into astrocytes, which is mediated by transforming growth factor beta receptors (TGF-betaRs). This uptake is followed by down regulation of inward-rectifying potassium (Kir 4.1) channels in astrocytes, resulting in reduced buffering of extracellular potassium. This, in turn, leads to activity-dependent increased accumulation of extracellular potassium, resulting in facilitated N-methyl-d-aspartate-receptor-mediated neuronal hyperexcitability and eventually epileptiform activity. Blocking TGF-betaR in vivo reduces the likelihood of epileptogenesis in albumin-exposed brains to 29.3% (n = 41 slices, P < 0.05). We propose that the above-described cascade of events following common brain insults leads to brain dysfunction and eventually epilepsy and suggest TGF-betaRs as a possible therapeutic target. PMID- 17121745 TI - What clinical disorders tell us about the neural control of saccadic eye movements. AB - Saccades are rapid eye movements that redirect the fovea from one object to another. A great deal has been learned about the anatomy and physiology of saccades, making them an ideal system for studying the neural control of movement. Basic research on normal eye movements has greatly increased our understanding of saccadic performance, anatomy and physiology, and led to a large number of control system models. These models simulate normal saccades well, but are challenged by clinical disorders because they often do not incorporate the specific anatomical and physiological substrates needed to model clinically important abnormalities. Historically, studies of saccadic abnormalities in patients have played a critical role in understanding the neural control of saccades because they provide information that complements basic research and thus restricts hypotheses to those that are biologically plausible. This review presents four examples of clinical disorders (slow saccades, interrupted saccades, high-frequency saccadic oscillations and macrosaccadic oscillations) that have provided insights into the neurobiology of saccades, have driven the development of new models, and have suggested an explanation or treatment for these disorders. We raise general questions for both scientists and clinicians that will assist in their efforts to understand the neural control of movement, improve diagnostic criteria and develop new treatments. PMID- 17121746 TI - Cortical activity in Parkinson's disease during executive processing depends on striatal involvement. AB - Patients with idiopathic Parkinson's disease exhibit impairments in executive processes, including planning and set-shifting, even at the early stages of the disease. We have recently developed a new card-sorting task to study the specific role of the caudate nucleus in such executive processes and have shown, using functional magnetic resonance imaging (fMRI) in young healthy adults, that the caudate nucleus is specifically required when a set-shift must be planned. Here the same fMRI protocol was used to compare the patterns of activation in a group of early-stage Parkinson's disease patients (seven right-handed patients at Hoehn and Yahr stages 1 and 2; mean age 62 years, range 56-70) and matched control subjects. Increased cortical activation was observed in the patients compared with the control group in the condition not specifically requiring the caudate nucleus. On the other hand, decreased cortical activation was observed in the patient group in the condition significantly involving the caudate nucleus. This event-related fMRI study showed a pattern of cortical activation in Parkinson's disease characterized by either reduced or increased activation depending on whether the caudate nucleus was involved or not in the task. This activation pattern included not only the prefrontal regions but also posterior cortical areas in the parietal and prestriate cortex. These findings are not in agreement with the traditional model, which proposes that the nigrostriatal dopamine depletion results in decreased cortical activity. These observations provide further evidence in favour of the hypothesis that not only the nigrostriatal and but also the mesocortical dopaminergic substrate may play a significant role in the cognitive deficits observed in Parkinson's disease. PMID- 17121747 TI - Use of Traditional Chinese Medicine by older Chinese immigrants in Canada. AB - BACKGROUND: Research is needed about the usage of complementary and alternative medicines within culturally diverse groups because of a growing number of people who use these remedies. OBJECTIVE: To understand the prevalence and predictors of Traditional Chinese Medicine (TCM) use by older Chinese immigrants in Canada. METHODS: This is based on the data collected from a representative sample of 2167 elderly Chinese immigrants aged 55 years and above in seven Canadian cities. Logistic regression was used to estimate the probability of using TCM in combination with Western health services (WHS). Use of Chinese herbs, herbal formulas, and TCM practitioners (herbalists) was predicted, based upon the effects of predisposing, enabling and need factors. RESULTS: The response rate was 77%. Over two-thirds of the older Chinese immigrants reported using TCM in combination with WHS. About half (50.3%) of the older Chinese immigrants used Chinese herbs, 48.7% used Chinese herbal formulas, and 23.8% consulted a Chinese herbalist. Although separate analysis was conducted, similar predictors were identified. Country of origin, Chinese health beliefs, social support, city of residency, and health variables were the common predictors of using a form of TCM. CONCLUSION: The combined use of TCM and WHS is common among elderly Chinese immigrants. Culture-related variables are important in determining use of TCM. The predictors identified should help physicians to recognize who among the elderly Chinese immigrants are more likely to use TCM so that a more in-depth understanding toward their health practices and needs can be achieved. PMID- 17121748 TI - Estrogen receptor alpha-351 XbaI*G and -397 PvuII*C-related genotypes and alleles are associated with higher susceptibilities of endometriosis and leiomyoma. AB - Endometriosis and leiomyoma are both common estrogen-related gynaecological diseases. We aimed to elucidate the association of estrogen receptor alpha (ERalpha)-351 A>G (XbaI) and -397 T>C (PvuII) gene polymorphisms with endometriosis and leiomyoma. Women were divided into three groups: (i) severe endometriosis (n = 112), (ii) leiomyoma (n = 106) and (iii) normal controls (n = 110). Genomic DNA was obtained from peripheral leukocytes. ERalpha-351 A/G XbaI and -397 T/C PvuII polymorphisms were assayed by the method of PCR and restriction fragment length polymorphism (RFLP). Genotypes and allelic frequencies in each group were compared. The genotype/allele frequencies of ERalpha-351 and -397 polymorphisms in endometriosis or leiomyoma groups were different from those of normal controls. ERalpha mutant-related genotypes/alleles (-351G and -397C) presented higher percentages in the endometriosis/leiomyoma population compared with normal controls. Proportions of ERalpha-351 AA/AG/GG genotypes and A/G alleles in each group were (i) 26.8/57.1/16.1 and 55.4/44.6%; (ii) 19.8/52.8/27.4 and 46.2/53.8% and (iii) 33.6/64.6/1.8 and 65.9/34.1%. Proportions of ERalpha-397 TT/TC/CC genotypes and T/C alleles in each group were (i) 24.1/60.7/15.2 and 54.5/45.5%; (ii) 23.6/70.8/5.6 and 59/41% and (iii) 54.5/40/5.5 and 74.5/25.5%. We concluded that ERalpha-351 XbaI*G- and -397 PvuII*C-related genotypes/alleles were correlated with higher susceptibilities of endometriosis or leiomyoma, which might be associated with related pathogeneses. PMID- 17121749 TI - Soluble HLA-G promotes Th1-type cytokine production by cytokine-activated uterine and peripheral natural killer cells. AB - Soluble forms of HLA-G (sHLA-G) have been implicated in immune regulation. Fetal trophoblast cells are a prime source of HLA-G. Hence, an interaction between sHLA G and uterine lymphocytes in the decidual tissues can easily be envisaged. These lymphocytes, when properly activated, are implicated in successful trophoblast invasion, placental maturation and maintenance of pregnancy. However, so far, no data are available on the effect of sHLA-G on the function and phenotype of these cells. Herein, we used a recombinant sHLA-G construct to determine the effect of sHLA-G on uterine lymphocyte cells present in endometrium at the time that it is optimally receptive to trophoblast invasion. In addition, we ascertained the effect of sHLA-G on peripheral lymphocytes. We found that upon co-culture with sHLA-G, proliferation of unfractionated IL-15-stimulated uterine mononuclear cells (UMCs) was inhibited. However, sHLA-G increased both interferon (IFN)-gamma and tumour necrosis factor (TNF)-alpha production by these cells. Vascular endothelial growth factor (VEGF) production was reduced. Notably, in contrast to membrane-bound HLA-G, sHLA-G did not affect the natural cytolytic activity of UMCs. Similarly, sHLA-G inhibited proliferation but stimulated pro-inflammatory cytokine production by cytokine-activated, unfractionated peripheral blood mononuclear cells (PBMCs). In addition, we showed that the overall inhibitory effect of sHLA-G on proliferation of the whole cell population could be ascribed to selective inhibition of CD4(+) T cells. In contrast, sHLA-G induced proliferation and IFN-gamma production by both uterine and peripheral natural killer (NK) cells. In conclusion, our data show that the sHLA-G modulates both UMC and PBMC function. sHLA-G, by promoting IFN-gamma production by uterine NK cells, may contribute to vascular remodelling of spiral arteries to allow for successful embryo implantation. PMID- 17121750 TI - Alcohol intake and incidence of coronary disease in Australian aborigines. AB - AIMS: To examine risk for coronary heart disease (CHD) and cardiovascular disease (CVD) in relation to alcohol in a cohort of Australian Aborigines. METHODS: In 1988-1989, alcohol intake, drinking pattern, and beverage preference were elicited by interviewer-administered questionnaire in Western Australian Aborigines (258 men and 256 women) and cardiovascular outcomes ascertained through linkage to mortality and hospital admission records to 2002. RESULTS: In proportional hazards models, risk for CHD, relative to lifetime abstainers, was significantly increased in ex-drinkers [Hazard ratio (HR), 2.29; 95% confidence intervals (CI), 1.23-4.27], those drinking 41-60 g/day in men or 21-40 g/day in women (HR 2.80; 95% CI, 1.04-7.53) and those drinking >150 g/day for men or >100 g/day for women (HR, 2.25; 95% CI, 1.03-4.90) with a J-shaped relationship. Low to-moderate drinkers had lower waist girth, exercised more and had a lower prevalence of overweight and smoking than at-risk drinkers. A preference for wine was associated with lower HR (0.28; 95% CI, 0.10-0.95). With CVD, only ex drinkers showed significantly increased risk (HR, 1.87; 95% CI, 1.20-2.91). CONCLUSIONS: More favourable health-related behaviours in low-to-moderate drinkers suggest that lower risk could be mediated by lifestyle, as proposed in other populations. PMID- 17121751 TI - Left ventricular outflow tract obstruction and sudden death in hypertrophic cardiomyopathy. PMID- 17121753 TI - QRS duration alone to select patients for cardiac resynchronization therapy: flying in the face of the evidence? PMID- 17121754 TI - Electrical storm: still a cryptogenic phenomenon? PMID- 17121755 TI - Value of cardiovascular magnetic resonance for determining cardiac involvement in systemic amyloidosis. PMID- 17121756 TI - Inhibition of CETP as a novel therapeutic strategy for reducing the risk of atherosclerotic disease. AB - Lowering low-density lipoprotein cholesterol (LDL-C) levels with statins is a proven strategy for reducing the risk of atherothrombotic cardiovascular disease (CVD). Yet, despite the success of statins in reducing cardiovascular event rates in at-risk patients, many will still experience further events. There is, therefore, a need to develop suitable therapies to reduce this residual risk. Low high-density lipoprotein cholesterol (HDL-C) levels are an important independent risk factor for CVD. Though fibrates, niacin, and statins have been shown to modestly raise HDL-C, there is increasing recognition of the need to develop therapies that can increase HDL-C more robustly. Such therapies may help supplement the LDL-C-lowering benefits of statins. Inhibition of cholesteryl ester transfer protein (CETP) has been identified as a possible strategy for substantially increasing HDL-C levels and CETP inhibitors have demonstrated clinical efficacy, in terms of increasing HDL-C, in preliminary clinical trials, and clinical trials based on outcomes are ongoing. Two CETP inhibitors, JTT-705 and torcetrapib, are now being evaluated more extensively. PMID- 17121757 TI - Impact of viability and scar tissue on response to cardiac resynchronization therapy in ischaemic heart failure patients. AB - AIMS: At present, 20-30% of patients do not respond to cardiac resynchronization therapy (CRT). In this study, the relation between the extent of viable myocardium and scar tissue vs. response to CRT was evaluated. In addition, the presence of scar tissue in the left ventricular (LV) lead position was specifically related to response to CRT. METHODS AND RESULTS: A total of 51 consecutive patients with ischaemic heart failure and substantial LV dyssynchrony undergoing CRT were included. All patients underwent gated SPECT before CRT implantation to determine the extent of scar tissue and viable myocardium. Clinical and echocardiographic parameters were assessed at baseline and after 6 months of CRT. The results demonstrated direct relations between the response to CRT and the extent of viable myocardium and scar tissue. In addition, the 15 patients (29%) with transmural scar tissue (< 50% tracer activity) in the region of the LV pacing lead showed no improvement after 6 months of CRT. CONCLUSION: The extent of scar tissue and viable myocardium were directly related to the response to CRT. Furthermore, scar tissue in the LV pacing lead region may prohibit response to CRT. Evaluation for viability and scar tissue may be considered in the selection process for CRT. PMID- 17121758 TI - Craniospinal irradiation using a forward planned segmented field technique. AB - Craniospinal irradiation is technically demanding due to the complex shape of the planning target volume (PTV). Radiotherapy treatment techniques have evolved over time as imaging and radiotherapy treatment technology have improved. However, most are variations on a class solution utilizing a prone patient position with two shaped lateral cranial portals and a matched posterior spinal portal with moving junctions. Major areas of difficulty remain with the accurate definition of the PTV and achieving a homogeneous dose within it, especially at the junctions. We describe a three-dimensionally (3D) planned craniospinal radiation technique that permits rapid image acquisition with reduced localization time, simplified spinal PTV definition and standardized cranial PTV definition. Improved dose homogeneity within the PTV is achieved by use of a segmented "field in-field" technique (forward planned intensity-modulated radiotherapy (IMRT)) in place of customized compensators. This has negated the requirement for constructing physical compensators. Autosequencing for field delivery enables the junction to be "moved" during a single fraction and reduces the overall treatment time, an important consideration when treating very young patients. PMID- 17121759 TI - A feasibility study of using gold seeds as fiducial markers for bladder localization during radical radiotherapy. AB - Target localization and verification of the treatment position is important for the accurate delivery of conformal radiotherapy. The bladder in particular is a deformable structure whose shape and position continually varies throughout a course of radiation treatment as a result of bladder filling. We report a novel technique of organ localization using gold seeds as fiducial markers that are implanted into the bladder using a specially adapted applicator that is passed through a rigid cystoscope. The seeds are readily apparent on electronic portal imaging taken at the time of radiotherapy and can thus act as a surrogate for bladder position. The feasibility and technical aspects of performing such a procedure on eight patients were assessed. In all of the patients, some of the seeds were visible on the planning CT scan and remained within the bladder wall throughout the course of radiotherapy treatment. The drop-out rate was minimized by the use of cystodiathermy at the site of seed insertion. It was possible to place the seeds in both areas of normal and diseased bladder tissue. The procedure was associated with minimal toxicity. This technique will form the basis for planning further studies on bladder localization. PMID- 17121760 TI - 123I-IMT SPECT for evaluation of the response to radiation therapy in high grade gliomas: a feasibility study. AB - Assessing response to radiation therapy in patients with high grade gliomas is needed upon making decisions toward further therapy strategies. Currently used standard imaging tools such as CT and MRI are not sensitive enough to detect early therapy effects. We prospectively investigated if single photon emission computed tomography (SPECT) using radiolabelled amino acid derivate (123)I methyltyrosine (IMT) would be useful for this aim. 10 patients with histologically proven high grade gliomas, who were candidates for radiation therapy, were enrolled in this investigation. All patients were examined by IMT SPECT before radiation therapy and 4 weeks after the initiation of the hypofractionated application of 40 Gy. Patients were followed up for 39 months; the tumours to background ratios (T/B) for IMT under/before radiation therapy were correlated to survival times. Initially, SPECT depicted an abnormal intratumoural IMT uptake in all patients (mean T/B ratios 1.37-1.87). In four out of 10 patients, the mean T/B ratios decreased by more than 10% under radiation therapy. In six other patients, the BQ decreased by less than 10% or increased. There were no significant correlations between the degree of changes in T/B and survival (r = -0.1, p = 0.973). Serial IMT SPECT measurements allow detection of changes in amino acid accumulation in high-grade gliomas under radiation therapy. However, these changes seem to possess no prognostic value in respect to survival prediction. PMID- 17121761 TI - A cost analysis of Behcet's syndrome in Turkey. AB - OBJECTIVE: Behcet's syndrome (BS), a common chronic inflammatory disease in Turkey, results in considerable morbidity and increased mortality. We aimed to estimate its direct and indirect costs. METHODS: We studied 119 (78 male/41 female) patients with BS at our multidisciplinary BS out-patient clinic in Istanbul, between March and July 2005. The mean age and disease duration were 35 +/- 9 and 10 +/- 6 S.D. yrs. According to the primary clinical problem, patients were divided into: (i) ocular; (ii) vascular; (iii) neurological and (iv) mucocutaneous-joint disease subgroups. They were interviewed with a standardized questionnaire addressing: (a) direct costs such as medication, diagnostic tests, hospital visits, hospitalization fees and lodging and transportation expenses and (b) indirect costs such as lost workdays and wages. The costs were assessed retrospectively by assessing the expenses within a year of the survey and were expressed as US$ mean +/- S.D. RESULTS: The mean annual total cost per patient was US$ 3226 +/- 3488 (S.D.). The direct cost accounted for 68% of the total cost. Medication expenses made up 79% of the total direct cost. About 42% of the patients reported lost workdays that averaged 119 +/- 96 days S.D. Among the clinical subgroups mucocutaneous-joint involvement (US$ 1180 +/- 1053) had the lowest economic impact while the neurological disease (US$ 5005 +/- 2707) had the highest. CONCLUSION: In this first cost of illness study in BS, the neurological involvement had the highest annual cost. Direct costs were higher than the indirect costs. Drug costs were the major cost driver. BS causes considerable economic burden for the health care system in Turkey. PMID- 17121762 TI - Diagnostic accuracy of a point-of-care syphilis test when used among pregnant women in Bolivia. AB - OBJECTIVE: To evaluate the performance of a point-of-care (POC) syphilis test when used in urban Bolivian maternity hospitals. METHODS: We tested 8892 pregnant women for syphilis using the Abbott Determine Syphilis TP rapid POC test and rapid plasma reagin (RPR) in the laboratory of four large urban maternity hospitals where national statistics reported a syphilis prevalence of at least 3%. Sera were stored and transferred to the national reference laboratory (INLASA) where RPR testing was repeated. When the reference laboratory staff observed a positive RPR result, a Treponema pallidum particle agglutination assay (TPPA) was performed to confirm these findings. We calculated test performance characteristics for the POC test and hospital RPR using RPR performed at the reference laboratory confirmed by TPPA as the reference standard. Participants received treatment during their initial visit based on the POC test results. RESULTS: The sensitivity, specificity, negative predictive value and positive predictive values of the POC syphilis test were: 91.8% (95% confidence intervals 88.4% to 94.5%), 98.5% (98.2% to 98.8%), 71.0% (66.6% to 75.2%), and 99.7% (99.5% to 99.8%), respectively. The RPR values were 75.7% (70.8% to 80.2%), 99.0% (98.9% to 99.3%), 76.9% (72.0% to 81.3%), and 99.0% (98.8% to 99.2%), respectively. CONCLUSION: The Abbott Determine Syphilis TP test proved to be more sensitive than routine RPR and had comparable specificity. POC testing may be a simple way to expand syphilis screening to clinics with no laboratory facilities, improve case detection, and facilitate treatment delivery. PMID- 17121763 TI - Clinic-based evaluation of Clearview Chlamydia MF for detection of Chlamydia trachomatis in vaginal and cervical specimens from women at high risk in China. AB - OBJECTIVES: To determine the performance of a rapid Chlamydia trachomatis (CT) test (Clearview Chlamydia MF) compared to the current "gold standard" (Roche Amplicor CT assay) test, and to assess acceptability of the tests to patients. METHODS: A total of 1497 women at sexually transmitted diseases (STD) clinics or re-education centres in six urban cities (Shanghai, Nanjing, Shenzhen, Guangzhou, Chengdu and Fuzhou) in China participated in the study. Three vaginal and three cervical swabs were collected from each participant. Rapid CT tests were performed locally on the first vaginal and cervical swabs and the results were read independently by two staff members. The second and third swabs were randomised for performing the Roche CT assay at the National STD Reference Laboratory. Acceptability of the rapid tests to patients was determined by asking patients in clinics about their willingness to wait for the results. RESULTS: The prevalence of CT was 13.2% (197/1497), as determined by the Roche assay with cervical specimens. CT was detected in 78 vaginal and 127 cervical specimens by the rapid test and the positive rates determined with cervical specimens were significantly higher than those with vaginal specimens (p<0.001). There was good agreement between the results read by two independent staff for either vaginal or cervical specimens (both kappa = 0.98, p<0.001). Sensitivities for vaginal and cervical specimens were 32.8% and 49.7%, respectively, and specificities were 99.2% and 97.9%, respectively. The positive predictive value was 85.7% for vaginal and 78.4% for cervical specimens. The vast majority of the patients (99.1%) were willing to wait up to two hours for the results. CONCLUSION: Clearview Chlamydia MF, while yielding a rapid result and requiring minimal laboratory facilities, had unacceptably low sensitivity compared to a nucleic acid amplification test. Rapid tests yielding results within one hour are generally accepted by the clients. PMID- 17121764 TI - Diagnosing tuberculous pericarditis. AB - BACKGROUND: Definitive diagnosis of tuberculous pericarditis requires isolation of the tubercle bacillus from pericardial fluid, but isolating the organism is often difficult. AIM: To improve diagnostic efficiency for tuberculous pericarditis, using available tests. DESIGN: Prospective observational study. METHODS: Consecutive patients (n = 233) presenting with pericardial effusions underwent a predetermined diagnostic work-up. This included (i) clinical examination; (ii) pericardial fluid tests: biochemistry, microbiology, cytology, differential white blood cell (WBC) count, gamma interferon (IFN-gamma), adenosine deaminase (ADA) levels, polymerase chain reaction testing for Mycobacterium tuberculosis; (iii) HIV; (iv) sputum smear and culture; (v) blood biochemistry; and (vi) differential WBC count. A model was developed using 'classification and regression tree' analysis. The cut-off for the total diagnostic index (DI) was optimized using receiver operating characteristic (ROC) curves. RESULTS: Fever, night sweats, weight loss, serum globulin (>40 g/l) and peripheral blood leukocyte count (<10 x 10(9)/l) were independently predictive. The derived prediction model had 86% sensitivity and 84% specificity when applied to the study population. Pericardial fluid IFN-gamma >or=50 pg/ml, concentration had 92% sensitivity, 100% specificity and a positive predictive value (PPV) of 100% for the diagnosis of tuberculous pericarditis; pericardial fluid ADA >or=40 U/l had 87% sensitivity and 89% specificity. A diagnostic model including pericardial ADA, lymphocyte/neutrophil ratio, peripheral leukocyte count and HIV status had 96% sensitivity and 97% specificity; substituting pericardial IFN gamma for ADA yielded 98% sensitivity and 100% specificity. DISCUSSION: Basic clinical and laboratory features can aid the diagnosis of tuberculous pericarditis. If available, pericardial IFN-gamma is the most useful diagnostic test. Otherwise we propose a prediction model that incorporates pericardial ADA and differential WBC counts. PMID- 17121765 TI - Bromelain as an adjunctive treatment for moderate-to-severe osteoarthritis of the knee: a randomized placebo-controlled pilot study. AB - BACKGROUND: Osteoarthritis (OA) of the knee is the most prevalent joint disorder. Previous studies suggest that bromelain, a pineapple extract, may be a safer alternative/adjunctive treatment for knee OA than current conventional treatment. AIM: To assess the efficacy of bromelain in treating OA of the knee. DESIGN: Randomized, double-blind placebo-controlled trial. METHODS: Subjects (n = 47) with a confirmed diagnosis of moderate to severe knee OA were randomized to 12 weeks of bromelain 800 mg/day or placebo, with a 4-week follow-up. Knee (pain, stiffness and function) and quality-of-life symptoms were reported monthly in the WOMAC and SF36 questionnaires, respectively. Adverse events were also recorded. The primary outcome measure was the change in total WOMAC score from baseline to the end of treatment at week 12. Longitudinal models were used to evaluate outcome. RESULTS: Thirty-one patients completed the trial (14 bromelain, 17 placebo). No statistically significant differences were observed between groups for the primary outcome (coefficient 11.16, p = 0.27, 95%CI -8.86 to 31.18), nor the WOMAC subscales or SF36. Both treatment groups showed clinically relevant improvement in the WOMAC disability subscale only. Adverse events were generally mild in nature. DISCUSSION: This study suggests that bromelain is not efficacious as an adjunctive treatment of moderate to severe OA, but its limitations support the need for a follow-up study. PMID- 17121766 TI - Abnormal haemoglobin levels in acute coronary syndromes. AB - BACKGROUND: Anaemia is an adverse prognostic marker in acute coronary syndromes (ACS), but the epidemiology of abnormal haemoglobin levels in such patients is uncertain. AIMS: To investigate the prevalence, nature and predictors of abnormal haemoglobin levels in ACS patients at admission. DESIGN: Observational study. METHODS: All emergency admissions from January to April 2005 were assessed within 24-48 h of hospital admission. ACS patients (unstable angina, non-ST-elevation or ST-elevation myocardial infarction) were enrolled (n = 320, 190 men). Clinical information was recorded. RESULTS: Overall, 71% had unstable angina; 18% non-ST elevation myocardial infarction (MI), and 11% ST-elevation MI. Mean +/- SD haemoglobin was 14.3 +/- 1.7 g/dl in men and 13.2 +/- 1.5 g/dl in women. Abnormal haemoglobin was more common in men (65, 34%) than in women (34, 22%) (p = 0.013). Anaemia (haemoglobin <13 g/dl in men, or <12.0 g/dl in women) was recorded in 35 (18%) men and 24 (18%) women. All had admission haemoglobin >8 g/dl, and anaemia was usually normocytic. Multivariate predictors of anaemia (OR, 95%CI) were age (1.07, 1.04-1.1) and serum albumin (0.90, 0.81-1.00). Elevated haemoglobin (>16 g/dl) was recorded in 30 (16%) men and 4 (3%) women (p < 0.01), and was more common in ST-elevation MI patients (26%) than in unstable angina or non-ST elevation MI patients (9%) (p = 0.005). In patients who underwent invasive management with a post-procedure haemoglobin the following day (n = 85), 15 (18%) new cases of anaemia were detected. Admission duration correlated with haemoglobin (p < 0.01), creatinine (p < 0.01), troponin I (p < 0.01) and C reactive protein (p < 0.01). Anaemia was more common in those who died in hospital (3, 60%) than in those who survived (56, 18%) (adjusted p = 0.0135). DISCUSSION: Abnormal haemoglobin levels were common in our ACS admissions. Anaemia was generally mild. Increasing age and interventional management were associated with anaemia, which in turn was associated with adverse in-hospital outcomes. Interventions to prevent and detect anaemia in this setting merit prospective testing. PMID- 17121767 TI - Cerebral haemodynamics in acute bacterial meningitis in adults. AB - BACKGROUND: Vascular complications are an important cause of neurological sequelae among adult survivors of acute bacterial meningitis (ABM). AIM: To examine the haemodynamic changes associated with ABM. METHODS: Serial transcranial colour-coded sonography (TCCS) and magnetic resonance angiography (MRA) were used to examine cerebrovascular changes in adult ABM patients. Outcome at 3 months was categorized using a modified Barthel index. RESULTS: We recruited 24 patients, 12 men and 12 women, aged 21-68 years. Mean cerebral blood flow velocity (V(mean)) increased from day 1 to day 4 in the middle cerebral artery (MCA), anterior cerebral artery (ACA) and posterior cerebral artery (PCA). On day 4, V(mean) values in the MCA, ACA and PCA were all significantly higher than reference values in healthy volunteers. At 3 months follow-up, 16 cases had good outcomes, while the other eight had poor outcomes. Under multiple logistic regression analysis, only Glasgow coma score (GCS) at admission was independently associated with the three-month outcome. DISCUSSION: In these patient, stenosis as demonstrated by TCCS did not wholly coincide with stenosis as demonstrated by MRA, and the presence of intracranial stenosis was not predictive of a poor outcome at 3 months. Further studies are needed to delineate the characteristics and significance of cerebrovascular changes in adult ABM. PMID- 17121768 TI - Elevated serum alanine transaminase in patients with type 1 or type 2 diabetes mellitus. AB - BACKGROUND: The risk of chronic liver disease is higher in diabetics, and serum alanine transaminase (ALT) is a sensitive predictor of mortality from liver disease. AIM: To estimate the prevalence of elevated ALT in type 1 and type 2 diabetes, and identify possible risk factors. METHODS: We identified all patients (n = 2077) attending review between September 2002 and August 2003. We excluded those with no ALT measurement (n = 73); those whose alcohol consumption was >14 units/week (women) (n = 276) or >21 units/week (men) (n = 324); and those with a diagnosis of Maturity Onset Diabetes of the Young, secondary diabetes, gestational diabetes or uncertain type of diabetes (n = 51). We calculated the prevalences of elevated ALT in both type 1 and type 2 diabetes patients, and compared the demographic, microvascular risk factors and current drug use between each group using multivariate logistic regression. RESULTS: Of 1353 patients included, 836 (61.9%) had type 2 diabetes. Elevated ALT was found in 9.5% (95%CI 7.1-12.3%) of patients with type 1 diabetes, and 12.1% (95%CI 9.9-14.5%) of those with type 2 diabetes. The risk of elevated ALT in patients with type 2 diabetes increased with increasing body mass index (p(trend) = 0.04), and was lower in those taking insulin (OR 0.38, 95%CI 0.22-0.65). DISCUSSION: The prevalence of elevated ALT is 3-4 times higher in patients with either type 1 or type 2 diabetes than in the general population. Further studies investigating the aetiology and mechanisms of this elevation may suggest appropriate early interventions. PMID- 17121769 TI - Hyponatraemia as a risk factor for hospital mortality. AB - Hyponatraemia is a common abnormality in hospitalized patients, with about 15% having levels below the lower limit of the laboratory reference range. Accepted wisdom is that hyponatraemia is a marker of poor prognosis. However, a critical analysis of the literature reveals significant problems. Researchers have used various cut-off levels for plasma sodium, often concentrating on more severely hyponatraemic groups. Many studies were small, and most did not include control groups. Nevertheless, the literature available does suggest an excess mortality associated with hyponatraemia. Whether this is a direct adverse effect of low serum sodium levels, or if hyponatraemia is simply a marker for 'sicker' patients, is not known. It is also uncertain whether mortality is increased with more severe hyponatraemia, or whether active correction of hyponatraemia will improve outcome. These issues should be addressed by adequately-powered, prospective, suitably controlled studies. PMID- 17121771 TI - Treating metabolic acidosis. PMID- 17121773 TI - PrepMS: TOF MS data graphical preprocessing tool. AB - We introduce a simple-to-use graphical tool that enables researchers to easily prepare time-of-flight mass spectrometry data for analysis. For ease of use, the graphical executable provides default parameter settings, experimentally determined to work well in most situations. These values, if desired, can be changed by the user. PrepMS is a stand-alone application made freely available (open source), and is under the General Public License (GPL). Its graphical user interface, default parameter settings, and display plots allow PrepMS to be used effectively for data preprocessing, peak detection and visual data quality assessment. AVAILABILITY: Stand-alone executable files and Matlab toolbox are available for download at: http://sourceforge.net/projects/prepms PMID- 17121774 TI - Qualitatively modelling and analysing genetic regulatory networks: a Petri net approach. AB - MOTIVATION: New developments in post-genomic technology now provide researchers with the data necessary to study regulatory processes in a holistic fashion at multiple levels of biological organization. One of the major challenges for the biologist is to integrate and interpret these vast data resources to gain a greater understanding of the structure and function of the molecular processes that mediate adaptive and cell cycle driven changes in gene expression. In order to achieve this biologists require new tools and techniques to allow pathway related data to be modelled and analysed as network structures, providing valuable insights which can then be validated and investigated in the laboratory. RESULTS: We propose a new technique for constructing and analysing qualitative models of genetic regulatory networks based on the Petri net formalism. We take as our starting point the Boolean network approach of treating genes as binary switches and develop a new Petri net model which uses logic minimization to automate the construction of compact qualitative models. Our approach addresses the shortcomings of Boolean networks by providing access to the wide range of existing Petri net analysis techniques and by using non-determinism to cope with incomplete and inconsistent data. The ideas we present are illustrated by a case study in which the genetic regulatory network controlling sporulation in the bacterium Bacillus subtilis is modelled and analysed. AVAILABILITY: The Petri net model construction tool and the data files for the B. subtilis sporulation case study are available at http://bioinf.ncl.ac.uk/gnapn. PMID- 17121775 TI - SNP2NMD: a database of human single nucleotide polymorphisms causing nonsense mediated mRNA decay. AB - Elucidating the effects of genetic polymorphisms on genes and gene networks is an important step in disease association studies. We developed the SNP2NMD database for human SNPs (single nucleotide polymorphisms) that result in PTCs (premature termination codons) and trigger nonsense-mediated mRNA decay (NMD). The SNP2NMD Web interfaces provide extensive genetic information on and graphical views of the queried SNP, gene, and disease terms. AVAILABILITY: SNP2NMD is available from http://variome.net, or directly from http://bioportal.kobic.re.kr/SNP2NMD. SUPPLEMENTARY INFORMATION: http://bioportal.kobic.re.kr/SNP2NMD/Wiki.jsp?page=Statistics. PMID- 17121776 TI - ProteomeCommons.org IO Framework: reading and writing multiple proteomics data formats. AB - MOTIVATION: Effective use of proteomics data, specifically mass spectrometry data, relies on the ability to read and write the many mass spectrometer file formats. Even with mass spectrometer vendor-specific libraries and vendor-neutral file formats, such as mzXML and mzData it can be difficult to extract raw data files in a form suitable for batch processing and basic research. Introduced here are the ProteomeCommons.org Input and Output Framework, abbreviated to IO Framework, which is designed to abstractly represent mass spectrometry data. This project is a public, open-source, free-to-use framework that supports most of the mass spectrometry data formats, including current formats, legacy formats and proprietary formats that require a vendor-specific library in order to operate. The IO Framework includes an on-line tool for non-programmers and a set of libraries that developers may use to convert between various proteomics file formats. AVAILABILITY: The current source-code and documentation for the ProteomeCommons.org IO Framework is freely available at http://www.proteomecommons.org/current/531/ PMID- 17121777 TI - ARIA2: automated NOE assignment and data integration in NMR structure calculation. AB - Modern structural genomics projects demand for integrated methods for the interpretation and storage of nuclear magnetic resonance (NMR) data. Here we present version 2.1 of our program ARIA (Ambiguous Restraints for Iterative Assignment) for automated assignment of nuclear Overhauser enhancement (NOE) data and NMR structure calculation. We report on recent developments, most notably a graphical user interface, and the incorporation of the object-oriented data model of the Collaborative Computing Project for NMR (CCPN). The CCPN data model defines a storage model for NMR data, which greatly facilitates the transfer of data between different NMR software packages. AVAILABILITY: A distribution with the source code of ARIA 2.1 is freely available at http://www.pasteur.fr/recherche/unites/Binfs/aria2. PMID- 17121778 TI - QOMA: quasi-optimal multiple alignment of protein sequences. AB - MOTIVATION: We consider the problem of multiple alignment of protein sequences with the goal of achieving a large SP (Sum-of-Pairs) score. RESULTS: We introduce a new graph-based method. We name our method QOMA (Quasi-Optimal Multiple Alignment). QOMA starts with an initial alignment. It represents this alignment using a K-partite graph. It then improves the SP score of the initial alignment through local optimizations within a window that moves greedily on the alignment. QOMA uses two parameters to permit flexibility in time/accuracy trade off: (1) The size of the window for local optimization. (2) The sparsity of the K-partite graph. Unlike traditional progressive methods, QOMA is independent of the order of sequences. The experimental results on BAliBASE benchmarks show that QOMA produces higher SP score than the existing tools including ClustalW, Probcons, Muscle, T-Coffee and DCA. The difference is more significant for distant proteins. AVAILABILITY: The software is available from the authors upon request. PMID- 17121779 TI - A predictive model for identifying proteins by a single peptide match. AB - MOTIVATION: Tandem mass-spectrometry of trypsin digests, followed by database searching, is one of the most popular approaches in high-throughput proteomics studies. Peptides are considered identified if they pass certain scoring thresholds. To avoid false positive protein identification, > or = 2 unique peptides identified within a single protein are generally recommended. Still, in a typical high-throughput experiment, hundreds of proteins are identified only by a single peptide. We introduce here a method for distinguishing between true and false identifications among single-hit proteins. The approach is based on randomized database searching and usage of logistic regression models with cross validation. This approach is implemented to analyze three bacterial samples enabling recovery 68-98% of the correct single-hit proteins with an error rate of < 2%. This results in a 22-65% increase in number of identified proteins. Identifying true single-hit proteins will lead to discovering many crucial regulators, biomarkers and other low abundance proteins. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 17121780 TI - Rapidly progressive renal failure associated with successful pharmacotherapy for obesity. PMID- 17121781 TI - Rapid monitoring of bacteria in dialysis fluids by fluorescent vital staining and microcolony methods. PMID- 17121782 TI - Hyperpnoea and ketonuria in an HIV-infected patient. PMID- 17121783 TI - Kidney mass and osteolytic lesion: is it always malignancy? PMID- 17121784 TI - The association of higher depressive symptoms and sexual dysfunction in male haemodialysis patients. AB - BACKGROUND: The prevalence of sexual dysfunction among male haemodialysis patients is high. Sexual dysfunction is composed of both physiological and psychological factors. However, the role of psychological depression is still obscure. METHODS: A multicentre cross-sectional study of 411 male haemodialysis patients was conducted to define the determinants of sexual dysfunction. Mid-week pre-dialytic biochemical and haematological parameters were obtained. All patients were required to complete three questionnaires by themselves: (i) the International Index of Erectile Function (IIEF, Chinese version); (ii) the Beck Depression Inventory (BDI, Chinese version) and (iii) the 36-item Short Form Health Survey Questionnaire (SF-36, Taiwan Standard Version 1.0). RESULTS: In total, 154 male patients completed the IIEF questionnaire. Their mean age was 50.2 +/- 10.7 years. A linear multivariable regression analysis demonstrated advanced age, diabetes and the presence of depressive symptoms to be independently associated with sexual dysfunction. Subjects with sexual dysfunction had significantly lower quality of life scores. CONCLUSIONS: The presence of depressive symptoms, highly prevalent in haemodialysis patients, is an independent factor of sexual dysfunction in male haemodialysis patients. In a comprehensive approach to the management of sexual dysfunction, a thorough evaluation of psychological depression must be included. PMID- 17121785 TI - The haemodynamic and metabolic effects of hypertonic-glucose and amino-acid-based peritoneal dialysis fluids. AB - BACKGROUND: Continuous ambulatory peritoneal dialysis (CAPD) may exert significant effects on systemic haemodynamics. We have previously demonstrated that hypertonic glucose solutions are associated with higher blood pressure (BP) due to a rise in stroke volume (SV) and cardiac output (CO). However, the mechanisms underlying these changes have not been established. METHODS: Ten non diabetic CAPD patients entered a randomized crossover study (eight completed) to compare conventional glucose-based fluid, biocompatible pH-neutral glucose-based fluid and 1.1% amino acid solution (lactate-buffered but completely free of glucose degradation products). BP and haemodynamic variables were measured using continuous arterial pulse wave analysis, and serial plasma glucose and insulin concentrations were collected. Left ventricular (LV) diameters were measured at the start and end of each dwell period using M-mode echocardiography. RESULTS: BP was similar during 1.36% glucose and 1.1% amino acid dwells, but was significantly higher during 3.86% glucose dwells with both conventional and biocompatible fluids (P < 0.001). This was associated with a significantly higher SV and CO (P < 0.001), although the haemodynamic response differed between conventional and biocompatible 3.86% solutions. Plasma glucose and insulin levels did not differ from baseline during 1.36% and amino acid dwells, but increased significantly during 3.86% glucose dwells. Despite a significantly higher ultrafiltration volume with 3.86% glucose, LV diameters were similar throughout. CONCLUSIONS: In conclusion, we have confirmed our previous findings demonstrating higher BP and adverse haemodynamics during 3.86% glucose dwells. These changes are associated with hyperglycaemia and hyperinsulinaemia, but are not related to differences in cardiac filling. PMID- 17121786 TI - Association of the VEGF gene polymorphism with diabetic retinopathy in type 2 diabetes patients. AB - BACKGROUND: Diabetic microvascular complications are the major causes of morbidity and early mortality in diabetes. Vascular endothelial growth factor (VEGF) is a potent multifunctional cytokine which plays a key role in the pathogenesis of diabetic microvascular complications. We examined the possible association of the VEGF gene polymorphisms with diabetic nephropathy and retinopathy in type 2 diabetes patients. METHODS: Genotyping of the VEGF gene insertion/deletion (I/D) and +405 polymorphisms was done by the polymerase chain reaction (PCR) and restriction fragment length polymorphism methods. A total of 426 patients with type 2 diabetes and 493 healthy subjects were genotyped. The frequency of VEGF alleles and genotype distribution were compared in diabetic and control groups. RESULTS: The distribution of the VEGF DD genotype was significantly different in patients with diabetic retinopathy compared with healthy controls, entire diabetic group and patients with no complications (44 vs. 23, 30 and 21%, respectively; P < 0.01). Such differences were not observed in the diabetic nephropathy group. The odds ratio for the D allele was 2.27 (95% CI 1.59-3.25). The multivariate logistic regression analysis revealed that the D allele of the VEGF gene I/D polymorphism was an independent risk factor of retinopathy (P < 0.001). The VEGF +405 genotype was not associated with diabetic complications in type 2 diabetes patients. CONCLUSION: Our study suggests that the I/D polymorphism in the promoter region of the VEGF gene is associated with retinopathy but not nephropathy in type 2 diabetes patients. The multivariate logistic regression analysis showed that the D allele of the VEGF polymorphism is an independent risk factor of diabetic retinopathy after controlling for other clinical variables. PMID- 17121787 TI - Potential role of active vitamin D in retarding the progression of chronic kidney disease. PMID- 17121788 TI - Physiological levels of virion-associated human immunodeficiency virus type 1 envelope induce coreceptor-dependent calcium flux. AB - Human immunodeficiency virus (HIV) entry into target cells requires the engagement of receptor and coreceptor by envelope glycoprotein (Env). Coreceptors CCR5 and CXCR4 are chemokine receptors that generate signals manifested as calcium fluxes in response to binding of the appropriate ligand. It has previously been shown that engagement of the coreceptors by HIV Env can also generate Ca(2+) fluxing. Since the sensitivity and therefore the physiological consequence of signaling activation in target cells is not well understood, we addressed it by using a microscopy-based approach to measure Ca(2+) levels in individual CD4(+) T cells in response to low Env concentrations. Monomeric Env subunit gp120 and virion-bound Env were able to activate a signaling cascade that is qualitatively different from the one induced by chemokines. Env-mediated Ca(2+) fluxing was coreceptor mediated, coreceptor specific, and CD4 dependent. Comparison of the observed virion-mediated Ca(2+) fluxing with the exact number of viral particles revealed that the viral threshold necessary for coreceptor activation of signaling in CD4(+) T cells was quite low, as few as two virions. These results indicate that the physiological levels of virion binding can activate signaling in CD4(+) T cells in vivo and therefore might contribute to HIV-induced pathogenesis. PMID- 17121789 TI - Functional p53 signaling in Kaposi's sarcoma-associated herpesvirus lymphomas: implications for therapy. AB - The Kaposi's sarcoma-associated herpesvirus (KSHV/HHV8) is associated with Kaposi's sarcoma (KS) as well as primary effusion lymphomas (PEL). The expression of viral proteins capable of inactivating the p53 tumor suppressor protein has been implicated in KSHV oncogenesis. However, DNA-damaging drugs such as doxorubicin are clinically efficacious against PEL and KS, suggesting that p53 signaling remains intact despite the presence of KSHV. To investigate the functionality of p53 in PEL, we examined the response of a large number of PEL cell lines to doxorubicin. Two out of seven (29%) PEL cell lines harbored a mutant p53 allele (BCBL-1 and BCP-1) which led to doxorubicin resistance. In contrast, all other PEL containing wild-type p53 showed DNA damage-induced cell cycle arrest, p53 phosphorylation, and p53 target gene activation. These data imply that p53-mediated DNA damage signaling was intact. Supporting this finding, chemical inhibition of p53 signaling in PEL led to doxorubicin resistance, and chemical activation of p53 by the Hdm2 antagonist Nutlin-3 led to unimpaired induction of p53 target genes as well as growth inhibition and apoptosis. PMID- 17121790 TI - Adenovirus infection triggers a rapid, MyD88-regulated transcriptome response critical to acute-phase and adaptive immune responses in vivo. AB - Nearly 50 years ago, the discovery of interferon prompted the notion that host cells innately respond to viral invasion. Since that time, technological advances have allowed this response to be extensively characterized and dissected in vitro. However, these advances have only recently been applied to highly complex, in vivo biological systems. To this end, we exploited high-titer adenovirus (Ad) vectors to globally investigate the innate immune response to nonenveloped viral infection in vivo. Our results indicated a potent cellular transcriptome response shortly after infection, with global assessments revealing significant dysregulation in approximately 15% of the measured transcripts derived from Ad vector-transduced tissue. Bioinformatics-based transcriptome analysis revealed a complex innate response to Ad infection, with induction of proinflammatory responses (and suppression of metabolism and mitochondrial genes) akin to those observed when mice are challenged with lipopolysaccharide. Despite this commonality, there were many unique aspects of the Ad-dependent transcriptome response, including the upregulation of several RNA regulatory mechanisms and apoptosis-related pathways, accompanied by the suppression of lysosomal and endocytic genes. Our results also implicated the Toll-like receptors (TLRs) in these responses, prompting specific investigations into this pathway. By using MyD88KO mice, our results confirmed that Ad-induced dysregulation of five functionally related gene clusters are significantly dependent on this TLR adaptor gene. MyD88 deficiency also resulted in significantly diminished, although not abolished, adaptive and acute-phase immune responses to Ad, confirming the transcriptome data, as well as specifically identifying MyD88 as a significant Ad immunity amplifier and regulator in vivo. PMID- 17121791 TI - Inhibition of the secretory pathway by foot-and-mouth disease virus 2BC protein is reproduced by coexpression of 2B with 2C, and the site of inhibition is determined by the subcellular location of 2C. AB - Infection of cells with picornaviruses can lead to a block in protein secretion. For poliovirus this is achieved by the 3A protein, and the consequent reduction in secretion of proinflammatory cytokines and surface expression of major histocompatibility complex class I proteins may inhibit host immune responses in vivo. Foot-and-mouth disease virus (FMDV), another picornavirus, can cause persistent infection of ruminants, suggesting it too may inhibit immune responses. Endoplasmic reticulum (ER)-to-Golgi apparatus transport of proteins is blocked by the FMDV 2BC protein. The observation that 2BC is processed to 2B and 2C during infection and that individual 2B and 2C proteins are unable to block secretion stimulated us to study the effects of 2BC processing on the secretory pathway. Even though 2BC was processed rapidly to 2B and 2C, protein transport to the plasma membrane was still blocked in FMDV-infected cells. The block could be reconstituted by coexpression of 2B and 2C, showing that processing of 2BC did not compromise the ability of FMDV to slow secretion. Under these conditions, 2C was located to the Golgi apparatus, and the block in transport also occurred in the Golgi apparatus. Interestingly, the block in transport could be redirected to the ER when 2B was coexpressed with a 2C protein fused to an ER retention element. Thus, for FMDV a block in secretion is dependent on both 2B and 2C, with the latter determining the site of the block. PMID- 17121792 TI - The cytomegalovirus homolog of interleukin-10 requires phosphatidylinositol 3 kinase activity for inhibition of cytokine synthesis in monocytes. AB - Human cytomegalovirus (CMV) has evolved numerous strategies for evading host immune defenses, including piracy of cellular cytokines. A viral homolog of interleukin-10, designated cmvIL-10, binds to the cellular IL-10 receptor and effects potent immune suppression. The signaling pathways employed by cmvIL-10 were investigated, and the classic IL-10R/JAK1/Stat3 pathway was found to be activated in monocytes. However, inhibition of JAK1 had little effect on cmvIL-10 mediated suppression of tumor necrosis factor alpha (TNF-alpha) production. Inhibition of the phosphatidylinositol 3-kinase/Akt pathway had a more significant impact on TNF-alpha levels but did not completely relieve the immune suppression, demonstrating that cmvIL-10 stimulates multiple signaling pathways to modulate cell function. PMID- 17121793 TI - Mutually exclusive T-cell receptor induction and differential susceptibility to human immunodeficiency virus type 1 mutational escape associated with a two-amino acid difference between HLA class I subtypes. AB - The relative contributions of HLA alleles and T-cell receptors (TCRs) to the prevention of mutational viral escape are unclear. Here, we examined human immunodeficiency virus type 1 (HIV-1)-specific CD8(+) T-cell responses restricted by two closely related HLA class I alleles, B*5701 and B*5703, that differ by two amino acids but are both associated with a dominant response to the same HIV-1 Gag epitope KF11 (KAFSPEVIPMF). When this epitope is presented by HLA-B*5701, it induces a TCR repertoire that is highly conserved among individuals, cross recognizes viral epitope variants, and is rarely associated with mutational escape. In contrast, KF11 presented by HLA-B*5703 induces an entirely different, more heterogeneous TCR beta-chain repertoire that fails to recognize specific KF11 escape variants which frequently arise in clade C-infected HLA-B*5703(+) individuals. These data show the influence of HLA allele subtypes on TCR selection and indicate that extensive TCR diversity is not a prerequisite to prevention of allowable viral mutations. PMID- 17121794 TI - Microglial cell line established from prion protein-overexpressing mice is susceptible to various murine prion strains. AB - Several lines of evidence suggest that microglia have important roles in the pathogenesis of prion diseases. Here, we establish a novel microglial cell line (MG20) from neonatal tga20 mice that overexpress murine prion protein. After exposure to Chandler scrapie, we observed the replication and accumulation of disease-associated forms of the prion protein in MG20 cells up to the 15th passage. Furthermore, MG20 cells were susceptible to ME7, Obihiro scrapie, and bovine spongiform encephalopathy agents. Thus, MG20 cell lines persistently infected with various murine prion strains provide a useful model for the study of the pathogenesis of prion diseases. PMID- 17121795 TI - Proteasome-independent major histocompatibility complex class I cross presentation mediated by papaya mosaic virus-like particles leads to expansion of specific human T cells. AB - The development of versatile vaccine platforms is a priority that is recognized by health authorities worldwide; such platforms should induce both arms of the immune system, the humoral and cytotoxic-T-lymphocyte responses. In this study, we have established that a vaccine platform based on the coat protein of papaya mosaic virus (PapMV CP), previously shown to induce a humoral response, can induce major histocompatibility complex (MHC) class I cross-presentation of HLA A*0201 epitopes from gp100, a melanoma antigen, and from influenza virus M1 matrix protein. PapMV proteins were able to assemble into stable virus-like particles (VLPs) in a crystalline and repetitive structure. When we pulsed HLA A*0201+ antigen-presenting cells (APCs) with the recombinant PapMV FLU or gp100, we noted that antigen-specific CD8+ T cells were highly reactive to these APCs, demonstrating that the epitope from the VLPs were processed and loaded on the MHC class I complex. APCs were preincubated with two different proteasome inhibitors, which did not affect the efficiency of peptide presentation on MHC class I. Classical presentation from an endogenous antigen was abolished in the same conditions. Clearly, antigen presentation mediated by the PapMV system was proteasome independent. Finally, PapMV-pulsed APCs had the capacity to expand highly avid antigen-specific T cells against the influenza virus M1 HLA-A*0201 epitope when cocultured with autologous peripheral blood mononuclear cells. This study demonstrates the potential of PapMV for MHC class I cross-presentation and for the expansion of human antigen-specific T cells. It makes VLPs from PapMV CP a very attractive platform to trigger cellular responses for vaccine development against chronic infectious diseases and cancers. PMID- 17121796 TI - Amelioration of influenza virus pathogenesis in chickens attributed to the enhanced interferon-inducing capacity of a virus with a truncated NS1 gene. AB - Avian influenza virus (AIV) A/turkey/Oregon/71-SEPRL (TK/OR/71-SEPRL) (H7N3) encodes a full-length NS1 protein and is a weak inducer of interferon (IFN). A variant, TK/OR/71-delNS1 (H7N3), produces a truncated NS1 protein and is a strong inducer of IFN. These otherwise genetically related variants differ 20-fold in their capacities to induce IFN in primary chicken embryo cells but are similar in their sensitivities to the action of IFN. Furthermore, the weak IFN-inducing strain actively suppresses IFN induction in cells that are otherwise programmed to produce it. These phenotypic differences are attributed to the enhanced IFN inducing capacity that characterizes type A influenza virus strains that produce defective NS1 protein. The pathogenesis of these two variants was evaluated in 1 day-old and 4-week-old chickens. The cell tropisms of both viruses were similar. However, the lesions in chickens produced by the weak IFN inducer were more severe and differed somewhat in character from those observed for the strong IFN inducer. Differences in lesions included the nature of inflammation, the rate of resolution of the infection, and the extent of viral replication and/or virus dissemination. The amelioration of pathogenesis is attributed to the higher levels of IFN produced by the variant encoding the truncated NS1 protein and the antiviral state subsequently induced by that IFN. The high titer of virus observed in kidney tissue ( approximately 10(9) 50% embryo lethal doses/g) from 1 day-old chickens infected intravenously by the weak IFN-inducing strain is attributed to the capacity of chicken kidney cells to activate the hemagglutinin fusion peptide along with their unresponsiveness to inducers of IFN as measured in vitro. Thus, the IFN-inducing capacity of AIV appears to be a significant factor in regulating the pathogenesis, virulence, and viral transmission of AIV in chickens. This suggests that the IFN-inducing and IFN induction suppression phenotypes of AIV should be considered when characterizing strains of influenza virus. PMID- 17121797 TI - Structural and functional characterization of sapovirus RNA-dependent RNA polymerase. AB - Sapoviruses are one of the major agents of acute gastroenteritis in childhood. They form a tight genetic cluster (genus) in the Caliciviridae family that regroups both animal and human pathogenic strains. No permissive tissue culture has been developed for human sapovirus, limiting its characterization to surrogate systems. We report here on the first extensive characterization of the key enzyme of replication, the RNA-dependent RNA polymerase (RdRp) associated with the 3D(pol)-like protein. Enzymatically active sapovirus 3D(pol) and its defective mutant were expressed in Escherichia coli and purified. The overall structure of the sapovirus 3D(pol) was determined by X-ray crystallography to 2.32-A resolution. It revealed a right hand fold typical for template-dependent polynucleotide polymerases. The carboxyl terminus is located within the active site cleft, as observed in the RdRp of some (norovirus) but not other (lagovirus) caliciviruses. Sapovirus 3D(pol) prefers Mn(2+) over Mg(2+) but may utilize either as a cofactor in vitro. In a synthetic RNA template-dependent reaction, sapovirus 3D(pol) synthesizes a double-stranded RNA or labels the template 3' terminus by terminal transferase activity. Initiation of RNA synthesis occurs de novo on heteropolymeric templates or in a primer-dependent manner on polyadenylated templates. Strikingly, this mode of initiation of RNA synthesis was also described for norovirus, but not for lagovirus, suggesting structural and functional homologies in the RNA-dependent RNA polymerase of human pathogenic caliciviruses. This first experimental evidence makes sapovirus 3D(pol) an attractive target for developing drugs to control calicivirus infection in humans. PMID- 17121798 TI - A single-nucleotide synonymous mutation in the gag gene controlling human immunodeficiency virus type 1 virion production. AB - Viral factors as well as host ones play major roles in the disease progression of human immunodeficiency virus type 1 (HIV-1) infection. We have examined cytotoxic T-lymphocyte activity and HIV-1 DNA PCR results of 312 high-risk seronegative drug users in northern Thailand and identified four seronegative cases positive for both assays. Furthermore, we have identified a synonymous mutation in nucleotide position 75 of the gag p17 gene (A426G) of HIV-1 that belongs to the CRF01_AE virus circulating in Thailand. The replication-competent HIV-1 clone containing the A426G mutation demonstrated a dramatic reduction of virion production and perturbation of viral morphogenesis without affecting viral protein synthesis in cells. PMID- 17121799 TI - PTP-H2 and PTP-H3 from Microplitis demolitor Bracovirus localize to focal adhesions and are antiphagocytic in insect immune cells. AB - Viruses in the family Polydnaviridae are symbiotically associated with parasitoid wasps. Wasps inject polydnaviruses (PDVs) when laying an egg into their insect host, and expression of viral gene products causes several physiological alterations, including immunosuppression, that allow the wasp's progeny to develop. As with other PDVs, most Microplitis demolitor bracovirus (MdBV) genes are related variants that form gene families. The largest MdBV gene family includes 13 members that encode predicted proteins related to protein tyrosine phosphatases (PTPs). Sequence analysis during the present study indicated that five PTP family members (PTP-H2, -H3, -N1, and -N2) have fully conserved catalytic domains, whereas other family members exhibited replacements, deletions, or rearrangements of amino acids considered essential for tyrosine phosphatase activity. Expression studies indicated that most MdBV PTP genes are expressed in virus-infected host insects, with transcript abundance usually being highest in hemocytes. MdBV-infected hemocytes also exhibited higher levels of tyrosine phosphatase activity than noninfected hemocytes. We produced expression constructs for four of the most abundantly expressed PTP family members and conducted functional studies with hemocyte-like Drosophila S2 cells. These experiments suggested that recombinant PTP-H2 and PTP-H3 are functional tyrosine phosphatases whereas PTP-H1 and PTP-J1 are not. PTP-H2 and -H3 localized to focal adhesions in S2 cells, and coexpression with another MdBV gene product, Glc1.8, resulted in complete inhibition of phagocytosis. PMID- 17121800 TI - Proinflammatory cytokine gene induction by human T-cell leukemia virus type 1 (HTLV-1) and HTLV-2 Tax in primary human glial cells. AB - Infection with human T-cell leukemia virus type 1 (HTLV-1) can result in the development of HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP), a chronic inflammatory disease of the central nervous system (CNS). HTLV-2 is highly related to HTLV-1 at the genetic level and shares a high degree of sequence homology, but infection with HTLV-2 is relatively nonpathogenic compared to HTLV-1. Although the pathogenesis of HAM/TSP remains to be fully elucidated, previous evidence suggests that elevated levels of the proinflammatory cytokines in the CNS are associated with neuropathogenesis. We demonstrate that HTLV-1 infection in astrogliomas results in a robust induction of interleukin-1beta (IL-1beta), IL-1alpha, tumor necrosis factor alpha (TNF alpha), TNF-beta, and IL-6 expression. HTLV encodes for a viral transcriptional transactivator protein named Tax that also induces the transcription of cellular genes. To investigate and compare the effects of Tax1 and Tax2 expression on the dysregulation of proinflammatory cytokines, lentivirus vectors were used to transduce primary human astrocytomas and oligodendrogliomas. The expression of Tax1 in primary human astrocytomas and oligodendrogliomas resulted in significantly higher levels of proinflammatory cytokine gene expression compared to Tax2. Notably, Tax1 expression uniquely sensitized primary human astrocytomas to apoptosis. A Tax2/Tax1 chimera encoding the C-terminal 53 amino acids of the Tax1 fused to the Tax2 gene (Tax(221)) demonstrated a phenotype that resembled Tax1, with respect to proinflammatory cytokine gene expression and sensitization to apoptosis. The patterns of differential cytokine induction and sensitization to apoptosis displayed by Tax1 and Tax2 may reflect differences relating to the heightened neuropathogenicity associated with HTLV-1 infection and the development of HAM/TSP. PMID- 17121801 TI - Interaction between a 54-kilodalton mammalian cell surface protein and cowpea mosaic virus. AB - Cowpea mosaic virus (CPMV), a plant virus that is a member of the picornavirus superfamily, is increasingly being used for nanotechnology applications, including material science, vascular imaging, vaccine development, and targeted drug delivery. For these applications, it is critical to understand the in vivo interactions of CPMV within the mammalian system. Although the bioavailability of CPMV in the mouse has been demonstrated, the specific interactions between CPMV and mammalian cells need to be characterized further. Here we demonstrate that although the host range for replication of CPMV is confined to plants, mammalian cells nevertheless bind and internalize CPMV in significant amounts. This binding is mediated by a conserved 54-kDa protein found on the plasma membranes of both human and murine cell lines. Studies using a deficient cell line, deglycosidases, and glycosylation inhibitors showed that the CPMV binding protein (CPMV-BP) is not glycosylated. A possible 47-kDa isoform of the CPMV-BP was also detected in the organelle and nuclear subcellular fraction prepared from murine fibroblasts. Further characterization of CPMV-BP is important to understand how CPMV is trafficked through the mammalian system and may shed light on how picornaviruses may have evolved between plant and animal hosts. PMID- 17121802 TI - Comparative analysis of twelve genomes of three novel group 2c and group 2d coronaviruses reveals unique group and subgroup features. AB - Twelve complete genomes of three novel coronaviruses-bat coronavirus HKU4 (bat CoV HKU4), bat-CoV HKU5 (putative group 2c), and bat-CoV HKU9 (putative group 2d) were sequenced. Comparative genome analysis showed that the various open reading frames (ORFs) of the genomes of the three coronaviruses had significantly higher amino acid identities to those of other group 2 coronaviruses than group 1 and 3 coronaviruses. Phylogenetic trees constructed using chymotrypsin-like protease, RNA-dependent RNA polymerase, helicase, spike, and nucleocapsid all showed that the group 2a and 2b and putative group 2c and 2d coronaviruses are more closely related to each other than to group 1 and 3 coronaviruses. Unique genomic features distinguishing between these four subgroups, including the number of papain-like proteases, the presence or absence of hemagglutinin esterase, small ORFs between the membrane and nucleocapsid genes and ORFs (NS7a and NS7b), bulged stem-loop and pseudoknot structures downstream of the nucleocapsid gene, transcription regulatory sequence, and ribosomal recognition signal for the envelope gene, were also observed. This is the first time that NS7a and NS7b downstream of the nucleocapsid gene has been found in a group 2 coronavirus. The high Ka/Ks ratio of NS7a and NS7b in bat-CoV HKU9 implies that these two group 2d specific genes are under high selective pressure and hence are rapidly evolving. The four subgroups of group 2 coronaviruses probably originated from a common ancestor. Further molecular epidemiological studies on coronaviruses in the bats of other countries, as well as in other animals, and complete genome sequencing will shed more light on coronavirus diversity and their evolutionary histories. PMID- 17121803 TI - Silencing of both beta-TrCP1 and HOS (beta-TrCP2) is required to suppress human immunodeficiency virus type 1 Vpu-mediated CD4 down-modulation. AB - The human immunodeficiency virus type 1 (HIV-1) Vpu protein interacts with CD4 within the endoplasmic reticula of infected cells and targets CD4 for degradation through interaction with beta-TrCP1. Mammals possess a homologue of beta-TrCP1, HOS, which is also named beta-TrCP2. We show by coimmunoprecipitation experiments that beta-TrCP2 binds Vpu and is able to induce CD4 down-modulation as efficiently as beta-TrCP1. In two different cell lines, HeLa CD4+ and Jurkat, Vpu mediated CD4 down-modulation could not be reversed through the individual silencing of endogenous beta-TrCP1 or beta-TrCP2 but instead required the two genes to be silenced simultaneously. PMID- 17121804 TI - Rhinovirus activates interleukin-8 expression via a Src/p110beta phosphatidylinositol 3-kinase/Akt pathway in human airway epithelial cells. AB - Rhinovirus (RV) is responsible for the majority of common colds and triggers exacerbations of asthma and chronic obstructive lung disease. We have shown that RV serotype 39 (RV39) infection activates phosphatidylinositol 3 (PI 3)-kinase and the serine threonine kinase Akt minutes after infection and that the activation of PI 3-kinase and Akt is required for maximal interleukin-8 (IL-8) expression. Here, we further examine the contributions of Src and PI 3-kinase activation to RV-induced Akt activation and IL-8 expression. Confocal fluorescent microscopy of 16HBE14o- human bronchial epithelial cells showed rapid (10-min) colocalization of RV39 with Src, p85alpha PI 3-kinase, p110beta PI 3-kinase, Akt and Cit-Akt-PH, a fluorescent Akt pleckstrin homology domain which binds PI(3,4,5)P(3). The chemical Src inhibitor PP2 {4-amino-5-(4-chlorophenyl)-7-(t butyl)pyrazolo [3,4-d]pyrimidine} and the PI 3-kinase inhibitor LY294002 each inhibited Akt phosphorylation and the colocalization of RV39 with Akt. Digoxigenin-tagged RV coprecipitated with a Crosstide kinase likely to be Akt, and inhibition of Src blocked kinase activity. Digoxigenin-tagged RV39 colocalized with the lipid raft marker ceramide. In 16HBE14o- and primary mucociliary differentiated human bronchial epithelial cells, inhibition of Src kinase activity with the Src family chemical inhibitor PP2, dominant-negative Src (K297R), and Src small interfering RNA (siRNA) each inhibited RV39-induced IL-8 expression. siRNA against p110beta PI 3-kinase also inhibited IL-8 expression. These data demonstrate that, in the context of RV infection, Src and p110beta PI 3-kinase are upstream activators of Akt and the IL-8 promoter and that RV colocalizes with Src, PI 3-kinase, and Akt in lipid rafts. PMID- 17121805 TI - E6AP-dependent degradation of DLG4/PSD95 by high-risk human papillomavirus type 18 E6 protein. AB - In most cervical cancers, DNAs of high-risk mucosotropic human papillomaviruses (HPVs), such as types 16 and 18, are maintained so as to express two viral proteins, E6 and E7, suggesting that they play important roles in carcinogenesis. The carboxy-terminal PDZ domain-binding motif of the E6 proteins is in fact essential for transformation of rodent cells and induction of hyperplasia in E6 transgenic mouse skin. To date, seven PDZ domain-containing proteins, including DLG1/hDLG, which is a human homologue of the Drosophila discs large tumor suppressor (Dlg), have been identified as targets of high-risk HPV E6 proteins. Here, we describe DLG4/PSD95, another human homologue of Dlg, as a novel E6 target. DLG4 was found to be expressed in normal human cells, including cervical keratinocytes, but only to a limited extent in both HPV-positive and HPV-negative cervical cancer cell lines. Expression of HPV18 E6 in HCK1T decreased DLG4 levels more strongly than did HPV16 E6, the carboxy-terminal motif of the proteins being critical for binding and degradation of DLG4 in vitro. DLG4 levels were restored by expression of either E6AP-specific short hairpin RNA or bovine papillomavirus type 1 E2 in HeLa but not CaSki or SiHa cells, reflecting downregulation of DLG4 mRNA as opposed to protein by an HPV-independent mechanism in HPV16-positive cancer lines. The tumorigenicity of CaSki cells was strongly inhibited by forced expression of DLG4, while growth in culture was not inhibited at all. These results suggest that DLG4 may function as a tumor suppressor in the development of HPV-associated cancers. PMID- 17121806 TI - Type-specific epitopes targeted by monoclonal antibodies with exceptionally potent neutralizing activities for selected strains of human immunodeficiency virus type 1 map to a common region of the V2 domain of gp120 and differ only at single positions from the clade B consensus sequence. AB - Only a few monoclonal antibodies (MAbs) have been isolated that recognize conserved sites in human immunodeficiency virus type 1 (HIV-1) Env proteins and possess broad neutralizing activities. Other MAbs directed against targets in various domains of Env have been described that are strongly neutralizing, but they possess limited breadth. One such MAb, 2909, possesses a uniquely potent neutralizing activity specific for a quaternary epitope on SF162 Env that requires the presence of both the V2 and the V3 domains. We now show that replacement of the SF162 V3 sequence with consensus V3 sequences of multiple subtypes led to attenuated but still potent neutralization by 2909 and that the main determinants for the type specificity of 2909 reside in the V2 domain. A substitution at position 160 completely eliminated 2909 reactivity, and mutations at position 167 either attenuated or potentiated neutralization by this antibody. Different substitutions at the same positions in V2 were previously shown to introduce epitopes recognized by MAbs 10/76b and C108g and to allow potent neutralization by these MAbs. Two substitutions at key positions in the V2 domain of JR-FL Env also allowed potent expression of the 2909 epitope, and single substitutions in YU2 V2 were sufficient for expression of the 2909, C108g, and 10/76b epitopes. These results demonstrate that the minimal epitopes for 2909, C108g, and 10/76b differed from that of the clade B consensus sequence only at single positions and suggest that all three MAbs recognize distinct variants of a relatively conserved sequence in V2 that is a particularly sensitive mediator of HIV-1 neutralization. PMID- 17121807 TI - Involvement of Hsp90 in assembly and nuclear import of influenza virus RNA polymerase subunits. AB - Transcription and replication of the influenza virus RNA genome occur in the nuclei of infected cells through the viral RNA-dependent RNA polymerase consisting of PB1, PB2, and PA. We previously identified a host factor designated RAF-1 (RNA polymerase activating factor 1) that stimulates viral RNA synthesis. RAF-1 is found to be identical to Hsp90. Here, we examined the intracellular localization of Hsp90 and viral RNA polymerase subunits and their molecular interaction. Hsp90 was found to interact with PB2 and PB1, and it was relocalized to the nucleus upon viral infection. We found that the nuclear transport of Hsp90 occurs in cells expressing PB2 alone. The nuclear transport of Hsp90 was in parallel with that of the viral RNA polymerase binary complexes, either PB1 and PB2 or PB1 and PA, as well as with that of PB2 alone. Hsp90 also interacted with the binary RNA polymerase complex PB1-PB2, and it was dissociated from the PB1 PB2 complex upon its association with PA. Furthermore, Hsp90 could form a stable PB1-PB2-Hsp90 complex prior to the formation of a ternary polymerase complex by the assembly of PA in the infected cells. These results suggest that Hsp90 is involved in the assembly and nuclear transport of viral RNA polymerase subunits, possibly as a molecular chaperone for the polymerase subunits prior to the formation of a mature ternary polymerase complex. PMID- 17121808 TI - IkappaB kinase subunits alpha and gamma are required for activation of NF-kappaB and induction of apoptosis by mammalian reovirus. AB - Reoviruses induce apoptosis both in cultured cells and in vivo. Apoptosis plays a major role in the pathogenesis of reovirus encephalitis and myocarditis in infected mice. Reovirus-induced apoptosis is dependent on the activation of transcription factor NF-kappaB and downstream cellular genes. To better understand the mechanism of NF-kappaB activation by reovirus, NF-kappaB signaling intermediates under reovirus control were investigated at the level of Rel, IkappaB, and IkappaB kinase (IKK) proteins. We found that reovirus infection leads initially to nuclear translocation of p50 and RelA, followed by delayed mobilization of c-Rel and p52. This biphasic pattern of Rel protein activation is associated with the degradation of the NF-kappaB inhibitor IkappaBalpha but not the structurally related inhibitors IkappaBbeta or IkappaBepsilon. Using IKK subunit-specific small interfering RNAs and cells deficient in individual IKK subunits, we demonstrate that IKKalpha but not IKKbeta is required for reovirus induced NF-kappaB activation and apoptosis. Despite the preferential usage of IKKalpha, both NF-kappaB activation and apoptosis were attenuated in cells lacking IKKgamma/Nemo, an essential regulatory subunit of IKKbeta. Moreover, deletion of the gene encoding NF-kappaB-inducing kinase, which is known to modulate IKKalpha function, had no inhibitory effect on either response in reovirus-infected cells. Collectively, these findings indicate a novel pathway of NF-kappaB/Rel activation involving IKKalpha and IKKgamma/Nemo, which together mediate the expression of downstream proapoptotic genes in reovirus-infected cells. PMID- 17121809 TI - Effect of rotavirus strain on the murine model of biliary atresia. AB - Biliary atresia is a devastating disorder of the newborn in which afflicted infants develop inflammation and fibrosis of the extrahepatic biliary tract, resulting in cirrhosis and end-stage liver disease. Infection with a virus is thought to be a contributing factor in the etiology of biliary atresia. In the murine model of biliary atresia, perinatal exposure to rhesus rotavirus (RRV) results in biliary epithelial cell infection causing bile duct obstruction. The purpose of this study was to determine if tropism for the biliary epithelial cell was unique to RRV. Newborn mice underwent intraperitoneal injection with five strains of rotavirus: RRV (simian), SA11-FM (simian/bovine), SA11-SM (simian), EDIM (murine), and Wa (human). RRV and SA11-FM caused clinical manifestations of bile duct obstruction and high mortality. SA11-SM caused clinical signs of hepatobiliary injury but the mortality was markedly reduced. EDIM and Wa caused no sign of hepatobiliary disease. The systemic and temporal distribution of viral protein and live virus varied according to the injected strain. Immunohistochemistry revealed that RRV and SA11-FM targeted the biliary epithelial cells. In contrast, SA11-SM was found in the liver but in not in the biliary epithelium. These results indicate that strain-specific characteristics dictate tropism for cells of hepatobiliary origin which in turn impact the ability to induce the murine model of biliary atresia. PMID- 17121810 TI - beta-Spectrin functions independently of Ankyrin to regulate the establishment and maintenance of axon connections in the Drosophila embryonic CNS. AB - alpha- and beta-Spectrin are major components of a submembrane cytoskeletal network connecting actin filaments to integral plasma membrane proteins. Besides its structural role in red blood cells, the Spectrin network is thought to function in non-erythroid cells during protein targeting and membrane domain formation. Here, we demonstrate that beta-Spectrin is required in neurons for proper midline axon guidance in the Drosophila embryonic CNS. In beta-spectrin mutants many axons inappropriately cross the CNS midline, suggesting a role for beta-Spectrin in midline repulsion. Surprisingly, neither the Ankyrin-binding nor the pleckstrin homology (PH) domains of beta-Spectrin are required for accurate guidance decisions. alpha-Spectrin is dependent upon beta-Spectrin for its normal subcellular localization and/or maintenance, whereas alpha-spectrin mutants exhibit a redistribution of beta-Spectrin to the axon scaffold. beta-spectrin mutants show specific dose-dependent genetic interactions with the midline repellent slit and its neuronal receptor roundabout (robo), but not with other guidance molecules. The results suggest that beta-Spectrin contributes to midline repulsion through the regulation of Slit-Robo pathway components. We propose that the Spectrin network is playing a role independently of Ankyrin in the establishment and/or maintenance of specialized membrane domains containing guidance molecules that ensure the fidelity of axon repulsion at the midline. PMID- 17121811 TI - A functional genomics analysis of the B56 isoforms of Drosophila protein phosphatase 2A. AB - Members of the B56 family of protein phosphatase 2A (PP2A) regulatory subunits play crucial roles in Drosophila cell survival. Distinct functions of two B56 subunits were investigated using a combination of RNA interference, DNA microarrays, and proteomics. RNA interference-mediated knockdown of the B56-1 subunit (PP2A-B') but not the catalytic (mts) or B56-2 subunit (wdb) of PP2A resulted in increased expression of the apoptotic inducers reaper and sickle. Co knockdown of B56-1 with reaper, but not with sickle, reduced the apoptosis caused by depletion of the B56 subunits. Two-dimensional gel electrophoresis and mass spectrometry identified proteins modified in cells depleted of PP2A subunits. These included generation of caspase-dependent cleavage products, increases in protein abundance, and covalent modifications. Results suggested that up regulation of the ribosome-associated protein stubarista can serve as a sensitive marker of apoptosis. Up-regulation of transcripts for multiple glutathione transferases and other proteins suggested that loss of PP2A affected pathways involved in the response to oxidative stress. Knockdown of PP2A elevated basal JNK activity and substantially decreased activation of ERK in response to oxidative stress. The results reveal that the B56-containing isoform of PP2A functions within multiple signaling pathways, including those that regulate expression of reaper and the response to oxidative stress, thus promoting cell survival in Drosophila. PMID- 17121812 TI - CARPs are ubiquitin ligases that promote MDM2-independent p53 and phospho p53ser20 degradation. AB - Caspase 8/10-associated RING proteins (CARPs) are a recently described family of protein ubiquitin ligases that interact with and negatively regulate death receptor-mediated apoptosis. Because CARPs are overexpressed in cancer and their silencing reduces cell viability and sensitizes tumor cells to chemotherapeutic agents, we investigated their relationship to p53 tumor suppressor signaling. p53 is a major determinant of chemosensitivity, and its levels are increased following DNA damage through N-terminal phosphorylation and inhibition of degradation. Although p53 is well known to be negatively regulated by several ubiquitin ligases including MDM2, none are known to target phosphorylated p53 for degradation. CARPs physically interact with and ubiquitinate p53, targeting it for degradation in the absence of MDM2. Serine 20-phosphorylated p53 is also ubiquitinated by CARPs. CARP silencing stimulates p53 expression and promotes downstream effects, including transcriptional activation and tumor suppression. PMID- 17121813 TI - A role for Lte1p (a low temperature essential protein involved in mitosis) in proprotein processing in the yeast secretory pathway. AB - We previously identified six single gene disruptions in Saccharomyces cerevisiae that allow enhanced immunoreactive insulin secretion primarily because of defective Kex2p-mediated endoproteolytic processing. Five eis mutants disrupted established VPS (vacuolar protein sorting) genes, The sixth, LTE1, is a Low Temperature (<15 degrees C) Essential gene encoding a large protein with potential guanine nucleotide exchange (GEF) domains. Lte1p functions as a positive regulator of the mitotic GTPase Tem1p, and overexpression of Tem1p suppresses the low temperature mitotic defect of lte1. By sequence analysis, Tem1p has highest similarity to Vps21p (yeast homolog of mammalian Rab5). Unlike TEM1, LTE1 is not restricted to mitosis but is expressed throughout the cell cycle. Lte1p function in interphase cells is largely unknown. Here we confirm the eis phenotype of lte1 mutant cells and demonstrate a defect in proalpha factor processing that is rescued by expression of full-length Lte1p but not a C terminally truncated Lte1p lacking its GEF homology domain. Neither overexpression of Tem1p nor 13 other structurally related GTPases can suppress the secretory proprotein processing defect. However, overexpression of Vps21p selectively restores proprotein processing in a manner dependent upon the active GTP-bound form of the GTPase. By contrast, a vps21 mutant produces a synthetic defect with lte1 in proprotein processing, as well as a synthetic growth defect. Together, the data underscore a link between the mitotic regulator, Lte1p, and protein processing and trafficking in the secretory/endosomal system. PMID- 17121814 TI - Pannexin-1 couples to maitotoxin- and nigericin-induced interleukin-1beta release through a dye uptake-independent pathway. AB - Pannexin-1 is a recently identified membrane protein that can act as a nonselective pore permeable to dyes such as ethidium when ectopically expressed. Blockade of pannexin-1 in macrophage endogenously expressing the ATP-gated P2X7 receptor (P2X7R) blocks the initial dye uptake, but not the ionic current, and also blocks processing and release of interleukin-1beta (IL-1beta) in response to P2X7R activation. These results suggest that pannexin-1 may be a hemichannel activated by the P2X7R to provide the conduit for dye uptake and downstream signaling to processing and release of IL-1beta. We have pursued this hypothesis by measuring dye uptake and IL-1beta processing and release in mouse J774 macrophage in response to P2X7R activation and to maitotoxin and nigericin, two agents considered to evoke IL-1beta release via the same mechanism. The experiments were carried out over time periods during which no lactate dehydrogenase was released from cells to examine only noncytolytic pathways. P2X7R activation evoked dye uptake that could be separated into two components by pannexin-1 inhibition: an initial rapid phase and a slower pannexin-1-independent phase. Maitotoxin-evoked dye uptake was unaltered by pannexin-1 inhibition. Nigericin did not induce dye uptake. Inhibition of pannexin-1 blocked caspase-1 and IL-1beta processing and release in response to all three stimuli. Thus, although pannexin-1 is required for IL-1beta release in response to maitotoxin, nigericin, and ATP, a mechanism distinct from pannexin-1 hemichannel activation must underlie the former two processes. PMID- 17121815 TI - Filamin-regulated F-actin assembly is essential for morphogenesis and controls phototaxis in Dictyostelium. AB - Dictyostelium strains lacking the F-actin cross-linking protein filamin (ddFLN) have a severe phototaxis defect at the multicellular slug stage. Filamins are rod shaped homodimers that cross-link the actin cytoskeleton into highly viscous, orthogonal networks. Each monomer chain of filamin is comprised of an F-actin binding domain and a rod domain. In rescue experiments only intact filamin re established correct phototaxis in filamin minus mutants, whereas C-terminally truncated filamin proteins that had lost the dimerization domain and molecules lacking internal repeats but retaining the dimerization domain did not rescue the phototaxis defect. Deletion of individual rod repeats also changed their subcellular localization, and mutant filamins in general were less enriched at the cell cortex as compared with the full-length protein and were increasingly present in the cytoplasm. For correct phototaxis ddFLN is only required at the tip of the slug because expression under control of the cell type-specific extracellular-matrix protein A (ecmA) promoter and mixing experiments with wild type cells supported phototactic orientation. Likewise, in chimeric slugs wild type cells were primarily found at the tip of the slug, which acts as an organizer in Dictyostelium morphogenesis. PMID- 17121816 TI - The structure of a cyanobacterial bicarbonate transport protein, CmpA. AB - Cyanobacteria, blue-green algae, are the most abundant autotrophs in aquatic environments and form the base of the food chain by fixing carbon and nitrogen into cellular biomass. To compensate for the low selectivity of Rubisco for CO2 over O2, cyanobacteria have developed highly efficient CO2-concentrating machinery of which the ABC transport system CmpABCD from Synechocystis PCC 6803 is one component. Here, we have described the structure of the bicarbonate binding protein CmpA in the absence and presence of bicarbonate and carbonic acid. CmpA is highly homologous to the nitrate transport protein NrtA. CmpA binds carbonic acid at the entrance to the ligand-binding pocket, whereas bicarbonate binds in nearly an identical location compared with nitrate binding to NrtA. Unexpectedly, bicarbonate binding is accompanied by a metal ion, identified as Ca2+ via inductively coupled plasma optical emission spectrometry. The binding of bicarbonate and metal appears to be highly cooperative and suggests that CmpA may co-transport bicarbonate and calcium or that calcium acts a cofactor in bicarbonate transport. PMID- 17121817 TI - Yersinia protein kinase YopO is activated by a novel G-actin binding process. AB - Pathogenic bacteria of the genus Yersinia employ a type III secretion system to inject effector proteins (Yops) into host cells. The Yops down-regulate host cell functions through unique biochemical activities. YopO, a serine/threonine kinase required for Yersinia virulence, is activated by host cell actin via an unknown process. Here we show that YopO kinase is activated by formation of a 1:1 complex with monomeric (G) actin but is unresponsive to filamentous (F) actin. Two separate G-actin binding sites, one in the N-terminal kinase region (amino acids 89-440) and one in the C-terminal guanine nucleotide dissociation inhibitor-like region (amino acids 441-729) of YopO, were identified. Actin binding to both of these sites was necessary for effective autophosphorylation of YopO on amino acids Ser-90 and Ser-95. A S90A/S95A YopO mutant was strongly reduced in substrate phosphorylation, suggesting that autophosphorylation activates YopO kinase activity. In cells the kinase activity of YopO regulated rounding/arborization and was specifically required for inhibition of Yersinia YadA-dependent phagocytosis. Thus, YopO kinase is activated by a novel G-actin binding process, and this appears to be crucial for its anti-host cell functions. PMID- 17121818 TI - Chemical determinants involved in anandamide-induced inhibition of T-type calcium channels. AB - Anandamide, originally described as an endocannabinoid, is the main representative molecule of a new class of signaling lipids including endocannabinoids and N-acyl-related molecules, eicosanoids, and fatty acids. Bioactive lipids regulate neuronal excitability by acting on G-protein-coupled receptors (such as CB1) but also directly modulate various ionic conductances including voltage-activated T-type calcium channels (T-channels). However, little is known about the properties and the specificity of this new class of molecules on their various targets. In this study, we have investigated the chemical determinants involved in anandamide-induced inhibition of the three cloned T channels: Ca(V)3.1, Ca(V)3.2, and Ca(V)3.3. We show that both the hydroxyl group and the alkyl chain of anandamide are key determinants of its effects on T currents. As follows, T-currents are also inhibited by fatty acids. Inhibition of the three Ca(V)3 currents by anandamide and arachidonic acid does not involve enzymatic metabolism and occurs in cell-free inside-out patches. Inhibition of T currents by fatty acids and N-acyl ethanolamides depends on the degree of unsaturation but not on the alkyl chain length and consequently is not restricted to eicosanoids. Inhibition increases for polyunsaturated fatty acids comprising 18-22 carbons when cis-double bonds are close to the carboxyl group. Therefore the major natural (food-supplied) and mammalian endogenous fatty acids including gamma-linolenic acid, mead acid, and arachidonic acid as well as the fully polyunsaturated omega3-fatty acids that are enriched in fish oil eicosapentaenoic and docosahexaenoic acids are potent inhibitors of T-currents, which possibly contribute to their physiological functions. PMID- 17121819 TI - Soluble androgen receptor oligomers underlie pathology in a mouse model of spinobulbar muscular atrophy. AB - In polyglutamine diseases such as X-linked spinobulbar muscular atrophy (SBMA), it is unknown whether the toxic form of the protein is an insoluble or soluble aggregate or a monomer. We have addressed this question by studying a full-length androgen receptor (AR) mouse model of SBMA. We used biochemistry and atomic force microscopy to immunopurify oligomers soluble after ultracentrifugation that are comprised of a single approximately 50-kDa N-terminal polyglutamine-containing AR fragment. AR oligomers appeared several weeks prior to symptom onset, were distinct and temporally dissociated from intranuclear inclusions, and disappeared rapidly after castration, which halts disease. This is the first demonstration of soluble AR oligomers in vivo and suggests that they underlie neurodegeneration in SBMA. PMID- 17121820 TI - Direct demonstration of the flexibility of the glycosylated proline-threonine linker in the Cellulomonas fimi Xylanase Cex through NMR spectroscopic analysis. AB - The modular xylanase Cex (or CfXyn10A) from Cellulomonas fimi consists of an N terminal catalytic domain and a C-terminal cellulose-binding domain, joined by a glycosylated proline-threonine (PT) linker. To characterize the conformation and dynamics of the Cex linker and the consequences of its modification, we have used NMR spectroscopy to study full-length Cex in its nonglycosylated ( approximately 47 kDa) and glycosylated ( approximately 51 kDa) forms. The PT linker lacks any predominant structure in either form as indicated by random coil amide chemical shifts. Furthermore, heteronuclear (1)H-(15)N nuclear Overhauser effect relaxation measurements demonstrate that the linker is flexible on the ns-to-ps time scale and that glycosylation partially dampens this flexibility. The catalytic and cellulose-binding domains also exhibit identical amide chemical shifts whether in isolation or in the context of either unmodified or glycosylated full-length Cex. Therefore, there are no noncovalent interactions between the two domains of Cex or between either domain and the linker. This conclusion is supported by the distinct (15)N relaxation properties of the two domains, as well as their differential alignment within a magnetic field by Pf1 phage particles. These data demonstrate that the PT linker is a flexible tether, joining the structurally independent catalytic and cellulose-binding domains of Cex in an ensemble of conformations; however, more extended forms may predominate because of restrictions imparted by the alternating proline residues. This supports the postulate that the binding-domain anchors Cex to the surface of cellulose, whereas the linker provides flexibility for the catalytic domain to hydrolyze nearby hemicellulose (xylan) chains. PMID- 17121821 TI - The C-terminal products of cellular prion protein processing, C1 and C2, exert distinct influence on p53-dependent staurosporine-induced caspase-3 activation. AB - The cellular prion protein (PrP(c)) undergoes various endopro-teolytic attacks within its N-terminal domain, leading to the production of C-terminal fragments (C) tethered to the plasma membrane and soluble N-terminal peptides (N). One of these cleavages occurs at position 110/111, thereby generating C1 and N1 products. We have reported that disintegrins ADAM-10, -9, and -17 participate either directly or indirectly to this proteolytic event. An alternative proteolytic event taking place around residue 90 yields C2 and N2 fragments. The putative function of these proteolytic fragments remained to be established. We have set up two novel human embryonic kidney 293 cell lines stably overexpressing either C1 or C2. We show that C1 potentiates staurosporine-induced caspase-3 activation through a p53-dependent mechanism. Thus, C1 positively controls p53 transcription and mRNA levels and increases p53-like immunoreactivity and activity. C1-induced caspase-3 activation remained unaffected by the blockade of endocytosis in HEK 293 cells and was abolished in p53-deficient fibroblasts. Conversely, overexpression of the C2 fragment did not significantly sensitize HEK 293 cells to apoptotic stimuli and did not modify p53 mRNA levels or activity. Therefore, the nature of the proteolytic cleavage taking place on PrP(c) yielded C-terminal catabolites with distinct function and could be seen as a switch mechanism controlling the function of the PrP(c) in cell survival. PMID- 17121822 TI - Inefficient proofreading and biased error rates during inaccurate DNA synthesis by a mutant derivative of Saccharomyces cerevisiae DNA polymerase delta. AB - DNA polymerase delta (pol delta) is a high fidelity eukaryotic enzyme that participates in DNA repair and is essential for DNA replication. Toward the goal of dissecting its multiple biological functions, here we describe the biochemical properties of Saccharomyces cerevisiae pol delta with a methionine replacing conserved leucine 612 at the polymerase active site. Compared with wild type pol delta, L612M pol delta has normal processivity and slightly higher polymerase specific activity. L612M pol delta also has normal 3' exonuclease activity, yet it is impaired in partitioning mismatches to the exonuclease active site, thereby reducing DNA synthesis fidelity. Error rates in vitro for L612M pol delta are elevated for both base substitutions and single base deletions but in a highly biased manner. For each of the six possible pairs of reciprocal mismatches that could arise during replication of complementary DNA strands to account for any particular base substitution in vivo (e.g. T-dGMP or A-dCMP for T to C transitions), L612M pol delta error rates are substantially higher for one mismatch than the other. These results provide a biochemical explanation for our observation, which confirms earlier genetic studies, that a haploid pol3-L612M S. cerevisiae strain has an elevated spontaneous mutation rate that is likely due to reduced replication fidelity in vivo. PMID- 17121823 TI - Structural characterization of the active site of the PduO-type ATP:Co(I)rrinoid adenosyltransferase from Lactobacillus reuteri. AB - The three-dimensional crystal structure of the PduO-type corrinoid adenosyltransferase from Lactobacillus reuteri (LrPduO) has been solved to 1.68-A resolution. The functional assignment of LrPduO as a corrinoid adenosyltransferase was confirmed by in vivo and in vitro evidence. The enzyme has an apparent Km(ATP) of 2.2 microM and Km(Cobalamin) of 0.13 microM and a kcat of 0.025 s(-1). Co-crystallization of the enzyme with Mg-ATP resulted in well defined electron density for an N-terminal loop that had been disordered in other PduO-type enzyme structures. This newly defined N-terminal loop makes up the lower portion of the enzyme active site with the other half being contributed from an adjacent subunit. These results provide the first detailed description of the enzyme active site for a PduO-type adenosyltransferase and identify a unique ATP binding motif at the protein N terminus. The molecular architecture at the active site offers valuable new insight into the role of various residues responsible for the human disease methylmalonic aciduria. PMID- 17121824 TI - H(2)O(2)-induced kinetic and chemical modifications of smooth muscle myosin: correlation to effects of H(2)O(2) on airway smooth muscle. AB - The effect of H(2)O(2) on smooth muscle heavy meromyosin (HMM) and subfragment 1 (S1) was examined. The number of molecules that retained the ability to bind ATP and the actinactivated rate of P(i) release were measured by single-turnover kinetics. H(2)O(2) treatment caused a decrease in HMM regulation from 800- to 27 fold. For unphosphorylated and phosphorylated heavy meromyosin and for S1, approximately 50% of the molecules lost the ability to bind to ATP. H(2)O(2) treatment in the presence of EDTA protected against ATPase inactivation and against the loss of total ATP binding. Inactivation of S1 versus time correlated to a loss of reactive thiols. Treatment of H(2)O(2)-inactivated phosphorylated HMM or S1 with dithiothreitol partially reactivated the ATPase but had no effect on total ATP binding. H(2)O(2)-inactivated S1 contained a prominent cross-link between the N-terminal 65-kDa and C-terminal 26-kDa heavy chain regions. Mass spectral studies revealed that at least seven thiols in the heavy chain and the essential light chain were oxidized to cysteic acid. In thiophosphorylated porcine tracheal muscle strips at pCa 9 + 2.1 mM ATP, H(2)O(2) caused a approximately 50% decrease in the amplitude but did not alter the rate of force generation, suggesting that H(2)O(2) directly affects the force generating complex. Dithiothreitol treatment reversed the H(2)O(2) inhibition of the maximal force by approximately 50%. These data, when compared with the in vitro kinetic data, are consistent with a H(2)O(2)-induced loss of functional myosin heads in the muscle. PMID- 17121825 TI - Inverted factor access and slow reversion characterize SWI/SNF-altered nucleosome dimers. AB - Human SWI/SNF (hSWI/SNF) is an ATP-dependent chromatin remodeling complex with important functions in activation and repression of cellular genes. Previously, we showed that hSWI/SNF creates structurally altered dimers from mononucleosome cores. More recently we found that hSWI/SNF also generates abundant structurally altered dinucleosomes, called altosomes, on polynucleosomal templates. Here, we find that dimers revert to normal nucleosomes at a similar rate as altosomes and can also be cleaved to yield nucleosomal particles with mobilities similar to mononucleosomes. Using these and other shared properties we propose a single model for both types of hSWI/SNF product. In addition, we further characterize the accessibility of altered dimers to transcription factors, and find that the DNA in dimers is most accessible in the middle and least accessible at the ends, directly opposite the profile of normal mononucleosomes. We also find that transcription factor binding can influence the ratio of normal nucleosomes and dimers as hSWI/SNF products. Implications for the interplay between hSWI/SNF products and transcription factors are discussed. PMID- 17121826 TI - Molecular determinants of substrate selectivity of a novel organic cation transporter (PMAT) in the SLC29 family. AB - Plasma membrane monoamine transporter (PMAT or ENT4) is a newly cloned transporter assigned to the equilibrative nucleoside transporter (ENT) family (SLC29). Unlike ENT1-3, PMAT mainly functions as a polyspecific organic cation transporter. In this study, we investigated the molecular mechanisms underlying the unique substrate selectivity of PMAT. By constructing chimeras between human PMAT and ENT1, we showed that a chimera consisting of transmembrane domains (TM) 1-6 of PMAT and TM7-11 of hENT1 behaved like PMAT, transporting 1-methyl-4 phenylpyridinium (MPP+, an organic cation) but not uridine (a nucleoside), suggesting that TM1-6 contains critical domains responsible for substrate recognition. To identify residues important for the cation selectivity of PMAT, 10 negatively charged residues were chosen and substituted with alanine. Five of the alanine mutants retained PMAT activity, and four were non-functional due to impaired targeting to the plasma membrane. However, alanine substitution at Glu(206) in TM5 abolished PMAT activity without affecting cell surface expression. Eliminating the charge at Glu(206) (E206Q) resulted in loss of organic cation transport activity, whereas conserving the negative charge (E206D) restored transporter function. Interestingly, mutant E206Q, which possesses the equivalent residue in ENT1, gained uridine transport activity. Thr(220), another residue in TM5, also showed an effect on PMAT activity. Helical wheel analysis of TM5 revealed a distinct amphipathic pattern with Glu(206) and Thr(220) clustered in the center of the hydrophilic face. In summary, our results suggest that Glu(206) functions as a critical charge sensor for cationic substrates and TM5 forms part of the substrate permeation pathway in PMAT. PMID- 17121827 TI - Specific high affinity interactions of monomeric endotoxin.protein complexes with Toll-like receptor 4 ectodomain. AB - Potent Toll-like receptor 4 (TLR4) activation by endotoxin has been intensely studied, but the molecular requirements for endotoxin interaction with TLR4 are still incompletely defined. Ligand-receptor interactions involving endotoxin and TLR4 were characterized using monomeric endotoxin.protein complexes of high specific radioactivity. The binding of endotoxin.MD-2 to the TLR4 ectodomain (TLR4ECD) and transfer of endotoxin from CD14 to MD-2/TLR4ECD were demonstrated using HEK293T-conditioned medium containing TLR4ECD+/-MD-2. These interactions are specific, of high affinity (KD<300 pm), and consistent with the molecular requirements for potent cell activation by endotoxin. Both reactions result in the formation of a Mr approximately 190,000 complex composed of endotoxin, MD-2, and TLR4ECD. CD14 facilitates transfer of endotoxin to MD-2 (TLR4) but is not a stable component of the endotoxin.MD-2/TLR4 complex. The ability to assay specific high affinity interactions of monomeric endotoxin.protein complexes with TLR4ECD should allow better definition of the structural requirements for endotoxin-induced TLR4 activation. PMID- 17121828 TI - An antisense transcript induced by Wnt/beta-catenin signaling decreases E2F4. AB - Wnt signaling induces the nuclear accumulation of beta-catenin and transcription of specific target genes via the DNA-binding proteins TCF/Lef. Although all known beta-catenin target genes encode proteins, genome-wide RNA profiling studies indicate that many transcripts do not have this capability. Transcription factor binding sites associated with these noncoding transcripts can be identified using unbiased techniques such as serial analysis of chromatin occupancy (SACO). We used this method to identify a beta-catenin-regulated antisense RNA expressed in HCT116 colorectal carcinoma cells, a cellular model of activated beta-catenin signaling. Genomic signature tags designating putative beta-catenin-binding sites mapped to the 3'-untranslated region (3'-UTR) of the E2F4 gene. We showed that both beta-catenin and TCF4 bind to the E2F4 3'-UTR site in vivo, inducing expression of an E2F4 antisense transcript. LiCl, which mimics Wnt signaling, also induced expression of the E2F4 antisense transcript and decreased E2F4 protein levels. This effect was blocked by a cDNA expressing the E2F4 3'-UTR sense strand. The antisense-mediated decrease in E2F4 protein was reflected by reduced E2F4 association with specific target genes, including CCNA2, CDC2, PCNA, and Rad54. We propose that Wnt/beta-catenin signaling may contribute to colorectal carcinogenesis by reducing the level of the E2F4 cell cycle repressor via an antisense mechanism. PMID- 17121829 TI - Visualization of aggregation of the Rnq1 prion domain and cross-seeding interactions with Sup35NM. AB - Factors triggering the de novo appearance of prions are still poorly understood. In yeast, the appearance of one prion, [PSI(+)], is enhanced by the presence of another prion, [PIN(+)]. The [PSI(+)] and [PIN(+)] prion-forming proteins are, respectively, the translational termination factor Sup35 and the yet poorly characterized Rnq1 protein that is rich in glutamines and asparagines. The prion domain of Rnq1 (RnqPD) polymerizes more readily in vitro than the full-length protein. As is typical for amyloidogenic proteins, the reaction begins with a lag phase, followed by exponential growth. Seeding with pre-formed aggregates significantly shortens the lag. A generic antibody against pre-amyloid oligomer inhibits the unseeded but not the self-seeded reaction. As revealed by electron microscopy, RnqPD polymerizes predominantly into spherical species that eventually agglomerate. We observed infrequent fiber-like structures in samples taken at 4 h of polymerization, but in overnight samples SDS treatment was required to reveal fibers among agglomerates. Polymerization reactions in which RnqPD and the prion domain of Sup35 (Sup35NM) cross-seed each other proceeded with a shortened lag that only depends weakly on the protein concentration. Cross seeded Sup35NM fibers appear to sprout from globular RnqPD aggregates as seen by electron microscopy. RnqPD spherical aggregates appear to associate with and, later occlude, Sup35NM seed fibers. Our kinetic and morphological analyses suggest that, upon cross-seeding, the aggregate provides the surface on which oligomers of the heterologous protein nucleate their subsequent amyloid formation. PMID- 17121830 TI - Differential helical orientations among related G protein-coupled receptors provide a novel mechanism for selectivity. Studies with salvinorin A and the kappa-opioid receptor. AB - Salvinorin A, the active component of the hallucinogenic sage Salvia divinorum, is an apparently selective and highly potent kappa-opioid receptor (KOR) agonist. Salvinorin A is unique among ligands for peptidergic G protein-coupled receptors in being nonnitrogenous and lipid-like in character. To examine the molecular basis for the subtype-selective binding of salvinorin A, we utilized an integrated approach using chimeric opioid receptors, site-directed mutagenesis, the substituted cysteine accessibility method, and molecular modeling and dynamics studies. We discovered that helix 2 is required for salvinorin A binding to KOR and that two residues (Val-108(2.53) and Val-118(2.63)) confer subtype selectivity. Intriguingly, molecular modeling studies predicted that these loci exhibit an indirect effect on salvinorin A binding, presumably through rotation of helix 2. Significantly, and in agreement with our in silico predictions, substituted cysteine accessibility method analysis of helix 2 comparing KOR and the delta-opioid receptor, which has negligible affinity for salvinorin A, revealed that residues known to be important for salvinorin A binding exhibit a differential pattern of water accessibility. These findings imply that differences in the helical orientation of helix 2 are critical for the selectivity of salvinorin A binding to KOR and provide a structurally novel basis for ligand selectivity. PMID- 17121831 TI - Respiratory distress and neonatal lethality in mice lacking Golgi alpha1,2 mannosidase IB involved in N-glycan maturation. AB - There are three mammalian Golgi alpha1,2-mannosidases, encoded by different genes, that form Man5GlcNAc2 from Man(8-9)GlcNAc2 for the biosynthesis of hybrid and complex N-glycans. Northern blot analysis and in situ hybridization indicate that the three paralogs display distinct developmental and tissue-specific expression. The physiological role of Golgi alpha1,2-mannosidase IB was investigated by targeted gene ablation. The null mice have normal gross appearance at birth, but they display respiratory distress and die within a few hours. Histology of fetal lungs the day before birth indicate some delay in development, whereas neonatal lungs show extensive pulmonary hemorrhage in the alveolar region. No significant histopathological changes occur in other tissues. No remarkable ultrastructural differences are detected between wild type and null lungs. The membranes of a subset of bronchiolar epithelial cells are stained with lectins from Phaseolus vulgaris (leukoagglutinin and erythroagglutinin) and Datura stramonium in wild type lungs, but this staining disappears in lungs from null mice. Mass spectrometry of N-glycans from different tissues shows no significant changes in global N-glycans of null mice. Therefore, only a few glycoproteins required for normal lung function depend on alpha1,2-mannosidase IB for maturation. There are no apparent differences in the expression of several lung epithelial cell and endothelial cell markers between null and wild type mice. The alpha1,2-mannosidase IB null phenotype differs from phenotypes caused by ablation of other enzymes in N-glycan biosynthesis and from other mouse gene disruptions that affect pulmonary development and function. PMID- 17121832 TI - Distinctive T cell-suppressive signals from nuclearized type 1 sphingosine 1 phosphate G protein-coupled receptors. AB - Sphingosine 1-phosphate (S1P) generated by cells of innate immunity and the type 1 S1P G protein-coupled receptor (S1P(1)) on mobile T cells constitute a major system for control of lymphoid organ traffic and tissue migration of T cells. Now we show that T cell activation mediated by the T cell antigen receptor translocates plasma membrane S1P(1) to nuclear envelope membranes for association there with G(i/o), Erk (1/2), and other proteins that plasma membrane S1P(1) uses to signal T cell proliferation. However, nuclear S1P(1) and plasma membrane S1P(1) transduce opposite effects of S1P on T cell proliferation and relevant signaling as exemplified by respective decreases and increases in T cell nuclear concentrations of both phospho-Erk and active (phosphorylated) c-Jun. T cell antigen receptor-mediated activation of T cells therefore both eliminates migration responses to S1P by down-regulation of plasma membrane S1P(1) and translocates the S1P-S1P(1) axis into the nuclear domain where signals are directed to transcriptional control of immune functions other than migration. PMID- 17121833 TI - The macrophage cell surface glyceraldehyde-3-phosphate dehydrogenase is a novel transferrin receptor. AB - The reticuloendothelial system plays a major role in iron metabolism. Despite this, the manner in which macrophages handle iron remains poorly understood. Mammalian cells utilize transferrin-dependent mechanisms to acquire iron via transferrin receptors 1 and 2 (TfR1 and TfR2) by receptor-mediated endocytosis. Here, we show for the first time that the glycolytic enzyme glyceraldehyde-3 phosphate dehydrogenase (GAPDH) is localized on human and murine macrophage cell surface. The expression of this surface GAPDH is regulated by the availability of iron in the medium. We further demonstrate that this GAPDH interacts with transferrin and the GAPDH-transferrin complex is subsequently internalized into the early endosomes. Our work sheds new light on the mechanisms involved in regulation of iron, vital for controlling numerous diseases and maintaining normal immune function. Thus, we propose an entirely new avenue for investigation with respect to transferrin uptake and regulation mechanisms in macrophages. PMID- 17121834 TI - Identification of a novel mitochondrial complex containing mitofusin 2 and stomatin-like protein 2. AB - A reverse genetics approach was utilized to discover new proteins that interact with the mitochondrial fusion mediator mitofusin 2 (Mfn2) and that may participate in mitochondrial fusion. In particular, in vivo formaldehyde cross linking of whole HeLa cells and immunoprecipitation with purified Mfn2 antibodies of SDS cell lysates were used to detect an approximately 42-kDa protein. This protein was identified by liquid chromatography and tandem mass spectrometry as stomatin-like protein 2 (Stoml2), previously described as a peripheral plasma membrane protein of unknown function associated with the cytoskeleton of erythrocytes (Wang, Y., and Morrow, J. S. (2000) J. Biol. Chem. 275, 8062-8071). Immunoblot analysis with anti-Stoml2 antibodies showed that Stoml2 could be immunoprecipitated specifically with Mfn2 antibody either from formaldehyde-cross linked and SDS-lysed cells or from cells lysed with digitonin. Subsequent immunocytochemistry and cell fractionation experiments fully supported the conclusion that Stoml2 is indeed a mitochondrial protein. Furthermore, demonstration of mitochondrial membrane potential-dependent import of Stoml2 accompanied by proteolytic processing, together with the results of sublocalization experiments, suggested that Stoml2 is associated with the inner mitochondrial membrane and faces the intermembrane space. Notably, formaldehyde cross-linking revealed a "ladder" of high molecular weight protein species, indicating the presence of high molecular weight Stoml2-Mfn2 hetero-oligomers. Knockdown of Stoml2 by the short interfering RNA approach showed a reduction of the mitochondrial membrane potential, without, however, any obvious changes in mitochondrial morphology. PMID- 17121835 TI - Crystal structures of native and inactivated cis-3-chloroacrylic acid dehalogenase. Structural basis for substrate specificity and inactivation by (R) oxirane-2-carboxylate. AB - The bacterial degradation pathways for the nematocide 1,3-dichloropropene rely on hydrolytic dehalogenation reactions catalyzed by cis- and trans-3-chloroacrylic acid dehalogenases (cis-CaaD and CaaD, respectively). X-ray crystal structures of native cis-CaaD and cis-CaaD inactivated by (R)-oxirane-2-carboxylate were elucidated. They locate four known catalytic residues (Pro-1, Arg-70, Arg-73, and Glu-114) and two previously unknown, potential catalytic residues (His-28 and Tyr 103'). The Y103F and H28A mutants of these latter two residues displayed reductions in cis-CaaD activity confirming their importance in catalysis. The structure of the inactivated enzyme shows covalent modification of the Pro-1 nitrogen atom by (R)-2-hydroxypropanoate at the C3 position. The interactions in the complex implicate Arg-70 or a water molecule bound to Arg-70 as the proton donor for the epoxide ring-opening reaction and Arg-73 and His-28 as primary binding contacts for the carboxylate group. This proposed binding mode places the (R)-enantiomer, but not the (S)-enantiomer, in position to covalently modify Pro 1. The absence of His-28 (or an equivalent) in CaaD could account for the fact that CaaD is not inactivated by either enantiomer. The cis-CaaD structures support a mechanism in which Glu-114 and Tyr-103' activate a water molecule for addition to C3 of the substrate and His-28, Arg-70, and Arg-73 interact with the C1 carboxylate group to assist in substrate binding and polarization. Pro-1 provides a proton at C2. The involvement of His-28 and Tyr-103' distinguishes the cis-CaaD mechanism from the otherwise parallel CaaD mechanism. The two mechanisms probably evolved independently as the result of an early gene duplication of a common ancestor. PMID- 17121836 TI - Cancer cell cycle modulated by a functional coupling between sigma-1 receptors and Cl- channels. AB - The sigma-1 receptor is an intracellular protein characterized as a tumor biomarker whose function remains mysterious. We demonstrate herein for the first time that highly selective sigma ligands inhibit volume-regulated chloride channels (VRCC) in small cell lung cancer and T-leukemia cells. Sigma ligands and VRCC blockers provoked a cell cycle arrest underlined by p27 accumulation. In stably sigma-1 receptor-transfected HEK cells, the proliferation rate was significantly lowered by sigma ligands when compared with control cells. Sigma ligands produced a strong inhibition of VRCC in HEK-transfected cells but not in control HEK. Surprisingly, the activation rate of VRCC was dramatically delayed in HEK-transfected cells in the absence of ligands, indicating that sigma-1 receptors per se modulate cell regulating volume processes in physiological conditions. Volume measurements in hypotonic conditions revealed indeed that the regulatory volume decrease was delayed in HEK-transfected cells and virtually abolished in the presence of igmesine in both HEK-transfected and T-leukemic cells. Moreover, HEK-transfected cells showed a significant resistance to staurosporine-induced apoptosis volume decrease, indicating that sigma-1 receptors protect cancer cells from apoptosis. Altogether, our results show for the first time that sigma-1 receptors modulate "cell destiny" through VRCC and cell volume regulation. PMID- 17121837 TI - Role of ABCG1 and ABCA1 in regulation of neuronal cholesterol efflux to apolipoprotein E discs and suppression of amyloid-beta peptide generation. AB - Maintenance of an adequate supply of cholesterol is important for neuronal function, whereas excess cholesterol promotes amyloid precursor protein (APP) cleavage generating toxic amyloid-beta (Abeta) peptides. To gain insights into the pathways that regulate neuronal cholesterol level, we investigated the potential for reconstituted apolipoprotein E (apoE) discs, resembling nascent lipoprotein complexes in the central nervous system, to stimulate neuronal [3H]cholesterol efflux. ApoE discs potently accelerated cholesterol efflux from primary human neurons and cell lines. The process was saturable (17.5 microg of apoE/ml) and was not influenced by APOE genotype. High performance liquid chromatography analysis of cholesterol and cholesterol metabolites effluxed from neurons indicated that <25% of the released cholesterol was modified to polar products (e.g. 24-hydroxycholesterol) that diffuse from neuronal membranes. Thus, most cholesterol (approximately 75%) appeared to be effluxed from neurons in a native state via a transporter pathway. ATP-binding cassette transporters ABCA1, ABCA2, and ABCG1 were detected in neurons and neuroblastoma cell lines and expression of these cDNAs revealed that ABCA1 and ABCG1 stimulated cholesterol efflux to apoE discs. In addition, ABCA1 and ABCG1 expression in Chinese hamster ovary cells that stably express human APP significantly reduced Abeta generation, whereas ABCA2 did not modulate either cholesterol efflux or Abeta generation. These data indicate that ABCA1 and ABCG1 play a significant role in the regulation of neuronal cholesterol efflux to apoE discs and in suppression of APP processing to generate Abeta peptides. PMID- 17121838 TI - Food intake regulates oleoylethanolamide formation and degradation in the proximal small intestine. AB - Oleoylethanolamide (OEA) is a lipid mediator that inhibits food intake by activating the nuclear receptor peroxisome proliferator-activated receptor-alpha. In the rodent small intestine OEA levels decrease during food deprivation and increase upon refeeding, suggesting that endogenous OEA may participate in the regulation of satiety. Here we show that feeding stimulates OEA mobilization in the mucosal layer of rat duodenum and jejunum but not in the serosal layer from the same intestinal segments in other sections of the gastrointestinal tract (stomach, ileum, colon) or in a broad series of internal organs and tissues (e.g. liver, brain, heart, plasma). Feeding also increases the levels of other unsaturated fatty acid ethanolamides (FAEs) (e.g. linoleoylethanolamide) without affecting those of saturated FAEs (e.g. palmitoylethanolamide). Feeding-induced OEA mobilization is accompanied by enhanced accumulation of OEA-generating N acylphosphatidylethanolamines (NAPEs) increased activity and expression of the OEA-synthesizing enzyme NAPE-phospholipase D, and decreased activity and expression of the OEAdegrading enzyme fatty acid amide hydrolase. Immunostaining studies revealed that NAPE-phospholipase D and fatty acid amide hydrolase are expressed in intestinal enterocytes and lamina propria cells. Collectively, these results indicate that nutrient availability controls OEA mobilization in the mucosa of the proximal intestine through a concerted regulation of OEA biosynthesis and degradation. PMID- 17121839 TI - Nitric oxide down-regulates polo-like kinase 1 through a proximal promoter cell cycle gene homology region. AB - Polo-like kinase 1 (PLK1) is an evolutionarily conserved serine/threonine kinase essential for cell mitosis. As a master cell cycle regulator, p21/Waf1 plays a critical role in cell cycle progression. Nitric oxide (NO.) has been shown to down-regulate PLK1 and up-regulate p21/Waf1 independent of cGMP. Here, the respective roles of p38 MAPK and p21/Waf1 in NO.-mediated PLK1 repression were investigated using differentiated U937 cells that lack soluble guanylate cyclase. NO. was shown to down-regulate both PLK1 mRNA and protein. Nuclear run-on assays and mRNA stability studies demonstrated that the effect of NO. on PLK1 expression was associated with decreased transcription without changes in transcript stability. SB202190, a p38 MAPK inhibitor, prevented transcriptional repression of PLK1 by NO.. Transfection with dominant-negative p38 MAPK mutant eliminated the NO. effect on both p21/Waf1 and PLK1 gene expression. Knockdown of p21/Waf1 with siRNA also substantially reduced the regulatory effect of NO. on PLK1. Reporter gene experiments showed that NO. decreased activity of the PLK1 proximal promoter, an effect that was blocked by p38 MAPK inhibitor. Deletion or mutation of the CDE/CHR promoter site, an element regulated by p21/Waf1, increased base line promoter activity and abolished NO. repression of the PLK1 promoter. Likewise, electrophoretic mobility shift assays with CDE/CHR probe revealed a NO. mediated change in protein-probe complex formation. Competition with various unlabeled CDE/CHR mutant sequences showed that NO. increased nuclear protein binding to intact CHR. These results demonstrate that a NO.-p38 MAPK-p21/Waf1 signal transduction pathway represses PLK1 through a canonical CDE/CHR promoter element. PMID- 17121840 TI - APOBEC3F can inhibit the accumulation of HIV-1 reverse transcription products in the absence of hypermutation. Comparisons with APOBEC3G. AB - APOBEC3F (apolipoprotein B mRNA-editing enzyme catalytic polypeptide 1-like protein 3F) is a cytidine deaminase that, like APOBEC3G, is able to restrict the replication of HIV-1/delta vif. Initial studies revealed high numbers of mutations in the cDNA of viruses produced in the presence of these proteins, suggesting that cytidine deamination underpinned the inhibition of infection. However, we have recently shown that catalytically inactive APOBEC3G proteins, derived through mutation of the C-terminal cytidine deaminase motif, still exert a substantial antiviral effect. Here, we have generated a panel of APOBEC3F mutant proteins and show that the C-terminal cytidine deaminase motif is essential for catalytic activity and that catalytic activity is not necessary for the antiviral effect of APOBEC3F. Furthermore, we demonstrate that the antiviral activities of wild-type and catalytically inactive APOBEC3F and APOBEC3G proteins correspond well with reductions in the accumulation of viral reverse transcription products. Additional comparisons between APOBEC3F and APOBEC3G suggest that the loss of deaminase activity is more detrimental to APOBEC3G function than to APOBEC3F function, as reflected by perturbations to the suppression of reverse transcript accumulation as well as antiviral activity. Taken together, these data suggest that both APOBEC3F and APOBEC3G are able to function as antiviral factors in the absence of cytidine deamination, that this editing-independent activity is an important aspect of APOBEC protein-mediated antiviral phenotypes, but that APOBEC3F may be a better model in which to study it. PMID- 17121841 TI - Genetic evidence for adenylyl cyclase 1 as a target for preventing neuronal excitotoxicity mediated by N-methyl-D-aspartate receptors. AB - The excessive activation of N-methyl-D-aspartate (NMDA) receptors by glutamate results in neuronal excitotoxicity. cAMP is a key second messenger and contributes to NMDA receptor-dependent synaptic plasticity. Adenylyl cyclases 1 (AC1) and 8 (AC8) are the two major calcium-stimulated ACs in the central nervous system. Previous studies demonstrate AC1 and AC8 play important roles in synaptic plasticity, memory, and persistent pain. However, little is known about the possible roles of these two ACs in glutamate-induced neuronal excitotoxicity. Here, we report that genetic deletion of AC1 significantly attenuated neuronal death induced by glutamate in primary cultures of cortical neurons, whereas AC8 deletion did not produce a significant effect. AC1, but not AC8, contributes to intracellular cAMP production following NMDA receptor activation by glutamate in cultured cortical neurons. AC1 is involved in the dynamic modulation of cAMP response element-binding protein activity in neuronal excitotoxicity. To explore the possible roles of AC1 in cell death in vivo, we studied neuronal excitotoxicity induced by an intracortical injection of NMDA. Cortical lesions induced by NMDA were significantly reduced in AC1 but not in AC8 knock-out mice. Our findings provide direct evidence that AC1 plays an important role in neuronal excitotoxicity and may serve as a therapeutic target for preventing excitotoxicity in stroke and neurodegenerative diseases. PMID- 17121842 TI - Regulation of the yeast TSA1 peroxiredoxin by ZAP1 is an adaptive response to the oxidative stress of zinc deficiency. AB - Zinc deficiency is a potential risk factor for disease in humans because it leads to increased oxidative stress and DNA damage. We show here that the yeast Saccharomyces cerevisiae also experiences oxidative stress when zinc-deficient, and we have identified one mechanism yeast cells use to defend themselves against this stress. The Zap1p transcription factor is a central player in the response of yeast to zinc deficiency. To identify genes important for growth in low zinc, DNA microarrays were used to identify genes directly regulated by Zap1p. We found that the TSA1 gene is one such Zap1p target whose expression is increased under zinc deficiency. TSA1 encodes a cytosolic thioredoxin-dependent peroxidase responsible for degrading hydrogen peroxide and organic hydroperoxides. Consistent with its regulation by Zap1p, we showed that tsa1delta mutants have a growth defect in low zinc that can be suppressed by zinc but not by other metals. Anaerobic conditions also suppressed the tsa1delta low zinc growth defect indicating that oxidative stress is the likely cause of the poor growth. Consistent with this hypothesis, we demonstrated that zinc deficiency causes increased reactive oxygen species in wild type cells and that this increase is further exacerbated in tsa1delta mutants. The role of this regulation by Zap1p in limiting oxidative stress in low zinc was confirmed when the Zap1p-binding site was specifically mutated in the chromosomal TSA1 promoter. Thus, we conclude that TSA1 induction by Zap1p is an adaptive response to deal with the increased oxidative stress caused by zinc deficiency. PMID- 17121843 TI - Phosphorylation of glutamate receptor interacting protein 1 regulates surface expression of glutamate receptors. AB - The number of synaptic alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-type glutamate receptors (AMPARs) controls the strength of excitatory transmission. AMPARs cycle between internal endosomal compartments and the plasma membrane. Interactions between the AMPAR subunit GluR2, glutamate receptor interacting protein 1 (GRIP1), and the endosomal protein NEEP21 are essential for correct GluR2 recycling. Here we show that an about 85-kDa protein kinase phosphorylates GRIP1 on serine 917. This kinase is present in NEEP21 immunocomplexes and is activated in okadaic acid-treated neurons. Pulldown assays and atomic force microscopy indicate that phosphorylated GRIP shows reduced binding to NEEP21. AMPA or N-methyl-D-aspartate stimulation of hippocampal neurons induces delayed phosphorylation of the same serine 917. A wild type carboxy-terminal GRIP1 fragment expressed in hippocampal neurons interferes with GluR2 surface expression. On the contrary, a S917D mutant fragment does not interfere with GluR2 surface expression. Likewise, coexpression of GluR2 together with full-length wild type GRIP1 enhances GluR2 surface expression in fibroblasts, whereas full-length GRIP1-S917D had no effect. This indicates that this serine residue is implicated in AMPAR cycling. Our results identify an important regulatory mechanism in the trafficking of AMPAR subunits between internal compartments and the plasma membrane. PMID- 17121844 TI - N-linked oligosaccharides on the low density lipoprotein receptor homolog SorLA/LR11 are modified with terminal GalNAc-4-SO4 in kidney and brain. AB - Sorting protein-related receptor (SorLA/LR11) is a highly conserved mosaic receptor that is expressed by cells in a number of different tissues including principal cells of the collecting ducts in the kidney and neurons in the central and peripheral nervous systems. SorLA/LR11 has features that indicate it serves as a sorting receptor shuttling between the plasma membrane, endosomes, and the Golgi. We have found that a fraction of SorLA/LR11 that is synthesized in the kidney and the brain bears N-linked oligosaccharides that are modified with terminal beta1,4-linked GalNAc-4-SO(4). Oligosaccharides located in the vacuolar sorting (Vps) 10p domain (Vps10p domain) are modified with beta1,4-linked GalNAc when the Vps10p domain is expressed in cells along with either of two recently cloned protein-specific beta1,4GalNAc-transferases, GalNAcTIII and GalNAcTIV. Either of two sequences with basic amino acids located within the Vps10p domain is able to mediate recognition by these beta1,4GalNAc-transferases. The highly specific modification of oligosaccharides in the Vps10p domain of SorLA/LR11 with terminal GalNAc-4-SO(4) suggests that this unusual modification may modulate the interaction of SorLA/LR11 with proteins and influence their trafficking. PMID- 17121845 TI - Sustained JNK activation in response to tumor necrosis factor is mediated by caspases in a cell type-specific manner. AB - In most cell types, tumor necrosis factor (TNF) induces a transient activation of the JNK pathway. However, in NFkappaB-inhibited cells, TNF stimulates also a second sustained phase of JNK activation, which has been implicated in cell death induction. In the present study, we have analyzed the relationship of cell death induction, caspase activity, JNK, and NFkappaB stimulation in the context of TNF signaling in four different cellular systems. In all cases, NFkappaB inhibition enhanced TNF-induced cell death and primed most, but not all, cells for sustained JNK activation. The caspase inhibitor Benzyloxycarbonyl-Val-Ala-Asp(OMe) fluoromethyl ketone (Z-VAD-fmk) and overexpression of the antiapoptotic proteins FLIP-L and Bcl2 differentially blocked transient and sustained JNK activation in NFkappaB-inhibited KB and HaCaT cells, indicating that the two phases of TNF induced JNK activation occur at least in these cellular models by different pathways. Although the broad range caspase inhibitor Z-VAD-fmk and the antioxidant butylated hydroxyanisole interfered with TNF-induced cell death to a varying extent in a cell type-specific manner, inhibition of JNK signaling had no or only a very moderate effect. Notably, the JNK inhibitory effect of neither Z VAD-fmk nor butylated hydroxyanisole was strictly correlated with the capability of these compounds to rescue cells from TNF-induced cell death. Thus, sustained JNK activation by TNF has no obligate role in TNF-induced cell death and is mediated by caspases and reactive oxygen species in a cell type-specific manner. PMID- 17121846 TI - Impact of membrane fusion and proteolysis on SpoIIQ dynamics and interaction with SpoIIIAH. AB - The onset of engulfment-dependent gene expression during Bacillus subtilis sporulation requires the forespore membrane protein SpoIIQ, which recruits mother cell proteins involved in late gene expression to the outer forespore membrane. Engulfment activates the late forespore transcription factor sigmaG, which produces high levels of the secreted SpoIVB protease that is required for activation of the late mother cell transcription factor sigmaK. Engulfment also triggers the proteolytic cleavage of SpoIIQ, an event that depends on the SpoIVB protease but not on sigmaG activity. To determine if SpoIVB directly cleaves SpoIIQ and to determine if this event participates in the onset of late gene expression, we purified SpoIVB, SpoIIQ, and SpoIVFA (another SpoIVB substrate). SpoIVB directly cleaved SpoIIQ at the same site in vitro and in vivo and cleaved SpoIVFA in at least three different locations. SpoIIQ cleavage depends on membrane fusion, but not on sigmaG activity, suggesting that the ability of SpoIVB to cleave substrates is regulated by membrane fusion. We isolated SpoIVB resistant SpoIIQ proteins by random mutagenesis of codons at the cleavage site and demonstrated that SpoIIQ processing is dispensable for spore formation and for activation of late forespore and mother cell gene expression. Fluorescence recovery after photobleaching analysis demonstrated that membrane fusion releases SpoIIQ from an immobile complex, an event that could allow SpoIVB to cleave SpoIIQ. We propose that this membrane fusion-dependent reorganization in the complex, rather than SpoIIQ proteolysis itself, is necessary for the onset of late transcription. PMID- 17121847 TI - Cooperation of DEF6 with activated Rac in regulating cell morphology. AB - Rho-family GTPases have been implicated in actin remodeling and subsequent morphologic changes in various cells. DEF6, a pleckstrin homology domain containing protein, has been reported to regulate Rho-family GTPases as a guanine nucleotide exchange factor. Here, we demonstrate that DEF6 also has the property of cooperating with activated Rac1. DEF6 bound selectively to Rac1 loaded with GTP. The interaction is mediated by the effector domain of Rac1. Overexpression of GFP-DEF6 together with constitutively active Rac1 in COS-7 cells significantly changed their cell shape; this was not seen in the absence of activated Rac1. This effect of DEF6 on cellular morphology was shown to be independent of its guanine nucleotide exchange activity. Because DEF6 does not contain any sequences previously known to interact with Rac, we explored the domain necessary for the binding. The amino-terminal portion and central parts of DEF6 were required for the binding. Finally, we succeeded in creating mutants of DEF6 with point mutations in the amino-terminal portion, which abrogate the binding to activated Rac1. These mutants did not exhibit the morphologic change in COS-7 cells when they were co-expressed with activated Rac1. These results suggest that DEF6 not only activates Rho-family GTPases but also cooperates with activated Rac1 to exert its cellular function. PMID- 17121848 TI - UBPY-mediated epidermal growth factor receptor (EGFR) de-ubiquitination promotes EGFR degradation. AB - Whereas poly-ubiquitination targets protein substrates for proteasomal degradation, mono-ubiquitination is known to regulate protein trafficking in the endosomal system and to target cargo proteins for lysosomal degradation. The role of the de-ubiquitinating enzymes AMSH and UBPY in endosomal trafficking of cargo proteins such as the epidermal growth factor receptor (EGFR) has only very recently been the subject of study and is already a matter of debate. Although one report (Mizuno, E., Iura, T., Mukai, A., Yoshimori, T., Kitamura, N., and Komada, M. (2005) Mol. Biol. Cell 16, 5163-5174) concludes that UBPY negatively regulates EGFR degradation by de-ubiquitinating the EGFR on endosomes, another report (Row, P. E., Prior, I. A., McCullough, J., Clague, M. J., and Urbe, S. (2006) J. Biol. Chem. 281, 12618-12624) concludes that UBPY-mediated EGFR de ubiquitination is essential for EGFR degradation. Here, we demonstrate that Usp8/UBPY, the mammalian ortholog of budding yeast Ubp4/Doa4, constitutively co precipitates in a bivalent manner with the EGFR. Moreover, UBPY is a substrate for Src-family tyrosine kinases that are activated after ligand-induced EGFR activation. Using overexpression of three different recombinant dominant negative UBPY mutants (UBPY C748A mutant, UBPY 1-505, and UBPY 640-1080) in NIH3T3 and HEK293 cells, we demonstrate that UBPY affects both constitutive and ligand induced (i) EGFR ubiquitination, (ii) EGFR expression levels, and (iii) the appearance of intermediate EGFR degradation products as well as (iv) downstream mitogen-activated protein kinase signal transduction. Our findings provide further evidence in favor of the model that UBPY-mediated EGFR de-ubiquitination promotes EGFR degradation. PMID- 17121849 TI - Calpain is required for the rapid, calcium-dependent repair of wounded plasma membrane. AB - Mammalian cells require extracellular calcium ion to undergo rapid plasma membrane repair seconds after mechanical damage. Utilizing transformed fibroblasts from calpain small subunit knock-out (Capns1-/-) mouse embryos, we now show that the heterodimeric, typical subclass of calpains is required for calcium-mediated survival after plasma membrane damage caused by scraping a cell monolayer. Survival of scrape-damaged Capns1-/- cells was unaffected by calcium in the scraping medium, whereas more Capns1+/+ cells survived when calcium was present. Calcium-mediated survival was increased when Capns1-/- cells were scraped in the presence of purified m- or mu-calpain. Survival rates of scraped Capns1+/+, HFL-1, or Chinese hamster ovary cells were decreased by the calpain inhibitor, calpeptin, or the highly specific calpain inhibitor protein, calpastatin. Capns1-/- cells failed to reseal following laser-induced membrane disruption, demonstrating that their decreased survival after scraping resulted, at least in part, from failed membrane repair. Proteomic and immunologic analyses demonstrated that the known calpain substrates talin and vimentin were exposed at the cell surface and processed by calpain following cell scraping. Autoproteolytic activation of calpain at the scrape site was evident at the earliest time point analyzed and appeared to precede proteolysis of talin and vimentin. The results indicate that conventional calpains are required for calcium-facilitated survival after plasma membrane damage and may act by localized remodeling of the cortical cytoskeleton at the injury site. PMID- 17121850 TI - In vivo and in vitro degradation of heparan sulfate (HS) proteoglycans by HPR1 in pancreatic adenocarcinomas. Loss of cell surface HS suppresses fibroblast growth factor 2-mediated cell signaling and proliferation. AB - Heparan sulfate proteoglycans (HSPGs) function as a co-receptor for heparin binding growth factors, such as fibroblast growth factors (FGFs) and heparin bound epidermal growth factor (HB-EGF). The HS side chain of HSPGs can be cleaved by HPR1 (heparanase-1), an endoglycosidase that is overexpressed in many types of malignancies. In the present study, we demonstrated that HPR1 expression in pancreatic adenocarcinomas inversely correlated with the presence of heparan sulfate (HS) in the basement membrane. In vitro cell culture study revealed that cell surface HS levels inversely correlated with HPR1 activity in five pancreatic cancer cell lysates and their conditioned media. Heparin and PI-88, two HPR1 inhibitors, were able to increase cell surface HS levels in PANC-1 cells in a dose-dependent manner. The ability of HPR1 to degrade cell surface HS was confirmed by showing that cell surface HS levels were increased in HT1080 cells stably transfected with the HPR1 antisense gene but was decreased in the cells overexpressing HPR1. Further studies showed that PI-88 and heparin were able to stimulate PANC-1 cell proliferation in the absence or presence of exogenous FGF2, whereas exogenous HPR1 was able to inhibit PANC-1 cell proliferation in a dose dependent manner. Modulation of PANC-1 cell proliferation by HPR1 or HPR1 inhibitors corresponded with the inhibition or activation of the mitogen activated protein kinase. Our results suggest that HPR1 expressed in pancreatic adenocarcinomas can suppress the proliferation of pancreatic tumor cells in response to the growth factors that require HSPGs as their co-receptors. PMID- 17121851 TI - Stress-induced c-Jun-dependent Vitamin D receptor (VDR) activation dissects the non-classical VDR pathway from the classical VDR activity. AB - Vitamin D receptor (VDR) is a ligand-dependent transcription factor that mediates vitamin D(3)-induced gene expression. Our previous work has established that stress MAPK signaling stimulates VDR expression (Qi, X., Pramank, R., Wang, J., Schultz, R. M., Maitra, R. K., Han, J., DeLuca, H. F., and Chen, G. (2002) J. Biol. Chem. 277, 25884-25892) and VDR inhibits cell death in response to p38 MAPK activation (Qi, X., Tang, J., Pramanik, R., Schultz, R. M., Shirasawa, S., Sasazuki, T., Han, J., and Chen, G. (2004) J. Biol. Chem. 279, 22138-22144). Here we show that c-Jun is essential for VDR expression and VDR in turn inhibits c-Jun dependent cell death by non-classical mechanisms. In response to stress c-Jun is recruited to the Vdr promoter before VDR protein expression is induced. The necessary and sufficient role of c-Jun in VDR expression was established by the fact that c-Jun knock-out decreases VDR expression, whereas c-Jun restoration recovers its activity. Existence of the non-classical VDR pathway was suggested by a requirement of both c-Jun and VDR in stress-induced VDR activity and further demonstrated by VDR inhibiting c-Jun-dependent cell death independent of its classical transcriptional activity and independent of vitamin D(3). c-Jun is also required for vitamin D(3)-induced classical VDR transcriptional activity by a mechanism likely involving physical interactions between c-Jun and VDR proteins. These results together reveal a non-classical mechanism by which VDR acts as a c Jun/AP-1 target gene to modify c-Jun activity in stress response through increased protein expression independent of classical transcriptional regulations. PMID- 17121852 TI - RBCK1, a protein kinase CbetaI (PKCbetaI)-interacting protein, regulates PKCbeta dependent function. AB - RBCK1 (RBCC protein interacting with PKC 1) has originally been identified as a protein kinase CbetaI (PKCbetaI)-binding partner by a two-hybrid screen and as one of the gene transcripts that increases during adult cardiac hypertrophy. To address whether RBCK1 and PKCbetaI functions are interconnected, we used cultured neonatal myocytes where we previously found that the activity of PKCbetaI is required for an increase in cell size, also called hypertrophy. In this study, we showed that acute treatment of cardiac myocytes with phenylephrine, a prohypertrophic stimulant, transiently increased the association of RBCK1 with PKCbetaI within 1 min. A prolonged phenylephrine treatment also resulted in an increase of the interaction of the two proteins. Endogenous RBCK1 protein levels increased upon phenylephrine-induced hypertrophy. Further, adenovirus-based RBCK1 overexpression in the absence of phenylephrine increased cardiac cell size. This RBCK1-mediated hypertrophy required PKCbeta activity, since the increase in cell size was inhibited when the RBCK1-expressing cells were treated with PKCbeta selective antagonists, supporting our previous observation that both PKCbetaI and PKCbetaII are required for hypertrophy. Unexpectedly, RBCK1-induced increased cell size was inhibited by phenylephrine. This effect correlated with a decrease in the level of both PKCbeta isoforms. Most importantly, RNA interference for RBCK1 significantly inhibited the increase in cell size of cardiac myocytes following phenylephrine treatment. Our results suggest that RBCK1 binds PKCbetaI and is a key regulator of PKCbetaI function in cells and that, together with PKCbetaII, the three proteins are essential for developmental hypertrophy of cardiac myocytes. PMID- 17121853 TI - Understanding a transcriptional paradigm at the molecular level. The structure of yeast Gal80p. AB - In yeast, the GAL genes encode the enzymes required for normal galactose metabolism. Regulation of these genes in response to the organism being challenged with galactose has served as a paradigm for eukaryotic transcriptional control over the last 50 years. Three proteins, the activator Gal4p, the repressor Gal80p, and the ligand sensor Gal3p, control the switch between inert and active gene expression. Gal80p, the focus of this investigation, plays a pivotal role both in terms of repressing the activity of Gal4p and allowing the GAL switch to respond to galactose. Here we present the three-dimensional structure of Gal80p from Kluyveromyces lactis and show that it is structurally homologous to glucose-fructose oxidoreductase, an enzyme in the sorbitol gluconate pathway. Our results clearly define the overall tertiary and quaternary structure of Gal80p and suggest that Gal4p and Gal3p bind to Gal80p at distinct but overlapping sites. In addition to providing a molecular basis for previous biochemical and genetic studies, our structure demonstrates that much of the enzymatic scaffold of the oxidoreductase has been maintained in Gal80p, but it is utilized in a very different manner to facilitate transcriptional regulation. PMID- 17121854 TI - Essential roles for Fe65, Alzheimer amyloid precursor-binding protein, in the cellular response to DNA damage. AB - Fe65 interacts with the cytosolic domain of the Alzheimer amyloid precursor protein (APP). The functions of the Fe65 are still unknown. To address this point we generated Fe65 knockout (KO) mice. These mice do not show any obvious phenotype; however, when fibroblasts (mouse embryonic fibroblasts), isolated from Fe65 KO embryos, were exposed to low doses of DNA damaging agents, such as etoposide or H2O2, an increased sensitivity to genotoxic stress, compared with wild type animals, clearly emerged. Accordingly, brain extracts from Fe65 KO mice, exposed to non-lethal doses of ionizing radiations, showed high levels of gamma-H2AX and p53, thus demonstrating a higher sensitivity to X-rays than wild type mice. Nuclear Fe65 is necessary to rescue the observed phenotype, and few minutes after the exposure of MEFs to DNA damaging agents, Fe65 undergoes phosphorylation in the nucleus. With a similar timing, the proteolytic processing of APP is rapidly affected by the genotoxic stress: in fact, the cleavage of the APP COOH-terminal fragments by gamma-secretase is induced soon after the exposure of cells to etoposide, in a Fe65-dependent manner. These results demonstrate that Fe65 plays an essential role in the response of the cells to DNA damage. PMID- 17121855 TI - Suppression of calpain-dependent cleavage of the CDK5 activator p35 to p25 by site-specific phosphorylation. AB - Cdk5 is a proline-directed Ser/Thr protein kinase predominantly expressed in postmitotic neurons together with its activator, p35. N-terminal truncation of p35 to p25 by calpain results in deregulation of Cdk5 and contributes to neuronal cell death associated with several neurodegenerative diseases. Previously we reported that p35 occurred as a phosphoprotein, phospho-p35 levels changed with neuronal maturation, and that phosphorylation of p35 affected its vulnerability to calpain cleavage. Here, we identify the p35 residues Ser(8) and Thr(138) as the major sites of phosphorylation by Cdk5. Mutagenesis of these sites to unphosphorylatable Ala increased susceptibility to calpain in cultured cells and neurons while changing them to phosphomimetic glutamate-attenuated cleavage. Furthermore, phosphorylation state-specific antibodies to these sites revealed that Thr(138) was dephosphorylated in adult rat, although both Ser(8) and Thr(138) were phosphorylated in prenatal brains. In cultured neurons, inhibition of protein phosphatases converted phosho-Ser(8) p35 to dual phospho Ser(8)/Thr(138) p35 and conferred resistance to calpain cleavage. These results suggest phosphorylation of Thr(138) predominantly defines the susceptibility of p35 to calpain-dependent cleavage and that dephosphorylation of this site is a critical determinant of Cdk5-p25-induced cell death associated with neurodegeneration. PMID- 17121856 TI - Site-specific acetylation of p53 directs selective transcription complex assembly. AB - Histone deacetylase (HDAC) inhibitors are being investigated as possible adjuvant therapies for a number of diseases, including cancer. In addition to stabilization of acetylated histones, HDAC inhibitors stabilize the acetylation of a number of transcription factors, including p53. This study investigates the action of two HDAC inhibitors, CG-1521 and trichostatin A, which stabilize Ac-Lys 373 p53 and Ac-Lys-382 p53, respectively, in LNCaP prostate cancer cells. Real time PCR demonstrates that CG-1521 induces p21 transcription whereas trichostatin A does not alter the steady state level of p21 mRNA. Co-immunoprecipitation demonstrates that the selective acetylation of p53 directs the recruitment of mutually exclusive coactivator complexes on the p53 response elements in the p21 promoter. Furthermore, the co-activator complexes initiate the recruitment of the components of the basal transcription apparatus to the basal promoter with markedly different outcomes because only Ac-Lys-373 p53 promotes the assembly of the basal transcriptional apparatus on the p21 promoter. These data highlight the profound effects of post-translational modification, including acetylation, on the function of p53. The data also suggest a novel and critically important role for protein acetylation/deacetylation in the assembly of active transcription processes that may be as important as classical phosphorylation/dephosphorylation. PMID- 17121857 TI - Hydrodynamic studies on the quaternary structure of recombinant mouse Purbeta. AB - Purbeta is a gene regulatory factor belonging to a family of highly conserved nucleic acid-binding proteins related by their ability to preferentially bind single-stranded DNA or RNA sequences rich in purine nucleotides. In conjunction with Puralpha, Purbeta has been implicated in transcriptional and translational repression of genes encoding contractile proteins found in the heart and vasculature. Although several models of sequence-specific DNA recognition, strand separation, and activator inhibition by oligomeric Puralpha and Purbeta have been proposed, it is currently unclear whether protein-protein interaction is a prerequisite to, or a consequence of nucleic acid binding. In this study, a recombinant protein purification scheme was devised to yield homogenous mouse Purbeta devoid of nucleic acid. Recombinant Purbeta was then subjected to light scattering and analytical ultracentrifugation analyses to assess the size, shape, and oligomeric state of the purified protein in solution. Results of laser light scattering and sedimentation velocity experiments indicated that Purbeta reversibly self-associates in the absence of nucleic acid. Both approaches independently showed that the hydrodynamic shape of the Purbeta homodimer is markedly asymmetric and non-spherical. Sedimentation velocity analyses indicated that dimeric Purbeta has a sedimentation coefficient of 3.96 Svedberg, a frictional coefficient ratio (f/f(0)) of 1.60, and a hydrodynamic radius of 4.43 nm. These values were consistent with those determined by independent dynamic light scattering studies. Sedimentation equilibrium analyses confirmed that Purbeta self-associates in a reversible monomer-dimer equilibrium characterized by a K(d) = 1.13 +/- 0.27 microm. PMID- 17121858 TI - Highly conserved residues Asp-197 and His-250 in Agp1 phytochrome control the proton affinity of the chromophore and Pfr formation. AB - The mutants H250A and D197A of Agp1 phytochrome from Agrobacterium tumefaciens were prepared and investigated by different spectroscopic and biochemical methods. Asp-197 and His-250 are highly conserved amino acids and are part of the hydrogen-bonding network that involves the chromophore. Both substitutions cause a destabilization of the protonated chromophore in the Pr state as revealed by resonance Raman and UV-visible absorption spectroscopy. Titration experiments demonstrate a lowering of the pK(a) from 11.1 (wild type) to 8.8 in H250A and 7.2 in D197A. Photoconversion of the mutants does not lead to the Pfr state. H250A is arrested in a meta-Rc-like state in which the chromophore is deprotonated. For H250A and the wild-type protein, deprotonation of the chromophore in meta-Rc is coupled to the release of a proton to the external medium, whereas the subsequent proton re-uptake, linked to the formation of the Pfr state in the wild-type protein, is not observed for H250A. No transient proton exchange with the external medium occurs in D197A, suggesting that Asp-197 may be the proton release group. Both mutants do not undergo the photo-induced protein structural changes that in the wild-type protein are detectable by size exclusion chromatography. These conformational changes are, therefore, attributed to the meta-Rc --> Pfr transition and most likely coupled to the transient proton re uptake. The present results demonstrate that Asp-197 and His-250 are essential for stabilizing the protonated chromophore structure in the parent Pr state, which is required for the primary photochemical process, and for the complete photo-induced conversion to the Pfr state. PMID- 17121859 TI - Reversible sequestration of active site cysteines in a 2Fe-2S-bridged dimer provides a mechanism for glutaredoxin 2 regulation in human mitochondria. AB - Human mitochondrial glutaredoxin 2 (GLRX2), which controls intracellular redox balance and apoptosis, exists in a dynamic equilibrium of enzymatically active monomers and quiescent dimers. Crystal structures of both monomeric and dimeric forms of human GLRX2 reveal a distinct glutathione binding mode and show a 2Fe-2S bridged dimer. The iron-sulfur cluster is coordinated through the N-terminal active site cysteine, Cys-37, and reduced glutathione. The structures indicate that the enzyme can be inhibited by a high GSH/GSSG ratio either by forming a 2Fe 2S-bridged dimer that locks away the N-terminal active site cysteine or by binding non-covalently and blocking the active site as seen in the monomer. The properties that permit GLRX2, and not other glutaredoxins, to form an iron-sulfur containing dimer are likely due to the proline-to-serine substitution in the active site motif, allowing the main chain more flexibility in this area and providing polar interaction with the stabilizing glutathione. This appears to be a novel use of an iron-sulfur cluster in which binding of the cluster inactivates the protein by sequestering active site residues and where loss of the cluster through changes in subcellular redox status creates a catalytically active protein. Under oxidizing conditions, the dimers would readily separate into iron free active monomers, providing a structural explanation for glutaredoxin activation under oxidative stress. PMID- 17121860 TI - Biochemical and functional analysis of the assembly of full-length Sup35p and its prion-forming domain. AB - The protein Sup35 has prion properties. Its aggregation is at the origin of the [PSI(+)] trait in Saccharomyces cerevisiae. In vitro, the N-terminal domain of Sup35p alone or with the middle domain assembles into fibrils that exhibit the characteristics of amyloids. The vast majority of in vitro studies on the assembly of Sup35p have been performed using Sup35pNM, as fibrils made of Sup35pNM assembled in vitro propagate [PSI(+)] when reintroduced into yeast cells. Little is known about the assembly of full-length Sup35p and the role of the functional C-terminal domain of the protein. Here we report a systematic comparison of the biochemical and assembly properties of full-length Sup35p and Sup35pNM. We show that the native structure of the C-terminal domain is retained within the fibrils. We determined the size of Sup35p nuclei and the critical concentration for assembly that both differ from that of Sup35pNM. We demonstrate that Sup35pNM co-assembles with the full-length protein and that fibrils made of Sup35p or Sup35pNM seed the assembly of soluble Sup35pNM and Sup35p with different efficiencies. Finally, we show that fibrils made of full-length Sup35p induce with higher efficiency [PSI(+)] appearance as compared with those made of Sup35pNM. Our findings reveal differences and similarities in the assembly of Sup35p and its NM fragment and validate the use of Sup35pNM in studying some aspects of Sup35p aggregation but also underline the importance of using full length Sup35p in studying prion propagation both in vivo and in vitro. PMID- 17121861 TI - Processing of 3'-phosphoglycolate-terminated DNA double strand breaks by Artemis nuclease. AB - The Artemis nuclease is required for V(D)J recombination and for repair of an as yet undefined subset of radiation-induced DNA double strand breaks. To assess the possibility that Artemis acts on oxidatively modified double strand break termini, its activity toward model DNA substrates, bearing either 3'-hydroxyl or 3'-phosphoglycolate moieties, was examined. A 3'-phosphoglycolate had little effect on Artemis-mediated trimming of long 3' overhangs (> or =9 nucleotides), which were efficiently trimmed to 4-5 nucleotides. However, 3'-phosphoglycolates on overhangs of 4-5 bases promoted Artemis-mediated removal of a single 3' terminal nucleotide, while at least 2 nucleotides were trimmed from identical hydroxyl-terminated substrates. Artemis also efficiently removed a single nucleotide from a phosphoglycolate-terminated 3-base 3' overhang, while leaving an analogous hydroxyl-terminated overhang largely intact. Such removal was completely dependent on DNA-dependent protein kinase and ATP and was largely dependent on Ku, which markedly stimulated Artemis activity toward all 3' overhangs. Together, these data suggest that efficient Artemis-mediated cleavage of 3' overhangs requires a minimum of 2 nucleotides, or a nucleotide plus a phosphoglycolate, 3' to the cleavage site, as well as 2 unpaired nucleotides 5' to the cleavage site. Shorter 3'-phosphoglycolate-terminated overhangs and blunt ends were also processed by Artemis but much more slowly. Consistent with a role for Artemis in repair of terminally blocked double strand breaks in vivo, human cells lacking Artemis exhibited hypersensitivity to x-rays, bleomycin, and neocarzinostatin, which all induce 3'-phosphoglycolate-terminated double strand breaks. PMID- 17121862 TI - ATP6 homoplasmic mutations inhibit and destabilize the human F1F0-ATP synthase without preventing enzyme assembly and oligomerization. AB - The molecular pathogenic mechanism of the human mitochondrial diseases neurogenic ataxia and retinitis pigmentosa and maternally inherited Leigh syndrome was determined in cultured human cells harboring homoplasmic T8993G/T8993C point mutations in the mitochondrial ATP6 gene, which encodes subunit 6 of the F1F0-ATP synthase. Immunoprecipitation and blue native electrophoresis showed that F1F0 ATP synthase assembles correctly in homoplasmic mutant mitochondria. The mutants exhibited a tendency to have an increased sensitivity to subsaturating amounts of oligomycin; this provided further evidence for complete assembly and tight coupling between the F1 and F0 sectors. Furthermore, human ATP synthase dimers and higher homo-oligomers were observed for the first time, and it was demonstrated that the mutant enzymes retain enough structural integrity to oligomerize. A reproducible increase in the proportion of oligomeric-to-monomeric enzyme was found for the T8993G mutant suggesting that F1F0 oligomerization is regulated in vivo and that it can be modified in pathological conditions. Despite correct assembly, the T8993G mutation produced a 60% inhibition in ATP synthesis turnover. In vitro denaturing conditions showed F1F0 instability conferred by the mutations, although this instability did not produce enzyme disassembly in the conditions used for determination of ATP synthesis. Taken together, the data show that the primary molecular pathogenic mechanism of these deleterious human mitochondrial mutations is functional inhibition in a correctly assembled ATP synthase. Structural instability may play a role in the progression of the disease under potentially denaturing conditions, as discussed. PMID- 17121863 TI - Regulation of Polo-like kinase 1 by DNA damage in mitosis. Inhibition of mitotic PLK-1 by protein phosphatase 2A. AB - DNA damage triggers multiple checkpoint pathways to arrest cell cycle progression. Polo-like kinase 1 (Plk1) is an important regulator of several events during mitosis. In addition to Plk1 functions in cell cycle, Plk1 is involved in DNA damage check-point in G2 phase. Normally, ataxia telangiectasia mutated kinase (ATM) is a key enzyme involved in G2 phase cell cycle arrest following DNA damage, and inhibition of Plk1 by DNA damage during G2 occurs in a ATM/ATR-dependent manner. However, it is still unclear how Plk1 is regulated in response to DNA damage in mitosis in which Plk1 is already activated. Here, we show that treatment of mitotic cells with doxorubicin and gamma-irradiation inhibits Plk1 activity through dephosphorylation of Plk1, and cells were arrested in G2 phase. Treatments of the phosphatase inhibitors and siRNA experiments suggested that PP2A pathway might be involved in regulating mitotic Plk1 activity in mitotic DNA damage. Finally, we propose a novel pathway, which is connected between ATM/ATR/Chk and protein phosphatase-Plk1 in DNA damage response in mitosis. PMID- 17121864 TI - Marginal modeling of nonnested multilevel data using standard software. AB - Epidemiologic data are often clustered within multiple levels that may not be nested within each other. Generalized estimating equations are commonly used to adjust for correlation among observations within clusters when fitting regression models; however, standard software does not currently accommodate nonnested clusters. This paper introduces a simple generalized estimating equation strategy that uses available commercial or public software for the regression analysis of nonnested multilevel data. The authors describe how to obtain empirical standard error estimates for constructing valid confidence intervals and conducting statistical hypothesis tests. The method is evaluated using simulations and illustrated with an analysis of data from the Breast Cancer Surveillance Consortium that estimates the influence of woman, radiologist, and facility characteristics on the positive predictive value of screening mammography. Performance with a small number of clusters is discussed. Both the simulations and the example demonstrate the importance of accounting for the correlation within all levels of clustering for proper inference. PMID- 17121865 TI - RhoA activation in mesangial cells by mechanical strain depends on caveolae and caveolin-1 interaction. AB - Increased intraglomerular pressure is an important hemodynamic determinant of glomerulosclerosis and can be modeled in vitro by exposing mesangial cells to cyclic mechanical strain. A previous study showed that RhoA mediates strain induced production of fibronectin; herein is investigated the role of caveolae in RhoA activation. Cyclodextrin and filipin, agents that disrupt caveolae, abrogated strain-induced RhoA activation in mesangial cells. Caveolin-1 (cav-1), the defining protein of caveolae, was Y14 phosphorylated by strain, and this was inhibited by PP1, showing Src dependence. Strain also induced c-SrcY416 phosphorylation and hence activation. Strain increased RhoA association with cav 1, which was blocked by PP1. Cyclodextrin and filipin inhibited the strain induced RhoA/cav-1 association, indicating dependence on caveolar structural integrity. Restoration of caveolae by coincubation of cyclodextrin with cholesterol rescued both RhoA activation and RhoA/cav-1 association in response to strain. Sucrose gradient detected a significant portion of RhoA in caveolae, with Src located exclusively in these domains. Finally, in cells that were infected with retrovirus that encodes the nonphosphorylatable cav-1 Y14A, RhoA/cav-1 association, RhoA activation, and fibronectin secretion in response to strain were abrogated. It is concluded that strain-induced RhoA activation depends on the integrity of caveolae and on physical association of cav-1 and RhoA. The phosphorylation of cav-1 at Y14 by Src kinases is required for this to occur. These studies define a novel function for cav-1 and caveolae as positive effectors of RhoA activation. Targeting caveolae thus may provide a new therapeutic option for glomerular sclerosis that is associated with elevated intraglomerular pressure. PMID- 17121866 TI - Coregulator exchange and sphingosine-sensitive cooperativity of steroidogenic factor-1, general control nonderepressed 5, p54, and p160 coactivators regulate cyclic adenosine 3',5'-monophosphate-dependent cytochrome P450c17 transcription rate. AB - Transcription of the cytochrome P450 17 (CYP17) gene is regulated by cAMP dependent binding of steroidogenic factor-1 (SF-1) to its promoter in the adrenal cortex. Using temporal chromatin immunoprecipitation and mammalian two-hybrid experiments, we establish the reciprocal presence of coactivators [general control nonderepressed (GCN5), cAMP response element-binding protein-binding protein, p300, p300/cAMP response element-binding protein-binding protein CBP associated factor, p160s, polypyrimidine tract associated splicing factor, and p54(nrb)], corepressors (class I histone deacetylases, receptor interacting protein, nuclear receptor corepressor, and Sin3A), and SWI/SNF (human homolog of yeast mating type switching/sucrose nonfermenting) and imitation SWI chromatin remodeling ATPases on the CYP17 promoter during transcription cycles in the H295R adrenocortical cell line. A ternary GCN5/SRC-1/SF-1 complex forms on the CYP17 promoter with cAMP-dependence within 30 min of cAMP stimulation, and corresponds with SWI/SNF chromatin remodeling. This complex is sensitive to the SF-1 antagonist sphingosine and results in decreased transcription of CYP17. GCN5 acetyltransferase activity and carboxy terminus binding proteins alternatively mediate disassembly of the complex. This work establishes the temporal order of cAMP-induced events on the promoter of a key steroidogenic gene during SF-1 mediated transcription. PMID- 17121867 TI - Prospective comparison of three predictive rules for assessing severity of community-acquired pneumonia in Hong Kong. AB - BACKGROUND: Community-acquired pneumonia (CAP) is a leading infectious cause of death throughout the world, including Hong Kong. AIM: To compare the ability of three validated prediction rules for CAP to predict mortality in Hong Kong: the 20 variable Pneumonia Severity Index (PSI), the 6-point CURB65 scale adopted by the British Thoracic Society and the simpler CRB65. METHODS: A prospective observational study of 1016 consecutive inpatients with CAP (583 men, mean (SD) age 72 (17) years) was performed in a university hospital in the New Territories of Hong Kong in 2004. The patients were classified into three risk groups (low, intermediate and high) according to each rule. The ability of the three rules to predict 30 day mortality was compared. RESULTS: The overall mortality and intensive care unit (ICU) admission rates were 8.6% and 4.0%, respectively. PSI, CURB65 and CRB65 performed similarly, and the areas under the receiver operating characteristic (ROC) curve were 0.736 (95% CI 0.687 to 0.736), 0.733 (95% CI 0.679 to 0.787) and 0.694 (95% CI 0.634 to 0.753), respectively. All three rules had high negative predictive values but relatively low positive predictive values at all cut-off points. Larger proportions of patients were identified as low risk by PSI (47.2%) and CURB65 (43.3%) than by CRB65 (12.6%). CONCLUSION: All three predictive rules have a similar performance in predicting the severity of CAP, but CURB65 is more suitable than the other two for use in the emergency department because of its simplicity of application and ability to identify low risk patients. PMID- 17121868 TI - Randomised study of three non-surgical treatments in mild to moderate obstructive sleep apnoea. AB - BACKGROUND: Patients with mild to moderate obstructive sleep apnoea (OSA) may be managed with different treatment options. This study compared the effectiveness of three commonly used non-surgical treatment modalities. METHODS: Subjects with mild to moderate OSA were randomised to one of three treatment groups for 10 weeks: conservative measures (sleep hygiene) only, continuous positive airways pressure (CPAP) in addition to conservative measures or an oral appliance in addition to conservative measures. All overweight subjects were referred to a weight-reduction class. OSA was assessed by polysomnography. Blood pressure was recorded in the morning and evening in the sleep laboratory. Daytime sleepiness was assessed with the Epworth Sleepiness Scale. Health-related quality of life (HRQOL) was assessed with the 36-Item Short-Form Health Survey (SF-36) and Sleep Apnoea Quality of Life Index (SAQLI). RESULTS: 101 subjects with a mean (SEM) apnoea-hypopnoea index (AHI) of 21.4 (1.1) were randomised to one of the three groups. The severity of sleep-disordered breathing was decreased in the CPAP and oral appliance groups compared with the conservative measures group, and the CPAP group was significantly better than the oral appliance group. Relief from sleepiness was significantly better in the CPAP group. CPAP was also better than the oral appliance or conservative measures in improving the "bodily pain" domain, and better than conservative measures in improving the "physical function" domain of SF-36. Both CPAP and the oral appliance were more effective than conservative measures in improving the SAQLI, although no difference was detected between the CPAP and oral appliance groups. CPAP and the oral appliance significantly lowered the morning diastolic blood pressure compared with baseline values, but there was no difference in the changes in blood pressure between the groups. There was also a linear relationship between the changes in AHI and body weight. CONCLUSION: CPAP produced the best improvement in terms of physiological, symptomatic and HRQOL measures, while the oral appliance was slightly less effective. Weight loss, if achieved, resulted in an improvement in sleep parameters, but weight control alone was not uniformly effective. PMID- 17121869 TI - Physical activity and bronchial hyperresponsiveness: European Community Respiratory Health Survey II. AB - BACKGROUND: Identification of the risk factors for bronchial hyperresponsiveness (BHR) would increase the understanding of the causes of asthma. The relationship between physical activity and BHR in men and women aged 28.0-56.5 years randomly selected from 24 centres in 11 countries participating in the European Community Respiratory Health Survey II was investigated. METHODS: 5158 subjects answered questionnaires about physical activity and performed BHR tests. Participants were asked about the frequency and duration of usual weekly exercise resulting in breathlessness or sweating. BHR was defined as a decrease in forced expiratory volume in 1 s of at least 20% of its post-saline value for a maximum methacholine dose of 2 mg. RESULTS: Both frequency and duration of physical activity were inversely related to BHR. The prevalence of BHR in subjects exercising or=4 times a week was 14.5%, 11.6% and 10.9%, respectively (p<0.001). The corresponding odds ratios were 1.00, 0.78 (95% CI 0.62 to 0.99) and 0.69 (95% CI 0.50 to 0.94) after controlling for potential confounding factors. The frequency of BHR in subjects exercising <1 h, 1-3 h and >or=4 h a week was 15.9%, 10.9% and 10.7%, respectively (p<0.001). The corresponding adjusted odds ratios were 1.00, 0.70 (95% CI 0.57 to 0.87) and 0.67 (95% CI 0.50 to 0.90). Physical activity was associated with BHR in all studied subgroups. CONCLUSIONS: These results suggest that BHR is strongly and independently associated with decreased physical activity. Further studies are needed to determine the mechanisms underlying this association. PMID- 17121870 TI - Early detection of cystic fibrosis lung disease: multiple-breath washout versus raised volume tests. AB - BACKGROUND: Lung clearance index (LCI), a measure of ventilation inhomogeneity derived from the multiple-breath inert gas washout (MBW) technique, has been shown to detect abnormal lung function more readily than spirometry in preschool children with cystic fibrosis, but whether this holds true during infancy is unknown. OBJECTIVES: To compare the extent to which parameters derived from the MBW and the raised lung volume rapid thoraco-abdominal compression (RVRTC) techniques identify diminished airway function in infants with cystic fibrosis when compared with healthy controls. METHODS: Measurements were performed during quiet sleep, with the tidal breathing MBW technique being performed before the forced expiratory manoeuvres. RESULTS: Measurements were obtained in 39 infants with cystic fibrosis (mean (SD) age 41.4 (22.0) weeks) and 21 controls (37.0 (15.1) weeks). Infants with cystic fibrosis had a significantly higher respiratory rate (38 (10) vs 32 (5) bpm) and LCI (8.4 (1.5) vs 7.2 (0.3)), and significantly lower values for all forced expiratory flow-volume parameters compared with controls. Girls with cystic fibrosis had significantly lower forced expiratory volume (FEV(0.5) and FEF(25-75 )) than boys (mean (95% CI girls-boys): -1.2 (-2.1 to -0.3) for FEV(0.5) Z score; FEF(25-75): -1.2 (-2.2 to -0.15)). When using both the MBW and RVRTC techniques, abnormalities were detected in 72% of the infants with cystic fibrosis, with abnormalities detected in 41% using both techniques and a further 15% by each of the two tests performed. CONCLUSIONS: These findings support the view that inflammatory and/or structural changes in the airways of children with cystic fibrosis start early in life, and have important implications regarding early detection and interventions. Monitoring of early lung disease and functional status in infants and young children with cystic fibrosis may be enhanced by using both MBW and the RVRTC. PMID- 17121871 TI - Multilocus analysis of atopy in Korean children using multifactor-dimensionality reduction. AB - BACKGROUND: Atopy is considered to be a complex genetic trait and does not follow a simple mendelian pattern of inheritance. It is now well recognised that gene gene interactions are important in complex genetic disease. AIM: To analyse the influence of gene-gene interactions in the development of atopy. METHODS: A total of 2055 ethnically identical participants aged 10-18 years living in rural areas on Jeju Island, Korea, were randomly recruited. Atopy was defined as a positive skin prick test response to one or more common inhalant allergens. Gene-gene interactions among 12 polymorphic loci were analysed in the seven candidate genes of atopy using the multidimensionality-reduction method. RESULTS: A significant interaction was found between V297I in the gene coding vascular endothelial growth factor receptor 2 (KDR) and -308G-->A in the gene coding tumour necrosis factor (TNF)alpha on the risk of atopy, with a cross-validation consistency of 10 out of 10 and a prediction error of 35.9% (p = 0.001). Conventional logistic regression also revealed significant interactions between KDR and TNF for atopy. Individuals with the variant allele of -308G-->A in TNF (GA or AA) and V297I in KDR (VI or II) had a significantly higher risk of atopy (OR 2.23; 95% CI 1.48 to 3.57). CONCLUSION: KDR and TNF may synergistically influence the development of atopy through gene-gene interaction in Korean children and adolescents. PMID- 17121872 TI - Use of inhaled corticosteroids during the first trimester of pregnancy and the risk of congenital malformations among women with asthma. AB - AIM: To investigate whether the maternal use of different doses of inhaled corticosteroids (ICSs) during the first trimester of pregnancy for the treatment of asthma increases the risk of congenital malformations in the offspring. METHODS: From the linkage of three administrative Canadian databases, a cohort of 4561 pregnancies from women with asthma who delivered between 1990 and 2000 was reconstructed. A two-stage sampling cohort design was used to acquire additional data from the woman's medical chart. Cases of congenital malformation were identified from the medical services database or the hospital database. Using refill patterns of medications, the average daily dose of ICSs used during the first trimester was calculated and categorised as follows: 0, 1-500, 500-1000 and >1000 microg/day in beclomethasone-chlorofluorocarbon equivalent. A Generalized Estimation Equation model was used to estimate the adjusted odds ratio of congenital malformation as a function of ICS daily dose. All analyses were performed for all malformations and major malformations separately. RESULTS: Within the cohort 418 babies were identified with a congenital malformation (9.2%), 278 of which had a major malformation. About 40% of women used ICSs during the first trimester, but only 5.3% of women used >500 microg/day. The adjusted odds ratio (95% CI) for all malformations associated with the use of ICSs during the first trimester was: 0.77 (0.53 to 1.13) for 1-500, 0.41 (0.19 to 0.92) for 501-1000 and 1.00 (0.42 to 2.36) for >1000 microg/day. The corresponding figures for major malformations were 0.90 (0.64 to 1.24), 0.56 (0.22 to 1.43) and 1.67 (0.56 to 5.03). CONCLUSION: This study adds evidence to the safety of ICSs for the treatment of asthma during pregnancy, with regard to the likelihood of congenital malformation. PMID- 17121873 TI - A neoadjuvant/adjuvant randomized trial of colorectal cancer patients vaccinated with an anti-idiotypic antibody, 105AD7, mimicking CD55. AB - PURPOSE: To assess the tolerability and effectiveness of 105AD7 vaccination in colorectal cancer patients. 105AD7 is a human anti-idiotypic antibody mimicking CD55, a glycoprotein, which is more than expressed on colorectal cancer cells and protects them from attack by complement. EXPERIMENTAL DESIGN: Colorectal cancer patients (n = 67) eligible for primary surgery were randomized to receive the anti-idiotypic antibody 105AD7+/-Bacillus Calmette-Guerin/alum or to no treatment (control group). The immunizations were given i.d./i.m. before surgery and continued for a period of 2 years. The patients were monitored in enzyme-linked immunospot (ELISPOT; gamma-IFN), proliferation assay, and Luminex cytokine assays. RESULTS: No serious adverse events were recorded. Of the 32 investigated immunized patients, 14 (44%) were considered to be responders in the ELISPOT assay. Induced proliferative responses were noted in 17 of 40 (43%) monitored patients. There was no correlation between the ELISPOT and proliferation assays. Luminex analyses revealed tumor necrosis factor-alpha and granulocyte macrophage colony-stimulating factor responses not only to the vaccine but also toward the native antigen CD55 in 9 of 13 (69%) patients. CONCLUSIONS: Immune responses to vaccination were induced in a majority of monitored patients measured by ELISPOT and proliferation assay. The lack of correlation between the ELISPOT and proliferation assays may reflect the fact that the two methods measure different T-cell responses and highlights the importance of multiple readouts in evaluating a potential cancer vaccine. Responses to both the anti-idiotype and the CD55 antigen were measurable, adding support to the use of CD55 as a target in cancer treatment. PMID- 17121874 TI - RNASEN regulates cell proliferation and affects survival in esophageal cancer patients. AB - PURPOSE: MicroRNAs (miRNA) are small noncoding RNAs thought to be involved in physiologic and developmental processes by negatively regulating the expression of target genes. Little is known about the role of miRNAs in normal and cancer cells. It is possible that deregulation of miRNA may contribute to the oncogenesis of some cancers. We studied the expression level of the miRNA processing enzyme (DICER1, DGCR8, and RNASEN) in esophageal squamous cell carcinoma (ESCC). EXPERIMENTAL DESIGN: The expression levels of DICER1, DGCR8, and RNASEN mRNA in 73 ESCC tissues were compared with that in corresponding normal esophageal epithelium by Taqman real-time reverse-transcription PCR. We also examined RNASEN protein expression in 27 cell lines. The role of RNASEN in cell proliferation in ESCC cells was assessed by small interfering RNA. Paraffin sections of ESCC patients were immunohistochemically investigated. RESULTS: We found that RNASEN expression levels were enhanced in a fraction of esophageal cancers. Multivariate Cox regression analysis showed that the prognostic effect of RNASEN (P = 0.0036) seems to be independent of disease stage (P = 0.0060). Knockdown of RNASEN in esophageal cancer cell lines resulted in a 46% to 85% reduction in cell number. In an immunohistochemical study, the intensity of RNASEN expression was often increased in the tumor compared with that in normal epithelium. CONCLUSIONS: The relationship between the RNASEN expression and the prognosis of the ESCC patients warrants a further study on the role of miRNA and tumor progression. PMID- 17121875 TI - Epigenetic biomarkers and breast cancer: cause for optimism. PMID- 17121876 TI - Fluorescence visualization in oral neoplasia: shedding light on an old problem. PMID- 17121877 TI - Of spiders and crabs: the emergence of lysophospholipids and their metabolic pathways as targets for therapy in cancer. AB - Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P), two small lysophospholipids, are potent inducers of many of the hallmarks of cancer including cell proliferation, survival, migration, invasion, and neovascularization in in vitro and in vivo tumor models. Furthermore, the enzymes metabolizing LPA and S1P and their receptors are aberrant in multiple cancer lineages and exhibit transforming activity altering patterns and targets for metastasis. Several recent studies show the remarkable activity of new chemical genomics and/or potential novel drugs in preclinical models. Combined with the physiologic and pathophysiologic activities of LPA and S1P, these studies suggest the implementation of preclinical and clinical evaluation of LPA and S1P as therapeutic targets. PMID- 17121878 TI - Synopsis of a roundtable on validating novel therapeutics for multiple myeloma. AB - PURPOSE: With the identification of new molecular targets and pathways, many new therapeutic approaches are being identified for potential application in the treatment of multiple myeloma. New chemical compounds and biologics have been developed against molecular targets with substantial scientific evidence that these targets are involved in myeloma development, progression, or relapse. To safely and rapidly bring these advances to bear on the disease, new preclinical models in cells and animals need to be established, as well as prioritization and standardization in current preclinical and clinical validation. An experts' roundtable was convened in November 2005 to discuss shortcomings in current preclinical models and discuss what models are needed to best validate therapeutics and combinations of therapies for multiple myeloma. CONCLUSIONS: This exciting event brought together experts in compound validation, preclinical development, and experts in multiple myeloma from academic institutions and the pharmaceutical and biotechnology industries. The goals were to evaluate an algorithm for therapeutic validation and discuss in vitro modeling for target discovery, animal models for preclinical development, and models for testing drug combinations. PMID- 17121879 TI - Enhanced RASGEF1A expression is involved in the growth and migration of intrahepatic cholangiocarcinoma. AB - PURPOSE AND EXPERIMENTAL DESIGN: To identify novel molecular targets for the treatment of intrahepatic cholangiocarcinoma (ICC), the second most common type of primary hepatobiliary cancer, we earlier analyzed genome-wide expression profiles of genes in 25 ICCs. Among the genes whose expression levels were commonly elevated in the tumors, we identified a novel gene termed RASGEF1A that encodes a putative Ras guanine nucleotide exchange factor domain-containing protein. RESULTS: We showed in this article that RASGEF1A protein has a guanine nucleotide exchange activity to K-RAS, H-RAS, and N-RAS proteins in vitro. Consistently, exogenous RASGEF1A expression increased the activity of Ras. In addition, suppression of RASGEF1A by small interfering RNA retarded the growth of cholangiocarcinoma cells. Interestingly, COS7 cells expressing exogenous RASGEF1A showed enhanced cellular motility in Transwell and wound-healing assays. CONCLUSIONS: These data suggest that elevated expression of RASGEF1A may play an essential role for proliferation and progression of ICC. Our data indicate that RASGEF1A may be a promising therapeutic target for the majority of ICCs. PMID- 17121880 TI - Identification of carboxypeptidase of glutamate like-B as a candidate suppressor in cell growth and metastasis in human hepatocellular carcinoma. AB - PURPOSE: We have previously done large-scale cDNA transfection screening on human hepatocellular carcinoma (HCC) cells and have identified 3,806 cDNA genes that possess the ability of either stimulating or inhibiting cell growth. In this study, we characterized one of these growth suppressor genes, carboxypeptidase of glutamate like-B (CPGL-B), in HCC. EXPERIMENTAL DESIGN: Semiquantitative reverse transcription PCR was used to examine the expression levels of CPGL-B. The cellular localization and functions of CPGL-B were investigated by enforced expression of CPGL-B in HCC cells. RESULTS: From our previous cDNA transfection screening, we identified a gene named CPGL and its isoform, CPGL-B. With computational analysis, CPGL was located at chromosome 18q22.3 and was a homologue of peptidase family M20. CPGL was expressed in all adult and fetal tissues, whereas its isoform, CPGL-B, lacking exons 3 and 4, was expressed in all fetal tissues but only in liver and placenta of adult tissues. In HCC, CPGL-B was frequently underexpressed (35 of 90, 38.9%) in tumorous tissues compared with the corresponding nontumorous livers. Intriguingly, the underexpression was significantly associated with the presence of venous invasion (P=0.018) and tumor microsatellite formation (P=0.004). Stable transfection of CPGL-B in SMMC7721 HCC cells showed significant inhibition in cell viability, colony formation, cell invasion, and tumor formation in nude mice. CPGL-B also down-regulated CXCR3, matrix metalloproteinase 11, and CD44s, which are involved in cell growth and cell migration. CONCLUSIONS: These findings suggest that the frequent underexpression of CPGL-B may be associated with cell growth and metastasis of HCC. PMID- 17121881 TI - Mapping geographic zones of cancer risk with epigenetic biomarkers in normal breast tissue. AB - PURPOSE: Genetic alterations were previously identified in normal epithelia adjacent to invasive cancers. The aim of this study was to determine DNA methylation in histologically normal tissues from multiple geographic zones adjacent to primary breast tumors. EXPERIMENTAL DESIGN: First, methylation status of a 4-kb region of RASSF1A promoter was interrogated using oligonucleotide-based microarray in 144 samples (primary tumors, 47; adjacent normals, 69; reduction mammoplasty tissues, 28). Second, allelic imbalance (AI)/loss of heterozygosity (LOH) surrounding RASSF1A promoter were analyzed in 30 samples (tumors, 8; adjacent normals, 22). Third, global methylation screening of 49 samples (tumors, 12; adjacent normals, 25; reduction mammoplasty, 12) was done by differential methylation hybridization. Real-time quantitative methylation-specific PCR was used to validate the microarray findings. RESULTS: DNA methylation in the core RASSF1A promoter was low in reduction mammoplasty tissues (P=0.0001) when compared with primary tumors. The adjacent normals had an intermediate level of methylation. The regions surrounding the core were highly methylated in all sample types. Microsatellite markers showed AI/LOH in tumors and some of the adjacent normals. Concurrent AI/LOH and DNA methylation in RASSF1A promoter occurred in two of six tumors. Global methylation screening uncovered genes more methylated in adjacent normals than in reduction mammoplasty tissues. The methylation status of four genes was confirmed by quantitative methylation specific PCR. CONCLUSIONS: Our findings suggest a field of methylation changes extending as far as 4 cm from primary tumors. These frequent alterations may explain why normal tissues are at risk for local recurrence and are useful in disease prognostication. PMID- 17121882 TI - Reprimo methylation is a potential biomarker of Barrett's-Associated esophageal neoplastic progression. AB - PURPOSE: Reprimo, a candidate tumor-suppressor gene, regulates p53-mediated cell cycle arrest at G2 phase, and tumor-suppressor gene methylation is involved in the pathogenesis and progression of esophageal cancer. Our aim was to determine whether and at what phase of neoplastic progression Reprimo methylation occurs in Barrett's adenocarcinogenesis, as well as its columnar or squamous cell-type specificity. We also sought to determine whether Reprimo expression could be restored in vitro by the demethylating agent 5-aza-deoxycytidine (5AzaC). EXPERIMENTAL DESIGN: Quantitative methylation-specific PCR for Reprimo was done using an ABI7700 (Taqman) apparatus on 175 endoscopic biopsy specimens. In addition, reverse transcription-PCR and quantitative methylation-specific PCR were done on esophageal carcinoma cells before and after treatment with 5AzaC. RESULTS: In Barrett's esophagus (BE; P=0.001), high-grade dysplasia (HGD; P=0.001), and esophageal adenocarcinoma (EAC; P=0.00003), the level and frequency of Reprimo methylation were significantly higher than in normal esophagus (NE). There was no statistically significant difference between BE and EAC, HGD and EAC, or NE and esophageal squamous cell carcinoma (ESCC). Reprimo methylation occurred in 0 of 19 NE samples, 6 (13%) of 45 ESCC, 9 (36%) of 25 BE, 7 (64%) of 11 HGD, and 47 (63%) of 75 EAC. Analysis of Reprimo methylation in EAC versus NE revealed an area under the receiver-operator characteristic curve of 0.812 (P<0.00001; 95% confidence interval, 0.73-0.90). In vitro 5AzaC treatment of OE33 EAC cells reduced Reprimo methylation and increased Reprimo expression. CONCLUSIONS: Reprimo methylation occurs significantly more frequently in BE, HGD, and EAC than in NE or ESCC, suggesting that this epigenetic alteration is a specialized columnar, cell-specific early event with potential as a biomarker for the early detection of esophageal neoplasia. PMID- 17121883 TI - Molecular and cytogenetic subgroups of oropharyngeal squamous cell carcinoma. AB - PURPOSE: The aim of this study was to acquire further insights into the pathogenetic pathways of head and neck squamous cell carcinomas (HNSCC) that may be useful for identifying new biomarkers instrumental in developing more specific treatment approaches. EXPERIMENTAL DESIGN: Cell cycle regulators and epidermal growth factor receptor (EGFR) and BRAF genes were analyzed in a series of 90 oropharyngeal SCCs of a cohort of surgically treated patients from a single institution, and the results were matched with the presence of high-risk human papillomavirus (HR-HPV) DNA and the TP53 status. RESULTS: At least four distinct groups of tumors were identified sharing a common histology but displaying different molecular/cytogenetic patterns: (a) 19% were HPV-positive SCCs whose lack of alterations of the investigated genes could explain their particular natural history, which requires less aggressive treatment; (b) 37% were HPV negative SCCs carrying TP53 mutations, which may be more effectively treated by drugs acting through p53-independent apoptosis; (c) 34% were HPV-negative SCCs carrying wild-type TP53 and loss of 9p21 (p16INK4a and p15INK4b) and/or cyclin D1 overexpression that justify treatment with DNA-damaging drugs followed by cell cycle inhibitors; and (d) 10% were HPV-negative lacking tumor suppressor genes and cell cycle alterations. The second, third, and fourth groups also showed an increased copy number of EGFR and chromosome 7 (43%) that might justify the additional or alternative use of EGFR inhibitors. CONCLUSIONS: Our findings suggest that assessing HPV, TP53, 9p21, and EGFR status may be crucial to finding more tailored and beneficial treatments for oropharyngeal SCCs. PMID- 17121884 TI - FGFR1 emerges as a potential therapeutic target for lobular breast carcinomas. AB - PURPOSE: Classic lobular carcinomas (CLC) account for 10% to 15% of all breast cancers. At the genetic level, CLCs show recurrent physical loss of chromosome16q coupled with the lack of E-cadherin (CDH1 gene) expression. However, little is known about the putative therapeutic targets for these tumors. The aim of this study was to characterize CLCs at the molecular genetic level and identify putative therapeutic targets. EXPERIMENTAL DESIGN: We subjected 13 cases of CLC to a comprehensive molecular analysis including immunohistochemistry for E cadherin, estrogen and progesterone receptors, HER2/neu and p53; high-resolution comparative genomic hybridization (HR-CGH); microarray-based CGH (aCGH); and fluorescent and chromogenic in situ hybridization for CCND1 and FGFR1. RESULTS: All cases lacked the expression of E-cadherin, p53, and HER2, and all but one case was positive for estrogen receptors. HR-CGH revealed recurrent gains on 1q and losses on 16q (both, 85%). aCGH showed a good agreement with but higher resolution and sensitivity than HR-CGH. Recurrent, high level gains at 11q13 (CCND1) and 8p12-p11.2 were identified in seven and six cases, respectively, and were validated with in situ hybridization. Examination of aCGH and the gene expression profile data of the cell lines, MDA-MB-134 and ZR-75-1, which harbor distinct gains of 8p12-p11.2, identified FGFR1 as a putative amplicon driver of 8p12-p11.2 amplification in MDA-MB-134. Inhibition of FGFR1 expression using small interfering RNA or a small-molecule chemical inhibitor showed that FGFR1 signaling contributes to the survival of MDA-MB-134 cells. CONCLUSIONS: Our findings suggest that receptor FGFR1 inhibitors may be useful as therapeutics in a subset of CLCs. PMID- 17121885 TI - Nomograms provide improved accuracy for predicting survival after radical cystectomy. AB - AIMS: To develop multivariate nomograms that determine the probabilities of all cause and bladder cancer-specific survival after radical cystectomy and to compare their predictive accuracy to that of American Joint Committee on Cancer (AJCC) staging. METHODS: We used Cox proportional hazards regression analyses to model variables of 731 consecutive patients treated with radical cystectomy and bilateral pelvic lymphadenectomy for bladder transitional cell carcinoma. Variables included age of patient, gender, pathologic stage (pT), pathologic grade, carcinoma in situ, lymphovascular invasion (LVI), lymph node status (pN), neoadjuvant chemotherapy (NACH), adjuvant chemotherapy (ACH), and adjuvant external beam radiotherapy (AXRT). Two hundred bootstrap resamples were used to reduce overfit bias and for internal validation. RESULTS: During a mean follow-up of 36.4 months, 290 of 731 (39.7%) patients died; 196 of 290 patients (67.6%) died of bladder cancer. Actuarial all-cause survival estimates were 56.3% [95% confidence interval (95% CI), 51.8-60.6%] and 42.9% (95% CI, 37.3-48.4%) at 5 and 8 years after cystectomy, respectively. Actuarial cancer-specific survival estimates were 67.3% (62.9-71.3%) and 58.7% (52.7-64.2%) at 5 and 8 years, respectively. The accuracy of a nomogram for prediction of all-cause survival (0.732) that included patient age, pT, pN, LVI, NACH, ACH, and AXRT was significantly superior (P=0.001) to that of AJCC staging-based risk grouping (0.615). Similarly, the accuracy of a nomogram for prediction of cancer-specific survival that included pT, pN, LVI, NACH, and AXRT (0.791) was significantly superior (P=0.001) to that of AJCC staging-based risk grouping (0.663). CONCLUSIONS: Multivariate nomograms provide a more accurate and relevant individualized prediction of survival after cystectomy compared with conventional prediction models, thereby allowing for improved patient counseling and treatment selection. PMID- 17121886 TI - Vascular targeted nanoparticles for imaging and treatment of brain tumors. AB - PURPOSE: Development of new therapeutic drug delivery systems is an area of significant research interest. The ability to directly target a therapeutic agent to a tumor site would minimize systemic drug exposure, thus providing the potential for increasing the therapeutic index. EXPERIMENTAL DESIGN: Photodynamic therapy (PDT) involves the uptake of a sensitizer by the cancer cells followed by photoirradiation to activate the sensitizer. PDT using Photofrin has certain disadvantages that include prolonged cutaneous photosensitization. Delivery of nanoparticles encapsulated with photodynamic agent specifically to a tumor site could potentially overcome the drawbacks of systemic therapy. In this study, we have developed a multifunctional polymeric nanoparticle consisting of a surface localized tumor vasculature targeting F3 peptide and encapsulated PDT and imaging agents. RESULTS: The nanoparticles specifically bound to the surface of MDA-435 cells in vitro and were internalized conferring photosensitivity to the cells. Significant magnetic resonance imaging contrast enhancement was achieved in i.c. rat 9L gliomas following i.v. nanoparticle administration. Serial magnetic resonance imaging was used for determination of pharmacokinetics and distribution of nanoparticles within the tumor. Treatment of glioma-bearing rats with targeted nanoparticles followed by PDT showed a significant improvement in survival rate when compared with animals who received PDT after administration of nontargeted nanoparticles or systemic Photofrin. CONCLUSIONS: This study reveals the versatility and efficacy of the multifunctional nanoparticle for the targeted detection and treatment of cancer. PMID- 17121887 TI - Frequent epigenetic inactivation of spleen tyrosine kinase gene in human hepatocellular carcinoma. AB - PURPOSE: The aim of present study was to investigate the methylation and expression status of spleen tyrosine kinase (SYK) in human hepatocellular carcinoma (HCC) and to evaluate this information for its ability to predict disease prognosis. E-cadherin and TIMP-3 methylation was also analyzed here as control because both were associated with poor prognosis in some types of tumors. EXPERIMENTAL DESIGN: We analyzed the methylation status of SYK, E-cadherin, and TIMP-3 in 124 cases of HCC and assessed the correlation of such methylations with clinicopathologic variables and prognosis after tumor resection. RESULTS: We found that SYK, E-cadherin, and TIMP-3 genes were methylated in 27%, 27%, and 42% of HCC neoplastic tissues, respectively. The loss of SYK mRNA or Syk protein expression was highly correlated with SYK gene methylation. The patients with methylated SYK in neoplastic tissues had a significantly lower overall survival rate after hepatectomy than those with unmethylated SYK. No significant difference in overall survival rates, however, was found between groups of patients with methylated and unmethylated E-cadherin or TIMP-3. Patients with negative Syk protein expression had a significantly lower overall survival rate than those with positive Syk protein expression. Multivariate analyses indicated that factors affecting overall survival were tumor-node-metastasis stage, Child Pugh classification, SYK methylation, or Syk protein status. CONCLUSIONS: Our results indicate that SYK methylation and loss of Syk expression in HCC neoplastic tissues are independent biomarkers of poor patient outcome and that determination of SYK methylation or Syk expression status may offer guidance for selecting appropriate treatments. PMID- 17121888 TI - Axillary lymph node nanometastases are prognostic factors for disease-free survival and metastatic relapse in breast cancer patients. AB - PURPOSE: Early breast cancer presents with a remarkable heterogeneity of outcomes. Undetected, microscopic lymph node tumor deposits may account for a significant fraction of this prognostic diversity. Thus, we systematically evaluated the presence of lymph node tumor cell deposits 5,000 nmol/L) against a panel of >120 unrelated kinases. In vitro, R547 effectively inhibited the proliferation of tumor cell lines independent of multidrug resistant status, histologic type, retinoblastoma protein, or p53 status, with IC(50)s PCI-2052 > PCI-2051 > PCI-2053. Thus, subtle changes in the alkyl substituents on the bipyrrole moiety result in significant changes in antitumor activity. PCI-2050 displayed significant antitumor efficacy in both Ramos and RKO xenograft models without hematologic, hepatic, or renal abnormalities as assessed by complete blood counts and serum chemistries. On the basis of these findings, it is concluded that the sapphyrin PCI-2050 warrants further evaluation as a potential anticancer agent due to its intrinsic proapoptotic activity and tumor localization ability. PMID- 17121927 TI - QW-1624F2-2, a synthetic analogue of 1,25-dihydroxyvitamin D3, enhances the response to other deltanoids and suppresses the invasiveness of human metastatic breast tumor cells. AB - The enzyme 24-hydroxylase, also known as CYP24, metabolizes 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] and is an established marker of vitamin D activity. Our studies evaluated the influence of a low-calcemic 1,25(OH)(2)D(3) analogue, QW 1624F2-2 (QW), on the regulation of CYP24 expression in MKL-4 cells, a metastatic mammary tumor cell model. 1,25(OH)(2)D(3) and its analogue, EB 1089, stimulated CYP24 induction at both protein and transcript levels. In contrast, QW failed to produce a sustained stimulation of CYP24, due, in large part, to a reduction in the stability of the CYP24 message. QW enhanced the capacity of 1,25(OH)(2)D(3) and EB 1089 to inhibit tumor cell proliferation by approximately 2-fold. QW also blocked the sustained induction of CYP24 expression by 1,25(OH)(2)D(3) and EB 1089, increased the potency of 1,25(OH)(2)D(3) and EB 1089, and inhibited breast tumor cell proliferation and invasion. PMID- 17121928 TI - Activation of mammalian target of rapamycin and the p70 S6 kinase by arsenic trioxide in BCR-ABL-expressing cells. AB - Arsenic trioxide (As(2)O(3)) exhibits important antitumor activities in vitro and in vivo, but the precise mechanisms by which it induces its effects are not known. We provide evidence that during treatment of BCR-ABL-expressing cells with As(2)O(3), there is activation of a cellular pathway involving the p70 S6 kinase (p70S6K). Our data show that p70S6K is rapidly phosphorylated on Thr(421) and Ser(424) and is activated in an As(2)O(3)-inducible manner. The mammalian target of rapamycin (mTOR) is also phosphorylated/activated in an As(2)O(3)-inducible manner, and its activity is required for downstream engagement of p70S6K. p70S6K subsequently phosphorylates the S6 ribosomal protein on Ser(235)/Ser(236) and Ser(240)/Ser(244) to promote initiation of mRNA translation. Treatment of chronic myelogenous leukemia-derived cell lines with As(2)O(3) also results in phosphorylation of the 4E-BP1 repressor of mRNA translation on Thr(37)/Thr(46) and Thr(70), sites required for its deactivation and its dissociation from the eukaryotic initiation factor 4E complex to allow cap-dependent mRNA translation. In studies to determine the functional relevance of this pathway, we found that inhibition of mTOR and downstream cascades enhances induction of apoptosis by As(2)O(3). Consistent with this, the mTOR inhibitor rapamycin strongly potentiated As(2)O(3)-mediated suppression of primitive leukemic progenitors from the bone marrow of chronic myelogenous leukemia patients. Altogether, our data show that the mTOR/p70S6K pathway is activated in a negative feedback regulatory manner in response to As(2)O(3) in BCR-ABL-transformed cells and plays a key regulatory role in the induction of anti-leukemic responses. PMID- 17121929 TI - Bcl-2 down-regulation and tubulin subtype composition are involved in resistance of ovarian cancer cells to vinflunine. AB - Vinflunine, a new microtubule-targeting drug, has a marked antitumor activity in vitro and in vivo. Here, we studied the mechanisms mediating resistance to vinflunine. We investigated the response to vinflunine of ovarian cancer cells initially selected as paclitaxel-resistant cells (A2780-TC1 cells). By comparison with A2780-wild-type (wt) cells, we showed that A2780-TC1 cells were highly resistant to vinflunine, with resistance factors reaching 800 and 1,830 for IC(50) and IC(70), respectively. We showed that P-glycoprotein minimally participated in this cell resistance. The examination of tubulin composition revealed increased levels of acetylated alpha-tubulin, betaII-tubulin, and betaIII-tubulin in A2780-TC1 cells before vinflunine treatment. As a consequence, vinflunine unequally affected microtubule network organization and function in A2780-wt and A2780-TC1 cells. Whereas the drug depolymerized microtubules and induced a mitotic block in A2780-wt cells, it did not depolymerize microtubules and induced a G(2) block in A2780-TC1 cells. Elsewhere, the mitochondrial protein Bcl-2 was down-regulated in A2780-TC1 cells. This down-regulation was related to resistance, as A2780-TC1 cells stably transfected with a Bcl-2 construct recovered a partial sensitivity to vinflunine. Lastly, we confirmed the role played by Bcl-2 by showing that the mitochondrial membrane potential was only disrupted by vinflunine in cells expressing Bcl-2. Altogether, our results indicate that modifications acquired during treatment (i.e., paclitaxel) have significant consequences on cell response to the following drug (i.e., vinflunine). Especially, this study shows that a specific pool of tubulin subtypes and a down-regulation of Bcl-2 are associated with resistance of ovarian cancer cells to vinflunine. PMID- 17121930 TI - Gallium-induced cell death in lymphoma: role of transferrin receptor cycling, involvement of Bax and the mitochondria, and effects of proteasome inhibition. AB - Gallium nitrate is a metallodrug with clinical efficacy in non-Hodgkin's lymphoma. Its mechanisms of antineoplastic action are not fully understood. In the present study, we investigated the roles of transferrin receptor (TfR) targeting and apoptotic pathways in gallium-induced cell death. Although DoHH2 lymphoma cells displayed a 3-fold lower number of TfRs than CCRF-CEM lymphoma cells, they were 3- to 4-fold more sensitive to gallium nitrate. Despite a lower TfR expression, DoHH2 cells had greater TfR cycling and iron and gallium uptake than CCRF-CEM cells. In other lymphoma cell lines, TfR levels per se did not correlate with gallium sensitivity. Cells incubated with gallium nitrate showed morphologic changes of apoptosis, which were decreased by the caspase inhibitor Z VAD-FMK and by a Bax-inhibitory peptide. Cells exposed to gallium nitrate released cytochrome c from mitochondria and displayed a dose-dependent increase in caspase-3 activity. An increase in active Bax levels without accompanying changes in Bcl-2 or Bcl-X(L) was seen in cells incubated with gallium nitrate. The endogenous expression of antiapoptotic Bcl-2 was greater in DoHH2 cells than in CCRF-CEM cells, suggesting that endogenous Bcl-2 levels do not correlate with cell sensitivity to gallium nitrate. Gallium-induced apoptosis was enhanced by the proteasome inhibitor bortezomib. Our results suggest that TfR function rather than TfR number is important in gallium targeting to cells and that apoptosis is triggered by gallium through the mitochondrial pathway by activating proapoptotic Bax. Our studies also suggest that the antineoplastic activity of combination gallium nitrate and bortezomib warrants further investigation. PMID- 17121931 TI - Pretherapy nuclear factor-kappaB status, chemoradiation resistance, and metastatic progression in esophageal carcinoma. AB - BACKGROUND: Transcriptional factor nuclear factor-kappaB (NF-kappaB) seems to be associated with aggressive clinical biology (chemoradiation resistance and metastatic progression) of esophageal cancer. We hypothesized that activated NF kappaB would define clinical biology irrespective of the type of chemotherapy or sequence administered. METHODS: Pretherapy and/or posttherapy cancer specimens were examined for activated NF-kappaB and correlated with pathologic response to chemoradiation, metastatic potential, overall survival, disease-free survival, and type of chemotherapy or sequence used. FINDINGS: Eighty patients undergoing chemotherapy and concurrent radiation were studied. Activated NF-kappaB prior to any therapy was associated with the lack of complete pathologic response (pathCR, P = 0.006). Forty-five (78%) of 58 patients achieving A, 2846A>T, 1679T>G, 85T>C, -1590T>C). Three hundred patients had no SNP. Forty-four patients had grade 3 to 4 toxic side effects in either the first or second cycle. Sixty percent of patients with either IVS14 + 1G>A or 2846A>T SNPs and the only patient with 1679T>G SNP experienced early grade 3 to 4 toxicity, compared with 0%, 5.5%, and 15% of those with either 1590T>C, 85T>C SNP, or no SNP, respectively. In cases with grade 3 to 4 toxicity, treatment either had to be quickly stopped, or could be safely continued with an individual dose adjustment. Sensitivity, specificity, and positive and negative predictive values of the detection of these three major SNPs as toxicity predictive factors were 0.31, 0.98, and 0.62 and 0.94, respectively. CONCLUSION: Pretreatment detection of three DPYD SNPs could help to avoid severe toxic side effects. This approach is suitable for clinical practice and should be compared or combined with pharmacologic approaches. In the case of dihydropyrimidine dehydrogenase deficiency, 5-FU administration often can be safely continued with an individual dose adjustment. PMID- 17121938 TI - Frequent overexpression of aurora B kinase, a novel drug target, in non-small cell lung carcinoma patients. AB - The serine/threonine protein kinase aurora B, a key regulator of mitosis, is emerging as a novel drug target for cancer treatment. Aurora B overexpression has been previously documented by immunohistochemistry in several types of human tumors. We assessed aurora B expression in a series of 160 non-small cell lung cancer (NSCLC) samples (60% stage I, 21% stage II, 11% stage III, and 8% stage IV). In addition, we determined the expression of survivin and p16, two molecules also involved in cell cycle control. Aurora B was expressed selectively in tumor cells compared with normal epithelium. Aurora B expression was significantly correlated with expression of survivin in the nucleus (P < 0.0001), but not with expression of p16 (P = 0.134). High aurora B expression levels were significantly associated with older age (P = 0.012), male sex (P = 0.013), squamous cell carcinoma histology (P = 0.001), poor tumor differentiation grade (P = 0.007), and lymph node invasion (P = 0.037), in the subset of radically resected patients in our series. In addition, aurora B expression predicted shorter survival for the patients with adenocarcinoma histology, at both univariate (P = 0.020) and multivariate (P = 0.012) analysis. Survivin expression levels were neither associated with patient clinicopathologic characteristics nor with survival. However, expression of survivin in the nucleus was preferentially detected in stage I and II than in stage III and IV (P = 0.007) in the overall series of NSCLC samples. Taken together, our results suggest that aurora B may represent a valid target in NSCLC. PMID- 17121939 TI - Gene expression profiles do not consistently predict the clinical treatment response in locally advanced breast cancer. AB - Neoadjuvant treatment offers an opportunity to correlate molecular variables to treatment response and to explore mechanisms of drug resistance in vivo. Here, we present a statistical analysis of large-scale gene expression patterns and their relationship to response following neoadjuvant chemotherapy in locally advanced breast cancers. We analyzed cDNA expression data from 81 tumors from two patient series, one treated with doxorubicin alone (51) and the other treated with 5 fluorouracil and mitomycin (30), and both were previously studied for correlations between TP53 status and response to therapy. We observed a low frequency of progressive disease within the luminal A subtype from both series (2 of 36 versus 13 of 45 patients; P = 0.0089) and a high frequency of progressive disease among patients with luminal B type tumors treated with doxorubicin (5 of 8 patients; P = 0.0078); however, aside from these two observations, no other consistent associations between response to chemotherapy and tumor subtype were observed. These specific associations could possibly be explained by covariance with TP53 mutation status, which also correlated with tumor subtype. Using supervised analysis, we could not uncover a gene profile that could reliably (>70% accuracy and specificity) predict response to either treatment regimen. PMID- 17121940 TI - C35 (C17orf37) is a novel tumor biomarker abundantly expressed in breast cancer. AB - Identification of shared tumor-specific targets is useful in developing broadly applicable therapies. In a study designed to identify genes up-regulated in breast cancer, a cDNA clone corresponding to a novel gene C35 (C17orf37) was selected by representational difference analysis of tumor and normal human mammary cell lines. Abundant expression of C35 transcript in tumors was confirmed by Northern blot and real-time PCR. The C35 gene is located on chromosome 17q12, 505 nucleotides from the 3' end of the ERBB2 oncogene, the antigenic target for trastuzumab (Herceptin) therapy. The chromosomal arrangement of the genes encoding C35 and ERBB2 is tail to tail. An open reading frame encodes a 12-kDa protein of unknown function. Immunohistochemical analysis detected robust and frequent expression of C35 protein, including 32% of grade 1 and 66% of grades 2 and 3 infiltrating ductal carcinomas of the breast (in contrast to 20% overexpressing HER-2/neu), 38% of infiltrating lobular carcinoma (typically HER 2/neu negative), as well as tumors arising in other tissues. C35 was not detected in 38 different normal human tissues, except Leydig cells in the testes and trace levels in a small percentage of normal breast tissue samples. The distinct and favorable expression profile of C35 spanning early through late stages of disease, including high frequency of overexpression in various breast carcinoma, abundant expression in distant metastases, and either absence or low level expression in normal human tissues, warrants further investigation of the relevance of C35 as a biomarker and/or a target for development of broadly applicable cancer-specific therapies. PMID- 17121941 TI - Benzyl isothiocyanate-induced apoptosis in human breast cancer cells is initiated by reactive oxygen species and regulated by Bax and Bak. AB - Epidemiologic studies have revealed an inverse correlation between dietary intake of cruciferous vegetables and the risk of breast cancer. We now show that cruciferous vegetable constituent benzyl isothiocyanate (BITC) effectively suppresses growth of cultured human breast cancer cells (MDA-MB-231 and MCF-7) by causing G(2)-M phase cell cycle arrest and apoptosis induction. On the other hand, a normal mammary epithelial cell line (MCF-10A) is significantly more resistant to growth arrest and apoptosis by BITC compared with breast cancer cells. The BITC-mediated cell cycle arrest was associated with a decrease in levels of proteins involved in regulation of G(2)-M transition, including cyclin B1, cyclin-dependent kinase 1, and cell division cycle 25C. The BITC-induced apoptosis correlated with induction of proapoptotic proteins Bax (MCF-7) and Bak (MDA-MB-231 and MCF-7) and down-regulation of antiapoptotic proteins Bcl-2 and Bcl-xL (MDA-MB-231). The SV40-immortalized mouse embryonic fibroblasts derived from Bax and Bak double knockout mice were significantly more resistant to BITC induced DNA fragmentation compared with wild-type mouse embryonic fibroblasts. The BITC treatment caused rapid disruption of the mitochondrial membrane potential, leading to cytosolic release of apoptogenic molecules, which was accompanied by formation of autophagosome-like structures as revealed by transmission electron microscopy. The BITC-mediated apoptosis was associated with generation of reactive oxygen species and cleavage of caspase-9, caspase-8, and caspase-3. Apoptosis induction by BITC was significantly attenuated in the presence of a combined superoxide dismutase and catalase mimetic EUK134 as well as caspase inhibitors. In conclusion, the present study reveals a complex signaling leading to growth arrest and apoptosis induction by BITC. PMID- 17121942 TI - Identification of SK3 channel as a new mediator of breast cancer cell migration. AB - Potassium channels have been involved in epithelial tumorigenesis but the role of small-conductance Ca(2+)-activated K(+) channels is unknown. We report here that small-conductance Ca(2+)-activated K(+) channels are expressed in a highly metastasizing mammary cancer cell line, MDA-MB-435s. Patch-clamp recordings showed typical small-conductance Ca(2+)-activated K(+) channel-mediated currents sensitive to apamin, 4-aminopyridine, and tetraethylammonium. Moreover, the cells displayed a high intracellular calcium concentration, which was decreased after 24 hours of apamin treatment. By regulating membrane potential and intracellular calcium concentration, these channels were involved in MDA-MB-435s cell migration, but not in proliferation. Only SK3 protein expression was observed in these cells in contrast to SK2, which was expressed both in cancer and noncancer cell lines. Whereas small interfering RNA directed against SK3 almost totally abolished MDA-MB-435s cell migration, transient expression of SK3 increased migration of the SK3-deficient cell lines, MCF-7 and 184A1. SK3 channel was solely expressed in tumor breast biopsies and not in nontumor breast tissues. Thus, SK3 protein channel seems to be a new mediator of breast cancer cell migration and represents a potential target for a new class of anticancer agents. PMID- 17121943 TI - Therapeutic potential of epidermal growth factor receptor-related protein. PMID- 17121945 TI - Noninvasive assessment of tumor vasculature response to radiation-mediated, vasculature-targeted therapy using quantified power Doppler sonography: implications for improvement of therapy schedules. AB - OBJECTIVE: Stereotactic radiotherapy (ablative radiation) is a modality that holds considerable promise for effective treatment of intracranial and extracranial malignancies. Although tumor vasculature is relatively resistant to small fractionated doses of ionizing radiation, large ablative doses of ionizing radiation lead to effective demise of the tumor vasculature. The purpose of this study was (1) to noninvasively monitor and compare tumor physiologic parameters in response to ablative radiation treatments and (2) to use these noninvasive parameters to optimize the schedule of administration of radiation therapy. METHODS: Lewis lung carcinoma tumors were implanted into C57BL/6 mice and treated with ablative radiation. The kinetics of change in physiologic parameters of a response to single-dose 20-Gy treatments was measured. Parameters studied included tumor blood flow, apoptosis, and proliferation rates. Serial tumor sections were stained to correlate noninvasive Doppler assessment of tumor blood flow with microvasculature histologic findings. RESULTS: A single administration of 20 Gy led to an incomplete tumor vascular response, with subsequent recovery of tumor blood flow within 4 days after treatment. Sustained reduction of tumor blood flow by administering the successive ablative radiation treatment before tumor blood flow recovery led to a 3-fold tumor growth delay. The difference in tumor volumes at each measurement time point (every 2 days) was statistically significant (P=.016). CONCLUSIONS: This study suggests a rational design of schedule optimization for radiation-mediated, vasculature-directed treatments guided by noninvasive assessment of tumor blood flow levels to ultimately improve the tumor response. PMID- 17121946 TI - Acoustic techniques for assessing the Optison destruction threshold. AB - OBJECTIVE: The purpose of this study was to identify the pressure threshold for the destruction of Optison (octafluoropropane contrast agent; Amersham Health, Princeton, NJ) using a laboratory-assembled 3.5-MHz pulsed ultrasound system and a clinical diagnostic ultrasound scanner. METHODS: A 3.5-MHz focused transducer and a linear array with a center frequency of 6.9 MHz were positioned confocally and at 90 degrees to each other in a tank of deionized water. Suspensions of Optison (5-8x10(4) microbubbles/mL) were insonated with 2-cycle pulses from the 3.5-MHz transducer (peak rarefactional pressure, or Pr, from 0.0, or inactive, to 0.6 MPa) while being interrogated with fundamental B-mode imaging pulses (mechanical index, or MI,=0.04). Scattering received by the 3.5-MHz transducer or the linear array was quantified as mean backscattered intensity or mean digital intensity, respectively, and fit with exponential decay functions (Ae-kt+N, where A+N was the amplitude at time 0; N, background echogenicity; and k, decay constant). By analyzing the decay constants statistically, a pressure threshold for Optison destruction due to acoustically driven diffusion was identified. RESULTS: The decay constants determined from quantified 3.5-MHz radio frequency data and B-mode images were in good agreement. The peak rarefactional pressure threshold for Optison destruction due to acoustically driven diffusion at 3.5 MHz was 0.15 MPa (MI=0.08). Furthermore, the rate of Optison destruction increased with increasing 3.5-MHz exposure pressure output. CONCLUSIONS: Optison destruction was quantified with a laboratory-assembled 3.5-MHz ultrasound system and a clinical diagnostic ultrasound scanner. The pressure threshold for acoustically driven diffusion was identified, and 3 distinct mechanisms of ultrasound contrast agent destruction were observed with acoustic techniques. PMID- 17121947 TI - Ultrasonographic differentiation of benign from malignant neck lymphadenopathy in thyroid cancer. AB - OBJECTIVE: The aim of this study was to determine whether ultrasonography itself was able to distinguish benign from malignant lymphadenopathy in patients with thyroid cancer. METHODS: We evaluated lymph nodes in a group of patients with thyroid cancer. Nodes were detected and measured by ultrasonography, and their shape, echogenicity, size, and location were noted. Ultrasonographically guided fine-needle aspiration biopsy (FNAB) was performed, and smears were analyzed cytologically. RESULTS: Ultrasonographically guided FNAB was performed in 578 neck nodes in a group of 631 patients with thyroid cancer. In most cases, metastases had a round shape and various echo structures, with a predomination of hypoechoic nodes without a hilum. There were statistical differences in size between metastatic and benign nodes in terms of maximum diameter, minimum diameter, and volume. Among these, minimum diameter and the shape of the nodes seemed to be the most reliable in suggesting malignancy. A round shape with a longitudinal/transverse ratio of less than 2 of hypoechoic nodes indicated the presence of metastases, and we then performed FNAB. The absence of an echogenic hilum and the presence of cystic portions and calcifications were significantly greater in malignancies than in benign lesions (P<.001). In most cases, metastatic nodules were situated in the lower third of the neck. Reactively enlarged nodes occurred more frequently in the upper part of the neck. CONCLUSIONS: Ultrasonography itself cannot distinguish benign from malignant lesions, but an echographic appearance suggests malignancy and helps in the selection of the node to aspirate with ultrasonographically guided FNAB, which is crucial for a final diagnosis. PMID- 17121948 TI - Cephalic vein measurement before forearm fistula creation: does use of a tourniquet to meet the venous diameter threshold increase the number of usable fistulas? AB - OBJECTIVE: To optimize forearm radiocephalic fistula success, many programs recommend a minimal cephalic vein diameter of 0.25 cm or greater on preoperative sonographic mapping. It is not established, however, whether a vein diameter before or after application of a tourniquet should be used in determining suitability for creation of a forearm fistula. METHODS: Before forearm radiocephalic fistula placement, preoperative sonographic mapping measured the cephalic vein diameter before and after application of a tourniquet. The patients fell into 2 groups: those with a pretourniquet vein diameter of 0.25 cm or greater (group 1) and those with a pretourniquet vein diameter of less than 0.25 cm that increased to 0.25 cm or greater after application of the tourniquet (group 2). The adequacy of each fistula for dialysis was determined clinically. RESULTS: Among 73 radiocephalic fistulas with known clinical outcomes, 28 were in group 1, and 45 were in group 2, on the basis of sonography. Fistula success rates were similar in group 1 patients (11 [39%] of 28) and group 2 patients (15 [33%] of 45) (P=.624, Fisher exact test). Inclusion of group 2 patients increased the number of patients recommended for placement of forearm fistulas and increased the total number of usable forearm fistulas from 11 to 26. The overall success rate of forearm fistulas was lower in women (19% versus 50%; P=.015, Fisher exact test). CONCLUSIONS: The use of a venous tourniquet increases the number of patients eligible for forearm fistulas without decreasing the adequacy rate of these fistulas. Therefore, a tourniquet should routinely be used in patients with small cephalic veins on pretourniquet evaluation. PMID- 17121949 TI - Bottle neck sign of the proximal portion of the internal carotid artery in moyamoya disease. AB - OBJECTIVE: We investigated morphologic features of the extracranial internal carotid artery (ICA) by carotid ultrasonography in patients with moyamoya disease. METHODS: We performed conventional carotid ultrasonography and transoral carotid ultrasonography on 19 ICAs in 10 patients with moyamoya disease (moyamoya group) and 28 ICAs in 14 control subjects (control group). We evaluated whether the diameter was greatly reduced at the proximal portion of the ICA above the bulbus, like a champagne bottle neck, to be less than half that of the common carotid artery and whether the diameter of the ICA was smaller than that of the external carotid artery (diameter reversal) on conventional carotid ultrasonography. We then measured the internal diameter of the extracranial distal ICA by transoral carotid ultrasonography. We compared the incidence of a "bottle neck" appearance, diameter reversal, and the ICA diameter between the 2 groups. RESULTS: The bottle neck and diameter reversal were shown in 14 (74%) and 16 (84%) of the 19 ICAs in the moyamoya group, respectively (chi2 test, P<.0001). However, neither of them was shown in the control group. The diameter of the distal ICA in the moyamoya group was significantly smaller than that in the control group (mean+/-SD, 2.4+/-0.60 versus 4.1+/-0.52 mm; unpaired t test, P<.0001). CONCLUSIONS: These results suggest that rapid internal diameter reduction at the proximal portion of the ICA, characterized by a bottle neck appearance or diameter reversal, is an important morphologic feature of moyamoya disease. PMID- 17121950 TI - Sonographically guided infraclavicular brachial plexus block in adults: a retrospective analysis of 1146 cases. AB - OBJECTIVE: The aim of this study was to analyze our experience in 1146 cases of sonographically guided infraclavicular brachial plexus block (ICBPB) performed over 32 months. METHODS: Anesthetic records of 1146 cases of sonographically guided ICBPB performed by our staff were studied retrospectively with the use of a database created by an automated anesthesia record-keeping system. The rates of successful blocks, failed blocks necessitating conversion to general anesthesia or requiring supplementation with local anesthetics, those requiring larger-than usual doses of sedation, and complications were determined. Analysis included an attempt to determine the possible causes of inadequate blocks and complications. RESULTS: In 1138 patients (99.3%), the block was successful. Six patients had incomplete blocks requiring general anesthesia, and another 2 patients needed local anesthetic supplementation by the surgeons. Ninety-seven percent of the blocks were performed by residents directly supervised by an attending anesthesiologist who held the ultrasound probe. The mean age+/-SD of the patients was 39+/-15 years; the mean duration of surgery was 165+/-114 minutes; and the male-female ratio was 4:1. More than 50% of patients were obese. There were no reported cases of nerve injury, pneumothorax, or local anesthetic toxicity. Arterial punctures occurred in 8 (0.7%) patients, but all were inconsequential. CONCLUSIONS: The data from this retrospective study suggest that sonographic guidance provides a high success rate (99.3%) and improved safety for ICBPB. The increased operator team experience virtually eliminates failure and complications. PMID- 17121951 TI - Evaluation of cardiac global function using the myocardial performance index by tissue Doppler echocardiography in patients with uremia. AB - OBJECTIVE: The purpose of this study was to assess global ventricular function in patients with uremia by means of the myocardial performance index (MPI) derived from tissue Doppler echocardiography. METHODS: According to the left ventricular mass index and pericardial effusion, 45 patients with uremia were classified into 2 groups: a uremia group and a uremia with pericardial effusion group. To calculate left ventricular MPI (LVMPI) and right ventricular MPI (RVMPI) by tissue Doppler echocardiography, the isovolumic contraction time (ICT), isovolumic relaxation time (IRT), and ejection time (ET) were measured at different sites in the mitral and tricuspid annuli. RESULTS: The mean ICT and IRT were longer, the ET was shorter, and the LVMPI and RVMPI were higher in the 2 disease groups than in a control group, and the indices were higher in the uremia with pericardial effusion group than in the uremia group. The increase of the LVMPI was more obvious than that of the RVMPI. There was a significant difference in the mean LVMPI and RVMPI among the 3 groups (P<.01). The MPI was positively correlated with the IRT and the sum of the ICT and IRT and negatively correlated with the ET. CONCLUSIONS: Both left and right ventricular systolic and diastolic function are impaired in patients with uremia. The MPI could be measured by tissue Doppler echocardiography, and we suggest that this index provides a novel, noninvasive method for clinical research on global myocardial performance in patients with uremia. PMID- 17121952 TI - Sonographically guided ilioinguinal nerve block. AB - OBJECTIVE: The aim of this study was to describe a sonographically guided ilioinguinal nerve block in adults. METHODS: We developed a useful step-by-step technique of sonographically guided ilioinguinal nerve block based on visualization of abdominal muscles, fascial planes, and the branch of the deep circumflex iliac artery. RESULTS: We performed 9 sonographic examinations with subsequent blockade of the ilioinguinal nerve. All injections resulted in a clinically successful sensory block. CONCLUSIONS: This technique is reliable and reproducible. The block is achievable by a low-volume local anesthetic injection. Visualization of the intestines and blood vessels in the abdominal wall may help prevent an inadvertent injury. PMID- 17121953 TI - Sonographic quantification of ovarian tumor vascularity. AB - OBJECTIVE: The purpose of this study was to assess the diagnostic accuracy of quantitated color Doppler sonography in differentiating benign from malignant ovarian tumors, with the use of tumor histologic examination as a reference standard. METHODS: The vascularity of 38 ovarian masses (30 benign and 8 malignant) as quantitatively depicted with color Doppler sonography was analyzed with a readily available software program (ImageJ; National Institutes of Health, Bethesda, MD). The following quantitative sonographic criteria for tumor vascularity were analyzed: the vascularity index (VI) quantified the difference between the total number of pixels and the number of pixels containing no color/totalx100, whereas the power-weighted pixel density (PWPD) weighted the strength of the signal/total. The accuracy of sonographic criteria for malignant ovarian tumors was evaluated with univariate analysis. Results of tumor histologic examination were used as proof of the final diagnosis. RESULTS: The mean values of VI and PWPD were significantly different in benign versus malignant ovarian lesions (VI, 1.3+/-1.6 versus 4.7+/-3.9; P<.01; PWPD, 2338+/ 3305 versus 9403+/-9946; P<.05). With a VI of greater than 2.3, sensitivity of 75% and specificity of 90% were obtained. When combined with a PWPD of greater than 4555, sensitivity improved to 88%, and specificity improved to 93%. Morphologic analysis had sensitivity of 72% and specificity of 76% for malignancies. CONCLUSIONS: Quantitated color Doppler sonography was found to be helpful for distinguishing benign from malignant ovarian masses. However, the wide range in values makes it most useful as an adjunct to morphologic assessment. It is anticipated that quantitated color Doppler sonography could result in a slight improvement in detection of ovarian malignancies. PMID- 17121954 TI - Variable echogenicity as a sonographic sign in the preoperative diagnosis of ovarian mucinous tumors. AB - OBJECTIVE: The purpose of this study was to evaluate the reliability of the "variable echogenicity" sign in the preoperative sonographic diagnosis of ovarian mucinous tumors. METHODS: Variable echogenicity was detected sonographically in different compartments of 8 adnexal multilocular masses. This finding was correlated with the final histologic results. RESULTS: All 8 tumors were mucinous cyst adenomas. In 3 of them, borderline malignancy was present. CONCLUSIONS: The sonographic detection of variable echogenicity in the contents of an adnexal multilocular cyst strongly suggests a mucinous tumor. PMID- 17121955 TI - Numerous intracystic floating balls as a sonographic feature of benign cystic teratoma: report of 5 cases. AB - OBJECTIVE: The purpose of this series was to describe the unusual but unique sonographic pattern of mature cystic teratoma. METHODS: Five patients of reproductive age with clinical manifestations of a pelvic mass were evaluated with sonography for treatment planning. RESULTS: All 5 cases showed the similar sonographic pattern of a large cystic tumor filled with multiple echogenic spherical structures floating in the cystic background. Of the 5 patients, 3 had rather large balls varying in size between 1 and 4 cm in the same tumor masses, whereas the other 2 had numerous smaller balls of about 0.5 cm in diameter. The numerous crowded very small echogenic balls in the last 2 cases mimicked solid nodules representing malignancy. However, there was no vascularization in the balls, which suggested a benign nature. The postoperative pathologic diagnosis was mature cystic teratoma without any malignant component in all cases. CONCLUSIONS: The sonographic feature of intracystic floating echogenic balls is probably pathognomonic for mature teratoma and is easily detected in most cases. Color Doppler sonography is helpful in differentiating these benign nodules (small balls) from malignant tumors. PMID- 17121956 TI - Pseudoaneurysm of the uterine artery after an uncomplicated spontaneous vaginal delivery. PMID- 17121957 TI - Acquired hematometra and hematotrachelos in an adolescent with dysfunctional uterine bleeding. PMID- 17121958 TI - Fracture-separation of the distal femoral epiphysis in a premature neonate. PMID- 17121959 TI - Tumorlike, iron-sparing foci of splenic hemosiderosis in a child with sickle cell anemia. PMID- 17121960 TI - Postcircumcision granuloma: a rare cause of a penile mass in a boy. PMID- 17121961 TI - Unusual appearance of a popliteal venous aneurysm in a 16-year-old patient: sonographic findings. PMID- 17121962 TI - Portal vein thrombosis may alter the correct evaluation of hepatocellular carcinoma with the sonographic contrast pulse sequence technique. PMID- 17121963 TI - Gamna-Gandy bodies: sonographic features with histopathologic correlation. PMID- 17121964 TI - Sonographic features of biliary hamartomas with histopathologic correlation. PMID- 17121966 TI - Cancer and the immortal strand hypothesis. PMID- 17121967 TI - Mapping quantitative trait loci using the experimental designs of recombinant inbred populations. AB - In the data collection of the QTL experiments using recombinant inbred (RI) populations, when individuals are genotyped for markers in a population, the trait values (phenotypes) can be obtained from the genotyped individuals (from the same population) or from some progeny of the genotyped individuals (from the different populations). Let Fu be the genotyped population and Fv (v>or=u) be the phenotyped population. The experimental designs that both marker genotypes and phenotypes are recorded on the same populations can be denoted as (Fu/Fv, u=v) designs and that genotypes and phenotypes are obtained from the different populations can be denoted as (Fu/Fv, v>u) designs. Although most of the QTL mapping experiments have been conducted on the backcross and F2(F2/F2) designs, the other (Fu/Fv, v>or=u) designs are also very popular. The great benefits of using the other (Fu/Fv, v>or=u) designs in QTL mapping include reducing cost and environmental variance by phenotyping several progeny for the genotyped individuals and taking advantages of the changes in population structures of other RI populations. Current QTL mapping methods including those for the (Fu/Fv, u=v) designs, mostly for the backcross or F2/F2 design, and for the F2/F3 design based on a one-QTL model are inadequate for the investigation of the mapping properties in the (Fu/Fv, uor=u) designs. In addition, the QTL mapping properties of the proposed and approximate methods in different designs are discussed. Simulations were performed to evaluate the performance of the proposed and approximate methods. The proposed method is proven to be able to correct the problems of the approximate and current methods for improving the resolution of genetic architecture of quantitative traits and can serve as an effective tool to explore the QTL mapping study in the system of RI populations. PMID- 17121968 TI - Characterization of metallothioneins (MT-I and MT-II) in the yak. AB - The cDNA-encoding sequences for yak metallothionein isoforms I (MT-I) and II (MT II) were amplified and cloned by reverse-transcription PCR to characterize the nucleotide sequence and protein structure of metallothionein in the yak. The cDNA sequences of MT-I and MT-II were subjected to BLAST searching at the National Center for Biotechnology Information, and the results indicated that the nucleotide sequences of yak MT-I and MT-II, when compared among different species of mammals, are highly conserved. The yak open reading frames have a length of 183 nucleotides, which encode for yak MT-I and MT-II proteins of 61 AA, respectively. Analysis of hydrophobicity, trans-membrane region, and signal peptides suggested that metallothioneins of the yak are nonsecretory proteins. There were several conserved tripeptide sequences, such as C-X-C, C-C-X-C-C, and C-X-X-C (X designates AA excluding cysteine in MT-I and MT-II), and they are highly conserved in their evolution. By homologous comparative modeling, we predicted the molecular spatial structures of yak MT-I and MT-II, which are composed of alpha- and beta-domains that are linked by the conserved tripeptide Lys(30)-Lys(31)-Ser(32) (KKS). PMID- 17121969 TI - The fatty acid composition of muscle fat and subcutaneous adipose tissue of grazing heifers supplemented with plant oil-enriched concentrates. AB - Our objective was to determine the effect of oil supplementation of pasture fed, beef cattle on the fatty acids, particularly CLA and PUFA, of muscle and s.c. adipose tissue. Forty-five Charolais crossbred heifers were blocked on BW and randomly assigned to 1 of 3 dietary regimens in a randomized complete block design (n = 15). The 3 treatments were: unsupplemented grazing (GO), restricted grazing plus a sunflower oil-enriched ration (SO), or restricted grazing plus a linseed oil-enriched ration (LO). Heifers were fed the experimental diets for approximately 158 d. Samples of LM muscle and s.c. adipose tissue were taken postmortem, the muscle fat was separated into neutral lipid and polar lipid (no separation was performed on the s.c. adipose tissue), and the fatty acid profile was determined by GLC. No effect of dietary treatment on carcass weight or total fatty acid concentration (mean 2,571 mg/100 g of muscle) in muscle fat was detected. Heifers offered SO had a greater (P < 0.001) proportion of CLA and C18:1trans-11 (1.90 and 9.35 vs. 1.35 and 6.89 g/100 g of fatty acids, respectively) in neutral lipid of muscle fat compared with those offered LO, which had a greater proportion of CLA and C18:1trans-11 than heifers offered GO (0.78 and 3.37 g/100 g of fatty acids, respectively). Similar effects were observed in the polar lipid and s.c. lipid. The PUFA:SFA ratio was greater in muscle fat and s.c. adipose tissue from supplemented heifers than in those offered GO (P < 0.001). Compared with LO, the PUFA:SFA ratio was greater (P < 0.05) in muscle fat of heifers offered SO, but there was no difference between SO and LO for this ratio in s.c. adipose tissue. The n-6:n-3 PUFA ratio was similar in muscle and s.c. adipose tissue for GO and LO, but it was greater (P < 0.05) for SO. It is concluded that supplementation of pasture-fed cattle with plant oil enriched concentrates resulted in an increase in beef fat of some fatty acids considered to be of benefit to human health. Concentrates enriched with sunflower oil were more effective in increasing the CLA concentration, whereas linseed oil enriched concentrates resulted in a more favorable n-6:n-3 PUFA ratio. The relevance to human health of the associated increase in C18:1trans-11 merits investigation. PMID- 17121970 TI - Effects of a dietary Aspergillus oryzae extract containing alpha-amylase activity on performance and carcass characteristics of finishing beef cattle. AB - Three experiments were conducted to examine the effects of an Aspergillus oryzae extract containing alpha-amylase activity on performance and carcass characteristics of finishing beef cattle. In Exp. 1, 120 crossbred steers were used in a randomized complete block design to evaluate the effects of roughage source (alfalfa hay vs. cottonseed hulls) and supplemental alpha-amylase at 950 dextrinizing units (DU)/kg of DM. Significant roughage source x alpha-amylase interactions (P < 0.05) were observed for performance. In steers fed cottonseed hulls, supplemental alpha-amylase increased ADG through d 28 and 112 and tended (P < 0.15) to increase ADG in all other periods. The increases in ADG were related to increased DMI and efficiency of gain during the initial 28-d period but were primarily related to increased DMI as the feeding period progressed. Supplemental alpha-amylase increased (P = 0.02) the LM area across both roughage sources. In Exp. 2, 96 crossbred heifers were used in a randomized complete block design with a 2 x 3 factorial arrangement of treatments to evaluate the effects of corn processing (dry cracked vs. high moisture) and supplemental alpha-amylase concentration (0, 580, or 1,160 DU/kg of DM). Alpha-amylase supplementation increased DMI (P = 0.05) and ADG (P = 0.03) during the initial 28 d on feed and carcass-adjusted ADG (P = 0.04) across corn processing methods. Longissimus muscle area was greatest (quadratic effect, P = 0.04), and yield grade was least (quadratic effect, P = 0.02) in heifers fed 580 DU of alpha-amylase/kg of DM across corn processing methods. In Exp. 3, 56 crossbred steers were used in a randomized complete block design to evaluate the effects of supplemental alpha amylase (930 DU/kg of DM) on performance when DMI was restricted to yield a programmed ADG. Alpha-amylase supplementation did not affect performance when DMI was restricted. We conclude that dietary alpha-amylase supplementation of finishing beef diets may result in increased ADG through increased DMI under certain dietary conditions and that further research is warranted to explain its mode of action and interactions with dietary ingredients. PMID- 17121971 TI - Sites of nutrient digestion in growing pigs: effect of dietary fiber. AB - The impact of dietary fiber on fecal digestion is well-known and provides a comprehensive approach toward nutrient digestibility and availability. Little quantitative information is available on digestion of fiber in the different segments of the gastrointestinal tract (GIT). The objectives of this study were to obtain a method allowing the quantification of the digestive process in different segments of the GIT and to study the impact of dietary fiber on nutrient digestibility. Six barrows (average initial BW of 30 kg and fitted with a simple T-cannula at the proximal duodenum and caudal ileum) were used in a replicated 3 x 3 Latin square design. In each period, pigs were offered 1 of 3 diets differing in fiber content (low, medium, and high). Differences in fiber content were created by replacing wheat and barley with wheat bran. Titanium dioxide was included in the diet as an indigestible marker to determine the apparent digestibility coefficients in different segments of the GIT. The apparent digestibility of ash, CP, DM, and OM increased in the different segments of the GIT. Duodenal digestibility coefficients were negative for ash (e.g., 39.9% for the medium- and high-fiber diets), indicating important endogenous mineral secretions by the stomach and digestive glands. The duodenal digestibility of other nutrients and OM were positive but close to zero and numerically lower in the diets with the greater fiber contents. The fiber content in the diet did not affect the apparent ileal digestibility of nutrients. Increasing the fiber content in the diet affected the fecal digestibility of CP, ether extract, and energy (P < 0.01). The method used for studying sites of digestion in the digestive tract provides promising results, but it is limited due to the high variability that is likely caused by sampling limitations and variation between animals. PMID- 17121972 TI - Effects of dietary n-6:n-3 fatty acid ratio on feed intake, digestibility, and fatty acid profiles of the ruminal contents, liver, and muscle of growing lambs. AB - This study investigated the effect of modifying the n-6:n-3 fatty acid ratio (FAR) of diets using linseed, soybean, and cottonseed oils on apparent digestibility, ruminal fermentation characteristics, growth performance, key circulating hormones, and the fatty acid profile of ruminal digesta, liver, and fore-shank muscle of growing lambs fed a high concentrate diet. Forty individually housed Katadhin Dorper lambs (average of 20.0 kg of BW) were fed Bermudagrass hay in ad libitum amounts and concentrates at 3.7% of BW daily. The concentrate contained 68.9% corn, 23.8% soybean meal, 3.3% limestone, and 4.0% oil supplements (DM basis). The treatments consisted of dietary n-6:n-3 FAR of 2.3:1, 8.8:1, 12.8:1, and 15.6:1. After feeding for 35 d in metabolism crates, lambs were slaughtered 15 h after feeding, and samples of ruminal digesta, blood, liver, and foreshank tissue were collected. Increasing dietary n-6:n-3 FAR did not affect the intake of DM nor the apparent digestibility of DM, ether extract, NDF, or ADF, but did increase apparent digestibility of CP (linear, P < 0.05). Concentrations of ruminal butyrate increased linearly (P < 0.05) with increasing dietary n-6:n-3 FAR, whereas the valerate concentration decreased linearly (P < 0.001). Concentrations of plasma insulin and IGF-I were not affected by dietary n 6:n-3 FAR. Concentrations of C18:3n-3 increased linearly (P < 0.001), whereas that of C18:2n-6 decreased linearly (P < 0.001) in ruminal digesta with decreasing dietary n-6:n-3 FAR. Concentrations of transisomers of fatty acids in ruminal digesta did not change. Proportions of C18:0 in liver and foreshank muscle were unchanged by diet. The proportion of trans11 C18:1 and cis-9 trans11 CLA decreased (P < 0.05) in liver but increased (P < 0.05) in foreshank muscle as dietary n-6:n-3 FAR decreased. Proportions of all measured n-3 fatty acids were greater in liver when diets contained more C18:3n-3 from linseed oil. By decreasing the dietary n-6:n-3 FAR, the proportions of n-6 fatty acids in foreshank muscle decreased dramatically; specifically, C18:2n-6 decreased linearly (P < 0.001) from 28.0 to 16.5% and C20:4n-6 decreased linearly (P < 0.001) from 14.7 to 8.6%. Although feeding a diet that contained more n-3 fatty acids increased the n-3 fatty acid concentration of muscle, the ratio of PUFA to SFA was decreased. PMID- 17121973 TI - Haplotype analysis of the DQA genes in sheep: evidence supporting recombination between the loci. AB - The ovine class II major histocompatibility complex mediates specific immune responses to exogenous antigens in sheep. A number of ovine class II loci have been identified, and most of them appear to be polymorphic. In this study we investigated the DQA1 locus of 520 sheep and the DQA2 locus of over 40,000 sheep, finding 12 sequences and 22 sequences, respectively, using DQA1- and DQA2 specific PCR primers. Among the DQA2 sequences, 2 groups of sequences can be found: those that share homology with the DQA2 sequences from closely related species and those that cluster with bovine DQA3 and DQA4 sequences and have been called DQA2-like in sheep. The occurrence of these DQA2-like sequences was once again confirmed to correspond with the absence of detectable DQA1 sequences, suggesting that they are found at the same location as DQA1. Within the sheep studied, 37 haplotypes could be detected, 23 being haplotypes of DQA1 and DQA2 sequences and with frequencies ranging from 0.38 to 9.27%, and 14 being haplotypes of DQA2 and DQA2-like sequences and with frequencies ranging from 0.03 to 14.53%. We discovered 12 DQA1-DQA2 combinations that were derived from 5 DQA1 alleles and 4 DQA2 alleles, and 8 DQA2-DQA2-like combinations from 5 DQA2 alleles and 2 DQA2-like sequences. The frequency of occurrence of recombined DQA1-DQA2 sequences and recombined DQA2-DQA2-like sequences is similar, once again suggesting the DQA2-like sequences are found at the DQA1 locus. PMID- 17121974 TI - Effects of added fermentable carbohydrates in the diet on intestinal proinflammatory cytokine-specific mRNA content in weaning piglets. AB - There is increasing evidence showing that dietary supplementation with prebiotics can be effective in the treatment of intestinal inflammation. Because weaning time is characterized by rapid intestinal inflammation, this study investigated the effect of a diet supplemented with a combination of 4 fermentable carbohydrates (lactulose, inulin, sugarbeet pulp, and wheat starch) on the mRNA content of proinflammatory cytokines in newly weaned piglets. Cytokines (IL 1beta, IL-6, IL-8, IL-12p40, IL-18, and tumor necrosis factor-alpha) were analyzed using a semiquantitative reverse-transcription PCR technique on d 1, 4, and 10 in the ileum and colon of piglets fed either a test diet (CHO) or a control diet. In addition to the diet, the effect of enforced fasting on cytokine mRNA content was also evaluated. No effect of fasting was observed on the pro inflammatory cytokine mRNA content. Our results showed that the CHO diet induced an up-regulation of IL-6 mRNA content in the colon of piglets 4 d postweaning. This up-regulation was specific for the animals fed the CHO diet and was not observed in animals fed the control diet. An increase in IL-1beta mRNA content was also observed on d 4 postweaning in all of the piglets. Correlations between proinflammatory cytokines and the end-products of fermentation indicated that the regulation of cytokines may be linked with some of the fermentation end-products such as branched-chain fatty acids, which are in turn end-products of protein fermentation. PMID- 17121975 TI - The effect of feeding low-phytate barley-soybean meal diets differing in protein content to growing pigs on the excretion of phosphorus and nitrogen. AB - An experiment was conducted with growing pigs to determine the excretion of P and N in 4 barley-based diets formulated to contain 18 or 15% CP by using a normal barley (NB) or a low-phytate barley (LPB). The NB contained 0.31% total P and 0.19% phytate P; the LPB contained 0.32% total P and 0.01% phytate P. The diets were supplemented, when so required, with lysine, methionine, threonine, and tryptophan to meet their apparent ileal digestible supplies according to the NRC (1998). The diets containing NB were supplemented with inorganic P to meet the NRC (1998) recommendation for available P (0.23%). The diets containing LPB were not supplemented with inorganic P because these contained sufficient available P (0.27%). Eight barrows with an average BW of 20.9 kg were assigned to the 4 dietary treatments according to a repeated 4 x 4 Latin square design. The diets were fed at a rate of 2.5 times the ME requirement for maintenance. The barrows were fed twice daily, at 0800 and 1500, equal amounts each meal. Water was added to the feed at a ratio of 2.5:1. Each experimental period consisted of a 7-d adaptation period followed by a 5-d collection of feces and urine. The substitution of NB with LPB decreased (P < 0.001) the total P excretion by 38 and 43% for the 18 and 15% CP diets, respectively. Reducing the CP content from 18 to 15% decreased (P < 0.001) the N excretion by 29 and 32% for the NB and LPB diets, respectively. With the reduction in CP content, there was a decrease (P < 0.001) in the amount of N retained. The N:P ratio in manure of pigs fed the LPB diets was greater (P < 0.001) than from pigs fed the NB diets. These data indicate that P and N excretion can be greatly reduced by substitution of NB by LPB, and also by the reduction of the CP content, in diets for growing pigs. PMID- 17121976 TI - Choice of attractive conditions by beef cattle in a Y-maze just after release from restraint. AB - To provide useful information on how to moderate posthandling stress, the attractiveness of different conditions to beef cattle just after release from restraint was determined. Angus heifers were individually allowed to enter a choice area after 2 min of restraint in a squeeze chute and to choose between 2 pens. After the heifer had chosen a pen, it could freely access both test pens and the choice area for a further 5 min. In Exp. 1, each heifer was given 1 of the following choices: a pen with 3 familiar heifers (PEERS) vs. a pen with a pile of hay on a metal rack (FOOD; n = 34); PEERS vs. the bare pen (BARE; n = 34); and FOOD vs. BARE (n = 35). When the choice combination was PEERS vs. BARE, more heifers chose PEERS (P < 0.05). When the choice combination was PEERS vs. FOOD, more heifers than expected tended to choose PEERS (P < 0.10), whereas FOOD and BARE did not differ. The latency to choose either pen was shorter if PEERS was 1 of the 2 choices (P < 0.01). After choosing, more heifers entered the PEERS pen than the FOOD (P < 0.05) or BARE (P < 0.01) pens. In Exp. 2, another 86 heifers were given 1 of the following choices: a pen with a familiar handler standing inside (STI) vs. a pen with a novel object (NO; n = 29); a pen with the handler standing outside the pen (STO) vs. NO (n = 29); a pen in which the handler was sitting inside (SI) vs. NO (n = 28). Fewer heifers chose the pen with the human (STI, STO, and SI; all P < 0.01). Except for the choice of STO vs. NO, the number of heifers that had voluntarily chosen either pen was larger than expected (STI and SI; both P < 0.01). The number of times in which the NO pen was entered was greater than the STI and STO (P < 0.01), although the number of times in which the SI and NO pens were entered was not different. More heifers avoided the human, particularly a standing human. In conclusion, just after handling with restraint, returning cattle to the group of peers and not approaching the cattle needlessly should moderate their stress. PMID- 17121977 TI - Steroid hormone secretion during the ovulatory cycle and pregnancy in farmed Alaskan reindeer. AB - Seasonal endocrine changes in 5 non-bred and 10 pregnant Alaskan reindeer have been documented. Blood samples were collected from early September until early May, spanning the breeding season, gestation, or the anovulatory period. Plasma was analyzed by RIA for progesterone (P4), estradiol-17beta, estrone, and estrone sulfate. Elevated P4 in 80% of the reindeer at the onset of the study indicated that ovarian activity had been initiated. The median date for the onset of the first recorded full-length ovulatory cycle was September 23. In nonbred reindeer, the mean ovulatory cycle length from September to May was 24 +/- 1 d (range 18 to 29 d). Nonbred females continued to cycle throughout the winter, displaying 6 to 8 ovulatory cycles after the beginning of blood sampling. Cycle length (mean 22 to 24 d) did not vary between individuals (P = 0.170) or over the course of the winter (P = 0.244). In early April, ovulatory cycles ceased with normal demise of the corpus luteum in 2 females, whereas the remaining 3 females formed apparently persistent corpora lutea. Natural service breeding occurred between September 10 and October 2, and P4 profiles indicated that all breeding females conceived to the first mating. Concentrations of P4 rose steadily after conception and remained elevated throughout gestation, with mean concentrations not varying significantly (P = 0.104) from 4 to 28 wk of gestation. Estrogens all followed patterns similar to each other, remaining at baseline concentrations until approximately 24 wk of gestation and rising coincidently as P4 declined just before parturition. There was a continual overlap throughout the winter in peak P4 concentrations observed in cycling and pregnant reindeer. Calving occurred between April 8 and May 2, resulting in a mean gestation length of 211 +/- 2.2 d (range 198 to 221 d). Information from this study can be used by Alaskan reindeer producers to improve management and profitability of reindeer production. PMID- 17121978 TI - The CHRNE 470del20 mutation causing congenital myasthenic syndrome in South African Brahman cattle: prevalence, origin, and association with performance traits. AB - Genotyping of the South African, registered, Brahman cattle population for the 470del20 mutation in the CHRNE gene causing congenital myasthenic syndrome (CMS) was carried out in 1,453 animals. Overall prevalence of carriers was 0.97% (0.50 to 1.68%, 95% confidence interval). Carrier prevalence among breeding bulls in 2004 was 1.22% (0.65 to 2.15%, 95% confidence interval), and had not changed significantly since 2000. Using segregation analysis, CMS genotype probabilities were calculated for all 612,219 animals in the pedigree, leading to the identification of 2 founder animals as the most likely original carriers. Pedigree analysis revealed no ancestors common to all known carriers, but rather that the mutation had been introduced at least twice into the South African Brahman population, probably via animals imported from the United States. The effects of CMS genotype probability on adjusted birth, 200-d, 400-d, and 600-d BW, as well as on EBV for birth, 200-d, 400-d, and 600-d BW, and milk, were estimated, accounting for effects of sire. Heterozygosity for the CHRNE 470del20 mutation was associated with a 13.3-kg increase in adjusted 600-d BW (P = 0.03). Positive effects of CMS carrier status on all BW EBV were found, but no effect was found on milk EBV. We conclude that CMS carriers have a BW advantage at 600 d and possibly also at birth, 200 d, and 400 d. This may confer a selective advantage and tend to increase the frequency of the mutation. PMID- 17121979 TI - Influence of lipid content on pork sensory quality within pH classification. AB - The objective of this project was to determine the contribution of lipid content to textural and sensory properties of fresh pork within defined pH classifications. Pigs (n = 1,535; from 248 sires and 836 dams) from the 1991, 1992, and 1994 National Barrow Show Sire Progeny Test were used in this study. The test included purebred Berkshire (107), Chester White (113), Duroc (249), Hampshire (220), Landrace (165), Poland China (101), Spotted (181), and Yorkshire (399) barrows (901) and gilts (634). Diets were uniform across breeds within test. The halothane (Hal 1843) genotype (1346 NN and 189 Nn) was determined. Pigs were slaughtered at 105 kg of BW, and samples of the LM were obtained from each carcass at the 10th rib. Star probe, sensory traits, and lipid content were determined on the LM from each pig. A pH classification of LM was assigned as follows: class A, > 5.95, n = 186; class B, > or = 5.80 to 5.95, n = 236; class C, > or = 5.65 to 5.80, n = 467; class D, > or = 5.50 to 5.65, n = 441; class E, < 5.50, n = 205. Data were analyzed using a mixed linear model including pH classification, test, sex, halothane genotype, breed, and breed x sex interaction as fixed effects, with sire and dam within breed included as random effects. Correlations were determined within pH class. Lipid content was a significant source of variation for models predicting star probe values in class C and D and for chewiness in class B, C, and D. Increasing lipid content tended to increase sensory tenderness in pH class D. Sensory tenderness was not affected by lipid content in pH class A, B, or E. Lipid content was not a significant source of variation for juiciness scores within any pH class. Intramuscular lipid is correlated with sensory texture traits primarily in classes C and D. Within class C and D, correlations indicate that increasing lipid content is associated with high sensory tenderness, low sensory chewiness, and low star probe values. It is concluded that lipid content is a small source of variation in texture and tenderness of pork loin with pH between 5.80 and 5.50, but not at a greater or lesser pH. PMID- 17121980 TI - Technical note: A model to estimate individual feed intake of swine in group feeding. AB - In most animal growth experiments, groups of animals are housed within a pen. Occasionally, an individual animal shows a very different growth rate than its pen mates or dies during the experiment. When this happens, if pen feed intake (PFI) cannot be reestimated for the calculation of ADFI and feed efficiency, an observation will be lost from the data set. Therefore, we propose a model to estimate individual feed intake (IFI) of pigs in group feeding, with subsequent validation of the model using group feeding simulation studies. In the proposed model, the feed intake (FI) of each affected pen is partitioned into FI for maintenance (FI(m)) and FI for growth (FI(g)) for each animal within that pen. First, individual pig FI(m) for the period is calculated using the 1998 National Research Council estimation of ME for maintenance. Then, FI(m) for all pigs in the pen is summed. The difference between the summed FI(m) and the total PFI is that which supported growth in the pen. Next, FI(g) is calculated by apportioning the remaining feed equally to each unit of gain within the pen. Finally, the estimated IFI for the pig being removed from the pen is the sum of FI(m) and FI(g) for that pig; this FI estimate is subtracted from the original PFI to leave the new PFI for the remaining pigs. The validity of the estimated IFI is dependent on the accuracy of the maintenance energy equation and the energy analysis of the feedstuffs. In simulation studies, we compared the accuracy of the proposed method with 2 other methods. In simulation study 1, the proposed model showed better accuracy than at least one of the other methods during all tested periods (P < 0.001). In simulation study 2, the greater accuracy of the proposed method compared with 2 other methods was demonstrated again. Because calculation of IFI is relatively cumbersome, we developed a feed intake correction spreadsheet (FICS), an Excel spreadsheet containing macros for FI correction. All of the calculation procedures in the proposed model are included within the feed intake correction spreadsheet. The Excel file and instructions are being made available via a Web site. PMID- 17121981 TI - Preparation of monoclonal antibodies for detection and identification of Francisella tularensis. AB - Monoclonal antibodies (MAbs) against Francisella tularensis were obtained. Three MAbs specifically reacted with F. tularensis, while four MAbs reacted with other members of the genus Francisella as well. Fluorescent isothiocyanate-conjugated MAbs unequivocally stained bacterial cells in specimens from experimentally infected mice. Two MAbs agglutinated F. tularensis antigen in the agglutination tests. These MAbs should improve methods for detection and identification of F. tularensis. PMID- 17121982 TI - Performance of a commercial immunoglobulin M antibody capture assay using analyte specific reagents to screen for interfering factors during a West Nile virus epidemic season in Nebraska. AB - In 2003, the Nebraska Public Health Laboratory tested more than 10,371 serum and 516 cerebral spinal fluid specimens. Results showed that without performing the interfering factors screen for specimens in the low positive index value range of >1.1 to 20%. Plasma carotenoids were in turn responsible for individual differences in coloration and immune response. Our results provide experimental evidence for a link between testosterone levels and immunoenhancing carotenoids that (i) reconciles conflicting evidence for the immunosuppressive nature of androgens, (ii) provides physiological grounds for a connection between two of the main signaling systems in animals, (iii) explains how these signaling systems can be evolutionary stable and honest, and (iv) may explain the high prevalence of sexual dimorphism in carotenoid-based coloration in animals. PMID- 17121985 TI - Activation of an oncogenic microRNA cistron by provirus integration. AB - Retroviruses can cause tumors when they integrate near a protooncogene or tumor suppressor gene of the host. We infected >2,500 mice with the SL3-3 murine leukemia virus; in 22 resulting tumors, we found provirus integrations nearby or within the gene that contains the mir-17-92 microRNA (miRNA) cistron. Using quantitative real-time PCR, we showed that expression of miRNA was increased in these tumors, indicating that retroviral infection can induce expression of oncogenic miRNAs. Our results demonstrate that retroviral mutagenesis can be a potent tool for miRNA discovery. PMID- 17121986 TI - Rapid and reversible changes in intrahippocampal connectivity during the course of hibernation in European hamsters. AB - The hippocampal formation is a highly plastic brain structure that undergoes structural remodeling in response to internal and external challenges such as metabolic imbalance and repeated stress. We investigated whether the extreme alterations in metabolic status that occur during the course of hibernation in European hamsters cause structural changes in the dendritic arborizations of the CA3 pyramidal neurons and their main excitatory afferents, the mossy fiber terminals (MFT), that originate in the dentate gyrus. We report that apical, but not basal, dendritic trees of Golgi-impregnated CA3 principal neurons are significantly shorter, less branched, and less spiny in hypothermic hamsters compared with active animals. After the induction of arousal from torpor, within 2 h, the apical dendritic lengths, branching patterns, and spine density estimations returned to levels found in active, euthermic hamsters. The ultrastructure of MFT in hibernating hamsters showed a significant reduction in synaptic vesicle density and in the percentage of MFT area covered by spine profiles. Awakened hamsters showed restoration of MFT morphology to that seen in active animals. MFT of torpid animals also showed a significant increase in the percentage area of mitochondrial profiles that remained higher 3 h after induced arousal from hibernation compared with euthermic controls. Thus, the torpid/awakening cycle of the hibernating European hamster causes a rapid and reversible morphological reorganization of intrahippocampal subregions involved in information processing. The reported reductions in morphological connectivity between the dentate gyrus and the CA3 subregions could underlie the cessation of exploratory activity and spatial navigation skills during hibernation. PMID- 17121987 TI - Multiple valence superatoms. AB - We recently demonstrated that, in gas phase clusters containing aluminum and iodine atoms, an Al(13) cluster behaves like a halogen atom, whereas an Al(14) cluster exhibits properties analogous to an alkaline earth atom. These observations, together with our findings that Al(13)(-) is inert like a rare gas atom, have reinforced the idea that chosen clusters can exhibit chemical behaviors reminiscent of atoms in the periodic table, offering the exciting prospect of a new dimension of the periodic table formed by cluster elements, called superatoms. As the behavior of clusters can be controlled by size and composition, the superatoms offer the potential to create unique compounds with tailored properties. In this article, we provide evidence of an additional class of superatoms, namely Al(7)(-), that exhibit multiple valences, like some of the elements in the periodic table, and hence have the potential to form stable compounds when combined with other atoms. These findings support the contention that there should be no limitation in finding clusters, which mimic virtually all members of the periodic table. PMID- 17121988 TI - Continuous tooth generation in mouse is induced by activated epithelial Wnt/beta catenin signaling. AB - The single replacement from milk teeth to permanent teeth makes mammalian teeth different from teeth of most nonmammalian vertebrates and other epithelial organs such as hair and feathers, whose continuous replacement has been linked to Wnt signaling. Here we show that mouse tooth buds expressing stabilized beta-catenin in epithelium give rise to dozens of teeth. The molar crowns, however, are typically simplified unicusped cones. We demonstrate that the supernumerary teeth develop by a renewal process where new signaling centers, the enamel knots, bud off from the existing dental epithelium. The basic aspects of the unlocked tooth renewal can be reproduced with a computer model on tooth development by increasing the intrinsic level of activator production, supporting the role of beta-catenin pathway as an upstream activator of enamel knot formation. These results may implicate Wnt signaling in tooth renewal, a capacity that was all but lost when mammals evolved progressively more complicated tooth shapes. PMID- 17121989 TI - Autoantibody-mediated demyelination depends on complement activation but not activatory Fc-receptors. AB - The precise mechanisms leading to CNS inflammation and myelin destruction in both multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE) remain the subject of intense debate. In both MS and EAE, autoantibodies (autoAbs) are thought to be involved in tissue destruction through recruiting Fc receptor (FcR)-bearing cells or direct cytotoxic effects through the activation of the complement pathway. Whereas intrathecal immunoglobulin (Ig) production and Ig deposition in inflammatory lesions is a hallmark of MS, mice deficient in B cells and Igs develop severe EAE. Paradoxically, mice of the same genetic background but deficient in FcRgamma are EAE-resistant. We found that the functional expression of FcRgamma on systemic accessory cells, but not CNS resident cells, appears to be vital for the development of CNS inflammation, independent of antigen-presenting cell function or Ab involvement. On the other hand, we found that the injection of antimyelin oligodendrocyte glycoprotein-Abs drastically worsens disease severity, inflammation, and demyelination. Using FcRgamma(-/-) and C1q(-/-) mice, we could definitively establish that the demyelinating capacity of such autoAb in vivo relies entirely on complement activation and is FcR-independent. PMID- 17121990 TI - Ameiotic recombination in asexual lineages of Daphnia. AB - Despite the enormous theoretical attention given to the evolutionary consequences of sexual reproduction, the validity of the key assumptions on which the theory depends rarely has been evaluated. It is often argued that a reduced ability to purge deleterious mutations condemns asexual lineages to an early extinction. However, most well characterized asexual lineages fail to exhibit the high levels of neutral allelic divergence expected in the absence of recombination. With purely descriptive data, it is difficult to evaluate whether this pattern is a consequence of the rapid demise of asexual lineages, an unusual degree of mutational stability, or recombination. Here, we show in mutation-accumulation lines of asexual Daphnia that the rate of loss of nucleotide heterozygosity by ameiotic recombination is substantially greater than the rate of introduction of new variation by mutation. This suggests that the evolutionary potential of asexual diploid species is not only a matter of mutation accumulation and reduced efficiency of selection, and [corrected] it underscores the limited utility of using neutral allelic divergence as an indicator of ancient asexuality. PMID- 17121991 TI - Hypoxia facilitates Alzheimer's disease pathogenesis by up-regulating BACE1 gene expression. AB - The molecular mechanism underlying the pathogenesis of the majority of cases of sporadic Alzheimer's disease (AD) is unknown. A history of stroke was found to be associated with development of some AD cases, especially in the presence of vascular risk factors. Reduced cerebral perfusion is a common vascular component among AD risk factors, and hypoxia is a direct consequence of hypoperfusion. Previously we showed that expression of the beta-site beta-amyloid precursor protein (APP) cleavage enzyme 1 (BACE1) gene BACE1 is tightly controlled at both the transcriptional and translational levels and that increased BACE1 maturation contributes to the AD pathogenesis in Down's syndrome. Here we have identified a functional hypoxia-responsive element in the BACE1 gene promoter. Hypoxia up regulated beta-secretase cleavage of APP and amyloid-beta protein (Abeta) production by increasing BACE1 gene transcription and expression both in vitro and in vivo. Hypoxia treatment markedly increased Abeta deposition and neuritic plaque formation and potentiated the memory deficit in Swedish mutant APP transgenic mice. Taken together, our results clearly demonstrate that hypoxia can facilitate AD pathogenesis, and they provide a molecular mechanism linking vascular factors to AD. Our study suggests that interventions to improve cerebral perfusion may benefit AD patients. PMID- 17121992 TI - The dynamics behind Titan's methane clouds. AB - We present results of an axisymmetric global circulation model of Titan with a simplified suite of atmospheric physics forced by seasonally varying insolation. The recent discovery of midlatitude tropospheric clouds on Titan has caused much excitement about the roles of surface sources of methane and the global circulation in forming clouds. Although localized surface sources, such as methane geysers or "cryovolcanoes," have been invoked to explain these clouds, we find in this work that clouds appear in regions of convergence by the mean meridional circulation and over the poles during solstices, where the solar forcing reaches its seasonal maximum. Other regions are inhibited from forming clouds because of dynamical transports of methane and strong subsidence. We find that for a variety of moist regimes, i.e., with the effect of methane thermodynamics included, the observed cloud features can be explained by the large-scale dynamics of the atmosphere. Clouds at the solsticial pole are found to be a robust feature of Titan's dynamics, whereas isolated midlatitude clouds are present exclusively in a variety of moist dynamical regimes. In all cases, even without including methane thermodynamics, our model ceases to produce polar clouds approximately 4-6 terrestrial years after solstices. PMID- 17121993 TI - PKB and megalin determine the survival or death of renal proximal tubule cells. AB - Renal proximal tubule cells have a remarkable ability to reabsorb large quantities of albumin through megalin-mediated endocytosis. This is an essential process for overall body homeostasis. Overstressing this endocytic system with a prolonged excess of albumin is injurious to proximal tubule cells. How these cells function and protect themselves from injury is unknown. Here, we show that megalin is the sensor that determines whether cells will be protected or injured by albumin. Megalin, through a novel mechanism, binds PKB in a D-3-phosphorylated phospholipid-insensitive manner, anchoring PKB in the luminal plasma membrane. Whereas low doses of albumin are protective, an overload of albumin decreases megalin expression followed by a reduction of plasma membrane PKB, PKB activity, and Bad phosphorylation induced by PKB. The result is albumin-induced apoptosis. These results reveal a model for PKB distribution in the plasma membrane and elucidate mechanisms involved in both the protective and toxic effects of albumin on proximal tubule cells. In addition, our findings suggest a mechanism for the progression of chronic kidney disease to end-stage renal disease. PMID- 17121994 TI - The promyelocytic leukemia protein functions as a negative regulator of IFN-gamma signaling. AB - IFN-gamma is an immunomodulatory cytokine and uses the STAT-1alpha transcription factor to mediate gene expression. The promyelocytic leukemia (PML) protein regulates transcription as an activator or repressor, depending on the gene under investigation. Herein, we examined the influence of PML on IFN-gamma signaling, using PML wild-type (Pml(+/+)) and deficient (Pml(-/-)) mouse embryonic fibroblasts (MEF). Pml(-/-) MEF exhibit enhanced IFN-gamma-induced STAT-1alpha transcriptional activity compared with Pml(+/+) cells. Moreover, reconstitution of PML in Pml(-/-) MEF reduced STAT-1alpha transcriptional activity to levels comparable to Pml(+/+) MEF. Numerous endogenous IFN-gamma-regulated genes were up regulated in Pml(-/-) MEF compared with Pml(+/+) MEF. IFN-gamma-mediated STAT 1alpha DNA-binding activity was enhanced in Pml(-/-) cells compared with Pml(+/+) cells. Lastly, IFN-gamma enhanced the formation of a PML-STAT-1alpha complex in the nucleus. These data suggest a novel function for PML in the IFN-gamma signaling pathway by inhibiting STAT-1alpha DNA binding and transcriptional activity. PMID- 17121995 TI - Reconstructing repressor protein levels from expression of gene targets in Escherichia coli. AB - The basic underlying problem in reverse engineering of gene regulatory networks from gene expression data is that the expression of a gene encoding the regulator provides only limited information about its protein activity. The proteins, which result from translation, are subject to stringent posttranscriptional control and modification. Often, it is only the modified version of the protein that is capable of activating or repressing its regulatory targets. At present there exists no reliable high-throughput technology to measure the protein activity levels in real-time, and therefore they are, so-to-say, lost in translation. However, these activity levels can be recovered by studying the gene expression of their targets. Here, we describe a computational approach to predict temporal regulator activity levels from the gene expression of its transcriptional targets in a network motif with one regulator and many targets. We consider an example of an SOS repair system, and computationally infer the regulator activity of its master repressor, LexA. The reconstructed activity profile of LexA exhibits a behavior that is similar to the experimentally measured profile of this repressor: after UV irradiation, the amount of LexA substantially decreases within a few minutes, followed by a recovery to its normal level. Our approach can easily be applied to known single-input motifs in other organisms. PMID- 17121996 TI - Inference for nonlinear dynamical systems. AB - Nonlinear stochastic dynamical systems are widely used to model systems across the sciences and engineering. Such models are natural to formulate and can be analyzed mathematically and numerically. However, difficulties associated with inference from time-series data about unknown parameters in these models have been a constraint on their application. We present a new method that makes maximum likelihood estimation feasible for partially-observed nonlinear stochastic dynamical systems (also known as state-space models) where this was not previously the case. The method is based on a sequence of filtering operations which are shown to converge to a maximum likelihood parameter estimate. We make use of recent advances in nonlinear filtering in the implementation of the algorithm. We apply the method to the study of cholera in Bangladesh. We construct confidence intervals, perform residual analysis, and apply other diagnostics. Our analysis, based upon a model capturing the intrinsic nonlinear dynamics of the system, reveals some effects overlooked by previous studies. PMID- 17121997 TI - A structural model reveals energy transduction in dynein. AB - Intracellular active transport is driven by ATP-hydrolyzing motor proteins that move along cytoskeletal filaments. In particular, the microtubule-associated dynein motor is involved in the transport of organelles and vesicles, the maintenance of the Golgi, and mitosis. However, unlike kinesin and myosin, the mechanism by which dynein converts chemical energy into mechanical force remains largely a mystery, due primarily to the lack of a high-resolution molecular structure. Using homology modeling and normal mode analysis, we propose a complete atomic structure and a mechanism for force generation by the motor protein dynein. In agreement with very recent electron microscopy (EM) reconstructions showing dynein as a ring-shaped heptamer, our model consists of six ATPases of the AAA (ATPases associated with various cellular activities) superfamily and a C-terminal domain, which is experimentally known to control motor function. Our model shows a coiled coil spanning the diameter of the motor that accounts for previously unidentified structures in EM studies and provides a potential mechanism for long-range communication between the AAA domains. Furthermore, normal mode analysis reveals that the subunits of the motor that contain the nucleotide binding sites exhibit minimal movement, whereas the rest of the motor is very mobile. Our analysis suggests the likely domain rearrangements of the motor unit that generate its power stroke. This study provides insights into the structure and function of dynein that can guide further experimental investigations into energy transduction in dynein. PMID- 17121998 TI - PCR-based diagnosis of Naegleria sp. infection in formalin-fixed and paraffin embedded brain sections. AB - We developed a real-time PCR which allowed the highly sensitive detection of Naegleria fowleri in histological brain tissue sections from experimentally infected mice. This genus-specific small-subunit (18S) rRNA gene-based PCR can complement conventional (immuno-) histology for the diagnosis of primary amoebic meningoencephalitis in paraffin-embedded brain necropsy specimens that had been fixed in formalin buffered with phosphate-buffered saline. PMID- 17121999 TI - A multiplex ligase detection reaction-fluorescent microsphere assay for simultaneous detection of single nucleotide polymorphisms associated with Plasmodium falciparum drug resistance. AB - Incomplete malaria control efforts have resulted in a worldwide increase in resistance to drugs used to treat the disease. A complex array of mutations underlying antimalarial drug resistance complicates efficient monitoring of parasite populations and limits the success of malaria control efforts in regions of endemicity. To improve the surveillance of Plasmodium falciparum drug resistance, we developed a multiplex ligase detection reaction-fluorescent microsphere-based assay (LDR-FMA) that identifies single nucleotide polymorphisms (SNPs) in the P. falciparum dhfr (9 alleles), dhps (10 alleles), and pfcrt (3 alleles) genes associated with resistance to Fansidar and chloroquine. We evaluated 1,121 blood samples from study participants in the Wosera region of Papua New Guinea, where malaria is endemic. Results showed that 468 samples were P. falciparum negative and 453 samples were P. falciparum positive by a Plasmodium species assay and all three gene assays (concordance, 82.2%). For P. falciparum infections where the assay for each gene was positive, 2 samples carried resistance alleles for all three genes, 299 carried resistance alleles for dhfr and pfcrt, 131 carried resistance alleles for only one gene (dhfr [n = 40], dhps [n = 1], or pfcrt [n = 90]), and 21 carried only sensitive alleles at all three genes. Mixed-strain infections characterized 100 samples. Overall, 95.4% (432/453) of P. falciparum-infected samples carried at least one allele associated with resistance to Fansidar or chloroquine. In view of the fact that 86.3% (391/453) of P. falciparum-infected samples carried pfcrt mutations, chloroquine is largely ineffective against P. falciparum in Papua New Guinea. Surveillance of additional dhfr and dhps polymorphisms in order to monitor the continued effectiveness of Fansidar is recommended. PMID- 17122000 TI - Development and clinical application of a panfungal PCR assay to detect and identify fungal DNA in tissue specimens. AB - Given the rise in the incidence of invasive fungal infections (IFIs) and the expanding spectrum of fungal pathogens, early and accurate identification of the causative pathogen is essential. We developed a panfungal PCR assay that targets the internal transcribed spacer 1 (ITS1) region of the ribosomal DNA gene cluster to detect fungal DNA in fresh and formalin-fixed, paraffin-embedded (PE) tissue specimens from patients with culture-proven (n=38) or solely histologically proven (n=24) IFIs. PCR products were sequenced and compared with sequences in the GenBank database to identify the causal pathogen. The molecular identification was correlated with results from histological examination and culture. The assay successfully detected and identified the fungal pathogen in 93.6% and 64.3% of culture-proven and solely histologically proven cases of IFI, respectively. A diverse range of fungal genera were identified, including species of Candida, Cryptococcus, Trichosporon, Aspergillus, Fusarium, Scedosporium, Exophiala, Exserohilum, Apophysomyces, Actinomucor, and Rhizopus. For five specimens, molecular analysis identified a pathogen closely related to that identified by culture. All PCR-negative specimens (n=10) were PE tissues in which fungal hyphae were visualized. The results support the use of the panfungal PCR assay in combination with conventional laboratory tests for accurate identification of fungi in tissue specimens. PMID- 17122001 TI - First reported infections caused by three newly described genera in the family Xanthomonadaceae. AB - Members of the family of Xanthomonadaceae are typically characterized as environmental organisms. With the exception of Stenotrophomonas maltophilia, these organisms are infrequently implicated as human pathogens. We describe three cases of central venous catheter-associated bloodstream infections caused by Dokdonella koreensis, Aquimonas voraii, and a Luteibacter sp., all newly named genera within the family Xanthomonadaceae. The three patients all had histories of underlying hematological disorders, presented with fever, and recovered fully following treatment. These isolates required 16S rRNA gene sequencing for identification and, unlike S. maltophilia, demonstrated susceptibility to most antibiotics tested. This report represents the first description of human infections caused by these organisms. PMID- 17122002 TI - Molecular epidemiology of metallo-beta-lactamase-producing Pseudomonas aeruginosa in the Calgary Health Region: emergence of VIM-2-producing isolates. AB - A study was designed to describe the molecular epidemiology of carbapenem resistant (CR) Pseudomonas aeruginosa in a large well-defined geographical region with a centralized laboratory system serving one pediatric and three large adult hospitals (acute care centers I, II, and III). Molecular characterization was done using PCR with sequencing of the integron-associated gene cassettes. Pulsed field gel electrophoresis (PFGE) using a modified combined Stenotrophomas maltophilia and Streptococcus pneumoniae protocol with SpeI was performed on CR P. aeruginosa strains isolated in the Calgary Health Region during 2002-2006. The majority (96%) of metallo-beta-lactamase (MBL)-producing isolates produced VIM-2 with gene cassettes aacC1 and aacA4, while 4% produced IMP-7 with gene cassettes aacC4 and aacC1. Eighty-six percent of VIM-2 producers belonged to a cluster (MBLV) that was responsible for nosocomial outbreaks during 2003 (intensive care unit) and 2004 (bone marrow transplant unit) at acute care center I. Environmental isolates from these units also belonged to MBLV. The majority of strains from cluster MBLVR (related to MBLV) were present in acute care center III. Isolates producing IMP-7 belonged to a different cluster (MBLI) and were related to strains described during the 1990 s. PFGE of the MBL-negative CR strains showed that 37% belonged to a closely related cluster, NMBL, whose members were predominantly isolated from acute care center II. Our findings suggest that CR and dissemination of MBL clusters among P. aeruginosa populations in large geographic healthcare regions are dynamic processes that require continuous molecular surveillance. PMID- 17122003 TI - Identification of group A porcine rotavirus strains bearing a novel VP4 (P) Genotype in Italian swine herds. AB - The VP4 gene of a G5 Italian porcine rotavirus strain, 344/04-1, was nontypeable by PCR genotyping. The amino acid sequence of the full-length VP4 protein had low identity (or= 16 microg/ml), amikacin (MIC >or= 64 microg/ml), and ciprofloxacin (MIC >or= 4 microg/ml) were collected from 13 hospitals in the same prefecture in Japan. We also collected 70 clinical isolates of P. aeruginosa that were sensitive to one or more of these antibiotics and compared their characteristics with those of the MDR P. aeruginosa isolates. Of the 214 MDR P. aeruginosa isolates, 212 (99%) were serotype O11. We developed a loop-mediated isothermal amplification (LAMP) assay and a slide agglutination test for detection of the aac(6')-Iae gene and the AAC(6')-Iae protein, respectively. Of the 212 MDR P. aeruginosa isolates, 212 (100%) and 207 (98%) were positive in the LAMP assay and in the agglutination test, respectively. Mutations of gyrA and parC genes resulting in amino acid substitutions were detected in 213 of the 214 MDR P. aeruginosa isolates (99%). Of the 214 MDR P. aeruginosa isolates, 212 showed pulsed-field gel electrophoresis patterns with >or=70% similarity to that of IMCJ2.S1 and 83 showed a pattern identical to that of IMCJ2.S1, indicating that clonal expansion of MDR P. aeruginosa occurred in community hospitals in this area. The methods developed in this study to detect aac(6')-Iae were rapid and effective in diagnosing infections caused by various MDR P. aeruginosa clones. PMID- 17122010 TI - Evaluation of the sensitivities of DNA extraction and PCR methods for detection of Giardia duodenalis in stool specimens. AB - Sensitivities of DNA extraction methods and PCR methods for Giardia duodenalis were evaluated. A combination of the most sensitive methods, i.e., FTA filter paper and a PCR protocol using RH11/RH4 and GiarF/GiarR primers, showed no significant differences compared to immunofluorescence assay in terms of their sensitivities and specificities. PMID- 17122011 TI - Changing epidemiology of shigellosis and emergence of ciprofloxacin-resistant Shigellae in India. PMID- 17122012 TI - Urinary tract infection caused by Eikenella corrodens. PMID- 17122013 TI - Detection of human bocavirus in Canadian children in a 1-year study. AB - Human bocavirus was detected by PCR in 65 (5.1%) of 1,265 respiratory specimens collected in 2002 and 2003 from the Stollery Children's Hospital from children <17 years of age. The spectrum of illness included upper respiratory infection, croup, bronchiolitis, and pneumonia with a prominence of cough and fever. PMID- 17122014 TI - Antibiotic-resistant invasive pneumococcal clones in Italy. AB - A total of 105 multiple-antibiotic-resistant invasive pneumococcal isolates recovered in Italy from 2001 to 2003 were genetically characterized. Of these, 40 were penicillin-nonsusceptible (PNSSP) and 65 were penicillin-susceptible (PSSP) Streptococcus pneumoniae strains. Among the PNSSP isolates, 8 and 11 different restriction profiles were obtained for the pbp2b and pbp2x genes, respectively. Clonal groups were established on the basis of analysis of both pulsed-field gel electrophoresis (PFGE) types and multilocus sequence typing (MLST). Several international clones, such as Spain(23F)-1/ST81, Spain(6B)-2/ST90, Spain(9V) 3/ST156, and Sweden(15A)-25/ST63 [corrected] were identified among the PNSSP isolates. Other, smaller clones, such as the minor Spanish 19F clone/ST88 and Denmark(14)-32/ST230, were also found. Among the PSSP isolates, clones related to England(14)-9/ST9, Greece(6B)-22/ST273, and Portugal(19F)-21/ST177 were found. In addition, two large clones comprised nonvaccine serotypes. One, comprising serotype 3 isolates, corresponded to the clone Netherlands(3)-31/ST180; the other, comprising serotype 15B/C isolates, ST474, was not related to any previously described clone. Two small clusters related to the newly described clones Greece(21)-30/ST193 and Netherlands(15B)-37/ST199 included isolates with unrelated PFGE profiles. An unusual finding was the inability to obtain the MLST allelic profile for an isolate of serotype 19A, belonging to the Sweden(15A) 25/ST63 [corrected] clone, due to a large deletion of the xpt gene. Capsular switching was observed among both PNSSP and PSSP isolates and involved also serotypes not included in the 7-valent pneumococcal conjugate vaccine (PCV7), such as serotypes 15B/C and 19A. Since antibiotic-resistant nonvaccine serotype clones are present in Italy, continuous monitoring of pneumococcal epidemiology should be carried out in the PCV7 era. PMID- 17122015 TI - Globicatella sanguinis is an etiological agent of ventriculoperitoneal shunt associated meningitis. AB - Globicatella sanguinis is a very rare isolate in clinical samples. We present a case of meningitis in a 69-year-old female patient after implantation of an external left ventricular drainage due to a hydrocephalus. She recovered after antibiotic treatment with ceftriaxone. PMID- 17122016 TI - Controlled clinical comparison of VersaTREK and BacT/ALERT blood culture systems. AB - To assess the relative yields in automated microbial detection systems of bacteria and yeasts isolated from the blood of adult patients with suspected sepsis, we compared the new VersaTREK system (VTI) (TREK Diagnostic Systems, Cleveland, OH) to the BacT/ALERT 3D system (3D) (bioMerieux, Inc., Durham, NC). Identical protocols were followed for the two systems. Paired aerobic (REDOX 1) and anaerobic (REDOX 2) VTI media were compared with standard aerobic (SA) and anaerobic (SN) 3D media; each of the four culture bottles was filled with 6 to 9 ml of blood. All bottles flagged positive by the instruments were subcultured to determine both true-positive (growth) and false-positive (no growth) cultures. Additionally, to assess false-negative bottles, terminal subcultures were done on all negative companion bottles to true-positive bottles. All isolates were identified by standard methods. All 4 bottles were adequately filled and yielded 413 clinically significant isolates in 5,389 (79%) of the 6,786 4-bottle sets obtained. Although no overall difference in yield or in time to detection was detected between the two systems, significantly more streptococci and enterococci as a group were detected by VTI. Moreover, significantly more microorganisms were detected by VTI for patients receiving antimicrobial therapy. The two systems were comparable (P, not significant) at detecting the 179 unimicrobial episodes of bacteremia seen. False-positive rates for aerobic and anaerobic bottles, respectively, were 1.6% and 0.9% for VTI and 0.7% and 0.8% for 3D. We conclude that the VTI and 3D systems are comparable for detection of bloodstream infections with bacteria or yeasts. PMID- 17122017 TI - Isolation and antimicrobial susceptibilities of Chlamydial isolates from Western barred bandicoots. AB - A range of species of Chlamydiales have previously been detected in a variety of Australian marsupials, including koalas and western barred bandicoots. Thirty seven ocular, urogenital, or nasal swabs were obtained from 21 wild western barred bandicoots. Chlamydia culture and antibiotic susceptibility testing were performed for cycloheximide-treated HEp-2 cells in 96-well microtiter plates. Chlamydia spp. were isolated from 11 specimens from 9 (42.8%) bandicoots. All isolates were identified as Chlamydiales by conventional PCR with 16S and 23S rRNA gene primers specific to Chlamydiales and were confirmed to be Chlamydia pneumoniae by a C. pneumoniae-specific ompA-based real-time PCR assay and 16S rRNA and 23S rRNA gene signature sequence analyses. The MICs of azithromycin, doxycycline, ciprofloxacin, and enrofloxacin for 10 C. pneumoniae isolates from these bandicoots ranged from 0.015 to 1 microg/ml, 0.25 to 1 microg/ml, 0.25 to 2 microg/ml, and 0.25 to 0.5 microg/ml, respectively. The MICs at which 90% of isolates were inhibited and the minimal bactericidal concentrations were within the ranges reported previously for human isolates of C. pneumoniae. PMID- 17122018 TI - Mucor circinelloides was identified by molecular methods as a cause of primary cutaneous zygomycosis. AB - A case of primary cutaneous zygomycosis caused by Mucor circinelloides is described. Histopathology showed typical hyphae along with chlamydospores. The isolate was identified by molecular and phenotypic methods. The utility of sequence analysis of the internal transcribed spacer region is highlighted; however, further studies are needed to assess species genetic heterogeneity. PMID- 17122019 TI - Magnetic silica extraction for low-viremia human immunodeficiency virus type 1 genotyping. AB - Nucleic acid extraction and human immunodeficiency virus type 1 (HIV-1) genotyping using the NucliSens miniMAG platform and the TruGene HIV-1 genotyping kit gave HIV-1 sequence data from HIV-1-negative plasma spiked with 100 copies/ml reference HIV-1 RNA and from low-viremia clinical samples (<500 copies/ml) without the need for ultracentrifugation or nested second-round PCR. PMID- 17122020 TI - Usefulness of the BACTEC MGIT 960 system for isolation of mycobacterium tuberculosis from sputa subjected to long-term storage. AB - The recovery of Mycobacterium tuberculosis from sputa positive or negative for acid-fast bacilli that were stored for 17 +/- 7 days and inoculated in the BACTEC MGIT 960 system (MGIT) was higher than that from sputa inoculated in Lowenstein Jensen medium. MGIT is useful for isolation of M. tuberculosis from sputa subjected to long-term storage. PMID- 17122021 TI - Evidence for a link between parasite genotype and outcome of infection with Entamoeba histolytica. AB - The factors determining whether a person infected with Entamoeba histolytica develops disease remain obscure. To investigate whether the parasite genome contributes to the outcome, we have investigated the distribution of parasite genotypes among E. histolytica-infected individuals in Bangladesh. Samples were obtained from individuals who either were asymptomatic, had diarrhea/dysentery, or had developed a liver abscess. Genotypes were determined by using six tRNA linked polymorphic markers, and their distributions among the three sample groups were evaluated. A significant population differentiation in the genotype distribution was found for four of the six individual markers as well as for the combined genotypes, suggesting that the parasite genome does contribute in some way to the outcome of infection with E. histolytica. The markers themselves do not indicate the nature of the underlying genetic differences, but they may be linked to loci that do have an impact on the outcome of infection. PMID- 17122022 TI - Catabacter hongkongensis gen. nov., sp. nov., isolated from blood cultures of patients from Hong Kong and Canada. AB - Four bacterial isolates were recovered from the blood cultures of four patients, two of whom were from Hong Kong and two of whom were from Canada. The two Hong Kong strains were isolated from a 48-year-old man with intestinal obstruction and secondary sepsis (strain HKU16T) and from a 39-year-old man with acute appendicitis (strain HKU17), while the two Canadian strains were isolated from a 74-year-old man with biliary sepsis (strain CA1) and from a 66-year-old woman with metastatic carcinoma and sepsis (strain CA2). While the first three patients survived, the last patient died 2 weeks after the episode of bacteremia. All four isolates are strictly anaerobic, nonsporulating, gram-positive coccobacilli that were unidentified by conventional phenotypic tests and commercial identification systems. They grow on sheep blood agar as nonhemolytic pinpoint colonies after 48 h of incubation at 37 degrees C in an anaerobic environment. All are catalase positive and motile, with flagella. They produce acid from arabinose, glucose, mannose, and xylose. They do not produce indole or reduce nitrate. They are sensitive to penicillin, vancomycin, and metronidazole but resistant to cefotaxime. 16S rRNA gene sequence analysis showed 16.0%, 16.8%, and 21.0% base differences from Clostridium propionicum, Clostridium neopropionicum, and Atopobium minutum, respectively. The G+C content of strain HKU16T is 40.2% +/- 2.2%. Based on their phylogenetic affiliation, unique G+C content, and phenotypic characteristics, we propose a new genus and species, Catabacter hongkongensis gen. nov., sp. nov., to describe the bacterium, for which HKU16 is the type strain, and suggest that it be assigned to a new family, Catabacteriaceae. The gastrointestinal tract was probably the source of the bacterium for at least three of the four patients. The isolation of a catalase-positive, motile, nonsporulating, anaerobic gram-positive bacillus in clinical laboratories should raise the possibility of C. hongkongensis. Further studies should be performed to ascertain the epidemiology and other disease associations of this bacterium. PMID- 17122023 TI - Rapid detection of point mutations conferring resistance to fluoroquinolone in gyrA of Helicobacter pylori by allele-specific PCR. AB - Helicobacter pylori strains with reduced susceptibility to fluoroquinolones have a mutation at either codon 87 Asn or 91 Asp of the gyrA gene. A rapid test based on an allele-specific PCR (AS-PCR) was designed to detect the gyrA mutations. Clinical H. pylori isolates were obtained from the stomachs of 51 patients with H. pylori infections who showed treatment failure. The MICs of gatifloxacin (GAT) were determined by the agar dilution method. Identical genotyping results were obtained with AS-PCR and conventional PCR. The gyrA mutations of H. pylori causing reduced susceptibility to fluoroquinolones could be detected successfully by this method. A significant association was observed between the presence of mutations, as detected by AS-PCR, and the resistance of the strains to GAT. Moreover, genotyping by AS-PCR took less than 3 to 4 h. The AS-PCR method for the detection of gyrA mutations in H. pylori is useful for easy identification of fluoroquinolone-resistant strains of H. pylori. PMID- 17122024 TI - Prevalence and characterization of class I integrons among Pseudomonas aeruginosa and Acinetobacter baumannii isolates from patients in Nanjing, China. AB - Class I integrons were detected in 40.8% (40/98) of Pseudomonas aeruginosa strains and 52.8% (56/106) of Acinetobacter baumannii strains in the Nanjing area of China, including several cassette arrays not previously reported. PMID- 17122025 TI - Strain typing and determination of population structure of Candida krusei by multilocus sequence typing. AB - A multilocus sequence typing (MLST) scheme for Candida krusei was devised, based on sequencing of six gene fragments of the species. The existence of heterozygous results for each of the six fragments sequenced confirms that C. krusei is diploid for at least part of its genome. The C. krusei MLST scheme had a discriminatory index of 0.998, making this system ideal for strain typing of C. krusei clinical isolates. MLST data for 122 independent C. krusei isolates from a range of geographical sources were analyzed by eBURST, structure, and the unweighted-pair group method using average linkages to derive a population structure comprising four subtype strain clusters. There was no evidence of geographical associations with particular subtypes. Data for pairs of isolates from seven patients showed that each patient was colonized and/or infected with strain types that were indistinguishable by MLST. The C. krusei MLST database can be accessed online at http://pubmlst.org/ckrusei/. PMID- 17122026 TI - Nitric oxide and anandamide in OMT research. PMID- 17122027 TI - Unsubstantiated superiority claims for rivastigmine tartrate. PMID- 17122028 TI - Mechanisms of change: animal models in osteopathic medical research. PMID- 17122029 TI - Significance of differences in patency among cranial sutures. AB - OBJECTIVE: To evaluate the gross external characteristics of the coronal, lambdoid, and sagittal sutures in human cadaver skulls and determine if a difference exists in terms of patency, sex, and age. METHODS: The coronal, lambdoid, and sagittal sutures were described using a modified grading scale to quantify sutural patency. An open suture was graded as 0, a fused suture as 1, and an obliterated suture as 2, 3, or 4, depending on the extent of obliteration. RESULTS: Thirty-six skulls were examined, including 17 female and 19 male (age range, 56-101 y). When compared with the sagittal suture, the lambdoid suture was significantly more likely to be patent and least likely to be obliterated. No significant difference in suture grades was found between female and male skulls, and no significant difference was found between age and suture grade. CONCLUSION: The prolonged patency of the lambdoid suture may be due to external forces, such as the greater number of muscles affecting the lambdoid suture when compared with the sagittal suture. PMID- 17122030 TI - Incidence of iatrogenesis associated with osteopathic manipulative treatment of pediatric patients. AB - CONTEXT: Although studies have been published documenting the safety and efficacy of osteopathic manipulative treatment (OMT) in adults, no studies exist documenting the safety of OMT in pediatric patients. OBJECTIVE: To determine the incidence of iatrogenesis (ie, aggravations and complications) derived from OMT in the pediatric patient population. DESIGN: A retrospective review of medical records was conducted looking for documentation of aggravations or complications subsequent to OMT. Treatment-associated aggravations were defined as worsening of symptoms or complaints after treatment. Treatment complications were defined as cerebrovascular accidents, dislocation, fracture, pneumothorax, sprains and strains, or death as a treatment outcome. The authors documented all occurrences of treatment-associated aggravations or complications recorded at each office visit, as well as the timing of an aggravation between office visits. SETTING: Medical records of pediatric patients receiving OMT and being seen more than twice at osteopathic manipulative medicine offices in Pennsylvania and Virginia were reviewed. PATIENTS: Of 502 pediatric patients' medical records reviewed, 346 met the inclusion criteria of two or more office visits for which adequate follow up data were available. RESULTS: No treatment-associated complications were documented. Thirty-one (9%) patients had documented treatment-associated aggravations. There was no significant difference between the proportion of male and female patients considered in the study, relative to a 50/50 distribution (females, 48%; males, 52%). CONCLUSIONS: Osteopathic manipulative treatment appears to be a safe treatment modality in the pediatric population when administered by physicians with expertise in OMT. Future studies should be prospective and include larger numbers of patients to document the safety of OMT in this clinical application. PMID- 17122031 TI - Botulinum toxin in primary care medicine. AB - Clostridium botulinum, a gram-positive anaerobic bacterium, produces a potent neurotoxin that causes muscle paralysis. The therapeutic use of botulinum toxin was discovered in the 1970s and has since been used to treat patients with a broad range of medical complaints. Botulinum toxin (BTX) is used in the primary care setting to treat conditions such as allergic rhinitis, hyperhidrosis, lichen simplex chronicus, migraine, myofascial pain syndrome, and certain task-specific idiopathic focal dystonias (eg, writer's cramp)--in addition to its more publicized use for cosmetic enhancement of the face. The expanding range of therapeutic applications for BTX make it necessary for primary care physicians to understand the biochemistry, preparation, indications, and interactions of BTX. PMID- 17122032 TI - Patient confidentiality vs disclosure of inheritable risk: a survey-based study. AB - BACKGROUND: As the field of genetic medicine advances and more tests for genetic diseases become available, a dilemma of legal and ethical importance will be increasingly encountered by family physicians. Protecting the confidentiality of a patient with a genetic disease when the patient's family is at risk for inheriting the disease is a conflict that more and more physicians will be forced to address. METHODS: In March 2003, osteopathic family physicians in New Jersey were given a vignette in which a patient reveals that he has a genetic disease and demands that the information be kept confidential. The physicians were then given a 33-item questionnaire asking their opinions about disclosing an untreatable and a treatable disease to each of the patient's children and their mother, a former spouse. Also, physicians' opinions on larger issues were gathered, such as potential legal consequences of their actions and state laws in this area. Correlations between physicians' opinions and their demographic characteristics were also explored. Data were analyzed using Pearson product moment correlations and repeated-measures analyses of variance. RESULTS: Physicians tended to agree that adult children should be told if the disease were treatable, and these physicians were willing to accept responsibility for deciding whom to tell. With regard to the characters in the vignette, physicians felt comfortable telling the 22-year-old daughter, were unsure about telling the 17-year-old son, and would not tell either a former spouse or the 12-year-old son. The respondents agreed that state laws should permit disclosure rather than require it, and they did not think state laws should forbid it. CONCLUSION: The authors recommend that states draft a definitive public policy about when, how, and whether to disclose pertinent medical information to those at risk for inheriting a genetic disease. PMID- 17122033 TI - Asymptomatic thrombus trapping in vena cava filters: a case series. AB - The prophylactic use of vena cava filters (VCFs) remains controversial. Class I data supporting the placement of prophylactic VCFs are lacking. In addition, there are concerns regarding the long-term effects of permanent VCF placement in patients with a relatively brief period of thromboembolic risk. Venous thromboembolism can be a lethal event, occurring without a prodrome. Yet, this condition is potentially preventable. The efficacy of VCFs is difficult to determine, as it is likely that when a VCF actually traps a thrombus, the event is clinically asymptomatic. We present three cases of asymptomatic thrombus trapping in VCFs, recognized through incidental radiographic studies. PMID- 17122034 TI - Uniform serial sectioning for transmission electron microscopy. PMID- 17122035 TI - Reversible disruption of dynactin 1-mediated retrograde axonal transport in polyglutamine-induced motor neuron degeneration. AB - Spinal and bulbar muscular atrophy (SBMA) is a hereditary neurodegenerative disease caused by an expansion of a trinucleotide CAG repeat encoding the polyglutamine tract in the androgen receptor (AR) gene. To elucidate the pathogenesis of polyglutamine-mediated motor neuron dysfunction, we investigated histopathological and biological alterations in a transgenic mouse model of SBMA carrying human pathogenic AR. In affected mice, neurofilaments and synaptophysin accumulated at the distal motor axon. A similar intramuscular accumulation of neurofilament was detected in the skeletal muscle of SBMA patients. Fluoro-gold labeling and sciatic nerve ligation demonstrated an impaired retrograde axonal transport in the transgenic mice. The mRNA level of dynactin 1, an axon motor for retrograde transport, was significantly reduced in the SBMA mice resulting from pathogenic AR-induced transcriptional dysregulation. These pathological events were observed before the onset of neurological symptoms, but were reversed by castration, which prevents nuclear accumulation of pathogenic AR. Overexpression of dynactin 1 mitigated neuronal toxicity of the pathogenic AR in a cell culture model of SBMA. These observations indicate that polyglutamine-dependent transcriptional dysregulation of dynactin 1 plays a crucial role in the reversible neuronal dysfunction in the early stage of SBMA. PMID- 17122036 TI - Octopamine mediates thermal preconditioning of the locust ventilatory central pattern generator via a cAMP/protein kinase A signaling pathway. AB - We investigated the role of biogenic amines in generating thermoprotection of the ventilatory motor pattern circuitry in Locusta migratoria. Levels of octopamine (OA) and dopamine (DA) in the metathoracic ganglion decreased during heat stress. We measured the thermosensitivity of central pattern generation in response to a ramped increase of temperature in semi-intact preparations. OA, DA, and tyramine (TA) were either bath applied or injected into the locust hemocoel 4-8 h before testing. Neither TA nor DA modified the thermotolerance of ventilatory motor pattern generation. However, OA treatment by bath applications (10(-4) M OA) or by injections into the hemocoel (2 microg/10 microl OA) mimicked heat shock preconditioning and improved the thermotolerance of the motor pattern by increasing the failure temperature and by decreasing the time taken to recover operation after a return to room temperature. Heat shock-induced thermoprotection was eradicated in locusts preinjected with epinastine (Oct betaR antagonist). Neuropil injections of the cAMP agonist and protein kinase A (PKA) activator, Sp cAMPs, both conferred thermoprotection in control locusts and rescued thermoprotection in epinastine-treated HS locusts. Similar injections of the PKA inhibitor Rp-cAMPs blocked the thermoprotective effect of bath-applied OA. Octopamine-mediated thermoprotection was also abolished with neuropil injections of cycloheximide or actinomycin D, indicating a requirement for transcription and translation. We conclude that OA has a crucial role in triggering protein synthesis-dependent physiological adaptations to protect CNS function during heat stress by activating a cAMP/PKA pathway. PMID- 17122037 TI - Regional interaction of endoplasmic reticulum Ca2+ signals between soma and dendrites through rapid luminal Ca2+ diffusion. AB - The endoplasmic reticulum (ER) Ca2+ store plays a key role in integration and conveyance of Ca2+ signals in highly polarized neurons. The interconnected ER network in neurons generates Ca2+ signals in local domains, but the regional interaction is unclear. Here, we show that continuous or repetitive applications of caffeine produced robust Ca2+ release from the ER Ca2+ store in dendritic areas without severe store depletion, but that similar stimuli applied to soma caused rapid store depletion in acutely isolated midbrain dopamine neurons. Partial emptying of the ER Ca2+ store within a dendrite caused a similar level of store depletion in unstimulated dendrites, as well as in soma. Photobleaching and local stimulation experiments revealed that Ca2+ and the dye trapped within the ER diffused rapidly from the soma to dendrites up to 90 microm, which we could resolve, suggesting that the ER network acts as a functional tunnel for rapid Ca2+ transport. These data imply that the ER in soma acts as a Ca2+ reservoir supplying Ca2+ to the dendritic store, and that the dendritic store, hence, is able to respond to Ca2+-mobilizing input signals endurably. PMID- 17122038 TI - Unanticipated region- and cell-specific downregulation of individual KChIP auxiliary subunit isotypes in Kv4.2 knock-out mouse brain. AB - Kv4 family voltage-gated potassium channel alpha subunits and Kv channel interacting protein (KChIP) and dipeptidyl aminopeptidase-like protein subunits comprise somatodendritic A-type channels in mammalian neurons. Recently, a mouse was generated with a targeted deletion of Kv4.2, a Kv4 alpha subunit expressed in many but not all mammalian brain neurons. Kv4.2-/- mice are grossly indistinguishable from wild-type (WT) littermates. Here we used immunohistochemistry to analyze expression of component Kv4 and KChIP subunits of A-type channels in WT and Kv4.2-/- brains. We found that the expression level, and cellular and subcellular distribution of the other prominent brain Kv4 family member Kv4.3, was indistinguishable between WT and Kv4.2-/- samples. However, we found unanticipated regional and cell-specific decreases in expression of KChIPs. The degree of altered expression of individual KChIP isoforms in different regions and neurons precisely follows the level of Kv4.2 normally found at those sites and presumably their extent of association of these KChIPs with Kv4.2. The dramatic effects of Kv4.2 deletion on KChIP expression suggest that, in addition to previously characterized effects of KChIPs on the functional properties, trafficking, and turnover rate of Kv4 channels, Kv4:KChIP association may confer reciprocal Kv4.2-dependent effects on KChIPs. The impact of Kv4.2 deletion on KChIP expression also supports the major role of KChIPs as auxiliary subunits of Kv4 channels. PMID- 17122039 TI - Deletion of Kv4.2 gene eliminates dendritic A-type K+ current and enhances induction of long-term potentiation in hippocampal CA1 pyramidal neurons. AB - Dendritic, backpropagating action potentials (bAPs) facilitate the induction of Hebbian long-term potentiation (LTP). Although bAPs in distal dendrites of hippocampal CA1 pyramidal neurons are attenuated when propagating from the soma, their amplitude can be increased greatly via downregulation of dendritic A-type K+ currents. The channels that underlie these currents thus may represent a key regulatory component of the signaling pathways that lead to synaptic plasticity. We directly tested this hypothesis by using Kv4.2 knock-out mice. Deletion of the Kv4.2 gene and a loss of Kv4.2 protein resulted in a specific and near-complete elimination of A-type K+ currents from the apical dendrites of CA1 pyramidal neurons. The absence of dendritic Kv4.2-encoded A-type K+ currents led to an increase of bAP amplitude and an increase of concurrent Ca2+ influx. Furthermore, CA1 pyramidal neurons lacking dendritic A-type K+ currents from Kv4.2 knock-out mice exhibited a lower threshold than those of wild-type littermates for LTP induction with the use of a theta burst pairing protocol. LTP triggered with the use of a saturating protocol, on the other hand, remained indistinguishable between Kv4.2 knock-out and wild-type neurons. Our results support the hypothesis that dendritic A-type K+ channels, composed of Kv4.2 subunits, regulate action potential backpropagation and the induction of specific forms of synaptic plasticity. PMID- 17122040 TI - Intracellular and trans-synaptic regulation of glutamatergic synaptogenesis by EphB receptors. AB - The majority of mature excitatory synapses in the CNS are found on dendritic spines and contain AMPA- and NMDA-type glutamate receptors apposed to presynaptic specializations. EphB receptor tyrosine kinase signaling has been implicated in both NMDA-type glutamate receptor clustering and dendritic spine formation, but it remains unclear whether EphB plays a broader role in presynaptic and postsynaptic development. Here, we find that EphB2 is involved in organizing excitatory synapses through the independent activities of particular EphB2 protein domains. We demonstrate that EphB2 controls AMPA-type glutamate receptor localization through PDZ (postsynaptic density-95/Discs large/zona occludens-1) binding domain interactions and triggers presynaptic differentiation via its ephrin binding domain. Knockdown of EphB2 in dissociated neurons results in decreased functional synaptic inputs, spines, and presynaptic specializations. Mice lacking EphB1-EphB3 have reduced numbers of synapses, and defects are rescued with postnatal reexpression of EphB2 in single neurons in brain slice. These results demonstrate that EphB2 acts to control the organization of specific classes of mature glutamatergic synapses. PMID- 17122041 TI - Chronic pain and the emotional brain: specific brain activity associated with spontaneous fluctuations of intensity of chronic back pain. AB - Living with unrelenting pain (chronic pain) is maladaptive and is thought to be associated with physiological and psychological modifications, yet there is a lack of knowledge regarding brain elements involved in such conditions. Here, we identify brain regions involved in spontaneous pain of chronic back pain (CBP) in two separate groups of patients (n = 13 and n = 11), and contrast brain activity between spontaneous pain and thermal pain (CBP and healthy subjects, n = 11 each). Continuous ratings of fluctuations of spontaneous pain during functional magnetic resonance imaging were separated into two components: high sustained pain and increasing pain. Sustained high pain of CBP resulted in increased activity in the medial prefrontal cortex (mPFC; including rostral anterior cingulate). This mPFC activity was strongly related to intensity of CBP, and the region is known to be involved in negative emotions, response conflict, and detection of unfavorable outcomes, especially in relation to the self. In contrast, the increasing phase of CBP transiently activated brain regions commonly observed for acute pain, best exemplified by the insula, which tightly reflected duration of CBP. When spontaneous pain of CBP was contrasted to thermal stimulation, we observe a double-dissociation between mPFC and insula with the former correlating only to intensity of spontaneous pain and the latter correlating only to pain intensity for thermal stimulation. These findings suggest that subjective spontaneous pain of CBP involves specific spatiotemporal neuronal mechanisms, distinct from those observed for acute experimental pain, implicating a salient role for emotional brain concerning the self. PMID- 17122042 TI - Interleukin-11 potentiates oligodendrocyte survival and maturation, and myelin formation. AB - Mechanisms that regulate oligodendrocyte survival and myelin formation are an intense focus of research into myelin repair in the lesions of multiple sclerosis (MS). Although demyelination and oligodendrocyte loss are pathological hallmarks of the disease, increased oligodendrocyte numbers and remyelination are frequently observed in early lesions, but these diminish as the disease course progresses. In the current study, we used a microarray-based approach to investigate genes regulating repair in MS lesions, and identified interleukin-11 (IL-11) as an astrocyte-derived factor that potentiates oligodendrocyte survival and maturation, and myelin formation. IL-11 was induced in human astrocyte cultures by the cytokines IL-1beta and TGFbeta1, which are both prominently expressed in MS plaques. In MS tissue samples, IL-11 was expressed by reactive astrocytes, with expression particularly localized at the myelinated border of both active and silent lesions. Its receptor, IL-11R alpha, was expressed by oligodendrocytes. In experiments in human cultures in vitro, IL-11R alpha localized to immature oligodendrocytes, and its expression decreased during maturation. In cultures treated with IL-11, we observed a significant increase in oligodendrocyte number, and this was associated with enhanced oligodendrocyte survival and maturation. Importantly, we also found that IL-11 treatment was associated with significantly increased myelin formation in rodent CNS cocultures. These data are the first to implicate IL-11 in oligodendrocyte viability, maturation, and myelination. We suggest that this pathway may represent a potential therapeutic target for oligodendrocyte protection and remyelination in MS. PMID- 17122043 TI - Why does brain damage impair memory? A connectionist model of object recognition memory in perirhinal cortex. AB - Object recognition is the canonical test of declarative memory, the type of memory putatively impaired after damage to the temporal lobes. Studies of object recognition memory have helped elucidate the anatomical structures involved in declarative memory, indicating a critical role for perirhinal cortex. We offer a mechanistic account of the effects of perirhinal cortex damage on object recognition memory, based on the assumption that perirhinal cortex stores representations of the conjunctions of visual features possessed by complex objects. Such representations are proposed to play an important role in memory when it is difficult to solve a task using representations of only individual visual features of stimuli, thought to be stored in regions of the ventral visual stream caudal to perirhinal cortex. The account is instantiated in a connectionist model, in which development of object representations with visual experience provides a mechanism for judgment of previous occurrence. We present simulations addressing the following empirical findings: (1) that impairments after damage to perirhinal cortex (modeled by removing the "perirhinal cortex" layer of the network) are exacerbated by lengthening the delay between presentation of to-be-remembered items and test, (2) that such impairments are also exacerbated by lengthening the list of to-be-remembered items, and (3) that impairments are revealed only when stimuli are trial unique rather than repeatedly presented. This study shows that it may be possible to account for object recognition impairments after damage to perirhinal cortex within a hierarchical, representational framework, in which complex conjunctive representations in perirhinal cortex play a critical role. PMID- 17122044 TI - Stimulus frequency processing in awake rat barrel cortex. AB - In awake rats, we examined the relationship between neural spiking activity in primary somatic sensory cortex and the frequency of whisker stimulation. Neural responses were recorded extracellularly in barrel cortex while single whiskers were deflected with 0.5-18 air puffs per second (apps), a range that includes the whisk rates observed when rats explore their environment and discriminate surfaces with their whiskers. Twenty-nine neurons in layers III and IV were isolated in three rats (23 in barrel columns and 6 in septum columns). At < or = 9 apps, cortical neurons responded with one to two spikes per stimulus, whereas at > 9 apps, the response efficacy was reduced to only 0.2-0.4 spikes per stimulus. Several mechanisms are discussed that could account for the decrement in responsiveness. Despite this adaptation, neural spike rates increased in direct proportion with stimulus frequency when cast on logarithmic scales. At > 9 apps, however, this relationship deteriorated in barrel columns in which the response approximately halved. In contrast, septum column cells continued to increase their spike rates linearly up to 18 apps, although they responded at lower magnitude than the barrel column cells. Our findings suggest that septum column neurons are potential candidates to encode stimulus frequency using spike rate across the entire frequency range relevant to rats' whisking behavior. PMID- 17122045 TI - Non-Gaussian membrane potential dynamics imply sparse, synchronous activity in auditory cortex. AB - Many models of cortical dynamics have focused on the high-firing regime, in which neurons are driven near their maximal rate. Here we consider the responses of neurons in auditory cortex under typical low-firing rate conditions, when stimuli have not been optimized to drive neurons maximally. We used whole-cell patch clamp recording in vivo to measure subthreshold membrane potential fluctuations in rat primary auditory cortex in both the anesthetized and awake preparations. By analyzing the subthreshold membrane potential dynamics on single trials, we made inferences about the underlying population activity. We found that, during both spontaneous and evoked responses, membrane potential was highly non Gaussian, with dynamics consisting of occasional large excursions (sometimes tens of millivolts), much larger than the small fluctuations predicted by most random walk models that predict a Gaussian distribution of membrane potential. Thus, presynaptic inputs under these conditions are organized into quiescent periods punctuated by brief highly synchronous volleys, or "bumps." These bumps were typically so brief that they could not be well characterized as "up states" or "down states." We estimate that hundreds, perhaps thousands, of presynaptic neurons participate in the largest volleys. These dynamics suggest a computational scheme in which spike timing is controlled by concerted firing among input neurons rather than by small fluctuations in a sea of background activity. PMID- 17122046 TI - A circadian clock in the olfactory bulb controls olfactory responsivity. AB - Recently, it has been shown that multiple mammalian cell types express daily rhythms in vitro. Although the suprachiasmatic nucleus (SCN) of the hypothalamus is known to regulate a wide range of circadian behaviors, the role for intrinsic rhythmicity in other tissues is unknown. We tested whether the main olfactory bulb (OB) of mice mediates daily changes in olfaction. We found circadian rhythms in cedar oil-induced c-Fos, a protein marker of cellular excitation, in the mitral and granular layers of the OB and in the piriform cortex (PC). These oscillations persisted in constant darkness with a fourfold change in amplitude and a peak approximately 4 h after the onset of daily locomotor activity. Electrolytic lesions of the SCN abolished circadian locomotor rhythms, but not odor-induced c-Fos rhythms in the OB or PC. Furthermore, removal of the OB abolished spontaneous circadian cycling of c-Fos in the PC, shortened the free running period of locomotor rhythms, and accelerated re-entrainment after a 6 h advance and slowed re-entrainment after a 6 h delay in the light schedule. OB ablation or odorant altered the amplitude of c-Fos rhythms in the SCN and ablation of one OB abolished c-Fos rhythms in the ipsilateral PC, but not in the contralateral OB and PC. We conclude that the OB comprises a master circadian pacemaker, which enhances olfactory responsivity each night, drives rhythms in the PC, and interacts with the SCN to coordinate other daily behaviors. PMID- 17122047 TI - Molecular markers of neuronal progenitors in the embryonic cerebellar anlage. AB - The cerebellum, like the cerebrum, includes a nuclear structure and an overlying cortical structure. Experiments in the past decade have expanded knowledge beyond the traditional function of the cerebellum to include critical roles in motor learning and memory and sensory discrimination. The initial steps in cerebellar development depend on inductive signaling involving FGF and Wnt proteins produced at the mesencephalic/metencephalic boundary. To address the issue of how individual cerebellar cell fates within the cerebellar territory are specified, we examined the expression of transcription factors, including mammalian homologues of LIM homeodomain-containing proteins, basic helix-loop-helix proteins, and three amino acid loop-containing proteins. The results of these studies show that combinatorial codes of transcription factors define precursors of the cerebellar nuclei, and both Purkinje cells and granule neurons of the cerebellar cortex. Examination of gene expression patterns in several hundred lines of Egfp-BAC (bacterial artificial chromosome) transgenic mice in the GENSAT Project revealed numerous genes with restricted expression in cerebellar progenitor populations, including genes specific for cerebellar nuclear precursors and Purkinje cell precursors. In addition, we identified patterns of gene expression that link granule and Purkinje cells to their precerebellar nuclei. These results identify molecular pathways that offer new insights on the development of the nuclear and cortical structures of the cerebellum, as well as components of the cerebellar circuitry. PMID- 17122048 TI - Integration of new neurons into functional neural networks. AB - Although it is established that new granule cells can be born and can survive in the adult mammalian hippocampus, there remains some question concerning the functional integration of these neurons into behaviorally relevant neural networks. By using high-resolution confocal microscopy, we have applied a new strategy to address the question of functional integration of newborn neurons into networks that mediate spatial information processing and memory formation. Exploration-induced expression of the immediate-early gene Arc in hippocampal cells has been linked to cellular activity observed in electrophysiological recordings under the same behavioral conditions. We investigated whether mature (5-month-old), newborn granule cells express Arc in response to a discrete spatial experience by detecting the expression of Arc in combination with NeuN (neuron-specific nuclear protein)-positive and bromodeoxyuridine-positive cells. We found that mature new granule cells do indeed express Arc in response to an exploration experience, supporting the idea that these cells are well integrated into hippocampal circuits. The proportion of mature newborn neurons that expressed Arc in response to exploration, however, was significantly higher (approximately 2.8%) than the proportion of cells that expressed Arc in the already existing population of granule cells (approximately 1.6%; p < 0.01). This finding extends previous data suggesting that the cellular physiology of newborn granule neurons differs from that of the existing population by indicating that these properties are retained in mature adult-generated neurons. Thus, these data have interesting implications for network models of spatial information processing and the role of hippocampal circuits in memory, indicating that mature new neurons are selectively recruited into hippocampal cell assemblies in higher proportions than older cells. PMID- 17122049 TI - The Nogo-Nogo receptor pathway limits a spectrum of adult CNS axonal growth. AB - The hypothesis that Nogo-A (Reticulon 4A) and Nogo-66 receptor (NgR1) limit adult CNS axonal growth after injury is supported by both in vitro experiments and in vivo pharmacological studies. However, genetic assessment of the role of Nogo-A in corticospinal tract (CST) axons after spinal cord dorsal hemisection has yielded conflicting results. CST regeneration is detected in homozygous nogo ab(trap/trap) mice, but not in nogo-ab(atg/atg) mice. CST regeneration is also present after pharmacological NgR blockade, but not in ngr1(-/-) mice. To assess the nogo-ab(atg) and ngr1-null alleles for other axon growth phenotypes, we created unilateral pyramidotomies and monitored the uninjured CST. There is robust pyramidotomy-induced growth of nogo-ab(atg/atg) and ngr1(-/-) CST axons into denervated cervical gray matter. This fiber growth correlates with recovery of fine motor skill in the affected forelimb. Thus nogo-ab and ngr1 play a modulated role in limiting CNS axonal growth across a spectrum of different tracts in various lesion models. PMID- 17122050 TI - Distinct populations of spinal cord lamina II interneurons expressing G-protein gated potassium channels. AB - Noxious stimuli are sensed and carried to the spinal cord dorsal horn by A delta and C primary afferent fibers. Some of this input is relayed directly to supraspinal sites by projection neurons, whereas much of the input impinges on a heterogeneous population of interneurons in lamina II. Previously, we demonstrated that G-protein-gated inwardly rectifying potassium (GIRK) channels are expressed in lamina II of the mouse spinal cord and that pharmacologic ablation of spinal GIRK channels selectively blunts the analgesic effect of high but not lower doses of intrathecal mu-opioid receptor (MOR) agonists. Here, we report that GIRK channels formed by GIRK1 and GIRK2 subunits are found in two large populations of lamina II excitatory interneurons. One population displays relatively large apparent whole-cell capacitances and prominent GIRK-dependent current responses to the MOR agonist [D-Ala2,N-MePhe4,Gly-ol5] -enkephalin (DAMGO). A second population shows smaller apparent capacitance values and a GIRK dependent response to the GABA(B) receptor agonist baclofen, but not DAMGO. Ultrastructural analysis revealed that GIRK subunits preferentially label type I synaptic glomeruli, suggesting that GIRK-containing lamina II interneurons receive prominent input from C fibers, while receiving little input from A delta fibers. Thus, excitatory interneurons in lamina II of the mouse spinal cord can be subdivided into different populations based on the neurotransmitter system coupled to GIRK channels. This important distinction will afford a unique opportunity to characterize spinal nociceptive circuitry with defined physiological significance. PMID- 17122051 TI - Cross-modal processing in early visual and auditory cortices depends on expected statistical relationship of multisensory information. AB - Previous studies have shown that processing information in one sensory modality can either be enhanced or attenuated by concurrent stimulation of another modality. Here, we reconcile these apparently contradictory results by showing that the sign of cross-modal interactions depends on whether the content of two modalities is associated or not. When concurrently presented auditory and visual stimuli are paired by chance, cue-induced preparatory neural activity is strongly enhanced in the task-relevant sensory system and suppressed in the irrelevant system. Conversely, when information in the two modalities is reliably associated, activity is enhanced in both systems regardless of which modality is task relevant. Our findings illustrate an ecologically optimal flexibility of the neural mechanisms that govern multisensory processing: facilitation occurs when integration is expected, and suppression occurs when distraction is expected. Because thalamic structures were more active when the senses needed to operate separately, we propose them to serve gatekeeper functions in early cross-modal interactions. PMID- 17122052 TI - The substantia nigra pars compacta and temporal processing. AB - The basal ganglia and cerebellum are considered to play a role in timing, although their differential roles in timing remain unclear. It has been proposed that the timing of short milliseconds-range intervals involves the cerebellum, whereas longer seconds-range intervals engage the basal ganglia (Ivry, 1996). We tested this hypothesis using positron emission tomography to measure regional cerebral blood flow in eight right-handed males during estimation and reproduction of long and short intervals. Subjects performed three tasks: (1) reproduction of a short 500 ms interval, (2) reproduction of a long 2 s interval, and (3) a control simple reaction time (RT) task. We compared the two time reproduction tasks with the control RT task to investigate activity associated with temporal processing once additional cognitive, motor, or sensory processing was controlled. We found foci in the left substantia nigra and the left lateral premotor cortex to be significantly more activated in the time reproduction tasks than the control RT task. The left caudate nucleus and right cerebellum were more active in the short relative to the long interval, whereas greater activation of the right putamen and right cerebellum occurred in the long rather than the short interval. These results suggest that the basal ganglia and the cerebellum are engaged by reproduction of both long and short intervals but play different roles. The fundamental role of the substantia nigra in temporal processing is discussed in relation to previous animal lesion studies and evidence for the modulating influence of dopamine on temporal processing. PMID- 17122053 TI - Differential expression of I(A) channel subunits Kv4.2 and Kv4.3 in mouse visual cortical neurons and synapses. AB - In cortical neurons, pore-forming alpha-subunits of the Kv4 subfamily underlie the fast transient outward K+ current (I(A)). Considerable evidence has accumulated demonstrating specific roles for I(A) channels in the generation of individual action potentials and in the regulation of repetitive firing. Although I(A) channels are thought to play a role in synaptic processing, little is known about the cell type- and synapse-specific distribution of these channels in cortical circuits. Here, we used immunolabeling with specific antibodies against Kv4.2 and Kv4.3, in combination with GABA immunogold staining, to determine the cellular, subcellular, and synaptic localization of Kv4 channels in the primary visual cortex of mice, in which subsets of pyramidal cells express yellow fluorescent protein. The results show that both Kv4.2 and Kv4.3 are concentrated in layer 1, the bottom of layer 2/3, and in layers 4 and 5/6. In all layers, clusters of Kv4.2 and Kv4.3 immunoreactivity are evident in the membranes of the somata, dendrites, and spines of pyramidal cells and GABAergic interneurons. Electron microscopic analyses revealed that Kv4.2 and Kv4.3 clusters in pyramidal cells and interneurons are excluded from putative excitatory synapses, whereas postsynaptic membranes at GABAergic synapses often contain Kv4.2 and Kv4.3. The presence of Kv4 channels at GABAergic synapses would be expected to weaken inhibition during dendritic depolarization by backpropagating action potentials. The extrasynaptic localization of Kv4 channels near excitatory synapses, in contrast, should stabilize synaptic excitation during dendritic depolarization. Thus, the synapse-specific distribution of Kv4 channels functions to optimize dendritic excitation and the association between presynaptic and postsynaptic activity. PMID- 17122054 TI - Ca2+ dynamics along identified synaptic terminals in Drosophila larvae. AB - Changes in intracellular Ca2+ concentration ([Ca2+]i) play an important role in the function and plasticity of synapses. We characterized the changes in [Ca2+]i produced by action potentials (APs) along two identified motor terminals found on separate muscle fibers in Drosophila larvae and examined factors that influence the amplitude and duration of the residual Ca2+ signal. We were able to measure Ca2+ transients produced along terminals by both single APs and AP trains using Oregon Green 488 BAPTA-1 and streaming images at 20-50 Hz. The decay of [Ca2+]i after single APs or AP trains was well fit by a single exponential. For single APs, the Ca2+ transient amplitude and decay rate were similar at boutons and bottleneck regions and much smaller at the axon. Also, the amplitude of single-AP Ca2+ transients was inversely correlated with bouton width. During AP trains, the increase in [Ca2+]i became more uniform: the difference in boutons and axons was reduced, and the increase in [Ca2+]i was not correlated with bouton width. The [Ca2+]i decay tau was directly correlated with bouton width for both single APs and AP trains. For one terminal, distal boutons had larger single-AP Ca2+ transients than proximal ones, probably attributable to greater Ca2+ influx for distal boutons. Pharmacological studies showed that Ca2+ clearance from these synaptic terminals after single APs and AP trains was primarily attributable to Ca2+ extrusion by the plasma membrane Ca2+ ATPase (PMCA). Immunostaining of larval muscle fibers showed high levels of the PMCA at the neuromuscular junction. PMID- 17122055 TI - Vesicular glutamate transporter 2 is required for central respiratory rhythm generation but not for locomotor central pattern generation. AB - Glutamatergic excitatory neurotransmission is dependent on glutamate release from presynaptic vesicles loaded by three members of the solute carrier family, Slc17a6-8, which function as vesicular glutamate transporters (VGLUTs). Here, we show that VGLUT2 (Slc17a6) is required for life ex utero. Vglut2 null mutant mice die immediately after birth because of the absence of respiratory behavior. Investigations at embryonic stages revealed that neural circuits in the location of the pre-Botzinger (PBC) inspiratory rhythm generator failed to become active. However, neurons with bursting pacemaker properties and anatomical integrity of the PBC area were preserved. Vesicles at asymmetric synapses were fewer and malformed in the Vglut2 null mutant hindbrain, probably causing the complete disruption of AMPA/kainate receptor-mediated synaptic activity in mutant PBC cells. The functional deficit results from an inability of PBC neurons to achieve synchronous activation. In contrast to respiratory rhythm generation, the locomotor central pattern generator of Vglut2 null mutant mice displayed normal rhythmic and coordinated activity, suggesting differences in their operating principles. Hence, the present study identifies VGLUT2-mediated signaling as an obligatory component of the developing respiratory rhythm generator. PMID- 17122056 TI - Long-term neuroadaptations produced by withdrawal from repeated cocaine treatment: role of dopaminergic receptors in modulating cortical excitability. AB - Dopamine (DA) modulates neuronal activity in the prefrontal cortex (PFC) and is necessary for optimal cognitive function. Dopamine transmission in the PFC is also important for the behavioral adaptations produced by repeated exposure to cocaine. Therefore, we investigated the effects of repeated cocaine treatment followed by withdrawal (2-4 weeks) on the responsivity of cortical cells to electrical stimulation of the ventral tegmental area (VTA) and to systemic administration of DA D1 or D2 receptor antagonists. Cortical cells in cocaine- and saline-treated animals exhibited a similar decrease in excitability after the administration of D1 receptor antagonists. In contrast, cortical neurons from cocaine-treated rats exhibited a lack of D2-mediated regulation relative to saline rats. Furthermore, in contrast to saline-treated animals, VTA stimulation did not increase cortical excitability in the cocaine group. These data suggest that withdrawal from repeated cocaine administration elicits some long-term neuroadaptations in the PFC, including (1) reduced D2-mediated regulation of cortical excitability, (2) reduced responsivity of cortical cells to phasic increases in DA, and (3) a trend toward an overall decrease in excitability of PFC neurons. PMID- 17122057 TI - Abnormal Ca2+ dynamics in transgenic mice with neuron-specific mitochondrial DNA defects. AB - Maintenance of mitochondrial DNA (mtDNA) depends on nuclear-encoded proteins such as mtDNA polymerase (POLG), whose mutations are involved in the diseases caused by mtDNA defects including mutation and deletion. The defects in mtDNA and in intracellular Ca2+ ([Ca2+]i) homeostasis have been reported in bipolar disorder (BD). To understand the relevance of the mtDNA defects to BD, we studied transgenic (Tg) mice in which mutant POLG (mutPOLG) was expressed specifically in neurons. mtDNA defects were accumulated in the brains of mutPOLG Tg mice in an age-dependent manner and the mutant mice showed BD-like behavior. However, the molecular and cellular basis for the abnormalities has not been clarified. In this study, we investigated Ca2+ regulation by isolated mitochondria and [Ca2+]i dynamics in the neurons of mutPOLG Tg mice. Mitochondria from the mutant mice sequestered Ca2+ more rapidly, whereas Ca2+ retention capacity and membrane potential, a driving force of Ca2+ uptake, of mitochondria were unaffected. To elucidate the molecular mechanism of the altered Ca2+ uptake, we performed DNA microarray analysis and found that the expression of cyclophilin D (CyP-D), a component of the permeability transition pore, was downregulated in the brains of mutPOLG Tg mice. Cyclosporin A, an inhibitor of CyP-D, mimicked the enhanced Ca2+ uptake in mutant mice. Furthermore, G-protein-coupled receptor-mediated [Ca2+]i increase was attenuated in hippocampal neurons of the mutant mice. These findings suggest that mtDNA defects lead to enhancement of Ca2+ uptake rate via CyP-D downregulation and alter [Ca2+]i dynamics, which may be involved in the pathogenesis of BD. PMID- 17122058 TI - Somatodendritic Kv7/KCNQ/M channels control interspike interval in hippocampal interneurons. AB - The M-current (I(M)), comprised of Kv7 channels, is a voltage-activated K+ conductance that plays a key role in the control of cell excitability. In hippocampal principal cells, I(M) controls action potential (AP) accommodation and contributes to the medium-duration afterhyperpolarization, but the role of I(M) in control of interneuron excitability remains unclear. Here, we investigated I(M) in hippocampal stratum oriens (SO) interneurons, both from wild type and transgenic mice in which green fluorescent protein (GFP) was expressed in somatostatin-containing interneurons. Somatodendritic expression of Kv7.2 or Kv7.3 subunits was colocalized in a subset of GFP+ SO interneurons, corresponding to oriens-lacunosum moleculare (O-LM) cells. Under voltage clamp (VC) conditions at -30 mV, the Kv7 channel antagonists linopirdine/XE-991 abolished the I(M) amplitude present during relaxation from -30 to -50 mV and reduced the holding current (I(hold)). In addition, 0.5 mM tetraethylammonium reduced I(M), suggesting that I(M) was composed of Kv7.2-containing channels. In contrast, the Kv7 channel opener retigabine increased I(M) amplitude and I(hold). When strongly depolarized in VC, the linopirdine-sensitive outward current activated rapidly and comprised up to 20% of the total current. In current-clamp recordings from GFP+ SO cells, linopirdine induced depolarization and increased AP frequency, whereas retigabine induced hyperpolarization and arrested firing. In multicompartment O-LM interneuron models that incorporated I(M), somatodendritic placement of Kv7 channels best reproduced experimentally measured I(M). The models suggest that Kv3- and Kv7-mediated channels both rapidly activate during single APs; however, Kv3 channels control rapid repolarization of the AP, whereas Kv7 channels primarily control the interspike interval. PMID- 17122059 TI - Mice with conditional inactivation of fibroblast growth factor receptor-2 signaling in oligodendrocytes have normal myelin but display dramatic hyperactivity when combined with Cnp1 inactivation. AB - Fibroblast growth factor receptors (Fgfr) comprise a widely expressed family of developmental regulators implicated in oligodendrocyte (OL) maturation of the CNS. Fgfr2 is expressed by OLs in myelinated fiber tracks. In vitro, Fgfr2 is highly upregulated during OL terminal differentiation, and its activation leads to enhanced growth of OL processes and the formation of myelin-like membranes. To investigate the in vivo function of Fgfr2 signaling by myelinating glial cells, we inactivated the floxed Fgfr2 gene in mice that coexpress Cre recombinase (cre) as a knock-in gene into the OL-specific 2',3'-cyclic nucleotide phosphodiesterase (Cnp1) locus. Surprisingly, no obvious defects were detected in brain development of these conditional mutants, including the number of OLs, the onset and extent of myelination, the ultrastructure of myelin, and the expression level of myelin proteins. However, unexpectedly, a subset of these conditional Fgfr2 knock-out mice that are homozygous for cre and therefore are also Cnp1 null, displayed a dramatic hyperactive behavior starting at approximately 2 weeks of age. This hyperactivity was abolished by treatment with dopamine receptor antagonists or catecholamine biosynthesis inhibitors, suggesting that the symptoms involve a dysregulation of the dopaminergic system. Although the molecular mechanisms are presently unknown, this novel mouse model of hyperactivity demonstrates the potential involvement of OLs in neuropsychiatric disorders, as well as the nonpredictable role of genetic interactions in the behavioral phenotype of mice. PMID- 17122060 TI - Chromatic properties of horizontal and ganglion cell responses follow a dual gradient in cone opsin expression. AB - In guinea pig retina, immunostaining reveals a dual gradient of opsins: cones expressing opsin sensitive to medium wavelengths (M) predominate in the upper retina, whereas cones expressing opsin sensitive to shorter wavelengths (S) predominate in the lower retina. Whether these gradients correspond to functional gradients in postreceptoral neurons is essentially unknown. Using monochromatic flashes, we measured the relative weights with which M, S, and rod signals contribute to horizontal cell responses. For a background that produced 4.76 log10 photoisomerizations per rod per second (Rh*/rod/s), mean weights in superior retina were 52% (M), 2% (S), and 46% (rod). Mean weights in inferior retina were 9% (M), 50% (S), and 41% (rod). In superior retina, cone opsin weights agreed quantitatively with relative pigment density estimates from immunostaining. In inferior retina, cone opsin weights agreed qualitatively with relative pigment density estimates, but quantitative comparison was impossible because individual cones coexpress both opsins to varying and unquantifiable degrees. We further characterized the functional gradients in horizontal and brisk-transient ganglion cells using flickering stimuli produced by various mixtures of blue and green primary lights. Cone weights for both cell types resembled those obtained for horizontal cells using monochromatic flashes. Because the brisk-transient ganglion cell is thought to mediate behavioral detection of luminance contrast, our results are consistent with the hypothesis that the dual gradient of cone opsins assists achromatic contrast detection against different spectral backgrounds. In our preparation, rod responses did not completely saturate, even at background light levels typical of outdoor sunlight (5.14 log10 Rh*/rod/s). PMID- 17122061 TI - Biophysical model of AMPA receptor trafficking and its regulation during long term potentiation/long-term depression. AB - AMPA receptors mediate the majority of fast excitatory synaptic transmission in the CNS, and evidence suggests that AMPA receptor trafficking regulates synaptic strength, a phenomenon implicated in learning and memory. There are two major mechanisms of AMPA receptor trafficking: exocytic/endocytic exchange of surface receptors with intracellular receptor pools, and the lateral diffusion or hopping of surface receptors between the postsynaptic density and the surrounding extrasynaptic membrane. In this paper, we present a biophysical model of these trafficking mechanisms under basal conditions and during the expression of long term potentiation (LTP) and depression (LTD). We show how our model reproduces a wide range of physiological data, and use this to make predictions regarding possible targets of second-messenger pathways activated during the induction phase of LTP/LTD. PMID- 17122062 TI - The rewards of nicotine: regulation by tissue plasminogen activator-plasmin system through protease activated receptor-1. AB - Nicotine, a primary component of tobacco, is one of the most abused drugs worldwide. Approximately four million people die each year because of diseases associated with tobacco smoking. Mesolimbic dopaminergic neurons mediate the rewarding effects of abused drugs, including nicotine. Here we show that the tissue plasminogen activator (tPA)-plasmin system regulates nicotine-induced reward and dopamine release by activating protease activated receptor-1 (PAR1). In vivo microdialysis revealed that microinjection of either tPA or plasmin into the nucleus accumbens (NAc) significantly potentiated whereas plasminogen activator inhibitor-1 reduced the nicotine-induced dopamine release in the NAc in a dose-dependent manner. Nicotine-induced dopamine release was markedly diminished in tPA-deficient (tPA-/-) mice, and the defect of dopamine release in tPA-/- mice was restored by microinjection of either exogenous tPA or plasmin into the NAc. Nicotine increased tPA protein levels and promoted the release of tPA into the extracellular space in the NAc. Immunohistochemistry revealed that PAR1 immunoreactivity was localized to the nerve terminals positive for tyrosine hydroxylase in the NAc. Furthermore, we demonstrated that plasmin activated PAR1 and that nicotine-induced place preference and dopamine release were diminished in PAR1-deficient (PAR1-/-) mice. Targeting the tPA-plasmin-PAR1 system would provide new therapeutic approaches to the treatment of nicotine dependence. PMID- 17122063 TI - Time-dependent translational response of E. coli to excess Zn(II). AB - Zinc homeostasis is not well understood beyond methods of import and export. In order to better understand zinc homeostasis in Escherichia coli by identifying Zn(II)-responsive proteins, a proteomic approach was taken. Through the use of two-dimensional gel electrophoresis, we were able to show that the levels of OmpF, AspC, YcdO, Eno, and CysE increased after 30 min of Zn(II) stress, while the levels of Tig, TufA, SelA, and LeuC decreased relative to non-stressed controls. After 4 h of Zn(II) stress, the levels of three proteins (DnaK, YeaU, and Mdh) were found to be up-regulated, while the levels of seven amino acid importers (HisJ, ArgT, LivJ, DppA, OppA, RbsB, and GinH) were found to be decreased. None of these proteins had been reported to be up- or down-regulated in any previously published cDNA microarray experiments. This result raises questions about the validity of cDNA arrays when they are used to make assumptions concerning protein levels within bacterial cells. These data also suggest that time is a factor when characterizing how the E. coli proteome responds to Zn(II) stress. PMID- 17122064 TI - ABRF ESRG 2005 study: identification of seven modified amino acids by Edman sequencing. AB - Identification of modified amino acids can be a challenging part for Edman degradation sequence analysis, largely because they are not included among the commonly used phenylthiohydantion amino acid standards. Yet many can have unique retention times and can be assigned by an experienced researcher or through the use of a guide showing their typical chromatography characteristics. The Edman Sequencing Research Group (ESRG) 2005 study is a continuation of the 2004 study, in which the participating laboratories were provided a synthetic peptide and asked to identify the modified amino acids present in the sequence. The study sample provided an opportunity to sequence a peptide containing a variety of modified amino acids and note their retention times relative to the common amino acids. It also allowed the ESRG to compile the chromatographic properties and intensities from multiple instruments and tabulate an average elution position for these modified amino acids on commonly used instruments. Participating laboratories were given 2000 pmoles of a synthetic peptide, 18 amino acids long, containing the following modified amino acids: dimethyl- and trimethyl-lysine, 3 methyl-histidine, N-carbamyl-lysine, cystine, N-methyl-alanine, and isoaspartic acid. The modified amino acids were interspersed with standard amino acids to help in the assessment of initial and repetitive yields. In addition to filling in an assignment sheet, which included retention times and peak areas, participants were asked to provide specific details about the parameters used for the sequencing run. References for some of the modified amino acid elution characteristics were provided and the participants had the option of viewing a list of the modified amino acids present in the peptide at the ESRG Web site. The ABRF ESRG 2005 sample is the seventeenth in a series of studies designed to aid laboratories in evaluating their abilities to obtain and interpret amino acid sequence data. PMID- 17122065 TI - Verification of single-peptide protein identifications by the application of complementary database search algorithms. AB - Data produced from the MudPIT analysis of yeast (S. cerevisiae) and rice (O. sativa) were used to develop a technique to validate single-peptide protein identifications using complementary database search algorithms. This results in a considerable reduction of overall false-positive rates for protein identifications; the overall false discovery rates in yeast are reduced from near 25% to less than 1%, and the false discovery rate of yeast single-peptide protein identifications becomes negligible. This technique can be employed by laboratories utilizing a SEQUEST-based proteomic analysis platform, incorporating the XTandem algorithm as a complementary tool for verification of single-peptide protein identifications. We have achieved this using open-source software, including several data-manipulation software tools developed in our laboratory, which are freely available to download. PMID- 17122066 TI - Rapid and efficient MALDI-TOF MS peak detection of 2-nitrobenzenesulfenyl-labeled peptides using the combination of HPLC and an automatic spotting apparatus. AB - In this paper, we report MALDI-TOF ms analysis of 2-nitrobenzenesulfenyl (NBS) labeled peptides with the powerful aid of an LC-automatic spotting system. using this approach we analyzed mammalian sera (rat and mouse) as biological samples to demonstrate performance. The labeling was carried out using a binary set of 2 nitrobenzenesulfenyl chloride (heavy and light), which modified tryptophan residues in sample proteins. Approximately 1600 doublet peaks were detected in the mass spectrum, some of which had more than threefold differences in their intensities. systematic separation/spotting followed by mass analysis of the NBS labeled peptides derived from biological samples is described for the first time. This method has proved to be an effective application of NBS-labeled peptides and can be a powerful technique for quantitative analysis of proteins expressed in biological systems. PMID- 17122067 TI - Arabidopsis NRP1 and NRP2 encode histone chaperones and are required for maintaining postembryonic root growth. AB - NUCLEOSOME ASSEMBLY PROTEIN1 (NAP1) is conserved from yeast to human and was proposed to act as a histone chaperone. While budding yeast contains a single NAP1 gene, multicellular organisms, including plants and animals, contain several NAP1 and NAP1-RELATED PROTEIN (NRP) genes. However, the biological role of these genes has been largely unexamined. Here, we show that, in Arabidopsis thaliana, simultaneous knockout of the two NRP genes, NRP1 and NRP2, impaired postembryonic root growth. In the nrp1-1 nrp2-1 double mutant, arrest of cell cycle progression at G2/M and disordered cellular organization occurred in root tips. The mutant seedlings exhibit perturbed expression of approximately 100 genes, including some genes involved in root proliferation and patterning. The mutant plants are highly sensitive to genotoxic stress and show increased levels of DNA damage and the release of transcriptional gene silencing. NRP1 and NRP2 are localized in the nucleus and can form homomeric and heteromeric protein complexes. Both proteins specifically bind histones H2A and H2B and associate with chromatin in vivo. We propose that NRP1 and NRP2 act as H2A/H2B chaperones in the maintenance of dynamic chromatin in epigenetic inheritance. PMID- 17122068 TI - Arabidopsis CUP-SHAPED COTYLEDON3 regulates postembryonic shoot meristem and organ boundary formation. AB - Overall shoot architecture in higher plants is highly dependent on the activity of embryonic and axillary shoot meristems, which are produced from the basal adaxial boundaries of cotyledons and leaves, respectively. In Arabidopsis thaliana, redundant functions of the CUP-SHAPED COTYLEDON genes CUC1, CUC2, and CUC3 regulate embryonic shoot meristem formation and cotyledon boundary specification. Their functional importance and relationship in postembryonic development, however, is poorly understood. Here, we performed extensive analyses of the embryonic and postembryonic functions of the three CUC genes using multiple combinations of newly isolated mutant alleles. We found significant roles of CUC2 and CUC3, but not CUC1, in axillary meristem formation and boundary specification of various postembryonic shoot organs, such as leaves, stems, and pedicels. In embryogenesis, all three genes make significant contributions, although CUC3 appears to possess, at least partially, a distinct function from that of CUC1 and CUC2. The function of CUC3 and CUC2 overlaps that of LATERAL SUPPRESSOR, which was previously shown to be required for axillary meristem formation. Our results reveal that redundant but partially distinct functions of CUC1, CUC2, and CUC3 are responsible for shoot organ boundary and meristem formation throughout the life cycle in Arabidopsis. PMID- 17122069 TI - The Arabidopsis histidine phosphotransfer proteins are redundant positive regulators of cytokinin signaling. AB - Arabidopsis thaliana histidine phosphotransfer proteins (AHPs) are similar to bacterial and yeast histidine phosphotransfer proteins (HPts), which act in multistep phosphorelay signaling pathways. A phosphorelay pathway is the current model for cytokinin signaling. To assess the role of AHPs in cytokinin signaling, we isolated T-DNA insertions in the five AHP genes that are predicted to encode functional HPts and constructed multiple insertion mutants, including an ahp1,2,3,4,5 quintuple mutant. Single ahp mutants were indistinguishable from wild-type seedlings in cytokinin response assays. However, various higher-order mutants displayed reduced sensitivity to cytokinin in diverse cytokinin assays, indicating both a positive role for AHPs in cytokinin signaling and functional overlap among the AHPs. In contrast with the other four AHPs, AHP4 may play a negative role in some cytokinin responses. The quintuple ahp mutant showed various abnormalities in growth and development, including reduced fertility, increased seed size, reduced vascular development, and a shortened primary root. These data indicate that most of the AHPs are redundant, positive regulators of cytokinin signaling and affect multiple aspects of plant development. PMID- 17122070 TI - Aminocyclopropane carboxylic acid synthase is a regulated step in ethylene dependent induced conifer defense. Full-length cDNA cloning of a multigene family, differential constitutive, and wound- and insect-induced expression, and cellular and subcellular localization in spruce and Douglas fir. AB - In conifer stems, formation of chemical defenses against insects or pathogens involves specialized anatomical structures of the phloem and xylem. Oleoresin terpenoids are formed in resin duct epithelial cells and phenolics accumulate in polyphenolic parenchyma cells. Ethylene signaling has been implicated in the induction of these chemical defenses. Recently, we reported the cloning of 1 aminocyclopropane-1-carboxylic acid oxidase (ACO) from spruce (Picea spp.) and Douglas fir (Pseudotsuga menziesii). ACO protein was constitutively expressed in Douglas fir and only weakly induced upon wounding. We now cloned seven full length and one near full-length cDNA representing four distinct 1 aminocyclopropane-1-carboxylic acid synthases (ACS; ACS1, ACS2, ACS3, and ACS4) from spruce and Douglas fir. Cloning of ACS has not previously been reported for any gymnosperm. Using gene-specific, quantitative real-time polymerase chain reaction, we measured constitutive expression for the four ACS genes and the single-copy ACO gene in various tissues of Sitka spruce (Picea sitchensis) and in white spruce (Picea glauca) somatic embryos. ACO and ACS4 were ubiquitously expressed at high levels; ACS1 was predominantly expressed in developing embryos and ACS2 and ACS3 were expressed only at very low levels. Insect attack or mechanical wounding caused strong induction of ACS2 and ACS3 in Sitka spruce bark, a moderate increase in ACO transcripts, but had no effect on ACS1 and ACS4. ACS protein was also strongly induced following mechanical wounding in Douglas fir and was highly abundant in resin duct epithelial cells and polyphenolic parenchyma cells. These results suggest that ACS, but not ACO, is a regulated step in ethylene-induced conifer defense. PMID- 17122071 TI - Circadian rhythms of isoprene biosynthesis in grey poplar leaves. AB - Isoprene (2-methyl-1,3-butadiene) emission varies diurnally in different species. In poplar (Populus spp.), it has recently been shown that the gene encoding the synthesizing enzyme for isoprene, isoprene synthase (ISPS), displays diurnal variation in expression. Working on shoot cultures of Grey poplar (Populus x canescens) placed under a different light regime in phytochambers, we showed that these variations in PcISPS gene expression, measured by quantitative real-time polymerase chain reaction, are not only due to day-night changes, but also are linked to an internal circadian clock. Measurement of additional selected isoprenoid genes revealed that phytoene synthase (carotenoid pathway) displays similar fluctuations, whereas 1-deoxy-d-xylulose 5-phosphate reductoisomerase, possibly the first committed enzyme of the 1-deoxy-d-xylulose 5-phosphate pathway, only shows light regulation. On the protein level, it appeared that PcISPS activity and protein content became reduced under constant darkness, whereas under constant light, activity and protein content of this enzyme were kept high. In contrast, isoprene emission rates under continuous irradiation displayed circadian changes as is the case for gene expression of PcISPS. Furthermore, binding assays with Arabidopsis (Arabidopsis thaliana) late elongated hypocotyl, a transcription factor of Arabidopsis involved in circadian regulation, clearly revealed the presence of circadian-determining regulatory elements in the promoter region of PcISPS. PMID- 17122072 TI - The metabolic response of heterotrophic Arabidopsis cells to oxidative stress. AB - To cope with oxidative stress, the metabolic network of plant cells must be reconfigured either to bypass damaged enzymes or to support adaptive responses. To characterize the dynamics of metabolic change during oxidative stress, heterotrophic Arabidopsis (Arabidopsis thaliana) cells were treated with menadione and changes in metabolite abundance and (13)C-labeling kinetics were quantified in a time series of samples taken over a 6 h period. Oxidative stress had a profound effect on the central metabolic pathways with extensive metabolic inhibition radiating from the tricarboxylic acid cycle and including large sectors of amino acid metabolism. Sequential accumulation of metabolites in specific pathways indicated a subsequent backing up of glycolysis and a diversion of carbon into the oxidative pentose phosphate pathway. Microarray analysis revealed a coordinated transcriptomic response that represents an emergency coping strategy allowing the cell to survive the metabolic hiatus. Rather than attempt to replace inhibited enzymes, transcripts encoding these enzymes are in fact down-regulated while an antioxidant defense response is mounted. In addition, a major switch from anabolic to catabolic metabolism is signaled. Metabolism is also reconfigured to bypass damaged steps (e.g. induction of an external NADH dehydrogenase of the mitochondrial respiratory chain). The overall metabolic response of Arabidopsis cells to oxidative stress is remarkably similar to the superoxide and hydrogen peroxide stimulons of bacteria and yeast (Saccharomyces cerevisiae), suggesting that the stress regulatory and signaling pathways of plants and microbes may share common elements. PMID- 17122073 TI - Alleviation of thermoregulatory dysfunction with the new serotonin and norepinephrine reuptake inhibitor desvenlafaxine succinate in ovariectomized rodent models. AB - Hot flushes and night sweats, referred to as vasomotor symptoms (VMS), are presumed to be a result of declining hormone levels and are the principal menopausal symptoms for which women seek medical treatment. To date, estrogens and/or some progestins are the most effective therapeutics for alleviating VMS; however, these therapies may not be appropriate for all women. Therefore, nonhormonal therapies are being evaluated. The present study investigated a new reuptake inhibitor, desvenlafaxine succinate (DVS), in animal models of temperature dysfunction. Both models used are based on measuring changes in tail skin temperature (TST) in ovariectomized (OVX) rats. The first relies on naloxone induced withdrawal in morphine-dependent (MD) OVX rats, resulting in an acute rise in TST. The second depends on an OVX-induced loss of TST decreases during the dark phase as measured by telemetry. An initial evaluation demonstrated abatement of the rise in TST with long-term administration of ethinyl estradiol or with a single oral dose of DVS (130 mg/kg) in the MD model. Further evaluation showed that orally administered DVS acutely and dose dependently (10-100 mg/kg) abated a naloxone-induced rise in TST of MD rats and alleviated OVX-induced temperature dysfunction in the telemetry model. Oral administration of DVS to OVX rats caused significant increases in serotonin and norepinephrine levels in the preoptic area of the hypothalamus, a key region of the brain involved in temperature regulation. These preclinical studies provide evidence that DVS directly impacts thermoregulatory dysfunction in OVX rats and may have utility in alleviating VMS associated with menopause. PMID- 17122074 TI - Single-chain bifunctional vascular endothelial growth factor (VEGF)-follicle stimulating hormone (FSH)-C-terminal peptide (CTP) is superior to the combination therapy of recombinant VEGF plus FSH-CTP in stimulating angiogenesis during ovarian folliculogenesis. AB - Infertility technologies often employ exogenous gonadotropin therapy to increase antral follicle production. In an effort to enhance ovarian response, several long-acting FSH therapies have been developed including an FSH-C-terminal peptide (CTP), where the FSH subunits are linked by the CTP moiety from human chorionic gonadotropin, which is responsible for the increased half-life of human chorionic gonadotropin. We found that administration of FSH-CTP for ovarian hyperstimulation in rats blunted ovarian follicle vascular development. In women, reduced ovarian vasculature has been associated with lower pregnancy rates. We were interested in determining whether vascular endothelial growth factor (VEGF) therapy could enhance ovarian angiogenesis in FSH-CTP-treated rats. Coadministration of systemic FSH-CTP plus recombinant VEGF was compared with treatment with a novel, single-chain bifunctional VEGF-FSH-CTP (VFC) analog. For VFC, the FSH portion targets the protein to the ovary and stimulates follicle growth, whereas VEGF enhances local vascular development. Both in vitro and in vivo studies confirm the dual FSH and VEGF action of the VFC protein. Evaluation of ovarian follicle development demonstrates that administration of combination therapy using VEGF and FSH-CTP failed to increase follicle vasculature above levels seen with FSH-CTP monotherapy. However, treatment with VFC significantly increased follicle vascular development while concurrently increasing the number of large antral follicles produced. In conclusion, we report the production and characterization of a long-acting, bifunctional VEGF-FSH-CTP protein that is superior to combination therapy for enhancing VEGF activity in the ovary and stimulating follicular angiogenesis in rats. PMID- 17122075 TI - Adrenal 20alpha-hydroxysteroid dehydrogenase in the mouse catabolizes progesterone and 11-deoxycorticosterone and is restricted to the X-zone. AB - The enzyme 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD) is a progesterone catabolizing enzyme that is highly expressed in mouse ovaries and adrenals. Although the functional significance of ovarian 20alpha-HSD for the induction of parturition has been defined, regulation and distribution of 20alpha-HSD in the adrenal gland has not been determined. We demonstrate that the expression of adrenal 20alpha-HSD is restricted to the X-zone, a transient zone between the adrenal cortex and the medulla of yet unknown function. Adrenal 20alpha-HSD activity in male mice peaks at 3 wk of age and disappears thereafter, whereas 20alpha-HSD enzyme activity is maintained in adrenals from nulliparous female animals. Testosterone treatment of female mice induces rapid involution of the X zone that is associated with the disappearance of the 20alpha-HSD-positive cells. Conversely, reappearance of 20alpha-HSD expression and activity in male animals is evident after gonadectomy. Moreover, pregnancy, but not pseudopregnancy, is accompanied by X-zone regression and loss of 20alpha-HSD activity. Pregnancy induced X-zone regression and -abolished 20alpha-HSD expression is partially restored in animals that were kept from nursing their pups. We found that in addition to its progesterone-reducing activity, 20alpha-HSD also functions as an 11-deoxycorticosterone-catabolizing enzyme. The unaltered growth kinetics of the X-zone in 20alpha-HSD knockout animals suggests that 20alpha-HSD is not required for the regulation of X-zone growth. However, 20alpha-HSD expression and enzymatic activity in all experimental paradigms is closely correlated with the presence of the X-zone. These findings provide the basis for 20alpha-HSD as a reliable marker of the murine X-zone. PMID- 17122076 TI - Involvement of cholecystokinin 2 receptor in food intake regulation: hyperphagia and increased fat deposition in cholecystokinin 2 receptor-deficient mice. AB - The role of cholecystokinin (CCK) as a satiety factor has been extensively documented. Although most work implies that CCK1 receptor mediates the control of food intake, a contributing role for CCK2 receptor (CCK2R) in the CCK-induced satiety cannot be totally excluded. The hypothesis that CCK2R invalidation disrupts regulatory pathways with impact on feeding behavior was examined in CCK2R(-/-) mice. CCK2R(-/-) mice developed obesity that was associated with hyperphagia. Obesity was related with increased fat deposition resulting from adipocyte hypertrophy. Expression of several adipokines was dysregulated consistently with obesity. Moreover, obesity was associated with disturbed glucose homeostasis as revealed by increased fasting glycemia and insulinemia, impaired glucose tolerance, and hepatic insulin resistance in CCK2R(-/-) mice. In vitro analysis of isolated adipocytes metabolism was consistent with increased storage but preserved insulin sensitivity. Suppression of feeding and concomitant increased expression of hypothalamic proopiomelanocortin after intracerebroventricular injection of gastrin into control mice demonstrates that hypothalamic CCK2 receptors mediate inhibition of food intake. Comparative analysis of hypothalamic mediator gene expression in fed knockout and control mice demonstrated overexpression of ghrelin receptors in CCK2R(-/-) mice, indicating up-regulation of orexigenic pathways. This effect was also observed after body weight normalization, indicating a causative role in the development of hyperphagia and obesity of CCK2R(-/-) mice. Our results give evidence that CCK2 receptor activity plays a contributing regulatory role in the control of food intake. PMID- 17122077 TI - Inositide-dependent phospholipase C signaling mimics insulin in skeletal muscle differentiation by affecting specific regions of the cyclin D3 promoter. AB - Our main goal in this study was to investigate the role of phospholipase C (PLC) beta(1) and PLCgamma(1) in skeletal muscle differentiation and the existence of potential downstream targets of their signaling activity. To examine whether PLC signaling can modulate the expression of cyclin D3, a target of PLCbeta(1) in erythroleukemia cells, we transfected C2C12 cells with expression vectors containing PLCbeta(1) or PLCgamma(1) cDNA and with small interfering RNAs from regions of the PLCbeta(1) or PLCgamma(1) gene and followed myogenic differentiation in this well-established cell system. Intriguingly, overexpressed PLCbeta(1) and PLCgamma(1) were able to mimic insulin induction of both cyclin D3 and muscle differentiation. By knocking down PLCbeta(1) or PLCgamma(1) expression, C2C12 cells almost completely lost the increase in cyclin D3, and the differentiation program was down-regulated. To explore the induction of the cyclin D3 gene promoter during this process, we used a series of 5'-deletions of the 1.68-kb promoter linked to a reporter gene and noted a 5-fold augmentation of promoter activity upon insulin stimulation. These constructs were also cotransfected with PLCbeta(1) or PLCgamma(1) cDNAs and small interfering RNAs, respectively. Our data indicate that PLCbeta(1) or PLCgamma(1) signaling is capable of acting like insulin in regard to both the myogenic differentiation program and cyclin D3 up-regulation. Taken together, this is the first study that hints at cyclin D3 as a target of PLCbeta(1) and PLCgamma(1) during myogenic differentiation in vitro and implies that up-regulation of these enzymes is sufficient to mimic the actions of insulin in this process. PMID- 17122078 TI - A mutant thyroid hormone receptor alpha antagonizes peroxisome proliferator activated receptor alpha signaling in vivo and impairs fatty acid oxidation. AB - Thyroid hormone regulates the balance between lipolysis and lipogenesis. We previously reported that male mice with a dominant-negative P398H mutation introduced into the TRalpha gene have visceral obesity, hyperleptinemia, and reduced catecholamine-stimulated lipolysis in white adipose tissue. Based on our observation of hepatic steatosis in the TRalpha P398H male mice, we used in vitro and in vivo models to investigate the influence of the TRalpha P398H mutant on peroxisome proliferator-activated receptor-alpha (PPARalpha) signaling. Wild-type TRalpha and the P398H mutant significantly reduced PPARalpha-mediated transcription in transient transfection assays. T(3) reversed the inhibition of PPARalpha action by wild-type TRalpha but not the P398H mutant. Chromatin immunoprecipitation assays demonstrated that the P398H mutant reduces PPARalpha binding to peroxisome proliferator receptor elements. In gel shift assays, the P398H mutant directly bound the peroxisome proliferator-activated receptor response element and inhibited PPARalpha binding, which was not reversed by addition of retinoid X receptor. The TRalpha R384C and PV dominant-negative mutants are not associated in vivo with a metabolic phenotype and had reduced (PV) or absent (R384C) PPARalpha inhibition compared with P398H. The metabolic phenotype of the P398H mutant mice is due, in part, to unique properties of the P398H mutant receptor interfering with PPARalpha signaling. The P398H mutant is a potential probe to characterize the physiological role of thyroid hormone receptor/PPARalpha interactions. PMID- 17122079 TI - Functional corpora lutea are formed in matrix metalloproteinase inhibitor-treated plasminogen-deficient mice. AB - Corpus luteum (CL) formation involves dramatic tissue remodeling and angiogenesis. To determine the functional roles of the plasminogen activator and matrix metalloproteinase (MMP) systems in these processes, we have studied CL formation and function in plasminogen (plg)-deficient mice, with or without treatment with the broad-spectrum synthetic MMP inhibitor galardin. Both the adult pseudopregnant CL model and the gonadotropin-primed immature mouse model were used. We found that CL formed normally not only in plasminogen-deficient mice and in galardin-treated wild-type mice, but also in galardin-treated plg deficient mice, suggesting that neither of the plasminogen activator and MMP systems is essential for CL formation. Nevertheless, in plg-deficient mice, serum progesterone levels were reduced by approximately 50%, and the progesterone levels were not reduced further by galardin treatment. When CL from plg-deficient mice were stained for several molecular markers for CL development and regression, they appeared healthy and vascularized, and were indistinguishable from CL from wild-type mice. This implies that the reduced progesterone levels were not caused by impaired CL formation. Taken together, our data suggest that neither plasmin nor MMPs, alone or in combination, are required for CL formation. Therefore, the tissue remodeling and angiogenesis processes during CL formation may be mediated by redundant protease systems. However, the reduced serum progesterone levels in plg-deficient mice suggest that plasmin, but not MMPs, plays a role in maintenance of luteal function. This role may be performed through proteolytic activation of growth factors and other paracrine factors. PMID- 17122080 TI - C-terminal fragments of the gastrin-releasing peptide precursor stimulate cell proliferation via a novel receptor. AB - There are many precedents for the production from a single precursor of multiple peptides, with independent receptors and different bioactivities. Gastrin releasing peptide (GRP) is initially synthesized as amino acids 1-27 of a 125 residue precursor, proGRP, and is subsequently cleaved and amidated to form GRP18 27. We investigated the hypothesis that C-terminal proGRP peptides are also biologically active. Human proGRP18-125 was expressed in Escherichia coli as a glutathione S-transferase fusion protein. Recombinant proGRP18-125 stimulated proliferation and migration of the human colorectal carcinoma cell line DLD-1. The observations that an antagonist selective for the GRP receptor did not inhibit activity in either proliferation or migration assays and that the recombinant peptide did not bind to either the GRP receptor or orphan receptor BRS-3 indicated that neither activity was mediated by the known GRP receptors. Recombinant human proGRP31-125 and proGRP42-98 were also prepared and shown to stimulate proliferation of DLD-1 cells and the human prostate carcinoma cell line DU145. The synthetic peptides proGRP47-68 and [Tyr79]proGRP80-97 stimulated inositol phosphate production, MAPK kinase activity, and proliferation and migration of DLD-1 cells. Binding sites for both radioiodinated synthetic peptides were demonstrated on DLD-1 cells. Each peptide was able to compete with the other for binding, and a GRP receptor antagonist did not inhibit binding of either peptide. We conclude that peptides derived from the C terminus of proGRP are biologically active and that their activity is mediated by a receptor distinct from the two known GRP receptors. PMID- 17122081 TI - The hypothalamic-pituitary-adrenal axis response to stress in mice lacking functional vasopressin V1b receptors. AB - The role of arginine vasopressin (Avp) as an ACTH secretagogue is mediated by the Avp 1b receptor (Avpr1b) found on anterior pituitary corticotropes. Avp also potentiates the actions of CRH (Crh) and appears to be an important mediator of the hypothalamic-pituitary-adrenal axis response to chronic stress. To investigate the role of Avp in the hypothalamic-pituitary-adrenal axis response to stress, we measured plasma ACTH and corticosterone (CORT) levels in Avpr1b knockout (KO) mice and wild-type controls in response to two acute (restraint and insulin administration) and one form of chronic (daily restraint for 14 d) stress. No significant difference was found in the basal plasma levels of ACTH and CORT between the two genotypes. Acute restraint (30 min) increased plasma ACTH and CORT to a similar level in both the Avpr1b mutant and wild-type mice. In contrast, plasma ACTH and CORT levels induced by hypoglycemia were significantly decreased in the Avpr1b KO mice when compared with wild-type littermates. There was no difference in the ACTH response to acute and chronic restraint in wild type mice. In the Avpr1b KO group subjected to 14 sessions of daily restraint, plasma ACTH was decreased when compared with wild-type mice. On the other hand, the CORT elevations induced by restraint did not adapt in the Avpr1b KO or wild type mice. The data suggest that the Avpr1b is required for the normal pituitary and adrenal response to some acute stressful stimuli and is necessary only for a normal ACTH response during chronic stress. PMID- 17122082 TI - Estradiol induces expression of 5-hydroxytryptamine (5-HT) 4, 5-HT5, and 5-HT6 receptor messenger ribonucleic acid in rat anterior pituitary cell aggregates and allows prolactin release via the 5-HT4 receptor. AB - Serotonin [5-hydroxytryptamine (5-HT)] is known to control prolactin (PRL) release at a hypothalamic level, but a pituitary site of action remains poorly studied. The present study explores the acute effect of 5-HT on PRL release in rat anterior pituitary aggregate cell cultures, the influence of steroid and thyroid hormones, and the 5-HT receptor (5-HTR) subtype(s) involved. 5-HT elicited a prompt increase in basal PRL release, an effect strongly potentiated by estradiol (E(2)) in the culture medium (dose response 1-100 nm). In E(2) condition, the PRL response was not affected by the nonselective 5-HTR antagonists methysergide and methiothepin nor by 5-HTR1, 5-HTR2, 5-HTR3, 5-HTR6, and 5-HTR7/5 antagonists, but was fully blocked by the 5-HTR4 antagonist GR 113808. Among various agonist analogs, only the 5-HTR4 agonist cisapride and the 5-HTR2 agonist alpha-methyl-5-HT evoked PRL release. The effect of alpha-methyl-5 HT also required E(2) during culture and was abolished by GR 113808 but not by combined 5-HTR2A, B, and C blockade. In E(2)-treated aggregates, 5-HT caused a 5 fold increase in cAMP levels. The intact anterior pituitary expressed mRNA of all known members of the 5-HTR family. In aggregates, 5-HTR4, 5-HTR5, and 5-HTR6 mRNA expression required E(2) during culture. The effect of 5-HT on PRL release was not affected by blocking the serotonin transporter or the vesicular monoamine transporter. The present data suggest a widespread expression of 5-HTRs in the rat anterior pituitary, several of which are up-regulated by estrogen, and that, in the presence of estrogen, one of these, the 5-HTR4, mediates acute PRL release. PMID- 17122083 TI - Activation of peroxisome proliferator-activated receptor gamma (PPARgamma) by rosiglitazone suppresses components of the insulin-like growth factor regulatory system in vitro and in vivo. AB - Rosiglitazone (Rosi) belongs to the class of thiazolidinediones (TZDs) that are ligands for peroxisome proliferator-activated receptor gamma (PPARgamma). Stimulation of PPARgamma suppresses bone formation and enhances marrow adipogenesis. We hypothesized that activation of PPARgamma down-regulates components of the IGF regulatory system, leading to impaired osteoblast function. Rosi treatment (1 microm) of a marrow stromal cell line (UAMS-33) transfected with empty vector (U-33/c) or with PPARgamma2 (U-33/gamma2) were analyzed by microarray. Rosi reduced IGF-I, IGF-II, IGFBP-4, and the type I and II IGF receptor (IGF1R and IGF2R) expression at 72 h in U-33/gamma2 compared with U-33/c cells (P < 0.01); these findings were confirmed by RT-PCR. Rosi reduced secreted IGF-I from U-33/gamma2 cells by 75% (P < 0.05). Primary marrow stromal cells (MSCs) extracted from adult (8 months) and old (24 months) C57BL/6J (B6) mice were treated with Rosi (1 microm) for 48 h. IGF-I, IGFBP-4, and IGF1R transcripts were reduced in Rosi-treated MSCs compared with vehicle (P < 0.01) and secreted IGF-I was also suppressed (P < 0.05). B6 mice treated with Rosi (20 mg/kg.d) for short duration (i.e. 4 d), and long term (i.e. 7 wk) had reduced serum IGF-I; this was accompanied by markedly suppressed IGF-I transcripts in the liver and peripheral fat of treated animals. To determine whether Rosi affected circulating IGF-I in humans, we measured serum IGF-I, IGFBP-2, and IGFBP-3 at four time points in 50 postmenopausal women randomized to either Rosi (8 mg/d) or placebo. Rosi-treated subjects had significantly lower IGF-I at 8 wk than baseline (-25%, P < 0.05), and at 16 wk their levels were reduced 14% vs. placebo (P = 0.15). We conclude that Rosi suppresses IGF-I expression in bone and liver; these changes could affect skeletal acquisition through endocrine and paracrine pathways. PMID- 17122084 TI - Genome-wide detection of human copy number variations using high-density DNA oligonucleotide arrays. AB - Recent reports indicate that copy number variations (CNVs) within the human genome contribute to nucleotide diversity to a larger extent than single nucleotide polymorphisms (SNPs). In addition, the contribution of CNVs to human disease susceptibility may be greater than previously expected, although a complete understanding of the phenotypic consequences of CNVs is incomplete. We have recently reported a comprehensive view of CNVs among 270 HapMap samples using high-density SNP genotyping arrays and BAC array CGH. In this report, we describe a novel algorithm using Affymetrix GeneChip Human Mapping 500K Early Access (500K EA) arrays that identified 1203 CNVs ranging in size from 960 bp to 3.4 Mb. The algorithm consists of three steps: (1) Intensity pre-processing to improve the resolution between pairwise comparisons by directly estimating the allele-specific affinity as well as to reduce signal noise by incorporating probe and target sequence characteristics via an improved version of the Genomic Imbalance Map (GIM) algorithm; (2) CNV extraction using an adapted SW-ARRAY procedure to automatically and robustly detect candidate CNV regions; and (3) copy number inference in which all pairwise comparisons are summarized to more precisely define CNV boundaries and accurately estimate CNV copy number. Independent testing of a subset of CNVs by quantitative PCR and mass spectrometry demonstrated a >90% verification rate. The use of high-resolution oligonucleotide arrays relative to other methods may allow more precise boundary information to be extracted, thereby enabling a more accurate analysis of the relationship between CNVs and other genomic features. PMID- 17122085 TI - Accurate and reliable high-throughput detection of copy number variation in the human genome. AB - This study describes a new tool for accurate and reliable high-throughput detection of copy number variation in the human genome. We have constructed a large-insert clone DNA microarray covering the entire human genome in tiling path resolution that we have used to identify copy number variation in human populations. Crucial to this study has been the development of a robust array platform and analytic process for the automated identification of copy number variants (CNVs). The array consists of 26,574 clones covering 93.7% of euchromatic regions. Clones were selected primarily from the published "Golden Path," and mapping was confirmed by fingerprinting and BAC-end sequencing. Array performance was extensively tested by a series of validation assays. These included determining the hybridization characteristics of each individual clone on the array by chromosome-specific add-in experiments. Estimation of data reproducibility and false-positive/negative rates was carried out using self-self hybridizations, replicate experiments, and independent validations of CNVs. Based on these studies, we developed a variance-based automatic copy number detection analysis process (CNVfinder) and have demonstrated its robustness by comparison with the SW-ARRAY method. PMID- 17122086 TI - Inhibitory role of Src family tyrosine kinases on Ca2+-dependent insulin release. AB - Both neurotransmitter release and insulin secretion occur via regulated exocytosis and share a variety of similar regulatory mechanisms. It has been suggested that Src family tyrosine kinases inhibit neurotransmitter release from neuronal cells (H. Ohnishi, S. Yamamori, K. Ono, K. Aoyagi, S. Kondo, and M. Takahashi. Proc Natl Acad Sci USA 98: 10930-10935, 2001). Thus the potential role of Src family kinases in the regulation of insulin secretion was investigated in this study. Two structurally different inhibitors of Src family kinases, SU-6656 and PP2, but not the inactive compound, PP3, enhanced Ca2+-induced insulin secretion in both rat pancreatic islets and INS-1 cells in a concentration dependent and time-dependent manner. Furthermore, Src family kinase-mediated insulin secretion appears to be dependent on elevated intracellular Ca2+ and independent of glucose metabolism, the ATP-dependent K+ channel, adenylyl cyclase, classical PKC isoforms, extracellular signal-regulated kinase 1/2, and insulin synthesis. The sites of action for Src family kinases seem to be distal to the elevation of intracellular Ca2+ level. These results indicate that one or more Src family tyrosine kinases exert a tonic inhibitory role on Ca2+-dependent insulin secretion. PMID- 17122087 TI - Glucose-induced release of nitric oxide from mouse pancreatic islets as detected with nitric oxide-selective glass microelectrodes. AB - Nitric oxide (NO) is believed to play an important role in pancreatic islet physiology and pathophysiology. Research in this area has been hampered, however, by the use of indirect methods to measure islet NO. To investigate the role of NO in islet function, we positioned NO-sensitive, recessed-tip microelectrodes in close proximity to individual islets and monitored oxidation current to detect subnanomolar NO in the bath. NO release from islets consisted of a series of rapid bursts lasting several seconds and/or slow oscillations with a period of approximately 100-300 s. Average baseline NO near the islets in 2.8 mM glucose was 524+/-59 nM (n=12). Raising glucose from 2.8 to 11.1 mM augmented NO release by 429+/-133 nM (n=12, P<0.05), an effect blocked by the NO synthase inhibitor L NAME (n=3). We also observed that glucose-stimulated increases in NO release were contemporaneous with changes in NAD(P)H and O2 but occurred well before increases in calcium associated with glucose-stimulated insulin secretion. In summary, we demonstrate that NO release from islets is oscillatory and rapidly augmented by glucose, suggesting that NO release occurs early following an increase in glucose metabolism and may contribute to the stimulated insulin secretion triggered by suprathreshold glucose. PMID- 17122088 TI - Impact of proestrous milieu on expression of orexin receptors and prepro-orexin in rat hypothalamus and hypophysis: actions of Cetrorelix and Nembutal. AB - Orexins and their receptors OX1 and OX2 regulate energy balance and the sleep wake cycle. We studied the expression of prepro-orexin (PPO), OX1, and OX2 in brain and pituitary under the influence of the hormonal status in adult rats. Primarily, PPO, OX1, and OX2 expression was determined in Sprague-Dawley female cycling rats during proestrus and in males. Animals were killed at 2-h intervals. Anterior (AH) and mediobasal (MBH) hypothalamus, anterior pituitary (P), and frontoparietal cortex (CC) were homogenized in TRIzol, and mRNAs were obtained for screening of PPO, OX1, OX2 expression by semiquantitative RT-PCR. Main findings were confirmed and extended to all days of the cycle by quantitative real-time RT-PCR. Hormones and food consumption were determined. Finally, OX1, OX2, and PPO were measured by real-time RT-PCR in tissues collected at 1900 of proestrus after treatments at 1400 with ovulation-blocking agents Cetrorelix or pentobarbital. OX1 and OX2 expression increased at least threefold in AH, MBH, and P, but not in CC, between 1700 and 2300 of proestrus, without variations in estrus, diestrus, or in males. PPO in AH and MBH showed a fourfold or higher increase only during proestrus afternoon. Cetrorelix or pentobarbital prevented increases of OX1 and OX2 only in the pituitary and blunted gonadotropin surges, but left OX1, OX2, and PPO brain expression unchanged. Reproduction, energy balance, and sleep-wake cycle are integrated. Here, we demonstrate that, in the physiological neuroendocrine condition leading to ovulation, information to the orexinergic system acts in hypothalamus and pituitary by different mechanisms. PMID- 17122089 TI - Acute and chronic regulation of leptin synthesis, storage, and secretion by insulin and dexamethasone in human adipose tissue. AB - Serum leptin levels are upregulated in proportion to body fat and also increase over the short term in response to meals or insulin. To understand the mechanisms involved, we assessed leptin synthesis and secretion in samples of adipose tissue from subjects with a wide range of BMI. Tissue leptin content and relative rates of leptin biosynthesis, as determined by metabolic labeling, were highly correlated with each other and with BMI and fat cell size. To understand mechanisms regulating leptin synthesis in obesity, we used biosynthetic labeling to directly assess the effects of insulin and glucocorticoids (dexamethasone) on leptin synthesis and secretion in human adipose tissue. Chronic treatment (1-2 days in organ culture) with insulin increased relative rates of leptin biosynthesis without affecting leptin mRNA levels. In contrast, dexamethasone increased leptin mRNA and biosynthesis in parallel. Acute treatment with insulin or dexamethasone (added during 1-h preincubation and 45-min pulse labeling) did not affect relative rates of leptin biosynthesis, but pulse-chase studies showed that addition of insulin nearly doubled the release of [35S]leptin after a 1-h chase. We conclude that the higher leptin stores in adipose tissue of obese humans are maintained by chronic effects of insulin and glucocorticoids acting at pre- and posttranslational levels and that the ability of insulin to increase the release of preformed leptin may contribute to short-term variations in circulating leptin levels. PMID- 17122090 TI - Progesterone acutely increases LH pulse amplitude but does not acutely influence nocturnal LH pulse frequency slowing during the late follicular phase in women. AB - Progesterone (P) is the primary effector of LH (and by inference gonadotropin releasing hormone) pulse frequency slowing in cycling women, but the time course of this action is unclear. We hypothesized that P administration to estradiol (E2)-pretreated women would slow LH pulse frequency within 12 h. We studied eight normally cycling women in two separate cycles (follicular phase, cycle days 7 11). After 3 days of E2 pretreatment (0.2 mg/day via transdermal patches), a 25-h blood sampling protocol (starting at 0800) was performed to define LH pulsatility. Oral micronized P (100 mg) or placebo (PBO) was administered at 1800 in a randomized, double-blind fashion, with treatment crossover occurring during a subsequent cycle. The 10-h mean P concentration increased from 0.6+/-0.1 ng/ml before P (0800-1800) to 3.9+/-0.3 ng/ml after P administration (2200-0800, P<0.01). Ten-hour mean LH interpulse interval increased significantly after both P and PBO administration, with no significant difference between P and PBO. In contrast, mean LH, LH amplitude, and mean FSH increased significantly within 4 h of P administration, but not after PBO. We conclude that, in E2-pretreated women in the late follicular phase, 1) nocturnal LH pulse frequency is not acutely (within 12 h) influenced by P administration; 2) an acute increase in P causes pronounced augmentation of gonadotropin pulse amplitude within 4 h; and 3) LH pulse frequency slows overnight during the second half of the follicular phase. PMID- 17122091 TI - Effects of leptin replacement on hypothalamic-pituitary growth hormone axis function and circulating ghrelin levels in ob/ob mice. AB - Leptin-deficient obese mice (ob/ob) have decreased circulating growth hormone (GH) and pituitary GH and ghrelin receptor (GHS-R) mRNA levels, whereas hypothalamic GH-releasing hormone (GHRH) and somatostatin (SST) expression do not differ from lean controls. Given the fact that GH is suppressed in diet-induced obesity (a state of hyperleptinemia), it remains to be determined whether the absence of leptin contributes to changes in the GH axis of ob/ob mice. Therefore, to study the impact of leptin replacement on the hypothalamic-pituitary GH axis of ob/ob mice, leptin was infused for 7 days (sc), resulting in circulating leptin levels that were similar to wild-type controls (approximately 1 ng/ml). Leptin treatment reduced food intake, body weight, and circulating insulin while elevating circulating n-octanoyl ghrelin concentrations. Leptin treatment did not alter hypothalamic GHRH, SST, or GHS-R mRNA levels compared with vehicle-treated controls. However, leptin significantly increased pituitary GH and GHRH-R expression and tended to enhance circulating GH levels, but this latter effect did not reach statistical significance. In vitro, leptin (1 ng/ml, 24 h) did not affect pituitary GH, GHRH-R, or GHS-R mRNA but did enhance GH release. The in vivo effects of leptin on circulating hormone and pituitary mRNA levels were not replicated by pair feeding ob/ob mice to match the food intake of leptin-treated mice. However, leptin did prevent the fall in hypothalamic GHRH mRNA and circulating IGF-I levels observed in pair-fed mice. These results demonstrate that leptin replacement has positive effects on multiple levels of GH axis function in ob/ob mice. PMID- 17122092 TI - Pituitary adenylate cyclase-activating peptide affects homeostatic sleep regulation in healthy young men. AB - Pituitary adenylate cyclase-activating peptide (PACAP) is involved in autonomous regulation, including timekeeping, by its action on the suprachiasmatic nucleus and on neuroendocrine secretion, energy metabolism, and transmitter release. In particular, the interactions between PACAP and the glutamatergic system are well recognized. We compared the effect of intravenously administered PACAP to that of placebo in eight healthy male subjects. PACAP in a concentration of 4x12.5 microg was administered in a pulsatile fashion hourly between 2200 and 0100. Sleep EEG was recorded from 2300 to 1000, which was also the time when subjects were allowed to sleep. Blood samples were taken every 20 min between 2200 and 0700 for the determination of cortisol, GH, and prolactin. PACAP administration led to no changes in the macro-sleep structure as assessed according to standard criteria. Spectral analysis revealed a significant reduction in the theta-frequency range in the first 4-h interval and of the spindle frequency range in the second 4-h interval of the registration period. This was accompanied by an increase in the time constant tau of the physiological delta-power decline in the course of the night, i.e., a less pronounced dynamic of the reduction of delta-power with time. This was accompanied by a trend (P<0.1) toward decreased prolactin secretion in the first 4-h period of the night. No other changes in endocrine secretion were observed. We concluded that PACAP leads to a reduction of the dynamics of homeostatic sleep regulation and prolactin secretion. Both effects are the opposite of those seen after sleep deprivation but similar to the changes after napping, i.e., a reduced sleep propensity. This implies that PACAP might be involved in homeostatic sleep regulation. PMID- 17122093 TI - Reduction of O-GlcNAc protein modification does not prevent insulin resistance in 3T3-L1 adipocytes. AB - 3T3-L1 adipocytes develop insulin-resistant glucose transport upon preincubation with high (25 mM) glucose, provided that insulin (0.6 nM) is included, Akt activation is impaired, and high glucose and insulin act synergistically. Considerable evidence suggests that increased glucose flux via the hexosamine biosynthesis pathway enhances the O-GlcNAc modification (O-GlcNAcylation) of some critical protein(s) that may contribute to insulin resistance. However, whether enhanced protein O-GlcNAcylation is necessary for the development of insulin resistance is unknown. We used two strategies to test this hypothesis. The first strategy was the overexpression of O-GlcNAcase, which removes O-GlcNAc from Ser/Thr of proteins. Cells were infected with O-GlcNAcase-expressing adenovirus (or empty virus) 5 days before they were submitted to protocols that elicit (or not) insulin resistance. O-GlcNAcase was highly expressed and functional as assessed by Western blot, O-GlcNAcase assay, and marked reduction of O GlcNAcylated proteins. The activity was mainly cytosolic. The second strategy was the expression of O-GlcNAc transferase (OGT) being markedly reduced by transfection of OGT siRNA, resulting in an approximately 90% decrease of nuclear and cytosolic OGT protein expression and similar reduction in O-GlcNAcylated proteins. Nontargeting siRNA had no effect. Preincubation in high glucose with low-dose insulin decreased the acute insulin response of glucose transport by at least 50% and impaired Akt activation. None of these parameters were affected by overexpression of O-GlcNAcase or by OGT knockout. Excess O-GlcNAcylation is one of many factors that can cause insulin resistance. It does not seem to be required for the development of glucose/insulin-induced insulin resistance of glucose transport and Akt activation in 3T3-L1 adipocytes. PMID- 17122094 TI - Defining the dosage units for colistin methanesulfonate: urgent need for international harmonization. PMID- 17122096 TI - Phototransduction, dark adaptation, and rhodopsin regeneration the proctor lecture. PMID- 17122097 TI - Demonstration of an anterior diffusional pathway for solutes in the normal human eye with high spatial resolution contrast-enhanced dynamic MR imaging. AB - PURPOSE: The present studies were conducted to determine whether a diffusional pathway for solutes exists from the ciliary body stroma to the anterior chamber of the human eye. The existence of such a pathway has been demonstrated in rabbits and monkeys, but such a pathway in humans would necessitate a shift in the physiological paradigm of the blood-aqueous barrier. METHODS: Seven normal human volunteers (five men, two women; age range, 27 to 59 years) underwent nine dynamic T1-weighted, spin-echo MR imaging studies, using intravenous, gadolinium based contrast agents. RESULTS: In all cases, signal intensity rose rapidly in the ciliary body. In all subjects, there was a measurable latent rise in signal strength (enhancement) in the anterior chamber. Signal enhancement typically occurred in the angle of the anterior chamber earlier, and to a greater degree, than within the center of the chamber. Increased signal within the posterior chamber was significantly less than in the anterior chamber, with measured increases probably attributable to volume averaging. CONCLUSIONS: These findings are consistent with the existence of an anterior diffusional pathway in the human eye. The model warrants further testing. PMID- 17122098 TI - A novel imaging technique for experimental choroidal neovascularization. AB - PURPOSE: Choroidal neovascularization (CNV) is the end point of several ocular diseases that lead to blindness. The authors developed an imaging technique for visualizing and quantifying morphologic changes associated with experimental laser-induced CNV. METHODS: CNV was induced using laser energy to disrupt Bruch's membrane. Rats were euthanatized immediately after laser injury and at 1, 2, 3, 4, 7, 14, and 60 days. Nonlasered eyes were used as the control. Eyes were enucleated and fixed, and the posterior eye cups were fluorescently labeled with markers for nuclei (DAPI; 4',6'-diamino-2-phenylindole), endothelial cells (isolectin IB4), microglia (CD11b), and filamentous actin (phalloidin). FITC dextran perfusion was compared with our technique. A confocal microscope was used to evaluate flatmounted specimens. Computer software generated three-dimensional reconstructions for qualitative and quantitative analysis of confocal image stacks. RESULTS: In nonlasered areas, RPE cells were visualized as a uniform hexagonal array. Immediately after laser exposure, a circular area devoid of fluorescent labeling was observed, indicating disruption of the choroid-Bruch's membrane-RPE complex. One day after laser exposure, cellular debris and fragmented nuclei were present, and an autofluorescent ring was visible at the site of Bruch's membrane disruption. The ring correlated with bubble formation and CNV induction. Three days after laser injury, phalloidin-labeled RPE cells and isolectin-labeled endothelial cells increased significantly, reflecting cell proliferation and migration. By day 4, isolectin-positive cells forming vascular tubes were visualized. The volume of CNV vessels increased exponentially during the next 3 days. By 7 days, a well-defined isolectin-labeled CNV network was present, and its volume was preserved for several weeks. CNV volumes calculated on the basis of FITC-dextran perfusion were significantly lower than volumes obtained using lectin-labeled samples. CONCLUSIONS: A novel imaging technique was developed that allows a three-dimensional reconstruction and measurement of laser induced CNV lesions in rat choroid/RPE flatmounts. This technique provides excellent morphologic detail and facilitates the study of critical early events in CNV, including the rupture of Bruch's membrane and the formation of endothelial clusters before vessel formation. CNV complexes are labeled at an earlier stage and more reproducibly than with FITC-dextran perfusion, providing a more accurate preclinical evaluation of antiangiogenic molecules. PMID- 17122099 TI - Retinal nerve fiber layer measurements in myopia: An optical coherence tomography study. AB - PURPOSE: To evaluate the relationship between retinal nerve fiber layer (RNFL) thickness measured by optical coherence tomography (OCT) and the axial length/refractive error of the eye. METHODS: A total of 115 eyes of 115 healthy subjects, comprising 75 eyes with high myopia (spherical equivalent [SE] < -6.0 D) and 40 eyes with low to moderate myopia (SE between -6.0 D and -0.5D), were analyzed in this cross-sectional study. Total average and mean clock hour RNFL thicknesses were measured by OCT and compared between the two myopia groups. Associations between RNFL measurements and axial length and spherical equivalent were evaluated by linear regression analysis. RESULTS: The RNFL measurements were significantly lower in the high myopia group compared with those of the low-to moderate myopia group at 12, 1, and 7 o'clock (right eye orientation). Apart from the temporal clock hours, significant correlations were evident between RNFL measurements and the axial length and spherical equivalent. The average RNFL thickness decreased with increasing axial length (r = -0.314, P = 0.001) and negative refractive power (r = 0.291, P = 0.002). A significant proportion of myopic eyes were classified as outside normal limits, with reference to the normative database. The most frequently abnormal sector was at 2 o'clock, where 16.5% of myopic eyes were outside normal limits. CONCLUSIONS: RNFL measurements vary with the axial length/refractive error of the eye. Analysis of RNFL thickness in the evaluation of glaucoma should always be interpreted with reference to the refractive status. Although the normative database provided by OCT has been helpful in identifying ocular diseases involving the RNFL, it may not be reliable in the analysis of myopic eyes. PMID- 17122100 TI - Correlation of heterogeneity for chromosome 3 copy number with cell type in choroidal melanoma of mixed-cell type. AB - PURPOSE: To study heterogeneity for chromosome 3 copy number in mixed choroidal melanoma with discrete populations of spindle and epithelioid cells using chromosome in situ hybridization (CISH) and to correlate chromosomal loss with cell type. METHODS: Twenty-two archival cases of choroidal melanoma with discrete populations of spindle and epithelioid cells were identified. CISH was used to identify chromosome 3 copy number in spindle and epithelioid areas. RESULTS: Monosomy 3 was detected in 12 (55%) of 22 choroidal melanomas. Of these, 10 (45%) had two copies of chromosome 3 in both epithelioid and spindle cells, 7 (32%) showed monosomy 3 in the epithelioid areas only, and 5 (23%) showed monosomy 3 in both epithelioid and spindle areas. CONCLUSIONS: CISH is a useful technique for analyzing chromosome copy number in different cell populations within a tumor. In mixed choroidal melanomas with discrete spindle and epithelioid cell populations, there may be heterogeneity for chromosome 3 copy number that correlates with areas of different cell type. PMID- 17122101 TI - Caspase-1 ablation protects photoreceptors in a model of autosomal dominant retinitis pigmentosa. AB - PURPOSE: Caspase-1 gene expression has been reported to be upregulated during light-induced retinal degeneration and to be reduced after neuroprotective treatments. Thus, caspase-1 may be proapoptotic in the retina. To test directly the role of caspase-1 in photoreceptor apoptosis, three mouse models were analyzed for retinal degeneration in the presence or absence of caspase-1. METHODS: Photoreceptor apoptosis was monitored in one model of induced (exposure to light) and in two models of inherited (rd1, VPP) retinal degeneration. Retinal degeneration was assessed qualitatively by light microscopy and quantitatively by the determination of free nucleosomes with ELISA or by rhodopsin measurements. Gene expression and protein levels were assessed by real-time RT-PCR and by Western blot analysis, respectively. RESULTS: Levels of caspase-1 proenzyme increased in all models of retinal degeneration concomitantly with the onset of cell death. Maturation or classic activity of caspase-1 was not detected in the retina. Ablation of caspase-1 was protective in the model of adRP (VPP mouse), but not in the two other models. Ablation of interleukin-1 receptor type 1 was without effect. Expression of monocyte chemoattractant protein (MCP)-1 increased in the model protected by caspase-1 ablation. CONCLUSIONS: Increased retinal expression of caspase-1 proenzyme may be a common marker for photoreceptor degeneration. The differential effects of caspase-1 ablation suggests a modulatory role of caspase-1 for photoreceptor apoptosis in some but not all models. Such a modulatory activity may involve a caspase-1 function different from the classic activation of interleukin-1beta. PMID- 17122102 TI - The roles of three palmitoylation sites of RPE65 in its membrane association and isomerohydrolase activity. AB - PURPOSE: RPE65, a membrane-associated protein predominantly expressed in the retinal pigment epithelium, is the isomerohydrolase in the retinoid visual cycle. Three cysteine (Cys) residues, Cys231, Cys329, and Cys330, in RPE65 have been shown to be palmitoylated and have been suggested to be responsible for its membrane association. The purpose of this study was to define the role of these Cys residues in palmitoylation, membrane association, and isomerohydrolase activity of RPE65. METHODS: The three Cys residues in RPE65 were replaced by Alanine (Ala) with site-directed mutagenesis. The mutant protein levels and subcellular localizations were determined by Western blot analysis and subcellular fractionation, respectively. Their enzymatic activities were evaluated with the in vitro isomerohydrolase activity assay. Palmitoylation of the mutants was examined by labeling of the protein with [3H]-labeled palmitic acid. RESULTS: Mutation of any single residue of these three Cys significantly reduced protein levels of RPE65. Similar to wild-type RPE65, however, all three single Cys mutants were predominantly present in the membrane fraction. Mutations of any one or two of these Cys substantially weakened the isomerohydrolase activity of RPE65, whereas mutations of all three Cys (triple mutant) completely abolished the enzymatic activity. However, this triple Cys mutant was still palmitoylated and associated with the membrane, although at a reduced level. CONCLUSIONS: There are additional yet to be identified palmitoylation sites in RPE65. The structural distortions induced by the Cys mutations may be responsible for the mislocalization and decreased isomerohydrolase activities of RPE65. PMID- 17122104 TI - Prolonged rhodopsin phosphorylation in light-induced retinal degeneration in rat models. AB - PURPOSE: The effects of various light-induced stresses on the retina were examined in the retinal degenerative rat model. METHODS: Retinal morphology and electroretinograms (ERGs) were analyzed after application of light-induced stress of several intensities (650, 1300, 2500, or 5000 lux). For evaluation of rhodopsin (Rho) function, the kinetics of Rho regeneration and dephosphorylation were studied by spectrophotometric analysis and immunofluorescence labeling with antibodies specifically directed toward the phosphorylated residues (334)Ser and (338)Ser in the C terminus of Rho. Retinal cGMP concentration was determined by ELISA. Expression levels of neurotrophic factors (FGF2, brain-derived neurotrophic factor [BDNF], platelet-derived growth factor [PDGF], and ciliary neurotrophic factor [CNTF]) were evaluated quantitatively by RT-PCR. RESULTS: Light intensity-dependent deterioration of ERG responses and thinning of the retinal outer nuclear layer were observed in wild-type and Royal College of Surgeons (RCS) rat retinas. Under dark adaptation after light-induced stress, the kinetics of Rho regeneration were not different between wild-type and RCS rat retinas. Rho dephosphorylation at (334)Ser and (338)Ser was extremely delayed in RCS rat retinas compared with wild-type without light-induced stress, but Rho dephosphorylation at those sites became slower in both RCS and wild-type rat retinas. In terms of expression of neurotrophic factors, almost no significant changes were observed between the animals after light-induced stress. CONCLUSIONS: The present study indicates that light-induced stress causes intensity-dependent deterioration in retinal function and morphology in wild-type and RCS rat retinas. Disruption of the phototransduction cascade resulting from slower kinetics of Rho dephosphorylation appears to be involved in retinal degeneration. PMID- 17122103 TI - Biological effects of thyrotropin receptor activation on human orbital preadipocytes. AB - PURPOSE: Thyrotropin receptor (TSHR) expression is upregulated in the orbits of patients with Graves ophthalmopathy (GO), most of whom have TSHR-stimulating antibodies. The authors investigated the biological effects of TSHR activation in vitro in adipose tissue, the site of orbital TSHR expression. METHODS: Activating mutant TSHR (TSHR*) or wild-type (WT) was introduced into human orbital preadipocytes using retroviral vectors. Their proliferation (Coulter counting), basal cAMP accumulation (radioimmunoassay), and spontaneous and peroxisome proliferator-activated receptor (PPARgamma)-induced adipogenesis (quantitative oil red O staining) were assessed and compared with those of nonmodified cells. QRT-PCR was used to measure transcripts of CCAT/enhancer binding protein (C/EBP)beta, PPARgamma, and lipoprotein lipase (LPL; early, intermediate, and late markers of adipogenesis) and for uncoupling protein (UCP)-1 (brown adipose tissue [BAT]). RESULTS: Expression of TSHR* significantly inhibited the proliferation of preadipocytes and produced an increase in unstimulated cAMP of 200% to 600%. Basal lipid levels were significantly increased in TSHR* (127% 275%) compared with nonmodified (100%) or WT-expressing (104%-187%) cells. This was accompanied by 2- to 10-fold increases in early-intermediate markers and UCP 1 transcripts (2- to 8-fold); LPL was at the limit of detection. In nonmodified cells, adipogenesis produced significant increases in transcripts of all markers, including LPL (approximately 30-fold). This was not the case in TSHR*-expressing cells, which also displayed 67% to 84% reductions in lipid levels. CONCLUSIONS: TSHR activation stimulates early differentiation (favoring BAT formation?) but renders preadipocytes refractory to PPARgamma-induced adipogenesis. In neither case did lipid-containing vacuoles accumulate, suggesting that terminal stages of differentiation were inhibited. PMID- 17122105 TI - Mutation causing self-aggregation in human gammaC-crystallin leading to congenital cataract. AB - PURPOSE: Many forms of congenital hereditary cataract are associated with mutations in the crystallin genes. The authors focus attention on congenital lamellar cataract, which is associated with the R168W mutation in gammaC crystallin, and congenital zonular pulverulent cataract, which is associated with a 5-bp insertion in the gammaC-crystallin gene. METHODS: To understand the molecular phenotypes-i.e., the functional defects that have occurred in the mutant gammaC-crystallin molecule in two cases described-the authors cloned, expressed, isolated, and compared the solution state structural features of these mutants with those of normal (wild-type) gammaC-crystallin. Structural models of the wild-type and mutant have been generated using comparative modeling. Circular dichroism and fluorescence spectroscopic methods were used to determine the conformation of the proteins, and temperature dependent self-aggregation was used to observe the quaternary structural features. The structural stability of the proteins was monitored with the use of chemical and thermal denaturation. RESULTS: The authors found that the 5-bp insertion led to a loss of secondary and tertiary structures of the molecule and to an enhanced tendency of self aggregation into light-scattering particles, offering a possible factor in lens opacification. The R168W mutant, on the other hand, was remarkably similar to the wild-type molecule in its conformation and structural stability, but it differed in its ability to aggregate and scatter light. CONCLUSIONS: These results support the idea that unfolding or structural destabilization is not always necessary for crystallin-associated cataractogenesis. PMID- 17122106 TI - Pax6 regulation in retinal cells by CCCTC binding factor. AB - PURPOSE: A previous study demonstrated that CTCF (CCCTC binding factor) regulates homeobox Pax6 gene expression in early embryonic stages and plays a dominant role in eye development. The purpose of the present study was to explore further the mechanism of CTCF controlling Pax6 gene expression in human retinoblastoma (Rb) cells and in the development of chicken and mouse retinas. METHODS: Northern and Western analyses were used to detect expressions of CTCF and Pax6 in Rb cells. Pax6 transcription reporter and deletion mutants were used to study the regulatory interaction between CTCF and Pax6 in Rb cells and in the retina of chicken embryos. CTCF transgenic chicken embryos and mice were established by lipofection and microinjection of linearized cytomegalovirus (CMV)-CTCF construct into fertilized eggs and mouse oocytes, respectively. Injected oocytes were implanted in the uterus of foster mothers through microinjection into the ovarian duct. The expression of CTCF and Pax6 was determined in embryo sections by immunochemistry. RESULTS: Stimulation of Rb cells with 10% FBS resulted in an increase in CTCF expression and a decrease in Pax6 expression. To study the regulatory mechanism, the Pax6 reporter and its deletion mutant activities were determined in transfected Rb cells and chicken embryonic retinas, revealing that CTCF interacts with the Pax6 gene in Rb cells through transcription control in the 5'-flanking region upstream from the Pax6 P0 promoter. Overexpression of CTCF in Rb cells suppressed Pax6 reporter activity and downregulated endogenous Pax6 expression. In contrast, downregulation of CTCF expression by knockdown of CTCF mRNA using specific small interfering (si)RNA markedly enhanced Pax6 expression in Rb cells. Further study in CTCF transgenic mouse embryos verified that overexpression of CTCF suppressed Pax6 gene expression in the retina. CONCLUSIONS: CTCF plays an important role in regulating Pax6 expression in Rb cells and in the developmental retina, and the regulation of Pax6 gene expression by CTCF in the retina is through transcriptional regulation. PMID- 17122107 TI - Dose-ranging study of lutein supplementation in persons aged 60 years or older. AB - PURPOSE: To examine the dose-response relationship between oral lutein supplementation and serum lutein concentrations in persons aged 60 years and older, with or without age-related macular degeneration (AMD). METHODS: Forty five participants with no AMD, large drusen, or advanced AMD, were randomized to receive one of three doses (2.5, 5, or 10 mg) of lutein for 6 months and to be observed for 6 additional months after the cessation of lutein supplementation. RESULTS: The mean age of the participants (33 women) was 71 years (range: 60-91). The serum lutein concentrations of each dose group were similar before supplementation, increased at 1 month, and peaked by 3 months. Median serum concentrations of the 2.5-, 5-, and 10-mg groups from baseline to month 6 increased from 18.7 to 35.1 microg/dL (2-fold increase), from 17.8 to 59.2 microg/dL (2.9-fold increase), and from 15.1 to 66.8 microg/dL (4-fold increase), respectively (all P < 0.001). The increases in lutein serum concentrations did not vary with AMD disease severity (P = 0.98). No toxicity was observed with any dose of lutein. No significant changes were detected in visual acuity or visual field tests. CONCLUSIONS: Increasing doses of lutein supplements significantly increased the serum levels of lutein and zeaxanthin, and doses up to 10 mg were safely administered. A long-term large clinical trial is necessary to investigate the safety and efficacy of lutein in reducing the risk of the development of advanced AMD. PMID- 17122108 TI - The effect of lutein and zeaxanthin supplementation on metabolites of these carotenoids in the serum of persons aged 60 or older. AB - PURPOSE: To investigate the effect of lutein supplementation at doses of 2.5, 5.0, and 10 mg/d for 6 months on distribution of these carotenoids and their metabolites in the serum of elderly human subjects, with and without age-related macular degeneration. To determine whether supplementation with lutein can interact with the serum levels of other dietary carotenoids, retinol, and alpha tocopherol. METHODS: Forty-five subjects received daily supplements of lutein (containing 5% zeaxanthin) for 6 months and were followed up for another 6 months after supplementation. Blood was collected at various intervals and lutein, zeaxanthin, and their metabolites in the sera were quantified by normal-phase high-performance liquid chromatography (HPLC)-UV/visible detection. Other dietary carotenoids, retinol, and alpha-tocopherol were identified and quantified on a C18 reversed phase HPLC column. RESULTS: After 6 months of supplementation with 10 mg of lutein, the increases in the mean serum levels from baseline were: 210 to 1000 nM/L (P < 0.0001) for lutein and 56 to 95 nM/L (P < 0.0001) for zeaxanthin. Similarly, the mean concentrations (nM/L) of carotenoid metabolites increased from 49 to 98 (P < 0.0001) for 3-hydroxy-beta,epsilon-caroten-3'-one (3'-oxolutein); 31 to 80 (P < 0.0001) for 3'-hydroxy-epsilon,epsilon-caroten-3 one; and 19 to 25 (P < 0.0001) for epsilon,epsilon-carotene-3,3'-dione. The serum levels of these carotenoids gradually decline within 6 months after supplementation. CONCLUSIONS: The increase in the serum levels of lutein/zeaxanthin correlates with increases in the serum levels of their metabolites that have previously been identified in the ocular tissues. Elderly human subjects with and without AMD can safely take supplements of lutein up to 10 mg/d for 6 months with no apparent toxicity or side effects. PMID- 17122109 TI - Expression of VSX1 in human corneal keratocytes during differentiation into myofibroblasts in response to wound healing. AB - PURPOSE: To characterize the expression of the visual system homeobox gene (VSX1) in human corneal keratocytes both in vitro and in vivo. METHODS: The expression of VSX1 was evaluated through semiquantitative RT-PCR, immunofluorescence and in situ hybridization both in corneas (either freshly obtained or wounded) and in collagenase/hyaluronidase-isolated keratocytes grown in the absence or presence of serum to promote keratocyte-to-myofibroblast differentiation. RESULTS: Quiescent or resting keratocytes normally residing in the corneal stroma or cultured in vitro in the absence of serum did not express VSX1. In wounded corneas or when cultured in the presence of serum to mimic wound-healing responses, keratocytes underwent fibroblastic transformation (with appearance of alpha-SMA and disappearance of CD-34 and keratocan signals) and started expressing VSX1. CONCLUSIONS: The results show that VSX1 is expressed in vitro and in vivo during human corneal wound healing, a process in which differentiation of corneal keratocytes into myofibroblasts occurs. These data may help to elucidate the role of VSX1 in cornea physiology suggesting a potential involvement in cornea-related diseases such as keratoconus. PMID- 17122110 TI - Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo. AB - PURPOSE: To demonstrate the application of multiphoton fluorescence (MF) and second harmonic generation (SHG) microscopy for ex vivo characterization of the structural alterations of human corneas due to keratoconus. METHODS: Buttons of keratoconic corneas derived from penetrating keratoplasty were sent for structural analysis with a custom-built multiphoton microscope. Fluorescence detected within the cytoplasm and the SHG signal obtained from collagen were used to demonstrate the morphologic changes in the corneal specimens. RESULTS: The fluorescent epithelial cells around the apical area were elongated and were aligned parallel to the adjacent collagen fibers. Parallel and centripetal distribution patterns of stromal collagen bundles were demonstrated at different depths within the keratoconic corneas. CONCLUSIONS: MF and SHG microscopy provides three-dimensional structural analysis of keratoconus ex vivo. It may provide important morphologic information for the investigation of the pathogenesis of keratoconus and may have potential in a clinical setting as an in vivo diagnostic and monitoring system for advancing keratoconus. PMID- 17122111 TI - Stimulation of collagen synthesis by insulin and proteoglycan accumulation by ascorbate in bovine keratocytes in vitro. AB - PURPOSE: Ascorbate is required for the hydroxylation of collagen that is present in the corneal stroma. The keratan sulfate proteoglycans (KSPGs) lumican and keratocan are also present, and they interact with collagen and modulate its assembly into fibrils. In this study, ascorbate was added to a defined medium containing insulin, and its effects on the synthesis of collagen and KSPGs by keratocytes were determined. METHODS: Collagenase-isolated keratocytes were cultured with or without insulin with or without ascorbate. Collagen and glycosaminoglycan synthesis was determined by collagenase digestion of incorporated 3H-glycine and by chondroitinase ABC or endo-beta-galactosidase digestion of incorporated 35SO4. KSPGs were detected by Western blot. Collagen stability was determined by pepsin digestion. Ethyl-3,4-dihydroxybenzoate (EDB) was used to inhibit collagen hydroxylation. RESULTS: Insulin stimulated the synthesis of collagen but did not affect the accumulation of lumican and keratocan. Insulin plus ascorbate, however, stimulated the synthesis of collagen and increased the accumulation of these proteoglycans. The accumulation of PGDS, a KSPG that does not interact with collagen, was not affected by ascorbate. Only the collagen synthesized in the presence of ascorbate was pepsin resistant. EDB overrode the effects of ascorbate on pepsin resistance and proteoglycan accumulation. CONCLUSIONS: The results of this study indicate that the accumulation of lumican and keratocan depends in part on the level of collagen synthesis and its hydroxylation. The interaction of lumican and keratocan with the stably folded triple helix provided by hydroxylation may also serve to stabilize these proteoglycans. PMID- 17122112 TI - Phosphatase-mediated crosstalk control of ERK and p38 MAPK signaling in corneal epithelial cells. AB - PURPOSE: To test the hypothesis that the protein phosphatases PP2A and MKP-1 are involved in controlling epidermal growth factor (EGF)-induced increases in rabbit corneal epithelial cell (RCEC) migration by mediating crosstalk between signaling pathways eliciting EGF receptor control of migration and proliferation. METHODS: Western blot analysis was used to determine the phosphorylation status of Erk1/2, p38, and the mitogen-activated protein kinase (MAPK) kinase (MEK1/2) using inhibitors of Erk1/2 or p38 and dominant-negative (d/n) Erk1 or d/n p38 cell lines. Coimmunoprecipitation was used to evaluate protein phosphatase (PP)2A and Erk1/2 interaction. Short-interfering RNA (siRNA) transfection was performed to analyze the involvement of MAPK phosphatase (MKP)-1 in crosstalk. Scratch-wound assay was used to determine EGF-dependent effects on cell migration. RESULTS: EGF (10 ng/mL) induced changes in activation of Erk1/2 and p38, which were enhanced by inhibition with 10 microM SB203580 and 10 muM PD98059, respectively. PP inhibition with sodium orthovanadate (100 microM), okadaic acid (10 nM), or Ro 31 8220 (10 microM) resulted in larger and more prolonged increases in the phosphorylation status of Erk1/2 and p38. After 1 hour, EGF induced 14-fold increases in MKP-1 protein expression. After MKP-1 siRNA transfection, EGF had induced a similar pattern of changes in the phosphorylation status in Erk1/2 and p38 following PP inhibition. EGF-induced cell migration was enhanced by Erk1/2 pathway inhibition and was accentuated after PP inhibition. Conversely, p38 pathway inhibition eliminated this response. CONCLUSIONS: EGF-induced changes in Erk1/2 and p38 phosphorylation status are dependent on PP-mediated crosstalk. This control modulates the magnitude of growth factor-induced increases in corneal epithelial cell migration. PMID- 17122113 TI - Cell surface glycoconjugate abnormalities and corneal epithelial wound healing in the pax6+/- mouse model of aniridia-related keratopathy. AB - PURPOSE: Congenital aniridia due to heterozygosity for Pax6 is associated with ocular surface disease, including keratopathy. This study investigated how defects in glycoconjugate component of the cell surface of Pax6+/- could cause the abnormal cellular migration phenotypes associated with the disease. METHODS: Immunohistochemistry, lectin-based histochemistry, conventional staining techniques, and proteomic assays were performed on eyes and cultured corneal epithelial cells from wild-type and Pax6+/- littermates. Wild-type cells were manipulated in culture to replicate the glycoconjugate abnormalities found in Pax6 heterozygotes and determine the consequences for wound healing. RESULTS: Multiple glycoconjugate defects were found in Pax6-mutant cells. Lectin cytochemistry of corneal epithelial cells suggested a partial failure of glycoprotein trafficking. Blocking cell surface carbohydrate moieties in wild type corneal cells caused wound-healing delays similar to those seen in untreated Pax6+/- cells. CONCLUSIONS: Alterations to the cell surface glycoconjugate signature of Pax6+/- corneal epithelia restrict the ability of cells to initiate migration in response to wounding. This underlies the observed wound-healing delay in cultured Pax6+/- epithelia. PMID- 17122114 TI - Mapping of a gene causing brittle cornea syndrome in Tunisian jews to 16q24. AB - PURPOSE: To map the gene that causes brittle cornea syndrome (BCS). METHODS: Five patients from four families, all of Jewish Tunisian origin, were recruited into the study. Four of the five patients had red hair. DNA from the five patients and 104 control chromosomes was typed with seven 16q polymorphic markers surrounding the hair color gene, MC1R. RESULTS: A common haplotype in the homozygous state, comprising five markers spanning 4.7 Mb on chromosome 16q24, was found in all five patients but in none of the control subjects (P < 0.00001). CONCLUSIONS: The gene that causes BCS maps to a 4.7-Mb interval, between the markers D16S3423 and D16S3425 on 16q24. PMID- 17122115 TI - Reflexive optokinetic nystagmus in younger and older observers under photopic and mesopic viewing conditions. AB - PURPOSE: To investigate the effect of age on optokinetic nystagmus (OKN) in response to stimuli designed to preferentially stimulate the M-pathway. METHOD: OKN was recorded in 10 younger (32.3 +/- 5.98 years) and 10 older (65.6 +/- 6.53) subjects with normal vision. Vertical gratings of 0.43 or 1.08 cpd drifting at 5 degrees /s or 20 degrees /s and presented at either 8% or 80% contrast were displayed on a large screen as full-field stimulation, central stimulation within a central Gaussian-blurred window of 15 degrees diameter, or peripheral stimulation outside this window. All conditions apart from the high-contrast condition were presented in a random order at two light levels, mesopic (1.8 cdm( 2)) and photopic (71.5 cdm(-2)). RESULTS: Partial-field data indicated that central stimulation, mesopic light levels, and lower temporal frequency each significantly increased slow-phase velocity (SPV). Although there was no overall difference between groups for partial-field stimulation, full-field stimulation, or low-contrast stimulation, a change in illumination revealed a significant interaction with age: there was a larger decrease in SPV going from photopic to mesopic conditions for the older group than the younger group, especially for higher temporal frequency stimulation. CONCLUSIONS: OKN becomes reflexive in conditions conducive to M-pathway stimulation, and this rOKN response is significantly diminished in older healthy adults than in younger healthy adults, indicative of decreased M-pathway sensitivity. PMID- 17122116 TI - Eye movements of patients with tunnel vision while walking. AB - PURPOSE: To determine how severe peripheral field loss (PFL) affects the dispersion of eye movements relative to the head in patients walking in real environments. This information should help to define the visual field and clearance requirements for head-mounted mobility visual aids. METHODS: Eye positions relative to the head were recorded in five patients with retinitis pigmentosa who had less than 15 degrees of visual field and in three normally sighted people, each walking in varied environments for more than 30 minutes. The eye-position recorder was made portable by modifying a head-mounted system (ISCAN, Burlington, MA). Custom data processing was implemented, to reject unreliable data. Sample standard deviations of eye position (dispersion) were compared across subject groups and environments. RESULTS: The patients with PFL exhibited narrower horizontal eye-position dispersions than did the normally sighted subjects (9.4 degrees vs. 14.2 degrees , P < 0.0001), and the vertical dispersions of patients with PFL were smaller when they were walking indoors than when walking outdoors (8.2 degrees vs. 10.3 degrees ; P = 0.048). CONCLUSIONS: When walking, the patients with PFL did not increase their scanning eye movements to compensate for missing peripheral vision information. Their horizontal scanning was actually reduced, possibly because of lack of peripheral stimulation. The results suggest that a field of view as wide as 40 degrees may be needed for closed (immersive) head-mounted mobility aids, whereas a much narrower display, perhaps as narrow as 20 degrees , may be sufficient with an open design. PMID- 17122117 TI - Leber's hereditary optic neuropathy with childhood onset. AB - PURPOSE: To characterize the clinical features of childhood-onset Leber's hereditary optic neuropathy (LHON) as defined by a pathogenic mtDNA mutation and age at onset equal to or less than 10 years of age. METHODS: Fifty-six LHON Italian pedigrees including 180 affected individuals were reviewed, and 14 of 18 patients with childhood LHON were enrolled. LHON was classified as acute bilateral, acute unilateral, slowly progressive, and subclinical, according to disease features. All patients underwent a complete ophthalmic examination and optical coherence tomography (OCT), including retinal nerve fiber layer (RNFL) and optic nerve head analysis (ONH), and were compared with age- and optic disc size-matched control groups. RESULTS: The prevalence of childhood LHON in this case series was 11.5%. Five patients had an acute bilateral course, three an acute unilateral course with subclinical signs in the fellow eye, and six a slowly progressive course. Four of five acute patients with acute bilateral disease experienced visual recovery. Slowly progressive cases presented a better visual acuity and visual field outcome than acute cases. A significant diffuse reduction of RNFL was evident in children with acute LHON compared with the control group, whereas a significant reduction of the temporal quadrant was present in the slowly progressive and subclinical LHON cases. Acute LHON children had a smaller disc area and vertical disc diameter than did the control subjects. CONCLUSIONS: This study systematically characterized for the first time the subgroup of LHON with childhood onset. The peculiar clinical and anatomic features of childhood LHON offer insights for the understanding of LHON's pathophysiology as well as a basis for the differential diagnosis of visual loss in childhood. PMID- 17122118 TI - Control of filtering bleb structure through tissue bioengineering: An animal model. AB - PURPOSE: To devise a means of providing controlled resistance between the anterior chamber and the subconjunctival space after trabeculectomy by implantation of a biodegradable, porous collagen matrix. METHODS: Matrices were implanted in the right eyes of 17 rabbits after trabeculectomy, while left eyes served as surgical controls. The scleral flap was sutured loosely, and the implant provided pressure on the scleral flap to reduce overfiltration. Trabeculectomy in the control eyes was performed with tight sutures using standard methodology. Intraocular pressure (IOP) was measured before surgery and on days 3, 7, 14, 21, and 28 after surgery. Masson trichrome and alpha-smooth muscle actin stains were used for histologic study of the filtering blebs. RESULTS: The initial postoperative IOP reduction was approximately equal, at 14% to 16%, for both groups. In the implanted group, the IOP continued to decrease to 55% below baseline at day 28 as the implant gradually degraded. In the control group, IOP had returned to the preoperative level by day 21. Histologic examination with Masson trichrome and alpha-smooth muscle actin stains showed a prominent bleb in the implanted group compared with scar formation and limited bleb formation in the control group. CONCLUSIONS: Implantation of a biodegradable, porous collagen matrix in the subconjunctival space offers the potential for a new means of avoiding early scar formation and maintaining long term IOP control by creating a loosely structured filtering bleb. PMID- 17122119 TI - A genome-wide scan maps a novel juvenile-onset primary open-angle glaucoma locus to 15q. AB - PURPOSE: To map the disease-associated locus of a family with autosomal dominant juvenile-onset primary open-angle glaucoma (JOAG). METHODS: A complete ophthalmic examination was conducted, and genomic DNA was obtained from 25 members of a Chinese family, of which eight were confirmed as having JOAG. Myocilin (MYOC), optineurin (OPTN), and WD repeat-domain 36 (WDR36) were screened for sequence alterations, by PCR and direct sequencing. Subsequently, a genome-wide scan was performed (Prism Linkage Mapping Set MD-10; Applied Biosystems, Inc., Foster City, CA). Two-point and multipoint linkage analyses were performed with the MLINK, ILINK, and LINKMAP programs. For fine mapping, additional markers flanking the most promising region on 15q were analyzed. The significance of the lod score was tested with simulation analyses by using FASTLINK. Haplotypes were constructed with Simwalk2. Three candidate genes, NR2E3, SMAD6, and CLN6, located within the critical region, were screened for mutations. RESULTS: MYOC, OPTN, and WDR36 mutations were excluded in all family members. A maximum two-point lod score of 3.31 at theta = 0.0 was obtained for the marker D15S125. Four adjacent markers, rs2030040, rs169169963, D15S153, and D15S131, gave two-point lod scores of 2.41, 2.90, 3.02, and 2.68, respectively, at theta = 0.0. Haplotype analysis and recombination mapping further confined this region to 15q22-q24 within a genetic distance of 16.6 Mb flanked by D15S1036 and rs922693. No mutations were found in the coding exons and splicing junctions of NR2E3, SMAD6, and CLN6. CONCLUSIONS: The results provide evidence for the mapping of a novel locus for JOAG at 15q22-q24. A further search for the disease-causing gene in this new JOAG locus is in progress. PMID- 17122120 TI - Measurement variability in Heidelberg Retina Tomograph imaging of neuroretinal rim area. AB - PURPOSE: To investigate the optimal frequency of imaging during follow-up to detect glaucoma progression by characterizing variability (noise) in neuroretinal rim area (RA) measured by Heidelberg Retina Tomograph (HRT; Heidelberg Engineering, Heidelberg, Germany). METHODS: RA noise was estimated from patient data and characterized by fitting theoretical distributions to the observed data. Multilevel regression was used to determine factors that significantly affect noise. Computer simulations of disease progression were performed by adding noise generated from the distribution derived from the observed data to the average rate of loss in RA estimated from longitudinal data. Rates of detection of disease progression were investigated for various progression rates, follow-up periods, and rates of imaging. RESULTS: Noise was not normally distributed and was best characterized by the hyperbolic distribution, which fit averages well while allowing for extreme values. Noise was greatly influenced by image quality, but age did not have a significant effect. Rates of detection improved for more frequent imaging, better quality images, and faster rates of disease progression. CONCLUSIONS: Noise in HRT measurement of RA is well characterized by the hyperbolic distribution. Sensitivity of detection improves with more frequent testing, but if consistently poor-quality images are yielded for a patient, the probability of detection is low. Results from this work could be used to tailor individual follow-up patterns for patients with different rates of RA loss and image quality, especially in a clinical trial setting. PMID- 17122121 TI - A comparison between multifocal and conventional VEP latency changes secondary to glaucomatous damage. AB - PURPOSE: To compare latencies of conventional visual evoked potentials (cVEPs) and multifocal VEPs (mfVEPs) in the same patients. Previous reports of prolonged cVEP latency suggest a vehicle for detecting abnormal ganglion cells and for monitoring neuroprotection. METHODS: Seventy-five glaucomatous eyes (47 patients), 75 eyes with suspected glaucoma (46 patients), and 41 control eyes (22 subjects) underwent achromatic automated perimetry and mfVEP and cVEP testing. The mfVEP stimulus was a scaled dart board with 60 sectors; each sector was a pattern-reversing checkerboard. The cVEP stimulus was a reversing checkerboard with checks of either 15 minutes or 60 minutes in width. RESULTS: Relatively few glaucomatous eyes had latencies that fell outside the range of control eyes, and there was little difference between the cVEP and mfVEP results. In the glaucoma group, 12.3% (15 minutes cVEP), 8% (60 minutes cVEP), and 17.3% (mfVEP) of the eyes and 5.3% (15 minutes cVEP), 6.7% (60 minutes cVEP), and 5.3% (mfVEP) of the suspect eyes exceeded the normal range. The glaucomatous eyes had, on average, relatively small increases in latency, compared with the control or suspect groups. Further, the latency of both the mfVEP and cVEP bore no obvious relationship to the mean deviation of the visual field. CONCLUSIONS: Contrary to previous reports, prolonged VEP delays were present in a minority of patients with glaucoma. Either a delayed VEP is not a good indicator of damaged, as opposed to dead, retinal ganglion cells, or there are relatively few patients who exhibit evidence of damaged ganglion cells. PMID- 17122122 TI - Corneal thickness- and age-related biomechanical properties of the cornea measured with the ocular response analyzer. AB - PURPOSE: The Ocular Response Analyzer (ORA) is a new instrument that measures the corneal biomechanical response (corneal hysteresis, CH) to rapid indentation by an air jet. CH is the difference in applanation pressures (P1, P2) between the rising and falling phases of the air jet. The investigation had two parts: a characterization study and a validation study. In the characterization study, the purposes were to investigate the intraocular pressure (IOP)-dependence of CH and to characterize the performance of the ORA. In the validation study, the purposes were to investigate the association between CH and both age and central corneal thickness (CCT) and the agreement between ORA and Goldmann applanation tonometer (GAT) IOP measurements. METHODS: For the characterization study, data were collected from 105 untreated subjects (45 ocular hypertensive patients and 60 normal subjects; mean age, 60 years, range, 26-82). GAT and ORA measurements were performed before and after IOP lowering of one randomly selected eye with apraclonidine drops. The change in P1 and P2 (arbitrary units) in relation to change in GAT IOP was analyzed to calibrate the instrument. The relation between P1, P2, and CCT was explored and ORA IOP was derived from the analyses. For the validation study, ORA and GAT IOP and CCT were measured in 144 eyes of 144 untreated subjects (mean age, 58 years; range, 19-83). The characterization calculations were applied to the dataset and values of CH and ORA IOP were calculated. The relationship between CH and both subject age and CCT was determined. The associations between CH and CCT and between ORA and GAT IOPs, were investigated by linear regression analysis. The agreement between measuring devices was calculated. RESULTS: In the characterization study, P1 changed by 6.41 arbitrary units for every 1-mm Hg change in GAT IOP. CH (P1 - P2) changed by -1.60 arbitrary units for every 1-mm Hg change in GAT IOP. For each unit change in P2, P1 changed by 1.27 units. From this association a new IOP-independent corneal factor was derived [P1 - (P2/1.27)] and is termed the corneal constant factor (CCF; mm Hg). ORA IOP normalized for CCF was defined as P2 - CCF (mm Hg). The CCF (mm Hg) was associated with CCT (micrometers) and with age: CCF = [(0.036 . CCT) - (0.028 . age)] + 1.06 (adjusted r2 = 0.34; P < 0.0001 for CCT, P = 0.007 for age). Normalized ORA IOP measurements were not associated with CCT. GAT IOP was associated with CCT and CCF-more strongly with the latter: GAT IOP = (0.03 . CCT)+1.52 (r2 = 0.06, P = 0.002); GAT IOP = (0.65 . CCF) + 4.5 (r2 = 0.13, P < 0.0001). The mean difference (95% limits of agreement) between GAT and normalized ORA IOP was 0.1 (-6.6 to +6.8) mm Hg. CONCLUSIONS: The CCF describes an IOP independent biomechanical property of the cornea that increases with thicker CCT and decreases with greater age. It is moderately strongly associated with CCT and yet explains more of the interindividual variation in GAT IOP than does CCT. Normalized ORA IOP measurements are not associated with CCT. PMID- 17122123 TI - Automated analysis of heidelberg retina tomograph optic disc images by glaucoma probability score. AB - PURPOSE: To compare the diagnostic performance of the Heidelberg Retinal Tomograph's (HRT; Heidelberg Engineering GmbH, Dossenheim, Germany) glaucoma probability score (GPS), an automated, contour line-independent method of optic disc analysis with that of the Moorfields regression analysis (MRA). METHODS: HRT images were obtained from one eye of 121 patients with glaucoma (median age, 70.2 years; median mean deviation [MD], -3.6 dB, range, +2.0 to -9.9 dB) and 95 healthy control subjects (median age, 59.7 years; median MD -0.1 dB, range +2.5 to -3.7). The diagnostic performances of GPS and MRA were evaluated by including borderline classifications, either as test negatives (most specific criteria) or as test positives (least specific criteria). Agreement between global and sectoral data of both analyses was established. Logistic regression analyses were performed to evaluate the effect of covariates such as optic disc size and age on the classification outcomes of both the GPS and the MRA. RESULTS: In 8 (7%) patients with glaucoma and 10 (11%) control subjects, the GPS failed to provide a complete global and sectoral optic disc classification. Although we could not identify a single distinct cause of this failure in the glaucoma group, failures in the control subjects occurred most often (7/10) with small and crowded optic discs. In subjects who were successfully classified at least globally by the GPS (117 patients with glaucoma, 88 control subjects), the diagnostic performances of GPS and MRA were similar (areas under the receiver operating characteristic [ROC] curve of 0.78 and 0.77, respectively; P > 0.1). With the GPS, sensitivity and specificity were 59% and 91% (most specific criteria) and 78% and 63% (least specific criteria), respectively. Combining GPS and MRA did not increase diagnostic performance significantly (ROC area of combined classifiers, 0.81). Both GPS and MRA were affected by disc size. In patients with glaucoma as well as healthy control subjects, the odds of a positive GPS classification (borderline or outside normal limits) increased by 21% (95% confidence interval [CI], 12% 30%) for each 0.1 mm2 increase in optic disc area. With the MRA, the corresponding increase was 15% (95% CI, 7%-23%). Optic disc area alone accounted for approximately 30% and 22% of the explained variance with the GPS and MRA, respectively (P < 0.001). The proportional-odds logistic regression confirmed that optic disc size affected mainly the tradeoff between true- and false positive classifications (criterion) rather than the absolute performance of the analyses (area under the ROC curve). There was some evidence of an age effect with the MRA, which showed a 53% (95% CI, 16%-102%) increase in the odds of a positive test (borderline or outside normal limits) associated with each decade of age (P = 0.002), but no age effects were observed with the GPS (P > 0.1). CONCLUSIONS: The diagnostic performance of the contour line-independent GPS analysis is similar to that of the MRA. However, clinicians should be aware of the strong size dependence of both GPS and MRA. In large optic discs, both GPS and MRA are likely to produce many false-positive classifications. Correspondingly, the sensitivity to early damage is likely to be low in small optic discs. There is a need for automated classification systems that explicitly address the size dependence of current analyses. PMID- 17122124 TI - Structure and function in glaucoma: The relationship between a functional visual field map and an anatomic retinal map. AB - PURPOSE: To examine the relationship between an anatomic map relating the retinal nerve fiber layer (RNFL) distribution to the optic nerve head and a functional map derived from the interpoint correlation of raw sensitivities in visual field (VF) testing. METHODS: Previously, interpoint correlations were generated for all possible pairs of VF test points in a dataset of 98,821 Humphrey VF test results taken from the Moorfields Eye Hospital archive. The relationship between these correlations and the physical distance between the VF test point pairs was evaluated by Pearson's correlation coefficient and multiple regression analysis. The distance between the pairs of VF test points was calculated in two ways. First, the anatomic map was used to estimate the angular distance at the optic nerve head (ONH), between the RNFL bundles corresponding to the VF test points in each pair (ONHd). Second, the retinal distance between pairs of test points was calculated from the Humphrey VF template (RETd). A best-fit model for predicting functional correlation (FC) from ONHd and RETd was constructed and used to formulate a filter incorporating the anatomic-functional correlation data. RESULTS: All scatterplots showed a negative association between interpoint retinal sensitivity correlation values and distance between points: ONHd (R2 = 0.60) and RETd (R2 = 0.33). The raw sensitivity correlation values could be predicted from a multiple regression model using ONHd, RETd, and a combined interaction of ONHd and RETd (R2 = 0.75, P < 0.00001). The construction of a new filter was based on the equation FC = 0.9325 - (0.0029 . ONHd) - (0.0077 . RETd) + (0.0001 . ONHd . RETd). CONCLUSIONS: A good level of association was observed between the strength of correlation between points in the VF and the relative location of those test points in the peripheral retina and in corresponding RNFL bundles at the ONH. These results help to validate the relationship between structure and function and may be of use in the further refinement of physiologically derived VF filters to reduce measurement noise. PMID- 17122125 TI - Microtubule contribution to the reflectance of the retinal nerve fiber layer. AB - PURPOSE: The reflectance of the retinal nerve fiber layer (RNFL) arises from light scattering by cylindrical structures oriented parallel to ganglion cell axons. In amphibian retinas, at 440 nm, microtubules (MTs) contribute about one half of RNFL reflectance. In rodent retinas, MTs are the only structure contributing to RNFL birefringence. To increase understanding of the anatomic basis for clinical RNFL measurements, this study was conducted to evaluate the MT contribution to RNFL reflectance in rodent retinas by using the MT depolymerizing agent colchicine. METHODS: Reflectance of nerve fiber bundles in isolated rat retinas was measured at 460, 580, and 830 nm with a multispectral imaging reflectometer. Images were taken frequently over an extended period. During baseline, the tissue was perfused with a physiological solution. During a treatment period, the solution was switched either to a control solution or to a solution containing colchicine. RESULTS: Because of the high reflectance of the RNFL, nerve fiber bundles appeared as bright stripes against a darker retina. The reflectance of bundles was relatively stable in control experiments. With colchicine treatment, however, bundle reflectance at first decreased rapidly and then became stable. After 70 minutes of colchicine treatment, RNFL reflectance had declined to approximately 50% below baseline at all wavelengths. CONCLUSIONS: MTs contribute to RNFL reflectance at all wavelengths. Unlike RNFL birefringence, however, which totally disappears after colchicine treatment, about one half of RNFL reflectance remained after colchicine treatment. This result suggests that, in addition to MTs, other mechanisms may contribute to RNFL reflectance. PMID- 17122126 TI - SNPs and interaction analyses of noelin 2, myocilin, and optineurin genes in Japanese patients with open-angle glaucoma. AB - PURPOSE: To evaluate the noelin 2 gene as a disease-causing factor for open-angle glaucoma (OAG) and the interactions between the noelin 2 (OLFM2), optineurin (OPTN), and myocilin (MYOC) genes. METHODS: OLFM2 was analyzed in 770 Japanese subjects including 215 patients with elevated intraocular pressure (IOP), 277 with normal IOP, 38 with juvenile open-angle glaucoma, and 240 control subjects. Two single-nucleotide polymorphisms (SNPs) in OPTN (c.412G-->A and c.603T-->A) and one SNP in MYOC (c.227G-->A) were examined. Single genes were investigated by univariate analysis and the gene-gene interactions by logistic regression analysis. Associations between genotypes and clinical characteristics at the time of diagnosis were examined. RESULTS: In OLFM2, 12 sequence variants were identified in 770 Japanese subjects. Arg144Gln (exon 4) was identified in two (0.3%) of the patients and in none of the control subjects. Combinations of OLFM2/317A and OPTN/412A or OLFM2/1281T and OPTN/412A were associated with patients with elevated IOP (P = 0.018 or P = 0.012, respectively). The combination of OLFM2/317G and OPTN/603A was significantly associated with elevated IOP (P = 0.018). No significant association was detected between SNPs in OLFM2 and in MYOC. Patients with normal IOP and with OLFM2/678A+OPTN/412G or OLFM2/1281C+OPTN/412G had significantly worse visual field scores (P = 0.022 or 0.030, respectively). CONCLUSIONS: The Arg144Gln mutation in OLFM2 is a possible disease-causing mutation in Japanese patients with OAG. Common polymorphisms in OLFM2 and OPTN may interactively contribute to the development of OAG, indicating a polygenic etiology. PMID- 17122127 TI - B7+ iris pigment epithelial cells convert T cells into CTLA-4+, B7-expressing CD8+ regulatory T cells. AB - PURPOSE: To determine whether iris PE (IPE) promotes the generation of regulatory T-cells (Tregs) with cell contact via B7-2/CTLA-4 interactions. METHODS: T cells were cocultured with IPE cells obtained from eyes of normal and B7-deficient mice, x-irradiated, and used as regulators. IPE T regulator cells (IPE Tregs) of normal and CD28- or CTLA-4-deficient mice were established. Target bystander T cells were established from normal splenic T cells with anti-CD3 antibodies. T cell activation was assessed for proliferation by [3H]-thymidine incorporation. Neutralizing anti-B7-1 and/or B7-2 antibodies, anti-CTLA-4 antibodies, CTLA-4-Ig fusion proteins were used to abolish regulatory function. IPE-exposed CD8+ T cells were evaluated for expression of B7, CTLA-4, and Foxp3 by using RT-PCR and flow cytometry. CD8+ IPE Tregs were depleted of B7-2+ and CTLA-4+ T cells and assayed for suppressive activity by adding them to bystander T cells. RESULTS: T cells acquired T regulatory activity when exposed to cultured IPE. Ciliary body PE cells did not promote conversion of T cells into Tregs. IPE converted CD8+, but not CD4+, T cells into Tregs by direct cell contact. In the conversion, IPE and responding T cells must both express endogenously synthesized B7-1 and B7-2, and the T cells must also express CTLA-4. Expression of CD28 molecules was not necessary for Treg generation. In addition, the CD8+ Tregs that fully suppress activation of bystander T cells expressed Foxp3. CONCLUSIONS: IPE cells promote conversion of T cells into Tregs solely through a contact-dependent mechanism. T cells exposed to IPE cells acquire full regulatory capacity. PMID- 17122128 TI - RPE cells resist bystander killing by CTLs, but are highly susceptible to antigen dependent CTL killing. AB - PURPOSE: Retinal pigmented epithelial (RPE) cells maintain the blood-retinal barrier, sustain retinal photoreceptor cell health and function, and may play a role in ocular immune privilege. If RPE immunomodulatory activities were antigen specific, their expression would require antigen presentation. In a study of antigen processing and major histocompatibility complex (MHC) class I-restricted presentation by RPE cells, the cells' sensitivity to the activity of cytotoxic T lymphocytes (CTLs) was determined. METHODS: RPE was cultured, with and without proinflammatory cytokines and antigen, followed by the addition of beta galactosidase (beta-gal)-specific CTLs. Cytotoxic activity was measured by the CTL-dependent activation of caspase-3 in the RPE. Sensitivity to the CTLs was used to evaluate the activity of pathways of antigen processing and presentation using an antigen (beta-gal) that was either applied to or expressed in RPE. RESULTS: RPE cells were sensitive targets for activated CTL-mediated killing in vitro only if prepulsed with cognate peptide, or if beta-gal-expressing RPE was pretreated to induce upregulation of immunoproteasome. Activated CTLs induced apoptosis in RPE within 3 hours of coculture with antigen-positive RPE monolayers. Application of CTLs in a resting state to antigen-positive RPE led to their activation in the absence of exogenous antigen-presenting cells (APCs). This antigen-dependent activation and killing required 24 hours of co-incubation of RPE with resting CD8 T cells specific for beta-gal. Although RPE cells are highly phagocytic, functional evidence for processing that allowed phagocytosed antigens to load into class I MHC was not detected. RPE was minimally sensitive to bystander killing by activated CTLs. CONCLUSIONS: Although there are many reports of T-cell inhibition by RPE, we found that CTLs efficiently killed RPE cells by induction of apoptosis in an antigen-dependent manner. The survival of RPE in the face of extensive CTL destruction of adjacent photoreceptor cells in vivo appears to be based on their insensitivity to injury via bystander mechanisms. PMID- 17122129 TI - Inhibition of aldose reductase prevents lipopolysaccharide-induced inflammatory response in human lens epithelial cells. AB - PURPOSE: Bacterial infections are one of the major causes of human eye disease. Because the bacterial endotoxin lipopolysaccharide (LPS) is known to cause cytotoxicity through oxidative stress and an earlier study has shown that aldose reductase (AR) mediates oxidative stress signals, the purpose of this study was to investigate the anti-inflammatory effects of AR inhibition on LPS-induced activation of NF-kappaB-dependent signals in human lens epithelial cells (HLECs). METHODS: Growth-arrested HLECs were cultured without or with AR inhibitors or transfected with an AR small interfering (si)RNA. Subsequently, the cells were stimulated with LPS (1-10 mug/mL) for 24 hours. The cell viability was assessed by cell counts and MTT assay, and apoptosis was measured by nucleosomal degradation. Electrophoretic mobility gel shift assays were performed to determine the activation of NF-kappaB and AP1. The levels of nitric oxide, MMP-2, MMP-9, Cox-2, and TNF-alpha were measured by using specific ELISA kits. Western blot analysis was performed to determine the cleavage of poly(ADP-ribose) polymerase (PARP) and the activation of PKC and mitogen-activated protein kinase (MAPK). RESULTS: Bacterial LPS caused apoptosis of HLECs. Inhibition of AR by two structurally unrelated inhibitors, sorbinil and tolrestat, or ablation by AR siRNA prevented the LPS-induced apoptosis, activation of caspase-3 and cleavage of PARP protein. Inhibition of AR in HLECs also prevented the LPS-induced activation of redox-sensitive transcription factors such as NF-kappaB and AP1 and their downstream signals that lead to expression of Cox-2, MMP-2, MMP-9, and TNF alpha proteins. In addition, inhibition of AR prevented LPS-induced activation of protein kinases upstream to NF-kappaB activation such as PKC and MAPK in HLECs. CONCLUSIONS: The results indicate that AR mediates the bacterial endotoxin signaling that could damage HLECs by regulating the signals that activate the redox-sensitive transcription factor NF-kappaB and cause inflammation. Thus, inhibition of AR could be a therapeutic target for Gram-negative bacterial infection-induced visual complications. PMID- 17122130 TI - Biological response in various compartments of the rat lens after in vivo exposure to UVR-B analyzed by HR-MAS 1H NMR spectroscopy. AB - PURPOSE: The purpose of the present study was to investigate metabolic changes in different compartments of the rat lens (anterior, nuclear, posterior, and equatorial) after exposure to an acute double threshold dose of ultraviolet-B radiation (UVR-B) by using high-resolution magic angle spinning (HR-MAS) (1)H nuclear magnetic resonance (NMR) spectroscopy and pattern recognition (PR) METHODS: methods. One eye in each of 28 6-week-old female albino Sprague-Dawley rats was exposed to in vivo 7.5 kJ/m2 UVR-B for 15 minutes. The contralateral eye was left unexposed. One week after irradiation, all rats were killed, and both lenses were isolated. Each lens was cored by a trephine, and the cylinder was sliced into three portions (anterior, nuclear, and posterior). The lens material that remained after the coring process was analyzed as the equatorial region. Analysis of lens metabolism was performed by HR-MAS 1H NMR spectroscopy (14.1 T; Avance DRX600; Bruker BioSpin GmbH, Rheinstetten, Germany), and the metabolic profiles were statistically analyzed by the PR method of principal component analysis (PCA). RESULTS: Metabolic differences were detected among the compartments in the lens, both in samples from the contralateral nonexposed lenses and in samples from lenses exposed to in vivo UVR-B. In the rat lens, exposure to UVR-B caused changes in GSH, phosphocholine, myo-inositol, succinate, formate, and adenosine triphosphate (ATP)/adenosine diphosphate (ADP) and in levels of the amino acids phenylalanine, taurine, hypo-taurine, tyrosine, alanine, valine, isoleucine, and glutamate, that varied among lens compartments. CONCLUSIONS: HR-MAS 1H NMR spectroscopy, combined with PR methods (PCA), is effective for analysis of separate parts of the intact rat lens. To understand the biochemistry of the lens, it is important to divide the lens into sections, representing functionally and anatomically distinct compartments. PMID- 17122131 TI - Contrast acuity in cataracts of different morphology and association to self reported visual function. AB - PURPOSE: To evaluate the relationship between contrast acuity at declining contrast levels and the type and density of lens opacity in cataract. METHODS: Contrast acuity at declining contrast levels was determined with the Holladay Contrast Acuity Test, in relation to the type and density of age-related cataract in 180 patients with bilateral cataract and 20 control subjects with normal macular function. Cataracts were graded according to the Lens Opacities Classification System (LOCS) III of nuclear color (NC), nuclear opalescence (NO), cortical (C), and posterior subcapsular (P) cataract. Best-corrected visual acuity and near contrast acuity were determined in randomized order monocularly in both eyes. Visual difficulties in everyday life were evaluated, using the VF 14 questionnaire and the Cataract Symptom Score. RESULTS: The contrast-dependent effect of cataract on contrast acuity was statistically significant (P < 0.001; two-way ANOVA). In the comparison of early, intermediate, and advanced nuclear, nuclear-cortical, and posterior subcapsular cataracts (PSCs), significantly reduced contrast acuity scores were found for the PSC groups (P < 0.001). Comparison of nuclear and nuclear-cortical cataracts showed the contrast acuity scores to be comparable at all contrast levels (P > 0.05). High correlation coefficients were found between the LOCS III P score and the contrast acuity measurements (r = 0.77-0.84; P < 0.001). In contrast, the correlation coefficients of the NO, NC, and C scores were considerably lower (r = 0.45-0.66; P < 0.001). High correlation coefficients were also found between the contrast acuity measurements and self-reported functional vision. CONCLUSIONS: The statistically significant, contrast-dependent effect of cataract on contrast acuity supports the clinical relevance of recording visual acuity at low contrast levels in patients with age-related cataract. Particularly, the severity of PSC has a strong influence on the impairment of contrast acuity. Contrast acuity corresponded closely to the self-reported visual difficulties in everyday life. PMID- 17122132 TI - RGC sensitivity to mild systemic hypoxia. AB - PURPOSE: Systemic hyperoxia, hypercapnia, and hypoxia are known to alter retinal perfusion. The effects such experimentally induced systemic blood gas perturbations have on inner neuroretinal function in humans were examined. METHODS: Twenty healthy adults participated in each of three test sessions. The effects of breathing pure oxygen (O2), carbogen, or a hypoxic gas on the pattern electroretinogram (pERG) were investigated. The stimulus consisted of high contrast, black-and-white, 48-min arc checks reversing at 1 Hz. pERGs were recorded with a Dawson-Trick-Litzkow electrode at the end of 5 minutes of breathing room air, 5 minutes of breathing the test gas, immediately after the flow of gas was stopped, and 10 minutes after the flow of gas was stopped. RESULTS: Amplitudes and implicit times of the major positive (P50) and negative (N95) components of the pERG were not altered during the pure O2 and carbogen breathing sessions. Although the amplitude and implicit time of P50 were not modified significantly with systemic hypoxia, they were depressed and delayed, respectively, for N95. CONCLUSIONS: Inner neuroretinal function remained unchanged during increased blood O2 and carbon dioxide levels known to alter retinal blood flow, but it was altered during decreased blood O2 levels. Overall, these results indicate that the generators of P50 are resistant to systemic hyperoxia, hypercapnia/hyperoxia, and hypoxia. They further indicate that the generators of N95, namely the retinal ganglion cells, are particularly sensitive to transient, mild systemic hypoxia. PMID- 17122133 TI - Chloride secretion by porcine ciliary epithelium: New insight into species similarities and differences in aqueous humor formation. AB - PURPOSE: To investigate the electrophysiology and mechanisms of chloride (Cl-) transport across the ciliary body-epithelium (CBE) of the porcine eye. The pig is widely believed to be a good model for studying human physiology. Current results strengthen our understanding of the physiology of aqueous humor formation (AHF). METHODS: Freshly isolated porcine CBE were maintained in modified Ussing-Zerahn type chambers. The effects of the bathing anion substitution (Cl- and HCO3-) and transport inhibitors including bumetanide, 4,4'-diisothiocyanatostilbene-2,2' disulfonic acid disodium salt (DIDS), heptanol, and two chloride channel inhibitors, 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB), and niflumic acid, on the electrical properties and transepithelial Cl- transport were investigated. RESULTS: Viable porcine CBE preparations were maintained in vitro. A spontaneous transepithelial potential difference (PD) of approximately 1 mV was found across the CBE (aqueous side negative). The magnitudes of the PD and short circuit current (I(sc)) were found to be dependent on both the bathing Cl- and HCO3- concentrations. In short-circuited conditions, a significant net Cl- transport (1.01 microEq x h(-1) x cm(-2); n = 109; P < 0.001) in the stromal-to aqueous direction (J(net)Cl) was detected. The magnitude of the Cl- current carried by the J(net)Cl was approximately 2.2 times the measured I(sc), suggesting there was cation (e.g., Na+) transport along with Cl- and/or anion transport (e.g., HCO3-) in the opposite direction. Bilateral bumetanide (0.1 mM) reduced the J(net)Cl by approximately 56% while stromal DIDS (0.1 mM) produced no inhibition. Instead, aqueous DIDS (0.1 mM) triggered a sustained stimulation of both I(sc) and J(net)Cl. Even if bilateral DIDS was used at a higher concentration (1 mM), together with bilateral dimethylamiloride (DMA, 0.1 mM), no inhibition of the I(sc) was observed. Bilateral heptanol (3.5 mM) drastically reduced the I(sc) and J(net)Cl. NPPB (0.1 mM), a common chloride channel inhibitor, did not inhibit the J(net)Cl, whereas NFA (1.0 mM) virtually abolished it. CONCLUSIONS: In the porcine eye, active secretion of Cl- into aqueous was identified that may act as a driving force for AHF. The bumetanide-sensitive Na+/K+/2Cl- cotransporter (NKCC) clearly contributes to the Cl- uptake into the pigmented epithelium (PE), whereas the DIDS-sensitive Cl-/HCO3- anion exchanger (AE) may exert a minor role. The intercellular gap junctions couple the porcine ciliary bilayers and thus the transepithelial Cl- transport, as in other species. The Cl- channel/efflux pathway located in the nonpigmented epithelium (NPE) is niflumic acid-sensitive but NPPB-insensitive. We also hypothesize that the AE located on the NPE may regulate the activity of a putative Cl- channel on the basolateral membrane facing aqueous via modulation of the intracellular pH (pHi). This work reinforces the general consensus that active secretion of Cl- is the major driving force of AHF in mammalian eye and further substantiates the existence of species differences in the mechanism that accomplishes transepithelial Cl- transport. PMID- 17122134 TI - Evaluation of the new photosensitizer Tookad (WST09) for photodynamic vessel occlusion of the choroidal tissue in rabbits. AB - PURPOSE: To determine the efficacy of Tookad (WST09; Negma-Lerads, Magny-Les Hameaux, France) photodynamic therapy (T-PDT) by evaluating the angiographic and histologic closure of choroidal vessels at different radiance exposures, drug dosages, and intervals between photosensitizer injection and laser application in a rabbit model. METHODS: Chinchilla Bastard rabbits were injected intravenously with three different dye concentrations (2.5, 5, and 10 mg/kg) before application of light. In every group T-PDT was performed at four different times after injection: 5, 15, 30, and 60 minutes with different radiance exposures ranging from 200 to 3 J/cm2. Fundus photographs and fluorescein angiograms were obtained 90 minutes after injection. Follow-up angiographies were performed at days 1, 3, 7, and 14 after initial treatment. Histology was performed in selected cases immediately after treatment and on days 1, 3, and 7. RESULTS: Immediately after irradiation, most of the visible lesions were angiographically hyperfluorescent due to damaged vessel endothelium and associated RPE damage. Lesions from high radiance exposures revealed immediate hypofluorescence, indicating vessel closure. Hypofluorescent lesions appeared mainly during day 1 (all lesions angiographically visible, some hypofluorescent) to day 3 (all lesions hypofluorescent) after treatment. At day 7, ophthalmoscopically visible hyperpigmentation took place in all lesions. ED50 thresholds for angiographic hypofluorescence determined at day 3 after treatment with 2.5 mg/kg were 18.8 J/cm2 (5 minutes), 62.0 J/cm2 (15 minutes), and >100 J/cm2 (30 minutes); with 5 mg/kg, 8.4 J/cm2 (5 minutes), 22.8 J/cm2 (15 minutes), 54.5 J/cm2 (30 minutes), and >100 J/cm2 (60 minutes); and with 10 mg/kg, 11.7 J/cm2 (30 minutes) and 54.1 J/cm2 (60 minutes). Histology of the angiographically hypofluorescent lesions revealed vessel thrombosis in all groups 1 hour after PDT up to 7 days after treatment. Sparing of photoreceptors indicated selectivity of T-PDT; however, slight damage was partly observable. After 7 days, localized proliferation of the RPE cells was noted and was enhanced 14 days after treatment. CONCLUSIONS: T-PDT has the potential to achieve selective choroidal vessel occlusion with proper parameter selection, such as (1) 2.5 mg/kg, 5 minutes, 100 J/cm2; (2) 5 mg/kg, 5 minutes, 25 J/cm2; or (3) 5 mg/kg, 15 minutes, 50 J/cm2; however, slight damage to the photoreceptors cannot be ruled out. RPE proliferation indicates primary RPE damage due to PDT, also described with the use of all other photosensitizers. PMID- 17122135 TI - Development of the electroretinographic oscillatory potentials in normal and ROP rats. AB - PURPOSE: To study the development of the electroretinographic (ERG) oscillatory potentials (OPs) in rats and to compare normal OPs with those in a rat model of retinopathy of prematurity (ROP). METHODS: Following a longitudinal design, ERG responses to a greater than 5 log unit range of full-field stimuli were recorded in dark-adapted rats at postnatal day (P) 18, P31, P47, and P67. The ERG records were digitally filtered (60-235 Hz), and the trough-to-peak amplitudes and implicit times of OP2, OP3, OP4, and OP5 were measured. Additionally, rats with oxygen-induced retinopathy, a model of ROP, were studied at P31. RESULTS: Generally, OP amplitude increased and implicit time decreased with increasing stimulus intensity. The shape of the stimulus-response functions changed with age. The amplitudes of OP2, OP3, and OP4 were largest at P31. OP5 was largest at P47. All OPs were significantly affected in ROP rats; OP5 was least affected by ROP. CONCLUSIONS: A prolonged normal course of OP development, which featured waxing and waning of amplitudes, was observed and might have been consequent to maturation and then to final refinements of inner retinal circuitry. In ROP rats, marked attenuation of early OPs was consistent with persistent dysfunction of photoreceptors, and significant attenuation of the late OP5 was evidence of compromised function of inner retinal circuitry. PMID- 17122136 TI - Expanded genome scan in extended families with age-related macular degeneration. AB - PURPOSE: To investigate further the genetic contribution to age-related macular degeneration (AMD), increasing the power of a previous analysis and reproducing the original findings. METHODS: A large cohort of families with this condition was assembled, and an expanded genome scan was performed with 556 microsatellite markers. In 2003, the results were reported of a genome-wide linkage analysis of 70 of these pedigrees. Members of 51 new families have now been ascertained and many of the original pedigrees expanded. Parametric and nonparametric linkage analyses were performed with a denser map of markers. In addition, analyses were performed with the sample stratified by age at ascertainment and by two major advanced phenotypes for the disease: neovascular AMD (choroidal neovascularization) and geographic atrophy. RESULTS: The results corroborate the macular degeneration-susceptibility loci consistently reported by the authors and others in genome-wide scans. New loci were identified, including the finding of a two-point HLOD of 3.70 at 6q25.2. CONCLUSIONS: The results suggest that the use of families enriched in predisposition to AMD has legitimacy. Genetic analyses of a genome-wide scan performed on our large cohort of families add further confirmatory evidence that susceptibility loci lie on 1q, 3p, 9q, and 10q. Furthermore, new loci have been identified, including a locus on 6q. PMID- 17122137 TI - Dynamic and quantitative analysis of choroidal neovascularization by fluorescein angiography. AB - PURPOSE: In this study, the authors sought to develop and characterize techniques for measuring changes in choroidal neovascularization (CNV) lesion size and fluorescence over time for quantitative analysis of fluorescein angiograms. METHODS: Initial assessment of the quantitative technique was made by retrospectively analyzing digital fluorescein angiograms taken before and 3 months after photodynamic therapy (PDT) for CNV (6 patients, group 1). The method was then applied prospectively to digital fluorescein angiograms (baseline and day 71) obtained on 12 patients taking part in a clinical trial investigating the effect of vascular endothelial growth factor (VEGF) Trap in CNV (group 2). Two masked observers, with the use of image processing, measured the area of hyperfluorescence and fluorescence intensity above background. Values for each image were plotted against time after dye injection to generate curves, and each area under the curve (AUC) was calculated. RESULTS: The physician who treated the patients in group 1 judged the condition of three patients to be improved and of three to be worse 3 months after PDT. Masked retrospective grading of fluorescein angiograms showed an 11% decrease in AUC for fluorescence area and a 32% decrease in AUC for fluorescence intensity in the three patients whose conditions clinically improved but increases of 131% and 292% in the three patients whose conditions clinically worsened. In group 2, a 38% decrease in AUC for fluorescence intensity and a 19% decrease in AUC for fluorescence area were observed in patients who received VEGF Trap compared with increases of 66% (P = 0.004, Mann-Whitney U test) and 21% (P = 0.07) for patients who received placebo. Macular volume decreased by 11% in VEGF Trap-treated patients and increased by 10% in placebo-treated patients (P = 0.03). CONCLUSIONS: This study reports a technique for analysis of change in fluorescence area and intensity over time during fluorescein angiography (FA) using a continuous scale and its application in a clinical setting and a clinical trial. Compared with previous techniques making use of categorical scales, this approach provides an advantage for evaluating responses to treatment that may improve the value of FA as an outcome measure in clinical trials. PMID- 17122138 TI - Calpain-specific proteolysis in primate retina: Contribution of calpains in cell death. AB - PURPOSE: One of the leading causes of blindness is retinal damage caused by the high intraocular pressure (IOP) in glaucoma. Previous studies in rats have suggested that the proteolytic enzyme calpain (EC 3.4.22.17) is involved in retinal cell death during ischemia and in acute high IOP. Ubiquitous, calcium activated calpain-1 and -2 from monkey retina are highly homologous to rat calpains, although expression patterns in variants of tissue-specific calpain-3 are different between monkey and rodent retinas. Thus, the purpose of the present study was to investigate the involvement of calpain-induced proteolysis in retinal cell death in primates. METHODS: Calpain involvement in a simulated pathologic condition was examined by incubating monkey retinas in hypoxic conditions (95% N2 and 5% CO2) in RPMI medium without glucose. Endogenous tissue calpains were also directly activated in monkey and human retinal soluble proteins by incubating with 2.5 mM calcium. The resultant proteolysis of monkey retinal proteins was assessed by 2D electrophoresis (2-DE). RESULTS: In hypoxic retina, leakage of lactate dehydrogenase (LDH) from retinas into the medium increased, indicating cell death. LDH leakage was partially inhibited by the calpain inhibitor SJA6017. Calpain autolysis was observed, and the calpain preferred substrate alpha-spectrin was proteolyzed. In retinal soluble proteins incubated with calcium, a total of 15 spots from 2-DE of retinal soluble proteins were identified by mass spectrometry. Proteolysis of major proteins, vimentin, beta-tubulin, alpha-enolase, and Hsp70 were confirmed by immunoblot analysis. Activation of calpains and proteolysis of these substrates were inhibited by the calpain-specific inhibitor SJA6017. CONCLUSIONS: Taken together, these results suggested that calpain activation in primate retinas could play an important role in cell death during hypoxia caused by elevated IOP from glaucoma. PMID- 17122139 TI - Chronic ingestion of (3R,3'R,6'R)-lutein and (3R,3'R)-zeaxanthin in the female rhesus macaque. AB - PURPOSE: To investigate how supplementation of the monkey's diet with high doses of lutein (L), zeaxanthin (Z), or a combination of the two affects the plasma levels and ocular tissue deposition of these carotenoids and their metabolites over time and to determine whether these high doses can cause ocular toxicity. METHODS: Eighteen female rhesus monkeys were divided into groups of control (n = 3 control), L-treated (n = 5, 9.34 mg lutein/kg and 0.66 mg zeaxanthin/kg), Z treated (n = 5, 10 mg zeaxanthin/kg), and L/Z-treated (n = 5, lutein and zeaxanthin, each 0.5 mg/kg). After 12 months of daily supplementation, one control animal, two L-treated animals, two Z-treated animals, and all the L/Z treated animals were killed. The rest of the monkeys were killed after an additional six months without supplementation. Plasma and ocular tissue carotenoid analyses, fundus photography, and retina histopathology were performed on the animals. RESULTS: Supplementation of monkeys with L and/or Z increased the mean plasma and ocular tissue concentrations of these carotenoids and their metabolites. The mean levels of L and Z in the retinas of the L- and Z-treated animals after 1 year increased significantly over baseline. High dose supplementation of monkeys with L or Z did not cause ocular toxicity and had no effect on biomarkers associated with kidney toxicity. CONCLUSIONS: The mean levels of L and Z in plasma and ocular tissues of the rhesus monkeys increase with supplementation and in most cases correlate with the levels of their metabolites. Supplementation of monkeys with L or Z at high doses, or their combination does not cause ocular toxicity. PMID- 17122140 TI - Human macula investigated in vivo with polarization-sensitive optical coherence tomography. AB - PURPOSE: To investigate a depolarizing layer that is visible in polarization sensitive optical coherence tomography (PS-OCT) images of the retina. To identify this layer and characterize its depolarizing effect quantitatively. METHODS: Ten healthy human subjects (mean age, 31 +/- 8 years) and two patients with RPE diseases participated in the study. The macular region of one eye of each subject was investigated with a phase-resolved PS-OCT system. The instrument measured backscattered intensity (standard OCT), phase retardation, and (cumulative) birefringent axis orientation, simultaneously. For a quantification of the depolarizing layer, plots of the distributions of retardation and axis orientation within and above this layer were analyzed. RESULTS: A polarization scrambling layer (PSL) was observed at the posterior boundary of the retina in PS OCT images of all volunteers. It was identified in PS-OCT images by determining random retardation and axis orientation in a transverse direction. Measurements in patients with neurosensory retinal detachment, retinal pigment epithelium (RPE) detachment, and RPE atrophy suggest that the PSL is the RPE. The statistical analysis provided objective discrimination of the RPE from the other retinal structures. CONCLUSIONS: PS-OCT represents a powerful tool for increasing image contrast in ocular tissues. The observed polarization-scrambling nature of the RPE may be used in diseased eyes to locate the RPE or remains of the RPE definitively in OCT images. PMID- 17122141 TI - Autofluorescence characteristics of early, atrophic, and high-risk fellow eyes in age-related macular degeneration. AB - PURPOSE: To assess the relationships of drusen, pigment, and focally increased autofluorescence (FIAF) and the reticular pattern of hypoautofluorescence, to distinguish the combined photographic and AF characteristics of early, atrophic, and high-risk fellow eyes in AMD. METHODS: In a retrospective interinstitutional clinical study, AF and color photograph pairs of 221 eyes were examined: 166 eyes of 83 patients with bilateral large, soft drusen, with and without geographic atrophy (GA), and 55 fellow eyes of 55 patients with unilateral choroidal neovascularization (CNV). Forty-two eyes (one eye from each of 42 patients with early or atrophic AMD) were divided into four groups: 14 with drusen only, 9 with drusen and pigment abnormalities, 11 fellow eyes of patients with unilateral GA, and 8 eyes of patients with bilateral GA (acronyms for the groups: D-D, D-Pig, D GA and GA-GA, respectively). The 55 fellow eyes of patients with CNV were divided into three groups: 19 eyes with no FIAF (CNV-0), 16 with FIAF without reticular AF (CNV-1), and 20 eyes with reticular AF and/or pseudodrusen (CNV-R). Image pairs of eyes with FIAF were registered, and drusen, pigment, and FIAF were segmented using automated background leveling and thresholding. All 221 eyes were surveyed for reticular AF and reticular pseudodrusen. The main outcome measures were (1) the fraction and relative probability of FIAF colocalizing with drusen and pigment and (2) the presence or absence of reticular AF and reticular pseudodrusen. RESULTS: The mean fractions of FIAF that colocalized with large drusen were: D-D group, 0.46 +/- 0.21; D-Pig group, 0.42 +/- 0.29; D-GA group, 0.13 +/- 0.09; and GA-GA group, 0.11 +/- 0.12. Comparisons between groups showed significant differences when comparing either the D-D group or the D-Pig group with either the D-GA group or the GA-GA group (P between 0.0001 and 0.015), whereas other comparisons were nonsignificant (Mann-Whitney rank sum test). The mean probabilities of FIAF colocalizing with large drusen relative to chance (1.0) were: D-D group, 4.7 +/- 2.5; D-Pig group, 4.3 +/- 2.3; D-GA group, 1.4 +/- 0.8; and GA-GA group, 1.8 +/- 1.3, with similar significant differences as for the colocalization fractions. The mean probability of FIAF colocalizing with small to intermediate drusen in the D-D group was 1.5 +/- 1.3, which was not significantly different from chance. In the D-Pig group, the median probability of FIAF colocalizing with pigment abnormalities was 10.0 (range, 1.1-51.0). The AF patterns in 15 of 19 eyes in the CNV-0 group were normal; the remainder had nonreticular hypoautofluorescence only. In the CNV-1 group, the relations of FIAF with drusen and pigment were similar to those in the early AMD groups. CNV-R comprised 20 of 55 eyes in the CNV group, but reticular autofluorescence and/or pseudodrusen were found in only 14 of 166 eyes of the early and atrophic groups. Of the 34 total eyes with reticular AF or pseudodrusen, 28 had both, 4 had reticular AF only, and 2 had reticular pseudodrusen only. CONCLUSIONS: There are clear relationships between AF patterns and clinical AMD status. In early AMD, FIAF's colocalization with large, soft drusen and hyperpigmentation is several times greater than chance, suggesting linked disease processes. In advanced atrophic AMD, FIAF is found mostly adjacent to drusen and GA, suggesting that dispersal of drusen-associated lipofuscin is a marker of atrophic disease progression. In the neovascular case, a large group of fellow eyes have no FIAF abnormalities, suggesting that lipofuscin is not a major determinant of CNV. However, reticular hypoautofluorescence, consistent with widespread inflammatory damage to the RPE, appears to be a highly sensitive imaging marker for the disease that determines reticular pseudodrusen and is strongly associated with CNV. PMID- 17122142 TI - CTRP5 is a membrane-associated and secretory protein in the RPE and ciliary body and the S163R mutation of CTRP5 impairs its secretion. AB - PURPOSE: In a prior study, a S163R mutation in the complement-1q tumor necrosis factor-related protein 5 (CTRP5/ C1QTNF5) was reported to be associated with early-onset long anterior zonules (LAZ) and late-onset retinal degeneration (L ORD). The ocular tissues involved in the phenotype are the retinal pigment epithelium (RPE) in the posterior segment and ciliary epithelium (CE) and lens in the anterior segment. The purpose of this study was to characterize the spatial and temporal expression of the mouse Ctrp5 gene, determine tissue and subcellular localization, and study the effect of the S163R mutation. METHODS: The expression of the Ctrp5 gene in the mouse was studied by quantitative (q)RT-PCR and in situ hybridization. CTRP5 protein expression and distribution were studied by Western blot analysis, immunohistochemistry, and immunoelectron microscopy. Cellular location of wild-type and mutant CTRP5 in MDCK and COS-7 cells was determined by immunofluorescence and immunoblot analysis. RESULTS: A significant level of Ctrp5 expression was detected in the adult mouse in the ciliary body (CB) and RPE, and expression started at a very early stage of embryogenesis. Immunohistochemical analysis showed CTRP5 protein in the apical processes of the RPE and forming a hexagonal lattice associated with the RPE lateral membranes. In the ciliary body, CTRP5 was localized to the apical aspects of the CE, the region between the bilayered ciliary epithelial cells. The membrane association of CTRP5 in the RPE and CE was further confirmed by immunoelectron microscopy. Furthermore, cultured cells were used to show that the CTRP5 is a secretory protein and that its secretion is impaired by the S163R mutation. CONCLUSIONS: CTRP5, a secretory and membrane-associated protein, is localized to the lateral and apical membranes of the RPE and CB. Impaired secretion of the mutant protein may underlie the pathophysiology of L-ORD and LAZ. PMID- 17122143 TI - Spatial and temporal expression of MFRP and its interaction with CTRP5. AB - PURPOSE: Mutations in the membrane frizzled-related protein (MFRP) gene cause nanophthalmos in humans, and a splice site mutation causes recessive retinal degeneration in the rd6 mouse. In human and mouse genomes, the MFRP gene lies adjoining to the complement 1q tumor necrosis factor-related protein 5 (CTRP5/C1QTNF5) gene involved in causing retinal degeneration and abnormal lens zonules in human. The purpose of this study was to characterize the spatial and temporal expression of the mouse Mfrp gene, determine tissue and subcellular localization of MFRP protein, and study its interaction with CTRP5. METHODS: Expression of the Mfrp gene in the mouse was studied by quantitative (q)RT-PCR. MFRP protein expression and distribution were studied by Western blot analysis, immunohistochemistry, and immunoelectron microscopy. Interaction with CTRP5 was studied by immunoprecipitation and immunoblot analysis, using mouse eye and human retinal pigmented epithelium (RPE) choroid extracts and by expressing full-length CTRP5 and MFRP in a heterologous system. RESULTS: The Mfrp gene is specifically expressed in RPE and ciliary body (CB), and its expression starts during early stages of embryogenesis. In the albino mouse eye, MFRP is localized to the apical and basal membranes of RPE and ciliary epithelium (CE). In addition, MFRP and CTRP5 were found to colocalize in RPE, CE, and MDCK cells, a general model of polarized epithelia. These proteins interact with each other in ocular tissues and also in a heterologous system. CONCLUSIONS: MFRP is localized to the plasma membrane of CE and RPE, and colocalizes and interacts with CTRP5 indicating a functional relationship between these two proteins. PMID- 17122144 TI - Noninvasive volumetric imaging and morphometry of the rodent retina with high speed, ultrahigh-resolution optical coherence tomography. AB - PURPOSE: To demonstrate high-speed, ultrahigh-resolution optical coherence tomography (OCT) for noninvasive, in vivo, three-dimensional imaging of the retina in rat and mouse models. METHODS: A high-speed, ultrahigh-resolution OCT system using spectral, or Fourier domain, detection has been developed for small animal retinal imaging. Imaging is performed with a contact lens and postobjective scanning. An axial image resolution of 2.8 mum is achieved with a spectrally broadband superluminescent diode light source with a bandwidth of approximately 150 nm at approximately 900-nm center wavelength. Imaging can be performed at 24,000 axial scans per second, which is approximately 100 times faster than previous ultrahigh-resolution OCT systems. High-definition and three dimensional retinal imaging is performed in vivo in mouse and rat models. RESULTS: High-speed, ultrahigh-resolution OCT enabled high-definition, high transverse pixel density imaging of the murine retina and visualization of all major intraretinal layers. Raster scan protocols enabled three-dimensional volumetric imagingand comprehensive retinal segmentation algorithms allowed measurement of retinal layers. An OCT fundus image, akin to a fundus photograph was generated by axial summation of three-dimensional OCT data, thus enabling precise registration of OCT measurements to retinal fundus features. CONCLUSIONS: High-speed, ultrahigh-resolution OCT enables imaging of retinal architectural morphology in small animal models. OCT fundus images allow precise registration of OCT images and repeated measurements with respect to retinal fundus features. Three-dimensional OCT imaging enables visualization and quantification of retinal structure, which promises to allow repeated, noninvasive measurements to track disease progression, thereby reducing the need for killing the animal for histology. This capability can accelerate basic research studies in rats and mice and their translation into clinical patient care. PMID- 17122145 TI - Time-lapse imaging of vitreoretinal angiogenesis originating from both quiescent and mature vessels in a novel ex vivo system. AB - PURPOSE: Diabetic retinopathy (DR) is an angiogenic disease that leads to severe visual loss. However, adequate animal models of vitreoretinal neovascularization in proliferative diabetic retinopathy (PDR) have not yet been described. The purpose of this study was to develop a novel ex vivo system for assessing vitreoretinal angiogenic processes that originate from both quiescent and mature vessels that could be observed with time-sequential imaging. METHODS: The retinas of 7- to 8-week-old mice were cultured for 4 days, with or without several growth factors with novel procedures, and immunohistochemistry was performed. The retinas from Tie2-GFP mice were cultured with vascular endothelial growth factor (VEGF), and time-sequential imaging of vitreoretinal angiogenesis was acquired. RESULTS: Vascular sprouts were induced by both VEGF and placenta growth factor, but not by insulin-like growth factor-1, basic fibroblast growth factor or angiopoietin-2. In explants with or without VEGF, perivascular mural cells were dissociated from endothelial cells, which is an important step during angiogenesis and in the progression of DR. Furthermore, use of time-lapse observations of retinal neovascularization events visualized that the first step in vascular sprout emergence from quiescent vessels was a single cell extension. The leading edges of a sprouting endothelial cell extended and retracted in a sequential manner. From newly formed vessels, additional vascular sprouts then emerged and new vessels fused to each other, resulting in vascular branching. CONCLUSIONS: Time-lapse imaging of this system visualized the dynamic process in vitreoretinal neovascularization from quiescent and mature vessels. PMID- 17122146 TI - Retinal pigment epithelial cells synthesize laminins, including laminin 5, and adhere to them through alpha3- and alpha6-containing integrins. AB - PURPOSE: Retinal diseases are often accompanied by changes in the structure of the multilayered extracellular matrix underlying the retina, Bruch's membrane (BrM). These structural revisions potentially lead to alterations in retinal pigment epithelium (RPE) adhesion, likely via modification of interactions with extracellular matrix (ECM) proteins including laminins in BrM. The purpose of this study was to identify specific laminins in BrM and their receptors in RPE cells. METHODS: The laminin composition of BrM was determined using biochemical, molecular biological, and immunohistochemical techniques of rat, bovine, and human tissue and cell lines. An adhesion assay was used to test RPE attachment to laminins and the receptors used for this attachment. RESULTS: BrM contained laminin chains that could form laminin heterotrimers including laminins 1, 5, 10, and 11. RPE cells synthesized these laminin chains in vitro. Therefore, RPE cells may synthesize BrM laminins. The RPE cells preferentially adhered to potential BrM laminins. Although the cells adhered to the BrM component collagen IV, these cells preferentially adhered to laminins. Of the laminins tested, the RPE cells adhered preferentially to laminin 5. The cells interacted with these laminins via specific integrins and attained a different morphology on each laminin. In particular, the RPE cells rapidly attached and flattened on laminin 5. CONCLUSIONS: BrM contains specific laminins, and RPE cells express integrin receptors for those laminins. The interaction of these specific laminins and integrins most likely leads to differential behavior of RPE cells. PMID- 17122147 TI - Neuroprotective effects of angiotensin II type 1 receptor (AT1R) blocker, telmisartan, via modulating AT1R and AT2R signaling in retinal inflammation. AB - PURPOSE: To investigate the retinal neural damage that occurs during inflammation and the therapeutic effects of the angiotensin II type 1 receptor (AT1R) blocker, telmisartan, using a model of endotoxin-induced uveitis (EIU). METHODS: The localization of AT1R and AT2R was shown by immunohistochemistry. EIU was induced by intraperitoneal injection of lipopolysaccharide (LPS). Animals were treated with telmisartan for 2 days and were evaluated 24 hours later. Expression levels of angiotensin II, STAT3 activation induced by inflammatory cytokines, and retinal proteins essential for neural activities (e.g., synaptophysin, rhodopsin) were analyzed by immunoblot. An AT2R antagonist was administered to evaluate the contribution of AT2R signaling in this therapy. Dark-adapted full-field electroretinography (ERG) was also performed. RESULTS: AT1R and AT2R were expressed in presynaptic terminals in most of the retinal neurons. AT1R was also expressed in Muller glial cells. During inflammation, angiotensin II expression was elevated, STAT3 was activated, and synaptophysin and rhodopsin expression were reduced. The expression of glial fibrillary acidic protein (GFAP), downstream of STAT3 activation, was induced in Muller glial cells. However, treatment with telmisartan successfully avoided all these changes. An AT2R antagonist lowered synaptophysin expression despite the treatment. STAT3 activity was negatively correlated with rhodopsin expression. Furthermore, ERG responses, which were mostly prevented by telmisartan, were disturbed during inflammation. CONCLUSIONS: Retinal protein expression and visual function are both disturbed by inflammation. Treatment with the AT1R blocker telmisartan efficiently prevented these signs of retinal neural damage through the reduction of local angiotensin II expression, the blockade of AT1R, and the relative upregulation of AT2R function. PMID- 17122148 TI - Rod photoreceptor loss in Rho-/- mice reduces retinal hypoxia and hypoxia regulated gene expression. AB - PURPOSE: This study was conducted to evaluate whether regions of the retinal neuropile become hypoxic during periods of high oxygen consumption and whether depletion of the outer retina reduces hypoxia and related changes in gene expression. METHODS: Retinas from rhodopsin knockout (Rho-/-) mice were evaluated along with those of wild-type (WT) control animals. Retinas were also examined at the end of 12-hour dark or light periods, and a separate group was treated with l cis-diltiazem at the beginning of a 12-hour dark period. Hypoxia was assessed by deposition of hypoxyprobe (HP) and HP-protein adducts were localized by immunohistochemistry and quantified using ELISA. Also, hypoxia-regulated gene expression and transcriptional activity were assessed alongside vascular density. RESULTS: Hypoxia was observed in the inner nuclear and ganglion cell layers in WT retina and was significantly reduced in Rho-/- mice (P < 0.05). Retinal hypoxia was significantly increased during dark adaptation in WT mice (P < 0.05), whereas no change was observed in Rho-/- or with l-cis-diltiazem-treated WT mice. Hypoxia inducible factor (HIF)-1alpha DNA-binding and VEGF mRNA expression in Rho-/- retina was significantly reduced in unison with outer retinal depletion (P < 0.05). Retina from the Rho-/- mice displayed an extensive intraretinal vascular network after 6 months, although there was evidence that capillary density was depleted in comparison with that in WT retinas. CONCLUSIONS: Relative hypoxia occurs in the inner retina especially during dark adaptation. Photoreceptor loss reduces retinal oxygen usage and hypoxia which corresponds with attenuation of the retinal microvasculature. These studies suggest that in normal physiological conditions and diurnal cycles the adult retina exists in a state of borderline hypoxia, making this tissue particularly susceptible to even subtle reductions in perfusion. PMID- 17122149 TI - Retinopathy is reduced during experimental diabetes in a mouse model of outer retinal degeneration. AB - PURPOSE: Diabetic patients who also have retinitis pigmentosa (RP) appear to have fewer and less severe retinal microvascular lesions. Diabetic retinopathy may be linked to increased inner retinal hypoxia, with the possibility that this is exacerbated by oxygen usage during the dark-adaptation response. Therefore, patients with RP with depleted rod photoreceptors may encounter proportionately less retinal hypoxia, and, when diabetes is also present, there may be fewer retinopathic lesions. This hypothesis was tested in rhodopsin knockout mice (Rho /-) as an RP model in which the diabetic milieu is superimposed. The study was designed to investigate whether degeneration of the outer retina has any impact on hypoxia, to examine diabetes-related retinal gene expression responses, and to assess lesions of diabetic retinopathy. METHODS: Streptozotocin-induced diabetes was created in male C57Bl6 (wild-type; WT) and Rho-/- mice, and hyperglycemia was maintained for 5 months. The extent of diabetes was confirmed by measurement of glycated hemoglobin (%GHb) and accumulation of advanced glycation end products (AGEs). Retinal hypoxia was assessed using the bioreductive drug pimonidazole. The retinal microvasculature was studied in retinal flatmounts stained by the ADPase reaction, and the outer retina was evaluated histologically in paraffin embedded sections. Retinal gene expression of VEGF-A, TNF-alpha, and mRNAs encoding basement membrane component proteins were quantified by real-time RT PCR. RESULTS: The percentage GHb increased significantly in the presence of diabetes (P < 0.001) and was not different between WT or Rho-/- mice. Hypoxia increased in the retina of WT diabetic animals when compared with controls (P < 0.001) but this diabetes-induced change was absent in Rho-/- mice. Retinal gene expression of VEGF-A was significantly increased in WT mice with diabetes (P < 0.05), but was unchanged in Rho-/- mice. TNF-alpha gene expression significantly increased (4.9-fold) in WT mice with diabetes (P < 0.05) and also increased appreciably in Rho-/- mice but to a reduced extent (1.5 fold; P < 0.05). The outer nuclear layer in nondiabetic Rho-/- mice was reduced to a single layer after 6 months, but when diabetes was superimposed on this model, there was less degeneration of photoreceptors (P < 0.05). Vascular density was attenuated in diabetic WT mice compared with the nondiabetic control (P < 0.001); however, this diabetes-related disease was not observed in Rho-/- mice. CONCLUSIONS: Loss of the outer retina reduces the severity of diabetic retinopathy in a murine model. Oxygen usage by the photoreceptors during dark adaptation may contribute to retinal hypoxia and exacerbate the progression of diabetic retinopathy. PMID- 17122150 TI - Activation of caspase-8 and caspase-12 pathways by 7-ketocholesterol in human retinal pigment epithelial cells. AB - PURPOSE: To determine whether caspase or cathepsin pathways are activated in human retinal pigment epithelial cells (ARPE-19) after exposure to 7 ketocholesterol (7kCh). METHODS: ARPE-19 cells were exposed to 7kCh with or without z-VAD-fmk, a pan-caspase inhibitor. Caspase-3, -8, and -9 activities were measured by a fluorochrome inhibitor of caspase (FLICA) assay. Caspase-12 activity was detected by Western blotting. RT-PCR was performed for 18s, mortalin 2, cathepsins B, D, and L/V2. RESULTS: At 24 hours, 7kCh-treated cultures had increased caspase-8 (P < 0.001) and caspase-3 (P < 0.001) activities compared with vehicle-treated cultures. 7kCh-induced caspase-3 activation was blocked by z VAD-fmk (P < 0.001). Caspase-9 was not activated by 7kCh treatment (P > 0.05). Procaspase-12 was cleaved into its active form after treatment with 7kCh for 24 hours. At 6 hours, the RNA level for mortalin-2, a pro-survival gene, was upregulated. ARPE-19 cells did not express RNA for cathepsins B, D, or L/V2 under any conditions. CONCLUSIONS: In ARPE-19 cells, 7kCh-induced apoptosis uses the receptor-mediated caspase-8 pathway and the endoplasmic reticulum stress-induced caspase-12 pathway but not the mitochondrial caspase-9 pathway. The cathepsin pathways are not involved in 7kCh-induced cell death. These data demonstrate that 7kCh causes a loss of cell viability through caspase-dependent apoptosis and can act as an oxidative stressor leading to retinal pigment epithelial cell atrophy. Elucidating the specific apoptotic pathways involved may have therapeutic potential for AMD and other retinal diseases. PMID- 17122151 TI - Expression, subcellular localization, and regulation of sigma receptor in retinal muller cells. AB - PURPOSE: Sigma receptors (sigmaRs) are nonopioid, nonphencyclidine binding sites with robust neuroprotective properties. Type 1 sigmaR1 (sigmaR1) is expressed in brain oligodendrocytes, but its expression and binding capacity have not been analyzed in retinal glial cells. This study examined the expression, subcellular localization, binding activity, and regulation of sigmaR1 in retinal Muller cells. METHODS: Primary mouse Muller cells (MCs) were analyzed by RT-PCR, immunoblotting, and immunocytochemistry for the expression of sigmaR1, and data were compared with those of the rat Muller cell line (rMC-1) and the rat ganglion cell line (RGC-5). Confocal microscopy was used to determine the subcellular sigmaR1 location in primary mouse MCs. Membranes prepared from these cells were used for binding assays with [3H]-pentazocine (PTZ). The kinetics of binding, the ability of various sigmaR1 ligands to compete with sigmaR1 binding, and the effects of donated nitric oxide (NO) and reactive oxygen species (ROS) on binding were examined. RESULTS: sigmaR1 is expressed in primary mouse MCs and is localized to the nuclear and endoplasmic reticulum membranes. Binding assays showed that in primary mouse MCs, rMC-1, and RGC-5, the binding of PTZ was saturable. [3H]-PTZ bound with high affinity in RGC-5 and rMC-1 cells, and the binding was similarly robust in primary mouse MCs. Competition studies showed marked inhibition of [3H]-PTZ binding in the presence of sigmaR1-specific ligands. Incubation of cells with NO and ROS donors markedly increased sigmaR1 binding activity. CONCLUSIONS: MCs express sigmaR1 and demonstrate robust sigmaR1 binding activity, which is inhibited by sigmaR1 ligands and is stimulated during oxidative stress. The potential of Muller cells to bind sigmaR1 ligands may prove beneficial in retinal degenerative diseases such as diabetic retinopathy. PMID- 17122152 TI - Loss of BCL-XL in rod photoreceptors: Increased susceptibility to bright light stress. AB - PURPOSE: BCL-X(L), an anti-apoptotic member of the BCL-2 family proteins and a cell death/survival checkpoint regulator, was shown to be upregulated in bright light-stressed mouse photoreceptors during an investigation of bright light induced protein expression. To investigate the significance of BCL-X(L) upregulation in the bright light damage model, the Bcl-x gene was disrupted specifically in mouse rod photoreceptors, and the effect of Bcl-x disruption was characterized on retinal apoptosis, function, and morphology. METHODS: Rod specific Bcl-x knockout mice, generated by mating mouse opsin promoter-controlled Cre mice with floxed Bcl-x mice, were subjected to bright light stress. Retinal apoptosis in the bright light-stressed conditional Bcl-x knockout mice was characterized with TUNEL, DNA fragmentation, and nuclear staining assays. Photoreceptor structural and functional integrity in the bright light-stressed conditional Bcl-x knockout mice was determined by measuring photoreceptor outer nuclear layer (ONL) thickness and electroretinography amplitudes. RESULTS: Disruption of Bcl-x in rod photoreceptors caused increased photoreceptor apoptosis, decreased retinal function, and decreased ONL thickness in bright light-stressed mice. CONCLUSIONS: The loss of BCL-X(L) increased rod photoreceptor susceptibility to bright light stress. Although the biochemical mechanism(s) of BCL-X(L) in photoreceptor death or survival has not been investigated extensively, results of the present study suggest that BCL-X(L), a cell survival/death checkpoint regulator, is involved in photoreceptor survival under bright light stress. PMID- 17122153 TI - Cell cycle-specific and cell type-specific expression of Rb in the developing human retina. AB - PURPOSE: To define the pattern of Rb expression relative to cell cycle position and cell type in the developing human retina. METHODS: Cryosections of fetal week 11-18 retinas were immunostained for Rb and cell cycle- or cell type-specific markers. RESULTS: Rb was prominent in retinal progenitor cells (RPCs) expressing the cyclin D1, cyclin A, and cytoplasmic cyclin B markers of G1, S, and early to mid G2 phases, but not in RPCs expressing the phosphohistone H3 marker of late G2 and M. Rb was not detected in the earliest postmitotic ganglion, amacrine, horizontal, and bipolar cell precursors migrating away from the ventricular layer, but was detected as such cells underwent further differentiation. Among photoreceptors, Rb was not detected in the earliest RXRgamma(+) cone precursors or in the earliest Nrl(+) rod precursors, but subsequently rose to high levels in cones and to low levels in rods. Rb was prominent at the time when Muller glia exit the cell cycle and was generally expressed in a pattern complementary to p27(Kip1). CONCLUSIONS: Rb exhibits cell cycle-specific expression in RPCs, with loss in late G2-M and restoration in G1. Rb is re-expressed after postmitotic ganglion, amacrine, horizontal, and bipolar cell precursors migrate away from the ventricular layer; after the appearance of early cone and rod markers; but coinciding with Muller glia cell cycle withdrawal. The results suggest that Rb does not mediate the initial proliferative arrest of retinal neurons, but may indirectly induce arrest in RPCs or maintain an arrest in postmitotic precursors. PMID- 17122154 TI - Orientation discrimination and contrast detection thresholds in migraine for cardinal and oblique angles. AB - PURPOSE: To determine whether orientation discrimination deficits in migraine, which have been found to depend on the spatial frequency of the stimulus, are due to precortical dysfunction or to abnormal patterns of orientation tuning at cortical loci. Further, to assess whether any cortical involvement is restricted to the striate cortex or whether higher cortical areas are also involved. Orientation-specific abnormalities would provide evidence of cortical dysfunction. METHODS: Orientation-discrimination and contrast-detection thresholds were assessed at cardinal (0 degrees) and oblique (45 degrees) orientations using explicit lines defined by Gabor patches. To test for extrastriate dysfunction, participants made orientation judgments using virtual lines defined by two widely spaced circles. Migraine history, migraine triggers, and pattern sensitivity were also assessed. Twenty migraineurs (10 with visual aura, 10 without) and 20 control participants were tested. RESULTS: Orientation discrimination thresholds were lower for discriminations made about the cardinal axis than for discriminations made about the oblique axis, a well-documented phenomenon known as the oblique effect. Relative to the control group, the migraine group exhibited orientation-specific sensitivity losses on explicit and virtual judgments. Orientation-discrimination thresholds about the oblique axis were significantly elevated in the migraine group. In contrast, the migraine and control groups' detection thresholds did not differ. CONCLUSIONS: These findings reflect abnormal function of striate and extrastriate cortex in migraine. In addition, the discrimination data are consistent with wider orientation-tuning curves for orientation-sensitive cells in migraine, whereas the detection data suggest peak sensitivity does not differ between the groups. PMID- 17122155 TI - Contrasting blue-on-yellow with white-on-white visual fields: Roles of visual adaptation for healthy peri- or postmenopausal women younger than 70 years of age. AB - PURPOSE: To test the hypothesis that differences between short-wavelength automated perimetry (SWAP) and white-on-white visual field sensitivities are related to between-individual variation in the visual adaptation properties of SWS cone pathways. METHODS: Twenty-six healthy amenorrheic (peri- or postmenopausal) women not using hormonal medication were tested. Subjects ranged in age from 48 to 68 years. They were tested by using foveal increment-threshold techniques and also with two types of 24-2 visual field tests: a full-threshold SWAP blue-on-yellow (B/Y) test and a white-on-white (W/W) test obtained using a Swedish Interactive Threshold Algorithm (SITA Standard). The age-corrected sensitivity differences between the two types of visual fields were compared against foveal measures of visual sensitivity and adaptation, which were obtained psychophysically using dim and bright yellow backgrounds. All measurements for each subject were made at a single testing session. The comparisons were made for the entire visual field and for separate portions of the visual field. The analyses also included pupil size data obtained during visual field testing. RESULTS: The B/Y minus W/W (B/Y - W/W) mean deviation difference was described (R = 0.80) by a multilinear model with three significant factors: (1) an adaptation factor and (2) a baseline sensitivity factor, each derived from the foveal psychophysical data for short-wavelength test stimuli, and (3) a pupil size factor, as recorded for SWAP. The total deviation differences in the periphery of the visual field (approximately 22 degrees from fixation) were described (R = 0.87) by a model with four significant factors, the fourth being an "eccentricity factor" describing the rate of change of the B/Y - W/W total deviation difference measured as a function of increasing retinal eccentricity approximately 9 degrees -17 degrees from fixation. More than 40% of the variance in the B/Y - W/W mean deviation differences was accounted for either directly or indirectly (via effects of pupil size) by variations in adaptation to the yellow background used for SWAP. CONCLUSIONS: Much of the extra variability in SWAP sensitivities for a select group of healthy women can be accounted for by differences in the degree of desensitization induced by the yellow background used for SWAP. For clinical practice, pupil status (dilated or undilated) should be altered only with caution from one SWAP testing session to another. PMID- 17122156 TI - Emerging mechanisms of immunosuppression in oral cancers. AB - Mounting effective anti-tumor immune responses against tumors by both the innate and adaptive immune effectors is important for the clearance of tumors. However, accumulated evidence indicates that immune responses that should otherwise suppress or eliminate transformed cells are themselves suppressed by the function of tumor cells in a variety of cancer patients, including those with oral cancers. Signaling abnormalities, spontaneous apoptosis, and reduced proliferation and function of circulating natural killer cells (NK), T-cells, dendritic cells (DC), and tumor-infiltrating lymphocytes (TILs) have been documented previously in oral cancer patients. Several mechanisms have been proposed for the functional deficiencies of tumor-associated immune cells in oral cancer patients. Both soluble factors and contact-mediated immunosuppression by the tumor cells have been implicated in the inhibition of immune cell function and the progression of tumors. More recently, elevated levels and function of key transcription factors in tumor cells, particularly NFkappaB and STAT3, have been shown to mediate immune suppression in the tumor microenvironment. This review will focus on these emerging mechanisms of immunosuppression in oral cancers. PMID- 17122157 TI - Tissue inhibitors of metalloproteinases (TIMPs): their biological functions and involvement in oral disease. AB - Several families of enzymes are responsible for the degradation of extracellular matrix (ECM) proteins during the remodeling of tissues. An important family of such enzymes is that of the matrix metalloproteinases (MMPs). To control MMP mediated ECM breakdown, tissue inhibitors of metalloproteinases (TIMPs) are able to inhibit MMP activity. A disturbed balance of MMPs and TIMPs is found in various pathologic conditions, such as cancer, rheumatoid arthritis, and periodontitis. The role of MMPs in pathology has been extensively described in the literature. The main focus of this review lies in the biological functions of TIMPs and their occurrence in disease, especially in the head and neck area. Their biological functions and their role in diseases like oral cancers and periodontitis, and in the development of cleft palate, will be discussed. Finally, the diagnostic and therapeutical opportunities of TIMPs will be evaluated. PMID- 17122158 TI - Preventive intervention possibilities in radiotherapy- and chemotherapy- induced oral mucositis: results of meta-analyses. Authors failed to acknowledge the Cochrane review on this topic. PMID- 17122159 TI - Influence of alveolar support on stress in periodontal structures. AB - The influence of alveolar bone support on the functional capability of a tooth remains unclear. It was hypothesized that a reduction in alveolar support causes an increase of maximum stress in the periodontal structures. Mathematical models of the maxillary incisor to simulate in vivo tooth movement were constructed with periodontium of normal or reduced bone height, and normal or widened periodontal ligament (PDL) space. Under simulated bite force, the maximum tensile stress at the lingual cervical region in the PDL increased with bone height reduction, but decreased with PDL widening. The compressive stress at the cervical region in the cortical bone was no more than 22% of the yield strength of bone, and did not increase by the height reduction with widened PDL. The result suggests that the height reduction potentially causes mechanical damage to the PDL, but, of itself, is not likely to have a negative effect on the bone. PMID- 17122160 TI - Electric-current-assisted application of self-etch adhesives to dentin. AB - The use of electric current during the application of etch-and-rinse adhesive systems has been recently claimed to increase bonding of etch-and-rinse adhesives by enhancing substrate impregnation. The null hypothesis tested in this study was that electrically assisted application has no effect on bond strength of self etching bonding systems. Three self-etch adhesives (Protect-Bond, Xeno III, and Prompt L-Pop) were applied with the aid of an electric signal-generating device (ElectroBond) and tested vs. controls prepared with the same disposable sponges but without electric current. Specimens bonded under the influence of electric current exhibited increased microtensile bond strength compared with the controls (p<0.05). High-resolution SEM analysis showed that bonding under the influence of electricity reduced interfacial nanoleakage. It is speculated that resin infiltration may be improved by the attraction of polar monomers by an electric current or by modification of the dentin surface charges, resulting in better water substitution or evaporation. PMID- 17122161 TI - Antagonist enamel wears more than ceramic inlays. AB - Wear phenomena of ceramic inlays are not fully understood. The aim of the present study was to evaluate ceramic wear, antagonist enamel wear, and luting cement wear over 8 years. The two-fold null hypothesis was that there would be (1) no difference in wear behavior between ceramic and enamel, and (2) no influence of filler content of luting composites on composite wear. From 96 restorations, 36 Class II inlays from 16 participants were selected. For inlays with opposing enamel cusps (n=17), replicas of inlays and enamel were scanned with a 3-D laser scanner. Luting gaps of inlays (n=36) were analyzed with a profilometer, including 3-D data analysis. Ceramic and enamel wear increased between 4 and 8 years, with significantly higher values for enamel after 6 years (p<0.05). Luting gap wear increased continuously up to 8 years (p<0.05), with no influence of luting composites (p>0.05) and location of teeth (p>0.05). PMID- 17122162 TI - Roles of CLCA and CFTR in electrolyte re-absorption from rat saliva. AB - A molecular basis for Cl- re-absorption has not been well-characterized in salivary ductal cells. Previously, we found strong expression of a rat homologue proposed to be Ca2+-dependent Cl- channels (rCLCA) in the intralobular ducts of the rat submandibular gland. To address the question as to whether rCLCA and cystic fibrosis transmembrane conductance regulator (CFTR) are involved in Cl- re absorption, we evaluated the electrolyte content of saliva from glands pre treated with a small interfering RNA (siRNA). Retrograde injection into a given submandibular duct of an siRNA designed to knock down either rCLCA or CFTR reduced the expression of each of the proteins. rCLCA and CFTR siRNAs significantly increased Cl- concentration in the final saliva during pilocarpine stimulation. These results represent the first in vivo evidence for a physiological significance of rCLCA, along with CFTR, in transepithelial Cl- transport in the ductal system of the rat submandibular gland. PMID- 17122163 TI - Effects of phenylephrine on transplanted submandibular gland. AB - Autotransplantation of the submandibular gland is a potential treatment for severe kerato-conjunctivitis sicca. However, one of the major barriers to this procedure is that secretions from the transplanted gland decrease shortly after the operation, which may lead to obstruction of Wharton's duct, or even to transplantation failure. Using a rabbit model, we investigated whether phenylephrine could improve the secretion from the transplanted gland. We found that phenylephrine treatment significantly reversed the decrease in salivary secretion after transplantation, enhanced the expressions of alpha1A-, alpha1B-, and alpha1D-adrenoceptor mRNA, and ameliorated atrophy of acinar cells. Furthermore, phenylephrine also induced translocation of aquaporin-5 from the cytoplasm to the apical membrane, and increased the levels of phospho-ERK1/2, ERK1/2, phospho-PKCzeta, and PKCzeta in the transplanted gland. These results indicate that phenylephrine treatment moderates structural injury and improves secretory function in the transplanted submandibular gland through promoting alpha1-adrenoceptor expression and post-receptor signal transduction. PMID- 17122164 TI - Duty time of rabbit jaw muscles varies with the number of activity bursts. AB - The relative duration of muscle activity during a specified period (duty time) varies depending on activity level and time of the day. Since both the number and the length of activity bursts contribute to the duty time, it was hypothesized that these variables would show intra-day variations similar to those of the duty time. To test this, we determined duty times, burst numbers, and burst lengths per hour, in relation to multiple activity levels, in a 24-hour period of concurrent radio-telemetric long-term electromyograms of various rabbit jaw muscles. The marked intra-day variation of the burst number resembled that of the duty time in all muscles, and was in contrast to the relatively invariable mean burst length. Furthermore, the duty times were more highly correlated with the number than with the length of bursts at all activity levels. Thus, the variation of the duty time in rabbit jaw muscles is caused mainly by changes in burst numbers. PMID- 17122165 TI - Bilateral TMJ disk displacement induces mandibular retrognathia. AB - Unilateral non-reducing TMJ disk displacement has been shown to retard mandibular growth on the ipsilateral side, with facial asymmetry a sequela. We hypothesized that bilateral affliction would impair mandibular growth bilaterally, generating mandibular retrognathia. Non-reducing TMJ disk displacement was surgically created in 10 growing New Zealand White rabbits. Ten additional rabbits served as a sham-operated control group. Facial growth was followed in serial cephalograms, with tantalum implants, during a period corresponding to childhood and adolescence in man. The results verified that bilateral non-reducing TMJ disk displacement retarded mandibular growth bilaterally, the extent corresponding to mandibular retrognathia in man. Maxillary growth was also retarded, but to a lesser degree. Growth impairment fluctuated over time, the most striking retardation occurring during periods of general growth acceleration. This should be taken into consideration when orthodontic treatment, aimed at stimulating mandibular growth, is initiated in adolescent individuals with non-reducing TMJ disk displacement. PMID- 17122166 TI - Calcium modulates interactions between bacteria and hydroxyapatite. AB - Bacterial adhesion onto hydroxyapatite is known to depend on the surface properties of both the biomaterial and the bacterial strain, but less is known about the influence of the composition of the aqueous medium. Here, the adhesion of Streptococcus mutans and 3 different Lactobacilli on powdered hydroxyapatite was shown to change with Ca2+ concentration. The effect depends on the surface properties of each strain. Adhesion of Lactobacillus fermentum and salivarius (and of Streptococcus mutans at low Ca2+) was enhanced with increasing Ca2+ concentration. Lactobacillus casei was efficiently removed by adhesion on hydroxyapatite, even without Ca2+ addition, and the effect of this ion was only marginal. The results are interpreted in terms of Ca2+-mediated adhesion, and relative to the hydrophobic properties of each strain and the electrical properties of the bacterial and solid surfaces (electrophoretic mobility). PMID- 17122167 TI - Human saliva proteome and transcriptome. AB - This paper tests the hypothesis that salivary proteins and their counterpart mRNAs co-exist in human whole saliva. Global profiling of human saliva proteomes and transcriptomes by mass spectrometry (MS) and expression microarray technologies, respectively, revealed many similarities between saliva proteins and mRNAs. Of the function-known proteins identified in saliva, from 61 to 70% were also found present as mRNA transcripts. For genes not detected at both protein and mRNA levels, we made further efforts to determine if the counterpart is present. Of 19 selected genes detected only at the protein level, the mRNAs of 13 (68%) genes were found in saliva by RT-PCR. In contrast, of many mRNAs detected only by microarrays, their protein products were found in saliva, as reported previously by other investigators. The saliva transcriptome may provide preliminary insights into the boundary of the saliva proteome. PMID- 17122168 TI - Gingivitis susceptibility and its relation to periodontitis in men. AB - The objective of this cross-sectional study was to evaluate whether gingivitis susceptibility is associated with periodontitis. We analyzed data of 462 men in the VA Dental Longitudinal Study aged 47 to 92 years who had never smoked or had quit smoking 5+ years previously. Multiple logistic regression models, with tooth level bleeding on probing at sites with attachment lossT, E349V) in exon 7 of IRF6 in all the affected members of the family. Since the E349V change may disturb the hydrophobic core and affect regulatory activity of IRF6, it is most likely that the mutation is causative for VWS in this family. Fingernail DNA is thus useful for linkage and mutation analyses, since the fingernail can be easily obtained non-invasively, sent through the mail, and stored for a long period. We emphasize here the usefulness of fingernail DNA for the genetic analysis of a disease. PMID- 17122171 TI - Predicting clustered dental implant survival using frailty methods. AB - The purpose of this study was to predict future implant survival using information on risk factors and on the survival status of an individual's existing implant(s). We considered a retrospective cohort study with 677 individuals having 2349 implants placed. We proposed to predict the survival probabilities using the Cox proportional hazards frailty model, with three important risk factors: smoking status, timing of placement, and implant staging. For a non-smoking individual with 2 implants placed, an immediate implant and in one stage, the marginal probability that 1 implant would survive 12 months was 85.8% (95%CI: 77%, 91.7%), and the predicted joint probability of surviving for 12 months was 75.1% (95%CI: 62.1%, 84.7%). If 1 implant was placed earlier and had survived for 12 months, then the second implant had an 87.5% (95%CI: 80.3%, 92.4%) chance of surviving 12 months. Such conditional and joint predictions can assist in clinical decision-making for individuals. PMID- 17122172 TI - Positive illusions in marital relationships: a 13-year longitudinal study. AB - This study examined the long-term consequences of idealization in marriage, using both daily diary and questionnaire data collected from a sample of 168 newlywed couples who participated in a 4-wave, 13-year longitudinal study of marriage. Idealization was operationalized as the tendency for people to perceive their partner as more agreeable than would be expected based on their reports of their partner's agreeable and disagreeable behaviors. Spouses who idealized one another were more in love with each other as newlyweds. Longitudinal analyses suggested that spouses were less likely to suffer declines in love when they idealized one another as newlyweds. Newlywed levels of idealization did not predict divorce. PMID- 17122173 TI - Pursuit of comfort and pursuit of harmony: culture, relationships, and social support seeking. AB - This research examined whether people from collectivistic cultures are less likely to seek social support than are people from individualistic cultures because they are more cautious about potentially disturbing their social network. Study 1 found that Asian Americans from a more collectivistic culture sought social support less and found support seeking to be less effective than European Americans from a more individualistic culture. Study 2 found that European Americans' willingness to seek support was unaffected by relationship priming, whereas Asian Americans were willing to seek support less when the relationship primed was closer to the self. Study 3 replicated the results of Study 2 and found that the tendency to seek support and expect social support to be helpful as related to concerns about relationships. These findings underscore the importance of culturally divergent relationship patterns in understanding social support transactions. PMID- 17122174 TI - Self-conscious emotions and depression: rumination explains why shame but not guilt is maladaptive. AB - Feelings of shame and guilt are factors associated with depression. However, studies simultaneously investigating shame and guilt suggest that only shame has a strong unique effect, although it is not yet clear which psychological processes cause shame and not shame-free guilt to be related to depression. The authors hypothesized that shame, in contrast to guilt, elicits rumination, which then leads to depression. Therefore, in this study we investigated event-related shame and guilt, event-related rumination, and depression among 149 mothers and fathers following family breakup due to marital separation. Data were analyzed using latent variable modeling. The results confirm that shame but not guilt has a strong unique effect on depression. Moreover, the results show that the effect of shame is substantially mediated by rumination. The results are discussed against the background of self-discrepancies and self-esteem. PMID- 17122175 TI - Protecting threatened identity: sticking with the group by emphasizing ingroup heterogeneity. AB - In two studies (Ns=163, 164), the authors tested the prediction that perceptions of group variability can steer and guide the way that loyalty is expressed in times of identity threat. In both studies, participants were classified as lower or higher identifiers on the basis of their scores on a group identification measure, and manipulations involved group variability perceptions (homogeneous ingroup vs. heterogeneous ingroup) and threat to the ingroup. Higher identifiers presented with a homogeneous ingroup perceived more ingroup homogeneity under threat than when there was no threat. In contrast, higher identifiers who perceived the ingroup initially as heterogeneous perceived more ingroup heterogeneity under threat than in no threat conditions. Lower identifiers perceived more ingroup heterogeneity under threat (vs. no threat) irrespective of manipulated group variability perceptions. Discussion focuses on different ways that group loyalty can be expressed in times of identity threat. PMID- 17122176 TI - Play it safe or go for the gold? A terror management perspective on self enhancement and self-protective motives in risky decision making. AB - Terror management theory (TMT) posits that bolstering self-esteem buffers mortality concerns; accordingly, in past research, heightening mortality salience (MS) increases self-enhancement. However, risky self-esteem-relevant decisions often present a choice between enhancing self-esteem by striving for excellence and protecting self-esteem by avoiding potential failure. Which strategy is preferred under MS? Combining TMT with insights from Steele, Spencer, and Lynch's (1993) resource model, the authors hypothesized and found that MS leads high, but not low, self-esteem participants faced with a risky decision to pursue opportunities for excellence despite substantial risk of failure (Studies 1 and 2); in Study 3, using a more impactful decision, this effect was replicated and it was furthermore found that mortality-salient low-self-esteem participants become more risk-averse. Furthermore, in Study 2, a self-affirmation prime, previously shown to reduce MS-induced defenses, eliminated the self-enhancement effect among high-self-esteem participants. Implications for understanding self esteem, TMT, and risky decision making are briefly discussed. PMID- 17122177 TI - The belief in a just world and immanent justice reasoning in adults. AB - Deciding that negative experiences are punishment for prior misdeeds, even when plausible causal links are missing, is immanent justice (IJ) reasoning (Piaget, 1932/1965). Three studies examined a just world theory analysis of IJ reasoning in adults (Lerner, 1980). Studies 1 and 2 varied the valence of a target person's behavior prior to them experiencing an unrelated negative (car accident, Study 1) or positive (lottery win, Study 2) outcome. Participants viewed the outcomes as the result of prior behavior most when they fit deservingness expectations (good person won the lottery, bad person injured in automobile accident), suggesting that just world concerns influenced IJ reasoning. The lottery-winning finding (Study 2) also extends IJ reasoning to positive experiences. A third study found that a manipulation of just world threat in one context (prolonged or ended suffering of an HIV victim) influenced IJ responses in a subsequent unrelated context (automobile accident scenario). PMID- 17122178 TI - Fertility cycle patterns in motives for sexual behavior. AB - Three studies examined the relationship between fertility, as indexed by day in cycle or salivary estradiol levels, and reported motives for sexual intercourse. Data from university and community samples in both cross-sectional and repeated measures designs showed that sex-for-intimacy motives are endorsed significantly less by women who are relatively more fertile or show higher salivary estrogen levels. Sex for enhancement, self-affirmation, and partner approval also declined, although the data were less consistent. The remaining motives (coping and power) showed no consistent patterns across fertility status. Results suggest that sexual motives are rooted partly in woman's biology and are discussed in terms of plausible evolutionary mechanisms that might promote female "choosiness" at midcycle. PMID- 17122179 TI - Perceiving intergroup conflict: from game models to mental templates. AB - This article puts forward a parsimonious framework for studying subjective perceptions of real-life intergroup conflicts. Four studies were conducted to explore how individuals perceive the strategic properties of the Israeli Palestinian conflict. Studies 1 and 2 found theory-driven associations between people's subjective perception of the conflict's structure as a Chicken, Assurance, or Prisoner's Dilemma game and their ingroup/outgroup perceptions, national identification, religiosity, political partisanship, voting behavior, and right-wing authoritarianism. Studies 3 and 4 manipulated the saliency of the needs for cognitive closure and security, respectively, demonstrating that these needs affect people's endorsement of the game models as descriptions of the conflict. PMID- 17122180 TI - Matching achievement contexts with implicit theories to maximize motivation after failure: a congruence model. AB - Previous research has shown that matching person variables with achievement contexts can produce the best motivational outcomes. The current study examines whether this is also true when matching entity and incremental beliefs with the appropriate motivational climate. Participants were led to believe that a personal attribute was fixed (entity belief) or malleable (incremental belief). After thinking that they failed a test that assessed the attribute, participants performed a second (related) task in a context that facilitated the pursuit of either performance or learning goals. Participants were expected to exhibit greater effort on the second task in the congruent conditions (entity belief plus performance goal climate and incremental belief plus learning goal climate) than in the incongruent conditions. These results were obtained, but only for participants who either valued competence on the attribute or had high achievement motivation. Results are discussed in terms of developing strategies for optimizing motivation in achievement settings. PMID- 17122181 TI - Emotional ties that bind: the roles of valence and consistency of group emotion in inferences of cohesiveness and common fate. AB - In three studies, observers based inferences about the cohesiveness and common fate of groups on the emotions expressed by group members. The valence of expressions affected cohesiveness inferences, whereas the consistency of expressions affected inferences of whether members have common fate. These emotion composition effects were stronger than those due to the race or sex composition of the group. Furthermore, the authors show that emotion valence and consistency are differentially involved in judgments about the degree to which the group as a whole was responsible for group performance. Finally, it is demonstrated that valence-cohesiveness effects are mediated by inferences of interpersonal liking and that consistency-common fate effects are mediated by inferences of psychological similarity. These findings have implications for the literature on entitativity and regarding the function of emotions in social contexts. PMID- 17122182 TI - On the waxing and waning of working memory: action orientation moderates the impact of demanding relationship primes on working memory capacity. AB - The present research examined how action- versus state-oriented individuals (Kuhl & Beckmann, 1994) utilize their working memory capacity under varying situational demands. Participants visualized either a demanding or an accepting person, after which their working memory capacity was assessed. Among action-oriented participants, visualizing a demanding person led to greater operation spans (Study 1) and superior memory for intention-related information (Study 2) than visualizing an accepting person. State-oriented participants displayed the opposite pattern, such that visualizing an accepting person led to greater operation spans (Study 1) and superior memory for intentions (Study 2) than visualizing a demanding person. These findings indicate that action versus state orientation moderates the impact of situational demands on working memory capacity. PMID- 17122186 TI - Improving psychometric methods in health education and health behavior research. PMID- 17122188 TI - Are there public health lessons that can be used to help prevent childhood obesity? PMID- 17122189 TI - Downregulation of NADPH oxidase, antioxidant enzymes, and inflammatory markers in the heart of streptozotocin-induced diabetic rats by N-acetyl-L-cysteine. AB - We investigated the effect of N-acetyl-l-cysteine (NAC) on the expression of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, antioxidant enzymes, and inflammatory markers in diabetic rat hearts. Metabolic parameters, free 15 F(2t)-isoprostane level, protein expression of NADPH oxidase, superoxide dismutase (SOD), heme oxygenase (HO-1), interleukin-6 (IL-6), and cyclooxygenase 2 (COX-2) were analyzed in control and streptozotocin-induced diabetic rats treated with or without NAC in drinking water for 8 wk. The cardiac protein expression of p67(phox) and p22(phox) was increased in diabetic rats, accompanied by increased NADPH-dependent superoxide production. As a compensatory response to the increased NADPH oxidase, the protein expression of Cu-Zn-SOD and HO-1 and the total SOD activity were also increased in diabetic rat hearts. Consequently, cardiac free 15-F(2t)-isoprostane, an index of oxidative stress, was increased in diabetic rats, indicating that the production of reactive oxygen species becomes excessive in diabetic rat hearts. Cardiac inflammatory markers IL-6 and COX-2 were also increased in diabetic rats. NAC treatment prevented the increased expression of p22(phox) and translocation of p67(phox) to the membrane in diabetic rat hearts. Subsequently, the levels of cardiac free 15-F(2t) isoprostane, HO-1, Cu-Zn-SOD, total SOD, IL-6, and COX-2 in diabetic rats were decreased by NAC. Consequently, cardiac hypertrophy was attenuated in diabetic rats treated with NAC. The protective effects of NAC on diabetic rat hearts may be attributable to its protection of hearts against oxidative damage induced by the increased NADPH oxidase and to its reduction in cardiac inflammatory mediators IL-6 and COX-2. PMID- 17122190 TI - In vivo left ventricular functional capacity is compromised in cMyBP-C null mice. AB - Cardiac myosin binding protein-C (cMyBP-C) is a thick filament-associated protein that binds tightly to myosin and has a potential role for modulating myocardial contraction. We tested the hypothesis that cMyBP-C 1) contributes to the enhanced in vivo contractile state following beta-adrenergic stimulation and 2) is necessary for myocardial adaptation to chronic increases in afterload. In vivo pressure-volume relations demonstrated that left ventricular (LV) systolic and diastolic function were compromised under basal conditions in cMyBP-C(-/-) compared with WT mice. Moreover, whereas beta-adrenergic treatment significantly improved ejection fraction, peak elastance, and the time to peak elastance in WT mice, these functional indexes remained unchanged in cMyBP-C(-/-) mice. Morphological and functional changes were measured through echocardiography in anesthetized mice following 5 wk of aortic banding. Adaptation to pressure overload was diminished in cMyBP-C(-/-) mice as characterized by a lack of an increase in posterior wall thickness, increased LV diameter, deterioration of fractional shortening, and prolonged isovolumic relaxation time. These results suggest that the absence of cMyBP-C significantly diminishes in vivo LV function and markedly attenuates the increase in LV contractility following beta adrenergic stimulation or adaptation to pressure overload. PMID- 17122191 TI - Cerebral vascular dysfunction in TallyHo mice: a new model of Type II diabetes. AB - The purpose of this study was to characterize vascular responses and to examine mechanisms of vascular dysfunction in TallyHo mice, a new polygenic model of Type II diabetes. Responses of cerebral arterioles and carotid arteries were examined in vivo by using a cranial window and in vitro by using tissue baths, respectively. Dilatation of cerebral arterioles (baseline diameter = 33 +/- 1 micro m) in response to acetylcholine, but not to nitroprusside, was markedly reduced (P < 0.05) in TallyHo mice. Responses of cerebral arterioles to acetylcholine in TallyHo mice were restored to normal with polyethylene glycol superoxide dismutase (100 U/ml; a superoxide scavenger). Responses to acetylcholine were also greatly impaired (P < 0.05) in the carotid arteries from TallyHo mice. Phenylephrine- and serotonin-, but not to KCl- or U46619-, induced contraction was increased two- to fourfold (P < 0.05) in carotid arteries of TallyHo mice. Responses to phenylephrine and serotonin were reduced to similar levels in the presence of Y-27632 (an inhibitor of Rho kinase; 3 micro mol/l). These findings provide the first evidence that vascular dysfunction is present in TallyHo mice and that oxidative stress and enhanced activity of Rho kinase may contribute to altered vascular function in this genetic model of Type II diabetes. PMID- 17122192 TI - Vascular metabolic dissipation in Murray's law. AB - The metabolic dissipation in Murray's minimum energy hypothesis includes only the blood metabolism. The metabolic dissipation of the vascular tree, however, should also include the metabolism of passive and active components of the vessel wall. In this study, we extend the metabolic dissipation to include blood metabolism, as well as passive and active components of the vessel wall. The analysis is extended to the entire vascular arterial tree rather than a single vessel as in Murray's formulation. The calculations are based on experimentally measured morphological data of coronary artery network and the longitudinal distribution of blood pressure along the tree. Whereas the model includes multiple dissipation sources, the total metabolic consumption of a complex vascular tree is found to remain approximately proportional to the cumulative arterial volume of the unit. This implies that the previously described scaling relations for the various morphological features (volume, length, diameter, and flow) remain unchanged under the generalized condition of metabolic requirements of blood and blood vessel wall. PMID- 17122193 TI - Magnetic resonance imaging of progressive cardiomyopathic changes in the db/db mouse. AB - The db/db mouse is a well-established model of diabetes. Previous reports have documented contractile dysfunction (i.e., cardiomyopathy) in these animals, although the extant literature provides limited insights into cardiac structure and function as they change over time. To better elucidate the natural history of cardiomyopathy in db/db mice, we performed cardiac magnetic resonance (CMR) scans on these animals. CMR imaging was conducted with a 4.7-T magnet on female db/db mice and control db/+ littermates at 5, 9, 13, 17, and 22 wk of age. Gated gradient echo sequences were used to obtain cineographic short-axis slices from apex to base. From these images left ventricular (LV) mass (LVM), wall thickness, end-diastolic volume (LVEDV), and ejection fraction (LVEF) were determined. Additionally, cardiac [(18)F]fluorodeoxyglucose ([(18)F]FDG) PET scanning, pressure-volume loops, and real-time quantitative PCR on db/db myocardium were performed. Relative to control, db/db mice developed significant increases in LVM and wall thickness as early as 9 wk of age. LVEDV diverged slightly later, at 13 wk. Interestingly, compared with the baseline level, LVEF in the db/db group did not decrease significantly until 22 wk. Additionally, [(18)F]FDG metabolic imaging showed a 40% decrease in glucose uptake in db/db mice. Furthermore, contractile dysfunction was observed in 15-wk db/db mice undergoing pressure volume loops. Finally, real-time quantitative PCR revealed an age-dependent recapitulation of the fetal gene program, consistent with a myopathic process. In summary, as assessed by CMR, db/db mice develop characteristic structural and functional changes consistent with cardiomyopathy. PMID- 17122194 TI - Spatial heterogeneity of the restitution portrait in rabbit epicardium. AB - Spatial heterogeneity of repolarization can provide a substrate for reentry to occur in myocardium. This heterogeneity may result from spatial differences in action potential duration (APD) restitution. The restitution portrait (RP) measures many aspects of rate-dependent restitution: the dynamic restitution curve (RC), S1-S2 RC, and short-term memory response. We used the RP to characterize epicardial patterns of spatial heterogeneity of restitution that were repeatable across animals. New Zealand White rabbit ventricles were paced from the epicardial apex, midventricle, or base, and optical action potentials were recorded from the same three regions. A perturbed downsweep pacing protocol was applied that measured the RP over a range of cycle lengths from 1,000 to 140 ms. The time constant of short-term memory measured close to the stimulus was dependent on location. In the midventricle the mean time constant was 19.1 +/- 1.1 s, but it was 39% longer at the apex (P < 0.01) and 23% longer at the base (P = 0.03). The S1-S2 RC slope was dependent on pacing site (P = 0.015), with steeper slope when pacing from the apex than from the base. There were no significant repeatable spatial patterns in steady-state APD at all cycle lengths or in dynamic RC slope. These results indicate that transient patterns of epicardial heterogeneity of APD may occur after a change in pacing rate. Thus it may affect cardiac electrical stability at the onset of a tachycardia or during a series of ectopic beats. Differences in restitution with respect to pacing site suggest that vulnerability may be affected by the location of reentry or ectopic foci. PMID- 17122195 TI - Cellular and molecular determinants of altered Ca2+ handling in the failing rabbit heart: primary defects in SR Ca2+ uptake and release mechanisms. AB - Myocytes from the failing myocardium exhibit depressed and prolonged intracellular Ca(2+) concentration ([Ca(2+)](i)) transients that are, in part, responsible for contractile dysfunction and unstable repolarization. To better understand the molecular basis of the aberrant Ca(2+) handling in heart failure (HF), we studied the rabbit pacing tachycardia HF model. Induction of HF was associated with action potential (AP) duration prolongation that was especially pronounced at low stimulation frequencies. L-type calcium channel current (I(Ca,L)) density (-0.964 +/- 0.172 vs. -0.745 +/- 0.128 pA/pF at +10 mV) and Na(+)/Ca(2+) exchanger (NCX) currents (2.1 +/- 0.8 vs. 2.3 +/- 0.8 pA/pF at +30 mV) were not different in myocytes from control and failing hearts. The amplitude of peak [Ca(2+)](i) was depressed (at +10 mV, 0.72 +/- 0.07 and 0.56 +/- 0.04 microM in normal and failing hearts, respectively; P < 0.05), with slowed rates of decay and reduced Ca(2+) spark amplitudes (P < 0.0001) in myocytes isolated from failing vs. control hearts. Inhibition of sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA)2a revealed a greater reliance on NCX to remove cytosolic Ca(2+) in myocytes isolated from failing vs. control hearts (P < 0.05). mRNA levels of the alpha(1C)-subunit, ryanodine receptor (RyR), and NCX were unchanged from controls, while SERCA2a and phospholamban (PLB) were significantly downregulated in failing vs. control hearts (P < 0.05). alpha(1C) protein levels were unchanged, RyR, SERCA2a, and PLB were significantly downregulated (P < 0.05), while NCX protein was significantly upregulated (P < 0.05). These results support a prominent role for the sarcoplasmic reticulum (SR) in the pathogenesis of HF, in which abnormal SR Ca(2+) uptake and release synergistically contribute to the depressed [Ca(2+)](i) and the altered AP profile phenotype. PMID- 17122196 TI - Downregulation of connexin40 and increased prevalence of atrial arrhythmias in transgenic mice with cardiac-restricted overexpression of tumor necrosis factor. AB - Atrial arrhythmias, primarily atrial fibrillation, have been independently associated with structural remodeling and with inflammation. We hypothesized that sustained inflammatory signaling by tumor necrosis factor (TNF) would lead to alterations both in underlying atrial myocardial structure and in atrial electrical conduction. We performed ECG recording, intracardiac electrophysiology studies, epicardial mapping, and connexin immunohistochemical analyses on transgenic mice with targeted overexpression of TNF in the cardiac compartment (MHCsTNF) and on wild-type (WT) control mice (age 8-16 wk). Atrial and ventricular conduction abnormalities were always evident on ECG in MHCsTNF mice, including a shortened atrioventricular interval with a wide QRS duration secondary to junctional rhythm. Supraventricular arrhythmias were observed in five of eight MHCsTNF mice, whereas none of the mice demonstrated ventricular arrhythmias. No arrhythmias were observed in WT mice. Left ventricular conduction velocity during apical pacing was similar between the two mouse groups. Connexin40 was significantly downregulated in MHCsTNF mice. In contrast, connexin43 density was not significantly altered in MHCsTNF mice, but rather dispersed away from the intercalated disks. In conclusion, sustained inflammatory signaling contributed to atrial structural remodeling and downregulation of connexin40 that was associated with an increased prevalence of atrial arrhythmias. PMID- 17122197 TI - Cardiac effects of postconditioning depend critically on the duration of index ischemia. AB - Postconditioning (POC) is known as the phenomenon whereby brief intermittent ischemia applied at the onset of reperfusion following index ischemia limits myocardial infarct size. Whereas there is evidence that the algorithm of the POC stimulus is an important determinant of the protective efficacy, the importance of the duration of index ischemia on the outcome of the effects of POC has received little attention. Pentobarbital sodium-anesthetized Wistar rats were therefore subjected to index ischemia produced by coronary artery occlusions (CAO) of varying duration (15-120 min) followed by reperfusion, without or with postconditioning produced by three cycles of 30-s reperfusion and reocclusion (3POC30). 3POC30 limited infarct size produced by 45-min CAO (CAO45) from 45 +/- 3% to 31 +/- 5%, and CAO60 from 60 +/- 3% to 47 +/- 6% (both P < or = 0.05). In contrast, 3POC30 increased infarct size produced by CAO15 from 3 +/- 1% to 19 +/- 6% and CAO30 from 36 +/- 6 to 48 +/- 4% (both P < or = 0.05). This deleterious effect of 3POC30 was not stimulus sensitive because postconditioning with 3POC5 and 3POC15 after CAO30 also increased infarct size. The cardioprotection by 3POC30 after CAO60 was accompanied by an increased stimulation of Akt phosphorylation at 7 min of reperfusion and a 36% lower superoxide production, measured by dihydroethidium fluorescence, after 2 h of reperfusion. Consistent with these results, cardioprotection by 3POC30 was abolished by phosphatidylinositol-3-OH-kinase inhibition, as well as nitric oxide (NO) synthase inhibition. The deleterious effect of 3POC30 after CAO15 was accompanied by an increased superoxide production with no change in Akt phosphorylation and was not affected by NO synthase inhibition. In conclusion, the effect of cardiac POC depends critically on the duration of the index ischemia and can be either beneficial or detrimental. These paradoxical effects of POC may be related to the divergent effects on Akt phosphorylation and superoxide production. PMID- 17122200 TI - You get what you pay for. PMID- 17122201 TI - Case scheduling for dummies. PMID- 17122202 TI - Pro: Aprotinin has a good efficacy and safety profile relative to other alternatives for prevention of bleeding in cardiac surgery. PMID- 17122203 TI - Con: Aprotinin has a good efficacy and safety profile relative to other alternatives for prevention of bleeding in cardiac surgery. PMID- 17122204 TI - Pro: Heparin-coated circuits should be used for cardiopulmonary bypass. PMID- 17122205 TI - Con: Heparin-bonded cardiopulmonary bypass circuits should be routine for all cardiac surgical procedures. PMID- 17122206 TI - Temperature during cardiopulmonary bypass: the discrepancies between monitored sites. AB - We performed studies in patients to determine whether temperature recordings from sites commonly monitored during hypothermic cardiopulmonary bypass adequately reflect cerebral temperature. In Study I (n = 12), temperatures monitored in the jugular bulb (JB) were compared with those recorded in the nasopharynx, esophagus, bladder, and rectum. In Study II (n = 30), temperature was also monitored in the arterial outlet of the membrane oxygenator. A calibrated recorder continuously and simultaneously recorded all temperatures. Study I found large temperature discrepancies between the JB and all other body sites during cooling and rewarming. There was considerable interindividual variability in the degree of discrepancy between the JB and other sites. Study II produced similar results but also showed that JB temperature reached equilibration with the temperature of blood entering the patient via the arterial outlet of the membrane oxygenator after cooling for 3.3 +/- 1.3 min and after rewarming for 16.5 +/- 5.5 min. Analysis of variance revealed that this arterial outlet site had the smallest average discrepancy of all temperature sites relative to the JB site (P < 0.001). In summary, temperatures measured in body sites over-estimated JB temperature during cooling and under-estimated it during rewarming, whereas arterial outlet blood temperature provided a good approximation. PMID- 17122207 TI - A comparison of bilateral with single internal mammary artery grafts on postoperative mediastinal drainage and transfusion requirement. AB - The superiority of the left internal mammary artery (LIMA) graft over autogenous saphenous vein as a bypass conduit in coronary artery bypass surgery has been well established. Early and late patency rates of bilateral internal mammary artery (BIMA) grafts exceed those of vein grafts, and patients who receive BIMA have improved long-term survival rates and more freedom from reoperations and other cardiac events. But because of other concerns, particularly the question of increased risk of postoperative bleeding, controversy still surrounds the perioperative period. In the present study we sought to determine whether BIMA grafting was an independent risk factor of postoperative bleeding and of blood product use in patients undergoing primary elective coronary artery revascularization. For this purpose, 33 consecutive patients scheduled for BIMA grafting were matched with 66 patients operated on by single LIMA grafting. Patients in the LIMA group had significantly less postoperative mediastinal drainage than those in the BIMA group (median: 722 vs 920 mL, P = 0.0001). Fifty six patients received blood products (56% vs 51% in LIMA and BIMA groups, respectively; P = 0.67). In multivariate analysis, BIMA and operative duration were independent predictors of increased postoperative drainage. Nevertheless, in logistic regression, BIMA was not significantly associated with blood product use, unlike precardiopulmonary bypass hematocrit and duration of surgery (OR and 95% CI: 0.89 [0.80-0.96] P = 0.01; 1.009 [1.001-1.019] P = 0.04, for an increase of 1% in hematocrit and 1 min in duration of surgery, respectively). In conclusion, these data support the idea that BIMA graft slightly increases postoperative drainage but not transfusion requirement. PMID- 17122208 TI - Perioperative amino acid infusion improves recovery and shortens the duration of hospitalization after off-pump coronary artery bypass grafting. AB - Perioperative amino acid infusion helps maintain core temperature and improves patient outcomes after gynecologic and orthopedic surgery. In the present study we prospectively determined the effect of amino acid infusion on esophageal core temperature and postoperative outcomes during off-pump coronary artery bypass grafting (CABG). One-hundred-eighty consecutive patients undergoing primary elective or urgent off-pump CABG were randomly divided into two groups: the i.v. amino acid infusion group (4 kJ kg(-1) h(-1) starting 2 h before surgery) and the saline infusion group (similar period and volume of saline infusion). The esophageal core temperature at the end of surgery was 35.6 (35.3-35.8) degrees C [mean (95% confidence interval)] in the saline infusion group and 36.1 degrees C (35.9-36.3) degrees C in the amino acid infusion group (P = 0.01). Kaplan-Meier analysis demonstrated that patients given amino acids required a significantly shorter duration of postoperative mechanical ventilation than patients given saline [median (95% confidence interval), 3.0 (2.5-3.9) vs 4.5 (3.8-5.8) h; P = 0.01]. Furthermore, intensive care unit stay [20 (19.5-38.4) vs 44 (21-45) h; P = 0.001] and days until fit for discharge from hospital [10 (9-11) vs 12 (11-13) days; P = 0.004] were significantly shorter in patients given amino acid. Perioperative amino acid infusion in patients undergoing off-pump CABG effectively minimizes intraoperative hypothermia and improves postoperative recovery. PMID- 17122209 TI - Activation of a neutrophil-derived inflammatory response in the airways during cardiopulmonary bypass. AB - Cardiopulmonary bypass (CPB) is believed to cause postoperative lung dysfunction. To more closely examine the inflammatory processes occurring in the airways during CPB, we serially measured inflammatory mediators, with the assistance of a new bronchoscopic microsample probe, in 11 patients undergoing repair of aortic arch aneurysms. Epithelial lining fluid (ELF) and arterial blood were sampled simultaneously after induction of anesthesia, at the time of pulmonary reperfusion, and at the end of surgery. A decrease in the PaO2/FiO2 ratio was observed at the end of surgery (P = 0.029). Although the ELF concentrations of interleukin (IL)-8, IL-6, and neutrophil elastase had increased significantly at the end of surgery (median = 23,200, 1818, and 12,900 microg/mL, respectively), they did not correlate with the degree of hypoxemia. Neutrophil elastase increased significantly at the time of pulmonary reperfusion, before IL-8 and IL 6, and independently of blood transfusions. At the end of surgery, IL-6 in ELF correlated with total blood transfusion volume (rho = 0.731, P = 0.011). These results indicate that a neutrophil-derived inflammatory response is activated in the airway in the early phase of CPB. PMID- 17122210 TI - Inhibition of apoptotic protein p53 lowers the threshold of isoflurane-induced cardioprotection during early reperfusion in rabbits. AB - INTRODUCTION: Exposure to isoflurane before and during early reperfusion protects against myocardial infarction by activating phosphatidylinositol-3-kinase (PI3K) mediated signaling. The apoptotic protein, p53, is regulated by PI3K, and inhibition of p53 protects against ischemic injury. We tested the hypothesis that p53 inhibition lowers the threshold of isoflurane-induced postconditioning in vivo. METHODS: Rabbits (n = 73) instrumented for hemodynamic measurement and subjected to a 30-min left anterior descending coronary artery occlusion and 3-h reperfusion received 0.9% saline (control), isoflurane (0.5 or 1.0 minimum alveolar concentration [MAC]) administered for 3 min before and 2 min after reperfusion, the p53 inhibitor pifithrin-alpha (1.5 or 3.0 mg/kg), or 0.5 MAC isoflurane plus 1.5 mg/kg pifithrin-alpha. Other rabbits received 3.0 mg/kg pifithrin-alpha or 0.5 MAC isoflurane plus 1.5 mg/kg pifithrin-alpha after pretreatment with the selective PI3K inhibitor wortmannin (0.6 mg/kg) or the mitochondrial permeability transition pore opener atractyloside (5 mg/kg). RESULTS: Isoflurane (1.0 but not 0.5 MAC), pifithrin-alpha (3.0 but not 1.5 mg/kg), and the combination of 0.5 MAC isoflurane plus 1.5 mg/kg pifithrin-alpha significantly (P < 0.05) reduced infarct size (21% +/- 4%, 43% +/- 7%, 22% +/- 4%, 45% +/- 4%, and 28% +/- 3% [mean +/- sd], respectively, of left ventricular area at risk; triphenyltetrazolium chloride staining) when compared with control (45% +/- 2%). Atractyloside, but not wortmannin, abolished 3.0 mg/kg pifithrin alpha-induced cardioprotection, whereas atractyloside and wortmannin blocked reductions in infarct size produced by 0.5 MAC isoflurane plus 1.5 mg/kg pifithrin-alpha. CONCLUSION: The results indicate that inhibition of the apoptotic protein p53 lowers the threshold of isoflurane-induced cardioprotection during early reperfusion in vivo. PMID- 17122211 TI - Dynamic left ventricular inflow obstruction associated with a left atrial myxoma. PMID- 17122212 TI - Acute high-output failure from an aortoventricular fistula due to a ruptured sinus of Valsalva aneurysm after blunt chest trauma. PMID- 17122213 TI - Tricuspid regurgitation caused by eustachian valve endocarditis. PMID- 17122214 TI - Placement of the TandemHeart percutaneous left ventricular assist device. PMID- 17122215 TI - Transesophageal echocardiography of the quadricuspid aortic valve. PMID- 17122216 TI - ASE/SCA recommendations and guidelines for continuous quality improvement in perioperative echocardiography. PMID- 17122217 TI - Scheduling elective pediatric procedures that require anesthesia: the perspective of parents. AB - Daily variability in volume of elective pediatric procedures that require anesthesia may lead to an imbalance between available operating room resources and case load. Longer intervals between scheduling and the surgical date generally result in higher operating room utilization. In this study, we sought to determine which factors influence when parents schedule their children for procedures. We also aimed to identify parents' ideal and longest acceptable waiting intervals and determine whether type of procedure, for example, affects scheduling. From a convenience sample of 250 randomly selected parents of children presenting for elective surgery, 236 completed surveys were analyzed. The remaining 14 surveys were not returned. Overall, parents scheduled their child's procedure a median of 4.3 wk (interquartile range 2.0-8.6) in advance and reported an ideal waiting interval of 3 wk (interquartile range 2-4), and longest acceptable interval of 6 wk (interquartile range 4-10). Parents were willing to wait longer to schedule cardiac (4 wk, P = 0.004) and plastic (3.5 wk, P = 0.024) surgery when compared with general surgical procedures. Overall, parents ranked severity of the child's illness, earliest available time, and surgeon's suggested date as the three most important factors influencing when their child's surgery is scheduled. The timetable for scheduling procedures was highly correlated with both mother and father having available time off work (tau(b) = 0.72, P < 0.0001). Surprisingly, parents did not show a preference for scheduling cases during vacation or summer months. PMID- 17122218 TI - Anesthetic management of a young adult with complex congenital heart disease and bronchopleural fistula for rigid bronchoscopy. AB - The number of adults with congenital heart disease and those who require anesthetic care are increasing. We describe the anesthetic management of a young adult with palliated complex congenital heart disease and a chronic postsurgical bronchopleural fistula for rigid bronchoscopy. Perioperative considerations in the care of patients with single ventricle physiology for noncardiac procedures are reviewed. Specific requirements for rigid bronchoscopy are discussed in addition to the anesthetic implications of a bronchopleural fistula and particular concerns in the patient with single ventricle physiology. PMID- 17122219 TI - Outpatient management of continuous peripheral nerve catheters placed using ultrasound guidance: an experience in 620 patients. AB - BACKGROUND: Continuous peripheral nerve block (CPNB) is an optimal choice for analgesia after orthopedic procedures, but is not commonly used in outpatients because of concern regarding the possibility of catheter-related complications. In addition, it may be difficult to provide adequate patient access to physicians in this setting. We present 620 outpatients who were treated with CPNB using an established protocol. METHODS: All catheters were placed using direct ultrasound visualization. These patients received extensive oral and written preoperative instruction and were provided continuous telephone access to the anesthesiologist during the postoperative period. All patients were also contacted at home by telephone on the first postoperative day. In addition, each patient was seen and examined by the surgeon within 2 wk of hospital discharge. RESULTS: Of the 620 patients, there were 190 interscalene (brachial plexus), 206 fascia iliaca (femoral nerve), and 224 popliteal fossa (sciatic nerve) catheters. Two patients (0.3%) had complications related to the nerve block. In both of these patients, the symptoms resolved within 6 wk of surgery. Twenty-six patients (4.2%) required postoperative interventions by the anesthesiologist. One patient returned to the hospital for catheter removal. CONCLUSIONS: In this large series of outpatients treated with CPNB, there were surprisingly few interventions requiring an anesthesiologist. Likewise, patients were able to manage and remove their catheters at home without additional follow-up. This suggests that with adequate instruction and telephone access to health care providers, patients are comfortable with managing and removing CPNB catheters at home. PMID- 17122220 TI - Small-dose ketamine reduces the pain of propofol injection. AB - BACKGROUND: I.v. injection of propofol during anesthetic induction induces pain. Ketamine has been shown to reduce injection pain. In this study, we established the optimal dose of ketamine to prevent the pain of injection with propofol. METHODS: Two hundred forty patients presenting for elective surgery were randomly allocated into eight groups; five groups during the first part of the study and three groups during the second part. In Part 1, patients received saline (Group S), lidocaine (Group L), ketamine 10 microg/kg (Group K10), 50 microg/kg (Group K50), or 100 microg/kg (Group K100), respectively, immediately followed by propofol 2.5 mg/kg. In Part 2, the optimal dose of ketamine (100 microg/kg) was administered 3 min before propofol (Group Pre), mixed with propofol solution (Group KP), or after oral midazolam premedication (Group M). An anesthesiologist blinded to the study group monitored each patient's pain score at 5-s intervals. RESULTS: In Part 1, the incidence and intensity of pain were the lowest in the K100 and L groups (P < 0.001). In Part 2, the patients in the K100 and M groups had significantly lower pain scores compared with the KP and Pre groups (P < 0.05). During induction, there were no significant intergroup differences in mean arterial blood pressure and heart rate in all groups. CONCLUSIONS: Administration of ketamine 100 microg/kg immediately before propofol injection provided the optimal dose and timing to reduce propofol-induced pain on injection. PMID- 17122221 TI - The effects of melatonin premedication on propofol and thiopental induction dose response curves: a prospective, randomized, double-blind study. AB - BACKGROUND: The effect of melatonin on the intraoperative requirements for i.v.anesthetics has not been documented. We studied the effect of melatonin premedication on the propofol and thiopental dose-response curves for abolition of responses to verbal commands and eyelash stimulation. METHODS: This prospective, randomized, double-blind study included 200 adults with ASA physical status I. Patients received either 0.2 mg/kg melatonin or a placebo orally for premedication (n = 100 per group). Approximately 50 min later, subgroups of 10 melatonin and 10 placebo patients were administered various doses of propofol (0.5, 1.0, 1.5, 2.0, or 2.4 mg/kg) or thiopental (2.0, 3.0, 4.0, 5.0, or 6.0 mg/kg) for anesthetic induction. The ability of each patient to respond to the command, "open your eyes," and the disappearance of the eyelash reflex were assessed 60 s after the end of the injection of propofol or thiopental. Dose response curves were determined by probit analysis. RESULTS: Melatonin premedication decreased thiopental ED50 values for loss of response to verbal command and eyelash reflex from 3.4 mg/kg (95% confidence interval, 3.2-3.5 mg/kg) and 3.7 mg/kg (3.5-3.9 mg/kg) to 2.7 mg/kg (2.6-2.9 mg/kg) and 2.6 mg/kg (2.5-2.7 mg/kg), respectively (P < 0.05). Corresponding propofol ED50 values decreased from 1.5 mg/kg (1.4-1.6 mg/kg) and 1.6 mg/kg (1.5-1.7 mg/kg) to 0.9 mg/kg (0.8-0.96 mg/kg) and 0.9 mg/kg (0.8-0.95 mg/kg), respectively (P < 0.05). CONCLUSIONS: Melatonin premedication significantly decreased the doses of both propofol and thiopental required to induce anesthesia. PMID- 17122222 TI - Isoflurane and propofol have similar effects on spinal neuronal windup at concentrations that block movement. AB - BACKGROUND: We investigated the actions of isoflurane and propofol on neuronal windup in the spinal cord of intact rats. We hypothesized that propofol would depress windup more than isoflurane. METHODS: In a cross-over design, rats received 0.8 and 1.2 minimum alveolar concentration (MAC) isoflurane and 0.8 and 1.2 ED50 (effective dose(50%)) of propofol, as recordings were made from single units in the lumbar cord (n = 13). Electrical stimuli were applied (20 stimuli at 0.1, 1, and 3 Hz). Neuronal responses were analyzed for those occurring in the C fiber range (100-400 ms after each stimulus), combined C-fiber and afterdischarge range (100-1000 ms) and the 100-333 ms range for the 3 Hz stimuli. Absolute windup was also calculated (the sum of action potentials for 20 stimuli - 20 x response to the first stimulus). RESULTS: At 1 Hz, total action potentials (mean, standard error) summed across the 20 stimuli (100-1000 ms range) were 571 +/- 106 and 742 +/- 214 for isoflurane (at 0.8 and 1.2 MAC) and 586 +/- 148 and 641 +/- 143 for propofol (at 0.8 and 1.2 ED50), respectively (P = NS); corresponding values for the 0.1 Hz stimuli were 345 +/- 104, 370 +/- 108, 430 +/- 86, and 403 +/- 106 (P = NS), and for the 3 Hz stimuli (100-333 ms range) were 266 +/- 66, 333 +/- 76, 343 +/- 85, and 252 +/- 72 (P = NS). Absolute windup in the 100-1000 ms range was greater for 1.2 MAC isoflurane at 1 Hz (445 +/- 82, P < 0.01), when compared with absolute windup at 0.8 MAC isoflurane and 0.8 and 1.2 ED50 propofol (232 +/- 31, 88 +/- 65, and 210 +/- 41, respectively). CONCLUSIONS: These data suggest that isoflurane and propofol have similar effects on neuronal windup in the spinal cord, although there was enhanced absolute windup at 1.2 MAC isoflurane for the 1 Hz stimulus. PMID- 17122223 TI - The differential effects of nitrous oxide and xenon on extracellular dopamine levels in the rat nucleus accumbens: a microdialysis study. AB - Dopamine release in the nucleus accumbens (NAC) plays a crucial role in the action of various psychotropic and addictive drugs, such as antagonists of the N methyl-D-aspartate subtype of the glutamate. Although both nitrous oxide and xenon are N-methyl-D-aspartate receptor antagonists, they differ in their potential for producing neuropsychological toxicity; therefore, we decided to examine their effects on both spontaneous and ketamine-induced extracellular dopamine levels in the NAC. A microdialysis probe was implanted into the NAC in each of 35 rats, which were randomly assigned to one of six groups: exposure to 40% O2, exposure to 60% nitrous oxide (0.27 MAC), exposure to 43% xenon (0.27 MAC) for 60 min, and three groups exposed to either 40% O2, 60% nitrous oxide, or 43% xenon for 70 min and 80 mg/kg ketamine was given i.p. 10 min after the initiation of gas exposure. Perfusate samples were collected every 20 min, and the dopamine levels were measured using a high-performance liquid chromatography system. Nitrous oxide, but not xenon, significantly increased the dopamine level. Ketamine significantly increased the dopamine level, and this was significantly inhibited by xenon, but not by nitrous oxide. These data suggest that the difference in neuropsychological activity between nitrous oxide and xenon is partly due to their differential effects on the mesolimbic dopamine system. PMID- 17122224 TI - The effect of too much intravenous lidocaine on bispectral index. AB - Systemic local anesthetics have beneficial perioperative properties and have an anesthetic-sparing effect. To assess depth of anesthesia during lidocaine infusion, it would be important to know the effect of systemic local anesthetics on bispectral index (BIS). However, this has not been investigated. We report an inadvertent overdose of i.v. lidocaine in a patient monitored with BIS. BIS decreased to 0 for approximately 15 min, indicating that lidocaine and sevoflurane interact to decrease BIS. PMID- 17122225 TI - Serotonin syndrome from the interaction of cyclobenzaprine with other serotoninergic drugs. AB - Serotonin syndrome is a potentially lethal adverse drug reaction that may occur in patients taking proserotoninergic medications. Drug interactions are often responsible for the causation of this syndrome. We report two cases of severe serotonin syndrome induced by the administration of cyclobenzaprine in postoperative patients already receiving another proserotoninergic drug (phenelzine in one case and duloxetine in the other). In both cases, symptoms of autonomic instability and severe agitation started within hours of initiation of cyclobenzaprine and fully resolved within 3 days after discontinuing the proserotoninergic drugs. We conclude that cyclobenzaprine should be used with extreme caution in patients receiving other serotonin-enhancing drugs; these patients should be closely monitored for manifestations of serotonin syndrome. PMID- 17122226 TI - Does monitoring bispectral index or spectral entropy reduce sevoflurane use? AB - A decrease in volatile anesthetic consumption has been demonstrated using bispectral index (BIS), whereas data concerning spectral entropy are lacking. One hundred and forty adult patients scheduled for surgical procedures lasting more than 1 h were prospectively randomized to receive an anesthetic controlled either by BIS or by spectral entropy or solely by clinical variables. Anesthesia was induced with propofol and sufentanil. Sufentanil was infused continuously thereafter. Sevoflurane was administered in 1 L/min O2/N2O. The sevoflurane concentration was adjusted according to conventional clinical variables in the standard practice group, whereas the 40-60 interval was applied for the BIS and spectral entropy-guided groups. The sevoflurane vaporizer was weighed before and after anesthesia, and consumption was calculated. Groups were comparable for demographic data except for weight (heavier in the spectral entropy-guided group, P < 0.05). Compared with standard practice, patients with BIS or spectral entropy monitoring required 29% less sevoflurane (normalized sevoflurane consumption to the weights of the patients and to the durations of anesthesia; both P < 0.03) and a similar sufentanil dose. An unintended improvement in the standard practice group (positive bias) was observed. In conclusion, BIS and spectral entropy monitoring have the same sparing effect of sevoflurane. PMID- 17122227 TI - Arterial versus plethysmographic dynamic indices to test responsiveness for testing fluid administration in hypotensive patients: a clinical trial. AB - In the present study, we compared indices of respiratory-induced variation obtained from direct arterial blood pressure measurement with analogous indices obtained from the plethysmogram measured by the pulse oximeter to assess the value of these indices for predicting the cardiac output increase in response to a fluid challenge. Thirty-two fluid challenges were performed in 22 hypotensive patients who were also monitored with a pulmonary artery catheter. Hemodynamic and plethysmographic data were collected before and after intravascular volume expansion. Patients were classified as nonresponders if their cardiac index did not increase by 15% from baseline. Nonresponding patients had both lower arterial pulse variation ([10 +/- 4]% vs [19 +/- 13]%, P = 0.020) and lower plethysmographic pulse variation ([12 +/- 7]% vs [21 +/- 14]%, P = 0.034) when compared with responders. Fluid responsiveness was similarly predicted by arterial and plethysmographic pulse variations (area under ROC curve 0.74 vs 0.72, respectively, P = 0.90) and by arterial and plethysmographic systolic variation (area under ROC curve 0.64 vs 0.72, respectively, P = 0.50). Nonresponders were identified by changes in pulse variation both on arterial and plethysmographic waveform (area under ROC curve 0.80 vs 0.87, respectively, P = 0.40) and by changes in arterial and plethysmographic systolic variations (area under ROC curve 0.84 vs 0.80, respectively, P = 0.76). In the population studied, plethysmographic dynamic indices of respiratory-induced variation were just as useful for predicting fluid responsiveness as the analogous indices derived from direct arterial blood pressure measurement. These plethysmographic indices could provide a noninvasive tool for predicting the cardiac output increase by administering fluid. PMID- 17122228 TI - Automated external defibrillators do not recommend false positive shocks under the influence of electromagnetic fields present at public locations. AB - Electromagnetic fields (EMF) reduce the signal quality of electrocardiograms and may lead to the misinterpretation by automated external defibrillators (AED). We designed this investigation as a prospective study, with a randomized sequence of AED applications on healthy volunteers. We chose busy public places where public access defibrillation was possible as test locations. Strong EMF were sought and found at train stations next to accelerating and decelerating trains. The primary outcome variable was the absolute number of shocks advised in the presence of sinus rhythm by five commonly used AED in Austria. For data analysis, the statistician was blinded in regard to the AED models tested. Data analysis was based on a per protocol evaluation. Of 390 tests run, 0 cases of false positive results occurred (95% CI: 0-0.77). AED can be regarded as safe, even with the interference of EMF present at train stations. PMID- 17122229 TI - Acoustic monitoring of double-lumen ventilated lungs for the detection of selective unilateral lung ventilation. AB - One-lung intubation (OLI) is among the most common complications of endotracheal intubation. None of the monitoring tools now available has proved effective for its early detection. In this study we investigated the efficacy of acoustic analysis for the detection of OLI. We collected lung sounds from 11 patients undergoing thoracic surgery requiring the placement of a double-lumen tube. Recordings of separate lung ventilation were performed after induction and confirmation of adequate tube positioning, before surgery. Samples of lung sounds were collected by three piezoelectric microphones, one on each side of the chest and one on the right forearm, for background noise sampling. The samples were filtered, the signals' energy envelopes were calculated, and segmentation to breath and rest periods was performed. Each respiration was classified into one of the three categories: bilateral ventilation, selective right-lung ventilation, or selective left-lung ventilation, on the basis of the ratio between the energy signals of each lung. OLI was accurately identified in 10 of the 11 patients during right OLI and in all 11 patients during left OLI. This study suggests that acoustic monitoring is effective for the detection of selective lung ventilation and may be useful for early diagnosis of OLI. PMID- 17122230 TI - Holiday and weekend operating room on-call staffing requirements. AB - BACKGROUND: Every facility that performs cases on holidays has in some way decided on its operating room (OR) and anesthesia staffing for holidays. Previous studies have not examined how best to calculate appropriate holiday staffing. METHODS: We analyzed weekend and holiday data from a university hospital. RESULTS: There were high rank correlations between the number of cases starting during each 12-h period of a holiday, the total hours of OR time used, and the patients and surgeons waiting for cases to start. Weekend and holiday 12-h periods were divided into 8 categories (e.g., Saturday 7:00 am to 7:00 pm). There was perfect rank correlation between the mean number of cases starting during each 12-h period and appropriate staffing during the 12-h period, whether quantified by total hours of cases or by the under-utilized and over-utilized OR time resulting from staffing decisions. CONCLUSIONS: The number of cases starting during each period of a holiday is a statistically valid end point for OR managers to use to evaluate how busy holidays are relative to weekend days. To be useful, the statistic must be combined with mathematically valid assessments of appropriate weekend staffing on-call, whether in-house, or from home. PMID- 17122231 TI - The impact of service-specific staffing, case scheduling, turnovers, and first case starts on anesthesia group and operating room productivity: a tutorial using data from an Australian hospital. AB - BACKGROUND: In this tutorial, we consider the impact of operating room (OR) management on anesthesia group and OR labor productivity and costs. Most of the tutorial focuses on the steps required for each facility to refine its OR allocations using its own data collected during patient care. METHODS: Data from a hospital in Australia are used throughout to illustrate the methods. OR allocation is a two-stage process. During the initial tactical stage of allocating OR time, OR capacity ("block time") is adjusted. For operational decision-making on a shorter-term basis, the existing workload can be considered fixed. Staffing is matched to that workload based on maximizing the efficiency of use of OR time. RESULTS: Scheduling cases and making decisions on the day of surgery to increase OR efficiency are worthwhile interventions to increase anesthesia group productivity. However, by far, the most important step is the appropriate refinement of OR allocations (i.e., planning service-specific staffing) 2-3 mo before the day of surgery. CONCLUSIONS: Reducing surgical and/or turnover times and delays in first-case-of-the-day starts generally provides small reductions in OR labor costs. Results vary widely because they are highly sensitive both to the OR allocations (i.e., staffing) and to the appropriateness of those OR allocations. PMID- 17122232 TI - A system and process redesign to improve perioperative antibiotic administration. AB - Surgical infection is a leading cause of patient injury, mortality, and excess health care costs. As part of a collaborative effort, we instituted three main focuses for perioperative antibiotic administration: appropriate selection of antibiotics, administration of antibiotics within 60 min before incision, and discontinuation of prophylactic antibiotics within 24 h of surgery. Anesthesiologists were identified as the practitioners most likely to accomplish the successful administration of antibiotics within 60 min before incision. Changes were made in ordering, documentation, and antibiotic preparation. Education was provided to all operating room staff at meetings and grand round presentations. Results were prominently displayed, and feedback was provided. The baseline appropriate antibiotic selection was 82% and is now 95%. The preintervention administration-incision time was 79 (range, 32-380) min, with 11% within the 60 min before incision. The administration-incision time is currently 19 (range, 0-95) min, and the number within 60 min is 95%. Before the institution of the process, the rate of surgical site infections was 3.8%, and is now approximately 1.4%. We describe our process used to improve antibiotic administration. During this time, the surgical site infection rate has been significantly reduced. PMID- 17122233 TI - Changes in the serum proteome of patients with sepsis and septic shock. AB - BACKGROUND: Sepsis is still the leading cause of death in the intensive care unit. Our goal was to elucidate potential early differences in serum between survivors (SURV) and non-survivors (NON-SURV) on day 28. METHODS: We applied proteomic technology to serum samples of patients with sepsis and septic shock. Serum samples from 18 patients with sepsis and septic shock were obtained during the first 12 h after diagnosis of septic shock. Patients were grouped into SURV and NON-SURV on day 28. RESULTS: Seven patients survived and 11 patients died. Using proteome analysis, two-dimensional gel electrophoresis detected more than 200 spots per gel. A differential protein expression was discovered between SURV and NON-SURV, whereby protein alterations not yet described in sepsis were revealed. CONCLUSIONS: Our results show that proteomic profiling is a useful approach for detecting protein expression dynamics in septic patients, and may bring us closer to achieving a comprehensive molecular profiling compared with genetic studies alone. PMID- 17122234 TI - The effect of electroencephalogram-targeted high- and low-dose propofol infusion on histopathological damage after traumatic brain injury in the rat. AB - BACKGROUND: Propofol is commonly used to sedate patients after traumatic brain injury. However, the dose-dependent neuroprotective effects of propofol after head trauma are unknown. We compared histopathological damage after 6 h of electroencephalogram-targeted high- and low-dose propofol infusion in rats subjected to controlled cortical impact (CCI). METHODS: Animals were randomly assigned to CCI/propofol with electroencephalogram burst-suppression-ratio 1%-5% (CCI/lowprop), CCI/propofol with burst-suppression-ratio 30%-40% (CCI/highprop), control group CCI/1.0 vol % halothane (CCI/halo), or sham group with halothane anesthesia (SHAM/halo). Brain slices were stained with kresyl violet (KV) and hematoxylin/eosin (HE) to evaluate lesion volume, number of eosinophilic cells, and activation of caspase-3 in the hippocampus. RESULTS: Lesion volume (mm3) and number of eosinophilic cells in the hippocampus did not differ significantly [lesion volumes: CCI/lowprop 31.55 +/- 14.66 (KV) and 53.77 +/- 8.62 (HE); CCI/highprop 33.81 +/- 10.57 (KV) and 52.30 +/- 11.55 (HE); CCI/halo 36.42 +/- 17.06 (KV) and 57.95 +/- 8.49 (HE)]. Activation of caspase-3 occurred in the ipsilateral hippocampus in all CCI-groups. CONCLUSION: Despite different levels of cortical neuronal function, there were no relevant differences in the short term histopathological damage. These results challenge the view that the neuroprotective effect of propofol relates to the suppression of cerebral metabolic demand. PMID- 17122235 TI - The effect of a lidocaine test dose on analgesia and mobility after an epidural combination of neostigmine and sufentanil in early labor. AB - We previously demonstrated the effectiveness of epidural sufentanil and the cholinesterase inhibitor, neostigmine, to initiate selective labor analgesia. Because the traditional lidocaine plus epinephrine test dose (TD) may alter the effect of subsequent epidural drugs, we undertook this investigation to evaluate the impact of a lidocaine TD on analgesia from a combination of epidural neostigmine plus sufentanil administered in early labor. Eighty healthy parturients were randomly allocated to two groups to receive a 3 mL-TD, either lidocaine 2%-epinephrine (1:200,000) or saline-epinephrine (1:200,000), followed 3 min later by epidural neostigmine 500 microg plus sufentanil 10 microg. Pain scores were recorded for 30 min after injection, as was the time elapsed from initial bolus until request for supplemental analgesia. Thirty minutes after injection, adequacy of motor function was evaluated by the parturient's ability to sit, stand up, bend her knees, and walk. Lidocaine TD hastened the onset (5 min vs 15 min) and increased duration (122 +/- 53 min vs 98 +/- 54 min; P = 0.02) of analgesia from epidural neostigmine plus sufentanil bolus. In contrast, the TD did not significantly impair the ability to sit, stand up, or bend the knees. The ability to ambulate, however, was reduced (57% vs 82%; P = 0.04). In conclusion, a traditional lidocaine TD significantly enhances the analgesic effect from the epidural neostigmine plus sufentanil combination, but affects ambulation in early labor. PMID- 17122236 TI - Combined spinal epidural does not cause a higher sensory block than single shot spinal technique for cesarean delivery in laboring women. AB - BACKGROUND: The combined spinal epidural (CSE) technique has been shown to result in a higher sensory block than an equivalent single shot spinal (SSS) in women undergoing elective cesarean delivery. We tested whether this is true also in laboring women who may have variable epidural pressures. METHODS: We randomized 40 ASA I parturients in established labor for cesarean delivery into our double blind study. Group S (n = 20) intrathecally received 2 mL of 0.5% hyperbaric bupivacaine by SSS and group CS received CSE (n = 20) of an equivalent dose of hyperbaric bupivacaine. RESULTS: We found that similar maximal sensory blocks were achieved in both groups (group S: median T3 [min-max] T6-1 versus group CS: median T3 [min-max] T4-C7, P = 0.517). CONCLUSION: As compared with previous reports in nonlaboring parturients, the block characteristics of CSE in our study were indistinguishable from those of SSS in laboring parturients for cesarean delivery. PMID- 17122237 TI - The hemostatic profiles of patients with Type O and non-O blood after acute normovolemic hemodilution with 6% hydroxyethyl starch (130/0.4). AB - BACKGROUND: Individuals with Type O blood have been reported to have a tendency toward reduced Factor VIII and von Willebrand Factor (vWF) levels. If this is true, patients with Type O blood might be vulnerable to coagulopathy during acute normovolemic hemodilution using hydroxyethyl starch (HES), both from hemodilution as well as HES-related coagulopathy. METHODS: Thirty non-O and 15 type O ASA 1 or 2 patients scheduled for spinal surgery involving more than two spinal levels were enrolled for the study. After anesthesia induction, 30% of the estimated blood volume was removed, and the volume was simultaneously replaced with 6% HES (130/0.4). Coagulation profiles were measured before (T0) and 30 min after acute normovolemic hemodilution (T30). RESULTS: Factor VIII activity, vWF antigen levels (vWF:ag), and vWF ristocetin cofactor activity (vWF:RCof) were lower in the O group than in the non-O group before and after acute normovolemic hemodilution, and decreased below the normal range in the O group after acute normovolemic hemodilution. The decrease was beyond that expected from hemodilution alone. Maximum amplitude and coagulation index of the thromboelastogram decreased below the normal range in the O group after acute normovolemic hemodilution. The decrease in vWF:ag was related to the degree of blood loss, and was greater in patients in the O group. CONCLUSIONS: Patients with Type O blood may have increased coagulation compromise, and greater dilution of Factor VIII activity, vWF:ag, and vWF:RCof after acute normovolemic hemodilution with HES. PMID- 17122238 TI - Intraoperative infusion of amino acids induces anabolism independent of the type of anesthesia. AB - BACKGROUND: The infusion of dextrose in patients receiving epidural and light general anesthesia or general anesthesia alone failed to achieve a positive protein balance. We sought to verify the hypothesis that nutritional supplementation with i.v. amino acids induced a greater protein balance in patients receiving epidural blockade compared with those receiving general anesthesia. METHODS: Sixteen patients were randomly assigned to receive either general anesthesia with desflurane (control group) or general anesthesia combined with epidural analgesia (EDA group). A primed constant infusion of stable isotope tracers L-[1-(13)C]leucine and [6,6-(2)H2]glucose was started after a 32-h fast before surgery, (3 h of fasted state), and continued for 3 h during surgery during which amino acids were infused i.v. (fed state). RESULTS: Compared with the fasted state, the endogenous rate of appearance of leucine decreased to a similar extent in both groups, and protein synthesis increased, with no difference between the two groups. Leucine oxidation did not change in either group. After amino acids infusion, endogenous glucose production remained unchanged and glucose clearance decreased in both groups. Blood glucose, plasma cortisol, serum insulin, and glucagon concentrations increased to the same extent in both groups. CONCLUSIONS: Epidural anesthesia provided no additional benefit beyond the anabolism obtained with amino acids. PMID- 17122239 TI - The neural cell adhesion molecule antibody blocks cold water swim stress-induced analgesia and cell adhesion between lymphocytes and cultured dorsal root ganglion neurons. AB - BACKGROUND: Opioid-containing immune cells migrate in a site-directed manner into inflamed tissue and adhere to sensory nerve fibers. These cells release opioid peptides in close proximity to these fibers, thereby avoiding localized degradation by peptidases, and delivering opioid peptides proximal to opioid receptors to provide antinociception. METHODS: The effects of the anti-neural cell-adhesion molecule (anti-NCAM) were assessed on cold water swim stress induced antinociception in Wistar rats with Freund's adjuvant-induced inflammation of one hindpaw. Algesiometry was assessed for both thermal and mechanical stimuli. Cell adhesion experiments examining the effects of beta endorphin and antibodies to NCAM and intercellular cell adhesion molecule-1 and were performed on cultured dorsal root ganglion neurons and isolated lymphocytes. Lymphocyte binding was determined by fluorescence using calcein AM loaded into freshly isolated lymphocytes. RESULTS: The direct adhesion between lymphocytes and cultured sensory neurons was inhibited by anti-NCAM. This adhesion was also demonstrated to be opioid dependent, with lymphocyte adhesion to cultured sensory neurons reduced in the presence of 1 microM beta-endorphin, which was reversed by 100 microM naloxone. Moreover, anti-NCAM blocked cold-water-swim-induced analgesia in inflamed paws both to thermal and mechanical stimuli. However, anti NCAM did not affect fentanyl-induced antinociception. CONCLUSIONS: This study provides insight into the role of cell adhesion molecules in lymphocyte adhesion to sensory neurons and a link to immune-derived antinociception. PMID- 17122240 TI - Continuous femoral nerve analgesia after unilateral total knee arthroplasty: stimulating versus nonstimulating catheters. AB - BACKGROUND: Continuous femoral analgesia provides extended pain relief and improved functional recovery for total knee arthroplasty (TKA). Stimulating catheters may allow more accurate placement of catheters. METHODS: We performed a randomized prospective study to investigate the use of stimulating catheters versus nonstimulating catheters in 41 patients undergoing TKA. All patients received i.v. patient-controlled anesthesia for supplementary pain relief. The principal aim of the trial was to examine whether a stimulating catheter allowed the use of lesser amounts of local anesthetics than a nonstimulating catheter. The additional variables we examined included postoperative pain scores, opioid use, side effects, and acute functional orthopedic outcomes. RESULTS: Analgesia was satisfactory in both groups, but there were no statistically significant differences in the amount of ropivacaine administered; the median amount of ropivacaine given to patients in the stimulating catheter group was 8.2 mL/h vs 8.8 mL/h for patients with nonstimulating catheters, P = 0.26 (median difference 0.6; 95% confidence interval, -2.3 to 0.6). No significant differences between the treatment groups were noted for the amount of fentanyl dispensed by the i.v. patient-controlled anesthesia, numeric pain rating scale scores, acute functional orthopedic outcomes, side effects, or amounts of oral opioids consumed. CONCLUSION: The use of stimulating catheters in continuous femoral nerve blocks for TKA does not offer significant benefits over traditional nonstimulating catheters. PMID- 17122241 TI - The posterior approach to the sciatic nerve in the popliteal fossa: a comparison of single- versus double-injection technique. AB - We compared single-injection and double-injection of the sciatic nerve with nerve stimulation in the posterior popliteal approach using mepivacaine 1% in a prospective, randomized and single-blind study to evaluate effectiveness, delay of onset, and complications in patients undergoing foot and ankle surgery. In the single-injection group (Group S, n = 30), 25 mL of mepivacaine 1% was administered after eliciting foot inversion or plantar flexion. In the double injection group (Group D, n = 30), 12.5 mL of the solution was injected after eversion or dorsiflexion and 12.5 mL after plantar flexion of the foot. Mean differences (SD) between the two groups from onset time to complete sensory block were not significant (21.9 [14.2] min vs 22.1 [13.8] min) except for the superficial peroneal nerve block (18 [13] min vs 11.4 [7.5] min, Group S and D, respectively; P < 0.05) and, in Group D, between the superficial peroneal and tibial nerve blocks (11.4 [7.5] min vs 22.3 [11.3] min, respectively; P < 0.05). Complete analgesia was achieved in 77% of Group S patients and in 87% of Group D (P = 0.22). Complete analgesia of the deep peroneal nerve was achieved in 80% and 97% in Group S and D, respectively; P < 0.05. There were more paresthesias during block procedure in Group D (17% vs 40%) (P < 0.05). We conclude that double-nerve stimulation of the sciatic nerve gives similar complete onset times and overall success rate to single-nerve stimulation and more paresthesias during block performance. PMID- 17122242 TI - Use of magnetic resonance imaging to define the anatomical location closest to all three cords of the infraclavicular brachial plexus. AB - Infraclavicular techniques are often used to perform brachial plexus blocks. In our volunteer study we used magnetic resonance imaging to identify the brachial plexus and axillary vessels in a sagittal plane corresponding to the lateral sagittal infraclavicular block. In 20 volunteers, all cords were positioned within 2 cm from the artery approximately within 2/3 of a circle. We derived an injection site that was closest to all cords, cranio-posterior and adjacent to the axillary artery. We conclude that this knowledge may be useful for the performance of infraclavicular blocks aided by ultrasound. However, our proposals should be tested by clinical studies. PMID- 17122246 TI - Predictive performance of three multivariate difficult tracheal intubation models: a double-blind, case-control study. PMID- 17122247 TI - Rapid-sequence induction: rocuronium or suxamethonium? PMID- 17122248 TI - The ideal dose of succinylcholine for tracheal intubation. PMID- 17122249 TI - Closed-circuit anesthesia is neither dead nor forgotten. PMID- 17122250 TI - In defense of the continued use of carbon dioxide absorbents. PMID- 17122251 TI - Is ephedrine the best vasopressor for treating spinal anesthesia-induced hypotension in patients with pre-eclampsia? PMID- 17122252 TI - A contraindication to using local anesthetic solution for expanding the epidural space. PMID- 17122253 TI - Prolonged activated partial thromboplastin time in patients with antiphospholipid antibody syndrome. PMID- 17122254 TI - Chemical dependency treatment outcomes of residents. PMID- 17122255 TI - A testable hypothesis that may explain the morbidity and mortality caused by surgical stress. PMID- 17122256 TI - What is the definite merit of clonidine premedication in patients with sleep apnea? PMID- 17122257 TI - The use of epidural analgesia in cardiac surgery should be encouraged. PMID- 17122258 TI - Tourniquet injuries, implied causality, babies, and bathwater. PMID- 17122259 TI - Unusual defect in a double-lumen endotracheal tube. PMID- 17122260 TI - Run out of O2? Use transport O2. PMID- 17122261 TI - Anesthetic management during bronchoscopic removal of a unique, friable foreign body. PMID- 17122262 TI - Tramadol, vecuronium, and thoracic epidural ropivacaine combined with sevoflurane anesthesia in a patient with human T-lymphotropic virus type 1-associated myelopathy. PMID- 17122263 TI - Demographics of nonagenarians and centenarians with a hip fracture. PMID- 17122264 TI - Delayed subarachnoid hematoma after continuous epidural anesthesia in a patient without risk factors: good outcome after late laminectomy. PMID- 17122265 TI - The partial CO2 rebreathing cardiac output measurement during laparoscopic resection of bilateral pheochromocytoma. PMID- 17122266 TI - Intraoperative kinking of polyvinyl endotracheal tubes. PMID- 17122267 TI - Coronary artery spasm as a cause of droperidol-induced ventricular fibrillation. PMID- 17122268 TI - Ventilatory support during neuraxial blockade using bilevel positive airway pressure in a patient with severe respiratory compromise. PMID- 17122269 TI - More cockroaches. PMID- 17122270 TI - Secure the torpedoes! PMID- 17122271 TI - Warming crystalloid fluid for intravenous infusion: how effective is a fluid warming cabinet? PMID- 17122272 TI - The effects of electromyographic activity on the bispectral index during combined anesthesia. PMID- 17122273 TI - Does atropine reduce the risk of propofol-induced cardiovascular depression? PMID- 17122274 TI - Regional cerebral oxyhemoglobin saturation-guided resuscitation. PMID- 17122275 TI - Fluid restriction and major surgery. PMID- 17122276 TI - Elevated burst suppression ratio: the possible role of hypoxemia. PMID- 17122277 TI - An unusual cause of a "missing tip" in an Arrow Flextip Plus epidural catheter. PMID- 17122278 TI - Spinal subdural hematoma: a pre-eclamptic patient with a spinal arteriovenous malformation. PMID- 17122279 TI - Anesthetic issues with a hyperextended cervical spine in a child with Emery Dreifuss syndrome. PMID- 17122280 TI - Transient but profound reduction of bispectral index values after tourniquet deflation: did the BIS detect an alteration of brain electrocortical activity? PMID- 17122281 TI - Aprotinin and sevoflurane do not affect renal function during single-ventricle palliative surgery. PMID- 17122282 TI - A survey of ophthalmic anesthetists on managing pacemakers and implanted cardiac defibrillators. PMID- 17122283 TI - Treatment of postherpetic neuralgia with intravenous administration of vitamin C. PMID- 17122284 TI - Drop foot after pediatric urological surgery under general and epidural anesthesia. PMID- 17122285 TI - Incorrect shunt placement due to anatomic variations of the aortic arch during carotid endarterectomy: a rare cause of perioperative ischemia? PMID- 17122286 TI - Using dexmedetomidine to manage patients with cocaine and opioid withdrawal, who are undergoing cerebral angioplasty for cerebral vasospasm. PMID- 17122287 TI - Delayed recovery after short-duration, general anesthesia in a patient chronically treated with clozapine. PMID- 17122288 TI - Recovering a detached Univent endobronchial balloon. PMID- 17122289 TI - Fixation of an oral tracheal tube to the maxilla in maxillofacial surgery. PMID- 17122290 TI - Stuck tracheostomy button: an unusual complication. PMID- 17122291 TI - Increased propensity to bruising in red-haired females: a possible role for von Willebrand factor? PMID- 17122292 TI - Tracheal intubation using a classic laryngeal mask airway, Frova introducer, and pediatric bronchoscope. PMID- 17122293 TI - Pediatric computed tomographic scan with anesthesia: what the anesthesiologist should know. PMID- 17122294 TI - The dissection of a reinforced endotracheal tube causing near-fatal intraoperative airway obstruction. PMID- 17122295 TI - Combined spinal-epidural anesthesia for a child with Freeman-Sheldon syndrome with difficult airway. PMID- 17122296 TI - Acute cerebellar stroke after inadvertent cannulation and pulmonary artery catheter placement in the right vertebral artery. PMID- 17122297 TI - Desflurane vaporizer: two hazardous incidents. PMID- 17122298 TI - Near-fatal ventricular arrhythmias because of central venous pressure catheter tip migration during laparoscopic hemicolectomy. PMID- 17122299 TI - Respiratory depression caused by remifentanil infusion for postoperative pain control. PMID- 17122300 TI - Retrograde tracheal intubation in a patient with a halo traction device. PMID- 17122301 TI - Tube exchanger for laryngeal mask-based percutaneous tracheostomy in the intensive care unit. PMID- 17122302 TI - Combined general and regional anesthetic in a child with aromatic L-amino acid decarboxylase deficiency. PMID- 17122303 TI - Radiation exposure in children. PMID- 17122304 TI - Glycine encephalopathy and delayed emergence from anesthesia. PMID- 17122305 TI - Misdiagnosis of perioperative myocardial ischemia: the effects of electrocardiogram filtering. PMID- 17122306 TI - Misdirected pulmonary artery catheter. PMID- 17122307 TI - Neonatal airway management in occipital encephalocele. PMID- 17122308 TI - An approach to infraclavicular brachial plexus block: the highest point of the shoulder as a reference. PMID- 17122309 TI - A soft cervical collar may help prevent neck sprain or nerve injury from occurring after surgery in the lateral decubitus position. PMID- 17122310 TI - Recovery of electromyographic activity after transection and surgical repair of the rat sciatic nerve. AB - The recovery of soleus (SOL), gastrocnemius (GAS), and tibialis anterior (TA) electromyographic activity (EMG) after transection and surgical repair of the sciatic nerve was studied in Sprague-Dawley rats using chronically implanted stimulation and recording electrodes. Spontaneous EMG activity in SOL and GAS and direct muscle (M) responses to posterior tibial nerve stimulation persisted for < or =2 days after sciatic nerve transection, but SOL and GAS H-reflexes disappeared immediately. Spontaneous EMG activity began to return 2-3 wk after transection, rose nearly to pretransection levels by 60 days, and persisted for the duration of the study period (120 days). Recovery of stimulus-evoked EMG responses began about 30 days after sciatic nerve transection as multiple small responses with a wide range of latencies. Over time, the latencies of these fractionated responses shortened, their amplitudes increased, and they merged into a distinct short-latency component (the putative M response) and a distinct long-latency component (the putative H-reflex). The extent of recovery of stimulation-evoked EMG was modest: even 100 days after sciatic nerve transection, the responses were still much smaller than those before transection. Similar gradual development of responses to posterior tibial nerve stimulation was also seen in TA, suggesting that some regenerating fibers sent branches into both tibial and common peroneal nerves. PMID- 17122311 TI - Contrast gain control is drift-rate dependent: an informational analysis. AB - Neurons in the visual cortex code relative changes in illumination (contrast) and adapt their sensitivities to the visual scene by centering the steepest regions of their sigmoidal contrast response functions (CRFs: spike rate as a function of contrast) on the prevailing contrast. The influence of this contrast gain control has not been reported at nonoptimal drift rates. We calculated the Fisher information contained in the CRFs of halothane-anesthetized cats. Fisher information gives a measure of the accuracy of contrast representations based on the ratio of the square of the steepness of the CRF and the spike-rate dependency of the spiking variance. Variance increases with spike rate, so Fisher information is maximal where the CRF is steep and spike rates are low. Here, we show that the contrast at which the maximal Fisher information (C(MFI)) occurs for each adapting drift rate is at a fixed level above the adapting contrast. For adapting contrasts of 0 to 0.32 the relationship between C(MFI) and adapting contrast is well described by a straight line with a slope close to 1. The intercept of this line on the C(MFI)-axis is drift-rate dependent, although the slope is not. At high drift rates relative to each cell's peak the C(MFI) offset is higher than that for low drift rates. The results show that the contrast coding strategy in visual cortex maximizes accuracy for contrasts above the prevailing contrast in the environment for all drift rates. We argue that tuning the system for accuracy at contrasts above the prevailing value is optimal for viewing natural scenes. PMID- 17122312 TI - Kappa opioid inhibition of somatodendritic dopamine inhibitory postsynaptic currents. AB - In the midbrain, dopamine neurons can release dopamine somatodendritically. This results in an inhibitory postsynaptic current (IPSC) within adjacent dopamine cells that occurs by the activation of inhibitory D(2) autoreceptors. Kappa, but not mu/delta, opioid receptors inhibit this IPSC. The aim of the present study was to determine the mechanism by which kappa-opioid receptors inhibit the dopamine IPSC. In both the ventral tegmental area (VTA) and substantia nigra compacta (SNc) the kappa-receptor agonist U69593 inhibited the IPSC, but not the current induced by the exogenous iontophoretic application of dopamine. The endogenous peptide dynorphin A (1-13) also inhibited IPSCs in the VTA and SNc, but also the dopamine iontophoretic current in the VTA. Although both kappa agonists induced a postsynaptic outward current in the VTA, the current induced by dynorphin was dramatically larger. This suggests that the decrease in iontophoretic dopamine current was the result of occlusion. Occlusion alone, however, could not completely account for suppression of the IPSC. The kappa opioid inhibition of the IPSC was not affected by global increases or decreases in dopamine cell activity within the slice. These findings suggest that, although kappa opioid receptors can hyperpolarize dopamine neurons, they also suppress dopamine release by direct actions at the release site. The results thus demonstrate both pre- and postsynaptic actions of kappa receptor agonists. The actions of dynorphin indicate that VTA dopamine cells are selectively regulated by kappa receptors. PMID- 17122313 TI - Response of vestibular-nerve afferents to active and passive rotations under normal conditions and after unilateral labyrinthectomy. AB - We investigated the possible contribution of signals carried by vestibular-nerve afferents to long-term processes of vestibular compensation after unilateral labyrinthectomy. Semicircular canal afferents were recorded from the contralesional nerve in three macaque monkeys before [horizontal (HC) = 67, anterior (AC) = 66, posterior (PC) = 50] and 1-12 mo after (HC = 192, AC = 86, PC = 57) lesion. Vestibular responses were evaluated using passive sinusoidal rotations with frequencies of 0.5-15 Hz (20-80 degrees /s) and fast whole-body rotations reaching velocities of 500 degrees /s. Sensitivities to nonvestibular inputs were tested by: 1) comparing responses during active and passive head movements, 2) rotating the body with the head held stationary to activate neck proprioceptors, and 3) encouraging head-restrained animals to attempt to make head movements that resulted in the production of neck torques of < or =2 Nm. Mean resting discharge rate before and after the lesion did not differ for the regular, D (dimorphic)-irregular, or C (calyx)-irregular afferents. In response to passive rotations, afferents showed no change in sensitivity and phase, inhibitory cutoff, and excitatory saturation after unilateral labyrinthectomy. Moreover, head sensitivities were similar during voluntary and passive head rotations and responses were not altered by neck proprioceptive or efference copy signals before or after the lesion. The only significant change was an increase in the proportion of C-irregular units postlesion, accompanied by a decrease in the proportion of regular afferents. Taken together, our findings show that changes in response properties of the vestibular afferent population are not likely to play a major role in the long-term changes associated with compensation after unilateral labyrinthectomy. PMID- 17122314 TI - Electrophysiological differences between neurogliaform cells from monkey and rat prefrontal cortex. AB - Current dogma holds that a canonical cortical circuit is formed by cellular elements that are basically identical across species. However, detailed and direct comparisons between species of specific elements of this circuit are limited in number. In this study, we compared the morphological and physiological properties of neurogliaform (NGF) inhibitory neurons in the prefrontal cortex (PFC) of macaque monkeys and rats. In both species, NGF cells were readily identified based on their distinctive morphological features. Indeed, monkey NGF cells had only a few morphological features that differed from rat, including a larger soma, a greater number of dendrites, and a more compact axonal field. In contrast, whole cell recordings of the responses to injected current steps revealed important differences between monkey and rat NGF cells. Monkey NGF cells consistently generated a short-latency first spike riding on an initial depolarizing hump, whereas in rat NGF cells, the first spike appeared after a substantial delay riding on a depolarizing ramp not seen in monkey NGF cells. Thus although rat NGF cells are traditionally classified as late-spiking cells, monkey NGF cells did not meet this physiological criterion. In addition, NGF cells in monkey appeared to be more excitable than those in rat because they displayed a higher input resistance, a lower spike threshold, and a higher firing frequency. Finally, NGF cells in monkey showed a more prominent spike-frequency adaptation as compared with rat. Our findings indicate that the canonical cortical circuit differs in at least some aspects of its constituent elements across species. PMID- 17122315 TI - Agonist-dependent postsynaptic effects of opioids on miniature excitatory postsynaptic currents in cultured hippocampal neurons. AB - Although chronic treatment with morphine is known to alter the function and morphology of excitatory synapses, the effects of other opioids on these synapses are not clear. Here we report distinct effects of several opioids (morphine, [d ala(2),me-phe(4),gly(5)-ol]enkephalin (DAMGO), and etorphine) on miniature excitatory postsynaptic currents (mEPSCs) in cultured hippocampal neurons: 1) chronic treatment with morphine for >3 days decreased the amplitude, frequency, rise time and decay time of mEPSCs. In contrast, "internalizing" opioids such as etorphine and DAMGO increased the frequency of mEPSCs and had no significant effect on the amplitude and kinetics of mEPSCs. These results demonstrate that different opioids can have distinct effects on the function of excitatory synapses. 2) mu opioid receptor fused with green fluorescence protein (MOR-GFP) is clustered in dendritic spines in most hippocampal neurons but is concentrated in axon-like processes in striatal and corticostriatal nonspiny neurons. It suggests that MORs might mediate pre- or postsynaptic effects depending on cell types. 3) Neurons were cultured from MOR knock-out mice and were exogenously transfected with MOR-GFP. Chronic treatment with morphine suppressed mEPSCs only in neurons that contained postsynaptic MOR-GFP, indicating that opioids can modulate excitatory synaptic transmission postsynaptically. 4) Morphine acutely decreased mEPSC amplitude in neurons expressing exogenous MOR-GFP but had no effect on neurons expressing GFP. It indicates that the low level of endogenous MORs could only allow slow opioid-induced plasticity of excitatory synapses under normal conditions. 5) A theoretical model suggests that morphine might affect the function of spines by decreasing the electrotonic distance from synaptic inputs to the soma. PMID- 17122316 TI - Chlorpromazine alters cochlear mechanics and amplification: in vivo evidence for a role of stiffness modulation in the organ of corti. AB - Although prestin-mediated outer hair cell (OHC) electromotility provides mechanical force for sound amplification in the mammalian cochlea, proper OHC stiffness is required to maintain normal electromotility and to transmit mechanical force to the basilar membrane (BM). To investigate the in vivo role of OHC stiffness in cochlear amplification, chlorpromazine (CPZ), an antipsychotic drug that alters OHC lateral wall biophysics, was infused into the cochleae in living guinea pigs. The effects of CPZ on cochlear amplification and OHC electromotility were observed by measuring the acoustically and electrically evoked BM motions. CPZ significantly reduced cochlear amplification as measured by a decline of the acoustically evoked BM motion near the best frequency (BF) accompanied by a loss of nonlinearity and broadened tuning. It also substantially reduced electrically evoked BM vibration near the BF and at frequencies above BF (< or =80 kHz). The high-frequency notch (near 50 kHz) in the electrically evoked BM response shifted toward higher frequency in a CPZ concentration-dependent manner with a corresponding phase change. In contrast, salicylate resulted in a shift in this notch toward lower frequency. These results indicate that CPZ reduces OHC-mediated cochlear amplification probably via its effects on the mechanics of the OHC plasma membrane rather than via a direct effect on the OHC motor, prestin. Through modeling, we propose that with a combined OHC somatic and hair bundle forcing, the upward-shift of the approximately 50-kHz notch in the electrically-evoked BM motion may indicate stiffness increase of the OHCs that is responsible for the reduced cochlear amplification. PMID- 17122317 TI - Reward value coding distinct from risk attitude-related uncertainty coding in human reward systems. AB - When deciding between different options, individuals are guided by the expected (mean) value of the different outcomes and by the associated degrees of uncertainty. We used functional magnetic resonance imaging to identify brain activations coding the key decision parameters of expected value (magnitude and probability) separately from uncertainty (statistical variance) of monetary rewards. Participants discriminated behaviorally between stimuli associated with different expected values and uncertainty. Stimuli associated with higher expected values elicited monotonically increasing activations in distinct regions of the striatum, irrespective of different combinations of magnitude and probability. Stimuli associated with higher uncertainty (variance) elicited increasing activations in the lateral orbitofrontal cortex. Uncertainty-related activations covaried with individual risk aversion in lateral orbitofrontal regions and risk-seeking in more medial areas. Furthermore, activations in expected value-coding regions in prefrontal cortex covaried differentially with uncertainty depending on risk attitudes of individual participants, suggesting that separate prefrontal regions are involved in risk aversion and seeking. These data demonstrate the distinct coding in key reward structures of the two basic and crucial decision parameters, expected value, and uncertainty. PMID- 17122318 TI - Sensorimotor integration in S2, PV, and parietal rostroventral areas of the human sylvian fissure. AB - We explored cortical fields on the upper bank of the Sylvian fissure using functional magnetic resonance imaging (fMRI) and magnetoencephalography (MEG) to measure responses to two stimulus conditions: a tactile stimulus applied to the right hand and a tactile stimulus with an additional movement component. fMRI data revealed bilateral activation in S2/PV in response to tactile stimulation alone and source localization of MEG data identified a peak latency of 122 ms in a similar location. During the tactile and movement condition, fMRI revealed bilateral activation of S2/PV and an anterior field, while MEG data contained one source at a location identical to the tactile-only condition with a latency of 96 ms and a second rostral source with a longer latency (136 ms). Furthermore, Region-of-interest analysis of fMRI data identified increased bilateral activation in S2/PV and the rostral area in the tactile and movement condition compared with the tactile only condition. An area of cortex immediately rostral to S2/PV in monkeys has been called the parietal rostroventral area (PR). Based on location, latency, and conditions under which this field was active, we have termed the rostral area of human cortex PR as well. These findings indicate that humans, like non-human primates, have a cortical field rostral to PV that processes proprioceptive inputs, both S2/PV and PR play a role in somatomotor integration necessary for manual exploration and object discrimination, and there is a temporal hierarchy of processing with S2/PV active prior to PR. PMID- 17122319 TI - Multiple sensory cues underlying the perception of translation and path. AB - The translational linear vestibuloocular reflex compensates most accurately for high frequencies of head translation, with response magnitude decreasing with declining stimulus frequency. However, studies of the perception of translation typically report robust responses even at low frequencies or during prolonged motion. This inconsistency may reflect the incorporation of nondirectional sensory information associated with the vibration and noise that typically accompany translation, into motion perception. We investigated the perception of passive translation in humans while dissociating nondirectional cues from actual head motion. In a cue-dissociation experiment, interaural (IA) motion was generated using either a linear sled, the mechanics of which generated noise and vibration cues that were correlated with the motion profile, or a multiaxis technique that dissociated these cues from actual motion. In a trajectory-shift experiment, IA motion was interrupted by a sudden change in direction (+/-30 degrees diagonal) that produced a change in linear acceleration while maintaining sled speed and therefore mechanical (nondirectional) cues. During multi-axis cue dissociation trials, subjects reported erroneous translation perceptions that strongly reflected the pattern of nondirectional cues, as opposed to nearly veridical percepts when motion and nondirectional cues coincided. During trajectory-shift trials, subjects' percepts were initially accurate, but erroneous following the direction change. Results suggest that nondirectional cues strongly influence the perception of linear motion, while the utility of cues directly related to translational acceleration is limited. One key implication is that "path integration" likely involves complex mechanisms that depend on nondirectional and contextual self-motion cues in support of limited and transient otolith-dependent acceleration input. PMID- 17122320 TI - Spastic tail muscles recover from myofiber atrophy and myosin heavy chain transformations in chronic spinal rats. AB - Without intervention after spinal cord injury (SCI), paralyzed skeletal muscles undergo myofiber atrophy and slow-to-fast myofiber type transformations. We hypothesized that chronic spasticity-associated neuromuscular activity after SCI would promote recovery from such deleterious changes. We examined segmental tail muscles of chronic spinal rats with long-standing tail spasticity (7 mo after sacral spinal cord transection; older chronic spinals), chronic spinal rats that experienced less spasticity early after injury (young chronic spinals), and rats without spasticity after transection and bilateral deafferentation (spinal isolated). These were compared with tail muscles of age-matched normal rats. Using immunohistochemistry, we observed myofiber distributions of 15.9 +/- 3.5% type I, 18.7 +/- 10.7% type IIA, 60.8 +/- 12.6% type IID(X), and 2.3 +/- 1.3% type IIB (means +/- SD) in young normals, which were not different in older normals. Young chronic spinals demonstrated transformations toward faster myofiber types with decreased type I and increased type IID(X) paralleled by atrophy of all myofiber types compared with young normals. Spinal isolated rats also demonstrated decreased type I myofiber proportions and increased type II myofiber proportions, and severe myofiber atrophy. After 4 mo of complete spasticity (older chronic spinals), myofiber type transformations were reversed, with no significant differences in type I, IIA, IID(X), or IIB proportions compared with age-matched normals. Moreover, after this prolonged spasticity, type I, IIA, and IIB myofibers recovered from atrophy, and type IID(X) myofibers partially recovered. Our results indicate that early after transection or after long-term spinal isolation, relatively inactive tail myofibers atrophy and transform toward faster myofiber types. However, long-term spasticity apparently produces neuromuscular activity that promotes recovery of myofiber types and myofiber sizes. PMID- 17122321 TI - Distinct roles for glycine and GABA in shaping the response properties of neurons in the superior paraolivary nucleus of the rat. AB - The superior paraolivary nucleus (SPON) is a prominent periolivary cell group of the superior olivary complex. SPON neurons use gamma-aminobutyric acid (GABA) as their neurotransmitter and are contacted by large numbers of glycinergic and GABAergic punctate profiles, representing a dense inhibitory innervation from the medial nucleus of the trapezoid body (MNTB) and from collaterals of SPON axons, respectively. SPON neurons have low rates of spontaneous activity, respond preferentially to the offset of pure tones, and phase-lock to amplitude-modulated tones. To determine the roles of glycine and GABA in shaping SPON responses, we recorded from single units in the SPON of anesthetized rats before, during, and after application of the glycine receptor antagonist strychnine, the GABA(A) receptor antagonist bicuculline, or both drugs applied simultaneously. Strychnine caused a major increase in spike counts during the stimulus presentation, followed by the disappearance of offset spikes. In half of the recorded units, bicuculline caused moderately increased firing during the stimulus. However, in 86% of units bicuculline also caused a large increase in the magnitude of the offset response. Application of the drug cocktail caused increased spontaneous activity, dramatically increased spike counts during the stimulus presentation, and eliminated the offset response in most units. We conclude that glycinergic inhibition from the MNTB suppresses SPON spiking during sound stimulation and is essential in generating offset responses. GABAergic inhibition, presumably from intrinsic SPON collaterals, plays a subtler role, contributing in some cells to suppression of firing during the stimulus and in most cells to restrict firing after stimulus offset. PMID- 17122322 TI - "Motor oblique effect": perceptual direction discrimination and pointing to memorized visual targets share the same preference for cardinal orientations. AB - In previous studies we observed a pattern of systematic directional errors when humans pointed to memorized visual target locations in two-dimensional (2-D) space. This directional error was also observed in the initial direction of slow movements toward visual targets or movements to kinesthetically defined targets in 2-D space. In this study we used a perceptual experiment where subjects decide whether an arrow points in the direction of a visual target in 2-D space and observed a systematic distortion in direction discrimination known as the "oblique effect." More specifically, direction discrimination was better for cardinal directions than for oblique. We then used an equivalent measure of direction discrimination in a task where subjects pointed to memorized visual target locations and showed the presence of a motor oblique effect. We finally modeled the oblique effect in the perceptual and motor task using a quadratic function. The model successfully predicted the observed direction discrimination differences in both tasks and, furthermore, the parameter of the model that was related to the shape of the function was not different between the motor and the perceptual tasks. We conclude that a similarly distorted representation of target direction is present for memorized pointing movements and perceptual direction discrimination. PMID- 17122323 TI - Heterogeneity of phasic cholinergic signaling in neocortical neurons. AB - Acetylcholine (ACh) is a neurotransmitter critical for normal cognition. Here we demonstrate heterogeneity of cholinergic signaling in neocortical neurons in the rat prefrontal, somatosensory, and visual cortex. Focal ACh application (100 muM) inhibited layer 5 pyramidal neurons in all cortical areas via activation of an apamin-sensitive SK-type calcium-activated potassium conductance. Cholinergic inhibition was most robust in prefrontal layer 5 neurons, where it relies on the same signal transduction mechanism (M1-like receptors, IP(3)-dependent calcium release, and SK-channels) as exists in somatosensory pyramidal neurons. Pyramidal neurons in layer 2/3 were less responsive to ACh, but substantial apamin sensitive inhibitory responses occurred in deep layer 3 neurons of the visual cortex. ACh was only inhibitory when presented near the somata of layer 5 pyramidal neurons, where repetitive ACh applications generated discrete inhibitory events at frequencies of up to approximately 0.5 Hz. Fast-spiking (FS) nonpyramidal neurons in all cortical areas were unresponsive to ACh. When applied to non-FS interneurons in layers 2/3 and 5, ACh generated mecamylamine-sensitive nicotinic responses (38% of cells), apamin-insensitive hyperpolarizing responses, with or without initial nicotinic depolarization (7% of neurons), or no response at all (55% of cells). Responses in interneurons were similar across cortical layers and regions but were correlated with cellular physiology and the expression of biochemical markers associated with different classes of nonpyramidal neurons. Finally, ACh generated nicotinic responses in all layer 1 neurons tested. These data demonstrate that phasic cholinergic input can directly inhibit projection neurons throughout the cortex while sculpting intracortical processing, especially in superficial layers. PMID- 17122325 TI - Modified sensory processing in the barrel cortex of the adult mouse after chronic whisker stimulation. AB - Chronic stimulation of a mystacial whisker follicle for 24 h induces structural and functional changes in layer IV of the corresponding barrel, with an insertion of new inhibitory synapses on spines and a depression of neuronal responses to the stimulated whisker. Under urethane anesthesia, we analyzed how sensory responses of single units are affected in layer IV and layers II & III of the stimulated barrel column as well as in adjacent columns. In the stimulated column, spatiotemporal characteristics of the activation evoked by the stimulated whisker are not altered, although spontaneous activity and response magnitude to the stimulated whisker are decreased. The sensitivity of neurons for the deflection of this whisker is not altered but the dynamic range of the response is reduced as tested by varying the amplitude and repetition rate of the deflection. Responses to deflection of nonstimulated whiskers remain unaltered with the exception of in-row whisker responses that are depressed in the column corresponding to the stimulated whisker. In adjacent nonstimulated columns, neuronal activity remains unaltered except for a diminished response of units in layer II/III to deflection of the stimulated whisker. From these results we propose that an increased inhibition within the stimulated barrel reduced the magnitude of its excitatory output and accordingly the flow of excitation toward layers II & III and the subsequent spread into adjacent columns. In addition, the period of uncorrelated activity between pathways from the stimulated and nonstimulated whiskers weakens synaptic inputs from in-row whiskers in the stimulated barrel column. PMID- 17122324 TI - Dual diffusion model for single-cell recording data from the superior colliculus in a brightness-discrimination task. AB - Monkeys made saccades to one of two peripheral targets based on the brightness of a central stimulus. Task difficulty was manipulated by varying the ratio of stimulus black-and-white pixels. Correct response probability for two monkeys varied directly with difficulty. Deep layer SC neurons exhibited robust presaccadic activity the magnitude of which was unaffected by task difficulty when the stimulus specified a saccade toward a target within the neuron's response field. Activity after stimuli specifying saccades to targets outside the response field was affected by task difficulty, increasing as the task became more difficult. A quantitative model derived from studies of human decision making was fit to the behavioral data. The model assumes that information from the stimulus drives two independent diffusion processes. Simulated paths from the model were compared with neuron activity, assuming that firing rate is linearly related to position in the accumulation process. The firing rate data show delayed availability of discriminative information for fast, intermediate, and slow decisions when activity is aligned on the stimulus and very small differences in discriminative information when aligned on the saccade. The model produces exactly these patterns of results. The accumulation process is highly variable, allowing the process both to make errors, as is the case for the behavioral performance, and also to account for the firing rate results. Thus the dual diffusion model provides a quantitative account for both the behavior in a simple decision-making task as well as the patterns of activity in competing populations of neurons. PMID- 17122326 TI - Direct comparison of the task-dependent discharge of M1 in hand space and muscle space. AB - Since its introduction in the early 1980s, the concept of a "preferred direction" for neuronal discharge has proven to be a powerful means of studying motor areas of the brain. In the current paper, we introduce the concept of a "muscle-space" preferred direction (PD(M)) that is analogous to the familiar hand-space preferred direction (PD(H)). PD(M) reflects the similarity between the discharge of a given neuron and the activity of each muscle in much the way that PD(H) reflects the similarity of discharge with motion along each of the three Cartesian coordinate axes. We used PD(M) to analyze the data recorded from neurons in the primary motor cortex (M1) of three different monkeys. The monkeys performed center-out movements within two different cubical workspaces centered either to the left or right of the monkey's shoulder while we simultaneously recorded neuronal discharge, muscle activity, and limb orientation. We calculated preferred directions in both hand space and muscle space, and computed the angles between these vectors under a variety of conditions. PDs for different neurons were broadly distributed throughout both hand space and muscle space, but the muscle-space vectors appeared to form clusters of functionally similar neurons. In general, repeated estimates of PD(M) were more stable over time than were similar estimates of PD(H). Likewise, there was less change in PD(M) than in PD(H) for data recorded from the two different workspaces. However, although a majority of neurons had this muscle-like property, a significant minority was more stable in Cartesian hand space, reflecting a heterogeneity of function within M1. PMID- 17122327 TI - Effect of dietary hempseed intake on cardiac ischemia-reperfusion injury. AB - Polyunsaturated fatty acids (PUFAs) have significant, cardioprotective effects against ischemia. Hempseed contains a high proportion of the PUFAs linoleic acid (LA) and alpha-linolenic acid (ALA), which may have opposing effects on postischemic heart performance. There are no reported data concerning the cardiovascular effects of dietary hempseed intake. A group of 40 male Sprague Dawley rats were distributed evenly into four groups that were fed for 12 wk a normal rat chow supplemented with hempseed (5% and 10%), palm oil (1%), or a 10% partially delipidated hempseed that served as a control. Plasma ALA and gamma linolenic acid levels were significantly elevated in the rats that were fed a 5% or 10% hempseed-supplemented diet, but in heart tissue only ALA levels were significantly elevated in the rats fed these diets compared with control. After the dietary interventions were completed, postischemic heart performance was evaluated by measuring developed tension, resting tension, the rates of tension development and relaxation, and the number of extrasystoles. Hearts from rats fed a hempseed-supplemented diet exhibited significantly better postischemic recovery of maximal contractile function and enhanced rates of tension development and relaxation during reperfusion than hearts from the other groups. These hearts, however, were not protected from the occurrence of extrasystoles, nor were the increases in resting tension altered during ischemia or reperfusion as a function of any dietary intervention. Our data demonstrate that dietary hempseed can provide significant cardioprotective effects during postischemic reperfusion. This appears to be due to its highly enriched PUFA content. PMID- 17122328 TI - Lysophosphatidic acid induces endothelial cell death by modulating the redox environment. AB - Oxidant stress plays a significant role in hypoxic-ischemic injury to the susceptible microvascular endothelial cells. During oxidant stress, lysophosphatidic acid (LPA) concentrations increase. We explored whether LPA caused cytotoxicity to neuromicrovascular cells and the potential mechanisms thereof. LPA caused a dose-dependent death of porcine cerebral microvascular as well as human umbilical vein endothelial cells; cell death appeared oncotic rather than apoptotic. LPA-induced cell death was mediated via LPA(1) receptor, because the specific LPA(1) receptor antagonist THG1603 fully abrogated LPA's effects. LPA decreased intracellular GSH levels and induced a p38 MAPK/JNK dependent inducible nitric oxide synthase (NOS) expression. Pretreatment with the antioxidant GSH precursor N-acetyl-cysteine (NAC), as well as with inhibitors of NOS [N(omega)-nitro-l-arginine (l-NNA); 1400W], significantly prevented LPA induced endothelial cell death (in vitro) to comparable extents; as expected, p38 MAPK (SB203580) and JNK (SP-600125) inhibitors also diminished cell death. LPA did not increase indexes of oxidation (isoprostanes, hydroperoxides, and protein nitration) but did augment protein nitrosylation. Endothelial cytotoxicity by LPA in vitro was reproduced ex vivo in brain and in vivo in retina; THG1603, NAC, l NNA, and combined SB-203580 and SP600125 prevented the microvascular rarefaction. Data implicate novel properties for LPA as a modulator of the cell redox environment, which partakes in endothelial cell death and ensued neuromicrovascular rarefaction. PMID- 17122329 TI - Effect of arterial oxygenation on quadriceps fatigability during isolated muscle exercise. AB - The effect of various levels of oxygenation on quadriceps muscle fatigability during isolated muscle exercise was assessed in six male subjects. Twitch force (Q(tw)) was assessed using supramaximal magnetic femoral nerve stimulation. In experiment 1, maximal voluntary contraction (MVC) and Q(tw) of resting quadriceps muscle were measured in normoxia [inspired O(2) fraction (Fi(O(2))) = 0.21, percent arterial O(2) saturation (Sp(O(2))) = 98.4%, estimated arterial O(2) content (Ca(O(2))) = 20.8 ml/dl], acute hypoxia (Fi(O(2)) = 0.11, Sp(O(2)) = 74.6%, Ca(O(2)) = 15.7 ml/dl), and acute hyperoxia (Fi(O(2)) = 1.0, Sp(O(2)) = 100%, Ca(O(2)) = 22.6 ml/dl). No significant differences were found for MVC and Q(tw) among the three Fi(O(2)) levels. In experiment 2, the subjects performed three sets of nine, intermittent, isometric, unilateral, submaximal quadriceps contractions (62% MVC followed by 1 MVC in each set) while breathing each Fi(O(2)). Q(tw) was assessed before and after exercise, and myoelectrical activity of the vastus lateralis was obtained during exercise. The percent reduction of twitch force (potentiated Q(tw)) in hypoxia (-27.0%) was significantly (P < 0.05) greater than in normoxia (-21.4%) and hyperoxia ( 19.9%), as were the changes in intratwitch measures of contractile properties. The increase in integrated electromyogram over the course of the nine contractions in hypoxia (15.4%) was higher (P < 0.05) than in normoxia (7.2%) or hyperoxia (6.7%). These results demonstrate that quadriceps muscle fatigability during isolated muscle exercise is exacerbated in acute hypoxia, and these effects are independent of the relative exercise intensity. PMID- 17122330 TI - Deficiency in angiotensin AT1a receptors prevents diabetes-induced hypertension. AB - The renin-angiotensin system has been implicated in the etiology of the cardiovascular complications of diabetes. Our studies extend these findings to show a specific role for angiotensin AT1a receptors in mediating diabetes-induced hypertension. Male angiotensin AT1a knockout (AT1aKO) and wild-type (AT1aWT) mice with arterial telemetric catheters were injected with streptozotocin (STZ; 150 mg/kg ip). The STZ dose was selected on the basis of a dose-response experiment in C57/BL mice. Blood glucose, water intake, body weight, blood pressure (BP), and heart rate (HR) were measured over a 2-wk period. Estimates of BP and HR variance (BPV and HRV) and their low- and high-frequency domains were also determined. STZ induced similar levels of hyperglycemia and polydypsia in the groups. Mean arterial pressure (MAP) was increased from 100 +/- 6 to 124 +/- 6 mmHg in diabetic AT1aWT. MAP was unchanged in AT1aKO (80 +/- 4 vs. 85 +/- 5 mmHg, basal vs. STZ). Treatment with an ACE inhibitor, captopril, produced a greater reduction in MAP (-18%) in diabetic AT1aWT than in AT1aKO (-3.4%). BPV was lower in AT1aKO (19 +/- 0.5 vs. 9 +/- 2 mmHg(2), AT1aWT vs. AT1aKO). Diabetes reduced BPV but only in AT1aWT (19 +/- 0.5 vs. 8 +/- 1 mmHg(2), basal vs. STZ). There were no changes in HR in either group. In AT1aKO, STZ increased HRV and its high frequency domain with no changes seen in AT1aWT. Results document that ANG AT1a receptors are critical in diabetes-induced hypertension and in cardiac autonomic responses. PMID- 17122331 TI - Effects of cholecystokinin-8s in the nucleus tractus solitarius of vagally deafferented rats. AB - We have shown recently that cholecystokinin octapeptide (CCK-8s) increases glutamate release from nerve terminals onto neurons of the nucleus tractus solitarius pars centralis (cNTS). The effects of CCK on gastrointestinal-related functions have, however, been attributed almost exclusively to its paracrine action on vagal afferent fibers. Because it has been reported that systemic or perivagal capsaicin pretreatment abolishes the effects of CCK, the aim of the present work was to investigate the response of cNTS neurons to CCK-8s in vagally deafferented rats. In surgically deafferented rats, intraperitoneal administration of 1 or 3 mug/kg CCK-8s increased c-Fos expression in cNTS neurons (139 and 251% of control, respectively), suggesting that CCK-8s' effects are partially independent of vagal afferent fibers. Using whole cell patch-clamp techniques in thin brain stem slices, we observed that CCK-8s increased the frequency of spontaneous and miniature excitatory postsynaptic currents in 43% of the cNTS neurons via a presynaptic mechanism. In slices from deafferented rats, the percentage of cNTS neurons receiving glutamatergic inputs responding to CCK 8s decreased by approximately 50%, further suggesting that central terminals of vagal afferent fibers are not the sole site for the action of CCK-8s in the brain stem. Taken together, our data suggest that the sites of action of CCK-8s include the brain stem, and in cNTS, the actions of CCK-8s are not restricted to vagal central terminals but that nonvagal synapses are also involved. PMID- 17122332 TI - Mechanisms of blood pressure and heart rate variability: an insight from low level paraplegia. AB - It is still unclear whether the low-frequency oscillation in heart rate is generated by an endogenous neural oscillator or by a baroreflex resonance. Our aim was to investigate this issue by analyzing blood pressure and heart rate variability and the baroreflex function in paraplegic subjects with spinal cord injury below the fourth thoracic vertebra. These subjects were selected because they represent a model of intact central neural drive to the heart, with a partially impaired autonomic control of the vessels. In our study, arterial blood pressure and ECG were recorded in 33 able-bodied controls and in 33 subjects with spinal cord lesions between the fifth thoracic and the fourth lumbar vertebra 1) during supine rest (lowest sympathetic activation), 2) sitting on a wheelchair (light sympathetic activation), and 3) during exercise (moderate sympathetic activation). Blood pressure and heart rate spectra, coherence, and baroreflex function (sequence technique) were estimated in each condition. Compared with controls, paraplegic subjects showed a reduction of the low-frequency power of blood pressure and heart rate, and, unlike controls, a 0.1-Hz peak did not appear in their spectra. Sympathetic activation increased the 0.1-Hz peak of blood pressure and heart rate and the coherence at 0.1 Hz in controls only. Paraplegic subjects also had significantly lower baroreflex effectiveness and greater blood pressure variability. In conclusion, the disappearance of the 10-s oscillation of heart rate and blood pressure in subjects with spinal cord lesion supports the hypothesis of the baroreflex nature of this phenomenon. PMID- 17122333 TI - Intrinsic pump-conduit behavior of lymphangions. AB - Lymphangions, segments of lymphatic vessels bounded by valves, have characteristics of both ventricles and arteries. They can act primarily like pumps when actively transporting lymph against a pressure gradient. They also can act as conduit vessels when passively transporting lymph down a pressure gradient. This duality has implications for clinical treatment of several types of edema, since the strategy to optimize lymph flow may depend on whether it is most beneficial for lymphangions to act as pumps or conduits. To address this duality, we employed a simple computational model of a contracting lymphangion, predicted the flows at both positive and negative axial pressure gradients, and validated the results with in vitro experiments on bovine mesenteric vessels. This model illustrates that contraction increases flow for normal axial pressure gradients. With edema, limb elevation, or external compression, however, the pressure gradient might reverse, and lymph may flow passively down a pressure gradient. In such cases, the valves may be forced open during the entire contraction cycle. The vessel thus acts as a conduit, and contraction has the effect of increasing resistance to passive flow, thus inhibiting flow rather than promoting it. This analysis may explain a possible physiological benefit of the observed flow-mediated inhibition of the lymphatic pump at high flow rates. PMID- 17122334 TI - Increased 11beta-hydroxysteroid dehydrogenase type-1 and hexose-6-phosphate dehydrogenase in liver and adipose tissue of rat offspring exposed to alcohol in utero. AB - Rat offspring prenatally exposed to alcohol display features of metabolic syndrome characterized by a low birth weight, catch-up growth, dyslipidemia, and insulin-resistant diabetes with increased gluconeogenesis, during adult life. Gluconeogenesis is partly regulated by cyclic AMP- and glucocorticoid-dependent mechanisms. Glucocorticoid action at the receptor level depends on its circulating concentrations and is amplified at the prereceptor level by 11beta hydroxysteroid dehydrogenase type 1 (11beta-HSD1), which regenerates active glucocorticoids from inactive forms. To determine whether 11beta-HSD1 is dysregulated in this rat model, we examined the expression and enzyme activity of 11beta-HSD1 and its regulator enzyme hexose-6-phosphate dehydrogenase (H6PD) in the liver of postnatal day 7 (neonatal) and 3-mo-old (adult) rat offspring prenatally exposed to alcohol. Measurements of 11beta-HSD1 and H6PD were also performed in the omental fat of adult rat offspring. In both neonatal and adult rats, prenatal alcohol exposure resulted in increased tissue corticosterone concentrations, increased expression, and oxoreductase activity of 11beta-HSD1, and a parallel increase of H6PD expression. The data suggest that due to both transcriptional and posttranscriptional dysregulations, rats exposed to alcohol early in life have increased 11beta-HSD1 activity, which may explain insulin resistant diabetes in these animals later in life. PMID- 17122335 TI - Autoregulator protein PhaR for biosynthesis of polyhydroxybutyrate [P(3HB)] possibly has two separate domains that bind to the target DNA and P(3HB): Functional mapping of amino acid residues responsible for DNA binding. AB - PhaR from Paracoccus denitrificans functions as a repressor or autoregulator of the expression of genes encoding phasin protein (PhaP) and PhaR itself, both of which are components of polyhydroxyalkanoate (PHA) granules (A. Maehara, S. Taguchi, T. Nishiyama, T. Yamane, and Y. Doi, J. Bacteriol. 184:3992-4002, 2002). PhaR is a unique regulatory protein in that it also has the ability to bind tightly to an effector molecule, PHA polyester. In this study, by using a quartz crystal microbalance, we obtained direct evidence that PhaR binds to the target DNA and poly[(R)-3-hydroxybutyrate] [P(3HB)], one of the PHAs, at the same time. To identify the PhaR amino acid residues responsible for DNA binding, deletion and PCR-mediated random point mutation experiments were carried out with the gene encoding the PhaR protein. PhaR point mutants with decreased DNA-binding abilities were efficiently screened by an in vivo monitoring assay system coupled with gene expression of green fluorescent protein in Escherichia coli. DNA binding abilities of the wild-type and mutants of recombinant PhaR expressed in E. coli were evaluated using a gel shift assay and a surface plasmon resonance analysis. These experiments revealed that basic amino acids and a tyrosine in the N-terminal region, which is highly conserved among PhaR homologs, are responsible for DNA binding. However, most of the mutants with decreased DNA-binding abilities were unaffected in their ability to bind P(3HB), strongly suggesting that PhaR has two separate domains capable of binding to the target DNA and P(3HB). PMID- 17122336 TI - Genome-wide screening of genes required for swarming motility in Escherichia coli K-12. AB - Escherichia coli K-12 has the ability to migrate on semisolid media by means of swarming motility. A systematic and comprehensive collection of gene-disrupted E. coli K-12 mutants (the Keio collection) was used to identify the genes involved in the swarming motility of this bacterium. Of the 3,985 nonessential gene mutants, 294 were found to exhibit a strongly repressed-swarming phenotype. Further, 216 of the 294 mutants displayed no significant defects in swimming motility; therefore, the 216 genes were considered to be specifically associated with the swarming phenotype. The swarming-associated genes were classified into various functional categories, indicating that swarming is a specialized form of motility that requires a wide variety of cellular activities. These genes include genes for tricarboxylic acid cycle and glucose metabolism, iron acquisition, chaperones and protein-folding catalysts, signal transduction, and biosynthesis of cell surface components, such as lipopolysaccharide, the enterobacterial common antigen, and type 1 fimbriae. Lipopolysaccharide and the enterobacterial common antigen may be important surface-acting components that contribute to the reduction of surface tension, thereby facilitating the swarm migration in the E. coli K-12 strain. PMID- 17122337 TI - Role of GerD in germination of Bacillus subtilis spores. AB - Spores of a Bacillus subtilis strain with a gerD deletion mutation (Delta gerD) responded much slower than wild-type spores to nutrient germinants, although they did ultimately germinate, outgrow, and form colonies. Spores lacking GerD and nutrient germinant receptors also germinated slowly with nutrients, as did Delta gerD spores in which nutrient receptors were overexpressed. The germination defect of Delta gerD spores was not suppressed by many changes in the sporulation or germination conditions. Germination of Delta gerD spores was also slower than that of wild-type spores with a pressure of 150 MPa, which triggers spore germination through nutrient receptors. Ectopic expression of gerD suppressed the slow germination of Delta gerD spores with nutrients, but overexpression of GerD did not increase rates of spore germination. Loss of GerD had no effect on spore germination induced by agents that do not act through nutrient receptors, including a 1:1 chelate of Ca2+ and dipicolinic acid, dodecylamine, lysozyme in hypertonic medium, a pressure of 500 MPa, and spontaneous germination of spores that lack all nutrient receptors. Deletion of GerD's putative signal peptide or change of its likely diacylglycerylated cysteine residue to alanine reduced GerD function. The latter findings suggest that GerD is located in a spore membrane, most likely the inner membrane, where the nutrient receptors are located. All these data suggest that, while GerD is not essential for nutrient germination, this protein has an important role in spores' rapid response to nutrient germinants, by either direct interaction with nutrient receptors or some signal transduction essential for germination. PMID- 17122338 TI - Regulation of Vibrio polysaccharide synthesis and virulence factor production by CdgC, a GGDEF-EAL domain protein, in Vibrio cholerae. AB - In Vibrio cholerae, the second messenger 3',5'-cyclic diguanylic acid (c-di-GMP) regulates several cellular processes, such as formation of corrugated colony morphology, biofilm formation, motility, and virulence factor production. Both synthesis and degradation of c-di-GMP in the cell are modulated by proteins containing GGDEF and/or EAL domains, which function as a diguanylate cyclase and a phosphodiesterase, respectively. The expression of two genes, cdgC and mbaA, which encode proteins harboring both GGDEF and EAL domains is higher in the rugose phase variant of V. cholerae than in the smooth variant. In this study, we carried out gene expression analysis to determine the genes regulated by CdgC in the rugose and smooth phase variants of V. cholerae. We determined that CdgC regulates expression of genes required for V. cholerae polysaccharide synthesis and of the transcriptional regulator genes vpsR, vpsT, and hapR. CdgC also regulates expression of genes involved in extracellular protein secretion, flagellar biosynthesis, and virulence factor production. We then compared the genes regulated by CdgC and by MbaA, during both exponential and stationary phases of growth, to elucidate processes regulated by them. Identification of the regulons of CdgC and MbaA revealed that the regulons overlap, but the timing of regulation exerted by CdgC and MbaA is different, suggesting the interplay and complexity of the c-di-GMP signal transduction pathways operating in V. cholerae. PMID- 17122339 TI - The inner cavity of Escherichia coli DegP protein is not essential for molecular chaperone and proteolytic activity. AB - The Escherichia coli DegP protein is an essential periplasmic protein for bacterial survival at high temperatures. DegP has the unusual property of working as a chaperone below 28 degrees C, but efficiently degrading unfolded proteins above 28 degrees C. Monomeric DegP contains a protease domain and two PDZ domains. It oligomerizes into a hexameric cage through the staggered association of trimers. The active sites are located in a central cavity that is only accessible laterally, and the 12 PDZ domains act as mobile sidewalls that mediate opening and closing of the gates. As access to the active sites is restricted, DegP is an example of a self-compartmentalized protease. To determine the essential elements of DegP that maintain the integrity of the hexameric cage, we constructed several deletion mutants of DegP that formed trimers rather than hexamers. We found that residues 39 to 78 within the LA loops, as well as the PDZ2 domains are essential for the integrity of the DegP hexamer. In addition, we asked whether an enclosed cavity or cage of specific dimensions is required for the protease and chaperone activities in DegP. Both activities were maintained in the trimeric DegP mutants without an enclosed cavity and in deletion DegP mutants with significantly reduced dimensions of the cage. We conclude that the functional unit for the protease and chaperone activities of DegP is a trimer and that neither a cavity of specific dimensions nor the presence of an enclosed cavity appears to be essential for the protease and chaperone activities of DegP. PMID- 17122340 TI - Analysis of storage lipid accumulation in Alcanivorax borkumensis: Evidence for alternative triacylglycerol biosynthesis routes in bacteria. AB - Marine hydrocarbonoclastic bacteria, like Alcanivorax borkumensis, play a globally important role in bioremediation of petroleum oil contamination in marine ecosystems. Accumulation of storage lipids, serving as endogenous carbon and energy sources during starvation periods, might be a potential adaptation mechanism for coping with nutrient limitation, which is a frequent stress factor challenging those bacteria in their natural marine habitats. Here we report on the analysis of storage lipid biosynthesis in A. borkumensis strain SK2. Triacylglycerols (TAGs) and wax esters (WEs), but not poly(hydroxyalkanoic acids), are the principal storage lipids present in this and other hydrocarbonoclastic bacterial species. Although so far assumed to be a characteristic restricted to gram-positive actinomycetes, substantial accumulation of TAGs corresponding to a fatty acid content of more than 23% of the cellular dry weight is the first characteristic of large-scale de novo TAG biosynthesis in a gram-negative bacterium. The acyltransferase AtfA1 (ABO_2742) exhibiting wax ester synthase/acyl-coenzyme A:diacylglycerol acyltransferase (WS/DGAT) activity plays a key role in both TAG and WE biosynthesis, whereas AtfA2 (ABO_1804) was dispensable for storage lipid formation. However, reduced but still substantial residual TAG levels in atfA1 and atfA2 knockout mutants compellingly indicate the existence of a yet unknown WS/DGAT-independent alternative TAG biosynthesis route. Storage lipids of A. borkumensis were enriched in saturated fatty acids and accumulated as insoluble intracytoplasmic inclusions exhibiting great structural variety. Storage lipid accumulation provided only a slight growth advantage during short-term starvation periods but was not required for maintaining viability and long-term persistence during extended starvation phases. PMID- 17122341 TI - Membrane-spanning and periplasmic segments of CcmI have distinct functions during cytochrome c Biogenesis in Rhodobacter capsulatus. AB - In gram-negative bacteria, like Rhodobacter capsulatus, about 10 membrane-bound components (CcmABCDEFGHI and CcdA) are required for periplasmic maturation of c type cytochromes. These components perform the chaperoning and thio oxidoreduction of the apoproteins as well as the delivery and ligation of the heme cofactors. In the absence of any of these components, including CcmI, proposed to act as an apocytochrome c chaperone, R. capsulatus does not have the ability to produce holocytochromes c or consequently to exhibit photosynthetic growth and cytochrome cbb3 oxidase activity. Previously, we have demonstrated that null mutants of CcmI partially overcome cytochrome c deficiency phenotypes upon overproduction of the CcmF-R. capsulatus CcmH (CcmF-CcmH(Rc)) couple in a growth medium-dependent manner and fully bypass these defects by additional overproduction of CcmG. Here, we show that overproduction of the CcmF-CcmH(Rc) couple and overproduction of the N-terminal membrane-spanning segment of CcmI (CcmI-1) have similar suppression effects of cytochrome c maturation defects in CcmI-null mutants. Likewise, additional overproduction of CcmG, the C-terminal periplasmic segment of CcmI (CcmI-2), or even of apocytochrome c2 also provides complementation abilities similar to those of these mutants. These results indicate that the two segments of CcmI have different functions and support our earlier findings that two independent steps are required for full recovery of the loss of CcmI function. We therefore propose that CcmI-1 is part of the CcmF CcmH(Rc)-dependent heme ligation, while CcmI-2 is involved in the CcdA- and CcmG dependent apoprotein thioreduction steps, which intersect at the level of CcmI during cytochrome c biogenesis. PMID- 17122342 TI - Cyclohexa-1,5-diene-1-carbonyl-coenzyme A (CoA) hydratases of Geobacter metallireducens and Syntrophus aciditrophicus: Evidence for a common benzoyl-CoA degradation pathway in facultative and strict anaerobes. AB - In the denitrifying bacterium Thauera aromatica, the central intermediate of anaerobic aromatic metabolism, benzoyl-coenzyme A (CoA), is dearomatized by the ATP-dependent benzoyl-CoA reductase to cyclohexa-1,5-diene-1-carbonyl-CoA (dienoyl-CoA). The dienoyl-CoA is further metabolized by a series of beta oxidation-like reactions of the so-called benzoyl-CoA degradation pathway resulting in ring cleavage. Recently, evidence was obtained that obligately anaerobic bacteria that use aromatic growth substrates do not contain an ATP dependent benzoyl-CoA reductase. In these bacteria, the reactions involved in dearomatization and cleavage of the aromatic ring have not been shown, so far. In this work, a characteristic enzymatic step of the benzoyl-CoA pathway in obligate anaerobes was demonstrated and characterized. Dienoyl-CoA hydratase activities were determined in extracts of Geobacter metallireducens (iron reducing), Syntrophus aciditrophicus (fermenting), and Desulfococcus multivorans (sulfate reducing) cells grown with benzoate. The benzoate-induced genes putatively coding for the dienoyl-CoA hydratases in the benzoate degraders G. metallireducens and S. aciditrophicus were heterologously expressed and characterized. Both gene products specifically catalyzed the reversible hydration of dienoyl-CoA to 6 hydroxycyclohexenoyl-CoA (Km, 80 and 35 microM; Vmax, 350 and 550 micromol min( 1) mg(-1), respectively). Neither enzyme had significant activity with cyclohex-1 ene-1-carbonyl-CoA or crotonyl-CoA. The results suggest that benzoyl-CoA degradation proceeds via dienoyl-CoA and 6-hydroxycyclohexanoyl-CoA in strictly anaerobic bacteria. The steps involved in dienoyl-CoA metabolism appear identical in all nonphotosynthetic anaerobic bacteria, although totally different benzene ring-dearomatizing enzymes are present in facultative and obligate anaerobes. PMID- 17122343 TI - Novel type IV secretion system involved in propagation of genomic islands. AB - Type IV secretion systems (T4SSs) mediate horizontal gene transfer, thus contributing to genome plasticity, evolution of infectious pathogens, and dissemination of antibiotic resistance and other virulence traits. A gene cluster of the Haemophilus influenzae genomic island ICEHin1056 has been identified as a T4SS involved in the propagation of genomic islands. This T4SS is novel and evolutionarily distant from the previously described systems. Mutation analysis showed that inactivation of key genes of this system resulted in a loss of phenotypic traits provided by a T4SS. Seven of 10 mutants with a mutation in this T4SS did not express the type IV secretion pilus. Correspondingly, disruption of the genes resulted in up to 100,000-fold reductions in conjugation frequencies compared to those of the parent strain. Moreover, the expression of this T4SS was found to be positively regulated by one of its components, the tfc24 gene. We concluded that this gene cluster represents a novel family of T4SSs involved in propagation of genomic islands. PMID- 17122344 TI - Self-splicing of the bacteriophage T4 group I introns requires efficient translation of the pre-mRNA in vivo and correlates with the growth state of the infected bacterium. AB - Bacteriophage T4 contains three self-splicing group I introns in genes in de novo deoxyribonucleotide biosynthesis (in td, coding for thymidylate synthase and in nrdB and nrdD, coding for ribonucleotide reductase). Their presence in these genes has fueled speculations that the introns are retained within the phage genome due to a possible regulatory role in the control of de novo deoxyribonucleotide synthesis. To study whether sequences in the upstream exon interfere with proper intron folding and splicing, we inhibited translation in T4 infected bacteria as well as in bacteria containing recombinant plasmids carrying the nrdB intron. Splicing was strongly reduced for all three T4 introns after the addition of chloramphenicol during phage infection, suggesting that the need for translating ribosomes is a general trait for unperturbed splicing. The splicing of the cloned nrdB intron was markedly reduced in the presence of chloramphenicol or when translation was hindered by stop codons inserted in the upstream exon. Several exon regions capable of forming putative interactions with nrdB intron sequences were identified, and the removal or mutation of these exon regions restored splicing efficiency in the absence of translation. Interestingly, splicing of the cloned nrdB intron was also reduced as cells entered stationary phase and splicing of all three introns was reduced upon the T4 infection of stationary-phase bacteria. Our results imply that conditions likely to be frequently encountered by natural phage populations may limit the self-splicing efficiency of group I introns. This is the first time that environmental effects on bacterial growth have been linked to the regulation of splicing of phage introns. PMID- 17122345 TI - Lateral gene transfer in vitro in the intracellular pathogen Chlamydia trachomatis. AB - Genetic recombinants that resulted from lateral gene transfer (LGT) have been detected in sexually transmitted disease isolates of Chlamydia trachomatis, but a mechanism for LGT in C. trachomatis has not been described. We describe here a system that readily detects C. trachomatis LGT in vitro and that may facilitate discovery of its mechanisms. Host cells were simultaneously infected in the absence of antibiotics with an ofloxacin-resistant mutant and a second mutant that was resistant to lincomycin, trimethoprim, or rifampin. Selection for doubly resistant C. trachomatis isolates in the progeny detected apparent recombinant frequencies of 10(-4) to 10(-3), approximately 10(4) times more frequent than doubly resistant spontaneous mutants in progeny from uniparental control infections. Polyclonal doubly resistant populations and clones isolated from them in the absence of antibiotics had the specific resistance-conferring mutations present in the parental mutants; absence of the corresponding normal nucleotides indicated that they had been replaced by homologous recombination. These results eliminate spontaneous mutation, between-strain complementation, and heterotypic resistance as general explanations of multiply resistant C. trachomatis that originated in mixed infections in our experiments and demonstrate genetic stability of the recombinants. The kind of LGT we observed might be useful for creating new strains for functional studies by creating new alleles or combinations of alleles of polymorphic loci and might also disseminate antibiotic resistance genes in vivo. The apparent absence of phages and conjugative plasmids in C. trachomatis suggests that the LGT may have occurred by means of natural DNA transformation. Therefore, the experimental system may have implications for genetically altering C. trachomatis by means of DNA transfer. PMID- 17122346 TI - Crystal structure of uroporphyrinogen decarboxylase from Bacillus subtilis. AB - Uroporphyrinogen decarboxylase (UROD) is a branch point enzyme in the biosynthesis of the tetrapyrroles. It catalyzes the decarboxylation of four acetate groups of uroporphyrinogen III to yield coproporphyrinogen III, leading to heme and chlorophyll biosynthesis. UROD is a special type of nonoxidative decarboxylase, since no cofactor is essential for catalysis. In this work, the first crystal structure of a bacterial UROD, Bacillus subtilis UROD (UROD(Bs)), has been determined at a 2.3 A resolution. The biological unit of UROD(Bs) was determined by dynamic light scattering measurements to be a homodimer in solution. There are four molecules in the crystallographic asymmetric unit, corresponding to two homodimers. Structural comparison of UROD(Bs) with eukaryotic URODs reveals a variation of two loops, which possibly affect the binding of substrates and release of products. Structural comparison with the human UROD-coproporphyrinogen III complex discloses a similar active cleft, with five invariant polar residues (Arg29, Arg33, Asp78, Tyr154, and His322) and three invariant hydrophobic residues (Ile79, Phe144, and Phe207), in UROD(Bs). Among them, Asp78 may interact with the pyrrole NH groups of the substrate, and Arg29 is a candidate for positioning the acetate groups of the substrate. Both residues may also play catalytic roles. PMID- 17122347 TI - Structural characterization of a specific glycopeptidolipid containing a novel N acyl-deoxy sugar from mycobacterium intracellulare serotype 7 and genetic analysis of its glycosylation pathway. AB - The nontuberculous Mycobacterium avium-Mycobacterium intracellulare complex (MAC) is distributed ubiquitously in the environment and is an important cause of respiratory and lymphatic disease in humans and animals. These species produce polar glycopeptidolipids (GPLs), and of particular interest is their serotype specific antigenicity. Structurally, GPLs contain an N-acylated tetrapeptide amino alcohol core that is glycosylated at the C terminal with 3,4-di-O-methyl rhamnose and at the d-allo-threonine with a 6-deoxy-talose. This serotype nonspecific GPL is found in all MAC species. The serotype-specific GPLs are further glycosylated with a variable haptenic oligosaccharide at 6-deoxy-talose. At present, 31 distinct serotype-specific GPLs have been identified on the basis of oligosaccharide composition, and the complete structures of 14 serotype specific GPLs have been defined. It is considered that the modification of the GPL structure plays an important role in bacterial physiology, pathogenesis, and host immune responses. In this study, we defined the complete structure of a novel serotype 7 GPL that has a unique terminal amido sugar. The main molecular mass is 1,874, and attached to the tetrapeptide-amino alcohol core is the serotype 7-specific oligosaccharide unit of 4-2'-hydroxypropanoyl-amido-4,6 dideoxy-2-O-methyl-beta-hexose-(1-->3)-alpha-l-rhamnose-(1-->3)-alpha-l-rhamnose (1-->3)-alpha-l-rhamnose-(1-->2)-alpha-l-6-deoxy-talose. Moreover, we isolated and characterized the serotype 7-specific gene cluster involved in glycosylation of the oligosaccharide. Nine open reading frames (ORFs) were observed in the cluster. Based on the sequence homology, the ORFs are thought to participate in the biosynthesis of the serotype 7 GPL. PMID- 17122348 TI - Regulation of denitrification genes in Neisseria meningitidis by nitric oxide and the repressor NsrR. AB - The human pathogen Neisseria meningitidis is capable of growth using the denitrification of nitrite to nitrous oxide under microaerobic conditions. This process is catalyzed by two reductases: nitrite reductase (encoded by aniA) and nitric oxide (NO) reductase (encoded by norB). Here, we show that in N. meningitidis MC58 norB is regulated by nitric oxide via the product of gene NMB0437 which encodes NsrR. NsrR is a repressor in the absence of NO, but norB expression is derepressed by NO in an NsrR-dependent manner. nsrR-deficient mutants grow by denitrification more rapidly than wild-type N. meningitidis, and this is coincident with the upregulation of both NO reductase and nitrite reductase even under aerobic conditions in the absence of nitrite or NO. The NsrR dependent repression of aniA (unlike that of norB) is not lifted in the presence of NO. The role of NsrR in the control of expression of aniA is linked to the function of the anaerobic activator protein FNR: analysis of nsrR and fnr single and nsrR fnr double mutants carrying an aniA promoter lacZ fusion indicates that the role of NsrR is to prevent FNR-dependent aniA expression under aerobic conditions, indicating that FNR in N. meningitidis retains considerable activity aerobically. PMID- 17122349 TI - Integration and excision of a newly discovered bacteroides conjugative transposon, CTnBST. AB - Conjugative transposons (CTns) are major contributors to the spread of antibiotic resistance genes among Bacteroides species. CTnBST, a newly discovered Bacteroides conjugative transposon, carries an erythromycin resistance gene, ermB, and previously has been estimated to be about 100 kbp in size. We report here the locations and sequencing of both of its ends. We have also located and sequenced the gene that catalyzes the integration of CTnBST, intBST. The integrase gene encodes a 377-amino-acid protein that has the C-terminal R-K-H-R-H Y motif that is characteristic of members of the tyrosine recombinase family of integrases. DNA sequence comparisons of the ends of CTnBST, the joined ends of the circular intermediate, and the preferred site into which the circular form of CTnBST had integrated revealed that the preferred integration site (attB1) contained an 18-bp sequence of identity to the crossover region, attBST, on CTnBST. Although this site was used in about one-half of the integration events, sequence analysis of these integration events revealed that both CTnBST and a miniature form of CTnBST (miniBST) integrated into a variety of other sites in the chromosome. All of the sites had two conserved regions, AATCTG and AAAT. These two regions flanked a 2-bp sequence, bp 10 and bp 11 of the 18-bp sequence, that varied in some of the different sites and sometimes in the attBST sequences. Our results suggest that CTnBST integrates site selectively and that the crossover appears to occur within a 12-bp region that contains the two regions of conserved sequences. PMID- 17122350 TI - Phosphoinositides are involved in control of the glucose-dependent growth resumption that follows the transition phase in Streptomyces lividans. AB - The interruption of the sblA gene of Streptomyces lividans was previously shown to lead to relief of glucose repression of the normally strongly glucose repressed alpha-amylase gene. In addition to this relief, an early entry into stationary phase was observed when cells were grown in a minimal medium containing glucose as the main carbon source. In this study, we established that this mutant does not resume growth after the transition phase when cultured in the complex glucose-rich liquid medium R2YE and sporulates much earlier than the wild-type strain when plated on solid R2YE. These phenotypic differences, which were abolished when glucose was omitted from the R2YE medium, correlated with a reduced glucose uptake ability of the sblA mutant strain. sblA was shown to encode a bifunctional enzyme possessing phospholipase C-like and phosphoinositide phosphatase activities. The cleavage of phosphoinositides by SblA seems necessary to trigger the glucose-dependent renewed growth that follows the transition phase. The transient expression of sblA that takes place just before the transition phase is consistent with a regulatory role for this gene during the late stages of growth. The tight temporal control of sblA expression was shown to depend on two operator sites. One, located just upstream of the -35 promoter region, likely constitutes a repressor binding site. The other, located 170 bp downstream of the GTG sblA translational start codon, may be involved in the regulation of the degradation of the sblA transcript. This study suggests that phosphoinositides constitute important regulatory molecules in Streptomyces, as they do in eukaryotes. PMID- 17122351 TI - Identification of genes regulated by the cepIR quorum-sensing system in Burkholderia cenocepacia by high-throughput screening of a random promoter library. AB - The Burkholderia cenocepacia cepIR quorum-sensing system regulates expression of extracellular proteases, chitinase, and genes involved in ornibactin biosynthesis, biofilm formation, and motility. In a genome-wide screen we identified cepIR-regulated genes by screening a random promoter library of B. cenocepacia K56-2 constructed in a luminescence reporter detection plasmid for differential expression in response to N-octanoyl-l-homoserine lactone (OHL). Eighty-nine clones were identified; in 58 of these clones expression was positively regulated by cepIR, and in 31 expression was negatively regulated by cepIR. The expression profiles of the 89 promoter clones were compared in the cepI mutant K56-dI2 in medium supplemented with 30 pM OHL and K56-2 to confirm that the presence of OHL restored expression to wild-type levels. To validate the promoter library observations and to determine the effect of a cepR mutation on expression of selected genes, the mRNA levels of nine genes whose promoters were predicted to be regulated by cepR were quantitated by quantitative reverse transcription-PCR in the wild type and cepI and cepR mutants. The expression levels of all nine genes were similar in the cepI and cepR mutants and consistent with the promoter-lux reporter activity. The expression of four selected cepIR regulated gene promoters was examined in a cciIR mutant, and two of these promoters were also regulated by cciIR. This study extends our understanding of genes whose expression is influenced by cepIR and indicates the global regulatory effect of the cepIR system in B. cenocepacia. PMID- 17122352 TI - Involvement of the cynABDS operon and the CO2-concentrating mechanism in the light-dependent transport and metabolism of cyanate by cyanobacteria. AB - The cyanobacteria Synechococcus elongatus strain PCC7942 and Synechococcus sp. strain UTEX625 decomposed exogenously supplied cyanate (NCO-) to CO2 and NH3 through the action of a cytosolic cyanase which required HCO3- as a second substrate. The ability to metabolize NCO- relied on three essential elements: proteins encoded by the cynABDS operon, the biophysical activity of the CO2 concentrating mechanism (CCM), and light. Inactivation of cynS, encoding cyanase, and cynA yielded mutants unable to decompose cyanate. Furthermore, loss of CynA, the periplasmic binding protein of a multicomponent ABC-type transporter, resulted in loss of active cyanate transport. Competition experiments revealed that native transport systems for CO2, HCO3-, NO3-, NO2-, Cl-, PO4(2-), and SO4(2 ) did not contribute to the cellular flux of NCO- and that CynABD did not contribute to the flux of these nutrients, implicating CynABD as a novel primary active NCO- transporter. In the S. elongatus strain PCC7942 DeltachpX DeltachpY mutant that is defective in the full expression of the CCM, mass spectrometry revealed that the cellular rate of cyanate decomposition depended upon the size of the internal inorganic carbon (Ci) (HCO3- + CO2) pool. Unlike wild-type cells, the rate of NCO- decomposition by the DeltachpX DeltachpY mutant was severely depressed at low external Ci concentrations, indicating that the CCM was essential in providing HCO3- for cyanase under typical growth conditions. Light was required to activate and/or energize the active transport of both NCO- and Ci. Putative cynABDS operons were identified in the genomes of diverse Proteobacteria, suggesting that CynABDS-mediated cyanate metabolism is not restricted to cyanobacteria. PMID- 17122353 TI - Paraoxonase-2 deficiency enhances Pseudomonas aeruginosa quorum sensing in murine tracheal epithelia. AB - Pseudomonas aeruginosa is an important cause of nosocomial infections and is frequently present in the airways of cystic fibrosis patients. Quorum sensing mediates P. aeruginosa's virulence and biofilm formation through density dependent interbacterial signaling with autoinducers. N-3-oxododecanoyl homoserine lactone (3OC12-HSL) is the major autoinducer in P. aeruginosa. We have previously shown that human airway epithelia and paraoxonases (PONs) degrade 3OC12-HSL. This study investigated the role of PON1, PON2, and PON3 in airway epithelial cell inactivation of 3OC12-HSL. All three PONs were present in murine tracheal epithelial cells, with PON2 and PON3 expressed at the highest levels. Lysates of tracheal epithelial cells from PON2, but not PON1 or PON3, knockout mice had impaired 3OC12-HSL inactivation compared with wild-type mice. In contrast, PON1-, PON2-, or PON3-targeted deletions did not affect 3OC12-HSL degradation by intact epithelia. Overexpression of PON2 enhanced 3OC12-HSL degradation by human airway epithelial cell lysates but not by intact epithelia. Finally, using a quorum-sensing reporter strain of P. aeruginosa, we found that quorum sensing was enhanced in PON2-deficient airway epithelia. In summary, these results show that loss of PON2 impairs 3OC12-HSL degradation by airway epithelial cells and suggests that diffusion of 3OC12-HSL into the airway cells can be the rate-limiting step for degradation of the molecule. PMID- 17122354 TI - Regulation of lipopolysaccharide-induced increases in neutrophil glucose uptake. AB - The pathogenesis of many lung diseases involves neutrophilic inflammation. Neutrophil functions, in turn, are critically dependent on glucose uptake and glycolysis to supply the necessary energy to meet these functions. In this study, we determined the effects of p38 mitogen-activated protein kinase and hypoxia inducible factor (HIF)-1, as well as their potential interaction, on the expression of membrane glucose transporters and on glucose uptake in murine neutrophils. Neutrophils were harvested and purified from C57BL/6 mice and stimulated with lipopolysaccharide (LPS) in the presence or absence of specific p38 and HIF-1 inhibitors. Glucose uptake was measured as the rate of [3H]deoxyglucose (DG) uptake. We identified GLUT-1 in mouse neutrophils, but neither GLUT-3 nor GLUT-4 were detected using Western blot analysis, even after LPS stimulation. LPS stimulation did not increase GLUT-1 protein levels but did cause translocation of GLUT-1 from the cell interior to the cell surface, together with a dose-dependent increase in [3H]DG uptake, indicating that glucose uptake is regulated in these cells. LPS also activated both p38 and the HIF-1 pathway. Inhibitors of p38 and HIF-1 blocked GLUT-1 translocation and [3H]DG uptake. These data suggest that LPS-induced increases in neutrophil glucose uptake are mediated by GLUT-1 translocation to the cell surface in response to sequential activation of neutrophil p38 and HIF-1alpha in neutrophils. Given that neutrophil function and glucose metabolism are closely linked, control of the latter may represent a new target to ameliorate the deleterious effects of neutrophils on the lungs. PMID- 17122355 TI - Glutathione availability modulates alveolar macrophage function in the chronic ethanol-fed rat. AB - We have previously demonstrated that chronic alcohol exposure decreases glutathione in the alveolar space. Although alcohol use is associated with decreased alveolar macrophage function, the mechanism by which alcohol impairs macrophage phagocytosis is unknown. In the current study, we examined the possibility that ethanol-induced alveolar macrophage dysfunction was secondary to decreased glutathione and subsequent chronic oxidative stress in the alveolar space. After 6 wk of ethanol ingestion, oxidant stress in the alveolar macrophages was evidenced by a 30-mV oxidation of the GSH/GSSG redox potential (P or =100 microM) reduced the compaction rate of BATs in a dose-dependent manner. ADP, 2'-(or 3')-O-(4 benzoylbenzoyl) ATP, and UTP, but not alpha,beta-methylene ATP, also reduced the compaction rate but to a lesser extent. Pyridoxal-phosphate-6-azophenyl-2',4' disulfonic acid tetrasodium did not block the effect of ATP on BAT compaction rate. These results indicate that both P2X and P2Y receptors are involved in ATP induced reduction of BAT compaction rate. Steady fluid flow and RT-PCR results showed that ATP reduced cell attachment on type I collagen by downregulating the expression of integrin alpha(1). These results suggest a potential role for P2 receptors in matrix remodeling and repair and as a potential drug target in treatment of bone diseases. PMID- 17122381 TI - Muscle size, neuromuscular activation, and rapid force characteristics in elderly men and women: effects of unilateral long-term disuse due to hip-osteoarthritis. AB - Substantial evidence exists for the age-related decline in muscle strength and neural function, but the effect of long-term disuse in the elderly is largely unexplored. The present study examined the effect of unilateral long-term limb disuse on maximal voluntary quadriceps contraction (MVC), lean quadriceps muscle cross-sectional area (LCSA), contractile rate of force development (RFD, Delta force/Delta time), impulse (integral force dt), muscle activation deficit (interpolated twitch technique), maximal neuromuscular activity [electromyogram (EMG)], and antagonist muscle coactivation in elderly men (M: 60-86 yr; n = 19) and women (W: 60-86 yr; n = 20) with unilateral chronic hip-osteoarthritis. Both sides were examined to compare the effect of long-term decreased activity on the affected (AF) leg with the unaffected (UN) side. AF had a significant lower MVC (W: 20%; M: 20%), LCSA (W: 8%; M: 10%), contractile RFD (W: 17-26%; M: 15-24%), impulse (W: 10-19%, M: 19-20%), maximal EMG amplitude (W: 22-25%, M: 22-28%), and an increased muscle activation deficit (-18%) compared with UN. Furthermore, women were less strong (AF: 40%; UN: 39%), had less muscle mass (AF: 33%; UN: 34%), and had a lower RFD (AF: 38-50%; UN: 41-48%) compared with men. Similarly, maximum EMG amplitude was smaller for both agonists (AF: 51-63%; UN: 35-61%) and antagonist (AF: 49-64%; UN: 36-56%) muscles in women compared with men. However, when MVC and RFD were normalized to LCSA, there were no differences between genders. The present data demonstrate that disuse leads to a marked loss of muscle strength and muscle mass in elderly individuals. Furthermore, the data indicate that neuromuscular activation and contractile RFD are more affected by long-term disuse than maximal muscle strength, which may increase the future risk for falls. PMID- 17122382 TI - Opiate therapy in chronic cough. AB - RATIONALE: Cough is the most common complaint for which medical attention is sought, and chronic cough can be both physically and mentally debilitating. There is currently no evidence supporting the use of antitussives in chronic treatment resistant cough. OBJECTIVES: We tested the hypothesis that morphine sulfate in the dose of 5 mg twice daily would bring about a reduction in cough frequency and severity in patients failing to respond to specific measures. METHODS: Patients recruited from the Hull Cough Clinic were enrolled into a randomized double-blind placebo-controlled study using 4 weeks of slow-release morphine sulfate and a corresponding period of matched placebo. An open-labeled extension of the core study allowed dose escalation to 10 mg twice daily. Cough was assessed using the Leicester Cough Questionnaire, daily symptom diary, and citric acid cough challenge. RESULTS: Twenty-seven patients completed the core study. A significant improvement of 3.2 points over baseline was noted on the Leicester Cough Questionnaire (p < 0.01). A rapid and highly significant reduction by 40% in daily cough scores was noted among patients on slow-release morphine sulfate (p < 0.01). Objective testing of the cough reflex using citric acid cough challenge tests did not show any significant changes. Eighteen patients continued into the extension study. Two-thirds of these patients opted to increase the morphine to 10 mg twice daily. At the end of 3 months, there was a similar improvement in cough between the 5- and 10-mg groups. CONCLUSION: Morphine sulfate is an effective antitussive in intractable chronic cough at the doses of 5 to 10 mg twice daily. PMID- 17122383 TI - Helminth-derived products inhibit the development of allergic responses in mice. AB - RATIONALE: Epidemiological studies suggest that infections with helminths protect from the development of asthma. Supporting this view is our published finding that infection with Nippostrongylus brasiliensis decreased ovalbumin-induced Th2 responses in the lung of mice. OBJECTIVES: To evaluate if N. brasiliensis excretory-secretory products also prevent the development of asthma. METHODS: Mice were immunized with ovalbumin/alum intraperitoneally in the absence or presence of helminthic products and then challenged intranasally with ovalbumin. Six days later, we analyzed if the mice developed Th2 responses in the lung. MAIN RESULTS: The application of the helminthic products together with ovalbumin/alum during the sensitization period totally inhibited the development of eosinophilia and goblet cell metaplasia in the airways and also strongly reduced the development of airway hyperreactivity. Allergen-specific IgG1 and IgE serum levels were also strongly reduced. These findings correlated with decreased levels of IL-4 and IL-5 in the airways in product-treated animals. The suppressive effects on the development of allergic responses were independent of the presence of Toll-like receptors 2 and 4, IFN-gamma, and most important, IL 10. Interestingly, suppression was still observed when the helminthic products were heated or treated with proteinase K. Paradoxically, we found that strong helminth product-specific Th2 responses were induced in parallel with the inhibition of ovalbumin-specific responses. CONCLUSION: Our results suggest that helminths suppress the development of asthma by secreting substances that modulate allergic responses without affecting the generation of helminth-specific Th2 immunity. The identification of these products may lead to the design of novel therapeutic intervention strategies for the treatment of asthma. PMID- 17122384 TI - Interaction of the Ca2+-sensing receptor with the inwardly rectifying potassium channels Kir4.1 and Kir4.2 results in inhibition of channel function. AB - The Ca(2+)-sensing receptor (CaR), a G protein-coupled receptor, is expressed in many epithelial tissues including the parathyroid glands, kidney, and GI tract. Although its role in regulating PTH levels and Ca(2+) metabolism are best characterized, it may also regulate salt and water transport in the kidney as demonstrated by recent reports showing association of potent gain-of-function mutations in the CaR with a Bartter-like, salt-wasting phenotype. To determine whether this receptor interacts with novel proteins that control ion transport, we screened a human adult kidney cDNA library with the COOH-terminal 219 amino acid cytoplasmic tail of the CaR as bait using the yeast two-hybrid system. We identified two independent clones coding for approximately 125 aa from the COOH terminus of the inwardly rectifying K(+) channel, Kir4.2. The CaR and Kir4.2 as well as Kir4.1 (another member of Kir4 subfamily) were reciprocally coimmunoprecipitated from HEK-293 cells in which they were expressed, but the receptor did not coimmunoprecipitate with Kir5.1 or Kir1.1. Both Kir4.1 and Kir4.2 were immunoprecipitated from rat kidney extracts with the CaR. In Xenopus laevis oocytes, expression of the CaR with either Kir4.1 or Kir4.2 channels resulted in inactivation of whole cell current as measured by two-electrode voltage clamp, but the nonfunctional CaR mutant CaR(R796W), and that does not coimmunoprecipitate with the channels, had no effect. Kir4.1 and the CaR were colocalized in the basolateral membrane of the distal nephron. The CaR interacts directly with Kir4.1 and Kir4.2 and can decrease their currents, which in turn could reduce recycling of K(+) for the basolateral Na(+)-K(+)-ATPase and thereby contribute to inhibition of Na(+) reabsorption. PMID- 17122385 TI - Acute unilateral ureteral distension inhibits glutamate-dependent spinal pelvic urethra reflex potentiation via GABAergic neurotransmission in anesthetized rats. AB - The effects of an acute increase in intraureteral pressure (IUP) on pelvic urethra reflex potentiation were examined in urethane-anesthetized rats by recording the external urethral sphincter electromyogram activities evoked by the pelvic afferent stimulation. Compared with a single action potential elicited by the test stimulation (TS; characterized by an intensity that evoked a constant reflex response without facilitation, 1/30 Hz, 1.03 +/- 0.12 spikes/stimulation, n = 7), the repetitive stimulation [RS; identical stimulation intensity as the TS (1 Hz)] significantly induced spinal reflex potentiation (SRP; 16.90 +/- 2.00 spikes/stimulation, P < 0.01, n = 7). Such SRP was significantly attenuated by intrathecal 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo (F) quinoxaline [NBQX; a glutamatergic alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionat (AMPA) receptor antagonist] and d-2-amino-5-phosphonovalerate [APV; a glutamatergic N methyl-d-aspartate (NMDA) antagonist; the spike number per stimulation: 11.0 +/- 0.70 for NBQX, 1.01 +/- 0.30 for APV, and 16.90 +/- 2.0 for RS, respectively, n = 7, P < 0.01]. Acute stepwise elevations of IUP gradually attenuated and eventually abolished the RS-induced SRP (16.80 +/- 1.30, 17.00 +/- 1.30, 16.30 +/ 1.30, 10.50 +/- 1.80, 8.80 +/- 1.90, 3.50 +/- 1.60, 0.80 +/- 0.20, 0.70 +/- 0.20, and 0.20 +/- 0.10 spikes/stimulation at intraureteral pressure of 0, 2.5, 5, 7.5, 10, 12.5, 15, 17.5, and 20 cmH(2)O, respectively, n = 7). Intrathecal NMDA (a glutamatergic NMDA receptor agonist) and bicuculline (a GABA receptor antagonist) both reversed the abolition of RS-induced SRP caused by unilateral ureteral distension (14.0 +/- 4.04 and 8.00 +/- 1.53 spikes/stimulation, respectively, n = 7, P < 0.01). All the results suggested unilateral ureteral distension might compensatorily relax the urethra via GABAergic inhibition of NMDA-dependent SRP. PMID- 17122386 TI - Effect of renal injury-induced neurogenic hypertension on NO synthase, caveolin 1, AKt, calmodulin and soluble guanylate cyclase expressions in the kidney. AB - Single injection of a small quantity of phenol into the cortex of one kidney in rats results in development of persistent hypertension (HTN) which is thought to be mediated by activation of renal afferent and efferent sympathetic pathways and sodium retention. Nitric oxide (NO) plays a major role in regulation of renal vascular resistance, tubular Na(+) reabsorption, pressure natriuresis, and thereby systemic arterial pressure. The present study was performed to test the hypothesis that chronic renal injury-induced HTN may be associated with dysregulation of NO system in the kidney. Accordingly, urinary NO metabolite (NO(x)) and cGMP excretions as well as renal cortical tissue (right kidney) expressions of NO synthase (NOS) isoforms [endothelial, neuronal, and inducible NOS, respectively (eNOS, nNOS, and iNOS)], NOS-regulatory factors (Caveolin-1, phospho-AKt, and calmodulin), and second-messenger system (soluble guanylate cyclase [sGC] and phosphodiesterase-5 [PDE-5]) were determined in male Sprague Dawley rats 4 wk after injection of phenol (50 mul of 10% phenol) or saline into the lower pole of left kidney. The phenol-injected group exhibited a significant elevation of arterial pressure, marked reductions of urinary NO(x) and cGMP excretions, downregulations of renal tissue nNOS, eNOS, Phospho-eNOS, iNOS, and alpha chain of sGC. However, renal tissue AKt, phospho-AKT, Calmodulin, and PDE-5 proteins were unchanged in the phenol-injected animals. In conclusion, renal injury in this model results in significant downregulations of NOS isoforms and sGC and consequent reductions of NO production and cGMP generation by the kidney, events that may contribute to maintenance of HTN in this model. PMID- 17122387 TI - Limited functional redundancy and oscillation of cyclins in multinucleated Ashbya gossypii fungal cells. AB - Cyclin protein behavior has not been systematically investigated in multinucleated cells with asynchronous mitoses. Cyclins are canonical oscillating cell cycle proteins, but it is unclear how fluctuating protein gradients can be established in multinucleated cells where nuclei in different stages of the division cycle share the cytoplasm. Previous work in A. gossypii, a filamentous fungus in which nuclei divide asynchronously in a common cytoplasm, demonstrated that one G1 and one B-type cyclin do not fluctuate in abundance across the division cycle. We have undertaken a comprehensive analysis of all G1 and B-type cyclins in A. gossypii to determine whether any of the cyclins show periodic abundance across the cell cycle and to examine whether cyclins exhibit functional redundancy in such a cellular environment. We localized all G1 and B-type cyclins and notably found that only AgClb5/6p varies in subcellular localization during the division cycle. AgClb5/6p is lost from nuclei at the meta-anaphase transition in a D-box-dependent manner. These data demonstrate that efficient nuclear autonomous protein degradation can occur within multinucleated cells residing in a common cytoplasm. We have shown that three of the five cyclins in A. gossypii are essential genes, indicating that there is minimal functional redundancy in this multinucleated system. In addition, we have identified a cyclin, AgClb3/4p, that is essential only for sporulation. We propose that the cohabitation of different cyclins in nuclei has led to enhanced substrate specificity and limited functional redundancy within classes of cyclins in multinucleated cells. PMID- 17122388 TI - Antibiotic resistances of starter and probiotic strains of lactic acid bacteria. AB - The antibiotic resistances of 45 lactic acid bacteria strains belonging to the genera Lactobacillus, Streptococcus, Lactococcus, Pediococcus, and Leuconostoc were investigated. The objective was to determine antibiotic resistances and to verify these at the genetic level, as is currently suggested by the European "qualified presumption of safety" safety evaluation system for industrial starter strains. In addition, we sought to pinpoint possible problems in resistance determinations. Primers were used to PCR amplify genes involved in beta-lactam antibiotic, chloramphenicol, tetracycline, and erythromycin resistance. The presence of ribosomal protection protein genes and the ermB gene was also determined by using a gene probe. Generally, the incidences of erythromycin, chloramphenicol, tetracycline, or beta-lactam resistances in this study were low (<7%). In contrast, aminoglycoside (gentamicin and streptomycin) and ciprofloxacin resistances were higher than 70%, indicating that these may constitute intrinsic resistances. The genetic basis for ciprofloxacin resistance could not be verified, since no mutations typical of quinolone resistances were detected in the quinolone determining regions of the parC and gyrA genes. Some starter strains showed low-level ampicillin, penicillin, chloramphenicol, and tetracycline resistances, but no known resistance genes could be detected. Although some strains possessed the cat gene, none of these were phenotypically resistant to chloramphenicol. Using reverse transcription-PCR, these cat genes were shown to be silent under both inducing and noninducing conditions. Only Lactobacillus salivarius BFE 7441 possessed an ermB gene, which was encoded on the chromosome and which could not be transferred in filter-mating experiments. This study clearly demonstrates problems encountered with resistance testing, in that the breakpoint values are often inadequately identified, resistance genes may be present but silent, and the genetic basis and associated resistance mechanisms toward some antibiotics are still unknown. PMID- 17122389 TI - Operon prediction for sequenced bacterial genomes without experimental information. AB - Various computational approaches have been proposed for operon prediction, but most algorithms rely on experimental or functional data that are only available for a small subset of sequenced genomes. In this study, we explored the possibility of using phylogenetic information to aid in operon prediction, and we constructed a Bayesian hidden Markov model that incorporates comparative genomic data with traditional predictors, such as intergenic distances. The prediction algorithm performs as well as the best previously reported method, with several significant advantages. It uses fewer data sources and so it is easier to implement, and the method is more broadly applicable than previous methods--it can be applied to essentially every gene in any sequenced bacterial genome. Furthermore, we show that near-optimal performance is easily reached with a generic set of comparative genomes and does not depend on a specific relationship between the subject genome and the comparative set. We applied the algorithm to the Bacillus anthracis genome and found that it successfully predicted all previously verified B. anthracis operons. To further test its performance, we chose a predicted operon (BA1489-92) containing several genes with little apparent functional relatedness and tested their cotranscriptional nature. Experimental evidence shows that these genes are cotranscribed, and the data have interesting implications for B. anthracis biology. Overall, our findings show that this algorithm is capable of highly sensitive and accurate operon prediction in a wide range of bacterial genomes and that these predictions can lead to the rapid discovery of new functional relationships among genes. PMID- 17122390 TI - Evaluation of a wipe surface sample method for collection of Bacillus spores from nonporous surfaces. AB - Polyester-rayon blend wipes were evaluated for efficiency of extraction and recovery of powdered Bacillus atrophaeus spores from stainless steel and painted wallboard surfaces. Method limits of detection were also estimated for both surfaces. The observed mean efficiency of polyester-rayon blend wipe recovery from stainless steel was 0.35 with a standard deviation of +/-0.12, and for painted wallboard it was 0.29 with a standard deviation of +/-0.15. Evaluation of a sonication extraction method for the polyester-rayon blend wipes produced a mean extraction efficiency of 0.93 with a standard deviation of +/-0.09. Wipe recovery quantitative limits of detection were estimated at 90 CFU per unit of stainless steel sample area and 105 CFU per unit of painted wallboard sample area. The method recovery efficiency and limits of detection established in this work provide useful guidance for the planning of incident response environmental sampling following the release of a biological agent such as Bacillus anthracis. PMID- 17122391 TI - Exopolysaccharide expression in Lactococcus lactis subsp. cremoris Ropy352: evidence for novel gene organization. AB - Lactococcus lactis subsp. cremoris Ropy352 produces two distinct heteropolysaccharides, phenotypically described as ropy and mucoid, when cultured in nonfat milk. One exopolysaccharide precipitated with 50% ethanol as a series of elongated threads and was composed of glucose and galactose in a molar ratio of 3:2. The second exopolysaccharide precipitated with 75% ethanol as a fine flocculant and consisted of galactose, glucose, and mannose with a molar ratio of 67:21:12. A mutant strain, L. lactis subsp. cremoris EK240, lacking the ropy phenotype did not produce the exopolysaccharide that precipitated with 50% ethanol; however, it produced the exopolysaccharide that precipitated with 75% ethanol, indicating that the former exopolysaccharide is essential for the ropy phenotype. Cultures of L. lactis subsp. cremoris Ropy352 in 10% nonfat milk reached a viscosity of 25 Pa-s after 24 h, while those of the nonropy L. lactis subsp. cremoris EK240 mutant did not change. A mutation abolishing ropy exopolysaccharide expression mapped to a region on a plasmid containing two open reading frames, epsM and epsN, encoding novel glycosyltransferases bordered by ISS1 elements oriented in the same direction. Sequencing of this plasmid revealed two other regions involved in exopolysaccharide expression, an operon located between partial IS981 and IS982 elements, and an independent gene, epsU. Two and possibly three of these regions are involved in L. lactis subsp. cremoris Ropy352 exopolysaccharide expression and are arranged in a novel fashion different from that of typical lactococcal exopolysaccharide loci, and this provides genetic evidence for exopolysaccharide gene reorganization and evolution in Lactococcus. PMID- 17122392 TI - Inhibitory impact of bifidobacteria on the transfer of beta-lactam resistance among Enterobacteriaceae in the gnotobiotic mouse digestive tract. AB - While looking for new means to limit the dissemination of antibiotic resistance, we evaluated the role of potentially probiotic bifidobacteria on the transfer of resistance genes between enterobacteria. Transfers of bla genes encoding extended spectrum beta-lactamases (SHV-5 and CTX-M-15) were studied in the absence or presence of bifidobacteria. In vitro, transfer frequencies of these bla genes decreased significantly in the presence of three of five tested strains, i.e., Bifidobacterium longum CUETM-89-215, Bifidobacterium bifidum CIP-56.7T, and Bifidobacterium pseudocatenulatum CIP-104168T. Four transfer experiments were conducted in the digestive tract of gnotobiotic mice, the first three observing the effect of B. longum CUETM-89-215, B. bifidum CIP-56.7T, and B. pseudocatenulatum CIP-104168T on blaSHV-5 transfer and the fourth experiment studying the effect of B. bifidum CIP-56.7T on blaCTX-M-15 transfer. These experiments revealed significant decreases in the transconjugant levels (up to 3 logs) in mice having received B. bifidum CIP-56.7T or B. pseudocatenulatum CIP 104168T compared to control mice. Bifidobacteria appear to have an inhibitory impact on the transfer of antibiotic resistance genes. The inhibitory effect is associated to specific bifidobacterial strains and may be related to the production of thermostable metabolites by these strains. PMID- 17122393 TI - Transcriptional and metabolic responses of Bacillus subtilis to the availability of organic acids: transcription regulation is important but not sufficient to account for metabolic adaptation. AB - The soil bacterium Bacillus subtilis can use sugars or organic acids as sources of carbon and energy. These nutrients are metabolized by glycolysis, the pentose phosphate pathway, and the Krebs citric acid cycle. While the response of B. subtilis to the availability of sugars is well understood, much less is known about the changes in metabolism if organic acids feeding into the Krebs cycle are provided. If B. subtilis is supplied with succinate and glutamate in addition to glucose, the cells readjust their metabolism as determined by transcriptome and metabolic flux analyses. The portion of glucose-6-phosphate that feeds into the pentose phosphate pathway is significantly increased in the presence of organic acids. Similarly, important changes were detected at the level of pyruvate and acetyl coenzyme A (acetyl-CoA). In the presence of organic acids, oxaloacetate formation is strongly reduced, whereas the formation of lactate is significantly increased. The alsSD operon required for acetoin formation is strongly induced in the presence of organic acids; however, no acetoin formation was observed. The recently discovered phosphorylation of acetolactate decarboxylase may provide an additional level of control of metabolism. In the presence of organic acids, both types of analyses suggest that acetyl-CoA was catabolized to acetate rather than used for feeding the Krebs cycle. Our results suggest that future work has to concentrate on the posttranslational mechanisms of metabolic regulation. PMID- 17122394 TI - Vertical transmission of diverse microbes in the tropical sponge Corticium sp. AB - Sponges are host to extremely diverse bacterial communities, some of which appear to be spatiotemporally stable, though how these consistent associations are assembled and maintained from one sponge generation to the next is not well understood. Here we report that a diverse group of microbes, including both bacteria and archaea, is consistently present in aggregates within embryos of the tropical sponge Corticium sp. The major taxonomic groups represented in bacterial 16S rRNA sequences amplified from the embryos are similar to those previously described in a variety of marine sponges. Three selected bacterial taxa, representing proteobacteria, actinobacteria, and a clade including recently described sponge-associated bacteria, were tested and found to be present in all adult samples tested over a 3-year period and in the embryos throughout development. Specific probes were used in fluorescence in situ hybridization to localize cells of the three types in the embryos and mesohyl. This study confirms the vertical transmission of multiple, phylogenetically diverse microorganisms in a marine sponge, and our findings lay the foundation for future work on exploring vertical transmission of specific, yet diverse, microbial assemblages in marine sponges. PMID- 17122395 TI - Green tea polyphenols function as prooxidants to activate oxidative-stress responsive transcription factors in yeasts. AB - Epigallocatechin gallate (EGCG) is the most abundant polyphenolic flavonoid in green tea. Catechin and its derivatives, including EGCG, are widely believed to function as antioxidants. Here we demonstrate that both EGCG and green tea extract (GTE) cause oxidative stress-related responses in the budding yeast Saccharomyces cerevisiae and the fission yeast Schizosaccharomyces pombe under weak alkaline conditions in terms of the activation of oxidative-stress responsive transcription factors. GTE as well as EGCG induced the nuclear localization of Yap1 in S. cerevisiae, which was repressed by the addition of catalase but not by the addition of superoxide dismutase. The same phenomena were observed for the nucleocytoplasmic localization of Msn2 in S. cerevisiae and Pap1, a Yap1 homologue, in S. pombe. The formation of intramolecular disulfide bonds has been proposed to be crucial for the H(2)O(2)-induced nuclear localization of Yap1, and we verified the importance of cysteine residues of Yap1 in response to EGCG and GTE. Additionally, we show that EGCG and GTE produce H(2)O(2) in a weak alkaline medium. Finally, we conclude that tea polyphenols are able to act as prooxidants to cause a response to oxidative stress in yeasts under certain conditions. PMID- 17122396 TI - Homolactate fermentation by metabolically engineered Escherichia coli strains. AB - We report the homofermentative production of lactate in Escherichia coli strains containing mutations in the aceEF, pfl, poxB, and pps genes, which encode the pyruvate dehydrogenase complex, pyruvate formate lyase, pyruvate oxidase, and phosphoenolpyruvate synthase, respectively. The process uses a defined medium and two distinct fermentation phases: aerobic growth to an optical density of about 30, followed by nongrowth, anaerobic production. Strain YYC202 (aceEF pfl poxB pps) generated 90 g/liter lactate in 16 h during the anaerobic phase (with a yield of 0.95 g/g and a productivity of 5.6 g/liter . h). Ca(OH)(2) was found to be superior to NaOH for pH control, and interestingly, significant succinate also accumulated (over 7 g/liter) despite the use of N(2) for maintaining anaerobic conditions. Strain ALS961 (YYC202 ppc) prevented succinate accumulation, but growth was very poor. Strain ALS974 (YYC202 frdABCD) reduced succinate formation by 70% to less than 3 g/liter. (13)C nuclear magnetic resonance analysis using uniformly labeled acetate demonstrated that succinate formation by ALS974 was biochemically derived from acetate in the medium. The absence of uniformly labeled succinate, however, demonstrated that glyoxylate did not reenter the tricarboxylic acid cycle via oxaloacetate. By minimizing the residual acetate at the time that the production phase commenced, the process with ALS974 achieved 138 g/liter lactate (1.55 M, 97% of the carbon products), with a yield of 0.99 g/g and a productivity of 6.3 g/liter . h during the anaerobic phase. PMID- 17122397 TI - Comparison of primer sets for use in automated ribosomal intergenic spacer analysis of aquatic bacterial communities: an ecological perspective. AB - Two primer sets for automated ribosomal intergenic spacer analysis (ARISA) were used to assess the bacterial community composition (BCC) in Lake Mendota, Wisconsin, over 3 years. Correspondence analysis revealed differences in community profiles generated by different primer sets, but overall ecological patterns were conserved in each case. ARISA is a powerful tool for evaluating BCC change through space and time, regardless of the specific primer set used. PMID- 17122398 TI - Nutrient amendments in soil DNA stable isotope probing experiments reduce the observed methanotroph diversity. AB - Stable isotope probing (SIP) can be used to analyze the active bacterial populations involved in a process by incorporating 13C-labeled substrate into cellular components such as DNA. Relatively long incubation times are often used with laboratory microcosms in order to incorporate sufficient 13C into the DNA of the target organisms. Addition of nutrients can be used to accelerate the processes. However, unnatural concentrations of nutrients may artificially change bacterial diversity and activity. In this study, methanotroph activity and diversity in soil was examined during the consumption of 13CH4 with three DNA-SIP experiments, using microcosms with natural field soil water conditions, the addition of water, and the addition of mineral salts solution. Methanotroph population diversity was studied by targeting 16S rRNA and pmoA genes. Clone library analyses, denaturing gradient gel electrophoresis fingerprinting, and pmoA microarray hybridization analyses were carried out. Most methanotroph diversity (type I and type II methanotrophs) was observed in non-amended SIP microcosms. Although this treatment probably best reflected the in situ environmental conditions, one major disadvantage of this incubation was that the incorporation of 13CH4 was slow and some cross-feeding of 13C occurred, thereby leading to labeling of nonmethanotroph microorganisms. Conversely, microcosms supplemented with mineral salts medium exhibited rapid consumption of 13CH4, resulting in the labeling of a less diverse population of only type I methanotrophs. DNA-SIP incubations using water-amended microcosms yielded faster incorporation of 13C into active methanotrophs while avoiding the cross-feeding of 13C. PMID- 17122399 TI - Construction of a Vibrio splendidus mutant lacking the metalloprotease gene vsm by use of a novel counterselectable suicide vector. AB - Vibrio splendidus is a dominant culturable Vibrio in seawater, and strains related to this species are also associated with mortality in a variety of marine animals. The determinants encoding the pathogenic properties of these strains are still poorly understood; however, the recent sequencing of the genome of V. splendidus LGP32, an oyster pathogen, provides an opportunity to decipher the basis of the virulence properties by disruption of candidate genes. We developed a novel suicide vector based on the pir-dependent R6K replicative origin, which potentially can be transferred by RP4-based conjugation to any Vibrio strain and which also carries the plasmid F toxin ccdB gene under control of the PBAD promoter. We demonstrated that this genetic system allows efficient counterselection of integrated plasmids in the presence of arabinose in both V. splendidus and Vibrio cholerae and thus permits efficient markerless allelic replacement in these species. We used this technique to construct several mutants of V. splendidus LGP32, including a derivative with a secreted metalloprotease gene, vsm, deleted. We found that this gene is essential for LGP32 extracellular product toxicity when the extracellular products are injected into oysters but is not necessary for virulence of bacteria in the oyster infection model when bacteria are injected. PMID- 17122400 TI - Rhizonin, the first mycotoxin isolated from the zygomycota, is not a fungal metabolite but is produced by bacterial endosymbionts. AB - Rhizonin is a hepatotoxic cyclopeptide isolated from cultures of a fungal Rhizopus microsporus strain that grew on moldy ground nuts in Mozambique. Reinvestigation of this fungal strain by a series of experiments unequivocally revealed that this "first mycotoxin from lower fungi" is actually not produced by the fungus. PCR experiments and phylogenetic studies based on 16S rRNA gene sequences revealed that the fungus is associated with bacteria belonging to the genus Burkholderia. By transmission electron microscopy, the bacteria were localized within the fungal cytosol. Toxin production and the presence of the endosymbionts were correlated by curing the fungus with an antibiotic, yielding a nonproducing, symbiont-free phenotype. The final evidence for a bacterial biogenesis of the toxin was obtained by the successful fermentation of the endosymbiotic bacteria in pure culture and isolation of rhizonin A from the broth. This finding is of particular interest since Rhizopus microsporus and related Rhizopus species are frequently used in food preparations such as tempeh and sufu. PMID- 17122401 TI - Bacterial genes responsible for the biosynthesis of eicosapentaenoic and docosahexaenoic acids and their heterologous expression. PMID- 17122402 TI - Prevalence of ColE1-like plasmids and colicin K production among uropathogenic Escherichia coli strains and quantification of inhibitory activity of colicin K. AB - Colicin K exhibited pronounced inhibitory activity against uropathogenic Escherichia coli (UPEC) strains. Low prevalence of colicin K production and a relatively high prevalence of ColE1-like plasmids were determined among 215 UPEC strains from Slovenia. Sequencing of the colicin K-encoding pColK-K235 revealed a mosaic structure and the presence of the insertion sequence IS2. PMID- 17122403 TI - Characterization of chimeric Bacillus thuringiensis Vip3 toxins. AB - Bacillus thuringiensis vegetative insecticidal proteins (Vip) are potential alternatives for B. thuringiensis endotoxins that are currently utilized in commercial transgenic insect-resistant crops. Screening a large number of B. thuringiensis isolates resulted in the cloning of vip3Ac1. Vip3Ac1 showed high insecticidal activity against the fall armyworm Spodoptera frugiperda and the cotton bollworm Helicoverpa zea but very low activity against the silkworm Bombyx mori. The host specificity of this Vip3 toxin was altered by sequence swapping with a previously identified toxin, Vip3Aa1. While both Vip3Aa1 and Vip3Ac1 showed no detectable toxicity against the European corn borer Ostrinia nubilalis, the chimeric protein Vip3AcAa, consisting of the N-terminal region of Vip3Ac1 and the C-terminal region of Vip3Aa1, became insecticidal to the European corn borer. In addition, the chimeric Vip3AcAa had increased toxicity to the fall armyworm. Furthermore, both Vip3Ac1 and Vip3AcAa are highly insecticidal to a strain of cabbage looper (Trichoplusia ni) that is highly resistant to the B. thuringiensis endotoxin Cry1Ac, thus experimentally showing for the first time the lack of cross-resistance between B. thuringiensis Cry1A proteins and Vip3A toxins. The results in this study demonstrated that vip3Ac1 and its chimeric vip3 genes can be excellent candidates for engineering a new generation of transgenic plants for insect pest control. PMID- 17122404 TI - In silico model as a tool for interpretation of intestinal infection studies. AB - In nutrition research the number of human in vivo experiments is limited because of the many restrictions and the high costs of testing in humans. Up to now predictive computer models aiming to enhance research have been rare or too complex, with many nonmeasurable adjustable parameters. This study aimed to develop a basic physicochemical computer model for a first quantitative interpretation of results obtained from in vivo intestinal experiments with bacteria. This new modeling approach is validated with results obtained from gut infection studies in vivo. The design of the model is described, and its ability to reproduce experimental data is evaluated. The model predictions are compared with new experimental data. The phenomena that take place in the gastrointestinal tract are summarized by model constants for growth, adherence, and release of bacteria. Although the model is far from describing all details and many processes in the intestine are combined, the model calculation results lead to reasonable conclusions and interesting hypotheses. One of these hypotheses concluded from the model outcomes is that Escherichia coli bacteria have a much lower intestinal growth rate in humans than in rats. Extra laboratory validation experiments proved the reliability of this hypothesis predicted by the model. In addition, the known protective effect of dietary calcium and detrimental effect of clindamycin on the growth and adherence of Salmonella bacteria could be quantified. From these results it is clear that the model enhances the interpretation of in vivo gastrointestinal experiments and will facilitate research trajectories towards new functional foods that improve resistance to pathogenic bacteria in humans. PMID- 17122405 TI - Diversity of Archaea in marine sediments from Skan Bay, Alaska, including cultivated methanogens, and description of Methanogenium boonei sp. nov. AB - Methanogenesis in cold marine sediments is a globally important process leading to methane hydrate deposits, cold seeps, physical instability of sediment, and atmospheric methane emissions. We employed a multidisciplinary approach that combined culture-dependent and -independent analyses with geochemical measurements in the sediments of Skan Bay, Alaska (53 degrees N, 167 degrees W), to investigate methanogenesis there. Cultivation-independent analyses of the archaeal community revealed that uncultivated microbes of the kingdoms Euryarchaeota and Crenarchaeota are present at Skan Bay and that methanogens constituted a small proportion of the archaeal community. Methanogens were cultivated from depths of 0 to 60 cm in the sediments, and several strains related to the orders Methanomicrobiales and Methanosarcinales were isolated. Isolates were psychrotolerant marine-adapted strains and included an aceticlastic methanogen, strain AK-6, as well as three strains of CO(2)-reducing methanogens: AK-3, AK7, and AK-8. The phylogenetic positions and physiological characteristics of these strains are described. We propose a new species, Methanogenium boonei, with strain AK-7 as the type strain. PMID- 17122406 TI - The drought environment: physical, biological and agricultural perspectives. AB - 'Drought' has many meanings in relation to crop production. These range from: statistical (say, the lowest decile of annual rainfall) to a meteorologist; through yield being limited by too little water to an agronomist; to sudden severe water deficits to many molecular biologists. To a farmer, the corresponding management issues, respectively, are risk management (how best to manage a meteorologically drought-prone farm over several years), how best to match cultivar and agronomic operations to the developing growing season, and how best to minimize possible major damage to (say) floral fertility induced by severe water deficits during flowering. All these definitions and the issues they imply are relevant to improving crop production when water is limiting. How can scientists best help? The answers depend on the scales (temporal and spatial) being addressed. Agronomists and breeders, interacting, can help improve components of seasonal water balance in the field, for example, minimizing evaporative losses from the soil surface by better matching the development of a crop to its environment. Physiologists, biochemists, and molecular biologists can help by identifying ways of improving the competence of particular organs. A promising target is floral infertility resulting from water deficits, which results from lesions in tissue, and cellular and molecular processes. Choosing problems whose solutions will have implications in the field and be attractive to farmers requires knowledge of what is important in the field. PMID- 17122407 TI - Genetic and biochemical studies in yeast reveal that the cotton fibre-specific GhCER6 gene functions in fatty acid elongation. AB - 3-ketoacyl-CoA synthase catalyses the initial condensation reaction during fatty acid elongation using malonyl-CoA and long-chain acyl-CoA as substrates. Previously, it was reported that several genes encoding putative cotton 3 ketoacyl-CoA synthases were significantly up-regulated during early cotton fibre development. In this study, GhCER6 cDNA that contains an open reading frame of 1479 bp, encoding a protein of 492 amino acid residues homologous to the Arabidopsis condensing enzyme CER6, was isolated and cloned. In situ hybridization results demonstrated that GhCER6 mRNA was detected only in the elongating wild-type cotton fibre cells. When GhCER6 was transformed to the Saccharomyces cerevisiae elo3 deletion mutation strain that was deficient in the production of 26-carbon fatty acids and displayed a very slow-growth phenotype, the mutant cells were found to divide similarly compared with those of the wild type cells. Further, heterologous expression of GhCER6 restored the viability of the S. cerevisiae haploid elo2 and elo3 double-deletion strain. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis showed that GhCER6 was enzymatically active since the yeast elo2 and elo3 double-deletion mutant expressing the cotton gene produced very-long-chain fatty acids that are essential for cell growth. The results suggest that GhCER6 encodes a functional 3 ketoacyl-CoA synthase. PMID- 17122408 TI - Water permeability differs between growing and non-growing barley leaf tissues. AB - A pressure probe technique and an osmotic swelling assay were used to compare water transport properties between growing and non-growing tissues of leaf three of barley. The epidermis was analysed in planta by pressure probe, whereas (predominantly) mesophyll protoplasts were analysed by osmotic swelling. Hydraulic conductivity (Lp) and, by implication, water permeability (Pf) of epidermal cells was 31% higher in the leaf elongation zone (Lp=0.5+/-0.2 microm s 1 MPa-1; Pf=65+/-25 microm s-1; means+/-SD of n=17 cells) than in the, non growing, emerged leaf zone (Lp=0.4+/-0.1 microm s-1 MPa-1; Pf=50+/-15 microm s-1; n=24; P<0.05). Similarly, water permeability of mesophyll protoplasts was by 55% higher in the elongation compared with emerged leaf zone (Pf=13+/-1 microm s-1 compared with 8+/-1 microm s-1; n=57 and 36 protoplasts, respectively; P<0.01). Within the leaf elongation zone, a small population of larger-sized protoplasts could be distinguished. These protoplasts, which originated most likely from parenchymateous bundle sheath or midrib parenchyma cells, had a three-fold higher water permeability (P<0.001) as mesophyll protoplasts. The effect on Lp and Pf of known aquaporin inhibitors was tested with the pressure probe (Au+, Ag+, Hg2+, phloretin) and the osmotic swelling assay (phloretin). Only phloretin, when applied to protoplasts in the swelling assay caused an average decrease in Pf, but the effect varied between isolations. Technical approaches and cell-type and growth-specific differences in water transport properties are discussed. PMID- 17122409 TI - Cloning and characterization of Arabidopsis and Brassica juncea flavin-containing amine oxidases. AB - Polyamines (PAs) are low molecular weight metabolites involved in various physiological and developmental processes in eukaryotic and prokaryotic cells. The cellular PA level is regulated in part by the action of amine oxidases (AOs) including copper diamine oxidases (DAOs) and flavoprotein polyamine oxidases (PAOs). In this study, the isolation and characterization of flavin amine oxidases (FAOs) from Brassica juncea (BJFAO) and Arabidopsis (ATFAO1) are reported that were clustered in the same group as polyamine oxidases from maize (MPAO) and barley (BPAO1) and monoamine oxidases from mammalian species. ATFAO1 was temporally and spatially regulated in Arabidopsis and showed distinct expression patterns in response to different stress treatments. To investigate the in vivo function of FAO, transgenic Arabidopsis plants expressing sense, antisense, and double-stranded BJFAO RNAs were generated and those with altered activity of FAOs were selected for further characterization. It was found that the shoot regeneration response in transgenic plants was significantly affected by the modulated PA levels corresponding to FAO activities. Tissues that originated from transgenic plants with down-regulated FAO activity were highly regenerative, while those from transgenic plants with upregulated FAO activity were poorly regenerative. The shoot regeneration capacity in these transgenic plants was related to the levels of individual PAs, suggesting that FAO affects shoot regeneration by regulating cellular PAs. Furthermore, it was found that the effect of FAO activity on shoot regeneration was exerted downstream of the Enhancer of Shoot Regeneration (ESR1) gene, which may function in a branch of the cytokinin signalling pathway. PMID- 17122410 TI - Feeling and time: the phenomenology of mood disorders, depressive realism, and existential psychotherapy. AB - Phenomenological research suggests that pure manic and depressive states are less common than mixtures of the two and that the two poles of mood are characterized by opposite ways of experiencing time. In mania, the subjective experience of time is sped up and in depression it is slowed down, perhaps reflecting differences in circadian pathophysiology. The two classic mood states are also quite different in their effect on subjective awareness: manic patients lack insight into their excitation, while depressed patients are quite insightful into their unhappiness. Consequently, insight plays a major role in overdiagnosis of unipolar depression and misdiagnosis of bipolar disorder. The phenomenology of depression also is relevant to types of psychotherapies used to treat it. The depressive realism (DR) model, in contrast to the cognitive distortion model, appears to better apply to many persons with mild to moderate depressive syndromes. I suggest that existential psychotherapy is the necessary corollary of the DR model in those cases. Further, some depressive morbidities may in fact prove, after phenomenological study, to involve other mental states instead of depression. The chronic sub-syndromal depression that is often the long-term consequence of treated bipolar disorder may in fact represent existential despair, rather than depression proper, again suggesting intervention with existential psychotherapeutic methods. PMID- 17122411 TI - PI3K, RSK, and mTOR signal networks for the GST gene regulation. AB - The induction of glutathione S-transferases (GST) represents not only cell detoxification and survival but also cancer prevention. In response to various extracellular stimuli, expression of the gene has been shown to be regulated coordinately by activating the transcription factors in a transcriptional or posttranscriptional manner. Cytoprotective agents induce GST and concomitantly activate the PI3K-Akt/ERK-RSK1-mTOR pathways that activate the transcription factors favoring cell viability. The mechanistic basis and cell signaling for the induction of GST induction by prooxidants and toxicants may be different from that by cytoprotective agents. This paper summarizes the molecular mechanisms of the transcriptional induction of the GST gene orchestrated by a series of transcription factors that recruit coactivators or corepressors. PMID- 17122412 TI - Role of the Kupffer cell in mediating hepatic toxicity and carcinogenesis. AB - Kupffer cells are resident macrophages of the liver and play an important role in its normal physiology and homeostasis as well as participating in the acute and chronic responses of the liver to toxic compounds. Activation of Kupffer cells directly or indirectly by toxic agents results in the release of an array of inflammatory mediators, growth factors, and reactive oxygen species. This activation appears to modulate acute hepatocyte injury as well as chronic liver responses including hepatic cancer. Understanding the role Kupffer cells play in these diverse responses is key to understanding mechanisms of liver injury. Idiosyncratic drug-induced liver disease results in morbidity and mortality, impacting severely on the development of new pharmacological agents. Modulation of the response of Kupffer cells by drugs has been suggested as a cause for the idiosyncratic response. Similarly, liver damage seen in chronic ethanol consumption appears to be modulated by Kupffer cell activation. More recent evidence has noted a contributory role of Kupffer cell activation in the process of hepatic carcinogenesis. Several nongenotoxic carcinogens, for example, activate Kupffer cells resulting in the release of cytokines and/or reactive oxygen species that induce hepatocyte cell proliferation and may enhance clonal expansion of preneoplastic cells leading to neoplasia. Kupffer cells therefore appear to play a central role in the hepatic response to toxic and carcinogenic agents. Taken together, the data presented in this symposium illustrate to the toxicologist the central role played by Kupffer cells in mediating hepatotoxicity. PMID- 17122413 TI - Mitochondrial superoxide production in skeletal muscle fibers of the rat and decreased fiber excitability. AB - Mammalian skeletal muscles generate marked amounts of superoxide (O(2)(.-)) at 37 degrees C, but it is not well understood which is the main source of O(2)(.-) production in the muscle fibers and how this interferes with muscle function. To answer these questions, O(2)(.-) production and twitch force responses were measured at 37 degrees C in mechanically skinned muscle fibers of rat extensor digitorum longus (EDL) muscle. In mechanically skinned fibers, the sarcolemma is removed avoiding potential sources of O(2)(.-) production that are not intrinsically part of the muscle fibers, such as nerve terminals, blood cells, capillaries and other blood vessels in the whole muscle. O(2)(.-) production was also measured in split single EDL muscle fibers, where part of the sarcolemma remained attached, and small bundles of intact isolated EDL muscle fibers at rest, in the presence and absence of modifiers of mitochondrial function. The results lead to the conclusion that mitochondrial production of O(2)(.-) accounts for most of the O(2)(.-) measured intracellularly or extracellularly in skeletal muscle fibers at rest and at 37 degrees C. Muscle fiber excitability at 37 degrees C was greatly improved in the presence of a membrane permeant O(2)(.-) dismutase mimetic (Tempol), demonstrating a direct link between O(2)(.-) production in the mitochondria and muscle fiber performance. This implicates mitochondrial O(2)(.-) production in the down-regulation of skeletal muscle function, thus providing a feedback pathway for communication between mitochondria and plasma membranes that is not directly related to the main function of mitochondria as the power plant of the mammalian muscle cell. PMID- 17122414 TI - Cleavage of the angiotensin II type 1 receptor and nuclear accumulation of the cytoplasmic carboxy-terminal fragment. AB - Our published studies show that the distribution of the ANG II type 1 (AT(1)) receptor (AT(1)R), expressed as a enhanced yellow fluorescent fusion (YFP) protein (AT(1)R/EYFP), is altered upon cellular treatment with ANG II or coexpression with intracellular ANG II. AT(1)R accumulates in nuclei of cells only in the presence of ANG II. Several transmembrane receptors are known to accumulate in nuclei, some as holoreceptors and others as cleaved receptor products. The present study was designed to determine whether the AT(1)R is cleaved before nuclear transport. A plasmid encoding a rat AT(1)R labeled at the amino terminus with enhanced cyan fluorescent protein (CFP) and at the carboxy terminus with EYFP was employed. Image analyses of this protein in COS-7 cells, CCF-STTG1 glial cells, and A10 vascular smooth muscle cells show the two fluorescent moieties to be largely spatially colocalized in untreated cells. ANG II treatment, however, leads to a separation of the fluorescent moieties with yellow fluorescence accumulating in more than 30% of cellular nuclei. Immunoblot analyses of extracts and conditioned media from transfected cells indicate that the CFP domain fused to the extracellular amino-terminal AT(1)R domain is cleaved from the membrane and that the YFP domain, together with the intracellular cytoplasmic carboxy terminus of the AT(1)R, is also cleaved from the membrane bound receptor. The carboxy terminus of the AT(1)R is essential for cleavage; cleavage does not occur in protein deleted with respect to this region. Overexpressed native AT(1)R (nonfusion) is also cleaved; the intracellular 6-kDa cytoplasmic domain product accumulates to a significantly higher level with ANG II treatment. PMID- 17122415 TI - Production of a uniform cellular injury by raster scanning of cells for the study of laser bioeffects. AB - Efforts to understand laser bioeffects in cells and tissues have been hindered by a nonuniform cellular response of the specimen, resulting in graded biochemical effects. In addition, the small beam diameters of commonly used lasers limit the number of cells expressing a response to numbers inadequate for the study of biochemical effects. For a limited emission power, expansion of the beam diameter reduces the irradiance, thus requiring longer exposure durations to produce a cellular response. Cultured human retinal epithelial cells were exposed as a single spot ("tophat" exposure) from a carbon dioxide (CO(2)) laser operating at 10.6 microm or scanned with a raster system and compared with thermal injury produced with heated saline for short periods (1-9 s) at relatively high temperature (55-70 degrees C). Cell viability and induction of the 70 kDa heat shock protein were evaluated as indicators of the cellular response. Initial attempts to use a tophat (uniform energy distribution) exposure resulted in a nonuniform cellular response (and nonuniform energy distribution) due to diffraction effects from the 2-mm selection aperture. However, raster scanning for appropriate times with the CO(2) laser yielded uniform cell viability and heat shock protein synthesis that were comparable to dipping cells in heated saline. Because scanning results in a homogeneous exposure of cells, the described scanning technique may be applied to studies of cellular responses to other lasers to evaluate photochemical and photomechanical effects. PMID- 17122416 TI - Actin disruption inhibits endosomal traffic of P-glycoprotein-EGFP and resistance to daunorubicin accumulation. AB - Intracellular traffic of human P-glycoprotein (P-gp), a membrane transporter responsible for multidrug resistance in cancer chemotherapy, was investigated using a P-gp and enhanced green fluorescent fusion protein (P-gp-EGFP) in human breast cancer MCF-7 cells. The stably expressed P-gp-EGFP from a clonal cell population was functional as a drug efflux pump, as demonstrated by the inhibition of daunorubicin accumulation and the conferring of resistance of the cells to colchicine and daunorubicin. Colocalization experiments demonstrated that a small fraction of the total P-gp-EGFP expressed was localized intracellularly and was present in early endosome and lysosome compartments. P-gp EGFP traffic was shown to occur via early endosome transport to the plasma membrane. Subsequent movement of P-gp-EGFP away from the plasma membrane occurred by endocytosis to the early endosome and lysosome. The component of the cytoskeleton responsible for P-gp-EGFP traffic was demonstrated to be actin rather than microtubules. In functional studies it was shown that in parallel with the interruption of the traffic of P-gp-EGFP, cellular accumulation of the P gp substrate daunorubicin was increased after cells were treated with actin inhibitors, and cell proliferation was inhibited to a greater extent than in the presence of daunorubicin alone. The actin dependence of P-gp traffic and the parallel changes in cytotoxic drug accumulation demonstrated in this study delineates the pathways of P-gp traffic and may provide a new approach to overcoming multidrug resistance in cancer chemotherapy. PMID- 17122417 TI - MRI monitoring of neuroinflammation in mouse focal ischemia. AB - BACKGROUND AND PURPOSE: A growing body of evidence suggests that inflammatory processes are involved in the pathophysiology of stroke. Phagocyte cells, involving resident microglia and infiltrating macrophages, secrete both protective and toxic molecules and thus represent a potential therapeutic target. The aim of the present study was to monitor phagocytic activity after focal cerebral ischemia in mice. METHODS: Ultrasmall superparamagnetic particles of iron oxide (USPIO) were intravenously injected after permanent middle cerebral artery occlusion and monitored by high resolution MRI for 72 hours. RESULTS: We here present the first MRI data showing in vivo phagocyte-labeling obtained in mice with focal cerebral ischemia. USPIO-enhanced MRI kinetic analysis disclosed an inflammatory response surrounding the ischemic lesion and in the contralateral hemisphere via the corpus callosum. The imaging data collected during the first 36 hours postinjury suggested a spread of USPIO-related signal from ipsi- to contralateral hemisphere. Imaging data correlated with histochemical analysis showing inflammation remote from the lesion and ingestion of nanoparticles by microglia/macrophages. CONCLUSIONS: The present study shows that MR-tracking of phagocyte cells is feasible in mice, which may have critical therapeutic implications given the potential neurotoxicity of activated microglia/macrophages in central nervous system disorders. PMID- 17122418 TI - Postischemic augmentation of conducted dilation in cerebral arterioles. AB - BACKGROUND AND PURPOSE: Conducted vasomotor responses likely play an important role in cerebrovascular regulation, but it is unclear how these responses may be affected by ischemia. The purpose of this study was to evaluate the hypothesis that cerebral ischemia and reperfusion (I/R) alters vascular conduction in cerebral arterioles. METHODS: Middle cerebral artery occlusion (MCAO) was induced by an intraluminal filament technique in 4 groups of rats: (A) 2-hour MCAO/24 hour reperfusion (n=14); (B) 2-hour MCAO/1-hour reperfusion (n=7); (C) 1-hour MCAO/24-hour reperfusion (n=6); and (D) 1-hour MCAO/1-hour reperfusion (n=5). Neurological status and infarction (2,3,5-triphenyltetrazolium chloride staining) were evaluated after I/R. Conducted vasomotor responses were assessed in intracerebral branches of the MCA, by following the longitudinal spread of vasodilation or vasoconstriction to localized microapplication of ATP or adenosine. RESULTS: Local microapplication of ATP evoked a biphasic constriction (17+/-3%) and dilation (7+/-2%) response, whereas adenosine elicited only dilation (11+/-2%). These local responses spread longitudinally along sham control arterioles (1 mm conduction distance) with rapid spatial decay. Ischemia followed by 24-hour reperfusion (groups A and C) led to a marked potentiation of conducted dilation responses: dilation to ATP conducted with virtually no decay in I/R arterioles. Augmentation of conductivity was not observed in the 1-hour reperfusion groups (B and D). Moreover, I/R did not alter conducted constriction. CONCLUSIONS: Ischemia-reperfusion led to a specific augmentation of conducted vasodilation in cerebral arterioles. Presumably, enhanced conductivity may improve cerebral perfusion after ischemia. PMID- 17122419 TI - Long-lasting regeneration after ischemia in the cerebral cortex. AB - BACKGROUND AND PURPOSE: Because fibroblast growth factor 2 is a mitogen for central nervous system stem cells, we explored whether long-term fibroblast growth factor 2 delivery to the brain can improve functional outcome and induce cortical neurogenesis after ischemia. METHODS: Rats underwent permanent distal middle cerebral artery occlusion resulting in an ischemic injury limited to the cortex. We used an adeno-associated virus transfection system to induce long-term fibroblast growth factor 2 expression and monitored behavioral and histological changes. RESULTS: Treatment increased the number of proliferating cells and improved motor behavior. Neurogenesis continued throughout 90 days after the ischemia, and the occurrence of newly generated cells with characteristics of neural precursors and immature neurons was most evident 90 days after treatment. CONCLUSIONS: Focal cortical ischemia elicits an ongoing neurogenic response that can be enhanced with fibroblast growth factor 2 leading to improved functional outcome. PMID- 17122420 TI - Macrophage-derived matrix metalloproteinase-2 and -9 promote the progression of cerebral aneurysms in rats. AB - BACKGROUND AND PURPOSE: Mechanisms of initiation, progression and rupture of cerebral aneurysms have not yet been fully understood despite its clinical significance. Matrix metalloproteinases (MMPs) are a family of proteinases which are involved in the remodeling of vascular walls. In the present study, we investigated the significance of MMPs in the progression of cerebral aneurysms. METHODS: Cerebral aneurysms were experimentally induced in 7-week-old male Sprague-Dawley rats. MMP-2 and MMP-9 expression was examined by immunohistochemistry and RT-PCR. Gelatinase activity in aneurysmal walls was assessed by in situ zymography. A selective inhibitor for MMP-2, -9 and -12, tolylsam, was used to examine the effect of inhibition of MMP-2 and MMP-9. RESULTS: Macrophages infiltrated in arterial walls of experimentally induced rat cerebral aneurysms and expressed MMP-2 and -9. Macrophage infiltration and MMP expression was increased with the progression of aneurysms. Gelatinase activity attributable to MMP-2 and MMP-9 increased in arterial walls of rat cerebral aneurysms. Furthermore, tolylsam reduced the ratio of advanced aneurysms in our rat model. CONCLUSIONS: These data suggest that macrophage-derived MMP-2 and -9 may play an important role in the progression of cerebral aneurysms. The findings of this study will shed a new light into the pathogenesis of cerebral aneurysms and highlight the importance of inflammatory response causing the degeneration of extracellular matrix in the process of this disease. PMID- 17122421 TI - Endovascular treatment of experimental aneurysms by use of fibroblast-coated platinum coils: an angiographic and histopathologic study. AB - BACKGROUND AND PURPOSE: The purpose of this study was to determine whether implanting exogenous fibroblasts on platinum coils could enhance intra-aneurysmal fibrosis. Hypotheses included: (1) fibroblast-coated (FBC) platinum coils can improve angiographic results after embolization; and (2) FBC platinum coils can accelerate histological healing of embolized aneurysms. METHODS: Experimental aneurysms in rabbits were embolized with control platinum coils (n=18) or FBC coils (n=18). Subjects were euthanized at 14 days, 1 month, 3 months and 6 months after implantation. Digital subtraction angiography was used to evaluate stability after embolization. Histological samples were examined with a grading system (range, 0 to 12) based on neck and dome healing. RESULTS: Histology total scores and fibrosis ratio at 14 days were significantly greater in the FBC coil group compared with controls (6.6+/-1.9 versus 2.5+/-1.1, 1.2+/-0.6% versus 0.2+/ 0.3%, respectively; P=0.0090). Cavities embolized with FBC coils showed cellular proliferation and thrombus organization, with an endothelialized membrane bridging the neck. There were no differences between groups in the later timepoints. The FBC coil group showed radiographic stability in 11 (61%) cases, coil compaction in 2 (11%) cases, and progressive occlusion in 5 (28%) cases. No progressive occlusion was seen in controls; 3 (17%) of 18 control cases exhibited coil compaction (P=0.0546). CONCLUSIONS: FBC coils can accelerate early histological healing compared with control coils in the rabbit aneurysm model. PMID- 17122422 TI - Serial diffusion tensor MRI after transient and permanent cerebral ischemia in nonhuman primates. AB - BACKGROUND AND PURPOSE: We measured the temporal evolution of the T2 and diffusion tensor imaging parameters after transient and permanent cerebral middle cerebral artery occlusion (MCAo) in macaques, and compared it to standard histological analysis at the study end point. METHODS: Stroke was created in adult male macaques by occluding a middle cerebral artery branch for 3 hours (transient MCAo, n=4 or permanent occlusion, n=3). Conventional MRI and diffusion tensor imaging scans were performed 0 (acute day), 1, 3, 7, 10, 17, and 30 days after MCAo. Animals were euthanized after the final scan and the brains removed for histological analysis. RESULTS: Apparent diffusion coefficient in the lesion was decreased acutely, fractional anisotropy was elevated, and T2 remained normal. Thereafter, apparent diffusion coefficient increased above normal, fractional anisotropy decreased to below normal, T2 increased to a maximum and then declined. Reperfusion at 3 hours accelerated these MRI changes. Only the fractional anisotropy value was significantly different between transient and permanent groups at 30 days. Final MRI-defined fractional lesion volumes were well correlated with corresponding histological lesion volumes. Permanent MCAO animals showed more severe histological damage than their transient MCAO counterparts, especially myelin damage and axonal swelling. CONCLUSIONS: Overall, the MRI evolution of stroke in macaques was closer to what has been observed in humans than in rodent models. This work supports the use of serial MRI in stroke studies in nonhuman primates. PMID- 17122423 TI - From bench to bedside and back: the value of candidate gene association studies in translational research. PMID- 17122424 TI - Pharmacological Induction of Ischemic Tolerance by Glutamate Transporter-1 (EAAT2) Upregulation. AB - BACKGROUND AND PURPOSE: Astrocytic glutamate transporter protein, GLT-1 (EAAT2), recovers extracellular glutamate and ensures that neurons are protected from excess stimulation. Recently, beta-lactam antibiotics, like ceftriaxone (CTX), were reported to induce the upregulation of GLT-1. Here, we investigated ischemic tolerance induction by CTX in an experimental model of focal cerebral ischemia. METHODS: CTX (200 mg/kg per day, IP) was administered for 5 consecutive days before transient focal ischemia, which was induced by intraluminal thread occlusion of the middle cerebral artery for 90 minutes or permanently. RESULTS: Repeated CTX injections enhanced GLT-1 mRNA and protein expressions after 3 and 5 days of treatment, respectively. CTX-pretreated animals showed a reduction in infarct volume by 58% (reperfusion) and 39% (permanent), compared with the vehicle-pretreated animals at 24 hours postischemia (P<0.01). Lower doses of CTX (20 mg/kg per day and 100 mg/kg per day) reduced infarct volumes to a lesser degree. The injection of GLT-1 inhibitor (dihydrokainate) at 30 minutes before ischemia ameliorated the effect of CTX pretreatment. However, CTX administration at 30 minutes after ischemia produced no significant reduction in infarct volume. CTX reduced the levels of proinflammatory cytokines (tumor necrosis factor-alpha, FasL), matrix metalloproteinase (MMP)-9, and activated caspase-9 (P<0.01). In addition, CTX-pretreated animals showed better functional recovery at day 1 to week 5 after ischemia (P<0.05). CONCLUSIONS: This study presents evidence that CTX induces ischemic tolerance in focal cerebral ischemia and that this is mediated by GLT-1 upregulation. PMID- 17122425 TI - Promoter polymorphisms in the plasma glutathione peroxidase (GPx-3) gene: a novel risk factor for arterial ischemic stroke among young adults and children. AB - BACKGROUND AND PURPOSE: Plasma glutathione peroxidase (GPx-3)-deficiency increases extracellular oxidant stress, decreases bioavailable nitric oxide, and promotes platelet activation. The aim of this study is to identify polymorphisms in the GPx-3 gene, examine their relationship to arterial ischemic stroke (AIS) in a large series of children and young adults, and determine their functional molecular consequences. METHODS: We studied the GPx-3 gene promoter from 123 young adults with idiopathic AIS and 123 age- and gender-matched controls by single-stranded conformational polymorphism and sequencing analysis. A second, independent population with childhood stroke was used for a replication study. We identified 8 novel, strongly linked polymorphisms in the GPx-3 gene promoter that formed 2 main haplotypes (H1 and H2). The transcriptional activity of the 2 most prevalent haplotypes was studied with luciferase reporter gene constructs. RESULTS: The H2 haplotype was over-represented in both patient populations and associated with an independent increase in the risk of AIS in young adults (odds ratio=2.07, 95% CI=1.03 to 4.47; P=0.034) and children (odds ratio=2.13, 95% CI=1.23 to 4.90; P=0.027). In adults simultaneously exposed to vascular risk factors, the risk of AIS approximately doubled (odds ratio=5.18, 95% CI=1.82 to 15.03; P<0.001). Transcriptional activity of the H2 haplotype was lower than that of the H1 haplotype, especially after upregulation by hypoxia (normalized relative luminescence: 3.54+/-0.32 versus 2.47+/-0.26; P=0.0083). CONCLUSIONS: These findings indicate that a novel GPx-3 promoter haplotype is an independent risk factor for AIS in children and young adults. This haplotype reduces the gene's transcriptional activity, thereby compromising gene expression and plasma antioxidant and antithrombotic activities. PMID- 17122426 TI - Risk of rebleeding after treatment of acute hydrocephalus in patients with aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Cerebrospinal fluid drainage is often indicated in patients with acute hydrocephalus after aneurysmal subarachnoid hemorrhage but is believed to increase the risk of rebleeding. We studied the risk of rebleeding in patients with subarachnoid hemorrhage during treatment for acute hydrocephalus. METHODS: We included patients with hydrocephalus treated with external ventricular drainage or lumbar punctures within 4 days after the hemorrhage and before aneurysm occlusion. Each treated patient was matched with a control patient with untreated hydrocephalus and a control patient without ventricular enlargement. Patients and controls were matched for interval since subarachnoid hemorrhage, duration of exposure, use of tranexamic acid, clinical condition on admission, and age. We used Cox regression to calculate hazard ratios and we adjusted for rebleeding that had occurred before starting the cerebrospinal fluid drainage. RESULTS: In the group treated with external ventricular drainage, rebleeding occurred in seven of 34 patients (21%) with treatment, in seven of 34 controls (21%) with untreated hydrocephalus, and in six of 34 controls (18%) without hydrocephalus. In the group treated with one or more lumbar punctures, rebleeding occurred in one of 21 patients (5%) with treatment, in three of 21 controls (14%) with untreated hydrocephalus, and in none of the 21 controls without hydrocephalus. The hazard ratios for rebleeding were 1.0 (95% CI: 0.4 to 2.7) for external ventricular drainage treatment and 0.7 (95% CI: 0.1 to 6.4) for lumbar puncture treatment. CONCLUSIONS: This study does not confirm an importantly increased risk of rebleeding during external ventricular drainage or lumbar punctures for acute hydrocephalus after aneurysmal subarachnoid hemorrhage. PMID- 17122427 TI - Intracerebral hemorrhage with severe ventricular involvement: lumbar drainage for communicating hydrocephalus. AB - BACKGROUND AND PURPOSE: The objective was to analyze the feasibility of a lumbar drainage (LD) for a communicating malresorptive hydrocephalus in patients with supratentorial hemorrhage (intracerebral hemorrhage) accompanied by severe ventricular involvement (intraventricular hemorrhage) who required an external ventricular drain (EVD). METHODS: In this retrospective study, 16 patients received an EVD and concurrent LD and were compared with 39 historical patients treated with EVD alone. The duration of required EVD and need for permanent ventriculoperitoneal-shunt were analyzed. RESULTS: LD was inserted after 12 (4 to 18) days. In LD-treated patients, the LD was capable to replace repeated EVD exchanges, resulting in a shorter EVD-duration (12 versus 16 days) compared with patients treated with EVD alone. The overall duration of extracorporal cerebrospinal fluid drainage was longer (16 days EVD versus 21 days EVD+LD) and the frequency of ventriculoperitoneal-shunt lower (18.75% versus 33%; P<0.03) in LD-treated patients. CONCLUSIONS: Our data suggest that LD is safe and feasible for treatment of nonpersistent communicating hydrocephalus after intracerebral hemorrhage. After adequate treatment of the occlusive hydrocephalus using an EVD in the acute phase, LD discloses an alternative for further extracorporal cerebrospinal fluid drainage. PMID- 17122428 TI - The real estate factor: quantifying the impact of infarct location on stroke severity. AB - BACKGROUND AND PURPOSE: The severity of the neurological deficit after ischemic stroke is moderately correlated with infarct volume. In the current study, we sought to quantify the impact of location on neurological deficit severity and to delineate this impact from that of volume. METHODS: We developed atlases consisting of location-weighted values indicating the relative importance in terms of neurological deficit severity for every voxel of the brain. These atlases were applied to 80 first-ever ischemic stroke patients to produce estimates of clinical deficit severity. Each patient had an MRI and National Institutes of Health Stroke Scale (NIHSS) examination just before or soon after hospital discharge. The correlation between the location-based deficit predictions and measured neurological deficit (NIHSS) scores were compared with the correlation obtained using volume alone to predict the neurological deficit. RESULTS: Volume-based estimates of neurological deficit severity were only moderately correlated with measured NIHSS scores (r=0.62). The combination of volume and location resulted in a significantly better correlation with clinical deficit severity (r=0.79, P=0.032). CONCLUSIONS: The atlas methodology is a feasible way of integrating infarct size and location to predict stroke severity. It can estimate stroke severity better than volume alone. PMID- 17122429 TI - Chronic treatment with minocycline preserves adult new neurons and reduces functional impairment after focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: Evidence suggests that activated microglia are detrimental to the survival of new hippocampal neurons, whereas blocking inflammation has been shown to restore hippocampal neurogenesis after cranial irradiation and seizure. The aim of this current study is to determine the effect of minocycline on neurogenesis and functional recovery after cerebral focal ischemia. METHODS: Four days after temporary middle cerebral artery occlusion, minocycline was administered intraperitoneally for 4 weeks. BrdU was given on days 4 to 7 after middle cerebral artery occlusion to track cell proliferation. The number of remaining new neurons and activated microglia were quantified in the dentate gyrus. Infarct volume was measured to assess the treatment effect of minocycline. Motor and cognitive functions were evaluated 6 weeks after middle cerebral artery occlusion. RESULTS: Minocycline delivered 4 days after middle cerebral artery occlusion for 4 weeks did not result in reduction in infarct size but significantly decreased the number of activated microglia in the dentate gyrus. Minocycline also significantly increased the number of newborn neurons that coexpressing BrdU and NeuN without significantly affecting progenitor cell proliferation in the dentate gyrus. Lastly, minocycline significantly improved motor coordination on the rotor rod, reduced the preferential use of the unaffected limb during exploration, reduced the frequency of footfalls in the affected limb when traversing on a horizontal ladder, and improved spatial learning and memory in the water maze test. CONCLUSIONS: Minocycline reduces functional impairment caused by cerebral focal ischemia. The improved function is associated with enhanced neurogenesis and reduced microglia activation in the dentate gyrus and possibly improved neural environment after chronic treatment with minocycline. PMID- 17122430 TI - Cost-effectiveness of intravenous thrombolysis with alteplase within a 3-hour window after acute ischemic stroke. AB - BACKGROUND AND PURPOSE: The aim of this study was to assess the costs and cost effectiveness of intravenous thrombolysis treatment with alteplase (Actilyse) of acute ischemic stroke with 24-hour in-house neurology coverage and use of magnetic resonance imaging. METHODS: A health economic model was designed to calculate the marginal cost-effectiveness ratios for time spans of 1, 2, 3 and 30 years. Effect data were extracted from a meta-analysis of six large-scale randomized and placebo-controlled studies of thrombolytic therapy with alteplase. Cost data were extracted from thrombolysis treatment at Aarhus Hospital, Denmark, and from previously published literature. RESULTS: The calculated cost effectiveness ratio after the first year was $55,591 US per quality-adjusted life year (base case). After the second year, computation of the cost-effectiveness ratio showed that thrombolysis was cost-effective. The long-term computations (30 years) showed that thrombolysis was a dominant strategy compared with conservative treatment given the model premises. CONCLUSIONS: A high-quality thrombolysis treatment with 24-hour in-house neurology coverage and magnetic resonance imaging might not be cost-effective in the short term compared with conservative treatment. In the long term, there are potentially large-scale health economic cost savings. PMID- 17122431 TI - Cholinergic neuronal deficits in CADASIL. AB - BACKGROUND AND PURPOSE: Previous evidence from MRI and acetylcholinesterase histochemistry suggests cholinergic fibers are affected in cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). METHODS: As a measure of cholinergic function, we assessed choline acetyltransferase (ChAT) activities in the frontal and temporal neocortices and the immunocytochemical distribution of ChAT and p75 neurotrophin receptor (P75(NTR)) by in vitro imaging in the nucleus basalis of Meynert of CADASIL subjects. RESULTS: ChAT activities were significantly reduced by 60% to 70% in frontal and temporal cortices of CADASIL cases, as were ChAT and P75(NTR) immunoreactivities in the nucleus basalis. CONCLUSIONS: Our findings suggest cholinergic neuronal impairment in CADASIL and implicate cholinomimetic therapy for subcortical vascular dementias. PMID- 17122432 TI - Effects of splinting on wrist contracture after stroke: a randomized controlled trial. AB - BACKGROUND AND PURPOSE: Splints are commonly applied to the wrist and hand to prevent and treat contracture after stroke. However, there have been few randomized trials of this intervention. We sought to determine whether wearing a hand splint, which positions the wrist in either a neutral or an extended position, reduces wrist contracture in adults with hemiplegia after stroke. METHODS: Sixty-three adults who had experienced a stroke within the preceding 8 weeks participated. They were randomized to either a control group (routine therapy) or 1 of 2 intervention groups (routine therapy plus splint in either a neutral or an extended wrist position). Splints were worn overnight for, on average, between 9 and 12 hours, for 4 weeks. The primary outcome, measured by a blinded assessor, was extensibility of the wrist and long finger flexor muscles (angle of wrist extension at a standardized torque). RESULTS: Neither splint appreciably increased extensibility of the wrist and long finger flexor muscles. After 4 weeks, the effect of neutral wrist splinting was to increase wrist extensibility by a mean of 1.4 degrees (95% CI, -5.4 degrees to 8.2 degrees), and splinting the wrist in extension reduced wrist extensibility by a mean of 1.3 degrees (95% CI, -4.9 degrees to 2.4 degrees) compared with the control condition. CONCLUSIONS: Splinting the wrist in either the neutral or extended wrist position for 4 weeks did not reduce wrist contracture after stroke. These findings suggest that the practice of routine wrist splinting soon after stroke should be discontinued. PMID- 17122433 TI - Is intra-arterial thrombolysis safe after full-dose intravenous recombinant tissue plasminogen activator for acute ischemic stroke? AB - BACKGROUND AND PURPOSE: The optimal approach for acute ischemic stroke patients who do not respond to intravenous recombinant tissue plasminogen activator (IV rt PA) is uncertain. This study evaluated the safety and response to intra-arterial thrombolytics (IATs) in patients unresponsive to full-dose IV rt-PA. METHODS: A case series from a prospectively collected database on consecutive acute ischemic stroke patients treated with IATs after 0.9 mg/kg IV rt-PA during a 7-year interval was collected. Primary outcome measures included symptomatic intracranial hemorrhage and mortality. As indicators of response, secondary outcome measures were recanalization and discharge disposition. RESULTS: Sixty nine patients (mean+/-SD age, 60+/-13 years; range, 26 to 85 years; 55% male) with a median pretreatment National Institutes of Health Stroke Scale score of 18 (range, 6 to 39) were included. IV rt-PA was started at 124+/-32 minutes (median, 120 minutes) and IAT, at 288+/-57 minutes (median, 285 minutes). IATs consisted of reteplase (n=56), alteplase (n=7), and urokinase (n=6), with an average total dosage of 2.8 U, 8.6 mg, and 700 000 U, respectively. Symptomatic intracranial hemorrhage occurred in 4 of 69 (5.8%) patients; 3 cases were fatal. Recanalization was achieved in 50 (72.5%) and a favorable outcome (home or inpatient rehabilitation) in 38 (55%). CONCLUSIONS: IAT therapy after full-dose IV rt-PA in patients with persisting occlusion and/or lack of clinical improvement appears safe compared with IV rt-PA alone or low-dose IV rt-PA followed by IAT. A high rate of recanalization and favorable outcome can be achieved. PMID- 17122434 TI - Enhanced thrombogenesis but not platelet activation is associated with transcatheter closure of patent foramen ovale in patients with cryptogenic stroke. AB - BACKGROUND AND PURPOSE: No studies have yet determined whether antiplatelet or anticoagulant therapy is the more appropriate treatment after transcatheter closure of patent foramen ovale (PFO) in patients with cryptogenic stroke. The objective of this study was to prospectively evaluate the presence, degree, and timing of activation of the platelet and coagulation systems after transcatheter closure of PFO in patients with cryptogenic stroke. METHODS: Twenty-four consecutive patients (mean age, 44+/-10 years; 11 men) with previous cryptogenic stroke who had undergone successful transcatheter closure of PFO were included in the study. Prothrombin fragment 1+2 (F1+2) and thrombin-antithrombin III (TAT) were used as markers of coagulation activation, and soluble P-selectin and soluble CD40 ligand were used as markers of platelet activation. Measurements of all hemostatic markers were taken at baseline just before the procedure and at 7, 30, and 90 days after device implantation. RESULTS: F1+2 and TAT levels increased from 0.41+/-0.16 nmol/L and 2.34+/-1.81 ng/mL, respectively, at baseline to a maximal value of 0.61+/-0.16 nmol/L and 4.34+/-1.83 ng/mL, respectively, at 7 days, gradually returning to baseline levels at 90 days (P<0.001 for both markers). F1+2 and TAT levels at 7 days after PFO closure were higher than those obtained in a group of 25 healthy controls (P<0.001 for both markers). Levels of soluble P-selectin and soluble CD40 ligand did not change at any time after PFO closure. CONCLUSIONS: Transcatheter closure of PFO is associated with significant activation of the coagulation system, with no increase in platelet activation markers. These findings raise the question of whether optimal antithrombotic treatment after PFO closure should be short-term anticoagulant rather than antiplatelet therapy. PMID- 17122435 TI - Thrombolysis of basilar artery occlusion--intra-arterial or intravenous: is there really no difference? PMID- 17122437 TI - Asymptomatic hemorrhage after thrombolysis may not be benign: prognosis by hemorrhage type in the Canadian alteplase for stroke effectiveness study registry. AB - BACKGROUND AND PURPOSE: There is ongoing controversy about the impact of hemorrhagic transformation after thrombolysis on long-term functional outcome. We sought to study the relation between the type of hemorrhagic transformation on CT scans and functional outcome. METHODS: Data were obtained from the Canadian Alteplase for Stroke Effectiveness Study. This study was established as a registry to prospectively collect data for acute stroke patients receiving intravenous alteplase within 3 hours from stroke onset between February 1999 and June 2001. Follow-up was completed at 90 days, and good functional outcome was defined as a modified Rankin Scale score of 0 or 1. Copies of head CT scans obtained at 24 to 48 hours after starting treatment were read in consensus by a central reading panel consisting of 1 neuroradiologist and 1 stroke neurologist. According to European Cooperative Acute Stroke Study criteria, hemorrhagic transformation was classified as none, hemorrhagic infarction (HI-1 and HI-2), or parenchymal hematoma (PH-1 and PH-2). We compared outcome across groups and performed a multivariable analysis including previously determined important predictors of good outcome in acute ischemic stroke. RESULTS: From 1135 patients enrolled at 60 centers across Canada, 954 follow-up CT scans were assessable. We observed some hemorrhagic transformation in 259 of 954 (27.1%) patients (110 HI 1, 57 HI-2, 48 PH-1, and 44 PH-2). Proportions of patients with good outcome were 41% with no hemorrhagic transformation, 30% with HI-1, 17% with HI-2, 15% with PH 1, and 7% with PH-2 (P<0.0001, chi(2) test). After adjustment for age, baseline serum glucose, baseline Alberta Stroke Program Early CT score, and baseline National Institutes of Health Stroke Scale score, HI-1 was not a predictor of outcome. However, HI-2 (odds ratio=0.38, 95% CI=0.17 to 0.83), PH-1 (odds ratio=0.32, 95% CI=0.12 to 0.80), and PH-2 (odds ratio=0.14, 95% CI=0.04 to 0.48) were all negative predictors of outcome. CONCLUSIONS: The likelihood of a poor outcome after thrombolysis was proportional to the extent of hemorrhage on CT scans. HI grades of hemorrhagic transformation may not be benign. PMID- 17122438 TI - Genome-wide linkage scan of common stroke in families from northern Sweden. AB - BACKGROUND AND PURPOSE: Taking advantage of low genetic variations in northern Sweden, we performed a genome-wide linkage scan to investigate the susceptibility loci for common forms of stroke. METHODS: Fifty-six families, containing multiple cases of stroke and whose data had been previously used to replicate linkage to the phosphodiesterase 4D locus on chromosome 5q, were genotyped in a genome-wide scan. Fine mapping was performed, and subsequently 53 additional families from the same region were genotyped over the candidate regions. RESULTS: Linkage calculations were performed by using 3 different disease models, from a very broad (all stroke cases defined by World Health Organization MONICA criteria) to a narrower (ischemic stroke only) stroke phenotype. With all models, nonparametric multipoint linkage analysis yielded allele-sharing log of the odds (LOD) scores >1.2 at 9 locations: 1p34, 5q13, 7q35, 9q22, 9q34, 13q32, 14q32, 18p11, and 20q13. The highest allele-sharing LOD scores were obtained on chromosomes 5q (previously reported), 1p (LOD=2.09), and 18p (LOD=2.14). Fine mapping resulted in increased allele-sharing LOD scores for chromosome 5q (previously reported) and 9q22 (LOD=1.56), but all others decreased. Combining these initial results with a subsequent analysis of 53 additional families, we obtained the highest allele-sharing LOD scores on chromosomes 5q, 13q, and 18p, although none reached the initial genome-wide allele-sharing LOD scores. CONCLUSIONS: Genetic analysis of stroke in families from northern Sweden did not identify any new major stroke loci. This indicates that multiple minor susceptibility loci in addition to the previously known locus on chromosome 5 could contribute to the disease. PMID- 17122439 TI - Identification and clinical impact of impaired cerebrovascular autoregulation in patients with malignant middle cerebral artery infarction. AB - BACKGROUND AND PURPOSE: To study cerebrovascular autoregulation and its impact on clinical course in patients with impending malignant middle cerebral artery infarction, we used invasive multimodal neuromonitoring, including measurement of cerebral perfusion pressure, tissue oxygen pressure, and microdialysis. METHODS: Fifteen patients with a stroke that involved >50% of the middle cerebral artery territory were included. Probes were placed into the ipsilateral frontal lobe. Autoregulation was assessed by calculation of the cerebral perfusion pressure oxygen reactivity index (COR) and the correlation coefficient (R) of cerebral perfusion pressure and tissue oxygen pressure at 24 and 72 hours after stroke. RESULTS: COR and R at 24 hours after stroke were higher in the 8 patients with a malignant course (ie, massive edema formation) compared with the 7 patients with a benign course (COR, 1.99+/-1.46 versus 0.68+/-0.29; R, 0.49+/-0.28 versus 0.06+/-0.31; P<0.05), indicating impaired autoregulation in the malignant course group. At 72 hours, further increases in COR and R were observed in the malignant course group in contrast to the benign course group with stable values over time (COR, 3.31+/-2.38 versus 0.75+/-0.31; R, 0.75+/-011 versus 0.36+/-0.27; P<0.05). With a COR of 0.99, a cutoff value for prediction of a malignant course was found. The lactate-pyruvate ratio was higher in patients with a malignant compared with a benign course at both time points. COR, R, and the lactate pyruvate ratio showed significant correlations with outcome parameters as a midline shift on cranial computed tomography and score on the modified Rankin scale after 3 months. CONCLUSIONS: We found early impairment of cerebrovascular autoregulation in peri-infarct tissue of patients who developed malignant brain edema, whereas autoregulation was preserved in patients with a benign course. Impaired cerebral autoregulation seems to play a key role for development of a malignant course and might serve as a predictive marker. Impaired cerebral autoregulation also accentuates the need for consequent adjustment of cerebral perfusion pressure in patients with impaired autoregulation. PMID- 17122440 TI - Regulation of smooth muscle cell proliferation by beta-catenin/T-cell factor signaling involves modulation of cyclin D1 and p21 expression. AB - We previously observed that stimulation of vascular smooth muscle cell (VSMC) proliferation with growth factors is associated with dismantling of cadherin junctions and nuclear translocation of beta-catenin. In this study we demonstrate directly that growth factors stimulate beta-catenin/T-cell factor (TCF) signaling in primary VSMCs. To determine whether beta-catenin/TCF signaling regulates VSMC proliferation via modulation of the beta-catenin/TCF responsive cell cycle genes, cyclin D1 and p21, we inhibited beta-catenin/TCF signaling by adenoviral-mediated over-expression of N-Cadherin, ICAT (an endogenous inhibitor of beta-catenin/TCF signaling), or a dominant negative (dn) mutant of TCF-4. N-cadherin, ICAT or dnTCF-4 over-expression significantly reduced proliferation of isolated human VSMCs by approximately 55%, 80%, and 45% respectively. Similar effects were observed in human saphenous vein medial segments where proliferation was reduced by approximately 55%. Transfection of dnTCF-4 in the ISS10 human VSMC line significantly lowered TCF and cyclin D1 reporter activity but significantly elevated p21 reporter activity, indicating regulation of these genes by beta catenin/TCF signaling. In support of this, over-expression of N-cadherin, ICAT or dnTCF-4 in isolated human VSMCs significantly lowered levels of cyclin D1 mRNA and protein levels. In contrast, over-expression of N-Cadherin, ICAT or dnTCF4 significantly elevated p21 mRNA and protein levels. In summary, we have demonstrated that increasing N-cadherin and inhibiting beta-catenin/TCF signaling reduces VSMC proliferation, decreases the expression of cyclin D1 and increases levels of the cell cycle inhibitor, p21. We therefore suggest that the N-cadherin and beta-catenin/TCF signaling pathway is a key modulator of VSMC proliferation via regulation of these 2 beta-catenin/TCF responsive genes. PMID- 17122441 TI - Bone marrow Oct3/4+ cells differentiate into cardiac myocytes via age-dependent paracrine mechanisms. AB - The mechanisms that govern the capacity of the bone marrow stem cells to generate cardiac myocytes are still unknown. Herein we demonstrate that the cardiomyogenic potential of bone marrow-derived Oct3/4(+)/cKit(+/-)/CXCR4(+/-)/CD34(-)/Sca1(-) cells is governed by age-dependent paracrine/juxtacrine platelet-derived growth factor (PDGF) pathways. Specifically, bone marrow cell cultures from both 3- and 18-month-old mice formed aggregates of Oct3/4(+) cells circumscribed by PDGFRalpha(+)/Oct3/4(-)/Sca1(+) cells. In young (3-month) bone marrow cell cultures, induction of PDGF-AB preceded the induction of cardiac genes and was required for the generation of cardiomyogenesis. Indeed, in old (18-month) cultures, diminished PDGF-B induction was associated with impaired cardiomyogenic potential, despite having Oct3/4 levels similar to those in the young cells. Importantly, supplementation with PDGF-AB specifically restored the cardiac differentiation capacity of the old bone marrow cells. Together these results demonstrate that, regardless of age, the bone marrow niche contains Oct3/4 stem cells that are capable of differentiating into cardiac myocytes. Moreover, this differentiation is governed by age-dependent PDGF-AB-mediated paracrine/juxtacrine pathways that may be essential in the translation of bone marrow cell-mediated cardiomyogenesis. PMID- 17122442 TI - C-Peptide in insulin resistance and vascular complications: teaching an old dog new tricks. PMID- 17122443 TI - Inflammation and coagulation in the cardiovascular system: the contribution of influenza. PMID- 17122444 TI - A novel approach to the suppression of atherosclerosis by Fcgamma receptor blockade. PMID- 17122445 TI - Gap junctions: clarifying the complexities of connexins and conduction. PMID- 17122446 TI - Regulation of vascular inflammation and remodeling by ETS factors. AB - The ETS (E26 Transformation-specific Sequence) factors are comprised of a family of transcription factors that share a highly conserved DNA binding domain. Although originally described for their role as protooncogenes in the development of several types of human cancer, they have subsequently been shown to regulate a wide variety of biological processes including cellular growth and differentiation under normal and pathological conditions. As transcription factors, they can either function as activators or repressors of gene expression. Several ETS family members are expressed in cells of vascular origin, including endothelial cells and vascular smooth muscle cells, where they regulate the expression of a number of vascular-specific genes. In the past few years, emerging evidence supports a novel role for selected ETS family members in the regulation of vascular inflammation and remodeling. ETS factor expression can be induced by proinflammatory cytokines, growth factors, and vasoactive peptides. Examples of some of the target genes regulated by ETS factors include adhesion molecules, chemokines, and matrix metalloproteinases. Targeted disruption of selected ETS family members such as Ets-1 in mice is associated with marked reductions in the recruitment of inflammatory cells and vascular remodeling in response to systemic administration of the vasoactive peptide angiotensin II. The purpose of this review is to provide an overview of recent advances that have been made in defining a role for selected members of the ETS transcription factor family in the regulation of vascular-specific gene expression, vascular inflammation, and remodeling. PMID- 17122447 TI - Telomere biology and cardiovascular disease. AB - Accumulation of cellular damage with advancing age leads to atherothrombosis and associated cardiovascular disease. Ageing is also characterized by shortening of the DNA component of telomeres, the specialized genetic segments located at the end of eukaryotic chromosomes that protect them from end-to-end fusions. By inducing genomic instability, replicative senescence and apoptosis, shortening of the telomeric DNA is thought to contribute to organismal ageing. In this Review, we discuss experimental and human studies that have linked telomeres and associated proteins to several factors which influence cardiovascular risk (eg, estrogens, oxidative stress, hypertension, diabetes, and psychological stress), as well as to neovascularization and the pathogenesis of atherosclerosis and heart disease. Two chief questions that remain unanswered are whether telomere shortening is cause or consequence of cardiovascular disease, and whether therapies targeting the telomere may find application in treating these disorders (eg, cell "telomerization" to engineer blood vessels of clinical value for bypass surgery, and to facilitate cell-based myocardial regeneration strategies). Given that most research to date has focused on the role of telomerase, it is also of up most importance to investigate whether alterations in additional telomere associated proteins may contribute to the pathogenesis of cardiovascular disease. PMID- 17122448 TI - A causal role for endothelin-1 in the vascular adaptation to skeletal muscle deconditioning in spinal cord injury. AB - OBJECTIVE: Endothelin-1 (ET-1) contributes to the increased peripheral resistance in heart failure and hypertension. Physical inactivity is associated with cardiovascular disease and characterized by increased vascular tone. In this study, we assess the contribution of ET-1 to the increased vascular tone in the extremely deconditioned legs of spinal cord-injured (SCI) individuals before and after exercise training. METHODS AND RESULTS: In 8 controls and 8 SCI individuals, bilateral thigh blood flow was measured by plethysmography before and during the administration of an ET(A)/ET(B)-receptor blocker into the femoral artery. In SCI, this procedure was repeated after 6 weeks of electro-stimulated training. In a subset of SCI (n=4), selective ET(A)-receptor blockade was performed to determine the role of the ET(A)-receptors. In controls, dual ET receptor blockade increased leg blood flow at the infused side (10%, P<0.05), indicating a small contribution of ET-1 to leg vascular tone. In SCI, baseline blood flow was lower compared with controls (P=0.05). In SCI, dual ET-receptor blockade increased blood flow (41%, P<0.001). This vasodilator response was significantly larger in SCI compared with controls (P<0.001). The response to selective ET(A)-receptor blockade was similar to the effect of dual blockade. Electro-stimulated training normalized baseline blood flow in SCI and reduced the response to dual ET-receptor blockade in the infused leg (29%, P=0.04). CONCLUSIONS: ET-1 mediates the increased vascular tone of extremely inactive legs of SCI individuals by increased activation of ET(A)-receptors. Physical training reverses the ET-1-pathway, which normalizes basal leg vascular tone. PMID- 17122449 TI - Hemodynamic shear stresses in mouse aortas: implications for atherogenesis. AB - OBJECTIVE: The hemodynamic environment is a determinant of susceptibility to atherosclerosis in the vasculature. Although mouse models are commonly used in atherosclerosis studies, little is known about local variations in wall shear stress (WSS) in the mouse and whether the levels of WSS are comparable to those in humans. The objective of this study was to determine WSS values in the mouse aorta and to relate these to expression of gene products associated with atherosclerosis. METHODS AND RESULTS: Using micro-CT and ultrasound methodologies we developed a computational fluid dynamics model of the mouse aorta and found values of WSS to be much larger than those for humans. We also used a quantum dot based approach to study vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 expression on the aortic intima and demonstrated that increased expression for these molecules occurs where WSS was relatively low for the mouse. CONCLUSIONS: Despite large differences in WSS in the two species, the spatial distributions of atherogenic molecules in the mouse aorta are similar to atherosclerotic plaque localization found in human aortas. These results suggest that relative differences in WSS or in the direction of WSS, as opposed to the absolute magnitude, may be relevant determinants of flow-mediated inflammatory responses. PMID- 17122450 TI - Implementation of colorectal cancer screening. PMID- 17122451 TI - Chronic idiopathic thrombocytopenic purpura: present strategy, guidelines and new insights. AB - Idiopathic thrombocytopenic purpura (ITP) is an immune-mediated thrombocytopenia. The diagnosis is made after exclusion of other secondary causes of thrombocytopenic disorders. The primary treatment goal is to prevent severe bleeding rather than achieve normal platelet counts. In adults ITP usually has an insidious onset and chronic course. Although ITP is a relatively common haematological disorder, there are important unresolved issues in its management, especially for chronic refractory ITP patients. New therapeutic agents have changed strategies for ITP treatment. This article reviews the treatment indications and options of chronic ITP in adults in the literature and compares them with the treatment indications and treatment options used by the Dutch internist. PMID- 17122452 TI - The consequences of lost gallstones during laparoscopic cholecystectomy. AB - Laparoscopic cholecystectomy has become the preferred surgical technique for symptomatic gallstone disease. The technique generally is safe. probably one of the most common intra-operative complications is gallbladder perforation with stones spreading into the peritoneal cavity. In this paper the sequelae of lost gallstones after laparoscopic cholecystectomy and the diagnostic problems facing the clinician are reviewed. Abscesses and fistula formation in the abdominal wall occur. A long delay can be present between the initial operation and the complications of the lost stones. Although rupture of the gallbladder is usually noticed during preparation and retrieval, the surgeon may not be aware of losing stones. due to the long delay, the occurrence of intra-abdominal abscesses and fistula is often not linked to the prior procedure. PMID- 17122453 TI - The frequency of a positive family history for colorectal cancer: a population based study in the Netherlands. AB - BACKGROUND: Subjects with a positive family history of colorectal cancer (CRC) have an increased risk of developing CRC themselves. This risk depends on the number of affected relatives and the age at diagnosis. AIM: The aim of this study was to assess the prevalence of a positive family history of CRC, within a random cohort among the dutch population. METHODS: A total of 5072 subjects aged between 45 and 70 years were invited to fill in an anonymous questionnaire about the occurrence of CRC in their first-degree relatives (FDR). RESULTS: The questionnaire was returned by 3973 subjects (78.3%). Thirty responders (0.8%) had CRC themselves. Of all unaffected responders, 441 (11.2%) subjects reported a positive family history of CRC. Ninety (2.3%) responders reported having an FDR with CRC diagnosed before the age of 50, or reported two or more FDRs with CRC. CONCLUSION: The prevalence of a positive family history of CRC is substantial. Identification of this high-risk group by obtaining a thorough family history is the first step in targeting preventive measures. PMID- 17122454 TI - Dutch endoscopic capacity in the era of colorectal cancer screening. AB - BACKGROUND: Future colorectal cancer (CRC) screening programmes should not (greatly) interfere with regular health care. Hence, we analysed the Dutch endoscopic practice to provide a clear insight into endoscopic workload and manpower with a special emphasis on the current ability to facilitate a successful implementation of a faecal occult blood test (FOBT)-based nationwide CRC screening programme. METHODS: A questionnaire was sent to all Dutch endoscopy units (n = 100) in the spring of 2005. The questionnaire included topics ranging from the numbers and specifications of endoscopies performed in 2004 and the numbers of endoscopists per unit to expected vacancies for gastroenterologists and waiting times. RESULTS: The response rate was 98%, representing a total of 49,253 hospital beds. overall, a 26% increase in the number of endoscopies from 325,000 in 1999 to almost 410,000 in 2004 was found, accompanied by a 25% increase in manpower. The total number of endoscopists was 598. regional differences were observed in the number of endoscopists, the total number of endoscopies and colonoscopies, and the number of endoscopies per endoscopist. A biannual FOBT-based screening programme would yield an additional workload of 25,385 colonoscopies a year amounting to a 22% increase in the total number of colonoscopies performed. However, the workload per unit would only have to increase by five extra colonoscopies a week. CONCLUSION: Whereas an FOBT-based CRC screening programme is currently feasible without strongly interfering with regular health care, future plans regarding the scale and preferred mode of screening should incorporate solid data on the (regional) endoscopic capacity and manpower needed for a successful implementation. PMID- 17122455 TI - Vascular type of Ehlers-Danlos syndrome in a patient with ruptured aneurysm of the splenic artery. AB - A 39-year-old woman presented with a ruptured aneurysm of the splenic artery. The postoperative course was complicated by poor wound healing. This, in combination with a history of easy bruising and joint hypermobility, made us consider a connective tissue disease as underlying cause. The vascular type of Ehlers-Danlos syndrome was diagnosed by identifying collagen III deficiency and the corresponding gene mutation in cultured fibroblasts from a skin biopsy. PMID- 17122456 TI - Risk factors of acute hepatic failure during antituberculosis treatment: two cases and literature review. AB - Hepatotoxicity is a well-known side effect of antituberculosis treatment (ATT). If not recognised in time, drug-induced hepatitis can develop, which may rapidly progress to acute liver failure. We describe two patients with acute hepatic failure caused by ATT, whose pretreatment liver function had been normal. Both patients successfully underwent liver transplantation. Possible risk factors predisposing towards ATT-induced hepatic failure were evaluated, and at least four risk factors were present in these patients. Although available guidelines do not advocate routine monitoring of liver function during ATT unless baseline values are elevated or in the case of pre-existent liver disease, this is nevertheless common practice. Liver function should always be measured in patients who develop symptoms during ATT, and rising liver function parameters should prompt immediate action to prevent the occurrence of liver failure. This report underscores that regular monitoring of liver function parameters and adherence to guidelines is especially important in patients with risk factors for ATT-induced liver disease. An evaluation of chronic viral hepatitis in risk groups before starting ATT could be worthwhile. PMID- 17122457 TI - 'Nutrothorax' due to misplacement of a nasogastric feeding tube. AB - we report a serious complication of blind nasogastric feeding tube insertion in a 65-year-old female patient, which was overlooked and caused severe respiratory failure. PMID- 17122458 TI - Status epilepticus and Hashimotos encephalopathy. PMID- 17122459 TI - CT colonography to visualise the whole colon can be complementary to incomplete colonoscopy. PMID- 17122460 TI - The general professional education of the physician. PMID- 17122461 TI - Resident duty hours reform: are we there yet? AB - In 2003, the Accreditation Council for Graduate Medical Education (ACGME) implemented resident duty hours restrictions to address growing concerns about medical errors and resident well-being. Many anticipated that resident duty hours restrictions would improve the quality and safety of care by minimizing the detrimental effects of fatigue on resident performance. Others were concerned that the fundamental clinical and educational principle of continuity of care would be lost or at least eroded, and that more frequent "hand-offs" might result in more clinical errors. Some lamented the loss of the total-emersion residency experience that serves as a forging process to temper the mind and body to create a finely honed clinician. The author draws from the literature to examine the effects of the ACGME resident duty hours restrictions three years after their implementation. From the perspectives of resident perceptions, attending perceptions, organizational approaches, and unintended consequences, the author concludes that far more than simple control of duty hours will be required to achieve the goals of clinical excellence, educational excellence, resident well being, and professionalism. PMID- 17122462 TI - Attaining resident duty hours compliance: the acute care nurse practitioners program at Olive View-UCLA Medical Center. AB - The institution of resident duty hours limits by the Accreditation Council for Graduate Medical Education (ACGME) has made it difficult for some programs to cover inpatient teaching services. The medical literature is replete with editorials criticizing the hour limits and the resulting problems but is nearly silent on the topic of constructive solutions to compliance. In this article, the authors describe a new program, initiated in 2003 at the Olive View-UCLA Medical Center, of using acute care nurse practitioners to allow for compliance with the "24 + 6" continuous duty hours limit, as well as the 80-hour workweek limit. Each post-call team is assigned a nurse practitioner for the day, allowing residents to sign out by 2 pm while ensuring quality care for patients. Nurse practitioners participate in evaluation of residents and, in turn, are evaluated by them. Using this system, the authors report 99% compliance with ACGME work-hour restrictions, with average work hours for inpatient ward residents decreasing from 84 to 76 hours per week. Physician satisfaction with the new system is high; anonymous evaluation by residents and faculty returned average scores of 8.8 out of 9 possible points. The authors report that using nurse practitioners on post-call days provides excellent, continuous patient care without impinging on scheduling and without sacrificing responsibility, continuity, or education for the residents. This system has several potential advantages over previously described work-hour solutions. Addition of a nurse practitioner to the post-call team is an effective solution to the problem of compliance with resident duty hours limitations. PMID- 17122463 TI - Complying with ACGME resident duty hours restrictions: restructuring the 80-hour workweek to enhance education and patient safety at Texas A&M/Scott & White Memorial Hospital. AB - Compliance with the Accreditation Council for Graduate Medical Education resident duty hours rules has created unique educational and patient-care challenges for the general medicine inpatient teaching (GMIT) teams at Texas A&M/Scott & White Memorial Hospital, including multiple patient hand-offs, multiple resident absences during teaching time, and loss of continuity of care for individual patients, all of which may have compromised patient safety. The Texas A&M/Scott & White Memorial Hospital internal medicine residency program initially complied with the duty hours rules by having residents take call every fourth night, followed by a six-hour post-call day. This system proved to be inefficient because it significantly disrupted patient care and resident education. Residents reported that this call system frequently caused them to approach the 80-hour limit and that they had difficulty leaving post-call because of unfulfilled responsibilities. They also reported sleep interruption and inadequate time to prepare for and attend educational conferences.After determining the peak admission times at the hospital, program leaders designed a call system during which the primary call team takes admissions from 12:00 pm to 8:00 pm each day, then leaves by 10:00 pm and returns after 10 hours for a full post-call day. After-hours admissions are managed by hospitalists. The solution did require hiring additional hospitalists for night-call coverage. The new structure has greatly improved the residents' experience on the GMIT teams. The entire team works together on call and post-call. Rounds and inpatient teaching continue normally on post-call days. Residents attend clinics and conferences post-call. Hand-offs are reduced greatly, and residents report that they are better rested. Residents also state that the new call system significantly enhances their education, patient care, and personal life. PMID- 17122464 TI - Training on the clock: family medicine residency directors' responses to resident duty hours reform. AB - PURPOSE: The Accreditation Council for Graduate Medical Education's 2003 restrictions on resident duty hours (RDH) raised concerns among educators about potential negative impacts on residents' training. In the early wake of these restrictions, little is known about how RDH reform impacts training in primary care. The authors surveyed family medicine (FM) residency program directors (PDs) for their perceptions of the impact of RDH regulations on training in primary care. METHOD: All PDs of 472 FM residency programs were asked via list-serve to complete an anonymous Internet-based survey in the fall of 2004. The survey solicited PDs' opinions about changes in staff and in residents' training experiences with respect to implementation of RDH regulations. Descriptive and qualitative analyses were conducted. RESULTS: There were 369 partial and 328 complete responses, for a response rate of 69% (328/472). Effects of the RDH regulations are varied. Fifty percent of FMPDs report increased patient-care duties for attendings, whereas 42% report no increase. Nearly 80% of programs hired no additional staff. Sixty percent of programs eliminated postcall clinics, and nearly 40% implemented a night-float system. Administrative hassles and losses of professionalism, educational opportunity, and continuity of care were common concerns, but a sizeable minority feel that residents will be better off under the new regulations. CONCLUSIONS: Many FMPDs cited increased faculty burden and the risk of lower-quality educational experiences for their trainees. Innovations for increasing the effectiveness of teaching may ultimately compensate for lost educational time. If not, alternatives such as extending the length of residency must be considered. PMID- 17122465 TI - The impact of resident duty hours reform on the internal medicine core clerkship: results from the clerkship directors in internal medicine survey. AB - PURPOSE: In July 2003, resident duty hours regulations were implemented. The impact of these regulations on medical student education has received minimal attention. The objective of this study was to evaluate the perceptions of internal medicine clerkship directors about the impact of resident physician duty hours reform on medical student teaching, assessment, and clerkship structure. METHOD: A survey was sent to 114 institutional members of Clerkship Directors in Internal Medicine in May 2004. The survey included 17 attitude items rated on a 5 point Likert scale, five items related to clerkship structure, and four open ended questions. Descriptive statistics were performed on the responses. RESULTS: Ninety-six surveys were returned (84%). The majority of respondents did not believe duty hours reform had a positive impact on clerkship students' educational experiences, whereas 48.3% agreed or strongly agreed that residents had more difficulty evaluating students' clinical skills. There was not a significant change in inpatient clerkship structure after duty hours implementation. Time for teaching students, concerns about a shift-work mentality, and student continuity with their teams were major challenges. Impact on ambulatory internal medicine rotations was minimal. CONCLUSIONS: Internal medicine clerkship directors are concerned about the impact of resident duty hours reform on student education. Additional studies of this educational impact are needed. PMID- 17122467 TI - Duty hours reforms in the United States, France, and Canada: is it time to refocus our attention on education? AB - Resident duty hours restrictions have now been instituted in many countries worldwide. Such policies have resulted in a broad-based discussion in the medical literature concerning their effects on patient care, resident education, and resident well-being. To better understand the impetuses behind these changes, the authors examine not only the duty hours mandates currently in effect in the United States, Canada, and France, but also the events influencing their independent development in these three countries. In the United States, an 80 hour resident workweek was mandated by the Accreditation Council for Graduate Medical Education out of concern for patient safety. In France, a 52.5-hour workweek was decreed by the government, reflecting the broader European Working Time Directive initiated out of concern for the negative impact of extended work hours on its population. In Canada, resident unions, whose primary interest has been one of resident well-being, have negotiated a series of reduced resident duty hours that approach those mandated in the United States. At the core of these changes are unique differences in these countries' health care and medical education systems. The resulting diversity in the origin and nature of such regulations serves to highlight the lack of evidence that has guided their development and the need to refocus on the educational elements of postgraduate training. PMID- 17122468 TI - Internal medicine and general surgery residents' attitudes about the ACGME duty hours regulations: a multicenter study. AB - PURPOSE: To assess internal medicine and general surgery residents' attitudes about the effects of the Accreditation Council for Graduate Medical Education duty hours regulations on medical errors, quality of patient care, and residency experiences. METHOD: In 2005, the authors surveyed 200 residents who trained both before and after duty hours reform at six residency programs (three internal medicine, three general surgery) at five academic medical centers in the United States. Residents' attitudes about the effects of the duty hours regulations on the quality of patient care, residency education, and quality of life were measured using a survey instrument containing 19 Likert scale questions on a scale of 1 to 5. Survey responses were compared using the Student's t-test. RESULTS: The response rate was 80% (159 residents). Residents reported that whereas fatigue-related errors decreased slightly, errors related to reduced continuity of care significantly increased. Additionally, duty hours regulations somewhat decreased opportunities for formal education, bedside learning, and procedures, but there was no consensus that graduates would be less well trained after duty hours reform. Residents, particularly surgical trainees, reported improvements in quality of life and reduced burnout. CONCLUSIONS: Residents in medicine and surgery had similar opinions about the effects of duty hours reform, including improved quality of life. However, resident opinions suggest that reduced fatigue-related errors have been offset by errors related to decreased continuity of care and that the quality of the educational experience may have declined. Quantifying the degree to which regulating duty hours affected errors related to discontinuity of care should be a focus of future research. PMID- 17122470 TI - The educational impact of ACGME limits on resident and fellow duty hours: a pre post survey study. AB - PURPOSE: To assess the educational impact of Accreditation Council for Graduate Medical Education resident work-hour limits implemented in July 2003. METHOD: All trainees in all 76 accredited programs at two large teaching hospitals were surveyed between May and June 2003 (before work-hour reductions) and then between May and June 2004 (after work-hour reductions) about hours, education, and fatigue. Based on changes in weekly duty hours, 13 programs experiencing substantial reduction in hours were classified into a reduced-hours group. Differences in assessments of educational endpoints before and after policy implementation by trainees in the reduced-hours group were compared with those in other programs to control for potential temporal trends, using two-way ANOVA with interaction. RESULTS: The number of respondents was 1,770 (60% response rate). The reduced-hours group reported a significant decrease in time spent directly caring for patients (from 48.5 to 42.3 mean h/wk, P = 0.03), but the volume of important clinical experiences, including procedures, was preserved, as was the sense of clinical preparedness. On 22 questions related to educational quality and adequacy, only three differences in differences were significant, with the reduced-hours group reporting a relative increase in opportunities for research, decrease in quality of faculty teaching, and decrease in educational satisfaction. The percentage of trainees reporting frequent negative effects of fatigue dropped more in the reduced-hours programs than in the other programs (P < 0.05). CONCLUSION: This study shows that it may be possible to reduce residents' hours--and the perceived adverse impact of fatigue--while generally preserving the self-assessed quality, quantity, and outcomes of graduate medical education. PMID- 17122471 TI - Medical education in Japan. AB - There are 79 medical schools in Japan--42 national, 8 prefectural (i.e., founded by a local government), and 29 private--representing approximately one school for every 1.6 million people. Undergraduate medical education is six years long, typically consisting of four years of preclinical education and then two years of clinical education. High school graduates are eligible to enter medical school. In 36 schools, college graduates are offered admission, but they account for fewer than 10% of the available positions. There were 46,800 medical students in 2006; 32.8% were women. Since 1990, Japanese medical education has undergone significant changes, with some medical schools implementing integrated curricula, problem-based learning tutorials, and clinical clerkships. A model core curriculum was proposed by the government in 2001 that outlined a core structure for undergraduate medical education, with 1,218 specific behavioral objectives. A nationwide common achievement test was instituted in 2005; students must pass this test to qualify for preclinical medical education. It is similar to the United States Medical Licensing Examination step 1, although the Japanese test is not a licensing examination. The National Examination for Physicians is a 500 item examination that is administered once a year. In 2006, 8,602 applicants took the examination, and 7,742 of them (90.0%) passed. A new law requires postgraduate training for two years after graduation. Residents are paid reasonably, and the work hours are limited to 40 hours a week. In 2004, a matching system was started; the match rate was 95.6% (46.2% for the university hospitals and 49.4% for other teaching hospitals). Sustained and meaningful change in Japanese medical education is continuing. PMID- 17122472 TI - Learning about clinical uncertainty. PMID- 17122473 TI - Challenges and issues in medical education in India. AB - The Indian medical education system, one of the largest in the world, produces many physicians who emigrate to the United States, the United Kingdom, and several other countries. The quality of these physicians, therefore, has a broad global impact. Medical schools in India have rapidly proliferated in the past 25 years, doubling since 1980 for a current total of 258. Accreditation by the Medical Council of India (MCI) emphasizes documentation of infrastructure and resources and does not include self-study. The number of schools is determined by each state; the allocation of income-generating "payment seats" in private medical schools, coupled with the high emigration, may be motivating the increase in physician production. Student selection is almost exclusively based on performance on an entrance examination, with a lower cutoff score for underrepresented minorities. Curriculum reform has been advocated for over 30 years, with calls for greater relevance of the curriculum to the needs of the community. Revised guidelines from the MCI in 1997 supported these changes. The internship year (the fifth year, focusing on rotating clinical experiences), under the aegis of medical schools in India, has suffered from lack of supervision and minimal assessment; it is often used predominantly as a time to study for residency entrance examinations. The authors recommend wider use of the in-depth accreditation process used by the National Accreditation and Assessment Council, currently applied to only 10% of medical schools, as well as reforms in curriculum, student selection, and internship assessment, in addition to stronger faculty-development efforts. PMID- 17122474 TI - The programs and context of medical education in Argentina. AB - There are 29 medical schools in Argentina (this number has increased rapidly in the last decade) offering a 6-year curriculum that usually consists of 3 years of basic science, 2 years of clinical sciences, and one internship year. Annually, 5,000 physicians graduate from these programs. Admission requirements vary depending on each university's policy. Some do not have entry requirements; others require a course, usually on the basics of mathematics, biology, chemistry or physics, and some introduction to social and humanistic studies. Each year, there are approximately 12,000 first-year medical students attending the 29 schools, which suffer a high dropout rate during the first years because of vocational problems or inability to adapt to university life. Some schools have massive classes (over 2,000 students), which makes it difficult for the schools to perfect their teaching. The number of full-time faculty members is low, and some of them have appointments at more than one medical school. Residency programs offer an insufficient number of places, and fewer than 50% of the graduates can obtain a residency position because of strict admission requirements. Coordination between the Ministry of Health, representing the health care system, and the Ministry of Education, representing the medical education system, needs to be improved. Despite the problems of medical education in Argentina, the movement to improve the education of health care workers is growing. The author offers two recommendations to help accomplish this goal. PMID- 17122475 TI - The Arabian Gulf University College of Medicine and Medical Sciences: a successful model of a multinational medical school. AB - In the late 1970s, leaders of the Arabian [corrected] Gulf countries proposed a novel idea of a joint educational and cultural venture: establishing a new regional university based in the Kingdom of Bahrain that would be managed as a multinational consortium of Gulf countries including Saudi Arabia, United Arab Emirates, Kuwait, Oman, Qatar, and Bahrain. It was intended to promote higher education and research in the Gulf region; to serve the development needs of the region; to reflect the unique economic, social, and cultural attributes of the Gulf communities and their environments; and to respond to the health care needs of the member countries. Since its inception in 1982, the College of Medicine and Medical Sciences (CMMS) at Arabian Gulf University (AGU) has adopted the educational philosophy of problem-based learning (PBL) and self-directed, student centered education. The curriculum is integrated, with early introduction of education to foster clinical skills and professional competencies. The strategic alliance with the health care systems in Bahrain and other Gulf regions has created a successful model of efficient and effective initialization of health care resources in the community. The experience that has accumulated at the AGU CMMS from introducing innovative medical education has allowed it to take a leadership position in medical education in the Gulf region. The original goals of this unique experiment have been realized along with unanticipated outcomes of spearheading changes in medical education in the Gulf region. Old and new medical schools have adopted several characteristics of the AGU educational program. Several elements contributed to its success: a clear vision of providing quality medical education and realizing and sustaining this vision by a supportive leadership at the university and college levels; an alliance with the regional health care systems; a dedicated faculty who have been able to work as a team while continually developing themselves; proper student selection and the creation of a culture of student/faculty partnerships in education and in building an international reputation and credibility by cooperating with reputable international universities and organizations. PMID- 17122476 TI - The Israel Center for Medical Simulation: a paradigm for cultural change in medical education. AB - Simulation-based medical education (SBME) is a rapidly growing field, as is illustrated by the increased development of simulation centers worldwide. SBME is becoming a powerful force in addressing the need to increase patient safety through quality-care training. Recognizing the benefits of SBME, increasing numbers of bodies involved in medical and health care education and training are establishing simulation centers worldwide. The general model of most facilities focuses on a single simulation modality or a specific branch of medicine or health care, limiting their overall impact on patient safety and quality of care across the health care systems. MSR, the Israel Center for Medical Simulation, is a comprehensive, national, multimodality, multidisciplinary medical simulation center dedicated to enhancing hands-on medical education, performance assessment, patient safety, and quality of care by improving clinical and communication skills. The center uses an "error-driven" educational approach, which recognizes that errors provide an opportunity to create a unique beneficial learning experience. The authors present the Israeli experience as an alternative model, and describe the impact of the MSR model on the Israeli medical community during four years of activity. They also describe the opportunities this model has opened towards changing the culture of medical education and patient safety within Israel Although this model may require modification when implemented in other medical systems, it highlights important lessons regarding the power of SBME in triggering and bringing about cultural changes in traditional medical education. PMID- 17122478 TI - Natural and traditional medicine in Cuba: lessons for U.S. medical education. AB - The Institute of Medicine's (IOM's) Academy of Science has recommended that medical schools incorporate information on CAM (complementary and alternative medicine) into required medical school curricula so that graduates will be able to competently advise their patients in the use of CAM. The report states a need to study models of systems that integrate CAM and allopathic medicine. The authors present Cuba's health care system as one such model and describe how CAM (or natural and traditional medicine) is integrated into all levels of clinical care and medical education in Cuba. The authors examine the Cuban medical school curriculum in which students, residents, and practicing physicians are oriented in the two paradigms of CAM and allopathic medicine. Only health professionals are permitted to practice CAM in Cuba; therefore, Cuba's medical education curriculum incorporates not only teaching about CAM, but it also teaches basic CAM approaches and clinical skills. Both the theory and practice of CAM are integrated into courses throughout the six-year curriculum. Similarities and differences between the U.S. and Cuban approaches to CAM are examined, including issues of access and cost, and levels of acceptance by the medical profession and by the public at large in both countries. The authors conclude that there is potentially much to learn from the Cuban experience to inform U.S. medical educators and institutions in their endeavors to comply with the IOM recommendations and to incorporate CAM into medical school curricula. PMID- 17122480 TI - Medicine and the arts. Mortal Lessons: Notes on the Art of Surgery [excerpt] by Richard Selzer. Commentary. PMID- 17122481 TI - Gold Foundation essay. The Francis A. Velay Humanism in Medicine essay contest: first-place essay. PMID- 17122482 TI - A cross-cultural pediatric residency program merger. AB - Academic health centers, health systems, and, to a lesser degree, medical schools and residency programs have merged, consolidated, or formed strategic alliances. There are few published reports of residency program mergers, and only one involving a merger between a historically black college and university (HBCU) and a predominantly white institution.This case study describes a merger between two dissimilar urban pediatric residency programs. The Howard University Hospital (HUH) pediatric residency program is affiliated with a HBCU, and the Children's National Medical Center (CNMC) pediatric residency program, is affiliated with a leading children's hospital which had relatively few underrepresented minority physicians or residents. The pediatric residency program merger between HUH and CNMC occurred in 2003 and presented organizational, cultural, and programmatic challenges and opportunities for both institutions. However, there was a sharp contrast between the opinions of the HUH and CNMC residents with respect to the perceived effect of the merger on residency training, patient care, and the individual institutions. Increasing the size and diversity of CNMC's resident pool and the granting of accreditation for the community health track were positive outcomes, but the magnitude of the institutional change process and the disruption to residents' routines and schedules were significant challenges. The merger served as an impetus to embed cultural competency guiding principles and expectations into the organizational fabric of the combined residency program. PMID- 17122484 TI - Developing an institution-based assessment of resident communication and interpersonal skills. AB - PURPOSE: The authors describe the development and validation of an institution wide, cross-specialty assessment of residents' communication and interpersonal skills, including related components of patient care and professionalism. METHOD: Residency program faculty, the department of medical education, and the Clinical Performance Center at the University of Illinois at Chicago College of Medicine collaborated to develop six standardized patient-based clinical simulations. The standardized patients rated the residents' performance. The assessment was piloted in 2003 for internal medicine and family medicine and was subsequently adapted for other specialties, including surgery, pediatrics, obstetrics gynecology, and neurology. We present validity evidence based on the content, internal structure, relationship to other variables, feasibility, acceptability, and impact of the 2003 assessment. RESULTS: Seventy-nine internal medicine and family medicine residents participated in the initial administration of the assessment. A factor analysis of the 18 communication scale items resulted in two factors interpretable as "communication" and "interpersonal skills." Median internal consistency of the scale (coefficient alpha) was 0.91. Generalizability of the assessment ranged from 0.57 to 0.82 across specialties. Case-specific items provided information about group-level deficiencies. Cost of the assessment was about $250 per resident. Once the initial cases had been developed and piloted, they could be adapted for other specialties with minimal additional effort, at a cost saving of about $1,000 per program. CONCLUSION: Centrally developed, institution-wide competency assessment uses resources efficiently to relieve individual programs of the need to "reinvent the wheel" and provides program directors and residents with useful information for individual and programmatic review. PMID- 17122485 TI - The predicament of osteopathic postdoctoral education. AB - The growth of colleges of osteopathic medicine (COMs) during the past 20 years has been a catalyst for change and has created new challenges in osteopathic medicine. None of these challenges is more daunting than the task of sustaining an osteopathic graduate medical education (OGME) system that has suffered during this period of rapid development. Notable trends within the osteopathic medicine community since 1990 include allopathic residency programs obtaining OGME accreditation, COM graduates bypassing OGME, repeated major changes in American Osteopathic Association (AOA) accreditation policies, a growing dependence on Accreditation Council for Graduate Medical Education programs to train osteopathic graduates, and a lessening of options for the AOA to effectively direct its OGME system. The predicament is whether COMs can continue to grow without resulting in the demise of the OGME system and a loss of professional identity. PMID- 17122489 TI - Distinction of benign sebaceous proliferations from sebaceous carcinomas by immunohistochemistry. AB - Sebaceous lesions, including sebaceous hyperplasia, sebaceomas, and sebaceous adenomas and carcinomas, are histologically distinctive adnexal proliferations with a spectrum of biological behavior ranging from benign to frankly malignant. The histologic distinction between sebaceous adenomas and carcinomas may be challenging, especially in cases showing atypical features and in small or partial biopsies. We studied multiple oncogenic and therapeutic related proteins by immunohistochemistry to identify differences in expression between benign and malignant sebaceous proliferations. A total of 27 cases, including 9 sebaceous adenomas, 4 sebaceomas, 8 sebaceous carcinomas, and 6 cases of sebaceous hyperplasia, were examined by immunohistochemistry, with antibodies directed against Ki-67 (MIB-1), bcl-2, p53, p21WAF1, p27Kip1, c-erbB-2 (Her-2/neu), CD117 (c-kit), cyclin D1, MDM2, CD99, MLH-1, and MSH-2. We found that sebaceous adenomas and sebaceomas stained like sebaceous hyperplasia did, whereas carcinomas had statistically significantly increased levels of p53 (50% versus 11%, respectively) and Ki-67 (30% versus 10%). The carcinomas also had significantly reduced levels of bcl-2 (7% versus 56%, respectively) and p21 (16% versus 34%) compared to the adenomas. Thus, a combination of several of these markers may be diagnostically useful in challenging cases. In addition, we found little or no Her-2/neu and CD117 staining, indicating that immunotherapy with Herceptin or Gleevac would likely not be useful for sebaceous carcinomas. Moreover, these results show that sebaceous adenomas and carcinomas are distinct neoplasms and provide no support for the theory that all sebaceous adenomas are truly malignant. PMID- 17122490 TI - A clinicopathological and immunohistochemical comparison of squamous cell carcinoma arising in scars versus nonscar SCC in Japanese patients. AB - Squamous cell carcinoma (SCC) of the skin shows an indolent prognosis in general. However, the prognosis of SCC arising in a scar (scar carcinoma) is considered to be worse than that of SCC without any clinical history of injury (nonscar SCC). The aim of this study was to compare several indices, p53, Ki-67, E-cadherin, and beta-catenin, which are related to tumor behavior, between scar carcinoma and nonscar SCC clinicopathologically and immunohistochemically. The materials were from 10 cases of scar carcinoma and 10 cases of nonscar SCC. Clinicopathologically, the mean ages at diagnosis of scar carcinoma and nonscar SCC were 59.2 and 71.2, respectively. The most frequent anatomic site of scar carcinoma was the limbs. The most common cause of scars in our study was burns. The mean duration from the initial injury to the diagnosis of carcinoma was 30.5 years. Immunohistochemically, the mean labeling index (calculated as the percentage of positive cells) of p53 was 16.5 and 58.6 in scar carcinoma and nonscar SCC, respectively (P < 0.01, Welch test). The LI of Ki-67 was 19.1 in scar carcinoma and 52.1 in nonscar SCC (P < 0.01, Welch test). The rates of positivity of the other proteins, such as E-cadherin and beta-catenin, were similar between scar carcinoma and nonscar SCC. In this study, the follow-up time was short and the number of patients was small, and for these reasons it might not have been possible to obtain evidence that scar carcinoma is aggressive. PMID- 17122491 TI - Comparative study of pagetoid dyskeratosis between acrochordons and soft fibromas. AB - BACKGROUND: Pagetoid dyskeratosis (PD) is considered a casual finding. We can find it in some conditions, including acrochordons and soft fibromas. OBJECTIVE: (1) to compare the presence of PD in soft fibromas and acrochordons and (2) to compare PD positive fibromas and PD negative fibromas. MATERIALS AND METHODS: We reviewed all acrochordons and soft fibromas diagnosed in the General Hospital of Lanzarote, Spain, between January 2001 and December 2002. We assessed the presence of PD, size, acanthosis, basal pigmentation, and the presence of pseudohorn cysts. RESULTS: Three hundred sixty one acrochordons and 164 soft fibromas were included in this study. There were striking differences in the presence of PD, size, acanthosis, and basal pigmentation between both entities. PD positive fibromas predominated in axillas. There were no other differences between PD positive fibromas and PD negative fibromas. CONCLUSION: Although soft fibromas and acrochordons are actually fibroepithelial polyps, including the presence of PD, there are striking differences between them. Thus, both conditions have to be considered as different entities. PD could be related to friction and moisture. PD has to be distinguished from other conditions such as Paget's disease, pagetoid melanoma, koilocytes, clear cell papulosis, among others. PMID- 17122492 TI - Photodermatitis with minimal inflammatory infiltrate: clinical inflammatory conditions with discordant histologic findings. AB - Dermatoses associated with cutaneous photosensitivity are a group of photodistributed skin eruptions caused or exacerbated by light. Multiple clinical variants of photosensitive dermatoses have been characterized including polymorphous light eruption, chronic actinic dermatitis, solar urticaria, phototoxic and photoallergic dermatitis, reticular erythematous mucinosis, acute cutaneous lupus erythematosus, and dermatomyositis. As there may be significant overlap among the clinical presentation of these conditions, the specific diagnosis of individual photodermatosis relies heavily on characteristic histopathologic features. We present here 5 cases of photodistributed eruptions with virtual absence of histologic epidermal changes and dermal inflammation, yet all were described clinically as being "inflammatory" and erythematous. All cases of this "pauci-inflammatory photodermatitis" presented with photodistributed bright red macular erythema or slightly indurated plaques that developed over a period of weeks to months and clinically resembled photoallergic or phototoxic drug reactions or polymorphous light eruption. Microscopically, however, only very sparse dermal lymphocytic infiltrate was noted with no or minimal epidermal changes. To our knowledge, the observation of clinically evident photodistributed dermatoses that demonstrate such minimal histopathologic findings has not been reported. Clinicians and histologists should be aware of the disparity that may be encountered in this setting, as the clinical features are usually far more impressive than those seen histologically. PMID- 17122493 TI - Cutaneous vasculitis update: small vessel neutrophilic vasculitis syndromes. AB - A broad and diverse spectrum of vasculitic syndromes exists. These syndromes affect the skin with varying levels of associated systemic manifestations, running the gamut from a self-limited, localized, cutaneous phenomenon to rapidly progressive, multiorgan disease. The majority of cases of cutaneous vasculitis will show a neutrophilic small vessel vasculitis that can be either a primary (idiopathic) disorder (eg, cutaneous leukocytoclastic angiitis) or a secondary disorder that is associated with drugs, infection (eg, streptococcal infection, viral hepatitis), or underlying disease (eg, connective tissue disease, malignancy). Biopsy is the gold standard for the diagnosis of cutaneous vasculitis and also necessary for the detection of cutaneous vascular immune complexes by direct immunofluorescence. Based on the type of vessel disrupted by inflammation (small and/or muscular), the distribution of vasculitis in the dermis and subcutis, and predominate inflammatory cell-type mediating vessel wall damage, a list of relevant differential diagnoses can be generated. This histologic information coupled with extravascular findings such as tissue eosinophilia, tissue neutrophilia, and/or granulomas, plus pathophysiologic markers such as direct immunofluorescent examination for immune complexes and serologic evaluation for antineutrophil cytoplasmic antibodies allows for more accurate diagnosis of specific vasculitic entities. Herein, we review both primary and secondary vasculitic syndromes that affect the skin and show a small vessel neutrophilic mediated vasculitis. PMID- 17122494 TI - Trichoblastic infundibular cyst. AB - We report 4 cases of an infundibular cyst that histopathologically showed small papillary projections of follicular germinative cells emanating from the basal layer of the cyst wall. Some of these projections branched and formed a reticulated pattern. This cystic lesion is histopathologically distinctive. We propose to designate it as a trichoblastic infundibular cyst because its lining resembles that of an infundibular cyst, yet it has cells that resemble those of the follicular germ in its wall. PMID- 17122495 TI - Follicular germinative cells in pilomatricoma. AB - Pilomatricoma is a follicular neoplasm that shows differentiation towards the follicular matrix, as characterized by matrical cells and shadow cells. In addition, this is sometimes accompanied by differentiation toward the follicular infundibulum and outer root sheath. These finding suggest that pilomatricoma can subtly differentiate towards any structure of the follicle. However, morphologically, no distinct germinative cells resembling embryonic follicular germ cells have been found in pilomatricoma to date. We report on two cases of pilomatricoma in which distinctive follicular germinative cells palisaded at the edges of collections of matrical cells. We propose two explanations: one is that the metrical cells of pilomatricoma can focally de-differentiate to form follicular germinative cells, and the other is that these follicular germinative cells are the precursor cells of pilomatricoma. PMID- 17122496 TI - Angiolymphoid hyperplasia with eosinophilia showing prominent granulomatous and fibrotic reaction: a morphological and immunohistochemical study. AB - Angiolymphoid hyperplasia with eosinophilia is an uncommon condition that usually presents in early to middle-aged adults as multiple red nodules or plaques, most commonly on the head and neck. It can also occur on the trunk and limbs, and rarely at other sites such as the breast and oral mucosa. Although one-third of these lesions recur, surgical excision is curative. The histopathological hallmark is the presence of blood vessels with plump epithelioid endothelial cells. The nonvascular component consists of histiocytes, plasma cells, eosinophils, and lymphocytes. The occurrence of numerous giant cells in the stroma of angiolymphoid hyperplasia with eosinophilia is extremely unusual and such a finding in association with diffuse granulomatous/fibrous reaction has not been reported. We describe such a lesion that developed in the wrist of a 34-year old pregnant female. PMID- 17122497 TI - Microcystic adnexal carcinoma with mandibular bone marrow involvement: a case report with immunohistochemistry. AB - Microcystic adnexal carcinoma is a rare, locally aggressive cutaneous neoplasm with a high probability of persistence locally but a low probability of metastasis. We report a case of a 69-year-old female patient with an indurated plaque at the mental region. Histologically, the tumor cells invaded the subcutaneous tissue and mandibular bone. The tumor consisted mainly of squamous and basaloid epithelial nests and cords embedded in a desmoplastic stroma. A few keratin-filled microcysts and ductal structures were also observed. Perineural encroachment was also noted but there was no mitosis, cytologic features of malignancy, or metastasis. The epithelial nests were positive to various cytokeratins except for CK20 and the lumina of the ductal structures were positive to carcinoembryonic antigen. Our results indicate that microcystic adnexal carcinoma consists of tumor cells capable of both follicular and eccrine differentiation. It is locally aggressive, extends far beyond its clinical presentation and may involve the bone. It may persist and remain asymptomatic for so many years without metastasis. A lifetime postsurgery monitoring is mandatory to ensure early and proper management. PMID- 17122498 TI - Mixed nonseminomatous germ cell tumor presenting as a subcutaneous tissue mass. AB - Extragonadal germ cell tumors most commonly arise in the midline of the retroperitoneum or the mediastinum. Primary tumors involving the skin are very rare. Only one case of malignant primary germ cell tumor located in the skin has been reported. We present the case of a 44-year-old white man with a primary subcutaneous mixed nonseminomatous germ cell tumor. This man had a long-lasting subcutaneous lump of the breast, which became painful. Surgery revealed 3 juxtaposed nodules. Microscopic examination showed a mixed germ cell tumor with a 90% immature teratoma component and a 10% embryonal carcinoma component. Testicular ultrasound and computed tomography of the chest, abdomen, pelvis, and brain were normal. Serum human chorionic gonadotrophin, beta-human chorionic gonadotrophin, alpha-fetoprotein, and lactate dehydrogenase were within normal ranges. A further surgical excision was performed. The patient is presently alive with no evidence of disease after a follow-up of 7 years. Review of the literature indicates that primary cutaneous extragonadal germ cell tumors usually occur as cutaneous or subcutaneous solitary nodules or as ulcerated lesions. They mainly consist of mature teratomas in children. Only 2 cases have been reported in adults. PMID- 17122499 TI - The men behind the eponym--Abraham Buschke and Ludwig Lowenstein: giant condyloma (Buschke-Loewenstein). AB - In 1925 Abraham Buschke and Ludwig Loewenstein described a neoplasm of the penis (carcinomahnliche condylomata acuminata des penis), which, to them, bore resemblances to common condyloma acuminatum and squamous cell carcinoma, but it had histopathologic and clinical characteristics that differed from these 2 proliferations. Later Lauren V. Ackerman described a similar neoplasm of the oral mucosa that he termed "verrucous carcinoma." I have previously reviewed that neoplasm, now referred to as "oral verrucous carcinoma of Ackerman." Still later Aird and his colleagues described a unique neoplasm of the foot, "carcinoma cuniculatum." These 3 have been grouped generically as "verrucous carcinoma." Verrucous carcinomas occur at many sites including the female genitalia and the aerodigestive system. This report will synopsize the lives of Abraham Buschke and Ludwig Loewenstein, review their original articles, and mention the changing concepts over time concerning condylomata, particularly large condylomata. Some other reports of giant condyloma (verrucous carcinoma of the penis) will be discussed, emphasizing the histopathology of this neoplasm and its differentiation from "warty-like" carcinomas of the penis. Verrucous carcinoma of other sites and epithelioma (carcinoma) cuniculatum will not be discussed. PMID- 17122500 TI - Nomenclature for very superficial squamous cell carcinoma of the skin and of the cervix: a critique in historical perspective. AB - Squamous-cell carcinoma is the most common of all cancers and it develops in diverse organs of the body, among those being the skin, lung, gastrointestinal tract, and genitourinary tract, the latter including the cervix. Unfortunately, no unanimity exists for naming very superficial squamous-cell carcinoma; it has not been designated in consistent fashion in a single organ, let alone in all of them, thereby resulting in confusion, not only in regard to terminology per se, but concerning matters conceptual, not the least of those being what appellation to apply to that condition when it is encountered histopathologically. This vexing situation is illustrated graphically in the skin by diagnoses for very superficial squamous-cell carcinoma as disparate as solar keratosis (actinic keratosis, senile keratosis), arsenical keratosis, radiation keratosis, Bowen disease, bowenoid papulosis, squamous-cell carcinoma in situ, as well as variations on the theme of "keratinocytic intraepidermal neoplasia" and "dysplasia," and in the cervix by squamous-cell carcinoma in situ, leukoplakia, cervical intraepithelial neoplasia I-III, as well as variations on the theme of "squamous dysplasia ()." What follows now is a recounting of the history of the subject under consideration here, a critique of dizzying, opaque terms and phrases given to that subject, and a proposal for rectifying what currently is a thoroughly untenable situation because the language, and the ideas expressed by it, are impenetrable to physicians and, thereby, are decidedly disadvantageous to patients. There is a need urgently for a single term for very superficial squamous-cell carcinoma in every organ of the body in which it develops, to wit, one that conveys diagnosis in such logical, lucid, comprehensible fashion that it is understandable, readily and immediately, to clinicians. In that way, physicians charged with management of patients can plan therapy rationally. PMID- 17122501 TI - Res ipse loquitur! PMID- 17122503 TI - Re: Through a lens darkly. PMID- 17122504 TI - Comparison of microsatellite instability, CpG island methylation phenotype, BRAF and KRAS status in serrated polyps and traditional adenomas indicates separate pathways to distinct colorectal carcinoma end points. AB - The aim of this study was to compare BRAF and KRAS, CpG island methylator phenotype (CIMP), and microsatellite instability (MSI) status in each of the histologic categories, including end-point carcinomas with residual adenoma, of the serrated polyp neoplasia pathway and the traditional (nonserrated) adenoma carcinoma sequence. Deoxyribonucleic acid (DNA) was extracted from the selected samples and assayed for BRAF, KRAS2 codon12, 13, CIMP using markers hMLH1, MGMT, MINT1, MINT2, p16, and MSI using an assay for BAT25 and BAT26. A BRAF mutation was present in 82% of serrated carcinomas (SCas), 62% of serrated adenomas (SAs), 83% of serrated polyps with abnormal proliferation (SPAPs-syn. sessile serrated adenoma [SSA]), 76% of microvesicular serrated polyps (MVSPs), and was not found in any of the histologic categories of the traditional adenoma-carcinoma sequence. KRAS2 mutations were found in 43% of the goblet cell serrated polyp (GCSP) category, 13% of MVSPs, 7% of SPAPs, and 24% of SAs; in 26% of large traditional adenoma (lTAs) compared with small traditional adenomas (sTAs) (0/30; P<0.005) and in 37.3% of traditional carcinomas (TCa). CIMP-H (>1 marker positive) was significantly more frequent in SPAP, SA, and SCa compared with MVSP (P<0.05); CIMP-H was present in 10% of sTAs but was found more frequently in lTA (44.4%; OR 7.2; P=0.007) and TCa (38.9%; OR 5.8; P=0.007). Higher CIMP levels (4 or more markers positive) were significantly more frequent in advanced categories of the serrated pathway (SAs [31%] and SCas [30%]) compared with lTAs [0%] and TCAs [3.4%] (OR 12.2; P=0.02). MSI-H was identified only in the adenocarcinoma component of SCas (9/11) or in the contiguous SAs (3/7). The findings indicate that a BRAF mutation is a specific marker for a serrated polyp pathway that has its origin in a hyperplastic polyp (MVSP) and a potential end point as MSI carcinoma. CIMP-High (CIMP-H) develops early in this sequence and MSI-H develops late. The data provided a less complete picture of a second serrated pathway, identified by a KRAS2 mutation in SAs, but showed that the progressive stages of both iterations of the serrated neoplasia pathway are separate and distinct from those of the traditional adenoma-carcinoma sequence. PMID- 17122505 TI - Inflammatory myofibroblastic tumors of the urinary tract: a clinicopathologic study of 46 cases, including a malignant example inflammatory fibrosarcoma and a subset associated with high-grade urothelial carcinoma. AB - Inflammatory myofibroblastic tumor (IMT) of the urinary tract, also termed postoperative spindle cell nodule, inflammatory pseudotumor, and pseudosarcomatous fibromyxoid tumor, is rare and in the past was believed to reflect diverse entities. We reviewed a series of 46 IMTs arising in the ureter, bladder, and prostate, derived primarily from a large consultation practice. There were 30 male and 16 females aged 3 to 89 years (mean 53.6). Lesions were 1.2 to 12 cm (mean 4.2). There was a history of recent prior instrumentation in 8 cases. Morphology was similar to that previously described for IMT occurring in this region, with the exception of 1 case that focally appeared sarcomatous. Polypoid cystitis coexisted in 5 patients (11%). Mitoses were typically scant (0 to 20/10 hpf, mean 1). Necrosis was seen in 14 (30%) cases. Invasion of the muscularis propria was documented in 19 (41%). By immunohistochemistry (IHC), lesions at least focally expressed anaplastic lymphoma kinase (ALK) (20/35, 57%), AE1/3 (25/34, 73%), CAM5.2 (10/15, 67%), CK18 (6/6, 100%), actin (23/25, 92%), desmin (15/19, 79%), calponin (6/7, 86%), caldesmon (4/7, 57%, rare cells), p53 (10/13, 77%), and most lacked S100 (0/14), CD34 (0/13), CD117 (2/13, 15%), CD21 (0/5), and CD23 (0/3). ALK gene alterations were detected by fluorescence in situ hybridization (FISH) in 13/18 (72%) tested cases, including 2 with prior instrumentation; 13/18 (72%) showed agreement between FISH ALK results and ALK protein results by IHC. Most bladder IMTs were managed locally, but partial cystectomy was performed as the initial management in 7 cases and cystectomy in 1 (1 IMT was initially misinterpreted as carcinoma, 1 IMT was found incidentally as a separate lesion in a cystectomy specimen performed for urothelial carcinoma). Follow-up was available in 32 cases (range 3 to 120 mo; mean 33; median 24). There were 10 patients with recurrences (2 with 2 recurrences). Recurrences were unassociated with muscle invasion or with ALK alterations. In 2 cases, tumors of the urinary tract (TURs) showing IMT preceded (1 and 2 mo, respectively) TURs showing sarcomatoid carcinoma with high-grade invasive urothelial carcinoma accompanied with separate fragments of IMT. Even on re-review the IMT in these 2 cases were morphologically indistinguishable from other cases of IMT, with FISH demonstrating ALK alterations in the IMT areas in one of them. Both these patients died of their carcinomas. Lastly, there was 1 tumor with many morphological features of IMT and an ALK rearrangement, yet overtly sarcomatous. This case arose postirradiation for prostate cancer 4 years before the development of the lesion, with tumor recurrence at 4 months and death from intra abdominal metastatic disease at 9 months. In summary, urinary tract IMTs are rare and share many features with counterparts in other sites, displaying similar morphology and immunogenotypic features whether de novo or postinstrumentation. Typical IMTs can be locally aggressive, sometimes requiring radical surgical resection, but none of our typical cases metastasized, although they can rarely arise contemporaneously with sarcomatoid urothelial carcinomas. For these reasons, close follow-up is warranted. PMID- 17122506 TI - The distribution of low and high-risk HPV types in vulvar and vaginal intraepithelial neoplasia (VIN and VaIN). AB - It has been proposed that low-grade vulvar and vaginal lesions (VIN 1 and VaIN 1) are flat condylomas and should be designated as such. Moreover, their relationship to high-grade lesions (VIN 3 and VaIN 3) is unclear. Accordingly, this study was undertaken to address these issues by comparing the distribution of human papillomavirus (HPV) types in vulvar and vaginal intraepithelial lesions. We identified 33 cases of VIN 1, 34 cases of VIN 3, 17 cases of VaIN 1, and 16 cases of VaIN 3. In addition, 36 cases of low-grade squamous intraepithelial lesion (LSIL) in the cervix and 116 cases of cervical high-grade squamous intraepithelial lesion were used for comparison. Polymerase chain reaction analysis was performed using both the Roche PGMY and DDL SPF 10 systems. In cases where HPV was detected, the majority of low-grade and high-grade lesions contained a single HPV type. However, a minority of cases were found to have multiple HPV types. Of the VIN 1 cases, a low-risk virus was seen in 22 (67%), with HPV 6 or 11 accounting for 14 (42%). A high-risk virus was detected in 14 (42%) of cases of which 2 (6%) contained HPV 16. Of the VIN 3 cases, all had high risk HPV of which 31 (91%) were found to have HPV 16. Of the VaIN 1 cases, 6 (35%) were found to have low-risk HPV types. HPV 6 or 11 were not found in these cases. High-risk virus was seen in 13 (76%) VaIN 1 cases, with 1 (6%) containing HPV 16. HPV was detected in 15 of 16 (94%) VaIN 3 lesions, all of which had high risk types. HPV 16 was found in 8 (50%). In contrast, 2 (6%) of cervical LSIL had low-risk HPV (HPV 6 and 11), whereas 34 (94%) of LSIL cases had high-risk HPVs. Of the cervical high-grade squamous intraepithelial lesion cases, 100% had high risk HPVs of which 87 (75%) were found to have HPV 16. The findings demonstrate that a significant number of low-grade vulvar and vaginal lesions contain high risk HPV types, supporting their designation as low-grade intraepithelial lesions rather than flat condylomas. The low frequency of HPV 16 in VIN 1 compared with VIN 3 suggests they are distinct lesions or that HPV 16 is critical in the progression to VIN 3. Finally, comparison of the distribution of HPV in the vagina and vulva suggests that VaIN is more closely related to cervical intraepithelial neoplasia than to VIN. PMID- 17122507 TI - Precursors of endometrial clear cell carcinoma. AB - The recognition of morphologically identifiable lesions which may confer an increased risk for subsequent development of an invasive malignancy offers an opportunity to investigate and better understand the molecular-genetic etiopathogenesis of the well-developed tumor, and potentially, to administer a therapeutic intervention before its development. In contrast to uterine endometrioid and serous carcinomas, very little is known about the potential precursor lesions of endometrial clear cell carcinoma (ECCC). In our routine practice, we have noted the presence of a spectrum of atypical glandular changes in the endometria adjacent to ECCC or endometrial carcinomas with a clear cell component, which on the basis of current criteria, would not qualify for any specific designation. We hypothesize that these lesions represent the earliest morphologically recognizable precursor lesions of ECCC and systematically characterize their clinicopathologic features herein. Thirty archived cases of pure ECCC (n=14) or mixed endometrial carcinomas with a >10% clear cell component (n=16) were retrieved and the "normal" endometria adjacent to the malignancies were evaluated in detail. Thirty-eight benign uteri and 30 uteri with classic endometrial endometrioid carcinoma (EEC) served as controls. All cases were reviewed in a blinded fashion. Putative precursor lesions (PPL) were searched for and identified microscopically. The lesions were typically isolated glands or surface epithelium (within an otherwise normal endometrial region) that displayed cytoplasmic clarity and/or eosinophilia with varying degrees of nuclear atypia. Twenty-seven (90%) of the 30 cases had at least 1 PPL. In contrast, PPL were identified neither in the benign uteri nor in endometrioid carcinoma control groups (P<0.001). A total of 67 foci of PPL were identified in the 27 cases with an average of 2.5 foci per case. The immunohistochemical expression of p53, mib 1, estrogen receptor (ERs), and progesterone receptor in the benign endometria, ECCC, and the PPL were evaluated on all 27 cases. The mean p53 scores for the benign endometria, PPL, and ECCC were 0, 4.5, and 6.2, respectively. Parallel values for mib-1 were 15%, 45%, and 63%. ER/progesterone receptor indices for benign endometria, PPL, and carcinoma were 90/80, 21.52/4.61, and 11/4, respectively. The PPL described herein have a morphologic and immunophenotypic profile which seems to be distinct from both the benign endometria in which they reside and the adjacent areas of ECCC. The high frequency of association of these lesions with ECCC, their frequent occurrence as isolated lesions within otherwise "benign-appearing" endometria, and their continuous spectrum of nuclear atypia from minimum (grade 1, cytologic atypia falls short of ECCC cells) to maximum (grade 3, cytologically identical to ECCC cells), argues in favor of our hypothesis that these may represent precursor lesions of ECCC. Further studies are required to conclusively define the nature of these lesions. However, such studies can only be performed if diagnostic surgical pathologists recognize, highlight, and segregate these lesions for further analysis. PMID- 17122508 TI - Microcarcinoids in large intestinal adenomas. AB - Composite adenoma-carcinoid tumors are rare colorectal lesions consisting of intermingled adenomatous and carcinoid components. Unlike other mixed endocrine glandular colorectal neoplasms, which are generally malignant, their glandular component is histologically benign and their natural history is favorable. We present 4 cases of colonic adenomas containing microcarcinoids, a hitherto undescribed lesion that is either a precursor of composite adenoma-carcinoids or a related but independent entity. The cases, identified among our surgical and consultation files, were endoscopically routine sessile polyps removed from 4 otherwise normal individuals, 3 from the cecum and 1 from the distal colon. The microcarcinoids were 0.5 to 1.5 mm in size and situated within the basal lamina propria, where they interposed between the crypts and muscularis mucosae without disturbing the overall polyp architecture. Histologically, they consisted of collections of low-grade epithelial cells arranged in nests, cords, tubules, and irregular clusters and characterized by eosinophilic, granular, or clear cytoplasm and by round central nuclei with stippled or dusty chromatin. Endocrine differentiation of the microcarcinoids was confirmed by the expression of 3 or more of the following: Grimelius argyrophil, chromogranin, synaptophysin, neuron specific enolase and somatostatin. No mitotic figures or MIB-1 or p53 positivity were observed. The glandular component of the polyps was unremarkable in 3 cases, but 1 polyp, in addition to a microcarcinoid, showed a diffuse pattern of mixed adenomatous-endocrine differentiation. The patients' clinical course was benign on the basis of 2 years' median follow-up (range, 6 mo to 10 y). Two patients with incomplete polypectomies underwent hemicolectomy revealing no residual endocrine neoplasia. Awareness of microcarcinoids in colonic adenomas should help avert potential diagnostic pitfalls posed by their pleomorphism, basal location, and infiltrative patterns, and may help clarify their natural history and possible relationship to composite glandular-carcinoid tumors. PMID- 17122509 TI - Autoimmune pancreatitis: a systemic immune complex mediated disease. AB - Autoimmune pancreatitis (AIP) is a mass forming inflammatory pancreatobiliary centric disease. Recent reports of multiorgan inflammatory mass forming lesions with increased numbers of IgG4 positive plasma cells suggest that AIP may have a systemic component. In this study, we explore the systemic nature of AIP, investigate the relevance of subtyping AIP, perform a systematic study of tissue IgG4 immunoperoxidase, and ultrastructurally evaluate the presence of immune complexes. Our study group consisted of 36 patients with AIP, 21 of whom underwent a Whipple procedure. On the basis of the pattern of inflammation, pancreatic involvement was subtyped as ductocentric (AIP-D) or lobulocentric (AIP L). Extrapancreatic lesions included bile duct (n=3), salivary glands (n=3), lung (n=2), gallbladder (n=11), and kidney (n=4). Clinical and radiologic data was recorded. Immunohistochemistry for IgG4 was performed on both pancreatic and extrapancreatic tissues and the numbers of IgG4 positive plasma cells were semiquantitatively scored. A control cohort composed of pancreatic adenocarcinoma (n=19) and chronic pancreatitis-not otherwise specified (NOS) (n=14) was also evaluated. Eleven pancreatic specimens, including 2 cases of chronic pancreatitis NOS and 4 kidneys were evaluated ultrastructurally. The pancreas, bile duct, gall bladder, salivary gland, kidney, and lung lesions were characterized by dense lymphoplasmacytic infiltrates with reactive fibroblasts and venulitis. IgG4 positive plasma cells were identified in all pancreatic and extrapancreatic lesions. The AIP cases showed significantly more pancreatic IgG4 positive plasma cells than chronic pancreatitis-NOS or adenocarcinoma (P=0.001). However, IgG4 positive cells were identified in 57.1% of chronic pancreatitis-NOS and 47.4% of ductal adenocarcinoma. Fifteen of 21 resected cases were classified as AIP-D, and 6 as AIP-L, the latter notably showing significantly more IgG4 positive plasma cells than the former (P=0.02). Additionally, clinical and radiologic differences emerged between the 2 groups. Ultrastructurally, electron dense deposits of immune complexes were identified in the basement membranes of 7 of the 9 AIP cases and in 3 of the 4 renal biopsies evaluated. AIP represents the pancreatic manifestation of a systemic autoimmune disease. Clinical and immunologic findings justify the recognition of pancreatic lobulocentric and ductocentric subtypes. Documentation of increased numbers of tissue IgG4 positive plasma cells, although not an entirely specific marker for AIP, may provide ancillary evidence for the diagnosis of a IgG4-related systemic disease. PMID- 17122510 TI - The incidence and anatomic site specificity of chromosomal translocations in primary extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) in North America. AB - Several balanced translocations have been identified in extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) but there are few data regarding their frequency in different anatomic sites or the frequency of translocations involving BCL6 or kappa or lambda immunoglobulin light chain genes (IGK or IGL), particularly in patients from geographic regions other than Europe and Japan. One hundred thirty-three paraffin-embedded North American primary MALT lymphoma specimens from diverse anatomic sites were studied by fluorescence in situ hybridization (FISH) using probes for API2-MALT1, IGH MALT1, IGH-BCL10, IGH-FOXP1, IGH, +/- centromeres 3, 7, 12, and 18, and a subset (n=74) were analyzed using FISH probes for IGK, IGL, and BCL6. Translocations were mutually exclusive and were detected in 26% of cases (17% API2-MALT1, 5% IGH MALT1, 3% IGH-unknown translocation partner, and 1% IGH-BCL10). Aneuploidy was associated with IGH-MALT1 and IGH-BCL10 but only rarely with API2-MALT1. There was striking site specificity, with API2-MALT1 showing a marked predilection for lung and intestine, and IGH-MALT1 and IGH-BCL10 occurring almost exclusively in lung. Twenty-three percent of translocation-negative primary MALT lymphomas from diverse sites showed complete/partial trisomy 18. No MALT lymphomas with translocations involving IGK, IGL, BCL6, or FOXP1 were identified. This FISH panel detected cytogenetic abnormalities in half of all MALT lymphomas, and translocations arose preferentially in MALT lymphomas of the lung and gastrointestinal tract. Differences in incidence and anatomic site specificity of translocations between North American and non-North American cases may reflect geographic variability of infectious or other etiologic factors. PMID- 17122511 TI - Expanding the histologic spectrum of mucinous tubular and spindle cell carcinoma of the kidney. AB - Mucinous tubular and spindle cell carcinomas (MTSCs) are polymorphic neoplasms characterized by small, elongated tubules lined by cuboidal cells and/or cords of spindled cells separated by pale mucinous stroma. Nonclassic morphologic variants and features of MTSC have not been well studied. We identified 17 previously unreported MTSCs from Surgical Pathology and consultative files of the authors and their respective institutions and studied their morphologic features. A total of 10/17 cases were considered "classic," as described above, with 5/10 showing at least focal (20% to 50%) tubular predominance without apparent mucinous matrix. Alcian blue staining revealed abundant (>50%) mucin in all classic cases. Seven of 17 MTSCs were classified as "mucin-poor," with little to no extracellular mucin appreciable by hematoxylin and eosin. Four of these cases showed equal tubular and spindled morphology, 2 cases showed spindle cell predominance (70%; 95%), and 1 case showed tubular predominance (90%). In 5/7 mucin-poor cases, staining for Alcian blue revealed scant (<10%) mucin in cellular areas with the other 2 cases having 30% mucin. Unusual histologic features identified in the 17 cases were: foamy macrophages (n=8), papillations/well formed papillae (n=6/n=1), focal clear cells in tubules (n=3), necrosis (n=3), oncocytic tubules (n=2; 40%, 5%), numerous small vacuoles (n=2), heterotopic bone (n=1), psammomatous calcification (n=1), and nodular growth with lymphocytic cuffing (n=1). An exceptional case contained a well-circumscribed, HMB45-positive angiomyolipoma within the MTSC. MTSCs may be "mucin-poor" and show a marked predominance of either of its principal morphologic components, which coupled with the presence of other unusual features such as clear cells, papillations, foamy macrophages, and necrosis, may mimic other forms of renal cell carcinoma. Pathologists must be aware of the spectrum of histologic findings within MTSCs to ensure their accurate diagnosis. PMID- 17122512 TI - Intraductal papillary mucinous neoplasm (IPMN) of the pancreas: its histopathologic difference between 2 major types. AB - Intraductal papillary mucinous neoplasm (IPMN) is a unique pancreatic neoplasm developing in the ductal system. Two major histologic subtypes have been reported, that is the gastric type and the intestinal type. However, their histopathologic features, especially those of the gastric type, have not been fully described. To evaluate the features of these two types and refine their differences, we analyzed 80 IPMNs including 50 cases of the gastric type and 30 cases of the intestinal type with mucin immunohistochemistry. By defining a main duct-type lesion as predominantly involving the main pancreatic duct with or without branch ducts, and a branch duct-type lesion as exclusively centered on branch ducts or consisting of a collection of small cystic lesions, gastric-type IPMNs were mostly branch duct-type lesions (98%), whereas the intestinal-type IPMNs were usually main duct type (73%). The histologic grade of the intestinal type was generally higher than that of the gastric type. The intestinal type was also characterized by frequent intraluminal nodular growth, and severe atrophy and fibrosis of the surrounding parenchyma with mucous lake formation. In contrast, pyloric glandlike structures at the base of the papillae and pancreatic intraepithelial neoplasia (PanIN)-like complexes were more frequently observed in the gastric type. A significant difference was observed between the gastric type and the intestinal type with regard to all the above features (P<0.05). Seven cases (23%) of the intestinal type were associated with an invasive adenocarcinoma (6 mucinous and 1 ductal), versus only 1 case (2%) of the gastric type (invasive ductal carcinoma). All cases of both gastric and intestinal types expressed MUC5AC; however, high immunolabeling scores for MUC2 were mostly observed in the intestinal type (P<0.05). In conclusion, gastric and intestinal types of IPMNs have distinct histopathologic features and mucin profiles, suggesting that they may follow different biologic pathways. PMID- 17122513 TI - Glypican 3: a novel marker in testicular germ cell tumors. AB - Glypican 3 (GPC3), a membrane-bound heparin sulfate proteoglycan, may play a role in promoting embryonic cell growth and differentiation. GPC3 is mutated in Simpson-Golabi-Behmel syndrome, characterized by tissue overgrowth and an increased risk of embryonal malignancies. Recently, GPC3 was reported to be one of the over-expressed genes in testicular yolk sac tumors by gene expression microarray analysis. However, the presence of the GPC3 protein in germ cell tumors has never been investigated. The purpose of the study was to investigate the GPC3 expression in various histologic components of testicular germ cell tumors using immunohistochemistry and to assess its possible utility as a diagnostic marker. Tumors from 71 patients were examined, including components of 42 seminomas, 37 embryonal carcinomas, 24 yolk sac tumors, 20 teratomas with mature elements, 16 teratomas with immature elements, and 7 choriocarcinomas. Cytoplasmic and membranous immunoreactivity was semiquantitatively evaluated. All yolk sac tumor (24/24) and choriocarcinoma (7/7) components were immunoreactive for GPC3, whereas only 38% of teratomas with immature elements and 8% of embryonal carcinomas expressed GPC3. There was no immunoreactivity in benign testicular tissue, intratubular germ cell neoplasia, seminomas (0/42), or teratomas with mature elements (0/20). We conclude that the oncofetal protein GPC3 is a novel immunohistochemical marker in testicular germ cell tumors with differential expression in defined histologic subtypes. Our findings suggest a possible role of GPC3 in tumor cell differentiation. Furthermore, GPC immunostaining may be useful in the pathologic diagnosis of nonseminomatous germ cell tumors, particularly yolk sac tumor, and choriocarcinoma. PMID- 17122514 TI - Tumor suppressor gene alterations in respiratory epithelial adenomatoid hamartoma (REAH): comparison to sinonasal adenocarcinoma and inflamed sinonasal mucosa. AB - Respiratory epithelial adenomatoid hamartoma (REAH) is an unusual benign sinonasal glandular proliferation. REAH is not considered a neoplasm, although, no molecular evidence exists to support or refute this possibility. Microdissection of 10 cases of REAH, 9 cases of sinonasal adenocarcinoma (SNAC) and 10 cases of chronic sinusitis was performed. DNA was extracted and polymerase chain reaction performed using fluorescently labeled primers flanking known tumor suppressor genes on chromosomes 9p (CDKN2/p16), 11p (H-ras), 17p (p53), and 18q (DCC/DPC4). Polymerase chain reaction products were analyzed semiquantitatively by capillary electrophoresis. Allele ratios were calculated using the peak height from the shorter allele divided by the peak height from the longer allele. The loss of heterozygosity (LOH) ratio was calculated as the allele ratio from tumor tissue divided by the allele ratio from normal tissue. The fractional allelic loss (FAL) was calculated as the percentage of loci that harbored LOH divided by the number of loci that were informative. REAH demonstrated an intermediate FAL of 31% compared with SNAC (64%) and chronic sinusitis (2%). REAH and SNAC had the highest LOH for multiple loci located on 9p (p16) and 18q (DCC/DPC4). The molecular profile of REAH shows a mean FAL of 31%, which would be considered unusually high for a non-neoplastic entity. Appreciable allelic loss within REAH suggests the possibility that REAH may be a benign neoplasm rather than a hamartoma. PMID- 17122515 TI - Sporadic duodenal bulb gastrin-cell tumors: association with Helicobacter pylori gastritis and long-term use of proton pump inhibitors. AB - We reviewed the clinicopathologic profile of a series of recently diagnosed sporadic duodenal gastrin-cell (G-cell) tumors. All cases were discovered incidentally and had a unique clinicopathologic profile: all 18 cases were gastrin-positive tumors located in the duodenal bulb, were small in size (mean size 5.4 mm), demonstrated an insular architectural pattern, and were localized to the lamina propria and submucosa. None of the patients had Zollinger-Ellison or carcinoid syndrome. The behavior was indolent and there was no evidence of metastasis at diagnosis or during follow-up. In our sampled population, the presence of Helicobacter pylori gastritis and the use of proton pump inhibitors (PPIs) were significantly associated with the presence of G-cell tumors. Both the presence of H. pylori gastritis and use of PPI remained significant in a logistic regression model adjusted for age, race/ethnicity, and sex with P values of 0.0016 (odds ratio=10.1, 95% confidence interval: 2.3 to 42.4) and 0.008 (odds ratio=8.9, 95% confidence interval: 1.76 to 45.4), respectively. Most patients with tumors showed G-cell hyperplasia in the nontumorous regions of the duodenum. The high incidence of sporadic duodenal G-cell tumors in patients with H. pylori gastritis and long-term PPI use suggests an association that needs to be further explored. Presence of G-cell hyperplasia in the nontumorous duodenal mucosa suggests that these may originate from a proliferative phase, similar to the hyperplasia-dysplasia-neoplasia sequence seen in other endocrine tumors. PMID- 17122516 TI - The predictive value of CK19 and CD99 in pancreatic endocrine tumors. AB - Prediction of behavior in pancreatic endocrine tumors (PETs) is reliant on clinicopathologic features. However, there remains a cohort of PETs that behave aggressively despite showing indolent pathologic features. Recently, it has been suggested that CK19 and CD99 are sensitive ancillary markers that predict outcome in PETs. An analysis of 54 PETs and 2 resected liver metastases was undertaken to examine the relationship of CK19 and CD99 and the pathologic criteria in the WHO classification of PETs. CK19 was found to correlate with mitotic count (>5/50 high-power fields), an MIB-1 labeling index of > or =2%, lymphovascular/perineural permeation, lymph node involvement, and liver spread. Although not statistically significant, CK19-negative tumors tended to be smaller than the average tumor size in the series (2.5 vs. 3.6 cm). CD99 did not show any significant correlation with any of the WHO criteria. Tumors that are confined to the pancreas with low mitotic count and MIB-1 labeling index, tended to be CD99 positive. Both CK 19 (negative) and CD99 (positive) correlated with insulin positive PETs. In conclusion, CK 19 may prove to be a useful ancillary diagnostic test in the routine work-up of PETs. CD99 does not appear to be as useful. There is no compelling evidence, from our study, to suggest that both these markers may be used in concert to predict the behavior of PETs. PMID- 17122517 TI - Nuclear beta-catenin accumulation as reliable marker for the differentiation between cystic craniopharyngiomas and rathke cleft cysts: a clinico-pathologic approach. AB - Clinical and histopathologic differentiation of cystic lesions from the sellar region, that is, craniopharyngiomas (CPs) and Rathke cleft cysts (RCCs), is challenging and has paramount importance with respect to variable clinical manifestation and adapted surgical treatment strategies in both entities. Here, we retrospectively evaluated clinico-pathologic findings in 81 patients presenting with a cystic tumor located in the sellar region. All patients underwent transsphenoidal or transcranial resections. Microscopic inspection of surgical specimens identified CP in 51 patients, and RCC in 30 patients. Amongst the panel of immunohistochemical marker proteins used for histopathologic analysis, nuclear accumulation of beta-catenin was detectable only in CP. On the basis of the histopathologic and immunohistochemical analysis, clinical presentation (sex, age, ophthalmologic, and endocrinologic deficits), imaging (tumor location, size, and calcification), as well as a description of cyst contents obtained during operation were retrospectively evaluated. In purely cystic CPs, an isointense signal was more frequent in T1-weighted magnetic resonance images and calcification of the tumor capsule in computed tomography scans. In addition, the size of RCC was smaller and this tumor entity was more often located within the sella. Aberrant (nuclear) immunohistochemical staining for beta-catenin appeared, however, as most reliable factor for the differentiation between purely cystic CPs and RCCs, whereas tumor location, tumor size, and calcification of the tumor capsule were less consistent parameters. The data are compatible with distinct pathogenic pathways associated with these related histopathologic entities. PMID- 17122518 TI - Impact of Epstein-Barr virus in monomorphic B-cell posttransplant lymphoproliferative disorders: a histogenetic study. AB - The heterogeneity of the posttransplant lymphoproliferative disorders (PTLDs) is well recognized. However, in contrast to other immunodeficiency-associated lymphomas or diffuse large B-cell lymphomas in general, studies of the histogenetic spectrum of the large category of monomorphic B-cell cases have been more limited, produced conflicting results, and have paid little attention to the impact of Epstein-Barr virus (EBV). Therefore, 30 monomorphic B-cell PTLD from 27 patients were analyzed using EBER in situ hybridization for EBV and a panel of antibodies directed against CD20, CD3/bcl-6, CD10, MUM-1/IRF4, CD138, and bcl-2. The results were correlated with the histopathologic features and clinical outcome. All PTLD were CD20 with 23% CD10, 53% bcl-6, 67% MUM-1/IRF4, 13% CD138, 83% bcl-2 and 67% EBV. 30% of the PTLD had a germinal center (GC) profile (CD10, bcl-6, MUM-1/IRF4, CD138), 53% a "late GC/early post-GC" profile (CD10, bcl-6, MUM-1/IRF4, CD138), 13% a post-GC profile (CD10, bcl-6, MUM-1/IRF4, CD138) and 3% an indeterminate profile (all markers negative). EBV positivity was associated with MUM-1/IRF4 expression (P=0.005) and with a non-GC phenotype (P=0.01). All CD138 cases were EBV. The cases with a GC phenotype were the most likely to resemble transformed GC cells (P=0.023). No statistically significant survival differences could be documented between those with a GC versus non-GC phenotype. These results highlight the broad histogenetic spectrum of monomorphic B-cell PTLD. They demonstrate the association of EBV positivity with a non-GC phenotype and suggest that EBV PTLD are more like lymphomas that arise in immunocompetent individuals. The lack of a demonstrable correlation with survival may relate to the relatively small number of cases studied. PMID- 17122519 TI - Comparative analysis of germ cell transcription factors in CNS germinoma reveals diagnostic utility of NANOG. AB - The homeodomain transcription factor, NANOG, along with OCT3/4 (POU5F1) and SOX2, is part of the core set of transcription factors that maintain embryonic stem cell self-renewal and pluripotency. Expression of NANOG has been detected in fetal germ cells and in gonadal germ cell tumors. To assess the diagnostic utility of NANOG in central nervous system (CNS) germ cell tumors, we analyzed its expression by immunohistochemistry in a series of 12 CNS germinomas and compared its expression with other stem cell markers. Strong nuclear expression of NANOG was demonstrated in >90% of the tumor cells in all cases. In contrast, expression of OCT3/4 and placental alkaline phosphatase was inconsistent and SOX2 was expressed in only rare cells. NANOG was not detected in tumor types frequently considered in the differential diagnosis of CNS germinoma: pineoblastoma, primitive neuroectodermal tumors, medulloblastoma, lymphoma, pituitary adenoma, atypical teratoid/rhabdoid tumor, Langerhans cell histiocytosis, and gliomas. These findings demonstrate that NANOG is a sensitive and specific marker of CNS germinoma. Compared with other currently used markers, NANOG may have superior diagnostic characteristics and can facilitate identification of germinomas in minute surgical biopsies commonly obtained from these tumors. These findings also suggest a potential biologic role for NANOG in maintenance of CNS germinoma. PMID- 17122520 TI - Restoration of bile ducts in drug-induced vanishing bile duct syndrome due to zonisamide. AB - Prolonged drug-induced cholestasis may be due to destruction and disappearance of bile ducts, sometimes referred to as vanishing bile duct syndrome. Although some of these cases progress to fibrosis, cirrhosis, and liver failure, others improve with time. We report a case of a 35-year-old man who developed vanishing bile duct syndrome after ingestion of zonisamide, an antiepileptic drug that is also prescribed for weight loss. His liver biopsy showed complete absence of bile ducts 3 weeks after starting treatment. There was no ductular reaction. The drug was stopped and a follow-up biopsy 3 months later showed strands of intermediate hepatobiliary cells at the periphery of the portal tracts that extended into lobules; these structures lacked lumina and expressed biliary cytokeratins, CK7 and CK19. A third biopsy, 7 months later, showed the presence of ductules with lumina located within portal tracts. Intermediate hepatobiliary cells were rare; although sparse clusters of hepatocytes with membrane staining for CK7 were present. Cholestasis and levels of bilirubin improved over time. The histologic features in this case document the sequence of events in restoration of the biliary tree after loss of bile ducts, which seems to be a process of maturation of intermediate hepatobiliary cells that arise from a proliferative compartment at the porto-hepatic interface. PMID- 17122521 TI - Soft tissue perineurioma in a patient with neurofibromatosis type 2: a tumor not previously associated with the NF2 syndrome. AB - Neoplasms that commonly affect patients with neurofibromatosis type 2 (NF2) include schwannomas, meningiomas, astrocytomas, ependymomas, and neurofibromas. Perineuriomas are rare tumors of the peripheral nerve sheath that share some characteristics with meningioma. As in both NF2-associated and sporadic cases of schwannoma and meningioma, perineuriomas often harbor mutations or deletions of the NF2 gene. However, perineuriomas have not previously been reported in the clinical setting of NF2. A 30-year-old man with a history of bilateral vestibular schwannomas, a parasagittal meningioma, an intraspinal ependymoma, and multiple other neoplasms involving both cranial and peripheral nerves (thereby fulfilling the diagnostic criteria for NF2) presented with an enlarging thigh mass. The diagnosis of cellular soft tissue perineurioma was confirmed by both immunohistochemical and ultrastructural analysis. This case represents the first report of a soft tissue perineurioma arising in the setting of NF2. PMID- 17122523 TI - Microadenomatosis of the pancreas in von Hippel-Lindau disease. PMID- 17122524 TI - Physician specific prescribing information. PMID- 17122525 TI - Glycemic control with insulin glargine as part of an ethnically diverse, community-based diabetes management program. AB - OBJECTIVE: To evaluate the contribution of long-acting basal insulin therapy (insulin glargine) to glycemic control in a predominantly Hispanic population participating in a community-based diabetes management program, Project Dulce. RESEARCH DESIGN AND METHODS: This retrospective analysis included 3122 adult patients with diabetes from 17 community clinics in San Diego County, California who participated in Project Dulce between July 2000 and March 2003. A subset of 180 patients received insulin glargine because of ongoing, inadequate glycemic control (ie, elevated HbA1c). Glycemic control was evaluated by mean adjusted HbA1c during follow-up clinical visits using hierarchical linear modeling, with values determined separately before and after initiation of insulin glargine. RESULTS: At baseline, the mean number of individuals with hypoglycemia, presence of diabetic complications, and duration of diabetes were greater in the glargine group that in the reference group. HbA1c at baseline was 8.79 and 9.44 (P = 0.019) in the reference and glargine groups, respectively. Mean adjusted HbA1c in the glargine group was 8.80 at baseline, 7.89 before initiation of insulin glargine (P < 0.001 vs baseline), and 7.34 after adding insulin glargine (P < 0.001 vs pre-glargine). In the reference group, mean adjusted HbA1c decreased from 8.81 at baseline to 7.40 during follow-up (P < 0.001 vs baseline). CONCLUSIONS: A comprehensive program of diabetes care in Project Dulce improved HbA1c significantly in a predominantly Hispanic population. Adding long-acting basal insulin therapy with insulin glargine produced significant incremental improvement in HbA1c. PMID- 17122526 TI - Preliminary experience with the use of oral linezolid in infants for the completion of antibiotic therapy in the outpatient setting after admission to the pediatric intensive care unit. AB - Advances in medical technology have led to improved survival after catastrophic illnesses. Many of the survivors require ongoing care including tracheostomy, mechanical ventilation, tube feedings, and indwelling venous catheters. Repeated hospitalizations may be necessary to treat infectious complications resulting from resistant organisms requiring intravenous antibiotic therapy. Because prolonged intravenous access may be difficult or even impossible in these patients, alternative means of therapy are necessary. Linezolid is the first of a new class of antimicrobial agents known as the oxazolidinones with activity against gram-positive bacteria similar to that of vancomycin and yet its oral bioavailability allows for enteral administration. We present our retrospective experience with oral linezolid in a cohort of pediatric intensive care unit patients. Primary infectious disease issues included endocarditis, tracheitis, pneumonia, or central line sepsis resulting from Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus, and Enterococcus. Treatment was initiated with vancomycin and changed to enteral linezolid (10 mg/kg every 12 hours). The duration of therapy with linezolid varied from 7 days to 6 weeks. All of the patients were discharged home to complete their course of enteral linezolid. No complications related to linezolid therapy were noted, and all of the patients completed their prescribed course of therapy without the need for rehospitalization. Our preliminary experience suggests that oral linezolid offers an effective alternative to intravenous vancomycin for the treatment of infections resulting from gram-positive bacteria and avoids the need for prolonged vascular access. PMID- 17122527 TI - Analysis of the effects of fentanyl in the feline pulmonary vascular bed. AB - The objective of this study was to test the hypothesis that fentanyl induces a depressor response in the pulmonary vascular bed of the cat and to identify the receptors involved in the mediation or modulation of these effects. The authors conducted a prospective vehicle-controlled study at a university research laboratory using intact chest preparation in adult mongrel cats. In separate experiments, the effects of diphenhydramine (histamine receptor blocker), glibenclamide (ATP-sensitive K+ channel blocker), L-N5-(1-Iminoethyl) ornithine hydrochloride (L-NIO) (nitric oxide synthase inhibitor), nimesulide (selective cyclooxygenase [COX]-2 inhibitor), and naloxone (opiate receptor antagonist) were investigated on pulmonary arterial responses to fentanyl and other agonists in the pulmonary vascular bed of the cat. The systemic pressure and lobar arterial perfusion pressure were continuously monitored, electronically averaged, and recorded. In the feline pulmonary vascular bed of the isolated left lower lobe, fentanyl induced a dose-dependent vasodepressor response that was not significantly altered after administration of glibenclamide, L-NIO, and nimesulide. However, the responses to fentanyl were significantly attenuated after administration of diphenhydramine and naloxone. The results of the present study suggest that fentanyl has potent vasodepressor activity in the pulmonary vascular bed of the cat and that this response may be mediated or modulated by both histaminergic and opiate receptor sensitive pathways. PMID- 17122528 TI - Compliance with poison center fomepizole recommendations is suboptimal in cases of toxic alcohol poisoning. AB - We sought to examine hospital compliance with poison center antidotal alcohol dehydrogenase inhibition recommendations in cases of ethylene glycol (EG) and methanol (ME) ingestion. A 2-year analysis of all potential EG and ME ingestion cases reported to a regional poison center was conducted. Excluded from analysis were exposures without an ingestion, without a confirmatory EG or ME serum assay, or without complete medical charting. During the study period, 579 EG or ME exposures were reported to the poison center: 133 cases met study eligibility as an ingestion. Of the 133 cases, 102 (77%) had complete data and were included in the analysis. Immediate alcohol dehydrogenase inhibition was recommended by the poison center in 79 of the 102 cases. Fomepizole was recommended in 61/79 (77%); ethanol was recommended as an alternative therapeutic choice in 32/61 (52%) of these cases if fomepizole was not immediately available. Ethanol alone was recommended in 18/79 (23%). Fomepizole was eventually administered in 39/61 (64%) cases where recommended. The mean time to antidote administration was 3 times longer in cases where a choice in antidote was given [57 min (95% confidence interval, 43-70) vs. 146 min (95% confidence interval, 93-200)]. Despite its ease of administration, fomepizole is used less frequently than recommended by poison center staff. Delays to antidote administration occurred more commonly in cases where the poison center gave a choice in antidotal therapy. PMID- 17122529 TI - Ciclesonide disposition and metabolism: pharmacokinetics, metabolism, and excretion in the mouse, rat, rabbit, and dog. AB - The pharmacokinetics, metabolism, and excretion of ciclesonide, a novel and effective inhaled glucocorticoid for the treatment of asthma, were investigated after intravenous and oral administration of 14C-ciclesonide in the mouse, rat, rabbit, and dog. The pharmacokinetics of ciclesonide in all animal species were characterized by a low oral bioavailability (approximately 6% or less), a high clearance, and a large volume of distribution. The apparent terminal half-life of ciclesonide was short; the apparent terminal half-life of the active desisobutyryl-ciclesonide metabolite (des-CIC or M1) was longer and ranged from 2.4 to 6.9 hours in the 4 species. Metabolites derived from ciclesonide in serum (or plasma) and excreta samples from the 4 animal species were profiled and identified by LC/RAM/MS (liquid chromatography/radioactivity monitor/mass spectrometry). Ciclesonide was extensively metabolized to yield des-CIC, which was further metabolized to primarily yield hippuric acid and hydroxylated metabolites, namely, isomers of cyclohexane-monohydroxylated des-CIC and B-ring monohydroxylated des-CIC. Greater than 90% of intravenous and oral 14C ciclesonide doses were recovered in all species; the main elimination route was fecal/biliary. A comparison of in vitro and in vivo metabolite profiles between mice, rats, rabbits, and dogs with those from humans indicated that metabolic pathways for ciclesonide were qualitatively similar in humans and in the 4 animal species. PMID- 17122530 TI - Infliximab drug and infusion costs among patients with Crohn's disease in a commercially-insured setting. AB - Significant attention has been paid to the cost of treating patients with anti TNF agents for diseases including Crohn's disease (CD). To evaluate the actual expenditures for infliximab in routine clinical practice, the authors examined patterns of administration and reimbursement among commercially-insured patients receiving infliximab for CD. Integrated pharmacy and medical claims data were obtained from 78 US health plans. Data from CD patients treated with infliximab between June 2000 and December 2003 were analyzed. Each claim for an infusion (HCPCS code J1745) represented a single observation. Descriptive statistics were generated for the number of vials billed for each infusion, total charged amount (ie, the amount submitted by providers), and total paid amount (ie, the amount reimbursed by the health plan, net of any patient copayment, coinsurance, or deductible). A total of 2230 CD patients were identified; of the original total of 9724 infusions, 168 were not evaluable because of data quality issues, yielding cost evaluation for 9556 total infusions. At each infusion, claims for a mean (SD) of 4.79 (1.74) vials were submitted (median = 5). Corresponding charged amounts were 4441 dollars (1778 dollars) (median = 4099 dollars); paid amounts averaged 2793 dollars (990 dollars) (median = 2628 dollars). Charged and paid amounts per vial billed averaged 927 dollars and 583 dollars, respectively. The average cost of administering a dose of infliximab to a commercial insurer is approximately 2800 dollars. Assumptions regarding infusion costs for infliximab based on charged amounts may be misleading; the true costs of administering infliximab in routine practice are likely to be lower than that reported for charged amounts. PMID- 17122531 TI - Comparing patient telephone callback rates for different hormonal birth control delivery systems. AB - This study was conducted to determine the number of office telephone callbacks in the first 3 months after initiating use of the vaginal ring, the transdermal patch, or an oral contraceptive. If a patient called back, the reason for her call was noted. Patients were prospectively followed from sites in New Jersey and Florida for 3 months after initiation of hormonal contraception (oral, transdermal, vaginal) and the number of callbacks for each method was assessed. Before study recruitment, the patients had either never used or discontinued hormonal contraception for at least 1 month. All patients were given standard counseling, which included reference handouts specific to their chosen form of contraception. The number of callbacks for patients on oral contraceptives was 30/96 (26 patients, 4 of the 26 with 2 callbacks each); for the transdermal patch, 20/40 (17 patients, 3 of the 17 with 2 callbacks each); and for the vaginal ring, 11/72 (11 patients). Pearson chi2 analysis revealed that all 3 groups were significantly different from each other (P = 0.0004). Most patients called back for side effects related to their method of choice, and some patients called back more than once. Rate of callbacks after initiation of hormonal contraception was the least with the vaginal ring. Use of oral contraception, often considered the gold standard, resulted in a callback rate that was midway between the number of callbacks for the vaginal ring and the number of callbacks for the transdermal patch. The lower observed rate of callbacks with the vaginal ring compared with oral and transdermal contraception may provide clinicians the confidence that this method is well tolerated by their patients. PMID- 17122532 TI - Efficacy of radiofrequency catheter ablation in treatment of elderly patients with supraventricular tachyarrhythmias and ventricular tachycardia. AB - Radiofrequency catheter ablation was performed in 100 men and 81 women, mean age 78 +/- 5 years, referred for ablation of atrial flutter, supraventricular tachycardia, and ventricular tachycardia, and for ablation of the atrioventricular junction with permanent pacemaker implantation in patients with atrial fibrillation and a rapid ventricular rate not controlled by drug therapy. A hematoma in 1 of 182 ablation procedures (<1%) was the only complication. Radiofrequency catheter ablation was successful in treating 63 of 70 patients (90%) with atrial flutter, in treating 60 of 66 patients (91%) with supraventricular tachycardia, in treating 2 of 2 patients (100%) with ventricular tachycardia, and in ablating the atrioventricular junction in 43 of 44 patients (98%) with atrial fibrillation and a rapid ventricular rate not controlled by drug therapy. PMID- 17122533 TI - Alzheimer disease and its management: a review. AB - Alzheimer disease is a progressive degenerative disorder of the brain characterized by a slow, progressive decline in cognitive function and behavior. As the disease advances, persons with Alzheimer disease have tough time with daily usage of things like using the phone, cooking, handling money, or driving the car. The disease is more common in elder population. It is estimated that Alzheimer disease affects 15 million people worldwide and approximately 4 million Americans. The clinical features of Alzheimer disease overlaps with common signs of aging, and other types of dementia, hence the diagnosis remains difficult. The neuropathologic hallmarks of the disorder are amyloid-rich senile plaques, neurofibrillary tangles, and neuronal degeneration. Drugs approved for treating Alzheimer disease include acetylcholinesterase inhibitors and N-methyl-D aspartate (NMDA) receptor antagonist. Caregivers not getting adequate information about Alzheimer disease may believe that nothing can be done to manage its symptoms. Understanding the extent of Alzheimer disease related knowledge can assist disease management that result in improved disease management and reduced care costs. This article attempts to focus on some of the important recent developments in understanding and management of Alzheimer disease. PMID- 17122534 TI - Failure of enforcement controlled substance laws in health policy for prescribing opiate medications: a painful assessment of morbidity and mortality. AB - Controlled substances can be used for legitimate medical purposes to relieve pain and suffering, and allow management of medical and surgical conditions, whether acute or chronic in duration. However, because these are attractive, addicting drugs, diversion from sources such as physicians and pharmacists can lead to serious health problems. Of importance is that addiction to opiate medications can interfere with treatment of the original pain condition, and can lead to life threatening states because of poor judgment and depressed mood in the users. Consequently, the public has a vested interest in protecting the medical uses of these medications on the one hand, although reducing the morbidity and mortality from their diversion and addictive use. The controlled substance laws contain 3 sources of policy framework that governs the medical use and diversion of controlled substances: (1) international treaties, (2) federal laws and regulations, and (3) state laws and regulations. These laws are aimed at balancing the need to controlling use with adverse consequences against the therapeutic benefits opiate medications provide the public. PMID- 17122535 TI - Propoxyphene (dextropropoxyphene): a critical review of a weak opioid analgesic that should remain in antiquity. AB - Propoxyphene (dextropropoxyphene) is a synthetic weak opioid introduced into the United States in 1957. It is most frequently prescribed in combination with acetaminophen and/or aspirin. After its ubiquitous introductory phase, it was soon discovered that this drug's iatrogenic events (cardiotoxicity, seizures, etc.) far outweighed any perceived therapeutic benefit. Propoxyphene analgesia was equated with that of merely acetaminophen or aspirin independently. The propoxyphenes euphorigenic component has created a problem in its prescribing. Use of this agent in the elderly should be avoided because of its complex pharmacokinetics and pharmacodynamics. The pharmacokinetics, pharmacodynamics, and pharmacology of this drug are discussed thoroughly in this article, including its arrhythmogenicity. Additional noncardiovascular pharmacotherapies that produce QTc prolongation or arrhythmogenicity are described. A list of the cytochrome P450 2D6 pharmacotherapies that will interact with propoxyphene is provided in the article. The use of this agent is highly discouraged. The rationale for this is discussed fully within this article. The toxicity of this drug is partially related to nor-propoxyphene a non-opioid cardiotoxic metabolite. The mere warnings of fatalities within the package insert should alert any cautious prescriber on the dangers of this agent and dampen its prescribing potential. PMID- 17122536 TI - Acute chorea associated with metoclopramide use. AB - Metoclopramide is a dopamine receptor antagonist that is used to treat diabetic gastroparesis, chemotherapy-induced nausea, and migraines. It is known to cause extrapyramidal side effects such as tardive dyskinesia, parkinsonism, dystonia, and akithisia, but not chorea. We describe a patient who presented with choreiform movements shortly after the administration of intravenous metoclopramide. Her work-up for secondary causes of chorea was otherwise negative and her symptoms abated with the administration of oral quetiapine and intravenous diazepam. PMID- 17122537 TI - Methemoglobinemia-induced cardio-respiratory failure secondary to topical anesthesia. AB - Acute toxic methemoglobinemia is an infrequent complication of the use of topical anesthetics, most notably benzocaine. The clinical picture is characterized by sudden development of tissue hypoxia without underlying cardiac or respiratory dysfunction, and deceptively normal oxygen saturation on conventional arterial blood gas analysis. This condition may be rapidly fatal and management depends upon prompt recognition, confirmation of clinical suspicion using cooximetry of arterial blood, and quick institution of therapy. We describe the first reported case of cardio-respiratory failure associated with acute toxic methemoglobinemia, which was initially misconstrued as an evolving acute coronary event and rapidly responded to methylene blue therapy. PMID- 17122538 TI - Rare case of serotonin syndrome with therapeutic doses of paroxetine. AB - Paroxetine (Paxil) is a selective serotonin reuptake inhibitor (SSRI) and anxiolytic that is approved to treat numerous mood disorders. Serotonin syndrome, defined as a triad of mental status changes, autonomic instability, and neuromuscular abnormalities, is a potentially life-threatening complication of administering such serotonin-modifying drugs. Most cases of serotonin syndrome that have occurred with paroxetine administration are due to inadvertent drug interactions, most notably between SSRIs and monamine oxidase inhibitors, or intentional overdoses. The authors present the case of an 80-year-old woman who presented with serotonin syndrome while on a therapeutic dose of paroxetine. Paroxetine was stopped, and aggressive hydration with fluids and treatment with cyproheptadine was followed by remarkable improvement and return to baseline status in 4 days. This case illustrates the importance for physicians to have a heightened sense of suspicion of the serotonin syndrome in any patient known to be on serotonin-modifying agents presenting with altered sensorium and cholinergic symptoms. Consequently, they will be able to start timely treatment without subjecting the patient to unnecessary and potentially harmful tests. PMID- 17122539 TI - Severe pulmonary complications in African-American patient after bortezomib therapy. AB - Bortezomib is a proteosome inhibitor with good clinical activity in multiple myeloma. Frequently described side effects are gastrointestinal symptoms, neuropathy, and thrombocytopenia. Even though pneumonia is listed as an infrequent toxicity, severe pneumonitis leading to respiratory distress had not been described until recently. This report was from a single institution in Japan. All these patients had received bone marrow transplant before therapy with bortezomib. To the authors knowledge, this is the first report of life threatening pulmonary toxicity after bortezomib in a non-Japanese patient and without history of prior autologous peripheral stem cell transplant. PMID- 17122540 TI - Relapse of high-grade non-Hodgkin's lymphoma after autologous stem cell transplantation: a case successfully treated with cyclophosphamide plus somatostatin, bromocriptine, melatonin, retinoids, and ACTH. AB - Patients with relapse of high-grade non-Hodgkin lymphoma (NHL) after autologous stem cell transplantation (auto-SCT) generally have a poor prognosis. Only a minority of these patients can be cured by a second myeloablative chemotherapy, and conventional salvage treatments are often associated with severe toxicities. With a combination of cyclophosphamide, somatostatin, bromocriptine, retinoids, melatonin, and ACTH, we already reported 100% global response in 8 patients with relapse of low-grade NHL after single or combined chemotherapy and a therapy-free period of > or = 6 months. This provided the rationale to evaluate the same pharmacological association in a patient with relapse of high-grade NHL after auto-SCT performed 2 years before. The patient was treated for at least 2 months. At the end of this period, if he had stable or responding disease, he received additional 3 months of treatment, and if he was stable or responding after 5 month, he was treated for 3 months and more. After 2 months, patient had a partial response, and after 5 months, he achieved a complete response. Today, 14 months after beginning treatment, patient is in complete remission. Treatment had very good tolerance, and patient carried on at home doing his normal activities. Our result and severe toxicities associated with conventional salvage treatments suggest in a relapse of high-grade NHL after auto-SCT, further clinical trials using the pharmacological association we employed in this case. PMID- 17122541 TI - The superficial inferior epigastric artery (SIEA) flap: indications for breast reconstruction. AB - The lower abdominal skin and fat have become a standard for breast reconstruction because the abdominal tissue can mimic the breast to a high degree. With today's increasing focus on safety and routine of microsurgical interventions, more attention must be paid to donor-site morbidity. The superficial inferior epigastric artery (SIEA) free flap is the least invasive microsurgical procedure for breast reconstruction because this operative technique does not require harvesting of the rectus muscle or the abdominal fascia. A total of 11 breast reconstructions were performed with this technique and had an average follow-up of 23 months. The anatomy, operative technique, and various indications for the SIEA flaps are demonstrated. PMID- 17122542 TI - Latissimus dorsi muscle flap and tissue expansion for breast reconstruction. AB - BACKGROUND: After mastectomy for breast cancer, especially when combined with radiation, inadequate wall thickness and insufficient elasticity of the skin are problems frequently encountered in breast reconstruction. We describe a reconstruction method using a latissimus dorsi muscle flap, followed by expansion, that creates no additional scar. METHODS: This retrospective study included 30 patients who underwent this 4-surgery reconstruction: the muscle flap was raised and drawn through the mastectomy scar; the expander was inserted; expander replacement with the definitive prosthesis and concomitant symmetrization of the contralateral breast; finally, reconstruction of the nipple areola complex. RESULTS: Only 1 failure, prosthesis extrusion, was observed. A surgeon scored the outcomes as above average for 26 women, who were satisfied. DISCUSSION: We noted an overall lower complication rate and a lower reconstruction-failure rate compared with reported results. This approach obtained better tissue quality; the flap provided better covering thickness and expansion, yielding a larger muscle-skin pocket. CONCLUSIONS: This reconstruction procedure seems reliable and extends the indications of skin expansion, with satisfactory results. PMID- 17122543 TI - New aspects of breast volume measurement using 3-dimensional surface imaging. AB - Precise and objective calculation of breast volume is helpful to evaluate the aesthetic result of breast surgery, but traditional methods are unsatisfactory. Three-dimensional (3D) scanning of the body surface allows reproducible and objective assessment of the complex breast region but requires further investigation before clinical application. The main goal of this study was to investigate the precision and accuracy of breast volume measurement using 3D body scanning. Five independent observers standardized the 3D scanning method using 2 dummy models (n = 200) and examined its applicability with 6 test subjects and 10 clinical patients (n = 2220). Breast volume measurements obtained with the 3D scanner technology were compared with reference measurements obtained from test subjects through nuclear magnetic resonance imaging. The mean deviation of the breast volume measurements of 1 test subject by all observers, expressed as percentage of volume, was 2.86 +/- 0.98, significantly higher than the deviation for the dummy models, 1.65 +/- 0.42 (P < 0.001). With respect to all clinical patients, the mean measurement precision obtained preoperatively was less precise than that obtained postoperatively (3.31 +/- 1.02 versus 1.66 +/- 0.49, respectively). Interobserver differences in measurement precision were not statistically significant. The mean breast volumes obtained by nuclear magnetic resonance imaging (441.42 +/- 137.05 mL) and 3D scanning (452.51 +/- 141.88 mL) significantly correlated (r = 0.995, P < 0.001). Breast volume measurement with 3D surface imaging represents a sufficiently precise and accurate method to guarantee objective and exact recording. PMID- 17122544 TI - Immediate reconstruction of the nipple/areola complex in oncoplastic surgery after central quadrantectomy. AB - BACKGROUND: Oncoplastic surgery adds valuable techniques for breast-conservation therapy that allows for wide excisions and prevents breast deformities. However, no such technique has addressed loss of the nipple/areola complex (NAC) after central lumpectomy. We present a simple and effective technique for immediate reconstruction of the NAC after such loss due to tumorectomy. METHODS: After central tumorectomy, a local tissue flap is created above the defect to restore the nipple. Then the neonipple is carried on a superior-based, dermoglandular pedicle to its new position, similar to breast reduction surgery. The operation is continued with resection of redundant tissue lateral to the pedicle for optimal breast shape. From the discarded breast tissue, a full-thickness skin graft is harvested and used to reconstruct the areola. The contralateral breast is treated with a usual mammaplasty and tailored to the specific needs of the opposite side. RESULTS: Nine patients with central tumors of the breast were treated in this fashion at our institution. In all patients, the aesthetic result was good to excellent. In 1 patient, there was delayed wound healing of the full thickness skin graft for the areola, which healed by secondary intention. CONCLUSION: The presented technique is easily achieved and produced excellent results after breast-conservation surgery. It will expand the armamentarium of oncoplastic surgery to meet central defects with loss of the nipple/areola complex. PMID- 17122545 TI - Optimal management and challenges in treatment of upper facial melanoma. AB - This study's purpose was to evaluate clinical and surgical outcomes in patients with upper facial melanoma. A sentinel lymph node (SLN) biopsy database review identified 43 patients receiving a diagnosis of upper facial melanoma between February 1997 and April 2005 at Mayo Clinic Arizona in Scottsdale. Patients underwent wide local excision (n = 40) or Mohs excision (n = 3) and SLN biopsy. Nine patients (21%) had positive margins requiring reexcision. SLN mapping identified the SLN in 39 patients (91%) and drainage to bilateral lymph node basins in 8 (21%). The SLN was positive for melanoma in 2 patients (5%). Recurrence in 33 patients with more than 1 year of follow-up (local in 5 [15%] and regional in 1 [3%]) was treated with salvage surgery; 1 patient developed metastatic disease. Two patients (5%) died, one of an unknown cause and the other of metastatic melanoma. We concluded that oncologic surgery can result in good local disease control in patients with upper facial melanoma. PMID- 17122546 TI - Management of enterocutaneous fistulas using negative-pressure dressings. AB - Fifteen patients with enterocutaneous fistulas (ECFs) not amenable to surgical treatment were treated with negative-pressure dressings over the abdominal wound and ECF. Closure of the ECF and time to closure were examined. In 11 patients who had no visible intestinal mucosa on examination, the closure rate was 100%, with a mean time to closure of 14 days. In 4 patients who did have grossly visible intestinal mucosa, no closure occurred. This represents an overall closure rate of 73%. Fistula output rate did not have a significant effect on outcome. These results confirm the efficacy of negative-pressure dressings in the closure of ECFs. Presence or absence of visible intestinal mucosa is the single most important clinical factor when considering the use of a negative-pressure dressing in the management of a patient with ECF. PMID- 17122547 TI - Borderline indications for ear reconstruction. AB - While the indication for an ear reconstruction with rib cartilage is clear in pediatric patients and patients without previous surgery, there are borderline cases in which neither a reconstruction nor a prosthesis seems to be first choice. Within the last 6 years, approximately 120 patients were treated with a reconstruction with rib cartilage and 20 with a prosthesis at our hospital. Patients without extensively scarred periauricular tissue clearly benefit by a reconstruction with cartilage. The classic indication for an ear prosthesis is status post-malignant tumor resection. In borderline cases presenting with heavy scars, the most favorable repair method has to be decided upon individually. Under these circumstances, one of the main criteria for a reconstruction with rib cartilage is a viable temporoparietal fascia flap. A consideration that is important to note is that an unfavorable ear reconstruction can be converted almost always into a prosthesis repair but very rarely vice versa. PMID- 17122548 TI - Vertical distraction osteogenesis of fibular bone flap in reconstructed mandible. AB - Excellent functional and aesthetic results can be achieved in mandibular reconstructions with using free fibular bone flap. However, the vertical deficiency between the reconstructed segment and the occlusal plane made dental rehabilitation impossible in some cases. We encountered this problem in our 3 patients who had mandibular reconstruction with fibular flap due to extensive bone defect result from gunshot injury. To overcome this segmental vertical distraction of the reconstructed mandible was performed. Fibular bone segments (40-70 mm) were distracted with using extraoral distraction device after a latency period of 5-7 days. The rate of distraction was 1 mm/day, and the rhythm was 4 times (4 x 0.25 mm). Distraction was continued until the desired height was achieved, and the distractor left in place for 12 weeks for bony consolidation. No minor or major complications were encountered. The increase of vertical height was between 9 and 13 mm, and it was stable during the follow-up period (7-22 months). Following the vertical distraction and vestibuloplasty operations, the dental restoration of the patients was performed with mandibular removable partial dentures. PMID- 17122549 TI - Vascularized capitate transposition for advanced Kienbock disease: application of 40 cases and their anatomy. AB - Based on anatomic study, the vascularized capitate transposition to replace excised necrotic lunate was designed and applied in 40 cases of advanced Kienbock disease. It includes excision of the necrotic lunate and proximal shift of the vascularized capitate. The blood supply of the transposed capitate is provided by the dorsal branch of the anterior interosseous artery. Bone union occurred radiographically, and no postoperative capitate necrosis occurred in all cases after 6 weeks. Twenty-three cases were followed up for 1 year. No residual wrist pain existed in the range of motion, but limited residual wrist pain existed in labor work. The arc of motion ranged on average from 35 degrees of flexion to 45 degrees of extension. The grip power of the affected hand reached on average 70% compared with the contralateral. The authors conclude the vascularized capitate transposition is a reliable alternative for advanced Kienbock disease. PMID- 17122550 TI - Paravertebral turnover flaps for closure of large spinal defects following tethered cord repair. AB - Reconstruction of large congenital spinal defects remains a challenge. We present our experience in closure of difficult spinal defects following repair of tethered cord, using paravertebral fascial or myofascial turnover flaps. Between 1996 and 2005, 23 patients were operated using paravertebral turnover flaps for closure of large spinal defects associated with tethered cord. Fifteen (65%) patients had lipomyelomeningoceles. Eleven (48%) patients had sacral defects, 10 (43%) had lumbosacral defects, and 2 (9%) had lumbar defects. Fourteen (61%) patients underwent closure using fascial turnover flaps. Myofascial turnover flaps were used in 9 (39%) patients. Following surgery, none of the patients developed cerebrospinal fluid (CSF) leaks, pseudomeningoceles, or subcutaneous infection. One patient suffered superficial necrosis and infection of the skin suture line, which healed secondarily. We conclude that fascial or myofascial paravertebral turnover flaps provide reliable coverage of difficult defects of the spinal CNS. PMID- 17122551 TI - A new technique for correction of congenital constriction rings. AB - BACKGROUND: Congenital constriction ring (CCR) is among the most common congenital anomaly of the extremities. However, few techniques have been described for correction of this anomaly, and none of these techniques could provide a complete elimination of the deformity. Therefore, new surgical techniques are needed for correction of this challenging deformity. PURPOSE: Here, a new technique, namely, Mutaf procedure, for the correction of congenital constriction rings is described. MATERIAL AND METHOD: In this technique, following excision of the fibrotic constriction ring, the groove is filled with the turnover dermofat flaps elevated from both sides of the ring in an alternating pattern. Then skin closure is done with rectangular-plasty technique. Over 8 years, this new technique was used for correction of severe CCRs of the lower extremities in 7 patients aged 1-7 years. RESULTS: In all patients, a normal extremity contour was obtained, with complete elimination of the sandglass deformity caused by the constriction ring. A mean follow-up of 3 years revealed a normal growth of the operated extremity in all patients. The scars were found to be esthetically acceptable. No further surgery was required. CONCLUSIONS: Filling the circular groove caused by the constriction ring with dermofat flaps, this new technique eliminates the soft tissue deficiency and provides a normal extremity contour. Moreover, since rectangular-plasty allows replacing the major limbs of the incisional scars within the relaxed skin tension lines, it provides a better scar in comparison with old Z-plasty techniques. PMID- 17122552 TI - Increasing the success rate of the reversed-flow fasciocutaneous island sural flap: a clinical experience in 26 cases. AB - The reversed-flow fasciocutaneous island sural flap is presenting one of the best reconstructive options in the distal third of the leg, around the ankle and the heel up to the metatarsals. Although the surgical maneuver of this flap and many of its modifications have been well described in previous publications, the flap is unfortunately full of reported complications. In this work, we are presenting a 26-patient series of reversed-flow fasciocutaneous island flap cases in different age, sex, and etiologic groups. All flaps had survived, with a complication rate of 3.8% (1 case) in the form of partial necrosis. In this series, we standardized a modified technique in which we tried to include many ideas and tricks, in addition to some of the reported useful ones to have a higher success rate. We have summarized these points in 10 golden rules, making this flap safer and more versatile. PMID- 17122553 TI - Medial plantar artery island flap for heel reconstruction. AB - Coverage of soft tissue defects of the heel has been a challenge to reconstructive surgeons in the past. The medial plantar artery flap has facilitated heel coverage since its development in the 1980s. This was a prospective study in 2 centers assessing the complications and durability of this flap primarily in patients with sensory impairment. All patients but 1 had chronic plantar ulceration due to sensory loss, and 5 patients also had developed squamous cell carcinoma. Fifty-one flaps were carried out in 48 patients. One flap underwent necrosis, and delayed healing was seen in 4 cases. Total flap survival was 98%. Minor revision of the flap or its pedicle was required in 3 cases. With a mean follow-up of 14 months, there were recurrences of ulceration in 7 feet (14%). The relevant literature is reviewed. The medial plantar artery flap is a reliable flap for heel coverage in both our centers and others. It is durable and long lasting even in insensate. PMID- 17122554 TI - Transfer of training in robotic-assisted microvascular surgery. AB - BACKGROUND: Transfer of training refers to the ability to transfer acquired skills from one discipline to another. This study aims to determine whether experience in traditional freehand microsurgery facilitates mastery of robotic microsurgery. METHODS: Microsurgical anastomoses of coronary arteries harvested from explanted pig models were used to demonstrate whether prior experience with microsurgery is required in learning robot-assisted microsuturing. Eighty microsurgical anastomoses were performed. Three fully trained vascular surgeons (n = 3) (Group A) and 5 midlevel surgical residents (n = 5) (Group B) performed the anastomoses. Each subject performed 5 freehand and 5 robotic-assisted (Zeus robotic system) anastomoses. Anastomosis time and integrity of anastomoses were recorded, including errors of management (EOM) (breaking suture, breaking knots, breaking or damaging needles). RESULTS: For fully trained surgeons, all anastomoses in the robotic-assisted group were mechanically intact. There was significantly increased anastomosis time with the robot (Robot: 14 minutes, versus freehand: 7.2 minutes, P < 0.01). The robotic-assisted anastomoses were associated with a higher EOM (Robot: 1.2, versus freehand: 0.3, P < 0.01). Surgical trainees had longer anastomosis times with robotic assistance (Robot: 14.8 minutes, versus freehand, 12.7 minutes; P < 0.01) and increased EOM (Robot: 1.6, versus freehand: 1.0; P < 0.05).Overall, surgical trainees and fully trained vascular surgeons had longer anastomotic times with robotic assistance [Robot: 14.0 versus 14.8 minutes; P = not significant (NS)], and EOM (Robot: 1.6, versus freehand: 1.2; P = NS) were not significantly different. CONCLUSION: The technical feasibility of performing a safe and efficient robotic-assisted microsurgical anastomosis in explanted vessels was repeatedly tested and demonstrated in this study within reasonable time required for the anastomosis. Compared with conventional microanastomosis, both fully trained surgeons and residents demonstrated an ability to master the robotically assisted procedure with similarly longer anastomosis times and EOM. This study indicates that robotically assisted microanastomosis can be mastered equally well by surgical trainees and fully trained vascular surgeons. PMID- 17122555 TI - Venous thrombosis in coupled versus sutured microvascular anastomoses. AB - BACKGROUND: The anastomotic coupling device (ACD) is currently used for microsurgical venous anastomoses as an alternative to sutures and can reduce anastomotic time. METHODS: We reviewed the microsurgical procedures performed at the University of Texas M. D. Anderson Cancer Center over a 40-month period. Procedures were performed for elective reconstruction of the breast, head or neck, or extremities after tumor excision. Thrombosis and ultimate flap salvage rates were compared between cases using ACDs and those using sutures for the venous anastomoses. RESULTS: Seven hundred twenty-three free flaps were included in the study. There were no intraoperative technical complications using the ACD. Overall, the venous thrombosis rates were not significantly different between the ACD (1.4%, 2/139 cases) and sutured (3.3%, 19/584 cases) groups. Salvage rates following venous thrombosis were not significantly different for venous anastomoses performed with an ACD (50%, 1 of 2 cases salvaged) compared with venous anastomoses performed with sutures (68.4%, 13 of 19 cases salvaged). CONCLUSION: The ACD is a useful alternative to sutured venous anastomoses that can reduce anastomotic time while achieving similar patency rates. PMID- 17122556 TI - Transcutaneous electrical nerve stimulation in viability of a random skin flap in nicotine-treated rats. AB - BACKGROUND: Skin flaps are extensively used in plastic surgery, the main postoperative problems being ischemia and necrosis. Nicotine has acute vasoconstrictive properties, diminishing blood flow and increasing the risk of flap necrosis. Transcutaneous electrical nerve stimulation (TENS) was effective in reducing necrosis in random skin flaps in rats. Therefore, the authors decided to test the efficacy of TENS to reduce necrosis in the presence of nicotine. METHODS: Sixteen adult male Wistar-Escola Paulista de Medicina (EPM) rats were distributed, at random, in 2 groups of 8 animals. The nicotine group (NG) received this drug subcutaneously (1.2 mg/kg/d), for 1 week before flap elevation. The nicotine plus TENS group (NTG) followed the same routine. Immediately after the operative procedure and on the next 2 postoperative days, the treatment group (NTG) was submitted to TENS during an hour under anesthesia. NG animals were sham exposed. After 7 days, the percentages of necrosis were determined. RESULTS: The NG had a mean value of 45% and the NTG, 21% of necrosis, a difference statistically significant (P < 0.001). CONCLUSION: TENS was effective in the reduction of necrosis in this flap model. PMID- 17122557 TI - Functional reconstruction of the pelvic ring with simultaneous bilateral free fibular flaps following total sacral resection. AB - The treatment of large sacral tumors involving the pelvis is challenging and may require radical resection. Sacral resection disrupts the continuity between the spine and pelvis, resulting in loss of skeletal support for weight bearing. Without pelvic reconstruction, patients remain nonambulatory. This report describes a case of functional pelvic ring reconstruction utilizing bilateral simultaneous free fibular flaps following radical sacrectomy. Vascularized free fibular grafts may be used to restore pelvic continuity for cases involving total sacral resection. PMID- 17122558 TI - Managing extracavitary prosthetic vascular graft infections: a pathway to success. AB - Prosthetic vascular graft infections portend grave consequences if not treated expediently. Despite the low incidence of infection, the potential for limb loss or death greatly magnifies this complication. The surgical management of prosthetic graft infections has evolved over the last 2 decades. With the myriad therapeutic options now available, an algorithm is necessary to provide the optimal surgical treatment of Samson groups 1 through 5 extracavitary infected vascular prostheses. An extensive review of the literature was undertaken to evaluate the most effective management schemes. The authors found that 3 factors- Samson classification, bacteriology, and patient vascular anatomy--are vital to the surgical strategy. These 3 criteria were examined, and an algorithm was developed based on successful clinical and experimental results. This review provides a step-by-step rationale for the surgical management of extracavitary prosthetic graft infections according to the most successful reported outcomes. PMID- 17122559 TI - The old, the young, and the computer or musings of a plastic surgery curmudgeon. PMID- 17122560 TI - Invited discussion: pedicled deep inferior epigastric perforator flap: an alternative method to repair groin and scrotal defects. PMID- 17122561 TI - You need a lot of "plastic" to become a plastic surgeon. PMID- 17122565 TI - To see or not to see. PMID- 17122566 TI - Do the right thing (or Do the market exclusivity thing?). PMID- 17122567 TI - Forehead pulse oximetry: Friend and foe. PMID- 17122568 TI - In hot pursuit. PMID- 17122569 TI - Statins: The next advance in cardioprotection? PMID- 17122570 TI - Trends in anesthesia-related death and brain damage: A closed claims analysis. AB - BACKGROUND: The authors used the American Society of Anesthesiologists Closed Claims Project database to determine changes in the proportion of claims for death or permanent brain damage over a 26-yr period and to identify factors associated with the observed changes. METHODS: The Closed Claims Project is a structured evaluation of adverse outcomes from 6,894 closed anesthesia malpractice claims. Trends in the proportion of claims for death or permanent brain damage between 1975 and 2000 were analyzed. RESULTS: Claims for death or brain damage decreased between 1975 and 2000 (odds ratio, 0.95 per year; 95% confidence interval, 0.94-0.96; P < 0.01). The overall downward trend did not seem to be affected by the use of pulse oximetry and end-tidal carbon dioxide monitoring, which began in 1986. The use of these monitors increased from 6% in 1985 to 70% in 1989, and thereafter varied from 63% to 83% through the year 2000. During 1986-2000, respiratory damaging events decreased while cardiovascular damaging events increased, so that by 1992, respiratory and cardiovascular damaging events occurred in approximately the same proportion (28%), a trend that continued through 2000. CONCLUSION: The significant decrease in the proportion of claims for death or permanent brain damage from 1975 through 2000 seems to be unrelated to a marked increase in the proportion of claims where pulse oximetry and end-tidal carbon dioxide monitoring were used. After the introduction and use of these monitors, there was a significant reduction in the proportion of respiratory and an increase in the proportion of cardiovascular damaging events responsible for death or permanent brain damage. PMID- 17122571 TI - Survey of anesthesia-related mortality in France. AB - BACKGROUND: This study describes a nationwide survey that estimates the number and characteristics of anesthesia-related deaths for the year 1999. METHODS: Death certificates from the French national mortality database were selected from the International Classification of Diseases, Ninth Revision codes using a variable sampling fraction. Medical certifiers were sent a questionnaire (response rate, 97%), and the anesthesiologist in charge was offered a peer review (acceptance rate, 97%). Files were reviewed to determine the mechanism of each perioperative death and its relation to anesthesia. Mortality rates were calculated using the number of anesthetic procedures estimated from a national 1996 survey and compared with a previous (1978-1982) nationwide study. RESULTS: Among the 4,200 certificates analyzed, 256 led to a detailed evaluation. The death rates totally or partially related to anesthesia for 1999 were 0.69 in 100,000 (95% confidence interval, 0.22-1.2 in 100,000) and 4.7 in 100,000 (3.1 6.3 in 100,000), respectively. The death rate increased from 0.4 to 55 in 100,000 for American Society of Anesthesiologists physical status I and IV patients, respectively. Rates increased with increasing age. Although concerns regarding aspiration of gastric contents remain, intraoperative hypotension and anemia associated with postoperative ischemic complications were the associated factors most often encountered. Deviations from standard practice and organizational failure were often found to be associated with death. CONCLUSION: In comparison with data from a previous nationwide study (1978-1982), the anesthesia-related mortality rate in France seems to be reduced 10-fold in 1999. Much remains to be done to improve compliance of physicians to standard practice and to improve the anesthetic system process. PMID- 17122572 TI - A phase I, two-center study of the pharmacokinetics and pharmacodynamics of dexmedetomidine in children. AB - BACKGROUND: To investigate dexmedetomidine in children, the authors performed an open-label study of the pharmacokinetics and pharmacodynamics of dexmedetomidine. METHODS: Thirty-six children were assigned to three groups; 24 received dexmedetomidine and 12 received no drug. Three doses of dexmedetomidine, 2, 4, and 6 microg x kg x h, were infused for 10 min. Cardiorespiratory responses and sedation were recorded for 24 h. Plasma concentrations of dexmedetomidine were collected for 24 h and analyzed. Pharmacokinetic variables were determined using nonlinear mixed effects modeling (NONMEM program). Cardiorespiratory responses were analyzed. RESULTS: Thirty-six children completed the study. There was an apparent difference in the pharmacokinetics between Canadian and South African children. The derived volumes and clearances in the Canadian children were V1 = 0.81 l/kg, V2 = 1.0 l/kg, Cl1 (systemic clearance) = 0.013 l x kg x min, Cl2 = 0.030 l x kg x min. The intersubject variabilities for V1, V2, and Cl1 were 45%, 38%, and 22%, respectively. Plasma concentrations in South African children were 29% less than in Canadian children. The volumes and clearances in the South African children were 29% larger. The terminal half-life was 110 min (1.8 h). Median absolute prediction error for the two-compartment mammillary model was 18%. Heart rate and systolic blood pressure decreased with time and with increasing doses of dexmedetomidine. Respiratory rate and oxygen saturation (in air) were maintained. Sedation was transient. CONCLUSION: The pharmacokinetics of dexmedetomidine in children are predictable with a terminal half-life of 1.8 h. Hemodynamic responses decreased with increasing doses of dexmedetomidine. Respiratory responses were maintained, whereas sedation was transient. PMID- 17122573 TI - Forehead pulse oximetry: Headband use helps alleviate false low readings likely related to venous pulsation artifact. AB - BACKGROUND: This study investigated whether a tensioning headband that applies up to 20 mmHg pressure over a forehead pulse oximetry sensor could improve arterial hemoglobin oxygen saturation reading accuracy in presence of venous pooling and pulsations at the forehead site. METHODS: Healthy volunteers were studied breathing room air in supine and various levels of negative incline (Trendelenburg position) using the forehead sensor with the headband adjusted to its maximum and minimum recommended pressure limits. Saturation readings obtained from the forehead sensor with the subjects supine and the headband in place were used as a baseline to compare the effects of negative incline on reading accuracy when using and not using the headband. Occurrences of false low-saturation readings detected by forehead sensors were compared with those from digit sensors. RESULTS: No difference was observed between saturation readings obtained from the forehead sensor in supine and negative incline positions when the headband was applied. Forehead sensor readings obtained while subjects were inclined and the headband was not used were significantly lower (P < 0.05) than the supine readings. There was no statistically significant difference between the digit and forehead sensor in reporting false low-saturation readings when the headband was applied, regardless of body incline. CONCLUSIONS: Application of up to 20 mmHg pressure on the forehead pulse oximetry sensor using an elastic tensioning headband significantly reduced reading errors and provided consistent performance when subjects were placed between supine and up to 15 degrees head down incline (Trendelenburg position). PMID- 17122574 TI - Estimation of mean body temperature from mean skin and core temperature. AB - BACKGROUND: Mean body temperature (MBT) is the mass-weighted average temperature of body tissues. Core temperature is easy to measure, but direct measurement of peripheral tissue temperature is painful and risky and requires complex calculations. Alternatively MBT can be estimated from core and mean skin temperatures with a formula proposed by Burton in 1935: MBT = 0.64 x TCore + 0.36 x TSkin. This formula remains widely used, but has not been validated in the perioperative period and seems unlikely to remain accurate in dynamic perioperative conditions such as cardiopulmonary bypass. Therefore, the authors tested the hypothesis that MBT, as estimated with Burton's formula, poorly estimates measured MBT at a temperature range between 18 degrees and 36.5 degrees C. METHODS: The authors reevaluated four of their previously published studies in which core and mass-weighted mean peripheral tissue temperatures were measured in patients undergoing substantial thermal perturbations. Peripheral compartment temperatures were estimated using fourth-order regression and integration over volume from 18 intramuscular needle thermocouples, 9 skin temperatures, and "deep" hand and foot temperature. MBT was determined from mass-weighted average of core and peripheral tissue temperatures and estimated from core temperature and mean skin temperature (15 area-weighted sites) using Burton's formula. RESULTS: Nine hundred thirteen data pairs from 44 study subjects were included in the analysis. Measured MBT ranged from 18 degrees to 36.5 degrees C. There was a remarkably good relation between measured and estimated MBT: MBTmeasured = 0.94 x MBTestimated + 2.15, r = 0.98. Differences between the estimated and measured values averaged -0.09 degrees +/- 0.42 degrees C. CONCLUSIONS: The authors concluded that estimation of MBT from mean skin and core temperatures is generally accurate and precise. PMID- 17122575 TI - The effects of ketamine and rocuronium on the A-Line auditory evoked potential index, Bispectral Index, and spectral entropy monitor during steady state propofol and remifentanil anesthesia. AB - BACKGROUND: The authors studied the effects of ketamine and rocuronium on the Bispectral Index, A-Line auditory evoked potential index, state entropy, and response entropy during a calculated steady state anesthesia with propofol and remifentanil. METHODS: After ethics committee approval, 42 patients were allocated to four groups. Baseline measurements were performed after implementing a calculated steady state anesthesia with propofol and remifentanil. The control group received no additional medication. The ketamine group received a bolus and continuous infusion of ketamine. The rocuronium group received a bolus of rocuronium. The rocuronium-ketamine group received both. All data were stored during 15 min after baseline. After inspection of the raw data, the authors conducted an explorative statistical analysis. RESULTS: No significant changes were found in the control group for any of the monitors. Mean values decreased in the rocuronium group for the A-Line auditory evoked potential index, Bispectral Index, and response entropy, but not for state entropy. In the ketamine group, the A-Line auditory evoked potential index and Bispectral Index did not change significantly, but state and response entropy increased. In the rocuronium ketamine group, the A-Line auditory evoked potential index and Bispectral Index did not decrease as found in the rocuronium group. Response and state entropy increased significantly. CONCLUSIONS: The response of all monitors after ketamine administration is not affected by simultaneous administration of rocuronium. Interpretation of all studied indices must be done cautiously while taking into account the clinical setting during measurement. PMID- 17122576 TI - Pharmacokinetics and pharmacodynamics of midazolam and metabolites in nonventilated infants after craniofacial surgery. AB - BACKGROUND: Because information on the optimal dose of midazolam for sedation of nonventilated infants after major surgery is scant, a population pharmacokinetic and pharmacodynamic model is developed for this specific group. METHODS: Twenty four of the 53 evaluated infants (aged 3-24 months) admitted to the Pediatric Surgery Intensive Care Unit, who required sedation judged necessary on the basis of the COMFORT-Behavior score and were randomly assigned to receive midazolam, were included in the analysis. Bispectral Index values were recorded concordantly. Population pharmacokinetic and pharmacodynamic modeling was performed using NONMEM V (GloboMax LLC, Hanover, MD). RESULTS: For midazolam, total clearance was 0.157 l/min, central volume was 3.8 l, peripheral volume was 30.2 l, and intercompartmental clearance was 0.30 l/min. Assuming 60% conversion of midazolam to 1-OH-midazolam, the volume of distribution for 1-OH-midazolam and 1-OH-midazolamglucuronide was 6.7 and 1.7 l, and clearance was 0.21 and 0.047 l/min, respectively. Depth of sedation using COMFORT-Behavior could adequately be described by a baseline, postanesthesia effect (Emax model) and midazolam effect (Emax model).The midazolam concentration at half maximum effect was 0.58 mum with a high interindividual variability of 89%. Using the Bispectral Index, in 57% of the infants the effect of midazolam could not be characterized. CONCLUSION: In nonventilated infants after major surgery, midazolam clearance is two to five times higher than in ventilated children. From the model presented, the recommended initial dosage is a loading dose of 1 mg followed by a continuous infusion of 0.5 mg/h during the night for a COMFORT-Behavior of 12-14 in infants aged 1 yr. Large interindividual variability warrants individual titration of midazolam in these children. PMID- 17122577 TI - Extent and localization of changes in upper airway caliber with varying concentrations of sevoflurane in children. AB - BACKGROUND: Previous studies in humans suggest that inhibition of upper airway muscle activity is independent of the dose of inhalational anesthesia. Whether a dose-independent relation applies to changes in airway caliber is unknown. The authors sought to evaluate the configurational changes that lead to upper airway narrowing during inhalational anesthesia with sevoflurane and to determine whether these changes are dose dependent within a clinically relevant dose range. METHODS: Fifteen children undergoing elective magnetic resonance imaging of the brain were studied. Magnetic resonance images of the upper airway were acquired at sevoflurane concentrations of 0.5, 1.0, and 1.5 minimum alveolar concentration (MAC), administered in random sequence. At least 15 min was allowed for equilibration of inspired and alveolar partial pressures of sevoflurane. Images were acquired in early expiration at the level of the soft palate, base of the tongue, and tip of the epiglottis. Airway cross-sectional area (CSA), anteroposterior, and transverse dimension were determined using image-analysis software. RESULTS: At each anatomical level, pharyngeal CSA decreased progressively with increasing depth of sevoflurane anesthesia (P < 0.001). Increasing the sevoflurane concentration from 0.5 to 1.0 MAC reduced airway CSA by 13-18%, and a further increase to 1.5 MAC resulted in an overall 28-34% reduction in CSA. The reduction in CSA was predominantly due to a decrease in anteroposterior dimension. CONCLUSIONS: Increasing the depth of sevoflurane anesthesia resulted in a relatively uniform reduction in pharyngeal caliber at each anatomical level studied. The effect of sevoflurane on upper airway caliber is dose dependent. PMID- 17122578 TI - A multicenter clinical study of the safety and activity of maleimide-polyethylene glycol-modified Hemoglobin (Hemospan) in patients undergoing major orthopedic surgery. AB - BACKGROUND: Hemospan (Sangart Inc., San Diego, CA), a polyethylene glycol modified hemoglobin with unique oxygen transport properties, has successfully completed a phase I trial in healthy volunteers. Because adverse events are expected to increase with age, the authors conducted a phase II safety study of Hemospan in elderly patients undergoing elective hip arthroplasty during spinal anesthesia. METHODS: Ninety male and female patients, American Society of Anesthesiologists physical status I-III, aged 50-89 yr, in six Swedish academic hospitals were randomly assigned to receive either 250 or 500 ml Hemospan or Ringer's acetate (30 patients/group) before induction of spinal anesthesia. Safety assessment included vital signs and Holter monitoring from infusion to 24 h, evaluation of laboratory values, and fluid balance. The hypothesis to be tested was that the incidence of adverse events would be no more frequent in patients who received Hemospan compared with standard of care (Ringer's acetate). RESULTS: Three serious adverse events were noted, none of which was deemed related to study treatment. Liver enzymes, amylase, and lipase increased transiently in patients in all three groups. There were no significant differences in electrocardiogram or Holter parameters, but there was a suggestion of more bradycardic events in the treated groups. Hypotension was less frequent in the treated patients compared with controls. CONCLUSIONS: In comparison with Ringer's acetate, Hemospan mildly elevates hepatic enzymes and lipase and is associated with less hypotension and more bradycardic events. The absence of a high frequency of serious adverse events suggests that further clinical trials should be undertaken. PMID- 17122579 TI - Pharmacologic and functional characterization of malignant hyperthermia in the R163C RyR1 knock-in mouse. AB - BACKGROUND: Malignant hyperthermia is a pharmacogenetic disorder affecting humans, dogs, pigs, and horses. In the majority of human cases and all cases in animals, malignant hyperthermia has been associated with missense mutations in the skeletal ryanodine receptor (RyR1). METHODS: The authors used a "knock-in" targeting vector to create mice carrying the RyR1 R163C malignant hyperthermia mutation. RESULTS: Validation of this new mouse model of human malignant hyperthermia susceptibility includes (1) proof of transcription of the R163C allele and expression of ryanodine receptor protein in R163C heterozygous and R163C homozygous animals; (2) fulminant malignant hyperthermia episodes in R163C heterozygous mice after exposure to 1.25-1.75% halothane or an ambient temperature of 42 degrees C characterized by increased rectal temperature, respiratory rate, and inspiratory effort, with significant blood biochemical changes indicating metabolic acidosis, ending in death and hyperacute rigor mortis; (3) intraperitoneal pretreatment with dantrolene provided 100% protection from the halothane-triggered fulminant malignant hyperthermia episode; (4) significantly increased sensitivity (decreased effective concentration causing 50% of the maximal response) of R163C heterozygous and homozygous myotubes to caffeine, 4-chloro-m-cresol, and K-induced depolarization; (5) R163C heterozygous and homozygous myotubes have a significantly increased resting intracellular Ca concentration compared with wild type; (6) R163C heterozygous sarcoplasmic reticulum membranes have a twofold higher affinity (Kd = 35.4 nm) for [H]ryanodine binding compared with wild type (Kd = 80.1 nm) and a diminished inhibitory regulation by Mg. CONCLUSIONS: Heterozygous R163C mice represent a valid model for studying the mechanisms that cause the human malignant hyperthermia syndrome. PMID- 17122580 TI - Influence of mannitol and furosemide, alone and in combination, on brain water content after fluid percussion injury. AB - BACKGROUND: Furosemide and mannitol are used to reduce intracranial pressure, but the impact of furosemide on edema of injured brain is unclear. The authors examined the effects of furosemide and mannitol, alone and in combination, on brain water content in brain-injured rats. METHODS: Anesthetized rats were subjected to a 2.2-atm left hemispheric fluid percussion injury. Two and three quarters hours later, animals received 0.5, 1, 4, or 8 g/kg mannitol; 8 mg/kg furosemide; a combination of 4 g/kg mannitol plus 4 mg/kg furosemide; or 8 g/kg mannitol plus 8 mg/kg furosemide. One hour later (4 h after injury), plasma osmolality was measured, and hemispheric water content was determined by drying. Other animals were subjected to injury without drug treatment (impact only) or did not undergo injury (control). Pairwise group comparisons regarding the effects of mannitol and furosemide were restricted to only four groups: impact only, 8 g/kg mannitol, 8 mg/kg furosemide, and 8 g/kg mannitol plus 8 mg/kg furosemide. RESULTS: The water content of both hemispheres in the impact-only group was greater than in the control group (left greater than right). Mannitol, 8 g/kg, increased osmolality from 306 +/- 4 to 351 +/- 6 mOsm/kg (mean +/- SD) and reduced water content in the left hemisphere from 80.06 +/- 0.84% (impact only) to 78.24 +/- 0.73%. Furosemide, 8 mg/kg, had no effect on osmolality or water content. Brain water in animals treated with 8 g/kg mannitol plus 8 mg/kg furosemide did not differ from that seen with 8 g/kg mannitol alone. CONCLUSIONS: Mannitol increased plasma osmolality and reduced water content of the injured and contralateral hemispheres, whereas the authors observed no effect of furosemide when given either alone or in combination with mannitol. PMID- 17122581 TI - Inhibition of glutamatergic activation of extracellular signal-regulated protein kinases in hippocampal neurons by the intravenous anesthetic propofol. AB - BACKGROUND: Intravenous anesthetics cause amnesia, but the underlying molecular mechanisms are poorly understood. Recent studies reveal a significant role of extracellular signal-regulated protein kinases (ERKs) in controlling synaptic plasticity and memory formation. As a major synapse-to-nucleus superhighway, ERK transmits N-methyl-D-aspartate (NMDA) receptor signals to inducible transcriptional events essential for NMDA receptor-dependent forms of synaptic plasticity and memory. This study investigated the role of the widely used intravenous anesthetic propofol in regulating NMDA receptor-dependent ERK phosphorylation. METHODS: The possible effect of propofol on NMDA receptor mediated ERK phosphorylation was detected in cultured rat hippocampal neurons with Western blot analysis. RESULTS: The authors found that propofol at clinical relevant concentrations (1-10 microm) reduced NMDA receptor-mediated ERK phosphorylation. This reduction was independent of gamma-aminobutyric acid transmission. The inhibition of the NMDA receptor seems to contribute to the effect of propofol on NMDA-stimulated ERK phosphorylation, because propofol reduced constitutive NMDA receptor NR1 subunit phosphorylation and impaired NMDA receptor-mediated Ca influx. Furthermore, by inhibiting the ERK pathway, propofol blocked NMDA receptor-dependent activation of two key transcription factors, Elk 1 and cyclic adenosine monophosphate response element-binding protein (CREB), and, as a result, attenuated Elk-1/CREB-dependent reporter gene (c-Fos) expression. CONCLUSIONS: These results suggest that propofol possesses the ability to inhibit NMDA receptor activation of the ERK pathway and subsequent transcriptional activities in hippocampal neurons. These findings indicate a new avenue to explore a transcription-dependent mechanism that may underlie anesthetic interference with synaptic plasticity related to amnesic properties of intravenous anesthetics. PMID- 17122582 TI - Pulmonary responses to overventilation in late multiple organ failure. AB - BACKGROUND: Patients with multiple organ failure (MOF) require mechanical ventilation for several days. The enormous significance of the ventilation strategy for the outcome of these patients is well appreciated. However, most studies have focused on the onset and the early phase of MOF. It was the aim of the current study to investigate the effect of ventilation in the course of MOF. METHODS: Using a model where mice develop MOF 7-14 days after intraperitoneal injection of zymosan, the authors analyzed lung functions, signaling pathways, and mediator release in response to protective ventilation (end-expiratory pressure -3 cm H2O; end-inspiratory pressure -10 cm H2O) and overventilation ( 22.5 cm H2O) in isolated lungs ex vivo. RESULTS: On day 7, pulmonary compliance, pulmonary resistance, and tidal volume were normal, but vascular resistance was elevated compared with untreated animals. During ex vivo ventilation, these lungs showed enhanced nuclear factor-kappaB activation, Akt kinase phosphorylation, and cytokine release, and this was further aggravated by overventilation. After 14 days, zymosan-treated animals were characterized by pulmonary hypertension, reduced tidal volume, elevated pulmonary resistance, and increased mediator production. However, in these lungs, neither nuclear factor-kappaB activation nor cytokine production where enhanced by overventilation. CONCLUSIONS: The zymosan model is characterized by pulmonary inflammation, diminished lung functions, and chronic hypertension. Mechanical ventilation with high distending pressures further augmented cytokine production in this chronic model of MOF only if it significantly augmented tidal volume. The authors speculate that these findings may be explained on the basis of different degrees of lung stretch. PMID- 17122583 TI - Heme oxygenase 1, nuclear factor E2-related factor 2, and nuclear factor kappaB are involved in hemin inhibition of type 2 cationic amino acid transporter expression and L-Arginine transport in stimulated macrophages. AB - BACKGROUND: L-Arginine transport mediated by type 2 cationic amino acid transporter (CAT-2) is one crucial mechanism that regulates nitric oxide production mediated by inducible nitric oxide synthase. Heme oxygenase (HO)-1 induction has been reported to significantly attenuate inducible nitric oxide synthase expression and nitric oxide production. The authors sought to explore the effects of HO-1 induction on CAT-2 expression and L-arginine transport. The effects of HO-1 induction on nuclear factor E2-related factor 2 (Nrf2) and nuclear factor kappaB (NF-kappaB) were also investigated. METHODS: Murine macrophages (RAW264.7 cells) were randomized to receive lipopolysaccharide, lipopolysaccharide plus hemin (an HO-1 inducer; 5, 50, or 500 microm), lipopolysaccharide plus hemin (5, 50, or 500 microm) plus tin protoporphyrin (an HO-1 inhibitor), or lipopolysaccharide plus hemin (5, 50, or 500 microm) plus hemoglobin (a carbon monoxide scavenger). Then, cell cultures were harvested and analyzed. RESULTS: Lipopolysaccharide significantly induced Nrf2 activation and HO-1 expression. Lipopolysaccharide also significantly induced NF-kappaB activation, CAT-2 expression, and L-arginine transport. In a dose-dependent manner, hemin enhanced the lipopolysaccharide-induced Nrf2 activation and HO-1 expression. In contrast, hemin, also in a dose-dependent manner, significantly attenuated the lipopolysaccharide-induced NF-kappaB activation, CAT-2 expression, and L-arginine transport. Furthermore, the effects of hemin were significantly reversed by both tin protoporphyrin and hemoglobin. CONCLUSIONS: HO-1 induction significantly inhibited CAT-2 expression and L-arginine transport in lipopolysaccharide-stimulated macrophages, possibly through mechanisms involved activation of Nrf2 and inhibition of NF-kappaB. In addition, carbon monoxide mediated, at least in part, the effects of HO-1 induction on CAT-2 expression and L-arginine transport. PMID- 17122584 TI - Up-regulation of hypoxia inducible factor 1alpha by isoflurane in Hep3B cells. AB - BACKGROUND: The volatile anesthetic isoflurane induces hypoxia inducible factor (HIF)-1-responsive genes heme oxygenase 1, inducible nitric oxide synthase, and vascular endothelial growth factor (VEGF) expression. Little is known about the extent to which induction of HIF-1alpha is affected by isoflurane. METHODS: Hep3B cells were exposed to isoflurane at various concentrations (0.5-4%) or for different time periods (2-8 h) at 37 degrees C. HIF-1alpha gene expression and transcriptional activity, heme oxygenase 1, inducible nitric oxide synthase, and VEGF gene expression were quantified. RESULTS: Isoflurane induced a time- and concentration-dependent increase in HIF-1alpha protein but not for HIF-1alpha messenger RNA (mRNA) in Hep3B cells. The maximal increase was induced by 2% isoflurane, and the cells incubated with 2% isoflurane for 4-8 h expressed the highest protein. Similarly, HIF-1alpha transcriptional activity was higher in Hep3B cells exposed to 2% isoflurane for 16 h than that in control cells. The combination of 2% isoflurane and desferrioxamine, a hypoxia mimetic, caused a higher level of HIF-1alpha protein than that induced by 2% isoflurane alone. Reoxygenation and inhibitor of proteasome pathway MG132 did not affect the isoflurane-induced HIF-1alpha protein accumulation. Cycloheximide, an inhibitor for protein synthesis, completely abrogated the induction of HIF-1alpha protein by isoflurane. Isoflurane stimulated heme oxygenase 1, inducible nitric oxide synthase, and VEGF mRNA expression in a concentration-dependent manner, and inactivation of HIF-1alpha attenuated the induction of VEGF mRNA by isoflurane. CONCLUSION: Isoflurane can up-regulate HIF-1alpha and enhance HIF-1-responsive genes heme oxygenase 1, inducible nitric oxide synthase, and VEGF mRNA expression in Hep3B cells. The induction of HIF-1alpha by isoflurane does not involve protein degradation but depends on translation pathway. PMID- 17122585 TI - Influence of endothelial glycocalyx degradation and surfactants on air embolism adhesion. AB - BACKGROUND: Microbubble adherence to endothelial cells is enhanced after damage to the glycocalyx. The authors tested the hypothesis that exogenous surfactants delivered intravascularly have differential effects on the rate of restoration of blood flow after heparinase-induced degradation of the endothelial glycocalyx. METHODS: Air microbubbles were injected into the rat cremaster microcirculation after perfusion with heparinase or saline and intravascular administration of either saline or one of two surfactants. The surfactants were Pluronic F-127 (Molecular Probes, Eugene, OR) and Perftoran (OJSC SPC Perftoran, Moscow, Russia). Embolism dimensions and dynamics were observed using intravital microscopy. RESULTS: Significant results were that bubbles embolized the largest diameter vessels after glycocalyx degradation. Bubbles embolized smaller vessels in the surfactant treatment groups. The incidence of bubble dislodgement and the magnitude of distal displacement were smallest after glycocalyx degradation alone and largest after surfactant alone. The time to bubble clearance and restoration of blood flow was longest with heparinase alone and shortest with Pluronic F-127 alone. CONCLUSIONS: Degradation of the glycocalyx causes air bubbles to adhere to the endothelium more proximally in the arteriolar microcirculation. Surfactants added after glycocalyx degradation and before gas embolization promotes bubble lodging in the distal microcirculation. Surfactants may have a clinical role in reducing embolism bubble adhesion to endothelial cells undergoing glycocalyx disruption. PMID- 17122586 TI - Effect of high- and low-molecular-weight low-substituted hydroxyethyl starch on blood coagulation during acute normovolemic hemodilution in pigs. AB - BACKGROUND: Hydroxyethyl starches (HES) with lower impact on blood coagulation but longer intravascular persistence are of clinical interest. The current study aimed to investigate in vivo the isolated effect of molecular weight on blood coagulation during progressive acute normovolemic hemodilution. METHODS: Twenty four pigs were normovolemically hemodiluted up to a total exchange of 50 ml . kg . body weight of HES 650/0.42 or HES 130/0.42. Serial blood sampling was performed to measure HES plasma concentration and to assess blood coagulation. Concentration-effect relations were analyzed by linear regression, followed by the Student t test on regression parameters. RESULTS: Blood coagulation was increasingly compromised toward hypocoagulability by acute normovolemic hemodilution with both treatments (P < 0.01). Significantly greater impact on activated partial thromboplastin time (P = 0.04) and significantly stronger decrease of maximal amplitude (P = 0.04), angle alpha (P = 0.02), and coagulation index (P = 0.02) was seen after acute normovolemic hemodilution with HES 650/0.42 as compared with HES 130/0.42. Except for factor VIII (P = 0.04), no significant differences between both treatments were observed when relating antihemostatic effects to HES plasma concentrations (P > 0.05). A significantly lesser decrease of hemoglobin concentration has been found with HES 650/0.42 as compared with HES 130/0.42 (P < 0.01) in relation to HES plasma concentrations. CONCLUSION: High molecular-weight HES (650/0.42) shows a moderately greater antihemostatic effect than low-molecular-weight HES (130/0.42) during acute normovolemic hemodilution. However, similar effects on hemostasis were observed with both treatments when observed antihemostatic effects were related to measured HES plasma concentrations. In addition, HES 650/0.42 may have a lower efficacy in immediately restoring plasma volume. PMID- 17122587 TI - Retropharyngeal hematoma after stellate ganglion block: Analysis of 27 patients reported in the literature. AB - BACKGROUND: Retropharyngeal hematoma (RPH) is rare; however, it causes airway obstruction and can be fatal. Stellate ganglion block (SGB) can cause RPH. The authors analyzed reports of patients with RPH after SGB to clarify the initial symptoms and signs, and the urgency of airway management. METHODS: MEDLINE and Japana Centra Revuo Medicina were searched for reports of RPH after SGB using the following terms and key words: stellate ganglion block, complication, hematoma, and retropharyngeal hematoma. RESULTS: The authors found 27 patients with RPH after SGB in the past 40 yr. The initial symptoms included neck pain (n = 10), dyspnea (n = 10), neck swelling (n = 8), and hoarseness (n = 5). The symptoms occurred 2 h or more after SGB in 14 patients (52%). Emergency airway management was needed in 21 patients (78%) because of airway obstruction. Among the 21 patients, orotracheal intubation was attempted first in 17 patients; however, it was unsuccessful in 5 patients who immediately needed emergency tracheostomy. Tracheal intubation was impossible by distortion of the anatomy of the markedly edematous pharyngolarynx. Failed airway management caused one death. There were no statistically significant predictors of the initial symptoms or signs for later emergency airway management. CONCLUSIONS: RPH after SGB necessitates emergency airway management. Because airway obstruction cannot be predicted by the initial symptoms or signs, emergency airway management tools should be at hand, and the patency of the airway should be continuously evaluated after onset of RPH after SGB. PMID- 17122588 TI - Strain and sex influence on pain sensitivity after plantar incision in the mouse. AB - BACKGROUND: A small surgical incision in mouse glabrous hind-paw skin induces short-lasting guarding behavior and mechanical and heat hyperalgesia-like behaviors, which imitate human postoperative pain. The increasing popularity of this animal model in drug discovery necessitates the understanding of genetic and sex influence on this animal model. METHODS: The authors examined pain behaviors on DBA2, C57Bl/6, and 129X1/SvJ mice and male and female DBA2 mice before and after plantar incision. RESULTS: The baseline nociceptive responses of these strains were similar, with a few exceptions. Heat responses were different between DBA2 and C57Bl/6 mice, and responses to one filament, 14.0 mN, were intermittently different. Sex did not greatly influence baseline responses. After plantar incision, these three strains of mice were not different in the development of guarding behaviors. Heat responses were only different on postincision day 3 (129X1/SvJ vs. C57Bl/6 mice); otherwise, they were the same. The responses to the series of von Frey filaments were the same after incision in the three strains. Sex did not influence incision-induced pain behaviors in DBA2 mice. CONCLUSION: Although several studies postulated that mouse strain influences pain models, the authors' data indicate that such influence on incisional pain is negligible. This suggests that studies using an incision and knockout mice resulting from 129 strain mutation in a C57BL/6 strain background should have modest influence. The lack of sex differences in incisional pain may encourage researchers to use both male and female subjects in their studies. PMID- 17122589 TI - Value of preoperative clinic visits in identifying issues with potential impact on operating room efficiency. AB - BACKGROUND: Preoperative clinics have been shown to decrease operating room delays and cancellations. One mechanism for this positive economic impact is that medical issues are appropriately identified and necessary information is obtained, so that knowledge of the patients' status is complete before the day of surgery. In this study, the authors describe the identification and management of medical issues in the preoperative clinic. METHODS: All patients coming to the Preoperative Clinic during a 3-month period from November 1, 2003, through January 31, 2004, at the Brigham and Women's Hospital, Boston, Massachusetts, were studied. Data were collected as to the type of issue, information needed to resolve the issue, time to retrieve the information, cancellation and delay rates, and the effect on management. RESULTS: A total of 5,083 patients were seen in the preoperative clinic over the three-month period. A total of 647 patients had a total of 680 medical issues requiring further information or management. Of these issues, 565 were thought to require further information regarding known medical problems, and 115 were new medical problems first identified in the clinic. Most of the new problems required that a new test or consultation be done, whereas most of the old problems required retrieval of information existing from outside medical centers. New problems had a far greater probability of delay (10.7%) or cancellation (6.8%) than old problems (0.6% and 1.8%, respectively). CONCLUSIONS: The preoperative evaluation can identify and resolve a number of medical issues that can impact efficient operating room resource use. PMID- 17122590 TI - Improved postoperative outcomes associated with preoperative statin therapy. AB - Statin therapy is well established for prevention of cardiovascular disease. Statins may also reduce postoperative mortality and morbidity via a pleiotropic (non-lipid-lowering) effect. The authors conducted a meta-analysis to determine the influence of statin treatment on adverse postoperative outcomes in patients undergoing cardiac, vascular, or noncardiovascular surgery. Two independent authors abstracted data from 12 retrospective and 3 prospective trials (n = 223,010 patients). A meta-analysis was performed to evaluate the overall effect of preoperative statin therapy on postoperative outcomes. Preoperative statin therapy was associated with 38% and 59% reduction in the risk of mortality after cardiac (1.9% vs. 3.1%; P = 0.0001) and vascular (1.7% vs. 6.1%; P = 0.0001) surgery, respectively. When including noncardiac surgery, a 44% reduction in mortality (2.2% vs. 3.2%; P = 0.0001) was observed. Preoperative statin therapy may reduce postoperative mortality in patients undergoing surgical procedures. However, the statin associated effects on postoperative cardiovascular morbidity are too variable to draw any conclusion. PMID- 17122591 TI - Delayed presentation of gastric perforation after transesophageal echocardiography for cardiac surgery. PMID- 17122592 TI - Antifibrinolytic therapy use to mitigate blood loss during staged complex major spine surgery: Postoperative visual color changes after tranexamic acid administration. PMID- 17122593 TI - Is bilateral cerebral subdural hematoma more frequent after epidural anesthesia than spinal anesthesia? PMID- 17122595 TI - Contrast imaging while breast-feeding. PMID- 17122596 TI - To sleep, perchance to decode? PMID- 17122600 TI - Role of a reduction of cytokine levels in isoflurane-mediated protection from endotoxin-induced lung injury. PMID- 17122601 TI - Hypotension, heart rate variability, and altered autonomic function. PMID- 17122602 TI - What sunrise might reveal. PMID- 17122604 TI - ProSeal laryngeal mask airway size and fiberoptic endotracheal intubation. PMID- 17122608 TI - Surgeons' self-esteem: A change from too high to too low? PMID- 17122609 TI - An aggressive surgical approach leads to long-term survival in patients with pancreatic endocrine tumors. AB - OBJECTIVE: To evaluate the outcome of reoperations in patients with duodenopancreatic neuroendocrine tumors (PETs) in a tertiary referral center. SUMMARY BACKGROUND DATA: The management of reoperations in PETs is still controversial. METHODS: A total of 125 patients with PETs that underwent surgery between 1987 and 2004 at our institution were retrospectively evaluated. The diagnosis of PETs was based on clinical symptoms, biochemical tests, and histopathology. Patients with at least one reoperation were analyzed regarding clinical characteristics, pathology, operations, and long-term follow-up. RESULTS: A total of 33 patients with a median age of 42 years were identified for this study: 13 patients had gastrinomas, 12 patients had nonfunctional islet cell tumors, 6 patients had insulinomas, and 2 patients had vipomas; 24 patients had sporadic NETs, 9 patients had a MEN-1-syndrome; 27 patients had histologically verified malignant tumors; 33 initial operations and 50 reoperations were performed. The initial procedures comprised 27 resections of the primary tumor and 6 explorative laparotomies; 28 of all reoperations were resections of distant metastases, including 15 liver resections; 19 resections of the pancreas or duodenum were performed during reoperations. The overall morbidity and mortality was 45% and 4.8%, respectively. After a median follow-up of 124 months (range, 16 384 months), 27 of 33 patients are still alive, 12 without evidence of disease. All 6 patients with benign tumors are still alive. The 5-, 10-, and actuarial 25 year survival rate for patients with malignant tumors were 81%, 72%, and 36%, respectively. The survival rate was significantly related to the patients age at time of initial operation and better in patients younger than 50 years compared with patients older than 50 years (P = 0.0007), and the presence or development of metastases (none or lymph node metastases versus distant metastases: P = 0.01). CONCLUSION: We show that an aggressive surgical approach leads to long term survival in patients with malignant PETs. Although long-term cure can only be achieved in a proportion of patients with malignant PETs, significant long term palliation can be achieved. PMID- 17122610 TI - Liver transplantation for hereditary hemorrhagic telangiectasia: Report of the European liver transplant registry. AB - BACKGROUND: Hereditary hemorrhagic telangiectasia (HHT) or Rendu-Osler-Weber disease is a rare disease characterized by the presence of arteriovenous malformations. Hepatic involvement can lead to life-threatening conditions. MATERIAL AND METHODS: Forty patients, reported to the European Liver Transplant Registry, were analyzed to define the role of liver transplantation in the treatment of the hepatic disease form. Indications for transplantation were classified according to Garcia-Tsao: cardiac failure (14 patients), biliary necrosis causing hepatic failure (12 patients), severe portal hypertension (5 patients), cardiac failure and biliary necrosis (6 patients), cardiac failure and portal hypertension (2 patients), and cardiac failure associated with biliary necrosis and portal hypertension (1 patient). Eighteen (81%) of 22 patients had pulmonary artery hypertension. Twelve (30%) patients had pretransplant hepatic interventions. Follow-up was complete for all patients with a mean of 69 months (range, 0-230 months). RESULTS: One-, 5- and 10-year actuarial patient and graft survival rates are 82.5%. Six of the 7 pretransplant procedures performed on the hepatic artery were severely complicated. Cardiovascular function documented in 24 patients improved in 18 patients and remained stable in 5 patients; 1 patient died perioperatively of acute heart failure. Twenty-four (60%) patients had post transplant complications, all but one occurring within the first 4 posttransplant months. Seven (17.5%) patients died perioperatively, 6 of them due to bleeding and 1 due to cardiac failure; 1 (2.5%) patient died late due to chronic rejection. There were 2 possible recurrences. Quality of life markedly improved in all 32 surviving patients. CONCLUSION: The results of the largest reported transplant series in the treatment of hepatic-based HHT are excellent. Elimination of hepatobiliary sepsis and reversal of cardiopulmonary changes dramatically improve quality of life of the recipients. LT should be proposed earlier in the course of symptomatic hepatic HHT presenting with life-threatening conditions. Palliative interventions, especially on the hepatic artery, should be avoided in view of their high (infectious) complication rate. PMID- 17122611 TI - Long-term outcome of split liver transplantation using right extended grafts in adulthood: A matched pair analysis. AB - OBJECTIVE: Shortage of suitable organs led to the development of alternative techniques in liver transplantation. Split liver transplantation (SLT) is well established in pediatric patients. SLT is not completely accepted in adult recipients due to potential increased risk of complications. Despite satisfying results of short-term outcome, there is a leak on information of the long-term outcome. Therefore, we compared the outcome after transplantation of the right extended liver lobe with whole liver transplantation (WLT) using a matched pair's analysis. PATIENTS AND METHODS: From the period of January 1993 to February 2005, 70 SLT recipients were matched with 70 WLT recipients of whole livers. Matching criteria were: 1) indication for transplantation, 2) United Network for Organ Sharing (UNOS) status, 3) recipient age, 4) donor age, 5) cold ischemic time, and 6) year of transplantation. The outcome was analyzed retrospectively. RESULTS: Mean follow-up was 36 months. The 2- and 5-year patient survival rates after SLT and WLT were 86.3% and 82.6%, and 78.4% and 75.6%, respectively (log rank, P = 0.2127). Two- and 5-year graft survival rates were 77.3% and 77.3% after SLT and 71.9% and 65.8% after WLT, respectively (log rank, P = 0.3822). The total biliary complication rate was 11.4% in the SLT group versus 10.0% in the WLT group in the short-term course, while it was 8.5% after SLT and 10.0% after WLT in the long term course. We did not observe significant differences between the groups in term of short- and long-term morbidity. CONCLUSION: Transplantation of the right extended lobe deriving from left lateral splitting of deceased donor livers is followed by the same long-term patient and graft survival, which is known from WLT. There were no differences in the complication rates even in long-term outcome implementing that SLT does not put the adult recipient to an increased early and late risk. Transplantation of the extended right liver lobe provides a safe and efficient procedure in adult patients to expand the number of available grafts. PMID- 17122612 TI - Implication of MYH in colorectal polyposis. AB - OBJECTIVE: The aim of this study was to determine the frequency of MYH mutations in one large population of polyposis patients without APC mutation identified. SUMMARY BACKGROUND DATA: Familial adenomatous polyposis (FAP) is the most known inherited colorectal cancer syndrome. In 70% to 80% of polyposis patients, an APC mutation is found. Patients with polyposis but no APC mutation are considered as APC-muted patients and followed as their relatives accordingly. Biallelic mutation of MYH has been found to responsible of colorectal polyposis and cancer in an autosomal recessive pattern of inheritance. METHODS: Between 1978 and 2004, 433 patients were operated for polyposis. A mutation on APC was identified in 322 patients. Among the remaining patients, 44 were identified as possible MYH-muted patients and contacted, and 31 signed informed consent. Clinical data were obtained from the patients' medical notes. Germline mutation of MYH was searched by sequencing the whole gene. To confirm the deleterious effects of biallelic MYH mutation, transversions on K-ras and APC were searched. RESULTS: There were 9 women and 22 men with a mean age of 53.9 years (range, 22-68 years) at the time of diagnosis. The mean number of polyps was 62.8 (range, 11-266). Eighteen patients (58.1%) had a colorectal cancer. We found biallelic MYH mutation in 6 patients (19.3%; 95% confidence interval, 5.2%-33.5%) and 5 (83.3%) had transversions in K-ras and/or APC. CONCLUSION: MYH is a new gene responsible for about 1.4% of all adenomatous polyposis and about 20% of adenomatous polyposis without APC mutation identified. Search for MYH biallelic mutation in these patients should be systematic as it changes their and relatives'surveillance. PMID- 17122613 TI - Acute nonspecific abdominal pain: A randomized, controlled trial comparing early laparoscopy versus clinical observation. AB - AIMS: To evaluate, in a prospective, randomized, single-institution trial, the role of early laparoscopy in the management of nonspecific abdominal pain (NSAP) in young women. PATIENTS AND METHODS: Women aging from 13 to 45 years, admitted for NSAP at the emergency department, were included in the study. Exclusion criteria were pregnancy, previous appendectomy, contraindications to laparoscopy, diagnosis of malignancy, or chronic disease. NSAP was defined as an abdominal pain in right iliac or hypogastric area lasting more than 6 hours and less than 8 days, without fever, leukocytosis, or obvious peritoneal signs and uncertain diagnosis after physical examination and baseline investigations including abdominal sonography. Patients were randomly assigned to early (<12 hours from admission) laparoscopy group (LAP) or to clinical observation group (OBS). After discharge a follow-up was carried out. RESULTS: From January 2001 to February 2004, 508 female patients without previous abdominal surgery were evaluated in admitting area for acute right iliac or hypogastric abdominal pain, in 373 patients diagnosis was established for obvious signs or with baseline investigations. Of the remaining 135 patients, 31 were excluded from study for various reasons, 53 patients were randomly assigned to LAP and 51 to OBS. Groups were similar for age, mean BMI, white blood cell count, and duration of pain. During hospitalization diagnosis was established in 83.4% of the LAP and in 45.1% of OBS (P < 0.05). Twenty patients of OBS (39.2%) were operated during observation because of worsening of symptoms or appearance of peritoneal sign. Diagnoses in LAP were appendicitis in 16 patients (30.1%), pelvic inflammatory disease in 7 (13.2%), carcinoid in 1 (1.9%), other in 18 (33.9%), no diagnosis in 11 (20.7%); diagnoses in OBS were appendicitis in 3 patients (5.8%), pelvic inflammatory disease in 8 (15.6%), other in 12 (23.5%), and no diagnosis in 28 (54.9%). Mean length of hospital stay was 3.7 +/- 0.8 days in LAP and 4.7 +/- 2.4 days in OBS (P < 0.05); no differences were found regarding mortality, morbidity, radiation dose, and analgesia. Mean follow-up time was 29.3 months (range, 12-60 months) for LAP and 30.6 months for OBS (range, 12-60 months). After 3 months from discharge, 20% of patients in LAP and 52% in OBS had recurrent abdominal pain (P < 0.05); after 12 months, 16% in LAP and 25% in OBS (P = not significant). Six patients in OBS required readmission for surgery. CONCLUSIONS: Compared with active clinical observation, early laparoscopy did not show a clear benefit in women with NSAP. A higher number of diagnosis and a shorter hospital stay in the LAP group did not led to a significant reduction in symptoms recurrences at 1 year. PMID- 17122614 TI - Preoperative endoscopic sphincterotomy versus laparoendoscopic rendezvous in patients with gallbladder and bile duct stones. AB - OBJECTIVE: To compare success rate, length of hospital stay, clinical results, and costs of sequential treatment (endoscopic retrograde cholangiopancreatography followed by laparoscopic cholecystectomy) versus the laparoendoscopic Rendezvous in patients with cholecysto-choledocholithiasis. BACKGROUND: The ideal management of common bile duct (CBD) stones in the era of laparoscopic cholecystectomy (LC) remains controversial. METHODS: A total of 91 elective patients with cholelithiasis and CBD stones diagnosed at magnetic resonance cholangiography (MRC) were included in a prospective, randomized trial. The patients were randomized in 2 groups. Group I patients (45 cases) underwent a preoperative endoscopic retrograde cholangiopancreatography (ERCP) with endoscopic sphincterotomy (ES) followed by LC in the same hospital admission. Group II patients (46 cases) underwent LC associated with intraoperative ERCP and ES according to the rendezvous technique. RESULTS: The rate of CBD clearance was 80% for Group I and 95.6% for Group II (P = 0.06). The morbidity rate was 8.8% in Group I and 6.5% in Group II (P = not significant). No deaths occurred in either group. Hospital stay was shorter in Group II than in Group I: 4.3 days versus 8.0 days (P < 0.0001). There was a significant reduction in mean total cost for group II patients versus group I patients: 2829 euro versus 3834 euro (P < 0.05). CONCLUSIONS: When compared with preoperative ERCP with ES followed by LC, the laparoendoscopic rendezvous technique allows a higher rate of CBD stones clearance, a shorter hospital stay, and a reduction in costs. PMID- 17122615 TI - Is liver resection justified for patients with hepatic metastases from breast cancer? AB - OBJECTIVE: The purpose of this study was to examine our experience with hepatic resection (HR) in a relatively unselected group of patients with breast cancer liver metastases (BCLM). BACKGROUND: Although medical therapies provide limited survival benefit (median survival, 3-15 months), inclusion of HR into the multimodality treatment of patients with BCLM remains controversial. Our approach has been to offer HR to all patients with BCLM, provided that curative hepatic resection was feasible and extrahepatic disease was controlled with medical and/or surgical therapy. METHODS: Outcomes for 85 consecutive patients (all female, median age, 47 years) with BCLM treated with HR from 1984 to 2004 were reviewed. Extrahepatic metastases had been treated prior to HR or were synchronously present in 27 patients (32%). BCLM were solitary in 32 patients (38%) and numbered more than 3 in 26 patients (31%). The prognostic value of each study variable was assessed with log rank tests for univariate analysis and Cox proportional hazard models for multivariate analysis. RESULTS: Within 60 days of major hepatectomy (> or =3 segments, 54 patients) or minor hepatectomy (<3 segments, 31 patients), there was no mortality. The median hospital stay was 9 days with complications occurring in 26% of patients. Microscopically and macroscopically positive margins were present in 18% (R1) and 17% (R2) of patients. Following HR, 28 patients (33%) developed isolated hepatic recurrences, 12 of whom were treated with repeat hepatectomy. At a median follow-up interval of 38 months, 32 patients were alive, yielding median and 5-year overall survivals of 32 months and 37%. Median and 5-year disease-free survivals were 20 months and 21%. Study variables independently associated with poor survival were failure to respond to preoperative chemotherapy (P = 0.008), an R2 resection (P = 0.0001), and the absence of repeat hepatectomy (P = 0.01). CONCLUSIONS: For patients with BCLM, HR is safe and may provide a significant survival benefit over medical therapy alone. Response to preoperative chemotherapy, resection margin, and rehepatectomy for intrahepatic recurrence are key prognostic factors. Importantly, favorable outcomes can be achieved even in patients with medically controlled or surgically resectable extrahepatic disease, indicating that surgery should be considered more frequently in the multidisciplinary care of patients with BCLM. PMID- 17122616 TI - Middle segmental pancreatic resection: An option to treat benign pancreatic body lesions. AB - OBJECTIVE: To clarify whether middle segmental pancreatic resection can be performed with comparable morbidity and mortality to classic pancreatic resections for lesions in the mid-portion of the pancreas. SUMMARY BACKGROUND DATA: Pancreaticoduodenectomies or distal pancreatectomy, traditionally used to treat lesions of the pancreatic body, sacrifice a significant amount of normal pancreatic tissue. Middle segmental pancreatic resection has therefore been introduced to minimize loss of functioning pancreatic tissue. PATIENTS AND METHODS: In a prospective 4-year single-center study, 40 consecutive patients with lesions of the neck or the body of the pancreas underwent a middle segmental pancreatic resection. A matched-pairs analysis comparing middle segmental pancreatic resection with pp-Whipple and distal pancreatectomy was included. RESULTS: Seventeen patients had neoplastic lesions (4 solid malignancies, 9 cystic lesions, 4 neuroendocrine tumors) and 23 patients had focal chronic pancreatitis. Postoperative surgical morbidity was 27.5% and mortality 2.5%. The reoperation rate was 5.0%. Three patients (7.5%) developed pancreatic fistula. Median postoperative hospital stay was 11 days (range, 6-62 days). After a median follow-up of 29 months, 97.4% (38 patients) of the patients were satisfied with the operation. The mean quality of life status (EORTC QLQ-C30) was comparable to a normal control population. Matched-pairs analysis revealed no differences of perioperative parameters (except operation time), morbidity, and mortality. However, endocrine pancreatic function was better preserved (P < 0.05) in patients with middle segmental pancreatic resection. CONCLUSIONS: Middle segmental pancreatic resection is an appropriate procedure for selected patients with tumorous lesions in the mid-portion of the pancreas. It preserves pancreatic parenchyma and function and has a mortality and morbidity rate comparable to other pancreatic resection procedures. PMID- 17122617 TI - A prospective, randomized, controlled trial comparing intermittent portal triad clamping versus ischemic preconditioning with continuous clamping for major liver resection. AB - OBJECTIVE: To evaluate whether ischemic preconditioning (IP) with continuous clamping or intermittent clamping (IC) of the portal triad confers better protection during liver surgery. SUMMARY BACKGROUND DATA: IP and IC are distinct protective approaches against ischemic injury. Since both strategies proved to be superior in randomized controlled trials (RCTs) to continuous inflow occlusion alone, we designed a RCT to compare IP and IC in patients undergoing major liver resection. METHODS: Noncirrhotic patients undergoing major liver resection were randomized to receive IP with inflow occlusion (n = 36) or IC (n = 37). Primary endpoints were postoperative liver injury and intraoperative blood loss. Postoperative liver injury was assessed by peak values of AST (alanine aminotransferase) and ALT (aspartate aminotransferase), as well as the area under the curve (AUC) of the postoperative transaminase course. Secondary endpoints included resection time, the need of blood transfusion, ICU, and hospital stay as well as postoperative complications and mortality. RESULTS: Both groups were comparable regarding demographics, ASA score, type of hepatectomy, duration of inflow occlusion (range, 30-75 minutes), and resection surface. The transection related blood loss was 146 versus 250 mL (P = 0.008), and when standardized to the resection surface 1.2 versus 1.8 mL/cm (P = 0.01) for IP and IC, respectively. Although peak AST, AUCAST, and AUCALT were lower for IC, the differences did not reach statistical significance. Overall (42% vs. 38%) and major (33 vs. 27%) postoperative complications as well as median ICU (1 vs. 1 day) and hospital stay (10 vs. 11 days) were similar between both groups. CONCLUSIONS: Both IP and IC appear to be equally effective in protecting against postoperative liver injury in noncirrhotic patients undergoing major liver resection. However, IP is associated with lower blood loss and shorter transection time. Therefore, both strategies can be recommended for noncirrhotic patients undergoing liver resection. PMID- 17122618 TI - Assessment of complications after pancreatic surgery: A novel grading system applied to 633 patients undergoing pancreaticoduodenectomy. AB - OBJECTIVE: To define a simple and reproducible classification of complications following pancreaticoduodenectomy (PD) based on a therapy-oriented severity grading system. BACKGROUND: While mortality is rare after PD, morbidity rates remain high. The lack of standardization in evaluating morbidity after PD has severely hampered meaningful comparisons over time and among centers. We adapted a novel classification of complication to stratify morbidity by severity after PD, to test whether the incidence of pancreatic fistula has changed over time, and to identify risk factors in a single North American center. METHODS: The classification was applied to a consecutive series of 633 patients undergoing PD between February 2003 and August 2005. Another series of 141 patients treated between 1987 and 1990 was also analyzed to identify changes in the incidence and severity of fistula. Univariate and multivariate analyses were performed to link respective complications with preoperative and intraoperative parameters, length of hospital stay, and long-term survival. RESULTS: A total of 263 (41.5%) patients did not develop any complication, while 370 (58.5%) had at least one complication; 62 (10.0%) patients had only grade I complications (no need for specific intervention), 192 patients (30.0%) had grade II (need for drug therapy such as antibiotics), 85 patients (13.5%) had grade III (need for invasive therapy), and 19 patients (3.0%) had grade IV complications (organ dysfunction with ICU stay). Grade V (death) occurred in 12 patients (2.0%). A total of 57 patients (9.0%) developed pancreatic fistula, of which 33 (58.0%) were classified as grade II, 17 (30.0%) as grade III, 5 (9.0%) as grade IV, and 2 (3.5%) as grade V. Delayed gastric emptying was documented in 80 patients (12.7%); half of them were scored as grade II and the other half as grade III. A significant decrease in the incidence of fistula was observed between the 2 periods analyzed (14.0% vs. 9.0%, P < 0.001), mostly due to a decrease in grade II fistula. Cardiovascular disease was a risk factor for overall morbidity and complication severity, while texture of the gland and cardiovascular disease were risk factors for pancreatic fistula. CONCLUSION: This study demonstrates the applicability and utility of a new classification in grading complications following pancreatic surgery. This novel approach may provide a standardized, objective, and reproducible assessment of pancreas surgery enabling meaningful comparison among centers and over time. PMID- 17122619 TI - Long-term follow-up in small duct chronic pancreatitis: A plea for extended drainage by "V-shaped excision" of the anterior aspect of the pancreas. AB - BACKGROUND: A pancreatic duct diameter (PDD) ranging from 4 to 5 mm is regarded as "normal." The "large duct" form of chronic pancreatitis (CP) with a PDD >7 mm is considered a classic indication for drainage procedures. In contrast, in patients with so-called "small duct chronic pancreatitis" (SDP) with a PDD <3 mm extended resectional procedures and even, in terms of an "ultima ratio," total pancreatectomy are suggested. METHODS: Between 1992 and 2004, a total of 644 patients were operated on for CP. Forty-one prospectively evaluated patients with SDP underwent a new surgical technique aiming at drainage of the entire major PD (longitudinal "V-shaped excision" of the anterior aspect of the pancreas). Preoperative workup for imaging ductal anatomy included ERCP/MRCP, visualizing the PD throughout the entire gland. The interval between symptoms and therapeutic intervention varied from 12 to 120 months. Median follow-up was 83 months (range, 39-117 months). A pain score as well as a multidimensional psychometric quality of-life questionnaire was used. RESULTS: Hospital mortality was 0%. The perioperative (30 days) morbidity was 19.6%. Postoperative, radiologic imaging showed an excellent drainage of the entire gland and the PD in all but 1 patient. Global quality-of-life index increased in median by 54% (range, 37.5%-80%). Median pain score decreased by 95%. Twenty-seven patients (73%) had complete pain relief. Sixteen patients (43%) developed diabetes, while the exocrine pancreatic function was well preserved in 29 patients (78%). CONCLUSION: "V-shaped excision" of the anterior aspect of the pancreas is a secure and effective approach for SDP, achieving significant improvement in quality of life and pain relief, hereby sparing patients from unnecessary, extended resectional procedures. The deterioration of exocrine and endocrine pancreatic functions is comparable with that observed during the natural course of the disease. PMID- 17122620 TI - Advanced leg salvage of the critically ischemic leg with major tissue loss by vascular and plastic surgeon teamwork: Long-term outcome. AB - OBJECTIVE: To assess long-term outcome and prognostic factors for extreme surgery by vascular and plastic surgical teamwork for leg salvage in patients with critically ischemic large tissue defects. SUMMARY BACKGROUND DATA: Combined vascular reconstruction and microvascular free-flap transfer has been used to improve distal perfusion and cover large tissue defects caused by the critical limb ischemia (CLI) in few dedicated centers during the past 15 years. Comorbidities compromise the results of these demanding operations, and it is unclear how far this mode of treatment should be extended. METHODS: During 1989 to 2003, altogether 2157 vascular or endovascular revascularizations for CLI manifested as tissue lesions were performed. These included 81 revascularizations combined with microvascular free flap transfers in 79 patients (37-85 years). All the patients were candidates for major amputation. The patients were followed up at least 2 years or to death (mean follow-up, 62 months; SD, +/-34 months). RESULTS: One- and 5-year leg salvage rates were 73% and 66%, survival rates 91% and 63%, and amputation-free survival rates of 70% and 41%, respectively. Male gender and American Society of Anesthesiologists score 4 were associated with an increased risk of death, whereas the involvement of the heel mostly with calcaneal osteomyelitis and a large size of defect predicted major amputation. CONCLUSIONS: A combined vascular reconstruction and free-flap transfer offers an option for advanced limb salvage in a selected group of patients with CLI and a major tissue defect. Poor general condition, the involvement of the heel, and a large defect would indicate an amputation over extreme attempts for limb salvage. PMID- 17122621 TI - Early nasogastric feeding in predicted severe acute pancreatitis: A clinical, randomized study. AB - OBJECTIVE: To compare the efficacy and safety of early, nasogastric enteral nutrition (EN) with total parenteral nutrition (TPN) in patients with predicted severe acute pancreatitis (SAP). SUMMARY BACKGROUND DATA: In SAP, the magnitude of the inflammatory response as well as increased intestinal permeability correlates with outcome. Enteral feeding has been suggested superior to parenteral feeding due to a proposed beneficial effect on the gut barrier. METHODS: Fifty patients who met the inclusion criteria were randomized to TPN or EN groups. The nutritional regimen was started within 24 hours from admission and EN was provided through a nasogastric tube. The observation period was 10 days. Intestinal permeability was measured by excretion of polyethylene glycol (PEG) and concentrations of antiendotoxin core antibodies (Endocab). Interleukins (IL) 6 IL-8, and C-reactive protein (CRP) were used as markers of the systemic inflammatory response. Morbidity and feasibility of the nutritional route were evaluated by the frequency of complications, gastrointestinal symptoms, and abdominal pain. RESULTS: PEG, Endocab, CRP, IL-6, APACHE II score, severity according to the Atlanta classification (22 patients), and gastrointestinal symptoms or abdominal pain did not significantly differ between the groups. The incidence of hyperglycemia was significantly higher in TPN patients (21 of 26 vs. 7 of 23; P < 0.001). Total complications (25 vs. 52; P = 0.04) and pulmonary complications (10 vs. 21; P = 0.04) were significantly more frequent in EN patients, although complications were diagnosed dominantly within the first 3 days. CONCLUSION: In predicted SAP, nasogastric early EN was feasible and resulted in better control of blood glucose levels, although the overall early complication rate was higher in the EN group. No beneficial effects on intestinal permeability or the inflammatory response were seen by EN treatment. PMID- 17122622 TI - Novel short-term hypothermic oxygenated perfusion (HOPE) system prevents injury in rat liver graft from non-heart beating donor. AB - OBJECTIVE: To assess a machine perfusion system in rescuing liver grafts from non heart-beating donors (NHBD). SUMMARY BACKGROUND DATA: The introduction of extracorporeal liver perfusion systems in the clinical routine depends on feasibility. Conceivably, perfusion could be performed during recipient preparation. We investigated whether a novel rat liver machine perfusion applied after in situ ischemia and cold storage can rescue NHBD liver grafts. METHODS: We induced cardiac arrest in male Brown Norway rats by phrenotomy and ligation of the subcardial aorta. We studied 2 experimental groups: 45 minutes of warm in situ ischemia + 5 hours cold storage versus 45 minutes of warm in situ ischemia + 5 hours cold storage followed by 1 hour hypothermic oxygenated extracorporeal perfusion (HOPE). In both groups, livers were reperfused in a closed sanguineous isolated liver perfusion device for 3 hours at 37 degrees C. To test the benefit of HOPE on survival, we performed orthotopic liver transplantation in both experimental groups. RESULTS: After cold storage and reperfusion, NHBD livers showed necrosis of hepatocytes, increased release of AST, and decreased bile flow. HOPE improved NHBD livers significantly with a reduction of necrosis, less AST release, and increased bile flow. ATP was severely depleted in cold-stored NHBD livers but restored in livers treated by HOPE. After orthotopic liver transplantation, grafts treated by HOPE demonstrated a significant extension on animal survival. CONCLUSIONS: We demonstrate a beneficial effect of HOPE by preventing reperfusion injury in a clinically relevant NHBD model. PMID- 17122623 TI - Precultivation of engineered human nasal cartilage enhances the mechanical properties relevant for use in facial reconstructive surgery. AB - OBJECTIVE: To investigate if precultivation of human engineered nasal cartilage grafts of clinically relevant size would increase the suture retention strength at implantation and the tensile and bending stiffness 2 weeks after implantation. SUMMARY BACKGROUND INFORMATION: To be used for reconstruction of nasal cartilage defects, engineered grafts need to be reliably sutured at implantation and resist to bending/tension forces about 2 weeks after surgery, when fixation is typically removed. METHODS: Nasal septum chondrocytes from 4 donors were expanded for 2 passages and statically loaded on 15 x 5 x 2-mm size nonwoven meshes of esterified hyaluronan (Hyaff-11). Constructs were implanted for 2 weeks in nude mice between muscle fascia and subcutaneous tissue either directly after cell seeding or after 2 or 4 weeks of preculture in chondrogenic medium. Engineered tissues and native nasal cartilage were assessed histologically, biochemically, and biomechanically. RESULTS: Engineered constructs reproducibly developed with culture time into cartilaginous tissues with increasing content of glycosaminoglycans and collagen type II. Suture retention strength was significantly higher (3.6 +/- 2.2-fold) in 2-week precultured constructs than in freshly seeded meshes. Following in vivo implantation, tissues further developed and maintained the original scaffold size and shape. The bending stiffness was significantly higher (1.8 +/- 0.8-fold) if constructs were precultured for 2 weeks than if they were directly implanted, whereas tensile stiffness was close to native cartilage in all groups. CONCLUSION: In our experimental setup, preculture for 2 weeks was necessary to engineer nasal cartilage grafts with enhanced mechanical properties relevant for clinical use in facial reconstructive surgery. PMID- 17122624 TI - Tumor infiltrating T lymphocytes in colorectal cancer: Tumor-selective activation and cytotoxic activity in situ. AB - OBJECTIVE: To examine whether tumor-selective infiltration, activation, and cytotoxic activity of tumor infiltrating T lymphocytes (TIL) can be demonstrated in situ in colorectal cancer samples. SUMMARY BACKGROUND DATA: Recent studies indicated a correlation between the presence of TIL and an improved prognosis in colorectal cancer. However, tumor-selective activation and cytotoxic activity of CD8 TIL in situ in colorectal cancer patients have not yet been examined. METHODS: Tumor samples from 49 patients and corresponding normal mucosa samples from 23 patients with colorectal cancer (UICC stages II-IV) were examined for TIL. Two-color fluorescence immunohistochemistry and multicolor flowcytometric (FACS) analysis were used for quantification of CD8 T cells and measurement of their activation status (CD69-expression) and cytotoxic activity (CD107a expression) in situ. Presence of tumor antigen-reactive T cells in tumor, blood, and bone marrow was evaluated by IFN-gamma Elispot analysis. RESULTS: While absolute numbers of CD8 T cells were similar, CD4 T helper cells were significantly increased in tumor tissue compared with normal mucosa. There was a significantly higher proportion of activated and cytotoxically active CD8 TIL in colorectal cancer compared with normal mucosa. Increased activation, cytotoxic activity, and functional reactivity of TIL were correlated with the presence of functional tumor antigen-reactive T cells in the blood and bone marrow. The proportion of activated TIL decreased significantly with higher tumor stage. CONCLUSIONS: Tumor-selective activation and cytotoxic activity of CD8 TIL and tumor-selective migration of CD4 T helper cells were demonstrated in colorectal cancer for the first time. Our data support the immunogenicity of colorectal cancer and suggest clinical significance of tumor-specific immune responses. PMID- 17122625 TI - Prediction of postoperative complications after urgent laparotomy by intraperitoneal microdialysis: A pilot study. AB - OBJECTIVE: The aim of the present study was to investigate the role of intraperitoneal microdialysis (IPM) techniques in monitoring the evolution of postoperative critically ill patients requiring urgent laparotomy. SUMMARY BACKGROUND DATA: Postoperative intraabdominal sepsis is associated with an important degree of morbidity and mortality in acutely ill patients. Early diagnosis is critical to improve outcomes. METHODS: : The study included 25 consecutive patients admitted to the intensive care unit (ICU) after urgent laparotomy. Measurements of microdialysate fluid were performed through a microdialysis catheter, positioned intraperitoneally, during the first 5 postoperative days and lactate/pyruvate (L/P) ratios calculated. Patients were followed until hospital discharge. RESULTS: Ten patients had a complicated postoperative course, including 4 deaths (3 refractory shock, 1 mesenteric ischemia), 3 reinterventions (1 necrotic collection, 1 mesenteric ischemia, 1 biliary leak), 2 secondary peritonitis, and 1 intraabdominal collection. The IPM L/P ratio in these patients was already significantly higher during the first 24 postoperative hours compared with patients who had no complications (35 +/- 21 vs. 18 +/- 6, P < 0.01). An IPM L/P ratio above 22 on postoperative day 1 had a sensitivity of 0.64 and a specificity of 0.79 for complications. There were no significant differences between the two groups in pH, lactate, white blood cell count, or subcutaneous L/P ratio. No complication was associated with the technique. CONCLUSIONS: IPM is safe and reliable and provides valuable information after urgent laparotomy. Persistently high L/P values should raise the possibility of serious postoperative complications. PMID- 17122626 TI - Relation of surgeon and hospital volume to processes and outcomes of colorectal cancer surgery. AB - BACKGROUND: Greater hospital volume has been associated with lower mortality after colorectal cancer surgery. The contribution of surgeon volume to processes and outcomes of care is less well understood. We assessed the relation of surgeon and hospital volume to postoperative and overall mortality, colostomy rates, and use of adjuvant radiation therapy. METHODS: From the California Cancer Registry, we studied 28,644 patients who underwent surgical resection of stage I to III colorectal cancer during 1996 to 1999 and were followed up to 6 years after surgery to assess 30-day postoperative mortality, overall long-term mortality, permanent colostomy, and use of adjuvant radiation therapy. RESULTS: Across decreasing quartiles of hospital and surgeon volume, 30-day postoperative mortality ranged from 2.7% to 4.2% (P < 0.001). Adjusting for age, stage, comorbidity, and median income among patients with colorectal cancer who survived at least 30 days, patients in the lowest quartile of surgeon volume had a higher adjusted overall mortality rate than those in the highest quartile (hazard ratio, 1.16; 95% confidence interval, 1.09-1.24), as did patients in the lowest quartile of hospital volume relative to those treated in the highest quartile (hazard ratio, 1.11; 95% confidence interval, 1.05-1.19). For rectal cancer, adjusted colostomy rates were significantly higher for low-volume surgeons, and the use of adjuvant radiation therapy was significantly lower for low-volume hospitals. CONCLUSIONS: Greater surgeon and hospital volumes were associated with improved outcomes for patients undergoing surgery for colorectal cancer. Further study of processes that led to these differences may improve the quality of colorectal cancer care. PMID- 17122627 TI - The role of abdominoperineal resection as surgical therapy for anorectal melanoma. AB - OBJECTIVES: 1) Characterize changes in the surgical treatment of anorectal melanoma over time. 2) Determine if the extent of surgical resection is associated with outcome. 3) Identify prognostic factors correlating with survival. SUMMARY BACKGROUND DATA: Although early data suggested improved survival in patients undergoing abdominoperineal resection (APR) for primary anorectal melanoma, such an aggressive approach may be unwarranted as distant relapse rates are high. We have seen a trend toward less aggressive surgical treatment of the local disease over the past 20 years. METHODS: A retrospective review was performed of all patients with anorectal melanoma treated at our institution between 1984 and 2003. Extent of primary resection and pathologic factors were studied. RESULTS: Forty-six patients underwent a curative resection with a median follow-up of 29 months, and 5-year disease-specific survival (DSS) rate of 35%. While patient and tumor characteristics remained similar, there was a dramatic shift in surgical treatment toward less radical procedures. Prior to 1997, the majority of patients (15 of 21, 71%) underwent APR. After 1997, the majority of patients (21 of 25, 84%) underwent local excision (LE) (P < 0.0001). Local recurrence was noted in 11 of 46 (24%) patients: 4 of 19 (21%) who underwent APR and 7 of 27 (26%) who underwent LE (P = not significant). Five-year DSS was similar: 34% following APR and 35% following LE. Tumor perineural invasion (PNI) was the only factor identified as an independent predictor of worse outcome (P = 0.01). CONCLUSION: The extent of surgical treatment is not associated with outcome in primary anorectal melanoma. Therefore, LE of the primary tumor is recommended when technically feasible. The presence of PNI is an important prognostic factor and should be considered in future clinical trials. PMID- 17122628 TI - Suicide bombers form a new injury profile. AB - BACKGROUND: Recent explosions of suicide bombers introduced new and unique profiles of injury. Explosives frequently included small metal parts, increasing severity of injuries, challenging both physicians and healthcare systems. Timely detonation in crowded and confined spaces further increased explosion effect. METHODS: Israel National Trauma Registry data on hospitalized terror casualties between October 1, 2000 and December 31, 2004 were analyzed. RESULTS: A total of 1155 patients injured by explosion were studied. Nearly 30% suffered severe to critical injuries (ISS > or = 16); severe injuries (AIS > or = 3) were more prevalent than in other trauma. Triage has changed as metal parts contained in bombs penetrate the human body with great force and may result in tiny entry wounds easily concealed by hair, clothes etc. A total of 36.6% had a computed tomography (CT), 26.8% had ultrasound scanning, and 53.2% had an x-ray in the emergency department. From the emergency department, 28.3% went directly to the operating room, 10.1% to the intensive care unit, and 58.4% directly to the ward. Injuries were mostly internal, open wounds, and burns, with an excess of injuries to nerves and to blood vessels compared with other trauma mechanisms. A high rate of surgical procedures was recorded, including thoracotomies, laparotomies, craniotomies, and vascular surgery. In certain cases, there were simultaneous multiple injuries that required competing forms of treatment, such as burns and blast lung. CONCLUSIONS: Bombs containing metal fragments detonated by suicide bombers in crowded locations change patterns and severity of injury in a civil population. Specific injuries will require tailored approaches, an open mind, and close collaboration and cooperation between trauma surgeons to share experience, opinions, and ideas. Findings presented have implications for triage, diagnosis, treatment, hospital organization, and the definition of surge capacity. PMID- 17122629 TI - Oncologic outcomes after neoadjuvant chemoradiation followed by curative resection with tumor-specific mesorectal excision for fixed locally advanced rectal cancer: Impact of postirradiated pathologic downstaging on local recurrence and survival. AB - OBJECTIVE: The purpose of this study was to determine the oncologic outcomes and clinical factors affecting survival in patients who underwent neoadjuvant chemoradiotherapy following tumor specific mesorectal excision for locally advanced, fixed rectal cancer. SUMMARY BACKGROUND DATA: Neoadjuvant chemoradiation therapy has resulted in significant tumor downstaging, which enhances curative resection and subsequently improves local disease control for rectal cancer. However, oncologic outcomes, according to clinical factors, have not yet been fully understood in locally advanced and fixed rectal cancer. METHODS: A total of 114 patients who had undergone neoadjuvant chemoradiation for advanced rectal cancer (T3 or T4 and node positive) were investigated retrospectively. Chemotherapy was administered intravenously with 5-FU and leucovorin during weeks 1 and 5 of radiotherapy. The total radiation dose was 5040 cGY in 25 fractions delivered over 5 weeks. Tumor-specific mesorectal excision was done 4 to 6 weeks after the completion of neoadjuvant chemoradiation. Survival and recurrence rates, according to the pathologic stage, were evaluated. Moreover, factors affecting survival were investigated. RESULTS: The 5-year survival rates according to pathologic stage were: 100% in pathologic complete remission (n = 10), 80% in stage I (n = 23), 56.8% in stage II (n = 34), and 42.3% in stage III (n = 47) (P = 0.0000). Local, systemic, and combined recurrence rates were 11.4%, 22.8%, and 3.5%, respectively. Multivariate analysis showed that the pathologic N stage and operation method were the independent factors affecting survival rate. CONCLUSION: Pathologic complete remission showed excellent oncologic outcomes, and the pathologic N stage was the most important factor for oncologic outcomes. PMID- 17122630 TI - Abnormal expression of biomarkers in incompletely ablated Barrett's esophagus. AB - OBJECTIVE: The aim of this study was to evaluate expression of cancer risk associated biomarkers in columnar epithelium at squamocolumnar junctions produced by an ablation procedure and proton pump inhibitors in incompletely ablated Barrett's esophagus (BE) patients that were nondysplastic prior to ablation. SUMMARY BACKGROUND DATA: Ablation of BE to squamous epithelium is achievable by combining a re-injury method with acid suppression. We previously reported that, when there is complete ablation, the neo-squamous epithelium is normal histologically and in biomarker expression. However, squamous islands observed after prolonged use of PPIs were associated with abnormalities in p53 expression and Ki-67 labeling. METHODS: Twenty-one nondysplastic BE cases with incomplete ablation were evaluated for the expression of Ki-67 (proliferation), cyclooxygenase-2 (COX-2), and p53 by immunohistochemistry. RESULTS: Pre-ablation biopsies showed the normal staining patterns in columnar epithelium, ie, normal Ki-67 labeling, rare positive COX-2 staining of interstitial cells, and negative or mild staining for p53 in the majority of patients' biopsies. However, post ablation biopsies demonstrated abnormal staining patterns in the glandular area at the new squamocolumnar junctions. In 13 of 21 post-ablation cases (62%), increased Ki-67 staining was seen in BE glands. In 8 of 21 patients (38%), increased COX-2 expression was seen in columnar epithelium. Similarly, in 8 of 21 post-ablation junctions (38%), there was increased p53 staining. CONCLUSIONS: Our findings of increased expression of cancer-associated biomarkers in incompletely ablated BE patients raise a cautionary note regarding this procedure. We hypothesize that newly formed junctions contain cells undergoing replication, differentiation, etc, and are thus more susceptible to genomic damage. PMID- 17122631 TI - CD56-positive large B-cell lymphoma. AB - CD56 (NCAM), a neural adhesion molecule, is normally expressed on natural killer cells and subsets of T cells and is commonly seen on hematolymphoid neoplasms such as plasma cell myeloma and acute myelogenous leukemia. It is uncommon in B cell lymphoma. From 2001 to 2003 a cohort of 20 cases of CD56 B-cell lymphomas was identified by flow cytometry (<0.5% of all B-cell lymphomas studied) during a 2-year period. Most (90%) expressed CD10 and 5/5 tested cases were BCL6, suggesting a follicular origin. An extranodal disease presentation was seen in 45% and may be related to CD56 expression. These CD56 B-cell lymphomas may represent a new subset of large B-cell lymphoma. The relationship of cells with this antigenic profile to normal B-cell differentiation is explored. PMID- 17122632 TI - An inverse relation between COX-2 and E-cadherin expression correlates with aggressive histologic features in prostate cancer. AB - The identification of biomarkers in prostatic carcinoma has yielded important data regarding prognosis and has aided in increasing diagnostic accuracy. Additionally, this approach has yielded important insights into the biology of prostatic carcinoma. In this study, we report that the expression of the cyclooxygenase isoenzyme, COX-2, is significantly increased in prostatic carcinoma, whereas that of the cell adhesion molecule, E-cadherin, is decreased. The expression of COX-2 was positively correlated with higher tumor stage, and the presence of carcinoma in surgical margins at prostatectomy. Conversely, the expression of E-cadherin was inversely related to these prognostic indicators. Lastly, the expressions of COX-2 and E-cadherin were very strongly and inversely correlated. These results provide important insights into the biologic underpinnings of prostate carcinoma; and further studies into COX-2 expression in prostate core biopsies may show utility in preprostatectomy prognostication. Furthermore, these results may provide a rational basis for therapeutic intervention and chemoprevention with COX-2 inhibitor therapy in prostate carcinoma. PMID- 17122633 TI - Refinement of immunohistologic parameters for Her2/neu scoring validation by FISH and CISH. AB - The conventional method of scoring Her2/neu immunostaining is recognized to result in a high false-positive rate among 2+ cases when compared with results obtained with fluorescence in situ hybridization (FISH); however, costs and convenience dictates that immunohistochemistry remains the screening test for Her2/neu status in patients with breast cancer. We describe refined criteria for scoring of Her2/neu on the basis of anatomic localization rather than the subjective assessment of intensity. The presence of a circumferential tram track pattern that results from the staining of apposing cell membranes in >25% of the tumor cells was necessary for a 3+ score (Her2/neu overexpressed) and the presence of the tram track pattern in <25% was scored 2+ (equivocal); granular and fragmented membrane staining was scored 1+ (negative). The tram track pattern of Her2/neu overexpression showed 100% concordance with gene amplification. FISH and CISH testing in selected cases from the other categories validated the revised scoring method. These criteria reduced the numbers of equivocal staining cases that required FISH testing. PMID- 17122634 TI - Strong PDGFRA positivity is seen in GISTs but not in other intra-abdominal mesenchymal tumors: Immunohistochemical and mutational analyses. AB - Mutation of the platelet-derived growth factor receptor-alpha (PDGFRA) gene has been well documented as an alternative oncogenic mechanism in a subset of gastrointestinal stromal tumors (GISTs) lacking c-kit mutations. However, the role of PDGFRA immunohistochemistry in the diagnosis of GISTs has not been well studied. We investigated PDGFRA immunoreactivity in GISTs and in other intra abdominal mesenchymal tumors, and correlated PDGFRA expression with CD117 positivity and with the mutational status of PDGFRA and c-kit genes. In addition, expression of phosphorylated AKT, an activated downstream molecule in the PDGFRA and c-kit signaling pathways, was correlated with PDGFRA and CD117 status. A total of 39 GISTs and 20 other mesenchymal tumors in the abdomen were included in this study. Thirty-five of 39 GIST cases (89.7%) were positive for PDGFRA and 19 of these 35 positive cases were strongly positive. Five of 20 non-GIST lesions (25%) were positive for PDGFRA, but none of these cases were strongly positive. With one exception, PDGFRA-positive cases were also positive for CD117. Phosphorylated AKT positivity was not associated with the immunoreactivity or mutation of PDGFRA and c-kit, suggesting that the activation of AKT is probably independent of the activation of PDGFRA and c-kit in GISTs. Of 14 GISTs assayed, 4 had mutations in c-kit at exons 11 or 17, and 4 had mutations in PDGFRA at exons 12 or 18. Three of 4 GIST cases with PDGFRA mutations show epithelioid morphology and strong PDGFRA immunoreactivity with prominent perinuclear dotlike accentuation (so-called Golgi pattern). In conclusion, strong PDGFRA positivity with Golgi pattern is a useful adjunct in the diagnosis of GISTs with PDGFRA mutation. PMID- 17122635 TI - Aberrant expression of the Rb pathway proteins in soft tissue sarcomas. AB - Cell cycle regulation depends on a fine balance between cyclins, cyclin-dependent kinases (CDKs), and cyclin-dependent kinase inhibitors (CKIs) that block the cycle progression. Alterations of the cell cycle regulators are a common feature of many malignant tumors, and some have been shown to have prognostic significance. In this study, 152 cases of different types of soft tissue sarcomas were evaluated for alterations of cell cycle regulator proteins that control the cell cycle progression from G1 to S phase and govern the Rb pathway. Immunohistochemical stains for proteins Rb, E2F1, cyclin D1, CDK4, CDK6, p16, and p27 were carried out on tissue microarrays. The relationship between the expression of these proteins and the histologic grade of the sarcomas was assessed. Altered expression for Rb and p16 proteins was identified in 67.8% and 65.1% of the cases, respectively. Overexpression of E2F1, cyclin D1, CDK4, and CDK6 was detected in 50.7%, 24.3%, 92.1%, and 10.5%, respectively. Overexpression of E2F1 was associated with altered expression of Rb protein. Overexpression of cyclin D1, CDK4, and CDK6 showed an association with normal Rb expression. CDK6 expression revealed a positive correlation with the histologic grade of the sarcoma, and p27 expression was inversely correlated with sarcoma grade. These results suggest that alterations of the Rb pathway proteins are common in soft tissue sarcomas and may participate in their tumorigenesis. CDK6 and p27 showed correlation with the histologic grade of the sarcomas, suggesting that these proteins could be used as prognostic markers. PMID- 17122636 TI - Sarcomas often express constitutive nitric oxide synthases (NOS) but infrequently inducible NOS. AB - Nitric oxide (NO) has a dual action in tumors, with both pro-tumor and anti-tumor activities. NO is produced by nitric oxide synthases (NOS). There are three enzyme isoforms: two of them are constitutively produced (neuronal or brain NOS and endothelial NOS), and one is an inducible form (iNOS). NOS expression has been shown in several epithelial tumors, but there is no report addressing NOS expression in sarcomas. The authors evaluated the expression of NOS in 97 cases of various sarcomas spotted in duplicate in a tissue array paraffin block. Eighty four of the 97 tumor specimens (86.6%) expressed nNOS, and most of them showed a strong expression of the isoenzyme. Only chondrosarcomas and liposarcomas had significant numbers of negative cases, and all pleomorphic sarcomas, alveolar soft part sarcomas, angiosarcomas, gastrointestinal stromal tumors, and synovial sarcomas showed some degree of positivity. Forty-three cases (44.4%) showed eNOS immunostaining, but only 15.5% showed a strong signal, with emphasis on angiosarcomas, chondrosarcomas, alveolar soft part sarcomas, and synovial sarcoma. Strong expression of iNOS was observed in only 9 cases (9.3%), with weak expression in another 26 cases (26.8%). Strong expression of iNOS was found in malignant peripheral nerve sheet tumors, liposarcomas, pleomorphic sarcomas, fibrosarcomas, chondrosarcomas, and synovial sarcomas. Apparently alveolar soft part sarcomas are unusual in their capacity of expression of NOS isoforms, and in a very peculiar pattern. In conclusion, sarcomas in general commonly express constitutive NOS, and only a few types of sarcomas can express iNOS, the isoenzymes capable of releasing large amounts of NO. More comprehensive studies should be performed to better understand the clinical importance of NOS expression and NO production in sarcomas. PMID- 17122637 TI - Alpha-methylacyl-CoA racemase (AMACR/P504S) can distinguish hepatocellular carcinoma and dysplastic hepatocytes from benign nondysplastic hepatocytes. AB - Immunohistochemical staining with alpha-methylacyl-CoA racemase AMACR (P504S) has been described in a number of normal tissues and was found to be useful for detecting malignancies including hepatocellular carcinoma (HCC). Our aim was to determine whether AMACR is differentially expressed in benign nondysplastic liver tissue, hepatocellular dysplasia, and HCC. The study material consisted of paraffin blocks containing primary HCC and surrounding liver tissue from 20 patients who underwent hepatectomy at the time of liver transplantation. Immunohistochemical stains were performed with anti-AMACR by standard methods. Staining features were characterized on the basis of the pattern and distribution of reactivity. A positive AMACR immunostain was defined as either finely stippled or coarsely granular in pattern, in a diffuse or parabasal cytoplasmic distribution. A negative AMACR immunostain was defined as absence of reactivity. Anti-AMACR immunostains were positive in malignant, dysplastic, and benign nondyplastic hepatocytes in all cases. The staining pattern was the same in malignant and dysplastic hepatocytes. It consisted of coarsely granular reactivity in a parabasal or diffuse cytoplasmic distribution. In contrast, benign nondysplastic hepatocytes were distinguished by weak, finely stippled diffuse cytoplasmic staining. Malignant and dysplastic hepatocytes showed an identical pattern of immunostaining for AMACR that was distinct from benign hepatocytes. Prospective studies are needed to determine whether staining for AMACR can distinguish HCC or dysplasia in cytologic and small histologic specimens. PMID- 17122638 TI - Immunohistochemical expression of Bcl-2, Bcl-x, and Bax in follicular carcinomas of the thyroid. AB - Bcl-2, bcl-x, and bax proteins are involved in the regulation of apoptosis. There is limited data on the expression of these proteins in follicular carcinomas (FCs) of the thyroid. A retrospective clinicopathologic review with bcl-2, bcl-x, and bax immunostaining of 34 FCs and 7 follicular adenomas with incomplete capsular penetration (FAICP) was performed. The study included 41 patients (25 females; age range 16 to 84 y, mean 50.9 y). All patients underwent surgical resection. Seven FC patients developed recurrent disease: 1 patient was alive (14.2 y) and 6 patients died with metastatic disease (mean survival 5.9 y). All remaining patients were disease-free (mean follow-up 7.9 y). Only one FAICP recurred (patient alive at 11 y). The remaining patients were disease-free (mean follow-up 6.9 y). Normal thyroid tissue stained positively for bcl-2 and bcl-x, and did not stain with bax. Only 15 tumors (12 FC and 3 FAICP) stained positively for bcl-2. None of the recurrent tumors demonstrated evidence of bcl-2 staining. The majority of tumors stained positively for bax (83%, 29 FC and 5 FAICP) and for bcl-x (93%, 32 FC and 6 FAICP); there was no correlation of staining with outcome. The majority of follicular neoplasms were positive for bax and negative for bcl-2 by immunohistochemistry. Aberrant expression of apoptosis-associated proteins may play a role in the pathogenesis of FC of the thyroid. All recurrent and fatal tumors were negative for bcl-2. These data suggest that the loss of bcl 2 expression may correlate with poorer prognosis. PMID- 17122639 TI - Immunohistochemical analysis of survivin expression in thyroid follicular adenoma and carcinoma. AB - Survivin is one of the 8 members of human inhibitor of apoptosis , which is differentially expressed in cancerous/transformed cells versus normal differentiated tissues. This retrospective study of thyroid histologic samples aimed to assess the clinical usefulness of survivin immunostaining for discrimination between follicular adenoma and carcinoma of thyroid. Immunohistochemical staining for survivin was performed on 41 lesions from patients who had undergone surgery for either follicular adenoma or carcinoma of thyroid. Survivin expression was significantly (P < 0.005) higher in the cases that received a diagnosis of carcinoma in comparison with follicular adenomas cases. Odds ratio of follicular carcinoma for survivin expression was 21.375 (95% CI: 3.283 to 139.177). Our results showed potential value of survivin in discrimination between follicular thyroid adenoma and follicular thyroid carcinoma. We conclude that survivin is a potential candidate for further investigation in the proper histologic diagnosis of thyroid cancers. PMID- 17122640 TI - Relationship between histologic features of primary breast carcinomas and axillary lymph node micrometastases: Detection and prognostic significance. AB - The incidence and prognostic significance of micrometastases (Mic-Met) in axillary lymph nodes (LNs) is still controversial. We compared Mic-Met detection of invasive mammary carcinomas (IMCs) in axillary LNs using second review of hematoxylin and eosin (H&E)-stained slides and immunohistochemistry (IHC) relating them with features of the primary tumor, and determining their influence on overall survival (OS) and disease-free survival (DFS). We studied 188 cases of IMCs with no axillary metastases in the initial reports. The original H&E slides of LN were re-viewed and new sections were submitted for IHC using pancytokeratin (AE1/AE3). All primary breast tumors were re-viewed and classified according to Page et al (1998) and College of American Pathologists criteria (2000). Tumors were graded using the Nottingham grading system. Kaplan-Meier curves were used to evaluate OS and DFS of 147 patients. Mic-Met detection was correlated to histologic features of primary tumor (size, type, grade, lymphatic/blood vessel invasion). Mic-Met were detected in 26/188 cases (by IHC: 23/188, 12.2%; by H&E: 12/188, 6.4%). The re-view of H&E slides showed good specificity (98.2%), but low sensitivity (39.1%), when compared with IHC. There was no relationship between features of primary tumor and Mic-Met detection, including patients with lobular carcinomas or IMCs with lobular features. There was no statistical difference in OS and DFS of patients with and without Mic-Met, but patients with Mic-Met presented lower survival curves. In conclusion, there was no relationship between histologic features of primary tumor and presence of Mic-Met, nor between Mic-Met detection and patients survival. PMID- 17122641 TI - Human Papillomavirus DNA and P16INK4A are not detected in renal tumors with immunohistochemistry and signal-amplified in situ hybridization in paraffin embedded tissue. AB - The association between human papillomavirus (HPV) infection and carcinogenesis has long been established in literature, with the strongest evidence for its role in cervical carcinoma. The role of HPV in urological tumors has been investigated and sporadic reports have linked HPV infection to bladder, prostate, renal, penile, and testicular cancer. Although less rigorously studied, there are a few conflicting results about the role of HPV in the development of malignant renal tumors. Moreover, no data are available for association of HPV DNA and expression of P16 in benign renal tumors. Formalin-fixed, paraffin-embedded tissues from 62 renal tumors (40 clear cell, 9 papillary, and 3 chromophobe renal cell carcinomas, 1 collecting duct carcinoma, 2 urothelial carcinoma of renal pelvis and 7 oncocytomas) were immunostained with low-risk and high-risk HPV DNA (6, 11, 16, 18, 31, 33, 42, 51, 52, 56, 58). Tissue microarray sections of 62 tumors were stained with P16 by immunohistochemistry. Signal amplified colorimetric in situ hybridization was performed on microarray sections using biotinylated probes for HPV subtypes 6, 11, 16, 18. A nuclear dot-like signal was considered positive for low-risk and high-risk HPV by immunohistochemistry and in situ hybridization and nuclear or cytoplasmic staining is considered positive for P16. No staining for HPV DNA and P16 was found in any type of renal tumors. Our results support that HPV does not seem to play a role in the development of benign and malignant renal tumors. PMID- 17122642 TI - Automation of manual components and image quantification of direct dual label fluorescence in situ hybridization (FISH) for HER2 gene amplification: A feasibility study. AB - Determination of HER2 status by fluorescence in situ hybridization (FISH) in breast carcinoma correlates well with response to targeted therapy and prognosis. However, manual time consuming methods and quantification aspects of the procedure may be challenging for some laboratories. We examined the feasibility of automating these components of the FISH assay using a tissue microarray (TMA 118 clinically annotated cases) and a series of 41 whole sections. An in situ hybridization automated staining workstation was used to automate a programmed overnight start, on line baking, deparaffinization, cell conditioning, protease digestion, and prehybridization buffer washing. Dual label probe/target codenaturation/hybridization and stringency washing were done off line. The HER2 and CEP17 spot counts were quantified, and the HER2/CEP17 ratio calculated, via an imaging workstation. Results were benchmarked against manual counts for whole sections, and bright field in situ hybridization [silver in situ hybridization (SISH)] for the TMA. Automated FISH results using whole sections correlated well with manual results: HER2/CEP17 ratio correlation coefficient r = 0.9154, r = 0.8380, P < 0.0001. Correlation between automated and manual TMA FISH results was also excellent, and disease-free survival was significantly shorter (P < 0.001) for the HER2 amplified cases. Automation of the laborious manual prehybridization and image quantification components of FISH using directly labeled probes is feasible. Operational gains and enhanced consistency are inherent in this automated approach to HER2 clinical FISH testing. PMID- 17122643 TI - Albumin in immunohistochemistry: Foe and friend. AB - Immunohistochemical procedures constitute a high methodological value in both pathologic diagnostics and research. Staining quality depends on a large variety of interference factors. Primarily, background staining reduces the quality of evaluation by reducing the chromatic discrimination. For the identification of important interference factors, various incubation steps and composition of solutions recommended in routine protocols were altered or omitted in our study. Surprisingly, the most important effect concerning background staining reduction could be significantly attributed to the omission of albumin which usually is recommended as a reducer of background stainings. However, in contrast to this negative effect, albumin could also increase specific staining intensity. These findings lead to the recommendation of a careful use of albumin in immunohistochemistry because of the dichotomous effects mentioned above. Furthermore, these results imply that in case of a good specific staining pattern, the use of albumin in immunohistochemical solutions merely exerts significant negative background staining effects. PMID- 17122644 TI - Solid-pseudopapillary neoplasm of the pancreas: Three cases with a literature review. AB - Solid-pseudopapillary neoplasm of the pancreas is a very rare tumor. It most commonly occurs in young women and has unique pathologic features. Previous immunohistochemical studies demonstrated that most solid-pseudopapillary neoplasms were immunoreactive with antibodies directed against vimentin and neuron-specific enolase. Recently, expression of CD10 and CD56 in this tumor has been reported. In this report, we expanded the demographic profile, highlighting 3 cases of solid-pseudopapillary neoplasm of the pancreas that presented in an elderly woman, a young man, and a young woman and further characterized them histologically and immunophenotypically. Grossly, all 3 tumors were well circumscribed and had a variable degree of cystic formation, necrosis, and hemorrhage. Microscopically, these tumors were characterized by a pseudopapillary pattern of epithelioid cells arranged around a delicate fibrovascular core with sheets of bland epithelioid cells filling cystic spaces. Hyaline globules, cholesterol granulomas, and foamy cells were all seen to be common findings. Although these 3 tumors were strongly immunoreactive for vimentin, alpha-1 antitrypsin, alpha-1-antichymotrypsin, neuron-specific enolase, CD10, CD56, and progesterone receptor, they demonstrated only variable "positivity" for epithelial membrane antigen and broad-spectrum cytokeratin, but were being consistently nonreactive for synaptophysin, insulin, glucogon, chromogranin A, and estrogen receptor. Interestingly, 2 of the 3 tumors were S-100 protein and melanin A reactive but were nonreactive for HMB45. PMID- 17122645 TI - Multiple immunofluorescence technology. PMID- 17122646 TI - The clinical significance of circulating tumor cells in the peripheral blood. AB - Tumors launch malignant cells into the circulation continuously. In early stages, the immune surveillance system eliminates these cells from the circulation, but at later times they may persist longer and be detected. The first recorded evidence of the presence of circulating tumor cells in the peripheral blood of cancer patients was documented in 1869. Now, modern molecular biologic and cell sorting techniques make their detection and characterization more practicable. This review will consider the methods currently available for their detection and characterization, and the clinical implications of their presence in various malignant conditions. PMID- 17122647 TI - Microarray analysis of MRI-defined tissue samples in glioblastoma reveals differences in regional expression of therapeutic targets. AB - Microarray technologies have come into prominence for the assessment of molecular diagnostic profiles in cancer tissue biopsies. To better understand the effect of sampling bias, we paired image-guided stereotactic biopsy and microarray technology to study regional intratumoral differences in tumor periphery and core regions of untreated glioblastoma. RNA was extracted from serial frozen sections using an integral histopathologic scoring approach. Gene expression analysis was performed using high-density oligonucleotide microarrays (22,283 probe sets). A consensus list of 643 genes (784 probe sets) with greater than 2-fold difference between intratumoral periphery and core samples was obtained using Microarray Suite 5.0, model-based expression indexes, and robust multiarray analysis algorithms. Results were validated using quantitative polymerase chain reaction and Western blotting analyses. Reproducible profiles emerged, in which multiple therapeutic targets significant to glioblastoma [matrix metalloproteinases, AKT1 (v-akt murine thymoma viral oncogene homolog 1), epidermal growth factor receptor, vascular endothelial growth factor] showed significant differences in regional expression that may affect treatment response. This study suggests important intratumoral regional differences in the molecular phenotype of glioblastoma. PMID- 17122648 TI - Pattern and distribution of immunoglobulin VH gene usage in a cohort of B-CLL patients from a Northeastern region of Italy. AB - We analyzed individual VH gene rearrangements in 55 consecutive B-chronic lymphocytic leukemia (B-CLL) patients collected from a northeastern region of Italy, stressing the possible differences related to geographic characteristics of the cohorts studied. Considering the percentage of somatic mutations present in the VH gene sequences and using the 98% cut-off value, 38 of the 55 B-CLL (69%) patients displayed somatic hypermutations and 17 (31%) had a germline configuration. Our results confirm and extend the observations of a bias in the use of certain VH, DH, and JH genes among B-CLL cells. The most frequently used VH genes were VH1-69 (12.7%) with VH3-23 (12.7%) and VH4-34 (10.9%). Collectively these genes accounted for 36.3% of the cases. In the mutated cases, the range of mutations varied from 2% to 15%, with a median of 6.5%. VH1-69 (7 cases, all unmutated) carried few mutations as opposed to VH3-23 (7 cases, 5 of which mutated), VH4-34 (6 cases, all mutated), and VH3-30 (5 cases, all mutated), which show a high load of mutations. D3 family genes were found frequently (38.1%) followed by D2 (27.2%) and D6 (18.1%). The individual D segment most frequently used was D3-3, which was present in 16.3% of cases. There was predominance of the JH4 gene (49%) followed by JH6 (40%). Analysis of the distribution of replacement and silent mutations in the mutated sequences using the method of Lossos showed in 39.4% of cases evidence of antigen selection in the framework region and/or complementary determining regions. In comparison with a recent study on B-CLL patients from the Mediterranean area, the VH4-34 gene was significantly overused in the mutated group at a percentage double that of the Italian cohort reported in this study (10.9% vs. 5%), but at a frequency similar to the entire Mediterranean region (10.7%). We also found an over-representation of VH1-69 usage in the germline group, at a frequency (12.7%) higher than previously described by the same authors (Italian 8%, Mediterranean 10.7%). On the contrary, VH3-07 and VH3-49 were not much used in our study (5.4% and 1.8%, respectively) compared with the Italian group (8% and 5.1%). In our study, VH3-23 gene segment was frequently expressed, at frequency as high as that of VH1-69, a finding in keeping with reported B-CLL Italian data, but higher than the entire series of the Mediterranean area (12.7% vs. 9.2%); VH3-21 gene, frequently expressed in northern European CLL but rarely in the Mediterranean area, was completely absent. This biased usage of VH family genes may reflect a geographic leukemic repertoire, perhaps owing to a peculiar genetic background, depending on variations in germline composition of the IgVH locus or to the effect of a potential environmental element less frequently encountered in different regions. PMID- 17122649 TI - Chromosomal and microsatellite instability of adenocarcinomas and dysplastic lesions (DALM) in ulcerative colitis. AB - Longstanding ulcerative colitis (UC) is associated with a high risk of developing UC-related colonic adenocarcinoma (UCC). These carcinomas originate from nonadenomatous dysplastic regions referred to as dysplasia associated lesion or mass (DALM). We evaluated chromosomal and microsatellite instability (MSI) in 21 DALM/UCCs. Chromosomal instability was determined by high-resolution array comparative genomic hybridization with a 3500-element BAC-PAC array. MSI was assessed with markers BAT25 and BAT26 and by immunohistochemical analysis of mismatch repair genes. Comparative genomic hybridization revealed frequent losses of array clones (>20% of tumors) at chromosome arms 4p, 5q, and 18q, frequent gains of array clones (>20% of tumors) were found at 1q, 5p, 6p, 7p, 7q, 8p, 8q, 11p, 11q, 12q, 14q, 17q, 19q, 20p, and 20q. The pattern of alterations is dominated by gains on 5p and 20q with loss of 4p, all of which were already present in a patient with carcinoma in situ. Immunohistochemical analysis of mismatch repair genes MLH1, PMS2, MSH2, and MSH6 showed negative immunostaining in 1 neoplasm (5%). MSI of BAT25 and BAT26 was seen in 3 tumors (14%) including the neoplasm with aberrant immunostaining. In conclusion, we constructed a genomic profile of DALM/UCC including several novel genetic alterations. Further, we found a low percentage of MSI. Thus, DALM/UCCs display profound chromosomal instability, but this is not associated with concurrent MSI. PMID- 17122650 TI - Histologic, viral, and molecular correlates of dengue fever infection of the liver using highly sensitive immunohistochemistry. AB - The mechanism by which the virus associated with dengue fever can cause a fatal hepatitis is not well understood. The purpose of this study was to examine 9 cases of fatal dengue hemorrhagic fever-associated hepatitis, and to correlate the histologic findings with viral detection and cytokine response. The histologic changes were nonspecific and included massive hepatic necrosis and a pauci-cellular acute hepatitis. Viral cDNA detection by reverse transcriptase in situ polymerase chain reaction demonstrated that the fatal hepatitis was due to infection on average of >90% of hepatocytes and many Kupffer cells. Similar results were obtained using immunohistochemistry for viral protein using an automated highly sensitive system. Immunohistochemical analysis for tumor necrosis factor alpha, and interleukin-2, showed rare positive Kupffer cells. In comparison, fatal cases of hepatitis C associated liver failure demonstrated far fewer infected hepatocytes and a concomitant strong up-regulation of many cytokines, notably tumor necrosis factor alpha and interleukin-2. It is concluded that fatal dengue hemorrhagic fever is associated with acute, severe liver damage due primarily to massive direct infection of hepatocytes and Kupffer cells with minimal cytokine response. The infection can be readily detected in a few hours using an automated system that has a sensitivity equivalent to reverse transcriptase in situ polymerase chain reaction. PMID- 17122651 TI - Optimization of recovery of RNA from formalin-fixed, paraffin-embedded tissue. AB - Formalin-fixed, paraffin-embedded (FFPE) tissue is the most common specimen available for application of diagnostic assays on tissue after microscopic examination. Not only is there a substantial archive of tissue available, but FFPE tissue remains the best method of preparation for microscopic examination in a routine clinical environment. Molecular assays, especially reverse transcription and polymerase chain reaction and expression array-based assays, offer significant potential as diagnostic, prognostic, and predictive tools, but require high quality RNA. Herein, we have optimized a reliable RNA extraction method for FFPE tissue. It is based on deparaffinization at high temperature coupled with a 3-day lysis at 65 degrees C. The average total RNA yield is 4.5 to 5.5 pg per 1 microm of archival FFPE tissue, and 260/280 ratios are between 1.80 and 1.95. The extracted RNA has a modal fragment length between 100 and 200 nt by the Bioanalyzer analysis. Although modal lengths of RNA fragments were shorter, reverse transcription and polymerase chain reaction was able to amplify amplicons in range of 300 bp. Pretreatment with RNA, later followed by formalin fixation, did not result in improving the RNA quality, but did improve RNA yield. Our method improves the utility of FFPE tissue for molecular profiling studies. PMID- 17122652 TI - Tissue pretreatment with formic acid might lower HercepTest scores in breast cancer. AB - Creutzfeldt-Jakob disease and other prion diseases are diseases with yet not well defined routes of transmission and infection. The safe processing of potentially contaminated tissue material remains a challenge for histologic laboratories. Formic acid pretreatment is considered to be effective in prion inactivation. We evaluated the c-erbB2 and the hormone receptor-status in potentially prion infectious breast cancer tissue after pretreatment with formic acid. Paired breast cancer tissue samples were immunostained with commercially available antibodies against c-erbB2, estrogen receptor, and progesterone receptor with 1 tissue sample of each pair being pretreated with 98% formic acid. Staining was evaluated either according to the HercepTest score or using an immunoreactive score. Additionally, fluorescence in situ hybridization (FISH) analyses were performed for 7 of these cases. Untreated tissues showed strong circumferential staining for c-erbB2 (HercepTest score 3+), whereas the membranous staining of the tissues pretreated with formic acid was significantly weaker. FISH analyses showed no differences in both groups. The hormone receptor expression was not significantly influenced and positivity was maintained in all cases. In breast cancer patients, the pretreatment of tissue with formic acid for prion decontamination in the case of suspected Creutzfeldt-Jakob disease or other prion diseases can lead to underestimation of the immunohistologically determined c erbB2 status. In these cases, a c-erbB2-FISH analysis should be performed. For the immunostaining of hormone receptors in breast cancer, formic acid pretreatment can be applied without negative effects on the sensitivity or specificity of the assay. PMID- 17122653 TI - Broken hearts: differentiating stress-induced cardiomyopathy from acute myocardial infarction in the patient presenting with acute coronary syndrome. AB - The link between an emotional or physical stresses and an acute cardiac event was first discovered in Japan. Stress-induced cardiomyopathy is defined as transient left ventricular dysfunction associated with apical and mid ventricular contractile abnormalities and sparing of basal segments that mimics acute myocardial infarction. This article presents an overview of this disorder. PMID- 17122655 TI - The challenges of managing and treating Guillain-Barre syndrome during the acute phase. AB - Guillain-Barre syndrome (GBS) is a randomly acquired inflammatory disease that affects approximately 2 persons in 100,000 annually. There have been no discriminating risk factors identified including age, sex, or race. The syndrome results in the demyelination of peripheral nerves, which leads to progressive motor weakness and paralysis. The critical care nurse should gain from this article an overview of Guillain-Barre syndrome during the acute phase. Included is the pathophysiology of the syndrome, clinical presentation, acute phase nursing assessment and management, and currently available treatment options. PMID- 17122656 TI - An overview of asthma. AB - Asthma is a disease characterized by reversible airflow obstruction and airway hyperresponsiveness. This disease accounts for thousands of hospitalizations each year. Critical care nurses play an important role in the management of acute asthma and in the prevention of acute exacerbations. This article presents an overview of asthma, its pathophysiology, signs and symptoms, and management. PMID- 17122657 TI - Should we allow children to visit ill parents in intensive care units? AB - Visitation policies in intensive care units are very strict. These serve to protect the critically ill patient. Should children be allowed to visit an ill parent or sibling? The developmental status of each child should be considered. At all times, children should be accompanied by another family member during their visit. Nurses are invited to send in their own comments and to provide their experiences regarding having children visit patients in intensive care units. PMID- 17122659 TI - Identification of special care needs: the comparison of the cardiothoracic intensive care unit patient and nurse. AB - The purposes of this study were to assess (a) the critically ill patient's and critical care nurse's identification and perception of special care needs and (b) the relationship between the patient's and nurse's perception of the identified special care needs. A qualitative approach, using linked data and open-ended surveys, was used to identify common themes and patterns in data. Data collection took place over a 2-month period at a metropolitan research hospital in the Southeast. Common themes and patterns of identified special care needs were recognized across data and with linked data between the patient and nurse. Findings indicate that both nurses and patients feel that the special care needs of the patient are being met. Common needs currently identified by the patient include miscommunication, anxiety, pain, and relaxation. Common needs currently identified by the nurses include pain management, nausea, and comfort. PMID- 17122668 TI - The evolving care of the elderly with heart failure: from the 'high-tech' to the 'high-touch' approach. PMID- 17122669 TI - Outcome of diabetic and non-diabetic patients undergoing successful coronary angioplasty with bare stent of chronic total occlusion. AB - BACKGROUND: The prognosis of patients with diabetes mellitus and chronic coronary total occlusion (CTO) treated with percutaneous coronary angioplasty (PTCA) is poorly investigated. METHODS: To compare the long-term outcome of patients with CTO, with and without diabetes mellitus and undergoing successful PTCA with bare stent implantation performed in a single centre, 170 consecutive patients (mean age 62 +/- 10 years) with CTO aged > 1 month were analysed. Death, myocardial infarction, repeat angioplasty and coronary artery by-pass were considered as hard events in 167 patients with available long-term follow-up (mean 25 +/- 15 months). RESULTS: Vessel mean luminal diameter after the procedure and stent length were 2.5 +/- 0.4 mm and 21.9 +/- 9.4 mm, respectively. No differences were found in baseline clinical, angiographic and procedural variables between the groups, categorized on the basis of presence or absence of diabetes. There were 13 (27%) and 25 (21%) events in diabetic and non-diabetic groups, respectively (P = not significant). Multivariate analysis identified final mean luminal diameter (odds ratio = 4.7192, P = 0.0013) and stent length (odds ratio = 1.0655, P = 0.0003) but not diabetes (P = 0.78) as predictors of events at long-term follow up. CONCLUSIONS: Patients with and without diabetes undergoing CTO re-opening with stent implantation do not differ at long-term follow-up in terms of death, myocardial infarction and target lesion revascularizations. Final mean luminal diameter and stent length are significant predictors of events during long-term follow-up. PMID- 17122670 TI - Effects of volume loading on strain rate and tissue Doppler velocity imaging in patients with idiopathic dilated cardiomyopathy. AB - BACKGROUND: Strain rate is a promising echocardiographic technique which adds further information to that obtained with two-dimensional echocardiography and tissue Doppler imaging (TDI). The present study aimed to evaluate the effects of acute isotonic volume expansion on left ventricular function in patients with idiopathic dilated cardiomyopathy (DCM) utilizing TDI and strain rate measurements. METHODS: Ten patients with DCM and a left ventricular ejection fraction (LVEF) or =4 visits). CONCLUSION: Rates of recognition of comorbid anxiety and depressive disorders are low in youth with asthma and few youth with asthma and comorbid anxiety and depression receive guideline-level mental health treatment. PMID- 17122710 TI - The effects of patient volume on the quality of diabetic care for Medicare beneficiaries. AB - BACKGROUND: The quality of care for Medicare beneficiaries with diabetes remains suboptimal. The contributing factors at the physician level are not well characterized, especially the relationship of patient volume and physician performance. OBJECTIVE: We sought to determine associations between the number of Medicare diabetic patients cared for by a primary care physician and the receipt of important diabetic processes of care. DESIGN: Physicians were grouped into quintiles based on the number of Medicare patients with diabetes. Hierarchical generalized linear models were used to examine associations between number of patients, frequency of visits, physician experience, patient factors and the receipt of diabetes processes of care using Part A and B Medicare claims data for 2001. PARTICIPANTS AND PATIENTS: All Connecticut primary care physicians who cared for Medicare fee-for-service beneficiaries with diabetes in 2001. MAIN OUTCOME MEASURES: The main outcome measures were associations of the receipt of diabetes process of care measures with the number of diabetic Medicare patients in a physician practice panel, adjusted for frequency of visits, patient comorbidity, age, ethnicity, and physician experience. RESULTS: Patients in the highest volume physician quintile were significantly more likely to have received hemoglobin A1c measurements, lipid profiles, and retinal eye examinations than patients in the lowest physician quintile in 2001, even after adjustment for multiple factors. For each step up in quintile volume group among primary care physicians, the increased odds of receiving a hemoglobin A1c measurement was 1.16 (95% confidence interval [CI] 1.10-1.23), 1.12 (95% CI 1.07-1.18) for a lipid profile, 1.06 (95% CI 1.02-1.09) for a retinal eye examination, and 1.48 (95% CI 1.22-1.81) for receiving all 3 measures. CONCLUSIONS: This study suggests that Medicare fee-for-service patients with diabetes cared for by physicians with greater numbers of diabetic Medicare patients in their practice are more likely to receive important diabetes processes of care. PMID- 17122711 TI - The effect of complementary and alternative medicine claims on risk adjustment. AB - OBJECTIVE: We sought to assess how the inclusion of claims from complementary and alternative medicine (CAM) providers affects measures of morbidity burden and expectations of health care resource use for insured patients. METHODS: Claims data from Washington State were used to create 2 versions of a case-mix index. One version included claims from all provider types; the second version omitted claims from CAM providers who are covered under commercial insurance. Expected resource use was also calculated. The distribution of expected and actual resource use was then compared for the 2 indices. RESULTS: Inclusion of claims from CAM providers shifted 19,650 (32%) CAM users into higher morbidity categories. When morbidity categories were defined using claims from all providers, CAM users in the highest morbidity category had average (+/-SD) annual expenditures of $6661 (+/-$13,863). This was less than those in the highest morbidity category when CAM provider claims were not included in the index ($8562 +/- $16,354), and was also lower than the highest morbidity patients who did not use any CAM services ($8419 +/- $18,885). CONCLUSIONS: Inclusion of services from CAM providers under third-party payment increases risk scores for their patients but expectations of costs for this group are lower than expected had costs been estimated based only on services from traditional providers. Risk adjustment indices may need recalibration when adding services from provider groups not included in the development of the index. PMID- 17122712 TI - Pediatrician coordination of care for children with mental illnesses. AB - BACKGROUND: Children with mental illnesses are seen across various service sectors, including pediatric primary care. As such, care coordination, which has been shown to improve outcomes, is especially important. Little is known about organizational and state level factors that might affect pediatricians' efforts to coordinate care for children with mental illnesses. OBJECTIVES: This study used data from a survey of primary care pediatricians to examine organizational and state level variables associated with increased care coordination for children with mental illnesses. METHODS: We undertook a cross-sectional study using data from a survey of pediatricians. A total of 1337 pediatricians in 6 states were surveyed about their care coordination and about organizational characteristics. State level data were gathered from various sources. A 5-item scale was developed for the dependent variable of past month coordination contacts (Cronbach's alpha = 0.82). RESULTS: No state level variables were associated with pediatrician coordination behaviors. Having a regular case conferencing mechanism, staff assigned to coordinate care, and a mental health specialist in the practice were associated with increased past month coordination contacts. Pediatricians in rural practices coordinated care more frequently than those in other locations. Pediatricians who screened more and those with more experience had higher rates of care coordination. CONCLUSIONS: Having a regular case conferencing mechanism, staff assigned to coordinate care, and a mental health specialist in the practice were associated with increased mental health care coordination. This information should be useful in planning ways to increase care coordination for children with mental illnesses seen in pediatric primary care. PMID- 17122713 TI - Connoisseurs of care? Unannounced standardized patients' ratings of physicians. AB - BACKGROUND: Patient satisfaction surveys can be informative, but bias and poor response rates may limit their utility as stable measures of physician performance. Using unannounced standardized patients (SPs) may overcome some of these limitations because their experience and training make them able judges of physician behavior. OBJECTIVES: We sought to understand the reliability of unannounced SPs in rating primary care physicians when covertly presenting as real patients. STUDY DESIGN: Data from 2 studies (Patient Centered Communication [PCC]; Social Influences in Practice [SIP]) were included. For the PCC study, 5 SPs made 192 visits to 96 physicians; for the SIP study, 18 SPs made 292 visits to 146 physicians. SPs visits to physicians were randomized, thus avoiding mutual selection bias. Each SP rated 16 to 38 physicians on interpersonal skills (autonomy support: PCC, SIP), technical skills (information gathering: SIP-only), and overall satisfaction (SIP-only). We evaluated SP evaluation consistency (physician vs. total variance rho), and SPs' overall satisfaction with specific dimensions of physician performance. RESULTS: Scale reliability varied from 0.71 to 0.92. Physician rhos (95% confidence intervals) for autonomy support were 0.40 (0.22-0.58; PCC) and 0.30 (0.14-0.45; SIP); information gathering rho was 0.46 (0.33-0.59; SIP). Overall SP satisfaction rho was 0.47 (0.34-0.60; SIP). SPs varied significantly in adjusted overall satisfaction levels, but not other dimensions. CONCLUSIONS: These analyses provide some evidence that medical connoisseurship can be learned. When adequately sampled by trained SPs, some physician skills can be reliably measured in community practice settings. PMID- 17122714 TI - Comparison of nurse, system and quality patient care outcomes in 8-hour and 12 hour shifts. AB - BACKGROUND: Many nurses desire 12-hour shifts. However, there are concerns about implementation. OBJECTIVE: We sought to compare the effects of 8- and 12-hour shifts on nurse, system, and quality patient care outcomes. METHODS: We used a cross-sectional design with data collected from multiple sources in 2003-2004, including a nurse survey and administrative and patient records. We studied hospital nurses and patients in general adult wards, with outcomes including burnout, job satisfaction, scheduling satisfaction, preferences, intention to stay, and employee safety. System outcomes included recruitment and turnover, staffing, absenteeism, and related costs. A variety of quality patient care outcomes were measured from the 3 different types of data. RESULTS: Thirteen New York City hospitals participated; 805 surveys were examined from 99 nursing units (response rate 42%). Compared with nurses working 8-hour shifts, those working 12 hour shifts were on average more satisfied with their jobs, experienced less emotional exhaustion, 10 times more likely to be satisfied with schedules, 2 times as likely to perceive 12-hour schedules as important, and 58% less likely to report missing shifts; units with 12-hour shifts had lower vacancy rates and weeks to fill the position (all P values < or =0.05). There were no differences in patient outcomes. CONCLUSIONS: Nurses working 12-hour shifts were more satisfied. There were no differences in quality outcomes. Flexibility and choice in shift length are important elements in a positive nurse work environment. This study represents an innovative attempt by a labor-management bargaining group to make an evidence-based decision. We encourage others to conduct similar studies. PMID- 17122715 TI - Do patient requests for antidepressants enhance or hinder physicians' evaluation of depression? A randomized controlled trial. AB - OBJECTIVE: We sought to ascertain whether patients' requests for antidepressants affect visit duration or history taking by primary care physicians (PCPs) for patients with depressive symptoms and a coexisting musculoskeletal disorder and to determine whether more thorough history taking is associated with diagnostic accuracy or with provision of minimally acceptable initial care for major depression. DESIGN: This was a randomized trial using standardized patients (SPs). Six roles involved 2 conditions (major depression and adjustment disorder, both with coexisting musculoskeletal conditions) and 3 patient request types (brand-specific, general, or none). We conducted the study in 152 PCP offices in Northern California and Rochester, New York. Physicians were assigned randomly to see 2 SPs with depression/wrist pain or adjustment disorder/back pain. MAIN OUTCOME MEASURES: Physician history-taking for depression and the musculoskeletal condition; depression diagnosis in the medical record; antidepressant prescriptions/samples; referral/follow-up recommendations; visit duration; and provision of minimally acceptable initial depression care. RESULTS: General antidepressant requests were associated with more depression history-taking (Adjusted Parameter Estimate = 0.80 more questions of 10 (95% confidence interval = 0.31-1.29, P < 0.001); brand-specific requests were marginally associated with more depression history-taking (Adjusted Parameter Estimate = 0.45, 95% confidence interval = -0.04-0.93, P = 0.07). Antidepressant medication requests were not related to musculo-skeletal question asking (P > 0.3) or visit length (P > 0.8). Depression history taking was directly associated with the likelihood of a chart diagnosis of depression and the provision of minimally acceptable initial depression care. CONCLUSION: General antidepressant requests increase depression history taking, including screening for suicide. Patients' requests for medication do not appear to short-circuit history taking for depression or distract the physician's attention from coexisting musculoskeletal conditions. PMID- 17122716 TI - Nursing working conditions in relation to restraint practices in long-term care units. AB - OBJECTIVES: This study examined the effects of nursing working conditions on the use of physical restraints and antipsychotics as restraints in long-term care units for elderly residents. DESIGN: Cross-sectional data were obtained in Finland in 2002 from long-term care units that used the Resident Assessment Instrument (RAI) system and participated in a survey on working conditions. SETTING: A sample of 91 inpatient units in 31 facilities (23 residential homes and 8 health centers). PARTICIPANTS: Data included 2430 resident assessments and 977 nursing staff survey responses. MEASUREMENTS: We measured unit-level mean scores of physical restraint and antipsychotics use as restraints and resident characteristics (activities of daily living, cognitive impairment, and daily behavioral problems) based on the RAI system as measured by the Minimum Data Set 2.0. Head nurses reported the structural factors (nurse staffing levels and unit size). Nursing working conditions were measured by the Job-Demands and Job Control Scales in the staff survey questionnaire. RESULTS: Controlling for resident characteristics, nurses' job demands and control had a combined effect on restraint practices. Job demands strongly increased the risk of physical restraint use in units where nurses reported low job control (odds ratio [OR] = 13.31, 95% confidence interval [CI] 1.55-114.30, P = 0.019), but not among high control units (OR = 0.23, 95% CI 0.04-1.29, P = 0.090). Although the use of antipsychotics was not related to job demands in units with low control (OR = 1.11, 95% CI 0.26-4.99, P = 0.891), the antipsychotics use in particular decreased when high job demands were coupled with high job control (OR = 0.17, 95% CI 0.03-0.91, P = 0.038). CONCLUSION: The results suggest that restraint use can be reduced by enhancing working conditions so that the nursing staff has possibilities for skill usage and decision-making. PMID- 17122717 TI - The association between clinical care strategies and the attenuation of racial/ethnic disparities in diabetes care: the Translating Research Into Action for Diabetes (TRIAD) Study. AB - OBJECTIVE: We sought to determine whether greater implementation of clinical care strategies in managed care is associated with attenuation of known racial/ethnic disparities in diabetes care. RESEARCH DESIGN AND METHODS: Using cross-sectional data, we examined the quality of diabetes care as measured by frequencies of process delivery as well as medication management of intermediate outcomes, for 7426 black, Latinos, Asian/Pacific Islanders, and white participants enrolled in 10 managed care plans within 63 provider groups. We stratified models by intensity of 3 clinical care strategies at the provider group level: physician reminders, physician feedback, or use of a diabetes registry. RESULTS: Exposure to clinical care strategy implementation at the provider group level varied by race and ethnicity, with <10% of black participants enrolled in provider groups in the highest-intensity quintile for physician feedback and <10% of both black and Asian/Pacific Islander participants enrolled in groups in the highest intensity quintile for diabetes registry use. Although disparities in care were confirmed, particularly for black relative to white subjects, we did not find a consistent pattern of disparity attenuation with increasing implementation intensity for either processes of care or medication management of intermediate outcomes. CONCLUSIONS: For the most part, high-intensity implementation of a diabetes registry, physician feedback, or physician reminders, 3 clinical care strategies similar to those used in many health care settings, are not associated with attenuation of known disparities of diabetes care in managed care. PMID- 17122718 TI - Redirecting patients to improve stroke outcomes: implications of a volume-based approach in one urban market. AB - BACKGROUND: The designation of primary stroke centers may result in patients being redirected from their usual source of care, although there is little evidence that these centers would result in better outcomes or lower costs. An alternative approach could direct patients to hospitals treating greater volumes of stroke patients. OBJECTIVES: We sought to estimate the effect of hospital stroke volume on patient mortality and costs in a regional hospital market and to analyze the implications of hypothetical volume-based referral policies in that market, including the effects of patient-hospital distance. METHODS: Using a retrospective cohort, we studied 12,150 Medicare patients admitted for acute stroke to 1 of 29 hospitals in Greater Cleveland during a 7-year period. The primary outcome was risk-adjusted 30-day mortality. Secondary outcomes included log hospital costs and discharge destination. The primary measure of volume was average annual number of stroke patients; patient distance to the nearest hospital was approximated using patient zip code and hospital address data. RESULTS: Overall 30-day mortality was 14.9%. For each 100-patient increase in hospitals' annual stroke volume, risk-adjusted mortality declined 0.9 percentage points (odds ratio = 0.90; 95% confidence interval = 0.82-0.98; P < 0.02) with no significant difference in hospital costs. For each 1-mile increase in patient distance to nearest hospital, mortality increased 0.6 percentage points (odds ratio = 1.07; 95% confidence interval = 1.03-1.11; P < 0.01). Only 3 of 29 hospitals (10.3%) exceeded the highest plausible threshold (250 strokes/year), redirecting 81.4% of patients for a net reduction in mortality of 0.4%; lower thresholds would redirect fewer patients but have negligible effects on mortality. CONCLUSIONS: Our findings fail to support redirecting acute stroke patients based on hospital stroke volume. PMID- 17122719 TI - The influence of patient race and social vulnerability on urologist treatment recommendations in localized prostate carcinoma. AB - BACKGROUND: In localized prostate carcinoma (PCa), many studies have found that black subjects receive radical prostatectomy (RP) less often than white subjects. Such disparities involve barriers to health care, comorbid illnesses, tumor characteristics, and patient preferences. It is unclear whether differences in urologist treatment recommendations also might play a role. METHODS: Using a randomized, 2 x 2 factorial design, we presented 2000 urologists with a clinical vignette and asked them to recommend treatment of a healthy 70-year-old patient with low-risk, clinically localized PCa. Options included either RP, external beam radiotherapy, brachytherapy, cryotherapy, observation, or hormonal therapy. There were 2 variables within 4 otherwise-identical versions of the vignette: 1) patient race (black vs. white) and 2) social vulnerability (middle-income and married vs. low-income and widowed). We used multivariable logistic regression to model the effects of patient race, social vulnerability, and their interaction on recommendations for RP versus radiotherapy. RESULTS: The response rate was 66.1% (n = 1313). Race and social vulnerability interacted (P = 0.05) such that the highly vulnerable black patient received an RP recommendation 14.4% less often than his less vulnerable counterpart; the difference between the 2 white patients was 4.2%. DISCUSSION: Race interacts with social vulnerability to influence urologist recommendations for RP. Because PCa tends to be more lethal in blacks, urologists may view such patients as good candidates for RP. However, black race may amplify perceptions of social vulnerability, heightening urologists' concerns about poor surgical outcomes and follow-up. These findings affirm the importance of modeling interactions between race/ethnicity and other social variables in health disparities research. PMID- 17122720 TI - Effect of quality improvement organization activities on outpatient diabetes care in eastern New York State. AB - BACKGROUND: Quality improvement organizations (QIOs) are contracted to improve the quality of care delivered to Medicare beneficiaries. The purpose of this study was to determine whether provider participation in New York State QIO activities resulted in significant improvements in the quality of diabetes care during the recent contract cycle with the Centers for Medicare & Medicaid Services. RESEARCH DESIGN: A retrospective analysis between participating and nonparticipating providers on their performance in 3 quality measures (biennial ophthalmology examination, biennial lipid profile monitoring, annual hemoglobin A1c monitoring) was used. Data of New York State Medicare beneficiaries before and after QIO intervention activities were examined to determine change in performance. General linear models were created to examine the effect QIO participation had on the change in performance for each measure. RESULTS: Providers who participated in QIO activities had significant absolute improvements in lipid monitoring compared with nonparticipating providers at high baseline performance for low (3.10%, P < 0.001), medium (2.57%, P < 0.001), and high (1.51%, P = 0.002) baseline patient volume, and medium baseline performance for low (2.38%, P < 0.001), and medium (1.85%, P < 0.001) baseline patient volume. The same trend was seen for hemoglobin A1c monitoring (4.28%, P < 0.001; 3.57%, P < 0.001; 2.15%, P < 0.001; 2.63%, P = 0.001; 1.92%, P = 0.006). For ophthalmology examination, participation resulted in significant changes at low (2.28%, P = 0.003) and medium (1.73%, P = 0.009) baseline patient volume. CONCLUSION: The study results suggest QIO activities can improve outpatient diabetes care; however, limitations in the study design preclude any definitive remarks. PMID- 17122721 TI - Response to an article in the April 2006 issue of Medical Care. PMID- 17122722 TI - Response to an Article in the June 2006 issue of Medical Care. PMID- 17122724 TI - Glioma therapy in adults. AB - BACKGROUND: Gliomas are the most common type of primary brain tumor. Nearly two thirds of gliomas are highly malignant lesions that account for a disproportionate share of brain tumor-related morbidity and mortality. Despite recent advances, two-year survival for glioblastoma with optimal therapy is less than 30%. Even among patients with low-grade gliomas that confer a relatively good prognosis, treatment is almost never curative. REVIEW SUMMARY: Surgery and radiation have been the mainstays of therapy for most glioma patients, but temozolomide chemotherapy has recently been proven to prolong overall survival in patients with glioblastoma. Intriguing data suggests that activity of O6 methylguanine-DNA methyltransferase (MGMT), in tumor cells may predict responsiveness to temozolomide and other alkylating agents. Novel treatment approaches, especially targeted molecular therapies against critical components of glioma signaling pathways, appear promising in preliminary studies. Optimal treatment for patients with low-grade gliomas has yet to be determined. Advances in oligodendroglioma biology have identified loss of chromosomes 1p and 19q as powerful indicators of a favorable prognosis. These same changes may predict response to chemotherapy. CONCLUSIONS: Though the prognosis for many patients with gliomas is poor, the last decade produced a number of important advances, some of which have translated directly into survival benefits. Rapid progress in the field of glioma molecular biology continues to identify therapeutic targets and provide hope for the future of this challenging disease. PMID- 17122725 TI - Review of progressive multifocal leukoencephalopathy and natalizumab. AB - BACKGROUND: Progressive multifocal leukoencephalopathy (PML), a destructive demyelinating infection which lytically infects oligodendrocytes, has occurred in patients treated with natalizumab. Magnetic resonance imaging (MRI) scan imaging of the brain gives clues to diagnosis but is nonspecific in distinguishing multiple sclerosis from PML. Spinal fluid detection of JC virus is specific but incompletely sensitive. Associated immunosuppression is typically of the cell mediated type but can be poorly defined on clinical grounds. REVIEW SUMMARY: It is apparent that natalizumab is a predisposing factor for developing PML from the 3 cases of natalizumab-treated patients. There is no reliable presymptomatic way to detect PML or JC virus infection of the brain by virologic or imaging surveillance techniques. One patient with multiple sclerosis and natalizumab treatment has survived, indicating that withdrawal of antibody, possibly in combination with antiviral therapy, may permit survival. However, immune reconstitution disease is a risk after immune restoration and withdrawal of natalizumab. PML deficits would be expected to be permanent. The estimate of incidence of PML in natalizumab-treated patients is 1 per 1000. The duration of natalizumab treatment may be an independent risk factor for development of PML. CONCLUSION: PML, a usually fatal neurologic infection, should be considered as a risk factor when using natalizumab. The treatment of multiple sclerosis patients with natalizumab is a matter of informed risk, individualized for each multiple sclerosis patient. PMID- 17122726 TI - Pediatric multiple sclerosis. AB - BACKGROUND: It is becoming ever more recognized that multiple sclerosis (MS) occurs in children and adolescents. Although early-onset MS is a variant in the spectrum of MS phenotypes, the diagnosis and management of MS in children poses a unique set of challenges for the clinician. Moreover, it is increasingly clear that these patients require specialized care, which includes a closely monitored medication program, physical therapy, cognitive and educational evaluation and intervention, and psychosocial support both for the patient and family. REVIEW SUMMARY: The goal of this article is to familiarize the clinician with the clinical and MRI features of pediatric multiple sclerosis and to summarize what is currently known about the clinical disease course. This article highlights important entities in the differential diagnosis of pediatric MS and discusses management strategies based on current literature. CONCLUSION: Children and adolescents with MS comprise a small but important subset of MS patients who require a multidisciplinary care approach. Additional multicenter studies are required to explore the effects of disease and treatments on physical, psychosocial, educational, and developmental parameters. PMID- 17122727 TI - Anaplastic astrocytoma presenting as reversible posterior leukoencephalopathy syndrome. AB - We report a 60-year-old man with grade III astrocytoma, who presented with status epilepticus. The initial MRI did not demonstrate typical findings of an astrocytoma but rather showed reversible posterior leukoencephalopathy syndrome (RPLS). N-Isopropil-p-[I] iodoamphetamine single photon emission computed tomography (SPECT) demonstrated hyperperfusion in this area. A brain tumor should be considered and the patient carefully followed by MRI, even if the MRI white matter lesion pattern suggests RPLS. This is especially relevant in the presence of atypical findings for RPLS on SPECT. PMID- 17122728 TI - Neurosyphilis presenting with status epilepticus. AB - BACKGROUND: With the introduction of antibiotics, neurosyphilis is now considered uncommon, and accurate diagnosis sometimes is problematic as the symptoms and signs are not pathognomonic. REVIEW SUMMARY: We present a 51-year-old man who had a 4-month history of general malaise and subtle cognitive impairment, combined with a normal brain image study, who ultimately developed an episode of status epilepticus as the main manifestation of unrecognized neurosyphilis. CONCLUSION: Because a clinical diagnosis of neurosyphilis is rarely encountered today in the developed world, a high index of suspicion and clinician awareness is very important in early diagnosis. PMID- 17122729 TI - Epilepsy and celiac disease: favorable outcome with a gluten-free diet in a patient refractory to antiepileptic drugs. AB - BACKGROUND: There is a well-documented relationship between epilepsy and celiac disease, including a syndrome characterized by epilepsy, occipital calcifications, and celiac disease. REVIEW SUMMARY: We report the case of a 23 year-old woman with an 11-year history of refractory epileptic seizures and newly diagnosed biopsy-proven celiac disease with increased antiendomysium immunoglobulin A antibodies. The patient showed a dramatic improvement after starting a gluten-free diet. CONCLUSION: This case emphasizes the need to include celiac disease in the differential diagnosis when investigating the etiology of epilepsy in refractory patients. PMID- 17122730 TI - A diagnostic and management dilemma: combined paraneoplastic myasthenia gravis and Lambert-Eaton myasthenic syndrome presenting as acute respiratory failure. AB - BACKGROUND: Neuromuscular junction disorders are usually categorized as either presynaptic or postsynaptic. The most frequently encountered disorder of the postsynaptic neuromuscular junction is acquired myasthenia gravis. Lambert-Eaton myasthenic syndrome is a well-known prototype of the presynaptic autoimmune disorders of neuromuscular transmission. These major disorders of neuromuscular transmission are relatively common and distinctly recognized, but co-occurrence of these disorders (overlap myasthenic syndrome) is rare and has so far attracted little attention. REVIEW SUMMARY: This report describes a patient with acquired myasthenia gravis and immunologic coexistence of Lambert-Eaton myasthenic syndrome (overlap myasthenic syndrome) in association with abdominal/uterine leiomyosarcoma. The patient presented with acute respiratory failure, making identification and management of her illness challenging. A general overview of the complexities associated with overlap between myasthenia gravis and Lambert Eaton myasthenic syndrome is provided and this patient's complicated clinical course and response to therapy are discussed. CONCLUSION: To our knowledge, this is the first report of overlap myasthenic syndrome in conjunction with abdominal leiomyosarcoma. The immunologic coexistence of acquired myasthenia gravis and Lambert-Eaton myasthenic syndrome in a patient with a malignant smooth-muscle tumor is intriguing and suggests that a common paraneoplastic process targeting 2 different onconeural antigens was the underlying pathogenic mechanism in this patient. PMID- 17122731 TI - Chorea-acanthocytosis: a mimicker of Huntington disease case report and review of the literature. AB - BACKGROUND: Neuroacanthocytosis consists of a group of rare heterogeneous neurodegenerative disorders associated with acanthocytosis. Chorea acanthocytosis, a variety of neuroacanthocytosis, is an autosomal recessive condition with clinical and radiologic features similar to Huntington disease. Although difficult, distinguishing between these entities is crucial as the implications for genetic counseling are significant. REVIEW SUMMARY: We report the case of a 33-year-old female who presented to our institution with a 3-year history of chorea. The patient's prominent orofacial symptoms and the presence of acanthocytes on peripheral blood smear led to the correct diagnosis of chorea acanthocytosis. CONCLUSIONS: The significant similarities between chorea acanthocytosis and Huntington disease at the clinical and radiologic levels can lead to an initial misdiagnosis. Clinical clues suggestive of chorea acanthocytosis include prominent orofacial dyskinesias, often causing dysarthria and dysphagia. Acanthocytosis, when present on peripheral blood smear, can confirm the diagnosis. PMID- 17122732 TI - Essential tremor. PMID- 17122736 TI - NPs a ray of hope for the mentally ill. PMID- 17122737 TI - Acute care billing: shared visits. PMID- 17122738 TI - Getting paid what you're owed. PMID- 17122739 TI - Heart failure epidemic boiling to the surface. PMID- 17122741 TI - Asthma pathogenesis and management. PMID- 17122743 TI - Exposing the dangers of anhydrous ammonia. PMID- 17122745 TI - Guide to care for patients. Asthma inhalers. PMID- 17122744 TI - Prescribing SSRIs for adolescents with depression. PMID- 17122746 TI - Nursing responsibilities in a disaster. PMID- 17122747 TI - The 3rd National Family History Initiative: Thanksgiving 2006. PMID- 17122751 TI - Metabolic syndrome and renal failure: similarities and differences. AB - The metabolic syndrome (MS) and chronic kidney disease (CKD) share many similar risk factors for cardiovascular disease. Both are associated with increased triglyceride (TG) levels, both associated with increased small dense low density lipoprotein (LDL), both with decreased high density lipoprotein (HDL) levels. In both cases HDL particle size is reduced. The TG content of HDL and very low density lipoprotein (VLDL) and remnants are increased, resulting in a dyslipidemia. Both are associated with increased inflammation, a hypercoagulable state and insulin resistance. Establishing whether these similarities are the result of identical biological processes or instead represent similar end results of different processes is further confounded by the associated both of adiposity and of MS with the incidence and progression of renal failure. Differences are present however. In MS hepatic VLDL synthesis is increased driven by increased flux of free fatty acids from muscle, adipose tissue and gut to the liver. VLDL is catabolized to LDL and the transfer of TG to HDL by cholesterol ester transfer protein destabilizes HDL leading to its rapid clearance. In CKD HDL fails to mature due to reduced activity of lecithin cholesterol transfer protein. In MS inflammation primarily arises from increased visceral adipose tissue, while inflammation is largely unrelated to body composition in CKD. Increased production of TG rich lipoproteins predominates in MS, while decreased disposal of TG rich proteins predominates as the cause of increased TG levels in CKD. Whether treatment of elements of MS, with the exception of hypertension, will avoid the onset and progression of renal failure is unknown. PMID- 17122752 TI - Liver cancer: a disease of stem cells? AB - Studies of aggregation chimaeras and X-linked polymorphisms strongly suggest that most liver tumours are derived from single cells (monoclonal): this review looks at the various cells in the liver that could be the founder cells for the two major primary tumours, namely hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC). In continually renewing tissues such as the gut mucosa and epidermis, where a steady flux of cells occurs from the stem cell zone to the terminally differentiated cells that are imminently to be lost, it is widely accepted that cancer is a disease of stem cells, since these are the only cells that persist in the tissue for a sufficient length of time to acquire the requisite number of genetic changes for neoplastic development. In the liver the existence of bipotential hepatic progenitor cells (HPCs), along with hepatocytes endowed with longevity and long-term repopulating potential suggests there may be more than one type of carcinogen target cell. Cell proliferation at the time of carcinogen exposure is essential for fixation' of any genotoxic injury into a heritable form, thus any proliferative cell in the liver can be susceptible to neoplastic transformation. Hepatocytes are implicated in many instances of HCC (e.g. in mouse models of HCC, oncogenic transgenes are driven by albumin promoters): chronic inflammatory injury to the biliary epithelium suggests cholangiocytes can give rise to CC, while HPC/oval cell activation accompanies many instances of liver damage irrespective of aetiology, making such cells likely carcinogen targets. Of course, we must qualify this assertion by stating that many carcinogens are both cytotoxic and cytostatic, and that HPC proliferation may be merely a bystander effect of this toxicity. PMID- 17122753 TI - Stem cells and Helicobacter-induced gastric cancer: where do we stand at the end of 2006. AB - The process of carcinogenesis, which may well extend over decades, provides an excellent opportunity for early detection and intervention to prevent development of gastric cancer. Evidence supporting a causal association between such tumour and Helicobacter infection has been demonstrated by epidemiological data, ecologic studies and, in experimental animal models. Lately, an increasing amount of evidences point out to bone marrow stem cells (BMSC) as target of neoplastic transformation. The term BMSC includes a heterogenous group of cells that both in vivo and in vitro studies, have shown to have plasticity, with their ability to acquire features of mesoderm, ectoderm and endoderm. In the gastric setting, a recent experiment in C57BL/6 mice has permitted to show that BMSC are the cell of origin of Helicobacter-induced gastric cancer. Based on these results, a model for epithelial cancer by which chronic inflammation leads to tissue injury and with time, to tissue stem cell failure, has been proposed. This phenomenon would induce the recruitment and engraftment of BMSC into the tissue stem cell niche. During differentiation, BMSC fail to regulate growth programme appropriately and progress through stages of metaplasia and dysplasia. Due to several reasons, in humans, data are conflicting. Further studies may shed light on the molecular bases of gastric lesions, leading to the development of preventive strategies. PMID- 17122754 TI - Geometry and bone density. AB - Bone development is a key process in the growing child. It is, therefore, of paramount importance to survey this process, which is characterized by increasing length and size of the bone together with its progressive mineralization. The bone status can be evaluated by different techniques, each of them having its pros and cons. Furthermore, it should be underlined that the results of bone assessment depend not only from the employed technique but also from the auxological characteristics of the subjects. It is, therefore, the aim of this review to examine the characteristics of the various methods of bone evaluation, such as dual energy X-ray absorptiometry (DEXA), peripheral quantitative computed tomography (pQCT), ultrasound and metacarpal index and to explain how changes in bone structure and geometry may influence the results. PMID- 17122755 TI - Transcranial Doppler for monitoring the cerebral blood flow dynamics: normal ranges in the Italian female population. AB - AIM: This study was aimed at recording, by means of a transcranial Doppler (TCD) device, the values of blood flow velocities (BFV) in the middle cerebral artery (MCA) in physiological conditions, in a population of healthy women in various ages of their life, in order to establish normative data in an Italian female population. METHODS: Our sample consisted of 100 healthy women (mean age 38 +/ 15.14 years, range 12-78) that underwent an investigation of the intracranial circulation by means of TCD. Patients were subdivided into 4 age groups: less than 20 years; 20-34 years; 35-50 years; more than 50 years. RESULTS: No statistically significant differences were present between the mean BFV in the left and right MCAs of the subjects considered as a unique population (left MCA 68.06+/-9.22 cm/s; right MCA 66.71+/-8.79 cm/s). The BFVs tended to significantly decrease with the increasing of age. In fact, they were so distributed: left MCA 82.55+/-6.85 cm/s and right MCA 80.27+/-4.13 cm/s in the younger group; left MCA 72.15+/-6.37 cm/s and right MCA 70.68+/-6.79 cm/s in women aged 20-34; left MCA 63.85+/-7.08 cm/s and right MCA 63.06+/-7.29 cm/s in women aged 35-50; left MCA 60.67+/-6.85 cm/s and right MCA 59.10+/-5.36 cm/s in the last group. CONCLUSIONS: The present study defined a normal age-related range of variations in MCA BFVs, useful for future comparisons in studies involving pathological female subjects. PMID- 17122756 TI - Identification of a novel mutation in the spastin gene (SPG4) in an Italian family with hereditary spastic paresis. AB - Hereditary spastic paraparesis (HSP) includes a heterogeneous group of neurodegenerative diseases characterised by progressive spasticity and hyper reflexia of the lower limbs. Autosomal dominant HSP type 4 is the most common clinical form, accounting for about 40-50% of autosomal dominant HSP families. This form is due to mutation of the gene encoding spastin (SPG4), an ATP-ase associated with a variety of cellular function (AAA). Here we describe a novel missense mutation (1297T>C; 391L>P) in exon 8 of SPG4 gene, identified in 2 members (mother and son) of an Italian family with autosomal dominant HSP, clinically pure in the mother and complicated in the son. The mutation lies in a highly conserved AAA box domain between amino acids 342 and 599 in spastin sequence. In both patients, this novel mutation was associated with the absence of relatively common clinical characteristics, such as vibratory sensory deficit and loss of sphincter control, and partial temporal epilepsy, particularly in the son, with infantile onset, secondarily generalised and moderately severe neuropsychiatric symptoms. PMID- 17122757 TI - Phlegmonous abscess associated with etanercept therapy. PMID- 17122758 TI - Which one is responsible for pain during transrectal guided prostate biopsy: prostate or anal canal. PMID- 17122759 TI - The concept of normality through history: a didactic review of features related to philosophy, statistics and medicine. AB - Normality characterises in medicine any possible qualitative or quantitative situation whose absence implies an illness or a state of abnormality. The illness concept was first a philosophical one. But the use of mathematics in the study of biological events, which began with Galton (1822-1911) and with Pearson (1857 1936), changed the frame of reference. In the second part of the 19th century mathematics was used to study the distribution of some biological characteristics in the evolution of the species. Around 1900, statistics became the basis for the study of the diffusion of the illnesses. Half a century later statistics made possible the transition from the description of single cases to groups of cases. Even more important is the concept of "normality" in laboratory medicine. In this field the search for the "perfect norm" was, and possibly still is, under way. The widespread use of statistics in the laboratory has allowed the definition, in a certain sense, of a new normality. This is the reason why the term "reference value" has been introduced. However, even the introduction of this new term has merely shifted the problem, and not resolved it. PMID- 17122760 TI - [Laparoscopic-assisted versus open surgery for colorectal cancer: postoperative morbidity in a single center randomized trial]. AB - AIM: The primary objective of the study was to compare the effect of laparoscopic assisted (LPS) versus open surgery (LPT) for colorectal cancer on postoperative morbidity. The secondary objectives were to evaluate immune response and intestinal wall oxygen perfusion. METHODS: A total of 610 patients with colorectal cancer were randomly assigned to receive colon resection by either LPS (n=306) or LPT (n=304). Four surgical staff members not involved in the study recorded postoperative complications up to 30 days after the operation. Immune response was evaluated by measuring lymphocytic proliferation after challenge with Candida albicans and phytohemoagglutinin before, at 3 and 15 days after the operation. Intestinal wall oxygen perfusion was continuously monitored using a probe. RESULTS: The conversion rate was 4.6% in the LPS group. Morbidity was 18.6% in the LPS group and 34.5% in the LPT group (P<0.0005). Infections developed in 9.1% of LPS-treated patients and in 20.2% of LPT-treated patients (P<0.0005). The mean length of stay was 9.7+/-2.6 days in the LPS group and 12.2+/-4.2 days in the LPT group (P<0.0001). In both groups lymphocytic proliferation was low at 3 days postoperative but returned to normal range at 15 days only in the LPS group. Interoperative intestinal oxygen perfusion values were higher in the LPS group. CONCLUSIONS: Laparoscopic colorectal surgery reduced both postoperative morbidity and length of stay. Lymphocytic proliferation and intestinal wall oxygen perfusion were higher in patients who underwent laparoscopic-assisted surgery. PMID- 17122761 TI - Standardized laparoscopic right hemicolectomy technique for colon cancer. AB - AIM: We analysed our experience of laparoscopic surgical treatment vs traditional surgery of right colon cancer. METHODS: A series of 27 patients was treated from January 2001 to December 2005 out of a total of 927 surgical colorectal operations in the same period (147 with the laparoscopic approach). Inclusion and exclusion criteria are reported. We compared this group with 25 patients treated by the same surgical group with open surgery. The mean operative time, the distance from the distal margin of resection, the number of lymphnodes, the mean period of canalization and the mean hospital stay are reported. RESULTS: In the laparoscopic group, the mean operative time was 124.8+/-36.3 min vs open surgery group of 94+/-23.6 min; the distance from the distal margin was 6.7+/-3.1 cm vs 6.4+/-2.1 cm; number of lymphnodes was 15.2+/-4.3 vs 18.7+/-2.9 nodes; and canalization 1.7+/-0.9 vs 2.7+/-0.7 days. The hospital stay was 6.8+/-1.7 vs 7.2+/-0.8 days. CONCLUSIONS: We consider laparoscopic right colon resection a safe procedure but it needs good laparoscopic practice and the observance of inclusion criteria. PMID- 17122762 TI - [Mid and long term results in the laparoscopic surgical treatment of gastroesophageal reflux]. AB - AIM: The advent of laparoscopic surgery produced a new interest in the surgical treatment of many benign and functional gastrointestinal diseases; one of the most commonly treated is the gastroesophageal reflux disease (GERD). In this paper the Authors report their 15 years experience in the laparoscopic surgical management of GERD. METHODS: From October 1991 to June 2005, 581 patients with GERD were treated by a laparoscopic fundoplication with a 360 degrees wrap (524 cases) or with a 270 degrees wrap (29 cases) or with a Collis gastroplasty (28 cases). RESULTS: The absence of intraoperative mortality, a low conversion to open surgery (0.7%) and a low postoperative morbidity (2.4%) with good clinical and instrumental long term results let the Authors to think that GERD represent an ideal indication for laparoscopic surgery. CONCLUSIONS: Good results are possible in the laparoscopic management of GERD with a precise selection of patients and with a good open and laparoscopic surgical experience in the management of functional esophageal diseases. PMID- 17122763 TI - [Technique, results and impact of induction chemotherapy in sleeve lobectomy for lung cancer]. AB - AIM: The aim of this study was to evaluate the safety of continuous nonabsorbable (3/0 polypropylene) sutures for sleeve lobectomy, and the influence of induction chemotherapy on postoperative outcome in patients with lung malignancies. METHODS: A review of a prospective database of a single surgeon identified 41 consecutive patients who underwent sleeve lobectomy from May 1998 to July 2003. Bronchial reconstruction was done placing two 3/0 polypropylene sutures at the far side of the cartilaginous wall and subsequently fixed. Afterwards, two running sutures were performed in order to obtain a telescopic anastomosis. RESULTS: Twenty-four patients (59%) underwent induction chemotherapy. There were 31 right upper, 3 left lower ''reverse'', and 7 left upper sleeve lobectomies with radical lymph node dissection. Eight patients underwent reconstruction of the pulmonary artery. There were 34 non-small cell lung cancers, 3 limited small cell lung cancers, 1 neuroendocrine large cell carcinoma, and 3 bronchial carcinoid tumors. N2, N1, and N0 diseases were found in 13, 12 and 16 patients, respectively. Post-operative morbidity and mortality were 14.5% (n=6) and 4.8% (n=2) (1 patient, 4%, after induction chemotherapy). The rate of postoperative anastomotic complications was 2.4% (n=1). Late bronchial stenosis developed in 3 cases, but all were successfully medically treated. Twenty-nine patients are still alive, 27 without evidence of disease. The overall 2-year probability of survival (Kaplan-Meier) was 59%. Induction chemotherapy did not influence postoperative morbidity/mortality (chi2 test: P=0.64/P=0.56). CONCLUSIONS: Continuous nonabsorbable suture for sleeve lobectomy is quick and technical easy to perform, with low postoperative morbidity/mortality; induction chemotherapy does not influence postoperative outcome in these patients. PMID- 17122764 TI - A dilemma: prophylaxis for pulmonary embolism after surgical or invasive interventions for hemodialysis. AB - AIM: This study was designed to investigate if it needs to do prophylaxis for pulmonary embolism in the patients treated with different kinds of dialysis or not, and if it is, to find a proper method for prophylaxis. METHODS: Ten numbers of patients with central venous catheters (CVC group), 13 numbers of patients with arteriovenous fistula (AVF group) were enrolled in this study. Eleven patients treated with peritoneal dialysis (PD group) were utilized as a control group. Clinical and laboratory examinations to exclude pulmonary embolism were carried out in both AVF and PD groups at the onset and after three months. Same examinations were performed in CVC group at the onset and after 3 weeks (mean: 21 days). Examinations to exclude pulmonary embolism consist of medical history, clinical examinations, d-dimer measures, chest x-ray, respiratory function tests, blood gas analyses, ventilation-perfusion scintigraphies. RESULTS: Neither clinical nor laboratory findings in any stages reflected any suspicion for pulmonary embolism. None of the patients in any groups was admitted with pulmonary embolism in any period of follow-up. There was not any statistically difference between the outcomes of all first examinations and of all second ones (P>0.005). Neither obvious nor subclinical pulmonary embolism was detected in any case. None of the patients had deep venous thrombosis in any stage of follow-up. CONCLUSIONS: Conventional techniques of haemodialysis do not lead to pulmonary embolism unless deep venous thrombosis due to any intervention occurs in the patients. Thus, prophylactic anticoagulant usage to prevent pulmonary embolism is not necessary in haemodialysis patients. To shorten the length of stay of central venous catheters is the most important factors for pulmonary embolism prophylaxis in haemodialysis patients. PMID- 17122765 TI - Emergency gastric ulcer complications in elderly. Factors affecting the morbidity and mortality in relation to therapeutic approaches. AB - AIM: In elderly the incidence of the emergency gastric ulcer complications, perforation and bleeding are increasing, with a difficult management of these patients for their concomitant diseases. The aim of this work is to analyze the therapeutical approach of emergency gastric ulcer complications in elderly patients, in order to establish the factors affecting the morbidity and mortality. METHODS: Patients older than 70 years, presenting gastric ulcer, observed in a tertiary University Hospital from 1995 to 2003, have been considered for the present study. Two groups of diseases have been examined: ulcer perforation and bleeding ulcer. Age, sex, risk factors, comorbidity, methods of diagnosis, ulcer characteristics, treatment, morbidity, mortality, hospitalization time and follow-up have been considered in each group. RESULTS: Thirteen elderly patients with perforated gastric ulcer have been observed: 9 (69.2%) females and 4 (30.8%) males with a mean age of 80.5 years (range 70-90). Four patients were hospitalized in suburban hospital with an average time between the diagnosis and the surgery of 36 h, while the remnants were hospitalized directly in our Department with a medium waiting time of about 2 h. The surgical procedures were: simple closure with omentum patch in 11 cases (84.6%), and antrectomy in 2 cases (15.4%), in which the antrum was multiply perforated. Two patients presented an ulcer larger than 2 cm treated with simple suture and omental patch without morbidity and mortality. Three patients (23%) died postoperatively, due to septic shock, ventricular fibrillation and intraoperative massive haemorrhage, 2 of these patients came from other hospitals. Twenty-eight elderly patients with bleeding gastric ulcer have been observed during the same period: 13 (46.4%) females and 15 (53.6%) males with a mean age of 79.6 years (range 71-91). Except 2 patients submitted to endoscopic treatment both with adrenaline injection, all the remnant patients were managed with medical therapy (H2-receptor antagonist or proton pump inhibitors and in 7 patients [24.1%] antihaemorrhage drugs), and clinical observation, with a endoscopic control 3-4 days after from the first endoscopy. One of the 2 patients endoscopically treated developed a ulcer perforation after 11 days, and the other one rebled, without possibility of any kind of treatment due to his instable condition of health. Three patients (10.7%) died during their hospital stay not for causes strictly due to the gastric haemorrhage. CONCLUSIONS: Our results suggest that the early diagnoses and early treatment are 2 basic factor on the prognosis of elderly patients with perforated gastric ulcer. The choice between simple closure, with or without vagotomy, or gastrectomy depends from preoperative and operative health conditions of the patient. In patients with ulcer larger than 2 cm, Graham's technique can be performed safely if the preoperative and intraoperative conditions are favourable. Elderly patients with gastric ulcer bleeding show an high risk of morbidity and mortality, related to the risk factors like non steroid anti-inflammatory drugs (NSAIDs) intake or smoke. Repeated endoscopy and antiulcer drugs can manage the high stage patients of Forrest's classification with a low rate of morbidity and mortality. According to literature surgical treatment should be reserved after the second failure of endoscopic treatment. PMID- 17122766 TI - Breast cancer surgery for the 21st century: the continuing evolution of minimally invasive treatments. AB - Since the 1950s, breast cancer surgery has been moving towards less invasive approaches for managing breast cancer, with sentinel lymph node biopsy (SLNB) and breast conservation therapy (BCT) now representing the standard of care for the majority of patients. Even as the use of SLNB is expanding to include patient groups that were previously thought to be poor candidates, questions remain about the optimal management of patients who are clinically node-negative but SLN positive, since more than half of these patients will prove to be pathologically node-negative. Various approaches are being developed to identify and treat those SLN-positive patients who are likely to have additional positive lymph nodes. The clinical significance of microscopic lesions in the SLN detected by immunohistochemistry continues to be debated--current standards recommend that isolated tumor cells (lesions no larger than 0.2 mm) be classified as pN0--but a definitive answer to this question awaits the completion of further studies. The unresolved questions about the best use of SLNB could become irrelevant with the ongoing development of new molecular prognostic indicators that may replace axillary lymph node status. Similarly, researchers are exploring ways of replacing BCT with ablation techniques that can remove the primary tumor without surgery. Although radiofrequency ablation, focused ultrasound, cryosurgery, and other approaches have captured the imagination of patients and clinicians alike, many technical difficulties remain. Among the most significant of these is the lack of truly precise imaging to locate tumors, estimate their true size, and follow treatment in real-time. These deficits may be filled by future developments in functional imaging (e.g., positron emission tomography) and nanobiotechnology. PMID- 17122767 TI - Extended redo sleeve resection for postoperative dehiscence after right upper bronchoplasty. AB - Complete anastomotic dehiscence after sleeve resection is a dramatic and often fatal event requiring an extremely aggressive and risky treatment; completion pneumonectomy represents the sole effective therapeutic option to rescue the patient, but postoperative mortality after this procedure is high. We report a case successfully treated by extended redo carinal sleeve resection after full bronchial dehiscence. This option should be taken into account in such a complication, mainly in patient with compromised respiratory function. PMID- 17122768 TI - [A surgical emergency: iatrogenic lesion of the spleen. Case report and review of the literature]. AB - The spleen is an organ often injured during surgical procedures. Iatrogenic lesions belong frequently to a low grade and can be treated with a conservative therapy. The surgeon may avoid the splenectomy by using new haemostatic agents as the patch of fibrinogen and thrombin in fixed combination. PMID- 17122769 TI - Systems biology: many things from one. PMID- 17122770 TI - Robustness-epistasis link shapes the fitness landscape of a randomly drifting protein. AB - The distribution of fitness effects of protein mutations is still unknown. Of particular interest is whether accumulating deleterious mutations interact, and how the resulting epistatic effects shape the protein's fitness landscape. Here we apply a model system in which bacterial fitness correlates with the enzymatic activity of TEM-1 beta-lactamase (antibiotic degradation). Subjecting TEM-1 to random mutational drift and purifying selection (to purge deleterious mutations) produced changes in its fitness landscape indicative of negative epistasis; that is, the combined deleterious effects of mutations were, on average, larger than expected from the multiplication of their individual effects. As observed in computational systems, negative epistasis was tightly associated with higher tolerance to mutations (robustness). Thus, under a low selection pressure, a large fraction of mutations was initially tolerated (high robustness), but as mutations accumulated, their fitness toll increased, resulting in the observed negative epistasis. These findings, supported by FoldX stability computations of the mutational effects, prompt a new model in which the mutational robustness (or neutrality) observed in proteins, and other biological systems, is due primarily to a stability margin, or threshold, that buffers the deleterious physico chemical effects of mutations on fitness. Threshold robustness is inherently epistatic-once the stability threshold is exhausted, the deleterious effects of mutations become fully pronounced, thereby making proteins far less robust than generally assumed. PMID- 17122771 TI - Identification and expansion of human colon-cancer-initiating cells. AB - Colon carcinoma is the second most common cause of death from cancer. The isolation and characterization of tumorigenic colon cancer cells may help to devise novel diagnostic and therapeutic procedures. Although there is increasing evidence that a rare population of undifferentiated cells is responsible for tumour formation and maintenance, this has not been explored for colorectal cancer. Here, we show that tumorigenic cells in colon cancer are included in the high-density CD133+ population, which accounts for about 2.5% of the tumour cells. Subcutaneous injection of colon cancer CD133+ cells readily reproduced the original tumour in immunodeficient mice, whereas CD133- cells did not form tumours. Such tumours were serially transplanted for several generations, in each of which we observed progressively faster tumour growth without significant phenotypic alterations. Unlike CD133- cells, CD133+ colon cancer cells grew exponentially for more than one year in vitro as undifferentiated tumour spheres in serum-free medium, maintaining the ability to engraft and reproduce the same morphological and antigenic pattern of the original tumour. We conclude that colorectal cancer is created and propagated by a small number of undifferentiated tumorigenic CD133+ cells, which should therefore be the target of future therapies. PMID- 17122772 TI - A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. AB - Colon cancer is one of the best-understood neoplasms from a genetic perspective, yet it remains the second most common cause of cancer-related death, indicating that some of its cancer cells are not eradicated by current therapies. What has yet to be established is whether every colon cancer cell possesses the potential to initiate and sustain tumour growth, or whether the tumour is hierarchically organized so that only a subset of cells--cancer stem cells--possess such potential. Here we use renal capsule transplantation in immunodeficient NOD/SCID mice to identify a human colon cancer-initiating cell (CC-IC). Purification experiments established that all CC-ICs were CD133+; the CD133- cells that comprised the majority of the tumour were unable to initiate tumour growth. We calculated by limiting dilution analysis that there was one CC-IC in 5.7 x 10(4) unfractionated tumour cells, whereas there was one CC-IC in 262 CD133+ cells, representing >200-fold enrichment. CC-ICs within the CD133+ population were able to maintain themselves as well as differentiate and re-establish tumour heterogeneity upon serial transplantation. The identification of colon cancer stem cells that are distinct from the bulk tumour cells provides strong support for the hierarchical organization of human colon cancer, and their existence suggests that for therapeutic strategies to be effective, they must target the cancer stem cells. PMID- 17122773 TI - Long-lasting arrest of murine polycystic kidney disease with CDK inhibitor roscovitine. AB - Polycystic kidney diseases (PKDs) are primarily characterized by the growth of fluid-filled cysts in renal tubules leading to end-stage renal disease. Mutations in the PKD1 or PKD2 genes lead to autosomal dominant PKD (ADPKD), a slowly developing adult form. Autosomal recessive polycystic kidney disease results from mutations in the PKHD1 gene, affects newborn infants and progresses very rapidly. No effective treatment is currently available for PKD. A previously unrecognized site of subcellular localization was recently discovered for all proteins known to be disrupted in PKD: primary cilia. Because ciliary functions seem to be involved in cell cycle regulation, disruption of proteins associated with cilia or centrioles may directly affect the cell cycle and proliferation, resulting in cystic disease. We therefore reasoned that the dysregulated cell cycle may be the most proximal cause of cystogenesis, and that intervention targeted at this point could provide significant therapeutic benefit for PKD. Here we show that treatment with the cyclin-dependent kinase (CDK) inhibitor (R)-roscovitine does indeed yield effective arrest of cystic disease in jck and cpk mouse models of PKD. Continuous daily administration of the drug is not required to achieve efficacy; pulse treatment provides a robust, long-lasting effect, indicating potential clinical benefits for a lifelong therapy. Molecular studies of the mechanism of action reveal effective cell-cycle arrest, transcriptional inhibition and attenuation of apoptosis. We found that roscovitine is active against cysts originating from different parts of the nephron, a desirable feature for the treatment of ADPKD, in which cysts form in multiple nephron segments. Our results indicate that inhibition of CDK is a new and effective approach to the treatment of PKD. PMID- 17122774 TI - Clustered DNA motifs mark X chromosomes for repression by a dosage compensation complex. AB - Gene expression in metazoans is regulated not only at the level of individual genes but also in a coordinated manner across large chromosomal domains (for example centromeres, telomeres and imprinted gene clusters) and along entire chromosomes (for example X-chromosome dosage compensation). The primary DNA sequence usually specifies the regulation of individual genes, but the nature of cis-acting information that controls genes over large regions has been elusive: higher-order DNA structure, specific histone modifications, subnuclear compartmentalization and primary DNA sequence are possibilities. One paradigm of chromosome-wide gene regulation is Caenorhabditis elegans dosage compensation in which a large dosage compensation complex (DCC) is targeted to both X chromosomes of hermaphrodites to repress transcript levels by half. This essential process equalizes X-linked gene expression between the sexes (XO males and XX hermaphrodites). Here we report the discovery and dissection of cis-acting sites that mark nematode X chromosomes as targets for gene repression by the DCC. These rex (recruitment element on X) sites are widely dispersed along X and reside in promoters, exons and intergenic regions. rex sites share at least two distinct motifs that act in combination to recruit the DCC. Mutating these motifs severely reduces or abolishes DCC binding in vivo, demonstrating the importance of primary DNA sequence in chromosome-wide regulation. Unexpectedly, the motifs are not enriched on X, but altering motif numbers within rex sites demonstrates that motif co-occurrence in unusually high densities is essential for optimal DCC recruitment. Thus, X-specific repression is established through sequences not specific to X. The distribution of common motifs provides the foundation for repression along an entire chromosome. PMID- 17122775 TI - p63 protects the female germ line during meiotic arrest. AB - Meiosis in the female germ line of mammals is distinguished by a prolonged arrest in prophase of meiosis I between homologous chromosome recombination and ovulation. How DNA damage is detected in these arrested oocytes is poorly understood, but it is variably thought to involve p53, a central tumour suppressor in mammals. While the function of p53 in monitoring the genome of somatic cells is clear, a consensus for the importance of p53 for germ line integrity has yet to emerge. Here we show that the p53 homologue p63 (refs 5, 6), and specifically the TAp63 isoform, is constitutively expressed in female germ cells during meiotic arrest and is essential in a process of DNA damage-induced oocyte death not involving p53. We also show that DNA damage induces both the phosphorylation of p63 and its binding to p53 cognate DNA sites and that these events are linked to oocyte death. Our data support a model whereby p63 is the primordial member of the p53 family and acts in a conserved process of monitoring the integrity of the female germ line, whereas the functions of p53 are restricted to vertebrate somatic cells for tumour suppression. These findings have implications for understanding female germ line fidelity, the regulation of fertility and the evolution of tumour suppressor mechanisms. PMID- 17122776 TI - Variability and memory of protein levels in human cells. AB - Protein expression is a stochastic process that leads to phenotypic variation among cells. The cell-cell distribution of protein levels in microorganisms has been well characterized but little is known about such variability in human cells. Here, we studied the variability of protein levels in human cells, as well as the temporal dynamics of this variability, and addressed whether cells with higher than average protein levels eventually have lower than average levels, and if so, over what timescale does this mixing occur. We measured fluctuations over time in the levels of 20 endogenous proteins in living human cells, tagged by the gene for yellow fluorescent protein at their chromosomal loci. We found variability with a standard deviation that ranged, for different proteins, from about 15% to 30% of the mean. Mixing between high and low levels occurred for all proteins, but the mixing time was longer than two cell generations (more than 40 h) for many proteins. We also tagged pairs of proteins with two colours, and found that the levels of proteins in the same biological pathway were far more correlated than those of proteins in different pathways. The persistent memory for protein levels that we found might underlie individuality in cell behaviour and could set a timescale needed for signals to affect fully every member of a cell population. PMID- 17122777 TI - How Neanderthal molar teeth grew. AB - Growth and development are both fundamental components of demographic structure and life history strategy. Together with information about developmental timing they ultimately contribute to a better understanding of Neanderthal extinction. Primate molar tooth development tracks the pace of life history evolution most closely, and tooth histology reveals a record of birth as well as the timing of crown and root growth. High-resolution micro-computed tomography now allows us to image complex structures and uncover subtle differences in adult tooth morphology that are determined early in embryonic development. Here we show that the timing of molar crown and root completion in Neanderthals matches those known for modern humans but that a more complex enamel-dentine junction morphology and a late peak in root extension rate sets them apart. Previous predictions about Neanderthal growth, based only on anterior tooth surfaces, were necessarily speculative. These data are the first on internal molar microstructure; they firmly place key Neanderthal life history variables within those known for modern humans. PMID- 17122778 TI - A bacterial dynamin-like protein. AB - Dynamins form a superfamily of large mechano-chemical GTPases that includes the classical dynamins and dynamin-like proteins (DLPs). They are found throughout the Eukarya, functioning in core cellular processes such as endocytosis and organelle division. Many bacteria are predicted by sequence to possess large GTPases with the same multidomain architecture that is found in DLPs. Mechanistic dissection of dynamin family members has been impeded by a lack of high resolution structural data currently restricted to the GTPase and pleckstrin homology domains, and the dynamin-related human guanylate-binding protein. Here we present the crystal structure of a cyanobacterial DLP in both nucleotide-free and GDP-associated conformation. The bacterial DLP shows dynamin-like qualities, such as helical self-assembly and tubulation of a lipid bilayer. In vivo, it localizes to the membrane in a manner reminiscent of FZL, a chloroplast-specific dynamin-related protein with which it shares sequence similarity. Our results provide structural and mechanistic insight that may be relevant across the dynamin superfamily. Concurrently, we show compelling similarity between a cyanobacterial and chloroplast DLP that, given the endosymbiotic ancestry of chloroplasts, questions the evolutionary origins of dynamins. PMID- 17122779 TI - Slc11a1 (formerly NRAMP1) gene modulates both acute inflammatory reactions and pristane-induced arthritis in mice. AB - Mice selected for the maximum acute inflammatory reaction (AIRmax) are highly susceptible to pristane-induced arthritis (PIA), whereas mice selected for the minimum response (AIRmin) are resistant. These lines show distinct patterns of leukocyte infiltration and R and S allele frequency disequilibrium of the solute carrier family 11a member 1 (Slc11a1) gene. In order to study the interactions of the Slc11a1 R and S alleles with the inflammation modulating Quantitative Trait Loci (QTL) during PIA development, homozygous AIRmax(RR), AIRmax(SS), AIRmin(RR) and AIRmin(SS) lines were produced by genotype-assisted breedings. These mice received two intraperitoneal injections of 0.5 ml pristane at 60-day intervals, and the subsequent development of arthritis was assessed for 210 days. Cytokine secreting cell profiles were investigated using enzyme-linked immunospot. Arthritis incidence in AIRmax(RR) mice reached 29%, whereas PIA incidence in AIRmax(SS) mice was 70% by day 180. AIRmin(RR) mice were resistant, whereas 13.3% of AIRmin(SS) mice became arthritic. The presence of the defective S allele also increased arthritis severity, although acute inflammation was higher in mice bearing the R allele. A predominant Th0/Th2-type response in Slc11a1(SS) mice was observed. These results indicate that Slc11a1 is a strong candidate for the QTL modulating acute inflammation and for PIA. PMID- 17122780 TI - Genome-wide scan for visceral leishmaniasis susceptibility genes in Brazil. AB - A genome-wide scan was conducted for visceral leishmaniasis (VL) in Brazil. Initially, 405 markers were typed in 22 multicase pedigrees (28 nuclear families; 174 individuals; 66 affected). Non-parametric multipoint analysis detected nine chromosomal regions with provisional evidence (logarithm of the odds (LOD) scores 0.95-1.66; 0.003 2)-beta-D-GalpA-(1--> 3)-alpha-D-GalpNAc-(1--> 4) alpha-D-GalpA-(1--> 3)-beta-D-GlcpNAc-(1--> The same structure has been reported previously for the O-polysaccharides of P. mirabilis CCUG 10701 (O74) and P. mirabilis 41/57 (O23), except that they contain O-acetyl groups in non stoichiometric quantities. Serological studies showed the antigenic identity of the three strains and their close serological relatedness to P. vulgaris 44/57. CONCLUSIONS: Based on the OPS structures and serological data, it is suggested to classify P. mirabilis 41/57, TG 115, and CCUG 10701 into one subgroup and P. mirabilis 42/57 and P. vulgaris 43/57 and 44/57 into another subgroup of the Proteus O23 serogroup. PMID- 17122882 TI - Control of immune responses by immunoregulatory T cells. AB - Immunoregulatory T cells play a key role in modifying the immune responses to self antigens, tumor antigens, and pathogenic organisms. This review summarizes recent data on naturally occurring CD4(+) regulatory T cells that constitutively express CD25 (CD25(+)T(reg)). We examine the markers that can be used to differentiate these cells from effector T cells, what is known about their mode of action in controlling the activity of effector T cells, the antigenic specificity of CD25(+)T(reg), and their ability to survive and to be selected in vivo. We also summarize specific information on the role of CD25(+)T(reg) in controlling anti-tumor responses, an area were manipulation of this subset holds particular clinical promise. PMID- 17122883 TI - Hematopoietic cell transplantation for chronic myeloproliferative disorders. AB - Myeloproliferative disorders, including chronic idiopathic myelofibrosis (CIMF), polycythemia vera (PV), essential thrombocythemia (ET), and chronic myelomonocytic leukemia (CMML), are clonal diseases of hematopoietic stem or precursor cells. They often show a protracted or chronic course; however, all have the potential of progressing to severe marrow failure, associated with myelofibrosis, or of transforming into acute leukemia. At that point, hematopoietic cell transplantation (HCT) is the only current treatment strategy with curative potential. If transplantation is being considered and a suitable donor is available, HCT should be carried out before leukemic transformation has occurred, as the success rate of HCT declines steeply in patients who have evolved to leukemia. As many as 75-80% of patients with the original diagnoses of PV or ET, about 65-70% with CIMF, and 45% of patients with CMML are surviving long term after allogeneic HCT using conventional transplant regimens, with follow-up now extending to 15 years. Results with HLA-identical related and unrelated donors are comparable. Major risk factors for the outcome after HCT are the disease stage, the presence of comorbid conditions, and patient age. The development of reduced-intensity conditioning regimens has allowed for successful HCT even for older patients and patients with comorbid conditions. Studies on disease mechanisms, including the recent characterization of an activating mutation in JAK2, may provide additional prognostic guidance and are likely to lead to the development of novel treatment strategies, which will require continuous reassessment as to the optimum timing of HCT. PMID- 17122884 TI - Modulation of the immune response by extracellular matrix proteins. AB - Inflammation entrains a focused and coordinated response from many different elements. Soluble factors such as chemokines and cytokines direct the recruitment, differentiation, and fate of leukocytes. Cells and pathogens are killed and consumed, yet where the response is effective, inflammation will melt away, leaving a healthy functioning tissue. All this commonly takes place in an environment known as the extracellular matrix (ECM). The ECM is not a passive partner in the process and recent work demonstrates the important role that proteins found in this environment play in connecting different parts of the immune response together. In this review we will focus on these connections and the proteins that make them. One emerging trend that we will highlight is the ability of endogenous molecules to interact with receptors that are better known as sensors of the molecular fingerprints of infection. We propose that this may be particularly relevant in the context of autoimmunity, since the provision of such signals may be crucial in breaking tolerance. PMID- 17122885 TI - Gunther Tulip retrievable inferior vena caval filters: indications, efficacy, retrieval, and complications. AB - PURPOSE: We evaluated the Gunther Tulip (GT) retrievable inferior vena cava (IVC) filter with regard to indications, filtration efficacy, complications, retrieval window, and use of anticoagulation. METHOD: A retrospective study was performed of 147 patients (64 men, 83 women; mean age 58.8 years) who underwent retrievable GT filter insertion between 2001 and 2005. The indications for placement included a diagnosis of pulmonary embolism or deep venous thrombosis with a contraindication to anticoagulation (n = 68), pulmonary embolism or deep venous thrombosis while on anticoagulation (n = 49), prophylactic filter placement for high-risk surgical patients with a past history of pulmonary embolism or deep venous thrombosis (n = 20), and a high risk of pulmonary embolism or deep venous thrombosis (n = 10). Forty-nine of the 147 patients did not receive anticoagulation (33.7%) while 96 of 147 patients did, 82 of these receiving warfarin (56.5%), 11 receiving low-molecular weight heparins (7.58%), and 3 receiving antiplatelet agents alone (2.06%). RESULTS: Filter placement was successful in 147 patients (100%). Two patients had two filters inserted. Of the 147 patients, filter deployment was on a permanent basis in 102 and with an intention to retrieve in 45 patients. There were 36 (80%) successful retrievals and 9 (20%) failed retrievals. The mean time to retrieval was 33.6 days. The reasons for failed retrieval included filter struts tightly adherent to the IVC wall (5/9), extreme filter tilt (2/9), and extensive filter thrombus (2/9). Complications included pneumothorax (n = 4), failure of filter expansion (n = 1), and breakthrough pulmonary embolism (n = 1). No IVC thrombotic episodes were recorded. DISCUSSION: The Gunther Tulip retrievable filter can be used as a permanent or a retrievable filter. It is safe and efficacious. GT filters can be safely retrieved at a mean time interval of 33.6 days. The newly developed Celect filter may extend the retrieval interval. PMID- 17122886 TI - Long-term results after placement of aortic bifurcation self-expanding stents: 10 year mortality, stent restenosis, and distal disease progression. AB - PURPOSE: To retrospectively evaluate the 10 year follow-up results in patients who had "kissing" self-expanding stent aortic bifurcation reconstruction. METHODS: Forty-three patients were treated with "kissing" self-expanding stents for aortoiliac occlusive disease. Early follow-up with clinical and ankle brachial pressure indices (ABPI) was performed at 3, 6, 12, and 24 months and with intra-arterial digital subtraction angiography at 12-24 months; clinical and angiographic follow-up was performed for symptom recurrence up to 10 years after treatment. Retrospective record review was performed to assess mortality, clinical patency, angiographic patency, and secondary assisted patency of both stents and downstream peripheral vessels at 5 and 10 years follow-up. RESULTS: The 2 year primary angiographic and secondary assisted stent patencies were 89% and 93%, respectively. At 10 years follow-up in 40 patients the mortality was 38% (due to myocardial infarction, stroke, chronic renal failure, malignancy, and liver failure). At 5 and 10 years follow-up the primary clinical stent patency was 82% and 68%, and the secondary assisted stent patency 93% and 86%, respectively. At 5 and 10 years, the distal vessel patency was 86% and 72%, and the secondary assisted distal vessel patency treated by surgical or endovascular techniques was 94% and 88%, respectively. At 10 years there was no limb loss. CONCLUSION: The long-term (10 year) results of aortic bifurcation arterial self expanding stent placement in patients with arterial occlusive disease show a 10 year primary stent patency rate of 68% but a secondary assisted patency rate of 86%. In addition there is a high overall mortality due to other cardiovascular causes and the rate of distal disease progression and loss of patency is similar to the loss of stent patency rate. PMID- 17122887 TI - Percutaneous recanalization of acute internal carotid artery occlusions in patients with severe stroke. AB - BACKGROUND: Sudden symptomatic occlusions of the proximal internal carotid artery (ICA) resulting in severe middle cerebral artery (MCA) ischemia and stroke are usually not accessible by rt-PA thrombolysis and the prognosis is usually very poor. Mechanical recanalization of the proximal ICA combined with intravenous and intra-arterial thrombolysis was therefore used as a rescue procedure. METHODS: Ten patients (9 men, 1 woman; mean age 56.1 years) were treated with emergency recanalization of the proximal carotid artery by using stents and/or balloon angioplasty as a rescue procedure. Three patients showed dissection, and 7 had atherothrombotic occlusions. Nine of 10 presented with an initial modified Rankin Scale (mRS) of 5, the remaining patient with mRS 4 (average NIHSS 21.4). After sonographic confirmation of ICA with associated MCA/distal ICA occlusion and bridging with rt-PA (without abciximab) an emergency angiography was performed with subsequent mechanical recanalization by percutaneous transluminal angioplasty (PTA) (n = 1) or primary stenting (n = 9) using self-expanding stents. Distal protection was used in 1 of 10 patients. RESULTS: Recanalization of the proximal ICA was achieved in all. At least partial recanalization of the intracerebral arteries was achieved in all, and complete recanalization in 5. In 4 of 10 patients limited hemorrhage was detected during CT controls. Major complications included 2 patients who had to undergo hemicraniectomy. One patient died from malignant infarction. At the time of discharge from the stroke unit 9 of 10 patients had improved markedly, 5 patients having an mRS of < or =2, and 3 patients a mRS of 3. At control after a mean of 20 weeks, 7 of 8 (88%) patients had a mRS < or =2, and 1 a mRS of 3. CONCLUSIONS: Primary mechanical recanalization of ICA occlusions by stent and PTA combined with fibrinolysis and/or GPIIb/IIIa-receptor antagonists seems to be feasible to improve patient outcome significantly. PMID- 17122888 TI - Treatment of secondary stent-graft collapse after endovascular stent-grafting for iliac artery pseudoaneurysms. AB - We report the case of a patient who developed an asymptomatic pseudoaneurysm in the left external iliac artery after transplant nephrectomy. The pseudoaneurysm most probably arose as a suture aneurysm from the external iliac artery after removal of the graft renal artery. Obviously we can not exclude the possibility it was a true aneurysm, although this seems much less likely. The pseudoaneurysm was detected during a routine CT scan and was treated interventionally with a stent-graft. One month later the asymptomatic patient underwent a vascular ultrasound examination including color Doppler, power Doppler, and B-flow as a routine control. An endoleak with collapse of the stent-graft was diagnosed. There was no evidence of stent infection. At a reintervention, the pseudoaneurysm was successfully treated using two uncovered Palmaz stents at the proximal and distal edge of the stent graft. Peri- and post-interventional ultrasound and CT angiography confirmed the exclusion of the aneurysm without an endoleak. PMID- 17122889 TI - Interstitial remodeling in beta1-adrenergic receptor transgenic mice. AB - BACKGROUND: Inhibition of proteolytic MMP activity could be a therapeutic approach to prevent ventricular dilatation by diminishing collagen matrix turnover and interstitial fibrosis. We investigated the time-course of MMP/TIMP activity during transition from hypertrophy to ventricular dilatation in transgenic mice with myocyte overexpression of the human beta1-adrenergic receptor (beta1TG). These beta1TG mice were studied at 3 (normal function), 5 (hypertrophy) and 12 (ventricular dilatation) months of age compared to age matched controls (WT). METHODS: Picro Sirius red staining and real-time PCR were performed for total collagen and for collagen type I and III quantification, respectively. MMP-activity assays (zymography), immunoblotting and real-time PCR experiments were done for gelatinase- (MMP-2, -9), collagenase- (MMP-1, -13), membrane-type MMP- (MT1- MMP; MMP-14) and TIMP expression measurements. To investigate beta1-integrin activity, integrin-linked kinase (ILK) expression was measured by immunoblotting. RESULTS: Compared to WT with normal cardiac function, interstitial collagen type I and III mRNA and protein expression increased 3.6 fold in beta1TG at 5 months of age with moderate fibrosis and cardiomyocyte hypertrophy and 17-fold in beta1TG at 12 months of age with severe fibrosis and ventricular dilatation. Protein expression of the collagenases MMP-1 and -13 as well as the gelatinase proMMP-2 increased in the beta1TG group with cardiac hypertrophy. Maximal activity of the gelatinase MMP-2 (3.5-fold vs.WT) was measured in beta1TG at 12 months of age with severe fibrosis and ventricular dilatation, accompanied by coexpression of MT1- MMP (3.8-fold vs.WT) colocalized to the cell membranes. CONCLUSION: These data provide evidence that sympathetic overactivation can trigger interstitial matrix remodeling and fibrosis by induction of MMP/TIMP activity. In particular gelatinolytic MMP-2 activity accompanies ventricular dilatation and the development of heart failure. PMID- 17122890 TI - Maintaining serum response factor activity in the older heart equal to that of the young adult is associated with better cardiac response to isoproterenol stress. AB - To understand the effect of transcription regulation in modulating cardiac aging, we sought to study the role of serum response factor (SRF), a key transcription factor in the heart that is normally increased with senescence and also in congestive heart failure. A Tet-Off gene expression system was used for cardiac specific over-expression of a mutant SRF protein. In these binary transgenic mice, there is no age-related increase in SRF protein expression; in fact, there appeared to be a mild reduction of SRF protein (Mild-R SRF Tg). The older, middle aged (15 mo) Mild-R SRF Tg mice appeared healthier and were better able to maintain their left ventricular systolic pressure (LVSP) in response to moderate a-adrenergic stimulation compared with age-matched Non-Tg mice, which demonstrated a negative ionotropic response. The Mild-R SRF Tg hearts had lower mRNA expression of BNP (p < 0.05), and the sodium calcium exchanger (p < 0.05), compared to Non-Tg. Mild-R SRF Tg had higher mRNA levels of SERCA2 (p < 0.05) and ryanodine receptor 2 (p < 0.05) compared to Non-Tg hearts. These findings suggest that preventing the age-associated increase in SRF is associated with better preserved intracellular calcium handling and functional response to stress; it might be advantageous for the older adult heart. This mouse model could be helpful in elucidating the molecular mechanisms underlying certain age-related changes in cardiac reserve capacity and response to stress. PMID- 17122891 TI - Enhanced glutathione S-transferase expression in 2-hydroxyamino-1-methyl-6 phenylimidazo[4,5-b]pyridine-resistant IEC-18 cells. AB - In the present study we show that repeated exposure of the rat intestinal epithelial cell line IEC-18 to 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5 b]pyridine (N-OH-PhIP), from a toxicological point of view the most relevant phase-1 metabolite of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP, the main heterocyclic aromatic amine present in processed meat), led to the selection of N-OH-PhIP-resistant IEC-18 cells. This phenomenon was accompanied by a fivefold increase in total glutathione S-transferase (GST) activity, measured with the broad-spectrum substrate 1-chloro-2,4-dinitrobenzene, in the N-OH-PhIP resistant IEC-18 cells. Furthermore, a Western blotting analysis revealed that the expression of GST subunits A1, A3, A4, M1 and P1 was enhanced in the N-OH PhIP-resistant IEC-18 cells. PMID- 17122892 TI - High production of laccase by a new basidiomycete, Trametes sp. AB - A new basidiomycete, Trametes sp. 420, produced laccase at 6,810 U l(-1) (268 mg, 25.4 U mg(-1) protein for guaiacol) in glucose medium and 7,870 U l(-1) (310 mg) in cellobiose medium with induction by 0.5 mM Cu(2+) and 6 mM o-toluidine. Laccase isozyme E (LacE) was the sole laccase in the fermentation products. It was stable at pH 5-9 and below 70 degrees C over 30 min. The K (m) values of LacE for four substrates (guaiacol ABTS, 2,6-dimethoxyphenol and syringaldazine) varied from 5 to 245 muM. The activity of LacE was strongly inhibited by NaN(3) but not by EDTA or dimethylsulfoxide. LacE at 0.5 U l(-1) could decolorize industrial dyes. The open reading frame of the lacE gene was 2,130 bp and was interrupted by 10 introns. It displayed a high homology to laccases from other fungi. PMID- 17122893 TI - Transcription mapping and characterization of proteins produced from early region 4 of porcine adenovirus type 3. AB - The early region 4 (E4) of porcine adenovirus 3 (PAdV-3) was characterized by Northern blot, rapid amplification of cDNA ends (RACE), RT-PCR and cDNA sequence analysis. Northern blot analysis revealed three different classes of transcripts, which appeared and peaked at different times post-infection. The RT-PCR, RACE and cDNA sequence analysis identified nine major E4 transcripts, all of which shared a 107-bp 5' leader sequence and a 126-bp 3' terminus. These transcripts have one to three introns removed. Interestingly, of the nine major transcripts, there was one fusion transcript of ORFp1 and ORFp7 (ORFp1/7), which codes for a protein of 119 amino acids. All transcripts initiated at nucleotide 33740 of the PAdV-3 genome. To identify proteins, rabbit antiserum was prepared using a bacterial fusion protein encoding p2, p3, p4 or p7 proteins. Serum against p2, p3 and p4 immunoprecipitated proteins of 13.5, 13.6 and 15.3 kDa, respectively, in in-vitro transcribed and translated mRNA and in PAdV-3-infected cells. Serum against p7 immunoprecipitated a protein of 19.8 kDa in in-vitro transcription and translation analysis but recognized two proteins of 19.8 kDa (encoded by ORFp7) and 14 kDa (encoded by the fusion transcript ORF1/7) in PAdV-3-infected cells. The protein encoded by ORFp2 was localized in the nucleus of PAdV-3-infected cells. The proteins encoded by ORFp3 and ORFp7?ORFp1/7 were detected in the cytoplasm of PAdV-3-infected cells. However, the protein encoded by ORFp4 was observed both in the cytoplasm and nucleus of PAdV-3-infected cells. PMID- 17122894 TI - Characterisation of several heterogeneous species of defective RNAs derived from RNA 3 of cucumber mosaic virus. AB - Preparations of double-stranded RNAs (dsRNAs) extracted from Nicotiana tabacum cv Xanthi plants infected with a subgroup IB isolate of Cucumber mosaic virus (CMV) were found to contain a heterogeneous population of defective RNAs (D-RNAs) derived from RNA 3. Characterised D-RNAs ranged in size from 1.5 to 1.9 kb and were derived either by a single in-frame deletion within the 3a or 3b genes or by means of double in-frame deletions within both genes. Also, northern blot hybridisation showed two other types of RNA derived from RNA 3: (a) RNA species of ca. 0.7 kb containing the 3'-terminus but lacking the 5'-terminus, which could be 3'-coterminal subgenomic of D-RNAs derived from the 3b gene and (b) RNA species of unknown origin of ca. 0.8 kb containing the 5'-terminus but lacking the 3'-terminus. PMID- 17122895 TI - The major population of PHA-stimulated PBMC infected by R5 or X4 HIV variants after a single cycle of infection is predominantly composed of CD45RO+CD4+ T lymphocytes. AB - PHA-stimulated peripheral blood mononuclear cells (PBMCs) are widely used for investigating replication and neutralization of HIV primary isolates in vitro. The objective of this study was to identify the T lymphocyte subset(s) that are found infected after one replication cycle by either R5- or X4-HIV-1 variants in PHA-stimulated PBMCs from healthy donors. Infected T lymphocytes were detected by intracellular p24 staining and characterized by cell surface immunophenotyping using flow cytometry. The predominant lymphocyte subset expressing p24 after 24 h of infection with either R5 or X4 HIV-1 strains was found to exhibit mainly the memory CD45RO phenotype, a greater percentage of CD62L(+)CD45RO(+) central memory T lymphocytes was infected with X4 HIV strains. Although some CD45RA(+) lymphocytes were also infected, these cells co-expressed CD45RO(+). The proportion of lymphocytes expressing CD4 and CD4/CD45RO decreased by 20% after 24 h of infection. A 2-fold decrease of CD4(+)CD8(+) T lymphocytes could also be recorded, even though this subset accounted for less than 5% of total lymphocytes in control cultures. Moreover, CD4(+)CD8(+) T cells further decreased by 90% after 4 days of infection, a time at which they scored p24(+). Therefore, our results indicate that the in vitro infection system of PHA-stimulated PBMC utilized in neutralization assays provides an appropriate model for the study of infected CD45RO(+) lymphocytes but not CD45RA(+) lymphocytes. PMID- 17122896 TI - Primary mouse dermal fibroblast cell cultures as an in vitro model system for the differential pathogenicity of cross-species herpesvirus papio 2 infections. AB - Infection of mice with herpesvirus papio 2 (HVP2) parallels zoonotic monkey B virus infections. A major benefit of the HVP2/mouse model is the existence of two HVP2 subtypes: HVP2nv rapidly invades and destroys the CNS while HVP2ap produces no clinical signs and mild histopathological lesions. However, in the natural baboon host, no difference in pathogenicity of HVP2 subtypes is evident. Primary dermal fibroblast cells were evaluated as a model system for defining virus-host interactions that influence the outcome of a cross-species infection. No differences in plaque formation or virus replication were observed between HVP2 subtypes in primary baboon dermal fibroblast cultures. In contrast, when primary mouse dermal fibroblasts (PMDF) were infected, HVP2nv replicated to higher titers and was more efficient at shutting down host-cell protein synthesis compared to HVP2ap. HVP2ap-infected PMDF cells produced more IFN-beta compared to HVP2nv, and IFN-beta pretreatment of PMDF cultures inhibited HVP2ap replication but did not affect HVP2nv. The differential pathogenicity of HVP2 subtypes in mice and the lack of such differences in the natural baboon host are recapitulated in the primary dermal fibroblast cell culture system. This model may prove useful in examining early, local, host responses that influence the outcome of cross species infections. PMID- 17122897 TI - Sequence analysis of the complete genomes of two Rice tungro spherical virus isolates from India. PMID- 17122898 TI - [Pediatric respiratory tract stenoses: are subspecialization and the development of specialist centers necessary?]. PMID- 17122901 TI - Isolation and characterization of Ty1/copia-like retrotransposons in mung bean (Vigna radiata). AB - Two Ty1/copia-like retrotransposons, RTvr1 and RTvr2, were isolated from mung bean (Vigna radiata (L.) Wilczek) genomic DNA and are the first complete elements of this kind to be reported in this legume. Nucleotide sequence analyses revealed that both elements are AT-rich (60% and 61%, respectively) and are flanked by a target-site duplication of 5 bp. The structures of RTvr1 and RTvr2 are those of typical long terminal repeat retrotransposons. Both transposons were able to produce putative proteins with the domain order of Gag-protease-integrase-reverse transcriptase-RNase H, indicating that RTvr1 and RTvr2 belong to the Ty1/copia like retrotransposons. Except for a 2,500-bp insertion region in RTvr2, the overall similarity between RTvr1 and RTvr2 is 92%. Dot blots showed that these two retroelements were present at a copy number of 120 per mung bean haploid genome. Multiple sequence alignments showed that the conserved motifs of the aspartic proteases, integrase, reverse transcriptase, and the RNase H in the Ty1/copia-like group all exist in RTvr1 and RTvr2. PMID- 17122900 TI - [Pharmacological relapse prevention in alcohol and tobacco dependence]. AB - A better understanding of the neurobiological substrates of alcohol and tobacco dependence has been accompanied by the increasing role of pharmacological relapse prevention. In alcohol dependence, substances which are able to block or ameliorate alcohol craving improve the maintenance of abstinence in combination with psychosocial treatment. In particular, the N-methyl-D-aspartate receptor antagonist acamprosate and the opiate receptor antagonist naltrexone have been shown to be effective in numerous trials. Administration starts immediately after detoxification and should last for 12 months. This treatment, however, is not a replacement for the psychosocial treatment of the alcohol dependence. Compared with placebo treatment, approximately twice as many patients remain abstinent under acamprosate 1 year after the end of treatment. The most widely studied and used pharmacotherapy for the treatment of tobacco dependence is nicotine containing medications. Nicotine replacement therapies (patch, gum, tablet, inhaler) have been shown to reduce tobacco craving and enhance abstinence. The only non-nicotine medication that has been approved in Germany is the antidepressant bupropion. Current studies identify particularly suitable subgroups of patients responding to more individualized pharmacological relapse prevention. A specific application of these substances will optimise the outcome for alcohol and tobacco dependent patients. PMID- 17122902 TI - Severe anxiety symptomatology and HIV risk behavior among Hispanic injection drug users in Puerto Rico. AB - Despite an overall decrease in AIDS incidence in Puerto Rico, our studies continue to show high prevalence of HIV risk behaviors among injection drug users (IDUs). This study seeks to evaluate whether the occurrence of injection-related and sex-related HIV risk behaviors among IDUs in Puerto Rico varies with the presence of anxiety symptomatology. Subjects included 557 IDUs, recruited from street settings in poor neighborhoods in Puerto Rico. Symptoms of severe anxiety were reported by 37.1% of the study sample. Participants with severe anxiety symptoms were more likely to share needles, cotton, and rinse water; to pool money to buy drugs; and to engage in backloading, than those without severe anxiety symptoms. Participants with severe anxiety symptomatology were also more likely to practice unprotected vaginal or oral sex. The findings from this study alert HIV prevention and treatment programs to the need to address anxiety disorders within their programs. PMID- 17122903 TI - Accessory cardiac bronchus and tracheal bronchus anomalies: CT-bronchoscopy and CT-bronchography findings. AB - Most common developmental anomalies of bronchial tree include accessory cardiac bronchus (ACB) and tracheal bronchus (TB). Minor bronchial anomalies include variants of TB, displaced segmental bronchi, and bronchial agenesis. We present CT-bronchoscopy and CT-bronchography findings of three cases with either ACB or TB. Recognition of these anomalies is important, as associated clinical complications, including recurrent episodes of infection, hemoptysis, and perhaps malignancies may be anticipated in a small percentage of patients. PMID- 17122904 TI - Effects of levosimendan on myocardial infarct size and hemodynamics in a closed chest porcine ischemia-reperfusion model. AB - INTRODUCTION: Levosimendan is a positive inotropic drug with vasodilator action and proposed myocardioprotective properties. In a canine model, levosimendan increased coronary collateral flow and reduced myocardial infarct size (IS). We investigated the effect of levosimendan on IS and hemodynamics in the closed chest porcine ischemia-reperfusion model, which is devoid of coronary collaterals. METHODS: Infusion with levosimendan (0.2 microg/kg/min following a bolus of 24 microg/kg) or saline was initiated 30 min prior to ischemia in anaesthetized pigs (n = 10 in both groups). Balloon occlusion of the left anterior descending coronary artery for 45 min was followed by 2 1/2 h of reperfusion. Hemodynamics were monitored with a Swan-Ganz catheter and a left ventricular pressure micromanometer. Left ventricular systolic and diastolic function was estimated by dP/dt(max) and tau, respectively. Myocardial area at risk (AAR) and IS were assessed in vivo by myocardial perfusion imaging (MPI) and ex vivo by histopathology (fluorescein staining for AAR, tetrazolium staining for IS). RESULTS: Prior to ischemia, levosimendan improved left ventricular systolic and diastolic function with coincident preload and afterload reduction. Cardiac output increased by 10 +/- 4% (p = 0.04), dP/dt(max) by 15 +/- 5% (p = 0.01). Pulmonary capillary wedge pressure decreased by 18 +/- 3% (p = 0.04), tau by 11 +/- 2% (p = 0.001), and mean arterial pressure by 11 +/- 2% (p < 0.001). A similar trend was observed during ischemia-reperfusion. The ratio of IS/AAR was not reduced by levosimendan compared to saline as evaluated by histopathology (76 +/- 4% vs. 64 +/- 7%, p = 0.12) and by MPI (94 +/- 2% vs. 87 +/- 5%, p = 0.14). CONCLUSION: Levosimendan improves hemodynamics but does not reduce IS in an ischemia-reperfusion model without coronary collaterals. PMID- 17122905 TI - A forecast of the future for biomaterials. PMID- 17122906 TI - A tribute to Professor Larry Hench. PMID- 17122907 TI - The story of Bioglass. AB - Historically the function of biomaterials has been to replace diseased or damaged tissues. First generation biomaterials were selected to be as bio-inert as possible and thereby minimize formation of scar tissue at the interface with host tissues. Bioactive glasses were discovered in 1969 and provided for the first time an alternative; second generation, interfacial bonding of an implant with host tissues. Tissue regeneration and repair using the gene activation properties of Bioglass provide a third generation of biomaterials. This article reviews the 40 year history of the development of bioactive glasses, with emphasis on the first composition, 45S5 Bioglass, that has been in clinical use since 1985. The steps of discovery, characterization, in vivo and in vitro evaluation, clinical studies and product development are summarized along with the technology transfer processes. PMID- 17122908 TI - The surface functionalization of 45S5 Bioglass-based glass-ceramic scaffolds and its impact on bioactivity. AB - The first and foremost function of a tissue engineering scaffold is its role as a substrate for cell attachment, and their subsequent growth and proliferation. However, cells do not attach directly to the culture substrate; rather they bind to proteins that are adsorbed to the scaffold's surface. Like standard tissue culture plates, tissue engineering scaffolds can be chemically treated to couple proteins without losing the conformational functionality; a process called surface functionalization. In this work, novel highly porous 45S5 Bioglass-based scaffolds have been functionalized applying 3-AminoPropyl-TriethoxySilane (APTS) and glutaraldehyde (GA) without the use of organic solvents. The efficiency and stability of the surface modification was assessed by X-ray photoemission spectroscopy (XPS). The bioactivity of the functionalized scaffolds was investigated using simulated body fluid (SBF) and characterized by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and X-ray diffraction (XRD). It was found that the aqueous heat-treatment applied at 80 degrees C for 4 hrs during the surface functionalization procedure accelerated the structural transition of the crystalline Na2Ca2Si3O9 phase, present in the original scaffold structure as a result of the sintering process used for fabrication, to an amorphous phase during SBF immersion. The surface functionalized scaffolds exhibited an accelerated crystalline hydroxyapatite layer formation upon immersion in SBF caused by ion leaching and the increased surface roughness induced during the heat treatment step. The possible mechanisms behind this phenomenon are discussed. PMID- 17122909 TI - Controlling ion release from bioactive glass foam scaffolds with antibacterial properties. AB - Bioactive glass scaffolds have been produced, which meet many of the criteria for an ideal scaffold for bone tissue engineering applications, by foaming sol-gel derived bioactive glasses. The scaffolds have a hierarchical pore structure that is very similar to that of cancellous bone. The degradation products of bioactive glasses have been found to stimulate the genes in osteoblasts. This effect has been found to be dose dependent. The addition of silver ions to bioactive glasses has also been investigated to produce glasses with bactericidal properties. This paper discusses how changes in the hierarchical pore structure affect the dissolution of the glass and therefore its bioactivity and rate of ion delivery and demonstrates that silver containing bioactive glass foam scaffolds can be synthesised. It was found that the rate of release of Si and Ca ions was more rapid for pore structures with a larger modal pore diameter, although the effect of tailoring the textural porosity on the rate of ion release was more pronounced. Bioactive glass scaffolds, containing 2 mol% silver, released silver ions at a rate that was similar to that which has previously been found to be bactericidal but not high enough to be cytotoxic to bone cells. PMID- 17122910 TI - Gene activation by bioactive glasses. AB - Bioactive glasses have been shown to regulate gene expression in both hard and soft tissue repair. New resorbable bioactive glass constructs are now being developed that can influence gene expression in the local environment by manipulating material properties such as the surface chemistry, topography and the release of dissolution ions. The success of these scaffolds, however, may depend upon a greater understanding of the bioactive glass stimulated gene expression pathways. This will allow the construction of tissue specific scaffolds with tailored surface chemistry, topography and ion release rates. This paper summarises the advances made in understanding gene expression in response to bioactive glasses and discusses the future steps required for further insights into these molecular mechanisms. PMID- 17122911 TI - The use of advanced diffraction methods in the study of the structure of a bioactive calcia: silica sol-gel glass. AB - Sol-gel derived calcium silicate glasses may be useful for the regeneration of damaged bone. The mechanism of bioactivity is as yet only partially understood but has been strongly linked to calcium dissolution from the glass matrix. In addition to the usual laboratory-based characterisation methods, we have used neutron diffraction with isotopic substitution to gain new insights into the nature of the atomic-scale calcium environment in bioactive sol-gel glasses, and have also used high energy X-ray total diffraction to probe the nature of the processes initiated when bioactive glass is immersed in vitro in simulated body fluid. The data obtained point to a complex calcium environment in which calcium is loosely bound within the glass network and may therefore be regarded as facile. Complex multi-stage dissolution and mineral growth phases were observed as a function of reaction time between 1 min and 30 days, leading eventually, via octacalcium phosphate, to the formation of a disordered hydroxyapatite (HA) layer on the glass surface. This methodology provides insight into the structure of key sites in these materials and key stages involved in their reactions, and thereby more generally into the behaviour of bone-regenerative materials that may facilitate improvements in tissue engineering applications. PMID- 17122912 TI - From the bioactive glasses to the star gels. AB - The improvement of bioactive glasses is one of the most important subjects in the field of hard tissue replacement. More than 30 years after being discovered by Prof. Hench, bioactive glasses still attract the attention of many researchers all over the World. With this article we want to pay homage to Prof. Hench by means of reviewing the main contribution of our research team to the field initiated by him in 1969. Our efforts, aimed to go further in the understanding of sol-gel glasses bioactivity as well as to improve the mechanical properties of bioactive materials, are explained. PMID- 17122913 TI - Progress in Raman spectroscopy in the fields of tissue engineering, diagnostics and toxicological testing. AB - This review summarises progress in Raman spectroscopy and its application in diagnostics, toxicological testing and tissue engineering. Applications of Raman spectroscopy in cell biology are in the early stages of development, however, recent publications have demonstrated its utilisation as a diagnostic and development tool with the key advantage that investigations of living cells can be performed non-invasively.Some of the research highlighted here demonstrates the ability of Raman spectroscopy to accurately characterise cancer cells and distinguish between similar cell types. Many groups have used Raman spectroscopy to study tissues, but recently increased effort has gone into single cell analysis of cell lines; the advantages being that cell lines offer ease of handling and increased reproducibility over tissue studies and primary cells. The main goals of bio-Raman spectroscopy at this stage are twofold. Firstly, the aim is to further develop the diagnostic ability of Raman spectroscopy so it can be implemented in a clinical environment, producing accurate and rapid diagnoses. Secondly, the aim is to optimise the technique as a research tool for the non invasive real time investigation of cell/material interactions in the fields of tissue engineering and toxicology testing. PMID- 17122914 TI - The future of biomedical materials. AB - The purpose of this communication is to present the author's perspectives on the future of biomedical materials that were presented at the Larry L. Hench Retirement Symposium held at Imperial College, London, in late September 2005. The author has taken a broad view of the future of biomedical materials and has presented key ideas, concepts, and perspectives necessary for the future research and development of biomedical polymers and their future role as an enabling technology for the continuing progress of tissue engineering, regenerative medicine, prostheses, and medical devices. This communication, based on the oral presentation, is meant to be provocative and generate discussion. In addition, it is targeted for students and young scientists who will play an ever-increasing role in the future of biomedical materials. PMID- 17122916 TI - Clinical applications of glass-ceramics in dentistry. AB - Glass-ceramics featuring special properties can be used as a basis to develop biomaterials. It is generally differentiated between highly durable biomaterials for restorative dental applications and bioactive glass-ceramics for medical use, for example, bone replacements. In detail, this paper presents one biomaterial from each of these two groups of materials. In respect to the restorative dental biomaterials, the authors give an overview of the most important glass-ceramics for clinical applications. Leucite, leucite-apatite, lithium disilicate and apatite containing glass-ceramics represent biomaterials for these applications. In detail, the authors report on nucleation and crystallization mechanisms and properties of leucite-apatite glass-ceramics. The mechanism of apatite nucleation is characterized by a heterogeneous process. Primary crystal phases of alpha - and beta -NaCaPO4 were determined. Rhenanite glass-ceramics represent biomaterials with high surface reactivity in simulated body fluid, SBF, and exhibit reactive behaviour in tests with bone cells. Cell adhesion phenomena and cell growth were observed. Suitable colonization and proliferation and differentiation of cells as a preliminary stage in the development of a material for bone regeneration applications was established. The authors conclude that the processes of heterogeneous nucleation and crystallization are important for controlling the required reactions in both biomaterial groups. PMID- 17122915 TI - Historic and current strategies in bone tissue engineering: do we have a hope in Hench? AB - Professors Larry Hench and Julia Polak formed the Tissue Engineering and Regenerative Medicine Centre (TERM) at Imperial College London to foster collaborations between biologists and materials scientists. Early work at the center elucidated the biomolecular interactions between primary human osteoblasts and 45S5 Bioglass . As research efforts expanded, the team discovered that the dissolution products of both 45S5 Bioglass and 58S sol-gel bioactive glasses had osteoblastic stimulatory properties. To address the shortage of appropriate cells for bone tissue engineering applications, TERM scientists also demonstrated the differentiation of embryonic stem (ES) cells to osteoblasts when treated with the dissolution products of bioactive glasses. They also found that the soluble factors ascorbic acid, beta -glycerophosphate, and dexamethasone preferentially differentiated ES cells to osteoblasts, and their combination with the dissolution products of bioactive glasses stimulated differentiation even further. Taken together, these results demonstrate the suitability of bioactive glasses as scaffolds for bone tissue engineering as they not only provide an osteoconductive and osteoproductive substrate, but also actively stimulate cells to express appropriate osteoblastic phenotypes. Professor Hench's vision to pioneer regenerative medicine research continues with the aim of developing novel therapeutics to treat musculoskeletal disability. PMID- 17122917 TI - Skeletal tissues as nanomaterials. AB - Collagen is the most abundant protein in the body and, though the fibre-forming collagens have a 'common' structure, it is adapted to perform a large range of functions-from the differing mechanical needs of tendon versus bone to forming a transparent support structure in the cornea. This perfidy also suggests that collagen could form a generic basis for a range of scaffold needs for tissue engineering or medical device coating applications. We at the London Centre for Nanotechnology--a joint venture between University College London and Imperial College--are taking a bottom-up approach having decided that many of the 'accepted dogmas' of collagen biology may not be quite as soundly based as currently held. We are using several of the tools of 'hard' nanotechnology--such as atomic force microscopy--to re-examine collagen structure with the longer term aim of using such information to design materials with appropriate physical attributes. Examples of our current research on mineralised and soft tissue collagens are presented. PMID- 17122918 TI - Zonal release of proteins within tissue engineering scaffolds. AB - The manufacture of a scaffold for tissue engineering applications that can control the location and timing of growth factor release is described. The scaffold is formed by the sintering of poly(DL-lactic acid) (P(DL)LA) microparticles, plasticized with poly(ethylene glycol) (PEG), although the method can be used for many other polymer types. The microparticles were loaded with model proteins, trypsin and horseradish peroxidase (HRP), or recombinant human bone morphogenetic protein-2 (rhBMP-2). Entrapment efficiencies above 75% were achieved using a solid-in-oil-in-water system. Controlled release of active protein was achieved for at least 30 days. Microparticles were built into protein loaded or protein-free layers and release of the protein was restricted to zones within the scaffold. Cell response to rhBMP-2 was tuneable by changing the dose of the rhBMP-2 released by varying the ratio of protein-loaded and protein-free microparticles within scaffolds. Zonal activity of rhBMP-2 on C2C12 cells was demonstrated. The scaffolds may find utility in applications where gradients of growth factors within 3D templates are required or where zonation of tissue growth is required. PMID- 17122919 TI - Surface behaviour of biomaterials: the theta surface for biocompatibility. AB - "Biomaterials" are non-living substances selected to have predictable interactions with contacting biological phases, in applications ranging from medical/dental implants to food processing to control of biofouling in the sea. More than 30 years of empirical observations of the surface behaviours of various materials in biological settings, when correlated with the contact-angle determined Critical Surface Tensions (CST) for these same materials, support the definition of the "theta surface". The "theta surface" is that characteristic expression of outermost atomic features least retentive of depositing proteins, and identified by the bioengineering criterion of having measured CST between 20 and 30 mN/m. Biomaterials applications requiring strong bioadhesion must avoid this range, while those requiring easy release of accumulating biomass should have "theta surface" qualities. Selection of blood-compatible materials is a main example. It is forecast that future biomaterials will be safely and effectively translated directly to clinical use, without requiring animal testing, based on laboratory data for CST, protein denaturation, and cell spreading alone. PMID- 17122920 TI - Synthesis and sintering of biomimetic hydroxyapatite nanoparticles for biomedical applications. AB - Synthesis of monodisperse nanoparticles with uniform morphology and narrow size distribution as achieved by nature is a challenge to materials scientists. Mimicking the process of biomineralization has led to the development of biomolecules mediated synthesis of nanoparticles that overcomes many of the problems associated with nanoparticle synthesis. Termed as biomimetics this paradigm shift in the philosophy of synthesis of materials is very advantageous for the design-based synthesis of nanoparticles. The effect of concentration of a protein named bovine serum albumin on particle size, morphology and degree of crystallinity of biomimetically synthesized hydroxyapatite particles, has been studied. Results establish 0.5% protein as the required concentration to produce 30-40 nm sized hydroxyapatite particles with an optimum degree of crystallinity as required for biomedical applications. These particles synthesized under certain stringent conditions are found to have stoichiometric calcium:phosphorus ratio of 1.67 and exhibit restricted grain growth during sintering. PMID- 17122921 TI - Macroporous bioactive glass-ceramic scaffolds for tissue engineering. AB - Highly bioactive scaffolds for tissue engineering were synthesized using a glass belonging to the SiO2-CaO-K2O (SCK) system. The glass SCK was prepared by a traditional melting-quenching route and its bioactivity was assessed by in vitro tests in a simulated body fluid (SBF). The glass was ground and sieved to obtain powders of specific size that were subsequently mixed with polyethylene particles of two different dimensions. The powders were then uniaxially pressed to obtain a crack free green compact that was thermally treated to remove the organic component and to sinter the inorganic phase. The obtained biomaterial was characterised by means of X-ray Diffraction, SEM equipped with EDS, mercury intrusion porosimetry, density measurements, image analysis, mechanical tests and in vitro evaluations. A glass-ceramic macroporous scaffold with a homogenously distributed and highly interconnected porosity was obtained. The amount and size of the introduced porosity could be tailored using various amounts of polyethylene powders of different size. PMID- 17122922 TI - Crystallinity in apatites: how can a truly disordered fraction be distinguished from nanosize crystalline domains? AB - In the last decade synthetic apatites mimicking the human natural one have been widely prepared and characterized from the physico-chemical point of view; however a shading zone is still remaining related to the evaluation and distinction of the less crystalline part, almost amorphous, and the crystallographically well ordered, nano-sized part, inside the apatite itself. Actually natural apatite forming bone tissue can include both types of crystals whose prevalence is dependent from the specific bone evolution stage and the specialized tissue performance. The quantitative description of such a combination usually represents a puzzling problem, but the result can also clarify the definition of "crystallinity in apatite" that appears still controversial. Many different synthetic apatites, including those nucleated on organic templates, were analyzed with different techniques (X-ray diffraction, transmission electron microscopy, and so on) to clarify the true nature of the disordered part. The results, manipulated by the classical methodologies devised for substances with highly perturbed structural order, led to establish that only specifically prepared amorphous calcium phosphate is really a glass, while the distorted portion coexisting with more or less crystalline regions is simply nanocrystalline. Moreover, at the conceptual limit of crystallinity tending to zero, the two models surprisingly cease to be conflicting. PMID- 17122923 TI - The effect of pressure during sintering on the strength and the fracture toughness of hydroxyapatite ceramics. AB - Hydroxyapatite (HA) is known to be biocompatible and osteoconductive, and can be synthesized chemically. The objective of the present study is to clarify the effect of pressure during sintering on the mechanical properties of HA. HA was sintered using a hot press system at a uniaxial pressure ranging from 7.81 to 62.5 MPa at a maximum temperature of 1200 degrees C with a heating rate of 10 degrees C/min. The density of the HA increased with increasing pressure and peaked at the sintering pressure of 31.2 MPa. Four-points bending tests and fracture toughness measurements with indentation method were conducted to clarify the effect of sintering pressure. Bending strength decreased at the pressure > 31.2 MPa. This result indicates that residual stress generated during sintering process became larger with increasing pressure. Fracture toughness were also lower with high density HA. PMID- 17122924 TI - Cytotoxicity study of plasma-sprayed hydroxyapatite coating on high nitrogen austenitic stainless steels. AB - Stainless steel has been frequently used for temporary implants but its use as permanent implants is restricted due to its low pitting corrosion resistance. Nitrogen additions to these steels improve both mechanical properties and corrosion resistance, particularly the pitting and crevice corrosion resistance. Many reports concerning allergic reactions caused by nickel led to the development of nickel free stainless steel; it has excellent mechanical properties and very high corrosion resistance. On the other hand, stainless steels are biologically tolerated and no chemical bonds are formed between the steel and the bone tissue. Hydroxyapatite coatings deposited on stainless steels improve osseointegration, due their capacity to form chemical bonds (bioactive fixation) with the bone tissue. In this work hydroxyapatite coatings were plasma sprayed on three austenitic stainless steels: ASTM-F138, ASTM-F1586 and the nickel-free Bohler-P558. The coatings were analyzed by SEM and XDR. The cytotoxicity of the coatings/steels was studied using the neutral red uptake method by quantitative evaluation of cell viability. The three uncoated stainless steels and the hydroxyapatite coated Bohler-P558 did not have any toxic effect on the cell culture. The hydroxyapatite coated ASTM-F138 and ASTM-F1586 stainless steels presented cytotoxicity indexes (IC50%) lower than 50% and high nickel contents in the extracts. PMID- 17122925 TI - Surface morphology and adsorbed proteins affect phagocyte responses to nano porous alumina. AB - This study evaluates human neutrophil responses to aluminum oxide membranes with different pore sizes (20 nm and 200 nm in diameter) uncoated and pre-coated with serum, collagen I, or fibrinogen. The effect of released neutrophil granule components on the survival of osteoblastic cells (MG63) bound to the alumina membranes has also been evaluated. Without protein coatings the 20 nm pore-size membranes prompt higher reactive oxygen species (ROS) production as assessed by luminol-amplified chemiluminescence than the 200 nm pore-size membranes. Such pore-size depending responses were also found on membranes pre-coated with fibrinogen, but not with collagen or serum were in fact a much lower ROS production was observed. In addition, uncoated and fibrinogen-coated membranes prompt stronger release of the granule enzymes, myeloperoxidase and elastase, than collagen or serum-coated alumina. Equally important, we found that surface mediated phagocyte activation and the subsequent release of granule components had a significant affect on the adhesion, viability and proliferation of osteoblasts. This stresses the importance of studying not only cell/surface interactions but also cell/cell interactions in wound healing and tissue regeneration processes. PMID- 17122926 TI - The properties of biomimetically processed calcium phosphate on bioactive ceramics and their response on bone cells. AB - This study looks for grounds to alter the chemical composition (phosphate, calcium, silica and carbonate), dissolution properties, structure and nanotopography of the biomimetically processed surfaces on bioactive ceramics to optimize their shown ability to influence bone cell behaviour and production of new bone. In the bone environment desirable characteristic of these materials is their ability to be remodeled by natural osteoclastic resorption. Different silica and carbonate containing calcium phosphate layers were prepared on bioactive glasses 9 (S53P4) and 1-98 (S53P2) and sol-gel processed pure silica SiO2 in C- and R-SBF (conventional and revised simulated body fluid) for varying periods of time. It was shown that in R-SBF the CaP layer formed faster compared to C-SBF. The CaP layer in the R-SBF contained more carbonate (CO3(2-)) compared to that formed with the same immersion time in C-SBF. The CaP so formed in R-SBF with faster precipitation is more amorphous than the bonelike HCA formed in C SBF. The results indicate that the most suitable surface for both osteoblasts and osteoclasts was found to be an amorphous CaP having mesoporous nanotopography and proper dissolution rate of calcium and silica. PMID- 17122927 TI - Characterization of a novel calcium phosphate composite bone cement: flow, setting, and aging properties. AB - The flow, setting, and aging characteristics of a newly developed calcium phosphate/calcium aluminate composite orthopaedic cement were studied. The effect of vibration on the flow of the cement paste was studied and found to greatly enhance placement. The setting times of this cement were dependent on temperature and decreased with increasing temperatures. At 37 degrees C, the working and setting times were 6.3 +/- 0.3 and 12.8 +/- 0.4 minutes, respectively. Hydration and conversion of the cement phases continued while specimens were stored under simulated, physiological conditions. A cumulative increase in mass of 8.23 +/- 0.65% was observed over a 14 month test period. During this time, the cement was found to expand slightly, 0.71 +/- 0.39%. X-ray diffraction was used to characterize the crystalline phases present during hydration and conversion. The calcium aluminate in the cement hydrated and formed calcium aluminate chloride hydrates, while no changes were observed in the beta-tricalcium phosphate during the testing period. PMID- 17122928 TI - Study on machinable glass-ceramic containing fluorophlogopite for dental CAD/CAM system. AB - The glass-ceramic mainly containing fluorophlogopite is one of widely used dental ceramics. In the K2O-CaO-MgO-Al2O3-SiO2-F system, a new-type glass-ceramic containing fluorophlogopite Ca-mica has been synthesized. Its crystalline was studied by XRD and EDS. The fluorophlogopite whose formula postulated K(1 - X )Ca( X/2)Mg(3)AlSi(3)O(10)F(2) was its main crystalline. The microstructure of the glass-ceramic displayed typical machinable microstructure with lath like crystals isolated and interlocking with different aspect ratio. The material also showed better bending strength (228.11 +/- 7.55 MPa). It took less than 12 minutes to fabricate a whole crown by dental CAD/CAM system with the glass ceramic. PMID- 17122929 TI - Bone-like apatite coating on Mg-PSZ/Al2O3 composites using bioactive systems. AB - A biomimetic method was used to promote bioactivity on zirconia/alumina composites. The composites were composed of 80 vol% Mg-PSZ and 20 vol% Al2O3. Samples of these bioinert materials were immersed in simulated body fluid (SBF) for 7 days on either a bed of wollastonite ceramics or bioactive glass. After those 7 days, the samples were immersed in a more concentrated solution (1.4 SBF) for 14 days. Experiments were also performed without using a bioactive system during the first stage of immersion. A bone-like apatite layer was formed on the surface of all the materials tested, using wollastonite the bioactive layer was thicker and its morphology was close to that observed on the existing bioactive systems. A thinner apatite layer consisting of small agglomerates was obtained using bioactive glass. The thickness of the ceramic layers was within the range of 15 to 30 microm. PMID- 17122930 TI - Injectable bone substitute to preserve alveolar ridge resorption after tooth extraction: a study in dog. AB - The aim of the present study was to assess the efficacy of a ready-to-use injectable bone substitute on the prevention of alveolar ridge resorption after tooth extraction. Maxillary and mandibular premolars were extracted from 3 Beagle dogs with preservation of alveolar bone. Thereafter, distal sockets were filled with an injectable bone substitute (IBS), obtained by combining a polymer solution and granules of a biphasic calcium phosphate (BCP) ceramic. As a control, the mesial sockets were left unfilled. After a 3 months healing period, specimens were removed and prepared for histomorphometric evaluation with image analysis. Histomorphometric study allowed to measure the mean and the maximal heights of alveolar crest modifications. Results always showed an alveolar bone resorption in unfilled sockets. Resorption in filled maxillary sites was significantly lower than in control sites. Interestingly, an alveolar ridge augmentation was measured in mandibular filled sockets including 30% of newly formed bone. It was concluded that an injectable bone substitute composed of a polymeric carrier and calcium phosphate can significantly increase alveolar ridge preservation after tooth extraction. PMID- 17122931 TI - In vitro testing of Nd:YAG laser processed calcium phosphate coatings. AB - Nd:YAG laser cladding is a new method for deposition of a calcium phosphate onto metallic surfaces of interest in implantology. The aim of this study was to compare the biologic response of MG-63 human osteoblast-like cells grown on Ti 6Al-4V substrates coated with a calcium phosphate layer applied using different methods: plasma spraying as reference material and Nd:YAG laser cladding as test material. Tissue culture polystyrene was used as negative control. The Nd:YAG laser clad material showed a behaviour similar to the reference material, plasma spray, respective to cell morphology (SEM observations), cell proliferation (AlamarBlue assay) and cytotoxicity of extracts (MTT assay). Proliferation, as measured by the AlamarBlue assay, showed little difference in the metabolic activity of the cells on the materials over an 18 day culture period. There were no significant differences in the cellular growth response on the test material when compared to the ones exhibited by the reference material. In the solvent extraction test all the extracts had some detrimental effect on cellular activity at 100% concentration, although cells incubated in the test material extract showed a proliferation rate similar to that of the reference material. To better understand the scope of these results it should be taken into account that the Nd:YAG clad coating has recently been developed. The fact that its in vitro performance is comparable to that produced by plasma spray, a material commercially available for more than ten years, indicates that this new laser based method could be of commercial interest in the near future. PMID- 17122932 TI - Bond-coating in plasma-sprayed calcium-phosphate coatings. AB - The influence of bond-coating on the mechanical properties of plasma-spray coatings of hydroxyatite on Ti was investigated. Plasma-spray powder was produced from human teeth enamel and dentine. Before processing the main apatite coating, a very thin layer of Al2O3/TiO2 was applied on super clean and roughened, by Al2O3 blasting, Ti surface as bond-coating. The experimental results showed that bond-coating caused significant increase of the mechanical properties of the coating layer: In the case of the enamel powder from 6.66 MPa of the simple coating to 9.71 MPa for the bond-coating and in the case of the dentine powder from 6.27 MPa to 7.84 MPa, respectively. Both tooth derived powders feature high thermal stability likely due to their relatively high content of fluorine. Therefore, F-rich apatites, such those investigated in this study, emerge themselves as superior candidate materials for calcium phosphate coatings of producing medical devices. The methods of apatite powder production and shaping optimization of powder particles are both key factors of a successful coating. The methods used in this study can be adopted as handy, inexpensive and reliable ways to produce high quality of powders for plasma spray purposes. PMID- 17122933 TI - Precipitation of octacalcium phosphates on artificial enamel in artificial saliva. AB - The natural saliva samples were collected from 30 Japanese men and women aged between 20 and 30, and the inorganic components in the saliva were analyzed chemically. Artificial saliva (AS) was prepared based on the chemical analysis using chemical reagents. The calcium/phosphate molar ratio of the AS was 0.28 with pH=7.0. Artificial enamel (AE) was prepared by sintering hydroxyapatite powder at 1,200 degrees C. The AE was placed in the AS at 37 degrees C. XRD, SEM and EDX investigated precipitation on the AE. The precipitation was always identified as being OCP. As decrease in the pH of the surface of the AE and increase in the concentration of phosphate ions were observed by chemical analysis, it was concluded that the OCP occurs by the pH decrease and phosphate ion concentration increase on the surface of AE. PMID- 17122934 TI - Calcium phosphate fibres synthesized from a simulated body fluid. AB - The biomimetic coating method was used for fabricating calcium phosphate fibres for biomedical applications such as bone defect fillers. Natural cotton substrate was pre-treated with phosphorylation and a Ca(OH)2 saturated solution. The pre treated samples were then soaked in simulated body fluid (SBF) of two different concentrations, 1.5 times and 5.0 times the ion concentration of blood plasma. The cotton was then burnt out via sintering of the ceramic coating at 950 degrees C, 1050 degrees C, 1150 degrees C, and 1250 degrees C. The results demonstrated that osteoblastic cells were able to cover the entire surface cotton fibres, and the cell coverage appeared to be independent of surface roughness and Ca/P ratio of fibres. PMID- 17122935 TI - Liver transplantation in transthyretin amyloidoses. When the neurological benefit outweighs the risk. PMID- 17122936 TI - Visual evoked potentials and diabetic polyneuropathy. PMID- 17122937 TI - Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL): a proteiform neurological disease of expanding importance. Reasons for establishing an Italian Registry. PMID- 17122938 TI - High plasma creatine kinase: review of the literature and proposal for a diagnostic algorithm. AB - The condition of persistently high plasma CK levels is frequently encountered in asymptomatic patients with normal neurological examination. This condition may be the unique manifestation of several neuromuscular disorders, whose diagnosis is now possible using new diagnostic techniques. However, even if these patients are intensely investigated, specific diagnoses are not always forthcoming. Because of the lack of a widely accepted diagnostic protocol, hyperCKaemia in asymptomatic subjects is a potentially difficult clinical problem. In this paper we review the literature on conditions associated with variations in plasma CK levels and the literature on investigations carried out in asymptomatic persons with high CK to identify neuromuscular diseases. In the light of these data, and the deliberations of a working group of the Italian Association of Myology, we propose a diagnostic algorithm to guide the diagnostic work-up of persons presenting with persistently high levels of plasma CK. This algorithm has been discussed and approved by the Committee of the Italian Association of Myology. PMID- 17122939 TI - Effect of nitric oxide on lymphocytes from sporadic amyotrophic lateral sclerosis patients: toxic or protective role? AB - Markers of oxidative and nitrosative stress have been found in spinal cord, cortex, cerebrospinal fluid and plasma of patients affected by amyotrophic lateral sclerosis (ALS), a fatal disorder characterised by progressive motor neuron degeneration. In this study, we investigated the effect of the NO releasing agent, diethylamine NONOate (NONO), on lymphocytes from patients affected by the sporadic form of ALS (SALS) and controls by flow cytometry. In the same experimental conditions we investigated the expression of the antioxidant proteins, Bcl-2 and SOD1. Incubation with NONO induced cell damage in control lymphocytes but did not further damage the already affected untreated SALS lymphocytes. The incubation with NONO induced a time-dependent decrease of Bcl-2 and SOD1 in control lymphocytes. Surprisingly, in SALS lymphocytes the NONO treatment increased the expression of these proteins, which in basal conditions was depressed compared to control lymphocytes. PMID- 17122940 TI - Iatrogenic intracranial pseudoaneurysms: neuroradiological and therapeutical considerations, including endovascular options. AB - Intracranial pseudoaneurysms represent a potentially fatal complication of intracranial surgery. Our purpose is to describe their neuroradiological characteristics, prognostic features and possible treatment. Eight cases of postsurgical intracranial pseudoaneurysms have been observed at our institution since 1988. Four were observed following transsphenoidal (TS) surgery and four after pterional craniotomies. Two types of iatrogenic pseudoaneurysms were observed: "fusiform", probably due to weakening of the adventitia during surgical peeling of the tumour from the artery (three cases) and "saccular", occurring after a more focal or complete laceration of the vessel (five cases), more often after TS surgery. A thorough preoperative neuroradiological examination may identify anatomical conditions at risk for development of this severe complication. Postoperative neuroradiological follow-up is mandatory in cases in which unusual bleeding has occurred during the perioperative period, but absence of bleeding does not exclude the possible development of a pseudoaneurysm. Endovascular treatment of pseudoaneurysms represents a safe and durable procedure, specifically in those cases in which damage to the carotid siphon occurred during TS surgery. PMID- 17122941 TI - Luminance and chromatic visual evoked potentials in type I and type II diabetes: relationships with peripheral neuropathy. AB - The objective of the study was to investigate the subclinical visual deficit in type I and II diabetes, and its relationship with peripheral neuropathy. Thirty two healthy volunteers, 20 patients with type I diabetes and 30 patients with type II diabetes were studied in a clinical neurophysiology setting. Luminance (VEPs) and chromatic visual evoked potentials (CVEPs) were recorded, with white black, grey-black, red-green and blue-yellow sinusoidal gratings. The peak latencies of the VEP positive wave and CVEP negative wave were recorded. Ten patients with type I and 8 with type II diabetes had peripheral neuropathy. VEPs were slower in patients with type II diabetes and CVEPs were slower in patients with type I and type II diabetes than in controls. Blue-yellow CVEPs were slower in type II than in type I diabetes. VEPs and red-green CVEPs were slower in patients with diabetes with neuropathy than in those without. In conclusion, we found that visual system impairment differs in diabetes with and without peripheral neuropathy. PMID- 17122942 TI - Silent brain embolism during transcatheter closure of patent foramen ovale: a transcranial Doppler study. AB - The aim of this report is to quantify the amount of spontaneous microembolism detected in brain vessels by transcranial Doppler (TCD) during transcatheter closure of right-to-left shunt (RLS). We examined 29 patients who had had a stroke or a transient ischaemic attack (17 females and 12 males; mean age 45+/-15 years). They all underwent TCD monitoring during the procedure and microembolic signals (MES) were recorded. Detection of MES was distributed as follows: during femoral catheterisation in 8 patients (25%), during atrial catheterisation in 5 patients (17%), during transeptal crossing in 14 patients (48%), during left disc opening in 28 patients (96%) and during right disc opening in 7 patients (24%). The highest rates of MES were observed during left disc opening and less during transeptal crossing with an average count of 31 (range 3-135) and 3 (range 1-18) respectively. Brain embolism occurs throughout the procedure after femoral catheterisation for PFO closure. Our results indicate that the majority of MES reached the brain during the opening of the left disc in the left atrium: 28/29 patients exhibited MES with an average of 31 (3-135), thus supporting the notion that gas embolism accounted for the findings. PMID- 17122943 TI - Stroke Unit care in Italy. Results from PROSIT (Project on Stroke Services in Italy). A nationwide study. AB - The future challenge for improving stroke patients' outcome will be to implement new Stroke Units (SUs) worldwide. However the best SU model remains uncertain. The aim of this study was to evaluate the number of SUs and the quality characteristics of acute stroke care in Italy. We conducted a SU survey in Italy, interviewing the directors of the hospital wards that discharged at least 50 acute stroke patients a year. A SU was defined as an acute ward area with stroke dedicated beds and staff. To compare the quality of care provided in SUs with that in general wards (GWs) we investigated the characteristics of five domains: hospital setting, unit setting, staffing, process of care and diagnostic investigations. We identified 68 SUs and 677 GWs. Multivariate logistic regression analyses demonstrated that SUs compared to GWs had higher quality scores in unit setting (ROC area=0.9721), staffing (ROC area=0.8760) and care organisation (ROC area=0.7984). The hospital setting (ROC area=0.7033) and the availability of rapid diagnostic investigations (ROC area=0.7164) had lower power in discriminating SU from GW. In Italy in 2003/04 only 9% of the hospital services had organised SU care. The study demonstrated that SUs admitted more than 100 patients per year, had more monitoring equipment and staffing time, and practised multidisciplinary meetings and early mobilisation. The utility of these structural and performance characteristics needs validation from outcome studies. PMID- 17122944 TI - The role of formative courses in the professional integration of patients with epilepsy. An Italian experience. AB - The aim of the present paper was to describe some formative projects on work integration of patients with epilepsy and mild cognitive deficits, so that similar initiatives may be promoted. The different phases of the formative projects were described: the inclusion criteria, the selection of participants, the assessment instruments, the courses themselves and the final results in terms of working experience. Thirteen participants were selected for each course. Patients were treated in different Lombardy Epilepsy departments, which addressed them to the Epilepsy Center, S. Paolo Hospital, Milan, where the selection took place. Seventy percent of the participants in the formative courses found a job in 1 year, even though most of the positions were temporary. The work integration seemed to be mainly related to the level of intelligence and to verbal memory. We stress the importance of organising similar courses in the future, in the context of wider networks. This will help more patients in acquiring both economic and psychological autonomy from their families. PMID- 17122945 TI - Tumour-associated epilepsy: clinical impact and the role of referring centres in a cohort of glioblastoma patients. A multicentre study from the Lombardia Neurooncology Group. AB - Epilepsy in high-grade glioma patients is a major concern, mainly as regards indications to treatment and best choice; toxicities, and pharmocokinetic and pharmacodynamic interactions of drugs. All these generally unsolved problems complicate patients' quality of life and interfere with the evaluation of response criteria in clinical trials. A prospective, multicentre data collection on 132 adult newly diagnosed, histologically proven glioblastomas from 9 Lombardy hospitals collected in the same database during a one-year period was recently published. From this database we report epidemiological and clinical characteristics in epilepsy-symptomatic (31%) glioblastoma patients vs. the group with other presenting symptoms (69%). We analyse demographic and clinico radiological features, timing of onset and the course of seizures, and modalities of treatment in the two groups of patients. No statistically significant differences were observed between the two groups as regards age, site of lesion(s), extent of surgery and survival in relation to anticonvulsant treatment status or pharmacokinetic properties of drugs. PMID- 17122946 TI - Fatal cerebral haemorrhage after liver transplantation in a patient with transthyretin variant (gly53glu) amyloidosis. AB - Transthyretin (TTR) amyloidosis is a form of systemic amyloidosis caused by an amyloidogenetic TTR variant. The most common mutant forms of TTR are mainly produced by the liver and therefore orthotopic liver transplantation (OLT) is currently accepted as the only known curative treatment. We describe the clinical and pathological features of a patient with TTR variant (gly53glu) with TTR amyloid infiltration of the leptomeningeal vessels in whom fatal cerebral haemorrhage occurred two months after OLT, soon after severe viral pneumonia. PMID- 17122947 TI - Asymmetric Guillain-Barre syndrome. AB - Guillain-Barre syndrome (GBS) is a heterogeneous disorder according to clinical, electrophysiological, immunologic and pathologic findings. It has usually been considered as an immune-mediated polyneuropathy clinically characterised by acute symmetric muscle weakness and areflexia. We describe a patient who, after a Campylobacter jejuni infection, developed an acute motor-sensory neuropathy with marked and persistent asymmetry of clinical and electrophysiological findings. He had a high titre of anti-GM1 IgG antibodies and cytoalbuminologic dissociation and was responsive to intravenous immunoglobulins. Investigations and three years of follow-up excluded mimics of GBS. Tendon areflexia has recently been challenged as a mandatory diagnostic criterion in GBS; likewise marked and persistent motor asymmetry does not exclude the diagnosis of GBS. PMID- 17122948 TI - Leukoencephalopathy as a rare complication of hepatitis C infection. AB - We report the case of a 64-year-old female patient with hepatitis C infection (HCV), who developed Sjogren's disease and sensory peripheral neuropathy. Clinical conditions worsened over three years with central nervous system involvement characterised by transient third cranial nerve paresis and mild selective impairment of attention and memory. Brain magnetic resonance imaging showed diffuse periventricular and lobar white matter hyperintensity. Laboratory findings included mixed cryoglobulinaemia (type II), cryocrit 1.47%, low serum levels of complement C4 and high levels of rheumatoid factor, HCV 1b genotype, high HCV mRNA levels in serum and cerebrospinal fluid. Skin biopsy showed evidence of vasculitis. After one year of plasmapheresis, immunosuppressant therapy and occasional corticosteroid treatment, neurological symptoms improved, skin biopsy changed and inflammation parameters normalised, suggesting that neurological symptoms might be related to the high levels of mixed cryoglobulins. PMID- 17122949 TI - Vertebral artery dissection in a child. Is "spontaneous" still an appropriate definition? AB - Though a rare cause of stroke in the general population, in almost one quarter of young patients affected by stroke cervical artery dissection (CAD) is the underlying cause. Among these cases "spontaneous" dissections, intended as non traumatic, represent about 34% of posterior circulation arterial dissection in patients aged less than 18 years. We here describe the case of a seven-year-old boy who developed a spontaneous vertebral artery dissection (VAD) leading to occipito-mesial, thalamo-capsular and cerebellar infarction. Once a traumatic origin was excluded, clinical history and laboratory findings were further analysed: fever associated with tonsillitis during the previous week, raised inflammatory indices, a throat culture positive for beta haemolysing Streptococcus group A and high titres of streptococcal antibodies were found. This case suggests that patients with CAD referred as spontaneous deserve extensive analysis. Subjects presenting with a dissection and an underlying infection are likely to have a hyperinflammatory response (although further experience is needed). In these patients immediate start of antibiotic therapy, treatment with anti-inflammatory drugs and further a lifelong prophylaxis with antibiotics before any invasive procedure are strongly recommended. PMID- 17122951 TI - NMR assignment of the human spliceosomal 15.5K protein. PMID- 17122952 TI - Resonance assignments of the alpha subunit of human eukaryotic initiation factor 2 (heIF2alpha). PMID- 17122953 TI - Advances in the management of corticosteroid-induced osteoporosis with bisphosphonates. AB - Corticosteroids are widely used as anti-inflammatory and immunosuppressive agents to treat a variety of chronic conditions. Long-term (>1 year) corticosteroid use can lead to bone loss, and therefore, osteopenia or osteoporosis. Corticosteroid induced osteoporosis (CIO) leads to increased bone fragility and subsequently fractures, which, in turn, lead to a loss of physical, emotional and social health for the patient and increased costs for healthcare providers. A wealth of data exists demonstrating the efficacy of the oral bisphosphonates, etidronate, alendronate and risedronate in increasing bone mineral density in patients with CIO or preventing bone loss in patients commencing corticosteroid therapy. Data regarding fracture prevention are less clear, as statistically significant reductions in the incidence of fractures have only been reported for patient subgroups or meta-analyses. However, many treatment guidelines recommend the use of oral bisphosphonates for the prevention and treatment of CIO. These guidelines are, however, not reflected in prescribing practice, and the majority of patients do not receive adequate concomitant therapy. This review summarizes the available data for bisphosphonates in CIO. Therapeutic adherence with oral bisphosphonates is an issue, with approximately 50% of patients discontinuing therapy within the first year. The primary reasons for this are poor gastrointestinal tolerability and the frequency with which complex dosing requirements must be followed. The inconvenience of taking daily or weekly bisphosphonate therapy is of particular importance in patients with CIO who may be regularly taking several other medications. Data obtained in studies with ibandronate indicate that bisphosphonate administration by rapid intravenous injection provides an effective, well-tolerated and practical alternative to current oral regimens in the management of patients with CIO. PMID- 17122954 TI - Facial rejuvenation and light: our personal experience. AB - The treatment of ageing skin remains a very hot topic, and many systems have been reported as having varying degrees of success. Nonablative lasers were developed to avoid the problematic and uncomfortable sequelae following laser ablative resurfacing, and while there was no downtime, there was also poor patient satisfaction. The same was true of the intense pulsed light systems. The use of different modalities in various combinations was found to offer much better results, however, such as a 595-nm pulsed dye laser followed by a 1,450-nm diode laser, and so on, all used at subablative thresholds. The recent entry of blue and infrared tunable plasma light and light-emitting diodes into the skin rejuvenation arena has attracted a great deal of attention. The authors suggest that no single modality can accomplish all the complex events required for effective skin rejuvenation, suggest that combination phototherapy is the best approach combined with an adjunctive epidermal care regimen, and demonstrate their development of this methodology. PMID- 17122955 TI - Calcium-PTH-vitamin D axis in older patients with hip fracture. PMID- 17122956 TI - Therapeutic potential of superoxide dismutase (SOD) for resolution of inflammation. AB - Neutrophils play essential roles in several inflammatory reactions. Oxidant/antioxidant imbalance is thought to be partially involved in the pathogenesis of the disorders. Under the conditions of oxidative stress, superoxide dismutase (SOD) acts as an endogenous cellular defense system to degrade superoxide (O2-) into oxygen and hydrogen peroxide. Therefore, SOD is potentially useful as a therapeutic agent for treatment of inflammatory disorders. A further mechanism that may contribute to the efficacy of SOD is the regulation of neutrophil apoptosis. For the resolution of inflammation, the activated neutrophils must be safely removed by apoptosis. Neutrophil apoptosis has been suggested as a possible target for the control of neutrophil-mediated tissue injury. Exogenously added SOD induces neutrophil apoptosis, and hydrogen peroxide has been suggested to be a possible major mediator of ROS-induced neutrophil apoptosis in a caspase-dependent manner. If the drug can be delivered efficiently to the inflammatory site, SOD may be useful as an inhibitory mediator of neutrophil-mediated inflammation. PMID- 17122957 TI - Fasudil, a Rho-kinase inhibitor, inhibits leukocyte adhesion in inflamed large blood vessels in vivo. AB - OBJECTIVE AND DESIGN: Emerging data suggest that Rho-kinase signaling may regulate numerous aspects of inflammatory reactions. Herein, we investigated the role of Rho-kinase in inflammatory interactions between leukocytes and the endothelium in femoral arteries and veins in vivo. MATERIAL AND METHODS: Mice were injected with lipopolysaccharide (LPS) and Rho-kinase was inhibited by pre treatment with fasudil, which is a highly selective inhibitor of Rho-kinase. Six hours after LPS challenge, intravital fluorescence microscopy of the femoral vessels was performed and leukocyte-endothelium interactions were visualized after in vivo staining with rhodamine 6G. RESULTS: LPS increased leukocyte rolling and adhesion in femoral arteries and veins. Pre-treatment with fasudil had no effect on leukocyte rolling but significantly decreased venular leukocyte adhesion by 85% and completely abrogated leukocyte adhesion in femoral arteries in endotoxin-treated mice. CONCLUSIONS: We conclude that Rho-kinase signaling regulates LPS-induced leukocyte adhesion in femoral arteries and veins in vivo and that inhibition of Rho-kinase may be useful in the treatment of pathological inflammation in large blood vessels of the vascular system. PMID- 17122958 TI - Manipulation of the induction of adjuvant arthritis in Sprague-Dawley rats. AB - OBJECTIVE: To investigate the roles of various variables in the induction of adjuvant-induced arthritis (AIA) in the outbred Sprague-Dawley (SD) rats, and further characterize its arthritic features by comprehensive examinations. METHODS: The roles of different preparative techniques, inoculation routes and doses of Mycobacterium tuberculosis (MT) suspension as well as the sex preference in the induction of AIA were comparatively studied using clinical assessment. The hind paws of animals were analyzed by radiological and histological examinations. The serum levels of cytokines interleukin (IL)-1beta, IL-6, and tumor necrosis factor (TNF)-alpha were determined by ELISA. RESULTS: The particle size and dose of MT played a dominant role in the induction and severity of AIA. Male rats manifested markedly more severe arthritic signs than female rats. After subcutaneously inoculated with 500 microg MT, male rats developed pronounced arthritis with 100% incidence and low variable clinical signs. Even using only 62.5 microg MT, AIA was efficiently induced in male rats and characterized by upregulated expression profiles of IL-1beta, IL-6 and TNF-alpha. CONCLUSIONS: Since outbred SD rats are much cheaper and more readily available than Lewis rats, this well-developed SD rat AIA model is an efficient and cost-effective arthritis model available for screening novel anti-arthritic agents. PMID- 17122959 TI - Anti-tumor necrosis factor alpha F(ab')2 antibody fragments protect in murine polymicrobial sepsis: concentration and early intervention are fundamental to the outcome. AB - BACKGROUND: Negative results are frequent using anti-TNFalpha antibodies in sepsis models and clinical trials. METHODS AND RESULTS: Different prophylactic doses of anti-TNFalpha F(ab')2 antibody fragments were compared for the prevention of death by sepsis induced by cecal ligation and puncture (CLP) in mice. High (10 mg/kg) and very low (0.01 and 0.1 mg/kg) concentrations of anti TNFalpha antibody fragments were not the most adequate for treating polymicrobial sepsis, since they did not significantly improve survival. To the contrary, intermediate doses (1 mg/kg) significantly protected the challenged animals. Protective activity was also observed when administration of the antibody fragments was initiated early (up to 30 min) after CLP. CONCLUSIONS: These results suggest that in processes where excessive production of cytokines is involved, the aim should be to return them to their physiologically acting range but not to inhibit their production. The timing of initiating therapy should also be considered in order to maximize the possible benefits. PMID- 17122960 TI - Protective effects of propofol on lipopolysaccharide-activated endothelial cell barrier dysfunction. AB - BACKGROUND: Propofol has been widely used in intravenous anesthesia. It possesses antioxidant and immunomodulating effects. This study aimed to investigate whether propofol may attenuate lipopolysaccharide (LPS)-induced endothelial cell barrier dysfunction and the possible mechanisms of such modulation. METHODS: Cultured human umbilical vein endothelial cells (HUVECs) were used to assess the following treatments: (i) no additives (negative control), (ii) LPS alone (1 and 10 microg/ml), (iii) propofol alone (20 microg/ml), (iv) intralipid (a solvent of propofol) alone (20 microg/ml), (v) LPS (10 microg/ml) combination with propofol (4 and 20 microg/ml) and (vi) LPS (10 microg/ml) combination with intralipid (20 microg/ml). Changes of cell permeability and filamentous actin (F-actin) were determined. Expression of nitrotyrosine proteins and activity of nuclear factor kappaB (NF-kappaB) were analyzed by Western blot and immunocytochemistry. Expression of endothelial nitric-oxide synthase (eNOS) and inducible nitric-oxide synthase (iNOS) were analyzed by reverse transcriptase-polymerase chain reaction. RESULTS: LPS markedly increased the permeability of endothelial cells, the formation of peroxynitrite and depolymerization of F-actin in HUVECs. LPS also significantly increased mRNA of iNOS, protein level of NF-kappaB and decreased mRNA of eNOS (P < 0.05). Propofol at both concentrations (4 and 20 microg/ml) significantly inhibited the LPS-induced increase in cell permeability and alteration in F-actin organization. Propofol also reduced the LPS-enhanced iNOS mRNA and NF-kappaB protein levels whilst it increased eNOS mRNA expression (P < 0.05). CONCLUSION: This study demonstrated that propofol, both at therapeutic concentrations and 5 times therapeutic concentrations, inhibited NF-kappaB activation in LPS-stimulated endothelial cells and was found to protect endothelial cells against LPS-induced barrier dysfunction. PMID- 17122961 TI - Histamine induces interleukin-6 expression in the human synovial sarcoma cell line (SW982) through the H1 receptor. AB - METHODS: The effect of histamine on inositol phosphate generation and interleukin 6 (IL-6) release from the synovial sarcoma cell line SW982 was investigated. RESULTS: SW982 cells express functional H1 and H2 receptors. The H1 receptor antagonist [3H]-mepyramine binds to membranes from SW982 cells with high affinity and the binding was potently blocked by H1 antagonists. Histamine potently stimulated phosphoinositide (PI) hydrolysis and Ca2+ mobilization with EC50 of 4.0 +/- 0.8 microM and 1.3 +/- 0.6 microM respectively and these activities were blocked by the H1 selective antagonist mepyramine. Histamine (EC50 = 1.8 +/- 1.1 microM) stimulated the release of IL-6 that was attenuated by selective H1 antagonists. The PKC inhibitor, GF1090203X, blocked the histamine stimulated IL-6 release. The H2 selective antagonist, cimetidine, had no significant effect on histamine-induced PI turnover, Ca2+ mobilization and IL-6 release. CONCLUSION: We conclude that histamine stimulates IL-6 release from SW982 cells by binding to the H1 receptor and this is coupled to the PI/PKC signal transduction pathway. Development of an H1 antagonist that inhibits the release of IL-6 from synoviocytes may be beneficial for the treatment of inflammatory joint disease. PMID- 17122962 TI - Antiinflammatory effects of natural tetranortriterpenoids isolated from Carapa guianensis Aublet on zymosan-induced arthritis in mice. AB - OBJECTIVE: We investigated the antiinflammatory properties of a derived fraction of tetranortriterpenoids (TNTP) obtained from the seeds of Carapa guianensis Aublet. MATERIAL AND METHODS: Zymosan-induced arthritis and pleurisy in Swiss and C57/Bl6 mice (n = 10 per group). Western blot analysis was performed to analyze nuclear factor-kappaB (NFkappaB) translocation in mice peritoneal macrophages stimulated in vitro with zymosan (500 microg/ml). ELISA was performed to evaluate cytokine levels in knee joints. Values of p 0.05). The posttraining times differed between the four groups. Post hoc analyses showed statistically significant differences (p < 0.05) between the participants trained with both trainers and the control subjects. CONCLUSIONS: Our data demonstrate that the combination of virtual reality training and inanimate box training leads to better laparoscopic skill acquisition than either training method alone or no training at all. Optimal preclinical laparoscopic training should incorporate both virtual reality trainers and inanimate box trainers. PMID- 17122976 TI - Open vs laparoscopic partial posterior fundoplication. A prospective randomized trial. AB - OBJECTIVE: This study compares outcomes following open and laparoscopic partial posterior fundoplication for gastroesophageal reflux disease concerning perioperative course, postoperative complications, symptomatic relief, recurrent disease, and the need for reinterventional surgery. METHODS: A prospective randomized trial was performed. Pre- and postoperative testing included endoscopy, esophageal function testing, patient questionnaire, and clinical assessment. Patients were followed for three years. MATERIALS: Ninety-three patients were randomized to open and 99 to laparoscopic surgery. RESULTS: Complication rates were higher, and length of stay (LOS) [5 (3-36) vs 3 (1-12) days] and time off work [42 (12-76) vs 28 (0-108) days] was longer in the open group (p < 0.01). Early side effects and recurrences were more common (p < 0.05) in the laparoscopic group. One patient in the open group and 8 patients in the laparoscopic group required surgery for recurrent disease and 7 patients required surgery for incisional hernias after open surgery. Overall, at one and three years, there were no differences in patient-assessed satisfactory outcome (93.5/93.5 vs 88.8/90.8%) or reflux control (p = 0.53) between the open and laparoscopic groups. CONCLUSIONS: The finding of fewer general complications, shorter length of stay and recovery, similar need for reoperations, and comparable 3-year outcomes, makes the laparoscopic approach the primary choice when considering surgical options for the treatment of gastroesophageal reflux disease (GERD). PMID- 17122977 TI - Patent processus vaginalis in the adult as a risk factor for the occurrence of indirect inguinal hernia. AB - BACKGROUND: Inguinal hernias are a common entity with nearly 31,000 repairs annually in The Netherlands and over 800,000 in the USA. The aim of the present study is to determine whether a laparoscopically diagnosed patent processus vaginalis (PPV) is a risk factor for the development of groin hernia. METHODS: The study population was originally composed of 599 consecutive cases (189 male, 32%) of laparoscopic transperitoneal surgery for different indications performed in 4 teaching hospitals in The Netherlands between November 1998 and February 2002. During laparoscopy, the deep inguinal ring was inspected bilaterally. The PPV group was compared with the obliterative processus vaginalis (OPV) group. RESULTS: After a mean follow-up of 5.5 years, the studied population consisted of 337 cases (94 male, 28%). In this study 12% of the studied population appeared to have PPV in adult life. The percentage PPV of our study group is much higher than the percentage of hernia repairs performed in the Dutch population. A greater proportion (12%) of hernia repairs in the PPV group was found as compared with the OPV group (3%). The chance of developing an inguinal hernia within 5.3 years is four times higher in the group with PPV. No significant correlation between age and the prevalence of PPV was observed. CONCLUSION: This study demonstrates that PPV is an etiologic factor and a risk factor for acquiring an indirect inguinal hernia in adults. PMID- 17122978 TI - Effect of partial and total laparoscopic fundoplication on esophageal body motility. AB - BACKGROUND: Abnormal esophageal body motility often accompanies gastroesophageal reflux disease (GERD). Although the effect of surgery on the pressure and behavior of the lower esophageal sphincter (LES) has been extensively studied, it still is unclear whether a successful fundoplication improves esophageal peristalsis. METHODS: The pre- and postoperative esophageal manometries of 71 patients who underwent a successful laparoscopic fundoplication (postoperative DeMeester score < 14.7) were reviewed. The patients were grouped according to the type of fundoplication (partial vs total) and preoperative esophageal peristalsis (normal vs abnormal): group A (partial fundoplication and abnormal esophageal peristalsis; n = 16), group B (total fundoplication and normal peristalsis; n = 41), and group C (total fundoplication and abnormal peristalsis; n = 14). RESULTS: The LES pressure was increased in all the groups. A significant increase in amplitude of peristalsis was noted in groups A and C. Normalization of peristalsis was achieved in 31% of the group A patients and 86% of the group C patients. No changes occurred in group B. CONCLUSIONS: Laparoscopic fundoplication increased LES pressure and the strength of esophageal peristalsis in patients with abnormal preoperative esophageal motility. A total fundoplication resulted in normalization of peristalsis in the majority of patients. PMID- 17122979 TI - The influence of different training schedules on the learning of psychomotor skills for endoscopic surgery. AB - BACKGROUND: Psychomotor skills for endoscopic surgery can be trained with virtual reality simulators. Distributed training is more effective than massed training, but it is unclear whether distributed training over several days is more effective than distributed training within 1 day. This study aimed to determine which of these two options is the most effective for training endoscopic psychomotor skills. METHODS: Students with no endoscopic experience were randomly assigned either to distributed training on 3 consecutive days (group A, n = 10) or distributed training within 1 day (group B, n = 10). For this study the SIMENDO virtual reality simulator for endoscopic skills was used. The training involved 12 repetitions of three different exercises (drop balls, needle manipulation, 30 degree endoscope) in differently distributed training schedules. All the participants performed a posttraining test (posttest) for the trained tasks 7 days after the training. The parameters measured were time, nontarget environment collisions, and instrument path length. RESULTS: There were no significant differences between the groups in the first training session for all the parameters. In the posttest, group A (training over several days) performed 18.7% faster than group B (training on 1 day) (p = 0.013). The collision and path length scores for group A did not differ significantly from the scores for group B. CONCLUSION: The distributed group trained over several days was faster, with the same number of errors and the same instrument path length used. Psychomotor skill training for endoscopic surgery distributed over several days is superior to training on 1 day. PMID- 17122980 TI - Laparoscopic liver resection using radiofrequency coagulation. AB - BACKGROUND: The use of radiofrequency (RF) energy has been described to perform open liver resection safely and with minimal blood loss. Yet no data are available on the potential contribution of RF energy to the limitation of intraoperative blood loss during laparoscopic liver resection (LLR). The aim of this prospective, nonrandomized study was to investigate the potential contribution of RF energy to the limitation of intraoperative blood loss in patients undergoing LLR. METHODS: Forty-five patients [male/female ratio 22/23, age 57 years (26-80)] underwent LLR. Eleven benign and 47 malignant lesions (mostly colorectal metastases) were resected. Median number [1 (1-3)] and maximum diameter [40 mm (8-170)] of tumors as well as median tumor free margins [10 mm (1 30)] were comparable in patients undergoing LLR with (20 patients) or without (25 patients) RF-assistance. Thirty-eight minor (< or = 2 segments) and 9 major (> 3 segments) resections were performed. Eighteen patients simultaneously underwent additional surgery. RESULTS: No mortality occurred. Median intraoperative blood loss was 200 (5-4000) ml and was similar in patients undergoing LLR with or without RF-assistance. The type of surgical procedure was a determinant for the amount of intraoperative blood loss (p = 0.0002). Significant bleeding occurred from large hepatic vessels at major resections. Median operation time was 115 (45 360) minutes. RF-assistance didn't seem to reduce perioperative morbidity. CONCLUSIONS: LLR can be performed with minimal intraoperative blood loss, which is determined by the type of hepatectomy. Significant intraoperative bleeding occurs from large hepatic vessels during major resections. RF-assisted parenchymal transection in LLR doesn't seem to reduce blood loss, operation time, or perioperative morbidity. PMID- 17122981 TI - Routine laparoscopic ultrasound can significantly reduce the need for selective intraoperative cholangiography during cholecystectomy. AB - BACKGROUND: The use of intraoperative cholangiography (IOC), routinely rather than selectively, during laparoscopic cholecystectomy (LC) is controversial. Recent findings have shown laparoscopic ultrasound (LUS) to be safe, quick, and effective not only for screening of the bile duct for stones, but also for evaluating the biliary anatomy. This study aimed to evaluate, on the basis of the LC outcome and the cost of LUS and IOC, whether and how much the routine use of LUS would be able to reduce the need for IOC. METHODS: During LC, LUS was used routinely to screen the bile duct for stones and to evaluate the biliary anatomy, whereas IOC was used selectively only when LUS was unsatisfactory or unsuccessful. RESULTS: For 193 (96.5%) of 200 patients, LUS was completed successfully, whereas IOC was needed for 7 patients (3.5%). Bile duct stones were identified in 20 patients (10%). For the detection of bile duct stones, LUS yielded 19 true-positive, 175 true-negative, 0 false-positive, and 1 false negative results. It had a sensitivity of 95%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 99.4%. The postoperative complications included bile leaks from the liver bed in two patients and a retained bile duct stone in one patient. If IOC had been used selectively in a traditional manner on the basis of preoperative risk factors, IOC would have been needed for 77 patients (38.5%). The total cost of LUS plus IOC for the current 200 patients was 26,256 dollars. The total estimated cost of selective IOC, if it had been performed for the 77 patients, would have been 31,416 dollars. CONCLUSIONS: Routine LUS accurately diagnosed bile duct stones and significantly reduced the need for selective IOC from a potential 38.5% to an actual 3.5% without adversely affecting the outcome of the LC or increasing the overall cost. The routine use of LUS during LC is accurate and cost effective. PMID- 17122982 TI - Low section of the rectum during laparoscopic total mesorectal excision using the Contour device. Technical report. PMID- 17122983 TI - Successful endoscopic treatment of chronic groin pain in athletes. AB - BACKGROUND: Chronic groin pain, especially in professional sportsmen, is a difficult clinical problem. METHODS: From January 1999 to August 2005, 55 professional and semiprofessional sportsmen (53 males; mean age, 25 +/- 4.5 years; range, 17-36 years) with undiagnosed chronic groin pain were followed prospectively. All the patients underwent an endoscopic total extraperitoneal (TEP) mesh placement. RESULTS: Incipient hernia was diagnosed in the study athletes: 15 on the right side (27%), 12 on the left side (22%), and 9 bilaterally (16%). In 20 patients (36%), an inguinal hernia was found: 3 direct inguinal hernias (5%) and 17 indirect hernias (31%). All the athletes returned to their normal sports level within 3 months after the operation. CONCLUSIONS: A TEP repair must be proposed to patients with prolonged groin pain unresponsive to conservative treatment. If no clear pathology is identified, reinforcement of the wall using a mesh offers good clinical results for athletes with idiopathic groin pain. PMID- 17122984 TI - Impact of intraoperative ultrasonography in laparoscopic liver surgery. AB - BACKGROUND: Laparoscopic surgery has gained growing acceptance, but this does not hold for laparoscopic surgery of the liver. This mainly includes diagnostic procedures, interstitial therapies, and treatment of liver cysts. However, the authors believe there is room for a laparoscopic approach to the liver in selected cases. METHODS: A prospective study of laparoscopic liver resections was undertaken with patients who had preoperative diagnoses of benign lesion and hepatocellular carcinoma with compensated cirrhosis. The inclusion criteria required that hepatic involvement be limited and located in the left or peripheral right segments (segments 2-6), and that the tumor be 5 cm or smaller. The location of the tumor and its transection margin were defined by laparoscopic ultrasound (LUS). RESULTS: From December 1996, 17 (5%) of 313 liver resections were included in the study. There were 5 benign lesions and 12 hepatocellular carcinomas in cirrhotic patients. The mean age of the study patients was 59 years (range, 29-79 years). The LUS evaluation identified the presence of new hepatocellular carcinoma nodules in two patients (17%). The resections included 1 bisegmentectomy, 8 segmentectomies, 3 subsegmentectomies, and 3 nonanatomic resections. The mean operative time, including laparoscopic ultrasonography, was 156 +/- 50 min (median, 150 min; range, 60-250 min), and the perioperative blood loss was 190 +/- 97 ml. There was no mortality. Conversion to laparotomy was necessary for two patients. Postoperative complications were experienced by 3 of 15 patients, all of them cirrhotics. One of the patients had a wall hematoma, and the remaining two patients had bleeding from a trocar access requiring a laparoscopic reexploration. The mean hospital stay for the whole series was 6.9 +/- 4.9 days (median, 6 days; range, 2-25 days) and 5.6 +/-1.4 days (median, 6 days; range, 2-8 days) for the 15 laparoscopic patients. CONCLUSION: Laparoscopic treatment should be considered for selected patients with benign and malignant lesions in the left lobe or frontal segments of the liver. Evaluation by LUS is indispensable to guarantee precise determination of the segmental tumor location and the relationship of the tumor to adjacent vascular or biliary structures, excluding adjacent or adjunctive new lesions. The evolution of laparoscopic hepatectomies probably will depend on the development of new techniques and instrumentations. PMID- 17122985 TI - Laparoscopic feeding jejunostomy in esophagogastric cancer. AB - BACKGROUND: Patients with esophagogastric malignancies often require nutritional supplementation in the perioperative period, especially in the setting where neoadjuvant therapy may delay tumor resection. A simple technique is described here that can be performed at the time of staging laparoscopy and that has not been described before. RESULTS: Forty-three patients treated over a 4-year period who had a laparoscopic feeding jejunostomy placed at the time of staging laparoscopy were reviewed. Of these, 35 had preoperative chemotherapy according to a modified MRC OEO2 protocol. In the period between staging and eventual resection, 32% required immediate feeding, and in 14% of those who were thought not to need feeding it later became necessary. More patients gained weight or had a rise in albumin in the group that had jejunal feeding (p < 0.05). The mean time to surgery was 10 weeks. There were no conversions to an open procedure, nor were there any laparotomies for tube-related complications. Dislodgement was recorded in 6 patients; blockage, in 4. In most of these cases a simple bedside replacement of the tube was all that was required. Mean time in the operating room for each procedure was 44 minutes. CONCLUSIONS: Laparoscopic percutaneous feeding jejunostomy is a safe and simple technique that adds little to the morbidity and cost of managing patients with esophagogastric cancers. It facilitates optimization of nutrition in the perioperative period for these patients, especially in those receiving preoperative chemotherapy. PMID- 17122986 TI - Predictive value of upper gastrointestinal studies versus clinical signs for gastrointestinal leaks after laparoscopic gastric bypass. AB - BACKGROUND: The utility of routine upper gastrointestinal (UGI) studies after laparoscopic Roux-en-Y gastric bypass (LRYGB) is a matter of great debate. Because the morbidity and mortality rates associated with an unrecognized postoperative leak are high after LRYGB, diagnosis of a postoperative leak earlier would be of benefit. Clinical signs, however, may predict the diagnosis of a postoperative leak more often. This study explored the hypothesis that UGI studies are more predictive than clinical signs for the early diagnosis of a postoperative leak after LRYGB. METHODS: All patients who underwent LRYGB at the authors' institution were included in this study. Charts were reviewed to examine immediate clinical signs (heart rate, temperature, and white blood cell count within the first 24 h), UGI studies, and clinical course. Sensitivity, specificity, positive predictive value, negative predictive value, and efficiency of clinical signs and UGI studies were calculated. RESULTS: This study included 245 patients with a 3% rate of leak. The positive and negative predictive value of UGI studies were 67% and 99%, respectively. Only an elevated white blood count had a better predictive value (100% for negative predictive value). The efficiency of UGI studies (98%) was better than that of heart rate (83%), white blood count (8%), or temperature (95%). CONCLUSIONS: According to our data, UGI studies are the most predictive of an early leak diagnosis. Clinical signs alone may not be as useful in predicting leaks early after laparoscopic gastric bypasses. Routine early postoperative UGI studies are a reasonable approach to predicting leaks after LRYGB. PMID- 17122987 TI - Is routine sentinel lymph node biopsy indicated in women undergoing contralateral prophylactic mastectomy? Magee-Womens Hospital experience. AB - INTRODUCTION: The routine use of sentinel node biopsy (SLNB) at the time of prophylactic mastectomy remains controversial. This retrospective study was undertaken to determine if SLNB is justified in patients undergoing CPM. METHODS: Between 1999 and 2004, 155 patients underwent contralateral prophylactic mastectomy (CPM) at the Magee-Womens Hospital of University of Pittsburgh Medical Center. Eighty patients (51.6%) had SLNB performed at the time of CPM. The therapeutic mastectomy and the CPM specimens were evaluated for histopathology. Goldflam's classification was used to determine the risk of malignancy in the CPM specimens. RESULTS: Pathology in the therapeutic mastectomy specimens included 105 (68%) invasive carcinomas and 50 (32%) in-situ carcinomas. Multicentricity and/or multifocality were reported in 49.7%, and 70% were estrogen receptor positive. Two invasive breast cancers and three cases of DCIS were diagnosed in 155 CPM specimens (n = 5, 3.2%). The median number of SLN identified was 2 (range 1-6) from the CPM axilla. Two patients had positive SLNB for metastatic carcinoma (n = 2/80, 2.5%) with no primary tumor identified in the prophylactic mastectomy specimen. In both patients the therapeutic mastectomy was for recurrent invasive carcinoma in patients with a prior history of axillary node dissection. Occult carcinoma was found in five prophylactic mastectomy specimens: two invasive and three DCIS. Only 1 out of the 75 patients not undergoing SLNB at the time of their initial surgery would have required axillary staging for a previously undiagnosed invasive cancer in the CPM specimen on final pathology. Of all 155 patients undergoing CPM, only 4 (2.5%) had identified final pathologic findings where axillary staging with SLNB was beneficial. There was no evidence of arm lymphedema in any patient who had undergone CPM and SLNB at a median follow-up of 24 months. CONCLUSION: Although SLNB is a minimally invasive method of axillary staging, this retrospective study does not support its routine use in patients undergoing CPM. PMID- 17122988 TI - Molecular prognostic factors in adenocarcinoma of the esophagus and gastroesophageal junction. AB - OBJECTIVE: This review describes genetic and molecular changes related to adenocarcinoma of the esophagus and gastroesophageal junction (GEJ) with emphasis on prognostic value and possibilities for targeted therapy in clinical setting. Adenocarcinoma of the esophagus or GEJ is an aggressive disease with early lymphatic and hematogenous dissemination. Molecular pathology has revealed many molecular mechanisms of disease progression, which are related to prognosis. Some of these factors can be seen as prognostic factors per se. Better knowledge of molecular bases may lead to new paradigms, improved prognostication, early diagnosis and individually tailored therapeutic options. METHODS: A review of recent English literature (1990-October 2005) concerning esophageal adenocarcinoma was performed. This review focuses on genetic and molecular changes as prognosticators of adenocarcinoma of the esophagus and GEJ. RESULTS: A bewildering number of biomarkers have been described. Many genes and molecules have prognostic impact (cyclin D1, EGFR, Her-2/Neu, APC, TGF-beta, Endoglin, CTGF, P53, Bcl-2, NF-kappaB, Cox-2, E-cadherin, beta-catenin, uPA, MMP-1,3,7,9, TIMP, T( h )1/T( h )2 balance, CRP, PTHrP). CONCLUSIONS: Adenocarcinomas of the esophagus and GEJ show multiple genetic alterations, which indicate that progression of cancer is a multistep complex process with many different alterations. Presumably, it is not one molecular factor that can predict the biological behavior of this cancer. The combination of diverse genetic alterations may better predict prognosis. In future, gene expression analysis with microarrays may reveal important prognostic information and the discovery of new genes and molecules associated with tumor progression and dissemination will enhance prognostication and offers adjuvant therapeutic options. PMID- 17122989 TI - Local/cervical block anesthesia versus general anesthesia for minimally invasive parathyroidectomy: what are the advantages? AB - BACKGROUND: Minimally invasive parathyroidectomy (MIP) under local/cervical block anesthesia (LA) is safe and effective for patients with primary hyperparathyroidism (HPT). Advantages of LA versus general anesthesia (GA) for these focused procedures have not been clearly demonstrated. METHODS: Between 3/01 and 6/04, 177 consecutive patients with primary HPT and positive localization studies underwent MIP. Seventy-three (41%) had surgery under LA while 104 (59%) had GA. Primary endpoints were IV narcotic use, anti-emetic use, nausea, vomiting, and post-operative pain. RESULTS: Patients who had parathyroidectomy under LA were older (64 +/- 2 vs. 57 +/- 2 years, P = 0.001). Cure and complication rates were identical between the two groups. Patients who had parathyroidectomy under LA required less IV narcotic pain mediation (mean morphine equivalents 11.4 +/- 1.3 mg vs. 22.5 +/- 1.1 mg; P < 0.001) compared to GA patients. The LA patients had better pain control as shown by lower post operative peak pain scores (2.9 +/- 0.3 vs. 5.0 +/- 0.4; P < 0.001) and lower overall pain scores (mean 1.9 +/- 0.2 vs. 3.1 +/- 0.2; P < 0.001). The LA group required fewer anti-emetic medications compared to the GA patients (mean 0.4 +/- 0.1 vs. 1.7 +/- 0.1 doses; P < 0.001). Fewer LA patients experienced post operative nausea (16% vs. 49%; P < 0.001), and vomiting (7% vs. 24%; P = 0.002). Length of stay was similar between the groups (0.4 +/- 0 vs. 0.3 +/- 0; P = 0.22). CONCLUSIONS: In this study the choice of anesthesia did not affect surgical cure rate, morbidity, or length of stay. LA was associated with significantly lower post-operative pain, nausea, and vomiting. LA appears to offer specific advantages more than GA for patients undergoing MIP. PMID- 17122990 TI - Radioguided parathyroidectomy in patients with familial hyperparathyroidism. AB - BACKGROUND: In patients with sporadic hyperparathyroidism (HPT), radioguided parathyroidectomy (RGP) has been shown to facilitate intraoperative localization of parathyroid glands, reduced operative time, and improve patient outcomes. No studies have focused on the role of RGP in patients with familial HPT. METHODS: Between 3/01 and 6/05, 419 patients underwent RGP. Nineteen had familial HPT, including 12 with Multiple Endocrine Neoplasia (MEN), and 94 had sporadic HPT with parathyroid hyperplasia. All patients were injected with sestamibi pre operatively and a gamma probe was used intraoperatively. Radiotracer counts were recorded prospectively. RESULTS: In patients with familial HPT, the gamma probe detected all abnormal parathyroid glands with a mean in vivo radiotracer count of 157 +/- 9% above background. Importantly, 5 patients (25%) had ectopic parathyroid glands localized by the probe in the thymus, thyroid and retroesophageal region. All resected hyperplastic parathyroid glands had ex vivo counts > 20%. All patients were cured after surgery with mean calcium and parathyroid hormone levels of 9.4 +/- 0.1 mg/dl and 31 +/- 7 pg/ml, respectively, and a mean hospital stay of 0.7 +/- 0.1 days. In comparing the 2 groups, while patients with familial HPT had lower pre-operative parathyroid hormone levels, the ex vivo radiotracer counts were significantly higher. CONCLUSION: RGP in patients with familial HPT is technically feasible and perhaps more sensitive than in patients with sporadic hyperplastic disease. The gamma probe efficiently localized all parathyroid glands including those in ectopic locations, and resulted in high cure rates and short hospital stays. RGP is a viable and useful technique in patients with familial HPT. PMID- 17122991 TI - Advanced extremity soft tissue sarcoma: prognostic effect of isolated limb perfusion in a series of 88 patients treated at a single institution. AB - PURPOSE: To explore the prognostic impact of isolated limb perfusion (ILP) in locally advanced extremity soft tissue sarcomas (ESTS). METHODS: From August 1982 to April 2005, 1,119 patients affected by ESTS (girdle excluded) were observed and treated at our institution. Eighty-eight (7.9%) were judged non-resectable or locally advanced and underwent ILP. Thirty-seven patients received antiblastic alone (non-TNF-ILP) while 51 had anti-blastic + recombinant-tumor necrosis factor alpha (TNF-ILP). Local disease-free survival (LDFS) was calculated by the Kaplan Maier method and was reported separately in the two subgroups. RESULTS: Limb salvage was achieved in 83% (73/88) of the patients. The observed overall (complete + partial) response rate was 59%. In the TNF-ILP group a complete response (CR) was achieved in 21 (41%) patients, while in the non-TNF ILP group a CR was obtained in seven (19%) cases (P < 0.05). Patients with in-transit metastases (epithelioid sarcomas and clear cell sarcomas) had a significantly worse long-term outcome (LDFS at 5 years was 40.9 vs 67.3%, P < 0.05). A trend towards a better LDFS at 5 years could be observed in the patients receiving TNF (63.6 vs 57.1%) and post-operative radiation therapy (RT) (79.3 vs 55.4%). CONCLUSIONS: Isolated limb perfusion is an active treatment. By adding TNF a better local control seems to be obtained, possibly due to a higher rate of CR. It should therefore be considered as a valid option for patients affected by limb threatening STS, save for in-transit metastases from epithelioid and clear cell sarcoma. Post-operative RT should always be considered. PMID- 17122992 TI - Timing of adjuvant radioimmunotherapy after cytoreductive surgery in experimental peritoneal carcinomatosis of colorectal origin. AB - BACKGROUND: Treatment of patients with peritoneal carcinomatosis (PC) of colorectal cancer (CRC) includes cytoreductive surgery (CS) in combination with (hyperthermic) intraperitoneal chemotherapy (HIPEC), resulting in a limited survival benefit with high morbidity and mortality rates. Radioimmunotherapy (RIT) as adjuvant therapy after CS of CRC has been shown to prolong survival in preclinical studies. However, the optimal setting of RIT remains to be determined. METHODS: PC was induced by intraperitoneal inoculation of CC-531 colon carcinoma cells in Wag/Rij rats. Animals were subjected to exploratory laparotomy (Sham), CS only or CS + RIT at different time points after surgery. RIT consisted of 55 MBq lutetium-177-labelled anti-CC531 antibody MG1 (183 mug). The primary endpoint was survival. RESULTS: Cytoreductive surgery with or without RIT was well tolerated. Median survival of animals in the Sham and CS group was 29 days and 39 days, respectively (P < 0.04). Compared to CS alone, median survival of rats after adjuvant RIT was 77 days (P < 0.0001), 52 days (P < 0.0001) and 45 days (P < 0.0001) when given directly, 4 and 14 days after surgery, respectively. CONCLUSION: The efficacy of adjuvant RIT after CS for the treatment of PC of colonic origin decreases when the administration of the radiolabelled MAbs is postponed. This study shows that adjuvant RIT should be given as early as possible after surgery. PMID- 17122993 TI - Usefulness of texture analysis for computerized classification of breast lesions on mammograms. AB - This work presents the usefulness of texture features in the classification of breast lesions in 5,518 images of regions of interest, which were obtained from the Digital Database for Screening Mammography that included microcalcifications, masses, and normal cases. Sixteen texture features were used, i.e., 13 were based on the spatial gray-level dependence matrix and 3 on the wavelet transform. The nonparametric K-NN classifier was used in the classification stage. The results obtained from receiver operating characteristic analysis indicated that the texture features can be used for separating normal regions and lesions with masses and microcalcifications, yielding the area under the curve (AUC) values of 0.957 and 0.859, respectively. However, the texture features were not very effective for distinguishing between malignant and benign lesions because the AUC was 0.617 for masses and 0.607 for microcalcifications. The study showed that the texture features can be used for the detection of suspicious regions in mammograms. PMID- 17122996 TI - Reconstituted botulinum type A neurotoxin: clinical efficacy after long-term freezing before use. AB - Botulinum neurotoxin type A (BoNT/A) is injected into muscles of facial expression for treatment of facial wrinkles. Manufacturers' recommendations state that the product should be used within 4 h of reconstitution to prevent loss of effectiveness. A prospective study was undertaken to evaluate the effectiveness of reconstituted BoNT/A when preserved in a frozen state for up to 6 months, as compared with nonfrozen BoNT/A used within 4 h after reconstitution. Treatment of 118 sites in 80 patients was performed and evaluated for aesthetic efficacy. Excellent results were observed in 94.8% of sites treated with nonfrozen BoNT/A and in 93.3% of the sites treated with frozen and thawed BoNT/A (p = 1). Minor side effects were rare and transient in both groups. In contrast to common belief, reconstituted BoNT/A may be frozen, thawed, and injected without losing its potency for up to 6 months, with efficacy equivalent to that of freshly prepared BoNT/A. PMID- 17122997 TI - Initial screening of contaminated land: a comparison of US and Swedish methods. AB - Preliminary surveys are used to prioritize between contaminated sites to select those to be investigated more thoroughly. The data-gathering steps are almost identical between countries; however, the assessment procedures differ significantly. In this study, we have investigated 21 contaminated sites assessed as belonging to the high-risk or the very high-risk class using the Swedish Methods for Inventories of Contaminated Sites (MICS). We then applied the US Preliminary Assessment (PA) method to the same sites and compared the results and conclusions from the two screening procedures. In both cases, all sites were recommended for further investigation and the two approaches seem to corroborate one another; however, the PA assessment scores and the preliminary MICS classifications did not correlate. The results obtained with the PA method were easier to explain than the final MICS classification. The PA method also seems more transparent and easier to standardize, although objections could be made regarding the weighting scheme, because the outcome in this study was entirely dependent on the surface exposure pathway. However, to examine this in greater detail, it is necessary to include sites with less contamination: The importance of preliminary surveys in the overall risk management process gives a strong motivation for such an evaluation. Generally, the lack of research and scientific support for the various assessment procedures in use suggests that there is a need for method development, standardization, and validation. PMID- 17122998 TI - Linkages between nutrients and assemblages of macroinvertebrates and fish in wadeable streams: implication to nutrient criteria development. AB - We sampled 240 wadeable streams across Wisconsin for different forms of phosphorus and nitrogen, and assemblages of macroinvertebrates and fish to (1) examine how macroinvertebrate and fish measures correlated with the nutrients; (2) quantify relationships between key biological measures and nutrient forms to identify potential threshold levels of nutrients to support nutrient criteria development; and (3) evaluate the importance of nutrients in influencing biological assemblages relative to other physicochemical factors at different spatial scales. Twenty-three of the 35 fish and 18 of the 26 macroinvertebrate measures significantly correlated (P < 0.05) with at least one nutrient measure. Percentages of carnivorous, intolerant, and omnivorous fishes, index of biotic integrity, and salmonid abundance were fish measures correlated with the most nutrient measures and had the highest correlation coefficients. Percentages of Ephemeroptera-Plecoptera-Trichoptera individuals and taxa, Hilsenhoff biotic index, and mean tolerance value were macroinvertebrate measures that most strongly correlated with the most nutrient measures. Selected biological measures showed clear trends toward degradation as concentrations of phosphorus and nitrogen increased, and some measures showed clear thresholds where biological measures changed drastically with small changes in nutrient concentrations. Our selected environmental factors explained 54% of the variation in the fish assemblages. Of this explained variance, 46% was attributed to catchment and instream habitat, 15% to nutrients, 3% to other water quality measures, and 36% to the interactions among all the environmental variables. Selected environmental factors explained 53% of the variation in macroinvertebrate assemblages. Of this explained variance, 42% was attributed to catchment and instream habitat, 22% to nutrients, 5% to other water quality measures, and 32% to the interactions among all the environmental variables. PMID- 17122999 TI - Assessing potential removal of low-head dams in urban settings: an example from the Ottawa River, NW Ohio. AB - This is a study of the scientific component of an effort to restore an urban river by removing a low-head dam. The Secor Dam is owned by a local government entity near Toledo, Ohio. The proposed removal of the last structure impeding flow on the Ottawa River has broad appeal, but the owner is concerned about liability issues, particularly potential changes to the flood regime, the presence of contaminated sediments behind the dam, and possible downstream transport of reservoir sediments. Assessing sediment contamination involved sediment sampling and analysis of trace metals and organic contaminants. Forecasting sediment transport involved field methods to determine the volume and textural properties of reservoir and upstream sediment and calculations to determine the fate of reservoir sediments. Forecasting changes in the flood regime involved HEC-RAS hydrological models to determine before and after dam removal flood scenarios using LiDAR data imported into an ArcGIS database. The resulting assessment found potential sediment contamination to be minor, and modeling showed that the removal of the dam would have minimal impacts on sediment transport and flood hazards. Based on the assessment, the removal of the dam has been approved by its owners. PMID- 17123000 TI - Application of a karst disturbance index in Hillsborough County, Florida. AB - Hillsborough County, Florida, is a karst region that is heavily urbanized, yet no study has been undertaken measuring the degree of human disturbance. Van Beynen and Townsend (2005) created a hierarchical and standardized disturbance index specifically designed for karst environments. To address the problem of determining human disturbance in the county, the above index was successfully applied and it was found that Hillsborough was highly disturbed (disturbance score of 0.69 of 1.0) because of its predominant urban and rural land use. Furthermore, the application of the index allowed for its refinement and the highlighting of environmental aspects in need of remediation such as soil compaction, deforestation, disturbance of archaeological sites, and the expanding urban footprint. Several minor issues arose during the application: the need for broader indicator descriptions that encompass a variety of scenarios, the need for a revised water quality indicator, inadequate data on sinkholes, and a lack of data for species richness and species population density. The utility of the index to resource managers arises from emphasizing certain areas of the environment that require immediate attention and determining temporal changes in environmental quality. Future application of the index requires potential retooling of the biota indicators, tightening of scoring descriptions for certain indicators, and further examination of the scale at which the index can be applied. PMID- 17123001 TI - Remote sensing and GIS techniques for selecting a sustainable scenario for Lake Koronia, Greece. AB - During recent decades, Lake Koronia has undergone severe degradation as a result of human activities around the lake and throughout the basin. Surface and groundwater abstraction and pollution from agricultural, industrial, and municipal sources are the major sources of degradation. Planning a restoration project was hampered by lack of sufficient data, with gaps evident in both spatial and temporal dimensions. This study emphasized various remote sensing and geographic information system techniques, such as digital image processing and geographic overlay, to fill gaps using satellite imagery and other spatial environmental, hydrological, and hydrogeological data in the process of planning the restoration of Lake Koronia, following Ramsar guidelines. Current and historical remote sensing data were used to assess the current status and level of degradation, set constraints and define the ideotype for the restoration, and, finally, define and select the best restoration scenario. PMID- 17123002 TI - It's all in the numbers: acreage tallies and environmental program evaluation. AB - Increasingly, performance measurement is being used to hold federal agencies accountable, represent environmental progress, and evaluate the effectiveness of environmental programs. The need to track measurable outputs has created a tendency to present programmatic progress solely by quantifiable data, despite the inherent complexity of natural resource management. Wetlands and fire management programs are two specific environmental arenas that have come to overemphasize the tracking of acreage numbers to validate existing policy direction. In both of these arenas, we find the definition and categorization of "countable" acres to be inconsistent and unreliable. We explore this systemic flaw for both wetlands and fire programs and describe its implications for environmental policy and natural resource management more broadly. We conclude with recommendations for improved performance measurement in these arenas. PMID- 17123003 TI - Integrating human impacts and ecological integrity into a risk-based protocol for conservation planning. AB - Conservation planning aims to protect biodiversity by sustainng the natural physical, chemical, and biological processes within representative ecosystems. Often data to measure these components are inadequate or unavailable. The impact of human activities on ecosystem processes complicates integrity assessments and might alter ecosystem organization at multiple spatial scales. Freshwater conservation targets, such as populations and communities, are influenced by both intrinsic aquatic properties and the surrounding landscape, and locally collected data might not accurately reflect potential impacts. We suggest that changes in five major biotic drivers-energy sources, physical habitat, flow regime, water quality, and biotic interactions-might be used as surrogates to inform conservation planners of the ecological integrity of freshwater ecosystems. Threats to freshwater systems might be evaluated based on their impact to these drivers to provide an overview of potential risk to conservation targets. We developed a risk-based protocol, the Ecological Risk Index (ERI), to identify watersheds with least/most risk to conservation targets. Our protocol combines risk-based components, specifically the frequency and severity of human-induced stressors, with biotic drivers and mappable land- and water-use data to provide a summary of relative risk to watersheds. We illustrate application of our protocol with a case study of the upper Tennessee River basin, USA. Differences in risk patterns among the major drainages in the basin reflect dominant land uses, such as mining and agriculture. A principal components analysis showed that localized, moderately severe threats accounted for most of the threat composition differences among our watersheds. We also found that the relative importance of threats is sensitive to the spatial grain of the analysis. Our case study demonstrates that the ERI is useful for evaluating the frequency and severity of ecosystem-wide risk, which can inform local and regional conservation planning. PMID- 17123004 TI - Development of an ecologic marine classification in the new zealand region. AB - We describe here the development of an ecosystem classification designed to underpin the conservation management of marine environments in the New Zealand region. The classification was defined using multivariate classification using explicit environmental layers chosen for their role in driving spatial variation in biologic patterns: depth, mean annual solar radiation, winter sea surface temperature, annual amplitude of sea surface temperature, spatial gradient of sea surface temperature, summer sea surface temperature anomaly, mean wave-induced orbital velocity at the seabed, tidal current velocity, and seabed slope. All variables were derived as gridded data layers at a resolution of 1 km. Variables were selected by assessing their degree of correlation with biologic distributions using separate data sets for demersal fish, benthic invertebrates, and chlorophyll-a. We developed a tuning procedure based on the Mantel test to refine the classification's discrimination of variation in biologic character. This was achieved by increasing the weighting of variables that play a dominant role and/or by transforming variables where this increased their correlation with biologic differences. We assessed the classification's ability to discriminate biologic variation using analysis of similarity. This indicated that the discrimination of biologic differences generally increased with increasing classification detail and varied for different taxonomic groups. Advantages of using a numeric approach compared with geographic-based (regionalisation) approaches include better representation of spatial patterns of variation and the ability to apply the classification at widely varying levels of detail. We expect this classification to provide a useful framework for a range of management applications, including providing frameworks for environmental monitoring and reporting and identifying representative areas for conservation. PMID- 17123005 TI - Modulatory effects of fresh garlic extract on chrysotile asbestos induced genotoxicity: an in vitro study. PMID- 17123006 TI - Effect of temporary closure of the mouth of an estuary on the benthic macroinfauna: Lenga-Chile, a case study. PMID- 17123007 TI - Polycyclic aromatic hydrocarbons in fruits and vegetables grown in the Czech republic. PMID- 17123008 TI - Distribution and biodegradation of bisphenol A in water hyacinth. PMID- 17123009 TI - Runoff of aerially applied phthalide from paddy fields. PMID- 17123010 TI - Methomyl residue on Chinese cabbage grown under greenhouse conditions. PMID- 17123011 TI - Dissipation of malathion on Fenugreek (Trigonella foenum-graceum L.). PMID- 17123012 TI - Bioaccumulation of the antifouling paint booster biocide Irgarol 1051 by the green alga Tetraselmis suecica. PMID- 17123013 TI - Extinction coefficients and dissolved organic carbon content in freshwater in Kenya. PMID- 17123014 TI - Acute toxicity of permethrin to four populations of ovigerous grass shrimp, Palaemonetes pugio Holthuis. PMID- 17123015 TI - Toxic effects of lead on the liver and gills of Oncorhynchus mykiss WALBAUM 1792. PMID- 17123016 TI - CYP1A expression in Mullus barbatus and Merluccius merluccius from the Adriatic Sea in Serbia and Montenegro. PMID- 17123017 TI - Effects of toxic and nontoxic Microcystis aeruginosa on survival, population increase, and feeding of two small cladocerans. PMID- 17123018 TI - Metal content in Ulva lactuca (Linnaeus) from Navachiste Bay (southeast Gulf of California) Sinaloa, Mexico. PMID- 17123019 TI - Accumulation of nutrients and metal ions by two mung bean [Vigna radiata (L.) Wilczek] cultivars treated with copper and lead. PMID- 17123020 TI - Trace metal concentrations in wild and cultured Australian short-finned eel (Anguilla australis Richardson). PMID- 17123021 TI - Mercury, cadmium, selenium, and seven other elements in the muscle, renal, and hepatic tissue of harbor seals (Phoca vitulina) from Newfoundland and Labrador, Canada. PMID- 17123022 TI - Heavy metals from agricultural soils from Cameron Highlands, Pahang, and Cheras, Kuala Lumpur, Malaysia. PMID- 17123023 TI - Toxicity and trace metal concentrations of sediments from Lake Maryut, Alexandria, Egypt. PMID- 17123024 TI - PM10 metal distribution in an industrialized city. PMID- 17123025 TI - The recent evolution of pentachlorophenol (PCP)-4-monooxygenase (PcpB) and associated pathways for bacterial degradation of PCP. AB - Man-made polychlorinated phenols such as pentachlorophenol (PCP) have been used extensively since the 1920s as preservatives to prevent fungal attack on wood. During this time, they have become serious environmental contaminants. Despite the recent introduction of PCP in the environment on an evolutionary time scale, PCP-degrading bacteria are present in soils worldwide. The initial enzyme in the PCP catabolic pathway of numerous sphingomonads, PCP-4-monooxygenase (PcpB), catalyzes the para-hydroxylation of PCP to tetrachlorohydroquinone and is encoded by the pcpB gene. This review examines the literature concerning pcpB and supports the suggestion that pcpB/PcpB should be considered a model system for the study of recent evolution of catabolic pathways among bacteria that degrade xenobiotic molecules introduced into the environment during the recent past. PMID- 17123027 TI - Reaction between alkyl isocyanides and isopropylidene Meldrum's acid in the presence of bidentate nucleophiles. AB - The reaction between alkyl isocyanides and isopropylidene Meldrum's acid in the presence of 1,2-ethanediol leads to N (1)-(alkyl)-2-(5,7-dioxo-1,4-dioxepane-6 yl)-2-methylpropanamides. 1,3-Propanediol or 1,4-butanediol produce hydroxyalkyl 1-(tert-butyl)-4,4-dimethyl-2,5-dioxo-3-pyrrolidinecarboxylates. When the reaction was performed in the presence of catechol, bis(2-hydroxyphenyl) 2-[2 (tert-butylamino)-1,1-dimethyl-2-oxoethyl]malonate was obtained. 2-Aminophenols react with alkyl isocyanides in the presence of isopropylidene Meldrum's acid to produce 1-alkyl-N (3)-(2-hydroxyaryl)-4,4-dimethyl-2,5-dioxo-3 pyrrolidinecarboxamides in good yields. PMID- 17123028 TI - Combination pharmacotherapy in hypertension. AB - Combination pharmacotherapy with two or more drugs is required in order to reach the currently recommended blood pressure goals in the majority of hypertensive patients, particularly those with a goal of <130/80 mm Hg. Further to the potentiation of the antihypertensive effects, benefits of combination therapy include the potential of fewer adverse affects and of improvement of patients' compliance. Current guidelines recommend that combination pharmacotherapy might also be considered as initial treatment in patients with significant elevation of blood pressure and evidence of complications. Several effective and well tolerated antihypertensive drug classes available today offer multiple options for combination therapy. The choice of antihypertensive agents should be made on the basis of current recommendations regarding first line drugs and compelling indications. Specific drug combinations might have additional beneficial or detrimental long-term metabolic effects, beyond their effects on blood pressure. However, more outcome data comparing antihypertensive drug combinations are required. The implementation of an intensive up-titration treatment strategy, together with a systematic use of full doses of multiple drug combinations, is expected to achieve optimal blood pressure control in the vast majority of hypertensive patients. PMID- 17123029 TI - Serologic testing for symptomatic coccidioidomycosis in immunocompetent and immunosuppressed hosts. AB - Serologic studies are an important diagnostic tool in the clinical evaluation and follow-up of persons with coccidioidomycosis. Numerous types of serologic tests are available, including immunodiffusion, enzyme immunoassay, and complement fixation. We conducted a retrospective review of the results of 1,797 serologic tests spanning 12 months from the onset of coccidioidomycosis in 298 immunocompetent and 62 immunosuppressed persons with symptomatic infection. Using the onset of symptoms as a reference point, we plotted the positive or negative serologic results over time for both groups. Compared with the immunocompetent group, immunosuppressed persons had lower rates of seropositivity for every type of test during the first year after onset of symptoms for coccidioidomycosis, although many results did not achieve statistical significance. Combining the results of these tests increased the sensitivity of the serologic evaluation in immunocompromised patients. Immunosuppressed persons have the ability to mount a serologic response to coccidioidomycosis, but in some circumstances, multiple methods may be required to improve detection. PMID- 17123030 TI - Serological detection of antibodies against Paracoccidioides brasiliensis in dogs with leishmaniasis. AB - The aim of this study was to detect antibodies against Paracoccidioides brasiliensis in dogs seropositive and seronegative for leishmaniasis. Sera from 836 dogs (449 positive and 387 negative to leishmaniasis) were analysed by ELISA and the immunodiffusion test using gp43 and exoantigen, respectively. The analysis of the 836 serum samples by ELISA and the immunodiffusion test showed a positivity of 67.8 % and 7.3%, respectively, for P. brasiliensis infection. The dogs positive to leishmaniasis showed a higher reactivity to gp43 (79.9%) and exoantigen (12.7%) than the negative ones (54.0% and 1.0%, respectively). The higher reactivity to P. brasiliensis antigens may be due to cross-reactivity or a co-infection of dogs by Leishmania and P. brasiliensis. The lower correlation (0.187) observed between reactivity to gp43 and Leishmania antigen reinforces the latter hypothesis. PMID- 17123031 TI - Role of mast cells as IL10 producing cells in paracoccidioidomycosis skin lesions. AB - Recent works have demonstrated that mast cells may have an important role in immunologic reactions and inflammation once they synthesize and secrete many cytokines including IL4, IL5, IL6 and TNF-alpha. We have conducted research in order to verify if mast cells would participate in the local inflammatory immune response against Paracoccidioides brasiliensis in skin lesions characterized by a Th2 pattern of cytokines. Fifty-nine skin biopsies with previous histopathological diagnosis of paracoccidioidomycosis and immunohistochemical characterization of cytokines present in the inflammatory infiltrate were classified in three groups: group 1 (G1), with compact granuloma and a Th1 pattern of cytokines; group 2 (G2), with loose granuloma and a Th2 pattern of cytokines; group 3 (G3), both kind of granuloma in the same lesion, characterized by cytokines from Th1 and Th2 patterns. Ten biopsies from normal skin were used as control group. Mast cells were visualized and quantified by a toluidine blue/HCl staining and a double immunostaining was performed to detect a co localization of mast cells and IL10. G2 presented an increased number of mast cells when compared to G1, G3 and control group and we frequently could find mast cells expressing IL10 in G2. The data obtained suggest that mast cells participate in the immune response against P. brasiliensis in skin lesions with loose granuloma and a Th2 pattern of cytokines. Considering these results, mast cells could constitute a source of IL10, contributing to a non-effective response against fungal antigens. PMID- 17123032 TI - Characterization of an extracellular endopolygalacturonase from the saprobe Mucor ramosissimus Samutsevitsch and its action as trigger of defensive response in tropical plants. AB - In recent years, interest in the ability of non-pathogenic microorganisms to induce resistance in plants has grown, particularly with respect to their use as environmentally safe controllers of plant disease. In this study, we investigated the capacity of Mucor ramosissimus Samutsevitsch to release pectinases able to degrade cell walls of Palicourea marcgravii St. Hil., a tropical forest native Rubiaceae on which the spores of this saprobic fungus have been found. The fungus was grown in liquid culture medium containing pectin as the sole carbon source and filtrates were analyzed for pectinase activity. An endopolygalacturonase was partially purified by ion exchange chromatography, gel filtration, and preparative isoelectrofocusing, and characterized. This enzyme was more active upon pectic substrates with a low degree of methyl esterification. The products of hydrolysis of different pectic substrates (including pectin from P. marcgravii) by the action of this endopolygalacturonase elicited to different extents the phytoalexin production in soybean cotyledons. Also, the enzyme itself and the products of its action on the pectic fraction of P. marcgravii elicited the production of defensive compounds in the leaves of the plant. These results suggest that, besides the role in recycling organic matter, saprobes may also play an important role in the induction of defensive mechanisms in wild plants by enhancing their non-specific resistance against pathogens. Furthermore, they set the stage for future studies on the role of saprobic fungi in inducing resistance of host plants to pathogens. PMID- 17123033 TI - Impact on growth and aflatoxin B1 accumulation by Kluyveromyces isolates at different water activity conditions. AB - This study showed the impact on germination, mycelial growth and aflatoxin B(1) accumulation when interacting Aspergillus aflatoxigenic strains with Kluyveromyces isolates and the effect of water activity on this relationship. Isolates Y(14) and Y(16) reduced the percentage of germination of all Aspergillus strains and decrease germ tube elongation rate at majority of water activity assayed. Similarly they produced an increase of germination lag phase and lag phase of growth beside decreased growth rate of all Aspergillus strains. At water activities 0.994, 0.982, 0.955 and 0.937, no aflatoxins were produced in paired cultures with isolates Y(25,) Y(22), Y(16), and Y(14), and Kluyveromyces isolates Y(14) and Y(16) impact both growth and aflatoxin accumulation at wide range of water activity. PMID- 17123034 TI - Mycobiota in poultry feeds and natural occurrence of aflatoxins, fumonisins and zearalenone in the Rio de Janeiro State, Brazil. AB - The intake of mycotoxin-contaminated feeds can lead to nutrient losses and may have adverse effects on animal health and on productivity. The aims of this study were (1) to determine the mycobiota present in poultry feed samples, and (2) to evaluate the natural occurrence of aflatoxin B(1), fumonisin B(1) and zearalenone. Fungal counts were similar between all culture media tested (10(3 )CFU g(-1)). The most frequent genus isolated was Penicillium spp. (41.26%) followed by Aspergillus spp. (33.33%) and Fusarium spp. (20.63%). High precision liquid chromatography was applied to quantify aflatoxin B(1) and fumonisin B(1). Thin layer chromatography was used to determine zearalenone levels. Aflatoxin B(1 )values ranged between 1.2 and 17.5 microg kg(-1). Fumonisin B(1) levels ranged between 1.5 and 5.5 microg g(-1). Zearalenone levels ranged between 0.1 and 7 microg g(-1). The present study shows the simultaneous occurrence of two carcinogenic mycotoxins, aflatoxin B(1) and fumonisin B(1), together with another Fusarium mycotoxin (zearalenone) in feed intended for poultry consumption. Many samples contained AFB(1 )levels near the permissible maximum and it could affect young animals. A synergistic toxic response is possible in animals under simultaneous exposure. PMID- 17123035 TI - Isolation of Cryptococcus laurentii from Canada Goose guano in rural upstate New York. AB - Cryptococcus neoformans and Cryptococcus gattii are etiologic agents of cryptococcal pneumonia and meningitis, potentially lethal syndromes associated with AIDS. A related species, Cryptococcus laurentii, has recently been implicated in several cases of human disease. Guano from Canada Goose (Branta canadensis), an organism that lives closely beside man and inhabits recreational space in rural and suburban areas, might be a significant environmental reservoir of Cryptococcus organisms in non-urban areas. Cryptococcal organisms were isolated from Canada Goose guano from a site in rural northern New York, with identification based upon colony and microscopic morphology, ability to metabolize L: -Dopa to melanin, and positive reaction with a commercial anti cryptococcal capsular polysaccharide latex bead agglutination test. DNA sequences from five positive isolates were identical to each other, and identical to the ITS1-5.8S-ITS2 sequences of C. laurentii strain CBS7140 (Accession AY315665) across a 511 bp sequence. All five isolates of C. laurentii possess three of the known virulence factors common to cryptococcal organisms that cause human disease: capsule, ability to grow at 37 degrees C, and laccase activity. PMID- 17123036 TI - Isolation of Paecilomyces lilacinus (Thom) Samson (Ascomycota: Hypocreales) from the Chagas disease vector, Triatoma infestans Klug (Hemiptera: Reduviidae) in an endemic area in Argentina. AB - A survey for entomopathogenic fungi of the Chagas disease vector Triatoma infestans was conducted in two provinces of Argentina from March-December 2003. Field-collected insects that died in the laboratory were individually maintained in moist chamber and incubated at 22 degrees C. Triatominae adults infected with the fungus Paecilomyces lilacinus were found at El Quebracho (27 degrees 34'S-64 degrees 31'W), Santiago del Estero province, Argentina, in December 2003. Paecilomyces lilacinus was cultured and isolated from infected insects in SDAY, PYG and MEA media. Pathogenicity tests were conducted and positive results were recorded. The median survival time (MST) of T. infestans exposed to a P. lilacinus conidial suspension was 12.8 days, and 100% mortality occurred at 30 days post-treatment. This is the first record of natural infection caused by P. lilacinus in T. infestans in the world. PMID- 17123039 TI - [Abstracts of the 34th Annual Meeting of the Austrian Society of Diabetes, 16-18 November 2006, Innsbruck, Austria]. PMID- 17123038 TI - Intracranial pressure levels and single wave amplitudes, Glasgow Coma Score and Glasgow Outcome Score after subarachnoid haemorrhage. AB - OBJECT: To relate intracranial pressure (ICP) levels and single ICP wave amplitudes to the acute clinical state (Glasgow Coma Score, GCS) and final clinical outcome (Glasgow Outcome Score, GOS) in patients with subarachnoid haemorrhage (SAH). METHODS: Twenty-seven consecutive patients with severe SAH had their ICP and arterial blood pressure (ABP) continuously monitored during days 1 6 after SAH. The acute clinical state could be assessed in 11 non-sedated cases using the Glasgow Coma Scale, while outcome was assessed in all cases after 6 months using the Glasgow Outcome Scale. The ICP/ABP recordings were stored as raw data files and analyzed retrospectively. For every consecutive 6 seconds time window, mean ICP, mean cerebral perfusion pressure (CPP) and the mean ICP wave amplitude were computed. RESULTS: The GCS during days 1-6 after SAH was significantly related to the mean ICP wave amplitude, but not to the mean ICP or mean CPP. There was also a strong relationship between the mean ICP wave amplitude and GOS 6 months after SAH, with mean ICP wave amplitudes being significantly lower in those with moderate disability/good recovery, as compared with those with severe disability and death. Mean ICP was significantly higher in those who died than in the group with moderate disability/good recovery whereas mean CPP was not different between outcome groups. CONCLUSIONS: In this small patient group the mean ICP wave amplitude during days 1-6 after SAH was related to the acute clinical state (GCS) as well as to the clinical outcome (GOS) 6 months after SAH. Similar relationships were not found for mean ICP or the mean CPP, except for a higher mean ICP in those who died than in those with moderate disability/good recovery. PMID- 17123040 TI - [Emergency ward management of traumatic head injury in children]. AB - BACKGROUND: The controversial situation relating to assessment and management of the traumatic head injury (THI) in children inspired us to study our own patient pool. The aims were to find a significant correlation between skull fracture or clinical symptom and intracranial lesion as well as to determine the importance of each radiological diagnostic method in the initial management of the pediatric THI. PATIENTS AND METHODS: In 1 year 1,637 children had been treated in the emergency room of pediatric surgery with the diagnosis of THI. Age, sex, injury pattern, symptoms, radiological diagnostic methods, diagnosis, and clinical follow-up had been registered. RESULTS: A significant correlation between skull fracture or clinical symptom and the intracranial injury in children could not be found, but risk factors exist. Cranial computed tomography is the imaging method of choice. X-ray, ultrasound, and MRI of the head are reserved for a few indications. CONCLUSION: A management plan for pediatric head and brain injury in the emergency room based on our own and published international results is introduced. PMID- 17123041 TI - [The AO/ASIF Flexnail : A flexible intramedullary nail for the treatment of humeral shaft fractures]. AB - BACKGROUND: The use of conventional implants for intramedullary nailing of humeral shaft fractures is associated with specific difficulties. During antegrade implantation structures of the rotator cuff can be affected leading to a reduced functional result of the shoulder. If the nail is implanted in a retrograde manner problems arise due to a relatively large hole close to or within the olecranon fossa, which is necessary for insertion of the nail. Supracondylar fractures as well as persistent elbow pain and loss of function are reported in the literature. To overcome these disadvantages a flexible nail has been developed that can be stiffened and locked after implantation. METHOD: Between October 2000 and February 2002, 34 patients were treated with the flexible nail at our institution; 29 were available for follow-up. Fracture healing was documented on radiographs and clinical outcome was evaluated with use of the Constant as well as the Kwasny score. RESULTS: Median duration until fracture consolidation was 10 weeks. In two patients fracture union was not achieved within the follow-up period. The median outcome measured with the Constant score was 93 points and 2.5 with the Kwasny score. Both values correspond to a very good functional outcome. CONCLUSION: We conclude that the flexible humeral nail is an excellent treatment option for humeral shaft fractures. Damage to the rotator cuff and the distal humerus can be avoided due to its unique flexible construction, improving the functional outcome of intramedullary nailing for the treatment of humeral shaft fractures. PMID- 17123042 TI - [Electronic documentation of injuries of the hand with a semantic network: effective and efficient methods for the documentation of clinical and administrative processes]. AB - BACKGROUND: An efficient medical documentation is mandatory for a trauma-oriented department in the DRG environment. Besides the continuously increasing clinical/administrative demands, the additional documentation for quality assurance, clinical studies, and research requires additional efforts. Standard solutions are only partially effective. Especially in hand surgery there is a high demand for sophisticated clinical documentation, represented by a wide variety of classifications in diagnosis and therapy. The standard documentation tools lack accuracy. The development of a software tool that defines administrative/business processes and simultaneously generates clinical and administrative information was the goal of this project. METHODS AND RESULTS: With a standard medical terminology, an innovative semantic network, and a completely new graphical user interface, it was possible to develop and introduce a software program specifically adjusted for hand surgery. This program facilitated for the first time a single-stage acquisition of clinically relevant scientific data and the simultaneous generation of DRG, quality assurance, and administrative data relevant for the hospital's revenues. CONCLUSIONS: The newly developed software tool is a step forward into a new dimension of medical software, obviating the need for multi/documentation and significantly improving the quality of clinically relevant medical data. PMID- 17123043 TI - [Urogenital injuries accompanying pelvic ring fractures]. AB - BACKGROUND: Follow-up of patients with pelvic ring fractures and associated injuries of the lower urogenital tract was performed from January 2000 to October 2004. Analysis focused on incidence, fracture type, type of urogenital injury, associated intrapelvic lesions, mortality, and urologic outcome. METHOD AND RESULTS: The retrospective study included 18 of 111 patients (16.2%). Nine patients had a rupture of the urethra, six a rupture of the bladder, three a rupture of the penile root, and two a gonadal defect. The type of the pelvic ring fracture according to the AO classification was type A in 1, type B in 6, and type C in 11 cases. Fifteen patients (83.3%) were followed up clinically for a mean duration of 26 months (range: 12-66 months) after trauma. Seven patients were asymptomatic concerning the urogenital injury, five had erectile dysfunction, two had urethral stenosis, in one case associated with incontinence, and one patient with bilateral defect of the testicles was under hormone substitution therapy. CONCLUSION: Urogenital injuries, often associated with intrapelvic lesions in so-called complex pelvic trauma, are typical for high grade pelvic ring fractures and have an essential prognostic value for the patient's morbidity and quality of life. PMID- 17123044 TI - [Implantation of total prosthesis for omarthritis]. PMID- 17123045 TI - [The spontaneous spinal epidural haematoma]. AB - Spontaneous spinal epidural haematoma (SSEH) is a rare and serious condition. Four cases of SSEH are presented in this paper, three of which were in the cervical segment while one involved the entire spine. In two of these four cases the diagnosis was not made until late in the course, and persisting neurological deficits resulted; one case was diagnosed early and the neurological outcome was good; and in one the decompression achieved was inadequate, owing to intraoperative complications. Laminectomy was performed in all four cases for the purpose of decompression of the spinal cord but at different intervals after the onset of symptoms and with different neurological outcomes. The prognosis of SSEH depends heavily on the time that elapses between the onset of symptoms and the surgical treatment. The diagnostic procedure of choice is MRI. PMID- 17123046 TI - [Fracture of the distal ulna accompanying fracture of the distal radius. Minimally invasive treatment with elastic stable intramedullary nailing (ESIN)]. AB - The distal radius is one of the commonest sites of fracture, and this injury is sometimes associated with fracture of the distal ulna. In recent years, surgical treatment of distal radius fractures has consisted increasingly in internal fixation with locking plates followed by early functional postoperative treatment. The associated injury to the distal ulna has so far not received much attention in the literature. Various techniques have veen described for its treatment: Kirscher wire fixation, tension band wiring, and internal fixation with screws and plates. Following positive results with elastic stable intramedullary nailing (ESIN) in the treatment of shaft fractures in children this technique was also applied in in fractures in adults (forearm, clavicle). Use of this technique for stabilisation of distal ulnar fractures has not previously been reported. In the course of a prospective longitudinal study (EBM level II), in 26 patients with an average age of 73.6 (42-88 years), bone healing in anatomical position was achieved in all cases within 6-12 weeks after closed reduction and anterograde ESIN with subsequent treatment that did not involve immobilization. No length differences of more than 2 mm and no functionally relevant deviations of the ulnar axis were observed. Apart from 3 cases of nail perforation at the distal end of the ulna, which had no clinical manifestations, there were no complications. ESIN offers a minimally invasive option for the treatment of unstable fractures of the distal ulna associated with distal radius fractures; it allows functional aftertreatment and can be regarded at least as an alternative to open reduction with internal fixation. PMID- 17123047 TI - [Soft tissue carcinoma. Epidemiology, diagnostics and therapy]. AB - The treatment of soft tissue sarcoma requires an individually tailored, multimodal therapy due to the high variability in the clinical situation. Resection is the usual treatment for patients with superficial, low grade tumors with a diameter of <5 cm. For intermediate grade, differentiated lesions, resection with negative resection edges combined with radiotherapy attains an almost 80% total survival rate. For patients with high grade sarcoma of >5 cm, local control can be attained by resection and radiotherapy, however every second patient will develop metastases. Patients with a local recurrence should consider a new resection. Radiotherapy is the more effective the lower the remaining postoperative tumor burden. PMID- 17123048 TI - [High-dose 106Ruthenium plaque brachytherapy for posterior uveal melanoma. A clinico-pathologic study]. AB - BACKGROUND: The purpose of this study was to report the multifactorial results of high-dose (106)Ruthenium plaque brachytherapy for (cilio-)choroidal melanoma and to confirm them by histological examinations. PATIENTS AND METHODS: 100 patients with choroidal or ciliochoroidal melanoma treated by high-dose 106Ruthenium plaque brachytherapy were followed-up for 5 years. 12 secondary enucleated eyes were compared to a non-irradiated matched group by light microscopy. RESULTS: The 5-year local tumour control rate was 93%, the 5-year survival rate 91%. Late radiogenic side effects occured as a retinopathy in 13%, as an optic neuropathy in 5% and as a secondary glaucoma in 3% of the patients. 14% had to be enucleated, 10% developed metastases. The histopathologic examination revealed significantly higher degrees of necrosis (p=0,041), balloon cell degeneration (p=0,025) and fibrosis (p<0,001) in the irradiated melanomas than in the control tumours. CONCLUSION: High-dose 106Ruthenium plaque brachytherapy turned out to be an effective treatment procedure for posterior uveal melanoma (not exceeding a prominence of 5,5 mm) with a high rate of local tumour control and a low rate of side effects. PMID- 17123049 TI - [Obligatory informed consent on the risks to the accommodation ability of the eye after the implantation of an artificial lens]. AB - The article reports a medical malpractice claim in Germany against an ophthalmologist who implanted an artificial lens without informing the patient about the risk of losing the eye's accommodation ability before undertaking surgery. As informed consent was not obtained, in spite of the obligation to so, the court ruled that the physician should pay compensation for medical malpractice. The author shows how similar malpractice cases can be prevented in future. PMID- 17123050 TI - [Juxtapapillary capillary retinal angioma with epiretinal membrane of the macula in familial Von-Hippel-Lindau-Syndrome]. AB - A case of juxtapapillary capillary retinal angioma associated with a vascularized epiretinal membrane of the macula in a 6-year-old girl is presented. Von-Hippel Lindau-Syndrome was revealed by molecular genetic methods, and further family members were identified as gene carriers. The retinal angioma embedded in an epiretinal membrane was removed completely with the membrane by pars plana vitrectomy with a good functional result. Histopathology confirmed the diagnosis of capillary angioma. PMID- 17123051 TI - [Pseudoangiitis in bilateral ocular ischemia]. AB - BACKGROUND: The term 'frosted branch angiitis' was initially used to describe an idiopathic form of vasculitis in association with panuveitis. It has since also been used to describe the angiographic phenomenon of diffuse leakage along retinal vessels against the background of other ocular and systemic diseases. METHODS: We describe a 57-year-old male patient with acute bilateral reduction of visual acuity, headaches and absence of any pulse at the temporal arteries. Fluorescence angiography showed bilateral diffuse leakage along all the retinal vessels, which resembled frosted branches. Laboratory parameters and histology were not indicative of vasculitis. Imaging showed complete occlusion of both common carotid arteries and a hypoplastic vertebral artery on the left. CONCLUSIONS: Ocular ischemia may imitate primarily inflammatory conditions in its angiographic appearance. PMID- 17123052 TI - [Subtle, temporary, homonymous visual field defect--fatal causes?]. PMID- 17123053 TI - [Imaging methods in rheumatology. Current status of knowledge and future aspects]. PMID- 17123054 TI - [Therapy of masticatory muscle pain: recommendations for clinical management]. AB - Myalgias of the masticatory muscles are the most frequent noninfectious complaints in the orofacial region. After summarizing the current knowledge on the physiology, etiology, pathophysiology, diagnosis, and differential diagnosis of masticatory muscle pain, we specifically focus on management recommendations. Results of an extensive literature search show that for the majority of patients pain reduction or pain relief can be achieved with noninvasive reversible methods. Longitudinal short- and long-term studies have revealed that different therapeutic measures are similarly effective. In patients with chronic masticatory myalgias associated with psychosocial impairment, however, additional involvement of a psychotherapist is crucial. PMID- 17123055 TI - Symptomatic vertebral hemangioma related to pregnancy. AB - In this paper, we report a case of vertebral hemangioma during pregnancy in a 21 year-old woman presenting with paraparesis of rapid onset. An emergency MRI scan of the dorsal spine showed a lesion of the ninth thoracic vertebra with extradural extension and marked spinal cord compression. A cesarean section was done, and this was followed by emergent laminectomy. Her symptoms and neurologic deficits quickly improved. The etiopathogenesis, clinical, radiological features, and treatment modalities are discussed in the light of the literature. PMID- 17123058 TI - Appropriateness of out-of-hours CT head scans. AB - This study was conducted to evaluate the appropriateness of the out-of-hours computed tomography (CT) head scans (scans performed outside normal working hours) in a large district general hospital in the United Kingdom. CT scan request forms and CT reports of adult patients were reviewed who had their CT head scans done between 1700 and 0900 h the next day and all weekend and national holidays in the study period of 4 months. Information regarding change in the patient management resulting from CT scan was extracted from review of patients' case notes. An urgent CT head scan request was deemed appropriate if it led to an immediate change in a patient's management. Appropriateness of the requests according to the various guidelines was also evaluated. One hundred and twenty one patients were included in the study. Majority (70%) of the scans were requested by the emergency department. Eighty-one (66%) scans were both requested and performed between 1700 and 2200 h. Immediate change in management of 80% patients occurred. In 20 (17%), the change in management occurred only after 0900 h the next day and in 4 (3%), management did not change at all. Common scenarios involving change in patient's management included intracranial bleed requiring urgent neurosurgical intervention and hemorrhagic stroke being ruled out. Out-of hours CT head scans also facilitated early discharge of the patients with head injuries and headache from the hospital. This retrospective study has found that a high proportion of out-of-hours CT head scans were appropriate and led to change in the patient's immediate medical care. PMID- 17123059 TI - Some speculation on the origin of glioblastoma. AB - Glioblastoma, the most common primary brain tumor, is also the most deadly, with median survival of about one year, which is little improved over the last five decades. Its pathogenesis is a vexing problem. Despite extensive basic and clinical scientific research, little is known regarding the cause of this disease, the genetic factors which drive its course, or any strategies which may result in effective treatment. This persistent resistance to understanding suggests to the authors that some of the fundamental assumptions regarding the disease are likely to be flawed, and that a new paradigm must be sought to replace them. This manuscript is a review of some of what is known regarding this disease, and then presents a series of hypotheses which compromise an alternative view of glioblastoma. PMID- 17123060 TI - Inclusion-induced boundary layers in lipid vesicles. AB - The equilibrium shapes of lipid vesicles are perturbed by rigid inclusions. In a two-dimensional vesicle, that may also model a cylindrically elongated tubule, the shape modifications can be determined analytically, and turn out to be significant even far from the inclusion. On the contrary, previous numerical work has given evidence that in the three-dimensional case the shape perturbations decay quite rapidly and are negligible a few inclusion radii away. In this paper, we use the tools of asymptotic analysis to derive analytically the shape of the boundary layer induced by the inclusion. As a result, we are able to determine the dominant part of the free-energy perturbation that, in turn, allows to identify the vesicle points where the inclusion prefers to sit. PMID- 17123061 TI - On the definition and modeling of incremental, cumulative, and continuous growth laws in morphoelasticity. AB - In the theory of elastic growth, a growth process is modeled by a sequence of growth itself followed by an elastic relaxation ensuring integrity and compatibility of the body. The description of this process is local in time and only corresponds to an incremental step in the total growth process. As time evolves, these incremental growth steps are compounded and a natural question is the description of the overall cumulative growth and whether a continuous description of this process is possible. These ideas are discussed and further studied in the case of incompressible shells. PMID- 17123062 TI - High levels of linkage disequilibrium and associations with forage quality at a phenylalanine ammonia-lyase locus in European maize (Zea mays L.) inbreds. AB - Forage quality of maize is influenced by both the content and structure of lignin in the cell wall. Phenylalanine Ammonia-Lyase (PAL) catalyzes the first step in lignin biosynthesis in plants; the deamination of L-phenylalanine to cinnamic acid. Successive enzymatic steps lead to the formation of three monolignols, constituting the complex structure of lignin. We have cloned and sequenced a PAL genomic sequence from 32 maize inbred lines currently employed in forage maize breeding programs in Europe. Low nucleotide diversity and excessive linkage disequilibrium (LD) was identified at this PAL locus, possibly reflecting selective constrains resulting from PAL being the first enzyme in the monolignol, and other, pathways. While the association analysis was affected by extended LD and population structure, several individual polymorphisms were associated with neutral detergent fiber (not considering population structure) and a single polymorphism was associated with in vitro digestibility of organic matter (considering population structure). PMID- 17123063 TI - Evolution of chloroplast mononucleotide microsatellites in Arabidopsis thaliana. AB - The level of variation and the mutation rate were investigated in an empirical study of 244 chloroplast microsatellites in 15 accessions of Arabidopsis thaliana. In contrast to SNP variation, microsatellite variation in the chloroplast was found to be common, although less common than microsatellite variation in the nucleus. No microsatellite variation was found in coding regions of the chloroplast. To evaluate different models of microsatellite evolution as possible explanations for the observed pattern of variation, the length distribution of microsatellites in the published DNA sequence of the A. thaliana chloroplast was subsequently used. By combining information from these two analyses we found that the mode of evolution of the chloroplast mononucleotide microsatellites was best described by a linear relation between repeat length and mutation rate, when the repeat lengths exceeded about 7 bp. This model can readily predict the variation observed in non-coding chloroplast DNA. It was found that the number of uninterrupted repeat units had a large impact on the level of chloroplast microsatellite variation. No other factors investigated- such as the position of a locus within the chromosome, or imperfect repeats- appeared to affect the variability of chloroplast microsatellites. By fitting the slippage models to the Genbank sequence of chromosome 1, we show that the difference between microsatellite variation in the nucleus and the chloroplast is largely due to differences in slippage rate. PMID- 17123065 TI - FR180204, a novel and selective inhibitor of extracellular signal-regulated kinase, ameliorates collagen-induced arthritis in mice. AB - Extracellular signal-regulated kinase (ERK), a serine/threonine protein kinase of the mitogen-activated protein kinase superfamily, is activated by various stimuli in inflammatory cells. We recently described FR180204 (5-(2-phenylpyrazolo[1,5 a]pyridin-3-yl)-1H-pyrazolo[3,4-c]pyridazin-3-amine), a novel selective ERK inhibitor. In this paper, we investigated the effect of FR180204 on collagen induced arthritis (CIA) in DBA/1 mice, an animal model of rheumatoid arthritis (RA) mediated by type II collagen (CII)-reactive T cells and anti-CII antibodies. Preventive administration of FR180204 (100 mg/kg, i.p., b.i.d.) significantly ameliorated the clinical arthritis and body weight loss occurring in the CIA mice. Further, FR180204-treated mice showed a significant decrease in plasma anti CII antibody levels (62%). FR180204 also attenuated delayed-type hypersensitivity in CII-immunized DBA/1 mice, an inflammatory response elicited by CII-reactive T cells, in a dose-dependent manner (52 and 62% inhibition at 32 and 100 mg/kg, respectively). Moreover, FR180204 inhibited in vitro CII-induced proliferation of lymph node cells prepared from CII-immunized mice, in which CII-specific T cells are known to undergo specific proliferation. In conclusion, our results suggest that ERK regulates both the cell-mediated and humoral immune responses in the development of CIA. ERK inhibitors may be useful as therapeutic reagents for the treatment of RA. PMID- 17123066 TI - Probing traces of hydrogen peroxide by use of a biosensor based on mediator-free DNA and horseradish peroxidase immobilized on silver nanoparticles. AB - A new electrochemical biosensor for determination of hydrogen peroxide (H(2)O(2)) has been developed by immobilizing horseradish peroxidase (HRP) on silver colloids (nanosilver) and use of a DNA-functionalized interface. In the presence of the DNA and the nanosilver the immobilized HRP gives a pair of well-defined redox peaks with an electron-transfer rate constant of 3.27 +/- 0.91 s(-1) in pH 7.0 PBS. The presence of DNA also provides a biocompatible microenvironment for enzyme molecules, greatly amplifies the amount of HRP molecules immobilized on the electrode surface, and improves the sensitivity of the biosensor. Under optimum conditions the biosensor has electrocatalytic activity in the reduction of hydrogen peroxide with linear dependence on H(2)O(2) concentration in the range 1.5 x 10(-6) to 2.0 x 10(-3) mol L(-1); the detection limit is 5.0 x 10(-7) mol L(-1) at a signal-to-noise ratio of 3. The K(app)(m) value of HRP in the composite membrane was found to be 1.62 mmol L(-1). These results suggest that the properties of the complex film, with its bioelectrochemical catalytic activity, could make it useful for development of bioelectronic devices and for investigation of protein electrochemistry at functional interfaces. PMID- 17123067 TI - Spectroscopic study of the interaction of actinomycin D with oligonucleotides carrying the central base sequences -XGCY- and -XGGCCY- using multivariate methods. AB - The interactions of actinomycin D (ACTD) with the oligonucleotides 5'-CAAAGCTTTG 3', 5'-CATGGCCATG-3' and 5'-TATGGCCATA-3' were investigated by means of acid-base titrations and mole-ratio and melting experiments monitored by molecular absorption and circular dichroism (CD) spectroscopies. For each experiment, CD and molecular absorption spectra were recorded at each point in the experiment, and later analyzed via appropriate multivariate data analysis methods. The study of the interactions between these oligonucleotides and ACTD at 25 degrees C showed the formation of an interaction complex with a stoichiometry of 1:1 (ACTD:duplex) and values for the log(formation constant) of 5.1+/-0.3, 6.4+/-0.2, and 5.6+/-0.2, respectively. An additional interaction complex at higher temperatures was also detected, which might be related to the single-stranded forms of the oligonucleotides. PMID- 17123068 TI - The use of Raman spectroscopy to provide an estimation of the gross biochemistry associated with urological pathologies. AB - Near-infrared Raman spectroscopy, an optical technique that is able to interrogate biological tissues, has been used to study bladder and prostate tissues, with the objective being to provide a first approximation of gross biochemical changes associated with the process of carcinogenesis. Prostate samples for this study were obtained by taking a chip at TURP, and bladder samples from a biopsy taken at TURBT and TURP, following ethical approval. Spectra were taken from purchased biochemical constituents and different pathologies within the bladder and the prostate. We were then able to determine the biochemical basis for these pathologies by utilising an ordinary least squares fit. We have shown for the first time that we are able to utilise Raman spectroscopy in determining the biochemical basis for the different pathologies within the bladder and prostate gland. In this way we can achieve a better understanding of disease processes such as carcinogenesis. This could have major implications in the future of the diagnosis of disease within the bladder and the prostate gland. PMID- 17123069 TI - Updated estimates of the selenomethionine content of NIST wheat reference materials by GC-IDMS. AB - Updated estimates of the selenomethionine content of four NIST wheat reference materials have been obtained by use of a revised gas chromatography-stable isotope dilution mass spectrometric method. The revised method makes use of digestion with methanesulfonic acid, which enables more complete recovery of endogenous selenomethionine than was previously achieved by overnight denaturing treatment in 0.1 mol L(-1) HCl. The NIST wheat reference materials each contain approximately 55% of their total Se content as selenomethionine. Information about forms of Se in reference materials adds value to these materials in Se speciation studies. Estimates of selenomethionine content are also provided for other wheat samples, including several grown under conditions of exposure to high Se levels. These samples also contain approximately 55% of their total Se content as selenomethionine. The consistent level of 55% of total selenium occurring in the form of selenomethionine when the total selenium content varies by a factor of 500 is suggestive of an active mechanism of incorporation of selenium into wheat grain. Figure Selenomethionine content of wheat samples. PMID- 17123070 TI - Cortical activation to indoor versus outdoor scenes: an fMRI study. AB - Prior studies identify two cortical areas, posterior parahippocampal cortex and retrosplenial cortex, that preferentially activate to images of real-world scenes compared to images of other meaningful visual stimuli such as objects and faces. Behavioral and computational studies suggest that sub-categories of real-world scenes differ in their visual and semantic properties. It is presently unknown whether the cortical areas that have been implicated in scene analysis similarly activate differentially to behaviorally relevant scene sub-categories. To examine this issue, we directly compared cortical activation to indoor and outdoor scenes in an fMRI study with a large number of non-repeated images in each condition. Activation in posterior parahippocampal cortex, including parahippocampal place area, was significantly greater for indoor than outdoor scenes. In contrast, no such difference was observed in retrosplenial cortex, though this region preferentially activated to scenes over faces. These findings suggest differences in function in these two areas. The results are consistent with the view that posterior parahippocampal cortex is functional in processing local space. PMID- 17123071 TI - Preclinical testing of a new clot-retrieving wire device using polyvinyl alcohol hydrogel vascular models. AB - INTRODUCTION: Cerebral embolism is the principal cause of cerebral infarction. Recently, mechanical embolectomy has been proposed as an effective method. We performed a preclinical evaluation of a new mechanical clot-retrieving wire. METHODS: This clot-retrieving wire consisted of three nitinol loops at the tip of a microguidewire. These three loops could be collapsed into a 0.018-inch wire compatible microcatheter. Each loop was 8 mm long and 3.5 mm wide. For simulation, polyvinyl alcohol (PVA) vascular anatomical models of the human carotid (eight models) and vertebrobasilar (three models) circulation were constructed. A pulsatile flow circulation system was used. Embolic clots were produced using pig blood plasma. The microcatheter and the microguidewire were advanced beyond the clot. The wire was then exchanged for the retrieving wire. The microcatheter was then pulled slightly back to open the loops. The clot was then caught by withdrawal of the system. Once caught, the clot was retrieved to the guiding catheter tip. We investigated the following points: ease of device deployment, clot capture ability, clot removal against blood flow and removal of the clot out of the introducer system. RESULTS: A total of 104 procedures were performed in 11 PVA models and evaluated. The drop rate was 19%. We succeeded in partial and total recanalization in 51.0% of the procedures (53/104) within 30 minutes. CONCLUSION: This new clot-retrieving wire could be useful for mechanical clot extraction in stroke. PMID- 17123072 TI - Transvenous embolization of a dural carotid-cavernous fistula through the contralateral superior petrosal sinus. AB - We report a patient with progressive ophthalmological problems related to a dural carotid-cavernous fistula who was successfully treated endovascularly. Cavernous sinus catheterization was achieved through the contralateral superior petrosal sinus, permitting occlusion of the fistula without complications. PMID- 17123073 TI - The 53-kDa proteolytic product of precursor starch-hydrolyzing enzyme of Aspergillus niger has Taka-amylase-like activity. AB - The 53-kDa amylase secreted by Aspergillus niger due to proteolytic processing of the precursor starch-hydrolyzing enzyme was resistant to acarbose, a potent alpha glucosidase inhibitor. The enzyme production was induced when A. niger was grown in starch medium containing the inhibitor. Antibodies against the precursor enzyme cross-reacted with the 54-kDa Taka-amylase protein of A. oryzae. It resembled Taka-amylase in most of its properties and also hydrolyzed starch to maltose of alpha-anomeric configuration. However, it did not degrade maltotriose formed during the reaction and was not inhibited by zinc ions. PMID- 17123074 TI - Expression, refolding, and characterization of recombinant thrombopoietin/stem cell factor fusion protein in Escherichia coli. AB - Thrombopoietin/stem cell factor (TPO/SCF) is a novel fusion protein that combines the complementary biological effects of TPO and SCF into a single molecule. In this study, TPO/SCF gene was cloned into pET32a and expressed as a thioredoxin (Trx) fusion protein with a C-terminal 6His-tag in Escherichia coli BL21(DE3) under the control of T7 promoter. Trx-TPO/SCF protein approximately accounted for 20% of the total bacterial proteins and was found to accumulate in inclusion bodies. Inclusion bodies were separated from cellular debris, washed with buffer containing 2 M urea, and solubilized with 8 M urea. The refolding of Trx-TPO/SCF was then carried out by an on-column method. Soluble Trx-TPO/SCF was characterized for its dose-dependent effects on promoting cells proliferation in both TF1 and Mo7e cell lines. rhTPO/SCF was released by thrombin digestion and further purified by Ni(2+) affinity chromatography. Western blot analysis confirmed the identities of Trx-TPO/SCF and rhTPO/SCF. PMID- 17123075 TI - The use of Tetrahymena thermophila mutant cell line for removal of cholesterol from milk. AB - The nonpathogenic ciliate Tetrahymena thermophila converts cholesterol from foodstuffs into provitamin D compounds in high yields. However, prolonged incubation with wild-type strain CU-399 at high densities results in a final deterioration of milk properties, possibly as a result of secreted hydrolases. Here we attempted to solve this problem using MS-1 Tetrahymena strain, a stable mutant with a low rate of hydrolase secretion. Densities of to 2 x 10( 6 ) cells/ml can be incubated for up to 5 h in milk, without any clotting or change in appearance. Moreover, centrifugation of this suspension eliminates most of the cells, and results in an about 75% +/- 10 (n = 10) decrease of the initial cholesterol. Sterols are recovered in the cell pellets, which show that Tetrahymena is able to avidly capture them from the medium. Therefore, this mutant strain is optimal for milk cholesterol depletion, avoiding unfavorable sensory alterations. PMID- 17123076 TI - Utilization of hydrophobic bacterium Rhodococcus opacus B-4 as whole-cell catalyst in anhydrous organic solvents. AB - Rhodococcus opacus strain B-4, which has recently been isolated as an organic solvent-tolerant bacterium, has a high hydrophobicity and exhibits a high affinity for hydrocarbons. This bacterium was able to survive for at least 5 days in organic solvents, including n-tetradecane, oleyl alcohol, and bis(2 ethylhexyl) phthalate (BEHP), which contained water less than 1% (w/v). The biocatalytic ability of R. opacus B-4 was demonstrated in the essentially nonaqueous BEHP using indigo production from indole as a model conversion. By the catabolism of oleic acid for NADH regeneration, indigo production increased up to 71.6 microg ml(-1) by 24 h. PMID- 17123077 TI - The inhibition of aggregation of recombinant human consensus interferon-alpha mutant during Pichia pastoris fermentation. AB - Lower induction temperature and polyoxyethylene sorbitan monolaurate (Tween-20) were successfully used to inhibit the aggregation of recombinant human consensus interferon-alpha mutant (cIFN) during Pichia pastoris fermentation. When the induction temperature was decreased from 30 to 20 degrees C, the cIFN secreted into the medium was in the form of monomers instead of aggregates. The maximum specific activity at 20 degrees C was 4.04 times as high as that at 30 degrees C. There was no obvious effect on the cell growth at 20 degrees C, but the total protein level was decreased. Similar inhibition effect on cIFN aggregation was observed when 0.2 g l(-1) Tween-20 was added during induction. Furthermore, there was a synergistic effect found between induction temperature and Tween-20 on the inhibition of cIFN aggregation. The maximum specific activity with Tween-20 at 20 degrees C was 19.9-fold higher than that without Tween-20 at 30 degrees C. PMID- 17123078 TI - Use of protein chip mass spectrometry to monitor biotinylation reactions. AB - Surface-enhanced laser desorption/ionization time-of-flight analysis was used to monitor both the kinetics and heterogeneity of product formation during the biotinylation of a number of model proteins and peptide targets. The selected molecules were the IgG-binding protein, protein A, human serum albumin, and a synthetic peptide corresponding to the N terminus of a streptococcal M1 protein. The extent of biotinylation was determined by kinetic analysis of the shift in molecular mass from the native material. Each residue modified by reaction with N hydroxysuccinimide biotin resulted in an addition of approximately 341 amu to the native protein or polypeptide. The novelty of the method was in the ability to determine the molecular mass shift, without first separating the targeted molecule from the biotinylating reagent. The analysis was rapid, simple, and provided information on the average number of biotin molecules added and the homogeneity of the resulting product. PMID- 17123079 TI - PHA synthase engineering toward superbiocatalysts for custom-made biopolymers. AB - Poly-3-hydroxyalkanoates [P(3HA)s] are biologically produced polyesters that have attracted much attention as biodegradable polymers that can be produced from biorenewable resources. These polymers have many attractive properties for use as bulk commodity plastics, fishing lines, and medical uses that are dependent on the repeating unit structures. Despite the readily apparent benefits of using P(3HA)s as replacements for petrochemical-derived plastics, the use and distribution of P(3HA)s have been limited by their cost of production. This problem is currently being addressed by the engineering of enzymes involved in the production of P(3HA)s. Polyhydroxyalkanoate (PHA) synthase (PhaC) enzymes, which catalyze the polymerization of 3-hydroxyacyl-CoA monomers to P(3HA)s, were subjected to various forms of protein engineering to improve the enzyme activity or substrate specificity. This review covers the recent history of PHA synthase engineering and also summarizes studies that have utilized engineered PHA synthases. PMID- 17123080 TI - Identification of advanced coronary artery disease with exercise myocardial perfusion imaging: the clinical value of a novel approach for assessing lung thallium-201 uptake. AB - INTRODUCTION: The precise clinical utility of lung 201Tl uptake in exercise SPECT myocardial perfusion imaging remains open to research. This study validates an optimal index for lung 201Tl uptake measurement and assesses its value in the prediction of higher-risk coronary artery disease (CAD). METHOD: Three hundred and ninety-eight patients underwent exercise SPECT myocardial perfusion imaging. They were separated into derivation (n = 217) and validation (n = 186) groups, both including sub-populations of lower- and higher-risk CAD, according to coronary angiography. Another 56 individuals with a low probability of CAD comprised the control group. From a planar, anterior, post-exercise acquisition, the lung (L) to heart (H) maximal (L/H(max)), total (L/H(mean)) and background subtracted total (L/H(net)) ratios were calculated. These were also adjusted for confounding variables, as assessed from the control group. ROC analysis was used to compare the L/H ratios and define thresholds of abnormality. The performance of the optimal index was assessed in the derivation group and was then tested in the validation population. Subsequently, it was compared with other scintigraphic, exercise electrocardiography and clinical variables. RESULTS: In the derivation group L/H(net) was a better discriminator for higher-risk CAD than both L/H(max) and L/H(mean). Similarly, the adjusted L/H(net) was a better discriminator than both the adjusted L/H(max) and the adjusted L/H(mean). No significant difference was attained between L/H(net) and the adjusted L/H(net). At the upper defined threshold of abnormality, sensitivity and specificity of L/H(net) in the detection of higher-risk CAD in the derivation and the validation cohorts were 52% and 92% versus 47% and 94%, respectively (p = ns). The results were similar at other defined thresholds. Moreover, L/H(net) was found to be a significant predictor of higher-risk CAD, superior to myocardial perfusion images, transient ventricular dilation measurements, and clinical and exercise testing variables (ROC analysis and logistic regression). By raising the threshold of abnormality of L/H(net), specificity and positive predictive value increased, whereas sensitivity and negative predictive value decreased. CONCLUSION: Lung 201Tl assessment assists substantially in the identification of higher-risk CAD in exercise SPECT myocardial perfusion imaging and this is best achieved by L/H(net). This index is a significant predictor of higher-risk CAD, superior to myocardial perfusion images, and its value is associated with the probability of a disease state. PMID- 17123081 TI - Effects of rhBMP-2 on cortical strut allograft healing to the femur in revision total hip arthroplasties: an experimental study. AB - We have studied the effects of recombinant human bone morphogenetic protein-2 (rhBMP-2) on cortical strut allograft healing and remodelling in revision hip arthroplasty. Thirty adult New Zealand rabbits underwent bilateral onlay allograft strut procedures to the femur using wires. The left femur (experimental side) received the rhBMP-2 device (1.0-mg rhBMP-2/gelatin composites) interposed between the allograft and host bone, while the right side was grafted with an allograft strut as the control. The femurs and implants were retrieved at 4, 6, and 8 weeks postoperatively. The healing of cortical strut grafts to the femur was enhanced dramatically by the addition of the rhBMP-2 device in radiographic examination, contact radiographic examination, non-decalcification sections, fluorescence tag, and computer-aided image analysis. The remodelling of cortical strut allograft was also accelerated. The new bone formation ratio and radiographic scores of the experimental side were also much higher than the control side at all times. Strut healing with the rhBMP-2 device at 4 weeks postoperatively was superior to the healing in control sides at 8 weeks. Our findings showed that the rhBMP-2 device improved and accelerated the course of cortical strut allograft healing and remodelling with host bone. PMID- 17123082 TI - Pure red cell aplasia in a patient of chronic granulocyte leukemia associated with imatinib. PMID- 17123083 TI - Lack of clinical efficacy of rituximab in the treatment of autoimmune neutropenia and pure red cell aplasia: implications for their pathophysiology. AB - We describe 11 patients with severe refractory autoimmune cytopenias treated with the anti-CD20 monoclonal antibody rituximab. Six patients had autoimmune neutropenia (AIN), two had pure red cell aplasia (PRCA), one had AIN and autoimmune haemolytic anaemia, one had AIN and immune thrombocytopaenia purpura (ITP) and one had PRCA and ITP. Rituximab was administered at a dose of 375 mg/m(2) as an intravenous infusion weekly for 4 weeks. Six of eight patients with AIN and all three patients with PRCA did not respond. Two patients died: one with resistant AIN and autoimmune haemolytic anaemia died of pneumocytis pneumonia infection, and one with PRCA and ITP died of an acute exacerbation of bronchiectasis. Rituximab in AIN and PRCA appears to be less effective than Campath-1H when compared to historical data from our group. This supports the hypothesis that T cells may be important in the pathophysiology of AIN and PRCA. PMID- 17123084 TI - Asymptotic stability of equilibria of selection-mutation equations. AB - We study local stability of equilibria of selection-mutation equations when mutations are either very small in size or occur with very low probability. The main mathematical tools are the linearized stability principle and the fact that, when the environment (the nonlinearity) is finite dimensional, the linearized operator at the steady state turns out to be a degenerate perturbation of a known operator with spectral bound equal to 0. An example is considered where the results on stability are applied. PMID- 17123085 TI - Stochastic stable population growth in integral projection models: theory and application. AB - Stochastic matrix projection models are widely used to model age- or stage structured populations with vital rates that fluctuate randomly over time. Practical applications of these models rest on qualitative properties such as the existence of a long term population growth rate, asymptotic log-normality of total population size, and weak ergodicity of population structure. We show here that these properties are shared by a general stochastic integral projection model, by using results in (Eveson in D. Phil. Thesis, University of Sussex, 1991, Eveson in Proc. Lond. Math. Soc. 70, 411-440, 1993) to extend the approach in (Lange and Holmes in J. Appl. Prob. 18, 325-344, 1981). Integral projection models allow individuals to be cross-classified by multiple attributes, either discrete or continuous, and allow the classification to change during the life cycle. These features are present in plant populations with size and age as important predictors of individual fate, populations with a persistent bank of dormant seeds or eggs, and animal species with complex life cycles. We also present a case-study based on a 6-year field study of the Illyrian thistle, Onopordum illyricum, to demonstrate how easily a stochastic integral model can be parameterized from field data and then applied using familiar matrix software and methods. Thistle demography is affected by multiple traits (size, age and a latent "quality" variable), which would be difficult to accommodate in a classical matrix model. We use the model to explore the evolution of size- and age-dependent flowering using an evolutionarily stable strategy (ESS) approach. We find close agreement between the observed flowering behavior and the predicted ESS from the stochastic model, whereas the ESS predicted from a deterministic version of the model is very different from observed flowering behavior. These results strongly suggest that the flowering strategy in O. illyricum is an adaptation to random between-year variation in vital rates. PMID- 17123086 TI - Descending control of turning behavior in the cockroach, Blaberus discoidalis. AB - Legged locomotion has evolved as the most effective form of movement through unpredictable and tortuous environments. Upon encountering an obstacle, an animal must evaluate the object with its sense organs then use the information it acquires to direct appropriate transitional behaviors, such as turning. Previous studies using genetic and surgical lesions implicate the central body complex (CBC) in control of such transitional behaviors of various insects. In this study, lesions of the CBC and surrounding brain regions were used to examine the effects of damage on turning in free-moving and tethered cockroaches. Lesions were performed either as sagittal incisions or by inserting small pieces of foil into regions of the brain. Locomotor behaviors of intact and lesioned animals were compared using high speed video and kinematic analysis. The lesions locations were determined through histological methods. Sagittal lesions to the CBC often result in continuous or incorrect turns. Foil lesions in the CBC also increase the probability that individuals will show turning deficits. The location and degree of the lesion had a strong effect on the animal's ability to turn. These data strongly suggest that the CBC mediates the effects of head sense organs that produce changes in the direction of walking. PMID- 17123087 TI - The parasitic mite Varroa destructor affects non-associative learning in honey bee foragers, Apis mellifera L. AB - The parasitic mite Varroa destructor influences flight behavior, orientation and returning success of forager honeybees (Apis mellifera) infested as adults. As impaired orientation toward the nest entrance might be due to deficiency in recognition and responsiveness to stimuli in the environment, we examined effects of V. destructor on sensory responsiveness, non-associative and associative learning of honey bee foragers by using proboscis extension reaction paradigm (PER). Although infested and uninfested workers were initially equally responsive to different concentrations of sugar water, we found differences in non associative learning. In habituation, PER to repeated sugar stimulation of the antennae occurred faster in infested foragers compared to uninfested foragers. In sensitization, infested foragers showed a lower response to an odor stimulus following sugar stimulation than non-infested foragers. Differences in non associative paradigms were more pronounced in bees with lower responsiveness to sucrose. In conditioning learning experiments, a significant reduction in proboscis extension response was found 1 min but not 12 min after a single conditioning trial indicating that V. destructor predominantly affects the non associative components of learning and its underlying neural and molecular processes. PMID- 17123088 TI - Postactivation inhibition of spontaneously active neurosecretory neurons in the medicinal leech. AB - Spontaneously active neurosecretory neurons in vertebrate and invertebrate nervous systems share similarities in firing frequencies, spike shapes, inhibition by the transmitters they themselves release and postactivation inhibition, an intensity-dependent period of suppressed spontaneous generation of action potentials following phases of high-frequency activity. High-frequency activation of spontaneously active serotonin-containing Retzius cells in isolated ganglia of the leech Hirudo medicinalis induced prolonged membrane hyperpolarisations causing periods of postactivation inhibition of up to 33 s. The duration of the inhibitory periods was directly related to both the number and rate of spikes during activation and was inversely proportional to a cell's spontaneous firing frequency. The periods of postactivation inhibition remained unaffected by both serotonin depletion through repeated injections of 5,7 dihydroxytryptamine and suppressing the afterhyperpolarisation following each action potential with tetraethylammonium (TEA), iberiotoxin or charybdotoxin, suggesting that neither autoinhibition by synaptic release of serotonin nor calcium-activated potassium channels contribute to the underlying mechanism. In contrast, the postactivation inhibitory period was significantly affected both by differential electrical stimulation of the same Retzius cells via microelectrodes filled with molar concentrations of either Na(+)-acetate or K(+)-acetate, and by partial inhibition of Na(+)/K(+)-ATPase with ouabain. Thus, postactivation inhibition in Retzius cells results from prolonged hyperpolarising activity of Na(+)/K(+)-ATPase stimulated by the accumulation of cytosolic Na(+ )during phases of high-frequency spike activity. PMID- 17123089 TI - Response of plasma and gastrointestinal melatonin, plasma cortisol and activity rhythms of European sea bass (Dicentrarchus labrax) to dietary supplementation with tryptophan and melatonin. AB - Melatonin is an effective antioxidant, immunostimulant, gonadal maturating regulator and antistress indoleamine that may be potentially useful for fish farmers. We have explored two possible ways of increasing plasma melatonin levels through the diet: direct melatonin supplementation (ME diet) and supplementation with the melatonin precursor tryptophan (TRP diet). To this end, a group of sea bass was fed a commercial diet (STD diet) at a regular time for 16 days, after which plasma, intestine, and bile samples were taken at four different time points: 120 min before, and 15, 180 and 480 min after feeding. Locomotor activity, intestinal and biliary melatonin, and plasma melatonin, serotonin and cortisol levels were measured. This same sampling process and analyses were also carried out after feeding sea bass TRP diet or ME diet for 1 week. Our results show that melatonin, but not tryptophan supplementation of the diet increases plasma, intestine and bile levels of melatonin. Plasma serotonin levels, on the other hand, were increased by dietary tryptophan, but not by melatonin, confirming the availability of supplemented tryptophan for serotonin synthesis. Both treatments were equally effective in reducing the high cortisol levels observed with the STD diet. PMID- 17123090 TI - No prognostic impact of isolated lymphovascular invasion after radical resection of rectal cancer--results of a multicenter observational study. AB - BACKGROUND: The prognostic impact of isolated lymphovascular invasion (LVI) after radical resection of rectal cancer is controversially discussed. However, it could be relevant to decide for an adjuvant treatment. AIM: The aim of the analysis was, based on the data of an observational study, to determine the prognostic relevance of the isolated LVI. MATERIALS AND METHODS: Patients after radical resection of rectal cancer with no hemangioinvasion were subdivided in three groups: I-no LVI, no lymph node metastases (LNM); II-positive LVI, no LNM; III-positive LNM. Five-year local recurrence rate, distant metastases-free and disease-free survival were determined uni- and multivariate. RESULTS: Patients, n = 846, were studied (I, n = 471; II, n = 75; III, n = 300). The univariate comparison between the groups revealed the following 5-year results: local recurrence rate: 9.4 vs 10.0 vs 14.0%; distant metastases-free survival: 84.1 vs 82.5 vs 49.3%; disease-free survival: 83.2 vs 80.7 vs 45.5%. The differences between groups I and III were significant, but not between groups I and II. The determined higher disease-free survival rate in group II vs group III was significant (P = 0.041), but the differences in local recurrence rate and rate of distant metastases did not reach statistical significance. The multivariate analysis revealed no impact of the isolated LVI on the oncological outcome. CONCLUSION: The isolated LVI has no independent prognostic impact on the local recurrence rate and long-term survival after radical resection of rectal cancer. Based on this finding, no indication for an adjuvant treatment in these patients can be derived. PMID- 17123091 TI - Recurrent cytogenetic aberrations in central neurocytomas and their biological relevance. AB - Central neurocytomas are rare central nervous system neoplasms. Since the first description, approximately 500 cases of these tumors have been published to date. Nevertheless, only a limited number of genetic studies on these tumors have been reported. Here we investigated 20 "typical" central neurocytomas using array based comparative genomic hybridization (array-CGH) with the GenoSensor Array 300. The functional significance of detected chromosomal aberrations harboring potent candidate genes was also examined at the mRNA expression level. Each tumor examined displayed DNA copy-number aberrations (CNAs), and mean number of CNAs per tumor was 38.1 +/- 7.1 (range 19-53). Frequent gains were mapped at 2p24.1 22.1, 10q23.3-26.3, 11q23-25, and 18q21.3-qter. Frequent losses were identified at 1pter-36.3, 1p34.3, 6q13-21, 12q23-qter, 17p13.3, 17q11-23, and 20pter-12.3. There were 10 gained and 23 lost single DNA clones affecting >or=40% of samples tested. mRNA expression levels of 24 selected candidate genes harbored in these imbalanced clones were analyzed. MYCN, PTEN, and OR5BF1 were strongly overexpressed, whereas BIN1, SNRPN, and HRAS were found to be strongly underrepresented at the transcriptional level. Thus these data support that MYCN oncogene gain/overexpression accompanied by reduced expression of BIN1 tumor suppressor may contribute to central neurocytoma tumorigenesis. PMID- 17123092 TI - The brain-specific protein TPPP/p25 in pathological protein deposits of neurodegenerative diseases. AB - Immunohistochemical detection of protein components of pathological inclusions is widely used for neuropathological diagnosis of neurodegenerative disorders. However, different antibodies and antigen unmasking methods may account for variability between research studies and thus may affect diagnostic accuracy. Using two different antibodies raised against either a segment (184-200 aa) or the full length of human recombinant brain-specific tubulin polymerization promoting protein TPPP/p25, we immunohistochemically screened neurodegenerative disorders, both with and without pathological alpha-synuclein structures. We tested three different epitope unmasking methods, we applied laser confocal microscopy to evaluate double immunolabelling, and we compared the amount of structures exhibiting TPPP/p25 and alpha-synuclein immunoreactivity. We demonstrate that there are a variety of staining patterns depending on the epitope retrieval method and antibody used. The antibody raised against aa 184 200 segment of TPPP/p25 is better in immunolabelling the majority of alpha synuclein immunopositive neuronal and glial pathological profiles detectable in Parkinson's disease, diffuse Lewy-body disease, and multiple system atrophy, in addition to immunostaining some extracellular huntingtin immunoreactive structures, lipofuscin, and neuromelanin particles. In contrast, the one raised against the full-length human recombinant TPPP/p25 is more suitable to immunodetect normal oligodendrocytes. Exposition of the segment aa 184-200 of TPPP/p25 in the aggregates of pathological inclusions renders this antibody a reliable marker of all types of alpha-synucleinopathies and suggests a role for TPPP/p25 in the aggregation process of some neurodegenerative conditions. PMID- 17123093 TI - Antegrade intramedullary splinting or percutaneous retrograde crossed pinning for displaced neck fractures of the fifth metacarpal? AB - INTRODUCTION: It is generally accepted that severely displaced or malrotated neck fractures of the fifth metacarpal should be treated by closed reduction and pinning. As antegrade intramedullary splinting avoids adhesions of the extensor hood and provides intramedullary stability this technique seems to be advantageous in comparison with traditional retrograde percutaneous crossed pinning. We raised the question whether the antegrade technique would give clinical and radiological results superior to the retrograde technique and investigated this within a comparative clinical study. MATERIALS AND METHODS: Thirty patients with displaced neck fractures of the fifth metacarpal who received operative treatment were included in a retrospective cohort study. Data was analysed from medical records and a clinical and radiological outcome evaluation was performed. Fifteen patients had antegrade intramedullary splinting and fifteen patients retrograde percutaneous pinning. Median time for follow-up was 17 and 18 months, respectively. RESULTS: A significantly better outcome was found after antegrade splinting for ROM restriction of the metacarpophalangeal joint (0 degrees vs. -15 degrees ; P = 0.016), shortening (P = 0.029), pain (0 vs. 5; VAS 0-100; P = 0.026), and overall assessment by the Steel score (400 vs. 357; P = 0.001). There was no statistical difference in grip strength and functional assessment by the DASH score. CONCLUSION: From our clinical and radiological data we conclude that antegrade intramedullary splinting is superior to retrograde percutaneous crossed pinning and thus should be preferentially considered for displaced neck fractures of the fifth metacarpal. PMID- 17123094 TI - Vaginal carcinoma in a completely prolapsed uterus. A case report. AB - BACKGROUND: The development of genuine vaginal carcinoma onto a completely prolapsed uterus is a very rare condition to deal with. CASE: We report here the clinical characteristics of a patient with vaginal carcinoma associated with a third-degree prolapsed uterus. The 80-year-old patient was admitted with a completely prolapsed uterus. The cervix was clinically normal but on the nearby prolapsed vaginal wall a large exophytical hard lesion had been developed. Biopsy of the lesion revealed squamous carcinoma. TREATMENT: The treatment performed was radical vaginal hysterectomy and excision of the upper two-thirds of the vagina without pelvic lymphadenectomy, followed by external beam irradiation. The patient is alive, with no signs of the disease 3.5 years after surgery. CONCLUSION: Surgical and radiotherapeutic treatments can be effectively combined in patients with vaginal carcinoma and complete genital prolapse, in order to improve survival benefits and reduce morbidity. PMID- 17123095 TI - Development of tracheal prostheses made of porous titanium: a study on sheep. AB - Authors report the development of a biomaterial to be used for tracheal and laryngeal reconstruction. This experimentation follows the replacement of trachea in rats with porous titanium implants. The aim of the study is to test this type of prosthesis on sheep, whose trachea is of comparable size to that of humans. Six ewes were implanted with porous titanium implants after resection of 5 cm of trachea. The planned period for the implantation was from 3 to 6 months before the sacrifice of the animals for histological analysis. After a simple immediate postoperative course, the implantations developed complications of tracheal patency, responsible for four deaths (tracheal obstruction by mucous plug n = 2, inferior necrosis of trachea n = 1, pneumopathy n = 1). The two remaining sheep presented no complications. The mechanical performance of the prostheses was good. The histological results showed an inflammatory stenosis of the tracheo prosthetic junctions, which was not the direct result of death. The protheses were integrated by the surrounding tissue, but endoprosthetic colonisation by pseudostratified ciliated columnar epithelium was low or nil. The absence of endoprosthetic lining was responsible for the complications. The biocompatibility of the biomaterial is not in question, but the surgical procedure will have to be modified by an endoprosthetic mucous graft before implantation so as to accelerate healing process. PMID- 17123096 TI - Extrusion forces of resorbable tacks and titanium screws in laryngeal chondrosynthesis. AB - Laryngotracheal trauma, partial laryngectomy and phonosurgery may necessitate reconstruction of the cartilaginous skeleton to ensure the quality of respiration and voice. The present report focuses on initial experience gained with a new resorbable material for plates and tacks that allows chondrosynthesis of the laryngeal skeleton. A comparison of the extrusion forces necessary to pull out the resorbable tacks versus conventional titanium screws and the degree of deformation until failure represent the experimental parameters of reconstruction quality under investigation. The PolyMax system (Synthes, Oberdorf, Switzerland) was used in a human cadaver dissection. Sixteen tacks with a diameter of 1.5 mm and sixteen titanium screws with a diameter of 1 mm were placed into the two wings of the thyroid cartilage. Extrusion forces and the degree of deformation occurring until mechanical failure of the device-body interface were measured for the two types of fixation systems. Results in N and mm were compared using a two sided Wilcoxon test. Neither variable differed significantly between the two groups. However, within the two groups, the necessary strength to pull the tacks or the screws out of the cartilage varied markedly depending on both the inhomogeneous quality of cartilage and the degree of calcification. The PolyMax system with the tacks is recommended as an effective tool for reconstructing the cartilaginous skeleton of the larynx and the trachea with the inherent advantage of resorption as well as avoidance of a second surgery for material removal. PMID- 17123097 TI - Early motor development from partially ordered neural-body dynamics: experiments with a cortico-spinal-musculo-skeletal model. AB - Early human motor development has the nature of spontaneous exploration and boot strap learning, leading to open-ended acquisition of versatile flexible motor skills. Since dexterous motor skills often exploit body-environment dynamics, we formulate the developmental principle as the spontaneous exploration of consistent dynamical patterns of the neural-body-environment system. We propose that partially ordered dynamical patterns emergent from chaotic oscillators coupled through embodiment serve as the core driving mechanism of such exploration. A model of neuro-musculo-skeletal system is constructed capturing essential features of biological systems. It consists of a skeleton, muscles, spindles, tendon organs, spinal circuits, medullar circuits (CPGs), and a basic cortical model. Through a series of experiments with a minimally simple body model, it is shown that the model has the capability of generating partially ordered behavior, a mixture of chaotic exploration and ordered entrained patterns. Models of self-organizing cortical areas for primary somatosensory and motor areas are introduced. They participate in the explorative learning by simultaneously learning and controlling the movement patterns. A scaled up version of the model, a human infant model, is constructed and put through preliminary experiments. Some meaningful motor behavior emerged including rolling over and crawling-like motion. The results show the possibility that a rich variety of meaningful behavior can be discovered and acquired by the neural-body dynamics without pre-defined coordinated control circuits. PMID- 17123098 TI - Tension generation and relaxation in single myofibrils from human atrial and ventricular myocardium. AB - Fast solution switching techniques in single myofibrils offer the opportunity to dissect and directly examine the sarcomeric mechanisms responsible for force generation and relaxation. The feasibility of this approach is tested here in human cardiac myofibrils isolated from small samples of atrial and ventricular tissue. At sarcomere lengths between 2.0 and 2.3 mum, resting tensions were significantly higher in ventricular than in atrial myofibrils. The rate constant of active tension generation after maximal Ca(2+) activation (k (ACT)) was markedly faster in atrial than in ventricular myofibrils. In both myofibril types k (ACT) was the same as the rate of tension redevelopment after mechanical perturbations and decreased significantly by decreasing [Ca(2+)] in the activating solution. Upon sudden Ca(2+) removal, active tension fully relaxed. Relaxation kinetics were (1) much faster in atrial than in ventricular myofibrils, (2) unaffected by bepridil, a drug that increases the affinity of troponin for Ca(2+), and (3) strongly accelerated by small increases in inorganic phosphate concentration. The results indicate that myofibril tension activation and relaxation rates reflect apparent cross-bridge kinetics and their Ca(2+) regulation rather than the rates at which thin filaments are switched on or off by Ca(2+) binding or removal. Myofibrils from human hearts retain intact mechanisms for contraction regulation and tension generation and represent a viable experimental model to investigate function and dysfunction of human cardiac sarcomeres. PMID- 17123099 TI - AtGRP2, a cold-induced nucleo-cytoplasmic RNA-binding protein, has a role in flower and seed development. AB - The glycine-rich protein AtGRP2 is one of the four members of the cold-shock domain (CSD) protein family in Arabidopsis. It is characterized by the presence of a nucleic acid-binding CSD domain, two glycine-rich domains and two CCHC zinc fingers present in nucleic acid-binding proteins. In an attempt to further understand the role of CSD/GRP proteins in plants, we have proceeded to the functional characterization of the AtGRP2 gene. Here, we demonstrate that AtGRP2 is a nucleo-cytoplasmic protein involved in Arabidopsis development with a possible function in cold-response. Expression analysis revealed that the AtGRP2 gene is active in meristematic tissues, being modulated during flower development. Down-regulation of AtGRP2 gene, using gene-silencing techniques resulted in early flowering, altered stamen number and affected seed development. A possible role of AtGRP2 as an RNA chaperone is discussed. PMID- 17123100 TI - Sucrose and starch catabolism in the anther of Lilium during its development: a comparative study among the anther wall, locular fluid and microspore/pollen fractions. AB - In order to better understand the various pathways of sucrose and starch catabolism in the anther of lily (Lilium hybrida var. "Enchantment"), invertase (EC 3.2.1.26) and amylase (EC 3.2.1.1, EC 3.2.1.2) activities were measured separately in different fractions (anther wall, locular fluid and microspore/pollen) and correlated with the sugar content during anther development. Our findings showed significant differences among the fractions analyzed, suggesting that the regulation of sucrose and starch catabolism could follow distinct pathways in each fraction. Glucose and fructose amounts progressively decreased from anther wall to fluid and from fluid to microspore/pollen. Thus, the developing pollen could act as a sink for the carbohydrates that reach the anther. In this sense, cell wall-bound invertases seem to play a major role in soluble sugar partitioning in the different fractions of the anther. Sucrose concentration was found to be substantially higher in the locular fluid than in the other fractions, indicating a probable site for storage. On the other hand, the anther wall tissues could have a buffering function, storing nutrient surplus in starch grains and thus regulating the availability of soluble sugars in the whole anther. All these results proved the advantages of the experimental model proposed here, as well as its usefulness to investigate sugar metabolism in Lilium anthers. PMID- 17123101 TI - Activation of members of a MAPK module in beta-glucan elicitor-mediated non-host resistance of soybean. AB - Plants recognize microbial pathogens by discriminating pathogen-associated molecular patterns from self-structures. We study the non-host disease resistance of soybean (Glycine max L.) to the oomycete, Phytophthora sojae. Soybean senses a specific molecular pattern consisting of a branched heptaglucoside that is present in the oomycetal cell walls. Recognition of this elicitor may be achieved through a beta-glucan-binding protein, which forms part of a proposed receptor complex. Subsequently, soybean mounts a complex defense response, which includes the increase of the cytosolic calcium concentration, the production of reactive oxygen species, and the activation of genes responsible for the synthesis of phytoalexins. We now report the identification of two mitogen-activated protein kinases (MAPKs) and one MAPK kinase (MAPKK) that may function as signaling elements in triggering the resistance response. The use of specific antisera enabled the identification of GmMPKs 3 and 6 whose activity is enhanced within the signaling pathway leading to defense reactions. Elicitor specificity of MAPK activation as well as the sensitivity against inhibitors suggested these kinases as part of the beta-glucan signal transduction pathway. An upstream GmMKK1 was identified based on sequence similarity to other plant MAPKKs and its interaction with the MAPKs was analyzed. Recombinant GmMKK1 interacted predominantly with GmMPK6, with concomitant phosphorylation of the MAPK protein. Moreover, a preferential physical interaction between GmMKK1 and GmMPK6 was demonstrated in yeast. These results suggest a role of a MAPK cascade in mediating beta-glucan signal transduction in soybean, similar to other triggers that activate MAPKs during innate immune responses in plants. PMID- 17123102 TI - Functional identification of sll1383 from Synechocystis sp PCC 6803 as L-myo inositol 1-phosphate phosphatase (EC 3.1.3.25): molecular cloning, expression and characterization. AB - The genome sequence of the cyanobacterium Synechocystis sp. PCC6803 revealed four Open reading frame (ORF) encoding putative inositol monophosphatase or inositol monophosphatase-like proteins. One of the ORFs, sll1383, is approximately 870 base pair long and has been assigned as a probable myo-inositol 1 (or 4) monophosphatase (IMPase; EC 3.1.3.25). IMPase is the second enzyme in the inositol biosynthesis pathway and catalyses the conversion of L-myo-inositol 1 phosphate to free myo-inositol. The present work describes the functional assignment of ORF sll1383 as myo-inositol 1-phosphate phosphatase (IMPase) through molecular cloning, bacterial overexpression, purification and biochemical characterization of the gene product. Affinity (K (m)) of the recombinant protein for the substrate DL-myo-inositol 1-phosphate was found to be much higher (0.0034 +/- 0.0003 mM) compared to IMPase(s) from other sources but in comparison V (max) ( approximately 0.033 mumol Pi/min/mg protein) was low. Li(+) was found to be an inhibitor (IC(50) 6.0 mM) of this enzyme, other monovalent metal ions (e.g. Na(+), K(+) NH (4) (+) ) having no significant effect on the enzyme activity. Like other IMPase(s), the activity of this enzyme was found to be totally Mg(2+) dependent, which can be substituted partially by Mn(2+). However, unlike other IMPase(s), the enzyme is optimally active at approximately 42 degrees C. To the best of our knowledge, sll1383 encoded IMPase has the highest substrate affinity and specificity amongst the known examples from other prokaryotic sources. A possible application of this recombinant protein in the enzymatic coupled assay of L-myo-inositol 1-phosphate synthase (MIPS) is discussed. PMID- 17123103 TI - Coordination of PsAS1 and PsASPG expression controls timing of re-allocated N utilization in hypocotyls of pine seedlings. AB - During pine seed germination, a large amount of N mobilized from the storage proteins is re-allocated in the hypocotyl as free asparagine, as a result of the high levels of asparagine synthetase (AS) encoded by the PsAS1 gene. To determine the role of this re-allocated N reserve, a full-length cDNA encoding L: asparaginase (ASPG) has been cloned from Scots pine (Pinus sylvestris L.) seedlings and characterized. Like other N-terminal nucleophile hydrolases, pine ASPG requires a post-translational processing to exhibit enzymatic activity. However, in contrast to previous reports on other plant ASPGs, purified recombinant pine ASPG does not undergo autoproteolytic cleavage in vitro. Our results suggest that the processing requires accessory proteins to assist in the proteolysis or in the proper folding before autocleavage in a divalent cation dependent manner. Sequence comparison analysis revealed that the pine protein is included in the K+-dependent subfamily of plant ASPGs. The expression of the ASPG encoding gene (PsASPG) was higher in organs with extensive secondary development of the vascular system. The increase in transcript abundance observed at advanced stages of hypocotyl development was concomitant with a decrease of PsAS1 transcript abundance and a remarkable increase in the number of xylem elements and highly lignified cell walls. These results, together with the precise localization of PsASPG transcripts in cells of the cambial region, suggest that the expression of PsAS1 and PsASPG is temporally coordinated, to control the re allocation of N from seed storage proteins toward the hypocotyl to be later used during early development of secondary vascular system. PMID- 17123104 TI - The analysis of the ChlI 1 and ChlI 2 genes using acifluorfen-resistant mutant of Arabidopsis thaliana. AB - One of the key regulatory enzymes of the chlorophyll biosynthesis pathway is magnesium (Mg) chelatase, consisting of three different subunits CHLI, CHLD and CHLH. While CHLH and CHLD are encoded by a single gene each in Arabidopsis, CHLI is encoded by two homologous genes, ChlI 1 and ChlI 2. Analysis of the acifluorfen herbicide resistant mutant aci5 revealed an alteration of the ChlI 1 gene. This mutant as well as wild type plants contained similar transcript levels of the ChlI 1 and ChlI 2 genes. Moreover, the transcripts of both alleles of the ChlI 1 gene were present in the cs (ch42-2)/aci5 hybrid which showed an albina phenotype. Comparison of the amino acid sequence of CHLI 1 and CHLI 2 encoded in the genome of aci5 and wild type revealed in particular alterations of the C terminal end which are suggested to be responsible for the decreased ability of CHLI 2 to participate in the formation of the CHLI ring-like structure of the Mg chelatase complex. PMID- 17123105 TI - Expression of epidermal growth factor receptor (EGFR) is frequent in inflammatory bowel disease (IBD)-associated intestinal cancer. PMID- 17123106 TI - A comparison of pathomolecular markers of fibrosis and morphology in kidney from autopsies of African Americans and whites. AB - African Americans have an increased incidence of chronic kidney disease (CKD) due to hypertension and arteriosclerosis and increased death due to coronary artery disease, compared with whites. The pathogenesis of CKD involves the increased presence and activation of myofibroblasts and macrophages, promotion of tubulointerstitial fibrosis, and effects of tubulointerstitial cell mitosis and apoptosis. We hypothesized that increased risk of hypertensive vascular disease may be identified by renal pathomolecular markers that are associated with progressive CKD. Renal sections were available from 50 autopsies of 33 African Americans (55% males) and 17 whites (76% males) undergoing forensic autopsy for unexpected death. Sclerotic glomeruli, severity of cortical fibrosis, and renal arteriolosclerosis, total glomerular number (N (glom)), average glomerular volume (V (glom)), birth weights, and blood pressure were known. Presence and locality of markers for myofibroblasts (alpha-SMA), macrophages (CD68), collagen, pro fibrotic transforming growth factor-beta1 were scored in renal autopsies, and tubulointerstitial apoptosis was recorded. The results demonstrated a strong positive correlation between age, cortical fibrosis and alpha-SMA (p<0.05), and between CD68 and hypertension and coronary artery disease (p<0.05). The findings confirm the role of myofibroblasts and macrophages in pathogenesis of human CKD. However, the markers showed no significant relationships to V (glom), N (glom), birth weight, or race. PMID- 17123107 TI - P-cadherin and cytokeratin 5: useful adjunct markers to distinguish basal-like ductal carcinomas in situ. AB - Gene expression profiles of invasive breast carcinomas have identified a subgroup of tumours with worse prognosis, which have been called "basal-like". These are characterized by a specific pattern of expression, being estrogen receptor (ER) and HER2 negative, and frequently expressing at least one basal marker such as basal cytokeratins or epidermal growth factor receptor (EGFR). Previously, our group characterized basal-like tumours in a series of invasive breast carcinomas using P-cadherin (P-CD), p63 and cytokeratin 5 (CK5). Based on that study, we hypothesized that those high-grade basal-like invasive carcinomas might have a pre-invasive counterpart, which could be identified using the same approach. A series of 79 ductal carcinomas in situ (DCIS) were classified into distinct subgroups according to their ER, HER2 and basal markers expression. Luminal DCIS expressed ER and constituted 64.6% of the series; the HER2 overexpressing tumours did not express ER and represented 25.3% of the cases, whereas 10.1% lack the expression of ER and HER2 and expressed at least one basal marker (P-CD, CK5, CK14, p63, vimentin and/or EGFR). These basal-like DCIS were mostly high-grade, with comedo-type necrosis, and consistently showed expression of P-CD and CK5. In conclusion, DCIS with a basal-like phenotype represent a small percentage in our series, being P-CD and CK5, the most useful adjunct markers to distinguish this subset of carcinomas in situ of the breast. PMID- 17123108 TI - Cotyledonoid dissecting leiomyoma of the uterus with intravascular growth: report of two cases. AB - We present two cases of cotyledonoid dissecting leiomyoma of the uterus with intravascular involvement, which occurred in women aged 73 and 48 years. Grossly and microscopically, both neoplasms had an extrauterine cotyledonoid part and intrauterine dissecting fascicles of disorganized, swirled neoplastic smooth muscle with hydropic degeneration and foci of an intravascular growth (the latter was identified histologically). To our knowledge, the intravascular component of such a neoplasm is a very rare feature that has previously been described only in three cases in the literature. PMID- 17123109 TI - Insulinoma misdiagnosed as juvenile myoclonic epilepsy. PMID- 17123110 TI - Twelve cases of tinea unguium in a pediatric clinic in 9 years. AB - Tinea of the nails is not an exclusively adult pathology. The pediatrician should include this entity in the differential diagnosis. PMID- 17123111 TI - Wheat VIN3-like PHD finger genes are up-regulated by vernalization. AB - The term 'vernalization' describes the acceleration of the transition between the vegetative and reproductive stages after exposing plants to an extended period of low temperature. In Arabidopsis, vernalization promotes flowering by silencing the flowering repressor gene FLOWERING LOCUS C (FLC). Mitotically stable repression of FLC is the result of chromatin modifications mediated by the Vernalization-INsensitive 3 (VIN3) and VIN3-Like (VIL) proteins. In this study, we identified and characterized three VIL genes in diploid wheat (Triticum monococcum L.), named TmVIL1, TmVIL2, and TmVIL3. Similar to Arabidopsis VIN3, all three wheat VIL proteins carry three conserved domains including a plant homeodomain finger motif (PHD), a fibronectin type III domain (FNIII), and a VIN3 interacting domain (VID). Genetic mapping placed TmVIL1, TmVIL2, and TmVIL3 loci in the centromeric regions of chromosome 5, 6, and 1, respectively. The chromosome location of TmVIL1 is close to that of the vernalization gene VRN-D5, but more precise mapping information is required to validate this relationship. Transcription of the wheat VIL genes was up-regulated by vernalization, with a peak after 4-6 weeks of cold treatment. When transferred back to warm conditions, transcript levels of the wheat VIL genes returned to pre-vernalization levels. In addition, the transcript levels of wheat VIL genes are affected by photoperiod. This study indicates that wheat VIL genes have retained a similar structure and transcriptional regulation as their Arabidopsis VIN3/VIL homologues, suggesting that they might have retained some of their functions. PMID- 17123112 TI - Ecological role of reindeer summer browsing in the mountain birch (Betula pubescens ssp. czerepanovii) forests: effects on plant defense, litter decomposition, and soil nutrient cycling. AB - Mammalian herbivores commonly alter the concentrations of secondary compounds in plants and, by this mechanism, have indirect effects on litter decomposition and soil carbon and nutrient cycling. In northernmost Fennoscandia, the subarctic mountain birch (Betula pubescens ssp. czerepanovii) forests are important pasture for the semidomestic reindeer (Rangifer tarandus). In the summer ranges, mountain birches are intensively browsed, whereas in the winter ranges, reindeer feed on ground lichens, and the mountain birches remain intact. We analyzed the effect of summer browsing on the concentrations of secondary substances, litter decomposition, and soil nutrient pools in areas that had been separated as summer or winter ranges for at least 20 years, and we predicted that summer browsing may reduce levels of secondary compounds in the mountain birch and, by this mechanism, have an indirect effect on the decomposition of mountain birch leaf litter and soil nutrient cycling. The effect of browsing on the concentration of secondary substances in the mountain birch leaves varied between different years and management districts, but in some cases, the concentration of condensed tannins was lower in the summer than in the winter ranges. In a reciprocal litter decomposition trial, both litter origin and emplacement significantly affected the litter decomposition rate. Decomposition rates were faster for the litter originating from and placed into the summer range. Soil inorganic nitrogen (N) concentrations were higher in the summer than in the winter ranges, which indicates that reindeer summer browsing may enhance the soil nutrient cycling. There was a tight inverse relationship between soil N and foliar tannin concentrations in the winter range but not in the summer range. This suggests that in these strongly nutrient-limited ecosystems, soil N availability regulates the patterns of resource allocation to condensed tannins in the absence but not in the presence of browsing. PMID- 17123113 TI - Bias and precision of estimated glomerular filtration rate in children. AB - Determining true glomerular filtration rate (GFR) using an exogenous marker is time-consuming and cumbersome. Therefore, creatinine-based estimates of GFR are used. Recent papers using new population-specific/local parameters in their prediction equations, standardizing creatinine determination or adding other endogenous surrogate markers of GFR, like cystatin C, could demonstrate an improvement of bias inherent in the results of the prediction equations. Precision, however, is still poor. Currently, we have to accept a precision (as defined in the so-called Bland-Altman plot) of +/-20% in adults and +/-30-40% in children. This problem of poor precision/uncertainty is especially bothering in the higher, near normal GFR range. Caution should be exercised when applying prediction equations in individuals in need of an accurate GFR determination. In that case, a real clearance procedure has to be performed. In the long run, the true clearance procedure should be simplified using new exogenous GFR markers and developing new devices, allowing GFR measurements to be performed, for example, transcutaneously. Such a procedure would be more acceptable for both patients and physicians. PMID- 17123114 TI - Percutaneous translumbar inferior vena cava catheter placement for long-term hemodialysis treatment. AB - Central venous access for long term dialysis has always been a major difficulty, especially in cases where the traditional sites are no longer available. We present a case of a pediatric patient where the usual sites were occluded and he needed a reliable venous access for his dialysis. The inferior vena cava has been used in adult patients, but no reports were found in the pediatric literature. We chose this site as an alternative for this patient. The catheter was placed via a percutaneous translumbar approach, and remained in place for almost 2 years until the patient received a renal transplant. PMID- 17123115 TI - Comparison of ambulatory blood pressure and Task Force criteria to identify pediatric hypertension. AB - The aim of this study was to assess the level of agreement between central European ambulatory blood pressure monitoring (ABPM) and American Task Force (TF) criteria when applied to blood pressure (BP) measurements collected by ABPM to evaluate patients with hypertension. In 169 patients, we applied both sets of criteria and calculated mean daytime and nighttime BP and daytime BP loads. The frequency of hypertension for daytime systolic BP was significantly higher when TF criteria were used (p or=50% were evaluated (p or=10% WG prior to PICU admission compared with 15 (22.1%) in the non-PICU group (p<0.001). Thirteen patients (68.4%) developed>or=10% FO prior to PICU admission compared with 31 (45.6%) in the non-PICU group (p=0.075). Following multivariate analysis, >or=10% WG (p=0.018) and cardiac dysfunction on admission for HSCT (p=0.036) remained independent risk factors for PICU admission. Smaller children (p=0.033) and patients with a twofold increase in serum creatinine (p=0.026) were at risk of developing>or=10% WG. This study shows that WG is a risk factor for PICU admission in pediatric HSCT recipients. Further research is needed to better understand the pathophysiology of WG in these patients and to determine the impact of WG prevention on PICU admission. PMID- 17123120 TI - Tacrolimus-induced HUS: an unusual cause of acute renal failure in nephrotic syndrome. AB - Acute renal failure (ARF) is an uncommon complication in children with nephrotic syndrome. We report here the case of a 10-year-old male child with primary steroid-resistant nephrotic syndrome who was non-responsive to steroids and cyclophosphamide. A kidney biopsy revealed that he had focal segmental glomerulosclerosis. His treatment was initiated with tacrolimus (dose of 0.15 mg/kg/day) in two divided doses along with prednisolone 60 mg/m(2)/daily. After 1 month of treatment, he was diagnosed as having acute renal failure secondary to HUS. This was postulated to be due to the tacrolimus therapy, which was withdrawn. Two weeks after stopping the adminsitration of tacrolimus, his urine output improved, and the hemoglobin and serum creatinine normalized. Thus, tacrolimus-induced HUS is a rare cause of ARF in nephrotic syndrome. With the increasing use of tacrolimus in steroid-resistant nephrotic syndrome, the treating physicians need to be aware of this rare, but potentially life threatening side effect. PMID- 17123121 TI - [Compulsive shopping--current considerations on classification and therapy]. AB - Compulsive shopping is classified by ICD-10 (F63.8) as an "impulse control disorder, not otherwise classified". Several authors consider compulsive shopping rather as a variety of dependence disorder. It is characterized by the impulsive or compulsive buying of unneeded things, personal distress, impaired social and vocational functioning, and/or financial problems. In this case, we discuss a two way therapy consisting of addiction-specific psychological education and high dose selective serotonin reuptake inhibitors (SSRIs). We further point to compliance problems caused by SSRI side effects. PMID- 17123122 TI - [Neuropsychological aspects of delusional disorders. Characteristic attributional style or cognitive deficit?]. AB - BACKGROUND: "Pure" delusional disorders are clinically rare, and the neuropsychology of such disorders is poorly understood. Whereas "deficit" models suggest a cognitive impairment accounting for the incorrigible fixation of false beliefs, cognitive models propose the existence of a characteristic attributional style in patients to stabilise a fragile self. PATIENTS AND METHODS: The cognitive flexibility and attributional style of 21 patients diagnosed with delusional disorder according to ICD-10 were compared with a group of healthy controls paralleled for age, sex, education, and intelligence. RESULTS: Patients with delusional disorders made more errors and more perseverative errors in the Wisconsin Card Sorting Test compared with controls. However, these differences were only significant in patients with a comorbid depression. In contrast to earlier studies, patients with delusional disorders did not attribute negative events to external or personal causes more often than healthy controls, but partly tended to show a depressive attributional style. CONCLUSION: Our results do not support either a cognitive deficit in patients with delusional disorders or a characteristic attributional style. In terms of treatment recommendations, a thorough diagnosis of comorbid depressive disorders in patients with delusional disorders is warranted. PMID- 17123123 TI - [Acute and transient psychotic disorders (ICD-10: F23). Empirical data and implications for therapy]. AB - The defining clinical features of acute and transient psychotic disorders (ICD 10: F23) are an acute onset and a duration of psychotic symptoms not exceeding 1 3 months. Previous empirical investigations show that patients with this diagnosis have a favourable prognosis, but also a high risk of relapses (58-77%). The diagnostic stability in the further course of illness seems to be low (34 73%) with a frequent diagnostic change to schizophrenia and affective disorders being observed. As yet, data from controlled clinical trials regarding therapy do not exist. The implications for the treatment of acute and transient psychotic disorders are discussed with this background. PMID- 17123124 TI - [The brain-mind debate. Problems in the philosophy of science with regard to psychiatry]. AB - The present interdisciplinary brain-mind debate with regard to neurobiology shows deficits in the criticism of methods and in the precision of language and argumentation. Simplifying localisations of psychic functions, insufficient explanations and over-interpretations in the sense of physical determinism are the consequence. This can be demonstrated in deficits of neurobiological theories of volitional action. Therefore, the current concept of man is less perturbed than is proposed by neurobiologists. For psychiatry besides the neurobiological approach also a separate way of building models seems to be useful if present theoretical psychology and systems science would be regarded. In particular, the systems science of natural systems offers new opportunities to bridge gaps between disciplines involved in the brain-mind debate. PMID- 17123125 TI - Parent satisfaction in a multi-site acute trial of risperidone in children with autism: a social validity study. AB - RATIONALE: Subjects who view experimental procedures as worthwhile are more likely to participate in clinical trials and comply with study procedures. Designing studies that consider the consumer's perspective will help to forge a better alliance between participants and researchers. OBJECTIVE: Participant satisfaction is seldom assessed in pharmacological research. In this paper, we report on parent satisfaction in a randomized clinical trial in children with autistic disorder and severely disruptive behavior. METHOD: Parents of 101 children with autism who had participated in a multi-site 8-week double-blind clinical trial of risperidone were given a questionnaire at the end to elicit their perceptions of the appropriateness and acceptability of clinical trial procedures. RESULTS: Ninety-six (95.0%) parents returned the questionnaire. Of these, 80.0 to 96.8%, depending on the question, expressed satisfaction with their child's research participation regardless of treatment outcome or assignment to active drug or placebo. In all, 90.5% of parents indicated that they would "definitely" recommend the clinical trial to other families with similar children. A total of 92.7% indicated that they would rejoin the clinical trial if they had to do it all over again. Ethnic minority subjects were more satisfied than white participants with the use of "learning tests". CONCLUSIONS: Parents of children participating in this trial were highly satisfied and supportive of the clinical trial procedures. Random assignment to drug or placebo and the clinical response of their children did not appear to influence their views. Further satisfaction studies of this sort are encouraged. PMID- 17123126 TI - Germ-cell cluster formation in the telotrophic meroistic ovary of Tribolium castaneum (Coleoptera, Polyphaga, Tenebrionidae) and its implication on insect phylogeny. AB - Tribolium castaneum has telotrophic meroistic ovarioles of the Polyphaga type. During larval stages, germ cells multiply in a first mitotic cycle forming many small, irregularly branched germ-cell clusters which colonize between the anterior and posterior somatic tissues in each ovariole. Because germ-cell multiplication is accompanied by cluster splitting, we assume a very low number of germ cells per ovariole at the beginning of ovariole development. In the late larval and early pupal stages, we found programmed cell death of germ-cell clusters that are located in anterior and middle regions of the ovarioles. Only those clusters survive that rest on posterior somatic tissue. The germ cells that are in direct contact with posterior somatic cells transform into morphologically distinct pro-oocytes. Intercellular bridges interconnecting pro-oocytes are located posteriorly and are filled with fusomes that regularly fuse to form polyfusomes. Intercellular bridges connecting pro-oocytes to pro-nurse cells are always positioned anteriorly and contain small fusomal plugs. During pupal stages, a second wave of metasynchronous mitoses is initiated by the pro-oocytes, leading to linear subclusters with few bifurcations. We assume that the pro oocytes together with posterior somatic cells build the center of determination and differentiation of germ cells throughout the larval, pupal, and adult stages. The early developmental pattern of germ-cell multiplication is highly similar to the events known from the telotrophic ovary of the Sialis type. We conclude that among the common ancestors of Neuropterida and Coleoptera, a telotrophic meroistic ovary of the Sialis type evolved, which still exists in Sialidae, Raphidioptera, and a myxophagan Coleoptera family, the Hydroscaphidae. Consequently, the telotrophic ovary of the Polyphaga type evolved from the Sialis type. PMID- 17123127 TI - Thermal stability and biochemical properties of isocitrate dehydrogenase from the thermoacidophilic archaeon Thermoplasma acidophilum. AB - Isocitrate dehydrogenase [IDH; EC 1.1.1.42] from the thermoacidophilic archaeon Thermoplasma acidophilum (TaIDH) showed high thermal stability with an apparent melting temperature, T(m), of 82.2 and 84.5 degrees C at pH 7.5 and 5.8, respectively. Based on structural alignment of TaIDH with IDH from Aeropyrum pernix (ApIDH) and Archaeoglobus fulgidus (AfIDH) residues forming an aromatic cluster in the clasp-domain thought to strengthen the dimer interface in ApIDH and AfIDH were identified in the former enzyme. Moreover, TaIDH had a shortened N terminus that may protect the enzyme from thermal denaturation. The enzyme activity of TaIDH was highest at 70 degrees C. The pH-activity profile was bell shaped with an optimum shifted to a lower pH compared to AfIDH. The activity of TaIDH was influenced by changes in pH with a three-fold reduction in activity when the pH was shifted from the pH-optimum at 7.5 to pH 5.8. However, the specific activity at pH 5.8 was still high when compared with AfIDH. The reduction in activity at pH 5.8 was not due to instability of the enzyme as the T(m) of TaIDH was higher at pH 5.8 than at 7.5 and the enzyme retained 91% of its activity after incubation at 1 h at pH 5 and 60 degrees C. The difference in the pH-profile of TaIDH in comparison with AfIDH may thus be related to the pK(a)s of their catalytic residues involved in the initial proton abstraction and the final proton donation during the catalysis of oxidative decarboxylation of isocitrate to 2-oxoglutarate and reduced coenzyme. PMID- 17123128 TI - Proteomic analysis of Psychrobacter cryohalolentis K5 during growth at subzero temperatures. AB - It is crucial to examine the physiological processes of psychrophiles at temperatures below 4 degrees C, particularly to facilitate extrapolation of laboratory results to in situ activity. Using two dimensional electrophoresis, we examined patterns of protein abundance during growth at 16, 4, and -4 degrees C of the eurypsychrophile Psychrobacter cryohalolentis K5 and report the first identification of cold inducible proteins (CIPs) present during growth at subzero temperatures. Growth temperature substantially reprogrammed the proteome; the relative abundance of 303 of the 618 protein spots detected (approximately 31% of the proteins at each growth temperature) varied significantly with temperature. Five CIPs were detected specifically at -4 degrees C; their identities (AtpF, EF Ts, TolC, Pcryo_1988, and FecA) suggested specific stress on energy production, protein synthesis, and transport during growth at subzero temperatures. The need for continual relief of low-temperature stress on these cellular processes was confirmed via identification of 22 additional CIPs whose abundance increased during growth at -4 degrees C (relative to higher temperatures). Our data suggested that iron may be limiting during growth at subzero temperatures and that a cold-adapted allele was employed at -4 degrees C for transport of iron. In summary, these data suggest that low-temperature stresses continue to intensify as growth temperatures decrease to -4 degrees C. PMID- 17123129 TI - The lysogenic region of virus phiCh1: identification of a repressor-operator system and determination of its activity in halophilic Archaea. AB - phiCh1 is a temperate virus infecting the haloalkaliphilic archaeon Natrialba magadii. As for all temperate viruses, a control of the lysogenic state versus the lytic life cycle is essential. Two open reading frames (ORFs) have been identified as putative repressor encoding genes: ORF48 and ORF49. The protein of ORF48 showed sequence similarities to putative repressor molecules. ORF49 was identified by the analysis of a mutant of phiCh1: the lysogenic strain carrying mutant phiCh1-1 showed a different lysis behavior than wild type virus phiCh1, indicating a dysfunction in the regulation of gene expression. Here, we show that the intergenic region between ORF48 and ORF49 comprises a promoter/operator sequence that is a transcriptionally active region in the model system Haloferax volcanii. Transcription from this region can be repressed by the activity of the ORF48 gene product. Gp43/gp44 has an enhancing effect on this regulatory sequence. Evidence is given for a possible binding site of Rep and gp43/gp44 within the coding region of the rep gene. PMID- 17123130 TI - Prognostic model of stage II non-small cell lung cancer by a discriminant analysis of the immunohistochemical protein expression. AB - PURPOSE: We aimed to identify the key proteins that influence the prognosis of non-small cell lung cancer (NSCLC) using protein expression profiles of previously known prognostic markers. METHODS: Thirty-one cases of Stage II NSCLC with 5-year follow-up data were selected. Tissue microarrays (TMA) and immunohistochemistry were used to make protein expression profiles of 18 previously reported immunohistochemical prognostic markers and their value in NSCLC was statistically re-evaluated by a discriminant analysis. RESULTS: For the discriminant analysis using marker protein expression profiles, we selected three significant markers, TTF-1, RCAS1 and c-MET, to evaluate each patient's 5-year survival. The requested discriminant function was V = -1.08754 x (RCAS1 score) - 0.83174 x (TTF1 score) + 0.55204 x (cMET score) + 5.46972, and V = 0 served as a cut-off point. The correctness for evaluating a patient's 5-year survival by a discriminant analysis was 87.1%. CONCLUSIONS: A discriminant analysis is thus considered to be a useful statistical method for analyzing the protein expression profiles obtained by combined TMA and immunohistochemical techniques using archival NSCLC tissues. However, the sample size and selection of the marker protein depending on the histology greatly influence the results of a NSCLC study. PMID- 17123131 TI - Re-evaluation of axillary skip metastases in the era of sentinel lymph node biopsy in breast cancer. AB - PURPOSE: To investigate whether skip axillary metastases are really skip metastases or a continuation of level I micrometastases in invasive breast cancer, and to determine whether there are any factors predisposing to skip metastases. METHODS: We reviewed 568 consecutive patients with breast cancer who underwent complete axillary lymph node dissections (ALND) between January 1998 and December 2004. For patients with skip axillary lymph node metastases, resectioning and immunohistochemical staining of the remaining part of paraffin blocks from level I lymph nodes were done to determine whether there were any micrometastases in this group of lymph nodes. RESULTS: Skip axillary metastases were found in 27 (10%) of 268 patients with axillary lymph node metastases. Re evaluation of the level I lymph nodes, both with thin sectioning and immunohistochemical staining, in the patients with axillary skip metastases revealed no micrometastases. No significant correlation was found between the demographic and histopathological variables of the patients with skip metastases and those with regular axillary metastases. CONCLUSIONS: These results suggest that skip axillary metastases are actual skip metastases, not a continuation of undetected level I micrometastases. Moreover, none of the clinical and histopathological measures of primary tumors are predictors of the presence of skip metastases. PMID- 17123132 TI - Factors associated with postoperative cognitive dysfunction in patients undergoing cardiac surgery. AB - PURPOSE: Central nervous system complications continue to be major causes of morbidity and mortality after cardiac surgery. The purpose of this study was to identify the risk factors for postoperative cognitive dysfunction after coronary artery bypass graft (CABG) surgery. METHODS: Eighty-eight patients scheduled for elective CABG were studied. After the induction of anesthesia, a fiberoptic oximetry oxygen saturation catheter was inserted into the right jugular bulb for the continuous monitoring of jugular venous oxygen hemoglobin saturation (SjvO(2)). The hemodynamic parameters and arterial and jugular venous blood gases were measured during cardiopulmonary bypass (CPB). All patients underwent a battery of neurological and neuropsychological tests one day before the operation and at 6 months after the operation. RESULTS: The incidence of a cognitive decline at 6 months was 24/88 (27.3%). Greater age (P = 0.04), the presence of renal failure (P < 0.001), and diabetes mellitus (P < 0.001) were more frequent in the patients with postoperative cognitive dysfunction at 6 months after the operation than in patients without cognitive dysfunction. Age (odds ratio [OR], 1.4; 95% confidence interval [CI], 1.0-1.7; P = 0.04), diabetes mellitus (OR, 1.8; 95% CI, 1.2-2.4; P < 0.01), and presence of renal failure (OR, 2.8; 95% CI, 2.4-4.3; P < 0.01) were associated with cognitive impairment at 6 months postoperatively. However, there was no relationship between the presence of atherosclerosis in the ascending aorta and postoperative cognitive dysfunction after CABG surgery. CONCLUSIONS: A greater age, diabetes mellitus, and renal failure were found to be risk factors for development of cognitive impairment at 6 months after CABG with CPB. PMID- 17123133 TI - Long-term outcome after hernia repair with the prolene hernia system. AB - PURPOSE: We evaluated the long-term outcome and symptoms after inguinal hernia repair using the Prolene Hernia System (PHS). METHODS: We reviewed the medical records of all patients who underwent PHS surgery at our hospital to assess the postoperative complications and recurrence rates. Late symptoms were evaluated by a standardized interview using a questionnaire. RESULTS: A total of 395 hernias in 367 patients were studied. Postoperative complications developed after 20 repairs (5.1%), including a severe mesh infection in one patient. Seven (1.8%) patients suffered recurrence. Of 395 cases, 363 (91.9%) responded to the interview. After a median follow-up interval of 19.3 months (range 1.0-55.8), moderate pain and moderate discomfort were reported by only 1.9% and 0.8% of patients, respectively. CONCLUSIONS: Prolene Hernia System surgery is a safe and feasible procedure with low rates of complication, recurrence, and late symptoms. PMID- 17123134 TI - Favorable effects of preoperative enteral immunonutrition on a surgical site infection in patients with colorectal cancer without malnutrition. AB - PURPOSE: A prospective study was conducted to ascertain the effects of preoperative enteral immunonutrition on a surgical site infection (SSI) in patients with colorectal cancer without malnutrition. METHODS: Patients with colorectal cancer undergoing elective surgery without malnutrition, bowel obstruction, severe cardiopulmonary complications, diabetes, collagen disease, or renal failure were sequentially divided into an immunonutrition group (n = 33) and a control group (n = 34). In the immunonutrition group, an enteral diet supplemented with arginine, dietary nucleotides, and omega-3 fatty acids was administered for 5 days (750 ml/day) prior to surgery. RESULTS: The mean age was slightly higher in the immunonutrition group (69 +/- 9 years) than in the control group (63 +/- 11 years; P < 0.05), but no significant differences between the groups were noted for the body mass index, total protein, albumin, hemoglobin, surgical methods, operation time, or volume of intraoperative bleeding. The frequencies of superficial incisional SSI, deep incisional SSI and organ/space SSI in the immunonutrition and control groups were 0% and 11.8% (4/34; P < 0.05), 0% and 0%, and 0% and 2.9% (1/34), respectively. CONCLUSION: Preoperative enteral immunonutrition appears to be effective for preventing SSI in patients with colorectal cancer without malnutrition. PMID- 17123135 TI - Synergistic antitumor effect of an angiogenesis inhibitor (TNP-470) and tumor necrosis factor in mice. AB - PURPOSE: We investigated the potentiation of combination therapy using tumor necrosis factor (TNF) with TNP-470, a potent angiogenesis inhibitor. METHODS: We evaluated the antitumor effect in vivo against subcutaneous (s.c.) MC38 mouse colon adenocarcinoma tumors in C57BL/6 mice. The mice were treated with a single bolus injection via the tail vein of 3 or 8 microg rhTNF in 0.5% bovine serum albumin/normal saline (BSA/NS), or with 0.5% BSA/NS alone as a control, with or without TNP-470 pretreatment, given as 30 or 60 mg/kg x 2 days, s.c. DNA synthesis in human umbilical endothelial cells (HUVEC) was assessed by [(3)H]thymidine uptake after incubation with TNF, with or without TNP-470. RESULTS: The antitumor effect of TNP-470 pretreatment combined with 3 microg recombinant human (rh) TNF injection resulted in an 80% reduction of tumor volume compared with the control. This was significantly better than that induced by 3 microg rhTNF alone (P < 0.005). DNA synthesis in HUVEC was inhibited by TNF with TNP-470 in a dose-dependent manner, but there was no enhanced effect against MC38 in vitro. CONCLUSIONS: These results suggest that the combination of the angiogenesis inhibitor TNP-470 and TNF might have a synergistic antitumor effect on solid tumors in vivo. PMID- 17123136 TI - Tumor viability using real-time spectral images. AB - PURPOSE: We observed real-time spectral images using line-scan imaging spectrography in order to evaluate tumor viability. METHODS: Japanese albino rabbits, which underwent the implantation of VX2 tumor strain, were used as subcutaneous tumor models (n = 54). Photodynamic therapy (PDT) consisted of semiconductor laser irradiation of the tumorous lesion. The experimental groups consisted of a PDT group (n = 15) and a non-PDT group (control group, n = 15). The spectral images taken by a CCD camera underwent computer processing. Next, the spectrum of the tumorous lesion was observed and the peak spectrum value was measured. RESULTS: Two peaks (545 and 575 nm) corresponding to the absorbance spectrum of oxygenated hemoglobin (oxyHgb) were observed in the untreated area. In the treated area, however, they disappeared and a different peak (560 nm) corresponding to the absorbance spectrum of deoxygenated hemoglobin was observed. PDT induced ischemic tissues and the cells could be confirmed in real time in vivo in monochrome and color images reflecting the oxyHgb amount. A histopathological examination and phosphotungstic acid hematoxylin staining demonstrated diffuse fibrin accumulation in the microvessels, while proliferating cell nuclear antigen staining showed a reduced nuclear stainability. CONCLUSIONS: These results demonstrated that the real-time spectral images showed the actual histological conditions such as a blocked tumor blood flow and reduced tumor viability. PMID- 17123137 TI - Protein pattern difference in the colon cancer cell lines examined by two dimensional differential in-gel electrophoresis and mass spectrometry. AB - PURPOSE: The pivotal metastatic processes of colorectal cancer (CRC) have yet to be fully investigated by a comprehensive all-inclusive protein analysis. We used two-dimensional differential in-gel electrophoresis (2D-DIGE) and liquid chromatography-tandem mass spectrometry (LC/MS/MS) to investigate the protein pattern changes during the metastasis of CRC. Two CRC cell lines were investigated: SW480 derived from the primary lesion and SW620 derived from lymph node metastasis in the same patient. METHODS: The two cell lines were compared using 2D-DIGE with a maleimide CyDye fluorescent protein labeling technique, which has an enhanced sensitivity for many proteins at a low concentration. A comprehensive proteomics analysis was performed by the dual-labeling method using Cy3 and Cy5 and by LC/MS/MS. In addition, an in vivo experiment of metastasis using nude mice was performed by the injection of the two cell lines into the spleen. RESULTS: Among approximately 1,500 proteins, we detected 9 protein spots with definitively significant changes between the two cell lines. Three out of the nine proteins were validated by a Western blot analysis. Alpha-enolase and triosephosphate isomerase were significantly upregulated in SW620 in comparison to SW480. Annexin A2 (annexin II) was significantly downregulated in SW620 compared to SW480. Neither liver metastasis nor peritoneal dissemination was established in the metastatic experiment using SW480 but some liver and peritoneal metastases occurred in the experiment using SW620. An in vivo metastatic experiment using SW620 showed the expressions of alpha-enolase and triosephosphate isomerase to increase in the liver metastases in comparison to those in the splenic implanted lesion. The expressions of triosephosphate isomerase increased in the peritoneal lesions in comparison to those in the splenic implanted lesion. CONCLUSIONS: 2D-DIGE and LC/MS/MS techniques identified nine proteins that increased significantly more in SW620 than in SW480. The finding of our in vivo metastatic experiment suggests that alpha-enolase and triosephosphate isomerase, at least in part, may be associated with the metastatic process of these two cell lines. PMID- 17123138 TI - Splenic autotransplantation for a congested and enlarged wandering spleen with torsion: report of a case. AB - In children with diseases of the spleen, every effort should be made to preserve the organ, to prevent severe infections postsplenectomy. We report the case of a 7-year-old girl with torsion of a wandering spleen who we treated by autotransplantation of splenic tissues following splenectomy, when fixation of the enlarged spleen seemed impossible. Spleen scintigraphy showed uptake in the regenerating splenic tissues 9 months after surgery, and evidence of an increase in the size of the tissues 23 months after surgery. Howell-Jolly bodies had disappeared by 16 months after surgery. These findings suggested that the transplanted splenic tissues were resuming splenic functions. Based on our experience with this case, we conclude that autotransplantation after splenectomy is a treatment option for wandering spleen with torsion when fixation seems difficult because of splenic congestion and enlargement. PMID- 17123139 TI - Adventitial cystic disease of the popliteal vein: report of a case. AB - Adventitial cystic disease (ACD), also known as cystic mucoid or myxomatous degeneration, is a rare vascular disease seen mainly in arteries. It is very unusual for these cystic masses to develop in a vein. We report the case of a 56 year-old woman with leg swelling caused by ACD arising in the popliteal vein. The swelling appeared after a long period of standing. Magnetic resonance imaging (MRI) showed a popliteal cystic mass and venography showed disrupted venous flow. We resected the cyst wall without venous reconstruction, after which venous blood flow normalized and her symptoms subsided. To our knowledge, this is only the third documented case of ACD arising in the popliteal vein. A misdiagnosis could easily have been made, since the mass was not obvious on physical examination and the only symptom was intermittent swelling. Thus, it is important to be aware of ACD as a possible diagnosis when examining patients with a swelling in the leg. PMID- 17123140 TI - Postradiation sarcoma of the chest wall: report of two cases. AB - Postradiation sarcoma is a rare late complication of external radiotherapy. We herein present two cases with this disease. A 54-year-old man had undergone a lobectomy and chest wall resection for Pancoast type lung cancer 7 years previously. He had undergone irradiation with a total dose of 50 Gy. Computed tomography (CT) demonstrated a tumorous expansion of the right lateral thoracic wall. A pathological examination confirmed a diagnosis of osteosarcoma. A 60-year old woman had undergone a resection of the lateral chest wall mass, which was diagnosed to be Hodgkin's disease in 1991. Chemotherapy was given postoperatively. A tumorous lesion arose again and irradiation was performed with a total dose of 110 Gy. In 2000, two tumors appeared in the irradiation field. A pathological examination showed a sarcoma with divergent differentiation. In 2003, a tumor recurred and was diagnosed to be a liposarcoma. Patients who have received radiotherapy should therefore be followed up while taking into consideration the possible development of postradiation sarcoma. PMID- 17123141 TI - Medullary carcinoma of the breast after radiation therapy for Hodgkin's disease in a young woman: report of a case. AB - A 28-year-old woman who had received mantle-field radiation therapy for Hodgkin's disease (HD) almost 10 years earlier was referred to our hospital for investigation of a palpable mass in her left breast. Mammography, sonography, and fine-needle aspiration were done and the results showed medullary carcinoma of the breast, an uncommon type of breast cancer in young women. We review the literature on the secondary development of breast cancer after radiation therapy for HD. PMID- 17123142 TI - Noradrenalin-secreting retroperitoneal schwannoma resected by hand-assisted laparoscopic surgery: report of a case. AB - A 66-year-old woman was admitted to our hospital for investigation of a retroperitoneal mass. She was asymptomatic but her serum noradrenalin and vanillylmandellic acid levels were increased remarkably. Computed tomographic angiography showed a mass in the left side of the superior mesenteric artery (SMA) divergence, 6 x 4 cm in size, supplied mainly by the left adrenal artery. We performed hand-assisted laparoscopic surgery (HALS) using a Cavitron ultrasonic surgical aspirator. Abdominal exploration revealed that the tumor was located in the left side of the SMA root, but was not adhered to the adjacent organs. Her vital signs remained stable during the operation. Microscopic examination revealed a palisade formation, confirming a histopathological diagnosis of degenerated schwannoma (Antoni type B). Immunohistologically, the tumor cells were stained with S-100 protein and neuron-specific enolase. Her noradrenalin level normalized immediately after the operation and she remains in good health 3 years 2 months later. PMID- 17123143 TI - Spontaneous perforation of Meckel's diverticulum without peritonitis in a newborn: report of a case. AB - A case of a newborn male with a perforation of Meckel's diverticulum is reported. The clinical course consisted of progressive abdominal distention and pneumoperitoneum that formed within 29 h after birth. The perforation of Meckel's diverticulum was not associated with peritonitis because meconium did not contaminate the abdominal cavity. The histology of the diverticulum showed a nearly intact muscular layer but a focal muscular defect. Neither any inflammatory phenomena nor ectopic mucosa was found. A congenital focal muscular defect of the diverticulum and a sudden elevation of intraluminal pressure due to bowel movement after birth may thus be the pathogenesis of a spontaneous perforation. A search of the English literature did not reveal any similar case. PMID- 17123144 TI - Bleeding Meckel diverticulum associated with a vitellointestinal artery aneurysm found on preoperative angiography: report of a case. AB - An 18-year-old man was admitted to a local hospital with abdominal pain and bloody stool. Upper and lower gastrointestinal endoscopy failed to show any bleeding sites; however, an angiography of the superior mesenteric artery done on hospital day 4 showed an abnormal artery with an aneurysm, branching from the ileal artery. This artery was thought to be the vitellointestinal artery, a feeding artery of Meckel diverticulum. After embolization, he was transferred to our hospital, where we performed emergency laparotomy with partial resection of the ileum, including a bleeding Meckel diverticulum. Pathological examination revealed ectopic gastric mucosa and peptic ulceration, which we assumed was the origin of the bleeding. The patient had an uneventful postoperative course. Visceral artery aneurysms are rare but important vascular lesions because of their potential for fatal rupture. Although a minimally invasive procedure can be performed for a vitellointestinal artery aneurysm in patients with asymptomatic Meckel diverticulum, we treated our patient surgically because he presented with hemorrhagic shock and had been unresponsive to an H(2)-receptor antagonist. PMID- 17123145 TI - Compartment syndrome of bilateral lower extremities following laparoscopic surgery of rectal cancer in lithotomy position: report of a case. AB - A 67-year-old man underwent laparoscopic surgery for rectal cancer in the lithotomy position. After surgery he complained of bilateral lower limb pain, swollen legs, and sensory disturbance. The serum creatine kinase value was 46 662 U/l. Venography demonstrated compression from outside without any obstruction. The T2 image of magnetic resonance imaging (MRI) showed a massive swollen muscle and a partial high-intensity area in the bilateral lower limbs. The posterior compartment pressures of lower legs were high (gastrocnemius muscle: 30 mmHg [right] and 44 mmHg [left]). Compartment syndrome (superficial posterior compartment) was thus diagnosed. He underwent a fasciotomy using the single dorsal approach and the administration of a large amount of fluid. He recovered well without any motor or sensory deficits. Compartment syndrome is rare, occurring only once in every 3500 cases, but it is a severe complication of surgery in the lithotomy position. Several risk factors have been pointed out: including prolonged operation, hardness of the operating table, obesity, dehydration, and hypothermia. To prevent compartment syndrome, appropriate positioning during surgery is therefore essential. To make a timely diagnosis and identify the precise location of muscle edema, the T2 image of MRI is useful. PMID- 17123146 TI - Atresia of the ileocecal junction with agenesis of the ileocecal valve and vermiform appendix: report of a case. AB - Intestinal atresia involving the ileocecal region is a very rare intestinal malformation, and the presence or absence of the ileocecal valve influences its surgical management. We report the case of a male newborn with a provisional diagnosis of distal ileal atresia, in whom laparotomy revealed that the entire ileocecal region was atretic with an absent ileocecal valve and appendix vermiformis. We resected the dilated terminal ileum together with the atretic segment and performed an ileocolic anastomosis between the terminal ileum and the transverse microcolon without valve reconstruction. When last seen, 8 months after the operation, the baby was developing normally. Ileocolic anastomosis without valve replacement appears to be sufficient if an ileocecal valve is completely absent and only a short segment of the terminal ileum is lost. PMID- 17123147 TI - Mucinous carcinoma of the duodenum associated with hereditary nonpolyposis colorectal cancer: report of a case. AB - We herein report a rare case of primary mucinous carcinoma of the duodenum associated with hereditary nonpolyposis colorectal cancer (HNPCC). A 50-year-old man known to have HNPCC based on the Amsterdam criteria I was admitted because of the presence of a duodenal tumor. Duodenoscopy revealed an ulcerated tumor in the posterior wall of the second portion of the duodenum and the malignancy was confirmed by a biopsy. He underwent a pylorus-preserving pancreaticoduodenectomy with a regional lymph node dissection. The histological diagnosis was mucinous carcinoma of the duodenum with lymph node metastasis. High-frequency microsatellite instability (MSI-H) was identified in both the colon and a duodenal specimen based on a microsatellite assay. A germline mutation in the hMSH2 gene was also identified. Even though extracolonic malignancies are associated with HNPCC, duodenal cancer is nevertheless very rare, and only two cases have been reported over the past 20 years. The present case is therefore only the third such case and the patient is herein described with a brief review of the literature. PMID- 17123148 TI - Complete response of highly advanced colon cancer with multiple lymph node metastases to irinotecan combined with UFT: report of a case. AB - Massive lymph node metastasis of the para-aortic region and supraclavicular lymph nodes, Virchow's lymph node metastasis due to colon cancer, is extremely rare. We herein report a case of such systemic lymph node metastasis that was successfully treated with a combination of irinotecan (CPT-11) and UFT, a combination drug of tegafur and uracil. The patient was a 57-year-old woman who had a tumor in the ascending colon, and massively swollen para-aortic and supraclavicular lymph node metastasis. She was treated with combination chemotherapy of CPT-11 and UFT. The main tumor was detected as a decompressed scar, and the supraclavicular and para aortic lymph nodes had completely disappeared after the second cycle of treatment. A histopathological examination and immunohistochemistry with cytokeratin showed complete remission of adenocarcinoma in the tumor and para aortic lymph nodes. She remains alive without recurrence 52 months after chemotherapy. Combination chemotherapy of CPT-11 and UFT may be of potential value in the treatment of advanced colorectal carcinoma, and both histopathological and immunohistochemical confirmation of a complete remission may indicate prolonged disease-free survival. PMID- 17123149 TI - John Money, Ph.D. (July 8, 1921-July 7, 2006): a personal obituary. PMID- 17123150 TI - Paraneoplastic neurological syndromes and breast cancer. Regression of paraneoplastic neurological sensorimotor neuropathy in a patient with metastatic breast cancer treated with capecitabine: a case study and mini-review of the literature. AB - Paraneoplastic neurological syndromes are a rare complication of breast cancer. Nevertheless, they may be clinically relevant leading to neurological impairment. Clinicians should be aware that these neurological disorders could even precede the diagnosis of breast cancer. Here we present the case of a female patient with advanced breast cancer who developed paraneoplastic sensorimotor neuropathy. Treatment with capecitabine lead to clinical amelioration. A review of the literature on the paraneoplastic neurological syndromes in breast cancer is also included. PMID- 17123151 TI - Weight gain and recovery of pre-cancer weight after breast cancer treatments: evidence from the women's healthy eating and living (WHEL) study. AB - PURPOSE: To examine predictors of weight gain following breast cancer diagnosis and subsequent return to pre-cancer weight. OBJECTIVES: To determine (1) the associations of anti-neoplastic chemotherapy and/or, Tamoxifen((R)) therapy on weight change following breast cancer diagnosis, (2) whether chemotherapy modified the effect of specific demographic and tumor characteristics on weight gain, (3) the proportion and characteristics of women who gained significant weight on chemotherapy and returned to their pre-cancer weight during follow-up. SUBJECTS AND METHODS: Participants were 3088 breast cancer survivors, aged 27-74 years. Weight was measured at baseline and years 1 through 6; pre-cancer weight was self-reported. Cancer stage and treatment modalities were obtained by medical record review; demographic and physical activity data were obtained from questionnaires. Weight gain of >/=5% body weight following cancer diagnosis was considered significant. RESULTS: Chemotherapy was significantly associated with weight gain (OR = 1.65, 95% CI = 1.12, 2.43) and Tamoxifen((R)) was not (OR = 1.03, 95% CI = 0.71, 1.51). Tamoxifen((R)) did not modify the effect of either chemotherapy or its different regimens on weight gain. Both types (anthracycline: OR = 1.63, p-value = 0.01, non-anthracycline: OR = 1.79, p = 0.003) and all regimens of chemotherapy (AC: OR = 1.55, p-value = 0.01, CAF: OR = 1.83, p = 0.003, CMF: OR = 1.76, p = 0.004) were associated with weight gain but the associations were not different from one another. Only 10% of participants returned to their pre-cancer diagnosis weight at the follow-up visits; the degree of initial gain (p for trend <0.0001) predicted that return. CONCLUSION: Chemotherapy was associated with clinically meaningful weight gain, and a return to initial weight following weight gain was unlikely. PMID- 17123152 TI - Differentially expressed genes between primary cancer and paired lymph node metastases predict clinical outcome of node-positive breast cancer patients. AB - The axillary lymph node status remains the most valuable prognostic factor for breast cancer patients. However, approximately 20-30% of node-positive patients remain free of distant metastases within 15-30 years. It is important to develop molecular markers that are able to predict for the risk of distant metastasis and to develop patient-tailored therapy strategies. We hypothesize that the lymph node metastases may represent the most metastatic fraction of the primary cancers. Therefore, we sought to identify the differentially expressed genes by microarray between the primary tumors and their paired lymph node metastases samples collected from 26 patients. A set of 79 differentially expressed genes between primary cancers and metastasis samples was identified to correctly separate most of primary cancers from lymph node metastases. And decreased expression of matrix metalloproteinase 2, fibronectin, osteoblast specific factor 2, collagen type XI alpha 1 in lymph node metastases were further confirmed by real-time RT-PCR performed on 30 specimen pairs. This set of genes also classified 35 primary cancers into two groups with different prognosis: "high risk group" and "low risk group." Patients in "high risk group" had a 4.65-fold hazard ratio (95% CI 1.02-21.13, P = 0.047) to develop a distant metastasis within 43 months comparing with the "low risk group." This suggested that the gene signature consisting of 79 differentially expressed genes between primary cancers and lymph node metastases could also predict clinical outcome of node positive patients, and that the molecular classification based on the gene signature could guide patient-tailored therapy. PMID- 17123153 TI - An unusual case of a ventral Richter's hernia at the site of a previous PEG tube. PMID- 17123154 TI - Sociodemographic effects on the dynamics of task-specific ADL functioning at the oldest-old ages: the case of China. AB - Studies that systematically examine the dynamics of task-specific ADL functioning and its associates are very rare. Using the first three waves of the Chinese Longitudinal Healthy Longevity Survey, this study examines the dynamics of each of the six ADL tasks (bathing, dressing, toileting, indoor transferring, eating, and continence) and their sociodemographic correlates among the oldest-old by including the ADL information both at the follow-up wave for survivors and at the time prior to death for those who died during survey intervals. Effects of age, gender, urban/rural residence, ethnicity, education, primary lifetime occupation, primary source of daily expenses, living alone, and marital status are examined in both the absence and presence of other various confounders. Our results show that each sociodemographic factor still plays some limited role in the dynamics of ADL functioning across tasks. PMID- 17123155 TI - Structural and functional properties of Cr 5, a new Lys49 phospholipase A2 homologue isolated from the venom of the snake Calloselasma rhodostoma. AB - Cr 5 PLA(2) homologous (K49) was isolated from Calloselasma rhodostoma venom in only one chromatographic step in reverse phase HPLC (RP-HPLC) (on mu-Bondapack C 18). A molecular mass of 13.965 Da was determined by MALDI-TOF mass spectrometry. The amino acid composition showed that Cr 5 had a high content of Lys, Tyr, Gly, Pro, and 14 half-Cys residues, typical residues of a basic PLA(2). The complete amino acid sequence of Cr 5 PLA(2) contains 120 residues, resulting in a calculated pI value of 5.55. This sequence shows high identity values when compared to other K49 PLA(2)s isolated from the venoms of viperid snakes. Lower identity is observed in comparison to D49 PLA(2)s. The sequence found was SLVELGKMIL QETGKNPAKS YGAYGCNCGV LGRHKPKDAT DRCCFVHKCC YKKLTGCDPK KDRYSYSWKD KTIVCGENNP CLKEMCECDK AVAICLRENL DTYNKKYRYL KPFCKKADDC. In mice, Cr 5 induced myonecrosis and edema upon intramuscular and intravenous injections, respectively. The LD(50) of Cr 5 was 0.070 mg/kg of the animal weight, by intracerebroventricular (i.c.v.) route. In vitro, the toxin caused rapid cytolytic effect upon mouse skeletal muscle myoblasts in culture. The isolation of this PLA(2) and the combined structural and functional information obtained classify Cr 5 as a new member of the K49 PLA(2) family, since it presents typical features from such proteins. PMID- 17123156 TI - The comparison question test: does it work and if so how? AB - In a mock crime study of the comparison question test (CQT), 35 subjects decided to participate as guilty and 30 as innocent. Two conditions were varied: Explaining the comparison questions in the pretest interview and re-discussing comparison questions between charts. Higher identification rates (approximately 90% for guilty and innocent participants) were achieved in groups with explanation of comparison questions than in groups without explanation. Re discussing comparison questions had no effect on identification rates. Ratings of subjective stress due to relevant and comparison questions were also obtained and can be seen as indicators of the significance of the questions. The significance of comparison questions was hardly affected by the different testing conditions. When effects are detectable at all, they contradict theoretical expectations in their direction. Results are discussed in terms of the significance of comparison questions used in polygraph testing. PMID- 17123157 TI - Psychopathy and violent crime: a prospective study of the influence of socioeconomic status and ethnicity. AB - The relationship between psychopathy and violence is well-established. However, few studies have examined the extent to which this relationship is influenced by sociodemographic predictors of violent criminality. In this prospective study we examine the power of psychopathy to predict criminal violence across ethnicity and levels of socioeconomic status in 199 European American and African American U.S. county jail inmates. A Psychopathy x SES x Ethnicity interaction was identified such that among European Americans psychopathy predicted recidivism at lower levels of SES but was unrelated to recidivism at higher levels of SES. The predictive power of psychopathy was relatively stable across SES among African Americans. The implications of our results for psychopathy and violence prediction are discussed. PMID- 17123158 TI - Race-based judgments, race-neutral justifications: experimental examination of peremptory use and the Batson challenge procedure. AB - Practically speaking, the peremptory challenge remained an inviolate jury selection tool in the United States until the Supreme Court's decision in Batson v. Kentucky. 476 U.S. 79 (1986). Batson's prohibition against race-based peremptories was based on two assumptions: (1) a prospective juror's race can bias jury selection judgments; (2) requiring attorneys to justify suspicious peremptories enables judges to determine whether a challenge is, indeed, race neutral. The present investigation examines these assumptions through an experimental design using three participant populations: college students, advanced law students, and practicing attorneys. Results demonstrate that race does influence peremptory use, but these judgments are typically justified in race-neutral terms that effectively mask the biasing effects of race. The psychological processes underlying these tendencies are discussed, as are practical implications for the legal system. PMID- 17123159 TI - The effect of lineup member similarity on recognition accuracy in simultaneous and sequential lineups. AB - Two experiments investigated whether remembering is affected by the similarity of the study face relative to the alternatives in a lineup. In simultaneous and sequential lineups, choice rates and false alarms were larger in low compared to high similarity lineups, indicating criterion placement was affected by lineup similarity structure (Experiment 1). In Experiment 2, foil choices and similarity ranking data for target present lineups were compared to responses made when the target was removed from the lineup (only the 5 foils were presented). The results indicated that although foils were selected more often in target-removed lineups in the simultaneous compared to the sequential condition, responses shifted from the target to one of the foils at equal rates across lineup procedures. PMID- 17123161 TI - Antioxidant activity of phenolics compounds from sugar cane (Saccharum officinarum L.) juice. AB - Phenolic compounds in sugar cane (Saccharum officinarum L.) juice were identified and quantified by analytical high performance liquid chromatography and photodiode array detection, showing the predominance of flavones (apigenin, luteolin and tricin derivatives), among flavonoids, and of hydroxycinnamic, caffeic and sinapic acids, among phenolic acids, representing a total content of around 160 mg/L. A tricin derivative was present in the highest proportion (>10% of the total). The phenolic extract obtained from sugar cane juice showed a protective effect against in vivo MeHgCl intoxication and potent inhibition of ex vivo lipoperoxidation of rat brain homogenates, indicating a potential use for beneficial health effects and/or therapeutic applications. PMID- 17123162 TI - Dietary onion intake as part of a typical high fat diet improves indices of cardiovascular health using the mixed sex pig model. AB - The aim of this study was to evaluate the potential health benefits of onions consumed at two levels of intake, using the pig model. The dietary fat content was set at a level typical of a "western" diet (25% w/w). Fifteen female and fifteen male pigs (Large White x Landrace) were allocated to one of three dietary treatments in a randomised block design. Treatments consisted of control diet (no onion) and onion supplementation at either 8.6 or 21.4 g of onion/MJ DE fed for six weeks. Onion consumption reduced plasma triglyceride levels by 15% (P=0.030) regardless of sex and onion dose. Total plasma cholesterol and cholesterol fractions were unaffected by onion supplementation (P>0.050). The bioactivity of onion was evident in haematocrit measures, where red blood cell and haemoglobin were significantly reduced in a dose dependant manner (P<0.001 and P=0.011, respectively), while other cell counts, with exception of segmented neutrophils ( 18%, P=0.012), were largely unaffected. Serum oxidative status was improved (P=0.007) in pigs consuming onions. These data demonstrate that consumption of onions can have positive health effects in both male and female pigs consuming a high fat diet. PMID- 17123163 TI - Amelioration of hexachlorocyclohexane-induced oxidative stress by amaranth leaves in rats. AB - The effect of prefeeding dehydrated amaranth leaves (AL), at 10 and 20% levels on hexachlorocyclohexane (HCH)-induced free radical stress in rat liver was evaluated. The HCH-induced raise in malonadialdehyde (MDA), conjugated dienes and hydroperoxides was diminished by AL. The effect of AL was highly effective with respect to reduction in these cytotoxic products, especially at 20% level. AL intake resulted in a significant increase in hepatic vitamin A and glutathione (GSH). However, the AL consumption reduced hepatic tocopherols. Feeding of AL at 10% level increased the hepatic glucose-6-phosphate dehydrogenase (G-6-PDH) activity while that at 20% level increased the hepatic glutathione reductase (GSSGR) as well, in addition to G-6-PDH. Amaranth leaves at 10 and 20% levels of feeding reduced the hepatic superoxide dismutase and glutathione peroxidase (GSH Px) activities. The pre-feeding of AL resulted in the reversal of HCH-induced alteration in GSH-Px and G-6-PDH activities. The significant reduction in the level of glutathione S-transferase brought about by HCH was restored to control level by feeding 20% AL. It is concluded that the consumption of AL at 20% level produces reduction in the HCH-induced impairment of antioxidant status in rat liver. PMID- 17123164 TI - Screening cell surface receptors using micromosaic immunoassays. AB - This report presents a general method for screening cell surface receptors using so-called micromosaic immunoassays. This method employs a microfluidic chip having n (n = 11) independent flow paths to move cells over m (m = 11) lines of surface-patterned antibodies for screening individual cells in a parallel, combinatorial, fast and flexible manner. The antibodies are patterned as 30 microm-wide lines on a poly(dimethylsiloxane) layer used to seal the area of the chip in which screening is being monitored. Mouse hybridoma cells having CD44 cell surface receptors and anti-CD44 antibodies were used to establish a proof-of concept for this method. Both the capture antibodies and the cells were fluorescently labelled to allow the position of the cells to be accurately tracked over the binding sites using an inverted fluorescence microscope. The chips and cells were maintained at a constant temperature between 20 to 37 degrees C, and flow velocities of the cells over the capture areas were 100-280 microm s(-1), resulting in a approximately 0.1-0.3 s residency time of the cells on each of the eleven 30 x 30 microm s2 capture areas. Binding of the cells appeared to be specific to the capture areas, with a yield of 30% when the assay was performed at a temperature of 37 degrees C and with a slow flow velocity. We suggest that this proof-of-concept is broadly applicable to the screening of cells for medical/diagnostic purposes as well as for basic research on the interaction of cells with surfaces. PMID- 17123165 TI - Chemoenzymatic synthesis of stable isotope labeled UDP-N-[2H]-acetyl-glucosamine and [2H]-acetyl-chitooligosaccharides. AB - Labeled UDP-GlcNAc and chitooligosaccharides should be powerful tools for studies of N-acetylglucosaminyltransferase such as chitin synthases. We describe here a rapid, inexpensive and a common strategie for the chemoenzymatic synthesis of uridine 5'-diphospho-N-[(2)H]-acetyl-glucosamine and the chemical preparation of N-[(2)H]-acetyl chitooligosaccharides (from 2 to 5 mers). Deuterated UDP-GlcNAc analogue was tested as chitin synthase substrate and it exhibited an incorporation level in chitin as the natural substrate. Deuterium labeling of carbohydrates present different advantages: it is a stable isotope and allows glycosyltransferase mechanism studies by a mass spectrometry approach. PMID- 17123166 TI - The human serotonin1A receptor exhibits G-protein-dependent cell surface dynamics. AB - Seven transmembrane domain G-protein-coupled receptors constitute the largest family of proteins in mammals. Signal transduction events mediated by such receptors are the primary means by which cells communicate with and respond to their external environment. The major paradigm in this signal transduction process is that stimulation of the receptor leads to the recruitment and activation of heterotrimeric GTP-binding proteins. These initial events, which are fundamental to all types of G-protein-coupled receptor signaling, occur at the plasma membrane via protein-protein interactions. As a result, the dynamics of the activated receptor on cell surfaces represents an important determinant in its encounter with G-proteins, and has significant impact on the overall efficiency of the signal transduction process. We have monitored the cell surface dynamics of the serotonin(1A) receptor, an important member of the G-protein coupled receptor superfamily, in relation to its interaction with G-proteins. Fluorescence recovery after photobleaching experiments carried out with the receptor tagged to the enhanced yellow fluorescent protein indicate that G protein activation alters the diffusion properties of the receptor in a manner suggesting the activation process leads to dissociation of G-proteins from the receptor. This result demonstrates that the cell surface dynamics of the serotonin(1A) receptor is modulated in a G-protein-dependent manner. Importantly, this result could provide the basis for a sensitive and powerful approach to assess receptor/G-protein interaction in an intact cellular environment. PMID- 17123167 TI - Alteration of the substrate specificity of Thermus caldophilus ADP-glucose pyrophosphorylase by random mutagenesis through error-prone polymerase chain reaction. AB - Expanding the scope of stereoselectivity is of current interest in enzyme catalysis. In this study, using error-prone polymerase chain reaction (PCR), a thermostable adenosine diphosphate (ADP)-glucose pyrophosphorylase (AGPase) from Thermus caldophilus GK-24 has been altered to improve its catalytic activity toward enatiomeric substrates including [glucose-1-phosphate (G-1-P) + uridine triphosphate (UTP)] and [N-acetylglucosamine-1-phosphate (GlcNAc) + UTP] to produce uridine diphosphate (UDP)-glucose and UDP-N-acetylglucosamine, respectively. To elucidate the amino acids responsible for catalytic activity, screening for UDP-glucose pyrophosphorylase (UGPase) and UDP-N-acetylglucosamine pyrophosphorylase (UNGPase) activities was carried out. Among 656 colonies, two colonies showed UGPase activities and three colonies for UNGPase activities. DNA sequence analyses and enzyme assays showed that two mutant clones (H145G) specifically have an UGPase activity, indicating that the changed glycine residue from histidine has the base specificity for UTP. Also, three double mutants (H145G/A325V) showed a UNGPase, and A325 was associated with sugar binding, conferring the specificity for the sugar substrates and V325 of the mutant appears to be indirectly involved in the binding of the N-acetylamine group of N acetylglucosmine-1-phosphate. PMID- 17123168 TI - Obituary: Hans Faillard (1924-2005). PMID- 17123169 TI - Isolation and characterization of acidic glycosphingolipids from the gill of the Pacific Salmon (Oncorhynchus keta): a novel hybrid-type ganglioside with isoglobo and neolacto-Series. AB - Monosialosyl gangliosides and sulfoglycolipids in the gill of pacific salmon, Oncorhynchus keta, have been prepared by solvent extraction and DEAE-Sephadex column chromatography. Acidic glycolipid bands (M1-M13) detected by thin layer chromatography were separated by Iatrobeads column chromatography and 13 components were characterized by TLC, compositional analysis, methylation analysis, chemical and enzymatic degradation, liquid secondary ion mass spectrometry and (1)H nuclear magnetic resonance spectroscopy. In addition to the acidic glycolipids with known structures (SM4s, SM3, GM3, LM1, GM1b and V(3)alphaFuc,IV(3)betaGalNAc-GM1a), two fractions (M11 and M13) of unknown monosialosyl gangliosides with TLC mobility slower than GM1a were isolated and characterized as having the following structure with a hybrid of isoglobo- and neolacto-series. [formula: see text] Analysis of fatty acid indicated predominance of C24:1 fatty acid in the upper band (M11) and shorter chain saturated fatty acids in the lower band (M13). The tissue concentrations of M11 and M13 were 1.15 and 0.96 mumol/kg wet weight, respectively. PMID- 17123170 TI - Is theory of mind understanding impaired in males with fragile X syndrome? AB - Males with fragile X syndrome (FXS) have difficulties with social interaction and many show autistic features. This study examined whether the social deficits characteristic of FXS are associated with theory of mind difficulties. Two groups of boys with FXS participated: a group with few autistic features and a group with many autistic features. An intellectual disability control group also participated. In addition to using standard theory of mind tasks, new techniques were used that were able to separate out the various processing demands of the task (e.g., memory, inhibitory control). Overall, the findings indicate that both groups of boys with FXS have difficulty with theory of mind tasks compared to an intellectual disability control group. However, both groups with FXS also performed worse on comparison trials that required working memory but not theory of mind. Theory of mind difficulties are likely to be an important aspect of the FXS clinical profile, but are most likely the result from a more basic difficulty with working memory. PMID- 17123171 TI - The role of indole and other shikimic acid derived maize volatiles in the attraction of two parasitic wasps. AB - After herbivore attack, plants release a plethora of different volatile organic compounds (VOCs), which results in odor blends that are attractive to predators and parasitoids of these herbivores. VOCs in the odor blends emitted by maize plants (Zea mays) infested by lepidopteran larvae are well characterized. They are derived from at least three different biochemical pathways, but the relative importance of each pathway for the production of VOCs that attract parasitic wasps is unknown. Here, we studied the importance of shikimic acid derived VOCs for the attraction of females of the parasitoids Cotesia marginiventris and Microplitis rufiventris. By incubating caterpillar-infested maize plants in glyphosate, an inhibitor of the 5-enolpyruvylshikimate-3-phospate (EPSP) synthase, we obtained induced odor blends with only minute amounts of shikimic acid derived VOCs. In olfactometer bioassays, the inhibited plants were as attractive to naive C. marginiventris females as control plants that released normal amounts of shikimic acid derived VOCs, whereas naive M. rufiventris females preferred inhibited plants to control plants. By adding back synthetic indole, the quantitatively most important shikimic acid derived VOC in induced maize odors, to inhibited plants, we showed that indole had no effect on the attraction of C. marginiventris and that M. rufiventris preferred blends without synthetic indole. Exposing C. marginiventris females either to odor blends of inhibited or control plants during oviposition experiences shifted their preference in subsequent olfactometer tests in favor of the experienced odor. Further learning experiments with synthetic indole showed that C. marginiventris can learn to respond to this compound, but that this does not affect its choices between natural induced blends with or without indole. We hypothesize that for naive wasps the attractiveness of an herbivore-induced odor blend is reduced due to masking by nonattractive compounds, and that during oviposition experiences in the presence of complex odor blends, parasitoids strongly associate some compounds, whereas others are largely ignored. PMID- 17123172 TI - (2S,12Z)-2-Acetoxy-12-heptadecene: major sex pheromone component of pistachio twig borer, Kermania pistaciella. AB - The sex pheromone of the pistachio twig borer, Kermania pistaciella (Lepidoptera: Oinophilidae), one of the most important insect pests of pistachio, Pistacia vera, in Turkey and Iran, was identified. In gas chromatographic electroantennographic detection (GC-EAD) and GC-mass spectrometric analyses of pheromone gland extracts of female K. pistaciella from Turkey, (2S,12Z)-2-acetoxy 12-heptadecene was identified as the major candidate pheromone component. In field experiments in Turkey, lures containing synthetic (2S,12Z)-2-acetoxy-12 heptadecene attracted large numbers of male moths. Its attractiveness was significantly reduced by the presence of the R-enantiomer or of either enantiomer of the corresponding alcohol. (2S,12Z)-2-Acetoxy-12-heptadecene is the first pheromone component identified in the Oinophilidae and the first secondary acetate pheromone component identified in the Lepidoptera. PMID- 17123173 TI - Medicine's moment of misrule: the medical student show. AB - Medical student shows are a prominent feature of medical student life around the world. Following a traditional vaudeville format of skits and songs, the shows are notorious for their exuberance, bawdiness, and lack of political correctness. Despite their widespread prevalence and sometimes hostile reactions, there has been no previous study of these shows. Based on research of scripts, programs, reviews, and oral history, this article explores their history and content and argues that, far from being irrelevant frivolities, these shows serve several important functions. These include the fostering of communal spirit, the development of skills in teamwork, and the collective ventilation of emotional reactions to the process of becoming a doctor. They are one offshoot of the ancient and important tradition of misrule in Western society. PMID- 17123174 TI - Sacred conceptions: clinical theodicies, uncertain science, and technologies of procreation in India. AB - This article argues that the rapid transfer of assisted conception technologies, such as in vitro fertilization, to India is not restricted merely to the modalities of offering potential biomedical resolution of infertility but includes, more crucially, how clinicians and infertile consumers assimilate the "Western technoscience" of conception. The article draws on a larger multisite ethnographic study of infertility and assisted conception in India's five major cities and is principally based on narratives of clinicians and infertile couples and on clinic-based ethnographic observations. In this article I contend that the success or failure of assisted conception, when situated in the universe of Hindu faith, becomes a powerful critique of the "incompleteness" of the "Western" science of conception. Situating this contention in the broader context of a clinician's faith, I assert that assisted conception--by conjoining seemingly disparate domains of the traditional and the modern, the sacred and the profane, the human and the superhuman, science and religion--produces clinical theodicies that help explain and contain the tentativeness permeating the conception technologies. The article concludes by arguing that this enchanted version of a thoroughly disenchanted worldview of biomedicine is part of a larger cultural process of indigenization of biomedicine in India. PMID- 17123175 TI - Molecular backgrounds of age-related osteoporosis from mouse genetics approaches. AB - Backgrounds underlying age-related bone loss can be classified into two categories: systemic abnormality and osteoblast dysfunction. The former includes insufficiency of vitamin D or estrogen, causing a negative balance of calcium metabolism. We propose the contribution of an aging-suppressing gene, klotho, as a novel systemic factor, as a mouse deficient in the klotho gene exhibits multiple aging phenotypes including osteopenia with a low bone turnover. As a factor intrinsic to osteoblasts, we investigated the role of PPARgamma, a key regulator of adipocyte differentiation, based on the facts that osteoblasts and adipocytes share a common progenitor. Heterozygous PPARgamma-deficient mice exhibited high bone mass by stimulating osteoblastogenesis from bone marrow progenitors, and this effect became prominent with aging, indicating involvement of PPARgamma-dependent bone formation in the pathophysiology of age-related bone loss. The local environment of osteoblasts is mainly controlled by cytokines/growth factors, among which insulin-like growth factor-I (IGF-I) is the most possible candidate whose production and activity are decreased with aging. Bone phenotypes of deficient mice of insulin receptor substrates (IRS-1 and IRS 2), essential molecules for intracellular signaling of IGF-I, revealed that IRS-1 is essential to maintain bone turnover by up-regulating anabolic and catabolic functions of osteoblasts, while IRS-2 is needed to keep the predominance of the anabolic function over the catabolic function. A next task ahead of us will be to elucidate the network system of these factors underlying age-related osteoporosis. PMID- 17123176 TI - Figurations of the real: representations of trauma in a dream. PMID- 17123177 TI - Reflections on Janet Zuckerman and Lisa Horelick's "the affective experience of the analyst in the extra-analytic moment". AB - A review and amplification of the article by authors Horelick and Zuckerman regarding the clinical phenomena of encountering patients (analysands) outside of the physical boundaries of the consultation room. Their clinical examples and the literature used to buttress their position and in particular the open disclosure of the deeply personal reactions they experienced are examined and used as launching points for further understanding. Ferenczi, Balint, Freud and other luminaries are incorporated into the topic of extra-analytic encounter, in an attempt to further demystify this inevitable and unplanned moment of disclosure, which, much like resistance and countertransference offer more opportunity than obstacle. PMID- 17123178 TI - Community-initiated urban development: an ecological intervention. AB - Neglected urban environments have been linked to social isolation, depression, and other health problems. In Portland, OR in 2003, an intervention was implemented and evaluated in three neighborhoods with the objective of promoting community participation in urban renewal and engaging residents in the construction of attractive urban places. Municipal officials approved and permitted community-designed street murals, public benches, planter boxes, information kiosks with bulletin boards, trellises for hanging gardens, all positioned in the public right-of-way. Residents within a two-block radius of the three sites were systematically sampled and interviewed before (N = 325) and after (N = 349) the intervention, of which, 265 individuals completed both surveys of the panel study. After the intervention, multivariate results revealed improvements in mental health (p = 0.03), increased sense of community (p < 0.01), and an overall expansion of social capital (p = 0.04). Through community empowerment, participation, and collective action, the strategy successfully engaged residents in restoring neighborhoods, with direct benefits to community well-being. PMID- 17123179 TI - Changing the location of care: management of patients with chronic conditions in Veterans Health Administration using care coordination/home telehealth. PMID- 17123180 TI - My vision. PMID- 17123181 TI - Location of plantar ulcerations in diabetic patients referred to a Department of Veterans Affairs podiatry clinic. AB - This study described the location of foot ulcerations via a retrospective chart review of diabetic patients in a Department of Veterans Affairs podiatry clinic and correlated location of ulceration with specific medical parameters. The heel was a site of ulceration in 11% of the patients. By multiple logistic regression, patients with diminished vascular function were more than five times more likely to have heel ulceration than patients with adequate vascular status. The findings suggest that heel ulcerations are more common than originally thought and are associated with diminished vascular status. Further work is necessary for reducing plantar heel pressure in individuals who are not presently candidates for vascular interventions. PMID- 17123182 TI - Effects of footwear on medial compartment knee osteoarthritis. AB - This pilot study investigated whether lateral-wedge insoles inserted into shock absorbing walking shoes altered joint pain, stiffness, and physical function in patients with symptomatic medial compartment knee osteoarthritis (OA). Twenty eight subjects wore full-length lateral-wedge insoles with an incline of 4 degrees in their walking shoes for 4 weeks. Pain, stiffness, and functional status were measured with the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index at baseline and 4 weeks postintervention. Significant improvements were observed in all three WOMAC subscales (pain, stiffness, and function). Pain scores were significantly reduced for the most challenging activity-stair climbing. Subjects wore insoles daily and tolerated them well. The results of this study indicated that lateral-wedge insoles inserted into shock absorbing walking shoes are an effective treatment for medial compartment knee OA. PMID- 17123183 TI - Isometric performance following total hip arthroplasty and rehabilitation. AB - We compared differences in isometric strength between older adults who have undergone elective unilateral total hip arthroplasty (THA) and completed rehabilitation with a population of community-dwelling older adults who have not had THA. The study was a cross-sectional design, and 22 unilateral THA subjects and 38 community-dwelling older adults participated. THA subjects received on average 13 outpatient or home-based physical therapy sessions before evaluation. THA subjects were evaluated 4 to 5 months postsurgery. We assessed isometric muscle strength by measuring peak hip torque per body weight with a robotic dynamometer during abduction, flexion, and extension. No significant performance differences were observed between operated and nonoperated hips of THA subjects. THA subject operated and nonoperated hips generated significantly less peak torque per body weight during flexion (p = 0.03) compared with community-dwelling older adult hips (THA subject operated hips = 6.96 ft-lb/lb, THA subject nonoperated hips = 8.26 ft-lb/lb, community-dwelling older adult hips = 11.56 ft lb/lb). No significant differences were observed between THA subjects and community-dwelling older adults during hip extension (p = 0.55) or abduction (p = 0.17). At 4 to 5 months postsurgery, THA subjects were not at the same level of biomechanical performance as community-dwelling older adults. Significant strength deficits were found in THA subject operated versus nonoperated hips during isometric flexion. Additional or modified physical therapy that targets the hip flexors is recommended after THA. PMID- 17123184 TI - Influence of a 6-week arm exercise program on walking ability and health status after hip arthroplasty: a 1-year follow-up pilot study. AB - The influence of an upper-limb interval-training program after total hip arthroplasty (THA) in elderly patients was studied during a 1 yr follow-up on health status and walking ability. After surgery, 14 patients were randomly assigned to the control group that started a 6 wk general rehabilitation program or the training group that combined it with an interval exercise program on an arm ergometer. A Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index was completed 1 mo before and 2 mo and 1 yr after surgery. A 6 min walking test was performed at 2 mo and 1 yr after surgery. The training group covered a significantly longer distance than the control group in the 6 min walk test and obtained significantly lower WOMAC scores both at 2 mo and 1 yr after surgery. However, both groups significantly improved in WOMAC and in their performance during the survey. Endurance-type upper-body aerobic training in a rehabilitation program might be important after THA. PMID- 17123185 TI - Are patient ratings of chronic pain services related to treatment outcome? AB - Consumer ratings of satisfaction with treatment are rarely used as measures of treatment outcome. This study examined the relationships between service ratings and psychometric outcomes of patients receiving pain-management services in a tertiary teaching hospital. A group of 122 patients who completed a multidisciplinary pain-management program rated their satisfaction with and effectiveness of services received and changes in their pain condition and quality of life (QOL). They also completed pre- and posttreatment measures of pain severity, pain interference, depression, and disability. Pain severity, pain interference, and depression significantly decreased following treatment. The patients' ratings of services were significantly associated with outcome measures. Pre- to posttreatment changes in pain severity and pain interference were associated with treatment satisfaction and effectiveness, improvement in pain condition, and QOL. Pre- to posttreatment change in disability was significantly related to ratings of treatment effectiveness, improvement in pain condition, and quality of life. The findings suggest that pain intensity, pain interference, and disability are important outcome dimensions of pain-management programs. PMID- 17123186 TI - Using cranial electrotherapy stimulation to treat pain associated with spinal cord injury. AB - Treatments for chronic pain in persons with spinal cord injury (SCI) have been less than effective. Cranial electrotherapy stimulation (CES), a noninvasive technique that delivers a microcurrent to the brain via ear clip electrodes, has been shown to effectively treat several neurological and psychiatric disorders. The present study examined the effects of daily 1-hour active CES or sham CES treatment (randomly assigned) for 21 days on pain intensity and interference with activities in 38 males with SCI. The active CES group (n = 18) reported significantly decreased daily pain intensity compared with the sham CES group (n = 20) (mean change: active CES = -0.73, sham CES = -0.08; p = 0.03). Additionally, the active CES group reported significantly decreased pain interference (-14.6 pre- vs postintervention, p = 0.004) in contrast to the nonsignificant decrease in the sham CES group (-4.7 pre- vs postintervention, p = 0.24). These results suggest that CES can effectively treat chronic pain in persons with SCI. PMID- 17123187 TI - Race/ethnicity: who is counting what? AB - Misclassification of race and ethnicity in administrative data may produce misleading results if it is overlooked or ignored. In this study, we examined the racial/ethnic classifications of 1,084 veterans with stroke in Florida who received inpatient and outpatient services within the Department of Veterans Affairs (VA) healthcare system and who were also eligible for Medicare between 2000 and 2001. We compared the reliability of racial/ethnic classifications between VA inpatient data, VA outpatient data, and Medicare data. Our results showed that (1) the rate of unknown racial/ethnic classification in VA outpatient and inpatient data was high, (2) minimizing the unknowns by substituting known values from other data when available would greatly enhance the overall and individual classification reliability, (3) black and white classifications in the VA data had stronger agreement with Medicare data, and (4) Medicare data may under-represent Hispanic patients. PMID- 17123188 TI - Treadmill training with harness support: selection of parameters for individuals with poststroke hemiparesis. AB - Locomotor training with a treadmill and harness support is a promising, task oriented approach to restoring gait function in individuals with poststroke hemiparesis. However, a scientific basis for the proper selection of training parameters is lacking. Considerable latitude exists in the application of locomotor training, and training protocols vary widely between experimenters and clinical settings. Recent studies indicate that the prescription of certain parameters, including body-weight support (BWS) and treadmill speed, can affect treatment outcome in hemiparetic individuals. As an initial step toward developing a basis for selection of parameters, we reviewed the literature for studies that quantified the immediate (i.e., within session) biomechanical effects of adjusting BWS, treadmill speed, support stiffness, and handrail hold during treadmill walking in hemiparetic and nondisabled subjects. We then summarized results from personal investigations of these parameters. Based on the currently available evidence, we discuss the scientific rationale for selecting certain training parameters for individuals with poststroke hemiparesis and outline future directions for research. PMID- 17123189 TI - Caregiver distress in parkinsonism. AB - This study examined the frequency and degree of caregiver burden in persons with parkinsonism, a group of disorders with four primary symptoms that include tremor, rigidity, postural instability, and bradykinesia. We assessed associations between perceived caregiver burden and physical, cognitive, and functional impairments using well-established tools for persons with parkinsonism. The 49 individuals with parkinsonism ranged in age from 61 to 87 (mean = 75), while their caregivers (N = 49) ranged in age from 48 to 83 (mean = 70). The caregivers were predominantly either wives (82%) or daughters (6%), with other family members, friends, and/or neighbors (12%) making up the rest. The caregivers reported a relatively high ability for coping (mean scores = 4.6/6). Caregiver burden was significantly negatively associated with activities of daily living and motoric difficulties as measured on the Unified Parkinson's Disease Rating Scale (UPDRS). Likewise, caregiver burden was negatively associated with caregiver self-reported sleep and coping ability. Results did not demonstrate an association on the UPDRS among mentation, behavior, and mood. We found a significant negative correlation for mentation between the Folstein Mini-Mental Status Examination and caregiver burden measures; however, we did not find this association with the Dementia Rating Scale-2. Patient's self-reported pain and caregiver burden were not associated. PMID- 17123190 TI - A noncontact sensor for measurement of distal residual-limb position during walking. AB - A simple noncontact device was implemented for measuring the position of the distal residual limb relative to the prosthetic socket during ambulation. The device was a small and lightweight photoelectric sensor positioned within a frame mounted immediately beneath the socket. Calibration tests showed that the sensor had a displacement range of 60.0 mm. The root-mean-square error for all sources of error considered (different reflective surfaces, peak-to-peak signal noise, drift, nonlinearity, different surface tilt angles, surface curvature, and wetness [simulating sweating]) was <1.95% full-scale output. We used the sensor in a preliminary study on a unilateral, transtibial amputee with diabetes to assess pistoning during ambulation. Results showed an average 41.7 mm proximal displacement during swing phase relative to stance phase. When the subject was walking on a flat surface, pistoning was significantly less (p = 0.000) with a supracondylar strap compared with no supracondylar strap, although the difference was not substantial (0.8 mm). A 5 min rest period caused the limb to displace proximally in the socket approximately 4.8 mm during subsequent walking trials, possibly reflecting limb enlargement and thus a more proximal position in the socket after the rest period. The device can potentially be used in prosthetics research for evaluating clinical features that may affect limb position and pistoning and thus fit. PMID- 17123191 TI - Acclimatization in wide dynamic range multichannel compression and linear amplification hearing aids. AB - Acclimatization was studied in hearing-impaired patients with no previous hearing aid (HA) experience who were fit bilaterally with either wide dynamic range multichannel compression (WDRMCC) or linear amplification (LA) HAs. Throughout 40 weeks of normal HA use, we monitored changes in nonsense syllable perception in speech-spectrum noise. Syllable recognition for WDRMCC users improved by 4.6% over the first 8 weeks, but the 2.2% improvement for LA users was complete in 2 to 4 weeks. Consonant confusion analyses indicated that WDRMCC experience facilitated consonant identification, while LA users primarily changed their response biases. Furthermore, WDRMCC users showed greater improvement for aided than unaided stimuli, while LA users did not. These results demonstrate acclimatization in new users of WDRMCC HAs but not in new users of LA HAs. A switch in amplification type after 32 weeks produced minimal performance change. Thus, acclimatization depended on the type of amplification and the previous amplification experience. PMID- 17123192 TI - Perceptual training improves syllable identification in new and experienced hearing aid users. AB - We assessed the effects of perceptual training of syllable identification in noise on nonsense syllable test (NST) performance of new (Experiment 1) and experienced (Experiment 2) hearing aid (HA) users with sensorineural hearing loss. In Experiment 1, new HA users were randomly assigned to either immediate training (IT) or delayed training (DT) groups. IT subjects underwent 8 weeks of at-home syllable identification training and in-laboratory testing, whereas DT subjects underwent identical in-laboratory testing without training. Training produced large improvements in syllable identification in IT subjects, whereas spontaneous improvement was minimal in DT subjects. DT subjects then underwent training and showed performance improvements comparable with those of the IT group. Training-related improvement in NST scores significantly exceeded improvements due to amplification. In Experiment 2, experienced HA users received identical training and testing procedures as users in Experiment 1. The experienced users also showed significant training benefit. Training-related improvements generalized to untrained voices and were maintained on retention tests. Perceptual training appears to be a promising tool for improving speech perception in new and experienced HA users. PMID- 17123193 TI - Interface pressure and cutaneous hemoglobin and oxygenation changes under ischial tuberosities during sacral nerve root stimulation in spinal cord injury. AB - Noninvasive functional magnetic stimulation (FMS) of the sacral nerve roots can activate gluteal muscles. We propose the use of sacral anterior root stimulator (SARS) implants to prevent ischial pressure ulcers in the spinal cord injury (SCI) population. In this study, we (1) investigated the acute effects of sacral FMS on ischial pressure, skin blood content, and oxygenation changes in people with SCI and demonstrated the utility of FMS as an assessment tool, and (2) showed that similar effects are possible with electrical stimulation via a SARS implant. Results indicated that sacral nerve root stimulation, either by FMS or implanted electrical stimulation, induced sufficient gluteus maximus contraction to significantly change subjects' ischial pressures and cutaneous hemoglobin and oxygenation during sitting. In addition to these beneficial acute effects, chronic stimulation via a SARS implant may build gluteal muscle bulk and prevent or reduce pressure ulcers in the SCI population. PMID- 17123194 TI - Antinociceptive effect of linear polarized 0.6 to 1.6 microm irradiation of lumbar sympathetic ganglia in chronic constriction injury rats. AB - Linear polarized near-infrared light created with linear polarized near-infrared light therapy equipment (Super Lizer HA-550, Tokyo Iken Co, Ltd, Tokyo, Japan) has been used for the treatment of various painful disorders in Japan. Irradiation near the stellate ganglion with a Super Lizer (ISGL) is an especially notable therapeutic method used with stellate ganglion block (SGB) or substitutes for SGB. ISGL is a safe, simple, well-tolerated, and effective treatment. We examined the effects of irradiation with a Super Lizer applied to an area near the lumbar sympathetic ganglia on the ligated side in a chronic constriction injury (CCI) model, which is believed to be an animal model of complex regional pain syndrome (CRPS). Rats showing thermal hyperalgesia in a radiant heat test 1 wk postoperatively were used in Experiments 1 and 2: (1) Thermal hyperalgesia of irradiation group (n = 11) was less than that of the control or nonirradiation (n = 11) group at 1, 3, and 8 h after irradiation; however, the effect disappeared 12 h after irradiation. (2) Daily irradiation (n = 16) and 1 wk (n = 14) from 7 days after nerve ligation significantly shortened the interval from thermal hyperalgesia until recovery. Rats showing mechanical hyperalgesia in the von Frey hair test 1 wk postoperatively were used in Experiment 3: 1 wk irradiation beginning 7 days after nerve ligation (n = 9) did not promote the recovery from mechanical hyperalgesia. We speculate that repeated ISGL may be more effective than a single ISGL in alleviating pain in CRPS patients. We cannot explain the discrepancy between the results obtained in Experiments 2 and 3. We believe the results of this study are relevant to the effect of ISGL for patients with upper limb CRPS: irradiation near the lumbar sympathetic ganglia of the rat is effective for thermal but not mechanical pain in CCI. PMID- 17123195 TI - Skin and bone integrated prosthetic pylon: a pilot animal study. AB - Direct skeletal attachment of limb prostheses is a viable alternative to traditional techniques that are based on a socket-residuum interface. Direct skeletal attachment may be a better or even the only method for patients with a very short residuum and high soft-tissue volume. The problem of integrating the prosthetic pylon with residual skin during direct skeletal attachment of a limb prosthesis has not been solved, and the use of a completely porous prosthetic pylon has not been the subject of focused, systematic research. In this in vivo study, we investigated cell (osteocyte, fibroblast, and keratinocyte) adhesion and penetration into the pores of a titanium pylon implanted in Wistar rats. The porous titanium pylon was implanted in the bone of the thigh residua of four rats. Electronic scanning and morphological analysis demonstrated integration of the pylon with the surrounding skin. These findings support the possibility of developing a natural barrier against the infection associated with direct skeletal attachment of limb prostheses. PMID- 17123196 TI - Those magnificent men and their flying machines. PMID- 17123197 TI - Robotics and clinical research: Collaborating to expand the evidence base for rehabilitation. PMID- 17123198 TI - Rehabilitation robotics research at the National Institute on Disability and Rehabilitation Research. PMID- 17123199 TI - National Institutes of Health support of rehabilitation robotics research. PMID- 17123200 TI - Passive exoskeletons for assisting limb movement. AB - This article presents the state of the art in passive devices for enhancing limb movement in people with neuromuscular disabilities. Both upper- and lower-limb projects and devices are described. Special emphasis is placed on a passive functional upper-limb orthosis called the Wilmington Robotic Exoskeleton (WREX). The development and testing of the WREX with children with limited arm strength are described. The exoskeleton has two links and 4 degrees of freedom. It uses linear elastic elements that balance the effects of gravity in three dimensions. The experiences of five children with arthrogryposis who used the WREX are described. PMID- 17123201 TI - Principle and design of a mobile arm support for people with muscular weakness. AB - This article describes the development of a mobile arm support for people with muscular diseases. The arm support is spring-balanced, with special attention on reduction of operating effort (high balancing quality and low friction), functionality (large range of motion), and aesthetics (inconspicuous design). The spring settings can be adjusted for wearing heavier clothing or picking up an object, a function that can also be used for moving up or down. The device levels itself automatically to compensate for uneven floors, a function that can be overruled to assist forward/backward motion of the arm. Thus, the balancer can compensate for the weight of the arm and be adjusted to generate force to a limited (safe) extent. The principle and design of the mechanism are presented and preliminary field trial results are given. Two users report on 6 months of continuous use of the arm support in their home and social environments. PMID- 17123202 TI - Motions or muscles? Some behavioral factors underlying robotic assistance of motor recovery. AB - Robotics and related technologies have begun to realize their promise to improve the delivery of rehabilitation therapy. However, the mechanism by which they enhance recovery remains unclear. Ultimately, recovery depends on biology, yet the details of the recovery process remain largely unknown; a deeper understanding is important to accelerate refinements of robotic therapy or suggest new approaches. Fortunately, robots provide an excellent instrument platform from which to study recovery at the behavioral level. This article reviews some initial insights about the process of upper-limb behavioral recovery that have emerged from our work. Evidence to date suggests that the form of therapy may be more important than its intensity: muscle strengthening offers no advantage over movement training. Passive movement is insufficient; active participation is required. Progressive training based on measures of movement coordination yields substantially improved outcomes. Together these results indicate that movement coordination rather than muscle activation may be the most appropriate focus for robotic therapy. PMID- 17123203 TI - Robot-assisted movement training for the stroke-impaired arm: Does it matter what the robot does? AB - Robot-assisted movement training improves arm movement ability following acute and chronic stroke. Such training involves two interacting processes: the patient trying to move and the robot applying forces to the patient's arm. A fundamental principle of motor learning is that movement practice improves motor function; the role of applied robotic forces in improving motor function is still unclear. This article reviews our work addressing this question. Our pilot study using the Assisted Rehabilitation and Measurement (ARM) Guide, a linear robotic trainer, found that mechanically assisted reaching improved motor recovery similar to unassisted reaching practice. This finding is inconclusive because of the small sample size (n = 19), but suggest that future studies should carefully control the amount of voluntary movement practice delivered to justify the use of robotic forces. We are optimistic that robotic forces will ultimately show additional therapeutic benefits when coupled with movement practice. We justify this optimism here by comparing results from the ARM Guide and the Mirror Image Movement Enabler robotic trainer. This comparison suggests that requiring a patient to generate specific patterns of force before allowing movement is more effective than mechanically completing movements for the patient. We describe the engineering implementation of this "guided-force training" algorithm. PMID- 17123204 TI - MIME robotic device for upper-limb neurorehabilitation in subacute stroke subjects: A follow-up study. AB - This study presents results from a randomized controlled clinical trial of the Mirror Image Movement Enabler (MIME) robotic device for shoulder and elbow neurorehabilitation in subacute stroke patients, including data on the use of its bilateral training mode. MIME incorporates a PUMA 560 robot (Staubli Unimation Inc, Duncan, South Carolina) that applies forces to the paretic limb during unilateral and bilateral movements in three dimensions. Robot-assisted treatment (bilateral, unilateral, and combined bilateral and unilateral) was compared with conventional therapy. Similar to a previous study in chronic stroke, combined unilateral and bilateral robotic training had advantages compared with conventional therapy, producing larger improvements on a motor impairment scale and a measure of abnormal synergies. However, gains in all treatment groups were equivalent at the 6-month follow-up. Combined unilateral and bilateral training yielded functional gains that were similar to the gains from equivalent doses of unilateral-only robotic training, although the combined group had more hypertonia and less movement out of synergy at baseline. Robot-assisted treatment gains exceeded those expected from spontaneous recovery. These results are discussed in light of the need for further device development and continued clinical trials. PMID- 17123205 TI - Custom-designed haptic training for restoring reaching ability to individuals with poststroke hemiparesis. AB - We present an initial test of a technique for retraining reaching skills in patients with poststroke hemiparesis, in which errors are temporarily magnified to encourage learning and compensation. Individuals with poststroke hemiparesis held a horizontal plane robotic manipulandum that could exert a variety of forces while recording patients' movements. We measured how well the patients recovered movement straightness in a single visit to the laboratory (approximately 3 h). Following training, we returned forces to zero for an additional 50 movements to discern if aftereffects lasted. We found that all subjects showed immediate benefit from the training, although 3 of the 10 subjects did not retain these benefits for the remainder of the experiment. We discuss how these approaches demonstrate great potential for rehabilitation tools that augment error to facilitate functional recovery. PMID- 17123206 TI - Tools for understanding and optimizing robotic gait training. AB - This article reviews several tools we have developed to improve the understanding of locomotor training following spinal cord injury (SCI), with a view toward implementing locomotor training with robotic devices. We have developed (1) a small-scale robotic device that allows testing of locomotor training techniques in rodent models, (2) an instrumentation system that measures the forces and motions used by experienced human therapists as they manually assist leg movement during locomotor training, (3) a powerful, lightweight leg robot that allows investigation of motor adaptation during stepping in response to force-field perturbations, and (4) computational models for locomotor training. Results from the initial use of these tools suggest that an optimal gait-training robot will minimize disruptive sensory input, facilitate appropriate sensory input and gait mechanics, and intelligently grade and time its assistance. Currently, we are developing a pneumatic robot designed to meet these specifications as it assists leg and pelvic motion of people with SCI. PMID- 17123207 TI - Machines to support motor rehabilitation after stroke: 10 years of experience in Berlin. AB - The group at Klinik Berlin/Charite University Hospital in Berlin, Germany, began studies to promote motor recovery after stroke in the early 1990s. Following the introduction of treadmill training with partial body-weight support, the group designed an electromechanical gait trainer, GT I, based on movable foot plates that relieve therapist effort (e.g., when assisting the movement of the paretic limbs) and intensify patient gait training (GT). Preliminary results of a recent multicenter trial of 155 acute stroke patients showed that the GT I effectively promotes gait ability and competence in activi ties of daily living. The experimental group received 20 min of GT and 25 min of physiotherapy (PT) and the control group received 45 min of PT every day for 4 weeks. The laboratory's next step was the HapticWalker, a robotic walking simulator with freely programmable foot plates so that patients can, for example, additionally train for stair climbing and perturbations. The foot plates can be operated in full guidance or compliance control modes, thus reducing foot support according to the patient's learning success. For the severely affected upper limb, the laboratory's computerized arm trainer, called the Bi-Manu-Track, enables bilateral practice of forearm pronation/supination and wrist flexion/extension. Compared with electrical stimulation of the paretic wrist extensors, acute stroke patients with severe arm paresis (n = 44) had significantly more upper-limb muscle strength and control at the end of the 6-week intervention period and at follow-up. The laboratory's most recent and cost-effective development, the Nudelholz, is a purely mechanical device that bilaterally trains the shoulder, elbow, and wrist joints. It is intended for home therapy. PMID- 17123208 TI - Human-centered robotics applied to gait training and assessment. AB - Robot-aided gait training can increase the duration and number of training sessions while reducing the number of therapists required for each patient. However, current automated gait trainers do not adapt their movement to the patient's muscular efforts and passive musculoskeletal properties. Furthermore, robot-aided training without therapists lacks the feedback required for patient assessment. In this article, we present results from the literature and our research to provide an overview of novel human-centered strategies for robot behaviors that are patient-cooperative and support motor-function assessment. Combining robot-aided training with robot-aided assessment will likely make future gait therapy easier, more comfortable, and more efficient. Broad clinical testing is still required for proving this assumption. PMID- 17123209 TI - Telerehabilitation robotics: bright lights, big future? AB - The potential for remote diagnosis and treatment over the Internet using robotics is now a reality. The state of the art is exemplified by several Internet applications, and we explore the current trends in developing new systems. We review the technical challenges that lie ahead, along with some potential solutions. Some promising results for a new bilateral system involving two InMotion2 robots are presented. Finally, we discuss the future direction and commercial outlook for rehabilitation robots over the next 15 years. PMID- 17123210 TI - Gastric ulcer penetrating into the heart. PMID- 17123211 TI - Perforation of the esophagus after dilation treatment for dysphagia in a patient with eosinophilic esophagitis. PMID- 17123212 TI - Resection of esophageal leiomyoma using an endoscopic submucosal dissection technique. PMID- 17123214 TI - [Long-term efficiency of orthopedic rehabilitation in chronic back pain--the integrative orthopedic psychosomatic concept (IopKo)]. AB - Chronic diseases of the musculoskeletal system rank first as causes of early retirement in Germany. Therefore orthopaedic rehabilitation has to identify patients with work-related problems and to promote return to work through differential treatment and vocational counselling. In the framework of the IopKo Project such measures were developed and evaluated. These measures encompass: (1) an intensive and multiprofessional diagnostic pathway which allows early detection and treatment of mental disorders and job related problems; (2) homogeneous treatment groups based on multiprofessional diagnostics; (3) differential treatments, among these a multidisciplinary programme for patients with chronic low back pain or high risk of chronification (Ruckenfit); (4) interactive training modules which mediate principles of performance and disability expertise, the legal bases of retirement pensioning, and measures to support occupational rehabilitation; and (5) a work hardening training programme. METHOD: To evaluate the effects of these measures in comparison to a control group with usual care, a prospective longitudinal study was conducted. A total of 307 patients were assigned to the multidisciplinary in-patient treatment programme, whereas 176 patients in the control group had a standard rehabilitation programme. RESULTS: The results show positive moderate and strong effects in the intervention group concerning function, pain, psychic strains as well as the number of sick days and return to work rates 10 months after discharge. The effects in the intervention group exceeded the effects achieved in the control group. PATIENTS WITH HIGH RISK OF CHRONIFICATION: Beside the full sample, a subgroup of patients with chronic pain or high risk of chronification was analyzed, who had received a multidisciplinary functional restoration treatment. Also for this subgroup we found moderate and strong effects of treatment for function, psychic strains and sick days superior to those in the control group. CONCLUSION: By this study we were able to show that orthopedic rehabilitation in a multimodal and multidisciplinary setting with a focus on activating and motivating therapy can have sustainable positive effects on pain, function and psychic well-being as well as on economic parameters. We interpret these persistent and superior effects in the treatment group (1) as a result of multiprofessional diagnosis and assignment which helps to subdivide the inhomogeneous group of patients with unspecific back pain into more homogeneous and thus more effective subgroups, (2) as a result of increased motivation by closed treatment groups, (3) as a result of intense and multilevel counselling of work related problems, (4) as a result of work hardening modules, and (5) as a result of direct and efficient treatment of psychic strains. The results also demonstrate the significance of inpatient rehabilitation, which will be efficient if differential treatment - adequate to the problems of the patient - is offered. PMID- 17123215 TI - [Chronic low back pain with endurant pain coping--optimizing rehabilitation success in a hitherto neglected patient group]. AB - In view of a frequently poor success rate of rehabilitation programmes in patients with low back pain, a specific intervention programme was developed addressing the subgroup of patients with endurant pain coping behaviour and evaluated within a research project sponsored by the National federation of pension insurers, VDR. In terms of a control group design with 4 points of measurement (beginning and end of rehabilitation, 6- and 12-months follow-up), an intervention group (n = 163) was compared to a control group (standard programme of an orthopaedic rehabilitation clinic, n = 252). Data were collected using the KSI, BDI, IRES, FFbH-R and a pain questionnaire. Subjects were blue-collar insurants, aged 46 on average with a pain history of 9.6 years. Findings show a stronger reduction of pain intensity and strain, significant enhancement of functional capacity as well as a greater decrease of sick leave days and pension requests. These results are presented over the course of the 4 points of measurement as well as in relation to the intervention programme, which focuses on the endurant pain coping behaviour. The consequences of implementing such therapy components into standard rehabilitation programmes are discussed. PMID- 17123216 TI - [On uniform social medical evaluation of performance capacity in orthopedic rehabilitation]. AB - The need for standardized and unified performance capacity assessment in orthopedic rehabilitation has led to the attempt to use an existing standardized procedure. Graduated disease features have been defined which are relevant for work and their effects on performance capacity been formulated as rules. In this way so-called disease-conditioned performance capacities are obtained which help to identify problems in carrying out various activities. These activities can be directly assessed using tests. Appropriate tests are listed in a table. PMID- 17123217 TI - [Retrospective analysis of falls in an inpatient rehabilitation centre for rheumatic and orthopaedic diseases]. AB - AIM: To analyse the frequency and circumstances of falls in an inpatient rehabilitation centre. METHODS: In a survey all falls reported to the Austrian accident insurance carrier were analysed and external circumstances of falls were evaluated. The survey covers a period of 64 months (Jan 2000 to April 2005): 10 820 inpatients (234 502 patient days); mean age 52.8 years; 1,252 patients aged >/= 70 years; 47 % male, 53 % female. RESULTS: Of 223 accidents registered, 140 (62.8 %) were falls. Of these, 17 patients had to be referred to a surgical unit for further treatment. In total, 1.29 % of all patients fell during their stay (with report to the accident insurance carrier), 0.16 % suffered major injuries from falls (with referral to surgical unit). Females fell more frequently than males. 39 % of the falls were associated with physical therapy. Most falls occurred in daytime. The location in the centre or the day of week showed no major influence on the frequency of falls. Falls occurred slightly more often in the first part of the three-week inpatient stay. CONCLUSION: The high total number of falls shows that falls are important in an inpatient rehabilitation centre for rheumatic and orthopaedic diseases. The analysis did not reveal any special location inside the rehabilitation centre with an increased risk of falls calling for urgent interventions. The active and activating physical therapies do not seem to increase the number of falls significantly. PMID- 17123218 TI - [Pre- and post-treatment in inpatient psychosomatic rehabilitation]. AB - Patients who are referred to psychosomatic inpatient rehabilitation are mostly suffering from long-term illnesses. Therefore, it is necessary to take pre- and post-treatment into account. Epidemiological questions are frequency and type of pre- and post-treatment and their relation to course and outcome of rehabilitation. Pre-treatment, recommendations for follow-up treatment, and course of treatment in 1284 patients of the department of behaviour therapy and psychosomatics of the rehabilitation centre Seehof were assessed. Before admission, 75.4 % of patients had been in psychiatric treatment, and 31.3 % had received psychotherapy. The inpatient stay was significantly longer in patients with psychiatric pre-treatment and those with a combination of psychiatric treatment and psychotherapy. These patients were more often unemployed and showing an insufficient social network. Pre-treatment was unrelated to changes in the SCL-90-R, to physician ratings of therapeutic outcome or the ability to work at the end of rehabilitation. Patients pre-treated by a general practitioner, psychiatrist or psychotherapist returned to that therapeutic setting after rehabilitation. Patients only treated by general practitioners were referred to specialist treatment. These data show the need to integrate inpatient rehabilitation in long-term pre- and post-treatment. One effect of inpatient rehabilitation is to optimize treatment. This requires good communication across different areas of health care. PMID- 17123219 TI - [Effects of therapy goal oriented cognitive behavioural aftercare measures on therapy transfer following inpatient psychosomatic rehabilitation]. AB - OBJECTIVE: After discharge from inpatient psychosomatic rehabilitation patients often face problems to transfer and stabilize the modifications they have achieved in the clinic. Using targeted outpatient aftercare interventions up to eight weeks after discharge from the inpatient programme we tried to support this transfer, thus enhancing treatment effectiveness. METHOD: The evaluation was carried out as a field experiment. Patients were matched into pairs and then allocated randomly to either a control or an experimental condition. The experimental group, which had participated in specific aftercare measures after inpatient therapy, was compared to the control group at "discharge from clinic" and "twelve weeks post-discharge" relative to a number of variables relevant in therapy outcome. RESULTS: Twelve weeks post-discharge patients who had participated in aftercare measures show significantly better progress relative to their physical and psychological complaints than patients in the control group. Also, they are more capable of coping with psychosocial demands of their everyday life. They experience better control of their symptoms and they are more successful in attaining the goals they have set for themselves for the time after discharge. CONCLUSIONS: The findings show that supplementing psychosomatic inpatient rehabilitation with specific aftercare measures will lead to better transfer and to increased effectiveness of psychosomatic rehabilitation. Also on account of our results, outpatient rehab aftercare programmes have in the meantime become a regular component of inpatient psychosomatic rehabilitation in Germany. PMID- 17123220 TI - [Aspects of sample size determination and power calculation illustrated on examples from rehabilitation research]. AB - Often it is reported in medical studies that an expected effect could not be detected. This may be the case if the sample size had been too small to detect an effect which actually exists. This often is due to the fact that sound sample size estimation had been omitted prior to the study outset. As a result, it is not known how many persons should have been involved in the study to detect this effect if present. On the other hand, if sample size estimation has not been realized, more persons than needed might be included in the study. This is problematic for economic and in particular for ethical reasons. The aim of this paper is to point out the principles of sample size estimation as well as to emphasize its importance not only in general but also in medical rehabilitation research. PMID- 17123223 TI - A prospective on stem cell research. AB - Stem cell research has stimulated considerable recent interest, but the concepts are old. Nevertheless, our understanding of the basic biology of different stem cell systems is poor. Many questions remain to be answered: How can we recognize stem cells? Are the underlying control mechanisms common to different types of stem cell, the so-called stemness concept, or is the control of self-renewal and commitment distinct in different stem cell types? What is the significance of differences in stem cells from different species? Do stem cells from somatic tissues really show plasticity with an ability to generate cells from distinct lineages, or are the observed examples consequences of experimental artifact, or rare events of no physiological significance? Do genetic mutations in the genes controlling stem cell self-renewal and differentiation lie at the heart of carcinogenesis? Answers to these and related questions now offer exciting future possibilities for both basic biology and medicine. PMID- 17123224 TI - Sources, derivation, and culture of human embryonic stem cells. AB - Human embryonic stem cells (hESCs) are immortal cells capable of perpetual self renewal in culture while maintaining their undifferentiated state, high telomerase activity, normal karyotype, and specific pattern expression of embryonic surface markers and pluripotent transcription factors such as Oct-4 and Nanog. Since their first derivation in 1998, hundreds of hESC lines have been derived and characterized. Normal surplus embryos from IVF programs are the main source for the derivation of hESC lines but cell lines from poor-quality discarded embryos or embryos carrying genetic defects following preimplantation genetic diagnosis were also isolated. Such isolation is usually accomplished by either mechanical or immunosurgical removal of the trophectoderm and culture of the inner cell mass on inactivated feeder cells. In light of the future need for clinical-grade cells, the subject of defining specific culture conditions has been addressed widely. Indeed, derivation and maintenance of hESCs without feeder cells and in media free of animal products have been attained recently. This well defined culture system may facilitate research and clinical applications, and use the remarkable potential of these exceptional cells to its fullest in both the laboratory and the clinic. PMID- 17123225 TI - Embryonic germ cells: when germ cells become stem cells. AB - Embryonic germ cells (EGCs) are pluripotent stem cells derived from primordial germ cells (PGCs). PGCs are progenitors of adult gametes, which diverge from the somatic lineage between late embryonic to early fetal development. First derived in the mouse, EGCs have also been derived from human, chicken, and pig. As pluripotent stem cells, EGCs demonstrate long-term self-renewal via clonal expansion in an undifferentiated state, and differentiate in vitro to form embryoid bodies containing cells that represent all three germ layers as well as mixed cell populations of less differentiated progenitors and precursors. This is also demonstrated in vivo by their formation into experimentally induced teratocarcinomas following transplantation. Furthermore, mice, pig, and chicken EGCs have also been shown to contribute to experimentally produced chimeric animals, including germline transmission. Importantly, EGCs demonstrate normal and stable karyotypes as well as normal patterns of genomic imprinting, including X-inactivation. Transplantation studies have begun in a variety of models in hopes of defining their potential use to treat a wide variety of human conditions, including diabetes and urological and neurological disorders. PMID- 17123226 TI - Human embryonic stem cells as a model for trophoblast differentiation. AB - The earliest stages of human development profoundly influence the success of pregnancy and maternal as well as fetal health. One of the most critical developmental processes is the specification of the trophoblast lineage and the formation of the placenta, yet this is the stage that is most difficult to study because of the general inability to do in vitro studies with human embryos or conduct in vivo clinical research during the initial weeks of pregnancy. Embryonic stem cells potentially provide a novel surrogate for the undifferentiated pluripotent cells of the preimplantation embryo, and current research indicates that human embryonic stem cells are capable of differentiation to the trophoblast lineage under the influence of diverse soluble, transcriptional, and environmental cues. This review summarizes current areas of research in these different approaches to the study of placental cell differentiation and suggests avenues of investigation that are both unique and promising in this area. PMID- 17123227 TI - Embryonic stem cells as a potential source of gametes. AB - Embryonic stem cells (ESCs) have a remarkable capacity for pluripotency; that is, they are capable of differentiating into all types of cell lineages. Recent studies have reported that successful differentiation of mouse ESCs into primordial germ cells (PGCs) as well as into mature male and female gametes can be achieved in vitro and in vivo. In addition, human ESCs can be differentiated into PGCs. The differentiation of ESCs into germ cells of various stages appears to be a spontaneous and quick process, probably due to the nature of ESCs themselves and the microenvironment of the culture conditions that favor this process. Although the functionality of these ESC-derived gametes remains to be established, derivation of both male and female gametes from ESCs raises the possibility of using these gametes to gain a better understanding of basic reproductive biology and, in particular, in conjunction with nuclear transfer technology, to extend the potential for therapeutic cloning and the treatment for infertility. We review the current knowledge on the potential of ESCs to differentiate into both male and female gametes. PMID- 17123228 TI - Genomic approaches to early embryogenesis and stem cell biology. AB - Large-scale systematic gene expression analyses of early embryos and stem cells provide useful information to identify genes expressed differentially or uniquely in these cells. We review the current status of various approaches applied to preimplantation embryos and stem cells: expressed sequence tag, serial analysis of gene expression, differential display, massively parallel signature sequencing, DNA microarray (DNA chip) analysis, and chromatin-immunoprecipitation microarrays. We also discuss the biological questions that can only be addressed by the analysis of global gene expression patterns, such as so-called stemness and developmental potency. As the emphasis now shifts from expression profiling to functional studies, we review the genome-scale functional studies of genes: expression cloning, gene trapping, RNA interference, and gene disruptions. Finally, we discuss the future clinical application of such methodologies. PMID- 17123229 TI - Fetal stem cells: betwixt and between. AB - Fetal stem cells can be isolated not only from fetal blood and hemopoietic organs in early pregnancy, but from a variety of somatic organs as well as amniotic fluid and placenta throughout gestation. Fetal blood is a rich source of hemopoietic stem cells, which proliferate more rapidly than those in cord blood or adult bone marrow. First-trimester fetal blood, liver, and bone marrow also contain a population of mesenchymal stem cells, which appear to be more primitive with greater multipotentiality than their adult counterparts. Fetal stem cells may thus represent an intermediate cell type in the current debate focusing on dichotomized adult versus embryonic stem cells, and thus prove advantageous as a source for downstream cell therapy applications. They have also been implicated in fetomaternal trafficking in pregnancy, and in long-term microchimerism in postreproductive women. PMID- 17123230 TI - In utero stem cell transplantation. AB - Successful in utero stem cell transplantations with hematopoietic or other stem cells should represent a major step forward in the management of patients with congenital, hematological, metabolic, and immunological disorders. The possibility of performing cell transplantations with stem cells across histoincompatibility barriers without chemotherapy has great potential for both pre and postnatal transplantations. The present article includes an overview of this topic with special reference to different animal models and the experience in humans in regard to fetal stem cell transplantations. This area of research holds great promise for the future, and development of efficient intrauterine stem cell treatment may enable the relative new speciality of fetal medicine to progress from a mainly diagnostic to a therapeutic medical subject. PMID- 17123231 TI - Stem cells from umbilical cord blood. AB - The study of hematopoiesis, the generation of blood cell lines throughout life, has provided conceptual, experimental, and therapeutic approaches useful to all stem cell biologists. From a clinical perspective, no other area of stem cell biology has been applied as successfully as has transplantation of bone marrow and cord blood for the treatment of blood diseases. In the last few years, research in stem cell biology has expanded rapidly to include the study of stem cells from embryonic, fetal, and various adult tissues, engendering novel perspectives regarding the identity, origin, and full therapeutic potential of tissue-specific stem cells. Rather than focusing on the use of cord blood stem cells for reconstitution of bone marrow, this article reviews the biology of stem cells found in the cord blood in the context of cell plasticity and their therapeutic potential for repair of the nervous system. PMID- 17123232 TI - Testicular stem cells. AB - Spermatogenesis originates from a constantly renewing population of testicular stem cells (TSCs). The existence of TSCs offers clinically relevant options for preservation and reestablishment of male fertility. Therefore, it is essential to understand the fundamental biology of TSCs and the basic mechanisms of self renewal, differentiation, and regulation. Although fundamental knowledge of the biology of TSCs is limited, a number of markers have been identified recently. This article reviews the current knowledge of the biology, proliferation, and regulation of TSCs. In addition, new approaches to preserve male fertility are described. TSC transplantation and grafting can be applied to generate spermatozoa from TSCs. These techniques could therefore prove important for fertility restoration in young cancer patients. Based on this exciting research, this review also focuses on current knowledge and future options for cryopreservation, culture, selection, and transfection of TSCs. PMID- 17123233 TI - Pluripotency in adult stem cells: state of the art. AB - For many years, it has long been known that stem cells derived from adult tissues maintain the capacity for self-renewal and differentiation into multiple cell types that are characteristic of the tissue of origin. Recent studies have shown new evidence that several tissues may contain cells capable of generating differentiated cells beyond their own tissue boundaries, defining a process termed stem cell plasticity. The pluripotency of adult stem cells have evoked significant excitement over the possibility of novel functional uses of stem cells, with the final purpose to develop new and more effective treatment strategies. However, despite the number of promising studies describing the plasticity of adult stem cells, many questions remain to be answered. In this article, we critically review the current state of the art in the field of adult stem cells, focussing on the present understanding of the concepts of stem cell pluripotency and plasticity. PMID- 17123234 TI - [Obstructive airway diseases due to occupation and profession]. AB - Prevalences of obstructive airway diseases, i.e. of bronchial asthma and COPD, are in the range of 5% in our populations. Approximately every 10th of these cases is caused by occupational agents. Due to different pathomechanisms and legal definitions allergic disorders have to be separated from irritant disorders. A further entity represents isocyanate induced diseases (caused by hardeners, agents for manufacturing of plastics etc.). The latter mainly behave as irritants but may also elicit allergic reactions. Among causative occupational allergens flour, food and feed products and latex dominate; among irritative agents hairdresser material, welding and cutting fumes and varnishes are most important. In most cases prognosis is poor; it depends on duration of exposure to causative agents. This emphasizes the need of early diagnosis and individual preventive measures. PMID- 17123235 TI - [Treatment of chemotherapy-induced nausea and vomiting with 5-hydroxytryptamine type 3 receptor antagonists]. AB - BACKGROUND AND OBJECTIVE: 5-hydroxytryptamine 3 (5-HT3) receptor antagonists have proved to be highly efficacious in the prevention and treatment of chemotherapy induced nausea and vomiting (CINV). However, several questions remain concerning the relative efficacy of various approved anti-emetics, especially with respect to dosage, duration and timing of administration, as well as differences in toxicity profiles. Thus it seemed appropriate to assess the current therapeutic results in routine daily practice, when applying antiemetic therapy according to established guidelines. PATIENTS AND METHODS: Within LINO e.V., a group of medical oncologists in private ambulant practice (LINO: Association of private practice medical oncologists in Bavaria), a total of 738 cycles of moderately or highly emesis-inducing chemotherapy were randomly assigned to three treatment options: (1)granisetron 1 mg (GRA1). (2) 3 mg (GRA3), and (3) ondansetron 8 mg (OND8), each combined with 8 mg dexamethasone. Incidence and severity of acute (day 1) and delayed CINV (day 2-5) and the subjective assessment of efficacy were documented on questionnaires filled in by the patients. RESULTS: All of the three regimens adequately prevented vomiting in the majority of the patients. However, all measured effects showed an uniform trend towards slightly decreased efficacy with 8 mg of ondansetron. These differences were predominantly detected with respect to delayed CINV on days 2 to 5, especially after chemotherapy with anthracyclin/cyclophosphamide combination therapy. CONCLUSION: Granisetron, at two different dosage regimens, and ondasetron showed adequate reduction in chemotherapy-induced nausea and vomiting, but ondansetron worked slightly less so. PMID- 17123237 TI - [Acute gastrointestinal bleeding caused by perforation of a Greenfield caval filter into the duodenum]. AB - HISTORY AND CLINICAL FINDINGS: A 77-year-old woman was admitted to a nearby hospital because of acute upper gastrointestinal bleeding and collapse. A Greenfield caval filter had been implanted nine years before admission because of pulmonary embolism. INVESTIGATIONS: Gastroduodenoscopy showed two hooks of the caval filter having penetrated the duodenum. The diagnosis was confirmed by an abdominal CT scan. TREATMENT AND COURSE: The patient was transferred to our hospital for surgical removal of the cava filter, which was done through the right-flank retroperitoneal approach. She had an uneventful recovery and was discharged from the hospital on the 7th postoperative day. CONCLUSIONS: Acute upper gastrointestinal bleeding caused by by a Greenfield cava filter perforating the duodenum is an extremely rare complication. But in case of acute gastrointestinal bleeding in a patient with an implanted caval filter or vascular prosthesis this should be considered and the filter removed surgically. PMID- 17123238 TI - [Fulvestrant]. PMID- 17123239 TI - [Nosocomial pneumonia: therapy]. PMID- 17123240 TI - [Control of infection in gastrointestinal endoscopy]. PMID- 17123241 TI - [Definitions and recommendations for palliative sedation]. PMID- 17123242 TI - [Presence of obligations for continuing medical education empower chief physicians in continuing education]. PMID- 17123243 TI - Guidelines for resident training in veterinary clinical pathology. II. Hematology. AB - BACKGROUND: The Education Committee of the American Society for Veterinary Clinical Pathology (ASVCP) identified a need for improved structure and guidance in training residents in clinical pathology. To begin to meet this need, guidelines for training in clinical chemistry were published in 2003. OBJECTIVE: The goal of this report is to define learning objectives and competencies in hematology, including coagulation and immunohematology. METHODS: These guidelines were developed and written with the input of ASVCP Education Committee members and peer experts. RESULTS: The primary objectives of training in hematology are: 1) to accrue a thorough, extensive, and relevant knowledge base of the types, principles, and properties of hematology tests and concepts of pathophysiology in animals; 2) to develop abilities to reason, think critically, communicate effectively, and exercise judgment in hematologic data interpretation and investigative problem-solving; and 3) to acquire technical and statistical skills important in hematology and laboratory operations. We also provide options and ideas for training activities and identify hematology resources useful for clinical pathology faculty and staff, training program coordinators, and residents. CONCLUSIONS: The guidelines define expected competencies that will help ensure proficiency, leadership, and the advancement of knowledge in veterinary hematology and provide a useful framework for didactic and clinical activities in resident-training programs. The learning objectives can readily be adapted to institutional and individual needs, interests, goals, and resources. PMID- 17123244 TI - Architectural patterns in cytology: correlation with histology. AB - Cellular arrangements are highly relevant in the evaluation of histologic sections, but cytologic specimens are not thought to accurately reflect the same patterns. Only occasionally do cytologic descriptions comprise, apart from cytoplasmic and nuclear features, architectural patterns. Architectural arrangement of cells in cytologic specimens, however, may mirror the corresponding arrangement of cells in histologic specimens provided the cytologic preparations are collected and prepared appropriately. Recognition and understanding of the different types of cell arrangements that may be seen in cytologic specimens can provide important information for obtaining a definitive diagnosis, particularly for neoplastic lesions. The goal of this review is to describe the most common architectural patterns observed in cytologic specimens and to draw correlations with similar histologic arrangements. PMID- 17123245 TI - Subgroups of canine antinuclear antibodies in relation to laboratory and clinical findings in immune-mediated disease. AB - BACKGROUND: Autoimmune system diseases in dogs are commonly referred to as systemic lupus erythematosus (SLE), with a positive antinuclear antibody (ANA) test as a hallmark. In human patients, other systemic ANA-positive diseases with overlapping diagnostic features, referred to as SLE-related diseases, are described. OBJECTIVES: The objective of this study was to investigate whether different patterns of ANA reactivity represent different systemic autoimmune diseases in dogs. METHODS: Dogs with serum positive for ANA by indirect immunofluorescence (IIF-ANA, titer > or = 1:100) (n = 56) were identified retrospectively from the patient population at the Department of Small Animal Clinical Sciences, Swedish University of Agricultural Sciences. Dogs were grouped on the basis of ANA staining patterns, and the results of immunodiffusion tests. Clinical, hematologic, serum biochemical, radiologic, and pathologic examinations were described for each group. RESULTS: Dogs with a chromosomal-positive, homogeneous ANA staining pattern (n = 14) had clinical signs involving multiple organ systems; 8 dogs were anemic. Dogs with a speckled IIF-ANA staining pattern (n = 42) primarily had clinical signs of musculoskeletal disorders, fatigue and fever. Precipitating antibodies by immunodiffusion were found only in dogs with a speckled IIF-ANA staining pattern and comprised 4 different subgroups based on antigen specificity. CONCLUSIONS: In dogs with homogeneous IIF-ANA staining, SLE is a probable diagnosis because of the diversity of clinical manifestations and autoantibody reactivity against chromosomal antigens. Dogs with a speckled IIF ANA pattern may have SLE-related diseases, which, in turn, may be correlated with different immunodiffusion subgroups. These syndromes had overlapping clinicopathologic features, as described for human patients. PMID- 17123246 TI - Serum chemistry values for free-ranging ringed seals (Pusa hispida) in Svalbard. AB - BACKGROUND: Diseases and abnormal physiologic conditions can alter the concentrations of enzymes, metabolites, minerals, and hormones in the blood of animals. The ringed seal (Pusa hispida) has been selected as a key species for environmental monitoring, but information on disease and health parameters for this species is scarce. OBJECTIVES: The aim of the study reported here was to obtain serum chemistry reference intervals for free-ranging ringed seals in Svalbard, and then to evaluate serum chemistry values in relation to age, body condition, and sex. METHODS: Blood samples were collected after death from ringed seals in Wijdefjorden and Billefjorden, Svalbard (2002-2003; n = 75). Serum was analyzed for 24 selected serum chemistry parameters (enzymes, protein, metabolites, minerals, and cortisol). RESULTS: Compared with younger or older animals, seals between 7 and 16 years of age had larger variations in the activities of alanine transaminase, aspartate transaminase, lactate dehydrogenase, and creatine kinase (CK). Animals classified as having low body condition status had more variation in the serum activity of these enzymes, compared with that in animals with higher condition scores. Serum cortisol concentration was higher in young animals (1-5 years) than in older animals. Serum CK activity was higher in males than in females. CONCLUSION: The data reported here may be useful in monitoring the health of ringed seals and for tracking the impact of environmental changes in the Arctic. PMID- 17123247 TI - A retrospective study of aplastic pancytopenia in the dog: 9 cases (1996-2003). AB - BACKGROUND: Aplastic pancytopenia is defined by the presence of pancytopenia in blood and a hypocellular bone marrow with the hematopoietic space replaced by adipose tissue. Several causes of acquired aplastic pancytopenia are known; however, in some cases, an underlying cause is never determined. OBJECTIVE: The objective of this retrospective study was to identify the incidence, potential causes, and outcome of aplastic pancytopenia in dogs. METHODS: Bone marrow cytologic and core biopsy reports were reviewed to identify dogs diagnosed with aplastic pancytopenia between July 1, 1996 and June 30, 2003. Four-hundred eighty six bone marrow reports that included aspirate and core biopsy evaluations were reviewed. Signalment, treatment given, previous and current disease conditions, clinical signs of disease, clinical laboratory data, therapy, response to therapy, and survival time were recorded. RESULTS: Nine dogs (1.85% of bone marrow samples reviewed) met the criteria for inclusion. Two dogs (22%) had associated diseases that included monocytic ehrlichiosis and Sertoli cell tumor. In 7 dogs (78%), the cause of aplastic pancytopenia could not be definitively determined, although an idiosyncratic drug reaction to griseofulvin was suspected in 1 of the dogs. The median age of dogs diagnosed with aplastic pancytopenia was 3.2 years, and apparent breed or sex predilection was not identified. Median HCT, total WBC count, and platelet count on the day of presentation were 21.8%, 1.0 x 10(3)/microL, and 2.0 x 10(3)/microL, respectively. Six of 9 dogs diagnosed with aplastic pancytopenia died or were euthanized within 21 days. Two dogs had complete hematologic recovery. One dog was living 3 years after diagnosis, but hematologic recovery was never documented. CONCLUSIONS: Aplastic pancytopenia is diagnosed infrequently and idiopathic aplastic pancytopenia may account for up to 67% or more of canine cases. Although the prognosis is guarded, some dogs with aplastic pancytopenia recover. PMID- 17123248 TI - Cloning and expression of the gene encoding the major surface protein 5 (MSP5) of Anaplasma phagocytophilum and potential application for serodiagnosis. AB - BACKGROUND: Anaplasma phagocytophilum (formerly known as the human granulocytic ehrlichia, Ehrlichia equi and Ehrlichia phagocytophila) is an obligate intracellular organism causing clinical disease in humans and various species of domestic animals. OBJECTIVES: The objectives of this investigation were to sequence and clone the major surface protein 5 (MSP5) of A phagocytophilum and to evaluate the suitability of this antigen in the serologic diagnosis of anaplasmosis in humans and dogs. METHODS: The msp5 gene of A phagocytophilum was sequenced, cloned, and expressed in Escherichia coli. The predicted amino acid sequence homology of the various MSP5/major antigenic protein 2 orthologs was compared among various Anaplasma and Ehrlichia species. Recombinant MSP5 of A phagocytophilum was used in an ELISA to detect antibodies in serum samples from humans and dogs infected with the organism. RESULTS: Serum samples from 104 individuals previously diagnosed with A phagocytophilum infection, as well as samples from clinically healthy humans, were tested. In addition, multiple samples from 4 dogs experimentally infected with 2 different geographic isolates of A phagocytophilum and 5 dogs naturally infected with a Swiss isolate were tested using ELISA. Using this group of immunofluorescent antibody test-positive and immunofluorescent antibody test-negative samples, we found the overall agreement between assays to be >90%. CONCLUSIONS: These results indicate that recombinant MSP5 has potential for use as a diagnostic test antigen to detect infection with A phagocytophilum in both dogs and humans. However, sequence similarities among orthologs of MSP5 in related species of anaplasma and ehrlichia suggest that cross-reactivity among these pathogens is likely if the entire peptide is used as a test antigen. PMID- 17123249 TI - Assessment of the use of plasma and serum chloride concentrations as indirect predictors of serum bromide concentrations in dogs with idiopathic epilepsy. AB - BACKGROUND: Bromide (BR) administration causes pseudohyperchloremia when plasma or serum chloride (Cl-) concentrations are determined with commonly available automated analytical assays. In humans receiving BR, it has been previously demonstrated that the plasma Cl- concentration is a useful indirect estimator of the measured BR concentration. OBJECTIVE: The objective of this study was to determine if the magnitude of pseudohyperchloremia seen in epileptic dogs treated with BR could be used as a predictor of the measured serum BR concentration. METHODS: Plasma and serum Cl- concentrations, analyzed by ion-specific electrode (ISE) and colorimetric techniques, and serum BR concentrations, determined using the gold-trichloride assay, were simultaneously determined in 88 blood samples from dogs with idiopathic epilepsy that were treated with BR. RESULTS: For all methods used to quantify Cl- concentrations, there were significant (P < .0001) linear relationships between BR and Cl- concentrations. Linear relationships between BR and Cl- concentrations were significantly different (P < .0001) between blood samples from dogs obtained during routine therapeutic monitoring and those obtained during emergency hospital admissions. Calculated 95% prediction intervals for future values of BR using measured Cl- concentrations contained considerable error. Plasma Cl- values determined with ISE generally provided the best prediction of serum BR concentrations. Agreement between the measured BR and Cl- using all Cl- assay techniques was moderate, but was statistically significant only when Cl- was assayed in plasma using one ISE method. CONCLUSIONS: The pseudohyperchloremia observed in epileptic dogs receiving BR is an inadequate indirect estimator for the measured BR concentration, although in certain clinical situations identified through construction of a clinical decision tree, the measured Cl- value can be used to guide general therapeutic decisions regarding alterations in BR therapy. Optimal tailoring of BR therapy in dogs with idiopathic epilepsy should be based on results of therapeutic monitoring of BR concentrations. PMID- 17123250 TI - Hematology and plasma chemistry reference intervals for cultured shortnose sturgeon (Acipenser brevirostrum). AB - BACKGROUND: The shortnose sturgeon, Acipenser brevirostrum, is an imperiled species distributed along the Atlantic coast of North America. Interest in replenishing wild stocks with hatchery-reared fish has created a need for accurate hematologic and biochemical reference intervals to evaluate the health of both fish raised in aquaculture systems and fish in the wild. OBJECTIVES: The objective of this study was to generate hematologic and biochemistry reference intervals for healthy shortnose sturgeon. METHODS: Blood samples were collected in heparinized tubes from 77 shortnose sturgeon raised in flow-through aquaculture systems. Whole blood and plasma samples were analyzed for hematologic and biochemical variables using standard techniques. Reference intervals were calculated as the central 95% (percentile) of data. RESULTS: Hematologic reference intervals (n = 46) were as follows: PCV 26-46%, hemoglobin 5.7-8.7 g/dL, MCV 307-520 fL, MCH 65.9-107.1 pg, MCHC 15-30 g/dL, plasma proteins (refractometry) 2.8-6.0 g/dL, RBC count 0.65-1.09 x 10(6)/microL, total WBC count 28,376-90,789/microL, small lymphocytes 9063-56,656/microL, large lymphocytes 2122-10,435/microL, neutrophils 3758-33,592/microL, monocytes 0-7137/microL, eosinophils 0-1544/microL, thrombocyte-like cells 6863-23,046/microL, thrombocytes 32,205-122,179/microL, and neutrophil:lymphocyte ratio 0.068-1.026. Plasma chemistry reference intervals (n = 77) were as follows: total protein 2.7 5.3 g/dL, albumin 0.8-1.7 g/dL, globulins 1.8-3.7 mg/dL, creatinine 0-1.4 mg/dL, total bilirubin 0-0.1 mg/dL, alkaline phosphatase 47-497 U/L, aspartate aminotransferase 90-311 U/L, sodium 124-141 mmol/L, potassium 2.9-3.7 mmol/L, chloride 106-121 mmol/L, calcium 6.6-12.1 mg/dL, magnesium 1.6-2.3 mg/dL, phosphorus 5.1-8.1 mg/dL, glucose 37-74 mg/dL, cholesterol 42-133 mg/dL, and osmolality 232-289 mOsm/kg. CONCLUSION: Reference values reported here will be useful for the early detection, identification, and monitoring of disease and sublethal conditions in cultured shortnose sturgeon. PMID- 17123251 TI - Serum ionized calcium in dogs with chronic renal failure and metabolic acidosis. AB - BACKGROUND: Chronic renal failure (CRF) is a common disease in dogs, and many metabolic disorders can be observed, including metabolic acidosis and calcium and phosphorus disturbances. Acidosis may change the ionized calcium (i-Ca) fraction, usually increasing its concentration. OBJECTIVE: In this study we evaluated the influence of acidosis on the serum concentration of i-Ca in dogs with CRF and metabolic acidosis. METHODS: Dogs were studied in 2 groups: group I (control group = 40 clinically normal dogs) and group II (25 dogs with CRF and metabolic acidosis). Serum i-Ca was measured by an ion-selective electrode method; other biochemical analytes were measured using routine methods. RESULTS: The i-Ca concentration was significantly lower in dogs in group II than in group I; 56% of the dogs in group II were hypocalcemic. Hypocalcemia was observed in only 8% of dogs in group II when based on total calcium (t-Ca) concentration. No correlation between pH and i-Ca concentration was observed. A slight but significant correlation was detected between i-Ca and serum phosphorus concentration (r = .284; P = .022), as well as between serum t-Ca and i-Ca concentration (r = .497; P < .0001). CONCLUSION: The i-Ca concentration in dogs with CRF and metabolic acidosis varied widely from that of t-Ca, showing the importance of determining the biologically active form of calcium. Metabolic acidosis did not influence the increase in i-Ca concentration, so other factors besides acidosis in CRF might alter the i-Ca fraction, such as hyperphosphatemia and other compounds that may form complexes with calcium. PMID- 17123252 TI - Fractal dimension of canine mammary gland epithelial tumors on cytologic smears. AB - BACKGROUND: Fractal geometry is a tool that can be used for describing, modeling, analyzing, and processing irregular and complex figures. Past investigations in medicine have revealed that fractal analysis could also be applied in tumor pathology to characterize irregular boundaries of the nuclei of tumor cells. OBJECTIVE: The aim of this study was to define whether the fractal dimension parameter could be used on cytologic specimens to differentiate benign from malignant canine mammary gland epithelial tumors. METHODS: The fractal dimension of nuclear surface was determined by computer-assisted morphometry on Hemacolor stained cytologic smears obtained by fine needle aspiration of normal canine mammary gland epithelial cells, and cells in mammary adenomas, tubulopapillary carcinomas, solid carcinomas, and anaplastic carcinomas. Data were analyzed by Mann-Whitney U test. RESULTS: Significant differences (P <.001) were observed in mean fractal dimension among all tumor types and in comparison with normal canine mammary gland epithelial cells (except for the fractal dimension between solid carcinomas and anaplastic carcinomas). CONCLUSION: The morphometric parameter, fractal dimension, could help in the diagnostic discrimination between benign and malignant canine mammary gland epithelial tumors on cytologic specimens. PMID- 17123253 TI - Basal cell tumor or cutaneous basilar epithelial neoplasm? Rethinking the cytologic diagnosis of basal cell tumors. AB - A 1-cm-diameter, red, raised, cutaneous mass over the dorsal surface of the left third metacarpal of a 6-year-old neutered male yellow Labrador Retriever was aspirated. The aspirate contained cohesive clusters of cells, some containing cells with increased pleomorphism. Cellular debris (some keratinized), melanin, large numbers of macrophages, a few neutrophils, and fibroblasts were also observed. The cytologic interpretation was malignant neoplasia with histiocytic inflammation. Differentials included a carcinoma or, given the melanin pigment and variable morphology of the cells, possibly malignant melanoma. Histologically, the tumor was diagnosed as a basal cell epithelioma. Neoplasms that once were lumped into the broad histologic diagnosis of basal cell tumors have since been split into distinct entities, dependent on evidence of differentiation into epidermis, trichofollicular epithelium, or sweat or sebaceous glands. Although histologic reclassification has resulted in removal of most of these entities from the original basal cell tumor category, a cytologic diagnosis of basal cell tumor continues to be used to represent the large, heterogeneous group of epidermal, trichofollicular, and adnexal skin tumors with basal cell characteristics. The case in this report demonstrates the heterogeneity of neoplasms that may be diagnosed cytologically as basal cell tumors and supports the need for cytologic criteria and nomenclature that better reflect potential variation in tissue differentiation. PMID- 17123254 TI - Chronic eosinophilic leukemia in a cat: cytochemical and immunophenotypical features. AB - A 3-year-old, male, domestic shorthaired cat was presented with a 3-day history of anorexia and depression. The cat was moderately dehydrated, had pale, slightly icteric, mucous membranes, oral ulcerations, and mild hepatosplenomegaly. A feline leukemia virus (FeLV) antigen test was positive. CBC results obtained at initial presentation included severe normocytic, normochromic, nonregenerative anemia, severe thrombocytopenia, and marked leukocytosis (>100,000/microL) with 77% eosinophils. After 15 days of treatment with prednisone and doxycycline, the cat had persistent severe nonregenerative anemia (HCT 3.4%), thrombocytopenia (28,000/microL), and extreme eosinophilia (total eosinophils, 123.1 x 10(3)/microL; segmented 103.0 x 10(3)/microL; immature 20.1 X 10(3)/microL). Cytologic examination of aspirates from bone marrow, liver, lymph nodes, and spleen revealed a predominance of mature and immature eosinophils, many with dysplastic changes. The M:E ratio was 96.4. On histopathologic examination, multiple organs were infiltrated by eosinophilic granulocytes. Neoplastic cells in blood and bone marrow stained positive for alkaline phosphatase and were negative for myeloperoxidase, chloroacetate esterase, and alpha-naphthyl acetate esterase. On flow cytometric analysis of peripheral blood, the neoplastic cells were positive for CD11b and CD14. These findings were consistent with chronic eosinophilic leukemia. To our knowledge, this is the first report of chronic eosinophilic leukemia in a cat associated with naturally acquired FeLV infection, in which flow cytometry was used to characterize the neoplastic cells. PMID- 17123255 TI - Congenital Pelger-Huet anomaly in a horse. AB - A 1.5-year-old male Arabian horse was referred to the Louisiana State University Veterinary Teaching Hospital and Clinic for an open deep laceration involving two thirds of the right trunk. The initial CBC results included an inflammatory leukogram, characterized by a marked degenerative left shift consisting of only immature band neutrophils (7500/microL, reference interval 0-100/microL) with toxic changes and no segmented neutrophils (0/microL, reference interval 2700 6700/microL). On abdominal ultrasonography, free abdominal fluid was found and collected for analysis. Abdominal fluid had a marked increase in total nucleated cells (40,600 cells/microL) consisting of 74% nondegenerate neutrophils that all were hyposegmented, with mature condensed chromatin. Re-evaluation of neutrophil morphology on the initial blood smear confirmed hyposegmentation and mature condensed chromatin, similar to that observed in cells in the abdominal fluid. A diagnosis of Pelger-Huet anomaly (PHA) was made in this colt. Congenital PHA was documented on the basis of persistent neutrophil hyposegmentation on serial blood smears, ruling out of acquired causes of PHA, and findings of similar neutrophil hyposegmentation on blood smears from the colt's sire and the sire's siblings. To our knowledge, this is the first reported case of congenital PHA in a horse. PMID- 17123256 TI - Blood smear from a cat: features to "dys"cover. AB - A 4-year-old, spayed female, domestic shorthair cat was presented for lethargy, nonregenerative anemia, and inappetence. Results of a CBC included macrocytic, normochromic, nonregenerative anemia and a glucocorticoid-associated leukogram. On blood smear examination, neutrophils had abnormal features including hyposegmentation and a diffuse chromatin pattern with nuclear filament formation and nuclear blebbing. Microscopic examination of a roll preparation of bone marrow revealed hypolobulated megakaryocytes with asynchronous maturation of nuclei. The granulocytic to erythrocyte (G:E) ratio was 76. Segmented neutrophils had asynchronous maturation and dysplastic features. The entire erythroid lineage was markedly decreased for the degree of anemia and rare dysplastic features were noted in erythroid precursor cells. The interpretation of bone marrow findings was erythroid hypoplasia, megakaryocytic dysplasia, and granulocytic hyperplasia with dysplasia. Histopathologic examination of a bone marrow core sample also revealed myeloid hyperplasia and erythroid hypoplasia. The result of a direct immunofluorescence assay for FeLV performed on the bone marrow roll preparation was positive. A diagnosis of dysmyelopoiesis associated with FeLV infection was made. This case was unique in that the dysplastic changes occurred in cell lines that did not have associated cytopenias. The dysmyelopoiesis most closely resembled myelodysplastic syndrome with refractory cytopenia (MDS-RC); however, secondary dysmyelopoiesis could not be ruled out. PMID- 17123257 TI - Gallbladder aspirate from a dog. AB - A 7-year-old, male, castrated, Labrador Retriever with a history of pancreatitis and inflammatory bowel disease presented for vomiting and anorexia. Serum biochemistry findings were indicative of cholestasis, hepatocellular insult, and decreased hepatic function. Ultrasound examination showed sediment and gas within the gallbladder, and a diagnosis of emphysematous cholecystitis was made. Emergency gallbladder resection was performed. Cytologic examination of bile fluid collected at surgery showed a mixed population of bacteria (bactibilia) together with fungal organisms consistent with Cyniclomyces guttulatus (previously known as Saccharomycopsis guttulatus). Similar fungal organisms were seen on a fecal smear. Bacteria cultured were normal gastrointestinal flora, supporting ascending infection; the fungal organisms were interpreted as incidental. Histopathology of the gallbladder indicated active (suppurative) and chronic (lymphocytic) cholecystitis and sections of liver tissue had evidence of chronic liver disease. A positive liver culture indicated concurrent bacterial hepatitis or cholangiohepatitis. Despite supportive care, the dog continued to decline and was euthanized 30 days later. Necropsy results confirmed end stage liver disease, but an initiating cause was not found. This case highlights the role of bactibilia in the development of acute cholecystitis and the unique cytologic appearance of C guttulatus as an incidental finding in bile fluid. PMID- 17123258 TI - Transtracheal wash from a puppy with respiratory disease. AB - A 4-month-old, intact male Boxer puppy was presented to the Animal Emergency and Critical Care Services of South Florida because of nasal discharge, dehydration, dyspnea, and coughing. The dog had been diagnosed with intestinal parasites and kennel cough approximately 10 days before presentation. Lateral and ventrodorsal radiographs of the thorax revealed an increased bronchointerstitial pattern throughout the lungs. A transtracheal wash was performed. On cytologic examination of direct, Wright-Giemsa-stained smears, small basophilic coccoid structures (0.3-0.9 microm in diameter) were observed in low to moderate numbers within neutrophils and adherent to epithelial cells. The small size of the organisms raised suspicion for Mycoplasma. Culture of the transtracheal wash fluid resulted in growth of a Mycoplasma sp. The patient was treated with enrofloxacin and amoxicillin/clavulanate and made a full recovery. Recognizing Mycoplasma in transtracheal washes could aid in recommending the appropriate culture media or immunologic techniques, which could result in an accurate diagnosis of mycoplasmosis. PMID- 17123259 TI - [The pathogenetical mechanisms of glomerular proteinuria.]. AB - Although the pathologic mechanisms responsible for glomerular proteinuria have not been completely clarified, yet, it has become evident that structural and functional abnormalities of podocytes play an early and important role. These data come from recent studies analyzing podocyte behaviour in cell culture, in depth clinical researches, the application of survey techniques on isolated glomeruli, proteomic studies on particular aspects of circulating factors. For most glomerulopathies, whether primary or secondary, podocytes are often the first target for many pathogenetic mechanisms, which, unexpectedly, have the common characteristic of inducing tropism in podocytes. Hopefully, advances in glomerular podocyte research will better define clinical diseases, which we now classify according to a static description of the histological damage, thus perhaps mistaking causes and effects. PMID- 17123260 TI - [Kidney transplant failure and retransplantation.]. AB - Transplant failure is a more and more frequent cause of end stage renal failure and dialysis. Patient survival rate after graft failure is very varied according to different reports. Better survival is mainly a consequence of good continuum of care thanks to improved interaction between dialysis and transplant center. Diabetic and elderly patients, as well as patients affected by cardiovascular disease are the subjects at higher risk: if judged clinically adequate to enter the waiting list, they should be retransplanted as soon as possible. Dialysis survival of patients with failed kidney transplant is strictly linked to adequate dialysis dose. Second transplant survival rate is higher in the case of a living donor and if the first transplant survived longer. Good immunologic match is also a condition linked to higher graft and patient survival rate. High body mass index, smoking and severe cardiovascular comorbidity should be avoided. Whether to keep low immunosuppression levels after first graft failure and whether to excise the failed kidney, even though it shows no clinical problems, are issues still under debate. Low-dose immunosuppression is not recommended since it may result in higher rate of infectious and neoplastic diseases. The failed kidney should be removed not only in the case of clinical disease, but also when the retained failed kidney is associated with chronic inflammation, as shown by high C-reactive protein levels and erythropoietin resistance. PMID- 17123261 TI - [Hyperhomocysteinemia in chronic renal failure.]. AB - Chronic renal failure (CRF) is frequently associated with increased plasma levels of homocysteine (Hcy), an amino acid that can be considered a new uremic toxin according to recent evidence. Studies on Hcy described first homocystinuria, an inherited disease characterized by high plasma Hcy levels and premature cardiovascular disease, resulting in high mortal-ity rates. Hyperhomocysteinemia was then shown to be associated with cardiovascular events both in the general population and in CRF patients. Hcy is a sulfur amino acid derived from dietary methionine, an essential amino acid. Methionine is condensed with ATP to form S adenosylmethionine (AdoMet), the universal methyl donor in transmethylation reactions. The AdoMet demethylated product is S-adenosylhomocysteine (AdoHcy), which is the direct precursor of Hcy in vivo. Hcy is toxic for the endothelium, it enhances vascular smooth muscle cell proliferation, increases platelet aggregation, and acts on the coagulation cascade and fibrinolysis. Several mechanisms have been discussed to explain Hcy toxicity. Hcy levels increase as renal function declines and progresses to ESRD; the causes of hyperhomocysteinemia are still unclear. Studies in humans show that renal metabolic extraction depends on renal plasma flow; in addition, an alteration of the extrarenal metabolic clearance, depending on uremic toxins, may occur. Among the consequences of hyperhomocysteinemia in renal failure are: impaired protein methylation, with altered protein repair processes; DNA hypomethylation, with an alteration in the allelic expression of genes regulated through methylation; and protein homocysteinylation. Further, this review is dealing with the 'reverse epidemiology' issue, outlining also the main Hcy-lowering strategies. PMID- 17123262 TI - [P value and confidence intervals: reporting and interpreting the result of a clinical study]. AB - The main purpose of statistics in the analysis of clinical and epidemiological studies is to summarize data and information, as well as assess variability, trying to distinguish between chance findings and results that may be replicated upon repetition. Statistical analyses only convey the effect of chance element in data (random error). Statistics cannot control non-sampling errors concerning study design, conduct and methods adopted. At the end of the study, a result is defined statistically significant if the observed difference in the outcome variable is too large to be attributed to chance. A small P value provides evidence against the null hypothesis (of no effect), since data have been observed that would be unlikely if the null hypothesis was true. However, confidence intervals estimate separate the two data dimensions (strength of the relation between exposure and disease, and precision with which the relation is measured), and add to the hypothesis testing useful information for finding interpretation and further research. PMID- 17123263 TI - [Ultrasound investigation in Apulo-Lucano renal echography study group of the Italian Society of Nephrology: a cognitive report]. AB - The number of ultrasonography expert consultant Nephrologists is more and more increasing thanks to the contribution that this methodology has brought in both clinical and treatment fields. Up to now, a database of the ultrasonography benefits, as well as the main criteria for the interpretation of the urinary tract echographic examination has not been compiled, yet. We have therefore drawn up and distributed a questionnaire to the consultant nephrologists of the urinary tract echography study group (Apulo-Lucano division). This questionnaire is made up of 27 ultrasonography application and interpretation issues; it aims mainly at creating a common 'language' to reduce the variety of 'descriptors' currently employed by all different specialists and centres involved. 60 consultant nephrologists participated in the study, from the 29 Nephrology and Dialysis O.U.s of Puglia and Basilicata regions, where there is an active echographic service. Data collected show the key role of ultrasonography investigation for all nephrology patients, as high quality and cost efficient test procedure. Moreover, despite the fact that there are differences in echographic examination performance and interpretation, literature data show clearly that it is fundamental to follow general shared principles. The responsibility and task of those specializing in this discipline should be to allow reproduction and comparison of ultrasonographies, also among different operators and centres, and meta studies, i.e. 'a series of comparative studies', which are still very few in number. PMID- 17123264 TI - [A girl in the mirror]. AB - We report a case of situs ambiguous associated with particular vascular anomalies resulting in secondary arterial hypertension. Renal ultrasonography performed in this case has oriented diagnosis. PMID- 17123265 TI - [Commentary on 'Ethical issues in living renal transplantation']. AB - Legalizing kidney market: ethics considerations. The paper by E A Friedman and A L Friedman, advocating suitable kidney sale legislation, recently published in Kidney International, has aroused some controversy among the Italian Society of Nephrology Mailing List members (ML-SIN). A previous article reviewed the main issues and summarized Italian nephrologists' opinions. Generally speaking, ML-SIN participants were critical towards this proposal; the most widespread opinion was that trade of organs for transplant purposes is unethical and that Friedman's legislative suggestion is unlikely to succeed in Italy. To complete discussion, we report also the opinion of an authoritative Ethics expert. PMID- 17123266 TI - [Thin basement membrane and Alport syndrome]. AB - Alport Syndrome (AS) is an inherited disorder of the glomerular basement membrane (GBM) transmitted as an X-linked dominant form in approximately 80% of patients. This X-linked form is caused by mutations in the collagen type IV alfa 5 gene (COL4A5) located on chromosome X; in the remainder of the cases, the autosomal, mostly recessive form, results from mutations in the collagen type IV alfa 3 gene (COL4A3) or alfa 4 gene (COL4A4) located on the 2q. Diagnostic lesions can be observed by electron microscopy (EM) and are characterized by thinning, thickening and/or splitting of the GBM. Isolated thinning of the GBM is usually associated with isolated microhematuria, a non progressive condition which has often a familial trait. Skin biopsy has also recently emerged to be a valuable alternative for the diagnosis of X-linked AS, because the alfa 5 (COL4A5) chain is also expressed in the epidermal basement membrane. A reliable diagnosis can often be achieved through combined renal and cutaneous immunohistochemical analysis, even in patients with limited clinical signs or atypical histological findings, and/or without suggestive family history. The present case report is an example of such diagnostic dilemma, where these techniques allowed to make a diagnosis despite contradictive clinical and histological features in contrast with a positive family history of renal disease. PMID- 17123267 TI - The prognostic significance of perineural invasion in prostatic cancer biopsies: a systematic review. AB - Men with clinically localized prostate cancer are faced with a wide range of treatment options, and only Gleason grading is universally used as a histopathological prognostic factor for this disease. The significance of perineural invasion in diagnostic biopsies is controversial. Opinion about whether or not it should influence treatment decisions is currently almost equally divided. To address this, the authors performed a systematic review of studies that examine the association between perineural invasion and prostate cancer recurrence. MEDLINE, Embase, and the Web of Knowledge were searched from January 1990 to December 2005. Outcomes analyzed were the development of biochemical or clinical recurrence. Twenty-one articles on the association of perineural invasion in biopsies and prostate cancer recurrence after radical prostatectomy (n = 10) or radiotherapy (n = 11) were found but none on its significance in the context of watchful waiting. Structured data extraction was performed to allow comparisons between articles and to identify sources of heterogeneity to explain discrepancies in results. The considerable variation in study design, execution, and reporting precluded meta-analysis and quantitative risk estimation, but the weight of evidence suggested that perineural invasion in biopsies was a significant prognostic indicator, particularly in specific patient groups defined by presenting serum prostate-specific antigen levels and biopsy Gleason scores. Immediate treatment rather than watchful waiting may be more appropriate for patients with localized prostatic cancer and perineural invasion. However, the data are limited, and well-designed studies that use predefined stringent protocols are required to provide robust estimates of risk to aid in treatment planning. PMID- 17123268 TI - The burden of fatigue and quality of life in myeloproliferative disorders (MPDs): an international Internet-based survey of 1179 MPD patients. AB - BACKGROUND: Few objective data exist on the burden of fatigue and other constitutional symptoms in patients with myeloproliferative disorders (MPD). METHODS: The authors used validated instruments of fatigue and physical activity assessment during an Internet-based symptom survey of 1179 MPD patients (median age, 56 years; 41.4% men). RESULTS: The frequency of self-reporting was 80.7% for fatigue, which was substantially higher than that of pruritus (52.2%), night sweats (49.2%), bone pain (43.9%), fever (13.7%), and weight loss (13.1%). In the majority of patients, these symptoms restricted participation in both social functions and physical activity. In addition, 34.5% of patients needed assistance with activities of daily living, and 11.2% reported MPD-associated medical disability. As expected, the presence of myelofibrosis, anemia, splenomegaly, or other features associated with advanced disease favored a higher degree of fatigue. However, fatigue remained the major complaint also in polycythemia vera (84.9%) and essential thrombocythemia (72.4%); these figures were significantly higher than those of published controls (P < .0001). CONCLUSIONS: The current study identifies fatigue as the major contributor to poor quality of life in MPD, provides baseline information on constitutional symptoms, and underscores the need for the incorporation of quality of life assessment in clinical trials. PMID- 17123270 TI - Survival and toxicity differences between 5-day and weekly cisplatin in patients with locally advanced cervical cancer. AB - BACKGROUND: Cisplatin (CDDP) administration concomitant with radiotherapy (RT) for the treatment of locally advanced cervical cancer has evolved from an inpatient 5-day every 21-day regimen to a weekly outpatient regimen. This study was designed to test for differences in progression-free survival (PFS) and toxicity between the 2 regimens. METHODS: In all, 77 consecutive patients at a single institution with stage IB2-IV cervical cancer were included in this analysis (using the International Federation of Gynecologists and Obstetricians staging system). All patients were treated with CDDP, external beam RT, and 2 9 Gy high-dose-rate brachytherapy treatments. Two cohorts were compared: 1) 5-day, patients treated from 1995 to 2001 with CDDP 20 mg/m(2) x 5 days every 21 days concomitant with RT; 2) weekly, treated after May 2001 with CDDP 40 mg/m(2) weekly concomitant with RT. RESULTS: In all, 50 patients were treated with the 5 day regimen and 27 patients with the weekly regimen. There were no significant demographic differences between the groups. Overall 3-year PFS, controlling for stage, was 90% and 76% for 5-day and weekly groups, respectively (P = .01). Adjusting for stage, age, and completion of treatment, the risk of treatment failure among the weekly group was 3.46 times higher than the 5-day group (P = .02). The weekly group had a 3.43 times higher risk of developing acute toxicities than the 5-day group (P = .02) in advanced-stage patients. CONCLUSIONS: Patients who received weekly CDDP have a shorter 3-year PFS. Patients with advanced-stage cervical cancer who received weekly CDDP had significantly more acute toxicities. These data should be confirmed in a multi institutional, randomized, controlled study. PMID- 17123271 TI - Second primary cancers after prostatic irradiation: ensuring an appropriate analysis. PMID- 17123272 TI - Model for end-stage liver disease (MELD) exception for cystic fibrosis. PMID- 17123273 TI - Psychosocial needs assessment among an underserved, ethnically diverse cancer patient population. AB - Little attention has been directed towards identifying and addressing the psychosocial needs of ethnic minority and underserved cancer patients. This study describes the development of a psychosocial needs survey and patterns and predictors of need among an ethnically diverse underserved cancer patient population in Bronx, New York. A 34-item psychosocial needs assessment survey was developed to assess 4 categories of need: Informational, Practical, Supportive, and Spiritual. A total of 248 oncology outpatients (48% non-Hispanic whites, 25% African Americans; 19% Hispanic) completed the survey in oncology clinic waiting rooms. The survey demonstrated high internal consistency and face validity. Ethnicity was the sole predictor of needs (P < .02), even after controlling for education, time since diagnosis, treatment status, marital status, and age. The mean percentage of needs endorsed by African Americans, Hispanics, and non Hispanic whites respectively was 81%, 85%, 70% for Informational; 63%, 68%, 36% for Practical; 69%, 73%, 48% for Supportive; and 49%, 60%, 31% for Spiritual needs. This needs assessment offers clear directions in which to focus QOL intervention efforts among underserved and ethnic minority cancer patients. Cancer 2007. (c) 2006 American Cancer Society. PMID- 17123274 TI - Research institute for nurse scientists responds to the challenge to expand and strengthen research focused on breast cancer in African American women. AB - In an era where scientifically derived 'evidence' is used as a basis for nursing practice, it is imperative that nurses have a breadth of knowledge relative to the fundamentals of nursing science; knowledge of the current standards of nursing and medical practice; and knowledge of the characteristics, needs, concerns, and challenges of diverse consumer and patient population groups. Yet, while a significant body of 'evidence' that describes the experiences and needs of African American women across the breast care continuum has been generated, research suggests that there is a need to expand and strengthen this body of science. This report presents an overview of a decade of research focused on breast cancer among African American women and describes an initiative funded by the Susan G. Komen Breast Cancer Foundation to expand and strengthen nursing science that aims to reduce and/or eliminate excess breast cancer morbidity and mortality among African American women. Cancer 2007. (c) 2006 American Cancer Society. PMID- 17123275 TI - Improving follow-up to abnormal breast cancer screening in an urban population. A patient navigation intervention. AB - Delays in follow-up after cancer screening contribute to racial/ethnic disparities in cancer outcomes. We evaluated a patient navigator intervention among inner-city women with breast abnormalities. A full-time patient navigator supported patients using the care management model. Female patients 18 years and above, referred to an urban, hospital-based, diagnostic breast health practice from January to June 2000 (preintervention) and November 2001 to February 2003 (intervention), were studied. Timely follow-up was defined as arrival to diagnostic evaluation within 120 days from the date the original appointment was scheduled. Data were collected via computerized registration, medical records, and patient interview. Bivariate and multivariate logistic regression analyses were conducted, comparing preintervention and intervention groups, with propensity score analysis and time trend analysis to address the limitations of the pre-post design. 314 patients were scheduled preintervention; 1018, during the intervention. Overall, mean age was 44 years; 40% black, 36% non-Hispanic white, 14% Hispanic, 4% Asian, 5% other; 15% required an interpreter; 68% had no or only public insurance. Forty-four percent of referrals originated from a community health center, 34% from a hospital-based practice. During the intervention, 78% had timely follow-up versus 64% preintervention (P < .0001). In adjusted analyses, women in the intervention group had 39% greater odds of having timely follow-up (95% CI, 1.01-1.9). Timely follow-up in the adjusted model was associated with older age (P = .0003), having private insurance (P = .006), having an abnormal mammogram (P = .0001), and being referred from a hospital based practice, as compared to a community health center (P = .003). Our data suggest a benefit of patient navigators in reducing delay in breast cancer care for poor and minority populations. Cancer 2007. (c) 2006 American Cancer Society. PMID- 17123276 TI - Racial disparities in colon cancer. Primary care endoscopy as a tool to increase screening rates among minority patients. AB - Colon cancer is a condition whose far-reaching effects have been well documented nationally and within the state of South Carolina. Fortunately, the disease is amenable to both primary and secondary prevention through screening colonoscopy. Despite the conceptual simplicity of recommending colonoscopy, barriers exist to universal (or even widespread) screening. Currently the infrastructure necessary to achieve screening goals set by the American Cancer Society (ACS), the American College of Gastroenterology (ACG), and the South Carolina Department of Health and Environmental Control (DHEC) has not been established. At current rates of training gastroenterologists, the medical community will not be able to come close to achieving widespread screening. Given the discrepancy between the public health benefit of achieving the goals and the deaths that have occurred because of the resource shortfall, we propose alternative measures to screen the at-risk population for consideration. This need is most acute in the black community, in which where screening rates tend to be lower and polyps have been found to progress more quickly than among white populations. In South Carolina, one model has used primary care physicians as the labor force to provide routine screening colonoscopy for their own patients. This model makes screening much more accessible to minority patients, as the wait is shorter and the cost typically lower. In combination with a faith-based partnership with minority religious organizations, this model has begun to make needed inroads toward addressing the disparities associated with colon cancer. Cancer 2007. (c) 2006 American Cancer Society. PMID- 17123277 TI - Model for end-stage liver disease (MELD) exception small-for-size syndrome. PMID- 17123278 TI - Model for end-stage liver disease (MELD) exception for unusual metabolic liver diseases. PMID- 17123279 TI - Model for end-stage liver disease (MELD) exception for uncommon hepatic tumors. PMID- 17123280 TI - Model for end-stage liver disease (MELD) exception for bacterial cholangitis. PMID- 17123281 TI - Model for end-stage liver disease (MELD) exception for familial amyloidotic polyneuropathy. PMID- 17123282 TI - Model for end-stage liver disease (MELD) exception for hepatopulmonary syndrome. PMID- 17123283 TI - Model for end-stage liver disease (MELD) exception for portopulmonary hypertension. PMID- 17123284 TI - Model for end-stage liver disease (MELD) exception guidelines: results and recommendations from the MELD Exception Study Group and Conference (MESSAGE) for the approval of patients who need liver transplantation with diseases not considered by the standard MELD formula. PMID- 17123285 TI - Model for end-stage liver disease (MELD) exception guidelines. PMID- 17123286 TI - Model for end-stage liver disease (MELD) exception for hereditary hemorrhagic telangiectasia. PMID- 17123287 TI - Model for end-stage liver disease (MELD) exception for polycystic liver disease. PMID- 17123288 TI - Model for end-stage liver disease (MELD) exception for ascites. PMID- 17123289 TI - Model for end-stage liver disease (MELD) exception for cholangiocarcinoma or biliary dysplasia. PMID- 17123290 TI - Model for end-stage liver disease (MELD) exception for portal hypertensive gastrointestinal bleeding. PMID- 17123291 TI - Model for end-stage liver disease (MELD) exception for primary hyperoxaluria. PMID- 17123292 TI - Model for end-stage liver disease (MELD) exception for severe pruritus. PMID- 17123293 TI - Model for end-stage liver disease (MELD) exception for Budd-Chiari syndrome. PMID- 17123294 TI - Model for end-stage liver disease (MELD) exception for hepatic encephalopathy. PMID- 17123296 TI - Lactadherin detects early phosphatidylserine exposure on immortalized leukemia cells undergoing programmed cell death. AB - BACKGROUND: Phosphatidylserine (PS) appears on the outer membrane leaflet of cells undergoing programmed cell death and marks those cells for clearance by macrophages. Macrophages secrete lactadherin, a PS-binding protein, which tethers apoptotic cells to macrophage integrins. METHODS: We utilized fluorescein-labeled lactadherin together with the benchmark PS Probe, annexin V, to detect PS exposure by flow cytometry and confocal microscopy. Immortalized leukemia cells were treated with etoposide, and the kinetics and topology of PS exposure were followed over the course of apoptosis. RESULTS: Costaining etoposide-treated leukemoid cells with lactadherin and annexin V indicated progressive PS exposure with dim, intermediate, and bright staining. Confocal microscopy revealed localized plasma membrane staining, then diffuse dim staining by lactadherin prior to bright generalized staining with both proteins. Annexin V was primarily localized to internal cell bodies at early stages but stained the plasma membrane at the late stage. Calibration studies suggested a PS content less, less than or approximately equal to 2.5%-8% for the membrane domains that stained with lactadherin but not annexin V. CONCLUSIONS: Macrophages may utilize lactadherin to detect PS exposure prior to exposure of sufficient PS to bind annexin V. The methodology enables detection of PS exposure at earlier stages than established methodology. PMID- 17123297 TI - Predicting implantation with a neuromodulator using two different test stimulation techniques: A prospective randomized study in urge incontinent women. AB - AIMS: The purpose of this study was to determine whether a percutaneous needle electrode (PNE) technique or a surgical first stage lead placement (FSLP) better predicted whether a patient would progress to implantation of a pulse generator (IPG) in older urge incontinent women. METHODS: Thirty subjects > or =55 years with refractory urge incontinence who had been selected to undergo a test stimulation procedure were randomized to either PNE or FSLP. Thirteen underwent PNE placement and seventeen underwent FSLP placement. If during the test stimulation period subjects had greater than 50% improvement in their incontinence parameters they qualified for permanent lead and/or IPG implantation of the Interstim device. RESULTS: Twenty-one subjects (70%) responded to the test stimulation and underwent implantation, 15/17 (88%) in the FSLP group and 6/13 (46%) in the PNE group. Subjects who were randomized to the FSLP group were significantly more likely to proceed to implantation of the IPG (P = 0.02) than those in the PNE group. There was no significant difference in demographics, pre test stimulation incontinence parameters or post-stimulation visual analog pain scores between the randomized groups or between test stimulation responders and non-responders. When comparing FSLP and PNE responders, there was no significant difference in the percent improvement in 24-hr pad weight, daily pad usage, or daily incontinence. CONCLUSION: FSLP better predicted progression to implantation of the IPG than a test stimulation with a PNE in an older urge incontinent cohort. PMID- 17123298 TI - Smooth muscle caveolae differentially regulate specific agonist induced bladder contractions. AB - AIMS: Caveolae are cholesterol-rich plasmalemmal microdomains that serve as sites for sequestration of signaling proteins and thus may facilitate, organize, and integrate responses to extracellular stimuli. While previous studies in the bladder have demonstrated alterations in caveolae with particular physiologic or pathologic conditions, little attention has been focused on the functional significance of these organelles. Therefore, the purpose of this study was to investigate the role of caveolae in the modulation of receptor-mediated signal transduction and determine the presence and localization of caveolin proteins in bladder tissue. METHODS: Contractile responses to physiologic agonists were measured in rat bladder tissue before and after disruption of caveolae achieved by depleting membrane cholesterol with methyl-beta-cyclodextrin. Stimulation with agonists was repeated after caveolae were restored as a result of cholesterol replenishment. RT-PCR, immmunohistochemistry, and Western blotting were used to determine the expression and localization of caveolin mRNA and proteins. RESULTS: Following caveolae disruption, contractile responses to angiotensin II and serotonin were attenuated, whereas responses to bradykinin and phenylephrine were augmented. Cholesterol replenishment restored responses towards baseline. Carbachol and KCl induced contractions were not affected by caveolae disruption. Ultrastructure analysis confirmed loss of caveolae following cholesterol depletion with cyclodextrin and caveolae restoration following cholesterol replacement. Gene and protein expression of caveolin-1, -2, and -3 was detected in bladder tissue. Immunoreactivity for all three caveolins was observed in smooth muscle cells throughout the bladder. CONCLUSIONS: The functional effects of cholesterol depletion on specific agonist-induced contractile events and the expression of all three caveolins in bladder smooth muscle support a central role for caveolae in regulation of selective G-protein-coupled receptor signaling pathways in bladder smooth muscle. Thus, caveolae serve to differentially regulate bladder smooth muscle by a stimulus-dependent potentiation or inhibition of bladder contraction. PMID- 17123299 TI - M2 mediated contractions of human bladder from organ donors is associated with an increase in urothelial muscarinic receptors. AB - AIMS: Previous studies have shown increased density of M2 receptors in hypertrophied rat bladders that possess an M2 contractile phenotype. The aim of the current study is to determine whether human bladders with an M2 contractile phenotype also have a greater density of bladder M2 receptors. MATERIALS AND METHODS: Human bladders were obtained from 24 different organ transplant donors. Darifenacin and methoctramine affinity was determined by the rightward shift of cumulative carbachol concentration contractile response curves for each bladder. Radioligand binding and immunoprecipitation was used to quantify M2 and M3 subtypes in isolated detrusor muscle and urothelium. In addition, pig bladder muscle and urothelial receptors were quantified for comparison. RESULTS: In the human urothelium total, M2 and M3 muscarinic receptor density is significantly negatively correlated with the affinity of darifenacin for inhibition of contraction of the detrusor muscle. In the detrusor muscle there is no correlation between receptor density and darifenacin affinity for inhibition of contraction. Muscarinic receptor density is greater in the muscle than in the urothelium in human bladders whereas in the pig bladder the density is greater in the urothelium than in the muscle. CONCLUSIONS: The greater density of urothelial muscarinic receptors in human bladders with lower darifenacin affinity, indicative of a greater contribution of M2 receptors to the contractile response, points towards a possible role of the urothelium in controlling M2 mediated contractile phenotype. In comparison between human and pig bladders, the distribution of muscarinic receptor subtypes in the muscle and urothelium are quite different. PMID- 17123300 TI - Explorative two-locus linkage analysis suggests a multiplicative interaction between the 7q32 and 16p13 myoclonic seizures-related photosensitivity loci. AB - In traits suspected to be governed by at least two loci, linkage analysis incorporating the joint action of both loci may improve the power to detect linkage, increase the precision of estimating locus positions and provide insight into the underlying etiological mechanism. Recently, we mapped two susceptibility loci for epilepsy-related photosensitivity (or photoparoxysmal response, PPR) at regions 7q32 (PPR1) and 16p13 (PPR2) in PPR families with prominent myoclonic seizures background (MS-related PPR). To follow-up these results and evaluate interaction effects between these regions, we conducted two-locus (2L) linkage analyses using parametric and non-parametric methods. The 2L linkage was calculated under a multiplicative (MULT) epistasis model, encompassing models where each locus is necessary but not sufficient for MS-related PPR and a heterogeneity (HET) model, encompassing models in which each locus is by itself sufficient but not necessary for MS-related PPR expression. We found maximal 2L linkage under the (MULT) model, which was significantly better than the 2L linkage under the (HET) model (P = 0.001). The 2L analyses gave no increase in power to detect linkage over the single-locus analyses nor did they improve location estimates at PPR1 and PPR2, as expected under a best-fit 2L (MULT) model in an affecteds-only analysis. Our findings suggest that the genes underlying the PPR1 and PPR2 susceptibility loci may have similar functions or act in the same biochemical pathway. PMID- 17123301 TI - MCMC-based linkage analysis for complex traits on general pedigrees: multipoint analysis with a two-locus model and a polygenic component. AB - We describe a new program lm_twoqtl, part of the MORGAN package, for parametric linkage analysis with a quantitative trait locus (QTL) model having one or two QTLs and a polygenic component, which models additional familial correlation from other unlinked QTLs. The program has no restriction on number of markers or complexity of pedigrees, facilitating use of more complex models with general pedigrees. This is the first available program that can handle a model with both two QTLs and a polygenic component. Competing programs use only simpler models: one QTL, one QTL plus a polygenic component, or variance components (VC). Use of simple models when they are incorrect, as for complex traits that are influenced by multiple genes, can bias estimates of QTL location or reduce power to detect linkage. We compute the likelihood with Markov Chain Monte Carlo (MCMC) realization of segregation indicators at the hypothesized QTL locations conditional on marker data, summation over phased multilocus genotypes of founders, and peeling of the polygenic component. Simulated examples, with various sized pedigrees, show that two-QTL analysis correctly identifies the location of both QTLs, even when they are closely linked, whereas other analyses, including the VC approach, fail to identify the location of QTLs with modest contribution. Our examples illustrate the advantage of parametric linkage analysis with two QTLs, which provides higher power for linkage detection and better localization than use of simpler models. PMID- 17123302 TI - Simple methods for assessing haplotype-environment interactions in case-only and case-control studies. AB - For investigating haplotype-environment interactions in case-control studies, one can implement statistical methods based either on a retrospective likelihood (modeling the probability of haplotype and environment conditional on disease status) or a prospective likelihood (modeling the probability of disease status conditional on haplotype and environment). Retrospective approaches are generally more powerful than prospective approaches, but require an explicit model of the joint distribution of haplotype and environmental factors in the sample with the latter being particularly unattractive to specify. To resolve this issue, we propose a number of simple retrospective procedures for haplotype-environment interaction analysis that do not require explicit modeling of environmental covariates in the sample. We first consider a cases-only procedure, followed by a simple likelihood for case-control data that is proportional to the full retrospective likelihood. Finally, we consider a retrospective procedure for inference on haplotype-environment interaction effects in matched or finely stratified case-control studies. Our methods are based on the assumptions that haplotypes and environmental covariates are independent in the target population and that disease is rare. We illustrate our approaches using case-control data from the Finland-United States Investigation of Non-Insulin Dependent Diabetes Mellitus (FUSION) genetic study and simulated data. PMID- 17123303 TI - Interpretation of simultaneous linkage and family-based association tests in genome screens. AB - Linkage and association analyses have played important roles in identifying susceptibility genes for complex diseases. Linkage tests and family-based tests of association are often applied in the same data to help fine-map disease loci or validate results. This paradigm increases efficiency by making maximal use of family data sets. However, it is not intuitively clear under what conditions association and linkage tests performed in the same data set may be correlated. Understanding this relationship is important for interpreting the combined results of both tests. We used computer simulations and theoretical statements to estimate the correlation between linkage statistics (affected sib pair maximum LOD scores) and family-based association statistics (pedigree disequilibrium test (PDT) and association in the pressure of linkage (APL)) under various hypotheses. Different types of pedigrees were studied: nuclear families with affected sib pairs, extended pedigrees and incomplete pedigrees. Both simulation and theoretical results showed that when there is no linkage or no association, the linkage and association tests are not correlated. When there is linkage and association in the data, the two tests have a positive correlation. We concluded that when linkage and association tests are applied in the same data, the type I error rate of neither test will be affected and that power can be increased by applying tests conditionally. PMID- 17123304 TI - Informative-transmission disequilibrium test (i-TDT): combined linkage and association mapping that includes unaffected offspring as well as affected offspring. AB - To date, there is no test valid for the composite null hypothesis of no linkage or no association that utilizes transmission information from heterozygous parents to their unaffected offspring as well as the affected offspring from ascertained nuclear families. Since the unaffected siblings also provide information about linkage and association, we introduce a new strategy called the informative-transmission disequilibrium test (i-TDT), which uses transmission information from heterozygous parents to all of the affected and unaffected offspring in ascertained nuclear families and provides a valid chi-square test for both linkage and association. The i-TDT can be used in various study designs and can accommodate all types of independent nuclear families with at least one affected offspring. We show that the transmission/disequilibrium test (TDT) (Spielman et al. [1993] Am. J. Hum. Genet. 52:506-516) is a special case of the i TDT, if the study sample contains only case-parent trios. If the sample contains only affected and unaffected offspring without parental genotypes, the i-TDT is equivalent to the sibship disequilibrium test (SDT) (Horvath and Laird [1998] Am. J. Hum. Genet. 63:1886-1897. In addition, the test statistic of i-TDT is simple, explicit and can be implemented easily without intensive computing. Through computer simulations, we demonstrate that power of the i-TDT can be higher in many circumstances compared to a method that uses affected offspring only. Applying the i-TDT to the Framingham Heart Study data, we found that the apolipoprotein E (APOE) gene is significantly linked and associated with cross sectional measures and longitudinal changes in total cholesterol. PMID- 17123305 TI - Accumulating quantitative trait linkage evidence across multiple datasets using the posterior probability of linkage. AB - Genome scans for complex disorders are frequently inconclusive, prompting researchers to increase sample size in an effort to obtain stronger evidence. However, increasing sample size in the presence of locus heterogeneity may actually, on average, decrease the linkage signal at a true susceptibility gene. The posterior probability of linkage, or PPL, was specifically designed to address this issue in the context of categorical trait analysis, by appropriately accumulating evidence either for or against linkage as new data are added. We now formulate a quantitative trait (QT) analog, the QT-PPL, which directly measures the evidence that a QT is linked to a genetic marker or location. The new QT-PPL is based on a classical single-locus QT likelihood with the trait parameters (allele frequency, genotypic means and variances) integrated out. We show using simulations that the QT-PPL is robust to two key modeling violations (multiple trait loci and non-normality in the form of excess kurtosis), as well as being inherently ascertainment corrected, and illustrate the advantages of the QT-PPL for accumulating linkage evidence across multiple sets of data compared to other QT linkage methods. PMID- 17123306 TI - Evaluation of recurrent nodal disease after definitive radiation therapy for nasopharyngeal carcinoma: diagnostic value of fine-needle aspiration cytology and CT scan. AB - BACKGROUND: Recurrent nodal disease in patients with nasopharyngeal carcinoma (NPC) after definitive radiotherapy presents a difficult clinical problem. This cohort of patients poses a diagnostic challenge to the head and neck surgeon because evaluation of the post-irradiated neck, both clinically and radiologically, is known to be difficult, and it is not uncommon for neck dissection specimen in suspected recurrent nodal disease to contain no viable tumor cells. Currently, there is no well-accepted method for the preoperative determination of the presence of malignancy in these nodal diseases. METHODS: Over a 7-year period in a tertiary hospital, we systematically reviewed the clinical charts of 42 patients with NPC who were diagnosed with suspected recurrent nodal disease, after radical definitive radiotherapy. Fine-needle aspiration cytology (FNAC) was performed on clinically palpable nodes and results were correlated with final histopathologic results. Findings on CT scan were also correlated with final histopathologic specimens. RESULTS: The specificity and sensitivity of FNAC was 75.0% and 75.0%, respectively. The positive and negative predictive value of FNAC was 93.8% and 37.5%, respectively. CT scan had a positive predictive value of 78.6%. The negative predictive value for multilevel involvement on CT scan was 20%. CONCLUSION: Radiological imaging and FNAC are useful diagnostic modalities in assessing recurrent nodal disease in the post irradiated neck in patients with NPC. Although routine CT scan criteria for pathologic lymphadenopathy cannot be accurately applied in the post-irradiated neck, it is a useful surveillance tool in the routine follow-up of patients with post-irradiated neck with NPC. Clinicians, however, must understand their limitations when assessing these patients. The possibility of negative neck dissection must be conveyed to the patients. PMID- 17123307 TI - Down-regulation of S100A2 in lymph node metastases of head and neck cancer. AB - BACKGROUND: Our cDNA microarray analysis of squamous cell carcinoma of the head and neck (SCCHN) previously identified that S100A2 was down-regulated in highly metastatic 686LN-M3s cell lines established through in vivo selection using a metastatic xenograft mouse model. S100A2, a putative tumor suppressor, has been found to be down-regulated in several types of primary tumor as compared with the normal tissue. Only a few reports have explored its expression status and function in metastasis. METHODS: To further confirm down-regulation of S100A2 in human metastasis, we examined S100A2 expression using immunohistochemical analysis of paraffin-embedded SCCHN tissues. The samples included primary SCCHN tumors (Tu-1) and involved lymph nodes (Met-1) from the same patients, and primary tumors in node-negative patients (Tu-2). RESULTS: Most of these tumors expressed S100A2 but lymph node metastases showed a pattern of reduced staining for S100A2 compared with primary tumors. A similar expression pattern of S100A2 was also observed in several SCCHN cell lines by reverse transcription-polymerase chain reaction (RT-PCR) and immunoblotting. Particularly, S100A2 expression was lower in 686LN than Tu686 and hardly detectable in the metastatic derivatives 686LN-M3s. Further study of S100A2 promoter showed higher methylation intensity in these metastatic derivatives than in Tu686 and 686LN. CONCLUSIONS: S100A2 was down-regulated in lymph node metastasis of SCCHN, suggesting that instead of being a putative tumor suppressor, S100A2 may play a role in the metastasis of SCCHN. PMID- 17123308 TI - Quality of integrated care for patients with head and neck cancer: Development and measurement of clinical indicators. AB - BACKGROUND: To improve the quality of integrated care, we developed indicators for assessing current practice in a large reference center for head and neck oncology. METHODS: We defined a set of indicators based on integrated care literature, national evidence-based guidelines for patients with head and neck cancer, and the opinions of professionals and patients. We tested this set regarding assessment of current practice and clinimetric characteristics. RESULTS: The final set consisted of 8 integrated care indicators and 23 specific indicators for patients with head and neck cancer. Current practice assessment produced high scores for the integrated care indicators, but the specific indicators showed room for improvement. The practice test showed that 9 indicators had good applicability. CONCLUSIONS: The indicators, while based on evidence-based guidelines and the principles of integrated care, should incorporate patients' opinions and include a practice test. Our results show that the quality of integrated care for patients with head and neck cancer could be improved. PMID- 17123309 TI - Inverse correlation between serum PGE2 and T classification in head and neck cancer. AB - BACKGROUND: Prostaglandin E2 (PGE2) serum levels have been shown previously to be increased in tumor bearing mice as well as in patients with solid tumors; however, the impact on the course or stage of disease has not been shown. We hypothesized that PGE2 is strictly required for aggressive and especially early stage tumors of the head and neck to provoke invasion and angiogenesis. METHODS: We analyzed the serum PGE2 levels of 100 patients with head and neck squamous cell carcinoma of different stages before and 1 year after treatment and compared the results with the serum levels of 40 healthy donors and the secretion profile of 8 different squamous cell carcinoma cell lines. RESULTS: Our investigations showed a statistically significant inverse correlation between PGE2 serum levels and tumor stage. Furthermore, this effect has been reflected by the results of our cell culture analyses, which showed an inversely regulated PGE2 secretion into the medium during the process of proliferation. Interestingly, the serum levels of PGE2 were significantly downregulated 1 year after successful treatment. CONCLUSIONS: We conclude that PGE2 serum level as an indicator for early-stage cancer of the head and neck may function as a tumor marker during the follow-up period. PMID- 17123310 TI - Mutant p53 proteins: between loss and gain of function. AB - Cancer might result from both the aberrant activation of genes, whose physiological tuning is essential for the life of a normal cell, and the inactivation of tumor suppressor genes, whose main job is to preserve the integrity of cell genome. Among the latter, p53 is considered a key tumor suppressor gene that is inactivated mainly by missense mutations in half of human cancers. It is becoming increasingly clear that the resulting mutant p53 proteins gain oncogenic properties favoring the insurgence, the maintenance, and the spreading of malignant tumors. In this review, we mainly discuss the molecular mechanisms underlying gain of function of human tumor-derived p53 mutants, their impact on the chemoresistance and the prognosis of human tumors, with a special focus on head and neck cancers, and the perspectives of treating tumors through the manipulation of mutant p53 proteins. PMID- 17123311 TI - Microlaryngoscopic repair of iatrogenic pharyngeal pouch perforations: treatment of choice? AB - BACKGROUND: Endoscopic stapled diverticulostomy (ESD) has become the preferred technique for managing pharyngeal pouches. Iatrogenic perforation, created during stapling, is a rare but serious complication with significant morbidity and mortality. The conventional management in these instances is to convert it to an external procedure and excise the pouch. METHODS: Iatrogenic perforations were noticed after stapling in 3 cases in our series of 73 patients who underwent ESD. They were repaired using microlaryngoscopic techniques. RESULTS: All patients had an unremarkable postoperative course. CONCLUSIONS: Selected cases with iatrogenic perforations can be repaired primarily and observed with excellent outcome, obviating the need for an external pouch excision. PMID- 17123312 TI - Cordectomy by CO2 laser or radiotherapy for small T1a glottic carcinomas: costs, local control, survival, quality of life, and voice quality. AB - BACKGROUND: The clinical results of radiotherapy and endoscopic cordectomy for T1a glottic carcinoma are reported to be similar, but costs of both treatments may differ. Therefore, we retrospectively evaluated the costs, voice quality, quality of life, and clinical results of both treatments. METHODS: Costs and effects from the first visit up to 2 years of follow-up were calculated, based on chart data of 35 and 54 patients who were treated with curative intent for T1a glottic carcinoma from 1995 to 1999 with radiotherapy and endoscopic cordectomy, respectively. Voice quality and quality of life were evaluated by means of patient questionnaires. RESULTS: Total costs (in Euros) of radiotherapy and CO(2) laser cordectomy were 8322 euro and 4434 euro, respectively, including the costs of treating possible recurrences. The effects of both treatments were equal, including voice quality and quality of life. CONCLUSIONS: We conclude that endoscopic cordectomy is an efficient alternative for radiotherapy for these patients. PMID- 17123313 TI - Actin deficiency induces cofilin phosphorylation: proteome analysis of HeLa cells after beta-actin gene silencing. AB - Actin-binding proteins regulate the dynamic structure and function of actin filaments in the cell. Much is known about how manipulation of the actin-binding proteins affects the structure and function of actin filaments; however, little is known about how manipulation of actin in the cell affects actin-binding proteins. We addressed this question by utilizing two technologies: RNA interference and 2-dimensional gel electrophoresis. We knocked down beta-actin expression in HeLa cells using short interfering RNA and applied 2-DGE to examine alterations in the HeLa cell proteome. We revealed a 2-5 fold increases of four protein spots on 2-D gels and identified these proteins by mass spectrometry. Three of the four proteins were actin-binding proteins, including cofilin, which promotes both disassembly and assembly of actin filaments but becomes inactivated when phosphorylated. Further examination revealed that the cofilin total protein level barely increased, but the phosphorylated cofilin level increased dramatically in HeLa cells after beta-actin siRNA treatment. These results suggest that in response to siRNA-induced beta-actin deficiency HeLa cells inactivate cofilin by phosphorylation rather than down-regulate its protein expression level. This study also demonstrates that the combination of RNA interference and 2-dimensional gel electrophoresis technologies provides a valuable method to study protein interactions in a specific cellular pathway. PMID- 17123314 TI - Journey in reverse: TTP from bedside to blood bank to bench. AB - Thrombotic thrombocytopenic purpura (TTP) is the most extensive and dangerous intravascular platelet clumping disorder. For more than a half-century after its initial recognition, mortality was near 100% and the etiology totally obscure. Then, in the late 1970s to early 1980s, empiric, but successful, therapy by a few clinician/blood bank partnerships was followed by sudden laboratory insight into pathophysiology. The discussion that follows was prepared in conjunction with the 2006 Francis Morrison, M.D., Memorial Lecture at the 27th Annual Meeting of the American Society for Apheresis. PMID- 17123315 TI - Prospectively assessed incidence, severity, and determinants of respiratory symptoms in the first year of life. AB - Respiratory symptoms are common in infancy. Nevertheless, few prospective birth cohort studies have studied the epidemiology of respiratory symptoms in normal infants. The aim of this study was to prospectively obtain reliable data on incidence, severity, and determinants of common respiratory symptoms (including cough and wheeze) in normal infants and to determine factors associated with these symptoms. In a prospective population-based birth cohort, we assessed respiratory symptoms during the first year of life by weekly phone calls to the mothers. Poisson regression was used to examine the association between symptoms and various risk factors. In the first year of life, respiratory symptoms occurred in 181/195 infants (93%), more severe symptoms in 89 (46%). The average infant had respiratory symptoms for 4 weeks and 90% had symptoms for less than 12 weeks (range 0 to 23). Male sex, higher birth weight, maternal asthma, having older siblings and nursery care were associated with more, maternal hay fever with fewer respiratory symptoms. The association with prenatal maternal smoking decreased with time since birth. This study provides reliable data on the frequency of cough and wheeze during the first year of life in healthy infants; this may help in the interpretation of published hospital and community-based studies. The apparently reduced risk in children of mothers with hayfever but no asthma, and the decreasing effect of prenatal smoke exposure over time illustrate the complexity of respiratory pathology in the first year of life. PMID- 17123316 TI - Human metapneumovirus and respiratory syncytial virus infections in older children with cystic fibrosis. AB - BACKGROUND: Human metapneumovirus (hMPV) has been isolated from children with acute respiratory infection worldwide. Its epidemiology remains to be defined in children with cystic fibrosis (CF). We describe the epidemiology and clinical impact of hMPV in CF children and compared it to respiratory syncytial virus (RSV). METHODS: CF children ages 7-18 years were studied prospectively during the 1998 -1999 RSV season. Nasopharyngeal specimens were collected during acute respiratory illnesses and tested for respiratory viruses. Blood specimens were drawn early, mid, and end of the RSV season, and tested for serological evidence of hMPV and RSV infections. Rates of lower respiratory tract illnesses (LRTI) and hospitalizations for pulmonary exacerbations were compared during the time intervals they developed serological evidence of infection to their non-infection intervals. RESULTS: Six of 44 CF children had a virus positive respiratory illness in 56 LTRI events and 18 hospitalizations. Serological evidence of hMPV and RSV infections occurred in 16 and 20 CF children, respectively; 8 had infections with both viruses. A greater proportion of CF children had >or=1 LRTI during their infection intervals compared to their non-infection intervals (13/25 vs. 5/25; P=0.03). A trend for higher rates of LRTI was observed in the infection intervals compared to non-infection intervals (9.5 +/- 11.0 vs. 4.2 +/- 9.9 per 1,000 child-days; P=0.06), and it was significantly greater with a more conservative estimate (one event per child per interval; 7.4 +/- 7.7 vs. 2.6 +/- 5.4 per 1,000 child-days; P or=25mU/L in every patient in whom levels were measured (n = 112). Hypothyroid complications were avoided in 22 of 25 patients who had experienced them in the past, and in 47 of 51 patients who were at high risk for hypothyroid complications. Radioiodine uptake was present on whole-body scans (WBS) in 105 of the 115 patients. Serum thyroglobulin levels were lower than baseline in 73% of patients assessed at 12 months. Cancer related symptoms were improved in approximately 25%. Two patients had serious adverse events that were thought to be related to rhTSH. rhTSH elevates serum TSH and facilitates radioiodine uptake in patients who cannot produce endogenous TSH or who cannot tolerate hypothyroidism. PMID- 17123340 TI - Toward the application of proteomics to human thyroid tissue. AB - CONTEXT: In this paper we describe for the first time a systematic approach to proteome analysis of human thyroid tissue. OBJECTIVE AND DESIGN: We report different methods to decrease the complexity of the human thyroid tissue proteome by applying different solubilization strategies and correcting for thyroglobulin protein abundance; to increase the protein resolution by prefractionation and by the use of narrow-range pH gradients; to detect proteins using sensitive and quantitative stains; and to identify soluble and membrane-bound thyroid tissue proteins by mass spectrometry analysis. MAIN OUTCOME/RESULTS: We found that buffers containing high contents of urea and detergents allow the best solubilization of human thyroid tissue proteins; highly variable abundance of thyroglobulin is a major pitfall of human thyroid proteome analysis, which in contrast to centrifugal ultrafiltration, size-exclusion chromatography and microdissection, can be countered best by adapting the protein amount to the thyroglobulin content per sample; prefractionation leads to a significant enrichment of proteins and allows subcellar localization of thyroid proteins; application of narrow-range immobilized pH gradient (IPG) strips allows further improvement of spot detection and separation; and protein detection with the fluorescent stain ruthenium II Tris bathophenanthroline disulfonate (RuBPs) is a highly sensitive and reliable tool for quantitative proteome analysis. Finally, in a pilot study of four patients with benign nodular thyroid disease we found that the described procedures allow a highly reproducible detection and identification of alterations in protein expression between nodular and corresponding normal thyroid tissues. CONCLUSIONS: Application of the described methods provides the basis for a highly sensitive and reproducible proteome analysis of the human thyroid, providing an additional novel tool to elucidate complex proteins changes in human thyroid biology as well as pathophysiology of human thyroid disease. PMID- 17123341 TI - Positive predictive value of detectable stimulated tg during the first year after therapy of thyroid cancer and the value of comparison with Tg-ablation and Tg measured after 24 months. AB - This study evaluated the positive predictive value (PPV) of detectable stimulated thyroglobulin during the first year after treatment of thyroid carcinoma (Tg-1) and the value of comparison with Tg-ablation and measured after 24 months (Tg-2). Forty-two consecutive patients undergoing total thyroidectomy and ablation with detectable Tg-1 (>1ng/mL) were selected. The patients had well-differentiated tumors, which were completely resected, and there was no ectopic uptake on whole body scan after 3.7-5.5GBq I(131). Imaging methods during follow-up revealed metastases in 10 patients (24%) (15% if Tg-1 10 ng=mL). Tg-ablation (cutoff of 10 ng/mL) presented a negative predictive value (NPV) of 91% and PPV of 42%. Comparing Tg-ablation with Tg-1, the PPV of an increase was 100%, whereas the NPV of a decrease was 88%. Thirty-six patients presented negative imaging results upon first assessment and Tg-1 was compared to Tg-2. Metastases were detected in all patients who presented an increase in Tg (n=4), whereas patients without variation (n=4) or with a decrease (n=28) showed no apparent disease. Among disease-free patients (n=32), 50% presented undetectable Tg and 40% showed a >50% decrease after 2 years. In conclusion, most patients with detectable stimulated Tg during the first year after therapy had no metastases, and evaluation of the slope of Tg helped discriminate cases with apparent disease. PMID- 17123342 TI - Trend in thyroid carcinoma size, age at diagnosis, and histology in a retrospective study of 500 cases diagnosed over 20 years. AB - Recently, the Italian Network of Cancer Registries analyzed 5101 cases of thyroid carcinoma showing a reduction of mortality rate of 4%/year. This prompts us to evaluate the temporal trend in tumor size, age at diagnosis, and histology in a retrospective analysis of 500 thyroid cancers diagnosed over 20 years. Thyroid cancers were divided in two groups. The first included 193 cases diagnosed from 1985 to 1994, and the second 307 from 1995 to 2004. The size of all tumors was significantly reduced from 30 +/- 1.4mm in the first group to 15 +/- 0.8mm in the second group. In particular, papillary thyroid carcinoma (PTC) size decreased from 28 +/- 1.2mm to 14 +/- 0.8mm and follicular carcinoma from 40 +/- 6.3mm to 17 +/- 4.5 mm. Age at diagnosis of all carcinomas increased significantly from 40 +/- 1.3 years in the first group to 48 +/- 0.9 years in the second group. Analysis of the histological types revealed a significant increase of PTC rate in the second decade from 82% to 92% and a concomitant reduction of anaplastic thyroid carcinoma (ATC) from 3.7% to 1.0%. Moreover, a significant increase of micro-PTC rate, from 7.3% to 36.4%, was observed. In conclusion, it may be speculated that the above mentioned decreased mortality rate for thyroid carcinoma could be related to the significant reduction with time of cancer size, to the progressive increase of PTC rate and to the reduction of ATC rate. These data, if confirmed in other series, underscore the importance of evaluating thyroid nodules smaller than 10mm and corroborate recent findings suggesting that age be reconsidered as an independent prognostic factor for differentiated thyroid cancers. PMID- 17123343 TI - Action of topical thyroid hormone analogue, triiodothyroacetic acid in reversing glucocorticoid-induced skin atrophy in humans. AB - The present study concerns the effect of topical treatment with a cream formulation of triiodothyroacetic acid (TRIAC) in comparison with a placebo preparation in producing a reversal of skin atrophy induced by long-term employment of topical glucocorticoid therapy in humans. A total of 39 patients with clinically verified skin atrophy due to long-term use of topical potent glucocorticoids were randomized. The changes in skin thickness, elastic fibers, and hyaluronic acid were evaluated by means of sonography and histology. After 8 weeks' treatment, the skin thickness measured by sonography increased by 16% in the epidermis, 8% in the dermis, and epidermis + dermis in the placebo group. In the TRIAC 0.1% group, the corresponding values were 24% ( p=0.063) in the epidermis, 28% ( p=0.042) in the dermis, and 25% ( p=0.039) in the epidermis + dermis. After 8 weeks, in the placebo group, the skin thickness measured by biopsy increased by 5% in the epidermis, epidermis + dermis, and 6% in the dermis. In the TRIAC 0.1% group, the corresponding values were 31% ( p=0.041) in the epidermis, 46% ( p=0.041) in the dermis and 44% ( p=0.043) in the epidermis + dermis. After 8 weeks, the elastic fibers of moderately irregular and thickened fibers increased by 56% in the placebo group and 100% ( p=0.043) in the TRIAC 0.1 group. This study indicates that topical treatment with TRIAC appears to reverse glucocorticoid-induced skin atrophy under the narrow conditions tested. PMID- 17123344 TI - Biochemical markers in the follow-up of medullary thyroid cancer. AB - Medullary thyroid cancer (MTC) shares biochemical features with other neuroendocrine tumors but the particular characteristics are largely unexplored. We investigated the biochemical neuroendocrine profile of MTC and whether specific markers could be useful in follow-up. In addition to the standard tumor marker calcitonin, plasma carcino-embryonic antigen (CEA), plasma catecholamines, (platelet) serotonin, chromogranin A, tryptase, and urinary markers of catecholamine, histamine, and serotonin metabolism were prospectively determined in 46 patients with histologically proven MTC. Patients were divided according to the stage of disease: group 1, no evidence; group 2, stable disease (SD); and group 3, progressive disease (PD). Plasma dopamine was increased in the majority of the patients with SD and PD; however it did not correlate with extent of disease. Elevated plasma platelet levels of serotonin were only present in patients with multiple endocrine neoplasia 2 with SD or PD but did not differ between those groups. Histamine metabolites were elevated in 20% of patients with SD and PD. In addition to plasma calcitonin, only CEA and chromogranin A could differentiate between stable and progressive MTC. MTCs are capable of synthesizing catecholamines, serotonin, and histamine metabolites underscoring that MTCs have metabolic characteristics in common with other neuroendocrine tumors. Thus far, clinical usefulness and relevance seems limited. The most useful markers in the follow-up of MTC are plasma calcitonin and CEA. PMID- 17123345 TI - Lactose intolerance revealed by severe resistance to treatment with levothyroxine. AB - The most common cause of apparent ineffectiveness or resistance to treatment with oral levothyroxine (LT(4)) is the result of noncompliance, known as pseudomalabsorption. However, an abnormality in the bioavailability of LT(4) should also be considered in patients requiring large doses of LT(4) to achieve euthyroidism. The incidence of lactose intolerance in Caucasian adult patients is 7%-20%, but the association with resistance to treatment with oral LT(4) is unusual. We report a 55-year-old woman in whom treatment LT(4) for hypothyroidism was found related to a previously undiagnosed oligo-symptomatic lactose intolerance, an unusual association. Although rare, intolerance to lactose should be considered in the differential diagnosis of gastrointestinal diseases that can cause malabsorption of LT(4). The possibility of correcting this disorder with simple dietary measures justifies its consideration. PMID- 17123346 TI - Thyroid sarcoidosis as a unique localization. AB - Sarcoidosis is a systemic disorder characterized by granulomatous lesions, principally affecting the lungs. There are numerous reports in the literature of an associated involvement of the thyroid, much more frequently in hypothyroid than in hyperthyroid subjects. The present case report refers to a woman presenting with thyroid nodules and normal biochemical levels and thyroid function parameters, while histology revealed sarcoid-type lesions. Subsequent investigations and a long follow-up showed no evidence of involvement of other sites, including the lungs. Consequently, after a long period of normal health associated with the negative results of the examinations, a diagnosis of sarcoidosis limited to the thyroid was proposed, which was cured by thyroidectomy. PMID- 17123347 TI - Ectopic intrathyroidal thymus in an 11-year-old boy. PMID- 17123348 TI - Mitochondrial abnormalities in eye muscle fiber from three cases of thyroid associated ophthalmopathy. PMID- 17123349 TI - Exercise-induced radio-iodine accumulation in scalp and hair during admission of 131I therapy for thyroid cancer. PMID- 17123350 TI - Artifactual I-131 activity in the hairs after thyroid remnant ablation. PMID- 17123351 TI - Preventive medicine: a "cure" for the healthcare crisis. PMID- 17123352 TI - Identification and expression of a sialyltransferase responsible for the synthesis of disialylgalactosylgloboside in normal and malignant kidney cells: downregulation of ST6GalNAc VI in renal cancers. AB - Although disialyl glycosphingolipids such as GD3 and GD2 have been considered to be associated with malignant tumours, whether branched-type disialyl glycosphingolipids show such an association is not well understood. We investigated the sialyltransferases responsible for the biosynthesis of DSGG (disialylgalactosylgloboside) from MSGG (monosialylgalactosylgloboside). Among six GalNAc:alpha2,6-sialyltransferases cloned to date, we focused on ST6GalNAc III, V and VI, which utilize sialylglycolipids as substrates. In vitro enzyme analyses revealed that ST6GalNAc III and VI generated DSGG from MSGG with V(max)/K(m) values of 1.91 and 4.16 respectively. Transfection of the cDNA expression vectors for these enzymes resulted in DSGG expression in a renal cancer cell line. Although both ST6GalNAc III and VI genes were expressed in normal kidney cells, the expression profiles of ST6GalNAc VI among 20 renal cancer cell lines correlated clearly with those of DSGG, suggesting that the sialyltransferase involved in the synthesis of DSGG in the kidney is ST6GalNAc VI. ST6GalNAc-VI and DSGG were found in proximal tubule epithelial cells in normal kidney tissues, while they were downregulated in renal cancer cell lines and cancer tissues. All these findings indicated that DSGG was suppressed during the malignant transformation of the proximal tubules as a maturation arrest of glycosylation. PMID- 17123353 TI - SARP, a new alternatively spliced protein phosphatase 1 and DNA interacting protein. AB - PP1 (protein phosphatase 1) is a ubiquitously expressed serine/threonine-specific protein phosphatase whose activity towards different substrates appears to be mediated via binding to specific proteins that play critical regulatory and targeting roles. In the present paper we report the cloning and characterization of a new protein, termed SARP (several ankyrin repeat protein), which is shown to interact with all isoforms of PP1 by a variety of techniques. A region encompassing a consensus PP1-binding motif in SARP (K354VHF357) modulates endogenous SARP-PP1 activity in mammalian cells. This SARP-PP1 interaction motif lies partially within the first ankyrin repeat in contrast with other proteins [53BP2 (p53 binding protein 2), MYPT1/M(110)/MBS (myosin binding protein of PP1) and TIMAP (transforming growth factor beta inhibited, membrane-associated protein)], where a PP1-binding motif precedes the ankyrin repeats. Alternative mRNA splicing produces several isoforms of SARP from a single human gene at locus 11q14. SARP1 and/or SARP2 (92-95 kDa) are ubiquitously expressed in all tissues with high levels in testis and sperm, where they are shown to interact with both PP1gamma1 and PP1gamma2. SARP3 (65 kDa) is most abundant in brain where SARP isoforms interact with both PP1alpha and PP1gamma1. SARP is highly abundant in the nucleus of mammalian cells, consistent with the putative nuclear localization signal at the N-terminus. The presence of a leucine zipper near the C-terminus of SARP1 and SARP2, and the binding of mammalian DNA to SARP2, suggests that SARP1 and SARP2 may be transcription factors or DNA-associated proteins that modulate gene expression. PMID- 17123354 TI - p56lck, LFA-1 and PI3K but not SHP-2 interact with GM1- or GM3-enriched microdomains in a CD4-p56lck association-dependent manner. AB - We previously showed that the association of CD4 and G(M3) ganglioside induced by CD4 ligand binding was required for the down-regulation of adhesion and that aggregation of ganglioside-enriched domains was accompanied by transient co localization of LFA-1 (lymphocyte function-associated antigen-1), PI3K (phosphoinositide 3-kinase) and CD4. We also showed that these proteins co localized with the G(M1) ganglioside that partially co-localized with G(M3) in these domains. In the present study, we show that CD4-p56(lck) association in CD4 signalling is required for the redistribution of p56(lck), PI3K and LFA-1 in ganglioside-enriched domains, since ganglioside aggregation and recruitment of these proteins were not observed in a T-cell line (A201) expressing the mutant form of CD4 that does not bind p56(lck). In addition, we show that although these proteins associated in different ways with G(M1) and G(M3), all of the associations were dependent on CD4-p56(lck) association. Gangliosides could associate with these proteins that differ in affinity binding and could be modified following CD4 signalling. Our results suggest that through these associations, gangliosides transiently sequestrate these proteins and consequently inhibit LFA-1-dependent adhesion. Furthermore, while structural diversity of gangliosides may allow association with distinct proteins, we show that the tyrosine phosphatase SHP-2 (Src homology 2 domain-containing protein tyrosine phosphatase 2), also required for the down-regulation of LFA-1-dependent adhesion, transiently and partially co-localized with PI3K and p56(lck) in detergent-insoluble membranes without association with G(M1) or G(M3). We propose that CD4 ligation and binding with p56(lck) and their interaction with G(M3) and/or G(M1) gangliosides induce recruitment of distinct proteins important for CD4 signalling to form a multimolecular signalling complex. PMID- 17123387 TI - Inhibitor treatment in haemophilias A and B: summary statement for the 2006 international consensus conference. AB - Participants in an international conference on the management of haemophilia patients with inhibitors developed a jointly authored summary of the findings and conclusions of the conference. Current knowledge of the genetic and immunologic mechanisms underlying inhibitor development was briefly summarized. Concerning the purported treatment-related risk factors, conference participants commented on the limitations of the available evidence and the need for more rigorous prospective research in a fully genotyped population. Other clinical considerations discussed included the unproved utility of routine surveillance, the need for assay standardization, the management of acute bleeding and approaches to joint disease prophylaxis and immune tolerance induction (ITI). A number of issues were identified as needing further investigation in larger prospective studies, ideally through international cooperation. Such studies should enroll cohorts that have been scrupulously defined in terms of mutation status and treatment exposure. Finally, conference participants urged their colleagues to participate in the currently ongoing international trials of ITI. PMID- 17123388 TI - Basic aspects of inhibitors to factors VIII and IX and the influence of non genetic risk factors. AB - The appearance of polyclonal antibodies inhibiting the function of exogenous factors VIII (FVIII) and IX (FIX) continues to be a major challenge in the treatment of patients with congenital haemophilia. Why these inhibitors develop in 10-20% of patients with haemophilia A, and in 1-5% of patients with haemophilia B, remains largely unexplained. The antibodies, however, are characterized by several features that may have implications for the immune process by which they occur. The FVIII antibodies are mainly directed towards the A2, A3 and C2 domains, thereby interfering with the function of the factor Xase complex, the binding of FVIII to von Willebrand factor, and the binding of FVIII to phospholipid membranes. The FIX epitopes are localized to the NH(2)-terminal gamma-carboxyglutamic acid region and the serine protease domain. Genetic risk factors are known to be of importance in the development of inhibitors, whereas the impact of non-genetic factors is less clear. However, based on studies of related subjects, it is obvious that non-genetic factors are of importance as well. Putative factors currently debated include age at the start of treatment, treatment in association with immune challenges, the type of product, and the mode of administration. Most of the findings reported to date, however, derive from small cohorts that have not been sufficiently well characterized with respect to genetic risk profile. Therefore, additional studies are required to quantify the impact of non-genetic factors on the pathophysiologic process of inhibitor development. PMID- 17123389 TI - Genetic risk factors for inhibitors to factors VIII and IX. AB - The formation of alloantibodies against factor VIII (FVIII) or factor IX (FIX) is the most severe complication of replacement therapy in patients with haemophilia. In the last decade, genetic factors have been shown to constitute a decisive risk determinant for the development of inhibitors. In severe haemophilia A and B, mutations that result in an absent or truncated FVIII/FIX protein are associated with a 20-80% risk of inhibitor formation. In mild to moderate haemophilia, missense mutations represent the main mutation type, with an inhibitor prevalence of 5%. These patients synthesize some endogenous, although non-functional protein that is sufficient to induce immune tolerance. However, in patients with missense mutations clustered in the A2 and C2 domains (C1/C2 junction), the risk of inhibitor formation is fourfold greater than in patients with mutations outside this region, indicating that inhibitor prevalence in missense mutations is also dependent on localization of the mutation. Recently, a significant association between inhibitor formation and polymorphisms in genes coding for cytokines (IL 10) and other immunoregulatory factors (TNF-alpha) has been shown. These genetic factors constitute the individual genetic risk profile of a haemophilic patient. This risk is imprinted and fixed; however, environmental factors such as treatment schedule may increase or decrease the inhibitor risk in an individual patient. Improved understanding of these complex interactions may lead to the development of preventive measures to minimize inhibitor formation. PMID- 17123390 TI - The epidemiology of factor VIII inhibitors. AB - Risk factors for inhibitor development include specific factor VIII (FVIII) genotypes, a family history of inhibitor development amongst first-degree relatives, certain HLA haplotypes and non-caucasian ethnicity. Patients with major FVIII deletions or rearrangements have a higher risk of inhibitor development than those with small deletions or missense mutations. Studies of HLA DR amongst northern European patients with the intron 22 inversion indicate that certain HLA haplotypes may also confer either increased or decreased inhibitor risk. However, although brothers of patients with inhibitor have a high risk of inhibitor development, concordance is not 100%, and other constitutional factors must also operate. Disputed risk factors for inhibitor development include early age at first FVIII exposure, the use of recombinant rather than plasma-derived FVIII and product switching. Two studies suggest that inhibitor risk is increased by very early FVIII exposure (P = 0.03), but this is disputed by a third study; larger studies addressing the question are ongoing. Studies investigating the relative inhibitor risk of plasma-derived vs. recombinant FVIII have been similarly inconclusive and will be reviewed. Other environmental or treatment variables such as concomitant infection or vaccination are suspected to influence inhibitor risk but have not yet been fully investigated. PMID- 17123391 TI - Immunological aspects of inhibitor development. AB - The development of inhibitor antibodies is perhaps the most serious complication of coagulation factor replacement therapy. A complex interaction of several variables leads to inhibitor formation in congenital haemophilia, while acquired haemophilia represents a failure of the immune tolerance mechanisms that regulate a normal immune response to factor VIII (FVIII). The immune response to FVIII is dependent upon the interaction of different CD4+ T-cell subsets (Th1, Th2 and Th3) specific for FVIII. Failure to activate regulatory CD4+ cells likely plays a crucial role in the development of FVIII inhibitors. Although the basic mechanisms of the immune response to FVIII in the setting of factor replacement therapy are being elucidated, a clear understanding of the relevance of these mechanisms in the context of successful immune tolerance therapy and ultimately gene therapy, awaits further study. PMID- 17123392 TI - Inhibitor treatment in haemophilias A and B: inhibitor diagnosis. AB - The clinical diagnosis and quantitative measurement of polyclonal IgG inhibiting antibodies are the subjects of this review. Inhibitors in congenital haemophilia are usually diagnosed either as part of a routine surveillance schedule or following a bleeding episode that responds poorly to standard specific replacement therapy. Routine surveillance schedules for paediatric haemophilia A patients during high-risk incidence periods are variable and the subject of ongoing discussion. There have never been any published recommendations for following haemophilia B patients at high risk for inhibitor development. The Factor VIII/IX Subcommittee of the International Society on Thrombosis and Haemostasis scientifically endorsed the Nijmegen method for inhibitor measurement in 1996. However, there are many unresolved issues surrounding inhibitor diagnosis using these assays. These issues include: (i) questions of accuracy and inter-assay variability inherent to the one-stage clotting assay; (ii) lack of consensus regarding the assay cut-off for negative determination; (iii) lack of assay standardization and (iv) the clinical importance of capturing non neutralizing antibodies currently not measured in the functional assays. Ongoing efforts to resolve these issues will be discussed. PMID- 17123393 TI - Characteristics of inhibitors in mild/moderate haemophilia A. AB - Patients with mild or moderate haemophilia A usually have a mild bleeding disorder requiring only occasional treatment with factor VIII (FVIII) concentrates. The frequency of inhibitor development in such patients has been the subject of several recent surveys, which significantly modified our appreciation of this complication. Studies of the anti-FVIII antibodies provided an explanation for the different bleeding phenotypes observed in mild/moderate haemophilia A patients with inhibitors. Antibodies distinguishing between the patient's mutant FVIII and the normal wild-type FVIII were characterized, in addition to antibodies inhibiting completely or only partially FVIII activity. T lymphocytes recognizing FVIII and likely involved in the development of the immune response to FVIII were successfully identified. The FVIII peptides recognized by those FVIII-specific cells bind to many major histocompatibility complex (MHC) class II molecules, which may provide an explanation for the lack of strong association between MHC haplotypes and inhibitor development. Although these studies have advanced our understanding of the conditions leading to inhibitor development, further work is required to determine whether the mode of FVIII administration significantly influences inhibitor development. Further studies of the genetic factors are also required to fully understand the mechanisms leading to inhibitor development in patients with mild/moderate haemophilia A. PMID- 17123394 TI - Basic aspects of bypassing agents. AB - Bypassing agents consist of activated prothrombin complex concentrates (aPCC) and recombinant factor VIIa (rFVIIa). Their main utilization is for prevention and treatment of bleeding complications, which may occur in inhibitor-developing haemophiliacs, although new indications for rFVIIa (e.g. trauma-related and cerebral bleeds) are now under evaluation in clinical trials. The mechanisms of action for these agents are still not fully understood. The relative complexity of the composition of aPCC suggests the possibility of multiple modes of action for achieving haemostasis. Among those possibilities, the contributions of activated factor X and prothrombin have been demonstrated in recent years both in vitro and in animal models for the only aPCC which remains on the market. rFVIIa also exhibits a complex mode of action, improving coagulation through both tissue factor-dependent and -independent pathways. The various mechanisms that occur at the cellular surfaces, particularly on the outer leaflet of the platelet membrane, primarily contribute to Xase complex formation and thrombin generation. The ways in which these agents affect the complex kinetics of fibrin formation at the site of vascular damage need further clarification, although significant progress has been achieved in the last 10 years. In addition, the ex vivo monitoring that would reflect achievement of haemostasis in vivo is still not standardized, although several attempts using thromboelastography, thrombin generation and the kinetics of fibrin formation have been initiated. PMID- 17123395 TI - An overview of the history, clinical practice concerns, comparative studies and strategies to optimize therapy of bypassing agents. AB - Despite significant advances in the treatment of haemophilia, including availability of recombinant coagulation factor replacement products and the use of prophylactic infusion regimens, the segment of haemophilic patients who develop inhibitory antibodies remain at higher risk for morbidity and mortality associated with recurrent or uncontrolled bleeding events. Bypassing agents represent the mainstay of treatment and prevention of bleeding. The most commonly used of the currently available therapeutic agents are a plasma-based therapy, factor eight inhibitor bypassing activity, vapour heated, and a recombinant therapy, NovoSeven (recombinant activated factor VIIa). A substantial body of literature exists to document efficacy and adverse event profiles for these two products. There is, however, a paucity of data arising from adequately powered prospective trials to determine optimal treatment and dosing in various clinical situations. Certain clinical circumstances, patient profiles, individual responses, or provider predilection may lead to preferential use of one of these products over the other; however, the continued presence of both agents in the therapeutic armamentarium remains critically important to this fragile population. The historical use, clinical practice concerns, published comparative studies and methods for optimization of these two bypassing agents are reviewed. PMID- 17123396 TI - Options for treating acute bleeds in addition to bypassing agents: extracorporeal immunoadsorption, FVIII/FIX, desmopressin and antifibrinolytics. AB - Inhibitor patients do not always respond satisfactorily to treatment with bypassing agents, and options to the standard practice are sometimes needed. Temporary inhibitor removal may be achieved using extracorporeal immunoadsorption. This technique uses a column system including either protein A or antihuman IgG. Immunoadsorption may be used as part of an immune tolerance protocol, or in the case of acute bleeds or prior to surgery, thus rendering the patient more responsive to ordinary replacement therapy with factor VIII or factor IX. Desmopressin is a valuable haemostatic agent in many situations and can be especially recommended in mild haemophilia complicated by an inhibitor. Antifibrinolytics are often administered as an adjunct therapy to the treatment protocol and have also been reported to have a direct anti-inhibitor effect. PMID- 17123397 TI - Prophylaxis in haemophilia patients with inhibitors. AB - The presence of high titre inhibitors makes the treatment of bleeding episodes in haemophilia patients difficult and increases the risk of uncontrollable bleeding and disability, despite optimum on-demand treatment with bypassing agents. The inability to effectively control joint bleeding leads to progressive joint disease in many patients with inhibitors. Significant mobility impairments are far more prevalent in patients with inhibitors than in those without inhibitors. Emerging data suggest that prophylaxis using bypassing agents may be effective and safe in reducing the incidence of joint bleeding during immune tolerance induction (ITI), and for patients who failed ITI or who were never candidates for ITI. Only controlled clinical trials will ultimately demonstrate whether prophylaxis can prevent joint bleeding and damage, and improve quality of life in patients with inhibitors. This article will review the published data on the use of bypassing agents in the prevention of bleeding, and will discuss ongoing clinical prophylaxis trials in inhibitor patients. PMID- 17123398 TI - Assessing the costs for clinical care of patients with high-responding factor VIII and IX inhibitors. AB - The costs of haemophilia-related care and the impact of unusually expensive, or outlier, patients on these costs have been explored in numerous European, American and Canadian studies during the last decade. In particular, antibodies that neutralize infused factor VIII or IX (high-responding inhibitors) make treatment responses, and thereby costs, much less predictable. There is little debate that the health care costs of haemophilic patients with high-responding inhibitors are routinely higher and more variable than those of non-inhibitor patients. However, the extent to which this is attributable to the few outlier inhibitor patients whose expenditures tend to skew the data is not as clear. To compare the variation and range in health care expenditures among patients with inhibitors and those without, we reviewed data originally gathered during a 24 month period in 1995-1997 from a prospectively created cohort as part of a broader cost and utilization study conducted at a large haemophilia treatment centre. We conclude that although the use of outpatient factor replacement products was not significantly greater or more expensive among inhibitor patients, their hospital-related costs greatly increased overall expenditures. Among our study population, the overall costs associated with inhibitor patients are not only higher in absolute monetary terms, but also in terms of the degree of variation. This variation was demonstrated by: (i) the extremely wide range of costs over an extended timeframe among individual inhibitor patients when compared with those without inhibitors, and (ii) the much larger year-to-year variation in costs among the inhibitor group. PMID- 17123399 TI - Immune tolerance: critical issues of factor dose, purity and treatment complications. AB - The current practice of immune tolerance induction (ITI) therapy has been largely influenced by the results of small institutional studies and three large registries. However, many questions remain. Successful outcome predictors for ITI in haemophilia A have been suggested by the analyses of two of these registries. Among these predictors, factor VIII (FVIII) dose/dosing regimen remains a controversial outcome parameter, demonstrating a strong direct relationship to ITI success in the international registry and a weaker inverse relationship in the North American registry. There is an international multicentre prospective randomized trial underway to further study the role of FVIII dose in successful ITI induction in a good risk haemophilia A inhibitor patient cohort. FVIII purity also remains an unproved ITI outcome predictor. Institutional experience with von Willebrand-factor-containing products has suggested its therapeutic advantage in both inhibitor development and eradication. The International ITI Study, although not designed to answer this particular question, may be able to determine an impact on outcome depending on the final distribution of investigator choice of product among the study subjects. Much less is known about the influence of factor IX (FIX) dose and purity on ITI success in haemophilia B. Importantly, nephrotic syndrome has been a major determinant of ITI failure in FIX inhibitor patients, particularly those with the allergic phenotype. Unfortunately, large prospective randomized trials in this group will not be feasible. Rather, we will have to rely on prospectively collected registry data to build our knowledge base of inhibitors and ITI in haemophilia B. PMID- 17123400 TI - Overview of the use of implantable venous access devices in the management of children with inherited bleeding disorders. AB - Frequent infusion of factor concentrates may be challenging in young boys with haemophilia, especially if their disease is complicated by inhibitors. A central venous access device (CVAD) is often placed in young patients in need of repeated infusions for prophylaxis or immune tolerance induction. Although user friendly and capable of providing reliable venous access, these devices are associated with a high complication rate over time. In the haemophilia population, major complications include CVAD-associated infections and deep venous thrombosis, which is most often silent. Established risk factors for catheter-related infection include age less than 6 years at the time of CVAD placement and use of an external CVAD when compared with a totally implantable device such as a port. Avoidance of CVAD-related infections is facilitated by strict adherence to aseptic technique. The risk of deep venous thrombosis appears related to the duration for which the catheter is in place, with the risk increasing beyond 4 years. The promotion of a strict clinic policy in which CVADs are left in place for as short a time as possible should decrease the risk of complications. In rare cases where a totally implantable CVAD cannot be placed for technical reasons, an arteriovenous fistula may provide reliable venous access. In all cases, however, venous access via peripheral veins is preferred over CVADs. PMID- 17123401 TI - Novel therapies for immune tolerance in haemophilia A. AB - Standard treatment for an inhibitory antibody to factor VIII (FVIII) in a patient with severe congenital haemophilia A is to attempt to induce immune tolerance with high-dose FVIII, either alone or in combination with immunosuppression. Patients in whom the inhibitor is not eradicated or who have prognostic features suggestive of a poor response may be suitable for experimental approaches to immune tolerance induction. The two options that are currently under investigation in clinical practice are anti-CD20 antibody therapy using rituximab and the use of von Willebrand factor (VWF)-containing FVIII concentrate in immune tolerance regimens. Immunomodulation with rituximab has been reported to eradicate inhibitors in some patients with severe haemophilia A who have previously failed standard immune tolerance. Similarly, some patients who have failed to be tolerized with high-purity FVIII have been successfully treated with VWF-containing concentrates. Neither of these treatment modalities is supported by controlled clinical trial data, and reported observational data require confirmation. Immunomodulation via interference with B-cell/T-cell interactions by blocking CD40/CD40 ligand or a gene therapy approach using FVIII peptides in IgG heavy chain transfected into B-cell blasts has been reported to suppress inhibitors in animal models and may lead to clinically useful therapies. Further understanding of the aetiology of inhibitor formation and how FVIII leads to tolerance in some patients with an inhibitor may suggest further approaches in the future. PMID- 17123402 TI - Recombinant vs. plasma-derived products, especially those with intact VWF, regarding inhibitor development. AB - The development of inhibitors in previously untreated patients (PUPs) with haemophilia A is correlated with a variety of endogenous and exogenous risk factors. It is still controversial whether recombinant factor VIII (rFVIII) products pose a higher risk for inhibitor development than plasma-derived (pd) FVIII concentrates, particularly with intact von Willebrand factor (VWF). A systematic review on the epidemiology of inhibitors in haemophilia A investigated the influence of different FVIII products on inhibitor formation. Patients treated with a single pd product had a lower cumulative incidence (0-12.4%) than those treated with a single recombinant concentrate (36.0-38.7%) independent of disease severity, study size, or inhibitor testing frequency. However, this analysis does not take into account the heterogeneity of the study populations. A study investigating the effects of different variables on inhibitor development in PUPs was started 13 years ago by the Paediatric Committee of the German, Austrian and Swiss Society on Thrombosis and Haemostasis Research. This prospectively conducted study revealed a slight difference (P = 0.08) in terms of type of concentrate. In the group of severe haemophiliacs treated with pdFVIII concentrate (n = 57), 12 patients developed an inhibitor (21%), whereas 17 of 47 patients receiving rFVIII (36%) were affected by this complication. This observation was substantiated by retrospective Italian and French analyses. These results, as well as hypothetical considerations, indicate that there is a body of evidence that FVIII products with VWF may be less immunogenic in PUPs. However, in order to provide more evidence a well-designed randomized study is needed. PMID- 17123403 TI - Urgent inhibitor issues: targets for expanded research. AB - Although up to 80% of high-responding inhibitors in patients with severe factor VIII deficiency can be eliminated using heterogeneous regimens for immune tolerance induction, the residual morbidity in this population of haemophilic patients is far from trivial. There is an exigent need for focussed basic, translational and clinical research to extend our understanding of the pathogenesis of haemophilic inhibitor development. In this article, we identify four key research needs, including (i) whether presently available clotting factor concentrates (CFCs) have differential antigenicity, giving rise to clinically relevant immunogenicity; (ii) the interplay of quantitative and qualitative (e.g. age at first exposure) influences of CFCs as well as host environmental factors (e.g. vaccination effects) on inhibitor development; (iii) the therapeutic role (if any) that concurrent immune tolerance with suppressive or immune-competitive therapeutic strategies play in inhibitor eradication and (iv) pending any major therapeutic advances, alternative or enhanced strategies for treating acute haemorrhage and for preventing chronic haemorrhagic events in these patients. PMID- 17123404 TI - Two new species of symbiotic ciliates from the respiratory tract of cetaceans with establishment of the new genus Planilamina n. gen. (Dysteriida, Kyaroikeidae). AB - Examination of mucus discharged from the blowhole of Atlantic bottlenose dolphin (Tursiops truncatus) at Marine Life Oceanarium, Gulfport, Mississippi, and false killer whale (Pseudorca crassidens) and Atlantic bottlenose dolphin at SeaWorld Orlando, Orlando, Florida, using live observations and protargol impregnation revealed mixed infections of Kyaroikeus cetarius and two new species. Planilamina n. gen. is characterized by a C-shaped argentophilic band located along the laterally flattened margin of cell and extending from the cell apex to subposterior cone-shaped podite; a deep oral cavity containing one short preoral kinety, two circumoral kineties, seven to 13 infundibular kineties, and a cytostome; a broadly funnel-shaped cytopharynx reinforced by argentophilic fibers but without nematodesmata; closely packed postoral kinetofragments set in a pocket located anterior left of the podite; and somatic kineties as a right field closely situated at the right surface and a left field bordering the anterior left margin of the oral cavity. The type species for the genus, Planilamina ovata n. sp., is distinguished from its sister species Planilamina magna n. sp. by the following characteristics: body size (28-65 x 20-43 microm vs. 57-90 x 40-63 microm), number of right field kineties (38-55 vs. 79-99), and position of the anterior end of the leftmost kinety in the right somatic field (anterior one third vs. mid-body). The morphogenesis of Planilamina ovata is similar to that of K. cetarius. The diagnosis of Kyaroikeidae is emended to accommodate the new genus. PMID- 17123405 TI - Protist genetic diversity in the acidic hydrothermal environments of Lassen Volcanic National Park, USA. AB - We examined eukaryote genetic diversity in the hydrothermal environments of Lassen Volcanic National Park (LVNP), Northern California. We sampled hydrothermal areas of the Bumpass Hell, Sulfur Works, Devil's Kitchen, and Boiling Springs Lake sites, all of which included diverse acidic pools, mud pots, and streams with visible algal mats and biofilms. Temperatures varied from 15 to 85 degrees C and pH from 1.7 to 5.8. DNA extraction methods compared by denaturing gradient gel electrophoresis fingerprinting exhibited similar patterns, and showed limited diversity of eukaryotic small subunit (SSU) rRNA genes compared with prokaryotes. We successfully amplified eukaryotic SSU rRNA genes from most environments up to 68 degrees C. Cloned rDNA sequences reveal acidophilic protists dominate eukaryotes in LVNP hydrothermal environments. Most sites showed phototrophic assemblages dominated by chlorophytes and stramenopiles (diatoms and chrysophytes). Heterotrophic taxa, though less abundant, included diverse alveolates (ciliates), amoebae, and flagellates. Fungi were also found at most sites, and metazoans (hexapods, nematodes, platyhelminths) were sometimes detected in less acidic environments, especially in algal mats. While many cloned rDNA sequences showed 95%-99% identity to known acidophilic isolates or environmental clones from other acidic sites (Rio Tinto), sequence diversity generally declined both with decreasing pH and increasing temperature, and both were controlling physical variables on the abundance and distribution of organisms at our sites. However, a pool at 68 degrees C with pH 1.7 yielded the greatest number of distinct sequences. While some were likely contaminants from nearby cooler sites, we suggest that Lassen's acidic hydrothermal features may harbor novel protists. PMID- 17123406 TI - Artemia is capable of spreading oocysts of Cryptosporidium and the cysts of Giardia. AB - The capability for ingesting and spreading the fixed oocysts of Cryptosporidium and fixed cysts of Giardia, two waterborne protozoan parasites, by Artemia franciscana, a microcrustacean widely used as live diet in fish and shellfish larviculture, was demonstrated using differential interference contrast and immunofluorescence microscopy. Our findings suggest the possibility that this microcrustacean could serve as a disseminating vehicle of both parasites in aquatic environments. PMID- 17123407 TI - Ubiquinone synthesis and its regulation in Pneumocystis carinii. AB - The opportunistic pathogen Pneumocystis causes a type of pneumonia in individuals with defective immune systems such as AIDS patients. Atovaquone, an analog of ubiquinone (coenzyme Q [CoQ]), is effective in clearing mild to moderate cases of the infection. Rat-derived Pneumocystis carinii was the first organism in which CoQ synthesis was clearly demonstrated to occur in both mitochondrial and microsomal subcellular fractions. Atovaquone inhibits microsomal CoQ synthesis with no effect on mitochondrial CoQ synthesis. We here report on additional studies evaluating CoQ synthesis and its regulation in the organism. Buparvaquone also inhibited CoQ synthesis and it reduced the synthesis of all four CoQ homologs in the microsomal but not the mitochondrial fraction. Glyphosate, which inhibits a reaction in the de novo synthesis of the benzoquinone moiety of CoQ reduced cellular ATP levels. Bacterial and plant quinones, and several chemically synthesized phenolics, flavanoids, and naphthoquinones that inhibit electron transport in other organisms were shown to reduce CoQ synthesis in P. carinii. The inhibitory action of naphthoquinone compounds appeared to depend on their molecular size and structural flexibility rather than redox potential. Results of experiments examining the synthesis of the polyprenyl chain of CoQ were consistent with negative feedback control of CoQ synthesis. These studies on P. carinii suggest that cellular sites and the control of CoQ synthesis in different organisms and cell types might be more diverse than previously thought. PMID- 17123408 TI - Phylogenetic distance of Thelohania butleri Johnston, Vernick, and Sprague, 1978 (Microsporidia; Thelohaniidae), a parasite of the smooth pink shrimp Pandalus jordani, from its congeners suggests need for major revision of the genus Thelohania Henneguy, 1892. AB - Thelohania butleri, a microsporidian that causes mortality and commercial losses in the smooth pink shrimp Pandalus jordani, is of taxonomic interest as a species resembling the poorly studied type species, Thelohania giardi, of the large, polyphyletic genus Thelohania. We examined the ultrastructure of T. butleri to confirm its identity and reconstructed phylogenies using ribosomal DNA to find the relationship of T. butleri with other Thelohania species in crayfish and ants. Light and transmission electron microscopy from specimens collected from the type locality, the Pacific coast of Canada, confirmed the identity and demonstrated a development similar to that of T. giardi, involving a series of binary fissions without formation of a plasmodium. Phylogenetic analyses consistently showed T. butleri to be distantly related to other Thelohania species, and closely related to species from marine decapods within a larger fish parasitic clade. Together, features such as host group and habitat, developmental morphology, and phylogeny suggest T. butleri may be a closer relative to T. giardi than any other Thelohania species represented by DNA data so far, and thus imply species from crayfish and ants may not belong in this genus. Results also confirm that genus Thelohania and family Thelohanidae are in need of revision. PMID- 17123409 TI - Cytopathogenicity of Balamuthia mandrillaris, an opportunistic causative agent of granulomatous amebic encephalitis. AB - Balamuthia mandrillaris is a free-living ameba and an opportunistic agent of lethal granulomatous amebic encephalitis in humans and other mammals. Balamuthia mandrillaris is highly cytopathic but, in contrast to the related Acanthamoeba, does not feed on bacteria and seems to feed only on eukaryotic cells instead. Most likely, the cytopathogenicity of B. mandrillaris is inseparable from its infectivity and pathogenicity. To better understand the mechanisms of B. mandrillaris cytopathogenicity, an assay for measuring amebic cytolytic activity was adapted that is based on the release of a reporter enzyme by damaged target cells. The ameba is shown to lyse murine mastocytoma cells very efficiently in a time- and dose-related manner. Furthermore, experiments involving semipermeable membranes and phagocytosis inhibitors indicate that the cytolytic activity of B. mandrillaris is essentially cell contact-dependent. Standard and fluorescence light microscopy, as well as scanning and transmission electron microscopy support and extend these findings at the ultrastructural level. PMID- 17123410 TI - Trachipleistophora extenrec n. sp. a new microsporidian (fungi: microsporidia) infecting mammals. AB - A new microsporidian Trachipleistophora extenrec n. sp. was isolated from a muscle lesion of the streaked tenrec Hemicentetes semispinosus Cuvier, 1798 (Mammalia, Tenrecidae), an insectivore endemic to Madagascar. The spores isolated from the tenrec were infectious to severe combined immunodeficient (SCID) mice by intramuscular injection. Material obtained from muscular lesions in mice was used for the parasite description. All developmental stages of the microsporidian were covered by a dense coat, which during sporogony changed into the sporophorous vesicle wall. Eight, 16, 32, or more spores were formed inside the sporophorous vesicle as the result of the division by plasmotomy and sequential fission of a multinucleate sporogonial plasmodium. Spores were ovoid, 4.7 x 2.8 microm in size, had a large posterior vacuole, and had an isofilar polar tube with 15-16 coils. Although the fine structure and the developmental pattern of the organism were in some respects similar to the genus Vavraia, molecular phylogeny based on the gene sequences of the small subunit rRNA and RNA polymerase subunit II indicated that the organism belongs to the genus Trachipleistophora. The diagnostic characters of the genera Trachipleistophora and Vavraia are discussed as well as the discrepancies between the phylogenies of these two microsporidian genera based on morphology and molecules. PMID- 17123411 TI - Pelagostrobilidium wilberti n. sp. (Oligotrichea, Choreotrichida): morphology and morphogenesis. AB - Morphology, infraciliature, morphogenetic features, and some ecological data for Pelagostrobilidium wilberti n. sp. are described. This new species was collected from a temporary pond in Magdalena, Buenos Aires province, Argentina, which was sampled monthly from August 2003 to July 2005. The species was found in autumn and winter. Observations were made in vivo and after staining with protargol. Pelagostrobilidium wilberti n. sp. measures 63-84 x 42-49 microm in vivo and is conical in shape, with a posterior spine-like cytoplasmic process. It possesses 6 somatic kineties, with kinety 2 sinistrally curved and shorter than the others. The oral apparatus is composed of 25-32 external and two internal membranelles. The macronucleus is horseshoe-shaped and located beneath the oral apparatus; two or three spherical micronuclei lie dorsally. There is a posterior contractile vacuole. Morphogenesis is hypo-apokinetal and begins dorsally between the curved kinety 2 and kinety 3. After the discovery of this new species, the diagnosis of the genus Pelagostrobilidium was amended. PMID- 17123412 TI - Tunicothrix rostrata n. g., n. sp., a new urostylid ciliate (Ciliophora, Stichotrichia) from the Yellow Sea. AB - The morphology and infraciliature of a new ciliate, Tunicothrix rostrata n. g., n. sp., isolated from the Yellow Sea, are investigated using live observation and protargol impregnation. Tunicothrix rostrata measures about 160 x 40 mum in vivo, and has a frontal beak-like protrusion, a conspicuous cortical alveolar layer, two right marginal rows, and usually three distinct midventral pairs. The discovery of T. rostrata enables us to reconsider the classification of Erniella wilberti, a curious ciliate with obscure midventral pairs. Both species are highly similar in overall appearance and nuclear and ciliary pattern. Thus, they are united in a new genus of the family Urostylidae, Tunicothrix, and E. wilberti is transferred to Tunicothrix: Tunicothrix wilbertiLin and Song, 2004 n. comb. By contrast, Erniella filiformis, type species of Erniella, has several ventral rows and does not belong to the urostylids. Tunicothrix rostrata is easily distinguished from T. wilberti by its beak-like anterior protrusion and by the distinctly elongated right marginal row 2, which curves anteriorly on the dorsal side of the cells. Tunicothrix is closely related to Parabirojimia, differing by the invariably two (vs. five-eight) right marginal rows and the conspicuous (vs. ordinary) alveolar layer, a unique feature in urostylid ciliates. PMID- 17123413 TI - A molecular phylogenetic investigation of zoothamnium (ciliophora, peritrichia, sessilida). AB - The gene coding for 18S small subunit ribosomal RNA (ssu rRNA) was sequenced in seven free-living, marine species of the sessiline peritrich genus Zoothamnium. These were Zoothamnium niveum, Zoothamnium alternans, Zoothamnium pelagicum, and four unidentified species. The ssu rRNA gene also was sequenced in Vorticella convallaria, Vorticella microstoma, and in an unidentified, freshwater species of Vorticella. Phylogenetic trees were constructed using these new sequences to test a previously published phylogenetic association between Zoothamnium arbuscula, currently in the family Zoothamniidae, and peritrichs in the family Vorticellidae. Trees constructed by means of neighbor-joining, maximum parsimony, maximum likelihood, and Bayesian inference methods all had similar topologies. The seven new sequences of Zoothamnium species grouped into three well-supported clades, each of which contained a diversity of morphological types. The three clades formed a poorly supported, larger clade that was deeply divergent from Z. arbuscula, which remained more closely associated with vorticellid peritrichs. It is apparent that Zoothamnium is a richly diverse genus and that a much more intensive investigation, involving both morphological and molecular data and a wider selection of species, will be necessary to resolve its phylogeny. A greater amount of molecular diversity than is predicted by morphological data exists within all major clades of sessiline peritrichs that have been included in molecular phylogenies, indicating that characteristics of stalk and peristomial structure traditionally used to differentiate taxa at the generic level and above may not be uniformly reliable. PMID- 17123414 TI - Molecular characterization of the obligate endosymbiont "Caedibacter macronucleorum"Fokin and Gortz, 1993 and of its host Paramecium duboscqui strain Ku4-8. AB - Bacterial endosymbionts of protozoa were often described as new species by protozoologists mainly on the basis of few morphological characters and partly by host specificity. Many of these species have never been validated by prokaryotic microbiologists whose taxonomic rules are quite different from those of protozoologists, who use the Zoological Code of Nomenclature. "Caedibacter macronucleorum"Fokin and Gortz 1993, an endosymbiont of Paramecium duboscqui, belongs to this category. Here we provide the molecular characterization of this organism and of its host P. duboscqui strain Ku4-8. Bacterial 16S rRNA gene sequence analysis proved that "C. macronucleorum" belongs to the Alphaproteobacteria. It is closely related to Caedibacter caryophilus but not to Caedibacter taeniospiralis, which belongs to the Gammaproteobacteria. "Caedibacter macronucleorum" and C. caryophilus 16S rRNA genes show a similarity value of 99%. This high 16S rRNA sequence similarity and the lack of a specific oligonucleotide probe for distinguishing the two endosymbionts do not allow validating "C. macronucleorum" as a provisional taxon (Candidatus). Nevertheless, "C. macronucleorum" and C. caryophilus can be easily discriminated on the basis of a highly variable stretch of nucleotides that interrupts the 16S rRNA genes of both organisms. PMID- 17123415 TI - Effects of altered temperature and precipitation on desert protozoa associated with biological soil crusts. AB - Biological soil crusts are diverse assemblages of bacteria, cyanobacteria, algae, fungi, lichens, and mosses that cover much of arid land soils. The objective of this study was to quantify protozoa associated with biological soil crusts and test the response of protozoa to increased temperature and precipitation as is predicted by some global climate models. Protozoa were more abundant when associated with cyanobacteria/lichen crusts than with cyanobacteria crusts alone. Amoebae, flagellates, and ciliates originating from the Colorado Plateau desert (cool desert, primarily winter precipitation) declined 50-, 10-, and 100-fold, respectively, when moved in field mesocosms to the Chihuahuan Desert (hot desert, primarily summer rain). However, this was not observed in protozoa collected from the Chihuahuan Desert and moved to the Sonoran desert (hot desert, also summer rain, but warmer than Chihuahuan Desert). Protozoa in culture began to encyst at 37 degrees C. Cysts survived the upper end of daily temperatures (37-55 degrees C), and could be stimulated to excyst if temperatures were reduced to 15 degrees C or lower. Results from this study suggest that cool desert protozoa are influenced negatively by increased summer precipitation during excessive summer temperatures, and that desert protozoa may be adapted to a specific desert's temperature and precipitation regime. PMID- 17123416 TI - Novel nucleomorph genome architecture in the cryptomonad genus hemiselmis. AB - Cryptomonads are ubiquitous aquatic unicellular eukaryotes that acquired photosynthesis through the uptake and retention of a red algal endosymbiont. The nuclear genome of the red alga persists in a highly reduced form termed a nucleomorph. The nucleomorph genome of the model cryptomonad Guillardia theta has been completely sequenced and is a mere 551 kilobases (kb) in size, spread over three chromosomes. The presence of three chromosomes appears to be a universal characteristic of nucleomorph genomes in cryptomonad algae as well as in the chlorarachniophytes, an unrelated algal lineage with a nucleomorph and plastid genome derived from a green algal endosymbiont. Another feature of nucleomorph genomes in all cryptomonads and chlorarachniophytes examined thus far is the presence of subtelomeric ribosomal DNA (rDNA) repeats at the ends of each chromosome. Here we describe the first exception to this canonical nucleomorph genome architecture in the cryptomonad Hemiselmis rufescens CCMP644. Using pulsed field gel electrophoresis (PFGE), we estimate the size of the H. rufescens nucleomorph genome to be approximately 580 kb, slightly larger than the G. theta genome. Unlike the situation in G. theta and all other known cryptomonads, sub telomeric repeats of the rDNA cistron appear to be absent on both ends of the second largest chromosome in H. rufescens and two other members of this genus. Southern hybridizations using a variety of nucleomorph protein gene probes against PFGE-separated H. rufescens chromosomes indicate that recombination has been a major factor in shaping the karyotype and genomic structure of cryptomonad nucleomorphs. PMID- 17123417 TI - Vannella epipetala n. sp. isolated from the leaf surface of Spondias mombin (Anacardiaceae) growing in the dry forest of Costa Rica. AB - As part of a Microbial Observatory of Caterpillars located in the Area de Conservacion Guanacaste (ACG) in northwestern Costa Rica, we isolated a novel species of the genus Vannella associated with the food of the caterpillars of the saturniid moth Rothschildia lebeau, namely the leaves of the dry forest deciduous tree Spondias mombin (Anacardiaceae). The new species can be distinguished from other described species of the genus by the presence of a plasmalemma coated with a thickened, osmiophilic lamina containing glycostyles, and by its unusual habitat, the leaf surfaces or phylosphere of S. mombin. We further established the novelty of our isolate by sequencing its nuclear small-subunit (SSU) rRNA gene and inferring its phylogenetic position among all other currently sequenced members of the genera Vannella and Platyamoeba. Our results reveal that our isolate shares most recent common ancestry with three strains of Platyamoeba placida, the type species of the genus Platyamoeba. Despite this placement, the isolate clearly possesses glycostyles that are the hallmark of the genus Vannella. In addition to the cultured isolate, we also present a closely related sequence from a SSU rRNA gene clone library constructed from a DNA extract of leaf-wash of S. mombin with sterile water. PMID- 17123418 TI - The marine pathogenic genotype of Spironucleus barkhanus from farmed salmonids redescribed as Spironucleus salmonicida n. sp. AB - There are two genotypes of the diplomonad Spironucleus barkhanus. Based on sequence data from the small subunit ribosomal RNA gene the conspecificity of these two genotypes has been questioned. Therefore, we have sampled Spironucleus from 27 fish, representing 14 populations, five species, and four genera. Partial nucleotide sequences from the three genes; small subunit ribosomal DNA, glutamate dehydrogenase 1 and alpha-tubulin were compared. The pathogenic isolates of S. barkhanus, which causes systemic spironucleosis in Atlantic salmon, Chinook salmon, and Arctic charr, all farmed in sea water, were genetically very different from the commensal isolate found in wild freshwater populations of Arctic charr and grayling. The genetic distances between the genotypes were of the same magnitude as those separating species of Giardia. Based on these genetic and ecological data, the pathogenic genotype from farmed salmonids is described as a new species, Spironucleus salmonicida n. sp. Scanning and transmission electron microscopy showed no specific morphological or ultrastructural features distinguishing S. salmonicida n. sp. from S. barkhanus. The present study clearly demonstrates the value of applying genetics in identification of Spironucleus species. Phylogenetic analyses that included the isolates of S. salmonicida n. sp. did not change the phylogenetic relationship within the genus Spironucleus. PMID- 17123422 TI - Protection of gruntlings against classical swine fever virus-infection after oral vaccination of sows with C-strain vaccine. AB - The objective of this study was to investigate the maternal protection of gruntlings derived from wild sows vaccinated orally against classical swine fever (CSF) using C-strain vaccine. Three vaccinated sows and one unvaccinated control sow were included. Challenge infection of the progeny was carried out either intranasally or by contact at the beginning of the third month of life (61-65 days post-natum). Whereas, two of three litters had maternal antibodies, the progeny of one vaccinated sow was seronegative at challenge. The progeny of the control sow, which was challenged by contact infection, developed moderate clinical signs except for one animal which became ill and died. Two gruntlings derived from the vaccinated sows also died of CSF, although one of them had a relatively high maternal antibody titre (128 ND(50)). The transient infection and partial virus shedding observed in a small number of gruntlings with maternal antibodies and the fact that one animal with maternal antibodies became ill and died confirm the incomplete maternal protection at this age. The reason for this incomplete protection is discussed. As none of the surviving gruntlings could be shown to carry CSFV or viral RNA at the end of the experiment (36 or 70 d.p.i.), it may be concluded that these animals do not represent a potential CSFV reservoir. PMID- 17123423 TI - First results of detection of PRRSV and CSFV RNA by SYBR Green I-based quantitative PCR. AB - Porcine reproductive and respiratory syndrome (PRRS) and classical swine fever (CSF) cause significant economic losses to the swine industry worldwide. As both diseases cause similar symptoms, rapid and reliable detection of these diseases is essential for disease surveillance. A quantitative SYBR Green I-based reverse transcription-polymerase chain reaction (RT-PCR) is described for simultaneous and differential diagnosis. The established RT-PCR for the quantitation of PRRSV and CSFV cDNA was found to provide a broad dynamic range, detecting from 10(3) to 10(11) and 10(2) to 10(11) copies of cDNA per reaction, respectively. Sensitivity and specificity of this method were compared with those of conventional RT-PCR and both were equal or superior to the reference method. Reproducibility was tested and the assay was proved very reliable. The assay is timesaving, easy to handle, and highly sensitive and specific. Therefore, it is a powerful tool for detecting PRRSV and CSFV simultaneously for routine outbreak investigation. PMID- 17123424 TI - First detection of canine parvovirus type 2c in pups with haemorrhagic enteritis in Spain. AB - Canine parvovirus type 2 (CPV-2), the aetiological agent of haemorrhagic enteritis in dogs, includes three antigenic variants, types 2a, 2b and 2c. CPV-2c has been detected initially in Italy and subsequently in Vietnam. We report the first identification of this novel antigenic variant in Spain, where it caused an outbreak of fatal enteritis in basset hound pups in association with canine coronavirus type I and type II. We suggest that this new antigenic variant of CPV 2 could spread throughout Europe and that there is a subsequent need to update current CPV vaccines. PMID- 17123425 TI - A simple statistical approach for assessing inter-laboratory comparison tests for classical swine fever diagnosis. AB - The aim of this study was to compare on an objective basis the results obtained during five classical swine fever (CSF) ring tests conducted in Germany between 1999 and 2003. A novel and simple statistical approach used in behavioural sciences was used. For each ring test, the regional laboratories received a panel of five lyophilized pig sera. The panel contained CSF virus positive and negative samples. The final task of the laboratory was to ascertain if a serum sample was positive for CSF or not. Some sera were very easy to diagnose as CSF positive while some sera had border line values and proved to be challenging. Depending on the degree of difficulty the sera were divided into five categories. The evaluation of the ring test results was performed using a scoring system based on a score from -3 to +3 which takes into consideration the degree of difficulty to produce a correct diagnosis. To compare the results between different laboratories and/or between different ring tests more easily the total score of one laboratory was expressed in percentage. The final analysis of the data showed that the CSF diagnostic quality improved continuously. PMID- 17123426 TI - Serological, bacteriological and molecularbiological survey of paratuberculosis (Johne's disease) in Austrian cattle. AB - Paratuberculosis (Johne's disease) is a chronic infectious disease of ruminants, caused by Mycobacterium avium subspecies paratuberculosis (MAP). Because of its long incubation period, high economic losses, difficulties in diagnosis and possible links to Morbus Crohn in humans, paratuberculosis is one of the most important diseases of ruminants today. An abattoir-based nationwide survey on the occurrence of MAP in the Austrian cattle population was performed using serology (SVANOVIR-ELISA) as well as culture, ZN-stain and IS900-PCR on faeces and lymph node samples. A total of 756 Austrian slaughter cattle were serologically, bacteriologically and molecularbiologically tested for the occurrence of MAP and specific antibodies respectively. Samples were collected following a statistical plan to obtain balanced specimens from the whole country. Nineteen per cent of the animals tested were serological positive, 10.1% gave an inconclusive result and 70.9% showed no specific antibodies against MAP. Only in four individuals MAP could be detected by stain, bacteriology or Polymerase Chain Reaction. The calculated prevalence of 19.0% positive cattle, each representing one farm, showing specific antibodies against MAP is rather high in terms of animal-level but low in herd level prevalence compared with other countries. When this study is compared with a similar study performed in Austria 1999, a significant increase of positive cattle and farms could be seen in Austria. PMID- 17123427 TI - Canine isosporosis - epidemiology of field and experimental infections. AB - Isospora spp. are the causative agents of canine isosporosis. Of the 3590 diagnostic samples from Austrian dogs (< or = 2 years old), 8.7% contained Isospora oocysts, 78% of which from dogs up to 4 months of age. Non-haemorrhagic and haemorrhagic diarrhoea were significantly more prevalent in Isospora-infected animals than in coccidia-negative ones. Twelve of 15 litters from a large commercial dog breeding unit (examined from the third to the 10th week of life) also excreted Isospora (average prevalence: 36.4%) in intensities from 333 to 35,000 oocysts per gram of faeces (opg). In experimental trials 26 3-week-old Beagle puppies were infected with low (600-6000), medium (10,000) or high (20,000) dose of Isospora ohioensis-group or Isospora canis field isolates. Additionally 21 puppies were infected as above and treated with a symmetrical triazintrione. Parasitological and clinical parameters were examined. The two Isospora species differed significantly concerning intensity and duration of excretion. The pre-patent period was 6-7 days for I. ohioensis and 10-12 days for I. canis. The latter species showed significantly longer excretion and higher opg. This was not influenced by simultaneous infections with both species. Individual patterns of faecal consistency were very variable, irrespective of the infection dose. Treatment significantly reduced both the intensity and the duration of oocysts excretion as well as diarrhoea in comparison with the infected, untreated group and thus proved to be effective against coccidiosis in experimental infections. PMID- 17123428 TI - Porcine acute phase protein concentrations in different diseases in field conditions. AB - Five acute phase proteins (APPs) [C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp), pig-MAP and albumin] were measured in pigs with naturally occurring infections by porcine reproductive and respiratory syndrome virus (PRRSV), Aujeszky's disease virus (ADV), porcine circovirus type 2 (PCV2) and Mycoplasma hyopneumoniae, and in animals with tail and ear bites, arthritis and other acute inflammatory processes. Healthy specific pathogen-free (SPF) pigs were used as controls. In PRRSV-infected pigs, all APPs with the exception of pig MAP exhibited significant changes compared with controls. In animals affected with ADV only Hp presented changes of statistical significance, whereas pigs with PCV2 showed marked modifications in all APPs tested. Animals affected with Mycoplasmosis showed concentrations of all positive APPs significantly above levels obtained in SPF pigs, though albumin concentrations did not differ from controls. Finally, all APPs studied showed substantial changes in pigs with acute inflammation. The results indicated that an acute phase response was developed in the different diseases studied, this response being higher in animals with clinical signs and concurrent bacterial processes. Haptoglobin would be the APP that better reflects pathological states; however, to get more complete and valuable information it might be advisable to perform APPs profiles including another APP, such as CRP or SAA. PMID- 17123429 TI - Development and validation of a competitive ELISA kit for the serological diagnosis of ovine, caprine and bovine brucellosis. AB - A competitive ELISA (Brucella-Ab c-ELISA) was standardized and validated for the detection of Brucella antibodies in cattle, sheep and goat sera using a monoclonal antibody (MAb 4B5A) produced against Brucella melitensis biotype 2. The specificity and sensitivity of the assay were 100% to a 67.5% cut-off point (B/Bo%). When compared with an indirect ELISA, the Brucella-Ab c-ELISA did not demonstrate cross-reactions when testing positive sera for antibodies to some Enterobacteriaceae. A comparison was made between the Brucella-Ab c-ELISA and the complement fixation and Rose Bengal tests. Results demonstrated that the Brucella Ab c-ELISA is a valuable tool for the serological diagnosis of bovine and ovine/caprine brucellosis. PMID- 17123430 TI - Effects of machine-milking on the bacterial flora of teat duct and mammary gland of ewes. AB - In total, 308 paired-samples of teat duct material and milk, were collected before and 50-70 min after machine-milking, from 30 ewes. Samples were processed bacteriologically. For analysis of results, we compared changes in bacterial isolation following milking, for duct and milk samples; statistical significance was assessed by the Sign Test. Bacteria were isolated from 18 (6%) duct and 19 (6%) milk samples collected before the milking procedure; respective figures after it, were 81 (26%) and 33 (11%). In 77 (25%) cases, bacteriological findings in the two duct samples of each pair were different; in seven cases bacteria were isolated only before, whilst in 70 cases bacteria were isolated only after milking (P < 0.005); respective results for milk samples were 26 (8%): 6 and 20 cases (P = 0.693). The majority of bacterial isolates were staphylococci, accounting for 63% of 99 isolates. The milking procedure predisposes to entrance of bacteria into the teat duct; however, increased bacterial isolation from the teat did not result to increased mammary infections, likely as a consequence of defence mechanisms present in healthy teats. PMID- 17123432 TI - Iatrogenesis, inflammation and organ injury: insights from a murine model. AB - The complex biology of critical illness not only reflects the initial insult that brought the patient to the intensive care unit but also, and perhaps even more importantly, it reflects the consequences of the many clinical interventions initiated to support life during a time of lethal organ system insufficiency. The latter may amplify or modify the response to the former and are eminently amenable to modulation by changes in practice. However, they rarely figure in conceptual models of critical illness and are almost never accounted for in preclinical models of disease. In the preceding issue of Critical Care, O'Mahony and colleagues reported on an animal model in which sequential insults--low-dose endotoxin followed by mechanical ventilation--induce much greater remote organ injury than either insult alone. Although animal models are poor surrogates for clinical illness, studies such as these provide valuable platforms for probing the complex interactions between insult and therapy that give rise to the intricate biology of critical illness. PMID- 17123434 TI - Systematic assignment of thermodynamic constraints in metabolic network models. AB - BACKGROUND: The availability of genome sequences for many organisms enabled the reconstruction of several genome-scale metabolic network models. Currently, significant efforts are put into the automated reconstruction of such models. For this, several computational tools have been developed that particularly assist in identifying and compiling the organism-specific lists of metabolic reactions. In contrast, the last step of the model reconstruction process, which is the definition of the thermodynamic constraints in terms of reaction directionalities, still needs to be done manually. No computational method exists that allows for an automated and systematic assignment of reaction directions in genome-scale models. RESULTS: We present an algorithm that - based on thermodynamics, network topology and heuristic rules - automatically assigns reaction directions in metabolic models such that the reaction network is thermodynamically feasible with respect to the production of energy equivalents. It first exploits all available experimentally derived Gibbs energies of formation to identify irreversible reactions. As these thermodynamic data are not available for all metabolites, in a next step, further reaction directions are assigned on the basis of network topology considerations and thermodynamics-based heuristic rules. Briefly, the algorithm identifies reaction subsets from the metabolic network that are able to convert low-energy co-substrates into their high-energy counterparts and thus net produce energy. Our algorithm aims at disabling such thermodynamically infeasible cyclic operation of reaction subnetworks by assigning reaction directions based on a set of thermodynamics derived heuristic rules. We demonstrate our algorithm on a genome-scale metabolic model of E. coli. The introduced systematic direction assignment yielded 130 irreversible reactions (out of 920 total reactions), which corresponds to about 70% of all irreversible reactions that are required to disable thermodynamically infeasible energy production. CONCLUSION: Although not being fully comprehensive, our algorithm for systematic reaction direction assignment could define a significant number of irreversible reactions automatically with low computational effort. We envision that the presented algorithm is a valuable part of a computational framework that assists the automated reconstruction of genome-scale metabolic models. PMID- 17123435 TI - Comparative analysis of dinoflagellate chloroplast genomes reveals rRNA and tRNA genes. AB - BACKGROUND: Peridinin-containing dinoflagellates have a highly reduced chloroplast genome, which is unlike that found in other chloroplast containing organisms. Genome reduction appears to be the result of extensive transfer of genes to the nuclear genome. Unusually the genes believed to be remaining in the chloroplast genome are found on small DNA 'minicircles'. In this study we present a comparison of sets of minicircle sequences from three dinoflagellate species. RESULTS: PCR was used to amplify several minicircles from Amphidinium carterae so that a homologous set of gene-containing minicircles was available for Amphidinium carterae and Amphidinium operculatum, two apparently closely related peridinin-containing dinoflagellates. We compared the sequences of these minicircles to determine the content and characteristics of their chloroplast genomes. We also made comparisons with minicircles which had been obtained from Heterocapsa triquetra, another peridinin-containing dinoflagellate. These in silico comparisons have revealed several genetic features which were not apparent in single species analyses. The features include further protein coding genes, unusual rRNA genes, which we show are transcribed, and the first examples of tRNA genes from peridinin-containing dinoflagellate chloroplast genomes. CONCLUSION: Comparative analysis of minicircle sequences has allowed us to identify previously unrecognised features of dinoflagellate chloroplast genomes, including additional protein and RNA genes. The chloroplast rRNA gene sequences are radically different from those in other organisms, and in many ways resemble the rRNA genes found in some highly reduced mitochondrial genomes. The retention of certain tRNA genes in the dinoflagellate chloroplast genome has important implications for models of chloroplast-mitochondrion interaction. PMID- 17123436 TI - Feeding and fasting controls liver expression of a regulator of G protein signaling (Rgs16) in periportal hepatocytes. AB - BACKGROUND: Heterotrimeric G protein signaling in liver helps maintain carbohydrate and lipid homeostasis. G protein signaling is activated by binding of extracellular ligands to G protein coupled receptors and inhibited inside cells by regulators of G protein signaling (RGS) proteins. RGS proteins are GTPase activating proteins, and thereby regulate Gi and/or Gq class G proteins. RGS gene expression can be induced by the ligands they feedback regulate, and RGS gene expression can be used to mark tissues and cell-types when and where Gi/q signaling occurs. We characterized the expression of mouse RGS genes in liver during fasting and refeeding to identify novel signaling pathways controlling changes in liver metabolism. RESULTS: Rgs16 is the only RGS gene that is diurnally regulated in liver of ad libitum fed mice. Rgs16 transcription, mRNA and protein are up regulated during fasting and rapidly down regulated after refeeding. Rgs16 is expressed in periportal hepatocytes, the oxygen-rich zone of the liver where lipolysis and gluconeogenesis predominates. Restricting feeding to 4 hr of the light phase entrained Rgs16 expression in liver but did not affect circadian regulation of Rgs16 expression in the suprachiasmatic nuclei (SCN). CONCLUSION: Rgs16 is one of a subset of genes that is circadian regulated both in SCN and liver. Rgs16 mRNA expression in liver responds rapidly to changes in feeding schedule, coincident with key transcription factors controlling the circadian clock. Rgs16 expression can be used as a marker to identify and investigate novel G-protein mediated metabolic and circadian pathways, in specific zones within the liver. PMID- 17123437 TI - Comparing the efficacy of morphologic and DNA-based taxonomy in the freshwater gastropod genus Radix (Basommatophora, Pulmonata). AB - BACKGROUND: Reliable taxonomic identification at the species level is the basis for many biological disciplines. In order to distinguish species, it is necessary that taxonomic characters allow for the separation of individuals into recognisable, homogeneous groups that differ from other such groups in a consistent way. We compared here the suitability and efficacy of traditionally used shell morphology and DNA-based methods to distinguish among species of the freshwater snail genus Radix (Basommatophora, Pulmonata). RESULTS: Morphometric analysis showed that shell shape was unsuitable to define homogeneous, recognisable entities, because the variation was continuous. On the other hand, the Molecularly defined Operational Taxonomic Units (MOTU), inferred from mitochondrial COI sequence variation, proved to be congruent with biological species, inferred from geographic distribution patterns, congruence with nuclear markers and crossing experiments. Moreover, it could be shown that the phenotypically plastic shell variation is mostly determined by the environmental conditions experienced. CONCLUSION: Contrary to DNA-taxonomy, shell morphology was not suitable for delimiting and recognising species in Radix. As the situation encountered here seems to be widespread in invertebrates, we propose DNA-taxonomy as a reliable, comparable, and objective means for species identification in biological research. PMID- 17123438 TI - Ideal timing to transfer from an acute care hospital to an interdisciplinary inpatient rehabilitation program following a stroke: an exploratory study. AB - BACKGROUND: Timely accessibility to organized inpatient stroke rehabilitation services may become compromised since the demand for rehabilitation services following stroke is rapidly growing with no promise of additional resources. This often leads to prolonged lengths of stays in acute care facilities for individuals surviving a stroke. It is believed that this delay spent in acute care facilities may inhibit the crucial motor recovery process taking place shortly after a stroke. It is important to document the ideal timing to initiate intensive inpatient stroke rehabilitation after the neurological event. Therefore, the objective of this study was to examine the specific influence of short, moderate and long onset-admission intervals (OAI) on rehabilitation outcomes across homogeneous subgroups of patients who were admitted to a standardized interdisciplinary inpatient stroke rehabilitation program. METHODS: A total of 418 patients discharged from the inpatient neurological rehabilitation program at the Montreal Rehabilitation Hospital Network after a first stroke (79% of all cases reviewed) were included in this retrospective study. After conducting a matching procedure across these patients based on the degree of disability, gender, and age, a total of 40 homogeneous triads (n = 120) were formed according to the three OAI subgroups: short (less than 20 days), moderate (between 20 and 40 days) or long (over 40 days; maximum of 70 days) OAI subgroups. The rehabilitation outcomes (admission and discharge Functional Independence Measure scores (FIM), absolute and relative FIM gain scores, rehabilitation length of stay, efficiency scores) were evaluated to test for differences between the three OAI subgroups. RESULTS: Analysis revealed that the three OAI subgroups were comparable for all rehabilitation outcomes studied. No statistical difference was found for admission (P = 0.305-0.972) and discharge (P = 0.083-0.367) FIM scores, absolute (P = 0.533-0.647) and relative (P = 0.496 0.812) FIM gain scores, rehabilitation length of stay (P = 0.096), and efficiency scores (P = 0.103-0.674). CONCLUSION: OAI does not seem to affect significantly inpatient stroke rehabilitation outcomes of patients referred from acute care facilities where rehabilitation services are rapidly initiated after the onset of the stroke and offered throughout their stay. However, other studies considering factors such as the type and intensity of the rehabilitation are required to support those results. PMID- 17123439 TI - Changes in heart failure medications in patients hospitalised and discharged. AB - BACKGROUND: To date, evidence-based recommendations help doctors to manage patients with heart failure (HF). However, the implementation of these recommendations in primary care is still problematic as beneficial drugs are infrequently prescribed. The aim of the study was to determine whether admission to hospital increases usage of beneficial HF medication and if this usage is maintained directly after discharge. METHODS: The study was conducted from November 2002 until January 2004. In 77 patients hospitalised with heart failure (HF), the medication prescribed by the referring general practitioner (GP) and drug treatment directed by the hospital physicians was documented. Information regarding the post-discharge (14 d) therapy by the GP was evaluated via a telephone interview. Ejection fraction values, comorbidity and specifics regarding diagnostic or therapeutic intervention were collected by chart review. RESULTS: When compared to the referring GPs, hospital physicians prescribed more ACE-inhibitors (58.4% vs. 76.6%; p = 0.001) and beta-blockers of proven efficacy in HF (metoprolol, bisoprolol, carvedilol; 58.4% vs. 81.8%). Aldosterone antagonists were also administered more frequently in the hospital setting compared to general practice (14.3% vs. 37.7%). The New York Heart Association classification for heart failure did not influence whether aldosterone antagonists were administered either in primary or secondary care. Fourteen days after discharge, there was no significant discontinuity in discharge medication. CONCLUSION: Patients suffering from HF were more likely to receive beneficial medication in hospital than prior to admission. The treatment regime then remained stable two weeks after discharge. We suggest that findings on drug continuation in different cardiovascular patients might be considered validated for patients with HF. PMID- 17123440 TI - The glycolytic pathway of Trimastix pyriformis is an evolutionary mosaic. AB - BACKGROUND: Glycolysis and subsequent fermentation is the main energy source for many anaerobic organisms. The glycolytic pathway consists of ten enzymatic steps which appear to be universal amongst eukaryotes. However, it has been shown that the origins of these enzymes in specific eukaryote lineages can differ, and sometimes involve lateral gene transfer events. We have conducted an expressed sequence tag (EST) survey of the anaerobic flagellate Trimastix pyriformis to investigate the nature of the evolutionary origins of the glycolytic enzymes in this relatively unstudied organism. RESULTS: We have found genes in the Trimastix EST data that encode enzymes potentially catalyzing nine of the ten steps of the glycolytic conversion of glucose to pyruvate. Furthermore, we have found two different enzymes that in principle could catalyze the conversion of phosphoenol pyruvate (PEP) to pyruvate (or the reverse reaction) as part of the last step in glycolysis. Our phylogenetic analyses of all of these enzymes revealed at least four cases where the relationship of the Trimastix genes to homologs from other species is at odds with accepted organismal relationships. Although lateral gene transfer events likely account for these anomalies, with the data at hand we were not able to establish with confidence the bacterial donor lineage that gave rise to the respective Trimastix enzymes. CONCLUSION: A number of the glycolytic enzymes of Trimastix have been transferred laterally from bacteria instead of being inherited from the last common eukaryotic ancestor. Thus, despite widespread conservation of the glycolytic biochemical pathway across eukaryote diversity, in a number of protist lineages the enzymatic components of the pathway have been replaced by lateral gene transfer from disparate evolutionary sources. It remains unclear if these replacements result from selectively advantageous properties of the introduced enzymes or if they are neutral outcomes of a gene transfer 'ratchet' from food or endosymbiotic organisms or a combination of both processes. PMID- 17123441 TI - Combination phenylbutyrate/gemcitabine therapy effectively inhibits in vitro and in vivo growth of NSCLC by intrinsic apoptotic pathways. AB - BACKGROUND: Standard chemotherapy protocols in NSCLC are of limited clinical benefit. Histone deacetylase (HDAC) inhibitors represent a new strategy in human cancer therapy. In this study the combination of the HDAC inhibitor phenylbutyrate (PB) and the nucleoside analogue gemcitabine (GEM) was evaluated and the mechanisms underlying increased cell death were analyzed. METHODS: Dose escalation studies evaluating the cytotoxicity of PB (0.01-100 mM), GEM (0.01-100 microg/ml) and a combination of the two were performed on two NSCLC cell lines (BEN and KNS62). Apoptotic cell death was quantified. The involvement of caspase dependent cell death and MAP-kinase activation was analyzed. Additionally, mitochondrial damage was determined. In an orthotopic animal model the combined effect of PB and GEM on therapy was analyzed. RESULTS: Applied as a single drug both GEM and PB revealed limited potential to induce apoptosis in KNS62 and Ben cells. Combination therapy was 50-80% (p = 0.012) more effective than either agent alone. On the caspase level, combination therapy significantly increased cleavage of the pro-forms compared to single chemotherapy. The broad spectrum caspase-inhibitor zVAD was able to inhibit caspase cleavage completely, but reduced the frequency of apoptotic cells only by 30%. Combination therapy significantly increased changes in MTP and the release of cyto-c, AIF and Smac/Diabolo into the cytoplasm. Furthermore, the inhibitors of apoptosis c-IAP1 and c-IAP2 were downregulated and it was shown that in combination therapy JNK activation contributed significantly to induction of apoptosis. The size of the primary tumors growing orthotopically in SCID mice treated for 4 weeks with GEM and PB was significantly reduced (2.2-2.7 fold) compared to GEM therapy alone. The Ki-67 (KNS62: p = 0.015; Ben: p = 0.093) and topoisomerase IIalpha (KNS62: p = 0.008; Ben: p = 0.064) proliferation indices were clearly reduced in tumors treated by combination therapy, whereas the apoptotic index was comparably low in all groups. CONCLUSION: Therapy combining GEM and the HDAC inhibitor PB initiates a spectrum of apoptosis-inducing mitochondrial and further JNK-dependent events, thereby overcoming the therapeutic resistance of NSCLC tumor cells. In vivo, the combination therapy substantially reduced tumor cell proliferation, suggesting that the well tolerated PB is a useful supplemental therapeutic agent in NSCLC. PMID- 17123442 TI - Daily rhythm of salivary and serum urea concentration in sheep. AB - BACKGROUND: In domestic animals many biochemical and physiological processes exhibit daily rhythmicity. The aim of the present study was to investigate the rhythmic pattern of salivary and serum urea concentrations in sheep. METHODS: Six 3-year-old female sheep kept in the same environmental conditions were used. Sheep were sampled at 4 hour intervals for 48 consecutive hours starting at 08:00 of the first day and finishing at 04:00 of the second day. Blood samples were collected via intravenous cannulae inserted into the jugular vein; saliva samples were collected through a specific tube, the "Salivette". Salivary and serum urea concentrations were assayed by means of UV spectrophotometer. ANOVA was used to determine significant differences. The single Cosinor procedure was applied to the results showing significant differences over time. RESULTS: ANOVA showed a significant effect of time on salivary and serum urea concentrations. Serum and salivary urea peaked during the light phase. In the dark phase serum and salivary urea concentrations decreased, and the diurnal trough occurred at midnight. Cosinor analysis showed diurnal acrophases for salivary and serum urea concentrations. Daily mean levels were significantly higher in the serum than in the saliva. CONCLUSION: In sheep both salivary and serum urea concentrations showed daily fluctuations. Urea is synthesized in the liver and its production is strongly influenced by food intake. Future investigation should clarify whether daily urea rhythms in sheep are endogenous or are simply the result of the temporal administration of food. PMID- 17123443 TI - Bruxism secondary to brain injury treated with botulinum toxin-A: a case report. AB - We report a successful treatment of bruxism in a patient with anoxic brain injury using botulinum toxin-A (BTX-A). On examination the mouth opening was 0 mm, no feeding was possible through the mouth. Botulinum toxin was injected into the masseter and temporalis; great improvement in trismus and bruxism was noted after 3 weeks. One further treatment improved the mouth opening on the following week and the patient was discharged from our care to be reviewed when required. PMID- 17123444 TI - Genetic differentiation in the soil-feeding termite Cubitermes sp. affinis subarquatus: occurrence of cryptic species revealed by nuclear and mitochondrial markers. AB - BACKGROUND: Soil-feeding termites are particularly interesting models for studying the effects of fragmentation, a natural or anthropic phenomenon described as promoting genetic differentiation. However, studying the link between fragmentation and genetics requires a method for identifying species unambiguously, especially when morphological diagnostic characters are lacking. In humivorous termites, which contribute to the fertility of tropical soils, molecular taxonomy and phylogenetic relationships are rarely studied, though mitochondrial and nuclear molecular markers are widely used in studies of pest termites. Here, we attempt to clarify the taxonomy of soil-feeding colonies collected throughout the naturally fragmented Lope Reserve area (Gabon) and morphologically affiliated to Cubitermes sp. affinis subarquatus. The mitochondrial gene of cytochrome oxidase II (COII), the second nuclear rDNA internal transcribed spacer (ITS2) and five microsatellites were analyzed in 19 colonies. RESULTS: Bayesian Inference, Maximum Likelihood and Maximum Parsimony phylogenetic analyses, which were applied to the COII and ITS2 sequences, and Neighbor-Joining reconstructions, applied to the microsatellite data, reveal four major lineages in the Cubitermes sp. affinis subarquatus colonies. The concordant genealogical pattern of these unlinked markers strongly supports the existence of four cryptic species. Three are sympatric in the Reserve and are probably able to disperse within a mosaic of forests of variable ages and savannahs. One is limited to a very restricted gallery forest patch located in the North, outside the Reserve. CONCLUSION: Our survey highlights the value of combined mitochondrial and nuclear markers for exploring unknown groups such as soil feeding termites, and their relevance for resolving the taxonomy of organisms with ambiguous morphological diagnostic characters. PMID- 17123445 TI - Nutritional supplements and infection in the elderly: why do the findings conflict? AB - BACKGROUND: Most of the randomized placebo-controlled trials that have examined the clinical effects of multivitamin-mineral supplements on infection in the elderly have shown no significant effect. The exceptions are three such trials, all using a supplement with the same composition, and all claiming dramatic benefits: a frequently cited study published in 1992, which reported a 50% reduction in the number of days of infection (NDI), and two 2002 replication studies. Questions have been raised about the 1992 report; a second report in 2001 based on the same trial, but describing effects of the supplement on cognitive functions, has been retracted by Nutrition. The primary purpose of the present paper is to evaluate the claims about the effects of supplements on NDI in the two replication reports. METHODS: Examination of internal consistency (outcomes of statistical tests versus reported data); comparison of variability of NDI across individuals in these two reports with variability in other trials; estimation of the probability of achieving the reported close agreement with the original finding. RESULTS: The standard deviations of NDI and levels of statistical significance reported are profoundly inconsistent. The reported standard deviations of NDI are consistently below what other studies have found. The reported percent reductions in NDI agree too closely with the original study. CONCLUSION: The claims of reduced NDI in the two replication reports should be questioned, which also adds to concerns about the 1992 study. It follows that there is currently no trustworthy evidence from randomized placebo-controlled clinical trials that favors the use of vitamin-mineral supplements to reduce infection in the elderly. PMID- 17123446 TI - Family time, parental behaviour model and the initiation of smoking and alcohol use by ten-year-old children: an epidemiological study in Kaunas, Lithuania. AB - BACKGROUND: Family is considered to be the first and the most important child development and socialization bond. Nevertheless, parental behaviour model importance for the children, as well as family time for shared activity amount influence upon the child's health-related behaviour habit development has not been yet thoroughly examined. The aim of this paper is to indicate the advanced health-hazardous behaviour modelling possibilities in the families, as well as time spent for joint family activities, and to examine the importance of time spent for joint family activities for the smoking and alcohol use habit initiation among children. METHODS: This research was carried out in Kaunas, Lithuania, during the school year 2004-2005. The research population consisted of 369 fifth-grade schoolchildren (211 (57.2%) boys and 158 (42.8%) girls) and 565 parents: 323 (57.2%) mothers and 242 (48.2%) fathers. The response rate was 80.7% for children; 96.1% and 90.6% for mothers and fathers correspondingly. RESULTS: Eating a meal together was the most frequent joint family activity, whereas visiting friends or relatives together, going for a walk, or playing sports were the most infrequent joint family activities. More than two thirds (81.5%) of parents (248 (77.0%) mothers and 207 (85.9%) fathers (p < 0.05)) reported frequenting alcohol furnished parties at least once a month. About half of the surveyed fathers (50.6%) together with one fifth of the mothers (19.9%) (p < 0.001) were smokers. More frequently than girls, boys reported having tried smoking (6.6% and 23.0% respectively; p < 0.001) as well as alcohol (31.16% and 40.1% respectively; p < 0.05). Child alcohol use was associated both with paternal alcohol use, and with the time, spent in joint family activities. For instance, boys were more prone to try alcohol, if their fathers frequented alcohol furnished parties, whereas girls were more prone to try alcohol, if family members spent less time together. CONCLUSION: Joint family activity time deficit together with frequent parental examples of smoking and alcohol use underlie the development of alcohol and smoking addictions in children to some extent. The above-mentioned issues are suggested to be widely addressed in the comprehensive family health education programs. PMID- 17123447 TI - Evaluation of methods for extraction of the volitional EMG in dynamic hybrid muscle activation. AB - BACKGROUND: Hybrid muscle activation is a modality used for muscle force enhancement, in which muscle contraction is generated from two different excitation sources: volitional and external, by means of electrical stimulation (ES). Under hybrid activation, the overall EMG signal is the combination of the volitional and ES-induced components. In this study, we developed a computational scheme to extract the volitional EMG envelope from the overall dynamic EMG signal, to serve as an input signal for control purposes, and for evaluation of muscle forces. METHODS: A "synthetic" database was created from in-vivo experiments on the Tibialis Anterior of the right foot to emulate hybrid EMG signals, including the volitional and induced components. The database was used to evaluate the results obtained from six signal processing schemes, including seven different modules for filtration, rectification and ES component removal. The schemes differed from each other by their module combinations, as follows: blocking window only, comb filter only, blocking window and comb filter, blocking window and peak envelope, comb filter and peak envelope and, finally, blocking window, comb filter and peak envelope. RESULTS AND CONCLUSION: The results showed that the scheme including all the modules led to an excellent approximation of the volitional EMG envelope, as extracted from the hybrid signal, and underlined the importance of the artifact blocking window module in the process. The results of this work have direct implications on the development of hybrid muscle activation rehabilitation systems for the enhancement of weakened muscles. PMID- 17123448 TI - A technique to train new oculomotor behavior in patients with central macular scotomas during reading related tasks using scanning laser ophthalmoscopy: immediate functional benefits and gains retention. AB - BACKGROUND: Reading with a central scotoma involves the use of preferred retinal loci (PRLs) that enable both letter resolution and global viewing of word. Spontaneously developed PRLs however often privilege spatial resolution and, as a result, visual span is commonly limited by the position of the scotoma. In this study we designed and performed the pilot trial of a training procedure aimed at modifying oculomotor behavior in subjects with central field loss. We use an additional fixation point which, when combined with the initial PRL, allows the fulfillment of both letter resolution and global viewing of words. METHODS: The training procedure comprises ten training sessions conducted with the scanning laser ophthalmoscope (SLO). Subjects have to read single letters and isolated words varying in length, by combining the use of their initial PRL with the one of an examiner's selected trained retinal locus (TRL). We enrolled five subjects to test for the feasibility of the training technique. They showed stable maculopathy and persisting major reading difficulties despite previous orthoptic rehabilitation. We evaluated ETDRS visual acuity, threshold character size for single letters and isolated words, accuracy for paragraphed text reading and reading strategies before, immediately after SLO training, and three months later. RESULTS: Training the use of multiple PRLs in patients with central field loss is feasible and contributes to adapt oculomotor strategies during reading related tasks. Immediately after SLO training subjects used in combination with their initial PRL the examiner's selected TRL and other newly self-selected PRLs. Training gains were also reflected in ETDRS acuity, threshold character size for words of different lengths and in paragraphed text reading. Interestingly, subjects benefited variously from the training procedure and gains were retained differently as a function of word length. CONCLUSION: We designed a new procedure for training patients with central field loss using scanning laser ophthalmoscopy. Our initial results on the acquisition of newly self-selected PRLs and the development of new oculomotor behaviors suggest that the procedure aiming primarily at developing an examiner's selected TRL might have initiated a more global functional adaptation process. PMID- 17123449 TI - JUICE: a data management system that facilitates the analysis of large volumes of information in an EST project workflow. AB - BACKGROUND: Expressed sequence tag (EST) analyses provide a rapid and economical means to identify candidate genes that may be involved in a particular biological process. These ESTs are useful in many Functional Genomics studies. However, the large quantity and complexity of the data generated during an EST sequencing project can make the analysis of this information a daunting task. RESULTS: In an attempt to make this task friendlier, we have developed JUICE, an open source data management system (Apache + PHP + MySQL on Linux), which enables the user to easily upload, organize, visualize and search the different types of data generated in an EST project pipeline. In contrast to other systems, the JUICE data management system allows a branched pipeline to be established, modified and expanded, during the course of an EST project. The web interfaces and tools in JUICE enable the users to visualize the information in a graphical, user-friendly manner. The user may browse or search for sequences and/or sequence information within all the branches of the pipeline. The user can search using terms associated with the sequence name, annotation or other characteristics stored in JUICE and associated with sequences or sequence groups. Groups of sequences can be created by the user, stored in a clipboard and/or downloaded for further analyses. Different user profiles restrict the access of each user depending upon their role in the project. The user may have access exclusively to visualize sequence information, access to annotate sequences and sequence information, or administrative access. CONCLUSION: JUICE is an open source data management system that has been developed to aid users in organizing and analyzing the large amount of data generated in an EST Project workflow. JUICE has been used in one of the first functional genomics projects in Chile, entitled "Functional Genomics in nectarines: Platform to potentiate the competitiveness of Chile in fruit exportation". However, due to its ability to organize and visualize data from external pipelines, JUICE is a flexible data management system that should be useful for other EST/Genome projects. The JUICE data management system is released under the Open Source GNU Lesser General Public License (LGPL). JUICE may be downloaded from http://genoma.unab.cl/juice_system/ or http://www.genomavegetal.cl/juice_system/. PMID- 17123450 TI - A decade of change in breastfeeding in China's far north-west. AB - BACKGROUND: There have been considerable changes in breastfeeding practices in China over the past forty years. However China is a very large country, and breastfeeding rates in different parts of China vary considerably. The objective of this paper is to identify and compare breastfeeding types and rates between 1994-1996 and 2003-2004 in Shihezi, Xinjiang Uygur Autonomous Region, PR China. METHODS: In 1994-1996, a study of breastfeeding (n = 2197) was undertaken in Shihezi, Xinjiang, PR China. A decade later in 2003-2004, a longitudinal study (n = 545) of infant feeding practices was undertaken in the same area. RESULTS: The 'any breastfeeding' rates at 1, 4 and 6 months were 94%, 82% and 78% respectively in the early 1990s. A decade later, breastfeeding at 1 month was lower, but rates at 4 and 6 months remained the same. In 2004 the 'full breastfeeding' rate at one month was significantly higher (57%) than a decade earlier (38%), but after 3 months there was a rapid decline. This reflected a shift in the way complementary foods are introduced: the initial introduction was later, but by a higher proportion of mothers. CONCLUSION: The rate of breastfeeding at one month is significantly lower in 2003-2004 when compared to 1994-1996. The 'full breastfeeding' rates were initially higher, but after 3 months were then lower. The Chinese national breastfeeding targets were not reached in either period of the study. These studies show the need to further promote full or exclusive breastfeeding and further longitudinal studies are necessary to provide the detailed knowledge about risk factors required for health promotion programs. PMID- 17123451 TI - Duodenoportal fistula caused by peptic ulcer after extended right hepatectomy for hilar cholangiocarcinoma. AB - BACKGROUND: A fistula between the duodenum and the main portal vein near a peptic ulcer is extremely rare, and only two cases of duodenal ulcers have been reported in the past. CASE PRESENTATION: We report a 68-year-old man with a diagnosis of anemia who had a history of extended right hepatectomy for hilar cholangiocarcinoma 20 months previously. The first endoscopic examination revealed a giant peptic ulcer with active bleeding at the posterior wall of the duodenal bulbs, and hemostasis was performed. Endoscopic treatment and transarterial embolization were performed repeatedly because of uncontrollable bleeding from the duodenal ulcer. Nevertheless, he died of sudden massive hematemesis on the 20th hospital day. At autopsy, communication with the main portal vein and duodenal ulcer was observed. CONCLUSION: It should be borne in mind that the main portal vein is exposed at the front of the hepatoduodenal ligament in cases with previous extrahepatic bile duct resection. PMID- 17123452 TI - Pathway analysis of kidney cancer using proteomics and metabolic profiling. AB - BACKGROUND: Renal cell carcinoma (RCC) is the sixth leading cause of cancer death and is responsible for 11,000 deaths per year in the US. Approximately one-third of patients present with disease which is already metastatic and for which there is currently no adequate treatment, and no biofluid screening tests exist for RCC. In this study, we have undertaken a comprehensive proteomic analysis and subsequently a pathway and network approach to identify biological processes involved in clear cell RCC (ccRCC). We have used these data to investigate urinary markers of RCC which could be applied to high-risk patients, or to those being followed for recurrence, for early diagnosis and treatment, thereby substantially reducing mortality of this disease. RESULTS: Using 2-dimensional electrophoresis and mass spectrometric analysis, we identified 31 proteins which were differentially expressed with a high degree of significance in ccRCC as compared to adjacent non-malignant tissue, and we confirmed some of these by immunoblotting, immunohistochemistry, and comparison to published transcriptomic data. When evaluated by several pathway and biological process analysis programs, these proteins are demonstrated to be involved with a high degree of confidence (p values < 2.0 E-05) in glycolysis, propanoate metabolism, pyruvate metabolism, urea cycle and arginine/proline metabolism, as well as in the non-metabolic p53 and FAS pathways. In a pilot study using random urine samples from both ccRCC and control patients, we performed metabolic profiling and found that only sorbitol, a component of an alternative glycolysis pathway, is significantly elevated at 5.4-fold in RCC patients as compared to controls. CONCLUSION: Extensive pathway and network analysis allowed for the discovery of highly significant pathways from a set of clear cell RCC samples. Knowledge of activation of these processes will lead to novel assays identifying their proteomic and/or metabolomic signatures in biofluids of patient at high risk for this disease; we provide pilot data for such a urinary bioassay. Furthermore, we demonstrate how the knowledge of networks, processes, and pathways altered in kidney cancer may be used to influence the choice of optimal therapy. PMID- 17123453 TI - Malignant melanomas of the nasolacrimal duct. AB - Two patients with a history of epistaxis who were both found to have a nasolacrimal duct melanoma are presented. A literature review revealed that no previous cases of primary nasolacrimal duct melanoma have been reported. Current therapeutic modalities are discussed. PMID- 17123454 TI - Advancement in post-meningitic lateral semicircular canal labyrinthitis ossificans. AB - OBJECTIVE: To assess whether lateral semicircular canal (LSCC) ossification is more advanced than that in the cochlear basal turn, in order to judge the value of the former as a predictor. METHODS: Retrospective review of 33 paediatric patients from our cochlear implant programme, with profound sensorineural hearing loss after bacterial meningitis. Magnetic resonance imaging (MRI), computed tomography (CT) scans and operative findings were reviewed. RESULTS: On CT, LSCC ossification scores were more advanced than those for the cochlear basal turn in 69.9 per cent of implanted ears. Forty-five per cent (15/33) of children had ossification at surgery. In predicting this, the sensitivity of CT LSCC ossification was 90 per cent and that of MRI LSCC ossification was 83.3 per cent. CONCLUSIONS: The more advanced ossification found in the LSCC, compared with that in the cochlear basal turn, adds to previous findings of LSCC pathology predicting cochlear ossification. Surprisingly, CT of the LSCC appears to be no less valuable than MRI in pre-operative cochlear implant assessment of post meningitic children. PMID- 17123455 TI - Temporal order in memory and interval timing: an interference analysis. AB - The effect of varying load in memory tasks performed during a time interval production was examined. In a first experiment, increasing load in memory search for temporal order affected concurrent time production more strongly than varying load in a spatial memory task of equivalent difficulty. This result suggests that timing uses some specific resources also required in processing temporal order in memory, resources that would not be used in the spatial memory task. A second experiment showed that the interference between time production and memory search involving temporal order was stronger when, during the timing task, a decision was made on the temporal position of a memory item, than when information on temporal order was retained throughout the interval to be produced. These results underscore the importance of considering the specific resources and processes involved when the interference between timing and concurrent non-temporal tasks is analyzed. PMID- 17123456 TI - A microbial diagnostic microarray technique for the sensitive detection and identification of pathogenic bacteria in a background of nonpathogens. AB - A major challenge in microbial diagnostics is the parallel detection and identification of low-bundance pathogens within a complex microbial community. In addition, a high specificity providing robust, reliable identification at least at the species level is required. A microbial diagnostic microarray approach, using single nucleotide extension labeling with gyrB as the marker gene, was developed. We present a novel concept applying competitive oligonucleotide probes to improve the specificity of the assay. Our approach enabled the sensitive and specific detection of a broad range of pathogenic bacteria. The approach was tested with a set of 35 oligonucleotide probes targeting Escherichia coli, Shigella spp., Salmonella spp., Aeromonas hydrophila, Vibrio cholerae, Mycobacterium avium, Mycobacterium tuberculosis, Helicobacter pylori, Proteus mirabilis, Yersinia enterocolitica, and Campylobacter jejuni. The introduction of competitive oligonucleotides in the labeling reaction successfully suppressed cross-reaction by closely related sequences, significantly improving the performance of the assay. Environmental applicability was tested with environmental and veterinary samples harboring complex microbial communities. Detection sensitivity in the range of 0.1% has been demonstrated, far below the 5% detection limit of traditional microbial diagnostic microarrays. PMID- 17123457 TI - Electrospun poly(vinylidene fluoride)/poly(aminophenylboronic acid) composite nanofibrous membrane as a novel glucose sensor. AB - Electrospinning was used to prepare the nanofibrous membrane (NFM) of the composite comprising poly(vinylidene fluoride) and poly(aminophenylboronic acid) (PVdF/PAPBA-NFM). The PVdF/PAPBA-NFM displayed an excellent linear response to the detection of glucose for the concentration range of 1 to 15mM with a response time of less than 6s. Further experiments on amperometric sensing of glucose were performed in the presence of interferents such as uric acid, ascorbic acid, acetaminophen, fructose, mannose, etc. using PVdF/PAPBA-NFM. The interferents did not give significant overlapping current signal during the determination of glucose. Also, PVdF/PAPBA-NFM possesses better reproducibility toward glucose detection and storage stability. PMID- 17123458 TI - Regulation of cation channels in cardiac and smooth muscle cells by intracellular magnesium. AB - Magnesium regulates various ion channels in many tissues, including those of the cardiovascular system. General mechanisms by which intracellular Mg(2+) (Mg(i)(2+)) regulates channels are presented. These involve either a direct interaction with the channel, or an indirect modification of channel function via other proteins, such as enzymes or G proteins, or via membrane surface charges and phospholipids. To provide an insight into the role of Mg(i)(2+) in the cardiovascular system, effects of Mg(i)(2+) on major channels in cardiac and smooth muscle cells and the underlying mechanisms are then reviewed. Although Mg(i)(2+) concentrations are known to be stable, conditions under which they may change exist, such as following stimulation of beta-adrenergic receptors and of insulin receptors, or during pathophysiological conditions such as ischemia, heart failure or hypertension. Modifications of cardiovascular electrical or mechanical function, possibly resulting in arrhythmias or hypertension, may result from such changes of Mg(i)(2+) and their effects on cation channels. PMID- 17123459 TI - Acetylcholine synthesis, muscarinic receptor subtypes, neuropeptides and secretion of ferret salivary glands with special reference to the zygomatic gland. AB - Studies on salivary secretion are usually focused on parotid and submandibular glands. However, the film of mucin, that protects the oral structures and is responsible for the feeling of oral comfort, is produced by the submucosal glands. The submucosal zygomatic and molar glands are particularly large in carnivores such as the ferret. Comparisons between the mucous sublingual, zygomatic and molar glands, serous parotid and sero-mucous submandibular glands showed the acetylcholine synthesis, in terms of concentration, to be three to four times higher in the mucous glands than in the parotid and submandibular glands. Bromoacetylcholine inhibited 95-99% of the synthesis of acetylcholine in the incubates of the five types of glands, showing the acetylcholine synthesis to depend on the activity of choline acetyltransferase. The high acetylcholine synthesis in the zygomatic gland was of nervous origin, since cutting the buccal nerve, aiming at parasympathetic denervation, and allowing time for nerve degeneration, reduced the acetylcholine synthesising capacity of the gland by 95%. A similar reduction (96%) in the parotid gland followed upon the avulsion of the parasympathetic auriculo-temporal nerve. Zygomatic saliva was very viscous. The salivary flow rate in response to electrical stimulation (20 Hz) of the buccal nerve (zygomatic gland), expressed per gland weight, was one-third of that to stimulation of the auriculo-temporal nerve (parotid gland) or the chorda lingual nerve (submandibular gland). As previously shown for the parotid and submandibular gland, a certain fraction (25%) of the parasympathetic secretory response of the zygomatic gland depended on non-adrenergic, non-cholinergic transmission mechanisms, probably involving substance P and vasoactive intestinal peptide and possibly calcitonin gene-related peptide. Particularly, high concentrations of vasoactive intestinal peptide were found in the sublingual and molar glands, and of substance P in the submandibular, zygomatic and molar glands; notably, the concentration of calcitonin gene-related peptide of the sublingual gland was not detectable. All five muscarinic receptor subtypes were detected in the five glands. The receptor protein profile, as judged by immunoblotting and semi-quantitative estimations, was about the same in the glands: high level of M3, low level of M2 and levels roughly in the same range of M1, M4 and M5. Compared to the parotid and submandibular glands, the M5 receptor level was particularly low in the mucin-secreting glands. The present study points out both similarities and dissimilarities between the five types of glands investigated. The zygomatic gland, in particular, appears to be a suitable model for future studies aiming at causing relief of dry mouth by local pharmacological treatment. PMID- 17123460 TI - Bongkrekic acid and atractyloside inhibits chloride channels from mitochondrial membranes of rat heart. AB - The aim of this work was to characterize the effect of bongkrekic acid (BKA), atractyloside (ATR) and carboxyatractyloside (CAT) on single channel properties of chloride channels from mitochondria. Mitochondrial membranes isolated from a rat heart muscle were incorporated into a bilayer lipid membrane (BLM) and single chloride channel currents were measured in 250/50 mM KCl cis/trans solutions. BKA (1-100 microM), ATR and CAT (5-100 microM) inhibited the chloride channels in dose-dependent manner. The inhibitory effect of the BKA, ATR and CAT was pronounced from the trans side of a BLM and it increased with time and at negative voltages (trans-cis). These compounds did not influence the single channel amplitude, but decreased open dwell time of channels. The inhibitory effect of BKA, ATR and CAT on the mitochondrial chloride channel may help to explain some of their cellular and/or subcellular effects. PMID- 17123461 TI - On the advantages of using green light to study fluorescence yield changes in leaves. AB - In photosynthetic chains, the kinetics of fluorescence yield depends on the photochemical rates at the level of both Photosystem I and II and thus on the absorption cross section of the photosynthetic units as well as on the coupling between light harvesting complexes and photosynthetic traps. A new set-up is described which, at variance with the commonly used set-ups, uses of a weakly absorbed light source (light-emitting diodes with maximum output at 520 nm) to excite the photosynthetic electron chain and probe the resulting fluorescence yield changes and their time course. This approach optimizes the homogeneity of the exciting light throughout the leaf and we show that this homogeneity narrows the distribution of the photochemical rates. Although the exciting light is weakly absorbed, the possibility to tune the intensity of the light emitting diodes allows one to reach photochemical rates ranging from 10(4) s(-1) to 0.25 s(-1) rendering experimentally accessible different functional regimes. The variations of the fluorescence yield induced by the photosynthetic activity are qualitatively and quantitatively discussed. When illuminating dark-adapted leaves by a weak light, the kinetics of fluorescence changes displays a pronounced plateau which precedes the fluorescence increase reflecting the full reduction of the plastoquinone pool. We ascribe this plateau to the time delay needed to reduce the photosystem I electron acceptors. PMID- 17123462 TI - CO-dependent H2 evolution by Rhodospirillum rubrum: role of CODH:CooF complex. AB - Upon exposure to CO during anaerobic growth, the purple phototrophic bacterium Rhodospirillum rubrum expresses a CO-oxidizing H(2) evolving enzymatic system. The CO-oxidizing enzyme, carbon monoxide dehydrogenase (CODH), has been purified and extensively characterized. However the electron transfer pathway from CODH to the CO-induced hydrogenase that evolves H(2) is not well understood. CooF is an Fe-S protein that is the proposed mediator of electron transfer between CODH and the CO-induced hydrogenase. Here we present the spectroscopic and biochemical properties of the CODH:CooF complex. The characteristic EPR signals observed for CODH are largely insensitive to CooF complexation. Metal analysis and EPR spectroscopy show that CooF contains 2 Fe(4)S(4) clusters. The observation of 2 Fe(4)S(4) clusters for CooF contradicts the prediction of 4 Fe(4)S(4) clusters based on analysis of the amino acid sequence of CooF and structural studies of CooF homologs. Comparison of in vivo and in vitro CO-dependent H(2) evolution indicates that approximately 90% of the activity is lost upon cell lysis. We propose that the loss of two labile Fe-S clusters from CooF during cell lysis may be responsible for the low in vitro CO-dependent H(2) evolution activity. During the course of these studies, a new assay for CODH:CooF was developed using membranes from an R. rubrum mutant that did not express CODH:CooF, but expressed high levels of the CO-induced hydrogenase. The assay revealed that the CO-induced hydrogenase requires the presence of CODH:CooF for optimal H(2) evolution activity. PMID- 17123463 TI - Function of two beta-carotenes near the D1 and D2 proteins in photosystem II dimers. AB - The antenna proteins in photosystem II (PSII) not only promote energy transfer to the photosynthetic reaction center (RC) but provide also an efficient cation sink to re-reduce chlorophyll a if the electron transfer (ET) from the Mn-cluster is inhibited. Using the newest PSII dimer crystal structure (3.0 A resolution), in which 11 beta-carotene molecules (Car) and 14 lipids are visible in the PSII monomer, we calculated the redox potentials (Em) of one-electron oxidation for all Car (Em(Car)) by solving the Poisson-Boltzmann equation. In each PSII monomer, the D1 protein harbors a previously unlocated Car (CarD1) in van der Waals contact with the chlorin ring of ChlZ(D1). Each CarD1 in the PSII dimer complex is located in the interface between the D1 and CP47 subunits, together with another four Car of the other PSII monomer and several lipid molecules. The proximity of Car bridging between CarD1 and plastoquinone/Q(A) may imply a direct charge recombination of Car+Q(A)-. The calculated Em(CarD1) and Em(ChlZ(D1)) are, respectively, 83 and 126 mV higher than Em(CarD2) and Em(ChlZ(D2)), which could explain why CarD2+ and ChlZ(D2)+ are observed rather than the corresponding CarD1+ and ChlZ(D1)+. PMID- 17123464 TI - Deciphering the mechanisms of intestinal imino (and amino) acid transport: the redemption of SLC36A1. AB - The absorption of zwitterionic imino and amino acids, and related drugs, is an essential function of the small intestinal epithelium. This review focuses on the physiological roles of transporters recently identified at the molecular level, in particular SLC36A1, by identifying how they relate to the classical epithelial imino and amino acid transporters characterised in mammalian small intestine in the 1960s-1990s. SLC36A1 transports a number of D- and L-imino and amino acids, beta- and gamma-amino acids and orally-active neuromodulatory and antibacterial agents. SLC36A1 (or PAT1) functions as a proton-coupled imino and amino acid symporter in cooperation with the Na+/H+ exchanger NHE3 (SLC9A3) to produce the imino acid carrier identified in rat small intestine in the 1960s but subsequently ignored because of confusion with the IMINO transporter. However, it is the sodium/imino and amino acid cotransporter SLC6A20 which corresponds to the betaine carrier (identified in hamster, 1960s) and IMINO transporter (identified in rabbit and guinea pig, 1980s). This review summarises evidence for expression of SLC36A1 and SLC6A20 in human small intestine, highlights the differences in functional characteristics of the imino acid carrier and IMINO transporter, and explains the confusion surrounding these two distinct transport systems. PMID- 17123465 TI - Blood-injection-injury fears: harm- vs. disgust-relevant selective outcome associations. AB - There is increasing evidence that blood-injection-injury (BII) phobia is qualitatively different from the other specific phobias in the sense that phobic distress takes the form of disgust rather than (threat-induced) fear. Following this, we tested the relative importance of harm and disgust-related associative biases in BII-fear. High (n=25) and low (n=27) fearful individuals saw a series of fear-relevant (blood-related) and fear-irrelevant (rabbit and flower) slides which were randomly paired with either a harm-related outcome, a disgust-related outcome, or nothing. Preexperimentally, participants expected blood-related slides to be followed by both disgust- and harm-relevant outcomes. These selective preexperimental outcome expectancies were readily corrected during the experiment. Neither low nor high fearful participants showed a postexperimental covariation bias. The absence of differential effects between high and low fearful participants does not support the idea that disgust- or harm-relevant associative biases play a role in the maintenance of BII-fears. The results corroborate the previous finding of Pury and Mineka [1997. Covariation bias for blood-injury stimuli and aversive outcomes. Behaviour Research and Therapy, 35, 35-47] that people are generally liable to selectively associate BII-stimuli with aversive outcomes. PMID- 17123466 TI - High penetrance of sequencing errors and interpretative shortcomings in mtDNA sequence analysis of LHON patients. AB - For identifying mutation(s) that are potentially pathogenic it is essential to determine the entire mitochondrial DNA (mtDNA) sequences from patients suffering from a particular mitochondrial disease, such as Leber hereditary optic neuropathy (LHON). However, such sequencing efforts can, in the worst case, be riddled with errors by imposing phantom mutations or misreporting variant nucleotides, and moreover, by inadvertently regarding some mutations as novel and pathogenic, which are actually known to define minor haplogroups. Under such circumstances it remains unclear whether the disease-associated mutations would have been determined adequately. Here, we re-analyse four problematic LHON studies and propose guidelines by which some of the pitfalls could be avoided. PMID- 17123467 TI - Novel substrate specificity of glutathione synthesis enzymes from Streptococcus agalactiae and Clostridium acetobutylicum. AB - Glutathione (GSH) is synthesized by gamma-glutamylcysteine synthetase (gamma-GCS) and glutathione synthetase (GS) in living organisms. Recently, bifunctional fusion protein, termed gamma-GCS-GS catalyzing both gamma-GCS and GS reactions from gram-positive firmicutes Streptococcus agalactiae, has been reported. We revealed that in the gamma-GCS activity, S. agalactiae gamma-GCS-GS had different substrate specificities from those of Escherichia coli gamma-GCS. Furthermore, S. agalactiae gamma-GCS-GS synthesized several kinds of gamma-glutamyltripeptide, gamma-Glu-X(aa)-Gly, from free three amino acids. In Clostridium acetobutylicum, the genes encoding gamma-GCS and putative GS were found to be immediately adjacent by BLAST search, and had amino acid sequence homology with S. agalactiae gamma-GCS-GS, respectively. We confirmed that the proteins expressed from each gene showed gamma-GCS and GS activity, respectively. C. acetobutylicum GS had broad substrate specificities and synthesized several kinds of gamma glutamyltripeptide, gamma-Glu-Cys-X(aa). Whereas the substrate specificities of gamma-GCS domain protein and GS domain protein of S. agalactiae gamma-GCS-GS were the same as those of S. agalactiae gamma-GCS-GS. PMID- 17123468 TI - Dicoumarol impairs mitochondrial electron transport and pyrimidine biosynthesis in human myeloid leukemia HL-60 cells. AB - Dicoumarol, a competitive inhibitor of NAD(P)H:quinone oxidoreductase 1 (NQO1), increases intracellular superoxide and affects cell growth of tumor cells. This work was set to establish a mechanistic link between dicoumarol, superoxide and cell cycle alterations in HL-60 cells. Using ES936, a mechanism-based irreversible inhibitor of NQO1, we demonstrate that NQO1 inhibition is not a major factor involved in superoxide boost. Mitochondrial Complexes II, III and IV were directly inhibited by dicoumarol. Succinate, which inhibits superoxide generation by reversed electron flow in Complex II, significantly decreased superoxide boost in dicoumarol-treated cells and in isolated mitochondria incubated with dicoumarol and decylubiquinol. Superoxide generation in cells was strongly potentiated by blocking the quinone site of Complex II with thenoyltrifluoroacetone, supporting the involvement of cytochrome b560 to drive electrons for increasing superoxide. Simultaneous inhibition of the mitochondrial chain upstream ubiquinone and displacement of succinate from the Complex II active site is proposed as a major mechanism to explain how dicoumarol increases superoxide in HL-60 cells. Dicoumarol-treated cells accumulated in S phase due to the impairment of pyrimidine biosynthesis at dihydroorotate dehydrogenase step because blockade was overcome by addition of exogenous uridine or orotate, but not by dihydroorotate. We demonstrate for the first time that dicoumarol inhibits mitochondrial electron transport, induces superoxide release by reversed electron flow in Complex II, and inhibits pyrimidines biosynthesis. These actions must be taken into account when considering dicoumarol effects on cells. PMID- 17123469 TI - The metabolomics of (+/-)-arecoline 1-oxide in the mouse and its formation by human flavin-containing monooxygenases. AB - The alkaloid arecoline is a main constituent of areca nuts that are chewed by approximately 600 million persons worldwide. A principal metabolite of arecoline is arecoline 1-oxide whose metabolism has been poorly studied. To redress this, synthetic (+/-)-arecoline 1-oxide was administered to mice (20mg/kg p.o.) and a metabolomic study performed on 0-12h urine using ultra-performance liquid chromatography-coupled time-of-flight mass spectrometry (UPLC-TOFMS) with multivariate data analysis. A total of 16 mass/retention time pairs yielded 13 metabolites of (+/-)-arecoline 1-oxide, most of them novel. Identity of metabolites was confirmed by tandem mass spectrometry. The principal pathways of metabolism of (+/-)-arecoline 1-oxide were mercapturic acid formation, with catabolism to mercaptan and methylmercaptan metabolites, apparent CC double-bond reduction, carboxylic acid reduction to the aldehyde (a novel pathway in mammals), N-oxide reduction, and de-esterification. Relative percentages of metabolites were determined directly from the metabolomic data. Approximately, 50% of the urinary metabolites corresponded to unchanged (+/-)-arecoline 1-oxide, 25% to other N-oxide metabolites, while approximately, 30% corresponded to mercapturic acids or their metabolites. Many metabolites, principally mercapturic acids and their derivatives, were excreted as diastereomers that could be resolved by UPLC-TOFMS. Arecoline was converted to arecoline 1-oxide in vitro by human flavin-containing monooxygenases FMO1 (K(M): 13.6+/-4.9muM; V(MAX): 0.114+/ 0.01nmolmin(-1)microg(-1) protein) and FMO3 (K(M): 44.5+/-8.0microM; V(MAX): 0.014+/-0.001nmolmin(-1)microg(-1) protein), but not by FMO5 or any of 11 human cytochromes P450. This report underscores the power of metabolomics in drug metabolite mining. PMID- 17123470 TI - Combined dexamethasone/corticotropin releasing hormone test predicts treatment response in major depression - a potential biomarker? AB - BACKGROUND: Exaggerated corticotropin (ACTH) and cortisol response to the combined dexamethasone (DEX)/corticotropin releasing hormone (CRH) test, indicating impaired regulation of the hypothalamus-pituitary-adrenocortical (HPA) system, is frequently observed in depression. In the present study, we examined whether change in HPA system function during the first weeks of hospitalization predicts response to antidepressant treatment in major depression and thus constitutes a potential biomarker. METHODS: We conducted the DEX/CRH test in 50 inpatients suffering from severe major depression, once after study inclusion and a second time 2 to 3 weeks later while under continuous antidepressant treatment. RESULTS: We found increased ACTH and cortisol responses to the first DEX/CRH test compared with healthy control subjects. In the second DEX/CRH test 2 to 3 weeks later, 36 of the 50 patients showed an attenuated cortisol response, while 14 patients did not display improvement or exhibited even aggravation of the altered HPA system function. Improved HPA system regulation in the second DEX/CRH test was associated with beneficial treatment response after 5 weeks and a higher remission rate at the end of hospitalization. CONCLUSIONS: The results suggest that change in HPA system regulation assessed with repeated DEX/CRH tests is a potential biomarker that may predict clinical outcome at follow-up. There is consensus that the drug development process could be improved, once reliable biomarkers become available that help to allow a judgement regarding the efficacy of a novel drug candidate. The combined DEX/CRH test seems to be a promising candidate for such a biomarker. PMID- 17123471 TI - Reduced amygdala volumes in first-episode bipolar disorder and correlation with cerebral white matter. AB - BACKGROUND: Previous magnetic resonance imaging (MRI) findings on amygdala volume abnormalities in bipolar disorder have been inconsistent, which may partly reflect clinical heterogeneity. It is unclear whether amygdala abnormalities are present early in the course of illness and/or are the consequence of disease progression. METHODS: Twenty patients with first-episode bipolar disorder and 23 matched healthy comparison subjects were included. Magnetic resonance images were used to measure amygdala volumes, as well as whole brain measures of gray and white matter volume. RESULTS: First-episode bipolar patients had significant reductions in amygdala volume relative to healthy subjects in an analysis of covariance that accounted for the effects of age, sex, and whole brain volume. First-episode patients also showed a trend reduction in cerebral white matter volume, and there was a significant correlation between cerebral white matter volume and total amygdala volume in patients but not control subjects. CONCLUSIONS: These findings indicate that amygdala volume deficits are present early in the course of bipolar disorder and may occur within a neuroanatomical context of reduced cerebral white matter. Additional research should examine whether the nature of regional white matter deficits, particularly in frontal temporal tracts, may help parse the pathophysiology of amygdala volume abnormalities in bipolar disorder. PMID- 17123472 TI - Decreased serum levels of epidermal growth factor in adult subjects with high functioning autism. AB - BACKGROUND: The neurobiological basis for autism remains poorly understood. Given the role of growth factors in brain development, we hypothesized that epidermal growth factor (EGF) may play a role in the pathophysiology of autism. In this study, we examined whether serum levels of EGF are altered in adult subjects with high-functioning autism. METHODS: We measured serum levels of EGF in the 17 male subjects with high-functioning autism and 18 age-matched healthy male subjects. RESULTS: The serum levels of EGF in the subjects with high-functioning autism (72.4 +/- 102.8 pg/mL [mean +/- SD]) were significantly lower (Mann-Whitney U = 22.0, p < .001) than those of normal control subjects (322.3 +/- 122.0 pg/mL [mean +/- SD]). However, there were no correlations between serum EGF levels and clinical variables in the subjects with autism. CONCLUSIONS: This study suggests that decreased levels of EGF might be implicated in the pathophysiology of high functioning autism. PMID- 17123473 TI - Analysis of association between the serotonin transporter and antidepressant response in a large clinical sample. AB - BACKGROUND: SLC6A4 encodes the serotonin transporter, the protein primarily responsible for the termination of serotonin neurotransmission. Because many antidepressants inhibit the transporter, it has been the focus of intense pharmacogenetic analysis. We sought to replicate our previous findings that SLC6A4 is associated with response to a selective serotonin reuptake inhibitor (SSRI) in a large case-control study. METHODS: Genotypes at the SLC6A4 locus were obtained for 1,914 subjects in the Sequenced Treatment Alternatives to Relieve Depression (STAR*D) study and then tested for association to treatment response of the SSRI citalopram. RESULTS: Nine tagging single nucleotide polymorphisms and two variants previously associated with antidepressant response, including a promoter repeat polymorphism, were genotyped. Single marker and haplotypic analyses failed to detect association with antidepressant response in the largest clinical sample studied to date. CONCLUSIONS: The lack of association between response to an SSRI and variation at the SLC6A4 locus in this large sample, carefully characterized for response to citalopram, strongly suggests that SSRI response in major depression is not determined by DNA variation at this locus. These findings do not replicate findings of a number of studies with considerably smaller sample sizes. Other genetic determinants of SSRI response in depression should be sought. PMID- 17123474 TI - Genetic and environmental predictors of early alcohol use. AB - BACKGROUND: The goal of the current investigation was to examine genetic and environmental predictors of early alcohol use, a potent predictor of later alcohol dependence. METHODS: This study represents an add-on project to an investigation examining the efficacy of an intervention for maltreated children entering out-of-home care. Predictors of early alcohol use include the following: maltreatment, family loading for alcohol or substance-use disorders, and serotonin transporter genotype (5-HTTLPR; locus SLC6A4). Participants included 127 subjects: 76 maltreated children and 51 demographically matched community controls. RESULTS: At follow-up, 29% of the maltreated children reported alcohol use, a rate more than seven times the rate observed in controls. Maltreated children also drank alcohol, on average, more than 2 years earlier than controls (11.2 vs. 13.5 years). Early alcohol use was predicted by maltreatment, 5-HTTLPR, and a gene by environment interaction, with increased risk for early alcohol use associated with the s-allele. Psychopathology at baseline, severity of maltreatment, and poor mother-child relations also predicted early alcohol use. CONCLUSIONS: Maltreated children are at high risk for psychiatric, alcohol, and substance abuse problems. Examination of genetic and environmental risk and protective factors can help identify those who are most vulnerable and help guide prevention and intervention efforts. PMID- 17123475 TI - Prenatal, perinatal, and postnatal risk factors in obsessive-compulsive disorder. AB - BACKGROUND: The etiology of obsessive-compulsive disorder (OCD) remains unknown, although it is thought to involve an interaction of genetic and environmental factors. This study aimed to identify prenatal, perinatal, and postnatal risk factors in OCD. METHODS: We compared retrospectively 68 OCD patients to 70 control subjects based on responses given on a standardized questionnaire. The questionnaire was designed to evaluate environmental factors, with a special focus on gestation, labor, birth, and early infancy aspects. RESULTS: The group of OCD patients had risk factors with greater frequency than the control group. Notable among the significant findings (p < or = 0.001) were edema of the hands, feet, or face and excessive weight gain during gestation; hyperemesis gravidarum; prolonged labor; preterm birth; and jaundice. When socioeconomic class was used as a covariable in the logistic regression analysis, prolonged labor and edema during pregnancy remained statistically significant. CONCLUSIONS: Some early risk factors may be associated with the expression of OCD later in life such as edema during pregnancy and prolonged labor. If our findings are confirmed in future studies, greater attention should be given to such factors in predisposed individuals, especially in prenatal care and delivery. PMID- 17123476 TI - P50 auditory evoked potential suppression in bipolar disorder patients with psychotic features and their unaffected relatives. AB - BACKGROUND: Diminished suppression of the P50 response, a consistent finding in schizophrenia, has also been reported in patients with psychotic bipolar disorder. It is a promising endophenotype for schizophrenia, but its relationship to genetic liability in bipolar disorder is unknown. We therefore assessed whether diminished P50 suppression is associated with familial risk for psychotic bipolar disorder. METHODS: The P50 response was collected in a conditioning (C)- testing (T) paradigm from 42 outpatients with bipolar 1 disorder who had experienced psychotic symptoms and 44 of their unaffected first-degree relatives, all from families multiply affected with bipolar disorder or another non-organic psychotic disorder; 48 healthy control subjects were also studied. The T/C ratio was compared between the groups, with linear regression analyses and robust variance estimators for clustered data. RESULTS: Both patients (estimated mean difference in T/C ratio to control subjects, 32, 95% confidence interval [CI] 15 48, p=.001) and unaffected relatives (20, 95% CI 7-32, p=.002) demonstrated higher T/C ratio, thus indicating diminished P50 suppression compared with control subjects. CONCLUSIONS: To our knowledge, this is the first report of diminished P50 gating in unaffected relatives of psychotic bipolar disorder patients from multiply affected families. Our results suggest that impaired P50 gating is a putative endophenotype for psychotic bipolar disorder and thus might reflect the impact of susceptibility genes across psychosis. PMID- 17123477 TI - Anatomical evidence of impaired feedforward auditory processing in schizophrenia. AB - BACKGROUND: Somal volumes of pyramidal cells are reduced within feedforward but not feedback circuits in areas 41 and 42 of the auditory cortex of subjects with schizophrenia. Because neuronal somal volume depends on both the number of axonal terminations onto and furnished by the neuron, we hypothesized that axon terminal densities are reduced in feedforward but not feedback auditory pathways in subjects with schizophrenia. METHODS: We used stereologic methods to quantify the density of a marker of axon terminals, synaptophysin-immunoreactive (SY-IR) puncta, in areas 41 and 42 of 15 subjects with schizophrenia and matched normal comparison subjects. The effect of long-term haloperidol exposure on density of SY-IR puncta was similarly evaluated in nonhuman primates. RESULTS: Synaptophysin immunoreactive puncta density was 13.6% lower in deep layer 3 of area 41 in the schizophrenia subjects but was not changed in layer 1 of area 41 or in deep layer 3 of area 42. Density of SY-IR puncta did not differ between haloperidol-exposed and control monkeys. CONCLUSIONS: Reduction of SY-IR puncta density is selective for feedforward circuits within primary auditory cortex of subjects with schizophrenia. This deficit may contribute to impairments in auditory sensory processing in this disorder. PMID- 17123479 TI - Levels-of-processing effects in first-degree relatives of individuals with schizophrenia. AB - BACKGROUND: First-degree relatives of individuals with schizophrenia show cognitive impairments that are similar to but less severe than their ill relatives. We have shown that memory impairments can be improved and prefrontal cortical (PFC) activity increased in individuals with schizophrenia by providing beneficial encoding strategies. The current study used a similar paradigm to determine whether siblings of individuals with schizophrenia (SIBs) also show increases in brain activity when presented with beneficial encoding strategies. METHODS: Twenty-one SIBs and 38 siblings of healthy comparison subjects underwent functional magnetic resonance imaging scans while engaged in deep (abstract/concrete judgments) and shallow (orthographic judgments) encoding. Subjects were then given a recognition memory test. RESULTS: The groups did not differ on encoding or recognition accuracy, and the SIBs benefited from deep encoding to a similar degree as control subjects. The SIBs showed deep encoding related activity in a number of PFC regions typically activated during semantic processing. However, SIBs showed more activity than control subjects in three subregions of PFC (left BA 44 & BA 47 bilaterally). CONCLUSIONS: Siblings of individuals with schizophrenia benefit from supportive verbal encoding conditions. Like individuals with schizophrenia, SIBs also show increased task related activity in a larger number of PFC subregions than control subjects during deep verbal encoding. PMID- 17123478 TI - White-matter integrity predicts stroop performance in patients with geriatric depression. AB - BACKGROUND: This study tested the hypothesis that microstructural white matter abnormalities in frontostriatal-limbic tracts are associated with poor response inhibition on the Stroop task in depressed elders. METHOD: Fifty-one elders with major depression participated in a 12-week escitalopram trial. Diffusion tensor imaging was used to determine fractional anisotropy (FA) in white matter regions. Executive function (response inhibition) was assessed with the Stroop task. Voxelwise correlational analysis was used to examine the relationship between Stroop performance and fractional anisotropy. RESULTS: Significant associations between FA and Stroop color word interference were evident in multiple frontostriatal-limbic regions, including white matter lateral to the anterior and posterior cingulate cortex and white matter in prefrontal, insular, and parahippocampal regions. CONCLUSIONS: These findings suggest that microstructural white matter abnormalities of frontostriatal-limbic networks are associated with executive dysfunction of late-life depression. This observation provides the rationale for examination of specific frontostriatal-limbic pathways in the pathophysiology of geriatric depression. PMID- 17123480 TI - Hippocampal volume and mood disorders after traumatic brain injury. AB - BACKGROUND: Recent evidence from clinical studies and animal models of traumatic brain injury (TBI) suggest that neuronal and glial loss might progress after the initial insult in selectively vulnerable regions of the brain such as the hippocampus. There is also evidence that hippocampal dysfunction plays a role in the pathogenesis of mood disorders. We examined the relationship between hippocampal damage and mood disorders after TBI and the effect of hippocampal atrophy on the outcome of TBI patients. METHODS: The study group consisted of 37 patients with closed head injury who were evaluated at baseline and at 3, 6, and 12 months after trauma. Psychiatric diagnosis was made with a structured clinical interview and DSM-IV criteria. Quantitative magnetic resonance imaging scans were obtained at 3-months follow-up. RESULTS: Patients with moderate to severe head injury had significantly lower hippocampal volumes than patients with mild TBI. Patients who developed mood disorders had significantly lower hippocampal volumes than patients without mood disturbance. Furthermore, there was a significant interaction between mood disorders diagnosis and severity of TBI, by which patients with moderate to severe TBI who developed mood disorders had significantly smaller hippocampal volumes than patients with equivalent severe TBI who did not develop mood disturbance. Finally, reduced hippocampal volumes were associated with poor vocational outcome at 1-year follow-up. CONCLUSIONS: Our findings are consistent with a "double-hit" mechanism by which neural and glial elements already affected by trauma are further compromised by the functional changes associated with mood disorders (e.g., the neurotoxic effects of increased levels of cortisol or excitotoxic damage resulting from overactivation of glutaminergic pathways). Finally, patients with greater hippocampal damage were less likely to return to a productive life 1 year after trauma. PMID- 17123481 TI - Effect of schizotypy on cognitive performance and its tuning by COMT val158 met genotype variations in a large population of young men. AB - BACKGROUND: Mirroring schizophrenia, specific dimensions of schizotypy are related to cognitive dysfunction. The relation of schizotypy and state psychopathology to cognitive performance and its link to catechol-O methyltransferase (COMT) val(158) met genotype variations was studied in a large sample of young men. METHODS: State psychopathology and schizotypy were assessed with self-rated questionnaires. Cognitive performance was assessed with tests of reasoning ability, sustained attention, and verbal and spatial working memory. Subjects were genotyped for the val(158) met polymorphism of the gene for COMT (low enzymatic activity met/met, intermediate met/val, and high val/val). RESULTS: The val/val group had higher scores in measures of state psychopathology as well as negative and disorganized schizotypy dimensions, whereas there was no effect of COMT genotype on cognitive performance measures. Structural equation modeling showed that cognitive performance accuracy but not speed decreased with increasing negative schizotypy, increased with increasing paranoid schizotypy, and was not affected by state psychopathology. Increasing val loading resulted in a dose-dependent increase in the factor loading for the relation between negative schizotypy and cognitive performance accuracy. CONCLUSIONS: Different schizotypal phenotypes had opposing relations to cognitive performance in the population. COMT genotype modulated the relation between the negative schizotypal phenotype and cognitive performance. PMID- 17123482 TI - A protocol for the differentiation of human embryonic stem cells into dopaminergic neurons using only chemically defined human additives: Studies in vitro and in vivo. AB - Our ability to use human embryonic stem (hES) cells in cell replacement therapy for Parkinson's disease depends on the discovery of ways to simply and reliably differentiate a dopaminergic (DA) phenotype in these cells. Although several protocols exist for the differentiation of DA traits in hES, they involve the prolonged use of complex media with undefined components, cell conditioned media and/or co-culture with various cells, usually of animal origin. In this study, several well-characterized (H9, BG01) and several new uncharacterized (HUES7, HUES8) hES cell lines were studied for their capacity to differentiate into DA neurons in culture using a novel rapid protocol which uses only chemically defined human-derived media additives and substrata. Within 3 weeks, cells from all 4 cell lines progressed from the undifferentiated state to beta-tubulin III positive cells expressing DA markers in vitro. Moreover, transplantation of these cells into the striata of 6-hydroxydopamine-treated rats at the neuronal progenitor stage resulted in the appearance of differentiated DA traits in vivo 2 3 weeks later. PMID- 17123483 TI - Neuronal protein 22 colocalises with both the microtubule and microfilament cytoskeleton in neurite-like processes. AB - The expression of human neuronal protein 22 (hNP22) is up-regulated in the superior frontal cortex of chronic alcoholics. hNP22 shares significant homology with a number of proteins implicated in bundling of actin filaments. In addition, it contains domains similar to those found in microtubule-associated proteins. We investigated the ability of hNP22 to induce cytoskeletal changes by overexpression in Chinese hamster ovary cells. Overexpression of hNP22 resulted in process formation in these cells that increased upon treatment with cytochalasin D, an actin depolymerising agent. Transfection of mutant hNP22 containing either a deletion of the putative actin-binding domain or deletion of a consensus protein kinase C (PKC) phosphorylation site (Ser-180) failed to induce process formation. In contrast, a mutation to mimic persistent PKC phosphorylation resulted in a cellular morphology similar to that seen in wild type hNP22 transfections. This observation suggests that hNP22 requires phosphorylation at Ser-180 by PKC to induce cytoskeletal rearrangements. hNP22 was also observed to colocalise with actin and tubulin in processes of transfected cells. An hNP22-specific antibody specifically immunoprecipitated a complex including tubulin from human brain indicating that hNP22 binds directly to microtubules. Taken together, this data suggests that NP22 is part of a signaling complex that associates with cytoskeletal elements to regulate neuronal morphology. PMID- 17123484 TI - Changes in neuropeptide Y protein expression following photothrombotic brain infarction and epileptogenesis. AB - This study characterized morphological changes in the cortex and hippocampus of Sprague-Dawley rats following photothrombotic infarction and epileptogenesis with emphasis on the distribution of neuropeptide Y (NPY) expression. Animals were lesioned in the left sensorimotor cortex and compared with age-matched naive and sham-operated controls by immunohistochemical techniques at 1, 3, 7, and 180 days post-lesioning (DPL). NPY immunostaining was assessed by light microscopy and quantified by the optical fractionator technique using unbiased stereological methods. At 1, 3, and 7 DPL, the number of NPY-positive somata in the lesioned cortex was increased significantly compared to controls and the contralateral cortex. At 180 DPL, lesioned epileptic animals with frequent seizure activity demonstrated significant increases of NPY expression in the cortex, CA1, CA3, hilar interneurons, and granule cells of the dentate gyrus. In addition to NPY immunostaining, neuronal degeneration, cell death/cell loss, and astroglial response were assessed with cell-specific markers. Nissl and NeuN staining showed reproducible infarctions at each investigated time point. FJB-positive somata were most abundant in the infarct core at 1 DPL, decreased markedly at 3 DPL, and virtually absent by 7 DPL. Activated astroglia were detected in the cortex and hippocampus following lesioning and the development of seizure activity. In summary, NPY protein expression and morphological changes following cortical photothrombosis were time-, region-, and pathologic state-dependent. Alterations in NPY expression may reflect reactive or compensatory responses of the rat brain to acute infarction and to the development and expression of epileptic seizures. PMID- 17123485 TI - Effect of prenatal exposure to an anti-inflammatory drug on neuron number in cornu ammonis and dentate gyrus of the rat hippocampus: a stereological study. AB - Prenatal exposed to an anti-inflammatory drug is a major problem for the developing central nervous system. It is not well known the effect of prenatal exposed to a non-steroidal anti-inflammatory drug on the hippocampus. Total neuron number in one side of the cornu ammonis (CA) and gyrus dentatus (GD) of the hippocampal formation in control and drug-treated (diclofenac sodium, DS) groups of male rats was estimated using the optical fractionator technique. Each main group has also two subgroups that are 4 weeks old (4W-old) and 20 weeks old (20W-old). In CA, no significant difference between 4W-old DS-treated and their control was found, but a significant difference was observed between 20W-old DS treated and their controls. A decreasing of neuron number was 12% for 20W-old DS treated group. In GD, a decreasing of the granule cell number in 4W-old of DS treated group was seen but an increasing of granule cell number was found in the 20W-old drug-treated rats in comparison to its control group, 7% and 9%, respectively. Although an increasing of neuron number in CA at the control group was seen with age, from 4th week to 20th week (10%), age-dependent substantial granule cell decline (17%) was observed in GD. No age effect on the total cell numbers of CA and GD of the drug-treated groups was seen in comparison to 4W-old week and 20W-old. A pronounced neuron loss observed in the drug-treated group may be attributed to the neurotoxicity of diclofenac sodium (DS) on the developing hippocampal formation. Age-dependent neuron increase in the CA of 20W-old and neuron decline in GD of 20W-old control groups may be a result of a dual effect of saline injection during the fetal life, since these animals were exposed to a stress of 15-day-period of saline injection, prenatal stress. The reason of no age effect on CA and GD cell number in the drug-treated groups may be attributed to the depletion of the progenitor cells due to neurotoxicity of DS in the fetal life of these animals. PMID- 17123486 TI - Understanding words in context: the role of Broca's area in word comprehension. AB - What role does meaning selection play in word comprehension, and what neural systems support this selection process? Most words have multiple meanings and are therefore ambiguous. This is true of both homonymous words (words that have multiple unrelated meanings) and polysemous words (words that have multiple related meanings). The extant evidence indicates that meaning selection is an integral part of homonym comprehension. However, it is not known whether meaning selection extends to polysemous words, or what neural systems support meaning selection during comprehension. Prior neuroimaging and neuropsychological evidence suggest that the left inferior frontal gyrus (LIFG) may play a role in resolving competition during language processing. We therefore sought to test the hypotheses that meaning selection is part of polysemous word comprehension, and that the LIFG resolves meaning competition during word comprehension. We tested healthy participants on a version of the triplet lexical decision task, with polysemous and homonymous stimuli. Results suggest that the meanings of polysemous words, like the meanings of homonyms, are selected based on context. However, homonymous and polysemous words differed in how meaning frequency affected meaning selection. We then administered the triplet lexical decision task to patients with LIFG damage to examine whether this region plays a role in context-dependent meaning selection. Results support the hypothesis that the LIFG serves as a top-down biasing mechanism that facilitates rapid meaning selection during word comprehension. We conclude that context-dependent meaning selection is an integral part of word comprehension for both homonyms and polysemous words, and that the LIFG facilitates this selection process. PMID- 17123487 TI - Aquaporin expression in the cerebral cortex is increased at early stages of Alzheimer disease. AB - Abnormalities in the cerebral microvasculature are common in Alzheimer disease (AD). Expression levels of the water channels aquaporin 1 and aquaporin 4 (AQP1, AQP4) were examined in AD cases by gel electrophoresis and Western blotting, and densitometric values normalized with beta-actin were compared with corresponding values in age-matched controls processed in parallel. In addition, samples of cases with Pick disease (PiD) were examined for comparative purposes. A significant increase in the expression levels of AQP1 was observed in AD stage II (following Braak and Braak classification). Individual variations were seen in advanced stages which resulted in non-significant differences between AD stages V VI and age-matched controls. No differences in AQP1 levels were observed between familial AD cases (FAD, all of them at advanced stages) and corresponding age matched controls. Immunohistochemistry showed increased AQP1 in astrocytes at early stages of AD. Double-labelling immunofluorescence and confocal microscopy disclosed AQP1 immunoreactivity at the cell surface of astrocytes which were recognized with anti-glial fibrillary acidic protein antibodies. No differences in the levels of AQP4 were observed in AD, FAD and PiD when compared with corresponding controls. These results indicate abnormal expression of AQP1 in astrocytes in AD, and they add support to the idea that abnormal regulation of mechanisms involved in the control of water fluxes occurs at early stages in AD. PMID- 17123488 TI - Wide therapeutic time window for fasudil neuroprotection against ischemia-induced delayed neuronal death in gerbils. AB - The neuroprotective potential and therapeutic time window for fasudil, a Rho kinase inhibitor (RKI), were evaluated for delayed neuronal death in gerbils. A preliminary screening was done on fasudil, ozagrel, and edaravone using a single administration in a delayed neuronal death study. Intraperitoneal (i.p.) administration of edaravone, a free radical scavenger (3, 10 mg/kg) immediately after re-circulation did not reduce neuronal degeneration. We previously reported that ozagrel, a thromboxane A(2) synthetase inhibitor (30 mg/kg) also did not reduce neuronal degeneration, while fasudil (3, 30 mg/kg) significantly protected against the ischemia-induced neuronal loss. To clarify the therapeutic time window of fasudil, which showed a positive effect in a preliminary screening, animals received their first i.p. administration of fasudil (10 mg/kg) 24 or 48 h after ischemia. Administration of fasudil twice daily was continued until day 6. Fasudil significantly protected against the ischemia-induced delayed neuronal death when the treatment was started 24 h after ischemia. In gerbils, hydroxyfasudil, an active metabolite of fasudil, was found following an i.p. administration of fasudil (10 mg/kg), and the value of the area under the plasma level curve of hydroxyfasudil was 7 times higher than that of fasudil. Hydroxyfasudil may contribute to the potency of fasudil. The present findings indicate that the RKI fasudil reduces ischemic neuronal damage with a wide therapeutic time window in gerbil, and may be useful in the treatment of acute ischemic stroke in humans. PMID- 17123489 TI - Formation of covalent beta-linked carbohydrate-enzyme intermediates during the reactions catalyzed by alpha-amylases. AB - Porcine pancreatic and Bacillus amyloliquefaciens alpha-amylases were examined for the formation of covalent carbohydrate intermediates during reaction. The enzymes were precipitated and denatured by adding 10 volumes of acetone. When these denatured enzymes were mixed with methyl alpha-6-[(3)H] maltooligosaccharide glycosides and chromatographed on BioGel P-2, no carbohydrate was found in the protein void volume peak. When the enzymes were added to the methyl alpha-6-[(3)H]-maltooligosaccharide glycosides and allowed to react for 15s at 1 degrees C and then precipitated and denatured with 10 volumes of acetone, (3)H-labeled carbohydrates were found in the BioGel P-2 protein void volume peak, indicating the formation of enzyme-carbohydrate covalent intermediates. (1)H NMR analysis of the denatured enzyme from the reaction with methyl alpha-maltooligosaccharide glycosides confirmed that carbohydrate was attached to the denatured enzyme. (1)H NMR saturation-transfer analysis further showed that the carbohydrate was attached to the denatured enzyme by a beta configuration. This configuration is what would be expected for an enzyme that catalyzes the hydrolysis of alpha-(1-->4) glycosidic linkages by a two-step, S(N)2 double-displacement reaction to give retention of the alpha-configuration of the substrates at the reducing-end of the products. PMID- 17123490 TI - The structure of the core-O-chain linkage region of the lipopolysaccharide from Bordetella hinzii. AB - Linkage region between core and the O-chain of the lipopolysaccharide from Bordetella hinzii has been analyzed by NMR and MS analysis of the products, obtained by anhydrous HF treatment or consecutive ammonia and AcOH treatment of the LPS. The following structure of this region was deduced from the experimental results: [structure: see text] This structure is identical to the structure of the respective region of Bordetella parapertussis LPS. Polysaccharide part (PS) consists of not more than 15 2,3-diacetamido-2,3-dideoxyhexuronamides, methylated at the only hydroxyl group of the non-reducing terminal monosaccharide. PMID- 17123491 TI - The synthesis of cyclic imidates from amides of glucuronic acid and investigation of glycosidation reactions. AB - The synthesis of novel cyclic glycosyl imidates and an investigation of their potential as donors in glycosidation reactions is described. The results show that 1,2-cis glycosides obtained from the reactions of glycosyl acetates or cyclic imidates, each derived from amides of glucuronic acid, result from the anomerisation of initially formed 1,2-trans glycosides. PMID- 17123492 TI - Inhibitory effects of different antioxidants on hyaluronan depolymerization. AB - Hyaluronan (HA) was depolymerized by hydroxyl radicals generated from hydrogen peroxide and cupric ions. Inhibition of HA degradation by four well-known antioxidants was investigated, as HA can scavenge reactive oxygen species (ROS). Change in hyaluronan molecular weight was observed by size-exclusion chromatography. Inhibition of HA degradation was estimated from the retention times observed. It was found that HA degradation was inhibited in a clearly concentration-dependent manner by mannitol, thiourea and vinpocetine. Propofol also inhibited the depolymerization, but its concentration-dependent effect was not so clear. The antioxidant concentrations at which HA degradation was decreased by 50% were 42 microM for thiourea; 1.35 microM for vinpocetine; and 0.39 microM for propofol. A concentration of 26.51 mM of mannitol was needed to attain the same inhibitory effect. Although many factors are involved in a therapeutic response, the results obtained in this study support the idea that HA may be protected from ROS attack by the concomitant use of well-known antioxidants. PMID- 17123493 TI - Vitamin C prevents zidovudine-induced NAD(P)H oxidase activation and hypertension in the rat. AB - OBJECTIVE: Cardiovascular risk is increased among HIV-infected patients receiving antiretroviral therapy due to the development of hypertension and metabolic abnormalities. In this study, we investigated the effects of long-term treatment with zidovudine (AZT) and vitamin C, alone and in combination, on blood pressure and on the chain of events linking oxidative stress to cardiac damage in the rat. METHODS: Six adult Wistar Kyoto rats received AZT (1 mg/ml) in the drinking water for 8 months, six vitamin C (10 g/kg of food) and AZT, six vitamin C alone, and six served as controls. RESULTS: AZT increased systolic blood pressure, expression of gp91(phox) and p47(phox) subunits of NAD(P)H oxidase, and protein kinase C (PKC) delta activation and reduced antioxidant power of plasma and cardiac homogenates. AZT also caused morphological alterations in cardiac myocyte mitochondria, indicative of functional damage. All of these effects were prevented by vitamin C. CONCLUSION: Chronic AZT administration increases blood pressure and promotes cardiovascular damage through a NAD(P)H oxidase-dependent mechanism that involves PKC delta. Vitamin C antagonizes these adverse effects of AZT in the cardiovascular system. PMID- 17123494 TI - Human leukocytes express ephrinB2 which activates microvascular endothelial cells. AB - EphrinB2-EphB4 interaction modulates the migration/adhesion of various cell types, including endothelial cells (EC) and peripheral blood leukocytes (PBLs). We hypothesize that the Ephrin/Eph signaling mechanism plays a role in mediating EC/leukocyte interactions during inflammation. PBLs were isolated from human blood, stimulated with inflammatory mediators, and total RNA or protein assayed for EphrinB2 expression. PBLs demonstrated differential expression profiles of EphrinB2 mRNA or protein, depending on cell subtype and stimulus. Human iris tissue and iris EC (HIEC) were examined for the expression of EphB4 mRNA and protein. Some blood vessels were EphB4(+), while stimulation of purified HIEC did not alter their expression of EphB4. HIEC treated with sEphrinB2/Fc from 0 to 60min did exhibit changes in their phospho-Erk1/2 levels. These observations indicate that stimulated lymphocytes express EphrinB2, which has the potential to activate EC. This suggests a novel mechanism by which EC and lymphocytes communicate to regulate cell activation/migration during inflammation. PMID- 17123495 TI - 1,3-Butadiene and leukemia among synthetic rubber industry workers: exposure response relationships. AB - Previous research updated the mortality experience of North American synthetic rubber industry workers during the period 1944-1998, determined if leukemia and other cancers were associated with several employment factors and carried out Poisson regression analysis to examine exposure-response associations between estimated exposure to 1,3-butadiene (BD) or other chemicals and cancer. The present study used Cox regression procedures to examine further the exposure response relationship between several unlagged and lagged, continuous, time dependent BD exposure indices (BD parts per million (ppm)-years, the total number of exposures to BD concentrations >100 ppm ("peaks") and average intensity of BD) and leukemia, lymphoid neoplasms and myeloid neoplasms. All three BD exposure indices were associated positively with leukemia. Using continuous, untransformed BD ppm-years the regression coefficient (beta) from an analysis that controlled only for age was 2.9 x 10(-4) (p<0.01); the regression coefficient adjusted for all covariates (age, year of birth, race, plant, years since hire and dimethyldithiocarbamate) was similar in magnitude (beta=3.0 x 10(-4), p=0.04). Lagging exposure had minimal impact on the results for leukemia for any of the three BD exposure indices. In models that controlled only for age, lymphoid neoplasms were associated with BD ppm-years and myeloid neoplasms, with BD peaks, but neither trend was statistically significant after adjusting for multiple covariates. The present results support the presence of a causal relationship between high cumulative exposure and high intensity of exposure to BD and leukemia. PMID- 17123496 TI - Dietary caloric restriction improves the redox status at the onset of diabetes in hepatocytes of streptozotocin-induced diabetic rats. AB - Enhanced production of free radicals and oxidative stress induced by hyperglycemia play a central role in the pathogenesis of diabetes and its complications. This study assessed the attenuation by dietary caloric restriction on the oxidative and lipid peroxidative effects of diabetes in the liver through reduction in body and organ weights and concomitant metabolic changes. Three month-old male Wistar rats were subjected to ad libitum feeding and 30% caloric restriction for 9 weeks before induction of diabetes by intraperitoneal injection of 35 mg/kg body weight streptozotocin. The animals were sacrificed 2 weeks after streptozotocin treatment depicting the onset of diabetes. Caloric restriction significantly reduced the organ weights (p<0.01), malondialdehyde (p<0.01) and catalase activity (p<0.01), but significantly increased glutathione reductase activity (p<0.01), and GSH/GSSG ratios (p<0.05). Caloric restriction also non significantly reduced reactive oxygen species, superoxide dismutase and oxidized glutathione but increased glutathione peroxidase activity and reduced glutathione levels in the diabetic rats. Our data indicate a decrease in lipid peroxidation, improvement in the antioxidant defense systems and restoration of the redox status in the liver by caloric restriction. Therefore, this could provide a non invasive antioxidant therapy early in diabetes to prevent the development of the complications associated with the disease. PMID- 17123497 TI - 8-quinolyl phosphate as a substrate for the fluorimetric determination of alkaline phosphatase. AB - BACKGROUND: Alkaline phosphatase (ALP) is an important target for clinical analysis. 8-Quinolyl phosphate (QP) was developed as a new substrate for the fluorimetric determination of ALP activity. METHODS: QP is a strong fluorescent substance and the product of the enzyme reaction is 8-hydroxyquinoline (HQ), which has no fluorescence. Under the optimal conditions for the determination of ALP, the decreased fluorescence intensity via the enzyme reaction is proportional to ALP activity. The fluorescence intensity was measured at lambdaex/lambdaem=318/495 nm before and after the enzyme reaction. RESULTS: QP reacted with ALP in the buffer solution of pH=9.5 and incubated for 20 min at 37.0 degrees C were selected as the optimal conditions for the determination of ALP. The linear range and detection limit for the determination of ALP are 1.0 16.0 and 0.229 U/l, respectively. With this method, ALP could be applied to assess ALP in human serum and the results were evaluated by comparison with a standard colorimetric assay using p-nitrophenyl phosphate as ALP substrate. CONCLUSIONS: This method is simple, practical and can be used as an alternative to assess ALP in clinical analysis. PMID- 17123498 TI - Neutralizing antibodies to interferon beta in multiple sclerosis: analytical evaluation for validation of a cytopathic effect assay. AB - BACKGROUND: Recent guidelines have recommended the use of validated assays for the measurement of neutralizing antibodies (NABs) to interferon beta (IFNbeta) in patients with multiple sclerosis (MS). In an attempt of validation, we studied the analytical performance of a bioassay based on antiviral cytopathic effect (CPE) using WISH cells and the vesicular stomatitis virus (WISH/VSV CPE). METHODS: NAB titres measured with the WISH/VSV CPE assay in 63 sera from IFNbeta treated MS patients were compared to those obtained with the reference CPE method using A549 cells and the encephalomyocarditis virus. Binding antibodies (BABs) were measured using a capture ELISA as a screening test for NABs. RESULTS: No false-negative BAB was obtained in our patients. The between-run coefficients of variation (CVs) determined with log10 titres of the NIH anti-IFNbeta (G038-501 572) yielded good results (0.10). The sex difference in VMH volume was primarily accounted for by the ventrolateral subdivision (VMHvl), which in both females and TFM males was significantly smaller than in wild-type males (Ps<0.005). There was no significant sex difference in the volume of the other three subdivisions of the VMH. Neuronal somata were larger in males than females in VMHvl, central VMH (VMHc) and the dorsomedial VMH (VMHdm), with TFM males having feminine neuronal somata in the VMHdm and VMHc. These data suggest that AR plays a role during sexual differentiation of the VMH, imparting its greatest effect in the VMHvl. ARs may regulate aromatase expression or activity to affect estrogen receptor activation, or may act independently of estrogen receptors to influence VMH morphology. PMID- 17123533 TI - Sensitivity and specificity of the Clock Drawing Test, incorporating Rouleau scoring system, as a screening instrument for questionable and mild dementia: scale development. AB - OBJECTIVE: The objective of this study was to validate the Rouleau scoring system for the Clock Drawing Test (CDT) in northern Taiwan, a non-English speaking area, to increase its generalization by detecting subjects with questionable dementia (QD) from normal controls (NC) and subjects with mild dementia (AD). The system consists of two parts: a 10-point scoring and error types. A cross-sectional design was used. One hundred and sixteen subjects were recruited from a memory disorder clinic (n=40 NC; n=34 QD; n=42 AD). Reliability and validity were estimated, the predictive accuracy was calculated using the receiver operating characteristic (ROC) curve analysis, and the error types were counted. RESULTS: The results indicated that most of the error types committed were conceptual deficiencies and graphic difficulties; more errors were found in the command condition (CDT-command) than in the copy condition (CDT-copy); 15% of the subjects drew smaller clock circles in CDT-command, which did not relate to any other errors. No small clock was found in CDT-copy because a pre-dawn circle was provided. The inter-rater reliability of the Rouleau scoring system was .87 and .83 for the CDT-command and -copy, respectively, while the discriminate accuracy of the scoring system was relatively low in detecting QD vs. NC, QD vs. AD, but somewhat better in distinguishing AD vs. NC groups for the areas under the ROC curves was equal to .72 for the CDT-command, and .73 for the CDT-copy. Visuospatial construction and executive functioning explained the largest score variance of the CDT in both conditions, while depressive symptoms were not significantly associated with the CDT performance. CONCLUSION: The CDT using the Rouleau scoring system has been validated in mild AD subjects in Taiwan; however, to screen for QD sufferers, alternative scoring systems emphasizing hands or combining CDT with different neuropsychological tests such as the Clinical Abilities Screening Instrument are recommended. PMID- 17123534 TI - Mapping of the complement C9 binding domain in paramyosin of the blood fluke Schistosoma mansoni. AB - Schistosomes are believed to evade complement-mediated damage by expression of complement inhibitory proteins. Our previous results [Deng, J., Gold, D., LoVerde, P.T., Fishelson, Z., 2003. Inhibition of the complement membrane attack complex by Schistosoma mansoni paramyosin. Infect. Immun. 71, 6402-6410.] have demonstrated that paramyosin (Pmy) of the blood fluke S. mansoni binds to the human complement proteins C8 and C9, inhibits complement activation at the terminal stage and protects the parasite from complement-mediated damage. In order to locate the Pmy binding site to C8 and C9, various fragments of Pmy cDNA were PCR-cloned into a pET28a bacterial expression vector. Recombinant His-tagged Pmy fragments were expressed in BL21 Escherichia coli and purified over a nickel nitrilotriacetic acid column. Binding assays by Western blotting with monoclonal anti-His antibody demonstrated that PmyCC (Pmy amino acids (744)Asp-(866)Met) was the only Pmy fragment that bound to human C8 and C9. Functional analyses demonstrated that PmyCC inhibited hemolysis of rabbit erythrocytes and of antibody-sensitized sheep erythrocytes by human complement. Importantly, PmyCC inhibited in vitro killing of trypsin-sensitized schistosomula of S. mansoni by human complement. In the presence of PmyCC, Zn(2+)-induced C9 polymerization was inhibited. Most of the immunodominant B-cell antigenic epitopes of Pmy are present in the PmyCC region, as antibodies collected from mice immunized with recombinant Pmy bound primarily to PmyCC. Taken together, this study has mapped the complement regulatory domain in Pmy, capable of binding to C8 and C9 and preventing polyC9 formation, to its C-terminal region. PMID- 17123535 TI - Race (black-white) and gender divergences in the relationship of childhood cardiovascular risk factors to carotid artery intima-media thickness in adulthood: the Bogalusa Heart Study. AB - It has been established that childhood cardiovascular (CV) risk factors are predictive of adulthood vascular changes as measured by carotid intima-media thickness (IMT). However, whether this relationship is race- and gender-specific is not known. This aspect was examined in a black-white cohort of 868 adults (29% blacks, 42% males) aged 25-44 years who were examined at least twice in childhood for traditional CV risk factors with an average follow-up period of 26.4 years. The average value of the two earliest childhood measurements was used as the childhood value, standardized to age, race, and gender-specific z-score. Carotid IMT was measured by B-mode ultrasonography. The mean of the maximum carotid IMT readings of three right and three left far walls for common, bulb and internal segments was used. In univariate analysis, significant correlates of adulthood carotid IMT (standardized to age-, race- and gender-specific z-score) were, in the order of decreasing magnitude, triglyceride and LDL cholesterol in white males; systolic blood pressure, LDL cholesterol, and body mass index (BMI) in white females; systolic blood pressure in black males; BMI and systolic blood pressure in black females. In multivariate regression analysis, significant predictors of carotid IMT were triglycerides and LDL cholesterol in white males; systolic blood pressure and LDL cholesterol in white females; systolic blood pressure in black males; and BMI and LDL cholesterol in black females. In conclusion, the predictability of childhood CV risk factors for increased carotid IMT in adulthood varies by race and gender. The prevention implications of these findings need further investigation. PMID- 17123536 TI - Low plasma adiponectin exacerbates the risk of premature coronary artery disease in familial hypercholesterolemia. AB - Familial hypercholesterolemia (FH) is characterized by increased risk for premature coronary artery disease (CAD). This risk is exacerbated in the presence of abdominal obesity and insulin resistance. Low adiponectin is part of the clustering of metabolic abnormalities associated with abdominal obesity and insulin resistance. The present study, therefore, aims to examine the relationship between plasma adiponectin and age at CAD diagnosis in FH patients. Plasma adiponectin was measured by ELISA in 568 non-diabetic FH individuals of French-Canadian origin. CAD was defined according to strict clinical criteria. Prior to analyses, patients were grouped according to age and gender-specific tertiles of plasma adiponectin levels. Multivariate Cox proportional hazards regression was used to estimate the association between plasma adiponectin levels and age at diagnosis of CAD. Overall, FH patients in the lowest tertile of plasma adiponectin exhibited CAD at a significantly younger age (hazard ratio=1.73, confidence interval 95%: [1.19-2.53]; p=0.004). These results suggest that low plasma adiponectin is associated with an increased risk of premature CAD over and above the already exaggerated risk seen in FH patients. PMID- 17123537 TI - Lack of effect of sugar cane policosanol on plasma cholesterol in Golden Syrian hamsters. AB - Policosanol is a mixture of higher aliphatic alcohols shown to have beneficial effects on plasma lipid levels in animals and humans. Over 50 studies have reported significant reductions in plasma cholesterol using policosanol obtained from Cuban sugar cane (Dalmer, La Havana, Cuba). However, other research groups using policosanol from alternative sources have failed to reproduce the efficacy of these alcohols observed in earlier studies. Therefore, the objective of the present study was to compare the cholesterol-lowering effect of the Dalmer sugar cane policosanol (SCP) product versus an alternative mixture of similar policosanol composition. Forty-eight male Golden Syrian hamsters were randomly assigned to four groups and fed experimental diets ad libitum for a period of 4 weeks: (i) non-cholesterol control, (ii) 0.1% cholesterol control, (iii) 0.1% cholesterol diet supplemented with 275 mg/kg diet of Dalmer Cuban sugar cane policosanol and (iv) 0.1% cholesterol diet supplemented with 275 mg/kg diet of alternative sugar cane policosanol. Hamsters were sacrificed and blood was collected at the end of the feeding period. Body weights and food intakes were similar across study groups. Neither of the two policosanol treatments had any significant effect on plasma lipid levels, as compared to cholesterol control. The outcome of the present study questions the clinical usefulness of policosanol mixtures as cholesterol-lowering nutraceuticals. PMID- 17123538 TI - Laser-induced auto-fluorescence (LIAF) as a method for assessing skin stiffness preceding diabetic ulcer formation. AB - Recurrent foot ulceration is a major cause of morbidity in diabetic patients. Discrepancy between the stiffness of the plantar skin and underlying soft tissues may influence the likelihood of ulceration. Tissue properties change with diabetes primarily due to high blood glucose which promotes intermolecular cross linking of structural proteins thus leading to altered structure and function of these structural fibers. This study utilizes a non-invasive method for indirectly assessing skin tissue in the context of plantar ulcer formation in diabetic patients' feet. Control (C, n=13), and diabetic subjects with a history of ulceration (n=16) were matched based on gender, age (42-81years old) and BMI. Six subjects re-ulcerated (U) during their 1-year follow-up. At every visit, each subject's plantar skin was excited with a weak laser light (337nm) to induce tissue fluorescence at three locations on each foot. The spectral area under the curve (AUC) was calculated after background subtraction and normalization. The mean AUC was significantly higher for diabetics compared to control subjects, (mean AUC: 145.6+/-7.2, C=112.6+/-8.3, respectively, p=0.006). For those who re ulcerated (U, n=6), skin site was not a significant factor, but AUC was diminished at the time of re-ulceration (p<0.05). The alteration of intermolecular bonds in diabetic subjects and thinning of skin prior to ulceration could account for these observations. The decrease in AUC prior to an ulcer formation suggests its potential as a marker of tissue changes, which precede ulceration in the diabetic foot. PMID- 17123539 TI - Fluorescence emission from PAMAM and PPI dendrimers. AB - A strong fluorescence emission from poly(amido amine) (PAMAM) dendrimers with different terminal groups or a poly(propylene imine) (PPI) dendrimer was studied under different conditions by varying experimental parameters such as pH value, aging time, temperature, and concentration. The increase of fluorescence intensity was fast at low pH or high temperature but linear with respect to dendrimer concentration. It was reasonable that the formation of a fluorescence emitting moiety had a close relation to protonated tertiary amine groups in PAMAM or PPI dendrimers. Furthermore, oxidation of the tertiary amines was confirmed to play an important role, which was evidently caused by oxygen in air. The results of fluorescence decay indicated that the deactivation of luminescence was raised with increasing temperature. Dendrimers emitted blue photoluminescence along fiber chain templates on a fluorescent microscope. PMID- 17123540 TI - Modulation of larval nutrition affects midgut neutral lipid storage and temporal pattern of transcription factor expression during mosquito metamorphosis. AB - During holometabolous insect development the critical weight marks a physiological transition after which juvenile hormone (JH) concentration decreases to such a level that a subsequent increase in ecdysone titer will initiate metamorphosis. Starvation experiments indicate that the Aedes aegypti critical weight is achieved by 24 h after the last larval-larval molt. When grown at 24 degrees C with excess food, the time between the critical weight and maximum weight (interval to cessation of growth) is about 24 h and pupation occurs about 24 h after the maximum weight is achieved. Oil Red O staining of 3rd and early 4th instars indicates that the midgut is a neutral lipid storage organ during this period. Coincident with the attainment of the critical weight is the depletion of stored midgut neutral lipid. Application of methoprene to 24 h post molt 4th instars results in renewed midgut storage of neutral lipid suggesting that midgut neutral lipid storage is a JH dependent process. Starvation of 4th instars during the 24 h post-molt period suspends development in a fraction of the larvae, and with the resumption of feeding, development resumes. A regimen of starvation and resumption of feeding of 4th instars suggests that JH concentration decreased over a 40 h period after the resumption of feeding and maximum weight is attained about 48 h after the resumption of feeding. We hypothesize that this results in a shortening of the interval to cessation of growth. Real-time PCR experiments indicate that shortening the interval to cessation of growth compresses the time period during which increases in AHR3, AaEcR-B and AaUSP-a expression occur. PMID- 17123541 TI - Evidence for the location of the allosteric activation switch in the multisubunit phosphorylase kinase complex from mass spectrometric identification of chemically crosslinked peptides. AB - Phosphorylase kinase (PhK), an (alphabetagammadelta)(4) complex, regulates glycogenolysis. Its activity, catalyzed by the gamma subunit, is tightly controlled by phosphorylation and activators acting through allosteric sites on its regulatory alpha, beta and delta subunits. Activation by phosphorylation is predominantly mediated by the regulatory beta subunit, which undergoes a conformational change that is structurally linked with the gamma subunit and that is characterized by the ability of a short chemical crosslinker to form beta-beta dimers. To determine potential regions of interaction of the beta and gamma subunits, we have used chemical crosslinking and two-hybrid screening. The beta and gamma subunits were crosslinked to each other in phosphorylated PhK, and crosslinked peptides from digests were identified by Fourier transform mass spectrometry, beginning with a search engine developed "in house" that generates a hypothetical list of crosslinked peptides. A conjugate between beta and gamma that was verified by MS/MS corresponded to crosslinking between K303 in the C terminal regulatory domain of gamma (gammaCRD) and R18 in the N-terminal regulatory region of beta (beta1-31), which contains the phosphorylatable serines 11 and 26. A synthetic peptide corresponding to residues 1-22 of beta inhibited the crosslinking between beta and gamma, and was itself crosslinked to K303 of gamma. In two-hybrid screening, the beta1-31 region controlled beta subunit self interactions, in that they were favored by truncation of this region or by mutation of the phosphorylatable serines 11 and 26, thus providing structural evidence for a phosphorylation-dependent subunit communication network in the PhK complex involving at least these two regulatory regions of the beta and gamma subunits. The sum of our results considered together with previous findings implicates the gammaCRD as being an allosteric activation switch in PhK that interacts with all three of the enzyme's regulatory subunits and is proximal to the active site cleft. PMID- 17123542 TI - Structural and kinetic studies of induced fit in xylulose kinase from Escherichia coli. AB - The primary metabolic route for D-xylose, the second most abundant sugar in nature, is via the pentose phosphate pathway after a two-step or three-step conversion to xylulose-5-phosphate. Xylulose kinase (XK; EC 2.7.1.17) phosphorylates D-xylulose, the last step in this conversion. The apo and D xylulose-bound crystal structures of Escherichia coli XK have been determined and show a dimer composed of two domains separated by an open cleft. XK dimerization was observed directly by a cryo-EM reconstruction at 36 A resolution. Kinetic studies reveal that XK has a weak substrate-independent MgATP-hydrolyzing activity, and phosphorylates several sugars and polyols with low catalytic efficiency. Binding of pentulose and MgATP to form the reactive ternary complex is strongly synergistic. Although the steady-state kinetic mechanism of XK is formally random, a path is preferred in which D-xylulose binds before MgATP. Modelling of MgATP binding to XK and the accompanying conformational change suggests that sugar binding is accompanied by a dramatic hinge-bending movement that enhances interactions with MgATP, explaining the observed synergism. A catalytic mechanism is proposed and supported by relevant site-directed mutants. PMID- 17123543 TI - Predicting bicycle helmet wearing intentions and behavior among adolescents. AB - INTRODUCTION: Cycling accidents in Australia, especially those resulting in head injuries, are a substantive cause of death and disability; but despite legislation and evidence that helmets reduce the risk of head injury, few adolescents wear them. METHOD: This study employed a revised version of the Theory of Planned Behavior (TPB; [Ajzen, I. (1991). The theory of planned behavior. Organizational Behavior and Human Decision Processes, 50, 179-211]) to investigate the determinants of helmet use among a sample of adolescents. Participants in the initial data collection were 294 high school students in Year 8 and Year 11, with 266 completing a follow-up questionnaire measuring behavior over the previous two weeks. RESULTS: Social norms, perceptions of control, and past behavior significantly predicted intentions to use helmets and perceptions of control and past behavior predicted actual helmet use. CONCLUSIONS: Strengthening the routine of helmet use and building young people's confidence that they can overcome any perceived barriers to helmet use will improve adherence to helmet wearing behavior. PMID- 17123544 TI - The prevention of occupational injuries in two industrial plants using an incident reporting scheme. AB - INTRODUCTION: The aim of this study was to examine whether the introduction of an incident reporting scheme with feedback in two industrial plants had an effect on the number of major incidents. METHOD: An intervention design with measurements before the implementation of the incident reporting scheme and two years later was used to examine the relationship between incident rates, safety climate, the willingness to report incidents and perceived management commitment to safety. RESULTS: The results showed that a successful implementation of an incident reporting scheme was followed by a decline in the incidence of major incidents at a Danish metal plant. A key factor in implementing the scheme was top management commitment, which was lacking at another plant, where the implementation of a similar scheme failed. CONCLUSION: Although the study shows some encouraging results concerning the use of incident reporting schemes to prevent occupational accidents, the possibility to draw causal conclusions is limited in the present study, and further studies are needed before the effectiveness of such schemes can be evaluated with certainty. PMID- 17123545 TI - Production supervisor impacts on subordinates' safety outcomes: an investigation of leader-member exchange and safety communication. AB - PROBLEM: Supervisors are increasingly important to the functioning of manufacturing operations, in large part due to their role as leaders. While supervisors' relations and communication with their subordinates are known to be important in influencing subordinates' behavior, little is known about how these two factors will impact subordinates' safety. This study investigated how much each factor contributes to safety-related outcomes for blue-collar production employees. METHOD: Production employees at five Pennsylvania wood manufacturers completed a survey during their work shift. Five hundred and ninety eight employees provided data on leader-member exchange (LMX), safety communication, and safety-related events. Archival data on OSHA recordables were also obtained from the producers' human resources database. RESULTS: Analyses found that the influence of LMX was greater than that of safety communication in predicting safety-related events. Neither LMX nor safety communication was significantly related to OSHA recordables. Results also demonstrated that employee job satisfaction and demographic variables such as gender and age have safety implications. IMPACT ON INDUSTRY: Results from this study further emphasize the importance of production supervisors and illustrate the potential role of leader member exchange in enhancing workplace safety. Specifically, organizations should foster positive social exchange between their employees and supervisors and enhance the leadership qualities of supervisors to help reduce workplace injuries. PMID- 17123546 TI - Longitudinal examination of underage drinking and subsequent drinking and risky driving. AB - INTRODUCTION: Alcohol use, alcohol misuse, and risky driving from adolescence into young adulthood were compared by drinking onset age. METHODS: Surveys were administered in Grades 5/6, 6/7, 7/8, 10, 12, and at approximately age 23. Participants were placed into Drinking Onset groups based on self-reported alcohol use frequency on the adolescent surveys. Driving records were examined in three age periods: under 21, 21-25, and 26+. RESULTS: The earliest drinking initiators reported higher alcohol use and misuse on each survey, and were more likely to have risky driving offenses before age 21 and to have alcohol driving offenses in all three age periods. DISCUSSION: The earliest drinking initiators engaged in risky drinking behavior and risky driving behavior that was consistently higher than those with later drinking initiation, beginning in adolescence and persisting well into young adulthood. PMID- 17123547 TI - Computational studies for the structure and function of mRPE65. AB - The mRPE65 protein is one form of the RPE65 protein and plays a very important role in the visual cycle. However, its 3D structure and detailed mechanism of function are still unclear because of difficulties with isolation and crystallization. This computational study reports a model for the mRPE65 protein structure derived from a model for sRPE65. The natural substrate for RPE65 has been shown to be a retinyl ester and, by utilizing the Autodock and the Ligplot programs, the interactions between the ester and the protein as well as the effects of several mutations on these interactions are studied. Finally, the position of the binding site is proposed based on an iterative process and the effects of the mutations on the binding site are also discussed. PMID- 17123548 TI - Stress transfer in collagen fibrils reinforcing connective tissues: effects of collagen fibril slenderness and relative stiffness. AB - Unlike engineering fibre composite materials which comprise of fibres that are uniform cylindrical in shape, collagen fibrils reinforcing the proteoglycan-rich (PG) gel in the extra-cellular matrices (ECMs) of connective tissues are taper ended (paraboloidal in shape). In an earlier paper we have discussed how taper of a fibril leads to an axial stress up-take which differs from that of a uniform cylindrical fibre and implications for fibril fracture. The present paper focuses on the influence of fibre aspect ratio, q (slenderness), and Young's modulus (stiffness), relative to that of the gel phase, E(R), on the magnitude of the axial tensile stresses generated within a fibril and wider implications on failure at tissue level. Fibre composite models were evaluated using finite element (FE) and mathematical analyses. When the applied force is low, there is elastic stress transfer between the PG gel and a fibril. FE modelling shows that the stress in a fibril increases with E(R) and q. At higher applied forces, there is plastic stress transfer. Mathematical modelling predicts that the stress in a fibril increases linearly with q. For small q values, fibrils may be regarded as fillers with little ability to provide tensile reinforcement. Large q values lead to high stress in a fibril. Such high stresses are beneficial provided they do not exceed the fracture stress of collagen. Modulus difference regulates the strain energy release density, u, for interfacial rupture; large E(R) not only leads to high stress in a fibril but also insures against interfacial rupture by raising the value of u. PMID- 17123549 TI - Do tiny males grow up? Sperm competition and optimal resource allocation schedule of dwarf males of barnacles. AB - Barnacles, marine crustaceans, have three sexual patterns: simultaneous hermaphroditism, dioecy and androdioecy. In dioecy and androdioecy, large individuals (females and hermaphrodites, respectively) are attached by dwarf males. Depending on species, some dwarf males grow up, others do not in their life time. To investigate which environmental conditions affect growth patterns of dwarf males of barnacles, we investigate the evolutionarily stable life history strategy of dwarf males using Pontryagin's maximum principle. Sperm competition among dwarf males and that among dwarf males and large hermaphrodites is taken into account. Dwarf males grow up in food-rich environments, while they do not grow at all in food-poor environments. ESS of the resource allocation schedule between reproduction and growth follows an "intermediate growth strategy" (simultaneous growth and reproduction) for dioecious species, in which sperm competition is not severe. On the other hand, it approaches "bang-bang control" (switching from allocating all resources toward growth then to reproduction), as sperm competition against surrounding large hermaphrodites becomes severe in androdioecious species. PMID- 17123550 TI - Effects of acute exhaustive physical exercise upon glutamine metabolism of lymphocytes from trained rats. AB - Transitory immunosupression is reported after intense exercise, especially after an increase in training overload and in overtraining. The influence of intense exercise on plasma hormones and glutamine concentration may contribute to this effect. However, the effect of such exercise-induced changes upon lymphocyte and glutamine metabolism is not known. We compared glutamine metabolism in lymphocytes in sedentary (SED) and trained rats. Rats from the moderate group (MOD) swam for 6 weeks, 1 h/day, in water at 32+/-1 degrees C, with a load of 5.5% body weight attached to the tail. Animals from the exhaustive group (EXT) trained like MOD, with training increasing to 3 times 1 h a day during the last week, with 150 min rest between each bout. Animals were killed immediately after the last training bout. We observed reduced concentrations of plasma glucose (p<0.05), glutamine (p<0.05), glutamate (p<0.05) in EXT compared to SED. In MOD, decreases in glutamine (p<0.05) were observed. Analyzing lymphocyte metabolism, we observed an increase in lactate production and glutamine consumption (p<0.05) in MOD (p<0.05) compared to SED and a decrease in glutamine consumption (p<0.05) and aspartate production in EXT. An increase in the proliferative response of lymphocytes in MOD and EXT was also observed when stimulated by ConA and LPS similarly to SED. Acute exercise promoted decreased glutamine plasma concentration and changes in glutamine metabolism that did not impair lymphocyte proliferation in exhaustive trained rats. PMID- 17123551 TI - Prenatal stress on the kinetic properties of Ca2+ and K+ channels in offspring hippocampal CA3 pyramidal neurons. AB - Prenatal stress is known to cause neuronal loss and oxidative damage in the hippocampus of offspring rats. To further understand the mechanisms, the present study was undertaken to investigate the effects of prenatal stress on the kinetic properties of high-voltage-activated (HVA) Ca(2+) and K(+) channels in freshly isolated hippocampal CA3 pyramidal neurons of offspring rats. Pregnant rats in the prenatal stress group were exposed to restraint stress on days 14-20 of pregnancy three times daily for 45 min. The patch clamp technique was employed to record HVA Ca(2+) and K(+) channel currents. Prenatal stress significantly increased HVA Ca(2+) channel disturbance including the maximal average HVA calcium peak current amplitude (-576.52+/-7.03 pA in control group and -702.05+/ 6.82 pA in prenatal stress group, p<0.01), the maximal average HVA Ca(2+) current density (-40.89+/-0.31 pA/pF in control group and -49.44+/-0.37 pA/pF in prenatal stress group, p<0.01), and the maximal average integral current of the HVA Ca(2+) channel (106.81+/-4.20 nA ms in control group and 133.49+/-4.59 nA ms in prenatal stress group, p<0.01). The current-voltage relationship and conductance--voltage relationship of HVA Ca(2+) channels and potassium channels in offspring CA3 neurons were not affected by prenatal stress. These data suggest that exposure of animals to stressful experience during pregnancy can exert effects on calcium ion channels of offspring hippocampal neurons and that the calcium channel disturbance may play a role in prenatal stress-induced neuronal loss and oxidative damage in offspring brain. PMID- 17123552 TI - Correlation between erythrocytes deformability and size: a study using a microchannel based cell analyzer. AB - The deformability of erythrocytes is of great importance for oxygen delivery in the microcirculation [Lipowsky, H.H., 2005. Microvascular rheology and hemodynamics. Microcirculation 12, 5-15]. Aging of erythrocytes is associated with a reduction in deformability and also in size. The present work describes an automated cell analyzer which utilizes a glass microchannel and advanced image processing software. Erythrocytes suspended in a high viscosity medium are filmed flowing through the microchannel. Under these conditions, the cells assume different orientations and undergo varying deformations according to their location in the velocity profile. The cell analyzer enables the measurement of individual erythrocyte velocity, deformability and volume at varying depths within the microchannel. The volume of the cells is calculated based on the experimental data and a fluid mechanics model. The results obtained show that, on average, the deformability of the cells increases with increase in their size. Additionally, the behavior of RBCs in a microchannel is investigated, showing promising diagnostic possibilities. PMID- 17123553 TI - Excitatory amino acid receptor-mediated responses in periaqueductal gray neurons are increased during ethanol withdrawal. AB - The ventrolateral periaqueductal gray (PAG) is critical for propagation in the neuronal network for ethanol withdrawal (ETX) seizures, and ethanol is known to alter glutamate effects. This study evaluated changes in glutamate antagonist effects on PAG neurophysiology in brain slices from rats treated with ethanol in vivo. Spontaneous action potentials were rare in control PAG neurons but common during ETX. Spontaneous excitatory postsynaptic potential (EPSP) frequency was increased during ETX, and an AMPA antagonist, 6,7-dinitroquinoxaline-2,3-dione (DNQX) was more effective in suppressing this activity than an NMDA antagonist, 2 amino-7-phosphonoheptanoate (AP7). EPSPs evoked by stimulation of dorsolateral PAG were decreased by AP7 or DNQX in ETX and control neurons. EPSPs of ETX neurons were significantly less sensitive than controls to blockade by AP7 and DNQX. Paired-pulse facilitation of EPSPs was significantly increased during ETX, but paired-pulse inhibition occurred in controls. Thus, PAG hyperexcitability during ETX results from alterations of both NMDA and AMPA receptor-mediated neurotransmission, which may contribute importantly to ETX seizures. These results differ from previous findings in the seizure-initiating site for ETX seizures, inferior colliculus (IC), where NMDA receptor-mediated mechanisms dominate excitability increases during ETX. This dichotomy may be related to the different role played by IC and PAG in the ETX seizure network. PMID- 17123554 TI - Nitrous oxide (N2O) pre- and postsynaptically attenuates NMDA receptor-mediated neurotransmission in the amygdala. AB - The gaseous anaesthetic N(2)O displays analgesic, anxiolytic, and amnesic properties and has addictive psychedelic effects. N(2)O can further act as a neuroprotective agent, but may also become neurotoxic under certain conditions. Here, we employed whole-cell patch-clamp techniques in acute brain slices, and electrical afferent and infrared-guided laser stimulation to examine how N(2)O (65%) can affect NMDA receptor (NMDAR)-mediated synaptic transmission to principal neurons (PNs) of the adult murine basolateral amygdala (BLA). The BLA plays a critical role in anaesthetic-induced amnesia, the formation of aversive memories, as well as in fear and addictive behaviour. We evoked NMDAR-mediated excitatory postsynaptic currents (NMDAR-EPSCs) in PNs of the BLA (BLA-PNs). We found these currents to be markedly decreased by N(2)O via pre- and postsynaptic actions: Without changing their kinetics and open probability, N(2)O impeded the voltage-dependent channel opening of NMDARs in BLA-PNs and diminished their unitary conductance as estimated by non-stationary fluctuation analysis. In addition, our data speak in favour of a N(2)O-produced reduction in the probability of glutamate release at the synapses generating the NMDAR-EPSCs. It is conceivable that these effects not only contribute to anaesthesia and anxiolysis, but also have bearings on learning and memory as well as excitotoxicity in the amygdala. PMID- 17123555 TI - Ladostigil prevents gliosis, oxidative-nitrative stress and memory deficits induced by intracerebroventricular injection of streptozotocin in rats. AB - Glial activation and oxidative-nitrative stress occur at an early stage in Alzheimer's disease (AD). In a rat model of AD, deficits in cerebral glucose utilization and memory were seen 3-4 weeks after intracerebroventricular (icv) injection of streptozotocin (STZ). This study examined whether icv STZ induced glial activation and oxidative-nitrative stress preceded the memory deficits and whether they could be prevented by ladostigil a novel drug, a cholinesterase and monoamine oxidase inhibitor with neuroprotective activity. One week after STZ injection activated microglia and astrocytes were seen in the cortex, around the cannula penetration area, in the hippocampal CA1 region, corpus callosum, medial and lateral septum. The activated astrocytes showed a significant increase in nitrotyrosine immunoreactivity, a measure of oxidative-nitrative stress. Only 3 weeks later were deficits in episodic (object recognition test) and spatial memory (place recognition) seen in STZ-injected rats. Daily oral administrations of ladostigil (1mg/kg) for 1 week, before and after STZ prevented the glial changes, increase in nitrotyrosine immunoreactivity and memory deficits. Taken together the data support the role of glial activation and oxidative-nitrative stress in discrete brain areas in the aetiology of memory deficits and indicate a potential mechanism for their prevention by drug treatment. PMID- 17123556 TI - Neuroprotective effects of ginsenoside-Rg1 in primary nigral neurons against rotenone toxicity. AB - Ginsenoside-Rg1, the pharmacologically active component isolated from ginseng, demonstrated neuroprotective effects on primary cultured rat nigral neurons against rotenone toxicity. Rotenone, a common household pesticide known for its specific and irreversible mitochondria complex I inhibition, has been suggested to be the causal agent of Parkinson's disease (PD) by inducing degeneration of cells in the substantial nigra. The present study demonstrated that co-treatment of rotenone and Rg1 could reduce rotenone-induced cell death by 58% (SEM=+/-5.60; N=3). Rotenone-induced mitochondria membrane potential (MMP, DeltaPsim) depletion was restored and elevated by at least 38% (SEM=+/-2.15; N=3) by Rg1. In addition, Rg1 prevented cytochrome c release from the mitochrondrial membrane and increased the phosphorylation inhibition of the pro-apoptotic protein Bad through activation of the PI3K/Akt pathway. The protective effects of Rg1 was blocked by glucocorticoid receptor antagonist RU486, indicating that the action of Rg1 is mediated through glucocorticoid receptor (GR). In conclusion, Rg1 inhibits the mitochondrial apoptotic pathway and increases the survival chance of the primary cultured nigral neurons against rotenone toxicity. Thus, Rg1 and its related compounds may be developed as protective agents against neurodegenerative diseases induced by mitochondrial toxins. PMID- 17123557 TI - The cerebellum and emotional experience. AB - While the role of the cerebellum in motor coordination is widely accepted, the notion that it is involved in emotion has only recently gained popularity. To date, functional neuroimaging has not been used in combination with lesion studies to elucidate the role of the cerebellum in the processing of emotional material. We examined six participants with cerebellar stroke and nine age and education matched healthy volunteers. In addition to a complete neuropsychological, neurologic, and psychiatric examination, participants underwent [15O]water positron emission tomography (PET) while responding to emotion-evoking visual stimuli. Cerebellar lesions were associated with reduced pleasant experience in response to happiness-evoking stimuli. Stroke patients reported an unpleasant experience to frightening stimuli similar to healthy controls, yet showed significantly lower activity in the right ventral lateral and left dorsolateral prefrontal cortex, amygdala, thalamus, and retrosplenial cingulate gyrus. Frightening stimuli led to increased activity in the ventral medial prefrontal, anterior cingulate, pulvinar, and insular cortex. This suggests that alternate neural circuitry became responsible for maintaining the evolutionarily critical fear response after cerebellar damage. PMID- 17123558 TI - A vision for public health in the 21st century. PMID- 17123559 TI - An efficient synthesis of 5alpha-androst-1-ene-3,17-dione. AB - 5alpha-Androst-1-ene-3,17-dione (5) as a prodrug of 1-testosterone (4) was prepared in four steps from 17beta-Acetoxy-5alpha-androstan-3-one (stanolone acetate) (1) in high yield. Thus, stanolone acetate (1) was brominated in the presence of hydrogen chloride in acetic acid to give 17beta-acetoxy-2-bromo 5alpha-androstan-3-one (2), which underwent dehydrobromination using lithium carbonate as base with lithium bromide as an additive to give 17beta-acetoxy 5alpha-androst-1-en-3-one (3) in almost quantitative yield with 97% of purity. Compound (3) was hydrolyzed with sodium hydroxide to give 17beta-hydroxy-5alpha androst-1-en-3-one (4,1-testosterone), which was oxidized with chromium trioxide to afford 5alpha-androst-1-ene-3,17-dione (5). The overall yield of 5 was 78.2% with purity of 99%. In this method, the formation of 4-ene was diminished when 1 ene was introduced, and its mechanism was also discussed. PMID- 17123560 TI - Microsatellite evolution: Markov transition functions for a suite of models. AB - This paper takes from the collection of models considered by Whittaker et al. [2003. Likelihood-based estimation of microsatellite mutation rates. Genetics 164, 781-787] derived from direct observation of microsatellite mutation in parent-child pairs and provides analytical expressions for the probability distributions for the change in number of repeats over any given number of generations. The mathematical framework for this analysis is the theory of Markov processes. We find these expressions using two approaches, approximating by circulant matrices and solving a partial differential equation satisfied by the generating function. The impact of the differing choice of models is examined using likelihood estimates for time to most recent common ancestor. The analysis presented here may play a role in elucidating the connections between these two approaches and shows promise in reconciling differences between estimates for mutation rates based on Whittaker's approach and methods based on phylogenetic analyses. PMID- 17123561 TI - Chlorpyrifos, chlorpyrifos-oxon, and diisopropylfluorophosphate inhibit kinesin dependent microtubule motility. AB - Diisopropylfluorophosphate, originally developed as a chemical warfare agent, is structurally similar to nerve agents, and chlorpyrifos has extensive worldwide use as an agricultural pesticide. While inhibition of cholinesterases underlies the acute toxicity of these organophosphates, we previously reported impaired axonal transport in the sciatic nerves from rats treated chronically with subthreshold doses of chlorpyrifos. Those data indicate that chlorpyrifos (and/or its active metabolite, chlorpyrifos-oxon) might directly affect the function of kinesin and/or microtubules--the principal proteins that mediate anterograde axonal transport. The current report describes in vitro assays to assess the concentration-dependent effects of chlorpyrifos (0-10 microM), chlorpyrifos-oxon (0-10 microM), and diisopropylfluorophosphate (0-0.59 nM) on kinesin-dependent microtubule motility. Preincubating bovine brain microtubules with the organophosphates did not alter kinesin-mediated microtubule motility. In contrast, preincubation of bovine brain kinesin with diisopropylfluorophosphate, chlorpyrifos, or chlorpyrifos-oxon produced a concentration-dependent increase in the number of locomoting microtubules that detached from the kinesin-coated glass cover slip. Our data suggest that the organophosphates-chlorpyrifos-oxon, chlorpyrifos, and diisopropylfluorophosphate-directly affect kinesin, thereby disrupting kinesin-dependent transport on microtubules. Kinesin-dependent movement of vesicles, organelles, and other cellular components along microtubules is fundamental to the organization of all eukaryotic cells, especially in neurons where organelles and proteins synthesized in the cell body must move down long axons to pre-synaptic sites in nerve terminals. We postulate that disruption of kinesin-dependent intracellular transport could account for some of the long-term effects of organophosphates on the peripheral and central nervous system. PMID- 17123562 TI - Low level arsenic promotes progressive inflammatory angiogenesis and liver blood vessel remodeling in mice. AB - The vascular effects of arsenic in drinking water are global health concerns contributing to human disease worldwide. Arsenic targets the endothelial cells lining blood vessels, and endothelial cell activation or dysfunction may underlie the pathogenesis of both arsenic-induced vascular diseases and arsenic-enhanced tumorigenesis. The purpose of the current studies was to demonstrate that exposing mice to drinking water containing environmentally relevant levels of arsenic promoted endothelial cell dysfunction and pathologic vascular remodeling. Increased angiogenesis, neovascularization, and inflammatory cell infiltration were observed in Matrigel plugs implanted in C57BL/6 mice following 5-week exposures to 5-500 ppb arsenic [Soucy, N.V., Mayka, D., Klei, L.R., Nemec, A.A., Bauer, J.A., Barchowsky, A., 2005. Neovascularization and angiogenic gene expression following chronic arsenic exposure in mice. Cardiovasc.Toxicol 5, 29 42]. Therefore, functional in vivo effects of arsenic on endothelial cell function and vessel remodeling in an endogenous vascular bed were investigated in the liver. Liver sinusoidal endothelial cells (LSEC) became progressively defenestrated and underwent capillarization to decrease vessel porosity following exposure to 250 ppb arsenic for 2 weeks. Sinusoidal expression of PECAM-1 and laminin-1 proteins, a hallmark of capillarization, was also increased by 2 weeks of exposure. LSEC caveolin-1 protein and caveolae expression were induced after 2 weeks of exposure indicating a compensatory change. Likewise, CD45/CD68-positive inflammatory cells did not accumulate in the livers until after LSEC porosity was decreased, indicating that inflammation is a consequence and not a cause of the arsenic-induced LSEC phenotype. The data demonstrate that the liver vasculature is an early target of pathogenic arsenic effects and that the mouse liver vasculature is a sensitive model for investigating vascular health effects of arsenic. PMID- 17123563 TI - 1,25-Dihydroxyvitamin D3 regulates genes responsible for detoxification in intestine. AB - 1Alpha,25-Dihydroxyvitamin D(3) (1,25-(OH)(2)D(3)), the biologically active form of vitamin D(3), not only plays a major role in mammalian calcium and phosphorous homeostasis but also exerts pleiotropic effects on cell proliferation, differentiation and the immune system. Further, vitamin D is believed to play a significant role in the prevention of colon, prostate, and breast cancer and in reducing the risk of autoimmune diseases. To gain insight into the mechanism whereby vitamin D can have such diverse actions, we have employed microarray technology. We studied the effect of a single dose of 1,25-(OH)(2)D(3) on gene expression in the intestine of vitamin D-deficient rats. Within 6 h, 1,25 (OH)(2)D(3) stimulates the expression of several phase I and phase II biotransformation genes. There is also an increased expression of antioxidant genes. These results support the idea that vitamin D is a significant factor in detoxification and protection against environmental toxins. PMID- 17123564 TI - In vitro analysis of the effects of mutations in the G-tract of the human immunodeficiency virus type 1 polypurine tract on RNase H cleavage specificity. AB - The recognition and precise cleavage of the polypurine tract (PPT) of the human immunodeficiency virus type 1 (HIV-1) is an essential step in HIV-1 reverse transcription. The accurate cleavage, and the subsequent removal, of the PPT by the RNase H activity of HIV-1 RT defines the left end of the double-stranded viral DNA genome, the substrate for integration into the host genome. Previous analyses have shown that mutations in the 3'-end (G-tract) of the PPT cause alterations in RNase H cleavage specificity. In particular, mutations at positions 2 and 5 in the G-tract increased the frequency of retention of PPT sequences in the 2-LTR circle junction. To better understand why these mutations affected PPT cleavage in vivo, we analyzed the cleavage of PPT substrates in vitro that contained altered sequences and unusual base substitutions. Our results, herein, confirm that mutations at positions 2 and 5 of the G-tract do significantly alter the cleavage specificity at the PPT/U3 junction, and further suggest that the miscleavages observed in vivo were due to an improper generation of the PPT primer, as opposed to its improper removal. Finally, our results point to the structure of the PPT, rather than the base-specific contacts between the PPT and HIV-1 RT, as the primary determinants of RNase H cleavage specificity at the PPT/U3 junction. PMID- 17123565 TI - Atomic force microscopy investigation of Mason-Pfizer monkey virus and human immunodeficiency virus type 1 reassembled particles. AB - Particles of DeltaProCANC, a fusion of capsid (CA) and nucleocapsid (NC) protein of Mason-Pfizer monkey virus (M-PMV), which lacks the amino terminal proline, were reassembled in vitro and visualized by atomic force microscopy (AFM). The particles, of 83-84 nm diameter, exhibited ordered domains based on trigonal arrays of prominent rings with center to center distances of 8.7 nm. Imperfect closure of the lattice on the spherical surface was affected by formation of discontinuities. The lattice is consistent only with plane group p3 where one molecule is shared between contiguous rings. There are no pentameric clusters nor evidence that the particles are icosahedral. Tubular structures were also reassembled, in vitro, from two HIV fusion proteins, DeltaProCANC and CANC. The tubes were uniform in diameter, 40 nm, but varied in length to a maximum of 600 nm. They exhibited left handed helical symmetry based on a p6 hexagonal net. The organization of HIV fusion proteins in the tubes is significantly different than for the protein units in the particles of M-PMV DeltaProCANC. PMID- 17123566 TI - Resistance of HIV-1 to the broadly HIV-1-neutralizing, anti-carbohydrate antibody 2G12. AB - The 2G12 mAb inhibits the infection of HIV-1 laboratory-adapted viruses at 50% inhibitory concentrations (IC(50)) ranging from 0.02 to 0.2 microg/ml when evaluated in different cell-types. However, isolates from various HIV-1 subtypes (such as clade C, D, A/E, F and group O) were not inhibited by 2G12 mAb (IC(50) >20 microg/ml). 2G12 mAb pressure in HIV-1 IIIB- and NL4.3-infected T cell cultures selected for resistant viruses containing only few (1 to 3 N glycosylation) deletions in gp120. The 2G12-resistant viruses keep their full sensitivity to various mannose-specific lectins and other known HIV entry inhibitors. Moreover, we observed that the NL4.3-2G12-resistant virus, with the N295K mutation in gp120, became significantly more sensitive to several mannose specific lectins. This is, to our knowledge, the first report showing that a resistant virus generated in vitro against a neutralizing mAb and containing a mutation in gp120, has increased sensitivity to another class of HIV entry inhibitors. PMID- 17123567 TI - Computational prediction and identification of HLA-A2.1-specific Ebola virus CTL epitopes. AB - Ebola virus (EBOV) is known to cause a severe hemorrhagic fever resulting in high mortality. Although the precise host defense mechanism(s) that afford protection against EBOV is not completely understood, T cell-mediated immune responses is believed to play a pivotal role in controlling virus replication and EBOV infection. There have been no reports on mapping of MHC Class I-binding CTL epitopes for EBOV till to date. In this study, we identified five HLA-A2-binding 9-mer peptides of EBOV nucleoprotein (NP) using computer-assisted algorithm. The peptides were synthesized and examined for their ability to bind to MHC class I molecules using a flow cytometry based MHC stabilization assay. Three of the EBOV NP peptides tested (FLSFASLFL, RLMRTNFLI and KLTEAITAA) stabilized HLA-A2. The ability of the HLA-A2-binding EBOV-NP peptides to generate peptide-specific CTLs was evaluated in HLA-A2.1 transgenic mice. Epitope-specific CTL responses were confirmed by cytotoxic assays against peptide-pulsed target cells and interferon gamma ELISPOT assay. Each of the EBOV-NP peptides induced CTL responses in HLA-A2 transgenic mice. Interestingly, all the three peptides were conserved in three different strains of Ebola (Zaire and Reston and Sudan). Taken together, these findings provide direct evidence for the existence of EBOV-derived NP epitopes that may be useful in the development of protective immunogens for this hemorrhagic virus. PMID- 17123568 TI - Labeling HIV-1 virions with two fluorescent proteins allows identification of virions that have productively entered the target cell. AB - GFP-Vpr labeled HIV-1 virions have provided a method to visually examine the interactions between the virus and target cell during infection. However, existing methods to discriminate between virions that have been non-specifically endocytosed from those that have productively entered the host cell cytoplasm have remained problematic. Therefore, we examined the ability of a series of membrane-targeted fluorescent fusion protein constructs to be incorporated into virions. We find that a fluorescent protein fusion targeted to the plasma membrane by the addition of the N-terminal 15 amino acid sequence of c-Src (S15) is efficiently packaged into HIV virions. Using fluorescent proteins fused to this sequence, we have generated virions dually labeled with S15-mCherry and GFP Vpr. Importantly, we can detect the loss of this S15-mCherry membrane signal following fusion. After infection with VSV-g pseudotyped HIV virions, we find a measurable, specific loss of membrane label during infection. This loss of fluorescence is not observed when fusion is prevented using bafilomycin A. This increased ability to discriminate between non-productively endocytosed virions and those actively undergoing steps in the infectious process will facilitate efforts to examine early steps in infection microscopically. PMID- 17123569 TI - Effects of light intensity and water temperature on oxygen release from roots into water lettuce rhizosphere. AB - The oxygen release rate into the rhizosphere by a floating aquatic plant-water lettuce-was determined under various light intensities (0.0-1.2x10(5)lx) and water temperatures (10-35 degrees C). The net specific oxygen release rate was expressed by a model equation comprising the gross oxygen release rate and the rhizosphere respiration terms. Experimental and simulated results show that the net specific oxygen release rate increased with light intensity up to the optimal value, but slight inhibition by higher light intensities was observed at 10-20 degrees C. With increased water temperature, the respiration rate became larger than the gross oxygen release rate. The maximum net specific oxygen release rate of 11.0-12.5mg-O(2)kg-wet(-1)h(-1) was obtained at the optimal condition of about 25 degrees C and 9.0x10(4)-1.1x10(5)lx. The net oxygen release rate was negligible at 35 degrees C at any light intensity because the respiration rate was much greater than the gross oxygen release rate into the rhizosphere. PMID- 17123570 TI - Removal of some organic pollutants in water employing ceramic membranes impregnated with cross-linked silylated dendritic and cyclodextrin polymers. AB - Triethoxysilylated derivatives of poly(propylene imine) dendrimer, polyethylene imine and polyglycerol hyperbranched polymers and beta-cyclodextrin have been synthesized and characterized. These compounds impregnated ceramic membranes made from Al(2)O(3), SiC and TiO(2) and subsequently sol-gel reaction led to their polymerization and chemical bond formation with the ceramic substrates. The resulting organic-inorganic filters were tested for the removal of a variety of organic pollutants from water. They were found to remove of polycyclic aromatic hydrocarbons (up to 99%), of monocyclic aromatic hydrocarbons (up to 93%), trihalogen methanes (up to 81%), pesticides (up to 43%) and methyl-tert-butyl ether (up to 46%). PMID- 17123571 TI - Modeling Cryptosporidium parvum oocyst inactivation and bromate in a flow-through ozone contactor treating natural water. AB - A reactive transport model was developed to simultaneously predict Cryptosporidium parvum oocyst inactivation and bromate formation during ozonation of natural water. A mechanistic model previously established to predict bromate formation in organic-free synthetic waters was coupled with an empirical ozone decay model and a one-dimensional axial dispersion reactor (ADR) model to represent the performance of a lab-scale flow-through ozone bubble-diffuser contactor. Dissolved ozone concentration, bromate concentration (in flow-through experiments only), hydroxyl radical exposure and C. parvum oocyst survival were measured in batch and flow-through experiments performed with filtered Ohio River water. The model successfully represented ozone concentration and C. parvum oocyst survival ratio in the flow-through reactor using parameters independently determined from batch and semi-batch experiments. Discrepancies between model prediction and experimental data for hydroxyl radical concentration and bromate formation were attributed to unaccounted for reactions, particularly those involving natural organic matter, hydrogen peroxide and carbonate radicals. Model simulations including some of these reactions resulted in closer agreement between predictions and experimental observations for bromate formation. PMID- 17123572 TI - Electron transfer from NADH bound to horse liver alcohol dehydrogenase (NAD+ dependent dehydrogenase): visualisation of the activity in the enzyme crystals and adsorption of formazan derivatives by these crystals. AB - The crystals of holoenzyme from native and cross-linked alcohol dehydrogenase exhibit electron transfer from NADH to phenazinium methosulfate (PMS), and then to the tetrazolium salt sodium 3,3'-{1-[(phenylamino)carbonyl]-3,4-tetrazolium} bis(4-methoxy-6-nitro)benzenesulfonate (XXT). The slow dissociation of the cofactor and/or the conformational change associated can now be bypassed. The reduction product, formazan, did not diffuse out of the crystals in buffer and the crystals turned colored. In the presence of dimethyl sulfoxide or dimethoxyethane, the formazan diffused out to the solution. The reaction rates were found to be, respectively, 18% and 15% of the redox reaction rate of ethanol with cinnamaldehyde, close to the activity determined for the enzyme in solution in the presence of dimethoxyethane. The use of system PMS-tetrazolium salt is a useful tool to visualize the activity of dehydrogenases and other electron transferring systems in the crystalline state. The adsorption of formazan by the alcohol dehydrogenase crystals occurs in solution. PMID- 17123573 TI - Weight and height at 4 and 7 years of age in children born to mothers with a high intake of fish contaminated with persistent organochlorine pollutants. AB - In Sweden the main exposure route for persistent organochlorine pollutants (POP) is through consumption of fatty fish from the Baltic Sea (off the eastern coast). The present study aimed to investigate whether intrauterine exposure for POP may have negative impact on children's weight and height at 4 and 7 years of age, respectively. The study included 174 fishermen's wives from the Swedish east coast who had given birth to an infant with either low (n=55) or normal (n=119) birth weight, and 88 and 206 corresponding women from the Swedish west coast (where the fish is less polluted). Comparisons between the east and west coast cohorts were performed. In addition, blood samples were collected among the east coast women and the concentrations of 2,2',4,4',5,5'-hexachlorobiphenyl (CB-153) in plasma was analyzed and estimated for the year of childbirth. There were no significant differences between the east and west coast cohorts regarding weight and height at 4 and 7 years of age. There were, however, significant negative associations between the estimated plasma concentrations of CB-153 during year of childbirth and weight at 4 and 7 years of age, respectively, among the normal birth weight children. The study gives only very weak support for the hypothesized association. PMID- 17123574 TI - Removal of Pb and MDF from contaminated soils by EDTA- and SDS-enhanced washing. AB - Heavy metal- and organic-contaminated sites are ubiquitous, but few studies have been conducted to address such an issue. EDTA- and SDS-enhanced washing was studied for remediation of Pb- and/or marine diesel fuel (MDF)-contaminated soils. The feasibility of recovery and reuse of EDTA and SDS, as well as the physicochemical interactions among the chemical agents, contaminants and soils were extensively investigated using batch experiments. The optimal washing sequence was then determined. The experimental results showed that EDTA could be recovered and reused for four cycles without significant loss of its chelating capacity, while the extraction capability of SDS was noticeably reduced after each reuse cycle. The free phase of marine diesel fuel (MDF) in soils physically isolated the sorbed Pb on soils and thus reducing its extraction by EDTA. The presence of SDS alone or together with low concentration of EDTA was found to enhance Pb removal probably via electrostatic interaction and dissolution of soil organic matter. However, it hindered Pb extraction by high concentration of EDTA, because of the potential formation of complexes between some strongly-bound Pb and SDS, that are more resistant to desorption. Therefore, EDTA washing followed by SDS achieved the highest Pb removal efficiency. On the other hand, MDF removal by SDS was significantly hindered by coexisting Pb in soils, probably because the formation of Pb-dodecyl sulfate (DS) complex would decrease the effective amount of SDS available for forming micelles in solution and enhance MDF sorption. EDTA alone or together with SDS could enhance MDF removal, but the residual MDF after EDTA-washing became more resistant to SDS removal. Consequently, SDS washing followed by EDTA is considered as the optimal washing sequence for MDF removal. PMID- 17123575 TI - Biodegradation of chlordane and hexachlorobenzenes in river sediment. AB - Contamination of river sediments by persistent organic pollutants (POPs) is a worldwide concern, and microbial degradation is regarded as an important process for removal of POPs from river sediments. To date, there is still a lack of systematic study on chlordane biodegradation in river sediments, and the information on hexachlorobenzene (HCB) biodegradation in river sediments is very limited in Japan. We investigated the anaerobic biodegradation potential of trans chlordane (TC), cis-chlordane (CC), and HCB in sediment samples collected at three sites along the Kamogawa River in Saitama Prefecture, Japan. Lag period and biodegradation rates of TC and CC in the three sediments varied greatly with their properties and contamination by TC and CC. In contrast, biodegradation of HCB in all three sediments started immediately with the start of the experiment without lag period, and major differences in biodegradation rates among the sediments were not observed. At the end of 20-week anaerobic incubation in the dark at 30 degrees C temperature, degradation rates ranged from 0.0% to 33.0% for TC, 0.0% to 12.0% for CC, and 47.6% to 59.4% for HCB. Results showed that the high-to-low order of biodegradation in the river sediments was HCB>TC>CC. Although the sediments were collected in the same river, their biodegradation potential varied with properties. Sediment with rich organic content and contamination by TC and CC or HCB was observed to have high biodegradation rates for these pollutants. In addition, biodegradation of TC, CC and HCB was companied by obvious methane generation and drop of oxidation-reduction potential (ORP). PMID- 17123576 TI - Factors affecting accumulation of thallium and other trace elements in two wild Brassicaceae spontaneously growing on soils contaminated by tailings dam waste. AB - Thallium is a scarce, highly toxic element. There are several investigations that report Tl accumulation in plants of the family Brassicaceae. These plants could pose a risk in areas where Tl is present at higher concentrations than normal soils. The present study reports analyses of two wild Brassicaceae, Hirschfeldia incana and Diplotaxis catholica, growing spontaneously at five sampling sites moderately polluted with Tl and other trace elements in the Green Corridor of the Guadiamar river, Seville, S. Spain. In general, trace element content was unremarkable in all part plants, despite the concentrations present in soil. Thallium was the only element whose concentration in both plant species was above normal for plants (maximum values of 5.00 mgkg(-1) in H. incana flowers). There were significant positive correlations between total Tl in soil and Tl in both plant species. Transfer Coefficients (TC) for all elements were, in general, <1 for both species, except for Tl in flowers and fruits at some sites. The highest Enrichment Factor (EF) was found for Tl in H. incana fruits (EF = 607) and D. catholica flowers (EF = 321). H. incana was studied in a previous growing season (2004) in the same area, although the rainfall was 3 times more than in the year of the present study (2005), giving a maximum Tl content of 46.5 mgkg(-1) in H. incana flowers. The data presented here show that Tl content of plants growing in semi-arid conditions can be significantly influenced by precipitation. In dry years, plant Tl accumulation may be significantly reduced. PMID- 17123577 TI - Multicriteria optimisation of a simultaneous supercritical fluid extraction and clean-up procedure for the determination of persistent organohalogenated pollutants in aquaculture samples. AB - A useful tool based on a single-step extraction and clean-up procedure for the determination of 15 organohalogenated pollutants (including brominated flame retardants) in aquaculture samples, using aluminium oxide basic and acidic silica gel in the supercritical extraction cell followed by gas chromatography with electron capture detection or mass spectrometry has been developed. This effective clean-up step ensures a minimum of chromatographic difficulties related to complex matrix components such as aquaculture feed. The extraction procedure has been screened by a fractional factorial design for the preliminary statistically significant parameters. The factors selected were extraction temperature, pressure, static extraction time, dynamic extraction time and carbon dioxide flow rate. The Doehlert design, followed by a multicriteria decision making strategy, was then performed in order to determine the optimum conditions for the two most significant factors: pressure (165 bar) and dynamic extraction time (27 min). Under optimal conditions, the procedure developed with GC-MS/MS provides an excellent linearity, detection (0.01-0.2 ng g(-1)) and quantification limits (0.05-0.8 ng g(-1)) for most of the analytes investigated. The feasibility of the proposed supercritical fluid extraction method was validated by analysing two reference materials and fish feed and shellfish samples with satisfactory results. PMID- 17123578 TI - Biotransformation of flumequine by the fungus Cunninghamella elegans. AB - The metabolism of the antibacterial fluoroquinolone drug flumequine by Cunninghamella elegans was investigated using cultures grown in Sabouraud dextrose broth with 308microM flumequine. The cultures were extracted with ethyl acetate; metabolites were separated by high-performance liquid chromatography and identified by mass spectrometry and proton nuclear magnetic resonance spectroscopy. Flumequine was transformed to two diastereomers of 7 hydroxyflumequine (23 and 43% of the total chromatographic peak area at 280nm) and 7-oxoflumequine (11% of the total peak area). This is the first time that the two 7-hydroxy diastereomers have been characterized structurally; the hydroxyflumequines are known to have less antimicrobial activity than flumequine. PMID- 17123579 TI - Assessment of the natural attenuation of chlorinated ethenes in an anaerobic contaminated aquifer in the Bitterfeld/Wolfen area using stable isotope techniques, microcosm studies and molecular biomarkers. AB - The in situ degradation of chlorinated ethenes was assessed in an anaerobic aquifer using stable isotope fractionation approaches, microcosm studies and taxon specific detection of specific dehalogenating groups of bacteria. The aquifer in the Bitterfeld/Wolfen region in Germany contained all chlorinated ethenes, benzene and toluene as contaminants. The concentrations and isotope composition of the chlorinated ethenes indicated biodegradation of the contaminants. Microcosm studies confirmed the presence of in situ microbial communities capable of the complete dechlorination of tetrachloroethene. Taxon specific investigation of the microbial communities indicated the presence of various potential dechlorinating organisms including Dehalococcoides, Desulfuromonas, Desulfitobacterium and Dehalobacter. The integrated approach, using metabolite spectra, molecular marker analysis and isotope studies, provided several lines of evidence for natural attenuation of the chlorinated ethenes. PMID- 17123580 TI - Effect of heavy metal stress on antioxidative enzymes and lipid peroxidation in leaves and roots of two mangrove plant seedlings (Kandelia candel and Bruguiera gymnorrhiza). AB - The effects of multiple heavy metal stress on the activity of antioxidative enzymes and lipid peroxidation were studied in leaves and roots of two mangrove plants, Kandelia candel and Bruguiera gymnorrhiza, grown under control (10 per thousand NaCl nutrient solution) or five levels of multiple heavy metal stress (10 per thousand NaCl nutrient solution containing different concentration of Pb2+, Cd2+, and Hg2+). Leaves and roots of control and heavy metal-stressed plants were harvested after two months. In leaves of heavy metal-stressed plants superoxide dismutase (SOD) and peroxidase (POD) activities fluctuated in different stress levels compared to the control, while catalase (CAT) activity increased with stress levels in K. candel, but remained unchanged in leaves of B. gymnorrhiza. In comparison with the control, the dynamic tendency of SOD, CAT, and POD activities in roots of heavy metal-stressed plants all ascended, and then declined. The increase in enzyme activities demonstrated that K. candel is more tolerant to heavy metals than B. gymnorrhiza. Lipid peroxidation was enhanced only in leaves of heavy metal-stressed B. gymnorrhiza. These results indicate that in heavy-metal stress antioxidative activities may play an important role in K. candel and B. gymnorrhiza and that cell membrane in leaves and roots of K. candel have greater stability than those of B. gymnorrhiza. For pollution monitoring purposes, POD activity in roots and leaves maybe serve as a biomarker of heavy metal stress in K. candel, while lipid peroxidation maybe serve as biomarker in B. gymnorrhiza. PMID- 17123581 TI - Removal of a broad range of surfactants from municipal wastewater--comparison between membrane bioreactor and conventional activated sludge treatment. AB - Elimination of alkylphenol ethoxylates (APEO) and their degradation products (alkylphenols and alkylphenoxy carboxylates), as well as linear alkylbenzene sulfonates (LAS) and coconut diethanol amides (CDEA), was studied in a pilot plant membrane bioreactor (MBR) working in parallel to a full-scale wastewater treatment plant (WWTP) using conventional activated sludge (CAS). In the CAS system 87% of parent long ethoxy chain NPEOs were eliminated, but their decomposition yielded persistent acidic and neutral metabolites which were poorly removed. The elimination of short ethoxy chain NPEOs (NP(1)EO and NP(2)EO) averaged 50%, whereas nonylphenoxy carboxylates (NPECs) showed an increase in concentrations with respect to the ones measured in influent samples. Nonylphenol (NP) was the only nonylphenolic compound efficiently removed (96%) in the CAS treatment. On the other hand, MBR showed good performance in removing nonylphenolic compounds with an overall elimination of 94% for the total pool of NPEO derived compounds (in comparison of 54%-overall elimination in the CAS). The elimination of individual compounds in the MBR was as follows: 97% for parent, long ethoxy chain NPEOs, 90% for short ethoxy chain NPEOs, 73% for NPECs, and 96% for NP. Consequently, the residual concentrations were in the low mug/l level or below it. LAS and CDEA showed similar elimination in the both wastewater treatment systems that were investigated, and no significant differences were observed between the two treatment processes. Nevertheless, for all studied compounds the MBR effluent concentrations were consistently lower and independent of the influent concentrations. Additionally, MBR effluent quality in terms of chemical oxygen demand (COD), NH(4)(+) concentration and total suspended solids (TSS) was always superior to the ones of the CAS and also independent of the influent quality, which demonstrates high potential of MBRs in the treatment of municipal wastewaters. PMID- 17123582 TI - Sorption of triazoles to soil and iron minerals. AB - Triazoles, additives in runway de-icers, are found in soil and groundwater at airport sites. To better understand the fate and transport of benzotriazole (BTA) and methylbenzotriazole (MeBTA) and to assess possible remediation options of contaminated groundwater, sorption to various soils and ferrous sorbents has been studied. In batch experiments, limited non-linear sorption of BTA to mineral subsoil from the Oslo International Airport, Gardermoen was observed. The sorption to soil could be described using a Freundlich isotherm. pH affected sorption of BTA to subsoil, although the effect was not strong. Increased sorption was observed to zerovalent iron (Fe(0)). MeBTA showed similar sorption behaviour as BTA although the sorption coefficient was generally higher. Sorption to Fe(0) seems to be controlled by multi-layer coverage. Our data suggest that sorption of triazoles to Fe(2)O(3) is negligible. However BTA sorption to 2-line and 6-line ferrihydrites showed strong sorption. The results demonstrate that triazoles are highly mobile in the subsurface environment, however zerovalent iron can be an effective medium for groundwater remediation. Without remediation, wide distribution of triazoles in the environment can be expected due to its extensive application and limited degradability. PMID- 17123583 TI - Sequential treatment via Trametes versicolor and UV/TiO2/Ru(x)Se(y) to reduce contaminants in waste water resulting from the bleaching process during paper production. AB - An efficient sequential, biological and photocatalytic treatment to reduce the pollutant levels in wastewater due to the bleaching process during paper production is reported. For a biological pre-treatment, 800 ml of non-sterilized effluent was inoculated with Trametes versicolor immobilized in polyurethane foam, with 25 g l(-1) glucose, 6.75 mM CuSO(4), and 0.22 mM MnSO(4) added, and cultured at 25 degrees C with an air flow of 800 ml min(-1) for 8d. The fungus did not inhibit growth of the heterotropic populations of the effluent. After 4d of culture, the chemical oxygen demand (COD) reduction and colour removal (CR) were 82% and 80%, respectively, with laccase (LAC) and manganese peroxidase (MnP) activities of 345 U l(-1) and 78 U l(-1), respectively. The COD reduction and CR correlated positively (p<0.0001) with LAC and MnP activities. Chlorophenol removal was 99% of pentachlorophenol, 99% of 2,3,4,6-tetrachlorophenol (2,3,4,6 TCP), 98% of 3,4-dichlorophenol (3,4-DCP) and 77% of 4-chlorophenol (4-CP), while 2,4,5-trichlorophenol (2,4,5-TCP) increased to 0.2 mg l(-1). The pre-treated effluent was then exposed to a photocatalytic treatment. The treatment with photolysis resulted in 9% CR and 46% COD reduction, 42% CR and 60% COD reduction by photocatalysis, and 62% CR and 85% COD reduction by heterogeneous photocatalysis with the system TiO(2)/Ru(x)Se(y) (Fig. 4). With this treatment the bacterial and fungal populations also decreased by 5 logarithmic units with respect to the biological treatment alone (Fig. 5). The total sequential treatment resulted in a 92% CR (from 5800 UC), 97% COD reduction (from 59 g l( 1)) and 99% chlorophenol removal at 96 h and 20 min. PMID- 17123584 TI - Multivariant statistical analysis of PCDD/FS in sewage sludges from different areas of the Valencian Community (Spain). AB - The levels of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in 31 sewage sludges from different wastewater treatment plants corresponding to rural, urban and industrial areas in the Valencian Community (Spain) were analysed. Values of 5.1-346.2 ng I-TEQ kg(-1) (dry weight) were detected for sewage sludge with the highest value in one sample from an industrial area. Therefore the majority of the samples did not exceed the limit proposed (100 ng I TEQ kg(-1)) by the [EU, 2000. Working document on sludge, 3rd draft. Brussels. Available from: http://europa.eu.int/comm/environment/waste/sludge/sludge_en.pdf] for use in agriculture. The dominant congeners for each family of compounds were 1,2,3,4,6,7,8-HpCDD and OCDD from PCDDs, and 1,2,3,4,6,7,8-HpCDF and OCDF from PCDFs. The total concentrations of PCDD/Fs were evaluated statistically through SPPS 11.0 for Windows. The principal component analysis (PCA) was used to extract two PCs as a linear combination of the original variables, one of them associated to urban+highly industrial areas and the other one to urban+low industrial areas. The linear regression method was applied and an efficient correlation was obtained between the total I-TEQ values for each sample and two of most abundant congeners (OCDF and OCDD). This expression was obtained with the results of the 31 samples analysed and a variety of data from other authors. Furthermore, several bilateral correlations between the different congeners completed the statistical analysis. PMID- 17123585 TI - A new Homo erectus molar from Sangiran. PMID- 17123586 TI - Improving cell therapy--experiments using transplanted telomerase-immortalized cells in immunodeficient mice. AB - Cell therapy is the use of stem cells and other types of cells in various therapies for age-related diseases. Two issues that must be addressed before cell therapy could be used routinely in medicine are improved efficacy of the transplanted cells and demonstrated long-term safety. Desirable genetic modifications that could be made to cells to be used for cell therapy include immortalization with human telomerase reverse transcriptase (hTERT). We have used a model for cell therapy in which transplantation of adrenocortical cells restores glucocorticoid and mineralocorticoid hormone levels in adrenalectomized immunodeficient mice. In this model, clones of cells that had been immortalized with hTERT were shown to be able to replace the function of the animals' adrenal glands by forming vascularized tissue structures when cells were transplanted beneath the capsule of the kidney. hTERT-modified cells showed no tendency for neoplastic changes. Moreover, a series of experiments showed that hTERT does not cooperate with known oncoproteins in tumorigenesis either in adrenocortical cells or in human fibroblasts. Nevertheless, hTERT was required for tumorigenesis when cells were implanted subcutaneously rather than in the subrenal capsule space. Changes in gene expression make hTERT-modified cells more robust. Understanding these changes is important so as to be able to separately control immortalization and other desirable properties of cells that could be used in cell therapy. Alternatively, desirable properties of transplants might be provided by co transplanted mesenchymal cells: mesenchymal cell-assisted cell therapy. For both hTERT modification and mesenchymal cell-assisted cell therapy, genomics approaches will be needed to define what genetic modifications are desirable and safe in cells used in cell therapy. PMID- 17123587 TI - Chaperonomics, a new tool to study ageing and associated diseases. AB - The participation of molecular chaperones in the process of senescence and in the mechanisms of age-related diseases is currently under investigation in many laboratories. However, accurate, complete information about the number and diversity of chaperone genes in any given genome is scarce. Consequently, the results of efforts aimed at elucidating the role of chaperones in ageing and disease are often confusing and contradictory. To remedy this situation, we have developed chaperonomics, including means to identify and characterize chaperone genes and their families applicable to humans and model organisms. The problem is difficult because in eukaryotic organisms chaperones have evolved into complex multi-gene families. For instance, the occurrence of multiple paralogs in a single genome makes it difficult to interpret results if consideration is not given to the fact that similar but distinct chaperone genes can be differentially expressed in separate cellular compartments, tissues, and developmental stages. The availability of complete genome sequences allows implementation of chaperonomics with the purpose of understanding the composition of chaperone families in all cell compartments, their evolutionary and functional relations and, ultimately, their role in pathogenesis. Here, we present a series of concatenated, complementary procedures for identifying, characterizing, and classifying chaperone genes in genomes and for elucidating evolutionary relations and structural features useful in predicting functional properties. We illustrate the procedures with applications to the complex family of hsp70 genes and show that the kind of data obtained can provide a solid basis for future research. PMID- 17123588 TI - Proceedings of the 3rd International Conference on Functional Genomics of Ageing, March 29-April 1 2006, Palermo, Sicily, Italy. PMID- 17123589 TI - Long-term mobility of fallout 90Sr in ploughed soil, and 90Sr uptake by wheat grain. AB - In this study, we evaluated the mobility of 90Sr in ploughed upland soil, which affects the residual amount in the soil and plant uptake on the basis of long term monitoring data. Paired samples of soil and wheat grain were taken annually from 1961 to 1995 from 8 agricultural fields in Japan, and the concentrations of exchangeable 90Sr in soil and total 90Sr in wheat grain were determined. The concentration of exchangeable 90Sr in ploughed soil decreased exponentially with time. The environmental factor responsible for the decrease of exchangeable 90Sr in the ploughed layer, lambda(e), was determined from the monitoring data of exchangeable 90Sr in the ploughed soil and the amount of fallout-derived deposition. The lambda(e) was larger from 1970 to 1980 than it was from 1980 to 1995, suggesting that an easily removable fraction of 90Sr in soil was preferentially lost from ploughed soil. Among various soil properties that we investigated, the main factor controlling the long-term mobility of 90Sr from ploughed upland soil and 90Sr uptake by wheat grain was the cation-exchange capacity (CEC) of soil. Our experimental results indicate that the entrapment of 90Sr on a cation-exchange site retards the downward migration and wheat uptake of 90Sr from ploughed soil. The empirical parameters that we obtained based on the long-term observation of a wheat-cultivated upland field in Japan could be used as reference data in order to roughly estimate the mobility of 90Sr in ploughed soil and soil-borne 90Sr transfer to wheat grain in the humid Japanese climate. PMID- 17123590 TI - Association between a functional single nucleotide polymorphism in the MDM2 gene and sporadic endometrial cancer risk. AB - OBJECTIVES: MDM2 is an important negative regulator of the p53 tumor suppressor protein. A naturally occurring T/G single nucleotide polymorphism (SNP) in the MDM2 gene promoter, SNP309, causes an increase in MDM2 protein levels and impairment of p53 tumor suppressor activity. SNP309 occurs at a relatively high frequency in the general population and has been associated with accelerated tumorigenesis in hereditary Li-Fraumeni associated cancers as well as in sporadic soft tissue sarcomas. The objective of this study was to examine the association between SNP309 and sporadic endometrial cancer risk. METHODS: Genomic DNA was isolated from 73 patients with endometrial cancer and 79 healthy, female controls. The MDM2 gene promoter region was amplified by PCR and the SNP309 genotype determined by restriction enzyme digestion of the amplified DNA fragment. Unconditional logistic regression analysis was used to determine the relationship between genotypes and endometrial cancer risk and histopathologic features. RESULTS: The homozygous G/G genotype was found in 25% of endometrial cancer cases and 11% of controls. In an age-adjusted analysis of cases and controls, the G/G genotype increased the risk of endometrial cancer 2.76-fold (95% CI: 1.06, 7.20; p=0.03) compared to presence of a wild-type T allele (T/G and T/T genotypes). No association was found between the SNP309 G/G genotype and either endometrial cancer histology, grade, stage, or age at diagnosis. CONCLUSIONS: The MDM2 SNP309 homozygous G/G genotype may be a genetic variant that influences sporadic endometrial cancer susceptibility. PMID- 17123591 TI - Developmental origin of a bipotential myocardial and smooth muscle cell precursor in the mammalian heart. AB - Despite recent advances in delineating the mechanisms involved in cardiogenesis, cellular lineage specification remains incompletely understood. To explore the relationship between developmental fate and potential, we isolated a cardiac specific Nkx2.5(+) cell population from the developing mouse embryo. The majority of these cells differentiated into cardiomyocytes and conduction system cells. Some, surprisingly, adopted a smooth muscle fate. To address the clonal origin of these lineages, we isolated Nkx2.5(+) cells from in vitro differentiated murine embryonic stem cells and found approximately 28% of these cells expressed c-kit. These c-kit(+) cells possessed the capacity for long-term in vitro expansion and differentiation into both cardiomyocytes and smooth muscle cells from a single cell. We confirmed these findings by isolating c-kit(+)Nkx2.5(+) cells from mouse embryos and demonstrated their capacity for bipotential differentiation in vivo. Taken together, these results support the existence of a common precursor for cardiovascular lineages in the mammalian heart. PMID- 17123592 TI - Multipotent embryonic isl1+ progenitor cells lead to cardiac, smooth muscle, and endothelial cell diversification. AB - Cardiogenesis requires the generation of endothelial, cardiac, and smooth muscle cells, thought to arise from distinct embryonic precursors. We use genetic fate mapping studies to document that isl1(+) precursors from the second heart field can generate each of these diverse cardiovascular cell types in vivo. Utilizing embryonic stem (ES) cells, we clonally amplified a cellular hierarchy of isl1(+) cardiovascular progenitors, which resemble the developmental precursors in the embryonic heart. The transcriptional signature of isl1(+)/Nkx2.5(+)/flk1(+) defines a multipotent cardiovascular progenitor, which can give rise to cells of all three lineages. These studies document a developmental paradigm for cardiogenesis, where muscle and endothelial lineage diversification arises from a single cell-level decision of a multipotent isl1(+) cardiovascular progenitor cell (MICP). The discovery of ES cell-derived MICPs suggests a strategy for cardiovascular tissue regeneration via their isolation, renewal, and directed differentiation into specific mature cardiac, pacemaker, smooth muscle, and endothelial cell types. PMID- 17123593 TI - Lycopene protects against cyclosporine A-induced testicular toxicity in rats. AB - Cyclosporine A (CsA)-induced direct failures in hypothalamic-pituitary-gonadal axis and Sertoli cell phagocytic function have been considered for testicular toxicity so far. It has clearly been reported that oxidative stress leads to damage in sperm functions and structure of the testis. Therefore, this study was conducted to demonstrate whether CsA causes testicular and spermatozoal toxicity associated with the oxidative stress, and to investigate the possible protective effect of lycopene against CsA-induced damages in all reproductive organs and sperm characteristics in male rats. While the daily administration of CsA at the dose 15 mg/kg for 21 days significantly decreased the seminal vesicles weight, epididymal sperm concentration, motility, testicular tissue glutathione (GSH), glutathione peroxidase (GSH-Px) and catalase (CAT), diameter of seminiferous tubules and germinal cell thickness, it increased malondialdehyde (MDA) level and abnormal sperm rates along with degeneration, necrosis, desquamative germ cells in testicular tissue. However, the CsA along with simultaneous administration of lycopene at the dose of 10mg/kg markedly ameliorated the CsA-induced all the negative changes observed in the testicular tissue, sperm parameters and oxidant/antioxidant balance. In conclusion, CsA-induced oxidative stress leads to the structural and functional damages in the testicular tissue and sperm quality of rats and, lycopene has a potential protective effect on these damages. PMID- 17123594 TI - Reduced sensitivity to prosodic attitudes in adults with focal right hemisphere brain damage. AB - Although there is a strong link between the right hemisphere and understanding emotional prosody in speech, there are few data on how the right hemisphere is implicated for understanding the emotive "attitudes" of a speaker from prosody. This report describes two experiments which compared how listeners with and without focal right hemisphere damage (RHD) rate speaker attitudes of "confidence" and "politeness" which are signalled in large part by prosodic features of an utterance. The RHD listeners displayed abnormal sensitivity to both the expressed confidence and politeness of speakers, underscoring a major role for the right hemisphere in the processing of emotions and speaker attitudes from prosody, although the source of these deficits may sometimes vary. PMID- 17123595 TI - Shear versus micro-shear bond strength test: a finite element stress analysis. AB - OBJECTIVES: This study aimed at comparing the stress distribution in shear and micro-shear test set-ups using finite element analysis, and suggesting some parameter standardization that might have important influence on the results. METHODS: Two-dimensional plane strain finite element analysis was performed using MSCPatran and MSCMarc softwares. Model configurations were based on published experimental shear and micro-shear test set-ups and material properties were assumed to be isotropic, homogeneous and linear-elastic. Typical values of elastic modulus and Poisson's ratios were assigned to composite, dentin and adhesive. Loading conditions considered a single-node concentrated load at different distances from the dentin-adhesive interface, and proportional geometry (1:5 scale, but fixed adhesive layer thickness in 50microm) with similar calculated nominal strength. The maximum tensile and shear stresses, and stress distribution along dentin-adhesive interfacial nodes were analyzed. RESULTS: Stress distribution was always non-uniform and greatly differed between shear and micro-shear models. A pronounced stress concentration was observed at the interfacial edges due to the geometric change: stress values farther exceeded the nominal strength and tensile stresses were much higher than shear stresses. For micro-shear test, the relatively thicker adhesive layer and use of low modulus composites may lead to relevant stress intensification. An appropriate loading distance was established for each test (1mm for shear and 0.1mm for micro-shear) in which stress concentration would be minimal, and should be standardized for experimental assays. SIGNIFICANCE: The elastic modulus of bonded composites, relative adhesive layer thickness and load application distance are important parameters to be standardized, once they influence stress concentration. PMID- 17123596 TI - Analysis of ligand binding curves on basis of mean intrinsic thermodynamic quantities. AB - The mean intrinsic thermodynamic quantity can be defined by considering the relative population of complex species in the solution and the value of intrinsic thermodynamic quantity corresponds to each step of ligation. In the present study a new method is introduced for analysis of experimental ligand binding data on basis of mean intrinsic thermodynamic quantities. In this regard, a deviation parameter was defined by comparing the non-interacting system with the cooperative interactive one. This parameter can be calculated just by estimation of the first binding constant. A set of relations between this deviation parameter and other binding characteristics, such as mean intrinsic Gibbs free energy of binding and mean Gibbs free energy of site-site interaction, have been developed. This model presents binding mechanism in a unified way that is simple, yet stringent, more straightforward, more reliable and informative. This analyzing method has been successfully applied for evaluation of various systems such as oxygen binding to hemoglobin, laurate and warfarin binding to human serum albumin, and reveals some new biological features of these binding systems. PMID- 17123597 TI - Modelling of fracture behaviour in biomaterials. AB - One of the most frequent causes of degradation and failure of quasi-brittle biomaterials is fracture. Mechanical breakdown, even when not catastrophic, is of particular importance in the area of biomaterials, as there are many clinical situations where it opens the path for biologically mediated failures. Over the past few decades the materials/biomaterials community has developed a number of numerical models, but only with limited incorporation of brittle failure phenomena. This article investigates the ability of a non-linear elastic fracture mechanics (NLEFM) model to reliably predict failure of biomaterials with a specific focus on the clinical settings of restorative dentistry. The approach enables one to predict fracture initiation and propagation in a complex biomechanical status based on the intrinsic material properties of the components. In this paper, we consider five examples illustrating the versatility of the present approach, which range from the failure of natural biomaterials, namely dentine and enamel, to a restored tooth, a three unit all ceramic bridge structure and contact-induced damage in the restorative layered materials systems. It is anticipated that this approach will have ramifications not only to model fracture events but also for the design and optimisation of the mechanical properties of biomaterials for specific clinically determined requirements. PMID- 17123598 TI - Physico-chemical-mechanical and in vitro biological properties of calcium phosphate cements with doped amorphous calcium phosphates. AB - Calcium phosphate cements (CPCs) are successfully used as bone substitutes in dentistry and orthopaedic applications. This study investigated the physico chemical-mechanical properties of and in vitro biological properties (cell response) of CPCs prepared with amorphous calcium carbonate phosphate (ACCP) doped with magnesium (ACCP-Mg), zinc (ACCp-Zn) or fluoride (ACCP-F) ions. The experimental CPC consisted of alpha-TCP, doped ACCP, and MPCM powders as matrix and biphasic calcium phosphate (BCP) granules. X-ray diffraction analysis showed that the matrix converted to apatite with poor crystallinity (reflecting small crystal size) after setting for 24 h, while BCP remained apparently unchanged. Cements with ACCP-F (F-CPC) had shorter setting times and greater compressive strength compared to cements with ACCP-Mg (Mg-CPC) or ACCP-Zn (Zn-CPC). Scanning electron microscopy (SEM) showed that crystals set on Mg-CPC and Zn-CPC were smaller compared to those on F-CPC. The total porosity of Mg-CPC was greater compared to Zn-CPC or F-CPC. Osteoblast-like cells, MC3T3-E1, remained viable and maintained their ability to express alkaline phosphatase in contact with the CPCs with doped ACCPs. PMID- 17123599 TI - Coupled substitution of type A and B carbonate in sodium-bearing apatite. AB - A suite of Na-bearing type A-B carbonate hydroxyapatites {Ca(10-y)Na(y)[(PO4)(6 y)(CO3)y][(OH)(2-2x)(CO3)x], x approximately = y} has been synthesized at 1200 degrees C and 0.5-1.0 GPa, and investigated by single-crystal X-ray structure and FTIR spectroscopy. Crystal data for the maximum content of carbonate (11.1 wt%) are a = 9.3855(7), c = 6.9142(4) A, space group P6(3)/m, R = 0.023, R(w) = 0.014. Structural accommodation of the substitutions requires local coupling of Na and channel (type A) and phosphate (type B) carbonate ion defects. The type B carbonate ion is located on the sloping faces of the substituted phosphate group, but is inclined at an angle of 53 degrees to the mirror plane. FTIR spectra have minimal nu3 absorption beyond 1500 cm(-1) and dominant nu2 absorption at 873 cm( 1). Synthetic Na-bearing type A-B apatites (with a high content of type A carbonate) are thus similar in both chemical composition and infrared spectra to biological apatites. The latter are reinterpreted as Na-bearing type A-B carbonate apatites with channel carbonate up to 50% of total carbonate. PMID- 17123600 TI - Nanostructured calcium phosphates (NanoCaPs) for non-viral gene delivery: influence of the synthesis parameters on transfection efficiency. AB - Calcium phosphate (CaP) based approaches remain an attractive option for delivering plasmid DNA (pDNA) into cultured cells. However, despite their appeal, current synthesis methodologies typically yield lower, less consistent transfection efficiencies when compared to viral approaches. Therefore, we report here a novel method to consistently synthesize efficient, nano-sized, mono dispersed CaP-pDNA particles; accomplished by optimizing both the stoichiometry (Ca/P ratio) of the CaP particles as well as the mode in which the calcium and phosphate precursor solutions are mixed. Our results indicate that calcium and phosphate precursors when mixed in a controlled and regulated manner reproducibly result in nano-sized particles that consistently yield higher transfection efficiencies when compared to particles synthesized via manual mixing (a two-fold increase was observed). Also, maximum transfection efficiencies in both HeLa and MC3T3-E1 cells lines were obtained when a Ca/P ratio between 100 and 300 was used. Particles synthesized within this optimum Ca/P ratio range were between 25 and 50 nm. Our data suggests that these maximized transfection efficiencies were obtained because these particles not only effectively condensed (70% efficient) but also efficiently bound (90% efficient) the pDNA. In addition, X-ray diffraction and Fourier transform infrared spectroscopy analyses confirmed that all of the synthesized CaP structures exhibited the hydroxyapatite phase. PMID- 17123601 TI - Culturing of glial and neuronal cells on polysialic acid. AB - Although peripheral nerves exhibit regeneration capacities after transection injuries, the success of nerve repair depends crucially on the length of the gap. In addition to autologous nerve grafting as the conventional neurosurgical treatment to overcome long gaps, alternative strategies are needed. Numerous experimental studies have been undertaken to find the optimal material for production of artificial prostheses, which can be introduced as conduits between the nerve stumps. The current study follows the aim to establish polysialic acid (polySia), a homopolymer of alpha2,8-linked sialic acid residues, as a novel, biocompatible, and bioresorbable material for nerve tissue engineering. As a first step towards this goal, protocols for efficient coating of cell culture dishes with soluble polySia were established. In addition, primary nerve cells which are candidates for reconstructive therapies, including neonatal and adult Schwann cells, neural progenitor cells, spinal ganglionic neurons and motoneurons were cultured on polySia substrates. Cultures were evaluated with regard to cell survival and cell proliferation capacities. polySia turned out to be stable under cell culture conditions, and induced degradable and degradation products had no negative effects on cell cultures. Furthermore, polySia revealed its compatibility for several cell types derived from rat embryonic, postnatal and adult nervous tissue when used as a substrate. PMID- 17123602 TI - Inhibition of in vitro chondrogenesis in RGD-modified three-dimensional alginate gels. AB - The goal of this study was to investigate the effects of adhesion to the arginine glycine-aspartic acid (RGD) sequence on the chondrogenesis of bone marrow stromal cells (BMSCs). Synthetic RGE- and RGD-containing peptides were conjugated to sodium alginate, and bovine BMSCs were seeded onto 2D alginate surfaces or encapsulated in 3D gels. BMSCs spread specifically on RGD-modified surfaces, and spreading was inhibited by a soluble RGD peptide and by anti-beta1 and anti alpha(v)beta3 integrin blocking antibodies. After 7 days in 3D gel culture, the chondrogenic supplements (TGF-beta1 and dexamethasone) significantly stimulated chondrocytic gene expression (collagen II, aggrecan, and Sox-9) and matrix accumulation (collagen II and sGAG) in RGE-modified gels, but this response was inhibited in the RGD-modified gels. Inhibition of sGAG synthesis increased with increasing RGD density, and synthesis was partially rescued by adding a soluble RGD peptide. Addition of an anti-alpha(v)beta3 integrin blocking antibody had no effect on chondrogenesis, while an anti-alpha5 antibody reduced sGAG accumulation. Overall, this study demonstrates that interaction with the RGD motif significantly inhibits the initial chondrogenesis of BMSCs within 3D alginate gels. These results provide new insights into the role of cell-matrix interactions in regulating chondrogenesis and highlight the importance of choosing appropriate biomaterials for tissue engineering therapies. PMID- 17123603 TI - Clinical significance of a blood eosinophilia in adult T-cell leukemia/lymphoma: a blood eosinophilia is a significant unfavorable prognostic factor. AB - We investigated the clinical significance of a blood eosinophilia in a cohort of 158 consecutive patients with adult T-cell leukemia/lymphoma (ATLL), and multivariate analysis revealed that a blood eosinophilia was an independent and a significant unfavorable prognostic factor. Interestingly, a blood eosinophilia was independent of serum LDH level, which might reflect the tumor burden. The present study shows that measurement of the blood eosinophil count is useful for predicting the prognosis and for determining a suitable treatment strategy for ATLL patients. PMID- 17123604 TI - Chemokine system and tissue infiltration in aggressive NK-cell leukemia. AB - NK cell-type lymphoproliferative disease of granular lymphocytes can be subdivided into aggressive NK-cell leukemia (ANKL) and chronic NK-cell lymphocytosis (CNKL). Hepatosplenomegaly is observed in ANKL patients, and hepatic failure is a common cause of death. Significant numbers of ANKL cells were pathologically observed in sinusoidal and interlobular regions of the liver, and in the splenic red pulp. In our previous study, ANKL cells were simultaneously positive for CXCR1 and CCR5. So, in order to elucidate the mechanism in the systemic migration of ANKL cells, we investigated the expression of the corresponding chemokines in ANKL compared with CNKL. The serum level of IL 8, MIP-1alpha and MIP-1beta was significantly elevated in ANKL patients, and ANKL cells were highly positive for IL-8, RANTES, MIP-1alpha and MIP-1beta according to intracellular staining and RT-PCR. These chemokines were also positively stained in hepatocytes. The interaction between Fas and Fas ligand (FasL) is supposed to be one of the mechanisms for liver dysfunction in ANKL. The serum concentration of soluble FasL was significantly high in ANKL patients, and ANKL cells expressed FasL protein in the cytoplasm. These results suggest that the chemokine system plays an important role in the transmigration of FasL-expressing ANKL cells. PMID- 17123605 TI - The discovery process: what mothers see and do in gaining awareness of the sexual abuse of their children. AB - OBJECTIVE: The aim of this study was to explore how mothers discovered that their children had been sexually abused. The exploration included learning from whom or in what ways mothers learned about the abuse, whether there were prior suspicions, if actions were taken to determine likelihood of abuse, and the barriers to recognizing abuse. METHOD: An exploratory survey of 125 non-abusive mothers of sexually abused children in three clinical sites was used. The sample included primarily Caucasians and African-Americans in a Midwestern state. A focus group study was used to develop the instrument. The survey was analyzed using descriptive statistics. RESULTS: Mothers first came to learn of sexual abuse from a verbal report (42%) or behaviors (15%) of their victimized child. Almost half of the mothers had a sense that something was "not quite right" prior to knowing about the abuse. Mothers took many actions to try to clarify what was occurring including talking with their child (66%) or watching things more closely (39%). Evidence most convincing mothers of the abuse included child's disclosure (74%), child's behavior (66%), and child's emotions (60%). Factors increasing uncertainty included denial by the abuser (21%). CONCLUSIONS: This exploratory study provides initial data on how mothers come to learn of and believe the sexual abuse of their children. Educating mothers about effective ways to explore suspicions and weigh the evidence for or against abuse may enhance maternal protection and expedite investigations. PMID- 17123606 TI - HLA genes in Cubans and the detection of Amerindian alleles. AB - Caribbean Islands including Cuba were first inhabited by Meso-American and later by Arawak-speaking Amerindians from nowadays Venezuela. Spanish invaders brought to almost extinction to the Amerindian population after 1492. Black slaves from West Africa were taken into Cuba by Europeans. The degree of admixture among populations is approached. HLA alleles were studied by DNA techniques. Comparison with other worldwide populations (a total of 14.094 chromosomes) included genetic distances, Neighbour-Joining dendrograms, correspondence analyses and calculation of extended haplotypes. While African-European HLA features were clearly found, Amerindian HLA characteristics are less evident, indicating that Amerindian devastation was particularly marked after 1492 AD. However, typical Amerindian alleles have been found in our Cuban sample, i.e. DRB1*0403, DRB1*0404, DRB1*0407, DRB1*0411, DRB1*0802 and DRB1*1602. The presence of Amerindian alleles in Cubans [corrected] may have a bear in the making up of transplantation registries (both for bone marrow and solid organ transplantation) at the regional level and also be important for epidemiological studies of diseases linked to HLA. PMID- 17123607 TI - Treatment of acute retinal necrosis syndrome with oral antiviral medications. AB - OBJECTIVE: Acute retinal necrosis (ARN) is a distinct ocular viral syndrome traditionally treated with intravenous acyclovir followed by oral acyclovir. We investigated the use of the oral antiviral medications valacyclovir and famciclovir as the sole treatment for patients with newly diagnosed ARN syndrome. DESIGN: Retrospective, uncontrolled, interventional case series. PARTICIPANTS: Eight consecutive patients with newly diagnosed ARN treated solely with oral antiviral medications. INTERVENTION: All patients received famciclovir or valacyclovir without antecedent intravenous therapy. One patient with bilateral ARN treated with famciclovir received a single intravitreal injection of foscarnet in the more severely involved eye. MAIN OUTCOME MEASURES: Clinically and photographically documented complete resolution of retinitis and best corrected visual acuity on final follow-up. RESULTS: Active retinitis resolved completely in 10/10 (100%) affected eyes. Initial response to treatment was seen as early as 4 days (in 5 eyes), with a median time to complete resolution of 14 days. At the last examination, visual acuity was improved (> or = 2 Snellen lines) in 6 (60%) eyes, stable in 2 (20%) eyes, and worse in 2 (20%) eyes. Over a mean follow-up of 36 weeks (range, 7-72 weeks), 3 eyes developed rhegmatogenous retinal detachment that was successfully repaired with 1 vitrectomy surgery. No patient with initially unilateral involvement developed disease in the contralateral eye. CONCLUSIONS: In this pilot study, the use of the oral drugs valacyclovir and famciclovir resulted in complete regression of herpetic necrotizing retinitis. Additional studies are necessary to evaluate the role of these antiherpetic medications in the treatment of the ARN syndrome. PMID- 17123608 TI - Intraocular lens exchange due to incorrect lens power. AB - PURPOSE: To evaluate patients who had intraocular lens (IOL) exchange for unexpected postoperative refractive errors, determine the sources associated with the errors, and derive an empiric approach to estimating the power for IOL exchange. DESIGN: Retrospective review of interventional case series. PARTICIPANTS: Twenty-two eyes that underwent IOL exchange for correcting unexpected refractive errors after cataract surgery with IOL implantation were reviewed. INTERVENTION: All the IOLs for IOL exchange were placed in-the-bag. The same type of IOLs was used for original IOL implantation and IOL exchange in 91% (20/22) of eyes. MAIN OUTCOME MEASURES: Uncorrected visual acuity, best spectacle corrected visual acuity (BSCVA), refraction, and reasons for IOL exchange. The equation derived from refractive change and change in IOL power was developed to calculate IOL power for exchange. RESULTS: Of the 22 cases, the identified reasons were keratometry errors in 5 (23%) and incorrect axial length (AL) determination in 3 (14%). In 3 other cases, a wrong IOL was implanted. After IOL exchange, 82% (18/22) of eyes were within +/-0.50 diopters (D) and 86% (19/22) within +/-1.00 D of emmetropia. Uncorrected visual acuity was 20/40 or better in 82% of eyes, and BSCVA was 20/40 or better in 95% (21/22) of eyes. The correlation between change of refraction and IOL power was significant (P<0.002). CONCLUSIONS: Among the identified causes, incorrect corneal power determination was the most frequent reason for incorrect IOL power implantation, followed by error in AL measurement and inserting a wrong IOL. The pre-exchange refraction can be used theoretically to calculate the IOL power for exchange. PMID- 17123609 TI - Evaluation of changes in peripapillary nerve fiber layer thickness after deep sclerectomy with optical coherence tomography. AB - PURPOSE: To detect and quantify changes, using optical coherence tomography (OCT), in the peripapillary retinal nerve fiber layer (RNFL) thickness in patients with glaucoma who underwent deep sclerectomy. DESIGN: Prospective, controlled, interventional case series. PARTICIPANTS: Thirty-four consecutive patients who underwent monocular deep sclerectomy (surgery group) and medically treated fellow eyes (control group). METHODS: Quantitative analysis of the peripapillary RNFL by OCT and automated perimetry were performed before surgery and 6 months after surgery in both eyes. MAIN OUTCOME MEASURES: The changes in RNFL thickness overall and by quadrant were evaluated and studied with respect to age, preoperative visual field test global indices, postoperative changes in intraocular pressure (IOP), and postoperative changes in visual field global indices. Changes observed in RNFL thickness were compared between eyes after surgery and in fellow eyes. RESULTS: The IOP decreased from a baseline mean of 23.6+/-5.1 mmHg to 11.7+/-2.9 mmHg (P<0.001) 6 months after surgery at the time of OCT testing. The mean percent IOP change was 48.4+/-15.7%. No significant changes in the mean RNFL thickness overall or by quadrant were observed after surgery or in the mean deviation (MD) and pattern standard deviation after surgery. There was no significant difference in the RNFL thickness between eyes in the surgery group and those in the control group. The mean preoperative visual field MD was significantly (P = 0.006) worse in eyes with a postoperative decrease in the overall RNFL thickness compared with those with an increase in the RNFL thickness. No correlation was found between RNFL thickness changes and age or changes in the visual field global indices. There was no significant difference between eyes with an IOP reduction of more than 50% and those with a reduction in IOP less than 30% (P = 0.514). CONCLUSIONS: The authors found no significant changes in the peripapillary RNFL thickness measured 6 months after deep sclerectomy. The only significant factor related to RNFL thickness changes after surgery was the preoperative visual field MD (P = 0.038). PMID- 17123610 TI - Laser peripheral iridotomy in primary angle-closure suspects: biometric and gonioscopic outcomes: the Liwan Eye Study. AB - PURPOSE: To assess the immediate effect of laser peripheral iridotomy (LPI) and mechanisms of angle closure in a population-based study of primary angle closure (PAC) suspects. DESIGN: Prospective interventional study. PARTICIPANTS: People identified as PAC suspects aged 50 to 79 years from a population-based survey in Guangzhou, China. INTERVENTION: Laser peripheral iridotomy was performed in 1 randomly selected eye. Examinations were carried out before and 2 weeks after the intervention. MAIN OUTCOME MEASURES: Intraocular pressure (IOP), ultrasound biometry, optical pachymetry, and gonioscopy. RESULTS: A total of 72 people with bilateral suspected PAC participated in the study. Mean IOP decreased by 3 mmHg (P<0.001), but axial anterior chamber depth did not change significantly (P = 0.784) after LPI. Median limbal anterior chamber depth increased from 15% to 25% of peripheral corneal thickness (P<0.001, Wilcoxon signed-rank test). Median iridotrabecular angle width increased from 0 degrees to 10 degrees in the superior quadrant and from 10 degrees to 30 degrees in the inferior quadrant (P<0.001). Nevertheless, 14 eyes (19.4%) still had 3 or more quadrants in which the posterior (usually pigmented) trabecular meshwork could not be seen after laser iridotomy. CONCLUSIONS: This study confirms that LPI results in a significant increase in the angle width in Chinese people with narrow angles. However, one fifth of eyes had residual angle closure after LPI. Although this report confirms that iridotomy widens the anterior chamber angle in most PAC suspects, long-term prospective studies with a larger sample size are required to determine if the risks of PAC glaucoma and other related pathologic sequelae are reduced after prophylactic LPI and to investigate the risk-to-benefit ratio before recommending widespread use of prophylactic LPI in this population. PMID- 17123611 TI - Severe reactive ischemic posterior segment inflammation in acanthamoeba keratitis: a new potentially blinding syndrome. AB - OBJECTIVE: To describe a newly recognized clinical syndrome in Acanthamoeba keratitis consisting of severe reactive ischemic posterior segment vascular inflammation. DESIGN: Noncomparative, retrospective, single-institution observational case series. PARTICIPANTS: Five eyes of 5 patients with Acanthamoeba keratitis. METHODS: A retrospective review of the records of patients diagnosed with Acanthamoeba keratitis between January 1, 1995, and December 1, 2005, was conducted to identify those who underwent eventual enucleation. Five enucleated eyes of 118 eyes with Acanthamoeba keratitis were identified. MAIN OUTCOME MEASURES: History, clinical examination results, available laboratory study results, and histopathologic examination results. RESULTS: Histopathologic examination showed Acanthamoeba cysts in the cornea in 4 eyes, whereas it failed to demonstrate amebic cysts or trophozoites in the posterior segment of all eyes studied and unexpectedly revealed chronic chorioretinal inflammation with perivascular lymphocytic infiltration and diffuse neuroretinal ischemia in 4 of 5 eyes. Retinal artery thrombosis was present in 3 of the 4 involved eyes, and central retinal artery and vein thrombosis was found in 1 eye. Hematologic studies in 3 patients showed abnormal anticardiolipin antibody levels in 1 patient and factor V Leiden deficiency in another. CONCLUSIONS: Prolonged Acanthamoeba keratitis can result in a severe sterile ischemic posterior segment inflammation that is potentially blinding, especially in patients with underlying hypercoagulation disorders. PMID- 17123612 TI - Identification of retinal breaks using subretinal trypan blue injection. AB - PURPOSE: To describe the use of subretinal trypan blue to identify retinal breaks during vitrectomy for rhegmatogenous retinal detachment (RD). DESIGN: Interventional case series. PARTICIPANTS: Five patients with RD in whom no retinal break could be identified by internal search with scleral indentation. METHODS: Trypan blue 0.15% was injected transretinally into the subretinal space using a 41-gauge cannula designed for macular translocation surgery. Perfluorocarbon heavy liquid was then injected into the vitreous cavity and the eye was rotated such that trypan blue was vented out of the break. The plume of trypan blue was used to identify retinal breaks, or in some cases staining of the break facilitated break detection. Subretinal fluid was then drained through the break or a drainage retinotomy and surgery was completed using standard techniques. MAIN OUTCOME MEASURE: Identification of previously unseen retinal breaks. RESULTS: This technique successfully identified a retinal break in 4 out of 5 patients. After absorption of the gas tamponade all retinas remained attached with a median visual acuity of 6/12. CONCLUSION: Failure to identify a retinal break during RD surgery is a well-recognized clinical challenge that may adversely affect outcome. In this setting, chromophore-assisted retinal break detection may be a useful surgical technique. PMID- 17123613 TI - High myopia and glaucoma susceptibility the Beijing Eye Study. AB - OBJECTIVE: To evaluate whether marked myopia, compared with moderate myopia and low myopia, is associated with a higher prevalence of glaucomatous optic nerve damage. DESIGN: Population-based cross-sectional study. PARTICIPANTS: Four thousand four hundred thirty-nine of 5324 subjects 40 years or older were invited to participate (response rate, 83.4%). The group was stratified according to refractive error into high myopia (myopia > -8 diopters [D]), marked myopia (<-6 to -8 D), moderate myopia (<-3 to -6 D), low myopia (<-0.5 to -3 D), emmetropia ( 0.5 to + <2 D), and hyperopia (>+ 2 D) subgroups. METHODS: Morphologic assessment of optic disc monoscopic photographs. MAIN OUTCOME MEASURES: Morphologic optic disc parameters and intraocular pressure (IOP). RESULTS: For 4319 (97.3%) subjects (8484 eyes), optic disc photographs were evaluated. Prevalence of glaucomatous optic nerve atrophy as defined by the glaucomatous optic nerve head appearance did not vary significantly (P = 0.77; odds ratio [OR], 1.2; 95% confidence interval [CI], 0.38-3.81) between the highly myopic group and the group with marked myopia. In both refractive groups combined, glaucoma frequency seemed to be higher (P = 0.075; OR, 2.28; 95% CI, 0.99-5.25) higher than in the group with moderate myopia; it was significantly (P = 0.001; OR, 3.5; 95% CI, 1.71-7.25) higher than in the group with low myopia; significantly (P<0.001; OR, 7.56; 95% CI, 3.98-14.35) higher than in the group with emmetropia; and significantly (P = 0.005; OR, 4.23; 95% CI, 1.57-11.45) higher than in the group with hyperopia. Glaucoma frequency did not vary significantly between the hyperopic group and the emmetropic group (P = 0.17), the group with low myopia (P = 0.83), and the group with moderate myopia (P = 0.32). Intraocular pressure did not vary significantly (P>0.10) between any of the subgroups. Similar results were obtained for the frequency of glaucoma defined as glaucomatous optic disc appearance and visual field defects. In binary logistic regression analysis, presence of glaucoma was significantly associated with the myopic refractive error (P<0.001), age (P<0.001), and IOP (P<0.001). CONCLUSIONS: Marked to high myopia with a myopic refractive error exceeding -6 D may be a risk factor associated with glaucomatous optic neuropathy. PMID- 17123614 TI - Hookworm infestation as a risk factor for Mooren's ulcer in South India. AB - OBJECTIVE: To investigate the association between Mooren's ulcer and intestinal hookworm infestation in South India. DESIGN: Prospective observational case control study. PARTICIPANTS: Fifteen patients with Mooren's ulcer and 30 age- and gender-matched controls seen at Aravind Eye Hospital. METHODS: Stool samples from the Mooren's ulcer patients and controls were collected and analyzed for presence of hookworm infestation. MAIN OUTCOME MEASURE: Prevalence of hookworm infestation in Mooren's ulcer patients and controls. RESULTS: There was a statistically significant correlation between intestinal hookworm infestation and the occurrence of Mooren's ulcer (P = 0.009). Retrospective exploratory subgroup analyses suggested that the correlation between intestinal hookworm infestation and the occurrence of Mooren's ulcer in men (P<0.0001) was stronger than the correlation in women, with no statistically significant difference being observed in the prevalence of hookworm infestation between women with Mooren's ulcer and female control subjects (P>0.99). Similarly, when both the Mooren's ulcer and the control subject groups were analyzed retrospectively by age > 50 years or age < 50, subjects with an age over 50 demonstrated a stronger correlation between hookworm infestation and Mooren's ulcer than controls (P = 0.017), whereas there was no statistically significant difference in the prevalence of hookworm infection between Mooren's ulcer subjects and control subjects < or = 50 (P = 0.31). CONCLUSION: Intestinal hookworm infestation appears to be associated with the development of Mooren's ulcer in South India. Although the power of our retrospective exploratory subgroup analyses was limited by multiple testing and small sample sizes, these data suggest further that the correlation between intestinal hookworm infestation and the development of Mooren's ulcer may be greatest in male patients with more advanced age. PMID- 17123615 TI - Relationship between optical coherence tomography-measured central retinal thickness and visual acuity in diabetic macular edema. AB - OBJECTIVE: To compare optical coherence tomography (OCT)-measured retinal thickness and visual acuity in eyes with diabetic macular edema (DME) both before and after macular laser photocoagulation. DESIGN: Cross-sectional and longitudinal study. PARTICIPANTS: Two hundred ten patients (251 eyes) with DME enrolled in a randomized clinical trial of laser techniques. METHODS: Retinal thickness was measured with OCT and visual acuity was measured with the electronic Early Treatment of Diabetic Retinopathy procedure. MAIN OUTCOME MEASURES: Optical coherence tomography-measured center point thickness and visual acuity. RESULTS: The correlation coefficients for visual acuity versus OCT center point thickness were 0.52 at baseline and 0.49, 0.36, and 0.38 at 3.5, 8, and 12 months after laser photocoagulation. The slope of the best fit line to the baseline data was approximately 4.4 letters (95% confidence interval, 3.5-5.3) of better of visual acuity for every 100-mum decrease in center point thickness at baseline with no important difference at follow-up visits. Approximately one third of the variation in visual acuity could be predicted by a linear regression model that incorporated OCT center point thickness, age, hemoglobin A1C, and severity of fluorescein leakage. The correlation between change in visual acuity and change in OCT center point thickening 3.5 months after laser treatment was 0.44, with no important difference at the other follow-up times. A subset of eyes showed paradoxical improvements in visual acuity with increased center point thickening (7%-17% at the 3 time points) or paradoxical worsening of visual acuity with a decrease in center point thickening (18%-26% at the 3 time points). CONCLUSIONS: There is modest correlation between OCT-measured center point thickness and visual acuity, and modest correlation of changes in retinal thickening and visual acuity after focal laser treatment for DME. However, a wide range of visual acuity may be observed for a given degree of retinal edema. Thus, although OCT measurements of retinal thickness represent an important tool in clinical evaluation, they cannot substitute reliably as a surrogate for visual acuity at a given point in time. This study does not address whether short-term changes on OCT are predictive of long-term effects on visual acuity. PMID- 17123616 TI - Sino-orbital fistula: a complication of exenteration. AB - PURPOSE: To report the incidence, characteristics, and management of sino-orbital fistulas, a complication of orbital exenteration. DESIGN: Retrospective interventional case series. PARTICIPANTS: One hundred ten patients who underwent orbital exenteration at 2 orbital units. METHODS: Retrospective chart review of all cases of orbital exenteration between 1993 and 2005 at one orbital unit and between 1999 and 2005 at a second orbital unit. MAIN OUTCOME MEASURES: Incidence of sino-orbital fistulas. RESULTS: Seventy-three and 37 orbital exenterations were performed at the first and second orbital units, respectively. Five patients developed sino-orbital fistulas, 1 of whom developed 2 fistulas at separate sites. In the first unit, 4 fistulas developed in 3 of 73 (4.1%) patients who underwent orbital exenteration. In the second unit, 2 fistulas developed in 2 of 37 (5.4%) exenterated orbits. The majority (5/6) of fistulas occurred medially to the ethmoid sinus, whereas 1 occurred superiorly to the frontal sinus. Risk factors that may have contributed to fistula formation include radiotherapy (3/6), sinus disease (3/6), intraoperative penetration into a sinus (3/6), and immunocompromise (1/6). Management was tailored to the individual case and ranged from conservative socket hygiene to surgical repair with grafts or flaps. Four of the 6 fistulas recurred after repair. Three of these subsequently were closed successfully. Only 1 fistula persisted until the patient died from malignant disease. CONCLUSIONS: Sino-orbital fistulas are uncommon but not rare complications of orbital exenteration that may be predicted by several risk factors. Bothersome symptoms may necessitate treatment, which can range from conservative management to surgical repair with various grafts or flaps. Despite repair, fistulas may be difficult to eradicate. PMID- 17123617 TI - Nonprogressive glaucomatous cupping and visual field abnormalities in young Chinese males. AB - OBJECTIVE: To describe a series of young to middle-aged men of Chinese origin who presented with a constellation of ocular findings suggestive of glaucoma, that were found to be stable over a 7-year period. DESIGN: Retrospective case series. PARTICIPANTS: Sixteen 25- to 66-year-old male patients. METHODS: Medical records of the participants, of Chinese origin and referred for glaucoma evaluation over a 7-year period, were reviewed. All patients underwent complete ophthalmic examinations, stereo imaging of the optic nerves, and automated perimetry. Fewer than 5% of all patients seen in this practice were of Chinese origin. The patients were observed for the duration of the study in a single glaucoma clinic. MAIN OUTCOME MEASURES: Visual field (VF) changes or progressive optic nerve cupping suggestive of glaucoma. RESULTS: The patients had an average age of 38.9 years. Tilted discs were present in 75.0% (24/32) and peripapillary atrophy in 81.3% (26/32) of nerves. Cup-to-disc ratios ranged from 0.20 to 0.95 and averaged 0.56. The lowest intraocular pressure (IOP) in any patient at any time was 8 mmHg, whereas the highest was 29 (average range, 13.5-17.9). Intraocular pressure lowering therapy had been used in 56.3% (9/16). There was a family history of presumed glaucoma in 25.0% (4/16) of patients. High myopia (>-6.00-diopter spherical equivalent [SE]) was present in 43.8% of eyes (14/32), and SEs ranged from -11.25 to +0.25. The most common VF defect was an arcuate defect, found in 31.3% (10/32) of patients. There were no females of Chinese origin with similar findings identified during this period. Neither optic nerve nor significant VF progression was found during the follow-up period, regardless of the use of IOP lowering therapy. CONCLUSIONS: These young Chinese patients previously diagnosed with glaucoma or considered glaucoma suspects had stable ocular findings for up to 7 years, irrespective of IOP-lowering therapy. Their condition was associated with myopia and tilted discs. Many were being treated with IOP-lowering therapy for glaucoma, a condition they may not have had. Further prospective epidemiologic study is needed to determine whether such a constellation of nonprogressive findings is more common in young Chinese males than in the general population. PMID- 17123618 TI - Clinical characteristics of Vogt-Koyanagi-Harada syndrome in Chinese patients. AB - PURPOSE: To characterize the clinical features of Vogt-Koyanagi-Harada syndrome (VKH) in Chinese patients. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: A total of 410 consecutive VKH patients examined from August 1995 to April 2005. METHODS: Charts of these patients were reviewed according to international VKH criteria. Patients who consulted us within 2 weeks after a uveitis attack were classified into group 1; between 2 weeks and 2 months into group 2; and after 2 months into group 3. The history and clinical findings of all of the patients were reviewed. Laser flare-cell photometry, fundus fluorescein angiography, indocyanine green angiography, optical coherence tomography, B-scan ultrasonography, and ultrasound biomicroscopy were performed in certain cases. Corticosteroids were mainly used to treat patients in groups 1 and 2, whereas cyclosporine or chlorambucil in combination with corticosteroids were prescribed for patients in group 3. MAIN OUTCOME MEASURES: Demographics, clinical presentation, and ancillary examinations of the patients in all 3 groups. RESULTS: The patients were diagnosed as having either complete (n = 273), incomplete (n = 76), or probable (n = 61) VKH syndrome. Exudative retinal detachment and either choroiditis or chorioretinitis were the main findings in group 1. Posterior uveitis with mild to moderate anterior uveitis simulating a nongranulomatous inflammation as evidenced by dust keratic precipitates, anterior chamber cells, and flare was noted in group 2. Recurrent granulomatous anterior uveitis with a "sunset glow" fundus was observed in group 3. Complicated cataract was the most common complication in the patients in group 3. At the final visit, the uveitis was completely controlled in all patients and a best-corrected visual acuity of <20/200 was 1.9%/eye-year, 1.2%/eye-year, and 6%/eye-year in groups 1, 2, and 3, respectively. CONCLUSION: Vogt-Koyanagi-Harada syndrome in Chinese patients is characterized by early posterior uveitis, and if the syndrome is not controlled, subsequent recurrent granulomatous anterior uveitis. Good visual prognosis is possible if the patients are managed according to a tailored immunosuppressive treatment protocol. PMID- 17123619 TI - Alignment artifacts in optical coherence tomography analyzed images. AB - PURPOSE: To compare retinal thickness and retinal height of the original scanned optical coherence tomography (OCT) images with those of the same images after automated retinal thickness analysis. DESIGN: Cross-sectional study. PARTICIPANTS: Thirty normal eyes, 20 eyes with neovascular age-related macular degeneration (AMD), 20 with central serous chorioretinopathy (CSC), 20 with macular holes, and 20 with non-AMD related macular edema from 110 subjects were selected randomly from an OCT database. One of the 6 macular scans in each eye was chosen randomly for analysis. METHODS: Two sets of OCT images--original images and analyzed images (after retinal thickness [single eye] analysis)--in each eye were exported for retinal thickness or retinal height measurement. Comparisons of retinal thickness or retinal height at selected locations were performed. MAIN OUTCOME MEASURES: Retinal thickness (defined as the distance between the anterior boundary of the retinal nerve fiber layer [RNFL] and posterior boundary of the photoreceptor layer) and retinal height (defined as the distance between the anterior boundary of the RNFL and baseline level of the anterior boundary of the retinal pigment epithelium [RPE]). Retinal height was measured when there was detachment of neurosensory retina or RPE. RESULTS: No significant difference in retinal thickness was observed between the original and the analyzed OCT images in normal eyes and in eyes with macular holes or non-AMD related macular edema. However, OCT-analyzed images demonstrated retinal thickness or retinal height measurements in eyes with CSC or neovascular AMD significantly lower than the corresponding measurements in the original images (all with Ps< or =0.001, Wilcoxon signed rank test). In the groups of neovascular AMD and CSC, Bland-Altman plots revealed mean differences of 124 mum (95% limits of agreement between -65.5 and 313.6) and 84.4 mum (95% limits of agreement between -178.0 and 346.8), respectively, between the original and analyzed retinal measurements. CONCLUSIONS: Retinal thickness and retinal height could be underestimated in patients with CSC or neovascular AMD after retinal thickness analysis in Stratus OCT when either automatic measurements or manual caliper assisted measurements are performed on the analyzed images. We recommend exporting the original scanned OCT images for retinal thickness and retinal height measurement in patients with CSC or neovascular AMD. PMID- 17123620 TI - Differences in clinical findings between Caucasians and African Americans with biopsy-proven sarcoidosis. AB - OBJECTIVE: To study the prevalence of ocular manifestations in African American and Caucasian patients with biopsy-proven sarcoidosis at the initial ophthalmic examination and to determine the relationship between angiotensin-converting enzyme (ACE) levels, chest x-ray findings, and ocular signs of sarcoidosis. DESIGN: Retrospective, cross-sectional, observational study. PARTICIPANTS: Eighty one consecutive patients with biopsy-proven sarcoidosis seen at the Doheny Eye Institute from January 1989 through April 2005. METHODS: Medical records were reviewed to obtain demographic data, biopsy site, initial ocular findings, pulmonary symptoms, and results of serum ACE levels and chest x-rays. Associations between ACE level/chest x-ray stages and ocular manifestations related to sarcoidosis were obtained from these data. MAIN OUTCOME MEASURES: Ocular manifestations related to sarcoidosis. RESULTS: Of the 81 patients, 35 were Caucasian; 29 were African American; and the remaining 17 were Hispanic, Asian Indian, and other races. Female patients were older than males (P = 0.05). Sixty-five (80%) of the 81 patients had ocular manifestations related to sarcoidosis. Thirty-three patients (40.7%) had uveitis, 12 (14.8%) had adnexal granulomas, and 25 (30.8%) had keratoconjunctivitis sicca. Of the 33 patients with uveitis, 22 presented with nongranulomatous inflammation. There was no significant association between ocular manifestations related to sarcoidosis and serum ACE levels (P = 0.43) or chest x-ray stage (P>0.99). Of the 29 African American patients, 26 (89.7%) had ocular manifestations related to sarcoidosis, compared with 24 (68.6%) of the 35 Caucasians (P = 0.12). The African American patients were younger (mean age, 44.4 years) than the Caucasian patients (mean age, 52.0) (P = 0.003) and had higher mean ACE levels (P = 0.003). A significantly high proportion of African American males presented with uveitis (P = 0.005), and a significantly high proportion of African American females presented with adnexal granulomas (P = 0.05). CONCLUSIONS: The present study reveals that patients with sarcoidosis can present initially with clinical features of nongranulomatous uveitis. Relative to Caucasians, African American patients with sarcoidosis tend to be younger when they first present to the ophthalmologist and to present with uveitis and/or adnexal granuloma. Serum ACE levels and chest x-ray stages may not help predict the occurrence of ocular changes in sarcoidosis. PMID- 17123621 TI - A clinical and optical coherence tomography study of the margins of choroidal colobomas. AB - OBJECTIVE: To describe the features at the margin of the choroidal colobomas as evaluated clinically and by optical coherence tomography (OCT). DESIGN: Prospective observational case series. PARTICIPANTS: Seventeen patients (30 eyes) that presented to the outpatient department of a tertiary care center. TESTING: Clinical examination and OCT testing of the 30 eyes. MAIN OUTCOME MEASURES: Description of OCT features of the margin of the choroidal colobomas. RESULTS: Histologically, there is no normal choroid, retinal pigment epithelium, or retina overlying choroidal colobomas; rather, the overlying tissue is an extension of the retina called the intercalary membrane (ICM). In these patients, OCT showed that transition from normal retina to the ICM could be categorized as abrupt or gradual and also showed that the inner neurosensory retinal layers continued as the ICM, whereas the outer layers could not be traced beyond a point. In some cases with apparently attached retina, subclinical retinal detachments were identified along the margin of the coloboma. In cases with retinal detachment, OCT allowed for identification of the precise site of communication between the sub-ICM space and subretinal space at the locus minoris resistentiae. The margin of the choroidal coloboma, in some cases, showed the appearance of a hump owing to inward turning of the retinochoroidal layers with thickening of the layers at the margin. In small colobomas, OCT revealed the ICM thickness comparable to normal retina but showed structural alterations. Fundus lesions that were clinically labeled forme fruste of choroidal coloboma seemed to have normal retinal thickness and layering on OCT. In 1 patient, a temporal optic pit was associated with the forme fruste choroidal coloboma in 1 eye; the fellow eye had a typical choroidal coloboma. CONCLUSIONS: Optical coherence tomographic evaluation of the margins of choroidal colobomas helps in understanding the transition from retina to ICM, detects subclinical retinal detachments, and aids in identifying the site of communication between the sub-ICM space and the subretinal space in eyes with retinal detachment. PMID- 17123622 TI - Refractive error and visual impairment in school children in rural southern China. AB - PURPOSE: To assess the prevalence of refractive error and visual impairment in school children in a rural area of southern China. DESIGN: Prospective cross sectional survey. PARTICIPANTS: Two thousand four hundred children from junior high schools in Yangxi County. METHODS: Random selection of classes from the 3 junior high school grade levels was used to identify the study sample. Children from 36 classes in 13 schools were examined in April 2005. The examination included visual acuity (VA) testing; ocular motility evaluation; cycloplegic autorefraction; and examination of the external eye, anterior segment, media, and fundus. MAIN OUTCOME MEASURES: Distance VA and cycloplegic refraction. RESULTS: Among 2515 enumerated children, 2454 (97.6%) were examined. The study population consisted of the 2400 children between 13 and 17 years old. Prevalences of uncorrected, presenting, and best-corrected VA < or = 20/40 in the better eye were 27.0%, 16.6%, and 0.46%, respectively. Sixty percent of those who could achieve acuity > or =20/32 in at least one eye with best correction were without the necessary spectacles. Refractive error was the cause in 97.1% of eyes with reduced vision; amblyopia, 0.81%; other causes, 0.67%; and unexplained causes, 1.4%. Myopia (spherical equivalent, -0.50 diopters [D] or more in either eye) affected 36.8% of 13-year-olds, increasing to 53.9% of 17-year-olds. Myopia was associated with higher grade level, female gender, schooling in the county urban center, and higher parental education. Hyperopia (+2.00 D or more) affected approximately 1.0% in all age groups. Astigmatism (> or =0.75 D) was present in 25.3% of all children. CONCLUSIONS: Reduced vision because of uncorrected myopia is a public health problem among school-age children in rural China. Effective VA screening strategies are needed to eliminate this easily treated cause of visual impairment. PMID- 17123623 TI - Comparison of visual field defects using matrix perimetry and standard achromatic perimetry. AB - PURPOSE: To compare visual field (VF) defects found by Swedish interactive thresholding Algorithm (SITA) perimetry and Matrix perimetry, a new VF device that utilizes frequency doubling technology in a 24-2 test pattern. DESIGN: Prospective cross-sectional study. PARTICIPANTS: Fifty eyes from 50 subjects with SITA field defects were recruited for an observational study. METHODS: Swedish Interactive Threshold Algorithm and Matrix VF testing were performed on patients from a glaucoma practice. To evaluate the learning effect on the performance of the VF, we tested subsets of each group who had previous experience with standard automated perimetry (SAP). MAIN OUTCOME MEASURES: Test duration, mean threshold, mean deviation (MD), pattern standard deviation (PSD), glaucoma hemifield test, and number of abnormal points on the pattern deviation plot were evaluated for each device. RESULTS: Test duration was significantly shorter for Matrix (SITA, 357.0+/-85.6 seconds; Matrix, 319.5+/-16.5 seconds; P = 0.0002, paired t-test). Thirty-six percent of eyes with SITA VF defects showed a normal Matrix field. In 30 of 32 eyes (94%) where both devices showed VF defects, the defects were congruent. Mean threshold value was significantly lower with Matrix compared to SITA (P<0.0001, paired t-test), as was MD (-5.34+/-5.42 dB, -4.14+/-5.29 dB, respectively; P = 0.03, paired t-test). There was no significant difference in PSD between the 2 devices (P = 0.78, paired t-test). Matrix delineated significantly smaller (P = 0.005, Wilcoxon's test) and deeper (P<0.001, Wilcoxon's test) defects than those found with SITA. Similar results were observed in the subgroups with prior SAP experience. CONCLUSIONS: The Matrix examination did not detect 36% of abnormal SITA fields. Matrix field defects were smaller and deeper than those appearing in SITA perimetry. PMID- 17123624 TI - Effect of cytomegalovirus retinitis on the risk of visual acuity loss among patients with AIDS. AB - PURPOSE: To describe the prevalence and incidence of reduced visual acuity in eyes of patients with AIDS and without cytomegalovirus (CMV) retinitis at enrollment and estimate the proportion of incident vision loss attributable to new-onset CMV retinitis in this cohort. DESIGN: Multicenter prospective observational study. PARTICIPANTS: Three thousand fourteen eyes of 1507 patients with AIDS and without CMV retinitis at enrollment. METHODS: Medical history, ophthalmologic examination, and laboratory testing collected at enrollment and at follow-up visits every 6 months thereafter. MAIN OUTCOME MEASURES: Loss of visual acuity across the < or =20/50 and < or =20/200 thresholds and doubling of the visual angle; potential causes of this vision loss. RESULTS: For eyes of patients without CMV retinitis at enrollment, the proportions with best-corrected visual acuity of < or =20/50 and of < or =20/200 were 3.9% and 1.8%, respectively. The incidence rates of vision loss to < or =20/50, < or =20/200, and to a doubling of the visual angle were 1.5/100 eye-year (EY), 0.8/100 EY, and 2.1/100 EY, respectively. Approximately 40% of the incident vision loss was attributable to CMV retinitis diagnosed during the follow-up period, and approximately 25% was attributable to cataract. CONCLUSIONS: Although the development of CMV retinitis was the most common reason for visual acuity loss in eyes of our patients with AIDS, it accounted for less than half of the vision loss in our population (approximately 40%). Newly diagnosed cataract during the follow-up period accounted for a substantial amount of incident vision loss as well. PMID- 17123625 TI - Thyrotropin-releasing hormone and its receptors--a hypothesis for binding and receptor activation. AB - Thyrotropin-releasing hormone (TRH), a tripeptide, exerts its biological effects through stimulation of cell-surface receptors, TRH-R, belonging to the superfamily of G protein-coupled receptors (GPCR). Because of the intermediate size of TRH, it is smaller than polypeptide ligands that interact at GPCR ectodomains and larger than biogenic amines, which interact within GPCR transmembrane domains (TMD), the TRH/TRH-R complex probably shares properties of these 2 extremes, representing a unique system to study GPCR/ligand interactions. In this review, we summarize the current knowledge of the structure-activity relationships in the TRH/TRH-R system. Based on experimental data and the structural information acquired from computer simulations, we formulate a working hypothesis to describe the molecular events underlying the processes of TRH binding and TRH-R activation. This hypothesis represents a starting point for understanding the biology of the TRH/TRH-R system on a molecular level and provides a basis for potential design of new potent and selective modulators of TRH-R's activity. PMID- 17123626 TI - Experimental characterization and modelling of analytical monolithic column. AB - Hydrodynamics, equilibrium and kinetics of adsorption in a silica-based monolithic column Chromolith Performance RP-18e (Merck KgaA, Germany) have been studied. The column permeability was calculated according to the Darcy law for laminar flow. The efficiency of the monolithic column was characterized through the height equivalent to a theoretical plate (HETP) for myoglobin, phenol and progesterone. The 2-D single channel mathematical model has been applied to describe the adsorption dynamics. Parabolic velocity profile, axial and radial diffusion in the monolith channel, linear driving force model for the mass transfer in the monolith channel skeleton wall and linear adsorption equilibrium were assumed. The mathematical model gives good prediction of the experimental elution peaks. PMID- 17123627 TI - An inexpensive sub-millisecond system for walking measurements of small animals based on optical computer mouse technology. AB - Stimuli from a broad spectrum of sensory modalities, including visual, auditory, thermal, and chemical, can elicit walking responses in animals, reflecting neural activity in sensorimotor pathways. We have developed an integrated walking measurement system with sub-millisecond temporal accuracy capable of detecting position changes on the order of 100 microm. This tracking system provides the experimenter with a means by which to map out the response spectrum of a tethered animal to any number of sensory inputs on time scales relevant to propagation in the nervous system. The data acquisition system consists of a modified optical computer mouse, a microcontroller with peripheral support circuitry, a binary stimulus sync line, and a serial (RS-232) data transfer interface. The entire system is constructed of relatively inexpensive components mostly converted from commercially available peripheral devices. We have acquired walking data synchronized with auditory stimuli at rates in excess of 2100 samples per second while applying this system to the walking phonotactic response of the parasitic fly Ormia ochracea. PMID- 17123628 TI - Application of simple photobleaching microscopy techniques for the determination of the balance between anterograde and retrograde axonal transport. AB - The directionality of axonal transport represents an important question in neurophysiological and neuropathological research. Various approaches such as videomicroscopy, radioisotopic and fluorescence-based techniques are used. Recently, a novel FRAP-based (fluorescent recovery after photobleaching) technique using synaptophysin-EGFP expression in primary neurons was applied, allowing reliable and sensitive evaluation of gross axonal transport changes using confocal live-imaging microscopy. Here, we describe a novel FLIP-based (fluorescence loss in photobleaching) approach using a synaptophysin-EGFP probe that allows the differential evaluation of the ante- and retrograde transport parameters. Furthermore, we improved the sensitivity of the probe by substituting EGFP with an ECFP/VenusYFP fusion FRET (fluorescence resonance energy transfer) pair. The use of this FRET couple improves the precision of axonal transport measurements by combining FLIP and FLAP (fluorescence localization after photobleaching) techniques and eliminating the need for pre-bleaching images and thus pixel shifts between various exposures, and by reducing the deleterious effect of photobleaching. PMID- 17123629 TI - Return of depressed men: changes in distribution of depression and symptom cases in Norway between 1990 and 2001. AB - BACKGROUND: Women's elevated risk of depression compared to men is a common finding in psychiatric epidemiology. Studies conducted in the 1950s and 1970s, however, documented approximately equal prevalence of sex rates. AIM: This study investigated changes in depression rates between 1990 and 2001 in Norway. Further, differences in severity were examined between men and women. METHOD: A study population (later to be called OsLof) was established in 1990, supplemented in 2001, and analysed as two cross-sectional datasets to compare sex and age differences in ICD-10 diagnoses and HSCL-25> or =1.75 caseness. Organic depression and the DSM-IV Major Depressive Episode were also examined in 2001. RESULTS: There was no significant change in the prevalence of depression or HSCL 25 caseness from 1990 to 2001. However, a major shift in prevalence occurred over time within the 18-34 year old age category with significantly higher rates among younger men (1% to 10%) and lowered rates among younger women (10% to 4%). Among persons depressed, no significant differences were found for severity between the sexes. LIMITATIONS: A selection of healthier participants than found in the source population might have resulted in lower prevalence than real. CONCLUSION: There was no indication of an overall increase in depression from 1990 to 2001, neither by diagnosis nor by symptom checklist scores. However, a major shift in prevalence occurred within the 18-34 year old age category with significantly higher rates among younger men and lowered rates among younger women. No sex differences existed in terms of severity of depression. PMID- 17123630 TI - Free T4 index and clinical outcome in patients with depression. AB - INTRODUCTION: Many studies of patients with major depression have reported that a significant decrease in serum free T(4) index is associated with specific treatments; what has been unanswered is whether these observations are generalizable. This study evaluated baseline thyroid function and its relationship to rapidity of treatment response as measured by hospital length of stay (HLOS). METHODS: Admission thyroid indices and HLOS data were harvested from the medical record of patients hospitalized for depression, both unipolar and bipolar. The relationship between admission thyroid indices and HLOS was evaluated using survival models. RESULTS: Controlling for age and year of discharge, an inverse relationship between FT(4) index (FT(4)I) and HLOS was present in men, but not in women. The mean HLOS stratified by gender and median FT(4)I was 50% shorter in men with a relatively high FT(4)I in comparison to the other three groups. LIMITATIONS: This is a retrospective study limited by the absence of a structured diagnostic psychiatric interview and prospective controlled antidepressant evaluation. CONCLUSION: These data would suggest that a relatively elevated FT(4) index in depressed men is associated with a faster antidepressant response time. Prospective study is encouraged to further clarify this potential thyroid/gender relationship and whether thyroid supplementation can accelerate the improvement in depression as measured by HLOS. PMID- 17123631 TI - The French version of the validated short TEMPS-A: the temperament evaluation of Memphis, Pisa, Paris and San Diego. AB - The TEMPS developed from classical temperament concepts at the Universities of Tennessee (Memphis) and California (San Diego) in collaboration with clinical scientists in Pisa and Paris. It presently exists in 20 languages and full validation of its 110-item version has been accomplished in American English, Italian, French, German, Hungarian, Japanese, Turkish, Lebanese Arabic and Argentinean Spanish. For many studies, a shorter version is easier to use. Accordingly, the 39-item validated English version has just been rendered into French, to facilitate clinical use and research in Francophone countries. PMID- 17123632 TI - Special issue on circular insanity and beyond: historic contributions of French psychiatry to contemporary concepts and research on bipolar disorder. AB - The original contribution of French psychiatry to what today we call bipolar disorder has its roots in 1854 in two papers that were presented in the French National Academy of Medicine by Falret and Baillarger. This event was celebrated in the same Academy in Paris in February, 2004 at its 150th anniversary. Because nearly all of the contributions of French alienists to the field of mood disorders have not been translated into English, their pioneering contributions are not as well known to the Anglophone world as those of Kraepelin, who in 1899 integrated French and other German work into his systematic treatise. To the best of our knowledge, the present special issue is the first monograph in English devoted to the pioneering French contributions in the field of mood disorders as well as contemporary research on bipolar disorder in France, Italy and the United States which supports and extends them. PMID- 17123633 TI - Clear distinction between preattentive and attentive process in schizophrenia by visual search performance. AB - Visual information-processing deficits were investigated in patients with schizophrenia using visual search tasks. Subjects comprised 20 patients with schizophrenia and 20 normal subjects. Visual search tasks were modified from those used previously to reveal more distinct differences between feature and conjunction search tasks. The presentation area of items in the present study was more than double the area used in our previous study [Mori, S., Tanaka, G., Ayaka, Y., Michitsuji, S., Niwa, H., Uemura, M., Ohta, Y., 1996. Preattentive and focal attentional processes in schizophrenia: a visual search study. Schizophrenia Research 22, 69-76], and items were distributed over the area randomly in each trial to produce a certain range of locational jitter for each item across trials that prevented a matrix-like presentation of items at fixed positions [Mori, S., Tanaka, G., Ayaka, Y., Michitsuji, S., Niwa, H., Uemura, M., Ohta, Y., 1996. Preattentive and focal attentional processes in schizophrenia: a visual search study. Schizophrenia Research 22, 69-76]. The target was a red square, and distractors were red circles in the feature search task and red circles and green squares in the conjunction search task. Slopes and intercepts of a linear function relating reaction times to set size were computed. In the feature search task, slopes for both groups were almost zero. In the conjunction search task, significant differences in slopes were seen between the two groups irrespective of target presence or absence. Moreover, the slopes were approximately twice as steep during target absence as during target presence. These results indicate more definitively than the results of our previous study [Mori, S., Tanaka, G., Ayaka, Y., Michitsuji, S., Niwa, H., Uemura, M., Ohta, Y., 1996. Preattentive and focal attentional processes in schizophrenia: a visual search study. Schizophrenia Research 22, 69-76] that patients with schizophrenia have deficits in focal attentional processing, although their preattentive processing functions at a normal level. PMID- 17123634 TI - Cognitive and emotion recognition deficits in obsessive-compulsive disorder. AB - Previous investigations have demonstrated impaired recognition of facial affect and cognitive dysfunction in several psychiatric disorders. The specificity of these deficits is still debated. The aim of this study was to investigate the deficits in emotion recognition and cognition in obsessive-compulsive disorder (OCD). Forty patients with OCD (DSM-IV, 16 women, 34.7+/-10.4 years) and 40 healthy volunteers (16 women, 34.7+/-8.7 years) were compared. All participants underwent a computerized neuropsychological test battery (Gur, R.C., Erwin, R.J., Gur, R.E., Zwil, A.S., Heimberg, C., Kraemer, H.C., 1992. Facial emotion discrimination II. Behavioral findings in depression. Psychiatry Research 42, 241 251; Gur, R.C., Ragland, J.D., Moberg, P.J., Turner, T.H., Bilker, W.B., Kohler, C., Siegel, S.J., Gur, R.E., 2001. Computerized neurocognitive scanning: I. Methodology and validation in healthy people. Neuropsychopharmacology 25, 766 776). A German version of the Penn Facial Emotion Acuity Test, the Facial Emotion Intensity Differentiation, including happy, sad and neutral faces, and the Facial Memory Test were administered. Executive functions were assessed by a computerized version of the Wisconsin Card Sorting Test and attention was evaluated using the Continuous Performance Test. OCD patients performed more poorly than healthy controls in facial memory tests (especially delayed), as well as in attention and executive functions. The only significant difference between the groups in emotion processing was poorer recognition of sad female faces in patients, who misperceived neutral faces as sad. The results point to memory and executive deficits in addition to a "negative" bias in emotion recognition in OCD patients. PMID- 17123635 TI - Hypercholesterolemia in Asperger syndrome: independence from lifestyle, obsessive compulsive behavior, and social anxiety. AB - We report on elevated total cholesterol and low-density lipoprotein (LDL) levels in 22 individuals with Asperger syndrome compared with well-matched controls, after accounting for lifestyle variables and clinical symptomatology that could affect them. A potential role for dyslipidemia in the pathogenesis of some forms of autism is discussed. PMID- 17123636 TI - Human T cell leukemia virus type I is resistant to the antiviral effects of APOBEC3. AB - The objective of this study was to investigate whether the anti-retroviral cellular cytidine deaminase, APOBEC3, inhibits the infectivity of human T cell leukemia virus type I (HTLV-I). Sufficient quantities of cell-free HTLV-I virion for infection were obtained by cotransfecting cells with HTLV-I and human or murine APOBEC3 expression vectors along with a plasmid expressing Tax. HTLV-I viruses containing these deaminases were still capable of infecting 293T and MOLT 4 cells. No G-to-A mutations, which are characteristic of cytidine deaminases, were observed in the HTLV-I genome. These results suggest that the enzymatic activity of APOBEC3 may not contribute substantially to antiviral responses to HTLV-I. PMID- 17123637 TI - A one-step, gel-based RT-PCR assay with comparable performance to real-time RT PCR for detection of classical swine fever virus. AB - Classical swine fever, a notifiable disease to the Office International des Epizooties (OIE), is a highly contagious viral disease affecting both domestic pigs and wild boars. Rapid, sensitive, and specific detection of the causing agent classical swine fever virus (CSFV) is therefore essential for diagnosis and control of the disease. Most protocols for gel-based PCR consist of two steps, reverse transcription followed by PCR. Such a protocol is time consuming, laborious and more prone to contamination. Two highly sensitive and fast one-step RT-PCR assays were developed for gel-based and real-time detection of CSFV, and their performances were compared to that of a published real-time assay. The results showed that the gel-based assay had comparable performance to the real time RT-PCR assays for detection of the virus. A detection limit of 50 copies was achieved by both assays. It is concluded that the one-step gel-based RT-PCR assay provides the simplest and most sensitive method for detection of CSFV in cell culture material or clinical samples, that can be applied in laboratories without facilities for real time PCR assays. The one-step format minimizes the risk for cross contamination and the hands-on time. The real-time assay is suitable for high-throughput screening of the virus in large populations. PMID- 17123638 TI - Oral 1-O-octadecyl-2-O-benzyl-sn-glycero-3-cidofovir targets the lung and is effective against a lethal respiratory challenge with ectromelia virus in mice. AB - Hexadecyloxypropyl-cidofovir (HDP-CDV) has been shown to be orally active against lethal infection with orthopoxviruses including, mousepox, cowpox, vaccinia and rabbitpox. The alkoxyalkyl group provides oral absorption and reduces greatly the amount of drug reaching the kidney, the site of CDV's dose limiting toxicity. However, the amount of HDP-CDV detected in lung, an important site of early poxvirus replication, is low and the reduction of viral titers in surviving animals is reduced moderately compared with the liver where poxvirus titers are virtually undetectable. We synthesized a novel glycerol ester of CDV, 1-O octadecyl-2-O-benzyl-sn-glycero-3-CDV (ODBG-CDV), and compared its oral pharmacokinetics with that of HDP-CDV. Surprisingly, ODBG-CDV levels in lung are much higher and liver levels are reduced, suggesting that the compound is transported in small intestinal lymph instead the portal vein. ODBG-CDV has excellent in vitro activity in cells infected with ectromelia virus (ECTV). In mice infected with a lethal aerosol or intranasal challenge of ECTV, HDP-CDV and ODBG-CDV are equally effective in preventing death from disease. Other drugs esterified to 1-O-octadecyl-2-O-benzyl-sn-glycerol or 1-O-octadecyl-2-O-benzyl-sn glycerol-3-phosphate may provide lung targeting for treatment of microbial or neoplastic diseases while reducing first pass removal by the liver during oral absorption. PMID- 17123639 TI - Activation in extended amygdala corresponds to altered hedonic processing during protracted morphine withdrawal. AB - Previously we reported that during protracted morphine abstinence rats show reduced conditioned place preferences (CPP) for food-associated environments, compared to non-dependent subjects. To determine the brain regions involved in this altered reward behavior, we examined neural activation (as indexed by Fos like proteins) induced by a preference test for a food-associated environment in 5-week morphine-abstinent versus non-dependent animals. The results indicate that elevated Fos expression in the anterior cingulate cortex (Cg) and basolateral amygdala (BLA) correlated positively with preference behavior in all groups. In contrast, Fos expression in stress-associated brain areas, including the ventral lateral bed nucleus of the stria terminalis (VL-BNST), central nucleus of the amygdala (CE), and noradrenergic (A2) neurons in the nucleus tractus solitarius (NTS) was significantly elevated only in morphine-abstinent animals. Furthermore, the number of Fos positive neurons in these areas was found to correlate negatively with food preference in abstinent animals. These results indicate that the altered hedonic processing during protracted morphine withdrawal leading to decreased preference for cues associated with natural rewards may involve heightened activity in stress-related brain areas of the extended amygdala and their medullary noradrenergic inputs. PMID- 17123640 TI - Adenosine A2A receptor blockade before striatal excitotoxic lesions prevents long term behavioural disturbances in the quinolinic rat model of Huntington's disease. AB - Huntington's disease (HD) is a progressive neurodegenerative disorder, characterised by severe degeneration of basal ganglia, motor abnormalities, impaired cognitive function and emotional disturbances. Many of the distinct neuropathological features of HD are reproduced in rats by intrastriatal injections of the excitotoxin quinolinic acid (QA), and QA-induced excitotoxicity is partially prevented by administration of the A(2A) receptor antagonist prior to the QA injection. In this study, we assessed the neuroprotective effects of the adenosine A(2A) receptor antagonist SCH 58261 on the progressive behavioural alterations reported in the QA rat model of Huntington's disease. Male rats received i.p. SCH 58261 (0.01mg/kg) or vehicle 20min before a bilateral injection of quinolinic acid (QA, 300nmol/1mul) or its vehicle in the dorsal striatum. Motor activity and anxiety levels were analyzed in an open-field arena and in an elevated plus-maze at 2 weeks, 2 months and 6 months post-lesion. In QA-lesioned rats SCH 58261 prevented alterations of wall rearing behaviour starting from 2 weeks post-lesion while emotional changes (reduced anxiety) were back to control levels by 6 months post-lesion. These findings extend to the behavioural parameters the protective effects of SCH 58261 in the QA model of Huntington's disease. PMID- 17123641 TI - Influence of gender on working and spatial memory in the novel object recognition task in the rat. AB - Gender differences in many behavioural tasks have been observed in both humans and laboratory animals. The novel object recognition (NOR) task is increasingly used to investigate drug effects on working memory processes, although, the influence of sexually dimorphic behaviours have not yet been evaluated. In addition, the role of natural fluctuations in the sex steroids during the oestrous cycle has received little attention during object recognition tasks. Therefore, the aim of the current study was to investigate the influence of gender and oestrous cycle phase on working and spatial memory using the NOR task. Animals were tested in the NOR task and the spatial NOR task. Male and female rats completed an acquisition trial followed by an inter-trial interval of a specified length, then a final retention trial. Vaginal cytology enabled the influence of oestrous cycle phase to be determined in both the NOR and spatial NOR, each animal was tested during one phase of their regular oestrous cycle only. It was found that female rats performed significantly better than male rats in the standard NOR paradigm (p<0.05 compared to no significance (NS) at 3h, respectively), while male rats showed improved memory in the spatial NOR paradigm compared with female rats (p<0.05 compared to NS at 3h, respectively). There was no influence of phase of oestrous cycle on the NOR task, however, during the spatial NOR there was a significant improvement in ability when oestrogen and progesterone levels have been shown to be at their lowest (i.e. p<0.05 during oestrous compared to NS at other stages). In conclusion, it is clear that gonadal hormones can influence components of memory and gender is an important consideration in experimental design. PMID- 17123642 TI - Thiols in Scenedesmus vacuolatus upon exposure to metals and metalloids. AB - Phytochelatins are intracellular metal ligands produced by algae when exposed to elevated metal concentrations. In freshwater ecosystems, algae are exposed to a wide range of metals and metalloids. The aim of this study was thus to investigate phytochelatin induction in freshwater algae upon metal and metalloid exposure. To that purpose, the unicellular green alga Scenedesmus vacuolatus, was exposed to Cu, Zn, Ni, Pb and Ag, as well as to As(III), As(V), Sb(III) and Sb(V), and examined for its thiol content (gamma-glutamylcysteine, glutathione and phytochelatins). Glutathione content was found to decrease upon the exposure to Zn and to increase upon the exposure to Pb and Ag. Phytochelatins were only induced by Cu (at [Cu2+] = 8x10(-11) M) and Pb (at [Pb2+] = 8x10(-11) to 8x10( 10) M), where [Cu2+] and [Pb2+] are computed free metal ion concentrations. Glutathione content also decreased upon the exposure to Sb(V) whereas an increase was observed as a result as the exposure to As(III) and As(V). The metalloids As(III), As(V) and Sb(III) in the concentration range from 8x10(-6) to 2x10(-4) M (total concentrations of oxyanions) were inducing phytochelatins. Glutathione and phytochelatin content in S. vacuolatus do thus sensitively respond to exposure to a number of metals and metalloids. PMID- 17123643 TI - Point mutations in mitochondrial topoisomerase enzymes of Trypanosoma congolense are not involved in isometamidium resistance. PMID- 17123644 TI - Oxidative stress impairs heme detoxification in the midgut of the cattle tick, Rhipicephalus (Boophilus) microplus. AB - In the cattle tick Rhipicephalus (Boophilus) microplus digestion of blood is intracellular, accomplished by the so-called digest cells that fill the midgut lumen. Hydrolysis of hemoglobin in the digestive vesicles of these cells results in the release of large amounts of heme, a pro-oxidant compound, whose iron atom, together with H(2)O(2), may participate in the Fenton reaction and lead to the production of hydroxyl radicals. Here, we investigated the role of catalase, an enzyme responsible for H(2)O(2) detoxification. Fully engorged female ticks injected with 3-amino-1,2,4-triazole (AT), a catalase inhibitor, showed increased H(2)O(2) in the gut, together with diminished life span and lower egg-laying rates. Increased mortality observed upon AT injection was reversed by further injection of exogenous catalase, 2 days after AT treatment, confirming that increased death was due to inhibition of this enzyme by AT. In primary cultures of digest cells, intracellular H(2)O(2) is limited to specific organelles, while treatment with AT in vitro resulted in increased H(2)O(2) spreading all over the cell, confirming the role of catalase in regulating H(2)O(2) levels. Ticks fed on a calf that had been injected with AT showed marked inhibition of catalase activity in the gut and diminished life span, oviposition and engorgement. Digest cells of these ticks had an altered morphology, showing heme spread all over the cytosol, instead of being limited to the hemosomes. The amount of aggregated heme found in isolated hemosome was also strongly decreased in AT-treated cattle. All together, our results indicate that catalase performs an important role in the control of redox balance in R. microplus, which dramatically affects hemosome formation and stability. This enzyme may be a target in the development of new methods for tick control. PMID- 17123645 TI - Raf kinase inhibitor protein affects activity of Plasmodium falciparum calcium dependent protein kinase 1. AB - Proteins, such as the raf kinase inhibitory protein (RKIP), serve as modulators of signalling pathways by either promoting or inhibiting the formation of productive signalling complexes through protein-protein interactions. In the present study, the plasmodial RKIP ortholog, PfPE-PB1, was cloned, recombinantly expressed and purified to homogeneity. The purified protein was used to investigate the effect of plasmodial RKIP on the autophosphorylation and substrate phosphorylation activity of Plasmodium falciparum calcium-dependent protein kinase 1, PfCDPK1. Phosphorylation of RKIP by PfCDPK1 in in vitro kinase assays suggests that RKIP may be an in vivo substrate of this kinase, although the specific activity of PfCDPK1 is approximately seven-fold lower when RKIP, instead of casein, an exogenous substrate of this enzyme, is used as a substrate. In addition to the observed phosphorylation of RKIP itself, its presence in the assays greatly enhanced the autophosphorylation capacity of PfCDPK1 by approximately 5.5-fold. This substantial increase in autophosphorylation activity was associated with a diminished substrate phosphorylation activity of PfCDPK1 when casein was used. At the same time, RKIP phosphorylation slightly increased when casein was included into the assays. Thus, RKIP is recognized as a substrate under in vitro conditions and appears to act as a regulator of PfCDPK1 kinase activity, which possibly is one of its actual functions in the parasite. PMID- 17123646 TI - Ontogeny of prolyl endopeptidase and pyroglutamyl peptidase I in rat tissues. AB - Prolyl endopeptidase and pyroglutamyl peptidase I are enzymes which participate in the degradation of thyrotropin-releasing hormone (TRH), a hormone which is thought to play an important role in the development of organs and tissues. Here, we have characterized the ontogeny of TRH degrading enzyme activity in the brain cortex, lung, heart, kidney and liver. Overall, prolyl endopeptidase activity was found to be 2 to 5 fold higher in newborn vs. adult rat tissues, with the exception of the soluble form in the liver and the particulate form in the lung. In contrast, the developmental profile of pyroglutamyl peptidase I activity was found to be more variable and tissue dependent. These results corroborate the idea that both enzymes play important, tissue-specific roles during the development and maturation of rat organs. PMID- 17123647 TI - Molecular cloning and characterization of human Aph2 gene, involved in AP-1 regulation by interaction with JAB1. AB - A human Aph2 gene (hAph2) was identified and cloned from a human placenta cDNA library. Bioinformatics analysis revealed hAPH2 protein shares 96% identity with mouse APH2 and contains a zf-DHHC domain (148-210aa), which is always involved in protein-protein or protein-DNA interaction. Differential expression patterns of hAph2 mRNA were observed in normal human tissues. Yeast two-hybrid screening found another hAPH2-interacting protein JAB1. The zf-DHHC domain of hAPH2 and the C-terminal of JAB1 were confirmed to be critical for the interaction. Fused with GFP and expressed in COS-7, NIH/3T3 and SMMC-7721 cell lines, hAPH2 showed predominant distribution in the cytoplasm and co-localized with JAB1 around the nucleus. Furthermore, overexpression of hAPH2 could increase apoptosis of COS-7 cells and negatively regulate JAB1-induced activation of AP-1 in a concentration dependent manner. The expression level of c-jun was also down-regulated by overexpression of hAPH2 in COS-7 cells. These data showed some basic characterization and function of hAph2 (hAPH2), dependent or independent with JAB1. PMID- 17123649 TI - Large recesses associated with left ventricular noncompaction: an unusual presentation of the disease. AB - Left ventricular noncompaction (LVNC) represents an arrest of myocardial morphogenesis, resulting in persistence of multiple prominent ventricular trabeculations and deep intertrabecular recesses. We report a case of a 30-year old patient who was referred to our hospital due to familial history of cardiomyopathy. The patient underwent transthoracic echocardiography, cardiac catheterization and contrast ventriculography and cardiac magnetic resonance imaging. All imaging techniques showed the characteristic pattern of noncompaction together with two unusually large recesses in the inferior wall of myocardium. This unusual form of LVNC may have an impact on the clinical course of the disease and may require anticoagulation therapy. PMID- 17123648 TI - Dusty protein kinases: primary structure, gene evolution, tissue specific expression and unique features of the catalytic domain. AB - Ser/Thr- and Tyr-Protein kinases constitute a key switch underlying the dynamic nature and graded regulation of signal transduction and pathway activities in cellular organization. Here we describe the identification and characterization of Dusty, a single-copy gene that arose in metazoan evolution and encodes a putative dual Ser/Thr and Tyr protein kinase with unique structural features. Dusty is widely expressed in vertebrates, broadly distributed in the central nervous system, and deregulated in certain human cancers. Confocal imaging of transiently expressed human Dusty-GFP fusion proteins showed a cytoplasmic distribution. Dusty proteins from lower to higher species display an increasing degree of sequence conservation from the N-terminal non-catalytic domain to C terminal catalytic domain. The non-catalytic region has eight conserved cysteine residues, multiple potential kinase-docking motifs and phosphorylation sites, whereas the catalytic domain is divergent and about equally distant of Ser/Thr and Tyr protein kinases. Homology analyses identified the essential catalytic residues, suggesting that Dusty homologues all possess the enzymatic activity of a protein kinase. Taken together, Dusty is a unique evolutionarily selected group of divergent protein kinases that may play important functional roles in the brain and other tissues of vertebrates. PMID- 17123650 TI - CT vs conventional angiography in unselected patients with suspected coronary heart disease. AB - BACKGROUND: To provide confirmatory evidence that the accuracy of MSCT coronary angiography is comparable to that of conventional coronary angiography. METHOD: 19 patients who had both MSCT and conventional coronary angiography were included in our study. The mean age of our patients was 62.3 (SD 7.7) and 42% of our patients were female. The coronary vascular tree was subdivided into 13 segments (modified from the 16-segment AHA classification). The grade of disease was classified into 1 of 6 categories; (1)0-50% stenosis; (2) 50-75% stenosis; (3) 75 99% stenosis; (4) Occluded; (5) aneurysmal; (6) fistula. The results of the imaging studies were analysed by two radiologists, who were blinded to the clinical data. RESULTS: A total of 172 coronary artery segments were visualised on both CT and conventional angiography. There is excellent statistical correlation between the appearance of the coronary arteries on MSCT and conventional angiography, as determined by Spearmans test (r=0.727, P<0.0001). MSCT coronary angiography had a sensitivity of 80.0%, specificity of 95.9%, NPV of 97.9% and PPV of 66.7%. CONCLUSION: The specificity and NPV from our study are similar to those obtained by previous researchers. This study confirms that a negative CT coronary angiogram is useful to rule out the presence of significant coronary heart disease. PMID- 17123651 TI - Maasai perception of the impact and incidence of malignant catarrhal fever (MCF) in southern Kenya. AB - We investigated the perceived impact of malignant catarrhal fever (MCF) to pastoralists in Isinya Division, a wildlife dispersal area of Nairobi National Park, and used a range of participatory epidemiology methodologies. We compared the relative importance, incidence and impact of MCF compared to other locally defined important diseases with a total of 158 respondents in 11 group meetings and 21 household meetings in July 2004. Direct losses due to disease were investigated through lowered prices as a result of the emergency sale of disease infected animals. Overall, Maasai in Isinya Division perceived east coast fever (ECF) to be the most important cattle disease and to have the highest incidence. Anthrax was considered to have the largest impact. In areas within or adjacent to the wildebeest calving zone, MCF was perceived to be the most important cattle disease and also to have the largest impact. Outside the calving zone, MCF was considered the fourth-most important disease with the fourth largest impact, and these were areas where wildebeest were less common. MCF was also the fourth-most common disease, and across the Division incidence was estimated at 5% in calves and 10% in adults. However, MCF incidence varied greatly throughout the study area, from 3% to 12%, and the highest incidence risks were found in areas where wildebeest came to calve. The percent drop in sale price per animal infected with MCF was estimated at 50% for MCF for the year 2003-2004. Forced avoidance movements away from wildebeest calves were reported to decrease livestock production due to loss of access to prime grazing sites. As suggested by pastoralists in this study, the development of compensation schemes or incentives from wildlife would reduce the conflict between livestock keeping and wildlife conservation. PMID- 17123652 TI - Exploring preference anomalies in double bounded contingent valuation. AB - Double bounded dichotomous choice (DBDC) contingent valuation offers increased efficiency of willingness to pay (WTP) estimates compared with the single bounded format. However, evidence suggests DBDC generates anomalous respondent behaviour. This paper provides the first investigation and explanation of these anomalies in health. Results suggest the incentives for truthful preference revelation are altered in the presence of a follow up question. This result is found using both regression techniques and analysis of raw responses. Although findings suggest 'very certain' respondents exhibit less anomalous behaviour inconsistencies remain across bounds. The results of this study question the use of iterative valuation formats. PMID- 17123653 TI - Molecular differentiation of common promoters in Salmonella class 1 integrons. AB - The integron is a mobile gene element which harbors antibiotic-resistance gene cassettes capable of site-specific integration. Among the four known types of integrons, the class 1 integron has been associated with multidrug-resistance in pathogenic bacteria. These gene cassettes have been the focus of a series of studies. The gene cassettes share a common promoter, and their expression levels are affected not only by their proximity to the promoter, but also by the strength (weak, hybrid and strong) of the common promoter, P1, as well as the presence of the additional promoter, P2. In this study, we developed molecular methods for the differentiation of promoter structures using PCR, restriction enzyme analysis, and polyacrylamide gel electrophoresis, and have applied them to the characterization of class 1 integrons in 33 non-typhoidal Salmonella serotypes in Korea. Class 1 integrons were detected in four serotypes: S. Derby (SD), S. Istanbul (SI), S. Paratyphi B (SPB), and S. Livingstone (SL), and the amplicon sizes were 1.0 Kb (SD, SI and SPB) and 2.0 Kb (SL). All of the 1.0 kb amplicons harbored gene cassettes (aadA1 or aadA2), but the 2.0 kb amplicon harbored three (dhfrXII-orfF-aadA2) gene cassettes, which conferred streptomycin/spectinomycin (aadA) and trimethoprim (dhfr) resistances. Our promoter structure study revealed three types of promoters; strong P1 (SD), weak P1 (SPB and SL), and weak P1+P2 (SI). In conclusion, the class 1 integrons were detected in Korean NTS, and their promoter structures were found to be variable. Therefore, our methods may prove helpful in terms of our understanding of molecular diversity, as well as the transmission of class 1 integrons and phenotype-genotype relationships in antibiotic-resistance. PMID- 17123654 TI - Lack of efficacy of music to improve sleep: a polysomnographic and quantitative EEG analysis. AB - An increasing number of studies have been examining non-pharmacological methods to improve the quality of sleep, including the use of music and other types of auditory stimulation. While many of these studies have found significant results, they suffer from a combination of subjective self-report measures as the primary outcome, a lack of proper controls, often combine music with some type of relaxation therapy, or do not randomise subjects to control and treatment conditions. It is therefore difficult to assess the efficacy of music to induce or improve sleep. The present study therefore examined the effects of music using standard polysomnographic measures and quantitative analysis of the electroencephalogram, along with subjective ratings of sleep quality. In addition, a tones condition was used to compare any effects of music with the effects of general auditory stimulation. Using a counter-balanced within-subjects design, the music was not significantly better than the tones or control conditions in improving sleep onset latency, sleep efficiency, wake time after sleep onset, or percent slow wave sleep, as determined by objective physiological criteria. PMID- 17123655 TI - Event-related potentials during an emotional Stroop task. AB - Emotional Stroop tasks have gained wide interest in scientific literature in the last two decades. Although no direct measure of attention is employed, these studies infer the presence of preferential processing of threatening information based on reaction time (RT) impairment in a competing task. Because event-related potential (ERP) measures are sensitive to both the extent (amplitude) and speed (latency) of cerebral processing, they are valuable tools with which to examine more directly the claim that threatening stimuli are associated with enhanced attention. Twenty-two students rated a pool of words to identify those that were personally disturbing. Two word types (threat and neutral) were then compared in two tasks (color relevant, in which the color ink of words was identified, and word relevant in which words were classified as threatening or not). No emotional Stroop effect was observed in terms of longer RTs to identify the colors of threat words. ERP results provided valuable information about threat processing which was not observed with behavioral measures. Threat content was associated with larger P2 amplitude in the right than left hemisphere, and larger P3 amplitude, across tasks. The results indicate strong evidence for enhanced processing of threat-related stimuli in healthy individuals. It is concluded that ERPs are a sensitive measure of processes underlying emotional Stroop performance, which can be used to elucidate attentional biases in healthy and clinical populations. PMID- 17123656 TI - Sequence polymorphisms in the long terminal repeat of bovine leukemia virus: evidence for selection pressures in regulatory sequences. AB - Bovine leukemia virus (BLV) is an oncogenic virus widespread in cattle. It belongs to the genus Deltaretrovirus of the family Retroviridae along with human and simian T-lymphotropic viruses. The BLV transcriptional promoter is located in the proviral 5' long terminal repeat (LTR), composed of U3, R, and U5 regions. BLV LTR contains multiple cis-acting elements important for promoter activity, a short coding sequence (encoding the NH(2) terminus of the G4 regulatory protein), and non-regulatory/non-coding regions. Variation in coding sequences of BLV structural proteins has been studied extensively, but little work has been done on sequence variability of non-coding regions, mostly located in LTR. Here, we report the first study on the natural diversity of the BLV LTR, using viral isolates from 52 cattle in several different areas worldwide. Nucleotide variations from the consensus sequence were observed in most isolates and clustered phylogenetically, corresponding to the geographic distribution of donor cattle. Overall, regulatory regions were significantly more conserved than non regulatory regions in the BLV LTR, as well as in LTR sub-regions (U3, R, and U5). Evidence of selection pressures in BLV LTR suggests that selection occurs not only in coding sequences, but may also involve regulatory sequences. PMID- 17123657 TI - Generation of henipavirus nucleocapsid proteins in yeast Saccharomyces cerevisiae. AB - Hendra and Nipah viruses are newly emerged, zoonotic viruses and their genomes have nucleotide and predicted amino acid homologies placing them in the family Paramyxoviridae. Currently these viruses are classified in the new genus Henipavirus, within the subfamily Paramyxovirinae, family Paramyxoviridae. The genes encoding HeV and NiV nucleocapsid proteins were cloned into the yeast Saccharomyces cerevisiae expression vector pFGG3 under control of GAL7 promoter. A high level of expression of these proteins (18-20 mg l(-1) of yeast culture) was obtained. Mass spectrometric analysis confirmed the primary structure of both proteins with 92% sequence coverage obtained using MS/MS analysis. Electron microscopy demonstrated the assembly of typical herring-bone structures of purified recombinant nucleocapsid proteins, characteristic for other paramyxoviruses. The nucleocapsid proteins revealed stability in yeast and can be easily purified by cesium chloride gradient ultracentrifugation. HeV nucleocapsid protein was detected by sera derived from fruit bats, humans, horses infected with HeV, and NiV nucleocapsid protein was immunodetected with sera from, fruit bats, humans and pigs. The development of an efficient and cost-effective system for generation of henipavirus nucleocapsid proteins might help to improve reagents for diagnosis of viruses. PMID- 17123660 TI - Interactions between GSTM1, GSTT1 and GSTP1 polymorphisms and smoking and intake of fruit and vegetables in relation to lung cancer. AB - Polymorphisms in glutathione S-transferases (GST) are weakly associated with risk for lung cancer. We examined gene-environment interactions in relation to lung cancer in 430 cases and 767 comparison persons identified within a prospective cohort of 57,053 persons. We used assays capable of discriminating heterozygous individuals from those with two functional alleles and homozygous deletions in GSTM1 and GSTT1. There was no overall association between the GST polymorphisms and lung cancer. We found that fruit and vegetables reduced the risk of lung cancer only among carriers of at least one functional GSTM1 allele, and among carriers of two GSTP1 Val alleles. There were no significant interactions between the GST polymorphisms and smoking. PMID- 17123661 TI - Prognostic assessment after surgical resection for non-small cell lung cancer: experiences in 2083 patients. AB - The importance of the TNM staging system for patient management, clinical research and communicating information about lung cancer is of international importance. Modifications of the TNM classification system is scheduled for the near future. A retrospective review of 2376 patients with primary non-small cell lung cancer treated in a monocentric institution between 1996 and 2005 was performed. The overall 5-year survival rate was 46.8%. A total of 2083 patients had complete resections with a 5-year survival of 50.7%. After complete resection the 5-year survival rates by pathological stage of the disease were as follows: 68.5% for IA, 66.6% for IB, 55.3% for IIA, 49.0% for IIB, 35.8% for IIIA, 35.4% for IIIB, and not defined (3-year survival: 33.1%) for IV. The difference in prognosis between stage IIB and IIIA was significant (p=0.001) there was no significant difference between IA and IB, between IB and IIA, between IIA and IIB, between IIIA and IIIB, or between IIIB and IV. In stage IV there was a significant difference in survival between patients with pulmonary metastases or distant extrapulmonary metastases (p=0.001). In multivariate analysis, we also found gender and histology to be independent significant prognostic factors for survival. Multiple factors influence the long-term survival of patients with non small cell lung cancer after surgical resection. The present stage related prognosis seems to characterize patient prognosis and outcome reliable. For further data review there should be a focus on stage IV disease. PMID- 17123662 TI - Comment on "Analysis of groundwater contamination using concentration-time series recorded during an integral pumping test: Bias introduced by strong concentration gradients within the plume" by Allelign Zeru and Gerhard Schafer. AB - We consider the results of a recent paper in this journal [Zeru, A. and Schafer, G., 2005. Analysis of groundwater contamination using concentration-time series recorded during an integral pumping test: Bias introduced by strong concentration gradients within the plume. Journal of Contaminant Hydrology 81 (2005) 106-124], which addresses the field-scale characterisation of contaminant plumes in groundwater. There, it is concluded that contaminant concentration gradients can bias Integral Pumping Test (IPT) interpretations considerably, in particular if IPTs are conducted in advective fronts of contaminant plumes. We discuss implications of this setting and also argue that the longitudinal and transverse dispersivities used in the examples of Zeru and Schafer (2005) of up to 30 m and 3 m, respectively, are generally very high for the here relevant capture zone scale (<20 m). However, regardless of both longitudinal and transverse concentration gradients, we further show through a counter-example that IPT results are unbiased as long as the concentration attenuation along the flow direction is linear over the capture zone extent. PMID- 17123663 TI - Evidence against memorial facilitation and context-dependent memory effects through the chewing of gum. AB - The experiment examined the prediction that chewing gum at learning and/or recall facilitated subsequent word recall. Chewing gum at learning significantly impaired recall, indicating that the chewing of gum has a detrimental impact upon initial word encoding. In addition, a context-dependent memory effect was reported for those participants who both learned and recalled in the absence of gum; however, a context-dependent effect was not found with chewing gum. The findings contradict previous research. PMID- 17123658 TI - Display technologies: application for the discovery of drug and gene delivery agents. AB - Recognition of molecular diversity of cell surface proteomes in disease is essential for the development of targeted therapies. Progress in targeted therapeutics requires establishing effective approaches for high-throughput identification of agents specific for clinically relevant cell surface markers. Over the past decade, a number of platform strategies have been developed to screen polypeptide libraries for ligands targeting receptors selectively expressed in the context of various cell surface proteomes. Streamlined procedures for identification of ligand-receptor pairs that could serve as targets in disease diagnosis, profiling, imaging and therapy have relied on the display technologies, in which polypeptides with desired binding profiles can be serially selected, in a process called biopanning, based on their physical linkage with the encoding nucleic acid. These technologies include virus/phage display, cell display, ribosomal display, mRNA display and covalent DNA display (CDT), with phage display being by far the most utilized. The scope of this review is the recent advancements in the display technologies with a particular emphasis on molecular mapping of cell surface proteomes with peptide phage display. Prospective applications of targeted compounds derived from display libraries in the discovery of targeted drugs and gene therapy vectors are discussed. PMID- 17123664 TI - An antifungal peptide from red lentil seeds. AB - An antifungal peptide, with a molecular mass of 11 kDa, was isolated from dry seeds of the red lentil (Lens culinaris) using a procedure that involved four chromatographic steps. The antifungal peptide was unadsorbed on DEAE-cellulose, and adsorbed on Affi-gel blue gel and S-Sepharose. The final chromatographic step involved gel filtration by fast protein liquid chromatography on Superdex 75. The antifungal peptide inhibited mycelial growth in Mycosphaerella arachidicola with an IC50 of 36 microM. It also exhibited antifungal activity against Fusarium oxysporum, but there was no inhibitory activity toward tumor cell lines and human immunodeficiency virus type 1 reverse transcriptase (RT). PMID- 17123665 TI - Enhanced soluble CD40 ligand and Alzheimer's disease: evidence of a possible pathogenetic role. AB - It has been suggested that cerebrovascular factors contribute to Alzheimer's disease. Soluble CD40 ligand (sCD40L) is directly involved in the development of vascular damage. We tested the hypothesis that sCD40L may be enhanced in Alzheimer's disease and predictive of its clinical course. Plasma sCD40L levels were evaluated in three groups of 40 consecutive patients each referring for mild or moderate or severe Alzheimer's disease, as assessed by the Clinical Dementia Rating (CDR), and in 40 healthy subjects. Seventy-seven patients with mild or moderate disease were re-evaluated after 2 years. Cross-sectional comparisons revealed higher plasma sCD40L levels in Alzheimer's disease patients than in controls (9.3+/-4.7 ng/mL versus 3.4+/-1.3 ng/mL, p<0.0001). Circulating sCD40L levels significantly increased through the three CDR stages (p=0.0011 or less) and were correlated with MMSE (r=-0.574, p<0.0001) and ADAS-cog subscale (r=0.538, p<0.0001) scores. Longitudinal evaluation identified sCD40L as an independent predictor of MMSE (beta=-0.157, t=-3.650, p=0.0005) and ADAS-cog subscale (beta=0.484, t=3.890, p=0.0002) score changes after 2 years. Patients with plasma sCD40L level>or=6.0 ng/mL, identified by ROC curve analysis as the best discriminating value for disease progression, had a three-fold increase in the risk of progression toward a worse CDR stage (odd ratio: 3.0, C.I. 95% 1.2 8.1). In conclusion, circulating sCD40L is enhanced in patients with Alzheimer's disease and independently associated with the severity and progression of the disease. These data might suggest a pathogenetic role for sCD40L in Alzheimer's disease. PMID- 17123666 TI - Loss of responsiveness to melatonin in the aging mouse suprachiasmatic nucleus. AB - Melatonin modulates circadian rhythms via the hypothalamic suprachiasmatic nucleus (SCN). One of the most robust assays for SCN melatonin receptor activation in mice is the inhibition of PACAP-induced phosphorylation of the transcription factor Ca(2+)/cAMP responsive element binding protein (CREB). To assess the effect of aging on responsiveness to melatonin, SCN slices from mice of different ages were prepared and treated with PACAP alone or PACAP plus melatonin. CREB phosphorylation state was assessed by immunohistochemistry. In SCN slices from young (2-4-month-old) mice, melatonin reduced the level of phospho-CREB immunoreactivity following PACAP treatment in a dose-dependent manner. In SCN slices from aged mice (19-22 months of age), PACAP alone induced comparable levels of phospho-CREB, but melatonin treatment failed to inhibit the PACAP-induced CREB phosphorylation. The results indicate an age-related loss of sensitivity to melatonin in the SCN. The findings are discussed in the context of the impact of endogenous and exogenous melatonin on sleep in elderly humans. PMID- 17123667 TI - Perception of legal liability by registered nurses in Korea. AB - Liability to the nursing profession is imperative if nurses are to act as an autonomous body. Assessing and communicating effectively is a vital part of nursing for patient safety. This study was designed to identify the attitudes of Korean nurses toward liability in assessment and communication and to investigate the relationship among the variables (i.e., legal awareness, attitudes toward doctor's duty to supervise nurses). The attitudes toward doctor's duty reflect the status of nurses' dependency on doctor's supervision. The study participants were 288 registered nurses in RN-BSN courses at two colleges in Korea. The level of legal awareness was measured using a 25-item Legal Awareness Questionnaire developed by the authors. The measuring instrument for attitudes toward doctor's duty to supervise nurses and nurses' liability was the Attitude toward Duty and Liability Questionnaire, which was modified by the authors. There were significant correlation between attitude toward doctor's duty and nurses' liability, but not between legal awareness and liability attitude. The results of this study suggest that the present educational content aimed at improving liability attitudes of nurses should be refocused with attitude-oriented education and should include an understanding of the increased accountability that comes with greater autonomy in nursing practice. PMID- 17123668 TI - Evaluating nurse prescribers' education and continuing professional development for independent prescribing practice: findings from a national survey in England. AB - BACKGROUND: The number of nurses able to independently prescribe medicines in England has risen steadily in recent years. AIM: To evaluate the adequacy of nurses' educational preparation for independent prescribing and to describe nurses' experiences of their continuing professional development as prescribers in practice. DESIGN AND METHOD: Postal questionnaire survey. PARTICIPANTS: Random sample of 246 nurses registered as nurse independent prescribers with the Nursing and Midwifery Council. RESULTS: The majority of nurses considered that the initial taught course element of their education programme met their needs, either to some extent (61% 151/246), or completely (22% 54/246). Most nurses (77% 190/246) received the specified 12 days support from their supervising medical practitioner and most were satisfied and positive about this experience. Nearly all of the nurses (>95%) reported that they were able to maintain a range of specified prescribing competencies in practice. Two thirds (62% 152/246) of the sample reported that they were receiving support/supervision for prescribing. Ninety five per cent (233/246) of the sample also reported that they engaged in self-directed informal continuing professional development, but only half of the sample had experience of formally provided professional development opportunities. Approximately half (52% 127/246) of the sample identified needs for continuing professional development. CONCLUSION: This first national survey of the education and professional development experiences of nurse independent prescribers in England provides evidence which highlights areas in which national policy is working well, and also points up issues which may need addressing as the roll out of nurse prescribing continues. The study also highlights characteristics and issues that health care policy makers and nurse educationalists internationally may wish to consider in developing and refining their own nurse prescriber education programmes. PMID- 17123669 TI - The knowledge, attitude and practice of the primary and secondary care nurse midwife practitioners on developmental dysplasia of hip. AB - A detailed clinical examination of the newborn performed by nurse-midwife practitioners has been recommended as a standard practice throughout the most western countries. The purpose of this study was to evaluate and compare the knowledge, attitude and practice of the primary and secondary care nurse-midwife practitioners on developmental dysplasia of hip (DDH) before and after a structured lesson. The study was designed as a cross-sectional, pre- and post test. The number of participants from primary care and secondary care nurse midwife practitioners were 155 and 208, respectively. A structured questionnaire consisting of 28 statements of medical and practical knowledge, and traditional attitudes with regard to DDH was administered. The results determined that the pre-test scores were much lower than the post-test scores of both primary and secondary care practitioners. Although there was a lack of knowledge about DDH in both groups, the pre-test and post-test scores were higher in the primary care group compared to the secondary care group. In conclusion, the knowledge, attitude and practice of the primary and secondary care nurse-midwife practitioners on developmental dysplasia of hip are in need of improvement. This improvement may be provided by continuing education programmes. PMID- 17123670 TI - Multiple CD4 and CD8 T-cell activation parameters predict vaccine efficacy in vivo mediated by individual DC-activating agonists. AB - A systematic comparison of the immunostimulatory capacity of TLR 2, 3, 4, 5, 7 and 9 agonists and an agonistic CD40-specific antibody was performed in a single long peptide vaccination model. All adjuvants activated DC in vitro but not all induced a strong functional T-cell response in vivo. Optimal clonal CD8(+) T-cell expansion depended on the capacity of agonists to mature pro-inflammatory DC and the duration of their in vivo stimulatory effect. Strong agonists promoted the induction of both antigen-specific IFNgamma-producing CD4(+) T-helper cells and high numbers of IFNgamma producing CD8(+) effector T-cells that killed target cells in vivo. Importantly, the capacity of an agonist to function as an adjuvant depended on the vaccine strategy used. Collectively, the multi-parameter system presented here can be used as a general road map to develop therapeutic vaccines. PMID- 17123671 TI - Cementifying fibroma of the frontal bone: a case report. AB - Cementifying fibromas are rare fibro-osseous tumours that are usually found in the jaws. We report a 10-year-old boy who presented with a mass in the frontal bone causing frontal bossing and proptosis. The preoperative diagnosis was not clear despite computed tomographic and magnetic resonance imaging. The lesion was resected completely and found to be a cementifying fibroma. It was reconstructed at the same operation and he made a good recovery with no sign of recurrence. PMID- 17123672 TI - Personality disorder, psychiatric symptoms and experience of childbirth among childbearing women in Sweden. AB - OBJECTIVE: to investigate the importance of having a personality disorder or psychiatric symptoms during pregnancy for women's experience of childbirth. DESIGN: a prospective study of women consecutively recruited during mid pregnancy. On study entry, the women completed self-rating scales for personality disorders, global functioning level and psychiatric symptoms. Two weeks after the birth, the women completed a questionnaire about their childbirth experiences. Obstetric and neonatal data were collected from maternity ward records. Multivariate logistic regression was used to estimate the likelihood for negative birth experiences. SETTING: antenatal clinics in Stockholm, Sweden. PARTICIPANTS: 625 primiparous women. FINDINGS: 40 out of 624 (6.4%) women fulfilled the criteria for personality disorder, and 28 out of 625 (4.5%) women were defined as psychiatric cases. No significant differences were found between women with personality disorders, or defined as psychiatric cases, and women without any mental problems regarding their global experience of birth. Women defined as psychiatric cases were more anxious (z=-2.5, p=0.04) during labour than women without mental problems. The strongest predictors of having had a negative global experience of birth were instrumental vaginal delivery (OR 3.2, 95% CI 2.1-5.1) and epidural analgesia (OR 2.4, 95% CI 1.5-3.9). KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: obstetric factors, such as instrumental delivery, seem to be more important than mental problems during pregnancy for women's global experience of birth. Women with psychiatric symptoms during pregnancy need extra support during labour. PMID- 17123673 TI - Comparison of forearm and conventional Bier's blocks for manipulation and reduction of distal radius fractures. AB - Forearm-based Bier's block has been advocated as a useful anaesthesic technique in hand surgery. However, there is limited data comparing forearm blocks with the conventional Bier's block. We conducted a randomised controlled trial (n=30) comparing the two techniques of anaesthesia for manipulation and reduction of closed distal radius fractures in an emergency room setting. Pain scores measured using the Visual Analogue Scale during the procedure were used as the primary outcome assessment. There was no significant difference in pain scores between the forearm and conventional Bier's block (mean VAS 18.4 SD 22.10 versus 33.7 SD 29.6). No major complications were observed in either group. The forearm-based Bier block is an effective alternative to the conventional block. PMID- 17123674 TI - Responsiveness of the Dutch version of the DASH as an outcome measure for carpal tunnel syndrome. AB - A cohort of 119 patients with carpal tunnel syndrome completed the questionnaire of the Dutch version of the DASH score pre-operatively and one year postoperatively. The mean DASH score decreased from 38.2 to 22.0. There was a significant correlation with the Boston carpal tunnel outcome score (r=0.78). With an effect size of 0.87 and a standardized mean response of 0.69, the Dutch version of the DASH is highly responsive for the evaluation of the outcome of surgery for carpal tunnel syndrome. PMID- 17123675 TI - Isolated tuberculosis of a metacarpal bone in a 2 year-old child. PMID- 17123676 TI - Neural correlates of inner speech and auditory verbal hallucinations: a critical review and theoretical integration. AB - The neuroimaging and neurophysiological literature on inner speech in healthy participants and those who experience auditory verbal hallucinations (AVHs) is reviewed. AVH-hearers in remission and controls do not differ neurologically on tasks involving low levels of verbal self-monitoring (VSM), such as reciting sentences in inner speech. In contrast, on tasks involving high levels of VSM, such as auditory verbal imagery, AVH-hearers in remission show less activation in areas including the middle and superior temporal gyri. This pattern of findings leads to a conundrum, given that mentation involving low levels of VSM is typically held to form the raw material for AVHs. We address this by noting that existing neuroimaging and neurophysiological studies have been based on unexamined assumptions about the form and developmental significance of inner speech. We set out a Vygotskian approach to AVHs which can account for why they are generally experienced as the voice of another person, with specific acoustic properties, and a tendency to take the form of commands. On this approach, which we argue is consistent with the neural correlates evidence, AVHs result from abnormalities in the transition between condensed and expanded dialogic inner speech. Further potential empirical tests of this model are discussed. PMID- 17123677 TI - Social support at work and the risk of myocardial infarction and stroke in women and men. AB - It has been proposed that lack of social support in a work place characterized by high levels of stress, may increase the likelihood of future cardiovascular disease. The aim of this study was to analyze the prospective impact of social support at work in combination with self-reported work stress on incidence of myocardial infarction (MI) and stroke in a cohort of 4707 women (mean age: 54.2 years) and 3063 men (mean age: 55.5 years) in Malmo, Sweden. The results are based on self-reports of work-related stress and social support collected at baseline examinations between the years 1992 and 1996. Work-stress was operationalized according to the Karasek job strain model. Data on incidence of MI and stroke were obtained from national and regional registers. At the end of follow-up, December 31, 2001, 38 women had experienced an MI and 53 had had a stroke. Corresponding figures for men were 114 MIs and 81 strokes. The first finding was that social support at work was an independent predictor of an MI and stroke among women. The second finding was that there was no evidence to support the iso-strain model. The third finding was that low levels of social support at work together with a passive work situation indicated an increased risk of a future cardiovascular outcome (MI or stroke) during follow-up in the female group. In men, no association was found between any psychosocial work conditions and incidence of MI or stroke during the same follow-up period. PMID- 17123678 TI - Sex in Geneva, sex in Lilongwe, and sex in Balaka. AB - This paper compares three interpretations of women and the transmission of HIV in rural Malawi. One is disseminated world-wide by institutions with a global reach such as the World Health Organization in Geneva and United States Agency for International Development (USAID); the second is provided by urban Malawians situated in the capital of Lilongwe, the seat of government and the site of the many international and national non-governmental agencies; the third is articulated by rural women and men in Balaka District, Malawi. We focus on women's motivations for extramarital sex, using qualitative interview data. We find that whereas in Geneva and Lilongwe women from rural Malawi are said to engage in sexual relationships outside of marriage because they need money for survival, in Balaka they are said to be motivated not only by money for survival but also for attractive consumer goods as well as by passion and by revenge for a husband's infidelity. We also find that data collection procedures influence explanations for women's affairs, as does the respondent's gender. An implication of this study is that AIDS-prevention policies based on the view from Geneva and Lilongwe need modification for Balaka, and by implication for rural Africa more generally. PMID- 17123679 TI - Meaning and coherence of self and health: an approach based on narratives of life events. AB - Using contributions from the fields of interactionist sociology and narrative studies and the results of an empirical study, in this article we examine the subject's mediation between life events and state of health. Examining narratives focusing on significant life events from 26 men and women aged between 51 and 73 from the administrative region of Ile de France, we suggest that dimensions of meaning and coherence of self need not be taken as internal dispositions, but rather as the product of sociocognitive work on self-narration. The characterisation of the types of causal relationships that the narrators establish between significant events and other events, situations or periods in their lives, opens up avenues for the development of a methodological tool to replace traditional ways of measuring sense of coherence and meaning/purpose in life using psychometric methods. This alternative perspective based on a conception of the self as narrative might provide a remedy for certain conceptual and methodological difficulties that are found within the field of coping research. PMID- 17123680 TI - Cohabitation, marriage, and 'sexual monogamy' in Nairobi's slums. AB - The current study investigates the extent to which sexual exclusivity--the restriction of one's sexual engagements to a single partner--prevails across various marital status, union type, and co-residence categories among Nairobi's poorest residents, slum dwellers. This question is central to the spread of HIV in the increasingly urban and poor, high prevalence countries of sub-Saharan Africa, where transmission is primarily via heterosexual sex. In many circles, sexual exclusivity is considered a prominent feature of the marriage institution. Yet, marriage and cohabitation are often not easily distinguishable in sub Saharan Africa, meaning that the frequent use, as a proxy, of the "in union" category, which includes married as well as cohabiting persons can, at best, be considered tenuous. Using the 2000 Nairobi Cross-Sectional Slum Survey (NCSS), this paper confirms that marriage is associated with higher reports of sexual exclusivity even in settings where poverty provokes risky behavior. The finding, here, is of lower risk of HIV infection for married respondents, with a smaller effect observed among non-married cohabiters. Converse to the implied benefits of marriage, though, women with co-wives are more likely to report multiple partners. The implications of these findings are discussed. PMID- 17123681 TI - Reboxetine induced erectile dysfunction and spontaneous ejaculation during defecation and micturition. AB - This is a case report of reboxetine induced erectile dysfunction, seminal emission and ejaculation during defecation and micturition. A 44 year old male who had been suffering from depression without any sexual dysfunction was put on venlafaxine XR treatment. Due to delayed ejaculation and occasional episodes of absence of ejaculation he was switched to reboxetine. At the second week of treatment he reported erectile dysfunction and premature ejaculation, and seminal emission and ejaculation during defecation and micturition occurred later at 8th week of treatment. After he was switched to sertraline 50 mg/day, his erectile dysfunction, premature and spontaneous ejaculation symptoms subsided in 2 weeks. Although reboxetine is reported to be free of sexual side effects, individual vulnerabilities to such unwanted effects should be considered, and sexual dysfunction should be assessed thoroughly during the treatment. PMID- 17123682 TI - High-dose ziprasidone-induced acute dystonia. AB - There are few clinical data describing the relative risks of extrapyramidal symptoms (EPS) at higher doses of ziprasidone (i.e., greater than 160 mg/day) when compared to lower doses. We report on a patient who displayed no EPS with ziprasidone 160 mg/day for several months, but did display marked, acute dystonic reactions on ziprasidone 240 mg/day. The observations from the present case indicate that high-dose, ziprasidone-induced acute dystonia may occur even if no such side effects have been experienced at doses up to and including 160 mg/day. PMID- 17123683 TI - In vitro model for decontamination of human skin: formaldehyde. AB - Decontamination of a chemical from skin is often an emergency measure. This study utilized an in vitro model to compare the decontamination capacity of three model decontaminant solutions (tap water, isotonic saline, and hypertonic saline). Human cadaver skin was dosed (approximately 0.25 microg on 3 cm(2) per skin) with radio-labeled [(14)C]-formaldehyde. After a defined exposure time (1, 3, and 30 min post-dosing, respectively), the surface skin was washed three times (4ml per time) with each solution. After washing, the skin was stripped with tape discs twice. Lastly, the wash solutions, strippings, receptor fluid, and remainder of skin were liquid scintillation analyzer counted to determine the amounts of formaldehyde. Additionally, an evaporation test at different exposure times (1min, 3min, 15min, 30min, and 60min, respectively) was conducted to monitor formaldehyde % evaporation. There were no statistical differences among these groups except isotonic saline, at 3min post-exposure (in wash solutions), showed a significantly difference (p<0.05) when compared to tap water. Formaldehyde % evaporation increased linearly with extending application times, and were 7.7%, 13.6%, 19.7%, 24.4%, and 35.9% (1min, 3min, 15min, 30min, and 60min, respectively). This data suggests that isotonic saline may be effective in removing formaldehyde from skin. However, results from this model need validation in vivo. The model may provide a facile and robust method of accelerating knowledge of decontamination mechanism and lead to enhanced efficacy. PMID- 17123684 TI - Effect of diallyl trisulfide-rich garlic oil on blood coagulation and plasma activity of anticoagulation factors in rats. AB - Diallyl trisulfide (DAT)-rich garlic oil was fed to Sprague-Dawley rats and the effects of this DAT-rich garlic oil on bleeding time, clotting time and anticoagulation factors were examined. Garlic oil supplement at 5 or 50mg garlic oil/kg bodyweight significantly prolonged bleeding time and thrombin time, and enhanced anticoagulation factor activity, such as antithrombin III and protein C (P<0.05). These results suggested that the anticoagulant action of DAT-rich garlic oil was due to inhibition and/or inactivation of thrombin. In addition, DAT-rich garlic oil benefits blood anticoagulation factors, which might further prevent the development of thrombus formation. However, the intake of garlic oil at high dose significantly increased plasma fibrinogen concentration (P<0.05), and affected the levels of several hematological parameters such as erythrocyte count, hemoglobin and platelets (P<0.05). The adverse effect of high doses of garlic oil might further influence the hemostatic balance. Therefore, the concentration of DAT-rich garlic oil should be carefully considered in its application. Supplementation of garlic oil at 5mg/kg bodyweight has anticoagulation effect in this animal study. PMID- 17123685 TI - [Treatment of naso-eyelids wrinkle mark through exeresis suture: preliminary series of nine cases]. PMID- 17123686 TI - Oxidative and nitrosative stress in trichloroethene-mediated autoimmune response. AB - Reactive oxygen and nitrogen species (RONS) are implicated in the pathogenesis of several autoimmune diseases. Also, increased lipid peroxidation and protein nitration are reported in systemic autoimmune diseases. Lipid peroxidation derived aldehydes (LPDAs) such as malondialdehyde (MDA) and 4-hydroxynonenal (HNE) are highly reactive and bind proteins covalently, but their potential to elicit an autoimmune response and contribution to disease pathogenesis remain unclear. Similarly, nitration of protein could also contribute to disease pathogenesis. To assess the status of lipid peroxidation and/or RONS, autoimmune prone female MRL+/+ mice (5-week old) were treated with trichloroethene (TCE), an environmental contaminant known to induce autoimmune response, for 48 weeks (0.5mg/ml via drinking water), and formation of antibodies to LPDA-protein adducts was followed in the sera of control and TCE-treated mice. TCE treatment led to greater formation of both anti-MDA- and -HNE-protein adduct antibodies and higher serum iNOS and nitrotyrosine levels. The increase in TCE-induced oxidative stress was associated with increases in anti-nuclear-, anti-ssDNA- and anti-dsDNA antibodies. These findings suggest that TCE exposure not only leads to oxidative/nitrosative stress, but is also associated with induction/exacerbation of autoimmune response in MRL+/+ mice. Further interventional studies are needed to establish a causal role of RONS in TCE-mediated autoimmunity. PMID- 17123687 TI - Simplified dispatch-assisted CPR instructions outperform standard protocol. AB - OBJECTIVE: Dispatch-assisted chest compressions only CPR (CC-CPR) has gained widespread acceptance, and recent research suggests that increasing the proportion of compression time during CPR may increase survival from out-of hospital cardiac arrest. We created a simplified CC-CPR protocol to reduce time to start chest compressions and to increase the proportion of time spent delivering chest compressions. This simplified protocol was compared to a published protocol, Medical Priority Dispatch System (MPDS) Version 11.2, recommended by the National Academies of Emergency Dispatch. METHODS: Subjects were randomized to the MPDS v11.2 protocol or a simplified protocol. Data was recorded from a Laerdal Resusci Anne Skillreporter manikin. A simulated emergency medical dispatcher, contacted by cell phone, delivered standardized instructions for both protocols. Outcomes included chest compression rate, depth, hand position, full release, overall proportion of compressions without error, time to start of CPR and total hands-off chest time. Proportions were analyzed by Wilcoxon's Rank Sum tests and time variables with Welch ANOVA and Wilcoxon's Rank Sum test. All tests used a two-sided alpha-level of 0.05. RESULTS: One hundred and seventeen subjects were randomized prospectively, 58 to the standard protocol and 59 to the simplified protocol. The average age of subjects in both groups was 25 years old. For both groups, the compression rate was equivalent (104 simplified versus 94 MPDS, p = 0.13), as was the proportion with total release (1.0 simplified versus 1.0 MPDS, p = 0.09). The proportion to the correct depth was greater in the simplified protocol (0.31 versus 0.03, p < 0.01), as was the proportion of compressions done without error (0.05 versus 0.0, p = 0.16). Time to start of chest compressions and total hands-off chest time were better in the simplified protocol (start time 60.9s versus 78.6s, p < 0.0001; hands-off chest time 69 s versus 95 s, p < 0.0001). The proportion with correct hand position, however, was worse in the simplified protocol (0.35 versus 0.84, p < 0.01). CONCLUSIONS: The simplified protocol was as good as, or better than the MPDS v11.2 protocol in every aspect studied except hand position, and the simplified protocol resulted in significant time savings. The protocol may need modification to ensure correct hand position. Time savings and improved quality of CPR achieved by the new set of instructions could be important in strengthening critical links in the cardiac chain of survival. PMID- 17123688 TI - Gas embolism following intraosseous medication application proven by post-mortem multislice computed tomography and autopsy. AB - The post-mortem use of modern imaging techniques such as multislice computed tomography (MSCT) is becoming increasingly important as an aid for conventional autopsy. This article presents a case of a 4-month-old boy who died from sudden infant death syndrome (SIDS) with intravascular gas after an intraosseus medication application documented by post-mortem MSCT. It is most likely that the gas entered the body during resuscitation. This case emphasises the advantage of post-mortem imaging as a complementary aid for the autopsy. We conclude that during emergency treatment, the medical staff should be aware of the possibility of causing a gas embolism following intraosseus medication. Resuscitation with an inserted, disconnected intraosseous needle should be avoided. PMID- 17123689 TI - Terlipressin as an adjunct vasopressor in refractory hypotension after tricyclic antidepressant intoxication. AB - AIM OF STUDY: To report the management of cardiovascular failure refractory to standard catecholamine therapy with terlipressin in a patient with tricyclic antidepressant (TCA) intoxication. CASE REPORT: A 41-year-old woman, with suicidal ingestion of 11.25 g amitriptyline and 1500 mg diclofenac, was admitted to the emergency department. After 30 min in ventricular fibrillation, with ongoing CPR, she regained a potentially perfusing rhythm, but with hypotension refractory to standard catecholamine therapy with adrenaline, 2 microg/kg/min (norepinephrine); adrenaline, 1 microg/kg/min (epinephrine) until 55 min after admission. An injection of 1 mg terlipressin restored mean arterial blood pressure >65 mmHg within 10 min. Ten hours after admission to the intensive care unit, catecholamine support could be withdrawn because of a stable haemodynamic state. Within 7 days, all organ function recovered, and the patient regained full neurological function. CONCLUSIONS: Successful management of cardiovascular failure with terlipressin after TCA intoxication refractory to catecholamines suggests a potential role for terlipressin as an adjunct vasopressor in severely hypotensive patients. PMID- 17123690 TI - Second trimester amniotic fluid endothelin-1 concentrations and subsequent development of intrauterine growth restriction. AB - OBJECTIVE: The purpose of this prospective study was to record endothelin-1 (ET 1) concentrations in the second trimester amniotic fluid and compare these values in women who developed intrauterine growth restriction (IUGR) later in pregnancy with those with uneventful pregnancies. METHOD: Amniotic fluid was retrieved by amniocentesis from 125 women in the second trimester of pregnancy. The levels of ET1 were measured by a sensitive and specific radioimmunoassay. RESULTS: From the 125 women included in the study 12 had pregnancies that later developed IUGR and 88 had uneventful pregnancies. The ET1 concentration was significantly higher (P<0.005) in women who later developed IUGR than in normal pregnancy (106 pg/ml versus 64.7 pg/ml). CONCLUSION: The amniotic fluid concentration of ET1 is elevated by the second trimester in women who later develop IUGR. PMID- 17123691 TI - Re-laparotomy after cesarean section. AB - OBJECTIVE: To assess the rate, indications, and outcome of re-laparotomy after cesarean section in the early postoperative period. DESIGN: A retrospective observational study during a 121-month period. SETTING: A tertiary care university center. POPULATION: A cohort of 3380 women who underwent cesarean section out of 18,609 parturients. MAIN OUTCOME MEASURES: Incidence of re laparotomy after cesarean section in the early postoperative period. RESULTS: The incidence of re-laparotomy after cesarean section was 0.53% (18/3380). Of these 18 women, 12 (66%) were operated for hemorrhage, 3 (17%) for eventration, and 3 (17%) for formation of intra-abdominal abscess. Hysterectomy was required in one case (5.5%). We had no maternal mortalities. CONCLUSION: Although the incidence of re-laparotomy after cesarean in the early postoperative period is low and the outcome is favorable, several measures must be undertaken to reduce the need for re-laparotomy. PMID- 17123692 TI - TNFA-308G>A polymorphism influences the TNF-alpha response to altered vaginal flora. AB - OBJECTIVE: To investigate the association between a tumor necrosis factor-alpha (TNF-alpha) gene polymorphism, vaginal TNF-alpha level, and microbial flora in pregnant women. METHODS: Vaginal samples from 203 women at 18-22 weeks' gestation were analyzed for microflora. TNFA-308G>A polymorphism was analyzed by polymerase chain reaction and restriction endonuclease analysis and TNF-alpha concentration was determined by ELISA. Outcome data were subsequently obtained. RESULTS: The vaginal TNF-alpha concentration was elevated in TNFA-308A carriers only in the presence of abnormal vaginal flora. A median TNF-alpha level of 10.94 pg/ml in TNFA-308A carriers with bacterial vaginosis (BV) was significantly higher than that of 1.77 pg/ml in TNFA-308A carriers without BV (P=.02), and 1.72 pg/ml in TNF-308G homozygotes with BV (P=.01). CONCLUSION: The TNFA-308G>A polymorphism influences the local TNF-alpha response to altered vaginal microflora. This suggests that the nature of the host response to microbial invasion in the lower female genital is genetically determined. PMID- 17123693 TI - Management of cervical weakness based on the measurement of cervical resistance index. AB - OBJECTIVE: To assess the value of measuring cervical resistance index (CRI) as an aid to selecting patients with a history of spontaneous mid-trimester miscarriage for cervical cerclage in subsequent pregnancies. STUDY DESIGN: An observational study of 175 patients with a history of one or more spontaneous mid-trimester losses and 123 non-pregnant women who had CRI measurements performed while undergoing routine gynaecological surgery. Those women whose CRI indicated an incompetent cervix were recommended for cervical cerclage in future pregnancies while women with a normal CRI were recommended for conservative management without cerclage. RESULTS: The median CRI in the 123 control women was 38.26 N while the median CRI in the study group was 17.00 N. In 62 of the 175 study women (35%) the CRI findings were at variance with the history of previous mid trimester loss; 30 (16.6%) were deemed competent on CRI whereas the history suggested incompetence and 32 (18.4%) were incompetent on CRI while the history suggested that the cervix should be competent. The 175 study women had had 486 previous pregnancies with a successful outcome in 27.4% of the pregnancies. Ninety-four patients have now had 148 pregnancies with a successful outcome in 75.8% of the pregnancies. CONCLUSIONS: Non-pregnant women with a history of spontaneous mid-trimester miscarriage have a significantly lower cervical resistance index than parous women who have not suffered mid-trimester loss. In 35% of patients the CRI was at variance with the history of the previous loss. CRI may be a useful technique to aid the diagnosis of cervical weakness allowing a rational selection for treatment with prophylactic cervical cerclage. PMID- 17123694 TI - French trainees in obstetrics and gynaecology theoretical training and practice of vaginal breech delivery: a national survey. AB - OBJECTIVE: To survey French trainees in Obstetrics and Gynaecology and evaluate their theoretical training and practice of vaginal breech delivery (VBD). STUDY DESIGN: We conducted a national survey between January and April 2004 among the 817 French trainees undergoing a 5-year training program in obstetrics and gynaecology. Trainees in years 1-3 were considered as juniors, and those in years 4 and 5 as seniors. Respondents were invited to specify whether they had received theoretical teaching in the management of VBD, and to indicate the number of VBDs they had performed during their training. RESULTS: The questionnaire was returned by 156 (19%) trainees and of these 140 questionnaires were suitable for analysis. Overall, 35% of the respondents had never been taught the management of a VBD. Among the senior trainees, 33% had performed less than four VBDs, and 23% had not received any teaching on how to manage a VBD. CONCLUSION: The level of theoretical training and practice in the management of VBD seems to have diminished and does not bring the French trainees in obstetrics and gynaecology to the required standard of competence. PMID- 17123695 TI - Vasorelaxant effect of levcromakalim on isolated umbilical arteries of preeclamptic women. AB - OBJECTIVE: Potassium channel openers are revealed to be a new type of antihypertensive drug. We aimed to clarify the effects of levcromakalim, an ATP sensitive potassium channel opener, on human isolated umbilical artery (UA) and to compare them with those of nifedipine and magnesium sulphate, which are currently used in the treatment of preeclampsia (PE). STUDY DESIGN: A total of 52 umbilical arteries, isolated immediately after delivery from 27 healthy and 25 preeclamptic pregnant women, were placed into 10-ml organ baths filled with Kreb's solution at physiological pH and temperature. The concentration-dependent relaxations in response to levcromakalim, nifedipine and magnesium sulphate were compared in vessels precontracted with serotonin (1 micromol/l). RESULTS: The maximal relative relaxation responses (E(max), expressed as percentage of serotonin-induced precontraction) to magnesium sulphate, nifedipine and levcromakalim in umbilical arteries were identical in the healthy (85.06+/-3.31, 84.80+/-3.01 and 80.37+/-5.32%, respectively) and preeclamptic (77.20+/-5.30, 83.36+/-2.37 and 79.13+/-4.30%, respectively) groups. CONCLUSION: Levcromakalim has a vasodilatory effect on the umbilical artery like magnesium sulphate and nifedipine, and serves as an antihypertensive potential that might be used in the treatment of preeclampsia. However, further experimental and clinical studies are needed to propose that ATP-sensitive potassium channel openers are beneficial drugs in cases of clinical preeclampsia. PMID- 17123696 TI - Plasma homocysteine and polycystic ovary syndrome: the missed link. AB - OBJECTIVE: The objective was to investigate the relationship between insulin resistance and increased serum homocysteine in women with polycystic ovarian syndrome (PCOS). DESIGN: Prospective controlled trial. SETTING: Department of Obstetrics & Gynecology, Mansoura Faculty of Medicine, Mansoura, Egypt. PATIENTS: Ninety PCOS women as a study group and 35 women with infertility due to other causes as a control group. OUTCOME MEASURES: Serum homocysteine levels in the presence and absence of insulin resistance in PCOS patients. RESULTS: Homocysteine levels were significantly higher in PCOS patients than in the controls. Considering 11 micromol/l as the cut-off level for a normal homocysteine level, 41.1% of PCOS patients (37 out of 90) and 2.9% of control group (1 out of 35) had high homocysteine levels. With regard to insulin resistance, 23% of PCOS patients without insulin resistance (9 out of 39) had a high homocysteine level, while 47% of PCOS patients with insulin resistance (24 out of 51) had this, thus demonstrating the effect of insulin resistance on the homocysteine level. CONCLUSION: There is a strong association between serum homocysteine and insulin resistance in women with PCOS that contributes to the long-term complications of PCOS. PMID- 17123697 TI - The influence of mode of delivery and ACE genotype on serum angiotensin converting enzyme (ACE) activity in the mother and infant at term. AB - OBJECTIVE: Angiotensin converting enzyme (ACE) and its genotype have been shown to play a role in the pathophysiology of pregnancy complications such as pre eclampsia and intrauterine growth restriction and possibly in adult onset chronic diseases. The physiological changes of ACE and the influence of its genotype during the intrapartum period are not well known. Hence the aim of this study was to assess serum ACE activity and its genotype in mothers and infants at term in relation to labour and mode of delivery. STUDY DESIGN: A cross sectional study of 99 women who laboured and 27 women who delivered by elective caesarean section after 36 completed weeks gestation with uncomplicated pregnancies. Venous cord bloods were obtained immediately after delivery of the placenta for serum ACE activity, ACE genotype and acid-base status. Maternal venous samples were obtained just after delivery for analysis of ACE activity and ACE genotype. Univariate analyses were performed using parametric tests for normally distributed data and nonparametric tests for the data that were not normally distributed. A multiple regression model was developed to adjust for potential confounding factors. RESULTS: The umbilical venous ACE activity was similar for infants delivered following labour compared to those delivered by elective caesarean section, 47.2 U/L (35-64) versus 40.1 U/L (31-60) (adjusted p=0.21). Maternal ACE activities were 28.9 U/L (22-35) and 32.1 U/L (22-40) respectively (adjusted p=0.17). The ACE activity in infants was higher than that of mothers 46 U/L versus 22 U/L, respectively (p= or <0.001). Neither the mode of delivery nor the presence of suspected fetal compromise influenced maternal or infant ACE activity. There was no influence of the infants' genotype on ACE activity in relation to mode of delivery. The DD genotype was associated with higher ACE activity in mothers (p=0.001) but not in infants (p=0.56). CONCLUSIONS: This study shows that intrapartum events do not affect ACE activity. These results will enhance our ability to investigate the role of ACE and its genotype in abnormal fetal growth and in subsequent adult onset chronic disease. PMID- 17123698 TI - Application of a new leaf area index algorithm to China's landmass using MODIS data for carbon cycle research. AB - An operational system was developed for mapping the leaf area index (LAI) for carbon cycle models from the moderate resolution imaging spectroradiometer (MODIS) data. The LAI retrieval algorithm is based on Deng et al. [2006. Algorithm for global leaf area index retrieval using satellite imagery. IEEE Transactions on Geoscience and Remote Sensing, 44, 2219-2229], which uses the 4 scale radiative transfer model [Chen, J.M., Leblancs, 1997. A 4-scale bidirectional reflection model based on canopy architecture. IEEE Transactions on Geoscience and Remote Sensing, 35, 1316-1337] to simulate the relationship of LAI with vegetated surface reflectance measured from space for various spectral bands and solar and view angles. This algorithm has been integrated to the MODISoft platform, a software system designed for processing MODIS data, to generate 250 m, 500 m and 1 km resolution LAI products covering all of China from MODIS MOD02 or MOD09 products. The multi-temporal interpolation method was implemented to remove the residual cloud and other noise in the final LAI product so that it can be directly used in carbon models without further processing. The retrieval uncertainties from land cover data were evaluated using five different data sets available in China. The results showed that mean LAI discrepancies can reach 27%. The current product was also compared with the NASA MODIS MOD15 LAI product to determine the agreement and disagreement of two different product series. LAI values in the MODIS product were found to be 21% larger than those in the new product. These LAI products were compared against ground TRAC measurements in forests in Qilian Mountain and Changbaishan. On average, the new LAI product agrees with the field measurement in Changbaishan within 2%, but the MODIS product is positively biased by about 20%. In Qilian Mountain, where forests are sparse, the new product is lower than field measurements by about 38%, while the MODIS product is larger by about 65%. PMID- 17123699 TI - Social, economic, and ecological impacts of the "Grain for Green" project in China: a preliminary case in Zhangye, Northwest China. AB - This paper provides applications of the integrated assessment (IA) approach in a case study in the Heihe River Basin of Northwest China. Some socio-economic and ecological impact results of forestry land use scenarios are presented in the paper. While seven types of land use scenarios for carbon sequestration purposes were considered for the IA applications, this paper mainly presents impacts of land use scenarios within the Grain for Green (GFG) category [see Yin et al., this volume]. China's national Grain for Green Project was implemented in order to protect and improve ecosystems, while allowing an evolution of agricultural management practices compatible with raising peasants' incomes. Zhangye Prefecture, located in the Heihe River Basin of an arid area in Northwest China, was chosen as the investigation site of the IA case study. Based on fundamental orientation theory, the social sustainability impacts of GFG land use options were assessed. Between 2002 and 2004, the GFG project brought US$23.56 million yuan in net income to the prefecture's peasants. Project implementation resulted in a 1.71 Gg increase in net primary productivity (NPP), as well as a 44.36 Gg rise in net ecosystem productivity (NEP). This suggested that, in Zhangye Prefecture, the Grain for Green Project could enhance the sustainability and stability of the local society, increase peasants' net income, as well as protect and remediate local ecosystems. PMID- 17123700 TI - Spatial scaling between leaf area index maps of different resolutions. AB - We developed algorithms for spatial scaling of leaf area index (LAI) using sub pixel information. The study area is located near Liping County, Guizhou Province, in China. Methods for LAI spatial scaling were investigated on LAI images with 960 m resolution derived in two ways. LAI from distributed calculation (LAID) was derived using Landsat ETM+ data (30 m), and LAI from lumped calculation (LAIL) was obtained from the coarse (960 m) resolution data derived through resampling the ETM+ data. We found that lumped calculations can be considerably biased compared to the distributed (ETM+) case, suggesting that global and regional LAI maps can be biased if surface heterogeneity within the mapping resolution is ignored. Based on these results, we developed algorithms for removing the biases in lumped LAI maps using sub-pixel land cover-type information, and applied these to correct one coarse resolution LAI product which greatly improved its accuracy. PMID- 17123701 TI - Comparison of alternative scoring techniques when assessing decision maker's multi-objective preferences in natural resource management. AB - A popular way to assess a decision maker's preferences in multi-objective natural resource management is to ask the decision maker to compare alternative management plans pairwisely in the ratio scale. Several numerical scoring techniques have been proposed for the ratio scale comparisons, but the performance of the alternative techniques is not fully understood. At the same time, the choice of scoring technique potentially impacts the description of the decision maker's preferences and therefore the actual management decisions. In this paper, the regression model for the ratio scale pairwise comparisons is used, and the differences between the alternative scoring techniques are studied based on two different viewpoints. The first idea is to interpret the scoring techniques as fixed models and then compare them under fixed numerical scaling. The applicability of several potential decision-making strategies is discussed in this context. The second idea is to parameterise the scoring techniques and then compare them under optimal scaling. This was possible in an artificial experiment, where the true values of the alternatives are known. The results supported the use of a geometric scoring technique. Also, the importance of assessing an appropriate scale parameter of the geometric progression is pointed out. PMID- 17123702 TI - On the origin of syn- and metachronous urothelial carcinomas. AB - OBJECTIVE: To evaluate existing models for the origin of meta- and synchronous urothelial carcinomas in light of the accumulated genetic data. METHODS: Published studies on the clonal origin and genetic relationships of syn- and metachronous tumors, genetic aberrations in normal and premalignant urothelial lesions, as well as histologic and genetic mapping studies of cystectomized bladder samples are reviewed. RESULTS: The most common models for the origin of syn- and metachronous tumors are found to conform less well to the accumulated genetic data. A new model is proposed, the field-first-tumor-later model, in which aberrant cells with a stem cell, or stem cell-like, origin spread in the urothelium by cellular displacement, creating fields of premalignant cells. Tumor growth is suggested to be initiated by critical genetic events occurring in individual cells in such fields. Hence, recurring tumors are proposed to originate from a shared field of premalignant cells and not from previous overt tumors. CONCLUSIONS: The proposed model can better account for the existing genetic and histological data on syn- and metachronous urothelial carcinomas. PMID- 17123703 TI - Comparison of codon usage and tRNAs in mitochondrial genomes of Candida species. AB - To gain insight into the nature of the mitochondrial genomes (mtDNA) of different Candida species, the synonymous codon usage bias of mitochondrial protein coding genes and the tRNAs in C. albicans, C. parapsilosis, C. stellata, C. glabrata and the closely related yeast Saccharomyces cerevisiae were analyzed. Common features of the mtDNA in Candida species are a strong A+T pressure on protein coding genes, and insufficient mitochondrial tRNA species are encoded to perform protein synthesis. The wobble site of the anticodon is always U for the NNR (NNA and NNG) codon families, which are dominated by A-ending codons, and always G for the NNY (NNC and NNU) codon families, which is dominated by U-ending codons, and always U for the NNN (NNA, NNU, NNC and NNG) codon families, which are dominated by A ending codons and U-ending codons. Patterns of synonymous codon usage of Candida species can be classified into three groups: (1) optimal codon-anticodon usage, Glu, Lys, Leu (translated by anti-codon UAA), Gln, Arg (translated by anti-codon UCU) and Trp are containing NNR codons. NNA, whose corresponding tRNA is encoded in the mtDNA, is used preferentially. (2) Non-optimal codon-anticodon usage, Cys, Asp, Phe, His, Asn, Ser (translated by anti-codon GCU) and Tyr are containing NNY codons. The NNU codon, whose corresponding tRNA is not encoded in the mtDNA, is used preferentially. (3) Combined codon-anticodon usage, Ala, Gly, Leu (translated by anti-codon UAG), Pro, Ser (translated by anti-codon UGA), Thr and Val are containing NNN codons. NNA (tRNA encoded in the mtDNA) and NNU (tRNA not encoded in the mtDNA) are used preferentially. In conclusion, we propose that in Candida species, codons containing A or U at third position are used preferentially, regardless of whether corresponding tRNAs are encoded in the mtDNA. These results might be useful in understanding the common features of the mtDNA in Candida species and patterns of synonymous codon usage. PMID- 17123704 TI - Demonstration of adiponectin receptors 1 and 2 mRNA expression in human breast cancer cells. AB - Recently, we have shown that low adiponectin levels are significantly associated with an increased breast cancer risk. It seems to be very important to study the expression of adiponectin receptor 1 (AdipoR1) and receptor 2 (AdipoR2) in the human breast epithelial cells and breast cancer cells in order to clarify whether or not adiponectin exerts its effects directly on these cells. Expression of adiponectin, AdipoR1, and AdipoR2 mRNA was determined by RT-PCR assay using the RNA samples obtained from human breast cancer cell lines (MCF-7, T47D, SKBR3, and MDA-MB231), HMEC (primary culture of normal human mammary epithelial cells), adipose tissues (axilla) as well as breast cancer cells and normal breast epithelial cells selectively collected from breast cancer tissues by laser microdissection (LMD). Adiponectin mRNA expression was observed only in the adipose tissues. On the other hand, AdipoR1 and AdipoR2 mRNA expression was observed in all four breast cancer cell lines, HMEC, adipose tissues as well as breast cancer cells and normal breast epithelial cells selectively collected by LMD. In addition, AdipoR1 and AdipoR2 expression in both normal breast epithelial cells and breast cancer cells was confirmed by immunohistochemistry. These results suggest a possibility that adiponectin might modulate the growth of normal breast epithelial cells and breast cancer cells directly through AdipoR1 and AdipoR2 receptors, and that the association of low serum adiponectin levels with a high breast cancer risk might be explained, at least in part, by the direct effect of adiponectin on the breast epithelial cells. PMID- 17123705 TI - Removal of mercury from an alumina refinery aqueous stream. AB - Digestion condensate is formed as a by-product of the alumina refinery digestion process. The solution exhibits a high pH and is chemically reducing, containing many volatile species such as water, volatile organics, ammonia, and mercury. Because digestion condensate is chemically unique, an innovative approach was required to investigate mercury removal. The mercury capacity and adsorption kinetics were investigated using a number of materials including gold, silver and sulphur impregnated silica and a silver impregnated carbon. The results were compared to commercial sorbents, including extruded and powdered virgin activated carbons and a sulphur impregnated mineral. Nano-gold supported on silica (88% removal under batch conditions and 95% removal under flow conditions) and powdered activated carbon (91% under batch conditions and 98% removal under flow conditions) were the most effective materials investigated. The silver and sulphur impregnated materials were unstable in digestion condensate under the test conditions used. PMID- 17123706 TI - Cerebral hypoxia-ischemia and middle cerebral artery occlusion induce expression of the cannabinoid CB2 receptor in the brain. AB - Until recently the cannabinoid CB2 receptor was believed to be absent from the central nervous system. In this study we have identified CB2 expressing cells that appear in the rat brain following stroke and hypoxic-ischemia. At 3 days following surgery CB2-positive macrophages, deriving from resident microglia and/or invading monocytes appear on the lesioned side of the brain. By day 7, a mixed population of CB2-positive cells is present. Microglia-derived macrophages are the key cells in the first stages of brain inflammation, and a pivotal step in the neurodegeneration that follows the acute stage of injury. Thus, CB2 may be important in the brain during injury, and in inflammatory neurodegenerative disorders. The presence of CB2-positive cells in the brain following stroke may provide a novel strategy for cannabinoid-mediated intervention into stroke induced neurodegeneration without the psychoactive effects of CB1 receptor stimulation. PMID- 17123707 TI - Altered extracellular levels of DOPAC and HVA in the rat nucleus accumbens shell in response to sub-chronic nandrolone administration and a subsequent amphetamine challenge. AB - Associated with acts of violence and polydrug use, abuse of anabolic androgenic steroids (AAS) is an increasing problem in society. The aim of the present study was to elucidate whether sub-chronic treatment with the AAS nandrolone decanoate affects dopamine release and dopamine metabolism in the rat nucleus accumbens shell, before and after an amphetamine challenge. Male Sprague-Dawley rats received daily i.m. injections of nandrolone decanoate (15 mg/kg) or vehicle for 14 days. On day 15, the animals were anaesthetized and a microdialysis probe was implanted into the nucleus accumbens shell. Extracellular fluid was collected 1h before and 3h after a single amphetamine injection (5 mg/kg). The samples were then analyzed regarding the content of dopamine, and its metabolites 3,4 dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), using HPLC with electrochemical detection. Two weeks of nandrolone decanoate administration caused a significant decrease of the basal DOPAC and HVA levels, which remained low during the first hour following the amphetamine challenge. Dopamine levels did not differ significantly between groups, neither after the nandrolone pre treatment nor the amphetamine challenge. In conclusion, these novel findings indicate that AAS alter the metabolism of dopamine in a brain region involved in the development of drug dependence. PMID- 17123708 TI - No association between TPH2 gene polymorphisms and ADHD in a UK sample. AB - Tryptophan Hydroxylase 2 (TPH2) is the rate-limiting enzyme in the biosynthesis of serotonin which is exclusively expressed in the brain. Recent molecular studies reported significant association between markers mapped to TPH2 and psychiatric conditions including ADHD. We have examined four single nucleotide polymorphisms (SNPs) two of which (rs1843809, rs1386493) were reported to associate with ADHD in an Irish ADHD sample. Transmission disequilibrium analysis revealed no significant association between any of these markers and ADHD. Dividing by the sex of the transmitting parent has also failed to replicate the previously reported paternal over-transmission of the associated alleles to ADHD probands. A larger sample size will be required to clarify if TPH2 alleles are or are not associated with ADHD. PMID- 17123709 TI - Reduced aspartate release from rat hippocampal synaptosomes loaded with Clostridial toxin light chain by electroporation: evidence for an exocytotic mechanism. AB - Aspartate can be released from certain hippocampal pathways along with glutamate or GABA. Although aspartate immunoreactivity has been localized to synaptic vesicles and aspartate release is Ca(2+)-dependent, there has been no clear evidence favoring an exocytotic mechanism. In particular, pretreatment with Clostridial toxins has not consistently inhibited aspartate release, even when release of glutamate from the same tissue samples was markedly inhibited. To address this issue directly, rat hippocampal synaptosomes were permeabilized transiently by electroporation in the presence of active or inactivated Clostridial toxin light chains. Loading rat hippocampal synaptosomes with the active light chain of tetanus toxin or of botulinum neurotoxins A, B or C reduced the K(+)-evoked release of aspartate at least as much as that of glutamate. These results confirm that aspartate is released by exocytosis in rat hippocampus. PMID- 17123710 TI - Neural correlates of transmeatal cochlear laser (TCL) stimulation in healthy human subjects. AB - Transmeatal cochlear laser (TCL) treatment has recently been proposed as a therapeutic procedure for cochlear dysfunction such as chronic cochlear tinnitus or sensorineural hearing loss. The aim of this study was to investigate whether TLC has any influence on the central nervous system using functional MRI with healthy young adults. The laser stimulation device was placed on the tympanic membrane of both ears. A laser stimulation run and a placebo run were performed in random order. The participants were unable to differentiate between verum and placebo stimulation. In the comparison of verum to placebo runs, we observed significant activations within the left superior frontal gyrus, the right middle and medial frontal gyrus, the right superior parietal lobule, the left superior occipital gyrus, the precuneus and cuneus bilaterally, the right anterior and the left and right middle and posterior cingulate gyrus and the left thalamus. This network of brain areas corresponds well to results from previous PET studies of patients with tinnitus. Though TCL seems to have a clinically measurable effect on the central nervous system the neurophysiological mechanism leading to the observed activated neuronal network remains unknown. PMID- 17123711 TI - Relationships between plasma leptin concentrations and human brain structure: a voxel-based morphometric study. AB - We have previously demonstrated that obese people have reduced grey matter (GM) in several brain areas, including regions implicated in the regulation of taste (i.e., inferior frontal operculum and postcentral gyrus), reward (i.e., putamen), and behavioural processing (i.e., middle frontal gyrus), compared with their lean counterparts. It is well established that the brain may serve as a direct target for adiposity signals, one of the most important being leptin. We investigated the relationships between fasting plasma leptin concentrations and brain tissue composition in a group of 32 young adult Caucasians (12M/20F, age 32+/-1 years, body fat 29+/-1%, mean+/-S.E.) with normal glucose tolerance by using voxel-based morphometry of magnetic resonance imaging scans. Fasting plasma leptin concentrations were positively correlated with GM volumes of the left cerebellum and left inferior temporal gyrus and negatively associated with GM volumes of the left inferior frontal operculum, left postcentral gyrus, and right putamen (P<0.001, uncorrected for multiple comparisons) after adjustment for sex, percent body fat, age, fasting plasma insulin concentrations (i.e., the major determinants of plasma leptin), and global GM volume (thus allowing for an assessment of regional effects only). This study showed an independent, negative correlation between fasting plasma leptin concentrations, which are increased in obesity, and the volumes of GM in brain areas where obese people have reduced GM compared to their lean counterparts. These relationships may explain some of the abnormalities in brain morphology recently found to be associated with excess body fatness. PMID- 17123712 TI - Arginine vasopressin in periaqueductal gray, which relates to antinociception, comes from hypothalamic paraventricular nucleus in the rat. AB - Hypothalamic paraventricular nucleus (PVN) is a major source of arginine vasopressin (AVP). Our previous work has proven that: (1) pain stimulation enhances PVN synthesis and secretion of AVP; (2) AVP in periaqueductal gray (PAG) plays a role in antinociception; (3) pain stimulation increases AVP concentration in PAG tissue. The present study was to investigate AVP source in PAG during pain modulation of the rat. The results showed that: (1) pain stimulation elevated AVP concentration in both PVN and PAG perfusion liquid, in which the peak of AVP concentration in PVN perfusion liquid occurred earlier than that in PAG perfusion liquid; (2) PVN cauterization weakened pain stimulation-induced PAG releasing AVP, in which the inhibitive effect of bilateral PVN cauterization showed stronger than that of unilateral PVN cauterization; (3) microinjection of l glutamate sodium into PVN, which excited local neurons, increased AVP concentration in PAG perfusion liquid in a dose-dependent manner. The data suggest that AVP in PAG, which relates with pain modulation, comes from PVN. PMID- 17123713 TI - Protracted myelin clearance hinders central primary afferent regeneration following dorsal rhizotomy and delayed neurotrophin-3 treatment. AB - Regeneration within or into the CNS is thwarted by glial inhibition at the site of a spinal cord injury and at the dorsal root entry zone (DREZ), respectively. At the DREZ, injured axons and their distal targets are separated by degenerating myelin and an astrocytic glia limitans. The different glial barriers to regeneration following dorsal rhizotomy are temporally and spatially distinct. The more peripheral astrocytic barrier develops first, and is surmountable by neurotrophin-3 (NT-3) treatment; the more central myelin-derived barrier, which prevents dorsal horn re-innervation by NT-3-treated axons, becomes significant only after the onset of myelin degeneration. Here we test the hypothesis that in the presence of NT-3, axonal regeneration is hindered by myelin degeneration products. To do so, we used the Long Evans Shaker (LES) rat, in which oligodendrocytes do not make CNS myelin, but do produce myelin-derived inhibitory proteins. We show that delaying NT-3 treatment for 1 week in normal (LE) rats, while allowing axonal penetration of the glia limitans and growth within degenerating myelin, results in misdirected regeneration with axons curling around presumptive degenerating myelin ovoids within the CNS compartment of the dorsal root. In contrast, delaying NT-3 treatment in LES rats resulted in straighter, centrally-directed regenerating axons. These results indicate that regeneration may be best optimized through a combination of neurotrophin treatment plus complete clearance of myelin debris. PMID- 17123714 TI - Long-term hypergravity induces plastic alterations in vestibulo-cardiovascular reflex in conscious rats. AB - To test the hypothesis that an altered gravitational environment induces plastic changes in the vestibulo-cardiovascular reflex, arterial pressure (AP) and hypothalamic glutamate concentration were examined in 2 groups of conscious rats, i.e., a 3-G group and a 1-G group, in which rats were maintained under a 3-G and 1-G environment for 2 weeks, respectively. The vestibulo-cardiovascular reflex was stimulated by a gravitational change induced by a parabolic flight that consisted of 3 phases: "pull-up", during which the G load gradually increased to 2G; a 20s "push-over" into microgravity; and "pull-out", during which the G load increased to 1.8. In the 1-G group, the AP increased by 11.9+/-1.2 mmHg during the pull-up hypergravity period. The AP response was significantly attenuated in the 3-G group (4.0+/-0.8 mmHg). During the push-over microgravity period, the AP decreased from the peak level in the pull-up period and recovered to the pre parabolic control level (-1.8+/-2.4 mmHg). In rats of the 3-G group, the AP was not altered by push-over microgravity. These AP responses were associated with a significant increase in the glutamate concentration in the hypothalamus (4.4+/ 0.7%). The glutamate response was also significantly attenuated in the 3-G group compared with that in the 1-G group. These results indicate that an altered gravitational environment induces plastic alterations in the vestibulo cardiovascular reflex. PMID- 17123715 TI - Combined neuroprotective effects of celecoxib and memantine in experimental intracerebral hemorrhage. AB - Memantine, a N-methyl-D-aspartate (NMDA) receptor antagonist, inhibits hematoma expansion and celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, reduces perihematomal inflammation in intracerebral hemorrhage. We examined whether the combination treatment has additive effects in experimental intracerebral hemorrhage (ICH). ICH was induced using stereotaxic infusion of collagenase into brains of adult rats. After the induction of ICH, rats were treated with intraperitoneal injection of memantine (20 mg/kg), celecoxib (20 mg/kg) or both agents. Only vehicles were administrated in rats of the control group. Results showed that the combination treatment of memantine and celecoxib reduced both hematoma volume and brain edema. Combination treatment also induced the better functional recovery with further attenuation of cerebral inflammation and apoptosis compared to the control group. When compared to the single agent groups, the combination treatment showed better effects in neuroprotection and anti-inflammation. These results suggest the feasible combined application of memantine and celecoxib in ICH treatment. PMID- 17123716 TI - Neurotransmitter depletion by bafilomycin is promoted by vesicle turnover. AB - Accumulation of neurotransmitter into synaptic vesicles is powered by the vacuolar proton ATPase. We show here that, in brain slices, application of the H(+)-ATPase inhibitors bafilomycin or concanamycin does not efficiently deplete glutamatergic vesicles of transmitter unless vesicle turnover is increased. Simulations of vesicle energetics suggest either that bafilomycin and concanamycin act on the H(+)-ATPase from inside the vesicle, or that the vesicle membrane potential is maintained after the H(+)-ATPase is inhibited. PMID- 17123717 TI - Yohimbine prevents morphine-induced changes of glial fibrillary acidic protein in brainstem and alpha2-adrenoceptor gene expression in hippocampus. AB - The alpha(2)-adrenoceptor antagonist yohimbine is known to oppose to several pharmacological effects of opioid drugs, but the consequences and the mechanisms involved remain to be clearly established. In the present study we have checked the effects of yohimbine on morphine-induced alterations of the expression of key proteins (glial fibrillary acidic protein, GFAP) and genes (alpha(2) adrenoceptors) in rat brain areas known to be relevant in opioid dependence, addiction and individual vulnerability to drug abuse. Rats were treated with morphine in the presence or absence of yohimbine. The effects of the treatments on GFAP expression were studied by immunohistochemical staining in Locus Coeruleus (LC) and Nucleus of the Solitary Tract (NST), two important noradrenergic nuclei. In addition, drug effects on alpha(2)-adrenoceptor gene expression were determined by real time RT-PCR in the hippocampus, a brain area that receives noradrenergic input from the brainstem. Morphine administration increased GFAP expression both in LC and NST as it was previously reported in other brain areas. Yohimbine was found to efficiently prevent morphine-induced GFAP upregulation. Chronic (but not acute) morphine downregulated mRNA levels of alpha(2A)- and alpha(2C)-adrenoceptors in the hippocampus, while simultaneously increased the expression of the alpha(2B)-adrenoceptor gene. Again, yohimbine was able to prevent morphine-induced changes in the levels of expression of the three alpha(2)-adrenoceptor genes. These results correlate the well-established reduction of opioid dependence and addiction by yohimbine and suggest that this drug could interfere with the neural plasticity induced by chronic morphine in central noradrenergic pathways. PMID- 17123718 TI - Unintended interpersonal co-ordination: "can we march to the beat of our own drum?". AB - The main purpose of this article was to identify whether an interpersonal motor co-ordination emerges between two participants when they intentionally tried to not co-ordinate their movements between each other. To do this we conjointly examined both the emergence of the motor co-ordination (collective properties) between the two participants, and the maintenance of the individual intrinsic motor properties present in the interpersonal co-ordination. Six pairs of participants performed three situations where they freely moved their right forearm with no instructions (nor restrictions) on frequency or relative phase. The goal of the first two situations was for participants to intentionally co ordinate (P-on) or not co-ordinate (P-off) their movements between each other. In order to determine his/her individual motor properties, each participant performed this task alone, in a third condition. The cross-wavelet transform (CWT) revealed in the P-off condition the emergence of an unintended co ordination in the frequency domain. Moreover, random distribution of the relative phase appeared, in this condition, whereas in the P-on condition, we observed a collective organization especially at a relative phase around 0 degrees . Conjointly, the wavelet transform (WT) showed the presence of individual intrinsic motor properties (motor signature) in the three conditions. These results indicate that, when there was information sharing, participants could not avoid (unintentionally) coordinating with someone. This study suggests that, in daily life, interactions with other people lead the individual motor behavior to adapt unintended motor co-ordination. PMID- 17123719 TI - Distribution of Notch1-expressing cells and proliferating cells in mouse vomeronasal organ. AB - Vomeronasal receptor neurons (VRNs) proliferate and differentiate continuously in the vomeronasal organ (VNO) throughout life. In adult mice, new VRNs are generated mainly in the marginal region, located in the boundary region between sensory and nonsensory epithelia. The Notch signaling pathway is involved in differentiation in the developing nervous system. To understand the Notch signaling pathway involved in generating VRNs, we focused on the relationship between the expression pattern of Notch1 and the localization of proliferating cells in both developing and regenerating mice VNO, and examined the Notch signaling pathway involved in the development of VNO by in situ hybridization of Notch1 and immunocytochemistry of 5-bromo-2'-deoxyuridine. During embryonic and neonatal development, proliferating cells and Notch1-expressing (+) cells were observed evenly throughout VNO. A large number of proliferating cells and Notch1 (+) cells were observed in embryonic VNO, but gradually decreased during development. The localization of proliferating cells was similar to that of Notch1 (+) cells at each developmental stage. In adult VNO, there are a few proliferating cells and Notch1 (+) cells, which were only in the marginal region of VNO. Seven days after removal of the accessory olfactory bulb (AOB), VRNs proliferated throughout VNO. Although the number of Notch1 (+) cells also increased in VNO, the majority of these were concentrated in the dorsal region of VNO, suggesting that it has two types of differentiating cell. These results suggest that Notch1 plays a role in the differentiation of VRNs during development and regeneration of VRNs after removal of AOB. PMID- 17123720 TI - A new gain-of-function allele in chimpanzee tryptophan hydroxylase 2 and the comparison of its enzyme activity with that in humans and rats. AB - Tryptophan hydroxylase 2 (TPH2) is a rate-limiting enzyme of neuronal serotonin biosynthesis. Recently, two single nucleotide polymorphisms (SNPs) at the exon 11 coding region that resulted in amino acid substitutions in the C-terminal domain have been reported to affect enzyme activity in humans and mice. We determined 175 base-pair sequences of the exon 11 region in nine primate species from all recognized lineages. All nucleotide sequence substitutions were synonymous, with the exception of one adenine (A) to guanine (G) substitution at the 1404th position in the open reading frame (ORF). This substitution leads to a glutamine (Q) to arginine (R) amino acid substitution at the 468th position within chimpanzee sequences. The frequency of the G allele was 0.24 among 66 chimpanzees. Therefore, it is a novel SNP observed in chimpanzees, and we have named these two alleles as ch468Q and ch468R, respectively. When expressed in HeLa cells, ch468R caused an approximate 20% increase in enzyme function during L 5-hydroxytryptophan (5HTP) production (P<0.001). We also surveyed the interspecies difference in enzyme activity among human, chimpanzee, and rat. Although the rat showed an identical amino acid sequence at the C-terminal region as those of human and ch468Q, the rat enzyme was more active than those of human or chimpanzee (P<0.001), indicating the importance of substitutions in other regions. Our findings on the chimpanzee SNP will be a useful genetic marker in understanding the individual difference in the serotonin-related behavior. PMID- 17123721 TI - Increase of acetylcholine release by Panax ginseng root enhances insulin secretion in Wistar rats. AB - The present study was designed to ascertain the effect of Panax ginseng root on plasma glucose and investigate the possible mechanisms for the effect. Ninety minutes after the oral administration of P. ginseng root to fasting Wistar rats, plasma glucose decreased in a dose-dependent manner. Simultaneous with the reduction in plasma glucose, an increase in the plasma level of insulin and C peptide was also observed. Moreover, disruption of the available synaptic acetylcholine (ACh), using the inhibitor for choline uptake (hemicholinium-3), or the inhibitor for vesicular choline transport (vesamicol), abolished the metabolic actions of P. ginseng root. Conversely, physostigmine, at a concentration sufficient to inhibit acetylcholinesterase, enhanced the metabolic effect of P. ginseng root. It is possible that P. ginseng root mediates the release of ACh from nerve terminals to enhance insulin secretion. Blockade of the actions of P. ginseng root by 4-diphenylacetoxy-N-methylpiperdine methiodide (4 DAMP) suggested that the site of action is the muscarinic M(3) receptor. Taken together, the results suggest that P. ginseng root has the ability to increase the release of ACh from nerve terminals in rats so as to stimulate muscarinic M(3) receptors activity located in the pancreatic cells for the secretion of insulin, which in turn lower plasma glucose. PMID- 17123722 TI - Genes encoding for AP-2beta and the Serotonin Transporter are associated with the Personality Character Spiritual Acceptance. AB - In several twin studies the relative contribution of genetic factors for personality traits has amounted to figures between 40 and 60%. In the present study we investigated to which degree polymorphisms in the 5-HTT and AP-2beta genes are implicated in the neural processes involved in the formation of Temperament and Character traits, as estimated by Cloninger's TCI. Considering the background of previous reports, associations with the character Self Transcendence and its sub-scale Spiritual Acceptance in particular, were of interest. A stratified random sample of 200 individuals (total population=5173), matched for age, gender and risk behaviors, from volunteering 16- and 19-year-old adolescents students in Sweden was investigated. Cloninger's TCI inventory was used for investigation of temperament and character traits. Blood samples were used for analyses of a promoter serotonin transporter polymorphism (5-HTTLPR) and an intron 2 polymorphism in the transcription factor AP-2beta gene. Among boys individuals with presence of the short 5-HTTLPR genotype showed lower scores, whereas individuals with presence of the short AP-2beta genotype showed higher scores of personality character Self-Transcendence and its sub-scale Spiritual Acceptance. Among girls no effect of either genotype was found. Both among boys and girls, significant interactive effects were found between 5-HTTLPR and AP 2beta genotypes, with regard to Self-Transcendence and Spiritual acceptance. Boys and girls with the combination of presence of the short 5-HTTLPR, and homozygosity for the long AP-2beta genotype scored significantly lower on Self Transcendence and Spiritual Acceptance. PMID- 17123723 TI - Association study of putative promoter polymorphisms in the neuroplastin gene and schizophrenia. AB - A previous study revealed a number of methamphetamine (METH) and phencyclidine (PCP)-reactive tags in a rat brain through serial analysis of gene expression. The present study extends this previous study by investigating whether two genes, which deduced from METH/PCP-reactive tags, were identified as those encoding human transmembrane proteins of the immunoglobulin (Ig) superfamily, neuroplastin (NPTN) and basigin (BSG), confer genetic susceptibility to schizophrenia by analyzing single nucleotide polymorphisms (SNPs). There were nominally significant differences between the two groups in their allelic frequencies (T Ins/Del, chi2=4.910, d.f.=1, P=0.040) and genotypic distributions (T/T or T/Del, chi2=5.116, d.f.=1, P=0.036) of rs3840846 in the 5'-upstream of NPTN. The two groups differed significantly also in their allelic frequencies (G/T, chi2=4.229, d.f.=1, P=0.044), but not genotypic distributions of rs3743500 in the 5'-upstream of NPTN. The haplotypes constructed from the three SNPs (rs3840846, rs3826047 and rs3743500, in order) in the 5'-upstream of NPTN showed a significant association with schizophrenia (permutation P=0.036), in that T-G-T (permutation P=0.028) and del-G-G (permutation P=0.040) were under-represented and over-represented, respectively, in schizophrenia. A reporter construct driven by the 5'-upstream region containing any haplotype consisting of the three SNPs had substantial transcriptional activity. Notably, a reporter construct containing a haplotype T G-T had significantly lower transcriptional activity as compared with one having a haplotype T-G-G or T-A-G. There was no significant difference between the two groups regarding allelic frequencies, genotypic distribution or the adopted SNP combinatory haplotype for BSG. These results suggest that NPTN may be involved in genetic susceptibility to schizophrenia. PMID- 17123724 TI - Neural basis of redundancy effects in visual object categorization. AB - Bilateral redundant information often leads to improved detection performance compared to only unilateral input. This experiment investigates redundancy effects at the level of object processing in behavioral and functional imaging data. Subjects received different combinations of categorical information presented to both visual hemifields simultaneously. For pictures of faces as well as for pictures of buildings, response latencies were faster when two pictures from the same object category were presented compared to two pictures from different categories. This behavioral advantage was accompanied by increased activation in the respective object-selective areas, i.e. in the Fusiform Face Area (FFA) for pictures of faces and in the Parahippocampal Place Areas (PPAs) for pictures of buildings. These results suggest an involvement of visual object selective areas in the behavioral redundancy gain. PMID- 17123725 TI - Comparison between coated vs. uncoated suture middle cerebral artery occlusion in the rat as assessed by perfusion/diffusion weighted imaging. AB - Differences among models in the temporal evolution of ischemia after middle cerebral artery occlusion (MCAO) in rats may considerably influence the results of experimental treatment studies. Using diffusion and perfusion imaging, we compared the spatiotemporal evolution of ischemia in Sprague-Dawley rats after permanent MCAO (pMCAO) with different types of sutures. Male Sprague-Dawley rats were randomly assigned to pMCAO produced with either 4-0 silicone coated (n=8), or 3-0 uncoated monofilaments (n=8). Serial determination of quantitative cerebral blood flow (CBF) and apparent diffusion coefficient (ADC) maps were performed up to 3 h after pMCAO. Lesion volumes were calculated by using previously validated thresholds and correlated with infarct volume corrected for edema defined by 2,3,5-triphenyltetrazolium chloride (TTC) staining at 24 h after MCAO. The ADC/CBF-defined mismatch volume in the 4-0 coated suture model was present significantly longer (up to 120 min) compared to the uncoated 3-0 suture model (30 min). The TTC-derived infarct volume was significantly larger in the coated model (290.3+/-32.8 mm(3)) relative to the uncoated model (252.3+/-34.6 mm(3)). This study demonstrates that the type of suture may significantly influence the spatiotemporal evolution of the ADC/CBF-mismatch as well as the final infarct volume. These inter-model variations must be taken into account when assessing new therapeutic approaches on ischemic lesion evolution in the rat MCAO model. PMID- 17123726 TI - Force development during target-directed isometric force production in Parkinson's disease. AB - Performing various daily activities requires precise application and control of forces, which has been well addressed in neurologically healthy individuals. Recent experiments have demonstrated that in young, normal subjects generating rapid force pulses over various force amplitudes was accomplished by linearly increasing the rate of force development while keeping time to peak force approximately constant (i.e., a pulse-height control strategy). Using Parkinson's disease (PD) patients the present study examined whether PD patients use a pulse height control strategy during rapid and accurate isometric force production. Subjects were instructed to produce force pulses to three different target amplitudes (15, 35, and 55% of their maximal voluntary contractions) at their preferred speed and as fast as possible. When the task was performed as fast as possible, PD patients differed from controls by producing reduced rates of force development and prolonged times to peak force as a function of force amplitude. During performance of the task at preferred speed, which leaves the rate of force production unconstrained, PD patients did not show improved regulation of time to peak force in scaling different force amplitudes compared to controls. These results suggest that PD patients have a difficulty in utilizing a pulse-height control strategy and that such impairments are not dependent on speed. PMID- 17123727 TI - Interleukin-18 mediated inhibition of LTP in the rat dentate gyrus is attenuated in the presence of mGluR antagonists. AB - Pro-inflammatory cytokines are known to be elevated in several neuropathological states that are associated with learning and memory impairments. We have previously demonstrated the inhibition of long-term potentiation (LTP), a recognised model for memory, in the dentate gyrus region of the rat hippocampus, by interleukin-18. We have also previously shown that the inhibitory effect of TNF-alpha on LTP can be attenuated by inhibitors of metabotropic glutamate receptors (mGluRs). We therefore went on to investigate the effects of the mGluR antagonists MPEP and MTPG on the effect of IL-18 on LTP in the rat dentate gyrus in vitro. Recordings of field excitatory post-synaptic potentials (EPSPs) were made from the medial perforant path of rat hippocampal slices. IL-18 (100 ng/ml) applied for 20 min before-HFS had no significant effect on baseline EPSPs but significantly impaired LTP (IL-18 LTP 116+/-9%, versus control LTP 163+/-6% 1h post-tetanus, P<0.001, n=5). Perfusion of the mGluR5 specific antagonist MPEP (5 microM) for 40 min prior to application of IL-18 had no significant effect on baseline EPSPs but significantly attenuated the inhibitory effect of IL-18 on LTP at 30 min but not 1h (177+/-2% and 138+/-8%, respectively, compared to controls; n=5). Perfusion of the group II mGluR antagonist MTPG (50 microM) for 40 min prior to application of IL-18 had no significant effect on baseline EPSPs but was found to significantly reverse the inhibitory effect of IL-18 on LTP at 1h (164+/ 6% compared to IL-18 alone, n=5). This study provides novel evidence of the involvement of mGluRs in the IL-18 mediated inhibition of LTP. PMID- 17123728 TI - Association study of tryptophan hydroxylase 2 gene polymorphisms in panic disorder. AB - Experimental studies on serotonin (5-HT) availability suggest a role for 5-HT synthesis rate in panicogenesis. Recently, it has been discovered that the tryptophan hydroxylase gene isoform 2 (TPH2), rather than TPH1, is preferentially expressed in the neuronal tissue and, therefore, is primarily responsible for the regulation of brain 5-HT synthesis. In the present case-control genetic association study we investigated whether panic disorder (PD) phenotypes are related to two single nucleotide polymorphisms (SNP) of TPH2, rs1386494 A/G and rs1386483 C/T. The study sample consisted of 213 (163 females and 50 males) PD patients with or without affective comorbidity and 303 (212 females and 91 males) matched healthy control subjects. The allelic and genotypic analyses in the total sample did not demonstrate significant association of PD with the studied SNPs, suggesting that these polymorphisms may not play a robust role in predisposition to PD. However, an association with rs1386494 SNP was observed in the subgroup of female patients with pure PD phenotype, indicating a possible gender-specific effect of TPH2 gene variants in PD. PMID- 17123729 TI - Absence of linear correlation between fluctuations in area of simultaneous recorded monosynaptic responses and Hoffmann's reflexes in the rat. AB - In this study we analyze the possible relationship between fluctuations in area of monosynaptic reflex responses (MSR) and Hoffmann's reflex (H reflexes) in the plantar closed loop pathway of the anesthetized rat. These reflexes were evoked by low-frequency stimuli applied to the sciatic nerve or lateral plantar nerve and then concurrently recorded on the distal tibial nerve or lateral plantar nerve, respectively as well as the lateral plantar muscles in the foot of the anesthetized rat. From trial to trial, H reflexes showed higher variability in area than MSR, whether the latter was recorded in the distal tibial nerve (n=8 experiments) or in the lateral plantar nerve (n=5 experiments). No linear correlation was found between changes in area of concurrently evoked MSR and H reflexes (r(MSR-H,n=8)=0.11+/-0.03 and r(MSR-H,n=5)=0.08+/-0.09, respectively). These findings suggest that trial-to-trial fluctuations in area of H reflexes may involve interaction of several sources of variation, among others to MSR variability (due to pre-, and post-synaptic factors influencing the excitability of spinal motoneurons) in combination with those related to peripheral mechanisms, such as trial to trial activation of a different number of muscle fibers, either by the probabilistic transmitter release from neuromuscular junctions, by activation of motor units of variable size or to fluctuations in excitability of muscle fibers. PMID- 17123730 TI - The influence of low-frequency rTMS on EEG of rats. AB - This study aimed to determine the effect of low-frequency repetitive transcranial magnetic stimulation (rTMS) on electroencephalograms (EEGs) of rats. Fifteen Sprague-Dawley rats were subject to 100 pulses of 0.5 Hz rTMS, or sham stimulation. EEGs were recorded before stimulation and within 1 min after rTMS or sham stimulation. Estimates of the EEG correlation dimension (D(2)) and power spectra were calculated. Results show that the D(2) reduced significantly after low-frequency rTMS, but not after sham stimulation. Mean absolute power (MAP) of the gamma band and relative power (RP) of the beta and gamma bands reduce markedly after low-frequency rTMS, but there are no changes with sham stimulation. These results indicate that low-frequency rTMS could affect cortical activities significantly, but effects were markedly different from those of high frequency rTMS. PMID- 17123731 TI - Spinal NK-1 receptor expressing neurons mediate opioid-induced hyperalgesia and antinociceptive tolerance via activation of descending pathways. AB - Opioids can induce hyperalgesia in humans and in animals. Mechanisms of opiate induced hyperalgesia and possibly of spinal antinociceptive tolerance may be linked to pronociceptive adaptations occurring at multiple levels of the nervous system including activation of descending facilitatory influences from the brainstem, spinal neuroplasticity, and changes in primary afferent fibers. Here, the role of NK-1 receptor expressing cells in the spinal dorsal horn in morphine induced hyperalgesia and spinal antinociceptive tolerance was assessed by ablating these cells with intrathecal injection of SP-saporin (SP-SAP). Ablation of NK-1 receptor expressing cells prevented (a) morphine-induced thermal and mechanical hypersensitivity, (b) increased touch-evoked spinal FOS expression, (c) upregulation of spinal dynorphin content and (d) the rightward displacement of the spinal morphine antinociceptive dose-response curve (i.e., tolerance). Morphine-induced hyperalgesia and antinociceptive tolerance were also blocked by spinal administration of ondansetron, a serotonergic receptor antagonist. Thus, NK-1 receptor expressing neurons play a critical role in sustained morphine induced neuroplastic changes which underlie spinal excitability reflected as thermal and tactile hypersensitivity to peripheral stimuli, and to reduced antinociceptive actions of spinal morphine (i.e., antinociceptive tolerance). Ablation of these cells likely eliminates the ascending limb of a spinal bulbospinal loop that engages descending facilitation and elicits subsequent spinal neuroplasticity. The data may provide a basis for understanding mechanisms of prolonged pain which can occur in the absence of tissue injury. PMID- 17123732 TI - Modality-specific sensory changes in humans after the induction of long-term potentiation (LTP) in cutaneous nociceptive pathways. AB - The impact of long-term potentiation (LTP) in nociceptive pathways on somatosensory perception was examined by means of quantitative sensory testing (QST) in the ventral forearm of 12 healthy human subjects. Electrical high frequency stimulation of the forearm skin (HFS; 5 x 1 s at 100 Hz and 10 x detection threshold) led to an abrupt increase of pain to single electrical test stimuli, which were applied through the same electrode (perceptual LTP +72%, p<0.01). Perceptual LTP outlasted the 1-h observation period. The effects of HFS on somatosensory perception of natural test stimuli in the conditioned skin area were restricted to mechanical submodalities. Subjects exhibited a significant decrease of pain threshold and an increase of pain ratings to suprathreshold pinprick stimuli (p<0.01). In 5 out of 12 subjects (42%) light tactile stimuli led to painful sensations (dynamic mechanical allodynia). Furthermore, a small but significant decrease of threshold to blunt pressure stimuli (p<0.05) was found. In contrast, all thermal modalities comprising cold and warm detection thresholds, cold and heat pain thresholds as well as pain summation (perceptual wind up) remained unaltered. These data show that HFS of peptidergic cutaneous C fiber afferents predominantly modulates Adelta- and Abeta-fiber mediated somatosensory functions, suggesting that LTP in nociceptive pathways enhances human pain sensitivity via interaction of two afferent pathways (extrinsic sensitization). PMID- 17123733 TI - Comment on Nicholas and Asghari: Pain 2006;124:269-79. PMID- 17123734 TI - New evidence for the involvement of spinal fractalkine receptor in pain facilitation and spinal glial activation in rat model of monoarthritis. AB - Fractalkine, a chemokine binding to only one known receptor CX3CR1, has recently been proposed to be a neuron-to-glia signal in the spinal cord leading to microglial activation and glially dependent pain facilitation. The previous studies explored that blockade of endogenous fractalkine, using anti-CX3CR1 neutralizing antibody, dose-dependently attenuated neuropathic pain. The present study examined the role of endogenous fractalkine in inflammatory pain. Intra articular injection of complete Freund's adjuvant (CFA)-induced rat ankle joint monoarthritis (MA) model was used. Western blot analysis revealed that CX3CR1 expression in the spinal cord was significantly increased following CFA-induced MA. Intrathecal injection of anti-CX3CR1 neutralizing antibody both delayed the development of mechanical allodynia and thermal hyperalgesia, and reversed established pain facilitation. Furthermore, blockade of CX3CR1 significantly suppressed activation of spinal glia, especially microglia, evoked by MA. These data provided new evidence for the contribution of endogenous fractalkine to the initiation and early maintenance of inflammatory pain facilitation via activating spinal microglia. PMID- 17123735 TI - Increased plasma substance P and CGRP levels, and high ACE activity in migraineurs during headache-free periods. AB - Substance P (SP), calcitonin gene-related peptide (CGRP), and angiotensin converting enzyme (ACE) may have roles in trigeminovascular nociceptive mechanisms. We investigated interictal levels of SP, CGRP, ACE activity, and their correlation, in a sample of migraineurs. Forty-one patients suffering from migraine with aura (MA), 54 without aura (MO), and 52 non-headache subjects (controls) participated in this study. Blood samples were collected from cubital veins. Plasma levels of SP and CGRP were measured by enzyme immunoassay. Plasma ACE activities were measured spectrophotometrically. SP levels in MA (6.6+/-3.7 pg/ml; mean+/-SD) and MO (6.6+/-3.2 pg/ml) were significantly higher than in controls (4.8+/-2.4 pg/ml) (P<0.01). CGRP levels in MA (18.8+/-8.8 pg/ml) and MO (19.1+/-9.4 pg/ml) were also significantly higher than in controls (13.4+/-4.4 pg/ml) (P<0.01). ACE activities in MA (34.6+/-19.0 U/l) were significantly higher than in MO (25.3+/-13.2 U/l) and controls (27.0+/-20.4 U/l) (P<0.05). There was a significant correlation between SP and CGRP levels (P<0.05). In MA, SP and CGRP showed a tendency toward positive correlation, which was not significant. There was a weak, but significant positive correlation between SP levels and ACE activities (P<0.01). However, a relationship between ACE activities and CGRP levels was not observed. The data suggest that SP, CGRP, and ACE are relevant to migraine pathophysiology, and that they may interact. PMID- 17123736 TI - An algorithm for refinement of lattice parameters using CBED patterns. AB - A new algorithm for calculation of lattice parameters from convergent beam electron diffraction (CBED) patterns has been developed. Like most of the previous approaches to the problem, it is an optimization procedure matching geometric elements of high order Laue zone (HOLZ) lines in experimental patterns to corresponding elements of kinematically simulated patterns. The procedure uses an original objective function based directly on the underlying algebraic equation of the HOLZ lines. Although the new approach requires crystal orientation parameters to be fitted alongside the strain components, it is easier to implement than methods used previously. It is also straightforward to apply to strain determination from multiple patterns. Numerical tests on dynamically simulated patterns show that in the case of one or two patterns, the new procedure gives results that are more reliable than the established method based on HOLZ line intersections. As an example application, the a and c parameters of a TiAl alloy are determined. PMID- 17123737 TI - Occurrence of Cryptosporidium and Giardia in suckling piglets in Norway. AB - Faecal samples from 684 litters of suckling piglets from 100 indoor swine herds from all regions of Norway were examined for the presence of Cryptosporidium and Giardia, using sucrose gradient flotation concentration and immunofluorescent staining. Thirty-one (31%) herds and 57 (8.3%) litters tested positive for Cryptosporidium, while 10 (1.5%) litters in 10 different herds (10%) tested positive for Giardia. Molecular characterisation of nine Cryptosporidium isolates demonstrated both C. suis and Cryptosporidium sp. pig genotype II. There was significantly more diarrhoea among the Cryptosporidium positive piglets than among the Cryptosporidium negative piglets. Diarrhoea was not observed amongst litters in which Cryptosporidium sp. pig genotype II was found. There was a significant difference in the Cryptosporidium prevalence between litters from different geographical areas of Norway. This study demonstrates that both Cryptosporidium (C. suis and Cryptosporidium sp. pig genotype II) and Giardia infections are prevalent among suckling piglets in Norway. PMID- 17123738 TI - Mechanistic insights into linear polyethylenimine-mediated gene transfer. AB - We recently debuted a variety of linear polyethylenimines (LPEIs) with low molecular weight as carriers for gene delivery. The highest transfection efficiency (approximately 44%) was obtained with LPEI 6.6 kDa, while the cytotoxicity remained low (approximately 90% of CHO-K1 cells survived the transfection procedure). Here, we investigated various steps during the transfection process using LPEI 8.1, 5.0 and 1.8 kDa, in order to gain a more complete insight into LPEI-mediated gene transfer and to explore conceptual aspects for further optimization. The cellular uptake characterized by flow cytometry was similar for LPEI 8.1 and 5.0 kDa, while it was significantly lower for LPEI 1.8 kDa. The transfection efficacy in contrast was at NP 24 20.07% for LPEI 8.1 kDa and 39.71% for LPEI 5.0 kDa. This suggests that the endocytosis seems not to be a decisive parameter that determines the efficacy of a polymer in the transfection process. Real-time PCR investigations revealed that LPEI 1.8 kDa likewise or even better protected plasmid from degradation compared to LPEI 5.0 or 8.1 kDa. Furthermore, we found that 1/6 to 1/3 intact plasmid DNA reached the intracellular compartments after complexation with LPEI 1.8 kDa. Therefore, the amount of plasmid DNA available in the cytoplasm seems not to be a limiting factor in the transfection process. That LPEI 8.1-polyplexes built at NP 12 in glucose and transfected in serum-free culture conditions were superior to those built in sodium chloride or transfected in serum-containing conditions points at the structure as a decisive parameter deserving more attention in future studies. PMID- 17123739 TI - Structural analysis of cytochromes P450 shows differences in flexibility of heme 2- and 4-vinyls. AB - Structural analysis of the orientations of heme vinyl side chains was carried out using the published crystallographic data for different cytochromes P450. Torsional angles (tau, C(alpha)C(beta)-C(a)C(b)) show different distributions for the vinyls in positions 2 and 4. Whereas the orientation of 2-vinyls is rather restricted (tau between -120 degrees and -180 degrees ), the 4-vinyls have a much higher mobility over almost the entire conformational space. On the basis of the empirical correlation recently reported for peroxidases (M.P. Marzocchi, G. Smulevich, Relationship between heme vinyl conformation and the protein matrix in peroxidases, J. Raman Spectrosc. 34 (2003), 725-736), an attempt has been made to compare the observed vinyl orientations with the experimental frequencies of the vinyl stretching vibrational modes. The data for P450 proteins do not exactly match the peroxidase-derived function, although a qualitatively similar relationship is likely to exist. Differences between P450 forms suggest a variability in heme-region flexibility and in communication with the rest of enzyme. PMID- 17123740 TI - Postnatal alterations in dopaminergic markers in the human prefrontal cortex. AB - Dopamine in the prefrontal cortex plays a critical role in normal cognition throughout the lifespan and has been implicated in the pathophysiology of neuropsychiatric disorders such as schizophrenia and attention deficit disorder. Little is known, however, about the postnatal development of the dopaminergic system in the human prefrontal cortex. In this study, we examined pre- and post synaptic markers of the dopaminergic system in postmortem tissue specimens from 37 individuals ranging in age from 2 months to 86 years. We measured the levels of tyrosine hydroxylase, the rate limiting enzyme in dopamine biosynthesis, using Western immunoblotting. We also examined the gene expression of the three most abundant dopamine receptors (DARs) in the human prefrontal cortex: DAR1, DAR2 and DAR4, by in situ hybridization. We found that tyrosine hydroxylase concentrations and DAR2 mRNA levels were highest in the cortex of neonates. In contrast, the gene expression of DAR1 was highest in adolescents and young adults. No significant changes across age groups were detected in mRNA levels of DAR4. Both DAR1 and DAR2 mRNA were significantly lower in the aged cortex. Taken together, our data suggest dynamic changes in markers of the dopamine system in the human frontal cortex during postnatal development at both pre-and post-synaptic sites. The peak in DAR1 mRNA levels around adolescence/early adulthood may be of particular relevance to neuropsychiatric disorders such as schizophrenia in which symptoms manifest during the same developmental period. PMID- 17123741 TI - Stress in pregnancy and infant HPA axis function: conceptual and methodological issues relating to the use of salivary cortisol as an outcome measure. AB - Problems regulating behaviour and emotions in infancy may be a risk factor for the development of psychopathology later in life. Compelling evidence from animal models suggests that one potential pathway to early dysregulation is fetal programming of the hypothalamic-pituitary-adrenal (HPA) axis. According to this model, prenatal maternal stress and anxiety during sensitive periods of development can lead to enduring changes in fetal and offspring neurodevelopment and behaviour. While there is emerging evidence from human studies to suggest a link between maternal negative mood states in pregnancy and various cognitive, behavioural, and emotional disturbances in offspring, it is not yet clear whether the programming mechanism demonstrated in animal studies also applies to humans. Few studies have directly assessed HPA axis function in the infants of prenatally stressed women. Research in this area has been constrained by a number of measurement challenges unique to the assessment of cortisol in infants. This paper discusses these challenges with a view to stimulating further research in the area. PMID- 17123742 TI - S-Citalopram in neuroendocrine challenge-tests: serotonergic responsivity in healthy male and female human participants. AB - The aim of the present study was to assess the usefulness of the selective serotonin-reuptake-inhibitor S-Citalopram as a serotonergic challenge probe in 24 healthy male and 24 healthy female participants. The participants received a single oral dose of 10 and 20mg of S-Citalopram in a placebo-controlled double blind crossover design. In female subjects phases of the menstrual cycle were controlled. Changes in concentrations of cortisol in saliva were used to indicate serotonergic reactivity. S-Citalopram induced a reliable dose dependent rise in cortisol concentrations. Results reveal a clear dose-response relationship in both sexes. However, in contrast to the 10mg condition the dosage of 20mg led to significantly higher cortisol levels in females, whereas no differences could be observed with respect to different phases of the menstrual cycle (follicular vs. luteal). Adverse side effects were reported only after 20mg. The results clearly indicate that the dosage of 10mg should be preferred when challenging males and females. Results will be discussed with regard to the further use of S-Citalopram in neuroendocrine challenge tests. PMID- 17123744 TI - Ethical conflicts in human embryonic stem cell research - a war between the "selfish genes"? PMID- 17123743 TI - A novel human detoxification system based on nanoscale bioengineering and magnetic separation techniques. AB - We describe the conceptual approach, theoretical background and preliminary experimental data of a proposed platform technology for specific and rapid decorporation of blood-borne toxins from humans. The technology is designed for future emergent in-field or in-hospital detoxification of large numbers of biohazard-exposed victims; for example, after radiological attacks. The proposed systems is based on nanoscale technology employing biocompatible, superparamagnetic nanospheres, which are functionalized with target-specific antitoxin receptors, and freely circulate within the human blood stream after simple intravenous injection. Sequestration of the blood-borne toxins onto the nanosphere receptors generates circulating nanosphere-toxin complexes within a short time interval; mathematical modeling indicates prevailing of unbound nanosphere receptors over target toxin concentrations at most therapeutic injection dosages. After a toxin-specific time interval nanosphere-toxin complexes are generated within the blood stream and, after simple arterial or venous access, the blood is subsequently circulated via a small catheter through a portable high gradient magnetic separator device. In this device, the magnetic toxin complexes are retained by a high gradient magnetic field and the detoxified blood is then returned back to the blood circulation (extracorporeal circulation). Our preliminary in vitro experiments demonstrate >95% first pass capture efficiency of magnetic spheres within a prototype high gradient magnetic separation device. Further, based on the synthesis of novel hydrophobic magnetite nanophases with high magnetization ( approximately 55 emu/g), the first biodegradable magnetic nanospheres at a size range of approximately 280 nm and functionalized with PEG-maleimide surface groups for specific antibody attachment are described here. In future applications, we envision this technology to be suitable for emergent, in-field usage for acutely biohazard exposed victims as both the injectable toxin-binding magnetic spheres and the separator device are made to be portable, light-weight, zero-power, and self- or helper-employed. Details of the technology are presented and the state-of-knowledge and research is discussed. PMID- 17123745 TI - Identification of genes regulated by transcription factor KLF7 in differentiating olfactory sensory neurons. AB - Gene targeting in mice has recently demonstrated that transcription factor KLF7 plays a critical role in neurite outgrowth and neuronal survival. Here we extended this genetic evidence by establishing the transcriptional profile of differentiating olfactory sensory neurons (OSNs) in Klf7(-/-) mice, and by identifying relevant genes that are directly regulated by KLF7. Functional clustering of DNA microarray data revealed that loss of KLF7 affects primarily the activity of genes involved in OSN differentiation, axonal growth, cytoskeletal dynamics, cell adhesion and synaptogenesis. Cell transfection experiments, on the other hand, demonstrated that the promoters of the genes encoding the OSN-specific OMP and the adhesion molecule L1 are both activated by KLF7 binding to CACCC motifs. Collectively, these results advance knowledge of transcriptional regulation of olfactory neurogenesis and KLF7 action. PMID- 17123746 TI - Prevalence of the initiator over the TATA box in human and yeast genes and identification of DNA motifs enriched in human TATA-less core promoters. AB - The core promoter of eukaryotic genes is the minimal DNA region that recruits the basal transcription machinery to direct efficient and accurate transcription initiation. The fraction of human and yeast genes that contain specific core promoter elements such as the TATA box and the initiator (INR) remains unclear and core promoter motifs specific for TATA-less genes remain to be identified. Here, we present genome-scale computational analyses indicating that approximately 76% of human core promoters lack TATA-like elements, have a high GC content, and are enriched in Sp1-binding sites. We further identify two motifs - M3 (SCGGAAGY) and M22 (TGCGCANK) - that occur preferentially in human TATA-less core promoters. About 24% of human genes have a TATA-like element and their promoters are generally AT-rich; however, only approximately 10% of these TATA containing promoters have the canonical TATA box (TATAWAWR). In contrast, approximately 46% of human core promoters contain the consensus INR (YYANWYY) and approximately 30% are INR-containing TATA-less genes. Significantly, approximately 46% of human promoters lack both TATA-like and consensus INR elements. Surprisingly, mammalian-type INR sequences are present - and tend to cluster - in the transcription start site (TSS) region of approximately 40% of yeast core promoters and the frequency of specific core promoter types appears to be conserved in yeast and human genomes. Gene Ontology analyses reveal that TATA less genes in humans, as in yeast, are frequently involved in basic "housekeeping" processes, while TATA-containing genes are more often highly regulated, such as by biotic or stress stimuli. These results reveal unexpected similarities in the occurrence of specific core promoter types and in their associated biological processes in yeast and humans and point to novel vertebrate specific DNA motifs that might play a selective role in TATA-independent transcription. PMID- 17123747 TI - Xenobiotic- and vitamin D-responsive induction of the steroid/bile acid sulfotransferase Sult2A1 in young and old mice: the role of a gene enhancer in the liver chromatin. AB - The xenobiotic-activated nuclear receptors PXR (pregnane X receptor) and CAR (constitutive androstane receptor) and the vitamin D(3)-activated nuclear receptor VDR regulate steroid and xenobiotic metabolism by inducing the phase I cytochrome P450 monooxygenases, phase II conjugating transferases, and the phase III transporters, which mediate the efflux of water-soluble lipid metabolites from cells. Metabolic stress due to the deviant expression of steroid- and xenobiotic-metabolizing enzymes is known to have severe health consequences including accelerated aging, and increased expression of these enzymes is associated with extended longevity [Gachon, F, Olela, FF, Schaad, O, Descombes, P and Schibler, U, 2006. The circadian PAR-domain basic leucine zipper transcription factors DBP, TEF, and HLF modulate basal and inducible xenobiotic detoxification. 4, 25-36.; McElwee, JJ, Schuster, E, Blanc, E, Thomas, JH and Gems, D, 2004. Shared Transcriptional Signature in Caenorhabditis elegans Dauer Larvae and Long-lived daf-2 Mutants Implicates Detoxification System in Longevity Assurance. J. Biol. Chem., 279, 44533-43.]. Information on the similarities and dissimilarities in drug metabolism between the young and old, as may be uncovered by studying aging regulation of the genes relevant to steroid and xenobiotic metabolism, is likely to have clinical significance. In this report, we examined the VDR- and PXR-mediated gene induction of the phase II sulfotransferase Sult2A1 in the livers of 4-month- and 20-month-old mice. Sult2A1 converts bile acids, steroids and a number of drugs to the corresponding sulfated metabolites, which are readily eliminated from the body due to increased water solubility. In RT-PCR assay, aging did not change the induction of Sult2A1 mRNAs by the hormonally active vitamin D(3) and the catatoxic synthetic steroid PCN (pregnenolone-16alpha carbonitrile). Chromatin immunoprecipitation (ChIP) from liver nuclei showed that aging had no effect on the activity of an IR0 enhancer in the Sult2A1 chromatin to recruit VDR, RXR-alpha (retinoid X receptor) and PXR in mice injected with D(3) or PCN. Thus, mice in late life are as competent as those in early life in responding to the hormonal and xenobiotic signaling for Sult2A1 induction. This is the first report describing the role of aging in the functional response of an enhancer in the liver chromatin to the nuclear receptor-dependent signaling. PMID- 17123748 TI - The mitochondrial genome of Priapulus caudatus Lamarck (Priapulida: Priapulidae). AB - We sequenced and annotated the complete mitochondrial (mt) genome of the priapulid Priapulus caudatus in order to provide a source of phylogenetic characters including an assessment of gene order arrangement. The genome was 14,919 bp in its entirety with few, short non-coding regions. A number of protein coding and tRNA genes overlapped, making the genome relatively compact. The gene order was: cox1, cox2, trnK, trnD, atp8, atp6, cox3, trnG, nad3, trnA, trnR, trnN, rrnS, trnV, rrnL, trnL(yaa), trnL(nag), nad1, -trnS(nga), -cob, -nad6, trnP, -trnT, nad4L, nad4, trnH, nad5, trnF, -trnE, -trnS(nct), trnI, -trnQ, trnM, nad2, trnW, -trnC, -trnY; where '-' indicates genes transcribed on the opposite strand. The gene order, although unique amongst Metazoa, shared the greatest number of gene boundaries and the longest contiguous fragments with the chelicerate Limulus polyphemus. The mt genomes of these taxa differed only by a single inversion of 18 contiguous genes bounded by rrnS and trnS(nct). Other arthropods and nematodes shared fewer gene boundaries but considerably more than the most similar non-ecdysozoan. PMID- 17123749 TI - NgUNC-119, Naegleria homologue of UNC-119, localizes to the flagellar rootlet. AB - The UNC-119 family of proteins is ubiquitous in animals. The expression of UNC 119 is prominent in neural tissues including photoreceptor cells. Homologues of UNC-119 are also found in ciliated (or flagellated) single-celled organisms; however, the cellular distribution of this protein in protists is unknown. We cloned and characterized a homologue of unc-119 from the ameboflagellate Naegleria gruberi (Ngunc-119) and identified the cellular distribution of the protein. The Ngunc-119 open reading frame contained 570 nucleotides encoding a protein of 189 amino acids with a predicted molecular weight of 22.1 kDa, which is similar to that of Paramecium UNC-119 and Trypanosoma UNC-119. These three proteins are 46-48% identical in their amino acid sequences. The smaller NgUNC 119 corresponds to the conserved C-terminal 3/4 of the UNC-119 from multi cellular organisms. The amino acid sequence of NgUNC-119 is 43-50% identical to that of the conserved C-terminal regions. NgUNC-119 was not found in growing amoebae but accumulated rapidly after the initiation of differentiation into flagellates. Indirect immunofluorescence staining of differentiating N. gruberi showed that NgUNC-119 begins to concentrate at a spot near the nucleus of differentiating cells and then elongates into a filamentous structure. Purification and indirect immunofluorescence staining of the Naegleria flagellar rootlet suggested that NgUNC-119 is a component of the flagellar rootlet. PMID- 17123750 TI - Ehrlichia ruminantium variants which do not cause heartwater found in South Africa. AB - In 1994 a batch of apparently healthy goats was selected for intended export to the USA from a heartwater-free and vector tick-free region of South Africa. The animals were tested serologically for heartwater, using either or both an IFA and an ELISA test, and 52% were found to be serologically positive. A PCR assay based on Ehrlichia ruminantium 16S gene sequences gave positive results for 54% of the animals, suggesting that apparently non-pathogenic E. ruminantium variants existed in this heartwater-free area. To identify and characterise the agents responsible for the positive serological and PCR results, ticks and animal blood samples were collected from two of the three farms involved in the original survey during two successive seasons of expected peak tick activity. Ticks were kept alive for a minimum of 3 weeks to allow digestion of any blood meal before being processed. Over the two seasons, 28% of the livestock and 15% of the ticks sampled were found to be carrying E. ruminantium. E. ruminantium 16S and pCS20 sequences were detected in all of the four tick species collected from the livestock (Rhipicephalus evertsi evertsi, Rhipicephalus evertsi mimeticus, Hyalomma truncatum, Hyalomma marginatum rufipes), suggesting that some of the species may act as vectors. Animals generally carried multiple E. ruminantium 16S genotypes, whereas ticks rarely carried more than one. Infection levels in both animals and ticks were too low to generate a marked response when a blood stabilate was sub-passaged in a clean sheep, preventing the subsequent establishment of any of the organisms in culture. PMID- 17123751 TI - Efficacy of spheroplastic and cell-wall competent vaccines for Mycobacterium avium subsp. paratuberculosis in experimentally-challenged baby goats. AB - A Mycobacterium avium subspecies paratuberculosis (MAP) vaccine that reduced the incidence of clinical disease or reduced fecal shedding of MAP would aid control of Johne's disease (JD). The objective of the present study was to evaluate the efficacy of four MAP vaccine combinations, including cell-wall competent (CWC) alum adjuvant, CWC-QS21 adjuvant, cell-wall deficient (CWD) alum adjuvant and CWD QS21 adjuvant vaccines. Eighty baby goats were vaccinated at 1 and 4 weeks of age with one of these vaccines or a sham control vaccine consisting of alum adjuvant. Kids were challenged orally with approximately 6.0x10(9) organisms in four divided doses of 1.5x10(9) organisms using a goat isolate of MAP. Vaccinated challenged and challenged control groups had 10 and 6 kids per group, respectively. Half of the kids within each group were necropsied at either 6 or 9 months post-challenge. Gross and microscopic lesions and relative number of acid fast bacilli were evaluated and scored at necropsy. Results indicated all challenged kids had some lesions compatible with JD suggesting none of the vaccines prevented infection. Three vaccines (CWC-alum, CWC-QS21 and CWD-QS21) reduced lesion scores by 46-51% at 9 months. CWD-alum vaccine resulted in a more severe (+33.5%) lesion score than sham-vaccinated challenged control. Lesion scores were greater at 9 months than at 6 months post-challenge in the sham vaccinated challenged group and CWD-alum vaccinated group, while lesion scores were generally stable with remaining vaccines. Mean fecal CFU/g were significantly different across time from challenge to 9-month necropsy (p=0.043) and the CWC-QS21 vaccine group had a marked reduction in fecal CFU/g at all time points post-challenge. A reduction in MAP CFU/g was also detected in necropsy tissues from kids given the CWC-alum, CWC-QS21 and CWD-QS21 vaccines, and increased CFU/g were detected in tissues from kids given the CWD-alum vaccine. Immunological tests evaluated included, humoral response evaluation by AGID, ELISA and Western blot, and cell mediated response by comparative PPD skin testing (M. avium, Old Johnin, M. bovis and Lot 2 Johnin PPD's), and production of MAP induced gamma-interferon. Vaccination also resulted in false-positive PPD skin test reactions for M. avium PPD, Old Johnin PPD and gamma-interferon tests. When a 2-mm cutoff above normal skin thickness was used to define positive skin test reactions, false-positive reactions for M. bovis were detected in only 2 of 32 kids given a vaccine with QS21 adjuvant. PMID- 17123752 TI - Interactions of highly and low virulent Mycoplasma hyopneumoniae isolates with the respiratory tract of pigs. AB - Mycoplasma hyopneumoniae is the etiological agent of swine enzootic pneumonia, a chronic nonfatal disease affecting pigs of all ages. To obtain better insight in the mechanisms responsible for differences in virulence between highly and low virulent M. hyopneumoniae isolates, 23 caesarean-derived, colostrum-deprived piglets were randomly assigned to three groups. Groups 1 and 2 consisted of nine animals each, which were intratracheally inoculated at 1 week of age with a highly or a low virulent isolate of M. hyopneumoniae, respectively. The remaining five animals were inoculated with sterile culture medium. Animals were euthanized at 5, 10, 15 and 28 days post-inoculation (DPI). Animals inoculated with the highly virulent isolate had more neutrophils in BAL fluid at 10, 15 and 28DPI compared to the other groups. At 10 and 15DPI, animals in the highly virulent group had significantly higher concentrations of TNF-alpha in BAL fluid. IL-1beta concentration in this group was higher at 5 and 28DPI compared to the other groups. From 10DPI onwards, significantly higher titres of M. hyopneumoniae were detected in the BAL fluid of animals inoculated with the highly virulent isolate compared to animals inoculated with the low virulent isolate. Additionally, the in vitro generation time of the highly virulent M. hyopneumoniae isolate was significantly shorter than that of the low virulent isolate. The present study indicates that the difference in pathogenicity between the highly and low virulent isolates is associated with a faster in vitro growth, a higher capacity to multiply in the lungs and the induction of a more severe inflammation process by the highly virulent isolate. PMID- 17123753 TI - Investigation of the immunogenicity of diclofenac and diclofenac metabolites. AB - Oral administration of the non-steroidal anti-inflammatory drug diclofenac (DCF) is associated with a high incidence of adverse drug reactions, some of which are thought to be mediated by the immune system. It has been proposed that metabolic activation of DCF and covalent binding to protein generates an antigenic determinant that stimulates immune cells; however, the nature of the metabolite remains ill-defined. The aim of this study was to synthesize and evaluate the antigenic potential of DCF metabolites in the mouse. DCF and DCF metabolites were administered via subcutaneous injection over a 5-day period to BALB/C strain mice to induce immune activation. Proliferation was measured by the addition of [(3)H] thymidine to ex vivo isolated draining auricular lymph node cells. Results were compared with those provoked by exposure to 2,4-dinitrochlorobenzene. Lymph node activation was observed following treatment with 2,4-dinitrochlorobenzene, 5 hydroxy DCF quinoneimine and 4'-hydroxy DCF quinoneimine, but not DCF acyl glucuronide or DCF itself. Interestingly, lymph node cells from 5-hydroxy DCF treated mice were also found to proliferate, when compared with cells from vehicle-treated mice, while 4'-hydroxy DCF did not stimulate lymph node cell activation. The reactivity of 5-hydroxy DCF quinoneimine was confirmed by synthesis and characterization of an N-acetyl cysteine adduct. These data show that formation of 5-hydroxy DCF and subsequent autoxidation provides an antigenic determinant for immune cell activation in the mouse. PMID- 17123754 TI - Differential effects of cadmium on the gene expression of seven-transmembrane spanning receptors and GAPDH in the rat testis. AB - Cadmium (Cd) is a widely spread toxicant with endocrine disrupting properties. Under experimental conditions it suppresses sex steroid synthesis in the male as well as the female. Testicular steroidogenesis is primarily regulated by gonadotropins, but is also influenced by catecholamines. We have previously shown that Cd exposure affects rat testosterone synthesis by down-regulating luteinizing hormone (LH) receptor mRNA expression. In this study, rats were given 10 micromol/kg Cd subcutaneously and sacrificed 0.48-144 h later. We investigated the effects of Cd on testicular gene expression of two adrenergic receptors. In addition, mRNA levels of the androgen-regulated house keeping gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were measured. In contrast to the suppressive influence on LH receptor expression Cd lacked effect on the expression of alpha(1A)- and beta(2)-adrenergic receptors. GAPDH gene expression, on the other hand, was up-regulated 1.6-fold after exposure to 10 micromol/kg Cd. These data suggest that the influence of Cd on testicular gene expression involves a specific effect on the LH receptor and not a general effect on seven transmembrane-spanning receptors. Also, data indicate that the increased expression of GAPDH may be secondary to Cd-induced testosterone deprivation, suggesting future studies of androgen-regulated genes in the toxicity of Cd. PMID- 17123755 TI - Horn growth related to testosterone secretion in two wild Mediterranean ruminant species: the Spanish ibex (Capra pyrenaica hispanica) and European mouflon (Ovis orientalis musimon). AB - Seasonal variations in the horn development and testicular activity of the Spanish ibex (Capra pyrenaica hispanica) (n=6) and European mouflon (Ovis orientalis musimon) (n=5) were monitored to determine the role of increasing testosterone concentration on the arrest of horn growth during the rutting season. Marked seasonal variations in the rate of horn growth (P<0.01) and testicular activity (P<0.001) were seen in both species, although the magnitude and timing of these changes were different (P<0.01). Horn growth rate was inversely correlated to seasonal levels in testosterone plasma concentration in both species (ibex: R=-0.45, P<0.01; mouflon R=-0.51, P<0.01). In the mouflon, the increase in plasma testosterone concentration recorded in September (P<0.05 compared with the lowest concentration) coincided with a significant reduction in horn growth (P<0.05). In the ibex, the increase in plasma testosterone concentration in October (P<0.05 compared with the lowest concentration) was associated with a significant arrest of horn growth in November (P<0.05). These results appear to support the hypothesis that high peripheral plasma levels of testosterone are linked with the seasonal arrest of horn growth during the rutting period. PMID- 17123756 TI - True vaginal prolapse in a bitch. AB - Frequently, vaginal fold prolapse is the protrusion of edematous vaginal tissue into and through the opening of the vulva occurring during proestrus and estrus stages of the sexual cycle. True vaginal prolapse may occur near parturition, as the concentration of serum progesterone declines and the concentration of serum oestrogen increases. In the bitch, this type of true vaginal prolapse is a very rare condition. This short communication describes a 5-year-old female, cross breed dog in moderate condition, weighing 33 kg, with distocia and true vaginal prolapse. Abdominal palpation and transabdominal ultrasonography revealed live and dead foetuses in the uterine horns. One dead and four live fetuses were removed from uterus by cesarean section. The ovariohysterectomy was performed after repositioning the vaginal wall with a combination of traction from within the abdomen and external manipulation through the vulva. Re-occurrence of a vaginal prolapse was not observed and the bitch recovered completely after the surgical therapy. Compared to other vaginal disorders, vaginal prolapse is an uncommon condition in the bitch. In the present case, extreme tenesmus arising from distocia may have predisposed to the vaginal prolapse. The cause of dystocia was probably the disposition of the first foetus. We concluded that the vaginal prolapse was the result of dystocia in the present case. PMID- 17123757 TI - Physical stability and aerosol properties of liposomes delivered using an air-jet nebulizer and a novel micropump device with large mesh apertures. AB - The aerosol properties of liposomes and their physical stability to aerosolization were evaluated using an air-jet nebulizer (Pari LC Plus) and a customized large aperture vibrating-mesh nebulizer (Aeroneb Pro-8microm). Soya phosphatidylcholine: cholesterol (1:1 mole ratio) multilamellar liposomes (MLVs) entrapping salbutamol sulfate were nebulized directly, or after being reduced in size by extrusion through 1 or 0.4microm polycarbonate membrane filters. MLVs were very unstable to jet nebulization and stability was not markedly enhanced when vesicles were extruded before nebulization, such that drug losses from delivered liposomes using the Pari nebulizer were up to 88% (i.e. only 12% retained in liposomes). The Aeroneb Pro-8microm nebulizer was less disruptive to liposomes, completed nebulization in a much shorter time, and produced greater mass output rate than the Pari nebulizer. However, aerosol droplets were larger, total drug and mass outputs were lower and aerosolization performance was dependent on formulation. Vibrating-mesh nebulization was less disruptive to liposomes extruded through the 1microm membranes compared with the non-extruded MLVs, so that the retained entrapment of the drug in the nebulized vesicles was 56% and 37%, respectively. However, extrusion of liposomes to 0.4microm resulted in reduced stability of liposomes to vibrating-mesh nebulization (retained entrapment=41%) which was attributed to the reduced liposome lamellarity and subsequent reduced resistance to nebulization-induced shearing. This study has shown that vibrating-mesh nebulization using the customized large aperture mesh nebulizer (Aeroneb Pro-8microm) had a less disruptive effect on liposomes and produced a higher output rate compared with the Pari LC Plus air-jet nebulizer. On the other hand, the air-jet nebulizer produced higher total mass and drug outputs and smaller aerosol droplets. PMID- 17123758 TI - Mild noise-induced hearing loss at young age affects temporal modulation transfer functions in adult cat primary auditory cortex. AB - Kittens were exposed for 2h to a 1/3rd octave band of noise centered at 5kHz and at 120dB SPL. After the exposure, they were kept in a quiet room for at least 4 weeks, and until they were mature. The noise-exposed cats showed on average 16.5dB higher ABR thresholds and 13.2dB higher thresholds at the characteristic frequency (CF) than the control cats for frequencies between 4 and 16kHz. The frequency-tuning curve bandwidth at 20dB above threshold was significantly increased compared to controls in the CF region of the hearing loss. In noise exposed cats, temporal modulation-transfer functions (tMTFs) to amplitude modulated (AM) noise, but not to periodic click trains, showed a marked increase for modulation frequencies (MFs) below 6Hz. The vectorstrength in noise-exposed cats increased for all modulation frequencies below 32Hz for neurons with a CF in the range of the hearing loss. The tMTFs for AMnoise in the noise-exposed group were less band-pass compared to the controls, and in that sense the mild hearing loss could be considered as effectively reducing the central activation in the same way as a reduced sound pressure level. Effects of reduced central inhibition are visible in the broadening of frequency-tuning curves, and in the increased limiting rates for AMnoise. PMID- 17123759 TI - Bioassay-guided isolation of anti-inflammatory and antinociceptive glycoterpenoids from the flowers of Verbascum lasianthum Boiss. ex Bentham. AB - Infusions of Verbascumlasianthum flowers have been used for hemorrhoids in Turkish folk medicine. In order to evaluate the scientific basis for this practice, in vivo anti-inflammatory and antinociceptive activities of Verbascum lasianthum Boiss. ex Bentham flowers were investigated. A methanolic extract of the flowers was shown to possess significant inhibitory activity in the carrageenan-induced hind paw edema model and in p-benzoquinone-induced writhings in mice. Through bioassay-guided fractionation and isolation procedures eight compounds, 6-O-(4'''-O-trans-p-coumaroyl)-alpha-L-rhamnopyranosylaucubin (1), 6-O (4'''-O-trans-p-methoxycinnamoyl)-alpha-L-rhamnopyranosylaucubin (2), sinuatol (3), aucubin (4), geniposidic acid (5), catalpol (6), ajugol (7) and ilwensisaponin A (8) were isolated and their structures were elucidated by spectral techniques. An iridoid glucoside, aucubin (4) and a triterpenoid saponin, ilwensisaponin A (8) were found to possess significant antinociceptive and anti-inflammatory activities, per os without inducing any apparent acute toxicity or gastric damage. Results of the present study support the continued and expanded utilization of plants employed in Turkish folk medicine. PMID- 17123760 TI - Glabrol, an acyl-coenzyme A: cholesterol acyltransferase inhibitor from licorice roots. AB - Acyl-coenzyme A: cholesterol acyltransferase (ACAT) esterifies free cholesterol in the liver and the intestine. It has relations with production of lipoproteins and accumulation of cholesteryl esters of the atheroma. Therefore, ACAT inhibitors may act as antihypercholesterolemic and antiatherosclerotic agents. One isoprenyl flavonoid was isolated from ethanol extract of licorice roots. On the basis of spectral evidences, the compound was identified as glabrol (1). Compound 1 inhibited rat liver microsomal ACAT activity with an IC(50) value of 24.6 microM and decreased cholesteryl ester formation with an IC(50) value of 26.0 microM in HepG2 cells. In addition, 1 showed a non-competitive type of inhibition against ACAT. PMID- 17123761 TI - Protective effects of keishibukuryogan on the kidney of spontaneously diabetic WBN/Kob rats. AB - Keishibukuryogan, one of the traditional herbal formulations, is used clinically to improve blood circulation. It consists of the following five crude drugs: Cinnamomi Cortex, Poria, Moutan Cortex, Persicae Semen and Paeoniae Radix. In this study, the effects of keishibukuryogan against renal damage in spontaneously diabetic WBN/Kob rats were examined. Oral administration of keishibukuryogan significantly attenuated urinary protein excretion and serum creatinine levels. It did not affect body weight loss and blood glucose levels, but it suppressed renal and hepatic weights of WBN/Kob rats. Keishibukuryogan also reduced fibronectin and transforming growth factor beta(1) (TGF-beta(1)) protein expression in the renal cortex. Furthermore, lipid peroxidation levels in both kidney and liver were significantly lower than those of untreated control WBN/Kob rats. Urinary excretion of 8-hydroxy-deoxyguanosine was suppressed by keishibukuryogan treatment. These results suggest that keishibukuryogan reduces oxidative stress by hyperglycemia, and that it protects renal function and suppresses fibronectin deposition induced by TGF-beta(1) production in WBN/Kob rats. PMID- 17123762 TI - Imaging of peripheral arteries by 16-row multidetector computed tomography angiography: a feasible tool? AB - OBJECTIVE: To evaluate the efficacy of multidetector (16-row) computed tomography (MDCT) in imaging the upper and lower limb arterial tree in trauma and peripheral arterial occlusive disease (PAOD). METHODS: Thirty-three patients underwent MDCT angiography (MDCTA) of the upper or the lower limb on 16-row MDCT scanner between November, 2004 and July, 2005. The findings were compared with the surgical outcome in cases with trauma and suspected arterial injuries or color Doppler correlation was obtained for patients of PAOD. RESULTS: MDCTA allowed a comprehensive diagnostic work-up in all trauma cases with suspected arterial injuries. In the 23 cases of PAOD, MDCT adequately demonstrated the presence of stenosis or occlusion, its degree and extent, the presence of collaterals and plaques. CONCLUSION: Our experience of CT angiography (CTA) with 16-row MDCT scanner has clearly demonstrated its efficacy as a promising, new, fast, accurate, safe and non-invasive imaging modality of choice in cases of trauma with suspected arterial injuries and as a useful screening modality in cases of PAOD for diagnosis and for grading. PMID- 17123763 TI - Multidetector CT and MRI findings in periportal space pathologies. AB - Periportal region is an anatomic space around portal vein comprising hepatic artery, bile duct, nerves, lymphatics and a potential space. Periportal pathologies may involve any of these structures diffusely or focally with characteristic radiologic findings. Radiologic findings can be helpful in differential diagnosis of pathologies of periportal structures including periportal cavernomatous transformation, hepatic artery aneurysm, biliary diseases, neurofibromatosis, lymphoma, langerhans' cell histiocytosis, periportal fatty infiltration and other causes of periportal halo in adult and pediatric patients. Lobar/segmental intrahepatic involvement can be seen in neurofibromatosis, cavernomatous transformation, fatty infiltration and periportal edema. In this review, we discuss CT and MRI findings of periportal pathologies which can be in the form of diffuse or segmental/lobar involvement. PMID- 17123764 TI - Budd-Chiari syndrome: a review of imaging findings. AB - Budd-Chiari syndrome is an uncommon, often fatal disorder resulting from an obstructed hepatic venous outflow tract. The obstructive lesion is situated in the main hepatic veins, in the inferior vena cava or in both. The nature, location and extension of the obstruction can be displayed on diagnostic imaging techniques. In addition to this direct evidence, the indirect findings of venous obstruction such as the presence of intra- and extrahepatic collateral veins, when combined with the altered morphology and enhancement pattern of the liver enables one to arrive at a confident diagnosis. In patients with suspected Budd Chiari syndrome, gray-scale sonography with complementary support of color and pulsed Doppler examinations is the first step in approaching the diagnosis. It is followed by a contrast-enhanced cross-sectional technique, preferrentially by MR angiography. The patients with a high clinical suspicion of Budd-Chiari syndrome may undergo hepatic venography or venacavography directly so that a potential of recanalization (e.g. percutaneous transluminal angioplasty with or without stent placement or TIPS) of the obstructed segment under the guidance of these techniques would not be delayed. PMID- 17123765 TI - Popliteal artery volume flow measurement: a new and reliable predictor of early patency after infrainguinal balloon angioplasty and subintimal dissection. AB - OBJECTIVE: We have investigated whether popliteal artery volume flow (PAVF) measured immediately after balloon angioplasties of the superficial femoral artery-popliteal segments (SFA/POP) was predictive of early (30 days) and mid term (6 months) arterial thrombosis. METHODS: During the last 24 months, 203 patients (56% men) with a mean age of 73 +/- 9 years had 268 duplex-guided balloon angioplasties of the SFA/POP. Critical ischemia was the indication in 36%. Group I included 176 (66%) with stenoses, and group II had 92 (34%) with occlusions. All patients had completion duplex examinations that included three measurements of PAVF of below-the-knee popliteal artery. RESULTS: Early (30 days) thrombosis of the treated femoropopliteal arterial segment developed in 10 patients (3.7%), three in group I (1.7%) and seven in group II (7.6%; P < .04). All 10 cases of early thrombosis were in patients with TransAtlantic Inter Society Consensus (TASC) class C (6/185, 3.2%) and D (4/26, 15%) lesions. Moreover, the 19% incidence (n = 4) of early thrombosis in patients with PAVF <100 mL/min (mean, 73 +/- 24 mL/min; range, 20 to 99 mL/min) was higher compared with the 2.4% rate for patients with higher flows (mean, 176 +/- 60 mL/min; range, 100 to 450 mL/min; P < .01). At 6 months of follow-up, femoropopliteal occlusions had developed in nine more patients, and it became apparent that low PAVF measurements were still predictive of thrombosis (29%) when compared with higher PAVF cases (6%; P < .002). Log-rank comparison of survival curves for cumulative primary stenosis-free patency in group I and group II demonstrated a statistically significant difference (P < .02). PAVF <100 mL/min and TASC classification were significant predictors of early (30 days) and mid-term (6 months) arterial thrombosis after femoropopliteal angioplasties. PAVF was the most powerful predictor of arterial thrombosis. The respective 6-month and 12 month limb salvage rates were 98% and 94% for patients with claudication and 88% and 85% for those with limb-threatening ischemia (P < .0001). CONCLUSIONS: Our results demonstrate that low PAVF is the most powerful predictor of early (30 days) and mid-term (6 months) arterial thrombosis after femoropopliteal interventions. In the presence of a low postprocedure PAVF (<100 mL/min), one may consider not reversing the heparin or using intermittent calf compression, or both, to augment the arterial flow. PMID- 17123767 TI - Does the timing of reoperation influence the risk of graft infection? AB - OBJECTIVE: This study compared the incidence and characteristics of graft infection in patients who underwent early vs late revisional surgery of lower extremity arterial bypass grafts. METHODS: Between 1992 and July 2005, 500 revisional procedures were performed on 198 lower extremity bypass grafts. Patients whose revisions were performed <30 days after the primary bypass were in the early revision (ER) group (n = 99), and those done >30 days after bypass were in the late revision (LR) group (n = 99). Infection was defined as cellulitis with graft exposure or purulence in continuity with a graft that required antibiotics and operation for infection control. Mean follow-up was 60 months (range, 2 to 60 months). Groups were compared using Student's t test. RESULTS: The ER group included 66 autogenous and 33 prosthetic grafts. The LR group consisted of 53 autogenous and 46 prosthetic grafts. Of the 500 revisional procedures performed, 17 graft infections occurred (3.4%). Twelve (70.6%) were prosthetic grafts and five (29.4%) were autogenous grafts (P = .004). Defining the infection rate per graft rather than per revisional procedure, the ER group had a significantly higher graft infection rate at 11% (11/99) compared with 6.1% in the LR group (6/99; P = .012). The risk of infection for prosthetic grafts was significantly higher within the ER group at 27.3% (9/33) compared with autogenous grafts at 3.1% (2/66; P = .0001). Infection developed in three vein grafts and three prosthetic grafts in the LR group (P = NS). For prosthetic graft revisions only, infection risk was 27.3% (9/33) in the ER group and 6.5% (3/46) in the LR group (P = .005). The most common cultured pathogen was methicillin resistant Staphylococcus aureus (ER, 6/11 vs LR, 3/6; P = NS). Within the ER group, the prevalence of Pseudomonas aeruginosa was significantly higher at 27.3% (3/11) compared with 0% (0/6) in the LR group (P = .04). CONCLUSIONS: Early revision of lower extremity arterial bypass grafts has a significantly higher risk of graft infection compared with revision >1 month after surgery. Infection will develop in approximately 25% (9/33) of prosthetic grafts that are reoperated on early. If feasible, reoperation should be delayed >1 month for prosthetic grafts needing revision. Endovascular or extra-anatomic interventions should be considered if early revision is mandated in this group. PMID- 17123769 TI - Elevated C-reactive protein levels are associated with postoperative events in patients undergoing lower extremity vein bypass surgery. AB - OBJECTIVES: Inflammatory markers such as high-sensitivity C-reactive protein (hsCRP) are associated with an increased risk of cardiovascular events and with the severity of peripheral arterial disease. The effects of inflammation on the development of vein graft disease remain speculative. We hypothesized that high levels of inflammatory markers would identify patients at increased risk for adverse events (graft failure, major cardiovascular events) after lower extremity bypass surgery. METHODS: Patients (n = 91) scheduled to undergo lower extremity bypass using autogenous vein were enrolled into a prospective study at two institutions. Exclusion criteria included the presence of major infection. A baseline plasma sample was obtained on the morning of lower extremity bypass. Biomarkers for inflammation included hsCRP, fibrinogen, and serum amyloid A (SAA). Values between patients with and without critical limb ischemia were compared. Proportions of events among dichotomized populations (upper limit of normal of each laboratory assay) were compared by log-rank test. RESULTS: Of the patients undergoing lower extremity bypass, 69% were men, 53% were diabetic, 81% were smokers, and their mean ankle-brachial index was 0.51 +/- 0.19. The indication for lower extremity bypass was critical limb ischemia in 55%. There were no perioperative deaths and two early graft occlusions. During a mean follow up of 342 days (range, 36-694 days) there were four deaths, 27 graft-related events, and 10 other cardiovascular events. No relationships were found between events and demographics, comorbidities, baseline ankle-brachial index, or statin use. High-sensitivity CRP (P = .005), fibrinogen (P < .001), and SAA (P = .0001) levels were associated with critical limb ischemia at presentation. Among patients with an elevated hsCRP (>5 mg/L) immediately before surgery, major postoperative vascular events occurred in 60% (21/35), compared with a 32% (18/56) rate in those with a baseline CRP <5 mg/L (P = .004, log-rank test). On multivariable analysis, only elevated hsCRP correlated with adverse graft-related or cardiovascular events (P = .018). CONCLUSIONS: The inflammatory biomarkers of hsCRP, fibrinogen, and SAA correlate with peripheral arterial disease severity at presentation in patients undergoing lower extremity bypass. Patients with elevated hsCRP are at increased risk for postoperative vascular events, most of which are related to the vein graft. These findings suggest a potential relationship between inflammation and outcomes after lower extremity vein bypass surgery. PMID- 17123770 TI - Association of obesity and metabolic syndrome with the severity and outcome of intermittent claudication. AB - BACKGROUND: Obesity is recognized as an independent predictor of coronary artery disease; however, its importance in peripheral arterial disease is less clear. The aim of this study was to assess the association between obesity and the severity and outcome of intermittent claudication. METHODS: This study was a prospective cohort study based at a tertiary referral center. Sixty patients with intermittent claudication selected for conservative treatment were assessed for obesity and metabolic syndrome by using the International Diabetes Federation definition. Other risk factors, including diabetes, hypertension, smoking history, serum lipids, adipocytokines, and C-reactive protein, were measured by clinical and blood assessment. Obesity and metabolic syndrome were related to the severity of peripheral arterial disease, defined by ankle-brachial pressure index and graded treadmill measured maximum walking distance (MWD) and initial claudication distance, by using multiple linear regression analysis allowing for traditional atherosclerotic risk factors. Patients were followed up for 24 months, and combined outcome was reported in terms of death, cardiovascular events, or requirement for revascularization. The effect of obesity and metabolic syndrome on outcome was investigated by using Kaplan-Meier and Cox proportional hazard analysis. RESULTS: Obesity and serum adiponectin were independently associated with the severity of peripheral arterial disease measured by ankle brachial pressure index (P = .03 and .001), initial claudication distance (P = .009 and .03), and MWD (P = .001 and .04). Metabolic syndrome was independently associated only with MWD (P = .02). By 24 months, outcome events occurred in 37% +/- 7% and 43% +/- 9% of patients with metabolic syndrome or obesity, respectively, compared with 0% and 11% +/- 6% of those without these diagnoses. Waist circumference independently predicted the likelihood of outcome events (relative risk, 1.16; 95% confidence interval, 1.08-1.26; P < .001). CONCLUSIONS: These findings, if confirmed in other cohorts, suggest the importance of treating obesity in patients with intermittent claudication. Serum adiponectin concentrations may be an important guide to the efficacy of treatment in patients with intermittent claudication and obesity. PMID- 17123771 TI - Local recurrence in breast carcinoma patients with T(1-2) and 1-3 positive nodes: indications for radiotherapy. AB - AIMS: To investigate the relationship between local recurrence (LR) and distant recurrence (DR) and to determine a subgroup of patients who could benefit from radiotherapy among breast carcinoma patients with T(1-2) and N(1a). METHODS: Univariate and multivariate Cox regression analyses were carried out in the retrospective data of 326 eligible patients. RESULTS: Fourteen (4.3%) patients had LR and 46 (14.1%) patients suffered DR, in their follow-up periods. The multivariate time-dependent Cox model for DR showed that ratio of positive nodes (PN) (p=0.004; hazard ratio (HR), 1.05; 95% confidence interval (CI), 1.02-1.09) and LR (p=0.05; HR, dependent on time) were strongly associated with DR. In the multivariate Cox analysis for LR, age (35 years; p<0.0001; HR, 6.8; CI, 2.3-19.9), lymphatic vascular invasion (LVI) (yes vs no; p=0.03; HR, 3.3; CI, 1.2-9.8), and a ratio of PN (>15% vs 80% decrease in SHIP2 mRNA and 60-80% decrease in SHIP2 protein, which resulted in significant gene expression changes linked to cytoskeletal functions, including altered expression of adducin-alpha, pallidin, stathmin-like-2, and synaptojanin 2 binding protein. Insulin treatment of C2C12 muscle cells caused transcriptional changes associated with known signaling pathways. However, SHIP2 depletion had no discernible effect on insulin-regulated gene expression. Taken together, our results suggest that SHIP2 is involved in the regulation of cytoskeletal functions, but a large reduction of SHIP2 in C2C12 muscle cells is not sufficient to affect insulin-mediated gene expression. PMID- 17123778 TI - Induction of mammary gland ductal hyperplasias and carcinoma in situ following fetal bisphenol A exposure. AB - Exposure of the fetus to excess estrogen is believed to increase the risk of developing breast cancer during adult life. Fetal exposure to low doses of the xenoestrogen bisphenol A resulted in long-lasting effects in the mouse mammary gland that were manifested during adult life. It enhanced sensitivity to estradiol, decreased apoptosis, increased the number of progesterone receptor positive epithelial cells at puberty and increased lateral branching at 4 months of age. We now report that fetal exposure to 2.5, 25, 250 and 1000 microg bisphenol A/kg body weight/day induces the development of ductal hyperplasias and carcinoma in situ at postnatal day 50 and 95 in rats. These highly proliferative lesions have an increased number of estrogen receptor-alpha positive cells. Thus, fetal bisphenol A exposure is sufficient to induce the development of preneoplastic and neoplastic lesions in the mammary gland in the absence of any additional treatment aimed at increasing tumor development. PMID- 17123779 TI - Developmental estrogen exposures predispose to prostate carcinogenesis with aging. AB - Prostate morphogenesis occurs in utero in humans and during the perinatal period in rodents. While largely driven by androgens, there is compelling evidence for a permanent influence of estrogens on prostatic development. If estrogenic exposures are abnormally high during the critical developmental period, permanent alterations in prostate morphology and function are observed, a process referred to as developmental estrogenization. Using the neonatal rodent as an animal model, it has been shown that early exposure to high doses of estradiol results in an increased incidence of prostatic lesions with aging which include hyperplasia, inflammatory cell infiltration and prostatic intraepithelial neoplasia or PIN, believed to be the precursor lesion for prostatic adenocarcinoma. The present review summarizes research performed in our laboratory to characterize developmental estrogenization and identify the molecular pathways involved in mediating this response. Furthermore, recent studies performed with low-dose estradiol exposures during development as well as exposures to environmentally relevant doses of the endocrine disruptor bisphenol A show increased susceptibility to PIN lesions with aging following additional adult exposure to estradiol. Gene methylation analysis revealed a potential epigenetic basis for the estrogen imprinting of the prostate gland. Taken together, our results suggest that a full range of estrogenic exposures during the postnatal critical period - from environmentally relevant bisphenol A exposure to low-dose and pharmacologic estradiol exposures - results in an increased incidence and susceptibility to neoplastic transformation of the prostate gland in the aging male which may provide a fetal basis for this adult disease. PMID- 17123780 TI - DNA microarray for the detection of therapeutically relevant antibiotic resistance determinants in clinical isolates of Staphylococcus aureus. AB - An oligonucleotide microarray was constructed for the rapid and sensitive molecular detection of antibiotic resistance determinants in Staphylococcus aureus. The array is equipped with oligonucleotide capture probes for the detection of 10 clinically and therapeutically relevant antibiotic resistance genes and -mutations (mecA, aacA-aphD, tetK, tetM, vat(A), vat(B), vat(C), erm(A), erm(C), grlA-mutation) as well as several control probes. A microarray concept was established including multiplexed PCR amplification, DNA labeling, hybridization and data processing. This concept was applied to clinical Staphylococcus aureus isolates and results were concordant with those from standard genotypic and phenotypic resistance testing. Our microarray concept offers rapid and accurate identification of antibiotic resistance profiles. It is easily expandable and thus can be adapted to changing clinical and epidemiological requirements in clinical diagnosis as well as in epidemiological studies. PMID- 17123781 TI - Age related effects of pesticide exposure on neurobehavioral performance of adolescent farm workers in Brazil. AB - Adolescents (10 to 18 years old) from a rural area in the state of Rio de Janeiro, Brazil were tested using the Behavioral Assessment and Research System (BARS) to evaluate their performance. Participants were drawn from two areas--a school serving farm children (rural N=38) and a school serving children from a city within this area (urban N=28). These children were drawn from an ongoing study being carried out by Fundacao Oswaldo Cruz where rural and urban participants were matched for age, sex, and level in school. For this ongoing study, the participants had been interviewed to determine their occupational history, especially their activities that might involve exposure to pesticides. Interview responses were used to assign an exposure index to each participant. While a few BARS performance measures suggested behavioral impairments for the rural versus the urban participants, a stronger and more consistent association between BARS measures (especially impairment of tapping, digit span, and selective attention) and level of exposure to pesticides was noted when the exposure index was entered into a multiple linear regression analysis. Exposure appeared to be especially strong for the youngest participants (10-11 years old). The relationship between these behaviors and the exposure index was stronger than the relationship between behaviors and either age or sex. PMID- 17123782 TI - Dose-dependent characteristics of intravenous calcium therapy for hypocalcemic critically ill trauma patients receiving specialized nutritional support. AB - OBJECTIVE: The purpose of this investigation was to evaluate the dose-dependent characteristics of intravenous calcium gluconate therapy for hypocalcemic critically ill patients. METHODS: The dose-dependent characteristics of 2 g versus 4 g of intravenous calcium gluconate therapy were evaluated in 25 critically ill, adult multiple trauma patients with hypocalcemia. The calcium gluconate was infused at a rate of 1 g/h for both groups. Patients weighed within 90% to 120% of ideal body weight, had normal renal function, did not receive diuretic therapy, and did not have anasarca. RESULTS: Fifteen patients with mild hypocalcemia (serum ionized calcium concentration [iCa] 1-1.12 mmol/L) were given 2 g of calcium gluconate. Ten patients with moderate to severe hypocalcemia (iCa <1 mmol/L) were given 4 g. Each dosage group had a significant (P < or = 0.001) increase in iCa (from 1.07 +/- 0.05 to 1.17 +/- 0.05 mmol/L and from 0.92 +/- 0.08 to 1.16 +/- 0.11 mmol/L, respectively). Each dosage group retained about half of the dose in the exchangeable calcium space (P = NS between groups), but the higher dosage group retained significantly more elemental calcium overall (81 +/- 38 versus 201 +/- 50 mg, respectively, P < or = 001). Serum ionized calcium concentrations achieved a plateau without a further decline in iCa by 10 h after completion of the infusion for each dosage. CONCLUSION: About half of the administered elemental calcium dose was retained for each dosage group, with the higher dose (4 g) resulting in significantly more elemental calcium retention in the exchangeable calcium space. An iCa determination performed about > or =10 h after the completion of the calcium gluconate infusion should be sufficient time to ensure equilibration of iCa to assess the efficacy of the therapy. This mode of calcium therapy serves as an effective means for providing calcium to the acutely hypocalcemic, critically ill, multiple trauma patient. PMID- 17123783 TI - Cardiovascular and metabolic risk in outpatients with schizophrenia treated with antipsychotics: results of the CLAMORS Study. AB - AIM: To assess the prevalence of Coronary Heart Disease (CHD) and Metabolic Syndrome (MS) in patients treated with antipsychotics. METHODS: Retrospective, cross-sectional, multicenter study in which 117 Spanish psychiatrists (the CLAMORS Study Collaborative Group) recruited consecutive outpatients meeting DSM IV criteria for Schizophrenia, Schizophreniform or Schizoaffective Disorder, receiving antipsychotic treatment for at least 12 weeks. CHD risk was assessed by SCORE (10-year CV death) and Framingham (10-year all CHD events) function. MS was defined by at least 3 of the following components: waist circumference >102 (men)/>88 (women) cm; triglycerides > or =150 mg/dl; HDL-cholesterol <40 mg/dl (men)/<50 mg/dl (women); blood pressure > or =130/85; fasting glucose > or =110 mg/dl. RESULTS: 1452 evaluable patients (863 men, 60.9%), aged 40.7+/-12.2 years (mean+/-SD) were included. MS was present in 24.6% [23.6% (men), 27.2% (women); p=0.130)]. The overall 10-year risks were 0.9+/-1.9 (SCORE) and 7.2+/-7.6 (Framingham). 8% (95%CI: 6.5-9.5) and 22.1% (95%CI: 20.0-24.3) of patients showed a high/very high risk according to SCORE (> or =3%) and Framingham (> or =10%) function. Abdominal obesity and low HDL-cholesterol were more prevalent in women: 54.5% (95%CI: 50.2-58.9) versus 34.3% (95%CI: 31.0-37.7), and 46.1% (95%CI: 41.4) versus 28.5 (95%CI: 50.8), p<0.001 in both cases. Hypertension and hypertriglyceridemia were more prevalent in men: 59.0% (95%CI: 55.7-62.3) versus 46.0% (95%CI: 41.8-50.2), and 40.7% (95%CI: 37.2-44.2) versus 32.4 (95%CI: 28.3 36.5), p<0.01 in both cases. CONCLUSIONS: CHD risk and MS prevalences among patients with schizophrenia treated with antipsychotics were in the same range as the Spanish general population 10 to 15 years older. PMID- 17123784 TI - Adult neurogenesis and schizophrenia: a window on abnormal early brain development? AB - Adult neurogenesis is one of the most rapidly growing areas in neuroscience research and there is great interest in its potential role in the pathophysiology of psychiatric illness. In parallel with early development, adult neurogenesis occurs through the proliferation of precursor cells which migrate to specific regions and differentiate into neurons with characteristics indistinguishable from existing mature neurons. These findings have led to the re-definition of the concept of network plasticity in the adult brain, to include the formation of new neurons as well as new connections. This review examines the idea that adult neurogenesis may be disturbed in schizophrenia. We discuss evidence for abnormal mechanisms of neurogenesis and expression of developmental genes in schizophrenia, the influence of antipsychotic drugs on neurogenesis and the role of candidate schizophrenia susceptibility genes in adult neurogenesis. The recent discovery of molecular markers transiently expressed in newborn neurons within adult neurogenic brain regions could be used to probe whether neurogenesis is disturbed in schizophrenia. Adult neurogenesis could also be used as a unique tool for investigating genes involved in early brain development using post mortem brains. This is particularly relevant for brain disorders with developmental origins such as schizophrenia. PMID- 17123785 TI - COMT val108/158met genotype, cognitive function, and cognitive improvement with clozapine in schizophrenia. AB - Preliminary evidence suggests that a single nucleotide polymorphism (SNP), the val108/158met SNP, within the gene that codes for catechol-O-methyltransferase (COMT), a key enzyme involved in regulating dopamine (DA) transmission within the prefrontal cortex (PFC), is related to cognitive function in schizophrenia and cognitive improvement with atypical antipsychotic drugs (APDs). Specifically, several studies have identified an association between working memory and executive functions, and COMT val108/158met genotype in schizophrenia; although there have been several negative findings that are likely related to small sample sizes and, possibly, medication status of patients at the time of testing. The association between COMT val108/158met genotype, cognitive function, and cognitive improvement with clozapine was investigated in a relatively large prospective sample of patients with schizophrenia, most of whom were unmedicated at baseline. Patients were genotyped for the COMT val108/158met SNP after completing a cognitive battery consisting of tests of attention, working memory, verbal learning and memory, executive function, and verbal fluency at baseline and after 6 weeks and 6 months of treatment with clozapine. Consistent with several previous studies, an association between COMT genotype and tests of executive function and working memory was identified at baseline. In addition, a novel interaction between genotype and improvement on tests of attention and verbal fluency was identified. Specifically, met homozygous and val/met heterozygous patients demonstrated significantly greater improvement than val homozygous patients following 6 months of treatment with clozapine. The results are discussed in relation to previous cross-sectional studies and prospective investigations of the associations between COMT genotype, cognition, and cognitive improvement with atypical APDs in schizophrenia. PMID- 17123786 TI - Suicidality in schizophrenic patients with and without obsessive-compulsive disorder. AB - This report examines the suicidal behaviour in subjects with schizophrenia who have (N=24) and do not have comorbid Obsessive-Compulsive Disorder (OCD) (N=33). The patients with OCD-schizophrenia were more likely to have a previous history of suicidal attempts, and ideations. The number of previous suicidal attempts were significantly higher in patients with OCD-schizophrenia than in patients with non-OCD schizophrenia. The patients with a history of previous suicide attempts were more likely to have a comorbid diagnosis of OCD. Compulsive symptoms were significant predictors of suicide attempt among patients with schizophrenia. Our preliminary findings may suggest that obsessive-compulsive symptoms may account for the emergence of suicidality in patients with OCD schizophrenia. PMID- 17123787 TI - Cognitive deficits in first-episode psychosis: a population-based study in Sao Paulo, Brazil. AB - BACKGROUND: Studies conducted in high-income countries have reported significant cognitive deficits in first on set schizophrenia subjects relative to asymptotic controls, and it has been suggested that the severity of such deficits could be directly related to the duration of untreated psychosis (DUP). It is relevant to conduct similar studies in developing countries, given the supposedly better outcome for schizophrenia patients living in the latter environments. METHODS: We applied verbal fluency and digit span tests to an epidemiological-based series of patients with first-onset psychoses (n=179) recruited in the city of Sao Paulo, and compared the findings with those from non-psychotic control subjects randomly selected from the same geographical areas (n=383). RESULTS: Psychosis subjects showed lower scores on the three tests relative to controls, with greatest between-group differences for the backward digit span task (p<0.0001). There were no significant differences between subjects with affective and schizophreniform psychosis. Cognitive performance indices were negatively correlated with the severity of negative symptoms, but showed no relation to DUP. CONCLUSION: We found significant cognitive deficits in patients investigated early during the course of psychotic disorders in an environment that is distinct from those where the subjects investigated in previous studies have been drawn from. We found no support to the hypothesis of an association between greater cognitive deficits and a longer DUP. PMID- 17123789 TI - The eumelanin and pheomelanin contents in dorsal hairs of female recessive yellow mice are greater than in male. AB - BACKGROUND: The murine recessive yellow (Mc1r(e)) is a loss-of-function mutation in the receptor for alpha-melanocyte-stimulating hormone (MSH), melanocortin receptor 1 (MC1R), and produces yellow coats by inducing pheomelanin synthesis in hair follicular melanocytes. OBJECTIVE: It is not known whether the Mc1r(e) mutation affects pheomelanin synthesis in other skin sites. In this study, the eumelanin and pheomelanin contents in the epidermis and dermis as well as hairs of wild-type and mutant mice were measured. MATERIALS AND METHODS: The content of melanin was measured by high performance liquid chromatography. RESULTS: The eumelanin contents in the epidermis and dermis of newborn wild-type (Mc1r(+)/Mc1r(+)) mice (0.5, 3.5, 5.5, and 7.5 days) were much greater than those of mutant (Mc1r(e)/Mc1r(e)) mice, whereas the pheomelanin contents in the epidermis and dermis of mutant mice were much greater than those of wild-type mice. No sex differences in the contents of eumelanin and pheomalanin in the epidermis and dermis both in mutant and wild-type mice were observed. The eumelanin contents in mutant hairs (5-week-old) was much smaller than in wild type hairs, whereas the pheomelanin contents in mutant hairs was much greater than in wild-type hairs. However, the eumelanin and pheomelanin contents in mutant female hairs were greater than in male. These sex differences were not observed in wild-type mice. CONCLUSION: The Mc1r(e) gene stimulates pheomelanin synthesis in the epidermis, dermis and hair follicles. In addition, eumelanin and pheomelanin contents in Mc1r(e)/Mc1r(e) hairs may be influenced by the sex difference. PMID- 17123788 TI - Roles for IL-1 and TNFalpha in dynamic behavioral responses of Langerhans cells to topical hapten application. AB - BACKGROUND: To understand the behavioral biology of Langerhans cells (LCs), we recently recorded time-lapse images of LCs in the knock-in mice expressing the I Abeta chain tagged with the enhanced green fluorescence protein (EGFP). EGFP(+) LCs showed relatively limited motility in the steady state, whereas topical application of dinitrofluorobenzene (DNFB) markedly augmented a unique movement of dendrites characterized by rhythmic extension and retraction, termed dSEARCH, and triggered amoeba-like lateral migration of cell bodies. OBJECTIVE: To define underlying mechanisms by which hapten treatment alters LC behaviors. METHODS: The I-Abeta-EGFP mice received subcutaneous (s.c.) injection of recombinant IL-1alpha or TNFalpha (50 ng/animal) and dynamic behaviors of EGFP(+) LCs were recorded by time-lapse confocal microscopy at several time points to measure their dSEARCH activities and lateral migration. In a different set of experiments, IL-1 receptor antagonist (IL-1Ra) or soluble TNF receptor-2 (sTNFR2) (0.5 microg/animal) was s.c. injected into the ear skin 30 min before topical application of DNFB, and LC behaviors analyzed 30 h later. RESULTS: Local injection of IL-1alpha or TNFalpha induced significant, albeit modest, augmentation of both dSEARCH and lateral migration. Co-injection of TNFalpha and IL-1alpha further exacerbated motile activities in a synergistic manner by similar magnitudes observed after DNFB application. Conversely, DNFB-induced behavioral changes were inhibited completely by local injection of IL-1Ra or sTNFR2. CONCLUSION: IL-1 and TNFalpha serve as equally important mediators of hapten-induced alteration of LC behaviors. Motile activities of epidermal LCs are reprogrammed by selected cytokines known to be produced by keratinocytes under pathological conditions. PMID- 17123790 TI - Differences in blood pregnenolone and dehydroepiandrosterone levels between schizophrenia patients and healthy subjects. AB - BACKGROUND AND OBJECTIVE: Contradictory and confusing reports on serum dehydroepiandrosterone (DHEA) levels in schizophrenia led us to compare the serum concentration of its precursor, pregnenolone (PREG), between medicated schizophrenia patients and healthy subjects. The neurosteroid levels were monitored for two months and the relationship of these neurosteroids with schizophrenic symptomatology, emotional distress, and anxiety was examined. METHOD: We determined blood levels of PREG, and DHEA in 15 schizophrenia patients and 12 healthy controls at four time points: at the start of the study, after 2, 4 and 8 weeks. Analysis of covariance and canonical correlations across four time points were applied. RESULTS: Controlling for age, serum concentrations of PREG were lower, while the DHEA level and the molar ratio values of DHEA/PREG were higher in schizophrenia patients compared to healthy controls. Both levels of PREG and DHEA and their molar ratio did not change significantly during the study's period either among schizophrenia patients or healthy controls. The blood levels of PREG appear to be associated with trait-anxiety scores in the schizophrenia patients, while associations of clinical symptoms with two neurosteroids did not reach a significant level when the confounding effect of emotional distress, and anxiety scores was controlled. CONCLUSION: Low serum pregnenolone concentrations in schizophrenia appear to be associated with trait anxiety scores independent of symptoms. Further research into the role of pregnenolone in schizophrenia is warranted. PMID- 17123791 TI - The limb-girdle muscular dystrophies--diagnostic strategies. AB - The limb-girdle muscular dystrophies are a group of disorders where our understanding of their underlying molecular basis has made huge strides over the past years, revealing great heterogeneity at the clinical and molecular level. The availability of direct protein and/ or gene based approaches to diagnosis means that these disorders can now be precisely defined, and such definition of a precise diagnosis is increasingly allowing directed management for these diseases by the ability to predict specific complications such as those of the cardiac or respiratory systems. An algorithm combining clinical, biochemical and molecular testing is described which will aid precision of diagnosis and direct specific testing towards the cases most likely to benefit. This brings advantages for the patients of today in recognising the specific risks of their disorders, and in the future will be the starting point for specific gene and protein based therapies. PMID- 17123792 TI - Impaired neural development caused by inducible expression of Axin in transgenic mice. AB - Ablations of the Axin family genes demonstrated that they modulate Wnt signaling in key processes of mammalian development. The ubiquitously expressed Axin1 plays an important role in formation of the embryonic neural axis, while Axin2 is essential for craniofacial skeletogenesis. Although Axin2 is also highly expressed during early neural development, including the neural tube and neural crest, it is not essential for these processes, apparently due to functional redundancy with Axin1. To further investigate the role of Wnt signaling during early neural development, and its potential regulation by Axins, we developed a mouse model for conditional gene activation in the Axin2-expressing domains. We show that gene expression can be successfully targeted to the Axin2-expressing cells in a spatially and temporally specific fashion. High levels of Axin in this domain induce a region-specific effect on the patterning of neural tube. In the mutant embryos, only the development of midbrain is severely impaired even though the transgene is expressed throughout the neural tube. Axin apparently regulates beta-catenin in coordinating cell cycle progression, cell adhesion and survival of neuroepithelial precursors during development of ventricles. Our data support the conclusion that the development of embryonic neural axis is highly sensitive to the level of Wnt signaling. PMID- 17123793 TI - Comparisons of the foaming and interfacial properties of whey protein isolate and egg white proteins. AB - Whipped foams (10%, w/v protein, pH 7.0) were prepared from commercially available samples of whey protein isolate (WPI) and egg white protein (EWP), and subsequently compared based on yield stress (tau(0)), overrun and drainage stability. Adsorption rates and interfacial rheological measurements at a model air/water interface were quantified via pendant drop tensiometry to better understand foaming differences among the ingredients. The highest tau(0) and resistance to drainage were observed for standard EWP, followed by EWP with added 0.1% (w/w) sodium lauryl sulfate, and then WPI. Addition of 25% (w/w) sucrose increased tau(0) and drainage resistance of the EWP-based ingredients, whereas it decreased tau(0) of WPI foams and minimally affected their drainage rates. These differing sugar effects were reflected in the interfacial rheological measurements, as sucrose addition increased the dilatational elasticity for both EWP-based ingredients, while decreasing this parameter for WPI. Previously observed relationships between tau(0) and interfacial rheology did not hold across the protein types; however, these measurements did effectively differentiate foaming behaviors within EWP-based ingredients and within WPI. Interfacial data was also collected for purified beta-lactoglobulin (beta-lg) and ovalbumin, the primary proteins of WPI and EWP, respectively. The addition of 25% (w/w) sucrose increased the dilatational elasticity for adsorbed layers of beta lg, while minimally affecting the interfacial rheology of adsorbed ovalbumin, in contrast to the response of WPI and EWP ingredients. These experiments underscore the importance of utilizing the same materials for interfacial measurements as used for foaming experiments, if one is to properly infer interfacial information/mechanisms and relate this information to bulk foaming measurements. The effects of protein concentration and measurement time on interfacial rheology were also considered as they relate to bulk foam properties. This data should be of practical assistance to those designing aerated food products, as it has not been previously reported that sucrose addition improves the foaming characteristics of EWP-based ingredients while negatively affecting the foaming behavior of WPI, as these types of protein isolates are common to the food industry. PMID- 17123794 TI - [Neonatal splenic injury: a difficult diagnosis]. AB - Neonatal splenic injury is a rare but serious condition, due to the risk of haemorrhagic shock. We report on the case of a newborn infant with a neonatal respiratory distress that first evoked materno-fetal infection. Clinical deterioration, with anemia and abdominal distension, led then to the proper diagnosis. Dystocia seems to be the most likely cause of the splenic rupture in this report. Medical treatment is advocated as first line, while surgical treatment may be necessary in some cases. In the case surgery is inevitable, a conservative approach is preferable. PMID- 17123795 TI - [Neonatal subcutaneous fat necrosis with hypercalcemia: efficiency of a low dose of corticosteroids]. AB - We report the case of a newborn with macrosomia, extensive subcutaneous fat necrosis and symptomatic hypercalcemia. Low doses of prednisone were efficient, while dietary intervention, hyperhydratation and furosemide were not. Treatment of hypercalcemia in this specific neonatal condition are discussed. PMID- 17123796 TI - The effect of suction during die fill on a rotary tablet press. AB - Die fill on a rotary tablet press involves complex powder flow phenomena. Conventional techniques for measuring flowability do not normally provide information that is directly relevant to the design of powder feed systems or to the selection of press parameters for the die filling process. Sinka et al. [I.C. Sinka, L.C.R. Schneider, A.C.F. Cocks, Measurement of the flow properties of powders with special reference to die fill, in: International Journal of Pharmaceutics 280 (1-2) (2004) 27-38] used an experimental shoe-die system to characterise the flow behaviour of pharmaceutical powders. A rigorous data analysis procedure was developed by Schneider et al. [L.C.R. Schneider, I.C. Sinka, A.C.F. Cocks, Characterisation of the flow behaviour of pharmaceutical powders using a model die-shoe filling system, in: Powder Technology (in press)] to evaluate the experimental results, however, when scaling the results to a rotary tablet press, the die fill efficiency was underpredicted by a factor of approximately 2, because the experimental system did not capture major features of the rotary press flow process. The suction effect, whereby the lower punch is moved downwards while the top of the die is exposed to powder in the feed system, is a key element of the process. In this note we describe the development of a model shoe-die system that allows the effect of suction to be investigated. The results demonstrate the improvement offered by suction and illustrate how a fundamental understanding of die fill phenomena could assist the selection of process parameters to maximise the operational speed of a rotary press. PMID- 17123797 TI - Nuclear targeting of non-viral gene carriers using psoralen-nuclear localization signal (NLS) conjugates. AB - A nuclear localization signal was non-covalently attached to DNA for the purpose of enhancing transfection efficiencies of non-viral gene carriers. Psoralen, a nucleic acid-intercalating agent, was chemically attached to a signal peptide. The conjugate spontaneously intercalated into DNA and then poly(ethyleneimine) [PEI] was added to prepare a DNA/PEI complex containing the signal peptide moieties. The existence of the conjugate did not alter the complexation process between DNA and PEI, which was confirmed by dynamic light scattering. The conjugate was slowly released from the DNA/PEI complex for 24h, while a burst release was examined when the conjugated was added to DNA without PEI. The complex containing a signal peptide moiety increased transfection efficiencies on COS-1 cells, compared to a mutant signal peptide or a control. Cytotoxicity of the conjugate slowly increased as the amount of the conjugate increased, however, the cytotoxic effect of the conjugate was not significant at the effective concentration of the conjugate for transfections. Therefore, the psoralen-nuclear localization signal is expected to be a potent transfection enhancing agent without a covalent modification of transgenes. PMID- 17123798 TI - Use of calcined Mg-Al-hydrotalcite to enhance the stability of celecoxib in the amorphous form. AB - The use of compound in amorphous forms is a promising approach to improve solubility of poorly water-soluble drugs. However, during storage, amorphous state can spontaneously transform to the lower energy crystalline form and this is a limiting factor for a large commercial use of drugs. In this paper, calcined hydrotalcite was employed to support amorphous celecoxib and several preparations at different weight drug/carrier ratio were prepared. Solubility of celecoxib from the prepared systems was evaluated and its physical stability during storage at different conditions was examined as well. The results show that HTlc-calc can be used as a support of amorphous celecoxib with consequent improvement of drug solubility and physical stability. PMID- 17123799 TI - Tetronic micellization, gelation and drug solubilization: Influence of pH and ionic strength. AB - The aim of this work was to gain an insight into the self-associative processes and drug solubilization ability of a Tetronic variety, T904 (4 x 15 EO units; 4 x 17 PO units; HLB 15), in aqueous media covering the physiological range of pH and ionic strength, applying isoperibol microcalorimetry, transmission electronic microscopy (TEM), dynamic light scattering (DLS), oscillatory rheometry, and drug diffusion experiments. T904 shows two pK(a) (pK(a1)=4.0 and pK(a2)=7.9) and, at pH<5.8, the diprotonated form predominates over the non-protonated one. Deprotonization of the central diamine group is a required condition for micellization, which is an endothermic entropy-driven process owing to hydrophobic interactions between the PPO chains. As the pH of the solutions decreases, the coulombic repulsions among the positively charged amine groups make the aggregation more difficult, raising the critical micellar concentration (CMC) and decreasing the size of the micelles. The changes in the conformation and hydrophilicity of the Tetronic were reflected in its gelation temperature (around 30 degrees C at neutral-alkaline pH; no gelation at pH<2) and solubilization capacity for griseofulvin (2-fold greater at neutral-alkaline pH than at pH<2) and rate of diffusion (slower at pH 7.4). Such alterations in self assembly are relevant when using Tetronic in the design of drug delivery systems. PMID- 17123800 TI - In vitro and in vivo characterization of a combination chemotherapy formulation consisting of vinorelbine and phosphatidylserine. AB - The purpose of these studies was to design an intravenous drug formulation consisting of two active agents having synergistic in vitro activity. Specifically, we describe a novel drug combination consisting of a cytotoxic agent (vinorelbine) with an apoptosis-inducing lipid (phosphatidylserine, PS). In vitro cytotoxicity screening of PS and vinorelbine, alone and in combination, against human MDA435/LCC6 breast cancer and H460 lung cancer cells was used to identify the molar ratio of these two agents required for synergistic activity. PS and vinorelbine were co-formulated in a lipid-based system at the synergistic molar ratio and the pharmacokinetic and antitumor characteristics of the combination assessed in mice bearing H460 tumors. The cytotoxicity of the lipid, and the synergy between the lipid and vinorelbine, were specific to PS; these effects were not observed using control lipids. A novel formulation of PS, incorporated as a membrane component in liposomes, and encapsulating vinorelbine using a pH gradient based loading method was developed. The PS to vinorelbine ratio in this formulation was 1/1, a ratio that produced synergistic in vitro cytotoxicity over a broad concentration range. The vinorelbine and PS dual-agent treatment significantly delayed the growth of subcutaneous human H460 xenograft tumors in Rag2M mice compared to the same dose of free vinorelbine given alone or given as a cocktail of the free vinorelbine simultaneously with empty PS containing liposomes. These studies demonstrate the potential to develop clinically relevant drug combinations identified using in vitro drug-drug interactions combined with lipid-based delivery systems to co-formulate drugs at their synergistic ratios. PMID- 17123801 TI - An integrated early formulation strategy--from hit evaluation to preclinical candidate profiling. AB - The selection of a suitable vehicle for preclinical compound profiling is a very important task during the early developmental phases to ensure the quality of candidates and the speed of compound progression. Apart from biopharmaceutical and pharmaceutical technical considerations, i.e. solubility/dissolution improvement or route of application, other aspects have to be taken into account, as well: (i) availability and quality of the compound, (ii) tolerability of the vehicle in the selected animal model, (iii) developmental possibilities, i.e. whether the formulation can be transformed into a clinical one. The approach described in this paper is based on results of team collaboration between functions involved in the conduct of animal experiments (Pharmacology, Pharmacokinetics, Toxicology, and Pharmaceutical Sciences). Very early in vivo studies should be performed with dissolved API as available information on solid state characteristics is usually limited at this time. Later studies should be performed with developable formulations, taking into consideration pharmacological, toxicological, and pharmaceutical requirements. At this stage, delivery strategies (i.e. advanced formulations and/or alternative routes of administration) should be considered, as well. In addition, a minimum level analytical characterization of compounds and formulations used in animal studies is required to explain unexpected results. PMID- 17123802 TI - pH-responsive polymeric micelles of poly(ethylene glycol)-b poly(alkyl(meth)acrylate-co-methacrylic acid): influence of the copolymer composition on self-assembling properties and release of candesartan cilexetil. AB - The objective of the present study was to investigate the influence of chemical structure and molecular weight of pH-sensitive block copolymers on their self assembling properties, the loading and the release of candesartan cilexetil (CDN). Block copolymers of poly(ethylene glycol) and t-butyl methacrylate, iso butyl acrylate, n-butyl acrylate or propyl methacrylate were synthesized by atom transfer radical polymerization. pH-sensitivity was obtained by hydrolysis of t butyl groups. The poorly water-soluble drug CDN was incorporated in the micelles and the in vitro drug release was evaluated as a function of pH. The critical aggregation concentration of hydrolyzed copolymers (pK(a)=6.2-6.6) was higher compared to the unhydrolyzed ones. Dynamic light scattering studies and atomic force microscopy images revealed uniform size micelles with aggregation numbers ranging from 60 to 160. The entrapment efficiency of CDN was generally found to be above 90%, with drug loading levels reaching approximately 20% (w/w). Differential scanning calorimetry studies showed the amorphous nature of entrapped CDN. The release of CDN from pH-sensitive micelles was triggered upon an increase in pH from 1.2 to 7.2. These findings suggest that the PEG-b poly(alkyl(meth)acrylate-co-methacrylic acid)s can self-assemble to form micelles which exhibit high loading capacities for CDN and release the drug in a pH dependent fashion. PMID- 17123803 TI - Interindividual differences in anticancer drug cytotoxicity in primary human glioblastoma cells. AB - Glioblastoma multiforme is a malignant astrocytic tumor characterized by rapid growth, extensive invasiveness and high vascularity. Despite advances in surgical techniques and in the development of new protocols in radio- and chemotherapy, the prognosis for patients suffering from this malignancy remains poor. Since the clinical response to chemotherapy varies greatly owing to different interindividual gene expression profiles, it would be of considerable interest to develop an in vitro model able to evaluate anticancer drug toxicity and the effectiveness of therapeutic strategies on cells obtained from individual patients. In the protocol for obtaining primary cultures of glioblastoma cells described in this report, a confluent monolayer of cells can be obtained within 1 or 2 weeks. A complementary immunocytochemical assay using glial fibrillary acidic protein (GFAP) to reliably mark glial cells confirms the glial origin of the cultured cells. A cytotoxicity test based on mitochondrial activity is then used to evaluate in vitro drug efficacy. Cell dedifferentiation as evidenced by loss of GFAP expression after a few passages requires determination of drug toxicity before the fourth passage. Data show a wide range of response to temozolomide (1000 microM) after 72 h with 24-81% cell death depending on patients. Results presented confirm the heterogeneity of response to anticancer drugs between the patients and methods described allow to carry out cytotoxicity studies in order to determine the individualized most effective treatment. PMID- 17123804 TI - Bioassay by intratracheal instillation for detection of lung toxicity due to fine particles in F344 male rats. AB - We have established and documented an in vivo bioassay for detection of hazards with intratracheally instilled fine particles, which can be used for risk assessment of toxicity of materials inhaled into deep lung tissue of humans (Yokohira et al. Establishment of a bioassay system for detection of lung toxicity due to fine particle instillation: sequential histopathological changes with acute and subacute lung damage due to intratracheal instillation of quartz in F344 male rats. J Toxicol Pathol 2005;18:13-8). For validation we here examined toxicity of fine particles from quartz, hydrotalcite, potassium octatitanate, palladium oxide and carbon black with this bioassay. A total of 108, 10-week-old F344/DuCrj male rats were randomly divided into 8 groups. Groups 1 to 5 underwent intratracheal instillation of the 5 test particles (4 mg/rat) suspended in 0.2 ml vehicle (saline or 10% propylene glycol and 1% sodium carboxymethyl cellulose in saline: PG-CMC) with a specially designed aerolizer, and subgroups of 7 rats were killed on Days 1 and 28 thereafter. Groups 6 and 7 similarly were exposed to saline and PG-CMC, respectively, as vehicle controls, while group 8 was maintained untreated. Using histopathological changes and immunohistochemically assessed bromodeoxyuridine (BrdU) labeling indices, inducible nitric oxide synthase (iNOS) and matrix metalloproteinase-3 (MMP-3) levels as end points, the quartz treated group exhibited high toxicity, while the values for the other particle-treated groups pointed to only slight effects. Although additional efforts are needed to establish advantages and disadvantages with our bioassay, models featuring intratracheal instillation clearly can be useful for detection of acute or subacute lung toxicity due to inhaled fine particles by using histopathological scoring and markers like BrdU and iNOS for screening purposes in short-term studies. PMID- 17123805 TI - Ablation of vitamin D signaling rescues bone, mineral, and glucose homeostasis in Fgf-23 deficient mice. AB - To explore further the role of the vitamin D axis for fibroblast growth factor-23 (FGF23) signaling, we mated Fgf-23 deficient (Fgf-23(-/-)) mice and vitamin D receptor (VDR) mutant mice with a non-functioning VDR. To prevent secondary hyperparathyroidism in VDR and compound mutant mice, all mice were kept on a rescue diet enriched with calcium, phosphorus, and lactose. Consistent with previous findings, Fgf-23(-/-) animals showed hypercalcemia, hyperphosphatemia, growth retardation, ectopic calcifications, severe osteoidosis, skin atrophy, and renal dysfunction. In addition, here we describe that Fgf-23(-/-) mice are hypoglycemic, and have profoundly increased peripheral insulin sensitivity and improved subcutaneous glucose tolerance, but normal renal expression of the aging suppressor gene Klotho. Although VDR and double mutants on the rescue diet still had moderately elevated parathyroid hormone serum levels and lower bone mineral density compared to wild-type mice, double mutant mice were normocalcemic and normophosphatemic, and had normal body weight, normal renal function, and no ectopic calcifications. Ablation of vitamin D signaling in compound mutants also normalized subcutaneous glucose tolerance tests and insulin secretory response. In conclusion, our results indicate that the alterations in mineral and carbohydrate metabolism present in Fgf-23(-/-) mice require an intact vitamin D signaling pathway. PMID- 17123806 TI - Osteopontin promotes pathologic mineralization in articular cartilage. AB - Calcium pyrophosphate dihydrate (CPPD) crystals are commonly found in osteoarthritic joint tissues, where they predict severe disease. Unlike other types of calcium phosphate crystals, CPPD crystals form almost exclusively in the pericellular matrix of damaged articular cartilage, suggesting a key role for the extracellular matrix milieu in their development. Osteopontin is a matricellular protein found in increased quantities in the pericellular matrix of osteoarthritic cartilage. Osteopontin modulates the formation of calcium containing crystals in many settings. We show here that osteopontin stimulates ATP-induced CPPD crystal formation by chondrocytes in vitro. This effect is augmented by osteopontin's incorporation into extracellular matrix by transglutaminase enzymes, is only modestly affected by its phosphorylation state, and is inhibited by integrin blockers. Surprisingly, osteopontin stimulates transglutaminase activity in cultured chondrocytes in a dose-responsive manner. As elevated levels of transglutaminase activity promote extracellular matrix changes that permit CPPD crystal formation, this is one possible mechanism of action. We demonstrate the presence of osteopontin in the pericellular matrix of chondrocytes adjacent to CPPD deposits and near active transglutaminases. Thus, osteopontin may play an important role in facilitating CPPD crystal formation in articular cartilage. PMID- 17123807 TI - The consequence of PRELP overexpression on skin. AB - PRELP is a member of the small leucine-rich repeat proteoglycan family that is abundantly expressed in many cartilages compared to other connective tissues. To study the consequence of PRELP overexpression in tissues where it is normally expressed at low abundance, transgenic mice were generated in which the human PRELP transgene was placed under control of the CMV promoter. A connective tissue phenotype was observed in the skin, where the organization of collagen fibrils in the dermis was perturbed and the thickness of the hypodermal fat layer was diminished. PMID- 17123808 TI - Fatty acids and sleep in depressed inpatients. AB - Sleep disturbances belong to the most frequent symptoms of depression. Low concentrations of n-3-fatty acids might represent one determinant within that process. Therefore, the aim of the study was to examine the relationships between serum fatty acid concentrations and severity of sleep disturbances in depressives. Serum fatty acids were measured gaschromatographically in 118 depressive inpatients (51 males; 67 females; age 45.4+/-12.0 years), divided into subgroups according to three degrees of sleep disturbances (BDI-item). At admission and discharge, we found significant negative correlations between the degree of sleep disturbances and fatty acid concentrations (myristic, palmitic, palmitoleic, oleic, linoleic, eicosadienoic and docosahexaenoic acid). At both assessments palmitoleic and eicosadienoic acids had the strongest connections with sleep performance. Palmitoleic and oleic acid seem to be especially important for sleep disorders, may be due to their function as precursors of the sleep inducing oleamide. Linoleic and eicosadienoic acid could be helpful for maintaining sleep because they are precursors of the sleep mediator PGD2. In contrast to our hypothesis, there is not only a significant lack of n-3 fatty acids but also of special monoenoic and n-6 fatty acids in sleep-disturbed depressives. PMID- 17123809 TI - Multilocus sequence analysis of Fusarium pseudograminearum reveals a single phylogenetic species. AB - Fusarium pseudograminearum causes crown rot of wheat in Australia and most other wheat growing regions, but its evolutionary history is largely unknown. We demonstrate for the first time that F. pseudograminearum is a single phylogenetic species without consistent lineage development across genes. Isolates of F. pseudograminearum, F. graminearum sensu lato, and F. cerealis, were collected from four countries and four single copy, nuclear genes were partially sequenced, aligned with previously published sequences of these and related species, and analysed by maximum parsimony and Bayesian inference. Evolutionary divergence varied between genes, with high phylogenetic incongruence occurring between the gene genealogies. The absence of geographic differentiation between isolates indicates that the introduction of new fungal strains to a region has the potential to introduce new pathogenic and toxigenic genes into the native population through sexual recombination. PMID- 17123810 TI - Fruiting body-guided molecular identification of root-tip mantle mycelia provides strong indications of ectomycorrhizal associations in two species of Sistotrema (Basidiomycota). AB - Fruiting body guided sequence analysis of mycorrhizal root-tip mycelia is a powerful yet relatively sparsely explored method for species-level identification of mycorrhizal fungi. It is used in this study to indicate mycorrhizal associations in the corticioid (resupinate) genus Sistotrema of the cantharelloid clade through phylogenetic analysis of the ITS and nuLSU rDNA regions of two spatiotemporally co-occurring Sistotrema fruiting bodies and ectomycorrhizal root tips. The genus Sistotrema is confirmed to be polyphyletic, and the mycorrhizal species form a strongly supported monophyletic clade together with the stipitate genus Hydnum. The remaining lineages of Sistotrema may well be saprotrophic, the nutritional mode traditionally attributed to the genus, but the phylogenetic analyses show that they should be excluded from Sistotrema. The cantharelloid clade contains several mycorrhizal genera, but no symbiotic associations have previously been demonstrated for Sistotrema. PMID- 17123811 TI - Phosphonoacetic acid utilization by fungal isolates: occurrence and properties of a phosphonoacetate hydrolase in some penicillia. AB - Among a collection of 18 fungal strains representing eight genera, only two strains (Penicillium oxalicum and P. minioluteum) were capable of growth on phosphonoacetic acid as sole phosphorous source. Enrichment liquid cultures in minimal medium with the compound as the only P-source selected four isolates, that were also identified as Penicillium spp. Phosphonoacetate metabolism did not lead to extracellular release of inorganic phosphate. In all cases phosphonoacetate hydrolase activity was detected in partially purified extracts, and a protein of the expected molecular mass reacted with polyclonal antibodies raised against the enzyme from P. oxalicum. There was no relation between phosphonoacetate hydrolase specific activity and growth rate or yield. Phosphonoacetic acid was the inducer of the hydrolase, independently of the concurrent availability of inorganic phosphate. Notwithstanding this, the utilization of the phosphonate was significantly inhibited in the presence of phosphate, suggesting an interference of the latter with phosphonoacetic acid uptake. PMID- 17123812 TI - Forest stand age and the occurrence of chanterelle (Cantharellus) species in Oregon's central Cascade Mountains. AB - We describe watershed-scale habitat associations of three Cantharellus species with respect to stand age. During the 1998 autumn fruiting season we collected chanterelle sporocarps from 18 forest stands in and adjacent to the H.J. Andrews experimental forest in the central Cascade Mountains of Oregon. Sampled stands represented two age categories: old growth ( approximately 350+ y) and 40-60-y old second growth naturally regenerated from clear-cut harvest. Old growth and second growth stands were spatially paired to reduce the chance of spurious habitat relationships caused by unmeasured correlated variables. We found stand age to be a good predictor of the distribution of C. subalbidus and C. formosus, but only marginally useful for predicting the occurrence of C. cascadensis. The odds that a randomly located chanterelle sporocarp will be C. subalbidus, compared to other chanterelles, are 3-23.5 times higher in old growth than in second growth. Alternatively, there is only a 4-38% chance that a randomly located sporocarp will be C. formosus in old growth. C. cascadensis was found to be uncommon throughout the study area and showed no significant habitat associations. The abundance of C. cascadensis increased substantially with decreasing elevation indicating that landscape features other than stand age may be more useful in predicting its occurrence. PMID- 17123813 TI - Cantharellus pleurotoides, a new and unusual basidiomycete from Guyana. AB - Cantharellus pleurotoides sp. nov. (Cantharellaceae, Cantharellales, Basidiomycota) is described from the Pakaraima Mountains of Guyana, occurring in rainforests dominated by ectomycorrhizal Dicymbe spp. (Caesalpiniaceae). This fungus is singular among Cantharellus species described worldwide in possessing a pleurotoid basidioma. Macromorphological, micromorphological, and habitat data are provided for the new species. PMID- 17123814 TI - Particleboard manufacturing: an innovative way to recycle paper sludge. AB - This paper presents the results on a study to use paper mill sludge for particleboard production. Single-layer board and three-layer board, with paper sludge on the surface, were fabricated. Four levels of mixing ratios of paper sludge to wood particles (0:100, 15:85, 30:70, and 45:55) were used. The boards were produced with 3% and 4% methylene diphenyl diisocyanate (MDI), and 10% and 12% urea-formaldehyde (UF) adhesives. The bending and shear strengths, water absorption, and thickness swelling of the boards were investigated. The results indicated that the mechanical properties of the boards were negatively affected by the paper sludge amount. Overall, UF-bonded particleboards gave superior mechanical performance, water resistance, and thickness swell than MDI-bonded particleboards. The strengths of the UF-bonded board decreased much more than those of MDI-bonded board as paper sludge content increased. The three-layer boards made from 15% paper sludge with 12% UF satisfied fully the minimum requirements set by EN, ASTM D 1037-99, and ANSI A208.1 standards for general uses. PMID- 17123815 TI - Paenibacillus campinasensis BL11: a wood material-utilizing bacterial strain isolated from black liquor. AB - In order to search for new thermophilic microorganisms and their enzymes, bacterial strains from black liquor of brownstock at washing stage of kraft pulping process were screened. Therein a multiple glycosyl hydrolase-producing strain, BL11, was isolated as a dominant species in the xylan-degrading bacterial population and identified as Paenibacillus campinasensis. The bacterial strain used all kinds of saccharides and polysaccharides, except lignin as carbon source and produced multiple extracellular polysaccharide-degrading enzymes, including one xylanase (41 kDa), three cellulases (42, 57 and 86 kDa), one pectinase (28 kDa) and one cyclodextrin glucanotransferase (38 kDa). P. campinasensis BL11 lacked lipase and protease activities and was able to grow over a wide range of pH, but it particularly grew well around neutral pH at 55 degrees C. Based on its physiological characteristics, it has strong potential for industrial application and bioresource utilization. PMID- 17123816 TI - Phytosterols: applications and recovery methods. AB - Phytosterols, or plant sterols, are compounds that occur naturally and bear close structural resemblance to cholesterol, but have different side-chain configurations. Phytosterols are relevant in pharmaceuticals (production of therapeutic steroids), nutrition (anti-cholesterol additives in functional foods, anti-cancer properties), and cosmetics (creams, lipstick). Phytosterols can be obtained from vegetable oils or from industrial wastes, which gives an added value to the latter. Considerable efforts have been recently dedicated to the development of efficient processes for phytosterol isolation from natural sources. The present work aims to summarize information on the applications of phytosterols and to review recent approaches, mainly from the industry, for the large-scale recovery of phytosterols. PMID- 17123817 TI - Synthesis of pyochelin-norfloxacin conjugates. AB - Using synthetic functionalized analogues of pyochelin, a siderophore common to several pathogenic Pseudomonas and Burkholderia species, four fluoroquinolone pyochelin conjugates were efficiently synthesized and evaluated for their biological activities. PMID- 17123818 TI - Synthesis and antimalarial activity of new 1,12-bis(N,N'-acetamidinyl)dodecane derivatives. AB - Amidoxime and O-substituted derivatives of the bis-alkylamidine 1,12-bis(N,N' acetamidinyl)dodecane were synthesized and evaluated as in vitro and in vivo antimalarial prodrugs. The bis-O-methylsulfonylamidoxime 8 and the bis oxadiazolone 9 derivatives show relatively potent antimalarial activity after oral administration. PMID- 17123819 TI - A scale to monitor progression and treatment of mitochondrial disease in children. AB - Mitochondrial diseases affect all age groups, but those with childhood onset often seem to experience the greatest burden of disability. In some paediatric patients this can be explained by a cumulative disability acquired over many years. In others, additional factors, including the nature and severity of the molecular defect, must be considered. To date, no large-scale studies have attempted to document the natural history of paediatric mitochondrial disease. This is in part at least, because no assessment tool has been available to plot the temporal course of a disease with such a diverse clinical spectrum. This paper describes how a practical and semi-quantitative rating scale has been devised for children with mitochondrial disease, the Newcastle paediatric mitochondrial disease scale (NPMDS). The scale is multi-dimensional and reproducible, offering a tool through which mitochondrial disease progression can be objectively monitored. We anticipate that use of this tool will facilitate both longitudinal natural history studies and the assessment of future therapeutic interventions. PMID- 17123820 TI - Synthesis of 11C-labelled (R)-OHDMI and CFMME and their evaluation as candidate radioligands for imaging central norepinephrine transporters with PET. AB - (R)-1-(10,11-Dihydro-dibenzo[b,f]azepin-5-yl)-3-methylamino-propan-2-ol ((R) OHDMI) and (S,S)-1-cyclopentyl-2-(5-fluoro-2-methoxy-phenyl)-1-morpholin-2-yl ethanol (CFMME) were synthesized and found to be potent inhibitors of norepinephrine reuptake. Each was labelled efficiently in its methyl group with carbon-11 (t(1/2)=20.4 min) as a prospective radioligand for imaging brain norepinephrine transporters (NET) with positron emission tomography (PET). The uptake and distribution of radioactivity in brain following intravenous injection of each radioligand into cynomolgus monkey was examined in vivo with PET. After injection of (R)-[(11)C]OHDMI, the maximal whole brain uptake of radioactivity was very low (1.1% of injected dose; I.D.). For occipital cortex, thalamus, lower brainstem, mesencephalon and cerebellum, radioactivity ratios to striatum at 93 min after radioligand injection were 1.35, 1.35, 1.2, 1.2 and 1.0, respectively. After injection of [(11)C]CFMME, radioactivity readily entered brain (3.5% I.D.). Ratios of radioactivity to cerebellum at 93 min for thalamus, occipital cortex, region of locus coeruleus, mesencephalon and striatum were 1.35, 1.3, 1.3, 1.2 and 1.2, respectively. Radioactive metabolites in plasma were measured by radio HPLC. (R)-[(11)C]OHDMI represented 75% of plasma radioactivity at 4 min after injection and 6% at 30 min. After injection of [(11)C]CFMME, 84% of the radioactivity in plasma represented parent at 4 min and 20% at 30 min. Since the two new hydroxylated radioligands provide only modest regional differentiation in brain uptake and form potentially troublesome lipophilic radioactive metabolites, they are concluded to be inferior to existing radioligands, such as (S,S) [(11)C]MeNER, (S,S)-[(18)F]FMeNER-D(2) and (S,S)-[(18)F]FRB-D(4), for the study of brain NETs with PET in vivo. PMID- 17123822 TI - Calibration of a 137Cs gamma-ray beam irradiator using large size chambers. AB - A (137)Cs gamma-ray beam irradiator has been calibrated in terms of air kerma using large size chambers. The available air-kerma rates range between 1.8 microGy/h (0.2 mR/h) and 5.3 mGy/h (0.6R/h). Large-volume chambers were used to characterize the source in terms of the radiation quantity air kerma (and exposure). Two types of chambers with significantly different characteristics and energy responses were used. This work shows that very good agreement can be obtained between the measurements performed with such different types of chambers. An agreement of 0.3% is observed between chambers even for the lowest air-kerma rates measured. PMID- 17123821 TI - Design, synthesis, and antiproliferative and CDK2-cyclin a inhibitory activity of novel flavopiridol analogues. AB - The design and synthesis of a small library of 8-amidoflavone, 8 sulfonamidoflavone, 8-amido-7-hydroxyflavone, and heterocyclic analogues of flavopiridol is reported. The potential activity of these compounds as kinase inhibitors was evaluated by cytotoxicity studies in MCF-7 and ID-8 cancer cell lines and inhibition of CDK2-Cyclin A enzyme activity in vitro. The antiproliferative and CDK2-Cyclin A inhibitory activity of these analogues was significantly lower than the activity of flavopiridol. Molecular docking simulations were carried out and these studies suggested a different binding orientation inside the CDK2 binding pocket for these analogues compared to flavopiridol. PMID- 17123823 TI - Natural radioactivity measurement in pumice samples used raw materials in Turkey. AB - The activity concentrations of 232Th, 226Ra, and 40K in different pumice samples have been determined by high-resolution gamma-ray spectrometry using a 110% HpGe detector. The radium equivalent activities (Raeq), external hazard index (Hex), and internal hazard index (Hin) associated with the natural radionuclides and representative level index (Igamma r) are calculated to assess the radiation hazard of the natural radioactivity in the pumice samples. The mean values of the measured radioactivity concentrations of 232Th, 226Ra, and 40K for pumice samples from the region of lakes (ROL) are 232.4+/-8.0, 196.9+/-7.8, and 1325.8+/-20.4 Bqkg(-1) and for pumice samples from Cukurova region (CR) 16.3+/-4.0, 16.1+/-4.9, and 479.7+/-170.4 Bqkg(-1), respectively. The calculated Raeq values vary from 435.9+/-12.5 to 883.6+/-41.5 Bqkg(-1) with a mean of 630.9+/-20.2 Bqkg(-1) for the ROL samples and from 49.7+/-3.3 to 101.9+/-7.2 Bqkg(-1) with a mean of 76.3+/ 23.7 Bqkg(-1) for the CR samples. For the ROL samples, Raeq are above the limit of 370 Bqkg(-1), equivalent to external gamma dose of 1.5 mSv yr(-1), recommended for the safe use of construction materials by NEA-OECD, while for the CR samples, Raeq values are lower than the limit. PMID- 17123824 TI - Root phospholipids in Azospirillum-inoculated wheat seedlings exposed to water stress. AB - Azospirillum-plant association is accompanied by biochemical changes in roots which, in turn, promote plant-growth and tolerance to water stress. To shed light on the possible factors underlying these effects, roots from Azospirillum brasilense Sp245-inoculated Triticum aestivum seedlings growing in darkness under osmotic stress were analyzed for phospholipid (PL) composition, fatty acid (FA) distribution profiles and degree of unsaturation of the major PL classes. Azospirillum inoculation diminished ion leakage and increased 2,3,5 tripheniltetrazolium reducing ability in roots of well irrigated and water stressed wheat seedlings. Total root PL content remained unaltered in all treatments. Six PL classes were detected, phosphatidylcholine (PC) and phosphatidylethanolamine (PE) comprising over 80% of the total. While water stress increased PC content and diminished that of PE, none of these changes were observed either under Azospirillum inoculation alone or when both treatments were combined. The major FAs found in both PC and PE were 16:0, 18:0, 18:1, 18:2, and 18:3. Higher PC and lower PE unsaturation than in well irrigated controls were observed in roots from Azospirillum-inoculated, water-stressed seedlings. Azospirillum inoculation could contribute to protect wheat seedlings from water stress through changes in the FA distribution profiles of PC and PE major root phospholipids. PMID- 17123825 TI - Leaf-targeted phytochelatin synthase in Arabidopsis thaliana. AB - One of the key steps in developing transgenic plants for the phytoremediation of metal containing soils is to develop plants that accumulate metals in the aerial tissues. With the goal of changing the distribution of phytochelatin (PC) dependent cadmium accumulation from roots to the leaves, the phytochelatin synthase (PCS) deficient cad1-3 mutant and wild type (Col-0) Arabidopsis plants were transformed with an Arabidopsis phytochelatin synthase (AtPCS1) under the control of a leaf-specific promoter. Three independent transformant lines from each genetic background were chosen for further analysis and designated cad-PCS and WT-PCS. PCS activity in the cadPCS lines was restored in the leaves, but not in the roots. Additionally, when whole plants were treated with cadmium, PCs were found only in the leaves of cad-PCS plants. Although the inserted AtPCS1 gene was leaf-specific, cad-PCS lines showed an overall decrease in cadmium toxicity evidenced by a partial amelioration of the "brown-root" phenotype and root growth was restored to wild type levels when treated with cadmium and arsenate. WT-PCS lines showed an increase in leaf PCS activity but had only wild type PC levels. In addition, cadmium uptake studies indicated that there was no difference in cadmium accumulation among all types tested. So, while we were able to protect the plants against cadmium by expressing PC synthase only in the leaves, we were not able to limit cadmium accumulation to aerial tissues. PMID- 17123826 TI - Photon emissions from rice cells elicited by N-acetylchitooligosaccharide are generated through phospholipid signaling in close association with the production of reactive oxygen species. AB - Biophotons are ultraweak light emissions from biochemical reactions in a living body. They increase in suspension-cultured rice (Oryza sativa L.) cells when elicited by N-acetylchitooligosaccharide. Biochemical analyses were undertaken to investigate the relationship between disease response and biophotons in order to clarify the emission mechanism of biophotons caused by this elicitor. Photon emissions induced by N-acetylchitohexaose were suppressed when cells were pretreated with the reactive oxygen species (ROS)-generating inhibitors: pyrocatechol-3,5-disulfonic acid disodium salt (Tiron); diphenylene iodonium (DPI); and salicylhydroxamic acid (SHAM). Conversely, exogenously applied ROS (superoxide and hydrogen peroxide) were able to induce photon emissions. The effects of protein phosphorylation (K-252a) and the Ca(2+) signaling inhibitors, ethylene glycol-bis(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) and LaCl(3), caused photon emissions to decrease. It is clear that photon emissions from rice cells elicited by N-acetylchitohexaose are closely associated with the ROS-generating system, and are regulated by Ca(2+) signaling and protein phosphorylation. Exogenously applied phosphatidic acid (PA), the second messenger in the signal transduction of disease response, raised photon emissions in rice cells. Comparisons of photon emissions from PA and N-acetylchitohexaose regarding time courses, spectral compositions, and the inhibition ratios of several inhibitors, as well as a loss- and gain-of-function assay using the protein synthesis inhibitor cycloheximide (CHX) and PA, showed the possibility that photon emissions from rice cells elicited by N-acetylchitooligosaccharide were generated through PA, an intermediate of phospholipid signaling. PMID- 17123828 TI - Detection and identification of Bacteriovorax stolpii UKi2 Sphingophosphonolipid molecular species. AB - Bacteriovorax stolpii is a predator of larger gram-negative bacteria and lives as a parasite in the intraperiplasmic space of the host cell. This bacterium is unusual among prokaryotes in that sphingolipids comprise a large proportion of its lipids. We here report the presence of 18 molecular species of B. stolpii UKi2 sphingophosphonolipids (SPNLs). (31)P NMR spectroscopy and analysis of P(i) released by a differential hydrolysis protocol confirmed the phosphonyl nature of these lipids. The SPNLs were dominated by those with 1-hydroxy-2-aminoethane phosphonate (hydroxy-aminoethylphosphonate) polar head groups; aminoethylphosphonate was also detected in minor SPNL components. The long-chain bases (LCBs) were dominated by C(17) iso-branched phytosphingosine; C(17) iso branched dihydrosphingosine was also present in some SPNLs. The N-linked fatty acids were predominantly iso-branched and most contained an alpha-hydroxy group (C(15) alpha-hydroxy fatty acid was the major fatty acid). Minor molecular species containing nonhydroxy fatty acids were also detected. The definitive iso structures of the predominant fatty acids and LCBs present in the B. stolpii SPNLs were established using (13)C and (3)H nuclear magnetic resonance spectroscopy; less than 20% were unbranched. Detection and analyses of intact compounds by MS-MS were performed by a hybrid quadrupole time-of-flight (Q-TOF II) MS equipped with an electrospray ionization source. Analyses of peracetylated derivatives verified the structural assignments of these lipids. PMID- 17123829 TI - Sequence specificity and efficiency of protein N-terminal methionine elimination in wheat-embryo cell-free system. AB - Recent improvements in wheat-embryo cell-free translation resulted in a highly productive system for protein preparation. To clarify N-terminal processing of the cell-free system in a preparative-scale (> mg protein product per ml), 20 mutant variants of maltose-binding protein (MalE), each having a different penultimate residue in the sequence Met-Xaa-Ile-Glu-, and 20 glutathione S transferase (GST) variants, having Met-Xaa-Pro-Ile-sequence, were designed and synthesized. The MalE and GST proteins were purified by amylose-resin and glutathione columns, respectively, followed by analysis of their N-terminal sequences. These investigations revealed that sequence specificity and efficiency of the N-terminal Met (N-Met) elimination in the cell-free system are similar to those reported from investigations in cellular systems or in the wheat-embryo cell-free protein expression system in analytical scale (approximately 10 microg protein product per ml). Cleavage of the N-Met is basically determined by the penultimate amino acid in the polypeptide sequence. In the case of MalE, the cleavage was efficient when the penultimate residue was Ala, Cys, Gly, Pro, Ser or Thr. But, in the case of GST with Pro as the antepenultimate residue, the efficiency was significantly reduced when the penultimate residue was Gly or Thr. We also confirmed that substitution of the antepenultimate residue in MalE to Pro drastically reduced the efficiency of N-Met cleavage when the penultimate residue was Ala, Gly, Pro, Ser or Thr, indicating inhibitory effects of antepenultimate residue Pro on N-Met elimination. These results clarified sequence-specific functions of the endogenous N-terminal processing machinery in the scaled-up wheat-embryo cell-free translation system. PMID- 17123830 TI - Protective immunity in grouper (Epinephelus coioides) following exposure to or injection with Cryptocaryon irritans. AB - The protective immunity of grouper (Epinephelus coioides) against Cryptocaryon irritans was determined after immunisation by surface exposure or intraperitoneal injection. Specific antibody titres of immunised fish serum and skin culture supernatant were determined by enzyme-linked immunosorbent assay and immobilisation assays. Specific antibody can be detected in some immunised fish at Week 1 and in all immunised fish at Week 2, and the peaks were between Weeks 4 6. Specific antibody was still evident in the serum and skin of immunised fish at Week 8, and provided good protection against challenge with C. irritans. These findings indicated that humoral and skin mucosal immunity play important roles in fish against C. irritans infection. PMID- 17123831 TI - Non-lethal heat shock protects gnotobiotic Artemia franciscana larvae against virulent Vibrios. AB - Brine shrimp Artemia were exposed under gnotobiotic conditions to a non-lethal heat shock (NLHS) from 28 to 32, 37 and 40 degrees C. Different recovery periods (2, 6, 12 and 24h) and different heat-exposure times (15, 30, 45 and 60 min) were tested. After these NLHS, Artemia was subsequently challenged with Vibrio. Challenge tests were performed in stressed and unstressed nauplii at concentrations of 10(7) cells ml(-1) of pathogenic bacteria, Vibrio campbellii and Vibrio proteolyticus. A NLHS with an optimal treatment of 37 degrees C for 30 min and a subsequent 6h recovery period resulted in a cross-protection against pathogenic Vibrio. A 100% increase in the larval survival (P < 0.05) was observed. We have also demonstrated by Western blot that a NLHS increases the expression of HSP-70 in heat-shocked (HS) treated animals. This report is the first to reveal a cross protection of a NLHS against deleterious bacterial challenges in living crustaceans. The putative role of heat shock proteins (HSPs) in this process is discussed. PMID- 17123832 TI - Cloning and expression analysis of goldfish (Carassius auratus L.) prominin. AB - The integral membrane protein known as prominin was first identified on the apical surface of mouse neural epithelial cells as well as on the surface of human haematopoietic progenitor cells. This report describes a prominin-like sequence and expression analysis of the prominin in the goldfish. The predicted amino acid sequence for goldfish prominin shares all of the hallmark structural characteristics of the prominin family, however the relatedness assessed using the percent amino acid identity indicated that goldfish prominin cannot be placed into the current mammalian dichotomy of type 1 or 2. The real time PCR analyses indicated that prominin was broadly expressed in different tissues with particularly high levels observed in the kidney and gill of the goldfish. Goldfish prominin was also found to be differentially expressed in subpopulations of in vitro-derived goldfish macrophages, with the highest expression observed in progenitor cells. PMID- 17123833 TI - Muscular activity during uphill cycling: effect of slope, posture, hand grip position and constrained bicycle lateral sways. AB - Despite the wide use of surface electromyography (EMG) to study pedalling movement, there is a paucity of data concerning the muscular activity during uphill cycling, notably in standing posture. The aim of this study was to investigate the muscular activity of eight lower limb muscles and four upper limb muscles across various laboratory pedalling exercises which simulated uphill cycling conditions. Ten trained cyclists rode at 80% of their maximal aerobic power on an inclined motorised treadmill (4%, 7% and 10%) with using two pedalling postures (seated and standing). Two additional rides were made in standing at 4% slope to test the effect of the change of the hand grip position (from brake levers to the drops of the handlebar), and the influence of the lateral sways of the bicycle. For this last goal, the bicycle was fixed on a stationary ergometer to prevent the lean of the bicycle side-to-side. EMG was recorded from M. gluteus maximus (GM), M. vastus medialis (VM), M. rectus femoris (RF), M. biceps femoris (BF), M. semimembranosus (SM), M. gastrocnemius medialis (GAS), M. soleus (SOL), M. tibialis anterior (TA), M. biceps brachii (BB), M. triceps brachii (TB), M. rectus abdominis (RA) and M. erector spinae (ES). Unlike the slope, the change of pedalling posture in uphill cycling had a significant effect on the EMG activity, except for the three muscles crossing the ankle's joint (GAS, SOL and TA). Intensity and duration of GM, VM, RF, BF, BB, TA, RA and ES activity were greater in standing while SM activity showed a slight decrease. In standing, global activity of upper limb was higher when the hand grip position was changed from brake level to the drops, but lower when the lateral sways of the bicycle were constrained. These results seem to be related to (1) the increase of the peak pedal force, (2) the change of the hip and knee joint moments, (3) the need to stabilize pelvic in reference with removing the saddle support, and (4) the shift of the mass centre forward. PMID- 17123834 TI - Hippocampal volume is as variable in young as in older adults: implications for the notion of hippocampal atrophy in humans. AB - Previous studies in humans have shown the presence of an age-related reduction of hippocampal (HC) volume, as well as the presence of reduced HC volume in psychiatric populations suffering from schizophrenia, depression or post traumatic stress disorder. Altogether, these data suggested that aging or psychiatric disease can have neurotoxic effects on the hippocampus, and lead to HC atrophy. However, these two sets of findings imply that HC volume in young healthy adults should present less variability than HC volume in older adults and psychiatric populations. In the present study, we assessed HC volume in 177 healthy men and women aged from 18 to 85 years of age. We show that the dispersion around the mean of HC volume is not different in young and older adults, so that 25% of young healthy adults present HC volume as small as the average participants aged 60 to 75 years. This shows that HC volume is as variable in young as in older adults and suggests that smaller HC volume attributed to the aging process in previous studies could in fact represent HC volume determined early in life. We also report that within similar age groups, the percentage of difference in HC volume between the individuals with the smallest HC volume (smallest quartile) and the group average is greater than the percentage of difference reported to exist between psychiatric populations and normal control in recent meta-analyses. Taken together, these results confront the notion of hippocampal atrophy in humans and raise the possibility that pre determined inter-individual differences in HC volume in humans may determine the vulnerability for age-related cognitive impairments or psychopathology throughout the lifetime. PMID- 17123835 TI - Association between cerebral glutamate and human behaviour: the sensation seeking personality trait. AB - INTRODUCTION: Brain imaging studies have linked the anterior cingulate cortex (ACC) to motivation, drive, and personality traits like novelty and sensation seeking. Animal studies have shown glutamatergic neurotransmission to be important in ACC function as well as motivated behaviour. However, the role of glutamate in related personality traits like sensation seeking has not been investigated in humans. METHODS: The associations between sensation seeking personality scores and absolute glutamate concentrations in the ACC and the hippocampal region measured by 3-Tesla proton magnetic resonance spectroscopy (1H MRS) were investigated. RESULTS: ACC glutamate concentration was negatively correlated with the sensation seeking sum score and the experience seeking subscore. A weak negative correlation was also observed between the hippocampal glutamate and the sensation seeking sum score. The reexamination of the glutamate concentration after 4 weeks revealed a similar relationship with sensation seeking. DISCUSSION: Although preliminary, the results are in line with the key role of the ACC for motivation and executive control and with the impact of glutamate on novelty related behaviour observed in animal experiments. The role of the hippocampus for novelty processing is discussed. Glutamate measurement with 1H-MRS may facilitate the understanding of biological underpinnings of personality traits and psychiatric diseases associated with dysfunctions in motivation and drive. PMID- 17123836 TI - Diffusion tensor studies dissociated two fronto-temporal pathways in the human memory system. AB - Recent functional neuroimaging studies have shown that multiple cortical areas are involved in memory encoding and retrieval. However, the underlying anatomical connections among these memory-related areas in humans remain elusive due to methodological limitations. Diffusion tensor imaging (DTI) is a technique based on detecting the diffusion of water molecules from magnetic resonance images. DTI allows non-invasive mapping of anatomical connections and gives a comprehensive picture of connectivity throughout the entire brain. By combining functional magnetic resonance imaging (fMRI) and DTI, we show that memory-related areas in the left dorsolateral prefrontal cortex (DLPFC) and the left ventrolateral prefrontal cortex (VLPFC) each connect with memory-related areas in the left temporal cortex. This result suggests there are two pathways between prefrontal cortex and temporal cortex related to the human memory system. PMID- 17123837 TI - Phylogenetic analysis of honey bee behavioral evolution. AB - DNA sequences from three mitochondrial (rrnL, cox2, nad2) and one nuclear gene (itpr) from all 9 known honey bee species (Apis), a 10th possible species, Apis dorsata binghami, and three outgroup species (Bombus terrestris, Melipona bicolor and Trigona fimbriata) were used to infer Apis phylogenetic relationships using Bayesian analysis. The dwarf honey bees were confirmed as basal, and the giant and cavity-nesting species to be monophyletic. All nodes were strongly supported except that grouping Apis cerana with A. nigrocincta. Two thousand post-burnin trees from the phylogenetic analysis were used in a Bayesian comparative analysis to explore the evolution of dance type, nest structure, comb structure and dance sound within Apis. The ancestral honey bee species was inferred with high support to have nested in the open, and to have more likely than not had a silent vertical waggle dance and a single comb. The common ancestor of the giant and cavity-dwelling bees is strongly inferred to have had a buzzing vertical directional dance. All pairwise combinations of characters showed strong association, but the multiple comparisons problem reduces the ability to infer associations between states between characters. Nevertheless, a buzzing dance is significantly associated with cavity-nesting, several vertical combs, and dancing vertically, a horizontal dance is significantly associated with a nest with a single comb wrapped around the support, and open nesting with a single pendant comb and a silent waggle dance. PMID- 17123838 TI - Phylogenetic relationships among anchovies, sardines, herrings and their relatives (Clupeiformes), inferred from whole mitogenome sequences. AB - The relationships among and within the main lineages of the order Clupeiformes have been explored in few morphological studies and still remain poorly understood. Using whole mitogenome sequences, we inferred the relationships among 25 clupeiform species, sampled from each clupeiform family and subfamily, and a large selection of non-clupeiform teleosts. Our character sets, including unambiguously aligned, concatenated mitogenome sequences that we have divided into four (1st and 2nd codon positions, tRNA genes, and rRNA genes) or five partitions (same as before plus the transversions at 3rd codon positions, using 'RY' coding), were analyzed by the partitioned Bayesian method. The result strongly supported the monophyly of the Clupeiformes within the Otocephala, with Denticeps clupeoides as the sister group of a clade comprising all the remaining clupeiforms species (= suborder Clupeoidei). Within the Clupeoidei, the family Engraulidae was the sister group of the remaining taxa, comprising members of Sundasalangidae, Pristigasteridae, Clupeidae and Chirocentridae. Relationships among the latter four families remained ambiguous. In particular, the position of the Chirocentridae was difficult to estimate possibly owing to its higher molecular evolutionary rate. Of the five subfamilies in the family Clupeidae, monophylies of three (Alosinae, Clupeinae and Dorosomatinae) were statistically rejected. Instead, our mitogenomic data provide strong support for new clades within the Clupeidae, some of which are composed of members of more than one of the previously accepted subfamilies. PMID- 17123839 TI - Complex biogeographic history of the cuckoo-shrikes and allies (Passeriformes: Campephagidae) revealed by mitochondrial and nuclear sequence data. AB - The Campephagidae (minivets, cuckoo-shrikes and trillers, seven genera and 81 species) represents an Old World corvid clade of tropical birds with a mixed diet that forage in different manners, i.e., flycatching, foliage-gleaning or shriking. This family has never been the focus of any phylogenetic survey, so their phylogenetic relationships and biogeographic history remain largely unknown. To address these questions, we sequenced four loci (ND2, myoglobin intron-2, GAPDH intron-11 and c-mos) for 27 species of Campephagidae. Our analyses suggest that Coracina consists of five unrelated lineages and that Lalage is paraphyletic. Our dating analyses, using a Bayesian relaxed-clock method, indicate that the split between the Indo-Malayan genus Pericrocotus and the remaining Campephagidae occurred synchronously with other splits involving Indo-Malayan corvid lineages, suggesting that several lineages of Indo-Malayan birds were isolated at the same time from their closest relatives. The African stock of cuckoo-shrikes is likely the result of three independent trans-oceanic dispersals from Australasia. PMID- 17123840 TI - A molecular phylogeny of the nightjars (Aves: Caprimulgidae) suggests extensive conservation of primitive morphological traits across multiple lineages. AB - We report a molecular re-assessment of the classification of the nightjars which draws conclusions that are strongly at odds with the traditional, morphology based classifications. We used maximum likelihood and Bayesian methods to compare the cytochrome b gene for 14 species from seven of the 15 genera of the Caprimulgidae and partial cytochrome b sequence data was available for a further seven species including three further genera. We found that within the Caprimulgidae there were four geographically isolated clades with bootstrap support greater than 70%. One of these clades contained just Chordeiles species, the remaining three clades each contained a mixture of genera including Caprimulgus sp. A clade of exclusively South American nightjars included the genera Caprimulgus, Uropsalis, Eleopthreptus and Hydropsalis. A clade of African and Eurasian birds included Caprimulgus and Macrodipteryx. Phalaenoptilus nuttallii and Caprimulgus vociferous formed a clade of North American birds. Two ecological factors appear to make morphological classification potentially misleading: first, the apparent retention of primitive anti-predator and foraging related traits across genetically divergent groups; second, rapid divergence in other traits, especially those related to mating, which generate high levels of morphological divergence between species that are genetically very similar. The cytochrome b data suggests that the genus Caprimulgus is not monophyletic and is restricted to Africa and Eurasia and that Caprimulgus species from outside this area have been misclassified as a consequence of retention of primitive adaptations for crepuscular/nocturnal living. Some other genera also appear to have little support from the cytochrome b data. PMID- 17123841 TI - BCP crystals increase prostacyclin production and upregulate the prostacyclin receptor in OA synovial fibroblasts: potential effects on mPGES1 and MMP-13. AB - OBJECTIVE: To investigate the potential involvement of prostacyclin in basic calcium phosphate (BCP) crystal-induced responses in osteoarthritic synovial fibroblasts (OASF). METHODS: OASF grown in culture were stimulated with BCP crystals. Prostacyclin production was measured by enzyme immunoassay. Expression of messenger RNA (mRNA) transcripts was assessed by real-time polymerase chain reaction (PCR). Expression of prostacyclin synthase (PGIS) and the prostacyclin (IP) receptor was measured. The effects of iloprost, a prostacyclin analogue, on expression of genes implicated in osteoarthritis such as microsomal prostaglandin E2 synthase 1 (mPGES1) and matrix metalloproteinases (MMPs) were also studied. FPT inhibitor II, a farnesyl transferase inhibitor, was used to antagonize iloprost-induced responses. RESULTS: BCP crystal stimulation led to a five-fold increase in prostacyclin production in OASF compared to untreated cells. This induction was attenuated by cyclooxygenase (COX)-2 and COX-1 inhibition at 4 and 32h, respectively. PGIS and IP receptor transcripts were constitutively expressed in OASF. BCP crystals upregulated IP receptor expression two-fold. While iloprost diminished BCP crystal-stimulated IP receptor upregulation, the inhibitory effect of iloprost was blocked by the farnesyl transferase inhibitor. In addition, iloprost upregulated mPGES1 and downregulated MMP-13 expression in BCP crystal stimulated OASF, effects that were not influenced by the farnesyl transferase inhibitor. CONCLUSIONS: These data showed for the first time that BCP crystals can increase prostacyclin production and upregulate expression of the IP receptor in OASF. The potential of prostacyclin to influence BCP crystal-stimulated responses was supported by the effects of iloprost on the expression of the IP receptor, mPGES1 and MMP-13. These data demonstrate the potential involvement of prostacyclin in BCP crystal-associated osteoarthritis (OA) and suggest that inhibition of PG synthesis with non-steroidal anti-inflammatory drugs may have both deleterious and beneficial effects in BCP crystal-associated OA. PMID- 17123842 TI - Cellular and extracellular behavior in the gerbil (Meriones unguiculatus) ventral prostate following different types of castration and the consequences of testosterone replacement. AB - Mongolian gerbils (Meriones unguiculatus) were grouped into two experimental groups: GEx.01 suffered orchiectomy and after 30 days received doses of testosterone cipionate (T), while GEx.02 received weekly and alternated doses of the anti-androgens cyproterone acetate and flutamide for 30 days, and the animals were then euthanized. Structural evaluation reveals a more intense reduction in epithelial height in GEx.02. Smooth muscle cells (SMC) presented a star-shaped aspect after 30 days of hormonal ablation and basal membrane was shown to be more intensely grooved in GEx.01. In both groups, after hormonal replacement, recovery in epithelial height could be noted and the SMC presented its phenotypes, but an increase in RER was seen, characterizing a modulation from its contractile to secreting phenotype. In conclusion, the prostate presented involution capacity after androgen ablation and the ability to reorganize after hormonal replacement, but events resulting from orchiectomy and subsequent T replacement were shown to be more aggressive to the prostate. PMID- 17123843 TI - In vitro effect of biogenic amines on the hormone content of immune cells of the peritoneal fluid and thymus. Is there a hormonal network inside the immune system? AB - Immune cells contain different hormones and hormone-like molecules, such as insulin, endorphin, triiodothyronine (T3) histamine, serotonin. In earlier in vitro experiments insulin down-regulated histamine, serotonin and T3 content of thymus cells. Now we studied the effect of biogenic amines on the endorphin, T3, serotonin and histamine content of rat peritoneal and thymic cells. Cells were obtained from male rats of 100g body weight. 100 ng/ml serotonin or 300 ng/ml histamine was added for 30 min. After that the cells were prepared for flow cytometric analysis with antibodies to endorphin, T3, histamine and serotonin as primary antibodies and anti-rabbit IgG as secondary antibody. Finishing the measurements the cells were also studied by confocal microscopy. T3 concentration (binding of anti-T3 antibody) increased in peritoneal mast cells after serotonin treatment and in the monocyte-macrophage-granulocyte group after histamine treatment. Thymocytes' T3 content radically decreased after both treatments. Serotonin and histamine treatment also radically reduced the amine content of each other. Endorphin level was resistant to hormonal treatments. The results call attention to a possible hormonal network inside the immune system in which hormones produced by the immune cells themselves can influence each other. PMID- 17123844 TI - Cajal bodies and interchromatin granule clusters in cricket oocytes: composition, dynamics and interactions. AB - The organization and molecular composition of complicated Cajal bodies (CBs) and interchromatin granule clusters (IGCs) in oocytes of the house cricket, Acheta domesticus, were studied using immunofluorescent/confocal and Immunogold labeling/electron microscopy. In A. domesticus oocytes, the CB consists of the fibrillar matrix and a central cavity containing a predominantly granular body with insertions of tightly packed fibrillar material. The latter structure was identified as an "internal" IGC, since it is enriched with the SC35 protein, a marker for IGCs. The IGCs located outside the CB were also identified. Microinjections of the fluorescein-tagged U7 snRNA into the ooplasm showed the targeting of the U7 to the matrix of the CB. Some other essential CB components (coilin, snRNPs, fibrillarin) were found to be colocalized in the matrix of the CB. Neither confocal nor Immunogold microscopy revealed significant amounts of RNA polymerase II (pol II) in the CB of A. domesticus oocytes. The splicing factor SC35 was detected in the matrix of the CB. In oocytes treated with DRB, the amount of IGCs in the nucleoplasm increased significantly, granular and fibrillar components of IGCs were segregated, and the fibrillar areas accumulated pol II. Additionally, IG-like granules were shown to display on the surface of the CB probably due to a shifting from the internal IGC. We believe that in A. domesticus oocytes, CBs are involved in nuclear distribution of splicing factors, but their role in pol II transport is less significant. We also suggest that the formation of complicated CBs is a result of interconnection between two different nuclear domains, CBs and IGCs. PMID- 17123845 TI - Tissue inhibitor of metalloproteinase-2 (TIMP-2) location in the ventral, lateral, dorsal and anterior lobes of rat prostate by immunohistochemistry. AB - Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play a major role in extracellular matrix component degradation in several normal and abnormal tissue situations; they are also found in human seminal plasma. MMPs have been found in rat prostate secretions and are nearly lobe specific in expression pattern. The aim of this study was to evaluate whether TIMP-2, like other semen components, is expressed differently from different rat prostatic lobes. Immunohistochemical staining was performed in both young and adult rat ventral (VP), lateral (LP), dorsal (DP), and anterior (AP) prostatic lobes and confirmed by western blotting. TIMP-2 expression was found in the epithelial cells in the following sequence: LP>AP>DP>VP, in both young and adult rats. In this study, 100% of adult LP presented histological signs of prostatitis, where TIMP-2 immunostaining was positive in normal epithelium even with intraluminal neutrophils, but was reduced or absent in the epithelium with intraepithelial leukocytes or with periductal stroma disorganization associated with mononuclear cell infiltration. However, TIMP-2 expression in LP was not induced by prostatitis, since younger rat LPs were also strongly TIMP-2 positive. The distal and intermediate VP regions were TIMP-2 negative, but the proximal regions were strongly stained. Western blotting results confirmed the high TIMP-2 expression in the LP lobe. Thus, TIMP-2 is expressed differently between the prostatic lobes and is another nearly lobe-specific protein, which plays a role in the regulation of MMP activity in seminal plasma and glandular homeostasis. TIMP-2 is also another regional ductal variation of VP. Further studies should address whether TIMP-2 expression is related to the highest incidence of rat LP prostatitis and adenocarcinoma. PMID- 17123846 TI - SPACE and EVA-3S trials: the need of standards for carotid stenting. PMID- 17123847 TI - The role of sodium channels in neuropathic pain. AB - Our knowledge of the ion channels, receptors and signalling mechanisms involved in pain pathophysiology, and which specific channels play a role in subtypes of pain such as neuropathic and inflammatory pain, has expanded considerably in recent years. It is now clear that in the neuropathic state the expression of certain channels is modified, and that these changes underlie the plasticity of responses that occur to generate inappropriate pain signals from normally trivial inputs. Pain is modulated by a subset of the voltage-gated sodium channels, including Nav1.3, Nav1.7, Nav1.8 and Nav1.9. These isoforms display unique expression patterns within specific tissues, and are either up- or down-regulated upon injury to the nervous system. Here we describe our current understanding of the roles of sodium channels in pain and nociceptive information processing, with a particular emphasis on neuropathic pain and drugs useful for the treatment of neuropathic pain that act through mechanisms involving block of sodium channels. One of the future challenges in the development of novel sodium channel blockers is to design and synthesise isoform-selective channel inhibitors. This should provide substantial benefits over existing pain treatments. PMID- 17123848 TI - The effects of ruthenium tetraammine compounds on vascular smooth muscle. AB - The time course of the relaxation effect induced by a single dose (3 x 10(-6) mol/L) of trans-[Ru(NH3)4L(NO)]3+ (L=nic, 4-pic, py, imN, P(OEt)3, SO(3)(2-), NH3, and pz) species and sodium nitroprusside (4 x 10(-9) mol/L) was studied in aortic rings without endothelium and pre-contracted with noradrenaline (1 x 10( 6) mol/L). All the compounds induced a relaxing effect in the aortic rings, but the intensity and time of relaxation were different. Only the species where L=py, 4-pic, and P(OEt)3 were able to induce 100% (99-100%) of the relaxing effect during the assay. trans-[Ru(NH3)4(L)(NO)]3+ (L=SO(3)(2-) and NH3) showed the lowest relaxing effect (36 and 37%, respectively) when compared with the other compounds. Relationship was observed between the time corresponding to half of the maximum relaxation intensity observed and, respectively, k-NO, E0'[Ru(NO)]3+/[Ru(NO)]2+ in trans-[Ru(NH3)4(L)(NO)]3+ species and E0'Ru(III)/Ru(II) in trans-[Ru(NH3)4(L)(H2O)]3+ ions. These relationships strongly suggested that the NO liberation from the reduced nitrosyl complexes was responsible for the observed relaxation. PMID- 17123849 TI - High-resolution NMR with resistive and hybrid magnets: deconvolution using a field-fluctuation signal. AB - A method for compensating effect of field fluctuation is examined to attain high resolution NMR spectra with resistive and hybrid magnets. In this method, time dependence of electromotive force induced for a pickup coil attached near a sample is measured synchronously with acquisition of NMR. Observed voltage across the pickup coil is converted to field fluctuation data, which is used to deconvolute NMR signals. The feasibility of the method is studied by (79)Br MAS NMR of KBr under a 30T magnetic field of a hybrid magnet. Twenty single-scan NMR signals were accumulated after the manipulation, resulting in a high-resolution NMR spectrum. PMID- 17123850 TI - A unilateral NMR magnet for sub-structure analysis in the built environment: the Surface GARField. AB - A new, portable NMR magnet with a tailored magnetic field profile and a complementary radio frequency sensor have been designed and constructed for the purpose of probing in situ the sub-surface porosity of cement based materials in the built environment. The magnet is a one sided device akin to a large NMR-MOUSE with the additional design specification of planes of constant field strength /B0/ parallel to the surface. There is a strong gradient G in the field strength perpendicular to these planes. As with earlier GARField magnets, the ratio G//:B0/ is a system constant although the method of achieving this condition is substantially different. The new magnet as constructed is able to detect signals 50mm (1H NMR at 3.2 MHz) away from the surface of the magnet and can profile the surface layers of large samples to a depth of 35-40 mm by moving the magnet, and hence the resonant plane of the polarising field, relative to the sample surface. The matching radio frequency excitation/detector coil has been designed to complement the static magnetic field such that the polarising B0 and sensing B1 fields are, in principal, everywhere orthogonal. Preliminary spatially resolved measurements are presented of cement based materials, including two-dimensional T1-T2 relaxation correlation spectra. PMID- 17123851 TI - Dynamics of paramagnetic agents by off-resonance rotating frame technique in the presence of magnetization transfer effect. AB - The simple method for measuring the rotational correlation time of paramagnetic ion chelates via off-resonance rotating frame technique is challenged in vivo by the magnetization transfer effect. A theoretical model for the spin relaxation of water protons in the presence of paramagnetic ion chelates and magnetization transfer effect is described. This model considers the competitive relaxations of water protons by the paramagnetic relaxation pathway and the magnetization transfer pathway. The influence of magnetization transfer on the total residual z magnetization has been quantitatively evaluated in the context of the magnetization map and various difference magnetization profiles for the macromolecule conjugated Gd-DTPA in cross-linked protein gels. The numerical simulations and experimental validations confirm that the rotational correlation time for the paramagnetic ion chelates can be measured even in the presence of strong magnetization transfer. This spin relaxation model also provides novel approaches to enhance the detection sensitivity for paramagnetic labeling by suppressing the spin relaxations caused by the magnetization transfer. The inclusion of the magnetization transfer effect allows us to use the magnetization map as a simulation tool to design efficient paramagnetic labeling targeting at specific tissues, to design experiments running at low RF power depositions, and to optimize the sensitivity for detecting paramagnetic labeling. Thus, the presented method will be a very useful tool for the in vivo applications such as molecular imaging via paramagnetic labeling. PMID- 17123852 TI - Association between plasma IL-6, the IL6 -174G>C gene variant and the metabolic syndrome in type 2 diabetes mellitus. AB - Elevated plasma interleukin-6 (IL-6) is associated with coronary heart disease (CHD), impaired glucose tolerance (IGT), and type 2 diabetes (T2DM). We and others have described an association between the human interleukin-6 -174G>C gene variant and body mass index (BMI). Within our previous sample of subjects with T2DM, we measured plasma IL-6 and grouped subjects by the WHO-defined metabolic syndrome, in order to study the association between the -174G>C gene variant, plasma IL-6 and the metabolic syndrome (and component parts). Genotype was obtained in 571 Caucasian subjects with plasma IL-6 measures. There was a significant association between genotype and plasma IL-6 (GG vs GC vs CC: 3.23+/ 0.93 pg/ml vs 3.42+/-0.95 pg/ml vs 4.16+/-1.18 pg/ml, p=0.02; for GG/GC vs CC p=0.008). No interactions were observed between genotype and the individual components of the metabolic syndrome in determining plasma IL-6. Increased plasma IL-6 was also associated with the number of components (none vs 1 vs 2 vs > or =3: 2.67+/-0.71 pg/ml vs 2.97+/-0.94 pg/ml vs 4.07+/-1.13 pg/ml, p<0.0001). Within the sample, 76% of CC compared to 56% of GG subjects had the metabolic syndrome (p=0.007). Further analysis of association between the genotype and the components of the metabolic syndrome revealed no further associations than that with BMI previously described. The association of this gene variant with the metabolic syndrome is intimately linked with obesity per se. Further prospective work is required to explore the effect of this gene variant in relation to obesity, the metabolic syndrome and 'prediabetes'. PMID- 17123853 TI - [Proteomics and breast cancer]. AB - Breast cancer is the first cause of death between 35 and 55 years. Genetic alterations and modifications in gene expression are found during different steps of tumor progression. These changes are translated at the protein level where quantitative and qualitative modifications are found in tumor compared to normal samples. Similarly to studies aimed at deciphering transcriptional changes important in cancer, proteomic approaches allow the global and comparative study of proteins in normal and pathological samples. The objective of this article is to present common proteomic methods and to review the first published results concerning proteomics studies applied to breast cancer with an emphasis on reports obtained using the SELDI-TOF MS (Surface Enhanced Laser Desorption Ionization Time-Of-Flight Mass Spectrometry). In breast cancer, it is possible to explore the tumoral proteome and/or the blood derived proteome. The first studies are aimed at globally understanding the disease while the latter are aimed at discovering serum proteins or biomarkers useful for the early detection, diagnosis, prognosis and management of cancer. Promising results are obtained using these emerging methods and these novel biomarkers should be validated in the future and will have an important impact for the management of breast cancer patients. PMID- 17123854 TI - p-Aminophenol degradation by ozonation combined with sonolysis: operating conditions influence and mechanism. AB - The degradation of p-aminophenol (PAP) in aqueous solution by sonolysis, by ozonation, and by a combination of both was investigated in laboratory-scale experiments. Operation parameters such as pH, temperature, ultrasonic energy density and ozone dose were optimized with regard to the efficiency of PAP removal. The concentration of PAP during the reaction was detected by high pressure liquid chromatography. The concentrations of ammonium ions and nitrate ions were monitored during the degradation. Intermediate products such as 4 iminocyclohexa-2,5-dien-1-one, phenol, but-2-enedioic acid, and acetic acid were detected by gas chromatography coupled with mass spectrometry. The degradation rate of PAP was higher in the combined system than in the linear combination of separate experiments. The degradation efficiency was decreased rapidly when n butanol was added to the combined reaction system, which showed that some radical reaction might proceed during the laboratory experiments. PMID- 17123855 TI - Sonophotocatalytic degradation of congo red and methyl orange in the presence of TiO2 as a catalyst. AB - A comparative study between the photocatalytic and sonophotocatalytic oxidation process of congo red was carried out using titanium dioxide as a catalyst. The effect of a number of parameters, such as the initial concentration of dye, the presence of oxygen and ultrasound, the TiO(2) crystalline structure and the amount of TiO(2), was studied using an inexpensive reactor. In the second part of this document, the oxidation and reduction processes of methyl orange was studied using the same reactor, but by changing the chemical environment in order to drive either the oxidation or the reduction reaction. The results showed, for all the cases studied, a sinergistic effect between sonolysis and photocatalysis when an ultrasonic bath of 80W was used as a source of ultrasound. On the other hand, the electrochemical experiments showed that the current densities for the oxidation process of the azodyes were higher when a conducting glass electrode covered with a thin film of TiO(2) was used as a working electrode instead of a platinum electrode, showing that the TiO(2) electrode is not passivated by the oxidation reaction. PMID- 17123856 TI - Design of a flotation cell equipped with ultrasound transducers to enhance coal flotation. AB - Ultrasonic treatment is widely used for surface cleaning during physical, chemical and physico-chemical processes in mineral processing. Several research papers and a few industrial applications about the subject suggest that the mechanism behind the positive effect of ultrasound for mineral processing and especially flotation is due to formation of cavitation by ultrasonic energy. Within this study, coal floatability is investigated by use of a specially designed flotation cell equipped with ultrasound transducers with different power, frequency and geometry. The results indicate that ultrasonic treatment during coal flotation positively affects the quality and quantity of the properties of floated coals while using of lesser amounts of reagent than a conventional flotation system. PMID- 17123857 TI - A novel micro tensile tester with feed-back control for viscoelastic analysis of single isolated smooth muscle cells. AB - A novel micro tensile tester was developed to measure the viscoelastic properties of single isolated cells. A specimen cell was held with two glass micropipettes, the tips of which were coated with a urethane resin adhesive. One pipette was rigid and fixed in position, and the other was deflectable and could be moved by a laboratory-made piezoelectric actuator to stretch the specimen. The actuator was made of an originally designed lever mechanism which can magnify the deformation of conventional piezoceramic actuator and allowed for sufficient displacement up to 150 microm. The distance between the two pipette tips was obtained by real-time image processing and kept constant following stepwise stretching with an accuracy of +/-0.1 microm by changing the voltage applied to the piezoceramics under feed-back control. The force applied to the cell was measured by the deflection of a cantilever part of the deflectable pipette. The stress relaxation test was performed for cultured rat aortic smooth muscle cells. The cells were stretched by 75% and their length was kept constant. Viscoelastic analysis with a standard linear solid showed that the relaxation time constant of the isolated smooth muscle cells was 250+/-42 s (n=8, mean+/-S.E.M.) which was over six times longer than that reported for vascular endothelial cells and fibroblasts. PMID- 17123858 TI - Adaptive regularization network based neural modeling paradigm for nonlinear adaptive estimation of cerebral evoked potentials. AB - In this paper we report an adaptive regularization network (ARN) approach to realizing fast blind separation of cerebral evoked potentials (EPs) from background electroencephalogram (EEG) activity with no need to make any explicit assumption on the statistical (or deterministic) signal model. The ARNs are proposed to construct nonlinear EEG and EP signal models. A novel adaptive regularization training (ART) algorithm is proposed to improve the generalization performance of the ARN. Two adaptive neural modeling methods based on the ARN are developed and their implementation and performance analysis are also presented. The computer experiments using simulated and measured visual evoked potential (VEP) data have shown that the proposed ARN modeling paradigm yields computationally efficient and more accurate VEP signal estimation owing to its intrinsic model-free and nonlinear processing characteristics. PMID- 17123859 TI - Design of a knee rotatory kinaesthetic device. AB - The anterior cruciate ligament (ACL) constrains rotatory motion at the knee and is commonly injured during rotational movements in athletic activity. This densely innervated ligament is assumed to play a role in knee proprioception, however, no study has measured proprioception in a manner relevant to either the kinematics of the ligament or the mechanism of injury, partly because of a lack of suitable equipment. The aims of this technical note are to document the development of a novel knee rotatory kinaesthetic device, and to present details of its construction, reproducibility, accuracy and application. The purpose-built device allows rotational movements at the knee to occur with minimal frictional resistance and provides accurate limits to the magnitude of these movements. This allows analysis of subjects' ability to discriminate between movements of differing magnitudes and thus allows calculation of subjects' sensitivity to small differences in magnitude of active knee rotation. Measurements taken with the device had a high level of agreement with those of a calibrated digital inclinometer (ICC=0.99; 95% CI 0.88 to 0.99) with a mean error of 0.24 degrees . The device also demonstrated excellent reproducibility (Pearson's r=1.0). A single case study is presented to detail the clinical application of the device. This novel device allows subjects to perform active knee rotational movements in a closed kinetic chain with discrete, self-paced movement, enabling calculation of movement discrimination. The device is compact and portable enabling testing to be undertaken in remote settings enhancing its clinical applicability. PMID- 17123860 TI - Caveolin-3 and eNOS colocalize and interact in ciliated airway epithelial cells in the rat. AB - In ciliated airway epithelial cells endothelial nitric oxide synthase as well as several other membrane bound proteins are located in the apical cell pole. To date, mechanisms that serve to target and to keep these proteins in this region are unknown. Endothelial nitric oxide synthase is known to target to caveolae by interaction with caveolin-1 or caveolin-3. Since caveolin-1 is found only in a subpopulation of ciliated cells at the basolateral cell membrane, we examined if caveolin-3 could be responsible for the apical localization of endothelial nitric oxide synthase in ciliated cells. We used real-time RT-PCR, laser-assisted microdissection, Western blotting and double-labeling immunohistochemistry to examine the presence of caveolin-3 in the airway epithelium of the rat. Indeed, we found caveolin-3-mRNA as well as protein in ciliated cells throughout the trachea and the bronchial tree. Caveolin-3-immunoreactivity was confined to the apical region and was colocalized with endothelial nitric oxide synthase and the high affinity choline transporter in a compartment distinct from the plasma membrane at the light microscopic level. No caveolae were found in the apical plasma membrane of ciliated cells but a tubulovesicular network was present in the apical region that reached up to the basal bodies of the cilia and was in close contact with mitochondria. Co-immunoprecipitation of caveolin-3 with endothelial nitric oxide synthase verified that both proteins interact in airway ciliated cells. These findings indicate that caveolin-3 is responsible to keep endothelial nitric oxide synthase in a membrane compartment in the apical region of ciliated cells. PMID- 17123861 TI - Follow-up testing of rodent carcinogens not positive in the standard genotoxicity testing battery: IWGT workgroup report. AB - At the Plymouth Third International Workshop on Genotoxicity Testing in June 2002, a new expert group started a working process to provide guidance on a common strategy for genotoxicity testing beyond the current standard battery. The group identified amongst others "Follow-up testing of tumorigenic agents not positive in the standard genotoxicity test battery" as one subject for further consideration [L. Muller, D. Blakey, K.L. Dearfield, S. Galloway, P. Guzzie, M. Hayashi, P. Kasper, D. Kirkland, J.T. MacGregor, J.M. Parry, L. Schechtman, A. Smith, N. Tanaka, D. Tweats, H. Yamasaki, Strategy for genotoxicity testing and stratification of genotoxicity test results-report on initial activities of the IWGT Expert Group, Mutat. Res. 540 (2003) 177-181]. A workgroup devoted to this topic was formed and met on September 9-10, 2005, in San Francisco. This workgroup was devoted to the discussion of when it would be appropriate to conduct additional genetic toxicology studies, as well as what type of studies, if the initial standard battery of tests was negative, but tumor formation was observed in the rodent carcinogenicity assessment. The important role of the standard genetic toxicology testing to determine the mode of action (MOA) for carcinogenesis (genotoxic versus non-genotoxic) was discussed, but the limitations of the standard testing were also reviewed. The workgroup also acknowledged that the entire toxicological profile (e.g. structure-activity relationships, the nature of the tumor finding and metabolic profiles) of a compound needed to be taken into consideration before the conduct of any additional testing. As part of the meeting, case studies were discussed to understand the practical application of additional testing as well as to form a decision tree. Finally, suitable additional genetic toxicology assays to help determine the carcinogenic MOA or establish a weight of evidence (WOE) argument were discussed and formulated into a decision tree. PMID- 17123862 TI - Inhibition of transcellular tumor cell migration and metastasis by novel carba derivatives of cyclic phosphatidic acid. AB - Cyclic phosphatidic acid (1-acyl-sn-glycerol-2,3-cyclic phosphate; cPA) is a naturally occurring analog of lysophosphatidic acid (LPA) with a variety of distinctly different biological activities from those of LPA. In contrast to LPA, a potent inducer of tumor cell invasion, palmitoyl-cPA inhibits FBS- and LPA induced transcellular migration and metastasis. To prevent the conversion of cPA to LPA we synthesized cPA derivatives by stabilizing the cyclic phosphate ring; to prevent the cleavage of the fatty acid we generated alkyl ether analogs of cPA. Both sets of compounds were tested for inhibitory activity on transcellular tumor cell migration. Carba derivatives, in which the phosphate oxygen was replaced with a methylene group at either the sn-2 or the sn-3 position, showed much more potent inhibitory effects on MM1 tumor cell transcellular migration and the pulmonary metastasis of B16-F0 melanoma than the natural pal-cPA. The antimetastatic effect of carba-cPA was accompanied by the inhibition of RhoA activation and was not due to inhibition of the activation of LPA receptors. PMID- 17123863 TI - Diastolic dysfunction and autonomic abnormalities in patients with systolic heart failure. AB - BACKGROUND: Patients with systolic heart failure (SHF) often have concomitant diastolic dysfunction (DD). SHF is associated with decreased heart rate variability (HRV), but the impact of degree of DD on HRV in SHF is unclear. METHODS AND RESULTS: HRV was measured in 139 patients, aged 64+/-12 years, 74% male, LVEF 30+/-8%. Patients had stable NYHA class II-III CHF on ACE inhibitors or ATII receptor blockers, with LVEFor=200 pg/ml. Subjects underwent 2-D echocardiography with Doppler assessment and 24-h Holters. Patients were categorized as having impaired relaxation (E-deceleration time>2 SD above age-adjusted normal values (AANV), E/Aor=1; N=30), pseudonormal (E-deceleration time within 2 SD of AANV, E/A=1-2, systolic/diastolic pulmonary vein flow<1; N=25) or restrictive filling patterns (E-deceleration time>2 SD below AANV or/and E/A ratio>or=2; N=84) Differences were adjusted for clinical covariates using UNIANOVA, p<0.05. HRV was reduced and BNP higher in pseudonormal patients compared to impaired relaxation, but this difference was only significant for restrictive vs. impaired filling. Differences remained significant after adjustment for covariates. CONCLUSION: Significantly more abnormal HRV, reflecting greater cardiac autonomic dysfunction, is associated with restrictive DD compared to impaired relaxation. PMID- 17123864 TI - The expression of a plant-type ferredoxin redox system provides molecular evidence for a plastid in the early dinoflagellate Perkinsus marinus. AB - Perkinsus marinus is a parasitic protozoan with a phylogenetic positioning between Apicomplexa and dinoflagellates. It is thus of interest for reconstructing the early evolution of eukaryotes, especially with regard to the acquisition of secondary plastids in these organisms. It is also an important pathogen of oysters, and the definition of parasite-specific metabolic pathways would be beneficial for the identification of efficient treatments for infected mollusks. Although these different scientific interests have resulted in the start of a genome project for this organism, it is still unknown whether P. marinus contains a plastid or plastid-like organelle like the related dinoflagellates and Apicomplexa. Here, we show that in vitro-cultivated parasites contain transcripts of the plant-type ferredoxin and its associated reductase. Both proteins are nuclear-encoded and possess N-terminal targeting sequences similar to those characterized in dinoflagellates. Since this redox pair is exclusively found in cyanobacteria and plastid-harboring organisms its presence also in P. marinus is highly indicative of a plastid. We also provide additional evidence for such an organelle by demonstrating pharmacological sensitivity to inhibitors of plastid-localized enzymes involved in fatty acid biosynthesis (e.g. acetyl-CoA carboxylase) and by detection of genes for three enzymes of plastid localized isoprenoid biosynthesis (1-deoxy-D-xylulose 5-phosphate reductoisomerase, (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate reductase, and (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate synthase). PMID- 17123865 TI - Computational identification of microRNAs and their targets. AB - MicroRNAs (miRNAs) are one class of newly identified riboregulators of gene expression in many eukaryotic organisms. They play important roles in multiple biological and metabolic processes, including developmental timing, signal transduction, cell maintenance and differentiation, diseases and cancers. miRNAs regulate gene expression at the posttranscriptional level by directly cleaving targeted mRNAs or repressing translation. Although the founding members of miRNAs were discovered by genetic screening approaches, experimental approaches were limited by their low efficiency, time consuming, and high cost. As an alternative, computational approaches were developed. Computational approaches for identifying miRNAs are based on the following major characteristics of miRNAs: hairpin-shaped secondary structures, high conservation for some miRNAs, and high minimal folding free energy index (MFEI). Computational approaches also play an important role in identifying miRNA targets. A majority of known miRNAs and their targets were identified by computational approaches. Several web-based or non-web-based computer software programs are publicly available for predicting miRNAs and their targets. PMID- 17123866 TI - People with epilepsy are often perceived as violent. AB - This study explores the possibility that people with epilepsy are commonly and persistently perceived as potentially violent during and between seizures. In 1981 and again in 2006, we assessed responses to a questionnaire that includes vignettes and direct questions about violence in epilepsy. Groups sampled were medical and law students, physicians, child care workers, the general public, and people with epilepsy (n=271 in 1981 and n=388 in 2006). Nearly half of the respondents believed that violence was possible or likely during a seizure. Almost all groups answered at least 40% of questions incorrectly; the exception was physicians, who answered 20% incorrectly. Responses were fairly stable over the 25-year interval. Logistic regression revealed few predictors: older responders and physicians had more correct answers, whereas law students had poorer performance. These results indicate that fear of violence at the hands of people with epilepsy is prevalent and may contribute to stigma. PMID- 17123867 TI - A unicenter, randomized, double-blind, parallel-group, placebo-controlled study of Melatonin as an Adjunct in patients with acute myocaRdial Infarction undergoing primary Angioplasty The Melatonin Adjunct in the acute myocaRdial Infarction treated with Angioplasty (MARIA) trial: study design and rationale. AB - BACKGROUND: Experimental studies have documented the beneficial effects of the endogenously produced antioxidant, melatonin, in reducing tissue damage and limiting cardiac pathophysiology in models of experimental ischemia-reperfusion. Melatonin confers cardioprotection against ischemia-reperfusion injury most likely through its direct free radical scavenging activities and its indirect actions in stimulating antioxidant enzymes. These actions of melatonin permit it to reduce molecular damage and limit infarct size in experimental models of transient ischemia and subsequent reperfusion. STUDY DESIGN: The Melatonin Adjunct in the acute myocaRdial Infarction treated with Angioplasty (MARIA) trial is an unicenter, prospective, randomized, double-blind, placebo-controlled, phase 2 study of the intravenous administration of melatonin. The primary efficacy end point of this study is to determine whether melatonin treatment reduces infarct size determined by the cumulative release of alpha-hydroxybutyrate dehydrogenase (area under the curve: 0 to 72 h). Other secondary end points will be the clinical events occurring within the first 90 days: death, sustained ventricular arrhythmias, resuscitation from cardiac arrest, cardiogenic shock, heart failure, major bleedings, stroke, need for revascularization, recurrent ischemia, re infarctions and rehospitalization. IMPLICATIONS: The MARIA trial tests a novel pharmacologic agent, melatonin, in patients with acute myocardial infarction and the hypothesis that it will confer cardioprotection against ischemia-reperfusion injury. If successful, the finding would support the use of melatonin in therapy of ischemic-reperfusion injury of the heart. PMID- 17123868 TI - Presentation in relation to publication of results from clinical trials. AB - BACKGROUND: Results from clinical trials are typically disseminated first by presentation at scientific meetings. An important question has to do with the role of presentation in improving the quality of manuscripts submitted to the journals as well as the effect of presentation in speeding, or delaying subsequent publication. The aim of this research is focused on presentation practices of trialists to examine their effect on the timing of publications of clinical trial results. METHODS: Six hundred and one (601) trials published in 1996 and 1997 were identified via MEDLINE using medical subject heading "clinical trials" or the occurrence of the term in the text and by limiting to publication type "clinical trial". Authors of those trials were surveyed to determine prior presentation history for the identified trials. RESULTS: Among the 601 trials identified, complete responses to questionnaires were obtained for 379 (63%) trials. The median time from completion to first submission of the primary results manuscript was 11 months and the median time from completion to publication was 25 months for the 220 trials involving presentation prior to submission for publication. The corresponding median times from completion to first submission and publication for the subset of trials not involving presentation prior to the submission were 8 and 19 months (159 trials), respectively. The adjusted relative hazard for publication for trials involving presentation prior to first submission was 0.55 versus trials not involving presentation prior to first submission (95% confidence interval, 0.44 to 0.69). CONCLUSION: Despite the importance of dissemination of results prior to publication, investigators should carefully weigh a potential gain in quality against a potential for delay in submission of the primary results manuscript by presentation at scientific meetings. The findings of our study suggest that presentation prior to submission may increase time to publication. Inclusion of presentation dates in clinical trial registers should be considered to allow future studies investigating presentation and publication practices. PMID- 17123869 TI - Spatial and temporal expression of POF1B, a gene expressed in epithelia. AB - Mammalian epithelia possess specialized cellular components that provide an impermeable barrier between two different environments. In particular, in the skin, mitotically dividing cells undergo a programmed set of morphological and biochemical changes leading to the establishment of the epidermal permeability barrier (EPB) to prevent escape of moisture and entrance of toxic molecules. Many different skin proteins are involved in the process but not all have been identified. We report here the results of the expression studies of a novel gene, highly and specifically expressed in the granular layer of the epidermis and in the epithelia of the oro-pharyngeal and gastro-intestinal tracts. Our data show that during mouse development Pof1b expression is activated in the external layers of the epidermis just prior to formation of the EPB. PMID- 17123870 TI - High electrical field effects on cell membranes. AB - Electrical charging of lipid membranes causes electroporation with sharp membrane conductance increases. Several recent observations, especially at very high field strength, are not compatible with the simple electroporation picture. Here we present several relevant experiments on cell electrical responses to very high external voltages. We hypothesize that, not only are aqueous pores created within the lipid membranes, but that nanoscale membrane fragmentation occurs, possibly with micelle formation. This effect would produce conductivity increases beyond simple electroporation and display a relatively fast turn-off with external voltage. In addition, material loss can be expected at the anode side of cells, in agreement with published experimental reports at high fields. Our hypothesis is qualitatively supported by molecular dynamics simulations. Finally, such cellular responses might temporarily inactivate voltage-gated and ion-pump activity, while not necessarily causing cell death. This hypothesis also supports observations on electrofusion. PMID- 17123871 TI - Impaired platelet mitochondrial activity in Alzheimer's disease and mild cognitive impairment. AB - Mitochondrial abnormalities are found in Alzheimer's disease (AD), but previous reports have not examined at-risk groups. In subjects with AD, mild cognitive impairment (MCI), and non-demented aged controls, platelet and lymphocyte mitochondria were isolated and analyzed for Complexes I, III, and IV of the electron transport chain. Western blots were used to control for differential enrichment of samples. Results demonstrated significant declines in Complexes III and IV in AD, and a significant decline in Complex IV in MCI. This report confirms mitochondrial deficiencies in AD, extends them to MCI, and suggests they are present at the earliest symptomatic stages of disease. PMID- 17123872 TI - Co-ordination of DNA single strand break repair. AB - DNA damaging agents generated as a consequence of endogenous metabolism or via exogenous factors can produce a wide variety of lesions in DNA. These include base damage, sites of base loss (abasic sites) and single strand breaks (SSBs). Moreover, reactive oxygen species (ROS) create more diversity by generating SSBs containing modified 3'-ends, such as those containing phosphate, phosphoglycolate and oxidative base damage. Ionising radiation also generates DNA base lesions in close proximity to SSBs. The majority of these non-bulky lesions in DNA are repaired by proteins involved in the base excision repair (BER) pathway. It is apparent that due to the complexity of these lesions, they may require individual subsets of BER proteins for repair. However, the mechanism unravelling the required enzymes and directing damage-specific repair of SSBs is unclear. In this review we will discuss recent studies that identify new enzymes and activities involved in the repair of SSBs containing modified ends and in particular outline the possible mechanisms involved in the co-ordinated repair of "damaged" SSBs that can not be resealed directly and require preliminary processing. PMID- 17123873 TI - Collaborative roles of gammaH2AX and the Rad51 paralog Xrcc3 in homologous recombinational repair. AB - One of the earliest events in the signal transduction cascade that initiates a DNA damage checkpoint is the phosphorylation on serine 139 of histone H2AX (gammaH2AX) at DNA double-strand breaks (DSBs). However, the role of gammaH2AX in DNA repair is poorly understood. To address this question, we generated chicken DT40 cells carrying a serine to alanine mutation at position 139 of H2AX (H2AX( /S139A)) and examined their DNA repair capacity. H2AX(-/S139A) cells exhibited defective homologous recombinational repair (HR) as manifested by delayed Rad51 focus formation following ionizing radiation (IR) and hypersensitivity to the topoisomerase I inhibitor, camptothecin (CPT), which causes DSBs at replication blockage. Deletion of the Rad51 paralog gene, XRCC3, also delays Rad51 focus formation. To test the interaction of Xrcc3 and gammaH2AX, we disrupted XRCC3 in H2AX(-/S139A) cells. XRCC3(-/-)/H2AX(-/S139A) mutants were not viable, although this synthetic lethality was reversed by inserting a transgene that conditionally expresses wild-type H2AX. Upon repression of the wild-type H2AX transgene, XRCC3( /-)/H2AX(-/S139A) cells failed to form Rad51 foci and exhibited markedly increased levels of chromosomal aberrations after CPT treatment. These results indicate that H2AX and XRCC3 act in separate arms of a branched pathway to facilitate Rad51 assembly. PMID- 17123874 TI - Quantification of free and total sialic acid excretion by LC-MS/MS. AB - BACKGROUND: The main purpose for measuring urinary free sialic acid (FSA) is to diagnose sialic acid (SA) storage diseases. Elevated amounts of conjugated sialic acid (CSA) are observed in several diseases indicating the need to quantify CSA as well. A LC-MS/MS method for quantification of FSA and total sialic acid (TSA) in urine is developed and validated. METHODS: FSA is analyzed directly after filtration of urine samples. For determination of TSA an enzymatic (neuraminidase) and a chemical (acid) hydrolysis were compared. 13C3-sialic acid was used as internal standard. LC-MS/MS was performed in negative electrospray ionisation mode with multiple reaction monitoring of transitions m/z 308.2-->87.0 (SA) and m/z 311.2-->90.0 (13C3-SA). CSA was calculated by subtracting FSA from TSA. RESULTS: Limit of detection for FSA and TSA was 0.3 and 1.7 micromol/L, respectively. Limit of quantification for FSA and TSA was 1.0 and 5.0 micromol/L. Intra- and inter-assay variations of FSA were 4.6% and 6.6% (n=10) for FSA and 6.5% and 3.6% (n=10) for TSA. Linearity was tested till 7800 micromol/L (r2=0.9998). Values of SA analyzed after neuraminidase- or acid hydrolysis treatment were comparable. Urine samples from patients with inborn errors of SA (related) metabolism were analyzed and compared with age-related reference values. CONCLUSION: A method has been developed for routine determination of urinary FSA and TSA. The method is rapid, specific, robust and sensitive. Age related reference values for FSA, TSA and CSA were determined and improved diagnostic efficacy. PMID- 17123876 TI - Does QOL provide the same information as toxicity data? PMID- 17123877 TI - Genetics and quality of life. PMID- 17123878 TI - Is the use of QOL data really any different than other medical testing? AB - QOL data and routine clinical data such as laboratory data, whether used in clinical practice or research, can be viewed by clinicians in the same way. Both provide important patient-related information using instruments which are reliable and valid. Both require that the clinician or researcher understand how to utilize and interpret the data and gain comfort with doing so through experience. It is through this repeated usage of QOL data that clinicians will, hopefully, come to accept QOL data as a routine and invaluable part of their practice and view it just as they view other clinical data. Whether it is a clinician looking at QOL data or laboratory data for the first time, or a clinical researcher including QOL data in a clinical trial for the first time, training, repetitive use, and experience is necessary to feel comfortable with integrating the data to recommend an appropriate clinical action or to capture the meaningful effect. PMID- 17123879 TI - Choosing the "correct" assessment tool. PMID- 17123880 TI - Analyzing the "correct" endpoint. AB - The choice of QOL endpoints for a study should be based on which score will most likely change if the treatment is favorable. How the QOL change is calculated should be based on the expected amount of missing data, how many time points data will be collected, and whether extreme outliers in the scores impact results. The study should have sufficient power to detect a meaningful difference between arms (typically 10 points on a 0-100 point scale) in the chosen QOL endpoint. At the conclusion of a study, several secondary endpoints can be analyzed which can provide additional information and confirm primary endpoint results. PMID- 17123881 TI - Dealing with multiple endpoints. PMID- 17123882 TI - Combining symptom and survival data. PMID- 17123883 TI - Integrating QOL assessments for clinical and research purposes. AB - QOL questionnaires can be a useful tool for patients, clinicians, and researchers. Although each has a different goal and investment in the data, each of these perspectives can contribute to truly collaborative care in which the needs of the patient are met. PMID- 17123889 TI - Loss of p27 expression and microsatellite instability in sporadic colorectal cancer. AB - BACKGROUND: The role of the loss of p27 protein expression in the oncogenesis of colorectal cancer is still in debate. In this study, we prospectively examined the immunohistochemical expression of p27 in 108 consecutive colorectal cancers, and we analysed the relationship with the results, the clinicopathological data, microsatellite instability (MSI) and other genetic alterations of tumours. METHODS: Unselected patients (108) who underwent curative colorectal resection for sporadic colorectal cancer in a three-year period were evaluated for MSI using 6 microsatellite markers, and for the presence of p27, p53, Fhit, Mlh1 and Msh2 proteins by means of immunostaining. The relationships between these markers were analysed. p27 protein expression was examined for association with disease recurrences and survival. RESULTS: Lack of p27 expression was noted in 33 out of 108 (30.5%) colorectal cancer cases (P<0.05). This altered expression was significantly higher in proximal cancers (P<0.05), mucinous tumours (P<0.001), poorly differentiated histology (P<0.01), cancers with MSI (P<0.05), tumours with altered expression of Mlh1 (P<0.01), of Msh2 (P<0.05), and of Fhit (P<0.01). Overall survival was better in the patient group with altered level of phenotypic p27 expression, although the difference does not reach statistical significance (P=0.069). The analysis performed only for patients with tumour at stage II showed significantly better survival when the tumour exhibited altered p27 expression (P<0.02). CONCLUSIONS: The results of the present study support the hypothesis that altered expression of p27 may be part of the genetic pathway involving MSI, which is responsible for the development of some colorectal cancers. PMID- 17123890 TI - Shifting goalposts in antibiotic approval. PMID- 17123891 TI - HIV infection as a weapon of war. PMID- 17123892 TI - Nosocomial transmission of community-associated methicillin-resistant Staphylococcus aureus: an emerging threat. PMID- 17123893 TI - Displaced populations and pandemic influenza. PMID- 17123894 TI - Averting avian influenza pandemic: SOS from a developing country. PMID- 17123895 TI - Surveillance for avian influenza in human beings in Thailand. PMID- 17123896 TI - The real alternative therapy for rabies. PMID- 17123897 TI - Scabies: a ubiquitous neglected skin disease. AB - Scabies has been a scourge among human beings for thousands of years. Its worldwide occurrence with epidemics during war, famine, and overcrowding is responsible for an estimated 300 million people currently infested. Scabies refers to the various skin lesions produced by female mites, and their eggs and scybala that are deposited in the epidermis, leading to delayed-type hypersensitivity reaction. Recent immunological findings such as cross-reactivity with house dust mite allergens and an altered T-helper-1/T-helper-2 pattern contribute to a better understanding of the pathomechanism. Furthermore, progress in molecular biology and cloning of relevant antigens could enable the development of a diagnostic ELISA system and candidate vaccines in the near future. Typical and atypical clinical presentations with pruritus as a hallmark of scabies occur in young, pregnant, immunocompromised, and elderly patients and include bullous and crusted (Norwegian) manifestations as well as those masked by steroid use (scabies incognito). This article reviews scabies management strategies in developed countries and resource-poor communities as well as typical complications, including the emergence of resistance and drug-related adverse events. Other problems such as post-scabies eczema and reinfestation, and newer treatments such as ivermectin are also discussed. PMID- 17123898 TI - Malaria in adolescence: burden of disease, consequences, and opportunities for intervention. AB - The problem of malaria in adolescence has largely been overshadowed by the huge burden of disease in young children. A substantial number of adolescents are at risk from malaria infection, but the burden of disease and consequences of infection in this age-group have rarely been studied. Our understanding of specific risk factors and beneficial interventions for adolescents is also limited. Data show that, from an adolescent viewpoint, malaria is a common cause of clinical illness and a preventable cause of death, even in areas of stable malaria transmission. Younger adolescents might be at a higher risk than older adolescents, because of immunological and hormonal factors. There are limited data about the adverse consequences of malaria in non-pregnant adolescents. However, in pregnant adolescents, the consequences of malaria are of great concern and simple interventions might lead to a substantial benefit. Malaria infection in adolescents is an under-recognised problem, and the prevention, diagnosis, and treatment of malaria should have a high priority within adolescent health programmes. PMID- 17123899 TI - HIV-1 infection of mononuclear phagocytic cells: the case for bacterial innate immune deficiency in AIDS. AB - HIV-1 infection of mononuclear phagocytic cells, comprising monocytes, macrophages, and dendritic cells, has been the subject of extensive research over the past 20 years. The roles of mononuclear phagocytic cells in transmission of HIV-1 infection and as reservoirs of actively replicating virus have received particular attention. Experimental data have also accumulated about the effects of HIV-1 on the physiological function of mononuclear phagocytic cells, particularly their role in innate immunity to bacteria. The effects of HIV-1 on bacterial innate immune responses by mononuclear phagocytic cells are discussed here together with reports of direct interactions between HIV-encoded products and bacterial innate immune signalling pathways. These reports demonstrate mechanisms for HIV-mediated disruption of innate immune responses by mononuclear phagocytic cells that could provide novel therapeutic targets in HIV-infected patients. The clinical urgency is highlighted by greatly increased risk of invasive bacterial disease in this population, even in the era of highly active antiretroviral therapy. HIV-mediated injury to bacterial innate immunity provides an experimental paradigm that could broaden our overall understanding of innate immunity and be used to study responses to pathogens other than bacteria. PMID- 17123900 TI - Rotavirus vaccines: considerations for successful implementation in Europe. AB - A group of European experts in infectious diseases and vaccinology has met on several occasions to assess the rationale for universal vaccination against rotavirus infection of infants in Europe. On the basis of the available data, we concluded that vaccination was the best approach to prevent severe rotavirus gastroenteritis, and that European countries should consider implementing rotavirus vaccination in their routine immunisation programmes. The main barrier to the implementation of rotavirus vaccination in Europe is a general lack of awareness of stakeholders, policymakers, health-care professionals, and parents about rotavirus disease and the advantages of vaccination. Further studies on the cost of the disease and the benefit of vaccination, together with raising awareness are necessary steps to ensure successful implementation of rotavirus vaccination in Europe. PMID- 17123901 TI - A case for passive immunoprophylaxis against tuberculosis. AB - HIV-associated tuberculosis is escalating ominously in Africa and southeast Asia despite existing control measures. Therefore, new approaches to tuberculosis control need to be explored. We discuss the potential use of passive immunoprophylaxis with antibodies in tuberculosis control. Although the predominant type of active host resistance is T-cell mediated, recent results in mouse experimental models suggest that monoclonal antibodies to certain antigens (eg, Acr or lipoarabinomannan) can impart substantial passive protection against tuberculous infection. These results are corroborated by data from other laboratories on passive vaccination against a number of intracellular microbial pathogens. Further work is needed to develop human (or humanised) antibody reagents, to increase their protective efficacy, and to expand our understanding of the mechanisms of antibody action. PMID- 17123902 TI - Sir, I turned much darker! PMID- 17123903 TI - Cerebral schistosomiasis. PMID- 17123904 TI - Diversity of the parachlamydiae in the environment. AB - Chlamydiae are obligate intracellular bacteria, parasites of a variety of eukaryotes ranging from amoebae to humans. Among them, the family Parachlamydiaceae comprises endosymbionts of amoebae, mainly Acanthamoeba, currently investigated as emerging pathogens of humans and other vertebrates. 16S rDNA-based PCR culture-independent studies in environmental samples have demonstrated the presence of Chlamydiales in various types of nonmedical habitats. Here we reviewed the biology of the Parachlamydiaceae, and more particularly those studies reporting molecular evidences for their presence in the environment, with a re-analysis of the 16S rDNA phylotypes. PMID- 17123905 TI - High-pressure homogenization as a non-thermal technique for the inactivation of microorganisms. AB - In the pharmaceutical, cosmetic, chemical, and food industries high-pressure homogenization is used for the preparation or stabilization of emulsions and suspensions, or for creating physical changes, such as viscosity changes, in products. Another well-known application is cell disruption of yeasts or bacteria in order to release intracellular products such as recombinant proteins. The development over the last few years of homogenizing equipment that operates at increasingly higher pressures has also stimulated research into the possible application of high-pressure homogenization as a unit process for microbial load reduction of liquid products. Several studies have indicated that gram-negative bacteria are more sensitive to high-pressure homogenization than gram-positive bacteria supporting the widely held belief that high-pressure homogenization kills vegetative bacteria mainly through mechanical disruption. However, controversy exists in the literature regarding the exact cause(s) of cell disruption by high-pressure homogenization. The causes that have been proposed include spatial pressure and velocity gradients, turbulence, cavitation, impact with solid surfaces, and extensional stress. The purpose of this review is to give an overview of the existing literature about microbial inactivation by high pressure homogenization. Particular attention will be devoted to the different proposed microbial inactivation mechanisms. Further, the different parameters that influence the microbial inactivation by high-pressure homogenization will be scrutinized. PMID- 17123906 TI - Candida species adhesion to oral epithelium: factors involved and experimental methodology used. AB - Due to the increasing prevalence and emergence of Non-Candida albicans Candida (NCAC) species, especially in immunosupressed patients, it is becoming urgent to deepen the current knowledge about virulence factors of these species. Adhesion of cells to epithelium is considered one of the major virulence factors of Candida species. However, relatively little is known concerning the adhesion mechanisms of NCAC species to epithelium, as well as about the factors affecting the adhesion process. This review focuses both the mechanisms that regulate the adhesion interactions and the factors involved and the description of the experimental methodology that has been used to perform the adhesion assays. PMID- 17123907 TI - The contribution of cytolethal distending toxin to bacterial pathogenesis. AB - Cytolethal distending toxin (CDT) is a bacterial toxin that initiates a eukaryotic cell cycle block at the G2 stage prior to mitosis. CDT is produced by a number of bacterial pathogens including: Campylobacter species, Escherichia coli, Salmonella enterica serovar Typhi, Shigella dystenteriae, enterohepatic Helicobacter species, Actinobacillus actinomycetemcomitans (the cause of aggressive periodontitis), and Haemophilus ducreyi (the cause of chancroid). The functional toxin is composed of three proteins; CdtB potentiates a cascade leading to cell cycle block, and CdtA and CdtC function as dimeric subunits, which bind CdtB and delivers it to the mammalian cell interior. Once inside the cell, CdtB enters the nucleus and exhibits a DNase I-like activity that results in DNA double-strand breaks. The eukaryotic cell responds to the DNA double strand breaks by initiating a regulatory cascade that results in cell cycle arrest, cellular distension, and cell death. Mutations in CdtABC that cause any of the three subunits to lose function prevent the bacterial cell from inducing cytotoxicity. The result of CDT activity can differ somewhat depending on the eukaryotic cell types affected. Epithelial cells, endothelial cells, and keratinocytes undergo G2 cell cycle arrest, cellular distension, and death; fibroblasts undergo G1 and G2 arrest, cellular distension, and death; and immune cells undergo G2 arrest followed by apoptosis. CDT contributes to pathogenesis by inhibiting both cellular and humoral immunity via apoptosis of immune response cells, and by generating necrosis of epithelial-type cells and fibroblasts involved in the repair of lesions produced by pathogens resulting in slow healing and production of disease symptoms. Thus, CDT may function as a virulence factor in pathogens that produce the toxin. PMID- 17123908 TI - Comparison of platelet pellet and bioactive glass in periodontal regenerative therapy. AB - OBJECTIVE: In recent years, platelet-rich plasma combined with graft materials has been used for periodontal regeneration. The individual role of blood products with guided tissue regeneration in periodontal regenerative therapy is unclear and needs to be elucidated. The purpose of this study was to compare the clinical and radiological effectiveness of platelet pellet/guided tissue regeneration (PP/GTR) and bioactive glass/GTR (BG/GTR) treatments in patients with periodontal disease. MATERIAL AND METHODS: Using a split mouth design, 15 chronic periodontitis patients with pocket depths > or = 6 mm following periodontal initial therapy were randomly assigned to treatment with a combination of PP/GTR or BG/GTR in contralateral dentition areas. An absorbable membrane of polylactic acid was used GTR. The criteria for the comparative study were preoperative and postoperative 6 months pocket depth, clinical attachment level, and radiological alveolar bone level. RESULTS: Both treatment modalities resulted in significant pocket depth reduction and gain in clinical attachment and alveolar bone level compared to the preoperative values (p < 0.01). Reduction in pocket depth, gain in clinical attachment and alveolar bone level were 4(3-6), 4.1+/-0.7, 4.9+/-1.4 mm in the PP/GTR group and 4(3-7), 4.1+/-1.2, 5.9+/-1.7 mm in the BG/GTR group, respectively. The differences between the two groups were not statistically significant (p > 0.05). CONCLUSIONS: Within the limits of this study, it was concluded that PP may be effective as a bioactive glass graft material and used as a graft material for treating intrabony defects. PP thus appears to be a suitable alternative in the regenerative treatment of intrabony periodontal defects. PMID- 17123909 TI - Flowable resin composite as a class II restorative in primary molars: A two-year clinical evaluation. AB - OBJECTIVE: To evaluate the clinical durability of flowable resin composite and resin-modified glass ionomer cement when used as class II restoratives in primary molars. MATERIAL AND METHODS: A total of 190 restorations were placed in 61 children, age in the range of 5-11 years. The restoratives, Tetric Flow, in combination with the adhesives, Excite or Prompt-L-Pop and Vitremer, were used in class II cavities in primary molars. An intra-individual study design was used and the restorations were evaluated by modified USPHS criteria over a 2-year period. RESULTS: 146 of the restorations could be evaluated at 2 years. The cumulative failure rate was 10.6% for Vitremer and 13.6% for Tetric Flow. No statistically significant differences were found in failure rates between different materials or between bonding systems. The main cause of failure for Tetric Flow was secondary caries and for Vitremer wear and dissolution. CONCLUSIONS: Vitremer and Tetric Flow showed no significant difference concerning clinical durability at 2 years when used as class II restoratives in primary molars. Both materials demonstrated acceptable clinical results. PMID- 17123910 TI - Effect of therapeutic jaw exercise on temporomandibular disorders in individuals with chronic whiplash-associated disorders. AB - OBJECTIVE: The aim of this study was to investigate the effect of a specific therapeutic jaw exercise on the temporomandibular disorders of patients with chronic whiplash-associated disorders. MATERIAL AND METHODS: Ninety-four consecutive patients with whiplash-related conditions were referred to and accepted for a treatment period at a center for functional evaluation and rehabilitation during 2001-2002. The patients followed a program of physical therapy, occupational therapy, and pain management. At the start of their stay, they were examined by a physician specialized in rehabilitation medicine and also by a dentist who performed a functional examination of the stomatognathic system. Of the 93 patients who accepted participation in the study, 55 were diagnosed with temporomandibular disorders and chronic whiplash-associated disorders in accordance with the inclusion criteria. They were randomized into a jaw exercise group (n = 25), who performed specific therapeutic jaw exercises, and a control group (n = 30). Both groups undertook the whiplash rehabilitation program at the center. RESULTS: There were no inter- or intra-group differences in symptoms and signs of temporomandibular disorders at baseline, nor at the 3-week and 6-month follow-ups, except for an increase of maximum active mouth-opening capacity in the control group. CONCLUSIONS: In conclusion, the therapeutic jaw exercises, in addition to the regular whiplash rehabilitation program, did not reduce symptoms and signs of temporomandibular disorders in patients with chronic whiplash associated disorders. PMID- 17123911 TI - Clinical aspects and management of bisphosphonates-associated osteonecrosis of the jaws. AB - OBJECTIVE: An increasing incidence of osteonecrosis of the jaws (ONJ) in patients treated with intravenous bisphosphonates has been reported in the literature. The aim of this study was to evaluate the clinical aspects, diagnostic investigations, and management of ONJ associated with bisphosphonates in a series of 12 patients. METHOD: Our patients included 1 asymptomatic and 11 symptomatic subjects. For the symptomatic patients, the osteonecrosis was diagnosed through histological investigations of exposed bone that showed avascular and necrotic tissue with inflammatory infiltrate. The patients were complaining of swelling, fever, and bone exposure involving the jaws. The asymptomatic patient presented as an occasional finding during a routine dental examination and the necrosis was confirmed on the basis of imaging investigations. Radiographic, scintigraphic, and microbiological examinations were carried out for all patients. Treatment included antibiotics, minor surgical interventions, and hyperbaric oxygen therapy. RESULTS: The radiological investigations revealed osteolytic areas and the scintigraphy demonstrated increased bone metabolism. The microbiological analysis showed pathogenic micro-organisms in the majority of patients. Therapy was useful in obtaining short-term symptomatic relief. CONCLUSIONS: Histological, radiological, nuclear medicine, and microbiological investigations are important diagnostic tools for patients with bisphosphonates-associated osteonecrosis of the jaws. However, a long-term follow-up is necessary if we are to better understand the treatment outcome. PMID- 17123912 TI - Antibiotic prescribing practices among Norwegian dentists. AB - OBJECTIVE: There is little information on antibiotic prescribing habits among dentists in general. In 1992 we reported a study among Norwegian dentists, and the present investigation was undertaken to find out if the patterns of antibiotic prescription had changed since then. MATERIAL AND METHODS: A total of 470 randomly selected dentists (10% of total) received a questionnaire and a letter describing the survey and 313 responded. RESULTS: Results indicated that 35% did not issue any prescriptions in a typical week, while 3% issued > or =5. Fifty percent reported that they might prescribe antibiotics when treating periodontal diseases, but only 3.4% reported the use of microbial diagnosis before selecting an antibiotic; 71% of the respondents reported use of antibiotics occasionally to prevent general complications of dental treatment; 80% prescribed antibiotics for prophylactic use if the patient revealed a history of endocarditis, while 5% reported never doing so. CONCLUSION: These findings are in concert with the results obtained 11 years ago, but indicating that dentists who had attended postgraduate courses on antibiotics prescribed such drugs more frequently. This was not statistically significant. However, it is of great concern that 5% never prescribed antibiotics when treating patients with a history of endocarditis, and that 20% did not know that amoxicillin was a penicillin. Such lack of knowledge may cause fatal results of therapy. PMID- 17123913 TI - Effects of isometric contraction on intramuscular level of neuropeptide Y and local pain perception. AB - OBJECTIVE: The release of neuropeptide Y (NPY) is reported to increase in ischemic conditions and may thus be involved in chronic myalgia. The purpose of this study was to investigate the effect of isometric contraction on intramuscular levels of NPY in relation to local pain development. MATERIAL AND METHODS: Intramuscular microdialysis was performed in the masseter and trapezius muscles to determine NPY levels before, during, and after isometric contraction in 16 healthy females. Pain intensity was assessed simultaneously with VAS. Repeated measures ANOVA, t-test, and Pearson correlation analysis were used for statistical analyses. RESULTS: The level of NPY in the trapezius muscle was increased during and after contraction, while there was no change in the masseter muscle. The level of NPY before contraction was higher in the masseter muscle than in the trapezius muscle, and the levels in the two muscles were correlated before and during contraction. Low-level pain in both muscles after probe insertion increased significantly during contraction, but the pain was not correlated to the NPY level. CONCLUSIONS: Pain is developed in the trapezius and masseter muscles during repeated isometric contraction. The NPY level is increased in the trapezius muscle but is not associated with the pain development. PMID- 17123914 TI - Ecological time-trend analysis of caries experience at 12 years of age and caries incidence from age 12 to 18 years: Norway 1985-2004. AB - OBJECTIVES: The purpose of the present investigation was to report on caries experience among Norwegian 12-year-olds from 1985 to 2004 and to assess caries incidence from 12 to 18 years of age for birth cohorts 1973 to 1986. MATERIAL AND METHODS: Aggregated data from the Norwegian Public Dental Services and from official statistics were employed. Information was available about the number of subjects, the proportion receiving treatment, sales of fluoride tablets, socio demographics, caries prevalence, and the number of decayed, missing, and filled teeth (DMFT). RESULTS: An almost linear decline in caries prevalence and mean D3MFT (dentine level) occurred among 12-year-old children from 1985 until the year 2000, but from 2000 to 2004 an increasing trend was observed. The highest mean 6-year D3MFT increment (age 12-18 years) was 4.1 (cohort 1976), while the lowest was 3.2 (cohorts 1982 and 1983). In multiple linear regression analyses of trend, baseline D3MFT accounted for more than 91% of total explained variance in D3MFT increment (Models I and III). Without baseline D3MFT as predictor (Models II and IV), there was a significant association between education, social assistance, mobility, infant mortality, percentage examined, and the additive interaction terms year + income and year + education and D3MFT increment after controlling for confounding and multicollinearity. CONCLUSIONS: Four consecutive years of increase in caries experience among 12-year-old children after 15 years of decline and evidence of stability or increase of the caries increment from 12 to 18 years of age among Norwegian teenagers give cause for concern. PMID- 17123915 TI - Inhibition of mineral loss at the enamel/sealant interface of fissures sealed with fluoride- and non-fluoride containing dental materials in vitro. AB - OBJECTIVE: In this in vitro study we evaluated the enamel mineral loss effect of fluoride-containing and non-fluoride-containing materials at different distances from the sealant margin, and verified the fluoride-releasing capability of these materials. MATERIAL AND METHODS: Extracted molars were randomly assigned into nine groups (n = 12): Concise (C), FluroShield (F), Helioseal Clear Chroma (H), Vitremer (V), Fuji II-LC (FII), Ketac Molar (KM), Fuji IX (FIX), Single Bond (SB), and Clearfil Protect Bond (CF). All groups were subjected to thermo and pH cycling. Enamel mineral loss was evaluated by cross-section micro-hardness analysis at distances: -100 microm, 0 microm, 100 microm, 200 microm. The mineral loss data were analyzed using a multi-factor ANOVA with split-plot design, and fluoride-released data were submitted to ANOVA and Tukey tests. RESULTS: FIX demonstrated a lower mineral loss than C, F, and H, but did not differ from the SB, CF, V, FII, and KM groups, which also demonstrated no difference among them. C, F, H, and V presented the highest mineral loss, with no difference among them. V did not differ from the other groups (p > 0.05). Regarding the different distances from the sealant margin, -100 microm presented the lowest mineral loss. FIX showed the highest fluoride release on the 7th and 14th days of evaluation, while CF showed high fluoride release only on the 7th day. CONCLUSION: Resin sealant did not prevent enamel mineral loss, contrary to glass-ionomer cement, which showed the highest capacity for fluoride release. It is not exclusively the presence of fluoride in a material's composition that indicates its capability to interfere with the development of enamel caries-like lesions. PMID- 17123916 TI - Effect of continuous longitudinal glass fiber reinforcement on the cantilever beam strength of particulate filler composites. AB - OBJECTIVE: The objective of this in vitro study was to investigate the effect of continuous longitudinal glass fibers on the bending properties of particulate filler composite resins. MATERIAL AND METHODS: Four particulate filler composite resins (Tetric Ceram, Point 4, Z250, P60) and one continuous longitudinal glass fiber material (everStick) as reinforcement were selected. Three groups of beam specimens (5 x 5 x 25 mm) for each material were fabricated. For group I, specimens were not reinforced with fibers and were tested as the control group. For group II, specimens were reinforced with 2 layers of the fiber (6.5 vol%), and for group III with 3 layers (9.8 vol%). The specimens were stored at 37 degrees C for 30 days. One end of the beam specimen (14 mm) was fixed in a device, while the other (11 mm) was left free. Cantilever beam strength was measured using a universal testing machine, loading at a distance of 10 mm from the upright support through a steel ball of 2 mm diameter. The loads and deflection at initial failure and at final failure for each specimen were recorded. The data were statistically analyzed using one-way ANOVA and the multiple comparison Scheffe test (alpha = 0.05). RESULTS: The bending moments of particulate filler composite at final failure, when reinforced with 3 layers of the glass fibers (272.4-325.2 Ncm), were significantly higher than for the composites without fibers. However, the materials reinforced with 3 layers of fibers were not significantly different from the materials reinforced with 2 layers of fibers (234.1-282.6 Ncm). The materials reinforced with 3 layers of fibers exhibited severe deflection at final failure, ranging from 2.8 mm to 3.4 mm. The bending moments of the particulate composites increased linearly with the weight fraction of the fillers, but there was no linear correlation between them when reinforced with fibers. CONCLUSIONS: The cantilever beam strength of the particulate filler composites increased significantly when layers of fibers were added and as the weight fraction of filler increased, but a higher fiber volume fraction did not lead to a significantly higher cantilever beam strength. PMID- 17123917 TI - Wax encapsulation of water-soluble compounds for application in foods. AB - Water-soluble ingredients have been successfully encapsulated in wax using two preparation techniques. The first technique ('solid preparation') leads to relatively large wax particles. The second technique ('liquid preparation') leads to relatively small wax particles immersed in vegetable oil. On the first technique: stable encapsulation of water-soluble colourants (dissolved at low concentration in water) has been achieved making use of beeswax and PGPR. The leakage from the capsules, for instance of size 2 mm, is about 30% after 16 weeks storage in water at room temperature. To form such capsules a minimum wax mass of 40% relative to the total mass is needed. High amounts of salt or acids at the inside water phase causes more leaking, probably because of the osmotic pressure difference. Osmotic matching of inner and outer phase can lead to a dramatic reduction in leakage. Fat capsules are less suitable to incorporate water soluble colourants. The reason for this could be a difference in crystal structure (fat is less ductile and more brittle). On the second technique: stable encapsulation of water-soluble colourants (encapsulated in solid wax particles) has been achieved making use of carnauba wax. The leakage from the capsules, for instance of size 250 mm, is about 40% after 1 weeks storage in water at room temperature. PMID- 17123918 TI - Controlled delivery of a hydrophilic drug from a biodegradable microsphere system by supercritical anti-solvent precipitation technique. AB - The purpose of this study was to prepare microspheres loaded with hydrophilic drug, bupivacaine HCl using poly(D,L-lactic-co-glycolic acid) (PLGA) and poly(L lactic acid) (PLLA). Microspheres were prepared with varying the PLGA/PLLA ratio with two different levels of bupivacaine HCl (5 and 10%) using a supercritical anti-solvent (SAS) technique. Microspheres ranging from 4-10 microm in geometric mean diameter could be prepared, with high loading efficiency. Powder X-ray diffraction (PXRD) revealed that bupivacaine HCl retained its crystalline state within the polymer and was present as a dispersion within the polymer phase after SAS processing. The release of bupivacaine HCl from biodegradable polymer microspheres was rapid up to 4 h, thereafter bupivacaine HCl was continuously and slowly released for at least 7 days according to the PLGA/PLLA ratio and the molecular weight of PLLA. PMID- 17123919 TI - Enhancing drug incorporation into tetracycline-loaded chitosan microspheres for periodontal therapy. AB - PURPOSE: To identify optimal formulation parameters for enhancing the incorporation of tetracycline hydrochloride into chitosan microspheres for periodontal therapy. METHODS: Tetracycline-loaded chitosan microspheres were prepared by ionotropic gelation. Various formulation parameters (salt form of drug, aqueous phase pH, anion structure, inorganic salts and electrolytes, preparation method) were investigated for their influence on drug incorporation efficiency. Microspheres were assessed in terms of drug entrapment and content, microsphere recovery, particle size and morphology. RESULTS: Although drug incorporation efficiency was increased marginally, the use of a dihydrate form of the drug was not considered feasible due to the lowered microsphere recovery and higher costs. A decrease in the aqueous pH from 9 to 6 enhanced drug incorporation efficiency without an adverse effect on microsphere morphology. The use of inorganic salts/electrolytes and other approaches of microsphere preparation did not significantly enhance drug incorporation efficiency and these approaches also adversely affected microsphere morphology. The ionotropic preparation method in terms of the drug loading technique significantly affected drug incorporation efficiencies. CONCLUSIONS: This study has shown that formulation variables can be exploited in order to enhance the incorporation of a water soluble drug into chitosan microspheres using the ionotropic gelation technique. Based on a comparison of all results obtained with the different approaches, the modification of the aqueous phase to pH 6 was identified as the most feasible approach. PMID- 17123920 TI - Electrophoretic ink using urea-formaldehyde microspheres. AB - A kind of electronic ink, which is supported on indium-tin-oxide (ITO) glass before polymerization, is prepared using urea and formaldehyde resin as wall materials, in which TiO(2) modified with PMMA are dispersed in tetrachloroethylene (TCE) using a mixture of oil blue dyes and charge control additive (span80). Rotary viscometer, Fourier transform infrared spectroscopy (FT IR) and optical microscopy are used to characterize the particles, respectively. The electrophoretic mobility of the microcapsules to electric field is also investigated with two parallel electrodes. PMID- 17123921 TI - Antibody-conjugated soybean oil-filled calcium phosphate nanoshells for targetted delivery of hydrophobic molecules. AB - Hollow calcium phosphate nanoparticles capable of encapsulating poorly water soluble molecules were produced by self-assembly. Previously reported were solid calcium phosphate nanoparticles and water-filled calcium phosphate nanocapsules suited for encapsulating mostly hydrophilic, but not hydrophobic compounds. Here, calcium phosphate was deposited around 100 nm diameter, 1,2-dioleoyl-sn-glycero-3 phosphate stabilized soybean oil nanoemulsions using either calcium chloride or NaOH titrations to achieve shell thickness between 20-70 nm. The surface was functionalized with carboxylic acid via the addition of carboxyethylphosphonic acid to attach Molecular Probes AB-594C antibody using sulpho-n hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride with an efficiency of approximately 70%, while retaining near complete antibody function. Hydrophobic pyrene was encapsulated with an efficiency of 95%, at concentrations much higher than its water solubility limit, and exhibited spectral features characteristic of a hydrophobic environment. These materials can be used in the targeted delivery of many useful, yet poorly water-soluble pharmaceutical and nutraceutical compounds. PMID- 17123922 TI - Biodegradable micro- and nanoparticles as long-term delivery vehicles for gentamicin. AB - Micro- and nanoparticles of poly(lactide-co-glycolide) (PLGA) loading gentamicin were prepared by a solvent evaporation method with the aim of obtaining appropriate vectors for systemic administration. Microspheres presented mean diameters below 3 microm and nanoparticles showed homogeneous sizes with a diameter of 320 nm. Drug loading was more efficient in the case of microencapsulation. The more hydrophilic copolymers with carboxyl-end groups yielded higher microparticle loadings, reaching encapsulation efficiencies up to 9.2 microg mg(-1) of polymer (502H, 503H or 75:25H). Nanoparticles made of 502H PLGA also achieved an acceptable level of encapsulation (6.2 microg mg(-1)). Particles prepared by using the solvent evaporation method showed no aggregation after hydration, in contrast to the microparticles prepared by spray-drying which showed fast and high auto-aggregation. In vitro release profiles revealed that 503H microspheres showed the highest burst during the first hour, while the most sustained release was for microparticles of 502H copolymer (40% of gentamicin remained in the formulation after 28 days). In summary, microspheres made of 502H, 503H and 75:25H and nanoparticles of 502H showed the best potential properties for systemic use in the treatment of intra-cellular gentamicin susceptible pathogens. PMID- 17123923 TI - Rotary spray congealing of a suspension: effect of disk speed and dispersed particle properties. AB - Microspheres were produced by rotary spray congealing molten suspensions composed of poloxamer 188 and a single solid excipient. Six different excipients, spanning a range of particle size, density and morphology were used. The size distribution, true density and shape of the resulting congealed particles were assessed. Disk rotation speed had the most significant effect on microsphere size, with higher speeds yielding smaller congealed particles. At low disk speeds, congealed particle size decreases with increasing dispersed solid density. Over the range investigated, neither suspended solid size nor morphology had a significant effect on the congealed microsphere size. Measurements of the density of the microspheres suggest that the solids were homogeneously spread across the different congealed particle sizes. This suggests that rotary atomization of poloxamer 188 suspensions is an effective method for producing spherical particles for encapsulation for a wide variety of drug particle sizes, shapes and densities--with disk speed being used to control the mean size. PMID- 17123927 TI - 'Participate to learn': a promising practice for community ABI rehabilitation. AB - OBJECTIVE: To identify best practices and promising practices to enhance participation in meaningful and productive activities. METHOD: An electronic search of the ABI rehabilitation research literature since 1990 yielded 974 articles of which 30 focused on interventions that targeted participation and evaluated effectiveness using direct measures of participation. Three reviewers rated these articles according to the standards set out by the Centre for Reviews and Dissemination. Following the systematic review, an interpretive review of the same articles was completed. RESULTS: Only three studies were rated as strong. No best practices were identified. Three promising practices found some support. The interpretive review suggested 'Participate to learn' as a useful rehabilitation model. The model rests on roles as goals, learning by experience in real-life contexts and the use of personal and environmental support to enable participation. CONCLUSIONS: 'Participate to learn' is both a credible rehabilitation model and deserving of more study. PMID- 17123928 TI - Effectiveness of vocational rehabilitation following acquired brain injury: preliminary evaluation of a UK specialist rehabilitation programme. AB - PRIMARY OBJECTIVE: To report the outcome of 232 clients undertaking Rehab UKs Vocational Rehabilitation Programme, over three consecutive years, in three centres across the UK. METHODS AND PROCEDURES: Data regarding demographic characteristics of participants, type and severity of acquired brain injury were collected on admission to the programme. The duration of the vocational programme and occupational status on discharge is also reported. INTERVENTIONS: A description of the vocational programme is given including selection and admission criteria, main components of the programme, monitoring and evaluation frameworks. RESULTS: Forty-one per cent of participants were discharged into paid competitive employment, with a further 16% gaining voluntary work and 15% taking up mainstream training or education. The remaining 28% were referred to other services or withdrew from the programme. CONCLUSIONS: Vocational rehabilitation offering educational and experiential learning opportunities is effective in enabling participants with severe acquired brain injuries to return to paid employment. PMID- 17123929 TI - Patients with mild traumatic brain injury: immediate and long-term outcome compared to intra-cranial injuries on CT scan. AB - BACKGROUND: Mild traumatic brain injury (MTBI) defined as Glasgow Coma Scale (GCS) 14 or 15 has shown contradictory short- and long-term outcomes. The objective of this study was to correlate intra-cranial injuries (ICI) on CT scan to neurocognitive tests at admission and to complaints after 1 year. METHODS: Two hundred and five patients with MTBI underwent a CT scan and were examined with neurocognitive tests. After 1 year complaints were assessed by phone interviews. RESULTS: The neurocognitive tests in 51% of the patients showed significant deficits; there was no difference for patients with GCS 14-15, nor was there a difference between patients with ICI to patients without. After 1 year patients with ICI had significantly more complaints than patients without ICI, the most frequent complaint was headache and memory deficits. CONCLUSIONS: No correlation was found between GCS or ICI and the neurocognitive tests upon admission. After 1 year, patients with ICI have significantly more complaints than patients without ICI. No cost savings resulted by doing immediate CT scan on all. PMID- 17123930 TI - Neuropsychological profile of persons with anoxic brain injury: differences regarding physiopathological mechanism. AB - PRIMARY OBJECTIVE: To determine the neuropsychological profile of persons with anoxic brain injury. METHODS AND PROCEDURES: A retrospective study on a population of persons with anoxic brain injury admitted to a Brain Injury Unit (Institut Guttmann, Spain) from 1995-2003. The sample was divided according to physiopathological mechanisms in two sub-groups: ischemic anoxia (21 cases) and hypoxemic anoxia (11 cases). Functions assessed included orientation, attention, language, visuo-perceptive and visuo-constructive processing and verbal memory. RESULTS: Neuropsychological assessment showed diffuse cognitive impairment in all assessed functions. Episodes of ischemic anoxia caused more severe verbal memory and learning problems than episodes of hypoxemic anoxia. CONCLUSIONS: This study shows that memory problems are the most prominent and relevant impairment, although all other cognitive functions are also impaired, affecting both memory itself and general behaviour. Statistical analysis also provides preliminary evidence on the different profile of memory impairment whether cerebral anoxia had hypoxic or ischemic origin. PMID- 17123931 TI - Determining long-term symptoms following mild traumatic brain injury: method of interview affects self-report. AB - PRIMARY OBJECTIVE: To examine the role played by two interviewing methods used (spontaneous response and suggested response) in the evaluation of long-term subjective post-mild traumatic brain injury (mTBI) symptoms. RESEARCH DESIGN: Cohort study. METHOD AND PROCEDURES: One hundred and eight adult participants were contacted for a follow-up telephone interview 12-36 months after their mTBI. The participants had to firstly spontaneously indicate symptoms that were still present following their mTBI (spontaneous response). Secondly, a list of symptoms was read to the participants and they had to say whether or not they were afflicted by each symptom (suggested response). Paired t-tests were performed to compare the means obtained using the two methods. The percentage of symptoms reported with the two interviewing methods were used to analyse symptom types. EXPERIMENTAL INTERVENTION: None. MAIN OUTCOMES AND RESULTS: Results show that participants reported significantly more symptoms and a given symptom when a list was read to the participants. Furthermore, neither the number of symptoms nor the type of symptoms reported is identical for the two interviewing methods. CONCLUSION: The interviewing method used influences the number and type of long term post-mTBI symptoms reported by participants. PMID- 17123932 TI - Masculine role adherence and outcomes among men with traumatic brain injury. AB - PRIMARY OBJECTIVE: Traumatic brain injury (TBI) is a significant health problem disproportionately affecting men and is often associated with changes in masculine role functioning in life domains such as vocational functioning, sexual and inter-personal functioning and personal independence. These changes could have serious implications for men's adjustment following injury. The aim of this study was to examine the relations among traditional masculine role adherence, psychosocial adjustment and rehabilitation outcomes in men with TBI. RESEARCH DESIGN: A correlational design was chosen to examine the relations among variables. Spearman correlations and Wilcoxon Rank Sum tests were used to examine relationships between masculine role variables and outcome variables. METHODS AND PROCEDURES: The study included 33 men with TBI who had been discharged from inpatient rehabilitation within 5 years. Participants completed surveys on traditional masculine gender role adherence and gender role conflict and additional data, including measures of functional outcome, life satisfaction, psychosocial outcomes and earnings, were obtained through the TBI Model System longitudinal data collection system. MAIN OUTCOMES AND RESULTS: The results revealed significant associations between masculine role adherence and satisfaction with life, follow-up earnings and FIM change from admission to discharge. CONCLUSIONS: In the current study, particular masculine role variables corresponded to different functional and psychological outcomes. Understanding these differences provides new directions for treatment and offers important information about aspects of traditional masculine roles that may enhance or hinder adjustment to injury. PMID- 17123933 TI - Aggression after traumatic brain injury: analysing socially desirable responses and the nature of aggressive traits. AB - PRIMARY OBJECTIVE: To compare patients with traumatic brain injury (TBI) with controls on sub-types of aggression and explore the role of social desirability. DESIGN: Quasi-experimental, matched-participants design. METHODS AND PROCEDURES: Sixty-nine participants were included in the study. The sample comprised a TBI group (n = 24), a spinal cord injury (SCI) group (n = 21) and an uninjured (UI) group of matched healthy volunteers (n = 24). Participants were given self-report measures of aggression, social desirability and impulsivity. Sixty-one independent 'other-raters' were nominated, who rated participant pre-morbid and post-morbid aggression. MAIN OUTCOMES AND RESULTS: Using standardized norms, 25 39% of participants with TBI were classified as high average-very high on anger and 35-38% as high average-very high on verbal aggression. Other-raters rated participants with TBI as significantly higher on verbal aggression than SCI and UI participants. There were no differences between the groups on physical aggression. The TBI group also had higher levels of impulsivity than SCI and UI groups. Social desirability was a highly significant predictor of self-reported aggression for the entire sample. CONCLUSIONS: Impulsive verbal aggression and anger are the principal aggressive traits after brain injury. Physical aggression may present in extreme cases after TBI, but appears less prominent overall in this population. Social desirability, previously overlooked in research examining TBI aggression, emerged as an influential variable that should be considered in future TBI research. PMID- 17123934 TI - Agitation following traumatic brain injury: an Australian sample. AB - PURPOSE: Agitation following traumatic brain injury (TBI) is perceived to limit the achievement of rehabilitation goals. The aims of this study were: (1) To examine the nature and incidence of agitation during TBI rehabilitation in an Australian sample; and (2) To explore the relationship between agitation and functional outcomes after TBI. METHOD: Retrospective medical record review of 80 participants admitted for rehabilitation following TBI. Outcome data collected at discharge, 6 and 24 months post-discharge, included length of stay, duration of post-traumatic amnesia (PTA), community integration, functional independence and life satisfaction. RESULTS: Seventy per cent of patients demonstrated agitation during rehabilitation for an average of 32 days. Agitated patients experienced longer PTA duration, increased length of stay and reduced functional independence, specifically cognitive function, at discharge. These differences between agitated and non-agitated participants did not appear to persist and were not evident at follow-up 6 and 24 months post-discharge. Longer duration of agitation was associated with delayed PTA emergence, longer rehabilitation stay and reduced functional independence at discharge and follow-up. CONCLUSIONS: A large proportion of patients with TBI experience agitation. Presence of agitation seems to limit rehabilitation gains at discharge. Longer duration of agitation is associated with persisting limitations to functional independence. PMID- 17123935 TI - Outpatient rehabilitation of patients with chronic cognitive impairments after ruptured anterior communicating artery aneurysms reduces the burden of care: a pilot study. AB - PRIMARY OBJECTIVE: To describe the functional outcome of 19 patients with anterior communicating artery aneurysm following the completion of an inter disciplinary out-patient rehabilitation programme. RESEARCH DESIGN: Retrospective descriptive study. SETTING: Community-based inter-disciplinary outpatient programme. PATIENTS: n = 19; consecutive referred sample, mean 182 days post onset; in-patient rehabilitation completed. INTERVENTION: Inter-disciplinary treatment of functional activities; 2-5 hours/day, 3-5 days/week; mean duration: 55-57 days. MAIN OUTCOME MEASURES: Supervision rating scale (SRS) and change in prevalence at admission and discharge of executive impairments, memory, confabulation, apathy, initiation, social inappropriateness and incontinence. RESULTS: Sixty per cent of the patients showed a clinically significant improvement in their SRS from requiring full-time supervision to part-time supervision. Change in SRS was correlated with change in the impairments of executive function, memory and confabulation. CONCLUSION: Although pervasive impairments associated with this disorder may limit capacity for even moderate independence, substantial reduction in direct supervision by family members may be achieved. PMID- 17123936 TI - An evaluation of the paediatric acquired brain injury community outreach programme (PABICOP). AB - PRIMARY OBJECTIVE: To examine the utility of a coordinated, family/community focused programme (PABICOP) vs. a standard approach for improving outcomes for children with ABI and their families. RESEARCH DESIGN: Pre-test-post-test design, with comparison group and follow-up. METHODS AND PROCEDURES: Ninety-six children (64 children receiving PABICOP services and 32 children receiving standard care) participated in the study. Measures were completed at baseline and 3 and 12 months later. MAIN OUTCOMES AND RESULTS: Parents/caregivers with more than 10 contacts with PABICOP scored significantly higher on an ABI knowledge quiz than either parents/caregivers with 10 contacts or less or the comparison group at post-test and follow-up. Parents/caregivers with 10 contacts or less with PABICOP reported significantly greater improvements in children's school and total competence on the CBCL than either parents/caregivers with more than 10 contacts or the comparison group at post-test and follow-up. CONCLUSIONS: PABICOP may be more useful for enhancing knowledge of ABI for parents/caregivers and for integrating children into the community over a 1-year period than a standard approach. The amount of service received appears to influence outcomes. PMID- 17123937 TI - Rehabilitation of attention in two patients with traumatic brain injury by means of 'attention process training'. AB - PRIMARY OBJECTIVE: This case study aims to evaluate the effectiveness of the Solhberg and Mateer's Attention Process Training (APT) using a comprehensive evaluation of various attentional processes. METHODS AND PROCEDURES: Two patients with severe traumatic brain injury were given the APT in a chronic phase. Attentional processes were evaluated at various stages before, during and after treatment, using the Testbatterie zur Aufmerksamkeitsprufung and the Test of everyday attention. MAIN OUTCOMES AND RESULTS: Both patients showed some degree of recovery, particularly in attentional tasks with a selective component. Lesser improvement was present in the case of tasks mapping on the intensity dimension of attention (alertness, vigilance). Training achievements were confirmed by the use of a functional scale evaluating attention, pointing to the generality of improvements. CONCLUSION: The results indicate selective training effects of APT on the attentional disturbances of TBI patients. PMID- 17123938 TI - Report on the typhus epidemic in Upper Silesia. 1848. PMID- 17123939 TI - Insulin-like growth factors and their binding proteins define specific phases of myometrial differentiation during pregnancy in the rat. AB - While the insulin-like growth factor (IGF) system is known to regulate uterine function during the estrous cycle, there are limited data on its role in myometrial growth and development during pregnancy. To address this issue, we defined the expression of the Igf hormones (1 and 2), their binding proteins (Igfbp 1-6), and Igf1r receptor genes in pregnant, laboring, and postpartum rat myometrium by real-time PCR. IGF family genes were differentially expressed throughout gestation. Igf1 and Igfbp1 mRNA levels were upregulated during proliferative phase (Days 6-12) of rat gestation. Igfbp3 gene expression also was elevated in proliferating smooth muscle cells (SMCs) and was highest at the time of transition between proliferative and synthetic phases (Days 12-15). Igfbp6 gene expression profile paralleled plasma progesterone (P4) concentrations, peaking during the synthetic phase (Days 17-19) and decreasing thereafter. Administration of P4 at late pregnancy (starting from Day 20) to maintain elevated plasma P4 concentrations blocked the onset of labor and prevented the fall in Igfbp6 mRNA levels. In contrast, the treatment of pregnant rats with the P4 receptor antagonist RU486 on Day 19 induced preterm labor and the premature decrease of Igfbp6 gene expression. Igfbp2 gene expression was transiently upregulated during the contractile phase of gestation (Days 21-23) solely in the gravid horn of unilaterally pregnant rats, but it was not affected in P4- or RU486-treated animals, supporting a role for mechanical stretch imposed by the growing fetuses. Igfbp5 gene was induced during postpartum involution. Our results suggest the importance of the IGF system in phenotypic and functional changes of myometrial SMCs throughout gestation in preparation for labor. PMID- 17123940 TI - Semen-coagulating protein, SVS2, in mouse seminal plasma controls sperm fertility. AB - Mammalian seminal plasma is known to contain a decapacitation factor(s) that prevents capacitation and thus, the fertility of sperm. This phenomenon has been observed in experiments conducted in vitro that assessed the inhibition of epididymal sperm fertility by seminal plasma or by the purified decapacitation factor. However, the phenomenon of decapacitation has not yet been characterized in vivo. In the present study, we demonstrate that seminal vesicle protein secretion 2 (SVS2), which is a 40-kDa basic protein and a major component of the copulatory plug, enters the uterus and interacts with ejaculated sperm heads after copulation. The SVS2-binding region of sperm changed from the postacrosomal region to the equatorial segment, while the sperm migrated through the uterus and finally disappeared in the oviduct. Furthermore, SVS2 reduced the fertility of epididymal sperm. The sperm treated with SVS2 decreased the percentage of fertilized oocytes from 60% to 10%. The capacitation state was assessed by protein tyrosine phosphorylation and the comprehensiveness of the acrosome reaction. SVS2 functioned to maintain sperm in the uncapacitated state and to reverse capacitated sperm to the uncapacitated state. We found that the fertility of ejaculated sperm is associated with SVS2 distribution in the female reproductive tract. These results indicate that SVS2 functions as a decapacitation factor for mouse sperm. PMID- 17123941 TI - Receptor-determined susceptibility of preimplantation embryos to pseudorabies virus and porcine reproductive and respiratory syndrome virus. AB - In the present study, the in vitro interaction of embryos with pseudorabies virus (PRV) and porcine reproductive and respiratory syndrome virus (PRRSV) was investigated by viral antigen detection and by evaluating the expression of virus receptors, namely, poliovirus receptor-related 1 (PVRL1; formerly known as nectin 1) for PRV and sialoadhesin for PRRSV. Embryonic cells of zona pellucida intact embryos incubated with PRV remained negative for viral antigens. Also, no antigen positive cells could be detected after PRV incubation of protease-treated embryos, since the protease disrupted the expression of PRVL1. However, starting from the five-cell-stage onwards, viral antigen-positive cells were detected after subzonal microinjection of PRV. At this stage, the first foci of PVRL1, also a known cell adhesion molecule, were expressed. At the expanded blastocyst stage, a lining pattern of PVRL1 in the apicolateral border of trophectoderm cells was present, whereas the expression in the inner cell mass was low. Furthermore, PVRL1-specific monoclonal antibody CK41 significantly blocked PRV infection of trophectoderm cells of hatched blastocysts, while the infection of the inner cell mass was only partly inhibited. Viral antigen-positive cells were never detected after PRRSV exposure of preimplantation embryos up to the hatched blastocyst stage. Also, expression of sialoadhesin in these embryonic stages was not detected. We conclude that the use of protease to investigate the virus embryo interaction can lead to misinterpretation of results. Results also show that blastomeres of five-cell embryos up to the hatched blastocysts can become infected with PRV, but there is no risk of a PRRSV infection. PMID- 17123942 TI - Effects of metformin on bovine granulosa cells steroidogenesis: possible involvement of adenosine 5' monophosphate-activated protein kinase (AMPK). AB - In mammals, IGFs are important for the proliferation and steroidogenesis of ovarian cells. Metformin is an insulin sensitizer molecule used for the treatment of the infertility of women with polycystic ovary syndrome. It is, however, unclear whether metformin acts on ovarian cells. Adenosine 5' monophosphate activated protein kinase (AMPK) is involved in metformin action in various cell types. We investigated the effects of metformin on bovine granulosa cell steroidogenesis in response to IGF1 and FSH, and studied AMPK in bovine ovaries. In granulosa cells from small follicles, metformin (10 mM) reduced production of both progesterone and estradiol and decreased the abundance of HSD3B, CYP11A1, and STAR proteins in presence or absence of FSH (10(-8) M) and IGF1 (10(-8) M). In cows, the different subunits of AMPK are expressed in various ovarian cells including granulosa and theca cells, corpus luteum, and oocytes. In bovine granulosa cells from small follicles, metformin, like AICAR (1 mM) a pharmaceutical activator of AMPK, increased phosphorylation of both Thr172 of AMPK alpha and Ser 79 of ACACA (Acetyl-CoA Carboxylase). Both metformin and AICAR treatment reduced progesterone and estradiol secretion in presence or absence of FSH and IGF1. Metformin decreased phosphorylation levels of MAPK3/MAPK1 and MAPK14 in a dose- and time-dependent manner. The adenovirus-mediated production of dominant negative AMPK abolished the effects of metformin on secretion of progesterone and estradiol and on MAPK3/MAPK1 phosphorylation but not on MAPK14 phosphorylation. Thus, in bovine granulosa cells, metformin decreases steroidogenesis and MAPK3/MAPK1 phosphorylation through AMPK activation. PMID- 17123943 TI - Induction of epididymal boar sperm capacitation by pB1 and BSP-A1/-A2 proteins, members of the BSP protein family. AB - A family of proteins designated BSP-A1, BSP-A2, BSP-A3, and BSP-30-kDa, collectively called BSP (bovine seminal plasma) proteins, constitute the major protein fraction of bull seminal plasma. BSP proteins can stimulate sperm capacitation by inducing cholesterol and phospholipid efflux from sperm. Boar seminal plasma contains one homologous protein of the BSP family, named pB1; however, its physiological role is still unknown. In the current study, we report a novel method to purify pB1 from boar seminal plasma by chondroitin sulfate B affinity chromatography and reverse-phase-high performance liquid chromatography. We also studied the effect of pB1, BSP-A1/-A2, and whole boar seminal plasma on boar sperm capacitation. Boar epididymal sperm were washed, preincubated in noncapacitating medium containing pB1 (0, 2.5, 5, 10 or 20 microg/ml), BSP-A1/-A2 (0 or 20 microg/ml) proteins, or whole seminal plasma (0, 250, 500, or 1000 microg/ml), then washed and incubated in capacitating medium. Acrosomal integrity was assessed by chlortetracycline staining. The status of sperm capacitation was evaluated by the capacity of sperm to undergo the acrosome reaction initiated by the addition of the calcium ionophore, A23187. The pB1 and BSP-A1/-A2 proteins increased epididymal sperm capacitation as compared with control (sperm preincubated without proteins). This effect reached a maximum level at 10 microg/ml pB1 and at 20 microg/ml BSP-A1/-A2 (2.3- and 2.2-fold higher than control, respectively). Whole boar seminal plasma did not induce sperm capacitation. In addition, pB1 bound to boar epididymal sperm and was lost during capacitation. These results indicate that BSP proteins and their homologs in other species induce sperm capacitation in a similar way. PMID- 17123944 TI - The 193-base pair Gsg2 (haspin) promoter region regulates germ cell-specific expression bidirectionally and synchronously. AB - Haspin is a unique protein kinase expressed predominantly in haploid male germ cells. The genomic structure of haspin (Gsg2) has revealed it to be intronless, and the entire transcription unit is in an intron of the integrin alphaE (Itgae) gene. Transcription occurs from a bidirectional promoter that also generates an alternatively spliced integrin alphaE-derived mRNA (Aed). In mice, the testis specific alternative splicing of Aed is expressed bidirectionally downstream from the Gsg2 transcription initiation site, and a segment consisting of 26 bp transcribes both genomic DNA strands between Gsg2 and the Aed transcription initiation sites. To investigate the mechanisms for this unique gene regulation, we cloned and characterized the Gsg2 promoter region. The 193-bp genomic fragment from the 5' end of the Gsg2 and Aed genes, fused with EGFP and DsRed genes, drove the expression of both proteins in haploid germ cells of transgenic mice. This promoter element contained only a GC-rich sequence, and not the previously reported DNA sequences known to bind various transcription factors--with the exception of E2F1, TCFAP2A1 (AP2), and SP1. Here, we show that the 193-bp DNA sequence is sufficient for the specific, bidirectional, and synchronous expression in germ cells in the testis. We also demonstrate the existence of germ cell nuclear factors specifically bound to the promoter sequence. This activity may be regulated by binding to the promoter sequence with germ cell-specific nuclear complex(es) without regulation via DNA methylation. PMID- 17123945 TI - NF-kappaB activation elicited by ionizing radiation is proapoptotic in testis. AB - The transcription factor NF-kappaB modulates apoptotic machinery following activation by the IkappaB kinase (IKK) complex. Inhibiting activity of one of the catalytic subunits of the IKK complex, IKKbeta (also known as IKBKB and IKK2) severely inhibits NF-kappaB nuclear translocation in response to most stimuli, including ionizing radiation. Doubly floxed Ikkbeta(F/F) mice (control) were compared to haplo-insufficient Ikkbeta(F/)(delta) mice (NF-kappaB knockdown) to examine the in vivo apoptotic role of NF-kappaB in the testis. Although Ikkbeta(F/F) control adult mice had spermatid head counts and testis and body weights similar to Ikkbeta(F/)(delta) mice, cellular stress in the form of ionizing radiation elicited a differential phenotype. Lower body exposure to 5 Gy of ionizing radiation induced a greater NF-kappaB activation in Ikkbeta(F/F) than in Ikkbeta(F/)(delta) mice. In addition, exposure to ionizing radiation resulted in fewer apoptotic germ cells 3, 6, and 12 h after injury in NF-kappaB knockdown mice than in controls, concomitant with the reduced cleavage of caspases 3 and 9 at 3 h. There was also a reduction in total germ cells lost after radiation with NF-kappaB inhibition. Correspondingly, real-time RT-PCR showed a significant reduction in Cdnk1a (also known as p21) and Fasl expression 3 and 6 h, respectively, after irradiation in Ikkbeta(F/)(delta) compared to control testes. These data indicate that, despite acting in an antiapoptotic manner in many tissue types, NF-kappaB is proapoptotic in modulating the germ cell response to ionizing radiation. PMID- 17123946 TI - Reproductive deficits in male freshwater turtle Chrysemys picta from Cape Cod, Massachusetts. AB - Contaminated groundwater plumes have formed on the Massachusetts Military Reservation (MMR), a Superfund site on Cape Cod, Massachusetts, as a result of chemical waste disposal. The plumes are of concern to the local people who rely on groundwater as a drinking water source. We used the freshwater turtle as a sentinel species to monitor the reproductive effects of exposure and, by inference, the potential for impact on human health. Our observations of male Chrysemys picta field-trapped from Moody Pond (an impacted site) and Washburn Pond (a reference site) on Cape Cod extended and supported prior observations of reproductive deficits. Morphometric comparison of precloacal length (PCL), which is a sexually dimorphic trait in the turtle, showed that Moody Pond males had a significantly longer PCL than Washburn Pond males. Moody Pond turtles showed reduced testicular weight, which was associated with significantly smaller seminiferous tubule diameter. Epididymal sperm counts were also markedly reduced in Moody Pond animals compared to Washburn Pond animals. Testicular histology and gonial proliferation, as determined by PCNA, were similar in both male populations, while the Moody Pond males had significantly higher germ cell apoptosis than the animals in Washburn Pond. These results suggest that a low level mixture of xenobiotic contaminants impairs the reproductive functions of turtles exposed to the impacted site but not to the reference site environment. PMID- 17123947 TI - Nurses' use and understanding of evidence-based practice: some preliminary evidence. PMID- 17123948 TI - Role of exogenous arginine vasopressin in the management of catecholamine refractory septic shock. PMID- 17123949 TI - Heparin-induced thrombocytopenia: advances in diagnosis and treatment. PMID- 17123950 TI - Decreasing vascular complications after percutaneous coronary interventions: partnering to improve outcomes. PMID- 17123951 TI - Certification: a benchmark for critical care nursing excellence. PMID- 17123952 TI - An acute care nurse practitioner model of care for neurosurgical patients. PMID- 17123953 TI - Ask the experts. Idiopathic pulmonary fibrosis. PMID- 17123955 TI - Conversion of pre-RISC to holo-RISC by Ago2 during assembly of RNAi complexes. AB - In the Drosophila RNA interference (RNAi) pathway, small interfering RNAs (siRNAs) direct Argonaute2 (Ago2), an endonuclease, within the RNA-induced silencing complex (RISC) to cleave complementary mRNA targets. In vitro studies have shown that, for each siRNA duplex, RISC retains only one strand, the guide, and releases the other, the passenger, to form a holo-RISC complex. Here, we have isolated a new Ago2 mutant allele and provide, for the first time, in vivo evidence that endogenous Ago2 slicer activity is important to mount an RNAi response in Drosophila. We demonstrate in vivo that efficient removal of the passenger strand from RISC requires the cleavage activity of Ago2. We have also identified a new intermediate complex in the RISC assembly pathway, pre-RISC, in which Ago2 is stably bound to double-stranded siRNA. PMID- 17123956 TI - KISS: the kinetoplastid RNA editing sequence search tool. AB - Kinetoplastid mitochondrial mRNA editing is a post-transcriptional process of uridine insertion and deletion. Editing is mediated by small RNA molecules termed guide RNAs (gRNAs). Most gRNAs are encoded by numerous small circular DNA minicircles, while the protein coding mitochondrial genes are encoded on a separate, larger genome called the maxicircle. In order to provide a workbench for the analysis of RNA editing in kinetoplastids and a well-annotated set of guide RNAs for Trypanosoma brucei, we generated the kinetoplastid RNA editing sequence search tool (KISS) (http://gmod.mbl.edu/kiss/). KISS is a pipeline and database that uses BLAST comparisons and minicircle sequence motifs to annotate potential gRNAs and cognate mRNA editing sequence. KISS 1.0 contains all previously known minicircle and maxicircle data from Trypanosoma brucei plus >400 new minicircle sequences. Using an online format, KISS 1.0 allows the mapping and visualization of all known T. brucei gRNAs to minicircle genes and to potential mRNA substrates for RNA editing. PMID- 17123957 TI - Vaccinia virus N1L protein resembles a B cell lymphoma-2 (Bcl-2) family protein. AB - Poxviruses encode immuno-modulatory proteins capable of subverting host defenses. The poxvirus vaccinia expresses a small 14-kDa protein, N1L, that is critical for virulence. We report the crystal structure of N1L, which reveals an unexpected but striking resemblance to host apoptotic regulators of the B cell lymphoma-2 (Bcl-2) family. Although N1L lacks detectable Bcl-2 homology (BH) motifs at the sequence level, we show that N1L binds with high affinity to the BH3 peptides of pro-apoptotic Bcl-2 family proteins in vitro, consistent with a role for N1L in modulating host antiviral defenses. PMID- 17123958 TI - The effects of cosolutes on protein dynamics: the reversal of denaturant-induced protein fluctuations by trimethylamine N-oxide. AB - The protein stabilizing effects of the small molecule osmolyte, trimethylamine N oxide, against chemical denaturant was investigated by NMR spin-relaxation measurements and model-free analysis. In the presence of 0.7 M guanidine hydrochloride increased picosecond-nanosecond dynamics are observed in the protein ribonuclease A. These increased fluctuations occur throughout the protein, but the most significant increases in flexibility occur at positions believed to be the first to unfold. Addition of 0.35 M trimethylamine N-oxide to this destabilized form of ribonuclease results in significant rigidification of the protein backbone as assessed by (1)H-(15)N order parameters. Statistically, these order parameters are the same as those measured in native ribonuclease indicating that TMAO reduces the amplitude of backbone fluctuations in a destabilized protein. These data suggest that TMAO restricts the bond vector motions on the protein energy landscape to resemble those motions that occur in the native protein and points to a relation between stability and dynamics in this enzyme. PMID- 17123959 TI - BPPred: a Web-based computational tool for predicting biophysical parameters of proteins. AB - We exploit the availability of recent experimental data on a variety of proteins to develop a Web-based prediction algorithm (BPPred) to calculate several biophysical parameters commonly used to describe the folding process. These parameters include the equilibrium m-values, the length of proteins, and the changes upon unfolding in the solvent-accessible surface area, in the heat capacity, and in the radius of gyration. We also show that the knowledge of any one of these quantities allows an estimate of the others to be obtained, and describe the confidence limits with which these estimations can be made. Furthermore, we discuss how the kinetic m-values, or the Beta Tanford values, may provide an estimate of the solvent-accessible surface area and the radius of gyration of the transition state for protein folding. Taken together, these results suggest that BPPred should represent a valuable tool for interpreting experimental measurements, as well as the results of molecular dynamics simulations. PMID- 17123960 TI - Solution structure of a small protein containing a fluorinated side chain in the core. AB - We report the first high-resolution structure for a protein containing a fluorinated side chain. Recently we carried out a systematic evaluation of phenylalanine to pentafluorophenylalanine (Phe --> F(5)-Phe) mutants for the 35 residue chicken villin headpiece subdomain (c-VHP), the hydrophobic core of which features a cluster of three Phe side chains (residues 6, 10, and 17). Phe --> F(5)-Phe mutations are interesting because aryl-perfluoroaryl interactions of optimal geometry are intrinsically more favorable than either aryl-aryl or perfluoroaryl-perfluoroaryl interactions, and because perfluoroaryl units are more hydrophobic than are analogous aryl units. Only one mutation, Phe10 --> F(5) Phe, was found to provide enhanced tertiary structural stability relative to the native core (by approximately 1 kcal/mol, according to guanidinium chloride denaturation studies). The NMR structure of this mutant, described here, reveals very little variation in backbone conformation or side chain packing relative to the wild type. Thus, although Phe --> F(5)-Phe mutations offer the possibility of greater tertiary structural stability from side chain-side chain attraction and/or side chain desolvation, the constraints associated with the native c-VHP fold apparently prevent the modified polypeptide from taking advantage of this possibility. Our findings are important because they complement several studies that have shown that fluorination of saturated side chain carbon atoms can provide enhanced conformational stability. PMID- 17123961 TI - Efficient molecular docking of NMR structures: application to HIV-1 protease. AB - Docking ligands into an ensemble of NMR conformers is essential to structure based drug discovery if only NMR structures are available for the target. However, sequentially docking ligands into each NMR conformer through standard single-receptor-structure docking, referred to as sequential docking, is computationally expensive for large-scale database screening because of the large number of NMR conformers involved. Recently, we developed an efficient ensemble docking algorithm to consider protein structural variations in ligand binding. The algorithm simultaneously docks ligands into an ensemble of protein structures and achieves comparable performance to sequential docking without significant increase in computational time over single-structure docking. Here, we applied this algorithm to docking with NMR structures. The HIV-1 protease was used for validation in terms of docking accuracy and virtual screening. Ensemble docking of the NMR structures identified 91% of the known inhibitors under the criterion of RMSD < 2.0 A for the best-scored conformation, higher than the average success rate of single docking of individual crystal structures (66%). In the virtual screening test, on average, ensemble docking of the NMR structures obtained higher enrichments than single-structure docking of the crystal structures. In contrast, docking of either the NMR minimized average structure or a single NMR conformer performed less satisfactorily on both binding mode prediction and virtual screening, indicating that a single NMR structure may not be suitable for docking calculations. The success of ensemble docking of the NMR structures suggests an efficient alternative method for standard single docking of crystal structures and for considering protein flexibility. PMID- 17123962 TI - Direct detection of transient alpha-helical states in islet amyloid polypeptide. AB - The protein islet amyloid polypeptide (IAPP) is a glucose metabolism associated hormone cosecreted with insulin by the beta-cells of the pancreas. In humans with type 2 diabetes, IAPP deposits as amyloid fibers. The assembly intermediates of this process are associated with beta-cell death. Here, we examine the rat IAPP sequence variant under physiological solution conditions. Rat IAPP is mechanistically informative for fibrillogenesis, as it samples intermediate-like states but does not progress to form amyloid. A central challenge was the development of a bacterial expression system to generate isotopically labeled IAPP without terminal tags, but which does include a eukaryotic post translational modification. While optical spectroscopy shows IAPP to be natively unfolded, NMR chemical shifts of backbone and beta-carbon resonances reveal the sampling of alpha-helical states across a continuous stretch comprising approximately 40% of the protein. In addition, the manifestation of nonrandom coil chemical shifts is confirmed by the relative insensitivity of the amide proton chemical shifts to alterations in temperature. Intriguingly, the residues displaying helical propensity are conserved with the human sequence, suggesting a functional role for this conformational bias. The inability of rat IAPP to self assemble can be ascribed, in part, to several slowly exchanging conformations evident as multiple chemical shift assignments in the immediate vicinity of three proline residues residing outside of this helical region. PMID- 17123964 TI - Infection by human immunodeficiency virus-1 is not a risk factor for amebiasis. PMID- 17123965 TI - Molecular epidemiology of malaria in Cameroon. XXIII. Experimental studies on serum substitutes and alternative culture media for in vitro drug sensitivity assays using clinical isolates of Plasmodium falciparum. AB - Correlation studies on the in vitro drug response of field isolates of Plasmodium falciparum and molecular markers for drug resistance are becoming important as many malaria control programs abandon monotherapies and resort to combination therapies. The standardization and optimization of the in vitro drug sensitivity assay are one of the prerequisites for validating molecular markers in the field. The present study was designed to assess and compare the growth of freshly obtained isolates for at least the first erythrocytic cycle in various culture media and determine the in vitro response to chloroquine in alternative media. Parasite growth was consistently higher in Dulbecco's modified Eagle's medium (DME)-human serum, Iscove's modified Dulbecco's medium (IMDM)-human serum, RPMI 1640 medium-goat serum, and a serum-free medium containing 1:1 (v/v) mixture of IMDM and F-12 supplemented with an ammonium sulfate fraction of adult bovine serum than in RPMI 1640 medium-human serum mixture. The level of chloroquine response determined in human serum-supplemented DME, IMDM, and RPMI 1640 media did not differ significantly (P > 0.05) from the control (RPMI 1640-human serum). This study suggests that alternative media may be used to optimize parasite growth during the critical initial phase of transition from in vivo to in vitro conditions. The capacity of these media to support long-term cultivation of P. falciparum requires further investigation. PMID- 17123966 TI - Survey of community knowledge, attitudes, and practices during a malaria epidemic in central Java, Indonesia. AB - We surveyed adults in a randomly selected sample of 1,000 households in 50 villages in nine malarial sub-districts in Purworejo, central Java, Indonesia from May to July 2001. The survey assessed malaria knowledge, attitudes, and practices in communities experiencing epidemic malaria to begin exploring broad strategies for controlling the disease in the region. A pre-tested survey instrument consisting of 93 questions addressed demographic characteristics, socioeconomic factors, knowledge and perceptions of malaria, burden and severity of disease, treatment-seeking behavior, malaria prevention practices, and perceptions of government malaria control efforts. The survey was taken by in person interview of all subjects. Most (97%) subjects were aware of malaria and more than two-thirds correctly identified mosquitoes as the vector. Forty-one percent of households in both forest/hilly and agricultural/urban areas reported malaria illness in the past year. Thirty-six percent (357 households) owned at least one bed net, 92% of these had been purchased by the owners. However, only 36% of households with bed nets affirmed their use as a means of preventing malaria. Nearly all respondents reported a willingness to accept spraying of residual insecticides for malaria prevention, yet less than 5% were willing to pay a nominal fee (US $3) for this service. Fifty-two percent of respondents reported self-treatment of malaria illness without visiting a health facility. This assessment of knowledge, attitudes, and practices showed a broad awareness of malaria and its consequences among residents of malarial areas in the Menoreh Hills of Central Java. PMID- 17123967 TI - Malarial retinopathy: a newly established diagnostic sign in severe malaria. AB - Severe malaria is commonly misdiagnosed in Africa, leading to a failure to treat other life-threatening illnesses. In malaria-endemic areas, parasitemia does not ensure a diagnosis of severe malaria because parasitemia can be incidental to other concurrent disease. The detection of malarial retinopathy is a candidate diagnostic test for cerebral malaria. Malarial retinopathy consists of a set of retinal abnormalities that is unique to severe malaria and common in children with cerebral malaria. Its presence and severity are related to risk of death and length of coma in survivors. A large, prospective autopsy study of children dying with cerebral malaria in Malawi found that malarial retinopathy was better than any other clinical or laboratory feature in distinguishing malarial from non malarial coma. However, visualization has to date relied on specialist examination techniques. Further studies are planned to evaluate the usefulness of funduscopy by general clinicians in a variety of settings across Africa. Studies of the retina and retinal blood vessels provide an unparalleled opportunity to visualize an infected microvasculature and its effect on neural tissue in vivo. This report reviews current knowledge of malarial retinopathy, including its use as a diagnostic test in the comatose child, and its value as a tool for research into the pathophysiology of cerebral malaria. PMID- 17123968 TI - Microscopic and sub-microscopic Plasmodium falciparum infection, but not inflammation caused by infection, is associated with low birth weight. AB - Pregnancy-associated malaria is one of the leading causes of low birth weight in malaria endemic areas. In this study, 145 parturient women residing in areas endemic for Plasmodium falciparum in Lambarene, Gabon, were recruited into the study after delivery, and the association of maternal P. falciparum infection, inflammatory response, and birth weight was studied. At delivery, 10% (15) of the mothers (12 were positive in both peripheral and placental blood smears, 1 was positive in peripheral blood only, and 2 were positive in placenta blood only) were positive for P. falciparum by microscopy and 23% (30) by real-time polymerase chain reaction (PCR). The level of C-reactive protein (CRP) was significantly elevated in microscopically P. falciparum-positive pregnant women (34 mg/L; 95% CI: 3-458) but not in those with sub-microscopic infections (6 mg/L; 95% CI: 1-40) compared with those free of P. falciparum infection (7 mg/L; 95% CI: 1-43). In a multivariate analysis, the presence of microscopic (adjusted OR = 28.6, 95% CI = 4.8-169.0) or sub-microscopic (adjusted OR = 13.2, 95% CI = 2.4-73.0) P. falciparum infection in pregnant women and age of mothers < 21 years (adjusted OR = 9.7 CI = 1.0-89.7), but not CRP levels, were independent predictors for low birth weight. This finding may have important operational implications and emphasizes the need for appropriate diagnostic methods in studies evaluating the outcome of pregnancy-associated malaria. PMID- 17123969 TI - Characterization of counterfeit artesunate antimalarial tablets from southeast Asia. AB - In southeast Asia, the widespread high prevalence of counterfeits tablets of the vital antimalarial artesunate is of great public health concern. To assess the seriousness of this problem, we quantified the amount of active ingredient present in artesunate tablets by liquid chromatography coupled to mass spectrometry. This method, in conjunction with analysis of the packaging, classified tablets as genuine, substandard, or fake and validated results of the colorimetric Fast Red TR test. Eight (35%) of 23 fake artesunate samples contained the wrong active ingredients, which were identified as different erythromycins and paracetamol. Raman spectroscopy identified calcium carbonate as an excipient in 9 (39%) of 23 fake samples. Multivariate unsupervised pattern recognition results indicated two major clusters of artesunate counterfeits, those with counterfeit foil stickers and containing calcium carbonate, erythromycin, and paracetamol, and those with counterfeit holograms and containing starch but without evidence of erythromycin or paracetamol. PMID- 17123970 TI - Epidemiology of malaria in an area of low transmission in central India. AB - A longitudinal study on malaria was carried out from 2003 to 2005 in an area of unstable malaria in the Panna district in central India. Both Plasmodium vivax and P. falciparum were prevalent; however, the risk of P. falciparum malaria was 31.6% (95% confidence interval [CI] = 29.6-33.6%), which is four times higher compared with that of P. vivax malaria (7.8%, 95% CI = 6.7-9%). An increasing trend was recorded in malaria prevalence from 30.2% in 2003 to 46.6% in 2004 (odds ratio [OR] = 2.0, 95% CI = 1.6-2.5) that increased to 58.6% in 2005 (OR = 1.6, 95% CI = 1.2-2.1). This increase was statistically significant (chi(2) = 120.5, degrees of freedom = 2, P < 0.0001). Anopheles culicifacies was the dominant vector of malaria and showed partial (< 50%) resistance to DDT, which indicated that DDT can still be used. Improved access to treatment facilities, combination therapy, and vector control appears to be the most promising method for controlling malaria in this region. PMID- 17123971 TI - The evaluation of radiolabeled artesunate on tissue distribution in rats and protein binding in humans. AB - The present study reports the tissue distribution, pharmacokinetics, mass balance, and elimination of [(14)C] artesunate (AS) following single intravenous administration in rats. Protein binding was performed with rat and human plasma. Radioactivity and drug levels in blood, plasma, tissues, urine, and feces up to 192 hours were collected and measured. The mean terminal half-life of plasma (76 h) and blood (105 h) radioactivity was prolonged compared with that of unchanged AS (0.43 h) and dihydroartemisinin (0.75 h), an active metabolite of AS. Drug was widely distributed after 1 hour in select tissues. After 24 hours, the radioactivity rapidly declined in all tissues except spleen until 96 hours. Only 1% of total radioactivity was detected in brain tissue. AS revealed a higher binding capacity with human and rat plasma proteins (73-81%). The radioactivity in whole blood was higher (two to fourfold) than that in plasma throughout the period of the treatment, suggesting that AS binding to RBCs may relate to its powerful antimalarial activity. PMID- 17123972 TI - Short report: Severe Plasmodium falciparum malaria in Cameroon: associated with the glutathione S-transferase M1 null genotype. AB - Glutathione S-transferases (GST) are a family of enzymes involved in phase-II detoxification of endogenous and xenobiotic compounds. Polymorphisms in GST genes have been associated with susceptibility to different diseases. In this study we determined the frequencies of polymorphisms in GSTM1, GSTT1, and GSTP1 in DNA of 138 children from Cameroon, presenting with uncomplicated malaria (N = 19), malaria with minor complications (N = 81), or severe malaria (N = 38). Analyses of GSTM1 and GSTT1 were performed using PCR-multiplex procedure, while GSTP1 was done by PCR-RFLP. Subjects presenting with malaria with complications were found more often of the GSTM1-null genotype (58-64%) as compared with those with uncomplicated malaria (32%), a difference that was statistically significant. We conclude that the GSTM1-null genotype is associated with malaria with complications. PMID- 17123973 TI - Genetic diversity of Plasmodium vivax Pvcsp and Pvmsp1 in Guyana, South America. AB - Approximately 55% of malaria infections in the Guyana Amazon region are attributed to Plasmodium falciparum while the other 45% are attributed to non falciparum, mostly Plasmodium vivax. However, little is known about the P. vivax strain types circulating in the region. Using PCR for Plasmodium detection and two genetic markers specific to P. vivax to detect the polymorphic circumsporozoite protein (CSP) and the conserved 19-kDa region of the merozoite surface protein-1 (MSP-1), we investigated the overall Plasmodium strain distribution and population diversity within P. vivax in isolates collected from the blood of infected individuals in the interior Amazon region of Guyana, South America. Out of a total of 250 samples positive for Plasmodium, P. vivax was detected in 30% (76/250) and P. falciparum was detected in 76% (189/250). Mixed infections containing both P. falciparum and P. vivax constituted 6% (15/250) of the total positive samples. Further analysis of P. vivax strains showed that 92% (56/61) of the P. vivax samples hybridized with a probe specific to type VK210, 39% (24/61) hybridized with a probe specific for type VK247, and 25% (15/61) hybridized with a probe specific for the P. vivax-like CS genotype. DNA sequencing of the 19-kDa C-terminal domain in block 13 of MSP-1 amplified from 61 samples from patients infected with P. vivax demonstrated that this region is highly conserved, and all samples were identical at the nucleotide level to the Belem and Salvador-1 types. No synonymous or nonsynonymous mutations were observed in this region of the gene, indicating that current vaccine-development efforts based on the MSP-1(19) fragment would be applicable in Guyana. PMID- 17123974 TI - Validation of microsatellite markers for use in genotyping polyclonal Plasmodium falciparum infections. AB - Genotyping methods for Plasmodium falciparum drug efficacy trials have not been standardized and may fail to accurately distinguish recrudescence from new infection, especially in high transmission areas where polyclonal infections are common. We developed a simple method for genotyping using previously identified microsatellites and capillary electrophoresis, validated this method using mixtures of laboratory clones, and applied the method to field samples. Two microsatellite markers produced accurate results for single-clone but not polyclonal samples. Four other microsatellite markers were as sensitive as, and more specific than, commonly used genotyping techniques based on merozoite surface proteins 1 and 2. When applied to samples from 15 patients in Burkina Faso with recurrent parasitemia after treatment with sulphadoxine-pyrimethamine, the addition of these four microsatellite markers to msp1 and msp2 genotyping resulted in a reclassification of outcomes that strengthened the association between dhfr 59R, an anti-folate resistance mutation, and recrudescence (P = 0.31 versus P = 0.03). Four microsatellite markers performed well on polyclonal samples and may provide a valuable addition to genotyping for clinical drug efficacy studies in high transmission areas. PMID- 17123975 TI - Identification of a novel gene encoding a secreted antigen 1 of Babesia gibsoni and evaluation of its use in serodiagnosis. AB - Serum from a dog immunized with blood plasma from a B. gibsoni-infected dog, putatively containing secreted antigens, was used to screen a cDNA expression library. A novel gene encoding BgSA1 was identified from the isolated clones. The serum raised in mice immunized with the recombinant BgSA1 expressed in Escherichia coli could recognize a native parasite protein with a molecular mass of 59 kDa. Comparing with the previously established ELISA with recombinant P50 as antigen, the ELISA with recombinant BgSA1 as the antigen was more sensitive when they were used to detect field samples. Moreover, a sandwich ELISA with anti BgSA1 antibodies could detect the circulating BgSA1 in a serial blood plasma from a dog experimentally infected with B. gibsoni. These results indicated that BgSA1 could be a useful target for the development of a diagnostic test for the detection of specific antibodies and circulating antigens. PMID- 17123976 TI - Cryptosporidium hominis: experimental challenge of healthy adults. AB - Cryptosporidium hominis causes diarrhea in humans and has been associated with community outbreaks. This study describes the infectivity, illness, and serologic response after experimental challenge of 21 healthy adult volunteers with 10-500 C. hominis (TU502) oocysts. Sixteen subjects (76.2%) had evidence of infection; the 50% infectious dose (ID(50)) was estimated to be 10-83 oocysts using clinical and microbiologic definitions of infection, respectively. Diarrhea occurred in 40% of subjects receiving 10 oocysts with a stepwise increase to 75% in those receiving 500 oocysts. A serum IgG response was seen in those receiving more than 30 oocysts. Greatest responses were seen in volunteers with diarrhea and oocyst shedding. Volunteers with no evidence of infection had indeterminant or negative IgG responses. Cryptosporidium hominis 10 oocysts) and is clinically is infectious for healthy adults (ID(50) = similar to C. parvum-induced illness. In contrast to C. parvum, C. hominis elicted a serum IgG response in most infected persons. PMID- 17123977 TI - Reference values for Leishmania infantum parasitemia in different clinical presentations: quantitative polymerase chain reaction for therapeutic monitoring and patient follow-up. AB - Quantification of Leishmania infantum DNA in blood samples by an ultrasensitive quantitative polymerase chain reaction (QPCR) detected parasitemias in different clinical presentations. We observed a large range of parasitemias, more than 9 log values, and could determine the threshold between asymptomatic carriage and disease in the Mediterranean area (approximately one parasite/mL of blood). Based on kinetoplast DNA amplification, this assay had a sensitivity of 0.001 parasite DNA equivalents/mL and detected asymptomatic carriage of Leishmania. It detected parasite DNA in 58% of healthy subjects, while an immunoblot detected specific antibodies in only 16%. For initial diagnosis of disease, this quantitative PCR with blood samples constitutes a non-invasive alternative to bone marrow aspiration. Its main applications are monitoring of drug therapy and follow-up of immunodeficient patients for biologic confirmation of relapses. PMID- 17123978 TI - Cystatin C as a marker of immune complex-associated renal impairment in a Sudanese population with visceral leishmaniasis. AB - Renal function was studied in visceral leishmaniasis (VL) and post-kala-azar dermal leishmaniasis (PKDL) by means of the specific marker cystatin C and related to circulating immune complexes and cytokine production. Forty patients with VL (23 with sub-acute disease and 17 with acute disease), 17 patients with PKDL, and 22 healthy controls were included. Cystatin C, but not creatinine, was significantly raised in VL (P = 0.004). The highest levels of cystatin C were found in those with acute disease (P < 0.0001). In VL, cystatin C levels were positively correlated to circulating immune complexes and production of granulocyte-macrophage colony stimulating factor (GM-CSF), but negatively correlated to aspartate aminotransferase and lactate dehydrogenase. We conclude that cystatin C is a superior marker of glomerular function in leishmaniasis and that immune complex deposition and GM-CSF are two functions that most likely are causally involved in the mechanisms leading to glomerular dysfunction in leishmaniasis. PMID- 17123979 TI - Short report: Human trypanosomiasis caused by Trypanosoma evansi in a village in India: preliminary serologic survey of the local population. AB - After discovery of the first recorded case of human infection with Trypanosoma evansi, serologic screening of 1,806 persons from the village of origin of the patient in India was performed using the card agglutination test for trypanosomiasis and T. evansi. A total of 410 (22.7%) people were positive by whole blood, but only 81 were confirmed positive by serum. However, no trypanosomes were detected in the blood of 60 people who were positive at a high serum dilution. The results probably indicate frequent exposure of the human population to T. evansi in the study area, which suggests frequent vector transmission of parasites to humans. Although T. evansi is not infective for humans, a follow-up of seropositive persons is required to observe the evolution of human infection with this parasite. PMID- 17123980 TI - Congenital Chagas disease in Bolivia is not associated with DNA polymorphism of Trypanosoma cruzi. AB - This study aims to typify the Trypanosoma cruzi (sub)lineage(s) in umbilical cord blood of congenitally infected Bolivian newborns, using PCR amplifications of "Region Markers", mini-exon or kDNA fragments followed by hybridization or sequencing. New probes were also designed to distinguish three variants within the TcIId sublineage. The IIb, IId, or IIe T. cruzi sublineages, as well as different variants of the IId sublineage, were detected in infected neonates, whereas mixed infections were not found. The frequencies of the IId sublineage were similar in neonates (95.1%) and adults of the same area (94.1%). The IId infected newborns displayed either asymptomatic, or severe and fatal clinical forms of congenital Chagas disease, as well as low or high parasitemia. Altogether these data show that T. cruzi DNA polymorphism, based on the presently available markers, is not associated with the occurrence of congenital infection or the development of severe clinical forms of congenital Chagas disease. PMID- 17123981 TI - Changes in bacterial profile during amebiasis: demonstration of anaerobic bacteria in ALA pus samples. AB - Little is known about the changes in gut resident flora during amebic colitis and amebic liver abscess (ALA) caused by Entamoeba histolytica infection. Fecal samples from ALA patients, from healthy E. histolytica negative and positive (asymptomatic) individuals, and from pre- and post-metronidazole-treated healthy volunteers and pus samples from ALA patients were tested for the presence of various bacterial genera using 16S rRNA-based primers. Statistically significant reduction in Lactobacillus due to E. histolytica infection was observed in asymptomatic individuals and ALA patients. On the other hand, reduction in Bacteroides, Bifidobacterium, and Clostridium in the same samples was due to metronidazole treatment. Two anaerobic genera, viz. Bacteroides and Peptostreptococcus, were detected in ALA pus samples, and this observation is unprecedented. In addition, PCR revealed metronidazole resistance genes in fecal and pus samples of metronidazole-treated individuals. Re-examination of the ameba bacterium relationship in amebiasis is suggested. PMID- 17123982 TI - Aedes aegypti vectorial capacity is determined by the infecting genotype of dengue virus. AB - Dengue viruses causing severe, hemorrhagic disease have displaced less virulent strains in the Americas during the past three decades. The American (AM) genotype of dengue serotype 2 has been endemic in the Western Hemisphere and South Pacific, causing outbreaks of dengue fever (DF), but has not been linked to dengue hemorrhagic fever (DHF). The Southeast Asian (SEA) genotype of dengue was introduced into this hemisphere in 1981, has caused outbreaks with numerous cases of DHF, and has displaced the AM genotype in several countries. We investigated the effect of viral genotype on the potential for transmission by infecting Aedes aegypti mosquitoes collected in South Texas with six viruses, representing these two genotypes. Viral replication in the midgut was significantly higher in SEA infected mosquitoes, and virus-specific proteins could be detected in salivary glands 7 days earlier in SEA- than AM-infected mosquitoes. This much earlier appearance of dengue virus in salivary glands resulted in an estimated 2- to 65 fold increase in the vectorial capacity of these mosquitoes for the viruses that can cause DHF. This may be one of the mechanisms through which more virulent flaviviruses spread and displace others globally. PMID- 17123983 TI - Triatoma infestans chooses to feed upon immune prey. AB - Blood-feeding Triatoma infestans obtained its fills from immune chickens in 15 min, but it needed 40 min for feeding upon non-immune chickens. High-titer specific IgGs and skin reactivity against T. infestans saliva antigens were elicited in immune chickens. Fluorescence-labeled leukocytes from non-immune or immune chickens were used to determine sources of blood drawn by equal numbers of triatomines distributed in separate compartments of a hut-like box. It was shown that 64.4 +/- 4.7% of the reduviids were captured in the immune chicken room; 35.6 +/- 4.5% were present in the non-immune chicken dwelling, and these differences were statistically significant (P < 0.001). Furthermore, T. infestans feeding upon immune birds reached the adult stage 40 days before those feeding upon non-immune birds, and differences were statistically significant. These results appear to have a broad epidemiologic significance as for spreading enzootics; hence, the immunologic status of vertebrate host populations appears to favor T. infestans as the main transmitter of Trypanosoma cruzi. PMID- 17123984 TI - Variation in recombination rate across the X chromosome of Anopheles gambiae. AB - The M and S molecular forms of Anopheles gambiae are considered to be incipient species, despite residual gene exchange. Of the three small genome regions that are strongly differentiated between the molecular forms ("speciation islands"), two are located near centromeres, on the left arm of chromosome 2 and the X chromosome. To test the prediction of reduced recombination in these islands, we estimated recombination rates between microsatellite loci on the X chromosome using two M-form strains. Across most of the chromosome, recombination occurred at approximately 1 centimorgan per megabase (cM Mb(-1)), a value closely matching the genome-wide average estimated for A. gambiae and for other eukaryotes. Recombination was much higher at the telomeric end, > 7 cM Mb(-1). In the speciation island at the centromeric end, recombination was sharply reduced to approximately 0.2 cM Mb(-1), consistent with a role for reduced recombination in maintaining differentiation between nascent species despite gene flow. PMID- 17123985 TI - Biochemical and immunologic predictors of efficacy of treatment or reinfection risk for Schistosoma mansoni. AB - Most Schistosoma mansoni infections are egg-negative after a single dose of oxamniquine. A cohort of 661 infected children was treated at 6-month intervals and assessed for nutritional and parasitological status. Initial biochemical and immunologic markers were measured in a subset of 84 children. All were treated at the start of therapy and at 6 months. Immunoglobulins only served as markers for active infection. No markers were predictive of cure or reinfection, except initial infection intensity and serum low-density lipoprotein. Ten percent were persistently infected and had no change in infection intensity at any time-point. Several factors suggest that this group was biologically different. In addition to failing to reduce their worm burden, they had significantly higher initial intensity of infection (100 versus 65 eggs/g, P = 0.001) and significantly lower initial serum low-density lipoprotein (72 versus 104 mg/dL, P = 0.045). The biologic plausibility of this observation is discussed. PMID- 17123986 TI - CD4+ T-cell counts, CD4+/CD8+ T-cell count ratios, and antibody levels in migrant fishermen infected with Schistosoma japonicum in the Dongting Lake, China. AB - In this study, CD4(+) and CD8(+) T cells and antibody levels were measured in 94 migrant fishermen infected with Schistosoma japonicum from Dongting Lake, China. Prevalence among these fishermen was high (63.8%), with a mean infection intensity of 61.4 +/- 3.8 epg, and included a high proportion of individuals (39.4%) with substantial parenchymal fibrosis (stages > or = 2/3). The CD4(+)/CD8(+) ratio in men (1.34 +/- 0.11) was significantly lower than that of women (1.58 +/- 0.15). CD4(+) T cells and the ratio of CD4(+)/CD8(+) were significantly decreased both in subjects infected with S. japonicum and those with parenchymal fibrosis. However, levels of total IgA, IgM, and anti schistosome egg antigen IgG correlated positively with infection intensity and pathologic lesion number. These results suggest an imbalance between cell mediated and humoral immunity in these fishermen, the precise cause of which remains undetermined. PMID- 17123987 TI - Dose-ranging study for trials of therapeutic infection with Necator americanus in humans. AB - Epidemiological studies suggest that a hookworm infection producing 50 eggs/gram of feces may protect against asthma. We conducted a dose-ranging study to identify the dose of hookworm larvae necessary to achieve 50 eggs/gram of feces for therapeutic trials of asthma. Ten healthy subjects without asthma or airway hyperresponsiveness to inhaled methacholine received 10, 25, 50, or 100 Necator americanus larvae administered double blind to an area of skin on the arm. Subjects were seen weekly for 12 weeks and were then treated with mebendazole. Skin itching at the entry site and gastrointestinal symptoms were common at higher doses. Lung function did not change. Levels of blood eosinophils and IgE increased transiently, and levels of IgG increased progressively. All doses resulted in at least 50 eggs/gram of feces in the eight subjects who completed the study. Infection with 10 N. americanus larvae is well tolerated, elicits a modest host eosinophil response, and is potentially suitable for use in preliminary clinical therapeutic trials. PMID- 17123988 TI - Modeling targeted ivermectin treatment for controlling river blindness. AB - There is considerable host heterogeneity in exposure to onchocerciasis. We incorporate this heterogeneity into a model of onchocerciasis transmission that we use to evaluate intervention strategies targeting specific portions of the human population for treatment with ivermectin. Our model predicts that targeted allocation of ivermectin in a highly heterogeneous population will reduce the public health burden of onchocerciasis using 20-25% of the doses of untargeted allocation. Targeted allocation therefore poses significantly lower risk of adverse effects, while potentially delaying the emergence and spread of ivermectin resistance, relative to untargeted allocation. PMID- 17123989 TI - Symptoms reported after mass drug administration for lymphatic filariasis in Leogane, Haiti. AB - Mass drug administration (MDA) for lymphatic filariasis (LF) can cause adverse reactions from microfilarial and adult worm death. Symptoms after the fifth annual MDA in Leogane, Haiti, were studied to determine whether they resulted from parasite death. Persons reporting post-MDA systemic symptoms at 5 of 148 drug distribution posts and men reporting scrotal pain at any post were interviewed. Participants were tested with immunochromatographic tests (ICTs), and men with scrotal symptoms were examined. At the five posts, 3,781 persons took anti-filarial medication. Of these, 314 (8%) returned with symptoms; the most common were headache (36%) and gastrointestinal complaints (28%). Of the 294 (94%) who consented to ICT testing, 47 (16%) were positive. Of 69 men with scrotal symptoms who consented to ICT testing, 18 (26.1%) were positive. After Leogane's fifth MDA, most symptomatic persons had undetectable levels of filarial antigen by ICT. Free symptomatic treatment may motivate some people to report symptoms and seek care. PMID- 17123990 TI - Short report: Evidence and potential for transmission of human and swine Taenia solium cysticercosis in the Piracuruca region, Piaui, Brazil. AB - The study conducted in the Cocal dos Alves municipality, located in the Piracuruca region of Piaui State, Brazil in November of 2003 was based on both a socio-behavioral survey and analysis of serum antibodies and parasitic materials. Pig raising is the main economic activity with 91.4% using extensive farming. On the studied population, 54.3% of people did not apply any sanitary measures to wastewater, 45.7% used septic tanks, and 69.1% consumed water without treatment. The data collected indicated the occurrence and active transmission of the taeniasis-cysticercosis complex in the region. One of seven voluntary persons was positive in antibody-ELISA tests using both native and recombinant antigens. Multiplex PCR and DNA sequencing of cyst samples obtained from a pig revealed the presence of the Afro-American genotype of Taenia solium. This study revealed the occurrence of human and porcine cysticercosis in the Piracuruca region of Piaui State, middle-north of Brazil. PMID- 17123991 TI - Mucocutaneous entomophthoramycosis acquired by conjunctival inoculation of the fungus. AB - Entomophthoramycoses are classified into subcutaneous, mucocutaneous, and primary visceral forms. The mucocutaneous form, also known as rhinoentomophthoramycosis, involves the mucosa and subcutaneous tissues of the nose and is caused by Conidiobolus coronatus (Entomophthora coronata). In this report, we describe the first case of mucocutaneous entomophthoramycosis acquired by introduction of the fungus through the conjunctival mucosa as a consequence of trauma involving contamination with soil. The patient was a 37-year-old man with no other complaints. The lesion was tumoral and extended into the ethmoidal and maxillary sinuses. The histopathologic appearance of the lesion was characteristic of this infection with a granulomatous process rich in eosinophils and with hyphae surrounded by an eosinophilic, periodic acid-Schiff stain-positive halo (Splendore-Hoeppli phenomenon). To the best of our knowledge, this case constitutes the first report of mucocutaneous entomophthoramycosis acquired by ocular contamination. PMID- 17123992 TI - Moderate- and low-intensity co-infections by intestinal helminths and Schistosoma mansoni, dietary iron intake, and anemia in Brazilian children. AB - To determine the role of moderate- and low-intensity infections with Schistosoma mansoni and intestinal helminths (hookworm, Trichuris trichiura, Ascaris lumbricoides) on the prevalence of anemia and their relationship to iron consumption, a cross-section of 1,709 children in rural Brazil was studied. All participants were selected for infection with one or multiple helminthic parasites, and demographic, anthropometric, and dietary intake were surveyed. The prevalence and intensity were as follows: hookworm infection, 15.7% and 8.6 eggs/g; T. trichiura, 74.8% and 190.5 eggs/g; A. lumbricoides, 63% and 1,905.5 eggs/g; S. mansoni, 44.5% and 60.3 eggs/g. There was no increase in odds ratio for anemia with any combination of intestinal helminths without S. mansoni infection. By logistic regression, the odds ratio for having anemia when infected with S. mansoni and two intestinal helminths was 1.7 (95% CI, 1.1-2.5) and for S. mansoni and three intestinal helminths was 2.4 (95% CI, 1.2-4.6) compared with children with a single parasite species. Children with an adequate intake of iron had no increased odds of anemia independent of the combination of parasite infections. PMID- 17123993 TI - Short report: Melioidosis in Myanmar: forgotten but not gone? AB - A serologic survey of adults resident in Myanmar was conducted to define the presence of antibodies to Burkholderia pseudomallei, the cause of melioidosis. Antibodies were detectable by indirect hemagglutination assay (IHA) in 757 (78%) of 968 adults, of whom 69 (7%) had an IHA titer > or =1:160. PMID- 17123994 TI - Melioidosis in the Americas. AB - Melioidosis is a potentially severe bacterial infection caused by Burkholderia pseudomallei. There has been growing awareness of the disease in the Americas, particularly since the Vietnam conflict when it was diagnosed in returning service personnel. Accidental laboratory exposure indicates the difficulty making a culture-based diagnosis when melioidosis has not been considered in the differential diagnosis. Melioidosis is most likely underdiagnosed in tropical Central and South America where conditions are more suited to persistence of B. pseudomallei in the environment. Recent melioidosis case clusters in northeastern Brazil highlight the threat posed to rural populations located far from specialist services. Increased clinical awareness of the disease and improvements in laboratory diagnostic methods are likely to bring wider recognition of melioidosis in the Americas. PMID- 17123995 TI - Scrub typhus during pregnancy and its treatment: a case series and review of the literature. AB - Although scrub typhus is uncommon in pregnant women, when present, it can have serious repercussions for the mother and developing fetus. Since it is uncommon, the clinical impact of scrub typhus on pregnancy has not been elucidated and an effective and safe therapeutic regimen has not been validated. The medical records of pregnant women whose scrub typhus were treated at Chungnam National University Hospital were reviewed and their clinical outcomes were evaluated. A review of the literature was also performed on pregnant women with scrub typhus and their clinical outcomes. Eight pregnant women with scrub typhus were treated successfully with a single 500-mg dose of azithromycin, and no relapses were reported. They all delivered healthy babies at term, without congenital or neonatal complications. In the reviews, azithromycin was effective against scrub typhus and had favorable pregnancy outcomes. Ciprofloxacin and cefuroxime failed to treat scrub typhus and fetal loss resulted. A single 500-mg dose of azithromycin may be a reasonable treatment regimen for pregnant women with scrub typhus. Ciprofloxacin might not be advisable for the treatment of scrub typhus during pregnancy. Scrub typhus itself seems to have serious adverse effects on pregnancy if not appropriately controlled. PMID- 17123996 TI - Utility of polymerase chain reaction in diagnosis of tuberculosis from samples of bone marrow aspirate. AB - Fever of unknown origin (FUO) poses a diagnostic challenge to the clinicians, with a differential diagnosis as varied as neoplastic and infectious diseases. In developing countries, the infectious causes are responsible for more cases of FUO, with tuberculosis as one of the main causes of classic FUO. Disseminated tuberculosis with negative pulmonary findings is a diagnostic problem. This study examines the diagnostic utility of the polymerase chain reaction (PCR) in samples of bone marrow aspirate in 85 patients presenting with diverse clinical symptoms. Using primers specific for Mycobacterium tuberculosis, tubercular etiology was detected in 33% of patients clinically suspected of tuberculosis while culture on Lowenstein-Jensen medium grew M. tuberculosis in only one patient (2.5%). None of these patients had been diagnosed by microscopy. Clinical improvement with ATT was observed in 85% of the patients with positive PCR. PCR demonstrated much higher sensitivity and specificity, thereby facilitating early therapeutic decisions for suspected extrapulmonary tuberculosis. PMID- 17123997 TI - Short report: Association of macrophage inflammatory response and cell death after in vitro Borrelia burgdorferi infection with arthritis resistance. AB - Susceptibility to Borrelia burgdorferi infection and subsequent arthritis is genetically determined in mice and determined by innate immunity. Accordingly, macrophage responses to B. burgdorferi challenge may differ between mouse strains. Bone marrow-derived macrophages were infected ex vivo with clonal B. burgdorferi strain N40. Interleukin-12 and tumor necrosis factor-alpha (TNF alpha) production were higher in macrophages from resistant C57Bl/6 mice than in macrophages from susceptible C3H/HeJ mice. However, TNF-alpha production was observed in lower concentrations in C3H/HeJ (toll-like receptor-4(-/-)) macrophages than in C3H/FeJ (TLR4(+/+)) macrophages, suggesting that TLR4 might contribute to the response to B. burgdorferi. A higher cytokine response to B. burgdorferi was associated with cell death in macrophages from resistant C57Bl/6 mice. Understanding variability in the response of macrophages to B. burgdorferi may contribute to understanding Lyme arthritis. PMID- 17123998 TI - Risk factors associated with diarrhea among international visitors to Cuzco, Peru. AB - The objective of this study was to determine the risk factors for travelers' diarrhea (TD) reported by visitors to Cuzco, Peru. In this cross-sectional study, self-administered questionnaires looking at perceived occurrence of health problems and pre-travel health advice were obtained from departing travelers at Cuzco's International Airport between August and November 2002. A total of 5,988 travelers participated in the study. The mean age was 35 years, and 51% were women. The prevalence of TD was 24%. Factors associated with TD in the multivariable analysis were use of antibiotic prophylaxis (OR: 3.20), vaccination against cholera (OR: 1.44), history of advice for safe food and water consumption (OR: 1.46), being younger than 35 years of age (OR: 1.37), being a resident of the United States (OR: 1.28), not staying in hotels in Cuzco (OR: 1.13), and number of unsafe food or beverages consumed (OR: 1.04). Remaining in Cuzco < 1 week was a protective factor (OR: 0.58). TD is common among travelers to Cuzco. This study suggests that conventional recommendations are not invariably effective at decreasing TD. PMID- 17123999 TI - Review of reported cholera outbreaks worldwide, 1995-2005. AB - The global temporal and spatial distribution of cholera is underappreciated, given the lack of surveillance in endemic areas and economic disincentives to report outbreaks. To judge the use of specific novel interventions such as vaccines or anti-secretory agents, we compiled a database and analyzed cholera reports from the Program for Monitoring Emerging Diseases from 1995 to 2005. Of the 632 reports meeting the search criteria, 66% originated in Sub-Saharan Africa, followed by 16.8% from Southeast Asia. Reported outbreaks in Africa tended to be larger in size. The most common risk factors were water source contamination, heavy rainfall and flooding, and population dislocation. While cholera reporting is sub-optimal, this review provides a detailed sub-national quantification of cholera, identifies foci of endemicity in Africa, and describes risk factors by region. We highlight the need for more extensive outbreak reporting to justify investments in new interventions. PMID- 17124000 TI - Causes of community-acquired bacteremia and patterns of antimicrobial resistance in Vientiane, Laos. AB - There is no published information on the causes of bacteremia in the Lao PDR (Laos). Between 2000 and 2004, 4512 blood culture pairs were taken from patients admitted to Mahosot Hospital, Vientiane, Laos, with suspected community-acquired bacteremia; 483 (10.7%) cultures grew a clinically significant community-acquired organism, most commonly Salmonella enterica serovar typhi (50.9%), Staphylococcus aureus (19.0%), and Escherichia coli (12.4%). S. aureus bacteremia was common among infants (69.2%), while children 1-5 years had a high frequency of typhoid (44%). Multi-drug-resistant S. Typhi was rare (6%). On multiple logistic regression analysis, typhoid was associated with younger age, longer illness, diarrhea, higher admission temperature, and lower peripheral white blood cell count than non-typhoidal bacteremia. Empirical parenteral ampicillin and gentamicin would have some activity against approximately 88% of clinically significant isolates at a cost of US $1.4/day, an important exception being B. pseudomallei. Bacteremic infants in this setting require an anti-staphylococcal antibiotic. PMID- 17124001 TI - Growth characteristics of ChimeriVax-Den vaccine viruses in Aedes aegypti and Aedes albopictus from Thailand. AB - Four chimeric yellow fever (YF) 17D-dengue (DEN) candidate vaccine viruses (ChimeriVax-DEN; Acambis, Cambridge, MA) were characterized in Aedes aegypti and Ae. albopictus mosquitoes collected from Thailand. The four vaccine viruses contained the relevant prM and E genes of wild-type dengue viruses (DENV; serotypes 1-4) substituted for the equivalent genes in the YF vaccine virus (17D) backbone. Each chimera conferred protection against the homologous DENV serotype; a tetravalent mix of all four chimeras stimulates an immune response against all serotypes. Field-collected mosquitoes from Thailand were fed on blood containing each of the viruses under study and held 21 days after infection. Infection and dissemination rates were based on antigen detection in the body or head tissues, respectively. All four wild-type DENV serotypes infected and disseminated, but the candidate vaccine viruses were highly attenuated in mosquitoes with respect to infection and especially with respect to dissemination. Considering the low level viremias anticipated in humans vaccinated with these viruses, it is predicted that the risks of infection and transmission by mosquitoes in nature is minimal. PMID- 17124002 TI - Geographical distribution of hantaviruses in Thailand and potential human health significance of Thailand virus. AB - Phylogenetic investigations, sequence comparisons, and antigenic cross-reactivity studies confirmed the classification of Thailand virus (THAIV) as a distinct hantavirus species. The examination of sera from 402 rodents trapped in 19 provinces of Thailand revealed that five greater bandicoot rats (Bandicota indica) and one lesser bandicoot rat (B. savilei) from four provinces were focus reduction neutralization test (FRNT) antibody-positive for THAIV. One of 260 patients from Surin province in Thailand (initially suspected of having contracted leptospirosis, but found to be negative) showed symptoms compatible with hemorrhagic fever with renal syndrome (HFRS). The serum of this patient showed high titers of hantavirus-reactive IgM and IgG. FRNT investigations confirmed virus-neutralizing antibodies against THAIV. These observations suggest that THAIV or THAI-like viruses occur throughout Indochina and may represent an additional causative agent of HFRS. PMID- 17124003 TI - Catacamas virus, a hantaviral species naturally associated with Oryzomys couesi (Coues' oryzomys) in Honduras. AB - The primary objective of this study was to extend our knowledge of the geographical distribution, genetic diversity, and natural host associations of the hantaviruses indigenous to North America. Antibody to a hantavirus was found in 5 (20.8%) of 24 Coues' oryzomys (Oryzomys couesi) and none of 41 other rodents captured near the town of Catacamas in eastern Honduras, and a hantavirus was isolated from one of the antibody-positive Coues' oryzomys. Analyses of nucleotide and amino acid sequence data indicated that the viral isolate is a strain of a novel hantaviral species (proposed species name "Catacamas virus") that is phylogenetically most closely related to Bayou virus, a hantaviral species that is principally associated with Oryzomys palustris (marsh oryzomys) in the southeastern United States. Catacamas virus is the first evidence for the occurrence of a hantaviral species in Honduras and the first evidence that a hantaviral species is naturally associated with an Oryzomys species other than O. palustris. PMID- 17124004 TI - In vitro and in vivo studies of ribavirin action on Brazilian Orthobunyavirus. AB - Oropouche, Caraparu, Guama, Guaroa, and Tacaiuma are viruses (genus Orthobunyavirus) that cause human febrile illnesses and encephalitis. The goal of this study was to evaluate the antiviral action of ribavirin on these orthobunyaviruses to achieve a therapeutical agent to treat the diseases caused by these viruses. In vitro results showed that ribavirin (50 microg/mL) had antiviral activity only on the Tacaiuma virus. Addition of guanosine in the culture reversed the antiviral effect of ribavirin on Tacaiuma virus, suggesting that ribavirin inhibited this virus by reducing the intra-cellular guanosine pool. Moreover, ribavirin was not an effective drug in vivo because it was unable to inhibit the death of the mice or virus replication in the brain. The results suggest that ribavirin has no antiviral activity on the Oropouche, Caraparu, Guama, Guaroa, or Tacaiuma viruses; consequently, ribavirin would not be a good therapeutical agent to treat these arboviruses. PMID- 17124005 TI - Short report: HIV infection among commercial sex workers and injecting drug users in the Czech Republic. AB - Since the first HIV case was diagnosed in the Czech Republic in 1985, there is a lack of information regarding the epidemiology of HIV infection in most high-risk groups. To determine the prevalence of, and risk factors for, HIV among female and male commercial sex workers (FCSW and MCSW, respectively) and injecting drug users (IDUs), cross-sectional studies were conducted in the cities of Cheb, Usti nad Labem, Ostrava, and Prague of the Czech Republic. A total of 1,277 subjects were enrolled, which included 585 FCSWs, 230 MCSWs, and 462 IDUs. The HIV prevalences were 0.7% (95% CI: 0.2-1.7%), 0.9% (95% CI: 0.1-3.1%), and 0.2% (95% CI: 0.005-1.2%) among FCSWs, MCSWs, and IDUs, respectively. Although low HIV prevalences were found, ongoing sentinel surveillance studies, which address modifiable behavioral and biologic risk factors among high-risk groups, are necessary to guide strategies to stem the tide of the epidemic in this country. PMID- 17124006 TI - Concise review: stem cells, myocardial regeneration, and methodological artifacts. AB - This review discusses the current controversy about the role that endogenous and exogenous progenitor cells have in cardiac homeostasis and myocardial regeneration following injury. Although great enthusiasm was created by the possibility of reconstituting the damaged heart, the opponents of this new concept of cardiac biology have interpreted most of the findings supporting this possibility as the product of technical artifacts. This article challenges this established, static view of cardiac growth and favors the notion that the mammalian heart has the inherent ability to replace its cardiomyocytes through the activation of a pool of resident primitive cells or the administration of hematopoietic stem cells. PMID- 17124007 TI - Structure and implied functions of truncated B-cell receptor mRNAs in early embryo and adult mesenchymal stem cells: Cdelta replaces Cmu in mu heavy chain deficient mice. AB - Stem cells exhibit a promiscuous gene expression pattern. We show herein that the early embryo and adult MSCs express B-cell receptor component mRNAs. To examine possible bearings of these genes on the expressing cells, we studied immunoglobulin mu chain-deficient mice. Pregnant mu chain-deficient females were found to produce a higher percentage of defective morulae compared with control females. Structure analysis indicated that the mu mRNA species found in embryos and in mesenchyme consist of the constant region of the mu heavy chain that encodes a recombinant 50-kDa protein. In situ hybridization localized the constant mu gene expression to loose mesenchymal tissues within the day-12.5 embryo proper and the yolk sac. In early embryo and in adult mesenchyme from mu deficient mice, delta replaced mu chain, implying a possible requirement of these alternative molecules for embryo development and mesenchymal functions. Indeed, overexpression of the mesenchymal-truncated mu heavy chain in 293T cells resulted in specific subcellular localization and in G(1) growth arrest. The lack of such occurrence following overexpression of a complete, rearranged form of mu chain suggests that the mesenchymal version of this mRNA may possess unique functions. PMID- 17124008 TI - Dynamic compression regulates the expression and synthesis of chondrocyte specific matrix molecules in bone marrow stromal cells. AB - The overall objective of the present study was to investigate the mechanotransduction of bovine bone marrow stromal cells (BMSCs) through the interactions between transforming growth factor beta1 (TGF-beta1), dexamethasone, and dynamic compressive loading. Overall, the addition of TGF-beta1 increased cell viability, extracellular matrix (ECM) gene expression, matrix synthesis, and sulfated glycosaminoglycan content over basal construct medium. The addition of dexamethasone further enhanced extracellular matrix gene expression and protein synthesis. There was little stimulation of ECM gene expression or matrix synthesis in any medium group by mechanical loading introduced on day 8. In contrast, there was significant stimulation of ECM gene expression and matrix synthesis in chondrogenic media by dynamic loading introduced on day 16. The level of stimulation was also dependent on the medium supplements, with the samples treated with basal medium being the least responsive and the samples treated with TGF-beta1 and dexamethasone being the most responsive at day 16. Both collagen I and collagen II gene expressions were more responsive to dynamic loading than aggrecan gene expression. Dynamic compression upregulated Smad2/3 phosphorylation in samples treated with basal and TGF-beta1 media. These findings suggest that interactions between mechanical stimuli and TGF-beta signaling may be an important mechanotransduction pathway for BMSCs, and they indicate that mechanosensitivity may vary during the process of chondrogenesis. PMID- 17124009 TI - Human first-trimester fetal MSC express pluripotency markers and grow faster and have longer telomeres than adult MSC. AB - The biological properties of stem cells are key to the success of cell therapy, for which MSC are promising candidates. Although most therapeutic applications to date have used adult bone marrow MSC, increasing evidence suggests that MSC from neonatal and mid-gestational fetal tissues are more plastic and grow faster. Fetal stem cells have been isolated earlier in development, from first-trimester blood and hemopoietic organs, raising the question of whether they are biologically closer to embryonic stem cells and thus have advantages over adult bone marrow MSC. In this study, we show that human first-trimester fetal blood, liver, and bone marrow MSC but not adult MSC express the pluripotency stem cell markers Oct-4, Nanog, Rex-1, SSEA-3, SSEA-4, Tra-1-60, and Tra-1-81. In addition, fetal MSC, irrespective of source, had longer telomeres (p < .001), had greater telomerase activity (p < .01), and expressed more human telomerase reverse transcriptase (p < .01). Fetal MSC were also more readily expandable and senesced later in culture than their adult counterparts (p < .01). Compared with adult MSC, first-trimester fetal tissues constitute a source of MSC with characteristics that appear advantageous for cell therapy. PMID- 17124010 TI - The TRA-1-60 and TRA-1-81 human pluripotent stem cell markers are expressed on podocalyxin in embryonal carcinoma. AB - We have previously identified the cell adhesion protein podocalyxin expressed in a human pluripotent stem cell, embryonal carcinoma (EC), which is a malignant germ cell. Podocalyxin is a heavily glycosylated membrane protein with amino acid sequence homology to the hematopoietic stem cell marker CD34. Since the initial discovery of podocalyxin in a cancerous stem cell, numerous new studies have identified podocalyxin in many different human cancers and in embryonic stem cells lines (ES) derived from human embryos. Embryonal carcinoma, as do all human pluripotent stem cells, expresses TRA-1-60 and TRA-1-81 antigens, and although their molecular identities are unknown, they are commonly used as markers of undifferentiated pluripotent human stem cells. We report here that purified podocalyxin from embryonal carcinoma has binding activity with the TRA-1-60 and TRA-1-81 antibodies. Embryonal carcinoma cells treated with retinoic acid undergo differentiation and lose the TRA-1-60/TRA-1-81 markers from their plasma membrane surface. We show that podocalyxin is modified in the retinoic acid-treated cells and has an apparent molecular mass of 170 kDa on protein blots as compared with the apparent 200-kDa molecular weight form of podocalyxin expressed in untreated cells. Furthermore, the modified form of podocalyxin no longer reacts with the TRA-1-60/TRA-1-81 antibodies. Thus, embryonal carcinoma expresses two distinct forms of podocalyxin, and the larger version is a molecular carrier of the human stem cell-defining antigens TRA-1-60 and TRA-1-81. PMID- 17124011 TI - Images in clinical medicine. Squamous-cell carcinoma with pericardial metastases. PMID- 17124012 TI - Dangerous deception--hiding the evidence of adverse drug effects. PMID- 17124013 TI - Observational studies of drug safety--aprotinin and the absence of transparency. PMID- 17124014 TI - H5N1 influenza--continuing evolution and spread. PMID- 17124015 TI - Avian influenza A (H5N1) infection in eastern Turkey in 2006. AB - BACKGROUND: An outbreak of highly pathogenic avian influenza A (H5N1) that had previously been detected throughout Asia, with major economic and health repercussions, extended to eastern Turkey in late December 2005 and early January 2006. METHODS: We documented the epidemiologic, clinical, and radiologic features of all cases of confirmed H5N1 virus infection in patients who were admitted to Yuzuncu Yil University Hospital in Van, Turkey, between December 31, 2005, and January 10, 2006. RESULTS: H5N1 virus infection was diagnosed in eight patients. The patients were 5 to 15 years of age, and all eight had a history of close contact with diseased or dead chickens. The mean (+/-SD) time between exposure and the onset of illness was 5.0+/-1.3 days. All the patients had fever, and seven had clinical and radiologic evidence of pneumonia at presentation; four patients died. Results of enzyme-linked immunosorbent assay and rapid influenza tests were negative in all patients, and the diagnosis was made by means of a polymerase-chain-reaction assay. CONCLUSIONS: H5N1, which causes a spectrum of illnesses in humans, including severe and fatal respiratory disease, can be difficult to diagnose. PMID- 17124016 TI - Three Indonesian clusters of H5N1 virus infection in 2005. AB - BACKGROUND: Since 2003, the widespread ongoing epizootic of avian influenza A (H5N1) among poultry and birds has resulted in human H5N1 cases in 10 countries. The first case of H5N1 virus infection in Indonesia was identified in July 2005. METHODS: We investigated three clusters of Indonesian cases with at least two ill persons hospitalized with laboratory evidence of H5N1 virus infection from June through October 2005. Epidemiologic, clinical, and virologic data on these patients were collected and analyzed. RESULTS: Severe disease occurred among all three clusters, including deaths in two clusters. Mild illness in children was documented in two clusters. The median age of the eight patients was 8.5 years (range, 1 to 38). Four patients required mechanical ventilation, and four of the eight patients (50%) died. In each cluster, patients with H5N1 virus infection were members of the same family, and most lived in the same home. In two clusters, the source of H5N1 virus infection in the index patient was not determined. Virus isolates were available for one patient in each of two clusters, and molecular sequence analyses determined that the isolates were clade 2 H5N1 viruses of avian origin. CONCLUSIONS: In 2005 in Indonesia, clusters of human infection with clade 2 H5N1 viruses included mild, severe, and fatal cases among family members. PMID- 17124017 TI - Fetal pulse oximetry and cesarean delivery. AB - BACKGROUND: Knowledge of fetal oxygen saturation, as an adjunct to electronic fetal monitoring, may be associated with a significant change in the rate of cesarean deliveries or the infant's condition at birth. METHODS: We randomly assigned 5341 nulliparous women who were at term and in early labor to either "open" or "masked" fetal pulse oximetry. In the open group, fetal oxygen saturation values were displayed to the clinician. In the masked group, the fetal oxygen sensor was inserted and the values were recorded by computer, but the data were hidden. Labor complicated by a nonreassuring fetal heart rate before randomization was documented for subsequent analysis. RESULTS: There was no significant difference in the overall rates of cesarean delivery between the open and masked groups (26.3% and 27.5%, respectively; P=0.31). The rates of cesarean delivery associated with the separate indications of a nonreassuring fetal heart rate (7.1% and 7.9%, respectively; P=0.30) and dystocia (18.6% and 19.2%, respectively; P=0.59) were similar between the two groups. Similar findings were observed in the subgroup of 2168 women in whom a nonreassuring fetal heart rate was detected before randomization. The condition of the infants at birth did not differ significantly between the two groups. CONCLUSIONS: Knowledge of the fetal oxygen saturation is not associated with a reduction in the rate of cesarean delivery or with improvement in the condition of the newborn. (ClinicalTrials.gov number, NCT00098709 [ClinicalTrials.gov].). PMID- 17124018 TI - Bivalirudin for patients with acute coronary syndromes. AB - BACKGROUND: Current guidelines for patients with moderate- or high-risk acute coronary syndromes recommend an early invasive approach with concomitant antithrombotic therapy, including aspirin, clopidogrel, unfractionated or low molecular-weight heparin, and glycoprotein IIb/IIIa inhibitors. We evaluated the role of thrombin-specific anticoagulation with bivalirudin in such patients. METHODS: We assigned 13,819 patients with acute coronary syndromes to one of three antithrombotic regimens: unfractionated heparin or enoxaparin plus a glycoprotein IIb/IIIa inhibitor, bivalirudin plus a glycoprotein IIb/IIIa inhibitor, or bivalirudin alone. The primary end points were a composite ischemia end point (death, myocardial infarction, or unplanned revascularization for ischemia), major bleeding, and the net clinical outcome, defined as the combination of composite ischemia or major bleeding. RESULTS: Bivalirudin plus a glycoprotein IIb/IIIa inhibitor, as compared with heparin plus a glycoprotein IIb/IIIa inhibitor, was associated with noninferior 30-day rates of the composite ischemia end point (7.7% and 7.3%, respectively), major bleeding (5.3% and 5.7%), and the net clinical outcome end point (11.8% and 11.7%). Bivalirudin alone, as compared with heparin plus a glycoprotein IIb/IIIa inhibitor, was associated with a noninferior rate of the composite ischemia end point (7.8% and 7.3%, respectively; P=0.32; relative risk, 1.08; 95% confidence interval [CI], 0.93 to 1.24) and significantly reduced rates of major bleeding (3.0% vs. 5.7%; P<0.001; relative risk, 0.53; 95% CI, 0.43 to 0.65) and the net clinical outcome end point (10.1% vs. 11.7%; P=0.02; relative risk, 0.86; 95% CI, 0.77 to 0.97). CONCLUSIONS: In patients with moderate- or high-risk acute coronary syndromes who were undergoing invasive treatment with glycoprotein IIb/IIIa inhibitors, bivalirudin was associated with rates of ischemia and bleeding that were similar to those with heparin. Bivalirudin alone was associated with similar rates of ischemia and significantly lower rates of bleeding. (ClinicalTrials.gov number, NCT00093158 [ClinicalTrials.gov].). PMID- 17124019 TI - Educational strategies to promote clinical diagnostic reasoning. PMID- 17124020 TI - The asthma epidemic. PMID- 17124021 TI - Images in clinical medicine. Asymptomatic coronary-artery spasm. PMID- 17124022 TI - Case records of the Massachusetts General Hospital. Case 36-2006. A 35-year-old pregnant woman with new hypertension. PMID- 17124023 TI - Obstetricians still await a deus ex machina. PMID- 17124024 TI - Accounting for ACUITY. PMID- 17124025 TI - Building on experience--the development of clinical reasoning. PMID- 17124026 TI - Research replication. PMID- 17124027 TI - Stress, aging, and neurodegenerative disease. PMID- 17124028 TI - Deep-brain stimulation for Parkinson's disease. PMID- 17124029 TI - Termination of resuscitation in out-of-hospital cardiac arrest. PMID- 17124030 TI - Telithromycin and acute liver failure. PMID- 17124031 TI - Judging the safety of aprotinin. PMID- 17124032 TI - Intracellular protein degradation: from a vague idea thru the lysosome and the ubiquitin-proteasome system and onto human diseases and drug targeting. AB - Between the 1950s and 1980s, scientists were focusing mostly on how the genetic code is transcribed to RNA and translated to proteins, but how proteins are degraded has remained a neglected research area. With the discovery of the lysosome by Christian de Duve it was assumed that cellular proteins are degraded within this organelle. Yet, several independent lines of experimental evidence strongly suggested that intracellular proteolysis is largely non-lysosomal, but the mechanisms involved remained obscure. The discovery of the ubiquitin proteasome system resolved the enigma. We now recognize that degradation of intracellular proteins is involved in regulation of a broad array of cellular processes, such as cell cycle and division, regulation of transcription factors, and assurance of the cellular quality control. Not surprisingly, aberrations in the system have been implicated in the pathogenesis of human disease, such as malignancies and neurodegenerative disorders, which led subsequently to an increasing effort to develop mechanism-based drugs. PMID- 17124033 TI - Immune hemolytic anemia--selected topics. AB - Autoimmune hemolytic anemia (AIHA) is most often idiopathic. However, in recent years, AIHA has been noted with increased incidence in patients receiving purine nucleoside analogues for hematologic malignancies; it has also been described as a complication of blood transfusion in patients who have also had alloimmunization. As the technology of hematopoietic stem cell transplantation has become more widespread, immune hemolysis in the recipients of ABO-mismatched products has become better recognized. The syndrome is caused by passenger lymphocytes transferred from the donor, and although transient, can be quite severe. A similar syndrome has been observed in recipients of solid organ transplants when there is ABO-incompatibility between donor and recipient. Venous thromboembolism is a little-recognized, though likely common, complication of autoimmune hemolytic anemia (AIHA), and may in some instances be related to coexistent antiphospholipid antibodies. While AIHA is a well-documented complication of malignant lymphoproliferative disorders, lymphoproliferative disorders may also paradoxically appear as a consequence of AIHA. A number of newer options are available for treatment of AIHA in patients refractory to corticosteroids and splenectomy. Newer immunosuppressives such as mycophenolate may have a role in such cases. Considerable experience has been accumulating in the last few years with monoclonal antibody therapy, specifically rituximab, in difficult AIHA cases; it appears to be a safe and effective option. PMID- 17124034 TI - Cold hemolytic syndrome. AB - In most cases, immune-mediated hemolysis occurs extravascularly and is associated with IgG antibodies on the surface of red cells. Rare syndromes include IgG antibodies that cause direct intravascular hemolysis, such as paroxysmal cold hemoglobinuria. Also rare are extravascular hemolytic syndromes caused by IgM polyclonal or monoclonal antibodies that demonstrate red cell agglutination at 3 degrees C, so-called cold antibodies. Because cold agglutinin disease has a high association with several lymphoproliferative disorders and IgM monoclonal gammopathies, its management differs significantly from that associated with warm autoimmune hemolytic anemia. This case-based presentation is designed to guide the reader to the diagnosis and to the initiation of prompt, effective therapy. PMID- 17124035 TI - New insights into paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is an uncommon intravascular hemolytic anemia that results from the clonal expansion of hematopoietic stem cells harboring somatic mutations in an X-linked gene, termed PIG-A. PIG-A mutations block glycosylphosphatidylinositol (GPI) anchor biosynthesis, resulting in a deficiency or absence of all GPI-anchored proteins on the cell surface. CD55 and CD59 are GPI-anchored complement regulatory proteins. Their absence on PNH red cells is responsible for the complement-mediated intravascular hemolysis. Intravascular hemolysis leads to release of free hemoglobin, which contributes to many of the clinical manifestations of PNH including fatigue, pain, esophageal spasm, erectile dysfunction and possibly thrombosis. Interestingly, rare PIG-A mutations can be found in virtually all healthy control subjects, leading to speculation that PIG-A mutations in hematopoietic stem cells are common benign events. However, negative selection of PIG-A mutant colony-forming cells with proaerolysin, a toxin that targets GPI-anchored proteins, reveals that most of these mutations are not derived from stem cells. Recently, a humanized monoclonal antibody directed against the terminal complement protein C5 has been shown to reduce hemolysis and greatly improve symptoms and quality of life for PNH patients. PMID- 17124036 TI - Hepcidin and its role in regulating systemic iron metabolism. AB - Maintenance of stable extracellular iron concentrations requires the coordinate regulation of iron transport into plasma from dietary sources in the duodenum, from recycled senescent red cells in macrophages and from storage in hepatocytes. Moreover, during fetal development, the iron requirements of the fetus must be matched by the transport of maternal iron across the placenta. Hepcidin is a 25 amino acid disulfide-rich peptide synthesized in the liver that acts as a systemic iron-regulatory hormone by regulating iron transport from iron-exporting tissues into plasma. Hepcidin inhibits the cellular efflux of iron by binding to, and inducing the degradation of, ferroportin, the sole iron exporter in iron transporting cells. In turn, hepcidin synthesis is increased by iron loading and decreased by anemia and hypoxia. Additionally, hepcidin synthesis is greatly increased during inflammation, trapping iron in macrophages, decreasing plasma iron concentrations and causing iron-restricted erythropoiesis characteristic of anemia of inflammation (anemia of chronic disease). Recent studies indicate that hepcidin deficiency underlies most known forms of hereditary hemochromatosis. This implies that, with the exception of very rare mutations that affect the hepcidin gene itself or modify ferroportin to make it less responsive to hepcidin, hemochromatosis genes encode molecules that regulate hepcidin synthesis. The central involvement of hepcidin in iron regulation and its pathologies should make the eventual hepcidin assay useful for the diagnosis of iron disorders and the monitoring of their treatments. The development of hepcidin agonists and antagonists may provide useful therapeutics for the treatment of iron disorders. PMID- 17124037 TI - Current approaches to the management of hemochromatosis. AB - The term hemochromatosis encompasses at least four types of genetic iron overload conditions, most of them recently distinguished from one another as a result of the identification of a series of genes related to iron metabolism. At least three of these entities (HFE hemochromatosis, juvenile hemochromatosis and transferrin receptor 2 hemochromatosis) involve systemic hepcidin deficiency as a key pathogenetic factor. Major advances in the management of hemochromatosis influence the diagnostic approach to the disease, with the development of an overall non invasive strategy, mainly based on clinical, biological (iron parameters and genetic testing), and imaging (especially magnetic resonance imaging) data. Therapeutic management remains, on the curative side, dominated by phlebotomy (venesection), practical aspects of which have been recently revisited by the Guidelines Department of the French "Haute Autorite de Sante." However, innovative treatment approaches, based on the improved pathophysiological understanding of these diseases and the progress in iron chelation therapy, are emerging. Preventive therapy, focused on family screening, remains a key part of the management of hemochromatosis. PMID- 17124038 TI - New advances in iron chelation therapy. AB - The emergence of new chelators is likely to have a major impact on the treatment of thalassemia major, sickle cell disease and other hematologic disorders for which regular red cell transfusions are required either to correct severe anemia or to prevent major complications of the underlying disease. In comparison with deferoxamine, which requires prolonged parenteral infusion to achieve negative iron balance, orally active chelators offer the promise of easier administration and better compliance. Moreover, the availability of more than one iron chelator opens up the possibility of combination therapy to target iron in specific organs and to enhance overall iron excretion. Long-term studies, employing new technologies to measure tissue iron, are required to determine whether the new chelators will be as safe and effective as deferoxamine in maintaining control of iron stores and preventing iron-induced cardiac disease, the major complication of transfusional iron overload. PMID- 17124039 TI - Red cell exchange in sickle cell disease. AB - Red cell exchange transfusions remain an effective but possibly underutilized therapy in the acute and chronic treatment of sickle cell disease. In sickle cell disease, increased blood viscosity can cause complications when the hemoglobin exceeds 10 g/dL even if this is due to simple transfusion. Red cell exchange can provide needed oxygen carrying capacity while reducing the overall viscosity of blood. Acute red cell exchange is useful in acute infarctive stroke, in acute chest and the multi-organ failure syndromes, the right upper quadrant syndrome, and possibly priapism. Neither simple or exchange transfusions are likely to hasten resolution of an acute pain episode. PMID- 17124040 TI - Prevention and management of stroke in sickle cell anemia. AB - As the overall health of patients with sickle cell anemia (SS) improves and diagnostic techniques become more sensitive, physicians are seeing patients with an increasingly wide range of subtle and not-so-subtle brain injury. The major breakthrough in the field of sickle-related brain injury has been the unprecedented success of transcranial Doppler ultrasonography (TCD) to identify asymptomatic patients at high risk of stroke, coupled with chronic transfusion therapy to prevent it. The evidence for TCD screening and preventive treatment is strong and compelling, but there are still important unanswered questions regarding the implications of "silent infarcts" found in the magnetic resonance images (MRIs) of asymptomatic individuals, and the growing awareness of the burden of neuropsychiatric dysfunction in otherwise apparently healthy individuals. PMID- 17124041 TI - Induction of fetal hemoglobin in the treatment of sickle cell disease. AB - Reactivation of fetal hemoglobin (HbF) expression is an important therapeutic option in patients with hemoglobin disorders. In sickle cell disease (SCD), an increase in HbF inhibits the polymerization of sickle hemoglobin and the resulting pathophysiology. Hydroxyurea, an inducer of HbF, has already been approved for the treatment of patients with moderate and/or severe SCD. Recent clinical trials with other pharmacological inducers of HbF, such as butyrate and decitabine, have shown considerable promise. In this chapter, we highlight the important clinical trials with pharmacological inducers of HbF, discuss their mechanisms of action and speculate about the future of this therapeutic approach in the treatment of patients with SCD. PMID- 17124042 TI - Inherited bone marrow failure syndromes: molecular features. AB - Recent advances resulting from the identification of the genes responsible for four inherited marrow failure syndromes, Fanconi anemia, dyskeratosis congenita, Diamond-Blackfan anemia, and Shwachman-Diamond syndrome, are reviewed. The interpretation of genetic testing should be guided by an understanding of the limitations of such testing for each disorder. The possibility of an inherited basis for marrow failure must be considered for adults as well as children with aplastic anemia. Shared molecular themes are emerging from functional studies of the genes underlying the different inherited disorders. Genomic instability may result from impaired DNA repair in Fanconi anemia or telomere dysregulation in dyskeratosis congenita. Mutations affecting ribosome assembly or function are associated with Diamond-Blackfan anemia, dyskeratosis congenita, and Shwachman Diamond syndrome. These findings raise new questions about the molecular mechanisms regulating hematopoiesis and leukemogenesis. Clinical implications arising from these molecular studies are explored. PMID- 17124043 TI - Pathophysiologic mechanisms in acquired aplastic anemia. AB - Aplastic anemia, an unusual hematologic disease, is the paradigm of the human bone marrow failure syndromes. Absence of hematopoietic cells has been recognized from the characteristic morphology for a century; an immune pathophysiology has been inferred from improvement in blood counts with immunosuppressive therapy in the majority of patients. Molecular mechanisms underlying both T cell effector cells and the target marrow stem and progenitor cells are now being identified. Activated type 1 cytotoxic T cells and type 1 cytokines have been implicated in cell culture experiments; clues to the molecular basis of the aberrant immune response include cytokine gene polymorphisms and abnormalities in the regulatory pathways for gamma-interferon. For stem cell depletion, mutations in genes of the telomere repair complex are present in some patients with apparently acquired aplastic anemia. Telomerase deficiency is associated with short telomeres and a quantitative reduction in marrow progenitors and likely also a qualitative deficiency in the repair capacity of hematopoietic tissue. PMID- 17124044 TI - Making therapeutic decisions in adults with aplastic anemia. AB - The management of adults presenting with aplastic anemia (AA) requires careful exclusion of other causes of bone marrow failure. Late-onset inherited forms of AA may present in adulthood with subclinical disease. Recent long-term studies of HLA identical sibling donor BMT show excellent survival for patients under the age of 40 years, but chronic graft-versus-host disease (GVHD) is still a major problem, impacting on quality of life. Recent improvements in outcome after matched unrelated donor BMT may reflect better donor matching and use of reduced intensity conditioning regimens. For patients treated with immunosuppressive therapy (IST), antithymocyte globulin (ATG) and cyclosporin (CSA) remain the standard regimen with excellent overall survival but less impressive failure-free survival due to nonresponse, relapse and later clonal disorders. The benefit of adding granulocyte colony-stimulating factor (G-CSF) to ATG and CSA is unclear and being assessed in a further prospective European study. Patients who are refractory to conventional IST and currently ineligible for BMT represent difficult management problems. For these patients, new approaches to transplantation are being evaluated, such as fludarabine-based conditioning regimens and the potential use of double umbilical cord blood transplants, but there is a need for new immunosuppressive agents. Improved supportive care is likely to be a major factor in improved outcome of all AA patients whether treated with IST or BMT. Robust predictive factors for response to IST are needed to help in decision making at diagnosis and to help justify exploring novel approaches to therapy. PMID- 17124045 TI - Thrombosis in infants and children. AB - During the last decade much progress has been made toward better understanding of the underlying reasons causing thromboembolism in children. A considerable number of acquired and hereditary thrombotic risk factors have been identified which may also have an impact on therapeutic decisions and prognosis concerning outcome and the risk of a second event. However, indications for therapeutic interventions, such as thrombolysis and prophylactic anticoagulation with respect to the different clinical conditions and their combination with other risk factors, are not yet well defined. The following article describes the causes, clinical presentation and management of thrombosis in neonates, infants and older children, focusing on the clinically most relevant conditions. PMID- 17124046 TI - Immune thrombocytopenic purpura of childhood. AB - Immune mediated thrombocytopenia (ITP) is a common manifestation of autoimmune disease in children. Although patients often present with bruises, petechiae, and some mucosal bleeding, the incidence of life-threatening hemorrhage is rare (0.2 0.9%) but can be fatal when presenting in vital organs. A wide range of therapeutic regimens are currently in use, including observation alone, as the majority of children recover within 4-6 months regardless of treatment. A growing understanding of the pathophysiology of acute ITP in children has not impacted the controversy surrounding treatment, but has clarified the mechanism of action of the most frequently used agents in chronic ITP. Newer monoclonal antibodies such as Rituxan have proved very useful in chronic or refractory ITP and studies are ongoing to determine the best regimens using this form of immune modulation. Splenectomy and newer agents to boost platelet production are also under study in chronic ITP. Neonates may also have a form of immune thrombocytopenia with extensive bruising and thrombocytopenia called neonatal alloimmune thrombocytopenic purpura (NATP). Rather than autoantibodies, the platelet destruction is secondary to transplacental maternal IgG alloantibodies. During pregnancy mothers may become sensitized to platelet membrane antigens present on fetal platelets. These antibodies may result in serious bleeding, including intracranial hemorrhage in the perinatal period. Once identified, these mothers may require treatment during future pregnancies to minimize serious bleeding in the fetus and neonate. Treatment in utero and immediately following delivery is focused on restoring neonatal platelets to a safe level and preventing life threatening bleeding. PMID- 17124047 TI - Disorders of neutrophil number and function. AB - This review of disorders of neutrophil number and function will discuss important research advances in the field and then provide a clinical diagnostic approach. The focus will be on two recent clinical developments in the field of phagocyte disorders. First, an important natural history study from the Severe Chronic Neutropenia International Registry has recently quantitated the incidence and risk factors for death from sepsis and for progression to myelodysplastic syndrome and acute myeloid leukemia in a large cohort of severe chronic neutropenia patients, many of whom were followed 10 or more years on treatment with granulocyte colony-stimulating factor. Second, in the past year, a multinational group has announced successful gene therapy of two adults with chronic granulomatous disease, the most common disorder of neutrophil function. However, monitoring of retroviral insertion sites revealed expansion of the multiclonal population of gene-modified cells, raising concerns about eventual leukemogenesis. The review also provides a pragmatic approach to the evaluation of a patient with a suspected disorder of neutrophil number or function. PMID- 17124048 TI - Pharmacogenetics, pharmacogenomics and personalized medicine: are we there yet? AB - The genetic basis of a differential response to drugs has been understood for a limited number of agents for over 30 years. This knowledge has generated hope that the individual basis for response to a wide range of drugs would be quickly known, and individualized drug selection and dosing would be possible for many or all disorders. Understanding the variable response to drugs seems particularly pressing in the field of oncology, in which the stakes are high (failure to cure cancer usually leads to death), drugs commonly have a narrow therapeutic index, and toxicities can be severe (a significant frequency of toxic death is a feature of most acute myeloid leukemia protocols, for example). However, in common with many new technologies, the generalizability and clinical application of pharmacogenetics has proved more challenging than expected. Difficulties include, in many examples, a modest clinical effect relative to genotype, therapy specific, not broad, applicability and the very major challenge of unraveling the complexity of gene-gene interactions. In addition, ethical and economic challenges to the application of pharmacogenetics have moved to the fore in recent years, particularly in the context of racial differences in outcome of therapy. Genomic, rather than candidate gene approaches to identification of relevant loci are increasingly being explored, and significant progress is being made. However, greater understanding of the complexities of multiple gene modifiers of outcome, and the statistical challenge of understanding such data, will be needed before individualized therapy can be applied on a routine basis. PMID- 17124049 TI - Tumor-specific genetic lesions and their influence on therapy in pediatric acute lymphoblastic leukemia. AB - Pharmacogenomics has traditionally focused on the identification of inherited genetic differences that influence a patient's response to a specific therapeutic agent. These differences can range from inherited variability in the genes that affect drug absorption, distribution, intracellular transport, metabolism, and elimination, to variability in the genes that encode either the target of the drug or components of the pathway affected by the drug. The main goal of pharmacogenomics is to improve our understanding of how these variations, either individually or collectively, influence the therapeutic response. The genetic differences inherent within cancer cells constitute the other major variable in a patient's ultimate response to therapy. In this review, we provide an overview of high-throughput genomic methods that can be used to identify genetic lesions within cancer cells. These efforts will ultimately allow the identification of the full complement of genetic lesions that underlie the establishment and maintenance of the leukemic clone. The identification of these lesions should provide the bases for defining the molecular "Achilles heels" against which new targeted therapies can be developed. PMID- 17124050 TI - Deeper insights into hematological oncology disorders via single-cell phospho signaling analysis. AB - An understanding of hematological cancer cell signaling processes poses one of the most complex and intractable problems in modern biomedical inquiry. While we understand some of the fundamental players that contribute to oncogenic processes, significant effort is focused upon determining how these individual players relay information to each other to create the composite functions of a cancer cell. Efforts designed to understand these processes at the single cell level will undoubtedly allow for understanding of the heterogeneity of hematological tumors as well as, simultaneously, the function of the 'responding' immune system. I will relate some of the insights our laboratory has developed over the last several years applying single-cell phospho-flow cytometry to the study of signaling in primary patient material and murine models. While it is clear that this analysis now allows us to accomplish phospho-signaling biochemistry at the single cell level with primary cell material, we are only beginning to develop some of the bioinformatics tools to appropriately display the vast amount of information collected by such approaches. These approaches, however, have already allowed us to develop approaches that prognosticate patient outcomes based on signaling status, prior to any treatment, as well as subgroup patient subtypes according to signaling states. The modest efforts to date presage a time where it should be possible to provide far more tailored therapies specific to the varied diseases represented by the hematological malignancies. PMID- 17124051 TI - Myths and lessons from the adult/pediatric interface in acute lymphoblastic leukemia. AB - The development of effective therapy for children with acute lymphoblastic leukemia (ALL) is one of the great successes of clinical oncology, with long-term survival achieved in over 80% of patients. However, cure rates for adults with ALL remain relatively low, with only 40% of patients cured. With an age unrestricted, biology-based approach, we anticipate a better understanding about why these outcome differences exist, and think that by extending successful pediatric clinical programs to include adult patients with ALL, we can directly compare uniformly treated adults and children in terms of response to therapy, toxicity and underlying biology. PMID- 17124052 TI - Treatment of adult acute lymphoblastic leukemia. AB - In the early 1980s, adult acute lymphoblastic leukemia (ALL) was a rarely curable disease with overall survival < 10%. After adapting combinations employed by pediatric groups, the outcome improved to 30-40%. A period of stagnation followed with improvement only in distinct subgroups. In the past 5 years, however, striking new developments have been noticeable. Progress has been made in molecular diagnostics of ALL. Improvements to standard therapy including stem cell transplantation (SCT) have occurred and a variety of new drugs for ALL are under evaluation. Rapid diagnosis and classification of ALL is increasingly important to identify prognostic factors and molecular genetic subsets that will be the focus of "targeted" therapies as we enter the era of subset specific treatment. In the following review we will discuss treatment of adult ALL (excluding elderly patients,(1) adolescents(2) and patients with Ph/BCR-ABL positive ALL(3)). PMID- 17124053 TI - Central nervous system disease in acute lymphoblastic leukemia: prophylaxis and treatment. AB - Improved treatment for acute lymphoblastic leukemia (ALL) has virtually eliminated testicular relapse. However, the control of central nervous system (CNS) leukemia remains a therapeutic challenge in childhood ALL, partly because of the late complications arising from cranial irradiation. In most current pediatric protocols, cranial irradiation (12 to 18 Gy) is given to 5% to 25% of patients--those with T-cell ALL, overt CNS disease (CNS3 status) or high-risk cytogenetics. CNS control is a less urgent concern in adults with ALL, in whom systemic relapse remains the major problem. With current approaches, approximately 2% to 10% of patients can be expected to develop CNS relapse. Children with B-cell precursor ALL who have a late CNS relapse (after an initial remission of 18 months or more) and did not receive cranial irradiation have an excellent outcome after retrieval therapy, with a 5-year event-free survival (EFS) rate approaching that in newly diagnosed patients. Innovative treatment options are needed for children who develop CNS relapses after a short initial remission or after receiving cranial irradiation, and in any adults with CNS leukemia at diagnosis or relapse. PMID- 17124054 TI - Treatment of acute promyelocytic leukemia. AB - Cure of acute promyelocytic leukemia (APL) is now a possibility for most patients through the use of state-of-the-art treatments, which include simultaneous administration of all-trans retinoic acid (ATRA) and anthracycline-based chemotherapy for induction and consolidation, as well as ATRA-based maintenance. Risk-adapted strategies to modulate treatment intensity may be an effective approach to minimize therapy-related morbidity and mortality while maintaining the potential of cure. In this context, there is no role for hematopoietic stem cell transplantation (HSCT) in front-line therapy, except for the small fraction of patients with persistent minimal residual disease at the end of consolidation. However, HSCT plays an important role for patients in second complete remission. In contrast, an increasing role of arsenic trioxide (ATO) is emerging. Given the high antileukemic efficacy observed with ATO in patients relapsing after ATRA containing regimens, this agent is currently regarded as the best treatment option in this setting. However, until a randomized comparison between the standard therapy and ATO-based regimens in front-line therapy is available, this latter approach should only be recommended for unfit patients for whom chemotherapy is contraindicated. In addition to reviewing current consensus and controversial issues on antileukemic strategies, this review addresses other aspects that can be crucial for the outcome of individual patients. These aspects include supportive care, recognition and treatment of life-threatening complications, evaluation of response, and, finally, management of the disease in special conditions such as older patients, children and pregnant women. PMID- 17124055 TI - The biology of acute promyelocytic leukemia and its impact on diagnosis and treatment. AB - Several genetic and phenotypic characteristics of acute promyelocytic leukemia (APL) blasts provide relevant targets and the rationale for tailored treatment. These include the PML/RARalpha fusion and the transcription co-repressor complex recruited at the promoter of target genes by the hybrid protein, the intense and homogeneous expression of the CD33 antigen, absence of multidrug resistance related phenotype, and a frequently mutated and constitutively activated FLT3 receptor. Such genotypic and phenotypic features are targeted by agents currently in use in front-line therapy or at relapse (i.e., retinoids, arsenic trioxide, anthracyclines and anti-CD33 monoclonal antibodies), and by novel agents that may find a place in future treatments such as histone deacetylase and FLT3 inhibitors. The unique PML/RARalpha aberration serves as a molecular marker for rapid diagnosis and prediction of response to ATRA-and ATO-containing therapies. Methods for prompt and low-cost detection of this genetic abnormality, such as the analysis of PML nuclear staining, are extremely useful in clinical practice and could be adopted in countries with limited resources as a surrogate for rapid genetic diagnosis. Finally, PML/RARalpha monitoring through sensitive RT-PCR can be regarded as an integrating part of the overall treatment strategy in this disease, whereby the treatment type and intensity are modulated in patients at different risk of relapse according to RT-PCR status during follow-up. Because recent clinical studies suggest that most APL patients receiving intensive chemotherapy may be over-treated, longitudinal and stringent RT-PCR monitoring is becoming increasingly important to test the extent to which chemotherapy can be minimized in those presenting with low-risk disease. PMID- 17124056 TI - Management of APL in developing countries: epidemiology, challenges and opportunities for international collaboration. AB - Acute promyelocytic leukemia (APL), a relatively rare hematologic malignancy, is highly curable with current treatment strategies. However, these strategies may be unavailable in countries with limited resources. A review of records in several Latin American countries revealed that approximately 30% of deaths among children and adults with APL were caused by early complications associated with the disease or its treatment. Further, APL accounts for 20% to 25% of cases of AML in these countries, consistent with the previous observation of increased incidence of APL in Latin Americans. The lack of population-based registries in developing countries has made it difficult to determine the real incidence of APL. Moreover, APL appears to have other unique epidemiologic characteristics, including association of primary APL with an increased body mass index at diagnosis and association of secondary APL with breast cancer. To facilitate the development of local capacity and implement effective treatment of APL in developing countries, the International Committee of the American Society of Hematology has assembled a working group to formulate treatment guidelines based on evidence from clinical trials results in the developed world but adapted to local resources. It is hoped that uniform treatment, careful documentation of specific outcome data, and ongoing monitoring of treatment efficacy and toxicity will improve the cure rate and provide biologic and epidemiologic information about APL in developing countries. This initial demonstration project may be joined by other countries, providing a framework for additional clinical investigation in this highly curable form of leukemia. PMID- 17124057 TI - Chromosome aberrations, gene mutations and expression changes, and prognosis in adult acute myeloid leukemia. AB - Pretreatment clinical features and prognosis of patients with acute myeloid leukemia (AML) are strongly influenced by acquired genetic alterations in leukemic cells, which include microscopically detectable chromosome aberrations and, increasingly, submicroscopic gene mutations and changes in gene expression. Cytogenetic findings separate AML patients into three broad prognostic categories: favorable, intermediate and adverse. The cytogenetic-risk classifications differ somewhat for younger adult patients and those aged 60 years or older. In many instances, patients with specific cytogenetic findings, e.g., those with a normal karyotype or those with either t(8;21)(q22;q22) or inv(16)(p13q22)/t(16;16)(p13;q22) [collectively referred to as core-binding factor (CBF) AML] can be further subdivided into prognostic categories based on the presence or absence of particular gene mutations or changes in gene expression. Importantly, many of these molecular genetic alterations constitute potential targets for risk-adapted therapies. In this article, we briefly review major cytogenetic prognostic categories and discuss molecular genetic findings of prognostic significance in two of the largest cytogenetic groups of patients with AML, namely AML with a normal karyotype and CBF AML. PMID- 17124058 TI - FLT3 mutations: biology and treatment. AB - FLT3 is a receptor tyrosine kinase with important roles in hematopoietic stem/progenitor cell survival and proliferation. It is mutated in about 1/3 of acute myeloid leukemia (AML) patients, either by internal tandem duplications (ITD) of the juxtamembrane domain or by point mutations usually involving the kinase domain (KD). Both types of mutation constitutively activate FLT3. Many studies have shown that AML patients with FLT3/ITD mutations have poor cure rates due to relapse. This has led to the development of a number of small molecule tyrosine kinase inhibitors (TKI) with activity against FLT3. Many of these are still in preclinical development, but several have entered clinical phase I and II trials as monotherapy in patients with relapsed AML. Patients with FLT3 mutations in these trials have shown clinical responses, most often a clearing of peripheral blasts, but rarely major reductions of bone marrow blasts. Several studies have shown that FLT3 was successfully inhibited in most patients. However, complete remissions have rarely been achieved in these trials. The difference in responses of chronic myeloid leukemia (CML) patients to BCR-ABL inhibitors compared to FLT3 mutant AML patients to FLT3 inhibitors may be reflective of treating a single gene disease in CML versus multiply altered gene disease in AML. This has led to clinical testing of FLT3 TKI in combination with conventional chemotherapy, with trial designs based on preclinical testing showing synergistic effects between these agents in inducing cytotoxic responses. Several combination trials are ongoing or planned in both relapsed and newly diagnosed FLT3-mutant AML patients. PMID- 17124059 TI - Controversies in treatment of AML: case-based discussion. AB - Treatment of acute myeloid leukemia (AML) in older adults remains a tremendous challenge. Standard approaches to treatment have resulted in progression-free survival in only a small minority of patients with AML over the age of 60. Elucidation of the molecular genetic events that contribute to the pathogenesis of AML in older patients are providing insights into mechanisms of resistance. This knowledge is also providing new opportunities to begin to refine and direct novel therapies for these heterogeneous diseases. In this case-based review, prognostic factors for treatment outcome in older adults will be covered along with discussion of a risk-based approach to potential therapeutic options, ranging from palliative care to novel therapies and reduced-intensity allogenic transplant. PMID- 17124060 TI - Treatment of the 5q- syndrome. AB - Defined by isolated del 5q and no excess of marrow blasts, the "5q- syndrome" is a specific type of myelodysplastic syndrome (MDS) with particular characteristics, including severe anemia, frequent thrombocytosis, typical dysmegakaryopoiesis and favorable outcome. Its pathogenesis remains uncertain, in particular the role of inactivation of gene(s) situated in 5q. It should be differentiated from other MDS with del 5q having an excess of marrow blasts and/or additional cytogenetic abnormalities, which carry a poor prognosis. Until the advent of lenalidomide, repeated RBC transfusions were generally the only treatment of the 5q- syndrome, which was resistant to other therapeutic approaches. Lenalidomide can lead to RBC transfusion independence in at least two thirds of cases of the 5q- syndrome, two thirds of those responses persisting after 2 years of treatment. Importantly, not only reversal of anemia but also frequent complete pathological and cytogenetic responses are obtained. Grade 3 or 4 neutropenia and thrombocytopenia, especially during the first 6 to 8 weeks of treatment, are the major side effect of lenalidomide, justifying close monitoring of blood counts and regular patient visits. Preliminary results suggest that lenalidomide is also very active in MDS with del 5q other than the 5q-syndrome. Although its mechanism of action remains uncertain, lenalidomide appears to target specifically the del 5q clone. By doing this, lenalidomide may have an effect on disease course and survival, which is currently being assessed in clinical trials. PMID- 17124061 TI - Hematopathological concepts and controversies in the diagnosis and classification of myelodysplastic syndromes. AB - Although the diagnosis and classification of most cases of the myelodysplastic syndromes (MDS) is usually accomplished without difficulty, a minority of cases may pose diagnostic problems. In many cases the diagnostic dilemma can be solved by adhering to basic guidelines recommended for evaluation of patients suspected of having MDS, and in particular to the quality of the blood and bone marrow specimens submitted for morphologic, immunophenotypic and genetic studies. In other cases, such as patients who have hypocellular MDS or MDS with fibrosis, the criteria for making a diagnosis may be difficult if not impossible to apply, and in still others the diagnostic uncertainty is because the minimal criteria necessary to establish the diagnosis of MDS are not always clearly stated. In this review, some of these diagnostic problems are addressed and some general guidelines for resolving them are suggested. In addition, data are presented that illustrate that the WHO classification offers a valuable tool in the diagnosis and classification of MDS. PMID- 17124062 TI - Clinical issues in the management of patients with myelodysplasia. AB - The management of patients with myelodysplasia (MDS) can be quite complex and varies according to both the clinical manifestations in individual patients as well as complicating medical conditions. Allogeneic stem cell transplantation is the only curative treatment, but because of the older age of the patient population must be applied selectively, particularly in those with lower risk MDS as well as in patients whose clinical course is more frankly "preleukemic." Issues pertinent to the use of 5-azacytidine, decitabine and lenalidomide in patients with both higher and lower International Prognostic Staging System (IPSS) stage disease are discussed. PMID- 17124063 TI - ABL kinase inhibitor therapy for CML: baseline assessments and response monitoring. AB - For chronic phase chronic myeloid leukemia (CML) patients treated with imatinib, the essential pre-therapy assessments include bone marrow morphology and cytogenetics as well as a baseline real-time quantitative PCR (RQ-PCR) for BCR ABL. Baseline assessments of clinical relevance include Sokal and Hasford prognostic scores. Several other baseline assays are likely to be predictive of response, including IC50(imatinib), organic cation transporter-1 (OCT-1) mRNA level, and gene expression profiles, but further confirmation is required. RQ-PCR assays of blood at least every 3 months once patients have commenced imatinib is recommended. This will facilitate early identification of suboptimal responders who may benefit from higher doses of imatinib or alternative therapy, and identify at an early stage patients with acquired resistance. Management of the latter group can be further guided by the findings from cytogenetics and BCR-ABL kinase domain mutation screening. Bone marrow cytogenetics is indicated at least every 6 months until the patient achieves major molecular response. RQ-PCR is only clinically useful if it is conducted under a rigorous quality control regimen so that fluctuations in the BCR-ABL level can be confidently attributed to a biological cause rather than assay variation. To further improve the clinical value of RQ-PCR monitoring, expression of results on an international scale is needed. This will facilitate a more uniform and rational approach to management of suboptimal response and loss of response. PMID- 17124064 TI - Defining and managing imatinib resistance. AB - While imatinib is highly effective therapy, with improving prospects over time for sustained remission and potential to severely limit or eliminate disease progression and transformation, a minority of patients either fail or respond suboptimally to imatinib; as well, disease eradication may not be possible with imatinib. Distinct patterns of resistance have evolved with the use of imatinib, and Abl kinase mutations, which alter imatinib binding or favor kinase conformations inaccessible to imatinib, are a common finding associated with clinical resistance. Dasatinib and nilotinib, alternate Abl kinase inhibitors, restore hematologic and cytogenetic remission in the majority of patients with primary failure or acquired resistance in chronic phase disease; in advanced disease and Philadelphia chromosome (Ph)(+) ALL, responses are more limited and relapse is common. Future studies with these agents will focus on further optimizing imatinib response, reduction of minimal residual disease, and prevention of resistance. Still newer inhibitors active against T315I mutant BCR ABL may overcome primary and secondary resistance to dasatinib and nilotinib. PMID- 17124065 TI - Managing the patient with chronic myeloid leukemia through and after allogeneic stem cell transplantation. AB - Although the only curative therapy for chronic myeloid leukemia remains allogeneic stem cell transplantation (allo-SCT), early to mid-term results of imatinib in newly diagnosed patients are sufficiently impressive to have displaced allo-SCT to second- or third-line treatment. Patients now arrive at a decision for transplantation in a variety of disease situations: failing to achieve certain hematological, cytogenetic and molecular milestones by some pre determined timepoint, having lost a previous best response or by progression to advanced phase. The decision, therefore, is not simply whether to transplant or not, but also how to transplant. Evolving transplant technology requires that the individual circumstances of each patient should be considered when recommending the procedure. Attempts to improve the safety of transplant are generally associated with a reduction in long-term disease control and patient monitoring, and management is life-long. The treatment of recurrent disease is no longer straightforward, with the choices being donor lymphocytes or tyrosine kinase inhibitors alone or in combination. This section will review the evidence supporting some of these decisions and highlight current controversies. PMID- 17124066 TI - Role of JAK-STAT signaling in the pathogenesis of myeloproliferative disorders. AB - The identification of JAK2V617F mutations in polycythemia vera (PV), essential thrombocytosis (ET), and myelofibrosis (MF) represents an important advance in our understanding of these myeloproliferative disorders (MPD). Most, if not all, patients with PV and a significant number of patients with ET and MF are JAK2V617F positive, and the mutation likely arises in the hematopoietic stem cell compartment. JAK2V617F is a constitutively active tyrosine kinase that is able to activate JAK-STAT signaling most efficiently when co-expressed with the erythropoietin receptor (EPOR), the thrombopoietin receptor (MPL), or the granulocyte colony-stimulating factor receptor (GCSFR). Data from murine models supports the central role of JAK2V617F in the pathogenesis of MPD, as expression of JAK2V617F in a bone marrow transplantation assay results in polycythemia and myelofibrosis in recipient mice. Activation of JAK-STAT signaling by JAK2V617F in some, but not all MPD patients with ET and MF led to the identification of the constitutively active MPLW515L allele in ET and MF. Small molecule inhibitors of JAK-STAT signaling are currently being developed, which offer potential for molecularly targeted therapy for patients with PV, ET, and MF. Despite these advances, many questions remain regarding the role of a single disease allele in three phenotypically distinct MPD, the potential clinical efficacy of JAK2 inhibitors, and the identity of oncogenic alleles in JAK2V617F/MPLW515-negative MPD. PMID- 17124067 TI - Classification, diagnosis and management of myeloproliferative disorders in the JAK2V617F era. AB - JAK2V617F, a somatic gain-of-function mutation involving the JAK2 tyrosine kinase gene, occurs in nearly all patients with polycythemia vera (PV) but also in a variable proportion of patients with other myeloid disorders; mutational frequency is estimated at approximately 50% in both essential thrombocythemia (ET) and myelofibrosis (MF), up to 20% in certain subcategories of atypical myeloproliferative disorder (atypical MPD), less than 3% in de novo myelodysplastic syndrome (MDS) or acute myeloid leukemia, and 0% in chronic myeloid leukemia (CML). Accordingly, there is now molecular justification for grouping PV, ET, and MF together in a distinct MPD category (i.e., classic, BCR ABL(-) MPD) that is separate from chronic myeloid leukemia (CML), MDS, and atypical MPD. To date, JAK2V617F has not been described in patients with reactive myeloproliferation, lymphoid disorders, or solid tumor. Therefore, the presence of JAK2V617F strongly suggests an underlying MPD and it is therefore reasonable to consider JAK2V617F-based laboratory tests for the evaluation of polycythemia, primary thrombocytosis, unexplained leukocytosis, bone marrow fibrosis, or abdominal vein thrombosis. Current information on disease-specific prognostic relevance of JAK2V617F is inconclusive and confounded by inter-study differences in the performance of mutation screening assays. Regardless, the discovery of JAK2V617F has reinforced the pathogenetic contribution of JAK-STAT signaling in MPD and identifies JAK2 as a valid drug target. PMID- 17124068 TI - Myeloproliferative disease in pregnancy and other management issues. AB - The goal of this review is to assess the current treatment and outcomes of special clinical situations in patients with myeloproliferative disease (MPD) such as pregnancy, major thrombotic and bleeding complications and surgical interventions. However, only a limited literature to support optimal management strategies is available. Many of the proposed strategies are the results of common sense or derive from the extrapolation of data from other studies not specifically designed to solve these problems. Therefore, practical recommendations to guide clinical decisions in these settings still remain largely empirical. PMID- 17124069 TI - The management of patients with limited-stage classical Hodgkin lymphoma. AB - The term limited-stage Hodgkin lymphoma refers to those patients with stage I-II disease and an absence of bulky disease. Among those patients with classical Hodgkin lymphoma, approximately one-third of patients will fall into this category. As long-term disease control can now be anticipated in more than 90% of these patients, management strategies must increasingly address the need to reduce the long-term treatment-related risks. Current treatment options include use of combined modality therapy that includes an abbreviated course of chemotherapy and involved-field radiation or treatment with chemotherapy, currently consisting of ABVD, as a single modality. The choice of treatment between these two options involves specific trade-offs that must balance issues of disease control against long-term risk of late effects. PMID- 17124070 TI - Utility of positron emission tomography (PET) scanning in managing patients with Hodgkin lymphoma. AB - Use of positron emission tomography (PET) or PET/ computed tomography (CT) in Hodgkin lymphoma (HL) continues to expand worldwide. PET is currently widely utilized for response assessment after completion of therapy and, to a lesser extent, for pretreatment staging and assessment of response during therapy (therapy monitoring). In pretreatment staging, PET cannot replace CT or bone marrow biopsy (BMB); however, it can provide complementary information to both CT and BMB, potentially resulting in a modification of disease stage (usually upstaging) in about 15-20% of patients with impact on management in about 5-15%. PET for response assessment at the conclusion of treatment is substantially more accurate than CT because of its ability to distinguish between viable tumor and necrosis or fibrosis in posttherapy residual mass (es) that are present in about two-thirds of patients with HL without any other clinical or biochemical evidence of disease. PET, therefore, provides more accurate response classifications compared with CT-based assessment. The role of PET for therapy monitoring is still evolving but may prove to be the most exciting with potentially high impact on patient management and outcome. PET evaluation during therapy appears to be at least as accurate for predicting patient outcome as evaluation after completion of therapy and its use is clearly justified if the purpose is to provide an early and yet accurate assessment of response with the clear intent of tailoring therapy according to the information provided by the scan. The role of PET scanning for post-therapy surveillance without clinical, biochemical or radiographic evidence of disease remains controversial, primarily because of the potential for a disproportionate fraction of false-positive findings, potentially resulting in increasing cost without proven benefit from earlier PET detection of disease compared to standard surveillance methods. Large prospective studies are therefore needed to determine whether routine surveillance by PET is both cost effective and whether it results in meaningful changes in patient management and/or outcome. PMID- 17124071 TI - Biology, clinical course and management of nodular lymphocyte-predominant hodgkin lymphoma. AB - Nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) differs in histological and clinical presentation from classical Hodgkin lymphoma (cHL). The typical morphologic signs of NLPHL are atypical "lymphocytic and histiocytic" (L&H) cells, which are surrounded by a non-neoplastic nodular background of small lymphocytes of B-cell origin. The NLPHL cells are positive for CD45, CD19, CD20, CD22 and CD79a, but lack expression of CD15 and CD30, the typical markers for cHL. NLPHL patients are predominantly of male gender with a median age of 37 years. Patients often present in early stages (63%) and rarely have B-symptoms (9%). Treatment of NLPHL patients using standard Hodgkin lymphoma (HL) protocols leads to complete remission (CR) in more than 95% of patients. Survival and freedom from treatment failure (FFTF) are worse in advanced-stage patients than in early-stage patients. Thus, patients in advanced and in early stages with unfavorable risk factors are treated similarly to cHL patients. In contrast, patients with early-stage NLPHL without risk factors can be sufficiently treated with reduced intensity programs having less severe adverse effects. As a result, treatment of early NLPHL is less clearly defined, including radiotherapy in extended field (EF) or involved field (IF) technique, combined modality treatment, and, more recently, monoclonal antibody rituximab. Watch and wait strategy plays an important role in pediatric oncology to avoid adverse effects associated with therapy. PMID- 17124072 TI - Evolving view of the in-vivo kinetics of chronic lymphocytic leukemia B cells. AB - B-cell chronic lymphocytic leukemia (B-CLL) has long been considered a disease of "accumulation," due to a presumed defect in programmed cell death. Recent data, however, suggest that B-CLL cells are born at a normal to an accelerated rate, with the rate of proliferation varying among patients. In addition, differences in birth rates, activation state, and inducibility appear to exist among subpopulations of cells within individual leukemic clones. The extent to which such dissimilarities influence clinical course and outcome is still unclear. This review examines the evidence supporting the existence of a proliferative compartment in B-CLL and the role that proliferating cells might play in the progression and evolution of this disease. PMID- 17124073 TI - New prognostic markers in CLL. AB - The individual prognosis of patients with chronic lymphocytic leukemia (CLL) is extremely variable. Although clinical stages remain the basis for assessing prognosis in CLL, a number of biological markers, particularly serum markers, cytogenetic abnormalities, IgVH mutations, CD38 and ZAP-70 expression in leukemic cells offer important, independent prognostic information. Before being incorporated into daily practice, however, these markers require standardization and validation in large, prospective trials. Meanwhile, treatment of patients with CLL not included in clinical studies should be decided on the basis of classical NCI/CLL Working Group criteria. An important area of research in CLL prognostication is the identification of markers useful for predicting response to therapy and its duration. Among them, del(17p), reflecting P53 abnormalities, is particularly important. Also relevant is del(11q), which points to ATM defects. There is also some correlation between IgVH mutational status, ZAP-70 and CD38 expression and response to therapy and its duration, although these relationships need further investigation. Finally, there is increasing evidence that response to therapy, particularly in those cases in which minimal residual disease is eradicated, is associated with longer survival. PMID- 17124074 TI - Current and investigational therapies for patients with CLL. AB - Clinical and laboratory investigations are driving the rapid change in treatments for patients with chronic lymphocytic leukemia (CLL). Randomized trials have demonstrated superior activity for fludarabine combined with cyclophosphamide versus single-agent fludarabine or chlorambucil as initial treatment. Chemoimmunotherapy holds promise for further improvement and is being tested in randomized trials. New combinations and agents are being identified and tested. Eliminating minimal residual disease is a therapeutic endpoint that may prove to prolong survival and is also under investigation in prospective clinical trials. Work continues toward improving survival and potentially curing patients of this disease. PMID- 17124075 TI - Optimal use of prognostic factors in non-Hodgkin lymphoma. AB - The management of non-Hodgkin lymphoma is complicated by wide heterogeneity within recognized subtypes. Patients with supposedly similar diagnoses can have remarkably varied clinical presentations, molecular profiles and clinical outcomes. Reliable prognostic markers could allow the identification of patient subsets that may benefit from alternate approaches. Historically, a large number of clinical and molecular prognostic factors have been elucidated. However, the recent introduction of new therapies such as monoclonal antibodies has revolutionized treatment practices and greatly improved outcomes. This has called into question the value of previously recognized prognostic factors that need to be revalidated in the era of immunochemotherapy. It would appear that the commonly used clinical indices (IPI and FLIPI) retain predictive capacity, although they may have limited ability to identify a very poor outcome group. Currently there are no molecular markers that have been revalidated and shown to retain significance in the setting of current treatment practices for diffuse large B-cell lymphoma or follicular lymphoma. The biologic insights provided by molecular studies should allow for more targeted therapies to be developed, which will increase treatment choice and the possibility of tailored therapy in the future. It is imperative that future steps forward be made in the context of well designed clinical trials with prospective correlative studies of clinical and biologic markers. This will allow us to continuously assess outcome predictors in the context of treatment change and to rationally design tailored treatment algorithms. PMID- 17124076 TI - Controversies in follicular lymphoma: "who, what, when, where, and why?" (not necessarily in that order!). AB - Follicular lymphoma (FL) is the most common subtype of indolent lymphoma. Specific "facts" about FL that were generated by past research and have been passed down as dogma to a majority of practicing oncologists over the past 20 to 30 years that need to be revisited, include: (1) do not initiate therapy soon after diagnosis in asymptomatic, advanced-stage patients since it does not change outcome; (2) initiate therapy with single-agent oral alkylators when intervention needed and "save" more aggressive combination chemotherapy for "later" since the standard chemotherapy regimen used did not seem to impact survival; (3) FL is an incurable disease and palliation of symptoms was an acceptable approach to the expected pattern of repeated relapses; (4) transformation of FL is independent of the type or timing of therapies received by a patient; (5) median overall survival (OS) for FL patients is 8-10 years. Although the heterogeneity of FL will never change, we are developing the scientific tools to identify and better understand the biologic and genetic features associated with its clinical variability. In the current exciting era of targeted therapies (e.g., rituximab, radioimmunoconjugates) and novel treatment approaches demonstrating an improvement in treatment outcomes (e.g., disease-free survival and OS), our old beliefs and historically accepted dogma need to be retested and revitalized. The optimal combination(s) of old and new agents and the optimal timing of when to initiate and how to sequence specific therapies will require data from well designed clinical trials that should include important correlative laboratory studies. PMID- 17124077 TI - An update on therapy of primary central nervous system lymphoma. AB - The most important advance in primary central nervous system (CNS) lymphoma treatment has been the convincing data that high-dose methotrexate-based chemotherapy regimens improve survival compared to historical controls treated with radiotherapy alone. However, the optimal treatment approach is still unclear and therapy can be associated with long-term neurotoxicity. Current research focuses on maximizing survival while minimizing neurologic sequelae. PMID- 17124078 TI - Pathobiology of peripheral T-cell lymphomas. AB - Peripheral T-cell lymphomas (PTLs) are uncommon, accounting for fewer than 10% of all non-Hodgkin lymphomas. Success in therapy of the PTLs has lagged behind that of aggressive B-cell lymphomas, and most PTLs have a poor prognosis. The molecular pathogenesis of most PTLs is also poorly understood. In the WHO classification, clinical features, in conjunction with morphological and immunophenotypic criteria, are relied on to define most disease entities. Functionally, T-cell lymphomas are related to the two major arms of the immune system, the innate and adaptive immune systems. NK cells and T cells of the innate immune system recognize antigen in the absence of MHC antigens and are involved in mucosal immunity. The lymphomas derived from these cells often involve cutaneous and mucosal sites. The expression of cytotoxic molecules in these lymphomas may predispose to apoptosis by tumor cells and normal bystander cells. Hepatosplenic T-cell lymphoma is a systemic disease derived from functionally immature innate effector cells, most often of gammadelta T-cell origin. In contrast, most nodal T-cell lymphomas belong to the adaptive immune system. Angioimmunoblastic T-cell lymphoma (AILT) is mostly likely derived from follicular helper T-cells (T(FH)), a finding that explains many of its pathological and clinical features. Studies of these neoplasms may assist in further unraveling the functional diversity of their normal counterparts. PMID- 17124079 TI - Primary cutaneous T-cell lymphomas. AB - Primary cutaneous T-cell lymphomas (CTCLs) encompass a clinically and biologically heterogeneous group of non-Hodgkin lymphomas (NHLs) defined by clonal proliferation of skin-homing malignant T lymphocytes and natural killer cells. They account for up to 75% to 80% of all cutaneous lymphomas. The current WHO-EORTC classification of cutaneous lymphomas with primary cutaneous manifestations lists 13 entities. The most common subtypes-mycosis fungoides, Sezary syndrome, primary cutaneous anaplastic large cell lymphoma, and lymphomatoid papulosis-which represent approximately 95% of CTCLs, will be discussed in the following review. Each entity has unique biological characteristics and clinical course. Topical and/or systemic therapies are employed based on the stage of the disease and the tempo of progression. PMID- 17124080 TI - Therapy of peripheral T/NK neoplasms. AB - The mature T/natural killer (NK) lymphoma/leukemias represent 5-15% of all non Hodgkin lymphoma. These diseases have a geographic variation, with more nodal disease in North America and Europe, including peripheral T cell lymphomas, unspecified, anaplastic large cell lymphoma, and angioimmunoblastic T cell lymphoma; and more extranodal disease in Asia due to Epstein-Barr virus-related nasal NK/T lymphoma and human T-cell leukemia virus (HTLV)-1-associated adult T cell leukemia/lymphoma. The prognosis in most peripheral T/NK neoplasms is poor, with 5-year survival less than 30%. Progress has been slow due to the rarity of the diseases, geographic variation, relative chemoresistance, and lack of randomized trials. There is no consensus about optimal therapy in T/NK neoplasms, and recommendations are based on anecdotal reports, small series, and phase II trials. In this review, topics include the question of CHOP as standard therapy, prognostic factors, disease-adapted therapy, novel approaches, monoclonal antibody therapy, and stem cell transplantation. PMID- 17124081 TI - Initial therapy of multiple myeloma patients who are not candidates for stem cell transplantation. AB - Multiple myeloma patients deemed to not be candidates for high-dose therapy followed by stem cell rescue who nonetheless need chemotherapy have traditionally received an oral regimen combining melphalan and prednisone. With the advent of novel agents, however, such as immunomodulatory drugs and proteasome inhibitors that are active in the relapsed/refractory setting, there has been an impetus to incorporate these new options into front-line therapy. Several phase II studies have recently revealed that addition of either thalidomide, lenalidomide, or bortezomib to melphalan and prednisone increased the overall and complete response rates, albeit at the cost of some increased toxicity. Randomized phase III studies of melphalan and prednisone with thalidomide have already shown that, compared to melphalan and prednisone alone, the three-drug regimen prolonged time to progression and overall survival in this population, thereby defining a new standard of care. Moreover, our increasing knowledge of the molecular role that cytogenetic abnormalities play in the biology of multiple myeloma and our growing chemotherapeutic armamentarium are beginning to allow us to rationally select therapies based on these characteristics of each patient's disease. Such a risk- and molecular-adapted strategy to the therapy of multiple myeloma promises to revolutionize and personalize our care of these patients and bring us closer to a cure for this disease. PMID- 17124082 TI - Thrombotic complications of myeloma therapy. AB - Patients with multiple myeloma are at relatively high baseline risk of developing thromboembolic events (TEE), usually deep vein thromboses. There are numerous contributing factors, among them certain treatment regimens that include thalidomide or related compounds such as lenalidomide combined with glucocorticoids and/or cytotoxic chemotherapy. The risk of developing TEE appears to be particularly high when these immunomodulatory agents are combined with anthracyclines as treatment of newly-diagnosed disease. Up-front combinations including thalidomide plus pulse dexamethasone and/or alkylating agents are associated with an intermediate risk, whereas the same regimens for relapsed/refractory myeloma seem to be associated with the lowest risk. Several different thromboprophylaxis strategies have been effective in lowering the risk of developing clots: daily aspirin (81-325 mg/day), full-intensity warfarin (INR 2-3), and prophylactic enoxaparin (40 mg SQ daily). Low, fixed-dose warfarin may also reduce the risk of TEE, but the data on this are disputable. None of these TEE prevention strategies have been prospectively compared head-to-head, so the choice often reflects physician and/or patient preferences. The available evidence upon which one might make such a decision is reviewed here. PMID- 17124083 TI - Bisphosphonate complications including osteonecrosis of the jaw. AB - Bisphosphonate therapy has been incorporated in the standard management of patients with multiple myeloma-related bony disease. Although their efficacy in reducing skeletal related events is important in the supportive management of the myeloma patient, post-marketing experience with this class of agents, particularly the more potent intravenous agents pamidronate and zoledronic acid, have raised cautionary notes regarding the potential side effects of these agents. The focus of this session is to review the risk factors, incidence, prevention strategies and management of bisphosphonate-related osteonecrosis of the jaw. In addition, pathophysiology, incidence and monitoring for renal dysfunction during chronic therapy with these agents are reviewed. PMID- 17124084 TI - Of yeasts and hyphae: a hematologist's approach to antifungal therapy. AB - Improvements in anticancer treatments, the ability to modify myelosuppression profiles, greater duration and intensity of immunosuppression, and the variety of available antimicrobial therapies have influenced the spectrum of pathogens associated with invasive fungal infection complicating treatment of hematological malignancies and hematopoietic stem cell transplantation. The approaches to the management of these infections encompass strategies of prevention for all those at risk, preemptive therapy based upon surrogates of infection before the onset of clinical disease, empirical therapy for patients with clinical evidence of early disease, and directed or targeted therapy for infected patients with established disease. Chemoprophylaxis is effective if applied to the highest risk patients over the duration of the risk. Preemptive strategies, while promising, have yet to be validated and linked to reliably predictive non-microbiological diagnostic techniques. Empirical antifungal therapy, as it is currently applied, now seems questionable. Patients with probable or proven invasive fungal infection still have suboptimal outcomes despite the availability of promising anti-fungal agents. Strategies examining the concept of dose-intensity and combination regimens require careful study and cannot yet be regarded as an acceptable standard of practice. PMID- 17124085 TI - Viral infections in patients with hematological malignancies. AB - Viral infections are important causes of morbidity and mortality for patients with a hematological malignancy. However, the true incidence and consequences of viral infections for these patients who undergo conventional nontransplant therapy are poorly defined. The difference in incidence and outcome of viral infections among patient groups is wide, but dependent upon the intensity and duration of T-cell-mediated immune suppression. Infections caused by cytomegalovirus (CMV), herpes simplex virus (HSV), varicella-zoster virus (VZV), respiratory syncytial virus (RSV), parainfluenza viruses and influenza viruses have been intensely studied, yet newly recognized aspects of these viral infections including late CMV infection; the emergence of new viral pathogens (human herpesvirus-6, BK virus, adenovirus, and human metapneumovirus); the development of molecular diagnostic techniques, and the potential of new agents for viral prophylaxis (maribavir), or preemptive therapy (valganciclovir) form the basis of this review. Well-designed prospective studies are needed to better clarify the spectrum of these viral infections and develop effective prevention and treatment strategies. Yet the increased use of agents like alemtuzumab that induce profound T-cell depletion demands that we develop a better understanding of viral infections that occur in patients with hematological malignancy who receive nontransplant therapy. PMID- 17124086 TI - Viral hepatitis: manifestations and management strategy. AB - Viral hepatitis is the third most common cause of liver disease in allogeneic transplant recipients and causes significant morbidity and mortality. When treating patients with hematological malignancies, an emphasis should be placed on identification of patients at risk for viral hepatitis with appropriate screening. Initial screening serology should include anti-HCV, HBsAg, anti-HBs, and anti-HBc testing. When hepatitis B exposure has been documented, prophylaxis of viral reactivation for all HBsAg-positive patients with a nucleoside analogue should be implemented. HCV infection appears to have little short-term impact on survival after bone marrow transplantation, but is a risk factor for veno occlusive disease (VOD) and graft-versus-host disease (GVHD). In the long-term survivor, HCV infection can lead to significant morbidity and mortality due to the development of cirrhosis, decompensation, and liver cancer. Since effective antiviral therapies are available for both hepatitis B and C, routine screening and selected intervention is recommended once reactivation and disease recurrence is documented. In this chapter we will highlight the mechanisms of virus reactivation, clinical manifestations, and management strategies to minimize acute and chronic morbidity in this population. PMID- 17124087 TI - Outcomes with myeloid malignancies. AB - Reduced-intensity conditioning (RIC) regimens were initially introduced to provide allogeneic stem cell transplantation (HCT), a potentially curative procedure for myeloid malignancies, for patients who were not considered eligible for conventional myeloablative HCT either because of advanced age or excessive comorbidities. A variety of RIC regimens have been studied. The exact definition of RIC remains arbitrary and generally depends upon the perceived toxicity of a given regimen rather than the actual dose of chemotherapy or radiotherapy administered. In several published series, RIC regimens have demonstrated a reduction in non-relapse mortality (NRM), thereby accomplishing the initial goal of expanding the patient population eligible for this potentially curative procedure. Most retrospective studies performed to date have shown a decrease in NRM and an increase in relapse-related mortality with the use of RIC as opposed to conventional myeloablative HCT in myeloid malignancies. This appears to be particularly true for patients who are in relapse at the time of HCT. In contrast, patients who are in remission at time of HCT appear to have a reduction in NRM without a subsequent increase in relapse-related mortality. There is interest in applying RIC to younger patients and to patients with fewer comorbidities as they may have a reduction in NRM without a concomitant increase in relapse. Prospective multicenter studies are needed to define the optimal conditioning regimen, which is likely dependent upon a variety of disease specific and patient-specific factors. PMID- 17124088 TI - Reduced-intensity regimens in allogeneic stem-cell transplantation for non hodgkin lymphoma and chronic lymphocytic leukemia. AB - Autologous stem-cell transplantation is widely accepted as effective therapy for patients with relapsed aggressive B-cell non-Hodgkin lymphomas. Although 40-60% of younger patients with diffuse large cell lymphoma can expect to be cured, substantial numbers will experience a relapse. In addition, certain histologic subtypes are associated with particularly poor prognoses with combination chemotherapy alone (e.g., mantle cell lymphoma). Relatively few of these patients will experience long-term responses. Although other NHL subtypes are associated with more favorable prognoses in terms of overall survival, they are rarely cured (e.g., follicular lymphoma, chronic lymphocytic leukemia). Allogeneic transplantation has been increasingly utilized in patients with lymphoid malignancies but is associated with high toxicity. Recently, reduced-intensity conditioning regimens have shown encouraging results, attributed to graft-versus lymphoma effects. This article discusses changes in the way autologous and allogeneic transplants may be carried out in the future to treat patients with lymphoid malignancies. PMID- 17124089 TI - Reduced-intensity regimens in allogeneic hematopoietic stem cell transplantation for hemoglobinopathies. AB - The only well-established curative therapy for patients with hemoglobinopathies is allogeneic hematopoietic stem cell transplantation (HSCT), which, in the last 20 years, has been mainly performed from an HLA-matched, related donor, using bone marrow as source of hematopoietic progenitors. More recent studies indicate that HSCT from unrelated donors may offer results comparable to those obtained with HLA-identical family donors, provided that stringent criteria of compatibility are employed for selecting the donor. Cord blood transplantation was also suggested to be an equally effective, but safer, procedure than bone marrow transplantation, due to the lower incidence and severity of both acute and chronic graft-versus-host disease. In view of the early, as well as late, morbidity and mortality associated with conventional myeloablative transplantation in patients with hemoglobinopathies, it is not surprising that great interest and relevant expectations for patients with hemoglobinopathies have been raised by the introduction in the clinical practice of reduced intensity preparative regimens. However, few reports have demonstrated the feasibility of using reduced-intensity preparative regimens for successfully treating these patients and many treatment failures, mainly due to the lack of sustained donor engraftment, have been reported. Despite these limitations, some of the concepts obtained from the use of reduced intensity regimens, such as the substitution of fludarabine for cyclophosphamide, may be important to further improve the outcome of patients with hemoglobinopathies, especially of those with poor prognostic characteristics, given HSCT. PMID- 17124090 TI - ITP in the 21st century. AB - Immune (or idiopathic) thrombocytopenic purpura (ITP) is commonly encountered by the practicing hematologist. Clinical management decisions have traditionally been guided by individual training and past experience. Input from the literature has been more from observational reports of case series than from scientific results of hypothesis-driven research. Practice guidelines and several surveys of clinical hematology practice have highlighted important questions in the field, and in the past 5 to 10 years both clinical and laboratory investigations have produced valuable new information. Thrombopoietin levels are normal or only slightly increased in ITP, and stimulation of thrombopoiesis appears to be a promising new therapeutic approach in clinical trials. Chronic, refractory ITP in children or adults remains a challenge for the hematologist. It is this group that has the greatest risk of serious bleeding, particularly among the elderly. The anti-B-cell monoclonal antibody, anti-CD20, has shown benefit in phase I/II clinical trials in patients who had failed a number of previous therapeutic modalities. The standard for clinical research into therapy for ITP has become evidence-based medicine, and more prospective, randomized clinical trials are being completed by multi-institutional study groups. PMID- 17124091 TI - Think of HIT. AB - Heparin-induced thrombocytopenia, or HIT, can present in many ways, ranging from common-isolated thrombocytopenia, venous thromboembolism, acute limb ischemia-to less common but specific presentations-necrotizing skin lesions at heparin injection sites, post-bolus acute systemic reactions, and adrenal hemorrhagic necrosis (secondary to adrenal vein thrombosis). Many patients with HIT have mild or moderate thrombocytopenia: the median platelet count nadir is 60 x 10(9)/L, and ranges from 15 to 150 x 10(9)/L in 90% of patients, most of whom evince a 50% or greater fall in the platelet count. HIT that begins after stopping heparin ("delayed-onset HIT") is increasingly recognized. Factors influencing risk of HIT include type of heparin (unfractionated heparin > low-molecular-weight heparin), type of patient (surgical > medical), and gender (female > male). Since timely diagnosis and treatment of HIT may reduce the risk of adverse outcomes, this review focuses on those clinical circumstances that should prompt the clinician to "think of HIT." Coumarin anticoagulants such as warfarin are ineffective in acute HIT and can even be deleterious by predisposing to micro-thrombosis via protein C depletion (venous limb gangrene and skin necrosis syndromes). Thus, it is important to avoid or postpone coumarin while managing HIT hypercoagulability, focusing on agents that inhibit thrombin directly (lepirudin, argatroban) or that inhibit its generation (danaparoid, fondaparinux). Post-marketing experience suggests that standard dosing of lepirudin is too high; current recommendations are to avoid the initial lepirudin bolus and to begin with lower infusion rates, even in patients without overt renal dysfunction. PMID- 17124092 TI - Thrombotic thrombocytopenic purpura: a moving target. AB - Almost 80 years after Eli Moschcowitz published the first description of the disease, most patients with idiopathic thrombotic thrombocytopenic purpura (TTP) were found to have acquired autoantibody inhibitors of the ADAMTS13 metalloprotease. Plasma ADAMTS13 normally cleaves von Willebrand factor within nascent platelet-rich thrombi, and ADAMTS13 deficiency allows unchecked thrombus growth to cause microangiopathic hemolysis, thrombocytopenia, and tissue infarction. At present, ADAMTS13 deficiency with a high-titer inhibitor level appears to be associated with an increased risk of early death and subsequent relapse. Thus, acquired ADAMTS13 deficiency identifies a specific mechanism of TTP and is a potential biomarker of disease activity or risk. At present, two major clinical questions in the field may be summarized as follows. First, by emphasizing TTP caused by ADAMTS13 deficiency, are we in danger of neglecting other causes that should be treated with plasma exchange? Second, should we treat asymptomatic patients who have severe ADAMTS13 deficiency to prevent future disease, and if so, how? PMID- 17124093 TI - The role of immunomodulation in the management of factor VIII inhibitors. AB - Approximately 25% of persons with hemophilia A will have their treatment complicated by the development of anti-FVIII inhibitory antibodies. This adverse event requires the use of alternative hemostatic agents to treat bleeding and the consideration of a protocol to generate immunological tolerance to FVIII. The pathogenetic factors contributing to FVIII inhibitor generation include both patient- and concentrate-related characteristics. The FVIII genotype contributes to this risk as do other, less well defined, immunogenetic factors. The role of the type of FVIII concentrate as a precipitant for inhibitor generation appears to be less influential. Immunomodulatory management of FVIII inhibitors requires sustained and repeated exposure to FVIII through a variety of intravenous immune tolerance induction (ITI) protocols. Certain pre-ITI characteristics predict for the likelihood of success, most especially the pre-ITI anti-FVIII inhibitor titer. Currently, two major areas of debate remain unresolved with relation to the optimal form of ITI schedule. The best FVIII dose to generate FVIII tolerance is under investigation in an international prospective trial, while the issue of whether von Willebrand factor-containing concentrates may provide more powerful tolerizing effects remains open for further discussion. With a variety of ITI protocols, success rates of approximately 80% have been achieved with good-risk patients. In those that fail initial attempts at ITI, additional treatments using agents such rituximab are now being explored with further evidence of success in 60-80% of these salvage patients. Finally, several pre-clinical studies of innovative approaches to achieving FVIII tolerance suggest that combinations of immunomodulatory therapy may be of benefit in the future. PMID- 17124094 TI - Challenges in the therapeutic use of a "so-called" universal hemostatic agent: recombinant factor VIIa. AB - Recombinant factor VIIa (rFVIIa) was developed in the early 1990s to provide "bypassing" hemostatic therapy for hemophilia A and B patients with inhibitors. More recently, it has been licensed for use in patients with inherited deficiency of factor VII. Since it was licensed for use in hemophilia with inhibitors in the US, Europe, and other countries for these specific indications, it has been used selectively but in a wide array of clinical settings for uncontrolled hemorrhage in individuals without an inherited bleeding disorder. Many of these uses have been described in the medical literature as case reports or small, uncontrolled series. Several randomized clinical trials (RCT) for these "off-label" medical uses have been published in recent months and will serve as the focus of this review. In particular, a review of an RCT for spontaneous intracranial hemorrhage that has demonstrated clinical efficacy in reducing both mortality and volume of central nervous system hemorrhage will be offered. A brief discussion of hypothesized physiologic mechanisms of supraphysiologic doses of rFVIIa will introduce the clinical discussion of these broad off-label uses. Since rFVIIa is a very expensive therapy, possible strategies for optimizing its use in the these settings will be presented. PMID- 17124095 TI - Acquired factor VIII inhibitors: pathophysiology and treatment. AB - Hemophilia A is classically caused by a congenital deficiency of factor VIII, but an acquired form due to inhibitors to factor VIII (FVIII) typically presents later in life. Patients who develop such acquired factor VIII inhibitors may present with catastrophic bleeding episodes, despite having no prior history of a bleeding disorder. Though the disorder is rare, it is known to cause significant morbidity and mortality. This review will focus on what is currently known about acquired hemophilia A, its pathogenesis, its associated etiologies, and its treatment. PMID- 17124096 TI - Thrombosis and cancer: the role of screening for occult cancer and recognizing the underlying biological mechanisms. AB - The association between cancer and thrombosis is well recognized. What is not known, however, is the exact relationship between these two common medical conditions. Although the development of venous thromboembolism (VTE) in a patient with known cancer is the most common presentation, in some patients, VTE may precede the diagnosis of malignancy by many months. The variation in clinical presentation is likely due to the heterogeneous biology of different tumor types and also reflects the limitations of detection or available diagnostic methods. Accumulating evidence now suggests that critical oncogenic events may also trigger activation of the coagulation cascade, leading to a prothrombotic environment that not only manifests as venous thromboembolic disease but also promotes the growth and progression of the malignancy. This chapter will review the evidence for screening for occult malignancy in patients presenting with unprovoked or idiopathic thrombosis, briefly outline the known biological relationships between malignancy and thrombosis, and summarize the clinical data on the potential anticancer effects of low molecular weight heparins (LMWHs). PMID- 17124097 TI - Management of hereditary hypercoagulable disorders. AB - The clinical management of individuals with hereditary hypercoaguable disorders has evolved from initial broad recommendations of lifelong anticoagulation after first event of venous thromboembolism to a more intricate individualized risk benefit analysis as studies have begun to delineate the complexity of interactions of acquired and hereditary factors which determine the predilection to thrombosis. The contribution of thrombophilic disorders to risk of thrombotic complications of pregnancy, organ transplantation, central venous catheter and dialysis access placement have been increasingly recognized. The risk of thrombosis must be weighed against risk of long-term anticoagulation in patients with venous thromboembolism. Thrombophilia screening in select populations may enhance outcome. PMID- 17124098 TI - New anticoagulants. AB - Traditional anticoagulant drugs, including unfractionated heparin and warfarin, have several limitations. New anticoagulants have been developed that target a single coagulation factor and have predictable dose-response relationships. These include direct thrombin inhibitors and factor Xa inhibitors. Two parenteral direct thrombin inhibitors, lepirudin and argatroban, have FDA approval for the management of heparin-induced thrombocytopenia (HIT). Bivalirudin is a parenteral direct thrombin inhibitor that is licensed for patients undergoing percutaneous coronary interventions and for those with HIT who require percutaneous coronary interventions. Ximelagatran, an oral prodrug of the direct thrombin inhibitor melagatran, showed efficacy in the prevention and treatment of venous thromboembolism as well as stroke prevention in patients with atrial fibrillation. However, due to nonhematologic safety concerns, it did not receive FDA approval in the US. Fondaparinux is a synthetic pentasaccharide, which binds to antithrombin, thereby indirectly selectively inhibiting factor Xa. Fondaparinux demonstrated efficacy compared to low-molecular-weight heparin in randomized clinical trials and is FDA approved for the prevention and treatment of venous thromboembolism. The OASIS 5 trial in non-ST-segment elevation acute coronary syndromes recently demonstrated that the fondaparinux dose approved for prophylaxis of deep venous thrombosis is as efficacious with respect to ischemic outcomes as therapeutic doses of enoxaparin; fondaparinux, however, was associated with a substantial reduction in major bleeding at 9 days and mortality at 1 and 6 months. A number of oral direct factor Xa inhibitors as well as other oral direct thrombin inhibitors are in clinical development for the prevention and treatment of thrombosis; the current status of these anticoagulants is reviewed along with the challenges faced in designing pivotal clinical trials of these agents in comparison to existing anticoagulants. PMID- 17124099 TI - Perioperative evaluation of bleeding diathesis. AB - The differential diagnosis of a long APTT with a normal prothrombin time can be due to either a clotting factor deficiency or the presence of an inhibitor, which can be distinguished by using a plasma-mixing study. The various clotting factor deficiency states are reviewed. Clinical bleeding following cardiac bypass surgery due to acquired factor V and thrombin antibodies is also reviewed. PMID- 17124100 TI - Prevention of venous thromboembolism in high-risk patients. AB - The prevention of venous thromboembolism (VTE) in patients recovering from major trauma, spinal cord injury (SCI), or other critical illness is often challenging. These patient groups share a high risk for VTE, they often have at least a temporary high bleeding risk, and there are relatively few thromboprophylaxis trials specific to these populations. A systematic literature review has been conducted to summarize the risks and prevention of VTE in these three groups. It is concluded that routine thromboprophylaxis should be provided to major trauma, SCI and critical care patients based on an individual assessment of their thrombosis and bleeding risks. For patients at high risk for VTE, including those recovering from major trauma and SCI, prophylaxis with a low molecular weight heparin (LMWH) should commence as soon as hemostasis has been demonstrated. For critical care patients at lower thrombosis risk, either LMWH or low-dose heparin is recommended. For those with a very high risk of bleeding, mechanical prophylaxis should be instituted as early as possible and continued until pharmacologic prophylaxis can be initiated. The use of prophylactic inferior vena caval filters is strongly discouraged because their potential benefit has not been shown to outweigh the risks or substantial costs. Implementation of thromboprophylaxis in these patients requires a local commitment to this important patient safety priority as well as a highly functional delivery system, based on the use of pre-printed orders, computer prompts, regular audit and feedback, and ongoing quality improvement efforts. PMID- 17124101 TI - Pharmacogenetics-based coumarin therapy. AB - To reduce the risk of hemorrhage, experts advocate prescribing the anticipated therapeutic dose to patients who are beginning coumarin therapy, but until now there was no accurate way to estimate that dose. Using pharmacogenetics-based coumarin therapy, clinicians can now estimate the therapeutic dose by genotyping their patients for single nucleotide polymorphisms (SNPs) that affect coumarin metabolism or sensitivity. SNPs in the cytochrome P450 complex (CYP2C9) affect coumarin metabolism. Patients with either of two common variants, CYP2C9*2 or CYP2C9*3, metabolize coumarins slowly and are twice as likely to have a laboratory or clinical adverse event, unless their initial coumarin doses are reduced. SNPs in vitamin K epoxide reductase (VKORC1) correlate with coumarin sensitivity. Patients known to be homozygous for a common VKORC1 promoter polymorphism, -1639 G>A (also designated as VKOR 3673, haplotype A, or haplotype*2), should be started on lower coumarin doses than genotype GG patients. By providing an estimate of the therapeutic coumarin dose, pharmacogenetics-based therapy may improve the safety and effectiveness of coumarin therapy. PMID- 17124102 TI - Bleeding disorders in premenopausal women: (another) public health crisis for hematology? AB - Premenopausal women with bleeding disorders represent a major public health problem. Estimates suggest up to 20% of women with menorrhagia have an underlying bleeding disorder (corresponding to a prevalence of 1.5-4 million American women). Von Willebrand disease (VWD) is the most common bleeding disorder among women with menorrhagia, affecting up to 20% of such patients. Besides menorrhagia, important consequences of bleeding disorders in premenopausal women include iron deficiency anemia, miscarriage, postpartum bleeding, uterine bleeding and hysterectomy. These patients face many obstacles in achieving optimum care. Recognition is difficult as women may consider their symptoms "normal" and come to attention only after serious bleeding events. Symptoms of VWD may also overlap with benign conditions, primary providers may not suspect the diagnosis, and convenient hematologic input may be unavailable. Diagnosis is difficult as there is no single definitive test for VWD, and test results are variable, often being affected by extragenic factors, including stress, contraceptives, hormones, and pregnancy. Hemostatic treatment is limited by DDAVP tachyphylaxis, the lack of recombinant VWD concentrates, and the ineffectiveness of hormonal therapy, leading to unnecessary procedures and early hysterectomy. Finally, significant controversy exists regarding classification of type 1 VWD as a disease: given the overlap in symptoms and laboratory assays within the normal population, evaluation for those with VWD might be seen as identification of potential bleeding risk rather than detection of a disease. This symposium seeks to explore these issues in greater detail from the combined perspectives of the obstetrician-gynecologist and the hematologist to promote a better public health approach to this problem. PMID- 17124103 TI - Vaccine-enhanced donor lymphocyte infusion (veDLI). AB - Hematopoietic stem cell or bone marrow transplantation (HSCT/BMT) is curative in many cases of hemato-logical malignancy, but the post-transplant course is often complicated by delayed immune reconstitution that predisposes to opportunistic infections and disease recurrence. Furthermore, since HLA-matched donors cannot be found for almost half of all patients that would benefit from HSCT, donors mismatched at 2-3 HLA loci are increasingly being used, which is associated with elevated rates of opportunistic infections. Donor lymphocyte infusion (DLI) is a powerful and direct approach to improve post-transplant immune function. For example, DLI using enriched antiviral cytolytic effectors (CTLs) has been shown to reconstitute cellular immunity to cytomega-lovirus (CMV) and Epstein-Barr virus (EBV) and prevent viral disease following HSCT. However, because in vitro expansion and purification of CTLs is lengthy, labor-intensive, and costly, it is rarely used clinically to prevent and treat viral infections following HSCT. Active vaccination after allogeneic transplantation to stimulate in vivo expansion of donor and/or recipient CTLs has been proposed as an alternative method to rapidly reconstitute antiviral immunity, prevent viral disease, and reduce adverse sequelae of antiviral drugs. Fortunately, recent progress has been made in developing vaccines and methodologies that are both safe and effective when administered to immunocompromised HSCT recipients. PMID- 17124104 TI - Platelets: testing, dosing and the storage lesion--recent advances. AB - The demand for platelet transfusions continues to grow. Several complementary approaches that may help meet this demand in the future are reviewed. First, platelet bacterial testing is beginning to allow the extension of platelet storage beyond 5 days. Studies are also underway aimed at better preserving viability and function during ex vivo platelet storage: additive solutions and other approaches are being developed to try to negate the "platelet storage lesion." Finally, new approaches to dosing platelets may help extend the limited supply. PMID- 17124105 TI - Transfusion-related acute lung injury: an update. AB - Transfusion-associated acute lung injury (TRALI) has emerged as a leading cause of transfusion-related morbidity and mortality. TRALI is characterized by acute non-cardiogenic pulmonary edema and respiratory compromise in the setting of transfusion. The study of TRALI has been hampered by inadequate case definitions and an incomplete understanding of the pathologic mechanisms. Recent consensus conferences took an important first step by providing a framework for case definition. Recent advances in the understanding of the pathogenesis of TRALI have also occurred. TRALI has been primarily attributed to donor leukocyte antibodies that are thought to interact with recipient neutrophils, resulting in activation and aggregation in pulmonary capillaries, release of local biologic response modifiers causing capillary leak, and lung injury. An alternate mechanism termed the "two hit" or "neutrophil priming" hypothesis postulates that a pathway to neutrophil activation and aggregation can occur without leukocyte antibodies. A first event such as sepsis or trauma can induce pulmonary endothelial activation, release of cytokines, and priming of neutrophils. A subsequent second event such as exposure to lipids, cytokines or antibodies in a blood component would then cause activation of adherent neutrophils and a release of bioreactive molecules leading to lung injury. There are limited clinical and animal studies to support the "two hit" model. These proposed mechanisms are not mutually exclusive in that donor leukocyte antibody can be pathogenic in both models and have implications for new strategies to prevent TRALI. PMID- 17124107 TI - What is sports and exercise medicine? PMID- 17124108 TI - Personality correlates of physical activity: a review and meta-analysis. AB - This review aimed to combine the literature on major personality traits and physical activity alongside providing some meta-analytic summaries of the findings. Overall, 33 studies containing 35 independent samples, ranging from 1969 to 2006, met the inclusion criteria. Extraversion (r = 0.23), neuroticism (r = -0.11) and conscientiousness (r = 0.20) were identified as correlates of physical activity using random effects meta-analytic procedures correcting for sampling bias and attenuation of measurement error. The five-factor model traits of openness to experience/intellect and agreeableness, as well as Eysenck's psychoticism trait, were not associated with physical activity. Potential moderators of personality and physical activity relationships such as sex, age, culture/country, design and instrumentation were inconclusive given the small number of studies. Still, the existing evidence was suggestive that personality and physical activity relationships are relatively invariant to these factors. Studies examining personality and different physical activity modes suggested differences by traits such as extraversion, but more research is needed to make any conclusions. Future research using multivariate analyses, personality channelled physical activity interventions, longitudinal designs and objective physical activity measurement is recommended. PMID- 17124109 TI - High school rugby players' understanding of concussion and return to play guidelines. AB - OBJECTIVE: To document high school players' understanding and attitudes towards concussion return to play guidelines. METHODS: A questionnaire based survey was performed of national high school rugby players as to their knowledge of existing concussion return to play guidelines. RESULTS: A total of 600 male players were surveyed, and 477 responded (response rate 80%). Half (237/477) were aware of concussion guidelines, and 60% (288/477) identified the mandated stand down period that is part of the regulations governing rugby football. Players obtained their information primarily from: teachers/coaches (239 responses), medical personnel (200), and other players (116). Of those players who suspected that they had been concussed (296/477, 62%), only 66 returned to play after medical clearance. CONCLUSIONS AND IMPLICATIONS: This sample of high school players showed a limited knowledge of the concussion guidelines covering their sport, and even when concussed did not follow recommended protocols. This indicates the need for an increased focus on player education. PMID- 17124112 TI - Effect of convection on the measurement of thermophysical properties using levitated droplets. AB - This article discusses the recirculating convection in free droplets levitated by either the electromagnetic forces or the electrostatic forces, and its effect on the measurement of thermophysical properties. In an electromagnetically levitated droplet, strong internal flow results from the vortical Lorentz forces induced by the surrounding coils. These forces are also responsible for free surface deformation. In an electrostatically levitated droplet, however, internal convection originates from the thermal gradient and the electrostatic forces are responsible for surface deformation only. Mathematical models for predicting the convection in these droplets and the surface deformations caused by either the electromagnetic or electrostatic forces are discussed. Results show that internal convection, when present, can have a significant effect on the measured data for the purpose of determining the thermophysical properties. The deformed surface assumes essentially the same oval shape in microgravity for both electromagnetically and electrostatically levitated drops. The deformation mechanism, however, is different. Future research directions in this area are also discussed. PMID- 17124113 TI - Microgravity experiments on the effect of internal flow on solidification of Fe Cr-Ni stainless steels. AB - A new hypothesis has been developed to explain the effect of internal fluid flow on the lifetime of a metastable phase in solidifying Fe-Cr-Ni alloys. The hypothesis shows excellent agreement with available experimental results, but microgravity experiments are required for complete validation. Certain Fe-Cr-Ni stainless steel alloys solidify from an undercooled melt by a two-step process in which the metastable ferrite phase forms first followed by the stable austenite phase. Recent experiments using containerless processing techniques have shown that the lifetime of the metastable phase is strongly influenced by flow within the molten sample. Simulations using a commercial computational fluid dynamics (CFD) package, FIDAP, were performed to determine the time required for collision of dendrites and compared to experimental delay time. If the convective velocities are strong enough to bend the primary arms, then the secondary arms of adjacent dendrites can touch. The points of collision form low-angle boundaries and result in high-energy sites that can serve as nuclei for the transformation to the stable phase. It has been determined that the convective velocities in electrostatic levitation (ESL) are not strong enough to cause collision. However, in ground-based electromagnetic levitation (EML), the convective velocities are strong enough to cause the dendrites to deflect so that the secondary arms of adjacent dendrites collide. There is quantitative agreement between the numerically determined time to collision and the experimentally observed delay time in EML. The strong internal velocity due to convection within the EML samples is the reason for the observed difference in delay times between ESL and EML. Microgravity testing is essential because the significant change in nucleation behavior occurs between the ranges accessible by ground-based ESL and EML. Testing in microgravity using EML will permit a large range of internal convective velocities including those that are inaccessible in 1 g. PMID- 17124114 TI - Study on internal flow and surface deformation of large droplet levitated by ultrasonic wave. AB - It is expected that new materials will be manufactured with containerless processing under the microgravity environment in space. Under the microgravity environment, handling technology of molten metal is important for such processes. There are a lot of previous studies about droplet levitation technologies, including the use of acoustic waves, as the holding technology. However, experimental and analytical information about the relationship between surface deformation and internal flow of a large levitated droplet is still unknown. The purpose of this study is to experimentally investigate the large droplet behavior levitated by the acoustic wave field and its internal flow. To achieve this, first, numerical simulation is conducted to clarify the characteristics of acoustic wave field. Second, the levitation characteristic and the internal flow of the levitated droplet are investigated by the ultrasonic standing wave under normal gravity environment. Finally, the levitation characteristic and internal flow of levitated droplet are observed under microgravity in an aircraft to compare results with the experiment performed under the normal gravity environment. PMID- 17124115 TI - Containerless measurements of thermophysical properties of Zr54Ti8Cu20Al10Ni8. AB - High-temperature measurement and study of reactive materials can be difficult with conventional processing methods because contamination from the measuring apparatus and container walls can adversely affect measurements. Containerless processing techniques can be employed to isolate samples from their environment, reducing contamination. Benefits of containerless processing include reduction in heterogeneous nucleation sites, which in turn delays the onset of solidification and allows the study of meta-stable undercooled phases. However, property measurements must use noncontact methods as well. Fortunately, several optical based methods have been developed and successfully employed to measure thermophysical properties, including surface tension, viscosity, density, and thermal expansion. Combining these techniques with the electrostatic levitator (ESL) located at the NASA Marshall Space Flight Center (MSFC) has resulted in an excellent facility to perform containerless material studies which support microgravity flight projects. Currently, studies of the thermophysical properties of liquid quasi-crystal forming and related alloys ranging from superheated to deeply undercooled states are being done with this facility in support of the NASA-funded flight project Quasi-crystalline Undercooled Alloys for Space Investigation (QUASI). While the primary purpose of these measurements is to support planned flight experiments, they are also a desirable resource for future manufacturing considerations and for fundamental insight in the physics of icosahedral ordering in liquids and solids. Presented here is an overview of the contactless measuring methods for surface tension, viscosity, density, and thermal expansion applied to Zr54Ti8Cu20Al10Ni8, for the superheated and meta stable undercooled liquid phases, in support of QUASI. PMID- 17124116 TI - Noncontact thermophysical property measurement by levitation of a thin liquid disk. AB - The purpose of the current research program is to develop techniques for noncontact measurement of thermophysical properties of highly viscous liquids. The application would be for undercooled liquids that remain liquid even below the freezing point when suspended without a container. The approach being used here consists of carrying out thermocapillary flow and temperature measurements in a horizontally levitated, laser-heated thin glycerin disk. In a levitated state, the disk is flattened by an intense acoustic field. Such a disk has the advantage of a relatively low gravitational potential over the thickness, thus mitigating the buoyancy effects, and helping isolate the thermocapillary-driven flows. For the purpose of predicting the thermal properties from these measurements, it is necessary to develop a theoretical model of the thermal processes. Such a model has been developed, and, on the basis of the observed shape, the thickness is taken to be a minimum at the center with a gentle parabolic profile at both the top and the bottom surfaces. This minimum thickness is much smaller than the radius of disk drop and the ratio of thickness to radius becomes much less than unity. It is heated by laser beam in normal direction to the edge. A general three-dimensional momentum equation is transformed into a two variable vorticity equation. For the highly viscous liquid, a few millimeters in size, Stokes equations adequately describe the flow. Additional approximations are made by considering average flow properties over the disk thickness in a manner similar to lubrication theory. In the same way, the three-dimensional energy equation is averaged over the disk thickness. With convection boundary condition at the surfaces, we integrate a general three-dimensional energy equation to get an averaged two-dimensional energy equation that has convection terms, conduction terms, and additional source terms corresponding to a Biot number. A finite-difference numerical approach is used to solve these steady state governing equations in the cylindrical coordinate system. The calculations yield the temperature distribution and the thermally driven flow field. These results have been used to formulate a model that, in conjunction with experiments, has enabled the development of a method for the noncontact thermophysical property measurement of liquids. PMID- 17124117 TI - Transient dynamics and directional solidification in space platforms. AB - The impact of short impulsive forces on the semiconductor segregation patterns inside a generic mug Bridgman growth arrangement is discussed here with the help of a time-dependent 2D numerical scheme. Impulses have been applied parallel to the growth interface, equivalently, orthogonal to the external thermal gradient. The present results indicate that for each one of the three semiconductors considered, the reduced longitudinal and radial solid segregation are practically insensitive to the pulse shape, rectangular or half-sinusoidal. The reduced longitudinal segregation only depends on the growth velocity for constant g-dose impulses. The reduced radial segregation depends on both the interface growth velocity and very weakly on the activity time of the constant g-dose impulse externally applied. The thermal flux across the interface is shape-dependent and does not act synchronically with half-sinusoidal external impulses. Also, as before, this flux depends on the activity time and on the growth velocity for constant g-dose impulses. PMID- 17124118 TI - Effect of input diffusivity in an axisymmetric mass diffusivity model for liquid metals with an applied magnetic field. AB - Depending on the magnitude of the diffusivity, convective contamination may result in erroneous measurement of mass diffusivity even in microgravity. The effect of natural or buoyant convection is numerically investigated for liquid germanium with an axisymmetric model using two different magnetic field strengths and four heat transfer conditions. Since the reported input diffusivity value may vary due to inaccuracies in measurements, an order of magnitude lower and higher input diffusivity from the base value of D0 = 2 x 10(-4) cm2/sec was used to evaluate the behavior of liquid germanium in the presence of a magnetic field. From the numerical results, the temperature nonuniformity and the flow velocities in the liquid vary according to the input diffusivity (i.e., an order of magnitude lower and higher than the base cases). The heat transfer pattern is important in achieving a larger temperature nonuniformity in the liquid during the experiments. A stronger magnetic field can tolerate a higher temperature nonuniformity as expected. PMID- 17124119 TI - Developing quantitative, multiscale models for microgravity crystal growth. AB - Crystal growth conducted under microgravity conditions has had a profound impact on improving our understanding of melt crystal growth processes. Here, we present a brief history of microgravity crystal growth and discuss the development of appropriate models to interpret and optimize these growth experiments. The need for increased model realism and predictive capability demands new approaches for describing phenomena important at several disparate length scales. Of special importance is the ability to represent three-dimensional and transient continuum transport (flows, heat, and mass transfer), phase-change phenomena (thermodynamics and kinetics), and system design (such as furnace heat transfer during melt growth). An overview of mathematical models and numerical algorithms employed to represent such multiscale effects is presented. PMID- 17124120 TI - Effect of microgravity and magnetic field on the metallic and crystalline structure of magnetostrictive SmFe2 synthesized by unidirectional solidification. AB - The Sm-2Fe molten alloy with 1:2 molar ratio was unidirectionally solidified in both microgravity and normal gravity in concurrence with a magnetic flux (0-0.12 T). The compound SmFe2 was produced by the unidirectional solidification in microgravity with a magnetic flux of 0T and exhibited a lamellar microstructure. The average lamellar thickness was 30 mum and each lamella possessed a <111> crystallographic alignment along major axis aligned in the direction of cooling. Unidirectional solidification in microgravity with a magnetic field of 0.04 T produced crystalline SmFe2 and Fe phases. The microstructure of this product was lamellar with an average lamellar thickness of 17 mum and no crystalline alignment. Unidirectional solidification in microgravity with a magnetic flux ranging from 0.06 to 0.12 T and in normal gravity with a magnetic flux ranging from 0 to 0.12 T produced crystalline Sm2Fe17 and Fe. During unidirectional solidification in microgravity without a magnetic flux, few nucleation sites were formed and rapid crystal growth occurred, consequently forming large-grain SmFe2. The produced SmFe2 had a lamellar structure with a dominant <111> crystallographic alignment in the direction of cooling. Convection in the molten state and where a magnetic flux was present caused homogeneous nucleation, forming Sm2Fe17 with a disordered structure and crystalline alignment coinciding with the formation of the dendritic Fe. PMID- 17124121 TI - Convection effects on crystallinity in the growth of In0.3Ga0.7 as crystals by the traveling liquidus zone method. AB - The influence of convection in a melt on the crystallinity of the TLZ-grown In(0.3)Ga(0.7)As crystals has been investigated by growing crystals with various shapes and dimensions on the ground. No single crystals have been grown when the crystal diameter was 10 mm, but we were successful in growing single crystals by reducing crystal diameter to 2 mm. These results suggested the importance of suppressing convection in the melt during alloy crystal growth because constitutional supercooling tends to occur at the freezing interface or ahead of the interface by the segregation effect. Large area is required for substrate use in various applications. This requirement can be fulfilled by the crystal growth in microgravity because density difference-induced convection is suppressed in microgravity. Another means for suppressing convection without deteriorating area is plate-shape crystal growth with reduced thickness. The latter can be applied on the ground and we succeeded in growing single crystals of plate-shaped In(0.3)Ga(0.7)As by the traveling liquidus zone (TLZ) method. Dimensions of obtained single crystals were 10 mm in width and 2 mm in thickness and lengths ranged from 20 to 40 mm. Compositional uniformity was good and 0.3 +/- 0.02 in InAs mole fraction was achieved. PMID- 17124122 TI - Hypergravity as a crystallization tool. AB - The centrifugal increase of concentration is nondestructive, rapid, and simple technology. Therefore it is used to create a higher supersaturation that is required for crystal nucleation, as the one that is appropriate for the subsequent growth. Crystal nucleation is evoked in glass capillary tubes filled with protein solutions. The couple ferritin/ apoferritin is used as model proteins in the present article. Although differing in their masses the two (quasi) spherical molecules have exactly the same size and surface properties. Together with the temperature-independent solubility this makes them very convenient for our investigations. Decoupling nucleation and growth, for example, by means of hypergravity makes it possible to grow quasi-equidimensional crystals. The use of monodisperse crystalline forms of therapeutic agents can ensure constant time-release of protein-based medications. PMID- 17124123 TI - Macromolecular crystallization in microgravity generated by a superconducting magnet. AB - About 30% of the protein crystals grown in space yield better X-ray diffraction data than the best crystals grown on the earth. The microgravity environments provided by the application of an upward magnetic force constitute excellent candidates for simulating the microgravity conditions in space. Here, we describe a method to control effective gravity and formation of protein crystals in various levels of effective gravity. Since 2002, the stable and long-time durable microgravity generated by a convenient type of superconducting magnet has been available for protein crystal growth. For the first time, protein crystals, orthorhombic lysozyme, were grown at microgravity on the earth, and it was proved that this microgravity improved the crystal quality effectively and reproducibly. The present method always accompanies a strong magnetic field, and the magnetic field itself seems to improve crystal quality. Microgravity is not always effective for improving crystal quality. When we applied this microgravity to the formation of cubic porcine insulin and tetragonal lysozyme crystals, we observed no dependence of effective gravity on crystal quality. Thus, this kind of test will be useful for selecting promising proteins prior to the space experiments. Finally, the microgravity generated by the magnet is compared with that in space, considering the cost, the quality of microgravity, experimental convenience, etc., and the future use of this microgravity for macromolecular crystal growth is discussed. PMID- 17124124 TI - Is crystal growth under low supersaturations influenced by a tendency to a minimum of the surface-free energy? AB - The long-standing problem of face morphology is discussed. Special emphasis is put on macroscopically flat faces, whose growth, under low supersaturations, is driven by dislocations possessing some screw component. The most general case, where the crystal face is pierced by more than one screw dislocation, is considered. If performed under sufficiently low supersaturation, the growth leads to the formation of the face morphology corresponding to the minimum of the surface-free energy. The thermodynamic driving force for face flattening is the difference in the surface-free energy of the vicinal faces of the hillocks (emanating from screw dislocations) and that of a singular face, which can truncate the valley between the growth hillocks. The hillock slope gives the quantitative relation between energetics and kinetics. The result of the considerations is that the lower the supersaturation, the more important is the role of the surface-free energy in face flattening. Another factor, particularly under sufficiently low supersaturations, is the effective increase in the local supersaturation at the valley separating two growth hillocks. The reason is that the dislocation strain energy vanishes there. (This is most evident when two dislocations of opposite sign are considered.) Besides, the valley floors are concave regions on the crystal surface, where the building blocks are bound more strongly. Thus, the kinetic reason for face flattening is the relative preference for incorporation of atoms, arriving from the ambient phase, at the valley floor. Note that accelerated step annihilation in the valley floor should be a universal factor, which favors face flattening under any supersaturation. The amount of flattening in the growth situation is determined by the interplay between supersaturation and thermodynamics. PMID- 17124125 TI - Upside-down protein crystallization: designing microbatch experiments for microgravity. AB - The benefits of protein crystal growth in microgravity are well documented. The crystallization vessels currently employed for microgravity crystallization are far from optimal with regards to cost, sample volume, size, and ease of use. The use of microbatch experiments is a favorable alternative in each respect: 96 experiments of 0.5-2 microL volumes can be performed in a single microtiter tray measuring 5 x 8 cm and costing 1 pound sterling each. To date, the use of microbatch has not been pursued on account of concerns of oil leakage. To address this issue, a novel approach to microbatch crystallization experiments is described, where the microbatch plates are inverted throughout the duration of the experiment. The findings intimate the application of the microbatch method to space flight and the potential to drastically increase the output of microgravity crystallization research . PMID- 17124126 TI - Nucleation of protein crystals under the influence of solution shear flow. AB - Several recent theories and simulations have predicted that shear flow could enhance, or, conversely, suppress the nucleation of crystals from solution. Such modulations would offer a pathway for nucleation control and provide a novel explanation for numerous mysteries in nucleation research. For experimental tests of the effects of shear flow on protein crystal nucleation, we found that if a protein solution droplet of approximately 5 microL (2-3 mm diameter at base) is held on a hydrophobic substrate in an enclosed environment and in a quasi-uniform constant electric field of 2 to 6 kV cm(-1), a rotational flow with a maximum rate at the droplet top of approximately 10 microm s(-1) is induced. The shear rate varies from 10(-3) to 10(-1) s(-1). The likely mechanism of the rotational flow involves adsorption of the protein and amphiphylic buffer molecules on the air-water interface and their redistribution in the electric field, leading to nonuniform surface tension of the droplet and surface tension-driven flow. Observations of the number of nucleated crystals in 24- and 72-h experiments with the proteins ferritin, apoferritin, and lysozyme revealed that the crystals are typically nucleated at a certain radius of the droplet, that is, at a preferred shear rate. Variations of the rotational flow velocity resulted in suppression or enhancement of the total number of nucleated crystals of ferritin and apoferritin, while all solution flow rates were found to enhance lysozyme crystal nucleation. These observations show that shear flow may strongly affect nucleation, and that for some systems, an optimal flow velocity, leading to fastest nucleation, exists. Comparison with the predictions of theories and simulations suggest that the formation of ordered nuclei in a "normal" protein solution cannot be affected by such low shear rates. We conclude that the flow acts by helping or suppressing the formation of ordered nuclei within mesoscopic metastable dense liquid clusters. Such clusters were recently shown to exist in protein solutions and to constitute the first step in the nucleation mechanism of many protein and nonproteinsystems. PMID- 17124127 TI - An overview of challenges in modeling heat and mass transfer for living on Mars. AB - Engineering a life-support system for living on Mars requires the modeling of heat and mass transfer. This report describes the analysis of heat and mass transfer phenomena in a greenhouse dome, which is being designed as a pressurized life-support system for agricultural production on Mars. In this Martian greenhouse, solar energy will be converted into chemical energy in plant biomass. Agricultural products will be harvested for food and plant cultivation, and waste materials will be processed in a composting microbial ecosystem. Transpired water from plants will be condensed and recycled. In our thermal design and analysis for the Martian greenhouse, we addressed the question of whether temperature and pressure would be maintained in the appropriate range for humans as well as plants. Energy flow and material circulation should be controlled to provide an artificial ecological system on Mars. In our analysis, we assumed that the greenhouse would be maintained at a subatmospheric pressure under 1/3-G gravitational force with 1/2 solar light intensity on Earth. Convection of atmospheric gases will be induced inside the greenhouse, primarily by heating from sunlight. Microclimate (thermal and gas species structure) could be generated locally around plant bodies, which would affect gas transport. Potential effects of those environmental factors are discussed on the phenomena including plant growth and plant physiology and focusing on transport processes. Fire safety is a crucial issue and we evaluate its impact on the total gas pressure in the greenhouse dome. PMID- 17124128 TI - Heat and gas exchanges between plants and atmosphere under microgravity conditions. AB - Fundamental studies were conducted to develop a facility having an adequate air circulation system for growing healthy plants over a long term under microgravity conditions in space. To clarify the effects of gravity on heat and gas exchanges between plant leaves and the ambient air, surface temperatures and net photosynthetic rates of barley leaves were evaluated at gravity levels of 0.01, 1.0, and 2.0 g for 20 sec each during parabolic airplane flights. Thermal images were captured using infrared thermography at an air temperature of 22 degrees C, a relative humidity of 18%, and an irradiance of 260 W/m2. The net photosynthetic rates were determined by means of a chamber method with an infrared gas analyzer at an air temperature of 20 degrees C, a relative humidity of 50%, and photosynthetic photon fluxes (PPFDs) of 250 and 500 micromol/m2/sec. Mean leaf temperatures increased by 1.9 degrees C with decreasing gravity levels from 1.0 to 0.01 g and decreased by 0.6 degrees C with increasing gravity levels from 1.0 to 2.0 g. The increase in leaf temperatures was greater at the regions closer to the leaf tip and at most 2.5 degrees C over 20 sec as gravity decreased from 1.0 to 0.01 g. The net photosynthetic rate decreased by 20% with decreasing gravity levels from 1.0 to 0.01 g and increased by 10% with increasing gravity levels from 1.0 to 2.0 g at a PPFD of 500 micromol/m2/sec. The heat and gas exchanges between leaves and the ambient air were suppressed more at the lower gravity levels. The retardation would be caused by heat and gas transfers with less heat convection. Restricted free air convection under microgravity conditions in space would limit plant growth by retarding heat and gas exchanges between leaves and the ambient air. PMID- 17124129 TI - Gas embolism and surfactant-based intervention: implications for long-duration space-based activity. AB - Intravascular gas embolism can occur with decompression in space flight, and it commonly occurs during cardiac and vascular surgery. Intravascular bubbles may be deposited into any end organ such as the heart or the brain. Surface interactions between the bubble and the endothelial cells lining the vasculature result in serious impairment of blood flow and can lead to heart attack, stroke, or even death. Surfactant-based intervention is a novel treatment for gas embolism. Intravascular surfactant can adsorb onto the gas-liquid interface and compete with blood-borne macromolecules for interfacial occupancy. Surfactants can retard the progress of pathophysiological molecular and cellular events stimulated by the bubble surface, including endothelial cell injury and initiation of blood clotting. Bulk and surface transport of a surfactant to provide competition for interfacial occupancy is a therapeutic strategy because surfactant adsorption can dominate protein (or other macromolecule) adsorption. The presence of surfactant along the gas-liquid interface also induces variation in the interfacial tension, which in turn affects the blood flow and the bubble motion. We describe the interplay between biological transport processes and physiological events occurring and the cellular and molecular level in vascular gas embolization. Special consideration is given to modeling the transport and hydrodynamic interactions associated with surfactant-based intervention. PMID- 17124130 TI - Numerical modeling of the transport to an intravascular bubble in a tube with a soluble/insoluble surfactant. AB - Using a newly developed algorithm in conjunction with the front tracking scheme, we have evaluated the transport associated with a deformable bubble moving in a tube in the presence of a soluble or an insoluble surfactant. Such evaluations are useful to the understanding of gas embolism--a common syndrome for decompression sickness. Decompression sickness may be encountered in performing extravehicular activity during space exploration. The numerical evaluations indicate that as the location of the adsorptive interface gets closer to the vessel wall, the surfactant amount on the wall gets depleted. The implication is that the process by which a bubble occluding a vessel dislodges may depend both on the strength of the diffusivity of the surfactant and the adsorption process. More detailed study is needed to clarify this observation. The numerical results evaluated include Marangoni flow, which causes a bubble to propel out of its initial static location, and bubble motion in Poiseuille flow. The presence of a soluble/insoluble surfactant slows down the bubble motion. For identical surface concentrations of the surfactant, the effect of the presence of a soluble surfactant is more severe on the retardation of the bubble motion than that of an insoluble surfactant. PMID- 17124131 TI - A microgravity experiment of the on-orbit fluid transfer technique using swirl flow. AB - The cryogenic fluid transfer technique will prove useful for flexible and low cost space activities by prolonging the life cycle of satellites, orbital transfer vehicles, and orbital telescopes that employ cryogenic fluids, such as reactants, coolants, and propellants. Although NASA has conducted extensive research on this technique to date, a complicated mechanism is required to control the pressure in the receiver tank and avoid a large liquid loss by vaporization. We have proposed a novel fluid transfer method by using swirl flow combined with vapor condensation facilitated by spray cooling. This technique enables gas-liquid separation in microgravity and effectively facilitates vapor condensation without any special device like a mixer. In addition, since the incoming liquid flows along the tank wall, the tank wall would be cooled effectively, thereby minimizing the liquid loss due to vaporization. In this paper, the influence of the number of inlet points, fluid velocity at the inlet, fluid type, and boiling condition on swirl flow under microgravity conditions is investigated experimentally. The results indicated that the new fluid transfer technique using the swirl flow proposed by us is effective for cryogenic fluids that generally exhibit low surface tension and good wettability. In addition, it is possible to apply this technique to the real system because the swirl flow conditions are determined by the Froude number, which is dimensionless. Thus, the fundamental technique of fluid transfer by using the swirl flow under microgravity conditions was established. PMID- 17124132 TI - Hydrostatic compressibility phenomena: new opportunities for near-critical research in microgravity. AB - Peculiarities of the isothermal and isentropic equilibrium in highly compressible media with nonperfect state equations are discussed. Formulation of the adiabatic temperature gradient using Schwarzschild criterion and equilibrium equations for nonperfect gas are considered. Criterion for the onset of convection in the van der Waals (VDW) gas is examined using direct numerical modeling on the basis of equation for convection in compressible viscous nonperfect gas. Results of three dimensional modeling near-critical convection in the vicinity of the convection onset in space flight with quasi-steady microaccelerations are given. A proposal for the microgravity research of the near-critical convection is discussed. PMID- 17124133 TI - Axisymmetric surface oscillations in a cylindrical container with compensated gravity. AB - In this study the first mode of axisymmetric surface oscillations of a free liquid interface in a partly filled right circular cylinder under compensated gravity is investigated numerically. The situation is similar to a spacecraft that enters a ballistic flight at the end of thrust. A reorientation of the liquid of the free liquid surface toward the new equilibrium position takes place. The characteristics of this flow, such as the frequency and the damping, are governed by the behavior of the contact line. The investigations focus on the dependence on the contact line boundary condition, including contact angle variation, and on viscous effects. The effect on the natural frequency and the damping is considered. An analytical approximation for the frequency behavior for a frictionless liquid for a free and a fixed contact line condition is given. Further analytical approximations for the damping behavior in dependence on the contact angle and the Ohnesorge number are determined. The numerical results show very good agreement with experimental data. PMID- 17124134 TI - Fluctuations in diffusion processes in microgravity. AB - It has been shown recently that diffusion processes exhibit giant nonequilibrium fluctuations (NEFs). That is, the diffusing fronts display corrugations whose length scale ranges from the molecular to the macroscopic one. The amplitude of the NEF diverges following a power law behavior proportional to q(-4) (where q is the wave vector). However, fluctuations of wave number smaller than a critical "rolloff" wave vector are quenched by the presence of gravity. It is therefore expected that in microgravity conditions, the amplitude of the NEF should be boosted by the absence of the buoyancy-driven restoring force. This may affect any diffusion process performed in microgravity, such as the crystallization of a protein solution induced by the diffusion of a salt buffer. The aim of GRADFLEX (GRAdient-Driven FLuctuation EXperiment), a joint project of ESA and NASA, is to investigate the presence of NEFs arising in a diffusion process under microgravity conditions. The project consists of two experiments. One is carried out by UNIMI (University of Milan) and INFM (Istituto Nazionale per la Fisica della Materia) and is focused on NEF in a concentration diffusion process. The other experiment is performed by UCSB (University of California at Santa Barbara) concerning temperature NEF in a simple fluid. In the UNIMI part of the GRADFLEX experimental setup, NEFs are induced in a binary mixture by means of the Soret effect. The diagnostic method is an all-optical quantitative shadowgraph technique. The power spectrum of the induced NEFs is obtained by the processing of the shadowgraph images. A detailed description of the experimental apparatus as well as the ground-based experimental results is presented here for the UNIMI INFM experiment. The GRADFLEX payload is scheduled to fly on the FOTON M3 capsule in April 2007. PMID- 17124135 TI - Effect of gravity on the dynamics of nonequilibrium fluctuations in a free diffusion experiment. AB - Diffusion is commonly believed to be a homogeneous process at the mesoscopic scale, being driven only by the random walk of fluid molecules. On the contrary, very large amplitude, long wavelength fluctuations always accompany diffusive processes. In the presence of gravity, fluctuations in a fluid containing a stabilizing gradient are affected by two different processes: diffusion, which relaxes them, and the buoyancy force, which quenches them. These phenomena affect both the overall amplitude of fluctuations and their time dependence. For the case of free diffusion, the time-correlation function of the concentration fluctuations is predicted to exhibit an exponential decay with correlation time depending on the wave vector q. For large wave vector fluctuations, diffusion dominates, and the correlation time is predicted to be 1 / (Dq2). For small wave vector fluctuations, gravitational forces have time to play a significant role, and the correlation time is predicted to be proportional to q2. The effects of gravity and diffusion are comparable for a critical wave vector q(c) determined by fluid properties and gravity. We have utilized a quantitative dynamic shadowgraph technique to obtain the temporal correlation function of a mixture of LUDOX(R) TMA and water undergoing free diffusion. This technique allows one to simultaneously measure correlation functions achieving good statistics for a number of different wave vectors in a single measurement. Wave vectors as small as 70 cm(-1) have been investigated, which is very difficult to achieve with ordinary dynamic light-scattering techniques. We present results on the transition from the diffusive decay of fluctuations to the regime in which gravity is dominant. PMID- 17124136 TI - Preflight diffusion experiments on liquid metals under 1g conditions for the FOTON-M2 mission. AB - We measured diffusion of up to 5% Ag in liquid Pb, using the Foton shear cell (designed and used for the FOTON-M2 mission). Since the density change of the alloy with composition has been reported to be very small but is not reliably known, we performed two arrangements of layering: (a) with PbAg on top and Pb below and (b) with Pb on top and PbAg below. The Berlin group did interdiffusion experiments from a "thick layer" of PbAg into "semi-infinite" Pb and the Karlsruhe group did the typical "interdiffusion" experiments with a diffusion couple of PbAg versus Pb. The results of both groups were equivalent, showing much lower D values for arrangement (b) than for (a). The lower values were even definitely lower than reference microgravity values from MIR/MIM and could be fitted by D = AT(n) with n about 2.2. We discuss the stability/instability by density layering depending on the arrangement. PMID- 17124137 TI - Double-layer thermocapillary convection in a differentially heated cavity. AB - Many materials-processing applications such as crystal growth from the melt involve thermocapillary flows that can affect the quality of the final product, particularly under microgravity conditions where the influence of buoyancy-driven convection is minimized. When the melt contains volatile components, as in the production of III-V semiconductor crystals, it is often encapsulated in a low melting point amorphous molten glass phase such as boron oxide or pyrolytic boron nitride in order to prevent evaporation of the volatile components. The addition of the encapsulant layer and the melt-encapsulant interface in such cases can alter the thermocapillary flow in the melt. In this study, thermocapillary convection within a differentially heated rectangular cavity containing two immiscible liquid layers is considered in the absence of gravity. Domain mapping is used in conjunction with a finite difference scheme on a staggered grid to solve for the temperature and flow fields. The melt-encapsulant and the air encapsulant interfaces are allowed to deform, with the contact lines pinned on the solid boundaries. The computed flow fields are compared to the corresponding results for a cavity with a rigid top surface. The presence of a free surface at the top leads to increased convection in the encapsulant phase while suppressing the thermocapillary flow in the melt phase. The flow pattern in the encapsulated layer is strongly dependent on the viscosity of the encapsulant layer. The intensity of the thermocapillary flow within the melt is significantly reduced as the viscosity of the encapsulant layer is increased. However, for a higher encapsulant viscosity, the retarding effect of the free top surface on thermocapillary convection in the melt is weakened. PMID- 17124138 TI - An electrowetting microvalve: numerical simulation. AB - Numerical simulation of a zero-leakage microvalve is investigated where a liquid droplet is used as a gate to regulate the flow in a T junction. The droplet gate is activated by changing its surface tension via an applied electric field. Numerical simulation of the droplet actuation is considered where the effect of electrowetting is imposed in the form of a modified boundary condition at the contact line. Numerical simulation is used to predict the droplet behavior and to design the valve. It is found that the pressure breakdown of the microvalve is significantly affected by the geometry of the T junction corners. It is expected that such a microvalve design will improve the sensitivity and performance of a wide variety of microfluidic devices. PMID- 17124139 TI - Numerical simulation of the spontaneous penetration of liquids into cylindrical capillaries. AB - Dynamics of spontaneous capillary penetration of a liquid into a cylindrical pore is studied numerically over the entire duration of an experiment, including the initial stages of penetration during which inertial effects are dominant. Partial slip in the vicinity of the moving contact-line is allowed by using an empirical constitutive relation between the dynamic contact angle and contact line speed in order to avoid the stress singularity arising from the presence of the moving contact line on the solid wall. A finite-difference scheme on a staggered body fitted grid is used to solve for the time-dependent flow field and to determine the time evolution of the shape of the advancing meniscus. The results of dynamic simulations of capillary rise under both normal and microgravity conditions are compared with the reported experimental observations. The simulation results are found to be in good agreement with the experimental measurements in both cases. Numerical simulations capture the different flow regimes identified in previous studies of spontaneous capillary penetration. The velocity dependence of the dynamic contact line is found to have a significant effect on kinetics of wetting in the intermediate-time flow regime, wherein the capillary force is balanced by convective losses. PMID- 17124140 TI - Flow rate limitation in open capillary channel flows. AB - This paper reports the experimental and theoretical investigations of forced liquid flows through open capillary channels under reduced gravity conditions. An open capillary channel is a structure that establishes a liquid flow path at low Bond numbers, when the capillary pressure caused by the surface tension force dominates in comparison to the hydrostatic pressure induced by gravitational or residual accelerations. In case of steady flow through the channel, the capillary pressure of the free surface balances the pressure difference between the liquid and the surrounding constant-pressure gas phase. Because of convective and viscous momentum transport, the pressure along the flow path decreases and causes the free surface to bend inward. The maximum flow rate is achieved when the free surface collapses and gas ingestion occurs at the outlet. This critical flow rate depends on the geometry of the channel and the properties of the liquid. In this paper we present a comparison of the theoretical and experimental critical flow rates and surface profiles for convective dominated flows. For the prediction of the critical flow rate a one-dimensional theoretical model taking into account the entrance pressure loss and the frictional pressure loss in the channel is developed. PMID- 17124141 TI - Analysis of heat and mass transfer during condensation over a porous substrate. AB - Condensing heat exchangers are important in many space applications for thermal and humidity control systems. The International Space Station uses a cooled fin surface to condense moisture from humid air that is blown over it. The condensate and the air are "slurped" into a system that separates air and water by centrifugal forces. The use of a cooled porous substrate is an attractive alternative to the fin where condensation and liquid/gas separation can be achieved in a single step. We analyze the heat and mass transfer during condensation of moisture from flowing air over such a cooled, flat, porous substrate. A fully developed regime is investigated for coupled mass, momentum and energy transport in the gas phase, and momentum and energy transport in the condensate layer on the porous substrate and through the porous medium. PMID- 17124142 TI - Direct computational simulations and experiments for film condensation inside tubes and channels. AB - This article presents accurate numerical solutions of the full 2D governing equations for steady and unsteady laminar/laminar internal condensing flows of pure vapor (FC-72 and R-113) inside a vertical tube and a channel. The film condensation is on the inside wall of a tube or one of the walls of a channel (the lower wall in case of a downward sloping channel). Both experiments and simulations find that exit condition specifications are important. The computations are able to predict whether or not a steady flow exists with a well defined and steady natural exit condition. If well-defined natural steady/quasi steady flows exist-as is shown to be the case for gravity-dominated or strong shear-dominated condensate flows that remain parabolic up to the exit location the computations are able to predict both the natural exit condition and any point of transition (from stable to unstable or smooth to wavy behavior) that may exist within this zone. Compared to gravity-driven cases, shear-driven cases (zero gravity or horizontal cases) tend to destabilize easily. It is found that only for gravity-driven cases interfacial waves are able to cause a concurrent enhancement in heat transfer rates along with an enhancement in interfacial shear. Also it is found that this enhancement in interfacial wave energy is significant if the condensing surface noise is in resonance with the intrinsic waves. PMID- 17124143 TI - Coalescence of bubbles translating through a tube. AB - The results of an experimental study of the interaction and coalescence of two air bubbles translating in a cylindrical tube are presented. Both pressure- and buoyancy-driven motion of the two bubbles in a Newtonian suspending fluid within the tube are considered. The close approach of the two bubbles is examined using image analysis, and measurements of the coalescence time are reported for various bubble size ratios and capillary numbers. For pressure-driven motion of bubbles, coalescence is found to occur in an axisymmetric configuration for all bubble size ratios considered in the experiments. For buoyancy-driven motion, on the other hand, the disturbance flow behind the leading bubble causes the trailing bubble to move radially out toward the tube wall when the trailing bubble size becomes very small compared to the size of the leading bubble. In that case, coalescence occurs in a nonaxisymmetric configuration, with a time scale for coalescence that is substantially larger than that for coalescence in the axisymmetric configuration. When the imposed flow is in the direction of the buoyancy force, coalescence time is independent of bubble size ratio, and decreases as the capillary number increases. Experimental measurements of the radius of the thin liquid film separating the two bubbles are used in conjunction with a simple film drainage model to predict the dependence of the coalescence time on the bubble size ratio. PMID- 17124144 TI - Heat transfer enhancement by electric fields in several heat exchange regimes. AB - With the present article, the authors summarize over 15 years of work dedicated to studying the effects of the electrical and the gravitational force fields on two-phase and single-phase thermofluid dynamics. Results obtained on several microgravity platforms are presented and the role played by the electric field in the different heat transfer processes is analyzed. In particular, the regimes of nucleate boiling and film boiling are treated. Also, the parameters controlling the liquid-vapor interface instability and the main consequences regarding the critical heat flux (CHF) are outlined. In the final section, the promising technique of ion injection for efficient heat transfer enhancement in single phase liquids is described, together with the main results obtained under different flow regimes and geometries. The following dielectrics were compared: R113, Vertrel XF, and FC-72 for the pool boiling research; again FC-72 and HFE 7100 for the single-phase convection heat transfer experiments. PMID- 17124145 TI - Using large electric fields to control transport in microgravity. AB - Transport control by large electric fields in microgravity may be subdivided according to whether the charge carriers are flame ions, ions produced by corona discharges, or electrically charged particles. Using electric fields to induce and direct convection through the drag exercised on neutral gas by ions and to manipulate dispersions and trajectories of electrically charged droplets and particles is especially effective in the absence of Earth's gravity. We have explored applications associated with each of these, and this review collects and summarizes briefly the principal findings of our research, which is scattered widely over the literature of combustion, electrostatics, and experimental science. PMID- 17124146 TI - Canceling buoyancy of gaseous fuel flames in a gravitational environment using an ion-driven wind. AB - Electric fields applied to combustion plasmas can be used to manipulate the thermofluid flow field to reduce buoyant forces and, hence, convection in locations near and within the flame. The resulting flow field is similar to that which is obtained in microgravity. Previous work has shown that buoyancy is modified in a non-premixed methane-air capillary flame when it burns in a capillary-to-plane configuration and an electric field is applied, and that regions of neutral or microbuoyancy exist, as indicated by the examined temperature and oxidizer profiles. The aim of this article is to examine in more detail this microbuoyancy condition and the coupling between the ion wind and resulting thermofluid flow field. To this end, the voltage-current characteristics (VCC) of CH4, C2H2, C2H4, C2H6, and C3H8 are measured and compared. Soot generated in the C2H(X) and propane flames lead to a hysteresis in the VCC curve whereby increased sooting leads to lower ion currents at constant flow rates and applied potentials. Buoyancy regimes for these flames in this configuration are determined. Methane can achieve the highest flow rate without sooting at the microbuoyant condition, and does not exhibit hysteresis in the VCC for the flow rates examined here. Furthermore, in this geometry, the microbuoyant condition for methane is found to coincide with ion current saturation when the capillary-to-plane distance is varied. These results allow for several simplifications to be made when modeling the flame at these conditions: the imposition of a spherical flame boundary with known ion current, and negligible recombination in the domain. PMID- 17124147 TI - Augmentation of heat transfer on the downward surface of a heated plate by ion injection. AB - The heat transfer characteristics of an electrohydrodynamically induced submerged impinging liquid flow are studied in a point-plane configuration. The working fluid, HFE-7100, is a weakly polar dielectric. Thermofluid-dynamic conditions as close as possible to the microgravity ones are obtained by heating a confined, downward-facing plate. Heat transfer coefficients on the plate can be stably enhanced by over 10 times, with negligible additional work, by means of the ion injection technique. PMID- 17124148 TI - Numerical computation on magnetothermal air jet in gravitational and nongravitational fields. AB - Two-dimensional numerical computations were carried out in order to elucidate the effect of a Kelvin force on the air in two coaxial circular pipes with open ends under both gravitational and nongravitational fields. The outer pipe with open ends corresponds to the bore space of the superconducting magnet. The inner pipe with open ends is assumed to be placed inside this bore space. The sidewall of the outer pipe is cooled isothermally. The central region of the inner pipe is heated isothermally and the other region is thermally insulated. The magnetic gradient that was produced by the electric current circulating within the circular electric coil was applied to air in two coaxial circular pipes with a thermal gradient. Moreover, the present numerical computations were carried out by changing the relative positions of the circular electric coil and the inner pipe. In both gravitational and nongravitational fields, when the circular electric coil was placed at the end of the heated region of the inner pipe, the Kelvin force was produced in the inner pipe and the magnetothermal air jet was created. As a result, the hotter air rapidly flowed out from the end of the inner pipe. These phenomena could be successfully explained by considering the temperature dependence of the mass magnetic susceptibility of air according to Curie's law. PMID- 17124149 TI - Some parameter boundaries governing microgravity pool boiling modes. AB - Pool boiling experiments were conducted in microgravity on five space shuttle flights, using a flat plate heater consisting of a semitransparent thin gold film deposited on a quartz substrate that also acted as a resistance thermometer. The test fluid was R-113, and the vapor bubble behavior at the heater surface was photographed from beneath as well as from the side. Each flight consisted of a matrix of three levels of imposed heat flux and three levels of initial bulk liquid subcooling. In many of the total of 45 experiments, steady nucleate boiling was observed from 16-mm movie films, where a large vapor bubble formed and remained slightly removed from the heater surface, with small vapor bubbles growing on the heater surface, and on contact coalescing with the large bubble. Computations of the forces associated with the momentum transfer in this process, which counters the Marangoni convection effects tending to impel the large bubble toward the heater surface, have been completed for all cases where applicable. The modes of pool boiling observed with successive increases in levels of heat flux in microgravity are categorized as: (i) minimum or incipient nucleate boiling; (ii) nucleate boiling with vigorous motion of the bubbles adjacent and parallel to the heater surface, impelled by Marangoni convection effects; (iii) nucleate boiling followed by coalescence with a neighboring large vapor bubble; (iv) partial dryout of the heater surface, in parallel with nucleate boiling; (v) complete dryout. The boundaries between these modes are delineated graphically as a function of the imposed heat flux and initial bulk liquid subcooling, together with the levels of the forces holding the large bubbles, acting as vapor reservoirs, away from the heater surface for the steady nucleate boiling mode. PMID- 17124150 TI - Self-rewetting fluids: beneficial aqueous solutions. AB - This paper reviews a series of my heat transfer studies of on the so-called "self rewetting" fluids, in which the Marangoni effect due to concentration gradient and the thermocapillary flow cooperatively result in a strong liquid inflow at the nucleation sites in the course of boiling. First of all, the background of the study and our first attempt at heat transfer enhancement in microgravity boiling are introduced. Succeeding fundamental vapor bubble experiments are then briefly depicted. The current progress for applications to space and terrestrial thermal management devices, the main topic of the paper, is described in more detail. PMID- 17124151 TI - Structure of high-performance evaporators for space application. AB - A new structure of cold plates, where an unheated auxiliary channel is installed to supply liquid directly to the bottom of coalesced flattened bubbles in a narrow heated channel, is tested to investigate the increase in critical heat flux. Assuming the application to the laser solar power system, a large heating surface with a length of 150 mm in the flow direction is employed, and a narrow channel structure is adopted to reduce the size of cold plates, where the gap sizes are selected as 5 mm and 2 mm. Experiments are performed for water as a test liquid at inlet subcooling of 15 K under near atmospheric pressure. Inlet liquid velocity is varied from 0.065 m/s to 0.6m/s for the upward flow on ground. A value of critical heat flux of 2.2 x 10(6) W/m2 is obtained for 5-mm gap size at the inlet velocity of 0.2 m/s. At low liquid flow rate, the structure realizes the CHF values larger by 2.5 times than those for the normal heated channel without additional liquid supply. A new method to evaluate the performance of cold plates is proposed to take account of the variation in the size of heating surface, inlet liquid velocity, and subcooling that influence the CHF values. The validity of the proposed structure of the cold plate for the increase in critical heat flux is confirmed. PMID- 17124152 TI - Probiotics, prebiotics, synbiotics: ecological treatment for inflammatory bowel disease? PMID- 17124153 TI - Genes and environment in irritable bowel syndrome: one step forward. PMID- 17124154 TI - Metachronous tumour development in the pancreas. PMID- 17124156 TI - An unusual cause of abdominal distension. PMID- 17124157 TI - Unusual presentation of digestive tumour. PMID- 17124161 TI - Predictive value of microsatellite instability for benefit from adjuvant fluorouracil chemotherapy in colorectal cancer. PMID- 17124160 TI - Molecular basis of Barrett's oesophagus and oesophageal adenocarcinoma. PMID- 17124162 TI - Prospective evaluation of fluorouracil chemotherapy based on the genetic makeup of colorectal cancer. PMID- 17124163 TI - Paraoxonase 1, 2 and 3 DNA variants and susceptibility to childhood inflammatory bowel disease. PMID- 17124164 TI - British Society of Gastroenterology guidelines for the diagnosis of Barrett's oesophagus: are we casting the net too wide? PMID- 17124165 TI - Effect of bowel preparation and colonoscopy on post-procedure intestinal microbiota composition. PMID- 17124166 TI - Dynamic assembly of end-joining complexes requires interaction between Ku70/80 and XRCC4. AB - DNA double-strand break (DSB) repair by nonhomologous end joining (NHEJ) requires the assembly of several proteins on DNA ends. Although biochemical studies have elucidated several aspects of the NHEJ reaction mechanism, much less is known about NHEJ in living cells, mainly because of the inability to visualize NHEJ repair proteins at DNA damage. Here we provide evidence that a pulsed near IR laser can produce DSBs without any visible alterations in the nucleus, and we show that NHEJ proteins accumulate in the irradiated areas. The levels of DSBs and Ku accumulation diminished in time, showing that this approach allows us to study DNA repair kinetics in vivo. Remarkably, the Ku heterodimers on DNA ends were in dynamic equilibrium with Ku70/80 in solution, showing that NHEJ complex assembly is reversible. Accumulation of XRCC4/ligase IV on DSBs depended on the presence of Ku70/80, but not DNA-PK(CS). We detected a direct interaction between Ku70 and XRCC4 that could explain these requirements. Our results suggest that this assembly constitutes the core of the NHEJ reaction and that XRCC4 may serve as a flexible tether between Ku70/80 and ligase IV. PMID- 17124167 TI - Crystal structure of pi initiator protein-iteron complex of plasmid R6K: implications for initiation of plasmid DNA replication. AB - We have determined the crystal structure of a monomeric biologically active form of the pi initiator protein of plasmid R6K as a complex with a single copy of its cognate DNA-binding site (iteron) at 3.1-A resolution. The initiator belongs to the family of winged helix type of proteins. The structure reveals that the protein contacts the iteron DNA at two primary recognition helices, namely the C terminal alpha4' and the N-terminal alpha4 helices, that recognize the 5' half and the 3' half of the 22-bp iteron, respectively. The base-amino acid contacts are all located in alpha4', whereas the alpha4 helix and its vicinity mainly contact the phosphate groups of the iteron. Mutational analyses show that the contacts of both recognition helices with DNA are necessary for iteron binding and replication initiation. Considerations of a large number of site-directed mutations reveal that two distinct regions, namely alpha2 and alpha5 and its vicinity, are required for DNA looping and initiator dimerization, respectively. Further analysis of mutant forms of pi revealed the possible domain that interacts with the DnaB helicase. Thus, the structure-function analysis presented illuminates aspects of initiation mechanism of R6K and its control. PMID- 17124168 TI - Mitochondrial deoxynucleotide pool sizes in mouse liver and evidence for a transport mechanism for thymidine monophosphate. AB - Dividing cultured cells contain much larger pools of the four dNTPs than resting cells. In both cases the sizes of the individual pools are only moderately different. The same applies to mitochondrial (mt) pools of cultured cells. Song et al. [Song S, Pursell ZF, Copeland WC, Longley MJ, Kunkel TA, Mathews CK (2005) Proc Natl Acad Sci USA 102:4990-4995] reported that mt pools of rat tissues instead are highly asymmetric, with the dGTP pool in some cases being several hundred-fold larger than the dTTP pool, and suggested that the asymmetry contributes to increased mutagenesis during mt DNA replication. We have now investigated this discrepancy and determined the size of each dNTP pool in mouse liver mitochondria. We found large variations in pool sizes that closely followed variations in the ATP pool and depended on the length of time spent in the preparation of mitochondria. The proportion between dNTPs was in all cases without major asymmetries and similar to those found earlier in cultured resting cells. We also investigated the import and export of thymidine phosphates in mouse liver mitochondria and provide evidence for a rapid, highly selective, and saturable import of dTMP, not depending on a functional respiratory chain. At nM external dTMP the nucleotide is concentrated 100-fold inside the mt matrix. Export of thymidine phosphates was much slower and possibly occurred at the level of dTDP. PMID- 17124169 TI - A Legionella pneumophila-translocated substrate that is required for growth within macrophages and protection from host cell death. AB - Legionella pneumophila requires the Dot/Icm protein translocation system to replicate within host cells as a critical component of Legionnaire's pneumonia. None of the known individual substrates of the translocator have been shown to be essential for intracellular replication. We demonstrate here that mutants lacking the Dot/Icm substrate SdhA were severely impaired for intracellular growth within mouse bone marrow macrophages, with the defect absolute in triple mutants lacking sdhA and its two paralogs. The defect caused by the absence of the sdhA family was less severe during growth within Dictyostelium discoideum amoebae, indicating that the requirement for SdhA shows cell-type specificity. Macrophages harboring the L. pneumophila sdhA mutant showed increased nuclear degradation, mitochondrial disruption, membrane permeability, and caspase activation, indicating a role for SdhA in preventing host cell death. Defective intracellular growth of the sdhA(-) mutant could be partially suppressed by the action of caspase inhibitors, but caspase-independent cell death pathways eventually aborted replication of the mutant. PMID- 17124170 TI - A two-step mechanism underlies the planar polarization of regenerating sensory hair cells. AB - The restoration of planar cell polarity is an essential but poorly understood step toward physiological recovery during sensory-organ regeneration. Investigating this issue in the lateral line of the zebrafish, we found that hair cells regenerate in pairs along a single axis established by the restricted localization and oriented division of their progenitors. By analyzing mutants lacking the planar-polarity determinant Vangl2, we ascertained that the uniaxial production of hair cells and the subsequent orientation of their hair bundles are controlled by distinct pathways, whose combination underlies the establishment of hair-cell orientation during development and regeneration. This mechanism may represent a general principle governing the long-term maintenance of planar cell polarity in remodeling epithelia. PMID- 17124171 TI - Direct observation of DNA bending/unbending kinetics in complex with DNA-bending protein IHF. AB - Regulation of gene expression involves formation of specific protein-DNA complexes in which the DNA is often bent or sharply kinked. Kinetics measurements of DNA bending when in complex with the protein are essential for understanding the molecular mechanism that leads to precise recognition of specific DNA-binding sites. Previous kinetics measurements on several DNA-bending proteins used stopped-flow techniques that have limited time resolution of few milliseconds. Here we use a nanosecond laser temperature-jump apparatus to probe, with submillisecond time resolution, the kinetics of bending/unbending of a DNA substrate bound to integration host factor (IHF), an architectural protein from Escherichia coli. The kinetics are monitored with time-resolved FRET, with the DNA substrates end-labeled with a FRET pair. The temperature-jump measurements, in combination with stopped-flow measurements, demonstrate that the binding of IHF to its cognate DNA site involves an intermediate state with straight or, possibly, partially bent DNA. The DNA bending rates range from approximately 2 ms(-1) at approximately 37 degrees C to approximately 40 ms(-1) at approximately 10 degrees C and correspond to an activation energy of approximately 14 +/- 3 kcal/mol. These rates and activation energy are similar to those of a single A:T base pair opening inside duplex DNA. Thus, our results suggest that spontaneous thermal disruption in base-paring, nucleated at an A:T site, may be sufficient to overcome the free energy barrier needed to partially bend/kink DNA before forming a tight complex with IHF. PMID- 17124172 TI - A different function for a member of an ancient and highly conserved cytochrome P450 family: from essential sterols to plant defense. AB - CYP51 sterol demethylases are the only cytochrome P450 enzymes with a conserved function across the animal, fungal, and plant kingdoms (in the synthesis of essential sterols). These highly conserved enzymes, which are important targets for cholesterol-lowering drugs, antifungal agents, and herbicides, are regarded as the most ancient member cytochrome P450 family. Here we present a report of a CYP51 enzyme that has acquired a different function. We show that the plant enzyme AsCYP51H10 is dispensable for synthesis of essential sterols and has been recruited for the production of antimicrobial compounds (avenacins) that confer disease resistance in oats. The AsCyp51H10 gene is synonymous with Sad2, a gene that we previously had defined by mutation as being required for avenacin synthesis. In earlier work, we showed that Sad1, the gene encoding the first committed enzyme in the avenacin pathway (beta-amyrin synthase), had arisen by duplication and divergence of a cycloartenol synthase-like gene. Together these data indicate an intimate evolutionary connection between the sterol and avenacin pathways. Sad1 and Sad2 lie within 70 kb of each other and are expressed specifically in the epidermal cells of the root tip, the site of accumulation of avenacins. These findings raise intriguing questions about the recruitment, coevolution, and regulation of the components of this specialized defense-related metabolic pathway. PMID- 17124173 TI - Relative blindsight in normal observers and the neural correlate of visual consciousness. AB - By using a paradigm based on metacontrast masking, we created experimental conditions in which the subjective report of consciousness differs but the objectively measured ability to discriminate visual targets does not. This approach allowed us to study the neural correlate of consciousness while having performance levels carefully matched in healthy human subjects. A comparison of the neural activity associated with these conditions as measured by functional MRI showed that conscious perception is associated with spatially specific activity in the mid-dorsolateral prefrontal cortex (area 46). Further analysis confirms that this activation is not only free from any performance confound, but is also not driven by differences in the timing of the physical stimuli. Our results suggest that the prefrontal cortex is important for the essentially subjective aspects of conscious perception. PMID- 17124174 TI - Global modulation of chromatin dynamics mediated by dephosphorylation of linker histone H1 is necessary for erythroid differentiation. AB - Differentiation of metazoan cells involves dramatic changes in gene expression patterns and proliferative capacity driven primarily by epigenetic mechanisms. Here we used in vivo photobleaching techniques and biochemical assays to investigate the contribution of alterations in chromatin dynamics to the differentiation of murine erythroleukemia (MEL) cells, a model system for erythroid development. As MEL cells differentiate the binding affinity of all linker histone variants increases, indicative of an overall decrease in chromatin flexibility. Changes in H1(0) binding properties depend on phosphorylation at one or more of three cyclin-dependent kinase sites. The presence of constructs mimicking constitutively phosphorylated H1 results in a significant inhibition in the acquisition of commitment to terminal cell division and the expression of erythroid-specific properties. These data indicate that the progressive loss of cdk activity associated with MEL cell differentiation leads to the accumulation of dephosphorylated linker histones and restricted chromatin flexibility and that these are necessary events in the progression of erythroid differentiation. We present additional data indicating that the presence of phosphorylated H1 has a dominant effect on the binding behavior of other linker histones and propose a model for the role of linker histone phosphorylation in which these modifications act within the context of assembled chromatin. PMID- 17124175 TI - Quantifying the relation between adhesion ligand-receptor bond formation and cell phenotype. AB - One of the fundamental interactions in cell biology is the binding of cell receptors to adhesion ligands, and many aspects of cell behavior are believed to be regulated by the number of these bonds that form. Unfortunately, a lack of methods to quantify bond formation, especially for cells in 3D cultures or tissues, has precluded direct probing of this assumption. We now demonstrate that a FRET technique can be used to quantify the number of bonds formed between cellular receptors and synthetic adhesion oligopeptides coupled to an artificial extracellular matrix. Similar quantitative relations were found between bond number and the proliferation and differentiation of MC3T3-E1 preosteoblasts and C2C12 myoblasts, although the relation was distinct for each cell type. This approach to understanding 3D cell-extracellular matrix interactions will allow one to both predict cell behavior and to use bond number as a fundamental design criteria for synthetic extracellular matrices. PMID- 17124176 TI - Active retrotransposition by a synthetic L1 element in mice. AB - Long interspersed element type 1 (L1) retrotransposons are ubiquitous mammalian mobile elements and potential tools for in vivo mutagenesis; however, native L1 elements are relatively inactive in mice when introduced as transgenes. We have previously described a synthetic L1 element, ORFeus, containing two synonymously recoded ORFs relative to mouse L1. It is significantly more active for retrotransposition in cell culture than all native L1 elements tested. To study its activity in vivo, we developed a transgenic mouse model in which ORFeus expression was controlled by a constitutive heterologous promoter, and we established definitive evidence for ORFeus retrotransposition activity both in germ line and somatic tissues. Germ line retrotransposition frequencies resulting in 0.33 insertions per animal are seen among progeny of ORFeus donor element heterozygotes derived from a single founder, representing a >20-fold increase over native L1 elements. We observe somatic transposition events in 100% of the ORFeus donor-containing animals, and an average of 17 different insertions are easily recovered from each animal; modeling suggests that the number of somatic insertions per animal exceeds this number by perhaps several orders of magnitude. Nearly 200 insertions were precisely mapped, and their distribution in the mouse genome appears random relative to transcription units and guanine-cytosine content. The results suggest that ORFeus may be developed into useful tools for in vivo mutagenesis. PMID- 17124177 TI - NMDA receptor surface mobility depends on NR2A-2B subunits. AB - The NR2 subunit composition of NMDA receptors (NMDARs) varies during development, and this change is important in NMDAR-dependent signaling. In particular, synaptic NMDAR switch from containing mostly NR2B subunit to a mixture of NR2B and NR2A subunits. The pathways by which neurons differentially traffic NR2A- and NR2B-containing NMDARs are poorly understood. Using single-particle and -molecule approaches and specific antibodies directed against NR2A and NR2B extracellular epitopes, we investigated the surface mobility of native NR2A and NR2B subunits at the surface of cultured neurons. The surface mobility of NMDARs depends on the NR2 subunit subtype, with NR2A-containing NMDARs being more stable than NR2B containing ones, and NR2A subunit overexpression stabilizes surface NR2B containing NMDARs. The developmental change in the synaptic surface content of NR2A and NR2B subunits was correlated with a developmental change in the time spent by the subunits within synapses. This suggests that the switch in synaptic NMDAR subtypes depends on the regulation of the receptor surface trafficking. PMID- 17124178 TI - Dendritic compartmentalization of chloride cotransporters underlies directional responses of starburst amacrine cells in retina. AB - The mechanisms in the retina that generate light responses selective for the direction of image motion remain unresolved. Recent evidence indicates that directionally selective light responses occur first in the retina in the dendrites of an interneuron, i.e., the starburst amacrine cell, and that these responses are highly sensitive to the activity of Na-K-2Cl (NKCC) and K-Cl (KCC), two types of chloride cotransporter that determine whether the neurotransmitter GABA depolarizes or hyperpolarizes neurons, respectively. We show here that selective blockade of the NKCC2 and KCC2 cotransporters located on starburst dendrites consistently hyperpolarized and depolarized the starburst cells, respectively, and greatly reduced or eliminated their directionally selective light responses. By mapping NKCC2 and KCC2 antibody staining on these dendrites, we further show that NKCC2 and KCC2 are preferentially located in the proximal and distal dendritic compartments, respectively. Finally, measurements of the GABA reversal potential in different starburst dendritic compartments indicate that the GABA reversal potential at the distal dendrite is more hyperpolarized than at the proximal dendrite due to KCC2 activity. These results thus demonstrate that the differential distribution of NKCC2 on the proximal dendrites and KCC2 on the distal dendrites of starburst cells results in a GABA-evoked depolarization and hyperpolarization at the NKCC2 and KCC2 compartments, respectively, and underlies the directionally selective light responses of the dendrites. The functional compartmentalization of interneuron dendrites may be an important means by which the nervous system encodes complex information at the subcellular level. PMID- 17124179 TI - Disulfide-dependent protein folding is linked to operation of the vitamin K cycle in the endoplasmic reticulum. A protein disulfide isomerase-VKORC1 redox enzyme complex appears to be responsible for vitamin K1 2,3-epoxide reduction. AB - Gamma-carboxylation of vitamin K-dependent proteins is dependent on formation of reduced vitamin K1 (Vit.K1H2) in the endoplasmic reticulum (ER), where it works as an essential cofactor for gamma-carboxylase in post-translational gamma carboxylation of vitamin K-dependent proteins. Vit.K1H2 is produced by the warfarin-sensitive enzyme vitamin K 2,3-epoxide reductase (VKOR) of the vitamin K cycle that has been shown to harbor a thioredoxin-like CXXC center involved in reduction of vitamin K1 2,3-epoxide (Vit.K>O). However, the cellular system providing electrons to the center is unknown. Here data are presented that demonstrate that reduction is linked to dithiol-dependent oxidative folding of proteins in the ER by protein disulfide isomerase (PDI). Oxidative folding of reduced RNase is shown to trigger reduction of Vit.K>O and gamma-carboxylation of the synthetic gamma-carboxylase peptide substrate FLEEL. In liver microsomes, reduced RNase-triggered gamma-carboxylation is inhibited by the PDI inhibitor bacitracin and also by small interfering RNA silencing of PDI in HEK 293 cells. Immunoprecipitation and two-dimensional SDS-PAGE of microsomal membrane proteins demonstrate the existence of a VKOR enzyme complex where PDI and VKORC1 appear to be tightly associated subunits. We propose that the PDI subunit of the complex provides electrons for reduction of the thioredoxin-like CXXC center in VKORC1. We can conclude that the energy required for gamma-carboxylation of proteins is provided by dithiol-dependent oxidative protein folding in the ER and thus is linked to de novo protein synthesis. PMID- 17124180 TI - Molecular mechanisms of transactivation and doxorubicin-mediated repression of survivin gene in cancer cells. AB - Human maintenance DNA cytosine methyltransferase (DNMT1) regulates gene expression in a methylation-dependent and -independent manner. Anti-apoptotic survivin gene down-regulation is mediated by p53 recruitment of DNMT1 to its promoter. Survivin inhibits programmed cell death, regulates cell division, and is expressed in cancer cells. The survivin gene promoter is CG-rich containing several Sp1 canonical, Sp1-like, cell cycle-dependent element/cell cycle gene homology region, and p53-binding sites. Here we demonstrate that Sp1 transcription factor(s) play a role in transcriptional activation of the survivin promoter in Drosophila and human cells. Sp1 inhibition in vivo by mithramycin A leads to down-regulation of a luciferase reporter driven by the human survivin promoter in transfected cells. Mithramycin A or Sp1-specific short interfering RNA down-regulated the endogenous survivin gene expression, confirming Sp1 as the primary determinant for transcriptional activation. Furthermore, immobilized DNMT1 ligand bound to seven consensus amino acids corresponding to the N-terminal region of the Sp class of transcription factors in a phage display analysis. In the co-immunoprecipitation assay, the endogenous Sp1 or Sp3 pulled down DNMT1 and methyltransferase activity. Similarly, a glutathione S-transferase pulldown assay between DNMT1 and Sp1 demonstrates a direct interaction between the two proteins. Fluorescent fusions of DNMT1 and Sp1 co-localized in the mammalian nucleus, thus supporting binary complex formation between both the proteins. The kinetics of survivin promoter occupancy via chromatin immunoprecipitation following doxorubicin treatment show the presence of Sp1 and gradual accumulation of transcriptional repressors p53, DNMT1, histone methyltransferase G9a, and HDAC1 onto the promoter along with histone H3K9me2. These data suggest that the Sp1 transcription factor acts as a platform for recruitment of transcriptional repressors. PMID- 17124181 TI - Quaternary structure constraints on evolutionary sequence divergence. AB - The structurally constrained protein evolution (SCPE) model simulates protein divergence considering protein structure explicitly. The model is based on the observation that protein structure is more conserved during evolution than the sequences encoding for that structure. In the previous work, the SCPE model considered only the tertiary structure. Here we show that the performance of the model is enhanced when the oligomeric structure is taken into account. Our results agree with recent evolutionary studies of oligomeric proteins, which show that conservation of the quaternary structure imposes additional constraints on sequence divergence. The incorporation of protein-protein interactions into protein evolution models may be important in the study of quaternary protein structures and complex protein assemblies. PMID- 17124182 TI - Recurrent gene amplification and soft selective sweeps during evolution of multidrug resistance in malaria parasites. AB - When selection is strong and beneficial alleles have a single origin, local reductions in genetic diversity are expected. However, when beneficial alleles have multiple origins or were segregating in the population prior to a change in selection regime, the impact on genetic diversity may be less clear. We describe an example of such a "soft" selective sweep in the malaria parasite Plasmodium falciparum that involves adaptive genome rearrangements. Amplification in copy number of genome regions containing the pfmdr1 gene on chromosome 5 confer resistance to mefloquine and spread rapidly in the 1990s. Using flanking microsatellite data and real-time polymerase chain reaction determination of copy number, we show that 5-15 independent amplification events have occurred in parasites on the Thailand/Burma border. The amplified genome regions (amplicons) range in size from 14.7 to 49 kb and contain 2-11 genes, with 2-4 copies arranged in tandem. To examine the impact of drug selection on flanking variation, we genotyped 48 microsatellites on chromosome 5 in 326 parasites from a single Thai location. Diversity was reduced in a 170- to 250-kb (10-15 cM) region of chromosomes containing multiple copies of pfmdr1, consistent with hitchhiking resulting from the rapid recent spread of selected chromosomes. However, diversity immediately flanking pfmdr1 is reduced by only 42% on chromosomes bearing multiple amplicons relative to chromosomes carrying a single copy. We highlight 2 features of these results: 1) All amplicon break points occur in monomeric A/T tracts (9-45 bp). Given the abundance of these tracts in P. falciparum, we expect that duplications will occur frequently at multiple genomic locations and have been underestimated as drivers of phenotypic evolution in this pathogen. 2) The signature left by the spread of amplified genome segments is broad, but results in only limited reduction in diversity. If such "soft" sweeps are common in nature, statistical methods based on diversity reduction may be inefficient at detecting evidence for selection in genome-wide marker screens. This may be particularly likely when mutation rate is high, as appears to be the case for gene duplications, and in pathogen populations where effective population sizes are typically very large. PMID- 17124183 TI - HPLC techniques for proteomics analysis--a short overview of latest developments. AB - Due to the complex nature of the proteome, instrumentation and methods development for sample cleanup, fractionation, preconcentration, chromatographic separation and detection becomes urgent for the identification of peptides and proteins. Newly developed techniques and equipment for separation and detection, such as nano-HPLC and multidimensional HPLC for protein and peptide separation, enabled proteomics to experience dynamic growth during the past few years. In any proteomic analysis the most important and sometimes most difficult task is the separation of the complex mixture of proteins or peptides. This review describes some aspects and limitations of HPLC, both multidimensional and one-dimensional, in proteomics research without attempting to discuss all available HPLC methods, which would need far more space than available here. PMID- 17124184 TI - Family emotional climate, depression, emotional triggering of asthma, and disease severity in pediatric asthma: examination of pathways of effect. AB - OBJECTIVES: (a) To assess emotional triggering of pediatric asthma and ascertain its contribution to disease morbidity and functional status; (b) to test whether negative family emotional climate (NFEQ) is associated with depressive and/or anxious symptoms and emotional triggering of asthma attacks in the child. METHOD: Children with asthma (N = 272, 56% male, age 7-17) and their primary caregivers answered together an Asthma Trigger Inventory (Ritz, Steptoe, Bobb, Harris, & Edwards, 2006). Children reported on anxious (STAIC) and depressive (CDI) symptoms and on asthma-related quality of life (PAQLQ). Parent(s) reported on their child's internalizing (CBCL-I) and depressive symptoms (CDI-P). A clinician also rated the child's depression using the structured CDRS-R. Asthma diagnosis was confirmed and disease severity rated according to NHLBI guidelines by an asthma clinician. RESULTS: Path analyses indicated that NFEQ was associated with depressive symptoms, which in turn were associated both directly and indirectly (by way of emotional triggering) with disease severity. Comparison of nested models indicated the possibility of differential roles and pathways for anxious versus depressive symptoms. CONCLUSION: Findings elucidate possible pathways of effect by which family emotional climate and child depressive symptoms may influence pediatric asthma disease severity by way of potentiating emotional triggering of asthma. PMID- 17124185 TI - Blind placements of peripherally inserted antecubital central catheters: initial catheter tip position in relation to carina. AB - BACKGROUND: We investigated how often blind placement of peripherally inserted central catheters (PICCs) through the antecubital veins results in a correct tip location in relation to carina and evaluated the inter-observer agreement in locating the tip of PICCs in plain radiography with digital imaging. METHODS: In this study, 202 suitable chest radiographs with PICCs out of 803 patients were identified. An initial audit on the tip of these catheters in relation to carina was done by a consultant anaesthetist and was recorded as the first observer. The same sets of CXRs were examined by a consultant radiologist and the tips were identified and recorded as the second observer. Inter-observer agreement was assessed. RESULTS: In 75 of 202 (37%), PICCs had a central tip location in relation to the carina. Fifty-five of 131 (42%) right-sided catheters had a central location compared with 20 of 71 (28%) of the left-sided catheters. The tip position for right-sided catheters was most frequently centrally located whereas the tip for left-sided catheters was most commonly positioned in the ipsilateral innominate vein. There was excellent agreement between the observers in reporting the tip of PICCs at all positions (kappa=0.87) including central locations (kappa=0.83). CONCLUSIONS: Right antecubital PICCs are more likely to be placed in the central location in relation to the carina. PICCs inserted through the left antecubital veins need to be pushed further down to aim for a central location. Inter-observer variability in identifying the tip of PICCs is least with the introduction of digital imaging. PMID- 17124186 TI - Successful management of patients with a drug-eluting coronary stent presenting for elective, non-cardiac surgery. AB - This paper describes the management of three patients for elective surgery with drug-eluting stents in the coronary circulation. The risks posed at the time of surgery by such patients include acute coronary syndromes, as a result of stent thrombosis, after cessation of anti-platelet therapy and excessive bleeding from continued anti-platelet therapy. We describe a regime for the management of such patients that successfully avoided these risks in three patients with paclitaxel drug-eluting stents requiring elective non-cardiac surgery. PMID- 17124187 TI - Non-invasive metabolic monitoring of patients under anaesthesia by continuous indirect calorimetry--an in vivo trial of a new method. AB - BACKGROUND: Oxygen uptake is an important form of metabolic monitoring for patients under anaesthesia. In critically ill patients oxygen uptake has been shown to provide valuable clinical information in directed therapy and acts as a useful monitor of cardiovascular dysfunction. A new method of continuous real time monitoring of metabolic gas exchange was tested in patients during anaesthesia. METHODS: Using a standard anaesthetic machine with attached semi closed circle absorber system, oxygen uptake was measured continuously throughout surgery in 30 patients undergoing cardiopulmonary bypass surgery and compared with paired measurements made with the reverse Fick method. The method is an indirect calorimetry technique which uses fresh gas rotameters for control, regulation and measurement of the gas flows into the system, with continuous sampling of mixed exhaust gas. RESULTS: When compared with the reverse Fick method the oxygen uptake showed a mean difference (and sd) of 20.7 ml min(-1) or 12.1% (25.3 ml min(-1)) pre-bypass and 13.9 ml min(-1) or 8.1% (27.0 ml min(-1)) post-bypass. This bias is consistent with previous studies comparing oxygen uptake measured at the mouth against oxygen uptake by reverse Fick, which have shown a difference of approximately 10-15% accounted for by the consumption of oxygen by lung tissue. CONCLUSIONS: As the method allows continuous measurement of gas exchange and can be adapted to a modern anaesthetic workstation it is an attractive method for use in clinical setting. PMID- 17124188 TI - Blood-brain barrier leakage may lead to progression of temporal lobe epilepsy. AB - Leakage of the blood-brain barrier (BBB) is associated with various neurological disorders, including temporal lobe epilepsy (TLE). However, it is not known whether alterations of the BBB occur during epileptogenesis and whether this can affect progression of epilepsy. We used both human and rat epileptic brain tissue and determined BBB permeability using various tracers and albumin immunocytochemistry. In addition, we studied the possible consequences of BBB opening in the rat for the subsequent progression of TLE. Albumin extravasation in human was prominent after status epilepticus (SE) in astrocytes and neurons, and also in hippocampus of TLE patients. Similarly, albumin and tracers were found in microglia, astrocytes and neurons of the rat. The BBB was permeable in rat limbic brain regions shortly after SE, but also in the latent and chronic epileptic phase. BBB permeability was positively correlated to seizure frequency in chronic epileptic rats. Artificial opening of the BBB by mannitol in the chronic epileptic phase induced a persistent increase in the number of seizures in the majority of rats. These findings indicate that BBB leakage occurs during epileptogenesis and the chronic epileptic phase and suggest that this can contribute to the progression of epilepsy. PMID- 17124189 TI - Surgical treatment of drug-resistant nocturnal frontal lobe epilepsy. AB - Of the cases with nocturnal frontal lobe epilepsy (NFLE) approximately 30% are refractory to antiepileptic medication, with several patients suffering from the effects of both ongoing seizures and disrupted sleep. From a consecutive series of 522 patients operated on for drug-resistant focal epilepsy, 21 cases (4%), whose frontal lobe seizures occurred almost exclusively (>90%) during sleep, were selected. All patients underwent a comprehensive pre-surgical evaluation, which included history, interictal EEG, scalp video-EEG monitoring, high-resolution MRI and, when indicated, invasive recording by stereo-EEG (SEEG). There were 11 males and 10 females, whose mean age at seizure onset was 6.2 years, mean age at surgery was 24.7 years and seizure frequency ranged from <20/month to >300/month. Nine patients reported excessive daytime sleepiness (EDS). Prevalent ictal clinical signs were represented by asymmetric posturing (6 cases), hyperkinetic automatisms (10 cases), combined tonic posturing and hyperkinetic automatisms (4 cases) and mimetic automatisms (1 case). All patients reported some kind of subjective manifestations. Interictal and ictal EEG provided lateralizing or localizing information in most patients. MRI was unrevealing in 10 cases and it showed a focal anatomical abnormality in one frontal lobe in 11 cases. Eighteen patients underwent a SEEG evaluation to better define the epileptogenic zone (EZ). All patients received a microsurgical resection in one frontal lobe, tailored according to pre-surgical evaluations. Two patients were operated on twice owing to poor results after the first resection. Histology demonstrated a Taylor-type focal cortical dysplasia (FCD) in 16 patients and an architectural FCD in 4. In one case no histological change was found. After a post-operative follow-up of at least 12 months (mean 42.5 months) all the 16 patients with a Taylor's FCD were in Engel's Class Ia and the other 5 patients were in Engel's Classes II or III. After 6 months post-surgery EDS had disappeared in the 9 patients who presented this complaint pre-operatively. It is concluded that patients with drug-resistant, disabling sleep-related seizures of frontal lobe origin should be considered for resective surgery, which may provide excellent results both on seizures and on epilepsy-related sleep disturbances. An accurate pre-surgical evaluation, which often requires invasive EEG recording, is mandatory to define the EZ. Further investigation is needed to explain the possible causal relationships between FCD, particularly Taylor-type, and sleep related seizures, as observed in this cohort of NFLE patients. PMID- 17124190 TI - Long-term outcomes in epilepsy surgery: antiepileptic drugs, mortality, cognitive and psychosocial aspects. AB - Assessment of long-term outcomes is essential in brain surgery for epilepsy. Little information exists on long-term non-seizure outcomes after epilepsy surgery. We perform a systematic review and meta-analysis of the evidence on this topic. Our aim was to provide evidence-based estimates of antiepileptic drug, psychosocial, neuropsychological and mortality long-term outcomes following epilepsy surgery, and to identify sources of variation in published results. We searched Medline, Index Medicus, the Cochrane database, bibliographies of reviews, original articles, and book chapters, to identify articles published from 1991 to 2005, containing > or =20 patients of any age, undergoing resective or non-resective epilepsy surgery, and followed for a mean/median of > or =5 years. Two reviewers independently assessed study eligibility and extracted data, resolving disagreements through discussion. Standard meta-analytical techniques were used to pool data. Of the 159 potentially eligible articles reviewed in full text, 35 (22%) fulfilled eligibility criteria; 6 (17%) were controlled studies; 15 (36%) explored antiepileptic drug outcome; 6 (17%) explored mortality; 11 (31%) reported psychosocial outcomes; and 7 (20%) explored neuropsychological outcomes. On an average, 14% [95% confidence interval (CI(95)) = 11-17] of the patients with temporal lobe surgery achieved long-term antiepileptic drug (AED) discontinuation, 50% (CI(95) = 45-55) achieved monotherapy, and 33% remained on polytherapy (CI(95 =) 29-38). In analyses including all types of surgery, on average, 20% (CI(95) = 18-23) achieved long-term AED discontinuation, while 41% (CI(95) = 37-45) were on monotherapy and 31% (CI(95) = 27-35) remained on polytherapy. Children achieved better AED outcomes than adults. Seizure freedom after surgery was associated with lower mortality, but inconsistent mortality outcomes precluded making strong inferences. Non-controlled studies consistently reported improved long-term psychosocial outcomes, but the effect was less clear in controlled studies. Intelligence was unchanged by surgery, but long-term memory outcomes were associated with seizure freedom and side of temporal lobe resection. Few long-term, controlled studies exist. Longer follow-up was associated with lower rates of AED discontinuation, reflecting lower seizure-free rates over time. Cognitive and psychosocial outcomes were similar to those of short-term studies, and the results were influenced by the presence of controls. PMID- 17124191 TI - Mixing pro- and antisaccades in patients with parkinsonian syndromes. AB - Prosaccades and antisaccades were investigated in three groups of patients with parkinsonian syndromes, Parkinson's disease, corticobasal degeneration (CBD) and progressive supranuclear palsy (PSP), and in a control group. Saccade tasks were performed in single-task blocks (i.e. either blocks of prosaccades or blocks of antisaccades) and in mixed-task blocks (i.e. in blocks of randomly interleaved pro- and antisaccades). Saccade latencies and directional errors (misdirected saccades) were analysed in each subject, and we concentrated more specifically on the comparison of error rates in single tasks and in repeated trials of mixed tasks (i.e. mixing costs). The performance of each group in single tasks was largely consistent with previous studies, with normal antisaccade error rates in Parkinson's disease and CBD patients and increased antisaccade error rates in PSP patients. In contrast, a double dissociation was observed in mixed tasks. Parkinson's disease and CBD patients showed a marked increase in prosaccade and antisaccade error rates in repeated trials of mixed tasks, illustrated by increased mixing costs, whereas PSP patients showed similar error rates in single and repeated trials of mixed tasks, i.e. normal mixing costs. These results demonstrate that: (i) antisaccade performances may be differentially affected in mixed tasks and single tasks; (ii) the region of the dorsolateral prefrontal cortex which is crucial for reflexive saccade inhibition does not seem to be involved in the additional processes required in mixed-task conditions; (iii) the study of interleaved pro- and antisaccades may increase the accuracy of the differential diagnosis between these parkinsonian syndromes. PMID- 17124192 TI - Is self-medication with antibiotics in Europe driven by prescribed use? AB - BACKGROUND: Self-medication with antibiotics may increase the risk of inappropriate use and the selection of resistant bacteria. One of the triggers for using self-medication may be past experience with antibiotics prescribed by health professionals. We examined the association between prescribed use and self medication with antibiotics. METHODS: A population survey was conducted in 19 European countries, covering 15,548 respondents. Multinomial logistic regression analysis was used to study the relationship between prescribed use and self medication for all symptoms/diseases and for upper respiratory tract infections (URTIs). RESULTS: The association between prescribed use and self-medication was modified by source of self-medication, region in Europe and education. This association was consistently stronger for self-medication from leftovers than from other sources, primarily directly from a pharmacy. It was stronger also for respondents from Northern/Western Europe than respondents from Eastern Europe and Southern Europe and those with low education. Prescribed use for URTIs (minor ailments such as throat symptom, influenza, etc.) increased the likelihood of self-medication with leftover antibiotics for these symptoms/diseases in all European regions. CONCLUSIONS: Our study shows consistent associations between prescribed use and self-medication with antibiotics from leftovers, but has not been able to support the hypothesis that self-medication from other sources than leftovers is triggered by earlier prescribed use. Preventing leftovers may be one effective way of preventing self-medication. This can be achieved by ensuring that the amount dispensed corresponds to the amount prescribed, by educating patients and by making doctors aware that prescribing for minor ailments may increase the risk of self-medication for such ailments. PMID- 17124193 TI - Pronounced in vitro and in vivo antiretroviral activity of 5-substituted 2,4 diamino-6-[2-(phosphonomethoxy)ethoxy] pyrimidines. AB - OBJECTIVES: To discover new potent and selective anti-human immunodeficiency virus (HIV) acyclic nucleoside phosphonate (ANP) drugs with in vivo antiretroviral activity. METHODS: New acyclic pyrimidine nucleoside phosphonate derivatives that mimic the structure of the anti-HIV purine nucleoside phosphonates 9-(2-phosphonylmethoxyethyl)adenine (PMEA, adefovir) and (R)-9-(2 phosphonylmethoxypropyl)adenine (PMPA, tenofovir) were designed by linking the acyclic side chain of the ANPs through an ether bond to the C-6 position instead of the N-1 position of the pyrimidine ring. The compounds were evaluated against HIV and Moloney murine sarcoma virus (MSV) in cell culture, including a broad variety of HIV-1 clade clinical isolates and relevant mutant (drug-resistant) HIV 1 isolates. Their antiviral activities were correlated and investigated in an in vivo model consisting of MSV-infected newborn mice. MSV-induced tumour formation and associated death were recorded in drug-treated animals. RESULTS: Several 5 substituted 6-[2-(phosphonomethoxy)ethoxy]-2,4-diaminopyrimidine (PMEO-DAPy) analogues were found to inhibit a broad variety of HIV-1 clinical isolates. They showed a more favourable cross-resistance profile to mutant virus isolates than adefovir and tenofovir. There was a close correlation between inhibition of MSV in C3H/3T3 cells and inhibition of HIV-1 in CEM cells. The PMEO-DAPy derivatives potently inhibited MSV-induced tumour cell formation in newborn mice. The 5 methyl analogue PMEO-5-Me-DAPy proved markedly more inhibitory to MSV-induced tumour cell formation and associated animal death than its unsubstituted parent PMEO-DAPy derivative. When compared with adefovir, PMEO-5-Me-DAPy was less toxic and more antivirally active in MSV-infected mice. CONCLUSIONS: PMEO-5-Me-DAPy deserves further (pre)clinical investigations as a candidate anti-HIV drug. PMID- 17124194 TI - Reported versus measured body weight and height of 4-year-old children and the prevalence of overweight. AB - BACKGROUND: In adults, body weight tends to be underestimated when based on self reported data. Whether this discrepancy between measured and reported data exists in healthy young children is unclear. We studied whether parental reported body weight and height of 4-year-old children corresponded with measured body weight and height. In addition, we studied the determinants and the consequences of differences between reported and measured data. METHODS: Data on body weight and height of 864 4-year-old Dutch children born in 1996/1997 enrolled in the Prevention and Incidence of Asthma and Mite Allergy (PIAMA) birth cohort study were collected via a questionnaire and a medical examination. Overweight was defined according to standard international age and gender specific definitions. RESULTS: Mean differences between measured and reported body weight, height, and body mass index (BMI) were small. Parents of children with a low BMI tended to over report body weight while parents of children with a high BMI tended to underreport body weight. Whereas 9.5% of the children were overweight according to reported BMI, the prevalence of overweight was 13.4% based on measured BMI. Over 45% of the overweight children according to measured BMI were missed when reported BMI was used. CONCLUSION: These findings suggest that overweight prevalence rates in children are underestimated when based on reported weight and height. PMID- 17124195 TI - Molecular characterization and oligosaccharide-binding properties of a galectin from the argasid tick Ornithodoros moubata. AB - The argasid tick Ornithodoros moubata is a vector of various viral and borrelian diseases in animals and humans. We report here molecular characterization and oligosaccharide-binding properties of a novel galectin (OmGalec) from this tick. OmGalec consisted of 333 amino acids with a predicted molecular weight of 37.4 kDa. Its amino acid sequence did not contain a signal peptide or transmembrane domain. It possessed tandem-repeated carbohydrate recognition domains, in which the typical motifs important for carbohydrate affinity were conserved. OmGalec was expressed both transcriptionally and translationally at all stages of the tick life cycle and in multiple organs and was abundant in hemocytes, midguts, and reproductive organs, which are of importance in immunity, interaction with pathogens, and development, respectively, suggesting that OmGalec is a multifunctional molecule. The oligosaccharide affinity profile analyzed by applying an automated frontal affinity chromatography system revealed that rOmGalec showed a general feature of the galectin family, i.e. significant affinity for lactosamine-type disaccharides, Galbeta1-3(4)Glc(NAc), via recognition of 4-OH and 6-OH of galactose and 3 (4)-OH of Glc(NAc). Its preference for type I saccharides and alpha1-3GalNAc-containing oligosaccharides might provide clues for identifying its ligands and its potential multiple functions. Our results may contribute to the elucidation of galectin functions in the development and immunity of arthropods and/or vector and pathogen interaction and provide valuable information for the development of novel tick control strategies. PMID- 17124196 TI - Virtual histology and the hunt for the vulnerable plaque. PMID- 17124197 TI - Exercise ventilation inefficiency in heart failure: pathophysiological and clinical significance. AB - Heart failure (HF) is a complex syndrome characterized by myocardial dysfunction and a poor prognosis. Among multiple markers of severity, an exercise ventilation inefficiency has important clinical and prognostic value. The pathophysiology determining exercise ventilatory inefficiency is complex and not definitively clarified. Three different mechanisms have been identified: (i) increased dead space, (ii) early occurrence of lactic acidosis, and (iii) abnormal chemoreflex and/or metaboreflex activity. Besides its prognostic value, abnormal ventilation can be influenced by pharmacological and non-pharmacological therapies such as beta-blockers, selective cyclic 3'-5' guanosine monosphosphate phosphodiesterase inhibitors, physical training, and nocturnal continuous positive airway pressure. There is an increasing interest for the exercise periodic breathing, which is frequently associated with HF syndrome and has prognostic importance. The precise mechanisms sustaining exercise periodic breathing are not fully defined but ventilatory and metabo-haemodynamic hypotheses have been proposed. PMID- 17124198 TI - Uninsured Americans and the new Democratic Congress. PMID- 17124199 TI - Risk assessment after acute coronary syndrome. PMID- 17124200 TI - Educational performance in twins. PMID- 17124201 TI - Opinion leader interventions in social networks. PMID- 17124202 TI - NICE emphasises social care for people with dementia. PMID- 17124203 TI - NICE faces legal challenge over restriction on dementia drugs. PMID- 17124204 TI - More than one antiviral should be stockpiled to fight flu pandemic. PMID- 17124208 TI - Charity highlights decline in health in Gaza. PMID- 17124210 TI - Report on health of migrants to UK shows high risk of TB and HIV. PMID- 17124211 TI - European nations draw up strategy to fight obesity. PMID- 17124215 TI - Incidence of syphilis in UK rises as HIV diagnoses hold steady. PMID- 17124217 TI - EU should strengthen protection against fake drugs, UK report says. PMID- 17124221 TI - Clinically localised prostate cancer. PMID- 17124222 TI - Polyarticular septic arthritis. PMID- 17124223 TI - ABC of obesity. Obesity and cancer. PMID- 17124224 TI - Folic acid, homocysteine, and cardiovascular disease: judging causality in the face of inconclusive trial evidence. PMID- 17124225 TI - How much will Herceptin really cost? PMID- 17124226 TI - Doctors as lapdogs to drug firms: Independence may be most cost effective way to improve health care. PMID- 17124227 TI - Doctors as lapdogs to drug firms: the beast is ourselves. PMID- 17124228 TI - How doctors' anonymity in family courts is under threat: Gornall is way out of date. PMID- 17124229 TI - Osteonecrosis of the jaw and bisphosphonates: editorial was confusing. PMID- 17124230 TI - Knee pain interventions show no net gain. PMID- 17124231 TI - Doctors as lapdogs to drug firms: Not lapdogs, not pit bulls. PMID- 17124232 TI - Statins and outcomes in patients with pneumonia: not only healthy user bias. PMID- 17124233 TI - Predicting mental illness in soldiers: too broad a conclusion. PMID- 17124234 TI - Predicting mental illness in soldiers: pre-deployment screening for vulnerability to post- traumatic stress disorder. PMID- 17124235 TI - Osteonecrosis of the jaw and bisphosphonates: historical lesson from occupational medicine. PMID- 17124236 TI - Doctors leading climate change is self delusion. PMID- 17124237 TI - Statins and outcomes in patients with pneumonia: appreciating bias and precision in study. PMID- 17124238 TI - Statins and outcomes in patients with pneumonia: not the best combination. PMID- 17124239 TI - Influence of three-piece and single-piece designs of two sharp-edge optic hydrophobic acrylic intraocular lenses on the prevention of posterior capsule opacification: a prospective, randomised, long-term clinical trial. AB - BACKGROUND: Posterior capsule opacification (PCO) is still a major long-term complication of modern cataract surgery. We evaluated the impact of sharp-edged intraocular lenses (IOLs) with different haptic designs made from the same hydrophobic acrylic material on posterior and anterior lens capsule opacification. SETTING: Eye clinic of Kaunas University of Medicine, Lithuania. Prospective randomised clinical study. METHODS: Seventy-four eyes of 74 patients scheduled for cataract surgery were included in a prospective randomised clinical study. Thirty-seven eyes of 37 patients received a three-piece acrylic hydrophobic (AcrySof, MA3OBA, Alcon) IOL; and thirty-seven eyes of 37 patients received a one-piece acrylic hydrophobic (AcrySof, SA3OAL, Alcon) IOL. Visual acuity, anterior capsule opacification (ACO), capsular folds, capsulorrhexis/optic overlapping and posterior capsule opacification (PCO) were evaluated. ACO was assessed subjectively. PCO values in the entire IOL optic area and in the central 3 mm optic zone were assessed using a photographic image analysis system (EPCO2000). Follow-ups were performed postoperatively at 1 day, 6 months, 1 year and 2 years. RESULTS: There were no significant differences in best corrected visual acuity, grade of ACO and capsulorrhexis/optic overlapping between IOL types during the follow-up period. Patients in the one-piece acrylic hydrophobic IOL group more frequently presented with capsular folds behind the IOL optic area than those in the three-piece IOL group. In the three-piece acrylic hydrophobic IOL group, PCO values (mean (SD)) of the entire IOL optic area were significantly lower six months postoperative (three-piece: 0.002 (0.009); one-piece: 0.007 (0.017); p=0.04), one year postoperative (three-piece: 0.004 (0.016); one-piece: 0.026 (0.041); p=0.001) as well as one year postoperative in the central 3 mm optic zone (three-piece: 0.000 (0.0002); one piece: 0.019 (0.049); p=0.001). However, two years postoperative, the PCO values of the groups did not show significant differences (entire IOL optic area: three piece, 0.136 (0.223); one-piece, 0.154 (0.190); p=0.18; central zone: three piece, 0.023 (0.065); one-piece: 0.020 (0.039); p=0.44). CONCLUSION: The 2 year follow-up after cataract surgery showed no significant difference in ACO and PCO development between three-piece and one-piece acrylic hydrophobic intraocular lenses. PMID- 17124240 TI - Predictions of postoperative visual outcome in subjects with cataract: a preoperative and postoperative study. AB - AIM: To assess the ability of critical flicker frequency (CFF) and optimal reading speed (ORS) to predict the potential vision in patients with cataract with and without ocular comorbidity. METHODS: The two novel tests were compared with two well established potential vision tests (PVTs), the potential acuity meter (PAM) and the laser interferometer (LI). Measurements were made preoperatively in 1 eye of 88 subjects using the battery of 4 PVTs. Postoperative measurements were made with the CFF and the ORS. The subjects studied were consecutive cases over a 12-month period who fulfilled the inclusion and exclusion criteria, and agreed to participate in this study. RESULTS: CFF was the PVT most resistant to the presence of cataract. Both CFF and ORS give a similar predictive precision in the presence of cataract and ocular comorbidity, although CFF seems more precise when the cataract is dense. CONCLUSIONS: The PAM and the LI showed a limited clinical capability in predicting postoperative visual acuity, particularly with dense opacities. The CFF shows the most promise as a PVT, particularly with dense cataract. Further evaluation is required for both CFF and ORS. PMID- 17124241 TI - Diurnal IOP control with bimatoprost versus latanoprost in exfoliative glaucoma: a crossover, observer-masked, three-centre study. AB - AIM: To evaluate the diurnal intraocular pressure (IOP) control and safety of bimatoprost versus latanoprost in exfoliative glaucoma (XFG). METHODS: One eye of 129 consecutive patients with XFG (mean (SD) age 66.5 (8.3) years) was included in this prospective, observer-masked, three-centre, crossover comparison. After a 4-6 week medicine-free period patients were randomised to bimatoprost or latanoprost monotherapy for 3 months. Patients were then switched to the opposite treatment for another 3 months. At the end of the washout and the treatment periods diurnal IOP was measured at 0800, 1300, and 1800. RESULTS: At baseline the IOP (mean (SD)) was 28.0 (4.0), 26.9 (3.6), and 25.9 (3.6) mm Hg, at the three time points, respectively. Both treatments significantly reduced mean diurnal IOP at month 3. Mean diurnal IOP was 26.9 (3.5) mm Hg at baseline, 17.6 (3.3) mm Hg with bimatoprost, and 18.6 (3.6) mm Hg with latanoprost (p<0.0001). Furthermore, lower IOP values were obtained with bimatoprost at all time points (17.9 (3.4), 17.3 (3.3), and 17.6 (3.5) mm Hg, respectively) compared with latanoprost (18.7 (3.6), 18.5 (3.6), and 18.6 (4.1) mm Hg, respectively). The corresponding mean differences (0.8, 1.1, and 1.0 mm Hg, respectively) were all significant (p<0.001 for each comparison). Significantly more patients with XFG obtained a target diurnal IOP <17 mm Hg with bimatoprost than with latanoprost, 55/123 (45%) v 34/123 (28%); (p = 0.001), and significantly fewer patients were non-responders with bimatoprost than with latanoprost (5 v 13, p = 0.021). More patients reported at least one adverse event with bimatoprost than with latanoprost (58 v 41 at 3 months; p = 0.0003). CONCLUSION: This crossover study suggests that better diurnal IOP control is obtained with bimatoprost than with latanoprost in patients with XFG. PMID- 17124242 TI - A novel method for preserving cultured limbal epithelial cells. AB - AIM: To investigate organ culture preservation of cultured limbal epithelial cells in order to enhance the availability of tissue-engineered epithelia that are used to treat patients with limbal stem cell deficiency. METHODS: Limbal epithelial cells were cultured for 3 weeks on intact amniotic membrane fastened to a polyester membrane carrier. The cultured epithelia were stored for 1 week at 23 degrees C in organ culture medium. The preserved epithelia were then examined using a colorimetric cell viability assay, light microscopy and immunohistochemistry. RESULTS: The viability of the preserved epithelia was 84% (20%), and no statistically significant difference was found compared with non preserved epithelia. In general, the cell borders were maintained, the nuclei showed no sign of degeneration, and the original layered structure was preserved. Mild intercellular oedema was occasionally observed. Expression of p63, K19 and vimentin was maintained. CONCLUSIONS: Cultured limbal epithelial cells can be preserved in organ culture medium for 1 week at room temperature, while maintaining the original layered structure and undifferentiated phenotype. PMID- 17124243 TI - Serious complications of local anaesthesia for cataract surgery: a 1 year national survey in the United Kingdom. AB - BACKGROUND: The techniques of sub-Tenon's, topical and topical-intracameral local anaesthesia (LA) have become common in routine practice. AIMS: This study aimed (i) to estimate the frequency of various LA techniques used in cataract surgery, (ii) to estimate the incidence of severe adverse events associated with each LA technique, and (iii) to document these adverse events. METHODS: This was a prospective, 13 month observational study of routine practice in the UK in 2002 2003. The British Ophthalmological Surveillance Unit sent a monthly mailing to UK ophthalmologists, asking for reports of "potentially sight-threatening or life threatening complications of LA for cataract surgery". Current LA practice was assessed by questionnaire. RESULTS: Cataract surgery comprised 4.1% general anaesthesia, 92.1% LA without sedation and 3.9% LA with sedation. Of the estimated 375 000 LAs 30.6% were peribulbar, 3.5% retrobulbar, 42.6% sub-Tenon's, 1.7% sub-conjunctival, 9.9% topical and 11.0% topical-intracameral LA. "Potentially sight-threatening complications" were mostly associated with retrobulbar and peribulbar techniques and "potentially life-threatening" complications with all techniques except topical/intracameral LA. Eight neurological complications consistent with brainstem anaesthesia were reported: 7 with peribulbar or retrobulbar LA. Poisson regression analysis strongly indicated that rates vary with technique (p<0.001 for "potentially sight-threatening" complications, p = 0.03 for "neurological" complications). Because of likely under-reporting, further complications probably occurred during the survey period. CONCLUSIONS: This large survey found a lower rate of reported serious complications with sub-Tenon's, topical and topical-intracameral LA compared with retrobulbar and peribulbar techniques. These "newer" methods may be preferable for routine cataract surgery. PMID- 17124244 TI - The IOP-lowering effects and mechanism of action of tafluprost in prostanoid receptor-deficient mice. AB - AIM: To clarify the intraocular pressure (IOP)-lowering profile of tafluprost, a newly synthesised prostaglandin F(2alpha) analogue, in mice. METHODS: C57BL/6J, and EP1, EP2, EP3 and postaglandin F (FP) receptor-deficient wild-type (WT), EP1KO, EP2KO, EP3KO and FPKO, respectively mice were bred and acclimatised under a 12-h (6:00-18:00) light-dark cycle. To evaluate effects of tafluprost (0.002%) on IOP at night, a single 3 microl drop of tafluprost solution was applied topically at 18:00 once into one eye in each mouse. IOP was measured 3 h after the application with a microneedle method. To clarify whether endogenous prostaglandin is concerned with the tafluprost-induced IOP reduction, we applied 0.1% diclofenac Na, a cyclo-oxygenase inhibitor or PBS 30 min before the application of tafluprost in WT and EP3KO mice and measured IOP 3 h after the tafluprost application. We also determined whether animals responded predictably to 0.1% bunazosin HCl, a drug known to increase uveoscleral outflow. RESULTS: 3 h after the application of 0.0015% tafluprost, mean (SEM) IOP reductions were 25.8 (2.1)% 26.3 (0.8)% 24.2 (1.4)% 16.5 (1.7)% and -0.9 (1.5)% in WT, EP1KO, EP2KO, EP3KO and FPKO mice, respectively. IOP reductions in EP3KO and FPKO mice were significantly smaller than in WT mice. Pretreatment with diclofenac Na significantly attenuated the IOP lowering effect of tafluprost in WT mice but not in EP3KO mice. Bunazosin HCl lowered IOP significantly in all genotypes by the same amount. CONCLUSION: We conclude that tafluprost lowers IOP through the prostanoid FP receptor. A part of ocular hypotensive effect of tafluprost is attributed to FP receptor-mediated prostaglandin production acting through the prostanoid EP3 receptor. PMID- 17124245 TI - Prevalence and causes of visual impairment and blindness in Sistan-va-Baluchestan Province, Iran: Zahedan Eye Study. AB - AIM: To determine the prevalence and causes of visual impairment and blindness in the Sistan-va-Baluchestan Province of Iran. METHODS: A population-based cross sectional study with a multistage cluster sampling technique was used to identify the study subjects. Visual acuity (VA) was defined for all participants aged >or=10 years. Participants with a VA of <20/60 were examined by an ophthalmologist to determine the causes of low vision or blindness. RESULTS: 5446 (84.0%) of the invited people were examined. The prevalence of visual impairment (VA <20/60) was 6.81% (95% CI 5.91% to 7.71%) and of bilateral blindness (VA <3/60) was 0.79% (95% CI 0.50% to 1.08%). Visual impairment increased with age and illiteracy. Bilateral blindness doubled in women aged >40 years. The causes of visual impairment and blindness were cataract (37.7%), corneal opacity (15.0%), amblyopia (15.0%), glaucoma (5.7%) and hyperopia (5.0%). 39.5% of the visual impairment cases were potentially curable. CONCLUSION: The estimated magnitude of visual impairment and blindness was much higher than our expectations. Further investigation of the pattern of vision loss in women and children, particularly as a result of trachoma and amblyopia, is warranted. Implementation of measures to treat curable cases of the study population can improve the situation in the region dramatically. PMID- 17124246 TI - Proteomics analysis and protein expression during sporozoite excystation of Cryptosporidium parvum (Coccidia, Apicomplexa). AB - Cryptosporidiosis, caused by coccidian parasites of the genus Cryptosporidium, is a major cause of human gastrointestinal infections and poses a significant health risk especially to immunocompromised patients. Despite intensive efforts for more than 20 years, there is currently no effective drug treatment against these protozoa. This study examined the zoonotic species Cryptosporidium parvum at two important stages of its life cycle: the non-excysted (transmissive) and excysted (infective) forms. To increase our understanding of the molecular basis of sporozoite excystation, LC-MS/MS coupling with a stable isotope N-terminal labeling strategy using iTRAQ reagents was used on soluble fractions of both non excysted and excysted sporozoites, i.e. sporozoites both inside and outside oocysts were examined. Sporozoites are the infective stage that penetrates small intestinal enterocytes. Also to increase our knowledge of the C. parvum proteome, shotgun sequencing was performed on insoluble fractions from both non-excysted and excysted sporozoites. In total 303 C. parvum proteins were identified, 56 of which, hitherto described as being only hypothetical proteins, are expressed in both excysted and non-excysted sporozoites. Importantly we demonstrated that the expression of 26 proteins increases significantly during excystation. These excystation-induced proteins included ribosomal proteins, metabolic enzymes, and heat shock proteins. Interestingly three Apicomplexa-specific proteins and five Cryptosporidium-specific proteins augmented in excysted invasive sporozoites. These eight proteins represent promising targets for developing vaccines or chemotherapies that could block parasite entry into host cells. PMID- 17124247 TI - Identification of the ligands of protein interaction domains through a functional approach. AB - The identification of protein-protein interaction networks has often given important information about the functions of specific proteins and on the cross talk among metabolic and regulatory pathways. The availability of entire genome sequences has rendered feasible the systematic screening of collections of proteins, often of unknown function, aimed to find the cognate ligands. Once identified by genetic and/or biochemical approaches, the interaction between two proteins should be validated in the physiologic environment. Herein we describe an experimental strategy to screen collections of protein-protein interaction domains to find and validate candidate interactors. The approach is based on the assumption that the overexpression in cultured cells of protein-protein interaction domains, isolated from the context of the whole protein, could titrate the endogenous ligand and, in turn, exert a dominant negative effect. The identification of the ligand could provide us with a tool to check the relevance of the interaction because the contemporary overexpression of the isolated domain and of its ligand could rescue the dominant negative phenotype. We explored this approach by analyzing the possible dominant negative effects on the cell cycle progression of a collection of phosphotyrosine binding (PTB) domains of human proteins. Of 47 PTB domains, we found that the overexpression of 10 of them significantly interfered with the cell cycle progression of NIH3T3 cells. Four of them were used as baits to identify the cognate interactors. Among these proteins, CARM1, interacting with the PTB domain of RabGAP1, and EF1alpha, interacting with RGS12, were able to rescue the block of the cell cycle induced by the isolated PTB domain of the partner protein, thus confirming in vivo the relevance of the interaction. These results suggest that the described approach can be used for the systematic screening of the ligands of various protein protein interaction domains also by using different biological assays. PMID- 17124248 TI - Adalimumab: a new modality for Behcet's disease? PMID- 17124249 TI - Is shared care with annual hospital review better value for money than predominantly hospital-based care in patients with established stable rheumatoid arthritis? AB - OBJECTIVE: To assess the cost effectiveness and cost effectiveness acceptability of symptom control delivered by shared care (SCSC) and aggressive treatment delivered in hospital (ATH) for established rheumatoid arthritis (RA). METHODS: Economic data were collected within the British Rheumatoid Outcome Study Group randomised controlled trial of SCSC and ATH. A broad perspective was used (UK National Health Service, social support services and patients). Cost per quality adjusted life year (QALY) gained, net benefit statistics and cost effectiveness acceptability curves were estimated. Costs and outcomes were discounted at 3.5%. Sensitivity analysis tested the robustness of the results to analytical assumptions. RESULTS: The mean (SD) cost per person was 4540 pounds (4700) in the SCSC group and 4440 pounds (4900) in the ATH group. The mean (SD) QALYs per person for 3 years were 1.67 (0.56) in the SCSC group and 1.60 (0.60) in the ATH group. If decision makers are prepared to pay > or = 2000 pounds to gain 1 QALY, SCSC is likely to be cost effective in 60-90% of cases. CONCLUSIONS: The primary economic analysis and sensitivity analyses indicate that SCSC is likely to be more cost effective than ATH in 60-90% of cases. This result seems to be robust to assumptions required by the analysis. This study is one of a limited number of randomised controlled trials to collect detailed resource use and health status data and estimate the costs and QALYs of treatment for established RA. This trial is one of the largest RA studies to use the EuroQol. PMID- 17124250 TI - Attachment to laminin-111 facilitates transforming growth factor beta-induced expression of matrix metalloproteinase-3 in synovial fibroblasts. AB - BACKGROUND: In the synovial membrane of patients with rheumatoid arthritis (RA), a strong expression of laminins and matrix degrading proteases was reported. AIM: To investigate the regulation of matrix metalloproteinases (MMPs) in synovial fibroblasts (SFs) of patients with osteoarthritis (OA) and RA by attachment to laminin-1 (LM-111) and in the presence or absence of costimulatory signals provided by transforming growth factor beta (TGFbeta). METHODS: SFs were seeded in laminin-coated flasks and activated by addition of TGFbeta. The expression of genes was investigated by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR), immunocytochemistry and ELISA, and intracellular signalling pathways by immunoblotting, and by poisoning p38MAPK by SB203580, MEK-ERK by PD98059 and SMAD2 by A-83-01. RESULTS: Attachment of SF to LM-111 did not activate the expression of MMPs, but addition of TGFbeta induced a fivefold higher expression of MMP-3. Incubation of SF on LM-111 in the presence of TGFbeta induced a significant 12-fold higher expression of MMP-3 mRNA, and secretion of MMP-3 was elevated 20-fold above controls. Functional blocking of LM-111-integrin interaction reduced the laminin-activated MMP-3 expression significantly. Stimulation of SF by LM-111 and TGFbeta activated the p38MAPK, ERK and SMAD2 pathways, and inhibition of these pathways by using SB203580, PD98059 or A-83-01 confirmed the involvement of these pathways in the regulation of MMP-3. CONCLUSION: Attachment of SF to LM-111 by itself has only minor effects on the expression of MMP-1 or MMP-3, but it facilitates the TGFbeta-induced expression of MMP-3 significantly. This mode of MMP-3 induction may therefore contribute to inflammatory joint destruction in RA independent of the proinflammatory cytokines interleukin (IL)1beta or tumour necrosis factor (TNF)alpha. PMID- 17124251 TI - On the reactivity of monoclonal antibodies specific for different forms of HLA class I molecules. PMID- 17124252 TI - Microchimeric cells: guardians or actors of immunity in scleroderma? PMID- 17124253 TI - The risk of dementia with increased body mass index. AB - BACKGROUND: identification of modifiable risk factors is crucial in the prevention of dementia, given its limited treatment options. Studies on increased body mass index (BMI) as a risk factor for dementia show conflicting results. METHODS: we systematically retrieved and reviewed longitudinal population-based studies on increased BMI and dementia using a standard protocol. We searched Medline (1966-2006), Ageline (1978-2006), PsychInfo (1966-2006), CINAHL (1982 2006), and other relevant databases, including the reference lists of the eligible articles for review. Included studies were subjected to a quality assessment protocol. RESULTS: we identified eight studies that met our selection criteria. These studies covered 1,688 cases of dementia from 28,697 participants. After adjustment for age, smoking, comorbidities, and other confounders, four studies presented significantly increased risk of dementia with elevated BMI. CONCLUSION: this systematic review supports the hypothesis that increased BMI is independently associated with increased risk of dementia. Long-term studies to examine the mechanisms underlying the relationship between obesity and dementia are needed. PMID- 17124255 TI - Development and implementation of a nationwide health care quality indicator system in Taiwan. AB - QUALITY ISSUES: Quality is an increasingly important issue to the health care sector. The Taiwanese government also recognizes the need to implement a nationwide health care quality indicator system to strengthen quality surveillance. CHOICE OF SOLUTION: In 1999, the Department of Health funded a 2 year project led by the Taiwan Healthcare Executive College to develop a comprehensive performance assessment system, subsequently named as Taiwan Healthcare Indicator Series (THIS). The series includes four categories of indicators, namely outpatient, in-patient, emergency care, and intensive care, and has 139 items in total. IMPLEMENTATION: The system was officially launched in 2001. Participation is voluntary. The Taiwan Healthcare Executive College processes the data and provides feedback to the participating hospitals. The information is for the participating hospitals' own use and is not released to the public. EVALUATION: Participating hospitals have increased from 45 in 2001 to 227 in 2006 and now constitute approximately 50% of the total hospital population in Taiwan. The reporting rate averaged 77.7% in 2004. The first five most reported indicators are the percentage of first-visit outpatients to outpatient clinics, the average length of in-patient stay, the nosocomial infection rate, the occupancy rate, and the crude mortality rate. LESSONS LEARNED: How the data are interpreted and how data interpretation can lead to quality improvement are the principal concerns of participating hospitals. In light of the success of the indicator series, the Bureau of National Health Insurance (BNHI) of Taiwan has proposed participation in the series as being one of the criteria to be reimbursed for quality. PMID- 17124256 TI - Domestic recycling of kitchen wastes: an additional health hazard for householders? PMID- 17124257 TI - Trends in the prescription and cost of diabetic medications and monitoring equipment in England 1991-2004. AB - BACKGROUND: To report the trend in prescriptions and cost of antidiabetic drugs and glucose monitoring equipment in England from 1991 to 2004. METHODS: We analysed data on all community antidiabetic drug prescriptions in England collated from the Prescription Cost Analysis system. RESULTS: The total number of diabetes prescriptions (medicines and monitoring) rose from 7,613,000 (1991) to 24,325,640 (2004) (>300% increase). Meanwhile, total costs increased by 650%. Insulins are the biggest contributor to cost followed by monitoring equipment and then oral medications. Three times as many items of oral tablets are prescribed than insulins. Metformin accounts for 40% of all diabetic drug dispensations but only 7% of the costs. More is spent on glitazones now than on either metformin or sulphonylureas. CONCLUSIONS: There has been a substantial increase in the cost of managing diabetes in the community. Costs are likely to continue to rise in the future, as the prevalence of diabetes increases and through more aggressive identification and management of patients with diabetes in the hope of reducing the even more costly complications. The cost implications of glucose monitoring merits further study. PMID- 17124258 TI - Life 20 years after unsuccessful infertility treatment. AB - BACKGROUND: This study explores the long-term experience of involuntary childlessness among 14 Swedish women 20 years after their infertility treatment. METHODS: In-depth interviews were conducted. RESULTS: The childlessness had had a strong impact on all the women's lives and was for all a major life theme. The effects were experienced both on a personal level and on interpersonal and social levels. Half of the women were separated, and in all but one, sexual life was affected in negative and long-lasting ways. The effects of childlessness were especially increased at the time the study was conducted, as the women's peer group was entering the 'grandparent phase'. Many coped with their childlessness by caring for others, such as the children of friends or relatives, elderly parents or animals. CONCLUSION: These findings represent a small sample, but they point towards the need for developing models of counselling and support that stimulate self-reflection and strengthen personal resources and empowerment for individuals and couples experiencing involuntary childlessness. PMID- 17124259 TI - Appropriate remedial action? medical students, medical schools, and smoking and health education in New York and the United States, 1964-87. AB - The Surgeon General's 1964 report on smoking and health, which declared that cigarette smoking was a cause of lung cancer, is considered a landmark in the history of medicine and public health. This article examines the impact of the report on medical student education by reviewing how the relationship between smoking and lung cancer was presented in medical school textbooks and syllabi between 1964 and 1987, changes in hospital smoking regulations and doctors' attitudes toward smoking following the publication of the report, and medical students' smoking patterns and attitudes toward cigarette smoking in the years after 1964. Although it provided some advanced students with additional insight into mechanisms of pathogenesis related to smoking, the education that many medical students received seems to have been neither a primary influence on their smoking patterns nor an important source of their scientific understanding of the causal link between smoking and lung cancer for at least a decade following the publication of the Surgeon General's report. PMID- 17124260 TI - Can all residual force enhancement be explained by sarcomere non-uniformities? PMID- 17124262 TI - Carbonic anhydrase inhibition prevents and reverts cardiomyocyte hypertrophy. AB - Hypertrophic cardiomyocyte growth contributes substantially to the progression of heart failure. Activation of the plasma membrane Na+-H+ exchanger (NHE1) and Cl- HCO3- exchanger (AE3) has emerged as a central point in the hypertrophic cascade. Both NHE1 and AE3 bind carbonic anhydrase (CA), which activates their transport flux, by providing H+ and HCO3-, their respective transport substrates. We examined the contribution of CA activity to the hypertrophic response of cultured neonatal and adult rodent cardiomyocytes. Phenylephrine (PE) increased cell size by 37 +/- 2% and increased expression of the hypertrophic marker, atrial natriuretic factor mRNA, twofold in cultured neonatal rat cardiomyocytes. Cell size was also increased in adult cardiomyocytes subjected to angiotensin II or PE treatment. These effects were associated with increased expression of cytosolic CAII protein and the membrane-anchored isoform, CAIV. The membrane-permeant CA inhibitor, 6-ethoxyzolamide (ETZ), both prevented and reversed PE-induced hypertrophy in a concentration-dependent manner in neonate cardiomyocytes (IC50=18 microm). ETZ and the related CA inhibitor methazolamide prevented hypertrophy in adult cardiomyocytes. In addition, ETZ inhibited transport activity of NHE1 and the AE isoform, AE3, with respective EC50 values of 1.2 +/- 0.3 microm and 2.7 +/- 0.3 microm. PE significantly increased neonatal cardiomyocyte Ca2+ transient frequency from 0.33 +/- 0.4 Hz to 0.77 +/- 0.04 Hz following 24 h treatment; these Ca2+ -handling abnormalities were completely prevented by ETZ (0.28 +/- 0.07 Hz). Our study demonstrates a novel role for CA in mediating the hypertrophic response of cardiac myocytes to PE and suggests that CA inhibition represents an effective therapeutic approach towards mitigation of the hypertrophic phenotype. PMID- 17124263 TI - Interactions between pairs of transcranial magnetic stimuli over the human left dorsal premotor cortex differ from those seen in primary motor cortex. AB - A single TMS pulse (110% resting motor threshold, RMT) to the left dorsal premotor cortex (PMd) (CS2) suppresses the amplitude of motor evoked potentials (MEPs) from a test pulse (TS) over the right motor cortex (M1), and facilitates MEPs from the left motor cortex. We probed how this interaction was changed by a prior conditioning pulse over PMd (CS1) using a paired pulse TMS design. In the main experiments, the intensity of CS1 was 80% RMT. Basal suppression of right M1 was removed when CS1-CS2 was 1 ms or 5 ms with a similar tendency at 15 ms. Basal facilitation of left M1 was suppressed at CS1-CS2 of 5 ms. A similar time course was seen if CS2 was increased to 100% RMT, but there was no significant effect if CS1 was 70% RMT. Preconditioning PMd with continuous or intermittent theta burst repetitive TMS (cTBS, iTBS) abolished the basal CS2-TS interaction between premotor and motor cortices. Finally, if very short interstimulus intervals between CS1 and CS2 were explored to detect interactions similar to I-wave facilitation in M1, we found that the basal suppression of right M1 was abolished at CS1-CS2 intervals of 1.8 and 2.8 ms. We suggest that paired pulse TMS may be capable of investigating properties of intrinsic circuits in PMd and that their properties differ from those in the nearby M1. Paired TMS may be a useful method of studying the excitability of intrinsic circuits in non-primary areas of the motor system. PMID- 17124264 TI - Pinch-current injection defines two discharge profiles in mouse superficial dorsal horn neurones, in vitro. AB - Neurones in the superficial dorsal horn (SDH) are a major target for nociceptive afferents and play an important role in pain processing. One approach to understanding the role of SDH neurones has been to study their action potential (AP) discharge in spinal cord slices during injection of depolarizing step currents. Four or five neurone subpopulations are typically identified based on AP discharge, with various roles proposed for each in pain processing. During noxious peripheral stimulation in vivo, however, SDH neurones are activated via synaptic inputs. This produces a conductance change with different somato dendritic distributions and temporal characteristics to that provided by a somatic step-current injection. Here we introduce an alternative approach to studying SDH neurone discharge under in vitro conditions. We recorded voltage clamp responses in SDH neurones, in vivo, during noxious mechanical stimulation of the hindpaw (1 s pinch, approximately 100 g mm(-2)). From these recordings a representative 'pinch-current' was selected and subsequently injected into SDH neurones in spinal cord slices (recording temperature 32 degrees C). Pinch current-evoked discharge was compared to that evoked by rectangular step-current injections. Pinch- and step-current-evoked AP discharge frequency was highly correlated (r2 = 0.61). This was also true for rheobase current comparisons (r2 = 0.61). Conversely, latency to discharge and discharge duration were not correlated when step- and pinch-current responses were compared. When neurones were grouped according to step-current-evoked discharge, five distinct patterns were apparent (tonic firing, initial bursting, delayed firing, single spiking, and reluctant firing). In contrast, pinch-current responses separated into two clear patterns of activity (robust and resistant firing). During pinch-current injection, tonic-firing and initial-bursting neurones exhibited robust AP discharge with similar characteristics. In contrast, single-spiking and reluctant firing neurones were resistant to AP discharge. Delayed-firing neurones exhibited pinch-current responses that were transitional between those of tonic firing/initial-bursting and single-spiking/reluctant-firing neurones. Injection of digitally filtered pinch-currents indicated that transient current fluctuations are necessary for robust repetitive discharge in initial-bursting neurones. These data suggest the functional significance of the diverse step current-evoked firing patterns, previously reported in SDH neurones remains to be fully understood. When a 'facsimile' current profile or pinch-current is used in place of step-currents, AP discharge diversity is much reduced. PMID- 17124265 TI - Response variability of marmoset parvocellular neurons. AB - This study concerns the properties of neurons carrying signals for colour vision in primates. We investigated the variability of responses of individual parvocellular lateral geniculate neurons of dichromatic and trichromatic marmosets to drifting sinusoidal luminance and chromatic gratings. Response variability was quantified by the cycle-to-cycle variation in Fourier components of the response. Averaged across the population, the variability at low contrasts was greater than predicted by a Poisson process, and at high contrasts the responses were approximately 40% more variable than responses at low contrasts. The contrast-dependent increase in variability was nevertheless below that expected from the increase in firing rate. Variability falls below the Poisson prediction at high contrast, and intrinsic variability of the spike train decreases as contrast increases. Thus, while deeply modulated responses in parvocellular cells have a larger absolute variability than weakly modulated ones, they have a more favourable signal: noise ratio than predicted by a Poisson process. Similar results were obtained from a small sample of magnocellular and koniocellular ('blue-on') neurons. For parvocellular neurons with pronounced colour opponency, chromatic responses were, on average, less variable (10-15%, p<0.01) than luminance responses of equal magnitude. Conversely, non-opponent parvocellular neurons showed the opposite tendency. This is consistent with a supra-additive noise source prior to combination of cone signals. In summary, though variability of parvocellular neurons is largely independent of the way in which they combine cone signals, the noise characteristics of retinal circuitry may augment specialization of parvocellular neurons to signal luminance or chromatic contrast. PMID- 17124266 TI - Enhanced macroscopic desensitization shapes the response of alpha4 subtype containing GABAA receptors to synaptic and extrasynaptic GABA. AB - Up-regulation of the GABAA receptor alpha4 subunit subtype has been consistently shown in multiple animal models of chronic epilepsy. This isoform is expressed in both thalamus and hippocampus and is likely to play a significant role in regulating corticothalamic and hippocampal rhythms. However, little is known about its physiological properties, thus limiting understanding of the role of alpha4 subtype-containing GABAA receptors in normal and abnormal physiology. We used rapid GABA application to recombinant GABAA receptors expressed in HEK293T cells to compare the macroscopic kinetic properties of alpha4beta3gamma2L receptors to those of the more widely distributed alpha1beta3gamma2L receptors. These receptor currents had similar peak current amplitudes and GABA EC50 values. However, alpha4beta3gamma2L currents activated more slowly when exposed to submaximal GABA concentrations, had more fast desensitization (tau = 15-100 ms), and had less residual current during long GABA applications. In addition, alpha4beta3gamma2L currents deactivated more slowly than alpha1beta3gamma2L currents. Peak currents evoked by repetitive, brief GABA applications were more strongly attenuated for alpha4beta3gamma2L currents than alpha1beta3gamma2L currents. Moreover, the time required to recover from desensitization was prolonged in alpha4beta3gamma2L currents compared to alpha1beta3gamma2L currents. We also found that exposure to prolonged low levels of GABA, similar to those that might be present in the extrasynaptic space, greatly suppressed the response of alpha4beta3gamma2L currents to higher concentrations of GABA, while alpha1beta3gamma2L currents were less affected by exposure to low levels of GABA. Taken together, these data suggest that alpha4beta3gamma2L receptors have unique kinetic properties that limit the range of GABA applications to which they can respond maximally. While similar to alpha1beta3gamma2L receptors in their ability to respond to brief and low frequency synaptic inputs, alpha4beta3gamma2L receptors are less efficacious when exposed to prolonged tonic GABA or during repetitive stimulation, as may occur during learning and seizures. PMID- 17124267 TI - Maturation of rat cerebellar Purkinje cells reveals an atypical Ca2+ channel current that is inhibited by omega-agatoxin IVA and the dihydropyridine (-)-(S) Bay K8644. AB - To determine if the properties of Ca2+ channels in cerebellar Purkinje cells change during postnatal development, we recorded Ca2+ channel currents from Purkinje cells in cerebellar slices of mature (postnatal days (P) 40-50) and immature (P13-20) rats. We found that at P40-50, the somatic Ca2+ channel current was inhibited by omega-agatoxin IVA at concentrations selective for P-type Ca2+ channels (approximately 85%; IC50, <1 nM) and by the dihydropyridine (-)-(S)-Bay K8644 (approximately 70%; IC50, approximately 40 nM). (-)-(S)-Bay K8644 is known to activate L-type Ca2+ channels, but the decrease in current was not secondary to the activation of L-type channels because inhibition by (-)-(S)-Bay K8644 persisted in the presence of the L-type channel blocker (R,S)-nimodipine. By contrast, at P13-20, the current was inhibited by omega-agatoxin IVA (approximately 86%; IC50, approximately 1 nM) and a minor component was inhibited by (R,S)-nimodipine (approximately 8%). The dihydropyridine (-)-(S)-Bay K8644 had no clear effect when applied alone, but in the presence of (R,S)-nimodipine it reduced the current (approximately 40%), suggesting that activation of L-type channels by (-)-(S)-Bay K8644 masks its inhibition of non-L-type channels. Our findings indicate that Purkinje neurons express a previously unrecognized type of Ca2+ channel that is inhibited by omega-agatoxin IVA, like prototypical P-type channels, and by (-)-(S)-Bay K8644, unlike classical P-type or L-type channels. During maturation, there is a decrease in the size of the L-type current and an increase in the size of the atypical Ca2+ channel current. These changes may contribute to the maturation of the electrical properties of Purkinje cells. PMID- 17124268 TI - Neurotransmitter mechanisms mediating low-glucose signalling in cocultures and fresh tissue slices of rat carotid body. AB - The mammalian carotid body (CB) is a polymodal chemosensor which can detect low blood glucose (hypoglycaemia), leading to increased afferent discharge and activation of counter-regulatory autonomic pathways. The underlying neurotransmitter mechanisms are unknown and controversy surrounds whether the action of low glucose is direct or indirect. To address this, we used a coculture model containing functional chemosensory units of rat CB receptor (type I) cell clusters and afferent petrosal neurones (PN). During perforated-patch, whole-cell recordings, low glucose (0-2 mM) stimulated sensory discharge in cocultured PN. When the background P(O2) was lowered to levels typical of arterial blood (approximately 90 mmHg), robust PN chemoexcitation could be induced by physiological hypoglycaemia (3.3-4 mM glucose). These sensory responses were reversibly inhibited by a combination of purinergic (suramin, 50 microM) and nicotinic (mecamylamine, 1 microM) receptor blockers, suggesting that transmission depended on corelease of ATP and ACh. Hypoglycaemic responses were additive with those evoked by hypoxia or hypercapnia; further, they could be potentiated by the GABAB receptor blocker (CGP 55845) and inhibited by 5-HT2A receptor blockers (ketanserin or ritanserin). During paired simultaneous recordings from a PN and a type I cell in an adjacent cluster, the afferent PN response coincided with type I cell depolarization, which was associated with a decrease in input resistance. In fresh tissue slices of rat CB, low glucose stimulated ATP secretion as determined by the luciferin-luciferase assay; this secretion was cadmium sensitive, potentiated by CGP 55845, and inhibited by ketanserin. Taken together these data indicate that CB receptors act as direct glucosensors, and that processing of hypoglycaemia utilizes similar neurotransmitter and neuromodulatory mechanisms as hypoxia. PMID- 17124269 TI - Intrauterine growth restriction delays cardiomyocyte maturation and alters coronary artery function in the fetal sheep. AB - There is now extensive evidence suggesting that intrauterine perturbations are linked with an increased risk of developing cardiovascular disease. Human epidemiological studies, supported by animal models, have demonstrated an association between low birth weight, a marker of intrauterine growth restriction (IUGR), and adult cardiovascular disease. However, little is known of the early influence of IUGR on the fetal heart and vessels. The aim of this study was to determine the effects of late gestational IUGR on coronary artery function and cardiomyocyte maturation in the fetus. IUGR was induced by placental embolization in fetal sheep from 110 to 130 days of pregnancy (D110-130); term approximately D147; control fetuses received saline. At necropsy (D130), wire and pressure myography was used to test endothelial and smooth muscle function, and passive mechanical wall properties, respectively, in small branches of left descending coronary arteries. Myocardium was dissociated for histological analysis of cardiomyocytes. At D130, IUGR fetuses (2.7 +/- 0.1 kg) were 28% lighter than controls (3.7 +/- 0.3 kg; P = 0.02). Coronary arteries from IUGR fetuses had enhanced responsiveness to the vasoconstrictors, angiotensin II and the thromboxane analogue U46619, than controls (P < 0.01). Endothelium-dependent and independent relaxations were not different between groups. Coronary arteries of IUGR fetuses were more compliant (P = 0.02) than those of controls. The incidence of cardiomyocyte binucleation was lower in the left ventricles of IUGR fetuses (P = 0.02), suggestive of retarded cardiomyocyte maturation. We conclude that late gestational IUGR alters the reactivity and mechanical wall properties of coronary arteries and cardiomyocyte maturation in fetal sheep, which could have lifelong implications for cardiovascular function. PMID- 17124270 TI - Protease-activated receptor 2 sensitizes the transient receptor potential vanilloid 4 ion channel to cause mechanical hyperalgesia in mice. AB - Exacerbated sensitivity to mechanical stimuli that are normally innocuous or mildly painful (mechanical allodynia and hyperalgesia) occurs during inflammation and underlies painful diseases. Proteases that are generated during inflammation and disease cleave protease-activated receptor 2 (PAR2) on afferent nerves to cause mechanical hyperalgesia in the skin and intestine by unknown mechanisms. We hypothesized that PAR2-mediated mechanical hyperalgesia requires sensitization of the ion channel transient receptor potential vanilloid 4 (TRPV4). Immunoreactive TRPV4 was coexpressed by rat dorsal root ganglia (DRG) neurons with PAR2, substance P (SP) and calcitonin gene-related peptide (CGRP), mediators of pain transmission. In PAR2-expressing cell lines that either naturally expressed TRPV4 (bronchial epithelial cells) or that were transfected to express TRPV4 (HEK cells), pretreatment with a PAR2 agonist enhanced Ca2+ and current responses to the TRPV4 agonists phorbol ester 4alpha-phorbol 12,13-didecanoate (4alphaPDD) and hypotonic solutions. PAR2-agonist similarly sensitized TRPV4 Ca2+ signals and currents in DRG neurons. Antagonists of phospholipase Cbeta and protein kinases A, C and D inhibited PAR2-induced sensitization of TRPV4 Ca2+ signals and currents. 4alphaPDD and hypotonic solutions stimulated SP and CGRP release from dorsal horn of rat spinal cord, and pretreatment with PAR2 agonist sensitized TRPV4-dependent peptide release. Intraplantar injection of PAR2 agonist caused mechanical hyperalgesia in mice and sensitized pain responses to the TRPV4 agonists 4alphaPDD and hypotonic solutions. Deletion of TRPV4 prevented PAR2 agonist-induced mechanical hyperalgesia and sensitization. This novel mechanism, by which PAR2 activates a second messenger to sensitize TRPV4-dependent release of nociceptive peptides and induce mechanical hyperalgesia, may underlie inflammatory hyperalgesia in diseases where proteases are activated and released. PMID- 17124271 TI - Simultaneous measurement of current and calcium in the ultraviolet-sensitive cones of zebrafish. AB - In rods and visible cone photoreceptors, multiple measurements cannot be made of intracellular Ca2+ concentration from the same cell using fluorescent dyes, because a single exposure of the measuring light bleaches too large a fraction of the rod or cone photopigment. We have therefore identified and characterized UV sensitive cones of the zebrafish, whose wavelength of maximum sensitivity is at 360 nm which is far enough from the wavelength of our measuring light (514.5 nm) so that it has been possible to make multiple determinations of photocurrent and Ca2+ concentration from the same cells. We show that for a limited number of measurements, for which the bleaching of the cone photopigment is too small to affect flash kinetics, the outer segment Ca2+ concentration closely follows the wave form of the flash response convolved with the dominant time constant for Ca2+ removal by Na+-Ca2+-K+ exchange. For a larger number of measurements, significant acceleration of the response kinetics by pigment bleaching inevitably occurs, but the Ca2+ concentration nevertheless rises and falls in approximate agreement with the flash wave form. During exposure to steady background light, the Ca2+ concentration falls in proportion to the steady-state current for dim backgrounds at all times and for bright backgrounds at steady state. At early times following the onset of bright backgrounds, however, the Ca2+ concentration is markedly higher than expected from the current of the cone. We show this to be the result of light-dependent Ca2+ release by bright background light, which can be abolished by pre-exposure of the cone to the membrane-permeant acetoxymethyl ester of the Ca2+ chelator BAPTA. Our results therefore demonstrate that the cone outer segment Ca2+ concentration is predominantly a function of the rate of influx and efflux of Ca2+ across the plasma membrane, but that a release of Ca2+ in bright light most probably from buffer sites within the cell can transiently elevate the Ca2+ concentration above the level expected from the open probability of the light-dependent channels. PMID- 17124272 TI - Kinetics of calcium spikes in rat cardiac myocytes. AB - The local calcium release flux signals (calcium spikes) evoked by membrane depolarization were recorded at high temporal resolution (2000 lines s(-1)) in isolated ventricular myocytes of male rats, using combination of scanning confocal microscopy and the patch-clamp technique. The kinetic properties of calcium spikes were investigated. The time course of calcium spike activation could be described reliably by a model with higher-order (n = 3) kinetics, but not by a first-order exponential process. A model of calcium spike with calcium release termination coupled to its activation was preferential to a model with the release termination independent of its activation. Three fluorescent calcium dyes (OG-5N, fluo-3, and fluo-4) were compared for calcium spike measurements. Experimental measurements as well as simulations showed that the occurrence and latency of calcium spikes could be measured faithfully with all indicators, while the kinetics of calcium spikes was reliably traced only with OG-5N. Calcium spikes evoked by a step depolarization from -50 to 0 mV commenced with a mean latency of 4.1 +/- 0.2 ms and peaked 6.7 +/- 0.2 ms later. Their full amplitudes were normally distributed. The activation time constant of calcium spikes was 3.1 +/- 0.1 ms, and the time constant of termination was 5.5 +/- 0.2 ms. A negative correlation was observed between the observed amplitude of calcium spikes and their time constant of activation, but there was no correlation between their observed amplitude and time constant of termination, in agreement with the concept of steep calcium-dependent activation and fateful inactivation of calcium release flux. PMID- 17124273 TI - High CO2 chemosensitivity versus wide sensing spectrum: a paradoxical problem and its solutions in cultured brainstem neurons. AB - CO2 central chemoreceptors play an important role in cardiorespiratory control. They are highly sensitive to P(CO2) in a broad range. These two sensing properties seem paradoxical as none of the known pH-sensing molecules can achieve both. Here we show that cultured neuronal networks are likely to solve the sensitivity versus spectrum problem with parallel and serial processes. Studies were performed on dissociated brainstem neurons cultured on microelectrode arrays. Recordings started after a 3 week initial period of culture. A group of neurons were dose-dependently stimulated by elevated CO2 with a linear response ranging from 20 to 70 Torr. The firing rate of some neurons increased by up to 30% in response to a 1 Torr P(CO2) change, indicating that cultured brainstem neuronal networks retain high CO2 sensitivity in a broad range. Inhibition of Kir channels selectively suppressed neuronal responses to hypocapnia and mild hypercapnia. Blockade of TASK channels affected neuronal response to more severe hypercapnia. These were consistent with the pKa values measured for these K+ channels in a heterologous expression system. The CO2 chemosensitivity was reduced but not eliminated by blockade of presynaptic input from serotonin, substance P or glutamate neurons, indicating that both pre and postsynaptic neurons contribute to the CO2 chemosensitivity. These results therefore strongly suggest that the physiological P(CO2) range appears to be covered by multiple sensing molecules, and that the high sensitivity may be achieved by cellular mechanisms via synaptic amplification in cultured brainstem neurons. PMID- 17124274 TI - Systemic, cellular and molecular analysis of chemoreflex-mediated sympathoexcitation by chronic intermittent hypoxia. AB - Patients with recurrent apnoeas exhibit autonomic abnormalities manifested as persistent increase in sympathetic nerve activity (SNA). Several studies suggest that chronic intermittent hypoxia (CIH) resulting from recurrent apnoeas is a major stimulus for evoking autonomic morbidity. Although it has been proposed that CIH, by way of activating the chemoreceptor reflex, leads to sympathetic excitation, the underlying mechanisms are incompletely understood. Studies on experimental models have provided new insights into the mechanisms associated with CIH-evoked sympathoexcitation. The purpose of this article is to highlight recent information on systemic, cellular and molecular analysis of the effects of CIH on chemoreceptor-mediated sympathoexcitation. Chronic intermittent hypoxia exerts two major effects on the chemoreceptor reflex: (a) augmentation of the carotid body and sympathetic effector responses to acute hypoxia; and (b) induction of long-lasting activation of both the sensor and the effector that persists several hours after termination of CIH. Available evidence indicates that CIH may facilitate processing of chemoreceptor afferent information at the central nervous system. Recent studies suggest that reactive oxygen species mediated signalling is a major cellular mechanism, and transcriptional activation by hypoxia-inducible factor-1 is one of the critical molecular mechanisms underlying chemoreceptor-mediated sympathoexcitation by CIH. PMID- 17124275 TI - Physiological basis for a causal relationship of obstructive sleep apnoea to hypertension. AB - Obstructive sleep apnoea (OSA) is causally related to systemic hypertension through sustained sympathoexcitation. The causes of this sympathoexcitation remain uncertain; however, substantial animal and human data suggest that cyclic intermittent hypoxia (CIH), as is experienced at night by patients with OSA, provides the causal link between upper airway obstruction during sleep and sympathetic activation during waking. Direct and indirect evidence indicates that CIH leads to sympathoexcitation by two mechanisms: (1) augmentation of peripheral chemoreflex sensitivity (hypoxic acclimatization); and (2) direct effects on sites of central sympathetic regulation, such as the subfornical organ and the paraventricular nucleus of the hypothalamus. Initial reports suggest that the molecular mechanisms influencing peripheral chemoreflex sensitivity and central sympathetic activity may be the same, involving such neuromodulators as angiotensin II, endothelin and nitric oxide. PMID- 17124276 TI - Intermittent hypoxia and vascular function: implications for obstructive sleep apnoea. AB - Obstructive sleep apnoea (OSA) has been implicated as a risk factor for the development of hypertension, stroke and myocardial infarction. The main cause of cardiovascular and cerebrovascular disease in OSA is thought to be exposure to intermittent hypoxia, which can lead to oxidative stress, inflammation, atherosclerosis, endothelial dysfunction and hypertension. These proposed mechanisms have been drawn from basic research in animal and human models of intermittent hypoxia in addition to clinical investigation of patients with OSA. This review outlines the association between OSA and vascular disease, describes basic mechanisms that may be responsible for this association and compares the results from studies of OSA subjects with those in experimental models of intermittent hypoxia. PMID- 17124277 TI - Looking beyond the obvious--a young woman with septic abortion and acute renal failure. PMID- 17124278 TI - Endophthalmitis, a rare metastatic bacterial complication of haemodialysis catheter-related sepsis. PMID- 17124279 TI - Stable T-cell reactivity after successful tapering of azathioprine in HLA identical living-related kidney transplant recipients despite minor histocompatibility antigen mismatches. AB - BACKGROUND: Human leukocyte antigen (HLA)-identical living-related (LR) kidney transplant recipients often receive the standard regimen of immunosuppression. We wondered whether these patients should be exposed to the side effects of these drugs any longer. Safe tapering of immunosuppression should not result in rejection and high donor-directed T-cell responses. In the present study, we investigated the effect of tapering azathioprine (AZA) on T-cell reactivity. METHODS: Fifteen HLA-identical LR kidney transplant recipients receiving a median of 150 mg/day AZA and 5-10 mg/day prednisone were tapered to a median of 50 mg/day AZA. Donor-, third-party and tetanus toxoid (TET)-reactivity were determined in interferon (IFN)-gamma and interleukin (IL)-13 Elispot assays, which reflect the T-helper (Th)1 and T-helper (Th)2 response. RESULTS: After the tapering of AZA, none of the patients developed acute rejection and the renal function remained stable, even at 1-year follow-up. The frequency of donor specific IFN-gamma and IL-13 producing cells (pc) was low. Tapering of AZA did not influence the frequency of both IFN-gamma and IL-13 pc. Also, the reactivity against third-party cells and TET remained unchanged. CONCLUSIONS: The AZA-dose can be safely reduced in recipients of an HLA-identical LR kidney transplant without affecting kidney function and without increasing T-cell responses directed against donor or other antigens. PMID- 17124280 TI - The periodontal status of pre-dialysis chronic kidney disease and maintenance dialysis patients. AB - BACKGROUND: Periodontitis contributes to generalized inflammation and development of systemic diseases, including atherosclerosis and cardiovascular disease. Its extent in maintenance haemodialysis (HD) patients is disputable and not known in continuous ambulatory peritoneal dialysis (CAPD) and pre-dialysis chronic kidney disease (CKD) patients. METHODS: One hundred and six patients (35 on HD, mean age, 56 years; 33 on CAPD, mean age 51 years; and 38 pre-dialysis CKD stage 2-5, mean age 51 years) from north-eastern Poland were enrolled. Dialysis subjects were recruited from a cohort of 141 HD and 61 CAPD patients. Two control groups comprised 26 generally healthy individuals with advanced periodontitis requiring specialized treatment, and 30 subjects from general population. Gingival index (GI), papillary bleeding index (PBI), plaque index (PI), loss of clinical attachment level (CAL) and community periodontal index of treatment needs (CPITN) were determined according to WHO recommendations. RESULTS: Average values of the indices in HD, CAPD, pre-dialysis CKD, advanced periodontitis and general population subjects were as follows: GI-1.37, 0.95, 1, 2 and 1; PBI-1.45, 0, 0, 2.20 and 1; PI-2.05, 1.59, 1, 2 and 1; and CAL loss-5.11, 3.47, 2.50, 4.68 and 1.40 mm, respectively. CPITN, analysed separately as community periodontal index and periodontal treatment needs, further indicated a high severity of periodontitis in all renal failure groups as compared with general population subjects. The disease was most advanced in maintenance HD patients-comparable to the full-symptomatic form of periodontitis; then it was successively diminished in CAPD and pre-dialysis CKD subjects. CONCLUSIONS: Periodontal disease is prevalent, severe and under recognized in renal failure patients. Prophylaxis and early dental treatment should be intensified in these subjects, and may be of interdisciplinary importance. PMID- 17124281 TI - Plasma exchange in the treatment of acute renal failure of myeloma. PMID- 17124282 TI - The case for ultraviolet light therapy in nephrogenic fibrosing dermopathy- report of two cases and review of the literature. PMID- 17124283 TI - Risedronate, an effective treatment for glucocorticoid-induced bone loss in CKD patients with or without concomitant active vitamin D (PRIUS-CKD). AB - BACKGROUND: Recent post hoc analysis proved the efficacy and tolerability of risedronate in osteoporotic patients with renal impairment, but the combination of active vitamin D in chronic kidney disease (CKD) patients taking glucocorticoids remains unknown. METHODS: We conducted a prospective study enrolling 114 CKD patients (creatinine clearance > or =30 ml/min/1.73 m(2)) receiving glucocorticoid therapy for > or =6 months. Eighty-eight subjects who had received active vitamin D (aVD) were randomly assigned to either a group treated with aVD only (group A), or to a group also receiving risedronate 2.5 mg/day (group B). The remaining patients (group C) received risedronate only. RESULTS: After 1 year 100 subjects were analysed. Risedronate was effective on the lumbar spine, but not on the femoral neck. The lumbar bone mineral density (BMD) significantly increased by 2.8 and 2.5% in groups B and C, respectively, but decreased by 1.0% in group A. Serum N-terminal telopeptides of type I collagen (S-NTX) and bone alkaline phosphatase (ALP) fell significantly in groups B and C at 3 and 6 months, respectively, while in group A S-NTX remained unchanged and bone ALP significantly increased. There was no significant difference between groups B and C regarding BMD and bone markers. The reduction rate of S-NTX (bone ALP) at 6 months predicted the increase in lumbar BMD at 1 year with a sensitivity of 73% (34%) and a specificity of 46.2% (100%). CONCLUSIONS: Risedronate is effective in increasing BMD with or without aVD in CKD patients receiving long-term glucocorticoid therapy. Bone markers are of some use in predicting the response to anti-resorptive therapy. PMID- 17124284 TI - Possible mechanisms explaining the tendency towards interstitial fibrosis in aristolochic acid-induced acute tubular necrosis. AB - BACKGROUND: We explored possible mechanisms responsible for the inability of plerosis and the tendency towards fibrosis in aristolochic acid-induced acute tubular necrosis (AA-ATN). METHODS: Renal biopsy tissues from eight AA-ATN cases were examined. Tubulointerstitial injury was semiquantitatively assessed. Immunohistochemical steptavidin-peroxide (SP) methods were used to determine the expressions of proliferating cell nuclear antigen (PCNA), epidermal growth factor (EGF), alpha-smooth muscle actin (alpha-SMA), transforming growth factor-beta(1) (TGF-beta(1)), connecting tissue growth factor (CTGF), fibronectin (FN), collagen III (Col-III), collagen IV (Col-IV), factor VIII-related antigen (VIII-Ag) and vascular endothelial growth factor (VEGF). Ultramicrostructure of endothelial cells and basement membrane of peritubular capillaries (PTC) and glomerular capillaries was detected by electron microscopy. These data were compared with that of 9 cases of antibiotic-induced ATN (a-ATN) and 10 cases of minor mesangioproliferative non-IgA glomerulonephritis, which served as a control group. RESULTS: In AA-ATN, almost no renal tubular epithelial cells (RTEC) were PCNA-positive (0.01 +/- 0.02%), and EGF expression was considerably decreased (9.55 +/- 7.22%). This was in contrast with the highly active tubular proliferation (PCNA-positive RTEC 47.25 +/- 19.33%, P < 0.05) and increased EGF expression in a-ATN (64.38 +/- 19.22%, P < 0.05). The expression of alpha-SMA in the tubulointerstitium, the number of interstitial TGF-beta(1)-positive cells and the CTGF-positive interstitial area were all increased in both a-ATN and AA-ATN, with no obvious differences between the two groups. With respect to extracellular matrix (ECM) deposition, FN, Col-III and Col-IV were detected only in the interstitium of AA-ATN. PTC lumina were decreased in size and misshapen in the AA ATN group. Also in AA-ATN, the luminal wall was partially disrupted, endothelial cells were swollen and vacuoles and granules were found in the cell plasma. Parts of the endothelial cells were detached from the tubular basement membrane. CONCLUSION: The strong ability for RTEC repair after acute injury was severely diminished in AA-ATN, and this effect may be partly due to reduced EGF expression. Anti-fibrosis mechanisms may also be impaired in AA-ATN, since both a ATN and AA-ATN had increased expression of TGF-beta(1) and CTGF, whereas only the latter group showed ECM deposition. Injury and loss of PTC occurred in AA-ATN, and this may contribute to tubulointerstitial damage, the inability of plerosis and the tendency towards fibrosis in this disease. PMID- 17124285 TI - Compensating occupationally related tenosynovitis and epicondylitis: a literature review. AB - OBJECTIVES: To assess occupational associations with tenosynovitis and epicondylitis, we conducted a systematic literature review. We focused particularly on evidence that might support compensation of these disorders 'on the balance of probabilities'. METHODS: We searched the MEDLINE and EMBASE electronic biomedical databases to 1 January 2005 using combinations of keyword and medical subject headings, and also the references cited in two state-of-the art reviews from the 1990s. Primary research reports were retrieved and checked for further relevant citations. From each paper, we abstracted a standardized set of information on study populations, exposure contrasts and estimates of effect. RESULTS: We found and summarized 18 papers. In the main, these based analysis on job titles rather than on directly assessed physical activities. Few occupations were studied more than once, however, and there was little consistent evidence of jobs or work activities that carried more than a doubling of risk for either disorder. CONCLUSION: Compensation of occupational illness can be problematic for disorders that are not specific to work and for which there are no distinctive clinical features in occupationally related cases. Attribution can, however, be made on the balance of probabilities if there is convincing evidence that risk is at least doubled in an occupational group. Our review highlights the relative lack of data to support such attribution for tenosynovitis and epicondylitis, and discusses the difficulty of compensating upper limb disorders. PMID- 17124286 TI - The sight of others' pain modulates motor processing in human cingulate cortex. AB - Neuroimaging evidence has shown that a network including cingulate cortex and bilateral insula responds to both felt and seen pain. Of these, dorsal anterior cingulate and midcingulate areas are involved in preparing context-appropriate motor responses to painful situations, but it is unclear whether the same holds for observed pain. Participants in this functional magnetic resonance imaging study viewed short animations depicting a noxious implement (e.g., a sharp knife) or an innocuous implement (e.g., a butter knife) striking a person's hand. Participants were required to execute or suppress button-press responses depending on whether the implements hit or missed the hand. The combination of the implement's noxiousness and whether it contacted the hand strongly affected reaction times, with the fastest responses to noxious-hit trials. Blood oxygen level-dependent signal changes mirrored this behavioral interaction with increased activation during noxious-hit trials only in midcingulate, dorsal anterior, and dorsal posterior cingulate regions. Crucially, the activation in these cingulate regions also depended on whether the subject made an overt motor response to the event, linking their role in pain observation to their role in motor processing. This study also suggests a functional topography in medial premotor regions implicated in "pain empathy," with adjacent activations relating to pain-selective and motor-selective components, and their interaction. PMID- 17124287 TI - Morphological, electrophysiological, and synaptic properties of corticocallosal pyramidal cells in the neonatal rat neocortex. AB - Neocortical pyramidal cells (PCs) project to various cortical and subcortical targets. In layer V, the population of thick tufted PCs (TTCs) projects to subcortical targets such as the tectum, brainstem, and spinal cord. Another population of layer V PCs projects via the corpus callosum to the contralateral neocortical hemisphere mediating information transfer between the hemispheres. This subpopulation (corticocallosally projecting cells [CCPs]) has been previously described in terms of their morphological properties, but less is known about their electrophysiological properties, and their synaptic connectivity is unknown. We studied the morphological, electrophysiological, and synaptic properties of CCPs by retrograde labeling with fluorescent microbeads in P13-P16 Wistar rats. CCPs were characterized by shorter, untufted apical dendrites, which reached only up to layers II/III, confirming previous reports. Synaptic connections between CCPs were different from those observed between TTCs, both in probability of occurrence and dynamic properties. We found that the CCP network is about 4 times less interconnected than the TTC network and the probability of release is 24% smaller, resulting in a more linear synaptic transmission. The study shows that layer V pyramidal neurons projecting to different targets form subnetworks with specialized connectivity profiles, in addition to the specialized morphological and electrophysiological intrinsic properties. PMID- 17124288 TI - Formation of a nematic fluid at high fields in Sr3Ru2O7. AB - In principle, a complex assembly of strongly interacting electrons can self organize into a wide variety of collective states, but relatively few such states have been identified in practice. We report that, in the close vicinity of a metamagnetic quantum critical point, high-purity strontium ruthenate Sr3Ru2O7 possesses a large magnetoresistive anisotropy, consistent with the existence of an electronic nematic fluid. We discuss a striking phenomenological similarity between our observations and those made in high-purity two-dimensional electron fluids in gallium arsenide devices. PMID- 17124289 TI - A sea-floor spreading event captured by seismometers. AB - Two-thirds of Earth's surface is formed at mid-ocean ridges, yet sea-floor spreading events are poorly understood because they occur far beneath the ocean surface. At 9 degrees 50'N on the East Pacific Rise, ocean-bottom seismometers recently recorded the microearthquake character of a mid-ocean ridge eruption, including precursory activity. A gradual ramp-up in activity rates since seismic monitoring began at this site in October 2003 suggests that eruptions may be forecast in the fast-spreading environment. The pattern culminates in an intense but brief (approximately 6-hour) inferred diking event on 22 January 2006, followed by rapid tapering to markedly decreased levels of seismicity. PMID- 17124290 TI - Structural basis for ribosome recruitment and manipulation by a viral IRES RNA. AB - Canonical cap-dependent translation initiation requires a large number of protein factors that act in a stepwise assembly process. In contrast, internal ribosomal entry sites (IRESs) are cis-acting RNAs that in some cases completely supplant these factors by recruiting and activating the ribosome using a single structured RNA. Here we present the crystal structures of the ribosome-binding domain from a Dicistroviridae intergenic region IRES at 3.1 angstrom resolution, providing a view of the prefolded architecture of an all-RNA translation initiation apparatus. Docking of the structure into cryo-electron microscopy reconstructions of an IRES-ribosome complex suggests a model for ribosome manipulation by a dynamic IRES RNA. PMID- 17124291 TI - Distinct populations of primary and secondary effectors during RNAi in C. elegans. AB - RNA interference (RNAi) is a phylogenetically widespread gene-silencing process triggered by double-stranded RNA. In plants and Caenorhabditis elegans, two distinct populations of small RNAs have been proposed to participate in RNAi: "Primary siRNAs" (derived from DICER nuclease-mediated cleavage of the original trigger) and "secondary siRNAs" [additional small RNAs whose synthesis requires an RNA-directed RNA polymerase (RdRP)]. Analyzing small RNAs associated with ongoing RNAi in C. elegans, we found that secondary siRNAs constitute the vast majority. The bulk of secondary siRNAs exhibited structure and sequence indicative of a biosynthetic mode whereby each molecule derives from an independent de novo initiation by RdRP. Analysis of endogenous small RNAs indicated that a fraction derive from a biosynthetic mechanism that is similar to that of secondary siRNAs formed during RNAi, suggesting that small antisense transcripts derived from cellular messenger RNAs by RdRP activity may have key roles in cellular regulation. PMID- 17124292 TI - Carbon trading. PMID- 17124293 TI - Global warming. U.N. conference puts spotlight on reducing impact of climate change. PMID- 17124294 TI - Developmental biology. Teams identify cardiac 'stem cell'. PMID- 17124295 TI - Fusion. Scientists reap ITER's first dividends. PMID- 17124296 TI - Sherwood Boehlert interview. Explaining science to power: make it simple, make it pay. Interview by Jeffrey Mervis. PMID- 17124297 TI - U.S. intellectual property. Patent experts hope high court will clarify what's obvious. PMID- 17124298 TI - U.S. intellectual property law. Government questions sequencing patent. PMID- 17124299 TI - U.S. science policy. Resignations rock Census Bureau. PMID- 17124300 TI - Chinese gene therapy. Splicing out the West? PMID- 17124301 TI - Chinese gene therapy. Gendicine's efficacy: hard to translate. PMID- 17124303 TI - Science funding. Italy's research crunch: election promises fade. PMID- 17124302 TI - Herve This profile. The joy of evidence-based cooking. PMID- 17124304 TI - Evolution. Two rapidly evolving genes spell trouble for hybrids. PMID- 17124305 TI - A debate over Iraqi death estimates. PMID- 17124306 TI - A nonprotein amino acid and neurodegeneration. PMID- 17124307 TI - Plants, RNAi, and the Nobel Prize. PMID- 17124308 TI - Comment on papers by Chong et al., Nishio et al., and Suri et al. on diabetes reversal in NOD mice. AB - Chong et al., Nishio et al., and Suri et al. (Reports, 24 March 2006, pp. 1774, 1775, and 1778) confirmed that treating nonobese diabetic (NOD) mice with an immune adjuvant and semisyngenic spleen cells can reverse the disease but found that spleen cells did not contribute to the observed recovery of pancreatic islets. We show that islet regeneration predominately originates from endogenous cells but that introduced spleen cells can also contribute to islet recovery. PMID- 17124309 TI - Mentoring. Volunteers bring passion to science outreach. PMID- 17124310 TI - Physics. Superconductivity with a twist. PMID- 17124311 TI - Neuroscience. The brain's dark energy. PMID- 17124312 TI - Plant science. Distributing nutrition. PMID- 17124313 TI - Atmosphere. Global change in the upper atmosphere. PMID- 17124314 TI - Paleoecology. Life's complexity cast in stone. PMID- 17124315 TI - What is natural? The need for a long-term perspective in biodiversity conservation. AB - Ecosystems change in response to factors such as climate variability, invasions, and wildfires. Most records used to assess such change are based on short-term ecological data or satellite imagery spanning only a few decades. In many instances it is impossible to disentangle natural variability from other, potentially significant trends in these records, partly because of their short time scale. We summarize recent studies that show how paleoecological records can be used to provide a longer temporal perspective to address specific conservation issues relating to biological invasions, wildfires, climate change, and determination of natural variability. The use of such records can reduce much of the uncertainty surrounding the question of what is "natural" and thereby start to provide important guidance for long-term management and conservation. PMID- 17124316 TI - Effective enforcement in a conservation area. AB - Wildlife within protected areas is under increasing threat from bushmeat and illegal trophy trades, and many argue that enforcement within protected areas is not sufficient to protect wildlife. We examined 50 years of records from Serengeti National Park in Tanzania and calculated the history of illegal harvest and enforcement by park authorities. We show that a precipitous decline in enforcement in 1977 resulted in a large increase in poaching and decline of many species. Conversely, expanded budgets and antipoaching patrols since the mid 1980s have greatly reduced poaching and allowed populations of buffalo, elephants, and rhinoceros to rebuild. PMID- 17124317 TI - A post-perovskite lens and D'' heat flux beneath the central Pacific. AB - Temperature gradients in a low-shear-velocity province in the lowermost mantle (D'' region) beneath the central Pacific Ocean were inferred from the observation of a rapid S-wave velocity increase overlying a rapid decrease. These paired seismic discontinuities are attributed to a phase change from perovskite to post perovskite and then back to perovskite as the temperature increases with depth. Iron enrichment could explain the occurrence of post-perovskite several hundred kilometers above the core-mantle boundary in this warm, chemically distinct province. The double phase-boundary crossing directly constrains the lowermost mantle temperature gradients. Assuming a standard but unconstrained choice of thermal conductivity, the regional core-mantle boundary heat flux (approximately 85 +/- 25 milliwatts per square meter), comparable to the average at Earth's surface, was estimated, along with a lower bound on global core-mantle boundary heat flow in the range of 13 +/- 4 terawatts. Mapped velocity-contrast variations indicate that the lens of post-perovskite minerals thins and vanishes over 1000 kilometers laterally toward the margin of the chemical distinct region as a result of a approximately 500-kelvin temperature increase. PMID- 17124318 TI - Ongoing buildup of refractory organic carbon in boreal soils during the Holocene. AB - Radiocarbon ages of vascular plant wax-derived n-alkanes preserved in well-dated Holocene sediments in an anoxic fjord (Saanich Inlet, Canada) were found to be not only substantially older than the depositional age but increasingly so during the Holocene. Assuming that n-alkanes serve as a proxy for recalcitrant terrigenous organic matter, this indicates that the accumulation of refractory organic carbon in soils that developed after the deglaciation of the American Pacific Northwest is ongoing and may still be far from equilibrium with mineralization and erosion rates. PMID- 17124319 TI - Abundance distributions imply elevated complexity of post-Paleozoic marine ecosystems. AB - Likelihood analyses of 1176 fossil assemblages of marine organisms from Phanerozoic (i.e., Cambrian to Recent) assemblages indicate a shift in typical relative-abundance distributions after the Paleozoic. Ecological theory associated with these abundance distributions implies that complex ecosystems are far more common among Meso-Cenozoic assemblages than among the Paleozoic assemblages that preceded them. This transition coincides not with any major change in the way fossils are preserved or collected but with a shift from communities dominated by sessile epifaunal suspension feeders to communities with elevated diversities of mobile and infaunal taxa. This suggests that the end Permian extinction permanently altered prevailing marine ecosystem structure and precipitated high levels of ecological complexity and alpha diversity in the Meso Cenozoic. PMID- 17124320 TI - Two Dobzhansky-Muller genes interact to cause hybrid lethality in Drosophila. AB - The Dobzhansky-Muller model proposes that hybrid incompatibilities are caused by the interaction between genes that have functionally diverged in the respective hybridizing species. Here, we show that Lethal hybrid rescue (Lhr) has functionally diverged in Drosophila simulans and interacts with Hybrid male rescue (Hmr), which has functionally diverged in D. melanogaster, to cause lethality in F1 hybrid males. LHR localizes to heterochromatic regions of the genome and has diverged extensively in sequence between these species in a manner consistent with positive selection. Rapidly evolving heterochromatic DNA sequences may be driving the evolution of this incompatibility gene. PMID- 17124321 TI - A NAC Gene regulating senescence improves grain protein, zinc, and iron content in wheat. AB - Enhancing the nutritional value of food crops is a means of improving human nutrition and health. We report here the positional cloning of Gpc-B1, a wheat quantitative trait locus associated with increased grain protein, zinc, and iron content. The ancestral wild wheat allele encodes a NAC transcription factor (NAM B1) that accelerates senescence and increases nutrient remobilization from leaves to developing grains, whereas modern wheat varieties carry a nonfunctional NAM-B1 allele. Reduction in RNA levels of the multiple NAM homologs by RNA interference delayed senescence by more than 3 weeks and reduced wheat grain protein, zinc, and iron content by more than 30%. PMID- 17124322 TI - Evolutionary history of Salmonella typhi. AB - For microbial pathogens, phylogeographic differentiation seems to be relatively common. However, the neutral population structure of Salmonella enterica serovar Typhi reflects the continued existence of ubiquitous haplotypes over millennia. In contrast, clinical use of fluoroquinolones has yielded at least 15 independent gyrA mutations within a decade and stimulated clonal expansion of haplotype H58 in Asia and Africa. Yet, antibiotic-sensitive strains and haplotypes other than H58 still persist despite selection for antibiotic resistance. Neutral evolution in Typhi appears to reflect the asymptomatic carrier state, and adaptive evolution depends on the rapid transmission of phenotypic changes through acute infections. PMID- 17124323 TI - Dissecting the functions of the mammalian clock protein BMAL1 by tissue-specific rescue in mice. AB - The basic helix-loop-helix (bHLH)-Per-Arnt-Sim (PAS) domain transcription factor BMAL1 is an essential component of the mammalian circadian pacemaker. Bmal1-/- mice lose circadian rhythmicity but also display tendon calcification and decreased activity, body weight, and longevity. To investigate whether these diverse functions of BMAL1 are tissue-specific, we produced transgenic mice that constitutively express Bmal1 in brain or muscle and examined the effects of rescued gene expression in Bmal1-/- mice. Circadian rhythms of wheel-running activity were restored in brain-rescued Bmal1-/- mice in a conditional manner; however, activity levels and body weight were lower than those of wild-type mice. In contrast, muscle-rescued Bmal1-/- mice exhibited normal activity levels and body weight yet remained behaviorally arrhythmic. Thus, Bmal1 has distinct tissue specific functions that regulate integrative physiology. PMID- 17124324 TI - A bacterial protein enhances the release and efficacy of liposomal cancer drugs. AB - Clostridium novyi-NT is an anaerobic bacterium that can infect hypoxic regions within experimental tumors. Because C. novyi-NT lyses red blood cells, we hypothesized that its membrane-disrupting properties could be exploited to enhance the release of liposome-encapsulated drugs within tumors. Here, we show that treatment of mice bearing large, established tumors with C. novyi-NT plus a single dose of liposomal doxorubicin often led to eradication of the tumors. The bacterial factor responsible for the enhanced drug release was identified as a previously unrecognized protein termed liposomase. This protein could potentially be incorporated into diverse experimental approaches for the specific delivery of chemotherapeutic agents to tumors. PMID- 17124325 TI - Predictive codes for forthcoming perception in the frontal cortex. AB - Incoming sensory information is often ambiguous, and the brain has to make decisions during perception. "Predictive coding" proposes that the brain resolves perceptual ambiguity by anticipating the forthcoming sensory environment, generating a template against which to match observed sensory evidence. We observed a neural representation of predicted perception in the medial frontal cortex, while human subjects decided whether visual objects were faces or not. Moreover, perceptual decisions about faces were associated with an increase in top-down connectivity from the frontal cortex to face-sensitive visual areas, consistent with the matching of predicted and observed evidence for the presence of faces. PMID- 17124326 TI - Electrophysiological response dynamics during focal cortical infarction. AB - While the intracellular processes of hypoxia-induced necrosis and the intercellular mechanisms of post-ischemic neurotoxicity associated with stroke are well documented, the dynamic electrophysiological (EP) response of neurons within the core or periinfarct zone remains unclear. The present study validates a method for continuous measurement of the local EP responses during focal cortical infarction induced via photothrombosis. Single microwire electrodes were acutely implanted into the primary auditory cortex of eight rats. Multi-unit neural activity, evoked via a continuous 2 Hz click stimulus, was recorded before, during and after infarction to assess neuronal function in response to local, permanent ischemia. During sham infarction, the average stimulus-evoked peak firing rate over 20 min remained stable at 495.5+/-14.5 spikes s-1, indicating temporal stability of neural function under normal conditions. Stimulus-evoked peak firing was reliably reduced to background levels (firing frequency in the absence of stimulus) following initiation of photothrombosis over a period of 439+/-92 s. The post-infarction firing patterns exhibited unique temporal degradation of the peak firing rate, suggesting a variable response to ischemic challenge. Despite the inherent complexity of cerebral ischemia secondary to microvascular occlusion, complete loss of EP function consistently occurred 300-600 s after photothrombosis. The results suggest that microwire recording during photothrombosis provides a simple and highly efficacious strategy for assessing the electrophysiological dynamics of cortical infarction. PMID- 17124327 TI - Toward a self-deploying shape memory polymer neuronal electrode. AB - The widespread application of neuronal probes for chronic recording of brain activity and functional stimulation has been slow to develop partially due to long-term biocompatibility problems with existing metallic and ceramic probes and the tissue damage caused during probe insertion. Stiff probes are easily inserted into soft brain tissue but cause astrocytic scars that become insulating sheaths between electrodes and neurons. In this communication, we explore the feasibility of a new approach to the composition and implantation of chronic electrode arrays. We demonstrate that softer polymer-based probes can be inserted into the olfactory bulb of a mouse and that slow insertion of the probes reduces astrocytic scarring. We further present the development of a micromachined shape memory polymer probe, which provides a vehicle to self-deploy an electrode at suitably slow rates and which can provide sufficient force to penetrate the brain. The deployment rate and composition of shape memory polymer probes can be tailored by polymer chemistry and actuator design. We conclude that it is feasible to fabricate shape memory polymer-based electrodes that would slowly self-implant compliant conductors into the brain, and both decrease initial trauma resulting from implantation and enhance long-term biocompatibility for long-term neuronal measurement and stimulation. PMID- 17124328 TI - Feature extraction for EEG-based brain-computer interfaces by wavelet packet best basis decomposition. AB - A method based on wavelet packet best basis decomposition (WPBBD) is investigated for the purpose of extracting features of electroencephalogram signals produced during motor imagery tasks in brain-computer interfaces. The method includes the following three steps. (1) Original signals are decomposed by wavelet packet transform (WPT) and a wavelet packet library can be formed. (2) The best basis for classification is selected from the library. (3) Subband energies included in the best basis are used as effective features. Three different motor imagery tasks are discriminated using the features. The WPBBD produces a 70.3% classification accuracy, which is 4.2% higher than that of the existing wavelet packet method. PMID- 17124329 TI - A silicon retina that reproduces signals in the optic nerve. AB - Prosthetic devices may someday be used to treat lesions of the central nervous system. Similar to neural circuits, these prosthetic devices should adapt their properties over time, independent of external control. Here we describe an artificial retina, constructed in silicon using single-transistor synaptic primitives, with two forms of locally controlled adaptation: luminance adaptation and contrast gain control. Both forms of adaptation rely on local modulation of synaptic strength, thus meeting the criteria of internal control. Our device is the first to reproduce the responses of the four major ganglion cell types that drive visual cortex, producing 3600 spiking outputs in total. We demonstrate how the responses of our device's ganglion cells compare to those measured from the mammalian retina. Replicating the retina's synaptic organization in our chip made it possible to perform these computations using a hundred times less energy than a microprocessor-and to match the mammalian retina in size and weight. With this level of efficiency and autonomy, it is now possible to develop fully implantable intraocular prostheses. PMID- 17124330 TI - An implantable neuroprosthesis for standing and walking in paraplegia: 5-year patient follow-up. AB - We present the results of a 5-year patient follow-up after implantation of an original neuroprosthesis. The system is able to stimulate both epimysial and neural electrodes in such a way that the complete flexor-extensor chain of the lower limb can be activated without using the withdrawal reflex. We demonstrate that standing and assisted walking are possible, and the results have remained stable for 5 years. Nevertheless, some problems were noted, particularly regarding the muscle response on the epimysial channels. Analysis of the electrical behaviour and thresholds indicated that the surgical phase is crucial because of the sensitivity of the functional responses to electrode placement. Neural stimulation proved to be more efficient and more stable over time. This mode requires less energy and provides more selective stimulation. This FES system can be improved to enable balanced standing and less fatiguing gait, but this will require feedback on event detection to trigger transitions between stimulation sequences, as well as feedback to the patient about the state of his lower limbs. PMID- 17124331 TI - Original electronic design to perform epimysial and neural stimulation in paraplegia. AB - This paper presents an original electronic architecture to manage epimysial and neural stimulation using the same implantable device. All the muscles needed to achieve lower limb movements such as standing and walking can thus be activated. Mainly for surgical reasons, some muscles need to be stimulated through different inputs: epimysium or motor nerve. We developed an electronic solution, including the design of an application-specific integrated circuit, to meet the requirements of both types of stimulation. Five years after the successful implantation of the system, we were able to evaluate the system's performance. The patient is still using the system at home and no failure occurred during this 5-year period. We conclude that the electronic design not only provides a unique investigative tool for research, but that it can also be used to restore the motor function of the lower limb. This technology has an advantage over external stimulation because the patient can safely use the system at home. However, improvements such as lower power consumption, and thus greater autonomy, are needed. We further conclude that the modelling of the electrical behaviour of the electrodes is reliable and the estimated parameter values are homogeneous and consistent for the same type of electrode. Thus, the three parameters of the first-order model can be identified from an acute animal experiment and provide a means to optimize the design of the output stage of implanted stimulators. PMID- 17124332 TI - Extraction of motor activity from the cervical spinal cord of behaving rats. AB - Injury at the cervical region of the spinal cord results in the loss of the skeletal muscle control from below the shoulders and hence causes quadriplegia. The brain-computer interface technique is one way of generating a substitute for the lost command signals in these severely paralyzed individuals using the neural signals from the brain. In this study, we are investigating the feasibility of an alternative method where the volitional signals are extracted from the cervical spinal cord above the point of injury. A microelectrode array assembly was implanted chronically at the C5-C6 level of the spinal cord in rats. Neural recordings were made during the face cleaning behavior with forelimbs as this task involves cyclic forelimb movements and does not require any training. The correlation between the volitional motor signals and the elbow movements was studied. Linear regression technique was used to reconstruct the arm movement from the rectified-integrated version of the principal neural components. The results of this study demonstrate the feasibility of extracting the motor signals from the cervical spinal cord and using them for reconstruction of the elbow movements. PMID- 17124333 TI - A new planar multielectrode array: recording from a rat auditory cortex. AB - In the study of the spatiotemporal properties of the cortex, a demand often arises for recording local field evoked potentials (LFEP) and neural spikes from a quantity of points at close range from each other. In such a situation a device composed of a lot of electrodes assembled in a single bunch would be suitable. Such circumstances gave us the impetus to create the device described in this paper, namely a new planar electrode array for in vivo multisite extracellular recording. The device is made of plastic and includes platinum electrodes 50 microm in diameter. The array of 64 incorporated microelectrodes is placed on the surface of the cortex of anesthetized rats. Recordings could be made through all electrodes for more than 1 h without damage to the cortex. The inter-polar distance is approximately 100 microm so that each individual electrode can record activity from a separate population of neurons near the cortical surface. The recording system described here is highly useful for visualizing spatiotemporal structure of the cortical activities and for imaging dynamic neuronal assemblies. PMID- 17124334 TI - A comparison of classification techniques for the P300 Speller. AB - This study assesses the relative performance characteristics of five established classification techniques on data collected using the P300 Speller paradigm, originally described by Farwell and Donchin (1988 Electroenceph. Clin. Neurophysiol. 70 510). Four linear methods: Pearson's correlation method (PCM), Fisher's linear discriminant (FLD), stepwise linear discriminant analysis (SWLDA) and a linear support vector machine (LSVM); and one nonlinear method: Gaussian kernel support vector machine (GSVM), are compared for classifying offline data from eight users. The relative performance of the classifiers is evaluated, along with the practical concerns regarding the implementation of the respective methods. The results indicate that while all methods attained acceptable performance levels, SWLDA and FLD provide the best overall performance and implementation characteristics for practical classification of P300 Speller data. PMID- 17124335 TI - Bio-heat transfer model of deep brain stimulation-induced temperature changes. AB - There is a growing interest in the use of chronic deep brain stimulation (DBS) for the treatment of medically refractory movement disorders and other neurological and psychiatric conditions. Fundamental questions remain about the physiologic effects of DBS. Previous basic research studies have focused on the direct polarization of neuronal membranes by electrical stimulation. The goal of this paper is to provide information on the thermal effects of DBS using finite element models to investigate the magnitude and spatial distribution of DBS induced temperature changes. The parameters investigated include stimulation waveform, lead selection, brain tissue electrical and thermal conductivities, blood perfusion, metabolic heat generation during the stimulation and lead thermal conductivity/heat dissipation through the electrode. Our results show that clinical DBS protocols will increase the temperature of surrounding tissue by up to 0.8 degrees C depending on stimulation/tissue parameters. PMID- 17124336 TI - Nanoscale laminin coating modulates cortical scarring response around implanted silicon microelectrode arrays. AB - Neural electrodes could significantly enhance the quality of life for patients with sensory and/or motor deficits as well as improve our understanding of brain functions. However, long-term electrical connectivity between neural tissue and recording sites is compromised by the development of astroglial scar around the recording probes. In this study we investigate the effect of a nanoscale laminin (LN) coating on Si-based neural probes on chronic cortical tissue reaction in a rat model. Tissue reaction was evaluated after 1 day, 1 week, and 4 weeks post implant for coated and uncoated probes using immunohistochemical techniques to evaluate activated microglia/macrophages (ED-1), astrocytes (GFAP) and neurons (NeuN). The coating did not have an observable effect on neuronal density or proximity to the electrode surface. However, the response of microglia/macrophages and astrocytes was altered by the coating. One day post implant, we observed an approximately 60% increase in ED-1 expression near LN coated probe sites compared with control uncoated probe sites. Four weeks post implant, we observed an approximately 20% reduction in ED-1 expression along with an approximately 50% reduction in GFAP expression at coated relative to uncoated probe sites. These results suggest that LN has a stimulatory effect on early microglia activation, accelerating the phagocytic function of these cells. This hypothesis is further supported by the increased mRNA expression of several pro inflammatory cytokines (TNF-alpha, IL-1 and IL-6) in cultured microglia on LN bound Si substrates. LN immunostaining of coated probes immediately after insertion and retrieval demonstrates that the coating integrity is not compromised by the shear force during insertion. We speculate, based on these encouraging results, that LN coating of Si neural probes could potentially improve chronic neural recordings through dispersion of the astroglial scar. PMID- 17124337 TI - Computational model of a primate arm: from hand position to joint angles, joint torques and muscle forces. AB - Three-dimensional reaching by non-human primates is an important behavioral paradigm for investigating representations existing in motor control areas of the brain. Most studies to date have correlated neural activity to a few of the many arm motion parameters including: global hand position or velocity, joint angles, joint angular velocities, joint torques or muscle activations. So far, no single study has been able to incorporate all these parameters in a meaningful way that would allow separation of these often highly correlated variables. This paper introduces a three-dimensional, seven degree-of-freedom computational musculoskeletal model of the macaque arm that translates the coordinates of eight tracking markers placed on the arm into joint angles, joint torques, musculotendon lengths and finally into an optimized prediction of muscle forces. This paper uses this model to illustrate how the classic center-out reaching task used by many researchers over the last 20 years is not optimal in separating out intrinsic, extrinsic, kinematic and kinetic variables. However, by using the musculoskeletal model to design and test novel behavioral movement tasks, a priori, it is possible to disassociate the myriad of movement parameters in motor neurophysiological reaching studies. PMID- 17124338 TI - Removal of ocular artifacts from EEG using adaptive thresholding of wavelet coefficients. AB - Electroencephalogram (EEG) gives researchers a non-invasive way to record cerebral activity. It is a valuable tool that helps clinicians to diagnose various neurological disorders and brain diseases. Blinking or moving the eyes produces large electrical potential around the eyes known as electrooculogram. It is a non-cortical activity which spreads across the scalp and contaminates the EEG recordings. These contaminating potentials are called ocular artifacts (OAs). Rejecting contaminated trials causes substantial data loss, and restricting eye movements/blinks limits the possible experimental designs and may affect the cognitive processes under investigation. In this paper, a nonlinear time-scale adaptive denoising system based on a wavelet shrinkage scheme has been used for removing OAs from EEG. The time-scale adaptive algorithm is based on Stein's unbiased risk estimate (SURE) and a soft-like thresholding function which searches for optimal thresholds using a gradient based adaptive algorithm is used. Denoising EEG with the proposed algorithm yields better results in terms of ocular artifact reduction and retention of background EEG activity compared to non-adaptive thresholding methods and the JADE algorithm. PMID- 17124339 TI - Biomarkers in multiple sclerosis. PMID- 17124340 TI - Multiple sclerosis cerebrospinal fluid biomarkers. AB - Cerebrospinal fluid (CSF) is the body fluid closest to the pathology of multiple sclerosis (MS). For many candidate biomarkers CSF is the only fluid that can be investigated. Several factors need to be standardized when sampling CSF for biomarker research: time/volume of CSF collection, sample processing/storage, and the temporal relationship of sampling to clinical or MRI markers of disease activity. Assays used for biomarker detection must be validated so as to optimize the power of the studies. A formal method for establishing whether or not a particular biomarker can be used as a surrogate end-point needs to be adopted. This process is similar to that used in clinical trials, where the reporting of studies has to be done in a standardized way with sufficient detail to permit a critical review of the study and to enable others to reproduce the study design. A commitment must be made to report negative studies so as to prevent publication bias. Pre-defined consensus criteria need to be developed for MS-related prognostic biomarkers. Currently no candidate biomarker is suitable as a surrogate end-point. Bulk biomarkers of the neurodegenerative process such as glial fibrillary acidic protein (GFAP) and neurofilaments (NF) have advantages over intermittent inflammatory markers. PMID- 17124341 TI - NMO-IgG: a specific biomarker for neuromyelitis optica. AB - Neuromyelitis optica (NMO) is an inflammatory demyelinating disease that principally targets the optic nerves and spinal cord and often leads to severe disability and occasionally life threatening respiratory failure. Although its clinical manifestations overlap with those of multiple sclerosis (MS), in established cases these two conditions can be distinguished on the basis of clinical, radiological, and routine spinal fluid studies. The diagnosis in early cases or limited forms of NMO is difficult. We recently discovered a unique IgG autoantibody (NMO-IgG) that is highly specific to patients with NMO and thus a valuable diagnostic aid. Its antigen, aquaporin-4 (AQP4), is the central nervous system's predominant water channel protein. This antibody has not yet been proven to be pathogenic, but several facts suggest that it might be, including the similarity of the immunohistochemical pattern of NMO-(AQP4) IgG binding to mouse CNS tissues to the pattern of immune complex deposition in autopsied patients' spinal cord tissue. The spectrum of diseases identified by NMO-IgG is broader than has previously been recognized clinically and includes incomplete forms of NMO, such as recurrent transverse myelitis without optic neuritis and recurrent optic neuritis without myelitis. PMID- 17124342 TI - Biomarkers in multiple sclerosis: role of antibodies. AB - The first international workshop on "Biomarkers in Multiple Sclerosis" was organized by B. Bielekova, R. Hohlfeld, R. Martin and U. Utz from April 14-16, 2004, in Washington, DC. The workshop intended to discuss the current status and potential applicability of biological markers for the understanding of the pathogenesis, diagnosis, and therapy of multiple sclerosis. The present review summarizes the presentation on the potential role of antibodies as biomarkers for diagnosis, disease activity, classification and prediction of clinical courses in multiple sclerosis. PMID- 17124343 TI - Comprehensive phenotyping in multiple sclerosis: discovery based proteomics and the current understanding of putative biomarkers. AB - Currently, there is no single test for multiple sclerosis (MS). Diagnosis is confirmed through clinical evaluation, abnormalities revealed by magnetic resonance imaging (MRI), and analysis of cerebrospinal fluid (CSF) chemistry. The early and accurate diagnosis of the disease, monitoring of progression, and gauging of therapeutic intervention are important but elusive elements of patient care. Moreover, a deeper understanding of the disease pathology is needed, including discovery of accurate biomarkers for MS. Herein we review putative biomarkers of MS relating to neurodegeneration and contributions to neuropathology, with particular focus on autoimmunity. In addition, novel assessments of biomarkers not driven by hypotheses are discussed, featuring our application of advanced proteomics and metabolomics for comprehensive phenotyping of CSF and blood. This strategy allows comparison of component expression levels in CSF and serum between MS and control groups. Examination of these preliminary data suggests that several CSF proteins in MS are differentially expressed, and thus, represent putative biomarkers deserving of further evaluation. PMID- 17124344 TI - Chemokine receptors as biomarkers in multiple sclerosis. AB - Leukocyte infiltrates characterize tissue inflammation and are thought to be integral in the pathogenesis of multiple sclerosis (MS). This attribute underlines the importance of understanding mechanisms of leukocyte migration. Chemokines are secreted proteins which govern leukocyte trafficking into targeted organs. Chemokine receptors (CKR) are differentially expressed on leukocytes and their modulation is a potential target for MS disease modifying therapies. Chemokines and their receptors are also potential biomarkers of both disease activity and response to treatment. We describe the fluctuations in CKR expression on peripheral leukocytes in a group of MS patients followed longitudinally for up to 36 months. We observed little fluctuation in CKR expression within each patient over time, despite considerable variability in CKR expression between patients. These observations suggest that individual patients have a CKR set point, and this set point varies from one patient to another. Evaluation of chemokines or chemokine receptors as biomarkers in MS will need to account for this individual variability in CKR expression. PMID- 17124345 TI - Biomarkers indicative of blood-brain barrier disruption in multiple sclerosis. AB - Blood-brain barrier (BBB) disruption is one of the hallmarks of multiple sclerosis (MS). It is incompletely understood whether BBB disruption is the initial MS event leading to MS lesion formation or whether it is merely a consequence of cellular infiltration in the central nervous system (CNS). The presence of gadolinium enhancing (Gd+) lesions on serial brain MRI scans is frequently used to evaluate BBB disruption. The presence of Gd enhancement has therefore been used as a reference for most works evaluating promising biomarkers of BBB disruption that are reviewed here. These promising biomarkers include cytokines and chemokines, and their receptors, cell surface markers, and matrix metalloproteinases and their natural inhibitors. At this time, none of these markers have been shown as sensitive as the presence of Gd enhancement to reflect BBB disruption. However, MRI scanning is not only unpractical and expensive; it may also under represent the overall extent of BBB disruption. Developing new MS biomarkers that are sensitive and specific for BBB disruption could 1) improve the monitoring of disease activity; 2) improve the monitoring of response to MS therapies which target BBB disruption; and 3) advance our understanding of dynamic MS processes participating in BBB disruption. PMID- 17124346 TI - Biomarkers that discriminate multiple myeloma patients with or without skeletal involvement detected using SELDI-TOF mass spectrometry and statistical and machine learning tools. AB - Multiple Myeloma (MM) is a severely debilitating neoplastic disease of B cell origin, with the primary source of morbidity and mortality associated with unrestrained bone destruction. Surface enhanced laser desorption/ionization time of-flight mass spectrometry (SELDI-TOF MS) was used to screen for potential biomarkers indicative of skeletal involvement in patients with MM. Serum samples from 48 MM patients, 24 with more than three bone lesions and 24 with no evidence of bone lesions were fractionated and analyzed in duplicate using copper ion loaded immobilized metal affinity SELDI chip arrays. The spectra obtained were compiled, normalized, and mass peaks with mass-to-charge ratios (m/z) between 2000 and 20,000 Da identified. Peak information from all fractions was combined together and analyzed using univariate statistics, as well as a linear, partial least squares discriminant analysis (PLS-DA), and a non-linear, random forest (RF), classification algorithm. The PLS-DA model resulted in prediction accuracy between 96-100%, while the RF model was able to achieve a specificity and sensitivity of 87.5% each. Both models as well as multiple comparison adjusted univariate analysis identified a set of four peaks that were the most discriminating between the two groups of patients and hold promise as potential biomarkers for future diagnostic and/or therapeutic purposes. PMID- 17124347 TI - DHPLC screening of ATM gene in Italian patients affected by ataxia telangiectasia: fourteen novel ATM mutations. AB - The gene for ataxia-telangiectasia (A-T:MIM: #208900), ATM, spans about 150 kb of genomic DNA and is composed of 62 coding exons. ATM mutations are found along the entire coding sequence of the gene, without evidence of mutational hot spots. Using DNA as the starting material, we used denaturing high performance liquid chromatography (DHPLC) technique to search for ATM gene mutations. Initially, DHPLC was validated in a retrospective study of 16 positive control samples that included 19 known mutations; 100% of mutations were detected. Subsequently, DHPLC was used to screen for mutations a cohort of 22 patients with the classical form of A-T. A total of 27 different mutations were identified on 38 of the 44 alleles, corresponding to a 86% detection rate. Fourteen of the mutations were novel. In addition, 15 different variants and polymorphisms of unknown functional significance were found. The high incidence of new and individual A-T mutations in our cohort of patients demonstrates marked mutational heterogeneity of A-T in Italy and corroborate the efficiency of DHPLC as a method for the mutation screening of A-T patients. PMID- 17124348 TI - Expression of telomerase reverse transcriptase in psoriatic lesional skin. AB - BACKGROUND: Little information is known about telomerase expression in the chronic benign hyperproliferative skin disease, psoriasis. Further studies are still required to investigate its usefulness as a biomarker of this skin disorder. AIM: To investigate the expression of human telomerase reverse transcriptase (hTERT) in psoriatic lesional skin and its relation to disease severity. METHODS: The levels of hTERT-mRNA were quantified using real time RT PCR in lesional versus nonlesional skin specimens from 24 psoriatic patients. RESULTS: The expression of hTERT was detected in 16 psoriatic lesional specimens (66.7%), but in none of the normal skin. There was no relation between hTERT expression level and age of the patient or the duration of the disease. Among hTERT-positive patients, a significant positive correlation was observed between hTERT-mRNA levels and both the Psoriasis Area-and-Severity Index (PASI) and scaling scores (p = 0.012 & p = 0.006, respectively). CONCLUSION: Telomerase mRNA is detectable in lesional skin of most psoriatic patients and correlates with the severity of the disease and the rate of epidermal proliferation. PMID- 17124349 TI - [Management and development of the dangerous preparation archive]. AB - In the year 2000 an archive of dangerous preparations was created at the National Health Institute (Istituto Superiore di Sanita), following a principle included in the Directive 88/379/EEC on dangerous preparations, subsequently modified by the Directive 1999/45/EC, concerning the creation of a data bank on dangerous preparations in each European country. The information stored in the archive is useful for purposes of health consumer's and workers protection and prevention, and particularly in case of acute poisonings. The archive is fully informatised, therefore the companies can send the information using the web and the authorized Poison Centres can find the information on the archive using the web. In each Member State different procedures are in place to comply with the 1999/45/EC Directive; therefore an international coordination could be useful in order to create an European network of national data-banks on dangerous preparations. PMID- 17124350 TI - [Profile of acute poisoning in Italy. Analysis of the data reported by Poison Centres]. AB - Information relative to about 400 000 cases of human intoxications, registered by nine Italian Poison Centres between 1991 and 1998 is presented. Data have been collected and elaborated in the framework of an European project on improving the prevention and treatment of acute human poisoning (90/C 329/EEC Resolution). Sex, age group, etiological agent, place and circumstances of poisoning and risk estimation are the parameters analyzed for the characterization of this phenomenon. The following conclusions can be summarized from the overall picture. There is a slight prevalence of males over females (50.0% against 45.7%); 1-4 year age group presents the highest risk (37.0%), followed by 20-49 group (25.8%); drugs and household products are the prevalent causes of intoxications (37.4% and 26.0%, respectively); home is the place where 84.9% of accidents occur. Poisoning is accidental in 73.5% of the cases whilst suicides represent 18.7%. However, the outcome is positive in almost all cases and fatal accidents are not reported in the present casuistry. PMID- 17124351 TI - [Accreditation criteria and quality standards for Poisons centres: development of a quality management system within the Milan Poisons centre]. AB - Poisons centres throughout Italy and Europe vary considerably in terms of their institutions and organisation. The European Association of Poisons Centres and Clinical Toxicologists (EAPCCT) has laid down the activities that a poisons centre must carry out, specifying minimum and maximum standards required. These directions allow an evaluation of the service provided. In 2002 Milan Poisons Centre began a project aiming to introduce concepts and methodology proper of the quality systems within poisons centres' institutional activity. Concluded, the project resulted in the centre's certification and the documentation of its procedures: this may now contribute to help define the status and activity of poisons centres in Italy. PMID- 17124352 TI - [Development of an informatic model for monitoring and clinical and epidemiological evaluation of acute intoxication in Emergency Departments in Italy]. AB - Data collection at Emergency Departments (ED), especially with regard to toxicological aspects, is usually incomplete; they further deteriorate following the recent diffusion of data processing and retrieval systems, possibly different from region to region. The present paper deals with the setting of a computer program, specific for toxicological patients, easy to access from the general data collection system. PMID- 17124353 TI - [Acute intoxications and poisonings in Italian Emergency Rooms]. AB - The analysis of 2402 prospective records of subjects consecutively attending eleven Emergency Rooms located in Northern, Central and Southern Italy, during the November 2002-May 2003 period, because of acute intoxication and poisoning (AI), shows that 10 admissions per 1000 are due to AI, accounting at national level for over 240,000 yearly admissions. Six AI of ten are caused by alcohol, alone or in combination with drugs and other poisonings, 2 by drugs and 2 by other types of AI. AI are more frequent among males (65%) and 20-40 ages. AI due to alcohol and drugs are more typical of young males, living in Central and Northern Italy, while those due to medicines are proportionally more represented among females of Southern Italy. PMID- 17124354 TI - [Antidotes availability in Emergency Departments of the Italian National Health System and development of a national data-bank on antidotes]. AB - The availability of antidotes in Italian hospitals has been evaluated through the answers to a specific questionnaire sent to all Italian Emergency Departments, Intensive Care Units, 118 emergency response system, and Poison Centres. Five Poison Centres and, approximately, the 30% of the Emergency Departments and Intensive Care Units of all Italian emergency hospitals answered to the questionnaire. The results point out an insufficient availability of antidotes in the Italian emergency hospitals, with an almost total absence of those necessary for the treatment of less frequent and less known poisonings (e.g. digoxin, industrial agents), also when the antidote is a lifesaving drug. To improve the antidotes availability for the toxicological emergencies and to facilitate its supplying, a "national antidotes data-base" (BaNdA) has been realized, freely available to the hospital services which register themselves and make their antidotes stockpile available. PMID- 17124355 TI - [A relational database to store Poison Centers calls]. AB - Italian Poison Centers answer to approximately 100,000 calls per year. Potentially, this activity is a huge source of data for toxicovigilance and for syndromic surveillance. During the last decade, surveillance systems for early detection of outbreaks have drawn the attention of public health institutions due to the threat of terrorism and high-profile disease outbreaks. Poisoning surveillance needs the ongoing, systematic collection, analysis, interpretation, and dissemination of harmonised data about poisonings from all Poison Centers for use in public health action to reduce morbidity and mortality and to improve health. The entity-relationship model for a Poison Center relational database is extremely complex and not studied in detail. For this reason, not harmonised data collection happens among Italian Poison Centers. Entities are recognizable concepts, either concrete or abstract, such as patients and poisons, or events which have relevance to the database, such as calls. Connectivity and cardinality of relationships are complex as well. A one-to-many relationship exist between calls and patients: for one instance of entity calls, there are zero, one, or many instances of entity patients. At the same time, a one-to-many relationship exist between patients and poisons: for one instance of entity patients, there are zero, one, or many instances of entity poisons. This paper shows a relational model for a poison center database which allows the harmonised data collection of poison centers calls. PMID- 17124356 TI - [Peculiar features of childhood poisoning and in the maternal-fetal period]. AB - Acute toxic exposures in childhood are quite a frequent event in Paediatric Emergency Medicine. Despite that, there are few papers about clinical toxicology in children, at least in Italy. This paper is the first that takes into account both epidemiological aspects and clinical features of acute poisoning in children on a national basis. Collected data show the increased use of activated charcoal as a treatment and the use of the short stay observation unit as an appropriate answer to the real clinical situation, although some aspects are still controversial. It will be necessary to reconsider the whole matter widening the number of participating centres, so that the most controversial aspects may be clarified. PMID- 17124357 TI - [Development and first implementation of an instrument to measure the social relations network in a town]. AB - A programme of personal relationships improvement is under way in the town of Trento in Northern Italy. It was decided to carry out a survey to describe the frequency and quality of interpersonal links as a baseline. A questionnaire to assess frequency and quality of interpersonal relationships has been validated (QRI) and a first description of its distribution in a representative sample of Trento population has been achieved. The questionnaire has some interesting original features, in particular it investigates the relationships with relatives in the same ways as with others, gives special attention to neighbours and asks separate judgements about the quality of relationships with different kind of people. The questionnaire gives important information which are not easy to find. In Trento interpersonal relationships seem good, especially for the possibility to receive practical help. Neighbourhood networks can certainly be improved. Only comparisons with other settings and in time can allow more solid conclusions. The English version of the questionnaire is available through the first author. PMID- 17124358 TI - [Polyphenols and endogenous antioxidant defences: effects on glutathione and glutathione related enzymes]. AB - Among diet antioxidants, polyphenols, naturally occurring in vegetables, fruits and plant-derived beverages such as tea, red wine, and extra virgin olive oil, are the most abundant ones. In vitro cell culture experiments have shown that polyphenols possess antioxidant properties, and it is thought that these activities can contribute to the prevention of several oxidative stress associated diseases. It has however become clear that the mechanisms of action of polyphenols go beyond their intrinsic reducing capabilities, being able to exert other additional effects that are as yet poorly understood. This article gives an overview of the most recent data on the subject and describe the additional functions that polyphenols can have in biological systems, focusing on their effects on glutathione and its related enzymes. Evidence is provided of a tight connection between exogenous and endogenous antioxidants that appear to act in a coordinated fashion. Experimental data indicate that polyphenols may offer an indirect protection by activating endogenous defense systems. It is reasonable to hypothesize that this is achieved, at least in part, through antioxidant responsive elements (ARE) present in the promoter regions of many of the genes inducible by oxidative and chemical stress. The latest studies strongly suggest that dietary polyphenols can stimulate antioxidant enzyme transcription through ARE. PMID- 17124359 TI - [A survey on some Italian doctors' opinions about errors in clinical medicine]. AB - The paper discusses opinions on medical errors from the scientific literature and from a survey on 173 medical doctors working in a large hospital (> 500 beds) in Rome (Italy). The study is meant to explore doctors' opinions on professional and/or system responsibility in front of errors. In our hypothesis doctors are more favourable to attribute responsibility to individual carers rather than to institutions, and they are interested in solutions involving relational and communicational enhancements more than in technological or systemic innovations for risk management. The focus of the questionnaire is on errors' frequency, their principal causes and possible remedies, and their emotional impact. The main findings of the survey are presented. They confirm the idea of a medical culture essentially focused on individual responsibility rather than on system's responsibility. According to doctors the major causes of errors result from work overload and from a lack of communication inside medical teams and among different medical specialties. System errors, as well as technological solutions, are more quoted by doctors working in laboratories. Psychological consequences of errors vary by doctors' sex, length of exercise, and place of activity. PMID- 17124360 TI - [Formaldehyde in air of indoor and outdoor environments of urban area, relationships man's exposure]. AB - This study presents the results of man's exposure to formaldehyde, carried out in indoor and outdoor environments in the city of Ferrara. The processes were obtained on average concentration measurements carried out for one month by indoor, outdoor and personal samplings. Concentration values obtained by personal samplings (PE) were compared to both outdoor and indoor values, the latter including domestic and working environments. The results showed that there was a significant difference between formaldeyde concentrations outdoor and PE (p = 0,03 < alpha = 0,05), while a significant difference was not found between indoor and PE. Indoor values (values as average of 38 samples) were 19,5 microg/m3, outdoor ones were 6,9 microg/m3 and PE 19,9 microg/m3. The average indoor value in domestic environments (20,7 microg/m3) was higher than the value found in working environments (17,9 microg/m3). Binary comparisons showed that indoor and outdoor, together with indoor and personal samplings, display the same variability of formaldehyde concentrations. PMID- 17124361 TI - Mercury emission from crematoria. AB - The purpose of this study, undertaken at a cremator representing an example of current equipment and cremation practices in use in Italy, is to assess the possible mercury emitted during cremation and substantiate the current data available. This paper reports some preliminary results concerning mercury and total particulate matter emissions during three cremation processes. The obtained results gave a mercury concentration ranging from 0.005 to 0.300 mg/m3 and a mercury emission factor ranging from 0.036 to 2.140 g/corpse cremated. The total particulate matter concentration range was 1.0 to 2.4 mg/m3. PMID- 17124362 TI - Medical treatment of benign insulinoma using octreotide LAR: a case report. AB - In some patients with insulinoma, surgery is not possible due to either difficulties in detecting the tumor or advanced age. These patients need medical treatment for hypoglycemia. We report a case of benign insulinoma using the long acting octreotide formulation, octreotide long-acting repeatable (octreotide LAR), as a medical therapy. A 67-year-old woman was referred to our hospital for examinations of hypoglycemia. A blood sample taken during a hypoglycemic episode revealed low plasma glucose concentration, hyperinsulinemia and a high C-peptide level. An abdominal CT scan demonstrated a hypervascular tumor in the body of pancreas. She was diagnosed with insulinoma. As the patient refused surgical resection of the pancreas tumor, we started to use the somatostatin analogue, octreotide, for treatment of hypoglycemia. After the treatment her plasma glucose levels were elevated and serum immunoreactive insulin (IRI) levels were decreased. For long-term treatment, we changed the treatment from daily subcutaneous injection of octreotide to monthly intramuscular administration of octreotide LAR. This treatment was also effective and hypoglycemic attacks disappeared. Both plasma glucose levels and serum IRI levels were improved. Our case demonstrated that octreotide LAR was useful for long-term medical treatment of insulinoma. PMID- 17124363 TI - LEPR, ADBR3, IRS-1 and 5-HTT genes polymorphisms do not associate with obesity. AB - Obesity is a growing problem and is associated with numerous medical conditions. In several genes coding for molecules involved in the regulation of body weight (fat mass) and thermogenesis, polymorphisms have been reported which possibly modify human obesity risk. The aim of this study was to determine the incidence of the following polymorphisms in the following genes in 262 obese (BMI > or = 30) and 138 control (BMI < or = 25) subjects: leptin receptor (LEPR)-Gln223Arg, B3-adrenergic receptor (B3-AR)-Trp64Arg, serotonin transporter (5-HTT)--a 44-base pair insertion/deletion functional polymorphism in the 5-HTTLPR and insulin receptor substrate-1 (IRS-1)-Gly972Arg. Our hypothesis was that these polymorphisms would occur more frequently in the obese population. The polymorphisms were determined by polymerase chain reaction (PCR) and restriction genotyping in study population. In our results, no strong associations were observed between BMI status and these polymorphisms. Weak, though significant, association coefficients obtained with HTT and LEPR loci indicate that the genotype numbers at these loci may depend on BMI status to some extent. PMID- 17124364 TI - Prevalence of primary aldosteronism: should we screen for primary aldosteronism before treating hypertensive patients with medication? AB - The present review examines various reports describing prevalence of primary aldosteronism (PA) among hypertensives and the screening method of PA to better understand the current concepts used for diagnosing and managing PA among clinicians as well as specialists. Here, we describe and compare the prevalence of PA in Japan, which is well known to induce various vascular complications due to hyperaldosteronemia, resulting in cerebral infarction, myocardial infarction and renal failure, with that in another Asian area, US, European countries, Australia and Africa. The incidence rates for PA among hypertensives were recently reported to be widely raged between 3.2% and 20%. Those discrepancies are due in part to the completely different characteristics of the starting subjects used for studying the prevalence. Moreover, the criteria for screening PA among hypertensives, including aldosterone-renin ratio (ARR), and confirmatory tests for definitely diagnosing PA, such as saline infusion test are varied. We had already reported that a diagnosis of PA was made in 61 (6%) of the 1,020 hypertensive patients during the past five years, from 1995 until 1999. In our study, only 18% of the patients showed a serum K level of 3.3 mEq/l or less. Thus, many clinicians seem to misdiagnose PA as essential hypertension, because of absence of hypokalemia. Finally, we describe highlight key evidence for optimal methods for screening and definitely diagnosing PA among hypertensive patients in order to avoid misjudgment before or after treating hypertensive patients. PMID- 17124365 TI - Activation of c-Jun NH2-terminal kinase during islet isolation. AB - Pancreatic islet transplantation has been remarkably improved by the Edmonton protocol; however, it is not easy to achieve insulin independence after islet transplantation from one donor pancreas. The islet isolation procedure itself destroys cellular and noncellular components of the pancreas that probably play a role in supporting islet survival. Further islet transplantation exposes cells to a variety of stressful stimuli such as proinflammatory cytokines. The reduction in islet mass immediately after isolation and transplantation implicates beta cell death by apoptosis and the prerecruitment of intracellular death signalling pathways. The c-Jun NH2-terminal kinases (JNKs) are classic stress-activated protein kinases and many cellular stresses have been shown to stimulate JNK activation. JNK in the pancreas is activated during brain death, pancreas procurement, and organ preservation, and its activity is progressively increased during the isolation procedure. Moreover, JNK activity in the transplanted liver after islet transplantation increases markedly within 24 hrs. Use of the JNK inhibitor in pancreas preservation, islet culture, and/or islet transplantation prevents islet apoptosis and improves islet graft function. These findings suggest that the control of JNK activation is important for pancreatic islet transplantation. PMID- 17124366 TI - Optimization of three-dimensional time-of-flight magnetic resonance angiography of the intracranial arteries. AB - The signal-to-noise ratio obtained from arteries in three-dimensional (3D) time of-flight (TOF) magnetic resonance (MR) angiography is often too low to allow clinical diagnosis because the radiofrequency pulse decreases the magnetization of protons in the blood and suppresses the in-flow effect in the slab. The present study adjusted the position of the head coil to boost arterial signal intensity. Ten healthy volunteers, eight men and two women aged 24-78 years, underwent 3D TOF MR angiography of the intracranial arteries with the same standard GE transmit-receive birdcage head coil using both normal and half position (lower edge of the coil level with the mouth) methods. Our subjects were divided into Group 1 consisted of five relatively young volunteers aged 24-42 years (mean 31.2 years), and Group 2 consisted of five older volunteers aged 70 78 years (mean 73 years). The following four arteries were chosen for analysis: the internal carotid artery (ICA), the proximal middle cerebral artery segment (M1), and the two distal middle cerebral artery segments (M2, M31). The half position method increased the signal-to-noise ratio in the ICA, M1, M2, and M3 by 15%, 25%, 36%, and 44%, respectively. In general, this method resulted in the generation of stronger signals in the M2 and M3 in younger subjects and in all arteries examined in older subjects. The half position method can provide better MR angiograms in certain brain regions of younger people, and in all brain regions in older patients. PMID- 17124367 TI - Incidence of ischemic lesions by diffusion-weighted imaging after carotid endarterectomy with routine shunt usage. AB - Temporary intraluminal shunt was used during 72 consecutive carotid endarterectomies (CEAs) in 61 patients (bilateral CEA in 11 patients) during October 2001 and September 2005. The medical records of these patients were retrospectively reviewed. All procedures were performed with routine shunt insertion without monitoring such as electroencephalography. Pre- and postoperative diffusion-weighted magnetic resonance (MR) imaging was used to detect ischemic complications. Postoperative angiography was performed in 70 cases to detect abnormalities such as major stenosis or dissection of the distal end. Symptomatic ischemic complication occurred in one patient at 1 month. Postoperative diffusion-weighted MR imaging detected new hyperintense lesions in three patients including the symptomatic patient. Postoperative angiography confirmed that the distal end was satisfactory in all cases. The incidence of ischemic lesions of embolic origin after CEA with routine shunt usage is acceptably low if the procedure of shunt device insertion and removal is meticulously conducted. PMID- 17124368 TI - Intra-arterial administration of fasudil hydrochloride for vasospasm following subarachnoid hemorrhage - analysis of time-density curve with digital subtraction angiography. AB - The cerebral circulatory dynamics were evaluated before and after intra-arterial administration of fasudil hydrochloride in 20 patients with angiographic vasospasm after subarachnoid hemorrhage (SAH). The region of interest time density curves obtained before and after intra-arterial administration of fasudil hydrochloride were compared in the proximal portion of the middle cerebral artery in the early arterial phase, the distal portion of the middle cerebral artery in the late arterial phase, and the transverse sinus in the venous phase. In the early arterial phase, the time to peak and the time to half-peak were significantly reduced. In the late arterial phase and venous phase, the time to peak was significantly reduced. These results suggest that intra-arterial administration of fasudil hydrochloride induced dilation of the proximal arteries, and improved cerebral microcirculation. The present study suggests that intra-arterial administration of fasudil hydrochloride is effective as a treatment for vasospasm following SAH. PMID- 17124369 TI - Hyperform remodeling balloon for the balloon occlusion test of persistent primitive trigeminal artery aneurysm - case report. AB - A 17-year-old woman presented with a rare aneurysm at the junction of the persistent primitive trigeminal artery (PPTA) and the internal carotid artery (ICA) manifesting as left abducens nerve paresis. The aneurysm and the ICA were both successfully occluded with coils. The balloon occlusion test used the HyperForm balloon microcatheter to seal the PPTA and ICA, which is very important to determine the optimal treatment strategy for a PPTA aneurysm. PMID- 17124370 TI - Endovascular treatment for a unusually large mycotic aneurysm manifesting as intracerebral hemorrhage - case report. AB - A 62-year-old male presented with an unusually large mycotic aneurysm mimicking a saccular aneurysm manifesting as coma and hypotension. Computed tomography showed intracerebral and intraventricular hemorrhage. He was in septic shock due to acute infectious endocarditis. Cerebral angiography disclosed a large distal anterior cerebral artery aneurysm. The diagnosis was mycotic aneurysm based on the morphological features and associated endocarditis. The aneurysm and the parent artery were successfully occluded by endovascular embolization. High-dose antibiotic therapy in the following 6 weeks resulted in resolution of the infectious endocarditis. Early exclusion of ruptured mycotic aneurysm is mandatory because of the high risk of rerupture. Endovascular treatment is an effective alternative for mycotic aneurysms, especially if the patient's general condition is poor. Parent artery occlusion can be safely tolerated if the aneurysm is located distally. PMID- 17124371 TI - Neuroendoscopic appearance of an intraventricular cavernous angioma blocking the foramen of monro - case report. AB - A 47-year-old woman presented with unilateral ventricular enlargement detected by magnetic resonance imaging during a medical checkup. Neuroendoscopic exploration identified a multilocular lesion in which dark red fluid formed a niveau near the right side of the foramen of Monro. The diagnosis was intraventricular cavernous angioma. Restricted flow of cerebrospinal fluid at the foramen of Monro was observed. Xanthochromia, which seemed to be due to previous bleeding, was observed at the fornix. When the neuroendoscope touched the angioma, the wall collapsed and bled. Endoscopic removal of the angioma was abandoned, and craniotomy and resection of the angioma were performed. No new neurological anomalies were observed after surgery. Preoperative diagnosis of intraventricular cavernous angioma is difficult based on neuroimaging. Neuroendoscopy is effective for diagnosis and the decision-making process regarding treatment. PMID- 17124372 TI - Intradural retroclival chordoma without bone involvement - case report. AB - A 63-year-old, previously healthy man presented with a rare large intradural retroclival chordoma without bone involvement. Computed tomography showed that the tumor was completely intradural and did not involve the bone, as confirmed at intraoperative inspection. The tumor was totally excised via the anterior transpetrosal approach. Surgery is the most effective first-line treatment for patients with chordoma despite the typical extradural extension and bone destruction. Complete resection is feasible for intradural extraosseous chordoma because of the sharply circumscribed margins and absence of bone involvement. Specialized skull base techniques should be used instead of conventional surgical approaches for intradural skull base chordoma. PMID- 17124373 TI - Ligamentum flavum hematoma in the cervical spine - case report. AB - A 67-year-old man presented with a rare case of ligamentum flavum hematoma manifesting as progressive tetraplegia following cervical traction therapy. Magnetic resonance imaging of the cervical spine showed a posterior mass that was continuous with the ligamentum flavum at the C3-4 levels. Complete resection of the mass that contained brownish hemorrhage was performed, resulting in excellent symptom relief. We speculate that repeated trivial trauma to the degenerative ligamentum flavum was the main predisposing factor in the present case. Ligamentum flavum hematoma is a rare cause of spinal root or cord compression which typically occurs in the lower thoracic or lumbar spine, but may also appear in the cervical spine. PMID- 17124374 TI - Congestive myelopathy due to cervical perimedullary arteriovenous fistula evaluated by apparent diffusion coefficient values - case report. AB - A 22-year-old woman presented with a cervical perimedullary arteriovenous fistula (AVF) manifesting as right upper and lower extremity weakness. T2-weighted magnetic resonance (MR) imaging showed intramedullary hyperintensity believed to be caused by venous congestion. Preoperative diffusion-weighted MR imaging showed increased apparent diffusion coefficient (ADC) value. Spinal angiography demonstrated an AVF fed mainly by the right C-5 radicular artery. Complete obliteration of AVF was achieved by endovascular embolization and microsurgical shunt occlusion. The ADC value was normalized and her neurological deficits improved after endovascular surgery, whereas T2-weighted MR imaging still demonstrated the lesion. The high preoperative ADC value probably indicated reversible vasogenic edema and immediate normalization of the ADC value suggests a good clinical outcome. PMID- 17124375 TI - Subcutaneous pneumocele associated with ventriculoperitoneal shunt migration into the mechanically occluded colon - case report. AB - A 62-year-old man presented with shunt failure manifesting as consciousness disturbance 4 years after placement of a ventriculoperitoneal shunt for subarachnoid hemorrhage. Physical examination found subcutaneous pneumocele around the peritoneal catheter extending from the abdomen to the neck. He had undergone pelvic radiation therapy for bladder cancer 2 years before. The peritoneal catheter was removed from the cervical region, and external ventricular drainage and a descending colon stoma for ileus release were positioned. The cerebrospinal fluid was clear and yielded no cultures. No inflammatory changes were seen. He developed carcinomatous peritonitis and died 4 months later. Retrograde colon gas reflux due to catheter perforation into the colon occluded by metastatic sigmoid cancer was probably the cause. Fragility of the wall of colon associated with the prior abdominal radiation therapy might have been a contributing factor. Subcutaneous pneumocele around the peritoneal catheter, i.e. pneumocele within the fibrous sheath surrounding the catheter, is a differential diagnosis to cerebrospinal fluid collection in patients with subcutaneous swelling around the catheter. PMID- 17124376 TI - The effects of hypoxia-ischemia on neutral amino acid transporters in the developing rat brain. AB - The neutral amino acid transporters SNAT1-3 and ASCT1 play critical roles in the recycling of glutamine, and subsequently glutamate, via the glutamine-glutamate cycle. Hypoxia-ischemia was induced in rat pups using the Rice-Vannucci model. Brains were harvested at 1 h, 24 h and 7 days after ischemia. The expression of NAATs was evaluated using immunoblotting, real-time PCR, and immunohistochemistry. Results were compared with age-matched controls and shams. SNAT1 mRNA decreased at 1 h after injury in both hemispheres when compared with the control animals and correlated with a decrease in protein expression at 24 h in the hippocampus and cortex. SNAT1 protein expression increased globally at 7 days after injury and specifically in the hippocampus. Finally, SNAT2 and 3 demonstrated subtle changes in various brain regions after injury. These data suggest that neutral amino acid transporters remain largely intact after hypoxia ischemia. PMID- 17124377 TI - Cross-talk between IGF-I and estradiol in the brain: focus on neuroprotection. AB - The actions of estradiol in the brain involve the interaction with growth factors, such as insulin-like growth factor-I (IGF-I). Many cells in the brain coexpress receptors for estradiol (ERs) and IGF-I (IGF-IR) and both factors interact to regulate neural function. Several studies have shown that there is an interaction of IGF-IR and ERs in neuroprotection. Neuroprotective effects of estradiol are blocked by the inhibition of IGF-IR signaling, while the neuroprotective effects of IGF-I are blocked by the inhibition of ER signaling. These findings suggest that the neuroprotective actions of estradiol and IGF-I after brain injury depend on the coactivation of both ERs and IGF-IR in neural cells. The relationship of ERalpha with IGF-IR through the phosphatidylinositol 3 kinase/Akt/glycogen synthase kinase 3beta (PI3K/Akt/GSK3) signaling pathway may represent the point of convergence used by estradiol and IGF-I to cooperatively promote neuroprotection. Administration of estradiol to ovariectomized rats results in the association of ERalpha with IGF-IR and with components of the PI3K/Akt/GSK3 signaling pathway and in the regulation of the activity of Akt and GSK3 in the brain. Conversely, IGF-I regulates ERalpha transcriptional activity in neuroblastoma cells and the PI3K/Akt/GSK3 signaling pathway is involved in this effect. PMID- 17124378 TI - Changes in the levels of interferon-gamma and transforming growth factor-beta influence bronchial stenosis during the treatment of endobronchial tuberculosis. AB - BACKGROUND: Endobronchial tuberculosis (EBTB) has been shown to frequently complicate bronchial stenosis, a condition which can induce dyspnea as a result of airway obstruction, and is also frequently misdiagnosed as either bronchial asthma or lung cancer. OBJECTIVES: This study attempted to determine whether there was a correlation between interferon-gamma (IFN-gamma) and transforming growth factor-beta (TGF-beta) levels in the serum and bronchial washing fluid (BWF), and the results of the treatment of EBTB patients. METHODS: Thirty patients, all of whom were diagnosed as EBTB, were enrolled, as were 10 healthy control subjects. IFN-gamma and TGF-beta levels were measured by the ELISA method in the serum and BWF of these 30 EBTB patients before and after treatment. The EBTB patients were divided into two groups: those who exhibited bronchial stenosis after treatment and those who did not. Chest computed tomography (CT) and pulmonary function test (PFT) were performed in 16 and 25 patients, respectively, at initial bronchoscopy. RESULTS: IFN-gamma and TGF-beta levels in the BWF of the EBTB patients were elevated compared to the controls (p < 0.05). After 2 months of treatment, 13 of the 30 EBTB patients exhibited bronchial fibrostenosis and the other 17 cases had recovered without sequelae. In the bronchial stenosis group, the initial serum TGF-beta levels were lower than in the patients without bronchial stenosis (p < 0.05). Moreover, the levels of serum TGF-beta after treatment were shown to have decreased more than in the patients without bronchial stenosis (p < 0.05). On chest CT findings of 16 EBTB patients, bronchial narrowing was suspected except in 2 cases (1 edematous-hyperemic type, 1 actively caseating type of segmental bronchus). The common features of PFT in EBTB at the initial diagnosis were a restrictive pattern and normal ventilatory function. CONCLUSIONS: Elevated IFN-gamma and TGF-beta levels in the BWF of the EBTB patients may be related to EBTB pathogenesis. Lowered initial serum TGF-beta levels as well as the observed changes in the levels of TGF-beta in the serum after treatment have been implicated in bronchial fibrostenosis during the course of the disease. PMID- 17124379 TI - Effect of fasting on cocaine-amphetamine-regulated transcript, neuropeptide Y, and leptin receptor expression in the non-human primate hypothalamus. AB - Leptin is a cytokine produced by white adipose tissue that circulates in direct proportion to adiposity and is an important signal of energy balance. Leptin inhibits food intake in rodents by inhibiting the orexigenic neuropetides neuropeptide Y (NPY) and agouti regulated peptide (AgRP) and stimulating the anorexigenic neuropeptides alpha-melanocyte-stimulating hormone (alpha-MSH) and cocaine-amphetamine-regulated transcript (CART). In order to extend our understanding of neuroendocrine regulation of appetite in the primate, we determined the effect of a metabolic challenge on CART, NPY, and leptin receptor (Ob-R) messenger ribonucleic acid (mRNA) in the nonhuman primate (NHP) hypothalamus. Ten adult female rhesus monkeys were either maintained on a regular diet or fasted for two days before euthanasia. CART, NPY, and Ob-R mRNA were measured by in situ hybridization histochemistry (ISHH). A 2-day fast decreased CART expression in the ARC, increased NPY gene expression in the supraoptic nucleus (SON) and paraventricular nucleus (PVN), and increased Ob-R expression in the ventromedial nucleus (VMN). This is the first report that fasting inhibits CART expression and stimulates Ob-R expression in monkeys. Increased NPY expression in the SON and PVN, but not the ARC of fasted monkeys also is novel. With some exceptions, our observations are confirmatory of findings in rodent studies. Similarities in the neuroendocrine responses to a metabolic challenge in monkeys and rodents support extending existing hypotheses of neuroendocrine control of energy homeostasis to primates. PMID- 17124380 TI - Absence of human herpesvirus 8 DNA sequences in lung biopsies from Israeli patients with pulmonary arterial hypertension. AB - BACKGROUND: Pulmonary hypertension is a severe pulmonary vascular disease leading to rapid deterioration and death. Histological and clinical evidence suggests that smooth muscle proliferation is part of the pathogenesis of the disease. Human herpesvirus 8 (HHV-8) is a gamma-herpesvirus that is implicated in malignancies and in Kaposi's sarcoma. Recently, the association of HHV-8 with idiopathic pulmonary arterial hypertension (PAH) has been found. OBJECTIVE: The aim of this study was to investigate the presence of HHV-8 in the lung tissue of Israeli patients with PAH. METHOD: The presence of HHV-8 sequences was investigated by polymerase chain reaction examination in 6 biopsies of patients with pulmonary hypertension. Three patients had idiopathic pulmonary hypertension, 2 patients pulmonary venoocclusive disease, and 1 patient pulmonary hypertension associated with mixed connective tissue disease. RESULT: We did not find any association between HHV-8 and PAH in these Israeli patients, as all the samples were negative for polymerase chain reaction. CONCLUSION: Our findings, together with the epidemiological data of HHV-8 prevalence and incidence rates of Kaposi's sarcoma and PAH in Israel, provide further evidence which argues against an association between HHV-8 infection and PAH. PMID- 17124381 TI - Efficacy and safety of the eXcelon transbronchial aspiration needle in mediastinal lymph node enlargement: a case-control study. AB - BACKGROUND: Several different types of needles for transbronchial needle aspiration (TBNA) exist in the market. Recently, the eXcelon transbronchial needle (Boston Scientific, Boston, Mass., USA) was commercialized, and to our knowledge, no comparative studies with other types of needles have been performed up to date. OBJECTIVES: The aim of the present study was to determine the diagnostic yield and safety of the 21-gauge eXcelon transbronchial needle in the diagnosis of mediastinal lesions and to compare it with a 22-gauge Wang cytology needle (Bard-Wang, Billerica, Mass., USA). METHODS: A case-control prospective study was conducted. The study group included 42 TBNA of mediastinal nodes performed in 35 patients with the eXcelon 21-gauge needle from January to June 2005. A control group included 110 TBNA carried out in 99 patients that underwent mediastinal lymph node sampling using a MW-122 needle from June 2002 to December 2004. RESULTS: There was no statistical differentiation in the characteristics of the disease, mediastinal lymph nodes or number of passes between cases and controls. Adequate samples were obtained in 30 (71.4%) cases and 81 (73.6%) controls (p = 0.8). Technical complications occurred in 7 (16.6%) cases and 14 (12.7%) controls (p = 0.7). The only clinical complication was bleeding presented in 2 (4.8%) cases and 3 (2.7%) controls (p = 0.3). CONCLUSION: The eXcelon 21 gauge transbronchial needle is as effective and safe as the MW-122 needle in the diagnosis of mediastinal lymph node enlargement. PMID- 17124382 TI - Intramuscular tumor detected by FDG positron emission tomography scanning following postoperative lung cancer. PMID- 17124383 TI - Blood pressure recordings within and outside the clinic and cardiovascular events in chronic kidney disease. AB - BACKGROUND: Blood pressure (BP) measured outside the clinic correlates better with cardiovascular outcomes in patients with essential hypertension. To assess the role of out-of-clinic BP recordings in predicting cardiovascular events in patients with chronic kidney disease (CKD), a prospective cohort study was conducted in 217 veterans with CKD. METHODS: BP was measured outside the clinic at home and by 24-hour ambulatory recordings, and in the clinic by 'routine' and standardized methods. Patients were followed over a median of 3.4 years to assess the combined end-point of total mortality, myocardial infarction or stroke. RESULTS: Average (+/-SD) home BP was 147.0 +/- 21.4/78.3 +/- 11.6 mm Hg, 24-hour ambulatory BP 133.5 +/- 16.6/73.1 +/- 11.1 mm Hg and in-clinic BPs were 155.2 +/- 25.6/84.7 +/- 14.2 mm Hg by the standardized method, and 144.5 +/- 24.2/75.4 +/- 14.7 mm Hg by the 'routine' method. A 1 SD increase in systolic BP increased the hazard ratio (HR) of the composite end-point by 1.16 (95% CI 0.89-1.50) for routine BP, 1.57 (95% CI 1.19-2.09) for standardized BP, 1.66 (95% CI 1.27-2.17) for home BP, and 1.42 (95% CI 1.10-1.84) for 24-hour ambulatory BP recording. The HR of the composite end-point was only significant for hypertension defined by 24 hour ambulatory BP monitoring (HR 2.22 (95% CI 1.23-4.01)). Adjusted for the propensity scores, BP measured by the ambulatory technique was not an independent predictor of cardiovascular events. Non-dipping was associated with increased cardiovascular risk, but not when adjusted for other risk factors. CONCLUSION: Risk factors that differentiate hypertension or non-dipping appear to confer a cardiovascular risk in CKD. PMID- 17124384 TI - Association of angiotensin I-converting enzyme gene insertion/deletion polymorphism and IgA nephropathy: a meta-analysis. AB - BACKGROUND/AIMS: The angiotensin I-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism has been extensively examined for the association with immunoglobulin A (IgA) nephropathy (IgAN), however, conflicting results have occurred. We performed a meta-analysis to evaluate the association of ACE I/D polymorphism with IgAN in different ethnic groups. METHODS: 11 studies testing the association between ACE I/D polymorphism and IgAN susceptibility, and 9 studies testing the association of ACE I/D with IgAN progression were used in this analysis. The overall odds ratio (OR) was estimated by a fixed or random effect model. RESULTS: The overall OR for the risk of susceptibility and progression of IgAN in Asians for the DD genotype is 2.37 (95% CI 1.04-5.41) and 1.75 (95% CI 1.24-2.56). The overall OR for the D allele in Asians also showed a similar magnitude, though without statistical significance (p = 0.09, p = 0.13, respectively). In Caucasians, both the DD genotype and D allele were associated with IgAN progression (OR 1.90, 1.61, respectively), but not IgAN susceptibility (p = 0.30, p = 0.41, respectively). CONCLUSION: Our findings support the notion that ACE I/D polymorphism is associated with IgAN. Meanwhile, the role of ACE I/D polymorphism in Asians is different from that of Caucasians. PMID- 17124385 TI - HGF protects rat mesangial cells from high-glucose-mediated oxidative stress. AB - BACKGROUND: Oxidative stress has been considered to be a common pathogenetic factor of diabetic nephropathy. Recent observations suggested that hepatocyte growth factor (HGF) was an antioxidant growth factor; thus, its renoprotective effects in diabetic nephropathy might be related to antioxidant mechanism. The aim of the present study was to evaluate whether HGF could prevent rat mesangial cells (RMC) from high-glucose-mediated oxidative stress and explore its relevant mechanism. METHODS: RMC were cultured in 5.6 mM (NG) or 30 mM (HG) glucose in the absence or presence of HGF (20 ng/ml) and c-met inhibitor SU11274 (5 microM) for 24 h. RESULTS: c-met expression in HG was markedly increased. Enhanced oxidative stress was observed in HG as evidenced by elevated reactive oxygen species and malondialdehyde levels and decreased glutathione level, which was markedly attenuated by HGF. HGF also inhibited HG-induced p22(phox) and aldose reductase upregulation and prevented HG-reduced glutamate-cysteine ligase catalytic subunit (GCLC) expression through inhibiting USF binding to negative regulatory region of GCLC promoter. Reduced glucose-6-phosphate dehydrogenase activity and expression in RMC by HG was rescued by HGF. CONCLUSION: HGF could function as an antioxidant factor and protect against HG-mediated oxidative stress by enhancing ROS scavenging and suppressing ROS production. PMID- 17124386 TI - Cosegregation of a novel homozygous CYP11B1 mutation with the phenotype of non classical congenital adrenal hyperplasia in a consanguineous family. AB - We report a novel missense mutation of CYP11B1 causing non-classical 11beta hydroxylase deficiency in 3 members of a consanguineous Turkish family. Two siblings presented with clinical evidence of precocious pseudopubarche. Biochemistry suggested 11beta-hydroxylase deficiency and genetic analysis revealed that they were homozygous for the missense mutation L489S within exon 9 of the CYP11B1 gene. The unaffected parents were heterozygotes for the same mutation. In addition, a paternal aunt of the affected siblings presenting with primary infertility and mild hirsutism was found to have the same homozygous mutation. This is the first report of a homozygous mutation in non-classical congenital adrenal hyperplasia that cosegregates with clinical phenotype. The significance of the missense mutation L489S in CYP11B1 is further supported by the conservation of leucine at position 489 in CYP11 genes in eleven other species. Molecular modelling of the enzyme suggests that the mutation L489S in CYP11B1 may alter the enzyme's substrate-binding affinity. These findings suggest that this homozygous mutation affects 11beta-hydroxylase function, resulting in the clinical features of non-classical adrenal hyperplasia in this family. PMID- 17124387 TI - Symptomatic intracranial hemorrhage after thrombolysis. PMID- 17124388 TI - Multiparametric iterative self-organizing data analysis of ischemic lesions using pre- or post-Gd T1 MRI. AB - BACKGROUND: The purpose of this work was to evaluate effects of Gd diethylenetriaminepentacetic acid (DTPA) injection on T(1)-weighted images of stroke and lesion segmentation and characterization results generated by our multiparametric iterative self-organizing data (ISODATA) method. The post-Gd image incorporates vasculature information into the analysis. METHODS: Either a pre-Gd T(1)-weighted image (T1WI) or a post-Gd T1WI was used along with diffusion , T(2)- and proton-density-weighted images in the analysis. ISODATA is a data driven method that segments and characterizes tissue damage in stroke using multiparametric MRI. RESULTS: Experimental results in both animal and human studies showed that the use of post-Gd T1WI modified the segmentation and characterization results on the periphery of the lesion. The peripheral region that changes with Gd-DTPA has a higher permeability compared to the rest of the lesion. Either of the data sets (including pre- or post-Gd T1WI) was used to estimate the tissue recovery and generated consistent results. CONCLUSIONS: This study shows that our multiparametric ISODATA approach consistently identifies and characterizes the core of the ischemic lesion. It also shows that the inclusion of post-Gd T1WI results in the segmentation and characterization of the lesion periphery if it has a higher permeability compared to the rest of the lesion. Finally, it confirms that the multiparametric ISODATA MRI characterizes tissue damage and recovery in stroke. PMID- 17124389 TI - Fluoxetine is not effective in the treatment of post-stroke fatigue: a double blind, placebo-controlled study. AB - BACKGROUND AND PURPOSE: Although post-stroke fatigue (PoSF) is common, pharmacological interventions to improve PoSF have rarely been carried out. The purpose of the present study was to evaluate the therapeutic effect of fluoxetine on PoSF. METHODS: We studied 83 consecutive outpatients with PoSF at an average of 14 months after the onset of stroke. The presence of post-stroke depression, post-stroke emotional incontinence and post-stroke anger proneness was also evaluated with the use of a standardized questionnaire. The presence of PoSF and pre-stroke fatigue was assessed. The visual analogue scale (VAS) and Fatigue Severity Score (FSS) were used to assess PoSF. The subjects were given either 20 mg/day of fluoxetine (n = 40) or placebo (n = 43) for 3 months. Follow-up evaluations were done 3 and 6 months after the beginning of the treatment. RESULTS: The initial mean fatigue VAS score and the mean overall FSS score were 5.4 +/- 2.0 and 4.4 +/- 1.2, respectively. There were no differences in the number of patients with PoSF between the fluoxetine group and the placebo group at 3 and 6 months after the treatment. The percent changes in VAS scores and FSS at all follow-up assessments were not significantly different either. However, fluoxetine significantly improved post-stroke emotional incontinence (p < 0.05) and post-stroke depression (p = 0.05) in the patients with PoSF. CONCLUSIONS: Fluoxetine does not improve PoSF, although some concomitant emotional disturbances improved significantly. Our results suggest that PoSF may be associated with diverse etiologies but not closely related to serotonergic dysfunction. Further studies are required to elucidate the causative factors and to find an appropriate treatment for PoSF. PMID- 17124390 TI - Antithrombotic therapy and predilection for cerebellar hemorrhage. AB - BACKGROUND: With the recent increase in the use of antithrombotic therapy, intracerebral hemorrhage (ICH) has been found to be a common complication. We determined whether the use of oral antithrombotic therapy and the patients' preexisting comorbidities were predictive of cerebellar hemorrhage (CH; previously reported to be associated with anticoagulants) as compared to other ICH, and whether antithrombotic therapy affected the clinical severity of CH. METHODS: A study of 327 consecutive patients hospitalized in our institute within 3 days after the onset of ICH, including 38 patients with a CH. RESULTS: CH accounted for 12% of all ICH, 75% of which occurred in patients on warfarin therapy with an international normalized ratio (INR) for prothrombin time >2.5 (p < 0.0001), and 33% of which occurred in patients on ticlopidine therapy (p = 0.017). Warfarin therapy with an INR >2.5 and high blood glucose on admission were independently predictive of CH as compared to other ICH. In addition, previous ischemic stroke (p = 0.002) and heart diseases (p = 0.018) were more prevalent in patients with CH than in those with other ICH. The number of major arteriosclerotic comorbidities and risk factors was also independently predictive of CH risk. CONCLUSIONS: We confirmed that warfarin therapy with an INR >2.5 is associated with CH. Patients with CH frequently had arteriosclerotic comorbidities requiring antithrombotic therapy that can complicate their acute management. PMID- 17124391 TI - Recurrence after ischemic stroke in chinese patients: impact of uncontrolled modifiable risk factors. AB - BACKGROUND: Data concerning stroke occurrence and recurrence in China are extremely rare. This study was designed to analyze determinants of stroke recurrence in a cohort of Chinese patients. METHODS: Subjects were patients with ischemic stroke registered in the Nanjing Stroke Registry Program. Modifiable risk factors for stroke were identified and stratified into 3 levels: without, controlled and uncontrolled. Cox proportional hazard model was used to detect influence factors for stroke recurrence. RESULTS: First-year recurrence rate was 11.2% in the registered patients. Hypertension, atrial fibrillation (AF) and smoking were associated with increased risk of recurrence. Controlling hypertension and AF each halved recurrent risk (p < 0.001). Ceasing smoking for more than 1 year reduced hazard ratio of recurrence from 1.71 to 1.39 (p < 0.05). Controlling blood sugar level in diabetics did not significantly change recurrent risk (hazard ratio, 1.69 vs. 1.64, p > 0.05). CONCLUSIONS: The recurrence rate is higher in Chinese patients with ischemic stroke compared with the one reported in western populations. Failure to control some modifiable risk factors in Chinese patients may be responsible for this discrepancy. PMID- 17124392 TI - The ruler is dead: an analysis of carotid plaque motion. AB - Factors involved in atherosclerotic plaque progression are still a topic of hot debate. With the advance of new ultrasound techniques it has been possible to identify atherosclerotic plaque characteristics that may reflect a higher risk of vulnerability. Among the various proposed mechanisms, one of the least investigated is plaque motion, i.e. plaque stability in respect to the arterial wall. We report here two asymptomatic patients with internal carotid stenosis that showed differences in plaque motion, a finding that could determine a change in their future treatment. PMID- 17124393 TI - Trends in the incidence of transient ischemic attacks, premorbid risk factors and the use of preventive treatments in the population of Dijon, France from 1985 to 2004. AB - BACKGROUND: We describe the epidemiological trends of transient ischemic attack (TIA) in a 20-year population-based pilot study. METHODS: Trends in the incidence, risk factors and pre-TIA use of preventive treatments for TIA were observed from 1985 to 2004 according to the classic definition in the population of the city of Dijon, France (150,000 inhabitants). RESULTS: The raw and standardized incidence of TIA were stable over time. We observed a significant increase in the mean age at TIA onset in women only. The prevalence of hypercholesterolemia and diastolic blood pressure > or =90 mm Hg among patients with TIA increased significantly. This contrasts with falls in smoking and in history of previous myocardial infarction. CONCLUSION: The stability of classic TIA incidence, despite the rise in the proportion of elderly people, and the increase in the mean age at onset in women may be considered as a medical progress. PMID- 17124394 TI - The Khorasan Stroke Registry: results of a five-year hospital-based study. AB - The hospital-based stroke registry is useful for understanding the diverse clinical characteristics of stroke related to geographical, racial or environmental differences. The Khorasan Stroke Registry (KSR) was established for the evaluation of incidence, clinical manifestations, risk factors, topography and etiology of ischemic stroke in Southern Khorasan, Iran, during 2001-2005. Consecutive stroke patients underwent a standard battery of diagnostic investigations by a stroke neurologist. The topography and etiology of brain infarction were determined by the Practical Iranian Criteria classification. The incidence of ischemic stroke in the Persian population is 43.17 cases per 100,000 people per year. A total of 1,392 ischemic stroke patients (738 females, 654 males) were evaluated in the KSR. The etiologies included atherosclerosis (53.6%), followed by uncertain causes (19.9%), cardioembolism (11.8%) and miscellaneous etiologies (2.9%). Of our patients, 11.7% had both atherosclerosis and cardioembolic mechanisms. Rheumatic valvular disease was present in 44.8% of cardioembolic stroke patients and caused 4.31 preventable stroke cases per 100,000 Persian people per year. Hypertension and history of ischemic cerebrovascular events were the most frequent risk factors, 53.1 and 22.3%, respectively. The in-hospital mortality of our ischemic stroke patients was 7.3%. A total of 336 patients with intracerebral hemorrhage (189 females, 147 males) were evaluated in the KSR. The incidence of intracerebral hemorrhage in the Persian population is 10.43 cases per 100,000 people per year. The high frequency of atherosclerotic etiology in the KSR is due to its classification criteria, which do not separate small vessel territory infarcts as a different etiologic subtype. Rheumatic valvular disease is an important cause of stroke in the Persian population. PMID- 17124395 TI - Microcirculatory dysfunction in the brain precedes changes in evoked potentials in endotoxin-induced sepsis syndrome in rats. AB - BACKGROUND: During sepsis progression microcirculatory dysfunction precedes macrocirculatory failure, partly explaining the occurrence of early organ dysfunction. The matter concerning microcirculatory dysfunction in the brain under septic conditions is less clear. We investigated the integrity of the activation flow coupling during sepsis progression in a rat model of septic shock. METHODS: Chloralose-anesthetized rats (n = 30) were subjected to electric forepaw stimulation. Over the somatosensory cortex electrical activity and hemodynamic responses were recorded with surface electrodes and laser Doppler. After baseline recordings, vehicle, 1 or 5 mg/kg lipopolysaccharide (LPS) from Escherichia coli was given intravenously, and activation flow coupling, blood pressure and blood gases were investigated at regular time points up to 270 min. In the end lactate, glucose, neuron-specific enolase (NSE) and S-100B protein levels were measured. RESULTS: Besides stable data from the control group, all LPS-treated rats developed signs of septic shock, which were more pronounced in the 5 mg/kg LPS group. Cerebral hyperemia occurred and was similar between the sepsis groups despite lower blood pressure levels in the 5 mg/kg LPS group. While the activation flow coupling remained intact in the 1 mg/kg LPS group, an uncoupling occurred in the 5 mg/kg group. First, the evoked flow velocity responses dropped 60 min after sepsis induction before the somatosensory amplitudes also decreased 120 min later. From similar NSE levels we suggest a functional rather than structural deficit explaining the difference in evoked potentials. CONCLUSIONS: For the first time we demonstrate microcirculatory dysfunction in the activation flow coupling of the brain. Inappropriate blood supply of neurons might explain the disturbance of neuronal function. PMID- 17124396 TI - Polymorphisms in genes of the renin-angiotensin system and cerebral small vessel disease. AB - BACKGROUND: Genetic variation in the renin-angiotensin system (RAS) has been implicated in stroke, particularly the small vessel disease (SVD) subtype. Furthermore, there may be two distinct subtypes of cerebral SVD, isolated lacunar infarction (ILI) and ischaemic leukoaraiosis (ILA). METHODS: 300 patients with well-phenotyped SVD and 600 controls were genotyped for five polymorphisms in the angiotensinogen (AGT) gene and eight polymorphisms within the angiotensin converting enzyme (ACE) gene. RESULTS: AGT and ACE polymorphisms and haplotypes were no more common in SVD cases as a whole or the two subtypes. Amongst hypertensives only, an AGT promoter polymorphism (-20A-->C), was associated with the ILA subtype (multivariate odds ratio 1.716, 95% confidence interval 1.073 2.746, p = 0.024). CONCLUSIONS: RAS genetic variants are not strong risk factors for cerebral SVD. The AGT -20C allele may be a risk factor for the leukoaraiosis subtype amongst hypertensives. PMID- 17124397 TI - How good is the management of vascular risk after stroke, transient ischaemic attack or carotid endarterectomy? AB - BACKGROUND: Patients remain at high risk of vascular events after stroke, transient ischaemic attack or carotid endarterectomy. We studied how well this risk is addressed by the effective treatment of modifiable risk factors. METHODS: A total of 198 consecutive attenders at a rapid access stroke clinic and 98 consecutive patients undergoing carotid endarterectomy were studied. Treatment of hypertension and hyperlipidaemia, smoking status and the use of antithrombotic therapy were assessed at baseline and 6 months later. The findings were compared with targets from the UK National Clinical Guidelines for Stroke. RESULTS: Baseline and follow-up data were available on 284 patients. The rates of control of vascular risk factors improved only slightly during follow-up. Blood pressure was below target levels in only 69 (24%) at baseline and 79 (28%) at 6 months, and serum cholesterol was below target levels in only 55 (19%) at baseline and 63 (22%) at 6 months. At baseline, 55 (19%) were smokers, of whom 12 (22%) had quit at 6 months. Anticoagulant therapy was prescribed in 19 of 37 patients (51%) in atrial fibrillation at 6 months. Antiplatelet therapy was prescribed in 90% of patients in sinus rhythm. CONCLUSIONS: Despite the identification of vascular risk factors at the time of clinic or surgery, 6 months later these risk factors remain poorly addressed. More effective methods of managing vascular risk in these patients are needed. PMID- 17124398 TI - Improved survival after stroke: is admission to hospital the major explanation? Trend analyses of the Auckland Regional Community Stroke Studies. AB - BACKGROUND: There is uncertainty regarding the impact of changes in stroke care and natural history of stroke in the community. We examined factors responsible for trends in survival after stroke in a series of population-based studies. METHODS: We used statistical models to assess temporal trends in 28-day and 1 year case fatality after first-ever stroke cases registered in 3 stroke incidence studies undertaken in Auckland, New Zealand, over uniform 12-month calendar periods in 1981-1982 (n = 1,030), 1991-1992 (1,305) and 2001-2002 (1,423). Cox proportional hazards regression was used to evaluate the significance of pre defined 'patient', 'disease' and 'service/care' factors on these trends. RESULTS: Overall, there was a 40% decline in 28-day case fatality after stroke over the study periods, from 32% (95% confidence interval, 29-35%) in 1981-1982 to 23% (21 25%) in 1991-1992 and then 19% (17-21%) in 2002-2003. Similar relative declines were seen in 1-year case fatality. In regression models, the trends were still significant after adjusting for patient and disease factors. However, further adjustment for care factors (higher hospital admission and neuroimaging) explained most of the improvement in survival. CONCLUSIONS: These data show significant downwards trends in case fatality after stroke in Auckland over 20 years, which can largely be attributed to improved stroke care associated with increases in hospital admission and brain imaging during the acute phase of the illness. PMID- 17124399 TI - Impact of clinical environment on embolus detection: a comparison of automated and manual detection of Doppler embolic signals. AB - BACKGROUND: Transcranial Doppler ultrasound detection of weak embolic signals is inhibited by intrinsic limitations within the human auditory system. Psychoacoustics effects are likely to be exacerbated in a clinical environment, where automated embolus detection has potential to surpass manual detection. In this study we quantify the impact of clinical environment on manual detection of Doppler embolic signals following carotid surgery. We also discuss the implications of psychoacoustics considerations for the evaluation of automated detection systems. METHOD: Concurrent monitoring by vascular technologists and an automated embolus detection system were performed for 50 consecutive patients during postoperative recovery. Both detection methods were evaluated against a majority decision human expert panel analyzing under ideal conditions. RESULTS: Clinical environment reduced the overall sensitivity of manual monitoring by approximately 23%, mainly due to a approximately 2-dB increase in the lower threshold for detection. Clinical environment was also associated with a reduction in positive predictive value for manual detection of approximately 9% compared to ideal conditions. Automated monitoring, which is not affected by environment, was marginally more sensitive for detection of weaker embolic signals. CONCLUSIONS: One in 4 weak embolic signals was missed during routine clinical monitoring compared to ideal conditions. Automated detection (in this study) performed slightly better than human observers but did not approach the performance of the majority decision panel. PMID- 17124400 TI - Spontaneous recanalization of occluded standard extracranial-intracranial arterial bypass. AB - BACKGROUND: In the event of early failure of standard extracranial-intracranial arterial bypass, elective but prompt revision surgery is generally attempted under the assumption that early occlusion is permanent. However, little is known about the occurrence of spontaneous revascularization. OBJECTIVE: To estimate the frequency and time course of spontaneous recanalization in primary extracranial intracranial arterial bypass occlusion and re-evaluate diagnostic and therapeutic options facing spontaneous development. METHODS AND RESULTS: In a retrospective analysis 176 patients with standard superficial temporal artery/middle cerebral artery bypass were included. Twenty-three patients had primary bypass occlusion. In 7 cases spontaneous recanalization was observed. Follow-up of these cases is presented. CONCLUSIONS: Early bypass occlusion has a high incidence of spontaneous recanalization within the first year. In stable patients the first choice may be noninvasive follow-up and postponing revision operation. PMID- 17124401 TI - Historical development of analysing large-scale changes in the human genome. AB - A widely held belief today is that genomics really only started with the DNA sequence information emanating from the genome programs for various organisms, with the human genome playing the leading role. In fact there is a discernable trail stretching for more than a 100 years from the observations of Boveri on tissue instability involving polyploidy in sea urchin embryos and human tumours to the present day. This historical review follows that trail and shows that many theoretical and technical advantages taken for granted in today's genomics era rely heavily on earlier cytogenetic and gene mapping discoveries. Three specific examples of technical developmental paths involving in situ hybridisation, flow sorting and DNA reassociation kinetics will be explored. In the mid-1980s the two former approaches merged to give rise to several applications of which chromosome painting and chromosome CGH are arguably the most important. The latter developed into array CGH which has now become the pre-eminent method for detecting micro imbalances in a large number of targets. A competing emerging technology is that of genome-wide SNP typing, which itself is a product of the much earlier RFLP approach linked to DNA sequence information. Do such approaches spell the final demise of the microscope? Perhaps for narrowly defined activities this may occur, but for addressing general questions, microscopic examination will remain pre eminent. PMID- 17124402 TI - Development of bioinformatics resources for display and analysis of copy number and other structural variants in the human genome. AB - The discovery of an abundance of copy number variants (CNVs; gains and losses of DNA sequences >1 kb) and other structural variants in the human genome is influencing the way research and diagnostic analyses are being designed and interpreted. As such, comprehensive databases with the most relevant information will be critical to fully understand the results and have impact in a diverse range of disciplines ranging from molecular biology to clinical genetics. Here, we describe the development of bioinformatics resources to facilitate these studies. The Database of Genomic Variants (http://projects.tcag.ca/variation/) is a comprehensive catalogue of structural variation in the human genome. The database currently contains 1,267 regions reported to contain copy number variation or inversions in apparently healthy human cases. We describe the current contents of the database and how it can serve as a resource for interpretation of array comparative genomic hybridization (array CGH) and other DNA copy imbalance data. We also present the structure of the database, which was built using a new data modeling methodology termed Cross-Referenced Tables (XRT). This is a generic and easy-to-use platform, which is strong in handling textual data and complex relationships. Web-based presentation tools have been built allowing publication of XRT data to the web immediately along with rapid sharing of files with other databases and genome browsers. We also describe a novel tool named eFISH (electronic fluorescence in situ hybridization) (http://projects.tcag.ca/efish/), a BLAST-based program that was developed to facilitate the choice of appropriate clones for FISH and CGH experiments, as well as interpretation of results in which genomic DNA probes are used in hybridization-based experiments. PMID- 17124403 TI - Idiopathic learning disability and genome imbalance. AB - Learning disability (LD) is a very common, lifelong and disabling condition, affecting about 3% of the population. Despite this, it is only over the past 10 15 years that major progress has been made towards understanding the origins of LD. In particular, genetics driven advances in technology have led to the unequivocal demonstration of the importance of genome imbalance in the aetiology of idiopathic LD (ILD). In this review we provide an overview of these advances, discussing technologies such as multi-telomere FISH and array CGH that have already emerged as well as new approaches that show diagnostic potential for the future. The advances to date have highlighted new considerations such as copy number polymorphisms (CNPs) that can complicate the interpretation of genome imbalance and its relevance to ILD. More importantly though, they have provided a remarkable approximately 15-20% improvement in diagnostic capability as well as facilitating genotype/phenotype correlations and providing new avenues for the identification and understanding of genes involved in neurocognitive function. PMID- 17124404 TI - Molecular karyotyping of patients with MCA/MR: the blurred boundary between normal and pathogenic variation. AB - Molecular karyotyping has revealed that microdeletions/duplications in the human genome are a major cause of multiple congenital anomalies associated with mental retardation (MCA/MR). The identification of a de novo chromosomal imbalance in a patient with MCA/MR is usually considered causal for the phenotype while a chromosomal imbalance inherited from a phenotypically normal parent is considered as a benign variation and not related to the disorder. Around 40% of imbalances in patients with MCA/MR in this series is inherited from a healthy parent and the majority of these appear to be (extremely) rare variants. As some of these contain known disease-causing genes and have also been found to be de novo in MCA/MR patients, this challenges the general view that such familial variants are innocent and of no major phenotypic consequence. Rather, we argue, that human genomes can be tolerant of genomic copy number variations depending on the genetic and environmental background and that different mechanisms play a role in determining whether these chromosomal imbalances manifest themselves. PMID- 17124405 TI - Cytogenetic genotype-phenotype studies: improving genotyping, phenotyping and data storage. AB - High-resolution molecular cytogenetic techniques such as genomic array CGH and MLPA detect submicroscopic chromosome aberrations in patients with unexplained mental retardation. These techniques rapidly change the practice of cytogenetic testing. Additionally, these techniques may improve genotype-phenotype studies of patients with microscopically visible chromosome aberrations, such as Wolf Hirschhorn syndrome, 18q deletion syndrome and 1p36 deletion syndrome. In order to make the most of high-resolution karyotyping, a similar accuracy of phenotyping is needed to allow researchers and clinicians to make optimal use of the recent advances. International agreements on phenotype nomenclature and the use of computerized 3D face surface models are examples of such improvements in the practice of phenotyping patients with chromosomal anomalies. The combination of high-resolution cytogenetic techniques, a comprehensive, systematic system for phenotyping and optimal data storage will facilitate advances in genotype phenotype studies and a further deconstruction of chromosomal syndromes. As a result, critical regions or single genes can be determined to be responsible for specific features and malformations. PMID- 17124406 TI - Copy number variation in the genome; the human DMD gene as an example. AB - Recent developments have yielded new technologies that have greatly simplified the detection of deletions and duplications, i.e., copy number variants (CNVs). These technologies can be used to screen for CNVs in and around specific genomic regions, as well as genome-wide. Several genome-wide studies have demonstrated that CNV in the human genome is widespread and may include millions of nucleotides. One of the questions that emerge is which sequences, structures and/or processes are involved in their generation. Using as an example the human DMD gene, mutations in which cause Duchenne and Becker muscular dystrophy, we review the current data, determine the deletion and duplication profile across the gene and summarize the information that has been collected regarding their origin. In addition we discuss the methods most frequently used for their detection, in particular MAPH and MLPA. PMID- 17124407 TI - Impact of low copy repeats on the generation of balanced and unbalanced chromosomal aberrations in mental retardation. AB - Low copy repeats (LCRs) are stretches of duplicated DNA that are more than 1 kb in size and share a sequence similarity that exceeds 90%. Non-allelic homologous recombination (NAHR) between highly similar LCRs has been implicated in numerous genomic disorders. This study aimed at defining the impact of LCRs on the generation of balanced and unbalanced chromosomal rearrangements in mentally retarded patients. A cohort of 22 patients, preselected for the presence of submicroscopic imbalances, was analysed using submegabase resolution tiling path array CGH and the results were compared with a set of 41 patients with balanced translocations and breakpoints that were mapped to the BAC level by FISH. Our data indicate an accumulation of LCRs at breakpoints of both balanced and unbalanced rearrangements. LCRs with high sequence similarity in both breakpoint regions, suggesting NAHR as the most likely cause of rearrangement, were observed in 6/22 patients with chromosomal imbalances, but not in any of the balanced translocation cases studied. In case of chromosomal imbalances, the likelihood of NAHR seems to be inversely related to the size of the aberration. Our data also suggest the presence of additional mechanisms coinciding with or dependent on the presence of LCRs that may induce an increased instability at these chromosomal sites. PMID- 17124408 TI - Whole-genome array-CGH screening in undiagnosed syndromic patients: old syndromes revisited and new alterations. AB - We report array-CGH screening of 95 syndromic patients with normal G-banded karyotypes and at least one of the following features: mental retardation, heart defects, deafness, obesity, craniofacial dysmorphisms or urogenital tract malformations. Chromosome imbalances not previously detected in normal controls were found in 30 patients (31%) and at least 16 of them (17%) seem to be causally related to the abnormal phenotypes. Eight of the causative imbalances had not been described previously and pointed to new chromosome regions and candidate genes for specific phenotypes, including a connective tissue disease locus on 2p16.3, another for obesity on 7q22.1-->q22.3, and a candidate gene for the 3q29 deletion syndrome manifestations. The other causative alterations had already been associated with well-defined phenotypes including Sotos syndrome, and the 1p36 and 22q11.21 microdeletion syndromes. However, the clinical features of these latter patients were either not typical or specific enough to allow diagnosis before detection of chromosome imbalances. For instance, three patients with overlapping deletions in 22q11.21 were ascertained through entirely different clinical features, i.e., heart defect, utero-vaginal aplasia, and mental retardation associated with psychotic disease. Our results demonstrate that ascertainment through whole-genome screening of syndromic patients by array CGH leads not only to the description of new syndromes, but also to the recognition of a broader spectrum of features for already described syndromes. Furthermore, on the technical side, we have significantly reduced the amount of reagents used and costs involved in the array-CGH protocol, without evident reduction in efficiency, bringing the method more within reach of centers with limited budgets. PMID- 17124409 TI - Array-based comparative genomic hybridization and copy number variation in cancer research. AB - Array-based comparative genomic hybridization (aCGH) is a molecular cytogenetic technique used in detecting and mapping DNA copy number alterations. aCGH is able to interrogate the entire genome at a previously unattainable, high resolution and has directly led to the recent appreciation of a novel class of genomic variation: copy number variation (CNV) in mammalian genomes. All forms of DNA variation/polymorphism are important for studying the basis of phenotypic diversity among individuals. CNV research is still at its infancy, requiring careful collation and annotation of accumulating CNV data that will undoubtedly be useful for accurate interpretation of genomic imbalances identified during cancer research. PMID- 17124410 TI - Genome wide measurement of DNA copy number changes in neuroblastoma: dissecting amplicons and mapping losses, gains and breakpoints. AB - In the past few years high throughput methods for assessment of DNA copy number alterations have witnessed rapid progress. Both 'in house' developed BAC, cDNA, oligonucleotide and commercial arrays are now available and widely applied in the study of the human genome, particularly in the context of disease. Cancer cells are known to exhibit DNA losses, gains and amplifications affecting tumor suppressor genes and proto-oncogenes. Moreover, these patterns of genomic imbalances may be associated with particular tumor types or subtypes and may have prognostic value. Here we summarize recent array CGH findings in neuroblastoma, a pediatric tumor of the sympathetic nervous system. A total of 176 primary tumors and 53 cell lines have been analyzed on different platforms. Through these studies the genomic content and boundaries of deletions, gains and amplifications were characterized with unprecedented accuracy. Furthermore, in conjunction with cytogenetic findings, array CGH allows the mapping of breakpoints of unbalanced translocations at a very high resolution. PMID- 17124411 TI - Gene copy number changes in dermatofibrosarcoma protuberans - a fine-resolution study using array comparative genomic hybridization. AB - Dermatofibrosarcoma protuberans (DFSP) is a rare, slow-growing, low-grade dermal tumor. Cytogenetic and FISH studies have revealed that the chromosomal rearrangements characteristic of DFSP tumors involve both translocations and the formation of a supernumerary ring derived from chromosomes 17 and 22. The t(17;22) (q22;q13.1) translocation generates a gene fusion between COL1A1 and PDGFB, which serves as a diagnostic marker of DFSP. In the present study we performed array-CGH (aCGH) analysis on ten DFSP tumors. The COL1A1 region at 17q was gained in 71% (5/7) of the samples and the PDGFB region at 22q was gained in 43% (3/7) of the individual samples. In addition to the 17q and 22q gains, altogether 17 minimal common regions of gain and one region of loss were detected. PMID- 17124412 TI - Molecular parameters associated with insulinoma progression: chromosomal instability versus p53 and CK19 status. AB - Insulinomas represent the predominant syndromic subtype of endocrine pancreatic tumors (EPTs). Their metastatic potential cannot be predicted reliably using histopathological criteria. In the past few years, several attempts have been made to identify prognostic markers, among them TP53 mutations and immunostaining of p53 and recently cytokeratin 19 (CK19). In a previous study using conventional comparative genomic hybridization (CGH) we have shown that chromosomal instability (CIN) is associated with metastatic disease in insulinomas. It was our aim to evaluate these potential parameters in a single study. For the determination of CIN, we applied CGH to microarrays because it allows a high resolution detection of DNA copy number changes in comparison with conventional CGH as well as the analysis of chromosomal regions close to the centromeres and telomeres, and at 1pter-->p32, 16p, 19 and 22. These regions are usually excluded from conventional CGH analysis, because they may show DNA gains in negative control hybridizations. Array CGH analysis of 30 insulinomas (15 tumors of benign, eight tumors of uncertain and seven tumors of malignant behavior) revealed that >or=20 chromosomal alterations and >or=6 telomeric losses were the best predictors of malignant progression. A subset of 22 insulinomas was further investigated for TP53 exon 5-8 gene mutations, and p53 and CK19 expression. Only one malignant tumor was shown to harbor an arginine 273 serine mutation and immunopositivity for p53. CK19 immunopositivity was detected in three malignant tumors and one tumor with uncertain behavior. In conclusion, our results indicate that CIN as well as telomeric loss are very powerful indicators for malignant progression in sporadic insulinomas. Our data do not support a critical role for p53 and CK19 as molecular parameters for this purpose. PMID- 17124413 TI - Lung cancer genetics and pharmacogenomics. AB - Epidermal growth factor receptor (EGFR)-targeted therapies have demonstrated remarkable success in a small subset of non-small cell lung cancer patients. The mechanism of response has been an area of active research, with somatic mutation in a number of genes in the EGFR signal transduction pathway and copy number alterations of genes of the EGFR family as candidates contributing towards response. Continuing studies should help determine an appropriate biomarker or combination of biomarkers that can be used to predict response to this class of therapy. PMID- 17124414 TI - Medical applications of array CGH and the transformation of clinical cytogenetics. AB - Microarray-based comparative genomic hybridization (array CGH) merges molecular diagnostics with traditional chromosome analysis and is transforming the field of cytogenetics. Prospective studies of individuals with developmental delay and dysmorphic features have demonstrated that array CGH has the ability to detect any genomic imbalance including deletions, duplications, aneuploidies and amplifications. Detection rates for chromosome abnormalities with array CGH range from 5-17% in individuals with normal results from prior routine cytogenetic testing. In addition, copy number variants (CNVs) were identified in all studies. These CNVs may include large-scale variation and can confound the diagnostic interpretations. Although cytogeneticists will require additional training and laboratories must become appropriately equipped, array CGH holds the promise of being the initial diagnostic tool in the identification of visible and submicroscopic chromosome abnormalities in mental retardation and other developmental disabilities. PMID- 17124415 TI - Stability of neurocognitive impairment in different subtypes of mild cognitive impairment. AB - There has been increasing interest in delineating different cognitive subtypes of mild cognitive impairment (MCI). It remains unclear, however, the extent to which neuropsychological impairment associated with amnestic, nonamnestic, and amnestic+ subtypes of MCI remains stable over time. In this study, 70 persons meeting the criteria for MCI and 38 cognitively normal elderly subjects received a baseline neuropsychological evaluation and were reevaluated 1 year later. Our results indicated that 84.6% of the persons initially classified as amnestic, 75% of those classified as nonamnestic, and 80% of the persons classified as MCI+ evidenced stable or more pronounced neuropsychological impairment across the follow-up period. Less than 7% of the amnestic and amnestic+ cases had nonimpaired neuropsychological profiles at their reevaluation at 12 months, and 16.7% of the nonamnestic cases had nonimpaired neuropsychological test profiles at follow-up. Approximately 87% of the cognitively normal subjects at baseline continued to have unimpaired neuropsychological performance at follow-up. These results indicate that the presence of neuropsychological impairment is relatively stable over a 12-month follow-up period among different cognitive subtypes of MCI, although 15-25% of the cases did not exhibit the specific cognitive deficits that characterized their performance at baseline. PMID- 17124416 TI - Differential CSF biomarker levels in APOE-epsilon4-positive and -negative patients with memory impairment. AB - OBJECTIVES: To investigate the relationships between episodic memory, APOE genotype, CSF markers (total tau, T-tau; phospho-tau, P-tau; beta-amyloid, Abeta42) and longitudinal cognitive decline. METHODS: 124 memory clinic patients were retrospectively divided into 6 groups based on (i) episodic memory function (Rey Auditory Verbal Learning Test, RAVLT): severe, moderate or no impairment (SIM, MIM or NIM), and (ii) APOE genotype (epsilon4+ or epsilon4-). CSF marker levels and cognitive decline were compared across groups. RESULTS: Episodic memory function, according to RAVLT scores, was significantly correlated with CSF marker levels only among epsilon4+ subjects and not among epsilon4- subjects. When comparing the 6 subgroups, SIM epsilon4+ and MIM epsilon4+ groups showed significantly lower Abeta42 levels than the other groups. T-tau and P-tau levels were significantly increased in SIM epsilon4+ when compared to all the other groups, including the SIM epsilon4- group. However, both SIM epsilon4+ and SIM epsilon4- declined cognitively during the follow-up. CONCLUSION: It remains to be determined whether APOE genotype affects the expression of biomarkers in CSF, or whether the different biomarker patterns reflect different types of disease processes in patients with progressive cognitive dysfunction. PMID- 17124417 TI - Pyomyoma in a perimenopausal woman with intrauterine device. AB - Pyomyoma (suppurative leiomyoma) is a rare disease, which is considered to be a serious complication of uterine leiomyoma. Since 1945, only 18 patients have been reported and ours is the 19th. Although it is frequently reported in pregnant women or postmenopausal women who have vascular disease, our case is a 42-year old woman in the perimenopausal period who presented with fever and a tender lower abdominal mass. She used the intrauterine device as a contraceptive method but leiomyoma had never been found before. Ultrasonographic findings suggested an ovarian tumor. She was diagnosed as having infected malignant ovarian cancer with an elevated CA 125 level that was initially treated with broad spectrum antibiotics; then she underwent total abdominal hysterectomy and bilateral salpingo-oophorectomy. Pathological findings showed acute and chronic inflammation of the endometrium with abscess formation in an intramural leiomyoma. The intrauterine device might be the origin of pyomyoma due to a direct spread of the infection from the uterine cavity. Pyomyoma may be difficult to diagnose especially in those with a nonspecific clinical presentation without any history of leiomyoma. Delayed diagnosis may result in serious complications, whereas adequate surgery and broad spectrum antibiotics may decrease serious morbidity and mortality. PMID- 17124418 TI - Sex and age specificity of susceptibility genes modulating survival at old age. AB - OBJECTIVE: We aimed to investigate the influence of the genetic variability of candidate genes on survival at old age in good health. METHODS: First, on the basis of a synthetic survival curve constructed using historic mortality data taken from the Italian population from 1890 onward, we defined three age classes ranging from 18 to 106 years. Second, we assembled a multinomial logistic regression model to evaluate the effect of dichotomous variables (genotypes) on the probability to be assigned to a specific category (age class). Third, we applied the regression model to a cross-sectional dataset (10 genes; 972 subjects selected for healthy status) categorized according to age and sex. RESULTS: We found that genetic factors influence survival at advanced age in good health in a sex- and age-specific way. Furthermore, we found that genetic variability plays a stronger role in males than in females and that, in both genders, its impact is especially important at very old ages. CONCLUSIONS: The analyses presented here underline the age-specific effect of the gene network in modulating survival at advanced age in good health. PMID- 17124419 TI - Can we expect something from prohepcidin measurement in hemodialysis patients? PMID- 17124421 TI - Is bcl-2 a key molecule in the abnormal immune response of hemodialysis patients? PMID- 17124422 TI - Effect of the hemodialysis session on bcl-2 expression in peripheral blood mononuclear cells in vivo. AB - bcl-2 is a proto-oncogene with a regulatory role in many conditions due to its marked inhibitory action on apoptosis. Reports regarding the effect of hemodialysis (HD) on apoptosis of mononuclear cells and in association with bcl-2 expression in particular, are controversial. The aim of the present study was to examine in vivo the influence of an HD session on bcl-2 expression of lymphocytes and monocytes. We measured quantitative bcl-2 expression with flow cytometry, in terms of antibodies bound per cell, in blood samples taken from 44 HD patients before and after an HD session. 27 patients (group I) were dialyzed with synthetic-type membranes and 17 (group II) with cellulose-type membranes. bcl-2 expression increased statistically significantly in lymphocytes (1,616 +/- 718 to 1,894 +/- 715 molecules/cell, p < 0.01) at the end of HD. Monocyte expression of bcl-2 was lower than in lymphocytes and almost did not change after the HD session (654 +/- 446 to 698 +/- 375 molecules/cell, p = NS). Comparison between the two groups did not reveal a significant difference in either the baseline bcl 2 expression or in the value of the increase after HD. We conclude that HD seems to decrease lymphocyte apoptosis independent of the biocompatibility of the dialyzer membrane. PMID- 17124423 TI - Daptomycin clearance during modeled continuous renal replacement therapy. AB - BACKGROUND/AIMS: Pharmacotherapy in critically ill patients receiving continuous renal replacement therapies (CRRT) is challenging due to the lack of published information to base dosing regimens. METHODS: Daptomycin's transmembrane clearance during continuous hemofiltration and hemodialysis was assessed using an in vitro model with AN69 and polysulfone hemodiafilters at varying ultrafiltrate and dialysate flow rates (1, 2, 3 and 6 l/h). RESULTS: During continuous hemofiltration, mean daptomycin sieving coefficient ranged from 0.14 to 0.20. Transmembrane clearances were significantly different between filter types for ultrafiltration rates of 2, 3 and 6 l/h. For continuous hemodialysis, mean daptomycin saturation coefficient ranged from 0.05 to 0.15. AN69-based daptomycin clearances were significantly lower than polysulfone values at dialysate flow rates of 2, 3 and 6 l/h. CONCLUSION: The extent of daptomycin's transmembrane clearance is dependent on hemodiafilter type, dialysate and ultrafiltration rates. CRRT with high ultrafiltrate or dialysate rates may result in substantial daptomycin clearances. PMID- 17124424 TI - A new semiempirical mathematical model for prediction of internal filtration in hollow fiber hemodialyzers. AB - The potential of convective solute transport for blood purification has been widely explored. New techniques (such as hemodiafiltration), based on a combination of diffusion and convection techniques, have been developed. Owing to the internal filtration/backfiltration (IF/BF) phenomenon, high-flux dialysis also relies on a convective component, which, however, is hard to quantify and thus optimize. In this work, we developed a mathematical model designed to supply the clinician with a quantification of the IF/BF fluxes taking place during high flux dialysis. IF fluxes are predicted based on the machine settings and blood hematocrit/protein concentration. The hydraulic characteristics of commercial dialyzers were derived from bloodless bench tests. Moreover, an in vitro blood test was conducted on a 1.8 m(2) polysulfone dialyzer using an established scintigraphic analysis, for verification of model prediction accuracy. Results of simulations show that the IF/BF rate is sensitive to the blood flow rate and (to a lesser extent) to the dialysate flow rate. Increasing net ultrafiltration rates resulted in parallel increases of direct filtration and simultaneous decreases of BF. IF/BF is rather influenced by blood composition, due to the complex dependence of oncotic pressure and blood viscosity upon hematocrit and plasma protein concentration. Simulation results showed an excellent agreement with the experimental results obtained with scintigraphy, with only a 3% prediction error. With respect to some previous works, this model is simpler in its theoretical approach. It allows implementation into a user-friendly software tool and might be used to predict the convective component in high-flux dialysis and possibly to optimize it. PMID- 17124425 TI - Dialysis-related genotoxicity: sister chromatid exchanges and DNA lesions in T and B lymphocytes of uremic patients. Genomic damage in patients on hemodiafiltration. AB - BACKGROUND/AIMS: Patients with chronic renal failure show the presence of massive oxidative genome damage but the role played by dialysis is still a controversial issue. The aim of our study was to verify the genomic damage in B- and T lymphocyte subpopulations of uremic patients after a single hemodiafiltration session. METHODS: We enrolled 30 patients on maintenance acetate-free biofiltration and 25 age-matched healthy volunteers and studied chromosomal alterations. RESULTS: Our data show that the basal levels of DNA damage, the number of sister chromatid exchanges and basal high-frequency cells levels are significantly higher in patients on hemodiafiltration than in controls and in T lymphocytes than in B cells. CONCLUSIONS: These findings suggest that hemodialytic treatment could represent a potential source of damage, maybe through the oxidative action of the extracorporeal circuit components, which might explain the well-known T-specific immunodeficiency correlated with uremia. PMID- 17124426 TI - Peroxisome proliferator-activated receptor-gamma agonists diminish peritoneal functional and morphological changes induced by bioincompatible peritoneal dialysis solution. AB - BACKGROUND: To evaluate if peroxisome proliferator-activated receptor-gamma (PPAR gamma) agonists have a potential protective effect on the peritoneum changes induced by bioincompatible peritoneal dialysis (PD) solution in vivo. METHODS: Male Wistar rats were dialyzed three times daily for 28 days with 1.36% Dianeal (two groups: with (D+R) or without (D) rosiglitazone) or 1.36% Physioneal (two groups: with (P+R) or without (P) rosiglitazone). Peritoneal transport of fluid and small solutes was assessed. Nine rats that did not receive dialysis served as controls. RESULTS: Significant morphological changes were found in the D group compared with controls. Additional use of rosiglitazone in the D+R group resulted in less morphological changes and expression of collagen I as well as an increased drainage volume. The expression of VEGF was inhibited by rosiglitazone while no apparent effect was found regarding TGF/Smad pathway. CONCLUSIONS: The addition of rosiglitazone to standard dialysis fluids can maintain the peritoneal morphology and increase ultrafiltration in a PD rat model. PMID- 17124428 TI - Role of cytokines in allergic airway inflammation. AB - Asthma is characterized by intense infiltration of eosinophils and CD4+ T cells into the submucosal tissue of airways. Accumulating evidence indicates that T helper type 2 cell-derived cytokines such as interleukin (IL)-4, IL-5 and IL-13 play critical roles in orchestrating and amplifying allergic inflammation in asthma. In addition, it has been suggested that newly identified cytokines including thymic stromal lymphopoietin, IL-25 and IL-33 are involved in the induction of allergic inflammation in asthma. In this review, we discuss the role of individual cytokines in the pathogenesis of asthma. PMID- 17124429 TI - A hybrid expressing genetically engineered major allergens of the Parietaria pollen as a tool for specific allergy vaccination. AB - BACKGROUND: Allergy is an immunological disorder affecting about 25% of the population living in the industrialized countries. Specific immunotherapy is the only treatment with a long-lasting relief of allergic symptoms and able to reduce the risk of developing new allergic sensitizations and inhibiting the development of clinical asthma in children treated for allergic rhinitis. METHODS: By means of DNA recombinant technology, we were able to design a head to tail dimer expressing disulphide bond variants of the major allergen of the Parietaria pollen. IgE binding activity was studied by Western blot, ELISA inhibition assays and the skin prick test. T cell recognition was studied by peripheral blood mononuclear cell proliferation. The immunogenicity of the hybrid was studied in a mouse model of sensitization. RESULTS: In vitro and in vivo analysis showed that the disruption of specific cysteine residues in both allergens caused a strong reduction in IgE binding activity of the PjEDcys hybrid. In addition,we were able to show that a reduction in the IgE epitope content profoundly reduced the anaphylactic activity of the hybrid (from 100 to 1,000 times less than wild-type allergens) without interfering with the T cell recognition. Sera from BALB/c mice immunized with the hybrid were able to bind the natural Parietaria allergens and to inhibit the binding of human IgE to wild-type Par j 1 and Par j 2 allergens up to 90%. CONCLUSION: Our results demonstrate that hybrid-expressing disulphide bond variants of the major allergens of the Parietaria pollen displayed reduced allergenicity and maintained T cell reactivity for induction of protective antibodies. PMID- 17124430 TI - Elevated soluble ADAM8 in bronchoalveolar lavage fluid in patients with eosinophilic pneumonia. AB - BACKGROUND: ADAM (a disintegrin and metalloprotease) family members, characterized by a metalloprotease and a disintegrin domain, are membrane anchored glycoproteins involved in proteolysis and cell adhesion. ADAM8 might have an important role in allergic inflammation. It can cleave a variety of substrates and is a sheddase for VCAM-1 and CD23, the low-affinity IgE receptors. METHODS: To evaluate the contribution of ADAM8 to the pathogenesis of eosinophilic pneumonia (EP), we measured the concentrations of soluble ADAM8 (sADAM8) and its substrates, soluble VCAM-1 (sVCAM-1) and soluble CD23 (sCD23), in bronchoalveolar lavage fluid from patients with smoking-induced acute eosinophilic pneumonia (AEP), chronic idiopathic eosinophilic pneumonia (CEP), and drug-induced eosinophilic pneumonia (drug-EP). RESULTS: The sADAM8 and sVCAM 1 concentrations were increased in AEP and CEP. The sCD23 concentration was elevated in AEP. In AEP, but not CEP, the sADAM8 concentration significantly correlated with those of both sVCAM and sCD23. CONCLUSION: The pathogenesis of AEP, CEP, and drug-EP was distinct with regard to ADAM8. Our results are the first to associate ADAM8 with eosinophilic responses and lung inflammation in humans. PMID- 17124431 TI - Effects of pravastatin treatment on blood pressure regulation after renal transplantation. AB - BACKGROUND/AIM: Hypertension is one of the main cardiovascular risk factors and has an impact also on long-term kidney graft survival. In addition to their lipid lowering properties, it was shown that 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors also have a blood pressure lowering effect. We examined whether treatment with a statin interferes with blood pressure regulation and antihypertensive treatment after renal transplantation. METHODS: 74 patients were treated with initially 20 mg of pravastatin daily immediately after kidney transplantation. This group was compared to a matched cohort of 76 patients without statin treatment. All patients received standard immunosuppressive triple therapy with ciclosporin A microemulsion together with an antiproliferative agent and prednisolone. Primary objective of this analysis was systolic and diastolic blood pressure regulation with and without pravastatin. Furthermore, graft function expressed as creatinine clearance and proteinuria, immunosuppressive regimen, and incidence of cardiovascular events and graft loss were recorded for 48 months. RESULTS: The blood pressure regulation was comparable in both groups; however, to achieve this, significantly more antihypertensive drugs had to be used in the statin-treated patients as compared with the controls (2.9 vs. 2.2 agents at 48 months). A slightly higher ciclosporin A exposure of the statin treated patients could have contributed to this observation. The graft function after 4 years was comparable between the groups (creatinine clearance 56.9 vs. 57.0 ml/min), and a trend of reduced proteinuria could be demonstrated after 4 years of statin treatment (0.4 vs. 0.9 g/day). The low-density lipoprotein cholesterol levels decreased as expected during treatment (3.1 vs. 3.7 mmol/l at 48 months), but the recommended target levels for patients with a high cardiovascular risk have not been reached. A trend towards lower incidences of acute rejection, chronic allograft nephropathy, and graft loss was noted in the statin-treated group. Adverse effects of statin treatment have not been observed. CONCLUSION: Treatment with pravastatin at low to average dosages does not result in improved blood pressure regulation after kidney transplantation. PMID- 17124432 TI - Norepinephrine transporter gene (NET) polymorphism in patients with type 2 diabetes. AB - BACKGROUND: Norepinephrine transporter (NET) is involved in the regulation of norepinephrine (NE) turnover and metabolism. Neuronal NE reuptake may be impaired in individuals with renal disease and/or hypertension due to dysfunction of the NE transporter. A silent G1287A nucleotide substitution in exon 9 of the NET gene was studied in human conditions involving hypertension. We investigated its effect in patients with type 2 diabetes. METHODS: The study involved 215 type 2 diabetes patients with nephropathy, 95 patients with diabetes duration > or =10 years, free of nephropathy, and 360 healthy subjects. All individuals were genotyped for the NET-8 gene polymorphism with the PCR-RFLP method. Genotype and allele frequencies were compared between the groups. NE was measured by high performance liquid chromatography and electrochemical detection. RESULTS: We genotyped 310 patients and 360 controls for the NET gene polymorphism. Genotype distribution in both groups was in accordance with the Hardy-Weinberg equilibrium. There were no significant differences in the frequency of genotypes and alleles between patients and controls (p = 0.43). The frequencies were also similar for patients with nephropathy and those without. After dividing the patient group into hypertensive (n = 208) and normotensive (n = 102) subjects, there was a significant increase in the frequency of the AA genotype in patients with hypertension compared to normotensives (19 vs. 10%, p < 0.05). CONCLUSION: No association was found between G1287A polymorphism in the NET gene and diabetes. Our results suggest that this polymorphism has a possible role in increased susceptibility to hypertension in patients with type 2 diabetes. PMID- 17124433 TI - A downloadable three-dimensional virtual model of the visible ear. AB - PURPOSE: To develop a three-dimensional (3-D) virtual model of a human temporal bone and surrounding structures. METHODS: A fresh-frozen human temporal bone was serially sectioned and digital images of the surface of the tissue block were recorded (the 'Visible Ear'). The image stack was resampled at a final resolution of 50 x 50 x 50/100 micro m/voxel, registered in custom software and segmented in PhotoShop 7.0. The segmented image layers were imported into Amira 3.1 to generate smooth polygonal surface models. RESULTS: The 3-D virtual model presents the structures of the middle, inner and outer ears in their surgically relevant surroundings. It is packaged within a cross-platform freeware, which allows for full rotation, visibility and transparency control, as well as the ability to slice the 3-D model open at any section. The appropriate raw image can be superimposed on the cleavage plane. The model can be downloaded at: (https://research.meei.harvard.edu/Otopathology/3dmodels/). PMID- 17124434 TI - Clinicopathological features of patients with concomitant intraductal papillary mucinous neoplasm of the pancreas and pancreatic endocrine neoplasm. AB - BACKGROUND/AIMS: The occurrence of concomitant pancreatic endocrine neoplasm (PEN) and intraductal papillary neoplasm (IPMN) of the pancreas has rarely been reported. We describe our experience with 3 patients with this association and review the existing literature. METHODS: From 1990 to 2005, 65 patients who underwent surgery for a PEN or IPMN were retrospectively reviewed. Forty-three patients had a PEN, 19 had an IPMN and 3 had both an IPMN and PEN. The 3 patients with concomitant IPMN and PEN are the focus of the current study and their clinicopathological features are reported together with 7 patients previously reported in the literature. RESULTS: There were 10 patients with a median age of 62 years (range 40-73). The male:female ratio was equal. Seven of 10 patients were symptomatic and the most common symptoms were abdominal pain (n = 5), jaundice (n = 2) and loss of weight (n = 2). The median size of the endocrine neoplasms was 14 mm (range 2-30) and they occurred in the head (n = 3), body (n = 2) and tail (n = 5). Seven of the PENs were classified as benign, 2 were potentially malignant, and 1 was frankly malignant with lymph node involvement. None of the endocrine neoplasms were functioning. The IPMNs were found in the tail (n = 4), head (n = 3), head and body (n = 1), body (n = 1) and the entire pancreas (n = 1). Five of these neoplasms were benign, 2 were borderline and 3 were malignant (1 carcinoma in situ). CONCLUSION: The occurrence of concomitant IPMN and PEN is more frequent than would be expected. However, it is difficult in the present analysis to determine if this association is more than just fortuitous. PMID- 17124435 TI - Endoscopic retrograde cholangiopancreatography-induced severe acute pancreatitis. AB - BACKGROUND/AIMS: There is scant information in the literature about the outcomes of endoscopic retrograde cholangiopancreatography (ERCP)-induced severe acute pancreatitis (ESAP). Compared to other causes, higher morbidity and mortality have been reported in ERCP-induced acute pancreatitis. We undertook this study to determine the differences between ESAP and SAP due to other causes (OSAP). METHODS: We retrospectively identified all cases of SAP admitted to our institution during the years 1992-2001. We reviewed the medical records of all SAP patients to obtain information on demographics, interventions, local and systemic complications and outcomes. RESULTS: We identified 207 patients with SAP, of whom 16 (7.7%) had ESAP and 191 OSAP. There was no difference between ESAP and OSAP with regard to demographics, clinical interventions, local and systemic complications and outcomes. Both groups had a similar mortality (25 vs. 18%). CONCLUSION: ESAP has a similar morbidity and mortality compared to OSAP. PMID- 17124436 TI - Polymorphism in the IL-8 gene, but not in the TLR4 gene, increases the severity of acute pancreatitis. AB - BACKGROUND/AIM: Activated granulocytes and inflammatory mediators of the innate immune response play fundamental roles in the pathogenesis of acute pancreatitis. We studied whether polymorphisms of interleukin-8 (IL-8) and Toll-like receptor 4 (TLR4) genes correlate with the severity of acute pancreatitis. METHODS: Patients with acute pancreatitis (n = 92) were grouped according to the severity of the disease on the basis of the Ranson scores. Healthy blood donors (n = 200) served as controls. The IL-8 -251 gene polymorphism was analyzed by amplification refractory mutation system; the single-nucleotide polymorphisms (Asp299Gly and Thr399Ile) of TLR4 were investigated by using a real-time polymerase chain reaction method with melting point analysis. RESULTS: The IL-8 A/T heterozygote mutant variants were detected with a significantly higher frequency among the patients with severe pancreatitis than among the healthy blood donors (60 vs. 42%; p = 0.0264, odds ratio = 2.071, 95% confidence interval = 1.101-3.896), while the frequency of the normal allelic genotype (TT) was higher among the patients with mild pancreatitis than in the group with severe pancreatitis (35 vs. 16%; p = 0.051, odds ratio = 2.917, 95% confidence interval = 1.089-7.811). There was no significant correlation between TLR4 polymorphisms and the acute pancreatitis itself, but nonsignificantly increased frequencies of Asp299Gly and Thr399Ile heterozygotes among patients with severe infected pancreatic necrosis could be observed relative to the patients with mild pancreatitis. CONCLUSIONS: Determination of the frequency of IL-8 polymorphism in acute pancreatitis may be informative and may provide further evidence concerning the role of IL-8 in the severe form of this disease. The possible role of TLR4 polymorphism in the outcome of severe acute pancreatitis requires further investigations in a larger series of patients. PMID- 17124437 TI - [Intratympanic treatment in Meniere's disease: the effect of gentamicin and dexamethasone on vertigo control and hearing]. AB - OBJECTIVES: We investigated the effect of intratympanic gentamicin and dexamethasone treatments on the control of vertigo and hearing loss in patients with Meniere's disease. PATIENTS AND METHODS: Forty-five patients with a diagnosis of Meniere's disease according to the 1995 criteria of American Academy of Otorhinolaryngology Head and Neck Surgery were assigned to receive intratympanic gentamicin (40 mg/ml, 0.7 ml; n=24; 11 males, 13 females; mean age 46 years) or dexamethasone (4 mg/ml, 0.7 ml; n=21; 8 males, 13 females; mean age 50.4 years). The results were evaluated with respect to changes in vertigo and hearing symptoms. RESULTS: In the gentamicin group, vertigo symptoms were controlled in 22 patients (92%), deterioration in hearing was seen in only two patients (8%). In the dexamethasone group, nine patients had complete follow-up. Of these, vertigo control was achieved in six patients (67%), none had worsened hearing, and one patient (5%) had improved hearing. When improvement in hearing was defined as at least a 5 dB change, then five patients (24%) benefited from treatment. CONCLUSION: Intratympanic treatment modalities in Meniere's disease are easy to perform, cheap, and effective. They are expected to have a more extensive use with higher success rates in the future. PMID- 17124438 TI - Comparison of two endoscopic examination methods, the Muller maneuver and fiberoptic pharyngoscopy during sleep, in patients with obstructive sleep apnea. AB - OBJECTIVES: This study was designed to compare two endoscopic examination methods, the Muller maneuver (MM) and fiberoptic pharyngoscopy during sleep, both of which are used to detect obstructed segments in patients with obstructive sleep apnea. PATIENTS AND METHODS: The study included 28 patients (23 males, 5 females; mean age 44.6 years; range 28 to 59 years) who underwent uvulopalatopharyngoplasty (UPPP) for snoring or obstructive sleep apnea. Obstruction was examined both at the level of the soft palate and tongue base while the patients were awake and asleep and was scored. The Muller maneuver was performed in the sitting and supine positions. In addition, fiberoptic pharyngoscopy was performed right after induction of anesthesia. The results of the two methods were compared. RESULTS: Changes in body position were not associated with significant differences in the results of MM. The two methods were found to be highly discordant, in that a greater degree of obstruction was noted especially at the level of the soft palate by fiberoptic pharyngoscopy. CONCLUSION: It was concluded that the degree of obstruction might be underestimated by MM. PMID- 17124439 TI - [Ear symptoms accompanying temporomandibular joint diseases]. AB - OBJECTIVES: We investigated symptoms of otologic diseases in patients with temporomandibular joint diseases. PATIENTS AND METHODS: The study included 44 patients (26 females, 18 males; mean age 36+/-10.7 years; range 21 to 58 years) with temporomandibular joint disease. A questionnaire was administered to the patients to inquire into otological symptoms accompanying temporomandibular joint disease. RESULTS: Temporomandibular joint disease was bilateral in 11 patients. Accompanying otological symptoms were as follows: click voice with jaw movements (n=21; 47.7%), headache (n=15; 34.1%), pain in the ear with jaw movements (n=14; 31.8%), tinnitus (n=8; 18.2%), fullness in the ear (n=6; 13.6%), vertigo or dizziness (n=6; 13.6%), and hearing loss (n=3; 6.8%). Pure-tone audiometry revealed sensorineural hearing loss in one patient, being 28 dB on the right and 30 dB on the left. CONCLUSION: Our findings suggest that otologic symptoms may accompany temporomandibular joint diseases, requiring that patients presenting with otologic symptoms must be investigated for temporomandibular joint diseases. PMID- 17124440 TI - [The effect of topical application of 5-fluorouracil on patency times after incisional myringotomy of guinea pig tympanic membrane]. AB - OBJECTIVES: We investigated the effect of topical application of 5-fluorouracil after incisional myringotomy on the patency time and its use as an alternative to ventilation tube insertion. MATERIALS AND METHODS: Experimental tympanic membrane perforations were induced in two groups of guinea pigs. One group (20 tympanic membranes) was treated with a 50 mg/ml 5-fluorouracil-soaked sponge over the perforation for five minutes while the other (16 tympanic membranes) with saline soaked sponges for the same duration. The patency times, otorrhea, and other complications were recorded. RESULTS: The mean patency of tympanic membrane perforations following incisional myringotomy was significantly longer in the 5 fluorouracil-treated group (11.3 days vs 7.0 days; p<0.01). None of the ears exhibited permanent perforation. CONCLUSION: The length of patency provided by 5 fluorouracil is not sufficient for optimal ventilation time in otitis media with effusion. However, in circumstances in which shorter patency times may provide sufficient ventilation, 5-fluorouracil application may be an alternative to ventilation tube insertion. PMID- 17124441 TI - The effect of systemic prednisolone on propylene glycol-induced otitis media in guinea pig. AB - OBJECTIVES: In this histopathologic study, we investigated the effect of systemic prednisolone on propylene glycol-induced otitis media in guinea pig ear. MATERIALS AND METHODS: Ten adult guinea pigs received systemic prednisolone (10 mg, IM, single dose) together with a single application of 60% propylene glycol (0.2 ml, per ear) to both middle ears. Ten control guinea pigs were given a single application of 60% propylene glycol (0.2 ml) in the left ear, and a single application of saline (0.2 ml) in the right ear. Four weeks later, the animals were sacrificed and temporal bones were removed and prepared for histological evaluation. Paraffin-embedded specimens were horizontally sectioned at 7 microns. One of each five sections was stained with hematoxylin and eosin, and studied under light microscopy. RESULTS: In the control group, otitis media developed in six of nine propylene glycol-administered ears (66%), whereas all saline-treated ears remained intact. In the prednisolone group, only one (6%) of 16 ears developed otitis media and cholesteatoma. The difference was statistically significant. CONCLUSION: Our results suggest that systemic prednisolone has a potent effect to prevent propylene glycol-induced otitis media in guinea pig ears. PMID- 17124442 TI - An unusual foreign body: a ball bearing in the posterior ethmoid sinus. AB - Globe perforation was detected in an 18-year-old male patient having total vision loss in the left eye following a car accident. The middle meatus could not be evaluated by sinonasal endoscopy because of edema. Computed tomography showed a foreign body in the left posterior ethmoid sinus, with a trajectory coursing from the globe to the lamina papyracea, and the posterior ethmoid sinus. Endoscopic sinus surgery was performed including a left uncinectomy, anterior and posterior ethmoidectomies, and the foreign body was removed from the posterior ethmoid sinus using ethmoid forceps. It turned out to be a ball bearing. After three months of follow-up, the patient had no evidence for rhinologic sequelae. PMID- 17124443 TI - [Tracheal adenoid cystic carcinoma presenting as a thyroid tumor]. AB - Primary tumors of the trachea are rare. Adenoid cystic carcinoma is the second most common tracheal malignancy and its symptoms are nonspecific. A 66-year-old woman presented with complaints of progressive dyspnea and cough of a two-week duration. She had received asthmatic treatment many times for the past four years. Physical examination revealed a large, firm, nontender, multilobular mass in the thyroid gland. Indirect laryngoscopic examination showed subglottic stenosis. On a plain cervical radiograph, there was narrowing of the tracheal lumen. Emergent tracheotomy was performed and a biopsy was taken from the mass. Histopathologic examination revealed adenoid cystic carcinoma, suggesting a thyroid mass with a tracheal origin. Despite medical treatment, the patient died in the fourth month. PMID- 17124444 TI - [Childhood peripheral vestibular disorders: a report of three cases]. AB - Vertigo is uncommon in childhood. Its etiology is different from that of adults. Both central and peripheral disorders may give rise to vertigo symptoms in pediatric patients. The evaluation of vertigo in children requires a detailed history taking, clinical examination, audiometric assessment, and vestibular function tests. We present three pediatric patients with vertigo symptoms, together with treatment methods and results of treatment. Their ages ranged from 10 to 14 years. The diagnoses were vestibular neuritis in one patient, and benign paroxismal positional vertigo in two patients. Treatment was comprised of vestibular rehabilitation for vestibular neuritis, and canalith repositioning maneuver for benign paroxismal positional vertigo. All the patients benefited well from the procedures applied. PMID- 17124445 TI - [Tracheobronchial foreign body aspiration following partial laryngectomy: a case report]. AB - Tracheobronchial foreign body aspiration is uncommon in adults and one etiologic factor is laryngeal surgery. A 50-year-old male patient presented with cough, high fever, and severe dyspnea three months after partial laryngectomy for squamous cell carcinoma of the larynx. A computed tomography scan of the thorax showed a hyperdense mass lesion in the right main bronchus, suggesting a foreign body. Diagnostic bronchoscopy was performed under general anesthesia and a small bone fragment, 2 x 1.3 cm in size, was removed from the right bronchial lumen. The patient showed slow improvement due to prolonged atelectasis until complete clinical and radiologic recovery. PMID- 17124446 TI - [A general review on the procedures in a human temporal bone laboratory]. AB - Structures of the human ear are usually inaccessible during life for examination of pathologic changes of the underlying disease, which is only possible with postmortem studies of the human temporal bone. Human temporal bone laboratories serve as a unique source of material for research in this respect. They enable comparison between histologic findings of temporal bone sections and the ear pathologies documented prior to death, as well as comparison of diseased ears with any selected temporal bone specimens, both of which provide invaluable knowledge to be shared among researchers and other laboratories. This article aims to provide insight into the functions of temporal bone laboratories and to familiarize the reader with histopathologic studies conducted therein. PMID- 17124447 TI - [Diagnosis and treatment of soft-tissue tumors]. AB - The diagnostic and therapeutic management of patients with soft-tissue tumors would be similar to the approach used for bone tumors if it were not for one crucial factor: the absolute necessity to recognize a sarcoma. The predominant features are the size of the tumor and its superficial or deep localization. If the tumor is small and superficial, biopsy can be associated with immediate resection without risk of dissemination to the deep tissues: this is the biopsy resection approach. If the tumor is deep or superficial but large sized, search for locoregional spread with MRI is necessary before undertaking any surgical procedure. MRI can help guide the biopsy and plan resection if the tumor is a sarcoma. A first biopsy is necessary to establish the histological diagnosis and elaborate the therapeutic strategy. Samples should be sent immediately to the pathology lab which should examine sterile fresh tissue. Experience has demonstrated that proper rules for diagnosis and treatment are not necessarily applied initially in approximately one-fourth of all subjects with a malignant soft-tissue tumor. Besides the medical problems caused by this situation, the patient loses a chance for cure. When the tumor is a sarcoma, surgery is the basis of treatment. Complementary radiation therapy may be necessary, particularly for high-grade tumors or if the surgical margin was insufficient. Systemic or locoregional chemotherapy can also be used for high-grade or non resectable tumors. PMID- 17124448 TI - [Supracondylar fractures of the humerus in children]. AB - PURPOSE OF THE STUDY: The most frequent type of elbow fracture in children is by far the supracondylar fracture. This type of fracture also raises the greatest risk of nerve injury. We wanted to study the clinical and prognostic features of nerve injuries in children presen-ting supracondylar fractures of the humerus. MATERIAL AND METHODS: We analyzed the files of 55 children with nerve lesions identified among 1 180 files on supracondylar fractures in children. Most presented Lagrange and Ribault stage IV fractures. Nerve injuries involved the radial nerve (n=28), medial nerve (n=20), and ulnar nerve (n=7). The nerve injury was diagnosed before treatment in 32 children, and after treatment in 23. When nerve injury was identified before treatment, closed reduction had been used for eleven fractures and open reduction for 21. Nerve injury identified after treatment was found in eleven fractures after closed reduction and in 12 fractures after open reduction. RESULTS: When nerve injury was recognized at the initial examination, spontaneous nerve recovery was achieved in all cases, irrespective of the treatment modality, within a maximum of four months. When nerve injury was recognized after treatment, spontaneous nerve recovery was obtained in twenty cases. Time to recovery was longer. The three other cases required nerve exploration with neurolysis for two and a nerve graft for one. DISCUSSION: Nerve injury discovered after treatment is either caused by or aggravated by the treatment. Prognosis is less favorable than for injuries discovered at the initial examination. This highlights the importance of carefully searching for nerve deficit, even partial deficiency, in all children presenting a supracondylar fracture of the humerus. It also emphasizes the importance of care in obtaining bone reduction if the initial examination did not reveal any nerve deficit. PMID- 17124449 TI - [Trans-styloid and intrafocal pinning for extra-articular extension fractures of the distal radius: prospective randomized postoperative comparison with simple intra-focal pinning]. AB - PURPOSE OF THE STUDY: Intra-focal pinning as described by Kapandji in 1976 is generally standard treatment of fractures of the distal radius with posterior displacement. This technique, based on the bench effect of pinning on the posterior and radial cortexes has certain limitations in the event of major posterior comminution. In order to improve stability, other authors have proposed a modification of the Kapandji technique, replacing the lateral intra-focal pin with a direct trans-styloid pin. The purpose of this prospective radiographic study was to determine whether the modified Kapandji method improves the quality of postoperative reduction and provides a gain in stability. MATERIAL AND METHODS: This prospective radiographic study included 85 patients. The first group, composed of 41 patients, was treated with the standard intra-focal pinning using one or two dorsal pins and one lateral pin (group K). The second group of 44 patients was treated with one or two intra-focal pins and one lateral transfocal pin (group KM). The postoperative care was the same for the two groups. Clinical and radiographic assessment (lateral and AP views) was performed at day 21, at day 45 after removal of the pins, and at last follow-up. Two operators working independently from the surgeons measured the orientation of the radial surface on the lateral and AP views and the radioulnar variance at day 0, 21, 45 and last follow-up. RESULTS: Group K included 38 patients, mean age 57 years, and group KM 40 patients, mean age 53 years. Seven patients were lost to follow-up. The two groups were comparable regarding age, gender, type of lesion. The postoperative assessment showed radial inclination of 19.3 degrees on the AP view and 3 degrees on the lateral view with 6.6 degrees radiolulnar variance for the K group. The corresponding values were 22.2 degrees , 5.3 degrees and 12.2 degrees for the KM group. Postoperatively, there was a significant difference between the groups for radial glenoid on the AP view (p=0.004) and radiolulnar variance (p<0.0001) but there were no differences on the lateral view. At day 45 postop, mean values were 17.3 degrees AP and 1.1 degrees lateral for radial inclination and 2.2 degrees for radioulnar variance in the K group with 21 degrees , 4.7 degrees and 10 degrees in the KM group. There was a difference in group K for values measured on day 0 and day 45. There was no significant difference between the values on day 0 and day 45 for the KM group. DISCUSSION AND CONCLUSION: The purpose of this radiological assessment was to evaluate the efficacy of replacing the lateral intra-focal pin with a lateral transfocal pin in terms of primary stability of extra-articular fractures of the distal radius with posterior displacement. The results appear to show that this technique provides more sustained reduction during the first six weeks, particularly concerning radioulnar variance and radial glenoid inclination on the AP view, objectives needed for good clinical outcome. It also enables a notable improvement in postoperative stability while preserving the advantages of an easily reproducible minimally destructive percutaneous technique. PMID- 17124450 TI - [Plate fixation with locking screw for distal fractures of the radius]. AB - PURPOSE OF THE STUDY: Fractures of the distal radius are common. No one implant has demonstrated superior efficacy in terms of maintaining the reduction over time. We report our experience with plate fixation using a locking screw. MATERIAL AND METHODS: Between September 2003 and June 2004, 67 displaced fractures of the distal radius were treated by plate fixation using the LCP-DRP 2.4 (Synthes). Three different plates (anterior, posterior, and external) were used. The patients wore a removable anatomic brace for three weeks. Self controlled rehabilitation exercises began directly after surgery with mobilization of the digital chains. The Fernandez, Castaing and AO classifications were noted. Ulnar variance, anteversion of the radial glenoid, radial slope and the alpha angle were measured intraoperatively and at last follow-up to assess maintenance of reduction over time. The DASH test and Green and O'Brien and PRWE scores were used to assess clinical outcome. RESULTS: Mean follow-up was eight months. Mean age was 55.8 years. Eight patients were lost to follow-up. The analysis included 59 patients who could respond to the questionnaires. Bone healing was achieved at six weeks. There were no cases of secondary displacement nor loss of reduction. The Green and O'Brien score was good or very good for 85%. The mean DASH was 20.6 and the mean PRWE 32.8. DISCUSSION: The appropriate fixation method for distal fractures of the radius remains a controversial issue, leading to a variety of materials and fixation methods. Primary stability achieved with the locking screw in a plate enables early mobilization associated with more rapid recovery of function. The absence of secondary displacement, irrespective of the quality of the underlying bone enabled us to achieve equivalent results in young patients and older patients with osteoporotic bone. This study also confirmed the preference for the anterior approach, irrespective of the direction of the displacement. To date, no other material has enabled equivalent results. This is a major achievement in terms of fixation stability. PMID- 17124451 TI - [Axial lower limb alignment and knee geometry in patients with osteoarthritis of the knee]. AB - PURPOSE OF THE STUDY: We conducted a retrospective radiographic study to identify and quantify medial and lateral misalignments in candidates for total knee arthroplasty. MATERIAL AND METHODS: We studied a discontinuous series of 101 patients (164 knees) among candidates for total knee arthroplasty seen between 1990 and 2002. We selected a random sample for study. Exclusion criteria were any history of fracture, surgical treatment, rheumatoid disease, or congenital disorder. A first group of 136 genu varum knees was identified in 82 patients (mean age 72 years, mean weight 79 kg). A second group of 28 genu valgum knees was identified in 19 patients (mean age 67 years, mean weight 71 kg). The standard radiograms were digitalized for semi-automatic measurements using the Metros software. We recorded overall deformation, femoral valgus, tibial varus, HKA angle, HKS angle (mechanical/anatomic axis of the femur) and angle C (horizontal inclination of the ankle to the ground). All measures were compared between each other for each patient to search for relations. RESULTS: In the genu varum group (136 knees), mean overall deformation was 9 degrees , mainly due to tibial misalignment (5.5 degrees ) with a 1 degrees femoral valgus component. The HKS angle was 6.3 degrees , and lateral joint gap 4.3 degrees ; the C angle was 4 degrees . Overall varus followed tibial varus (p<0.01). Ankle inclination followed tibial varus (p<0.01). In the genu valgum group (28 knees), overall deformation was 4.2 degrees , mainly due to femoral misalignment (5.6 degrees) with 1 degrees tibial varus and a medial joint gap of 2 degrees . The HKS angle was 4.7 degrees and the mean C angle -4 degrees . Overall valgus was related to femoral valgus (p<0.001). Tibial varus was greater with greater femoral valgus (p<0.01). In both groups, the HKS angle was smaller with greater femoral valgus. DISCUSSION: Semi-automatic measurements on digitalized films enabled excellent reproducibility. Causes of error were related to limb rotation at acquisition. This study showed that genu varum worsens because of tibial wear followed by ligament distension, while for genu valgum, worsening predominantly results from femoral wear. The ankle joint line was nearly horizontal in one-third of the limbs, even when there was a major misalignment of the lower limb. The HKS angle was quite variable in both morphotypes, but was smaller with greater femoral valgus in both groups. CONCLUSION: Measuring all components involved in medial and lateral knee misalignment is very useful for understanding the morphotype of each individual knee before performing osteotomy or implanting a knee prosthesis. PMID- 17124452 TI - [One-stage reconstruction of the two cruciate ligaments with a single autogenous graft from the extensor system: techniques and results of 25 chronic cases]. AB - PURPOSE OF THE STUDY: Chronic tears of both cruciate ligaments are rarely encountered. Management practices have varied. One-stage reconstruction of both ligaments using arthroscopically implanted allografts was proposed starting in 1996 for such chronic laxity but only 44 cases have been reported in six papers mixed in with recent cases raising very different problems. Treatments using allografts or autogenous grafts or both implanted arthroscopically have often been combined with open surgery. We report a retrospective analysis of 25 knees treated with the same technique from 1983 to 2004: reconstruction with a single transplant taken from the extensor system. MATERIAL AND METHODS: A 30-cm lenght patella tendon-patella-quadriceps tendon graft was harvested. The patellar block was fixed in a hole drilled under the tibial spine, the patellar tendon replacing the posterior cruciate ligament and the quadriceps tendon passing through the lateral condyle and replacing the anterior cruciate ligament before implantation on Gerdy's tubercle, this also creating an extra-articular plasty with the same implant. Medial, lateral and posterior plasties were performed, depending on the radiographic assessment of laxity. Twenty-five knees with major laxity and instability were operated on, including 14 in two-stage operations. The differential anterior drawer was 10+/-3 mm medially and 9+/-5 mm laterally. The posterior drawer at 70 degrees was 15+/-4 mm medially and 10+/-6 degrees laterally. The total anteroposterior laxity of the medial compartment was 29 mm, it was 30 mm for the lateral compartment. Lateral laxity was considered major (>10 degrees ) in 17 knees and medial laxity in 7. Lateral and medial laxities were associated in eleven knees. Hyper-extension was very severe in five knees. RESULTS: Functional outcome at mean 6.5 years follow-up (range 1-21) was satisfactory (except in three knees) allowing renewed occupational activity. Resumption of sports activity was less marked and none of the knees was in IKDC class A, corroborating results in the literature. Anatomic outcome was assessed on the radiological drawer decrease which was modest, mean gain in the anterior drawer measuring 4 and 5 mm in the medial and lateral compartments and mean gain in the posterior drawer measuring 5 mm medially and 3 mm laterally. Peripheral laxity was corrected for best. DISCUSSION: Classically, one or other of the cruciate ligaments is repaired, sometimes both, but in a two-stage procedure associating two different methods and autogenous tendon grafts. One-stage reconstruction of both cruciate ligaments with a single autogenous graft and a wide approach has not led to any complications. One-stage reconstruction using this technique has several advantages: one anterior incision, single transplant which also enables lateral plasty, short immobilization time with rapid rehabilitation. PMID- 17124453 TI - [Results of reimplantation for infected total knee arthroplasty: 107 cases]. AB - PURPOSE OF THE STUDY: The purpose of this study was to assess the results of reimplantations of total knee arthroplasties complicated by infection. Outcome was assessed in terms of eradicated infection and function. MATERIAL AND METHODS: This retrospective multicentric study included 107 cases of infected total knee arthroplasties treated by changing the implants. Seventy-seven patients had a two stage revision and thirty had a one-stage procedure. Patients were reviewed with a minimal 2-year and an average 52-month follow-up. RESULTS: Revision arthroplasty (one- or two-stage) eradicated infection in two out of three patients. With a two-year follow-up, revision arthroplasty was successful in 77% of patients without any sepsis risk factor, in 65% of patients with one risk factor and in 33% of patients with at least two risk factors. After reimplantation for total knee arthroplasty infection, overall function outcome was good (KS knee score: 74.8 after two-stage revision and 75.5 after one-stage revision, NS). After two-stage procedures, the knee outcome was excellent in one third of patients, good in another third and fair or poor in the final third. After one-stage reimplantation, 40% of the knees had an excellent outcome, 30% a good outcome and 30% a fair or poor outcome. Regarding functional outcome, overall results were fair (KS function score 62.5 for one-stage and two-stage revisions). Functional outcome was fair or poor in 42% of patients with a two stage procedure and in 55% of patients with a one-stage revision (NS). DISCUSSION: Our study was unable to disclose any difference between one-stage and two-stage revision for eradicating infection. Unfavorable systemic and local conditions decreased the rate of success after revision total knee arthroplasty for infection. Length of infection before reimplantation, number of surgical procedures and bacterial virulence or resistance were not, in our series, predicting factors for failure of septic revision total knee arthroplasty. No difference was found for the clinical and functional results between one-stage and two-stage procedures. Functional outcome was fair or poor for half of the patients after septic revision total knee arthroplsty. The use of an external device between the two procedures for two-stage revision significantly decreased the functional outcome compared with the use of a spacer. Articulated spacers did not offered any advantage compared with a static spacer for functional outcome. PMID- 17124454 TI - [Long-term behavior of ankle fusion: assessment of the same series at 7 and 23 year (19-36 years) follow-up]. AB - PURPOSE OF THE STUDY: Theoretically, long-term functional and radiographic degradation is predictable after ankle fusion, but sound evidence from the consecutive analysis of the same cohort is lacking. The purpose of this study was to check the hypothesis by repeating assessment in the same cohort of patients who underwent ankle fusion. MATERIAL AND METHODS: The cohort included 52 ankle fusions which had been analyzed in 1984 at seven years (range 2-22 years) follow up then again in 2000 using the same evaluation criteria. Among the 52 patients, six were lost to follow-up (11.5%) 20 had died (38.4%) and one had undergone leg amputation. The second analysis thus included 25 patients (48%). The comparison cohort thus included 25 patients (18 men and 7 women), mean age 62+/-12.6 years (range 40-94) at the 2000 assessment performed 23+/-4.5 years (range 19-36 years) after the fusion. Functional outcome was assessed with the 100-point Duquennoy scale. Osteoarthritis of the subtalar and mediotarsal joints were assessed preoperatively and at follow-up using the same scale. RESULTS: The functional outcome did not deteriorate significantly between 1984 and 2000. The mean score was 65.8+/-22.6 (range 19-92) in 1984 and 64.7+/-18.3 (range 34-90) in 2000 (p=0.67). Fifteen patients (60%) had a good or very good outcome at seven years, and 14 (56%) at 23 years. Between 1984 and 2000, ten patients improved their score (on average 10.4 points, range 1-21 points), two had an unchanged score and thirteen a lower score (on average 10 points, range -1 to -24). Ten of these thirteen patients developed severe intercurrent conditions (neurological or cardiac) explaining the degradation. At last follow-up, sixteen ankles were pain free or nearly pain free. Twelve patients considered their ankle as a forgotten problem (VAS 10) and had no regrets concerning the operation. The evolution of the subtalar joint in 16 cases (nine fusions including five at the same time as the ankle fusion and four performed within four years) showed that all developed osteoarthrtic degradation early with aggravation between 1984 and 2000, leading in the majority of cases to severe degenerative disease. This osteoarthritis was painful in less than one-third of the cases (including the four secondary subtalar fusions and the four subtalar fusions which were painful at mobilization). Twenty-three mediotarsal joints were analyzed (two fusions four years after ankle fusion). The degradation was later and less severe than for the subtalar joint with a majority showing moderate osteoarthritis. Ten ankles exhibited compensatory hypermobility of the forefoot measured at more than 15 degrees without pain. DISCUSSION: This long-term follow-up with two successive assessments using the same evaluation criteria did not demonstrated the late degradation of function expected after ankle fusion. It did show however the presence of undeniable radiographic degradation of the subtalar joint but with little or no severe clinical expression at a minimal follow-up of 19 years. There was no need for complementary fusion between 4 and 23 years follow-up. PMID- 17124455 TI - [Fence grafting for acetabular reconstruction with a cementless cup]. AB - Acetabular reconstruction is difficult after loss of bone stock and socket remodeling. Several techniques have been proposed ranging from a metal backing to allografting. We propose fence grafting. After explantation, the acetabulum is carefully cleaned of all interface tissue and precisely measured. If the vertical diameter is clearly greater than the anteroposterior diameter, a tricortical graft is harvested from the iliac crest and modeled to perfectly fit between the anterioinferior iliac spine and the residual posterior wall as well as the fundus medially. One or two oblique screws are inserted for stabilization. Any superior bone loss is filled by bone substitute (without mechanical value). The acetabulum is then reamed from the obturator foramen sparing the anterior and posterior columns. Residual bony defects are filled with cancellous bone. A hemispheric cup is then press fit and maintained with two or three screws. We performed this procedure in eight patients with SO.F.C.O.T. stage III acetabular loosening with segmentary bone loss and an oval acetabular cavity. Clinical follow-up was more than four years. The Postel-Merle-d'Aubigne score improved from 9.8 to 15.7 on average. Radiographically, there were no implant mobilization or migration and no circumferential lucent lines were observed. A nearly anatomic position was achieved in all cases except two (technical imperfection). At more than one-year follow-up, the grafts could not be distinguished from adjacent bone. For us, high positioned or jumbo cups do not offer a satisfactory reconstruction option. There is a risk of compression with allografts from a head bank. We have not used the cemented metal-backed solution nor impacted grafts. The major drawback with fence grafting is the iliac harvesting (possible residual limping because of the extensive disinsertion of the gluteus medius. The reliable acetabular reconstruction is the major advantage. This technique is not simply an acetabular block widened laterally but it decreases the vertical dimension. This is a reliable but minute technique which allows true long-lasting reconstruction of the acetabulum. PMID- 17124456 TI - [T1-T2 disc herniation: two cases]. AB - We report two cases of exceptional first thoracic disc herniation in a 60-year old man and a 55-year-old woman. The man was treated surgically and the woman medically. Osteoarthritis appeared to be the predominant cause of the disc herniation in both patients. Unlike the usual calcification in the medioposterior position for middle or lower thoracic spine herniations, a soft posterolateral herniation was observed here. The symptoms are limited, as observed in both patients, to a T1 radiculopathy, to be distinguished from C8 radicopathy. Myelopathy is rare. Claude-Bernard-Horner syndrome is not constant but highly suggestive. Both of these signs were absent in our patients. T1-T2 disc herniation should be suspected in patients presenting cervico-brachial medial neuralgia. MRI provides the diagnosis. Anterior surgery can be achieved without sternotomy. Careful radiographic analysis is needed preoperatively to identify the upper limit of the sternum. PMID- 17124457 TI - [Inaugural cutaneous metastases of an osteosarcoma: a case report]. AB - Inaugural cutaneous metastasis is exceptional in osteosarcoma. We report a case of a woman with osteosarcoma of the right knee who presented two tumefactions of the scalp at the time of diagnosis. Magnetic resonance imaging identified a high intensity T2 signal in the femur and also a high-intensity signal in the soft tissues. The head scan confirmed the presence of two subcutaneous tumors. Biopsy of the two sites demonstrated the metastatic nature of the cutaneous lesions. The patient was given preoperative and postoperative chemotherapy. Wide resection followed by prosthesis reconstruction was performed. Pulmonary metastases developed and led to two thoracotomy procedures. The patient has remained in remission 22 months. Six cases of cutaneous metastases of osteosarcoma have been reported in the literature. The cutaneous metastases were secondary in five of six cases, occurring after other metastatic localizations had been discovered. The present case is the first published observation of synchronous cutaneous metastases. The duration of remission suggests that cutaneous localizations should be treated like other inaugural metastatic localizations with intensive chemotherapy and wide resection of the primary tumor. PMID- 17124458 TI - [Chronic expanding hematoma of the soleus: a case report]. AB - Trauma-induced hematomas of the limbs usually resorb without sequelae. In certain circumstances which are not fully understood, the hematoma may expand progressively, eventually leading to the development of a tumor-like mass in the soft tissues. We report the case of a chronic expanding hematoma observed in the right soleus muscle of a 75-year-old man. The mass grew +9 cm compared with the other side over a period of two to three years with no notion of recent trauma. Surgical biopsy disclosed a thick capsule containing "chocolate pus". Pathology and cytology examination led to the diagnosis of pseudo-tumor calcinosis subsequent to a hematoma which the patient had developed 34 years earlier when as a mountain guide he had experienced a tear of the soleus muscle. Local care required complete resection of the soleus muscle. The patient was able to resume activities without pain. Well described in the literature, encapsulated hematoma of the limbs is not well known in France. This case illustrated the potentially long latency period (34 years in our patient). Pathologically similar to tumor calcinosis, chronic expanding hematoma should be entertained as a possible diagnosis in a patient with a longstanding mass and a history of past trauma. The differential diagnosis with sarcoma is established by magnetic resonance imaging which reveals a peripheral low intensity signal on T1 and T2 sequences. PMID- 17124459 TI - [Pneumology, chronic cough and the "Recommendations"]. PMID- 17124460 TI - [Recommendations for clinical practice: chronic cough in the adult]. PMID- 17124465 TI - [Definition, prevalence and measurement methods of chronic cough]. PMID- 17124466 TI - [Current understanding of chronic cough appearing in the ENT context ]. PMID- 17124467 TI - [Current understanding of chronic cough appearing in the broncho-pulmonary context]. PMID- 17124468 TI - [Current understanding of chronic cough appearing in the gastroenterologic context]. PMID- 17124469 TI - [Current understanding of chronic cough appearing in another context: allergic, cardiac, medicamentous, neuropsychiatric, and systemic diseases]. PMID- 17124470 TI - [Diagnostic approach and initial management of cough by general practitioners]. PMID- 17124471 TI - [Diagnostic approach and initial management of cough by the specialist physicians]. PMID- 17124472 TI - [Role and effectiveness of different treatments in chronic cough]. PMID- 17124475 TI - [Imaging of the temporal bone]. PMID- 17124476 TI - [Useful imaging of the ear]. AB - Good anatomical knowledge is necessary to make an effective diagnosis in ear pathology. The purpose of this article is to summarize the main landmarks of the ear. Anatomic definitions of the external ear, middle ear, and bony labyrinth are described on routine spiral CT, and the anatomic definition of the membranous labyrinth and the cochleovestibular nerve on MRI is reviewed. PMID- 17124477 TI - [Imaging in adult chronic otitis]. AB - Chronic otitis media (COM) can be divided into two subtypes: COM with cholesteatoma (including precholesteatomatous states) is an aggressive form of otitis. Surgical treatment is mandatory because of the risk for labyrinthine or cerebromeningeal complications. CT is very important in the preoperative work-up (extension of cholesteatoma, anatomic variants). In patients who have undergone middle ear surgery, CT and presently MRI play an increasing role in the detection of recurrent or relapsing cholesteatoma. COM without cholesteatoma does not have an osteolytic potential, but may leave auditive sequelae that in selected cases may warrant surgical treatment to improve hearing. CT is useful in the etiological work-up of patients with severe hypoacusis. CT also plays an important role in cases of surgical failure, to detect a dislocation of the ossiculoplasty or impairment of the middle ear caused by fluid effusion. The objective of this paper is to specify the indications, the results and the limits of pre- and postoperative imaging in COM. PMID- 17124478 TI - [Otosclerosis imaging: matching clinical and imaging data]. AB - Otosclerosis is a primitive osteodystrophia of the labyrinthine bone. Its diagnosis must be confirmed by a CT scan in order to eliminate the other causes that may lead to conductive hearing loss with an absence of stapedial reflex: fixation of the head of the malleus to the lateral wall of the tympanic cavity, absence of the long process of the incus or stapes, gusher syndrome, primary cholesteatoma, or tympanic facial nerve neuroma blocking the stapes. Particular problems in otosclerosis must be clarified: an extension to the round window (poor postoperative results), and extension to the tympanic cavity blocking the malleus and/or the incus, the labyrinthine lumen, or the internal auditory meatus (very rare). The position of the tympanic facial nerve canal and associated abnormalities must be assessed: stapedial artery, malformations of the ossicles and/or the labyrinth, and chronic otitis media. MRI is indicated in extension to the labyrinthine lumen, the internal auditory meatus, and in postoperative complications with labyrinthitis. MRI can also evaluate the active otosclerotic focus (gadolinium enhancement). PMID- 17124479 TI - [MRI of the temporal bone]. AB - Lesions arising within the temporal bone, where audition and balance receptors are located, are multiple. Beginning with a short clinical and technical overview, this presentation aims to review the most common temporal bone lesions, according to their location. Tumors and malignant otitis externa are the most common lesions of the external auditory canal. MRI features of cholesteatoma redux, transtegmental masses, and paragangliomas are described in the middle ear MRI study. The diagnosis of petrous apex abnormalities is emphasized: cholesterol granuloma, malignant tumor, epidermoid cyst, cholesteatoma, and petrositis. The diagnostic value of CT scan associated with MRI is stressed. This study also includes the main aspects of facial nerve lesions and vascular abnormalities of the area on MRI. The conclusion summarizes the main indications of temporal bone MRI. PMID- 17124480 TI - [The deaf child: anatomy, etiologies and management]. AB - Temporal bone imaging in children shows radioanatomical aspects and diseases distinct from the imaging and pathology results found in adults. Imaging modalities such as CT and MR bring out these differences. The aim of this study is to present the CT and MR particularities of the temporal bone during postnatal growth. The mastoid air cells form mostly in the postnatal period and the course of pneumatization is directly correlated with middle ear successive inflammatory episodes. The most frequent etiologies of hearing loss in children are reviewed, emphasizing their specificities in clinical presentation, radiological aspects, and treatment. In children, conductive hearing loss with normal tympanic membrane is mostly caused by minor aplasia rather than otosclerosis. Sensorineural hearing loss, even when unilateral, is predominantly due to malformation or infection and in rare cases to posterior fossa tumor. PMID- 17124481 TI - The best local therapy for unfavorable risk prostate cancer: the role of surgery. AB - High-risk prostate cancer has an increased rate of local and systemic recurrence after locally definitive therapy. High-risk prostate cancer is defined by using a combination of pretreatment tumor related factors (prostatic specific antigen [PSA] level, stage, Gleason score, and extent of involved biopsy cores) and by pathological findings. Pretreatment PSA kinetics may allow the identification of more patients at high risk of treatment failure who otherwise would have been included in lower risk groups according to conventional risk assignment systems. Eight months of neoadjuvant androgen deprivation therapy prior to radical prostatectomy has been shown in a randomized trial to significantly reduce rates of positive margins compared to 3 months of therapy; however, no significant difference in PSA recurrence rates is apparent 5 years postsurgery. The use of early chemotherapy in prostate cancer has until recently been limited by lack of evidence of an effective chemotherapeutic agent for more advanced disease. Recent data, confirming a survival advantage of docetaxel based regimes in metastatic disease, has focused attention on the use of early chemotherapy in these men with high-risk disease. The technical requirements of surgery on high-risk patients are now better defined and one challenge for the specialty is to take this knowledge and apply it successfully in the laparoscopic setting. However, the limit of surgery alone in reducing recurrence in high-risk disease from technical advancements has plateaued. In order to take the field forward, successful multimodal treatment strategies are needed to improve the outcomes over surgical monotherapy for high-risk disease. Novel nucleotide therapy targeting the production of cell survival proteins has provided promising phase 1 data in prostate cancer. This experimental therapy has been built on an understanding of the observed effects that androgen deprivation therapy has on cancer cell survival proteins produced during periods of cellular stress. PMID- 17124482 TI - Androgen deprivation therapy in locally advanced and metastatic prostate cancer. AB - Numerous studies have been performed testing the efficacy of early androgen deprivation (AD) in patients with localized and locally advanced prostate cancer. A systematic review of recent publications that report on the use of AD in non metastatic prostate cancer patients was performed. Recently published mature randomized trials of AD plus local therapy were evaluated plus 2 large datasets on the use of AD for patients with serologic relapse after local therapy. Four mature randomized studies demonstrate an overall survival benefit to the use of AD in conjunction with definitive local therapy (3 with radiation and 1 with surgery). One retrospective analysis suggests that AD administered at early after serologic progression improves overall survival, and one retrospective analysis shows a reduction in metastasis-free survival but has not yet shown an overall survival benefit. In virtually all analyses, patients with high-risk features benefited from early AD when compared to deferred therapy. Consideration of AD is therefore warranted early in the clinical course of high-risk patients. PMID- 17124483 TI - Renovascular disease: a review of diagnostic and therapeutic procedures. AB - The clinical importance of renovascular disease, atherosclerotic or of other origin, arises from the fact, that renal artery stenosis (RAS), if hemodynamically significant (> 70% diameter reduction), induces arterial hypertension, renal insufficiency or both. The prevalence of RAS rises with increasing age and with the presence of atherosclerosis of the aorta, carotid, coronary and peripheral arteries. Typical clinical symptoms, as uncontrolled hypertension or renal dysfunction in the absence of pathological urinary findings, are helpful to select patients for further screening methods: We see a prominent role of color duplex sonography as a screening procedure. Intra arterial angiography remains gold standard for the diagnosis of RAS. The major problem in daily clinical practice is the differentiation between patients in which hypertension and kidney function can be improved or normalized by removal of RAS and those with ''fixed'' hypertension and irreversible kidney dysfunction and therefore to decide if it is worth while to perform invasive treatment as angioplasty or surgery. In this setting, the proof of hemodynamic significance is essential and is indicated especially when the stenosis has a diameter reduction of < 50-70% only. Methods proving a critical stenosis are intra-arterial measurement of the pressure gradient, measurement of differential renal vein renin and duplex sonography. In addition, predictors of treatment outcome should be considered. Studies analyzing if patients improve with blood pressure and kidney function after removal of RAS have shown that high grade stenosis and/or very high blood pressure indicate a good outcome. Further prognostic factors are the absence of parenchymal disease and/or positive functional test. In the presence of a critical stenosis in a patient with a clear clinical problem with hypertension and/or renal dysfunction a positive effect of invasive treatment seems warranted despite the risks that must be considered as well in angioplasty as in surgery. The selection for the type of invasive treatment requires a clarification of the treatment goals in the individual patient, the evaluation of the morphology and localization of the stenosis as the presence of other vascular disease (aortic aneurysm, peripheral artery disease etc.) and the assessment of the risk according to the type of intervention. PMID- 17124484 TI - Testicular hypotrophy in varicocele: pre and postoperative echographic follow-up in the pediatric age. AB - AIM: Varicocele is characterised by an anomalous tortuosity and dilation of the veins of the pampiniform plexus. The etiopathogenetic mechanisms are still unclear, but a correlation seems to exist between varicocele and testicular development, with possible repercussions on the testicle's functionality. The aim of this study is to evaluate gonadic trophism through echographic monitoring in the pre and postoperative phases in patients affected by idiopathic varicocele with testicular hypotrophy in order to evaluate the treatment's benefits. METHODS: Sixty-six patients (mean age 12,5; range 10-17) consecutively operated for left idiopathic varicocele were considered. Of these, 27 had ipsilateral testicular hypotrophy and thus they were included in the study. Fifteen were operated upon in videolaparoscopy (VLS), and 12 by the classic open inguinal access. The mean follow-up was 18 months (6-24 months). The data were analyzed by nonparametric Mann-Whitney U test. RESULTS: An increase in the testicular volume was observed clinically and by ultrasound in 13 of the 15 patients treated by VLS and in 9 of the 12 patients operated by traditional means. The nonparametric Mann Whitney U test showed a significativity between pre and post-operative values. CONCLUSION: The testicular trophic healing observed in 81.5% of the operated patients leads to the belief that an early correction can allow a rapid volumetric increase and an improved function of the gonad. PMID- 17124485 TI - Diagnostic imaging of inverted Y duplication of the ureter with anomalous point angle. A case report. AB - A rare anomaly of ureteral duplication is the inverted Y configuration, occurring when 2 distal ureteral limbs fuse proximally to become a single tube draining the kidney. In international literature there is a female predominance. Previous reports documented distal limbs that were atresic or associated with ureterocele or ectopically located. We report a case of inverted Y ureter with an obtuse point angle open caudally; the 2 ureteral limbs reach the bladder trigone and are canalized, even if the supernumerary one is partially stenotic in the distal tract. PMID- 17124486 TI - Extragastrointestinal stromal tumor of the retroperitoneum mimicking kidney tumor. AB - A case of extragastrointestinal tumor of the retroperitoneum in a 48-year-old woman complaining right sided flank pain without hematuria is reported. The mass was excised from the kidney without a positive margin. The histopathological examination revealed an extragastrointestinal stromal tumor of the retroperitoneum. These tumors usually originate from the small intestine or stomach. On the other hand, they are rarely located in the retroperitoneum. These tumors typically exhibit CD117 immunoreactivity, whereas they may reveal CD34, neuron specific enolase, smooth muscle actin, desmin and S-100 protein. In our case the specimen of the patient was positive for CD117, actin and desmin. PMID- 17124487 TI - Nephro-ileal conduit subcutaneous stent: an alternative to percutaneous nephrostomy. AB - Nephrostomy has been the standard method of urinary diversion for chronic ureteral obstruction or when placement of ureteral stents has failed. We describe the first case of left kidney ileal conduit (Briker's urinary diversion) subcutaneous urinary by-pass. This method made the patient free from a second collecting bag without any significant complications. This method may improve the quality of life of patients with severe ureteral obstruction and ileal conduit. PMID- 17124488 TI - One step endourologic management of severely encrusted stent in solitary kidney with renal impairment. A case report. AB - Removing a severely encrusted stent, without an initial preventive descaling of the calcification that envelopes it, can lead to serious complications such as the breaking up of the stent, lesions and ureteral avulsion. We report a case regarding an entirely endourological treatment of an encrusted stent in a single kidney patient with renal insufficiency. The time the stent had been in place was 6 months. The patient underwent bladder lithotripsy of the distal tip of the encrusted stent, ureterolithotripsy to free the intraureteral tract of any incrustation and percutaneous nephrolithotomy to deal with the stone burden around the proximal tip. The stent, once free from fragments, was removed, via anterograde, without causing any friction in the excretory tract. The anesthesia time was 3.5 h. The postoperative period was normal without any septic complications or bleeding. We observed a slight worsening of the creatininemia which, after 3 days, could be compared with the preoperative blood levels. Ten days after the operation, renal function returned to normal. Even for patients with just one kidney and renal insufficiency the endourological management of encrusted stents does not lead to a high morbidity and is quite feasible as a one step operation. PMID- 17124489 TI - [Adenocarcinoma of the scrotum: a case report]. PMID- 17124490 TI - [Leiomyosarcoma of the vessels of the renal ileus : a case report]. PMID- 17124491 TI - Prepubertal testis development relies on retinoic acid but not rexinoid receptors in Sertoli cells. AB - Sertoli cells (SC) are instrumental to stem spermatogonia differentiation, a process that critically depends on retinoic acid (RA). We show here that selective ablation of RA receptor alpha (RARalpha) gene in mouse SC, singly (Rara(Ser-/-) mutation) or in combination with RARbeta and RARgamma genes (Rara/b/g(Ser-/-) mutation), abolishes cyclical gene expression in these cells. It additionally induces testis degeneration and delays spermatogonial expression of Stra8, two hallmarks of RA deficiency. As identical defects are generated upon inactivation of RARalpha in the whole organism, our data demonstrate that all the functions exerted by RARalpha in male reproduction are Sertoli cell-autonomous. They further indicate that RARalpha is a master regulator of the cyclical activity of SC and controls paracrine pathways required for spermatogonia differentiation and germ cell survival. Most importantly, we show that the ablation of all RXR (alpha, beta and gamma isotypes) in SC does not recapitulate the phenotype generated upon ablation of all three RARs, thereby providing the first evidence that RARs exert functions in vivo independently of RXRs. PMID- 17124492 TI - ATR-dependent phosphorylation and activation of ATM in response to UV treatment or replication fork stalling. AB - The phosphatidyl inositol 3-kinase-like kinases (PIKKs), ataxia-telangiectasia mutated (ATM) and ATM- and Rad3-related (ATR) regulate parallel damage response signalling pathways. ATM is reported to be activated by DNA double-strand breaks (DSBs), whereas ATR is recruited to single-stranded regions of DNA. Although the two pathways were considered to function independently, recent studies have demonstrated that ATM functions upstream of ATR following exposure to ionising radiation (IR) in S/G2. Here, we show that ATM phosphorylation at Ser1981, a characterised autophosphorylation site, is ATR-dependent and ATM-independent following replication fork stalling or UV treatment. In contrast to IR-induced ATM-S1981 phosphorylation, UV-induced ATM-S1981 phosphorylation does not require the Nbs1 C-terminus or Mre11. ATR-dependent phosphorylation of ATM activates ATM phosphorylation of Chk2, which has an overlapping function with Chk1 in regulating G2/M checkpoint arrest. Our findings provide insight into the interplay between the PIKK damage response pathways. PMID- 17124493 TI - Huntingtin inhibits caspase-3 activation. AB - Huntington's disease results from a mutation in the HD gene encoding for the protein huntingtin. The function of huntingtin, although beginning to be elucidated, remains largely unclear. To probe the prosurvival function of huntingtin, we modulate levels of wild-type huntingtin in a number of cellular and in vivo models. Huntingtin depletion resulted in caspase-3 activation, and overexpression of huntingtin resulted in caspase-3 inhibition. Additionally, we demonstrate that huntingtin physically interacts with active caspase-3. Interestingly, mutant huntingtin binds active caspase-3 with a lower affinity and lower inhibitory effect on active caspase-3 than does wild-type huntingtin. Although reduction of huntingtin levels resulted in caspase-3 activation in all conditions examined, the cellular response was cell-type specific. Depletion of huntingtin resulted in either overt cell death, or in increased vulnerability to cell death. These data demonstrate that huntingtin inhibits caspase-3 activity, suggesting a mechanism whereby caspase-mediated huntingtin depletion results in a detrimental amplification cascade leading to further caspase-3 activation, resulting in cell dysfunction and cell death. PMID- 17124494 TI - A synchronized substrate-gating mechanism revealed by cubic-core structure of the bovine branched-chain alpha-ketoacid dehydrogenase complex. AB - The dihydrolipoamide acyltransferase (E2b) component of the branched-chain alpha ketoacid dehydrogenase complex forms a cubic scaffold that catalyzes acyltransfer from S-acyldihydrolipoamide to CoA to produce acyl-CoA. We have determined the first crystal structures of a mammalian (bovine) E2b core domain with and without a bound CoA or acyl-CoA. These structures reveal both hydrophobic and the previously unreported ionic interactions between two-fold-related trimers that build up the cubic core. The entrance of the dihydrolipoamide-binding site in a 30-A long active-site channel is closed in the apo and acyl-CoA-bound structures. CoA binding to one entrance of the channel promotes a conformational change in the channel, resulting in the opening of the opposite dihydrolipoamide gate. Binding experiments show that the affinity of the E2b core for dihydrolipoamide is markedly increased in the presence of CoA. The result buttresses the model that CoA binding is responsible for the opening of the dihydrolipoamide gate. We suggest that this gating mechanism synchronizes the binding of the two substrates to the active-site channel, which serves as a feed-forward switch to coordinate the E2b-catalyzed acyltransfer reaction. PMID- 17124495 TI - Identification of a strong binding site for kinesin on the microtubule using mutant analysis of tubulin. AB - The kinesin-binding site on the microtubule has not been identified because of the technical difficulties involved in the mutant analyses of tubulin. Exploiting the budding yeast expression system, we succeeded in replacing the negatively charged residues in the alpha-helix 12 of beta-tubulin with alanine and analyzed their effect on kinesin-microtubule interaction in vitro. The microtubule gliding assay showed that the affinity of the microtubules for kinesin was significantly reduced in E410A, D417A, and E421A, but not in E412A mutant. The unbinding force measurement revealed that in the former three mutants, the kinesin-microtubule interaction in the adenosine 5'-[beta,gamma-imido]triphosphate state (AMP-PNP state) became less stable when a load was imposed towards the microtubule minus end. In parallel with this decreased stability, the stall force of kinesin was reduced. Our results implicate residues E410, D417, and E421 as crucial for the kinesin-microtubule interaction in the strong binding state, thereby governing the size of kinesin stall force. PMID- 17124496 TI - TWEAK, via its receptor Fn14, is a novel regulator of mesenchymal progenitor cells and skeletal muscle regeneration. AB - Inflammation participates in tissue repair through multiple mechanisms including directly regulating the cell fate of resident progenitor cells critical for successful regeneration. Upon surveying target cell types of the TNF ligand TWEAK, we observed that TWEAK binds to all progenitor cells of the mesenchymal lineage and induces NF-kappaB activation and the expression of pro-survival, pro proliferative and homing receptor genes in the mesenchymal stem cells, suggesting that this pro-inflammatory cytokine may play an important role in controlling progenitor cell biology. We explored this potential using both the established C2C12 cell line and primary mouse muscle myoblasts, and demonstrated that TWEAK promoted their proliferation and inhibited their terminal differentiation. By generating mice deficient in the TWEAK receptor Fn14, we further showed that Fn14 deficient primary myoblasts displayed significantly reduced proliferative capacity and altered myotube formation. Following cardiotoxin injection, a known trigger for satellite cell-driven skeletal muscle regeneration, Fn14-deficient mice exhibited reduced inflammatory response and delayed muscle fiber regeneration compared with wild-type mice. These results indicate that the TWEAK/Fn14 pathway is a novel regulator of skeletal muscle precursor cells and illustrate an important mechanism by which inflammatory cytokines influence tissue regeneration and repair. Coupled with our recent demonstration that TWEAK potentiates liver progenitor cell proliferation, the expression of Fn14 on all mesenchymal lineage progenitor cells supports a broad involvement of this pathway in other tissue injury and disease settings. PMID- 17124497 TI - Dual binding sites for translocation catalysis by Escherichia coli glutathionylspermidine synthetase. AB - Most organisms use glutathione to regulate intracellular thiol redox balance and protect against oxidative stress; protozoa, however, utilize trypanothione for this purpose. Trypanothione biosynthesis requires ATP-dependent conjugation of glutathione (GSH) to the two terminal amino groups of spermidine by glutathionylspermidine synthetase (GspS) and trypanothione synthetase (TryS), which are considered as drug targets. GspS catalyzes the penultimate step of the biosynthesis-amide bond formation between spermidine and the glycine carboxylate of GSH. We report herein five crystal structures of Escherichia coli GspS in complex with substrate, product or inhibitor. The C-terminal of GspS belongs to the ATP-grasp superfamily with a similar fold to the human glutathione synthetase. GSH is likely phosphorylated at one of two GSH-binding sites to form an acylphosphate intermediate that then translocates to the other site for subsequent nucleophilic addition of spermidine. We also identify essential amino acids involved in the catalysis. Our results constitute the first structural information on the biochemical features of parasite homologs (including TryS) that underlie their broad specificity for polyamines. PMID- 17124498 TI - A Cdt1-geminin complex licenses chromatin for DNA replication and prevents rereplication during S phase in Xenopus. AB - Initiation of DNA synthesis involves the loading of the MCM2-7 helicase onto chromatin by Cdt1 (origin licensing). Geminin is thought to prevent relicensing by binding and inhibiting Cdt1. Here we show, using Xenopus egg extracts, that geminin binding to Cdt1 is not sufficient to block its activity and that a Cdt1 geminin complex licenses chromatin, but prevents rereplication, working as a molecular switch at replication origins. We demonstrate that geminin is recruited to chromatin already during licensing, while bulk geminin is recruited at the onset of S phase. A recombinant Cdt1-geminin complex binds chromatin, interacts with the MCM2-7 complex and licenses chromatin once per cell cycle. Accordingly, while recombinant Cdt1 induces rereplication in G1 or G2 and activates an ATM/ATR dependent checkpoint, the Cdt1-geminin complex does not. We further demonstrate that the stoichiometry of the Cdt1-geminin complex regulates its activity. Our results suggest a model in which the MCM2-7 helicase is loaded onto chromatin by a Cdt1-geminin complex, which is inactivated upon origin firing by binding additional geminin. This origin inactivation reaction does not occur if only free Cdt1 is present on chromatin. PMID- 17124499 TI - Protein kinase B/Akt phosphorylation of PDE3A and its role in mammalian oocyte maturation. AB - cGMP-inhibited cAMP phosphodiesterase 3A (PDE3A) is expressed in mouse oocytes, and its function is indispensable for meiotic maturation as demonstrated by genetic ablation. Moreover, PDE3 activity is required for insulin/insulin-like growth factor-1 stimulation of Xenopus oocyte meiotic resumption. Here, we investigated the cAMP-dependent protein kinase B (PKB)/Akt regulation of PDE3A and its impact on oocyte maturation. Cell-free incubation of recombinant mouse PDE3A with PKB/Akt or cAMP-dependent protein kinase A catalytic subunits leads to phosphorylation of the PDE3A protein. Coexpression of PDE3A with constitutively activated PKB/Akt (Myr-Akt) increases PDE activity as well as its phosphorylation state. Injection of pde3a mRNA potentiates insulin-dependent maturation of Xenopus oocytes and rescues the phenotype of pde3(-/-) mouse oocytes. This effect is greatly decreased by mutation of any of the PDE3A serines 290-292 to alanine in both Xenopus and mouse. Microinjection of myr-Akt in mouse oocytes causes in vitro meiotic maturation and this effect requires PDE3A. Collectively, these data indicate that activation of PDE3A by PKB/Akt-mediated phosphorylation plays a role in the control of PDE3A activity in mammalian oocytes. PMID- 17124500 TI - Sphingosine 1-phosphate as a regulator of osteoclast differentiation and osteoclast-osteoblast coupling. AB - Sphingosine 1-phosphate (S1P), produced by sphingosine kinase (SPHK), acts both by intracellular and extracellular modes. We evaluated the role of SPHK1 and S1P in osteoclastogenesis using bone marrow-derived macrophage (BMM) single and BMM/osteoblast coculture systems. In BMM single cultures, the osteoclastogenic factor receptor activator of NF-kappaB ligand (RANKL) upregulated SPHK1 and increased S1P production and secretion. SPHK1 siRNA enhanced and SPHK1 overexpression attenuated osteoclastogenesis via modulation of p38 and ERK activities, and NFATc1 and c-Fos levels. Extracellular S1P had no effect in these cultures. These data suggest that intracellular S1P produced in response to RANKL forms a negative feedback loop in BMM single cultures. In contrast, S1P addition to BMM/osteoblast cocultures greatly increased osteoclastogenesis by increasing RANKL in osteoblasts via cyclooxygenase-2 and PGE(2) regulation. S1P also stimulated osteoblast migration and survival. The RANKL elevation and chemotactic effects were also observed with T cells. These results indicate that secreted S1P attracts and acts on osteoblasts and T cells to augment osteoclastogenesis. Taken together, S1P plays an important role in osteoclastogenesis regulation and in communication between osteoclasts and osteoblasts or T cells. PMID- 17124501 TI - Redundant functions of RIM1alpha and RIM2alpha in Ca(2+)-triggered neurotransmitter release. AB - Alpha-RIMs (RIM1alpha and RIM2alpha) are multidomain active zone proteins of presynaptic terminals. Alpha-RIMs bind to Rab3 on synaptic vesicles and to Munc13 on the active zone via their N-terminal region, and interact with other synaptic proteins via their central and C-terminal regions. Although RIM1alpha has been well characterized, nothing is known about the function of RIM2alpha. We now show that RIM1alpha and RIM2alpha are expressed in overlapping but distinct patterns throughout the brain. To examine and compare their functions, we generated knockout mice lacking RIM2alpha, and crossed them with previously produced RIM1alpha knockout mice. We found that deletion of either RIM1alpha or RIM2alpha is not lethal, but ablation of both alpha-RIMs causes postnatal death. This lethality is not due to a loss of synapse structure or a developmental change, but to a defect in neurotransmitter release. Synapses without alpha-RIMs still contain active zones and release neurotransmitters, but are unable to mediate normal Ca(2+)-triggered release. Our data thus demonstrate that alpha-RIMs are not essential for synapse formation or synaptic exocytosis, but are required for normal Ca(2+)-triggering of exocytosis. PMID- 17124502 TI - CaMKII translocation requires local NMDA receptor-mediated Ca2+ signaling. AB - Excitatory synaptic transmission and plasticity are critically modulated by N methyl-D-aspartate receptors (NMDARs). Activation of NMDARs elevates intracellular Ca(2+) affecting several downstream signaling pathways that involve Ca(2+)/calmodulin-dependent protein kinase II (CaMKII). Importantly, NMDAR activation triggers CaMKII translocation to synaptic sites. NMDAR activation failed to induce Ca(2+) responses in hippocampal neurons lacking the mandatory NMDAR subunit NR1, and no EGFP-CaMKIIalpha translocation was observed. In cells solely expressing Ca(2+)-impermeable NMDARs containing NR1(N598R)-mutant subunits, prolonged NMDA application elevated internal Ca(2+) to the same degree as in wild-type controls, yet failed to translocate CaMKIIalpha. Brief local NMDA application evoked smaller Ca(2+) transients in dendritic spines of mutant compared to wild-type cells. CaMKIIalpha mutants that increase binding to synaptic sites, namely CaMKII-T286D and CaMKII-TT305/306VA, rescued the translocation in NR1(N598R) cells in a glutamate receptor-subtype-specific manner. We conclude that CaMKII translocation requires Ca(2+) entry directly through NMDARs, rather than other Ca(2+) sources activated by NMDARs. Together with the requirement for activated, possibly ligand-bound, NMDARs as CaMKII binding partners, this suggests that synaptic CaMKII accumulation is an input specific signaling event. PMID- 17124503 TI - Piebald trait: implication of kit mutation on in vitro melanocyte survival and on the clinical application of cultured epidermal autografts. AB - Piebald trait leukoderma results from "loss-of-function" mutations in the kit gene. Correlations between mutation type and clinical phenotype have been reported. However, mutation classification has been mainly based on the clinical features of patients. The aim of this study was to get a better understanding of the pathogenesis of human piebaldism by establishing whether the kit mutation type may affect the in vitro survival/proliferation of patient melanocytes. Overall, the research was finalized to implement the clinical application of the autologous cultured epidermis in the treatment of piebald patients. Seven patients, who were transplanted with autologous in vitro reconstituted epidermis, showed an average percentage of repigmentation of 90.7. Six novel and one previously reported mutations were found and their postulated effects discussed in relation to the clinical phenotype and in vitro behavior of epidermal cells. Although mutation type did not impair repigmentation given by autotransplantation, it was shown to influence the survival/proliferation of co cultured melanocytes and keratinocytes. In particular, tyrosine kinase domain mutations were found with melanocyte loss and keratinocyte senescence during expansion of epidermal cultures. Results indicate that the clinical application of cultured epidermis in piebald patients may be optimized by investigating mutation functional effects before planning surgical operations. PMID- 17124504 TI - A skin-like cytochrome P450 cocktail activates prohaptens to contact allergenic metabolites. AB - Allergic contact dermatitis is a complex syndrome representing immunological responses to cutaneous exposure to protein-reactive chemicals. Although many contact sensitizers directly can elicit this disorder, others (prohaptens) require activation. Knowledge regarding the activating mechanisms remains limited, but one possibility is metabolic activation by cytochrome P450 (CYP) enzymes in the skin. We have, after quantitative reverse transcriptase-PCR studies of the CYP content in 18 human skin samples, developed an enriched skin like recombinant human (rh) CYP cocktail using CYP1A1, 1B1, 2B6, 2E1, and 3A5. To validate the rhCYP cocktail, a prohaptenic conjugated diene ((5R)-5-isopropenyl-2 methyl-1-methylene-2-cyclohexene) was investigated using: the skin-like rhCYP cocktail, a liver-like rhCYP cocktail, single rhCYP enzymes, liver microsomes, keratinocytes, and a dendritic cell (DC) assay. The diene was activated to sensitizing epoxides in all non-cell-based incubations including the skin-like rhCYP cocktail. An exocyclic epoxide metabolite ((7R)-7-isopropenyl-4-methyl-1 oxaspiro[2.5]oct-4-ene) was found to be mainly responsible for the allergenic activity of the diene. This epoxide also induced pronounced DC activation indicated by upregulation of IL-8. The skin-like rhCYP cocktail provides a simplified alternative to using skin tissue preparations in mechanistic studies of CYP-mediated skin metabolism of prohaptens and offers the future possibility of designing in vitro predictive assays for assessment of allergenic activity of prohaptens. PMID- 17124505 TI - Modulation of porcine wound repair with a transfected ErbB3 gene and relevant EGF like ligands. AB - Our in vivo study used an ErbB3 receptor transfection strategy to determine if topical application of EGF-like ligands would enhance repair. Partial-thickness porcine wounds transfected with adenoviral particles containing an ErbB3 receptor gene or a vehicle beta-galactosidase gene were introduced and wounds were concomitantly supplied with a variety of EGF-like ligands--EGF, epiregulin (EPR), heparin binding EGF (HB-EGF), and heregulin/neuregulin (HRG). Comparisons of cutaneous repair (resurfacing, dermal depth, proliferation, macrophage infiltration, microvascular density, apoptosis) were assessed after a 5-day healing interval. Differential effects were noted. In wounds transfected with additional ErbB3, either EPR or HB-EGF promoted resurfacing greater than EGF, HRG, or controls. Dermal responses differed significantly after EPR or HB-EGF treatments compared to EGF, HRG, ErbB3 only, or empty vehicle. Hallmarks of enhanced wound maturity were noted in EPR- and HB-EGF-treated wounds transfected with ErbB3. Our data confirmed that an ErbB3-driven pathway mediates a net positive influence in an in vivo model closely resembling human repair. The sensitivity in this system was sufficient to reveal differential outcomes following stimulation with various EGF ligands. We conclude that selective stimulation through an ErbB3-driven pathway shows promise as a therapeutic strategy to hasten wound maturity. PMID- 17124506 TI - The role of the calcium sensing receptor in regulating intracellular calcium handling in human epidermal keratinocytes. AB - Calcium is critical for controlling the balance of proliferation and differentiation in epidermal keratinocytes. We previously reported that the calcium sensing receptor (CaR) is required for mediating Ca2+ signaling and extracellular Ca2+ (Ca2+(o))-induced differentiation. In this study, we investigated the mechanism by which CaR regulates intracellular Ca2+ (Ca2+(i)) and its role in differentiation. Membrane fractionation, fluorescence immunolocalization, and co-immunoprecipitation studies were performed to assess potential interactions between CaR and other regulators of Ca2+ stores and channels. We found that the glycosylated form of CaR forms a complex with phospholipase C gamma1, IP3 receptor (IP3R), and the Golgi Ca2+-ATPase, secretory pathway Ca2+-ATPase 1, in the trans-Golgi. Inactivation of the endogenous CaR gene by adenoviral expression of a CaR antisense cDNA inhibited Ca2+(i) response to Ca2+(o), decreased Ca2+(i) stores, decreased Ca2+(o)-induced differentiation, but augmented store-operated channel activity and Ca2+ uptake by intracellular organelles. Our results indicate that CaR regulates keratinocyte differentiation in part by modulating Ca2+(i) stores via interactions with Ca2+ pumps and channels that regulate those stores. PMID- 17124507 TI - Novel mutations in the BHD gene and absence of loss of heterozygosity in fibrofolliculomas of Birt-Hogg-Dube patients. AB - Birt-Hogg-Dube (BHD) syndrome is an autosomal-dominantly inherited cancer syndrome characterized by fibrofolliculomas, lung cysts leading to pneumothorax, and chromophobic/oncocytic renal cell carcinoma. The disease is caused by heterozygous mutations in the BHD gene encoding folliculin and all mutations reported putatively lead to protein truncation. Although the function of folliculin is unknown, it is thought to be a tumor suppressor, with loss of heterozygosity (LOH) initiating tumor formation. Here, we report on four novel BHD gene mutations, including two splice-site mutations, in patients presenting with skin lesions only. We further show that LOH cannot be detected in fibrofolliculomas from three patients, suggesting that for the manifestation of cutaneous tumors in BHD syndrome haplo-insufficiency of folliculin is sufficient to initiate uncontrolled growth. Renal microscopic oncocytosis in BHD is considered as a precursor to malignant kidney tumors and may likewise be the result of haplo-insufficiency, with somatic second-hit mutations or LOH giving rise to malignancy later in life. PMID- 17124508 TI - A novel MASH1 enhancer with N-myc and CREB-binding sites is active in neuroblastoma. AB - Neuroblastoma is one of the most common solid tumors in childhood. With the aim of developing a targeting vector for neuroblastoma, we cloned and characterized an enhancer in the 5'-flanking regions of the MASH1 gene by a random-trap method from a 36 kb cosmid DNA. The enhancer-containing clone was identified by the expression of GFP when transfected into neuroblastoma cell lines. The enhancer luciferase activity is higher in neuroblastoma cell lines, IMR32, BE2 and SH SY5Y, compared with those in non-neuroblastoma cell lines, U1242 glioma, N417 small cell lung cancer and EOMA hemangioma. The core enhancer was determined within a 0.2 kb fragment, yielding three- to fourfold higher activity than that of the MASH1 promoter alone in IMR32 and BE2. This area possesses GATA- and CREB binding sites, as well as the E-box. EMSA on this area demonstrated that CREB/ATF could bind the DNA. Chromatin immunoprecipitation assay revealed that N-myc, CREB, and co-activators CBP and PCAF, but not HDAC1, are bound to the core enhancer at the same time as the co-activators and N-myc bind to the promoter. This supports the idea that the commonly overexpressed genes HASH1 and N-myc are regulated in concert, confirming their importance as prognostic markers or targets for therapy. PMID- 17124509 TI - Humoral and cellular immunity induced by tumor cell vaccine based on the chicken xenogeneic homologous matrix metalloproteinase-2. AB - Matrix metalloproteinase-2 (MMP-2) has been used as a target for cancer immunotherapy. The activation of immunization by breaking immune tolerance to self-MMP-2 may be one of the promising approaches for the treatment of MMP-2 positive tumors. In this study, we constructed the xenogeneic tumor cell vaccine c-MMP-2 by transfecting CT26 and LLC cells with chicken MMP-2 cDNA constructs. MMP-2-specific autoantibodies in sera and tumor cells were found in mice immunized with c-MMP-2. Protection against tumor growth was evaluated in respect of the relative contributions of autoantibodies, CD4+, and CD8+ T cells. Treatment with this vaccine (c-MMP-2) also prolonged the survival time of mice bearing cancer. The specific cytotoxic T-cell responses suggested that the treatment increased CD8+ T-cell activity. The antitumor activity of c-MMP-2 was abrogated by in vivo depletion of CD4+ and CD8+ T-lymphocytes and improved by adoptive transfer of CD4+ and CD8+ T-lymphocytes from the mice treated with c-MMP 2. An alternative DNA vaccination strategy for cancer therapy was identified in this study by eliciting humoral and cellular immunoresponse with a crossreacting transfectant. PMID- 17124510 TI - Snail genes at the crossroads of symmetric and asymmetric processes in the developing mesoderm. AB - Retinoic acid (RA) signalling ensures that vertebrate mesoderm segmentation is bilaterally synchronized, and corrects transient interferences from asymmetric left-right (L-R) signals involved in organ lateralization. Snail genes participate in both these processes and, although they are expressed symmetrically in the presomitic mesoderm (PSM), Snail1 transcripts are asymmetrically distributed in the L-R lateral mesoderm. We show that the alteration of the symmetric Snail expression in the PSM induces asynchronous somite formation. Furthermore, in the absence of RA signalling, normal asymmetric Snail1 expression in the lateral mesoderm is extended to the PSM, desynchronizing somitogenesis. Thus, Snail1 is the first cue corrected by RA in the PSM to ensure synchronized bilateral segmentation. PMID- 17124511 TI - The supramolecular architecture of junctional microdomains in native lens membranes. AB - Gap junctions formed by connexons and thin junctions formed by lens-specific aquaporin 0 (AQP0) mediate the tight packing of fibre cells necessary for lens transparency. Gap junctions conduct water, ions and metabolites between cells, whereas junctional AQP0 seems to be involved in cell adhesion. High-resolution atomic force microscopy (AFM) showed the supramolecular organization of these proteins in native lens core membranes, in which AQP0 forms two-dimensional arrays that are surrounded by densely packed gap junction channels. These junctional microdomains simultaneously provide adhesion and communication between fibre cells. The AFM topographs also showed that the extracellular loops of AQP0 in junctional microdomains adopt a conformation that closely resembles the structure of junctional AQP0, in which the water pore is thought to be closed. Finally, time-lapse AFM imaging provided insights into AQP0 array formation. This first high-resolution view of a multicomponent eukaryotic membrane shows how membrane proteins self-assemble into functional microdomains. PMID- 17124512 TI - Tuning inflammation and immunity by chemokine sequestration: decoys and more. AB - A set of chemokine receptors are structurally unable to elicit migration or conventional signalling responses after ligand engagement. These 'silent' (non signalling) chemokine receptors regulate inflammatory and immune reactions in different ways, including by acting as decoys and scavengers. Chemokine decoy receptors recognize distinct and complementary sets of ligands and are strategically expressed in different cellular contexts. Importantly, viruses and parasites have evolved multiple strategies to elude chemokines, including the expression of decoy receptors. So, decoy receptors for chemokines represent a general strategy to tune, shape and temper innate and adaptive immunity. PMID- 17124513 TI - SNAREing immunity: the role of SNAREs in the immune system. AB - The trafficking of molecules and membranes within cells is a prerequisite for all aspects of cellular immune functions, including the delivery and recycling of cell-surface proteins, secretion of immune mediators, ingestion of pathogens and activation of lymphocytes. SNARE (soluble-N-ethylmaleimide-sensitive-factor accessory-protein receptor)-family members mediate membrane fusion during all steps of trafficking, and function in almost all aspects of innate and adaptive immune responses. Here, we provide an overview of the roles of SNAREs in immune cells, offering insight into one level at which precision and tight regulation are instilled on immune responses. PMID- 17124514 TI - Progress and obstacles in the development of an AIDS vaccine. AB - Recent experimental observations suggest approaches to immunization that might finally result in at least a partially effective vaccine against infection with HIV-1. In particular, advances in our understanding of the contribution of vaccine-elicited cellular immunity to protecting memory CD4(+) T cells from virus mediated destruction provide rational strategies for the development of this vaccine. This is therefore an ideal time to review our current understanding of HIV-1 and its control by the immune system, as well as the remaining problems that must be solved to facilitate the development of an effective vaccine against AIDS. PMID- 17124515 TI - Perforin-mediated target-cell death and immune homeostasis. AB - The granule exocytosis pathway of cytotoxic lymphocytes is crucial for immune surveillance and homeostasis. The trafficking of granule components, including the membrane-disruptive protein perforin, to the immunological synapse leads to the delivery of granule proteases (granzymes) into the target cell and its destruction through apoptosis. Several independent molecular abnormalities associated with defects of either granule trafficking or perforin function can cause cytotoxic lymphocyte dysfunction. In humans, inherited perforin mutations result in severe immune dysregulation that manifests as familial haemophagocytic lymphohistiocytosis. This Review describes recent progress in defining the structure, function, biochemistry and cell biology of perforin. PMID- 17124516 TI - Living unrelated kidney donor transplantation in Iran. PMID- 17124517 TI - Case studies: more than simple anecdotes. PMID- 17124518 TI - Renal dysfunction in leptospirosis: a view from the tropics. PMID- 17124519 TI - Do statins have a beneficial effect on the kidney? PMID- 17124520 TI - Timing of sudden death relative to the hemodialysis procedure. PMID- 17124521 TI - Is cool dialysis an effective and well-tolerated means of reducing the frequency of intradialytic hypotension? PMID- 17124522 TI - Maintenance treatment of proliferative lupus nephritis can be discontinued after remission in some patients. PMID- 17124523 TI - Does the Adrogue-Madias formula accurately predict serum sodium levels in patients with dysnatremias? PMID- 17124524 TI - Bisphosphonates can improve bone mineral density in renal transplant recipients. PMID- 17124525 TI - Depression in end-stage renal disease hemodialysis patients. AB - Depression has been identified as a complicating comorbid diagnosis in a variety of medical conditions, including end-stage renal disease (ESRD). Despite this, the psychological health of hemodialysis patients is understudied. The purpose of this paper is to review the research and issues involved in the assessment of depression and its sequelae in ESRD. Accurate estimation of the prevalence of depression in the ESRD population has been difficult due to the use of different definitions of depression and varied assessment techniques, the overlap of depressive symptomatology with symptoms of uremia, and the confounding effects of medications. We suggest that depressive affect is a more important construct to study than diagnosis of depression syndromes per se in patients with chronic kidney disease. The Beck Depression Inventory is a reasonable measure of depressive affect in the ESRD population, if a higher than usual cutoff score is used or if its somatic components are omitted. Several pathways link depression and ESRD, and are probably bidirectional. As such, treatment of depressive affect could impact medical as well as psychological outcomes. The need for treatment intervention trials is great. Limited evidence regarding the safety and efficacy of treatment of hemodialysis patients with selective serotonin reuptake inhibitors is available, and cognitive behavioral therapy holds promise as an intervention for depression in this complex medical population. PMID- 17124526 TI - Drug insight: maintenance immunosuppression in kidney transplant recipients. AB - Kidney transplantation is the treatment of choice for patients with end-stage renal disease, in part because of ongoing efforts towards improving immunosuppressive strategies. Although calcineurin inhibitors remain the mainstay of immunosuppression in kidney transplant recipients, within this class of drug there has been a shift from use of ciclosporin to use of tacrolimus. Mycophenolate mofetil and mycophenolate sodium are now the antimetabolites of choice. A new class of drugs (inhibitors of mammalian target of rapamycin) that includes sirolimus is being increasingly used in stable kidney transplant recipients. New data, however, indicate that a more cautious approach to the use of this drug is warranted. Many transplant centers are now using steroid avoidance, minimization and withdrawal protocols. The impact of these different drugs and therapeutic strategies on outcomes has to be weighed against their immunosuppressive benefit. As more and more community-based nephrologists and primary care physicians are becoming involved in the care of stable kidney transplant recipients, it is important for these clinicians to familiarize themselves with novel immunosuppressive drugs and their pharmacokinetic properties. PMID- 17124527 TI - Mechanisms of disease: in utero programming in the pathogenesis of hypertension. AB - Nutritional and other environmental cues during development can permanently alter the structure, homeostatic systems, and functions of the body. This phenomenon has been referred to as 'programming'. Epidemiological and animal studies show that programmed effects operate within the normal range of growth and development, and influence the risk of chronic disease in adult life. We review the evidence that these effects include reduced nephron number and compensatory adaptations, which might lead to hypertension, and perhaps accelerate the decline in renal function that accompanies aging. These processes might be exacerbated by programmed changes in vascular structure and function, and alterations in endocrine and metabolic homeostasis. Programmed effects might be initiated as early as the periconceptual phase of development, and could involve epigenetic changes in gene expression or altered stem cell allocation. Better understanding of these processes could lead to the development of novel diagnostic and preventive measures, and to early detection of at-risk individuals. By monitoring blood pressure, weight, and renal function in children, it might be possible to reduce the risk of cardiovascular and renal disease in later life. PMID- 17124528 TI - Interpretation of serologic tests in an HIV-infected patient with kidney disease. AB - BACKGROUND: A 32-year-old African American man with HIV infection presented with hemoptysis, shortness of breath and renal insufficiency. Serologic testing revealed the presence of anti-glomerular basement membrane antibodies and equivocal levels of anti-myeloperoxidase antibodies. INVESTIGATIONS: Physical examination, urine and blood analysis, kidney ultrasound, chest radiograph, sputum cultures, bronchoscopy and renal biopsy. DIAGNOSIS: Reactivation of tuberculosis infection, immune complex glomerulonephritis, and 'false-positive' anti-glomerular basement membrane and anti-myeloperoxidase antibodies. MANAGEMENT: Directly observed therapy with four-drug anti-tuberculosis therapy and conservative management of chronic kidney disease. PMID- 17124529 TI - Safety and reactogenicity of an MSP-1 malaria vaccine candidate: a randomized phase Ib dose-escalation trial in Kenyan children. AB - OBJECTIVE: Our aim was to evaluate the safety, reactogenicity, and immunogenicity of an investigational malaria vaccine. DESIGN: This was an age-stratified phase Ib, double-blind, randomized, controlled, dose-escalation trial. Children were recruited into one of three cohorts (dosage groups) and randomized in 2:1 fashion to receive either the test product or a comparator. SETTING: The study was conducted in a rural population in Kombewa Division, western Kenya. PARTICIPANTS: Subjects were 135 children, aged 12-47 mo. INTERVENTIONS: Subjects received 10, 25, or 50 microg of falciparum malaria protein 1 (FMP1) formulated in 100, 250, and 500 microL, respectively, of AS02A, or they received a comparator (Imovax (rabies vaccine). OUTCOME MEASURES: We performed safety and reactogenicity parameters and assessment of adverse events during solicited (7 d) and unsolicited (30 d) periods after each vaccination. Serious adverse events were monitored for 6 mo after the last vaccination. RESULTS: Both vaccines were safe and well tolerated. FMP1/AS02A recipients experienced significantly more pain and injection-site swelling with a dose-effect relationship. Systemic reactogenicity was low at all dose levels. Hemoglobin levels remained stable and similar across arms. Baseline geometric mean titers were comparable in all groups. Anti-FMP1 antibody titers increased in a dose-dependent manner in subjects receiving FMP1/AS02A; no increase in anti-FMP1 titers occurred in subjects who received the comparator. By study end, subjects who received either 25 or 50 microg of FMP1 had similar antibody levels, which remained significantly higher than that of those who received the comparator or 10 microg of FMP1. A longitudinal mixed effects model showed a statistically significant effect of dosage level on immune response (F(3,1047) = 10.78, or F(3, 995) = 11.22, p < 0.001); however, the comparison of 25 microg and 50 microg recipients indicated no significant difference (F(1,1047) = 0.05; p = 0.82). CONCLUSIONS: The FMP1/AS02A vaccine was safe and immunogenic in malaria-exposed 12- to 47-mo-old children and the magnitude of immune response of the 25 and 50 microg doses was superior to that of the 10 microg dose. PMID- 17124530 TI - Safety and allele-specific immunogenicity of a malaria vaccine in Malian adults: results of a phase I randomized trial. AB - OBJECTIVES: The objectives were to evaluate the safety, reactogenicity, and allele-specific immunogenicity of the blood-stage malaria vaccine FMP1/AS02A in adults exposed to seasonal malaria and the impact of natural infection on vaccine induced antibody levels. DESIGN: We conducted a randomized, double-blind, controlled phase I clinical trial. SETTING: Bandiagara, Mali, West Africa, is a rural town with intense seasonal transmission of Plasmodium falciparum malaria. PARTICIPANTS: Forty healthy, malaria-experienced Malian adults aged 18-55 y were enrolled. INTERVENTIONS: The FMP1/AS02A malaria vaccine is a 42-kDa recombinant protein based on the carboxy-terminal end of merozoite surface protein-1 (MSP 1(42)) from the 3D7 clone of P. falciparum, adjuvanted with AS02A. The control vaccine was a killed rabies virus vaccine (Imovax). Participants were randomized to receive either FMP1/AS02A or rabies vaccine at 0, 1, and 2 mo and were followed for 1 y. OUTCOME MEASURES: Solicited and unsolicited adverse events and allele-specific antibody responses to recombinant MSP-1(42) and its subunits derived from P. falciparum strains homologous and heterologous to the 3D7 vaccine strain were measured. RESULTS: Transient local pain and swelling were more common in the malaria vaccine group than in the control group (11/20 versus 3/20 and 10/20 versus 6/20, respectively). MSP-1(42) antibody levels rose during the malaria transmission season in the control group, but were significantly higher in malaria vaccine recipients after the second immunization and remained higher after the third immunization relative both to baseline and to the control group. Immunization with the malaria vaccine was followed by significant increases in antibodies recognizing three diverse MSP-1(42) alleles and their subunits. CONCLUSIONS: FMP1/AS02A was well tolerated and highly immunogenic in adults exposed to intense seasonal malaria transmission and elicited immune responses to genetically diverse parasite clones. Anti-MSP-1(42) antibody levels followed a seasonal pattern that was significantly augmented and prolonged by the malaria vaccine. PMID- 17124531 TI - Proteasome-mediated degradation of IkappaBalpha and processing of p105 in Crohn disease and ulcerative colitis. AB - Enhanced NF-kappaB activity is involved in the pathology of both forms of inflammatory bowel disease (IBD), Crohn disease (CD) and ulcerative colitis (UC). Here we analyzed the mechanism of proteasome-mediated NF-kappaB activation in CD and UC. Our studies demonstrate that the subunit composition and the proteolytic function of proteasomes differ between UC and CD. High expression of the immunoproteasome subunits beta1i and beta2i is characteristic of the inflamed mucosa of CD. In line with this, we found enhanced processing of NF-kappaB precursor p105 and degradation of inhibitor of NF-kappaB, IkappaBalpha, by immunoproteasomes isolated from the mucosa of CD patients. In comparison with healthy controls and CD patients, UC patients exhibited an intermediate phenotype regarding the proteasome-mediated processing/degradation of NF-kappaB components. Finally, increased expression of the NF-kappaB family member c-Rel in the inflamed mucosa of CD patients suggests that p50/c-Rel is important for IFN-gamma mediated induction of immunoproteasomes via IL-12-driven Th1 responses. These findings suggest that distinct proteasome subunits influence the intensity of NF kappaB-mediated inflammation in IBD patients. PMID- 17124532 TI - Ca2+/calmodulin-dependent protein kinase II regulates cardiac Na+ channels. AB - In heart failure (HF), Ca(2+)/calmodulin kinase II (CaMKII) expression is increased. Altered Na(+) channel gating is linked to and may promote ventricular tachyarrhythmias (VTs) in HF. Calmodulin regulates Na(+) channel gating, in part perhaps via CaMKII. We investigated effects of adenovirus-mediated (acute) and Tg (chronic) overexpression of cytosolic CaMKIIdelta(C) on Na(+) current (I(Na)) in rabbit and mouse ventricular myocytes, respectively (in whole-cell patch clamp). Both acute and chronic CaMKIIdelta(C) overexpression shifted voltage dependence of Na(+) channel availability by -6 mV (P < 0.05), and the shift was Ca(2+) dependent. CaMKII also enhanced intermediate inactivation and slowed recovery from inactivation (prevented by CaMKII inhibitors autocamtide 2-related inhibitory peptide [AIP] or KN93). CaMKIIdelta(C) markedly increased persistent (late) inward I(Na) and intracellular Na(+) concentration (as measured by the Na(+) indicator sodium-binding benzofuran isophthalate [SBFI]), which was prevented by CaMKII inhibition in the case of acute CaMKIIdelta(C) overexpression. CaMKII coimmunoprecipitates with and phosphorylates Na(+) channels. In vivo, transgenic CaMKIIdelta(C) overexpression prolonged QRS duration and repolarization (QT intervals), decreased effective refractory periods, and increased the propensity to develop VT. We conclude that CaMKII associates with and phosphorylates cardiac Na(+) channels. This alters I(Na) gating to reduce availability at high heart rate, while enhancing late I(Na) (which could prolong action potential duration). In mice, enhanced CaMKIIdelta(C) activity predisposed to VT. Thus, CaMKII-dependent regulation of Na(+) channel function may contribute to arrhythmogenesis in HF. PMID- 17124533 TI - The uncertainty of a result from a linear calibration. AB - The standard error of a result obtained from a straight line calibration is given by a well known ISO-endorsed expression. Its derivation and use are explained and the approach is extended for any function that is linear in the coefficients, with an example of a weighted quadratic calibration in ICPAES. When calculating the standard error of an estimate, if QC data is available it is recommended to use the repeatability of the instrumental response, rather than the standard error of the regression, in the equation. PMID- 17124534 TI - Highly sensitive electrochemical detection of trace liquid peroxide explosives at a Prussian-blue 'artificial-peroxidase' modified electrode. AB - A highly sensitive electrochemical assay of the peroxide-based explosives triacetone triperoxide (TATP) and hexamethylene triperoxide diamine (HMTD) at a Prussian-blue (PB) modified electrode is reported. The method involves photochemical degradation of the peroxide explosives and a low potential (0.0 V) electrocatalytic amperometric sensing of the generated hydrogen peroxide at the PB transducer and offers nanomolar detection limits following a short (15 s) irradiation times. The electrochemical sensing protocol should facilitate rapid field screening of peroxide explosives. PMID- 17124535 TI - K+-selective nanospheres: maximising response range and minimising response time. AB - Cross-linked K(+) ion-selective copolymer nanospheres have been prepared by free radical photo-initiated polymerization of n-butyl acrylate (nBA) with hexanedioldiacrylate (HDDA). Nanospheres (<200 nm) containing H(+) chromoionophore (ETH 5294) and lipophilic salt (KTClPB) for H(+)-sensors, or ETH 5294, a K(+)-selective ionophore (valinomycin) and anionic sites for K(+)-sensors were compared, and the effect of varying the normalised concentrations for beta (R(T)(-)/L(T)) and gamma (C(m)(T)/L(T)) was studied. Experimental data were fitted to theoretical curves for the dynamic response range, based on the effect of changes in the concentration of these lipophilic sensing components incorporated into the spheres, and conditions identified for maximising the response range. A complex valinomycin-K(+) formation constant, log K(IL) = 13.13 +/- 2.22, was obtained in the nBA matrix, and from the calibration curves the apparent acid-dissociation equilibrium constant (pK(a) = 12.92 +/- 0.03) was extracted for the H(+)-sensing system, and the equilibrium exchange constant (pK(exch) = 6.16 +/- 0.03, at pH 7) calculated for the K(+)-sensing nanospheres. A basis for establishing optimum performance was identified, whereby response range and response time were balanced with maximum fluorescence yield. Parameters for achieving nanospheres with a response time <5 minutes, covering 2-3 orders of magnitude change in activity were identified, demanding nanospheres with radius <300 nm and beta(crit) approximately 0.6. An RSD(%) approximately 3% was obtained in a study of the reproducibility of the response of the proposed nanospheres, and selectivity was also evaluated for a K(+)-selective nanosensor using several cations as interfering agents. In most cases, the fluorescent emission spectra showed no response to the cations tested, confirming the selectivity of nanospheres to potassium ion. The nanosensors were satisfactorily applied to the determination of K(+) in samples mimicking physiological conditions. PMID- 17124536 TI - The advantage of using carbon nanotubes compared with edge plane pyrolytic graphite as an electrode material for oxidase-based biosensors. AB - Carbon nanotubes (CNTs) are promising materials for use in amperometric biosensors. The defect sites at their ends, and on their sidewalls, are considered to be edge plane-like defects and show high electrocatalytic activity toward several biological molecules. However, electrocatalytic activity toward H(2)O(2) has not been compared among bamboo-structured CNTs (BCNTs), which have many defect sites; hollow-structured CNTs (HCNTs), which have few defect sites; edge plane pyrolytic graphite (EPG); and traditional glassy carbon (GC). The advantages of using CNTs in electrodes for biosensors are still equivocal. To confirm the utility of CNTs, we analyzed the electrochemical performance of these four carbon electrodes. The slope of the calibration curve for H(2)O(2) at potentials of both +0.6 V and -0.1 V obtained with a BCNT paste electrode (BCNTPE) was more than 10 times greater than the slopes obtained with an HCNT paste electrode and a GC electrode, reflecting the BCNT's larger number of defect sites. Although the slope with the EPG electrode (EPGE) was about 40 times greater than that with BCNTPE at +0.6 V, the slopes with these two carbon electrodes were nearly equivalent at -0.1 V. EPGE demonstrated excessive electrochemical activity, detecting currents on the basis of consumption of oxygen and oxidation of ascorbic acid, even at -0.1 V. In contrast, BCNTPE could dominantly detect a cathodic current for H(2)O(2) at -0.1 V, even when interfering molecules were added. BCNTPE possesses appropriate electrochemical activity and is an effective electrode materials for developing interference-free oxidase-based biosensors operated by the application of an appropriate potential. PMID- 17124537 TI - Chemical analysis in the submillimetre spectral region with a compact solid state system. AB - A new analytical system that uses the rotational signatures of gas phase molecules is described and demonstrated. It uses a solid state source to probe molecular systems in the millimetre and submillimetre wave range, the only region of the electromagnetic spectrum not yet used extensively for analytical purposes. It employs the FAst Scan Submillimetre Spectroscopy Technique (FASSST), which leads to an especially simple system architecture. Among the attributes of the system are generality, sensitivity, 'absolute' specificity, small size, simplicity, and the potential for very low cost. Applications to problems of analytical interest are also discussed. PMID- 17124538 TI - Excitation-emission matrix fluorescence spectroscopy in conjunction with multiway analysis for PAH detection in complex matrices. AB - A field portable, single exposure excitation-emission matrix (EEM) fluorometer has been constructed and used in conjunction with parallel factor analysis (PARAFAC) to determine the sub part per billion (ppb) concentrations of several aqueous polycyclic aromatic hydrocarbons (PAHs), such as benzo(k)fluoranthene and benzo(a)pyrene, in various matrices including aqueous motor oil extract and asphalt leachate. Multiway methods like PARAFAC are essential to resolve the analyte signature from the ubiquitous background in environmental samples. With multiway data and PARAFAC analysis it is shown that reliable concentration determinations can be achieved with minimal standards in spite of the large convoluting fluorescence background signal. Thus, rapid fieldable EEM analyses may prove to be a good screening method for tracking pollutants and prioritizing sampling and analysis by more complete but time consuming and labor intensive EPA methods. PMID- 17124539 TI - Lectin affinity chromatography using porous polymer monolith assisted nanoelectrospray MS/MS. AB - An affinity porous polymer monolith is utilized as a nanoelectrospray emitter as well as an online affinity capture column for the preconcentration of glycans. Porous polymer monolith (PPM) assisted electrospray provides a facile methodology for coupling microfluidics to mass spectrometry that is sheathless and with zero dead volume. Affinity PPM was photopolymerized using glycidyl methacrylate/ethylene dimethacrylate utilizing different porogenic solvents based on aliphatic alcohols to provide PPMs with a variety of pore sizes. The use of longer alkyl chain alcohols decreased the pore size of the formed PPM as indicated by the higher flow back pressure generated. The effect of the pore size on the stability of the electrospray was tested showing higher stability of the TIC with lower pore size. A lectin, namely Concanavaline A, was immobilized on glycidyl methacrylate/ethylene dimethacrylate using the Schiff base method to provide an affinity monolith for high mannose glycans. The amount of the lectin immobilized was studied as a function of the porogenic solvent used in the polymerization. The glycopeptides of the glycoprotein Ribonuclease B was preconcentrated on the affinity PPM sprayer and detected by tandem MS. PMID- 17124540 TI - Redox magnetohydrodynamic enhancement of stripping voltammetry: toward portable analysis using disposable electrodes, permanent magnets, and small volumes. AB - The use of redox magnetohydrodynamics (MHD) to enhance the anodic stripping voltammetry (ASV) response of heavy metals has been investigated, with respect to achieving portability: disposable electrodes consisting of screen-printed carbon (SPC) on a low temperature co-fired ceramic (LTCC) substrate, small volumes, and permanent magnets. The analytes tested (Cd(2+), Cu(2+), and Pb(2+)) were codeposited on SPC with Hg(2+) to form a Hg thin film electrode. High concentrations of Fe(3+) were used to produce a high cathodic current which generates a significant Lorentz force in the presence of a magnetic field. This Lorentz force induces solution convection during the deposition step, enhancing the mass transport of analytes to the electrode and increasing their preconcentrated quantity in the mercury thin film. Therefore, larger ASV peaks and improved sensitivities are obtained, compared to analyses performed without a magnet. The effects on ASV signal of varying Hg(2+) concentration (0.10 and 1.0 mM), deposition time (10-600 s), and electrode surface roughness were investigated. In addition, analyses were performed using a real lake water matrix. By using the disposable LTCC-SPC working electrodes in small volumes (150 microL) and with small permanent magnets (0.78 T), peak areas were increased by 75% when compared to the signal obtained in the absence of a magnetic field. A limit of detection of 25 nM for Cd(2+) was observed with only a 1 min preconcentration time. PMID- 17124541 TI - Separation of samarium and neodymium: a prerequisite for getting signals from nuclear synthesis. AB - (146)Sm (T(1/2) = 10(8) y) is a long-lived radionuclide which has been produced in significant amounts during burning in a supernova (SN). Detection of this SN produced long-lived radionuclide on Earth may be helpful for getting information on nuclear synthesis at the time of our solar system's formation. Only accelerator mass spectrometry (AMS) can determine such minute traces of (146)Sm still expected in the Earth's crust. However, the villain of (146)Sm measurement through AMS is its naturally occurring stable isobar (146)Nd which is a million times more abundant than the trace amount of (146)Sm. Therefore an efficient method for the separation of samarium and neodymium is required to measure (146)Sm through AMS. A simple liquid-liquid extraction (LLX) based method for separation of samarium and neodymium has been developed using radiometric simulation. Di-(2-ethylhexyl)phosphoric acid (HDEHP) has been used as the organic reagent. A very high separation factor ( approximately 10(6)) can be achieved when a solution containing samarium and neodymium is reduced by hydroxylamine hydrochloride followed by extraction with 0.1% HDEHP diluted in cyclohexane from 0.025 M HCl solution. PMID- 17124542 TI - Comparison of spectral counting and metabolic stable isotope labeling for use with quantitative microbial proteomics. AB - Spectral counting, a promising method for quantifying relative changes in protein abundance in mass spectrometry-based proteomic analysis, was compared to metabolic stable isotope labeling using (15)N/(14)N "heavy/light" peptide pairs. The data were drawn primarily from a Methanococcus maripaludis experiment comparing a wild-type strain with a mutant deficient in a key enzyme relevant to energy metabolism. The dataset contained both proteome and transcriptome measurements. The normalization technique used previously for the isotopic measurements was inappropriate for spectral counting, but a simple adjustment for sampling frequency was sufficient for normalization. This adjustment was satisfactory both for M. maripaludis, an organism that showed relatively little expression change between the wild-type and mutant strains, and Porphyromonas gingivalis, an intracellular pathogen that has demonstrated widespread changes between intracellular and extracellular conditions. Spectral counting showed lower overall sensitivity defined in terms of detecting a two-fold change in protein expression, and in order to achieve the same level of quantitative proteome coverage as the stable isotope method, it would have required approximately doubling the number of mass spectra collected. PMID- 17124543 TI - Voltammetric analysis of europium at screen-printed electrodes modified with salicylamide self-assembled on mesoporous silica. AB - Mercury-free sensors for europium (Eu(3+)) assay based on the chemical modification of screen-printed carbon electrodes (SPCEs) with self-assembled salicylamide on mesoporous silica (Sal-SAMMS) have been developed. The preconcentration of Eu(3+) at SAMMS-based sensors utilizes the binding affinity of the salicylamide and Eu(3+), accomplished at open circuit potential without electrolyte and solution de-gassing. Optimal Eu detection was obtained after 3-5 min preconcentration in Eu solution (pH 2-6), electrolysis at -0.9 V for 60 s in a new medium (0.1-0.2 M NH(4)Cl, pH 3.5), followed by a square-wave voltammetric detection of Eu in the same electrolyte. Attributed to the strong covalent bonding of the functional groups on mesoporous silica and silane cross-linking, the SAMMS-modified SPCEs with a built-in 3-electrode system can be re-used for tens of measurements with minimal degradation, enabling the establishment of the calibration curve and lowering the costs. A linear calibration curve was found in the range of 75 to at least 500 ppb Eu(3+) after 5 min preconcentration. The experimental detection limit was 10 ppb after 10 min preconcentration, which can be improved with increased preconcentration time. Reproducibility (% RSD) of 100 ppb Eu(2+) was 10% for a single sensor and 10% for 5 sensors, which can be improved through the precision of sensor manufacturing, in which SAMMS modification can be made in-situ. PMID- 17124544 TI - Differences in the degree of inhibition of NDP reductase by chemical inactivation and by the thermosensitive mutation nrdA101 in Escherichia coli suggest an effect on chromosome segregation. AB - NDP reductase activity can be inhibited either by treatment with hydroxyurea or by incubation of an nrdA (ts) mutant strain at the non-permissive temperature. Both methods inhibit replication, but experiments on these two types of inhibition yielded very different results. The chemical treatment immediately inhibited DNA synthesis but did not affect the cell and nucleoid appearance, while the incubation of an nrdA101 mutant strain at the non-permissive temperature inhibited DNA synthesis after more than 50 min, and resulted in aberrant chromosome segregation, long filaments, and a high frequency of anucleate cells. These phenotypes are not induced by SOS. In view of these results, we suggest there is an indirect relationship between NDP reductase and the chromosome segregation machinery through the maintenance of the proposed replication hyperstructure. PMID- 17124545 TI - Spectroscopic studies of D-alpha-tocopherol concentration-induced transformation in egg phosphatidylcholine vesicles. AB - The effects of embedding up to 60 mol% of alpha-tocopherol (alpha-Toc) on the morphology and structure of the egg phosphatidylcholine (PC) membrane were studied using spectroscopic techniques. The resulting vesicles were subjected to turbidometric and dynamic light scattering measurements to evaluate their size distribution. The alpha-Toc intrinsic fluorescence and its quenching was used to estimate the tocopherol position in the membrane. Optical microscopy was used to visualize morphological changes in the vesicles during the inclusion of tocopherol into the 2 mg/ml PC membrane. The incorporation of up to 15 mol% of tocopherol molecules into PC vesicles is accompanied by a linear increase in the fluorescence intensity and the simultaneous formation of larger, multilamellar vesicles. Increasing the tocopherol concentration above 20 mol% induced structural and morphological changes leading to the disappearance of micrometer sized vesicles and the formation of small unilamellar vesicles of size ranging from 30 to 120 nm, mixed micelles and non-lamellar structures. PMID- 17124546 TI - A proteomic analysis of the effect of MAPK pathway activation on L-glutamate induced neuronal cell death. AB - Oxidative stress has been implicated in the pathogenesis of neuronal degenerative diseases. It is also widely known that oxidative stress induces mitogen-activated protein kinase (MAPK) signaling cascades. In this study, we used proteomic analysis to investigate the role of the MAPK pathway in oxidative stress-induced neuronal cell death. The results demonstrated that several proteins, including eukaryotic translation elongation factor 2 (eEF2) and enolase I, showed a differential expression pattern during the neuronal cell death process, and this was MAPK pathway dependent. Several chaperone and cytoskeletal proteins including heat shock protein 70, calreticulin, vimentin, prolyl 4-hydroxylase beta polypeptide, and transgelin 2 were up-or down-regulated, despite their expressions not depending on the MAPK pathway. These findings strongly suggest that the expressions of proteins which play protective roles are independent of the MAPK pathway. On the other hand, eEF2 and enolase I may be the downstream targets of the MAPK pathway. PMID- 17124547 TI - Tagging target genes of the MAT1-2-1 transcription factor in Fusarium verticillioides (Gibberella fujikuroi MP-A). AB - Mating type in filamentous ascomycetes is controlled by idiomorphic alleles, named MAT1-1 and MAT1-2, which contain 1-3 genes. Of these genes MAT1-1-1 and MAT1-2-1 encode putative transcription factors and are thus considered to be the major regulators of sexual communication and mating. Fungi with no known sexual stage may also have fully functional mating type genes and therefore it was plausible to hypothesize that the MAT products may also regulate other types of genes not involved directly in the mating process. To identify putative target genes of these transcription factors in Fusarium verticillioides, DeltaMAT1-2-1 knock out mutants were produced and transcript profiles of mutant and wild type were compared by means of differential cDNA hybridization. Clones, either up- or down-regulated in the DeltaMAT1-2-1 mutant were sequenced and a total of 248 sequences were blasted against the NCBI database as well as the Gibberella zeae and Gibberella moniliformis genomes. Fifty-five percent of the clones were down regulated in the mutant, indicating that the MAT1-2-1 product positively affected these tagged sequences. On the other hand, 45% were found to be up-regulated in the mutant, suggesting that the MAT1-2-1 product also exerted a negative regulatory function on this set of genes. Sequences involved in protein synthesis and metabolism occurred more frequently among the clones up-regulated in the mutant, whereas genes belonging to cell signalling and communication were especially frequently tagged among the sequences down-regulated in the mutant. PMID- 17124548 TI - Dietary intake of B-vitamins in mothers born a child with a congenital heart defect. AB - BACKGROUND: Periconceptional use of multivitamins reduces the risk of a child with a congenital heart defect (CHD). Data on the impact of maternal diet, however, are lacking. AIM OF THE STUDY: We investigated the association between the maternal dietary intake of B-vitamins and having a child with a CHD. METHODS: A case-control study was performed in 192 mothers of a child with a CHD and 216 mothers of a healthy child. Mothers filled out food frequency questionnaires covering the current dietary intake, and general questionnaires at 17 months after the index-pregnancy. Maternal blood samples were taken to determine B vitamin and plasma total homocysteine (tHcy) concentrations as nutritional biomarkers. Pregnant and lactating mothers and those with another diet compared with the preconceptional period were excluded for analysis. Case-mothers and controls were compared using the Mann-Whitney U test and logistic regression. RESULTS: The dietary intake of macronutrients and B-vitamins was comparable between both groups, but all mothers had a substantially lower median folate intake (cases 161 microg, controls 175 microg) than the Dutch recommended dietary allowance of 300 microg. Within the case-group, the intake of proteins and vitamin B(6) and the concentrations of serum vitamin B(12) and folate were significantly lower in hyperhomocysteinemics (tHcy > or = 14.5 micromol/l) than in normohomocysteinemics. The maternal educational level was positively associated with B-vitamin intake, except for vitamin B(12) in controls. Low educated case-mothers showed a significantly lower median vitamin B(12) intake than controls (2.8 microg and 3.8 microg, P = 0.01). The CHD risk doubled if vitamin B(12) intake in these mothers reduced by 50% (OR 2.0; 95% CI: 1.1-3.5). CONCLUSIONS: A diet low in vitamin B(12) is associated with an increased risk of a child with a CHD, especially in low educated women. A disbalance in the maternal intake of proteins and low folate intake may play a role as well, but needs further investigation. As hyperhomocysteinemia is a strong risk factor for adult cardiovascular disease, these data may imply that the hyperhomocysteinemic mothers and their children should be targeted for nutritional interventions. PMID- 17124550 TI - The use of mesh and collagen in incisional hernias. PMID- 17124551 TI - Measuring quality of life in rheumatic conditions. AB - Musculoskeletal disorders often have associated pain, functional impairment and work disability, and, not surprisingly, are the most common reasons for utilizing healthcare resources. Rheumatoid arthritis (RA) and fibromyalgia (FM) are causes of musculoskeletal pain and disability. Research indicates that there is a widespread impact of RA and FM on physical, psychological and social factors in affected individuals, and thus, outcome measures that encompass multiple aspects of quality of life are needed. Generic measures of quality of life identify associations between physical conditions and mental health and highlight the need to address psychological functioning to ultimately improve the individuals' quality of life. PMID- 17124552 TI - Assessment of osteoclast number and function: application in the development of new and improved treatment modalities for bone diseases. AB - Numerous experimental and clinical observations suggest that overall changes in bone resorption during menopause or treatment with hormone replacement therapy (HRT) are combined effects of changes in osteoclast number and function. Moreover, due to a coupling between osteoclastic bone resorption and osteoblastic bone formation, pronounced alteration of osteoclast number will eventually lead to alteration of osteoblastic bone formation. Fragments of type I collagen, such as the C- and N-terminal telopeptides of collagen type I (CTX and NTX, respectively), are generated during bone resorption and hence can be used as surrogate markers of osteoclast function. Circulating levels of different enzymes in the serum, such as TRAP 5b and cathepsin K are proportional to the number of osteoclasts, and hence can be used as surrogate markers of osteoclast number. Since antiresorptive effects can be obtained in different ways, we felt it was timely to discuss the different scenarios, highlight differences specific to different pharmacological interventions with different mechanisms of action, and discuss how these bone markers can assist us in a deeper analysis of the pharmacodynamics and safety profile of existing and upcoming drug candidates. PMID- 17124553 TI - Gender differences in the ratio between humerus width and length are established prior to puberty. AB - SUMMARY: On a sample of 1,317 children aged 9.9 years we developed a novel method of measuring humeral dimensions from total body dual-energy X-ray absorptiometry (DXA) scans and showed that gender differences in the ratio between humeral width and length are established prior to puberty. INTRODUCTION: It is recognised that long bone cross-sectional area is greater in males compared to females, which is thought to reflect more rapid periosteal bone growth in boys. However, it is currently unclear whether these findings reflect gender differences in bone size or shape. In the present study, we investigated whether gender differences exist in the balance between longitudinal and periosteal long bone growth in children, leading to gender differences in bone shape, based on a novel method for evaluating shape of the humerus. We also examined whether these differences are established prior to puberty. METHODS: Length, area and width of the humerus were estimated from total body DXA scans in 1,317 children aged 9.9 +/- 0.33 years, who had participated in a nested case-control study of fractures within the Avon Longitudinal Study of Parents and Children (ALSPAC) (a geographically based birth cohort based in South West England). No differences were observed with respect to parameters of humeral geometry according to fracture history, and so both groups were pooled for further analysis. Aspect ratio (AR) of the humerus was calculated as humeral width divided by length. Total body height and weight were measured at the same time as the DXA scan. Puberty was assessed using self-completion questionnaires. RESULTS: Humeral width and length were positively associated with age and height in boys and girls combined (P<0.001), and with Tanner stage in girls (P<0.002). In contrast, age, height and Tanner stage were not related to humeral AR. We then examined gender differences in humeral shape according to pubertal stage. In prepubertal children (i.e. Tanner stage 1), humeral length was similar in boys and girls, but width (1.92 vs 1.88 cm, P<0.001) and area (47.7 vs 46.9 cm(2), P<0.001) were greater in boys, resulting in a greater AR (7.78 vs 7.53, P<0.001). Similar gender differences were observed in early pubertal children (i.e. Tanner stage 2). CONCLUSION: We conclude that the greater periosteal diameter of boys compared to girls reflects differences in the balance between longitudinal and periosteal bone growth. Interestingly, resulting gender differences in humeral AR are established in prepubertal children. PMID- 17124554 TI - A synthetic uracil derivative with antitumor activity through decreasing cyclin D1 and Cdk1, and increasing p21 and p27 in MCF-7 cells. AB - The anticarcinogenic potential of (RS)-1-(2,3-dihydro-5H-1,4-benzodioxepin-3 yl)uracil (DBDU), with the naturally occurring pyrimidine base uracil, is reported against the MCF-7 cancer cell line. The arrest in the G0/G1 and G2/M cell cycle phases was accounted for by decrease in the expression of the cyclin D1 and Cdk1 proteins, and increase in p21 and p27 proteins. Using a reverse transcription-polymerase chain reaction-based assay at a dose of 5 muM of DBDU cyclin D1 mRNA was decreased, suggesting that DBDU exerts its regulatory action on cyclin D1 at the level of transcription. DNA fragmentation was performed and demonstrated that apoptosis occurred in the tumor cell line treated with DBDU. The G0/G1 arrest is an irreversible process and the cells undergo apoptosis in a p53-independent manner. DBDU administered intravenously twice a week (50 mg/kg dose each time) induced neither toxicity nor death in mice for 5 weeks. PMID- 17124555 TI - Mid-region parathyroid hormone-related protein (PTHrP) binds chromatin of MDA MB231 breast cancer cells and isolated oligonucleotides "in vitro". AB - We have previously shown that PTHrP(38-94)-amide restrains growth and invasion "in vitro", causes striking toxicity and accelerates death of some breast cancer cell lines, the most responsive being MDA-MB231 whose tumorigenesis was also attenuated "in vivo". PTHrP(38-94)-amide contains the domain implicated in the nuclear import of PTHrP. Although the nucleus was identified as a destination for mid-region PTHrP, evidence for direct DNA-binding capability is lacking to date. Here, we examined the localization of PTHrP(38-94)-amide within MDA-MB231 cells and within metaphase spread preparations and characterized its DNA-binding properties, employing a combination of immunocytochemical, cytogenetic, "whole genome"/conventional PCR, EMSA and DNase footprinting techniques. The results obtained: (i) show that PTHrP(38-94)-amide gains access to the nuclear compartment of MDA-MB231 cell; (ii) demonstrate that PTHrP(38-94)-amide is a DNA binding peptide; and, (iii) represent the first data to date on the potential molecular targets in both cellular chromatin and isolated oligonucleotides "in vitro". PMID- 17124556 TI - Cloning and expression of heterologous genes in Rhodothermus marinus. AB - The construction of a shuttle cloning system for Rhodothermus marinus and Escherichia coli is described. It is based on the shuttle vector pRM3000, which contains a multiple cloning site as well as the shuttle marker trpB and TrpB(-) recipients of both species. The vector is stable and in 25 +/- 2 and 91 +/- 3 copies in R. marinus SB-1 and E. coli SDH-1, respectively. Three different R. marinus regulatory sequences of the dnaJ, dnaK and recA genes were integrated into pRM3000 to make the expression vectors pRM5100, pRM5200 and pRM5300, respectively. Genes encoding alpha- and beta-galactosidase (agaT and bgaT) from Thermus brockianus were cloned in R. marinus. Expression of both recombinant genes in R. marinus was demonstrated. The agaT gene was used as a reporter to study transcription from the expression vectors. Induced expression by ten- and twentyfold was observed from the dnaK and dnaJ regulatory sequences, respectively, after a temperature shift from 63 to 77 degrees C. This is the first report of cloning and expression of heterologous genes in R. marinus and the first study of promoter activity in the species. PMID- 17124557 TI - New insights in the treatment strategy for pulmonary arterial hypertension. AB - INTRODUCTION: Recent advances in our understanding of the pathophysiological and molecular mechanisms involved in pulmonary arterial hypertension have led to the development of novel and rational pharmacological therapies. In addition to conventional therapy (i.e., supplemental oxygen and calcium channel blockers), prostacyclin or endothelin receptor antagonists have been recommended as a first line therapy for pulmonary arterial hypertension. However, these treatments have potential limitations with regard to their long-term efficacy and improvement in survival. Furthermore, intravenous prostacyclin (epoprostenol) therapy, which is recommended by most experts for patients with New York Heart Association (NYHA) functional class IV, is complicated, uncomfortable for patients, and expensive because of the cumbersome administration system. Considering these circumstances, it is necessary to develop additional novel therapeutic approaches that target the various components of this multifactorial disease. CASE REPORT: In this short review, we present an overview of the current treatment options for pulmonary arterial hypertension and describe a case report with primary pulmonary hypertension. A male patient with NYHA functional class IV and showing no response to calcium channel blockers and prostacyclin exhibited significantly improved exercise tolerance and hemodynamics and long-term survival for more than 2.5 years after receiving an oral combination therapy of a phosphodiesterase type 5 inhibitor (sildenafil), phosphodiesterase type 3 inhibitor (pimobendan), and nicorandil. FUTURE PERSPECTIVE: We also discuss the background and plausible potential mechanisms involved in this case, as well as future perspectives in the treatment of pulmonary arterial hypertension. PMID- 17124559 TI - Examiner differences in the assessment of different malocclusions. AB - BACKGROUND: In the current discussion about ensuring treatment quality and reducing costs in the health sector, indication systems with which to determine the need for a treatment and the success of therapy are increasingly being used in orthodontics. These indication systems require the objective evaluation of malocclusions. Our objective was to determine the examiner reliability in the assessment of various malocclusions. MATERIALS AND METHODS: In 180 adults (64 male, 116 female, aged 20-49) from the population-based Study of Health in Pomerania (SHIP), malocclusions were recorded clinically and on models by calibrated examiners. An experienced orthodontist conducted the clinical examination. Another orthodontically-experienced examiner analyzed the models. To compare the model examiners, two examiners with varying degrees of orthodontic experience evaluated 60 of the 180 models as well (29 male, 31 female). One of the model examiners repeated the assessment of 60 models at a later time (intra individual comparison). RESULTS AND CONCLUSIONS: Reliability amongst the examiners depended on which malocclusions were judged: crowding and contact point displacement showed little agreement, while cross bite, edge-to-edge bite, deep bite and enlarged overjet revealed greater agreement. Comparison between the clinical examination and model analysis (kappa median 0.57) revealed the greatest differences between the examiners. Comparison of the three model examiners also showed differences. The contrast to the orthodontically least experienced examiner was greater (kappa median 0.61 and 0.62) than the divergence between the two orthodontically more experienced examiners (kappa median 0.70). The intra individual examiner comparison revealed the smallest differences (kappa median 0.82). PMID- 17124558 TI - Highly purified omega-3 fatty acids for secondary prevention of sudden cardiac death after myocardial infarction-aims and methods of the OMEGA-study. AB - INTRODUCTION: During the last decades a large body of data has been accumulated indicating omega-3 fatty acids to exert beneficial effects on the prognosis of patients with cardiovascular disease. Especially, omega-3 fatty acids are regarded to be effective in reducing the risk of sudden cardiac death after acute myocardial infarction. However, treatment of acute myocardial infarction and secondary prevention considerably have been improved within the past years including early revascularization by PCI, the routine use of beta-blockers, statins and ACE-inhibitors as well as cardiac rehabilitation for improving life style measures. To date, there exists no controlled randomized trial testing the prognostic effect of omega-3 fatty acids after acute myocardial infarction in a double blind regimen under the conditions of modern treatment of myocardial infarction. MATERIALS AND METHODS: The present study therefore evaluates the effect of highly purified omega-3 fatty acid ethylesters (omega-3-acid ethyl esters 90=Zodin) on the rate of sudden cardiac death within 1 year after acute myocardial infarction. Secondary endpoints are total mortality, non-fatal cardiovascular events, rhythm abnormalities in holter monitoring and depression score. RESULT AND CONCLUSION: The recruitment-period started in October 2003 and is expected to last until December 2006. The results of the study are therefore expected for the beginning of 2008, when all patients will have completed the 12 months follow up-period. PMID- 17124560 TI - Prevalence and development of KIG-relevant symptoms in primary school students from Frankfurt am Main. AB - OBJECTIVE: To determine the actual need for orthodontic treatment in the late mixed dentition according to the German KIG system (Kieferorthopadische Indikationsgruppen = Orthodontic Indication Groups). By comparing the findings with those of the early mixed dentition, we aimed to evaluate whether a change in the spectrum of malocclusions would occur. SUBJECTS AND METHODS: Dental check-ups took place in schools in Frankfurt am Main, during which the orthodontically relevant symptoms of 1251 schoolchildren (female 49.5%, male 50.5%) in grades 4 and 5 were recorded and compared with findings documented 4 years earlier in the same classes at the same schools. RESULTS: Of those orthodontic malocclusions in need of therapy and covered by the statutory health insurance, lateral crossbite and enlarged overjet (more than 6 mm) dominated by 9.2% and 8.7%, respectively, among the 9- to 11-year-old children. Of all the malocclusions recorded within the KIG-group showing a treatment need degree > or = 3, category D4 (overjet greater than 6 mm) at 17.4% was the most frequent, followed by K4 (unilateral crossbite) at 15.3%, and M4 (negative overjet up to -3 mm) at 14.9%. 41.4% of all the children examined presented a treatment indication according to statutory health insurance directives (KIG > or = 3). The 10% reduction in treatment cases financed by statutory health insurance has been achieved in any case [1], as mandated by health policy. CONCLUSIONS: Our comparison of results gathered from 2000 and 2004 showed that as the children grew older, the prevalence of already enlarged overjets increased, as did the frequency of deep bite. We observed fewer frontal open bites and crossbites in late mixed dentition. Treatment need according to current statutory health insurance directives was clearly higher in the late mixed dentition (41.4%) than in the early mixed dentition (8%). PMID- 17124561 TI - Magnetic forces on orthodontic wires in high field magnetic resonance imaging (MRI) at 3 tesla. AB - BACKGROUND: In a previous investigation we reported on magnetic forces in the static magnetic field of a 1.5 Tesla MRI system. The aim of the present investigation was to assess forces on orthodontic wires in a high field strength MRI system at 3 Tesla. MATERIALS AND METHODS: Thirty-two different orthodontic wires (21 archwires, eight ligature wires and three retainer wires) were investigated in a 3 Tesla high field strength MRI system (Intera, Philips Medical Systems, Best, The Netherlands). Translational forces were measured by the deflection angle test (ASTM F2052-02), and rotational forces assessed on a 5 point qualitative scale. RESULTS AND CONCLUSION: Translational forces ranged between 43.5 mN and 136.1 mN for retainer wires and between 0.6 mN (Noninium) and 208.4 mN (Orthos Stainless Steel) for steel archwires. Translational forces were up to 53.8 times as high as gravitational forces for retainer wires and up to 54.5 times as high for steel archwires, associated with marked rotational forces for the most part. Archwires manufactured from nickel-titanium, titanium molybdenum and cobalt-chromium and different ligature wires showed no or negligible forces in the magnetic field. Carefully ligated wires should not present a risk due to translational and rotational forces in the high field MRI system at 3 Tesla. PMID- 17124562 TI - Is the buccolingual crown diameter for transverse malocclusions? AB - OBJECTIVE: We investigated whether there is a correlation between the buccolingual crown diameter and cusp tip distance and if that can be considered a factor influencing the formation of a transversal malocclusion. MATERIALS AND METHODS: The buccolingual crown diameters and cusp tip distances of all premolars and first molars of 102 normal dentitions and 123 dentitions with a transverse malocclusion were measured and examined for significant differences. We also investigated whether there were differences in size between the genders and between left and right sides. RESULTS: General differences in buccolingual crown diameters ranged from 0.3 mm to 0.6 mm, and in cusp tip distance from 0.4 mm to 0.7 mm. Buccolingual crown width was generally greater in boys than girls, whereas we observed no statistically significant difference in cusp tip distances. Comparison of left and right sides revealed no significant difference. We were able to statistically prove the correlation between buccolingual tooth diameter and cusp tip distance. CONCLUSIONS: The results suggest that larger teeth with a greater cusp tip distance offer a more favorable prerequisite for the appearance of regular transversal occlusal relations than those with a smaller distance. The buccolingual crown width and its corresponding cusp tip distance seem important for initial contact with the antagonists, and may be regarded as a factor that determines whether or not a crossbite develops. PMID- 17124563 TI - The displaced maxillary canine--a retrospective study. AB - OBJECTIVE: The re-alignment of retained maxillary canines was studied in relation to the degree of displacement in panoramic radiographs, mesiodistal root deviations, length of treatment and side-effects. The aim was then to determine whether the decision between surgical removal and orthodontic re-alignment can be made on the basis of panoramic radiographs alone. MATERIALS AND METHODS: Forty seven panoramic radiographs with 59 displaced canines were used to determine their distance and inclination to the occlusal plane (similar to the method described by Dausch-Neumann [8]), the position of the crown tip, and the frequency of mesiodistal root deviations. The length of active treatment was recorded and analyzed statistically in relation to the degree of displacement and incidence of root deviation. Finally, pre- and post-therapeutic panoramic radiographs were compared to determine the incidence of root resorptions and marginal bone defects. RESULTS: The mean values for the angle of inclination and the distance of the maxillary canines from the occlusal plane were 57.4 degrees (+/- 14.3 degrees ) and 10.5 mm (+/- 3.8 mm), respectively. In 21 canines, the crown tips projected most frequently between the central and lateral incisors. Twelve canines presented root deviations. Treatment lasted an average of 1.9 years (+/- 0.7 years). No correlation was found between the treatment time and the canine's distance from the occlusal plane (r = 0.03), its angle of inclination (r = 0.06), position of the crown tip (r = 0.12), or root deviation (r = -0.07). Root resorptions were found in nine canines (15.3%) and marginal bone defects in 24 canines (40.7%). CONCLUSION: Fifty-nine retained maxillary canines, some showing extreme displacement, were re-aligned in an average of 1.9 years. There was no correlation between the degree of canine displacement in the panoramic radiographs or the incidence of mesiodistal root deviations and active treatment time. For this reason, the indication to remove displaced maxillary canines should not be taken from panoramic radiographs alone. PMID- 17124564 TI - Mini- and micro-screws for temporary skeletal anchorage in orthodontic therapy. AB - OBJECTIVE: Mini- and micro-screws are of increasing interest in orthodontic therapy in cases where maximum anchorage is requested. In this study we aimed to identify the criteria that influence the clinical success of micro-implants. MATERIAL AND METHODS: 239 micro-implants were inserted for skeletal anchorage in 85 patients undergoing orthodontic therapy. The survival rates of the first 133 implants were analyzed and those results were later applied in treating subsequent patients. RESULTS: In so doing, we were able to reduce the loss rate from 23% to 5%. Using thicker implants on the labial side of the lower jaw and the lingual side of the upper jaw was an important factor in reducing failure rates. We found the lower jaw's lingual side to be an inadequate region for inserting micro-implants. Thinner micro-implants were selected for the upper jaw's labial side. CONCLUSION: By following these guidelines, practitioners may find mini- and micro-screws to offer reliable orthodontic support. PMID- 17124565 TI - Maximum anchorage in orthodontics with the palatal implant: maximum anchorage in orthodontics with the palatal implant. AB - Maximum anchorage in the maxilla can only be achieved by using headgear worn full time or by using an implant. As full time wear requires considerable patient cooperation, we describe a technique for use of the Straumann Ortho Implant, exemplified by a case presentation. The cephalometric analysis at the end of therapy confirms that the upper molars did not slip forward during treatment (no anchorage loss). PMID- 17124566 TI - H2O2 but not O2- elevated by oxidized LDL enhances human aortic smooth muscle cell proliferation. AB - The oxidation of low density lipoprotein (LDL) as a key event in atherosclerosis suggests that antioxidant interventions may reduce the risk of atherosclerosis. However, the better strategies among antioxidant remedies for atherosclerosis remains difficult to be determined. Here, we show that oxidized LDL increases the steady-state level of intracellular hydrogen peroxide through stimulating the protein expressions of Nox1 and Cu/Zn superoxide dismutase (SOD) in human aortic smooth muscle cells (SMCs). The intracellular content of hydrogen peroxide rather than superoxide is a key modulator for vascular SMC (VSMC) proliferation, implying that without co-expression of catalase, increased Cu/Zn-SOD activity alone may not be beneficial to reduce the growth of VSMC in an atherosclerotic plaque. PMID- 17124567 TI - Stable carbon and nitrogen isotope ratio profiling of sperm whale teeth reveals ontogenetic movements and trophic ecology. AB - Teeth from male sperm whales (Physeter macrocephalus) stranded in the North eastern Atlantic were used to determine whether chronological profiles of stable isotope ratios of C (delta(13)C) and N (delta(15)N) across dentine growth layers could be used to detect known ontogenetic benchmarks in movements and trophic ecology. Profiles showed a general decrease in delta(13)C (median = 1.91 per thousand) and an increase in delta(15)N (median = 2.42 per thousand) with age. A marked decline in delta(13)C occurred for all 11 teeth around 9-10 years and again for six individuals around 20 years. After the early twenties the delta(13)C continued to decline with age for all teeth. These results are consistent with males segregating from natal groups in low latitudes with the onset of puberty between 4 and 15 years and gradually dispersing pole-ward into (13)C-depleted temperate waters. Penetration into further depleted, productive high latitudes after the age of 20 might facilitate the spurt of accelerated growth rate observed around this age. Breeding migrations back to lower latitudes were not reflected in the delta(13)C profiles possibly due to being short compared to the time spent feeding in high latitudes. The timings of marked isotopic change in the delta(15)N profiles reflect those of the delta(13)C profiles, suggesting a link between dietary changes and movements. The observed increase in delta(15)N with age is likely to be caused by a trophic level increase as males grow in size, probably feeding on larger prey. An additional explanation could be that, in the higher latitudes of the North Atlantic, the main prey source is the high trophic level squid Gonatus fabricii. Also, the lower latitudes from where males disperse are depleted in basal (15)N. Profiles of delta(13)C and delta(15)N in sperm whale teeth gathered from different regions, sexes, and periods in time, could provide a unique way to understand the ecology of this species across different oceans. PMID- 17124568 TI - Genetic variation in leaf pigment, optical and photosynthetic function among diverse phenotypes of Metrosideros polymorpha grown in a common garden. AB - Coordinated variation has been reported for leaf structure, composition and function, across and within species, and theoretically should occur across populations of a species that span an extensive environmental range. We focused on Hawaiian keystone tree species Metrosideros polymorpha, specifically, 13-year old trees grown (2-4 m tall) in a common garden (approximately 1 ha field with 2 3 m between trees) from seeds collected from 14 populations along an altitude soil age gradient. We determined the genetic component of relationships among specific leaf area (SLA), the concentrations of nitrogen (N) and pigments (chlorophylls, carotenoids, and anthocyanins), and photosynthetic light-use efficiency. These traits showed strong ecotypic variation; SLA declined 35% with increasing source elevation, and area-based concentrations of N, Chl a + b and Car increased by 50, 109 and 96%, respectively. Concentrations expressed on a mass basis were not well related to source elevation. Pigment ratios expressed covariation that suggested an increased capacity for light harvesting at higher source elevation; Chl/N and Car/Chl increased with source elevation, whereas Chl a/b declined; Chl a/b was higher for populations on younger soil, suggesting optimization for low N supply. Parallel trends were found for the photosynthetic reactions; light-saturated quantum yield of photosystem II (Phi (PSII)) and electron transport rate (ETR) increased with source elevation. Correlations of the concentrations of photosynthetic pigments, pigment ratios, and photosynthetic function across the ecotypes indicated a stoichiometric coordination of the components of the light-harvesting antennae and reaction centers. The constellation of coordinated morphological, biochemical and physiological properties was expressed in the leaf reflectance and transmittance properties in the visible and near-infrared wavelength region (400-950 nm), providing an integrated metric of leaf status among and between plant phenotypes. PMID- 17124569 TI - Isotopic enrichment in a phloem-feeding insect: influences of nutrient and water availability. AB - The isotopic enrichment between an animal and its diet can vary among and within living systems, but the sources of variation are not yet fully understood. Some studies have found that diet quality or an animal's nutritional status can influence the degree of trophic enrichment, while others have dismissed nutrition as a contributing factor. We evaluated the effects of nutrient and water availability on carbon and nitrogen isotopic enrichment in a specialized plant herbivore system. Aphids are largely sedentary and rely exclusively on nitrogen poor phloem sap of their host for nutrition. We grew potato aphids [Macrosiphum euphorbiae (Aphididae)] on an accepted host, pumpkin [Cucurbita pepo L. (Cucurbitaceae)], in a glasshouse environment. Twelve pumpkin plants growing under high- and low-watering regimes were inoculated at 4 weeks of age with aphids. During the course of the experiment we collected leaves, phloem sap, aphids and honeydew (i.e., aphid exudates). We found no trophic enrichment between aphids and their phloem sap diet, but significant carbon enrichment of honeydew relative to aphids (2.5 per thousand) and phloem sap (2.1 per thousand). Honeydew was also enriched in nitrogen compared to the phloem sap (1.2 per thousand). Watering treatment had a substantial impact on trophic enrichment. Correlations among tissues, an indication of uniform trophic enrichment among samples, were significant only for the carbon isotopic composition, and then only for plants and aphids grown in the low-water treatment. Diet quality also influenced the degree of isotopic enrichment; trophic enrichment for both carbon and nitrogen isotopic composition increased as diet quality (C/N) declined. We conclude that the degree of trophic enrichment is variable due, in part, to diet quality, but that the scale of variation is small. PMID- 17124570 TI - Experimental increase of flying costs in a pelagic seabird: effects on foraging strategies, nutritional state and chick condition. AB - A central point in life history theory is that parental investment in current reproduction should be balanced by the costs in terms of residual reproductive value. Long-lived seabirds are considered fixed investors, that is, parents fix a specific level of investment in their current reproduction independent to the breeding requirements. We tested this hypothesis analysing the consequences of an experimental increase in flying costs on the foraging ecology, body condition and chick condition in Cory's shearwaters Calonectris diomedea. We treated 28 pairs by reducing the wing surface in one partner and compared them with 14 control pairs. We monitored mass changes and incubation shifts and tracked 19 foraging trips per group using geolocators. Furthermore, we took blood samples at laying, hatching and chick-rearing to analyse the nutritional condition, haematology, muscle damage and stable isotopes. Eighty-day-old chicks were measured, blood sampled and challenged with PHA immune assay. In addition, we analysed the effects of handicap on the adults at the subsequent breeding season. During incubation, handicapped birds showed a greater foraging effort than control birds, as indicated by greater foraging distances and longer periods of foraging, covering larger areas. Eighty-day-old chicks reared by treated pairs were smaller and lighter and showed a lower immunity than those reared by control pairs. However, oxygen demands, nutritional condition and stable isotopes did not differ between control and handicapped birds. Although handicapped birds had to increase their foraging effort, they maintained physical condition by reducing parental investment and transferred the experimentally increased costs to their partners and the chick. This result supports the fixed investment hypothesis and is consistent with life history theory. PMID- 17124571 TI - [Coping with pain]. AB - Chronic pain is to be understood as a complex syndrome influenced by its intensity, character, localisation and time aspects as well as the subjective feeling of impairment and the accompanying emotions and cognitions. Based on the biopsychosocial concept of pain, therapies have to be multimodal to take into account the various dimensions of pain. One element of multimodal pain therapy is pain coping skills training, aimed at giving pain patients increasingly greater control over their pain by following the didactic guideline of small steps at a time. In addition to giving them disease-specific information, patients are supposed to learn to recognize dysfunctional components of their pain disorder and to alter them. Improved health and increased joy of living as well as a decrease of stressor-related activation (pain being the stressor) are the objectives of the training. Coping with pain as a programme aims at helping patients to help themselves. Pain coping skills training is indicated for patients already suffering from chronic pain or running a risk of pain chronicity. PMID- 17124572 TI - [Perioperative pain management in orthopaedics]. AB - Organized orthopaedic pain management is a major part of successful patient treatment. Therefore pain management should start before surgery. Patients need to be informed about the operation and the subsequent procedures. Clinical pain management is based on continuous pain documentation with pain as the fifth vital sign. Surgery should be minimally invasive bewaring a peripheral modulated nociceptive sensitization. In order to prevent chronic pain preemptive analgesia should be employed followed by an individually tailored regimen of post-operative analgesia. In consideration of the documented pain levels post-operative pain therapy consisting of a standing medication and a rescue medication should by adjusted daily. Due to the fact that the highest pain levels after surgery were reported within the first 48 h pain medication should be reduced in the ensuing days, again taking the documented pain levels into account. Supportive treatment approaches such as cryotherapy or transcutaneous electrical nerve stimulation (TENS) are useful in the post-operative period. Physiotherapy after surgery should be extended stepwise regarding the operative device and it is of particular importance to respect pain intensities. The post-hospital regimen for a continuous pain medication should be given to the orthopaedic specialist. PMID- 17124573 TI - Cavities generated by self-organised monolayers as sensitive coatings for surface acoustic wave resonators. AB - Silanisation of quartz substrate surfaces with a mixture of two chlorosilanes, namely trimethylchlorosilane and 7-octenyldimethylchlorosilane, leads to sensitive coatings for volatile organic compounds (VOC) on surface acoustic wave (SAW) devices. In this way we created monolayers of molecular cavities engulfing the analytes according to host-guest chemistry directly on the device surfaces, and also confirmed the occurrence of such cavities by molecular modelling. We monitored the binding process of the silanes by using Fourier transform infrared (FTIR) spectrometry and atomic force microscopy (AFM). In order to increase the stiffness of the cavities, we crosslinked the terminal double bonds of the long spacers by heating the surface in the presence of a radical initiator. Compared to SAW delay lines silanised with trimethylchlorosilane, devices modified with the binary silane mixture lead to substantially higher frequency shifts when exposed to solvent vapour streams. Nearly instantaneous responses can be observed, which e.g. allows xylene detection down to a few ppm. PMID- 17124574 TI - A theoretical framework for evaluating analytical digestion methods for poorly soluble particulate beryllium. AB - Complete digestion of all chemical forms and sizes of particulate analytes in environmental samples is usually necessary to obtain accurate results with atomic spectroscopy. In the current study, we investigate the physicochemical properties of beryllium particles likely to be encountered in samples collected from different occupational environments and present a hypothesis that a dissolution theory can be used as a conceptual framework to guide development of strategies for digestion procedures. For monodisperse single-chemical constituent primary particles, such as those encountered when handling some types of beryllium oxide (BeO) powder, theory predicts that a digestion procedure is sufficient when it completely dissolves all primary particles, independent of cluster size. For polydisperse single-chemical constituent particles, such as those encountered during the handling of some types of beryllium metal powder, theory predicts that a digestion procedure is sufficient only when it completely dissolves the largest particle in the sample. For samples with unknown or multi-chemical constituent particles and with particles having undefined sizes, e.g., fume emissions from a copper-beryllium alloy furnace operation or dust from a beryl ore crushing operation, a surface area-limited and single-constituent-dependent dissolution theory may not predict complete dissolution, thereby requiring non-routine robust treatment procedures with post-digestion filtration, followed by examination of residual particulate material. Additionally, for beryllium, and likely other poorly soluble materials, particulate reference materials of various chemical forms and size distributions are needed to better evaluate and harmonize analytical digestion procedures. Figure Generation of aerosol particles during machining of beryllium oxide. PMID- 17124575 TI - Laser-induced breakdown spectroscopy (LIBS) in archaeological science- applications and prospects. AB - Laser-induced breakdown spectroscopy (LIBS) has emerged in the past ten years as a promising technique for analysis and characterization of the composition of a broad variety of objects of cultural heritage including painted artworks, icons, polychromes, pottery, sculpture, and metal, glass, and stone artifacts. This article describes in brief the basic principles and technological aspects of LIBS, and reviews several test cases that demonstrate the applicability and prospects of LIBS in the field of archaeological science. PMID- 17124576 TI - Contraindication labelling changes in the United States and Germany. AB - OBJECTIVE: Communication of a drug's benefits and risks through drug labelling is an important tool in the risk management of prescription drugs. Contraindication labelling provides the strongest safety-based guidance for the health care practitioner. In this study, the labelling of new contraindications between the United States (US) and Germany was compared. METHODS: We assessed whether contraindications that were newly labelled in the US between January 2003 and May 2005 had been incorporated into the German Summary of Product Characteristics (SPC) by October 2005 (end of the study period). We classified the results of our comparison as no difference, a slight difference (more detailed contraindication in one country or the other) or as a relevant difference (new US contraindication not contraindicated in the German SPC). We assessed the time difference between the date of the labelling change in the US and October 2005. RESULTS: During the study period, 66 new contraindications were evaluable for analysis. We found no difference in labelling between the US and Germany with regard to 30 contraindications (45.5%) and slight differences with regard to 14 contraindications (21.2%). In 22 instances (33.3%), the new contraindication in the US had not been incorporated into the German SPC. The median delay between the date of labelling in the US and October 2005 was 14 months and ranged between 5 and 31 months. In 77.3% of the relevant differences in contraindication labelling, the pharmaceutical manufacturer was the same in the US and Germany. CONCLUSION: Our study shows a remarkable discrepancy in the labelling of new contraindications between the US and Germany. The underlying reasons for this discrepancy were discussed, but need to be explored further. PMID- 17124577 TI - The awareness among paediatricians of off-label prescribing in children: a survey of Italian hospitals. AB - OBJECTIVE: To investigate paediatricians' perception and awareness of off-label (OL) and unlicensed drug usage in clinical practice. METHODS: We conducted a questionnaire-based, observational study involving 28 departments/specialty units of four tertiary paediatrics Italian hospitals. The questionnaire was sent to a designated paediatrician in each department/specialty unit with the request that they summarise their awareness of OL prescriptions after a group discussion with all paediatricians working in the same department/unit. RESULTS: A total of 95 drugs were identified. The most common perceived reasons detected for OL use were age and formulation. Corticosteroids, ACE-inhibitors, beta-blockers and calcium antagonists were most frequently indicated as OL for formulation, while ACE inhibitors, new generation anti-epilepsy drugs, immunosuppressants, ketorolac and propofol as OL for age. Adrenaline, midazolam and ACE-inhibitors were indicated as unlicensed for route of administration. We found a high concordance between the OL drugs identified by physicians and those reported in the "Italian Summary Product Characteristics". CONCLUSIONS: Italian paediatricians perceived different drugs as unlicensed/OL from those identified by existing prescriptions studies. These perceived drugs may represent a priority in tackling the problem of OL usage in the hospital practice. The review of the Italian Summary Product Characteristics of some of the drugs identified, together with the monitoring of permanent clinical practice and with new clinical research, may be a step forward to reduce OL use in children. PMID- 17124578 TI - Terbinafine increases the plasma concentration of paroxetine after a single oral administration of paroxetine in healthy subjects. AB - OBJECTIVE: Paroxetine is believed to be a substrate of CYP2D6. However, no information was available indicating drug interaction between paroxetine and inhibitors of CYP2D6. The aim of this study was to examine the effects of terbinafine, a potent inhibitor of CYP2D6, on pharmacokinetics of paroxetine. METHODS: Two 6-day courses of either a daily 150-mg of terbinafine or a placebo, with at least a 4-week washout period, were conducted. Twelve volunteers took a single oral 20-mg dose of paroxetine on day 6 of both courses. Plasma concentrations of paroxetine were monitored up to 48 h after dosing. RESULTS: Compared with the placebo, terbinafine treatment significantly increased the peak plasma concentration (C(max)) of paroxetine, by 1.9-fold (6.4 +/- 2.4 versus 12.1 +/- 2.9 ng/ml, p < 0.001), and the area under the plasma concentration-time curve from zero to 48 h [AUC (0-48)] of paroxetine by 2.5-fold (127 +/- 67 vs 318 +/- 102 ng/ml, p < 0.001). Elimination half-life differed significantly (15.3 +/- 2.4 vs 22.7 +/- 8.8 h, p < 0.05), although the magnitude of alteration (1.4-fold) was smaller than C(max )or AUC. CONCLUSION: The present study demonstrated that the metabolism of paroxetine after a single oral dose was inhibited by terbinafine, suggesting that inhibition of CYP2D6 activity may lead to a change in the pharmacokinetics of paroxetine. However, further study is required to confirm this phenomenon at steady state. PMID- 17124579 TI - Optimization of culture conditions and scale-up to pilot and plant scales for coenzyme Q10 production by Agrobacterium tumefaciens. AB - This report describes the optimization of culture conditions for coenzyme Q(10) (CoQ(10)) production by Agrobacterium tumefaciens KCCM 10413, an identified high CoQ(10)-producing strain (Kim et al., Korean patent. 10-0458818, 2002b). Among the conditions tested, the pH and the dissolved oxygen (DO) levels were the key factors affecting CoQ(10) production. When the pH and DO levels were controlled at 7.0 and 0-10%, respectively, a dry cell weight (DCW) of 48.4 g l(-1) and a CoQ(10) production of 320 mg l(-1) were obtained after 96 h of batch culture, corresponding to a specific CoQ(10) content of 6.61 mg g-DCW(-1). In a fed-batch culture of sucrose, the DCW, specific CoQ(10) content, and CoQ(10) production increased to 53.6 g l(-1), 8.54 mg g-DCW(-1), and 458 mg l(-1), respectively. CoQ(10) production was scaled up from a laboratory scale (5-l fermentor) to a pilot scale (300 l) and a plant scale (5,000 l) using the impeller tip velocity (V (tip)) as a scale-up parameter. CoQ(10) production at the laboratory scale was similar to those at the pilot and plant scales. This is the first report of pilot and plant-scale productions of CoQ(10) in A. tumefaciens. PMID- 17124580 TI - Enhanced vanillin production from ferulic acid using adsorbent resin. AB - High vanillin productivity was achieved in the batch biotransformation of ferulic acid by Streptomyces sp. strain V-1. Due to the toxicity of vanillin and the product inhibition, fed-batch biotransformation with high concentration of ferulic acid was unsuccessful. To solve this problem and improve the vanillin yield, a biotransformation strategy using adsorbent resin was investigated. Several macroporous adsorbent resins were chosen to adsorb vanillin in situ during the bioconversion. Resin DM11 was found to be the best, which adsorbed the most vanillin and the least ferulic acid. When 8% resin DM11 (wet w/v) was added to the biotransformation system, 45 g l(-1) ferulic acid could be added continually and 19.2 g l(-1) vanillin was obtained within 55 h, which was the highest vanillin yield by bioconversion until now. This yield was remarkable for exceeding the crystallization concentration of vanillin and therefore had far reaching consequence in its downstream processing. PMID- 17124581 TI - Taxanes: perspectives for biotechnological production. AB - Taxol is a valuable plant-derived drug showing activity against various cancer types. Worldwide efforts had been made to overcome the supply problem, because the supply by isolation from the bark of the slow-growing yew trees is limited. Plant cell cultures as well as chemical and biotechnological semisynthesis are processes, which are intensively investigated for the production of taxanes paclitaxel (Taxol) and docetaxel (Taxotere) in the last few years. This article provides a comparison of the current research on taxane biosynthesis and production in yew cell cultures. PMID- 17124582 TI - Translation elongation factor 1-alpha gene from Pichia pastoris: molecular cloning, sequence, and use of its promoter. AB - The gene encoding translation elongation factor 1-alpha from the yeast Pichia pastoris was cloned. The gene revealed an open reading frame of 1,380 bp with the potential to encode a polypeptide of 459 amino acids with a calculated mass of 50.1 kDa. The potential of the promoter (P (TEF1)) in P. pastoris was investigated with comparison to the glyceraldehyde-3-phosphate dehydrogenase promoter (P (GAP)) by using a bacterial lipase gene as a reporter gene. P (TEF1) demonstrated a tighter growth-associated expression mode, improved functioning in the presence of high glucose concentrations, and promoter activities that yielded recombinant protein at levels similar to or in one case greater than P (GAP). The sequence of the gene was deposited in GenBank under accession no. EF014948. PMID- 17124583 TI - Proteomic profile changes in membranes of ethanol-tolerant Clostridium thermocellum. AB - Clostridium thermocellum, a cellulolytic, thermophilic anaerobe, has potential for commercial exploitation in converting fibrous biomass to ethanol. However, ethanol concentrations above 1% (w/v) are inhibitory to growth and fermentation, and this limits industrial application of the organism. Recent work with ethanol adapted strains suggested that protein changes occurred during ethanol adaptation, particularly in the membrane proteome. A two-stage Bicine-doubled sodium dodecyl sulfate-polyacrylamide gel electrophoresis protocol was designed to separate membrane proteins and circumvent problems associated with membrane protein analysis using traditional gel-based proteomics approaches. Wild-type and ethanol-adapted C. thermocellum membranes displayed similar spot diversity and approximately 60% of proteins identified from purified membrane fractions were observed to be differentially expressed in the two strains. A majority (73%) of differentially expressed proteins were down-regulated in the ethanol-adapted strain. Based on putative identifications, a significant proportion of these down regulated proteins were involved with carbohydrate transport and metabolism. Approximately one-third of the up-regulated proteins in the ethanol-adapted species were associated with chemotaxis and signal transduction. Overall, the results suggested that membrane-associated proteins in the ethanol-adapted strain are either being synthesized in lower quantities or not properly incorporated into the cell membrane. PMID- 17124584 TI - Comprehensive epitope mapping of the Epstein-Barr virus latent membrane protein-2 in normal, non tumor-bearing individuals. AB - Latent membrane protein (LMP)-2 is one of the Epstein-Barr virus (EBV)-encoded proteins consistently expressed by nasopharyngeal carcinoma (NPC). EBV transformed lymphoblastoid cell lines (LCL) have been used in patients with NPC to induce LMP-2-recognizing T cell lines which have been in turn utilized for protein-wide mapping of T cell epitopes. However, comprehensive mapping of naturally recognized LMP-2 epitopes in non tumor-bearing individuals has not been reported. Here, we applied a low sensitivity epitope-defining technique for the identification of LMP-2 CTL responses detectable ex vivo in EBV-experienced individuals. This screening tool has been previously validated by analyzing memory CTL responses to Flu, cytomegalovirus (CMV), and the melanoma associated antigen gp100/Mel17. Peripheral blood monocytes (PBMC) from ten Caucasian and ten Chinese individuals were stimulated ex vivo with pools of nonamer (9-mer) peptides overlapping in a stepwise fashion each single amino acid of the LMP-2 sequence. No obvious differences were observed between the immune response of the two ethnic groups save for those related to the divergence in the ethnic prevalence of HLA haplotypes. Several novel and known LMP-2 epitopes were identified. Reactivity toward at least one LMP-2 epitope was detected in 18 of the 20 donors but no prevalent human leukocyte antigen (HLA)/epitope combination was observed confirming that LMP-2 reactivity in the context of common HLA alleles is more pleiotropic than that of FLU and CMV. We believe that the usefulness of these epitopes occurring naturally in non-cancer bearing patients as reagents for the immunization of patients with early or advanced stage NPC deserves further evaluation. PMID- 17124585 TI - Survivin expression in "low-risk" and "high-risk" myelodysplastic syndromes. AB - Apoptosis has a crucial role in myelodysplastic syndromes (MDS), being responsible of the ineffective hematopoiesis characteristic of the disease. Apoptosis rate is elevated in "early phase" MDS, whereas it diminishes during disease progression to acute leukemia, consensually to the acquisition of independent growth features. Survivin is a member of the inhibitor of the apoptosis (IAP) family, with the bifunctional role of suppressing apoptosis while facilitating cell cycle progression. We investigated Survivin mRNA levels by real time quantitative reverse transcriptase PCR analysis and Survivin protein expression by immunohistochemistry in 49 bone marrow (BM) aspirates and in 17 BM biopsies (BMB) from MDS patients. Survivin mRNA levels were higher in MDS than in control group (1.68 +/- 1.46 vs 0.25 +/- 0.22; p < 0.0001). MDS patients with low or INT1 International Scoring System for Evaluating Prognosis (IPSS) displayed higher levels of Survivin mRNA in comparison to INT2 or high IPSS (1.91 +/- 1.51 vs 0.88 +/- 0.95; p = 0.0058). Survivin protein immunoreactivity was evaluated as Survivin index S ((i)) and calculated according to the formula: S ((i)) = % of Survivin positive cells x BMB cellularity / 100. Survivin index was higher in the MDS group than in normal BM (p = 0.05). Moreover, in eight cases in which BM aspirates and trephine biopsy were available, we found a significant association between the level of Survivin mRNA and protein expression (p = 0.011). In conclusion, this study demonstrates increased levels of Survivin in MDS compared to normal controls. Moreover, higher levels of transcripts are related to "low risk" MDS. Our results suggest an active role of Survivin in normal and in myelodysplastic hematopoiesis. PMID- 17124586 TI - Serum anti-Fcgamma receptor autoantibodies in patients with alopecia areata. AB - Although anti-Fcgamma receptor antibodies (Abs) are detected in various autoimmune diseases, there have been no studies about the anti-Fcgamma receptor Abs in alopecia areata (AA). To detect the anti-Fcgamma receptor Abs in patients with AA and their clinical correlations, Serum samples from 72 patients with AA and 23 normal controls were examined by enzyme-linked immunosorbent assay assessing anti-Fcgamma receptor Ab levels. Anti-Fcgamma receptor I Abs were significantly frequently detected in patients with AA compared with normal controls. Furthermore, the detection of anti-Fcgamma receptor I Abs significantly inversely correlated with the disease duration. These results suggest that anti Fcgamma receptor I Ab and Fcgamma receptor I play an important role in the regulation of AA, are useful for a marker of the disease prognosis and are worth intense research for the reasonable and specific therapy of AA. PMID- 17124587 TI - Germline mutation in the STK11 gene in a girl with an ovarian Sertoli cell tumour. AB - INTRODUCTION: An ovarian Sertoli cell tumour was detected in a 4-year-old girl with gonadotrophin-independent precocious puberty. Such gonadal tumours can be associated with Peutz-Jeghers syndrome, caused by mutations in the STK11 gene. We have therefore sequenced the STK11 gene. RESULTS: Mutation analysis revealed a nonsense mutation in exon 1 (c.130A>T;p.Lys44X) of the SKT11 gene, which resulted in a truncated, inactive protein. The mutation was heterozygous in patient's lymphocytes and almost homozygous in the tumour, indicating loss of heterozygosity. CONCLUSION: This is the first report of a STK11 germline mutation in a girl with an ovarian Sertoli cell tumour. It remains to be shown whether this particular mutation predisposes the patient to the development of ovarian tumours. PMID- 17124588 TI - Hemothorax as presentation of late vitamin-K-deficient bleeding in a 1-month-old infant with homozygous alpha-1-antitrypsin deficiency. PMID- 17124589 TI - Potato crisps without pancreatic extracts supplements: a potential cause of the distal intestinal obstruction in cystic fibrosis. AB - Three cases of the distal intestinal obstruction syndrome (DIOS) are reported to be caused by a consumption of potato crisps without appropriate pancreatic enzyme supplements. PMID- 17124590 TI - Leaf wetness duration measurement: comparison of cylindrical and flat plate sensors under different field conditions. AB - In general, leaf wetness duration (LWD) is a key parameter influencing plant disease epidemiology, since it provides the free water required by pathogens to infect foliar tissue. LWD is used as an input in many disease warning systems, which help growers to decide the best time to spray their crops against diseases. Since there is no observation standard either for sensor or exposure, LWD measurement is often problematic. To assess the performance of electronic sensors, LWD measurements obtained with painted cylindrical and flat plate sensors were compared under different field conditions in Elora, Ontario, Canada, and in Piracicaba, Sao Paulo, Brazil. The sensors were tested in four different crop environments--mowed turfgrass, maize, soybean, and tomatoes--during the summer of 2003 and 2004 in Elora and during the winter of 2005 in Piracicaba. Flat plate sensors were deployed facing north and at 45 degrees to horizontal, and cylindrical sensors were deployed horizontally. At the turfgrass site, both sensors were installed 30 cm above the ground, while at the crop fields, the sensors were installed at the top and inside the canopy (except for maize, with a sensor only at the top). Considering the flat plate sensor as a reference (Sentelhas et al. Operational exposure of leaf wetness sensors. Agric For Meteorol 126:59-72, 2004a), the results in the more humid climate at Elora showed that the cylindrical sensor overestimated LWD by 1.1-4.2 h, depending on the crop and canopy position. The main cause of the overestimation was the accumulation of big water drops along the bottom of the cylindrical sensors, which required much more energy and, consequently, time to evaporate. The overall difference between sensors when evaporating wetness formed during the night was around 1.6 h. Cylindrical sensors also detected wetness earlier than did flat plates--around 0.6 h. Agreement between plate and cylinder sensors was much better in the drier climate at Piracicaba. These results allow us to caution that cylindrical sensors may overestimate wetness for operational LWD measurements in humid climates and that the effect of other protocols for angling or positioning this sensor should be investigated for different crops. PMID- 17124591 TI - The larval parasitoid Microplitis croceipes oviposits in conspecific adults. AB - Microplitis croceipes (Hymenoptera: Braconidae) is a larval parasitoid of Helicoverpa/Heliothis spp. In the course of mass rearing of M. croceipes, we found that females oviposited in the conspecific adults in rearing cages. When 20 pairs of inexperienced females and males or of experienced females and males were reared in a cage, the males lived for 14-15 days and the females for 18-20 days on average. At their death, 37-42% of the males and 50-57% of the females contained conspecific eggs or first instar larvae in their abdominal cavity. When two of inexperienced females met on a host-infested leaf of soybean, they attempted to sting each other. Of the attacked females, 30% contained a conspecific egg laid in their abdomen. In abdominal cavity of the adults parasitized by a conspecific female, the majority of the parasitoid eggs laid disappeared within 1 day after oviposition. Only 10-30% of the parasitoid eggs laid in conspecific adults hatched 3-4 days after oviposition, but those larvae never molted to second instar. When the adults were stung by one or two conspecific females, their subsequent longevity was significantly shorter than that for the control adults. Oviposition in conspecific adults may be prevalent in other parasitic wasps that quickly oviposit without intensive host examination, and have cuticle and size of abdomen to be stung by conspeicifcs. PMID- 17124592 TI - Proteomic analysis of phytopathogenic fungus Botrytis cinerea as a potential tool for identifying pathogenicity factors, therapeutic targets and for basic research. AB - Botrytis cinerea is a phytopathogenic fungus causing disease in a substantial number of economically important crops. In an attempt to identify putative fungal virulence factors, the two-dimensional gel electrophoresis (2-DE) protein profile from two B. cinerea strains differing in virulence and toxin production were compared. Protein extracts from fungal mycelium obtained by tissue homogenization were analyzed. The mycelial 2-DE protein profile revealed the existence of qualitative and quantitative differences between the analyzed strains. The lack of genomic data from B. cinerea required the use of peptide fragmentation data from MALDI-TOF/TOF and ESI ion trap for protein identification, resulting in the identification of 27 protein spots. A significant number of spots were identified as malate dehydrogenase (MDH) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The different expression patterns revealed by some of the identified proteins could be ascribed to differences in virulence between strains. Our results indicate that proteomic analysis are becoming an important tool to be used as a starting point for identifying new pathogenicity factors, therapeutic targets and for basic research on this plant pathogen in the postgenomic era. PMID- 17124593 TI - Dexamethasone as a probe for CYP3A4 metabolism: evidence of gender effect. AB - BACKGROUND: A study was conducted to evaluate prospectively the correlation between docetaxel clearance and pharmacokinetics of dexamethasone previously obtained in 21 patients. PATIENTS AND METHODS: Dexamethasone pharmacokinetics were performed in 17 patients 24 h before docetaxel treatment as monochemotherapy. Dexamethasone and docetaxel plasma concentrations were determined by HPLC methods. Determination of docetaxel unbound fraction in plasma was performed using microequilibrium dialysis. RESULTS: Significant correlation was observed between observed plasma docetaxel clearances (CL(docetaxel)) and values predicted from dexamethasone plasma clearance (CL(dexa)), unbound plasma docetaxel fraction estimated from serum alpha1-acid glycoprotein level (fu(alpha1 AAG)), and hepatic metastasis status. However, after splitting of the prospective data set according to gender, no correlation was observed for males (R(2) = 0.08, NS, n = 10), then strong correlation was observed for females (R(2) = 0.78, P < 0.01, n = 7). Multivariate analysis was performed from data obtained in the women included in the first study and those of this prospective study (n = 18). Docetaxel CL was significantly correlated with CL(dexa) (P = 0.001) and fu(alpha1 AAG) (P = 0.01) according to the relationship (with +/-95% confidence intervals): CL(docetaxel) (l/h) = 1.92 (+/-0.94) x CL(dexa) (l/h) + 2.68 (+/-1.95) x fu(alpha1-AAG) (%) (R(2) = 0.68). CONCLUSION: Dexamethasone may be used to predict docetaxel clearances in females, but not in males. PMID- 17124594 TI - The histone deacetylase inhibitor PXD101 synergises with 5-fluorouracil to inhibit colon cancer cell growth in vitro and in vivo. AB - PURPOSE: Histone deacetylase inhibitors (HDACi) inhibit the growth of cancer cells, and combinations of HDACi with established chemotherapeutics can lead to synergistic effects. We have investigated effects of PXD101 (HDACi in phase II clinical trials) in combination with 5-fluorouracil, on tumour cell proliferation and apoptosis both in vitro and in vivo. EXPERIMENTAL DESIGN: HCT116 cells were studied using proliferation and clonogenic assays. Synergistic inhibition of proliferation and clonogenicity was determined by incubation with PXD101 and 5 fluorouracil, and analysis using CalcuSyn software. The effect of combining PXD101 and 5-fluorouracil on apoptosis was examined in vitro using PARP-cleavage and TUNEL. Finally, the effectiveness of combining PXD101 and 5-fluorouracil in vivo was tested using both HT-29 and HCT116 xenograft models. RESULTS: Synergistic inhibition of proliferation and clonogenicity was obtained when HCT116 cells were incubated with PXD101 and 5-fluorouracil. 5-fluorouracil combined with PXD101 also increased DNA fragmentation and PARP cleavage in HCT116 cells. Incubation with PXD101 down regulated thymidylate synthase expression in HCT116 cells. In vivo studies, using mouse HT29 and HCT116 xenograft models, showed improved reductions in tumour volume compared to single compound, when PXD101 and 5-fluorouracil were combined. CONCLUSIONS: PXD101 and 5-fluorouracil synergistically combine in their anti-tumour effects against colon cancer cells in vitro and show enhanced activity when combined in vivo. Based on the results presented herein, a rationale for the use of PXD101 and 5-fluorouracil in combination in the clinic has been demonstrated. PMID- 17124595 TI - Multicentre phase II study using increasing doses of irinotecan combined with a simplified LV5FU2 regimen in metastatic colorectal cancer. AB - PURPOSE: Irinotecan at 180 mg/m2 combined with an infusional 5 fluorouracil/leucovorin (5-FU/LV) regimen (FOLFIRI) is a standard first line therapy for metastatic colorectal cancer (mCRC). This phase II study aimed to assess whether increasing the irinotecan dose in the first line FOLFIRI regimen would benefit mCRC patients. PATIENTS AND METHODS: Patients received FOLFIRI every 2 weeks for up to six cycles, comprising a 5-FU/LV regimen combined with irinotecan at 180 mg/m2 (cycle 1), increasing to 220 mg/m2 (cycle 2) and 260 mg/m2 (cycle 3 and subsequent cycles) dependent on toxicity. Efficacy and safety were determined in the intention to treat (ITT) population and in patients able to receive irinotecan at 260 mg/m2 for at least four cycles [high-dose (HD) population]. RESULTS: Fifty-four eligible patients were included. Among them, 44 (81.5%) formed the HD population. The ITT objective response rate was 48% (90%CI: 36-60) with 25/26 of the responses in the HD population. The disease control rate was 76% (90%CI: 65-85) and median overall survival was 20.4 months (90%CI: 6.4 27.1). The main grade 3/4 toxicities (ITT/HD populations) were neutropenia (61%/59%), and diarrhoea (18%/11%), respectively. CONCLUSIONS: This study confirms the feasibility of increasing the standard dose of the irinotecan component of FOLFIRI to 260 mg/m2, for more than 80% of patients but does not support a clear advantage of this strategy on unselected mCRC patients. PMID- 17124596 TI - The efficacy and relationship between peak concentration and toxicity profile of fixed-dose-rate gemcitabine plus carboplatin in patients with advanced non-small cell lung cancer. AB - PURPOSE: To investigate the efficacy and relationship between plasma concentrations at the end of infusion (C(end of infusion)) and toxicity profile of fixed-dose-rate gemcitabine plus carboplatin in chemotherapy-naive patients with advanced non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Patients were given gemcitabine by 120 min infusion [at a fixed dose rate (FDR) of 10 mg/m(2)/min] on days 1 and 8 of a 21-day cycle, immediately followed by carboplatin AUC 5 by 4 h infusion on day 1. C (end of infusion) of gemcitabine was determined by ion-pair reversed-phase high-performance liquid chromatography (HPLC). RESULTS: By the close-out date, in our study population, the estimated median time to tumor progression (TTP) was 7 months (95% CI 4-10 months), median overall survival (OS) was 12 months (95% CI 11.2-12.8 months). The mean value of C (end of infusion) of 21 eligible patients was 16.48 +/- 8.07 micromol/l (range 27.43-2.87 micromol/l). The main hematological toxicities were transient grade 3 4 thrombocytopenia. The mean percentages of reduction of WBC, NEC, PLTC and Hb of 21 eligible patients were 38.3 +/- 38.1%, 31.3 +/- 73.6%, 31.8 +/- 53.5% and 12.0 +/- 12.2%, respectively. The analysis of the C(end of infusion) of gemcitabine and the percentage of reduction in WBC showed a significant correlation (r(2) = 0.4575; p < 0.05). A significant correlation (r(2) = 0.5671; p < 0.05) was also observed between the percentage of reduction of PLTC and C(end of infusion) of gemcitabine infusion. CONCLUSION: The clinical data in this trial supports the further evaluation the regimen in advanced NSCLC patients, due to its predictable kinetic behavior and less severe toxicity profile than expected. PMID- 17124597 TI - Fabry disease and hearing loss. Comment on: Barras FM, Maire R (2006) Progressive hearing loss in Fabry's disease: a case report. Eur Arch Otorhinolaryngol 263:688 691. PMID- 17124598 TI - Herpes zoster laryngitis: case report and serological profile. AB - Compared to herpes zoster oticus, varicella zoster virus (VZV) reactivations in immunocompetent patients are rare in laryngeal region. Usually, associated vocal cord paralyses are reported. Herein is a case report of a patient with laryngeal zoster without any associated motor disorders. An attempt is made to assign the distribution of mucosal eruptions to the appropriate neuroanatomical structures. A description of the serological course of VZV IgM and IgG are provided. Vesicles were found on the left sensory distribution areas of the superior laryngeal nerve. VZV IgM and IgG antibodies reached their peak 1 month after initial symptoms. Attentive follow-up and no antiviral therapy were advocated because of the absence of any immune deficiency or endoscopic suspicion of malignancy. In this case of VZV reactivation in the sensitive area of the superior laryngeal nerve, serological profiles of VZV IgM and IgG were profoundly modified up to the fourth month. PMID- 17124599 TI - A renal cell carcinoma with components of both chromophobe and papillary carcinoma. AB - We report a case of a morphologically unusual renal cell carcinoma with features of both chromophobe and papillary carcinoma. Immunohistochemical analysis of high molecular weight cytokeratins (HMWCK), cytokeratin 7 (CK7), cytokeratin 19 (CK19), c-Kit, and alpha-methylacyl-CoA racemase (AMACR) demonstrated differential profiles for the two components of the tumor, consistent with the respective patterns commonly observed for pure chromophobe and papillary renal cell carcinomas. Specifically, the chromophobe tumor cells expressed CK7 and c Kit weakly, while HMWCK, CK19, and AMACR were not detectable. In contrast, the papillary tumor cells expressed uniformly HMWCK, CK7, and c-Kit and focally CK19 and AMACR. Fluorescence in situ hybridization analysis of nuclei isolated from paraffin-embedded tumor tissue detected monosomy 1, disomy 7, and monosomy 17, a common and characteristic finding in chromophobe carcinomas, in a majority of, but not all tumor cells, whereas a population characterized by disomy 1, trisomy 7, and trisomy 17, a frequent finding in papillary carcinoma, was not identifiable. Electron microscopic analysis revealed numerous characteristic small cytoplasmic vesicles in the chromophobe areas, which were absent in the papillary component. This case illustrates the rare coexistence of two distinct and admixed histologic types of renal cell carcinoma. PMID- 17124600 TI - Differing distributions of CXCR3- and CCR4-positive cells among types of interstitial pneumonia associated with collagen vascular diseases. AB - Interstitial pneumonia (IP) is an important complication in collagen vascular diseases (CVDs). We examined the distribution of helper T cell subsets in lung biopsies of cases of IP associated with CVD (CVD-IP). The tissues from 27 CVD-IP patients with rheumatoid arthritis (RA), 8 with polymyositis or dermatomyositis (PM/DM), and 8 with systemic sclerosis (SSc) were compared with those from 10 patients with idiopathic pulmonary fibrosis (IPF) in our previous study. The expressions of CXCR3 and CCR4 (chemokine receptors associated in vitro with Th1 and Th2 cells, respectively) in the mononuclear infiltrate were analyzed immunohistochemically. The positive cells were semiquantified in fibrosing areas of the CVD-IP and IPF cases. The number of CXCR3-positive cells was significantly greater in RA-IP than in PM/DM-IP, SSc-IP, or IPF, whereas there were fewer CCR4 positive cells in RA-IP, PM/DM-IP, and SSc-IP than in IPF. The CXCR3-/CCR4 positive cells ratio was significantly higher in RA-IP and PM/DM-IP (but not in SSc-IP) than in IPF. These results support previous reports of the dominance of Th2 cells in some SSc-IP and IPF cases. However, Th1-type immune responses may predominate in RA-IP and PM/DM-IP. Our findings suggest that the pathogenesis of CVD-IPs differs with the helper T cell subset. PMID- 17124601 TI - Sporadic subcutaneous angiomyolipoma with expression of estrogen and progesterone receptors. PMID- 17124602 TI - A model of cerebrocerebello-spinomuscular interaction in the sagittal control of human walking. AB - A computationally developed model of human upright balance control (Jo and Massaquoi on Biol cybern 91:188-202, 2004) has been enhanced to describe biped walking in the sagittal plane. The model incorporates (a) non-linear muscle mechanics having activation level -dependent impedance, (b) scheduled cerebrocerebellar interaction for control of center of mass position and trunk pitch angle, (c) rectangular pulse-like feedforward commands from a brainstem/ spinal pattern generator, and (d) segmental reflex modulation of muscular synergies to refine inter-joint coordination. The model can stand when muscles around the ankle are coactivated. When trigger signals activate, the model transitions from standing still to walking at 1.5 m/s. Simulated natural walking displays none of seven pathological gait features. The model can simulate different walking speeds by tuning the amplitude and frequency in spinal pattern generator. The walking is stable against forward and backward pushes of up to 70 and 75 N, respectively, and with sudden changes in trunk mass of up to 18%. The sensitivity of the model to changes in neural parameters and the predicted behavioral results of simulated neural system lesions are examined. The deficit gait simulations may be useful to support the functional and anatomical correspondences of the model. The model demonstrates that basic human-like walking can be achieved by a hierarchical structure of stabilized-long loop feedback and synergy-mediated feedforward controls. In particular, internal models of body dynamics are not required. PMID- 17124603 TI - Protein Z and natural anticoagulants in children on peritoneal dialysis and hemodialysis. AB - Hemostatic alterations due to abnormalities in the coagulation and fibrinolytic system may occur in dialysis patients. Protein Z (PZ) is a vitamin K-dependent coagulation protein promoting assembly of thrombin with phospholipid vesicles. The aim of this study was to investigate PZ and natural anticoagulants in children on hemodialysis (HD) and peritoneal dialysis (PD). Protein Z, protein C (PC), protein S (PS), antithrombin III (AT III), and fibrinogen levels were studied in 24 PD, 13 HD patients and 23 controls. Plasma PZ levels in patients on HD were significantly higher than those on PD and control group (p = 0.04, p = 0.03). We observed elevated PC, PS and AT III activities in children on PD when compared to controls (p = 0.011, p = 0.003, p < 0.001). In HD patients, only PS activity was increased compared to controls (p = 0.016). PC and PS activities did not differ between PD and HD patients whereas AT III activity was higher in PD patients compared to HD patients (p < 0.001). Normal/high levels of PC, PS and AT III suggest that children on PD or HD treatment do not seem to have an increased risk of thrombogenesis due to reduction of these proteins. Increased PZ levels, however, might contribute to the hemostatic alterations in children on HD treatment along with other well known abnormalities. PMID- 17124604 TI - Prognosis of autosomal dominant polycystic kidney disease diagnosed in utero or at birth. AB - The use of prenatal ultrasonography has resulted in increased numbers of fetuses being diagnosed with autosomal dominant polycystic kidney disease (ADPKD), but the long-term prognosis is still not well-known. Between 1981 and 2006 we followed 26 consecutive children with enlarged hyperechoic kidneys detected between the 12th week of pregnancy and the first day of life (Day 1) as well as one affected parent. Three other fetuses were excluded following the termination of the pregnancy. The mother was the transmitting parent in 16 of the 26 children (ns, p=0.1). Clinical features that presented during follow-up were oligoamnios (5/26), neonatal pneumothorax (3/26), pyelonephritis (5/26), gross hematuria (2/26), hypertension (5/26), proteinuria (2/26) and chronic renal insufficiency (CRI) (2/26). At the last follow-up (mean duration of follow-up: 76 months; range: 0.5-262 months), 19 children (mean age: 5.5 years) were asymptomatic, five (mean age: 8.5 years) had hypertension, two (mean age: 9.7 years) had proteinuria and two (mean age: 19 years) had CRI. Children presenting enlarged kidneys postnatally tended to have more clinical manifestations than their counterparts who did not. Of 25 siblings of the patients, seven had renal cysts; these were detected during childhood in five siblings and in utero in two siblings. In conclusion, prognosis is favourable in most children with prenatal ADPKD, at least during childhood. The sex of the transmitting parent is not a risk factor of prenatal ADPKD. A high proportion of siblings develop early renal cysts. Abnormalities visualized by ultrasonography appear to be associated to more clinical manifestations. PMID- 17124605 TI - Uranium induces oxidative stress in lung epithelial cells. AB - Uranium compounds are widely used in the nuclear fuel cycle, antitank weapons, tank armor, and also as a pigment to color ceramics and glass. Effective management of waste uranium compounds is necessary to prevent exposure to avoid adverse health effects on the population. Health risks associated with uranium exposure includes kidney disease and respiratory disorders. In addition, several published results have shown uranium or depleted uranium causes DNA damage, mutagenicity, cancer and neurological defects. In the current study, uranium toxicity was evaluated in rat lung epithelial cells. The study shows uranium induces significant oxidative stress in rat lung epithelial cells followed by concomitant decrease in the antioxidant potential of the cells. Treatment with uranium to rat lung epithelial cells also decreased cell proliferation after 72 h in culture. The decrease in cell proliferation was attributed to loss of total glutathione and superoxide dismutase in the presence of uranium. Thus the results indicate the ineffectiveness of antioxidant system's response to the oxidative stress induced by uranium in the cells. PMID- 17124607 TI - Expression of protein kinase C isoforms in cultured human retinal pigment epithelial cells. AB - BACKGROUND: Protein kinase C (PKC) is involved in both physiological and pathophysiological processes and plays an important role in signal transduction. The present studies were designed to examine the 12 isoforms (PKCalpha, PKCbetaI, PKCbetaII, PKCgamma, PKCdelta, PKCepsilon, PKCeta, PKCtheta, PKCmu, PKCxi, PKClambda and PKCiota) of PKC expressed in cultured human retinal pigment epithelium (RPE) cells. METHODS: Human RPE cells were investigated for 12 PKC isoforms at the mRNA, protein and cellular levels by reverse transcription (RT) PCR, Western blot analysis and laser scanning confocal microscope (LSCM), respectively. RESULTS: RT-PCR and Western blot analyses showed similar results for specific PKC isoforms in that both revealed that PKCalpha, PKCbetaI, PKCbetaII, PKCdelta, PKCepsilon, PKCtheta, PKCmu, PKCxi, PKClambda and PKCiota, but not PKCgamma and PKCeta, were constantly expressed in RPE cells, with the exception of PKCbetaI at the protein level. Confocal microscopy showed that ten PKC isoforms - PKCalpha, PKCbetaI, PKCbetaII, PKCdelta, PKCepsilon, PKCtheta, PKCxi, PKCiota, PKClambda and PKCmu - appeared almost exclusively in the cytoplasm of the cells. However, PKCgamma and PKCeta were not detected by staining. CONCLUSIONS: This study characterized the expression pattern of all 12 PKC isoforms and showed that ten of these (PKCalpha, PKCbetaI, PKCbetaII, PKCdelta, PKCepsilon, PKCtheta, PKCmu, PKCxi, PKClambda and PKCiota) are present in cultured human RPE cells. This identification provides the first step towards elucidating their roles in RPE cell proliferation. PMID- 17124606 TI - The right dose for every sex. AB - Sex chromosomes in different organisms are studied as model systems for chromatin regulation of transcription and epigenetics. Similar to the female X in mammals, the male X chromosome in Drosophila is involved in the process of dosage compensation. However, in contrast to one of the mammalian female X chromosomes undergoing inactivation, the Drosophila male X is transcriptionally upregulated by approximately twofold. The Drosophila male X is a remarkable example for a specialized, transcriptionally hyperactive chromatin domain that facilitates the study of chromatin regulation in the context of transcription, nuclear architecture, and chromatin remodeling. In addition, the rich phenomenology of dosage compensation in Drosophila provides an opportunity to explore the complexities of gene regulation through epigenetic chromatin configurations, histone modifications, and noncoding RNAs. Male-specific lethal (MSL) factors constitute the MSL complex or dosage compensation complex and are important for transcription regulation of X-linked genes. Recent biochemical studies have identified a number of interesting factors that associate with the MSL complex including components of the nuclear pore complex and exosome subunits. Furthermore, global analysis of MSL complex binding showed that MSL complexes are enriched on genes with preferential binding to 3' end of genes. Taken together, these findings suggest a role of the MSL complex in transcription elongation, RNA processing, and/or nuclear organization. PMID- 17124608 TI - Peroneal nerve damage by oblique proximal locking screw in tibial fracture nailing: a new emerging complication? AB - Interlocking intramedullary nailing currently is the preferred treatment for most tibial fractures requiring operative treatment. Good results with a relative low complication rate have been reported in large clinical series, as well as in comparative series. The reported incidence of neurological complications after tibial nailing varies, involving mainly the peroneal nerve. The mechanism of peroneal nerve damage in tibial fracture nailing is usually indirect, caused by leg traction or compartment syndrome. Direct peroneal nerve damage related to the proximal locking screw seems to be very rare since we were able to identify only one report in the English literature. We report a case of partial peroneal nerve damage caused by a long oblique proximal locking screw. Removal of the proximal locking screw leaded to a gradual improvement of the nerve function and a complete resolution at one year. This seems to be a new emerging iatrogenic complication related to nails designed with oblique proximal locking screws. We feel that the placement of the oblique proximal screw from medial to lateral side needs an extra care. Even fluoroscopy, does not give enough safety due to the spatial geometry of the proximal tibia and the known problems of viewing oblique interlocking screws with a two-dimensional image-intensifier. PMID- 17124609 TI - The effect of heme oxygenase-1 induction by glutamine on TNBS-induced colitis. The effect of glutamine on TNBS colitis. AB - BACKGROUND: Inflammatory bowel disease is a multifactorial inflammatory disease of the colon and rectum with an unknown etiology. In the present study, we aimed to investigate whether heme oxygenase-1 (HO-1) induction by glutamine could protect colitis-induced damage from oxidative, inflammatory, and apoptotic damage. METHOD: The rats were divided into four groups. Group 1 had TNBS colitis alone, group 2 had TNBS-induced colitis and glutamine 1 g/kg/day intragastric gavage for 3 days before TNBS solution administration and 15 days following TNBS solution administration, group 3 had glutamine alone 1 g/kg/day intragastric gavage for 18 days before being killed, and group 4 had isotonic saline solution alone 1 cm3/rat intragastric gavage for 18 days before being killed. Colonic malondialdehyde (MDA) levels, glutathione (GSH) levels, caspase-3 activities, and HO-1 expressions of the killed rats were measured. Nuclear factor kappa B (NF kappaB) and HO-1 expression were evaluated by immunohistochemical examination of the colonic tissue. RESULT: TNBS-induced colitis significantly increased the colonic MDA levels, caspase-3 activities, and HO-1 expression in comparison to the control group. Glutamine treatment was associated with increased HO-1 expression and GSH levels and decreased MDA levels and caspase-3 activity. Histopathological examination revealed that the intestinal mucosal structure was preserved in the glutamine-treated group. In addition to this, treatment with glutamine significantly increased HO-1 expression and decreased NF-kappaB expression by immunohistochemistry when compared to the TNBS-induced colitis group. CONCLUSION: Glutamine reduced colonic damage in TNBS-induced colitis. The mechanism of the protection associated with glutamine was due to antioxidant, antiapoptotic, anti-inflammatory, and HO-1 induction effects. PMID- 17124610 TI - The transcription factor Gcr1 stimulates cell growth by participating in nutrient responsive gene expression on a global level. AB - Transcriptomic reprogramming is critical to the coordination between growth and cell cycle progression in response to changing extracellular conditions. In Saccharomyces cerevisiae, the transcription factor Gcr1 contributes to this coordination by supporting maximum expression of G1 cyclins in addition to regulating both glucose-induced and glucose-repressed genes. We report here the comprehensive genome-wide expression profiling of gcr1Delta cells. Our data show that reduced expression of ribosomal protein genes in gcr1Delta cells is detectable both 20 min after glucose addition and in steady-state cultures of raffinose-grown cells, showing that this defect is not the result of slow growth or growth on a repressing sugar. However, the large cell phenotype of the gcr1Delta mutant occurs only in the presence of repressing sugars. GCR1 deletion also results in aberrant derepression of numerous glucose repressed loci; glucose grown gcr1Delta cells actively respire, demonstrating that this global alteration in transcription corresponds to significant changes at the physiological level. These data offer an insight into the coordination of growth and cell division by providing an integrated view of the transcriptomic, phenotypic, and metabolic consequences of GCR1 deletion. PMID- 17124611 TI - Dual-contrast single breath-hold 3D abdominal MR imaging. AB - OBJECT: Multiple contrasts are often helpful for a comprehensive diagnosis. In 3D abdominal MRI, breath-hold techniques are preferred for single contrast acquisitions to avoid respiratory artifacts. In this paper, highly accelerated parallel MRI is used to acquire large 3D abdominal volumes with two different contrasts within a single breath-hold. MATERIAL AND METHODS: In vivo studies have been performed on six healthy volunteers, combining T (1)- and T (2)-weighted, gradient- or spin-echo based scans, as well as water/fat resolved imaging in a single breath-hold. These 3D scans were acquired with an acceleration factor of six, using a prototype 32-element receive array. RESULTS: The presented approach was tested successfully on all volunteers. The whole liver area was covered by a FOV of 350 x 250 x 200 mm(3) for all scans with reasonable spatial resolution. Arbitrary scan protocols generating different contrasts have been shown to be combinable in this single breath-hold approach. Good spatial correspondence with negligible spatial offset was achieved for all different scan combinations acquired in overall breath-hold times between 15 and 25 s. CONCLUSION: Enabled by highly parallel imaging technology, this study demonstrates the technical feasibility and the promising image quality of single breath-hold dual contrast MRI. PMID- 17124612 TI - The use of intraoperative monitoring and treatment of symptomatic microemboli in carotid artery stenting: case report and discussion. AB - Carotid artery stenting is a recently introduced treatment in symptomatic atherosclerotic carotid artery disease with acceptable complication rates. The major risk is perioperative embolic stroke. Transcranial Doppler ultrasonography (TCD) can be used to identify embolic signals and guide therapy. We present a case of symptomatic embolization in a 72-year-old female following carotid stent deployment complicated by haemodynamic changes. Despite concurrent dual antiplatelet medication significant symptomatic embolization occurred even after restoration of the blood pressure, and modulation of the rate of embolization was achieved using dextran-40 guided by TCD monitoring. The patient recovered from an initially profound hemiparesis and dysphasia to minor sensory changes. Microemboli are common following carotid artery stenting and there appears to be a threshold phenomenon associated with prolonged embolization and progression to cerebral infarction. TCD can be used to detect particulate microemboli and therefore may be useful in guiding antithrombotic therapy in this setting. Dextran-40 has been shown to reduce the embolic load following carotid endarterectomy and was used to good effect in this patient in terms of both embolic load and clinical outcome. This is the first case of embolization following carotid stenting successfully treated with dextran-40, and offers a further option for therapeutic intervention in microembolism detected by TCD and stresses the importance of perioperative monitoring of embolic load for postoperative stroke risk. PMID- 17124613 TI - Adhesion molecules in pediatric intensive care patients with organ dysfunction syndrome. AB - OBJECTIVE: To determine serum concentrations of the soluble forms of vascular cell adhesion molecule 1 (VCAM-1), intracellular adhesion molecule 1 (ICAM-1), and E-selectin in ventilated neonatal and pediatric intensive care patients with varying severity of multiorgan dysfunction syndrome (MODS) with or without infection-triggered organ failure. DESIGN AND SETTING: Prospective pilot study, a level III neonatal and pediatric intensive care unit at a University children's Hospital. PATIENTS: We studied 22 ventilated pediatric (n=15) and neonatal (n=7) intensive care patients (aged 3 days-16 years). Inclusion criteria were mechanical ventilation and signs of at least one additional organ dysfunction (cardiovascular, respiratory, neurological, hematological, or renal). MEASUREMENTS AND RESULTS: Serum concentrations of the adhesion molecules were analyzed on the day of maximum organ dysfunction score and were quantitated by a sandwich ELISA technique. The overall mortality rate was 36% (8/22). Dysfunction of three or more organ systems was defined as MODS and was associated with a significant increase in VCAM-1 serum levels relative to dysfunction of three or fewer organ systems [median 1239 ng/ml (IQR 928-1615) vs. 766 ng/ml (644-915)]. A significant difference in E-selectin serum levels was found between organ failure of infectious (median 131 ng/ml, IQR 112-146) and noninfectious origin (68 ng/ml 49-105). CONCLUSIONS: Determination of adhesion molecules in pediatric intensive care patients raises the possibility of more specific pathophysiological understanding. E-selectin showed significantly different serum levels between infectious and noninfectious causes of organ failure. PMID- 17124614 TI - [Spinal infections]. AB - Regardless of the compartment involved, imaging of spinal infections and other spinal inflammatory diseases must be performed by magnetic resonance imaging (MRI). This allows early detection of changes in disk space or vertebral bodies. Associated paraspinal and epidural masses are depicted. However, in spinal cord inflammation MRI has a very limited specificity and cannot usually differentiate between the wide array of possible causes of intramedullary lesions. This article covers a spectrum of inflammatory lesions of the spinal cord. PMID- 17124615 TI - [Retained foreign bodies from the view of the OR nurse]. AB - Patient safety in the operating room is of high priority for the surgical team and for all healthcare workers involved. Counting procedures contribute to minimizing potential risks for the patients, especially in preventing them from leaving the operating room with retained, unintended foreign bodies. A total of 64% of operating room nurses involved in a survey are reported as having been involved in a surgical procedure in which the correct count prevented foreign bodies from being left in the patient. In addition, the strengths and potential weak points of the counting procedures could be identified. The results of the survey and drafts for a standard counting control as well as procedural recommendations are presented for discussion. A recommendation can be provided for each surgical department in order to check their own counting methods for avoiding potential risks. PMID- 17124616 TI - [Functional and aesthetic refinements of free flap coverage at the dorsum of the hand and distal forearm]. AB - INTRODUCTION: Free coverage of exposed functional structures of the hand and distal forearm can be achieved using a variety of free flaps. However, there is a lack of data in the literature which tissue components are best used for defect coverage of this specific area regarding aspects such as tendon gliding and tissue elasticity. The purpose of this retrospective study was to compare and to evaluate the functional and aesthetical results using free cutaneous, fascial and muscle flaps. PATIENTS AND METHODS: Between 1994 and 2002, a total of 56 patients underwent free flap coverage at the dorsal side of the hand and forearm at our institution. Of these patients, 20 with 21 free flaps met the study criteria and were available for follow-up examination. Depending on the tissue component used patients were allocated into three different groups. In group 1 eight patients received a fascial flap, in group 2 eight a cutaneous flap and in group 3 five a muscle flap. The mean follow-up was 50 months (range: 4-103 months). At follow-up examination the patients answered the Client Satisfaction Questionnaire (CSQ 8) and the DASH questionnaire. RESULTS: Regarding range of motion of the wrist and fingers, fascial flaps clearly showed the best results. Concerning grip strength and pinch grip, fascial and cutaneous flaps demonstrated the same results. Patients with cutaneous flaps showed the best DASH score. Regarding the overall aesthetical outcome, fascial and cutaneous flaps were far better than muscle flaps. Donor site morbidity was lowest for fascial flaps followed by muscle flaps and was highest for cutaneous flaps. CONCLUSION: Cutaneous and fascial flaps had the best functional and aesthetical results. Fascial flaps achieved the best aesthetical outcome of the donor site. We recommend free fascial flaps and cutaneous flaps as the first choice due to their advantages in the specific area of the dorsal site of the hand and distal forearm which requires pliable and thin tissue coverage. PMID- 17124617 TI - Introducing mesoscopic information into constitutive equations for arterial walls. AB - We propose a new elastic constitutive law for arterial tissue in which the limiting polymeric chain extensibility of both collagen and elastin fibres is accounted for. The elastic strain-energy function is separated additively into two parts: an isotropic contribution associated with the matrix (incorporating the elastin fibre network) and an anisotropic one associated with the collagen fibres. Information on the limiting extensibility in each case provides some mesoscopic input into the model. The (logarithm-based) model is compared with the Fung-Demiray exponential model and certain other recently proposed models. Some aspects of the elastic response under extension and inflation of a thin-walled circular cylindrical tube (the artery) are then examined and compared with the corresponding response of a rubber-like tube. We point out that our model, when both isotropic and anisotropic terms are included, can be developed to accommodate changing mechanical properties associated with degradation of the elastin and collagen by considering the material constants that define the limit of chain extensibility to evolve in time. PMID- 17124618 TI - Selection of heart failure patients for cardiac resynchronisation therapy: a role for PET? PMID- 17124619 TI - Issues in quantification of cardiac PET studies. PMID- 17124620 TI - Acute and chronic tryptophan depletion differentially regulate central 5-HT1A and 5-HT 2A receptor binding in the rat. AB - RATIONALE: Tryptophan depletion is used to reduce central serotonergic function and to investigate its role in psychiatric illness. Despite widespread clinical use, its effects on serotonin (5-HT) receptors have not been well characterized. OBJECTIVE: The aim of this study was to examine the effect of acute (ATD) and chronic tryptophan depletion (CTD) on free-plasma tryptophan (TRP), central TRP and 5-HT and brain 5-HT(1A) and 5-HT(2A) receptor binding in the rat. METHODS: TRP and 5-HT were measured by high-performance liquid chromatography and receptor levels determined by homogenate radioligand binding and in-vitro receptor autoradiography. RESULTS: Free-plasma TRP, central TRP and central 5-HT levels were significantly and similarly reduced by ATD and 1- and 3-week CTD compared to controls. ATD significantly reduced 5-HT(1A) binding in the dorsal raphe (14%) but did not significantly alter postsynaptic 5-HT(1A) binding (frontal cortex, remaining cortex and hippocampus) or 5-HT(2A) binding (cortex and striatum). One week CTD did not significantly alter cortical 5-HT(2A) binding or postsynaptic 5 HT(1A) binding. Furthermore, 3-week CTD did not significantly alter 5-HT(1A) binding but significantly increased cortical 5-HT(2A) binding without affecting striatal or hippocampal levels. In the CTD 1 and 3-week groups, rat body weight was significantly decreased as compared to controls. However, weight loss was not a confounding factor for decreased cortical 5-HT(2A)-receptor binding. CONCLUSION: ATD-induced reduction in somatodendritic 5-HT(1A) autoreceptor binding may represent an intrinsic 'homeostatic response' reducing serotonergic feedback in dorsal raphe projection areas. In contrast, the increase in 5-HT(2A) receptor after CTD may be a compensatory response to a long-term reduction in 5 HT. PMID- 17124621 TI - The role of 5-HT1a and 5-HT2a receptors in attention and motor control: a mechanistic study in healthy volunteers. AB - RATIONALE: Various studies have demonstrated a modulating role for serotonin in attention. Selective serotonin inhibitors have repeatedly been shown to impair performance in sustained attention tasks. OBJECTIVES: To assess the contribution of serotonin reuptake inhibition and specific blockade of the pre-synaptic 5 HT(1a) receptor and the 5-HT(2a) receptor to deficits in attention. MATERIALS AND METHODS: The study was conducted according to a randomized, double-blind, placebo controlled, four-way crossover design including 16 healthy volunteers. Treatments consisted of oral administration of the selective serotonin reuptake inhibitor (SSRI) escitalopram 20 mg + placebo; escitalopram 20 mg + ketanserin (5-HT(2a) antagonist), 50 mg; escitalopram 20 mg + pindolol (5-HT(1a) antagonist) 10 mg; and placebo + placebo on four separate days. A range of performance tasks were conducted to assess the subjects' attention and motor functions. RESULTS: Escitalopram administered alone impaired tracking performance in a divided attention task. The combination of escitalopram and pindolol and escitalopram and ketanserin impaired divided attention as compared to placebo. In addition, escitalopram and ketanserin impaired sustained attention. Divided attention impairment observed after combined treatments did not significantly differ from impairments after escitalopram alone. Sustained attention impairment observed after combined escitalopram and ketanserin significantly differed from escitalopram alone. CONCLUSIONS: 5HT(1a) blockade hardly affected SSRI effects on attention. Additional 5HT(2a) blockade, however, produced impairments of sustained attention and motor impulse control. PMID- 17124622 TI - Treatment of detrusor sphincter dyssynergia with baclofen and doxazosin. AB - Detrusor sphincter dyssynergia (DSD) is an involuntary contraction of the external urethral sphincter during detrusor contraction. A high proportion of patients needing repeat surgery and long term failure have both been described in the literature. In the present study, we evaluated clinical characteristics, underlying disorders and outcomes of conservative medical treatment in 21 female patients. Two patients were newly diagnosed multiple sclerosis. Urodynamic studies were performed in all symptomatic patients, and consisted of measurement of post-micturition residuals, urethral pressure profilometry and EMG cystometry according to the criteria of the International Continence Society. All patients were treated with baclofen 15 mg/day and doxazosin 4 mg/day. Seven patients received tolterodine 4 mg/day in addition to baclofen and doxazocin because they had detrusor hyperreflexia (DH). In conclusion, treatment with either combined baclofen and doxazosin or anticholinergic agent tolterodine appeared to be effective. In addition, it should be kept in mind that DSD could be the first sign to any neurologic diseases. PMID- 17124623 TI - Renoprotective effects of tea catechin in streptozotocin- induced diabetic rats. AB - Tea catechins, a class of flavonoids, are suggested to have biological effects, possibly mediated through their antioxidative properties. Recent data indicated that tea catechins suppressed proliferative changes in glomeruli and inhibited the development of glomerulosclerosis in partially nephrectomized rats. We thus sought to determine whether tea catechins may protect against renal dysfunction in streptozotocin-induced diabetic rats. Four groups of male Sprague-Dawley rats (n=11-15 per group), with and without streptozotocin-induced diabetes, were treated with and without catechins (5 mg/day) administered in the drinking water for 12 weeks. At the end of the treatment period, 24-hour urinary albumin excretion rate (AER), serum lipid peroxides as thiobarbituric acid reactive substrates (TBARS) and blood pressure were measured. Renal glomerular volume and interstitial fibrosis were assessed morphologically. Albuminuria developed progressively in untreated diabetic rats, resulting in a mean AER of 559+/-124 (mean+/-SE) versus 63+/-7 microg/day/100 g body weight in non-diabetic rats at 12 weeks (P<0.001). Catechin treatment significantly reduced AER to 287+/-56 microg/day/100 g body weight in diabetic rats (P=0.017 versus untreated diabetic rats). Increased interstitial fibrosis in the kidney, observed in untreated diabetic rats, was completely normalized with catechin treatment. Serum levels of TBARS and blood pressure were comparable among the four groups. In conclusion, administration of tea catechin retards the progression of functional and morphological changes in the kidney of streptozotocin-induced diabetic rats. PMID- 17124624 TI - The management of female urinary retention. AB - Female acute urinary retention (AUR) is relatively uncommon and often poorly managed. There are several common precipitants though much of the literature refers to female AUR as a psychogenic condition. The underlying abnormality is often detrusor failure, not outlet obstruction. Investigations should focus on identifying serious or reversible causes and should include a detailed history and physical examination, urine dipstick, culture and pelvic ultrasound. Patients should be catheterised and reversible causes should be treated. Women who fail to void after catheter removal should be taught ISC. Alpha-blockers are no better than placebo in the treatment of female AUR. There is no role for urethral dilatation. Patients with apparently idiopathic retention should be referred to a urologist with an interest in bladder dysfunction for consideration of urodynamics. PMID- 17124625 TI - Ureterosciatic hernia with compression of the sciatic nerve. AB - Ureterosciatic herniation is a rare cause of ureteral obstruction. Sciatic hernia is a well-defined anatomic defect that is the result of atrophy or abnormal development of piriform muscle. Patients with sciaitic hernias commonly present with symptoms of flank, abdominal, pelvic, lower back or thigh pain. The hernia sack can contain small bowel, ureter, ovary, colon or bladder. Ureterosciatic hernia causing ureteral obstruction should be surgical repaired. PMID- 17124626 TI - A gentle reminder in the laparoscopic era left-sided inferior vena cava. PMID- 17124627 TI - Understanding the experience of being taught by peers: the value of social and cognitive congruence. AB - BACKGROUND: Medical schools use supplemental peer-teaching programs even though there is little research on students' actual experiences with this form of instruction. PURPOSE: To understand the student experience of being taught by peers instead of by faculty. METHODS: We conducted focus groups with first- and second-year medical students participating in a supplemental peer-teaching program at one institution. From the learner focus group themes, we developed a questionnaire and surveyed all first-year students. RESULTS: Focus groups revealed four learner themes: learning from near-peers, exposure to second-year students, need for review and synthesis, teaching modalities and for the peer teachers, the theme of benefits for the teacher. Factor analysis of the survey responses resulted in three factors: second-year students as teachers, the benefit of peer-teachers instead of faculty, and the peer-teaching process. Scores on these factors correlated with attendance in the peer-teaching program (P < .05). CONCLUSIONS: Students valued learning from near-peers because of their recent experience with the materials and their ability to understand the students' struggles in medical school. Students with the highest participation in the program valued the unique aspects of this kind of teaching most. Areas for improvement for this program were identified. PMID- 17124628 TI - Molecular dynamics simulation of the human adenosine A3 receptor: agonist induced conformational changes of Trp243. AB - The adenosine A(3) receptor together with rhodopsin belongs to Class A of the G protein coupled receptors. As the crystal structure of bovine rhodopsin represents the dark (inactive) state of the receptor, the details of GPCR activation are still unknown. In this molecular dynamics study we investigate how the homology model of the human adenosine A(3) receptor responds to ligand exposure. To this end we placed the homology model in a POPC membrane model. After equilibrating for 13 ns an agonist (Cl-IB-MECA) and an inverse agonist (PSB 10) were placed inside the putative binding pocket. In the following 10 ns molecular dynamics simulation we observed a different behaviour of the side-chain torsions of Trp243(6.48), depending on the presence or absence of the agonist or inverse agonist. This conformational change of Trp243 correlates with the assumed influence of ligands on receptor activation. Other predicted conformational changes of the receptor could not be observed yet. So Trp243 may represent the first switch in receptor activation. PMID- 17124629 TI - PHASE: a new engine for pharmacophore perception, 3D QSAR model development, and 3D database screening: 1. Methodology and preliminary results. AB - We introduce PHASE, a highly flexible system for common pharmacophore identification and assessment, 3D QSAR model development, and 3D database creation and searching. The primary workflows and tasks supported by PHASE are described, and details of the underlying scientific methodologies are provided. Using results from previously published investigations, PHASE is compared directly to other ligand-based software for its ability to identify target pharmacophores, rationalize structure-activity data, and predict activities of external compounds. PMID- 17124630 TI - Conformational specificity of non-canonical base pairs and higher order structures in nucleic acids: crystal structure database analysis. AB - Non-canonical base pairs contribute immensely to the structural and functional variability of RNA, which calls for a detailed characterization of their spatial conformation. Intra-base pair parameters, namely propeller, buckle, open-angle, stagger, shear and stretch describe structure of base pairs indicating planarity and proximity of association between the two bases. In order to study the conformational specificities of non-canonical base pairs occurring in RNA crystal structures, we have upgraded NUPARM software to calculate these intra-base pair parameters using a new base pairing edge specific axis system. Analysis of base pairs and base triples with the new edge specific axis system indicate the presence of specific structural signatures for different classes of non-canonical pairs and triples. Differentiating features could be identified for pairs in cis or trans orientation, as well as those involving sugar edges or C-H-mediated hydrogen bonds. It was seen that propeller for all types of base pairs in cis orientation are generally negative, while those for trans base pairs do not have any preference. Formation of a base triple is seen to reduce propeller of the associated base pair along with reduction of overall flexibility of the pairs. We noticed that base pairs involving sugar edge are generally more non-planar, with large propeller or buckle values, presumably to avoid steric clash between the bulky sugar moieties. These specific conformational signatures often provide an insight into their role in the structural and functional context of RNA. PMID- 17124631 TI - Molecular modeling and bioinformatical analysis of the antibacterial target enzyme MurA from a drug design perspective. AB - The enzyme MurA (UDP-N-acetylglucosamine enolpyruvyl transferase) catalyzes the first cytoplasmatic step in the synthesis of murein precursors. This function is of vital relevance for bacteria, and the enzyme therefore represents an important target protein for the development of novel antibacterial compounds. Several X ray structures of liganded and un-liganded MurA have been published, which may be used for rational drug design. MurA, however, contains a highly flexible surface loop, which is involved in substrate and inhibitor binding. In the available X ray structures, the conformation of this surface loop varies, depending on the presence or absence of ligands or substrate and probably also on the crystal packing. The uncertainty of the low-energy, or "resting state" conformation of this surface loop hampers the application of rational drug design to this class of enzymes. We have therefore performed an extensive molecular dynamics study of the enzyme in order to identify one or several low-energy conformers. The results indicate that, at least in some of the X-ray structures, the conformation of the flexible surface loop is influenced by crystallographic contacts. Furthermore, three partially helical foldamers of the surface loop are identified which may resemble the resting states of the enzyme or intermediate states that are "traversed" during the substrate binding process. Another, very important aspect for the development of novel antibacterial compounds is the inter- and intra species variability of the target structure. We present a comparison of MurA sequences from 163 organisms which were analyzed under the aspects of enzyme mechanism, structure and drug design. The results allow us to identify the most promising binding sites for inhibitor interaction, which are present in MurA enzymes of most species and are expected to be insusceptible to resistance inducing mutations. PMID- 17124632 TI - Microwave-assisted extraction of embelin from Embelia ribes. AB - A rapid and efficient microwave-assisted extraction (MAE) process for the selective extraction of embelin from Embelia ribes was developed. Solvent selection, microwave energy input and solid loading were optimized. The rate of extraction and purity of embelin depended upon the solvent used and exposure time to microwaves. Maximum MAE was achieved in acetone with total yield of 92% (w/w) embelin with 90% (w/w) purity with 1% (w/v) raw material loading at 150 W power level in 80 s. Non-polar solvents, such as hexane and dichloromethane, were not effective for the selective extraction of embelin. PMID- 17124633 TI - Subunit dissociation and stability alteration of D hydantoinase deleted at the terminal amino acid residue. AB - Two variants of D hydantoinase (HYD), created by deletion of one amino acid residue of at either the N- or C-terminus, were expressed in Escherichia coli and purified by two-step chromatography. Compared with HYD, HYDc1 with the C-terminal Arg deletion retained 43% activity, while HYDn1 with the N-terminal Ser deletion had no activity using DL Hydantoin as substrate. Based on HYD dimer with a molecular weight of 103 kDa, HYDc1 is a monomer of 52 kDa and HYDn1 is a mixture of dimer and monomer. Moreover, HYDc1 displayed higher pH stability and lower thermal stability compared to HYD. In addition, the secondary and tertiary structures of HYDc1 were not significantly changed in contrast to the ones of HYDn1. All data imply that the C-terminal Arg of the HYD is crucial for homodimeric architecture of the enzyme, but non-essential for catalysis, while the N-terminal Ser is required for both conformation and catalysis of the enzyme. PMID- 17124634 TI - Interstitial cystitis in a woman with systemic mastocytosis. AB - Studies have reported detrusor mastocytosis in patients with interstitial cystitis. The author describes a patient with systemic mastocytosis who was confirmed to have detrusor mastocytosis and interstitial cystitis. She responded to therapy with pentosanpolysulfate. The literature on systemic mastocytosis and the role of mast cells in the pathophysiology of interstitial cystitis are reviewed. PMID- 17124635 TI - Physioanatomical relationship of the external anal sphincter to the bulbocavernosus muscle in the female. AB - Both external anal sphincter (EAS) and bulbocavernosus muscle (BCM) have been shown anatomically and physiologically to constitute one muscle in males. We investigated the hypothesis that the EAS and BCM have similar anatomical pattern in females. The study consisted of cadaveric dissection, electromyographic recordings and inferior rectal nerve stimulation. Bulbocavernosus reflex action was performed in 16 healthy women before and after anesthetization of the EAS and BCM. The EAS extended forward across the perineal body and became continuous with the BCM in the labia majora. Glans clitoris (GC) or inferior rectal nerve stimulation effected synchronous EAS and BCM contractions with identical action potentials. GC stimulation while the EAS or BCM was anesthetized produced neither EAS nor BCM response. Similarly, stimulation of the anesthetized GC produced no EAS or BCM response. The BCM and EAS apparently constitute a single muscle, which seems to play dual and yet synchronous roles in fecal control and sexual response. PMID- 17124636 TI - The effects of combined treatment with niacin and chromium on the renal tissues of hyperlipidemic rats. AB - In this study, 12 months old female Swiss albino rats were used. They were randomly divided into four groups. The animals of group I were fed with pellet chow. Group II were fed with pellet chow and treated with 250 microg/kg CrCl3 x 6H2O and 100 mg/kg niacin for 45 days. Group III were fed a lipogenic diet consisting of 2% cholesterol, 0.5% cholic acid and 2% sunflower oil added to the pellet chow, and given 3%alcoholic water for 60 days. Group IV were fed with the same lipogenic diet for 60 day sand treated by gavage technique to rats at a dose of 250 micro/kg CrCl3 x 6H2O and 100 mg/kg niacin for 45 days, 15 days after experimental animals were rendered hyperlipidemic. At the 60th day, renal tissue and blood samples were taken from the animals. The sections were examined under light and electron microscopy. The degenerative changes were much more in the hyperlipidemic rats than the control group. The changes in renal tissue were also observed in hyperlipidemic animals given niacin and chromium. In the hyperlipidemic rats, renal glutathione levels decreased and renal lipid peroxidation levels, and serum urea and creatinine levels were increased. But, renal glutathione levels increased and lipid peroxidation levels and serum urea and creatinine levels decreased in hyperlipidemic rats given niacin and chromium. The purpose of this study was to investigate whether a protective effect of a combination of niacin and chromium is present on the renal tissue of hyperlipidemic rats or not. In conclusion, we can say that niacin and chromium do not have a protective effect on the morphology of the renal tissue of hyperlipidemic rats, except a protective effect on their biochemical parameters. PMID- 17124637 TI - Human epidermoid A431 cells express functional nicotinic acid receptor HM74a. AB - Nicotinic acid (niacin) has been used clinically to manage dyslipidemia for many years. The molecular target of nicotinic acid was unknown until the recent revelation of human G-coupled receptor HM74a as the high affinity receptor for nicotinic acid. In searching for a cell line expressing endogenous human HM74a receptor, we have identified that the A431 cell line, a human epidermoid cell line, expresses a high level of HM74a receptor. An HM74a-specific real time PCR probe set was designed and the mRNA levels of HM74a in A431 and 32 other cultured cell lines were measured quantitatively. When the mRNA expression of HM74a in A431 cells was compared to that in human primary preadipocytes, adipocytes and adipose tissue, we found that the level in A431 was about 10- fold higher than that in adipocytes and adipose tissue. The ratio of HM74a:HM74 mRNA was measured quantitatively and it was determined to be 3:2 in A431 cells. The function of the HM74a receptor in A431 cells was evaluated for its ability to inhibit forskolin induced cAMP production. Pertussis toxin treatment abolished the inhibition. Our data suggest that the A431 cell line may serve as a cellular model for further investigation of niacin/HM74a-mediated signal transduction in modulating metabolism. A431 cell line may also provide a valuable cell model to study prostaglandin production upon HM74a activation to improve our understanding of niacin/HM74a-mediated skin flushing. PMID- 17124638 TI - [Impact of structure and context on utilization of socio-psychiatric services in Saxony--a multi-level-analysis]. AB - OBJECTIVE: To analyse the influence of structural (service) aspects and context (catchment area) variables on the individual use of socio-psychiatric services in Saxony. METHOD: Data collected in 34 services were used. For the analysis of the variance components a multi-level model with random effects was computed using data at two levels ([1] client, and [2] service/catchment area). RESULTS: Service use was substantially influenced by structural/context factors which accounted for a proportion of 10 to 50 % of the variance depending on the (sub-)group of users included in the analysis. The impact of service structure and context on service use was particularly strong in the subgroup of service users with very high service utilisation. PMID- 17124639 TI - [Gabapentin treatment in a female patient with panic disorder and adverse effects under carbamazepine during benzodiazepine withdrawal]. AB - Despite their addictive potential, benzodiazepines belong to the most often prescribed drugs. We report on a patient with alprazolam dependence, who initially was treated with carbamazepine because of severe withdrawal symptoms. Due to liver enzyme elevation related to carbamazepine, we had to stop this treatment and instead of that started gabapentin treatment. Under this new therapy, the patient showed a dramatic relief of withdrawal symptoms and of the panic attacks recurring during withdrawal. Hence, due to their effectiveness and tolerability, newer anticonvulsants could be considered as medication for benzodiazepine withdrawal and as an alternative for benzodiazepine treatment in panic disorders. PMID- 17124640 TI - ECT anesthesia: the lighter the better? AB - BACKGROUND: Electroconvulsive therapy (ECT) is a most effective treatment for patients with major affective disorders. The influence of anesthetic drugs on seizure "adequacy" or on treatment success has not been systematically investigated. METHODS: A bispectral EEG index score (BIS) was used to identify the depth of anesthesia during ECT. Our study included 22 major depressive episode (MDE) patients expanding to 219 ECTs (05/05-01/06) with no limitations of concurrent medication. RESULTS: Fourteen out of the 22 patients showed full remission. Individual number of ECT sessions needed to reach full remission correlated negatively with mean pre-ECT BIS values (p=0.001). Additionally, using a repeated measurement regression analysis significant correlations were found for pre-ECT BIS versus motor response time, seizure concordance, ictal coherence and peak heart rate. CONCLUSION: The results of our study suggest BIS-levels as a predictor of faster ECT response. Controlling BIS-levels before stimulation may have an additional effect on treatment success. PMID- 17124641 TI - The effect of donepezil on sleep in elderly, healthy persons: a double-blind placebo-controlled study. AB - INTRODUCTION: Previous research in younger individuals has shown that acetylcholinesterase inhibitors tend to enhance REM sleep. METHODS: Forty-two healthy elderly persons participated in a double-blind placebo-controlled polysomnographic study (parallel group design). RESULTS: The present study indicates that in the elderly persons, donepezil, an acetylcholinesterase inhibitor also exerts a marked effect on REM sleep parameters: REM density was increased whereas REM latency was reduced, thus, confirming the findings of our pilot study described earlier. CONCLUSION: Whether the cholinergic stimulation measured by polysomnography is related to treatment efficacy is a very interesting but an open question. Based on the findings that REM sleep is associated with memory consolidation, the question whether REM sleep augmentation enhances memory performance-as suggested by the findings of the pilot study-seems to be an interesting topic for future research. PMID- 17124642 TI - Initial treatment of severe acute psychosis with fast orally disintegrating risperidone tablets. AB - INTRODUCTION: Although the use of atypical antipsychotics is the standard of care in the maintenance treatment of psychosis, most clinicians still rely on conventional neuroleptics to treat acutely agitated psychotic patients. The objective of this study was to evaluate the effectiveness and safety of a fast orally disintegrating tablet formulation of risperidone in the initial treatment of a large sample of very acutely ill psychotic patients. METHODS: In this multi center, prospective, open-label observational trial, 191 schizophrenic patients were treated upon admission to hospital with fast orally disintegrating risperidone tablets for up to seven days. Co-medication was per usual clinical practice and at physician's discretion. Psychopathology was rated at baseline, 2, 24 and 48 hours and 4 and 7 days after initiation of therapy. RESULTS: A mean PANSS total score of 114.3+/-23.4 at baseline reflected a severely exacerbated patient population. The PANSS total score was significantly reduced to 83.6+/ 26.8 (p<0.0001) and the CGI from 5.6+/-0.7 to 4.5+/-1.1 (p<0.0001) after 7 days. The median time to calmness was 70 min and the associated PANSS item 4 (excitation) dropped two hours after the first intake of the study medication from 4.3+/-1.5 to 3.1+/-1.5 (p<0.0001). A total of 172 patients (90.1%) out of 191 completed the study. The median risperidone dose was 2 mg/d at the initiation of therapy and 4 mg/d after one week. CONCLUSION: Oral treatment of acutely exacerbated schizophrenic patients with fast orally disintegrating risperidone tablets, alone or in combination with benzodiazepines, was associated with a rapid onset of action and a significant and clinically relevant improvement of acute symptoms. PMID- 17124643 TI - Comparison of Hypericum extract WS 5570 and paroxetine in ongoing treatment after recovery from an episode of moderate to severe depression: results from a randomized multicenter study. AB - OBJECTIVE: To test and compare the efficacy and safety of Hypericum extract WS 5570 to paroxetine, a potent SSRI, in patients suffering from moderate or severe depression according to DSM-IV criteria. METHODS: In a multicenter, randomized, double-blind phase III study, the changes in moderate to severe major depression DSM-IV; 17-item Hamilton Depression Rating Scale (HAM-D total>or=22) after an acute treatment with Hypericum extract WS 5570 or paroxetine were analyzed in a 16-week continuation phase for relapse prevention. Patients with a HAM-D total score decrease of >or=50% during the 6 weeks of acute treatment were asked to continue the treatment for another 4 months. One-hundred and thirty-three adult out-patients who received maintenance doses of 900 (n=33) or 1800 mg/d (n=38) of WS 5570 and 20 (n=28) or 40 mg/d (n=34) of paroxetine, respectively, were included. The relevant dosage was already fixed during the acute treatment. RESULTS: Between baseline of the acute phase and end of continuation treatment the HAM-D total score decreased from 25.3+/-2.5 (mean+/-SD) to 4.3+/-6.2 points for WS 5570 and from 25.3+/-2.6 to 5.2+/-5.5 points for paroxetine (p=0.49, two sided t-test; median relative decrease: 92.0 and 85.5%, respectively). During maintenance treatment alone (day 154-day 42), 61.6% of the patients randomized to WS 5570 and 54.6% treated with paroxetine showed an additional reduction (p=0.59) with respect to the HAM-D total score. Remission (HAM-D endpoint total score below 8) occurred in 81.6% (31 patients) of the patients for WS 5570 and in 71.4% (30 patients) for paroxetine (p=0.29). Three patients in the WS 5570 group and 2 patients in the paroxetine group showed a HAM-D increase>5 points during continuation treatment. In the continuation phase there were 0.006 adverse events per day of exposure for WS 5570 and 0.007 events for paroxetine. CONCLUSION: This study showed that WS 5570 and paroxetine were similarly effective in preventing relapse in a continuation treatment after recovery from an episode of moderate to severe depression and point therefore to an important alternative treatment option for long-term relapse-prevention. PMID- 17124644 TI - Apoptosis of CD4+ T and natural killer cells in Alzheimer's disease. AB - BACKGROUND: Immunotherapy appears to be a potent treatment against Alzheimer's disease (AD), but the mechanisms underlying neural-immune interaction are still not known. METHODS: Here, we determined cell death and distribution of lymphocyte subsets of peripheral blood mononuclear cells (PBMC) in AD and aging, e.g. T (CD4+ CD3+, CD8+ CD3+), B (CD19+) and NK (CD16++CD56+) cells. RESULTS: Increased apoptosis was found in CD4+ T and NK cells in AD, while in aging all subsets were affected. The expression of anti-apoptotic Bcl2 correlated with observed cell death in T-helper and B cells irrespective of dementia. The levels of Bcl2 in T cells were significantly increased in mild AD. Apoptosis and Bcl2 levels were also elevated in the APP (751SL)xPS1 (M146L) transgenic mouse model. CONCLUSION: The mechanisms triggering apoptosis and activation of lymphocytes in AD appear therefore to be different than those in immunosenescence and possibly bear an important biomarker to monitor immunotherapy in AD. PMID- 17124645 TI - Bupropion in atopic dermatitis. AB - Bupropion is an antidepressive drug whose main mechanism of action seems to be the inhibition of noradrenaline and dopamine recapture. We present a clinical case of a 50-year-old person with severe atopic dermatitis (A.D.) and without psychiatric symptoms associated, who was casually treated with bupropion with very good results. The mechanisms of action are not yet known, given the diversity of the implied physiopathology mechanisms: genetic, immunological, inflammatory, etc. The use of bupropion in serious and resistant cases to other treatments could constitute a pertinent therapeutic alternative. PMID- 17124646 TI - Pharmacokinetics and safety of combination therapy with lithium and risperidone. PMID- 17124647 TI - Successful treatment of alcohol withdrawal with trazodone. AB - Trazodone, a second generation antidepressant, is helpful in the treatment of lasting alcohol withdrawal symptoms, such as sleep disturbances and craving. We report a case suggesting that trazodone is also efficient for the treatment of the acute phase of alcohol withdrawal treatment. A 30-year-old male patient with severe alcohol withdrawal symptoms was successfully treated with an adjuvant administration of 600 mg trazodone per day, while he had remained symptomatic for several days under high doses of diazepam and clomethiazolum. Controlled trials with trazodone for this indication could be useful. PMID- 17124648 TI - Two cases of bipolar disorder successfully stabilized for five years with a low dose of risperidone and lithium. AB - Two patients with bipolar I disorder were successfully treated with a combination of risperidone and lithium in their acute manic states and maintenance periods. Although lithium monotherapy alone could not prevent relapse in these patients, the addition of a low dose of risperidone was well tolerated and effective for preventing recurrence over the long term. Plasma levels of HVA and MHPG were found to be elevated during the manic episodes and gradually decreased after the risperidone treatment. These results indicate that a low dose of risperidone to the lithium regimen was an effective and well tolerated means for treatment in the acute manic state and the later maintenance period in parallel with decreasing plasma levels of HVA and MHPG. PMID- 17124649 TI - Follow-up of a pregnancy with risperidone microspheres. AB - A follow-up of a case of pregnancy with risperidone microspheres is reported. Overall, there was a normal development of the female child during pregnancy and the first 2 1/2 years of her life. PMID- 17124650 TI - Cotrimoxazole-induced psychosis: a case report and review of literature. AB - We present a case of acute psychosis in a 46-year-old woman who had been treated orally with cotrimoxazole because of a severe infection of the urinary tract. She had started to develop psychotic symptoms with bizarre behavior two days before admission. Following discontinuation of antibiotic therapy, including cessation of treatment with cotrimoxazole and the induction of antipsychotic treatment, her mental state resolved to a stable premorbid level within 36 hours. PMID- 17124651 TI - Dyskinesia during treatment with duloxetine. PMID- 17124652 TI - [A human case of cyclosporiasis after traveling in the subtropics]. AB - In this study, Cyclospora oocysts were detected in a 64-year old man who complained of persistent diarrhea, abdominal pain, nausea and vomiting after visiting the Greek Islands in a sailing boat. Round oocysts about 8-9 microm in size with wrinkled walls that varied in color from red to pale pink after staining were found in Kinyoun's modified acid fast stained stool samples. Public health offices and laboratories, general practitioners, and medical microbiology labs should be informed that more attention should be paid to cyclosporiasis causing diarrheal illness and which requires specific screening methods with experienced microscopists in laboratories. PMID- 17124653 TI - [Malaria cases detected by active surveillance in Malatya]. AB - Malaria continues to be an important health problem in and around Malatya. Although the disease can be kept under control thanks to the regular work of the Centre for the Struggle against Malaria, it can not be eradicated fully due to the socio-economic conditions in the region. The aim of this study was to determine the prevalence of the disease through active surveillance in those regions where it is most common. From 2003, until 2004, 8856 blood samples were collected at one month intervals and 66 patients were found to be positive. Sixty three of the patients were Plasmodium vivax positive and 3 patients, P. falciparum, positive. The questionnaire administered to the positive patients (ranging from 02-70 years of age) revealed that 8 of the cases originated from extra-provincial sources. Men constitute 43% of all patients and women, 23%. Consequently it was concluded in this study that malaria threatens the people in the region. Therefore it was considered important for the Malatya Centre for the Struggle against Malaria and the local university to cooperate in the struggle against malaria by offering planned training programs in public health. PMID- 17124654 TI - [Evaluation of malaria cases in Manisa from 2002 to 2004]. AB - The province of Manisa is a relatively well developed agricultural and industrial center in western Turkey. There is a regular and extensive influx of workers from malaria-endemic regions of Anatolia to the province of Manisa during certain periods of the year, leading to contact between the local people and incoming workers. Major seasonal elevations can be detected in the prevalence of malaria in the province of Manisa and therefore an active fight against malaria is still carried out. In the present study, malaria cases detected and reported by the Malaria Control Dispensary of the Manisa City Health Centre during the last 3 years in Manisa have been evaluated. Comparison of present and prior data of malaria prevalence in Manisa has shown a remarkable decrease in recent years. A total of 14, 5 and 5 cases were reported in 2002, 2003 and 2004, respectively. Classification of the cases according to their origin (local or immigrant), age and the time diagnosis was also presented. PMID- 17124655 TI - [Seroprevalence of Neospora caninum in cattle in the region of Kayseri]. AB - This study was carried out in order to determine the seroprevalence of Neospora caninum in cattle in the region of Kayseri. Blood samples were collected from 186 cattle and sera were obtained from these samples. Sera samples were analyzed for antibodies against N. caninum using a commercially available competitive ELISA (c ELISA) kit. According to the ELISA test results, seroprevalence of N. caninum in the region of Kayseri was found to be 7%. Three of 9 aborting cows (33.3%) were found to be seropositive. The seropositivity differences between aborting and non aborting cows were statistically significant (p < 0.05). PMID- 17124656 TI - [Parasitic fauna and the frequency of Entamoeba histolytica/Entamoeba dispar detected by ELISA in stool samples in Sanliurfa, Turkey]. AB - Amoebiasis is a significant health problem in developing countries. Humans are infected by two morphologically identical species of Entamoeba. Entamoeba histolytica causes amebic colitis and liver abscess, and Entamoeba dispar is noninvasive. The aim of this study was to determine the prevalence of the E. histolytica/E. dispar complex using the ELISA method on stools of patients. A total of 1600 stool specimens were examined using Lugol preparations and the modified Ritchie method. A total of 583 (36.4%) of the stool specimens were found to be positive for one or more than one parasite. Twenty two subjects (3.8%) of the study population with intestinal parasites harbored two parasites and one subject (0.2%), three parasites. A total of 87 stool specimens that were doubtful using the Lugol method were examined by the E. histolytica specific sensu-lato antigen based ELISA test and the trichrome staining method. Of these 87 specimens, 23 (26.4%) specimens were positive for E. histolytica/E. dispar trophozoites/cysts microscopically using trichrome staining and 19 (21.7%) of the stool specimens were found to be positive for the E. histolytica/E. dispar complex by the ELISA test. PMID- 17124657 TI - [The prevalence of Enterobius vermicularis in schoolchildren and affecting factors]. AB - This study was carried out in order to determine the factors affecting the presence of Enterobius vermicularis in schoolchildren. This investigation was a cross-sectional and analytic study. The dependent variable was the presence of parasites and the independent variables were the characteristics of the children, families, houses, toilets and drinking water. A stool specimen was taken in the morning using the cellophane tape method, and examined the same day by microscopy. Data were evaluated using the Chi square test and logistic regression analysis and p < 0.05 was accepted as being statistically significant. There were 529 students in the kindergarten and 1-5 classes in the Isikkent and Sait Guzelcan primary schools in the region of the Isikkent Health Center in Izmir. However specimens could only be obtained from 477 (90.2%) students. The mean age of children was 8.6+/-2.0 years. The number of residents in their homes averaged 5.4+/-2.0, the average number of children in the homes was 3.2+/-1.9 and 290 (60.8%) houses were single dwellings. The source for piped water in 404 houses (84.7%) was the city network, and there were modern sanitary facilities (toilets) in 377 (70.6%) houses. Enterobius vermicularis was found in 209 (43.8%) children. According to logistic regression analyses, the rate of Enterobius vermicularis was found to be 3.05 times higher in students from the Sait Guzelcan primary school, if there were more than 6 residents in the home 2.05 times even higher and 2.02 times still higher if there were no sanitary facilities at his/her home. When there was a history of parasites in the family, the risk was significantly decreased. The prevalence of Enterobius vermicularis in schoolchildren was higher in slum areas, in crowded homes and in those that lacked modern sanitary facilities. PMID- 17124658 TI - [Prevalence of helminths in dogs in the region of Van and their potential public health significance]. AB - This study was performed in order to determine the prevalence of helminth infections among dogs and to emphasize the importance of this concept for public health. All selected dogs were from the city of Van and a total of 115 fecal samples were examined. Of these, 61 were from stray dogs, 26 were breeders and the final 28 dogs were dogs belonging to institutions. All samples were examined in the laboratory of the parasitology laboratory of the Faculty of Veterinary Medicine, Yuzuncu Yil University. Fulleborn's flotation method was used. Sixty nine (60.0%) of the dogs were found to be infected by one or more helminth species. Two Cestoda species and 4 Nematoda species were detected. Toxascaris leonina (23.5%), Taenia spp. (14.8%), Ancylostoma caninum (8.7%), Dipylidium caninum (3.5%), Uncinaria stenocephala (0.9%) and Toxocara canis (13.9%) were responsible for infections. Although the rates of infection between stray (59.0%) and official dogs (46.4%) was not significant, the rate was statistically significant according to Duncan's t-test ( < 0.05) in breeder dogs as compared to the previous 2 groups. The results show that helminth infection in dogs in Van is very high. The high prevalence of T. canis, Taenia spp., and A. caninum in dogs in Van is considered to be critical from the view point of public health importance. In conclusion, helminth infection of dogs is still a very serious problem for public health in Van. PMID- 17124659 TI - [Acute hepatitis cysticercosa and pneumonitis cysticercosa in a lamb: case report]. AB - The diseases known as hepatitis cysticercosa and pneumonitis cysticercosa are caused by migrating Cysticercus tenuicollis, the intermediate stage of Taenia hydatigena, in the liver and lung tissue of intermediate hosts like sheep, goats, cattle, pigs and squirrels. In this study, parasitological and pathological examinations were performed on the liver and lung tissue samples of a one month old lamb that had died in a herd of Karadayi village, Bunyan, Kayseri. After the examination, hepatitis cysticercosa and parasitic verminous pneumonia (Pneumonitis cysticercosa) were diagnosed and the results were discussed. PMID- 17124660 TI - [The prevalence of Pediculus capitis in students of eight primary schools in the rural area of the Kayseri province]. AB - This study was performed, from May-June 2005, on 648 (51.4%) males and 613 (48.6%) females making total of 1261 students from eight primary schools in following towns, Yemliha, Mahzemin, Gesi, Gunesli, Cirgalan, and Agirnas, in the rural area of the Kayseri Province. Either the adult or egg forms of Pediculus humanus var. capitis were found in 16 (2.1%) male students and 101 (16.4%), female making a total of 117 (9.1%) students. The prevalence of infestation was significantly higher in girls than in boys, the difference was found statistically significant (chi2=71.77, p < 0.05). In these eight primary schools the prevalence rate of infestation was also found statistically significant (chi 2= 95.7, p < 0.05). In conclusion, Pediculus capitis is still a public health problem. For an effective fight against this disease, students, teachers and parents must be acquainted with this subject and also surveillance must be increased. PMID- 17124661 TI - [Investigation of the ecological characteristics of the malaria vector Anopheles claviger (Diptera: Culicidae) in Sanliurfa (Siverek)]. AB - In this study, the seasonal larval and adult population dynamics, the seasonal gonotrophic status and feeding activity of Anopheles claviger were investigated from June 2000 to August 2002 in the Siverek district of the Sanliurfa Province. Monthly samples were collected using light, animal, and human traps as well as the mouth aspirator in the study of adult populations and the dipper methods in larval populations. At the end of the study, it was found that the adult population was reproductively active between June and September. Reproductive activity came to an end in the beginning of October and by November the adult population had totally disappeared so that the population will be carried over to the next season by larval over-wintering between November and April. Unfed, fed and gravid adults were sampled continuously from June to September in stable quadrates and Anopheles claviger was shown to display endophagic feeding and endophilic resting behavior. Results obtained from animal and human traps showed that adult feeding activity increased during sunrise and sunset hours. Anthropophilic tendency in the population was determined to be 18.78% and 14.22% in 2000 and 2001, respectively. PMID- 17124662 TI - [Culicoides latr, 1809 (Diptera: Ceratopogonidae) species in the Nigde province]. AB - This study was carried out in order to detect Culicoides species in the Nigde province (Tepe and Mezbaha) from June 2001-October 2001. Light traps were used for the collection of Culicoides specimens. The Culicoides specimens were cleared in phenol alcohol and they were mounted on slides. In this study a total of 593 Culicoides specimens composed of 548 females and 45 males were collected and 12 species (Culicoides punctatus, C. circumscriptus, C. festivipennis, C. gejgelensis, C. longipennis, C. maritimus, C. picturatus, C. puncticollis, C. nubeculosus, C. odiatus, C. pumilus and C. subneglectus), were identified. PMID- 17124663 TI - [Cuclotogaster heterographus (Mallophaga: Lipeuridae) infestation on ring-necked pheasants (Phasianus colchicus) in Konya]. AB - The aim of this study was to detect ectoparasites on ring-necked pheasants (Phasianus colchicus) in Konya. On the Research and Experimental Farm of the Faculty of Veterinary Medicine at Selcuk University, 55 ring-necked pheasants were observed to be infested with ectoparasites. Because of this, the pheasants were put into a large cartoon-box, the bottom of which was covered with a white paper and were treated with propoxur for a few minutes. The ectoparasites that had fallen on the paper were transferred to a tube containing 70% ethyl alcohol. After being cleared in lactophenol for a few days, they were mounted on slides in Faure-Forte medium. All of the ectoparasites were identified as the chewing lice (Mallophaga), Cuclotogaster heterographus. This is the first study that has recorded the presence of C. heterographus on ring-necked pheasants in Turkey. PMID- 17124664 TI - The first recording of Piagetiella titan (Menoponidae: Mallophaga) on a white pelican (Pelecanus onocrotalus, Linneaus) in Turkey. AB - During the examination of a wounded white pelican (Pelecanus onocrotalus) brought to the clinic of Faculty of Veterinary Medicine, Selcuk University, lice infestation of the oral cavity was observed and 26 lice were collected. The lice were identified as Piagetiella titan during microscopical examination. No other report of this particular subject in Turkey was found. Therefore, this original observation is reported as the first case of P. titan infestation of a white pelican found in Turkey. This report is being included in the data of the parasitic fauna of Turkey. PMID- 17124665 TI - [Cavicola myiasis caused by many Oestrus ovis (Linnaeus, 1761, Diptera: Oestridae) larvae in a sheep]. AB - When the head of a sheep was split into two along its longitudinal axis during a pathological examination, a large number of Oestrus ovis larvae were found in the nasal cavities, middle nasal meatus and sinuses of the sheep. A total of 114 Oestrus ovis larvae were collected, including 87 (76.32%) first stage larvae, 25 (21.93%) second stage larvae and 2 (1.75%) third stage larvae from the regions given above. PMID- 17124666 TI - [Allergic diseases and parasitosis]. AB - During recent decades, studies from Western countries suggest that there has been a significant increase in the incidence and prevalence of atopic diseases. Epidemiological studies have shown an inverse relationship between geohelminth infection and allergy leading to the suggestion that geohelminths protect against allergy. Geohelminth parasites modulate allergic inflammation directed against parasite antigens and the same mechanisms may affect responses to inhalant aeroallergens. Recently, it has been demonstrated that helminths induce suppressed host immune responses by the priming for regulatory T cells. New treatments being considered for the treatment of asthma include live infections with hookworms. Insights provided by how geohelminths modulate inflammatory responses may allow the development of new treatments that mimic these effects. PMID- 17124667 TI - [Metazoan parasites of carp (Cyprinus carpio Linnaeus, 1758) and vimba (Vimba vimba Linnaeus, 1758) in the Sapanca lake]. AB - In this study metazoan parasites of carp (Cyprinus carpio Linnaeus, 1758) and vimba (Vimba vimba Linnaeus, 1758) in the Lake Sapanca were investigated from January 2004 to December 2004. A total of 31 fish were investigated. Out of the 16 C. carpio, 13 were infected by parasites and of the 15 V. vimba, 11. The parasites found in C. carpio were: Dactylogyrus phoxini (Malevitskaya, 1949), Dactylogyrus extensus (Muller, Van Cleave, 1932), Gyrodactylus sp. Monogenoidea, Caryophyllaeus laticeps (Pallas, 1781), Bothriocephalus acheilognathi (Rud, 1808) Cestoidea, Diplostomum sp. Trematoda, Glochidium sp. Bivalvia. The parasites found in V. vimba were: Dactylogyrus sphyrna (Linstow, 1878), Dactylogyrus cornu (Linstow, 1878), Dactylogyrus cornoides (Glaser et Gussev, 1971) Monogenoidea, Aspidogaster limacoides (Dies., 1835), Posthodiplostomum cuticola (Nordmann, 1832), Tylodelphys clavata (Nordmann, 1832), Diplostomum sp., Tetracotyle sp. (Linstow, 1856) Trematoda, Neoechinorhynchus rutili (Muller, 1780) Acanthocephala, Glochidium sp. Bivalvia, Argulus foliaceus (Linnaeus, 1758), Ergasilus sieboldi (Nordmann, 1832) Crustacea. The parasites were investigated according to the place they were found in the host, prevalence and intensity of infections in the fish. Also the average intensity of the infection as well as the minimum and maximum intensities was estimated. PMID- 17124668 TI - [Occurrence of helminths during spring and autumn months on rudd (Scardinus erythrophthalmus L. 1758) from Kocadere stream (Bursa)]. AB - The aim of this study was to determine the rate of helminth infection in rudd, (Scardinus erythrophthalmus L. 1758) between spring and autumn seasons. A total of 87 rudd fish were investigated. Of these, 66 were infected with four species [Dactylogyrus difformis (Monogenea), Diplostomulum spathaceum metacerceria with Asymplodora markewitschi (Digenea), and Hysterothylacium sp. (Nematoda)]. During the study, it was noted that 46 out of 87 hosts were infected with D. difformis . The level of infection for this species was found to be 93% in the spring but it was 13% in the autumn. For D. spathaceum which was the second dominant species, 365 para-sites were found on 71 of 87 hosts. The maximum infection level for this species was 95% in the autumn. A total of 55 A. markewitschi could be found on 3 fish only during the spring. During the study, the lowest infection rate of the four species was that of Hysterothylacium sp. of which only 8 specimens were found on 7 fish. PMID- 17124669 TI - A new species of Monocystis stein, 1848 (Protista: Apicomplexa: Eugregarinida) from the Indian earthworm, Amynthas Hawayanus Rosa, 1891 (Annelida: Oligochaeta). AB - As a part of an ongoing biodiversity survey of aseptate gregarine fauna of oligochaete hosts of West Bengal, an expedition was carried out in the Darjeeling district of West Bengal and most of the earthworms collected were found to be infested with a species of Monocystis Stein, 1848. The monocystid species was collected from the seminal vesicles of the earthworm and was identified as a new species, Monocystis amynthae sp. nov. The gamont of the new species is characterized by having an elongated body with broad anterior end, separated from the narrow posterior end by a prominent constriction measuring 49.0-77.0 (66.0+/ 1.3) microm x 32.0-41.0 (37.0+/-2.8) microm. The gametocysts are oval-shaped, measuring 40.0-65.0 (58.0+/-2.1) microm. The oocysts are navicular, measuring 8.0 12.0 (10.5+/-1.1) microm x 4.0-6.0 (5.5+/-1.1) microm. PMID- 17124670 TI - Clinical, physiological and serological observations of a human following a venomous bite by Macrovipera lebetina lebetina (Reptilia: Serpentes). AB - A researcher, a 40 year-old male, was bitten on his finger accidentally by a 75 cm long, adult male blunt-nosed viper, Macrovipera lebetina lebetina (Linnaeus) that had been collected from northern Cyprus during laboratory work in the Zoology Section, Biology Department of the Ege University Faculty of Science. The clinical signs, physiological and biochemical changes soon after the venomous bite by the blunt-nosed viper as well as the electrophoretic separation of plasma proteins of the patient have been presented here. Serious important symptoms (edema, hypotension shock, hemorrhage, tissue necrosis, and melanoderma) developed and major quantitative differences in the fractions of albumin, globulin and albumin/globulin ratios were detected. On the other hand, the measurements carried out in blood smears and biochemical results indicated the decrease in RBC, MCV, MHV and a decrease in the diameter of red blood cells, as well as an increase in WBC. PMID- 17124671 TI - Age-dependent variations in the venom proteins of Vipera xanthina (Gray, 1849) (Ophidia: Viperidae). AB - In this study, polyacrylamide disc gel electrophoresis and densitometry analysis methods were used to analyze venom extracts of Vipera xanthina specimens of different lengths (35, 47 and 88 cm) collected from the same locality. The electropherograms of the venom protein samples showed age-dependent qualitative and quantitative variations. PMID- 17124672 TI - [Evaluation of the results of IgG avidity testing of Toxoplasma gondii in pregnant women.]. AB - Toxoplasmosis is a parasitic disease caused by a protozoon, Toxoplasma gondii. Infection acquired during pregnancy may cause intrauterine damage and sequelae in the newborn. In this study, IgG-avidity values were investigated in 695 pregnant women who were anti-T. gondii IgG antibody positive and the results obtained were evaluated. It has been found that the IgG of 492 women (70.8%) had high avidity, of 33 (4.7%), low avidity and that of 170 (24.5%) was in an equivocal range. Our results indicated that the IgG-avidity test must be carried out in pregnant women positive for anti-T. gondii IgG even if they are IgM antibody negative. PMID- 17124673 TI - [Investigation of Cryptosporidium spp. oocysts in tap water, well water, sewage water and sea water in Mersin, Turkey.]. AB - Cryptosporidium spp. are protozoa which only live in a host cell and may cause an infection that may result in the death of people with immune deficiency. It is known that Cryptosporidium parvum and Cryptosporidium hominis infections may be spread by contaminated well and tap waters. The facts that there is no certain and reliable cure and that the organisms may be found asymptomatically in the healthy people increases the importance of cryptosporidiosis. Our study has been performed in the city of Mersin and surrounding areas. A total of 100 samples of water were taken from taps (44 samples), wells (2 samples), the sea (35 samples) and sewage (19 samples) to investigate the presence of Cryptosporidium oocysts. Cryptosporidium oocysts have been detected in 5 samples of tap water, one sample of well water, one sample of sea water and 4 samples of sewage water. PMID- 17124674 TI - [Clinical symptoms in cases caused by entamoeba coli and blastocystis hominis.]. AB - E. coli and B. hominis are usually accepted as members of normal intestinal flora during stool examinations, but in recent years there has been controversy as to whether they may be pathogen protozoa. In this study, 92 individuals who were found to have E. coli (58/92) and B. hominis (34/92) in their stools were included in a study of clinical symptoms. No other parasitological or bacteriological agents were found in the stools of these persons. The percentages of intestinal symptoms were found to be 67.2% and 79.4% for E. coli and B. hominis, respectively. As a result of these findings we concluded that intestinal symptoms may be seen frequently if E. coli and B. hominis are present. In conclusion, E. coli and B. hominis may be considered to be pathogens, especially when no other agents are present. PMID- 17124675 TI - [Last updates on leishmania vaccine studies: when are we going to be vaccinated?]. AB - Approximately 350 million people in 8 countries are estimated to be under the threat of leishmaniasis. Most of the drugs used for the treatment of leishmaniasis are toxic and have many side effects. At present there is no vaccine against leishmaniasis. Vaccine development for parasitic infections is more difficult than for viruses and bacteria because of the antigenic complexity and parasite-host interactions. Anti-leishmanial vaccine candidates are reviewed in this paper. PMID- 17124676 TI - [A study of sister chromatid exchange (SCE) in patients with liver cystic hydatid disease who have been treated with albendazole.]. AB - A total of 21 patients including 13 females and 8 males who presented at the Microbiology and General Surgery Departments of the Ataturk Research and Training Hospital with complaints of liver cystic hydatid disease have been included in this study. Albendazole was administered to these cystic hydatid patients before and after surgery. The aim of this study was to investigate the probable genotoxic effects of albendazole. SCE testing was applied to the blood samples taken from patients after the treatment and the mutagenic effect of albendazole was investigated by comparison with the control group. Student t- test was used for the statistical analysis of the results. PMID- 17124677 TI - [Hydatid cyst prevalence in the region of Kars.]. AB - It has been reported that hydatid cysts are more common in societies that breed animals. Although there have been epidemiologic studies in various regions of our country, there has been no previous study related to this issue in the province of Kars where stockbreeding is quite common. In this study in order to determine the seroprevalence in the city of Kars and villages of the province of Kars, 511 serums were obtained and the specimens were analyzed by the indirect hemagglutination technique (IHAT) and indirect immunofluorescent technique (IFAT). The seropositive rate of 34.6% (177) found in this study, leads to the conclusion that hydatid cysts may be an important health problem in the region, and it was concluded that it would be suitable to carry out more comprehensive studies in the region. PMID- 17124678 TI - [Cystic echinococcosis in Kayseri during the last six years.]. AB - Cystic echinococcosis (CE) caused by the larval form of Echinococcus granulosus is a major public health problem especially in sheep-raising regions of the world. In the present study, CE cases detected during 1994-1998 by various hospitals and health directorships were investigated retrospectively. Our results show that there were a total of 699 cases of CE including 330 (47.2%) males and 369 (52.8%) females. Finally, it was found that the 699 patients had been hospitalized for a total of 9.246 days. PMID- 17124679 TI - [Analysis of anti-echinococcus antibodies in garbage men in Malatya.]. AB - It has been reported that some occupation groups constitute a higher risk group with regard to hydatid cyst disease. Municipal garbage men also are under this risk especially in those places where untamed dogs are abundant. The aim of this study was to analyze the anti-Echinococcus antibodies in garbage men in Malatya who are regarded as high risk-group with regard to hydatid cysts. Two hundred and forty serum specimens were collected and were later analyzed with the manual indirect hemagglutination technique (IHAT) and indirect immunofluorescent technique (IFAT). Of these sera 17 (7.08%) were found to be seropositive. Garbage men who were found to be seropositive were examined for intestinal parasites using the cellophane type technique, native Lugol and sedimentation methods. It was found that the group members giving seropositive results were free of parasites such as Taenia saginata and Hymenolepis nana which may lead to cross reaction with Echinococcus granulosus. The garbage men were informed about the results of the examination and were oriented for further examinations. The seropositive rate of 7.08% found in this study is supports the view that the garbage men should be regarded as a risk group. In order to inform all garbage men about means of protection, training seminar were organized. PMID- 17124680 TI - [Prevalence of hydatid cysts in cattle in the province of Kirikkale.]. AB - In present study, organs of cattle slaughtered in Kirikkale Province were investigated for cystic echinococcosis during a one year period. During this period, 847 cattle were examined and 120 of them were found to be infected with cystic echinococcosis. Hydatid cysts were located in the lung in 49.16% of the infected animals, in the liver in 16.68%, and in both the liver and lung in 34.16%. It was found that the number of hydatid cysts detected in 45 infected organs selected randomly (23 livers and 22 lungs) varied from 1-34 (average of 8.13). Fertile cysts were found in 3 (6.60%) organs (two lungs and one liver) during the entire study period. Sterile cysts were detected in 42 organs. Of the sterile cysts examined, 84.5% contained underdeveloped yellow germinal membranes and were filled with water. Of the remaining sterile cysts, 11.10% were caseous and 4.40%, calcified. Multivesicular cysts were found in 4.40% of organs examined. In this study, the prevalence of hydatid cysts was determined to be 14.16% in cattle in the Kirikkale Province. Since the fertility was low in the cysts examined, cattle play a limited role in Echinococcus epidemiology in this region. PMID- 17124681 TI - Toxocariasis seroprevalence in patients with rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease for which immunogenetic susceptibility factors have been defined. The aim of this study was to investigate seroprevalence of Toxocara canis (T. canis) in patients with RA. Forty-five patients with RA who were all diagnosed according to the ACR criteria and 48 health controls were included in this study. The mean age of patients with RA and of controls was 50+/-14 and 49+/-13, respectively. In both groups T. canis IgG antibodies were investigated with ELISA. Toxocara canis seroprevalence in patients with RA (35.6%) was significantly higher than that in the control group (8.3%) (p=0.001). There was no significant difference between patients with RA and control group in terms of behavioral factors (except in frequency of hand washing), demographic and socio-economic factors, while there was a difference in terms of owning dogs and cats or in the duration of ownership. When the patients with RA were divided into seropositive and seronegative, there were no significant differences between both groups in terms of demographic factors, socio-economic factors, behavioral factors, owning dogs and cats or duration of ownership and active or remission state of the illness, duration of disease and disability level in patients with RA (p > 0.05). In view of the present results, it is suggested that Toxocara infection was acquired before RA had developed and that high seroprevalence may result from exposure to pets and soil. It seems that this subject requires further investigations with larger numbers of samples in order to fully evaluate the causes of high seropositivity. PMID- 17124682 TI - [Distribution of intestinal parasites in children aged between 0 and 14 presenting at the Laboratory of Parasitology of the Mustafa Kemal University Medical School.]. AB - The aim of this study was to determine the parasite frequency in children from 0 14 years old who had presented at the parasitology laboratory of the Mustafa Kemal University Medicine School complaining of gastrointestinal and anal pruritus. During the laboratory investigation, 561 fecal specimens and 534 cellophane tapes obtained from the patients were examined. After the microscope examination of 561 feces samples, it was found that 104 of these samples (from 50 boys and 54 girls) contained one or more intestinal parasites. The number and distribution of these parasites in the specimens was as follows: 24 (4.27%) Giardia intestinalis, 9 (1.64%) Entamoeba histolytica, 3 (0.53%) Taenia saginata, 4 (0.71%) Dicrocoelium dendriticum, 51 (9.09%) Blastocystis hominis, 1 (0.17%) Enterobius vermicularis, 1 (0.17%) Entamoeba coli, 1 (0.17%) Entamoeba histolytica + Blastocystis hominis, 5 (0.89%) Giardia intestinalis + Blastocystis hominis, 1 (0.17%) Entamoeba coli + Blastocystis hominis, 1 (0.17%) Giardia intestinalis + Blastocystis hominis + Hymenolepis nana, 1 (0.17%) Blastocystis hominis + Enterobius vermicularis + Entamoeba coli. Out of the 534 cellophane tapes, 85 were found to be positive. PMID- 17124683 TI - [The distribution of intestinal parasites in students of the Mustafa Kemal University School of Health.]. AB - Intestinal parasites are an important national health problem in our country as in the rest of the world. In our study, the prevalence of intestinal parasites in female students (aged from 16-18 years) in the Mustafa Kemal University School of Health was investigated. Fecal samples and cellophane tape preparations were used for diagnosis. For this reason 142 fecal samples and 136 cellophane tape preparations were examined. One or more parasites were found in 65 (45.77%). fecal samples. The prevalence of parasites that found in fecal samples is as follows: Blastocystis hominis in 63 samples (96.92%) and Giardia intestinalis in 2 samples (3.08 %). Enterobius vermicularis was found in 9 (6.61%) out of 136 cellophane tape preparations. PMID- 17124684 TI - [Distribution of intestinal parasites among patients who presented at the Department of Parasitology of the Erciyes University Medical School.]. AB - In this study carried out from 2000-2004, a total of 34,883 stool samples were examined using native-Lugol and flotation/sedimentation methods and 9,879 cellophane tape preparations were examined directly. Intestinal parasites were found in 9,704 (27.8%) of the specimens. The parasites that were found and their prevalence is as follows: Blastocystis hominis, 6,723 (19.3%); Entamoeba coli, 1,007 (2.9%); Giardia intestinalis, 892 (2.6%); Entamoeba histolytica/E. dispar, 798 (2.3%); Endolimax nana, 486 (1.4%); Entamoeba hartmanni, 252 (0.7%); Enterobius vermicularis, 242 (0.7%); Iodamoeba butschlii, 109 (0.3%); Taenia saginata, 92 (0.3%); Chilomastix mesnili, 67 (0.2%); Ascaris lumbricoides, 55 (0.2%); and Hymenolepis nana, 40 (0.1%). PMID- 17124685 TI - Prevalence of intestinal parasitosis in children with gastrointestinal symptoms associated with socio-economic conditions in Manisa region. AB - Intestinal parasitosis is still an important public health problem. The aim of this study was to determine the prevalence of intestinal parasitosis in children with gastrointestinal symptoms, and to evaluate its association with socio economic and environmental factors. Stool samples of 3,216 children were examined by the saline-iodine method and trichrome staining. The cellophane tape method was also performed on 2,160 children. According to the educational levels and the economic status of families, the patients were classified as coming from underdeveloped, developing and developed areas. In 770 (23.9%) of 3,216 stool samples, various parasites were detected by the saline-iodine method and trichrome staining. The most common parasite was Giardia intestinalis (40.1%), followed by Entamoeba coli (10.2%). Enterobius vermicularis eggs were detected by the cellophane tape method in 221 (10.3%) out of 2,160 patients. The positive cases were evaluated according to the socio-economic and the environmental criteria; and most of them were found to have come from underdeveloped and developing areas. Health care and governmental officers should cooperate in order to improve the living conditions, and also people should be informed about the signs, symptoms and prevention methods of the parasitic diseases. PMID- 17124686 TI - [Distribution of intestinal parasites in children from the 23 Nisan Primary School in Hakkari.]. AB - This study was performed on children ranging from 6-14 years old coming from 23 Nisan Primary School in Hakkari. In this study, a total of 114 stool samples from 60 male students and 54 female students were examined for intestinal parasites. Stool containers were given to the students. After the stool containers were collected, they were taken to the Faculty of Medicine of the Yuzuncu Yil University and examined using native Lugol and flotation methods. One or more intestinal parasites were found in 66 (57.8%) out of a total of 114 students. Giardia intestinalis (28.9%), Blastocystis hominis (23.6%), Entamoeba coli (12.2%) and Ascaris lumbricoides (6.14%) were most prevalent parasites. PMID- 17124687 TI - [The remains of cattle slaughtered in the open fields provided the growth medium for the myiasis fly.]. AB - This study has been carried out to show that myiasis flies can grow on the carelessly discarded remains of slaughtered cattle. For this purpose, 11 larvae that were found on a piece of cattle remains that weighed 250 grams was put into 70% alcohol for identification; and the remaining 20 larvae were put into a pot containing sand on the bottom. The pot was covered with a piece of cloth in order to let them complete their growth. Later the pot was put outside so that the larvae could develop under natural conditions and daily growth of the larvae was determined. Pupas were produced by the larvae after 21 days and adult flies, after 28 days. With the help of identification keys, it was determined that out of the eleven larvae that were initially put into 70% alcohol, 5 larvae were third period Calliphora vicina and 3 larvae, second period. The remaining 3 larvae were third period Muscina stabulans. All of the 18 flies that developed from the 20 larvae in the covered pot outside were C. vicina. PMID- 17124688 TI - [Seroprevalence of hypodermatosis in cattle in the province of Sanliurfa (Turkey).]. AB - The aim of this study was to determine the seroprevalence of hypodermatosis in cattle in the province of Sanliurfa. For this purpose, sera samples were randomly collected from 300 cattle in the towns Sanliurfa, Suruc, Siverek, Viransehir and Akcakale. The sera were analyzed using a commercial ELISA kit. Out of the 300 cattle, 116 (38.6%) were found to be seropositive. Seropositivity rates ranged from 11.6-70.0 % according to age, race, gender and origins. In regard to race, significant difference (P < 0.001) was found between seropositivity ratios of pure breed and crossbreed animals. No significant difference was found between seropositivity ratios as a result of gender difference. Also, with regard to age and origins, significant differences (P < 0.01) were found between animals aged < 1 and aged>/=1 and in animals from Akcakale as well as Viransehir and other regions (P < 0.001). As a result, high seropositivity for hypodermatosis in cattle of the Sanliurfa Province has been detected. PMID- 17124689 TI - [Entomopoxviruses and biological control.]. AB - Poxviridae are divided into two subfamilies: the Chordopoxvirinae (poxviruses of vertebrates) and the Entomopoxvirinae (insect poxviruses). Entomopoxviruses (EPVs) are large (300-400 nm) oval shaped viruses. The genome of EPVs is large, with covalent ends and is a linear double-stranded DNA (200-240 kbp) molecule. The Entomopoxvirinae comprises three genera based on host insect and virion morphology. Genus A viruses infect coleopterans, genus B viruses infect lepidopterans and orthopterans, and genus C viruses infect dipterans. The Melolontha melolontha entomopoxvirus (MmEPV) was the first poxvirus to be described as being associated with an insect disease. Then, several entomopoxviruses (EPVs) have been found to infect Coleoptera, Lepidoptera, Diptera, Hymenoptera, and Orthoptera. Entomopoxviruses are very similar to orthopoxvirus and molluscipoxvirus that cause dermal lesions and pox diseases in humans. Therefore, these viruses have great importance in understanding their replication mechanism as well as in the use as a gene expression vector and as a pest control agent. In this review article, we present information about entomopoxviruses on which important studies have been done recently. PMID- 17124690 TI - [The importance of antinuclear (ABA) and anti-double stranded dna (anti-dsDNA) antibodies in the diagnosis of connective tissue diseases.]. AB - Connective tissue diseases (CTD) are characterized by the presence of autoantibodies against several tissues. These autoantibodies occur against cell membrane, cell receptors, plasma proteins, and cytoplasmic and nuclear components. In laboratories, anti-nuclear antibodies (ANA) and anti-double stranded DNA (anti-dsDNA) antibodies are widely used in diagnosis of CTD. The aim of this study was to investigate the presence and accompaniment of ANA and anti dsDNA antibodies in diagnosis of several CTD and also to study the prevalence of ANA and anti-dsDNA in a group of 88 patients with various types of CTD. ANA were detected by immunofluorescence (IFA) using HEp-2 cells (Zeus Scientific, Inc. USA) and anti-dsDNA antibodies using Crithidia luciliae (BioSystems, Spain) as substrates in immunofluorescence. ANA Western Blot (WB) Immunoassay (ImmuBlot, International Immuno-Diagnostics, USA) was also used along with the tests referred to previously. ANA was found in the sera of 84 (96.5%) patients while anti-dsDNA was detected in 7 (7.95%). Moreover different fluorescence patterns were also evaluated with ANA IFA in accordance with anti-dsDNA results. Mixed patterns in three and a homogeneous pattern in four anti-dsDNA positive patients' sera were determined on HEp-2 cell line by IFA. Seven sera which were ANA and anti-dsDNA positive with IFA were also found to be positive with WB and their ANA patterns with the specific ANA WB bands were also evaluated. It was observed that IFA results were in concordance with WB results. Our data indicated that the above findings should be controlled and evaluated with a more advanced method such as western blotting technique in order to confirm the presence of specific antibodies along with clinical outcome of the patients. As a result we think that ANA WB method is an appropriate technique in diagnosis of CTD as anti-dsDNA and ANA bands can be evaluated together with this method. PMID- 17124691 TI - [Seasonal distributions and effects of parasites in pike (esox lucius l., 1758) inhabiting the isikli dam lake (Denizli).]. AB - The aim of this study carried out from December 2000-November 2001 was to determine the endoparasites of pike (Esox lucius L., 1758) inhabiting the Isikli Dam Lake. A total of 160 samples were caught and investigated parasitologically. Bathybothrium rectangulum (Cestoda), Raphidascaris acus, Camallanus truncatus (Nematoda) and Neoechinorhynchus rutili (Acanthocephala) were detected as a result of this study. R. acus was the most prevalent species. The highest seasonal infection in the pike samples was determined to be 84.2% in the spring. The rates of infection are 40.4% in male pikes and 65.1% in females. According to age groups, the highest infection ratio has been determined to be 85.7% in pikes that were one year old. Investigations on samples caught during the same month and of the same age and gender showed that infected pikes were 2.5% shorter and 7.6% lighter than non-infected pikes. Among the parasites determined in this study, Bathybothrium rectangulum was detected for the first time in Turkish pike. PMID- 17124692 TI - Pseudomazzia macrolabiata n.gen.,n.sp. (Nematoda: Spiruridae: Mazziinae) from the fish Pomadasys olivaceus of Karachi coast. AB - A nematode Pseudomazzia macrolabiata of a new genus and a new species is described here from the intestine of the fish Pomadasys olivaceus of Karachi coast. The new genus is characterized by having an enlarged head region with three large prominent lips protruding externally from the head, a prominent buccal cavity and with a proboscis. The esophagus is not clearly divided into muscular and glandular parts and the intestine is simple. The male has very unequal spicules, dissimilar in shape. The caudal alae are prominent supported by several pedunculate papillae including 5 pre anal pairs and two pairs including one pro-anal and one post-anal. The female vulva is near the middle of the body. The tail is elongate, bluntly pointed and is provided with a terminal forked spike. The eggs are thin shelled and oval in shape. PMID- 17124693 TI - Use of a confidence scale in reporting postmortem fetal magnetic resonance imaging. AB - OBJECTIVES: Postmortem magnetic resonance imaging (MRI) may be an alternative to conventional autopsy. However, it is unclear how confident radiologists are in reporting such studies. We sought to determine the confidence with which radiologists report on various fetal organs by developing a scale to express their confidence of normality and abnormality, and to place this in the context of a pathological diagnosis of whether the organ was in fact normal or abnormal. METHODS: Thirty fetuses, aged 16-39 gestational weeks and weighing 61-3270 g, underwent postmortem MRI prior to conventional autopsy. MRI studies were reported by two radiologists with access to the clinical and sonographic history: a neuroradiologist, reporting head and neck, and a pediatric radiologist, reporting thorax, abdomen and pelvis. Radiologists used a scale (0 = definitely abnormal, 100 = definitely normal, 50 = unable to comment) to indicate their confidence of anatomical structures being normal or abnormal, using a checklist. Conventional autopsies were performed by pediatric pathologists blinded to the MRI findings, and these were considered the reference standard. RESULTS: Most normal fetal organs had high scores on postmortem MRI, with median confidence scores above 80. However, the atrioventricular valves, duodenum, bowel rotation and pancreas proved more difficult to assess, with median scores of 50, 60, 60 and 62.5, respectively. Abnormal cardiac atria and ventricles, kidneys, cerebral hemispheres and corpus callosum were always detected with high or moderate degrees of confidence (median scores of 2.5, 5, 0, 0 and 30 respectively). However, in two cases with abnormal cardiac outflow tracts, both cases scored 50. Kappa values, assessing agreement between MRI diagnoses of abnormality and autopsy, were high for the brain (0.83), moderate for the lungs (0.56) and fair for the heart (0.33). CONCLUSIONS: This scoring system represents an attempt to define the confidence of radiologists to report varying degrees of normality and abnormality following z ex-utero fetal MRI. While most fetal anatomy is clearly visualized on postmortem MRI, radiologists may lack confidence reporting such studies and there are particular problems with assessment of some cardiac and gastrointestinal structures, both normal and abnormal. PMID- 17124694 TI - Appeals by the elderly against compulsory detention under the Mental Health Act 1983. PMID- 17124695 TI - Factors associated with an unusual increase in the elderly suicide rate in Taiwan. PMID- 17124696 TI - An audit of service utilization by graduates attending an old age psychiatry service. PMID- 17124697 TI - Patients and relatives desire their physician to give a judgement about driving abilities: a survey by questionnaire on a Dutch memory clinic. PMID- 17124698 TI - An approach to joint analysis of longitudinal measurements and competing risks failure time data. AB - Joint analysis of longitudinal measurements and survival data has received much attention in recent years. However, previous work has primarily focused on a single failure type for the event time. In this paper we consider joint modelling of repeated measurements and competing risks failure time data to allow for more than one distinct failure type in the survival endpoint which occurs frequently in clinical trials. Our model uses latent random variables and common covariates to link together the sub-models for the longitudinal measurements and competing risks failure time data, respectively. An EM-based algorithm is derived to obtain the parameter estimates, and a profile likelihood method is proposed to estimate their standard errors. Our method enables one to make joint inference on multiple outcomes which is often necessary in analyses of clinical trials. Furthermore, joint analysis has several advantages compared with separate analysis of either the longitudinal data or competing risks survival data. By modelling the event time, the analysis of longitudinal measurements is adjusted to allow for non ignorable missing data due to informative dropout, which cannot be appropriately handled by the standard linear mixed effects models alone. In addition, the joint model utilizes information from both outcomes, and could be substantially more efficient than the separate analysis of the competing risk survival data as shown in our simulation study. The performance of our method is evaluated and compared with separate analyses using both simulated data and a clinical trial for the scleroderma lung disease. PMID- 17124699 TI - A residuals-based transition model for longitudinal analysis with estimation in the presence of missing data. AB - We propose a transition model for analysing data from complex longitudinal studies. Because missing values are practically unavoidable in large longitudinal studies, we also present a two-stage imputation method for handling general patterns of missing values on both the outcome and the covariates by combining multiple imputation with stochastic regression imputation. Our model is a time varying auto-regression on the past innovations (residuals), and it can be used in cases where general dynamics must be taken into account, and where the model selection is important. The entire estimation process was carried out using available procedures in statistical packages such as SAS and S-PLUS. To illustrate the viability of the proposed model and the two-stage imputation method, we analyse data collected in an epidemiological study that focused on various factors relating to childhood growth. Finally, we present a simulation study to investigate the behaviour of our two-stage imputation procedure. PMID- 17124701 TI - Local atomic order and infrared spectra of biogenic calcite. PMID- 17124702 TI - Comparison of thermosensitive properties of poly(amidoamine) dendrimers with peripheral N-isopropylamide groups and linear polymers with the same groups. PMID- 17124703 TI - The fight against new types of influenza virus. AB - In 1997, during an outbreak in chickens in Hong Kong the avian H5N1 influenza virus crossed the species barrier and infected 18 people, of which 6 cases were fatal. The virus also infected wild birds and continued to circulate and mutate in geese and ducks in southeastern China. Since this occurrence, new antigenic variants that are highly pathogenic for humans as well as wild, domestic, and exotic waterfowl continue to appear in Hong Kong. This virus is spreading across Asia, and is encroaching upon Europe and other continents. Wild birds are now considered as the main reservoir of H5N1 virus. Humans become infected with this H5N1 virus usually via close contact with infected birds or a highly contaminated environment. The very low transmissibility of this virus prevented further person to-person dissemination in spite of the complete absence of immunity in the human population to H5N1 viruses. Viruses of the H5N1 subtype are characterized by an exceptionally high pathogenicity for humans. The cause of the viral virulence is not known so far; however, several virulence factors are considered. The unprecedented capability of H5N1 viruses to kill humans intensifies the concern about its pandemic potential with catastrophic consequences. The effectiveness of existing antivirals as well as vaccines for humans and birds are reviewed. PMID- 17124704 TI - Molecular characterization of rpoB gene mutations in rifampicine-resistant Mycobacterium tuberculosis isolates from tuberculosis patients in Belarus. AB - The aim of this study was to investigate the frequency, location and type of rpoB mutations in Mycobacterium tuberculosis isolated from patients in Belarus. Tuberculosis cases are increasing every year in Belarus. Moreover, resistance to anti-tuberculosis drugs, especially to rifampicine, has increased. In this study, 44 rifampicine-resistance M. tuberculosis clinical isolates (including multidrug resistant isolates) were subjected to DNA sequencing analysis of the hypervariable region (hot-spot) of the rpoB gene originating from different geographical regions in Belarus. Sixteen different types of mutations were identified. The most common point mutations were in codons 510 (47.7%), 526 (45.5%), 523 (40.86%) and 531 (29.5%). Eleven isolates (27.7%) carried one mutation and 23 (52%), 7 (16%), 3 (7%) of isolates carried 2, 3 and 4 mutations, respectively. A characteristic, prominent finding of this study was high frequency of multiple mutations in different codons of the rpoB gene (27.7%) and also the detection of unusual types of mutations in the 510 codon, comprising CAG mutations (deletion or changing, to CTG, CAC or CAT). In our study, the change TTG in codon 531 was found in 92% of isolates and the change TGC in 8% of isolates. A TAC change in codon 526 was not found, but the GAC change was found in all isolates. Isolates of M. tuberculosis isolated in Belarus were characterized by the wide spectrum of the important mutations and might belong to the epidemic widespread clones. PMID- 17124705 TI - Subtractive hybridization magnetic bead capture: a new technique for the recovery of full-length ORFs from the metagenome. AB - A new method for the recovery of full-length open reading frames from metagenomic nucleic acid samples is reported. This technique, based on subtractive hybridization magnetic bead capture technology, has the potential to access multiple gene variants from a single amplification reaction. It is now widely accepted that classical microbiological methods provide only limited access to the true microbial biodiversity (less than 1%). The desire to access a higher proportion of the metagenome has led to the development of efficient environmental nucleic acid extraction technologies and to a range of sequence dependent and sequence-independent gene discovery techniques. These methods avoid many of the limitations of culture-dependent gene targeting. PMID- 17124706 TI - Consumer acceptance of ingenic foods. AB - Recent advances in plant molecular biology offer a means of reaping the benefits of biotechnology, while potentially assuaging consumer concerns by re-inserting native DNA back into plants. Results are presented from nationwide surveys in the U.S. and France, indicating that more consumers would accept ingenic plants than transgenic plants, with twice as many U.S. than French consumers considering food produced through biotechnology eatable. PMID- 17124707 TI - Meeting report: protein design and evolution for biocatalysis August 30 - September 1, 2006, Greifswald, Germany. PMID- 17124708 TI - Electrokinetic transport of charged solutes in micro- and nanochannels: the influence of transverse electromigration. AB - The accurate prediction of electrokinetic migration velocity and dispersion is crucial to separating electrophoretically charged solutes in micro- or nanochannels. In this paper, we investigate numerically the influence of transverse electromigration (TEM) on the solute electrokinetic transport in a series of micro- and nanochannels. The TEM, often ignored in previous studies, is demonstrated to significantly affect the solute migration velocity in nanochannels and the electrokinetic dispersion in microchannels. This is because the TEM can force either positively charged solutes into or negatively charged solutes out of the electrical double layer that forms adjacent to the negatively charged channel wall and contains the velocity gradients. Analytical solutions are also derived for characterizing the electrokinetic transport of charged solutes in nanochannels, which has been validated to be in good agreement with the numerical simulation. Moreover, we demonstrate that the proposed analytical formula for the solute migration velocity actually applies to channels of any size. PMID- 17124709 TI - Viability study of HL60 cells in contact with commonly used microchip materials. AB - This paper presents a study in which different commonly used microchip materials (silicon oxide, borosilicate glass, and PDMS) were analyzed for their effect on human promyelocytic leukemic (HL60) cells. Copper-coated silicon was analyzed for its toxicity and therefore served as a positive control. With quantitative PCR, the expression of the proliferation marker Cyclin D1 and the apoptosis marker tissue transglutaminase were measured. Flow cytometry was used to analyze the distribution through the different phases of the cell cycle (propidium iodide, PI) and the apoptotic cascade (Annexin V in combination with PI). All microchip materials, with the exception of Cu, appeared to be suitable for HL60 cells, showing a ratio apoptosis/proliferation (R(ap)) comparable to materials used in conventional cell culture (polystyrene). These results were confirmed with cell cycle analysis and apoptosis studies. Precoating the microchip material surfaces with serum favor the proliferation, as demonstrated by a lower R(ap) as compared to uncoated surfaces. The Cu-coated surface appeared to be toxic for HL60 cells, showing over 90% decreased viability within 24 h. From these results, it can be concluded that the chosen protocol is suitable for selection of the cell culture material, and that the most commonly used microchip materials are compatible with HL60 culturing. PMID- 17124710 TI - Towards a portable microchip system with integrated thermal control and polymer waveguides for real-time PCR. AB - A novel real-time PCR microchip platform with integrated thermal system and polymer waveguides has been developed. The integrated polymer optical system for real-time monitoring of PCR was fabricated in the same SU-8 layer as the PCR chamber, without additional masking steps. Two suitable DNA binding dyes, SYTOX Orange and TO-PRO-3, were selected and tested for the real-time PCR processes. As a model, cadF gene of Campylobacter jejuni has been amplified on the microchip. Using the integrated optical system of the real-time PCR microchip, the measured cycle threshold values of the real-time PCR performed with a dilution series of C. jejuni DNA template (2 to 200 pg/microL) could be quantitatively detected and compared with a conventional post-PCR analysis (DNA gel electrophoresis). The presented approach provided reliable real-time quantitative information of the PCR amplification of the targeted gene. With the integrated optical system, the reaction dynamics at any location inside the micro reaction chamber can easily be monitored. PMID- 17124711 TI - Activation of the disulfide bond and chalcogen-chalcogen interactions: an experimental (FTICR) and computational study. AB - Dimethyldisulfide (I) is the simplest model of the biologically relevant family of disubstituted disulfides. The experimental study of its gas-phase protonation has provided, we believe for the first time, a precise value of its gas-phase basicity. This value agrees within 1 kJ mol-1 with the results of G3 calculations. Also obtained for the first time was the reaction rate constant for the bimolecular reaction between I and its protonated form, IH+, to yield methanethiol and a dimethyldithiosulfonium ion. This constant is of the order of magnitude of the collision limit. A computational mechanistic study based on the energetic profile of the reaction, completed with Fukui's and Bader's treatments of the reactants and transition states fully rationalizes the regioselectivity of the reaction as well as the existence of a shallow, flat Gibbs energy surface for the reaction. The mechanistic relevance of the chalcogen-chalcogen interaction and the C--H...S bonds has been demonstrated. PMID- 17124712 TI - Reactions of hydrogen atoms with met-enkephalin and related peptides. AB - The reactions of hydrogen atoms with enkephalins and related peptides have been investigated by radiolytic methods in aqueous solutions and lipid vesicle suspensions. Pulse radiolysis experiments indicate that methionine residue (Met) is the main target. In Met-enkephalin (Tyr-Gly-Gly-Phe-Met) the attack of the hydrogen atom occurs to about 50 % on Met with formation of methanethiyl radical. The remaining percentage is divided roughly evenly between Tyr and Phe. With Leu enkephalin (Tyr-Gly-Gly-Phe-Leu) the site of attack is limited to Tyr and Phe. Using a peptide-liposome (that is, 1-palmitoyl-2-oleoyl phosphatidylcholine vesicles) model, the cis-trans isomerization of phospholipids could be detected due to the catalytic action of thiyl radicals. The radiation chemical yields of the H(.) and, consequently of CH(3)S(.) radical, was modulated by the experimental conditions and the nature of peptide. Large amounts of trans lipids observed in phosphate buffer vesicle suspensions indicated the efficient role of double-bond isomerization as marker of Met-containing peptide damage. PMID- 17124713 TI - Nanocomposite and nanoporous polyaniline conducting polymers exhibit enhanced catalysis of nitrite reduction. AB - Nanostructured polyaniline (PANI) conducting polymer films were prepared on electrochemically pretreated glassy carbon electrodes, which were previously modified with multilayers of polystyrene (PS) nanoparticles with a diameter of 100 nm. PANI was electropolymerised and grown through the interstitial spaces between the PS nanoparticles, which formed a nanocomposite film of PANI and PS nanoparticles on the electrode surface. Furthermore, a nanoporous PANI film was fabricated through the removal of the PS nanoparticles by dissolution in toluene. As a result of their nanostructure, both of the PANI films (before and after removal of the PS nanoparticles) exhibited enhanced electrocatalytic behaviour towards the reduction of nitrite relative to bulk-PANI films; however, partial collapse or shrinkage may have occurred with the removal of the nanoparticles and could have resulted in a less enhanced response. Under optimised conditions, the nanocomposite-film-modified electrode exhibited a fast response time of 5 s and a linear range from 5.0 x 10(-7) to 1.4 x 10(-3) M for the detection of nitrite; the detection limit was 2.4 x 10(-7) M at a signal-to-noise ratio of 3. PMID- 17124714 TI - Total synthesis and evaluation of the actin-binding properties of microcarpalide and a focused library of analogues. AB - A comparative investigation shows that hydroxylated 10-membered lactones modeled around the fungal metabolites microcarpalide (1) and pinolidoxin (2) are endowed with selective actin-binding properties. Although less potent than the marine natural product latrunculin A, which represents the standard in the field, nonenolides of this type are significantly less toxic and accommodate substantial structural editing. Most notable is the fact that even an intramolecular transesterification with formation of a hydroxylated butanolide skeleton does not annihilate their microfilament disrupting capacity. This finding calls for a reinvestigation of the biological profile of other fungal metabolites that embody a similar motif. Microcarpalide (1) serving as the calibration point for this comparative study was prepared by total synthesis based on ring-closing metathesis (RCM) as the key step. The chosen route favorably compares to previous approaches to this target and provides further support for the notion that the (E,Z)-configuration of a medium-sized cycloalkene can be controlled by proper choice of the catalyst as previously outlined by our group. 9-epi-Microcarpalide 26 and furanone 27 as representative examples of the "natural productlike" compounds investigated herein have been characterized by crystal structure analysis. PMID- 17124715 TI - Computer-aided rational design of catalytic antibodies: The 1F7 case. PMID- 17124716 TI - Preservation of plant genetic resources in the biotechnology era. AB - Thousands of years ago humans began domesticating crops as a food source. Among the wild germplasm available, they selected those that were best adapted for cultivation and utilization. Although wild ancestors have continued to persist in regions where domestication took place, there is a permanent risk of loss of the genetic variability of cultivated plants and their wild relatives in response to changing environmental conditions and cultural practices. Recognizing this danger, plant ex situ genebank collections were created since the beginning of the last century. World-wide, more than 6 million accessions have been accumulated including the German ex situ genebank in Gatersleben, one of the four largest global collections, housing 150,000 accessions belonging to 890 genera and 3032 species. This review summarizes the ex situ plant genetic resources conservation behavior with a special emphasis on German activities. Strategies for maintenance and management of germplasm collections are reviewed, considering modern biotechnologies (in vitro and cryo preservation). General aspects on genetic diversity and integrity are discussed. PMID- 17124717 TI - An investigation of the effects of endometriosis on the proteome of human eutopic endometrium: a heterogeneous tissue with a complex disease. AB - The pathogenesis of endometriosis includes the proliferation of heterogeneous endometrial cells and their invasion into ectopic sites within the peritoneal cavity. This may be due to abnormalities of the eutopic endometrium itself, predisposing the cells to survive and implant ectopically. We investigated the applicability of 2-DE gels and peptide mass mapping to identify candidate endometrial proteins with a role in endometriosis. Despite the heterogeneous nature of endometrium, our results show that combining the analysis of 2-DE gels and peptide mass mapping yields consistent data. We identified dysregulated proteins in women with endometriosis which included: (i) molecular chaperones including heat shock protein 90 and annexin A2, (ii) proteins involved in cellular redox state, such as peroxiredoxin 2, (iii) proteins involved in protein and DNA formation/breakdown, including ribonucleoside-diphosphate reductase, prohibitin and prolyl 4-hydroxylase, and (iv) secreted proteins, such as apolipoprotein A1. These proteins have functions which suggest that they could play a role in the pathogenesis of endometriosis. This study demonstrated that 2 DE gel analysis and mass spectroscopic protein identification are suitable for the identification of proteins with candidate associations with endometriosis. These techniques should be used on a larger scale to identify endometriosis related proteins, thus improving the understanding of this complex disease. PMID- 17124718 TI - Establishing a liquid-phase IEF in combination with 2-DE for the analysis of Leishmania proteins. AB - The recent completion of genome sequencing projects for Leishmania major and near completion for two other species, L. infantum and L. braziliensis, has provided the needed genomic information for investigating the proteomes of Leishmania parasites. However, the design of effective 2-DE-based proteome mapping for complex protozoan parasites like Leishmania has proven to be severely compromised due to extensive overcrowding of spots especially in the acidic regions, coupled to a relatively low representation of basic proteins. In the present study, we optimized a liquid-phase IEF in combination with 2-DE for L. amazonensis promastigote as a way of reducing protein complexity and enhancing representation for low-abundance proteins on gels. Of 20 pH-based fractions eluted from Rotofor cells, 5 representative fractions selected from acidic, basic or neutral regions of the proteome and with adequate protein concentration were further analyzed by 2-DE using medium-range IPG strips. On this basis, we were able to generate high resolution 2-DE maps encompassing both the acidic and basic ends of the proteome with enhanced spot representation. PMID- 17124719 TI - Real time PCR profiling of 330 human micro-RNAs. AB - The small size of miRNAs makes profiling of all the 462 known human miRNAs difficult using single cell samples. Recently, we showed that judicious sequence partitioning between RT primers and second strand synthesis primers permitted multiplexed RT-PCR amplification of miRNA in very small samples to allow individual real time PCR quantification. Here, we show that zip coding the primers and TaqMan probes with sequences specific to each miRNA greatly improves reaction specificity, which permits the profiling of all miRNAs in a single multiplexed RT-PCR reaction. PMID- 17124720 TI - Treatment of new-onset type 1 diabetes with peptide DiaPep277 is safe and associated with preserved beta-cell function: extension of a randomized, double blind, phase II trial. AB - BACKGROUND: Treatment with DiaPep277, a peptide derived from HSP60, has been shown to preserve beta-cell function in non-obese diabetic mouse (NOD) mice and in a trial with newly diagnosed human patients with type 1 diabetes treated over a 10-month period. This article extends the clinical trial observations to a total of 20 months of treatment to determine the safety and the effects of repeated doses of DiaPep277 on endogenous insulin secretion, metabolic control, and exogenous insulin requirements. METHODS: Thirty-five male patients (aged 16 58) with a basal C-peptide greater than 0.1 nmol/L were assigned to periodic treatment with DiaPep277 (1 mg) or placebo for a 12-month treatment and 18-month observation protocol, later extended to an additional year of treatment. Stimulated C-peptide, HbA1c, and an exogenous insulin dose were the clinical endpoints. RESULTS: At 18 months, stimulated C-peptide concentrations had fallen in the placebo group (p = 0.0005) but were maintained in the DiaPep277 group. The need for exogenous insulin was higher in the placebo group than in the DiaPep277 group. Mean HbA1c concentrations were similar in both groups. After extension of the study, patients continuing treatment with DiaPep277 and those switched from placebo to DiaPep277 manifested a trend towards a greater preservation of beta cell function compared to patients maintained on or switched to placebo. The safety profile of DiaPep277 was similar between the treatment and placebo groups, and no drug-related adverse events occurred. CONCLUSIONS: Periodic treatment of subjects with DiaPep277 over 2 years was safe and associated preservation of endogenous insulin secretion up to 18 months was observed. PMID- 17124721 TI - Heat-shock protein peptide DiaPep277 treatment in children with newly diagnosed type 1 diabetes: a randomised, double-blind phase II study. AB - BACKGROUND: Type 1 diabetes mellitus (T1DM) is a T-cell-mediated autoimmune disease that leads to the destruction of insulin-producing beta cells. Treatment with DiaPep277, a peptide derived from heat-shock protein 60 (hsp60), has been found to slow the deterioration of beta-cell function after clinical onset of diabetes in NOD mice and human adults. Our aim was to evaluate the efficacy and safety of DiaPep277 treatment in attenuating beta-cell destruction in children with recent-onset T1DM. METHODS: A prospective, randomized, double-blind, phase II design was used. The sample included 30 children (19 males) aged 7-14 years who had been diagnosed with T1DM from 53 to 116 days previously, and had basal C peptide concentrations above 0.1 nmol/L. The children were randomized to receive subcutaneous injections of 1 mg DiaPep277 (15 patients) or 40 mg mannitol (placebo) at entry and at 1, 6, and 12 months. The duration of follow-up was 18 months. The groups were compared for stimulated C-peptide level, exogenous insulin dose, and HbA1c concentration. RESULTS: C-peptide levels similarly decreased over time in the DiaPep277- and placebo-treated patients. There was no significant difference in insulin dose or HbA1c concentration between the groups at any time point. No serious drug-related adverse effects were recorded throughout the study period. CONCLUSIONS: One-year treatment with DiaPep277 at a dosage of 1 mg is safe for use and well tolerated in children with recent-onset T1DM. However, it appears to have no beneficial effect in preserving beta-cell function or improving metabolic control. PMID- 17124722 TI - I've just been diagnosed with atherosclerosis. Is there anything I can do to unclog my arteries? PMID- 17124723 TI - Hospital charged with dumping homeless patient. PMID- 17124725 TI - Nothing to declare? PMID- 17124724 TI - Our great depression. PMID- 17124726 TI - A protein alternative to RNAi. AB - A general method of targeting tagged proteins for rapid degradation in the cell promises to provide a powerful alternative to RNA interference (RNAi) for studying the functions of proteins in living cells. PMID- 17124727 TI - Medicine meets religion in organ donation debate. PMID- 17124728 TI - Story of C's. AB - Chromosome conformation capture (3C) graduates to a tool for mapping interaction networks, thanks to a clever way of making molecular 'carbon copies'. PMID- 17124729 TI - Nicaragua eliminates last exception to strict anti-abortion law. PMID- 17124730 TI - The inside tag. AB - An uncharged CoA precursor that can enter the cell is used for covalent, site specific labeling of proteins inside living cells. PMID- 17124731 TI - Officials clash over mentally ill in Florida jails. PMID- 17124732 TI - Bacteria make the rotor go round. AB - The gliding bacteria Mycoplasma mobile yoked to an inorganic rotor will turn it along a circular track, generating a tiny biological hybrid motor. PMID- 17124733 TI - Sweetening up yeast glycoprotein synthesis. AB - An engineered yeast strain is capable of carrying out the full range of human protein glycosylation reactions. PMID- 17124734 TI - siRNAs meet their match. AB - A new study addresses how to design highly specific small interfering RNAs (siRNAs) capable of distinguishing between similar variants of the same gene. PMID- 17124735 TI - Rational design of a super core promoter that enhances gene expression. AB - Transcription is a critical component in the expression of genes. Here we describe the design and analysis of a potent core promoter, termed super core promoter 1 (SCP1), which directs high amounts of transcription by RNA polymerase II in metazoans. SCP1 contains four core promoter motifs-the TATA box, initiator (Inr), motif ten element (MTE) and downstream promoter element (DPE)-in a single promoter, and is distinctly stronger than the cytomegalovirus (CMV) IE1 and adenovirus major late (AdML) core promoters both in vitro and in vivo. Each of the four core promoter motifs is needed for full SCP1 activity. SCP1 is bound efficiently by TFIID and exhibits a high propensity to form productive transcription complexes. SCP1 and related super core promoters (SCPs) with multiple core promoter motifs will be useful for the biophysical analysis of TFIID binding to DNA, the biochemical investigation of the transcription process and the enhancement of gene expression in cells. PMID- 17124740 TI - Lousy mood. A link to blood glucose. PMID- 17124739 TI - National recommendations for donation after cardiocirculatory death in Canada: Donation after cardiocirculatory death in Canada. AB - These recommendations are the result of a national, multidisciplinary, year-long process to discuss whether and how to proceed with organ donation after cardiocirculatory death (DCD) in Canada. A national forum was held in February 2005 to discuss and develop recommendations on the principles, procedures and practice related to DCD, including ethical and legal considerations. At the forum's conclusion, a strong majority of participants supported proceeding with DCD programs in Canada. The forum also recognized the need to formulate and emphasize core values to guide the development of programs and protocols based on the medical, ethical and legal framework established at this meeting. Although end-of-life care should routinely include the opportunity to donate organs and tissues, the duty of care toward dying patients and their families remains the dominant priority of health care teams. The complexity and profound implications of death are recognized and should be respected, along with differing personal, ethnocultural and religious perspectives on death and donation. Decisions around withdrawal of life-sustaining therapies, management of the dying process and the determination of death by cardiocirculatory criteria should be separate from and independent of donation and transplant processes. The recommendations in this report are intended to guide individual programs, regional health authorities and jurisdictions in the development of DCD protocols. Programs will develop based on local leadership and advance planning that includes education and engagement of stakeholders, mechanisms to assure safety and quality and public information. We recommend that programs begin with controlled DCD within the intensive care unit where (after a consensual decision to withdraw life-sustaining therapy) death is anticipated, but has not yet occurred, and unhurried consent discussions can be held. Uncontrolled donation (where death has occurred after unanticipated cardiac arrest) should only be considered after a controlled DCD program is well established. Although we recommend that programs commence with kidney donation, regional transplant expertise may guide the inclusion of other organs. The impact of DCD, including pre-and post-mortem interventions, on donor family experiences, organ availability, graft function and recipient survival should be carefully documented and studied. PMID- 17124741 TI - Ties that bind. The pancreas piece of the diabetes puzzle. PMID- 17124742 TI - Atrial fibrillation. A little-known big problem. PMID- 17124743 TI - The season of giving. Great taste and good nutrition are the perfect gifts for everyone on your list. PMID- 17124744 TI - Pedicure confidential. PMID- 17124745 TI - Research profile. Laying groundwork. Foot complications and dialysis. Lawrence Lavery, DPM, MPH. PMID- 17124746 TI - Bah humbug! Tips for fighting the holiday blues. PMID- 17124747 TI - Ketones gone wild. Understanding ketoacidosis. PMID- 17124748 TI - Healthy eating. Be of good cheer. Holiday entertaining made easy. PMID- 17124750 TI - Drained. (Anemia and diabetes). PMID- 17124749 TI - Aging gracefully. How to live well with diabetes as you get older. PMID- 17124751 TI - Family ties. Diabetes during pregnancy carries a lifelong risk of type 2 for mother and child. PMID- 17124752 TI - Muffin madness. Bake up some healthy, tasty breakfast muffins. PMID- 17124753 TI - Research profile. Risky business. Heart disease and type 1 diabetes. Janice C. Zgibor, PhD. PMID- 17124754 TI - [Hepatitis induced by fibrates]. PMID- 17124755 TI - WHO position paper on Haemophilus influenzae type b conjugate vaccines. (Replaces WHO position paper on Hib vaccines previously published in the Weekly Epidemiological Record. PMID- 17124756 TI - The changing view of neurorehabilitation: a new era of optimism. PMID- 17124757 TI - The recognition of emotional expression in prosopagnosia: decoding whole and part faces by Stephan, Breen and Caine. PMID- 17124758 TI - Human embryonic stem cell research: an argument for national research review. AB - The US National Academy of Sciences (NAS) recently published voluntary guidelines for human embryonic stem (hES) cell research. The NAS guidelines propose two levels of oversight. AT the local level, research institutions are to create Embryonic Stem Cell Research Oversight (ESCRO) committees with a mandate to assess the scientific merit and ethical acceptability of hES cell research. At the national level, a new committee is to be created, not to review specific research proposals, but rather to periodically assess, and as needed revise, the NAS guidelines. In this article, we critically assess this proposal. In particular, we review the benefits and limitations of local research review. On this basis, we argue that local review is insufficient for hES cell research and that while there are obvious pragmatic and political reasons for the NAS to favor local research review, there are more compelling reasons for the NAS to have recommended national review of hES cell research proposals. PMID- 17124759 TI - Research misconduct and the scientific process: continuing quality improvement. AB - The response to research misconduct involves the attempt to regulate behavior through (a) creating and enforcing a rule and (b) ethics education. The roles of each must be shaped by considerations of the nature of scientific practice. Given the nature of science, the role of (a) must be limited in scope: both in the types of behavior it covers and in the level of intent that must be present for an allegation of misconduct to be proven. Since one important role of ethics education is to fill the gaps that regulatory rules leave open, it is this limitation in scope and its source in theoretical concerns that better reveals the type and kind of education needed. It is argued that much of the current ethics education falls short. Since the gaps left by the rule are largely due to theoretical concerns about the very nature of the scientific process and the nature of that process is constantly evolving, ethics education must focus more heavily on theory and must reach a wider audience. It is argued that ethics education can be more effective if it aims, in part, in creating a discipline specific, constantly evolving standard of care. PMID- 17124760 TI - Research misconduct policies of high impact biomedical journals. AB - Several national and international organizations have recommended policies regarding journal responsibilities about research misconduct in submitted or published manuscripts. A search of Web sites of the fifty highest impact journals in a cluster of biomedical fields and a limited survey of their editors shows that few journals have formally adopted standards for dealing with questions of research misconduct. Publicly available policies may have a deterrent effect and can prevent arbitrariness in handling cases. PMID- 17124761 TI - Deception in psychology: moral costs and benefits of unsought self-knowledge. AB - Is it ethically permissible to use deception in psychological experiments? We argue that, provided some requirements are satisfied, it is possible to use deceptive methods without producing significant harm to research participants and without any significant violation of their autonomy. We also argue that methodological deception is at least at the moment the only effective means by which one can acquire morally significant information about certain behavioral tendencies. Individuals in general, and research participants in particular, gain self-knowledge which can help them improve their autonomous decision-making. The community gains collective self-knowledge that, once shared, can play a role in shaping education, informing policies and in general creating a more efficient and just society. PMID- 17124762 TI - Two facets of peer review and the proper role of study sections. AB - The current National Institutes of Health study section system is under increasing criticism due to tight budgets and decreased levels of perceived competence. There is also an overemphasis on written critiques from the study section by unsuccessful applicants. It is argued that this arises from confusion between two different purposes of peer review. A system of universal participation in peer review by senior funded investigators is proposed to ameliorate these problems. PMID- 17124763 TI - Theoretical issues in the regulation of genetically engineered organisms: the case of deliberate release. AB - At the cutting edge of research in biotechnology is the production of special genetically engineered organisms for use in an open, agricultural environment. We argue that the standard approaches to risk assessment and management employed by regulatory agencies are inadequate to the task of properly assessing the advisability of such experiments, as well as the even more significant prospect of widespread commercial use of genetically engineered organisms in agriculture. Given the current lack of sufficient information with which to make competent assessments of the risks involved and the attempts to gain such information in a timely fashion, we conclude that a compelling case for deliberate release programs has not been made. PMID- 17124764 TI - Theoretical issues in the regulation of genetically engineered organisms: a commentary. PMID- 17124765 TI - Science, biopolitics and risk: margins of uncertainty. PMID- 17124766 TI - Environmental protection and uncertainty: the case of deliberate release of genetically engineered organisms. PMID- 17124767 TI - Taming the technological imperative: a comment. PMID- 17124768 TI - Commentary on Kawar and Sherlock. PMID- 17124769 TI - The ethics of confidentiality: introduction. AB - This essay aims to briefly summarise the collection of articles on confidentiality issues in psychology, and to highlight apparently conflicting opinions about the confidentiality rule. Conflicts are then analysed in terms of competing systems of ethics. Finally, the role for ethics education in psychology education and training is considered. PMID- 17124770 TI - Confidentiality in psychological practice. AB - Confidentiality has long been a cornerstone of trust in the professional relationship between psychologists and their clients. Developments in computer technology, litigation, insurance reimbursement schemes, and changing lifestyles are forcing psychologists to reconsider and refine their approach to respecting this important ethical principle. This article review basic concepts on the matter, and discusses these in light of evolving issues in practice, technology, and the law. Some contrasts in legal and ethical aspects of confidentiality between Australia and the United States are discussed. Recommendations for enhancing attention to confidentiality in one's practice are included. PMID- 17124772 TI - Fostering proteomics in India. PMID- 17124771 TI - Confidentiality in psychological practice: a decrepit concept? AB - Although the principle of confidentiality in the relationship between psychologists and client has been vaunted, and is emphasised in the Australian Psychological Society's Code of Professional Conduct (the APS code; 1994), the confidentiality of this relationship is circumscribed by the absence of legal protections, the ethical beliefs of psychologists, institutional practices, and the provisions of the APS code itself. Lack of privilege in judicial proceedings, and statutory obligations to report certain types of behaviour, mandate breaches of confidentiality in some circumstances. Ethical beliefs of psychologists may support disclosure, especially where it is believed that there is danger of serious physical harm to the client or others. Multidisciplinary teams and institutional settings require the exchange of information for optimal delivery of services. Recent amendments to the APS code may require disclosure without the client's consent when a client is believed to be suicidal. Such developments, when considered at all, are typically regarded as exceptions to a general obligation of confidentiality. However, discussion of exceptions presupposes agreement on fundamental principle: the significance of, and rationale for, confidentiality in the psychologist-client relationship. It is argued in this paper that the obligation of confidentiality has been assumed rather than vigorously analysed and empirically explored. A critical examination of this obligation is the most appropriate starting point for the rehabilitation of contemporary principles of confidentiality in the psychologist-client relationship. PMID- 17124773 TI - Profiling the plasma proteome with IMS/MS. PMID- 17124774 TI - Digging for disease markers of sepsis. PMID- 17124775 TI - Practical use of allergenic molecules. PMID- 17124776 TI - Plant-derived food allergens: an overview. PMID- 17124777 TI - Parietaria pollen allergens. PMID- 17124778 TI - Engineering allergens. PMID- 17124779 TI - Ige allergen co-recognition: a new concept or just a new name for cross reactivity? PMID- 17124780 TI - Protein microarrays for the diagnosis of allergic diseases: state-of-the-art and future development. AB - In the emerging field of Proteomics, protein microarrays represent an elegant solution for the simultaneous analysis of the (i) abundance, (ii) function and (iii) interaction of proteins on a system-wide scale. The great power of microarray-based miniature solid-phase immunoassays lies in their potential to investigate in parallel large numbers of analyte pairs while employing only minute amounts of biological sample material (e.g., serum). This has inspired researchers to adopt this approach to the development of novel diagnostic tests with to date published examples in the fields of autoimmune diseases, allergy and cancer. Here, we discuss recent advancements in the development of protein microarrays for the profiling of IgE antibodies in the diagnosis of Type-1 related allergic diseases. PMID- 17124781 TI - Bioinformatics: managing the knowledge on allergens. PMID- 17124782 TI - Immunotherapy with peptides. PMID- 17124783 TI - Update on sublingual immunotherapy treatment regimes. PMID- 17124784 TI - Clinical experience with recombinant pollen allergens. PMID- 17124785 TI - Sublingual vaccines based on wild-type recombinant allergens. AB - Sublingual immunotherapy (SLIT) represents a non invasive alternative to subcutaneous immunotherapy in order to treat type I allergies. Vaccines based on recombinant allergens expressed in a native (i.e. wild-type) configuration, formulated with ad hoc adjuvants designed to target Langerhans cells in the sublingual mucosa should allow to induce allergen-specific regulatory T cells. In this context, we have developed animal and human preclinical models to test the capacity of candidate vaccines to modulate selectively allergen-specific T helper lymphocyte polarization following sublingual vaccination. PMID- 17124786 TI - The costs of war and cookie dough. PMID- 17124787 TI - Obstructive sleep apnea and cognitive dysfunction. PMID- 17124788 TI - Diagnosis and management of chronic constipation. PMID- 17124789 TI - The patient with cold hands: understanding Raynaud's disease. AB - The availability of new treatments for Raynaud's disease, as with any medical condition, depends on how convincing the results of clinical research prove to be. The validity and reliability of research in Raynaud's disease is subject to two major constraints: the seasonal and intermittent nature of the condition limits the amount of time patients can be studied continuously, and researchers have failed to agree on objective outcome measures. Vascular physiology is, and no doubt is destined to remain, a dynamic and technology-driven clinical realm. Despite the wide array of promising treatments, the best and most basic management of Raynaud's disease seems to be behavioral and at least partly pharmaceutical. The two biggest behavioral factors are nicotine use and exposure to cold environments. Giving up nicotine can be a daunting challenge for long term smokers and chewing-tobacco users. Avoiding cold environments may be easier said than done, especially for patients who work outdoors or in air-conditioned spaces. Perhaps the best treatment for Raynaud's disease is a reliable diagnosis and the positive prognosis that comes with it. Most patients have a stable course, and nearly half will actually improve with time and steady reassurance. PMID- 17124790 TI - Botulism: rare, but deadly. PMID- 17124791 TI - An endovascular approach to the treatment of intracranial aneurysms. PMID- 17124792 TI - Plain films show the most common hip problem of adolescence. PMID- 17124793 TI - Dramatic signs with cranial nerve deficit. PMID- 17124794 TI - Case of the month. Chronic hepatitis. PMID- 17124795 TI - [Partial lease squares approach to functional analysis]. AB - We extend the partial least squares (PLS) approach to functional data represented in our models by sample paths of stochastic process with continuous time. Due to the infinite dimension, when functional data are used as a predictor for linear regression and classification models, the estimation problem is an ill-posed one. In this context, PLS offers a simple and efficient alternative to the methods based on the principal components of the stochastic process. We compare the results given by the PLS approach and other linear models using several datasets from economy, industry and medical fields. PMID- 17124796 TI - [Various methods of dynamic functional analysis in human sciences and economics]. AB - Including the temporal and developmental dimension into the measurement of human conduct is a fundamental concern for those who do research in natural surroundings. Observing an individual day after day may possibly give a more complete vision of how behavior works than measuring a group of individuals at a single time and analyzing the differences found among them. Unfortunately most of the tools allowing analyzing individual time series call for large numbers of repeated observations. Thus, practicable longitudinal research designs often do not involve either enough repeated measurements for traditional time series analyses nor either replicate enough individuals for traditional, large-sample analyses. Dynamic factor analysis is a rationale and procedure for both pooling relatively short time series information across limited numbers of participants and analyzing the pooled information for its dynamic, process-relevant elements. It is a merging of two important analytical tools - multivariate time series and the common factor model, from which it distinguishes itself mainly by the fact that in dynamic factor analysis, the values of the common factors can influence the values of the observed variables both concurrently and in delayed fashion. Dynamic factor analysis is actually a method which allows detecting structures in the time series as well as the relations between the series and the explanatory variables. We illustrate the different models used in psychology and social sciences, as well as in econometry and economics. PMID- 17124797 TI - The future of dynamic factor analysis in psychology and biomedicine. AB - The currently dominant approach to statistical analysis in psychology and biomedicine is based on analysis of inter-individual variation. Differences between subjects drawn from a population of subjects provide the information to make inferences about states of affairs at the population level (e.g., mean and/or covariance structure). Recently it has been shown that in general the inferred states of affairs at the population level do not apply at the level of intra-individual variation characterizing the life trajectories of individual subjects making up the population. This is a direct consequence of the so-called classical ergodic theorems of Birkhoff and Wiener which has important implications for the way in which psychological and biomedical processes have to be analyzed. The classical ergodic theorems are introduced below in order to show the necessity of using an alternative approach which is valid for the analysis of intra-individual variation. This approach has to be based on single-subject time series analysis. Next an overview is presented of dynamic factor models for the analysis of multivariate time series and the various ways to fit these models to the data. We then turn to an empirical application of factor analysis of personality data obtained in a replicated time series design, showing substantial heterogeneity in intra-individual factorial personality structure. The next topic is entirely innovative--for the first time I present my new dynamic factor model for the analysis of nonstationary time series. In the conclusion I will sketch some biomedical research initiatives in which this new model will be used. PMID- 17124798 TI - [Identification of types in small samples with the help of bootstrap simulation (illustrated with the help of the anxiety/depression subscales of the HAD)]. AB - Using a HAD-file as an illustration, we could show that small files (n = 50) can provide statistically significant results if we apply bootstrap simulation. First the subscales "anxiety" and "depression" are computed and afterwards classified (in three categories). Both classified subscales are cross tabled and submitted to a configurational cluster analysis. One type can be clearly identified. To test the stability of the one-type solution, a bootstrap simulation (Lautsch/von Weber: BOOTSTRAP) is applied. The simulation confirms the one-type solution. PMID- 17124799 TI - [Socio-demographic identification of anxiety-depression types (socio-demographic specification and prediction of types)]. AB - In the article "Identification of Types in Small Samples with the help of Bootstrap Simulation" we could show how anxiety-depression types could be explored in small samples. We used the two sub-scales (anxiety and depression) in accordance with the indications of the HAD manual of Zigmond, A.S. & Snaith R.P. . The extraction of types will be repeated with the data of n=168 homeless people. Then the socio-demographic homogeneity of the extracted types will be tested. With this procedure we want to answer the question, whether in the sub group of homeless people, the knowledge the degree of expression of some significant socio-demographic traits will allow us assigning them to specific anxiety-depression types. PMID- 17124800 TI - [Explanation and forecast: relapse of juvenile offenders]. AB - On the basis of n=82 juvenile offenders from a prison for juvenile offenders in Rheinland Pfalz the model of the logistic regression is compared with a procedure from the family of the neural nets in its efficiency to explain and predict "relapse" in form of a renewed imprisonment or prosecution /police search after dismissal. The group which can be examined is limited by the population of the prison for juvenile offenders and the explaining variables for "relapse" as "addicted to drugs" present non-metric scaling. For the explanation only probabilities for "relapse" can be indicated in this connection. By means of this probability it is possible to classify the individual case. The forecast is simulated by coincidental dividing of the data: the first part of the data is used for the explanation, the second for the forecast. With the comparison of the logistic regression with the neural nets, the superiority of neural nets in the explanation of "relapse" can be shown, since the neural nets are able to consider dependence between the explaining variables and according to that they offer a differentiated explanation. Their efficiency to predict "relapse" depends on the comparability of the distribution in the two coincidentally provided samples, the training data record for determining the explanation and the test case for the use of the explanation regarding the forecast. For optimal explanation and forecast neural nets are to be preferred to the logistic regression, since in the model with the better explanation also includes the potential for a usable better forecast. Moreover the model of the logistic regression is in fact a special case of the neural net, with a reduced complexity of the net. PMID- 17124801 TI - [Importance and extension of the qualitative approach in phenomeno-structural method in psychopathology]. AB - Fundamental principles of phenomeno-structural method partly originate from a phenomenological approach in psychiatry for which, as Jaspers says, "the important thing is less the study of innumerable cases than the intuitive and major comprehension of some particular ones". It means that, far from traditional statistical proceedings, it does not give any priority to the temptation of a cumulative and accumulative collection of data, for which it even receives their greatest misgivings, leaving it out of its procedures. This method also owes a lot to Bergson's insistence on the qualitative characteristics of the "immediate data of consciousness" and of experienced temporality, for which analytical and chronological fragmentations can do nothing but distort the authentically comprehensive grasping, and make them loose their specificity, unity and thus the indecomposability that, by nature, they precisely contain. According to Eugene Minkowski, the psychopathical break-up, proceeding from the deep heart of life, is essentially going to express itself, with its own properties, as a qualitative modification. In this way, the study of some cases carefully chosen, in accordance with their "typical" value, is more important than a blind acquisition of data, i.e. without previous option, orientation or perspective. Starting from their spontaneous words collected during a conversation, the meticulous analysis of the language of the patients, put in resonance with the vivid metaphors of the language in which they are expressed, as Minkowski will so brilliantly show, or via mediations connecting image and language, as in the experiment of the Rorschach--particularly approached in its essential mechanisms by his wife Francoise Minkowska--represents, from this point of view, a mediator and an irreplaceable instrument of a qualitative approach of a normal as well as a pathological personality and, more widely, of the person itself. PMID- 17124802 TI - [Rating scales based on the phenomenological and structural approach]. AB - A current tendency of research in clinical psychology consists in using an integrated quantitative and qualitative methodology. This approach is especially suited to the study of the therapeutic intervention where the researcher is himself part of the situation he is investigating. As to the tools of research, the combination of the semi-structured clinical interview, of psychometric scales and projective tests has proved to be pertinent to describe the multidimensional and fluctuating reality of the therapeutic relationship and the changes induced by it in the two partners. In arts therapeutic research the investigation of the artistic production or of the free expression of people may complete the psychometric and projective tools. The concept of "expressive test" is currently being used to characterise this method. In this context, the development of rating scales, based on the phenomenological and structural or holistic approach allows us making the link between qualitative analysis and quantification, leading to the use of inferential statistics, providing that we remain at the nominal or ordinal level of measurement. We are explaining the principle of construction of these rating scales and we are illustrating our practice with some examples drawn from studies we realized in clinical psychology. PMID- 17124803 TI - [Using fairy tales and narrative strategies with the help of a sandpit with children suffering from conduct disorders]. AB - Our research, focussed on art therapy with primary school children, guided us to the footsteps of women pioneers in the research about children's psyche, like Anna Freud, Melanie Klein, Margaret Loewenfeld, Dora Kalff. We have been inspired by their very personal but nevertheless somehow similar type of research, to work with the children in a blue sand pit, where they could construct and play their personal fairy tales. Thus they share with us their momentary preoccupations, without being forced to talk about them. They are revealing their internal structures, and are integrating, by narration and projection on those figurines in action, their deficient defense mechanisms as well as their negative tendencies. Based on this approach, we expect that there will by an amelioration of their social skills, an enhancement of their intrinsic motivation, and a diminution of their conduct disorders. The analyze will be done by questionnaires and rating scales constructed especially for this research, as well as by sociograms, with the help of descriptive non parametric statistics. PMID- 17124804 TI - [Analysis of pictures of depressive patients from the point of view of mood and emotions]. AB - The question to what extent pictures drawn by persons suffering from depression can provide clues as to the syndrome or the stage of the illness, has been since long an intensively discussed subject of research. The following article is providing some insight into the investigation of pictures drawn by depressive patients and stresses some research results. According to the statistical evaluation, the analysis of pictures, by means of an observational frame, shows significant correlations with the Hamilton Test. PMID- 17124805 TI - [Multidimensional homogeneity analysis with small samples: comparison of different subgroups of people suffering from exclusion]. AB - With the aim of studying the relationship between harmful biographical events and exclusion we have developed a rating scale for semi-structured biographical interviews. The rating scale was constructed in accordance with the phenomenological and structural approach and allows us gathering some clinically pertinent indicators concerning the course of life, the nature of the trauma, the composition of the original family, the life period of emotional pain and the current familiar and social support. The homogeneity or multiple correspondence analysis (HOMALS) led to the extraction of the fundamental dimensions underlying the data of three clinical subgroups of people suffering from exclusion and marginalization: homeless and long term unemployed people, immigrants, political refugees and asylum seekers, drug addicts. The results demonstrate the relevance of the multidimensional homogeneity analysis applied to this kind of data. On one hand they suggest a theoretical model of the relationship between harmful biographical events, vulnerability, exclusion and psychopathological complications. On the other hand they show that we are facing differential psychological profiles which will be studied thoroughly during the next stages of our research. PMID- 17124806 TI - "Statistical evidence" for the investigation of international crimes. AB - Statistics are widely applied throughout scientific research disciplines but 'statistical evidence' sounds surely an unhappy term rather avoided. In International Criminal Law the term seems to stress the value that statistical methods have for the analysis of large-scale victimisation and describes rather the tools used to support the evidence. However, statistics is required for valid, reliable and objective results that would otherwise be left in the dusty political sphere of the warring parties and their actors. But what are the concepts to do so? Today's presentation shows two widely known cases from the International Criminal Tribunal for the Former Yugoslavia (ICTY) where statisticians analysed the Crime Scenes in Srebrenica and Kosovo. The cases introduce two different analytical approaches. Firstly, in Srebrenica the analysts focussed on civilians that were killed during the take over of the enclave. Thus the findings are descriptive, summarizing observations and presenting results based on simple statistical tools. Secondly, in Kosovo the researchers asked what caused the killings and refugee migration during the NATO air campaign in springtime 1999. Here, in a complex environment statistical methods are exercised to determine causal relations. Hypotheses are tested to be consistent with huge data sets collected from a broad field of sources, to finally end up with Statistical Evidence. PMID- 17124807 TI - Home, sweet dental home. PMID- 17124808 TI - The ethics of esthetic dentistty: a neglected topic. PMID- 17124809 TI - Porcelain laminate veneers in the adult dentition. PMID- 17124810 TI - Porcelain veneers--preliminary procedures before preparation. PMID- 17124811 TI - Porcelain veneers: try-in, cementation and maintenance. PMID- 17124812 TI - Maximizing the esthetics of anterior teeth by conservative restorative dentistry: resin-based composite veneers. PMID- 17124813 TI - The significance of cuspid guided occlusion. PMID- 17124814 TI - Things to know about dental scrap metal. PMID- 17124815 TI - Oral and maxillofacial pathology case of the month. Erosion secondary to bulimia nervosa. PMID- 17124816 TI - Determining venous incompetence: a report from a specialised leg ulcer clinic. AB - OBJECTIVE: To analyse the diagnosis, treatment and prognosis in patients attending a specialised leg ulcer clinic at a dermatology department. METHOD: In total, 345 patients were investigated and 332 registered and followed up prospectively. All patients had their arterial and venous circulation assessed with a hand-held Doppler ultrasound. RESULTS: The most frequent diagnosis was venous ulceration (153 patients, 46%) followed by hydrostatic ulceration (70 patients, 21%). Venous incompetence was classified as isolated superficial (n=86) or deep venous incompetence (n=57) in 143 out of the 153 patients. Previous deep vein thrombosis (DVT) was more frequent in patients with deep venous incompetence. Of patients with venous ulcers, 38 (25%) healed within 92 days, 77 (50%) within 155 days and 115 (75%) within 329 days. Healing time was influenced by patient age, ulcer duration and ulcer area, but not by type of venous incompetence or ankle brachial pressure index. After healing, 19% of venous patients (28/144), dominated by those with superficial disease, were subject to venous vascular surgery. CONCLUSION: Classification of venous insufficiency should be mandatory in patients with venous ulcers since it determines suitability for venous surgery. PMID- 17124817 TI - Reviewing the evidence for wound bed preparation. AB - This paper explores the nature of evidence and how it has evolved in recent years, and sets out a process for assembling and assessing the evidence to support wound bed preparation as an effective method of managing chronic wounds PMID- 17124818 TI - Conservative debridement of postoperative Meleney's gangrene following a caesarean section. AB - Meleney's gangrene is almost exclusively postoperative and is caused by multiple bacteria. This case review describes the conservative management of Meleney's gangrene in a young patient following a Caesarean section. PMID- 17124819 TI - An evaluation of polyvinylidene film dressing for treatment of pressure ulcers in older people. AB - OBJECTIVE: To compare the effectiveness of polyvinylidene (PVL) food wrap as a dressing material versus conventional ointments and gauze dressings for pressure ulcers in patients aged 60 years or over. METHODS: A prospective open controlled trial was conducted. The severity of pressure ulcers in both groups was evaluated using the DESIGN system. Scores were compared at baseline, four, eight and 12 weeks. RESULTS: Twenty-five patients were treated with the PVL film (test) dressings and 24 with the conventional treatment (control). The test group showed greater improvement in DESIGN scores than the control group throughout the observation period, and the difference reached statistical significance at 12 weeks (p < 0.05). The incidences of local wound infection in the two groups were not statistically different (p > 0.999). CONCLUSION: The PVL film dressing was more effective than conventional treatment in the management of severe pressure ulcers. PMID- 17124820 TI - A prospective study: hyperbaric oxygen therapy in diabetics with chronic foot ulcers. AB - Research suggests that hyperbaric oxygen therapy may have beneficial effects on ulcer healing and amputation rates in diabetic patients. This paper describes the design of a study that is evaluating its effects on chronic diabetic foot ulcers. PMID- 17124821 TI - Validity of subgroup comparisons in real-life studies. PMID- 17124822 TI - Cost of managing chronic wounds in the U.K., with particular emphasis on maggot debridement therapy. AB - The importance of debridement in wound healing is well established. This review calculates the cost savings that might be achieved when using maggot debridement therapy on chronic wounds instead of autolytic debridement. PMID- 17124823 TI - Efficacy of a skin-protection powder for use as a dressing for intractable ulcers. AB - OBJECTIVE: Stomahesive skin-protection powder has been reported to be useful as a skin-care and skin-barrier product for the management of stomas. This study aimed to evaluate its efficacy, in terms of wound healing, moisture retention and pain management, as an alternative to conventional dressing materials. Both clinical and animal studies were undertaken. METHOD: The efficacy of the Stomahesive powder was tested by measuring the thickness of granulation tissue formed in a total skin defect in a db/db mouse model. We then compared the healing process using either the skin-protection powder or a conventional film dressing material. In the clinical study 17 patients with various intractable ulcers were treated with Stomahesive powder, and healing was evaluated. RESULTS: In the mouse model, granulation tissue in the wounds treated with the powder was 2.86 times thicker than that of the wounds treated with the film dressing. In the clinical study, 16 out of 17 wounds healed completely. CONCLUSION: The Stomahesive powder could be an effective treatment modality for contact ulceration, superficial ulcers with complex contours and morphology, and superficial ulcers contaminated by liquid faeces or vaginal discharge that have not responded to conventional dressings. DECLARATION OF INTEREST: None. PMID- 17124824 TI - Research in wound healing and tissue repair--whose baby is it? AB - New developments in wound care are most likely to be implemented if they result from multidisciplinary collaboration. Central to such success is communication, through journals and at conferences, of research projects while still in progress PMID- 17124826 TI - "One mighty database from cradle to grave gives me the creeps". PMID- 17124825 TI - Low demand for homeowner help. PMID- 17124827 TI - "They saw that only an empowered workforce can empower its patients". PMID- 17124828 TI - Why is mental health care still in crisis? PMID- 17124829 TI - An independent mind. PMID- 17124830 TI - The implications of reusing single-use medical devices. AB - The Medicines and Healthcare products Regulatory Agency (MHRA) has updated its advice on the implications of reusing single-use medical devices. This article discusses what constitutes a single-use device, the implications of reuse and the legal and safety issues. PMID- 17124831 TI - Chest examination part 2--chest percussion. PMID- 17124832 TI - Fluid-balance monitoring, part of track and trigger. AB - This article (part four of four) explores urine output and fluid management, related conditions and nursing care. PMID- 17124833 TI - Confidentiality and protection when working with under-16s. AB - When a condom registration scheme was set up at a sexual health clinic to address child protection concerns it resulted in a reduction in the number of young people using the service. The scheme was abandoned and a pilot was carried out to assess young people's views on condom provision. This article reviews the pilot and discusses issues of confidentiality versus child protection when working with under-16s. This is a summary of the paper: the full version can be accessed at nursingtimes.net. PMID- 17124834 TI - Communicating the risks of hepatitis B to travellers. PMID- 17124835 TI - Advanced practitioners' use of reflexivity in decision-making. AB - This small study provides an insight into the process of clinical decision-making and raises awareness of the role of reflexivity and intuition in expert practice. This is a summary of the paper--the full version can be accessed at: nursingtimes.net. PMID- 17124836 TI - "The nonsense that is the private finance initiative". PMID- 17124837 TI - Effects of unilateral laser-assisted ventriculocordectomy in horses with laryngeal hemiplegia. AB - REASONS FOR PERFORMING STUDY: Recent studies have evaluated surgical techniques aimed at reducing noise and improving airway function in horses with recurrent laryngeal neuropathy (RLN). These techniques require general anaesthesia and are invasive. A minimally invasive transnasal surgical technique for treatment of RLN that may be employed in the standing, sedated horse would be advantageous. OBJECTIVE: To determine whether unilateral laser-assisted ventriculocordectomy (LVC) improves upper airway function and reduces noise during inhalation in exercising horses with laryngeal hemiplegia (LH). METHODS: Six Standardbred horses were used; respiratory sound and inspiratory transupper airway pressure (Pui) measured before and after induction of LH, and 60, 90 and 120 days after LVC. Inspiratory sound level (SL) and the sound intensities of formants 1, 2 and 3 (Fl, F2 and F3, respectively), were measured using computer-based sound analysis programmes. In addition, upper airway endoscopy was performed at each time interval, at rest and during treadmill exercise. RESULTS: In LH-affected horses, Pui, SL and the sound intensity of F2 and F3 were increased significantly from baseline values. At 60 days after LVC, Pui and SL had returned to baseline, and F2 and F3 values had improved partially compared to LH values. At 90 and 120 days, however, SL increased again to LH levels. CONCLUSIONS: LVC decreases LH associated airway obstruction by 60 days after surgery, and reduces inspiratory noise but not as effectively as bilateral ventriculocordectomy. POTENTIAL RELEVANCE: LVC may be recommended as a treatment of LH, where reduction of upper airway obstruction and respiratory noise is desired and the owner wishes to avoid risks associated with a laryngotomy incision or general anaesthesia. PMID- 17124838 TI - Post anaesthetic myopathy/neuropathy in horses undergoing magnetic resonance imaging compared to horses undergoing surgery. AB - REASONS FOR PERFORMING STUDY: Patient positioning and long anaesthetic duration required for magnetic resonance imaging (MRI) may result in a higher frequency of post anaesthetic myopathy/neuropathy syndrome (PAMNS) as compared to horses undergoing anaesthesia for surgery. HYPOTHESIS: Equine anaesthesia for MRI is associated with a higher frequency of PAMNS than anaesthesia for nonemergency, nonabdominal surgery. METHODS: Anaesthetic and medical records of horses (n = 633) undergoing MRI or surgery between January 2001 and January 2005 (inclusive), were reviewed. Information obtained included patient details (breed, sex, age, bodyweight), area of body scanned or involved in surgery, body position, anaesthetic and inotropic agents administered, anaesthetic duration, adverse events during anaesthesia and outcome at 7 days. Data were examined by cross tabulation and Chi-square or Fisher's exact test of association. The influence of individual variables was examined by univariant and multivariant analysis models. RESULTS: There were no statistically significant differences between the 2 groups in parameters examined, except that horses in the MRI group were heavier (P<0.0001) and anaesthetic duration longer in the surgery group (P<0.004). Eight horses (2.3%, 95% Confidence interval [CI]: 1.1-4.2) in the MRI group had clinical signs of PAMNS in the post anaesthetic period, whereas only 2 horses (0.98%, 95% CI: 0.2-2.8) in the surgery group were affected. This was not statistically significantly different (odds ratio = 2.7, 95% CI: 0.8-13, P = 0.3). Two horses undergoing MRI were subjected to euthanasia due to the severity of PAMNS. CONCLUSIONS: There was no difference in the occurrence of PAMNS between the 2 groups. POTENTIAL RELEVANCE: The risk of performing general anaesthesia for diagnostic procedures such as MRI may not be greater than that for a surgical procedure. However, the benefits should be carefully weighed against the risks involved. PMID- 17124839 TI - Cartilage oligomeric matrix protein and hyaluronan levels in synovial fluid from horses with osteoarthritis of the tarsometatarsal joint compared to a control population. AB - REASONS FOR PERFORMING STUDY: Quantification of cartilage oligomeric matrix protein (COMP) levels within synovial fluid from the tarsometatarsal joint has not previously been reported and an effective synovial fluid marker would allow monitoring of disease progression and treatment. OBJECTIVES: To quantify levels of COMP and hyaluronan (HA) in synovial fluid from the tarsometatarsal joint, identify differences in levels from horses with osteoarthritis (OA) of the tarsometatarsal joint compared to a control population and to correlate levels with radiographic changes in horses with OA. METHODS: Synovial fluid was collected from the tarsometatarsal joint of 25 horses without hindlimb lameness (controls) and 25 lame horses, subjected to analgesia of the joint. COMP concentrations were measured using a homologous inhibition ELISA. Immunoblots of synovial fluid from 3 lame horses and 3 controls were performed to identify fragmentation of COMP. Hyaluronan (HA) concentration in synovial fluid was determined using a competition ELISA. Radiographs of the lame horses with OA were scored and correlated with levels of COMP and HA. RESULTS: Concentrations of COMP in OA of the tarsometatarsal joint were significantly lower than in the control samples. An additional fragment band of COMP (approximately 30 kDa) was identified on the immunoblots of the horses with OA and this fragment was not identified in controls. No significant difference was identified in the HA or HA:COMP ratio between lame and control horses. There was no correlation between levels of synovial fluid COMP and HA, and radiographic changes. CONCLUSIONS AND POTENTIAL RELEVANCE: Lowered levels of COMP in synovial fluid of tarsometatarsal joints correlates with the presence of osteoarthritis. However, a single value cannot be used to stage the disease process. Levels of HA may not be a useful marker for this disease. Decreased, rather than increased COMP levels, may reflect significant loss of cartilage in established osteoarthritis. A specific assay for the COMP fragment generated with osteoarthritis may allow the earlier detection of clinical cases. PMID- 17124840 TI - Magnetic resonance imaging, ultrasonography and histology of the suspensory ligament origin: a comparative study of normal anatomy of warmblood horses. AB - REASONS FOR PERFORMING STUDY: The diagnosis of lameness caused by proximal metacarpal and metatarsal pain can be challenging. Magnetic resonance imaging (MRI) offers the possibility for further diagnosis but there have been no studies on the normal MRI appearance of the origin of the suspensory ligament (OSL) in conjunction with ultrasonography and histology. OBJECTIVES: To describe the MRI appearance of the OSL in fore- and hindlimbs of sound horses and compare it to the ultrasonographic and histological appearance. The findings can be used as reference values to recognise pathology in the OSL. METHODS: The OSL in the fore- and hindlimbs of 6 sound horses was examined by ultrasonography prior to death, and MRI and histology post mortem. Qualitative evaluation and morphometry of the OSL were performed and results of all modalities compared. RESULTS: Muscular tissue, artefacts, variable SL size and shape complicated ultrasonographic interpretation. In MRI and histology the forelimb OSL consisted of 2 portions, the lateral being significantly thicker than medial. The hindlimb SL had a single large area of origin. In fore- and hindlimbs, the amount of muscular tissue was significantly larger laterally than medially. Overall SL measurements using MRI were significantly higher than using histology and ultrasonography and histological higher than ultrasonographic measurements. Morphologically, there was a good correlation between MRI and histology. CONCLUSIONS: MRI provides more detailed information than ultrasonography regarding muscle fibre detection and OSL dimension and correlates morphologically well with histology. Therefore, ultrasonographic results should be regarded with caution. POTENTIAL RELEVANCE: MRI may be a diagnostic aid when other modalities fail to identify clearly the cause of proximal metacarpal and metatarsal pain; and may improve selection of adequate therapy and prognosis for injuries in this region. PMID- 17124841 TI - Continuous monitoring of penicillin G and gentamicin in allantoic fluid of pregnant pony mares by in vivo microdialysis. AB - REASONS FOR PERFORMING STUDY: Most current treatments for placentitis in mares are empirical with few control studies to evaluate their effectiveness. OBJECTIVE: To monitor drug concentrations in allantoic fluid of pregnant pony mares using in vivo microdialysis and establish if this method would be useful for determining allantoic concentrations of drugs in normal mares and those with placentitis. METHODS: Five late gestational pony mares had microdialysis probes inserted into the allantoic fluid using transabdominal ultrasound-guided allantocentesis. Single injections of penicillin G (22,000 u/kg), gentamicin (6.6 mg/kg bwt) and flunixin meglumine (1 mg/kg bwt) were administered i.v. and dialysate samples collected continuously for 24 h. In a separate study, drug concentrations were monitored in allantoic fluid of 2 mares with experimental placentitis induced by intracervical inoculation with Streptococcus equi ssp. zooepidemicus. Drug concentrations were measured by high performance liquid chromatography (penicillin G, flunixin meglumine) or enzyme-linked immunosorbent assay (gentamicin). RESULTS: Penicillin G and gentamicin achieved average peak concentrations of 9.8+/-2.2 and 8.5+/-3.1 microg/ml, respectively, in allantoic fluid of noninfected mares. Pharmacokinetic comparisons indicate that penicillin G persists much longer in allantoic fluid than blood, whereas gentamicin exhibited similar profiles in the 2 compartments. Flunixin meglumine was not detected in allantoic fluid. In infected mares, penicillin G achieved a similar peak concentration in allantoic fluid (11.2 microg/ml) whereas peak gentamicin concentration (3.9 microg/ml) appeared to be reduced relative to drug concentrations in noninfected mares. CONCLUSIONS: Microdialysis is a useful technique for continuous in vivo monitoring of drugs in equine allantoic fluid. Our results indicate that penicillin G and gentamicin undergo effective placental transfer in pregnant mares and in 2 mares that transplacental drug transfer may be altered selectively if active placental infection is present. POTENTIAL RELEVANCE: Further studies are needed to evaluate the feasibility of using increased dose intervals for penicillin G and an increased dose rate of gentamicin to effectively combat placental infections in mares. PMID- 17124842 TI - Are feeding practices associated with duodenitis-proximal jejunitis? AB - REASONS FOR PERFORMING STUDY: Feeding concentrate has been putatively associated with risk of development of duodenitis-proximal jejunitis (DPJ); however, this association has not been evaluated systematically in a controlled study. OBJECTIVES: To determine whether there was evidence that feeding practices were associated with increased odds of developing DPJ employing a case control study. HYPOTHESIS: The amount of concentrate fed daily to horses is significantly greater among horses that develop DPJ than control horses with either lameness or other types of colic. METHODS: Feeding practices of cases of DPJ diagnosed between 1997 and 2003 were compared with those of 2 populations of control horses (colic controls and lameness controls) admitted to the clinic from the same time period. Following multiple imputation of missing data, comparisons were made using polytomous logistic regression. RESULTS: Horses with DPJ were fed significantly more concentrate and were significantly more likely to have grazed pasture than either control populations; DPJ horses were significantly more likely to be female than were lameness horses. Results were unchanged after adjusting for bodyweight of the horse. CONCLUSIONS: Feeding and grazing practices differ among horses with DPJ relative to horses with other forms of colic and lame horses. POTENTIAL RELEVANCE: The observed magnitudes of association were not sufficiently strong to merit diagnostic/predictive application; however, these observations, if substantiated by other studies, might provide important aetiological clues. PMID- 17124843 TI - Growth factor and receptor mRNA expression in the intestine of horses with large colon volvulus: a pilot study. AB - REASONS FOR PERFORMING STUDY: Growth factors (GF) are important for maintenance and repair of intestinal mucosal structure and function, but there have been no studies investigating growth factor (GF) or growth factor receptor (GF-R) mRNA expression in the intestine of horses with large colon volvulus (LCV). OBJECTIVES: (1) To determine mRNA expression for epidermal growth factor (EGF), EGF receptor (EGF-R), insulin-like growth factor-I (IGF), IGF receptor (IGF-R), vascular endothelial growth factor (VEGF) and VEGF receptor (VEGF-R) in the intestine of horses with an LCV compared to normal intestine. (2) To measure the correlation between histological intestinal injury and mRNA expression. METHODS: In 5 horses, samples were collected from the mid-jejunum (small intestine, SI), pelvic flexure (PF) and right dorsal colon (RDC) prior to creation of the LCV (NORM), 1 h following creation of the LCV (ISCH) and 1 h following correction of the LCV (REPER). In 2 clinical cases of LCV, samples were collected from the PF and RDC. Samples were assessed histologically for the amount of intestinal injury. The mRNA expressions of growth factors and receptors were determined using qRT-PCR. RESULTS: VEGF and VEGF-R mRNA expression was greater in horses with an LCV compared to NORM. Expression of IGF-R mRNA increased in the SI during ISCH and REPER. CONCLUSION AND POTENTIAL RELEVANCE: The increase compared to NORM in VEGF and VEGF-R mRNA expression in horses with LCV may be important in early intestinal healing and may also explain, in part, the increase in vascular permeability in horses with a LCV. Expression of IGF and IGF-R in the SI warrants further investigation and may be important for understanding post operative complications in horses with SI lesions. PMID- 17124844 TI - Proximal interphalangeal joint arthrodesis using a combination plate-screw technique in 53 horses (1994-2003). AB - REASONS FOR PERFORMING STUDY: A method for proximal interphalangeal joint (PIP joint) arthrodesis that provides a stable fixation and minimal duration of cast support is evaluated retrospectively. OBJECTIVES: Evaluate the clinical use of a combined plate-screw method for PIP joint arthrodesis in a large number of horses. METHODS: The records of 53 horses undergoing PIP joint arthrodesis were reviewed. Arthrodesis was performed with a dorsally placed 3-, 4- or 5-hole narrow dynamic compression plate (DCP) with 2 transarticular cortex screws placed in lag fashion either side of the plate. Subject details, clinical presentation, radiographic findings, surgical technique, post operative treatment and complications were recorded. Long-term follow up (mean 3 years) was obtained for 46 horses. RESULTS: Arthrodesis procedures (n = 58) were performed on 53 horses with a DCP in combination with transarticular cortex screws placed in lag fashion. Conditions treated were osteoarthritis (OA) of the PIP joint, fracture of middle phalanx, PIP joint subluxation, subchondral cystic lesions and degenerative joint disease secondary to sepsis. Time of post operative cast application was 14 days. Overall 40/46 (87%) horses could be used as intended including 20/25 (81%) forelimb and 20/21 (95%) hindlimb arthrodeses. Twenty-three of 27 (85%) horses used for performance had successful outcomes. Complications included implant infection, cast sores and partial implant failure. CONCLUSIONS: PIP joint arthrodesis using a DCP and transarticular cortex screws placed in lag fashion provides a stable construct and short casting period with minimal complications. The prognosis for return to performance was excellent for horses treated with hindlimb PIP joint arthrodesis and good for forelimb arthrodesis. POTENTIAL RELEVANCE: Use of a combination technique for PIP joint arthrodesis allows a high proportion of horses with pastern joint disease to be returned to their athletic potential. PMID- 17124845 TI - Factors influencing pre-race serum concentration of total carbon dioxide in Thoroughbred horses racing in California. AB - REASONS FOR PERFORMING STUDY: Many racing jurisdictions monitor pre-race serum concentration of total carbon dioxide (TCO2) among racing horses. To our knowledge, factors influencing concentration of TCO2 among horses participating in racing have not been systematically evaluated and reported. OBJECTIVES: To determine if characteristics of horses and racing conditions routinely recorded were significantly associated with pre-race concentration of TCO2, while accounting for and estimating effects of trainer and horse. METHODS: Pre-race serum TCO2 concentrations from 5028 starts made by 2,349 horses trained by 287 trainers at 2 racetracks in California during 2005 were examined. Data regarding characteristics of starters and race conditions obtained from a commercial database were recorded for each start. Data were analysed using mixed-effects, with TCO2 concentration as the dependent variable, and trainer and horse nested within trainer as random effects. RESULTS: Sex, class and distance of race, frusemide administration and cloudy weather conditions were significantly (P<0.001) associated with pre-race TCO2 concentration. Horses that finished in the top 3 positions had values that were slightly (0.2 mmol/) but significantly (P<0.001) greater than horses not finishing in the top 3. There were significant effects of trainer on pre-race TCO2 concentration. CONCLUSIONS: A variety of factors may influence pre-race TCO2 concentration in horses. Horses with better performance tend to have higher pre-race TCO2 concentrations. POTENTIAL RELEVANCE: TCO2 concentration is associated with improved performance although the magnitude of effect was quite small. Regulatory programmes based on monitoring should consider the influence of other factors on TCO2 concentration. PMID- 17124846 TI - Effects of a Streptococcus equi infection--mediated nutritional insult during mid gestation in primiparous Thoroughbred fillies. Part 1: placental and fetal development. AB - REASONS FOR PERFORMING STUDY: There is a paucity of information on factors that influence placental development in the horse. HYPOTHESIS: Changes in nutrition, particularly around the time of proliferative placental growth, can affect development of the placenta and fetal growth. OBJECTIVE: To investigate the effects of 2 planes of nutrition and an unforeseen infection-mediated nutritional insult on placental and fetal development in the mare. METHODS: Twenty maiden Thoroughbred fillies, age 3 or 4 years, mated to one Thoroughbred stallion, were maintained on either High or Moderate food intake throughout pregnancy. In mid gestation all the mares unexpectedly became infected with Streptococcus equi and suffered varying degrees of weight loss as a consequence. Gross and stereological measurements of the placenta and measurements of foal birthweight, crown-rump length, ponderal index and plasma IGF-1 concentration were made at term. RESULTS: Gains in bodyweight during gestation were significantly higher in the High vs. the Moderate nutrition groups. Placental and fetal growth parameters were not influenced by the plane of nutrition. However, transient weight loss in mid gestation from acutely reduced food intake resulted in morphological changes to the allantochorion and decreased foal birthweight. CONCLUSIONS: Excessive weight gain during gestation in maiden Thoroughbred fillies does not result in nutrient partitioning to the dam at the expense of the fetus. However, sudden weight loss in mid-gestation has detrimental effects on placental development which results in reduced fetal growth. POTENTIAL RELEVANCE: Clinicians should appreciate the likely effects of maternal infection and resulting weight loss on pregnancy outcome. PMID- 17124847 TI - Pressure algometry to quantify muscle pain in racehorses with suspected sacroiliac dysfunction. AB - REASONS FOR PERFORMING STUDY: Despite the prevalence of orthopaedic injuries to horses, there is no objective means of quantifying the intensity of musculoskeletal pain. HYPOTHESES: Mechanical nociceptive thresholds (MNT) can be measured repeatably by pressure algometry in horses and MNTs are correlated with both severity of clinical signs and subjective scores of muscle pain on palpation in horses with suspected sacroiliac dysfunction (SID). METHODS: The technique of pressure algometry and its repeatability was tested at 4 anatomical sites on either side of the thoracolumbar and pelvic region in 12 Thoroughbreds in training. In a second series of 15 racing Thoroughbreds, using a different set of landmarks, pain on palpation was assessed by pressure algometry. Horses were grouped based on clinical scores of SID as normal (n = 5), mild (n = 5), moderate (n = 4) and severe (n = 1) suspected SID and scored for muscle pain response by manual palpation. RESULTS: Pressure algometry was shown to be a repeatable measure of MNTs. Horses with suspected SID had significantly lower mean MNT when sites and horses were pooled and showed greater differences in mean algometer measurements between left and right sides, compared to control horses. A significant correlation was found between mean pressure algometry measurements and both suspected SID grade and muscle pain response on palpation. CONCLUSION AND POTENTIAL RELEVANCE: Horses with suspected SID displayed lower MNTs compared to control horses, especially in the pelvic region. This supports a potential role for pressure algometry in providing an objective means of quantifying musculoskeletal pain reflected as a reduced MNT associated with SID and its response to physiotherapy or other treatments. PMID- 17124848 TI - Intravenous and intratracheal administration of trimetoquinol, a fast-acting short-lived bronchodilator in horses with 'heaves'. AB - REASON FOR PERFORMING STUDY: Trimetoquinol (TMQ) is a potent beta-adrenoceptor agonist bronchodilator used in human medicine but has not been evaluated for potential use as a therapeutic agent for horses with 'heaves'. OBJECTIVES: To assess the pharmacodynamics of TMQ in horses with 'heaves' to determine potential therapeutic effects. METHODS: Increasing doses of TMQ were administered to horses with 'heaves' by i.v. and intratracheal (i.t.) routes. Doses ranged 0.001-0.2 microg/kg bwt i.v. and 0.01-2 microg/kg bwt i.t. Cardiac and airways effects were assessed by measurement of heart rate (HR) and maximal change in pleural pressure (deltaPplmax), respectively. Side effects of sweating, agitation and muscle trembling were scored subjectively. Duration of action to i.v. (0.2 microg/kg bwt) and i.t. (2 microg/kg bwt) TMQ was evaluated over 6 h. RESULTS: Intravenous TMQ was an exceptionally potent cardiac stimulant. Heart rate increased at 0.01 microg/kg bwt, and was still increasing after administration of highest dose, 0.2 microg/kg bwt. Airway bronchodilation, measured as a decrease in deltaPplmax, also commenced at 0.01 microg/kg bwt. By the i.t. route, TMQ was 50-100-fold less potent than by i.v. Side effects included sweating, agitation and muscle trembling. Overall, the onset of HR and bronchodilator effects was rapid, within about 3 min, but effects were over at 2 h. CONCLUSION: When administered i.v. and i.t., TMQ is a highly potent cardiac stimulant and a modest bronchodilator. It may not be an appropriate pharmacological agent by i.v. and i.t. routes for the alleviation of signs in horses with 'heaves'. Further studies of TMQ by oral and aerosol routes are necessary. POTENTIAL RELEVANCE: In horses, TMQ is a fast acting bronchodilator with a short duration of action. It could be used as a rescue agent during an episode of 'heaves'. The i.v. and i.t. administration of TMQ is associated with side effects, similar to those reported for all other beta agonists. However, other routes, such as aerosol and oral, may prove useful and safe for the alleviation of bronchoconstriction typical of 'heaves'. PMID- 17124849 TI - Theophylline does not potentiate the effects of a low dose of dexamethasone in horses with recurrent airway obstruction. AB - REASONS FOR PERFORMING STUDY: Theophylline has been shown to have corticosteroid sparing effects for the treatment of human asthma. A similar effect, if present in horses, would allow diminishing the dose of corticosteroids administered to equine patients with inflammatory airway diseases. OBJECTIVES: To evaluate whether theophylline potentiates the effects of a low dose of dexamethasone when treating horses with recurrent airway obstruction (RAO). HYPOTHESIS: Theophylline has steroid-sparing effects in horses with RAO. METHODS: Ten mature mixed breed horses in clinical exacerbation of RAO were studied. Using an incomplete crossover design and 3 experimental periods of 7 days duration, horses were distributed randomly in 5 treatment groups; and administered dexamethasone s.i.d., at either 0.05 mg/kg bwt i.v. or per os, or 0.02 mg/kg bwt alone or combined with theophylline at 5 mg/kg bwt per os b.i.d. A fifth group was treated with theophylline alone at the above dosage. Lung function was evaluated prior to drug administration and then 3 and 7 days later. RESULTS: Oral administration of dexamethasone alone or combined with theophylline failed to improve lung function significantly in RAO affected horses. Theophylline alone also failed to improve lung function in all treated horses. Conversely, dexamethasone administration at 0.05 mg/kg bwt i.v. resulted in a significant improvement in lung function starting on Day 3. CONCLUSIONS AND POTENTIAL RELEVANCE: Oral theophylline for 7 days did not improve the effects of a low dose of dexamethasone for the treatment of horses with RAO. PMID- 17124850 TI - Physiotherapy applied to the horse: a review. PMID- 17124851 TI - Historical progression of racing performance in the Thoroughbred horse and man. PMID- 17124852 TI - [The times, they are a-changin']. PMID- 17124853 TI - [Tick-borne encephalitis, rabies, and echinococcosis in Switzerland]. AB - Swiss guidelines have been published recently for Lyme disease, but three other zoonoses have motivated a particular attention lately because they cause or might cause illnesses in Switzerland. The incidence of tick-borne encephalitis is clearly increasing since 2004. The vaccination of persons living or residing temporally in risk areas is now routinely recommended. Although eradicated in wild mammals since 1996, rabies still constitutes a threat through bats and illegally imported pets. The epidemiology of echinococcosis is influenced by the increase in fox populations in urban areas. This could result in a higher prevalence of the disease in dogs and thus a higher risk of exposure for humans. PMID- 17124854 TI - [Epidemiogy and treatment of tuberculosis in Switzerland]. AB - Problems in tuberculosis are its rarity, drug-resistant strains, and patients' social aspects. Knowing the epidemiology helps to "think tuberculosis" when some anamnestic and clinical findings are present. In a not too seriously ill patient, the emergency consists of excluding infectious tuberculosis by sputum smear examinations. A positive result will lead to the patient's isolation and the initiation of treatment. Negative smears allow to wait for culture results, while following the patient and repeating smears after a non-specific antibiotic treatment. In case of strong suspicion of tuberculosis or risk for a rapid progression, the treatment should begin without delay. Discussion with a specialist is often warranted. A follow-up by the Swiss Pulmonary Association is recommended to prevent the patient from defaulting from treatment. PMID- 17124855 TI - [Diagnosis and treatment of urethritis]. AB - Clinical urethritis should motivate urine analyses including tests for leukocytes and PCR for C. trachomatis and N. gononhocae. The culture of an urethral swab for gonococci is sensitive as well. It allows susceptibility tests. Other infectious agents have been associated with urethritis (e.g., M. genitalium, T. vaginalis, H. simplex). They should be searched in case of negative first results or non response to an empiric treatment. Given the emergence of resistances to quinolones, single doses of azithromycin 1 g plus cefixime 400 mg constitute a good choice which permits to ensure adherence. Contact tracing, screening for other sexually transmitted infections, and the evaluation of the sexual behaviour should always be done in order to prevent any further transmission. PMID- 17124856 TI - [The management of HIV infection: acute retroviral syndrome and follow-up principles]. AB - Current evidence no longer justifies the introduction of antiretroviral treatment during primary HIV infection. Consequently, patients presenting with the acute retroviral syndrome should not be treated outside clinical studies. For patients with chronic infection, the decline of opportunistic infections, due to the efficacy of highly active antiretroviral therapy, has left the place to daily problems of adverse effects and drug interactions. In Switzerland, the follow-up of HIV-infected adults is organized within the framework of a prospective cohort study. PMID- 17124857 TI - [Infections and TNFalpha antagonist treatment]. AB - The TNFalpha antagonists have become a standard treatment for many severe chronic inflammatory diseases. After a few years of practical use in individual patient's setting adverse effects, especially infectious have been recorded and a practical attitude can be proposed for prevention and follow-up. TNFalpha antagonists increase the risk of infection by intracellular pathogens. A latent tuberculosis infection must be looked for prior to the initiation of the treatment. A prophylaxis is started in case of positive result. The global infectious risk can also be minimized by an optimal vaccination coverage and a regular screening for leucopenia. PMID- 17124858 TI - [Autism and neuroscience]. AB - The understanding of autism has improved with neurosciences, especially with genetics, molecular biology and cerebral neuroimaging. This trouble looks much more heterogenous than expected, from a clinical and an etiological point of view as well. No clear pathopsychological model about the syndromes of the autistic spectrum can so far be proposed but several scientific breakthroughs seem interesting. PMID- 17124859 TI - [No one is expected to ignore... the legal status of drug promotional studies]. PMID- 17124860 TI - [A beautiful story between dependence and competition]. PMID- 17124861 TI - [New warnings on the pathologic vascular risks associated with NSAIDs]. PMID- 17124862 TI - [On the frontier of a new eugenics]. PMID- 17124863 TI - [The 2006 Nobel for RNA interference]. PMID- 17124864 TI - [Chronic otitis: the clamp squeezes around bacterial biofilms]. PMID- 17124865 TI - [On our back, our future?]. PMID- 17124866 TI - Molecular paleoscience: systems biology from the past. AB - Experimental paleomolecular biology, paleobiochemistry, and paleogenetics are closely related emerging fields that infer the sequences of ancient genes and proteins from now-extinct organisms, and then resurrect them for study in the laboratory. The goal of paleogenetics is to use information from natural history to solve the conundrum of modern genomics: How can we understand deeply the function of biomolecular structures uncovered and described by modern chemical biology? Reviewed here are the first 20 cases where biomolecular resurrections have been achieved. These show how paleogenetics can lead to an understanding of the function of biomolecules, analyze changing function, and put meaning to genomic sequences, all in ways that are not possible with traditional molecular biological studies. PMID- 17124867 TI - A structure-centric view of protein evolution, design, and adaptation. AB - Proteins, by virtue of their central role in most biological processes, represent one of the key subjects of the study of molecular evolution. Inherent in the indispensability of proteins for living cells is the fact that a given protein can adopt a specific three-dimensional shape that is specified solely by the protein's sequence of amino acids. Over the past several decades, structural biologists have demonstrated that the array of structures that proteins may adopt is quite astounding, and this has lead to a strong interest in understanding how protein structures change and evolve over time. In this review we consider a large body of recent work that attempts to illuminate this structure-centric picture of protein evolution. Much of this work has focused on the question of how completely new protein structures (i.e., new folds or topologies) are discovered by protein sequences as they evolve. Pursuant to this question of structural innovation has been a desire to describe and understand the observation that certain types of protein structures are far more abundant than others and how this uneven distribution of proteins implicates on the process through which new shapes are discovered. We consider a number of theoretical models that have been successful at explaining this heterogeneity in protein populations and discuss the increasing amount of evidence that indicates that the process of structural evolution involves the divergence of protein sequences and structures from one another. We also consider the topic of protein designability, which concerns itself with understanding how a protein's structure influences the number of sequences that can fold successfully into that structure. Understanding and quantifying the relationship between the physical feature of a structure and its designability has been a long-standing goal of the study of protein structure and evolution, and we discuss a number of recent advances that have yielded a promising answer to this question. Finally, we review the relatively new field of protein structural phylogeny, an area of study in which information about the distribution of protein structures among different organisms is used to reconstruct the evolutionary relationships between them. Taken together, the work that we review presents an increasingly coherent picture of how these unique polymers have evolved over the course of life on Earth. PMID- 17124868 TI - Mechanisms of protein evolution and their application to protein engineering. AB - Protein engineering holds great promise for the development of new biosensors, diagnostics, therapeutics, and agents for bioremediation. Despite some remarkable successes in experimental and computational protein design, engineered proteins rarely achieve the efficiency or specificity of natural enzymes. Current protein design methods utilize evolutionary concepts, including mutation, recombination, and selection, but the inability to fully recapitulate the success of natural evolution suggests that some evolutionary principles have not been fully exploited. One aspect of protein engineering that has received little attention is how to select the most promising proteins to serve as templates, or scaffolds, for engineering. Two evolutionary concepts that could provide a rational basis for template selection are the conservation of catalytic mechanisms and functional promiscuity. Knowledge of the catalytic motifs responsible for conserved aspects of catalysis in mechanistically diverse superfamilies could be used to identify promising templates for protein engineering. Second, protein evolution often proceeds through promiscuous intermediates, suggesting that templates which are naturally promiscuous for a target reaction could enhance protein engineering strategies. This review explores these ideas and alternative hypotheses concerning protein evolution and engineering. Future research will determine if application of these principles will lead to a protein engineering methodology governed by predictable rules for designing efficient, novel catalysts. PMID- 17124869 TI - Novel enzymes through design and evolution. AB - The generation of enzymes with new catalytic activities remains a major challenge. So far, several different strategies have been developed to tackle this problem, including site-directed mutagenesis, random mutagenesis (directed evolution), antibody catalysis, computational redesign, and de novo methods. Using these techniques, a broad array of novel enzymes has been created (aldolases, decarboxylases, dehydratases, isomerases, oxidases, reductases, and others), although their low efficiencies (10 to 100 M(-1) s(-l)) compared to those of the best natural enzymes (10(6) to 10(8) M(-1) s(-1)) remains a significant concern. Whereas rational design might be the most promising and versatile approach to generating new activities, directed evolution seems to be the best way to optimize the catalytic properties of novel enzymes. Indeed, impressive successes in enzyme engineering have resulted from a combination of rational and random design. PMID- 17124871 TI - Nurse practitioners in the ACT. PMID- 17124872 TI - Making a difference where it counts. Interview by Philip Luker. PMID- 17124873 TI - Pressure rising against violence. PMID- 17124874 TI - Nursing and the 5As guideline to smoking cessation interventions. PMID- 17124875 TI - Compassion with dignity and respect. PMID- 17124876 TI - Have you seen the 10-year long-term safety data on LASIK? PMID- 17124877 TI - Intraocular lens power calculation for eyes with previous myopic LASIK. PMID- 17124878 TI - Prediction and control of corneal asphericity after refractive surgery. PMID- 17124879 TI - Probability model of the inaccuracy of residual stromal thickness prediction to reduce the risk of ectasia after LASIK part I: quantifying individual risk. AB - PURPOSE: To measure the imprecision of microkeratome cuts, preoperative corneal pachymetry, and laser ablation depth and develop a statistical model to describe the probability of the residual stromal bed thickness (RST) after myopic LASIK being significantly thinner than predicted. METHODS: Preoperative corneal thickness, flap thickness, ablation depth, and RST were measured in 36 eyes by a prototype three-dimensional very high-frequency (VHF) 50 MHz digital ultrasound scanning device (<1.2 microm precision), precursor to the commercially available Artemis 2. All eyes had undergone LASIK with the Moria LSK-One microkeratome and the NIDEK EC-5000 excimer laser. Based on the statistically combined uncertainty (standard deviation) and bias (accuracy to intended value) of corneal thickness measurement, flap thickness, and ablation depth, a continuous probability function was devised describing the chance of obtaining an actual RST less than a specified "cut-off". The model was applied using the data collected from the cohort of eyes. The model was also applied using published flap thickness statistics on a series of microkeratomes. RESULTS: Precision (standard deviation) was 0.74 microm for VHF digital ultrasound measurement of pachymetry, 30.3 microm for Moria LSK-One flap thickness, and 11.2 microm for NIDEK EC-5000 ablation depth. Assuming negligible laser ablation depth bias, the model found the probability that the actual RST will be <200 pmicromgiven a target RST of 250 microm is 7.56% with the Moria LSK-One. The model applied to published flap statistics revealed a range of probabilities of leaving <200 microm given a target RST of 250 microm from <0.01% to 33.6%. CONCLUSIONS: The choice of microkeratome, laser, and pachymeter has a significant impact on the variation of the depth of keratectomy and thus on the risk of ectasia. This model together with high-precision microkeratomes, preoperative pachymetry, and knowledge of laser ablation precision would enable surgeons to determine the specific imprecision of RST prediction for individual LASIK cases and minimize the risk of ectasia. PMID- 17124880 TI - Probability model of the inaccuracy of residual stromal thickness prediction to reduce the risk of ectasia after LASIK part II: quantifying population risk. AB - PURPOSE: To derive a statistical model to estimate the rate of excessive keratectomy depth below a selected cut-off residual stromal thickness (RST) given a minimum target RST and specific Clinical Protocol; apply the model to estimate the RST below which ectasia appears likely to occur and back-calculate the safe minimum target RST that should be used given a specific Clinical Protocol. METHODS: Myopia and corneal thickness distribution were modeled for a population of 5212 eyes that underwent LASIK. The probability distribution of predicted target RST error (Part I) was used to calculate the rate of excessive keratectomy depth for this series. All treatments were performed using the same Clinical Protocol; one surgeon, Moria LSK-One microkeratome, NIDEK EC-5000 excimer laser, Orbscan pachymetry, and a minimum target RST of 250 microm--the Vancouver Clinical Protocol. The model estimated the RST below which ectasia appears likely to occur and back-calculated the safe minimum target RST. These values were recalculated for a series of microkeratomes using published flap thickness statistics as well as for the Clinical Protocol of one of the authors-the London Clinical Protocol. RESULTS: In the series of 5212 eyes, 6 (0.12%) cases of ectasia occurred. The model predicted an RST of 191 microm for ectasia to occur and that a minimum target RST of 329 microm would have reduced the -rate of ectasia to 1: 1,000,000 for the Vancouver Clinical Protocol. The model predicted that the choice of microkeratome varied the rate of ectasia between 0.01 and 11,623 eyes per million and the safe minimum target RST between 220 and 361 microm. The model predicted the rate of ectasia would have been 0.000003: 1,000,000 had the London Clinical Protocol been used for the Vancouver case series. CONCLUSIONS: There appears to be no universally safe minimum target RST to assess suitability for LASIK largely due to the disparity in accuracy and reproducibility of microkeratome flap thickness. This model may be used as a tool to evaluate the risk of ectasia due to excessive keratectomy depth and help determine the minimum target RST given a particular Clinical Protocol. PMID- 17124881 TI - Twelve-year follow-up of photorefractive keratectomy for low to moderate myopia. AB - PURPOSE: To evaluate long-term safety and stability in a group of myopic patients who underwent photorefractive keratectomy (PRK) > or =12 years ago. METHODS: Myopic PRK was performed on 120 eyes of 80 patients using the Summit UV200 excimer laser with a 5-mm ablation zone. Of the original group, most of whom were followed for > or =2 years (mean 2.6 +/- 1.7 years), 34 patients (58 eyes) returned at 12 years (mean 12.7 +/- 0.79 years) and had refractive stability, refractive predictability, best spectacle-corrected visual acuity (BSCVA), corneal haze, and subjective patient symptoms, such as glare/halos, recorded. RESULTS: Preoperative mean refractive spherical equivalent (MRSE) ranged from 1.75 to -7.25 diopters (D) and astigmatism from 0.00 to 1.50 D. All eyes underwent a change in manifest refraction over 12 years. At 2 years, MRSE was 0.27 +/- 0.55 D and at 12 years was -0.58 +/- 0.72 D. In 87.9% of eyes, the level of preoperative BSCVA was maintained or improved, whereas 34.5% of eyes gained one line, and 12.1% lost one line of BSCVA. Uncorrected visual acuity > or = 20/20 was noted in 67% of eyes, whereas 62.1% were within +/- 0.50 D of emmetropia. Trace haze was noted in 17.2% of eyes at 12 years. One patient had a rhegmatogenous retinal detachment, but this was unlikely due to the PRK procedure. With respect to the small optical zone, 14 (41.1%) patients had night visual problems, particularly halos, which were severe in 2.7%. All patients questioned stated they would have the procedure done again. CONCLUSIONS: Photorefractive keratectomy with the Summit UV200 excimer laser effectively reduced myopia and showed good refractive stability from year 2 to 12 with good patient satisfaction. PMID- 17124882 TI - Ten-year follow-up of 360 degrees intrastromal corneal rings for myopia. AB - PURPOSE: To determine the safety, efficacy, and stability of intrastromal corneal rings 10 years after placement for myopia. METHODS: Ten eyes with myopia treated with the placement of 360 degrees complete intrastromal corneal rings with the ends sutured together were evaluated at 10-year follow-up for the following: uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), manifest refraction spherical equivalent (MRSE), induced manifest refractive cylinder, keratometry readings, slit-lamp findings, and ultrasound central pachymetry. Ten-year data were compared to 1-year results to assess visual stability over time. RESULTS: No statistically significant difference was noted between UCVA at 1 year and 10 years. At the 10-year examination, 90% of patients had BSCVA > or = 20/25 and 100% had BSCVA > or = 20/30. There was no statistically significant difference between MRSE at 1 year and 10 years. At the 10-year examination, induced manifest refractive cylinder was >1.00 diopter (D) in 0% of eyes and < or = 0.25 D in 60% of eyes. There was no statistically significant difference between mean central keratometric power at 1-year follow up compared with 10-year follow-up. No statistically significant difference was noted in central corneal thickness between 1 and 10 years in the eyes studied. CONCLUSIONS: Intrastromal corneal rings are an effective and stable method of correcting mild myopia based on optic parameters. PMID- 17124883 TI - Traumatic corneal flap dislocation one to six years after LASIK in nine eyes with a favorable outcome. AB - PURPOSE: To report our experience treating eye trauma after LASIK refractive surgery. METHODS: Nine eyes of eight patients (one woman and seven men) were treated for ocular trauma: blunt trauma (n=5), sharp instrument trauma (n=2,) and trauma from inflation of automobile air bags during a traffic accident (n=2). The time from LASIK varied between 3 months and 6 years. All patients were hospitalized as a result of severe decrease in visual acuity and pain. RESULTS: Seven of nine LASIK flaps had some degree of dislocation and were lifted, irrigated, and repositioned. Two flaps were edematous without dislocation. Intensive topical steroids and antibiotics were used in all patients up to 3 weeks after trauma. Three months after trauma, five eyes regained their pre trauma visual acuity (between 20/20 and 20/40), and three eyes lost one line of best spectacle-corrected visual acuity. CONCLUSIONS: Trauma occurring several months or years after LASIK may cause flap injury. Adequate and prompt treatment usually is successful. PMID- 17124884 TI - Clinical results with the medennium phakic refractive lens for the correction of high myopia. AB - PURPOSE: To evaluate the predictability, safety, stability, complications, and biocompatibility of the phakic refractive lens (PRL) as a posterior chamber intraocular lens to correct high myopia. METHODS: Fifty eyes of 31 patients who underwent posterior chamber PRL implantation were evaluated prospectively. Mean preoperative myopia was -12.54 +/- 4.22 diopters (D) (range: -4.50 to -23.50 D) and mean astigmatic refractive power was -1.38 +/- 1.24 D (range: -1.00 to -4.50 D). Surgical implantation was performed through a 3.0- to 4.0-mm clear cornea sutureless incision using parabulbar (sub-Tenon's) anesthesia. Intra- and postoperative complications were recorded. RESULTS: Three months after surgery, the mean spherical equivalent refraction was -0.21 +/- 0.42 D (range: +1.00 to 1.75 D). At 6 and 12 months, mean spherical equivalent refraction was -0.23 +/- 0.38 D (range: 0 to -1.25 D). At the last examination, uncorrected visual acuity was > or = 20/40 in 41 (82%) eyes and > or = 20/20 in 22 (44%) eyes. Best spectacle-corrected visual acuity (BSCVA) was > or = 20/40 in 42 (84%) eyes and > or = 20/20 in 27 (54%) eyes. Comparison of pre- and postoperative BSCVA at 12 months showed that 12 (36.4%) of 33 eyes gained > or =1 lines of BSCVA and 7 (21.2%) of 33 eyes gained > or =2 lines. One (2%) eye developed anterior subcapsular cataract requiring lens exchange, and 1 (2%) eye developed acute angle closure glaucoma requiring YAG-iridotomy. One (2%) eye developed macular hemorrhage. CONCLUSIONS: At 6 months and 1 and 2 years, PRL implantation yielded encouraging visual and refractive results with excellent biocompatibility. The efficacy, stability, and short-term safety of this lens was established. Serious complications, such as cataract and acute angle closure glaucoma, may occur, and long-term safety needs to be evaluated. PMID- 17124885 TI - Higher order aberrations in normal eyes measured with three different aberrometers. AB - PURPOSE: To assess the repeatability of measurements of higher order aberrations using three different aberrometers and to compare higher order aberration measurements between optical path difference (OPD) scanning and the Hartmann Shack method. METHODS: Wavefront aberration data obtained using the NIDEK OPD Scan, Bausch & Lomb Zywave wavefront aberrometer, and VISX CustomVue wavefront analyzer were compared. A total of 19 subjects were included in the study. The repeatability in each machine was assessed by calculating the difference between measurements and the mean of three consecutive measurements in the same eye. Subsequent analysis of the distribution of these differences yields the mean difference, the standard deviation of the differences, and the 95% confidence interval for repeated measurements, also termed the "repeatability coefficient". RESULTS: Repeatability errors in all three machines were found to be low, suggesting that all three machines are reliable in their repeated measurements. Significant differences were demonstrated between OPD scanning and Hartmann-Shack aberrometers. All three machines showed statistically significant differences in several higher order aberration parameters when compared to each other. CONCLUSIONS: The three different aberrometers provided repeatable measurements but statistical differences were noted in the measurement of higher order aberrations when comparing the machines. No instrument was superior over the other and all three were reliable. PMID- 17124886 TI - Laceration and partial dislocation of LASIK flaps 7 and 4 years postoperatively with 20/20 visual acuity after repair. AB - PURPOSE: To report two cases of late ocular trauma resulting in laceration of corneal flaps 7 and 4 years (cases 1 and 2, respectively) after LASIK. METHODS: A 49-year-old man sustained laceration of the flap and interface foreign body debris from construction material in the right eye 7 years after uneventful LASIK (case 1). A 33-year-old man had partial dislocation of the flap 4 years after LASIK in the left eye (case 2) due to a penetrating wood chip accident. Surgical procedures were performed to remove the foreign bodies and reposition the flaps. RESULTS: In case 1, postoperative uncorrected visual acuity (UCVA) was 20/25 with manifest refraction of -0.50 -0.50 x 110 corrected to 20/20. In case 2, UCVA was 20/20 with piano refraction after LASIK retreatment. CONCLUSIONS: Corneal LASIK flaps are susceptible to penetrating trauma, which can occur >7 years after the procedure. Proper everamanage ment of the trauma and LASIK flap can restore excellent visual acuity. PMID- 17124887 TI - ZB5M anterior chamber and Fyodorov's posterior chamber phakic intraocular lenses: long-term follow-up. AB - PURPOSE: To compare the long-term complications of two outdated phakic intraocular lenses (PIOL) implanted in eyes with high myopia to aid development of better PIOL designs. METHODS: Sixty-two eyes of 37 patients implanted with the ZB5M anterior chamber PIOL (anterior chamber group) and 22 eyes of 14 patients implanted with Fyodorov's posterior chamber PIOL (posterior chamber group) were examined. Complications were compared after a mean follow-up of 71.15 +/- 32.17 months (range: 24 to 145 months) in the anterior chamber group and 84.14 +/- 26.43 months (range: 24 to 132 months) in the posterior chamber group (P = .07). RESULTS: At last follow-up, pupil ovalization was present in 32 (51.6%) eyes in the anterior chamber group and 1 (4.5%) eye in the posterior chamber group (P < .001), and cataract in 5 (8.1%) eyes in the anterior chamber group and 7 (31.8%) eyes in the posterior chamber group (P = .007). Phakic intraocular lens decentration, glaucoma, retinal detachment, corneal decomposition, and monocular diplopia were additional complications encountered in both groups, with no statistically significant difference. CONCLUSIONS: Proper understanding of these outdated PIOLs may be helpful in developing better PIOL designs. PMID- 17124888 TI - Bilateral central crystalline corneal deposits four years after intacs for myopia. AB - PURPOSE: To report a case of bilateral central crystalline keratopathy in the anterior stroma occurring 4 years after Intacs implantation. METHODS: A 45-year old woman underwent bilateral uncomplicated Intacs implantation for myopia. The postoperative course was uneventful. However, between 3 and 4 years after surgery, the patient developed central opacifications of the anterior stroma in both eyes, reducing best spectacle-corrected visual acuity. RESULTS: Intacs were explanted. Confocal microscopy, electron microscopy of the explanted ring segments, and microbiology studies were performed. Opacities were still detectable at the slit-lamp microscope up to 8 months after explantation. CONCLUSIONS: This is the first report on central corneal opacifications after Intacs implantation for myopia. The opacities could be the result of chronic metabolic stress or the beginning of lipid-like changes in another more central corneal localization. PMID- 17124889 TI - Reliability of corneal and total wavefront aberration measurements with the SCHWIND Corneal and Ocular Wavefront Analyzers. AB - PURPOSE: To evaluate the reliability of the SCHWIND Corneal and Ocular Wavefront Analyzers. METHODS: This study comprised 115 eyes of 58 healthy volunteers (26 [44.8%] women and 32 [55.2%] men) with no corneal or lenticular pathologies and normal visual acuity. All eyes underwent three consecutive measurements by one examiner with the Corneal Wavefront Analyzer and the Ocular Wavefront Analyzer (Hartmann-Shack principle). Corneal wavefront errors were calculated using the topography data, a standard eye model, and ray tracing. The reliability was tested for 2nd, 3rd, and 4th order aberrations as mean values of the standard deviations for all measurements. RESULTS: Mean patient age was 23 +/- 2.1 years. The mean refraction was -0.77 +/- 1.56 diopters (D) (range: +3.33 to -5.28 D). The repeatability test revealed a good reliability for both machines with a slightly better value for the Ocular Wavefront Analyzer for 3rd and 4th order higher order aberrations (P < .05, Wilcoxon test). CONCLUSIONS: Wavefront measurements of corneal and total optical aberrations performed with the Corneal Wavefront Analyzer and the Ocular Wavefront Analyzer have good reliability. Both measurements are recommended prior to any refractive surgery treatment. PMID- 17124890 TI - Corneal first surface wavefront aberrations before and after pterygium surgery. AB - PURPOSE: To determine the higher order aberrations at the corneal first surface before and after surgery for pterygium. METHODS: Data were drawn from a longitudinal study of patients undergoing pterygium excision at Royal Victoria Infirmary, Newcastle upon Tyne, England between September 1998 and May 2004. Corneal topography was taken with the TMS-2 Topographic Modeling System (Computed Anatomy Corp) prior to and 6 months after surgery, exported to VOLPro software v7.08 (Sarver & Associates), and wavefront aberrations were derived for a 5.0-mm pupil using a 10th order Zernike polynomial expansion. Pre- to postoperative changes were assessed for significance using analyses of variance, and the relative risk of significant postoperative aberrations by pterygium size was determined. RESULTS: Satisfactory corneal topography was available on 67 eyes (mean age 53.8 +/- 16.7 years [range: 25-86 years]). The root-mean-square (RMS) fit error in preoperative eyes was 0.15 +/- 0.10 microm. Preoperatively, the total higher order RMS wavefront aberration was 0.94 +/- 0.83 microm. All Zernike modes were elevated, with trefoil being the major contributor 0.52 +/- 0.50 microm. Pterygium excision significantly reduced wavefront aberrations across all modes and orders (F(1, 129) = 6.7 to 22.6, P < .01): total higher order RMSpostop 0.45 +/- 0.35 microm. Cases with visually significant postoperative aberrations occurred and were more likely with larger pterygia: relative risk compared to pterygia 1.0 to 1.9 mm was 1.3 for 2.0 to 2.9 mm, 8.5 for 3.0 to 3.9 mm, 13.3 for 4.0 to 4.9 mm, and 10.2 for 5.0 to 5.9 mm. CONCLUSIONS: Zernike polynomial fitting well describes wavefront aberrations in eyes with pterygia. Pterygia are associated with wavefront aberrations, especially trefoil, but these were largely eliminated by surgery. Earlier excision of pterygia reduces the likelihood of significant residual aberrations. PMID- 17124891 TI - Penetrating keratoplasty for keratoconus: the nexus between corneal wavefront aberrations and visual performance. AB - PURPOSE: To compare the visual and optical performance after penetrating keratoplasty (PK) for keratoconus to normal patients and to examine the relationship between corneal wavefront aberrations and visual performance in patients with PK. METHODS: Visual performance testing, with optimal refractive correction, included low contrast visual acuity (LCVA) and Pelli-Robson contrast sensitivity with and without glare, and high contrast visual acuity. Corneal first surface wavefront aberrations were calculated from EyeSys topography data using VOL-Pro software v7.00 for a 4.0-mm pupil as a 10th order Zernike expansion and converted into single value metrics. Normal patients were compared to patients with PK using analysis of variance, and linear regression was used to compare wavefront aberration metrics to visual performance. RESULTS: Patients with PK (n=14, age 41.6 +/- 7.0 years) and normal patients (n=14, age 36.7-9.0 years) were of similar age (F(1, 26) = 2.54, P = .12). Normal patients saw significantly better on all visual performance measures and had better optical performance for total higher order root-mean-square corneal wavefront aberration (mean-SD): PK, 0.67 +/- 0.41 microm; normal, 0.09 +/- 0.02 microm (F(1,26) = 28.41, P < .001) and across all Zernike orders and modes. Wavefront aberrations in PK eyes were dominated by trefoil 0.35 +/- 0.27 microm, coma 0.47 +/- 0.37 microm, spherical aberration 0.17 +/- 0.10 microm, and tetrafoil 0.12 +/- 0.07 microm. The relationships between corneal wavefront aberration and visual performance metrics were strongest for LCVA = 0.30-0.98 Pupil fraction for wavefront (tessellation) -0.04 Half width at half height, R2=0.75. CONCLUSIONS: In this series, patients with PK had poorer visual performance compared to normal patients, which is due to increased corneal wavefront aberrations. Outcomes research in corneal transplantation should include measurement of wavefront aberrations and visual performance in the contrast domain. PMID- 17124892 TI - A Shack-Hartmann-based autorefractor. AB - PURPOSE: Autorefractors are typically based on either the optometer or the Scheiner principles, or a combination of the two techniques. These devices have dominated the market for objective assessment of refractive error for >30 years. The purpose of this investigation is to test a Shack-Hartmann-based system as an alternative to these systems. METHODS: Fourteen subjects with varying levels of refractive error were measured with a Topcon autorefractor and a Shack-Hartmann based autorefractor. Fourier transform techniques were used to extract sphere, cylinder, and axis information from the Shack-Hartmann images, avoiding the need for image processing. The deviation of the refractive error from a subjective refraction was used as a means of comparing the two devices. RESULTS: The two devices performed similarly on this group of subjects. The mean difference in refraction between the two devices was nearly zero, suggesting that the likelihood and magnitude of errors for the two devices are equivalent. CONCLUSIONS: The Shack-Hartmann-based autorefractor shows promise as an alternative to conventional optometer or Scheiner-based technologies. However, issues with extending the myopic range of the device still need to be resolved. PMID- 17124893 TI - Shack-Hartmann-Based wavefront analyzer for calibrating excimer lasers. AB - PURPOSE: Currently, refractive surgical excimer laser systems are calibrated by ablating plastic lenses, which are measured by lensometer and analyzed by a technician. The accuracy of this method is approximately 0.25 diopters (D) in sphere and cylinder power. Theoretically, objective calibration using wavefront technology would be significantly more accurate, thereby improving surgical outcomes. This study describes a Shack-Hartmann-based instrument, which has been developed to measure ablated plastic lenses for calibration and quality control of the excimer laser. METHODS: A calibration instrument comprising an LED source at 640 nm, a lenslet array, beam-guiding optics, and a CCD camera was designed to perform full wavefront analysis. The measurement plane is conjugate to the lenslet array plane, and the diameter of the pupil is 5 mm. Accuracy was determined by measuring a set of well-calibrated spherical and cylindrical glass lenses. Plastic lenses were ablated, and high-precision measurements were performed by surface profile scanner. RESULTS: In the power range of -6.00 to +4.00 D, repeatability exceeded 0.01 D, accuracy of measurement exceeded 0.04 D, and 10 for the axis of cylinder lenses. The measurement of excimer-ablated plastic lenses agreed with high-precision surface profile scanner measurements within 0.10 D, and repeatability exceeded 0.01 D. CONCLUSIONS: Wavefront technology-based, high-precision measurement of calibration lenses can more accurately set the energy of the excimer laser, which enhances the accuracy of refractive laser correction. In automating calibration, the new instrument removes operator subjectivity and decreases the time needed for calibration. PMID- 17124894 TI - Comparison of wavefront reconstructions with Zernike polynomials and Fourier transforms. AB - PURPOSE: To make a direct comparison between Fourier and Zernike reconstructions of ocular wavefronts using a newly available analytical theory by which Fourier coefficients can be converted to Zernike coefficients and vice versa. METHODS: Noise-free random wavefronts were simulated with up to the 15th order of Zernike polynomials. For each case, 100 random wavefronts were simulated separately. These wavefronts were smoothed with a low-pass Gaussian filter to remove edge effects. Wavefront slopes were calculated, and normally distributed random noise was added within the circular area to simulate realistic Shack-Hartmann spot patterns. Three wavefront reconstruction methods were performed. The wavefront surface error was calculated as the percentage of the input wavefront root mean square. RESULTS: Fourier full reconstruction was more accurate than Zernike reconstruction from the 6th to the 10th orders for low-to-moderate noise levels. Fourier reconstruction was found to be approximately 100 times faster than Zernike reconstruction. Fourier reconstruction always makes optimal use of information. For Zernike reconstruction, however, the optimal number of orders must be chosen manually. The optimal Zernike order for Zernike reconstruction is lower for smaller pupils than larger pupils. CONCLUSIONS: Fourier full reconstruction is faster and more accurate than Zernike reconstruction, makes optimal use of slope information, and better represents ocular aberrations of highly aberrated eyes. PMID- 17124895 TI - Extracting wavefront error from Shack-Hartmann images using spatial demodulation. AB - PURPOSE: To determine whether the spatial demodulation processing of Shack Hartmann images is suitable for extracting wavefront gradients for ocular wavefront sensors. METHODS: We developed a custom software program to implement the spatial demodulation technique. To test the algorithm's performance, we generated simulated spot images and obtained an eye examination image. We generated a collection of simulated aberrated spot images corresponding to: astigmatic wavefront (-5.00 -2.00 x 17), highly aberrated defocus (+/-20.00 diopters [D]), high-resolution defocus (-0.01 D), and third-order aberrations (trefoil and coma). The eye examination image and its measured Zernike coefficients were obtained from a Shack-Hartmann ocular aberrations system. We evaluated the output from the algorithm in terms of comparing the results to the known Zernike coefficients (for the simulated images) or the previously measured Zernike coefficients (for the eye examination image). RESULTS: The spatial demodulation algorithm was able to correctly recover the aberrations to better than 1/100 (0.01) D for the simulated spot images. The processing of the eye examination image yielded results within approximately 1/4 (0.25) D to the values provided by the Shack-Hartmann system. CONCLUSIONS: From the set of simulated images and the eye examination image used to test the spatial demodulation technique, it appears that the method is suitable for application in ocular wavefront aberrations Shack-Hartmann systems. The method appears capable of accurately processing high levels of aberrations (+/-20.00 D) as well as providing high resolution as evidenced by finding the -0.01 D defocus. The method may be especially well suited for processing highly aberrated wavefronts. PMID- 17124896 TI - A new wavefront sensor with polar symmetry: quantitative comparisons with a Shack Hartmann wavefront sensor. AB - PURPOSE: A novel wavefront sensor has been developed. It follows the same principle of the Shack-Hartmann wavefront sensor in that it is based on slope information. However, it has a different symmetry, which may offer benefits in terms of application. METHODS: The new wavefront sensor consists of a set of donut-shaped acrylic lenses with a charge coupled device located at the focal plane. From detection of shift in the radial direction, radial slopes are computed for 2880 points. Theoretical computations for higher order aberrations and lower order aberrations were implemented for the Shack-Hartmann wavefront sensor and the new wavefront sensor, and practical measurements were conducted on several sphere-cylinder trial lenses. RESULTS: The overall mean value of root mean square error (RMSE) (in microns) for theoretical computations was 0.03 for the Shack-Hartmann wavefront sensor and 0.02 for the new wavefront sensor. The mean value of RMSE for lower order aberrations (1-5) was 0.01 and 0.00003, and for higher order aberrations was 0.02 and 0.02, for the Shack-Hartmann and new wavefront sensors, respectively. For practical measurements (sphere, cylinder, axis), the standard deviation was 0.04 diopters (D), 0.04 D, and 4 degrees for the new wavefront sensor and 0.02 D, 0.02 D, and 5 degrees for the Shack-Hartmann wavefront sensor. CONCLUSIONS: Precision of the new wavefront sensor when measuring astigmatic and spherical surfaces is compatible with the Shack-Hartmann wavefront sensor. Centration with this new sensor is an absolute process using the center of the entrance pupil, which is where the line of site passes. This wavefront sensor, similar to the Shack-Hartmann sensor, does not eliminate the possibility of tilt. For more conclusive and statistically valid data, in vivo measurements are needed. PMID- 17124897 TI - Higher order aberrations comparison in fellow eyes following intraLase LASIK with wavelight allegretto and customcornea LADArvision4000 systems. AB - PURPOSE: To compare the differences in objective and subjective visual outcomes with wavefront-optimized versus wavefront-guided excimer laser ablations using two different excimer laser platforms. METHODS: In this prospective, randomized, matched, contralateral eye study, one eye was treated with the Alcon CustomCornea system, and the fellow eye was treated with the WaveLight Allegretto Wavefront Optimized system. The IntraLase FS30 femtosecond laser was used to create all flaps. Treatments were randomized to the dominant eye, and optical treatment zones were matched to 6.5 mm. Patients were observed for 3 months after surgery. Initial data analysis was available for the first 28 eyes of 14 patients. RESULTS: The only statistically significant differences between the groups were seen in the amount of higher order aberrations-the 14 eyes in the WaveLight group had a mean preoperative root-mean-square (RMS) error of 0.37 microm compared to 0.36 microm for 14 eyes in the Alcon group. Of the 8 patients who had 3-month follow-up, 8 eyes in the WaveLight group had a mean RMS error of 0.58 microm at 3 months, compared to 0.41 microm in the 8 eyes in the Alcon group. CONCLUSIONS: Early results from this study have shown that both laser platforms produce accurate refractive outcomes; however, the Alcon CustomCornea LADARVision4000 system induces statistically significantly fewer higher order aberrations than the WaveLight Allegretto system. PMID- 17124898 TI - Fulfilling the promise of laser refractive surgery. AB - PURPOSE: To review the characteristics required to perform accurate and precise laser refractive surgery relative to the current state of the art and the technology improvements needed in the future. METHODS: The general characteristics for laser refractive surgery are identified and their relative contributions to a successful result considered. For each characteristic, the degree to which improvements in technology are likely to improve results is discussed. RESULTS: The environment (surgeon, site), technology (laser, aberrometer), and patient are identified as key factors affecting results. Although aberrometers appear to be clinically sufficient, improvements in laser beam characteristics and delivery should improve system consistency. Current registration technology may be a factor in variability from patient to patient, but the greatest contribution in this regard would appear to be patient biomechanical and healing response factors. CONCLUSIONS: Wavefront-guided laser refractive surgery has demonstrated an ability to be better than conventional surgery. Improvements in laser technology, registration technology, and biomechanical diagnostics are expected to further improve results. These potential improvements are expected to reduce variability and suggest that the final potential of this technology has not yet been realized. PMID- 17124899 TI - The ideal excimer beam for refractive surgery. AB - PURPOSE: To identify essential considerations for the optimal excimer laser beam for refractive laser surgery and further to determine specific beam parameters fulfilling the essential requirements. METHODS: Starting with the fundamental laser tissue interaction, the various parameters defining the laser beam are analyzed and optimal values for each are evaluated. RESULTS: Gaussian beams generally produce smoother ablated surfaces than "flat top" beams. Optimal ablation efficiency for a Gaussian profile occurs at a peak laser fluence of approximately 450 mJ/cm2. To treat realistic optical aberrations the Gaussian spot size should be < 0.9 mm. The laser repetition rate for such a beam can be multiple hundred hertz without causing thermal damage if a "flying spot" strategy is used. Accurate treatment under such conditions requires both high-fidelity laser spot placement and high-performance eye tracking. CONCLUSIONS: Surgical systems using the characteristics defined in this study should yield excellent treatment outcomes. PMID- 17124900 TI - Reproducibility of wavefront measurements using the LADARWave aberrometer. AB - PURPOSE: To test reproducibility of lower and higher order aberration measurements using the Alcon LADAR-Wave system. METHODS: Measurements were obtained using the LADARWave aberrometer from 100 eyes (50 patients) who presented for refractive surgery. Each patient underwent 2 separate examinations a minimum of 1 day apart. Zernike polynomial values for these 2 separate measurements were recorded based on a calculated pupil size of 6.5 mm; reproducibility was assessed by analyzing the mean difference between the 2 measurements. RESULTS: The mean difference for defocus was 0.17 +/- 0.12 microm, which represented 2.6% +/- 1.9% of the mean defocus value (6.33 microm). The mean difference for astigmatism was 0.11 +/- 0.12 microm, which represented 23% +/- 26% of the mean astigmatism value (0.45 microm). The mean difference for coma was 0.06 +/- 0.06 microm, which represented 37% +/- 37% of the mean coma value (0.16 microm). The mean difference for spherical aberration was 0.04 +/- 0.04 microm, which represented 18% +/- 18% of the mean spherical aberration value (0.22 microm). CONCLUSIONS: Measurements for defocus were reproducible whereas measurements of astigmatism and higher order aberrations were more variable. The absolute root-mean-square differences between measurements were small, despite the many possible sources of error and variability. Surgical outcomes suggest that the LADARWave aberrometer is measuring at an appropriate clinical resolution. PMID- 17124901 TI - The LADAR6000: results in highly myopic and highly astigmatic eyes. AB - PURPOSE: To investigate the early clinical outcomes of the Alcon LADAR6000 system in treating two difficult patient populations: those with high myopia and those with high astigmatism. METHODS: Two series of eyes were of interest and subsequently enrolled in the study. Thirty-three eyes with preoperative manifest refraction spherical equivalent (MRSE) > -6.00 diopters (D) (high myopia group) and 25 eyes with preoperative myopic astigmatism > -1.50 D (high myopic astigmatism group) were enrolled. Mean patient age was 35 years in the high myopia group and 33 years in the astigmatism group. For the high myopia group, the mean preoperative MRSE was -8.13 D (range: -6.25 to -12.5 D). For the high myopic astigmatism group, the mean preoperative astigmatism was -1.89 D (range: 1.50 to -4.50 D). All LASIK flaps were made using a femtosecond laser with superior hinge position. Stromal ablation was performed using the LADAR6000 system for conventional treatment. All eyes were targeted for emmetropia. RESULTS: Both groups of eyes experienced excellent clinical outcomes with 100% of eyes within +/-1.00 D of attempted correction at 1 month. For the high myopia group, mean postoperative MRSE was -0.10 D at 1 month. For the high myopic astigmatism group, mean postoperative MRSE was -0.06 D at 1 month. CONCLUSIONS: The LADAR6000 system produced excellent early clinical outcomes in two challenging patient groups using LASIK with femtosecond laser flaps. PMID- 17124902 TI - Wavefront-guided treatment of symptomatic eyes using the LADAR6000 excimer laser. AB - PURPOSE: To evaluate the clinical results of wavefront-guided customized treatment of eyes that underwent previous keratorefractive surgery with clinically significant visual symptoms related to the presence of higher order aberrations, with or without defocus/astigmatism. METHODS: Twenty-eight consecutive eyes of 19 symptomatic patients underwent wavefront-guided custom photorefractive keratectomy (PRK with mitomycin C) to correct low and high order aberrations, using the LADAR-Vision platform. All eyes had manifest refractive spherical equivalent (MRSE) error from 0.00 to -4.25 diopters (D), and high order aberrations root-mean-square (RMS) values >0.80 microm for a 6.0-mm measurement diameter (mean: 1.18 microm). Eyes were examined at 1, 3, and 6 months postoperatively. Uncorrected (UCVA) and best spectacle-corrected visual acuity (BSCVA), MRSE, wavefront measurement of high order aberrations, and patient subjective evaluation were the parameters used to assess the treatment. RESULTS: Six months after treatment, all 28 eyes had an improvement in UCVA compared to preoperative values. Seventeen (60.7%) eyes gained lines of BSCVA (1 to 3 lines) whereas 1 eye lost 1 line. Although the amount of positive defocus was intentionally and empirically undercorrected in this study, MRSE was slightly hyperopic (mean +0.65 D, range: -0.25 to +1.75 D). All eyes showed a reduction in pre-existing high order aberrations, with a mean reduction of 47% (0.62 microm). Spherical aberration reduced the most (mean 60%), coma was reduced by a mean 29%, and all other terms were reduced by a mean 42% compared to preoperative values. Subjectively, all patients but 1 (both eyes treated) noticed a reduction in their pre-existing visual symptoms. Overall, the treatment was considered satisfactory by the patients in terms of visual quality gain. CONCLUSIONS: This series of consecutive treatments of symptomatic eyes indicates wavefront-guided custom ablation is an effective surgical option to reduce visually impairing high order aberrations and related visual symptoms. To avoid the resulting hyperopic shift, some significant nomogram adjustments are necessary, most often when pre-existing positive spherical aberration is present. PMID- 17124903 TI - Changing times. PMID- 17124904 TI - Obedience to the unenforceable. PMID- 17124905 TI - Balancing paradoxes. PMID- 17124907 TI - New CMS1500 claim form. PMID- 17124906 TI - Bird flu overview "best coverage" of state guard activities. PMID- 17124908 TI - Concierge medicine ... another option for frustrated doctors? PMID- 17124909 TI - Failure to communicate. PMID- 17124910 TI - Heroes. PMID- 17124911 TI - Is there life left in non-competes?--Part I. Physician non-compete agreements in contract negotiations. PMID- 17124912 TI - Choosing an executor. PMID- 17124913 TI - Blood loss in the space of Retzius and pelvis with tension-free vaginal tape and trans-obturator tape procedures. AB - OBJECTIVES: The purpose of this study was to evaluate the space of Retzius and pelvis for hematoma formation and blood loss following placement of tension-free vaginal tapes (TVT) and transobturator tapes (TOT). MATERIALS AND METHODS: 25 women were examined with ultrasonography on post-operative days one, two, three, four or five. One patient underwent TVT while the other 24 underwent TOT. Additional procedures were performed on most patients. RESULTS: No hematomas or collections of free fluid were identified post-operatively. CONCLUSIONS: TVT and TOT procedures are relatively safe procedures. We encountered no vascular complications in this small series of patients. Review of literature has yielded reports of infrequent complications. PMID- 17124914 TI - Implied contract of confidentiality between patient and physician. PMID- 17124915 TI - Chronic abdominal pain with radiological scirrhus sign. PMID- 17124916 TI - The health of Sparta, 1822. PMID- 17124917 TI - Physician participation in executions--legal and ethical considerations. PMID- 17124918 TI - Cell phones and E-mail: some thoughts. PMID- 17124919 TI - Management of papillary microcarcinoma of the thyroid. AB - Papillary carcinoma of the thyroid is usually an indolent tumor with a good survival prognosis especially when small and limited to the thyroid gland. The prevalence of this tumor in younger patients with long life expectancies, however, magnifies the problem of late recurrences. Current published guidelines recommend near total thyroidectomy for all differentiated thyroid cancers with the exception of small papillary carcinomas (< 1.0 cm) found coincidentally after surgery for benign disease, providing there are no other risk factors. Presentation of completion thyroidectomy as an option to the patient is recommended. Both the patient and physician should be aware that 15-20% of cases will have another focus in the contralateral lobe and that long-term recurrent disease may reach 20% or more in the absence of complete resection. Despite their small size, 5% or more of these tumors may be associated with invasion of the capsule or distant metastases (Stage III or IV). The task for patient and physician is to fairly balance these factors against the risks and cost of the additional procedures. PMID- 17124920 TI - Newborn screening for cystic fibrosis. PMID- 17124921 TI - Methicillin-resistant Staphylococcus aureus: a community concern. PMID- 17124922 TI - Statements of apology--a matter of ethics. PMID- 17124923 TI - Quality focus: influenza season 2006-2007. PMID- 17124924 TI - A doctor's story. PMID- 17124925 TI - Evidence-based medicine. PMID- 17124926 TI - Cholesteatoma of the external auditory canal in an immunocompromised patient. PMID- 17124927 TI - Epistaxis caused by hemangioma of the inferior turbinate. PMID- 17124928 TI - Bilateral sulcus vocalis. PMID- 17124929 TI - Follicular lymphoma. PMID- 17124930 TI - Necrotizing fasciitis of the neck. PMID- 17124931 TI - Columnar cell variant of papillary thyroid carcinoma. PMID- 17124932 TI - Severe distal esophageal stricture. PMID- 17124933 TI - Audiometry: masking. PMID- 17124934 TI - Professional service recovery audits. PMID- 17124935 TI - Role of impacted cerumen in hearing loss. AB - We performed a study to determine if cerumen in the ear canal causes significant hearing loss and to ascertain if there is any correlation between the amount of cerumen and the degree of hearing loss. Our study was conducted on 109 ears in 80 patients. The results indicated that impacted cerumen does cause a significant degree of conductive hearing loss. We found no significant correlation between the length of the cerumen plug and the severity of hearing loss. Nor did we find any significant correlation between the presence of impacted cerumen and variables such as age, sex, ethnicity, or affected side. PMID- 17124936 TI - Fibrous dysplasia of the frontal bone. AB - Fibrous dysplasia typically occurs in patients between the ages of 5 and 15 years. We report the case of a middle-aged woman with longstanding monostotic fibrous dysplasia who responded well to conservative treatment. She had presented with a very long (15 yr) history of frontal headache and pressure over the frontal sinus. On examination, a protuberance was noted in the region of the right frontal sinus. No other symptoms were evident, and findings on the ENT examination were normal. Computed tomography (CT) detected a widening of the frontal bone with osteolytic changes of the cancellous bone of the diploe. Partial multiphase skeletal scintigraphy and single-photon emission CT of the cranium showed increased bone metabolism in the area of the right frontal bone. No other hot spots were seen. In view of the absence of any complications, no tissue biopsy was performed. The patient was prescribed bisphosphonates for symptom relief. Follow-up CT 1 year later revealed no progression of the lesion. Overtreatment of fibrous dysplasia should be avoided because most patients respond to conservative management. In those cases that are refractory to medical treatment, surgery--in experienced hands--can result in a good functional and cosmetic outcome. PMID- 17124937 TI - An outpatient medical treatment protocol for peritonsillar abscess. AB - Several surgical methods are used to treat peritonsillar abscess, but no protocol for outpatient medical treatment has yet been published. Between February 2002 and February 2005, we treated 98 peritonsillar abscess patients with an outpatient medical regimen that involved hydration, antibiotics, steroids, and good pain control. All patients were Native Americans, who are known to have a particularly high incidence of peritonsillar abscess. The medical regimen was generally successful, as only 4 patients (4.1%) subsequently required post treatment needle aspiration or incision and drainage. We conclude that the medical protocol described herein provides practitioners with a viable noninvasive alternative for treating peritonsillar abscess. PMID- 17124938 TI - Malignancy in asymmetrical but otherwise normal palatine tonsils. AB - An abnormally large tonsil may be a sign of malignancy. We retrospectively analyzed the case files of 87 patients who had asymmetrically sized but otherwise normal tonsils and no risk factors for cancer to determine if asymmetry is associated with a higher incidence of malignancy. We found 2 cases (2.3%) of malignancy among these patients. One patient had high-grade non-Hodgkin's lymphoma in the larger tonsil, and the other had lymphocyte-rich Hodgkin's lymphoma. Both patients were older than 50 years, and neither had a history of recurrent tonsillitis. We believe that although the incidence of cancer in our series was small, it is significant. Therefore, we recommend routine excision of abnormally large tonsils. Moreover when making such a recommendation to a patient, it is essential that the patient have a clear understanding of the risk and benefit of having a tonsil removed solely because of asymmetry. PMID- 17124939 TI - Sarcoidosis presenting as a solitary parotid mass. AB - We describe an unusual case of sarcoidosis in which the patient presented with a discrete solitary parotid mass and no other manifestation of the disease. The diagnosis was based on the unexpected pathologic findings during examination of a superficial parotidectomy specimen. To the best of our knowledge, no such presentation has been previously reported in the English-language literature. PMID- 17124940 TI - Toxoplasmosis lymphadenitis presenting as a parotid mass: a report of 2 cases. AB - Toxoplasmosis manifesting as a parotid mass is rare; our review of the literature found only 6 previously reported cases. We report 2 new cases. Both patients presented with a small, mobile left parotid mass, and both were successfully treated with a diagnostic superficial parotidectomy. In both cases, the patient had been regularly exposed to cats and had recently eaten undercooked meat. When evaluating a parotid mass, otolaryngologists should be aware of the infectious causes of parotid swelling and lymphadenopathy and consider the possibility of toxoplasmosis when the history and pathologic findings are not suggestive of more common diseases. PMID- 17124941 TI - Polymorphous low-grade adenocarcinoma of the parotid gland. AB - Polymorphous low-grade adenocarcinoma (PLGA) of the parotid gland is rare. We describe a new case in which the patient underwent parotidectomy only to experience an extensive recurrence 2 years later. The recurrence was treated with radical surgical excision and radiation therapy, and the patient remained disease free at 5 years of follow-up. We also review the literature on primary parotid PLGA. PMID- 17124942 TI - Cutaneous angiosarcoma of the head and neck: a case presentation and review of the literature. AB - Cutaneous angiosarcoma of the head and neck is a rare vascular neoplasm. When it does occur, it is most common in elderly white men. Early diagnosis and treatment are essential for local control of this aggressive tumor but recognition can be delayed because of its rarity or because of difficulty in making a pathologic diagnosis. A combined-modality treatment approach is most often advocated. We report the case of a 77-year-old black man who presented with a 1-month history of two painless, violaceous, subcentimeter nodules of the upper lip. After a diagnosis of low-grade angiosarcoma was definitively established, the lesions were locally excised with good cosmetic and functional results. The patient subsequently was found to have probable metastatic disease, but he declined further intervention. We review the literature on cutaneous angiosarcoma, and we discuss its epidemiology, presentation, tissue diagnosis, treatment, and prognosis in an effort to increase awareness of this rare malignancy. PMID- 17124943 TI - A case of papillary carcinoma arising in ectopic thyroid tissue within a branchial cyst with neck node metastasis. AB - We describe the interesting case of a young man who presented with a lateral neck node that was diagnosed as a branchial cyst. Histopathology of the excised node revealed that a papillary carcinoma was located within thyroid tissue, which in turn was located within a branchial cyst. A total thyroidectomy with local lymph node clearance was performed. Histology identified a normal thyroid gland, but a papillary carcinoma in one of the excised lymph nodes was consistent with a metastasis. To our knowledge, this is only the second reported case of a thyroid carcinoma arising in ectopic thyroid tissue that metastasized in the neck. PMID- 17124944 TI - Blood--but not bleeding--at a tracheotomy site: a case of Munchausen's syndrome. AB - Munchausen's syndrome is a factitious illness in which patients fabricate medical problems in order to receive medical attention. Their "symptoms" are typical of actual medical conditions and their history is dramatic yet plausible, but no pathology can be found on imaging and other investigations. We report the case of a young woman whose Munchausen's syndrome manifested as purported bleeding from a tracheotomy site. In actuality, she had drawn the blood from an arm vein and spattered it on her neck. PMID- 17124945 TI - Adaptation of the application of good clinical practice depending on the features of specific research projects. AB - The conduct of clinical trials falls within a strict regulatory framework. The objective of the round table was to develop reasonable recommendations for the implementation of GCP according to the type of research and taking in account the risks and challenges related to this research. Two types of risks have been identified: those related to the characteristics of the research and those related to the impact of the study results. The group designed an evaluation table of these risks. The round table focused its investigations on 3 main themes: monitoring, the investigational medicinal product and undesirable effects. Three methods of monitoring adaptation were analysed in terms of advantages and disadvantages: the gradual approach, the central monitoring, monitoring on the basis of sampling. Examination of the investigational medicinal product focused on the medicinal product circuit. The group recommends using the following 'basic' decision-making tree, which takes three elements into account: 1) is it an investigational medicinal product?, 2) do the trial objectives and design require packaging specific to the research?, 3) is the risk of use higher than that in standard practice? Finally, adaptation of the implementation of GCP in terms of pharmacovigilance appeared very limited and could possibly be considered for the medicinal product, the subject of the research, which already holds a marketing authorisation, and for which the safety profile is well known; in this case, only simplified collection of non-serious adverse events may be envisaged, which may be implemented by designing and using a standard collection listing. The adaptation of the implementation of GCP is possible. This firstly takes into account the characteristics of the research: which objectives/which risks/which challenges. The options in terms of adaptation must be pre-defined, documented and justified; if necessary, they will also be re-assessed in the course of analysis. PMID- 17124946 TI - Risk management and monitoring methods for the future mother, embryo, fetus, and post-natal consequences. AB - Data required to asses the risk of a new drug regarding the normal course of pregnancy as well as embryo, fetal and neonate development, are often missing when a new product is launched. In such a situation, a risk management plan is to be developed by the industrial and validated by regulatory authorities. This risk management plan is to take into account the data benefits on the drug and its potential therapeutic use by women as being of childbearing age. The obtaining of post licence human data is to be built on many players, both private and public, involved in the data collection and evaluation. The setting up of such a network would allow them to join together and optimize their action by standardizing the data collected and their follow up. This should help to generate or rapidly respond to an alert, to conduct collaborative pharmacovigilance pharmacology studies. PMID- 17124947 TI - Protocolisation, use and development of anti-cancer drugs in the context of T2A (case-mix based payment system) set-up. AB - Drugs used in oncology represent more than half of the innovative and costly drugs which are not covered by a Group Homogene de Soins (DRG type classification) within the context of the case-mix based payment system (termed T2A). For these drugs, good practice reference guidelines have been drawn up by scientific societies and registration agencies. Recognised indications, relevant indications and situations where the treatment should not be prescribed are defined by the National Institute of Cancer. The reference guidelines should lead towards the good use of these drugs and allow the sick funds to control prescriptions. They should evolve with time, which means that bibliographic monitoring and independent expert opinion is necessary to update them as science provides new data. Manufacturers are involved in this process which in no case should undermine developmental efforts leading to registration. The objective of this protocolisation is to allow all patients early and legitimate access to drugs representing real therapeutic progress. These reference guidelines should be integrated into the life-cycle of a drug and should give rise to new developments allowing the good use of cancer products in situations which have been properly validated. PMID- 17124948 TI - Public-private partnership models in France and in Europe. AB - The workshop entitled "Public-Private partnerships models in Europe-- comparison between France and European countries" brought together representatives of academia and industry, of national or European health research programs, of regional or national public-private partnership (PPP) initiatives, and of biotechnology with the following objectives: sharing a common vision on the needs, expectations and challenges of public-private partnership, based on the analysis of actual and original cases, and of new initiatives on public-private partnership, drawing conclusions and identifying key success factors, identifying trails for progress and drawing recommendations. The major event in this field is a European public-private partnership initiative between pharmaceutical industry (European Federation of Pharmaceultical Industry and Associations, EFPIA) and the European Commission (DG Research--health priority) resulting in the European Technology Platform project "Innovative Medicines Initiative" (IMI). Its architecture is based on the identification of the main bottlenecks to the development of innovative treatments (predictive pharmacology and toxicology, identification and validation of biomarkers, patients' recruitment, risk evaluation, and cooperation with the regulatory authorities). Simultaneously, initiatives both at the national and regional levels also foster PPP in the therapeutic field. Regional competitivity clusters acting in the biomedical sector, and national PPP calls such as the ANR (National Research Agency) RIB (Research and Innovation in Biotechnology) call are incentives for PPP projects. These regional and national PPP levels help public and private partners to further build consortia able to compete for EU-level calls, thus acting as incubators for EU PPP projects. In spite of incentives and of the regional and national structuring of PPP, weaknesses in the French system are linked to its fragmentation--multiple transfer agencies, multiple research organisations (operator or funding agency)--making contracts more difficult. This requires a simplified organisation, with a single referent per area (health, technology...). Improvement may also result from adaptation in the carreer, recruitment and mobility, from support to scientists in the management of projects, and from consistent support (without maintaining them artificially alive) to emerging companies from concept through clinical development. Pathways have been proposed to improve the efficiency of clinical research in France and Europe, involving the public hospital sector, and this requires the connection of disease-oriented networks and integrated infrastructures in Europe. As stated in the IMI strategic research agenda on efficacy, the quality of public infrastructures in Europe will be a key factor for its competitiveness and attractiveness for both academic and industry projects. PMID- 17124949 TI - [What is the place of beta-adrenergic blockade therapy in coronary syndromes in the modern management era?]. AB - This paper shows the eventual benefits of treatment with beta-blockers in post myocardial infarction. An overview of main clinical trials that have been conducted to test this therapeutic in acute coronary syndrome. In patients presenting with acute myocardial infarction, the early use of intravenous beta blockers had shown a modest and non significant reduction in mortality. Substantial reductions in mortality and reinfarction have been demonstrated when beta-blockers have been used soon after an acute myocardial infarction and continued long-term. However, these benefits were observed in randomised clinical trials conducted in the 1970s and 1980s, prior to the widespread use of reperfusion therapies and antiplatelet agents. It appears obvious that the long term use of beta-blockers may be questionable after any acute coronary syndrome especially if complete reperfusion has been achieved and left ventricular function is preserved. PMID- 17124950 TI - Less use of NSAIDs in long-term than in recent chondroitin sulphate users in osteoarthritis: a pharmacy-based observational study in France. AB - In clinical trials, long-term use of a specific chondroitin sulphate, Chondrosult 400 (CS400) has demonstrated symptomatic efficacy in osteoarthritis comparable to that of nonsteroidal anti-inflammatory drugs (NSAIDs) with significantly fewer side-effects. CS400 could therefore reduce the use of and risks associated with NSAIDs. A cross-sectional observational study was therefore devised in 199 randomly selected pharmacies in France to verify the concomitant use of analgesic and NSAIDs medication in patients prescribed CS400. Consecutive patients filling a prescription for CS400 were prospectively recruited and classified into recent users (3 months or less of continuous use) and long-term users (more than 3 months of continuous use) of CS400. The main outcome measure was current and long term use of analgesics and NSAIDs. The 844 participating patients included 623 (73.8%) women and 221 (26.2%) men. Mean age was 65.9 years. Ninety eight (11.6%) patients did not use any analgesic or NSAIDs for osteoarthritis: 746 (88.4%) reported the use of at least one of these drugs. Compared to recent users, long term users of CS400 had a significantly lower current (44.4 versus 52.5%, p < 0.05) and long-term use of NSAIDs (11.8% versus 18.5%, p < 0.05), and of analgesics (70.3 versus 79.3%, p < 0.01). PMID- 17124951 TI - The interaction of new plant flavonoids with rat liver mitochondria: relation between the anti- and pro-oxydant effect and flavonoids concentration. AB - Flavonoids are polyphenols derivatives of plant origin that possesses a broad range of pharmacological properties, including protection of cells and tissues against the deleterious effects of reactive oxygen species. Their antioxidant activity results from scavenging of free radicals and other oxidizing intermediates, from the chelation of iron or copper ions and from inhibition of oxidases. But a number of studies have found both anti and prooxidant effects for many of these compounds. These reasons prompted us to investigate whether flavonoids compounds alone or combined flavonoids had antioxidant, free radical scavenger and antiapoptotic properties. The investigation was carried in vitro using rat hepatic mitochondria. Respiratory control ratio (RCR), oxygen consumption, adenosine tri phosphate (ATP) synthesis, scavenging action, enzymatic activities of involved complexes, superoxide anion and the release of cytochrome C were measured to assess the mechanisms of action of these drugs. Our data showed that the decrease of RCR induced by high concentrations (0.1 mM and 0.01 mM) of all flavonoids tested was due to a common inhibition of oxidative phosphorylation (State 4) and activation of state 3. At the opposite mitochondrial swelling was slightly induced only by low concentrations (10(-8) and 10(-9) M) of the flavonoids. They had no effects on the mitochondrial complexes (I to V) activity. Furthermore the mitochondrial membrane potential was not affected by any flavonoids. The effect of flavonoids on superoxide anion generation was variable. All the flavonoids studied acted between 10(-4) M and 10(-6) M with no effects at lower concentrations. These effects were similar on lipid peroxidation (malondialdehyde [MDA] levels). We remarked a concentration dependent in the effect of flavonoids since they acted as antioxidant and also as uncoupler at high concentrations, which is a risk for the cells. We conclude that flavonoids extracted from algerian plants have some protecting effects against oxidative stress by protecting the mitochondria. PMID- 17124952 TI - [Depersonnalization with beta 1b interferon]. PMID- 17124953 TI - [Severe and unusually prolonged acute poisoning with primidone suggesting the presence of bezoars]. PMID- 17124954 TI - [Avascular osteonecrosis of the jaws induced by diphosphonates]. PMID- 17124955 TI - [Biliary lithiasis and cholecystitis with voriconazole: about 3 cases]. PMID- 17124956 TI - Lack of cultural competency: a business and clinical risk. PMID- 17124957 TI - Structuring communication for team-based error management. PMID- 17124958 TI - Meconium in the amniotic fluid: a risk manager's perspective. PMID- 17124959 TI - System technology can improve management of product alerts. PMID- 17124960 TI - General liability. Hospital may have duty to control rowdy children. Rodriguez v. 1201 Realty LLC, 781 N.Y.S. 2d 328 (Sup. Ct. NY 2004). PMID- 17124961 TI - Vicarious liability. Hospital not liable for actions of independent practitioners in Michigan. Inglis v. Providence Hospital and Medical Centers, Inc.,__ Mich. App.__, __ N.W. 2d __, No. 247066 (Mich. Ct. App. August 26, 2004). PMID- 17124962 TI - Comparative negligence. Negligence of the plaintiff does not offset damages. Mercer v. Vanderbilt University, Inc., 134 S. W. 3d 121 (Tenn. 2004). PMID- 17124963 TI - Provider/patient relationships. Cases demonstrate the nature of the provider/patient relationship. Wilson v. Athens-Limestone Hospital, __ So. 2d __, No. 1030013 (Ala. June 18, 2004); Roberts v. Sankey, 813 N.E. 2d 1195 (Ct. App. Ind. 2004). PMID- 17124964 TI - Professional/general liability. Provision of care may have mixed liabilities. Bryant v. Oakpointe Villa Nursing Centre, 684 N.W. 2d 864 (Mich. 2004). PMID- 17124965 TI - Multiple sclerosis--what occupational health nurses need to know. PMID- 17124966 TI - Use of a violence risk assessment tool in an acute care hospital: effectiveness in identifying violent patients. AB - This study examined the use and effectiveness of the Alert assessment form. The form is part of the Alert system, used by one large acute care hospital to identify patients with a propensity for violence. All reported incidents of patient violence from August 1, 2003, through December 31, 2004, were included in patient charts. One hundred seventeen violent patient charts were reviewed and compared with 161 non-violent patient charts, randomly chosen from the same time period. Overall use of the Alert assessment form for violent and non-violent patients was 75.7% and 35.4%, respectively. The assessment form was found to have moderate sensitivity (71%) and high specificity (94%). It is reasonably effective in identifying potentially violent or aggressive patients when it is used according to protocol. Efforts to improve the tool are warranted, as is evaluation of its benefit in settings with low prevalence of violence. Also, greater effort must be taken to prevent violence once an aggressive patient has been identified. PMID- 17124967 TI - Environmental justice: implications for occupational health nurses. AB - Through the use of innovative tools, such as clinical mnemonics, exercises in risk and asset mapping, and strategic program development, occupational health nurses can incorporate dimensions of environmental justice (EJ) into the workplace. Occupational health nurses who also take on educational roles can use case studies and network with labor and EJ groups to provide clinical experiences for occupational and environmental health nursing students, thereby integrating EJ into occupational and environmental health nursing practice. Occupational health nurses are well positioned to serve as technical experts within community based participatory research projects. Occupational health nurses must share their knowledge and experience as members of coalitions that represent workers in their fight for worker health and safety. PMID- 17124968 TI - Developing policies and procedures. AB - The development of policies and procedures is an integral part of the occupational health nurse's role. Policies and procedures serve as the foundation for the occupational health service and are based on its vision, mission, culture, and values. The design and layout selected for the policies and procedures should be simple, consistent, and easy to use. The same format should be used for all existing and new policies and procedures. Policies and procedures should be reviewed periodically based on a specified time frame (i.e., annually). However, some policies may require a more frequent review if they involve rapidly changing external standards, ethical issues, or emerging exposures. PMID- 17124969 TI - Posttraumatic stress disorder after the hurricanes. PMID- 17124970 TI - To clone alone: the United Nations' Human Cloning Declaration. AB - The United Nations labored for almost four years to create a treaty governing human cloning. In 2005 that effort was abandoned, and instead the United Nations' General Assembly adopted a "Declaration on Human Cloning". PMID- 17124971 TI - [Protection of genetic data in Spain. Analysis based on the general principles of personal data protection]. AB - The genetic data is Spain is not regulated specifically, rather, we must look at the regulation on the protection of data of a personal nature. This is turn, establishes a series of general principles to apply to any type of data. Analysing this with other regulations that are dispersed both in the national and international regulations, we can deduce the rights and obligations in this field. This highlights the fact that one can't dispose of the genetic data in the same manner as the personal data. PMID- 17124972 TI - Human genome and protection of human rights in Croatia. AB - In the context of the human rights, human genome should be analyzed on two different levels. First one is a global macro level mostly determinated by global international documents and the concept of the human genome as a common biological heritage of all human beings. The second level is a private micro level of every human individual whose human genome is expressed by specific DNA "bar code". Information of that personal identity card should be strictly protected through the legal instruments for the protection of privacy rights. The aim of this paper is to analyze if and how human rights are protected within the legal framework and the practice of DNA testing in Croatia. PMID- 17124973 TI - [The legal question of the obtention of human stem cells for biomedical research. Legislation policy considerations]. AB - The future Law on Biomedical Research, whose draft bill has been approved by the Council of Ministers and that will soon begin its parliamentary process of approval, will regulate, among other matters, the research with embryos. Likewise, it will make a pronouncement on the so-called therapeutic cloning. This report makes a detailed analysis of different matters that must be borne in mind by the legislator in order to face the process of evaluation and approval of said Law in relation with the aforementioned matters. It makes a special analysis of the legal texts of an international nature to which Spain is unavoidably subjected to, in such a way that the legislative text that will finally be approved is not contrary to the dispositions that are within such. PMID- 17124974 TI - [The Evans case and the right not to be forced to reproduce]. AB - The article analyses the ruling of the European Court of Human Rights, Evans v. United Kingdom, of March 7, 2006 (application no. 6339/05) from the perspective of the nature and the effects of a consent together with the conservation and disposal of cryopreserved embryos. Several prevailing normative models are studied in order to look at the disposal of embryos and to test its legal consistency. As a conclusion and basing it on the Evans case, there is a proposition to distinguish between the acts of disposal of spare embryos from IVF programs and the deposit of embryos as a way to prevent lack of fertility. PMID- 17124975 TI - [The 14/2006 law on human assisted reproduction techniques: scientific and ethical considerations]. AB - The new Spanish Law on Artificial Human Reproduction Techniques is analyzed from the scientific, ethical and legal points of view, paying special attention to the preimplantational diagnosis and the experimental utilization of gametes and preembryos. Other items are also analyzed. PMID- 17124976 TI - [The status of human cloning in the international setting]. AB - The General Assembly of the United Nations submitted a Declaration on Human Cloning in March 2005. The text of such Declaration was the result of a difficult and long process, taking more than three years. Being a Declaration instead of a Resolution, it has not legal capability in inforcing United Nations members to act according to its recommendations. This article begins with an explanation of several terms referred to cloning. Different countries' legislation on cloning is analyzed. Positions of the same countries at the Convention of the United Nations are as well analyzed. Comparing both countries' views shows that national legislation on cloning is independent and orientated by some countries' particular interests and biological and ethical views on these issues. Future developments on human cloning and its applications will be shared among all countries, both the ones currently allowing and supporting "therapeutic" cloning and the ones now banning it. In such case, it would be important to reach agreements on these issues at an international level. The article discusses possible legislative developments and offers some proposals to reach such agreements. PMID- 17124977 TI - Science and the courts: a fruitful experiment of interaction. AB - The authors report on the European Seminar on Hot Genetic Issues and the Courts, held in Pavia (I), September 2005. They examine the new possible concerns regarding bio-sciences practical applications and the stronger need for a closer interaction between jurists, judges and scientists. They highlight the importance of the ENLSC's networking experience in judicial education and the originality of its approach. PMID- 17124978 TI - [The 14/2006 law, of May 26, on human assisted reproduction techniques]. PMID- 17124980 TI - References to books, articles in periodicals and to other documents. PMID- 17124979 TI - An international consortium on stem cells, ethics and law. PMID- 17124981 TI - Knowledge and use of preventive measures against malaria in endemic and non endemic villages in northern Thailand. AB - The objective of this study was to determine the frequency and determinants of knowledge of malaria in four sites in northern Thailand, and to compare the use of prevention measures between people with and without a knowledge of malaria. An epidemiological survey was conducted in January 2002 among 857 persons living in four sites in northern Thailand. Of the 857 persons, 53% had a knowledge of malaria, ranging from 38% and 51% in non-endemic to 76% and 77% in endemic sites. Headache (89%), shivering (83%) and fever (79%) were the most frequently mentioned symptoms. Younger persons had more knowledge of malaria than older persons; adjusted Odds Ratio (aOR): 3.91 [95% confidence interval (CI): 2.32 6.56] for the 15-29 age group compared to persons 60 years and older. In comparison to men, knowledge of malaria was significantly lower among women (aOR: 2.00, 95%-CI: 1.47-2.70). Persons with knowledge reported a significantly higher use of prevention measures than persons without knowledge of malaria. PMID- 17124982 TI - Riboflavin-deficient and Trichinella spiralis-induced stresses on plasma corticosterone associated with spermatogenesis in male Wistar rats. AB - The objective of this study was to investigate the effects of riboflavin deficient and Trichinella spiralis-induced stresses on corticosterone associated with spermatogenesis in male Wistar rats. Rats were allocated into 4 groups: Group 1: control; group 2: riboflavin-deficient diet; group 3: T. spiralis infection; group 4: riboflavin deficient diet with T. spiralis infection. This experiment lasted for 12 weeks. Plasma corticosterone was significantly enhanced when exposed to acute riboflavin deficiency and/or T. spiralis infection stress. When the rats were chronically subjected to such stresses, T. spiralis per se had prolonged effects, in a marked increase in corticosterone. T. spiralis per se tended to impact on such sperm characteristics as sperm motility, sperm count and daily sperm production, even defected seminiferous tubules. It was proposed that the Trichinella spiralis-induced stress probably had adverse effects on the level of adrenocortical-testicular axis whenever their habitats on muscle fibers were evident. However, riboflavin-deficient-induced stress had little implication in the adrenocortical-testicular axis. PMID- 17124983 TI - Novel real-time PCr for detection of Schistosoma japonicum in stool. AB - Chemotherapy has been used on a large scale in countries where the blood fluke Schistosoma japonicum is endemic. This has led to a lower intensity of infections and consequently lower diagnostic values of commonly used diagnostic tests like serology and Kato-Katz stool smear. We designed a novel real-time PCR method for detection of S. japonicum in stool samples. Further, we evaluated different versions of an inexpensive, non-commercial extraction method, ROSE, as well as the commercial QIAamp DNA Stool Mini Kit. PCR primer sequences were designed targeting the mitochondrial NADH dehydrogenase I gene. Bovine serum albumin was added to the DNA extracts and SYBR Green was used for detection. The PCR method was evaluated with non-infected stool samples spiked with S. japonicum eggs. It demonstrated high sensitivity, even in samples containing a single egg. The two extraction methods were equally effective. The PCR was specific for S. japonicum when tested against other Schistosoma species, Trichuris trichiura, hookworm and Taenia sp. We conclude that this novel real-time PCR, in combination with either ROSE or QIAamp DNA Stool Mini Kit extraction, is a sensitive and specific tool for diagnosing S. japonicum in human stool samples. PMID- 17124984 TI - Formulation of tablets from the crude extract of Rhinacanthus nasutus (Thai local plant) against Aedes aegypti and Culex quinquefasciatus larvae: a preliminary study. AB - Dried root powder of Rhinacanthus nasutus, Thong Phan Chang (Thai name) were extracted with methanol (MeOH) in a Soxhlet apparatus and made into 2 formulations of tablet containing the extract at 5% and 10% concentration. Due to the viscous and poor flow properties of the crude MeOH extract obtained, a wet granulation method was conducted in developing the tablets. Lactose was used as a filler. Polyvinyl pyrrolidone (PVP) K30 (15% w/w solution in alcohol) was used as the binding agent, while stearic acid (2% w/w) was used as a lubricant. Both formulas of prepared tablets had a smooth shiny surface with a round shape. Other physical properties of the tablets, such as weight variation, friability and disintegration time, met the requirements of the USP XX standard. The mosquito larvicidal activity of prepared tablets containing 5% and 10% R. nasutus extract against Aedes aegypti were not significantly different from each other (p > 0.05), with 48-hour LC50 values of 13.6 and 14.2 mg/I for the 5% and 10% tablets, respectively, while their activities against Culex quinquefasciatus were similar (p > 0.05) with LC50 values of 18.7 and 17.3, respectively. The larvicidal activity levels against Ae. aegypti and Cx. quinquefasciatus were also not significantly different from each other (p > 0.05). No larval mortality was observed in the two control groups: lactose solution and dechlorinated water. Toxicity to female and male fish (Poecilia reticulata) was tested with the prepared tablets. The toxicity of tablets containing 5% and 10% extracts were not significantly different from each other for the P. reticulata females with 48 hour LC50 values of 105.2 and 110.8 mg/I, respectively, and for P. reticulata males with LC50 values of 99.1 and 103.4 mg/I, respectively. Female and male P. reticulata were sensitive to the same dose of the extract. No fish died in the two control groups, with lactose solution and dechlorinated water. Acute-toxicity bioassay with fish showed that with an exposure of 48 hours the LC50 values of the tablets containing 5% and 10% were 5- to 10-fold higher than the LC50 of R. nasutus against mosquito larvae. These prepared tablets could possibly used to control mosquito vectors and be introduced into the mosquito control program. PMID- 17124985 TI - Bionomics studies of Mansonia mosquitoes inhabiting the peat swamp forest. AB - The present study was conducted in the years 2000-2002 to determine the bionomics of Mansonia mosquitoes, vectors of nocturnally subperiodic Brugia malayi, inhabiting the peat swamp forest, "Phru Toh Daeng", Narathiwat Province, Thailand. Fifty-four species of mosquitoes belonging to 12 genera were added, for the first time, to the list of animal fauna in the peat swamp forest. Mansonia mosquitoes were the most abundant (60-70%) by all collection methods and occurred throughout the year with a high biting density (10.5-57.8 bites per person-hour). Ma. bonneae was most prevalent (47.5%) and fed on a variety of animal hosts, including domestic cats, cows, monkeys, and man with a maximum biting density of 24.3 bites per person-hour in October. The infective bites were found for the first time in Ma. annulata collected at Ban Toh Daeng (13 00-14 00 hours) and also Ma. bonneae at forest shade (16 00-17 00 hours) and in a village (20 00-21 00 hours) with rates of 0.6, 1.1 and 1.0%, respectively. The biting activities of these two species occurred in both the day and night time, with two lower peaks at 10 00 hours (18.5 bites per person-hour) and 13 00-15 00 (8.5-10.0 bites per person-hour) hours, but the highest peak was 19 00-21 00 hours (31.5-33.0 bites per person-hour) The biting activity patterns corresponded with the periodicity found in man and domestic cats and may play an important role in either transmission or maintenance of the filarial parasites in the peat swamp forest. The relative role of Ma. bonneae and Ma. uniformis in different environmental settings (primary swamp forest and open swamp) on the transmission of nocturnally subperiodic B. malayi merits further study. PMID- 17124986 TI - Surveys for natural host plants of Mansonia mosquitoes inhabiting Toh Daeng peat swamp forest, Narathiwat Province, Thailand. AB - Surveys were carried out monthly from April-October 2002 to examine 68 sampling sites around "Toh Daeng" peat swamp forest in Narathiwat Province, Thailand, of which 38 were known Mansonia-positive habitats and 30 were Mansonia-negative sites. The present larval surveys were qualitative owing to features of the host plants (location, distribution, and abundance), difficulties in locating and selecting the host plants in the swamp forest, and time constraints. Twenty attempts were made for each species for larvae. The presence of Mansonia larvae on each plant species was confirmed 6 times for each plant and location. Larvae of Ma. bonneae and Ma. uniformis were obtained from 18 plant species (10 families): Metroxylon sagu, Melaleuca cajuputi, Pandanus militaris, Pandanus immerses, Hanguana malayana, Typha angustifolia, Hymenachne acutigluma, Scirpodendron ghaeri, Scleria sumatrensis, Rhynchospora corymnosa, Sacclolepis indica, Cyperus babakan, Cyperus corymbosus, Lepironia articulata, Leersia hexandra, Eichhornia crassipes, Pistia stratriotes and ferns. The emergent grasses, S. ghaeri, S. sumatrensis, H. acutigluma, R. corymnosa, S. indica, C. babakan, C. corymbosus, and L. articulata, were the preferred host plants. Samples from larger trees, M. sagu and M. cajuputi, yielded low numbers of 1-7 larvae per scraping. Ma. uniformis was recovered from most of the host plants, while Ma. bonneae preferred submerged plants and was not found on the floating aquatic plants, E. crassipes and P. stratriotes. The description of modified dipper and dipping techniques are given and discussed. PMID- 17124987 TI - Relationship between body size and severity of dengue hemorrhagic fever among children aged 0-14 years. AB - A hospital based case-control study was conducted from October 2002 to November 2003 among children aged 0-14 years at Queen Sirikit National Institute of Child Health (Children's Hospital), Bangkok, Thailand. This study focused on body size and severity of dengue hemorrhagic fever (DHF) in children. One hundred five patients diagnosed as having DHF grade III or IV were the cases and 105 diagnosed as having DHF grade I or II were controls. They were matched at a ratio of 1:1 by their gender and age (within 5 years). Normal growth charts were used to differentiate child body size into normal, thin and obese. Data were collected using face to face interviews with caregivers, questionnaires, laboratory and physical examination reports as research tools. Multiple logistic regression analysis revealed that only two variables were related to severity of DHF: obesity (OR = 3.00, 95 % CI = 1.20-7.48) and dengue virus type II (OR = 4.94, 95 % CI = 2.57-9.47), respectively. Other variables were childhood factors: duration of breast-feeding, education, and parity; caregivers factors: age, gender, marital status, education, occupation, family income, knowledge of DHF, antipyretic type, treatment before hospitalization, and duration of fever; environmental factors: history of DHF patients in house, house pattern, time from house to hospital, and residence; and etiological factors: type of infection and history of DHF among children. These factors showed no significant association (p > 0.05). This result can be utilized in a preventive and control program, particularly in more aggressive management of overweight children. Health personnel should continue to provide health education, particularly, signs and symptoms of shock, to the community and private sectors. Government and Non Government Protective Projects in primary schools (5-9 years children) should be continued in the high risk groups. PMID- 17124988 TI - High prevalence of HGV coinfection with HBV or HCV among northeastern Thai blood donors. AB - Hepatitis G viral (HGV) infection among northeastern Thai blood donors was determined by the nested RT-PCR technique. HGV RNA was amplified by the degenerated helicase primers for a product of the expected size of 83 base pairs were used in this study. Serum samples from 322 of three different categories of northeastern Thai blood donors were included in this study. There were 104 HBsAg and Anti-HCV seronegative blood donors (control group), 100 samples of HBs Ag seropositive blood donors (HBV infected group) and 118 serum samples from anti HCV seropositive blood donors (HCV infected group). The results demonstrated that HGV RNA was not detected in the control group but was found in 10 individuals (10%) in the HBV infected group and 13 (11%) in the anti-HCV positive blood donors. The prevalences of HGV in both seropositive groups were significantly different from the control group (p = 0.001). HGV co-infection is highly prevalent among northeastern Thai blood donors who are infected with HBV or HCV. The results also reveal that blood donors seronegative for HCV and HBV are a low risk group for HGV infection. PMID- 17124989 TI - The prevalence of human cytomegalovirus seropositivity among blood donors at the Unit of Blood Transfusion Medicine, Hospital Universiti Sains Malaysia. AB - Human cytomegalovirus (HCMV) is a species-specific DNA virus of the Herpetoviridae family. After a primary infection, HCMV persists in a latent form most probably in bone marrow progenitor cells or in peripheral blood monocytes. The virus can reactivate to result in shedding of the virus leading to virus dissemination and new infections. Immunocompromized patients are the ones most vulnerable to serious diseases occasionally acquired in blood transfusions. In a human population, HCMV seropositivity increases steadily with age to become approximately 100% in adults. This study was performed to detect seropositivity among regular blood donors in The Hospital of the Universiti Sains Malaysia, in the state of Kelantan. Using an enzyme immunoassay, it was found that 97.6% of blood donors were HCMV-positive. HCMV is highly prevalent and may be endemic in Kelantan. Hence, long-term strategies are required for the reduction of disease dissemination, and to prevent the exposure of immunocompromized patients to the virus. PMID- 17124990 TI - Development of a duplex-polymerase chain reaction for rapid detection of pathogenic Leptospira. AB - A duplex-polymerase chain reaction (PCR) for the rapid detection of pathogenic leptospires was developed by using two sets of newly designed primers which amplified in the same reaction two different DNA fragments simultaneously: 279-bp of LipL32 and 430-bp of 16S rRNA. For DNA extraction from bacterial cultures, the silica-based spin column method was found to be more suitable and was selected for the extraction of DNAs from all 92 bacterial strains including 56 strains of pathogenic Leptospira, 15 strains of non-pathogenic Leptospira and 21 other strains of bacteria. The PCR products were analyzed by agarose gel electrophoresis with confirmation by Southern and dot hybridization using synthetic DNA probe prepared from LipL32 gene of a pathogenic reference strain, L. interrogans serovar pyrogenes. The duplex-PCR allowed detection of two products of 279 bp and 430 bp in all pathogenic Leptospira. Non-pathogenic Leptospira generated a single product of 430 bp. Other bacterial strains failed to reveal any amplification products. As little as 1 pg of pure DNA corresponding to 100 cells could be detected by agarose gel-electrophoresis, and 1-10 fg of pure DNA by hybridization. PMID- 17124991 TI - Western immunoblot analysis using a ten leptospira serovars combined antigen for serodiagnosis of leptospirosis. AB - Leptospirosis is a major zoonotic disease throughout the world. There are unavailable accuracy diagnostic methods for the acute phase of infection. To demonstrate the advantage of Western immunoblot, a mixed leptospira serovars antigen for the serodiagnosis of leptospirosis was employed. SDS-PAGE and Western immunoblot was performed using a 10 mixed leptospira serovars antigen and stained with 16 reference rabbit anti-leptospirosis antibodies. The result showed different immunoreactive band patterns for each reference serum. The bands with molecular weights of 15-20, 23-24, 41 and 45 kDa were commonly found (88% to 100% of the 16 reference sera). Using combined leptospira antigens in a Western immunoblot technique is an alternative and practical strategy for a more sensitive leptospirosis serodiagnosis. PMID- 17124992 TI - Sensitivity and specificity of an in-house rapid urease test for detecting Helicobacter pylori infection on gastric biopsy. AB - We developed an in-house rapid urease test (iRUT) and evaluated the efficacy and the agreement of the iRUT and the cRUT compared with culture and histology for the detection of H. pylori infection. Five iRUT media were tested with H. pylori isolates and other bacteria. The most suitable iRUT medium was further evaluated for detection of H. pylori infection. Gastric biopsies from 120 patients were diagnosed by culture, iRUT, cRUT and histology. The results of the iRUT and cRUT were read at 30 minutes, 1 hour and up to 24 hours. A true positive result was either the culture or both the RUT (cRUT or iRUT) and the histological examination being positive. The sensitivity and specificity of the iRUT result at 30 minutes, 1 hour and up to 24 hours were 77.1% and 100%, 77.6% and 100%, and 94.1% and 94.2%, respectively. Values for the same parameters of cRUT were 87.5% and 100%, 89.8% and 100%, and 100% and 94.2%, respectively. The agreement between the iRUT and cRUT was very good (kappa values > or = 0.82). Our results indicate that the iRUT is a-sensitive, specific and cost effective test. It can be appropriately applied for detecting H. pylori infection in gastric biopsy specimens. PMID- 17124993 TI - Risk factors and clinical outcomes of penicillin resistant S. pneumoniae community-acquired pneumonia in Khon Kaen, Thailand. AB - To determine the prevalence, risk factors and clinical outcomes of penicillin resistant S. pneumoniae (PRSP) in community-acquired pneumonia (CAP), a cross sectional study was conducted between January 1995 and December 2004 at Srinagarind Hospital, Khon Kaen, Thailand. Patients hospitalized with CAP and culture proved to be S. pneumoniae were included. PRSP was found in 22 of 64 (34.4%) patients. The MIC levels of penicillin non-susceptible strains ranged between 0.25 and 0.75 microg/ml. Resistance to other antibiotics ranked: cotrimoxazole (51.6%), tetracycline (26.6%), erythromycin (20.6%), lincomycin (18.7%), chloramphenicol (12.5%) and ampicillin (1.6%). None of the isolates was resistant to cephalothin. The significant risk factors for PRSP infection were previous antibiotic use within 3 months (Adjusted OR 40.83, 95% CI 3.71 to 449.41) and alcoholism (Adjusted OR 8.82, 95% 1.25 to 62.46). Bacteremia and empyema thoracis were found more commonly in PRSP than PSSP infection, but not statistically significant. Pneumonia-related mortality was nearly the same, PRSP 9.1% vs PSSP 9.5% (p = 0.96). The reason why the clinical outcomes of these two groups were not different may be the patients were infected with mildly resistant organisms. Thus, pneumonia caused by intermediate-level penicillin resistant S. pneumoniae appears to be adequately treated with beta-lactams or aminopenicillin antibiotics. The MIC levels of penicillin resistance should be monitored further. The need for antibiotics active against drug-resistant S. pneumoniae was required if high-level penicillin resistance was detected. PMID- 17124994 TI - Discriminatory powers of molecular typing techniques for methicillin-resistant Staphylococcus aureus in a university hospital, Thailand. AB - Discriminatory powers of various molecular techniques were evaluated for typing of methicillin-resistant Staphylococcus aureus (MRSA) isolated in Siriraj Hospital, Bangkok, Thailand. Thirty MRSA isolates were randomly selected in this study. They were characterized by pulsed-field gel electrophoresis, Clal-mecA and Clal-Tn554 polymorphisms, ribotyping, and PCR-based methods including SCCmec typing, spa and coa gene polymorphism, and repeat units in hypervariable region downstream of mecA. Individual molecular typing technique distinguished those MRSA isolates into 2 to 5 types. Eleven genetic backgrounds of MRSA isolates were elucidated by combination of typing methods with trimethoprim/sulfamethoxazole (TMP/SXT) susceptibility. Combination of all typing methods including TMP/SXT susceptibility yielded a discriminatory index of 0.94. Combination of PCR-based methods and TMP/SXT susceptibility, with the discriminatory index of 0.89, is a practical typing approach suitable for rapid epidemiological investigation of MRSA isolates in a hospital setting. PMID- 17124995 TI - Case report vancomycin -resistant Enterococcus faecium VanA phenotype: first documented isolation in India. AB - In recent years, vancomycin-resistant enterococci, especially Enterococcus faecium has emerged as an important nososcomial pathogen and represents a serious threat to patients with impaired host defences. We report infection with vancomycin-resistant Enterococcus faecium in a 3-year-old child with patent ductus arteriosus. The organism, isolated from a central venous catheter tip, exhibited a high-level resistance to vancomycin (minimum inhibitory concentration > or = 256 microg/ ml) and was also resistant to teicoplanin. The child probably died due to sepsis from this highly resistant organism. To the best of our knowledge, this is the first reported isolation of VanA phenotype Enterococcus faecium in India. PMID- 17124996 TI - Medical characteristics and tuberculosis treatment outcomes in an urban tertiary hospital, Thailand. AB - The purpose of this study was to describe medical characteristics and assess treatment outcomes at a respiratory clinic, tertiary hospital, Bangkok, Thailand. One hundred seventy-seven patients diagnosed as having any type of TB and having taken anti-TB drugs for one month were included in the study. The patients were interviewed the end of the first month and their medical files were reviewed at the end of treatment to obtained the treatment outcome. Descriptive statistics and chi-square test were used in the data analyses. Of the total of 177 patients, 33.3% were new smear-positive, 19.2% were new smear-negative, 18.1% were extra pulmonary TB, and 29.4% were other. The prevalence of TB/HIV co-infection was 33% (18 of 54). Resistance to at least one anti-TB drug was 31.6% (6 of 19). Due to high treatment default rates, treatment completion rates were low, 64.4% in new smear-positives, 61.8% in new smear-negatives, 71.9% in extra-pulmonary TB, and 46.2% in other. Treatment default rates significantly differed among the four types of TB (chi2 = 8.3, p = .04). The findings indicate a high proportion of extra-pulmonary TB, high prevalence of TB/HIV coinfection, and low treatment completion in urban TB patients at the tertiary hospital. Interventions are needed to integrate the strengths of the tertiary hospital regarding the availability of chest specialists and advance diagnostic tools into the other levels of health service to improve treatment outcomes in urban populations. PMID- 17124997 TI - Missed appointments at a tuberculosis clinic increased the risk of clinical treatment failure. AB - We investigated the charts of 381 new smear-positive tuberculosis patients at Khon Kaen Medical School during 1997-2001 using World Health Organization definitions to evaluate associations among treatment success or failure (defaulted, failed, died, or not evaluated) and tuberculosis clinic contact, demographics and clinical characteristics of the patients. Multinomial logistic regression was used for three-category outcome analysis: treatment success, transferred-out and clinical treatment failure. The treatment success and clinical treatment failure rates were 34.1% and 34.4%, respectively. About 46.5% and 85.8% of patients missed appointments at the tuberculosis clinic in the treatment success and treatment failure groups, respectively. The results show that patients who were absent from the tuberculosis clinic were 5.95 times more likely to have clinical treatment failure than treatment success, having adjusted for the effect of transfering-out and the effect of the treatment regimen and the sputum conversion status (adjusted odds ratio = 5.95; 95% CI: 2.99 to 11.84). The review showed that absence from the tuberculosis clinic was an independent risk factor for clinical treatment failure. We recommended that all new smear-positive tuberculosis patients should be followed closely at a tuberculosis clinic. PMID- 17124998 TI - A hospital-based study of bloodstream infections in febrile patients in Dhulikhel Hospital Kathmandu University Teaching Hospital, Nepal. AB - The etiology of bloodstream infections in febrile patients remain poorly characterized in Nepal. A retrospective study of febrile patients presenting to Dhulikhel Hospital Kathmandu University Teaching Hospital from July 2002 to June 2004 was performed to evaluate the etiology of bloodstream infections and the drug sensitivity patterns of cultured organisms. The medical and laboratory records of all febrile patients with an axillary temperature > or = 38 degrees C who had a blood culture taken (n = 1,774) were retrieved and analyzed. Of these, 122 (6.9%) patients had positive blood cultures, of which 40.1% were age 11 to 20 years. The male to female ratio was 1.7:1. Antibiotics had been taken prior to hospital presentation by 39 (32%) patients. Salmonella enterica serovar Typhi and serovar Paratyphi A were isolated in 50 (41.0%) and 13 (10.7%) cases, respectively. All S. Typhi and S. Paratyphi isolates were susceptible to ceftriaxone, while susceptibility to ciprofloxacin and chloramphenicol was recorded in 94.8% and 94.5% of cases, respectively. Cephalexin and amoxicillin had the lowest rates of susceptibility (64.2% and 54.1%, respectively). Salmonella spp were usually sensitive to chloramphenicol. These findings provide clinicians in this region of Nepal with a better understanding of the spectrum of pathogens causing bloodstream infections and will help guide empiric antibiotic choice. PMID- 17124999 TI - HLa-class II (DRB & DQB1) in Thai sudden unexplained death syndrome (Thai SUDS) families (Lai-Tai families). AB - Thai Sudden Unexplained Death Syndrome (Thai SUDS), or Lai-Tai, is a major health problem among rural residents of northeastern Thailand. The cause has been identified as a genetic disease. SUDS, a disorder found in Southeast Asia, is characterized by an abnormal electrocardiogram with ST-segment elevation in leads V1-V3, identical to that seen in Brugada Syndrome (Brugada Sign, BS) and sudden death due to ventricular fibrillation and cardiac arrest (represents an arrhythmogenic marker that identifies high-risk for SUDS). SUDS victims have a sleeping disorder (narcolepsy). The HLA-DR locus is tightly associated with narcoleptic Japanese patients and HLA-DR2, DQ haplotypes were also found in Oriental narcoleptic patients. These circumstances prompted us to study the association between the disease and HLA Class II by HLA DNA typing using a PCR SSO method, with five Thai SUDS families (18 BS-positive subjects as the cases, and 27 BS-negatives as the controls). We found that the HLA-DRB1 *12021 allele was significantly increased in BS-positive subjects (p = 0.02; OR = 4.5), the same as the HLA-DRB1*12021-DQB1 *0301/09 haplotype (p = 0.01; OR = 7.95). Our data suggests that the HLA-DRB1* 12021 allele associated with BS and the HLA DRB1*12021-DQB1 *0301/09 is a haplotype susceptible to arrhythmogenic markers that can identify a high risk for SUDS. PMID- 17125000 TI - Risk of a couple having a child with severe thalassemia syndrome, prevalence in lower northern Thailand. AB - Thalassemia screening in pregnant women and their spouses was performed at Buddhachinaraj Provincial Hospital and 8 community hospitals in Phitsanulok; lower northern Thailand. The prevalence of thalassemic carrier state was determined of 1,198 couples. Of these, 4.8% had heterozygous alpha thalassemia-1, 1.6% had heterozygous beta thalassemia, 12.4% had heterozygous hemoglobin (Hb) E, 2.7% had homozygous Hb E and 0.25% of others had abnormal Hb. Eighteen at risk couples (1.5%) were identified. Fifteen couples were at risk for compound heterozygous Hb E / beta thalassemia and the remaining 3 were at risk for homozygous alpha thalassemia-1. Prenatal diagnosis (cordocentesis) was performed in 4 couples at risk, but no fetuses with severe thalassemic disease were detected. PMID- 17125001 TI - A study of serum ferritin levels among male blood donors in Hospital Universiti sains Malaysia. AB - Iron deficiency is the commonest cause of anemia worldwide and healthy blood donors are estimated to lose about 236 mg of iron with each donation. The objective of this study was to determine the serum ferritin levels among first time and regular male blood donors, and also to correlate the serum ferritin levels with the number of donations and hemoglobin levels. Hemoglobin levels and serum ferritin were measured in three groups of donors divided into first time donors; (n = 92), donors with 2-4 donations (n = 41), and regular donors (n = 78). The mean hemoglobins in the first time donors, second group and regular blood donor group were 14.95 +/- 1.08, 15.12 +/- 1.44 and 15.56 +/- 1.48, respectively. The serum ferritin level were found to be significantly lower among the regular donors (62.0 +/- 39.78 ng/ml) compared to first time donors (90.7 +/- 66.63) and second group donors (114.12 +/- 66.97). The serum ferritin levels gradually decrease according to the number of donations and there was a significant correlation between frequency of donations and the serum ferritin level (r2 = 0.082). Significant correlation between the number of donations and hemoglobin level r2 = 0.061) was noted. However, there was no significant correlation between hemoglobin and serum ferritin levels (r2 = 0.015). Eleven percent of regular donors had depleted iron stores. This was not noted in donors who donated less than 5 times within 2 years. PMID- 17125002 TI - C-reactive protein, interleukin-6, and tumor necrosis factor-alpha levels in overweight and healthy adults. AB - This study aimed to 1) compare levels of high sensitivity c-reactive protein (hs CRP), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) between overweight Thais and apparently healthy controls, and 2) investigate the association between serum hs-CRP, IL-6, and TNF-alpha levels and other biochemical parameters. A total of 180 health-conscious adults aged 25-60 years, who resided in Bangkok, participated in this study. No significant difference was found in age and sex between the overweight subjects and controls. Serum levels of hs-CRP, IL-6, TNF-alpha, glucose, lipid profile, body mass index (BMI), waist circumference (WC), hip circumference (HC) and waist hip ratio (WHR) were determined in these volunteers. The mean levels of white blood cells (WBC), uric acid, total cholesterol (TC), triglyceride (TG), and hs-CRP were significantly higher in the overweight subjects than those in the controls, whereas high density lipoprotein-cholesterol (HDL-C) values were significantly higher in the controls than the overweight subjects (p < 0.05). Hs-CRP levels were significantly positively correlated with levels of TG, BMI, WC, HC and WHR. HDL-C levels were significantly negative correlated with hs-CRP levels. In conclusion, the prevalence of elevated serum hs-CRP levels was higher in overweight subjects than controls. However, more data in larger and other population groups are needed to confirm this study. PMID- 17125003 TI - Health and nutrition survey of tsunami victims in Phang-Nga Province, Thailand. AB - The post-tsunami health and nutritional statuses of survivors were surveyed three months after the disaster struck. Non-participant observations and questionnaires were used to study the effects of the disaster on their lifestyles and health while residing in temporary shelters provided by the government and private donors. Anthropometrics were measured and dietary surveys conducted to elicit nutritional status. Our findings indicated good management of drinking water in the temporary shelters. Toilet construction and water supply were adequate, but wastewater and sewage systems were poorly managed. The study group still suffered from injuries after the disaster, and complained of back pain, stress, and sleep disorders. Most in the study group had unsatisfactory health behaviors, and obesity was an increasing problem among female participants. PMID- 17125004 TI - Barakol contents in fresh and cooked Senna siamea leaves. AB - Senna siamea (Lam.) Irwin and Barneby is a medicinal plant popularly used in Thailand. Young leaves and/or young flowers of this plant have been consumed by Thai people as a Khi Lek curry for a long time. The fresh young leaves and flowers are boiled with water 2-3 times to get rid of the bitterness and the boiled mush is used for curry cooking. Barakol, a major constituent of Senna siamea leaves was analyzed for its content in the fresh young leaves, the boiled leaves and the boiled filtrates by a high-performance thin-layer chromatographic method. Fresh young leaves of S. siamea contained 0.4035% w/w barakol. The amount of barakol in the first and second boiled filtrates were 0.2052 and 0.1079% fresh weight, while the first and second boiled leaves samples were 0.1408 and 0.0414% fresh weight, respectively. The results show the process of preparation of Khi Lek curry by boiling S. siamea young leaves twice with water reduced barakol content up to 90% and the content of barakol in boiled leaves used for curry has much less tendency to cause liver toxicity. This may explain the reason why Thai Khi Lek curry has not caused hepatotoxicity, unlike S. siamea leaves consumed as a powdered capsule. PMID- 17125005 TI - Incidence of enteric bacterial pathogens in water found at the bottom of commercial freezers in calabar, southeastern Nigeria. AB - Bacteriological analysis of water that accumulates at the bottom of freezers in restaurants when the power was cut in Calabar, southeastern Nigeria, was carried out using standard procedures. Mean heterotrophic bacterial counts and Escherichia coli counts ranged from 3.1 +/- 0.02 to 7.1 +/- 0.30 x 10(4) cfu/ml and 0.2 +/- 0.10 to 0.6 +/- 0.50 x 10(4) cfu/ml, respectively, indicating heavy bacterial contamination whose source was mostly fecal. There was no significant difference (p > 0.05, 0.01) in bacterial counts between freezers. Some biochemically identified enteric bacterial pathogens were Salmonella typhi, Shigella sp, enteropathogenic E. coli, Yersinia sp, Klebsiella pneumoniae, Vibrio cholerae O1 and Vibrio parahaemolyticus. This reveals that the hygienic quality of the food items stored in the freezers and the hygienic status of the restaurants are in doubt. Infection could be going on unnoticed and thus endemicity maintained in the area. The pathogens showed alarming antibiotic resistance. The water in the freezers was a "soup" in which different species of the enteric pathogens were close to each other and could transfer drug resistance among themselves. Public health education of restaurant operators in southeastern Nigeria is recommended. PMID- 17125006 TI - A modified dip-slide test for microbiological risk in caries assessment. AB - This study was undertaken to evaluate the validity of a new modified dip-slide test kit for the estimation of salivary mutans streptococci, lactobacilli and Candida levels by comparison with the results obtained from conventional agar plate counts. Five milliliters of paraffin-stimulated saliva sample was collected from 118 6-12-year old children attending 4 primary schools in Thailand. Saliva was poured over a 3-compartment slide containing Mitis-Salivarius Bacitracin agar, Rogosa agar and Sabouraud dextrose agar. A slide was incubated in a 5% CO2 incubator for 48 hours. The results of the modified test kit were compared with those from the conventional saliva sampling with dilution method. The salivary mutans streptococci, lactobacilli and Candida counts obtained from the conventional methods were significantly correlated (p < 0.0001) with the modified dip-slide estimates of these organisms. (Kendall Tau = 0.71, 0.58 and 0.76, respectively). The correlation between a modified dip-slide test kit and the conventional method indicates that this new test kit is suitable for the screening of salivary mutans streptococci, lactobacilli and Candida levels in patients. The test is simple and would be useful for the early selection of patients for dental examination. In addition, it would be a valuable educational aid for the motivation and dietary counseling among children. PMID- 17125007 TI - Estimation of non-fatal road traffic injuries in Thai Nguyen, Vietnam using capture-recapture method. AB - Road traffic injuries (RTIs) are increasing in developing countries where accurate routine data are usually not available. Although a capture-recapture technique has increasingly been employed in studies of human populations to provide reliable estimates of the magnitude of problems, it has rarely been used in road traffic injury research. We applied two sample capture-recapture methods using hospital and traffic police records to estimate non-fatal road traffic injuries in Thai Nguyen City during the years 2000-2004. We generated a conservative adjusted estimate of non-fatal RTIs using data from the two sources matched by name, surname, sex of victims and at least one of the other matching variables, of age, address of victim and date of injuries. We then compared the estimated rates with those reported based on police and hospital data. The results show that during years 2000-2004, the police reported 1,373 non-fatal RTIs, while hospital records revealed 6,069 non-fatal RTIs. Most reported victims on both hospital and police reports were males (67.3 % and 74.4%, respectively). More than half the victims on both hospital and police reports were drivers (77.5% and 66.1%, respectively) or pedestrians (10.6% and 7.1%, respectively). Youth and young adults (ages 15-34) constituted the majority of the victims on the hospital and police reports (52.8% and 63.7%, respectively). The capture recapture analysis estimated that 11,140 (95% CI: 10,626-11,654) subjects were involved in RTIs during the study period. In comparison to the estimated figure, official sources accounted for only 21.9 to 60.1% of total non-fatal RTIs. Estimated rates of non-fatal RTIs were 105.5 injuries/10,000 population per year and 393 injuries/10,000 vehicles. Given the fact that under reporting of RTIs has been a major limitation of routine official data sets in developing countries, we suggested the capture-recapture method be used as a tool to provide affordable and reliable estimates of RTIs in resource-poor countries. PMID- 17125008 TI - Reliability and construct validity of the Malay version of the Job Content Questionnaire (JCQ). AB - The JCQ has been shown to be a valid and reliable instrument to assess job stress in many occupational settings worldwide. In Malaysia, both the English and validated Malay versions have been employed in studies of medical professionals and laboratory technicians, respectively. The present study assessed the reliability and construct validity of the Malay version of the JCQ among automotive workers in Malaysia. Fifty workers of a major automotive manufacturer in Kota Bharu, Kelantan consented to participate in the study and were administered the Malay version of the JCQ. Translation (English-Malay) and back translation (Malay-English) of the JCQ was made to ensure the face validity of the questionnaire. Reliability was determined using Cronbach's alpha for internal consistency, whilst construct validity was assessed using exploratory factor analysis (principal component with varimax rotation). The results indicate that the Cronbach's alpha coefficients were acceptable for decision latitude (job control or decision authority) (0.74) and social support (0.79); however, it was slightly lower for psychological job demand (0.61). Exploratory factor analysis showed 3 meaningful common factors that could explain the 3 theoretical dimensions or constructs of Karasek's demand-control-social support model. In conclusion, the results of the validation study suggested that the JCQ scales are reliable and valid for assessing job stress in a population working in the automotive industry. Further analyses are necessary to evaluate the stability and concurrent validity of the JCQ. PMID- 17125009 TI - Benchmarks of Fairness for health care reform in Thailand--combining evidence with opinion of the civic group. AB - The concept of the Benchmarks of Fairness was tried in Thailand before the big reform of universal coverage policy in 2001. The first phase of the Benchmarks in 1999 involved the analysis of the national health reform proposal as well as the analysis of ongoing field trials of health reforms in two provinces. Though the participants were predominately health personnel, the results suggested the power of combining qualitative viewpoints of participants with the quantitative indicators within the province to move health reforms to more equitable, more efficient and more democratic directions. The second phase of the Benchmarks of Fairness, therefore, tested the possibility of involving wider participation of the civic groups related, and not-related to health, in assessing their provincial health system. The health achievements of the provinces a measured by 81 indicators, in the 9 benchmarks, were provided to the civic groups before focus group discussions in 10 selected provinces to facilitate discussions based on evidence. More qualitative data were obtained from the discussions as well as their judgements on the fairness of their provincial health system. Having completed this second phase, it was recommended that the benchmarks tool could be further endorsed as the basis for monitoring the progress of health reform by province and the effect of health care decentralization. To accomplish this monitoring, the civic groups should have continuous access to evidence, in line with the benchmarks, and they should be provided with the opportunity to express their views, which is helpful in monitoring fairness in the long run. PMID- 17125010 TI - [Prevention of risks and professional liability in anesthesiology]. PMID- 17125011 TI - [Cell saver devices in coronary revascularization surgery without extracorporeal circulation reduce transfusion requirements]. AB - OBJECTIVES: To analyze the effectiveness of a cell saver device in reducing transfusion requirements in patients undergoing off-pump coronary artery bypass surgery. PATIENTS AND METHODS: Fifty-six consecutive ASA class 4-5 patients who underwent coronary surgery without extracorporeal circulation in our cardiac surgery department between June 2004 and January 2005 were included in this retrospective study; the series comprised 28 patients who received conventional management (control group) without use of the cell saver device and 28 who received cell saver treatment. Variables analyzed were preoperative and discharge hemoglobin levels and hematocrit values, age, weight, height, ejection fraction, packed red blood cells transfused, exitus, and adverse events. RESULTS: The groups were similar with respect to preoperative characteristics. Fewer patients in the cell saver group required transfusions (6 vs 18 in the control group; relative risk 0.33, 95% confidence interval, 0.16-0.71). The mean amount of packed red cells transfused was greater in the control group than in the cell saver group (2.5 L vs 1.2 L, P = 0.03). No deaths or adverse events occurred in either group. CONCLUSIONS: The routine use of a cell saver device during off-pump coronary artery bypass surgery reduces the need for postoperative transfusions and is not associated with adverse events. Cell saver devices should be used routinely, especially in situations where the ability to provide blood transfusions may be compromised. PMID- 17125012 TI - [Application of a critical incident reporting and analysis system in an anesthesiology department]. AB - OBJECTIVES: To ascertain the changes in anesthesia-related morbidity and mortality after application of a scheme for reporting critical incidents and to assess the effect of implementing preventive measures against the detected errors. PATIENTS AND METHODS: We defined a critical incident to be any situation in which the margin of safety for the patient was reduced or might have been reduced. We analyzed data from the period between January 1999 and December 2004. RESULTS: The number of critical incidents was 547 (0.79% of 68627 anesthetic procedures). Human error was identified in 279 incidents (51%). The most frequent factors underlying errors were wrong diagnosis of the situation, communication problems, and failure to check equipment and drugs. The patient suffered no adverse effect in 81.8% of the incidents; 78.9% were considered preventable. Introducing an equipment checklist before anesthesia reduced the number of incidents from 90 events in 21809 cases in 31 months to 34 events out of 22064 cases in 29 months; chi2 test, P < 0.05; odds ratio (OR), 2.68; 95% confidence interval (CI), 1.80-3.98). Labeling syringes reduced errors in the administration of medications from 45 errors in 21 809 cases in 31 months to 27 in 22064 cases in 29 months; chi2, P < 0.05; OR, 1.68; 95% CI, 1.04-2.72. CONCLUSIONS: Corrective measures were adopted as a result of the incident reporting scheme. Some of the measures led to a statistically significant reduction in equipment and drug administration errors. PMID- 17125013 TI - [Autoradiography of central benzodiazepine receptors in the hippocampus during morphine clearance in surgically manipulated rats]. AB - OBJECTIVES: To determine whether the stress caused by minimally invasive surgery alters benzodiazepine receptor subtypes in different areas of the hippocampus and to characterize the role that loss of morphine tolerance plays in surgical stress. MATERIAL AND METHODS: Minimally invasive surgical manipulation consisted of implantation of subcutaneous pellets under halothane anesthesia. The experiments were performed in 9 groups of male rats: naive rats (no manipulation), 4 sham groups (sacrificed 6, 12, 24, and 36 hours after removal of sham pellets), and 4 morphine groups (sacrificed 6, 12, 24, and 36 hours after removal of morphine releasing pellets). Receptor autoradiography was performed with receptor saturation binding studies. The benzodiazepine receptors were [3H]flunitrazepam labeled; zolpidem binding was used to characterize receptor subtypes. RESULTS: The surgically manipulated groups, whether treated with morphine or not, displayed increased benzodiazepine receptor density in comparison with naive animals for all receptor subtypes. No significant differences in radioligand affinity were observed for the various receptor subtypes under any of the experimental conditions. Benzodiazepine receptor uptake did not differ significantly between morphine treated and untreated animals. CONCLUSIONS: Surgical stress increases benzodiazepine receptor density but does not alter affinity. Loss of tolerance to morphine does not seem to play an important role in surgically caused stress. PMID- 17125014 TI - [Incident reporting systems and patient safety in anesthesia]. AB - Incident reporting schemes collect information on adverse events, errors, complications, or problems with the aim of analyzing their causes and suggesting changes to prevent recurrence. Such schemes are currently part of clinical safety programs in various countries. Although the ideal form for a reporting system is debated, an essential part of its success will be the establishment of a culture of safety within an organization. The underlying assumption is that even though errors are an inherent part of a process that relies on human beings, they are nearly always favored by a chain of system failures. Therefore, reporting is intended to stimulate a culture of learning rather than assigning blame. The main limitations of such schemes are under reporting, the use of different terms and concepts, the lack of resources for research and development, and the scarcity or lack of legislation to guarantee the proper use of information without legal consequences. PMID- 17125015 TI - [Anesthetic management for left ventricular epicardial lead implantation with minimally invasive thoracoscopic guidance]. AB - Medical treatment for left ventricular (LV) systolic dysfunction and congestive heart failure has improved quality of life for patients but mortality rates have remained unaffected. For a subgroup of such patients with interventricular conduction delays and ventricular contraction dyssynchrony, cardiac resynchronization by placement of a LV epicardial lead is a new approach to management. We report 3 cases in which such electrodes were implanted under the guidance of minimally invasive thoracoscopy. In the first 2 cases it was decided to place the LV electrode using thoracoscopic guidance because of complications or technical difficulties in the percutaneous procedure. In the third case thoracoscopy was used because of deterioration of the patient's condition after implantation of a double-chamber pacemaker and shifting of the lead to the right ventricle. The literature on the anesthetic management of such patients is scarce. Although sedation with general anesthesia and single-lung ventilation is indicated for percutaneous procedures, that technique also proved adequate for the thoracoscopic procedures. PMID- 17125016 TI - [Systemic toxicity caused by local anesthetics after peripheral nerve blocks]. AB - Peripheral nerve blocks have aroused increasing interest in recent years, leading to a rise in the rate of complications. At the same time noteworthy technical advances have been made in areas such as nerve stimulation and ultrasound imaging, and local anesthetics have become safer. Nevertheless, the risk of anesthetic-related systemic toxicity, which manifests with neurological symptoms that tend to be forerunners of cardiovascular ones, can not be ignored. We report 2 cases of systemic toxicity due to the use of a mixture of local anesthetics during nerve blocks for outpatient surgery. PMID- 17125017 TI - [Complex regional pain syndrome type 2 after radial artery catheterization: a case report]. AB - Complex regional pain syndrome has multiple causes. The clinical picture includes pain that can be debilitating, along with vascular and motor abnormalities, changes in sweating, delayed recovery, eating disorders, and occasionally psychological changes. Treatment is complex and should be started early if symptoms are to be reversed. We report the case of a man who developed complex regional pain syndrome type 2 in his left arm after surgery with extracorporeal circulation to repair an interatrial septal defect. The clinical picture was believed to have been triggered by catheterization of the radial artery. PMID- 17125018 TI - [Diagnostic delay in neuroleptic malignant syndrome]. PMID- 17125019 TI - [Anaphylactic reaction to latex after surgery and release of the ischemic cuff]. PMID- 17125020 TI - [Remifentanil: bolus doses to induce apnea and bradycardia for coronary arteriography in a pediatric patient]. PMID- 17125021 TI - The combined spinal-epidural analgesia for labor pain. PMID- 17125022 TI - [On amniotic fluid embolism]. PMID- 17125023 TI - [Catheter knotting during continuous brachial plexus analgesia by the paraescalene approach]. PMID- 17125024 TI - [Mastocytosis and anesthesia: a case report]. PMID- 17125025 TI - [Newly located Spanish doctoral theses and expert opinion papers on anesthesia]. PMID- 17125026 TI - [Megacolon]. PMID- 17125027 TI - [Occupational diseases in Poland, 2005]. AB - BACKGROUND: The aim of this paper was to present basic statistical data on occupational diseases diagnosed and certified in 2005. MATERIALS AND METHODS: This work is based on the data compiled from "Occupational Disease Reporting Forms" received by the Central Register of Occupational Diseases in 2005. The data comprised information on individual diseases, gender and age of patients, and duration of occupational exposure to harmful agents responsible for the development of specified pathologies. These data were further classified by sectors of the national economy and voivodships (provinces). The incidence was specified in terms of the number of cases per 100,000 paid employees and per 100,000 employed persons. RESULTS: In Poland, the number of occupational diseases diagnosed in 2005 accounted for 3249 cases. The incidence rate was 34.8 cases per 100,000 paid employees. The highest incidence rates were noted for: chronic disorders of the voice organ (7.3 per 100,000), pneumoconioses (7.2), infectious and parasitic diseases (6.6) and permanent bilateral hearing loss (3.6). As much as 73.0% of patients affected by occupational diseases had been exposed to harmful agents for more than 20 years. In the industrial sectors of the national economy, the highest incidence was noted among workers employed in the mining industry (281.7 cases per 100,000 paid employees), particularly in coal mining (325.2). The highest incidence rates were recorded in the Podlaskie (87.3), Lubelskie (82.1), and Silesian (66.3) voivodships. CONCLUSION The incidence of occupational diseases in Poland continues to abate. The downward trend is primarily due to a lower incidence of chronic diseases of the voice organ and hearing loss. However, an increased number of cases of infectious and parasitic diseases, mainly borreliosis, was revealed. PMID- 17125028 TI - [Tuberculosis as an occupational disease in the province of Wielkopolska]. AB - BACKGROUND: The incidence of tuberculosis (TB) in the general population of Poland is still quite high as compared to the "old" Fifteen of the European Union. In Poland, occupational TB has recently been diagnosed almost entirely among medical personnel and social workers. The aim of the study was to analyze TB epidemiology in the province of Wielkopolska, taking account of the largest pulmonological hospital in this region. MATERIAL AND METHODS: The analysis was based on the data contained in the occupational disease certification forms collected by the State Sanitary Inspection for the province of Wielkopolska as well as on the data on TB incidence and employment of workers derived from the largest hospital of pulmonological and phthisiological profile. RESULTS: In the Wielkopolska region, 7 cases (17.5%) of occupational TB in men and 33 cases (82.5%) in women were found in 1999-2003. In those years, TB incidence was not significantly higher among physicians (18.6/100,000/year) and nurses (26.9/100,000/year) than in the general population (20.4/100,000/year) of this region. In the study population, age was a significant factor influencing the incidence of occupational TB. Nurses were the occupational group that was at particular high risk of TB at the young age (mean, 34.4 +/- 6.5 years). Physicians fell ill much later--the mean age of diagnosing TB in this group was 53.8 +/- 12.1 years. In the hospital under study (median number of employees in the years 1997-2003 was 419 persons), the median morbidity from TB was 4.7 cases/1000 employees/year; 7.1/1000 physicians/year; and 3.9/nurses/year in 1997 2003. CONCLUSIONS: The results of this analysis confirmed the influence of age and character of the medical institution on TB epidemiology among medical personnel. In this paper the problems of epidemiology of certified cases of occupational tuberculosis are also discussed. PMID- 17125029 TI - [The state of teeth and low salivary secretion in miners working at different workposts and depths]. AB - BACKGROUND: The aim of this work was to find out whether a relationship between the workpost in the mine and the state of teeth and salivary secretion does exist. It was investigated whether different environmental conditions find their reflection in miners' dentition, and also whether excessive dustiness can be responsible for low salivary secretion. MATERIALS AND METHODS: Four groups of miners were analyzed: those working in coal face and walls, and those working at depths of 525 and 1030 m. Martin's and DMFT indices were used to assess the condition of teeth. RESULTS: The lowest value of DMFT coefficient (18.2) was found in miners working in walls and the highest (19.7) in miners working in coal faces; the values of Martin's coefficient were 16.68 and 17.46, respectively. The difference between values for miners working at depths of 525 and 1030 was insignificant (0.1). Arithmetical means for saliva investigation were very similar in all groups. CONCLUSIONS: There were no essential differences in the dentition of the miners working at different workposts. The relationship between excessive dustiness, reduced saliva secretion and increased DMFT coefficients was ascertained. PMID- 17125030 TI - [Contact allergy to preservatives contained in cosmetics]. AB - BACKGROUND: The aim of this study was to assay the type of allergy to preservatives contained in cosmetics and to assess the usefulness of the composition of allergens included in a standard series for the diagnosis of occupational contact allergy used to date. MATERIALS AND METHODS: The frequency of contact allergy to the standard series of preservatives (thimerosal, Euxyl K 400, formaldehyde, Kathon CG, Quaternium 15, parabens) was assayed in a group of 1937 subsequent patients referred to the Nofer Institute of Occupational Medicine in L6di and examined in the years 2000-2005. The frequency and type of allergy to preservatives of a cosmetic series in a group of 113 patients with poor tolerance of cosmetics were also investigated. RESULTS: Allergy to thimerosal was found in 11.8% of patients tested with a standard series; to formaldehyde in 4.9%; and to Euxyl K 400 in 3.7%. Quaternium and parabens were less allergenic (0.8 and 0.3%, respectively). In the group of 113 patients subjected to patch test with a cosmetic series, allergy, i.e. at least one positive patch test, was observed in 49 (43.4%) patients. Of the 27 preservatives contained in cosmetics, 16 induced positive reaction to chemical compounds. Euxyl K 400 proved to be the basic allergenic preservative of this series, and induced allergy in 21 (18.6%) patients, whereas 17.7% of patients reacted to thimerosal. Only allergy to thimerosal applied to 8 persons, and 7 of them showed inflammatory lesions only on the face. Less allergic biocides were cocamidopropyl betaine (7.1%), Kathon CG (7.1%), Bronopol (5.3%), Germall II (4.4%), triethanolamine (3.5%), Germall 115 (2.6%), DMDM hydantoin (2.6%), Grotan BK (1.8%), sodium-2-pyridinethiol-1-oxide (1.8%), clioquinol (0.9%), Quaternium 15 (0.9%), and dimethylaminopropylamine (0.9%). CONCLUSIONS: The results of our study confirmed observations of other authors that allergy to preservatives present in numerous industrial products, especially Euxyl K 400, is still a growing problem. Like many other researchers, we are of the opinion that this allergen should be included in a standard series for the diagnosis of all patients with suspected allergic dermatitis, including that of occupational origin. PMID- 17125031 TI - [Diagnostic x-ray examinations in Poland in 2004 in view of the population exposure: the assessment of structure and trends]. AB - INTRODUCTION: The paper presents the results of the Polish national survey of medical x-ray examinations performed in 2004. MATERIALS AND METHODS: A questionnaire was used to collect the data on the number and types of x-ray examinations. The data covered most of x-ray laboratories in Poland, being under the dosimetric surveillance performed by the Radiation Protection Department at the Nofer Institute of Occupational Medicine in Lodz. RESULTS: The number of x ray procedures in Poland in the year 2004 accounted for 32.4 million examinations, i.e. 830 procedures per 1000 inhabitants; the latter figure includes 65 specialist procedures and 61 dental examinations per 1000 of inhabitants, each. They were performed in over 3000 x-ray laboratories, including operational theatres and dental units. The majority of patients were at the age of 40-60 years and women made over 50% of all those examined. CONCLUSIONS. Owing to the modification of the heath care organisation, the profile of examinations performed in x-ray laboratories has changed. Nevertheless, conventional x-ray examinations are still in a majority, among which radiographies of chest, spine and limbs are most frequently performed. It should be emphasised that the number of the hip joint radiographies in infants and small children have decreased and practically photofluorographic examinations have been eliminated from the diagnostic practice. The number of specialist procedures has evidently increased, and thus the frequency of these procedures in Poland and other European countries is comparable. PMID- 17125032 TI - [Weighing precision of filters used in dust measurements: repeatability and reproducibility]. AB - BACKGROUND: Measurements of inhaled and respirable dust concentrations in the work environment are most frequently performed by employing the method of the air filtration with analytical filters placed in measuring heads, which is followed by the mass determination of the dust precipitated on the filter respective to the volume of the filtrated air. A twofold weighing of filters is thus the basic measurement technique used to determine the dust mass. It should be emphasized that the weighing precision decides about the precision of measured inhaled and respirable dust concentrations. MATERIAL AND METHODS: The weighing precision of FiPro polypropylene filters, commonly used in industrial dusts measurements in Poland, was assessed. Ten filters of phi 25 mm and 37 mm were placed in open containers and desiccators with constant air humidity of 5% in weighing room. After 24 h, filters were weighed five times at one-week intervals. Weighing was performed by two persons on two balances (0.01 mg); 21 accredited laboratories participated in inter-laboratory comparisons; each of them weighed twice 5 filters of phi 25 mm and 37 mm kept in desiccators for 24 and 48 h. RESULTS: Standard deviation of repeatability was calculated--mass Sr of filters of phi 25 mm equal to 0.016 mg was 1.4 times lower than mass Sr of filters kept in an open container, whereas mass Sr of filters of phi 37 mm from desiccator, also equal to 0.016 mg, was 1.6 times lower than mass Sr of filters from an open container. Mean standard deviations of reproducibility, S(R), calculated on the basis of the results of inter-laboratory studies (after rejecting significantly departing values) were 0.024 mg and 0.022 mg for filters of phi 25 mm after keeping them in desiccator for 24 and 48 h, whereas these values accounted for 0.058 mg and 0.033 for filters of phi 37 mm. CONCLUSIONS: The results of the study in conditions of repeatability confirm high mass stability of FiPro polypropylene filters. It may be elucidated from inter-laboratory comparisons that keeping FiPro filters in an desiccator for at least 48 h before and after collection of air samples will ensure the highest precision of their mass determination, and thus the highest precision of determinations of inhaled and respirable dust concentrations. PMID- 17125033 TI - [Influence of prolonged sedentary work on the development of lower limbs edema and methods of its prevention]. AB - Prolonged sitting may lead to numerous complications of general nature such as fatigue or discomfort, but also to diseases of the musculosceletal system (the spine and upper limbs), chronic veins insufficiency, deep veins thrombosis as well as obesity with its further, mostly cardiovascular consequences. Due to the dynamic growth of new workplaces associated with the introduction of modern technologies, the frequency of these health problems has increased over the recent years. This paper presents an overview of complications of long-term work performed in a sedentary position on the functioning of the lower limbs venous system. The etiology of lower limbs edema, diagnostic methods, and proposed preventative measures to eliminate or reduce swelling of the legs are presented, including optimization of workplaces, organizational interventions, and promotion of the health culture. PMID- 17125034 TI - [Trials of casual treatment of silicosis]. AB - Silica-induced lung injury and the development of silicosis is one of the major occupational diseases. Accumulation and deposition of respirable dust containing silica mineral particles in the lung produces chronic lung disease characterized by granulomatous and fibrotic lesions. Knowledge of precise mechanisms, which induce this process is still limited, hence problems faced in the treatment of silicosis, especially the casual one. This article describes various trials of casual silicosis treatment with tetrandrine (Tet), isolated from the root of Stephania tetrandra, tumor necrosis factor (TNF) antagonists, polyvinyl-pyridine N-oxide (PVNO), aluminum compounds, corticosteroids or bronchoalveolar lavage (BAL). The existing methods are not sufficient, which warrants further investigations. At present, prevention of the disease and treatment of its complications are most important. PMID- 17125035 TI - [The effect of occupational and non-occupational psychosocial stress on the course of pregnancy and its outcome]. AB - The effect of stressful professional work on the course of pregnancy and its outcome is for various reasons an important issue. The aim of this work was to compare and evaluate studies carried out in different countries all over the world on the risk of preterm delivery (PTD) and fetal hypotrophia in infants whose mothers were occupationally exposed to psychosocial stress during gestation. An analysis of available literature confirms a significantly higher PTD odds ratio among pregnant women working in adverse psychosocial conditions. Low birth weight (LBW) in newborns of this group of mothers occurs more frequently than in the general population. PMID- 17125036 TI - [Possible consequences of acetylcholinesterase inhibition in organophosphate poisoning. Discussion continued]. AB - Numerous toxicologists still believe that the only function of acetylcholinesterase (AChE) is to catalyse the hydrolysis of acetylcholine (ACh), and that the toxicity of organophosphorous pesticides (OP) results from their ability to switch this function off. This viewpoint, however, requires revision in the light of the recent findings concerning the AChE structure and function as well as in view of the regulation of the AChE protein synthesis in conditions of stress. There is now no doubt that apart from catalysing the ACh hydrolysis, AChE performs also nonenzymatic functions, trophic (e.g., stimulation of neuritogenesis and remodeling) and neuromodulatory (promotion of long-term functional changes in the central nervous system). Binding to OP does not interfere with its nonenzymatic functions, but it stimulates AChE mRNA transcription and overproduction of the AChE protein. This results in overexpression of nonenzymatic AChE functions. It is likely that this overexpression is responsible for at least some of the persistent alterations in the CNS functions which may develop after an OP exposure. PMID- 17125037 TI - [Medycyna Pracy: the scopus-based analysis of citations]. AB - Medycyna Pracy, a Polish bimonthly published since 1950, forms a long-standing documentation of studies carried out in the area of workers' health protection. The journal is primarily addressed to occupational health physicians and work hygiene specialists in Poland. It is indexed by numerous foreign information services (e.g., MEDLINE, EMBASE, BIOSIS PREVIEWS, BIOLOGICAL ABSTRACTS, SCOPUS) and thus promotes Polish research in occupational medicine throughout the world. The quantitative analysis for the years 1996-2005, grounded on the SCOPUS database, presents an average number of citations concerning a given volume, frequency of citations, articles most frequently cited, and countries, in which articles published in Medycyna Pracy have been referred to. A growing number of citations observed in the recent years signify the importance of issues investigated and discussed in the journal as well as its role in the world-wide circulation of scientific information. PMID- 17125038 TI - [The verified Polish regulations on the maximum admissible intensities (MAI) of electromagnetic fields]. PMID- 17125039 TI - [The Monaldi Archives: A new instrument of communication and information in Rehabilitation Cardiology]. PMID- 17125040 TI - The use of "LAST" operation in high risk patient. AB - BACKGROUND: The LAST operation, in spite of few drawbacks, represents a good option for single Left Anterior Descending (LAD) revascularization. This procedure does not allow multivessel revascularization, where hybrid procedure have been previous described. We report preliminary experience with the LAST operation performed in high risk patients. MATERIAL AND METHODS: From October 2004 to February 2005, 11 male high risk patients (mean age 74 +/- 8 years) underwent the LAST operation. Mean predicted death with EUROSCORE and Parsonnet score were 29% and 31% respectively. All patients had a proximal LAD lesion either not suitable for PTCA and multivessel coronary artery disease. The mean preoperative Ejection Fraction was 42 +/- 5% (27-55%). Four patients (36.4%) had previous surgical myocardial revascularization. An incision of about 6 cm was made in the appropriate intercostal space and the LIMA (Left Internal Mammary Artery) was harvested using a special costal retractor. After heparin administration the LIMA is distally divided to check the adequacy of the blood flow. Following the insertion of a temporary intracoronary shunt, the LIMA was LAD anastomosis was carried out with a continuous 8-0 polypropylene suture. RESULTS: No hospital or late mortality was observed. Uneventful conversion to midline sternotomy was necessary in one patient with low value of mammary flow. All patients were discharged within the first postoperative week. CONCLUSIONS: The use of the LAST operation enhances the role of minimally invasive surgery also in high risk patients who need coronary revascularization. PMID- 17125041 TI - Efficacy of telecardiology in improving the results of cardiac rehabilitation after acute myocardial infarction. AB - BACKGROUND: This study was addressed to verify if Telecardiology (TC) improves the results of Cardiac Rehabilitation in patients following a home-based Cardiac Rehabilitation Program (CRP) after acute myocardial infarction (AMI). MATERIALS AND METHODS: We studied three groups of patients after AMI: Group A (control group): 15 patients, who followed a standard in-hospital CRP of 3 weekly sessions of 2 months duration; Group B (study group): 15 patients, who were enrolled in a home-based CRP of similar duration and were monitored by TC with the aid of an ecg-device (Sorin Life Watch CG 6106); Group C (second control group): 15 patients, who followed a home-based CRP without ecg-monitoring by TC. All patients performed a symptom-limited exercise testing at the beginning of the CRP. Psychometric data (STAI-Y1, STAI-Y2, BDI) were also evaluated. At the end of the CRP all patients underwent repeated exercise testing and psychometric evaluation. RESULTS: TC applied to the home-based CRP was associated with a good compliance to the program. Compared to Group C, in Group B we observed an increase of maximal heart rate, exercise duration, maximal work-load, and an improvement of anxiety, a trend to reduction of depression, and an improvement of quality of life. These results were very similar to Group A patients following a hospital-based CRP. CONCLUSIONS: TC improves compliance, functional capacity and psychological profile of patients undergoing a home-based CRP, compared to patients enrolled in a home-based CRP without ecg-monitoring by Telecardiology. PMID- 17125042 TI - [Acute myocardial infarction in the elderly. A case-control study with a younger population and review of literature]. AB - BACKGROUND: ST elevation myocardial infarction (STEMI) in old and old-old patients presents several peculiarities in natural history, delay of hospitalization and response to treatment. Aim of this retrospective case control study was to determine presentation, complications and management of elderly patients with STEMI compared to a younger population. METHODS: 462 patients (205 M and 257 F) aged > or =75 years, hospitalized in CCU between 1999 and 2003 for STEMI, were evaluated. The control group consisted of 490 consecutive patients (268 M and 222 F) aged 50-70 years. Attention was focused on clinical presentation, complications, management and outcome in elderly compared with younger patients. RESULTS: The mean interval between the onset of symptoms and the arrive in CCU was of 9 hour in the elderly compared to 4,5 hour in the control. Chest pain was less frequent (50% vs 90%) in the elderly; the prevalence of dyspnoea and neurological symptoms was higher in patients >75 years (30% vs. 15% and 25% vs. 10%). In the elderly, previous angina and AMI, cerebral and peripheral vascular diseases, peripheral and renal failure were frequent. Early severe complications prevailed in the elderly. Thrombolysis was performed only in 39% of the elderly compared to 65% of the control. Significantly higher was cerebral haemorrhage after thrombolysis (4.9% vs. 1.8%). Comparable were the mayor extra cranial bleedings. Primary or facilitated PTCA was performed in few patients in the last year. Two weeks mortality was 20%, compared to 6.5% in the control group. CONCLUSION: The patients >75 years with STEMI were hospitalized later, had atypical presentation with less chest pain and more cardiac failure, were less likely to receive thrombolysis, had more complications and more cerebral bleedings. Elderly had more associated diseases and in-hospital mortality was higher. PMID- 17125043 TI - [From risk charts to guidelines: tools for evaluation and management of cardiovascular risk]. AB - Despite the wide improvement of diagnostic techniques and the introduction of effective pharmacological and instrumental therapeutic strategies aimed to the treatment of cardiovascular diseases, their incidence and lethality are still elevated, with economic implications increasingly less sustainable by the public medical systems. The modern practice of cardiovascular prevention requires, thus, that diagnostic and therapeutic interventions, both at population level and on the single patient, should be more and more precise, effective, and appropriate. From this point of view, a correct global cardiovascular risk stratification assumes a preponderant relevance, in order to allow an adequate therapeutical response. For this purpose several work instruments, as risk charts and guidelines, namely dedicated to arterial hypertension and dyslipidemias, were developed and offered to clinicians interested in cardiovascular prevention. The aim of this review is to illustrate, in synthesis, those instruments, aiming to facilitate their implementation, thus reducing the actual gap between theoretical indications and the real world. PMID- 17125044 TI - Trends in cardiovascular rehabilitation. PMID- 17125045 TI - [Polycystic ovary syndrome: an example of obesity-related cardiovascular complication affecting young women]. AB - Polycystic ovary syndrome (PCOS) is a good example of obesity-related cardiovascular complication affecting young women. PCOS is not only considered a reproductive problem but rather represents a complex endocrine, multifaceted syndrome with important health implications. Several evidences suggest an increased cardiovascular risk of cardiovascular disease associated with this syndrome, characterized by an impairment of heart structure and function, endothelial dysfunction and lipid abnormalities. All these features, probably linked to insulin-resistance, are often present in obese PCOS patients. Cardiovascular abnormalities represent important long-term sequelae of PCOS that need further investigations. PMID- 17125046 TI - Interventional strategies in early atherosclerosis. AB - Atherosclerosis remains clinically mute for a long time and frequently manifests itself with an acute cardiovascular event; therefore, the possibility to detect the disease in a subclinical phase and to reduce or reverse its progression is an issue of relevance. Non-invasive diagnostic procedures such as B-mode ultrasonography of carotid intima-media thickness (CIMT), electron beam computed tomography (EBCT) and magnetic resonance angiography (MRA) allow to identify atherosclerotic disease in its early phases, to evaluate the disease progression and monitor the effects of interventions. In recent years, several therapeutic strategies have been adopted over time to slow early atherosclerosis in asymptomatic individuals at intermediate/high cardiovascular risk. Prospective trials employing multifactorial non-pharmacological interventions (diet, exercise, smoking cessation) have demonstrated a favorable effect on progression of atherosclerosis. Hence lifestyle modification may be an effective therapeutic strategy to be adopted as a first step and a highly cost-effective intervention in a preclinical setting involving a large number of individuals. Drugs should be considered as a second step or should be associated to further reduce the risk in individuals at high probability of future events, who require more intensive interventions. Reducing low-density lipoprotein levels, blood pressure and platelet aggregation seems to be the most effective intervention in these subjects, whereas the treatment of emerging coronary risk factors, e.g. oxidative stress, inflammatory activation and infections has not produced the expected protective effect. PMID- 17125047 TI - [Heart failure in Eastern Veneto: prevalence, hospitalization rate, adherence to guidelines and social costs]. AB - Heart failure is a prominent problem of public health, requiring innovating methods of health services organization. Nevertheless, data are still not available on prevalence, hospitalization rate, adherence to Guidelines and social costs in the general Italian population. The necessity to identifying patients with heart failure derives from the efficacy of new therapeutic interventions in reducing morbidity and mortality. In this study we aimed to identify, in a subset of the Eastern Veneto population, patients with heart failure through a pharmacologic-epidemiologic survey. The study was divided in 5 phases: (1) identification of patients on furosemide in the year 2000 in the ASL 10 of Eastern Veneto general population, through an analysis of a specific pharmaceutic service database; (2) definition of the actual prevalence of heart failure in a casual sample of these patients, through data base belonging to general practitioners, cardiologists, or others. Diagnosis was based on the following criteria: (a) previous diagnosis of heart failure; (b) previous hospitalization for heart failure; (c) clinical evidence, with echocardiographic control in unclear cases; (3) survey of hospitalizations; (4) evaluation of adhesion to guidelines, through both databases and questionnaires; (5) analysis of the social costs of the disease, with a retrospective "bottom up" approach. From a total population of 198,000 subjects, we identified 4502 patients on furosemide. In a casual sample of 10,661 subjects we defined a prevalence of heart failure in Eastern Veneto of 1.1%, that increased to 7.1% in octagenarians. The prescription of life saving drugs was satisfactory, while rather poor was the indication to echocardiography and to cardiologic consultation. Hospitalization rate for DRG 127 was low: 2.1/1000 inhabitants/year in the general population and 12.5 /1000 inhabitants/year in patients >70 years of age. Yearly mortality was 10.3%. Social costs were elevated (15.394 Euros/patient/year), due to a relevant sanitary component (hospital 53%, drugs 28%) and particularly a to an indirect cost component. In conclusion, the assumption of furosemide lends itself as a good marker for identifying patients with heart failure. Patient identification is simple, cheap and cost-efficient, and can be easily reproduced in other regional areas. PMID- 17125048 TI - [National Guideline in Rehabilitation Cardiology and secondary prevention in cardiovascular diseases]. PMID- 17125049 TI - [First National Conference: "rehabilitative and preventive cardiology, in the era of interventional cardiology." Presentation of rationales and objectives .Naples, March 3-4, 2006]. PMID- 17125050 TI - [Cardiologic rehabilitation: the guideline and the Italian reality]. PMID- 17125051 TI - [The resources in Cardiac Rehabilitation ]. PMID- 17125052 TI - [How to optimize the treatment after acute phase: which programmes for which patients?]. PMID- 17125053 TI - [The necessity of multidisciplinary competence and the accessory value of Rehabilitative Cardiology in the cardiosurgery patient ]. PMID- 17125054 TI - [The role of Rehabilitation Cardiology in the programs of disease management in heart failure]. PMID- 17125055 TI - [Timing and modality of control after acute coronary syndrome treated with revascularization: some reflections]. PMID- 17125056 TI - [Round table. Can be shared and implemented the suggested guidelines in the real world? Which are the real obstacles in the development of rehabilitation prevention and how to overcome it? ]. PMID- 17125057 TI - Vagal complexity: substrate for body-mind connections? PMID- 17125058 TI - A response to Mravec and Hulin: does the vagus nerve constitute a self organization complexity or a "hidden network"? PMID- 17125059 TI - Commentary to the article Does vagus nerve constitute a self-organization complexity or a "hidden network"? written by Mravec B. and Hulin I. in BMJ 2006. PMID- 17125061 TI - Factors influencing graft potency in patients who underwent CABG for treatment of CAD. AB - OBJECTIVES: To identify factors that influence graft disease and native coronary arteries progression disease and prognostic implication of this process. BACKGROUND: Unsolved problem in CABG patients is progression of the disease in bypass grafts and native coronary arteries. METHODS: Data from 102 patients with CABG, who underwent re-coronarography, were analyzed: - Pre and post-operative variables: risk factors, clinical status, functional capacity, left ventricular parameters and angiographic status (before and after CABG). Proportional hazard regression model, was used, p<0.05 was considered statistically significant. RESULTS: Cardiac death, myocardial infarction and heart failure were more frequent in patients with graft occlusion, non-stable angina pectoris in non occlusive graft disease, which together with acute myocardial infarction was more often in patients with native coronary arteries progression disease. PCI was significantly more often performed on native coronary arteries. CONCLUSION: Graft disease and native coronary artery progression disease is a continuous process which can be slowed by aggressive risk factors reduction, medications, and PCI procedures. In contrary, it leads to unfavorable clinical outcome (Tab. 8, Fig. 6, Ref 19). PMID- 17125062 TI - Diagnostics of incomplete Brown-Sequard syndrome caused by meningococcal myelitis. AB - We are reviewing a 20-year old girl with the history of acute meningococcal sepsa with meningitis and thoracic myelitis resulting from an early embolia into the spinal cord, which is a very rare complication of meningococcemia. The ipsilateral loss of sensation indicates lesion of the posterior ascendent fiber tracts in the spinal cord, witout contralateral motoricity deficite, this indicating the incomplete Brown-Sequard syndrome. Somatosensory-evoked potentials (SSEP) of the n. tibialis show milder impediments of conductivity by the thoracic segment at left. Magnetic resonance (MRI) of the thoracic spine shows lesion of the posterior ascendent fiber tracts (Fig. 2, Ref. 17). PMID- 17125063 TI - Treatment of postpsychotic depression with sertraline in patients with schizophrenia. AB - Some authors treated a two groups of patients with postpsychotic depression in a groups of patients with schizophrenia. Sertraline proved better than imipramine in view of earlier onset of action and lower incidence, intensity and duration of adverse effects and lower risk of schizophrenie symtom recurrence (Ref 4). PMID- 17125064 TI - Diagnostic approach to hypercoagulable states. AB - The primary hypercoagulable states are inherited thrombotic disorders, resulting from mutations in genes encoding a plasma protein component of one of the coagulation mechanisms. The anticoagulant pathways most frequently involved include antithrombin III, protein C, and protein S deficiencies and activated protein C (APC) resistance. Around 80 % of all individuals with APC resistance carry a mutation of the factor V gene. Abnormalities in procoagulant pathways mostly concern elevated levels and/or function of procoagulant factors. Elevation in plasma prothrombin (FII) levels is associated with a FII gene mutation. Hyperhomocysteinemia as a risk factor for thrombosis is determined by genetic or dietary factors. The acquired or secondary hypercoagulable states consist of a heterogeneous group of disorders with an increased risk for developing thrombosis. Many underlying conditions (e.g., malignancies) may induce changes in the coagulation system. The risk of thrombosis is increased in individuals with antiphospholipid antibodies. They are found in about one-half of patients with systemic lupus erythematosus, but also in the course of infections, neoplastic diseases, and sometimes in apparently normal subjects. A definite molecular abnormality of hypercoagulable states can be identified in the special coagulation laboratory, using two types of molecular risk factors for thrombosis (genetic factors and abnormal laboratory phenotypes). Its recognition is useful for a prevention and/or therapy of thrombosis (Tab. 4, Ref. 10). PMID- 17125065 TI - Septic complications of acute pancreatitis. AB - Acute pancreatitis (AP) is a potentially lethal disease. There are numerous studies published on acute pancreatitis. This article presents the results of research of many scientists in the field of acute pancreatitis. The main aim of this article is to present the possible septic complications of acute pancreatitis, its diagnostic and treatment modalities. Early morbidity and mortality are the result of activation of mediators with failure of circulation and other organ systems. The overall mortality of patients with acute necrotising pancreatitis is in the range of 10-15 %. Secondary pancreatic infection and sepsis develop in 40-70 % of patients with 80 % mortality. Pancreatic infection is caused by bacterial contamination of pancreatic necrosis. Infection is usually recorded in the second week of the disease in 24 % and in 71 % during the fourth week of the disease. The incidence of secondary infection and sepsis correlates with the extent of pancreatic necrosis. The prevention of infection and sepsis by systemic administration of antibiotics is considered a principal step in the therapy of acute pancreatitis (Ref 62). PMID- 17125066 TI - Cardiovascular risk in patients with chronic kidney diseases: a time for new risk markers? AB - The number of patients with end-stage renal disease who need expensive dialysis treatment or transplantation is increasing all over the world. Therefore, new possibilities of prevention and treatment of chronic kidney diseases (CKD) are being looked for. However, patients with CKD represent a population not only at risk for progression to renal failure, but also at even greater risk for cardiovascular (CV) diseases. Extreme CV morbidity and mortality in dialysed patients is well-known, but recent studies stress a high CV risk even at early stage of CKD. Impaired renal function and/or albuminuria are now considered major CV risk factors. The reasons for this are not only a high prevalence of traditional CV risk factors in CKD patients or their insufficient treatment, but also the presence of non-traditional CV risk factors that could be specific for CKD. Some of these novel risk factors, particularly markers of inflammation, endothelial function, hemocoagulation and fibrinolysis are discussed, and their significance in the prediction of CV and renal outcome examined (Tab. 1, Ref 57). PMID- 17125067 TI - Penile block in children, our first experience. AB - Penile block has been in use in penile surgery as a technique for perioperative analgesia for 25 years. The authors reintroduced penile block on the Department of Paediatric Anaesthetics and Intensive Care at the University Children's Hospital in Bratislava, Slovakia in June 2004 after 8 years. This prospective open observational study presents their experience with penile block for elective penile surgery in 96 paediatric patients (Tab. 3, Ref. 14). PMID- 17125068 TI - A review of ano-rectal disorders and their treatment. AB - This article describes the techniques of diagnosis and office-based interventions. From this review one can presume that the primary care provider can effectively relieve most patients' symptoms and who also ensure that more significant bowel disease is not overlooked (Tab. 3, Ref. 17). PMID- 17125069 TI - Negative affixes in medical English. AB - Many medical terms have negative meaning expressed by means of a negative prefix or suffix. The most frequently used negative prefixes are: a-, dis-, in-, non-, and un-. There is only one negative suffix -less (Ref. 15). PMID- 17125070 TI - Project support of practical training in biophysics. AB - The Department of Biophysics ensures practical training in biophysics and related subjects for students of medical and health study programmes. Demonstrations of medical technology are an important part of this training. Teaching for Faculty of Sciences in biophysical study programmes becomes also very important. Some lectures and demonstrations of technology are involved, but the practical trainig is missing. About 1 mil. CZK for additional laboratory equipment was obtained from the HEIDF project No. 1866/ 2005 "The demonstration and measuring technology for education in medical biophysics and radiological physics" for measuring system DEWETRON for high frequency signal analysis, Fluke Ti30 IR camera, PM 9000B patient monitor, ARSENAL AF 1 fluorescence microscope, and Nikon Coolpix 4500 digital camera with accessories for microphotography. At the present time, further financial resources are being provided by a development project of Ministry of Education "Inter-university co-operation in biomedical technology and engineering using top technologies" in total amount of almost 5 mil CZK, whereas over 2 mil CZK from this project are reserved for student laboratory equipment. The main goal of this project is to ensure the participation of Medical Faculty in educational co-operation in the biomedical technology and engineering, namely with the Faculty of Electrical Engineering and Communication (FEEC), Brno University of Technology. There will be taught those areas of biophysics which are not covered by FEEC, thus forming a separate subject "General Biophysics". The following instruments will be installed: UV-VIS spectrophotometers, rotation viscometers, tensiometers, microscopes with digital image processing, cooled centrifuge, optical benches, and some smaller instruments for practical measurements. PMID- 17125071 TI - [Advisable management protocol for patients with subarachnoidal hemorrhage resulting from cerebral vascular aneurysmal rupture]. PMID- 17125072 TI - [Preliminary results of treatment for spastic forms of infantile cerebral paralysis by chronic epidural neurostimulation of lumbar enlargement]. AB - Six patients with infantile cerebral paralysis following drug-resistant spastic syndrome were operated on. Four children suffered from lower spastic paraparesis; 2 had spastic tetraparesis. All the children were observed to have leg chiasm and myogenic equinus talipes. Electrodes were implanted under X-ray guidance into the posterior epidural cavity of the spinal cord at the level of Th10-Th12 vertebrae and the MATTRIX system. In the postoperative period, all the children had a steady-state decrease in leg and arm muscle tone during 1-2 daily high-frequency electrostimulation sessions. Within the early week, there was a regression of equinus talipes and leg chiasm. A follow-up indicated a steady-state clinical effect in all the patients. Stimulation myography revealed that the H reflex was suppressed and the H/M ratio decreased to the normal level (60-80%) in all the patients. An average of one daily electrostimulation session was sufficient to maintain muscle tone at the near-normal level. PMID- 17125073 TI - [Electrostimulation of a subthalamic nuclear area in Parkinson's disease]. AB - The paper presents the results of application of chronic electrostimulation of the subthalamic nucleus in 15 patients with the akinetic-rigid form of Parkinson's disease. Standard UPDRS, Schwab and England, Hoehn and Yahr, and PDQ 39 rating scales were used for the clinical evaluation of motor disorders. The patients were examined before and 6 months after surgery with ON-stimulation in OFF and ON-medication. Positive results as better life quality, ameliorated motor disorders, and fewer complication of drug therapy with a two-fold reduction in the daily dose of L-dopa were obtained in the vast majority of patients (n = 14). PMID- 17125074 TI - [Temporary clipping and cerebral blood flow in patients with cerebral aneurysms]. AB - Regional cerebral blood flow (RCBF) was measured with cerebral aneurysms by the H2-inhalation technique during intraoperative occlusion test in 15 patients. Temporary clipping was followed by a decrease of RCBF below the critical values in most patients. Reperfusion at the removal of a clip from the internal carotid and middle cerebral artery induced a significant hyperemia (298 +/- 55 ml/100 g x min). At the same time, there was no increase in the frequency of postoperative complications due to temporary clipping. This suggests that their development is related to the duration of cerebral ischemia and the state of the blood-brain barrier. PMID- 17125075 TI - [Local fibrinolytic technique in surgery of traumatic inctracranial hemorrhages]. AB - The emergence of neuroimaging techniques and new surgical technologies (neuroendocopy, navigation systems) in neurosurgery has substantially changed views of surgery for traumatic intracranial hematomas. The local fibrinolytic technique that has been applied to 40 victims aged 18 to 67 years (mean age 42.1 +/- 2 years) who had 18-to-97-cm3 hematomas is a promising direction of mini invasive surgery for traumatic intracranial hematomas in patients in the compensated and subcompensated state. There were 32 males and 8 females. The procedure of the surgical intervention involves drainage of intracranial hematoma, followed by clot lysis and liquid blood aspiration along the drainage. A good outcome with a complete hematoma removal and clinical symptom regression was observed in 26 patients, a fair result with preservation of moderate neurological symptoms at hospital discharge was noted in 2 patients; 3 victims died. Recurrent bleedings were seen in 4 patients with epidural hematomas. A morphological study revealed the typical features of the morphogenesis of traumatic hematomas and perifocal brain tissue during local fibrinolytic therapy, which suggests that the area of damaging effect of bleeding on the adjacent brain tissue is decreased. Local fibrinolysis in surgery of traumatic intracranial hematomas may be considered to be one of the promising lines of treatment policy along with the existing traditional and current techniques and may be used as the method of choice in surgery of traumatic intracranial hematomas in patients in the compensated state. Removal of epidural hematomas through local fibrinolysis should be limited due to a high risk of recurrent hemorrhage and may be made only in a restricted contingent of patients with severe concomitant injury and concurrent somatic diseases when the risk of combined anesthesia and that of a longer operation are rather high. Moreover, of promise is that subtentorial epidural hematomas may be aspirated without trepanation of the posterior cranial fossa and the surgery may be performed under local anesthesia. PMID- 17125076 TI - [Surgical treatment of multileveled diskogenic compression of the cervical spine and its roots]. AB - The authors studied 54 patients with diskogenic compression of the cervical spine and nervous roots, who had undergone interventions including diskectomy, anterior cervical spondylodesis, and plate fixation. The purpose of the study was to provide rationale for choice of a surgical treatment for multileveled diskogenic compression of the cervical spine. Twenty-seven patients with multileveled lesion underwent diskectomy at 2-3 levels with and without vertebral body resection. Preference was given to the mini-invasive technique involving diskectomy, anterior cervical spondylodesis, by using osteoinductor blocks, and plate fixation. A control group consisted of 27 patients with one-leveled lesion who had undergone conventional surgical treatment. 0.5-3-year follow-ups showed positive results in 84% in the study group and in 89% in the control one. Fusion was successful in 89 and 96% in the study and control groups, respectively. The rate of complications and the level of successful professional and social rehabilitation were similar in both groups. PMID- 17125077 TI - [Primary tumors of the central nervous system in the Republic of Tatarstan: clinical epidemiology and medical care organization]. AB - The paper provides data on the incidence of primary tumors of the central nervous system in the Republic of Tatarstan from 1991 to 2000. It also presents sociohygienic characteristics of patients with primary tumors of the brain and spinal cord. The compliance of the diagnosis and treatment of this pathology with state-of-the-art capacities is analyzed. It is concluded that patients were admitted for specialized treatment in the late phases of disease. Early diagnosis and introduction of current methods affecting a pathological process can improve the results of treatment in neurocancer patients in the republic. PMID- 17125078 TI - [Cavernous hemangioma of the optical canal and superior palpebral fissure]. AB - The paper describes a rare case of cavernous hemangioma spreading to the optical canal and superior palpebral fissure. The distinctive feature of the case is the location of cavernous hemangioma in anterior two thirds of the optical canal and superior palpebral fissure. In this connection, despite small sizes of a neoplasm, it caused a significant decrease in visual function during 10 months. The first sign of the disease within 10 months before surgery was periodic diplopia, which served as an indication for magnetic resonance imaging that revealed a pathological process and a correction decision was taken to follow up the patient due to visual preservation. Worse vision served as an indication for surgery as further follow-up might result in irreversible sequels. Undertaken active policy proved its worth despite its risk. The used respective supraorbital access permitted radical removal of a neoplasm to recover visual functions. PMID- 17125079 TI - [Intracranial plasmocytomas: biology, diagnosis, and treatment]. AB - Intracranial plasmocytomas are a rare abnormality in a neurosurgeon's practice. The plasmocytomas may originate from the skull bones or soft tissue intracranial structures; they may be solitary or occur as a manifestation of multiple myeloma, this type being typical of most intracranial plasmocytomas. Progression of solitary plasmocytoma to multiple myeloma is observed in a number of cases. Preoperative diagnosis involves computed tomography or magnetic resonance imaging; angiography is desirable. The final diagnosis of plasmocytoma is chiefly based on a morphological study. Special immunohistochemical studies yield very promising results; these are likely to be of high prognostic value. Intracranial plasmocytomas require a differential approach and a meticulous examination since the presence or absence of multiple myeloma radically affects prognosis. There are well-defined predictors; however, it is appropriate that craniobasal plasmocytomas show a worse prognosis than plasmocytomas of the skull vault and more commonly progress to multiple myeloma. Plasmocytomas respond to radiotherapy very well. The gold standard of treatment for plasmocytoma is its total removal and adjuvant radiation therapy; however, there is evidence for good results when it is partially removed and undergoes radiotherapy or after radical surgery without subsequent radiation. The role of chemotherapy has not been defined today. PMID- 17125080 TI - Safety unbound. PMID- 17125081 TI - Dealing with a hostile employee. PMID- 17125082 TI - Nothing's lost in this translation. PMID- 17125083 TI - Cold case files. PMID- 17125084 TI - Vents' role in improved protection. PMID- 17125085 TI - Blue-ribbon panel zeroes in on culture change. PMID- 17125086 TI - Peabody Energy: changed from the top down. PMID- 17125087 TI - Coping with meth lab hazards. PMID- 17125088 TI - Selecting respirators with confidence. PMID- 17125089 TI - What you don't know. PMID- 17125090 TI - Confined spaces and gas detection. PMID- 17125091 TI - Safety considerations for live electrical measurements. AB - Test instruments available today, when combined with safe work practices, offer troubleshooters many enhancements that make the measurement environment and the measurements themselves much easier and safer than ever before. PMID- 17125094 TI - Sifting through the kids: should children be screened for CKD? PMID- 17125095 TI - KDIGO takes clinical practice guidelines to a global level. PMID- 17125096 TI - IFKF takes global view on care for kidney patients. PMID- 17125097 TI - Katrina aftermath: has the renal community made a full recovery? PMID- 17125098 TI - A descriptive report of errors and adverse events in chronic hemodialysis units. AB - With the combination of technical equipment, medication administration, and caregiver-delivered treatment, opportunities for adverse events and medical errors exist in hemodialysis units. There are no studies describing the type and frequency of medical errors and adverse events in hemodialysis units. This study examines standard adverse events and medical errors reported on routine quality assurance forms by the clinical directors of four hemodialysis units between January 2004 and June 2005. The units varied in size (45 -108 patients), average number of hemodialysis treatments provided (524 -1,333/month), and staffing ratios (1:3 - 1:9.5). In total, 88 errors occurred in 64,541 dialysis treatments (1 event every 733 treatments). Infiltration of the hemodialysis access (n = 31 ) and clotting of the dialysis circuit (n = 19) were also fairly common while dialysis equipment problems occurred relatively rarely (30 occurrences in 64,541 treatments, or 1 event every 2,151 treatments). Thirty-five medication errors occurred (1 every 2,15 1 treatments); omission of an ordered medication was the most common (24/35, 69%). Nine patients fell and six of the falls occurred after a dialysis treatment. No patients required hospitalization as a result of the adverse events or errors. Errors and events were more common in the larger units but did not seem to be directly related to unit staffing ratios. Adverse events in hemodialysis units are fairly common and should be included among routine quality improvement issues addressed by dialysis providers and caregivers. More study of this issue is needed. PMID- 17125099 TI - Improving revenue collections: a primer. PMID- 17125100 TI - Abstracts of the Kidney Disease Economics Conference, October 27-29, 2006, Baltimore, Maryland, USA. PMID- 17125103 TI - Antibiotics: for better or for worse, in sickness and in health, 'til death do us part. PMID- 17125104 TI - Preoperative breast MRI for locoregional staging. AB - BACKGROUND: Magnetic resonance imaging of the breast is indicated in multiple settings, one of which is preoperative staging to establish tumor extent, multicentricity, and contralateral disease. METHODOLOGY: Results of conventional imaging, breast MRI, and pathology were analyzed from 334 patients who underwent breast MRI subsequent to a biopsy that confirmed primary breast malignancy. The focus of this retrospective study was imaging-histologic correlation. FINDINGS: Only 1.2% of patients with tumors mapped by MRI required re-operation after lumpectomy. Multicentricity was identified by MRI alone in 7.7% of patients, while 3.6% were found to have occult contralateral cancer. In 11 of the 12 patients with contralateral cancer, the lesion was invasive, with ductal histology in 9 of the 11. The sensitivity of mammography for detecting these contralateral cancers was only 22.2%. CONCLUSION: Preoperative breast MRI is recommended for mapping tumor extent and to discover additional primary lesions in newly diagnosed breast cancer patients. PMID- 17125105 TI - Part II. Introduction to the beta-lactam antibiotics. PMID- 17125106 TI - Does treatment with antibiotics reduce the duration or severity of symptoms of acute otitis media in children as compared to treatment with analgesics alone? PMID- 17125107 TI - The pinnacle of editorial communications. PMID- 17125108 TI - How to cure U.S. health care. A voucher plan that covers everyone--and helps businesses too. PMID- 17125109 TI - Is Israel ready for disease management? AB - Approximately 60% of all worldwide deaths are caused by chronic disease resulting from modifiable health behaviors. In the United States, structured programs tailored to identify and modify health behaviors of patients with chronic illness have grown into a robust industry called disease management. DM is premised upon the basic assumption that health services utilization and morbidity can be reduced for those with chronic illness by augmenting traditional episodic medical care services and support between physician visits. Given that Israel and the U.S. have similar demographics in their chronically ill populations, it would make intuitive sense for Israel to replicate efforts made in the U.S. to incorporate DM strategies. This paper provides a conceptual framework of how DM could be integrated within the current organizational structure of the Israeli healthcare system, which is uniquely conducive to the implementation of DM on a population-wide basis. While ultimately the decision to invest in DM lies with stakeholders at various institutional levels in Israel, this paper is intended to provide direction and support for that decision-making process. PMID- 17125110 TI - Post-transfusion purpura: a challenging diagnosis. AB - BACKGROUND: Post-transfusion purpura is a rare syndrome characterized by severe thrombocytopenia and bleeding caused by alloimunization to human platelet specific antigens following a blood component transfusion. The suggested incidence is 1:50,000-100,000 transfusions, most often occurring in multiparous women. The diagnosis is not easy because these patients, who are often critically ill or post-surgery, have alternative explanations for thrombocytopenia such as infection, drugs, etc. OBJECTIVES: To describe patients with initially misdiagnosed PTP and to emphasize the diagnostic pitfalls of this disorder. PATIENTS AND RESULTS: During a period of 11 years we diagnosed six patients with PTP, four women and two men. The incidence of PTP was approximately 1:24,000 blood components transfused. We present the detailed clinical course of three of the six patients in whom the diagnosis was particularly challenging. The patients were initially misdiagnosed as having heparin-induced thrombocytopenia, systemic lupus erythematosus complicated by autoimmune thrombocytopenia, and disseminated intravascular coagulation. A history of recent blood transfusion raised the suspicion of PTP and the diagnosis was confirmed by appropriate laboratory workup. CONCLUSIONS: PTP seems to be more frequent than previously described. The diagnosis should be considered in the evaluation of life-threatening thrombocytopenia in both men and women with a recent history of blood transfusion. PMID- 17125111 TI - Transabdominal transanal resection of distal rectal cancer after high dose preoperative radiotherapy: a Chinese experience in preserving sphincter function. AB - BACKGROUND: The combination of high dose preoperative radiotherapy and transanal abdominal transanal radical proctosigmoidectomy and colo-anal anastomosis as a sphincter-preserving method has never been performed in mainland China. OBJECTIVES: To assess the feasibility and efficacy of high dose preoperative radiotherapy and TATA as a sphincter-preserving method in Jiangsu, an economically well-developed region of China with a population of 70 million people. METHODS: From September 1994 to September 2000, 25 consecutive patients with pathologically confirmed distal rectal adenocarcinoma were treated preoperatively with a total dose of 45-46 Gy at 1.8-2.0 Gy per fraction during 5 weeks. Sphincter-preserving surgery by TATA was performed 4-6 weeks after radiotherapy. RESULTS: Acute toxicity of preoperative radiotherapy was tolerable. Eight percent of the patients presented pathologic complete tumor response after preoperative radiotherapy. All patients underwent TATA as scheduled. During a median follow-up of 70 months, the 5 year survival rate was 88%. The 5 year survival rate for those tumors down-staged to pathological TO or to pT1 was 100%. CONCLUSIONS: High dose preoperative radiotherapy and TATA as a sphincter preserving method was feasible and efficient in Chinese patients with distal rectal cancer. In this study, the subset of patients with a good response to radiotherapy had a better clinical outcome. PMID- 17125112 TI - Renal effects of low dose aspirin in elderly patients. AB - BACKGROUND: Aspirin is commonly used by elderly patients. In previous studies we found transient changes in renal function induced by low doses of aspirin. OBJECTIVES: To investigate the mechanisms of these effects. METHODS: The study group included 106 long-term care stable geriatric inpatients. Diet and drugs were kept stable. The study lasted 5 weeks; during the first 2 weeks 100 mg aspirin was administered once a day. Clinical and laboratory follow-up was performed at baseline and weekly for the next 3 weeks. The glomerular filtration rate was estimated by creatinine clearance measured in 24 hour urine and serum creatinine, and by the Cockcroft-Gault formula (C-G) equation. Uric acid clearance was determined from serum concentrations and 24 hour excretion of uric acid. Patients with serum creatinine > 1.5 mg/dl were not included. RESULTS: After 2 weeks on low dose aspirin, measured creatinine and uric acid clearances decreased significantly compared with the initial values in 70% and 62% of the patients, respectively, with mean decreases of 19% and 17%, respectively (P< 0.001). Blood urea nitrogen increased by 17% while serum creatinine and uric acid concentrations increased by 4% (P < 0.05 for all). The C-G values decreased by 3% (P< 0.05). After withdrawal of aspirin all parameters improved. However, 67% of the patients remained with some impairment in their measured Ccr, compared to baseline. Patients who reacted adversely to low dose aspirin had significantly better pre-study renal function (Ccr), lower hemoglobin and lower levels of serum albumin. CONCLUSIONS: Short-term low dose aspirin affected renal tubular creatinine and uric acid transport in the elderly, which may result in a prolonged or permanent deterioration of the renal function. It is suggested that renal functions be monitored even with the use of low dose aspirin in elderly patients. PMID- 17125113 TI - Learning curve in laparoscopic colorectal surgery: our first 100 patients. AB - BACKGROUND: Recent data confirming the oncologic safety of laparoscopic colectomy for cancer as well as its potential benefits will likely motivate more surgeons to perform laparoscopic colorectal surgery. OBJECTIVES: To assess factors related to the learning curve of laparoscopic colorectal surgery, such as the number of operations performed, the type of procedures, major complications, and oncologic resections. METHODS: We evaluated the data of our first 100 elective laparoscopic colorectal operations performed during a 2 year period and compared the first 50 cases with the following 50. RESULTS: The mean age of the study population was 66 years and 49% were males. Indications included cancer, polyps, diverticular disease, Crohn's disease, and others, in 50%, 23%, 13%, 7% and 7% respectively. Mean operative time was 170 minutes. One patient died (massive pulmonary embolism). Significant surgical complications occurred in 10 patients (10%). Hospital stay averaged 8 days. Comparison of the first 50 procedures with the next 50 revealed a significant decrease in major surgical complications (20% vs. 0%). Mean operative time decreased from 180 to 160 minutes and hospital stay from 8.6 to 7.2 days. There was no difference in conversion rate and mean number of harvested nodes in both groups. Residents performed 8% of the operations in the first 50 cases compared with 20% in the second 50 cases. Right colectomies had shorter operative times and fewer conversions. CONCLUSIONS: There was a significant decrease in major complications after the first 50 laparoscopic colorectal procedures. Adequate oncologic resections may be achieved early in the learning curve. Right colectomies are less difficult to perform and are recommended as initial procedures. PMID- 17125114 TI - The benefits and safety of external counterpulsation in symptomatic heart failure. AB - BACKGROUND: External counterpulsation is a safe and effective method of alleviating angina pectoris, but the mechanism of benefit is not understood. OBJECTIVES: To evaluate the safety and efficacy of external counterpulsation therapy in heart failure patients. METHODS: Fifteen symptomatic heart failure patients (subsequent to optimal medical and device therapy) underwent 35 hourly sessions of ECPT over a 7 week period. Before and after each ECPT session we performed pro-B-type natriuretic peptide and brachial artery function studies, administered a quality of life questionnaire, and assessed exercise tolerance and functional class. RESULTS: Baseline left ventricular ejection fraction was 28.1+/ 5.8%. ECPT was safe and well tolerated and resulted in a reduction in pro-BNP levels (from 2,245+/- 2,149 pcg/ml to 1,558+/-1206 pcg/ml, P= 0.022). Exercise duration (Naughton protocol) improved (from 720+/-389 to 893+/-436 seconds, P= 0.0001), along with functional class (2.63+/-0.6 vs. 1.93+/-0.7, P= 0.023) and quality of life scores (54+/-22 vs. 67+/-23, P= 0.001). Nitroglycerine-mediated brachial vasodilatation increased (11.5+/-7.3% vs. 15.6+/-5.2%, P=0.049), as did brachial flow-mediated dilation (8.35+/-6.0% vs. 11.37+/-4.9%, P= 0.09). CONCLUSIONS: ECPT is safe for symptomatic heart failure patients and is associated with functional and neurohormonal improvement. Larger long-term randomized studies with a control arm are needed to confirm these initial encouraging observations. PMID- 17125115 TI - Terlipressin facilitates transport of septic patients treated with norepinephrine. AB - BACKGROUND: Transport of hemodynamic unstable septic patients for diagnostic or therapeutic interventions outside the intensive care unit is complex but sometimes contributes to increasing the chance of survival. OBJECTIVES: To report our experience with terlipressin treatment for facilitation of transport to distant facilities for diagnostic or therapeutic procedures in septic patients treated with norepinephrine. METHODS: We conducted a retrospective analysis of the records of our ICU, identifying the patients with septic shock who required norepinephrine for hemodynamic support. RESULTS: Terlipressin was given to 30 septic shock patients (15 females and 15 males) who were on high dose norepinephrine (10 microg/min or more) in order to facilitate their transport outside the ICU. The dose of terlipressin ranged from 1 to 4 mg, with a mean of 2.13+/-0.68 mg. The dose of norepinephrine needed to maintain systolic blood pressure above 100 mmHg decreased following terlipressin administration, from 21.9+/-10.4 microg/min (range 5-52 microg/min) to 1.0+/-1.95 (range 0-10) (P < 0.001). No patients required norepinephrine dose adjustment during transport. No serious complications or overshoot in blood pressure values were observed following terlipressin administration. Acrocyanosis occurred only in eight patients receiving more than 1 mg of the drug. The overall mortality rate was 50%. CONCLUSIONS: Our data suggest that terlipressin is effective in septic shock. Because it is long-acting and necessitates less titration it might be indicated for patient transportation. PMID- 17125116 TI - Splenic embolus: 13 cases from a single medical department. AB - BACKGROUND: Despite the spleen having a very rich blood supply, there is a paucity of reports of splenic emboli. OBJECTIVES: To investigate the incidence of splenic emboli treated in a single general internal medicine department over the last 3 years. METHODS: We examined the records of a 35 bed internal medicine department in a hospital in the center of Israel. RESULTS: Over a period of 3 years 13 patients admitted to one internal medicine department developed acute abdominal pain and areas of hypoperfusion in the spleen on contrast computed tomography imaging. The patients were treated with anticoagulants, their course was benign and there were no long-term sequelae. CONCLUSIONS: Embolus to the spleen is not rare in an internal medicine department. Diagnosis can be easily made by contrast CT scanning, and treatment with anticoagulants results in a good prognosis. PMID- 17125117 TI - Prevention of early-onset neonatal group B streptococcal infection: is universal screening by culture universally applicable? AB - BACKGROUND: Previous assessments of maternal group B Streptococcus carrier rates in women delivering at Shaare Zedek Medical Center ranged between 3.5 and 11% with neonatal sepsis rates of 0.2-0.9/1000 live births. Because of low colonization and disease rates, routine prenatal cultures of GBS were not recommended and intrapartum prophylaxis was mainly based on maternal risk factors. OBJECTIVES: To determine whether this policy is still applicable. METHODS: We performed prospective sampling and follow-up of women admitted for labor and delivery between February 2002 and July 2002. Vaginal and rectal cultures were obtained before the first pelvic examination. GBS isolation was performed using selective broth medium and identified by latex agglutination and serotyping. Demographic data were collected by means of a standardized questionnaire. Data on the newborns were collected throughout 2002. RESULTS: Of the 629 sampled women, 86 had a positive culture and a carrier rate of 13.7%. A borderline significantly higher carriage rate was observed among mothers of North American origin (21% vs. 13.1%, P= 0.048), and a higher attack rate in their infants (3.8/1000 compared with 0.5/1000 live births in our general maternal population, P= 0.002). Eight newborns had early-onset neonatal GBS sepsis (a rate of 0.8/1000 live births), but none of them benefited from intrapartum antibiotic prophylaxis. CONCLUSIONS: An increased neonatal disease rate was observed in a population with a higher colonization rate than previously seen. In view of the higher carrier rates, we now recommend routine prenatal screening for GBS in our perinatal population. PMID- 17125119 TI - Is disease management "ready" for Israel? PMID- 17125118 TI - Erythropoietin in clinical practice: current use, effect on survival, and future directions. AB - Recombinant human erythropoietin has become an essential part of the management of anemic patients with end-stage renal disease. It is also used to treat the anemia associated with cancer and other diseases, and it improves quality of life. In recent years, studies in animals and humans have focused on the use of rHuEPO for other indications. It has been found to play a role in both cardioprotection and neuroprotection. It has effects on the immune system, and can cause regression in hematologic diseases such as multiple myeloma. It may also improve the response of solid tumors to chemotherapy and radiation therapy. On the other hand, concerns have been raised following two studies of patients with solid tumors in whom those treated with rHuEPO had diminished survival. Criticism of the design of these studies makes it clear that large, well designed, randomized trials must be performed to determine the role of rHuEPO in the treatment of cancer, and more generally to clarify the full clinical benefits of the drug, while minimizing the harm. PMID- 17125120 TI - Post-transfusion purpura--when and why? PMID- 17125121 TI - Rectal cancer: progress over the past two decades. PMID- 17125122 TI - Usefulness of 64 multislice spiral computed tomography for preoperative assessment of severe coarctation of aorta in an adult. PMID- 17125123 TI - Intestinal leishmaniasis in a patient with AIDS. PMID- 17125124 TI - Dull abdominal pain caused by non-functional metastatic neuroendocrine tumor. PMID- 17125125 TI - Giant cholesteatoma of the temporal bone. PMID- 17125127 TI - Dipyrone-induced oligohydramnios and ductus arteriosus restriction. PMID- 17125126 TI - 18F-fluorodeoxyglucose-avid thyroid incidentalomas in patients with lymphoma. PMID- 17125128 TI - Cessation of microemboli in the middle cerebral artery after a single dose of aspirin in a young patient with emboliogenic lacunar syndrome of carotid origin. PMID- 17125135 TI - Fast multidimensional NMR by polarization sharing. AB - The speed of multidimensional NMR spectroscopy can be significantly increased by drastically shortening the customary relaxation delay between scans. The consequent loss of longitudinal magnetization can be retrieved if 'new' polarization is transferred from nearby spins. For correlation spectroscopy involving heteronuclei (X=13C or 15N), protons not directly bound to X can repeatedly transfer polarization to the directly bound protons through Hartmann Hahn mixing. An order of magnitude increase in speed has been observed for the 600 MHz two-dimensional HMQC spectra of amikacin and strychnine using this technique, and it also reduces the noisy F1 ridges that degrade many heteronuclear correlation spectra recorded with short recovery times. PMID- 17125130 TI - The validation process of incorporating simulation-based accreditation into the anesthesiology Israeli national board exams. AB - BACKGROUND: The Israeli Board of Anesthesiology Examination Committee added a simulation-based Objective Structured Clinical Evaluation component to the board examination process. This addition was made in order to evaluate medical competence and considers certain domains that contribute to professionalism. This unique and new process needed to be validated. OBJECTIVES: To validate and evaluate the reliability and realism of incorporating simulation-based OSCE into the Israeli Board Examination in Anesthesia. METHODS: Validation was performed before the exam regarding Content Validity using the modified Delphi technique by members of the Task Force of the Israeli Board Examination Committee in Anesthesiology. RESULTS: The examination has been administered six times in the past 3 years to a total of 145 examinees. The pass rate ranged from 62% (trauma) to 91% (regional anesthesia). The mean inter-rater correlations for the total score (all items), for the Critical checklist items score, and for the Global (General) rating were 0.89, 0.86 and 0.76, respectively. The inter-correlations between the five OSCE stations scores were significant (P< 0.01) only between Trauma & Ventilation for the Total score (r = 0.32, n=63), and between Resuscitation & Regional and OR-crisis for the Global score (r = 0.42 and 0.27, n=64 and 104, respectively). The correlation between the OSCE examination score and the success rate at each of the eight different clinical domains of the oral board examination did not reach statistical significance. Most participants (70 90%) found the difficulty level of the examination stations reasonable to very easy. All major errors, which were identified in the initial two exam periods, disappeared later in the next two exam periods. CONCLUSIONS: The exam has gradually progressed from being an optional part of the oral board examination to a prerequisite component of this test. Other anesthesiology programs or medical professions can adopt the model described here. PMID- 17125129 TI - Levofloxacin-induced interstitial nephritis and vasculitis in an elderly woman. PMID- 17125133 TI - Design and implementation of a web-based, database-driven histology atlas: technology at work. AB - At Vanderbilt University, the "Human Cell and Tissue Biology" course is a required lecture and laboratory course with 2 full-time instructors and 106 students. To address demands placed on faculty for individual attention, an interactive Web-based histology atlas was developed and implemented in January 2005. This atlas was specifically designed to complement the existing laboratory manual and to transform the manual into an interactive educational tool whereby students could view high-resolution images of histological specimens online. By utilizing a computer scripting language, interactive highlighting of histological structures was accomplished through the implementation of a simple mouse-rollover function. This computer-aided instruction software allows students to preview histological structures of interest prior to entering the laboratory, to have additional faculty-directed contact hours during laboratory, and to review material efficiently. The conversion of the originally developed static application into a database-driven tool streamlined the development and modification of the atlas while facilitating the creation of advanced features. Six weeks after launching this interactive atlas, Vanderbilt medical students logged 1,200 hr of use. Through the cooperative efforts of faculty and students, the interactive atlas evolved to meet the educational demands of medical students owing to the development and implementation of a database structure. The functionality and educational value of the interactive atlas in facilitating self learning was ultimately measured by positive student feedback and use. PMID- 17125131 TI - Influence of antibiotic cefotaxime on somatic embryogenesis and plant regeneration in indica rice. AB - An antibiotic, cefotaxime (Omnatax) has been found to promote somatic embryogenesis and subsequent plant regeneration in vitro in indica-type basmati rice cultures. Response was highly genotype specific. The number, mass and morphology of the calli formed on the scutellar tissues were dependent on the growth medium (with or without cefotaxime). The embryogenic nature of nodular calli was confirmed through histological analysis and their plant regeneration ability. The calli of variety Pusa basmati 1 grown on medium supplemented with cefotaxime (100 mg/L) exhibited up to 70.5% plant regeneration as compared to control (51.51%). Plants regenerated from emryogenic calli were phenotypically normal and identical to seed-derived plants and exhibited normal fertility. A limited humidity and an optimal aeration of the culture tubes further enhanced the frequency of somatic embryogenesis and plant regeneration. PMID- 17125140 TI - Leptin: nourishment for the immune system. AB - Leptin, a protein produced by adipocytes, exerts several functions, including modulation of the immune response. A report in this issue of the European Journal of Immunology describes for the first time the effect of either leptin receptor deficiency or blockade on murine dendritic cell (DC) maturation, survival and function. The study describes how leptin receptor deficiency/blockade delays DC maturation and promotes apoptosis, shifts the balance between pro- and anti inflammatory cytokine production, and reduces the ability of DC to stimulate CD4(+) lymphocytes. These exciting novel data add an important piece of evidence to the picture of the role of leptin in immunity and inflammation and generate the possibility that many of the effects of leptin on T lymphocytes might be mediated through DC. PMID- 17125141 TI - Deficiency of tenascin C attenuates allergen-induced bronchial asthma in the mouse. AB - Tenascin C (TN-C) is an extracellular matrix glycoprotein whose expression is increased in several inflammatory diseases of the lung, including bronchial asthma. However, the exact function of TN-C in the pathogenesis of lung inflammation remains unclear. In the present study, we compared the degree of bronchial asthma in wild-type and TN-C-deficient (-/-) BALB/c mice. Bronchial asthma was induced by sensitization and challenge with ovalbumin. Littermates treated with saline were used as controls. Cytokines in bronchoalveolar lavage fluid and plasma were measured by enzyme immunoassays. The number of eosinophils in the lung was significantly increased in wild-type mice compared with TN-C knockout mice. Airway hyperreactivity, NF-kappaB activation and concentrations of monocyte chemoattractant protein-1, IL-5, IL-13, metalloproteinase-9 and immunoglobulin-E in the bronchoalveolar lavage fluid were significantly decreased in ovalbumin-sensitized/challenged TN-C-knockout mice compared with their wild type counterparts. In vitro experiments disclosed that TN-C significantly stimulates the secretion of IL-5, IL-13, IFN-gamma and immunoglobulin-E from spleen lymphocytes. These observations suggest that TN-C is involved in the pathogenesis of bronchial asthma. PMID- 17125142 TI - Role of MHC class II expressing CD4+ T cells in proteolipid protein(91-110) induced EAE in HLA-DR3 transgenic mice. AB - MHC class II molecules play a central role in the control of adaptive immune responses through selection of the CD4(+) T cell repertoire in the thymus and antigen presentation in the periphery. Inherited susceptibility to autoimmune disorders such as multiple sclerosis, rheumatoid arthritis and IDDM are associated with particular MHC class II alleles. Advent of HLA transgenic mice has helped us in deciphering the role of particular HLA DR and DQ class II molecules in human autoimmune diseases. In mice, the expression of class II is restricted to professional antigen-presenting cells (APC). However, in humans, class II is also expressed on T cells, unlike murine T cells. We have developed new humanized HLA class II transgenic mice expressing class II molecules not only on APC but also on a subset of CD4(+) T cells. The expression of class II on CD4(+) T cells is inducible, and class II(+) CD4(+) T cells can present antigen in the absence of APC. Further, using EAE, a well-established animal model of MS, we tested the functional significance of these class II(+) CD4(+) T cells. DR3.AEo transgenic mice were susceptible to proteolipid protein(91-110)-induced EAE and showed CNS pathology accompanied by widespread inflammation and demyelination seen in human MS patients, suggesting a role for class II(+) CD4(+) T cells in the pathogenesis. PMID- 17125143 TI - Involvement of leptin signaling in the survival and maturation of bone marrow derived dendritic cells. AB - Previous studies demonstrated that lymphocyte development is impaired in leptin receptor (Ob-R)-deficient db/db mice. However, it remains unclear whether or not leptin signaling plays a physiological role in dendritic cell (DC) development and function. In this study, we first detected Ob-R expression in murine DC. Using db/db mice at a pre-diabetic stage, we demonstrate that the total number of DC generated from bone marrow (BM) cultures is significantly lower than in WT controls. Similarly, selective blockade of leptin with a soluble mouse Ob-R chimera (Ob-R:Fc) inhibited DC generation in wild-type BM cultures. The reduced DC yield in db/db BM culture was attributed to significantly increased apoptosis, which was associated with dysregulated expression of Bcl-2 family genes. Moreover, db/db DC displayed markedly reduced expression of co-stimulatory molecules and a Th2-type cytokine profile, with a poor capacity to stimulate allogeneic T cell proliferation. Consistent with their impaired DC phenotype and function, db/db DC showed significantly down-regulated activities of the PI3K/Akt pathway as well as STAT-3 and IkappaB-alpha. In conclusion, our findings demonstrate the involvement of leptin signaling in DC survival and maturation. PMID- 17125144 TI - Functional and structural requirements for the internalization of distinct BCR ligand complexes. AB - Antigen (Ag) binding to the BCR rapidly initiates two important events: a phosphorylation cascade that results in the production of secondary signaling intermediaries and the internalization of Ag-BCR complexes. Previous studies using anti-BCR antibodies (Ab) have suggested that BCR signaling is an essential requirement for BCR endocytosis and have further implicated lipid rafts as essential platforms for both BCR functions. However, published data from our laboratory indicate that lipid rafts and consequently raft-mediated signaling are dispensable for BCR-mediated internalization of Ag-specific BCR. Therefore, we investigated the relationship between BCR signaling and endocytosis by defining the role of early kinase signaling in the BCR-mediated internalization of a model Ag (haptenated protein). The results demonstrate that Src kinases and Syk mediated BCR signaling are not essential for BCR-mediated Ag internalization. Moreover, by comparing Ag and Ab, it was determined that while both localize to clathrin-coated pits, the internalization of Ab-BCR complexes is more susceptible to inhibition of signaling and highly sensitive to disruption of lipid rafts and the actin cytoskeleton compared to Ag-BCR complexes. Thus, these results demonstrate that the nature of the ligand ultimately determines the functional requirements and relative contribution of lipid rafts and other membrane structures to the internalization of BCR-ligand complexes. PMID- 17125145 TI - Bcl6 controls granzyme B expression in effector CD8+ T cells. AB - Bcl6, a sequence-specific transcriptional repressor, is important for generation and maintenance of memory CD8(+) T cells. Although memory CD8(+) T cells are generated from effector CD8(+) T cells, a role for Bcl6 in effector CD8(+) T cells is largely unknown. We show here that Bcl6 expression was transiently induced in activated CD8(+) T cells and continuously up-regulated in effector CD8(+) T cells. The amount of granzyme B mRNA among effector molecules produced by effector CD8(+) T cells inversely correlated with the amount of Bcl6 mRNA in CD8(+) T cells. Overexpression of Bcl6 in CD8(+) T cells resulted in lower killing activity at their effector phase, supporting the reduction of granzyme B expression in effector CD8(+) T cells by Bcl6. We identified a putative Bcl6 binding DNA sequence in the promoter region of the granzyme B gene. Binding of Bcl6 to the Bcl6-binding sequence was detected in naive CD8(+) T cells but not in activated CD8(+) T cells by chromatin immunoprecipitation assay. Furthermore, the Bcl6-binding sequence was required for Bcl6 to repress the luciferase reporter gene expression controlled by the granzyme B promoter. Thus, the granzyme B gene is a molecular target of Bcl6 in effector CD8(+) T cells. PMID- 17125146 TI - Parameters governing exhaustion of rare T cell-independent neutralizing IgM producing B cells after LCMV infection. AB - The late appearance of neutralizing antibodies (nAb) against lymphocytic choriomeningitis virus (LCMV) has been attributed to various factors including immunopathology, low frequency of high-affinity specific B cells and competition by nonspecific polyclonal B cell activation. To investigate the activation of LCMV-nAb-producing B cells early following infection, we performed adoptive transfers of LCMV-specific B cells into WT recipients. By modulating parameters such as viral load, number of specific B cells and presence of T cell help, we found that a high antigen-to-B cell ratio led to normal IgM responses. IgG and memory response however, were impaired as most nAb-producing B cells rapidly terminally differentiated into short-lived IgM plasma cells. Lowering the antigen to-B cell ratio, or increasing the level of T cell help, could rescue the class switched antibody response. Upon infection, a low frequency of LCMV-nAb-producing B cells, as observed in WT mice, results in a high antigen-to-B cell ratio and is likely to lead to terminal differentiation - and elimination - of these rare B cells. PMID- 17125149 TI - Disruption of mating in codling moth (Lepidoptera: Tortricidae) by chlorantranilipole, an anthranilic diamide insecticide. AB - The influence of the anthranilic diamide insecticide chlorantranilipole (DPX E2Y45; Rynaxypyr against the adult stage of codling moth, Cydia pomonella L., was examined. Insecticide residues in plastic cups sprayed with 56 mg AI L(-1) (equivalent to the recommended field rate) had a minimal effect on adult survival or fecundity, but significantly fewer female moths were mated in treated than in untreated cups. Similar results were found in field studies with adults placed in screened cages on treated apple foliage for up to 3 weeks. Mating was disrupted when either sex was topically dosed with technical material (0.01-0.1 microg). Males were more sensitive to the disruptive effects of chlorantranilipole than females. A 2 h exposure significantly reduced male activity (walking and wing fanning), but not female calling. A 25 h exposure significantly reduced the activity of both sexes. The proportion of males landing near a sex pheromone source in a flight tunnel was significantly reduced following exposures of 2-4 h and completely eliminated following a 17 h exposure. Male moths partially recovered from either topical or residual exposure to chlorantranilipole, but the extent of their recovery was dose and time dependent. Male response to sex pheromone recovered within 2 h following a 4 h exposure, but following a 17 h exposure only a partial recovery occurred after 48 h. Mating by moth pairs exposed to chlorantranilipole residues for 24 h did not significantly increase during recovery periods of 24-96 h, except in cups treated with 10% of the standard concentration. The proportion of field-collected female codling moths caught in pear ester-baited traps that were mated was significantly lower in apple orchards treated season-long with chlorantranilipole compared with females caught in either untreated orchards or those treated with sex pheromones to cause mating disruption. PMID- 17125150 TI - Survey of the year 2005 commercial optical biosensor literature. AB - We identified 1113 articles (103 reviews, 1010 primary research articles) published in 2005 that describe experiments performed using commercially available optical biosensors. While this number of publications is impressive, we find that the quality of the biosensor work in these articles is often pretty poor. It is a little disappointing that there appears to be only a small set of researchers who know how to properly perform, analyze, and present biosensor data. To help focus the field, we spotlight work published by 10 research groups that exemplify the quality of data one should expect to see from a biosensor experiment. Also, in an effort to raise awareness of the common problems in the biosensor field, we provide side-by-side examples of good and bad data sets from the 2005 literature. PMID- 17125151 TI - Selection of colour of sticky trap for monitoring adult bean thrips, Caliothrips fasciatus (Thysanoptera: Thripidae). AB - Adult bean thrips, Caliothrips fasciatus (Pergande), overwintering inside the navel of navel oranges shipped from California to Australia, are an actionable pest for the importing country, i.e. infested lots are fumigated with methyl bromide. Strict quarantine regulations regarding C. fasciatus prompted studies on the best colour sticky trap that might be used to monitor for bean thrips populations in the vicinity of California citrus groves prior to harvesting fruit for export. Preliminary experiments identified the most attractive trap of each of four colours (blue, green, white, yellow) commonly used to sample adult Thysanoptera. Three trials of a field study were conducted, comparing C. fasciatus capture on the best card of each colour using asparagus ferns naturally infested with high levels of this pest. Based on significantly higher catch on green sticky cards, this colour trap is recommended for potential use in California's bean thrips mitigation plan designed to reduce thrips levels on citrus exported to Australia. PMID- 17125153 TI - Demonstrating formation of potentially persistent transformation products from the herbicides bromoxynil and ioxynil using liquid chromatography-tandem mass spectrometry (LC-MS/MS). AB - It is shown that potentially persistent transformation products can be formed from the herbicides bromoxynil (3,5-dibromo-4-hydroxybenzonitrile) and ioxynil (3,5-diiodo-4-hydroxybenzonitrile), and possible leaching to groundwater is discussed. A similar process to the formation of BAM (2,6-dichlorobenzamide) from the herbicide dichlobenil (2,6-dichlorobenzonitrile) can be anticipated as bromoxynil and ioxynil are analogues of dichlobenil and they are degraded by the enzymes nitrilase, nitrile hydratase and amidase. A biodegradation study using cultured Variovorax sp. DSM 11402, a species commonly found in soil, demonstrated that ioxynil and bromoxynil were fully transformed into their corresponding amides in 2-5 days. These amides were not further degraded within 18 days, and formation of other degradation products was not observed. These results are in agreement with biodegradation experiments with dichlobenil. In soil, dichlobenil is transformed into its only observed degradation product BAM, which is persistent and mobile, and has been found in 19% of 5000 samples of Danish groundwater. Variovorax sp. is known to degrade the non-halogenated analogue benzamide, suggesting that degradation of the three amides may be hindered by the halogenated substituents (meta-Br; meta-I; ortho-Cl). This hypothesis is supported by QSAR modelling of fundamental properties. Using a new optimised liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, the sorption and desorption properties of bromoxynil and ioxynil were characterised in sandy topsoil at four concentration levels. The estimated sorption coefficient K(d) was 1.4 L kg(-1) for bromoxynil and 5.4 L kg(-1) for ioxynil, indicating weak to moderate sorption to topsoil. Desorption of the herbicides showed that they were strongly and irreversible bound to the soil (K(des) > K(d)). The amount of herbicide desorbed depended on the initial concentration level. At low levels, K(des) values were higher, indicating stronger binding than at higher levels. The isocratic LC-MS/MS method developed for simultaneous detection of bromoxynil, ioxynil and their main degradation products is described. Using negative electrospray ionisation (ESI-), the detection limits were 0.4-1.0 microg L(-1), with relative standard deviations of 4-10% (n = 10) using direct injection without clean-up steps. The standard curves showed linearity in the range 5-100 microg L(-1) with r(2) > 0.992. PMID- 17125152 TI - Desorption/ionization on porous silicon mass spectrometry (DIOS) of model cationized fatty acids. AB - Desorption/ionization on porous silicon (DIOS) is a very useful technique in the case of small molecular weight compounds, compared to the matrix-assisted laser desorption ionization (MALDI). This is because MALDI generates matrix-related ions that overlap with the mass range of interest. The aim of our work was to investigate the suitability of the DIOS technique in the case of fatty acids in negative ion mode. The analysis of the chosen fatty acid models, nonadecanoic acid (C(19)H(38)O(2)) and heneicosanoic acid (C(21)H(42)O(2)), gave rise to the observation of the deprotonated monomeric species and selective cationized multimeric species. This cation selectivity was further elucidated by complementary studies based on the addition of various metals such as Ag(I), Zn(II), Fe(II), and also Cu(II). Specific behavior, depending upon the introduced metal, was highlighted by different redox reaction processes and also metastable decompositions (in PSD mode). PMID- 17125154 TI - Esterase-mediated bifenthrin resistance in a multiresistant strain of the two spotted spider mite, Tetranychus urticae. AB - A field-collected multiresistant strain of Tetranychus urticae Koch exhibiting high resistance to bifenthrin was investigated in comparison with a susceptible laboratory strain. The esterase inhibitor S,S,S-tributyl-phosphorotrithioate (DEF) was able strongly to synergise bifenthrin toxicity in the resistant strain. Optimal conditions for determining esterase activities in T. urticae were determined, and a higher esterase activity towards several artificial substrates was found in this resistant strain, which had a preference for hydrolysing 4 nitrophenyl butyrate. Bifenthrin was able to bind the active centres of T. urticae esterases in vitro, as was determined after competition experiments by a Dixon plot, revealing a higher affinity of bifenthrin in the resistant strain. Bifenthrin-hydrolysing activity in the resistant and susceptible strains was examined in vitro and quantified with gas chromatography. A 7.2-fold higher metabolising rate was found in the resistant strain. PMID- 17125155 TI - A simple purification protocol for the detection of peptide hormones in the hemolymph of individual insects by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - The endocrine system of insects is largely based on peptide hormones. Nevertheless, an unequivocal chemical demonstration of the occurence in the hemolymph (the 'insect blood') is still lacking for most if not all insect peptide hormones, although this is the only way to prove their hormonal status. Focusing on peptides released during ecdysis behavior of the tobacco hornworm Manduca sexta, we developed a purification protocol based on ultrafiltration and a single reversed-phase high-performance liquid chromatography (RP-HPLC) step that for the first time allowed the mass spectrometric and chemical identification of a peptide hormone in the hemolymph of single specimens. Since this method is simple, relatively cheap and fast, it should be useful for routine endocrinological analyses and for monitoring peptide release during different physiological conditions and behaviors in insects. PMID- 17125156 TI - Application of phenylboronic acid modified hydrogel affinity chips for high throughput mass spectrometric analysis of glycated proteins. AB - The complexity of the human plasma proteome is greatly increased by post translational modifications. Besides physiological modifications, pathological conditions such as diabetes are responsible for adding to this complexity by producing advanced glycation endproducts (AGEs). When searching for specific biomarkers it is a prerequisite to reduce this complexity prior to analysis. To do this, agarose hydrogel was used to create a high-capacity affinity layer on the modified aluminum surface of MALDI (matrix-assisted laser desorption/ionization) targets. 3-Aminophenylboronic acid was immobilized via cyanogen bromide activation as a ligand for affinity sorption of glycated proteins, followed by their direct detection by MALDI. High protein capacity of prepared MALDI chips, efficient separation and low non-specific protein binding were demonstrated. The results show that phenylboronic acid modified hydrogels are very suitable for creating affinity surfaces for the high-throughput analysis of AGEs. PMID- 17125158 TI - Rationale for a model of human systems integration: the need of a theoretical framework. AB - Human systems integration (HSI) involves augmented human design with the objectives of augmenting human capabilities and improving human performance using behavioral technologies. The fundamental matter of human systems integration and augmented human design is the organization and the nature of interactions that couple physiological systems, humans- and engineered systems, artifacts. By this definition, augmented human consists of interactive artefacts linked to physiological systems. This paper focuses on the rationale of a HSI model based on specific experiments (comparison of dynamical sensorimotor integration and motor performances in real and virtual environments) that confirm the hypothesis of functional interaction in the framework of Chauvet's mathematical theory of integrative physiology (MTIP). Epistemological constraints for HSI and the role of MTIP are briefly discussed in this context. PMID- 17125157 TI - Are sparse-coding simple cell receptive field models physiologically plausible? AB - Olshausen and Field (1996) developed a simple cell receptive field model for natural scene processing in V1, based on unsupervised learning and non-orthogonal basis function optimization of an overcomplete representation of visual space. The model was originally tested with an ensemble of whitened natural scenes, simulating pre-cortical filtering in the retinal ganglia and lateral geniculate nucleus, and the basis functions qualitatively resembled the orientation-specific responses of V1 simple cells in the spatial domain. In this study, the quantitative tuning responses of the basis functions in the spectral domain are estimated using a Gaussian model, to determine their goodness-of-fit to the known bandwidths of simple cells in primate V1. Five simulation experiments which examined key features of the model are reported: changing the size of the basis functions; using a complete versus over-complete representation; changing the sparseness factor; using a variable learning rate; and mapping the basis functions with a whitening spatial function. The key finding of this study is that across all image themes, basis function sizes, number of basis functions, sparseness factors and learning rates, the spatial-frequency tuning did not closely resemble that of primate area 17 -- the model results more closely resembled the unclassified cat neurones of area 19 with a single exception, and not area 17 as predicted. PMID- 17125159 TI - Quantum mechanics and brain uncertainty. AB - This paper argues that molecular governing structures (such as receptors, gating molecules, or ionic channels) which operate pervasively in the brain, often with small number particle systems (as, for example, at the surfaces of membranes, synaptic clefts, or macromolecules), may plausibly be vehicles for the transmutation of quantum mechanical fluctuations to normal-level neural signaling. PMID- 17125160 TI - A new paradigm for theory in integrative biology: the principle of auto associative stabilization: biochemical networks and the selection of neuronal groups. AB - This paper discusses rationale for a theory in biology: what exactly is a theory in biology? Is it of a mathematical nature? How to conceive an integrative theory and why? Replies to these questions are offered for subsequent discussions as concerns the mathematical theory of integrative physiology (MTIP) proposed by the author. It is shown that such a theory is a theoretical framework built on a representation in terms of hierarchical functional interactions and a specific formalism, the S-Propagator, to traverse the levels of organization. As for all natural theories, the MTIP is based on a general principle specific to biology, the principle of auto-associative stabilization (PAAS). In this framework, two models are revisited for a novel interpretation: the first addresses the dynamics of biochemical networks, the second addresses the selection of groups of neurons (TSGN) as suggested by Edelman. PMID- 17125161 TI - On the existence of physiological age based on functional hierarchy: a formal definition related to time irreversibility. AB - The present approach of aging and time irreversibility is a consequence of the theory of functional organization that I have developed and presented over recent years (see e.g., Ref. 11). It is based on the effect of physically small and numerous perturbations known as fluctuations, of structural units on the dynamics of the biological system during its adult life. Being a highly regulated biological system, a simple realistic hypothesis, the time-optimum regulation between the levels of organization, leads to the existence of an internal age for the biological system, and time-irreversibility associated with aging. Thus, although specific genes are controlling aging, time-irreversibility of the system may be shown to be due to the degradation of physiological functions. In other words, I suggest that for a biological system, the nature of time is specific and is an expression of the highly regulated integration. An internal physiological age reflects the irreversible course of a living organism towards death because of the irreversible course of physiological functions towards dysfunction, due to the irreversible changes in the regulatory processes. Following the works of Prigogine and his colleagues in physics, and more generally in the field of non integrable dynamical systems (theorem of Poincare-Misra), I have stated this problem in terms of the relationship between the macroscopic irreversibility of the functional organization and the basic mechanisms of regulation at the lowest "microscopic" level, i.e., the molecular, lowest level of organization. The neuron-neuron elementary functional interaction is proposed as an illustration of the method to define aging in the nervous system. PMID- 17125162 TI - On the integration of physiological mechanisms in the nervous tissue using the MTIP: synaptic plasticity depending on neurons-astrocytes-capillaries interactions. AB - The objective in this work is twofold: (i) to illustrate the use of the Mathematical Theory of Integrative Physiology (MTIP) [13], that is a general theory and practical method for the systematic and progressive mathematical integration of physiological mechanisms; (ii) to study a complex neurobiological system taken as an example, i.e., the synaptic plasticity depending on brain activity, on astrocytic and neuronal metabolism, and on brain hemodynamics. The functional organization of the nervous tissue is presented in the framework of the MTIP, the ultimate objective being the study of learning and memory by coupling the three networks of neurons, astrocytes and capillaries. Specifically in this paper, we study the influence of the variation of capillaries arterial oxygen on the induction of LTP/LTD by coupling validated mathematical models of AMPA, NMDA, VDCC channels, calcium current in the dendritic spine, vesicular glutamate dynamics in the presynaptic bouton derived from glycolysis and neuronal glucose, mitochondrial respiration, Ca/Na pumps, glycolysis, and calcium dynamics in the astrocytes, hemodynamics of the capillaries. The integration of all these models is discussed by claiming the advantages of using a common framework and a specific dedicated computing system, PhysioMatica. PMID- 17125163 TI - Johann Gasteiger--Germany's pioneer in chemoinformatics. PMID- 17125164 TI - Analysis of data fusion methods in virtual screening: theoretical model. AB - This paper presents a theoretical model of how data fusion can be used to combine the results of multiple similarity searches of chemical databases. The model is based on frequency distributions of similarity values that are fused using a multiple integration over regions defined by the particular fusion rule that is being applied. For pairwise fusion, the resulting double integrals are straightforward to evaluate for simple model distributions. Similarity values for recovered-active and recovered-nonactive frequency distributions are independently modeled using a constant background, linearly biased terms, and a first-order correlated term. The model shows that two standard fusion rules can give performance enhancements in some cases but that the results of fusion are dependent on many factors that, taken together, can lead to seemingly inconsistent levels of enhancement. PMID- 17125165 TI - Analysis of data fusion methods in virtual screening: similarity and group fusion. AB - In a recent companion paper we have related the operation of simple data fusion rules used in virtual screening to a multiple integral formalism. In this paper we extend these ideas to the analysis of data fusion methods applied to real data. We examine several cases of similarity fusion using different coefficients and different representations and consider the reasons for positive or negative results in terms of the similarity distributions. Results are obtained using the SUM-, MAX- MIN-, and CombMNZ-fusion rules. We also develop a customized fusion rule, which provides an estimate of the optimal possible result for fusing multiple searches of a specific database; this shows that similarity fusion can, in principle, achieve retrieval enhancements even if this is not achieved in practice with current fusion rules. The methods are extended to analyze the comparatively successful results of group fusion with multiple actives, and we provide a rationale for the observed superiority of the MAX-rule over the SUM rule in this context. PMID- 17125166 TI - NIPALSTREE: a new hierarchical clustering approach for large compound libraries and its application to virtual screening. AB - A hierarchical clustering algorithm--NIPALSTREE--was developed that is able to analyze large data sets in high-dimensional space. The result can be displayed as a dendrogram. At each tree level the algorithm projects a data set via principle component analysis onto one dimension. The data set is sorted according to this one dimension and split at the median position. To avoid distortion of clusters at the median position, the algorithm identifies a potentially more suited split point left or right of the median. The procedure is recursively applied on the resulting subsets until the maximal distance between cluster members exceeds a user-defined threshold. The approach was validated in a retrospective screening study for angiotensin converting enzyme (ACE) inhibitors. The resulting clusters were assessed for their purity and enrichment in actives belonging to this ligand class. Enrichment was observed in individual branches of the dendrogram. In further retrospective virtual screening studies employing the MDL Drug Data Report (MDDR), COBRA, and the SPECS catalog, NIPALSTREE was compared with the hierarchical k-means clustering approach. Results show that both algorithms can be used in the context of virtual screening. Intersecting the result lists obtained with both algorithms improved enrichment factors while losing only few chemotypes. PMID- 17125167 TI - Chemoinformatics: past, present, and future. AB - The history of chemoinformatics is reviewed in a decade-by-decade manner from the 1940s to the present. The focus is placed on four traditional research areas: chemical database systems, computer-assisted structure elucidation systems, computer-assisted synthesis design systems, and 3D structure builders. Considering the fact that computer technology has been one of the major driving forces of the development of chemoinformatics, each section will start from a brief description of the new advances in computer technology of each decade. The summary and future prospects are given in the last section. PMID- 17125168 TI - Prediction of pKa values for aliphatic carboxylic acids and alcohols with empirical atomic charge descriptors. AB - Two quantitative pKa prediction models for aliphatic carboxylic acids and for alcohols were developed by multiple linear-regression (MLR) analysis with empirical atomic descriptors. The acid and alcohol molecules were described by a set of five and four atomic descriptors, respectively. For the pKa model of 1122 aliphatic carboxylic acids, the squared correlation coefficient is 0.813 with a standard error of prediction of 0.423; for the pKa model of 288 alcohols, the squared correlation coefficient is 0.817 with a standard error of prediction of 0.755, respectively. The good predictive abilities of the models obtained were indicated by both cross-validation and by external validation. An atomic descriptor was developed to model the inductive effect of the neighboring atoms for a central atom in a molecule. The ability of the descriptor to measure the inductive effect of substituent groups was demonstrated by a good correlation of this descriptor with Taft sigma* constants in aliphatic carboxylic acids. It provides a new approach to estimate Taft sigma* constants directly from molecular structures. An algorithm using Kohonen neural networks for splitting a data set into a training set and a test set is also presented. PMID- 17125169 TI - Basic overview of chemoinformatics. AB - There is no particular point in time that determines when chemoinformatics was founded or established. It slowly evolved from several, often quite humble beginnings. Scientists in various fields of chemistry struggled with the development of computer methods which allowed them to manage the enormous amount of chemical information and to find relationships between the structure and properties of a compound. During the 1960s some early developments appeared that led to a flurry of activities in the 1970s. This review provides a general overview of basic methods in the specific fields of chemoinformatics, from encoding chemical compounds, storing and searching data in databases, to generating and analyzing these data. In addition, the chief interconnecting points of chemoinformatics applications are highlighted including the contributions of Johann Gasteiger to this field. PMID- 17125170 TI - Physicochemical stereodescriptors of atomic chiral centers. AB - Physicochemical atomic stereodescriptors (PAS) were implemented that represent the chirality of an atomic chiral center on the basis of empirical physicochemical properties of the ligands. The ligands are ranked according to a specific property, and the chiral center takes an S/R-like descriptor relative to that property. The procedure is performed for a series of properties, yielding a chirality profile. Application of the PAS descriptors to the prediction of enantioselectivity in chemical reactions, from the molecular structures, is illustrated here. The relationship between the molecular structures, represented by the PAS descriptors, and the enantioselectivity was learned by neural networks, decision trees, or random forests. In a first application, a data set was employed with chiral amino alcohols that enantioselectively catalyze the addition of diethylzinc to benzaldehyde. Prediction of the major enantiomer obtained in the reaction, from the molecular structure of the catalyst, was achieved with accuracy up to 90%. The second application investigated the enantiopreference of Pseudomonas cepacia lipase (PCL) toward primary alcohols. The learned models could make correct predictions about the preferred enantiomer, from the molecular structure of the substrate, in up to 93% of the cases. These included substrates with and without O-atoms bonded to the chiral center. The properties automatically selected to build the models can give indications on the relevant factors guiding the observed chemical behavior. PMID- 17125171 TI - Algorithm for exhaustive and nonredundant organic stereoisomer generation. AB - Generation of organic stereoisomers with R/S, Z/E, and/or M/P configurations that may contain heteroatoms, multiple bonds, and any kind of cycle (isolated, spiro, condensed, and nested) is described. Inputs for processing are molecular structures in a N_tuple format resident on an automatic (canonical) or manual (non canonical) generated file which are processed by doing internal molecular graph construction, a weighted bipartite tree construction for all atoms and bonds to detect stereocenters, and symmetrical atom groups (SAG) with some specific SAG parameters that constitute a novel way for redundancy elimination of meso structures. Finally, determination of ligand CIP priorities allows for writing the output N_tuples with stereoisomer description. Several examples showing application of this methology to a wide number of structures are also presented. PMID- 17125172 TI - Modeling of Gibbs energy of formation of organic compounds by linear and nonlinear methods. AB - Two quantitative models for the prediction of the Gibbs energy of formation (DeltaGf degrees ) of 177 organic compounds were developed. These molecules contain elements such as H, C, N, O, F, S, Cl, and Br, with the molecular weight in the range of 16.04-202.25. The molecules were represented by six selected 2D structure descriptors. At first, the complex relationship between DeltaGf degrees and the six selected input descriptors was depicted by a two-dimensional Kohonen's self-organizing neural network (KohNN) map; on the basis of the KohNN map, the whole data set was split into a training set consisting of 130 compounds and a test set (or a validation set and a test set) including 47 compounds. Then, DeltaGf degrees was predicted using a multilinear regression (MLR) analysis and a back-propagation (BPG) neural network. For 177 organic compounds, root-mean square deviations of 17.8 and 15.4 kcal mol-1 were achieved by MLR and the BPG neural network, respectively. PMID- 17125173 TI - MIMUMBA revisited: torsion angle rules for conformer generation derived from X ray structures. AB - A method has been developed which automatically generates SMARTS patterns for four-atomic torsional fragments, searches experimental structures in the Cambridge Crystallographic Database, and obtains rules for preferred torsion angles in drug-size molecules. These rules can be used for exhaustive conformational analysis using the popular conformer generator OMEGA. This approach results in an overall improvement of quality and coverage of conformational space when comparing conformer ensembles generated by this method with results obtained by using the default OMEGA setup. In particular, the percentage of structures with at least one conformation closer than 0.5 A to the X-ray structure improves from 84% to 92% in a test set of 11 027 experimental structures from the CSD. Moreover, the average RMS distance of the closest conformation to the X-ray structure improves from 0.30 to 0.22 A. PMID- 17125174 TI - Modeling robust QSAR. AB - Quantitative Structure Activity Relationship (QSAR) is a term describing a variety of approaches that are of substantial interest for chemistry. This method can be defined as indirect molecular design by the iterative sampling of the chemical compounds space to optimize a certain property and thus indirectly design the molecular structure having this property. However, modeling the interactions of chemical molecules in biological systems provides highly noisy data, which make predictions a roulette risk. In this paper we briefly review the origins for this noise, particularly in multidimensional QSAR. This was classified as the data, superimposition, molecular similarity, conformational, and molecular recognition noise. We also indicated possible robust answers that can improve modeling and predictive ability of QSAR, especially the self organizing mapping of molecular objects, in particular, the molecular surfaces, a method that was brought into chemistry by Gasteiger and Zupan. PMID- 17125175 TI - Applications of self-organizing neural networks in virtual screening and diversity selection. AB - Artificial neural networks provide a powerful technique for the analysis and modeling of nonlinear relationships between molecular structures and pharmacological activity. Many network types, including Kohonen and counterpropagation, also provide an intuitive method for the visual assessment of correspondence between the input and output data. This work shows how a combination of neural networks and radial distribution function molecular descriptors can be applied in various areas of industrial pharmaceutical research. These applications include the prediction of biological activity, the selection of screening candidates (cherry picking), and the extraction of representative subsets from large compound collections such as combinatorial libraries. The methods described have also been implemented as an easy-to-use Web tool, allowing chemists to perform interactive neural network experiments on the Novartis intranet. PMID- 17125176 TI - Impact of conformational flexibility on three-dimensional similarity searching using correlation vectors. AB - Many three-dimensional (3D) virtual screening concepts, like automated docking or pharmacophore searching, rely on the calculation of a "bioactive" or "receptor relevant" conformation of a molecule to assess its biological activity. We investigated the dependence of the presence of conformations near the "bioactive" conformation on three-dimensional similarity searching with pharmacophore-based correlation vectors (CATS3D approach). Cocrystal structures of 11 target classes served as queries for virtual screening of a database of annotated ligands. Different numbers of conformations were calculated. Single 3D structures were obtained using the 3D structure generator CORINA and conformational ensembles by the conformation generation program ROTATE. This approach was able to reproduce conformations for high resolution cocrystal structures. For virtual screening we found that using only the CORINA-generated single conformation already resulted in a significant enrichment of isofunctional molecules having the same biological property profile. This observation was also made for ligand classes with many rotatable bonds. Although more similar conformations were considered to be more similar in the CATS3D description, the impact of using multiple conformations on the enrichment of actives was not as high as expected. CATS3D provides an alignment-free three-dimensional virtual screening approach that is less dependent on the presence of conformations which are close to the "bioactive" conformation of a molecule compared to methods that rely on an explicit three dimensional alignment of molecules. PMID- 17125177 TI - Query generation to search for inhibitors of enzymatic reactions. AB - A method for the generation of intermediates of enzyme-catalyzed reactions is presented. These intermediates can be used as three-dimensional structural queries for searching for inhibitors of enzymatic reactions. The intermediates can be considered as being structurally quite close to transition-state analogues. For this application, a database containing detailed chemical information on metabolic reactions is used. The likely three-dimensional structure of the intermediates of enzyme-catalyzed reactions can be generated from the information in the database. For three reactions catalyzed by the enzymes AMP deaminase (EC code 3.5.4.6), triose phosphate isomerase (EC code 5.3.1.1), and arginase II (EC code 3.5.3.1), we show how a 3D model of these intermediates can be superimposed onto known inhibitors of these enzymes by a program that uses a genetic algorithm. For this, we test different methods for the superimposition using information on the enzymatic binding site, using information on physicochemical properties calculated from the molecular structure, or without having any information in the superimposition process. We show that these inhibitors are most similar to the corresponding intermediates regarding the 3D structure. PMID- 17125178 TI - The impact of tautomer forms on pharmacophore-based virtual screening. AB - In the field of in silico screening, many applications do not automatically consider possible tautomeric states of molecules. However, the detection of new compound candidates might rely on correct structural description, which is important for the perfect fit toward the biologically relevant interactions. In this paper, we present a new exhaustive tautomer enumeration approach implemented by means of the CACTVS software package. The approach contains a set of 21 predefined SMIRKS-based transforms and a powerful transformation engine that is capable of generating most tautomers described comprehensively in the literature or found in databases in the field of medicinal chemistry. User-defined tautomer rules applied to specific structural databases or scientific issues can be implemented easily and used instead of the predefined rules. In addition, we describe the impact of tautomer-enriched databases on pharmacophore screening approaches for human matrix metalloproteinase 8 as an example of a protein-based pharmacophore screening scenario and for human cyclin-dependent kinases as an example of a ligand-based pharmacophore screening approach. In both test cases, as a preprocessing step, we have used our new tautomer enumerator tool for the tautomer enrichment of the screening data sets and have used it as a postprocessing step to remove tautomeric duplicates from the results. We could demonstrate that the tautomer-enriched screening data sets show significant advantages compared to their non-enhanced counterparts. The discrimination between hits and nonhits was significantly better in the case of tautomer enriched databases. Moreover, it has been proved that tautomer-enhanced databases will lead to a higher number of potential hits. PMID- 17125179 TI - Design and development of chemical ontologies for reaction representation. AB - This paper describes the development of chemical ontologies applied to the representation of organic chemical reactions. The ontologies are built using the methodology known as methontology. The hierarchically structured set of terms describing the subdomains, namely, organic reactions, organic compounds, and reagents, are constructed into individual ontologies. The ontologies consist of about 200 concepts and around 125 individuals. A set of binary relations is defined in order to integrate the ontologies with applications. The ontologies are implemented as an XML application with a set of vocabulary describing the domain knowledge. This paper also features an easy-to-use chemical ontological support system (COSS) intended to represent organic chemical reactions automatically. As a model application, the automatic representation of aliphatic nucleophilic substitution reactions is demonstrated using COSS. The paper also describes a keyword-based search system whose functionality is backed with COSS. PMID- 17125180 TI - Chemoinformatics-based classification of prohibited substances employed for doping in sport. AB - Representative molecules from 10 classes of prohibited substances were taken from the World Anti-Doping Agency (WADA) list, augmented by molecules from corresponding activity classes found in the MDDR database. Together with some explicitly allowed compounds, these formed a set of 5245 molecules. Five types of fingerprints were calculated for these substances. The random forest classification method was used to predict membership of each prohibited class on the basis of each type of fingerprint, using 5-fold cross-validation. We also used a k-nearest neighbors (kNN) approach, which worked well for the smallest values of k. The most successful classifiers are based on Unity 2D fingerprints and give very similar Matthews correlation coefficients of 0.836 (kNN) and 0.829 (random forest). The kNN classifiers tend to give a higher recall of positives at the expense of lower precision. A naive Bayesian classifier, however, lies much further toward the extreme of high recall and low precision. Our results suggest that it will be possible to produce a reliable and quantitative assignment of membership or otherwise of each class of prohibited substances. This should aid the fight against the use of bioactive novel compounds as doping agents, while also protecting athletes against unjust disqualification. PMID- 17125181 TI - Learning from the data: mining of large high-throughput screening databases. AB - High-throughput screening (HTS) campaigns in pharmaceutical companies have accumulated a large amount of data for several million compounds over a couple of hundred assays. Despite the general awareness that rich information is hidden inside the vast amount of data, little has been reported for a systematic data mining method that can reliably extract relevant knowledge of interest for chemists and biologists. We developed a data mining approach based on an algorithm called ontology-based pattern identification (OPI) and applied it to our in-house HTS database. We identified nearly 1500 scaffold families with statistically significant structure-HTS activity profile relationships. Among them, dozens of scaffolds were characterized as leading to artifactual results stemming from the screening technology employed, such as assay format and/or readout. Four types of compound scaffolds can be characterized based on this data mining effort: tumor cytotoxic, general toxic, potential reporter gene assay artifact, and target family specific. The OPI-based data mining approach can reliably identify compounds that are not only structurally similar but also share statistically significant biological activity profiles. Statistical tests such as Kruskal-Wallis test and analysis of variance (ANOVA) can then be applied to the discovered scaffolds for effective assignment of relevant biological information. The scaffolds identified by our HTS data mining efforts are an invaluable resource for designing SAR-robust diversity libraries, generating in silico biological annotations of compounds on a scaffold basis, and providing novel target family specific scaffolds for focused compound library design. PMID- 17125182 TI - SemanticEye: a semantic web application to rationalize and enhance chemical electronic publishing. AB - SemanticEye, an ontology with associated tools, improves the classification and open accessibility of chemical information in electronic publishing. In a manner analogous to digital music management, RDF metadata encoded as Adobe XMP can be extracted from a variety of document formats, such as PDF, and managed in an RDF repository called Sesame. Users upload electronic documents containing XMP to a central server by "dropping" them into WebDAV folders. The documents can then be navigated in a Web browser via their metadata, and multiple documents containing identical metadata can then be aggregated. SemanticEye does not actually store any documents. By including unique identifiers within the XMP, such as the DOI, associated documents can be retrieved from the Web with the help of resolving agents. The power of this metadata driven approach is illustrated by including, within the XMP, InChI identifiers for molecular structures and finding relationships between articles based on their InChIs. SemanticEye will become increasingly more comprehensive as usage becomes more widespread. Furthermore, following the Semantic Web architecture enables the reuse of open software tools, provides a "semantically intuitive" alternative to search engines, and fosters a greater sense of trust in Web-based scientific information. PMID- 17125183 TI - Melting point prediction employing k-nearest neighbor algorithms and genetic parameter optimization. AB - We have applied the k-nearest neighbor (kNN) modeling technique to the prediction of melting points. A data set of 4119 diverse organic molecules (data set 1) and an additional set of 277 drugs (data set 2) were used to compare performance in different regions of chemical space, and we investigated the influence of the number of nearest neighbors using different types of molecular descriptors. To compute the prediction on the basis of the melting temperatures of the nearest neighbors, we used four different methods (arithmetic and geometric average, inverse distance weighting, and exponential weighting), of which the exponential weighting scheme yielded the best results. We assessed our model via a 25-fold Monte Carlo cross-validation (with approximately 30% of the total data as a test set) and optimized it using a genetic algorithm. Predictions for drugs based on drugs (separate training and test sets each taken from data set 2) were found to be considerably better [root-mean-squared error (RMSE)=46.3 degrees C, r2=0.30] than those based on nondrugs (prediction of data set 2 based on the training set from data set 1, RMSE=50.3 degrees C, r2=0.20). The optimized model yields an average RMSE as low as 46.2 degrees C (r2=0.49) for data set 1, and an average RMSE of 42.2 degrees C (r2=0.42) for data set 2. It is shown that the kNN method inherently introduces a systematic error in melting point prediction. Much of the remaining error can be attributed to the lack of information about interactions in the liquid state, which are not well-captured by molecular descriptors. PMID- 17125184 TI - Novel 2D fingerprints for ligand-based virtual screening. AB - This paper describes the development of a set of new 2D fingerprints for the purposes of virtual screening in a pharmaceutical environment. The new fingerprints are based on established ones: the changes in their design included the introduction of overlapping pharmacophore feature types, feature counts for pharmacophore and structural fingerprints, as well as changes in the resolution in property description for property fingerprints. The effects of each of these changes on virtual screening performance were monitored using two types of training sets, emulating different stages in the drug discovery process. The results demonstrate that these changes all lead to an improvement in virtual screening performance. PMID- 17125185 TI - SMIREP: predicting chemical activity from SMILES. AB - Most approaches to structure-activity-relationship (SAR) prediction proceed in two steps. In the first step, a typically large set of fingerprints, or fragments of interest, is constructed (either by hand or by some recent data mining techniques). In the second step, machine learning techniques are applied to obtain a predictive model. The result is often not only a highly accurate but also hard to interpret model. In this paper, we demonstrate the capabilities of a novel SAR algorithm, SMIREP, which tightly integrates the fragment and model generation steps and which yields simple models in the form of a small set of IF THEN rules. These rules contain SMILES fragments, which are easy to understand to the computational chemist. SMIREP combines ideas from the well-known IREP rule learner with a novel fragmentation algorithm for SMILES strings. SMIREP has been evaluated on three problems: the prediction of binding activities for the estrogen receptor (Environmental Protection Agency's (EPA's) Distributed Structure-Searchable Toxicity (DSSTox) National Center for Toxicological Research estrogen receptor (NCTRER) Database), the prediction of mutagenicity using the carcinogenic potency database (CPDB), and the prediction of biodegradability on a subset of the Environmental Fate Database (EFDB). In these applications, SMIREP has the advantage of producing easily interpretable rules while having predictive accuracies that are comparable to those of alternative state-of-the-art techniques. PMID- 17125186 TI - "Bayes affinity fingerprints" improve retrieval rates in virtual screening and define orthogonal bioactivity space: when are multitarget drugs a feasible concept? AB - Conventional similarity searching of molecules compares single (or multiple) active query structures to each other in a relative framework, by means of a structural descriptor and a similarity measure. While this often works well, depending on the target, we show here that retrieval rates can be improved considerably by incorporating an external framework describing ligand bioactivity space for comparisons ("Bayes affinity fingerprints"). Structures are described by Bayes scores for a ligand panel comprising about 1000 activity classes extracted from the WOMBAT database. The comparison of structures is performed via the Pearson correlation coefficient of activity classes, that is, the order in which two structures are similar to the panel activity classes. Compound retrieval on a recently published data set could be improved by as much as 24% relative (9% absolute). Knowledge about the shape of the "bioactive chemical universe" is thus beneficial to identifying similar bioactivities. Principal component analysis was employed to further analyze activity space with the objective to define orthogonal ligand bioactive chemical space, leading to nine major (roughly orthogonal) activity axes. Employing only those nine activity classes, retrieval rates are still comparable to original Bayes affinity fingerprints; thus, the concept of orthogonal bioactive ligand chemical space was validated as being an information-rich but low-dimensional representation of bioactivity space. Correlations between activity classes are a major determinant to gauge whether the desired multitarget activity of drugs is (on the basis of current knowledge) a feasible concept because it measures the extent to which activities can be optimized independently, or only by strongly influencing one another. PMID- 17125187 TI - Fuzzy tricentric pharmacophore fingerprints. 1. Topological fuzzy pharmacophore triplets and adapted molecular similarity scoring schemes. AB - This paper introduces a novel molecular description--topological (2D) fuzzy pharmacophore triplets, 2D-FPT--using the number of interposed bonds as the measure of separation between the atoms representing pharmacophore types (hydrophobic, aromatic, hydrogen-bond donor and acceptor, cation, and anion). 2D FPT features three key improvements with respect to the state-of-the-art pharmacophore fingerprints: (1) The first key novelty is fuzzy mapping of molecular triplets onto the basis set of pharmacophore triplets: unlike in the binary scheme where an atom triplet is set to highlight the bit of a single, best matching basis triplet, the herein-defined fuzzy approach allows for gradual mapping of each atom triplet onto several related basis triplets, thus minimizing binary classification artifacts. (2) The second innovation is proteolytic equilibrium dependence, by explicitly considering all of the conjugated acids and bases (microspecies). 2D-FPTs are concentration-weighted (as predicted at pH=7.4) averages of microspecies fingerprints. Therefore, small structural modifications, not affecting the overall pharmacophore pattern (in the sense of classical rule based assignment), but nevertheless triggering a pKa shift, will have a major impact on 2D-FPT. Pairs of almost identical compounds with significantly differing activities ("activity cliffs" in classical descriptor spaces) were in many cases predictable by 2D-FPT. (3) The third innovation is a new similarity scoring formula, acknowledging that the simultaneous absence of a triplet in two molecules is a less-constraining indicator of similarity than its simultaneous presence. It displays excellent neighborhood behavior, outperforming 2D or 3D two point pharmacophore descriptors or chemical fingerprints. The 2D-FPT calculator was developed using the chemoinformatics toolkit of ChemAxon (www.chemaxon.com). PMID- 17125188 TI - Substructure-based support vector machine classifiers for prediction of adverse effects in diverse classes of drugs. AB - Unforeseen adverse effects exhibited by drugs contribute heavily to late-phase failure and even withdrawal of marketed drugs. Torsade de pointes (TdP) is one such important adverse effect, which causes cardiac arrhythmia and, in some cases, sudden death, making it crucial for potential drugs to be screened for torsadogenicity. The need to tap the power of computational approaches for the prediction of adverse effects such as TdP is increasingly becoming evident. The availability of screening data including those in organized databases greatly facilitates exploration of newer computational approaches. In this paper, we report the development of a prediction method based on a support machine vector algorithm. The method uses a combination of descriptors, encoding both the type of toxicophore as well as the position of the toxicophore in the drug molecule, thus considering both the pharmacophore and the three-dimensional shape information of the molecule. For delineating toxicophores, a novel pattern recognition method that utilizes substructures within a molecule has been developed. The results obtained using the hybrid approach have been compared with those available in the literature for the same data set. An improvement in prediction accuracy is clearly seen, with the accuracy reaching up to 97% in predicting compounds that can cause TdP and 90% for predicting compounds that do not cause TdP. The generic nature of the method has been demonstrated with four data sets available for carcinogenicity, where prediction accuracies were significantly higher, with a best receiver operating characteristics (ROC) value of 0.81 as against a best ROC value of 0.7 reported in the literature for the same data set. Thus, the method holds promise for wide applicability in toxicity prediction. PMID- 17125189 TI - New global communication process in thermodynamics: impact on quality of published experimental data. AB - Thermodynamic data are a key resource in the search for new relationships between properties of chemical systems that constitutes the basis of the scientific discovery process. In addition, thermodynamic information is critical for development and improvement of all chemical process technologies. Historically, peer-reviewed journals are the major source of this information obtained by experimental measurement or prediction. Technological advances in measurement science have propelled enormous growth in the scale of published thermodynamic data (almost doubling every 10 years). This expansion has created new challenges in data validation at all stages of the data delivery process. Despite the peer review process, problems in data validation have led, in many instances, to publication of data that are grossly erroneous and, at times, inconsistent with the fundamental laws of nature. This article describes a new global data communication process in thermodynamics and its impact in addressing these challenges as well as in streamlining the delivery of the thermodynamic data from "data producers" to "data users". We believe that the prolific growth of scientific data in numerous and diverse fields outside thermodynamics, together with the demonstrated effectiveness and versatility of the process described in this article, will foster development of such processes in other scientific fields. PMID- 17125190 TI - Adaptive configuring of radial basis function network by hybrid particle swarm algorithm for QSAR studies of organic compounds. AB - The configuring of a radial basis function network (RBFN) consists of selecting the network parameters (centers and widths in RBF units and weights between the hidden and output layers) and network architecture. The issues of suboptimum and overfitting, however, often occur in RBFN configuring. This paper presented a hybrid particle swarm optimization (HPSO) algorithm to simultaneously search the optimal network structure and parameters involved in the RBFN (HPSORBFN) with an ellipsoidal Gaussian function as a basis function. The continuous version of PSO was used for parameter training, while the modified discrete PSO was employed to determine the appropriate network topology. The proposed HPSORBFN algorithm was applied to modeling the inhibitory activities of substituted bis[(acridine-4 carboxamide)propyl]methylamines to murine P388 leukemia cells and the bioactivities of COX-2 inhibitors. The results were compared with those obtained from RBFNs with the parameters optimized by continuous PSO and by conventionally RBFN training the algorithm for a fixed network topology, indicating that the HPSO was competent for RBFN configuring in that it converged quickly toward the optimal solution and avoided overfitting. PMID- 17125191 TI - Emerging chemical patterns: a new methodology for molecular classification and compound selection. AB - A concept termed Emerging Chemical Patterns (ECPs) is introduced as a novel approach to molecular classification. The methodology makes it possible to extract key molecular features from very few known active compounds and classify molecules according to different potency levels. The approach was developed in light of the situation often faced during the early stages of lead optimization efforts: too few active reference molecules are available to build computational models for the prediction of potent compounds. The ECP method generates high resolution signatures of active compounds. Predictive ECP models can be built based on the information provided by sets of only three molecules with potency in the nanomolar and micromolar range. In addition to individual compound predictions, an iterative ECP scheme has been designed. When applied to different sets of active molecules, iterative ECP classification produced compound selection sets with increases in average potency of up to 3 orders of magnitude. PMID- 17125192 TI - Design and evaluation of a novel class-directed 2D fingerprint to search for structurally diverse active compounds. AB - Recent attempts to increase similarity search performance using molecular fingerprints have mostly focused on the evaluation of alternative similarity metrics or scoring schemes, rather than the development of new types of fingerprints. Here, we introduce a novel 2D fingerprint design (property descriptor value range-derived fingerprint or PDR-FP) that involves activity oriented selection of property descriptors and the transformation of descriptor value ranges into a binary format such that each fingerprint bit position represents a specific value interval. The design is tailored toward multiple template similarity searching and permits training on specific activity classes. In search calculations on 15 compound classes of increasing structural diversity, the PDR fingerprint performed better than other state-of-the-art 2D fingerprints. Among the structurally diverse classes were six compound sets with peptide character, which represent a notoriously difficult chemotype for 2D similarity searching. In these cases, PDR-FP produced promising results, whereas other fingerprint methods mostly failed. PDR-FP is specifically designed for search calculations on structurally diverse compounds, and these calculations are not influenced by molecular size effects, which represent a general problem for similarity searching using bit string representations. PMID- 17125193 TI - Role of aromacity, ring strain, and stereochemistry of selected disulfides and their congeners in the oxidative linkage of DNA strands at the major groove. A computational study. AB - Three aromatic disulfides and their two selected congeners were studied in order to see if they could be initiators or catalysts in a process that aims to tie together the two strands of a DNA double helix. Thermodynamic functions were calculated from first principle molecular computations on deoxyribose models and the disulfide congener. The gas reaction of one of the disulfides turned out to be the closest to the thermoneutral process. Solvation study caused a relatively minor energetic modification. The disulfides were subjected to docking studies, and the stabilities of the complexes obtained were further analyzed by molecular dynamics simulations. Only one of the two atropic disulfide enantiomers was the matching partner of the chiral DNA double helix, and therefore this remained in the major groove of the DNA. The other enantiomer dissociated from the major groove as it was incapable of forming a stable complex with the chiral double helix. A mechanistic scheme for the reaction has been investigated. PMID- 17125194 TI - Representation of chemical information in OASIS centralized 3D database for existing chemicals. AB - The present inventory of existing chemicals in regulatory agencies in North America and Europe, encompassing the chemicals of the European Chemicals Bureau (EINECS, with 61 573 discrete chemicals); the Danish EPA (159 448 chemicals); the U.S. EPA (TSCA, 56 882 chemicals; HPVC, 10 546 chemicals) and pesticides' active and inactive ingredients of the U.S. EPA (1379 chemicals); the Organization for Economic Cooperation and Development (HPVC, 4750 chemicals); Environment Canada (DSL, 10851 chemicals); and the Japanese Ministry of Economy, Trade, and Industry (16811), was combined in a centralized 3D database for existing chemicals. The total number of unique chemicals from all of these databases exceeded 185 500. Defined and undefined chemical mixtures and polymers are handled, along with discrete (hydrolyzing and nonhydrolyzing) chemicals. The database manager provides the storage and retrieval of chemical structures with 2D and 3D data, accounting for molecular flexibility by using representative sets of conformers for each chemical. The electronic and geometric structures of all conformers are quantum-chemically optimized and evaluated. Hence, the database contains over 3.7 million 3D records with hundreds of millions of descriptor data items at the levels of structures, conformers, or atoms. The platform contains a highly developed search subsystem--a search is possible on Chemical Abstracts Service numbers; names; 2D and 3D fragment searches; structural, conformational, or atomic properties; affiliation in other chemical databases; structure similarity; logical combinations; saved queries; and search result exports. Models (collections of logically related descriptors) are supported, including information on a model's author, date, bioassay, organs/tissues, conditions, administration, and so forth. Fragments can be interactively constructed using a visual structure editor. A configurable database browser is designed for the inspection and editing of all types of data items. Database statistics are maintained on the number and quality of structures, conformers, and descriptors. Reports can be generated presenting any chosen subset of structures and descriptors into different formats suitable for inclusion into documents. In addition to fixed report formats, there is a powerful report template designer module with a visual report template editor to produce a customized page layout. The database is compatible at the import/export level with SDF, MOL, SMILES, and other known formats. The precalculated centralized 3D database could be useful for quantitative structure-activity relationship developers avoiding the time consuming and cumbersome 3D calculation phase of model development. PMID- 17125195 TI - Prediction of multiple binding modes of the CDK2 inhibitors, anilinopyrazoles, using the automated docking programs GOLD, FlexX, and LigandFit: an evaluation of performance. AB - Anilinopyrazoles as CDK2 inhibitors can adopt multiple binding modes depending on the substituents at the 5-position of the pyrazole ring, based on CDK2/cyclin A crystallographic studies. Three commercially available docking programs, FlexX, GOLD, and LigandFit, were tested with 63 anilinopyrazole analogues in an attempt to reproduce the binding modes observed in the crystal structures. Each docking program gave different ligand conformations depending on the scoring or energy functions used. FlexX/drugscore, GOLD/chemscore, and LigandFit/plp were the best combinations of each docking program in reproducing the ligand conformations observed in the crystal structures. The 63 analogues were divided into two groups, type-A and type-B, depending on the substituent at the 5-position of the pyrazole ring. Although an alternate binding mode, observed in a crystal structure of one type-B compound, could not be reproduced with any of the above docking/scoring combinations, GOLD, with a template constraint based on the crystal structure coordinates, was able to reproduce the pose. As for type-A compounds, all docking conditions yielded similar poses to those observed in crystal structures. When predicting activities by scoring programs, the combination of docking with LigandFit/plp and scoring with LIGSCORE1_CFF gave the best correlation coefficient (r=0.60) between experimental pIC50 values and top ranked rescores of 30 poses of each compound. With regard to type-A compounds, the correlation was 0.69. However, when 11 compounds, whose top-ranked rescored poses did not demonstrate the correct binding modes in reference to the crystal structure, were removed, the correlation rose to 0.75. Consequently, predicting activity on the basis of correct binding modes was found to be reliable. PMID- 17125196 TI - A novel method to simulate the hydrophobic effect and its application to the ranking of host/guest complexes. AB - Solvent entropy change is a major factor in driving the association of hydrophobic species in aqueous solutions. We have developed a novel methodology which simulates the solvation of hydrophobic surfaces by water. A system of virtual solvent particles surrounding the solute governed by arbitrarily determined rules provides a means to estimate the degree of order (Q) imposed by such solvation. Computed changes in Q (dQ) upon complex formation have been found to correlate well with observed binding affinities of host-guest complexes in aqueous solutions. Examples are described which illustrate the ability of dQ calculations to identify the correct ligand pose from a set of decoy complexes as well as provide rank ordering of a set of highly diverse ligand-protein complexes. Comparisons to surface-area-based calculations are discussed. The Q methodology holds great promise in the development of predictive structure-based approaches to drug design, as it provides a relatively simple means to estimate the hydrophobic effect. PMID- 17125197 TI - Retro-leapfrog and related retro map operations. AB - Operations on maps are well-known theoretical tools for transforming a given polyhedral tessellation. Several theoretical investigations of fullerenes, such as their pi-electronic structure and stability, need information on the original map which was transformed into a larger molecular structure. In this respect, retro-operations, particularly those of the most used leapfrog, chamfering, and capra operations, appear particularly useful in searching the associate graphs of fullerenes. A series of analyzed cages proved to be leapfrog transforms of smaller cages. This information was useful in understanding their closed pi electronic structure and related properties including the local aromaticity. An index based on the optimized geometries enabled the evaluation of aromaticity of their various substructures. Pictorial images of the pi-electron distribution as the main Kekule valence structures have been performed by the aid of the JSCHEM software package. PMID- 17125198 TI - 3D QSAR studies on protein tyrosine phosphatase 1B inhibitors: comparison of the quality and predictivity among 3D QSAR models obtained from different conformer based alignments. AB - A set of 65 flexible peptidomimetic competitive inhibitors (52 in the training set and 13 in the test set) of protein tyrosine phosphatase 1B (PTP1B) has been used to compare the quality and predictive power of 3D quantitative structure activity relationship (QSAR) comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) models for the three most commonly used conformer-based alignments, namely, cocrystallized conformer based alignment (CCBA), docked conformer-based alignment (DCBA), and global minima energy conformer-based alignment (GMCBA). These three conformers of 5 [(2S)-2-({(2S)-2-[(tert-butoxycarbonyl)amino]-3-phenylpropanoyl}amino)3-oxo-3 pentylamino)propyl]-2-(carboxymethoxy)benzoic acid (compound number 66) were obtained from the X-ray structure of its cocrystallized complex with PTP1B (PDB ID: 1JF7), its docking studies, and its global minima by simulated annealing. Among the 3D QSAR models developed using the above three alignments, the CCBA provided the optimal predictive CoMFA model for the training set with cross validated r2 (q2)=0.708, non-cross-validated r2=0.902, standard error of estimate (s)=0.165, and F=202.553 and the optimal CoMSIA model with q2=0.440, r2=0.799, s=0.192, and F=117.782. These models also showed the best test set prediction for the 13 compounds with predictive r2 values of 0.706 and 0.683, respectively. Though the QSAR models derived using the other two alignments also produced statistically acceptable models in the order DCBA>GMCBA in terms of the values of q2, r2, and predictive r2, they were inferior to the corresponding models derived using CCBA. Thus, the order of preference for the alignment selection for 3D QSAR model development may be CCBA>DCBA>GMCBA, and the information obtained from the CoMFA and CoMSIA contour maps may be useful in designing specific PTP1B inhibitors. PMID- 17125199 TI - Statistical modeling of a ligand knowledge base. AB - A range of different statistical models has been fitted to experimental data for the Tolman electronic parameter (TEP) based on a large set of calculated descriptors in a prototype ligand knowledge base (LKB) of phosphorus(III) donor ligands. The models have been fitted by ordinary least squares using subsets of descriptors, principal component regression, and partial least squares which use variables derived from the complete set of descriptors, least angle regression, and the least absolute shrinkage and selection operator. None of these methods is robust against outliers, so we also applied a robust estimation procedure to the linear regression model. Criteria for model evaluation and comparison have been discussed, highlighting the importance of resampling methods for assessing the robustness of models and the scope for making predictions in chemically intuitive models. For the ligands covered by this LKB, ordinary least squares models of descriptor subsets provide a good representation of the data, while partial least squares, principal component regression, and least angle regression models are less suitable for our dual aims of prediction and interpretation. A linear regression model with robustly fitted parameters achieves the best model performance over all classes of models fitted to TEP data, and the weightings assigned to ligands during the robust estimation procedure are chemically intuitive. The increased model complexity when compared to the ordinary least squares linear model is justified by the reduced influence of individual ligands on the model parameters and predictions of new ligands. Robust linear regression models therefore represent the best compromise for achieving statistical robustness in simple, chemically meaningful models. PMID- 17125200 TI - Prediction of pH-dependent aqueous solubility of druglike molecules. AB - In the present work, the Henderson-Hasselbalch (HH) equation has been employed for the development of a tool for the prediction of pH-dependent aqueous solubility of drugs and drug candidates. A new prediction method for the intrinsic solubility was developed, based on artificial neural networks that have been trained on a druglike PHYSPROP subset of 4548 compounds. For the prediction of acid/base dissociation coefficients, the commercial tool Marvin has been used, following validation on a data set of 467 molecules from the PHYSPROP database. The best performing network for intrinsic solubility predictions has a cross validated root mean square error (RMSE) of 0.70 log S-units, while the Marvin pKa plug-in has an RMSE of 0.71 pH-units. A data set of 27 drugs with experimentally determined pH-solubility curves was assembled from the literature for the validation of the combined pH-dependent model, giving a mean RMSE of 0.79 log S units. Finally, the combined model has been applied on profiling the solubility space at low pH of five large vendor libraries. PMID- 17125201 TI - An efficient in silico screening method based on the protein-compound affinity matrix and its application to the design of a focused library for cytochrome P450 (CYP) ligands. AB - A new method has been developed to design a focused library based on available active compounds using protein-compound docking simulations. This method was applied to the design of a focused library for cytochrome P450 (CYP) ligands, not only to distinguish CYP ligands from other compounds but also to identify the putative ligands for a particular CYP. Principal component analysis (PCA) was applied to the protein-compound affinity matrix, which was obtained by thorough docking calculations between a large set of protein pockets and chemical compounds. Each compound was depicted as a point in the PCA space. Compounds that were close to the known active compounds were selected as candidate hit compounds. A machine-learning technique optimized the docking scores of the protein-compound affinity matrix to maximize the database enrichment of the known active compounds, providing an optimized focused library. PMID- 17125202 TI - Recognition of LXXLL by ligand binding domain of the Farnesoid X receptor in molecular dynamics simulation. AB - The Farnesoid X receptor (FXR) has recently become a potential therapeutical target. The recruitment of coactivator protein (specified by LXXLL sequence) is the initial step in transcriptional activation of nuclear receptors (NRs). In this paper, the process of recognition of the LXXLL motif by the ligand binding domain (LBD) of FXR is observed in a 25 ns molecular dynamics simulation. The hydrophobic and hydrogen bonding interactions between the LBD and the coactivator are fully analyzed. This observation provides justification for the 'on deck' model proposed by Nettles and Greene. At last, insight to the protein-polypeptide interactions and protein conformational changes are discussed. PMID- 17125203 TI - Structure-based identification of small molecule binding sites using a free energy model. AB - We separately have shown that the maximal druglike affinity of a given binding site on a protein can be calculated on the basis of the binding-site structure alone by using a desolvation-based free energy model along with the notion that druglike ligands fall into certain physiochemical property ranges. Here, we present an approach where we reformulate the calculated druggability affinity as an additive free energy to facilitate the searching of whole protein surfaces for druglike binding sites. The highest-scoring patches in many cases represent known ligand-binding sites for druggable targets, but not for difficult targets. This approach differs from other approaches in that it does not simply identify pockets with the greatest volume but instead identifies pockets that are likely to be amenable to druglike small-molecule binding. Combining the method with a functional residue prediction method called SCA (statistical coupling analysis) results in the prediction of potentially druggable allosteric binding sites on p38alpha kinase. PMID- 17125204 TI - In silico prediction of blood-brain barrier permeation using the calculated molecular cross-sectional area as main parameter. AB - The cross-sectional area, AD, of a compound oriented in an amphiphilic gradient such as the air-water or lipid-water interface has previously been shown to be crucial for membrane partitioning and permeation, respectively. Here, we developed an algorithm that determines the molecular axis of amphiphilicity and the cross-sectional area, ADcalc, perpendicular to this axis. Starting from the conformational ensemble of each molecule, the three-dimensional conformation selected as the membrane-binding conformation was the one with the smallest cross sectional area, ADcalcM, and the strongest amphiphilicity. The calculated, ADcalcM, and the measured, AD, cross-sectional areas correlated linearly (n=55, slope, m=1.04, determination coefficient, r2=0.95). The calculated cross sectional areas, ADcalcM, were then used together with the calculated octanol water distribution coefficients, log D7.4, of the 55 compounds (with a known ability to permeate the blood-brain barrier) to establish a calibration diagram for the prediction of blood-brain barrier permeation. It yielded a limiting cross sectional area (ADcalcM=70 A2) and an optimal range of octanol-water distribution coefficients (-1.40.50). In the initial data set of 56 molecules, 18 were classified as strong binders (on the basis of the above criteria), and all of those were recovered in the top 22 molecular hits from ROCS. Additionally, computationally generated values of log P were shown to provide a reasonable estimate of the fraction unbound in microsomes, providing the compounds were in their basic form at physiological pH. PMID- 17125207 TI - Genetic algorithm-optimized QSPR models for bioavailability, protein binding, and urinary excretion. AB - In this work, a genetic algorithm (GA) was applied to build up a set of QSPR (quantitative structure-property relationship) models for human absolute oral bioavailability, plasma protein binding, and urinary excretion using the counts of molecular fragments as descriptors. For a pharmacokinetic property, the consensus score of a set of models (20 or 30) was found to improve the correlation coefficient and reduce the standard error significantly. Key fragments that may boost or reduce pharmacokinetic properties were also identified. Databases searches were performed for a set of key fragments identified by bioavailability models. The percentage of hit rates of bioavailability-boosting fragments were significantly higher than those of bioavailability-reducing fragments for MDDR (MDL Drug Data Report), a database of drugs and drug leads entered or entering development. On the other hand, the opposite trend was observed for ACD (Available Chemicals Directory), a database of all kinds of available compounds. PMID- 17125208 TI - Pocket v.2: further developments on receptor-based pharmacophore modeling. AB - A deriving pharmacophore model from the three-dimensional structure of a target protein provides helpful information for analyzing protein-ligand interactions and further improvement of ligand binding affinity. A standalone program, Pocket v.2, has been developed based on the original Pocket module in the de novo drug design program LigBuilder. Pocket v.2 is able to derive a pharmacophore model directly from a given protein-ligand complex structure without human intervention. Key features in the pharmacophore model are automatically reduced to a reasonable number. Pocket v.2 has been applied to several case studies, including cyclin dependent kinase 2, HIV-1 protease, estrogen receptor, and 17beta-hydroxysteroid dehydrogenase. It well reproduced previously published pharmacophore models in all of these cases. One notable feature of Pocket v.2 is that it can tolerate minor conformational changes on the protein side upon binding of different ligands to give a consistent pharmacophore model. For different proteins accommodating the same ligand, Pocket v.2 gives similar pharmacophore models, which opens the possibility to classify proteins with their binding features. PMID- 17125209 TI - The importance of local chemical structure for chemical metabolism by human uridine 5'-diphosphate-glucuronosyltransferase. AB - The uridine 5'-diphosphate- (UDP-)glucuronosyltransferase (UGT) family of enzymes catalyzes the conjugation of chemicals containing a suitable nucleophilic atom with glucuronic acid. Despite the importance of glucuronidation as an elimination and detoxification mechanism for drugs, environmental chemicals, and endogenous compounds, the structural features of substrates that confer isoform selectivity are poorly understood. The relationship between the local molecular structure of nucleophilic atoms of chemicals and the ability of UGT isoforms to glucuronidate the nucleophilic atoms was investigated here. The proximity of an aromatic ring to the nucleophilic atom was highly associated with a greater likelihood of glucuronidation by most UGT isoforms. Similarly, most UGT isoforms were found to have a statistically significant preference for oxygen over nitrogen as the nucleophilic atom. The converse was established only for UGT1A4. Naive Bayes models were trained to predict the site of glucuronidation for eight UGT isoforms on the basis of the partial charge and Fukui function of the nucleophilic atom and whether an aromatic ring was attached to the nucleophilic atom. On average, the cross-validated sensitivity and specificity of the models were approximately 75-80%. For all but UGT2B7, the area under the receiver operating characteristics curve of the model was greater than 0.8, indicating strong predictive ability. A chemical diversity analysis of the currently available data indicates bias toward chemicals containing phenolic groups, and it is likely that the availability of chemical data sets with greater diversity will facilitate further insights into the structural features of substrates that confer enzyme selectivity. PMID- 17125210 TI - Synergistic use of compound properties and docking scores in neural network modeling of CYP2D6 binding: predicting affinity and conformational sampling. AB - Cytochrome P450 2D6 (CYP2D6) is used to develop an approach for predicting affinity and relevant binding conformation(s) for highly flexible binding sites. The approach combines the use of docking scores and compound properties as attributes in building a neural network (NN) model. It begins by identifying segments of CYP2D6 that are important for binding specificity, based on structural variability among diverse CYP enzymes. A family of distinct, low energy conformations of CYP2D6 are generated using simulated annealing (SA) and a collection of 82 compounds with known CYP2D6 affinities are docked. Interestingly, docking poses are observed on the backside of the heme as well as in the known active site. Docking scores for the active site binders, along with compound-specific attributes, are used to train a neural network model to properly bin compounds as strong binders, moderate binders, or nonbinders. Attribute selection is used to preselect the most important scores and compound specific attributes for the model. A prediction accuracy of 85+/-6% is achieved. Dominant attributes include docking scores for three of the 20 conformations in the ensemble as well as the compound's formal charge, number of aromatic rings, and AlogP. Although compound properties were highly predictive attributes (12% improvement over baseline) in the NN-based prediction of CYP2D6 binders, their combined use with docking score attributes is synergistic (net increase of 23% above baseline). Beyond prediction of affinity, attribute selection provides a way to identify the most relevant protein conformation(s), in terms of binding competence. In the case of CYP2D6, three out of the ensemble of 20 SA-generated structures are found to be the most predictive for binding. PMID- 17125211 TI - An integrated in silico analysis of drug-binding to human serum albumin. AB - Approaches such as quantitative structure-activity relationships (QSAR) and molecular modeling are integrated with the study of complex networks to understand drug binding to human serum albumin (HSA). A robust QSAR model using the topological substructural molecular descriptors/design (TOPS-MODE) approach has been derived and shows good predictability and interpretability in terms of structural contribution to drug binding to HSA. A perfect agreement exists between the group/fragment contributions found by TOPS-MODE and the specific interactions of drugs with HSA. These results indicate a preponderant contribution of hydrophobic regions of drugs to the specific binding to drug binding sites 1 and 2 in HSA and specific roles of polar groups which anchor drugs to HSA binding sites. The occurrence of fragments contributing to drug binding to HSA can be represented by complex networks. The fragment-to-fragment complex network displays "small-world" and "scale-free" characteristics and in this way is similar to other complex networks including biological, social, and technological networks. A small number of fragments appear very frequently in most drugs. These molecular "empathic" fragments are good candidates for guiding future drug discovery research. PMID- 17125212 TI - Ligand-based approach to in silico pharmacology: nuclear receptor profiling. AB - Bioactive ligands are a valuable and increasingly accessible source of information about protein targets. On the basis of this statement, a list of 25 nuclear receptors was described by a series of bioactive ligands extracted directly from bibliographical sources, stored properly in an annotated chemical library, and mathematically represented using the recently reported SHED molecular descriptors. Analysis of this ligand information allowed for derivation of a threshold of nuclear receptor concern. If the similarity of one molecule to any of the molecules annotated to one particular nuclear receptor is below that threshold, the molecule receives an alert on the probability of having affinity below 10 microM for that nuclear receptor. On this basis, a linkage map was constructed that reveals the interaction network of nuclear receptors from the perspective of their active ligands. This ligand-based approach to nuclear receptor profiling was subsequently applied to four external chemical libraries of 10,000 molecules targeted to proteases, kinases, ion channels, and G protein coupled receptors. The percentage of each library that returned an alert on at least one nuclear receptor was reasonably low and varied between 4.4 and 9.7%. In addition, ligand-based nuclear receptor profiling of a set of 2944 drugs provided an alert for 153 drugs. For some of them, namely, acitretin, telmisartan, phenyltoloxamine, tazarotene, and flumazenil, bibliographical evidence could be found indicating that those drugs may indeed have some potential off-target residual affinity for the nuclear receptors annotated. Overall, the present findings suggest that ligand-based approaches to protein family profiling appear as a promising means toward the establishment of novel tools for in silico pharmacology. PMID- 17125213 TI - 3D QSAR selectivity analyses of carbonic anhydrase inhibitors: insights for the design of isozyme selective inhibitors. AB - A 3D QSAR selectivity analysis of carbonic anhydrase (CA) inhibitors using a data set of 87 CA inhibitors is reported. After ligand minimization in the binding pockets of CA I, CA II, and CA IV isoforms, selectivity CoMFA and CoMSIA 3D QSAR models have been derived by taking the affinity differences (DeltapKi) with respect to two CA isozymes as independent variables. Evaluation of the developed 3D QSAR selectivity models allows us to determine amino acids in the respective CA isozymes that possibly play a crucial role for selective inhibition of these isozymes. We further combined the ligand-based 3D QSAR models with the docking program AUTODOCK in order to screen for novel CA inhibitors. Correct binding modes are predicted for various CA inhibitors with respect to known crystal structures. Furthermore, in combination with the developed 3D QSAR models we could successfully estimate the affinity of CA inhibitors even in cases where the applied scoring function failed. This novel strategy to combine AUTODOCK poses with CoMFA/CoMSIA 3D QSAR models can be used as a guideline to assess the relevance of generated binding modes and to accurately predict the binding affinity of newly designed CA inhibitors that could play a crucial role in the treatment of pathologies such as tumors, obesity, or glaucoma. PMID- 17125216 TI - Chiral decalins: preparation from oleanolic acid and application in the synthesis of (-)-9-epi-ambrox. AB - A novel and versatile process was developed to prepare the trans-decalins Delta9(11)-3beta-acetoxysclareolide (2) and Delta9(11)-3beta-acetoxy-8-epi sclareolide (3), respectively, with 4a-methoxycarbonyl-2,7,7-trimethyl-1-oxo-cis decalin-2-ene (4) and its C-3 hydroxyl derivative 5 from oleanolic acid (3beta hydroxyolean-12-en-28-oic acid, 1). Three key steps were (a) introduction of the AcO-12 group and the C-9,C-11 double bond at ring C of methyl 3beta-acetoxyolean 12-en-28-oate (8) to afford the diene, methyl 3,12-diacetoxyolean-9(11),12-dien 28-oate (11); (b) photolytic cleavage of the C-8,C-14 bond in the diene to give an acetoxy-substituted triene 14; and (c) oxidative cleavage of the triene or its hydrolyzed alpha,beta-unsaturated ketone product with m-CPBA/TsOH to give the cis and trans-decalins 2-5. Delata9(11)-3beta-Acetoxysclareolide (2) was stereospecifically reduced to give 3beta-acetoxy-9-epi-sclareolide (23), from which (-)-9-epi-ambrox (7) was synthesized. PMID- 17125217 TI - Trungapeptins A-C, cyclodepsipeptides from the marine cyanobacterium Lyngbya majuscula. AB - Trungapeptins A-C (1-3) were isolated from the marine cyanobacterium Lyngbya majuscula collected from Trung Province, Thailand. Their gross structures were elucidated by interpretation of spectroscopic data. The absolute configurations of the amino acids and phenyllactic acid were determined by Marfey's and chiral HPLC analyses, respectively. The relative stereochemistry of 3-hydroxy-2-methyl-7 octynoic acid (Hmoya) of trungapeptin A was elucidated by application of the J based configuration analysis, and its absolute stereochemistry was established to be 2S, 3R by Mosher's method. The structures of compounds 1-3 are closely related to the antanapeptins, a series of depsipeptides isolated from a Madagascan collection of L. majuscula. PMID- 17125218 TI - Cytotoxic triterpenoids from the leaves of Microtropis fokienensis. AB - Five new triterpenoids, microfokienoxanes A-D (1-4) and 3beta,28-dihydroxy 11alpha-methoxyurs-12-ene (5), were isolated and identified from the leaves of Microtropis fokienensis, along with nine known compounds. The structures of the new compounds were elucidated by spectroscopic methods. The compounds obtained in this investigation were evaluated against a small panel of human cancer cell lines for cytotoxicity. Only compounds 3 and 5 exhibited cytotoxicity (IC50 < or = 5 microg/mL) for one or more cell lines. PMID- 17125219 TI - Kahalalide derivatives from the Indian sacoglossan mollusk Elysia grandifolia. AB - Two new cyclic depsipeptide derivatives, kahalalides R (1) and S (2), together with two known congeners, kahalalides F (3) and D (4), were isolated from the Indian sacoglossan mollusk Elysia grandifolia. The structures of the new compounds were unambiguously established on the basis of NMR spectroscopic (1H, 13C, COSY, HMBC) and mass spectrometric (FABMS, ESIMS, MALDI-TOF/PSD) data, which also included Marfey amino acid analyses. The new derivative kahalalide R was found to exert comparable or even higher cytotoxicity than the potential drug candidate kahalalide F toward the MCF7 human mammary carcinoma cell line. PMID- 17125220 TI - Crassolides A-F, cembranoids with a trans-fused lactone from the soft coral Sarcophyton crassocaule. AB - Six new polyoxygenated cembrane-based diterpenoids possessing a trans-fused alpha methylene-gamma-lactone, crassocolides A-F (1-6), have been isolated along with the known compound 7 from the ethyl acetate extract of a Taiwanese soft coral, Sarcophyton crassocaule. The structures of 1-6 have been established by detailed spectroscopic analysis, including 2D NMR (1H-1H COSY, HMQC, HMBC, and NOESY) spectroscopy, while their absolute configurations were determined using a modified Mosher's method for 1. The structure of 5 was further proven by X-ray diffraction analysis. The full assignment of NMR data of 7 is reported herein for the first time. The cytotoxicity of crassocolides 1-4, 6, and 7 against a limited panel of cancer cells was also determined. PMID- 17125221 TI - A chemical study of cyclic depsipeptides produced by a sponge-derived fungus. AB - Two novel cyclic depsipeptides, guangomides A (1) and B (2), together with a new destruxin derivative (3) were isolated from the cytotoxic extract obtained from the saltwater culture of an unidentifiable sponge-derived fungus. The new structures were elucidated on the basis of analysis of extensive 1D and 2D NMR data sets, and the absolute configurations of 2S, 9S, 13S, 19S, 24R, 28R of 1 were determined on the basis of the combined X-ray and Marfey's method structure analysis. Identical absolute configurations were assumed for 2. The cytotoxicity of the extract was found to be due to brefeldin A, while 1 and 2 showed weak antibacterial activity against Staphylococcus epidermidis and Enterococcus durans. PMID- 17125222 TI - Sesquiterpenes from Thapsia nitida var. meridionalis and Thapsia nitida var. nitida. AB - Nine new eudesmanolides (1-9), two new guaianolides (12 and 13), and a new germacrane (10), along with a previously reported guaianolide (11), have been isolated from the roots of Thapsia nitida var. meridionalis. Thapsia nitida var. nitida also afforded compound 13 along with a new guaianolide (14). The structure of 13 was confirmed by X-ray crystallographic analysis. Compounds 1, 2, and 11-14 have been tested as potential inhibitors of the sarco- and endoplasmic Ca2+ dependent ATPases (SERCA) pump. None of them showed significant activities. PMID- 17125223 TI - Protein tyrosine phosphatase-1B inhibitory activity of isoprenylated flavonoids isolated from Erythrina mildbraedii. AB - Inhibition of protein tyrosine phosphatase-1B (PTP1B) has been proposed as a therapy for treatment of type-2 diabetes and obesity. Bioassay-guided fractionation of an EtOAc-soluble extract of the root bark of Erythrina mildbraedii, using an in vitro PTP1B inhibitory assay, resulted in the isolation of three new isoprenylated flavonoids, abyssinone-IV-4'-O-methyl ether (2), 7 hydroxy-4'-methoxy-3'-(3-hydroxy-3-methyl-trans-but-1-enyl)-5'-(3-methylbut-2 enyl)flavanone (3), and abyssinone-VI-4-O-methyl ether (6), along with six known flavonoids, abyssinone-V-4'-O-methyl ether (1), abyssinone-V (4), abyssinone-IV (5), sigmoidin E (7), 4'-hydroxy-5,7-dimethoxyisoflavone (8), and alpinumisoflavone (9). Compounds 1 and 2, 4-7, and 9 inhibited PTP1B activity, with IC50 values ranging from 14.8 +/- 1.1 to 39.7 +/- 2.5 microM. On the basis of the data obtained, flavanones and chalcones with isoprenyl groups may be considered as a new class of PTP1B inhibitors. PMID- 17125224 TI - Biologically active triterpenoid saponins from Acanthopanax senticosus. AB - Three new triterpenoid saponins, acanthopanaxosides A (1), B (7), and C (13), were isolated from the leaves of Acanthopanax senticosus, together with 12 known saponins. The structures of these new saponins were established as 3-O-beta-D glucopyranosyl-(1-->2)-alpha-L-arabinopyranosyl-30-nor-olean-12,20(29)-dien-28 oic acid 28-O-alpha-L-rhamnopyranosyl-(1-->4)-6-O-acetyl-beta-D-glucopyranosyl-(1 ->6)-beta-D-glucopyranosyl ester (1), 3-O-beta-D-glucopyranosyl-(1-->2)-alpha-L arabinopyranosyl oleanolic acid 28-O-alpha-L-rhamnopyranosyl-(1-->4)-6-O-acetyl beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester (7), and 3-O-alpha-L rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl-3beta-hydroxyolean-12-ene-28,29 dioic acid (13), on the basis of spectroscopic analysis and chemical degradation. Among the known compounds, sessiloside and tauroside H1 are reported for the first time from A. senticosus. The biological activity of compounds 1-15 was examined against pancreatic lipase. Ciwujianoside C1 (6), tauroside H1 (11), 3-O alpha-rhamnopyranosyl-(1-->2)-alpha-arabinopyranosyl mesembryanthemoidigenic acid (12), acanthopanaxoside C (13), sessiloside (14), and chiisanoside (15) inhibited pancreatic lipase activity in vitro. In turn, ciwujianosides C2 (3), D2 (5), C4 (8), and C3 (10) and hederasaponin B (9) enhanced this enzyme. PMID- 17125225 TI - Theopapuamide, a cyclic depsipeptide from a Papua New Guinea lithistid sponge Theonella swinhoei. AB - Theopapuamide (1), a new cytotoxic peptide, has been isolated from the lithistid sponge Theonella swinhoei from Papua New Guinea. The structure was established by analysis of NMR, mass spectrometry, and chemical methods. The undecapeptide (1) contains several unusual amino acid residues, of which the occurrence of beta methoxyasparagine and 4-amino-5-methyl-2,3,5-trihydroxyhexanoic acid (Amtha) is unprecedented in natural peptides. Compound 1 also contains an amide-linked fatty acid moiety, 3-hydroxy-2,4,6-trimethyloctanoic acid (Htoa). Theopapuamide (1) was cytotoxic against CEM-TART and HCT-116 cell lines, with EC50 values of 0.5 and 0.9 microM, respectively. PMID- 17125226 TI - Mooloolabenes A-E, norsesterterpenes from the Australian sponge Hyattella intestinalis. AB - Five new norsesterterpenes, mooloolabenes A-E (1-5), and the new sesterterpene mooloolaldehyde (6), related to the scalarane family of compounds, were isolated from an acetone extract of the Australian sponge Hyattella intestinalis. Structural elucidation, including relative stereochemical assignment, was based on spectroscopic analysis. All compounds tested showed cytotoxic activity against the P388 cell line. PMID- 17125227 TI - Triterpene saponins from clematis mandshurica. AB - Four new triterpene saponins, clematomandshurica saponins A-D (1-4), together with three known saponins (5-7) have been isolated from the roots and rhizomes of Clematis mandshurica. Their structures were elucidated on the basis of their spectroscopic evidence and hydrolysis. Clematomandshurica saponins A and B showed significant inhibitory activity on cyclooxygenase-2 (IC50 = 2.66 and 2.58 microM, respectively). PMID- 17125228 TI - Synthesis of bromoindole alkaloids from Laurencia brongniartii. AB - A regioselective synthesis of N-carbomethoxy-2,3,5-tribromoindole (6) via a sequential one-pot bromination-aromatization-bromination of N carbomethoxyindoline (2) is described. The process for the transformation of 2 into 6 permitted the isolation of stable reaction intermediates N-carbomethoxy-5 bromoindoline (3), N-carbomethoxy-5-bromoindole (4), and N-carbomethoxy-3,5 dibromoindole (5). Compound 6 was used to complete the total synthesis of the natural products 1b and 1c. In addition, bromination of N-carbomethoxyindole (11) afforded N-carbomethoxy-2,3,6-tribromoindole (13), from which the natural product 1a was synthesized. PMID- 17125229 TI - Penifulvins B-E and a silphinene analogue: sesquiterpenoids from a fungicolous isolate of Penicillium griseofulvum. AB - Penifulvins B-E (2-5), four new sesquiterpenoids with a dioxa[5.5.5.6]fenestrane ring system, have been isolated from cultures of an isolate of Penicillium griseofulvum (NRRL 35584), together with a new silphinene derivative, 12 hydroxysilphinene-15-oic acid (6). Penifulvins B-E (2-5) are oxidized analogues of penifulvin A (1) and were identified by analysis of NMR and MS data. 12 Hydroxysilphinen-15-oic acid (6) is biogenetically similar, and penifulvins A-E are presumed to be derived from a silphinene precursor. The structures of 2-6, including absolute configuration, were assigned by analysis of NMR data and application of chemical methods. PMID- 17125230 TI - Oleanane and taraxerane glycosides from the roots of Gomphrena macrocephala. AB - Phytochemical screening of the roots of Gomphrena macrocephala, with particular attention to its triterpene glycoside constituents, has resulted in the isolation of two new oleanane glycosides (1 and 2) and a new taraxerane glycoside (3). The structures of 1-3 were determined as 11alpha,12alpha-epoxy-3beta-[(O-beta-D glucuronopyranosyl)oxy]olean-28,13-olide (1), 11alpha,12alpha-epoxy-3beta-[(O beta-D-galactopyranosyl-(1-->3)-O-[beta-D-glucopyranosyl-(1-->2)]-beta-D glucuronopyranosyl)-oxy]olean-28,13-olide (2), and 11alpha,12alpha-epoxy-3beta [(O-beta-D-glucuronopyranosyl)oxy]taraxer-14-en-28-oic acid beta-D-glucopyranosyl ester (3), respectively, on the basis of their spectroscopic data and the results of hydrolysis. The aglycones (1a and 3a) of 1-3 with an epoxy group showed cytotoxic activity against HSC-2 human oral squamous carcinoma cells. PMID- 17125231 TI - C35 terpenoids from the bark of Calocedrus macrolepis var. formosana with activity against human cancer cell lines. AB - Ferrugicadinol (1) and ferrugieudesmol (2), two new compounds with a unique C35 terpene skeleton, were isolated from the bark of Calocedrus macrolepis var. formosana. Their structures were elucidated principally from spectroscopic evidence. The two new C35 terpenes showed cytotoxicity against human oral epidermoid carcinoma KB cells with IC50 values ranging from 11 to 14 microM. PMID- 17125232 TI - Monolaterol, the first configurationally assigned phenylphenalenone derivative with a stereogenic center at C-9, from Monochoria elata. AB - Phytochemical analysis of the roots of Monochoria elata resulted in the structure elucidation of monolaterol, the first configurationally assigned phenylphenalenone-type natural product with a stereogenic center at the phenyl bearing carbon, C-9, and four known phenylphenalenones by MS and NMR methods. The absolute configuration of the new phenyldihydrophenalenediol was assigned by comparing the results of quantum chemical CD calculations and experimental CD spectra, and the crystal structure was determined by X-ray diffraction analysis. PMID- 17125233 TI - Cytotoxic cis-fused bicyclic sesquiterpenoids from Jatropha neopauciflora. AB - Analysis of polar fractions of the bark extract of Jatropha neopauciflora provided two uncommon cis-fused bicyclic sesquiterpenoids, which were characterized as (1R,2R,5S,6S,7S,10S)*-5-epi-eudesm-4(15)-ene-1alpha,2beta,6alpha triol (1) and (1R,2R,5S,6S,7R,10S)*-ax-4(15)-ene-1alpha,2beta,7beta-triol (2). Their absolute configuration and biogenesis were derived by correlations with congeners of known absolute configurations. Biological investigation of less polar fractions of the bark extract led to the isolation of moderately cytotoxic triterpenes, calenduladiol (3) and (3beta,16beta)-16-hydroxylup-20(29)-en-3-yl (E)-3-(4-hydroxyphenyl)prop-2-enoate (4). PMID- 17125234 TI - Chaetopyranin, a benzaldehyde derivative, and other related metabolites from Chaetomium globosum, an endophytic fungus derived from the marine red alga Polysiphonia urceolata. AB - Cultivation of the endophytic fungus Chaetomium globosum, which was isolated from the inner tissue of the marine red alga Polysiphonia urceolata, resulted in the isolation of chaetopyranin (1), a new benzaldehyde secondary metabolite. Ten known compounds were also isolated, including two benzaldehyde congeners, 2-(2',3 epoxy-1',3'-heptadienyl)-6-hydroxy-5-(3-methyl-2-butenyl)benzaldehyde (2) and isotetrahydroauroglaucin (3), two anthraquinone derivatives, erythroglaucin (4) and parietin (5), five asperentin derivatives including asperentin (6, also known as cladosporin), 5'-hydroxy-asperentin-8-methylether (7), asperentin-8-methyl ether (8), 4'-hydroxyasperentin (9), and 5'-hydroxyasperentin (10), and the prenylated diketopiperazine congener neoechinulin A (11). The structures of these compounds were determined on the basis of their spectroscopic data analysis (1H, 13C, 1H-1H COSY, HMQC, and HMBC NMR, as well as low- and high-resolution mass experiments). To our knowledge, compound 1 represents the first example of a 2H benzopyran derivative of marine algal-derived fungi as well as of the fungal genus Chaetomium. Each isolate was tested for its DPPH (1,1-diphenyl-2 picrylhydrazyl) radical-scavenging property. Compounds 1-4 were found to have moderate activity. Chaetopyranin (1) also exhibited moderate to weak cytotoxic activity toward several tumor cell lines. PMID- 17125235 TI - Bohemamines from a marine-derived Streptomyces sp. AB - Investigation of the culture extracts of a marine-derived Streptomyces sp. led to the isolation of three new bohemamine-type pyrrolizidine alkaloids, bohemamine B (1), bohemamine C (2), and 5-chlorobohemamine C (3). The structures were elucidated using NMR methods, and the relative stereochemistry was determined using double-pulsed-field-gradient spin echo (DPFGSE) NOE studies. PMID- 17125236 TI - A nitrogen-containing 3-alkyl-1,4-benzoquinone and a gomphilactone derivative from Embelia ribes. AB - An unusual nitrogen-containing 3-alkyl-1,4-benzoquinone derivative, N-(3 carboxylpropyl)-5-amino-2-hydroxy-3-tridecyl-1,4-benzoquinone (1), and a gomphilactone derivative, 5,6-dihydroxy-7-tridecyl-3-[4-tridecyl-3-hydroxy-5-oxo 2(5H)-furylidene]-2-oxo-3(2H)-benzofuran (2), together with 14 known compounds, as well as the common plant metabolites sitosterol and daucosterol, were isolated from the ethanolic extract of the roots of Embelia ribes. Their structures were elucidated by means of spectroscopic methods. Cytotoxicities of the purified compounds were tested. PMID- 17125237 TI - Heterologous biosynthesis of amidated polyketides with novel cyclization regioselectivity from oxytetracycline polyketide synthase. AB - Heterologous expression of the minimal oxytetracycline polyketide synthase and the amidotransferase OxyD in Streptomyces coelicolor strain CH999 afforded two novel amidated polyketides, WJ85 (4) and WJ85b (5). WJ85 is a C-9 unreduced decaketide that is primed by a malonamyl starter unit. WJ85 is cyclized via an unusual C-11 to C-16 intramolecular aldol condensation not observed among known aromatic polyketides. The structures of WJ85 and the previously characterized WJ35 suggest that the presence of an amide starter unit has a profound effect on the cyclization regioselectivity and reactivity of a polyketide backbone. WJ85b is an anthraquinone and is an oxidized product of WJ85. PMID- 17125238 TI - Terpenoids and a diarylheptanoid from Zingiber ottensii. AB - Four new terpenoids and a diarylheptanoid were isolated together with 16 known compounds from rhizomes of Zingiber ottensii. The structures of the new compounds were determined to be 1,10,10-trimethylbicyclo[7,4,0]tridecane-3,6-dione (1), (E) 14-hydroxy-15-norlabda-8(17),12-dien-16-al (2), (E)-labda-8(17),12,14-trien 15(16)-olide (3), (E)-14,15,16-trinorlabda-8(17),11-dien-13-oic acid (4), and rel (3R,5S)-3,5-dihydroxy-1-(4-hydroxy-3-methoxyphenyl)-7-(3, 4 dihydroxyphenyl)heptane (5) by spectroscopic evidence. PMID- 17125239 TI - Acetylated flavanone glycosides from the rhizomes of Cyclosorus acuminatus. AB - Six new flavanone glycosides (1-6) were isolated from the methanol extract of the rhizomes of Cyclosorus acuminatus, together with the parent flavanone glycoside 2a. Their structures were established on the basis of spectroscopic and chemical methods. All compounds showed moderate activity against Streptococcus pneumoniae and Haemophilus influenzae. PMID- 17125240 TI - Acylphloroglucinol derivatives from Hypericum prolificum. AB - Three new acylphloroglucinol derivatives have been isolated from the hexane extract of the aerial parts of Hypericum prolificum L.: prolificin A (1), prolifenone A (2), and prolifenone B (3). The structures were elucidated on the basis of extensive 2D NMR and MS data. All three compounds were evaluated for in vitro cell proliferation inhibitory activity against human breast (MCF-7), lung (NCI-H460), CNS (SF-268), stomach (AGS), and colon (HCT-116) tumor cell lines. Prolificin A showed growth inhibition of all cell lines with IC50 values ranging from 23 to 36 microM. Prolifenones A and B were inactive at the concentrations tested. PMID- 17125241 TI - Prenylated flavonoids from the root bark of Berchemia discolor, a Tanzanian medicinal plant. AB - Five new prenylated flavonoids (1-5) were isolated from the root bark of Berchemia discolor, collected in Tanzania, along with 10 known compounds, by bioactivity-guided fractionation. The structures of compounds 1-5 were elucidated using various spectroscopic techniques. Of these isolates, compound 4, and the known compounds nitidulin (6), amorphigenin (7), and dabinol (8), exhibited cytotoxic activity when evaluated against a small panel of human cancer cells. Nitidulin (6) was further tested in an in vivo hollow fiber assay and found to be active against LNCaP (human hormone-dependent prostate cancer) cells implanted intraperitoneally, at doses of 10, 20, and 40 mg/kg. PMID- 17125242 TI - Comparative assessment of technologies for extraction of artemisinin. AB - This paper describes results of a multiobjective comparative assessment of several established and emerging technologies for extraction of a natural antimalarial substance, artemisinin. Extractions by hexane, supercritical carbon dioxide, hydrofluorocarbon HFC-134a, ionic liquids, and ethanol were considered. Hexane extraction is an established technology and appears to be the most cost effective. However, it is characterized by lower rates and efficiency of extraction than all other considered techniques and is also worse in terms of safety and environmental impact. Similarly, EtOH extraction was found to be worse than hexane in all assessment parameters. The new technologies (scCO2, HFC, and ILs) are based on nonflammable solvents and are characterized by faster extraction cycles and more complete extraction of the useful substances and enable continuous extraction processes with reduced solvent inventory. Ionic liquid and HFC-134a technologies show considerable promise and should be able to compete with hexane extraction in terms of cost-effectiveness following due process optimization. New technologies are also considerably safer (no risk of explosions, low toxicity) and greener (having a lower environmental impact in use, potential for biodegradability after use). The methodology of comparative assessment of established and emerging technologies is discussed. PMID- 17125245 TI - Acyl glucuronides: biological activity, chemical reactivity, and chemical synthesis. PMID- 17125246 TI - Discovery and development of 5-[(5S,9R)-9-(4-cyanophenyl)-3-(3,5-dichlorophenyl) 1-methyl-2,4-dioxo-1,3,7-triazaspiro[4.4]non-7-yl-methyl]-3-thiophenecarboxylic acid (BMS-587101)--a small molecule antagonist of leukocyte function associated antigen-1. AB - LFA-1 (leukocyte function-associated antigen-1), is a member of the beta2 integrin family and is expressed on all leukocytes. This letter describes the discovery and preliminary SAR of spirocyclic hydantoin based LFA-1 antagonists that culminated in the identification of analog 8 as a clinical candidate. We also report the first example of the efficacy of a small molecule LFA-1 antagonist in combination with CTLA-4Ig in an animal model of transplant rejection. PMID- 17125247 TI - Discovery of conformationally constrained tetracyclic compounds as potent hepatitis C virus NS5B RNA polymerase inhibitors. AB - We report a new series of hepatitis C virus NS5B RNA polymerase inhibitors containing a conformationally constrained tetracyclic scaffold. SAR studies led to the identification of 6,7-dihydro-5H-benzo[5,6][1,4]diazepino[7,1-a]indoles (19 and 20) bearing a basic pendent group with high biochemical and cellular potencies. These compounds displayed a very small shift in cellular potency when the replicon assay was performed in the presence of human serum albumin. PMID- 17125248 TI - Design and synthesis of novel isoquinoline-3-nitriles as orally bioavailable Kv1.5 antagonists for the treatment of atrial fibrillation. AB - Novel 3-cyanoisoquinoline Kv1.5 antagonists have been prepared and evaluated in in vitro and in vivo assays for inhibition of the Kv1.5 potassium channel and its associated cardiac potassium current, IKur. Structural modifications of isoquinolinone lead 1 afforded compounds with excellent potency, selectivity, and oral bioavailability. PMID- 17125249 TI - Estimation of volume of distribution in humans from high throughput HPLC-based measurements of human serum albumin binding and immobilized artificial membrane partitioning. AB - The volume of distribution (VD) in humans of 179 known drug molecules (acids, bases, and neutrals) has been modeled using two biomimetic-binding measurements. The phospholipid binding (log K (IAM)) and the plasma protein binding (log K (HSA)) have been calculated from gradient HPLC retention times on immobilized artificial membrane (IAM) and on human serum albumin (HSA) columns, respectively. The log VD values showed good correlation with the compounds' relative binding to IAM and HSA as follows: log VD=0.44 log K (IAM)-0.22 log K (HSA)-0.66; n=179, r2=0.76, s=0.33, and F=272. It was also observed that positively charged molecules bind relatively more to IAM, while negatively charged ones bind more to HSA, in line with the empirical observation that bases tend to have a larger volume of distribution than acids. These results suggest that with the help of these two simple high throughput HPLC-based biomimetic binding measurements an important in vivo drug disposition property can be estimated for use in early drug discovery. PMID- 17125250 TI - Fragment-based drug design: how big is too big? AB - Much has been discussed about the proper physicochemical properties (e.g., molecular weight, hydrophobicity, etc.) that should be considered when utilizing fragment leads in drug design. However, little has been reported as to what emphasis, if any, should be placed on the potency of the resulting fragment leads. In this report, a retrospective analysis of 18 highly optimized inhibitors is described in which the compounds were systematically deconstructed until the minimal binding elements could be identified. An analysis of the potency changes that were observed as the leads were reduced in size indicate that a nearly linear relationship exists between molecular weight and binding affinity over the entire range of sizes and potencies represented in the dataset. On the basis of these observations, prediction maps can be constructed that enable critical and quantitative assessments of the process of lead identification and optimization. These data place well-defined limits on the ideal size and potency of fragment leads that are being considered for use in fragment-based drug design. PMID- 17125251 TI - A 2.13 A structure of E. coli dihydrofolate reductase bound to a novel competitive inhibitor reveals a new binding surface involving the M20 loop region. AB - Dihydrofolate reductase (DHFR) is a vital metabolic enzyme and thus a clinically prominent target in the design of antimetabolites. In this work, we identify 1,4 bis-{[N-(1-imino-1-guanidino-methyl)]sulfanylmethyl}-3,6-dimethyl-benzene (compound 1) as the correct structure of the previously reported DHFR inhibitor 1,4-bis-{(iminothioureidomethyl)aminomethyl}-3,6-dimethyl-benzene (compound 2). The fact that compound 1 has an uncharacteristic structure for DHFR inhibitors, and an affinity (KI of 11.5 nM) comparable to potent inhibitors such as methotrexate and trimethoprim, made this inhibitor of interest for further analysis. We have conducted a characterization of the primary interactions of compound 1 and DHFR using a combination of X-ray structure and SAR analysis. The crystal structure of E. coli DHFR in complex with compound 1 and NADPH reveals that one portion of this inhibitor exploits a unique binding surface, the M20 loop. The importance of this interface was further confirmed by SAR analysis and additional structural characterization. PMID- 17125252 TI - Synthetic inhibitors of cytochrome P-450 2A6: inhibitory activity, difference spectra, mechanism of inhibition, and protein cocrystallization. AB - A series of 3-heteroaromatic analogues of nicotine were synthesized to delineate structural and mechanistic requirements for selectively inhibiting human cytochrome P450 (CYP) 2A6. Thiophene, substituted thiophene, furan, substituted furan, acetylene, imidazole, substituted imidazole, thiazole, pyrazole, substituted pyrazole, and aliphatic and isoxazol moieties were used to replace the N-methylpyrrolidine ring of nicotine. A number of potent inhibitors were identified, and several exhibited high selectivity for CYP2A6 relative to CYP2E1, -3A4, -2B6, -2C9, -2C19, and -2D6. The majority of these inhibitors elicited type II difference spectra indicating the formation of a coordinate covalent bond to the heme iron. The majority of inhibitors were reversible inhibitors although several mechanism-based inactivators were identified. Most of the inhibitors were also relatively metabolically stable. X-ray crystal structures of CYP2A6 cocrystallized with three furan analogues bearing methanamino side chains indicated that the amine side chain coordinated to the heme iron. The pyridyl moiety was positioned to accept a hydrogen bond from Asn297, and all three inhibitors exhibited orthogonal aromatic-aromatic interactions with protein side chains. For comparison, the cocrystal structure of 4,4'-dipyridyl disulfide was also obtained and showed that the pyridine moiety could assume a different orientation than that observed for the 3-heteroaromatic pyridines examined. For the 3-heteroromatic pyridines, N-methyl and N,N-dimethyl amino groups increased the apparent Ki and distorted helix I of the protein. Substitution of a phenyl ring for the pyridyl ring also increased the apparent Ki, which is likely to reflect the loss of the hydrogen bonding interaction with Asn297. In contrast, inhibitory potency for other P450s was increased, and the selectivity of the phenyl analogues for CYP2A6 was decreased relative to the pyridyl compounds. The results suggest that inhibitors that compliment the active site features of CYP2A6 can exhibit significant selectivity for CYP2A6 relative to other human liver drug-metabolizing P450s. PMID- 17125253 TI - Design, synthesis, and preliminary evaluation of doxazolidine carbamates as prodrugs activated by carboxylesterases. AB - The synthesis and tumor cell growth inhibition by doxazolidine carbamate prodrugs are reported. The carbamates were designed for selective hydrolysis by one or more human carboxylesterases to release doxazolidine (Doxaz), the formaldehyde oxazolidine of doxorubicin that cross-links DNA to trigger cell death. Simple butyl and pentyl, but not ethyl, carbamate prodrugs inhibited the growth of cancer cells that overexpress carboxylesterase CES1 (hCE1) and CES2 (hiCE). Relative CES1 and CES2 expression levels were determined by reverse transcription of the respective mRNAs, followed by polymerase chain reaction amplification. More complex structures with a p-aminobenzyl alcohol (PABA) self-eliminating spacer showed better growth inhibition (IC50=50 nM for Hep G2 liver cancer cells) while exhibiting reduced toxicity toward rat cardiomyocytes, relative to the parent drug doxorubicin. Pentyl 4-(N doxazolidinylcarbonyloxymethyl)phenylcarbamate, the lead compound for further investigation, appears to be activated in Hep G2 cells that express both CES1 and CES2. PMID- 17125254 TI - Synthesis of DNA-directed pyrrolidinyl and piperidinyl confined alkylating chloroalkylaminoanthraquinones: potential for development of tumor-selective N oxides. AB - A novel series of 1,4-disubstituted chloroethylaminoanthraquinones, containing alkylating chloroethylamino functionalities as part of a rigid piperidinyl or pyrrolidinyl ring-system, have been prepared. The target compounds were prepared by ipso-displacement of halides of various anthraquinone chromophores by either hydroxylated or chlorinated piperidinyl- or pyrrolidinyl-alkylamino side chains. The chloroethylaminoanthraquinones were shown to alkylate guanine residues of linearized pBR322 (1-20 microM), and two symmetrically 1,4-disubstituted anthraquinones (compounds 14 and 15) were shown to interstrand cross-link DNA in the low nM range. Several 1,4-disubstituted chloroethylaminoanthraquinones were potently cytotoxic (IC50 values: 500-fold selective). The most potent and selective P2Y2 receptor agonist of the present series was 2-thio-UTP (EC50=50 nM, >or=30-fold selective vs P2Y4 and P2Y6). All modifications at the uracil base of UTP led to a decrease in potency at the P2Y4 receptor. A beta,gamma-dichloromethylene modification in the triphosphate chain of 5-bromo-UTP was tolerated by all three receptor subtypes, thus opening up a new strategy to obtain ectonucleotide diphosphohydrolase- and phosphatase resistant P2Y2, P2Y4, and P2Y6 receptor agonists. PMID- 17125261 TI - Endoperoxide derivatives from marine organisms: 1,2-dioxanes of the plakortin family as novel antimalarial agents. AB - Plakortin (1) is a remarkably simple 1,2-dioxane derivative, extracted from the marine sponge Plakortis simplex, showing a submicromolar activity against chloroquine-resistant strains of Plasmodium falciparum. Using plakortin as a novel antimalarial hit, we have prepared a series of semisynthetic derivatives in order to gain insights into the structural requirements of simple 1,2-dioxanes for exhibiting antimalarial activity. Their synthesis, spectroscopic and computational analysis, and in vitro antimalarial activity are herein reported. Results obtained, besides confirming the crucial role of the cycloperoxide functionality, revealed other structural features critical for antimalarial activity, namely the "Western" alkyl side chain, the dioxane ring conformation, and the absolute configuration of the stereogenic carbons on the 1,2-dioxane ring, when affecting the bioactive ring conformation. PMID- 17125262 TI - Potent, selective, and orally efficacious antagonists of melanin-concentrating hormone receptor 1. AB - The high expression of MCH in the hypothalamus with the lean hypophagic phenotype coupled with increased resting metabolic rate and resistance to high fat diet induced obesity of MCH KO mice has spurred considerable efforts to develop small molecule MCHR1 antagonists. Starting from a lead thienopyrimidinone series, structure-activity studies at the 3- and 6-positions of the thienopyrimidinone core afforded potent and selective MCHR1 antagonists with representative examples having suitable pharmacokinetic properties. Based on structure-activity relationships, a structural model for MCHR1 was constructed to explain the binding mode of these antagonists. In general, a good correlation was observed between pKas and activity in the right-hand side of the template, with Asp123 playing an important role in the enhancement of binding affinity. A representative example when evaluated chronically in diet-induced obese mice resulted in good weight loss effects. These antagonists provide a viable lead series in the discovery of new therapies for the treatment of obesity. PMID- 17125263 TI - 6-(4-chlorophenyl)-3-substituted-thieno[3,2-d]pyrimidin-4(3H)-one-based melanin concentrating hormone receptor 1 antagonist. AB - Genetic manipulation studies in mice at both the MCH receptor 1 (MCHR1) as well as the MCH peptide levels have implicated MCHR1 as a key player in energy homeostasis. The phenotype exhibited by these studies, that is, increased metabolic rate, resistance to high fat diet, and subsequent weight loss, has spurred considerable efforts to develop antagonists of MCHR1. In continuation of efforts directed toward this goal, the present work capitalizes on the putative binding mode of an MCH antagonist, resulting in the identification of several novel chemotypes that are potent and selective MCHR1 antagonists. In addition, the favorable pharmacokinetics of representative examples has allowed for the evaluation of an MCHR1 antagonist in a high fat diet-induced obese rodent model of obesity. The tolerability of the right-hand side of the template for diverse chemotypes accompanied by favorable effects on weight loss enhances the attractiveness of this template in the pursuit toward development of effective anti-obesity agents. PMID- 17125264 TI - Potent and orally bioavailable 8-bicyclo[2.2.2]octylxanthines as adenosine A1 receptor antagonists. AB - In the search for a selective adenosine A1 receptor antagonist with greater aqueous solubility than the compounds currently in clinical trials as diuretics, a series of 1,4-substituted 8-cyclohexyl and 8-bicyclo[2.2.2]octylxanthines were investigated. The binding affinities of a variety of cyclohexyl and bicyclo[2.2.2]octylxanthines for the rat and human adenosine A1, A2A, A2B, and A3 receptors are presented. Bicyclo[2.2.2]octylxanthine 16 exhibited good pharmaceutical properties and in vivo activity in a rat diuresis model (ED50=0.3 mg/kg po). Optimization of the bridgehead substituent led to propionic acid 29 (BG9928), which retained high potency (hA1, Ki=7 nM) and selectivity for the adenosine A1 receptor (915-fold versus adenosine A2A receptor; 12-fold versus adenosine A2B receptor) with improved oral efficacy in the rat diuresis model (ED50=0.01 mg/kg) as well as high oral bioavailability in rat, dog, and cynomolgus monkey. PMID- 17125265 TI - Tricyclic imidazoline derivatives as potent and selective adenosine A1 receptor antagonists. AB - Novel tricyclic imidazoline antagonists of the adenosine A1 receptor are described. For key compounds, the selectivity level over other adenosine receptor subtypes is examined along with their in vivo effects in a rat diuresis model. Compound 14, the (R)-isomer of 7,8-dihydro-8-ethyl-2-(4-bicyclo[2.2.2]octan-1-ol) 4-propyl-1H-imidazo[2,1-i]purin-5(4H)-one, is a particularly potent adenosine A1 receptor antagonist with good selectivity over the other three adenosine receptor subtypes: A1 (human) Ki=22 nM; A2A (human) Ki=4400 nM; A2B (human) Ki=580 nM; A3 (human) Ki>or=10,000 nM. Imidazoline 14 is a competitive adenosine A1 receptor antagonist with a pA2 value of 8.88 and is highly soluble in water (>100 mg/mL). In addition, it has an oral bioavailability of 84% and an oral half-life of 3.8 h in rats. When orally administered in a rat diuresis model, compound 14 promoted sodium excretion (ED50=0.01 mg/kg). This level of efficacy is comparable to that of BG9928, a selective adenosine A1 receptor antagonist that is currently in clinical trials as a treatment for congestive heart failure. Additional modifications to 14 also showed that the bridgehead hydroxyl group could be replaced with a propionic acid (compound 36) without a significant loss in binding affinity or in vivo activity. PMID- 17125266 TI - WB4101-related compounds: new, subtype-selective alpha1-adrenoreceptor antagonists (or inverse agonists?). AB - Our previous structure-affinity relationship study had considered the enantiomers of the naphthodioxane, tetrahydronaphthodioxane, and 2-methoxy-1-naphthoxy analogues (compounds 1, 3, and 2, respectively) of 2-(2,6 dimethoxyphenoxyethylaminomethyl)-1,4-benzodioxane, the well-known alpha1 adrenoceptor (alpha1-AR) antagonist WB4101, showing that such modifications significantly modulate the affinity and selectivity profile for alpha1-AR subtypes and 5-HT1A receptor. Here, we extend investigations to antagonist activity enclosing new enantiomeric pairs, namely those of the methoxytetrahydronaphthoxy and methoxybiphenyloxy WB4101 analogues (4 and 5-7, respectively) and of a double-modified WB4101 derivative (8) resulting from hybridization between 2 and 3. We found that (S)-2 is a very potent (pA2 10.68) and moderately selective alpha1D-AR antagonist and the hybrid (S)-8 is a potent (pA2 7.98) and highly selective alpha1A-AR antagonist. Both of these compounds and (S)-WB4101 seem to act as inverse agonists in a vascular model. The results, which generally validate the logic we followed in designing these eight compounds, are acceptably rationalized by comparative SAR analysis of binding and functional affinities. PMID- 17125267 TI - Conjugation of adenosine and hexa-(D-arginine) leads to a nanomolar bisubstrate analog inhibitor of basophilic protein kinases. AB - Conjugates of oligoarginine peptides with adenine, adenosine, adenosine-5' carboxylic acid, and 5-isoquinolinesulfonic acid were synthesized and characterized as bisubstrate-analog inhibitors of cAMP-dependent protein kinase. Adenosine and adenine derivatives were connected to the N- or C-terminus of peptides containing four to six L- or D-arginine residues via a linker with a length that had been optimized in structure-activity studies. The orientation of the peptide chain strongly affected the activity of compounds incorporating D arginines. The biligand inhibitor containing Hidaka's H9 isoquinolinesulfonamide connected to the L-peptide had 65 times higher potency than the corresponding adenosine-containing conjugate, while both types of the conjugate comprising D peptides had similar low nanomolar activity. Two of the most active adenosine- and H9-peptide conjugates were tested in the panel of 52 different kinases. At 1 microM concentration, both compounds showed strong (more than 95%) inhibition of several basophilic AGC kinases, including pharmaceutically important kinases ROCK II and PKB/Akt. PMID- 17125268 TI - Selective angiotensin II AT2 receptor agonists: arylbenzylimidazole structure activity relationships. AB - Structural alterations in the 2- and 5-positions of the first drug-like selective angiotensin II AT2 receptor agonist (1) have been performed. The imidazole ring system was proven to be a strong determinant for the AT2 selectivity, and with few exceptions all variations gave good AT2 receptor affinities and with retained high AT2/AT1 selectivities. On the contrary to the findings with AT1 receptor agonists, the impact of structural modifications in the 5-position of the AT2 selective compounds were less pronounced regarding activation of the AT2 receptor. The butyloxyphenyl (56) and the propylthienyl (50) derivatives were found to exert a high agonistic effect as deduced from their capacity to induce neurite elongation in neuronal cells, as does angiotensin II. PMID- 17125269 TI - QSAR modeling of human serum protein binding with several modeling techniques utilizing structure-information representation. AB - Four modeling techniques, using topological descriptors to represent molecular structure, were employed to produce models of human serum protein binding (% bound) on a data set of 1008 experimental values, carefully screened from publicly available sources. To our knowledge, this data is the largest set on human serum protein binding reported for QSAR modeling. The data was partitioned into a training set of 808 compounds and an external validation test set of 200 compounds. Partitioning was accomplished by clustering the compounds in a structure descriptor space so that random sampling of 20% of the whole data set produced an external test set that is a good representative of the training set with respect to both structure and protein binding values. The four modeling techniques include multiple linear regression (MLR), artificial neural networks (ANN), k-nearest neighbors (kNN), and support vector machines (SVM). With the exception of the MLR model, the ANN, kNN, and SVM QSARs were ensemble models. Training set correlation coefficients and mean absolute error ranged from r2=0.90 and MAE=7.6 for ANN to r2=0.61 and MAE=16.2 for MLR. Prediction results from the validation set yielded correlation coefficients and mean absolute errors which ranged from r2=0.70 and MAE=14.1 for ANN to a low of r2=0.59 and MAE=18.3 for the SVM model. Structure descriptors that contribute significantly to the models are discussed and compared with those found in other published models. For the ANN model, structure descriptor trends with respect to their affects on predicted protein binding can assist the chemist in structure modification during the drug design process. PMID- 17125270 TI - Substituted trans-stilbenes, including analogues of the natural product resveratrol, inhibit the human tumor necrosis factor alpha-induced activation of transcription factor nuclear factor kappaB. AB - The transcription factor nuclear factor kappaB (NF-kappaB), which regulates expression of numerous antiinflammatory genes as well as genes that promote development of the prosurvival, antiapoptotic state is up-regulated in many cancer cells. The natural product resveratrol, a polyphenolic trans-stilbene, has numerous biological activities and is a known inhibitor of activation of NF kappaB, which may account for some of its biological activities. Resveratrol exhibits activity against a wide variety of cancer cells and has demonstrated activity as a cancer chemopreventive against all stages, i.e., initiation, promotion, and progression. The biological activities of resveratrol are often ascribed to its antioxidant activity. Both antioxidant activity and biological activities of analogues of resveratrol depend upon the number and location of the hydroxy groups. In the present study, phenolic analogues of resveratrol and a series of substituted trans-stilbenes without hydroxy groups were compared with resveratrol for their abilities to inhibit the human tumor necrosis factor alpha induced (TNF-alpha) activation of NF-kappaB, using the Panomics NF-kappaB stable reporter cell line 293/NF-kappaB-luc. A series of 75 compounds was screened to identify substituted trans-stilbenes that were more active than resveratrol. Dose response studies of the most active compounds were carried out to obtain IC50 values. Numerous compounds were identified that were more active than resveratrol, including compounds that were devoid of hydroxy groups and were 100 fold more potent than resveratrol. The substituted trans-stilbenes that were potent inhibitors of the activation of NFkappaB generally did not exhibit antioxidant activity. The results from screening were confirmed using BV-2 microglial cells where resveratrol and analogues were shown to inhibit LPS induced COX-2 expression. PMID- 17125271 TI - Discovery of a new class of macrocyclic antagonists to the human motilin receptor. AB - A novel class of macrocyclic peptidomimetics was identified and optimized as potent antagonists to the human motilin receptor (hMOT-R). Well-defined structure activity relationships allowed for rapid optimization of potency that eventually led to high affinity antagonists to hMOT-R. Potency and antagonist functional activity were confirmed both in functional and cell-based assays, as well as on isolated rabbit intestinal smooth muscle strips. Rapid access to this novel class of macrocyclic target structures was made possible through two efficient and complementary solid-phase parallel synthetic approaches, both of which are reported herein. PMID- 17125272 TI - Synthesis and biological evaluation of new asymmetrical bisintercalators as potential antitumor drugs. AB - The good results obtained in the past decade with various types of potential bisintercalating agents, e.g., LU 79553, DMP 840, BisBFI, MCI3335, WMC-26, BisAC, BisPA, and the asymmetrical derivative WMC-79 (Chart 1), prompted us to investigate a new series of asymmetrical bisintercalators, compounds 1a-t (Chart 2), which can combine the potentiality of bisintercalation with a possible different mechanism of action due to two diverse chromophores. The DNA-binding properties of these compounds have been examined using fluorometric techniques: target compounds are excellent DNA ligands, with a clear preference for binding to AT-rich duplexes. In vitro cytotoxicity of these derivatives toward human hormone-refractory prostate adenocarcinoma cell line (PC-3) is described. Apoptosis assays of four selected compounds are also reported. Very potent cytotoxic compounds, some of them capable of inducing early apoptosis, have been identified. PMID- 17125273 TI - Synthesis and radioligand binding studies of methoxylated 1,2,3,4 tetrahydroisoquinolinium derivatives as ligands of the apamin-sensitive Ca2+ activated K+ channels. AB - Several methoxylated 1,2,3,4-tetrahydroisoquinoliniums derived from N-methyl laudanosine and N-methyl-noscapine were synthesized and evaluated for their affinity for apamin-sensitive binding sites. The quaternary ammonium derivatives have a higher affinity with regard to the tertiary amines. 6,7-Dimethoxy analogues possess a higher affinity than the 6,8- and 7,8-dimethoxy isomers. A 3,4-dimethoxybenzyl or a 2-naphthylmethyl moiety in C-1 position are more favorable than a 3,4-dimethoxyphenethyl group. Smaller groups such as propyl or isobutyl are unfavorable. In 6,7-dimethoxy analogues, increasing the size and lipophilicity with a naphthyl group in the C-1 position leads to a slight increase of affinity, while the same group in the 6,7,8-trimethoxy series is less favorable. The 6,7,8-trimethoxy derivative 3f is the first tertiary amine in the series to possess an affinity close to that of N-methyl-laudanosine and N-methyl noscapine. Moreover, electrophysiological studies show that the most effective compound 4f blocks the apamin-sensitive afterhyperpolarization in rat dopaminergic neurons. PMID- 17125274 TI - Application of phosphoramidate ProTide technology significantly improves antiviral potency of carbocyclic adenosine derivatives. AB - We report the application of phosphoramidate pronucleotide (ProTide) technology to the antiviral agent carbocyclic L-d4A (L-Cd4A). The phenyl methyl alaninyl parent ProTide of L-Cd4A was prepared by Grignard-mediated phosphorochloridate reaction and resulted in a compound with significantly improved anti-HIV (2600 fold) and HBV activity. We describe modifications of the aryl, ester, and amino acid regions of the ProTide and how these changes affect antiviral activity and metabolic stability. Separate and distinct SARs were noted for HIV and HBV. Additionally, ProTides were prepared from the D-nucleoside D-Cd4A and the dideoxy analogues L-CddA and D-CddA. These compounds showed more modest potency improvements over the parent drug. In conclusion, the ProTide approach is highly successful when applied to L-Cd4A with potency improvements in vitro as high as 9000-fold against HIV. With a view to preclinical candidate selection we carried out metabolic stability studies using cynomolgus monkey liver and intestinal S9 fractions. PMID- 17125275 TI - New orally active derivatives of artemisinin with high efficacy against multidrug resistant malaria in mice. AB - A new series of ether derivatives of dihydroartemisinin have been prepared and evaluated for their antimalarial activity against multidrug-resistant Plasmodium yoelii nigeriensis in mice by oral route. These new derivatives 7-17 are highly lipophilic (log P in the range of 5.51 to 7.19) as compared with beta-arteether (log P 3.84), and several of them are two- to four-fold more active than beta arteether. Among the ether derivatives, alpha-isomers are more active than the beta-isomers. The ether derivatives 12alpha and 14alpha, the most active compounds of the series, provided 100% protection to infected mice at 12 mg/kgx4 days. In this model beta-arteether provides 100% and 20% protection at 48 mg/kgx4 days and 24 mg/kgx4 days, respectively. PMID- 17125276 TI - Structure-activity relationships of a novel series of urotensin II analogues: identification of a urotensin II antagonist. AB - Urotensin II (U-II) is a potent vasoconstrictor peptide which has been identified as the endogenous ligand for the orphan G protein-coupled receptor GPR14 now renamed UT receptor. As the C-terminal cyclic hexapeptide of U-II (U-II(4-11), H Asp-Cys-Phe-Trp-Lys-Tyr-Cys-Val-OH) possesses full biological activity, we have synthesized a series of U-II(4-11) analogues and measured their binding affinity on hGPR14-transfected CHO cells and their contractile activity on de endothelialized rat aortic rings. The data indicate that a free amino group and a functionalized side-chain at the N-terminal extremity of the peptide are not required for biological activity. In addition, the minimal chemical requirement at position 9 of U-II(4-11) is the presence of an aromatic moiety. Most importantly, replacement of the Phe6 residue by cyclohexyl-Ala (Cha) led to an analogue, [Cha6]U-II(4-11), that was devoid of agonistic activity but was able to dose-dependently suppress the vasoconstrictor effect of U-II on rat aortic rings. These new pharmacological data, by providing further information regarding the structure-activity relationships of U-II analogues, should prove useful for the rational design of potent and nonpeptidic UT receptor agonists and antagonists. PMID- 17125277 TI - Discovering inhibitors of human sirtuin type 2: novel structural scaffolds. AB - A successful virtual screening experiment of novel SIRT2 inhibitors is described. Four out of 11 experimentally tested compounds showed in vitro inhibitory activity toward SIRT2 in a micromolar level, resulting in an experimental hit ratio of 36%. Two of these compounds inhibited SIRT2 with IC50 (microM) values of 51 and 91; moreover, one of the new inhibitors was comprised of an entirely new SIRT2-inhibiting structural scaffold. PMID- 17125278 TI - Novel Schiff base copper complexes of quinoline-2 carboxaldehyde as proteasome inhibitors in human prostate cancer cells. AB - We report the synthesis of novel 1:1 Schiff base copper complexes of quinoline-2 carboxaldehyde showing dose-dependent, antiproliferative, and proapoptotic activity in PC-3 and LNCaP prostate cancer cells. We found that quinoline thiosemicarbazone 2 (FPA-137) was the most potent and inhibited proteosome activity in intact human prostate cancer PC-3 and LNCaP cells (IC50 of 4 and 3.2 microM, respectively) compared to clioquinol and pyrrolidine dithiocarbamate (IC50 of 10 and 20 microM), supporting the novelty of 2. PMID- 17125279 TI - 1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazoles: identification of a potent Aurora kinase inhibitor with a favorable antitumor kinase inhibition profile. AB - The optimization of a series of 5-phenylacetyl 1,4,5,6-tetrahydropyrrolo[3,4 c]pyrazole derivatives toward the inhibition of Aurora kinases led to the identification of compound 9d. This is a potent inhibitor of Aurora kinases that also shows low nanomolar potency against additional anticancer kinase targets. Based on its high antiproliferative activity on different cancer cell lines, favorable chemico-physical and pharmacokinetic properties, and high efficacy in in vivo tumor models, compound 9d was ultimately selected for further development. PMID- 17125283 TI - Stochastic analysis of the parity-violating energy differences between enantiomers and its implications for the origin of biological chirality. AB - A stochastic description of a racemic mixture is developed taking into account the slight energy difference between enantiomers originating from parity violation (DeltaE(PV) approximately equal to 10(-13) Jmol(-1)). The system can be described by an asymmetric binomial distribution. A method is developed to calculate the probability of forming the more-stable isomer in excess, which is not significantly larger than 50% under normal conditions. It is concluded that the parity-violating energy difference between enantiomers is very unlikely to be relevant in considerations about the origin of biological chirality. PMID- 17125284 TI - Shedding light on biomolecule conformational dynamics using fluorescence measurements of trapped ions. AB - Biomolecule conformational change has been widely investigated in solution using several methods; however, much less experimental data about structural changes are available for completely isolated, gas-phase biomolecules. Studies of conformational change in unsolvated biomolecules are required to complement the interpretation of mass spectrometry measurements and in addition, can provide a means to directly test theoretical simulations of biomolecule structure and dynamics independent of a simulated solvent. In this Feature Article, we review our recent introduction of a fluorescence-based method for probing local conformational dynamics in unsolvated biomolecules through interactions of an attached dye with tryptophan (Trp) residues and fields originating on charge sites. Dye-derivatized biomolecule ions are formed by electrospray ionization and are trapped in a variable-temperature quadrupole ion trap in which they are irradiated with either continuous or short pulse lasers to excite fluorescence. Fluorescence is measured as a function of temperature for different charge states. Optical measurements of the dye fluorescence include average intensity changes, changes in the emission spectrum, and time-resolved measurements of the fluorescence decay. These measurements have been applied to the miniprotein, Trp cage, polyproline peptides and to a beta-hairpin-forming peptide, and the results are presented as examples of the broad applicability and utility of these methods. Model fits to Trp-cage fluorescence data measured as a function of temperature provide quantitative information on the thermodynamics of conformational changes, which are reproduced well by molecular dynamics. Time resolved measurements of the fluorescence decays of Trp-cage and small polyproline peptides definitively demonstrate the occurrence of fluorescence quenching by the amino acid Trp in unsolvated biomolecules. PMID- 17125285 TI - Ion yields of thin MALDI samples: dependence on matrix and metal substrate and implications for models. AB - Thin MALDI samples can perform differently than thicker samples, on metal substrates. Divergent results and models for the effect have been presented. Positive and negative yields are investigated here for three matrixes (2,5 dihydroxybenzoic acid (DHB), sinapinic acid, and alpha-cyano 4-hydroxycinnamic acid) on stainless steel and gold substrates. Samples were electrosprayed for uniformity and thickness control and imaged across a metal-metal boundary. Thin sample enhancement is found in both polarities for all three matrixes on a steel substrate. On gold, only alpha-cyano-4-hydroxycinnamic acid shows enhancement. These and earlier data are used to evaluate two models. The first is based on one photon photoelectron emission from the metal; the second one, on two-photon matrix ionization at the metal interface. The surface-enhanced matrix photoionization model best fits the evidence, including the fluence dependence of electron emission from DHB on steel. PMID- 17125286 TI - Photoinduced electron transfer and excitation energy transfer in directly linked zinc porphyrin/zinc phthalocyanine composite. AB - Photoinduced electron transfer (ET) and excitation energy transfer (ENT) reactions in monomer and slipped-cofacial dimer systems of a directly linked Zn porphyrin (Por)-Zn phthalocyanine (Pc) heterodyad, ZnPc-ZnPor, were investigated by means of the picosecond and femtosecond transient absorption spectroscopies. In the dimer dyad system of two heterodyads connected through the coordination bond between two imidazolyl-substituted ZnPor bearing ZnPc, ZnPc-ZnPor(D), the rapid ENT from the ZnPor to ZnPc in the subpicosecond time region was followed by photoinduced charge separation (CS) and charge recombination (CR) with time constants of 47 and 510 ps, respectively. On the other hand in the monomer dyad system, no clear charge-separated state was observed although the CS with a time constant of 200 ps and CR with < or =70 ps were estimated. These results indicated that the dimer slipped-cofacial arrangement of pair porphyrins is advantageous for the effective production of the CS state. This advantage was discussed from the viewpoint of a decrease in the reorganization energy of the dimer relative to that of the monomer system. In addition, the electrochemical measurements indicated that the strong interaction between ZnPc and ZnPor moieties also contributed to the fast CS process despite the marginal driving force for the CS process. The dimer dyad of ZnPc-ZnPor provides full advantages in efficiencies of the light harvesting and the CS state production. PMID- 17125287 TI - Proton-transfer reaction of 4-methyl 2,6-diformyl phenol in cyclodextrin nanocage. AB - We report here on the steady-state and time-resolved fluorescence studies on proton-transfer (PT) reaction of 4-methyl 2,6-diformyl phenol (MFOH) in confined nanocavities in three solvents, dimethyl sulfoxide (DMSO), dimethyl formamide (DMF), and water. Though DMSO and DMF individually interact with MFOH in a similar fashion, their modes of interaction get significantly modified in the presence of cyclodextrin (CD) nanocages. In DMSO, in the ground state, the solvated molecular anion of MFOH forms 1:1 inclusion complex with beta- or gamma CD and attains greater stability compared to the normal form. In DMF, the solvated molecular anion gets converted to the H-bonded complex within the CD cavity resulting in a 50-nm blue shift in the absorption spectra. In the excited state, the anionic species gets more stabilized in DMSO while in DMF it is significantly destabilized in the presence of CDs. However, in case of water, MFOH gets trapped inside the water cages so that the CDs fail to complex with it effectively. There are also no changes in the excited-state lifetimes in water in the presence of CDs, but in case of DMSO and DMF, because of restricted rotation of the formyl group within the CD cavity, the contribution of the shorter lifetime components reduce significantly increasing the larger components. Some theoretical calculations at the AM1 level of approximation have also been carried out to demonstrate how the dipolar nature of the solvent influences excited-state PT in confined media. PMID- 17125288 TI - Degenerate electron exchange reaction of n-alkane radical cations in solution. AB - The degenerate electron exchange (DEE) reaction involving radical cations (RCs) of n-nonane, n-dodecane, and n-hexadecane in n-hexane solution was studied over the temperature range 253-313 K using the method of time-resolved magnetic field effect in recombination fluorescence of spin-correlated radical ion pairs. In the dilute solutions the rate constant of DEE was found to be 200 times slower than the diffusion limit. Using n-nonane as an example, we showed that two reasons are responsible for the low value of the RC self-exchange rate: (1) conformational variability of molecules and RCs and (2) the activation barrier of DEE reaction. The calculations of the reaction enthalpy performed by the B3LYP/6-31G(d) method indicated that electron transfer can be effective only upon collision of RC with a neutral molecule either in the all-trans conformation or in the conformation differing from the latter by rotation of the end ethyl fragment. The activation barrier of the DEE reaction was estimated using the reorganization energy of the internal degrees of freedom calculated at the B3LYP level and was found to be about 6 kcal/mol. A possible influence of the interaction between RC and a neutral molecule in an encounter complex on DEE rate constant is also discussed. PMID- 17125289 TI - Intramolecular charge transfer and dielectric solvent relaxation in n-propyl cyanide. N-phenylpyrrole and 4-dimethylamino-4'-cyanostilbene. AB - Fast intramolecular charge transfer (ICT) accompanied by dual fluorescence from a locally excited (LE) and an ICT state taking place with N-phenylpyrrole (PP) in the solvent n-propyl cyanide (PrCN) is investigated as a function of temperature between 25 and -112 degrees C. The LE and ICT fluorescence decays from -45 to -70 degrees C can be adequately fitted with two exponentials, in accordance with a two state (LE + ICT) reaction mechanism, similar to what has been observed with PP in the more polar and less viscous alkyl cyanides acetonitrile (MeCN) and ethyl cyanide (EtCN). At lower temperatures, triple-exponential fits are required for the LE and ICT decays. The ICT emission band maximum of the time-resolved fluorescence spectra of PP in PrCN at -100 degrees C displays a spectral shift from 29 230 cm-1 at t = 0 to 27 780 cm-1 at infinite time, which equilibration process is attributed to dielectric solvent relaxation. From the time dependence of this shift, in global analysis with that of the band integrals BI(LE) and BI(ICT) of the time-resolved LE and ICT fluorescence bands, the decay times 119 and 456 ps are obtained. Dielectric relaxation times of 20 and 138 ps are determined from the double-exponential spectral solvation response function C(t) of the probe molecule 4-dimethylamino-4'-cyanostilbene in PrCN at -100 degrees C. It is concluded from the similarity of the times 119 ps (PP) and 138 ps (DCS) that the deviation from double-exponential character for the fluorescence decays of PP in PrCN below -70 degrees C is due to the interference of dielectric solvent relaxation with the ICT reaction. This fact complicates the kinetic analysis of the LE and ICT fluorescence decays. The kinetic analysis for PP in PrCN is hence restricted to temperatures between -70 and -45 degrees C. From this analysis, the forward and backward ICT activation energies Ea (12 kJ/mol) and Ed (17 kJ/mol) are obtained, giving an ICT stabilization enthalpy -DeltaH of 5 kJ/mol. A comparison of the reaction barriers for PP in the three alkyl cyanides PrCN, EtCN, and MeCN (J. Phys. Chem. A 2005, 109, 1497) shows that Ea becomes smaller with increasing solvent polarity (from 12 to 6 kJ/mol), whereas Ed remains effectively constant. Both observations are indicative of a late transition state for the LE --> ICT reaction. The significance of the Leffler Hammond postulate in this connection is discussed: not primarily the energy of the LE, ICT, and transition states but rather the extent of charge transfer in these states determines whether an early or a late transition state is present. PMID- 17125290 TI - Ultrafast structural dynamics of photochromic indolylfulgimides studied by vibrational spectroscopy and DFT calculations. AB - The structural dynamics of the ring-opening reaction in a photochromic indolylfulgimide, a reversible, ultrafast photoswitch, is investigated by ultra broadband time-resolved vibrational spectroscopy. The experimentally observed vibrational modes of the indolylfulgimide photoisomers C and E are assigned to normal modes with the help of DFT calculations. A complete evaluation of the observed vibrational dynamics including excited-state vibrational modes is used to characterize the reaction path and the cooling behavior of the photoswitch. PMID- 17125291 TI - Electron spectroscopy, molecular structures, and binding energies of Al- and Cu imidazole. AB - Al- and Cu-imidazole are produced in laser-vaporization supersonic molecular beams and studied with pulsed field ionization-zero electron kinetic energy (ZEKE) spectroscopy and second-order Moller-Plesset (MP2) theory. The sigma and pi structures of these complexes are predicted by MP2 calculations, but only the sigma structures are identified by the experimental measurements. For these sigma structures, adiabatic ionization energies and several vibrational frequencies are measured from the ZEKE spectra, the ground electronic states of the neutral and ionized complexes are determined by comparing the observed and calculated spectra, and the metal-ligand bond dissociation energies of the neutral states are derived by using a thermochemical relation. The measured vibrational modes include the metal-ligand stretch and bend and ligand ring distortions. The metal ligand stretch frequencies of these transient complexes are compared with those of coordinately saturated, stable metal compounds, and the ligand-based distortion frequencies are compared with those of the free ligand. Al-imidazole has a larger bond dissociation energy than Cu-imidazole, although the opposite order was previously found for the corresponding ions. The weaker bonding of the Cu complex is attributed to the antibonding interaction and the electron repulsion between the Cu 4s and N lone-pair electrons. PMID- 17125292 TI - Group 4 transition-metal atom reactions with CS2 and OCS: infrared spectra and density functional calculations of SMCS, SM-(eta2-CS), SMCO, and OMCS in solid argon. AB - Laser-ablated titanium, zirconium, and hafnium atoms were reacted with CS2 and OCS molecules during condensation in excess argon. With CS2, the SMCS and S M(eta2-CS) products were formed on sample deposition. Photolysis increased both complexes, while annealing favored the lower energy S-M(eta2-CS) side-bound isomer. The OCS reactions produced SMCO, OMCS, and the simple M(eta2-CO)S adduct. Product absorptions are identified by comparison with density functional theory frequency calculations and isotopic substitutions. Investigations with OCS emphasized differences in the CS and CO bond insertion products. PMID- 17125293 TI - Spectroscopic identification of carbenium and ammonium isomers of protonated aniline (AnH+): IR spectra of weakly bound AnH+ -Ln clusters (L = Ar, N2). AB - Infrared photodissociation (IRPD) spectra of mass-selected clusters composed of protonated aniline (C6H8N+ = AnH+) and a variable number of neutral ligands (L = Ar, N2) are obtained in the N-H stretch range. The AnH+ -Ln complexes (n < or = 3) are produced by chemical ionization in a supersonic expansion of An, H2, and L. The IRPD spectra of AnH+-Ln feature the unambiguous fingerprints of at least two different AnH+ nucleation centers, namely, the ammonium isomer (5) and the carbenium ions (1 and/or 3) corresponding to protonation at the N atom and at the C atoms in the para and/or ortho positions, respectively. Protonation at the meta and ipso positions is not observed. Both classes of observed AnH+-Ln isomers exhibit very different photofragmentation behavior upon vibrational excitation arising from the different interaction strengths of the AnH+ cores with the surrounding neutral ligands. Analysis of the incremental N-H stretch frequency shifts as a function of cluster size shows that microsolvation of both 5 and 1/3 in Ar and N2 starts with the formation of intermolecular H bonds of the ligands to the acidic NH protons and proceeds by intermolecular pi bonding to the aromatic ring. The analysis of both the photofragmentation branching ratios and the N-H stretch frequencies demonstrates that the N-H bonds in 5 are weaker and more acidic than those in 1/3, leading to stronger intermolecular H bonds with L. The interpretation of the spectroscopic data is supported by density functional calculations conducted at the B3LYP level using the 6-31G* and 6-311G(2df,2pd) basis sets. Comparison with clusters of neutral aniline and the aniline radical cation demonstrates the drastic effect of protonation and ionization on the acidity of the N-H bonds and the topology of the intermolecular potential, in particular on the preferred aromatic substrate-nonpolar ligand recognition motif. PMID- 17125294 TI - Proton transfer mediated by the vibronic coupling in oxygen core ionized states of glyoxalmonoxime studied by infrared-X-ray pump-probe spectroscopy. AB - The theory of IR-X-ray pump-probe spectroscopy beyond the Born-Oppenheimer approximation is developed and applied to the study of the dynamics of intramolecular proton transfer in glyoxalmonoxime leading to the formation of the tautomer 2-nitrosoethenol. Due to the IR pump pulses the molecule gains sufficient energy to promote a proton to a weakly bound well. A femtosecond X-ray pulse snapshots the wave packet route and, hence, the dynamics of the proton transfer. The glyoxalmonoxime molecule contains two chemically nonequivalent oxygen atoms that possess distinct roles in the hydrogen bond, a hydrogen donor and an acceptor. Core ionizations of these form two intersecting core-ionized states, the vibronic coupling between which along the OH stretching mode partially delocalizes the core hole, resulting in a hopping of the core hole from one site to another. This, in turn, affects the dynamics of the proton transfer in the core-ionized state. The quantum dynamical simulations of X-ray photoelectron spectra of glyoxalmonoxime driven by strong IR pulses demonstrate the general applicability of the technique for studies of intramolecular proton transfer in systems with vibronic coupling. PMID- 17125295 TI - Anion photoelectron spectroscopy and density functional investigation of vanadium carbide clusters. AB - The influence of source conditions on vanadium-carbon cluster formation in a methane-vanadium plasma is explored and analyzed by photoelectron spectroscopy, revealing that the metal-carbon ratio has substantial influence over the cluster products. Experiments that employ large methane content produce carbon-rich mono- and divanadium carbides. The carbon-rich clusters show a preference for the formation of cyclic neutral and linear ionic structures. When the methane concentration is decreased, VmCn clusters are formed with m = 1-4 and n = 2-8. The photoelectron spectra of clusters formed under these conditions are indicative of a three-dimensional network. We have measured a significantly lower vertical electron affinity for the VC2, V2C3, and V4C6 clusters compared with proximate species. Interestingly, the VC2 species is a proposed building block of the M8C12 Met-Car cluster, and the 2,3 and 4,6 clusters correspond to the 1/4 and 1/2 Met-Car cages, respectively. This correlation is taken as evidence of their importance in the formation of the larger Met-Car species. These results are supported by density functional theory (DFT) calculations carried out at the PBE/GGA level. PMID- 17125296 TI - Ring-expansion reactions in the thermal decomposition of tert-butyl-1,3 cyclopentadiene. AB - The thermal decomposition of tert-butyl-1,3-cyclopentadiene has been investigated in single-pulse shock-tube studies at shock pressures of 182-260 kPa and temperatures of 996-1127 K. Isobutene (2-methylpropene), 1,3-cyclopentadiene, and toluene were observed as the major stable products in the thermolysis of dilute mixtures of the substrate in the presence of a free-radical scavenger. Hydrogen atoms were also inferred to be a primary product of the decomposition and could be quantitatively determined on the basis of products derived from the free radical scavenger. Of particular interest is the formation of toluene, which involves the expansion of the ring from a five- to a six-membered system. The overall reaction mechanism is suggested to include isomerization of the starting material; a molecular elimination channel; and C-C bond fission reactions, with toluene formation occurring via radical intermediates formed in the latter pathway. These radical intermediates are analogous to those believed to be important in soot formation reactions occurring during combustion. Molecular and thermodynamic properties of key species were determined from G3MP2B3 quantum chemistry calculations and are reported. The temperature dependence of the product spectrum was fit with a detailed chemical kinetic model, and best-fit kinetic parameters were derived using a Nelder-Mead simplex minimization algorithm. Our mechanism and rate constants are consistent with and provide experimental support for the H-atom-assisted routes to the conversion of fulvene to benzene that have been proposed in the literature on the basis of theoretical investigations. PMID- 17125297 TI - Gas-phase reactions of atomic lanthanide cations with CO2 and CS2: room temperature kinetics and periodicities in reactivity. AB - Gas-phase reactions of atomic lanthanide cations (excluding Pm+) have been surveyed systematically with CO2 and CS2 using an inductively coupled plasma/selected-ion flow tube (ICP/SIFT) tandem mass spectrometer. Observations are reported for reactions with La+, Ce+, Pr+, Nd+, Sm+, Eu+, Gd+, Tb+, Dy+, Ho+, Er+, Tm+, Yb+, and Lu+ at room temperature (295 +/- 2 K) in helium at a total pressure of 0.35 +/- 0.02 Torr. The observed primary reaction channels correspond to X-atom transfer (X = O, S) and CX2 addition. X-atom transfer is the predominant reaction channel with La+, Ce+, Pr+, Nd+, Gd+, Tb+, and Lu+, and CX2 addition occurs with the other lanthanide cations. Competition between these two channels is seen only in the reactions of CS2 with Nd+ and Lu+. Rate coefficient measurements indicate a periodicity in the reaction efficiencies of the early and late lanthanides. With CO2 the observed trends in reactivity across the row and with exothermicity follow trends in the energy required to achieve two unpaired non-f valence electrons by electron promotion within the Ln+ cation that suggest the presence of a kinetic barrier, in a manner much like those observed previously for reactions with isoelectronic N2O. In contrast, no such barrier is evident for S-atom transfer from the valence isolectronic CS2 molecule which proceeds at unit efficiency, and this is attributed to the much higher polarizability of CS2 compared to CO2 and N2O. Up to five CX2 molecules were observed to add sequentially to selected Ln+ and LnX+ cations. PMID- 17125298 TI - Contribution to the chemistry of the Belousov-Zhabotinsky reaction. Products of the Ferriin-Bromomalonic acid and the Ferriin-Malonic acid reactions. AB - In the present mechanistic schemes of the ferroin-catalyzed oscillatory Belousov Zhabotinsky (BZ) reaction the oxidation of the organic substrates (bromomalonic or malonic acid) by ferriin (the oxidized form of the catalyst) plays an important role. As the organic products of these reactions were not yet identified experimentally, they were studied here by an HPLC technique. It was found that the main organic oxidation product of bromomalonic acid is bromo ethene-tricarboxylic acid (BrEETRA), the same compound that is formed when bromomalonic acid is oxidized by Ce4+ (another catalyst of the BZ reaction). Formation of BrEETRA is explained here by a new mechanism that is more realistic than the one suggested earlier. To find any oxidation product of malonic acid in the ferriin-malonic acid reaction was not successful, however. Neither ethane tetracarboxylic acid (ETA) nor malonyl malonate (MAMA), the usual products of the Ce4+- malonic acid reaction, nor any other organic acid, not even CO2, was found as a product of the reaction. We propose that malonic acid is not oxidized in the ferriin-malonic acid reaction, and it plays only the role of a complex forming catalyst in a process where Fe3+ oxidizes mostly its phenantroline ligand. PMID- 17125299 TI - Kinetics and mechanisms of CF3CHFOCH3, CF3CHFOC(O)H, and FC(O)OCH3 reactions with OH radicals. AB - The kinetics and mechanism of oxidation of CF3CHFOCH3 was studied using an 11.5 dm3 environmental reaction chamber. OH radicals were produced by UV photolysis of an O3-H2O-He mixture at an initial pressure of 200 Torr in the chamber. The rate constant of the reaction of CF3CHFOCH3 with OH radicals (k1) was determined to be (1.77 +/- 0.69) x 10(-12) exp[(-720 +/- 110)/T] cm3 molecule(-1)(s-1) by means of a relative rate method at 253-328 K. The mechanism of the reaction was investigated by FT-IR spectroscopy at 298 K. CF3CHFOC(O)H, FC(O)OCH3, and COF2 were determined to be the major products. The branching ratio (k1a/k1b) for the reactions CF3CHFOCH3 + OH --> CF3CHFOCH2* + H2O (k1a) and CF3CHFOCH3 + OH --> CF3CF*OCH3 + H2O (k1b) was estimated to be 4.2:1 at 298 K from the yields of CF3CHFOC(O)H, FC(O)OCH3, and COF2. The rate constants of the reactions of CF3CHFOC(O)H (k2) and FC(O)OCH3 (k3) with OH radicals were determined to be (9.14 +/- 2.78) x 10(-13) exp[(-1190 +/- 90)/T] and (2.10 +/- 0.65) x 10(-13) exp[(-630 +/- 90)/T] cm3 molecule(-1)(s-1), respectively, by means of a relative rate method at 253-328 K. The rate constants at 298 K were as follows: k1 = (1.56 +/- 0.06) x 10-13, k2 = (1.67 +/- 0.05) x 10-14, and k3 = (2.53 +/- 0.07) x 10-14 cm3 molecule(-1)(s-1). The tropospheric lifetimes of CF3CHFOCH3, CF3CHFOC(O)H, and FC(O)OCH3 with respect to reaction with OH radicals were estimated to be 0.29, 3.2, and 1.8 years, respectively. PMID- 17125300 TI - Absolute rate coefficient of the OH + CH(3)C(O)OH reaction at T = 287-802 K. The two faces of pre-reactive H-bonding. AB - The rate constants for the reaction OH + CH3C(O)OH --> products (1) were determined over the temperature range 287-802 K at 50 and 100 Torr of Ar or N2 bath gas using pulsed laser photolysis generation of OH by CH3C(O)OH photolysis at 193 nm coupled with OH detection by pulsed laser-induced fluorescence. The rate coefficient displays a complex temperature dependence with a sharp minimum at 530 K, indicating the competition between a reaction proceeding through a pre reactive H-bonded complex to form CH3C(O)O + H2O, expected to prevail at low temperatures, and a direct methyl-H abstraction channel leading to CH2C(O)OH + H2O, which should dominate at high temperatures. The temperature dependence of the rate constant can be described adequately by k1(287-802 K) = 2.9 x 10(-9) exp{-6030 K/T} + 1.50 x 10(-13) exp{515 K/T} cm3 molecule(-1)(s-1), with a value of (8.5 +/- 0.9) x 10-13 cm3 molecule(-1)(s-1) at 298 K. The steep increase in rate constant in the range 550-800 K, which is reported for the first time, implies that direct abstraction of a methyl-H becomes the dominant pathway at temperatures greater than 550 K. However, the data indicates that up to about 800 K direct methyl-H abstraction remains adversely affected by the long-range H bonding attraction between the approaching OH radical and the carboxyl -C(O)OH functionality. PMID- 17125301 TI - Computed structures of two known Yb@C74 isomers. AB - Six isomers of Yb@C74 are considered, namely one cage with isolated pentagons, three isomers with a pentagon-pentagon junction, two structures with one pentagon pentagon pair, and one heptagon. The computations based on the Gibbs energy point out the endohedral derived from the only C74 cage with the isolated pentagons and from a cage with one pentagon-pentagon junction as the major and minor observed isomer, respectively. Temperatures giving the best agreement with the experimental production ratio are evaluated. PMID- 17125302 TI - Binding energies of first row diatomics in the light of the interacting quantum atoms approach. AB - Binding energies of first row diatomics are revisited within the interacting quantum atoms (IQA) approach. This is a formalism in chemical bonding theory based upon the quantum theory of atoms in molecules. It is characterized by the preservation of the energetic identity of atoms within molecules. Quantum mechanically computed binding energies are recovered in IQA as a sum of small atomic deformation energies and large pairwise interaction terms. We show how this partition responds faithfully to chemical intuition, and how the different evolution of deformations and interactions accounts in a unified manner for the subtle variations of the binding energy of these molecules. PMID- 17125303 TI - Hydride affinities of some substituted alkynes: prediction by DFT calculations and rationalization by triadic formula. AB - Hydride affinities (HAs) of the ethynes substituted by a wide range of different substituents are considered by using the B3LYP methodology. The computed values are in fair agreement with available experimental data, which are unfortunately scarce. The trend of changes of the HAs is rationalized by trichotomy formula. One of the important results of this analysis is a finding that similar HA values might result from completely different effects. Alternative sites of the H- attack are examined and the difference in energies relative to the most susceptible positions is interpreted. Structural features of substituted ethyne hydrides are briefly discussed. PMID- 17125304 TI - Full spectral decomposition of ring currents. AB - Within the ipsocentric method for calculation of molecular magnetic response, projection of perturbed orbitals onto the virtual orbital space allows partition of induced current density into contributions from individual virtual excitations between occupied and unoccupied orbitals, enabling detailed assignment of the origin of currents in, e.g., benzene, cyclooctatetraene, borazine, coronene, and corannulene. Whereas delocalized currents in benzene and planar cyclooctatetraene are described by transitions within the valence space, localized currents in the borazine pi system involve excitations outside the valence space. PMID- 17125305 TI - Study of hydrogen bonding in liquid crystalline solvent by Fourier transform infrared spectroscopy. AB - A hydrogen-bonded complex between an aromatic acid and an enantiopure chiral amine has been dissolved in a nematic solvent, giving rise to a cholesteric medium. Fourier transform infrared (FT-IR) experiments have been performed at various temperatures on both sides of the cholesteric-isotropic transition. Liquid crystalline order provides significant enhancement to the strength of interaction, inducing a discontinuous jump in concentration of the complex at the cholesteric-isotropic transition. PMID- 17125306 TI - Theoretical studies on the ene reaction mechanisms of propene and cyclopropene with ethylene and cyclopropene: concerted or stepwise. AB - The potential energy surfaces of the ene reactions of propene and cyclopropene with ethylene and cyclopropene were studied by ab initio molecular orbital (MO) methods. The reaction mechanisms were analyzed by CiLC method on the basis of CASSCF MOs. The concerted and stepwise reaction pathways of the ene reaction of propene with ethylene as the parent reaction were located. The energy barrier of the stepwise process is about 4 kcal/mol lower than that of the concerted one. The other reactions can be found only the stepwise mechanism. Although the endo type reaction of propene with cyclopropene, where cyclopropene is the enophile, probably occurs through a one-step process, the mechanism is divided into the CC bond formations and the hydrogen migration as a stepwise reaction. The CiLC-IRC analysis of the concerted process of propene with ethylene shows the different patterns of the electronic state variation for the CC bond formation/breaking and the hydrogen migration. PMID- 17125307 TI - Time-dependent density functional theory study of Fe2(CO)9 low-lying electronic excited states. AB - The valence electronic excited states of Fe2(CO)9 have been studied using the time-dependent density functional theory (TDDFT). Both tribridged D3h and monobridged C2v structures have been considered, and the structure of selected low-lying singlet and triplet excited states have been optimized on the basis of the TDDFT analytical gradient. Optimized excited-state geometries are used to obtain an insight into certain aspects of the Fe2(CO)9 photochemistry. The Fe2(CO)9 (D3h) first triplet and second singlet excited states are unbound with respect to dibridged Fe2(CO)8 + CO, and the first two monobridged Fe2(CO)9 (C2v) singlet states are unbound with respect to the Fe(CO)5 + Fe(CO)4 dissociation. These results are discussed in light of the experimental data available. PMID- 17125308 TI - Time dependent density functional theory modeling of specific rotation and optical rotatory dispersion of the aromatic amino acids in solution. AB - Time Dependent Density Functional Theory (TDDFT) along with the COnductor-like Screening MOdel (COSMO) has been applied to model the specific rotation at 589.3 nm and the optical rotatory dispersion (ORD) of the aromatic amino acids phenylalanine, tyrosine, histidine, and tryptophan. Solution structures at low, neutral, and high pH were determined. Both the anomalous dispersion absorbing (resonance) region and the lower energy (transparent) region of the ORD of the compounds were modeled. Linear response calculation of the specific rotation and ORD as well as Kramers-Kronig transformations of calculated circular dichroism spectra to model resonant ORD were compared with experimental data from the literature. PMID- 17125309 TI - A combined QM/MM molecular dynamics simulations study of nitrate anion (NO3-) in aqueous solution. AB - The structural and dynamical properties of NO3- in dilute aqueous solution have been investigated by means of two combined quantum mechanics/molecular mechanics (QM/MM) molecular dynamics simulations, namely HF/MM and B3LYP/MM, in which the ion and its surrounding water molecules were treated at HF and B3LYP levels of accuracy, respectively, using the DZV+ basis set. On the basis of both HF and B3LYP methods, a well-defined first hydration shell of NO3- is obtainable, but the shell is quite flexible and the hydrogen-bond interactions between NO3- and water are rather weak. With respect to the detailed analysis of the geometrical arrangement and vibrations of NO3-, the experimentally observed solvent-induced symmetry breaking of the ion is well reflected. In addition, the dynamical information, i.e., the bond distortions and shifts in the corresponding bending and stretching frequencies as well as the mean residence time of water molecules surrounding the NO3- ion, clearly indicates the "structure-breaking" ability of this ion in aqueous solution. From a methodical point of view it seems that both the HF and B3LYP methods are not too different in describing this hydrated ion by means of a QM/MM simulation. However, the detailed analysis of the dynamics properties indicates a better suitability of the HF method compared to the B3LYP DFT approach. PMID- 17125310 TI - Calculation of electron affinities of polycyclic aromatic hydrocarbons and solvation energies of their radical anion. AB - Electron affinities (EAs) and free energies for electron attachment (DeltaGo(a,298K)) have been directly calculated for 45 polynuclear aromatic hydrocarbons (PAHs) and related molecules by a variety of theoretical methods, with standard regression errors of about 0.07 eV (mean unsigned error = 0.05 eV) at the B3LYP/6-31 + G(d,p) level and larger errors with HF or MP2 methods or using Koopmans' Theorem. Comparison of gas-phase free energies with solution phase reduction potentials provides a measure of solvation energy differences between the radical anion and neutral PAH. A simple Born-charging model approximates the solvation effects on the radical anions, leading to a good correlation with experimental solvation energy differences. This is used to estimate unknown or questionable EAs from reduction potentials. Two independent methods are used to predict DeltaGo(a,298K) values: (1) based upon DFT methods, or (2) based upon reduction potentials and the Born model. They suggest reassignments or a resolution of conflicting experimental EAs for nearly one-half (17 of 38) of the PAH molecules for which experimental EAs have been reported. For the antiaromatic molecules, 1,3,5-tri-tert-butylpentalene and the dithia substituted cyclobutadiene 1, the reduction potentials lead to estimated EAs close to those expected from DFT calculations and provide a basis for the prediction of the EAs and reduction potentials of pentalene and cyclobutadiene. The Born model has been used to relate the electrostatic solvation energies of PAH and hydrocarbon radical anions, and spherical halide anions, alkali metal cations, and ammonium ions to effective ionic radii from DFT electron-density envelopes. The Born model used for PAHs has been successfully extended here to quantitatively explain the solvation energy of the C60 radical anion. PMID- 17125311 TI - Transverse acoustic modes of biogenic and alpha,omega-polyamines: a study by inelastic neutron scattering and Raman spectroscopies coupled to DFT calculations. AB - A complete analysis of the transverse acoustic modes (TAMs) for the homologous series of alpha,omega-diamines (H2N(CH2)nNH2) (n = 2-10, n = 12) as well as for the biogenic polyamines spermidine and spermine was undertaken, by Raman and inelastic neutron scattering (INS) spectroscopies combined with density functional theory (DFT) calculations. A complete assignment of the whole set of TAMs was carried out, for both the undeuterated and N-deuterated species. 1,2 Diaminoethane was found to display exceptional behavior, probably due to the formation of dimers in the solid state. An n-even/n-odd dependence of the low frequency INS pattern was observed for these polyamines. The very good accordance between their INS experimental TAMs and the ones previously reported for the corresponding n-alkanes suggest a close conformational similarity between these systems. PMID- 17125312 TI - Sigma- and pi-bond strengths in main group 3-5 compounds. AB - The sigma- and pi-bond strengths for the molecules BH2NH2, BH2PH2, AlH2NH2, and AlH2PH2 have been calculated by using ab initio molecular electronic structure theory at the CCSD(T)/CBS level. The adiabatic pi-bond energy is defined as the rotation barrier between the equilibrium ground-state configuration and the C(s)symmetry transition state for torsion about the A-X bond. We also report intrinsic pi-bond energies corresponding to the adiabatic rotation barrier corrected for the inversion barrier at N or P. The adiabatic sigma-bond energy is defined as the dissociation energy of AH2XH2 to AH2 + XH2 in their ground states minus the adiabatic pi-bond energy. The adiabatic sigma-bond strengths for the molecules BH2NH2, BH2PH2, AlH2NH2, and AlH2PH2 are 109.8, 98.8, 77.6, and 68.3 kcal/mol, respectively, and the corresponding adiabatic pi-bond strengths are 29.9, 10.5, 9.2, and 2.7 kcal/mol, respectively. PMID- 17125313 TI - Frequency analysis of amide-linked rotaxane mimetics. AB - A vibrational analysis of 2-fold hydrogen bonds between an isophthalic amide donor and different acceptors is presented. These systems can be considered as mimetics for the hydrogen-binding situation of numerous supramolecular compounds such as rotaxanes, catenanes, knotanes, and anion receptors. We calculated pronounced red-shifts up to 65 cm(-1) for the stretching modes of the acceptor carbonyl as well as for the donor NH2 groups, whereas we observe a blue shift for the NH2 bending modes and an additional weak hydrogen bond between the acceptor and the middle C-H group of the donor. The red and blue shifts observed for different modes in various complexes have been correlated with the binding energy of the complexes, independently. In comparison with comparable single hydrogen bonds, we find for the 2-fold hydrogen bonds smaller red shifts for the N-H stretch modes of the donor but larger red shifts for the C=O stretch mode of the acceptor. Furthermore, our results indicate that the pronounced blue shift of the C-H stretch mode is basically caused by the fact that the acceptor is fixed directly above this group due to the 2-fold hydrogen bond. PMID- 17125314 TI - Donor-acceptor-donor tetrazines containing a ferrocene unit: synthesis, electrochemical and spectroscopic properties. AB - Donor-acceptor-donor tetrazines containing ferrocene moieties and phenyl unit as a pi-bridge have been synthesized and characterized. UV-vis spectroscopic and cyclic voltamperometric results indicate sizable intramolecular charge transfer interactions in the ground state when the ferrocene is directly bound to the tetrazine. On the other hand, the results show reduction of the electron-donor strength of ferrocene moieties when there is a phenyl linkage. Both tetrazines display a high reduction potential. The role of ferrocenyl groups appear to be detrimental to maximize the cubic hyperpolarizability gamma of tetrazines, as compared to purely organic groups such as thiophene. A possible explanation for this behavior may originate from metal-to-ligand charge transfer processes. PMID- 17125315 TI - Scanning tunneling microscopy and spectroscopy of wet-chemically prepared chlorinated Si111 surfaces. AB - Chlorine-terminated Si(111) surfaces prepared through the wet-chemical treatment of H-terminated Si(111) surfaces with PCl5 (in chlorobenzene) were investigated using ultrahigh vacuum scanning tunneling microscopy (UHV cryo-STM) and tunneling spectroscopy. STM images, collected at 77 K, revealed an unreconstructed 1 x 1 structure for the chlorination layer, consistent with what has been observed for the gas phase chlorination of H-terminated Si(111). However, the wet-chemical chlorination is shown to generate etch pits in the Si(111) surface, with an increase in etch pit density correlating with increasing PCl5 exposure temperatures. These etch pits were assumed to stabilize the edge structure through the partial removal of the <112> step edges. Tunneling spectroscopy revealed a nonzero density of states at zero bias. This is in contrast to the cases of H-, methyl-, or ethyl-terminated Si(111), in which similar measurements have revealed the presence of a large conductance gap. PMID- 17125316 TI - Structure, bonding nature, and binding energy of alkanethiolate on As-rich GaAs (001) surface: a density functional theory study. AB - Chemisorption of alkanethiols on As-rich GaAs (001) surface under a low coverage condition was studied using first principles density functional calculations in a periodic supercell approach. The thiolate adsorption site, tilt angle and its direction are dictated by the high directionality of As dangling bond and sulfur 3p orbital participating in bonding and steric repulsion of the first three CH2 units from the surface. Small charge transfer between thiolate and surface, strong dependence of total energy on tilt angle, and a relatively short length of 2.28 A of the S-As bond indicate the highly covalent nature of the bonding. Calculated binding energy of 2.1 eV is consistent with the available experimental data. PMID- 17125317 TI - Conformational selectivity of peptides for single-walled carbon nanotubes. AB - Carbon nanotubes show promising prospects for applications ranging from molecular electronics to ultrasensitive biosensors. An important aspect to understanding carbon nanotube properties is their interactions with biomolecules such as peptides and proteins, as these interactions are important in our understanding of nanotube interactions with the environment, their use in cellular systems, as well as their interface with biological materials for medical and diagnostic applications. Here we report the sequence and conformational requirements of peptides for high-affinity binding to single-walled carbon nanotubes (SWNTs). A new motif, X(1)THX(2)X(3)PWTX(4), where X(1) is G or H, X(2) is H or D or null, X(3) is null or R, and X4 is null or K, was identified from two classes of phage displayed peptide libraries. The high affinity binding of the motif to SWNTs required constrained conformations which were achieved through either the extension of the amino acid sequence (e.g., LLADTTHHRPWT) or the addition of a constrained disulfide bond (e.g., CGHPWTKC). This motif shows specific high affinity to the currently studied SWNTs, compared to previously reported peptides. The conformations of the identified peptides in complex with SWNTs were also characterized with a variety of biophysical methodologies including CD, fluorescence, NMR spectroscopy, and molecular modeling. PMID- 17125318 TI - How do aryl groups attach to a graphene sheet? AB - How aryl groups attach to a graphene sheet is an experimentally unanswered question. Using first principles density functional theory methods, we shed light on this problem. For the basal plane, isolated phenyl groups are predicted to be weakly bonded to the graphene sheet, even though a new single C-C bond is formed between the phenyl group and the basal plane by converting a sp2-carbon in the graphene sheet to sp3. However, the interaction can be strengthened significantly with two phenyl groups attached to the para positions of the same six-membered ring to form a pair on the basal plane. The strongest bonding is found at the graphene edges. A 1,2-addition pair is predicted to be most stable for the armchair edge, whereas the zigzag edge possesses a unique localized state near the Fermi level that shows a high affinity for the phenyl group. PMID- 17125319 TI - Density functional theory study of geometrical structures and electronic properties of silica nanowires. AB - Silica nanowires are expected to possess structural diversity like bulk silica. We modeled three silica nanowires based on the side-shared two-membered rings, spiro-united two-membered rings, and three-membered rings, respectively. By performing density functional theory calculations, we studied their geometrical structures and electronic properties with and without the presence of external electric field. It is found that the stability of silica nanowires increases with length and diameter. As indicated by calculated large HOMO-LUMO gaps, silica nanowires are expected to be good insulating materials. The energy gaps, however, gradually decrease with applied electronic field and finally close, resulting in the breakdown of the insulating nanowires. Moreover, it is shown that the breakdown threshold remarkably increases with the nanowire diameter. These significant findings from the present calculations for the simplest silica nanowires will provide relevant insight into the structures and properties of much more complicated real silica nanowires. PMID- 17125320 TI - Structural and magnetic properties of Gd3N@C80. AB - Using relativistic and on-site correlation-corrected density functional theory, we have investigated the structural and magnetic properties of recently synthesized Gd3N@C80. The most stable structure of Gd3N@C80 has the three magnetic Gd ions pointing to the centers of hexagons in C80. The magnetic ground state of this structure has the three coplanar spins (S = 7/2) offset by 120 degrees angles. At the same time, the state with the highest multiplicity, where all the spins are parallel aligned, is found only about 4.5 meV higher in energy. Therefore, at room temperature, we expect Gd3N@C80 to be paramagnetic with the spin fluctuating between different multiplicities. As a result, Gd3N@C80 may exhibit greater proton relaxivity than Gd@C60 and Gd@C82 and serve as a possible candidate for the next generation of commercially available magnetic resonance imaging contrast agents. PMID- 17125321 TI - Quantum dynamics of the H + O2 --> O + OH reaction on an accurate ab initio potential energy surface. AB - We report exact time-dependent and time-independent quantum mechanical studies of the title reaction on an accurate ab initio potential energy surface of Xu et al. (J. Chem. Phys. 2005, 122, 24305). The J = 0 reaction probabilities for several reactant states show sharp resonance structures superimposed on relatively low backgrounds, and they are remarkably different from existing quantum results on an earlier potential energy surface (DMBE-IV). The new findings reported here suggest that our current understanding of this important reaction might require significant revision. PMID- 17125322 TI - Molecular simulation analysis and X-ray absorption measurement of Ca2+, K+ and Cl ions in solution. AB - This paper presents recent advances in the use of molecular simulations and extended X-ray absorption fine structure (EXAFS) spectroscopy, which enable us to understand solvated ions in solution. We report and discuss the EXAFS spectra and related properties governing solvation processes of different ions in water and methanol. Molecular dynamics (MD) trajectories are coupled to electron scattering simulations to generate the MD-EXAFS spectra, which are found to be in very good agreement with the corresponding experimental measurements. From these simulated spectra, the ion-oxygen distances for the first hydration shell are in agreement with experiment within 0.05-0.1 A. The ionic species studied range from monovalent to divalent, positive and negative: K+, Ca2+, and Cl-. This work demonstrates that the combination of MD-EXAFS and the corresponding experimental measurement provides a powerful tool in the analysis of the solvation structure of aqueous ionic solutions. We also investigate the value of electronic structure analysis of small aqueous clusters as a benchmark to the empirical potentials. In a novel computational approach, we determine the Debye-Waller factors for Ca2+, K+, and Cl- in water by combining the harmonic analysis of data obtained from electronic structure calculations on finite ion-water clusters, providing excellent agreement with the experimental values, and discuss how they compare with results from a harmonic classical statistical mechanical analysis of an empirical potential. PMID- 17125323 TI - Crystal structure of the high-pressure phase of hexahydro-1,3,5-trinitro-1,3,5 triazine (gamma-RDX). AB - The crystal structure of the high-pressure phase of hexahydro-1,3,5-trinitro 1,3,5-triazine (gamma-RDX), which is stable above 4 GPa at room temperature, was investigated by using infrared spectroscopy and powder X-ray diffraction measurements followed by Rietveld refinements using a diamond anvil cell (DAC). Although gamma and alpha phases were found to belong to the same space group Pbca, they exhibited a different crystal packing. The molecular structure of the gamma phase exhibited the same conformation as that of the alpha phase; however, the torsion angles of N-NO2 changed marginally. PMID- 17125324 TI - Electron-beam-induced topographical, chemical, and structural patterning of amorphous titanium oxide films. AB - Electrolytically deposited amorphous TiO2 films on steel are remarkably sensitive to electron beam (e-beam) irradiation at moderate energies at 20 keV, resulting in controlled local oxide reduction and crystallization, opening the possibility for local topographical, chemical, and structural modifications within a biocompatible, amorphous, and semiconducting matrix. The sensitivity is shown to vary significantly with the annealing temperature of as-deposited films. Well defined irradiation conditions in terms of probe current IP (5 microA) and beam size were achieved with an electron probe microanalyzer. As shown by atomic force and optical microscopy, micro-Raman spectroscopy, wavelength-dispersive X-ray (WDX), and Auger analyses, e-beam exposure below 1 Acm-2 immediately leads to electron-stimulated oxygen desorption, resulting in a well-defined volume loss primarily limited to the irradiated zone under the electron probe and in a blue color shift in this zone because of the presence of Ti2O3. Irradiation at 5 Acm( 2) (IP = 5 microA) results in local crystallization into anatase phase within 1 s of exposure and in reduction to TiO after an extended exposure of 60 s. Further reduction to the metallic state could be observed after 60 s of exposure at approximately 160 Acm(-2). The local reduction could be qualitatively sensed with WDX analysis and Auger line scans. An estimation of the film temperature in the beam center indicates that crystallization occurs at less than 150 degrees C, well below the atmospheric crystallization temperature of the present films. The high e-beam sensitivity in combination with the well-defined volume loss from oxygen desorption allows for precise electron lithographic topographical patterning of the present oxides. Irradiation effects leading to the observed reduction and crystallization phenomena under moderate electron energies are discussed. PMID- 17125325 TI - External electric field effects on fluorescence of pyrene butyric acid in a polymer film: concentration dependence and temperature dependence. AB - Fluorescence spectra and electrofluorescence spectra (plots of the electric field induced change in fluorescence intensity as a function of wavelength) have been measured at different temperatures for pyrene butyric acid (PBA) in a PMMA film at different concentrations. At a low concentration of 0.5 mol % where fluorescence emitted from the locally excited state of PBA (LE fluorescence) is dominant, LE fluorescence spectra show only the Stark shift in the presence of an electric field (F), which results from the difference in molecular polarizability between the ground and emitting states. At a high concentration of 10 mol % where the so-called sandwich-type excimer fluorescence (EX(1)) is dominant, both EX(1) and LE fluorescence are quenched by F. Another fluorescence assigned to a partially overlapped excimer (EX(2)) also exists at room temperature, and this emission is enhanced by F. As the temperature decreases, three fluorescence emissions whose electric field effects are different from each other become clear besides EX(1) and LE fluorescence, indicating that at least five fluorescence components exist at high concentrations at low temperatures. At a medium concentration of 5 mol % where EX(1) is comparable in intensity to the LE fluorescence, the intensity of EX(1) is not affected by F at any temperature, but LE fluorescence and EX(2) are markedly influenced by F at room temperature, and four fluorescence emissions are confirmed at low temperatures. PMID- 17125326 TI - Enhanced diffraction efficiency in a photorefractive liquid crystal cell with poly(9-vinylcarbazole)-infiltrated mesoporous TiO2 layers. AB - The photorefractive effect of a layer-structured liquid crystal cell was significantly enhanced when a C60-doped poly(9-vinylcarbazole) (PVK)/TiO2 nanocomposite was used in two photoconductive layers. The C60-doped PVK/TiO2 nanocomposite film was prepared by infiltrating C60-doped PVK into a highly ordered mesoporous TiO2 layer. The addition of the TiO2 layer to the C60-doped PVK layer increased the first-order Raman-Nath diffraction efficiency from 24% to 42.9%. This enhancement of diffraction efficiency is attributed to a blocking effect of charge recombination in the composite layer. The electron transfer from the PVK layer into the TiO2 layer would decrease the recombination of photogenerated charges in the PVK layer, while charges in the PVK layer could participate in the formation of a space-charge field. PMID- 17125327 TI - EPR studies on molecular orientation in a surface-stabilized paramagnetic liquid crystal cell. AB - By EPR spectroscopy, we have developed a new method for determining the molecular orientation in a surface-stabilized liquid crystal (LC) cell, which includes a paramagnetic LC, (2S,5S)-2,5-dimethyl-2-heptyloxyphenyl-5-[4-(4 octyloxybenzenecarbonyloxy)phenyl]pyrrolidine-1-oxy (1), whose spin source is fixed in the rigid core. For each phase of racemic [(+/-)] and enantiomerically enriched [(S,S)] 1 in a surface-stabilized LC cell (4 microm thickness), the observed g-value profiles depending on the angle between the applied magnetic field and the cell plane were successfully simulated by the orientation models: (i) the LC molecule in the nematic (N) phase of (+/-)-1 freely rotates around the long axis, which is always parallel to the rubbing direction; (ii) the long axis of the freely rotating LC molecule in the chiral nematic (N*) phase of (S,S)-1 is always parallel to the cell plane but rotates in the plane to form a helical superstructure; and (iii) in the crystalline phase of (S,S)-1, the molecular long axis forms a helical superstructure similar to that of the N* phase, but the molecule is fixed around the long axis so that the NO bond lies in the cell plane. Fitting the temperature profile of the g-value in the N phase of (+/-)-1 by use of the Haller equation, we determined the molecular g-values along the molecular long axis (g(parallelM)) and short axis (g(perpendicularM)), which were successfully reproduced by the use of the set of principal g-values of a similar nitroxide with consideration of the structure of the LC molecule optimized by Molecular Mechanics 3 (MM3). PMID- 17125328 TI - Influence of the precipitation agent in the deposition-precipitation on the formation and properties of Au nanoparticles supported on Al2O3. AB - Au nanoparticles supported on Al2O3 were prepared by deposition-precipitation of HAuCl4 with different precipitation agents NaOH and urea. The samples were investigated by means of different characterization techniques such as X-ray photoelectron spectroscopy (XPS), X-ray absorption spectroscopy (XAS), and transmission electron microscopy (TEM). The results show that depending on the precipitation agent, the Au particles have a different Au-Au coordination number and size after calcination at 523 K. Whereas the use of NaOH leads to the formation of Au nanoparticles with a Au-Au coordination number of 6.7 and a mean diameter below 2 nm, those prepared with urea have a mean size of 3.1 nm. The Au Au coordination number could be determined as 8.6. At the smaller particles obtained with NaOH, hints for Au-O interactions were found. For these particles TEM results advise a rather flat lenticular morphology. Different deposition mechanisms depending on the precipitation agent are discussed as the reason for the formation of nanoparticles with different shapes, sizes, and valence states. PMID- 17125329 TI - Growth and structure of carbon nanotube Y-junctions. AB - The effect of a catalyst on the growth and structure of carbon nanotube Y junctions (CNTYs) using chemical vapor deposition has been investigated. Cobalt-, magnesium-, and calcium-nitrates are utilized as precursors of catalysts Co, Mg, Ca, Co/Mg, Co/Ca, and Mg/Ca for CNTY synthesis. Experimental result shows that Co/Mg or Co/Ca can grow CNTYs with straight branches while Co, Mg, Ca, and Mg/Ca will not grow any CNTYs, indicating that only combinations of Co with Mg or Ca will facilitate the formation of CNTYs. In addition, the effect of the carbon source on the formation of CNTYs has also been studied. It is found that thiophene (C4H4S) can promote the formation of CNTYs, while other sources such as methane (CH4) and acetylene (C2H2) cannot. The result shows that both the catalyst and the carbon source substantially affect the formation of CNTYs. PMID- 17125330 TI - Keto forms of salicylaldehyde Schiff bases: structural and theoretical aspects. AB - Twelve Schiff bases of methoxy-substituted salicylaldehyde have been examined by crystallographic and spectroscopic methods, as well as by DFT theoretical calculations in order to investigate the effect of the substituent's position on the keto-enol equilibrium in the crystalline state. Four out of the 10 structurally characterized compounds with methoxy substitution on the para and/or ortho positions with respect to the aldimine bridge and deriving from aliphatic amines or alkylarylamines are found as cis-keto tautomers and form dimers. In contrast, the five pure enol tautomers derive either from aliphatic or alkylarylamines and are meta substituted or from aniline or benzylamine and are para and/or ortho methoxy substituted. The DFT calculations support the crystallographic results and, moreover, they have shown that keto and enol tautomers are affected differently by the relative arrangement of the monomers. Overall, the DFT calculations point to a plausible hypothesis for the stabilization of the keto form in the crystalline state: In cases with a sufficiently low enol-keto energy difference of the isolated monomers, as when the methoxy group is at ortho and/or para positions with respect to the aldimino group, extra stabilization of the keto form is derived from molecular association, thus leading to its crystallization. PMID- 17125331 TI - Loading behavior of {chitosan/hyaluronic acid}n layer-by-layer assembly films toward myoglobin: an electrochemical study. AB - When {CS/HA}n layer-by-layer films assembled by oppositely charged chitosan (CS) and hyaluronic acid (HA) were immersed in myoglobin (Mb) solution at pH 5.0, Mb was gradually loaded into the {CS/HA}n films, designated as {CS/HA}n-Mb. The cyclic voltammetric (CV) peak pair of Mb FeIII/FeII redox couple for {CS/HA}n-Mb films on pyrolytic graphite (PG) electrodes was used to investigate the loading behavior of {CS/HA}n films toward Mb. The various influencing factors, such as the number of bilayers (n), the pH of Mb loading solution, and the ionic strength of solution, were investigated by different electrochemical methods and other techniques. The results showed that the main driving force for the bulk loading of Mb was most probably the electrostatic interaction between oppositely charged Mb in solution and HA in the films, while other interactions such as hydrogen bonding and hydrophobic interaction may also play an important role. Other polyelectrolyte multilayer (PEM) films with different components were compared with {CS/HA}n films in permeability and Mb loading, and electroactive probes with different size and surface charge were compared in their incorporation into PEM films. The results suggest that due to the unique structure of CS and HA, {CS/HA}n films with relatively low charge density are packed more loosely and more easily swelled by water, and have better permeability, which may lead to the higher loading amount and shorter loading time for Mb. The protein-loaded PEM films provide a new route to immobilize redox proteins on electrodes and realize the direct electrochemistry of the proteins. PMID- 17125332 TI - In situ and frontal polymerization for the consolidation of porous stones: a unilateral NMR and magnetic resonance imaging study. AB - Consolidation treatment of porous materials was performed by in situ and frontal polymerization of acrylic monomers inside a porous stone. To study the penetration of the polymer inside the stone and its consolidating effects we used water as a contrast agent, detecting its penetration using unilateral NMR and magnetic resonance imaging. All data obtained on differently treated stones were compared with corresponding ones obtained analyzing both untreated stones and stones simply painted with a well-known polymeric protective agent. In situ polymerization of acrylic monomers inside porous stones has been demonstrated to be an extremely powerful consolidating method, whereas thermally initiated frontal polymerization seems less efficient. In both cases the optimal choice of monomers is still open and requires further study. Our data indicate that unilateral NMR represents an inexpensive and simple technique for the non invasive observation of the water uptake and of the effect of consolidation procedures in porous materials. PMID- 17125333 TI - Ordered structures in proton conducting membranes from supramolecular liquid crystal polymers. AB - Highly sulfonated forms of poly(p-phenylene terephthalamide) (PPTA) have been prepared in three different molecular configurations; sulfonated diamine form (S PPTA), sulfonated terephthalic acid form (S-invert-PPTA), and the bi-sulfonated form (S2-PPTA). All three polymers are water soluble to a certain degree and films were cast from solution for S-PPTA and S-invert-PPTA. S-PPTA films absorb less water than S-invert-PPTA (under controlled humidity conditions) and consequently, the conductivity for this polymer is also slightly lower. Although the conductivities are comparable to Nafion (of the order of 10(-2) to 10(-1) Scm(-1)), proton mobility is more restricted. X-ray diffraction showed that the rigid molecules are aligned in opposite directions for the two polymer films, being homeotropic in S-PPTA films and planar for S-invert-PPTA. SEM analysis demonstrated layering in the same direction as the alignment of the polymer chains. The variation in the polymer alignment is most likely the result of the differences in the solution properties and the film forming process. It is possible, however, that this alignment could be exploited to enhance proton transport and thus these films are of interest for fuel cell membranes. PMID- 17125334 TI - pH-dependent electron-transport properties of carbon nanotubes. AB - Carbon nanotube electrochemical transistors integrated with microfluidic channels are utilized to examine the effects of aqueous electrolyte solutions on the electron-transport properties of single isolated carbon nanotubes. In particular, pH and concentration of supporting inert electrolytes are examined. A systematic threshold voltage shift with pH is observed while the transconductance and subthreshold swing remain independent of pH and concentration. Decreasing pH leads to a negative shift of the threshold voltage, indicating that protonation does not lead to hole doping. Changing the type of contact metal does not alter the observed pH response. The pH-dependent charging of SiO2 substrate is ruled out as the origin based on measurements with suspended nanotube transistors. Increasing the ionic strength leads to reduced pH response. Contributions from possible surface chargeable chemical groups are considered. PMID- 17125335 TI - Investigation of field emission and photoemission properties of high-purity single-walled carbon nanotubes synthesized by hydrogen arc discharge. AB - Single-walled carbon nanotubes (SWCNTs) were directly synthesized by a hydrogen arc-discharge method by using only Fe catalyst. The synthesized carbon materials indicated high-purity SWCNTs without amorphous carbon materials from SEM observation. The SWCNTs had diameters of 1.5-2.0 nm from TEM and Raman observation. After a simple purification, TGA indicated that SWCNTs had a purity of ca. 90.1 wt %. Field emission from the SWCNT emitters which were fabricated by using a spray method was measured by a diode structure. The vertically aligned SWCNT emitters showed the low turn-on voltage of 0.86 V/microm and a high emission-current density of 3 mA/cm2 at an applied field of about 3 V/microm. From a Fowler-Nordheim plot, the vertically aligned SWCNT revealed a high field enhancement factor of 2.35 x 10(4). The photoemission measurements, excited by a photon energy of 360 eV, showed significantly delocalized graphite-pi states at the purified SWCNTs. Here, we investigated that the field-emission properties of SWCNTs would be attributed to the high electronic density of states near Fermi energy, including the delocalized graphite-pi states. PMID- 17125336 TI - Theoretical investigation of the tunable behavior of p-n copolymers based on oligothiophenes and 1,4-bis(oxadiazolyl)-benzene. AB - Semiempirical calculations were carried out on several model oligomers to investigate the tunable behavior of p-n copolymers with the repeating units constructed by oligothiophenes as the pi-excessive type blocks and 1,4 bis(oxadiazolyl) benzene as the pi-deficient type block. The calculated evolutions of the HOMO and LUMO of the model oligomers were in good agreement with the experimental oxidation and reduction potentials of the corresponding polymers. The effect of the length of the oligothiophene on the electronic structures and optical properties was elucidated by analyzing the HOMO and LUMO spatial distribution patterns of the model oligomers. When the number of thiophene rings increases, the HOMO and LUMO are contributed mostly from the oligothiophene segments and either the introduced single positive or negative charge focuses on the oligothiophene segments. The absorption spectra of polymers were also simulated by performing calculations on the corresponding oligomers. Good matches were observed between the calculated absorption spectra and the experimental UV-vis spectra of the corresponding polymers. The study shows that the backbone modification of the p-n copolymer, that is, changing the number of thiophene unit in the p-n diblock copolymer, greatly modifies the optical properties of the polymer. PMID- 17125338 TI - Optimization of slitlike carbon nanopores for storage of hythane fuel at ambient temperatures. AB - Carbons with slitlike pores can serve as effective host materials for storage of hythane fuel, a bridge between the petrol combustion and hydrogen fuel cells. We have used grand canonical Monte Carlo simulation for the modeling of the hydrogen and methane mixture storage at 293 K and pressure of methane and hydrogen mixture up to 2 MPa. We have found that these pores serve as efficient vessels for the storage of hythane fuel near ambient temperatures and low pressures. We find that, for carbons having optimized slitlike pores of size H congruent with 7 A (pore width that can accommodate one adsorbed methane layer), and bulk hydrogen mole fraction >or=0.9, the volumetric stored energy exceeds the 2010 target of 5.4 MJ dm(-3) established by the U.S. FreedomCAR Partnership. At the same condition, the content of hydrogen in slitlike carbon pores is approximately = 7% by energy. Thus, we have obtained the composition corresponding to hythane fuel in carbon nanospaces with greatly enhanced volumetric energy in comparison to the traditional compression method. We proposed the simple system with added extra container filled with pure free/adsorbed methane for adjusting the composition of the desorbed mixture as needed during delivery. Our simulation results indicate that light slit pore carbon nanomaterials with optimized parameters are suitable filling vessels for storage of hythane fuel. The proposed simple system consisting of main vessel with physisorbed hythane fuel, and an extra container filled with pure free/adsorbed methane will be particularly suitable for combustion of hythane fuel in buses and passenger cars near ambient temperatures and low pressures. PMID- 17125337 TI - Structure and bonding of the multifunctional amino acid L-DOPA on Au(110). AB - In investigations of the proteins which are responsible for the surface adhesion of the blue mussel Mytilus edulis, an unusually frequent appearance of the otherwise rare amino acid 3-(3,4-dihydroxyphenyl)-L-alanine (L-DOPA) has been observed. This amino acid is thought to play a major role in the mechanism of mussel adhesion. Here we report a detailed structural and spectroscopic investigation of the interface between L-DOPA and a single-crystalline Au(110) model surface, with the aim of understanding fundamentals about the surface bonding of this amino acid and its role in mussel adhesion. Molecular layers are deposited by organic molecular beam deposition (OMBD) in an ultrahigh-vacuum environment. The following experimental techniques have been applied: ex situ Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), low-energy electron diffraction (LEED), high-resolution electron energy loss spectroscopy (HREELS), and scanning tunneling microscopy (STM). Vibrational spectra of isolated L-DOPA molecules and the zwitterionic bulk have been calculated using density functional theory (DFT). The predicted modes are assigned to observed spectra, allowing conclusions regarding the molecule substrate and molecule-molecule interactions at the L-DOPA/Au(110) interface. We find that zwitterionic L-DOPA forms a monochiral, one-domain commensurate monolayer on Au(110), with the catechol rings on top of [110] gold rows, oriented parallel to the surface. The (2 x 1)-Au(110) surface reconstruction is not lifted. The carboxylate group is found in a bidentate or bridging configuration, the amino group is tilted out of the surface plane, and the hydroxyl groups do not dehydrogenate on Au(110). Similar to the case for the bulk, molecules form dimers on Au(110). However, the number of hydrogen bridge bonds between L-DOPA molecules is reduced as compared to the bulk. Thicker layers which are deposited onto the commensurate interface do not order in the bulk structure. In conclusion, our study shows that the aromatic ring system of L-DOPA functions as a surface anchor. Since it is also known that the hydroxyl groups support cross link reactions between L-DOPA residues in the mussel glue protein, we can conclude that the catechol ring supports surface adhesion of mussel proteins via two independent functions. PMID- 17125339 TI - Fabrication and characterization of polycrystalline WO3 nanofibers and their application for ammonia sensing. AB - We describe the fabrication and characterization of tungsten oxide nanofibers using the electrospinning technique and sol-gel chemistry. Tungsten isopropoxide sol-gel precursor was incorporated into poly(vinyl acetate)(PVAc)/DMF solutions and electrospun to form composite nanofibers. The as-spun composite nanofibers were subsequently calcinated to obtain pure tungsten oxide nanofibers with controllable diameters of around 100 nm. SEM and TEM were utilized to investigate the structure and morphology of tungsten oxide nanofibers before and after calcination. The relationship between solution concentration and ceramic nanofiber morphology has been studied. A synchrotron-based in situ XRD method was employed to study the dynamic structure evolution of the tungsten oxide nanofibers during the calcination process. It has been shown that the as-prepared tungsten oxide ceramic nanofibers have a quick response to ammonia with various concentrations, suggesting potential applications of the electrospun tungsten oxide nanofibers as a sensor material for gas detection. PMID- 17125340 TI - PET suppression of acridinedione dyes by urea derivatives in water and methanol. AB - Spectroscopic investigations involving the interaction of acridinedione dyes with urea and its derivatives in water and methanol were carried out by absorption, steady-state fluorescence, and time-resolved fluorescence measurements. The hydrogen-bonding properties of urea and derivatives in aqueous solutions are found to be distinctly different from those observed in methanol. Urea, which can serve both as a hydrogen bond donor as well as an acceptor and has a unique hydrogen-bonding feature, helps in studying urea interaction with fluorophores in aqueous solutions, micelles, and alcohol. In our studies, we have used acridinedione dyes as the probe. We report that the hydrophobic interaction of urea with dye predominates by weakening of the hydrogen-bonding interaction of the solvent and urea derivatives with increase in the hydrophobicity of urea derivatives. In methanol, the hydrogen bonding between solvent and urea derivatives predominating over the hydrophobicity of the urea derivatives is observed. The presence of alkyl group substitution in the N-H moiety with a function of increasing concentration resulting in the creation of a more favorable hydrophobic environment to the dye molecule to reside in the hydrophobic shell phase rather than in the bulk aqueous phase is illustrated. The hydrophobic interaction of dye with urea in aqueous solution predominates because of the weakening of the hydrogen bonding of the solvent and urea derivatives, and the photoinduced electron transfer (PET) process is used as a marker to identify the hydrophobic interaction illustrated in our studies. PMID- 17125341 TI - Crystallization and shape evolution of single crystalline selenium nanorods at liquid-liquid interface: from monodisperse amorphous Se nanospheres toward Se nanorods. AB - The reduction of selenious acid solution with hydrazine hydrate in the presence of poly(vinylpyrrolidone) (PVP) can produce a stable dispersion of uniform and amorphous selenium particles capped with PVP with a size of 100 nm. Further addition of a solvent with low polarity such as n-butyl alcohol into this aqueous solution and mild stirring result in the transportation of amorphous selenium particles onto a liquid-liquid interface between water and n-butyl alcohol. Subsequent crystallization and shape evolution on this interface occurred and finally resulted in the formation of single crystalline selenium nanorods. The results demonstrated that the enrichment of nanoparticles with amphiphilic property at a liquid-liquid interface between a polar solvent and another solvent of low polarity can result in crystallization and phase transformation for the formation of nanostructures. PMID- 17125342 TI - Water effects on electron transfer in azurin dimers. AB - Recent experimental and theoretical analyses indicate that water molecules between or near redox partners can significantly affect their electron-transfer (ET) properties. Here, we study the effects of intervening water molecules on the electron self-exchange reaction of azurin (Az) by using a newly developed ab initio method to calculate transfer integrals between molecular sites. We show that the insertion of water molecules in the gap between the copper active sites of Az dimers slows down the exponential decay of the ET rates with the copper-to copper distance. Depending on the distance between the redox sites, water can enhance or suppress the electron-transfer kinetics. We show that this behavior can be ascribed to the simultaneous action of two competing effects: the electrostatic interaction of water with the protein subsystem and its ability to mediate ET coupling pathways. PMID- 17125343 TI - Comparative measures of single-wall carbon nanotube dispersion. AB - Model composites of DNA-wrapped single-wall carbon nanotubes in poly(acrylic acid) are used to evaluate metrics of nanotube dispersion. By varying the pH of the precursor solutions, we introduce a controlled deviation from ideal behavior. On the basis of small-angle neutron scattering, changes in near-infrared fluorescence intensity are strongly correlated with dispersion, while optical absorption spectroscopy and resonant Raman scattering are less definitive. Our results represent the first systematic comparison of currently accepted measures of nanotube dispersion. PMID- 17125344 TI - Designing cyclophane-based molecular wire sensors. AB - We propose a model molecular wire sensor that can detect zerovalent chromium by a strong increase in conductance when included in a metal-molecule-metal junction. An essential part of the sensor is a paracyclophane unit that binds to the metal atom. The nature and the energies of the molecular orbitals change drastically after the complex is formed, resulting in a 10- to 12-fold increase in conductance. PMID- 17125345 TI - Metal filament growth in electrically conductive polymers for nonvolatile memory application. AB - Solution processable polymers that can reproducibly form metal filament by applying voltage are investigated for nonvolatile memory application. Up to present, the understanding of materials enabling to make the metal filament has not been well-documented and the vacuum deposition methods were dominantly used in device fabrication. After screening various polymers, we found that only the polymers having two functionalities, the presence of strongly coordinating heteroatom (S or N) with metal ions and the electrical conductivity, showed the reproducible filament formation behavior. Among the polymers screened, the regiorandom poly(3-hexylthiophene) showed the best switching endurance over 30,000 write-read-erase-read cycles without any switching failure. PMID- 17125346 TI - Large low-field magnetoresistance in nanocrystalline magnetite prepared by sol gel method. AB - Nanocrystalline magnetite Fe3O4 samples with a grain size of about 40 nm have been synthesized by an optimized sol-gel method. The single phase of spinel magnetite was confirmed by both X-ray diffraction and transmission electron microscopy. It has been found that the magnetoresistance of the samples at low field (LFMR) is relatively large, and with the decrease of temperature its value at a field of 0.5 T changes dramatically from -2.5% at 300 K to -17.0% at 55 K. With the further decrease of temperature a sharp drop occurs for the magnitude of the magnetoresistance (MR), regarded as a spin (cluster) glass transition in the surface region of the grains that can be confirmed by the zero-field-cooled and field-cooled magnetization and ac susceptibility measurement. The mechanism of the magnetic and transport properties was discussed. PMID- 17125347 TI - Internal concentration gradients of guest molecules in nanoporous host materials: measurement and microscopic analysis. AB - Evolution of internal concentration profiles of methanol in 2-D pore structure of ferrierite crystal was measured in the pressure range of 0 to 80 mbar with the help of the recently developed interference microscopy technique. The measured profiles showed that both a surface barrier and internal diffusion controlled the kinetics of adsorption/desorption. Furthermore, they indicated that in the main part of the crystal, the z-directional 10-ring channels were not accessible to methanol and that the transport of methanol mainly occurred via 8-ring y directional channels. The roof-like part of the crystal was almost instantaneously filled/emptied during adsorption/desorption, indicating accessible 10-ring channels in this section. The measured profiles were analyzed microscopically with the direct application of Fick's second law, and the transport diffusivity of methanol in ferrierite was determined as a function of adsorbed phase concentration. The transport diffusivity varied by more than 2 orders of magnitude over the investigated pressure range. Transport diffusivities, calculated from measured profiles from small and large pressure step changes, were all found to be consistent. Simulated concentration profiles obtained from the solution of Fick's second law with the calculated functional dependence of diffusivities on concentration compared very well with the measured concentration profiles, indicating validity and consistency of the measured data and the calculated diffusivities. The results indicate the importance of measuring the evolution of concentration profiles as this information is vital in determining (1) the direction of internal transport, (2) the presence of internal structural defects, and (3) surface/internal transport barriers. Such detailed information is available neither from common macroscopic methods since, they measure changes in macroscopic properties and use model assumptions to predict the concentration profiles inside, nor from microscopic methods, since they only provide information on average displacement of diffusing molecules. PMID- 17125348 TI - Controllable assembly of WO3 nanorods/nanowires into hierarchical nanostructures. AB - The controlled synthesis of two novel h-WO3 hierarchical structures made of nanorods/nanowires has been successfully realized in a large scale via a simple hydrothermal method. It is demonstrated that the morphology of the final products is significantly influenced by adding different sulfates. The urchinlike and ribbonlike structures of WO3 can be selectively prepared by adding Rb2SO4 and K2SO4, respectively. The morphology evolvement and the growth mechanism were studied carefully. The sulfate-induced oriented attachment growth mechanism has been proposed for the possible formation mechanism of the ribbonlike sample. For urchinlike products, two growing stages are believed to be involved in the growth process. The current understanding of the growth mechanism of these nanostructures may be potentially applied for designing other oriented or hierarchical nanostructures based on 1D nanoscale building blocks through the direct solution-growth. PMID- 17125349 TI - Interaction of CO with palladium supported on oxidized tungsten. AB - A model catalyst system, palladium on tungsten oxide, has been examined by temperature-programmed desorption and photoemission spectroscopy. The samples were prepared by evaporation of palladium onto an oxidized tungsten foil under ultrahigh vacuum conditions. Mostly three-dimensional (3-D) palladium (Pd) clusters were found to be present on oxidized tungsten (WOx) surfaces at room temperature. Upon annealing to 670 K, the palladium clusters are redispersed and decorated by the WOx surface layer. The nature of the WOx phase on top of the palladium clusters is dependent on the mode of oxidation of the tungsten foil prior to palladium deposition. Mainly W(2+) species decorate palladium deposits on tungsten oxidized at room temperature, while mainly W(4+) species are on top of palladium deposits on the surface oxidized at 1300 K. The appearance of a Pd(n+)-O-W(4+) mixed oxide phase with n < 2 was observed on the oxidized tungsten surface. The substantial reduction (relative to nonannealed samples) of molecular CO coverage induced by annealing is discussed in terms of the changes in chemical composition and morphology of the outermost surface. PMID- 17125350 TI - How small polar molecules protect membrane systems against osmotic stress: the urea-water-phospholipid system. AB - We investigate how a small polar molecule, urea, can act to protect a phospholipid bilayer system against osmotic stress. Osmotic stress can be caused by a dry environment, by freezing, or by exposure to aqueous systems with high osmotic pressure due to solutes like in saline water. A large number of organisms regularly experience osmotic stress, and it is a common response to produce small polar molecules intracellularly. We have selected a ternary system of urea-water dimyristoyl phosphatidylcholine (DMPC) as a model to investigate the molecular mechanism behind this protective effect, in this case, of urea, and we put special emphasis on the applications of urea in skin care products. Using differential scanning calorimetry, X-ray diffraction, and sorption microbalance measurements, we studied the phase behavior of lipid systems exposed to an excess of solvent of varying compositions, as well as lipid systems exposed to water at reduced relative humidities. From this, we have arrived at a rather detailed thermodynamic characterization. The basic findings are as follows: (i) In excess solvent, the thermally induced lipid phase transitions are only marginally dependent on the urea content, with the exception being that the P(beta) phase is not observed in the presence of urea. (ii) For lipid systems with limited access to solvent, the phase behavior is basically determined by the amount (volume) of solvent irrespective of the urea content. (iii) The presence of urea has the effect of retaining the liquid crystalline phase at relative humidities down to 64% (at 27 degrees C), whereas, in the absence of urea, the transition to the gel phase occurs already at a relative humidity of 94%. This demonstrates the protective effect of urea against osmotic stress. (iv) In skin care products, urea is referred to as a moisturizer, which we find slightly misleading as it replaces the water while keeping the physical properties unaltered. (v) In other systems, urea is known to weaken the hydrophobic interactions, while for the lipid system we find few signs of this loosening of the strong segregation into polar and apolar regions on addition of urea. PMID- 17125351 TI - Adsorption and mixing behavior of ethers and alkanes at the solid/liquid interface. AB - The behavior of binary mixtures of linear symmetrical ethers and alkanes adsorbed to a graphite surface from the bulk liquid mixtures is described on the basis of differential scanning calorimetry (DSC) data. Both the ethers and the alkanes are found to form solid monolayers when adsorbed from the liquid. In addition, the monolayer mixing behavior is addressed. The results indicate that there is good, essentially ideal, mixing in the monolayers for ethers and alkanes of the same overall chain length, where the chain length is equal to the total number of carbon and oxygen atoms in the molecule. However, a difference in chain length of more than one atom results in a variation of mixing behavior from nonideal mixing (for long pairs) to phase separation (for short pairs) on the graphite surface. Hence, we conclude that it is the relative chain lengths that control mixing behavior. The results are quantified using a regular solution model with a correction for preferential adsorption. The phase behavior of the mixed monolayers is also compared to the behavior of the bulk. Interestingly, we observe mixtures where the bulk and monolayer behavior are quite different, for example, phase separation in the bulk but essentially ideal mixing in the monolayer for mixtures of ethers and alkanes with the same chain lengths. At present, we attribute this mixing in the monolayer to dilution of the unfavorable ether oxygen-ether oxygen lone pair interactions by the coadsorbed alkanes. In addition, we find evidence for the preferential adsorption of the alkane over the ether. For example, heptane is preferentially adsorbed over dibutyl ether even though it contains two fewer atoms in the molecular chain. This contrasts with the preferential adsorption of alcohols over alkanes reported previously (Messe, L.; Perdigon, A.; Clarke, S. M.; Inaba, A.; Arnold, T. Langmuir 2005, 21, 5085 5093). PMID- 17125352 TI - Structure and energy of Mo27SxCy clusters: a density functional theory study. AB - For understanding the carburization processes of MoSx catalysts, the structures and energies of Mo27SxCy cluster models have been computed at the level of density functional theory. The surface sulfur atoms on the Mo edge and S edge as well as bulky sulfur atoms have been replaced by atomic carbon, and the corresponding structures have S/C ratios in the range of 0.8-55. The formation of all Mo27SxCy structures is favored thermodynamically. It is also found that the formation of CS and C2 bridging units is more favored than the individual or separated replacements and that the formation of C2 bridging units is more favored than that of CS units. In contrast, the replacement of sulfur on the Mo edge is least favored. Furthermore, the replacement of the bulky sulfur on the Mo edge is equally favored as those of sulfur on the S edge. For aiding further experimental studies, the C=S and C=C stretching frequencies have been computed. PMID- 17125353 TI - Relevance in the Fischer-Tropsch synthesis of the formation of Fe-O-Ce interactions on iron-cerium mixed oxide systems. AB - A series of Fe-Ce mixed oxides (95 atom % Fe-5 atom % Ce) has been prepared by different methods: coprecipitation, impregnation, and physical mixture of Ce and Fe oxides. These solids have been tested in the Fischer-Tropsch synthesis. The characterization of the catalytic precursors was carried out by X-ray diffraction (XRD), Raman, Mossbauer, and X-ray photoelectron (XPS) spectroscopic techniques. When the preparation method ensures a microscopic contact between Fe and Ce cations in the solid, several types of Fe-Ce interactions are present in the calcined solids. The interactions take the shape of Fe-O-Ce bridges that can exist either in the hematite-like solid solution or in the interphase between the Fe oxide covered by microcrystals of Ce oxide. In the case of the hematite-like solid solution, Ce(IV) cations are dissolved in the alpha-Fe2O3 network. The promotion by Ce of the catalytic properties observed in the final catalysts can be directly related with the detection of these Fe-O-Ce bridges in the calcined solids. The Ce promotion results in a larger yield to hydrocarbons, a higher production of olefins, and a higher selectivity to medium and large chain hydrocarbons (larger than six carbon atoms). It is proposed that the Ce promotion is due to the presence of Fe0-Ce(III) ensembles in the final catalysts arising from the initial Fe-O-Ce bridges developed in the parent calcined samples. PMID- 17125354 TI - Photoluminescence spectral change in layered titanate oxide intercalated with hydrated Eu3+. AB - A number of interesting photoluminescence properties of titanate layered oxide intercalated with hydrated Eu3+ have been demonstrated. Photoluminescence intensity of Eu3+ decreased rapidly with time during irradiation by UV light having energy higher than the band gap energy of the host TiO (Ti(1.81)O4) layer. This is presumably due to the decrease in energy transfer from the host TiO layer to Eu3+ as a result of the change in the hydration state of water molecules surrounding Eu3+, which is caused by the hole produced in the TiO valence band. When irradiation was discontinued, the emission intensity gradually recovered. The recovery time increased when the water in the interlayer is removed by heat treatment. This indicates that the state of interlayer water changes during irradiation and returns to its initial state after discontinuation of irradiation. The excitation spectra changed drastically at any given wavelength upon irradiation with UV light. A comparison of the excitation spectra before and after irradiation reveals that only the excitation peak at around the irradiation wavelength decreased upon irradiation, as in the case of spectral hole burning. The hydration state of water molecules surrounding Eu3+ presumably changes depending on the irradiation wavelength, leading to the above spectral change because the Eu/TiO film has a superlattice structure producing holes with different energies. PMID- 17125355 TI - Raman imaging and Kelvin probe microscopy for the examination of the heterogeneity of doping in polycrystalline boron-doped diamond electrodes. AB - The issue of the heterogeneity of boron doping in microcrystalline diamond films was addressed by four different methods: micro-Raman spectroscopy and Raman imaging, Kelvin probe force microscopy, conducting atomic force microscopy, and scanning electrochemical microscopy. The samples were commercially available films from Windsor Scientific, with an average boron concentration of about 5 x 10(20) cm(-3). In agreement with previous works, all of the methods showed that the boron uptake was nonuniform across the surface of the electrode. Two different types of regions were evidenced, with metallic or semiconducting properties that were characterized with different types of Raman spectra. The line shape of these spectra was strongly dependent on the excitation wavelength. Local variations in electroactivity were evidenced by the SECM curves, which are related to the electronic properties of the individual grains, which, in turn, are governed by the boron content of the individual crystallites. In this study, two different micro-Raman imaging techniques were used that reveal the grain structure of the films: the images constructed from the diamond line intensity perfectly reproduced the optical image obtained by illuminating the sample in reflection. The method also allows detection of the presence of nondiamond carbon, especially in the metallic parts of the samples. Other spectral features (intensity of the boron-related broad lines, as well as the frequency and width of the diamond line) were used to construct images. In every case, the grain structure of the film was revealed, as well as twinning within individual crystallites. All approaches revealed that no enhanced doping or boron depletion occurred at the grain boundaries. PMID- 17125356 TI - Scanning tunneling microscopy of ethylated Si(111) surfaces prepared by a chlorination/alkylation process. AB - Scanning tunneling microscopy (STM) and computational modeling have been used to study the structure of ethyl-terminated Si(111) surfaces. The ethyl-terminated surface was prepared by treating the H-terminated Si(111) surface with PCl5 to form a Cl-terminated Si(111) surface with subsequent exposure to C(2)H(5)MgCl in tetrahydrofuran to produce an alkylated Si(111) surface. The STM data at 77 K revealed local, close-packed, and relatively ordered regions with a nearest neighbor spacing of 0.38 nm as well as disordered regions. The average spot density corresponded to approximately 85% of the density of Si atop sites on an unreconstructed Si(111) surface. Molecular dynamics simulations of a Si(111) surface randomly populated with ethyl groups to a total coverage of approximately 80% confirmed that the ethyl-terminated Si(111) surface, in theory, can assume reasonable packing arrangements to accommodate such a high surface coverage, which could be produced by an exoergic surface functionalization route such as the two-step chlorination/alkylation process. Hence, it is possible to consistently interpret the STM data within a model suggested by recent X-ray photoelectron spectroscopic data and infrared absorption data, which indicate that the two-step halogenation/alkylation method can provide a relatively high coverage of ethyl groups on Si(111) surfaces. PMID- 17125357 TI - First-principles study of K and Cs adsorbed on Pd(111). AB - The adsorptions of K and Cs on Pd(111) were studied by the density functional calculations within the generalized gradient approximation. The site preference, bonding character, work function, and electron structure of the system were analyzed. For K and Cs adsorption, the hcp hollow site was found to be preferred for all the coverages investigated. The calculated adsorption geometries for (2 x 2) and (square root 3 x square root 3)R30 degrees phases are both in reasonable agreement with the observed results. The decrease of the work function upon the adsorption of K and Cs can be attributed to a dipole moment associated with the polarized adsorbate atom, which is characterized by depletion of the electron charge in the alkali metal layer and a charge accumulation in the interface region. Our results indicate that the bonding of alkali metal with the Pd(111) surface has a mixed ionic and metallic bond character at low coverage and a metallic bond of covalent character at high coverage. PMID- 17125358 TI - Several types of bilayer smectic liquid crystals with ferroelectric and antiferroelectric properties in binary mixture of dimeric compounds. AB - The mesomorphic behavior and phase structure were examined in the mixture of two kinds of dimeric compounds, alpha,omega-bis(4-alkoxyanilinebenzylidene-4' carbonyloxy)pentane (mOAM5AMOm), by optical microscopy, X-ray diffraction, polarization switching, and second-harmonic generation measurements. One compound is 4OAM5AMO4 with a short terminal alkyl chain that forms a single-layer smectic phase (SmCAs) with a random mixing of spacer and tail groups. Another compound is 16OAM5AMO16 with a long terminal alkyl chain that forms a chiral, anticlinic, and antiferroelectric bilayer phase (SmCAb) with the bent molecules tilted to the bilayer. By mixing these two compounds, the SmCAs phase of 4OAM5AMO4 is easily destabilized, leading to the wide content region of the bilayer phases. In the bilayer regime, three other smectic phases are newly induced. Two of them are antiferroelectric and ferroelectric phases in which the molecules lie perpendicularly with respect to the layer. The other shows no polar response to an external electric field and behaves like a smectic A. The new appearance of these bilayer phases is discussed as a mixing effect of long and short tail groups. PMID- 17125359 TI - Growth mechanism of long aligned multiwall carbon nanotube arrays by water assisted chemical vapor deposition. AB - Highly aligned arrays of multiwalled carbon nanotube (MWCNT) on layered Si substrates have been synthesized by chemical vapor deposition (CVD). The effect of the substrate design and the process parameters on the growth mechanism were studied. Adding water vapor to the reaction gas mixture of hydrogen and ethylene enhanced the growth which led to synthesis of longer CNT arrays with high density. Environmental scanning electron microscopy (ESEM), energy-dispersive spectroscopy (EDS), and atomic force microscopy (AFM) were used to analyze the CNT morphology and composition. Quadrupole mass spectroscopy (QMS) provided in situ information on the gas spices within the reaction zone. On the basis of results, we verified the top growth mechanism and evaluated the reason of decline and stoppage of the CNT growth after extended period of deposition. Multilayered Si substrates with a top film of Al2O3, having appropriate roughness, provide favorable conditions to form catalyst islands with uniform distribution and size. Using water-assisted CVD process and optimized substrate design, our group succeeded to grow vertically aligned, patterned MWCNT up to 4-mm long. The arrays were of high purity and weak adhesion which allowed to be peeled off easily from the substrate. PMID- 17125360 TI - Significance of local density of states in the scanning tunneling microscopy imaging of alkanethiol self-assembled monolayers. AB - A systematic scanning tunneling microscopy (STM) study of alkanethiol self assembled monolayers (SAMs) is presented as a function of the bias voltage, tunneling current, and tip-termini separation. Stable and etch-pit free SAMs of close-packed undecanethiol/Au(111) were obtained after annealing in ultrahigh vacuum. STM revealed two distinct c(4x2) structures with four nonequivalent molecules per unit cell. For both structures, reversible contrast variations occur upon systematically tuning the bias voltage, the current, and the tip termini distance. These contrast transitions originate from probing the corresponding local density of states (LDOS) of each molecule and not from the reorientation of the alkanethiol chains. The STM contrast is particularly sensitive to the tip-termini separation in the range of 0.5-2.5 A, reflecting the distance-dependence of LDOS. At a fixed tip elevation, the STM contrast is less sensitive to changes in bias within 0.1-1.2 V. For the first time, we demonstrate that LDOS may override the physical height variations in the STM topographic contrast for alkanethiol SAM systems. PMID- 17125361 TI - Hydration and interfacial water in Nafion membrane probed by transmission infrared spectroscopy. AB - Infrared spectroscopy was applied to probe water inside pores and channels of Nafion membrane exchanged with either proton (H+) or sodium ions (Na+). Transmission measurements were performed on freestanding Nafion 112 (approximately 50 microm thickness) in a cell that enabled adjustment of the relative humidity. Experiments that employed Na+-exchanged Nafion focused on relative humidity environments at or below about 32% generated through the use of humectants. Under these conditions, narrow features in the O-H stretching spectral region near 3650-3720 cm(-1), previously attributed to interfacial water, were detected and matched to bands in vibrational sum frequency (VSF) spectra of water/air, water/organic, and salt-solution/air interfaces. The features correspond to the stretching mode of the "free" OH group of water oriented with one hydrogen atom toward other water molecules and interacting through hydrogen bonding and the other straddling the interface extending into fluorocarbon-rich regions (approximately 3668 cm(-1)) or air-filled segments (approximately 3700 cm(-1)) in the membrane. For membrane exchanged with H+, -SO3 groups were easily shifted to -SO3H as water was removed upon exposure to a few Torr of vacuum at 95 degrees C. In contrast, residual water was retained by membrane exchanged with Na+ after exposure to these conditions for up to 72 h. The permeation of methanol and acetone into Na+-exchanged Nafion 112 was also examined. The C-H and O-H stretching modes of methanol were perturbed in a manner that suggests the polymer disrupts hydrogen bonding interactions within the solvent, similar to the effect it exerts on pure water. For acetone, the C-H stretching modes were not shifted appreciably compared to those of the bulk liquid. However, the carbonyl band was affected, indicating the likely importance of dipolar interactions between solvent molecules and polar groups on the polymer. Control experiments performed with poly(hexafluoropropylene-co tetrafluoroethylene) (FEP) membrane did not show evidence for water or methanol permeation, which demonstrates the critical role played by the ion-filled channels and pores in facilitating solvent transport within Nafion membrane. PMID- 17125362 TI - Heterolytic splitting of H2 and CH4 on gamma-alumina as a structural probe for defect sites. AB - A combined use of DFT periodic calculations and spectroscopic studies (IR and solid-state NMR) shows that a gamma-alumina treated at 500 degrees C under high vacuum contains surface defects, which are very reactive toward H2 or CH4. The reaction of H2 on defect sites occurs at low temperature (ca. 25 degrees C) on two types of Al atoms of low coordination numbers, AlIII or AlIV, to give AlIV-H and AlV-H, respectively. The amount of defects as titrated by H2 at 25 and 150 degrees C is 0.043 and 0.069 site/nm2, respectively, in comparison with 4 OH/nm2). In contrast, CH4 reacts selectively at 100-150 degrees C on the most reactive AlIII sites to form the corresponding AlIV-CH3 (0.030 site/nm2). The difference of reactivity of H2 and CH4 is fully consistent with calculations (reaction and activation energy, DeltaE and DeltaE++). PMID- 17125363 TI - Computational study of location and role of fluoride in zeolite structures. AB - The distribution of fluoride ions has been studied in the pure silica IFR, ITH, IWR, STF and STT zeolite structures using computational techniques. The interactions between the F- and SDA+ ions (where SDA is the organic structure directing agent) are able to explain the F- cage occupation found experimentally. While studying the short-range fluoride-framework interactions, a relationship was found between the Si atoms forming the pentacoordinated units and the lowest F- defect energies, which rationalizes the experimental Si-F bonding in terms of energetic stability. It is proposed that the F- location is governed by a two step process. In a first stage, the electrostatic long-range forces and, especially, the interactions between the F- and the SDA+ ions, decide which cage will be filled with F-; in a second stage, once the F- cage location is decided, the F- forms a covalent bond with a Si site to form an energetically stable pentacoordinated unit [SiO4/2F]-. PMID- 17125364 TI - Mechanism of the oxidation-reduction of the MoVSbNbO catalyst: in operando X-ray absorption spectroscopy and electrical conductivity measurements. AB - The mechanism of the oxidation-reduction of the MoVSbNbO catalyst has been studied in dynamic conditions using X-ray absorption spectroscopy (XAS) and electrical conductivity measurements. XAS at Sb L1- and V/Mo K-edges permitted a better understanding of the chemical processes taking place in the M1 phase of the MoVSbNbO catalyst at different temperatures and atmosphere compositions. The reduction of antimony was already observed during the annealing of the M1 phase in He at 100 degrees C, which might be explained by the presence of hydrogen in the bronze-like structure of the M1 phase. Under operando conditions at 380 degrees C, we have found that Sb and V change their oxidation states depending on the C3H8/O2 ratio in the atmosphere. These changes occur simultaneously and with the same kinetics. Under the same conditions, variations in the oxidation state of Mo were not observed. These results prove that different types of oxygen (from the hexagonal channels and from the MO6 octahedral network) must be involved in the catalytic process although their relative contributions are different. It was found that the electrical conductance of the M1 phase correlates with the oxidation states of Sb and V and the concentration of oxygen vacancies. PMID- 17125365 TI - Influence of cation Na/Ca ratio on adsorption in LTA 5A: a systematic molecular simulation study of alkane chain length. AB - Recent adsorption isotherms of n-alkanes on Ca,Na-LTA-type zeolite afford development of a force field describing the interactions between calcium and n alkanes in configurational-bias Monte Carlo simulations. The force field of Calero et al. (J. Am. Chem. Soc. 2004, 126, 11377-11386) is able to accurately describe the adsorption properties of linear alkanes in the sodium form of FAU type zeolites. Here, we extend upon this type of force field by including calcium type ions. The force field was fitted to reproduce the calcium and sodium cations positions on LTA 5A and the experimental adsorption properties of n-alkanes over all range of temperatures and pressures. This opens up a vast amount of experimental data on LTA 5A, both on adsorption and diffusion. Furthermore, evaluation of half a century of reported n-alkane adsorption data on LTA-type zeolites indicates that there are many inconsistencies between the various data sets, possibly as a result of (i) undisclosed calcium and sodium contents, (ii) less than perfect drying of the hygroscopic zeolite, and (iii) coadsorption of contaminants such as vacuum grease. Having obtained our force field, and confirmed its reliability on predictions outside the calibration set, we apply the force field on two "open" problems: (a) the heats of adsorption and Henry coefficient as a function of chain length and (b) the effect of cations in LTA type zeolites. The molecular simulations shed new light on previous experimental findings, and we provide rationalizations on the molecular level that can be generalized to the class of cage/window-type nanoporous materials. PMID- 17125366 TI - Cathodoluminescence investigation of residual stress in Er3+:YAlO3 thin films grown on (110) SrTiO3 substrate by metal-organic chemical vapor deposition. AB - YAlO3 thin films doped with different amounts of Er3+ have been grown directly onto (110) SrTiO3 substrate using the metal-organic chemical vapor deposition method (MOCVD). X-ray diffraction patterns and the rocking curve of the (002) reflection point to the growth of <001>-oriented YAlO3 phase. Piezo-spectroscopic (PS) biaxial calibration was performed on two luminescence bands, related to transitions from the (4)S3/2 excited state, using a specially designed ball-on ring loading jig. Such a PS calibration allowed us to retrieve the rate of wavelength shift with stress without separating the grown film from the substrate. The outcome of the PS calibration has been applied to build up in the field emission scanning electron microscope (FEG-SEM) high-resolution maps of the residual stress field developed in the film. Results indicate that the residual stress in Er3+:YAlO3 films were compressive in nature and increased with increasing Er3+ dopant concentration. PMID- 17125367 TI - Gold films deposited over regular arrays of polystyrene nanospheres as highly effective SERS substrates from visible to NIR. AB - Gold nanostructured films of various thicknesses (15, 30, and 60 nm) are deposited over regular arrays of polystyrene nanospheres in an attempt to evaluate their potential as SERS-active substrates. Atomic force microscopy is used to topographically characterize the substrates as well as to ensure the thickness of the deposited gold films. The optical response of the prepared substrates recommends their use in SERS experiments with multiple laser lines from visible and NIR spectral domains. The assessment of the substrates' SERS activity is performed by using the 532, 633, and 830 nm excitation lines and different average enhancement factor (EF) values are obtained depending on the film thickness and employed laser line. The 60 nm gold nanostructured film generates the greatest local electromagnetic field confinement under NIR excitation and consequently gives rise to maximum SERS enhancement. The large tunability of surface plasmon excitation combined with the advantage of relatively high exhibited average EF values obtained under NIR excitation recommends these substrates as outstanding candidates for upcoming investigations of biological relevant molecules. PMID- 17125368 TI - Molecular dynamics simulation of liquid water confined inside graphite channels: dielectric and dynamical properties. AB - Electric and dielectric properties and microscopic dynamics of liquid water confined between graphite slabs are analyzed by means of molecular dynamics simulations for several graphite-graphite separations at ambient conditions. The electric potential across the interface shows oscillations due to water layering, and the overall potential drop is about -0.28 V. The total dielectric constant is larger than the corresponding value for the bulklike internal region of the system. This is mainly due to the preferential orientations of water nearest the graphite walls. Estimation of the capacitance of the system is reported, indicating large variations for the different adsorption layers. The main trend observed concerning water diffusion is 2-fold: on one hand, the overall diffusion of water is markedly smaller for the closest graphite-graphite separations, and on the other hand, water molecules diffuse in interfaces slightly slower than those in the bulklike internal areas. Molecular reorientational times are generally larger than those corresponding to those of unconstrained bulk water. The analysis of spectral densities revealed significant spectral shifts, compared to the bands in unconstrained water, in different frequency regions, and associated to confinement effects. These findings are important because of the scarce information available from experimental, theoretical, and computer simulation research into the dielectric and dynamical properties of confined water. PMID- 17125369 TI - Bistable electrical switching and memory effects in a thin film of copolymer containing electron donor-acceptor moieties and europium complexes. AB - A nonconjugated methacrylate copolymer (PCzOxEu) containing carbazole moieties (electron donors), 1,3,4-oxadiazole moieties (electron acceptors), and europium complexes in the pendant groups was synthesized via free radical copolymerization of methacrylate monomers containing the respective functional groups. The molecular structure and composition of PCzOxEu was characterized by elemental analysis, FT-IR, 1H NMR, 13C NMR, UV-vis absorption and fluorescence spectroscopies, gel permeation chromatography (GPC), X-ray photoelectron spectroscopy (XPS), and cyclic voltammetry (CyV). The resulting copolymer exhibited a relatively high glass transition temperature (Tg approximately 125 degrees C) and good solubility in common organic solvents. It could be cast into transparent films from solutions. For a thin film of PCzOxEu sandwiched between an indium-tin oxide (ITO) electrode and an Al electrode (ITO/PCzOxEu/Al), the structure behaved as a nonvolatile flash (rewritable) memory with accessible electronic states that could be written, read, and erased. The polymer memory exhibited an ON/OFF current ratio up to 10(5), switching response time of approximately 1.5 micros, more than 10(6) read cycles, retention time of more than 8 h, and write/erase voltages of about 4.4 V/-2.8 V under ambient conditions. The roles of oxadiazole moieties in improving the response time and retention time of the memory device were elucidated from the molecular simulation results. PMID- 17125370 TI - Thermal behavior of polyelectrolyte multilayer microcapsules: 2. Insight into molecular mechanisms for the PDADMAC/PSS system. AB - Polyelectrolyte multilayer capsules consisting of poly(diallyldimethylammonium chloride) (PDADMAC) and poly(styrene sulfonate) (PSS) were used as a model system to study the temperature-dependent behavior of polyelectrolyte multilayer films in aqueous media. Shells terminated with PSS shrink upon heating, whereas PDADMAC terminated ones swell, independent of the nature of the first layer, as measured by means of confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Elemental analysis shows that the initial exponential layer growth of the film leads to a nearly neutral overall charge in the first case or a high positive excess charge in the latter. Depending on this overall charge either surface tension, due to an unfavorable polymer-solvent interaction, or electrostatics dominates, resulting in a shrinkage or expansion of capsules, respectively. Thus, it is possible to swell temperature-shrunk capsules by coating them with an additional PDADMAC layer. Micro-DSC measurements prove that polyelectrolyte multilayers undergo a glass transition in water at which the wall material softens, allowing the rearrangements to occur. It is found that the thermal history has an influence on the temperature behavior of capsules, especially on those ones terminated with PDADMAC. Also, the molecular weight of the polyelectrolytes affects the rearrangement of capsules. The lower the molecular weight and thus the smaller the entanglement of chains, the easier polyelectrolytes can rearrange. PMID- 17125371 TI - Theoretical study of N-doped TiO2 rutile crystals. AB - The N-doping effects on the electronic and optical properties of TiO2 rutile crystal have been studied using density functional theory (DFT). The calculations of several possible N-doped structures show that band gaps have little reduction but some N 2p states lie within the band gap in the substitutional N to O structure and interstitial N-doped rutile supercell, which results in the reduction of the photon-transition energy and absorption of visible light. In contrast, substitutional N to Ti doped model has a significant band-gap narrowing. The results maybe clarify confusions in nitrogen-doped TiO2 rutile crystal. PMID- 17125372 TI - Chemical potential of a nonionic surfactant in solution. AB - The chemical potential of a surfactant in solution can be calculated from the Gibbs adsorption equation when the surface excess of the surfactant and the surface tension of the solution as a function of surfactant concentration are known. We have investigated a solution of the nonionic surfactant 1-palmitoyl-2 oleoyl-sn-glycero-3-phosphocholine (POPC) in the polar solvent 3 hydroxypropionitrile at concentrations below and above the critical micelle concentration (cmc). Neutral impact collision ion scattering spectroscopy was applied for the direct measurement of the surface excess of POPC as a function of concentration. The Gibbs adsorption equation was applied in conjunction with surface tension measurements to evaluate the chemical potential and the activity coefficients of POPC, respectively. We find that the solution shows ideal behavior up to the cmc and that the chemical potential remains constant at concentrations larger than the cmc. PMID- 17125373 TI - Preparation and characterization of copper-doped cobalt oxide electrodes. AB - Cobalt oxide (Co3O4) and copper-doped cobalt oxide (CuxCo(3-x)O4) films have been prepared onto titanium support by the thermal decomposition method. The electrodes have been characterized by different techniques such as cyclic voltammetry, scanning electron microscopy, X-ray diffraction, and X-ray photoelectron spectroscopy (XPS). The effect on the electrochemical and crystallographic properties and surface morphology of the amount of copper in the oxide layer has been analyzed. The XPS spectra correspond to a characteristic monophasic Cu-Co spinel oxides when x is below 1. However, when the copper content exceeds that for the stoichiometric CuCo2O4 spinel, a new CuO phase segregates at the surface. The analysis of the surface cation distribution indicates that Cu(II) has preference for octahedral sites. PMID- 17125374 TI - PdnCO (n = 1,2): accurate Ab initio bond energies, geometries, and dipole moments and the applicability of density functional theory for fuel cell modeling. AB - Electrode poisoning by CO is a major concern in fuel cells. As interest in applying computational methods to electrochemistry is increasing, it is important to understand the levels of theory required for reliable treatments of metal-CO interactions. In this paper we justify the use of relativistic effective core potentials for the treatment of PdCO and hence, by inference, for metal-CO interactions where the predominant bonding mechanism is charge transfer. We also sort out key issues involving basis sets and we recommend that bond energies of 17.2, 43.3, and 69.4 kcal/mol be used as the benchmark bond energy for dissociation of Pd2 into Pd atoms, PdCO into Pd and CO, and Pd2CO into Pd2 and CO, respectively. We calculated the dipole moments of PdCO and Pd2CO, and we recommend benchmark values of 2.49 and 2.81 D, respectively. Furthermore, we tested 27 density functionals for this system and found that only hybrid density functionals can qualitatively and quantitatively predict the nature of the sigma donation/pi-back-donation mechanism that is associated with the Pd-CO and Pd2-CO bonds. The most accurate density functionals for the systems tested in this paper are O3LYP, OLYP, PW6B95, and PBEh. PMID- 17125375 TI - Viologen-modified platinum clusters acting as an efficient catalyst in photocatalytic hydrogen evolution. AB - A highly efficient photocatalytic system for hydrogen evolution with dihydronicotinamide coenzyme (NADH) as a sacrificial agent in an aqueous solution has been constructed by using water-soluble platinum clusters functionalized with methyl viologen-alkanethiol (MVA2+) and a simple electron-donor dyad, 9-mesityl 10-methylacridinium ion (Acr+-Mes), which is capable of fast photoinduced electron transfer but extremely slow back electron transfer. The mean diameter of the platinum core was determined as R(CORE) = 1.9 nm with a standard deviation sigma = 0.5 nm by transmission electron microscopy (TEM). As a result, the hydrogen-evolution rate of the photocatalytic system with MVA2+-modified platinum clusters (MVA2+-PtC) is 10 times faster than the photocatalytic system with the mixture of the same amount of MVA2+ and platinum clusters as that of MVA2+-PtC under otherwise the same experimental conditions. The radical cation of NADH has been successfully detected by laser flash photolysis experiments. The decay of the absorbance due to NAD*, produced by the deprotonation from NADH*+, coincides with the appearance of the absorption band due to Acr*-Mes. This indicates electron transfer from NAD* to Acr+-Mes to give Acr*-Mes, which undergoes the electron-transfer reduction of MVA2+-PtC, leading to the efficient hydrogen evolution. PMID- 17125376 TI - Bismuth sulfide thin films with low resistivity on self-assembled monolayers. AB - Using self-assembled monolayers (SAMs), highly crystalline bismuth sulfide thin films with low electrical resistivity have been prepared from aqueous solution at low temperature (40-70 degrees C). The nucleation and growth process of Bi2S3 thin films was investigated in detail by XPS, AES, SEM, XRD, SAED, and HRTEM. Solution conditions have marked effects on the microstructure, growth rate, and mechanism of Bi2S3 films. Increased solution temperature resulted in a higher growth rate and a shorter induction time due to a higher supersaturation degree. In the solution of pH 1.12, homogeneous nucleation and the attachment process dominated the formation of Bi2S3 films. In contrast, at pH 0.47 Bi2S3 thin films were formed via heterogeneous nucleation and growth. The c-axial orientation of bismuthinite films was enhanced with the increase of reaction time. By controlling the solution supersaturation and reaction duration, highly crystalline Bi2S3 films composed of closely packed and coalescent crystallites could be realized, whose dark electrical resistivity could reach as low as 0.014 Omega cm without any post-treatment. PMID- 17125377 TI - Alkanethiol monolayers at reduced and oxidized zinc surfaces with corrosion protection: a sum frequency generation and electrochemistry investigation. AB - In this work, octadecanethiol (ODT) was demonstrated to form ordered monolayers at either electrochemically reduced or oxidized Zn surfaces, by means of sum frequency generation (SFG) spectroscopy, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). The SFG spectra of ODT-modified Zn electrodes featured three methyl group resonances in the C-H vibrational region (2800-3100 cm(-1)). A significant decrease in interfacial capacitance and an increase in charge-transfer resistance were observed in EIS measurement after ODT modification. The alkane chain tilt angle of ODT within a monolayer at the Zn surface was estimated as 0 degrees with respect to the surface normal by interfacial capacitance measurement via EIS. CV and SFG investigation revealed that ODT monolayers undergo reductive desorption from the Zn electrode in 0.5 M NaOH at -1.66 V (vs SCE) and in 0.5 M NaClO4 at -1.62 V. The integrated charge consumed to the desorption of ODT is determined as 87 mC/cm2 from the reductive peak on CV curve, resulting in a coverage of 9.0 x 10(-10) mol/cm2 (5.4 x 10(14) molecules/cm2) if assuming the reduction follows a one-electron process. ODT monolayers show corrosion protection to underlying zinc at the early immersion stage in base, salt, and acid media. However, the protection efficiency was reduced with immersion time due to the presence of defects within the monolayers. PMID- 17125378 TI - Mechanical simulation of the pressure and the relaxation to thermal equilibrium of a hot and dense rare gas cluster. AB - A cold atomic cluster can be very rapidly heated and compressed by a hypersonic impact at a hard surface. The impact can be simulated by computing a classical trajectory for the motion of the atoms. By suddenly confining the hot and dense cluster within a rigid container, it is possible to monitor the time evolution of the force acting on the faces of the container. It is found that the pressure computed this way very rapidly decays to a time-independent value. After a somewhat longer time, this value reproduces the value for the pressure computed as the sum of the kinetic and internal pressures. This agreement is expected for a system in equilibrium. These observations support the conclusion that there is a fast relaxation to thermal equilibrium in these essentially hard-sphere systems. The deviation from equilibrium is primarily due to the propagation of shock waves within the cluster. The equilibrium pressure can reach up to the megabar range. PMID- 17125379 TI - A fluctuation theory analysis of the salting-out effect. AB - An analysis of the salting-out, or Sechenow, effect is given in terms of Kirkwood Buff, or fluctuation, integrals. The analysis is formally exact but cannot easily be applied in its original form. When the solute that is being salted out is sparingly soluble, simplifications arise and the theory can be used to compute one of the Kirkwood-Buff integrals which is otherwise difficult to obtain. PMID- 17125380 TI - Predicting the phase behavior of nitrogen + n-alkanes for enhanced oil recovery from the SAFT-VR approach: examining the effect of the quadrupole moment. AB - The phase behavior of nitrogen + n-alkanes is studied within the framework of the statistical associating fluid theory for potentials of variable range (SAFT-VR). The effect of the quadrupole moment of nitrogen on the phase behavior is considered through an extension of the SAFT-VR equation that includes an additional contribution to the Helmholtz free energy due to quadrupolar interactions. A significant improvement in the description of the phase diagram of the binary mixtures of nitrogen with different n-alkanes is obtained with the new approach when compared to predictions from the original SAFT-VR EOS (i.e., without the quadrupolar term). The experimental value for the quadrupole moment of nitrogen is used in the new equation; thus, no additional parameters are employed. Given the nonideal nature of the binary mixtures, a binary interaction parameter is needed to describe the full-phase diagram and high-pressure critical lines of these systems; however, this can be fitted to a single system and successfully used to predict the phase behavior of other binary mixtures without additional fitting. Furthermore, only a single, transferable, cross-energy parameter is required when the quadrupolar term is considered, whereas a cross range parameter is also needed with the original SAFT-VR approach. The inclusion of the quadrupolar term in the equation of state therefore reduces the need to use effective parameters by explicitly including at the molecular level interactions due to the quadrupole moment. PMID- 17125381 TI - An automatic coarse-graining and fine-graining simulation method: application on polyethylene. AB - Multiscale modeling of a polymeric system is a challenging task in polymer physics. Here we introduce a bottom-up and then top-down scheme for the simulation of polyethylene (PE). The coarse-grained numerical potential for PE is derived through an automatic updating program by mapping its radial distribution function (RDF) from the Lowe-Andersen temperature controlling (LA) simulation onto the one from detailed molecular dynamics (MD) simulation. This coarse grained numerical potential can be applied in larger systems under the same thermodynamic conditions. We have tested the reliability of the derived potential in two ways. First, the blends of different linear low-density polyethylene (LLDPE) with high-density polyethylene (HDPE) have been simulated in LA with the coarse-grained numerical potentials and reasonable results are obtained. Moreover, Rouse scaling behavior is reproduced for monodispersed polymeric systems with different chain lengths. The atomistic details of the beads can be reintroduced into the coarse-grained HDPE and LLDPE/HDPE models, followed by a few MD runs to alleviate the local tension induced by this fine-graining procedure. The equilibrated large atomistic system can then be used for further studies. PMID- 17125382 TI - Molecular dynamics simulation of the cybotactic region in gas-expanded methanol carbon dioxide and acetone-carbon dioxide mixtures. AB - Local solvation and transport effects in gas-expanded liquids (GXLs) are reported based on molecular simulation. GXLs were found to exhibit local density enhancements similar to those seen in supercritical fluids, although less dramatic. This approach was used as an alternative to a multiphase atomistic model for these mixtures by utilizing experimental results to describe the necessary fixed conditions for a locally (quasi-) stable molecular dynamics model of the (single) GXL phase. The local anisotropic pair correlation function, orientational correlation functions, and diffusion rates are reported for two systems: CO2-expanded methanol and CO2-expanded acetone at 298 K and pressures up to 6 MPa. PMID- 17125383 TI - Discrete breathers for understanding reconstructive mineral processes at low temperatures. AB - Reconstructive transformations in layered silicates need a high temperature in order to be observed. However, very recently, some systems have been found where transformation can be studied at temperatures 600 degrees C below the lowest experimental results previously reported, including sol-gel methods. We explore the possible relation with the existence of intrinsic localized modes, known as discrete breathers. We construct a model for nonlinear vibrations within the cation layer, obtain their parameters, and calculate them numerically, obtaining their energies. Their statistics show that, although there are far less breathers than phonons, there are much more above the activation energy, making them good candidates to explain the reconstructive transformations at low temperatures. PMID- 17125384 TI - Molecular dynamics of apo-adenylate kinase: a distance replica exchange method for the free energy of conformational fluctuations. AB - A large domain motion in adenylate kinase from E. coli (AKE) is studied with molecular dynamics. AKE undergoes a large-scale rearrangement of its lid and AMP binding domains when the open form closes over its substrates, AMP, and Mg2+-ATP, whereby the AMP-binding and lid domains come closer to the core. The third domain, the core, is relatively stable during this motion. A reaction coordinate that monitors the distance between the AMP-binding and core domains is selected to be able to compare with the results of energy transfer experiments. Sampling along this reaction coordinate is carried out by using a distance replica exchange method (DREM), where systems that differ by a restraint potential enforcing different reaction coordinate values are independently simulated with periodic attempts at exchange of these systems. Several methods are used to study the efficiency and convergence properties of the DREM simulation and compared with an analogous non-DREM simulation. The DREM greatly accelerates the rate and extent of configurational sampling and leads to equilibrium sampling as measured by monitoring collective modes obtained from a principal coordinate analysis. The potential of mean force along the reaction coordinate reveals a rather flat region for distances from the open to a relatively closed AKE conformation. The potential of mean force for smaller distances has a distinct minimum that is quite close to that found in the closed form X-ray structure. In concert with a decrease in the reaction coordinate distance (AMP-binding-to-core distance) the lid-to-core distance of AKE also decreases. Therefore, apo AKE can fluctuate from its open form to conformations that are quite similar to its closed form X-ray structure, even in the absence of its substrates. PMID- 17125385 TI - Structural basis of fluorescence fluctuation dynamics of green fluorescent proteins in acidic environments. AB - Green fluorescent proteins (GFPs) have become powerful markers for numerous biological studies due to their robust fluorescence properties, site-specific labeling, pH sensitivity, and mutations for multiple-site labeling. Fluorescence correlation spectroscopy (FCS) studies have indicated that fluorescence blinking of anionic GFP mutants takes place on a time scale of 45-300 ms, depending on pH, and have been attributed to external proton transfer. Here we present experimental evidence indicating that conformational change in the protein &beta barrel is a determining step for the external protonation of GFP-S65T (at low pH) using time-resolved fluorescence and polarization anisotropy measurements. While the average anionic fluorescence lifetime of GFP-S65T is reduced by approximately 18% over a pH range of 3.6-10.0, the fluorescence polarization anisotropy decays mostly as a single exponential with a rotational time of phi = 17 +/- 1 ns, which indicates an intact beta-barrel with a hydrodynamic volume of 78 +/- 5 nm3. In contrast, the total fluorescence (525 +/- 50 nm) of the excited neutral state of S65T reveals a strong correlation between the fluorescence lifetime, structural conformation, and pH. The average fluorescence lifetime of the excited neutral state of S65T as a function of pH yields pKa approximately 5.9 in agreement with literature values using steady-state techniques. In contrast to the intact beta barrel at high pH, the anisotropy of neutral S65T (at pH 11), are found. C-8-deuteration of dGuo to give 8-D-dGuo removes the major proton hyperfine coupling at C-8. This isolates the anisotropic nitrogen couplings for each of the three species and aids our analyses. These anisotropic nitrogen couplings were assigned to specific nitrogen sites by use of 15N-substituted derivatives at N1, N2, and N3 atoms in dGuo. Both ESR and UV-visible spectra are reported for each of the species: G*+, G(-H)*, and G(-2H)*-. The experimental anisotropic ESR hyperfine couplings are compared to those obtained from DFT calculations for the various tautomers of G(-H)*. Using the B3LYP/6-31G(d) method, the geometries and energies of G*+ and its singly deprotonated state in its two tautomeric forms, G(N1-H)* and G(N2-H)*, were investigated. In a nonhydrated state, G(N2-H)* is found to be more stable than G(N1-H)*, but on hydration with seven water molecules G(N1-H)* is found to be more stable than G(N2-H)*. The theoretically calculated hyperfine coupling constants (HFCCs) of G*+, G(N1-H)*, and G(-2H)*- match the experimentally observed HFCCs best on hydration with seven or more waters. For G(-2H)*-, the hyperfine coupling constant (HFCC) at the exocyclic nitrogen atom (N2) is especially sensitive to the number of hydrating water molecules; good agreement with experiment is not obtained until nine or 10 waters of hydration are included. PMID- 17125390 TI - Photoexcitation of dinucleoside radical cations: a time-dependent density functional study. AB - The excited states of dinucleoside phosphates (dGpdG, dApdA, dApdT, TpdA, and dGpdT) in their cationic radical states were studied with time-dependent density functional theory (TD-DFT). The ground-state geometries of all the dinucleoside phosphate cation radicals considered, in their base stacked conformation, were optimized with the B3LYP/6-31G(d) method. Further, to take into account the effect of the aqueous environment surrounding the dinucleoside phosphates, the polarized continuum model (PCM) was considered and the excitation energies were computed by using the TD-B3LYP/6-31G(d) method. From this study, we find that the first transition in all the dinucleoside molecules involves hole transfer from base to base. dG*+pdG and dApdA*+ were found to have substantially lower first transition energies than others with two different DNA bases. Higher energy transitions involve base to sugar as well as base to base hole transfer. The calculated TD-B3LYP/6-31G(d) transition energies are in good agreement with previous calculations with CASSCF/CAS-PT2 level of theory. This TD-DFT work supports the experimental findings that sugar radicals formed upon photoexcitation of G*+ in gamma-irradiated DNA and suggests an explanation for the wavelength dependence found. PMID- 17125391 TI - Cation-pi Interactions and oxidative effects on Cu+ and Cu2+ binding to Phe, Tyr, Trp, and His amino acids in the gas phase. Insights from first-principles calculations. AB - The coordination properties of the four natural aromatic amino acids (AA(arom) = Phe, Tyr, Trp, and His) to Cu+ and Cu2+ have been exhaustively studied by means of ab initio calculations. For Cu+-Phe, Cu+-Tyr and Cu+-Trp, the two charge solvated tridentate N/O/ring and bidentate N/ring structures, with the metal cation interacting with the pi system of the ring, were found to be the lowest ones, relative DeltaG(298K) energies being less than 0.5 kcal/mol. The Cu+-His ground-state structure has the metal cation interacting with the NH2 group and the imidazole N. For these low-lying structures vibrational features are also discussed. Unlike Cu+ complexes, the ground-state structure of Cu2+-Phe, Cu2+ Tyr, and Cu2+-Trp does not present cation-pi interactions due to the oxidation of the aromatic ring induced by the metal cation. The ground-state structure of Cu2+ His does not present oxidation of the amino acid, the coordination to Cu2+ being tridentate with the oxygen of the carbonyl group, the nitrogen of the amine, and the N of the imidazole. Other less stable isomers, however, show oxidation of His, particularly of the imidazole ring, which can induce spontaneous proton transfer reactions from the NH of the imidazole to the NH2 of the backbone. Finally, the computed binding energies for Cu+-AA(arom) and Cu2+-AA(arom) systems have been computed, the order found for the single charged systems being Cu+-His > Cu+-Trp > Cu+-Tyr > Cu+-Phe, in very good agreement with the experimental data. PMID- 17125392 TI - Carotenoid radical cation formation in LH2 of purple bacteria: a quantum chemical study. AB - In LH2 complexes of Rhodobacter sphaeroides the formation of a carotenoid radical cation has recently been observed upon photoexcitation of the carotenoid S2 state. To shed more light onto the yet unknown molecular mechanism leading to carotenoid radical formation in LH2, the interactions between carotenoid and bacteriochlorophyll in LH2 are investigated by means of quantum chemical calculations for three different carotenoids--neurosporene, spheroidene, and spheroidenone--using time-dependent density functional theory. Crossings of the calculated potential energy curve of the electron transfer state with the bacteriochlorophyll Qx state and the carotenoid S1 and S2 states occur along an intermolecular distance coordinate for neurosporene and spheroidene, but for spheroidenone no crossing of the electron transfer state with the carotenoid S1 state could be found. By comparison with recent experiments where no formation of a spheroidenone radical cation has been observed, a molecular mechanism for carotenoid radical cation formation is proposed in which it is formed via a vibrationally excited carotenoid S1 or S*state. Arguments are given why the formation of the carotenoid radical cation does not proceed via the Qx, S2, or higher excited electron transfer states. PMID- 17125393 TI - How large is the [Fe(III)(protoporphyrin IX)]+ ion (hemin+) in the gas phase? AB - Comparison of the collision cross-section of the [Fe(III)-protoporphyrin IX]+ ion, hemin+, measured by means of ion-mobility experiments and the cross-sections calculated from theoretical structures based on density functional theory reveals that hemin+, in the gas phase, contains intramolecular hydrogen bonding between its two propionic acid side-chains. PMID- 17125394 TI - Penetration of beta-lactoglobulin into monoglyceride monolayers. dynamics, interactions, and topography of mixed films. AB - In this work we have analyzed the penetration of betalactoglobulin into a monoglyceride monolayer (monopalmitin or monoolein) spread at the air-water interface and its effects on the structural, dilatational, and topographical characteristics of mixed films. Dynamic tensiometry, surface film balance, Brewster angle microscopy (BAM), and surface dilatational rheology have been used, maintaining the temperature constant at 20 degrees C and the pH and ionic strength at 7 and 0.05 M, respectively. The initial surface pressure (mN/m) of the spread monoglyceride monolayer (pii(MONOGLYCERIDE)) at 10, 20, and the collapse point is the variable studied. Beta-lactoglobulin can penetrate into a spread monoglyceride monolayer at every surface pressure. The penetration of beta lactoglobulin into the monoglyceride monolayer with a more condensed structure, at the collapse point of the monoglyceride, requires monoglyceride molecular loss by collapse and/or desorption. However, the structural, topographical, and dilatational characteristics of monoglyceride penetrated by beta-lactoglobulin mixed monolayers are essentially dominated by the presence of monoglyceride (either monopalmitin or monoolein) in the mixed film. In fact, monoglyceride molecules have the capacity to re-enter the monolayer after expansion and recompression of the mixed monolayer. Thus, monoglyceride molecular loss by collapse and/or desorption is reversible. The topography of the monolayer under dynamic conditions corroborates these conclusions. PMID- 17125395 TI - Quantum chemical study on the coordination environment of the catalytic zinc ion in matrix metalloproteinases. AB - X-ray analyses of matrix metalloproteinases (MMPs) have shown that the catalytic zinc ion (Zn1) can bind to one to three water molecules in addition to three conserved histidine residues. To estimate the relative stability of the possible Zn1 coordination structures in the active site of the MMPs, we carry out computational analyses on the coordination environment of the Zn1 ion in the gelatinase A enzyme (or matrix metalloproteinase 2; MMP-2). Four-, five-, and six coordinated complexes representative of the Zn1 site are fully characterized by means of quantum mechanical (QM) methodologies. On one hand, B3LYP/LACVP* minimizations of various cluster models of the MMP-2 active site show that the trigonal bipyramidal geometry is energetically favored in the gas phase and that continuum solvent effects stabilize preferentially the tetrahedral complexes. On the other hand, B3LYP/OPLS-AA hybrid QM/molecular mechanical calculations in the solvated catalytic domain of the MMP-2 enzyme complemented with electrostatic Poisson-Boltzmann calculations show that the mature enzyme presents most likely a Zn1 ion coordinated by three histidine residues and two water molecules, while the active site glutamic acid is negatively charged. In consonance with X-ray diffraction data, other possible Zn1 configurations, a six-coordinated structure with Zn1-water as well as four- and five-coordinated complexes with a Zn1-bound hydroxide, are predicted to be very close in energy. PMID- 17125396 TI - Probing the behavior of confined water by proton-transfer reactions. AB - The picosecond dynamics of a bifunctional and H-bonding molecule, 7 hydroxyquinoline (7HQ), has been studied in a reverse micelle with increasing water content. The fluorescence kinetics has a complex behavior as the water content is changed. All reactions are irreversible, and a two-step mechanism is invoked to explain the observations. H2O/D2O exchange and excitation energy effects show that the second step has a higher barrier and that the corresponding reaction occurs through tunneling. The results clearly indicate two regimes of water nanopool behavior switching at W0 approximately 5 (W0 = [water]/[surfactant]). Water collective dynamics explains these observations. The lower fluidity of confined water within the reverse micelle with respect to normal bulk water alters the related H-bond network dynamics and therefore is responsible for the slower proton-transfer processes. PMID- 17125397 TI - Collective fluctuations in ordered fluids investigated by two-dimensional electron-electron double resonance spectroscopy. AB - Two-dimensional electron-electron double resonance (2D-ELDOR) is a technique that is sensitive to the dynamical processes affecting spin labels in complex fluid environments. In ordered fluids, such as membrane vesicles, the 2D-ELDOR experiment is affected by the molecular tumbling in the locally ordered environment. This motion occurs on two different time scales, the faster molecular motion relative to the local director, and the slower collective fluctuations of the director field. In the experimental study of Patyal, Crepeau, and Freed (Biophys. J. 1997, 73, 2201), it was found that the widths of the autopeaks of the 2D-ELDOR spectrum increased as a function of the mixing time. In the present work, a theory is developed for the effects of director fluctuations on the autopeaks in the 2D-ELDOR experiment by employing an analytical solution of the stochastic Liouville equation for which the director field is treated as a multidimensional Gaussian process, as previously developed by Frezzato, Kothe, and Moro (J. Phys. Chem. B 2001, 105, 1281 and J. Phys. Chem. B 2004, 108, 9505). Good agreement is found between theory and experiment, notably the only adjustable parameter is k, the bending elastic modulus of the membrane. The values of k = 11 x 10(-20) J for 1,2-dipalmitoyl-sn-glycero-phosphatidylcholine (DPPC) vesicles and k = 15 x 10(-20) J for DPPC/gramicidin A (5:1) vesicles, both at 45 degrees C, were found from the analysis and agree well with previous related measurements by other physical techniques. This establishes 2D-ELDOR as a useful technique to study the elastic properties of biological membranes. PMID- 17125398 TI - Collective dynamics of lysozyme in water: terahertz absorption spectroscopy and comparison with theory. AB - To directly measure the low-frequency vibrational modes of proteins in biologically relevant water environment rather than previously explored dry or slightly hydrated phase, we have developed a broadband terahertz spectrometer suitable for strongly attenuating protein solutions. Radiation is provided by harmonic multipliers (up to 0.21 THz), a Gunn oscillator (at 0.139 THz), and the UCSB free-electron lasers (up to 4.8 THz). Our spectrometer combines these intense sources with a sensitive cryogenic detector and a variable path length sample cell to detect radiation after it is attenuated by more than 7 orders of magnitudes by the aqueous sample. Using this spectrometer, we have measured the molar extinction of solvated lysozyme between 0.075 and 3.72 THz (2.5-124 cm( 1)), and we made direct comparison to several published theoretical models based on molecular dynamics simulations and normal-mode analysis. We confirm the existence of dense, overlapping normal modes in the terahertz frequency range. Our observed spectrum, while in rough qualitative agreement with these models, differs in detail. Further, we observe a low-frequency cutoff in terahertz dynamics between 0.2 and 0.3 THz, and we see no evidence of a predicted normal mode at approximately 0.09 THz for the protein. PMID- 17125399 TI - Correlation of polarizabilities with Van der Waals interactions in pi-systems. AB - This work aims to (i) provide a semiquantitative relationship that can be used to estimate the binding energy, equilibrium separation, and potential energy surface (PES) for supermolecules consisting of benzene and small polycyclic aromatic hydrocarbons (PAHs) in parallel configuration and (ii) give a qualitative description of pi-pi interactions between PAHs. We compute the one-dimensional PES of benzene translated parallel to various PAHs within the framework of second order Moller-Plesset (MP2) perturbation theory. For PAHs of small MW difference, we observe a linear correlation between the binding energy and the number of carbon atoms in the supermolecule. The PES of these supermolecules is fit to an (exp-6) function whose variables are subsequently used to derive a mass-centered potential energy function as a function of the number of carbon atoms in the supermolecule. The linear dependence of the binding energy in the supermolecular series examined here can be directly correlated to the average polarizability product of the supermolecule. Last, we consider the supermolecular series of benzene with n-polyacenes to study the convergence of pi-pi interactions between PAHs when their size is considerably different. PMID- 17125400 TI - Li+ transport in lithium sulfonylimide-oligo(ethylene oxide) ionic liquids and oligo(ethylene oxide) doped with LiTFSI. AB - The Li+ environment and transport in an ionic liquid (IL) comprised of Li+ and an anion of bis(trifluoromethanesulfonyl)imide anion (TFSI-) tethered to oligoethylene oxide (EO) (EO(12)TFSI-/Li+) were determined and compared to those in a binary solution of the oligoethylene oxide with LiTFSI salt (EO(12)/LiTFSI) by using molecular dynamics (MD) simulations and AC conductivity measurements. The latter revealed that the AC conductivity is 1 to 2 orders of magnitude less in the IL compared to the oligoether/salt binary electrolyte with greater differences being observed at lower temperatures. The conductivity of these electrolytes was accurately predicted by MD simulations, which were used in conjunction with a microscopic model to determine mechanisms of Li+ transport. It was discerned that structure-diffusion of the Li+ cation in the binary electrolyte (EO(12)/LiTFSI-) was similar to that in EO(12)TFSI-/Li+ IL at high temperature (>363 K), thus, one can estimate conductivity of IL at this temperature range if one knows the structure-diffusion of Li+ in the binary electrolyte. However, the rate of structure-diffusion of Li+ in IL was found to slow more dramatically with decreasing temperature than in the binary electrolyte. Lithium motion together with EO(12) solvent accounted for 90% of Li+ transport in EO(12)/LiTFSI-, while the Li+ motion together with the EO(12)TFSI- anion contributed approximately half to the total Li+ transport but did not contribute to the charge transport in IL. PMID- 17125403 TI - High-throughput in vitro profiling assays: lessons learnt from experiences at Novartis. AB - This article reviews the use of a selection of in vitro assays implemented at Novartis and intends to address exposure and safety in early drug discovery. The authors' own experience, based on a large number of 'real' drug discovery compounds, is described to reflect on what has worked, where improvement is needed and how to best use the data for decision making. Possible strategies are discussed, and guidelines are provided on how to organise assays, extract value and contribute knowledge from the data. PMID- 17125404 TI - Investigations toward enhanced understanding of hepatic idiosyncratic drug reactions. AB - Idiosyncratic drug reactions (IDRs) of a hepatic origin are a major health concern and a notoriously difficult challenge for the pharmaceutical industry. These types of adverse events are rare, with a typical occurrence of 1 in 100 to 1 in 100,000 patients. Typical adverse outcomes are most likely statistically impossible to predict in traditional preclinical safety studies or clinical trials. Unfortunately, these reactions can pose a significant risk to the public health, resulting in devastating consequences such as irreversible liver injury, liver transplantation and fatality. This review provides many examples of experimental efforts that are underway for a better understanding of molecular events that may be responsible for IDRs. A list of existing hypotheses for IDRs is also provided, each with current literature examples or supporting evidence. The possibilities for developing suitable animal models for the prediction and characterisation of IDRs are elaborated, especially for a drug-inflammation interaction rat model of hepatic IDR. The need for predictive biomarkers of IDR is addressed, with the exploration of some possible candidates. Finally, the use of primary human hepatocyte culture systems is explored as an in vitro system, with application for providing an increased mechanistic knowledge of IDR. Several examples of informative studies on the nature of IDRs that employ toxicogenomic and proteomic technologies are summarised. PMID- 17125405 TI - Pregnane X receptor and natural products: beyond drug-drug interactions. AB - The pregnane X receptor (PXR, NR1I2) is a member of the nuclear receptor superfamily that is activated by a myriad of compounds and natural products in clinical use. Activation of PXR represents the basis for several clinically important drug-drug interactions. Although PXR activation has undesirable effects in patients on combination therapy, it also mediates the hepatoprotective effects exhibited by some herbal remedies. This review focuses on PXR activation by natural products and the potential therapeutic opportunities presented. In particular, the biological effects of St. John's Wort, gugulipid, kava kava, Coleus forskolii, Hypoxis, Sutherlandia, qing hao, wu wei zi, gan cao and other natural products are discussed. The impact of these natural products on drug metabolism and hepatoprotection is highlighted in the context of activation and antagonism of PXR. PMID- 17125406 TI - Cross-species comparative toxicogenomics as an aid to safety assessment. AB - Cross-species comparative toxicogenomics has the potential for improving the understanding of the different responses of animal models to toxicants at a molecular level. This understanding could then lead to a more accurate extrapolation of the risk posed by these toxicants to humans. Cross-species comparative studies have been carried out at the genomic sequence level and using microarrays to examine changes in global mRNA profiles. However, these studies face considerable bioinformatic challenges in terms of identifying which genes are truly orthologous across species. The resources to analyse such studies, in the context of such orthologues, beg improvement. Finally, the experimental design of such studies needs to be carefully considered to make their results fully interpretable. These issues are discussed, along with the current state-of the-art cross-species comparative toxicogenomics in this review. PMID- 17125407 TI - Species differences between mouse, rat, dog, monkey and human CYP-mediated drug metabolism, inhibition and induction. AB - Animal models are commonly used in the preclinical development of new drugs to predict the metabolic behaviour of new compounds in humans. It is, however, important to realise that humans differ from animals with regards to isoform composition, expression and catalytic activities of drug-metabolising enzymes. In this review the authors describe similarities and differences in this respect among the different species, including man. This may be helpful for drug researchers to choose the most relevant animal species in which the metabolism of a compound can be studied for extrapolating the results to humans. The authors focus on CYPs, which are the main enzymes involved in numerous oxidative reactions and often play a critical role in the metabolism and pharmacokinetics of xenobiotics. In addition, induction and inhibition of CYPs are compared among species. The authors conclude that CYP2E1 shows no large differences between species, and extrapolation between species appears to hold quite well. In contrast, the species-specific isoforms of CYP1A, -2C, -2D and -3A show appreciable interspecies differences in terms of catalytic activity and some caution should be applied when extrapolating metabolism data from animal models to humans. PMID- 17125408 TI - Involvement of enzymes other than CYPs in the oxidative metabolism of xenobiotics. AB - Although the majority of oxidative metabolic reactions are mediated by the CYP superfamily of enzymes, non-CYP-mediated oxidative reactions can play an important role in the metabolism of xenobiotics. The (major) oxidative enzymes, other than CYPs, involved in the metabolism of drugs and other xenobiotics are: the flavin-containing monooxygenases, the molybdenum hydroxylases (aldehyde oxidase and xanthine oxidase), the prostaglandin H synthase, the lipoxygenases, the amine oxidases (monoamine, polyamine, diamine and semicarbazide-sensitive amine oxidases) and the alcohol and aldehyde dehydrogenases. In a similar manner to CYPs, these oxidative enzymes can also produce therapeutically active metabolites and reactive/toxic metabolites, modulate the efficacy of therapeutically active drugs or contribute to detoxification. Many of them have been shown to be important in endobiotic metabolism, and, consequently, interactions between drugs and endogenous compounds might occur when they are involved in drug metabolism. In general, most non-CYP oxidative enzymes appear to be noninducible or much less inducible than the CYP system, although some of them may be as inducible as some CYPs. Some of these oxidative enzymes exhibit polymorphic expression, as do some CYPs. It is possible that the contribution of non-CYP oxidative enzymes to the overall metabolism of xenobiotics is underestimated, as most investigations of drug metabolism in discovery and lead optimisation are performed using in vitro test systems optimised for CYP activity. PMID- 17125409 TI - Chemical toxicology: reactive intermediates and their role in pharmacology and toxicology. AB - Reactive intermediates formed during the metabolism of drugs have been investigated extensively over the past decades. Today, interest in reactive intermediates in drug discovery is focused on minimising bioactivation in hopes of reducing the risk of causing so-called idiosyncratic toxicity. These efforts are justified based on the 'hapten hypothesis', namely, that on binding to protein, reactive intermediates may elicit an immune response to the modified protein, leading to a cascade of events that ultimately manifests as a toxic outcome. However, the pharmacological action of certain drugs depends on reactive intermediates that modify critical amino acid residues of proteins, typically enzymes, thereby altering their activity. Thus, the notion that reactive intermediates are inherently dangerous is unjustified. When a reactive intermediate is necessary for the desired pharmacological effect of a drug, the selectivity it displays towards the target protein is crucial, as off-target binding may produce unwanted toxicities. On the other hand, reactive intermediates may play no role in toxicity. This review provides a balanced perspective, primarily focusing on the proposed role of reactive intermediates in drug toxicity, while also highlighting examples in which they are involved in causing the desired pharmacology. It is hoped that this knowledge can help scientists involved in drug discovery and development in their challenging task of producing safe and effective drugs. PMID- 17125410 TI - Using pharmacokinetics to predict the effects of pregnancy and maternal-infant transfer of drugs during lactation. AB - Knowledge of pharmacokinetics and the use of a mechanistic-based approach can improve our ability to predict the effects of pregnancy for medications when data are limited. Despite the many physiological changes that occur during pregnancy that could theoretically affect absorption, bioavailability does not appear to be altered. Decreased albumin and alpha(1)-acid glycoprotein concentrations during pregnancy will result in decreased protein binding for highly bound drugs. For drugs metabolised by the liver, this can result in misinterpretation of total plasma concentrations of low extraction ratio drugs and overdosing of high extraction ratio drugs administered by non-oral routes. Renal clearance and the activity of the CYP isozymes, CYP3A4, 2D6 and 2C9, and uridine 5'-diphosphate glucuronosyltransferase are increased during pregnancy. In contrast, CYP1A2 and 2C19 activity is decreased. The dose of a drug an infant receives during breastfeeding is dependent on the amount excreted into the breast milk, the daily volume of milk ingested and the average plasma concentration of the mother. The lipophilicity, protein binding and ionisation properties of a drug will determine how much is excreted into the breast milk. The milk to plasma concentration ratio has large inter- and intrasubject variability and is often not known. In contrast, protein binding is usually known. An extensive literature review was done to identify case reports including infant concentrations from breast-fed infants exposed to maternal drugs. For drugs that were at least 85% protein bound, measurable concentrations of drug in the infant did not occur if there was no placental exposure immediately prior to or during delivery. Knowledge of the protein binding properties of a drug can provide a quick and easy tool to estimate exposure of an infant to medication from breastfeeding. PMID- 17125411 TI - Pharmacokinetics and interactions of headache medications, part I: introduction, pharmacokinetics, metabolism and acute treatments. AB - Recent progress in the treatment of primary headaches has made available specific, effective and safe medications for these disorders, which are widely spread among the general population. One of the negative consequences of this undoubtedly positive progress is the risk of drug-drug interactions. This review is the first in a two-part series on pharmacokinetic drug-drug interactions of headache medications. Part I addresses acute treatments. Part II focuses on prophylactic treatments. The overall aim of this series is to increase the awareness of physicians, either primary care providers or specialists, regarding this topic. Pharmacokinetic drug-drug interactions of major severity involving acute medications are a minority among those reported in literature. The main drug combinations to avoid are: i) NSAIDs plus drugs with a narrow therapeutic range (i.e., digoxin, methotrexate, etc.); ii) sumatriptan, rizatriptan or zolmitriptan plus monoamine oxidase inhibitors; iii) substrates and inhibitors of CYP2D6 (i.e., chlorpromazine, metoclopramide, etc.) and -3A4 (i.e., ergot derivatives, eletriptan, etc.), as well as other substrates or inhibitors of the same CYP isoenzymes. The risk of having clinically significant pharmacokinetic drug-drug interactions seems to be limited in patients with low frequency headaches, but could be higher in chronic headache sufferers with medication overuse. PMID- 17125412 TI - Pharmacokinetics and interactions of headache medications, part II: prophylactic treatments. AB - The present part II review highlights pharmacokinetic drug-drug interactions (excluding those of minor severity) of medications used in prophylactic treatment of the main primary headaches (migraine, tension-type and cluster headache). The principles of pharmacokinetics and metabolism, and the interactions of medications for acute treatment are examined in part I. The overall goal of this series of two reviews is to increase the awareness of physicians, primary care providers and specialists regarding pharmacokinetic drug-drug interactions (DDIs) of headache medications. The aim of prophylactic treatment is to reduce the frequency of headache attacks using beta-blockers, calcium-channel blockers, antidepressants, antiepileptics, lithium, serotonin antagonists, corticosteroids and muscle relaxants, which must be taken daily for long periods. During treatment the patient often continues to take symptomatic drugs for the attack, and may need other medications for associated or new-onset illnesses. DDIs can, therefore, occur. As a whole, DDIs of clinical relevance concerning prophylactic drugs are a limited number. Their effects can be prevented by starting the treatment with low dosages, which should be gradually increased depending on response and side effects, while frequently monitoring the patient and plasma levels of other possible coadministered drugs with a narrow therapeutic range. Most headache medications are substrates of CYP2D6 (e.g., beta-blockers, antidepressants) or CYP3A4 (e.g., calcium-channel blockers, selective serotonin re-uptake inhibitors, corticosteroids). The inducers and, especially, the inhibitors of these isoenzymes should be carefully coadministered. PMID- 17125413 TI - Pharmacokinetics, toxicology and safety of lamotrigine in epilepsy. AB - Optimisation of pharmacotherapy for epilepsy requires consideration of the impact of drug metabolism and toxicology on the therapeutic profiles and clinical use of antiepileptic drugs (AEDs). This review discusses the pharmacokinetics and toxicology of the AED lamotrigine, and considers the implications of these data for optimising its use in the management of epilepsy. Lamotrigine has good absorption, minimal plasma protein binding and linear pharmacokinetics. Partly because of these properties, frequent dosing adjustments are generally unnecessary, and therapeutic monitoring is not required under most circumstances. Lamotrigine is not associated with clinically significant neurological, cognitive, metabolic, hepatic or reproductive endocrine toxicity. Like other AEDs, including carbamazepine and phenytoin, lamotrigine has been associated with serious rash. With some exceptions, lamotrigine has relatively few clinically relevant drug interactions, a characteristic important in reducing safety risks, especially among patients who require polytherapy. The clinical impact of pharmacokinetic interactions between lamotrigine and enzyme-inducing AEDs or valproate can be minimised by adhering to recommended dose-escalation schedules with demonstrated reliability in clinical trials and clinical practice. Likewise, adhering to recommended dosing guidelines can minimise the risk of lamotrigine associated rash. The pharmacokinetic, toxicology and safety profiles of lamotrigine make the drug suitable for use across a spectrum of patients with epilepsy. PMID- 17125414 TI - Glatiramer acetate therapy for multiple sclerosis: a review. AB - The past decade has witnessed a revolution in the treatment of multiple sclerosis (MS), the most common demyelinating disorder of the human CNS. After being considered as an untreatable disease for more than a century, six disease modifying treatments have been approved between 1993 and 2006. Glatiramer acetate (GA) is a worldwide drug approved for the treatment of relapsing-remitting MS in 1996. The drug is a synthetic copolymer of four amino acids based on the composition of myelin basic protein, one of several putative autoantigens implicated in the pathogenesis of MS. Three separate double-blind, placebo controlled trials have established its efficacy in relapsing-remitting MS. Observations from an ongoing study, the longest prospective study in MS therapeutics so far, suggest that the effect of GA in reducing the relapse rate and neurological disability is maintained over a 10-year period. Independent investigators have identified several putative immunological mechanisms of action of GA, with the unique observation of the generation of GA-reactive T-helper 2 (anti-inflammatory) polarised lymphocytes within days to weeks of initiating therapy and sustaining an anti-inflammatory milieu for years in the peripheral immune system and, presumably, in the CNS. Emerging data from immunological and imaging studies quantifying axonal injury in the brain point towards neuroprotective abilities of GA. Combined with its remarkable safety and tolerability, long-term efficacy and neuroprotective effect, GA presents it self as a first-line choice in relapsing-remitting MS, and holds immense promise in developing its potential as a combination therapy in MS, as well as extending its indications to other neurodegenerative diseases. PMID- 17125416 TI - Toward optimal health: Robert Kushner, M.D., offers a practical approach to assessment of overweight patients. PMID- 17125417 TI - The changing face of women with disabilities: are we ready? PMID- 17125418 TI - Racial/Ethnic and socioeconomic differences in multiple risk factors for heart disease and stroke in women: behavioral risk factor surveillance system, 2003. AB - OBJECTIVE: To examine racial/ethnic and socioeconomic disparities in multiple risk factors for heart disease and stroke among women. METHODS: Data from 153,466 adult women in the 2003 Behavioral Risk Factor Surveillance System (BRFSS), a telephone survey of U.S. adults, were used to assess the prevalence of multiple (i.e., >or=2 of diabetes, current smoking, high blood pressure, high cholesterol, obesity, or physical inactivity) risk factors for heart disease and stroke. Descriptive and multivariable analyses assessed differences in multiple risk factors among racial/ethnic and socioeconomic groups. RESULTS: More than one third (36.5%) of all women had multiple risk factors. The age-standardized prevalence of multiple risk factors was lowest in whites and Asians. After adjustment for age, income, education, and health coverage, the odds for multiple risk factors was greater in black (OR = 1.53, 95% CI = 1.42-1.64) and Native American women (1.36, 95% CI = 1.11-1.67) and lower for Hispanic women (OR = 0.83, 95% CI = 0.76-0.91) compared with white women. Prevalence estimates and odds of multiple risk factors increased with age; decreased with education, income, and employment; and were lower in those with no health coverage. Smoking was more common in younger women, whereas older women were more likely to have medical conditions (high blood pressure, high cholesterol, or diabetes) and be physically inactive. CONCLUSIONS: Over one third of U.S. women have two or more risk factors for heart disease and stroke. Prevention programs that target risk reduction are especially critical to decrease the burden of heart disease and stroke in these higher-risk U.S. women. PMID- 17125419 TI - Hyperlipidemia guideline adherence and association with patient gender. AB - BACKGROUND: Gender disparities in cardiovascular disease (CVD) management have become increasingly apparent in recent years. Previous research has focused on inpatient disparities, but little is known about how patient gender affects assessment, treatment, and management of patients for hyperlipidemia and cardiovascular risk in primary care settings. Patients with coronary artery disease (CAD) and hyperlipidemia are at high risk for cardiovascular and cerebrovascular morbidity. We sought to examine the effect of patient gender on assessment, treatment, and target maintenance of hyperlipidemia among patients with CAD in a primary care setting. METHODS: Chart abstraction was done for 715 patients with CAD in 55 family practices in New Jersey and eastern Pennsylvania as part of the Using Learning Teams for Reflective Adaptation (ULTRA) project. Hyperlipidemia assessment, treatment, and target adherence scores were determined for those at-risk patients based on National Heart, Lung, and Blood Institute (NHLBI) recommended National Cholesterol Education Program (NCEP) ATP III guidelines. Generalized linear models were used to determine the association of hyperlipidemia guideline adherence with patient gender, using comorbidities and age as confounders. RESULTS: After controlling for comorbidities and age, women were less likely to be assessed for lipids (p = 0.0462). There was no difference in treatment (p = 0.1074) or target laboratory values (p = 0.3949). CONCLUSIONS: Women with CAD are less often assessed for lipids than men in primary care practices. More intensive efforts may be necessary to educate physicians and patients about cardiovascular risk for women. PMID- 17125420 TI - Quality of life and menopause in women with physical disabilities. AB - OBJECTIVE: The goal of this cross-sectional study was to explore quality of life (QOL) in a sample of postmenopausal women with physical disabilities due to polio contracted in childhood. A structural equation model was used to confirm that menopause symptoms will have a minimal effect on QOL when disability-related variables are taken into account. METHODS: A sample of 752 women who were postmenopausal completed a written survey. The structural equation model contained two measured predictors (age, severity of postpolio sequelae) and one latent predictor (menopause symptoms defined by four measured indicators). Functional status (defined by two measured indicators) was included as a mediator, with QOL (defined by three measured indicators) as the outcome. RESULTS: The original model yielded acceptable fit indices (CFI = 0.96, RMSEA = 0.055) but resulted in a number of unexpected relationships that proved to be artifacts after model respecification. The respecified model yielded a nonsignificant chi-square value, which indicated no significant discrepancy between the proposed model and the observed data (chisquare = 18.5, df = 13, p = 0.138). All fit indices indicated a good fit: CFI = 0.997, NNFI = 0.987, chi square/df = 1.43, and RMSEA = 0.024. CONCLUSIONS: When the effects of postpolio sequelae and functional status are included in the structural equation model, only the psychological symptoms of menopause play a prominent role in explaining QOL in this sample. The clinical implications of these findings suggest that attention to psychological symptoms and an exclusive focus on the physical aspects of menopause to the exclusion of other midlife life stressors and influences on a woman's psychological well-being ignore the larger context of life in which they live. In particular, many women with disabilities may contend with additional or exacerbated stressors related to their disability. PMID- 17125421 TI - The association between body mass index and osteoporosis in patients referred for a bone mineral density examination. AB - PURPOSE: Osteoporosis affects 4-6 million (13%-18%) postmenopausal white women in the United States. Most studies to date on risk factors for osteoporosis have considered body mass index (BMI) only as a possible confounder. In this study, we assess the direct relationship between BMI and osteoporosis. METHODS: We conducted a cross-sectional study among women aged 50-84 years referred by their physicians for a bone mineral density (BMD) examination at Baystate Medical Center between October 1998 and September 2000. BMI was determined prior to the BMD examination in the clinic. Information on other risk factors was obtained through a mailed questionnaire. Ordinal logistic regression was used to model the association between BMI and osteoporosis, controlling for confounding factors. RESULTS: BMI was inversely associated with BMD status. After adjustment for age, prior hormone replacement therapy (HRT) use, and other factors, odds ratios (OR) for low, high, and obese compared with moderate BMI women were 1.8 (95% CI 1.2 2.7), 0.46 (95% CI 0.29- 0.71), and 0.22 (95% CI 0.14-0.36), respectively, with a significant linear trend (p < 0.0001) across BMI categories. Evaluating BMI as a continuous variable, the odds of bone loss decreased 12% for each unit increase in BMI (OR = 0.88, 95% CI 0.85-0.91). CONCLUSIONS: Women with low BMI are at increased risk of osteoporosis. The change in risk associated with a 1 unit change in BMI ( approximately 5-8 lb) is of greater magnitude than most other modifiable risk factors. To help reduce the risk of osteoporosis, patients should be advised to maintain a normal weight. PMID- 17125422 TI - Women's health issues with fibromyalgia syndrome. AB - BACKGROUND: Fibromyalgia syndrome (FMS) involves multiple sensory, somatic, and cognitive symptoms that are bound to affect or be affected by physical and mental health status and behavioral components of daily life. METHODS: From a telephone survey of 442 women with and 205 women without FMS as volunteers, data were compared on (1) general health status, (2) reproductive and sleep-related diagnoses, and (3) lifestyle health behaviors. RESULTS: All multiple or logistics regression analyses for group differences were controlled for age, body mass index (BMI), race, employment status, marital status, having a college degree, low household income, and having ever been diagnosed with depression, with a Bonferroni p value correction for multiple indicators. Accordingly, FMS negatively impacted both perceived physical and mental health status, although relatively more so for physical (p < 0.017). Women with FMS were more likely to have had reproductive health or sleep-related diagnoses, including premenstrual syndrome, dysmenorrhea, breast cysts, bladder cystitis, sleep apnea, restless leg syndrome, and abnormal leg movements (p < 0.0125). They were calculated to use less than half as many calories per week as control women (689 +/- 1293 vs. 1499 +/- 1584 kcal/week, p < 0.05) and had more sleep pattern difficulties (p < 0.0125), more negative changes in sexual function (greater odds for 5 of 10 indicators at p < 0.005), and lower alcohol use (odds ratio = 0.39, p < 0.05). CONCLUSIONS: Patients with FMS deserve careful assessment for reproductive conditions and sleep-related functional disorders. Besides more research into mechanisms underlying symptoms, intervention testing specifically to alleviate sleep problems, low physical activity levels, and sexual dysfunction should be paramount. PMID- 17125423 TI - The incidence of repeat uterine surgery following myomectomy. AB - OBJECTIVE: To determine the incidence of subsequent leiomyoma surgery following myomectomy in a population-based cohort. METHODS: All women, ages 20-75 years, enrolled in a large HMO in Washington State, identified by CPT and ICD-9 codes as having a myomectomy between January 1, 1993, and December 31, 2002, were eligible for the study. The follow-up period was 0.1-11 years. Automated enrollment and inpatient, outpatient, pathology, and procedure data were linked. Age-adjusted hazard ratios (HR) and 95% confidence intervals (CI) using Cox proportional hazard models, age-specific Kaplan Meier estimates of cumulative risk, and incidence of uterine surgery after myomectomy were calculated. RESULTS: Among 628 eligible women, 127 (21.8%) had a second surgery, 95 (74.8%) of which were hysterectomies. The cumulative incidence of a second surgery was 23.5% at 5 years and 30% at 7 years. During 2766.5 woman-years of follow-up, the annual incidence of a subsequent myoma surgery was 4.6%. Compared with women aged 40-44, women >or=50 had a 50% decreased risk for subsequent surgery (HR = 0.5, 95% CI 0.3 0.9). CONCLUSIONS: The incidence of subsequent surgery for leiomyomas following myomectomy is high, almost 5% per year. PMID- 17125424 TI - Women's satisfaction with an intravaginal Lactobacillus capsule for the treatment of bacterial vaginosis. AB - OBJECTIVE: To assess women's satisfaction with a vaginal capsule containing human derived, H(2)O(2)-producing Lactobacillus crispatus at completion of a randomized, placebo-controlled study for treatment of bacterial vaginosis (BV). METHODS: Women aged 14-35 years with BV were randomized to Lactobacillus or placebo intravaginal capsule twice daily for 3 days monthly for 3 months. Attitudes were assessed with a standardized scale at the final follow-up visit, and associations of satisfaction with clinical and gram stain cure of BV were determined. RESULTS: Four hundred twenty-four women were randomized at enrollment, and 232 (55%) provided data on satisfaction at the final visit. Overall, satisfaction with the capsule was high and was independent of subjects' race, age, and prior history of BV. Report of satisfaction with the capsule, belief that it contained healthy bacteria, and belief that its use improved vaginal health were directly related to clinical cure and to improved Nugent score. Except for women who had BV at the final follow-up visit, most subjects believed that the capsule contained healthy bacteria independent of any outcome reflecting cure. Willingness to use the capsule again was not significantly related to whether subjects experienced clinical cure or normalization of Nugent score at either visit. Adverse effects were rare and were largely related to a perceived difference in vaginal discharge. CONCLUSIONS: Satisfaction with an intravaginal capsule and positive beliefs about its use for BV treatment were prevalent, especially among women with a clinical and microbiological response to BV therapy. However, most women expressed willingness to use the product again regardless of clinical response. Acceptability of and interest in alternative BV treatments, such as L. crispatus intravaginal applications, are high. PMID- 17125425 TI - Changes in the prevalence of alcohol use during pregnancy among recent and at risk drinkers in the NLSY cohort. AB - PURPOSE: To support efforts to prevent fetal alcohol syndrome (FAS), population based data are needed on the prevalence of alcohol use at any time during gestation, particularly among women who were recent and at-risk drinkers. METHODS: We used National Longitudinal Survey of Labor Market Experiences in Youth (NLSY) files to estimate the prevalence of any drinking during pregnancy and to evaluate alcohol history risk factors among 6676 births reported by women with prepregnancy drinking data. Prevalence estimates were obtained for 2-year intervals for all 1982-1995 births and for subsets with prepregnancy recent and at-risk drinking. RESULTS: Among all births, drinking during pregnancy declined from 38.3% in 1982-1983 to 23.0% in 1994-1995 (p < 0.0001). Drinking during pregnancy also declined over time among recent and at-risk drinkers (p < 0.0001), but the 1994-1995 prevalences were still high (39.3% and 29.0%, respectively). Adjusted logistic models confirmed both the decrease in risk for the later birth years and the persistent heightened risk for births among recent and at-risk drinkers. CONCLUSIONS: In addition to ongoing universal prevention strategies that have helped reduce the prevalence of drinking during pregnancy, selective and indicated prevention approaches are needed to encourage abstinence during pregnancy among recent and at-risk drinkers. PMID- 17125426 TI - Clinical and personal intimate partner violence training experiences of U.S. medical students. AB - OBJECTIVE: To learn about U.S. medical students' attitudes, experiences, and practices regarding intimate partner violence (IPV). METHODS: In a sample reflective of all U.S. medical schools, we surveyed the class of 2003 in 16 U.S. medical schools at three different times in their training. RESULTS: A total of 2316 medical students responded, for a response rate of 80%. By senior year, although 91% of medical students reported receiving at least some training in discussing IPV, only one fifth reported extensive training. Although 73% of students entering wards thought IPV was highly important for physicians to discuss with patients, only 55% of students entering wards, decreasing to 35% of seniors, thought IPV would be highly relevant to their own practice. Only 55% of seniors reported talking with general medicine patients at least sometimes about IPV. Greater frequency of discussing IPV for seniors was associated with being a woman (60% vs. 50% for men, p = 0.006), self-designating as politically moderate or liberal (p = 0.0008), and thinking (on entering wards) that it was highly important for physicians to talk to patients about IPV (p = 0.0002). Perceived relevance of discussing domestic violence to intended practice was substantially higher among women, underrepresented minorities, those having a personal or family history of domestic violence, and those categorizing themselves as politically liberal or very liberal. Among seniors, the prevalence of reporting a personal history of IPV was 3% for women and 1% for men; 12% of women and 7% of men reported a family or personal IPV history. CONCLUSIONS: Despite national interest in IPV issues, efforts in U.S. medical schools to increase IPV screening and prevention have not achieved saturation. These gaps in IPV instruction in medical schools are a concern because studies have reported that physicians who receive IPV education training are significantly more likely to screen for it. PMID- 17125427 TI - Women's experiences with emergency contraception in an internal medicine practice. AB - BACKGROUND: Emergency contraceptive pills (ECPs) are effective for preventing unintended pregnancy. Whether patients in primary care settings receive physician counseling regarding ECPs has not been evaluated. METHODS: We conducted a cross sectional telephone survey of reproductive-age women who sought care at a university-based general internal medicine clinic regarding receipt of physician counseling about ECPs, knowledge and experiences with ECPs, and attitudes toward using ECPs. RESULTS: One hundred forty-nine women aged 18-45 completed the survey. Eighty percent of respondents (n = 119) were at risk for unintended pregnancy. Although all women in the sample had seen an internist in the previous 12 months, only 10% had received physician counseling about ECPs. There was little difference in the proportion of women who received counseling about ECPs comparing those who received care from an obstetrician/gynecologist and an internist with women who received care from an internist alone (13% vs. 8%, p = 0.529). Receipt of ECP counseling was not associated with the consistency of current contraceptive use. No women who were married or over the age of 40 were counseled about ECPs. The majority of participants (92%) had heard of ECPs, although most (54%) had learned about them through the media. Fifty-four percent of women would be likely to use ECPs to prevent unintended pregnancy. CONCLUSIONS: Only a fraction of women seeing internists for their primary care are receiving counseling about ECPs, irrespective of receiving care from an obstetrician/gynecologist. As primary care physicians, internists should determine risk for unintended pregnancy, assess patients' knowledge and attitudes toward ECPs, and provide counseling about this effective therapy. PMID- 17125428 TI - Managing persistent preventable threats to safer pregnancies and infant health in the United States: beyond silos and into integration, early intervention, and prevention. PMID- 17125431 TI - Pharmacokinetic bioequivalence of enfuvirtide using a needle-free device versus standard needle administration. AB - STUDY OBJECTIVES: To compare the relative bioavailability of enfuvirtide, a human immunodeficiency virus type 1 (HIV-1) fusion inhibitor, injected with the Biojector 2000 (B2000) needle-free device versus a 27-gauge half-inch needle syringe; and to assess safety, tolerability, and patient preference for the two devices. DESIGN: Open-label, randomized, two-period crossover bioequivalence evaluation. SETTING: Clinical research center. PATIENTS: Twenty-seven adults with HIV-1 viral loads below 1000 copies/ml. INTERVENTION: Each patient received enfuvirtide 90 mg subcutaneously with the B2000 and with the needle-syringe, with a 1-week washout between treatments. MEASUREMENTS AND MAIN RESULTS: Twenty-six and 27 patients were included in the bioequivalence and safety analyses, respectively. Plasma enfuvirtide concentrations were measured at baseline and at several intervals after each injection. The B2000:needle-syringe ratios of maximum concentration (C(max)), area under the concentration-time curve from time zero extrapolated to infinity (AUC(0-infinity)), and AUC from time zero to tau (dosing interval) (AUC(0-tau)) served as criteria for bioequivalence determination. The two drug delivery systems were considered bioequivalent if the 90% confidence intervals (CIs) for the ratios were within 0.8-1.25. Safety and tolerability were evaluated based on documentation of adverse events, graded laboratory toxicities, and local injection-site reactions. Patient surveys provided feedback on device preference. Ratios of C(max), AUC(0-infinity), and AUC(0-tau) were 0.95 (90% CI 0.84-1.09), 0.99 (90% CI 0.93-1.05), and 0.99 (90% CI 0.93-1.05), respectively. The frequency of injection-site reactions was low, and severity was generally mild for both devices. Survey results showed 18 patients (69%) had a positive overall impression of the B2000 and 14 (54%) felt safer injecting with this device. Overall, 17 patients (65%) preferred the B2000 over the needle-syringe. CONCLUSION: Bioavailability of enfuvirtide with the B2000 and needle-syringe was equivalent based on C(max), AUC(0-tau), and AUC(0 infinity). Safety profiles and injection-site reactions were comparable between the devices, but patients preferred the B2000. Delivery of enfuvirtide with the B2000 is a feasible alternative to standard needle administration and warrants further evaluation. PMID- 17125432 TI - Therapeutic levels of intravenous procainamide in neonates: a retrospective assessment. AB - STUDY OBJECTIVES: To evaluate dosing and pharmacokinetic parameters of intravenous continuous-infusion procainamide in neonates, and to identify dosage regimens and factors leading to therapeutic procainamide levels and minimal adverse events. DESIGN: Retrospective, observational study. SETTING: Pediatric hospital. PATIENTS: . Twenty-one patients (seven preterm, 14 full term) younger than 30 days who received continuous-infusion procainamide therapy for more than 15 hours or had two consecutive therapeutic procainamide levels obtained while receiving therapy between June 1, 2002, and December 31, 2005. MEASUREMENTS AND MAIN RESULTS: Data on demographics, dosing, drug levels, and adverse effects were collected. Doses that achieved therapeutic levels were documented, and procainamide clearance was calculated and evaluated with regard to renal function and gestational age in patients who were at steady state. Mean clearance and mean N-acetylprocainamide (NAPA):procainamide ratios were compared between preterm and term neonates. No patients experienced hemodynamic instability or other adverse effects due to procainamide. Procainamide was given as a mean +/- SD 9.6 +/- 1.5 mg/kg bolus in 20 of 21 patients before continuous infusion. The mean +/- SD dose at which two therapeutic levels were achieved was 37.56 +/- 13.52 microg/kg/minute. Procainamide clearance was 6.36 +/- 8.85 ml/kg/minute and correlated with creatinine clearance (r=0.78, p<0.00001) and age at day of sampling (r=0.49, p<0.00001). The NAPA:procainamide ratio at steady state was 0.84 +/- 0.53; two patients were determined to be fast acetylators (ratio > 1). Preterm infants had lower mean clearance rates (p<0.001) but higher NAPA:procainamide ratios (p<0.01) than those of term infants. Five patients experienced seven supratherapeutic levels while receiving therapy; four of these patients were preterm, and all had creatinine clearances less than 30 ml/minute/1.73 m(2). Three patients had four pairs of levels obtained after discontinuation of procainamide, and elimination rate constant and half-life were calculated. CONCLUSION: Procainamide can be safely used in neonates, with no short-term adverse effects. The dosage regimen for intravenous procainamide required to achieve therapeutic levels in neonates is similar to that of older infants and children. Doses may need to be reduced in premature infants and in those with renal dysfunction. PMID- 17125433 TI - Pharmacoeconomic evaluation of antimuscarinic agents for the treatment of overactive bladder. AB - STUDY OBJECTIVE: To compare the cost-effectiveness of various antimuscarinic agents for the treatment of overactive bladder (OAB). METHODS: A decision analysis model was developed and included clinical outcomes (i.e., therapy continued or discontinued, treatment success or failure, OAB-induced comorbidities) and costs for drugs and treatment of OAB-induced comorbidities (i.e., urinary tract infections, fractures, depression, and skin infections). Treatment success was defined as complete continence. A systematic MEDLINE literature search from January 1990-January 2006 identified English-language articles concerning the eight antimuscarinic drugs: darifenacin, solifenacin, trospium, immediate-release oxybutynin, extended-release oxybutynin, transdermal oxybutynin, immediate-release tolterodine, and extended-release tolterodine. Probabilities and cost data for these drugs were retrieved from the literature, and drug costs were based on 2005 average wholesale prices. The analysis was constructed from a payer's perspective. The time frame for the model was 3 months. RESULTS: Expected costs for each patient with OAB ranged from $3373 when treated with solifenacin to $3769 when treated with immediate-release oxybutynin. The average cost/patient with continued and successful treatment was lowest for solifenacin ($6863). Solifenacin dominated all other antimuscarinic agents because they were associated with high costs and low effectiveness. Success rates were the key parameters driving the sensitivity analysis. CONCLUSION: Among various antimuscarinic agents, solifenacin 5 mg had the lowest costs and highest effectiveness in the treatment of OAB. PMID- 17125434 TI - Pharmacotherapy considerations in advanced cardiac life support. AB - Cardiac arrest and sudden cardiac death remain major causes of mortality. Early intervention has been facilitated by emergency medical response systems and the development of training programs in basic life support and advanced cardiac life support (ACLS). Despite the implementation of these programs, the likelihood of a meaningful outcome in many life-threatening situations remains poor. Pharmacotherapy plays a role in the management of patients with cardiac arrest, with new guidelines for ACLS available in 2005 providing recommendations for the role of specific drug therapies. Epinephrine continues as a recommended means to facilitate defibrillation in patients with pulseless ventricular tachycardia or ventricular fibrillation; vasopressin is an alternative. Amiodarone is the primary antiarrhythmic drug that has been shown to be effective for facilitation of defibrillation in patients with pulseless ventricular tachycardia or fibrillation and is also used for the management of atrial fibrillation and hemodynamically stable ventricular tachycardia. Epinephrine and atropine are the primary agents used for the management of asystole and pulseless electrical activity. Treatment of electrolyte abnormalities, severe hypotension, pulmonary embolism, acute ischemic stroke, and toxicologic emergencies are important components of ACLS management. Selection of the appropriate drug, dose, and timing and route of administration are among the many challenges faced in this setting. Pharmacists who are properly educated and trained regarding the use of pharmacotherapy for patients requiring ACLS can help maximize the likelihood of positive patient outcomes. PMID- 17125435 TI - Factors influencing the magnitude and clinical significance of drug interactions between azole antifungals and select immunosuppressants. AB - The magnitude of drug interactions between azole antifungals and immunosuppressants is drug and patient specific and depends on the potency of the azole inhibitor involved, the resulting plasma concentrations of each drug, the drug formulation, and interpatient variability. Many factors contribute to variability in the magnitude and clinical significance of drug interactions between an immunosuppressant such as cyclosporine, tacrolimus, or sirolimus and an antifungal agent such as ketoconazole, fluconazole, itraconazole, voriconazole, or posaconazole. By bringing similarities and differences among these agents and their potential interactions to clinicians' attention, they can appreciate and apply these findings in a individualized patient approach rather than follow only the one-size-fits-all dosing recommendations suggested in many tertiary references. Differences in metabolism and in the inhibitory potency of cytochrome P450 3A4 and P-glycoprotein influence the onset, magnitude, and resolution of drug interactions and their potential effect on clinical outcomes. Important issues are the route of administration and the decision to preemptively adjust dosages versus intensive monitoring with subsequent dosage adjustments. We provide recommendations for the concomitant use of these agents, including suggestions regarding contraindicated combinations, those best avoided, and those requiring close monitoring of drug dosages and plasma concentrations. PMID- 17125436 TI - Entecavir: a new nucleoside analog for the treatment of chronic hepatitis B infection. AB - BACKGROUND: Chronic hepatitis B infection carries considerable risk for the development of cirrhosis and hepatocellular carcinoma. Treatment options are increasing but are limited to interferon alfa-2b, pegylated interferon alfa-2a, lamivudine, adefovir dipivoxil, and entecavir. Entecavir, a nucleoside analog, is the newest oral antiviral approved in the United States for treatment of chronic hepatitis B. OBJECTIVE: To review the available data for entecavir regarding its pharmacology, pharmacokinetics, safety, efficacy, and clinical use. METHODS: A MEDLINE, EMBASE, and Cochrane search of the English-language literature from January 1997-May 2006 was performed. Preapproval studies provided by the manufacturer and abstracts from recent scientific meetings on infectious disease and hepatology were also reviewed. RESULTS: Three phase III clinical trials representing more than 1600 subjects established the superior efficacy and equivalent safety of entecavir compared with lamivudine for treating patients who are hepatitis B early antigen (HBeAg) positive, HBeAg negative, or refractory to lamivudine. Entecavir resistance has not occurred in nucleoside-naive patients but may develop in those who already possess lamivudine resistance mutations. CONCLUSION: Trial results, along with previously published response rates for adefovir dipivoxil and interferon monotherapy, make entecavir the preferred first line treatment option for patients with chronic hepatitis B who are nucleoside naive, HBeAg positive or negative, and have compensated liver disease. Both entecavir and adefovir dipivoxil maintain activity against hepatitis B virus in patients with chronic hepatitis B who are refractory to lamivudine, and both agents are reasonable first-line treatment options. Longer trials involving nucleoside-naive, lamivudine-refractory patients are needed to determine entecavir's optimal treatment duration, long-term safety, and durability of response, including rate of resistance. PMID- 17125437 TI - Management of community-associated methicillin-resistant Staphylococcus aureus infections in children. AB - In recent years, community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has emerged as a pathogen in children without established risk factors, and its prevalence in the United States is increasing. Although many CA-MRSA infections are mild, primarily involving the skin and soft tissues, the organism can cause serious, invasive, and life-threatening infections. To provide a comprehensive review of the epidemiology, clinical features, therapy, and prevention of CA-MRSA infections in children, we performed MEDLINE (1966-January 2006) and Cochrane Library searches, and reviewed abstracts for relevance to S. aureus infections. Only articles pertaining to CA-MRSA infections in pediatrics were closely examined. As a genetically distinct pathogen, CA-MRSA is generally susceptible to multiple non-beta-lactam antimicrobials. The optimal treatment for CA-MRSA infections in pediatric patients has not been well studied. Common antibiotics used include clindamycin, trimethoprim-sulfamethoxazole, vancomycin, and rifampin. Rational empiric antimicrobial therapy for infections caused by S. aureus requires consideration of the possibility of methicillin resistance. The local prevalence and susceptibilities of CA-MRSA, severity of infection, and individual risk factors should be considered in selecting treatment. PMID- 17125438 TI - Rabbit antithymocyte globulin induction therapy in adult renal transplantation. AB - Rabbit antithymocyte globulin (rATG) and horse antithymocyte globulin (horse ATG) are the polyclonal antithymocyte agents available for use in solid organ transplantation in the United States. Horse ATG is indicated for induction immunosuppression and for treatment of acute rejection episodes after kidney transplantation; rATG is indicated for treatment of acute rejection only. However, rATG is commonly used in clinical practice as an induction immunosuppressive agent, instigating many questions regarding appropriate dosing, tolerability, safety, and efficacy. Available evidence supports the use of rATG as an induction agent in adult renal transplant recipients. The use of this product for induction therapy has been studied in conjunction with a full-dose, triple-therapy maintenance regimen (sequential quadruple immunosuppression) consisting of a calcineurin inhibitor, an antimetabolite, and corticosteroids. Rabbit ATG has a proven safety and efficacy profile both as treatment of acute rejection and as induction therapy in patients undergoing kidney transplantation. The most common adverse events associated with rATG are cytokine release syndrome, thrombocytopenia, and lymphopenia. Results of early studies showed an increased rate of cytomegalovirus disease associated with rATG treatment, but recent studies indicate that routine administration of modern antiviral prophylaxis can reduce this risk. Current practice with rATG is evolving to minimize lifelong exposure to calcineurin inhibitors and corticosteroids. PMID- 17125439 TI - Risk of serotonin syndrome with concomitant administration of linezolid and serotonin agonists. AB - Linezolid, an antimicrobial used to treat resistant gram-positive bacteria, can inhibit monoamine oxidase, an enzyme that metabolizes serotonin and other biogenic amines. Inhibition of this enzyme can predispose patients who are concomitantly taking serotonin agonists to serotonin syndrome. Because of the potential of linezolid to inhibit monoamine oxidase, premarketing studies were conducted with drugs such as selective serotonin reuptake inhibitors. No cases of serotonin syndrome were recorded. After linezolid was released to the United States market, several case reports of serotonin syndrome emerged. A literature search revealed 13 cases of serotonin syndrome occurring with the concomitant use of linezolid and drugs possessing serotonergic properties. To direct clinical management of this potential drug interaction, we reviewed reports of serotonin syndrome to determine relevant drug interactions with linezolid and serotonergic drugs and to characterize similarities and differences in the reported cases. Clinicians should obtain complete drug histories to identify patients at risk, strictly monitor drug therapy including concomitant drugs, and receive education about this potential drug interaction and the symptoms of serotonin syndrome. PMID- 17125440 TI - Complementary and alternative medicine use among individuals participating in research: implications for research and practice. AB - STUDY OBJECTIVES: To determine the frequency and type of complementary and alternative medicine (CAM) use among healthy volunteers participating in research, and to investigate the potential for interactions between commonly used CAM modalities and various drugs. DESIGN: Prospective evaluation. SETTING: University general clinical research center. SUBJECTS: Sixty healthy adults participating in an ongoing drug study. MEASUREMENTS AND MAIN RESULTS: The clinical study database was queried to determine the use and type of existing and newly started CAM throughout the study period. Baseline characteristics were compared between users and nonusers of CAM to identify differences between them. Potential CAM-drug interactions were classified based on curated databases and primary literature sources. Of the 60 subjects enrolled, 30 (50%) used CAM during the study. Of these, 26 (87%) were using CAM at study entry. Baseline CAM users were on average 7 years older than nonusers (p=0.03) and had high-density lipoprotein cholesterol concentrations 10 mg/dl higher than those of nonusers (p=0.04). The group using CAM had more women and nonsmokers than the other group. Several potential CAM-drug interactions were identified. CONCLUSION: Because of high rates of CAM use (50% of the subjects were using biologically based CAM) and the many potential CAM-drug interactions, CAM use should be rigorously addressed in clinical practice and research. Failure to capture this information may have clinical repercussions through pharmacokinetic and pharmacodynamic interference of clinical response and clinical trial results. Clinicians and researchers may be able to identify those most likely to use CAM by their baseline characteristics; this would help target those patients and research subjects for more thorough assessment and follow-up. PMID- 17125441 TI - Bezoar formation requiring endoscopic removal after intentional overdose of extended-release nifedipine. AB - A 61-year-old Caucasian woman was transported to the emergency department after intentionally ingesting several different prescription drugs. She had been found by her husband in an unconscious state with empty bottles of extended-release venlafaxine, extended-release nifedipine, sertraline, and atorvastatin. She was intubated in the emergency department and transferred to the intensive care unit. After 36 hours in the intensive care unit, she was stabilized and brought to a general medical ward. She later developed profound recurrent hypotension with systolic blood pressures ranging from 40-70 mm Hg and diastolic blood pressures of 0-40 mm Hg. She was readmitted to the intensive care unit, where a computed tomography scan revealed a mass in her stomach. A gastroenterology consultation was obtained, and an esophagogastroduodenoscopy (EGD) was performed, during which a large drug bezoar was discovered and removed. The drugs were identified as extended-release nifedipine with a few granules of extended-release venlafaxine. Unfortunately, the patient died 3 days after the EGD from multisystem organ failure related to the overdose. Clinicians who encounter drug overdoses should be aware of the possibility of drug bezoar formation and should consider endoscopic removal as a potential treatment option. PMID- 17125442 TI - Prolonged anticoagulation after discontinuation of argatroban and warfarin therapy in an obese patient with heparin-induced thrombocytopenia. AB - A 32-year-old, morbidly obese African-American woman developed bilateral pulmonary emboli 12 days after undergoing Roux-en-Y gastric bypass surgery. Three days later, after receiving heparin and warfarin, she developed heparin-induced thrombocytopenia type II (HIT-II). An argatroban 1.5-microg/kg/minute infusion was administered for approximately 2.5 days. The patient also received four doses of warfarin, totaling 37.5 mg. The argatroban infusion was discontinued early on hospital day 6, at which time the patient's international normalized ratio (INR) was 4.36 and activated partial thromboplastin time (aPTT) 85.9 seconds. Her INR and aPTT values continued to rise after the argatroban was discontinued and peaked 3 days later at 5.28 and 123.6 seconds, respectively. At this time her platelet count had improved from 139 x 10(3)/mm(3) to 543 x 10(3)/mm(3). No additional warfarin was administered before discharge. On hospital day 11, the patient was discharged home with an INR of 4.12 and an aPTT of 67.1 seconds. Her aPTT and INR values remained elevated for 19 days after receiving her last dose of warfarin and for 20 days after argatroban discontinuation. She experienced no bleeding complications from these supratherapeutic coagulation parameters. She resumed treatment with warfarin as an outpatient and completed a 6-month course of anticoagulation without further incident. Clinicians should be aware that coagulation parameters may remain elevated longer than expected after argatroban discontinuation in certain patients taking concomitant warfarin. Patients with liver dysfunction and obesity appear most likely to be affected. PMID- 17125443 TI - Recurrent anaphylaxis associated with gonadotropin-releasing hormone analogs: case report and review of the literature. AB - An 8-year-old girl with idiopathic central precocious puberty experienced multiple episodes of anaphylaxis after receiving a goserelin acetate implant. She was hospitalized and treated with epinephrine, antihistamine, and corticosteroids. The goserelin implant was surgically excised; however, anaphylactic symptoms continued for 4 days after excision. Less severe systemic symptoms recurred 6 weeks after removal; these were possibly due to leakage of the depot drug into subcutaneous tissues. It was noted that 3 years earlier, the patient had developed a similar, milder systemic allergic reaction to leuprolide acetate that required treatment with oral prednisone and antihistamines. Intradermal testing yielded positive results for leuprolide. Gonadotropin releasing hormone (GnRH) analogs, including leuprolide acetate and goserelin acetate, are commonly prescribed for patients with prostatic carcinoma, endometriosis, and precocious puberty. A literature review identified a single case report of a systemic hypersensitivity reaction involving goserelin acetate and several reports of systemic hypersensitivity reactions associated with leuprolide acetate. We found no reports of systemic reactions to GnRH analogs in pediatric patients. Clinicians should be aware of the potential association of GnRH analogs with systemic reactions. They should also recognize that recurrent anaphylaxis may occur due to the long half-life of these therapeutic agents in tissue. PMID- 17125444 TI - Future clinical pharmacy practitioners should be board-certified specialists. PMID- 17125447 TI - Challenges associated with metabolic syndrome. AB - Approximately 2500 Americans die from cardiovascular disease (CVD) each day. Each year, CVD claims more lives than the next four leading causes of death combined. Direct and indirect costs of CVD are estimated to be Dollars 403.1 billion in 2006. Despite advancements in conventional therapy, the residual risk of CVD continues to rise. One component of the cardiometabolic risk is metabolic syndrome. Metabolic syndrome is a constellation of interrelated risk factors of metabolic origin including abdominal obesity, atherogenic dyslipidemia, elevated blood pressure, elevated plasma glucose level, and prothrombotic and proinflammatory states that promote atherosclerotic CVD and increase the risk of type 2 diabetes mellitus. Approximately 47 million residents of the United States have metabolic syndrome. Abdominal obesity and insulin resistance appear to be its predominant underlying risk factors. Abdominal adiposity is considered high risk fat, and it is associated with insulin resistance, hyperglycemia, dyslipidemia, hypertension, and prothrombotic and/or proinflammatory states. Despite notable advances in cardiovascular risk management, the prevalence of cardiovascular events and type 2 diabetes remains high. First-line therapy for individuals with metabolic syndrome should be directed to the major CVD risk factors, namely, elevated low-density lipoprotein cholesterol levels, hypertension, and diabetes, and it should emphasize lifestyle modification. Until additional research better defines the most appropriate therapies, conventional cardiovascular risk factors, such as lipid levels, blood pressure, and diabetes, should be managed in individuals with metabolic syndrome according to nationally accepted clinical guidelines. PMID- 17125448 TI - The endocannabinoid system. AB - The endocannabinoid system is a complex physiologic system. One of the most important discoveries related to the endocannabinoid system is that cannabinoid-1 receptors are present throughout the body and that they are linked to obesity and cardiometabolic risk. Adipose tissue was historically thought to be an inert, passive storage vehicle. However, recent findings prove that adipose tissue is a complex endocrine organ that releases adipokines, which influence cardiometabolic risk factors. Elevated endocannabinoid system activity is associated with obesity and coupled with excessive food intake and fat accumulation. Strong evidence suggests that the endocannabinoid system affects the surrogate cardiometabolic end points that greatly influence morbidity and mortality. PMID- 17125449 TI - Metabolic syndrome treatment strategies. AB - The primary goal of managing patients with metabolic syndrome is to decrease their cardiovascular risk by individualizing treatment strategies according to the patient's specific risk factors. Treatment options for patients with metabolic syndrome include lifestyle modification and drug therapy. Lifestyle modification can be summarized as dietary changes, exercise, and smoking cessation. Drug therapy indicated for cardiometabolic risk reduction includes antihypertensives, insulin sensitizers, and cholesterol-lowering agents. In addition, two drugs-sibutramine and rimonabant-have been evaluated and produced promising outcomes in the overall management of high-risk patients with metabolic syndrome. PMID- 17125450 TI - Role of the pharmacist. AB - The prevalence of cardiovascular risk factors continues to rise, with considerable emphasis on the upward trends in obesity and diabetes mellitus, which have increased by 46% and 60%, respectively, in the past 10 years. The alarming increase in the prevalence of diabetes and obesity has resulted in the development of new and innovative drug therapies to assist in managing cardiometabolic risk factors. This advent creates many opportunities for pharmacists to evaluate patients' drug regimens and influence lifestyle modification. Drug therapy management programs allow pharmacists to take active roles in assessing drug regimens and to intervene as appropriate. No matter what their practice environment, pharmacists have numerous opportunities to recognize and recommend treatment for cardiometabolic risk factors and to increase patient compliance by educating patients and health care practitioners. PMID- 17125452 TI - Molecular interactions of acute phase serum amyloid A: possible involvement in carcinogenesis. AB - Acute phase serum amyloid A (A-SAA) is a well-known marker of inflammation. The present review summarizes data on the regulation of A-SAA expression, signaling pathways which it is involved in, its effects, and possible influences on progression of malignant tumors. PMID- 17125453 TI - Quantitative proteomics and its applications for systems biology. AB - Here we discuss the current state of research in the rapidly growing field of quantitative proteomics and its applications to systems biology. Quantitative proteomics can be successfully used for characterizing alterations in protein abundance, finding novel protein-protein and protein-peptide interactions, investigating formation of large macromolecular complexes, and elucidating temporal changes in organellar protein composition and phosphorylation in signal transduction cascades. Further, quantitative proteomics can directly compare activation of entire signaling networks in response to individual stimuli and discover critical differences in their circuits that account for alterations of cell response. Maturation of proteomic bioinformatics applications and continuous improvements in proteomics and related genomics and transcriptomics technologies now allows us to investigate cellular mechanisms at the integrative system level. PMID- 17125454 TI - Glucocorticoid resistance. AB - Glucocorticoids contribute fundamentally to the maintenance of basal and stress related homeostasis in all higher organisms. The major roles of these steroids in physiology are amply matched by their remarkable contributions to pathology. Glucocorticoid resistance is a rare familial, or sporadic condition characterized by partial end-organ insensitivity to glucocorticoids. The molecular basis of glucocorticoid resistance in several families and sporadic cases has been ascribed to mutations in the human glucocorticoid receptor alpha (hGRalpha) gene, which impair the ability of the receptor to transduce the glucocorticoid signal. Glucocorticoids are crucial for life, and therefore complete glucocorticoid resistance is uncommon. The purpose of this review is to discuss the many structural and functional features of the glucocorticoid receptor and also to evaluate the main clinical and laboratory characteristics of cortisol resistance. PMID- 17125455 TI - Modulation of hippocampal neuron survival by thrombin and factor Xa. AB - Effects of thrombin, factor Xa (FXa), and protease-activated receptor 1 and 2 agonist peptides (PAR1-AP and PAR2-AP) on survival and intracellular Ca2+ homeostasis in hippocampal neuron cultures treated with cytotoxic doses of glutamate were investigated. It is shown that at low concentrations ( KClO4 > K2SO4 > KCl. For alcohols, this order was found to be as follows: butanol > propanol > ethanol > methanol. PMID- 17125459 TI - Silybin reduces lipid peroxidation of rat hepatocyte membrane caused by cyclosporin A. AB - An effect of cyclosporin A on lipid peroxidation in isolated rat hepatocytes was tested. A significant increase in lipid peroxidation marker (the concentration of lipofuscin-like pigments) was observed in samples incubated with cyclosporin A in comparison with the control. When hepatoprotective flavonoid silybin was added, the production of lipofuscin-like pigments decreased significantly. This result indicates a potential positive role of silybin in lowering of cyclosporin A side effects associated with the production of reactive oxygen species and plasma membrane damage. PMID- 17125460 TI - Microcystin-induced 8-hydroxydeoxyguanosine in DNA and its reduction by melatonin, vitamin C, and vitamin E in mice. AB - Microcystin LR (MC-LR), a liver-specific toxin synthesized by Microcystis aeruginosa, was investigated. MC-LR initiated reactive oxygen species formation followed by damaging DNA and some other cellular components. We investigated the ability of MC-LR to induce oxidative DNA damage by examining the formation of 8 hydroxydeoxyguanosine (8-OH-dG) using HPLC with electrochemical detection. Melatonin, vitamin C (ascorbate), and vitamin E (as Trolox), all of which are free radical scavengers, markedly inhibited the formation of 8-OH-dG in a concentration-dependent manner. The concentration that reduced DNA damage by 50% (IC(50)) was 0.55, 31.4, and 36.8 microM for melatonin, ascorbate, and Trolox, respectively. The results show that melatonin is 60- and 70-fold more effective than vitamin C or vitamin E, respectively, in reducing oxidative DNA damage. These findings are consistent with the conclusion that melatonin's highly protective effect against microcystin toxicity relates, at least in part, to its direct hydroxyl radical scavenging ability. PMID- 17125461 TI - Death of stoma guard cells in leaf epidermis under disturbance of energy provision. AB - Cyanide is an apoptosis inducer in stoma guard cells from pea leaf epidermis. Unlike CN-, the uncoupler of oxidative and photosynthetic phosphorylation carbonyl cyanide m-chlorophenylhydrazone (CCCP), the combination of CCCP, 3-(3 ,4 -dichlorophenyl)-1,1-dimethylurea (DCMU), benzylhydroxamate (BH), myxothiazol, antimycin A, and a glycolysis inhibitor 2-deoxyglucose (DG) did not induce destruction of guard cell nuclei for 20 h of incubation of epidermal peels in the light. DCMU prevented the effect of CN- as a programmed cell death (PCD) inducer. CCCP, the combination of DCMU and CCCP, or the combination of DCMU, CCCP, BH, myxothiazol, antimycin A, and DG supplemented by CN- caused destruction of cell nuclei; the number of the cells lacking nuclei in this case was higher than with CN- alone. DG and CCCP caused cell destruction after longer incubation of the isolated epidermis - after 2 days and to a greater degree after 4 days. The effect of DG and CCCP was reduced by illumination. Cell destruction during long term incubation was prevented by the combination of DG and CCCP. From data of electron microscopy, DCMU and dinitrophenyl ester of iodonitrothymol (DNP-INT) prevented apoptotic changes of the nuclear ultrastructure induced by CN-. The suppression of the destruction of the guard cell nuclei under combined action of DG and CCCP was apparently caused by switching of cell death from PCD to necrosis. Thus, the type of cell death - via apoptosis or necrosis - is controlled by the level of energy provision. PMID- 17125462 TI - Regulation of cytochrome C peroxidase activity by nitric oxide and laser irradiation. AB - Apoptosis can be induced by activation of so-called "death receptors" (extrinsic pathway) or multiple apoptotic factors (intrinsic pathway), which leads to release of cytochrome c from mitochondria. This event is considered to be a point of no return in apoptosis. One of the most important events in the development of apoptosis is the enhancement of cytochrome c peroxidase activity upon its interaction with cardiolipin, which modifies the active center of cytochrome c. In the present work, we have investigated the effects of nitric oxide on the cytochrome c peroxidase activity when cytochrome c is bound to cardiolipin or sodium dodecyl sulfate. We have observed that cytochrome c peroxidase activity, distinctly increased due to the presence of anionic lipids, is completely suppressed by nitric oxide. At the same time, nitrosyl complexes of cytochrome c, produced in the interaction with nitric oxide, demonstrated sensitivity to laser irradiation (441 nm) and were photolyzed during irradiation. This decomposition led to partial restoration of cytochrome c peroxidase activity. Finally, we conclude that nitric oxide and laser irradiation may serve as effective instruments for regulating the peroxidase activity of cytochrome c, and, probably, apoptosis. PMID- 17125463 TI - Red fluorescent protein DsRed: parametrization of its chromophore as an amino acid residue for computer modeling in the OPLS-AA force field. AB - Topology of the neutral form of the DsRed fluorescent protein chromophore as a residue of [(4-cis)-2-[(1-cis)-4-amino-4-oxobutanimidoyl]-4-(4 hydroxybenzylidene)-5-oxo-4,5-dihydro-1H-imidazol-1-yl]acetic acid was calculated with OPLS-AA force field. Use of this topology and molecular dynamics simulation allows calculating the parameters of proteins that contain such residue in their polypeptide chains. The chromophore parameters were obtained by ab initio (RHF/6 31G**) quantum chemical calculations applying density functional theory (B3LYP). Using this chromophore, we have calculated the molecular dynamics trajectory of tetrameric fluorescent protein DsRed in solution at 300 K (4 nsec). Correctness of the chromophore parametrization was revealed by comparison of quantitative characteristics of the chromophore structure obtained from the molecular dynamic simulations of DsRed protein with the quantitative characteristics of the chromophore based on the crystallographic X-ray data of fluorescent protein DsRed (PDB ID: 1ZGO, 1G7K, and 1GGX), and also with the quantitative characteristics of the chromophore obtained by quantum chemical calculations. Inclusion of the neutral form of DsRed protein chromophore topology into the OPLS-AA force field yielded the extended force field OPLS-AA/DsRed. This force field can be used for molecular dynamics calculations of proteins containing the DsRed chromophore. The parameter set presented in this study can be applied for similar extension in any other force fields. PMID- 17125464 TI - Downregulation of genes encoding for subunits of adaptor complex-3 in cervical carcinomas. AB - We explored the expression of four genes encoding for subunits of AP-3 in cervical tumors and cancer cell lines. Using RT-PCR we demonstrated more than twofold decrease in the levels of mRNA of AP3D1, AP3B1, AP3M1, and AP3S1 in 32, 28, 23, and 26% tumors in comparison with normal tissues of uterine cervix, respectively. The level of mRNA of at least one subunit was decreased in 28 out of 47 (60%) of tumors and in four out of five cancer cell lines in comparison to tissues adjacent to tumors. The suppression of expression of any of the subunits was revealed in 15 out of 28 cases (54%). The expression of two and more subunits was decreased simultaneously in different combinations in 13 cases (46%). This fact testifies to the lack of a common mechanism of downregulation of four subunits in tumors. There is a tendency to more frequent suppression of AP-3A expression in tumors associated with lymphatic node metastases as compared with tumors without metastases (P = 0.034). Thus, here we demonstrate for the first time the decrease in expression of genes encoding for AP-3A subunits in tumors. PMID- 17125465 TI - Properties of yeast Saccharomyces cerevisiae plasma membrane dicarboxylate transporter. AB - Transport of succinate into Saccharomyces cerevisiae cells was determined using the endogenous coupled mitochondrial succinate oxidase system. The dependence of succinate oxidation rate on the substrate concentration was a curve with saturation. At neutral pH the K(m) value of the mitochondrial "succinate oxidase" was fivefold less than that of the cellular "succinate oxidase". O-Palmitoyl-L malate, not penetrating across the plasma membrane, completely inhibited cell respiration in the presence of succinate but not glucose or pyruvate. The linear inhibition in Dixon plots indicates that the rate of succinate oxidation is limited by its transport across the plasmalemma. O-Palmitoyl-L-malate and L malate were competitive inhibitors (the K(i) values were 6.6 +/- 1.3 microM and 17.5 +/- 1.1 mM, respectively). The rate of succinate transport was also competitively inhibited by the malonate derivative 2-undecyl malonate (K(i) = 7.8 +/- 1.2 microM) but not phosphate. Succinate transport across the plasma membrane of S. cerevisiae is not coupled with proton transport, but sodium ions are necessary. The plasma membrane of S. cerevisiae is established to have a carrier catalyzing the transport of dicarboxylates (succinate and possibly L-malate and malonate). PMID- 17125466 TI - AT1 receptor blockade prevents the decrease in conduit artery flow-mediated dilatation during NOS inhibition in humans. AB - Whether AT(1) (angiotenin II type 1) receptor blockade can prevent the decrease in conduit artery FMD (flow-mediated dilatation) during NOS (nitric oxide synthase) inhibition by alternative endothelial pathways has not been explored previously in humans. In 12 healthy subjects, we measured radial artery diameter (echotracking) and flow (Doppler) during FMD induced by sustained reactive hyperaemia during a control period and following NOS inhibition [1.5 mg.min( 1).l(-1) L-NMMA (N(G)-monomethyl-L-arginine)], after a single oral administration of telmisartan (80 mg) or placebo, using a randomized double-blind cross-over design. In six volunteers, we also assessed the roles of prostacyclin and EDHF (endothelium-derived hyperpolarizing factor) during radial FMD after AT(1) receptor blockade by oral administration of aspirin (500 mg) alone, aspirin+L NMMA or aspirin+L-NMMA+fluconazole (a cytochrome epoxygenases inhibitor; 0.37 mg.min(-1).l(-1)). Telmisartan did not affect radial artery FMD in the control period (10.9+/-0.6% with placebo compared with 9.9+/-0.7% with telmisartan), but prevented its decrease after L-NMMA (9.3+/-0.8% with placebo compared with 12.6+/ 1.2% with telmisartan; P<0.05) with no modification in baseline parameters, hyperaemia and radial artery endothelium-independent dilatation to sodium nitroprusside. Moreover, in telmisartan-treated subjects, radial artery FMD, compared with control (9.0+/-1.0%), was not modified by aspirin alone (9.4+/ 0.7%) or associated with L-NMMA (9.5+/-0.5%), but was reduced by the combination of aspirin, L-NMMA and fluconazole (7.5+/-0.6%; P<0.05). These results demonstrate that AT(1) receptor blockade prevents the decrease in conduit artery FMD during NOS inhibition in humans, suggesting the development of a compensatory endothelial mechanism. This mechanism appears to be independent of prostacyclin and could possibly be related to an EDHF release. PMID- 17125469 TI - Prostate-specific antigen kinetics and the diagnosis and management of prostate cancer. PMID- 17125467 TI - Aurora-A kinase interacting protein 1 (AURKAIP1) promotes Aurora-A degradation through an alternative ubiquitin-independent pathway. AB - Mitotic Aurora-A is an oncogene, which undergoes a cell-cycle-dependent regulation of both its synthesis and degradation. Overexpression of Aurora-A leads to aneuploidy and cellular transformation in cultured cells. It has been shown that the cell-cycle-dependent turnover of Aurora-A is mediated by Cdh1 (CDC20 homologue 1) through the anaphase-promoting complex/cyclosome (APC/C) ubiquitin-proteasome pathway. We have described previously the identification of an Aurora-A kinase interacting protein, AURKAIP1 (formerly described as AIP), which is also involved in the destabilization of Aurora-A through the proteasome dependent degradation pathway. In an attempt to investigate the mechanism of AURKAIP1-mediated Aurora-A degradation, we report here that AURKAIP1 targets Aurora-A for degradation in a proteasome-dependent but Ub (ubiquitin)-independent manner. AURKAIP1 inhibits polyubiquitination of Aurora-A. A non-interactive AURKAIP1 mutant that cannot destabilize Aurora-A restores ubiquitination of Aurora-A. An A-box mutant of Aurora-A, which cannot be targeted for proteasome dependent degradation by Cdh1, can still be degraded by AURKAIP1. Inhibition of cellular ubiquitination either by expression of dominant negative Ub mutants or by studies in ts-20 (temperature sensitive-20) CHO (Chinese-hamster ovary) cell line lacking the E1 Ub activating enzyme at the restrictive temperature, cannot abolish AURKAIP1-mediated degradation of Aurora-A. AURKAIP1 specifically decreases the stability of Aurora-A in ts-20 CHO cells at the restrictive temperature, while cyclinB1 and p21 are not affected. This demonstrates that there exists an Ub-independent alternative pathway for Aurora-A degradation and AURKAIP1 promotes Aurora-A degradation through this Ub-independent yet proteasome dependent pathway. PMID- 17125470 TI - Current opinion amongst radiologists and urologists in the UK on percutaneous nephrostomy and ureteric stent insertion for acute renal unobstruction: Results of a postal survey. PMID- 17125471 TI - The need for supervised training in urology outpatients: A case for restructuring. PMID- 17125472 TI - Renal 'perivascular epitheloid cell-omas'. PMID- 17125473 TI - The role of botulinum toxin in benign prostatic hyperplasia. PMID- 17125474 TI - Nephroureterectomy for treating upper urinary tract transitional cell carcinoma: Time to change the treatment paradigm? AB - OBJECTIVE: To evaluate factors associated with disease recurrence and survival in patients undergoing nephroureterectomy for upper urinary tract transitional cell carcinoma (UUT-TCC) in one centre over an 18-year period. PATIENTS AND METHODS: The records of patients who had a nephroureterectomy for UUT-TCC at our institution from 1986 to 2004 were reviewed for clinical, pathological and treatment period data. Cox's proportional hazards regression model was used to test the statistical significance of several potential prognostic factors for recurrence and survival. RESULTS The median overall duration of follow-up was 2.5 years for 184 patients. Significant prognosticators for disease-specific survival (DSS) by univariate analysis were tumour stage (P < 0.01), tumour grade (P < 0.01), node-positive disease (P < 0.01), multifocality (P = 0.03), previous cystectomy (P < 0.01) and synchronous bilateral UUT-TCC (P = 0.02). On multivariate analysis, only tumour stage (P = 0.03) and grade (P = 0.01) correlated with DSS. The median recurrence-free survival duration was 2.4 years. In 44 patients, the disease recurred outside the bladder; 15 (8.2%) had local recurrence, 20 (10.9%) distant metastasis, and nine (4.9%) both local and distant recurrence. Bladder tumours occurred in 40 (26.1%) patients with no previous cystectomy. The evaluation of treatment outcome during three periods of the study showed no significant effect on DSS. CONCLUSION: Tumour stage and grade correlated with DSS in this cohort, with no improvement in outcome over the 18 year period assessed. Patients with high-stage and high-grade disease continue to fare poorly, suggesting a need for changing the treatment protocol. Judiciously applying a multimodal approach to the management of high-risk patients by incorporating neoadjuvant chemotherapy and surgical resection might provide, for the first time, the opportunity to improve patient outcome. PMID- 17125475 TI - Bladder tumour development after urothelial carcinoma of the upper urinary tract is related to primary tumour location. AB - OBJECTIVE: To better define the predictors of bladder tumour development in patients operated for upper urinary tract urothelial cancer (UT-UC). PATIENTS AND METHODS: Surgical specimens from 191 consecutive patients with no history of bladder cancer and operated for UT-UC were chosen for analysis. Bladder tumour development was assessed in relation to UT-UC location, tumour multiplicity, stage and grade, margin status, mode of operation, age and gender. RESULTS: Overall, 51 of 191 (27%) patients developed subsequent bladder tumours, including 25 of 123 (20%) with pelvic, 19 of 47 (40%) with ureteric and seven of 21 (33%) with multifocal tumours (P = 0.04 for all subgroups; P = 0.01 for pelvic vs ureteric). There was no influence of the other variables. The median (mean, range) time to recurrence was 12 (18, 3-64) months. In a multivariate analysis, ureteric tumour location was an independent predictor (P = 0.02; risk ratio, RR, 2.0, 95% confidence interval, CI, 1.1-3.7). After excluding 68 patients with systemic disease progression, bladder tumour development was noted in 39 of 123 (32%), including 18 of 76 (24%) with pelvic, 16 of 34 (47%) with ureteric and five of 13 with multifocal tumours (P = 0.06 for all subgroups; P = 0.02 for pelvic vs ureteric). In a multivariate analysis, ureteric location (P = 0.03; RR 2.1, 95% CI 1.1-4.2) and high tumour grade (P = 0.04; RR 2.2, 95% CI 1.03-4.7) were independent predictors of subsequent bladder tumour development. CONCLUSION: The risk of developing a bladder tumour after surgery for UT-UC is significantly related to ureteric tumour location and high tumour grade. Clinical trials to evaluate a possible reduction of bladder cancer risk by intraoperative ureteric ligation and/or peri-operative topical intravesical chemotherapy instillation are justified. PMID- 17125476 TI - Visually directed high-intensity focused ultrasound for organ-confined prostate cancer: A proposed standard for the conduct of therapy. AB - OBJECTIVE: To propose a standard for the conduct of visually directed transrectal high-intensity focused ultrasound (HIFU) and to offer a formal description of the changes observed on B-mode ultrasonography (US) during this procedure. We describe our early experience of using two different treatment methods; algorithm based HIFU and visually directed HIFU for the treatment of organ-confined prostate cancer. PATIENTS AND METHODS: Between November 2004 and October 2005, 34 men were treated using the Sonablate-500 (Focus Surgery, Indianapolis, IN, USA) as primary therapy for T1 or T2 prostate cancer. None had had previous hormone therapy and all had > or = 3-month PSA nadirs recorded at the follow-up. Nine men were treated using an algorithm-based protocol (group 1) and 25 using visually directed therapy (group 2). The conduct of visually directed treatment was described and changes seen using B-mode US were categorized using three 'Uchida' grades. RESULTS The mean PSA nadir achieved in group 2 was 0.15 ng/mL, vs 1.51 ng/mL in group 1 (P < 0.005). In group 2, 21 of 25 men achieved PSA nadirs of < or = 0.2 ng/mL 3 months after treatment. Seven men achieved undetectable PSA values. The occurrence rate of treatment-related toxicity was similar in both groups. CONCLUSION: Visually directed, transrectal HIFU enables clinically important and statistically significantly lower PSA nadirs to be achieved than algorithm-based HIFU. This is the first reported experience of visually directed HIFU for the treatment of organ-confined prostate cancer. We think that this is the first attempt to standardize the conduct of therapy; such standardization facilitates teaching it, and makes it possible to derive quality standards. The standardization of the conduct of therapy is a key step in the process of health technology assessment. PMID- 17125477 TI - Short-term outcome after high-intensity focused ultrasound in the treatment of patients with high-risk prostate cancer. AB - OBJECTIVE: To assess the short-term outcome in patients with high-risk prostate cancer treated by transrectal high-intensity focused ultrasound (HIFU). PATIENTS AND METHODS: From April 2003 to November 2004, 30 patients with high-risk prostate cancer were enrolled in this prospective study; all had transurethral resection of the prostate before transrectal HIFU treatment, using the Ablatherm device (EDAP, Lyon, France) during the same session, associated with hormonal therapy with luteinizing hormone-releasing hormone analogues. After the procedure, all the patients were evaluated every 3 months by physical examination, prostate-specific antigen (PSA) assay and a continence questionnaire. The follow-up schedule also included a transperineal prostate biopsy 6 months after the treatment. All the patients had a minimum follow-up of 12 months. RESULTS: The HIFU treatment took a median (interquartile range, IQR) of 140 (100-160) min. No complications were reported during treatment. The mean (IQR) hospitalization was 2.2 (1-4) days, and the suprapubic drainage tube was removed after 12 (7-18) days. The complications after treatment were: urinary tract infections in five patients (16%), stenosis of the intraprostatic and membranous urethra in three (10%), and secondary infravesical obstruction in four (13%). At 12 months after the procedure, 28 patients (93%) were continent. Seven of the 30 men (23%) had a positive prostate biopsy. At the 1-year follow-up only three of the 30 patients with high-risk prostate cancer had a PSA level of >0.3 ng/mL. CONCLUSIONS: HIFU is a modern, minimally invasive therapy for prostate cancer, often used in selected patients with localized disease. The present results show that HIFU was also feasible in patients with high-risk prostate cancer. The low complication rates and favourable functional outcome support the planning of further larger studies in such patients. The oncological efficacy of HIFU should be assessed in further studies with a longer follow-up. PMID- 17125478 TI - Impact of surgical margin status on long-term cancer control after radical prostatectomy. AB - OBJECTIVE: To determine whether previously described technical modifications that significantly decreased the positive surgical margin (PSM) rate have translated into improved long-term cancer control, as SM status is generally recognized as an independent risk factor for biochemical recurrence (BR) after radical retropubic prostatectomy (RRP), and is the only factor that can be modified by surgical technique. PATIENTS AND METHODS: Between March 1994 and December 2004, 996 consecutive patients had RRP as the sole treatment for clinically localized prostate cancer. The surgery was done by one surgeon (C.B.B.) and the data were prospectively reviewed. RESULTS: The overall PSM rate was 8.8%; the PSM rate by pathological stage was 1.7%, 24.2% and 27.1% for men with pT2, pT3a and pT3b disease, respectively (P < 0.001). In all, 968 of 996 (97.2%) patients were available for the follow-up (mean 6.4 years); 69 of 883 (7.8%) with negative SMs (NSMs) developed BR, vs 29 of 85 (34%) with PSMs (P < 0.001). The actuarial 5- and 10-year biochemical disease-free survival was 92.1% and 89.6%, and 70.6% and 59.9%, for patients with NSM and PSM, respectively (P < 0.001). On multivariate analysis, PSM, pathological stage and Gleason grade were the strongest predictors of BR (P < 0.001). The preoperative prostate-specific antigen level, and clinical stage T1c and T2a disease were not associated with recurrence. The hazard ratio (95% confidence interval) for BR in patients with PSMs was 3.27 (2.1-5.1). CONCLUSIONS: RRP including the previously described surgical modifications not only decreased the PSM rate but also resulted in excellent long-term cancer control. The importance of meticulous surgical technique in RRP cannot be overemphasised. PMID- 17125479 TI - Risk and presenting features of prostate cancer amongst African-Caribbean, South Asian and European men in North-east London. AB - OBJECTIVE: To determine whether there are ethnic differences in the incidence and presenting features of all patients with prostate cancer presenting in North-east London, UK. PATIENTS AND METHODS: All newly diagnosed men with prostate cancer between 1999 and 2000 who were resident in the East London and City Health Authority were identified from various sources. Key clinical features were extracted from hospital records. The age-adjusted incidence rates for European, South Asian and African-Caribbean patients were calculated using census denominator data. RESULTS: For men aged >50 years the annual age-adjusted incidence rates (95% confidence interval) were 213 (186-240), 647 (504-789) and 199 (85-310) for the European, African-Caribbean and South Asian patients, respectively. African-Caribbean men had a three times greater risk (risk ratio 3.07, 2.40-3.93, P < 0.001) than European men. South Asian men had a lower risk of prostate cancer but this could have been compatible with chance. There was no evidence of marked ethnic differences for prostate-specific antigen levels, clinical staging and Gleason scores. CONCLUSION: The greater risk of prostate cancer for African-Caribbean men in South-east England is consistent with data from the USA and the Caribbean. Future work needs to determine whether this risk differs according to country of origin, and which genetic and/or environmental risk factors might be important in explaining these observations. PMID- 17125480 TI - Cisplatin, methotrexate and bleomycin for treating advanced penile carcinoma. AB - OBJECTIVE: To retrospectively evaluate the efficacy and toxicity of chemotherapy with cisplatinum, methotrexate and bleomycin (CMB) in the adjuvant and palliative setting, and its effect on survival in patients with locally advanced or metastatic penile carcinoma, which carries a very poor prognosis. PATIENTS AND METHODS: Thirteen patients (mean age 54 years) with radically resected local and nodal disease (eight), metastatic disease (two) or metastatic recurrence (three) had chemotherapy with CMB between February 1996 and July 2003. In all, 45 treatment courses were given, with a mean (range) of 3.5 (2-6) courses per patient. RESULTS: Three of the eight patients with adjuvant treatment showed no evidence of disease after a mean (range) of 54 (41-76) months, while four in this group died from disease progression after a mean of 11 (5-20) months, and one died from treatment-related toxicity. All five patients with metastatic disease died from disease progression after three had shown temporary signs of regression. CONCLUSIONS: Chemotherapy with CMB had little effect on metastatic penile cancer and responses were transient. However, patients with minimal disease after radical local and lymphatic resection seemed to benefit from adjuvant therapy, but the toxicity was high and carried a risk of death. PMID- 17125481 TI - The pathophysiology of lower urinary tract symptoms after brachytherapy for prostate cancer. AB - OBJECTIVES: To determine the spectrum of pathophysiology underlying the lower urinary tract symptoms (LUTS) persisting for > or = 6 months after brachytherapy for localized prostate cancer. PATIENTS AND METHODS: A database of men from two practice settings was searched for men who developed LUTS persisting for > or = 6 months after completing brachytherapy for localized prostate cancer. Patients were evaluated with a structured history and physical examination, International Prostate Symptom Score (IPSS), 24-h voiding diary, noninvasive free-flow uroflowmetry, postvoid residual urine volume (PVR), cystoscopy and a video urodynamic study. Specific data collected included symptoms, elapsed time since brachytherapy, Gleason score, IPSS, total number of voids/24 h, maximum voided volume, cystoscopic findings, and urodynamics findings (PVR, maximum urinary flow rate, Schaefer obstruction grade, Watts factor, incidence of detrusor overactivity (DO) urethral obstruction and low bladder compliance). These data were compared with those from a previous study of men with LUTS who did not have prostate cancer. RESULTS: The study included 47 men (aged 54-88 years); the median (range) interval between brachytherapy and evaluation was 1.5 (0.5-13) years. Thirty-seven men complained of overactive bladder symptoms (79%), and 31 of incontinence (71%), 21 of obstructive symptoms (44%), and persistent dysuria in 12 (26%). Comparison of urodynamic findings in men with unselected causes of LUTS vs LUTS due to brachytherapy revealed the following comparisons: DO in 252 of 541 (47%) unselected vs 28 of 33 (85%) brachytherapy, (P < 0.001); and urethral obstruction in 374 of 541 (69%) unselected vs 24 of 33 (73%) brachytherapy (P = 0.85). CONCLUSION: The pathophysiology and severity of persistent LUTS in men after brachytherapy differs from that of men with LUTS in the general population. Men after brachytherapy have a much higher incidence of DO, prostatic and urethral strictures and prostatic urethral stones. PMID- 17125482 TI - An aqueous garlic extract alleviates water avoidance stress-induced degeneration of the urinary bladder. AB - OBJECTIVE: To investigate the role of aqueous garlic extract (AGE) on the water avoidance stress (WAS)-induced degeneration of the urinary bladder in a rat model. MATERIALS AND METHODS: Wistar albino rats were exposed to WAS for 2 h/day for 5 days (WAS group), after which, AGE (1 mL/kg) was injected intraperitoneally into the rats (WAS + AGE group). Urinary bladder samples were investigated with both light and scanning electron microscopy, and lipid peroxidation and glutathione levels were also measured in the samples. RESULTS: In the WAS group there was inflammatory cell infiltration, more mast cells and ulcerated areas in the mucosa. In the WAS + AGE group there was relatively normal urothelial alignment, moderate inflammatory cell infiltration and fewer mast cells in the mucosa. The increased lipid peroxidation and decreased glutathione levels in WAS rats were reversed by AGE treatment. CONCLUSIONS: These results show that AGE has a protective effect on WAS-induced degenerative changes in the urinary bladder. PMID- 17125483 TI - Early use of a phosphodiesterase inhibitor after brachytherapy restores and preserves erectile function. AB - OBJECTIVE: To investigate whether the early use of phosphodiesterase inhibitors (PDEIs) after brachytherapy (BT) is associated with better erectile function, as of men potent before BT 38-70% have erectile dysfunction afterward. PATIENTS AND METHODS: We evaluated a prospectively created database of 2500 patients who had had BT at our institution since 1992. We measured baseline age, cancer stage, Gleason grade, prostate specific antigen (PSA) level at diagnosis, implant type, use of neoadjuvant and adjuvant hormonal suppression therapy, use of external beam radiotherapy in conjunction with interstitial therapy, and follow-up PSA levels. Men were stratified by their use of PDEIs at <1 year (early group) or >1 year after implantation (late group). We excluded all men who did not have baseline Sexual Health Inventory for Men (SHIM) scores and at least one follow-up SHIM score; the latter were obtained at 6-month intervals after BT. Data were analysed using the Mann-Whitney U-test. RESULTS: In all, 210 men met the inclusion criteria; 85 began using PDEIs within a year of BT, and 125 started after a year. The mean time to PDEI use was 191 days in the early and 595 days in the late group. The median age was 62 years in the early and 63 years in the late group (P = 0.02). Baseline Gleason scores did not differ, nor did PSA levels between the groups. Of men in the early group, 48% received neoadjuvant and/or adjuvant hormonal suppression therapy, vs half of men in the late group. Baseline SHIM scores were not significantly different, nor were scores at the first two follow-up assessments, but the scores at 18-36 months after BT were significantly different. CONCLUSION: The early use of PDEIs after BT is associated with a significant improvement in and maintenance of erectile function compared with late use. Men undergoing BT should be encouraged to use PDEIs early after implantation, to preserve erectile function. PMID- 17125484 TI - The role of prolactin levels in the sexual activity of married men with erectile dysfunction. AB - OBJECTIVE: To evaluate the effects of risk factors for erectile dysfunction (ED) including anthropometry, hormones, metabolic profiles and lifestyle, on sexual activity in married men with ED. PATIENTS AND METHODS: The study included 261 men (mean age 53.7 years, range 23-80), who were evaluated for anthropometry, hormone levels, metabolic profiles and lifestyle factors. Erectile function was evaluated using the self-administered International Index of Erectile Function. Patients were classified into two groups based on the six-item erectile-function domain, as those with sexual activity and those without. RESULTS: Of all patients, 62.5% (163 of 261) had no sexual activity (erectile-function domain score <6). There was a significant difference in mean (sem) prolactin level between patients with and with no sexual activity, at 4.8 (0.4) vs 6.8 (0.7) (P = 0.013). Of all patients, 73.7% (42 of 57) with diabetes had no sexual activity, while 59.3% (121 of 204) without diabetes had (P = 0.048). In a multivariate model, a higher prolactin level was associated with a greater likelihood of sexual inactivity (odds ratio 1.094; 95% confidence interval, 1.010-1.185; P = 0.028) but diabetes lost its statistical significance. The area under the receiver-operating characteristic curve for prolactin was 60.5% (95% confidence interval, 52.9 68.1%; P = 0.009) for sexual inactivity. No other factors were significant in this regard. CONCLUSION: Our findings suggest that prolactin levels might play a role in sexual activity in men with ED. PMID- 17125485 TI - Does obesity influence the operative course or complications of robot-assisted laparoscopic prostatectomy. AB - OBJECTIVE: To determine if obesity is associated with prolonged surgery or more complications during and after surgery in patients undergoing robot-assisted laparoscopic prostatectomy (RALP). PATIENTS AND METHODS: Between December 2003 and September 2005 data were collected prospectively for all patients undergoing RALP. Obesity was defined as a body mass index of > or = 30 kg/m2. The duration of surgery was recorded, determining the duration of steps, including overall, robotic assistance, and vesico-urethral anastomosis. In all, 285 patients had RALP by one of two surgeons; 236 were classified as not obese and 49 as obese. RESULTS: The characteristics of the two groups were similar before and after RALP (mean age 60.7 vs 60.3 years, prostate-specific antigen level 7.9 vs 7.9 ng/mL, prostate weight 47.3 vs 45.3 g, length of stay 3.2 vs 3.5 days, and indwelling catheterization 8.3 vs 7.9 days). The mean duration of surgical steps was less for the non-obese than the obese group, with urethral dissection requiring 16 vs 19.3 min (P < 0.05), anastomotic time 30 vs 36 min (P < 0.01), and port closure 11 vs 14.5 min (P < 0.001), but the total duration of surgery, at 190 vs 205 min, was not significantly different (P = 0.07). In the non-obese group there were seven (3%) prolonged drain tube leaks, vs four (8%) in the obese group (P = 0.08). CONCLUSIONS: Obesity was associated with longer anastomotic times, but the overall duration was similar. Overall complication rates were not significantly different. The continuing follow-up will determine if obesity predisposes patients to greater long-term morbidity after RALP. PMID- 17125486 TI - The contemporary management of renal and ureteric calculi. PMID- 17125488 TI - Circumventing side-effects. PMID- 17125489 TI - Nurse-led flexible cystoscopy: Experience from one UK centre. PMID- 17125490 TI - Hypoandrogen-metabolic syndrome: A significant issue for men's health. PMID- 17125491 TI - Identification of the retrotrigonal layer as a key anatomical landmark during robotically assisted radical prostatectomy. PMID- 17125492 TI - Identification of the retrotrigonal layer as a key anatomical landmark during robotically assisted radical prostatectomy. PMID- 17125493 TI - Pre-clinical evidence for the use of phosphodiesterase-5 inhibitors for treating benign prostatic hyperplasia and lower urinary tract symptoms. PMID- 17125494 TI - Surgical atlas varicocele: Antegrade scrotal sclerotherapy. PMID- 17125495 TI - CNS progenitor cells and oligodendrocytes are targets of chemotherapeutic agents in vitro and in vivo. AB - BACKGROUND: Chemotherapy in cancer patients can be associated with serious short- and long-term adverse neurological effects, such as leukoencephalopathy and cognitive impairment, even when therapy is delivered systemically. The underlying cellular basis for these adverse effects is poorly understood. RESULTS: We found that three mainstream chemotherapeutic agents--carmustine (BCNU), cisplatin, and cytosine arabinoside (cytarabine), representing two DNA cross-linking agents and an antimetabolite, respectively--applied at clinically relevant exposure levels to cultured cells are more toxic for the progenitor cells of the CNS and for nondividing oligodendrocytes than they are for multiple cancer cell lines. Enhancement of cell death and suppression of cell division were seen in vitro and in vivo. When administered systemically in mice, these chemotherapeutic agents were associated with increased cell death and decreased cell division in the subventricular zone, in the dentate gyrus of the hippocampus and in the corpus callosum of the CNS. In some cases, cell division was reduced, and cell death increased, for weeks after drug administration ended. CONCLUSION: Identifying neural populations at risk during any cancer treatment is of great importance in developing means of reducing neurotoxicity and preserving quality of life in long term survivors. Thus, as well as providing possible explanations for the adverse neurological effects of systemic chemotherapy, the strong correlations between our in vitro and in vivo analyses indicate that the same approaches we used to identify the reported toxicities can also provide rapid in vitro screens for analyzing new therapies and discovering means of achieving selective protection or targeted killing. PMID- 17125496 TI - Efficacy of three artemisinin combination therapies for the treatment of uncomplicated Plasmodium falciparum malaria in the Republic of Congo. AB - BACKGROUND: Presented here are the results of a comparative trial on the efficacy of three artemisinin-based combinations conducted from May to October 2004, in Pool Province, Republic of Congo. METHODS: The main outcome was the proportion of cases of true treatment success at day 28. Recrudescences were distinguished from re-infections by PCR analysis. A total of 298 children of 6-59 months were randomized to receive either artesunate + SP (AS+SP), artesunate + amodiaquine (AS+AQ) or artemether + lumefantrine (AL), of which 15 (5%) were lost to follow up. RESULTS: After 28 days, there were 21/85 (25%) recurrent parasitaemias in the AS+SP group, 31/97 (32%) in the AS+AQ group and 13/100 (13%) in the AL group. The 28-day PCR-corrected cure rate was 90.1% [95% CI 80.7-95.9] for AS+SP, 98.5% [95% CI 92.0-100] for AS+AQ and 100% [95.8-100] for AL, thereby revealing a weaker response to AS+SP than to AL (p = 0.003) and to AS+AQ (p = 0.06). A potential bias was the fact that children treated with AL were slightly older and in better clinical condition, but logistic regression did not identify these as relevant factors. There was no significant difference between groups in fever and parasite clearance time, improvement of anaemia and gametocyte carriage at day 28. No serious adverse events were reported. CONCLUSION: Considering the higher efficacy of AL as compared to AS+SP and the relatively high proportion of cases with re infections in the AS+AQ group, we conclude that AL is clinically more effective than AS+SP and AS+AQ in this area of the Republic of Congo. Implementation of the recently chosen new national first-line AS+AQ should be monitored closely. PMID- 17125497 TI - A study of association between common variation in the growth hormone-chorionic somatomammotropin hormone gene cluster and adult fasting insulin in a UK Caucasian population. AB - BACKGROUND: Reduced growth during infancy is associated with adult insulin resistance. In a UK Caucasian cohort, the CSH1.01 microsatellite polymorphism in the growth hormone-chorionic somatomammotropin hormone gene cluster was recently associated with increases in adult fasting insulin of approximately 23 pmol/l for TT homozygote males compared to D1D1 or D2D2 homozygotes (P = 0.001 and 0.009; n = 206 and 92, respectively), but not for females. TT males additionally had a 547 g lower weight at 1 year (n = 270; P = 0.008) than D2D2 males. We sought to replicate these data in healthy UK Caucasian subjects. We genotyped 1396 subjects (fathers, mothers and children) from a consecutive birth study for the CSH1.01 marker and analysed genotypes for association with 1-year weight in boys and fasting insulin in fathers. RESULTS: We found no evidence for association of CSH1.01 genotype with adult male fasting insulin concentrations (TT/D1D1 P = 0.38; TT/D2D2 P = 0.18) or weight at 1 year in boys (TT/D1D1 P = 0.76; TT/D2D2 P = 0.85). For fasting insulin, our data can exclude the previously observed effect sizes as the 95 % confidence intervals for the differences observed in our study exclude increases in fasting insulin of 9.0 and 12.6 pmol/l for TT relative to D1D1 and D2D2 homozygotes, respectively. Whilst we have fewer data on boys' 1 year weight than the original study, our data can exclude a reduction in 1-year weight greater than 557 g for TT relative to D2D2 homozygotes. CONCLUSION: We have not found association of the CSH1.01 genotype with fasting insulin or weight at 1 year. We conclude that the original study is likely to have over-estimated the effect size for fasting insulin, or that the difference in results reflects the younger age of subjects in this study relative to those in the previous study. PMID- 17125498 TI - Clinicopathological analysis of histological variants of ameloblastoma in a suburban Nigerian population. AB - BACKGROUND: This study was carried out to establish the relative incidence and provide clinico-pathologic information on the various histological types of ameloblastoma seen at the Obafemi Awolowo University Teaching Hospital complex, Ile-Ife in order to provide a baseline data which will be of significance to the pathologist and clinician. METHODS: Clinico-pathologic data on a total of 77 histologically diagnosed cases of ameloblastoma archieved at the Obafemi Awolowo University Teaching Hospital Complex, Ile-Ife over a 15 year period were obtained and analysed descriptively. RESULTS: Follicular ameloblastoma was the most common histological type (50 cases, 64.9%), followed by plexiform ameloblastoma (10 cases, 13.0%). 4 (5.2%) cases of desmoplastic and 3 (3.9%) cases of acanthomatous ameloblastoma were seen while the basal cell variant accounted for 2 (2.6%) cases. Only 1 case of the unicystic type was seen. Some of the 77 cases presented as a mixture of two or more histological types. Ameloblastoma occurred over an age range of 11 to 70 years with a peak age incidence in the 3rd decade. CONCLUSION: This study provides a baseline data on variants of ameloblastoma as obtained in a suburban Nigerian population. Since variants of ameloblastoma differ in biologic behaviour, the data collected in this study provides clinicopathologic information which is of significance to the pathologist and clinician. PMID- 17125499 TI - The role of crude human saliva and purified salivary MUC5B and MUC7 mucins in the inhibition of Human Immunodeficiency Virus type 1 in an inhibition assay. AB - BACKGROUND: Despite the continuous shedding of HIV infected blood into the oral cavity and the detectable presence of the AIDS virus at a high frequency, human saliva is reported to inhibit oral transmission of HIV through kissing, dental treatment, biting, and aerosolization. The purpose of this study was to purify salivary MUC5B and MUC7 mucins from crude saliva and determine their anti-HIV-1 activities. METHODS: Following Sepharose CL-4B column chromatography and caesium chloride isopycnic density-gradient ultra-centrifugation, the purity and identity of the mucins was determined by SDS-PAGE and Western blotting analysis respectively. Subsequently an HIV-1 inhibition assay was carried out to determine the anti-HIV-1 activity of the crude saliva and purified salivary mucins by incubating them with subtype D HIV-1 prior to infection of the CD4+ CEM SS cells. RESULTS: Western blotting analysis confirmed that the mucin in the void volume is MUC5B and the mucin in the included volume is MUC7. The HIV inhibition assay revealed that both the crude saliva and salivary MUC5B and MUC7 mucins inhibited HIV-1 activity by 100%. CONCLUSION: Although the mechanism of action is not clear the carbohydrate moieties of the salivary mucins may trap or aggregate the virus and prevent host cell entry. PMID- 17125500 TI - Totally laparoscopic versus conventional ileoanal pouch procedure--design of a single-centre, expertise based randomised controlled trial to compare the laparoscopic and conventional surgical approach in patients undergoing primary elective restorative proctocolectomy--LapConPouch-Trial. AB - BACKGROUND: Restorative proctocolectomy is increasingly being performed minimal invasively but a totally laparoscopic technique has not yet been compared to the standard open technique in a randomized study. METHODS/DESIGN: This is a two armed, single centre, expertise based, preoperatively randomized, patient blinded study. It is designed as a two-group parallel superiority study. Power calculation revealed 80 patients per group in order to recruit the 65 patients to be analysed for the primary endpoint. The primary objective is to investigate intra-operative blood loss and the need for blood transfusions. We hypothesise that intra-operative blood loss and the need for peri-operative blood transfusions are significantly higher in the conventional group. Additionally a set of surgical and non-surgical parameters related to the operation will be analysed as secondary objectives. These will include operative time, complications, postoperative pain, lung function, postoperative length of hospital stay, a cosmetic score and pre-and postoperative quality of life. DISCUSSION: The trial will answer the question whether there is indeed an advantage in the laparoscopic group in regard to blood loss and the need for blood transfusions. Moreover, it will generate data on the safety and potential advantages and disadvantages of the minimally invasive approach. PMID- 17125501 TI - Survival of immature Anopheles arabiensis (Diptera: Culicidae) in aquatic habitats in Mwea rice irrigation scheme, central Kenya. AB - BACKGROUND: The survivorship and distribution of Anopheles arabiensis larvae and pupae was examined in a rice agro-ecosystem in Mwea Irrigation Scheme, central Kenya, from August 2005 to April 2006, prior to implementation of larval control programme. METHODS: Horizontal life tables were constructed for immatures in semi field condition. The time spent in the various immature stages was determined and survival established. Vertical life tables were obtained from five paddies sampled by standard dipping technique. RESULTS: Pre-adult developmental time for An. arabiensis in the trays in the experimental set up in the screen house was 11.85 days from eclosion to emergence. The mean duration of each instar stage was estimated to be 1.40 days for first instars, 2.90 days for second instars, 1.85 days for third instars, 3.80 days for fourth instars and 1.90 days for pupae. A total of 590 individuals emerged into adults, giving an overall survivorship from L1 to adult emergence of 69.4%. A total of 4,956 An. arabiensis immatures were collected in 1,400 dips throughout the sampling period. Of these, 55.9% were collected during the tillering stage, 42.5% during the transplanting period and 1.6% during the land preparation stage. There was a significant difference in the An. arabiensis larval densities among the five stages. Also there was significant variation in immature stage composition for each day's collection in each paddy. These results indicate that the survival of the immatures was higher in some paddies than others. The mortality rate during the transplanting was 99.9% and at tillering was 96.6%, while the overall mortality was 98.3%. CONCLUSION: The survival of An. arabiensis immatures was better during the tillering stage of rice growth. Further the survival of immatures in rice fields is influenced by the rice agronomic activities including addition of nitrogenous fertilizers and pesticides. For effective integrated vector management, the application of larvicides should target An. arabiensis larvae at the tillering stage (early vegetative stage of rice) when their survival in the aquatic habitats is high to significantly reduce them and the larvicides should be long-lasting to have a significant impact on the malaria vector productivity on the habitats. PMID- 17125502 TI - Acceptability and outcomes of the Percutaneous Endoscopic Gastrostomy (PEG) tube placement--patients' and care givers' perspectives. AB - BACKGROUND: Percutaneous endoscopic gastrostomy tube has now become a preferred option for the long-term nutritional support device for patients with dysphagia. There is a considerable debate about the health issues related to the quality of life of these patients. Our aim of the study was to assess the outcome and perspectives of patients/care givers, about the acceptability of percutaneous endoscopic gastrostomy tube placement. METHODS: This descriptive analytic study conducted in patients, who have undergone percutaneous endoscopic gastrostomy tube placement during January 1998 till December 2004. Medical records of these patients were evaluated for their demographic characteristics, underlying diagnosis, indications and complications. Telephonic interviews were conducted till March 2005, on a pre-tested questionnaire to address psychological, social and physical performance status, of the health related quality of life issues. RESULTS: A total of 191 patients' medical records were reviewed, 120 (63%) were males, and mean age was 63 years. Early complication was infection at PEG tube site in 6 (3%) patients. In follow up over 365 +/- 149 days, late complications (occurring 72 hours later) were infection at PEG tube site in 29 (15 %) patient and dislodgment/blockage of the tube in 26 (13.6%). Interviews were possible with 126 patients/caretakers. Karnofsky Performance Score of 0, 1, 2, 3 and 4 was found in 13(10%), 18(14%), 21(17%), 29(23%) and 45(36%) with p-value < 0.001. Regarding the social and psychological aspects; 76(60%) would like to have the PEG tube again if required, 105(83 %) felt ease in feeding, and 76(60%) felt that PEG-tube helped in prolonging the survival. Regarding negative opinions; 49(39 %) felt that the feeding was too frequent, 45(36 %) felt apprehensive about dependency for feeding and 62(49%) were concerned about an increase in the cost of care. CONCLUSION: PEG-tube placement was found to be relatively free from serious immediate and long- term complications. Majority of caregivers and patient felt that PEG-tube helped in feeding and prolonging the survival. Studies are needed to assess the real benefit in terms of actual nutritional gain and quality of life in such patients. PMID- 17125503 TI - Children undergoing cardiac surgery for complex cardiac defects show imbalance between pro- and anti-thrombotic activity. AB - INTRODUCTION: Cardiac surgery with cardiopulmonary bypass (CPB) is associated with the activation of inflammatory mediators that possess prothrombotic activity and could cause postoperative haemostatic disorders. This study was conducted to investigate the effect of cardiac surgery on prothrombotic activity in children undergoing cardiac surgery for complex cardiac defects. METHODS: Eighteen children (ages 3 to 163 months) undergoing univentricular palliation with total cavopulmonary connection (TCPC) (n = 10) or a biventricular repair (n = 8) for complex cardiac defects were studied. Prothrombotic activity was evaluated by measuring plasma levels of prothrombin fragment 1+2 (F1+2), thromboxane B2 (TxB2), and monocyte chemoattractant protein-1 (MCP-1). Anti-thrombotic activity was evaluated by measuring levels of tissue factor pathway inhibitor (TFPI) before, during, and after cardiac surgery. RESULTS: In all patients, cardiac surgery was associated with a significant but transient increase of F1+2, TxB2, TFPI, and MCP-1. Maximal values of F1+2, TxB2, and MCP-1 were found at the end of CPB. In contrast, maximal levels of TFPI were observed at the beginning of CPB. Concentrations of F1+2 at the end of CPB correlated negatively with the minimal oesophageal temperature during CPB. Markers of prothrombotic activity returned to preoperative values from the first postoperative day on. Early postoperative TFPI levels were significantly lower and TxB2 levels significantly higher in patients with TCPC than in those with biventricular repair. Thromboembolic events were not observed. CONCLUSION: Our data suggest that children with complex cardiac defects undergoing cardiac surgery show profound but transient imbalance between pro- and anti-thrombotic activity, which could lead to thromboembolic complications. These alterations are more important after TCPC than after biventricular repair but seem to be determined mainly by low antithrombin III. PMID- 17125504 TI - Musculoskeletal disorders in shipyard industry: prevalence, health care use, and absenteeism. AB - BACKGROUND: It is unclear whether the well-known risk factors for the occurrence of musculoskeletal disorders (MSD) also play an important role in the determining consequences of MSD in terms of sickness absence and health care use. METHODS: A cross-sectional study was conducted among 853 shipyard employees. Data were collected by questionnaire on physical and psychosocial workload, need for recovery, perceived general health, occurrence of musculoskeletal complaints, and health care use during the past year. Retrospective data on absenteeism were also available from the company register. RESULTS: In total, 37%, 22%, and 15% of employees reported complaints of low back, shoulder/neck, and hand/wrist during the past 12 months, respectively. Among all employees with at least one MSD, 27% visited a physician at least once and 20% took at least one period of sick leave. Various individual and work-related factors were associated with the occurrence of MSD. Health care use and absenteeism were strongest influenced by chronicity of musculoskeletal complaints and comorbidity with other musculoskeletal complaints and, to a lesser extent, by work-related factors. CONCLUSION: In programmes aimed at preventing the unfavourable consequences of MSD in terms of sickness absence and health care use it is important to identify the (individual) factors that determine the development of chronicity of complaints. These factors may differ from the well-know risk factors for the occurrence of MSD that are targeted in primary prevention. PMID- 17125505 TI - EDAR mutation in autosomal dominant hypohidrotic ectodermal dysplasia in two Swedish families. AB - BACKGROUND: Hypohidrotic ectodermal dysplasia (HED) is a genetic disorder characterized by defective development of teeth, hair, nails and eccrine sweat glands. Both autosomal dominant and autosomal recessive forms of HED have previously been linked to mutations in the ectodysplasin 1 anhidrotic receptor (EDAR) protein that plays an important role during embryogenesis. METHODS: The coding DNA sequence of the EDAR gene was analyzed in two large Swedish three generational families with autosomal dominant HED. RESULTS: A non-sense C to T mutation in exon 12 was identified in both families. This disease-specific mutation changes an arginine amino acid in position 358 of the EDAR protein into a stop codon (p.Arg358X), thereby truncating the protein. In addition to the causative mutation two polymorphisms, not associated with the HED disorder, were also found in the EDAR gene. CONCLUSION: The finding of the p.Arg358X mutation in the Swedish families is the first corroboration of a previously described observation in an American family. Thus, our study strengthens the role of this particular mutation in the aetiology of autosomal dominant HED and confirms the importance of EDAR for the development of HED. PMID- 17125506 TI - Cough induced rib fracture, rupture of the diaphragm and abdominal herniation. AB - Cough can be associated with many complications. In this article, we present a 59 year old male patient with a very rare combination of a cough related stress fracture of the ninth rib, a traumatic rupture of the diaphragm, and an abdominal wall herniation. The hernia was repaired through surgical treatment without bowel resection, the diaphragm and the internal and oblique abdominal muscle were adapted, and the abdomen was reinforced with a prolene net. Although each individual injury is well documented in the literature, the combination of rib fracture, abdominal herniation and diaphragm rupture has not been reported. PMID- 17125507 TI - Determinants of fruit and vegetable intake among 11-year-old schoolchildren in a country of traditionally low fruit and vegetable consumption. AB - BACKGROUND: Fruit and vegetable consumption is traditionally low in Iceland. The results of the Pro Children cross-Europe survey showed that the consumption was lowest among children in Iceland. The aim of this study was to identify determinants of fruit and vegetable intake among 11-year-old schoolchildren in Iceland. METHODS: A cross-sectional survey was performed in Iceland in the autumn of 2003 as a part of the Pro Children cross-Europe survey. The survey was designed to provide information on actual consumption levels of vegetables and fruits by 11-year-old school children and to assess potential determinants of consumption patterns. A total of 1235 Icelandic children (89%) from 32 randomly chosen schools participated. Hierarchical regression analyses were performed to determine the explained variance of the children's fruit and vegetable intake. In these analyses socio-demographic background variables were entered as a first block, perceived physical-environmental variables as a second block, perceived socio-environmental variables as a third block and personal variables as a fourth block. RESULTS: 64% of the children ate fruit less than once a day, and 61% ate vegetables less than once a day. Respectively, 31% and 39% of the variance in children's fruit and vegetable intake was explained by the determinants studied. About 7% and 13% of the variance in fruit and vegetable intake was explained by the perceived physical-environmental determinants, mainly by availability at home. About 18% and 16% of the variance in fruit and vegetable intake was explained by the personal determinants. For both fruit and vegetable intake, the significant personal determinants were preferences, liking, knowledge of recommendations and self-efficacy. CONCLUSION: Interventions to increase fruit and vegetable intake among children should aim at both environmental factors such as greater availability of fruit and vegetables, and personal factors as self efficacy and knowledge levels concerning nutrition. PMID- 17125508 TI - Histological analysis of the effects of a static magnetic field on bone healing process in rat femurs. AB - BACKGROUND: The aim of this study was to investigate, in vivo, the quality of bone healing under the effect of a static magnetic field, arranged inside the body. METHODS: A metallic device was developed, consisting of two stainless steel washers attached to the bone structure with titanium screws. Twenty-one Wistar rats (Rattus novergicus albinus) were used in this randomized experimental study. Each experimental group had five rats, and two animals were included as control for each of the groups. A pair of metal device was attached to the left femur of each animal, lightly touching a surgically created bone cavity. In the experimental groups, washers were placed in that way that they allowed mutual attraction forces. In the control group, surgery was performed but washers, screws or instruments were not magnetized. The animals were sacrificed 15, 45 and 60 days later, and the samples were submitted to histological analysis. RESULTS: On days 15 and 45 after the surgical procedure, bone healing was more effective in the experimental group as compared to control animals. Sixty days after the surgical procedure, marked bone neoformation was observed in the test group, suggesting the existence of continued magnetic stimulation during the experiment. CONCLUSION: The magnetic stainless steel device, buried in the bone, in vivo, resulted in increased efficiency of the experimental bone healing process. PMID- 17125509 TI - Hypnotic use in a population-based sample of over thirty-five thousand interviewed Canadians. AB - BACKGROUND: As with most medications, benzodiazepine and similar sedative hypnotics (BDZ/SSH) can produce both beneficial and adverse effects. Pharmacoepidemiological studies have been limited in their capacity to evaluate the relationship between these medications and psychiatric diagnoses in non clinical populations. The objective of this study was to provide a description of the pattern of use of BDZ/SSH medications in relation to both demographic and diagnostic data in a community population. METHODS: The source of data for this study was the Canadian Community Health Survey (CCHS 1.2), also known as the Canadian National Study of Mental Health and Well-being. This study was based on a nationally representative sample that included over 35 thousand subjects with a response rate of 77%. The survey interview included the latest version of the Composite International Diagnostic Interview (CIDI), which was developed for the World Health Organization's WHO Mental Health 2000 project. Current medication use was also recorded. RESULTS: As expected, BDZ/SSH use was more common in women than in men (4.2%, 95% CI 3.9 to 4.6 vs. 2.5%, 95% CI 2.2 to 2.8) and its frequency increased with age, 8.5% (95% CI 7.7 to 9.4) of those over the age of 65 compared to 2.4% (95% CI 2.2 to 2.7) of those aged 18 to 64 years. These medications were more frequently used in subjects with low levels of education (4.8%, 95% CI% 4.3 to 5.2) vs. high levels of education (2.4%, 95% CI 2.1 to 2.6) and low personal incomes (5.7%, 95% CI 5.2 to 6.3) vs. high personal incomes (2.3%, 95% CI 2.0 to 2.6). BDZ/SSH use was strongly associated with the presence of mood or anxiety disorders, but not with substance use disorders. Demographic differences persisted after statistical adjustment for diagnosis. CONCLUSION: The observation that benzodiazepine use is more frequent in women, increases with age and is higher in low income and education groups supports previous findings. These results help to confirm that these differences are not accounted for by psychiatric diagnoses. PMID- 17125510 TI - Comparison of human uterine cervical electrical impedance measurements derived using two tetrapolar probes of different sizes. AB - BACKGROUND: We sought to compare uterine cervical electrical impedance spectroscopy measurements employing two probes of different sizes, and to employ a finite element model to predict and compare the fraction of electrical current derived from subepithelial stromal tissue. METHODS: Cervical impedance was measured in 12 subjects during early pregnancy using 2 different sizes of the probes on each subject. RESULTS: Mean cervical resistivity was significantly higher (5.4 vs. 2.8 Omegam; p < 0.001) with the smaller probe in the frequency rage of 4-819 kHz. There was no difference in the short-term intra-observer variability between the two probes. The cervical impedance measurements derived in vivo followed the pattern predicted by the finite element model. CONCLUSION: Inter-electrode distance on the probes for measuring cervical impedance influences the tissue resistivity values obtained. Determining the appropriate probe size is necessary when conducting clinical studies of resistivity of the cervix and other human tissues. PMID- 17125511 TI - Effective population size of Anopheles funestus chromosomal forms in Burkina Faso. AB - BACKGROUND: As Anopheles funestus is one of the principal Afro-tropical malaria vectors, a more complete understanding of its population structure is desirable. In West and Central Africa, An. funestus population structure is complicated by the coexistence of two assortatively mating chromosomal forms. Effective population size (Ne) is a key parameter in understanding patterns and levels of intraspecific variation, as it reflects the role of genetic drift. Here, Ne was estimated from both chromosomal forms, Kiribina and Folonzo, in Burkina Faso. METHODS: Short-term Ne was estimated by evaluating variation at 16 microsatellite loci across temporal samples collected annually from 2000-2002. Estimates were based on standardized variance in allele frequencies or a maximum likelihood method. Long-term Ne was estimated from genetic diversity estimates using mtDNA sequences and microsatellites. RESULTS: For both forms, short-term and long-term Ne estimates were on the order of 10(3) and 10(5), respectively. Long-term Ne estimates were larger when based on loci from chromosome 3R (both inside and outside of inversions) than loci outside of this arm. CONCLUSION: Ne values indicate that An. funestus is not subject to seasonal bottlenecks. Though not statistically different because of large and overlapping confidence intervals, short-term Ne estimates were consistently smaller for Kiribina than Folonzo, possibly due to exploitation of different breeding sites: permanent for Folonzo and intermittent for Kiribina. The higher long-term Ne estimates on 3R, the arm carrying the two inversions mainly responsible for defining the chromosomal forms, give natural selection broader scope and merit further study. PMID- 17125512 TI - Assessing responsiveness of generic and specific health related quality of life measures in heart failure. AB - BACKGROUND: Responsiveness, or sensitivity to clinical change, is an important consideration in selection of a health-related quality of life (HRQL) measure for trials or clinical applications. Many approaches can be used to assess responsiveness, which may affect the interpretation of study results. We compared the relative responsiveness of generic and heart failure specific HRQL instruments, as measured both by common psychometric indices and by external clinical criteria. METHODS: We analyzed data collected at baseline and 6-weeks in 298 subjects with heart failure on the following HRQL measures: EQ-5D (US, UK, and VAS Scoring), Kansas City Cardiomyopathy Questionnaire (KCCQ) (Clinical and Overall Summary Score), and RAND12 (Physical and Mental Component Summaries). Three external indicators of clinical change were used to classify subjects as improved, deteriorated, or unchanged: 6-minute walk test, New York Heart Association (NYHA) class, and physician global rating of change. Four responsiveness statistics (T-test, effect size, Guyatt's responsiveness statistic, and standardized response mean) were used to evaluate the responsiveness of the select measures. The median rank of each HRQL measure across responsiveness indices and clinical criteria was then determined. RESULTS: Average age of subjects was 60 years, 75 percent were male, and had moderate to severe heart failure symptoms. Overall, the KCCQ Summary Scores had the highest relative ranking, irrespective of the responsiveness index or external criterion used. Importantly, we observed that the relative ranking of responsiveness of the generic measures (i.e. EQ-5D, RAND12) was influenced by both the responsive indices and external criterion used. CONCLUSION: The disease specific KCCQ was the most responsive HRQL measure assessing change over a 6-week period, although generic measures provide information for which the KCCQ is not suitable. The responsiveness of generic HRQL measures may be affected by the index used, as well as the external criterion to classify patients who have clinically change or remained stable. PMID- 17125513 TI - Nuclear Factor 90, a cellular dsRNA binding protein inhibits the HIV Rev-export function. AB - BACKGROUND: The HIV Rev protein is known to facilitate export of incompletely spliced and unspliced viral transcripts to the cytoplasm, a necessary step in virus life cycle. The Rev-mediated nucleo-cytoplasmic transport of nascent viral transcripts, dependents on interaction of Rev with the RRE RNA structural element present in the target RNAs. The C-terminal variant of dsRNA-binding nuclear protein 90 (NF90ctv) has been shown to markedly attenuate viral replication in stably transduced HIV-1 target cell line. Here we examined a mechanism of interference of viral life cycle involving Rev-NF90ctv interaction. RESULTS: Since Rev:RRE complex formations depend on protein:RNA and protein:protein interactions, we investigated whether the expression of NF90ctv might interfere with Rev-mediated export of RRE-containing transcripts. When HeLa cells expressed both NF90ctv and Rev protein, we observed that NF90ctv inhibited the Rev-mediated RNA transport. In particular, three regions of NF90ctv protein are involved in blocking Rev function. Moreover, interaction of NF90ctv with the RRE RNA resulted in the expression of a reporter protein coding sequences linked to the RRE structure. Moreover, Rev influenced the subcellular localization of NF90ctv, and this process is leptomycin B sensitive. CONCLUSION: The dsRNA binding protein, NF90ctv competes with HIV Rev function at two levels, by competitive protein:protein interaction involving Rev binding to specific domains of NF90ctv, as well as by its binding to the RRE-RNA structure. Our results are consistent with a model of Rev-mediated HIV-1 RNA export that envisions Rev-multimerization, a process interrupted by NF90ctv. PMID- 17125514 TI - A hierarchical Naive Bayes Model for handling sample heterogeneity in classification problems: an application to tissue microarrays. AB - BACKGROUND: Uncertainty often affects molecular biology experiments and data for different reasons. Heterogeneity of gene or protein expression within the same tumor tissue is an example of biological uncertainty which should be taken into account when molecular markers are used in decision making. Tissue Microarray (TMA) experiments allow for large scale profiling of tissue biopsies, investigating protein patterns characterizing specific disease states. TMA studies deal with multiple sampling of the same patient, and therefore with multiple measurements of same protein target, to account for possible biological heterogeneity. The aim of this paper is to provide and validate a classification model taking into consideration the uncertainty associated with measuring replicate samples. RESULTS: We propose an extension of the well-known Naive Bayes classifier, which accounts for biological heterogeneity in a probabilistic framework, relying on Bayesian hierarchical models. The model, which can be efficiently learned from the training dataset, exploits a closed-form of classification equation, thus providing no additional computational cost with respect to the standard Naive Bayes classifier. We validated the approach on several simulated datasets comparing its performances with the Naive Bayes classifier. Moreover, we demonstrated that explicitly dealing with heterogeneity can improve classification accuracy on a TMA prostate cancer dataset. CONCLUSION: The proposed Hierarchical Naive Bayes classifier can be conveniently applied in problems where within sample heterogeneity must be taken into account, such as TMA experiments and biological contexts where several measurements (replicates) are available for the same biological sample. The performance of the new approach is better than the standard Naive Bayes model, in particular when the within sample heterogeneity is different in the different classes. PMID- 17125515 TI - Resistance to penicillin of Staphylococcus aureus isolates from cows with high somatic cell counts in organic and conventional dairy herds in Denmark. AB - BACKGROUND: Quarter milk samples from cows with high risk of intramammary infection were examined to determine the prevalence of Staphylococcus aureus (SA) and penicillin resistant SA (SAr) in conventional and organic dairy herds and herds converting to organic farming in a combined longitudinal and cross sectional study. METHODS: 20 conventional herds, 18 organic herds that converted before 1995, and 19 herds converting to organic farming in 1999 or 2000 were included in the study. Herds converting to organic farming were sampled three times one year apart; the other herds were sampled once. Risk of infection was estimated based on somatic cell count, milk production, breed, age and lactation stage. RESULTS: The high-risk cows represented about 49 % of the cows in the herds. The overall prevalence of SA and SAr among these cows was 29% (95% confidence interval: 24%-34%) and 4% (95% confidence interval: 2%-5%) respectively. The prevalence of penicillin resistance among SA infected cows was 12% (95% confidence interval: 6%-19%) when calculated from the first herd visits. No statistically significant differences were observed in the prevalence of SAr or the proportion of isolates resistant to penicillin between herd groups. CONCLUSION: The proportion of isolates resistant to penicillin was low compared to studies in other countries except Norway and Sweden. Based on the low prevalence of penicillin resistance of SA, penicillin should still be the first choice of antimicrobial agent for treatment of bovine intramammary infection in Denmark. PMID- 17125516 TI - Expression of EpCAM in uveal melanoma. AB - BACKGROUND: Uveal melanoma (UM) is the most common primary intraocular malignant tumor in adults, and nearly 40% of UM will develop metastasis that will ultimately lead to death. The Epithelial Cell Adhesion Molecule (EpCAM) is a type I transmembrane glycoprotein expressed by carcinomas of head and neck, ovary, colon, breast, kidney and lung. Recently, antibodies against EpCAM such as Edrecolomab and Catumaxomab were developed, and clinical trials with these antibodies have been used in several types of neoplasia. We studied the expression of EpCAM in UM. METHODS: 25 enucleated formalin-fixed, paraffin embedded UM specimens were immunostained for EpCAM. Histopathological analysis of the specimens with regards to prognostic factors such as cell type, largest (linear) tumor dimension, number of mitotic figures, scleral invasion and tumor infiltrating lymphocytes were done. RESULTS: None of them was positive for this EpCAM. CONCLUSION: In our report, UM did not express EpCAM. Therefore, it is not a helpful immunohistochemical marker to predict the behavior of UM. Further studies are needed to verify if EpCAM could also be related with prognosis and treatment of UM. PMID- 17125517 TI - A biometric and ecologic comparison between Artemia from Mexico and Chile. AB - BACKGROUND: A preliminary biometric and ecologic database for the brine shrimp Artemia from Mexico and Chile is presented. The area abounds in small and seasonal ponds and large inland lakes, the latter mainly located in Mexico, although relatively large and isolated lakes are found in complex hydrological settings in pre-high plateau areas of Chile. This paper summarizes research efforts aimed at the localization, characterization, and evaluation of the aquaculture potential of Artemia populations in Mexico and Chile, which exhibit great habitat diversity (ponds, salterns, coastal lagoons, sea arms, coastal and inland lakes), contrasting weather conditions and different levels of isolation and human intervention. RESULTS: This study covered locations between 29 degrees north latitude (Baja California, Mexico) to 50 degrees south latitude (Puerto Natales, Chile). Biological characteristics considered are species name, reproductive mode, cyst diameter, chorion thickness, and nauplius length, whereas ecological data include pond size, pH, salinity, temperature, and water ionic composition. Artemia franciscana is the only species found in Mexico, it exists together with A. persimilis in Chile, though separated geographically. Ecological differences in habitat exist between both regions but also within countries, a pattern particularly clear with regard to water composition. Surprisingly, a Mexican (Cuatro Cienegas, A. franciscana) and a Chilean location (Torres del Paine, A. persimilis) share habitat characteristics, at least for the period when data were collected. The discriminant analysis for cyst diameter and nauplius length shows that Artemia from only one location match in cyst diameter with those from San Francisco Bay (SFB) (Point Lobos), and one (Marquez) is far apart from SFB and all the others. The Chilean locations (Pampilla, Cejar, Cahuil, Llamara, Yape) share cyst diameter, but tend to differ from SFB. The remaining Mexican locations (Juchitan, Ohuira, Yavaros) are well separated from all the others. With regard to nauplii length, populations tend to distribute in a relatively random manner, being Marquez the location differing the most in cyst diameter from SFB. CONCLUSION: This database will contribute to the knowledge of radiation centers and serves as a baseline for further biogeographic studies, population characterization, management, and monitoring of Artemia biodiversity. Likewise, the impact of colonization and translocations for aquaculture purposes can be better assessed with a baseline for reference. Mexico and Chile exemplify the need to increase and further integrate regional information to tackle fundamental problems underlying practical utilization of Artemia. PMID- 17125518 TI - Genetic polymorphisms of MMP1, MMP3 and MMP7 gene promoter and risk of colorectal adenoma. AB - BACKGROUND: Matrix metalloproteinases (MMP) have been shown to play a role in colorectal cancer (CRC). More recently, MMP1, MMP3 and MMP7 functional gene promoter polymorphisms have been found to be associated with CRC occurrence and prognosis. To document the role of MMP polymorphisms in the early step of colorectal carcinogenesis, we investigated their association with colorectal adenoma risk in a case-control study comprising 295 patients with large adenomas (LA), 302 patients with small adenomas (SA) and 568 polyp-free (PF) controls. METHODS: Patients were genotyped using automated fragment analysis for MMP1 -1607 ins/del G and MMP3 -1612 ins/delA (MMP3.1) polymorphisms and allelic discrimination assay for MMP3 -709 A/G (MMP3.2) and MMP7 -181 A/G polymorphisms. Association between MMP genotypes and colorectal adenomas was first tested for each polymorphism separately and then for combined genotypes using the combination test. Adjustment on relevant variables and estimation of odds ratios were performed using unconditional logistic regression. RESULTS: No association was observed between the polymorphisms and LA when compared to PF or SA. When comparing SA to PF controls, analysis revealed a significant association between MMP3 -1612 ins/delA polymorphism and SA with an increased risk associated with the 6A/6A genotype (OR = 1.67, 95% CI: 1.20-2.34). Using the combination test, the best association was found for MMP3.1-MMP1 (p = 0.001) with an OR of 1.88 (95% CI: 1.08-3.28) for the combined genotype 2G/2G-6A/6A estimated by logistic regression. CONCLUSION: These data show a relation between MMP1 -1607 ins/del G and MMP3 -1612 ins/delA combined polymorphisms and risk of SA, suggesting their potential role in the early steps of colorectal carcinogenesis. PMID- 17125519 TI - Scanning electron microscopy of the neuropathology of murine cerebral malaria. AB - BACKGROUND: The mechanisms leading to death and functional impairments due to cerebral malaria (CM) are yet not fully understood. Most of the knowledge about the pathomechanisms of CM originates from studies in animal models. Though extensive histopathological studies of the murine brain during CM are existing, alterations have not been visualized by scanning electron microscopy (SEM) so far. The present study investigates the neuropathological features of murine CM by applying SEM. METHODS: C57BL/6J mice were infected with Plasmodium berghei ANKA blood stages. When typical symptoms of CM developed perfused brains were processed for SEM or light microscopy, respectively. RESULTS: Ultrastructural hallmarks were disruption of vessel walls, parenchymal haemorrhage, leukocyte sequestration to the endothelium, and diapedesis of macrophages and lymphocytes into the Virchow-Robin space. Villous appearance of observed lymphocytes were indicative of activated state. Cerebral oedema was evidenced by enlargement of perivascular spaces. CONCLUSION: The results of the present study corroborate the current understanding of CM pathophysiology, further support the prominent role of the local immune system in the neuropathology of CM and might expose new perspectives for further interventional studies. PMID- 17125520 TI - Socioeconomic status and hospital utilization among younger adult pneumonia admissions at a Canadian hospital. AB - BACKGROUND: Although the general association between socioeconomic status (SES) and hospitalization has been well established, few studies have considered the relationship between SES and hospital length of stay (LOS), and/or hospital re admission. The primary objective of this study therefore, was to examine the relationship of SES to LOS and early re-admission among adult patients hospitalized with community-acquired pneumonia in a setting with universal health insurance. METHODS: Four hundred and thirty-four (434) individuals were included in this retrospective, longitudinal cohort analysis of adult patients less than 65 years old admitted to a large teaching hospital in Vancouver, British Columbia. Hospital chart review data were linked to population-based health plan administrative data. Chart review was used to gather data on demographics, illness severity, co-morbidity, functional status and other measures of case mix. Two different types of administrative data were used to determine hospital LOS and the occurrence of all-cause re-admission to any hospital within 30 days of discharge. SES was measured by individual-level financial hardship (receipt of income assistance or provincial disability pension) and neighbourhood-level income quintiles. RESULTS: Those with individual-level financial hardship had an estimated 15% (95% CI -0.4%, +32%, p = 0.057) longer adjusted LOS and greater risk of early re-admission (adjusted OR 2.65, 95% CI 1.38, 5.09). Neighbourhood level income quintiles, showed no association with LOS or early re-admission. CONCLUSION: Among hospitalized pneumonia patients less than 65 years, financial hardship derived from individual-level data, was associated with an over two-fold greater risk of early re-admission and a marginally significant longer hospital LOS. However, the same association was not apparent when an ecological measure of SES derived from neighbourhood income quintiles was examined. The ecological SES variable, while useful in many circumstances, may lack the sensitivity to detect the full range of SES effects in clinical studies. PMID- 17125521 TI - Breastfeeding support for adolescent mothers: similarities and differences in the approach of midwives and qualified breastfeeding supporters. AB - BACKGROUND: The protection, promotion and support of breastfeeding are now major public health priorities. It is well established that skilled support, voluntary or professional, proactively offered to women who want to breastfeed, can increase the initiation and/or duration of breastfeeding. Low levels of breastfeeding uptake and continuation amongst adolescent mothers in industrialised countries suggest that this is a group that is in particular need of breastfeeding support. Using qualitative methods, the present study aimed to investigate the similarities and differences in the approaches of midwives and qualified breastfeeding supporters (the Breastfeeding Network (BfN)) in supporting breastfeeding adolescent mothers. METHODS: The study was conducted in the North West of England between September 2001 and October 2002. The supportive approaches of 12 midwives and 12 BfN supporters were evaluated using vignettes, short descriptions of an event designed to obtain specific information from participants about their knowledge, perceptions and attitudes to a particular situation. Responses to vignettes were analysed using thematic networks analysis, involving the extraction of basic themes by analysing each script line by line. The basic themes were then grouped to form organising themes and finally central global themes. Discussion and consensus was reached related to the systematic development of the three levels of theme. RESULTS: Five components of support were identified: emotional, esteem, instrumental, informational and network support. Whilst the supportive approaches of both groups incorporated elements of each of the five components of support, BfN supporters placed greater emphasis upon providing emotional and esteem support and highlighted the need to elicit the mothers' existing knowledge, checking understanding through use of open questions and utilising more tentative language. Midwives were more directive and gave more examples of closed questions. These differences could reflect the considerable emphasis upon person-centred approaches within the BfN curriculum and, in the case of midwives, the bureaucratic and institutional constraints upon them making it difficult, if not impossible, to take time and touch base with women. CONCLUSION: Follow up ethnographic work is required to assess the differences in the supportive approaches of BfN supporters and midwives in the practice areas. Such research, which specifically focuses upon how the different approaches are received and experienced by parents, is required before meaningful policy and practice recommendations can be made. PMID- 17125522 TI - Antibacterial resistance and their genetic location in MRSA isolated in Kuwait hospitals, 1994-2004. AB - BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) continues to be a major cause of serious infections in hospitals and in the community worldwide. In this study, MRSA isolated from patients in Kuwait hospitals were analyzed for resistance trends and the genetic location of their resistance determinants. METHODS: Between April 1994 and December 2004, 5644 MRSA isolates obtained from different clinical samples were studied for resistance to antibacterial agents according to guidelines from the National Committee for Clinical Laboratory Standards and the British Society for Antimicrobial Chemotherapy. The genetic location of their resistance determinants was determined by curing and transfer experiments. RESULTS: They were resistant to aminoglycosides, erythromycin, tetracycline, trimethoprim, fusidic acid, ciprofloxacin, chloramphenicol, rifampicin, mupirocin, cadmium acetate, mercuric chloride, propamidine isethionate and ethidium bromide but susceptible to vancomycin, teicoplanin and linezolid. The proportion of the isolates resistant to erythromycin, ciprofloxacin and fusidic acid increased during the study period. In contrast, the proportion of isolates resistant to gentamicin, tetracycline, chloramphenicol and trimethoprim declined. High-level mupirocin resistance increased rapidly from 1996 to 1999 and then declined. They contained plasmids of 1.9, 2.8, 3.0, 4.4, 27 and 38 kilobases. Genetic studies revealed that they carried plasmid-borne resistance to high-level mupirocin resistance (38 kb), chloramphenicol (2.8-4.4 kb), erythromycin (2.8-3.0 kb) and cadmium acetate, mercuric chloride, propamidine isethionate and ethidium bromide (27 kb) and chromosomal location for methicillin, the aminoglycosides, tetracycline, fusidic acid, ciprofloxacin and trimethoprim resistance. Thus, the 27 kb plasmids had resistance phenotypes similar to plasmids reported in MRSA isolates in South East Asia. CONCLUSION: The prevalence of resistance to erythromycin, ciprofloxacin, high-level mupirocin and fusidic acid increased whereas the proportion of isolates resistant to gentamicin, tetracycline, chloramphenicol and trimethoprim declined during the study period. They contained 27-kb plasmids encoding resistance to cadmium acetate, mercuric chloride, propamidine isethionate and ethidium bromide similar to plasmids isolated in MRSA from South East Asia. Molecular typing of these isolates will clarify their relationship to MRSA from South East Asia. PMID- 17125523 TI - Establishment of a pipeline to analyse non-synonymous SNPs in Bos taurus. AB - BACKGROUND: Single nucleotide polymorphisms (SNPs) are an abundant form of genetic variation in the genome of every species and are useful for gene mapping and association studies. Of particular interest are non-synonymous SNPs, which may alter protein function and phenotype. We therefore examined bovine expressed sequences for non-synonymous SNPs and validated and tested selected SNPs for their association with measured traits. RESULTS: Over 500,000 public bovine expressed sequence tagged (EST) sequences were used to search for coding SNPs (cSNPs). A total of 15,353 SNPs were detected in the transcribed sequences studied, of which 6,325 were predicted to be coding SNPs with the remaining 9,028 SNPs presumed to be in untranslated regions. Of the cSNPs detected, 2,868 were predicted to result in a change in the amino acid encoded. In order to determine the actual number of non-synonymous polymorphic SNPs we designed assays for 920 of the putative SNPs. These SNPs were then genotyped through a panel of cattle DNA pools using chip-based MALDI-TOF mass spectrometry. Of the SNPs tested, 29% were found to be polymorphic with a minor allele frequency >10%. A subset of the SNPs was genotyped through animal resources in order to look for association with age of puberty, facial eczema resistance or meat yield. Three SNPs were nominally associated with resistance to the disease facial eczema (P < 0.01). CONCLUSION: We have identified 15,353 putative SNPs in or close to bovine genes and 2,868 of these SNPs were predicted to be non-synonymous. Approximately 29% of the non synonymous SNPs were polymorphic and common with a minor allele frequency >10%. Of the SNPs detected in this study, 99% have not been previously reported. These novel SNPs will be useful for association studies or gene mapping. PMID- 17125524 TI - Knowledge of breast cancer and its early detection measures among rural women in Akinyele Local Government Area, Ibadan, Nigeria. AB - BACKGROUND: Breast cancer is the commonest cancer among women in Nigeria and globally. In Nigeria, late presentations of breast cancer cases have also been consistent for three decades. In an environment where there is no established national screening program for breast cancer, it is pertinent to assess the knowledge of breast cancer and its early detection measures. The objective of this study therefore, was to assess rural women's level of knowledge of breast cancer and its early detection measures. METHODS: The knowledge of various aspects of breast cancer; etiology, early warning signs, treatment modes and early detection measures; was assessed among women in two randomly selected health districts in Akinyele Local Government in Ibadan. The assessment was performed with the use of a self-structured validated questionnaire administered by trained interviewers to 420 women randomly selected from the two health districts. The various aspects of facts about breast cancer were scored and added together to determine respondents' level of knowledge RESULTS: The mean score of knowledge of breast cancer was 55.4 SD 5.4 (range of scores obtainable was 26 78), while the mean score for knowledge of early detection of breast cancer was 24.8 SD 2.3 (range of scores obtainable was 12-36). The leading source of information about breast cancer was "elders, neighbors and friends" and 63(15.4%) acknowledged this source, while only 18 (4.4%) respondents acknowledged health workers as source. Only 54 (13.3%) claimed to have heard about breast self- examination (BSE) however, and the leading source of information about BSE were health workers. Nine (2.2%) of respondents claimed this source. CONCLUSION: This study revealed that respondents lacked knowledge of vital issues about breast cancer and early detection measures. It also revealed that health workers were not forthcoming with information to the public thereby constituting a challenge to community health nurses and other health workers, to provide vital information to the public. PMID- 17125525 TI - Specific food intake, fat and fiber intake, and behavioral correlates of BMI among overweight and obese members of a managed care organization. AB - BACKGROUND: The study examined correlates of body mass index (BMI) in overweight and obese members of a managed care organization seeking treatment for obesity. It assessed intake of specific foods, dietary fat or fiber, and behaviors attempted to control weight. METHODS: Participants were 508 men and 1293 women who were > 18 years and had a self-reported BMI > 27.0. This paper reports analyses of baseline and 24-month follow-up data from a randomized weight-loss trial. Cross-sectional and prospective relationships between BMI and behaviors were examined with regression analyses controlling for age and education. RESULTS: At baseline, hamburger and beef consumption were associated with higher BMI for men; for women, hamburger, fried chicken, hot dog, bacon or sausage, egg, French fry, and overall fat consumption were associated with higher BMI, while eating high fiber cereal, fruit, and overall fiber intake were associated with lower BMI. Virtually all forms of weight control behavior were reported more often in heavier people. Subscribing to exercise magazines, however, was associated with lower BMI. Decreased fat intake and increased fruit/vegetable/fiber intake over the course of the study were associated with reductions in BMI at 24 months. CONCLUSION: The same behaviors that differentiate individuals with different body weight in the general population also differentiate between individuals of different body weights at the high end of the weight distribution. Educational efforts aimed at preventing weight gain and reducing obesity might benefit from focusing on specific foods known to be associated empirically with body weight and weight change over time. PMID- 17125527 TI - Nuts: nutrition and health outcomes. Preface. PMID- 17125526 TI - Identification of novel citrullinated autoantigens of synovium in rheumatoid arthritis using a proteomic approach. AB - Recently, autoantibodies to some citrullinated autoantigens have been reported to be specific for rheumatoid arthritis (RA). However, an entire profile of and autoimmunity of the citrullinated proteins have been poorly understood. To understand the profile, we examined citrullinated autoantigens by a proteomic approach and further investigated the significance of citrullination in antigenicity of one of the autoantigens. Specifically, we detected citrullinated autoantigens in synovial tissue of a patient with RA by two-dimensional electrophoresis and Western blotting by using pooled sera from five patients with RA and anti-citrulline antibodies. After identifying the detected autoantigens by mass spectrometry, we investigated the contribution of citrullination to autoantigenicity by using a recombinant protein with or without citrullination on one of the identified novel citrullinated autoantigens. As a result, we found 51 citrullinated protein spots. Thirty (58.8%) of these spots were autoantigenic. We identified 13 out of the 30 detected citrullinated autoantigenic proteins. They contained three fibrinogen derivatives and several novel citrullinated autoantigens (for example, asporin and F-actin capping protein alpha-1 subunit [CapZalpha-1]). We further analyzed the contribution of citrullination to autoantigenicity in one of the detected citrullinated autoantigens, CapZalpha-1. As a result, frequencies of autoantibodies to non-citrullinated CapZalpha-1 were 36.7% in the RA group tested, 10.7% in the osteoarthritis (OA) group, and 6.5% in healthy donors. On the other hand, those to citrullinated CapZalpha-1 were 53.3% in the RA group, 7.1% in the OA group, and 6.5% in the healthy donors. This shows that autoantigenicity of citrullinated or non-citrullinated CapZalpha-1 is relevant to RA. The antibody titers to the citrullinated CapZalpha-1 were significantly higher than those to the non-citrullinated CapZalpha-1 in 36.7% of patients; however, the other patients showed almost equal antibody titers to both citrullinated and non-citrullinated CapZalpha-1. Therefore, the autoantibodies would target citrulline-related and/or citrulline-unrelated epitope(s) of CapZalpha-1. In conclusion, we report a profile of citrullinated autoantigens for the first time. Even though citrullination is closely related to autoantigenicity, citrullination would not always produce autoantigenicity in RA. Citrullinated and non-citrullinated autoantigens/autoepitopes would have different pathological roles in RA. PMID- 17125528 TI - Consumption and portion sizes of tree nuts, peanuts and seeds in the European Prospective Investigation into Cancer and Nutrition (EPIC) cohorts from 10 European countries. AB - Tree nuts, peanuts and seeds are nutrient dense foods whose intake has been shown to be associated with reduced risk of some chronic diseases. They are regularly consumed in European diets either as whole, in spreads or from hidden sources (e.g. commercial products). However, little is known about their intake profiles or differences in consumption between European countries or geographic regions. The objective of this study was to analyse the population mean intake and average portion sizes in subjects reporting intake of nuts and seeds consumed as whole, derived from hidden sources or from spreads. Data was obtained from standardised 24-hour dietary recalls collected from 36 994 subjects in 10 different countries that are part of the European Prospective Investigation into Cancer and Nutrition (EPIC). Overall, for nuts and seeds consumed as whole, the percentage of subjects reporting intake on the day of the recall was: tree nuts = 4. 4%, peanuts = 2.3 % and seeds = 1.3 %. The data show a clear northern (Sweden: mean intake = 0.15 g/d, average portion size = 15.1 g/d) to southern (Spain: mean intake = 2.99 g/d, average portion size = 34.7 g/d) European gradient of whole tree nut intake. The three most popular tree nuts were walnuts, almonds and hazelnuts, respectively. In general, tree nuts were more widely consumed than peanuts or seeds. In subjects reporting intake, men consumed a significantly higher average portion size of tree nuts (28.5 v. 23.1 g/d, P<0.01) and peanuts (46.1 v. 35.1 g/d, P<0.01) per day than women. These data may be useful in devising research initiatives and health policy strategies based on the intake of this food group. PMID- 17125529 TI - Nuts: source of energy and macronutrients. AB - On the basis of the high fat content of nuts, they are traditionally considered as foods that provide a high amount of energy. However, epidemiologic and clinical observations do not indicate an association between nut intake and increased BMI. There is a notorious variability in macronutrient composition among nuts, although they have some consistent patterns. Nuts contain all major macronutrients: protein, carbohydrate, and fat. The total protein content is relatively high, which makes them a good source of plant protein (especially for vegetarians). Although nuts contain low amounts of some essential amino acids, this is not a nutritional concern due to the complement of protein. In addition, nuts have a low lysine:arginine ratio, which is inversely associated with the risk of developing hypercholesterolemia and atherosclerosis. Carbohydrates are the second highest macronutrient in nuts in terms of total calories provided. The fat fraction is characterized by a high amount of unsaturated fatty acids and a low content of saturated fatty acids. In conclusion, the high content in unsaturated fatty acids, the low lysine:arginine ratio, and the presence of other bioactive molecules (such as fibre, phytosterols, vitamin and other antioxidants, and minerals) make the addition of nuts to healthy diets a useful tool for the prevention of cardiovascular heart diseases. PMID- 17125530 TI - Fatty acid composition of nuts--implications for cardiovascular health. AB - It is well established that, due to their high content of saturated fatty acids (SFA), the intake of meat and meat products is strongly associated with elevated blood cholesterol concentrations and an increased risk of hypertension, diabetes and cardiovascular diseases. Conversely, the intake of foods rich in unsaturated fatty acids, such as those contained in most vegetable fats and oils and oily fish, is associated with improved lipid profiles, a lower potency of intermediate biomarkers of atherosclerosis and lesser incidence of cardiovascular diseases. There are persuasive evidences that dietary substitution of monounsaturated fatty acids (MUFA) or n-6 polyunsaturated fatty acids (PUFA) for SFA lowers blood cholesterol and may have beneficial effects on inflammation, thrombosis, and vascular reactivity. MUFA may have an advantage over PUFA because enrichment of lipoprotein lipids with MUFA increases their resistance to oxidation. Marine n-3 PUFA have a number of anti-atherosclerotic effects, including anti-arrhythmic properties and, at relatively high doses, reduce serum triglycerides. These effects appear to be shared in part by vegetable n-3 PUFA. Nuts are natural foods rich in unsaturated fatty acids; most nuts contain substantial amounts of MUFA, while walnuts are especially rich in both n-6 and n-3 PUFA. Healthy fats in nuts contribute to the beneficial effects of frequent nut intake observed in epidemiological studies (prevention of coronary heart disease, diabetes, and sudden death) and in short-term feeding trials (cholesterol lowering, LDL resistance to oxidation, and improved endothelial function). PMID- 17125531 TI - Nut consumption in Spain and other countries. AB - In countries of the Mediterranean region, nuts have been consumed in moderate quantities since ancient times. Epidemiological studies show lower risk of cardiovascular diseases in populations with frequent nut consumption, independent from other dietary components. This article assesses nut consumption in Spain and other countries using different sources of data collected at the country, household or individual levels. The per capita consumption of nuts in Spain in 2001 was 7.9 g/person/d. The varieties most widely consumed are walnuts, almonds, hazelnuts and peanuts. Results of the eVe study estimate an average nut consumption in the Spanish population aged 25-60 years of 3.3 g/person/d. No significant statistical differences were observed between men and women. Consumption is higher in men aged between 35 and 44 years (4.5 g/d) and in women aged between 45 and 54 years (3.5 g/d). In the population of 2-24 years, according to the enKid study, nut consumption is estimated at 4.9 +/- 18.5 g/person per d. The age group with the highest consumption is teenagers between 14 and 17 years. The northeastern, northern and eastern regions of Spain show the highest consumption. According to FAO balance sheets, in 2001, Lebanon (16.5 kg/person per year) and Greece (11.9 kg/person per year) were the countries in the Mediterranean region with the highest consumption of nuts, followed by Spain (7.3 kg/person per year), Israel and Italy. PMID- 17125532 TI - Other relevant components of nuts: phytosterols, folate and minerals. AB - Nuts contain significant amounts of essential micronutrients that are associated with an improved health status when consumed at doses beyond those necessary to prevent deficiency states. Nuts do not contain cholesterol, but they are rich in chemically related phytosterols, a class of compounds that interfere with intestinal cholesterol absorption and thus help lower blood cholesterol. Nuts also contain folate, a B-vitamin necessary for normal cellular function that plays an important role in detoxifying homocysteine, a sulphur-containing amino acid with atherothrombotic properties that accumulates in plasma when folate status is subnormal. Compared to other common foodstuffs, nuts have an optimal nutritional density with respect to healthy minerals, such as calcium, magnesium and potassium. Like that of most vegetables, the sodium content of nuts is very low. A high intake of calcium, magnesium and potassium, together with a low sodium intake, is associated with protection against bone demineralisation, arterial hypertension, insulin resistance, and overall cardiovascular risk. Phytosterols might justify part of the cholesterol-lowering effect of nut intake beyond that attributable to fatty acid exchange, while the mineral richness of nuts probably contributes to the prevention of diabetes and coronary heart disease observed in epidemiological studies in association with frequent nut consumption. PMID- 17125533 TI - Dietary fibre, nuts and cardiovascular diseases. AB - Dietary fibre has a range of metabolic health benefits. Through a variety of mechanisms, dietary fibre, and the viscous variety in particular, slows down gastric emptying and intestinal transit, decreases the rate of intestinal carbohydrate absorption, and increases faecal bile acid excretion. Therefore, consumption of some types of soluble fibre can enhance satiety, which is associated with a lower BMI, and reduce blood cholesterol and the postprandial glucose response. Surprisingly, the consumption of insoluble fibre from whole grains, though metabolically inert, has been associated with a reduction in the risk of developing coronary heart disease and diabetes in epidemiological studies. The likely reason is that whole grains, like nuts, legumes and other edible seeds, contain many bioactive phytochemicals and various antioxidants. After cereals, nuts are the vegetable foods that are richest in fibre, which may partly explain their benefit on the lipid profile and cardiovascular health. PMID- 17125534 TI - Health benefits of nuts: potential role of antioxidants. AB - A diet rich in fruits, vegetables and minimally refined cereals is associated with lower risk for chronic degenerative diseases. Since oxidative stress is common in chronic degenerative disease, it has been assumed that dietary antioxidants may explain this protective effect. Every dietary plant contains numerous types of antioxidants with different properties. Many of these antioxidants cooperate in oxidative stress reduction in plants, and we hypothesize that many different antioxidants may also be needed for the proper protection of animal cells. To test this hypothesis, it is useful to identify dietary plants with high total antioxidant content. Several nuts are among the dietary plants with the highest content of total antioxidants. Of the tree nuts, walnuts, pecans and chestnuts have the highest contents of antioxidants. Walnuts contain more than 20 mmol antioxidants per 100 g, mostly in the walnut pellicles. Peanuts (a legume) also contribute significantly to dietary intake of antioxidants. These data are in accordance with our present extended analysis of an earlier report on nut intake and death attributed to various diseases in the Iowa Women's Health Study. We observed that the hazard ratio for total death rates showed a U-shaped association with nut/peanut butter consumption. Hazard ratio was 0.89 (CI = 0.81-0.97) and 0.81 (CI = 0.75-0.88) for nut/peanut butter intake once per week and 1-4 times per week, respectively. Death attributed to cardiovascular and coronary heart diseases showed strong and consistent reductions with increasing nut/peanut butter consumption. Further studies are needed to clarify whether antioxidants contribute to this apparent beneficial health effect of nuts. PMID- 17125535 TI - Nuts and coronary heart disease: an epidemiological perspective. AB - The epidemiological evidence for the cardio-protective effect of nut consumption is presented and reviewed. Four large prospective epidemiological studies of primary prevention of coronary heart disease are reviewed and discussed (Adventist Health Study, Iowa Women's Health Study, Nurses' Health Study and the Physicians' Health Study). Other studies of nuts and coronary heart disease risk are addressed. The combined evidence for a cardio-protective effect from nut consumption is summarized and presented graphically. The risk of coronary heart disease is 37 % lower for those consuming nuts more than four times per week compared to those who never or seldom consume nuts, with an average reduction of 8.3 % for each weekly serving of nuts. The evidence for a causal relationship between nut consumption and reduced risk of coronary heart disease is outlined using Hill's criteria for causality and is found to support a causal cardio protective relationship. PMID- 17125536 TI - Tree nuts and the lipid profile: a review of clinical studies. AB - Tree nuts have a fatty acid profile that favourably affects blood lipids and lipoproteins. They are low in saturated fat and high in unsaturated fatty acids and are rich sources of other nutrients. An extensive database consistently shows total and LDL cholesterol-lowering effects of diets low in saturated fat and cholesterol and high in unsaturated fat provided by a variety of tree nuts. Collectively, a summary of studies conducted to date shows that tree nuts reduce LDL cholesterol by 3-19 % compared with Western and lower-fat diets. Nuts also contain many nutrients and bioactive compounds that appear to contribute to the favourable effects on lipids and lipoproteins--these include plant sterols, dietary fibre and antioxidants. Because of their unique nutrient profile, nuts can be part of a diet that features multiple heart-healthy foods resulting in a cholesterol lowering response that surpasses that of cholesterol-lowering diets typically used to reduce CVD risk. PMID- 17125537 TI - Nuts, body weight and insulin resistance. AB - Traditionally, nuts have been considered a staple food, but because of their high energy and fat content are not considered good for body weight control or insulin sensitivity. Frequent consumption of nuts reduces the risk of coronary artery disease and type-2 diabetes and nut-enriched diets favourably alter blood lipids in normal and hypercholesterolemic individuals under controlled and free-living dietary conditions. However, whether or not frequent consumption of nuts can cause weight gain and impair insulin sensitivity is not fully understood. Review of the available data to date suggests that adding nuts to habitual diets of free living individuals does not cause weight gain. In fact, nuts have a tendency to lower body weight and fat mass. In the context of calorie-restricted diets, adding nuts produces a more lasting and greater magnitude of weight loss among obese subjects while improving insulin sensitivity. Further studies are needed to clarify the effect of long-term (>/= year) consumption of nuts on body weight and their role in altering insulin sensitivity both in normal and type-2 diabetics. In the meantime, there is sufficient evidence to promote the inclusion of nuts as part of healthy diets. PMID- 17125538 TI - The potential of nuts in the prevention of cancer. AB - Cancer is a disease that is characterized by the loss of genetic control over cell growth and proliferation, mainly as a result of the exposure to environmental factors. Cessation of smoking and a high consumption of fruits and vegetables are the most important means of reducing the risk of cancer in our society. Like fruits and vegetables, nuts are a source of vegetable protein, monounsaturated fatty acids, vitamin E, phenolic compounds, selenium, vegetable fibre, folic acid and phytoestrogens. There are numerous mechanisms of action by which these components can intervene in the prevention of cancer, although they have not been fully elucidated. There are very few epidemiological studies analyzing the relationship between nuts consumption and risk of cancer. One of the greatest difficulties in interpreting the results is that the consumption of nuts, seeds and legumes are often presented together. The most commonly studied location is the colon/rectum, an organ in which the effect of nuts is biologically plausible. Although the results are not conclusive, a protective effect on colon and rectum cancer is possible. Likewise, some studies show a possible protective effect on prostate cancer, but there is insufficient data on other tumour locations. New epidemiological studies are required to clarify the possible effects of nuts on cancer, particularly prospective studies that make reliable and complete estimations of their consumption and which make it possible to analyse their effects independently of the consumption of legumes and seeds. PMID- 17125539 TI - Food allergy: nuts and tree nuts. AB - Nuts are a well-defined cause of food allergy, which affect approximately 1 % of the general population in the UK and the USA. There do appear to be differences in the frequency of nut allergy between different countries because of different dietary habits and cooking procedures. For example, in the USA and France, peanuts are one of the most frequent causes of food allergy, but in other countries, it seems to be less common. Genetic factors, in particular, appear to play a role in the development of peanut allergy. While the majority of nut allergens are seed storage proteins, other nut allergens are profilins and pathogenesis-related protein homologues, considered as panallergens because of their widespread distribution in plants. The presence of specific IgE antibodies to several nuts is a common clinical finding, but the clinical relevance of this cross-reactivity is usually limited. Allergic reactions to nuts appear to be particularly severe, sometimes even life-threatening, and fatal reactions following their ingestion have been documented. Food allergy is diagnosed by identifying an underlying immunological mechanism (i.e. allergic testing), and establishing a causal relationship between food ingestion and symptoms (i.e. oral challenges). In natural history investigations carried out in peanut-allergic children, approximately 20 % of the cases outgrew their allergy or developed oral tolerance. The treatment of nut allergies should include patient and family education about avoiding all presentations of the food and the potential for a severe reaction caused by accidental ingestion. Patients and families should be instructed how to recognise early symptoms of an allergic reaction and how to treat severe anaphylaxis promptly. PMID- 17125540 TI - The life cycle of Haplorchis pumilio (Trematoda: Heterophyidae) from the Indian region. AB - The life cycle of the heterophyid fluke, Haplorchis pumilio is elucidated for the first time from the Indian region. Various stages in the life cycle were established based on observations made on natural infections found in snails and fish in a freshwater stream at Visakhapatnam, India and experimental infections carried out in the laboratory. The thiarid snail, Thiara tuberculata served as the first intermediate host and a wide range of freshwater fish as second intermediate hosts. Natural infections with adult flukes were found in the piscivorous birds Ardeola grayii and Bubulcus ibis. Adults were raised experimentally in day-old chicks. Distinguishing features of the cercaria of H. pumilio are: a large body size (200-224 x 92-96 micro m), body-tail ratio of 1:2.1 and densely distributed pigment granules in the parenchyma imparting a brownish tinge to the body. Natural infections with metacercariae were found in the freshwater fish Channa punctatus, C. orientalis, Puntius sophore, Gambusia affinis and fingerlings of Cyprinus carpio and Liza macrolepis. Additionally, experimental infections were established in Therapon jarbua, Esomus danricus and Oreochromis mossambica. Metacercariae were embedded in the caudal muscles of fish and heavy infections induced mortality. Metacercariae were infective at about 15 days of age. PMID- 17125541 TI - Rhinoclemmysnema n. g. and three new species of nematodes of the family Atractidae (Cosmocercoidea), with notes on the helminth fauna of Rhinoclemmys pulcherrima (Testudines: Bataguridae) in Costa Rica. AB - Rhinoclemmysnema n. g. and three new species of atractid nematodes, namely, Atractis costaricaensis n. sp., Orientatractis asymmetrica n. sp. and Rhinoclemmysnema multilabiatum were recovered from the small and large intestine of Rhinoclemmys pulcherrima, the painted wood turtle in the Guanacaste Conservation Area, Costa Rica. The genera and three species are characterized by the features of the cephalic region and male tail. Neopolystoma fentoni (conjuctival sac) and Heronimus mollis (lungs) were also collected. PMID- 17125542 TI - Morphological observations and the effects of artificial digestive fluids on the survival of Diploscapter coronata from a Japanese patient. AB - Unusual non-human parasitic nematodes and eggs were detected in the faeces of an 8-year-old Japanese female suffering from Henoch-Schonlein purpura. The worms were adult female rhabditiform nematodes measuring 325.6-441.2 micro m in length and 18.3-26.5 micro m in width. One pair of the labia oris was notched with many spiny projections, while the other pair was strongly curved outwards. The worms were identified using light and scanning electron microscopy as the free-living nematode Diploscapter coronata (Cobb) based on their characteristic morphology. The patient's faeces containing worms and eggs were cultured using a filter-paper culture technique and after 7 days of culture, male as well as female worms were recovered. Worm survival time and hatchability of the eggs were examined in vitro after treatment with an artificial gastric or intestinal fluid. Although adult worms survived for less than one minute, eggs hatched after treatment with artificial gastric fluid. This suggests that eggs accidentally ingested or produced by adult D. coronata could develop in the human gastro-intestinal tract. Some morphological features of male D. coronata are also described. PMID- 17125543 TI - Where in Europe should we look for sources of the cutaneous trematode Collyriclum faba infections in migrating birds? AB - Cutaneous cysts with trematodes of Collyriclum faba have been found in birds during their spring and post-breeding migrations in the Czech Republic. During spring migrations, C. faba was found in one dunnock Prunella modularis, two European robins Erithacus rubecula, three common nightingales Luscinia megarhynchos, one song thrush Turdus philomelos and one great reed warbler Acrocephalus arundinaceus. During post-breeding migration, the same parasite was found in one garden warbler Sylvia borin, one whitehroat S. communis, three goldcrests Regulus regulus and one Eurasian treecreeper Certhia familiaris. The newly identified hosts of C. faba are dunnock, common nightingale, song thrush, great reed warbler and Eurasian treecreeper. Geographical areas of the birds' infection were identified from an analysis of reports on ringed birds of the same species, the time necessary for the development of cutaneous cysts with C. faba and the time of their survival, and hitherto known geographical areas of endemic occurrence of C. faba. It is presumed that birds trapped during spring migrations were infected in some montane and submontane regions in south-western Europe (the Alps, the Apennines). Birds infected during autumn migration or post-breeding vagrancy could have been infected in the Central European Carpathians, the region of C. faba endemic occurrence. For migrating birds, the impact of C. faba infections has not been hitherto assessed. PMID- 17125544 TI - Morphological variability within Oesophagostomum bifurcum among different primate species from Ghana. AB - Adult Oesophagostomum bifurcum (Nematoda: Strongylida) from human and non-human primates from Ghana were compared in order to investigate the extent of morphological variability within the species. Using analysis of variance and principal component analysis, significant differences in morphological characters (such as parasite length, width, length of the oesophagus and length of spicules) were demonstrated between O. bifurcum worms from humans, the Mona, Patas or Green monkey and/or Olive baboons. These findings suggest that O. bifurcum from different species of primate host represent distinct population variants, also supported by recent epidemiological and genetic studies of O. bifurcum from such hosts. PMID- 17125545 TI - Diagnostic value of IgG isotype responses against Brugia malayi antifilarial antibodies in the clinical spectrum of brugian filariasis. AB - To study the diagnostic significance of antifilarial IgG subclasses in the clinical spectrum of brugian filariasis, IgG1, IgG3 and IgG4 antifilarial antibodies were determined in an exposed population comprising 74 asymptomatic amicrofilaraemics, 30 microfilaraemics, 20 lymphangitis and 16 elephantiasis patients resident in Narathiwart province, an area endemic for Brugia malayi lymphatic filariasis in southern Thailand. The dominant isotype of antifilarial antibody was IgG4. A significantly higher percentage of individuals were positive for IgG1 in the microfilaraemic and lymphangitis groups compared with the elephantiasis and endemic normal patients, while a significantly higher positive rate of IgG3 was found in those with lymphangitis. The possible role of these isotypes for diagnostic purposes and the pattern of antibody response in various clinically manifesting groups are discussed. PMID- 17125546 TI - Helminth communities of Nectomys squamipes naturally infected by the exotic trematode Schistosoma mansoni in southeastern Brazil. AB - The water rat Nectomys squamipes is endemic in Brazil and found naturally infected with Schistosoma mansoni. Helminth communities, their prevalences, intensity of infection and abundance in N. squamipes in an endemic area of schistosomiasis in the state of Rio de Janeiro, Brazil were studied. Four species of nematodes (Physaloptera bispiculata, Syphacia venteli, Hassalstrongylus epsilon and Litomosoides chagasfilhoi) were recovered in 85.3%, two trematodes (Schistosoma mansoni and Echinostoma paraensei) in 38.8% and one cestode species (Raillietina sp.) in 1.7% of rats examined. Rats were infected with up to five helminth species each, and these were highly aggregated in distribution. For H. epsilon and S. venteli, intensities and abundances were higher in adult male and subadult female hosts, respectively. Hassaltrongylus epsilon, P. bispiculata, S. venteli and S. mansoni were classified as dominant species, L. chagasfilhoi and E. paraensei as co-dominant and Raillietina sp. as subordinated. No significant correlation was found in the intensity of infecton between each pair of helminth species. Schistosoma mansoni was not related to any other helminth species according to their infection rates, althougth S. mansoni was well established in the natural helminth comunity of the water rat. PMID- 17125547 TI - The ultrastructure of hypersymbionts on the monogenean Gyrodactylus salaris infecting Atlantic salmon Salmo salar. AB - There is increasing pressure to develop alternative control strategies against the pathogen Gyrodactylus salaris, which has devastated wild Atlantic salmon Salmo salar in Norway. Hyperparasitism is one option for biological control and electron microscopy has revealed two ectosymbionts associated with G. salaris: unidentified rod-shaped bacteria, and the protist, Ichthyobodo necator. No endosymbionts were detected. The flagellate I. necator occurred only occasionally on fish suffering costiosis, whereas bacterial infections on the tegument of G. salaris were observed throughout the year, but at variable densities. Bacteria were seldom observed attached to fish epidermis, even when individuals of G. salaris on the same host were heavily infected. Wounds on salmon epidermis caused by the feeding activity of bacteria-infected G. salaris did not appear to be infected with bacteria. On heavily infected gyrodactylids, bacteria were most abundant anteriorly on the cephalic lobes, including the sensory structures, but no damaged tissue was detected by transmission electron microscopy in the region of bacterial adherence. Furthermore, transmission and survival of infected G. salaris on wild salmon did not appear to be influenced by the bacterial infection. The lack of structural damage and impact on G. salaris biology indicates that these bacteria are not a potential agent for control of gyrodactylosis. However, this may not be the case for all gyrodactylid-bacterial interactions and a review of bacterial infections of platyhelminths is presented. PMID- 17125548 TI - Serodiagnostic potential of chemically synthesized glycosphingolipid antigens in an enzyme-linked immunosorbent assay for alveolar echinococcosis. AB - In the serodiagnosis of alveolar echinococcosis, the detection of specific reactions against not only protein but also carbohydrate antigen is useful and both antigens supplement each other. Though recombinant protein antigens have recently advanced, the preparation of carbohydrate antigen still depends on extraction from crude antigens. In the latter case, it is not conventional to obtain carbohydrate antigen as a single component for examination and research. Therefore, chemically synthesized carbohydrate antigens were prepared for serodiagnosis by the enzyme-linked immunosorbent assay (ELISA). Four antigens with the structure of glycosphingolipids from Echinococcus multilocularis were examined and one antigen, Galbeta1-6(Fucalpha1-3)Galbeta1-6Galbeta1-ceramide, was found to show significant serodiagnostic potential in differentiating alveolar from cystic echinococcosis. PMID- 17125549 TI - Evaluation of broad-spectrum anthelmintic activity in a novel assay against Haemonchus contortus, Trichostrongylus colubriformis and T. sigmodontis in the gerbil Meriones unguiculatus. AB - The gerbil Meriones unguiculatus, infected with three species of nematodes, each located in a separate part of the gastrointestinal tract, provided a reliable laboratory assay for the evaluation of broad-spectrum anthelmintic activity. Gerbils harbouring 6-day-old infections of Haemonchus contortus, Trichostrongylus colubriformis and T. sigmodontis were given selected broad-spectrum anthelmintics by gavage. Three benzimidazoles, thiabendazole, oxfendazole and albendazole, a tetrahydropyrimidine, morantel, an imidazothiazole, levamisole hydrochloride, a macrocyclic lactone, ivermectin and an experimental natural product, paraherquamide, were active against all three nematodes at various dosages. Trichostrongylus colubriformis was most sensitive to levamisole hydrochloride, morantel, thiabendazole and paraherquamide whereas ivermectin, oxfendazole and albendazole were more effective against H. contortus. All compounds were active against the caecal nematode T. sigmodontis although it was less sensitive than T. colubriformis. Haemonchus contortus was more sensitive than T. sigmodontis to all anthelmintics tested except thiabendazole. PMID- 17125550 TI - The effect of the hookworm Ancylostoma ceylanicum on the mucosal architecture of the small intestine in hamsters. AB - Hookworms are known to cause marked changes to the intestinal mucosa, especially in relation to erosion of the villi. However, since the development of enteropathy has not been examined thoroughly through quantitative experiments on infected animals, the results of experiments conducted in hamsters infected with Ancylostoma ceylanicum are reported. Changes to intestinal architecture were first apparent between 12 and 14 days after infection, and then increased in intensity for 3-4 weeks, persisting for as long as worms were present (>63 days). Following infection, the height of villi declined from a mean of 1002 micro m in naive controls to less than 200 micro m and as low as 18 micro m in one case. The depth of the crypts of Lieberkuhn increased from a baseline value of 166 micro m in naive controls to in excess of 600 micro m within 6 weeks of infection. Mitotic figures had a baseline value of 5.5 per villus-crypt unit, and this rose to in excess of 25 in some experiments. Changes were dependent on the intensity of the parasite burden on day 20, but by 30 days after infection changes in all three values were maximal and density-dependent relationships were no longer clearly apparent. Villus height and crypt depth returned to near normal values within a week of the removal of worms, although group means for both remained different from naive controls for at least 3 weeks after treatment. Cellular division, as reflected in numbers of mitotic figures, stayed elevated for over 5 weeks after removal of worms. The results suggest that enteropathy in hookworm infections stems from a combination of intestinal immune responses and from the grazing activities of the adult worms on the mucosal surface, but is not sufficient per se for expulsion of this parasite. PMID- 17125551 TI - The effect of an indirect anthelmintic treatment on parasites and breeding success of free-living pheasants Phasianus colchicus. AB - In Great Britain free-living common pheasants Phasianus colchicus are often managed at high densities owing to their popularity as a quarry species. They are prone to infection by a range of parasite species including Heterakis gallinarum, Capillaria spp. and Syngamus trachea. In 1995 the efficacy of an indirect anthelmintic technique for controlling parasitic worm burdens of pheasants was determined in a pilot study on a shooting estate in the south of England. Between 2000 and 2003 a large-scale field experiment was conducted on nine estates in eastern England to determine the effect of the technique on parasite burden and pheasant breeding success. In the absence of anthelmintic treatment worm burdens increased rapidly through March and April, whereas birds given anthelmintic treated grain had lower worm burdens during the same period. The breeding success of pheasants was significantly higher on plots provided with anthelmintic treatment, although no long-term increases in population densities were observed. The burdens of the most common parasite H. gallinarum were significantly lower in pheasants from treatment plots six weeks after the anthelmintic treatment had ceased, but spring treatment did not influence parasite burden in the following winter. PMID- 17125552 TI - Eosinophil cationic protein, specific IgE and IgG4 in human toxocariasis. AB - Among 67 French patients presenting a toxocaral infection, various demographic, environmental, clinical and laboratory parameters (blood eosinophil count, eosinophil cationic protein (ECP), serum total IgE, specific IgE against common inhalant allergens, specific IgE and IgG4 against Toxocara excretory-secretory antigens) were investigated. Correlation studies and logistic regression analyses were conducted, testing elevated levels of ECP, specific anti-Toxocara IgE or IgG4 as outcome variables An elevated ECP level was significantly associated with both cough and rhinitis, a high level of specific anti-Toxocara IgE with itchy rashes and possible atopic status, and an increase of specific anti-Toxocara IgG4 with rural residence. PMID- 17125553 TI - Human Gongylonema infection in Iran. AB - The first human infection with Gongylonema in Iran is reported in a 35-year-old Iranian woman with complaints of one year duration and treated as a psychotic patient. Two worms, a male, and a female, were retrieved, described, and identified as G. pulchrum based on their morphological characteristics. PMID- 17125555 TI - Out of the Christmas box. PMID- 17125556 TI - Vitamin A status among children in China. AB - OBJECTIVE: The objective of this study was to assess the prevalence of vitamin A deficiency (VAD) in children under 6 years old in China and to identify risk groups for VAD. DESIGN: A cross-sectional survey was conducted in 14 provinces from coastal, inland and western geographic areas in China. SETTING: One city (urban) and two counties (rural) were randomly selected from each province as survey areas. SUBJECTS: About 200 children aged 0-6 years were randomly selected in each survey area. A blood sample was collected from each child. Data on sociodemographics and nutrition were obtained by interview of the mother or principal caregiver. Fluorescence microanalysis was used to analyse serum retinol concentration. RESULTS: VAD (serum retinol <0.7 micro mol l(-1)) was observed in 957 out of 7,826 children aged 0-6 years (12.2% of the entire study population), whereas severe VAD (serum retinol <0.35 micro mol l(-1)) was found in 39 children (0.5%). The highest prevalences of VAD at >1 year of age were observed among children of mothers with minority ethnicity (22.7%) or poor education (19.8%) and in the poor western area (17.4%). CONCLUSIONS: VAD is a nutritional problem in children in China. Children living in the poor western area, having a mother with minority ethnicity or a mother with poor education have a high risk of VAD. PMID- 17125557 TI - Studies on the nutritional status of children aged 0-5 years in a drought affected desert area of western Rajasthan, India. AB - OBJECTIVE: The present study was undertaken to assess the impact of drought on the nutritional status of pre-school children aged 0-5 years from a rural population in a desert area facing drought conditions very frequently. DESIGN: The sampling design for assessment was the three-stage sampling technique. SETTING: The study was carried out in 24 villages belonging to six tehsils (sub units of district) of Jodhpur District, a drought-affected desert district of western Rajasthan, during a drought in 2003. SUBJECTS: A total of 914 children were examined at household level, with nutritional status assessed by anthropometry, dietary intake and clinical signs of nutritional deficiency. RESULTS: The results revealed growth retardation. Stunting (malnutrition of long duration) was observed in 53% of children and underweight in 60%. Wasting, an indicator of short-duration malnutrition, was present in 28% of children. The extent of malnutrition was significantly higher in girls than boys (P<0.05). Vitamin A and B complex deficiencies were found in 0.7 and 3.0% of children, respectively. Prevalence of marasmus (protein-energy malnutrition, PEM) was 1.7% (2.3% in boys and 1.1% in girls). Overall deficits in mean energy and protein intakes were very high (76 and 54%, respectively). Comparison of the present drought results with earlier studies in desert normal and desert drought conditions showed higher prevalence of PEM and higher dietary energy and protein deficiencies. CONCLUSIONS: The prevalence of wasting was high, greater than the cut-off point of 15% stated by the World Health Organization to indicate that the severity of malnutrition is critical. PEM, vitamin A and B complex deficiencies and anaemia, along with dietary deficits of energy and protein, were observed to be higher than in non-desert areas. This may be due to the harsh environmental conditions in desert areas where drought occurs quite frequently and adversely affects the economy, largely by eroding the coping capacity and economic potential of the people as a result of heavy livestock losses and reduced harvests, leading to increased poverty and poor food intake of the inhabitants. Due to inadequate consumption of daily food the children were suffering from wasting and PEM. Efforts should be made to incorporate measures, such as ensuring the supply of adequate energy and protein to all age groups and especially pre school children, into ongoing nutrition programmes in order to improve the food security of local inhabitants in this area. PMID- 17125558 TI - Mini Nutritional Assessment of rural elderly people in Bangladesh: the impact of demographic, socio-economic and health factors. AB - OBJECTIVE: In stating the Millennium Development Goals, the United Nations aims to halve malnutrition around the world by 2015. Nutritional status of the elderly population in low-income countries is seldom focused upon. The present study aimed to evaluate the magnitude of malnutrition among an elderly population in rural Bangladesh. DESIGN AND SETTING: Data collection for a multidimensional cross-sectional study of community-based elderly people aged 60 years and over was conducted in a rural area in Bangladesh. SUBJECTS: Of 850 randomly selected elderly individuals, 625 participated in home interviews. Complete nutritional information was available for 457 individuals (mean age 69+/-8 years, 55% female). Nutritional status was assessed using an adapted form of the Mini Nutritional Assessment (MNA) including body mass index (BMI). Age, sex, education, household expenditure on food and self-reported health problems were investigated as potential predictors of nutritional status. RESULTS: BMI<18.5 kg m(-2), indicating chronic energy deficiency, was found in 50% of the population. MNA revealed a prevalence of 26% for protein-energy malnutrition and 62% for risk of malnutrition. Health problems rather than age had a negative impact on nutritional status. Level of education and food expenditure were directly associated with nutritional status. CONCLUSION: In order to reduce world hunger by half in the coming decade, it is important to recognise that a substantial proportion of the elderly population, particularly in low-income countries, is undernourished. PMID- 17125559 TI - Knowledge of dietary and behaviour-related determinants of non-communicable disease in urban Senegalese women. AB - OBJECTIVE: To assess knowledge of dietary and behaviour-related determinants of non-communicable disease (NCD) of urban Senegalese women. DESIGN: A cross sectional, population study using an interviewer-administered knowledge questionnaire, developed and validated for this study. The questionnaire consisted of 24 items with six scores measuring knowledge of: (1) diet- and behaviour-related causes of NCD; (2) diet quality-NCD relationship; (3) fruit and vegetable link with NCD; (4) health consequences of obesity; (5) causes of cardiovascular disease (CVD); and (6) causes of certain cancers. SUBJECTS: A random sample of 301 women aged 20-50 years. RESULTS: The knowledge scores developed suggest that the health consequences of obesity (mean score of 65.4%) were best understood followed by causes of CVD (mean score of 60.6%), because obesity, smoking, high blood cholesterol and dietary fat were well recognised as risk factors for CVD. Subjects scored least for their knowledge of the protective effect of fruit and vegetables (mean score of 19.9%). Knowledge of causes of certain cancers (mean score of 36.1%) was also low. Women who worked outside the home had better knowledge for two scores but otherwise no relationship was found between knowledge and literacy, formal education or body mass index. CONCLUSIONS: Findings suggest reasonable overall knowledge concerning diet and behaviour with NCD, especially given the relatively new context of the obesity epidemic in Senegal. However, there was poor knowledge of the benefit of eating fruit and vegetables and other preventable causes of certain cancers. Education targeting the benefits of vegetables and fruit may have the greatest impact on NCD prevention. PMID- 17125560 TI - Determinants of nutrition improvement in a large-scale urban project: a follow-up study of children participating in the Senegal Community Nutrition Project. AB - OBJECTIVE: To study individual determinants of differential benefit from the Senegal Community Nutrition Project (CNP) by monitoring improvement in children's weight-for-age index (WA) or underweight status (WA < -2 Z-scores) during participation. DESIGN: A follow-up study using the CNP child monitoring data. Linear general models compared variations in WA according to 14 factors describing the beneficiaries and CNP services. SETTING: Poor neighbourhoods of Diourbel, a large city in Senegal, West Africa. Over a 6-month period, the CNP provided underweight or nutritionally at-risk 6-35-month-old children with monthly growth monitoring and promotion and weekly food supplementation, provided that mothers attended weekly nutrition education sessions. SUBJECTS: All the children who participated in the first two years of the project (n=4084). RESULTS: Mean WA varied from -2.13 (standard deviation (SD) 0.82) to -1.58 (SD 0.81) Z-scores between recruitment and the end of the follow-up. The lower the child's initial WA, the greater was their increase in WA but the lower was the probability of recovery from underweight. Only 61% of underweight children recovered. Six months of CNP services may not be sufficient for catch-up growth of severely underweight children. The number of food supplement rations received was not a direct indicator of the probability of recovery. After adjustment for services received and initial WA, probability of recovery was lower in girls, in younger children, in twins and when mothers belonged to a specific ethnic group. CONCLUSIONS: Determinants of benefit from CNP differed from the risk factors for underweight. Identification of participants with a lower probability of recovery can help improve outcome. Moreover, an explanation for the lack of recovery could be that many underweight children are stunted but not necessarily wasted. PMID- 17125561 TI - Evaluation of the Food and Agriculture Organization's global school-based nutrition education initiative, Feeding Minds, Fighting Hunger (FMFH), in schools of Hyderabad, India. AB - OBJECTIVE: To assess the efficacy of the Food and Agriculture Organization's global school-based nutrition education initiative, Feeding Minds, Fighting Hunger (FMFH), in improving nutrition-related knowledge levels of schoolchildren. DESIGN: Adopting the cluster randomisation technique, five schools each in experimental and control groups were randomly chosen from the member schools of a voluntary organisation. Repeated measures of knowledge levels were carried out at three points in time to assess pre-, post-intervention and retention of knowledge gained during the intervention. Children from experimental schools were given nutrition education by teachers in the classroom setting using FMFH material. SUBJECTS/SETTINGS: Schoolchildren (n=358 in the control group and n=312 in the experimental group) of grades VIII and IX from schools in Hyderabad, India. RESULTS: The classroom-based intervention resulted in a significant improvement (P<0.01) in nutrition knowledge levels of schoolchildren in the experimental group. Significant improvement in knowledge was also observed in the control group. The effect size indicated that the improvement in knowledge levels of schoolchildren in experimental schools over control schools was medium (d=0.40), indicating the efficacy of the FMFH programme in improving nutrition-related knowledge. No significant decrease (P>0.05) in knowledge levels was observed after 2 months, indicating retention of the knowledge acquired through the intervention. CONCLUSION: The FMFH programme provides an opportunity for schoolchildren to learn more about nutrition through their teachers in a classroom setting if the lesson plans are adapted to the local circumstances. Furthermore, it has the potential to make nutrition education interactive, effective and sustainable. PMID- 17125562 TI - Prevalence of obesity, food choices and socio-economic status: a cross-sectional study in the north-west of Iran. AB - AIM: To document the epidemiological features and influencing factors of obesity in the north-west of Iran, to provide baseline information for setting up a regional population-based centre to control and prevent obesity-related disorders in the area. METHODS: In this cross-sectional study, a total of 300 subjects were selected/studied in Tabriz, one of the major cities in Iran. Data on basic characteristics, anthropometric measurements, dietary assessment and physical activity were collected. Obesity was defined as body mass index > or =30 kg m(-2) for both women and men. RESULTS: Total prevalence of obesity in the area was 22.4% (95% confidence interval (CI): 18.0-27.6). The prevalence of obesity was 24% (95% CI: 18.5-31.4) for women and 18% (95% CI: 12.5-25.6) for men. For both women and men obesity prevalence showed a positive association with age (P<0.001), while there was a negative correlation of obesity with education and income (P<0.001). Fruit consumption decreased the risk of obesity in both women and men (odds ratio (OR)=0.60, 95% CI: 0.49-0.71 vs. OR=0.62, 95% CI: 0.51-0.74, respectively). The same significant pattern was observed for the consumption of green vegetables (OR=0.71, 95% CI: 0.57-0.63 vs. OR=0.86, 95% CI: 0.77-0.98 for women and men, respectively), legumes (OR=0.70, 95% CI: 0.59-0.84 vs. OR=0.78, 95% CI: 0.66-0.91 for women and men, respectively) and dairy products (OR=0.73, 95% CI: 0.61-0.91 vs. OR=0.77, 95% CI: 0.63-0.93 for women and men, respectively). CONCLUSIONS: Our study showed that educational attainment, higher income and consumption of certain food groups (i.e. vegetables, fruits, legumes and dairy products) may decrease the risk of obesity. Our findings also indicate the crucial necessity of establishing a population-based centre for obesity in the area. The essential information is now achieved to propose to local health authorities to act accordingly. However, more population-based investigations on dietary choices are needed to develop effective preventive strategies to control overweight and obesity disorders in different regions. PMID- 17125563 TI - Interaction of two public health problems in Turkish schoolchildren: nutritional deficiencies and goitre. AB - OBJECTIVE: The aim of the present study was to determine the interaction of and association between frequency of goitre detected by palpation, nutritional status evaluated by anthropometric indices and socio-economic status in school-aged children. SUBJECTS: One thousand and eighteen prepubertal and pubertal children (aged 6-14 years) attending primary schools in an urban area were included in this study. DESIGN AND SETTING: All subjects were evaluated for the presence of goitre and nutritional status. Thyroid size was assessed using the World Health Organization's (WHO) palpation system (1960). Severity of protein-energy malnutrition (PEM) was based on WHO criteria. Children were grouped into four categories of socio-economic status. RESULTS: Eight per cent of children were detected to have goitre by palpation. Body mass index and weight-for-height were significantly lower in children who had palpable goitre than in children who did not have goitre (P<0.05). Frequencies of having palpable goitre and being stunted and underweight were especially higher in children with very low socio-economic status (P=0.016, 0.01 and 0.01, respectively). Frequency of being stunted, underweight and wasted in children with palpable goitre did not change significantly according to socio-economic status (P>0.05). In logistic regression analyses, the most important factor in detection of palpable goitre was socio economic status (B=0.517, P=0.004). Fathers' education and occupation were found to be most significant (P=0.031 and 0.020, respectively). CONCLUSION: Children detected to have palpable goitre were thinner. However, nutritional disorders were not more frequent among children with palpable goitre compared with children without goitre. Goitre and nutritional deficiencies were more common in children with lower socio-economic status but the frequency of nutritional disorders in children with palpable goitre did not change according to socio-economic status. PMID- 17125564 TI - The targeting of nutritionally at-risk children attending a primary health care facility in the Western Cape Province of South Africa. AB - AIM: The aim of this study was to determine the practices of primary health care (PHC) nurses in targeting nutritionally at-risk infants and children for intervention at a PHC facility in a peri-urban area of the Western Cape Province of South Africa. METHODOLOGY: Nutritional risk status of infants and children <6 years of age was based on criteria specified in standardised nutrition case management guidelines developed for PHC facilities in the province. Children were identified as being nutritionally at-risk if their weight was below the 3rd centile, their birth weight was less than 2500 g, and their growth curve showed flattening or dropping off for at least two consecutive monthly visits. The study assessed the practices of nurses in identifying children who were nutritionally at-risk and the entry of these children into the food supplementation programme (formerly the Protein-Energy Malnutrition Scheme) of the health facility. Structured interviews were conducted with nurses to determine their knowledge of the case management guidelines; interviews were also conducted with caregivers to determine their sociodemographic status. RESULTS: One hundred and thirty-four children were enrolled in the study. The mean age of their caregivers was 29.5 (standard deviation 7.5) years and only 47 (38%) were married. Of the caregivers, 77% were unemployed, 46% had poor household food security and 40% were financially dependent on non-family members. Significantly more children were nutritionally at-risk if the caregiver was unemployed (54%) compared with employed (32%) (P=0.04) and when there was household food insecurity (63%) compared with household food security (37%) (P<0.004). Significantly more children were found not to be nutritionally at-risk if the caregiver was financially self-supporting or supported by their partners (61%) compared with those who were financially dependent on non-family members (35%) (P=0.003). The weight results of the nurses and the researcher differed significantly (P<0.001), which was largely due to the different scales used and weighing methods. The researcher's weight measurements were consistently higher than the nurses' (P<0.00). The researcher identified 67 (50%) infants and children as being nutritionally at-risk compared with 14 (10%) by the nurses. The nurses' poor detection and targeting of nutritionally at-risk children were largely a result of failure to plot weights on the weight-for-age chart (55%) and poor utilisation of the Road to Health Chart. CONCLUSIONS: Problems identified in the practices of PHC nurses must be addressed in targeting children at nutritional risk so that appropriate intervention and support can be provided. More attention must be given to socio-economic criteria in identifying children who are nutritionally at risk to ensure their access to adequate social security networks. PMID- 17125565 TI - Indigenous peoples' food systems for health: finding interventions that work. AB - This is a short report of a 'safari' held in conjunction with the International Congress of Nutrition in September 2005, in Futululu, St. Lucia, South Africa. Participants were several members of the International Union of Nutritional Sciences Task Force on Indigenous Peoples' Food Systems and Nutrition, other interested scientists and members of the Kwa Zulu indigenous community. The paper describes the rationale for and contributions towards understanding what might be successful interventions that would resonate among indigenous communities in many areas of the world. A summary of possible evaluation strategies of such interventions is also given. PMID- 17125566 TI - Meal patterns and cooking practices in Southern France and Central England. AB - OBJECTIVE: To evaluate whether meal patterns and cooking practices in Central England and Mediterranean France conform to popular stereotypes, eating together as a household, preparation of meals, food purchasing patterns, cooking practices and eating out were investigated. DESIGN: Cross-sectional studies conducted simultaneously in April 2001 using self-administered postal questionnaires. SETTING: England (Nottingham, East Midlands) and France (Montpellier, Languedoc Roussillon). SUBJECTS: A stratified random sample of 1000 males and 1000 females aged 18-65 years was generated from the electoral roll in each country. The final sample comprised 826 subjects in England (58% males, 42% females; mean age 44 years) and 766 subjects in France (42% males, 58% females; mean age 42 years). Analyses were conducted on samples standardised for sociodemographic differences. RESULTS: The French cooked from raw ingredients most often (P<0.001), ate together as a household more regularly (P<0.001) and were most likely to follow a regular meal pattern of three meals a day. On the other hand, the English relied more on ready-prepared (P<0.001) and take-away (P<0.001) meals, as well as on energy-dense snack foods such as crisps (P<0.001). Females in both countries reported having most responsibility for preparing meals. CONCLUSIONS: Some of the study's findings confirm popular stereotypes of French and English food cultures, as the importance of the convivial aspects of eating, as well as more traditional practices such as cooking meals from basic ingredients, structured mealtimes and less between-meal snacking, remain more prominent within the French population. This may contribute to the differences in prevalence of obesity seen between the two countries. PMID- 17125567 TI - Tracking of energy and nutrient intakes from adolescence to young adulthood: the experiences of the Young Hearts Project, Northern Ireland. AB - OBJECTIVE: To assess tracking of energy and nutrient intakes between adolescence and young adulthood. DESIGN: Longitudinal study of a random sample of adolescents (aged 15 years at baseline). The extent of tracking of dietary intakes (assessed by diet history) was investigated using weighted kappa statistics (kappa). SETTING: Northern Ireland population survey. SUBJECTS: Adolescents who participated in the Young Hearts Project, Northern Ireland at age 15 years, and subsequently at young adulthood aged between 20 and 25 years (n=245 males, n=231 females). RESULTS: Despite overall increases in height and weight (both P<0.001), increases in body mass index in males (P<0.001) and body fatness in females (P<0.001), median reported intakes of energy (kJ kg(-1) day(-1)), carbohydrate (g day(-1)) and fat (g day(-1)) decreased (all P<0.001) over time. Expressed as nutrient densities (per MJ), diets at young adulthood were overall richer in thiamin, vitamin B6, total folate (all P<0.001), vitamin C (P<0.01) and vitamin D (P<0.05). Whereas the nutrient density of the males' diets decreased over time for calcium (P<0.05) and vitamin A (P<0.001), iron and riboflavin densities increased in the females' diet (P<0.001). Tracking of energy (MJ day(-1)) and nutrient intakes (expressed per MJ day(-1)) at the individual level was only poor to fair (all kappa<0.25), indicating substantial drift of subjects between the low, medium and high classes of intake with increasing age. CONCLUSIONS: These data suggest that individual dietary patterns exhibited at 15 years of age are unlikely to be predictive of dietary intakes at young adulthood. PMID- 17125568 TI - Consumption of 'extra' foods (energy-dense, nutrient-poor) among children aged 16 24 months from western Sydney, Australia. AB - OBJECTIVE: This study evaluates the contribution of energy-dense, nutrient-poor 'extra' foods to the diets of 16-24-month-old children from western Sydney, Australia. DESIGN: An analysis of cross-sectional data collected on participants in the Childhood Asthma Prevention Study (CAPS), a randomised trial investigating the primary prevention of asthma from birth to 5 years. We collected 3-day weighed food records, calculated nutrient intakes, classified recorded foods into major food groups, and further classified foods as either 'core' or 'extras' according to the Australian Guide to Healthy Eating. SETTING: Pregnant women, whose unborn child was at risk of developing asthma because of a family history, were recruited from all six hospitals in western Sydney, Australia. Data for this study were collected in clinic visits and at participants' homes at the 18-month assessment. PARTICIPANTS: Four hundred and twenty-nine children participating in the CAPS study; 80% of the total cohort. RESULTS: The mean consumption of 'extra' foods was approximately 150 g day(-1) and contributed 25-30% of the total energy, fat, carbohydrate and sodium to the diets of the study children. 'Extra' foods also contributed around 20% of fibre, 10% of protein and zinc, and about 5% of calcium. Children in the highest quintile of 'extra' foods intake had a slightly higher but not significantly different intake of energy from those in the lowest quintile. However, significant differences were evident for the percentage of energy provided by carbohydrate and sugars (higher) and protein and saturated fat (lower). The intake of most micronutrients was also significantly lower among children in the highest quintile of consumption. The intake of 'extra' foods was inversely associated with the intake of core foods. CONCLUSIONS: The high percentage of energy contributed by 'extra' foods and their negative association with nutrient density emphasise the need for dietary guidance for parents of children aged 1-2 years. These preliminary data on commonly consumed 'extra' foods and portion sizes may inform age-specific dietary assessment methods. PMID- 17125569 TI - Age-related variations in flavonoid intake and sources in the Australian population. AB - OBJECTIVE: To estimate flavonoid intake in the Australian population. DESIGN: Flavonoid consumption was estimated from 24-hour recall data and apparent consumption data using US Department of Agriculture flavonoid composition data. SUBJECTS: The National Nutrition Survey 1995 assessed dietary intake (24-hour recall) in a representative sample (n=13,858) of the Australian population aged 2 years and over. RESULTS: Analysis of the 24-hour recall data indicated an average adult intake (>18 years) of 454 mg day(-1) (92% being flavan-3-ols). Apple was the highest quercetin source until age 16-18 years, after which onion became an increasingly important prominent source. Variations in hesperetin consumption reflected orange intake. Apple, apricot and grapes were the major sources of epicatechin and catechin for children, but subsided as wine consumption increased in adulthood. Wine was the main source of malvidin. Naringenin intake remained static as a percentage of total flavonoid intake until age 19-24 years, corresponding to orange intake, and then increased with age from 19-24 years, corresponding to grapefruit intake. Apparent dietary flavonoid consumption was 351 mg person(-1) day(-1), of which 75% were flavan-3-ols. Black tea was the major flavonoid source (predominantly flavan-3-ols) representing 70% of total intake. Hesperetin and naringenin were the next most highly consumed flavonoids, reflecting orange intake. Both 24-hour recall and apparent consumption data indicated that apigenin intake was markedly higher in Australia than reported in either the USA or Denmark, presumably due to differences in consumption data for leaf and stalk vegetables and parsley. CONCLUSIONS: Tea was the major dietary flavonoid source in Australia. Flavonoid consumption profiles and flavonoid sources varied according to age. More consistent methodologies, survey tools validated for specific flavonoid intakes and enhanced local flavonoid content data for foods would facilitate better international comparisons of flavonoid intake. PMID- 17125570 TI - Evaluation of a brief pilot nutrition and exercise intervention for the prevention of weight gain in general practice patients. AB - OBJECTIVE: To pilot-test a brief written prescription recommending lifestyle changes delivered by general practitioners (GPs) to their patients. DESIGN: The Active Nutrition Script (ANS) included five nutrition messages and personalised exercise advice for a healthy lifestyle and/or the prevention of weight gain. GPs were asked to administer 10 scripts over 4 weeks to 10 adult patients with a body mass index (BMI) of between 23 and 30 kg m(-2). Information recorded on the script consisted of patients' weight, height, waist circumference, gender and date of birth, type and frequency of physical activity prescribed, and the selected nutrition messages. GPs also recorded reasons for administering the script. Interviews recorded GPs views on using the script. SETTING: General practices located across greater Melbourne. SUBJECTS AND RESULTS: Nineteen GPs (63% female) provided a median of nine scripts over 4 weeks. Scripts were administered to 145 patients (mean age: 54+/-13.2 years, mean BMI: 31.7+/-6.3 kg m(-2); 57% female), 52% of whom were classified as obese (BMI >30 kg m(-2)). GPs cited 'weight reduction' as a reason for writing the script for 78% of patients. All interviewed GPs (90%, n=17) indicated that the messages were clear and simple to deliver. CONCLUSIONS: GPs found the ANS provided clear nutrition messages that were simple to deliver. However, GPs administered the script to obese patients for weight loss rather than to prevent weight gain among the target group. This has important implications for future health promotion interventions designed for general practice. PMID- 17125571 TI - ENT surgery, blood and Jehovah's Witnesses. AB - Surgical procedures in otolaryngology are often associated with the need for blood transfusions. Homologous blood transfusions carry risks and may be unacceptable to some patient groups. The Jehovah's Witness Society is known to many because of its stance on blood products. Refusal of potentially life-saving treatment creates ethical dilemmas for treating clinicians. Throughout the world, Jehovah's Witnesses have fought for the right to refuse blood products. This article examines the need for blood in otolaryngological procedures, surgical strategies to reduce blood loss, the beliefs of Jehovah's Witnesses regarding the acceptability of blood, and procedures and legal stances adopted when treating Jehovah's Witnesses. PMID- 17125572 TI - One size fits all? A novel application of the adult Merocel epistaxis pack. PMID- 17125573 TI - Sarcoidosis of the external ear--literature review and report of a case. AB - External ear manifestations of sarcoidosis are rare. We review six cases in the literature and also report a case. The otolaryngologist plays an important role in making the diagnosis because of the ease of biopsy in all cases of sarcoidosis of the external ear. PMID- 17125574 TI - Clinical findings and PDS mutations in 15 patients with hearing loss and dilatation of the vestibular aqueduct. AB - Following systematic skull imaging of hundred and sixty seven individuals attending a medical referral centre for the deaf in Brussels, Belgium, fifteen patients (9 per cent) aged between two and 25 years were diagnosed with dilatation of the vestibular aqueduct. Careful audiological study, with a baseline assessment then longitudinal follow up, indicated mild to profound deafness with a progressive course (i.e. an average loss of 3.3 dB per year) and frequent dizziness. Sequencing of PDS was performed in all individuals. Alterations of this gene (either homozygous, heterozygous or compound heterozygous base changes) were found in 53 per cent of patients with a large vestibular aqueduct. Four new mutations (two missense, a splice site and a four base pair insertion) were described. We were unable to confirm a correlation between homozygosity, heterozygosity and a Pendred or deafness-only phenotype. PMID- 17125575 TI - Can self assessment of communication predict hearing loss? AB - A total of 120 subjects with hearing loss (75 men, 45 women), within the age range 18-70 years (mean, 38 years), and 15 normal subjects were administered a modified Hindi adaptation of the 'self assessment of communication' hearing loss inventory. The study aimed to determine whether there was any correlation between subjects' average pure tone thresholds and their inventory scores. Data was analysed using the Pearson coefficient of correlation and regression analysis. A negative correlation was obtained stating that the greater the hearing loss, the lower the inventory score. An equation could also be derived for the bilateral symmetrical sensorineural hearing loss group and the bilateral symmetrical conductive hearing loss group to enable calculation of patients' average hearing loss from their inventory scores, in the absence of an audiogram. This could aid rehabilitation in cases with either type of hearing loss (in which no medical intervention was required) when pure tone audiometry is not possible. PMID- 17125576 TI - Evaluation of hearing loss in patients with ankylosing spondylitis. AB - AIMS: The aim of this study was to investigate hearing loss in patients with ankylosing spondylitis. STUDY DESIGN: Prospective, case-control study. METHODS: Fifty-nine ankylosing spondylitis patients (118 ears) and 52 healthy control subjects (104 ears) were included. Pure tone audiometry at 250, 500, 1000, 2000, 4000 and 6000 Hz and immittance measures, including tympanometry and acoustic reflex tests, were performed in the patients and controls. RESULTS: Sensorineural hearing loss was found in 21 patients (35.5 per cent), bilateral in 15 patients and unilateral in six. Pure tone thresholds significantly differed between patients and controls at all frequencies (p<0.05). There was no statistically significant difference between the right and the left ears' thresholds at all frequencies, except at 4000 Hz in ankylosing spondylitis patients. The right ears' thresholds were higher than those of the left ears. Patients' pure tone average (PTA) thresholds were significantly different from those of controls in all three PTA groups (i.e. 250 Hz; 500, 1000 and 2000 Hz; and 4000 and 6000 Hz) (p<0.05). The differences were most prominent in the higher frequencies. CONCLUSION: Our findings suggest a decreased hearing level in ankylosing spondylitis patients, mostly at high frequencies, although the pure tone thresholds of patients and controls significantly differed at all frequencies. PMID- 17125577 TI - Transtympanic versus intramuscular steroid administration in a histamine-induced inflammatory middle-ear model. AB - OBJECTIVES: Assessment of the histopathologic effect of transtympanic and intramuscular administration of dexamethasone in an in vivo experimental animal model of middle-ear mucosal inflammation. METHODS: Fifty healthy rabbits weighting 1500-1800 g were randomly divided in three groups. In 10 animals (control group), 0.5 ml of a 20 mg/ml histamine solution was injected transtympanically. In 20 rabbits (group A), histamine challenge followed a three day intramuscular pretreatment with dexamethasone at 1 mg/kg per day. In 20 rabbits (group B), histamine challenge followed pretreatment with dexamethasone via a transtympanic route (0.3 ml, 1.2 mg dexamethasone). Middle-ear mucosa was obtained for histopathology 30 minutes after histamine administration. The following parameters were assessed: inflammation, acute inflammatory component, presence of eosinophils, inflammatory activity and fibrosis. RESULTS: Oedema, vascular dilatation and congestion, inflammation, the presence of an acute (polymorphonuclear) inflammatory component, the presence of eosinophils, and inflammatory activity were found to be of a lesser grade in the mucosae of group B. All differences were found to be statistically highly significant (p<0.01) using the Mann-Whitney test. CONCLUSION: Our findings validate the transtympanic route of dexamethasone administration in counteracting histamine effects. PMID- 17125578 TI - Nasal insertion of St John's wort: an unusual cause of epistaxis. AB - We report a case of severe unilateral epistaxis requiring surgical arrest of bleeding, via endoscopic sphenopalatine artery ligation and anterior ethmoidal artery ligation. This followed recreational nasal insertion of St John's wort (Hypericum perforatum). Interactions between this substance and prescribed drugs have been described following oral ingestion. However, this unusual case highlights a further, worrying potential rhinological side effect of this substance. PMID- 17125579 TI - 'Complementary ENT': a systematic review of commonly used supplements. AB - OBJECTIVE: To assess the evidence surrounding the use of certain complementary supplements in otolaryngology. We specifically focussed on four commonly used supplements: spirulina, Ginkgo biloba, Vertigoheel and nutritional supplements (cod liver oil, multivitamins and pineapple enzyme). MATERIALS AND METHODS: A systematic review of the English and foreign language literature. INCLUSION CRITERIA: in vivo human studies. EXCLUSION CRITERIA: animal trials, in vitro studies and case reports. We also excluded other forms of 'alternative medicine' such as reflexology, acupuncture and other homeopathic remedies. RESULTS: Lack of common outcome measures prevented a formal meta-analysis. Three studies on the effects of spirulina in allergy, rhinitis and immunomodulation were found. One was a double-blind, placebo, randomised, controlled trial (RCT) of patients with allergic rhinitis, demonstrating positive effects in patients fed spirulina for 12 weeks. The other two studies, although non-randomised, also reported a positive role for spirulina in mucosal immunity. Regarding the use of Ginkgo biloba in tinnitus, a Cochrane review published in 2004 showed no evidence for this. The one double-blind, placebo-controlled trial that followed confirmed this finding. Regarding the use of Vertigoheel in vertigo, two double-blind RCTs and a meta-analysis were identified. The first RCT suggested that Vertigoheel was equally effective in reducing the severity, duration and frequency of vertigo compared with betahistine. The second RCT suggested that Vertigoheel was a suitable alternative to G. biloba in the treatment of atherosclerosis-related vertigo. A meta-analysis of only four clinical trials confirms that Vertigoheel was equally effective compared with betahistine, G. biloba and dimenhydrinate. Regarding multivitamins and sinusitis, two small paediatric pilot studies reported a positive response for chronic sinusitis and otitis media following a course of multivitamins and cod liver oil. Regarding bromelain (pineapple enzyme) and sinusitis, one randomised, multicentre trial including 116 children compared bromelain monotherapy to bromelain with standard therapy and standard therapy alone, for the treatment of acute sinusitis. The bromelain monotherapy group showed a faster recovery compared with the other groups. CONCLUSION: The positive effects of spirulina in allergic rhinitis and of Vertigoheel in vertigo are based on good levels of evidence, but larger trials are required. There is overwhelming evidence that G. biloba may play no role in tinnitus. There is limited evidence for the use of multivitamins in sinus symptoms, and larger randomised trials are required. PMID- 17125580 TI - Presentation of first branchial cleft anomalies: the Sheffield experience. AB - Abnormalities of the first branchial cleft are rare. They may present with a cutaneous defect in the neck, parotid region, external auditory meatus or peri auricular area, or with inflammatory or infective lesions at these sites. A retrospective case note review of the patients treated by the senior author is presented. This group consisted of 18 patients and represents the largest published UK series to date. Eleven patients (65 per cent) had undergone incomplete surgery prior to referral. Over half the patients had a clinically apparent lesion in relation to the external auditory meatus. There was a variable relationship between the tract and the facial nerve, which was identified at surgery in 15 cases. These findings are consistent with those of previously published series. Clinicians should keep this diagnosis in mind when assessing patients with infected lesions in the neck and parotid area. Surgeons should be familiar with parotid surgery, in children where appropriate, and be prepared to expose the facial nerve before embarking on the surgical management of these lesions. PMID- 17125581 TI - Role of ossiculoplasty in canal wall down tympanoplasty for middle-ear cholesteatoma: hearing results. AB - The aim of this study was to evaluate the hearing results of ossiculoplasty in canal wall down tympanoplasty in one stage middle-ear cholesteatoma surgery. We carried out a retrospective review of a consecutive series of 142 cases which had undergone type two or three canal wall down tympanoplasty with ossicular reconstruction, between January 1995 and December 2002, due to chronic otitis media with cholesteatoma.Pre-operative audiometric testing revealed a mean air conduction pure tone average (PTA) of 50.97 dB and a mean bone conduction PTA of 22.14 dB. The mean post-operative result for air conduction PTA was 37.62 and for bone conduction PTA was 23.37 dB. The mean pre- and post-operative air-bone gaps (ABGs) were 28.83 and 13.94 dB, respectively, with a gain of 14.89 dB. Almost 62.67 per cent of patients closed their ABGs to within 20 dB. Our functional results are comparable with those of other authors. In the present study, we show that hearing improvement is possible following cholesteatoma surgery with canal wall down tympanoplasty and ossicular chain reconstruction. PMID- 17125582 TI - Reconstruction of circumferential pharyngolaryngectomy using a 'horseshoe-shaped' pectoralis major myocutaneous flap. AB - Eighteen patients underwent a circumferential pharyngolaryngectomy reconstruction post pharyngolaryngectomy. All tumours showed pharyngo-oesophageal junction invasion or circular involvement of the hypopharynx, requiring total, circumferential pharyngolaryngectomy. A pectoralis major myocutaneous flap was directly sutured to the pre-vertebral fascia. A Montgomery salivary bypass tube was introduced into the oesophagus, and a nasogastric tube was placed within it. No post-operative complications occurred for 14 patients and only minor ones for the other four. Average post-operative stay was 12 days. For 15 patients, feeding returned to normal throughout the follow-up period. No fistulae were noted and neopharyngeal stenosis occurred in three patients (16 per cent). Thirteen had complete resection with normal margins. Thirteen were still alive after one to three years of follow up. The local recurrence rate was 16 per cent. The use of a pectoralis major myocutaneous flap with Spriano's technique provides a simple, reliable method for circumferential hypopharyngeal resection. It has been applied to a wide range of patients, especially elderly ones and those with poor general status, as well as to cases in which widespread involvement was detected during surgery. PMID- 17125583 TI - Mapping candidate genes for Drosophila melanogaster resistance to the parasitoid wasp Leptopilina boulardi. AB - Drosophila melanogaster resistance against the parasitoid wasp Leptopilina boulardi is under the control of a single gene (Rlb), with two alleles, the resistant one being dominant. Using strains bearing deletions, we previously demonstrated that the 55E2-E6; 55F3 region on chromosome 2R is involved in the resistance phenomenon. In this paper, we first restricted the Rlb containing region by mapping at the molecular level the breakpoints of the Df(2R)Pc66, Df(2R)P34 and Df(2R)Pc4 deficiencies, using both chromosomal in situ hybridization and Southern analyses. The resistance gene was localized in a 100 kb fragment, predicted to contain about 10 different genes. Male recombination genetic experiments were then performed, leading to identification of two possible candidates for the Rlb gene. Potential involvement of one of this genes, edl/mae, is discussed. PMID- 17125584 TI - QTL mapping of grain length in rice (Oryza sativa L.) using chromosome segment substitution lines. AB - Chromosome segment substitution (CSS) lines have the potential for use in QTL fine mapping and map-based cloning. The standard t-test used in the idealized case that each CSS line has a single segment from the donor parent is not suitable for non-idealized CSS lines carrying several substituted segments from the donor parent. In this study, we present a likelihood ratio test based on stepwise regression (RSTEP-LRT) that can be used for QTL mapping in a population consisting of non-idealized CSS lines. Stepwise regression is used to select the most important segments for the trait of interest, and the likelihood ratio test is used to calculate the LOD score of each chromosome segment. This method is statistically equivalent to the standard t-test with idealized CSS lines. To further improve the power of QTL mapping, a method is proposed to decrease multicollinearity among markers (or chromosome segments). QTL mapping with an example CSS population in rice consisting of 65 non-idealized CSS lines and 82 chromosome segments indicated that a total of 18 segments on eight of the 12 rice chromosomes harboured QTLs affecting grain length under the LOD threshold of 2.5. Three major stable QTLs were detected in all eight environments. Some minor QTLs were not detected in all environments, but they could increase or decrease the grain length constantly. These minor genes are also useful in marker-assisted gene pyramiding. PMID- 17125585 TI - Palliating the impact of fixation of a major gene on the genetic variation of artificially selected polygenes. AB - Selective sweeps of variation caused by fixation of major genes may have a dramatic impact on the genetic gain from background polygenic variation, particularly in the genome regions closely linked to the major gene. The response to selection can be restrained because of the reduced selection intensity and the reduced effective population size caused by the increase in frequency of the major gene. In the context of a selected population where fixation of a known major gene is desired, the question arises as to which is the optimal path of increase in frequency of the gene so that the selective sweep of variation resulting from its fixation is minimized. Using basic theoretical arguments we propose a frequency path that maximizes simultaneously the effective population size applicable to the selected background and the selection intensity on the polygenic variation by minimizing the average squared selection intensity on the major gene over generations up to a given fixation time. We also propose the use of mating between carriers and non-carriers of the major gene, in order to promote the effective recombination between the major gene and its linked polygenic background. Using a locus-based computer simulation assuming different degrees of linkage, we show that the path proposed is more effective than a similar path recently published, and that the combination of the selection and mating methods provides an efficient way to palliate the negative effects of a selective sweep. PMID- 17125587 TI - Neuronal-glial networks as substrate for CNS integration. AB - Astrocytes have been considered, for a long time, as the support and house keeping cells of the nervous system. Indeed, the astrocytes play very important metabolic roles in the brain, but the catalogue of nervous system functions or activities that involve directly glial participation has extended dramatically in the last decade. In addition to the further refining of the signalling capacity of the neuroglial networks and the detailed reassessment of the interactions between glia and vascular bed in the brain, one of the important salient features of the increased glioscience activity in the last few years was the morphological and functional demonstration that protoplasmic astrocytes occupy well defined spatial territories, with only limited areas of morphological overlapping, but still able to communicate with adjacent neighbours through intercellular junctions. All these features form the basis for a possible reassessment of the nature of integration of activity in the central nervous system that could raise glia to a role of central integrator. PMID- 17125586 TI - X-linked mental retardation and epigenetics. AB - The search for the genetic defects in constitutional diseases has so far been restricted to direct methods for the identification of genetic mutations in the patients' genome. Traditional methods such as karyotyping, FISH, mutation screening, positional cloning and CGH, have been complemented with newer methods including array-CGH and PCR-based approaches (MLPA, qPCR). These methods have revealed a high number of genetic or genomic aberrations that result in an altered expression or reduced functional activity of key proteins. For a significant percentage of patients with congenital disease however, the underlying cause has not been resolved strongly suggesting that yet other mechanisms could play important roles in their etiology. Alterations of the 'native' epigenetic imprint might constitute such a novel mechanism. Epigenetics, heritable changes that do not rely on the nucleotide sequence, has already been shown to play a determining role in embryonic development, X-inactivation, and cell differentiation in mammals. Recent progress in the development of techniques to study these processes on full genome scale has stimulated researchers to investigate the role of epigenetic modifications in cancer as well as in constitutional diseases. We will focus on mental impairment because of the growing evidence for the contribution of epigenetics in memory formation and cognition. Disturbance of the epigenetic profile due to direct alterations at genomic regions, or failure of the epigenetic machinery due to genetic mutations in one of its components, has been demonstrated in cognitive derangements in a number of neurological disorders now. It is therefore tempting to speculate that the cognitive deficit in a significant percentage of patients with unexplained mental retardation results from epigenetic modifications. PMID- 17125588 TI - alpha-synuclein and Parkinson's disease: the first roadblock. AB - alpha-synuclein gene mutations are major underlying genetic defects known in familial juvenile onset Parkinson's disease (PD), and alpha-synuclein is a major constituent of Lewy Bodies, the pathological hallmark of PD. The normal cellular function of alpha-synuclein has been elusive, and its exact etiological mechanism in causing dopaminergic neuronal death in PD is also not clearly understood. Very recent reports now indicate that mutant or simply over-expressed alpha- synuclein could cause damage by interfering with particular steps of neuronal membrane traffic. alpha-synuclein selectively blocks endoplamic reticulum-to-Golgi transport, thus causing ER stress. A screen in a yeast revealed that alpha- synuclein toxicity could be suppressed by over-expression of the small GTPase Ypt1/Rab1, and that over-expression of the latter rescues neuron loss in invertebrate and mammalian models of alpha-synuclein-induced neurodegeneration. alpha-synuclein may also serve a chaperone function for the proper folding of synaptic SNAREs that are important for neurotransmitter release. We discuss these recent results and the emerging pathophysiological interaction of alpha-synuclein with components of neuronal membrane traffic. PMID- 17125589 TI - Cellular secretion studied by force microscopy. AB - Using the optical microscope, real adventures in cellular research began in earnest in the latter half of the nineteenth century. With the development of the electron microscope, ultramicroscopy, and improved cell staining techniques, significant advances were made in defining intracellular structures at the nanometer level. The invention of force microscopy, the atomic force microscope (AFM) in the mid 1980s, and the photonic force microscope (PFM) in the mid 1990s, finally provided the opportunity to study live cellular structure-function at the nanometer level. Working with the AFM, dynamic cellular and subcellular events at the molecular level were captured in the mid 1990s, and a new cellular structure 'the porosome' in the plasma membrane of all secretory cells has been defined, where specific docking and fusion of secretory vesicles occur. The molecular mechanism of fusion of the secretory vesicle membrane at the base of the porosome membrane in cells, and the regulated release of intravesicular contents through the porosome opening to the extracellular space, has been determined. These seminal discoveries provide for the first time a molecular mechanism of cell secretion, and the possibility to ameliorate secretory defects in disease states. PMID- 17125590 TI - Tumor stroma fosters neovascularization by recruitment of progenitor cells into the tumor bed. AB - The tumor stroma is an active player during carcinogenesis and contains a variety of cell types such as vascular cells, fibroblasts and inflammatory cells which directly or indirectly foster neovascularization. During tumor progression stromal cells, in particular the neovasculature, acquire new characteristics distinct from their normal counterparts and display a high degree of plasticity to meet the tumor's demands. The local environment may, to some extent, shape pre existing, tumor-resident stromal cells. However, there is accumulating evidence that new endothelial and other stromal cells are actively recruited into tumors, and that this recruitment is essential for a unique and tumor-specific proangiogenic environment. PMID- 17125591 TI - Insulin-producing cells derived from stem cells: recent progress and future directions. AB - Type 1 diabetes is characterized by the selective destruction of pancreatic beta cells caused by an autoimmune attack. Type 2 diabetes is a more complex pathology which, in addition to beta-cell loss caused by apoptotic programs, includes beta cell dedifferentiation and peripheric insulin resistance. beta-Cells are responsible for insulin production, storage and secretion in accordance to the demanding concentrations of glucose and fatty acids. The absence of insulin results in death and therefore diabetic patients require daily injections of the hormone for survival. However, they cannot avoid the appearance of secondary complications affecting the peripheral nerves as well as the eyes, kidneys and cardiovascular system. These afflictions are caused by the fact that external insulin injection does not mimic the tight control that pancreatic-derived insulin secretion exerts on the body's glycemia. Restoration of damaged beta cells by transplantation from exogenous sources or by endocrine pancreas regeneration would be ideal therapeutic options. In this context, stem cells of both embryonic and adult origin (including beta-cell/islet progenitors) offer some interesting alternatives, taking into account the recent data indicating that these cells could be the building blocks from which insulin secreting cells could be generated in vitro under appropriate culture conditions. Although in many cases insulin-producing cells derived from stem cells have been shown to reverse experimentally induced diabetes in animal models, several concerns need to be solved before finding a definite medical application. These refer mainly to the obtainment of a cell population as similar as possible to pancreatic beta cells, and to the problems related with the immune compatibility and tumor formation. This review will summarize the different approaches that have been used to obtain insulin-producing cells from embryonic and adult stem cells, and the main problems that hamper the clinical applications of this technology. PMID- 17125592 TI - Adenovirus-mediated gene transfer of interleukin-4 into pancreatic stellate cells promotes interleukin-10 expression. AB - Pancreatic stellate cells (PSC) are crucially involved in the development of fibrosis, a hallmark of chronic pancreatitis. Therefore, PSC represent an attractive target for the modulation of cellular functions providing the prerequisite for the establishment of novel therapeutic strategies like transfer of genetic material to the cells. Based on recent studies suggesting that the chronic course of pancreatitis is associated with immune deviation towards a Th1 cytokine profile, we have investigated the applicability of primary PSC to an adenovirus-mediated transfer of the cDNA encoding the Th2 cytokine interleukin (IL) 4 and the autocrine-acting effects of IL 4 on the cells in vitro. The transduction of primary PSC with a replication-incompetent adenovirus type 5 vector carrying the cDNA encoding rat IL- 4 resulted in a distinct expression of the cytokine on mRNA and protein level for two weeks. Similar to recombinant IL 4, effects of the endogenously synthesized cytokine were mediated by the signal transducer and activator of transcription (STAT)6. Interestingly, beside the increase of PSC proliferation, IL 4 transduction was accompanied by an up regulation in the endogenous expression of the anti-inflammatory cytokine IL 10. In summary, our data suggest that PSC are suitable targets for gene therapy modulating cellular interactions in the pancreas. PMID- 17125593 TI - Differential proteomic profiling to study the mechanism of cardiac pharmacological preconditioning by resveratrol. AB - Recent studies demonstrated that resveratrol, a grape-derived polyphenolic phytoalexin, provides pharmacological preconditioning of the heart through a NO dependent mechanism. To further explore the molecular mechanisms involved in resveratrol-mediated cardioprotection, we monitored the effects of resveratrol treatment after ischemia-reperfusion on the protein profile by implementation of proteomic analysis. Two groups of rats were studied; one group of animals was fed resveratrol for 7 days, while the other group was given vehicle only. The rats were sacrificed for the isolated working heart preparation and for isolation of cytoplasmic fraction from left ventricle homogenates to carry out the proteomic as well as immunoblot at baseline and at the end of 30 min ischemia/2-h perfusion. The results demonstrate significant cardioprotection with resveratrol evidenced by improved ventricular recovery and reduced infarct size and cardiomyocyte apoptosis. The left ventricular cytoplasmic fractions were separated by two-dimensional electrophoresis (2-DE). Differentially regulated proteins were detected with quantitative computer analysis of the Coomassie blue stained 2-DE images and identified by MALDI-TOF (MS) and nanoLC-ESI-Q-TOF mass spectrometry (MS/MS). Five redox-regulated and preconditioning- related proteins were identified that were all upregulated by resveratrol: MAPKK, two different alphaB-crystallin species, HSP 27 and PE binding protein. Another HSP27 species and aldose reductase were downregulated and peroxiredoxin- 2 remained constant. The results of the immunoblot analysis of phosphorylated MAPKK, -HSP27 and alphaB-crystallin and PE binding protein were consistent with the proteomic findings, but not with peroxiredoxin-2. The proteomic analysis showed also downregulation of some proteins in the mitochondrial respiratory chain and matrix and the myofilament regulating protein MLC kinase-2. The results of the present study demonstrate that proteomic profiling enables the identification of resveratrol induced preconditioning-associated proteins which reflects not only changes in their expression level but also isoforms, post-translational modifications and regulating binding or activating partner proteins. PMID- 17125594 TI - Immunohistochemical evaluation of phosphorylated SMAD2/SMAD3 and the co-activator P300 in human glomerulonephritis: correlation with renal injury. AB - BACKGROUND: Smad2 and Smad3 are transcription factors that mediate transforming growth factor beta (TGF-beta) signals. Upon their activation, phosphorylated Smad2/Smad3 (pSmad2/Smad3), translocate to the nucleus and associate with co activators such as p300, regulating the transcription of genes that contribute to the fibrotic processes. METHODS: We investigated the immunohistochemical expression of pSmad2/Smad3 and the co-activator p300 in 152 renal biopsy specimens from patients with various types of glomerulonephritides (GNs) and in 15 normal kidney specimens. Patients' clinical data (serum creatinine levels and proteinuria) had been collected. RESULTS: There was a dramatic increase in the expression of pSmad2/3 and p300 in all glomerular cell types in all GNs. pSmad2/3 expression was increased in all tubular segments (except for the proximal tubules in nonproliferative GNs), while p300 expression was significantly increased only in the proximal tubular cells in all GNs. Glomerular and tubular pSmad2/Smad3 and p300 were significantly increased in proliferative GNs (compared to the nonproliferative), particularly in the secondary group. The expression profile of p300 correlated positively with the expression of pSmad2/Smad3 in the diseased glomeruli and proximal tubules. pSmad2/3 and p300 were very often detected in segmental hyperplastic lesions, cellular crescents, microadhesions and segmental or global sclerotic areas. Glomerular and proximal tubular pSmad2/Smad3 was positively correlated with serum creatinine levels, while distal and collecting tubular pSmad2/3 and p300 correlated positively with tubular atrophy. Glomerular and proximal tubular pSmad2/3 expression and glomerular p300 expression correlated positively with lupus nephritis activity. CONCLUSION: Our results suggest that pSmad2/3-p300 pathway may play a pivotal role in the pathogenesis and progression of human glomerulonephritis. PMID- 17125595 TI - Liver fibrogenesis due to cholestasis is associated with increased Smad7 expression and Smad3 signaling. AB - BACKGROUND/AIMS: Profibrogenic TGF-beta signaling in hepatic stellate cells is modulated during transdifferentiation. Strategies to abrogate TGF-beta effects provide promising antifibrotic results, however, in vivo data regarding Smad activation during fibrogenesis are scarce. METHODS: Here, liver fibrosis was assessed subsequent to bile duct ligation by determining liver enzymes in serum and collagen deposition in liver tissue. Activated hepatic stellate cells were identified by immunohistochemistry and immunoblots for alpha smooth muscle actin. Cellular localization of Smad3 and Smad7 proteins was demonstrated by immunohistochemistry. RTPCR for Smad4 and Smad7 was conducted with total RNA and Northern blot analysis for Smad7 with mRNA. Whole liver lysates were prepared to detect Smad2/3/4 and phospho- Smad2/3 by Western blotting. RESULTS: Cholestasis induces TGF-beta signaling via Smad3 in vivo, whereas Smad2 phosphorylation was only marginally increased. Smad4 expression levels were unchanged. Smad7 expression was continuously increasing with duration of cholestasis. Hepatocytes of fibrotic lesions exhibited nuclear staining Smad3. In contrast to this, Smad7 expression was localized to activated hepatic stellate cells. CONCLUSIONS: Hepatocytes of damaged liver tissue display increased TGF-beta signaling via Smad3. Further, negative feedback regulation of TGF-beta signaling by increased Smad7 expression in activated hepatic stellate cells occurs, however does not interfere with fibrogenesis. PMID- 17125596 TI - Mice with targeted disruption of spermidine/spermine N1-acetyltransferase gene maintain nearly normal tissue polyamine homeostasis but show signs of insulin resistance upon aging. AB - The N(1)-acetylation of spermidine or spermine by spermidine/spermine N(1) acetyltransferase (SSAT) is the ratecontrolling enzymatic step in the polyamine catabolism. We have now generated SSAT knockout (SSAT-KO) mice, which confirmed our earlier results with SSATdeficient embryonic stem (ES) cells showing only slightly affected polyamine homeostasis, mainly manifested as an elevated molar ratio of spermidine to spermine in most tissues indicating the indispensability of SSAT for the spermidine backconversion. Contrary to SSAT deficient ES cells, polyamine pools in SSAT-KO mice remained almost unchanged in response to N(1),N(11)-diethylnorspermine (DENSPM) treatment compared to a significant reduction of the polyamine pools in the wild-type animals and ES cells. Furthermore, SSATKO mice were more sensitive to the toxicity exerted by DENSPM in comparison with wild-type mice. The latter finding indicates that inducible SSAT plays an essential role in vivo in DENSPM treatmentevoked polyamine depletion, but a controversial role in toxicity of DENSPM. Surprisingly, liver polyamine pools were depleted similarly in wild-type and SSAT-KO mice in response to carbon tetrachloride treatment. Further characterization of SSAT knockout mice revealed insulin resistance at old age which supported the role of polyamine catabolism in glucose metabolism detected earlier with our SSAT overexpressing mice displaying enhanced basal metabolic rate, high insulin sensitivity and improved glucose tolerance. Therefore SSAT knockout mice might serve as a novel mouse model for type 2 diabetes. PMID- 17125597 TI - Role of mast cells in wound healing process after glass-fiber composite implant in rats. AB - Glass-fiber composites are frequently used in dentistry. In order to evaluate their biocompatibility we tested, in an experimental model "in vivo", their tissue response pointing our attention on presence of mast cells (MCs) and fibrotic process. Sprague Dawley rats were used for the experimental design. The fibers were introduced in a subcutaneous pocket along the middle dorsal line between the two scapulas for 7, 14 or 21 days. At the end of the treatments the skins were excised and then processed for Toluidine Blue, to determine the presence of MCs, and Picrosirius Red staining, to evaluate the presence of fibrotic tissue. Our preliminary results showed and increase of both MC number and deposition of collagen type I, which characterized the fibrotic tissue. So, subsequent aims of our study were to evaluate the role played by MCs in tissue fibrosis and to give a possible explanation regarding the mechanisms that were responsible of biological response observed, through the analyses of some proteins, such as metalloproteinase-2 (MMP-2), its inhibitor (TIMP-2) and transforming growth factor-beta (TGF-beta). Our data confirmed the involvement of TGF-beta, released by MCs, in the disruption of the equilibrium between MMP-2 and TIMP-2 that were implicated in the enhancement of fibrosis. In summary, this study demonstrate that this type of materials induced an inflammatory response at the site of implant and help to clarify what type of mechanism and which proteins are involved in this biological response. Nevertheless, more extensive investigations are in progress to better evaluate the inflammatory process. PMID- 17125598 TI - The IL-6 promoter polymorphism is associated with disease activity and disability in systemic sclerosis. AB - Systemic sclerosis (SSc) is a rare, autoimmune disease characterized by cutaneous and visceral fibrosis. Interleukin- 6 (IL-6) is involved in the pathogenesis of many immune-mediated diseases. IL-6 plays an important role in the initiation and promotion of fibrosis. The polymorphism in the position -174 (G/C) of the promoter region of the IL-6 gene (IL-6pr) may alter the expression of the gene. Complete linkage disequilibrium was observed between the -174 and -597 alleles. The aim of this study is to investigate the possible influence of -597 (-174) IL 6pr polymorphism on the susceptibility and/or the clinical course of SSc in Romanian population. Genotyping of -597 variant was performed by an RFLP method on 20 SSc patients and 26 healthy subjects. Patients having the homozygous GG ( 597) genotype had higher disease activity and disability scores than heterozygous GA patients: the European Scleroderma Study Group (EScSG) disease activity score was 5.0 +/- 3.3 in homozygous GG subjects vs. 2.4 +/- 3.6 in heterozygous GA patients (p < 0.05), and the Disability Index of the Health Assessment Questionnaire (HAQ-DI) was 1.42 +/- 1.04 in homozygous GG subjects vs. 0.53 +/- 0.55 in heterozygous GA patients (p < 0.05). No difference was observed in the distribution of allele frequencies between SSc patients and healthy controls. CONCLUSIONS: The GG homozygosis was found to be associated with a higher degree of illness activity and disability in SSc patients. No statistically significant differences were found between SSc patients and healthy controls with respect to the -597 allele distribution. PMID- 17125599 TI - Caveolae in smooth muscles: nanocontacts. AB - Smooth muscle cell (SMC) caveolae have been investigated by quantitative and qualitative analysis of transmission electron microscopy (TEM) images of rat stomach, bladder and myometrium, guinea pig taenia coli, human ileum, and rat aortic SMCs. Ultrathin (below 30 nm) serial sections were used for examination of caveolar morphology and their connections with SMC organelles. Average caveolar diameter was smaller in vascular SMCs (70 nm, n=50) than in visceral SMCs (77 nm, n=100), but with the same morphology. Most of the caveolae, featured as flask shaped plasma membrane (PM) invaginations, opened to the extracellular space through a 20 nm stoma (21 +/- 3 nm) having a 7 nm thick diaphragm. A small percentage of caveolae (3%), gathered as grape-like clusters, did not open directly to the extracellular space, but to irregular PM pockets having a 20-30 nm opening to the extracellular space. In visceral SMCs, caveolae were disposed in 4-6 rows, parallel to myofilaments, whilst aortic SMCs caveolae were arranged as clusters. This caveolar organization in rows or clusters minimizes the occupied volume, providing more space for the contractile compartment. The morphometric analysis of relative volumes (% of cell volume) showed that caveolae were more conspicuous in visceral than in vascular SMCs (myometrium - 2.40%; bladder - 3.66%, stomach - 2.61%, aorta - 1.43%). We also observed a higher number of caveolae per length unit of cellular membrane in most visceral SMCs compared to vascular SMCs (myometrium - 1.06/microm, bladder - 0.74/microm, aorta - 0.57/microm, stomach - 0.48/microm). Caveolae increase the cellular perimeter up to 15% and enlarge the surface area of the plasma membrane about 80% in SMCs. Threedimensional reconstructions (15micro(3)) showed that most caveolae, in both visceral and vascular SMCs, have nanocontacts with SR (87%), other with mitochondria (10%) and 3% apparently have no contact with these organelles. Usually, 15 nm wide junctional spaces exist between caveolae and SR, some of them with nanostructural links between each other or with mitochondria: direct contacts (space <2 nm or none) and molecular links, so called 'feet' (about 12 nm electron dense structures between organellar membranes). Direct contacts possibly allow molecular translocation between the two membranes. Electron-dense 'feet' like structures suggest a molecular link between these organelles responsible for intracellular Ca(2+) homeostasis (excitation-contraction coupling or pharmaco mechanical coupling). Close appositions (approximately 15 nm) have also been observed between caveolae and perinuclear SR cisternae, suggesting that caveolae might be directly implicated in excitation-transcription coupling. PMID- 17125600 TI - Cutaneous melanoma: digital dermoscopy-essential tool for positive diagnosis. AB - Cutaneous melanoma is a "perfid", aggressive and hard to be treated malignant tumor in case of delayed diagnosis. However, patients still have a chance to escape progressive disease if the lesion is recognized early, when the surgical approach is curative. Dermoscopy has the important advantage of rapidity and non invasivity in a field with (still) contradictory algorithms of diagnosis and treatment. The recognition of the elementary dermoscopic lesions enables accurate diagnosis for cutaneous melanoma. In our opinion, dermoscopy appears compulsory in the routine dermatologic examination. In vivo microscopy (dermoscopy) together with histopathology (plus or minus immunohistochemistry) seem, at present, to provide the most reliable diagnosis of melanoma. PMID- 17125602 TI - [Strategies for treatment of chronic hepatitis B virus infection]. PMID- 17125601 TI - Interstitial cells of Cajal (ICC) and gastrointestinal stromal tumor (GIST): facts, speculations, and myths. AB - Interstitial cells of Cajal (ICC) is a peculiar cell network composed of cells having processes described by the eminent Spanish neuroanatomist of the 19th century, S. Ramon y Cajal. ICC became a fascinating subject to many investigators and it is estimated that there are over 100 publications yearly on the subject related to ICC, in the last three years. Now it is widely accepted that ICC are pace maker cells of the gut and probable progenitor cells of gastrointestinal stromal tumors (GIST). Lately, interstitial Cajal-like cells (ICLC) are being found in various organs and their physiological role is still to be defined. We have reviewed the literature trying to evaluate the validity of the current concept and found that there are a few salient points to be considered. 1) There has been some important departure in defining the identity of ICC from the original criteria of Cajal. In particular, ICC with myoid feafures in intestinal smooth muscle layers (ICC-DPM) do not seem to fit to the original description of interstitial cell network by Cajal. We have also pointed out that the current reports assigning a pace maker role to ICC vastly depend on the scientific data on "ICC with myoid features", not on "fibroblast-like ICC", which are more abundant and easier to identify. 2) There seem to be an overwhelming amount of data proving the relationship between ICC and GIST. Both are known to express c Kit and the ultrastructural characteristics seen in GIST roughly parallel those of ICC including minimal myoid differentiation seen in the majority of GIST, supporting the current concept that GIST are ICC tumors. 3) According to the original description of Cajal, ICC was not limited to the gut, suggesting an existence of ICC in other organs. The list of organs reported to contain ICC (currently identified by immunohistochemistry and electron microscopy) is ever growing and further studies are needed to define their identity and pathophysiologic role. 4). Recent data concerning gut development suggest that both c-Kit expressing ICC (fibroblasts-like as well as muscle-like) and gut muscle cells derive from the common progenitor cells of the embryonic gut unifying the histogenetic concept of all GIST with heterogeneous cytomorphologic features. In this review we attempted to incorporate recent information on interstitial Cajal-like cells (ICLC) found in other organs to broaden our understanding of ICC in general in terms of their ultrastructure, physiology, and neoplasia. PMID- 17125603 TI - [A double-blind, randomized, lamivudine-controlled clinical trial of DAIDING (adefovir dipivoxil) for lamivudine-resistant patients with chronic hepatitis B]. AB - OBJECTIVE: To investigate the efficacy and safety of adefovir dipivoxil (ADV, DAIDING) for Chinese chronic hepatitis B patients with lamivudine (LAM) resistance. METHODS: This study was a multicenter, double-blind clinical trial. 209 chronic hepatitis B patients with LAM resistance were randomly put in an ADV, DAIDING or a LAM group. After 24 and 48-weeks of treatment, serum HBV DNA levels were measured by quantitative PCR and liver function tests; HBV serology and safety assessments were also conducted. RESULTS: The mean reduction of HBV DNA from baseline at 24 and 48 weeks was significantly greater in the ADV group compared with that in the LAM group (2.40 log10 vs 0.94 log10, P < 0.01; 2.71 log10 vs 1.07 log10, P < 0.01). In the ADV group, the virological response and ALT normalization at 24 and 48 weeks were significantly higher than those in the LAM group. There was no significant difference between the two groups in the portion of HBeAg reduction, HBeAg seroconversion and incidence of adverse events. There was no severe adverse event related to the investigational product, DAIDING, in this trial. CONCLUSION: DAIDING (ADV) is effective and safe for the treatment of chronic hepatitis B patients with LAM resistance. PMID- 17125604 TI - [Pegylated interferon alpha 2a in treating chronic hepatitis B patients]. AB - OBJECTIVE: To evaluate the efficacy and safety of pegylated interferon alpha 2a (PEG-IFN alpha-2a) in treating patients with chronic hepatitis B. METHOD: Seventy two patients with chronic hepatitis B were assigned to a PEG-IFN alpha-2a (experimental) group (n=42) and an interferon alpha (control) group (n=30) randomly. Each patient in the experimental group received 180 microg PEG-IFN alpha-2a every week. Each patient in the control group received 500 MU interferon alpha every day. All the patients were treated for 48 weeks, and then were followed for another 48 weeks with no treatment. RESULTS: At the end of the 12th week, the rate of HBeAg negative cases was 30% in the PEG-IFN alpha-2a group, which was much higher than in the control group (x2 = 4.162, P < 0.05). The values of HBeAg and the log value of HBV DNA in the PEG-IFN alpha-2a group were much lower than the values before the treatment (t = 2.689, t = 4.080, P <0.01), but there was no difference between before and after treatment in the control group ( t = 1.229, t = 1.009, P > 0.05). At the end of the 24th week, the rate of HBeAg negative cases in the PEG-IFN alpha-2a group was much higher than that in the control group (x2=6.190, P < 0.05). The value of HBeAg and the log value of HBV DNA in the PEG-IFN alpha-2a group were much lower than in the control group (t=2.215, t=2.122, P < 0.05). At the end of the 48th week, besides the reduction mentioned above, the rate of cases with HBeAg/antiHBe seroconversion and normalization of ALT and complete responsiveness in the PEG-IFN alpha-2a group were all much higher than those in the control group (x2=5.771, x2=5.617, x2=5.308, P < 0.05). At the end of 48 weeks with no treatment, all the parameters mentioned above in the PEG-IFN alpha-2a group were much better than those in the control group and they remained so, but they were different in the control group (x2=11.943, t=3.439, t=6.111, x2=9.930, x2=9.522, x2=7.920, P < 0.01). Nine patients in the PEG-IFN alpha-2a group had liver biopsies before their treatment and also at the end of their treatment. The expressions of HBsAg and HBcAg were decreased at the end of the treatment. The rate of expression of HBsAg in the liver tissues before the treatment was 88.9% but only 22.2% at the end of the treatment (x2=8.001, P < 0.01). The rate of expression of HBcAg in the livers before treatment was 66.7% but only 33.3% at the end of the treatment. Before and at the end of the PEG-IFN alpha-2a treatment, there were no significant changes in the degrees of inflammation and fibrosis and the quantity of collagen in the liver tissues. Three patients in the PEG-IFN alpha-2a group (10%) were HbsAg negative. Two of them were found so at the end of 32 weeks with treatment and one patient was found at the end of 24 weeks with no treatment, but there were no HBsAg negative patients in the control group. The adverse reactions that occurred in the PEG-IFN alpha-2a and in the control groups were similar. CONCLUSION: PEG IFN alpha-2a was effective in inhibiting HBV replication. The effect of PEG-IFN alpha-2a was lasting. PEG-IFN alpha-2a was well tolerated during our treatment. PMID- 17125605 TI - [Study on the pharmacokinetic profile of telbivudine]. AB - OBJECTIVE: To evaluate the pharmacokinetic profile of telbivudine in healthy Chinese subjects after oral administration of single and multiple doses. METHODS: Forty-two healthy adult male and female subjects 18-40 years of age were randomized into four telbivudine dosing groups of 200 mg, 400 mg, 600 mg and 800 mg. Subjects in the 600 mg group received both a single dose and once daily multiple doses for 8 consecutive days. Telbivudine concentrations in plasma and urine samples collected at different time points before and after the drug administration were measured using HPLC-MS/MS. Pharmacokinetic parameters were calculated by using the non-compartmental approach. RESULTS: After a single dose of 200 mg, 400 mg, 600 mg and 800 mg, tmax (median) were 2.50, 2.00, 2.00 and 2.50 hours respectively; t1/2 were (43.3 +/- 15.2) h, (49.1 +/- 14.4) h, (39.4 +/ 12.1) h and (46.7 +/- 20.8) h respectively; Cmax were (1,753.2 +/- 389.0) ng/ml, (2,586.7 +/- 871.4) ng/ml, (3,703.6 +/- 1,219.0) ng/ml and (3454.6 +/- 953.9) ng/ml respectively; AUC(0-infinity) were (12,843.2 +/- 2,925.6) ng.h(-1).ml(-1), (22,948.9 +/- 5,721.0) ng.h(-1)/ml(-1), (26,440.5 +/- 8,938.1) ng.h(-1).ml(-1) and (28, 820.9 +/- 7 912.9) ng.h(-1).ml(-1) respectively, and CL(R) (600 mg) was (6,545.6 +/- 1 504.4) ml/h. The AUCss from multiple doses was (1,088.5 +/- 299.8) ng/ml; Cmax and AUC accumulation ratio were 1.02 +/- 0.21 and 1.23 +/- 0.26 respectively, which implicated moderated accumulation. CONCLUSION: Pharmacokinetic parameters of telbivudine in Chinese healthy subjects were determined. PMID- 17125606 TI - [Foscarnet sodium for treatment in patients with severe chronic hepatitis B]. AB - OBJECTIVE: To investigate the effectiveness of foscarnet sodium in the treatment of severe chronic hepatitis B. METHODS: Two hundred and eight patients were enrolled in a multicenter, double-blind, controlled study. The patients received foscarnet sodium (foscarnet group) or saline (control group) injections for 4 weeks, and were then followed for 24 weeks. RESULTS: HBV DNA negative rate was 12.8% in the foscarnet group and 7.1% in the control group at the end of treatment; and it was 5.5% and 3.0% at the end of the follow-up period respectively (P > 0.05). The rate of HBV DNA decrease of more than 2 log copies/ml was 53.2% in the foscarnet group and 16.2% in the control group at the end of treatment, and 23.9% and 8.1% (P < 0.01) respectively at the end of the follow-up period. The rate of HBV DNA < 10(5) copies/ml was 64.2% and 30.3% at week 4 in the two groups respectively, and 40.4% and 22.2% (P < 0.01) at the end of the follow-up period. HBeAg negative rate was 17.3% and 5.8% at the end of the treatment, and 22% and 5.4% at the end of the follow-up period (P < 0.01). The rate of HBeAg seroconversion was 12.7% and 3.7% at week 4, and 16.7% and 1.5% at the end of the follow-up period. Response rate was 60.6% and 21.2% at the end of week 4 (P < 0.05). CONCLUSION: Foscarnet sodium injection has a good effect on severe chronic hepatitis B patients and it is safe to use on them. PMID- 17125607 TI - [Expression of endotoxin receptors on hepatic stellate cells during their activation]. AB - OBJECTIVE: To investigate the role of endotoxin receptor expression in the activation of hepatic stellate cells (HSCs). METHODS: HSCs were isolated from normal rats and the expression of endotoxin receptors on quiet HSCs and in vitro activated HSCs was determined using RT-PCR and immunocytochemical staining methods. A rat model of liver fibrosis and cirrhosis was established. The expressions of CD14 and alpha-SMA in liver tissues were detected by immunohistochemical staining. RESULTS: Freshly isolated HSCs had a low level of CD14 mRNA expression and no expression of TLR4 mRNA was detected. The in vitro activated HSCs had increased expressions of CD14 mRNA and TLR4 mRNA and LPS up regulated the expression of endotoxin receptors. Immunocytochemical staining showed cytoplasmic and nucleolus staining for CD14 in the cultured HSCs. LPS played a further role on CD14 protein expression. In the development of liver fibrosis, the number of CD14-positive cells in the livers was increased and these cells were distributed along the sinusoids. In the later stage of liver fibrosis, the CD14-positive cells were gathered in the fibrotic septae, which also contained alpha-SMA positive cells. CONCLUSION: The activated HSCs expressed endotoxin receptors. The endotoxin receptors may be involved in the role in which HSCs played in the inflammatory process and liver fibrosis development. PMID- 17125608 TI - [Fasudil inhibits HSC adhesion, migration and proliferation via Rho/ROCK pathway]. AB - OBJECTIVES: To observe the effects of fasudil, a Rho/ROCK signaling pathways inhibitor, on adhesion, migration and proliferation of hepatic stellate cells (HSCs). METHODS: Cultured HSCs were divided into 5 groups. Fasudil was added to 4 groups in a concentration of 12.5, 25, 50, and 100 micromol/L, respectively. A group without fasudil added served as untreated controls. The adhesive inhibition effect of fasudil on HSCs was examined by toluidine blue colorimetric assay, the inhibition of migration of HSCs was evaluated by a modified Boyden chamber, and cell proliferation was assessed by MTT assay. The protein levels of RhoA, p-MLC (Thr18/Ser19) and alpha-SMA were assayed by Western blot. RESULTS: Fasudil inhibited HSC adhesion, proliferation and LPA-induced migration in a concentration-dependent manner; the protein expressions of alpha-SMA and p-MLC (Thr18/Ser19) were significantly decreased in the presence of fasudil. CONCLUSION: Fasudil can inhibit HSC adhesion, migration and proliferation by suppressing the cytoskeleton regulation function of Rho/ROCK signaling pathways. PMID- 17125609 TI - [Exploration of differential expressed genes involved in the development and progression of hepatocellular carcinoma using oligo microarray]. AB - OBJECTIVES: To analyze the differential expression genes (DEGs) among hepatocellular carcinoma (HCC), para-cancerous tissue (PCT) and normal liver tissue (NLT) and explore the target genes related to the development and progression of HCC. METHODS: The total RNAs of matched HCC, PCT and NLT of HCC patients were isolated using one step Trizol method. Matched RNAs were qualified using 10 g/L agarose gel electrophoresis and lab-on-chip. cRNAs were synthesized, fluorescence labeled and purified after total RNAs were purified. The RNAs of HCC and NLT, HCC and PCT were hybridized with Agilent oligo microarray (21,074 probes). The fluorescence intensity features were detected by Agilent scanner and quantified by feature extraction software. The selected candidate genes were confirmed by SYBR Green I stained real time RT-PCR. RESULTS: (1) The total RNA, reverse transcription product and fluorescence labeled cRNA were all of high quality; (2) There were 420 up-regulated genes and 552 down-regulated genes among 2-fold DEGs, including DKK1 (dickkopf homolog 1) which was 5-fold up-regulated; (3) The results of real time RT-PCR, using beta-actin as an internal control, showed that the 2-Delta Ct values of DKK1 in HCC, PCT and NLT were 0.089 504, 0.007,65 and 0.000,631 respectively. CONCLUSION: (1) The high throughput and effective Agilent oligo microarray can screen novel therapy targeted genes by analyzing the DEGs in development and progression of HCC; (2) The development and progression of HCC is a complicated process involving multigenes and multiprocedures; (3) DKK1, as a novel gene, is involved in the development and progression of HCC and may be a new therapy target. PMID- 17125610 TI - [Relationship between leptin gene of adipose tissues and nonalcoholic fatty liver disease]. AB - OBJECTIVE: To investigate leptin mRNA expressions in subcutaneous (SC) and omental (OM) adipose tissues of patients with nonalcoholic fatty liver disease (NAFLD), and their relationships with insulin resistance (IR), blood leptin, blood triglyceride, total blood cholesterol, blood glucose, body weight index and waist-hip ratio. METHODS: SC and OM adipose tissues were obtained from 10 obese and 11 nonobese NAFLD patients and from 11 obese and 13 nonobese patients without NAFLD, who served as controls. Leptin mRNA expression levels in the subcutaneous and omental adipose tissues were measured using SYBR Green I quantitative real time PCR. IR was estimated using homeostasis assessment (HOMA). The levels of plasma leptin and insulin were measured using ELISA. RESULTS: The relative mRNA expression of leptin, HOMA-IR and blood leptin levels in NAFLD differed significantly from those of the controls (P < 0.05). The leptin/GAPDH ratio of the obese and nonobese NAFLD and control cases were 1.32 +/- 0.12, 0.99 +/- 0.05, 1.10 +/- 0.09, 0.87 +/- 0.13 respectively. The expression levels of SC and OM adipose leptin mRNA in NAFLD patients were positively correlated with HOMA-IR (r=0.72, P < 0.05), blood leptin (r=0.69, P < 0.05), blood triglyceride (r=0.32, P < 0.05), body weight index (r=0.57, P < 0.05) and waist-hip ratio (r=0.50, P <0.05). CONCLUSION: The primary reason for high levels of blood leptin is high leptin mRNA expression in adipose tissues; in both obese and nonobese patients with NAFLD; high levels of blood leptin and the leptin mRNA expression in adipose tissues and IR exist. These findings suggest that leptin resistance exists in patients with NAFLD and leptin resistance is positively correlated with NAFLD, the same as in insulin resistance. PMID- 17125611 TI - [An analysis of 276 cases of drug-induced liver damage]. AB - OBJECTIVE: Drug-induced liver damage is a potential complication from using many drugs. The aim of our study was to analyze the etiology and clinical features of drug-induced liver damage, in order to draw more attention to this problem. METHODS: Two hundred and seventy-six cases over a 5-year period in Jiangsu Province Hospital were retrospectively analyzed. RESULTS: A variety of drugs, including traditional Chinese medicines (26.1% of our total cases) and anticancer drugs (17%) caused liver damage. The main clinical manifestations of it were fatigue, nausea, vomiting and jaundice. In 88% of our cases the symptoms were relieved or completely disappeared, but there was still a 5.1% mortality rate. CONCLUSIONS: The clinical features of drug-induced liver damage are of no specificity, and the mortality of it is not low. Liver function should be monitored when suspected drugs are prescribed. PMID- 17125612 TI - [How long should patients with chronic hepatitis B virus infection be treated]. PMID- 17125613 TI - [Focusing on the markers for monitoring and antiviral therapy in chronic hepatitis B virus infection]. PMID- 17125614 TI - [Antiviral treatment for hepatitis B virus related liver cirrhosis]. PMID- 17125615 TI - [The dynamic changes of hepatitis C virus quasispecies during natural infections]. PMID- 17125616 TI - [The effects of transforming growth factor beta1/Smads signaling pathway changes on the pathogenesis of liver fibrosis]. PMID- 17125617 TI - [Expression of angiotensinogen during development and spontaneous reversal of hepatic fibrosis]. PMID- 17125618 TI - [Expression of melittin tagged with green fluorescent protein and its use in hepatocellular carcinoma treatment]. PMID- 17125619 TI - [The expression of CYP2E1 and its effect on the proliferation of C3A hepatocyte cell line]. PMID- 17125620 TI - [Effect of NAC on LPS-induced toll-like receptor 2 and 4 expressions in murine macrophages]. PMID- 17125621 TI - [Therapeutic effects of Trigonella foenum-greacum saponin on rats with alcoholic fatty livers]. PMID- 17125622 TI - [Screening of target autoantigen in hepatocytes from hepatocyte cDNA library by phage display technique]. PMID- 17125623 TI - [The distribution and significance of insulin-like growth factor binding protein in patients with liver fibrosis]. PMID- 17125624 TI - [Correlation between plasma soluble thrombomodulin level to the degree of liver inflammation in patients with chronic viral hepatitis]. PMID- 17125625 TI - [The effects of refecoxib on inhibiting the growth and inducing apoptosis of hepatocellular carcinoma cell lines HepG2 and human normal liver cell line QSG7701]. PMID- 17125626 TI - [An introduction of a treatment algorithm for the management of chronic hepatitis B virus infection in the USA]. PMID- 17125627 TI - [Progress in human hepatic progenitor cell research]. PMID- 17125628 TI - [Recent advances in research of molecular mechanism in alcoholic fatty liver disease]. PMID- 17125629 TI - Primary care research: opportunities and challenges. PMID- 17125630 TI - Private drug sellers' education in improving prescribing practices. AB - OBJECTIVE: To determine the education of private drug sellers as an intervention tool in promoting rational use of medicines for diarrhoea at private drug outlets in a rural setting. PLACE AND DURATION OF STUDY: The study was conducted at Mithi, Tharparkar in rural Sindh in August 2005. PATIENTS AND METHODS: Diarrhea was selected to study drug-prescribing habit of private drug sellers. Evidence based guidelines for control of diarrhea were developed along with simulation cases and keys. In phase-1, simulators were sent to 6 private drug sellers and their prescription was noted. After phase-1, 3 private drug sellers were educated about prescribing, while in the other 3, there was no intervention. In phase-2, simulators were sent again and prescriptions by private drug sellers was noted. RESULTS: The work experience of private drug sellers was between 5 to 15 years. Two private drug sellers were employed at each of the drug outlet with education between matriculation and graduation. All the private drug sellers prescribed medicines to customers with confidence. The private drug sellers prescribed medicines to customers on a daily basis that included all categories of medications. Amoebicidals, Anti-diarrheals, antispasmodics, antibiotics, analgesics and oral rehydration solutions were prescribed by 22(38.6%), 10(17.5%), 3(4.9%), 10(17.5%), 3(5.3%) and 9(15.8%) private drug sellers prior to intervention respectively. It changed to 0(0%), 6 (14%), 0(0%) 0 (0%), 4 (9.3%) and 16(37.2%) postintervention respectively. Referral to hospital for serious cases increased after intervention. CONCLUSION: The rational use of medicines can be promoted by providing education to private drug sellers. Further studies and education programs for private drug sellers are strongly recommended across the country. PMID- 17125631 TI - Fixed drug eruption: topical provocation and subsequent phenomena. AB - OBJECTIVE: To determine the usefulness of topical provocation in detecting the incriminated drug causing fixed eruption. DESIGN: Quasi-experimental study. PLACE AND DURATION OF STUDY: Department of Dermatology, Shaikh Zayed FPGMI, Lahore, from November 2002 to December 2005. PATIENTS AND METHODS: Three hundred and five, clinically diagnosed cases of Fixed Drug Eruption (FDE) of either gender and of any age were subjected to topical provocation with different drugs by using concentration of 1% (n=203), 2% (n=210) and 5% (n=235) in white soft paraffin. Drug ointment of one strength was applied one at a time on normal skin of flexor surface of right or left forearm. The effects of tests on involved and uninvolved skin were observed for 48 hours. The changes in lesions like erythema, hyperpigmentation, itching, burning or appearance of new lesion were considered a positive response. In case of no change, the patients (n=5) were subjected to oral provocation test, by giving half to full therapeutic dose of the suspected drug depending upon the severity of the initial attack. A patient who exhibited see-sawing phenomenon with 5% metamizole TPT was given oral challenge with same drug. Control topical tests were repeated in equal number of normal persons with various drug ointments and in patients of FDE with white soft paraffin on normal and affected skin. RESULTS: One hundred and thirty-seven patients were males and one hundred and sixty-eight patients were females. Maximum number of patients belonged to third decade. With 1% drug preparations 12 out of 316, with 2% drug preparations 28 out of 422 and with 5% drug preparations, 312 out of 523 TPTs were positive. The comparison revealed a highly significant association (Chi square 448.1 and p < 0.000) among various strengths of preparations and positive response. Sulphamethoxazole was found to be the most commonly incriminated cause of FDE applied in 5% concentration yielded sensitivity rate of 91% compared to 4% with lower concentrations. Positive patch test was also observed with oxytetracycline. Five patients who were given oral provocation with different drugs were found to be positive to tinidazole, dapsone, propylphenazone, belladonna and phenobarbitone. Interesting phenomena like earlier reactivation of lesion situated distal to site of application of preparation, marching, see sawing (with oral metamizole) and lightening observed, are not yet reported in literature. CONCLUSION: Topical provocation test with 5% drug concentration applied on normal skin of patient with FDE is a possible first line investigation in finding the incriminated drug. PMID- 17125632 TI - Pattern of thyroid autoimmunity in type 1 and type 2 diabetics. AB - OBJECTIVE: To determine the level of thyroid autoimmunity among clinically euthyroid patients of type 1 and type 2 diabetics and to correlate the levels with pattern of diabetes. DESIGN: Cross-sectional analytical study. PLACE AND DURATION OF STUDY: Diabetic Clinic of Shaikh Zayed Hospital, Lahore from August 2004 to April 2005. PATIENTS AND METHODS: Blood samples were collected from 163 subjects (51 of type 1, 61 of type 2 diabetics and 51 non- diabetic controls) and sera were separated. They were all clinically euthyroid and selected conveniently. Thyroid profile (FT4, TSH and anti-TPO) was assessed in the three groups and compared. RESULTS: It was observed that at a cut off value of less than 100 units of anti-TPO level was normal, raised anti-TPO antibodies were found in 12% of normal control group, 61% in type 1 and 42.3% in type 2 diabetics. In type 1 diabetics, the TSH was higher in the younger age patients with progressive rise in anti-TPO antibodies level by age. The TSH, anti-TPO antibodies and BSF were significantly higher in type 1 diabetics than in the controls. In type 2 diabetics, TSH was moderately low but anti-TPO antibodies and BSF were higher than in the controls. CONCLUSION: Thyroid autoimmune process seems to be correlated more with type 1 diabetic patients than with type 2 patients. PMID- 17125633 TI - Hemodynamic effects of terlipressin in patients with bleeding esophageal varices secondary to cirrhosis of liver. AB - OBJECTIVE: To study the hemodynamics of terlipressin in bleeding esophageal varices due to cirrhosis of the liver. DESIGN: A quasi-experimental study. PLACE AND DURATION OF STUDY: Medical-ICU, Jinnah Postgraduate Medical Centre, Karachi from February to July 2005. PATIENTS AND METHODS: Seventy-eight consecutive patients with bleeding esophageal varices were evaluated. The diagnosis of cirrhosis was based on history, physical examination, laboratory data and abdominal ultrasound. Blood-pressure and pulse rate were monitored. Injection terlipressin 2 mg intravenous bolus was given followed by 2 mg i/v 6 hourly. Intravenous plasma expanders, whole blood, fresh frozen plasma and platelet concentrates were transfused as needed. Upper gastrointestinal endoscopy was performed for evaluation and grading of varices, detection of portal gastropathy, and banding. Descriptive and inferential statistics were applied as applicable. RESULTS: Seventy patients of either gender, aged 18 - 95 years were included in the study. Systolic blood-pressure(SP) increased by 7.77 mmHg (mean SP: 108.1 mmHg, SD + 9.84, 95% CI: 105.77 - 110.43 mmHg; p-value: 0.0002); diastolic blood pressure(DP) by 21.57 mmHg (mean DP: 79.71 mmHg, SD + 7.35, 95% CI: 77.97 - 81.45 mmHg; p-value: 0.001) and mean arterial pressure by 9.42 mmHg(mean MAP: 89.12 mmHg, SD + 6.98, 95% CI: 87.45 - 90.78 mmHg; p-value: 0.0007) within 24 hours of initiating terlipressin in majority of patients. The pulse rate decreased in 34 (48.5%) patients by 6-24 beats/min in 30 min, and by 2-12 beats/min in 24 hours; and increased in 30 (42.85)% patients by 10-15 beats/min at 30 min and by 2-8 beats/min at 24 hours. CONCLUSION: Terlipressin had an overall benefit in significant number of patients with bleeding esophageal varices in this series and only a few adverse effects when given at recommended doses. PMID- 17125634 TI - Role of hysterosalpingoscintigraphy in the workup of infertility. AB - OBJECTIVE: To assess the role of hysterosalpingoscintigraphy (HSSG) in the evaluation of fallopian tube patency and function and compare the results with hysterosalpingography (HSG) and laparoscopy (LS). DESIGN: Cross-sectional study. PLACE AND DURATION OF STUDY: The study was conducted at Multan Institute of Nuclear Medicine and Radiotherapy (MINAR), Multan from August 2004 to February 2005. PATIENTS AND METHODS: HSSG was performed after instillation of 4mCi (148 MBq) 99mTechnetium-macroaggregated albumin (99mTc-MAA) in posterior vaginal fornix in 65 patients. Serial static images were acquired in supine position at 1 hour, 2 hours, 3 hours and, if needed, at 24 hours. The results were compared to the findings on LS and HSG. RESULTS: Out of 65 patients, 37 (56.9%) patients had bilateral blocked tubes, 17 (26.1%) patients had bilateral patent tubes, 6 (9.2%) patients had blocked left tube and 5 (7.1%) patients had blocked right tube. The calculated sensitivity, specificity, positive predicted value (PPV), negative predicted value (NPV) and accuracy for HSSG were 90%, 83%, 90% and 90% respectively. The agreement between HSSG and LS was found in 32 out of 35 patients and agreement between HSG and HSSG was found in 24 out of 30 patients. CONCLUSION: This simple procedure can delineate tubal physiology; in selected cases it can replace HSG and in others augment the information gathered by HSG. HSSG should be part of the infertility workup algorithm. PMID- 17125635 TI - Management of intermittent angle closure glaucoma with Nd: YAG laser iridotomy as a primary procedure. AB - OBJECTIVE: To assess the efficacy and complications of Nd: YAG laser iridotomy in patients with intermittent (sub-acute) angle closure glaucoma. STUDY DESIGN: An interventional study. PLACE AND DURATION OF STUDY: The Ophthalmology Unit-1, Civil Hospital and Dow Medical College, Karachi, from February 2000 to February 2002. PATIENTS AND METHODS: Twenty-five eyes of twenty-three patients with periodic (intermittent) angle closure, selected in outpatient department, were kept on pilocarpine until YAG laser iridotomy was performed. After YAG laser iridotomy oral acetazolamide and topical dexamethasone was used to control postlaser rise of IOP and inflammation respectively. Patency of iridotomy was confirmed and intra-ocular pressure was measured one hour after the procedure. Immediate complication, if any, was noted. Follow-up was done for six months. Prophylactic laser iridotomy was done in fellow eye with occludable angle. Levene's test for equality of variance and t-test for equality of means were used for statistical analysis. RESULTS: This study revealed a significant difference in IOP before and after YAG laser iridotomy (p = .002). Complete follow-up of 6 months was possible in 25 eyes of 23 subjects. After YAG Laser iridotomy, 21 (84%) eyes showed negative provocative test, intra-ocular pressure below 19 mmHg without medication and anterior chamber angle no more occludable and were labeled successful. Iridotomy remained patent in 96% of eyes. Iridotomy failed to reduce IOP in 4 (16%) eyes. The complications were minimal and transient. CONCLUSION: YAG laser iridotomy offers effective, long lasting, first line treatment for the management of primary angle closure glaucoma at intermittent stage. Laser iridotomy widens drainage angle and reduces IOP, once synechial angle closure occurs in more than one quadrant. PMID- 17125636 TI - Comparison of clinical causes of death with autopsy diagnosis using discrepency classification. AB - OBJECTIVE: To determine the usefulness of autopsy findings in the quality improvement of patients care. DESIGN: An observational study. PLACE AND DURATION OF STUDY: Departments of Pathology and Medicine, Combined Military Hospital (CMH) Kharian, a tertiary care hospital, from January 2001 to December 2003. MATERIALS AND METHODS: The clinical and necropsy findings of all the cases, who died in hospital and had undergone autopsy examination at CMH, Kharian, from January 2001 to December 2003, were retrieved from record of clinical case sheet data and autopsy record of the hospital. The two were analyzed and compared according to the discrepancy classification. The exclusion and inclusion criteria, the international classification of disease (ICD) to code deaths, the global burden of disease (GBD) system to classify and group diseases, and the Goldman discrepancy classification to compare clinical and autopsy diagnosis and classify the discrepancies, were used as described. RESULTS: The death rate varied from 0.94% to 1.29% and autopsy rate from 4.69% to 10.10% annually between January 2001 and December 2003. The number of cases classified according to GBD system was 3 (5%) in Group 1, 26 (43.33 %) in Group 2 and 31 (51.66 %) in Group 3. The discrepancy classes included 9 (15 %) class I major discrepancies and 3 (5 %) class II major discrepancies. Non-discrepant diagnosis was seen in 37 cases (61.66 %) and 11 cases (18.32 %) were non-classifiable. CONCLUSION: This study showed the usefulness of autopsy findings in the quality improvement of the diagnosis and management of the disease by showing only a minority of cases with discrepant diagnosis of the cause of death. PMID- 17125637 TI - Outcome of tissue sparing surgical intervention in mine blast limb injuries. AB - OBJECTIVE: To describe the pattern of mine blast limb injuries in civilian population of Kashmir, to evaluate the outcome of tissue sparing surgical intervention in these injuries and to determine the sensitivity of hand-held percutaneous Doppler for tissue viability. DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Combined Military Hospital, Muzaffarabad and Islamic International Medical College Hospitals at Rawalpindi/Islamabad from November 1997 to May 2005. PATIENTS AND METHODS: One hundred and three patients who sustained mine blast injuries to upper or lower limbs, along side the line of control between the Indian-held Kashmir and Azad Kashmir, regardless of age and gender, were included in this study. Patients who already had amputation after injury at some other place were excluded. All patients were initially managed in emergency and had more than one surgical intervention. Transcutaneous Doppler was used to evaluate the vascularity of the remaining tissue. All patients were operated under spinal or general anaesthesia and had repeated debridements followed by skin cover by split skin graft, full thickness skin graft or rotational flaps. Every patient received at least 5 days course of antibiotics and tetanus prophylaxis. Postoperative rehabilitation and follow-up was conducted for at least 6 months after discharge from the hospital. RESULTS: Mean age of victims in this study was 22 years. Out of 103 patients, 72 (69.9%) received initial wound care in the peripheral primary health care centre but were not amputated while 31 patients (30%) were just dressed and referred for further treatment at tertiary care hospitals. Eighty-five patients (82.5%), out of the total, had some sort of traumatic amputation at presentation due to the original injury. That included loss of limb below knee in 19 (18.45%) patients, at distal tibiofibular region in 13 (12.6%), mid tarsal amputations in 39(37.9%), and hemi foot amputation in 15 (14.6%) patients. Nine (8.7%) patients had losses of two or less than two toes, 1 (0.97%) patient had injury at mid palmer region, and 5 (4.9%) patients had 2 fingers traumatic amputation. Eighteen (17.5%) patients had soft tissue ( with or without bony injury) injury only without any actual traumatic amputation. Infection rate was 27% in patients who did not have wound care in the periphery. Those who had wound toilet in-the peripheral hospital had infection rate of 16%. Two patients developed fulminating multi-resistant progressive infection requiring below knee amputation. CONCLUSION: Conservative wound debridement and early skin coverage by different means and preserving maximum soft tissues and bone results in functionally and cosmetically better limb in victims of land mine. PMID- 17125638 TI - Pulmonary edema and lung injury after severe laryngospasm. AB - A young male with no pre-operative medical illness underwent corrective surgery for a deviated nasal septum under general anesthesia. At the end of surgery, patient was extubated but went into severe laryngospasm that did not improve with gentle Intermittent Positive Pressure Ventilation (IPPV) and small dose of Suxamethonium. As the situation worsened and patient developed severe bradycardia and de-saturation, re-intubation was done that revealed pink froth in the endotracheal tube. His portable chest X-ray was suggestive of non-cardiogenic pulmonary edema. With an overnight supportive treatment, using mechanical ventilation with Positive End-Expiratory Pressure (PEEP), morphine infusion and frusemide, patient improved and was subsequently weaned off from ventilator. PMID- 17125639 TI - Alpha(2)-blocker helps to avoid systemic to pulmonary shunt in a prostaglandin dependent infant with critical pulmonary valve stenosis. AB - A 27 days old newborn with critical pulmonary valve stenosis remained prostaglandin (PGE(1)) dependent for 2 weeks after successful balloon valvuloplasty. Only the introduction of Phentolamine in his medication regimen, allowed PGE(1) to be weaned off within days of this therapy. The medication was continued for 4 days and replaced by angiotensin converting enzyme inhibitor (Captopril). Few weeks after the discharge, the patient remained clinically stable with acceptable saturation. PMID- 17125640 TI - Dentigerous cyst presenting as facial pain. AB - A rare case is presented in which a maxillary dentigerous cyst had eroded the posterior wall of the right maxillary sinus into the pterygo-palatine fossa causing facial pain due to pressure on the nerves. It had also eroded the lateral wall of sinus and into the oral cavity and got infected resulting in foul smelling oral discharge. The case was dealt with complete removal of cyst using Caldwell Luc's approach. PMID- 17125641 TI - Enzyme replacement therapy in a child with Gaucher disease. AB - This case report describes an experience of using recombinant enzyme in a child who was diagnosed as a case of Gaucher disease at the age of 3 years. Regular enzyme replacement therapy has resulted in marked improvement in his hemoglobin level, absolute neutrophil count, platelets and physical growth. PMID- 17125642 TI - An unusual presentation of testicular tumor. AB - A case of testicular choriocarcinoma is reported in which blood mixed stools and haemoptysis were the presenting manifestations as the patient never told about the testicular swelling to his parents. Orchidectomy was performed but the patient presented again with massive hematemesis due to gastric perforation secondary to gastric metastasis. The size of the testis at diagnosis was approximately 12 x 7 cm. This was also unusual as testicular choriocarcinoma presents as a small mass. The patient eventually died of the complications within one month of diagnosis. PMID- 17125643 TI - Knowledge, attitude and practices of reproductive age females about tetanus toxoid vaccine: a pilot study. PMID- 17125644 TI - Oral stereognostic ability: a test of oral perception. AB - Oral stereognosis (form or shape recognition in the oral cavity) can be utilized to assess oral perceptual abilities. This assessment can be beneficial in planning as well as predicting the future outcome of any treatment modality in the orodental region. This article provides a meta analysis of different studies conducted in the last fifty years to quantify magnitude of oral sensory capacity using oral stereognostic ability tests and their implication in clinical dental sciences. PMID- 17125645 TI - Malignant melanoma in accessory nipple in a male. PMID- 17125646 TI - Not 'surviving sepsis' in the developing countries. PMID- 17125648 TI - [Something for something--physicians, drug industry and general welfare]. PMID- 17125649 TI - [How participation in a clinical trial affects doctors' choice of treatment]. AB - In a retrospective cohort study, the effects of participation in a company sponsored clinical trial on asthma drug prescribing were investigated. Ten trial conducting practices were compared with 165 control practices. Conducting the trial did not influence guideline adherence measured as the prevalence of inhaled corticosteroid use among users of inhaled beta2-agonists. However, participation in the trial led to an increased preference for sponsor's drugs. PMID- 17125650 TI - [Conflict of interest information--more stringent rules in the Ugeskrift]. PMID- 17125651 TI - [Ivabradine--a medical alternative for patients with chronic stable angina pectoris]. AB - Procoralan (Ivabradine) is a novel, specific If--current inhibitor, inducing reduction in heart-rate through a direct effect on sinoatrial automaticity at rest and during exercise. Thereby the drug may prevent myocardial ischaemia. So far the indications for using Procoralan are the following: symptomatic treatment of chronic stable angina pectoris in patients with sinus rhythm, in whom betablockers are contraindicated or not tolerated. PMID- 17125652 TI - [Clinical leaders' and staff's assessment of the current quality related to the patient pathways and expectations of and knowledge about accreditation--a baseline evaluation in Copenhagen County]. AB - INTRODUCTION: In the preparation for accreditation in Copenhagen County the commitment of clinical leaders and staff members is crucial. The objectives of these surveys are to examine the leaders' and the staff's assessment of quality improvement and their expectations and knowledge about accreditation, as well as the staff's advice concerning the further planning. MATERIALS AND METHODS: Two surveys among clinical leaders and staff members were carried out. RESULTS: Statistically, significant differences between staff and leaders were found in many areas concerning quality improvement and knowledge about accreditation. Leaders and staff both had high expectations of the use of accreditation as a tool for quality improvement, thus no statistically significant difference between expectations were found. CONCLUSION: The overall positive expectation for accreditation as a tool for quality improvement is an excellent basis for the accreditation process. The different assessments in quality among leaders/staff and positions show the need to involve all personnel in the organization. A survey about patient experiences includes the same subjects as the survey among staff and leaders. A striking discrepancy between the evaluation of quality by patients and by the leaders and staff in specific areas was found; therefore further investigations will be carried out. PMID- 17125653 TI - [Fatal pulmonary thromoembolism]. AB - INTRODUCTION: Fatal pulmonary thromboembolism is a relatively common autopsy finding. The aim of this investigation is to describe the magnitude of the problem in an autopsy material and factors of prophylactic importance and to discuss the importance of the autopsy. MATERIAL: Deceased with lethal pulmonary thromboembolism diagnosed by forensic autopsy at the Department of Forensic Medicine at the University of Southern Denmark in 1992-2005, in total 58 individuals. Data were collected from autopsy- plus police reports and medical records. METHOD: Descriptive case-only investigation. RESULTS: The pulmonary embolism had in none of the cases been diagnosed before death. Two of the individuals in this investigation were healthy young females who used oral contraceptives, and one was a psychiatric patient who died from pulmonary embolism after five days of physical restraint. CONCLUSION: The diagnosis is sometimes not made before the autopsy. The decreasing autopsy frequency may result in a lack of diagnosis of pulmonary embolism as cause of death. PMID- 17125654 TI - [Duration of treatment in private psychiatric practices]. AB - INTRODUCTION: The purpose of the study was to investigate duration of treatment and dropout from treatment in private psychiatric practices. MATERIALS AND METHODS: Data from the data base: Quality Assurance in Danish Psychiatric Private Practice (QAD3P) is applied for the investigation. The database was established in 1996 and includes information about approximately 40,000 treatment episodes. The present investigation uses information about 6,067 treatment episodes from 10 private practising psychiatrists, all of whom participated continuously in the Quality Assurance data base in the period 1998-2002 inclusive. RESULTS: Depression, anxiety and personal disorders constitute 82% of the treated episodes. Schizophrenia spectrum disorders show the longest duration of treatment (33.4% are treated for longer than 12 months), and persons with personality disorders have the shortest treatment duration (66.9% terminate the treatment within 6 months). Dropout from treatment is predicted (logistic regression) by gender (male), OR 1.31, 95% CI, 1.15-1.48 and age (< or = 44 years) (OR 2.49, 95% CI 2.21-2.81). CONCLUSION: Approximately 25% of the patients drop out of treatment. The causes for this are not known, but are correlated to patients' young age (< or = 44 years) and gender (male). PMID- 17125655 TI - [Serrated, hyperplastic and hyperplasia-like colorectal polyps]. AB - The colorectal hyperplastic polyp has generally been considered a finding of no clinical significance. Recent research has, however, called attention to the existence of some variants of hyperplastic polyp which are potentially malignant. The term "advanced serrated polyp" has been coined for such cases, which comprise mixed hyperplastic/adenomatous tissue, serrated adenoma, and sessile serrated polyp, in contrast to the traditional hyperplastic polyp. Since epithelial dysplasia is an integrated component of mixed hyperplastic/adenomatous polyp and of the serrated adenoma, such a diagnosis would dictate control colonoscopy comparable to the guidelines for subjects with conventional adenomas. The cytology of the sessile serrated polyp is, however, closer to that of the traditional hyperplastic polyp, whereas the architecture mimics that of the serrated adenoma. For this reason, a consensus regarding the optimal management of such patients has not been obtained, but if the polyp is sizeable and located in the right colon, control should be considered. The small, usually left-sided traditional polyp as a rule needs no follow-up, but the context in which such a lesion is found and its morphology may influence the clinical decision. Future large-scale investigations of serrated colorectal polyps, including interobserver studies, will be required to identify histological details of clinical utility which can be adopted in daily routine practice. PMID- 17125656 TI - [Hyperplastic colorectal polyposis]. AB - Two cases of hyperplastic polyposis are presented, a 59-year-old female with >50 colorectal serrated polyps in a pancolic distribution and a 69-year-old female with four mucinous colorectal carcinomas and seven serrated polyps, three of which >10 mm. The pathological tissue of the latter patient was confined to the right colon and the neoplastic areas displayed loss of MLH1 expression. Three first degree relatives of this patient had carcinoma of the large bowel, breast and ovary, respectively. A first degree relative of the former patient succumbed to a colorectal carcinoma. PMID- 17125657 TI - [Development of thrombi on an Amplatzer PFO Occluder]. AB - In a 37-year-old female with previous paradoxical embolism and an identified PFO, a transcatheter closure of the defect was performed with an Amplatzer PFO Occluder. The patient suffered additional embolic episodes, and three months after the implantation, echocardiography showed several thrombi attached to the right side of the occluder, despite continuous administration of aspirin and the absence of significant coagulation disorders. The PFO may have been an innocent bystander, and the thrombogenic potential of the device and the optimal antithrombotic treatment need further investigation. PMID- 17125658 TI - [Addison's disease and infection]. AB - Addison's disease, or primary adrenocortical insufficiency, is a diagnosis that may often be overlooked. A case is presented of a 28-year-old male who, within a six-month period, was admitted on three occasions with symptoms mimicking sepsis, for which no microbiological etiology could be established. Each time he recovered, however, on antibiotic treatment. Not until the third admission was Addison's disease suspected. PMID- 17125659 TI - [Fructose 1,6-bisphosphatase deficiency as a cause of recessive serious hypoglycaemia]. AB - Fructose 1,6-bisphosphatase (FBPase) deficiency is an autosomal recessive disorder of gluconeogenesis. Here we describe a family from Morocco with parental consanguinity with three affected children. All were homozygous for a novel mutation in exon 5: 685 C-->T of the gene coding for the liver isoform of fructose 1,6-bisphosphatase (FBP1). The mutation changed the amino acid codon (Q229X) from a glutamine (CAG) in position 229 to a stop codon (TAG), which caused a shortening of the protein from the normal 338 amino acids to 228. The shortened protein lacks a major part of the active site and is therefore probably without enzymatic activity. PMID- 17125660 TI - [Primary cytomegalovirus infection and thrombophlebitis/pulmonary embolism]. AB - A young woman was admitted to hospital with suspected pyelonephritis. Due to prolonged fever, further investigations were done and showed a thrombosis in her femoral vein; X-ray and lung scintigraphy revealed a pulmonary embolism. Blood tests showed lymphocytosis, and a primary cytamegalovirus (CMV) infection was confirmed by serology and PCR. The patient was a smoker and obese and was taking oral contraceptives; a factor V Leiden mutation was also found. Deep-vein thrombosis is a rare but severe complication of a primary CMV infection that is also seen in immunocompetent persons. PMID- 17125661 TI - [Quality assessment program of the Spanish Society of Infectious Diseases and Clinical Microbiology. Analysis of results. 2005]. AB - Quality assurance of the analytical processes performed at the clinical microbiology laboratory is mandatory and should be carried out by using external and internal quality control activities. External quality assessment programs allow intercomparison within laboratories, detection of errors, and evaluation of the suitability of some reagents or diagnostic kits for the purpose for which they were designed; these activities are also useful for continuous education. The program launched 15 years ago by the Spanish Society of Infectious Diseases and Clinical Microbiology is based on sending typified materials along with a clinical and microbiological case related to these control materials. The spectrum of the samples is broad, including bacteriology (monthly and three monthly), serology, mycology, parasitology, mycobacteria, virology, and molecular microbiology. After receiving the results from the participants, the program organization delivers an individual certificate comparing the results with those of a reference laboratory. Additionally, a report is generated by analyzing all the results sent by the participants; laboratories are also sent review articles on the subject of each assessment as a tool for continuous education in clinical microbiology. In this article, the most relevant conclusions and lessons from the 2005 assessments are presented. PMID- 17125662 TI - [Proteus penneri]. AB - Proteus penneri, formerly P. vulgaris biogroup 1, was recognized as a new species in 1982. This species is associated with clinical processes similar to those involving P. mirabilis and P. vulgaris and expresses similar pathogenic determinants. In clinical samples, P. penneri is mainly isolated from urine (50%), wound and soft tissue exudates (25%), and blood cultures (15%), mostly of nosocomial origin. Although P. penneri is easy to identify, it can be misidentified as P. vulgaris by automatic systems that do not include the indol test result in the identification process. This species has a characteristic susceptibility profile, essentially due to the production of the chromosomal inducible beta-lactamase HugA, which presents a high homology (86%) with CumA from P. vulgaris. HugA is inhibited by clavulanic acid and determines resistance to aminopenicillins and first- and second-generation cephalosporins, including cefuroxime, but does not affect cephamycins or carbapenems, and is inhibited by clavulanic acid. HugA is derepressed due to mutational processes in gene regulators, affecting the activity of cefotaxime and, to a much lesser extent, that of ceftazidime and aztreonam. This phenotype resembles the production of an extended spectrum beta-lactamase. Like other Proteus species, P. penneri is resistant to tetracyclines and should be considered resistant to nitrofurantoin. PMID- 17125663 TI - [Current situation of the epidemiology of meningococcal disease]. AB - Neisseria meningitidis, the etiological agent of all forms of meningococcal disease, is still a cause for concern among society in general and especially among health workers. Several of its antigens have been used in strain characterization, and some have served as the basis for the development of vaccines. In this sense, the best known are the capsular polysaccharide, which defines the serogroups, the outer membrane protein of class 2/3, used for serotype classification, and the class 1 porins, defining the serosubtype. During the last 30 years, most cases of meningococcal disease in Spain have been due to serogroup B strains, with the exception of 1996 and 1997, when serogroup C cases became the most frequent. The capsular polysaccharide has been successfully used in the development of conjugate vaccines highly effective against A, Y and W135 serogroups and particularly against serogroup C isolates. The development of a vaccine against serogroup B strains for routine immunization is still uncertain. However, the use of specific vaccines based on antigenic formulations of class 1 protein, to be applied in epidemic situations, is closer to becoming a reality. Because of the current absence of a universal vaccine against N. meningitidis, specific surveillance programs are required to evaluate the importance of recombination processes affecting capsular expression. This type of event could produce new strain variants able to avoid the immunological response generated after vaccination. PMID- 17125664 TI - [Serologic diagnosis of Mycoplasma pneumoniae infections]. AB - Mycoplasma pneumoniae is a human pathogen with worldwide distribution. This microorganism is a common cause (10-30%) of community-acquired pneumonia, also called primary atypical pneumonia because of the spectrum of clinical and radiological findings. The immune response is mainly based on rapid antibody production against peptide and glycolipid antigens derived from this microorganism. During the primary infection, IgM levels generally rise within the first week, and are then followed by an IgG response. Titers of IgG and IgA increase in reinfections. Microbiological diagnosis is based on specific antibody detection. Polymerase chain reaction (PCR) techniques performed on sputum or pharyngeal/nasopharyngeal exudates, as well as the development of multiplex PCR reactions allowing identification of M. pneumoniae and other respiratory pathogens, would by highly useful in routine diagnosis. The most common serological techniques are complement fixation, immunofluorescence, particle agglutination, and enzyme immunoassay. Diagnosis should be performed by selecting the most appropriate test according to functional criteria and population groups. Specific detection of IgM antibodies should not be included in the differential diagnosis in adults and young people. Diagnostic criteria including seroconversion or rising IgG titers may not be clinically useful, because of the time delay and the difficulty of obtaining a second serum specimen for testing, given the mildness of the clinical symptoms. PMID- 17125665 TI - [Erythrovirus B19 infection]. AB - Erythrovirus B19 has been associated with an expanding range of clinical disorders since its identification as the etiological agent of erythema infectiosum, or fifth disease of childhood: acute arthropathy, dermatologic manifestations, chronic anemia in immunocompromised patients, and transient aplastic crisis in individuals with underlying chronic hemolytic disorders. Furthermore, exposure to and infection by B19 virus can lead to serious complications during pregnancy, which may result in fetal anemia, spontaneous abortion, and hydrops fetalis. Consequently, the B19 immune status of pregnant women should be routinely determined. Because many immunocompromised patients with chronic anemia will respond positively to intravenous immunoglobulin therapy, laboratory confirmation of B19 infection is required. Since Erythrovirus B19 cannot be routinely grown in vitro, diagnostic methods for detecting the presence of B19 by molecular techniques or by investigating the specific immune response should be considered in clinical microbiology laboratories. PMID- 17125666 TI - [Diagnostic, clinical and therapeutic implications of viral hepatitides in HIV infected patients]. AB - Viral hepatitis B (HBV) and C (HCV) are a significant cause of morbidity and mortality in patients coinfected with human immunodeficiency virus (HIV). In HBV/HIV coinfection, there is a higher frequency of HBV replication, and higher rates of HBV-associated liver disease. The only drugs currently approved for the treatment of HBV infection are lamivudine, adefovir, entecavir, and interferon-a. HIV/HBV coinfection is associated with an increased frequency of hepatotoxicity in patients receiving highly active antiretroviral therapy (HAART), and reactivation of clinical hepatitis is observed upon stopping HBV-active anti-HIV drugs. Liver disease due to HCV infection is currently the leading cause of mortality among HIV-infected patients in the developed world. The treatment of choice of chronic hepatitis C in these patients is based on pegylated interferon in combination with ribavirin, which achieves sustained virological response rates of up to 40%. However, patients with HCV/HIV coinfection show accelerated progression to cirrhosis and are at increased risk of hepatotoxicity from HAART. PMID- 17125667 TI - [Changing epidemiological aspects of candidemia and their clinical and therapeutic implications]. AB - Candida species are a major cause of healthcare-related bloodstream and invasive infections. Studies assessing nosocomial bloodstream infections during the two last decades ranked Candida species as the fourth most common nosocomial bloodstream pathogen. The incidence of Candida species has risen steadily during this period due to the increase in the number and type of patients at risk for these yeasts. Infections caused by Candida are especially frequent and serious in onco-hematological patients. Over the past decade, the introduction of azole antifungals as prophylactic agents, together with other factors, has led to a shift in the species of Candida that cause infection. During the period under review (1996 to 2005) several studies have confirmed the impact of antifungal prophylaxis with azoles on the emergence of Candida species other than Candida albicans. The widespread use of fluconazole has contributed to a relative decrease in the prevalence of C. albicans, while species inherently less susceptible, such as Candida glabrata and Candida krusei, appear to be isolated with greater frequency. Moreover, laboratory studies to determine the antifungal susceptibilities and virulence of non-albicans Candida species have enabled the design of microbe-specific management strategies. More of these studies will be necessary as we enter an age in which multiple antifungal compounds (echinocandins, new azoles) will become available for clinical use in invasive candidiasis or candidemia. The present review aims to highlight the different trends in the incidence, distribution and behavior of Candida bloodstream infections in the distinct types of patients at risk. PMID- 17125668 TI - [Keratitis due to Acanthamoeba]. AB - Free-living amebae appertaining to the genus Acanthamoeba, Naegleria and Balamuthia are the most prevalent protozoa found in the environment. These amebae have a cosmopolitan distribution in soil, air and water, providing multiple opportunities for contacts with humans and animals, although they only occasionally cause disease. Acanthamoeba spp. are the causative agent of granulomatous amebic encephalitis, a rare and often fatal disease of the central nervous system, and amebic keratitis, a painful disease of the eyes. Keratitis usually follows a chronic course due to the delay in diagnosis and subsequent treatment. The clear increase in Acanthamoeba keratitis in the last 20 years is related to the use and deficient maintenance of contact lenses, and to swimming while wearing them. The expected incidence is one case per 30,000 contact lens wearers per year, with 88% of cases occurring in persons wearing hydrogel lenses. This review presents information on the morphology, life-cycle and epidemiology of Acanthamoeba, as well as on diagnostic procedures (culture), appropriate antimicrobial therapy, and prevention measures. PMID- 17125669 TI - [New methods for mycobacteria identification]. AB - Currently, the genus Mycobacterium comprises more than 100 different species, many of which cause significant clinical infections with high morbidity and mortality. Mycobacteria identification by conventional methods (rate and optimal temperature of growth, pigment production, colony morphology, and biochemical characteristics) has been the standard in most clinical microbiology laboratories. However, this phenotypic approach has considerable limitations, since numerous species cannot be differentiated. Moreover, because of the slow growth of these microorganisms, the results may not be available until 2-4 weeks after the initial isolation. Therefore, one of the most important challenges for clinical mycobacteriology laboratories is rapid and accurate identification of this variety of microorganism. This review aims to briefly describe several alternative procedures for mycobacterial identification. Although analysis of cell wall lipids (mycolic acids) by high-performance liquid chromatography is an interesting and well-known option, the most promising innovation for mycobacteria identification is the use of rapid molecular methods such as nucleic acid probes and, especially, genomic amplification methods. PMID- 17125670 TI - [Diagnostic utility of nucleic acid detection by real-time polymerase chain reaction]. PMID- 17125671 TI - [CMV DNA detection in plasma using real-time PCR based on the SYBR-Green I dye method]. AB - OBJECTIVE: The aim of this study is to assess a real-time PCR technique on the LightCycler 2.0 with SYBR-Green I detection as compared to another real-time PCR method based on detection with FRET (fluorescence resonance energy transfer) probes for the quantification of CMV DNA. METHODS: The two real-time PCR methods were used to test plasma samples from immunocompromised patients with clinically suspected CMV disease, patients under follow-up without symptoms, and healthy adults. A standard curve for quantitative analysis by the SYBR-Green I method was performed with 10-fold diluted solutions of DNA from the CMV Towne strain (ATCC VR-977) cultured in MRC-5 monolayer. In addition, frozen samples from patients positive for CMV pp65 antigenemia were also analyzed and results compared using the two real time PCR methods. RESULTS: The real-time PCR technique using SYBR Green I on the LightCycler 2.0 was a highly specific, fast, simple and reliable test to quantify CMV; moreover, it was cost-effective. CONCLUSION: Quantification of CMV DNA in plasma using this sensitive, fast, low-cost method was advantageous for the diagnosis and follow up of patients with opportunistic CMV infection, which are increasingly more frequent in our daily hospital clinical practice. PMID- 17125672 TI - [Fosfomycin trometamol: multiple-dose regimen for the treatment of lower urinary tract infections]. AB - INTRODUCTION: A short antibiotic regimen is recommended for the treatment of uncomplicated lower urinary tract infection. Nevertheless, the treatment to follow in other situations is not so clearly defined. When the person affected by lower urinary tract infection is not a young woman, it is recommended to treat at least 7 days, and quinolones or cotrimoxazole are the antibiotics most often used. However, because of the frequency of drug resistance in this type if infection, it is advisable to apply antibiotics with lower rates of resistance, such as fosfomycin trometamol, for longer treatment periods than the often-used single dose. METHODS: Using the data on urinary elimination of fosfomycin after a single dose obtained in a prior study in healthy volunteers, we simulated the urinary concentrations of this antibiotic following administration of two doses. In addition, we calculated the interval of administration required to achieve urinary concentrations greater than 16 mg/L, the critical concentration of sensitivity for Escherichia coli, one of the most commonly implicated microorganisms in these infections. RESULTS: Fosfomycin concentrations in urine persisted above the defined cut-off for 161 hours after administration of two 3-g doses of fosfomycin trometamol, 72 hours apart. This implied an efficacy time of 66% in a period of 7 days. CONCLUSION: From the pharmacokinetic viewpoint, the optimum dosage of fosfomycin trometamol to achieve appropriate urinary concentrations along 7 days is administration of two 3-g doses, 72 hours apart. PMID- 17125673 TI - [Assessment of the invasiveness of rapidly-growing nonpigmented mycobacteria with the fibroblast microcolony assay]. AB - OBJECTIVE: The invasive capacity of rapidly-growing nonpigmented mycobacteria strains was evaluated by means of a fibroblast microcolony assay and related to the colony phenotype on Middlebrook 7H11 and to the clinical significance of the isolates. METHODS: Twenty-nine strains [Mycobacterium chelonae (8), M. fortuitum (6), M. peregrinum (5), M. abscessus (5), M. mucogenicum (4) and M. septicum (1)], proceeding from a bacterial collection and clinical isolates, were evaluated. The smooth or rough phenotype of the colonies was assessed in Middlebrook 7H11 medium. Intracellular invasiveness was determined by the fibroblast-microcolony assay described by Shepard. Quantitative culture characteristics were compared with Student's t-test, and qualitative characteristics with Fisher's exact test. RESULTS: No significant differences were found between colonies with different phenotypes or strains having a different clinical significance, except for two strains of Mycobacterium chelonae isolated from cases of catheter-related bacteremia, which showed elongated microcolonies. M. fortuitum and M. peregrinum strains showed larger microcolonies than M. chelonae, M. abscessus and M. mucogenicum, and displayed a fluffy appearance, while the latter two strains showed rounded colonies. CONCLUSION: Very few strains of mycobacteria had invasive capacity and the majority of strains isolated from human infections do not show this characteristic; hence this trait is not essential for mycobacteria to cause infection in humans. PMID- 17125674 TI - [Etiology of acute viral respiratory tract infections in children from Gran Canaria, the Canary Islands (Spain)]. AB - OBJECTIVE: Acute respiratory tract infections (ARTI) of viral origin are a frequent cause of pediatric consultations and hospital admissions. The aim of this study was to investigate the etiology of these infections in Gran Canaria, the Canary Islands, (Spain). METHODS: From May 2002 through May 2005, 1957 nasopharyngeal washings were collected from 1729 children presenting with ARTI to the Pediatric Emergency Unit. A rapid antigen detection method was performed in every sample to identify respiratory syncytial virus (RSV). An immunofluorescence assay (IFA) and cell culture (CC) was used in RSV-negative samples. RESULTS: Median age was 2 months (range, 0.03-119). A viral agent was identified in 1032 children (59.7%). RSV was detected in 769 children (74.5%). Other viruses identified, in order of frequency, were parainfluenza viruses, rhinoviruses, adenoviruses, influenza viruses, enteroviruses, and coronaviruses. Statistical differences were found between age and the type of virus detected: Adenoviruses caused respiratory infections in older children (median age: 6 months; range: 1 74). There were 6 mixed infections. Sensitivity of IFA as compared to CC was 55.8%, and specificity was 99.2%. CONCLUSIONS: Respiratory viruses are responsible for a large number of ARTI cases in children from Gran Canaria, RSV being the major cause. Viral identification is determinant for managing these patients and making a proper use of antibacterial and antiviral drugs. PMID- 17125675 TI - [Streptococcus agalactiae highly resistant to fluoroquinolones]. AB - BACKGROUND: Streptococcus agalactiae remains susceptible to penicillin; nevertheless, an increase in the resistance to other antimicrobial families, such as macrolides and more rarely fluoroquinolones, has been described. METHODS AND RESULTS: From 2003 to 2004, two fluoroquinolone-resistant S. agalactiae strains were detected. The strains presented one mutation in parC (Ser79 --> Phe) and an additional mutation in gyrA (Glu85 --> Ala or Glu85 --> Lys). CONCLUSION: This study describes the first S. agalactiae strains highly resistant to fluoroquinolones in Spain. PMID- 17125676 TI - [Assessment of two methods of antigenic detection by ELISA for the diagnosis of norovirus outbreaks]. AB - INTRODUCTION: The aim of this study was to assess two ELISA techniques for the detection of outbreaks of norovirus. METHOD: One-hundred and sixty-five fecal samples from patients affected in 30 outbreaks were studied. RESULTS: On the basis of a specific consensus criterion between techniques for confirming outbreaks, the sensitivity and specificity was respectively 80% and 90% for RIDASCREEN, and 80% and 100% for IDEIA. CONCLUSION: These techniques could be useful for rapid detection of norovirus outbreaks in laboratories that lack molecular confirmation techniques. PMID- 17125677 TI - [Expert recommendations for the diagnosis and treatment of chronic hepatitis C infection in the prison setting]. AB - The prevalence of HCV infection in Spanish prisons is very high (38.5%). The characteristics of the infected patients, particularly the high rate of HIV coinfection, makes it very likely that the morbidity and mortality produced by serious liver disease secondary to this infection will increase considerably in the coming years. A group of Spanish experts with experience in patients who are inmates has been invited to establish a series of recommendations for the diagnosis and treatment of chronic hepatitis C infection in Spanish prisons. PMID- 17125678 TI - [Hepatitis B and C virus antiviral resistance]. AB - Infection by the hepatitis B (HBV) and C (HCV) viruses is a major cause of morbidity and mortality world-wide. The clinical outcomes of infection by these viruses (e.g., chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma) depend on several factors related to the host and the viral agent. Among the latter, factors associated with the response to current antiviral therapies, such as the emergence of resistance mutants and the genotype responsible for the infection, are gaining increasing importance. As has been established for human immunodeficiency virus (HIV), the presence of resistance mutations in the viral polymerase constitutes the main problem for treating HBV infection with approved drugs and those recently applied. Methods have been developed to detect these mutations, as well as algorithms to predict the response to treatment. The outcome of treatment for HCV infection is highly influenced by the viral genotype, however, and our understanding of the molecular basis for the response to interferon in these patients has grown considerably in recent years. PMID- 17125680 TI - [Advanced HIV infection and diffuse hyperkeratosic skin lesions in a male patient]. PMID- 17125679 TI - [Hepatic abscess due to Klebsiella pneumoniae in an Asian immigrant]. PMID- 17125681 TI - [Middle lobe bronchiectasis and Mycobacterium avium complex infection: the Lady Windermere syndrome]. PMID- 17125682 TI - [Lymphadenopathies caused by Toxoplasma gondii in a general hospital]. PMID- 17125685 TI - [Prosthetic joint infection: possible role of perioperative allogeneic blood transfusion]. PMID- 17125683 TI - [Broad-spectrum beta-lactamases in enterobacteria from hospitalized and ambulatory patients in Zaragoza (Spain), 2001-2004]. PMID- 17125684 TI - Cerebral phaeiohyphomycosis due to Cladophialophora bantiana. PMID- 17125686 TI - [Cryptococcal disease and immunosuppressive treatment]. PMID- 17125687 TI - Old pathogens and old drugs. PMID- 17125689 TI - [Detecting asthma in schools]. PMID- 17125688 TI - [Endocarditis due to Bartonella spp. Three new clinical cases and Spanish literature review]. PMID- 17125693 TI - [Obstructive sleep apnea-hypopnea syndrome in children is not associated with obesity]. AB - OBJECTIVE: The prevalence of obstructive sleep apnea-hypopnea syndrome (OSAHS) in the general pediatric population ranges from 1% to 3%. However, its prevalence in an unselected population of obese children is unknown. We studied the association between obesity and OSAHS in children diagnosed with the syndrome in a cohort of boys and girls (age range, 2-14 years) referred to the pediatric respiratory medicine outpatient clinic at our hospital for suspected apnea, snoring, or both over the past 5 years. PATIENTS AND METHODS: The medical history of each patient was recorded and all patients underwent a physical examination, chest and nasal cavities radiography, and 8-channel respiratory polygraphy during sleep. The following variables were evaluated: sex, reason for consultation, source of referral, findings during upper airway examination, age, weight z-score (reflecting how much a finding differs from the mean and in what direction in a normally distributed sample), height z-score, body mass index (BMI) z-score, number of apneas, number of hypopneas, apnea index, hypopnea index, apnea hypopnea index (AHI), oxygen saturation (mean and minimum) measured by pulse oximetry, number of snores, and snore index. RESULTS: Of the 400 patients studied, 242 (60.5%) were male and 158 (39.5%) female. The mean age was 4.95 years. OSAHS (AHI> or =3) was diagnosed in 298 cases (74.5%) and these patients were then studied to determine the relation between OSAHS and obesity. The anthropometric distribution (expressed as mean [SD]) was as follows: weight z score, 0.37 (1.31); height z-score, 0.23 (1.19); BMI, 17.063 kg/m(2) (2.51); and BMI z-score, 0.39 (1.36). The respiratory polygraph during sleep recorded an AHI of 6.56 (7.56). CONCLUSIONS: No differences were observed between the height z score, weight z-score, BMI z-score, age, and AHI. No association between obesity and OSAHS was found in this series. However, studies of larger, unselected populations are needed to determine if obesity is a risk factor for OSAHS in children. PMID- 17125692 TI - [Validation study of the Spanish adaptation of the satisfaction with inhaled asthma treatment questionnaire]. AB - OBJECTIVE: Although satisfaction with asthma treatment is an important variable, it has received little attention and few validated measurement instruments are available. The aim of this study was to assess the psychometric properties and the feasibility of the Spanish adaptation of the Satisfaction With Asthma Treatment Questionnaire (SATQ) for inhaled medication. PATIENTS AND METHODS: This cross-sectional study to validate the adapted SATQ included 239 patients (67.8% women) with stable asthma (mean age, 43.3 years; forced expiratory volume in 1 second, 89% of predicted) and a median time since diagnosis of 10 years. Intermittent asthma was reported in 23.0% of the patients and severe persistent disease in 15.9%. Patients filled out the questionnaire twice. RESULTS: A 4 dimensional factorial structure could be discerned. These dimensions were similar to the validation of the original questionnaire and explained 55.1% of the variance. Internal consistency was acceptable (Cronbach alpha, 0.880 for the overall score and 0.600-0.866 for subscales), as was reliability (intraclass correlation coefficient, 0.912 for the overall score and 0.841-0.916 for subscales). A significant moderate correlation was observed between the overall score and physician--and patient--reported satisfaction (rho=0.295 and 0.507, respectively). Three of the 4 subscales were able to discriminate between categories of the Global Initiative for Asthma. The questionnaire was completed in less than 10 minutes by 92.5% of the patients, and 93.7% left no field blank. CONCLUSIONS: The Spanish adaptation of the SATQ is a useful, user-friendly instrument for measuring satisfaction with inhaled treatment. PMID- 17125691 TI - [Lung cancer diagnosis: hospitalization costs]. AB - OBJECTIVE: To establish the direct costs of the process of diagnosing lung cancer in 2003. As a secondary objective, the cost of admissions defined as inappropriate was evaluated. MATERIAL AND METHODS: A prospective cohort study of lung cancer cases diagnosed in 2003 was performed. Diagnosis was based on cytohistology or clinical and radiological criteria. The total cost was determined according to Decree 222/2003, governing Galician health service rates. A distinction was drawn between hospitalized patients and outpatients, and between small cell and non-small cell carcinomas. Inappropriate admissions were analyzed in accordance with the criteria established by our study team, and the savings that would have been made had these patients been treated as outpatients were calculated. The statistical analyses were performed using SPSS version 10.0. RESULTS: A total of 160 patients were diagnosed with lung cancer; 76 (47.5%) of these were outpatients, and the remaining 84 (52.5%) were hospitalized patients. Admissions were considered inappropriate in 27 cases. Of the total of 160 patients, 108 were diagnosed as having non-small cell carcinomas, and 38 as having small cell carcinomas; the remaining 14 patients were diagnosed on the basis of clinical-radiological criteria. Total cost was 742,847 Euro(mean, 4643 Euro; 95% confidence interval, 4049-5236 Euro), composed of 552,614 Euro(mean, 6579 Euro) for admitted patients, and 190,233 Euro(mean, 2503 Euro) for outpatients. Mean cost was 3692 Euro for the small cell carcinomas, and 5070 Euro for the non-small cell carcinomas. Comparing limited and extensive small cell carcinomas, the mean cost for the former was significantly lower than for the latter (1894 Euro compared to 4098 Euro); there was also a lower mean cost for early compared to advanced stages of non-small cell carcinomas (3660 Euro compared to 5494 Euro). The savings to be made from unnecessary admissions were calculated at 120,258 Euro. CONCLUSIONS: The mean cost for outpatient lung cancer treatment was 62% lower than for hospitalization. Non-small cell carcinomas were more costly on average than small cell carcinomas, and advanced stages of the small cell carcinomas involved a higher average cost than the initial stages of the disease. For our series, the savings to be made from unnecessary admissions were calculated at 120,258 Euro. PMID- 17125695 TI - [Epidemiology of lung cancer in Spain and forecast for the future]. AB - Lung cancer, a steadily growing problem, ranks as the first cause of tumor related deaths in developed countries. The relation between lung cancer and smoking makes it a potentially avoidable disease. Found mainly in men, it has made alarming gains among women. The main prognostic factor is the possibility of receiving curative surgery; however, in real practice the diagnosis usually comes when the disease has reached an advanced stage, when only 20% can be treated surgically. Nonsurgical treatments based on chemo- and radiotherapy have not advanced appreciably in recent years, and 5-year survival is poor, estimated at only around 7% to 12% in Spain. Attempts must be made to improve preventive measures and early diagnosis in order to improve the prognosis for lung cancer patients. PMID- 17125697 TI - [Domiciliary respiratory muscle training in myotonic dystrophy]. AB - A 42-year-old man diagnosed with myotonic dystrophy experienced loss of respiratory muscle strength over a period of 6 months. We report the application of a domiciliary training program targeting both inspiratory and expiratory muscles. Maximal inspiratory and expiratory pressures, forced vital capacity, and forced midexpiratory flow rate were measured 6 months before start of training, just before commencement of the program, and immediately after 12 weeks of training. Adherence to the program was satisfactory. Inspiratory muscle training was efficacious in increasing respiratory muscle strength. Expiratory muscle training, which made use of the Threshold PEP bronchial hygiene device incorporating an adapted flutter valve, was not efficacious in increasing maximal expiratory pressure or halting its loss. However, decreased obstruction of medium caliber airways was observed with use of the device. PMID- 17125694 TI - [Health-related quality of life of patients receiving home mechanical ventilation: the Spanish version of the severe respiratory insufficiency questionnaire]. AB - OBJECTIVE: Home mechanical ventilation is used with patients with severe, chronic respiratory failure, a condition that has a serious impact on quality of life. The aim of this study was to produce a translation and cultural adaptation of the Severe Respiratory Insufficiency. Questionnaire for the Spanish population, the first health-related quality of life questionnaire specifically designed for patients receiving home mechanical ventilation. METHODS: Four bilingual German Spanish translators were used to translate and back-translate the questionnaire. Meetings were held with the translators following each step of the translation process to produce a single version that could be used in the next step. At the end of the process, the questionnaire was piloted to assess its comprehensibility. A scoring system using a scale of 1 (lowest) to 10 (highest) was used to rate both translation difficulty and the naturalness of the language produced. The equivalence of the original and translated items was also evaluated. RESULTS: Three Spanish versions of the questionnaire were produced. Task difficulty was rated as quite low: the mean (SD) ratings were 1.4 (0.6) for translation and 2.2 (1.1) for back translation. The naturalness of the translated items was rated as very high, with scores improving with the successive versions (version 1, 8.4; version 2, 8.7; version 3, 9.1; P< .001). Thirty of the questionnaire items (61.2%) were judged to be fully equivalent, 13 (26.5%) to be similar, and 6 (12.2%) to be non-equivalent. The meaning conveyed by 5 of the items was changed or clarified during piloting. CONCLUSIONS: The translation of the questionnaire using the translation-back-translation procedure has produced a version that is both comparable to the original and accessible to the Spanish population. Its validity is currently being tested in a multicenter study. PMID- 17125696 TI - [Tobacco addiction: care and services]. PMID- 17125698 TI - [Constrictive pericarditis as the first sign of lung cancer]. AB - Symptoms such as cough and hemoptysis in patients with lung cancer can be the consequence of local bronchopulmonary disease, tumor growth that leads to compression of surrounding structures, distant metastases, diverse systemic effects (anorexia, asthenia, weight loss), or paraneoplastic syndromes associated with tumor production of certain hormones. Approximately 10% of patients are asymptomatic at diagnosis. We report the case of a 77-year-old man with dyspnea, pleuritic chest pain, and lower limb edema. The patient died within a few days. The cause of the clinical picture was constrictive pericarditis secondary to metastases from lung carcinoma. PMID- 17125690 TI - [The shuttle run test is not valid for the detection of asthma in school physical education programs]. AB - OBJECTIVE: Certain sporting activities may trigger asthma exacerbations of varying intensity in children. Such exacerbations may lead to limitations in and rejection of such activities. During school hours, teachers are in a good position to observe these phenomena. The aim of the present study was to evaluate the shuttle run, a test of physical fitness used in school physical education programs, as a way of detecting asthma. PATIENTS AND METHODS: We carried out a cross-sectional observational study of school children between the ages of 6 and 12 years using the asthma symptom questionnaire of the International Study of Asthma and Allergies in Childhood (ISAAC), a shuttle run test, and a free running test at maximum effort in order to study bronchial hyperresponsiveness. The principal measure of bronchial hyperresponsiveness used in both physical fitness tests was peak expiratory flow rate as measured with a peak flow meter. In comparing the results of the shuttle run test with those of the free running test and the ISAAC questionnaire we used the chi(2) test to measure association and the Cohen kappa coefficient to measure agreement. RESULTS: We distributed the ISAAC questionnaire (n=919) to 460 (50.1%) boys and 459 (49.9%) girls between the ages of 6 and 12 years. All the tests were completed by 826 children. The level of agreement between the shuttle run test and free running test was positive but low for decreases in peak expiratory flow rate compared to baseline of 15% (chi(2)=5.6; P< .05; kappa=0.093; SE, 0.042) and of 20% (chi(2)=4.5; P< .05; kappa=0.08; SE, 0.046). For 10% decreases association was not significant and agreement was low (kappa=0.05; SE, 0.04). There was no agreement between the ISAAC questionnaire and the shuttle run test (kappa=0.095; SE, 0.63). CONCLUSIONS: The shuttle run test using peak expiratory flow rate as the principal measure of bronchial hyperresponsiveness is not valid for the detection of asthma in schoolchildren. PMID- 17125699 TI - [Necrotizing pneumonitis due to fistulation from the esophageal diverticulum]. PMID- 17125700 TI - [Can we modify bronchial remodeling in asthma?]. PMID- 17125701 TI - [Autoimmunity in chronic obstructive pulmonary disease (COPD)]. PMID- 17125702 TI - [Idiopathic pulmonary fibrosis: in search of an effective treatment]. PMID- 17125703 TI - [Genetic variability in susceptibility to and severity of pneumonia]. PMID- 17125704 TI - [Cardiovascular risk in obstructive sleep apnea-hypopnea syndrome (OSA/HS)]. PMID- 17125705 TI - [The patient with chronic obstructive pulmonary disease in the intensive care unit]. PMID- 17125706 TI - [A friend in need is a friend indeed: the case of drug-eluting stents, diabetes and small vessel size]. PMID- 17125707 TI - [Hypertrophic cardiomyopathy: never-ending complexity]. PMID- 17125708 TI - [Utility of the effective regurgitant orifice for the quantification of regurgitant valve lesions]. PMID- 17125709 TI - [Efficacy of sirolimus-eluting stent implantation in diabetic patients with very small vessels (< or = 2.25 mm). Insights from the DIABETES trial]. AB - INTRODUCTION AND OBJECTIVES: Diabetic patients frequently have small-diameter vessels, which increases their risk of restenosis. The aim of this study was to determine the efficacy of sirolimus-eluting stent implantation in these high-risk patients following percutaneous coronary intervention. METHODS: Our study population comprised a subset of 85 diabetic patients from the DIABETES (DIABETes and sirolimus Eluting Stent) trial who had very small vessels, defined as those with a reference diameter < or =2.25 mm. In the 100 lesions treated, 49 sirolimus eluting stents and 51 bare-metal stents were used. Glycoprotein IIb/IIIa inhibitors were used as recommended by the protocol and dual antiplatelet therapy was administered for 1 year. RESULTS: Baseline clinical and angiographic characteristics were comparable in the two groups. The patients' mean age was 66 (9) years, 42% were women, and 37% were insulin-dependent. On average, the lesion length was 15.0 (9.0) mm and the reference diameter was 1.9 (0.2) mm. At 9-month follow-up, both late lumen loss and the restenosis rate were significantly lower in the sirolimus-eluting stent group than in the bare-metal stent group, at -0.03 (0.3) mm vs 0.44 (0.5) mm (P< .001), and 9.1% vs 39.1% (P=.001), respectively. These differences were also observed in the subgroup of insulin-dependent patients. At 1-year follow-up, the stent thrombosis rate was 0% in the sirolimus eluting stent group, whereas two patients in the bare-metal stent group presented with stent thrombosis. CONCLUSIONS: Sirolimus-eluting stent implantation in diabetics with very small vessels is safe and effective, even in insulin dependent patients. PMID- 17125710 TI - [Beta-myosin heavy-chain gene mutations in patients with hypertrophic cardiomyopathy]. AB - INTRODUCTION AND OBJECTIVES: To determine the frequency of mutations in the beta myosin heavy-chain gene (MYH7) in a cohort of patients with hypertrophic cardiomyopathy (HCM) and their families, and to investigate correlations between genotype and phenotype. METHODS: Single-strand conformation polymorphism analysis and sequencing of fragments with abnormal MYH7 gene mobility were carried out in 128 consecutive index patients with HCM. The phenotypes of patients with and without mutations were compared and the phenotypes of identified families were recorded. RESULTS: A total of 11 mutations were found in 13 families (10%); 7/11 had been previously described. The I736T mutation was found in three families and the A797T in two. One patient had two mutations (i.e., I736T and R787H). Mutations were more frequent in patients with a family history of sudden death (31%) and in those with severe hypertrophy (39% had a thickness > or = 30 mm). Mutations were found in 29 of 42 members of the 13 families, including six family members (20%) who were healthy carriers and aged < or = 36 years. Sudden death had occurred in eight members of four families: four in two families with the I736T mutation, one in a family with A797T, one in a family with R870H, and two in a family with A901P. CONCLUSIONS: MYH7 mutations were present in 10% of our families. Mutations were more frequent in patients with a family history of sudden death and in those with severe hypertrophy. Most mutations had been described previously. Some appeared in several families. For some mutations, the correlation between genotype and phenotype was stable, while for others, there were marked differences between the phenotypes of the index patients and their relatives, suggesting the presence of additional genetic factors that have yet to be identified. PMID- 17125711 TI - [Evaluation of mitral regurgitation severity using a simplified method based on proximal flow convergence]. AB - INTRODUCTION AND OBJECTIVES: Calculation of the effective regurgitant orifice (ERO) is regarded as the most accurate way of assessing the severity of mitral regurgitation (MR), but the technique's complexity limits its use. Our objective was to modify and validate a previously published semiquantitative method of assessment based on measurement of the proximal isovelocity surface area (PISA) in order to adapt it to recent recommendations from American and European cardiology societies. METHODS: In the PISA method, maximum regurgitant flow (MRF) is a function of the radius and aliasing velocity (AV). Using this relationship, it is possible to construct a nomogram formed by lines of different MRF value, which can be easily derived by looking for radius values on the graph and observing where they cross with AV values. The MR severity limits on the nomogram were set to reflect the different severity grades and limits recommended for use with ERO measurements by American and European cardiology societies. RESULTS: We studied 76 patients with MR using Doppler echocardiography. There was an excellent correlation between MRF and ERO (r=0.98, P< .001). Estimates of MR severity made using the new nomogram were in good agreement with those derived from the ERO: for a scale with three severity grades, kappa was 0.951 and the standard error was 0.11; for four grades, kappa was 0.969 and the standard error, 0.11. CONCLUSIONS: Estimates of MR severity derived semiquantitatively from MRF using the nomogram proposed here were in excellent agreement with quantitative estimates obtained using the ERO, and the method was faster and easier to use. PMID- 17125712 TI - [Coronary disease risk and prevalence of heart disease in primary care patients with hypertension and renal disease]. AB - INTRODUCTION AND OBJECTIVES: The presence of renal disease significantly alters the cardiovascular risk of patients with high blood pressure. However, few studies have examined renal parameters in primary care patients. The objectives of this study were to investigate cardiovascular risk and the factors influencing it in hypertensive patients with renal disease and to compare the findings with those in hypertensive patients without renal disease. METHODS: The CORONARIA study involved primary care patients with hypertension from all regions of Spain and included two groups with different degrees of renal disease. RESULTS: In total, 703 patients (9.8%) had renal disease. Hypertensive patients with renal disease had a worse cardiovascular risk profile than other hypertensives. The prevalence of diabetes was double in patients with renal disease. Moreover, the risk of a coronary event was significantly higher in those with renal disease. One-third of hypertensives with renal disease had another previously diagnosed cardiovascular disease. In addition, they more frequently had a history of angina, were twice as likely to have had a myocardial infarction, and were more than twice as likely to have undergone revascularization or to have peripheral vascular disease or cerebrovascular disease. Heart failure was four times more frequent in these patients with renal disease than in other hypertensives. CONCLUSIONS: Patients with hypertension and renal disease have a higher risk of cardiovascular disease, exhibit an increased prevalence of diabetes, and suffer from more extensive target organ damage. PMID- 17125713 TI - [Impact of diabetes mellitus on heart transplant recipients]. AB - INTRODUCTION AND OBJECTIVES: At present, there is some controversy about the impact of diabetes mellitus on heart transplant patients. The effect of the disease on mortality and on other complications, such as infection or rejection, is unclear. The objective of this study was to investigate these factors in our heart transplant patients. METHODS: We studied 365 consecutive patients who underwent heart transplantation between November 1987 and May 2003. We divided them in three groups according to whether they had pretransplantation diabetes (group 1), de novo diabetes (group 2), or no diabetes (group 3). Baseline variables and the development of complications were recorded, and findings were analyzed using Student's t test, chi squared test, and Kaplan-Meier survival analysis. RESULTS: There was no difference in the 1-year or 5-year survival rate between the groups (P=.24 and P=.32, respectively). Patients with pretransplantation and de novo diabetes were older (54.6 years vs 54.9 years vs 50.6 years, P=.04), had a higher prevalence of hypertension (48% vs 36% vs 23%, P=.001), and had more frequently been treated with tacrolimus (10% vs 12% vs 4%, P=.04) or steroids (92% vs 86% vs 70%, P=.001). The incidence of rejection during follow-up was greater in these two groups (64% vs 70% vs 45%, P=.001). CONCLUSIONS: Neither pretransplantation diabetes nor de novo diabetes had a negative impact on survival in our heart transplant patients. The disease's presence was associated with treatment with steroids and tacrolimus. In these patients it would be preferable to individualize immunosuppressive therapy. PMID- 17125714 TI - [Therapeutic success of a prospective cardioversion protocol for persistent atrial fibrillation]. AB - INTRODUCTION AND OBJECTIVES: The best therapeutic approach for persistent atrial fibrillation has yet to be defined. Our aim was to investigate the effects of cardioversion in unselected patients with persistent atrial fibrillation who were treated according to a strict protocol involving pretreatment, cardioversion, and follow-up. METHODS: Consecutive patients with persistent atrial fibrillation of at least 1 months' duration were included prospectively in a cardioversion protocol that involved standard antiarrhythmic pretreatment, with amiodarone being offered first, and follow-up. RESULTS: The study included 295 patients, 87.5% of whom were taking the antiarrhythmic drug amiodarone. Sinus rhythm was restored in 92.5%, with pharmacologic cardioversion occurring in 9.5%. The recurrence rate was 33.5% in the first month and 54.9% by month 12. Antiarrhythmic treatment had to be modified in 10.8% of patients. Independent risk factors for recurrence during the first year after cardioversion were an atrial fibrillation duration greater than one year, previous cardioversion, and left ventricular dilatation. A simple risk scoring system was able to differentiate between subgroups of patients with a low, intermediate or high risk of recurrence in the first year after cardioversion. CONCLUSIONS: Sinus rhythm was maintained for 1 year after effective cardioversion in 45.1% of patients who received homogeneous antiarrhythmic pretreatment. There were few side effects. Recurrence can be predicted using clinical variables such as left ventricular dilatation, arrhythmia duration, and previous cardioversion. PMID- 17125715 TI - [Altered fibrillar collagen metabolism in hypertensive heart failure. Current understanding and future prospects]. AB - Arterial hypertension induces numerous alterations in the composition of cardiac tissue, which, in turn, result in structural remodeling of the myocardium. This remodeling is due to a range of pathologic mechanisms associated with mechanical, neurohormonal and cytokine processes that affect both cardiomyocyte and non cardiomyocyte compartments of the myocardium. One of these processes involves disruption of the equilibrium between the synthesis and degradation of type-I and type-III collagen molecules. The result is excess accumulation of type-I and type III collagen fibers in interstitial and perivascular spaces in the myocardium. The clinical significance of myocardial fibrosis lies in its contribution to the development of cardiac complications in hypertensive patients. This brief review focuses on the mechanisms of myocardial fibrosis and their clinical consequences. In addition, the techniques used for diagnosing myocardial fibrosis and the main therapeutic strategies for reducing fibrosis are also discussed. PMID- 17125717 TI - [Multidetector computed tomography of the coronary veins and resynchronization therapy]. PMID- 17125716 TI - [Menopausal hormone therapy and cardiovascular disease]. AB - Despite biologically plausible mechanisms for cardiac protection and compelling evidence from observational studies suggesting that menopausal hormone therapy confers cardiovascular benefit, results of well-designed and conducted randomized clinical trials in healthy women and in women with established coronary heart disease displayed that menopausal hormone therapy failed to prevent clinical cardiovascular events and rather was associated with harms. Clinical trial of the SERM raloxifene also did not demonstrate a decrease in coronary events. It is unknown whether the earlier initiation of such therapies, i.e., at menopause, would result in favorable outcomes; or whether different hormonal preparations, lower doses, or alternate routes of administration would confer benefit. At present, proved coronary risk reduction strategies are requisite (albeit underutilized) for menopausal women; these include lifestyle and pharmacologic coronary preventive interventions. The baseline characteristics of menopausal women with coronary heart disease who were participants in cardiovascular outcome trials of menopausal hormone therapy or raloxifene were remarkably similar; globally, cardiovascular risk factors were not optimally controlled at entry into these trials, suggesting that more aggressive cardiovascular risk interventions are appropriate to achieve optimal target goals for menopausal women with documented coronary heart disease. PMID- 17125718 TI - [Right versus left radial artery approach for coronary angiography. Differences observed and the learning curve]. AB - There are anatomical differences between right and left radial artery approaches for coronary catheterization that could influence application of the technique. We present the results of a randomized study that compared the effectiveness of the two approaches and identified factors associated with failure of the procedure. The study involved 351 consecutive patients: a left radial approach was used in 180, and a right radial approach, in 171. The procedure could not be completed using the initial approach selected in 15 patients (11 right radial vs. 4 left radial; P=.007). Use of a right radial approach, lack of catheterization experience, patient age >70 years, and the absence of hypertension were found to be independently associated with prolonged fluoroscopy duration and failure using the initial approach. Use of the right radial approach in patients aged over 70 years was associated with a 6-fold increase in the risk of an adverse event. Consequently, use of the right radial approach should be avoided in patients aged over 70 years when trainee practitioners are on the learning curve. PMID- 17125719 TI - [Stability of NT-ProBNP in patients with systolic heart failure]. AB - Natriuretic peptides are extremely useful in the diagnosis and prognosis of patients with heart failure. However, it is not clear whether their values are stable. We carried out a prospective study of 30 consecutive ambulatory patients (mean age, 62.6 [12.2] years) with stable systolic heart failure, as determined by the 6-minute walk test, who were in New York Heart Association class II or III and who had a left ventricular ejection fraction <30% (mean ejection fraction, 24.2% [6.68%]). At baseline, the mean N-terminal pro-brain natriuretic peptide (NT-proBNP) level and the mean distance walked in 6 minutes were 2237.3 pg/mL and 348.26 m, respectively. At 3-month follow-up, the corresponding values were 2096.2 pg/mL and 372.05 m, respectively. No significant difference was observed in NT-proBNP level or in distance walked in 6 minutes between baseline and 3 months (P=.8). Overall, there was a good correlation (r=0.94; P< .001) between the plasma NT-proBNP level at baseline and at 3 months in patients with stable chronic heart failure due to systolic dysfunction in New York Heart Association class II or III. PMID- 17125720 TI - [Arrhythmic storm induced by AICD discharge in a patient with catecholaminergic polymorphic ventricular tachycardia]. PMID- 17125721 TI - [A case of idiopathic effusive-constrictive pericarditis]. PMID- 17125722 TI - [Pericardial cyst: all's well that ends well]. PMID- 17125723 TI - Effectiveness and benefit-cost of peer-based workplace substance abuse prevention coupled with random testing. AB - Few studies have evaluated the impact of workplace substance abuse prevention programs on occupational injury, despite this being a justification for these programs. This paper estimates the effectiveness and benefit-cost ratio of a peer based substance abuse prevention program at a U.S. transportation company, implemented in phases from 1988 to 1990. The program focuses on changing workplace attitudes toward on-the-job substance use in addition to training workers to recognize and intervene with coworkers who have a problem. The program was strengthened by federally mandated random drug and alcohol testing (implemented, respectively, in 1990 and 1994). With time-series analysis, we analyzed the association of monthly injury rates and costs with phased program implementation, controlling for industry injury trend. The combination of the peer-based program and testing was associated with an approximate one-third reduction in injury rate, avoiding an estimated $48 million in employer costs in 1999. That year, the peer-based program cost the company $35 and testing cost another $35 per employee. The program avoided an estimated $1850 in employer injury costs per employee in 1999, corresponding to a benefit-cost ratio of 26:1. The findings suggest that peer-based programs buttressed by random testing can be cost-effective in the workplace. PMID- 17125724 TI - Reversible covalent inhibition of a phenol sulfotransferase by coenzyme A. AB - Phenol sulfotransferases (SULTs), which normally bind 3'-phosphoadenosine-5' phosphosulfate as the donor substrate, are inhibited by CoA and its thioesters. Here, we report that inhibition of bovine SULT1A1 by CoA is time-dependent at neutral pH under non-reducing conditions. The rates of inactivation by CoA indicate an initial reversible SULT:CoA complex with a dissociation constant of 5.7 microM and an inactivation rate constant of 0.07 min(-1). Titrations with CoA and prolonged incubations reveal that inactivation of the dimeric enzyme is stoichiometric, consistent with the observation of complete conversion of the protein to a slightly decreased electrophoretic mobility. Both activity and normal electrophoretic migration are restored by 2-mercaptoethanol. Mutagenesis demonstrated that Cys168 is the site of CoA adduction, and a consistent model was constructed that reveals a new SULT molecular dynamic. Cysteine reaction kinetics with Ellman's reagent revealed a PAPS-induced structural change consistent with the model that accounts for binding of CoA. PMID- 17125725 TI - Voltage-dependent modulation of L-type calcium currents by intracellular magnesium in rat ventricular myocytes. AB - Effects of changing cytosolic free Mg(2+) concentration on L-type Ca(2+) (I(Ca)) and Ba(2+) currents (I(Ba)) were investigated in rat ventricular myocytes voltage clamped with pipettes containing 0.2 or 1.8mM [Mg(2+)] ([Mg(2+)](p)) buffered with 30mM citrate and 10mM ATP. Increasing [Mg(2+)](p) from 0.2 to 1.8mM reduced current amplitude and accelerated its decay under a variety of experimental conditions. To investigate the mechanism for these effects, steady-state and instantaneous current-voltage relationships were studied with two-pulse and tail current (I(T)) protocols, respectively. Increasing [Mg(2+)](p) shifted the V(M) for half inactivation by -20mV but dramatically decreased I(Ca) amplitude at all potentials tested, consistent with a change in gating kinetics that decreases channel availability. This conclusion was supported by analysis of I(T) amplitude, but these latter experiments also suggested that, in the millimolar concentration range, [Mg(2+)](p) might also inhibit permeation through open Ca(2+) channels at positive V(M). PMID- 17125726 TI - Lithium ions increase hepatic glycogen synthase stability through a proteasome related mechanism. AB - Incubation of rat hepatocytes with LiCl resulted in an overall increase in the activity ratio of glycogen synthase (GS), concomitantly with a decrease in active GS kinase-3 levels. GS total activity was also increased in a dose- and time dependent manner. This latter effect correlated with the amount of immunoreactive enzyme determined by immunoblotting. Cycloheximide and actinomycin-D did not modify LiCl action on GS activity. Lithium ions did not induce any changes in GS mRNA levels. Furthermore, the increase in the total amount of GS induced by LiCl was further augmented after addition of a specific, calpain and proteasome inhibitor. Our results indicate that LiCl increases hepatocyte GS activity through increasing both the activation state of the enzyme and its cellular content. This latter increase is mediated through a modification of the proteasome-regulated proteolytic pathway of the enzyme. PMID- 17125727 TI - Strong cardioprotective effect of resveratrol, a red wine polyphenol, on isolated rat hearts after ischemia/reperfusion injury. AB - We have studied some hemodynamic parameters as heart rate (HR) developed pressure (DP) and maximal positive values of the first derivative of pressure (+dP/dt max) in isolated heart from control or resveratrol treated rats. In acute ex vivo experiments, resveratrol (1-100 microM) infusion in Langendorff perfused hearts did not affect contractile function in either normoxic conditions or after ischemia/reperfusion. However when semi-chronically administered by IP injection during 7 days, resveratrol which had no effect on pre-ischemic heart greatly improved post-ischemic indexes of myocardial function. Resveratrol effect is dose dependent and seemed optimal at a plasma level of 18.5 microM. This concentration is very close to that previously shown to be optimal and non-toxic by others. These beneficial effects of resveratrol are only partly explained by its antioxidant properties as suggested by the lack of any dose-response effect on tissue malondialdehyde (MDA) levels. They are also clearly not mediated by nitric oxide (NO) elevation. When acutely infused resveratrol had no beneficial effect and therefore could not be proposed in acute scenarios of ischemia/reperfusion or stroke. However resveratrol appeared as an efficient and promising molecule in the prevention of heart dysfunction. PMID- 17125728 TI - Phenotype and enamel ultrastructure characteristics in patients with ENAM gene mutations g.13185-13186insAG and 8344delG. AB - OBJECTIVE: The main clinical manifestations of amelogenesis imperfecta (AI) include alteration in the quality and quantity of enamel. AI is associated with different mutations in four genes: enamelin (ENAM), amelogenin (AMGX), kallikrein (KLK4) and enamelysin (MMP-20). Seven different mutations have been identified in the enamelin gene (ENAM). DESIGN: In this paper, we describe the phenotype and ultrastructure of enamel observed using scanning electron microscopy (SEM) in patients with two autosomal dominant (AD) mutations in the ENAM gene: g.13185 13186insAG and g.8344delG, each in one of two unrelated families. Mutations were confirmed by sequence analysis of PCR amplified products of all 10 exons and exon/intron boundaries of the ENAM gene. RESULTS: Phenotypic diversity was observed in patients with ENAM gene mutations g.13185-13186insAG with consecutive protein alteration designated as p.P422fsX488 within family 1. In the proband, the enamel of his entire dentition was chalky white with only mild local hypoplastic alteration, while the phenotypic appearance of his father's dentition was that of local hypoplastic AI. In patients with the ENAM gene mutation g.8344delG from family 2 with consecutive protein alteration designated as p.N197fsX277, generalised hypoplastic AI was observed. CONCLUSIONS: Ultrastructural enamel changes in the patient with the autosomal dominant ENAM g.13185-13186insAG mutation, described for the first time in this study, were less pronounced compared to ultrastructural changes in patients with the autosomal dominant ENAM mutation 8344delG. Ultrastructural characteristics of the g.13185-13186insAG mutation revealed deformed prisms, an oval shape on the cross section and wider interprism spaces, while enamel with the ENAM mutation 8344delG was laminated, but prismless. PMID- 17125730 TI - Gland size estimation and body mass index improve salivary flow rate assessment. PMID- 17125729 TI - Age-dependent increase of discoidin domain receptor 2 and matrix metalloproteinase 13 expression in temporomandibular joint cartilage of type IX and type XI collagen-deficient mice. AB - Our previous studies demonstrated that mutations in type IX and type XI collagens in mice caused osteoarthritis (OA)-like changes in knee and temporomandibular (TM) joints. We also found that the overexpression of matrix metalloproteinase 13 (Mmp-13) was probably due to the up-regulation of a collagen receptor, discoidin domain receptor 2 (Ddr2), which was responsible for knee cartilage degeneration in mutant mice. The objective of our study was to determine whether the expression of Mmp-3, Mmp-13 and Ddr2 was increased in OA-like TM joints in mutant mice using immunohistochemistry. We found that the staining for Ddr2, Mmp-13 and Mmp-derived type II collagen fragments in tissue sections from 6-month-old mice was increased in TM joints of the mutant mice. In contrast, we found no difference in the staining for Mmp-3 amongst the two mutant mice and their wild type littermates. We conclude that, similar to previous observations in knee joints, the overexpression of Ddr2 and Mmp-13 may be responsible for the OA-like change in TM joints in mutant mice. PMID- 17125731 TI - Effect of age on alveolar bone turnover adjacent to maxillary molar roots in male rats: A histomorphometric study. AB - OBJECTIVE: The effect of age on alveolar bone turnover adjacent to maxillary molar roots of male rats was assessed histomorphometrically with special focus on bone formation and resorption. DESIGN: A total of 110 male Wistar rats ranging in age from 6 to 100 weeks were used for this study. Histomorphometric parameters were measured in fluorescence-labeled undecalcified ground and paraffin-embedded decalcified sections of the alveolar wall around the disto-lingual roots of the maxillary first molars. Bone formation was measured statically by determining the percentage of the bone surface that was double-labeled surface (dLS/BS), bone formation rate (BFR/BS), and mineral apposition rate (MAR). Bone resorption was quantified statically in terms of the number of osteoclasts (N.Oc/BS) and the percentage of the bone surface covered with osteoclasts (Oc.S/BS). RESULTS: For the total surface of the alveolar wall, the values obtained for all parameters of both bone formation and resorption decreased with advancing age. All these values rapidly decreased during the early part of the life span, from 6 to 30-40 weeks of age, of the rats. A site-specific difference between the distal and mesial sides of the alveolar wall was found for each age group. dLS/BS and BFR/BS were significantly greater (p < 0.0001) on the mesial side than on the distal one. On the other hand, the distal side showed significantly greater (p < 0.0001) value for N.Oc/BS and Oc.S/BS did the mesial one. However, there were no significant age-related changes in dLS/BS and BFR/BS on the distal side or in N.Oc/BS and Oc.S/BS on the mesial side throughout observation period. CONCLUSION: The results of the present study demonstrate that alveolar bone turnover of male rats decreased rapidly with advancing age but that in order to maintain the integrity of the tooth function mechanical stress may still have participated in bone formation and resorption of the alveolar wall even in rats 100-week old. PMID- 17125732 TI - Dental implications of some common movement disorders: a concise review. AB - Movement disorders - or dyskinesias - are characterized by involuntary movements. Despite the major role for medical specialists in the diagnosis and treatment of dyskinesias, dentists are confronted with such disorders as well. Unfortunately, the literature regarding the dental implications of movement disorders is still scarce. This concise review describes the dental implications of some common dyskinesias, viz., Gilles de la Tourette's syndrome, Huntington's disease, idiopathic torsion dystonia, oral dyskinesias, and Parkinson's disease. It was concluded that these dyskinesias may have profound dental implications. Not only do generalized dyskinesias have focal manifestations in the orofacial region, but there are also dyskinesias that exclusively affect the orofacial area. The oral manifestations of dyskinesias are in part directly related to the disorder, and in part medicine-related. Dentists should be able to recognize the oral manifestations and, when properly trained, to manage them adequately. In most instances, a multidisciplinary approach upon referral is necessary, including the medical specialists involved. Unfortunately, the level of evidence of the selected papers was generally low. In our rapidly ageing population, it is a challenge for all of us to improve the quality of this emerging field, for the sake of this sometimes heavily infirmed category of patients. PMID- 17125733 TI - Does isoprene protect plant membranes from thermal shock? A molecular dynamics study. AB - The question of why plants release isoprene when heat stressed has been continuously debated for more than half a century. In this work we use molecular dynamics simulation techniques to directly investigate the interaction between isoprene and a model phospholipid membrane in atomic detail. It is found that isoprene partitions preferentially in the center of the membrane and in a dose dependent manner enhances the order within the membrane without significantly changing the dynamical properties of the system. At a concentration of 20 mol% isoprene (16 isoprene molecules per 64 lipid molecules) the effect of the addition of isoprene on the membrane order is equivalent to a reduction in temperature of 10 K, rising to a reduction of 30 K at 43 mol% isoprene. The significance of the work is that it provides for the first time direct evidence that isoprene stabilizes lipid membranes and reduces the likelihood of a phospholipid membrane undergoing a heat induced phase transition. Furthermore it provides a clear mechanistic picture as to why plants specifically utilize isoprene for this purpose. PMID- 17125734 TI - Beyond fear and disgust: the role of (automatic) contamination-related associations in spider phobia. AB - This study explored the role of threat and contamination-related associations in spider phobia. Treatment-seeking (n=60) and non-phobic (n=30) individuals completed threat and disgust-related Implicit Association Tests (IATs). Phobic individuals were assessed before and after one session of 2.5h in vivo exposure. To differentiate actual treatment effects from test-retest effects on the IAT, half of the phobic individuals completed the IAT twice before treatment. Results showed that: (1) threat and contamination associations similarly distinguished between phobic and non-phobic participants on self-reports and IATs; (2) only self-reported threat associations incrementally predicted participants' overt avoidance behavior next to self-reported global affective associations; (3) self reported associations were significantly reduced following treatment; (4) IAT effects showed no significant reduction following treatment, and no evidence was found for an additional treatment-induced change over and above test-retest effects. PMID- 17125735 TI - In vivo gene transfer into the honeybee using a nucleopolyhedrovirus vector. AB - The honeybee Apis mellifera L. is a social insect and one of the most industrially important insects. We examined whether a baculovirus-mediated retrotransposon is applicable to in vivo transfer of exogenous genes to the honeybees. Honeybee larvae and pupae were injected with two types of recombinant Autographa californica nucleopolyhedrovirus (AcNPV) vectors, one that includes the enhanced green fluorescent protein gene (egfp) as a reporter to be inserted into the honeybee genome, and another that includes the reverse transcriptase gene responsible for the insertion. Fluorescence was observed in most of the viral-injected larvae and pupae. Reverse transcription-polymerase chain reaction and immunoblotting confirmed egfp mRNA and eGFP expression in these honeybees, although egfp insertion into the honeybee genome was not confirmed. These results indicate that AcNPV vectors can be used for the transfer and transient expression of an exogenous gene in the larval and pupal honeybees. PMID- 17125736 TI - Microarray analysis of gene expression in Men1 knockout embryoid body reveals genetic events involved in early mouse embryonic development. AB - The Men1 gene has been identified as the gene responsible for MEN1, a hereditary syndrome transmitted with an autosomal dominant trait. Disruption of the Men1 gene results in defects of multiple organs development, including the nervous system, heart, liver, cranium, and face. In this study, we used embryoid bodies (EBs) formed from wild-type and Men1-/- ES cells as a model system to investigate effect of Men1 gene on the embryo development. We characterized in detail gene expression profile of these Men1-/- EBs by microarray techniques and identified a series of putative menin targeted genes, including genes involved in development of bone (e.g., Postn, Runx2, and Msx2), liver (e.g., KDR), blood (e.g., Hox9 and Kitl), and pancreatic islet (e.g., Sox4, Foxa1, Btc, Igf2, and Nfatc1). Further studies may shed light onto the underlying mechanisms of the interplay between menin and these genes. PMID- 17125737 TI - Angiogenin-induced protein kinase B/Akt activation is necessary for angiogenesis but is independent of nuclear translocation of angiogenin in HUVE cells. AB - Angiogenin, a potent angiogenic factor, binds to endothelial cells and is endocytosed and rapidly translocated to and concentrated in the nucleolus where it binds to DNA. In this study, we report that angiogenin induces transient phosphorylation of protein kinase B/Akt in cultured human umbilical vein endothelial (HUVE) cells. LY294002 inhibits the angiogenin-induced protein kinase B/Akt activation and also angiogenin-induced cell migration in vitro as well as angiogenesis in chick embryo chorioallantoic membrane in vivo without affecting nuclear translocation of angiogenin in HUVE cells. These results suggest that cross-talk between angiogenin and protein kinase B/Akt signaling pathways is essential for angiogenin-induced angiogenesis in vitro and in vivo, and that angiogenin-induced PKB/Akt activation is independent of nuclear translocation of angiogenin in HUVE cells. PMID- 17125738 TI - Non-hypoxic induction of HIF-3alpha by 2-deoxy-D-glucose and insulin. AB - Hypoxia-inducible factors (HIFs) are key mediators of cellular adaptation to hypoxia, but also respond to non-hypoxic stimuli. To clarify involvement in metabolic disturbances, HIFs were characterised in rats subjected to insulin induced hypoglycaemia or cellular glucoprivation provoked by 2-deoxy-D-glucose (2 DG). Using real-time qPCR, organ-specific expression of HIF-1alpha, -2alpha, 3alpha, -1beta, and of the target gene GLUT-1 was determined. Distribution of HIF 3alpha proteins was examined by immunohistochemistry. Both, insulin and 2-DG resulted in a widespread increase in HIF-3alpha mRNA. HIF-2alpha mRNA increased in lung and heart after 2-DG only, whereas other HIFs remained unaffected. A pronounced increase of protein levels in cerebral cortex was observed for HIF 3alpha. Functional significance of HIF induction was reflected in enhancement of GLUT-1 mRNA. Transcriptional up-regulation of HIF-3alpha represents a typical response to in vivo hypoglycaemia and glucoprivation. These data suggest an involvement of the HIF system in metabolic derangements as for instance caused by diabetes. PMID- 17125739 TI - Berberine alters the processing of Alzheimer's amyloid precursor protein to decrease Abeta secretion. AB - Berberine is an isoquinoline alkaloid isolated from Coptidis rhizoma, a major herb widely used in Chinese herbal medicine. Berberine's biological activity includes antidiarrheal, antimicrobial, and anti-inflammatory effects. Recent findings show that berberine prevents neuronal damage due to ischemia or oxidative stress and that it might act as a novel cholesterol-lowering compound. The accumulation of amyloid-beta peptide (Abeta) derived from amyloid precursor protein (APP) is a triggering event leading to the pathological cascade of Alzheimer's disease (AD); therefore the inhibition of Abeta production should be a rational therapeutic strategy in the prevention and treatment of AD. Here, we report that berberine reduces Abeta levels by modulating APP processing in human neuroglioma H4 cells stably expressing Swedish-type of APP at the range of berberine concentration without cellular toxicity. Our results indicate that berberine would be a promising candidate for the treatment of AD. PMID- 17125740 TI - Uniform amplification of multiple DNAs by emulsion PCR. AB - When several DNAs are amplified by PCR in one PCR tube, biased amplification is known to occur because amplification efficiency differs from one DNA to another. Therefore, we conducted PCR in the water in oil-emulsion (W/O emulsion) to examine whether the procedure allows the uniform amplification of several DNAs. In the amplification of a model library consisting of two clones, the emulsification of the PCR mixture successfully reduced the difference in its amplification efficiency to approximately one-seventh the value obtained without emulsification. Furthermore, we conducted repeated PCR to amplify a model library consisting of ten short hairpin RNA (shRNA) expression vectors as a model experiment for gene discovery using an shRNA expression library. Consequently, the emulsification of the PCR mixture successfully reduced PCR bias. Our results indicate that emulsion PCR is capable of uniformly amplifying libraries of shRNA, ribozyme, cDNA, and others, and is useful also for gene discovery using these libraries. PMID- 17125741 TI - Constitutive activity and inverse agonism at the alpha1adrenoceptors. AB - Mutations of G protein-coupled receptors (GPCR) can increase their constitutive (agonist-independent) activity. Some of these mutations have been artificially introduced by site-directed mutagenesis, others occur spontaneously in human diseases. The alpha(1B)adrenoceptor was the first GPCR in which point mutations were shown to trigger receptor activation. This article briefly summarizes some of the findings reported in the last several years on constitutive activity of the alpha(1)adrenoceptor subtypes, the location where mutations have been found in the receptors, the spontaneous activity of native receptors in recombinant as well as physiological systems. In addition, it will highlight how the analysis of the pharmacological and molecular properties of the constitutively active adrenoceptor mutants provided an important contribution to our understanding of the molecular mechanisms underlying the mechanism of receptor activation and inverse agonism. PMID- 17125742 TI - Testosterone levels and sexual maturation predict substance use disorders in adolescent boys: a prospective study. AB - BACKGROUND: This investigation determined whether testosterone level and sexual maturation in boys biased development of socially nonnormative behavior culminating in a substance use disorder (SUD). METHODS: The subjects were 179 boys recruited in late childhood through a high-risk paradigm. Path analysis was used to evaluate the influence of testosterone level and sexual maturation in early adolescence (age 12-14) on attitudes toward antisociality, affiliation with deviant peers, and social potency in middle adolescence (age 16), illicit drug use by late adolescence (age 19), and SUD in young adulthood (age 22). RESULTS: Testosterone level predicted social potency and approval of aggressive/antisocial behavior. Sexual maturation mediated the relation between testosterone level in early adolescence and later affiliation with deviant peers. Social potency, approval of aggressive/antisocial behavior, and deviant peer affiliations predicted illicit drug use by late adolescence that in turn predicted SUD in young adulthood. CONCLUSIONS: This study demonstrated that pubertal processes in early adolescence influence the risk for SUD via effects on psychosocial functioning. PMID- 17125743 TI - Neonatal handling prevents the effects of phencyclidine in an animal model of negative symptoms of schizophrenia. AB - BACKGROUND: Environmental factors during the neonatal period have long-lasting effects on the brain. Neonatal handling, an early mild stress, enhances the ability to cope with stress in adult rats. In humans, inappropriate stress responses increase the risk of schizophrenia in genetically predisposed individuals. We studied the effect of neonatal handling on the phencyclidine (PCP)-induced immobility time of rats in the forced swimming test (FST, an animal model of negative symptoms of schizophrenia) and on plasma adrenocorticotropic hormone (ACTH) as a measure of hypothalamic-pituitary-adrenal axis (HPA) reactivity. METHODS: Pups were removed from their mothers 15 min/21 days after birth. Postnatal day 65: animals were submitted to restraint stress. Postnatal day 75: after PCP treatment (5 mg/kg/5 days) animals were submitted to the FST. RESULTS: Neonatal handling reduced HPA reactivity to passive stress (restraint) but not to active coping stress (forced swimming). Immobilization time was significantly lower in saline- and PCP-treated, handled animals than in non handled ones. Handling prevented the ACTH increase induced by PCP that was observed in the non-handled rats after FST. CONCLUSIONS: First, neonatal handling protects animals from acquiring the schizophrenic-like behavior provoked by sub chronic PCP treatment, which was associated with a reduced HPA activity. Second, the beneficial properties of handling in stress responses seem to depend on the type of stress. PMID- 17125744 TI - Genetic dissection of the tail suspension test: a mouse model of stress vulnerability and antidepressant response. AB - BACKGROUND: The tail suspension test (TST) is a mouse screening test for antidepressants. METHODS: An F2 intercross was derived from NMRI and 129S6 inbred strains (n = 747). Mice underwent standardized TST with 2 sessions: (1) baseline and (2) imipramine (30 mg/kg, intraperitoneally) TST. RESULTS: A whole genome scan of this intercross mapped significant basal TST quantitative trait loci (QTL) on chromosomes (chr) 5 (peak 61 cM, Lod 5.7), 12 (peak 43 cM, Lod 5.2), and 18 (peak 51 cM, Lod 3.0). A suggestive QTL on chr 4 (peak 62 cM; Lod 3.1) overlapped regions containing previously mapped QTLs. For TST imipramine response, QTL were mapped on chr 1, 4, and 5. The chromosome 5 locus affected basal TST, antidepressant immobility response, and tail suspension-induced hyperthermia (TSIH) behaviors. An outbred NMRI F2 population provided further evidence for a chr 5 QTL. This chr 5 region harbors a cluster of gamma aminobutyric acid (GABA)-A receptor subunits and the human syntenic region includes chr 4p, 1p11, 12q24, and 22q11.24. A significant TSIH QTL (Tsih1) mapped on chr 4 near the Leptin receptor (Lepr). CONCLUSIONS: These QTL provide potential regions of interest for human genetic studies in stress-diathesis models of psychiatric illness and antidepressant responsiveness. PMID- 17125745 TI - Alterations of CREB and DARPP-32 phosphorylation following cocaine and monoaminergic uptake inhibitors. AB - The cAMP response element-binding protein (CREB) is a transcription factor that can contribute to drug-induced changes in gene expression. It is well known that the dopamine and cAMP-regulated phosphoprotein (DARPP-32), via activation, is converted into a potent inhibitor of protein phosphatase-1 (PP-1), which regulates the activity of CREB. We previously reported that the continuous infusion of cocaine for 7 days produced a significant increase in prodynorphin mRNA in the rat caudate putamen and we also studied the role of the different monoamines in these cocaine effects. Since multiple cAMP response element (CRE) sequences are present on the prodynorphin gene promoter, the aim of our study was to investigate the effects of cocaine and monoaminergic uptake inhibitors on CREB and DARPP-32 phosphorylation and moreover the possible correlation with the changes already observed on prodynorphin gene expression. Here we investigated the alterations on phospho-Ser133 CREB, phospho-Thr34 DARPP-32 and phospho-Thr75 DARPP-32 induced by continuous infusions of cocaine, GBR12909, fluoxetine and nisoxetine. A significant decrease in both phospho-CREB at Ser133 and phospho DARPP-32 at Thr34 in the rat caudate putamen was produced by cocaine, GBR 12909, fluoxetine or nisoxetine. No alterations were observed on phospho-Thr75 DARPP-32 levels. We hypothesize that the decrease in phospho-Thr34 DARPP-32 could evoke an increase in PP-1 activity which is responsible for the reduction of CREB activation. These effects could in turn elicit the reduction in the transcriptional cascade of the prodynorphin gene in the caudate putamen, observed following chronic fluoxetine and nisoxetine. On the other hand, these mechanisms do not seem to be involved in cocaine- or GBR 12909-induced effects. PMID- 17125746 TI - Long-term behavioral consequences of brief, repeated neonatal isolation. AB - Rats subjected to stressful stimuli during the stress hyporesponsive period exhibit varied neuroendocrine and behavioral changes as neonates, adolescents and adults. The current work examined the effects of neonatal isolation stress, using a within-litter design, on adult anxiety-related behavior and endocrine stress reactivity. Neonatal rats were isolated daily for 1 h from postnatal day (P) 4 to 9, a manipulation previously shown to induce hypothalamic-pituitary-adrenal (HPA) responses on P9 (Knuth, E.D., Etgen, A.M. (2005) Corticosterone secretion induced by chronic isolation in neonatal rats is sexually dimorphic and accompanied by elevated ACTH. Horm Behav 47:65-75.). Control animals were either handled briefly or left undisturbed (with-dam). Adult rats were tested for anxiety-related behavior using the elevated plus maze and open field, and for endocrine responses following restraint stress. Neonatal isolation decreased center exploration of the open field following 1 h restraint, including decreased time in the center compared to with-dam or handled controls and decreased center entries and distance traveled in the center compared to with-dam controls. It also decreased time in and entries into the open arms of the elevated plus maze compared to handled controls, suggesting enhanced anxiety-related behavior. Neonatal isolation had no effect on basal or restraint-induced levels of ACTH or corticosterone. These findings indicate that neonatal isolation may enhance anxiety-related behaviors, especially in response to stress, without altering HPA function. PMID- 17125747 TI - Effects of intermittent hypoxia on cat petrosal ganglion responses induced by acetylcholine, adenosine 5'-triphosphate and NaCN. AB - Exposure to chronic intermittent hypoxia (CIH) for 4 days enhances the cat carotid body (CB) chemosensory responses to acute hypoxia. However, it is not known if CIH enhances the responses of the petrosal ganglion (PG) neurons that innervate the CB chemoreceptor cells. Accordingly, we studied the effects of the CB putative excitatory transmitter acetylcholine (ACh) and adenosine 5 triphosphate (ATP), and the effects of citotoxic hypoxia (NaCN) applied to the isolated PG from cats exposed to CIH for 4 days. The dose-dependent curve parameters of the frequency of discharges evoked in the carotid sinus nerve by the application of ACh, ATP and NaCN to the isolated PG in control condition were not significantly modified in the CIH-treated cats. Present results suggest that CIH enhances the chemosensory responses to acute hypoxia acting primarily at the chemoreceptor cells, without major changes in the response of PG neurons evoked by the application of putative CB excitatory transmitters to their somata. PMID- 17125748 TI - Vagus nerve stimulation may protect GABAergic neurons following traumatic brain injury in rats: An immunocytochemical study. AB - Seizures and subclinical seizures occur following experimental brain injury in rats and may result from inhibitory neuron loss. This study numerically compares cortical and hippocampal glutamic acid decarboxylase (GAD) positive neurons between sham fluid percussion injury (FPI), FPI with sham Vagus Nerve Simulation (VNS), and FPI with chronic intermittent VNS initiated at 24 h post FPI in rats. Rats (n=8/group) were prepared for immunocytochemistry of GAD at 15 days post FPI. Serial sections were collected and GAD immunoreactive neurons were counted in the hippocampal hilus and two levels of the cerebral cortex. Numbers of quantifiable GAD cells in the rostral cerebral cortices were different between groups, both ipsilateral and contralateral to the FPI. Post hoc analysis of cell counts rostral to the ipsilateral epicenter, revealed a significant 26% reduction in the number of GAD cells/unit area of cerebral cortex following FPI. In the FPI VNS group, this percentage loss was attenuated to only an 8.5% reduction, a value not significantly different from the sham group. In the contralateral side of the rostral cerebral cortex, FPI induced a significant 24% reduction in GAD cells/unit area; whereas, the VNS-treated rats showed no appreciable diminution of GAD cells rostral to the contralateral epicenter. Hippocampal analysis revealed a similar reduction of GAD cells in the FPI group; however, unlike the cortex this was not statistically significant. In the FPI-VNS group, a trend towards increased numbers of hilar GAD cells was observed, even over and above that of the sham FPI group; however, this was also not statistically significant. Together, these data suggest that VNS protects cortical GAD cells from death subsequent to FPI and may increase GAD cell counts in the hippocampal hilus of the injured brain. PMID- 17125749 TI - Mismatch negativity to pitch contours is influenced by language experience. AB - A cross-language study utilizing the mismatch negativity (MMN) evoked response was conducted to explore the influence of language experience on the preattentive cortical processing of linguistically relevant pitch contours. Chinese and English subjects were presented with Mandarin Chinese tones while the mismatch negativity (MMN) response was elicited using a passive oddball paradigm. Two oddball conditions were constructed with a common deviant, a low falling rising contour tone (T3). One condition consisted of two tones that are acoustically similar to one another (T2/T3: T2, high rising contour=standard). The other condition consisted of two tones that are acoustically dissimilar to one another (T1/T3: T1, high level=standard). These tonal pairs enabled us to assess whether different degrees of similarity between pitch movements exert a differential influence on preattentive pitch processing. Results showed that the mean MMN amplitude of the Chinese group was larger than that of the English group for the T1/T3 condition. No group differences were found for the T2/T3 condition. The mean MMN amplitude was larger for the T1/T3 relative to the T2/T3 condition for the Chinese group only. By virtue of these language group differences, we infer that early cortical processing of pitch contours may be shaped by the relative saliency of acoustic dimensions underlying the pitch patterns of a particular language. PMID- 17125750 TI - Fluoro-Jade B staining as useful tool to identify activated microglia and astrocytes in a mouse transgenic model of Alzheimer's disease. AB - Fluoro-Jade B is known as a high affinity fluorescent marker for the localization of neuronal degeneration during acute neuronal distress. However, one study suggested that fluoro-Jade B stains reactive astroglia in the primate cerebral cortex. In this study, we analyzed the staining of fluoro-Jade B alone or combined with specific markers for detection of glial fibrillary acidic protein (GFAP) or activated CD68 microglia in the double APP(SL)/PS1 KI transgenic mice of Alzheimer's disease (AD), which display a massive neuronal loss in the CA1 region of the hippocampus. Our results showed that fluoro-Jade B did not stain normal and degenerating neurons in this double mouse transgenic model. Fluoro Jade B was co-localized with Abeta in the core of amyloid deposits and in glia like cells expressing Abeta. Furthermore, fluoro-Jade B was co-localized with CD68/macrosialin, a specific marker of activated microglia, and with GFAP for astrocytes in APP(SL)/PS1 KI transgenic mice of AD. Taken together, these findings showed that fluoro-Jade B can be used to label activated microglia and astrocytes which are abundant in the brain of these AD transgenic mice. It could stain degenerating neurons as a result of acute insult while it could label activated microglia and astrocytes during a chronic neuronal degenerative process such as AD for example. PMID- 17125751 TI - Loss of calcium and increased apoptosis within the same neuron. AB - Loss of neuronal calcium is associated with later apoptotic injury but observing reduced calcium and increased apoptosis in the same cell would provide more definitive proof of this apparent correlation. Thus, following exposure to vehicle or the calcium chelator, BAPTA (1-20 microM), primary cortical neurons were labeled with Calcium Green-1 which was then cross-linked with EDAC, prior to immuno-staining for various proteins. We found that BAPTA-induced changes in calcium were highly correlated with changes in expression of activated caspase-3 as well as the calcium binding proteins calbindin, calretinin, and parvalbumin. Additionally, in brain slices from P7 neonatal rats, BAPTA induced significant loss of calcium in a brain region we have previously shown to express only moderate levels of calcium binding proteins as well as display robust apoptosis following calcium entry blockade. In contrast, BAPTA had little influence on calcium levels in a brain region we have previously shown to express robust calcium binding proteins as well as display far less apoptosis following calcium entry blockade. These data suggest that the ability of developing neurons to buffer changes in calcium may be critical to their long-term survival. PMID- 17125752 TI - Retro-convection enhanced delivery to increase blood to brain transfer of macromolecules. AB - A retro-convection enhanced delivery (R-CED) method has been developed to improve the entry of intravenously administered therapeutics within solid brain tumors. R CED uses an osmotic gradient to withdraw brain interstitial fluid (ISF) in a controlled manner via an implanted microdialysis catheter. Withdrawal of ISF increases the local tissue specific gravity in normal brain and increases twofold the extravasation of intravenous Evans blue (EB) albumin in normal brain and in an orthotopic 9L tumor. R-CED also increases the extravasation of 70 nm fluorescent liposomes fivefold in the 9L tumor. Thus the transmembrane osmotic gradient induces movement of substances in the blood into the tissue parenchyma. Following probe removal, the magnitude of the R-CED effect on EB-albumin extravasation decreases to control values within 1.5 h in normal brain; however, the effect persists beyond 6 h in the tumor. There was no evidence of histologic damage to the neurons at either 6 h or 2 weeks after R-CED. These studies establish the feasibility of applying R-CED to increase the distribution of systemically administered drugs in both the normal tissue-tumor margin as well as in the central tumor core, holding forth the possibility of improved antitumor drug efficacy. PMID- 17125753 TI - An analog of thyrotropin-releasing hormone (TRH) is neuroprotective against glutamate-induced toxicity in fetal rat hippocampal neurons in vitro. AB - TRH has been found to be efficacious in treating certain neurodegenerative disorders such as epilepsy, Alzheimer's disease, neurotrauma and depression, however, its mechanism of action is poorly understood. Since glutamate (Glu) toxicity has been implicated in these disorders, we utilized primary enriched cultures of rat fetal (E 17) hippocampal neurons to test the hypothesis that an analog of TRH, 3-Methyl-Histidine TRH (3Me-H TRH), given concurrently with Glu would protect such neurons against cell damage and cell death. Cell viability was assessed via Trypan Blue exclusion cell counts, and neuronal damage was determined by assaying lactic acid dehydrogenase (LDH) released in the conditioned media. Fetal hippocampal neurons were cultured in neurobasal media for 7 days. On day 7, neurons (10(6)/well) were treated with: control media, 10 microM 3Me-H TRH, 500 microM Glu or 500 microM Glu with either 10, 1, 0.1, 0.01 or 0.001 microM 3Me-H TRH. Both media and neurons were harvested 16 h after treatment. Prolonged exposure to 10 microM 3Me-H TRH was not toxic to the cells, whereas neurons exposed to 500 microM Glu resulted in maximal cell death. Notably, 10, 1 and 0.1 microM 3Me-H TRH, when co-treated with 500 microM Glu, protected fetal neurons against cell death in a concentration-dependent manner. These results provide support for an important neuroprotective effect of TRH/analogs against glutamate toxicity in primary hippocampal neuronal culture and implicate a potentially beneficial role of TRH/analogs in neurodegenerative diseases. PMID- 17125754 TI - Synthesis and reactivity of nonstabilized diazo sugars. AB - 1,6-Anhydro-4-deoxy-4-diazo-2,3-O-isopropylidene-beta-D-lyxo-hexopyranose (4) is a stable crystalline compound readily accessible by an improved synthetic procedure. It has been used as a model for evaluating the reactivity of the diazo group, when not stabilized by an adjacent carbonyl function, in a rigid chiral matrix. A range of carbene-type, electrophile-promoted, and 1,3-dipolar reactions were evaluated, leading to 4,4'-alkene dimers, 4-deoxy-3-enose and related derivatives, 4,4-dihalo compounds, 4-spirocyclopropane derivatives, 4 spiropyrazole structures, and by skeletal rearrangement, branched-chain anhydropentose structures having a bicyclo[2.2.2] skeleton. PMID- 17125755 TI - O-oligosaccharidyl-1-amino-1-deoxyalditols as intermediates for fluorescent labelling of oligosaccharides. AB - Reducing monosaccharides were efficiently converted to stable 1-amino-1 deoxyalditols (=glycamines; distinguished from glycosylamines by mass spectrometry) during incubation at 20 degrees C in saturated aqueous NH(4)HCO(3) containing NaCNBH(3). Potentially useful by-products included a novel, fully reduced dimer (the corresponding secondary glycamine) and several relatively long lived, unreduced products. With increasing incubation time, monomers exceeded dimers. Reducing disaccharides and oligosaccharides underwent similar reactions at their reducing termini; the yield of dimers decreased with increasing oligosaccharide M(r). The O-oligosaccharidyl-1-amino-1-deoxyalditols (OADs) obtained by reductive amination of oligosaccharides reacted readily with lissamine rhodamine sulfonyl chloride to yield OAD-sulforhodamine conjugates linked by a stable sulfonamide bond. Conditions for this reaction were optimised (borate buffer, pH9.0-9.5). The highly fluorescent OAD-sulforhodamine products were purified on a C(18) cartridge. They were electrophoretically immobile at pH2.0 and 6.5, and migrated towards the anode in borate buffer, pH9.4. The OAD sulforhodamines were amenable to TLC and were excellent substrates for enzymic transglycosylation and for glycosylhydrolase action. PMID- 17125756 TI - Preparation and characterization of N-(2-carboxybenzyl)chitosan as a potential pH sensitive hydrogel for drug delivery. AB - A novel water-soluble chitosan derivative [N-(2-carboxybenzyl)chitosan, CBCS] was synthesized. The chemical structure of CBCS was characterized by FTIR, (1)H NMR and UV spectroscopies. The degree of substitution (DS) of N-2-carboxybenzyl was determined by colloid titration. In different pH buffer solutions, the swelling characteristics of hydrogels based on CBCS (CBCSG) prepared by crosslinking with glutaraldehyde have been studied. Results showed that the swelling ratio (SR) of CBCSG decreased with an increase of the amount of glutaraldehyde, and that CBCSG swelled more significantly in alkaline solution than in acidic medium, showing the lowest SR at pH5.0. The SR of CBCSG increased with the raising of the DS of the N-2-carboxybenzyl group in alkaline solution, but no significant change was observed in an acidic environment. CBCSG showed swelling reversibility when alternately soaked in pH1.0 and 7.4 buffer solutions. Release profiles of fluorouracil (5-FU), a poorly water-soluble drug, from CBCSG were studied under both simulated gastric and intestinal pHconditions. The release was much quicker in pH7.4 buffer than in pH1.0 solution. Results indicated that CBCS could be a potential pH-sensitive carrier for colon-specific drug delivery system. PMID- 17125757 TI - Recent trends in the synthesis of O-glycosides of 2-amino-2-deoxysugars. AB - The discovery of new methods for stereoselective glycoside synthesis and convergent oligosaccharide assembly has been critical for the area of glycosciences. At the heart of this account is the discussion of the approaches for stereoselective synthesis of glycosides of 2-amino-2-deoxysugars that have emerged during the past two decades. The introductory part provides general background information and describes the key features and challenges for the synthesis of this class of compounds. Subsequently, major approaches to the synthesis of 2-amino-2-deoxyglycosides are categorized and discussed. Each subsection elaborates on the introduction (or protection) of the amino functionality, synthesis of glycosyl donors by introduction of a suitable leaving group, and glycosidation. Wherever applicable, the deprotection of a temporary amino group substituent and the conversion onto the natural acetamido functionality is described. The conclusions part evaluates the current standing in the field and provides a perspective for future developments. PMID- 17125758 TI - Dipolar relaxation in a lipid bilayer detected by a fluorescent probe, 4'' dimethylaminochalcone. AB - The dynamic behavior of polar molecules in egg phosphatidylcholine (PC) bilayers has been studied using a membrane fluorescent probe, 4''-dimethylaminochalcone (DMAC). Time and spectrally resolved fluorescence spectroscopy of DMAC incorporated in PC liposomes, as compared to studies of the probe in organic solvents, shows the existence of two independent populations, associated with different extent and speed of dipolar solvent relaxation. The first DMAC population represents approximately 69% of the fluorescence-emitting molecules, has a short fluorescence decay time (0.32 ns) and undergoes Stokes shift of 80 nm. The remaining 31% fraction of DMAC molecules has a decay time of 0.74 ns and undergoes a high (106 nm) Stokes shift. A fraction of the shift, ca. 24 nm for the first and 46 nm for the second population, is attributed to the fast (<0.1 ns) rotational relaxation of nearby dipolar molecules, which might be water. This two-state model accounts well for the detailed fluorescence properties of DMAC in egg PC, i.e. its broadened steady-state spectrum, its average fluorescence quantum yield and its complex wavelength-dependent fluorescence decays. PMID- 17125759 TI - First total syntheses of (Z)-15-methyl-10-hexadecenoic acid and the (Z)-13-methyl 8-tetradecenoic acid. AB - The first total syntheses for the (Z)-15-methyl-10-hexadecenoic acid and the (Z) 13-methyl-8-tetradecenoic acid were accomplished in seven steps and in 31-32% overall yields. The (trimethylsilyl)acetylene was the key reagent in both syntheses. It is proposed that the best synthetic strategy towards monounsaturated iso methyl-branched fatty acids with double bonds close to the omega end of the acyl chain is first acetylide coupling of (trimethylsilyl)acetylene to a long-chain bifunctional bromoalkane followed by a second acetylide coupling to a short-chain iso bromoalkane, since higher yields are thus obtained. Spectral data is also presented for the first time for these two unusual fatty acids with potential as biomarkers and as topoisomerase I inhibitors. PMID- 17125760 TI - Priming reveals differential coding of symbolic and non-symbolic quantities. AB - Number processing is characterized by the distance and the size effect, but symbolic numbers exhibit smaller effects than non-symbolic numerosities. The difference between symbolic and non-symbolic processing can either be explained by a different kind of underlying representation or by parametric differences within the same type of underlying representation. We performed a primed naming study to investigate this issue. Prime and target format were manipulated (digits or collections of dots) as well as the numerical distance between prime and target value. Qualitatively different priming patterns were observed for the two formats, showing that the underlying representations differed in kind: Digits activated mental number representations of the place coding type, while collections of dots activated number representations of the summation coding type. PMID- 17125761 TI - Analog simulation of two clinical conditions: (1) acute left ventricle failure; (2) exercise in patient with aortic stenosis. AB - A computer analysis of an equivalent electronic circuit is developed. Thus it is possible to simulate the human cardiovascular system, its negative feedback loops (including the control of venous tone, of myocardial contractility, and of heart rate) and negative intrathoracic pressure. If the simulated cardiovascular system is acted upon by various disturbances their consequences can be studied in detail. The consequences of two disturbances are studied by simulation: (i) acute left ventricular failure and (ii) exercise (decreased peripheral resistance) in aortic stenosis. However, prior to the simulation of the latter, a relatively complex condition, two additional procedures are implemented, i.e. simulations of (iii) increased sympathetic tone and of (iv) aortic stenosis are performed. Simulation of exercise (decreased peripheral resistance) in aortic stenosis is also compared with data observed in patients. Results show that, by using the present equivalent circuit, conditions described above can be qualitatively and to some extent quantitatively well simulated. PMID- 17125763 TI - Asthma, atopy, and lung function among racially diverse, poor inner-urban Minneapolis schoolchildren. AB - As part of an assessment of schoolchildren's environmental exposures and health, a probability sample of 136 children from diverse racial/ethnic backgrounds was drawn from grades 2-5 of two inner-urban Minneapolis schools (Whittier, Lyndale). Questionnaires were administered to a parent/guardian; blood samples for IgE and lung function tests were obtained. Overall adjusted rates for lifetime asthma (15.4%; 95%CI 9.3-21.5%), asthma in the last 12 months (13.6%; 7.8-19.4%), and current asthma medication use (10.5%; 5.3-15.7%) were higher than reported US national rates. Adjusted rates for lifetime physician-diagnosed asthma differed significantly among racial/ethnic groups (P<0.01): African-Americans (25.9%), White/Others (25.8%), Hispanics (9.3%), Somalis (1.8%), Asians (0%). Corresponding rates for atopy (total IgE>100 IU/mL or an allergen-specific IgE>0.35 IU/mL) were: African-Americans (66.6%), White/Others (100%), Hispanics (77.2%), Somalis (78.1%), Asians (81.8%). Lung function (FEV1, FVC) was analyzed by linear regression using log-transformed data: significant race-specific differences in lung function were found relative to White/Others (P<0.001 for each racial/ethnic group): African-Americans (FEV1 -16.5%, FVC -16.9%), Somalis ( 22.7%, -26.8%), Hispanics (-12.2%, -11.4%) and Asians (-11.1%, -12.4%). Females had significantly lower FEV1 (-8.8%) and FVC (-11.0%) than males. An unexplained, significant difference in children's lung function was found between the two schools. A history of physician-diagnosed asthma was not associated with decreased lung function. Factors other than poverty, inner-urban living, and IgE levels (atopy) need to be considered in the development of childhood asthma. PMID- 17125762 TI - Combinatorial function of ETS transcription factors in the developing vasculature. AB - Members of the ETS family of transcription factors are among the first genes expressed in the developing vasculature, but loss-of-function experiments for individual ETS factors in mice have not uncovered important early functional roles for these genes. However, multiple ETS factors are expressed in spatially and temporally overlapping patterns in the developing vasculature, suggesting possible functional overlap. We have taken a comprehensive approach to exploring the function of these factors during vascular development by employing the genetic and experimental tools available in the zebrafish to analyze four ETS family members expressed together in the zebrafish vasculature; fli1, fli1b, ets1, and etsrp. We isolated and characterized an ENU-induced mutant with defects in trunk angiogenesis and positionally cloned the defective gene from this mutant, etsrp. Using the etsrp morpholinos targeting each of the four genes, we show that the four ETS factors function combinatorially during vascular and hematopoietic development. Reduction of etsrp or any of the other genes alone results in either partial or no defects in endothelial differentiation, while combined reduction in the function of all four genes causes dramatic loss of endothelial cells. Our results demonstrate that combinatorial ETS factor function is essential for early endothelial specification and differentiation. PMID- 17125764 TI - Inhibitory effect of CDP, a polysaccharide extracted from the mushroom Collybia dryophila, on nitric oxide synthase expression and nitric oxide production in macrophages. AB - The effect of Collybia dryophila polysaccharide (CDP), a (1-->3), (1-->4)-beta-D glucan extracted from the mushroom C. dryophila, was evaluated on nitric oxide (NO) production induced by lipopolysaccharide (LPS) and gamma interferon (IFNgamma) or by LPS alone in RAW 264.7 cells. CDP significantly inhibited NO production in a dose-dependent manner without affecting cell viability. The inhibition of NO by CDP was consistent with decreases in both inducible nitric oxide synthase (iNOS) protein and mRNA expression suggesting that CDP exerts its effect by inhibiting iNOS gene expression. In addition, CDP at concentrations of 400 and 800 microg/ml was shown to significantly increase prostaglandin E2 (PGE2) production in LPS- and IFNgamma-induced macrophages when compared to the control. PMID- 17125765 TI - Prevention of pro-depressant effect of L-arginine in the forced swim test by NG nitro-L-arginine and [1H-[1,2,4]Oxadiazole[4,3-a]quinoxalin-1-one]. AB - Previous studies have shown that l-arginine, the precursor of nitric oxide, has a dual effect (antidepressant and pro-depressant) in the forced swim test. The aim of the present study was to investigate whether nitric oxide-cGMP pathway was involved in this dual effect. Porsolt swim test was conducted to resemble the symptomatology of major depressive disorder. An open field locomotor activity test was also used. L-arginine exerted a U-shape effect in the forced swim test: doses of 30, 100, 300, and 1000 mg/kg caused no alteration, statistically significant reduction, no alteration, and non-significant enhancement, respectively. Neither N(G)-nitro-L-arginine (NNA) nor [1H-[1,2,4]Oxadiazole[4,3 a]quinoxalin-1-one] (ODQ) at doses of 3 mg/kg was found to be effective in the forced swim test, whereas 10 mg/kg ODQ significantly reduced the immobility time. In the presence of NNA, the antidepressant and pro-depressant effects of L arginine disappeared, however, only the pro-depressant component of l-arginine effect was prevented by ODQ (3 and 10 mg/kg). Saline, the solvent of L-arginine and NNA, and dimethyl sulfoxide (15% in saline), the solvent of ODQ, had no effect on the duration of immobility. None of the drugs or solvents used in the present study had any effect on locomotor activity over the dose range applied. The results show that L-arginine exerts its paradoxical effects by producing nitric oxide and that cGMP seems to have a role only in the pro-depressant component. PMID- 17125766 TI - Hip-phase-dependent flexion reflex modulation and expression of spasms in patients with spinal cord injury. AB - The flexion reflex in human spinal cord injury (SCI) is believed to incorporate interneuronal circuits that consist elements of the stepping generator while ample evidence suggest that hip proprioceptive input is a controlling signal of locomotor output. In this study, we examined the expression of the non nociceptive flexion reflex in response to imposed sinusoidal passive movements of the ipsilateral hip in human SCI. The flexion reflex was elicited by low intensity stimulation (300 Hz, 30 ms pulse train) of the right sural nerve at the lateral malleolus, and recorded from the tibialis anterior (TA) muscle. Sinusoidal hip movements were imposed to the right hip joint at 0.2 Hz by a Biodex system while subjects were supine. The effects of leg movement on five leg muscles along with hip, knee, and ankle joint torques were established simultaneously with the modulation pattern of the flexion reflex during hip oscillations. Phase-dependent modulation of the flexion reflex was present during hip movement, with the reflex to be significantly facilitated during hip extension and suppressed during hip flexion. The phase-dependent flexion reflex modulation coincided with no changes in TA pre- and post-stimulus background ongoing activity during hip extension and flexion. Reflexive muscle and joint torque responses, induced by the hip movement and substantiated by excitation of flexion reflex afferents, were entrained to specific phases of hip movement. Joint torque responses were consistent with multi-joint spasmodic muscle activity, which was present mostly during the transition phase of the hip from flexion to extension and from mid- to peak extension. Our findings provide further evidence on the interaction of hip proprioceptors with spinal interneuronal circuits engaged in locomotor pathways, and such interaction should be considered in rehabilitation protocols employed to restore sensorimotor function in people with SCI. PMID- 17125768 TI - Mild surfection of neural cells, especially motoneurons, in primary culture and cell lines. AB - Of all cell types, motoneurons (MNs), are possibly the most difficult to maintain in culture, since their development and survival is conditioned by many factors that are still in the course of identification. This may also be the reason why they are difficult to transfect. We succeed to transfect these fragile cells with lipoplex [DOTAP:PC (10:1)-pGFP]-precoated coverslips. Here, we report that this original method, also termed 'surfection' does not perturbate MN development and survival while giving important transfection yield (15%). Lipofectamine 2000 and other well-known auxiliary lipids (DOPE, Chol) give lower surfection yields. The use of (DOTAP:PC)-based lipid vector also can be extended to several neural and non-neural cell lines with appreciable transfection yield such as a glial cell line (GCL) derived from rat spinal cord (65%), HeLa S3 (60%), COS-7 (30%) and HEK 293 cells (20%). The efficiency of DOTAP:PC (10:1) and Lipofectamine 2000 vectors in our surfection method are compared on standard HeLa S3 cell lines. Lipofectamine 2000 (72%) is slightly better than DOTAP:PC (10:1) (60%). However, the surfection method improved the efficiency of Lipofectamine 2000 itself (72%) as compared to the classical (62%) approach. In summary we have developed an original standard surfection protocol for both MN primary cultures and cell lines, thus simplifying laboratory practice; moreover, Lipofectamine 2000 used in this surfection method is more efficient for the cell lines than the manufacturer recommended method. We emphasize that our method particularly spares fragile cells like MNs from injure and therefore, might be applied to other fragile cell type in primary cultures. PMID- 17125767 TI - Evidence of GLP-1-mediated neuroprotection in an animal model of pyridoxine induced peripheral sensory neuropathy. AB - Pyridoxine (vitamin B6) intoxicated rodents develop a peripheral neuropathy characterized by sensory nerve conduction deficits associated with disturbances of nerve fiber geometry and axonal atrophy. To investigate the possibility that glucagon-like peptide-1 (7-36)-amide (GLP-1) receptor agonism may influence axonal structure and function through neuroprotection neurotrophic support, effects of GLP-1 and its long acting analog, Exendin-4 (Ex4) treatment on pyridoxine-induced peripheral neuropathy were examined in rats using behavioral and morphometric techniques. GLP-1 is an endogenous insulinotropic peptide secreted from the gut in response to the presence of food. GLP-1 receptors (GLP 1R) are coupled to the cAMP second messenger pathway, and are expressed widely throughout neural tissues of humans and rodents. Recent studies have established that GLP-1 and Ex4, have multiple synergistic effects on glucose-dependent insulin secretion pathways of pancreatic beta-cells and on neural plasticity. Data reported here suggest that clinically relevant doses of GLP-1 and Ex4 may offer some protection against the sensory peripheral neuropathy induced by pyridoxine. Our findings suggest a potential role for these peptides in the treatment of neuropathies, including that associated with type II diabetes mellitus. PMID- 17125769 TI - Constitutive activation of the ERK-MAPK pathway in the suprachiasmatic nuclei inhibits circadian resetting. AB - Circadian entrainment involves photic stimulation of the suprachiasmatic molecular oscillator, including activation of the ERK/MAP kinase, which is phosphorylated endogenously during the day and in response to light during the night. We aimed to disrupt the diurnal cycle of ERK phosphorylation by in vivo transfection of a constitutively active form of MEK, a MAPK kinase. This procedure did not affect normal circadian parameters, but completely inhibited light-induced phase advances. Therefore, circadian regulation of the ERK pathway is not essential for the normal mechanism of the biological clock, but it is fundamental as an interface with environmental entrainment by light. PMID- 17125770 TI - Inhibition of the formation of amyloid beta-protein fibrils using biocompatible nanogels as artificial chaperones. AB - The formation of fibrils by amyloid beta-protein (Abeta) is considered as a key step in the pathology of Alzheimer's disease (AD). Inhibiting the aggregation of Abeta is a promising approach for AD therapy. In this study, we used biocompatible nanogels composed of a polysaccharide pullulan backbone with hydrophobic cholesterol moieties (cholesterol-bearing pullulan, CHP) as artificial chaperones to inhibit the formation of Abeta-(1-42) fibrils with marked amyloidgenic activity and cytotoxicity. The CHP-nanogels incorporated up to 6-8 Abeta-(1-42) molecules per particle and induced a change in the conformation of Abeta from a random coil to alpha-helix- or beta-sheet-rich structure. This structure was stable even after a 24-h incubation at 37 degrees C and the aggregation of Abeta-(1-42) was suppressed. Furthermore, the dissociation of the nanogels caused by the addition of methyl-beta-cyclodextrin released monomeric Abeta molecules. Nanogels composed of amino-group-modified CHP (CHPNH(2)) with positive charges under physiological conditions had a greater inhibitory effect than CHP-nanogels, suggesting the importance of electrostatic interactions between CHPNH(2) and Abeta for inhibiting the formation of fibrils. In addition, CHPNH(2) nanogels protected PC12 cells from Abeta toxicity. PMID- 17125771 TI - Suppressive effects of diacylglycerol oil on postprandial hyperlipidemia in insulin resistance and glucose intolerance. AB - The diacylglycerol (DAG), a commonly used as a cooking oil in Japan, results in a lower elevation of serum triglyceride (TG) after ingestion compared to triacylglycerol (TAG). Postprandial hyperlipidemia (PPHL) and an increase in remnant lipoproteins (RLP) levels are risk factors for CAD, and a close relationship between PPHL and type 2 diabetes and/or insulin resistance has been reported. To evaluate the effect of DAG on PPHL in insulin resistance and glucose intolerance, 11 subjects with a normal glucose tolerance (NGT) and 14 subjects with IGT received oral fat tolerance test (OFTT) twice. They ingested emulsified test oils prepared with either DAG or TAG. In the IGT subjects, after the DAG and TAG load, the serum concentrations of TG, RLP-TG, and RLP-cholesterol increased throughout the 4-h study. The responses of these variables above baseline after the DAG load were significantly smaller than those after the TAG load (p<0.05). In contrast, in the NGT subjects, changes in these parameters were much smaller than those observed for IGT subjects. The difference in the integrated responses for serum RLP-cholesterol concentration during OFTT between DAG and TAG in all subjects can be easily explained by the integrated response of insulin rather than glucose during oral glucose tolerance test (r=0.7, p<0.01). DAG was more effective in insulin resistant and hyperinsulinemic participants regardless of glucose intolerance, and may be beneficial in reducing the extent of CAD risk in such individuals. PMID- 17125772 TI - Response to 'Laurberg JM, Olsen AK, Hansen SB, et al. Imaging of vulnerable atherosclerotic plaques with FDG-microPET: no FDG accumulation' [Atherosclerosis 2006]. PMID- 17125773 TI - The relationship between endothelial function in the brachial artery and intima plus media thickening of the coronary arteries in patients with chest pain syndrome. AB - Endothelial dysfunction precedes the development of clinical atherosclerosis. A decrease in endothelium-derived nitric oxide activity shows impaired vasodilator function and causes arterial intimal hyperplasia or thickening. Others and we have reported the close relation of endothelial function in brachial artery and coronary artery. To compare the flow-mediated endothelium-dependent vasodilation (FMD) in brachial artery and the intima+media area in coronary artery, we measured brachial artery vasodilator response following transient occlusion with high resolution ultrasound transducer and intima+media area in coronary arteries with intravascular ultrasound in 25 patients with normal coronary angiograms (age 61.6+/-8.7 years old, men 14 and women 11). FMD was measured at least 7 days after the cessation of all vasodilators. The mean FMD of 25 patients was 3.83+/ 2.38%, the mean intima+media area in coronary arteries of 25 patients was 39.9+/ 15.5% of total vessel wall. FMD has a close negative relation with the largest percent intima+media area (r=-0.77, p<0.01). Especially, the patients whose FMD was less than 3.83% had larger percent intima+media area than those whose FMD was 3.83% or more (48.7+/-10.7% versus 30.3+/-14.2%, p<0.01). There is an intima+media thickening even in the patients who had normal coronary angiograms, and that the percent intima+media area correlated with FMD. The measurement of FMD is useful for screening the coronary artery intima+media thickening noninvasively. The pathogenesis of acute coronary syndrome has been reported to be the plaque rupture even in the patients with normal coronary angiograms. Thus, we must pay much attention in the patients with impaired FMD even in the normal coronary angiograms. PMID- 17125774 TI - Serum paraoxonase-1 activity in Helicobacter pylori infected subjects. AB - Previous studies have suggested that Helicobacter pylori (H. pylori) infection may play an important role in the process of atherosclerosis. The objective of this study was to investigate serum paraoxonase and arylesterase activities, and lipid hydroperoxide (LOOH) and total thiol (SH) levels along with lipid parameters in H. pylori infected subjects. Fifty-six H. pylori positive subjects and 43 H. pylori negative subjects were enrolled. Serum paraoxonase and arylesterase activities were measured spectrophotometrically. LOOH levels were measured by FOX-2 assay. Serum SH levels, paraoxonase and arylesterase activities were significantly lower in H. pylori positive group than H. pylori negative group (all p<0.05), while LOOH levels were significantly higher (p<0.05). In H. pylori positive subjects, serum LOOH levels were correlated with SH levels (r= 0.247, p<0.05), serum paraoxonase (r=-0.432, p<0.05) and arylesterase activities (r=-0.404, p<0.001), and triglyceride (r=0.305, p<0.05), total cholesterol (r=0.568, p<0.05), high-density lipoprotein-cholesterol (HDL-C) (r=-0.300, p<0.05) and low-density lipoprotein-cholesterol (LDL-C) (r=0.577, p<0.05) levels. Serum paraoxonase and arylesterase activities were also correlated with HDL-C levels (r=0.221, p<0.05 and r=0.291, p<0.05, respectively), while no correlation was observed with triglyceride, total cholesterol and LDL-C levels (both p>0.05). In conclusion, paraoxonase and arylesterase activities decrease significantly in H. pylori infected subjects. Lower serum paraoxonase-1 (PON1) activity seems to be related to decrease in HDL-C and, in part, to increased oxidative stress and inflammatory condition induced by H. pylori infection. Measurement of serum PON1 activity may help in the early identification of H. pylori infected subjects with increased risk of atherosclerotic disease. PMID- 17125775 TI - Mechanical properties of soft human tissues under dynamic loading. AB - The dynamic response of soft human tissues in hydrostatic compression and simple shear is studied using the Kolsky bar technique. We have made modifications to the technique that allow loading of a soft tissue specimen in hydrostatic compression or simple shear. The dynamic response of human tissues (from stomach, heart, liver, and lung of cadavers) is obtained, and analyzed to provide measures of dynamic bulk modulus and shear response for each tissue type. The dynamic bulk response of these tissues is easily described by a linear fit for the bulk modulus in this pressure range, whereas the dynamic shearing response of these tissues is strongly non-linear, showing a near exponential growth of the shear stress. PMID- 17125776 TI - Three-dimensional inhomogeneous triphasic finite-element analysis of physical signals and solute transport in human intervertebral disc under axial compression. AB - A 3D inhomogeneous finite-element model for charged hydrated soft tissues containing charged/uncharged solutes was developed and applied to analyze the mechanical, chemical, and electrical signals within the human intervertebral disc during an axial unconfined compression. The effects of tissue properties and boundary conditions on the physical signals and the transport of fluid and solute were investigated. The numerical simulation showed that, during disc compression, the fluid pressurization and the effective (von Misses) solid stress were more pronounced in the annulus fibrosus (AF) region near the interface between AF and nucleus pulposus (NP). In NP, the distributions of the fluid pressure, effective stress, and electrical potential were more uniform than those in AF. The electrical signals were very sensitive to fixed charge density. Changes in material properties of NP (water content, fixed charge density, and modulus) affected fluid pressure, electrical potential, effective stress, and solute transport in the disc. This study is important for understanding disc biomechanics, disc nutrition, and disc mechanobiology. PMID- 17125777 TI - Development of a headspace-solid phase micro extraction method to monitor changes in volatile profile of rose (Rosa hybrida, cv David Austin) petals during processing. AB - In the present study, headspace solid phase microextraction combined to capillary gas chromatography (HS-SPME-GC) has been applied for the determination of changes in the volatile profile of rose petals (Rosa hybrida, cvs David Austin) following processing (heat treatment and addition as an ingredient to a food product--for example yoghurt). Four SPME fibres at two sampling temperatures (40 and 60 degrees C) with a sampling time of 30 min were examined. Volatile profiles were detected either by FID or/and by olfactometry (ODP-II, Gerstel). Fibre testing was performed using raw rose petals for sampling temperature selection and an 18 characteristic rose volatile standard mixture in water was used to compare fibre performances at the sampling temperature of 60 degrees C. Polydimethylsiloxane divinylbenzene (PDMS-DVB) fibre at the sampling temperature of 60 degrees C was the most suitable to sample the rose alcohols phenyl ethanol, citronellol, nerol, geraniol and eugenol, as assessed by GC-olfactometry, not only from raw petals, but also from processed rose petals and the food product. PDMS-DVB fibre also showed a desired low affinity to volatiles from yoghurt, which reduces the influence of food matrix on the volatile profile. The method was linear over two orders of magnitude and had satisfactory repeatability, with limits of detection for the rose alcohols ranging from <1 to 10 ng/ml concentration levels. PMID- 17125778 TI - High-throughput quantification of lincomycin traces in fermentation broth of genetically modified Streptomyces spp. Comparison of ultra-performance liquid chromatography and high-performance liquid chromatography with UV detection. AB - A new separation and quantification method using ultra-performance liquid chromatography (UPLC) with UV detection was developed for detection of lincomycin traces in fermentation broth of different Streptomyces spp. A similar high performance liquid chromatography (HPLC) protocol was simultaneously developed for comparison purposes. Both methods were validated and showed a linear range of detector response for quantification of lincomycin in concentration from 3.125 to 1000.0 microgml(-1) with correlation coefficient 0.999 and recoveries ranging from 81.5 to 89.85% with precision < or =5%. Compared with the HPLC, the UPLC method offered high sample throughput and about 10 times lower consumption of solvents. The developed assays were used for determination of lincomycin production in genetically manipulated production strain Streptomyces lincolnensis and for determination of lincomycin production after heterologous expression of lincomycin biosynthetic gene cluster in non-producing strain Streptomyces coelicolor. PMID- 17125779 TI - Liquid chromatography/atmospheric pressure ionization mass spectrometry with post column liquid mixing for the efficient determination of partially oxidized polycyclic aromatic hydrocarbons. AB - The analytical hyphenation of micro-flow high-performance liquid chromatography (LC), with post-column liquid mixing and mass spectrometric detection (MS) was established to detect partially oxidized polycyclic aromatic hydrocarbons (oxy PAHs) for low quantity samples. 100pmol injections of 30 reference standards could be detected in good sensitivity using either atmospheric pressure chemical ionization (APCI) and/or atmospheric pressure photoionization (APPI). The connected mass spectrometer was a single quadrupol analyzer realizing simultaneous registration of positive and negative ions in scan range width of 200 - 300Da. The ionization efficiency was compared using three ionization sources (incl. electrospray ionization (ESI)) for several oxy-PAHs. According to the mass spectra, the analytes behave differently in ionization properties. Ionization mechanism (e.g. deprotonated ions and electron captured ions) could be discussed with new inside views. Finally, the hyphenated system was applied to an exemplary aerosol extract and thus highlighting the expedient utilization of this downscaled method for real samples. PMID- 17125780 TI - Assay of glutathione in must and wines using capillary electrophoresis and laser induced fluorescence detection. Changes in concentration in dry white wines during alcoholic fermentation and aging. AB - Glutathione (GSH) was assayed in must and wine using capillary electrophoresis coupled with laser-induced fluorescence (LIF) detection. Sample preparation involved conjugating thiols with monobromobimane (MBB) in a 2-(N cyclohexylamino)ethanesulfonic acid [CHES] buffer (179mM). The electrophoretic conditions were 30kV with a capillary length of 105cm from the inlet to the detector (120cm total length) and a 50microm inner diameter. Under these conditions, the complete separation from the other main non-volatile thiols took less than 20min. We also described the optimum conditions for derivatizing wine samples with MBB to increase eletrophoretic sensitivity. The detection limit for glutathione assay is 65nmol/L. This simple, sensitive method provides a specific assay of glutathione in reduced form, as the sample preparation technique does not modify the balance of oxidized and reduced forms. We used this method to monitor changes in the reduced glutathione content of a white wine during alcoholic fermentation and barrel aging. PMID- 17125781 TI - How many phases and phase transitions do exist in Gibbs adsorption layers at the air-water interface? AB - Four different phases and four different first-order phase transitions have been shown to exist in Gibbs adsorption layers of mixtures containing n-hexadecyl dihydrogen phosphate (n-HDP) and L-arginine (L-arg) at a molar ratio of 1:2. These conclusions have been made from surface pressure-time (pi-t) adsorption isotherms measured with a film balance and from monolayer morphology observed with a Brewster angle microscopy (BAM). The observed four phases are gas (G), liquid expanded (LE), liquid condensed (LC) and LC' phases. Three first-order phase transitions are G-LE, LE-LC and LC-LC'. However, the thermodynamically allowed G-LC phase transition in a 1.2 x 10(-4) M mixture at 2 degrees C, which is below the so-called triple point, is kinetically separated into the G-LE and LE-LC phase transitions. The most interesting observation is that the homogeneous LC phase shows a new first-order phase transition named as LC-LC' at 2 or 5 degrees C. The LE and LC phases represent circular and fractal shaped domains, respectively, whereas the LC' phase shows very bright, anisotropic and characteristic shaped domains. PMID- 17125782 TI - Monte Carlo modeling of ion adsorption at the energetically heterogeneous metal oxide/electrolyte interface: Micro- and macroscopic correlations between adsorption energies. AB - Grand canonical Monte Carlo simulations are carried out for the basic Stern model of the electrical double layer formed at the energetically heterogeneous metal oxide/electrolyte interface. The effect of the global (macroscopic) and local (microscopic) adsorption energies correlations as well as the influence of the model parameter on the surface charge density curves were investigated. The linear dependence of point of zero charge (PZC) as a function of H+ ion adsorption energy proves that the acidic/basic properties of the system are mainly governed by proton uptake/release. Two kinds of systems were taken into account: one neglecting lateral interactions and the other one including them. The effect of electrolyte concentrations as well as the surface heterogeneity on the surface charge density curves were shown too. The presented simulation algorithm allows to model two experimentally observed instances of the metal oxide/electrolyte interface: one possessing a common intersection point (CIP) at pH = PZC and the other one with CIP not equal PZC. PMID- 17125783 TI - A study on pore-opening behaviors of graphite nanofibers by a chemical activation process. AB - In this work, porous graphite nanofibers (GNFs) were prepared by a KOH activation method in order to manufacture porous carbon nanofibers. The process was conducted in the activation temperature range of 900-1100 degrees C, and the KOH:GNFs ratio was fixed at 3.5:1. The textural properties of the porous carbons were analyzed using N2 adsorption isotherms at 77 K. The BET, D-R, and BJH equations were used to observe the specific surface areas and the micro- and mesopore structures, respectively. From the results, it was found that the textural properties, including the specific surface area and the pore volumes, were proportionally enhanced with increasing activation temperatures. However, the activation mechanisms showed quite significant differences between the samples activated at low and high temperatures. PMID- 17125784 TI - Dispersion stability of TiO2 nanoparticles covered with SiOx monolayers in water. AB - The repetition of a two-step route consisting of chemisorption of 1,3,5,7 tetramethylcyclotetrasiloxane (TMCTS) and its subsequent photooxidation has formed SiOx monolayers on the surfaces of TiO2 particles layer by layer (SiOx(n < or = 4)/TiO2, n = repeated number). The trace of these processes using diffuse reflectance FT-IR and solid-state 29Si NMR spectroscopy reveals that TMCTS is chemisorbed on TiO2 through Ti-O-Si bonds followed by growth of Si-O-Si networks via dehydration-condensation of the Si-OH groups generated by TiO2 photocatalytic oxidation of the Si-H and Si-CH3 groups. The point of zero charge of the TiO2 particles decreases from 6.5 to 5.4 at n = 1, reaching 4.9 at n2. As a result of coverage with the SiOx monolayers, the dispersion stability of TiO2 particles in neutral water is significantly improved without changing their optical properties. The dispersion stability further increases with accumulation of SiOx monolayers. PMID- 17125785 TI - Satisfaction with communicative participation as defined by adults with multiple sclerosis: a qualitative study. AB - PURPOSE: This study examined satisfaction with communicative participation as reported by adults with multiple sclerosis (MS). METHOD: Eight community-dwelling adults with MS participated in semi-structured interviews. They were asked to discuss their satisfaction with their communication in a variety of situations. Interviews were analyzed using a constant comparative method of qualitative description. RESULTS: Themes derived included: Comfort, consisting of Ease and Confidence; Success of the Outcome, including Function is Achieved and A Connection is Made; and Personal Meaning of Participation, including Personal Preferences, Comparison with the Past, and Thinking about One's Own Communication. CONCLUSIONS: Participants described multiple facets of satisfaction with communicative participation. Some of the dimensions were similar to those in existing assessment instruments such as levels of ease or difficulty with performance. Participants did not talk about frequency of activities as a key part of their satisfaction. Implications for identifying intervention targets and treatment outcome measurements are provided. LEARNING OUTCOMES: The reader should be able to: 1) define communicative participation and identify key elements of this construct; 2) identify the issues that were most relevant to satisfaction with communicative participation with participants with MS; and 3) identify reasons for greater emphasis on the subjective viewpoint of people with communication disorders in measurement of treatment outcomes. PMID- 17125789 TI - Infection of Ixodes scapularis ticks with Rickettsia monacensis expressing green fluorescent protein: a model system. AB - Ticks (Acari: Ixodidae) are ubiquitous hosts of rickettsiae (Rickettsiaceae: Rickettsia), obligate intracellular bacteria that occur as a continuum from nonpathogenic arthropod endosymbionts to virulent pathogens of both arthropod vectors and vertebrates. Visualization of rickettsiae in hosts has traditionally been limited to techniques utilizing fixed tissues. We report epifluorescence microscopy observations of unfixed tick tissues infected with a spotted fever group endosymbiont, Rickettsia monacensis, transformed to express green fluorescent protein (GFP). Fluorescent rickettsiae were readily visualized in tick tissues. In adult female, but not male, Ixodes scapularis infected by capillary feeding, R. monacensis disseminated from the gut and infected the salivary glands that are crucial to the role of ticks as vectors. The rickettsiae infected the respiratory tracheal system, a potential dissemination pathway and possible infection reservoir during tick molting. R. monacensis disseminated from the gut of capillary fed I. scapularis nymphs and was transstadially transmitted to adults. Larvae, infected by immersion, transstadially transmitted the rickettsiae to nymphs. Infected female I. scapularis did not transovarially transmit R. monacensis to progeny and the rickettsiae were not horizontally transmitted to a rabbit or hamsters. Survival of infected nymphal and adult I. scapularis did not differ from that of uninfected control ticks. R. monacensis did not disseminate from the gut of capillary fed adult female Amblyomma americanum (L.), or adult Dermacentor variabilis (Say) ticks of either sex. Infection of I. scapularis with R. monacensis expressing GFP provides a model system allowing visualization and study of live rickettsiae in unfixed tissues of an arthropod host. PMID- 17125790 TI - Host mediated selection of pathogen genotypes as a mechanism for the maintenance of baculovirus diversity in the field. AB - The genetic diversity of many DNA virus populations in nature is unknown, but for those that have been studied it has been found to be relatively high. This is particularly true for baculoviruses, a family of large double-stranded DNA viruses that infect the larval stages of insects. Why there should be such heterogeneity within these virus populations is puzzling and what sustains it is still unknown. It has long been recognized that some baculoviruses have a relatively wide host range, but the effect of different host species on the genotypic structure of a baculovirus population has received little attention. We provide evidence that infection of different insect species can influence the genetic diversity of a Panolis flammea nucleopolyhedrovirus (PaflNPV) population, isolated from the pine beauty moth. Variable regions of the PaflNPV genome were sequenced and novel ORFs were identified on each of the enlarged fragments. The roles of these orfs and the implications of their presence or absence within different genotypes are discussed. The variable fragments were also labelled with 32P and used as polymorphic genetic markers of genotype abundance. The proportion of polymorphic loci changed after passage in different insect species and this varied among species, suggesting a role for host selection of pathogen genotypes in the field as a mechanism for maintaining genetic diversity. These results have wide-ranging implications for understanding the ecology of insect-virus interactions in the natural environment and the evolution of baculovirus life history strategies. PMID- 17125791 TI - Probing Interactions within the synaptic DNA-SfiI complex by AFM force spectroscopy. AB - SfiI belongs to a family of restriction enzymes that function as tetramers, binding two recognition regions for the DNA cleavage reaction. The SfiI protein is an attractive and convenient model for studying synaptic complexes between DNA and proteins capable of site-specific binding. The enzymatic action of SfiI has been very well characterized. However, the properties of the complex before the cleavage reaction are not clear. We used single-molecule force spectroscopy to analyze the strength of interactions within the SfiI-DNA complex. In these experiments, the stability of the synaptic complex formed by the enzyme and two DNA duplexes was probed in a series of approach-retraction cycles. In order to do this, one duplex was tethered to the surface and the other was tethered to the probe. The complex was formed by the protein present in the solution. An alternative setup, in which the protein was anchored to the surface, allowed us to probe the stability of the complex formed with only one duplex in the approach retraction experiments, with the duplex immobilized at the probe tip. Both types of complexes are characterized by similar rupture forces. The stability of the complex was determined by measuring the dependence of rupture forces on force loading rates (dynamic force spectroscopy) and the results suggest that the dissociation reaction of the SfiI-DNA complex has a single energy barrier along the dissociation path. Dynamic force spectroscopy was instrumental in revealing the role of the 5 bp spacer region within the palindromic recognition site on DNA SfiI in the stability of the complex. The data show that, although the change of non-specific sequence does not alter the position of the activation barrier, it changes values of the off rates significantly. PMID- 17125792 TI - Action of transcription factors in the control of transferrin receptor expression in human brain endothelium. AB - Brain endothelium has a distinctive phenotype, including high expression of transferrin receptor, p-glycoprotein, claudin-5 and occludin. Dermal endothelium expresses lower levels of the transferrin receptor and it is absent from lung endothelium. All three endothelia were screened for transcription factors that bind the transferrin receptor promoter and show different patterns of binding between the endothelia. The transcription factor YY1 has distinct DNA-binding activities in brain endothelium and non-brain endothelium. The target-sites on the transferrin receptor promotor for YY1 lie in close proximity to those of the transcription initiation complex containing TFIID, so the two transcription factors potentially compete or interfere. Notably, the DNA-binding activity of TFIID was the converse of YY1, in different endothelia. YY1 knockdown reduced transferrin receptor expression in brain endothelium, but not in dermal endothelium, implying that YY1 is involved in tissue-specific regulation of the transferrin receptor. Moreover a distinct YY1 variant is present in brain endothelium and it associates with Sp3. A model is presented, in which expression from the transferrin receptor gene in endothelium requires the activity of both TFIID and Sp3, but whether the gene is transcribed in different endothelia, is related to the balance between activating and suppressive forms of YY1. PMID- 17125793 TI - Structural basis of enhanced binding of extended and helically constrained peptide epitopes of the broadly neutralizing HIV-1 antibody 4E10. AB - Potent, broadly HIV-1 neutralizing antibodies (nAbs) may be invaluable for the design of an AIDS vaccine. 4E10 is the broadest HIV-1 nAb known to date and recognizes a contiguous and highly conserved helical epitope in the membrane proximal region of gp41. The 4E10 epitope is thus an excellent target for vaccine design as it is also highly amenable to peptide engineering to enhance its helical character. To investigate the structural effect of both increasing the peptide length and of introducing helix-promoting constraints in the 4E10 epitope, we have determined crystal structures of Fab 4E10 bound to an optimized peptide epitope (NWFDITNWLWYIKKKK-NH(2)), an Aib-constrained peptide epitope (NWFDITNAibLWRR-NH(2)), and a thioether-linked peptide (NWFCITOWLWKKKK-NH(2)) to resolutions of 1.7 A, 2.1 A, and 2.2 A, respectively. The thioether-linked peptide is the first reported structure of a cyclic tethered helical peptide bound to an antibody. The introduced helix constraints limit the conformational flexibility of the peptides without affecting interactions with 4E10. The substantial increase in affinity (10 nM versus 10(4) nM of the IC(50) of the original KGND peptide template) is largely realized by 4E10 interaction with an additional helical turn at the peptide C terminus that includes Leu679 and Trp680. Thus, the core 4E10 epitope was extended and modified to a WFX(I/L)(T/S)XX(L/I)W motif, where X does not play a major role in 4E10 binding and can be used to introduce helical-promoting constraints in the peptide epitope. PMID- 17125794 TI - The application of the psychological contract to workplace safety. AB - INTRODUCTION: Psychological contracts of safety are conceptualized as the beliefs of individuals about reciprocal safety obligations inferred from implicit or explicit promises. Although the literature on psychological contracts is growing, the existence of psychological contracts in relation to safety has not been established. The research sought to identify psychological contracts in the conversations of employees about safety, by demonstrating reciprocity in relation to employer and employee safety obligations. The identified safety obligations were used to develop a measure of psychological contracts of safety. METHOD: The participants were 131 employees attending safety training sessions in retail and manufacturing organizations. Non-participant observation was used to collect the data during safety training sessions. Content analysis was used to analyze the data. Categories for coding were established through identification of language markers that demonstrated contingencies or other implied obligations. RESULTS: Direct evidence of reciprocity between employer safety obligations and employee safety obligations was found in statements from the participants demonstrating psychological contracts. A comprehensive list of perceived employer and employee safety obligations was compiled and developed into a measure of psychological contracts of safety. A small sample of 33 safety personnel was used to validate the safety obligations. CONCLUSIONS AND IMPACT ON INDUSTRY: Implications of these findings for safety and psychological contract research are discussed. PMID- 17125795 TI - Spontaneous recanalization at subacute phase of stroke may be dramatic: A case report. AB - Spontaneous recanalization of the occluded cerebral arteries has occasionally occurred in an acute phase of stroke patients. We report a stroke patient with recanalization at 7 days of onset detected by using continuous transcranial Doppler (TCD) monitoring. After the presence of microembolic signals on TCD display, dynamic flow changes of the middle cerebral artery (MCA) occurred, which represented recanalization of the occluded MCA. PMID- 17125796 TI - A novel compound heterozygous mutation in the DAP12 gene in a patient with Nasu Hakola disease. AB - A 34-year-old woman showed clinical features characteristic of Nasu-Hakola disease (NHD), also designated polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy (PLOSL). The genetic analysis of the DAP12 gene (TYROBP) identified two heterozygous mutations composed of a previously reported single base deletion of 141G (141delG) in exon 3 and a novel single base substitution of G262T in exon 4, both of which are located on separate alleles. The protein sequence motif search indicated that both mutations encode truncated nonfunctional DAP12 polypeptides. This is the first case of NHD caused by compound heterozygosity for loss-of-function mutations in DAP12. PMID- 17125797 TI - Interferon beta-1b exacerbates multiple sclerosis with severe optic nerve and spinal cord demyelination. AB - To evaluate the effect of interferon beta-1b (IFNB-1b) on multiple sclerosis (MS) with severe optic nerve and spinal cord demyelination, we examined the relationship between IFNB-1b treatment outcome and the clinical and genetic characteristics of three types of demyelinating diseases of the central nervous system, i.e., neuromyelitis optica (NMO), MS and MS with severe optic-spinal demyelination. Japanese MS frequently carried HLA DPB1*0501, which is associated with NMO. MS with DPB1*0501 showed severe optic-spinal demyelination represented by longitudinally extensive spinal cord lesion, blindness and CSF pleocytosis. IFNB-1b treatment did not succeed in these patients because of the increase of optic nerve and spinal cord relapse and other severe side effects. IFNB-1b should not be administered to demyelinating patients with genetic and clinical characteristics mimicking NMO such as HLA DPB1*0501 allele, longitudinally extensive spinal cord lesion, blindness and CSF pleocytosis even if they have symptomatic cerebral lesions as typically seen in MS. The present study strongly suggests that these patients should be diagnosed as having NMO. PMID- 17125798 TI - Evolutionary dynamics of the continuous iterated prisoner's dilemma. AB - The iterated prisoner's dilemma (IPD) has been widely used in the biological and social sciences to model dyadic cooperation. While most of this work has focused on the discrete prisoner's dilemma, in which actors choose between cooperation and defection, there has been some analysis of the continuous IPD, in which actors can choose any level of cooperation from zero to one. Here, we analyse a model of the continuous IPD with a limited strategy set, and show that a generous strategy achieves the maximum possible payoff against its own type. While this strategy is stable in a neighborhood of the equilibrium point, the equilibrium point itself is always vulnerable to invasion by uncooperative strategies, and hence subject to eventual destabilization. The presence of noise or errors has no effect on this result. Instead, generosity is favored because of its role in increasing contributions to the most efficient level, rather than in counteracting the corrosiveness of noise. Computer simulation using a single locus infinite alleles Gaussian mutation model suggest that outcomes ranging from a stable cooperative polymorphism to complete collapse of cooperation are possible depending on the magnitude of the mutational variance. Also, making the cost of helping a convex function of the amount of help provided makes it more difficult for cooperative strategies to invade a non-cooperative equilibrium, and for the cooperative equilibrium to resist destabilization by non-cooperative strategies. Finally, we demonstrate that a much greater degree of assortment is required to destabilize a non-cooperative equilibrium in the continuous IPD than in the discrete IPD. The continuous model outlined here suggests that incremental amounts of cooperation lead to rapid decay of cooperation and thus even a large degree of assortment will not be sufficient to allow cooperation to increase when cooperators are rare. The extreme degree of assortment required to destabilize the non-cooperative equilibrium, as well as the instability of the cooperative equilibrium, may help explain why cooperation in Prisoner's Dilemmas is so rare in nature. PMID- 17125799 TI - Cytoprotective and antioxidant role of diallyl tetrasulfide on cadmium induced renal injury: an in vivo and in vitro study. AB - Cadmium (Cd) is an environmental and industrial pollutant that affects various organs in humans and animals. A body of evidence has accumulated implicating the free radical generation with subsequent oxidative stress in the biochemical and molecular mechanisms of Cd toxicity. Since kidney is the critical target of Cd toxicity, we carried out this study to investigate the effects of diallyl tetrasulfide (DTS), an organosulfur compound derived from garlic on Cd induced toxicity in the kidney of rats and also in the kidney cell line (vero cells). In experimental rats, subcutaneous administration of Cd (3 mg/kg bw/day) for 3 weeks induced renal damage, which was evident from significantly increased levels of serum urea and creatinine with significant decrease in creatinine clearance. A markedly increased levels of lipid peroxidation markers (thiobarbituric acid reactive substances and lipid hydroperoxides) and protein carbonyl contents with significant decrease in nonenzymic antioxidants (total sulphydryl groups, reduced glutathione, vitamin C and vitamin E) and enzymic antioxidants (superoxide dismutase, catalase, glutathione peroxidase and glutathione-S-transferase) as well as glutathione metabolizing enzymes (glutathione reductase, and glucose-6 phosphate dehydrogenase) were also observed in Cd intoxicated rats. Coadministration of DTS (40 mg/kg bw/day) and Cd resulted in the reversal of the kidney function accompanied by a significant decrease in lipid peroxidation and increase in the antioxidant defense system. In vitro studies with vero cells showed that incubation of DTS (5-50 microg/ml) with Cd (10 microM) significantly reduced the cell death induced by Cd. DTS at 40 microg/ml effectively blocked the cell death and lipid peroxidation induced by Cd (10 microM) indicating its cytoprotective property. Further, the flow cytometric assessment on the level of intracellular reactive oxygen species using a fluorescent probe 2', 7' dichlorofluorescein diacetate (DCF-DA) confirmed the Cd induced intracellular oxidative stress in vero cells, which was significantly suppressed by DTS (40 microg/ml). The histopathological studies in the kidney of rats also showed that DTS (40 mg/kg bw/day) markedly reduced the toxicity of Cd and preserved the architecture of renal tissue. The present study suggests that the cytoprotective potential of DTS in Cd toxicity might be due to its antioxidant and metal chelating properties, which could be useful for achieving optimum effects in Cd induced renal damage. PMID- 17125800 TI - The source and distribution of polychlorinated dibenzo-p-dioxin and polychlorinated dibenzofurans in sediments of Port Jackson, Australia. AB - Concentrations of polychlorinated dibenzo-p-dioxins (dioxins)(PCDDs) and polychlorinated dibenzofurans (PCDFs)(furans) in sediments from Port Jackson ranged from 32 to 4352 pg WHO-TEQ DF g(-1)dw with a mean of 712 pg g(-1)dw. Maximum total dioxins plus furans concentrations in this sediment was 6,290,000 pg g(-1)dw, which is the highest recorded in Australia and among the highest reported for sediment globally. Permanent bans were placed on fin fishing and prawn trawling in February, 2006 for the whole estuary, based on dioxin tissue burdens. A distinct congener profile corresponded to chemicals known to have been produced by industry on the shores of Homebush Bay in the upper estuary. These chemicals are being dispersed extensively and exceed pre-anthropogenic concentrations in even distant, less developed parts of the harbour. Remediation of sediments is currently being undertaken, with the intention of containing dispersion of dioxins and reducing body burdens in fin fish and prawn populations. PMID- 17125801 TI - Occurrence and fate of polycyclic aromatic hydrocarbons in the coastal surface microlayer. AB - Polycyclic aromatic hydrocarbons (PAHs) were measured in the coastal sea surface microlayer (SML), the sub-surface waters (SSW) and the overlying atmosphere in order to investigate the influence of the SML on contaminant enrichment and air sea exchange. Samples were collected at two contrasting locations of the NW Mediterranean, one urban influenced (off Barcelona, Spain) and another comparatively clean (off Banyuls-sur-Mer, France). Statistical data analysis confirmed the accumulation of PAHs in the SML with respect to the SSW (20.3+/-9.1 vs. 13.1+/-10.0 ng L(-1) in the dissolved phase; 709+/-207 vs. 158+/-111 ng g(-1) in the particulate phase). This accumulation was higher at the contaminated location (Barcelona station) compared with the more pristine one, with PAH enrichments 1.5 and 4.5 times higher for the dissolved and particulate phases, respectively, indicating that the enrichment of PAHs in the SML is dominated by particle transport processes. PMID- 17125802 TI - Geometric criteria for the non-existence of cycles in predator-prey systems with group defense. AB - In this paper, we study the existence of cycles in a predator-prey system in which the prey species is equipped with the group defense capability. Some geometric criteria are developed, relating the location of the two positive equilibria on the prey isocline and the non-existence of cycles. We show that under a general geometric condition, if both positive equilibria lie on a downslope or both lie on an upslope of the prey isocline, cycles do not exist. PMID- 17125803 TI - A model for the interaction between gadoid larvae and their nauplii prey. AB - A continuous model for the growth and death of gadoid larvae, including ecological interactions with their nauplii prey, is examined. The present model has a simpler structure than the model due to Cushing and Horwood [D.H. Cushing, J.W. Horwood, The growth and death of fish larvae. J. Plankt. Res. 16 (3) (1994) 291-300] as it does not explicitly incorporate larval metabolism, although indirectly metabolism is included by means of equations for larval growth. Despite this, the model yields related, although not entirely equivalent, results to those obtained by Cushing and Horwood. In the present model, overcompensation (cf. [W.E. Ricker, Stock and Recruitment, J. Fish. Res. Board. Can. 11 (1954) 559 623]) occurs at limited initial food levels, while at infinite food levels, the recruitment curve becomes monotonically increasing towards an upper limit (cf. [R.J.H. Beverton, S.J. Holt, On the dynamics of exploited fish populations. Fish. Invest. Lond. I 19 (1957)]). Moreover, the present study suggests that the duration of the larval stage, the metamorphosis time tau is highly important to the recruitment process, in accordance with Cushing and Horwood. When food is limited the metamorphosis is delayed, causing the larval population to experience (density dependent) mortality for a sufficient long time to make the recruitment curve overcompensatory. It is not necessarily the desire to derive a particular formula for the recruitment curve, as this is probably impossible anyway, except for particular examples. However, reduced versions of the model that in some sense are close to the original model, are examined, and it is argued that many general features of the general model are retained in such examples. PMID- 17125804 TI - Effects of tick population dynamics and host densities on the persistence of tick borne infections. AB - The transmission and the persistence of tick-borne infections are strongly influenced by the densities and the structure of host populations. By extending previous models and analysis, in this paper we analyse how the persistence of ticks and pathogens, is affected by the dynamics of tick populations, and by their host densities. The effect of host densities on infection persistence is explored through the analysis and simulation of a series of models that include different assumptions on tick-host dynamics and consider different routes of infection transmission. Ticks are assumed to feed on two types of host species which vary in their reservoir competence. Too low densities of competent hosts (i.e., hosts where transmission can occur) do not sustain the infection cycle, while too high densities of incompetent hosts may dilute the competent hosts so much to make infection persistence impossible. A dilution effect may occur also for competent hosts as a consequence of reduced tick to host ratio; this is possible only if the regulation of tick populations is such that tick density does not increase linearly with host densities. PMID- 17125805 TI - Roscovitine differentially affects CaV2 and Kv channels by binding to the open state. AB - Roscovitine potently inhibits cyclin-dependent kinases (CDK) and can independently slow the closing of neuronal (CaV2.2) calcium channels. We were interested if this drug could affect other ion channels similarly. Using whole cell recordings, we found that roscovitine specifically slows deactivation of all CaV2 channels (N, P/Q and R) by binding to the open state. This effect had a rapid onset and EC(50)=54, 120 and 54microM for N-, P/Q-, and R-type channels, respectively. Deactivation of other channel types was not slowed, including L type calcium channels (CaV1.2, CaV1.3), potassium channels (native, Kv4.2, Kv2.1 and Kv1.3), and native sodium channels. However, most of the channels tested were inhibited by roscovitine. The inhibition was characterized by slow development and a lower affinity (EC(50)=100-300microM). Surprisingly, potassium channels were rapidly inhibited with an EC(50)=23microM, which is similar to the EC(50) for roscovitine block of cell division [Meijer, L., Borgne, A., Mulner, O., Chong, J., Blow, J., Inagaki, N., Inagaki, M., Delcros, J., Moulinoux, J., 1997. Biochemical and cellular effects of roscovitine, a potent and selective inhibitor of the cyclin-dependent kinases cdc2, cdk2 and cdk5. Eur. J. Biochem. 243, 527 536]. Potassium current inhibition seemed to result from open channel block. The high potency of these two rapid onset effects makes them complicating factors for ongoing clinical trials and research using roscovitine. Thus, the physiology and pharmacology of slow CaV2 deactivation and potassium channel block must be explored. PMID- 17125806 TI - Differences in kinetics of structurally related competitive GABA(A) receptor antagonists. AB - Previously, 4-alkyl and 4-aryl substituted analogues of the low-efficacy partial GABA(A) receptor agonist 5-(4-piperidyl)-3-isothiazole (4-PIOL) have been identified as competitive GABA(A) receptor antagonists. These structurally related competitive antagonists show marked differences in their kinetic properties. The kinetics of 20 4-alkyl and 4-aryl substituted analogues of 4 PIOL, two 4-arylalkyl substituted 3-isothiazolol analogues and the classical GABA(A) receptor antagonist SR95531 was studied in cultured cerebral cortical neurons using whole-cell patch-clamp techniques. The kinetics of the antagonists was studied indirectly by measuring the changes in the response of the full GABA(A) receptor agonist isoguvacine (IGU) induced by concurrent application of an antagonist. When added, the majority of the antagonists did not affect the rate of deactivation of the IGU-induced responses. When removed, however, the majority of the antagonists slowed the reactivation phase of IGU implying that the dissociation of the antagonist from the GABA(A) receptor is the rate-limiting step. Surprisingly, the functional off-rates of the antagonists seemed to correlate better with the lipophilicity of the compounds than with the affinity and potency. This suggests that the dissociation of the tested antagonists from the GABA(A) receptor is restricted by lipophilic interactions, perhaps with the aromatic amino acids surrounding the GABA binding site. PMID- 17125807 TI - Bilingualism as a protection against the onset of symptoms of dementia. AB - This study examined the effect of lifelong bilingualism on maintaining cognitive functioning and delaying the onset of symptoms of dementia in old age. The sample was selected from the records of 228 patients referred to a Memory Clinic with cognitive complaints. The final sample consisted of 184 patients diagnosed with dementia, 51% of whom were bilingual. The bilinguals showed symptoms of dementia 4 years later than monolinguals, all other measures being equivalent. Additionally, the rate of decline in Mini-Mental State Examination (MMSE) scores over the 4 years subsequent to the diagnosis was the same for a subset of patients in the two groups, suggesting a shift in onset age with no change in rate of progression. PMID- 17125808 TI - Unrevealed structural requirements for auxin-like molecules by theoretical and experimental evidences. AB - An computational-biostatistical approach, supported by ab initio optimizations of auxin-like molecules, was used to find biologically meaningful relationships between quantum chemical variables and fresh bioassay's data. It is proven that the auxin-like recognition requires different molecular assembling states. We suggest that the carboxyl group is not the determining factor in explaining the biological auxin-like conduct. The biological effects depends essentially on the chemical condition of the ring system. The aim to find active molecules (quantum objects) via statistical grouping-analysis of molecular quantum similarity measures was verified by bioactivity assays. Next, this approach led to the discovery of a non-carboxylated active auxin-like molecule (2,6-dibromo-phenol). This is the first publication on structure activity relationship of auxin-like molecules, which relies on highly standardized bioassays of different auxins screened in parallel as well as analysed by multi-dimensional scaling. PMID- 17125809 TI - Factors influencing drug compliance in the mass drug administration programme against filariasis in the Western province of Sri Lanka. AB - This descriptive, comparative, cross-sectional, randomized community-based study was done to determine factors influencing drug compliance in the mass drug administration (MDA) programme against filariasis in selected urban and rural populations within the Western province of Sri Lanka. The study population was selected using the cluster sampling method. Factors influencing drug compliance were determined by administration of an interviewer-based pre-tested questionnaire eliciting information regarding drug compliance, socio-economic status, educational background, knowledge, attitudes and practices with regard to the 2004 MDA. A total of 2319 people aged between 10 and 90 years (median 40) responded to the questionnaire. The belief that the MDA programme was beneficial was the most important factor affecting drug compliance, as revealed by multivariate analysis of the combined populations (P<0.001). This was so even in the urban population (P<0.001), while the belief regarding the severity (danger) of filariasis was important in the rural population (P=0.013), when the areas were considered individually. Therefore, it is essential for awareness programmes to highlight the dangers (complications) of the disease and to influence the community to perceive the benefits of a filariasis-free community as well as 'beyond filariasis' benefits of having albendazole given in combination in the MDA programme. PMID- 17125810 TI - Trichostatin A, a critical factor in maintaining the functional differentiation of primary cultured rat hepatocytes. AB - Histone deacetylase inhibitors (HDI) have been shown to increase differentiation related gene expression in several tumor-derived cell lines by hyperacetylating core histones. Effects of HDI on primary cultured cells, however, have hardly been investigated. In the present study, the ability of trichostatin A (TSA), a prototype hydroxamate HDI, to counteract the loss of liver-specific functions in primary rat hepatocyte cultures has been investigated. Upon exposure to TSA, it was found that the cell viability of the cultured hepatocytes and their albumin secretion as a function of culture time were increased. TSA-treated hepatocytes also better maintained cytochrome P450 (CYP)-mediated phase I biotransformation capacity, whereas the activity of phase II glutathione S-transferases (GST) was not affected. Western blot and qRT-PCR analysis of CYP1A1, CYP2B1 and CYP3A11 protein and mRNA levels, respectively, further revealed that TSA acts at the transcriptional level. In addition, protein expression levels of the liver enriched transcription factors (LETFs) hepatic nuclear factor 4 alpha (HNF4alpha) and CCAAT/enhancer binding protein alpha (C/EBPalpha) were accordingly increased by TSA throughout culture time. In conclusion, these findings indicate that TSA plays a major role in the preservation of the differentiated hepatic phenotype in culture. It is suggested that the effects of TSA on CYP gene expression are mediated via controlling the expression of LETFs. PMID- 17125811 TI - Human papillomavirus (HPV) types 101 and 103 isolated from cervicovaginal cells lack an E6 open reading frame (ORF) and are related to gamma-papillomaviruses. AB - Complete genomes of HPV101 and HPV103 were PCR amplified and cloned from cervicovaginal cells of a 34-year-old female with cervical intraepithelial neoplasia grade 3 (CIN 3) and a 30-year-old female with a normal Pap test, respectively. HPV101 and HPV103 contain 4 early genes (E7, E1, E2, and E4) and 2 late genes (L2 and L1), but both lack the canonical E6 ORF. Pairwise alignment similarity of the L1 ORF nucleotide sequences of HPV101 and HPV103 indicated that they are at least 30% dissimilar to each other and all known PVs. However, similarities of the other ORFs (E7, E1, E2, and L2) indicated that HPV101 and HPV103 are most related to each other. Phylogenetic analyses revealed that these two types form a monophyletic clade, clustering together with the gamma- and pi PV groups. These data demonstrated that HPV genomes closely related to papillomaviruses identified from cutaneous epithelia can be isolated from the genital mucosal region. Moreover, this is the first report of HPVs lacking an E6 ORF and phylogenetic evidence suggests this occurred subsequent to their emergence from the gamma-/pi-PVs. PMID- 17125812 TI - Confronting the realities of wastewater aquaculture in peri-urban Kolkata with bioeconomic modelling. AB - Wastewater reuse for aquaculture is a reality in several Asian countries, however, traditional practices face constraints including inadequate or contaminated wastewater inputs and growing concern over health risks. Based on Kolkata and a wastewater flow of 550,000m(3)d(-1), rational and conventional designs for lagoon-based wastewater treatment and reuse through aquaculture were compared using bioeconomic modelling. Outcomes showed the rational design required a larger area than the conventional or traditional systems, but that financial returns, nutrient retention and fish production were higher; gross fish production employing rational and conventional designs was 45,500 and 11,560t, respectively. However, limited land availability and constraints to reconfiguring the existing system make implementation of the rational design unlikely. Findings suggest traditional practices could be enhanced by adopting wastewater treatment prior to reuse, modifying fish production strategies, and monitoring to safeguard health. Bioeconomic modelling constitutes a useful tool in comparing treatment and reuse options, permitting the sensitivity of financial returns to changing costs and recent revisions to WHO guidelines for safe wastewater reuse to be assessed, however, social and environmental consequences demand consideration. PMID- 17125813 TI - Bioavailability and degradation of phenanthrene in compost amended soils. AB - Bioavailability in soil of organic xenobiotics such as phenanthrene is limited by mechanisms of diffusion of the xenobiotics within soil micropores and organic matter. The agricultural utilization of compost may reduce the risk connected to organic xenobiotic contamination by means of: (i) a reduction of the bioavailable fraction through an increased adsorption and (ii) an enhanced degradation of the remaining bioavailable fraction through an inoculum of degrading microorganisms. Aim of this work is to test this hypothesis by assessing the effects of compost amendment on the bioavailability and degradation of phenanthrene in soil. Experiments were carried out in both sterilized and non-sterilized conditions, and chemical and microbiological analyses were carried out in order to determine the extent of degradation and bioavailability and to monitor the evolution of the soil micro flora in time. Bioavailability was assessed in sterilized microcosms, in order to assess the physical effects of compost on aging processes without the influence of microbial degradation. Results showed that bioavailability is significantly reduced in soils amended with compost, although no differences were found at the 2 doses of compost studied. In non-sterilized soils the amount of phenanthrene degraded was always higher in the amended soils than in the non amended one. Microbiological analyses confirmed the presence of a higher number of phenanthrene degraders in the amended soils and in samples of compost alone. These results suggest that compost induces the degradation in soils of easily degradable compounds such as phenanthrene, when the proper bacteria are in the compost; more resistant xenobiotics may instead be trapped by the compost organic matter, thus becoming less available. PMID- 17125814 TI - Changes in the chemical composition of water-extractable organic matter during composting: distribution between stable and labile organic matter pools. AB - Aerobic decomposition and stabilization of organic matter during the composting of waste materials is primarily due to the biochemical transformation of water soluble compounds in the liquid phase by the microbial biomass. For this reason water-soluble organic matter represents the most active fraction of compost, both biologically and chemically, and thus should directly reflect the biochemical alteration of organic matter. This work aims to elucidate the microbial-mediated processes responsible for the distribution of soluble organic matter between stable and labile pools with composting time. Accordingly, chemical analysis as well as UV absorption, and 1H and 13C-NMR spectroscopy of samples collected during the industrial composting of urban waste revealed microbial induced transformation of water-extractable organic matter over time. The chemical composition changed from labile, hydrophilic, plant-derived organic compounds in the beginning to predominately stable, hydrophobic moieties comprising lignin derived phenols and microbially-derived carbohydrates at later stages of composting. PMID- 17125815 TI - Transplantation and regeneration in the heart of the Mediterranean. AB - The second half of the 20th century witnessed the birth of organ transplantation, and failing organs can now be replaced with healthy ones procured from living or cadaveric donors, allowing their recipient to start, or return to, an active life. Major milestones in the field were set in the eighties and nineties at the University of Pittsburgh Medical Center (UPMC), an institution that made it a mission to spread its expertise internationally. A successful partnership between UPMC and the Region of Sicily gave rise to the Mediterranean Institute for Transplantation and Highly Specialized Therapies (ISMETT), the only Italian facility entirely dedicated to transplantation of all solid organs and therapies for the treatment of end-stage organ failure. In its first seven years of activity, ISMETT has become a major referral center for patients from the entire Mediterranean Basin and the Middle East. Despite the fact that organ transplantation is the current gold standard for end-stage organ failure, the field is facing a worldwide emergency represented by the chronic shortage of organ donors. Research aimed at understanding the molecular networks involved in organ-specific ageing and their relationship with maintenance networks and organ failure should be actively encouraged and supported as it could ultimately allow to control organ performance and lifespan, increasing the number of organs available for transplant. PMID- 17125816 TI - Age-related cardiac disease model of Drosophila. AB - We have begun to study the genetic basis of deterioration of cardiac function in the fruit fly Drosophila melanogaster as an age-related cardiac disease model. For this purpose we have developed heart function assays in Drosophila and found that the fly's cardiac performance, as that of the human heart, deteriorates with age: aging fruit flies exhibit a progressive increase in electrical pacing induced heart failure as well as in arrhythmias. The insulin receptor and associated pathways have a dramatic and heart-autonomous influence on age-related cardiac performance in flies, suggestive of potentially similar mechanisms in regulating cardiac aging in vertebrates. Compromised KCNQ and K(ATP) ion channel functions also seem to contribute to the decline in heart performance in aging flies, suggesting that the corresponding vertebrate gene functions may similarly decline with age, in addition to their conserved role in protecting against arrhythmias and hypoxia/ischemia, respectively. The fly heart is thus emerging as a promising genetic model for studying the age-dependent decline in organ function. PMID- 17125817 TI - Sampling challenges posed by continental-scale soil landscape modeling. AB - Limited research exists to guide the development of continental-scale field sampling approaches. We developed a method to compare different sampling schemes designed for continental-scale environmental analysis. Using simulated annealing, we produced hypothetical soil property surfaces for the conterminous United States. The surfaces exhibited varying degrees of polygonality and conditioning by existing soil datasets. Differences in mean between a gridded sample and a polygon-centered sample were evaluated using randomization and other statistical tests. We saw a weak evidence in support of the hypothesis that if a polygonal structure exists in a surface being sampled, then it should be utilized during the actual sampling routine. We saw a somewhat stronger evidence for the hypothesis that differences in local sample density across a continental trend surface can lead to significant sampling bias. Ultimately, the success of different sampling strategies designed for use in continental-scale environmental analysis will largely depend on the ability to anticipate the spatial variability of the variable being measured. PMID- 17125818 TI - Serotonin (5-hydroxytryptamine, 5-HT) immunoreactive endocrine and neural elements in the chromaffin enteropancreatic system of amphibians and reptiles. AB - The diffuse chromaffin enteropancreatic system of nine species of amphibians (newts, frogs) and reptiles (turtles, lizards, snakes) was investigated immunohistochemically for the presence and topographic distribution of serotonin (5-hydroxytryptamine, 5-HT). The study revealed various numbers of serotonin producing cells in the pancreas and intestinal epithelium and also immunolabelled nerve profiles in the villi of all species studied. In addition, two different morphological populations of serotonin cells ("open" and "closed") were localized in the functional segments of the intestines in the representative species of all the taxa investigated. Semi-quantitative evaluation of the immunolabelled pancreatic and enteric cells revealed significantly different mean numbers of labelled cells in different amphibian and reptilian taxa, and also between the various successive gut segments of each taxon. The ratio between "open" and "closed" varieties of serotonin cells recorded along the intestines followed a decreasing trend, progressive in lizards and snakes and more abrupt in newts, frogs and turtles. The above findings may help resolve several key stages of the phylogenetic evolution of poikilothermic vertebrates. PMID- 17125820 TI - Basophil FcepsilonRI histamine release parallels expression of Src-homology 2 containing inositol phosphatases in chronic idiopathic urticaria. AB - BACKGROUND: Basophils are implicated in the pathogenesis of chronic idiopathic urticaria (CIU). Autoantibodies to the IgE receptor (FcepsilonRI) and serum histamine releasing activity have been detected in some subjects with CIU, although their role in vivo is unclear. Basophils of patients with CIU have altered FcepsilonRI-mediated histamine release (HR); however, the mechanism is unknown. In the basophil FcepsilonRI signaling pathway, protein levels of Src homology 2-containing-5'-inositol phosphatase (SHIP)-1 are inversely correlated with the release of mediators or releasability. A related phosphatase, SHIP-2, is a negative regulator of monocyte IgG receptor (FcgammaR) signaling . We hypothesized that SHIP levels are altered in CIU basophils. METHODS: Blood basophils were isolated from cold urticaria, CIU, or normal donors, and FcepsilonRI-dependent and independent HR were quantified. Protein levels of SHIP 1, SHIP-2, spleen tyrosine kinase, and phosphorylated Akt were determined by Western blotting. Subjects' serum was tested for serum histamine releasing activity and anti-FcepsilonRIalpha antibodies. RESULTS: CIU basophils displayed a bimodal response to anti-IgE activation. One half of CIU subjects' basophils had reductions in anti-IgE-induced HR and were designated nonresponders (CIU NR). CIU NR basophil HR remained diminished at 10-fold to 30-fold higher doses of anti IgE. CIU anti-IgE responder basophils had HR similar to normal subjects. SHIP-1 and SHIP-2 proteins were increased in CIU NR basophils and were linked to reduced phosphoAkt after anti-IgE stimulation. CIU basophil anti-IgE response was not related to the presence of serologic factors. CONCLUSION: In CIU basophils, the observed changes in FcepsilonRI signaling pathway molecule expression may underlie changes in releasability. CLINICAL IMPLICATIONS: Patients with CIU can be segregated on the basis of basophil functional phenotype. PMID- 17125821 TI - Life-threatening anaphylaxis to kiwi fruit: protective sublingual allergen immunotherapy effect persists even after discontinuation. PMID- 17125822 TI - Childhood Asthma Management Program: lessons learned. AB - The National Heart, Lung, and Blood Institute Childhood Asthma Management Program was initiated in 1991 and is now the largest and most comprehensive study of long term intervention with anti-inflammatory therapy in children with mild to moderate asthma. The purpose of this perspective is to review key findings of the study and lessons learned in conducting research in more than 1000 children with persistent asthma for more than 10 years. A key lesson was absence of a continued effect of inhaled corticosteroid on lung growth during long-term follow-up even as symptoms and airway responsiveness remained improved. PMID- 17125823 TI - Mannan-binding lectin in young children with asthma differs by level of severity. PMID- 17125824 TI - The neural underpinnings of asthma. PMID- 17125825 TI - Asthma treatment and asthma prevention: a tale of 2 parallel pathways. AB - Three recent clinical trials used different study designs to test the hypothesis that early introduction of inhaled corticosteroids in infants and young children at high risk for the development of asthma could change the natural course of the disease. All 3 trials reached the same conclusion: treatment requirement, symptom frequency while off treatment, and lung function did not differ between children receiving active drug or placebo, with outcomes measured 2 to 4 years after randomization. These findings challenge the concept that the inflammatory processes that cause asthma symptoms and are responsive to inhaled corticosteroids are also responsible for the chronic changes in airway structure and function that are believed to predispose to the development of persistent asthma. This conclusion is supported by studies showing that bronchial hyperresponsiveness, independent of current asthma symptoms, is associated with subsequent deficits in airway function growth during childhood. Successful strategies for the prevention of asthma will require a better understanding of the genetic, environmental, and developmental factors that predispose toward inappropriate responses to airway injury. Abnormal airway remodeling and persistent dysregulation of airway tone might be the final common pathway for different disease mechanisms, and this might explain the heterogeneity of clinical phenotypic syndromes that go under the common label of "asthma." PMID- 17125826 TI - Significant association of FcepsilonRIalpha promoter polymorphisms with aspirin intolerant chronic urticaria. AB - BACKGROUND: Although the mechanism that underlies aspirin hypersensitivity is not completely understood, an IgE-mediated response was reported for a patient with aspirin-intolerant chronic urticaria (AICU). OBJECTIVE: We investigated whether genetic polymorphisms on the alpha-chain of the high-affinity IgE receptor (FcepsilonRIalpha) gene were associated with the AICU phenotype. METHODS: We genotyped 2 promoter polymorphisms (-344C>T and -95T>C) of FcepsilonRIalpha gene in the Korean population, and the functional effect of the -344C>T polymorphism was analyzed by using a luciferase reporter assay and an electrophoretic mobility shift assay. RESULTS: The rare allele frequency of the -344C>T polymorphism was significantly higher in the patients with AICU compared with the other subjects (P= .008 for AICU vs aspirin-tolerant chronic urticaria; P= .03 for AICU vs controls). This polymorphism was also significantly associated with total serum IgE concentrations and a higher rate of atopy in the patients with AICU (P= .01 and .05, respectively). The reporter plasmid that carried the -344T allele exhibited significantly higher promoter activity in a rat mast cell line (RBL 2H3) compared with the promoter activity of the -344C allele (P< .001). We found that transcription factor Myc-associated zinc finger protein preferentially bound the -344C promoter. Moreover, patients with AICU with the heterozygous CT genotype of the -344C>T polymorphism exhibited greater anti-IgE-mediated histamine release compared with those with the homozygous CC genotype. CONCLUSION: These results suggest that the -344C>T polymorphism of the FcepsilonRIalpha promoter may be associated with increased expression of FcepsilonRIalpha on mast cells and enhanced release of histamine. CLINICAL IMPLICATIONS: The FcepsilonRIalpha -344C>T polymorphism may contribute to the development of AICU. PMID- 17125827 TI - Differentiation of monocytes into macrophages induces the upregulation of histamine H1 receptor. AB - BACKGROUND: Histamine modulates several functions in human monocytes, macrophages, and dendritic cells. However, responses elicited by histamine differ depending on cell type, suggesting variable expression of histamine receptors in the monocyte/macrophage lineage. OBJECTIVE: We sought to examine whether the expression of H(1) receptors was regulated by cell differentiation of human monocytes into macrophages or dendritic cells. METHODS: Expression of H(1) receptor was evaluated by RT-PCR and western blot in monocytes, monocyte-derived macrophages (MDMs), monocyte-derived dendritic cells (DCs) and human lung macrophages (HLMs). RESULTS: Expression of H(1) receptor mRNA and protein was higher in HLMs and DCs than in monocytes. H(1) expression was approximately 15 fold and 4-fold higher in MDMs and HLMs, respectively, as compared with that seen in monocytes. H(1) receptor protein was undetectable in monocytes, whereas it was conspicuous in MDMs. Simultaneous analysis of H(2) and H(1) mRNA expression indicated that the H(2)/H(1) ratio decreased from 202.7 +/- 14.8 in monocytes to 2.2 +/- 0.4 in MDM and 39.5 +/- 5.0 in DCs. Incubation of monocytes with histamine neither affected intracellular Ca(2+) concentrations nor influenced IL 8 production. In contrast, histamine rapidly induced a Ca(2+) signal and stimulated IL-8 production in MDMs. Both effects were inhibited by H(1) blockade with levocetirizine, but not by H(2) blockade with ranitidine. CONCLUSIONS: Differentiation of monocytes into macrophages or dendritic cells is associated with profound changes of histamine receptor expression. Upregulation of H(1) receptors confers on macrophages the capacity of being activated by histamine. CLINICAL IMPLICATIONS: Regulation of H(1) and H(2) receptor expression in the monocyte/macrophage lineage can be relevant to the pathogenesis of allergic inflammation. PMID- 17125828 TI - The parthenogenetic activation of canine oocytes with Ca-EDTA by various culture periods and concentrations. AB - In the present study, canine oocytes were exposed to various concentrations of and durations of exposure to EDTA saturated with Ca(2+) (Ca-EDTA), a cell membrane-impermeable metal ion chelator, to determine if parthenogenetic activation could be induced. When oocytes were cultured for 48 or 72 h in parthenogenetic activation medium (PAM) without Ca-EDTA (control) or PAM supplemented with 1 or 5mM Ca-EDTA, the highest rate of pronuclear formation (PN) was obtained in oocytes cultured in 1mM Ca-EDTA for 48 h (8.0%; P<0.05). There was no pronuclear formation in the control group (PAM without Ca-EDTA). Oocytes treated with 5mM Ca-EDTA for 48 h or 1mM Ca-EDTA for 72 h formed a parthenogenetic pronucleus (3.1 and 4.5, respectively). However, there was no pronuclear formation in oocytes treated with 5mM Ca-EDTA for 72 h. In summary, exposure to Ca-EDTA can induce pronuclear formation in canine oocytes. PMID- 17125829 TI - Long-term TEM analysis of the nanoleakage patterns in resin-dentin interfaces produced by different bonding strategies. AB - OBJECTIVES: The aim of this study was to evaluate the ability of etch-and-rinse and self-etching adhesive systems to prevent time- and water-induced nanoleakage in resin-dentin interfaces over a 6-month storage period. METHODS: Five commercial adhesives were tested, which comprise three different strategies of bonding resins to tooth hard tissues: one single-step self-etching adhesive (One up Bond F (OB), Tokuyama); two two-step self-etching primers (Clearfil SE Bond (SE) and an antibacterial fluoride-containing system, Clearfil Protect Bond (CP), Kuraray Inc.); two two-step etch-and-rinse adhesives (Single Bond (SB), 3M ESPE and Prime&Bond NT (PB), Dentsply). Restored teeth were sectioned into 0.9 mm thick slabs and stored in water or mineral oil for 24 h, 3 or 6 months. A silver tracer solution was used to reveal nanometer-sized water-filled spaces and changes that occurred over time within resin-dentin interfaces. Characterization of interfaces was performed with the TEM. RESULTS: The two two-step self-etching primers showed little silver uptake during the 6-month experiment. Etch-and-rinse adhesives exhibited silver deposits predominantly within the hybrid layer (HL), which significantly increased for SB after water-storage. The one-step self etching adhesive OB presented massive silver accumulation within the HL and water trees protruding into the adhesive layer, which increased in size and quantity after water-storage. After storage in oil, reduced silver deposition was observed at the interfaces for all groups. SIGNIFICANCE: Different levels of water-induced nanoleakage were observed for the different bonding strategies. The two-step self etching primers, especially the antibacterial fluoride-containing system CP, showed the least nanoleakage after 6 months of storage in water. PMID- 17125830 TI - Novel light-cured resins and composites with improved physicochemical properties. AB - OBJECTIVES: The aim of this study was to determine the effect of two new diluent agents (Bis-GMA analogues), at different dilution levels and filler contents on relevant physicochemical properties of several novel resins and composites containing Bis-GMA as matrix. Composites using TEGDMA as diluent were used as control. METHODS: Twenty formulations were prepared combining three monomer mixtures (Bis-GMA/TEGDMA, Bis-GMA/CH(3) Bis-GMA and Bis-GMA/CF(3) Bis-GMA), at three dilution levels (85/15, 10/90, 0/100) and two percentages of filler loading (silanated barium aluminosilicate glass): 0%, 10%, 35%. Preliminary rheological testing was performed in order to obtain the viscosity of the resin samples. Resins and composites were then inserted into molds and light-cured (500mW/cm(2)). The properties evaluated were: (1) homogeneity of curing (HC), using FTIR or Vickers microindentor, (2) microhardness, by a Vickers microindentor, (3) depths of cure and oxygen inhibitor effect (OIE), quantified by scraping, (4) water contact angle on the materials surface, (5) water sorption and solubility, performed by the Oysaed-Ruyter method and (6) scanning electron microscopy analysis of the specimens surfaces. Data were analyzed by ANOVA and Student-Newman-Keuls tests (p<0.05). RESULTS: Materials with CH(3) Bis-GMA and CF(3) Bis-GMA exhibited less hydrophilicity, water sorption and solubility. Bis GMA dilution induced an increase in depth of cure and promoted a higher OIE, particularly when the diluent was TEGDMA. Filler loading reduced the OIE and increased hydrophobicity of the resins. SIGNIFICANCE: CH(3) Bis-GMA may be considered as good candidate to be used as diluent because when replacing TEGDMA induced lower hydrolytic degradation and increase in HC. PMID- 17125831 TI - Effects of sterilisation method on surface topography and in-vitro cell behaviour of electrostatically spun scaffolds. AB - Electrostatic spinning is a potentially significant technique for scaffold production within the field of tissue engineering; however, the effect of sterilisation upon these structures is not known. This research investigated the extent of any topographical alteration to electrostatically spun scaffolds post production through sterilisation, and examined any subsequent effect on contacting cells. Scaffolds made from Tecoflex SG-80A polyurethane were sterilised using ethylene oxide and UV-ozone. Scaffold topography was characterized in terms of inter-fibre separation (ifs), fibre diameter (f.dia) and surface roughness. Cell culture was performed over 7 days with both mouse L929 and human embryonic lung fibroblasts, the results of which were assessed using SEM, image analysis and confocal microscopy. Sterilisation by UV-ozone and ethylene oxide decreased ifs and increased f.dia; surface roughness was decreased by UV-ozone but increased by ethylene oxide. Possible mechanisms to explain these observations are discussed, namely photo-oxidative degradation in the case of UV ozone and process-induced changes in surface roughness. UV-ozone sterilised scaffolds showed greater cell coverage than those treated with ethylene oxide, but lower coverage than all the controls. Changes in cell attachment and morphology were thought to be due to the changes in topography brought about by the sterilisation process. We conclude that surface modification by sterilisation could prove to be a useful tool at the final stage of scaffold production to enhance cell contact, phenotype or function. PMID- 17125832 TI - Long circulating lipid nanocapsules for drug detoxification. AB - Uncoated and poly(ethylene glycol) (PEG)-decorated lipid nanocapsules (NC) prepared from medium chain triglycerides were investigated both in vitro and in vivo as parenteral detoxifying colloids for their ability to sequester haloperidol, docetaxel and paclitaxel. In vitro studies showed that the uptake depended on the nature of the drug and the composition of NC core and shell. In the case of haloperidol, maximal affinity was achieved upon incorporation of a complexing fatty acid. In plasma lipoprotein distribution studies, the association of both haloperidol and docetaxel into triglyceride-rich lipoprotein fraction was significantly increased in the presence of NC. The ability of the NC to lower the free drug concentrations in incubation medium was confirmed by cytotoxicity studies, where the antiproliferative activity of docetaxel was significantly decreased in the presence of NC. Using docetaxel as drug model, the NC were finally evaluated for their uptake potential in mice by one of the following administration sequences between the drug solution (Taxotere, DTX) and NC: NC-DTX, PEG(NC)-DTX and DTX-PEG(NC). Irrespective of the administration sequence, the NC increased the blood levels of docetaxel due to the in situ sequestration of drug by the circulating carrier. These findings suggest that lipid NC could be used as a non-specific mode to deal with the sequestration of molecules with high affinity for oils. PMID- 17125833 TI - Labour induces increased concentrations of biglycan and hyaluronan in human fetal membranes. AB - OBJECTIVE: The proteoglycan decorin stabilizes collagen whereas biglycan and hyaluronan disrupt well-organized collagen. The aim was to compare hyaluronan and proteoglycans in human fetal membranes obtained before and after spontaneous labour at term. STUDY DESIGN: Prelabour samples of fetal membranes (N=9) were obtained from elective caesarean sections and regionally sampled from over the cervix (cervical membranes) and mid-zone samples between this area and the placental edge. Postlabour samples (N=11) were obtained from spontaneous vaginal delivery and also regionally sampled. Amnion and chorio-decidua were analysed separately. The proteoglycans decorin and biglycan were analysed using alcian blue precipitation, SDS polyacrylamide gel electrophoresis and immunostaining. Hyaluronan was analysed using a radioimmunoassay and by histochemistry. Collagen was measured by estimating hydroxyproline content. RESULTS: In prelabour membranes the biglycan concentration (microg/mg wtw) in the cervical amnion was 40% lower than in the mid-zone amnion (P<0.05). After delivery the cervical amnion showed a twofold increase in biglycan (P<0.05), a 30% decrease in collagen (P<0.05), and a 50% decrease in decorin concentration (P<0.05). In mid-zone samples after delivery the concentrations of hyaluronan showed an increase form 1.0 to 4.9 microg/mg wtw (P<0.05). Histology demonstrated a gelatinous substance, which separated amnion and chorio-decidua, in particular at the cervical site. This gelatinous substance contained hyaluronan at a concentration of 3.0 microg/mg wtw. CONCLUSION: It is well established that prelabour fetal membranes are considerably stronger than postlabour fetal membranes. Two features may explain this; a weakening of the amnion combined with a separation of amnion and chorio-decidua. The biomechanical changes are consistent with the decrease in collagen and decorin, and the increase in hyaluronan and biglycan demonstrated in this study. The separation of the membranes is caused by the formation of a gelatinous substance, rich in hyaluronan. The results indicate that the biomechanical changes are not merely secondary to the stress of labour but that an active maturation process is involved. PMID- 17125835 TI - Efficacy of conventional cytogenetics and FISH for EGR1 to detect deletion 5q in hematological disorders and to assess response to treatment with Lenalidomide. AB - In clinical practice, whether FISH for EGR1 in interphase nuclei has similar efficacy to detect deletion 5q anomalies as conventional cytogenetic studies is unknown. We compared conventional cytogenetics and FISH for 145 patients with deletion 5q and detected this anomaly by both methods in 144. Nine patients with myelodysplasia were studied before and after treatment with Lenalidomide and results were concordant for 28 of 29 specimens. FISH did not detect anomalies other than deletion 5q in 31 patients. This study suggests FISH is useful to detect deletion 5q, but is not a substitute for conventional cytogenetics. PMID- 17125834 TI - Ca2+ signal summation and NFATc1 nuclear translocation in sympathetic ganglion neurons during repetitive action potentials. AB - NFATc-mediated gene expression constitutes a critical step during neuronal development and synaptic plasticity. Although considerable information is available regarding the activation and functionality of specific NFATc isoforms, in neurons little is known about how sensitive NFAT nuclear translocation is to specific patterns of electrical activity. Here we used high-speed fluo-4 confocal imaging to monitor action potential (AP)-induced cytosolic Ca2+ transients in rat sympathetic neurons. We have recorded phasic and repetitive AP patterns, and corresponding Ca2+ transients initiated by either long (100-800 ms) current-clamp pulses, or single brief (2 ms) electrical field stimulation. We address the functional consequences of these AP and Ca2+ transient patterns, by using an adenoviral construct to express NFATc1-CFP and evaluate NFATc1-CFP nuclear translocation in response to specific patterns of electrical activity. Ten Hertz trains stimulation induced nuclear translocation of NFATc1, whereas 1 Hz trains did not. However, 1 Hz train stimulation did result in NFATc1 translocation in the presence of 2 mM Ba2+, which inhibits M-currents and promotes repetitive firing and the accompanying small (approximately 0.6 DeltaF/F0) repetitive and summating Ca2+ transients. Our results demonstrate that M-current inhibition mediated spike frequency facilitation enhances cytosolic Ca2+ signals and NFATc1 nuclear translocation during trains of low frequency electrical stimulation. PMID- 17125836 TI - Experimental evaluation of the usefulness of feathers as a non-destructive biomonitor for polychlorinated biphenyls (PCBs) using silastic implants as a novel method of exposure. AB - Risk assessment of pollutants requires both monitoring studies in the field and experimental exposure studies. In this study, we evaluated silastic implants as an alternative method of exposure for use in toxicological studies and at the same time evaluated the usefulness of feathers as a non-destructive biomonitor for PCBs. European starlings (Sturnus vulgaris) were exposed to different doses (including a control group) of environmentally relevant concentrations of PCB 153 during a 15-week period using silastic implants with both ends/only one end sealed. After implantation, there was a rapid and significant increase in PCB 153 blood concentration in the exposed groups. The significant differences in blood concentrations among the treatment groups show that silastic implants are useful as a method of exposure. Moreover, the ratio between the tissue concentrations of two treatment groups reflected the difference in implantation doses between these groups. There was also a clear difference in tissue concentrations among the treatment groups, although we could not test this statistically due to the small sample sizes. The slow release kinetics for a prolonged period and the relatively stable blood concentrations during the 15-week period render silastic tubes very interesting to study the effects of chronic exposure to pollutants. Our results also revealed that sealing both ends of the implant instead of only one did not significantly affect the exposure. There were strong, significant positive correlations between the blood and the tissues, which confirm the use of blood to monitor PCBs. To evaluate the usefulness of feathers as a non-destructive biomonitor for PCBs, we plucked the original and newly grown wing and tail feathers. We observed strong, significant positive correlations between the concentrations in the newly grown feathers and concentrations in the muscle, liver, brain and blood. PCB 153 concentrations in the newly grown feathers differed among the treatment groups. To our knowledge, our results provide the first experimental evidence that feathers are useful as a non-destructive biomonitoring tool for PCBs. PMID- 17125837 TI - A novel domain antibody rationally designed against TNF-alpha using variable region of human heavy chain antibody as scaffolds to display antagonistic peptides. AB - Neutralizing of TNF-alpha has been proved effective in treatment of some autoimmune diseases, e.g. rheumatoid arthritis and Crohn's disease. Low molecular weight synthetic peptides can mimic the binding sites of TNF-alpha receptors and block the activity of TNF-alpha. In order to stabilize the conformation, increase the affinity and bioactivity, in this study, heavy chain variable region of human antibody was used as a scaffold to simultaneously display three peptides, which were designed on the interaction between TNF-alpha and it's neutralizing monoclonal antibody. On the basis of the structural character and physical chemical property of the families of seven kinds of heavy chain variable regions (VH) in human antibodies, the fifth type of VH was screened as scaffold to display the antagonist peptide. Based on the computer-guided molecular design method, a novel domain antibody against TNF-alpha (named as ATD5) was designed as TNF-alpha antagonist. The theoretical study showed that ATD5 was more stable than displayed antagonist peptide. The binding activity with TNF-alpha was higher than free peptides. After expression and purification in Escherichia coli, ATD5 could bind directly with TNF-alpha and inhibit the binding of TNF-alpha to its two receptors, TNFR1 and TNFR2. ATD5 could also reduce the TNF-alpha-mediated cytotoxicity and inhibit TNF-alpha-mediated caspase activation on L929 cells in a dose dependent manner. The activity of ATD5 was significantly stronger than three peptides displayed by ATD5. This study provides a novel strategy for the development of new TNF-alpha inhibitors. This study demonstrates that it is possible to screen potential antagonists of TNF-alpha using in vitro analysis systems in combination with the computer-aided modeling method. PMID- 17125838 TI - Synthesis, X-ray structure and antimycobacterial activity of silver complexes with alpha-hydroxycarboxylic acids. AB - In this paper, synthesis, characterization and antimycobacterial properties of a new water-soluble complex identified as silver-mandelate are described. Elemental and thermal analyses are consistent with the formula [Ag(C(6)H(5)C(OH)COO)](n). The polymeric structure was determined by single X-ray diffraction and the two dimensional structure is based on the bis(carboxylate-O,O') dimer [Ag-O, 2.237(3), 2.222(3) Angstrom]. The structure is extended along both the b and c axes through two oxygen atoms of a bidentate alpha-hydroxyl-carboxylate residue [Ag-OH(hydroxyl), 2.477(3) Angstrom; Ag-O(carboxylate), 2.502(3) Angstrom; O-Ag O, 63.94(9) degrees]. A strong d(10)-d(10) interaction was observed between two silver atoms. The Ag - Ag distance is 2.8307(15) Angstrom. The NMR (13)C spectrum in D(2)O shows that coordination of the ligand to Ag(I) occurs through the carboxylate group in solution. Potentiometric titration shows that only species with a molar metal:ligand ratio of 2:2 are formed in aqueous solution. The mandelate complex and the silver-glycolate, silver-malate and silver-hydrogen tartarate complexes were tested against three types of mycobacteria, Mycobacterium avium, Mycobacterium tuberculosis and Mycobacterium kansasii, and their minimal inhibitory concentration (MIC) values were determined. The results show that the four complexes are potential candidates for antiseptic or disinfectant drugs for discharged secretions of patients affected with tuberculosis. PMID- 17125839 TI - Self-assembly and cytotoxicity study of waterwheel-like dinuclear metal complexes: the first metal complexes appended with multiple free hydroxamic acid groups. AB - Two waterwheel-like dinuclear complexes [M(2)(PHA)(4)(H(2)O)(2)] (M = Cu(II) (1), Zn(II) (2); HPHA = phthal-hydroxamic acid) appended with four free hydroxamic acid groups, namely, free hydroxamic acid metal complexes (FHAMCs) have been synthesized and characterized. The crystal structure of complex 1 was determined by single crystal X-ray diffraction, which adopts the paddlewheel motif with four bidentate carboxylate ligands joining two Cu(II) ions. The relative cytotoxicities of compounds 1 and 2 against SMMC-7721 and HO-8910 cell lines are similar and more predominant than HPHA (IC(50): Cu(II)>Zn(II)>>HPHA). The synergic effect of the bound water molecules, multiple free hydroxamic acid groups and dimetal active sites with bridging carboxylate may have significant impacts on their pharmacological activity. As the prototype for a new class of hydroxamic acid derivatives, the self-assembly of FHAMCs presents a promising new strategy in designing multiple hydroxamic acids with remarkable bioactivities. PMID- 17125840 TI - Aortic valve infective endocarditis caused by Serratia liquefaciens. AB - Infective endocarditis of the cardiac valves caused by Serratia liquefaciens has never been reported. We describe a fatal case of aortic valve infective endocarditis caused by S. liquefaciens after intravenous injection of amphetamine, complicated by multiple organ dysfunctions. PMID- 17125841 TI - Type 1 angiotensin receptor pharmacology: signaling beyond G proteins. AB - Drugs that inhibit the production of angiotensin II (AngII) or its access to the type 1 angiotensin receptor (AT(1)R) are prescribed to alleviate high blood pressure and its cardiovascular complications. Accordingly, much research has focused on the molecular pharmacology of AT(1)R activation and signaling. An emerging theme is that the AT(1)R generates G protein dependent as well as independent signals and that these transduction systems separately contribute to AT(1)R biology in health and disease. Regulatory molecules termed arrestins are central to this process as is the capacity of AT(1)R to crosstalk with other receptor systems, such as the widely studied transactivation of growth factor receptors. AT(1)R function can also be modulated by polymorphisms in the AGTR gene, which may significantly alter receptor expression and function; a capacity of the receptor to dimerize/oligomerize with altered pharmacology; and by the cellular environment in which the receptor resides. Together, these aspects of the AT(1)R "flavour" the response to angiotensin; they may also contribute to disease, determine the efficacy of current drugs and offer a unique opportunity to develop new therapeutics that antagonize only selective facets of AT(1)R function. PMID- 17125842 TI - Rapid high-yield mRNA extraction for reverse-transcription PCR. AB - Reverse-transcription PCR (RT-PCR) is the gold standard for mRNA quantification. Efficient, rapid, and high-throughput mRNA extraction is a prerequisite to ensure PCR sensitivity and precision, particularly for quantification of low-abundance mRNAs, and for large numbers of samples. Many mRNA extraction methods entail meticulous handling of individual samples, and are not well suited for large sample numbers. To achieve simple separation of mRNA binding matrix and the medium from which mRNA is to be isolated, oligo (dT)(20)-coated silica beads were used. Simple centrifugation and decanting steps can be used throughout the extraction procedure to separate supernatant fluids from the silica beads. DNase treatment reduced clumping of sedimented beads, thus facilitating bead resuspension and avoiding repeated agitation. DNase treatment also significantly reduced contaminating DNA, increased mRNA purity, and enhanced mRNA PCR readout by approximately 5-fold. The number of target transcripts per sample aliquot was higher in DNase-treated mRNA than in non-treated mRNA or in total nucleic acids. Thus, use of DNase-treated mRNA increased sensitivity of detection and quantification of low-copy transcripts. In conclusion, we describe here a simple, rapid, and cost-effective method that facilitates convenient extraction of high quality mRNA by minimizing cumbersome mechanical disruption and pipetting steps. PMID- 17125843 TI - What does Chilean research tell us about postpartum depression (PPD)? AB - BACKGROUND: In Chile, a country with a so called emerging market-economy, where rapid social and life style changes are taking place, women and the more socially disadvantaged are more at risk of becoming depressed. METHODS: Results of several studies are summarized in the context of a review of the literature. RESULTS: A third of Chilean women have depressive and/or anxiety symptoms during midpregnancy, while prevalence figures both in the early and the late postpartum period increase up to 50% in most studies. If strict operational criteria describing well defined depressive disorders are used postnatally, differences in prevalence and incidence figures arise depending on socioeconomic status. Whereas incidence rates for postpartum depression (around 9%) are very similar to those found in the northern hemisphere and do not appear to vary across different socioeconomic levels, higher prevalence rates are found among women from lower socioeconomic status. LIMITATIONS: The studies focused on current diagnostic entities and did not consider different clusters or dimensions. CONCLUSION: A shared biological etiology may be triggered by the physiology of childbirth and account for similarities in incidence across different socioeconomic levels. In turn, we hypothesize that the higher prevalence of postpartum depression (PPD) in Chilean women from lower socioeconomic status is the result of pre-existing depression and is not caused by more new cases of the illness. PMID- 17125844 TI - Factors influencing mother-child reports of depressive symptoms and agreement among clinically referred depressed youngsters in Hungary. AB - BACKGROUND: Psychiatric assessments of children typically involve two informants, the child and the parent. Understanding discordance in their reports has been of interest to clinicians and researchers. We examine differences between mothers' and children's report of children's depressive symptom severity, and factors that may influence their reports and level of agreement. We hypothesized that agreement between mother and child would improve if (1) the mother is depressed, due to improved recall of mood congruent symptoms, (2) the child is older, due to better social-cognitive and communication skills, and (3) the child is a female. METHODS: Subjects were 354 children (158 girls; mean age 11.69 years, SD: 2.05 years) with Major Depressive Disorder. Depressive symptoms were evaluated by a semi-structured interview separately with the mother and the child. Agreement on symptom severity was based on concordance of the presence and extent of symptoms. RESULTS: Maternal reports were significantly higher than their son's but not daughters'. Girls, particularly with increasing age, reported higher levels of symptoms; however mothers' reports were not affected by child sex or age. Maternal depression predicted more severe symptom reports for both children and mothers. Agreement between the mother and the child increased as children got older. LIMITATIONS: The same clinician interviewed the mother and the child, which might inflate rates of agreement. However, this method mirrors clinical evaluation. CONCLUSION: During a clinical interview one must consider the age and sex of the child and the depressive state of the mother in assimilating information about the child. PMID- 17125845 TI - Use of the process dissociation procedure to study the contextual effects on face recognition in schizophrenia: familiarity, associative recollection and discriminative recollection. AB - Contextual effects were explored in schizophrenia patients and paired comparison subjects during a long-term face recognition task. The objective was to investigate the contextual effects on face recognition by manipulating, in the same experiment, the perceptual context of the face (intrinsic vs. extrinsic) and the task context (inclusion vs. exclusion instructions). The situation was derived from the Jacoby's [Jacoby, L.L., 1991. A process dissociation framework: separating automatic from intentional uses of memory. Journal of Memory and Language 30, 513-541] process dissociation procedure. The results showed that schizophrenia patients (N=20) presented lower performances than healthy controls (N=20) in the inclusion but not in the exclusion task. This observation emphasizes the heterogeneity of recollection and suggests that the memory impairment in schizophrenia reflects an imbalance between two mechanisms. The first is a deficit in "associative recollection", i.e., the failure to use efficiently associative information. The other is an enhanced "discriminative recollection" that impedes their capacity to process information separately from its perceptual context. In addition, correlation with symptoms suggest that the former is expressed in the loosening of associations characteristic of disorganization symptoms, whereas the latter reflects the lack of flexibility or the contextualization bias related to psychotic symptoms, i.e., delusions and hallucinations. PMID- 17125846 TI - Matrix metalloproteinase mRNA expression in canine anal furunculosis lesions. AB - Although the aetiology of anal furunculosis (AF) in dogs is poorly understood, there is evidence for an underlying immune dysfunction. This is illustrated by the presence of a T helper type 1 cytokine mRNA profile in AF lesions and the clinical response to ciclosporin therapy. Expression of MMPs 2, 9 and 13 were evaluated in AF lesional biopsies by real-time quantitative RT-PCR. There was significantly increased expression of both MMP-9 and MMP-13 mRNA in AF biopsies compared to controls (p<0.001) but no significant difference in MMP-2 mRNA expression. Since MMP-9 and MMP-13 are primarily produced by macrophages, these data suggest that ulceration could be the result of aberrant activation of this cell type in the tissues. It is feasible that such pathological macrophage activity occurs in response to interferon-gamma secreted by T helper type 1 cells. This could explain why the lesions resolve following treatment with the immunosuppressive drug ciclosporin. PMID- 17125847 TI - Immunopathogenesis of adrenoleukodystrophy: current understanding. AB - Adrenoleukodystrophy is a neurometabolic disease with a decreased ability to degrade very long chain fatty acids (VLCFA) and significant phenotypic variation. Unlike most neurometabolic diseases, the success of hematopoietic stem cell transplantation (HSCT) is based on acquiring a new immune system rather than enzyme replacement. VLCFA accumulation appears necessary but not sufficient for pathogenesis. Evidence for the involvement of different components of the immune system in the pathogenesis of the cerebral lesions (cellular, cytokines, humoral, and complement) is reviewed, along with both HSCT and non-HSCT immunologic approaches to treatment and future directions. PMID- 17125848 TI - Melatonin is responsible for the nocturnal increase observed in serum and thymus of alpha1-thymosin and thymulin concentrations: observations in rats and humans. PMID- 17125849 TI - Late diagnosis of foreign body aspiration in children with chronic respiratory symptoms. AB - BACKGROUND: Children with undiagnosed and retained foreign bodies (FBs) may present with persistent respiratory symptoms. Delayed diagnosis is an important problem in developing countries and several factors affect the delay. OBJECTIVES: To investigate, the incidence of clinically unsuspected foreign body aspiration (FBA) in our flexible bronchoscopy procedures, the causes resulting in late diagnosis of FBA, and the incidence of the complications of FBA according to elapsed time between aspiration and diagnosis. METHODS: We reviewed the records of all the patients who underwent flexible bronchoscopy between 1997 and 2004 in our clinic. Patients with FBA were identified and their medical records were reviewed. RESULTS: During the study period, 654 children underwent flexible bronchoscopy; 32 cases (4.8%) of FBA were identified. Median age of patients was 29.5 months at presentation with a median symptomatic period of 3 months. None of the patients had a history of FBA. The most common misdiagnosis was bronchitis. Flexible bronchoscopy was performed to these patients within 1 week following presentation. In 87% of the patients (n=28), FBs were in organic nature. Patients were followed up for 21.0 months after removal of the FBs. Fifty-three percent (n=17) of the patients had a complete remission after bronchoscopic removal of the FBs. However, nine (28.8%) patients had chronic respiratory problems and six patients (18.8%) developed bronchiectasis. CONCLUSIONS: Atypical or prolonged respiratory symptoms should alert the physician and clinical and radiological findings should be carefully evaluated for a possible FBA. Delay in diagnosis and treatment of FBA should be avoided to prevent complications. PMID- 17125850 TI - Antiviral activity of tiazofurin and mycophenolic acid against Grapevine leafroll associated virus 3 in Vitis vinifera explants. AB - The ability to control plant viral diseases with chemicals has great potential value for agriculture, but few chemicals are available to date due to the difficulty in obtaining effective drugs. IMP dehydrogenase is an enzyme which catalyzes the conversion of inosine 5'-monophosphate to xanthosine 5' monophosphate in the de novo purine nucleotide synthetic pathway, and is considered a sensitive target for antiviral drugs. Two IMPDH inhibitors, tiazofurin (TR) and mycophenolic acid (MPA), were tested for their inhibitory effect on Grapevine leafroll-associated virus 3 (GLRaV-3) in in vitro grapevine explants. TR administration produced plantlets characterized by negative ELISA readings. No PCR products were obtained from these samples. This was confirmed by the absence of viral particles. MPA was essentially ineffective against GLRaV-3 replication in Sangiovese explants. This is the first report of GLRaV-3 eradication in grapevine explants following TR administration. PMID- 17125851 TI - Vulnerable windows for developmental ethanol toxicity in the Japanese medaka fish (Oryzias latipes). AB - Ethanol (EtOH) is a well-known developmental toxicant that produces a range of abnormal phenotypes in mammalian systems including craniofacial abnormalities, cognitive deficits and growth retardation. While the toxic potential of developmental EtOH exposure is well characterized clinically, the effect of timing on the extent of toxicity remains unknown. Fish models such as the Japanese medaka, Oryzias latipes, provide a convenient system for investigating the effects of developmental EtOH exposure in vivo. In this study, medaka embryo toxicity tests were used to assess temporal variations in developmental EtOH toxicity. Fertilized eggs were collected and incubated during early, middle or late egg development (e.g., 0-3, 3-6 or 6-9 days post-fertilization) with various sub-lethal concentrations of EtOH [0.1% (17.2 mM), 0.5% (86.0 mM) or 1% (172 mM)]. Uptake of EtOH by the embryo was 60-68% of the solution concentration across all windows. Time to hatch, head width, total body length and whole embryo caspase activity were used to assess toxicity. Hatching delays were noted only at the highest concentration of EtOH. Head width was affected at all ethanol levels, regardless of the window of exposure. EtOH-induced decreases in body length, however, appeared to be most pronounced when exposure occurred either during the first or last window. The effect on caspase-3/7 activity also depended on the window of exposure, with increases in caspase noted in embryos treated on days 1 or 2 (first window) and decreases seen in embryos treated on day 6 (second window) or day 8 (third window). In general, these data suggest that critical periods for heightened sensitivity to developmental EtOH exposure may vary according to the specific endpoint used to assess toxicity. PMID- 17125852 TI - Renal responses to acute lead waterborne exposure in the freshwater rainbow trout (Oncorhynchus mykiss). AB - The possible nephrotoxic effects of waterborne lead exposure (as Pb(NO3)2) were investigated in the freshwater rainbow trout (Oncorhynchus mykiss). Kidney lead accumulation was time-dependent, increasing upon exposure to 0.57+/-0.01 mg dissolved Pb L(-1) for up to 96 h with a significantly higher burden occurring in the posterior kidney compared to the anterior segment. Urine analyses in trout exposed to 1.20+/-0.09 mg dissolved Pb L(-1) revealed a significant increase in urinary lead excretion rate throughout 96 h of exposure. Urine flow rate and glomerular filtration rate (GFR) were not impacted with the exception of a significant decrease in GFR from 84 to 96 h in lead-exposed trout. Urine pH decreased significantly over time in lead-exposed fish. Correspondingly, urine ammonia excretion rate showed a marked increase from 48 h onwards. In experimental fish, urine glucose excretion was significantly greater by 96 h while urine lactate, urea and protein excretion were not significantly altered by lead exposure. The urine excretion rate of Ca2+ increased significantly by approximately 43% after only 24 h of lead exposure, and was maintained at a higher rate than controls for up to 96 h. Magnesium excretion increased in a time dependent fashion, reaching a two- to three-fold rise by 96 h. In contrast, rates of Na+ and Cl- excretion were decreased in experimental fish by approximately 30% by 48 h, this trend continuing for the duration of lead-exposure. There were no changes in any of these parameters in similarly treated control fish. Clearance ratio analyses indicated progressive decreases in the net reabsorption efficiencies of the renal system for Ca2+, Mg2+, Pb, and glucose, suggesting that the active tubular transport mechanisms for these substances were inhibited by lead exposure, while Na+, K+, Cl-, lactate, and protein reabsorptions were unaffected. Net ammonia secretion increased. We conclude that changes in renal function both reflect and help to minimize some of the associated disturbances in systemic physiology. Lead-induced ionoregulatory toxicity in rainbow trout, particularly the disturbance of Ca2+ homeostasis, is not exclusively a branchial phenomenon, but is in part a result of disruption of ionoregulatory mechanisms at the kidney. This action of lead outside the gills is critical to consider when developing guidelines for water quality. PMID- 17125853 TI - EhGEF2, a Dbl-RhoGEF from Entamoeba histolytica has atypical biochemical properties and participates in essential cellular processes. AB - Dbl proteins are a family of factors that exchange the guanine nucleotide which promote the activation of Rho small GTPases. This paper reports the molecular, structural, biochemical and functional characterization of EhGEF2, a new member of the Dbl family. EhGEF2 is the second GEF studied in parasites and in the protozoan Entamoeba histolytica, and it is also the first member of the Dbl family that was found to have Arm repeats. The catalytic domain (DH) of EhGEF2 has the conserved residues T421, N590 and E591, which are important for the activation of the GTPases. Biochemical studies on EhGEF2 showed that it could activate in vitro the amoebic GTPases EhRacA, EhRacB, EhRacC, EhRacD, EhRacG, EhRacH and EhCdc42, being EhRacG its main target. It was found that the DH domain binds specifically phosphatidic acid (PA); docking and lipid dot blot studies indicated that this binding does not interfere with the contact surface of EhRacG. Functional studies showed that both the Arm repeats and the catalytic domain of EhGEF2 participate in its localization at the amoebic membrane. Expression of a negative dominant version of EhGEF2 protein in E. histolytica provoked a 30% decrease in its ability to phagocyte human erythrocytes as well as severe effects on both the proliferation and the cellular chemotaxis which suggest that EhGEF2 participates in these cellular processes. PMID- 17125854 TI - Genome-scale protein expression and structural biology of Plasmodium falciparum and related Apicomplexan organisms. AB - Parasites from the protozoan phylum Apicomplexa are responsible for diseases, such as malaria, toxoplasmosis and cryptosporidiosis, all of which have significantly higher rates of mortality and morbidity in economically underdeveloped regions of the world. Advances in vaccine development and drug discovery are urgently needed to control these diseases and can be facilitated by production of purified recombinant proteins from Apicomplexan genomes and determination of their 3D structures. To date, both heterologous expression and crystallization of Apicomplexan proteins have seen only limited success. In an effort to explore the effectiveness of producing and crystallizing proteins on a genome-scale using a standardized methodology, over 400 distinct Plasmodium falciparum target genes were chosen representing different cellular classes, along with select orthologues from four other Plasmodium species as well as Cryptosporidium parvum and Toxoplasma gondii. From a total of 1008 genes from the seven genomes, 304 (30.2%) produced purified soluble proteins and 97 (9.6%) crystallized, culminating in 36 crystal structures. These results demonstrate that, contrary to previous findings, a standardized platform using Escherichia coli can be effective for genome-scale production and crystallography of Apicomplexan proteins. Predictably, orthologous proteins from different Apicomplexan genomes behaved differently in expression, purification and crystallization, although the overall success rates of Plasmodium orthologues do not differ significantly. Their differences were effectively exploited to elevate the overall productivity to levels comparable to the most successful ongoing structural genomics projects: 229 of the 468 target genes produced purified soluble protein from one or more organisms, with 80 and 32 of the purified targets, respectively, leading to crystals and ultimately structures from one or more orthologues. PMID- 17125855 TI - Subcortical vascular lesions and functional recovery in older patients with gait disorders. AB - This study aimed to assess whether subcortical vascular lesions (SVLs) predict functional recovery after rehabilitation in elderly patients with gait disorders (GD) due to multiple etiology (GD-ME). All patients consecutively admitted with GD-ME (n=103) in our Rehabilitation and Aged Care Unit (RACU) underwent a standardized rehabilitative program. The outcome measure was the Barthel Index (BI) Relative Functional Gain (RFG), a measure of improvement adjusting for baseline functional level. Potential predictors included cognition, depression, functional and nutritional status, physical health, occurrence of adverse clinical events during hospital stay, and SVLs, assessed with a validated visual rating scale based on brain CT scans. Predictors were divided into quartiles and the association with RFG was assessed. In a multivariate linear regression model, SVLs maintained its predictive power on RFG after adjustment for age, gender, and adverse clinical events, which was the only variable associated to RFG in the bivariate model (adjusted p=0.002 for trend). The study shows that SVLs is a predictor of functional recovery in elderly patients with GD-ME. PMID- 17125856 TI - Prompt resolution of an apical left ventricular thrombus in a patient with takotsubo cardiomyopathy. AB - This report describes the prompt resolution of an apical left ventricular (LV) thrombus complicating transient apical ballooning in a 74-year-old woman. The patient was admitted to our emergency department with acute chest pain and ST elevation on the electrocardiogram. Coronary angiography showed normal coronary arteries and LV-angiography demonstrated the presence of apical ballooning akinesis associated with basal hypercontraction. Echocardiography and MRI studies confirmed the presence of LV-apex akinesis and detected an apical thrombus. Follow-up echocardiography on day 12 before discharge of the patient, revealed a marked improvement of regional contractility of the LV-apex and surprisingly the complete resolution of the LV-apical thrombus. The patient was diagnosed with takotsubo cardiomyopathy. PMID- 17125857 TI - Polymorphic ventricular tachycardia induced by coronary vasospasm: a malignant case of variant angina. AB - Variant angina is generally a benign disease with self limiting symptoms. But in some cases serious ventricular arrhythmias which can lead to death can be seen. In this paper we present a case of variant angina who subsequently developed polymorphic ventricular tachycardia and was treated successfully with long acting nitrate and calcium channel blockade therapy. PMID- 17125858 TI - Biventricular involvement in a Turkish boy with palmoplantar hyperkeratosis and curly hair, an unusual presentation of Naxos-Carvajal syndrome. AB - Naxos disease is an autosomal recessively inherited familial syndrome characterized by woolly hair, palmoplantar keratoderma and a cell adhesion cardiomyopathy, especially arrhythmogenic right ventricular dysplasia (ARVD). Carvajal syndrome is a variant of Naxos disease in which curly or woolly hair, biventricular--predominantly left ventricular involvement were seen. Mutations in genes encoding the cell adhesion proteins like plakoglobin and desmoplakin were related with these syndromes. We report a 17-year-old boy and his family findings with curly hair, palmoplantar hyperkeratosis, ARVD and left ventricular involvement. The family was of Arabic origin, and a third-degree consanguinity was reported between the parents. They are from east part of Turkey and there were no relatives from Cyclades Island (Greece). Patient's younger brother had ARVD without cutaneous manifestations of the syndrome and his grand father had mild ARVD, curly hair and palmoplantar hyperkeratosis. PMID- 17125859 TI - Importance of the CT/MRI fusion method as a learning tool for CT-based postimplant dosimetry in prostate brachytherapy. AB - BACKGROUND AND PURPOSE: To compare the CT-based and CT/MRI fusion-based postimplant dosimetry after permanent prostate brachytherapy and to evaluate the improvement in CT-based dosimetry by physicians with or without experience in using the CT/MRI fusion method. PATIENTS AND METHODS: Thirty-eight consecutive patients agreed to participate in a prospective study. The prostate contours from CT/MRI fusion are the gold standard for determining the prostate volume and dose volume histogram (DVH). CT-based postimplant dosimetries were performed by two physicians. Observer 1 was a radiologist who had never used CT/MRI fusion method for postimplant dosimetric analysis. Observer 2 was a radiation oncologist experienced in postimplant analysis using the CT/MRI fusion method. The prostate dosimetry was evaluated by prostate D90 and V100. RESULTS: No significant difference was observed in the mean prostate volumes between the two observers and the CT/MRI fusion data. However, the correlation coefficient value for observer 2 (R(2)=0.932) was greater than that for observer 1 (R(2)=0.793). The D90 and V100 values as evaluated by the two observers were significantly underestimated in comparison to those evaluated using the CT/MRI fusion methods. The DVH related parameters were underestimated more frequently by observer 1 than by observer 2: (prostate D90: 99.56% for observer 1, 102.97% for observer 2, 109.37% for CT/MRI fusion. Prostate V100: 88.12% for observer 1, 90.14% for observer 2, 91.91% for CT/MRI fusion). CONCLUSIONS: The difference in the mean value in D90 and V100 by observer 1 was significantly greater than that for observer 2. These findings suggest that the CT/MRI fusion method provides accurate feedback which thereby improves CT-based postimplant dosimetry for prostate brachytherapy. PMID- 17125860 TI - Prostate displacement kit: reducing the radiation dose to the rectum. AB - During high dose-rate brachytherapy boost in 20 patients the use of a prostate water-rectal-displacement-kit contributed to an increase in the distance between the prostate and the rectum, however, the prostate was not totally immobilized by the needles, implying the necessity for an very careful on-line dose-planning dosimetry. PMID- 17125861 TI - Involved-node radiotherapy to the mediastinum. PMID- 17125862 TI - Dosimetric verification of the anisotropic analytical algorithm for radiotherapy treatment planning. AB - BACKGROUND AND PURPOSE: To investigate the accuracy of photon dose calculations performed by the Anisotropic Analytical Algorithm, in homogeneous and inhomogeneous media and in simulated treatment plans. MATERIALS AND METHODS: Predicted dose distributions were compared with ionisation chamber and film measurements for a series of increasingly complex situations. Initially, simple and complex fields in a homogeneous medium were studied. The effect of inhomogeneities was investigated using a range of phantoms constructed of water, bone and lung substitute materials. Simulated treatment plans were then produced using a semi-anthropomorphic phantom and the delivered doses compared to the doses predicted by the Anisotropic Analytical Algorithm. RESULTS: In a homogeneous medium, agreement was found to be within 2% dose or 2mm dta in most instances. In the presence of heterogeneities, agreement was generally to within 2.5%. The simulated treatment plan measurements agreed to within 2.5% or 2mm. CONCLUSIONS: The accuracy of the algorithm was found to be satisfactory at 6 and 10MV both in homogeneous and inhomogeneous situations and in the simulated treatment plans. The algorithm was more accurate than the Pencil Beam Convolution model, particularly in the presence of low density heterogeneities. PMID- 17125863 TI - Opening lecture of the 25th annual meeting, ESTRO, Leipzig expensive new therapy and finite resources: an ethical dilemma for doctors and scientists. PMID- 17125865 TI - High-dose brachytherapy using MammoSite applicator: "Our doubts are traitors, and makes us lose the good we oft might win, by fearing to attempt !". PMID- 17125866 TI - Influence of the initial rectal distension on posterior margins in primary and postoperative radiotherapy for prostate cancer. AB - BACKGROUND AND PURPOSE: The aim of the study was to define the effect of different rectum fillings in the planning CT study on the posterior clinical target volume (CTV) displacements (PD) in primary and postoperative radiotherapy (RT) for prostate cancer. MATERIALS AND METHODS: Fifty patients underwent CT scans in supine position with a full bladder and an empty bladder before RT and at several points in time during the treatment. PD were determined depending on the initial rectum volume (RV), average cross-sectional rectal area (CSA), and the rectal diameter at the level of the bladder neck (RD). RESULTS: Posterior CTV motion was not found to be minimal with a particularly small initial rectum filling. Steeply increasing PD resulted for patients with RV>120cm(3), CSA>12cm(2), and RD>4.5cm. While below these critical values a posterior margin of 6mm/9mm allowed to cover 80%/90% of displacements, 18mm/24mm were needed for patients with larger rectum fillings. No correlation of increasing rectum distension with increasing PD was found at the apex level. PD could not be reduced by voiding the bladder. CONCLUSIONS: Defining the posterior margin in prostate RT, the initial rectum distension and the superior-inferior CTV level has to be considered. Patients with large initial rectum fillings have preferentially the need for repeated planning CT scans or image-guided RT. PMID- 17125867 TI - N-Acetylcysteine and ebselen but not nifedipine protected cerebellar granule neurons against 4-hydroxynonenal-induced neuronal death. AB - 4-Hydroxynonenal (HNE), an aldehydic product of membrane lipid peroxidation, has been shown to induce neurotoxicity in various types of neurons. To clarify the mechanisms underlying HNE-induced neurotoxicity, the effects of antioxidants (N acetylcysteine (NAC) and ebselen with or without NAC pretreatment) and Ca(2+) related reagents were examined in cerebellar granule neurons. The decreases in neuronal survival and mitochondrial membrane potential induced by HNE were suppressed by pretreatment with NAC at concentrations of 500 and 1000 microM. HNE induced protein modification and reactive oxygen species generation were also suppressed by pretreatment with NAC at 1000 microM. Although simultaneous application of ebselen (10 microM) did not protect against HNE-induced neurotoxicity, it completely suppressed HNE-induced injury after pretreatment with NAC at 300 microM. HNE increased [Ca(2+)](i) levels, and this increase was significantly attenuated by simultaneous application of nifedipine (10 microM) or EGTA (1000 microM), but not by MK-801 or CNQX. However, none of these Ca(2+) related reagents was able to prevent HNE-induced neuronal death or mitochondrial injury. These results suggest that pretreatment with a low concentration of NAC dramatically potentiates the neuroprotective activity of ebselen, and that HNE induced increase in [Ca(2+)](i) is not involved in HNE-induced neuronal death in cerebellar granule neurons. PMID- 17125868 TI - Altered levels of serotonin in lymphoblasts derived from migraine patients. AB - We previously reported that dysfunctions in the autonomic nervous systems of patients with migraines occur not only in the brain, but throughout the whole body. Serotonin and neuropeptides are also known to have important roles in the pathophysiology of migraine. With this background in mind, we analyzed human lymphoblast cell lines from migraine with aura (MwA) patients to investigate the pathophysiology of migraine. The characteristics of these lymphoblasts and the involvement of the lymphoblasts in serotonin metabolism were examined. The lymphoblasts expressed serotonin receptors as well as some enzymes related to serotonin metabolism. The serotonin level in the MwA lymphoblasts was higher than that in the control cells. However, serotonin uptake into the lymphoblasts in MwA patients was similar to that in the control subjects. These findings suggest that lymphoblasts in MwA patients have altered levels of serotonin metabolism. Moreover, we propose that this lymphoblast cell system could serve as a novel modality for migraine research. PMID- 17125869 TI - Risk evaluation for staphylococcal food poisoning in processed milk produced with skim milk powder. AB - The growth of S. aureus and the production of staphylococcal enterotoxin A (SEA) in skim milk concentrates stored at inappropriate temperatures in a recovery milk tank (tank for excess concentrated skim milk) used in the manufacture of skimmed milk powder were investigated. Also, it was estimated if a possible outbreak of food poisoning would occur if the contaminated skimmed milk powder was used in the manufacture of processed milk. Skim milk concentrates with milk solid content of 15, 25, and 35% were inoculated with S. aureus at 1-2 log CFU/ml and incubated at 15, 25, or 35 degrees C for 0 to 24 h with or without shaking. Bacterial growth and the level of SEA production were measured. At 35 degrees C with shaking, there was a significant difference (p<0.05) in one way layout analysis of variance, and it was demonstrated that the growth of S. aureus and SEA production could be milk solid content-dependent. Shaking accelerated the growth of S. aureus and SEA production at 35 degrees C. Generally, skim milk powder is produced by mixing a set percentage of skim milk concentrates (recovery milk) from the recovery milk tank into raw milk. If recovery milk contaminated with S. aureus at levels of 1-2 log CFU/ml is kept at 15 to 35 degrees C due to a power failure, it was estimated that processed milk consumption of 670-1200 ml, 420 1500 ml and 18-83 ml would trigger the onset of food poisoning symptoms when skim milk concentrates (recovery milk) are stored at 25 degrees C for 24 h, 35 degrees C for 10 h, and 35 degrees C for 24 h, respectively, during the production of the skim milk powder. Based on these consumption levels, it was concluded that, if recovery milk cannot be refrigerated and is stored at room temperature (25 to 35 degrees C), it must be used within 8 h and preferably within 6 h. PMID- 17125870 TI - Transcript stabilization by mRNA sequences from hrpA of Pseudomonas syringae. AB - Production of heterologous proteins in bacteria is one of the main applications of biotechnology. Although several high-efficiency expression systems have been developed, different steps in protein production may become rate-limiting depending on the production system and the protein being produced. One bottle neck can be the instability of the mRNA. We have used fragments of the unusually long-living mRNA hrpA from the plant pathogenic bacteria Pseudomonas syringae pathovars tomato and phaseolicola to increase the half-lives of heterologous transcripts. The stabilizing effect was extended to Escherichia coli, as half lives of several heterologous transcripts were increased from a few minutes to up to 19min. Production of heterologous proteins was also increased manifold by the addition of the stabilizing hrpA elements. We have mapped the regions of the hrpA transcript necessary and sufficient for the stabilization process. PMID- 17125871 TI - A single stem-loop structure in Tacaribe arenavirus intergenic region is essential for transcription termination but is not required for a correct initiation of transcription and replication. AB - The genome of Tacaribe virus (TV), prototype of the New World arenaviruses, comprises two RNA segments each encoding two proteins in an ambisense orientation separated by an intergenic region (IGR). We used a TV minireplicon system to investigate the nature of the IGR structures required for transcription termination. We show that efficient generation of subgenomic (SG) RNAs is related to a single hairpin structure comprising a stem with variable numbers of uninterrupted base pairs and stabilized by high DeltaG values. The low ability of highly stable hairpin structures comprising bulged stems to support SG RNA synthesis suggested the importance of hairpin configuration for transcription termination. Neither the sequences downstream nor those upstream from the hairpin played a role in SG RNA accumulation. We also show that independently of the IGR structure the unencapsidated mRNAs contained short stretches of nontemplated bases at their 5' ends which are capped, whereas the 5' ends of the nucleocapsid associated antiminigenomes contained an uncapped extra residue. The results support the conclusions that: (i) transcription termination in TV is related to a structural element that is independent of sequence and (ii) the transcription termination signal is not required for a correct initiation of transcription and replication. PMID- 17125872 TI - Novel mutations found in mitochondrial diabetes in Chinese Han population. AB - Mitochondria provide cells with most of the energy in the form of ATP. Mutations in mitochondrial DNA (mtDNA) are associated with type 2 diabetes mellitus (T2DM) because ATP plays a critical role in the production and the release of insulin. To systematically determine mutant loci and to investigate their association with T2DM in Chinese Han population, 17 commonly reported mutant loci were screened in 236 cases of T2DM and 240 normal controls by PCR-RFLP, allele-specific PCR (AS PCR) and DNA sequencing methods. Biological softwares were used to analyze the secondary structure of DNA, RNA and the corresponding proteins for missense mutations. Sixteen mutant loci were detected in total, of which five were novel, GenBank accession nos. were DQ092356, DQ473644 and DQ473645; they were mainly in16S rRNA, ND1 and ND4 gene. There was significant difference between the two groups for ND1 and ND4 genes mutation frequencies (ND1: P=0.001, OR=3.944, 95% CI 1.671-9.306; ND4: P=0.010, OR=5.818, 95% CI 1.275-26.537). No significant association was observed between the two groups for 5178A/C polymorphisms (P=0.418). Our study suggested that T3394C and A12026G might be associated with T2DM in Chinese Han population, and T2DM with mtDNA variant should be considered mitochondrial diabetes. PMID- 17125873 TI - Prevention of severe toxic liver injury and oxidative stress in MCP-1-deficient mice. AB - BACKGROUND/AIMS: Administration of carbon tetrachloride determines liver injury, inflammation and oxidative stress, but the molecular mechanisms of damage are only partially understood. In this study, we investigated the development of acute toxic damage in mice lacking monocyte chemoattractant protein-1 (MCP-1), a chemokine which recruits monocytes and activated lymphocytes. METHODS: Mice with targeted deletion of the MCP-1 gene and wild type controls were administered a single intragastric dose of carbon tetrachloride. Serum liver enzymes, histology, expression of different chemokines and cytokines, and intrahepatic levels of oxidative stress-related products were evaluated. RESULTS: Compared to wild type mice, peak aminotransferase levels were significantly lower in MCP-1-deficient animals. This was paralleled by a delayed appearance of necrosis at histology. In addition, MCP-1-deficient mice showed a shift in the pattern of infiltrating inflammatory cells, with a predominance of polymorphonuclear leukocytes. Lack of MCP-1 was also accompanied by reduced intrahepatic expression of cytokines regulating inflammation and tissue repair. The increase in tissue levels of reactive oxygen species and 4-hydroxy-nonenal following administration of the hepatotoxin was also significantly lower in animals lacking MCP-1. CONCLUSIONS: Lack of MCP-1 affords protection from damage and development of oxidative stress in a toxic model of severe acute liver injury. PMID- 17125874 TI - Marked changes of the hepatic sinusoid in a transgenic mouse model of acute immune-mediated hepatitis. AB - BACKGROUND/AIMS: The liver sinusoidal endothelial cell (LSEC) is increasingly recognized as having an important role in hepatic immunity. However, the responses of LSECs and the hepatic sinusoid in immune-mediated hepatitis are poorly described. METHODS: We studied a transgenic mouse model of acute immune mediated hepatitis: Met-Kb mice injected with T cells from Des-TCR mice. RESULTS: Hepatitis was characterized by lymphocyte infiltrates causing severe but transient liver damage. There were marked changes in the ultrastructure of the LSEC five days after injection of the T cells that coincided with the peak of the hepatitis. The porosity of fenestrations in the LSEC decreased and the endothelium became thickened. LSECs appeared to be markedly activated. These changes were associated with narrowing of the space of Disse, loss of hepatocellular microvilli and deposition of basal lamina. Lymphocytes were seen passing through fenestrations. Loss of fenestration in the LSEC prevented hepatitis induced by a second injection of lymphocytes on day 5. CONCLUSIONS: Structural changes in the LSEC occur during the peak of a mouse model of immune mediated hepatitis. These changes were associated with attenuation of subsequent liver damage, suggesting that they may influence immunological responses mediated by LSECs or the passage of lymphocytes through LSEC fenestrations. PMID- 17125875 TI - IFN-gamma abrogates profibrogenic TGF-beta signaling in liver by targeting expression of inhibitory and receptor Smads. AB - BACKGROUND/AIMS: In a randomized open-labeled multicenter trial with patients suffering from chronic HBV infection, we recently identified a benefit of 9-month IFN-gamma treatment resulting in decreased fibrosis scores and a reduced number of alpha-smooth muscle actin-positive hepatic stellate cells (HSCs). Approaches opposing profibrogenic activities of TGF-beta may be amenable in chronic liver disease. According to experimental models, IFN-gamma counteracts several TGF-beta effects. METHODS: The crosstalk of IFN-gamma and TGF-beta signaling relevant for fibrogenesis was investigated in primary cultured rat HSCs and a cell line representing activated HSCs. RESULTS: In vitro studies with HSCs demonstrate that TGF-beta-dependent activation of (CAGA)9-MLP-Luc, a Smad3/4 responsive reporter construct, was significantly decreased by IFN-gamma, indicating a TGF-beta antagonizing function. IFN-gamma induced the activity of the Smad7 promoter and Smad7 protein expression via STAT-1 signaling. In contrast to TGF-beta, IFN-gamma was able to induce Smad7 expression in activated HSCs providing increased protein levels for at least 12h. In addition, expression of Smad2/3 was reduced by IFN gamma and activation of Smads2/3 was abrogated. CONCLUSIONS: IFN-gamma displays antifibrotic effects in liver cells via STAT-1 phosphorylation, upregulation of Smad7 expression and impaired TGF-beta signaling. PMID- 17125876 TI - Peginterferon alfa-2b and ribavirin in patients with hepatitis C virus and decompensated cirrhosis: a controlled study. AB - BACKGROUND/AIM: To evaluate long-term outcomes in decompensated HCV-related cirrhotic patients treated with antiviral therapy. METHODS: Of 129 eligible patients, 66 received peginterferon alfa-2b and ribavirin for 24 weeks, and 63 were controls. Survival and recurrence of liver failure events after therapy were main outcomes. RESULTS: Therapy was tolerated by 27 patients, dose reduced in 26 for toxicity, and discontinued in 13 for intolerance. End-of-therapy and sustained virological response (SVR) rates were 82.6% and 43.5% for HCV 2/3 patients, and 30.2% and 7.0% for HCV 1/4 patients. During therapy, odds ratios for severe infections or deaths due to infection were 2.95 (95% C.I. 0.93-9.3) and 1.97 (95% C.I. 0.40-9.51) in treated patients as compared with controls. During a follow-up of 30 months off-therapy, decompensated events occurred in 52, 33, and 3 of controls, non-responders, and SVR patients. Odds ratios for ascites, encephalopathy, and oesophageal bleeding in treated patients significantly decreased as compared with controls. Annualized incidence of death was 2.34, 1.91, and 0 per 1000 patient-years, respectively, in controls, non-responders, and SVR patients. Survival curves showed early separation of SVR patients from both non-responders and controls at approximately 6 months. CONCLUSIONS: In decompensated cirrhotics, HCV clearance by therapy is life-saving and reduces disease progression. PMID- 17125877 TI - Reduced susceptibility to epinephrine-induced arrhythmias in cirrhotic rats: the roles of nitric oxide and endogenous opioid peptides. AB - BACKGROUND/AIMS: The clinical relevance of QT prolongation, the most widely recognized cardiac electrophysiological abnormality of cirrhosis, is still undefined. The aim of this study is to examine the susceptibility of chronic (4 week) bile duct-ligated rats to epinephrine-induced arrhythmias. The roles of nitric oxide and endogenous opioids were also evaluated. METHODS: Sham-operated and cirrhotic rats were treated with daily subcutaneous administrations of normal saline (1 ml/kg/day), L-NAME (a non-selective nitric oxide synthase inhibitor, 3mg/kg/day), and naltrexone (20mg/kg/day) during the fourth week after operation. In order to evaluate the effects of acute nitric oxide synthesis inhibition, additional groups of animals were treated by acute intraperitoneal L-NAME injections (3mg/kg). Arrhythmias were induced by intravenous injections of 10 microg/kg epinephrine. RESULTS: Despite QT prolongation (P<0.001), epinephrine induced fewer arrhythmias in cirrhotic rats compared to sham-operated animals (P<0.05). Chronic, but not acute, L-NAME administration corrected the QT prolongation in cirrhotic rats (P<0.001), and restored the susceptibility of cirrhotic rats to arrhythmias (P<0.05). Naltrexone injection without a significant effect on epinephrine-induced arrhythmias corrected QT interval in cirrhotic rats (P<0.001). CONCLUSIONS: This study shows that despite QT prolongation, cirrhotic animals are resistant against epinephrine-induced arrhythmias. This resistance is mediated by chronic nitric oxide overproduction. PMID- 17125878 TI - Hepatitis E virus (HEV) infection in patients with cirrhosis is associated with rapid decompensation and death. AB - BACKGROUND/AIMS: India is hyper-endemic for hepatitis E virus (HEV). HEV infection in cirrhosis may cause high mortality. Prospective study evaluating HEV infection in cirrhotics is scarce. METHODS: Consecutive patients with cirrhosis and healthy controls were included. Cirrhotics were categorized to 3 groups, (Group I - rapid decompensation, Group II - chronically decompensated, Group III cirrhotics without decompensation). Sera from cirrhotics and controls were tested for HEV-RNA (RT-PCR). HEV-RNA positivity among cirrhotics and controls was compared. Natural course and mortality rate between HEV infected and non-infected cirrhotics were assessed during a 12-month follow-up. RESULTS: 107 cirrhotics and 200 controls were included. 30 (28%) cirrhotics and 9 (4.5%) controls had detectable HEV-RNA (p<0.001). HEV- RNA positivity among Group I (n=42), II (n=32) and III (n=33) cirrhotics was 21 (50%), 6 (19%) and 3 (10%), respectively (p=0.002). 70% (21/30) with HEV infection and 27% (21/77) without it had rapid decompensation (p=0.001). Mortality between HEV infected and non-infected cirrhotics at 4 weeks (43% vs. 22%, p=0.001) and 12 month (70% vs. 30%, p=0.001) was different. Multivariate analysis identified HEV infection, Child-Pugh's score, renal failure, and sepsis as independent factors for mortality. CONCLUSIONS: In India, cirrhotics were prone to HEV infection, which was associated with rapid decompensation and death. PMID- 17125879 TI - Polymorphisms of type I interferon receptor 1 promoter and their effects on chronic hepatitis B virus infection. AB - BACKGROUND/AIMS: Exposure to HBV leads to a distinct clinical course which is partially pertained to host genetic variability. We aimed to study polymorphisms of type I interferon receptor 1 (IFNAR1) promoter and their potential effects on chronic HBV infection. METHODS: Polymorphisms of IFNAR1 promoter were identified in 320 chronic hepatitis B patients, 148 spontaneously recovered individuals, 148 healthy Chinese donors and 114 Caucasians. Their functional capability in driving reporter gene expression was analyzed. RESULTS: Four polymorphic alleles were identified at loci -568, -408, -77 and -3. Association analysis revealed that carriers of alleles -568G, -408C and their related haplotype I were less susceptible to chronic HBV infection whereas those of alleles -568C, -408T and related haplotype III were significantly associated with higher risk to chronic hepatitis B (P<0.01). In a reporter-driven system, the promoter variants with alleles -408C and -3C could drive higher expression of the reporter gene than those with alleles -408T and -3T (P<0.01). Interestingly, an allele with 9 GT repeats at -77 that was rarely found in Chinese but prevalent in Caucasian exhibited the highest transcriptional ability. CONCLUSIONS: Our results showed that polymorphisms of IFNAR1 promoter may affect, at least in part, the outcomes of HBV infection. PMID- 17125880 TI - Remodelling of calcium signalling during liver regeneration in the rat. AB - BACKGROUND/AIMS: During liver regeneration, a network of cytokines and growth factors interact with hepatocytes, helping to restore the liver mass and functions after partial tissue loss. Agonists that trigger Ca2+ signals in the liver contribute to this process, although little is known about calcium signalling during liver regeneration. RESULTS: We observed two phases in which the hepatocyte response to calcium-mobilising agonists was greatly reduced versus control cells at 24h and five days after partial hepatectomy. We found that both phases of hepatocyte desensitisation involved the down-regulation of cell surface receptors and the type II InsP3 receptor. Single cell studies with flash photolysis of caged InsP3 revealed that InsP3-mediated Ca2+ release was slower in regenerating hepatocytes at 24, 48 h and 5 days than in control cells. Also, the temporal pattern of vasopressin-elicited intracellular calcium oscillations studied on fura2-loaded cells was altered, with the duration of each Ca2+ peak being longer. Finally, we showed an association between hepatocyte desensitisation and progression through the cell cycle towards the S phase at 24 h after hepatectomy. CONCLUSIONS: Our study supports the remodelling of hepatocyte calcium signalling during liver regeneration, and that this change is partly linked with cell cycle progression. PMID- 17125881 TI - Pricing and reimbursement of in-patent drugs in seven European countries: a comparative analysis. AB - The main objective of this comparative analysis was to assess regulations applied by EU governments to reward potentially innovative drugs. We focused on the pharmaceutical policy for in-patent drugs in seven EU countries: Belgium, France, Germany, Italy, the Netherlands, Spain, and the UK. A common scheme was applied to all seven countries: first, pricing and reimbursement procedures for new and innovative drugs were investigated; secondly, we focused on the use in the regulatory process of economic evaluations. The analysis involved reviewing the literature and interviewing a selected panel of local experts in each country. According to our comparative analysis, a first sensible step might be to classify active ingredients as those addressing neglected pathologies and those for diseases that are already successfully treated, thus offering more limited therapeutic gains by definition. A reasonable solution to reward real innovation could be to admit a premium price for very innovative drugs according to their estimated cost-effectiveness. New drugs with modest improvement could be grouped in therapeutic clusters and submitted to a common reference price, despite patent expiration. Such a "dual approach" could be a sensible compromise to restrict pharmaceutical expenditure while at the same time rewarding companies that invest in high-risk basic research. PMID- 17125882 TI - Which came first: insecticides or resistance? AB - Mutations that confer resistance to insecticides are well documented. However, so far, we have been unable to determine whether these mutations arose before or after the introduction of insecticides. Recently, a landmark study showed that resistance to Malathion can be detected in pinned specimens of Australian sheep blowflies that were collected before the introduction of the insecticide. This finding has numerous implications for our understanding of the prevalence of resistance to new compounds. It also indicates that pre-existing resistance alleles might not carry the fitness cost that is associated with new mutations. PMID- 17125883 TI - Synthetic approaches to uniform polymers. AB - Uniform polymers are characterised by a narrow molecular weight distribution (MWD). Uniformity is also defined by chemical structure in respect of (1) monomer orientation, sequence and stereo-regularity, (2) polymer shape and morphology and (3) chemical functionality. The function of natural polymers such as polypeptides and polynucleotides is related to their conformational structure (e.g. folded tertiary structure). This is only possible because of their high degree of uniformity. While completely uniform synthetic polymers are rare, polymers with broad structure and MWD are widely used in medicine and the biomedical sciences. They are integral components in final dosage forms, drug delivery systems (DDS) and in implantable devices. Increasingly uniform polymers are being used to develop more complex medicines (e.g. delivery of biopharmaceuticals, enhanced formulations or DDS's for existing actives). In addition to the function imparted by any new polymer it will be required to meet stringent specifications in terms of cost containment, scalability, biocompatibility and performance. Synthetic polymers with therapeutic activity are also being developed to exploit their polyvalent properties, which is not possible with low molecular weight molecules. There is need to utilise uniform polymers for applications where the polymer may interact with the systemic circulation, tissues or cellular environment. There are also potential applications (e.g. stimuli responsive coatings) where uniform polymers may be used for their more defined property profile. While it is not yet practical to prepare synthetic polymers to the same high degree of uniformity as proteins, nature also effectively utilises many polymers with lower degrees of uniformity (e.g. polysaccharides, poly(amino acids), polyhydroxyalkanoates). In recent years it has become possible to prepare with practical experimental protocols sufficient quantities of polymers that display many aspects of uniformity. This review describes practical strategies for polymer synthesis focusing on addition processes that have been developed that allow narrow MWD polymers to be prepared. Also described are some examples where aspects of polymer uniformity in terms of molecular weight and/or chemical constitution are exploited for their unique properties. PMID- 17125884 TI - Thermo- and pH-responsive polymers in drug delivery. AB - Stimuli-responsive polymers show a sharp change in properties upon a small or modest change in environmental condition, e.g. temperature, light, salt concentration or pH. This behaviour can be utilised for the preparation of so called 'smart' drug delivery systems, which mimic biological response behaviour to a certain extent. The possible environmental conditions to use for this purpose are limited due to the biomedical setting of drug delivery as application. Different organs, tissues and cellular compartments may have large differences in pH, which makes the pH a suitable stimulus. Therefore the majority of examples, discussed in this paper, deal with pH-responsive drug delivery system. Thermo-responsive polymer is also covered to a large extent, as well as double-responsive system. The physico-chemical behaviour underlying the phase transition will be discussed in brief. Then selected examples of applications are described. PMID- 17125885 TI - Attenuation of polycyclic aromatic hydrocarbons from urban stormwater runoff by wood filters. AB - A significant amount of contamination enters water bodies via stormwater runoff and, to reduce the amount of pollution, retention ponds are installed at many locations. While effective for treating suspended solids, retention ponds do not effectively remove dissolved constituents, such as polycyclic aromatic hydrocarbons (PAH). Previous laboratory studies demonstrates that aspen wood cuttings can be utilized to enhance the removal of dissolved contaminants. The objective of this pilot-scale field test was to determine if wood filters could effectively remove dissolved PAH from the runoff under field conditions. Four wood filter tests were conducted, lasting from 1 to 9 weeks, to determine the degree of PAH attenuation from the aqueous phase as a function of wood mass, residence times, and seasonable changes. The prototype wood filters removed on average between 18.5% and 35.6% (up to 66.5%) of the dissolved PAH contaminants. The PAH removal effectiveness of the wood was not affected by changes in water temperature or pH. The filter effectiveness increased with filter size and was highest in continuously submerged parts of the filter system. Also, heavier molecular weight PAH compounds (e.g. chrysene) were more effectively removed than lighter molecular weight compounds. Disassociation of weakly particle-bound PAH from the filter was identified as the most likely cause for a temporary drop of the wood filter's PAH load during intense storms. Simple filter design changes are likely to double the filter effectiveness and alleviate the disassociation problem. PMID- 17125887 TI - S-adenosylhomocysteine and polyunsaturated fatty acid metabolism in predementia syndromes and Alzheimer's disease. AB - Recent epidemiological evidence showed that dietary fatty acids, antioxidants, and micronutrients appear to have a role in cognitive decline, and may permit a beneficial effect on the risk of dementia and predementia syndromes. We discussed in the present paper the issue of the suggested protective role of polyunsaturated fatty acids (PUFA) on Alzheimer's disease, mild cognitive impairment, and other predementia syndromes, supported by the findings of the Italian Longitudinal Study on Aging (ILSA) and other population-based studies. In particular, we discussed the possible metabolic link of plasma S adenosylhomocysteine concentrations with PUFA erythrocyte composition in explaining this suggested protective role of fatty acids against dementia and predementia syndromes. PMID- 17125888 TI - Synthesis and antimicrobial activity of 1-(4-aryl-2-thiazolyl)-3-(2-thienyl)-5 aryl-2-pyrazoline derivatives. AB - Several 1-(4-aryl-2-thiazolyl)-3-(2-thienyl)-5-aryl-2-pyrazoline derivatives were synthesized by reacting substituted 3-(2-thienyl)-5-aryl-1-thiocarbamoyl-2 pyrazolines with phenacyl bromides in ethanol. The structures of the synthesized compounds were confirmed by (1)H NMR, (13)C NMR, and EIMS spectral data. Their antimicrobial activities against Escherichia coli (NRRL B-3704), Staphylococcus aureus (NRLL B-767), Salmonella typhimurium (NRRL B-4420), Bacillus cereus (NRRL B-3711), Streptococcus faecalis (NRRL B-14617), Aeromonas hydrophila (Ankara Uni. Fac. of Veterinary), Candida albicans and Candida glabrata (isolates obtained from Osmangazi Uni. Fac. of Medicine) were investigated. A significant level of activity was observed. PMID- 17125886 TI - Arrestins and two receptor kinases are upregulated in Parkinson's disease with dementia. AB - Arrestins and G proteins-coupled receptor kinases (GRKs) regulate signaling and trafficking of G protein-coupled receptors. We investigated changes in the expression of arrestins and GRKs in the striatum of patients with Parkinson's disease without (PD) or with dementia (PDD) at postmortem using Western blotting and ribonuclease protection assay. Both PD and PDD groups had similar degree of dopamine depletion in all striatal regions. Arrestin proteins and mRNAs were increased in the PDD group throughout striatum. Protein and mRNA of GRK5, the major subtype in the human striatum, and GRK3 were also upregulated, whereas GRK2 and 6 were mostly unchanged. The PD group had lower concentration of arrestins and GRKs than the PDD group. There was no statistical link between the load of Alzheimer's pathology and the expression of these signaling proteins. Upregulation of arrestins and GRK in PDD may confer resistance to the therapeutic effects of levodopa often observed in these patients. In addition, increased arrestin and GRK concentrations may lead to dementia via perturbation of multiple signaling mechanisms. PMID- 17125889 TI - [Recurrent delayed pericarditis after pacemaker implantation: a post pericardiotomy-like syndrome?]. AB - INTRODUCTION: Pacemaker implantation is a usual technique in cardiology which may be followed by acute pleural effusion and delayed unusual pericarditis. CASE REPORT: We reported the case of a 67 year-old man hospitalized for faintness. Rhythmical auricular disease was diagnosed and pacemaker was implanted without immediate complication. Though pericarditis with tamponade at the day 21 will require emergency pericardiotomy surgery. A recurrent pericarditis at day 45 was treated with anti-inflammatory drugs without relapse at the end of the treatment. DISCUSSION: Repeated delayed pericarditis after pacemaker surgery may be compared to the Dressler syndrome which occurs after myocardial infarction. PMID- 17125890 TI - Nursing education in Iran: past, present, and future. AB - Nursing education in Iran has undergone significant change since its genesis with foreign missionaries over one hundred years ago. More recently, following the 1979 Islamic revolution, nurse education has followed the direction taken by most other countries in moving from an apprenticeship model of training to an academic model. A series of transformative changes to nursing education specifically-and across the higher education system generally-has resulted in nurses now being able to undertake study across all university-based programs up to and including doctoral level. Contemporary nursing students have access to full-text professional journals through the internet, and they may pursue their doctoral studies in other countries. Although these improvements in nursing education in Iran are to be applauded, much more needs to be accomplished to ensure that highly competent nurse practitioners continue to be produced in this country. This article presents an historical overview of the development of nursing education in Iran, within its economic and sociopolitical contexts. Recommendations based upon lessons learned from historical and contemporary realities are presented in order to advance nursing education in this part of world. PMID- 17125891 TI - The development of national competency standards for the midwife in Australia. AB - OBJECTIVE: to develop and validate national competency standards for midwives in Australia. This study was part of a commissioned national research project to articulate the scope of practice of Australian midwives and to develop national competency standards to assist midwives to deliver safe and competent midwifery care. DESIGN: a multi-method, staged approach was used to collect data through a literature review, workshop consultations, interviews, surveys and written submissions in order to develop national competency standards for Australian midwives. Subsequently, direct observation of practice in a range of settings ensured validation of the competencies. SETTING: maternity-care settings in each state and territory in Australia. PARTICIPANTS: midwives, other health professionals and consumers of midwifery care. FINDINGS: The national competency standards for the midwife were developed through research and consultation before being validated in practice. KEY CONCLUSIONS: the national competency standards are currently being implemented into education, regulation and practice in Australia. These will be minimum competency standards required of all midwives who seek authority to practise as a midwife in Australia. It is expected that all midwives will demonstrate that they are able to meet the competency standards relevant to the position they hold. IMPLICATIONS FOR PRACTICE: the competency standards establish a national standard for midwives and reinforce responsibility and accountability in the provision of quality midwifery care through safe and effective practice. In addition, individual midwives may use the competency standards as the basis of their ongoing professional development plans. PMID- 17125892 TI - An Australian prospective cohort study of risk factors for severe perineal trauma during childbirth. AB - OBJECTIVE: to determine risk factors for the occurrence of severe perineal trauma (third and fourth degree tears) during childbirth. DESIGN: a prospective cohort study was conducted using the hospital's computerised obstetric information system. Additional data were gathered on women who sustained severe perineal trauma. Descriptive statistics and logistic regression were used to assess risk factors for severe perineal trauma. Midwives were asked to comment on possible reasons for severe perineal trauma. Written responses made by midwives were analysed using content analysis. Discussion groups with midwives were held to further explore their experiences. SETTING: Royal Prince Alfred Hospital, Sydney, Australia. PARTICIPANTS: all women having vaginal births (n=6595) in a 2-year period between 1 April 1998 and 31 March 2000, in both the birth centre and the labour ward. MEASUREMENTS AND FINDINGS: 2% of women (n=134) experienced severe perineal trauma. One hundred and twenty-two women had third-degree tears and 12 had fourth-degree tears. Primiparity, instrumental delivery, Asian ethnicity and heavier babies were associated with an elevated risk of severe perineal trauma. Midwives identified several factors they believed contributed to severe perineal trauma. These were lack of effective communication with the woman during the birth, different birth positions, delivery technique, ethnicity and obstetric influences. KEY CONCLUSIONS: findings support current knowledge that primiparity, instrumental birth, heavier babies and being of Asian ethnicity are associated with increased rates of severe trauma. Specific attention needs to be paid to the strong association found between being of Asian ethnicity and experiencing severe perineal trauma. IMPLICATIONS FOR PRACTICE: further identification and validation of the concerns expressed by midwives to reduce severe perineal trauma is warranted so that preventative strategies can be used and researched. PMID- 17125893 TI - Fifth metacarpal neck fractures: is follow-up required? AB - This study examines the premise that follow-up after fifth metacarpal neck fractures is unnecessary after initial assessment in a hand clinic. Forty consecutive patients with this fracture were managed in plaster and followed up. The next 38 patients were treated with neighbour strapping, given information sheets and no follow-up visits were arranged. In the first group, most return visits were for removal of plaster. Almost half of the appointments, thereafter, were not attended. In the second group, only two patients returned to the clinic, one with minor complaints and one with a repeat injury. Patients reported better satisfaction when managed with neighbour strapping (P=0.04) and without regular follow-up visits (P<0.01). The time to return to work was a mean of 5 weeks and 2.7 weeks in the first and second groups, respectively (P<0.01). There was no significant difference in the functional outcome at 12 weeks, as assessed by the DASH score. PMID- 17125894 TI - Synovial chondromatosis of the proximal interphalangeal joint. PMID- 17125895 TI - Workplace injury or "part of the job"?: towards a gendered understanding of injuries and complaints among young workers. AB - Epidemiological studies have found that teenage workers have higher occupational injury rates than adult workers, and that young males are a particularly high risk subgroup. However, there have been few studies to date that have explored qualitatively young workers' everyday understandings and experiences of occupational health risks. Based on focus groups conducted with Canadian urban and suburban teenagers aged 16-18 years, this paper explores young workers' understandings and experiences of occupational health risks, and their gendered nature. The respondents were employed in a diverse range of jobs. The findings suggest that young workers experience a number of minor injuries and physical complaints related to their work. These injuries were typically seen as "part of the job" because they happened frequently and were of low severity. Also, the experience of these injuries as "part of the job" was informed by the young workers' perceived lack of control to improve or alter the conditions of their work. Furthermore, young workers' complaints and concerns were systematically discounted and this happened in a gendered fashion. Whereas the females emphasized how their complaints were actively disregarded by their superiors, males (and some females in male-dominated work settings) described how they stifled their complaints in order to appear mature among their (older) co workers. Comparisons with qualitative studies of adult workers suggest that accepting some risks and injuries as "part of the job" is not peculiar to young workers. The implications of these findings for improving workplace safety for young workers are discussed. PMID- 17125896 TI - Why are you draining your brain? Factors underlying decisions of graduating Lebanese medical students to migrate. AB - In the context of a worldwide physician brain drain phenomenon, Lebanon has the highest emigration factor in the Middle East and North Africa. In this manuscript we aim to identify and develop a conceptual framework for the factors underlying the decisions of graduating Lebanese medical students to train abroad. We conducted two focus groups and seven semi-structured individual interviews with 23 students. In the deductive analysis (based on the push-pull theory), students reported push factors in Lebanon and pull factors abroad related to five dimensions. They focused predominantly on how training abroad provides them with a competitive advantage in an oversaturated Lebanese job market. An inductive analysis revealed the following emerging concepts: repel factors abroad and retain factors locally; societal expectations that students should train abroad; marketing of abroad training; and an established culture of migration. The marketing of abroad training and the culture of migration are prevalent in the academic institutions. PMID- 17125897 TI - China's transition: the effect of rapid urbanization on adult occupational physical activity. AB - China has recently undergone rapid social and economic change. Increases in urbanization have led to equally rapid shifts toward more sedentary occupations through the acquisition of new technology and transitions away from a mostly agricultural economy. Our purpose was to utilize a detailed measure of urbanicity comprising 10 dimensions of urban services and infrastructure to examine its effects on the occupational physical activity patterns of Chinese adults. Longitudinal data were from individuals aged 18-55 from the years 1991-1997 of the China Health and Nutrition Survey (N=4376 men and 4384 women). Logistic multilevel regression analyses indicated that men had 68% greater odds, and women had 51% greater odds, of light versus heavy occupational activity given the mean change in urbanization over the 6-year period. Further, simulations showed that light occupational activity increased linearly with increasing urbanization. After controlling for individual-level predictors, community-level urbanization explained 54% and 40% of the variance in occupational activity for men and women, respectively. This study provides empirical evidence of the reduction in intensity of occupational activity with modernization. It is likely that urbanization will continue unabated in China and this is liable to lead to further transitions in the labor market resulting in additional reductions in work-related activity. Because occupational activity remains the major source of energy expenditure for adults, unless alternative forms are widely adopted, the Chinese population is at risk of dramatic increases in the numbers of overweight and obese individuals. PMID- 17125898 TI - Activation of soluble guanylate cyclase by nitric oxide is increased in lymphocytes from both rats chronically exposed to 2,5-hexanedione and workers chronically exposed to n-hexane. AB - Although occupational exposure to n-hexane induces neurotoxic effects in the central and peripheral nervous systems, the mechanisms of its neurotoxicity remain unclear. n-Hexane is metabolized to 2,5-hexanedione (2,5-HD), which is the neurotoxic agent and the indicator chosen for the biological monitoring of exposed workers. It has been previously reported that chronic exposure to 2,5-HD impairs the glutamate-nitric oxide-cyclic GMP pathway at the level of activation of soluble guanylate cyclase (sGC) enzyme by nitric oxide (NO), both in cultured neurons and in the cerebellum of rats in vivo. The aim of this study was to assess whether the activation of sGC by NO is also altered in lymphocytes from rats treated with 2,5-HD and/or workers chronically exposed to n-hexane. Lymphocytes were isolated from male Wistar rats treated with 2,5-HD in drinking water, and from blood samples from shoe-factory workers environmentally and chronically exposed to n-hexane. Urine samples were also collected from workers at the end of the shift in order to measure the urinary levels of 2,5-HD. Activation of sGC by NO was significantly higher (p<0.05) in lymphocytes from rats treated with 2,5-HD than in control rats. In isolated lymphocytes from exposed workers the activation of sGC by NO also increases (p<0.05) in contrast to the controls. The results presented here indicate that the activation of lymphocytes could be an indicator of the toxicity produced by being exposed to n hexane, since the effects observed in workers chronically exposed to n-hexane are similar to those found in rats chronically treated with 2,5-HD in drinking water. PMID- 17125899 TI - The impact of montmorillonite clay addition on the in vitro wear resistance of a glass-ionomer restorative. AB - OBJECTIVES: The in vitro wear resistance of a glass-ionomer (GI) restorative ChemFil Superior (Dentsply DeTrey, Kanstanz, Germany) reinforced with either a pristine calcium montmorillonite (Ca-MMT) or an organically modified 12-amino dodecanoicacid treated montmorillonite (ADA-MMT) clay was evaluated to investigate the potential of MMT reinforced GI restoratives as posterior filling materials. METHODS: The wear resistance of a GI restorative reinforced with 0.5 2.5 wt.% Ca-MMT and ADA-MMT was assessed for groups of four disc-shaped specimens using the OHSU oral wear simulator operating at 20 N abrasion and 90 N attrition forces for 50,000 cycles at a frequency of 1 Hz in the presence of a poppy seed and PMMA micro-sphere slurry. The wear facet of each specimen was quantified by measuring the mean wear depth and mean volumetric wear in the abrasion (40-60% of wear facet) and attrition (80-90%) regions in conjunction with the mean total wear depth and mean total volumetric wear. RESULTS: The addition of Ca-MMT clay resulted in a significant increase in the mean total volumetric wear and mean total wear depth of the GI restorative as the concentration of clay was increased utilising the one-way ANOVA and Tukey's test comparisons at a 95% significance level. When ADA-MMT clay was added to the GI no significant differences in mean total volumetric wear and mean total wear depth were evident (P<0.05). CONCLUSIONS: The addition of Ca-MMT clay to the GI restorative resulted in a significant decrease in the in vitro wear resistance. ADA-MMT clay additions to the powder element of the GI restorative did not significantly enhance the in vitro wear resistance. PMID- 17125900 TI - Eicosapentaenoic acid modifies lipid composition in caveolae and induces translocation of endothelial nitric oxide synthase. AB - Endothelial nitric oxide synthase (eNOS) plays a crucial role in the regulation of a variety of cardiovascular functions. Many studies have shown that dietary n 3 polyunsaturated fatty acids (PUFAs) have beneficial effects on coronary atherosclerosis. However, the mechanisms of n-3 PUFAs regulation in eNOS activation remain unknown. In the present study we investigated the effects of eicosapentaenoic acid (EPA, 20:5 n-3) on subcellular distribution of eNOS and lipid composition of caveolae. We demonstrated for the first time that EPA treatment profoundly altered lipid composition and fatty acyl substitutions of phospholipids in caveolae. We found that caveolin-1 was solely located in caveolae fractions in control cells, and EPA treatment displaced caveolin-1 from caveolae. eNOS was detected in the caveolin-enriched fractions and noncaveolae fractions in control cells. EPA treatment induced the translocation of eNOS from caveolae fractions to soluble fractions. P-eNOS was also distributed in both fractions. After EPA treatment, the level of p-eNOS in each fraction was increased but the distribution of which was unaffected. Moreover, the results of immunofluorescence confirmed that EPA could redistribute caveolin-1 and eNOS in plasma membrane. eNOS activity in HUVEC cells was increased after EPA treatment, which was in a dose dependent manner. And incubation with 50 microM EPA had the maximum effect on eNOS activity. Our results suggested that eNOS translocation was paralleled by a stimulated capacity for NO production in the cells. We found that total Akt and p-Akt were primarily presented in heavy membranes in control cells, and the relative level of p-Akt increased but the distribution did not change after EPA treatment. The distribution of CaM was slightly changed after EPA treatment. Our results indicated that n-3 PUFAs profoundly altered caveolae microenvironment, thereby modifying location and function of proteins in caveolae. EPA-induced alterations of lipid and proteins in caveolae may be an important mechanism in the pathophysiologic process of atherosclerosis. PMID- 17125901 TI - Haemothorax: A potentially fatal complication of subclavian cannulation--A case report. AB - Central venous cannulation forms one of the mainstays of management of trauma and haemodynamically unstable patients. Frequently this procedure has to be done as an emergency in the emergency department. Ultrasound guidance has been recommended, especially for use by trainee members of the emergency department. We report a case of 44% full thickness burns with subclavian artery puncture due to a misplaced central venous catheter. This led to massive haemothorax, which eventually caused the patient to succumb. We re-emphasise the need for ultrasound guidance for insertion of central venous lines, especially in the emergency setting. PMID- 17125902 TI - The interplay of heuristics and parsing routines in sentence comprehension: evidence from ERPs and reaction times. AB - Semantic anomalies like "the fox that hunted the poacher" elicit P600 effects. Kolk et al. [Kolk, H.J., Chwilla, D.J., Van Herten, M., Oor, P.J.W., 2003. Structure and limited capacity in verbal working memory: a study with event related potentials. Brain and language, 85(1), 1-36] proposed that this P600 effect is triggered by a conflict between the outcome of a lexical strategy with that of the parsing routine. Specifically, when the lexical strategy indicates that the poacher hunted the fox, the full parse leads to the conclusion that the fox was the one who did the hunting. We tested this hypothesis by replicating the study cited above but manipulating the context by means of instruction. Participants were informed that semantic anomalies were created on purpose and that they should not be misled by these anomalies but instead focus on syntax or sentence structure. This instruction led to a strong reduction in P600 effect. This result supports the view that expectations play an important role in the generation of P600 effects to semantic anomalies, as proposed by Kolk et al. [Kolk, H.J., Chwilla, D.J., Van Herten, M., Oor, P.J.W., 2003. Structure and limited capacity in verbal working memory: a study with event related potentials. Brain and language, 85(1), 1-36]. PMID- 17125903 TI - Comparison of biological behavior between early-stage adenocarcinoma and squamous cell carcinoma of the uterine cervix. AB - OBJECTIVES: (1) To compare the anatomopathological variables and recurrence rates in patients with early-stage adenocarcinoma (AC) and squamous cell carcinoma (SCC) of the uterine cervix; (2) to identify the independent risk factors for recurrence. STUDY DESIGN: This historical cohort study assessed 238 patients with carcinoma of the uterine cervix (IB and IIA), who underwent radical hysterectomy with pelvic lymph node dissection between 1980 and 1999. Comparison of category variables between the two histological types was carried out using the Pearson's chi(2)-test or Fisher exact test. Disease-free survival rates for AC and SCC were calculated using the Kaplan-Meier method and the curves were compared using the log-rank test. The Cox proportional hazards model was used to identify the independent risk factors for recurrence. RESULTS: There were 35 cases of AC (14.7%) and 203 of SCC (85.3%). AC presented lower histological grade than did SCC (grade 1: 68.6% versus 9.4%; p<0.001), lower rate of lymphovascular space involvement (25.7% versus 53.7%; p=0.002), lower rate of invasion into the middle or deep thirds of the uterine cervix (40.0% versus 80.8%; p<0.001) and lower rate of lymph node metastasis (2.9% versus 16.3%; p=0.036). Although the recurrence rate was lower for AC than for SCC (11.4% versus 15.8%), this difference was not statistically significant (p=0.509). Multivariate analysis identified three independent risk factors for recurrence: presence of metastases in the pelvic lymph nodes, invasion of the deep third of the uterine cervix and absence of or slight inflammatory reaction in the cervix. When these variables were adjusted for the histological type and radiotherapy status, they remained in the model as independent risk factors. CONCLUSION: The AC group showed less aggressive histological behavior than did the SCC group, but no difference in the disease free survival rates was noted. PMID- 17125904 TI - In vitro investigation of the geometric contraction behavior of chemo-mechanical P-protein aggregates (forisomes). AB - We investigated the contracting behavior of forisomes from Vicia faba by carrying out precise measurements of their changing geometric parameters in vitro in the absence and in the presence of dissolved oxygen. Furthermore, we investigated the fine structure of forisomes by scanning electron microscopy. For the first time, single forisomes were titrated with Ca(2+), protons, and hydroxide ions recording the complete progression of their contractions. An apparent Ca(2+)-binding constant of (22+/-3) muM was calculated from two complete titration curves. The forisomes also contracted in the presence of Ba(2+) and Sr(2+) ions, but the amplitudes of contraction were smaller under the same measuring conditions. The time taken to change from the longitudinally expanded into the longitudinally contracted state was up to 2 s shorter in 10 mM Ca(2+) in comparison to 0.2mM Ca(2+). However, the contraction time was prolonged by decreasing the Ca(2+) concentration. In the absence of dissolved oxygen, the transition between the two final states of the forisomes was almost reversible and the amplitude of contraction remained almost constant during the first 25 contraction cycles. In the presence of dissolved oxygen the forisomes denaturated after a few cycles and lost their ability to contract, just after only a few cycles with 10 min in the contracted state. Denaturation of the forisomes occurred appreciably in the contracted state. We propose a cycle process to explain the thermodynamic basis of the Ca(2+)-induced contraction and its reversal by EDTA. Reducing the pH-value from 7.3 to 4.0 caused the forisomes to shorten by approximately 15%, while increasing the pH to 11.0 caused them to shorten by 28 to 30%. In both cases, the increases of the forisomes volume were greater than during the Ca(2+) induced contraction. The pH values of 4.7+/-0.3, and 10.2+/-0.2 marked the inflection points of the acid base titration of different forisomes. PMID- 17125905 TI - The costs and benefits of reforestation in Liping County, Guizhou Province, China. AB - Reducing greenhouse gases in the atmosphere is becoming a pressing issue for the global community. Afforestation and reforestation are promoted worldwide as an effective means of sequestering carbon. For its national interest and global concerns, China has made great efforts to protect its existing forests and develop programs of afforestation and reforestation. Based on two surveys recently conducted in Liping County, Guizhou province, this paper investigates the economic changes associated with the implementation of the "Grain For Green" policy. Based on the analytical framework of benefit cost analysis, this paper concludes that the implementation of the reforestation of sloping agricultural land policy would not be possible if there were no government subsidies for the peasants. The short term economic returns of land and labour from forestation are substantially lower than those generated from grain or cash crop production on the steep slope lands. The government subsidies provide great economic incentives for peasants to take part in the project. The subsidies in fact have elevated peasant income in rural Liping. The estimated potential economic returns of plantations over the long run indicate that the removal of the government financial subsidies would not create an economic crisis for the peasants if the current market conditions continue. PMID- 17125906 TI - Thermotherapy of prostate cancer using magnetic nanoparticles: feasibility, imaging, and three-dimensional temperature distribution. AB - OBJECTIVES: To investigate the feasibility of thermotherapy using biocompatible superparamagnetic nanoparticles in patients with locally recurrent prostate cancer and to evaluate an imaging-based approach for noninvasive calculations of the three-dimensional temperature distribution. METHODS: Ten patients with locally recurrent prostate cancer following primary therapy with curative intent were entered into a prospective phase 1 trial. The magnetic fluid was injected transperineally into the prostates according to a preplan. Patients received six thermal therapies of 60-min duration at weekly intervals using an alternating magnetic field applicator. A method of three-dimensional thermal analysis based on computed tomography (CT) of the prostates was developed and correlated with invasive and intraluminal temperature measurements. The sensitivity of nanoparticle detection by means of CT was investigated in phantoms. RESULTS: The median detection rate of iron oxide nanoparticles in tissue specimens using CT was 89.5% (range: 70-98%). Maximum temperatures up to 55 degrees C were achieved in the prostates. Median temperatures in 20%, 50%, and 90% of the prostates were 41.1 degrees C (range: 40.0-47.4 degrees C), 40.8 degrees C (range: 39.5-45.4 degrees C), and 40.1 degrees C (range: 38.8-43.4 degrees C), respectively. Median urethral and rectal temperatures were 40.5 degrees C (range: 38.4-43.6 degrees C) and 39.8 degrees C (range: 38.2-43.4 degrees C). The median thermal dose was 7.8 (range: 3.5-136.4) cumulative equivalent minutes at 43 degrees C in 90% of the prostates. CONCLUSION: The heating technique using magnetic nanoparticles was feasible. Hyperthermic to thermoablative temperatures were achieved in the prostates at 25% of the available magnetic field strength, indicating a significant potential for higher temperatures. A noninvasive thermometry method specific for this approach could be developed, which may be used for thermal dosimetry in future studies. PMID- 17125907 TI - Midterm follow-up of high-grade genital prolapse repair by the trans-obturator and infracoccygeal hammock procedure after hysterectomy. AB - OBJECTIVE: To prospectively assess the anatomic and functional outcome of high grade genital prolapse repair using a nonabsorbable hammock placement with anterior trans-obturator and posterior infracoccygeal extensions after hysterectomy. METHODS: Forty-four women with stage III or IV prolapse underwent surgery between January 2002 and June 2005. Patients had physical examination for prolapse assessment according to the International Continence Society pelvic organ prolapse staging system and were evaluated for subjective prolapse symptoms preoperatively and postoperatively. Follow-up was done at 6 wk and 6 mo and then once a year. RESULTS: Median follow-up was 29.3 mo (range: 9-47 mo). Thirty-six (81.8%) women had optimal anatomic results and seven (15.9%) had persistent asymptomatic stage I prolapse. One patient (2.7%) had symptomatic recurrence of a posterior vaginal wall stage III prolapse. The subjective cure rate was 97.7% (43 of 44). All subjective symptoms of prolapse decreased after surgery. The only intraoperative complication was an uneventful rectal injury. The rates of vaginal erosions and mesh infections were 13.6% and 4.5%, respectively. CONCLUSIONS: These results suggest that nonabsorbable hammock placement using anterior trans obturator and posterior infracoccygeal extensions could be a safe and effective treatment for high-grade genital prolapse. Further studies are warranted to determine long-term outcome and to compare this approach with previously accepted surgical procedures. PMID- 17125908 TI - What does the urologist expect from the pathologist (and what can the pathologists give) in reporting on adult kidney tumour specimens? AB - OBJECTIVE: To identify the parameters required by the urologist to determine the prognosis and the treatment of renal cancer in adults, and to establish the potential therapeutic targets of the new treatments that started to show clinical efficacy. METHODS: A literature search of the last 10 yr was done, paying specific attention to TNM 2002 (UICC staging) and Fuhrman's grading. Also, the main genetic characteristics of the different subtypes (according to the WHO 2004 classification) with potential therapeutic implications have been compiled. RESULTS: After the review of the literature, the opinion of the joint meeting including urologists and pathologists is that some aspects of the TNM 2002 classification must be refined. Criteria for nuclear grading should be different for the subtypes of renal cell carcinoma, and the WHO 2004 histological classification is clinically useful. CONCLUSIONS: In the workshop held in Palermo, common opinion was achieved on a number of points. The TNM 2002 classification is useful, but some adjustments should be made, particularly as referred to the tumour size cut-off, assessment of the invasion of the renal sinus fat tissue, and invasion of the ipsilateral adrenal gland. The Fuhrman's grading system is useful in clear cell renal cell carcinoma (RCC), and probably also in papillary RCC, but a redefinition for chromophobe RCC is needed. Finally, the determination of certain markers, such as VEGF and HIF, could constitute good target markers for the new therapies, but they remain under investigation. PMID- 17125909 TI - Transitional cell carcinoma of the ureter: prognostic factors influencing progression and survival. AB - OBJECTIVES: We retrospectively evaluated prognostic factors for progression-free and disease-specific survival in a large cohort of patients with transitional cell carcinoma (TCC) of the ureter. METHODS: A single-centre series of 145 consecutive patients treated with partial resection of the ureter or nephroureterectomy between 1975 and 2004 was evaluated. Median follow-up was 96 mo. Routine preoperative laboratory parameters as well as clinical and tumour specific data were assessed. Univariate and multivariate statistical analyses were performed. RESULTS: Five-year disease-specific survival ranged from 96.1% for stages pTa to 28.6% for pT4. Grade1 tumours were associated with 5-yr disease specific survival rates of 100% compared with 84% for G2, and 51.9% for G3 tumours, respectively. Univariate analyses identified pT stage and grade, tumour diameter, cM and pN categories, weight loss, the presence of synchronous tumour in the renal pelvis as well as elevated levels for humoral factors such as serum alkaline phosphatase (AP), white blood cell (WBC) count, platelet count, gamma glutamyl transferase, creatinin, and blood urea nitrogen as prognostic factors. In multivariate analyses, tumour grade and WBC counts were predictive for low progression-free survival rates, whilst simultaneous tumour in the renal pelvis, high AP levels, and WBC counts were correlated with worse disease-specific survival. CONCLUSIONS: Our retrospective analysis provides clinical factors to identify patients with TCC of the ureter at high risk for progression and death of disease. Interestingly, humoral factors such as elevated serum AP levels and high WBC counts were demonstrated to be of paramount prognostic significance besides tumour stage, grade and multifocality. PMID- 17125911 TI - KLF6 IVS1 -27G>A variant and the risk of prostate cancer in Finland. AB - OBJECTIVES: A recent report demonstrated that KLF6 IVS1 -27G>A substitution increases the transcription of alternatively spliced isoforms; this action was suggested to be associated with prostate cancer (pCA). To evaluate these findings among the Finnish population, a total of 3348 samples were analysed. METHODS: The variant was genotyped in 164 patients with familial pCA, 852 patients with unselected pCA, 459 patients with benign prostate hyperplasia (BPH), 923 male population controls, and 950 men from a Finnish prostate-specific antigen (PSA) screening trial with PSA levels less than 1.0ng/ml. Odds ratios (ORs) and corresponding 95% confidence intervals (95%CIs) were calculated by using logistic regression to estimate pCA risk. RESULTS: Association testing revealed no significant differences between familial prostate cancer patients and population controls (OR: 0.84; 95%CI, 0.56-1.28; p=0.42), unselected cases and controls (OR: 0.95; 95%CI, 0.76-1.19; p=0.63), or BPH cases and controls (OR: 1.12; 95%CI, 0.86 1.46; p=0.39). pCA and BPH cases were also compared with PSA-screened controls. None of these analyses revealed any significant associations. CONCLUSIONS: Our results do not support the suggested association of KLF6 IVS1 -27G>A germline polymorphism with pCA risk and also suggest that the variant is not a risk allele for BPH in the Finnish population. PMID- 17125912 TI - The natural history of noncastrate metastatic prostate cancer after radical prostatectomy. AB - OBJECTIVES: To characterise the natural history of metastatic prostate cancer after radical prostatectomy (RP) in patients followed expectantly for rising prostate-specific antigen (PSA) (noncastrate metastases). METHODS: Cox proportional hazards analyses were used to assess predictors of survival among 95 patients who developed clinically detectable noncastrate metastases after RP. The initial metastatic phenotype was characterised as minimal (nodal or axial skeletal involvement) or extensive (appendicular skeletal involvement or visceral metastases). Estimates of survival after diagnosis of metastases were generated with the Kaplan-Meier method. RESULTS: Median disease-specific survival from diagnosis of noncastrate metastases was 6.6 yr (95% confidence interval [CI], 5.2, 7.9). The initial site of metastatic disease was bone, lymph node, and viscera in 63%, 36%, and 6% of patients, respectively. Thirteen patients (14%) had extensive disease at their first metastatic manifestation. Longer PSA doubling time in the rising PSA state (hazard ratio [HR] 0.8 for each month increase in doubling time; 95%CI, 0.67-0.94) and the initial metastatic phenotype (HR 0.3 for minimal vs. extensive disease; 95%CI, 0.1-0.6) were associated with improved survival. The prostatectomy Gleason score, lymph node status at RP, PSA level at diagnosis of metastases, and interval from surgery to diagnosis of metastases did not correlate with outcome. CONCLUSION: Men who develop noncastrate metastases after RP may have a durable survival. Favourable prognostic indicators include longer PSA doubling time preceding diagnosis of metastases and initial involvement of axial skeleton or lymph nodes. PMID- 17125913 TI - Cadmium induces mitogenic signaling in breast cancer cell by an ERalpha-dependent mechanism. AB - Breast cancer (BC) is linked to estrogen exposure. Estradiol (E2) stimulates BC cells proliferation by binding the estrogen receptor (ER). Hormone-related cancers have been linked to estrogenic environmental contaminants. Cadmium (Cd) a toxic pollutant, acts as estrogens in BC cells. Purpose of our study was to evaluate whether Cd regulates MCF-7 cell proliferation by activating ERK1/2, Akt and PDGFRalpha kinases. Cd increased cell proliferation and the ER-antagonist ICI 182,780 blunted it. To characterize an ER-dependent mechanism, ERalpha/beta expression was evaluated. Cd decreased ERalpha expression, but not ERbeta. Cd also increased ERK1/2, Akt and PDGFRalpha phosphorylation while ICI blocked it. Since stimulation of phosphorylation was slower than expected, c-fos and c-jun proto-oncogenes, and PDGFA were analyzed. Cd rapidly increased c-jun, c-fos and PDGFA expression. Cells were also co-incubated with the Cd and specific kinases inhibitors, which blocked the Cd-stimulated proliferation. In conclusion, our results indicate that Cd increases BC cell proliferation in vitro by stimulating Akt, ERK1/2 and PDGFRalpha kinases activity likely by activating c-fos, c-jun and PDGFA by an ERalpha-dependent mechanism. PMID- 17125914 TI - Activation of mitogen-activated protein kinases by cisplatin and their role in cisplatin-resistance. AB - Mitogen-activated protein kinases (MAPKs) are critical components of a complex intracellular signalling network that ultimately regulates gene expression in response to a variety of extracellular stimuli. By transducing the signals from the cell surface to the nucleus and activating there gene expression, MAPKs control cell proliferation, differentiation and cell death. In mammalian cells there are three major pathways of MAPKs: stress-activated protein kinase/c-Jun-N terminal kinase (SAPK/JNK), p38 kinase and extracellular signal-regulated kinase (ERK). Generally, SAPK/JNK and p38 are key mediators of stress and inflammation responses evoked by a variety of physical, chemical and biological stress stimuli, while ERK 1/2 cascade is mostly induced by growth factors. The importance of MAPKs activation in cell response to cis diamminedichloroplatinum(II) (cisplatin; cDDP) and resistance development to this anti-cancer drug has been gradually appreciated in recent years. Today it is believed that MAPK activation is a major component deciding the cell fate in response to cisplatin. Their role in response to cisplatin is complex as these proteins, in most cases, are able to induce apoptosis, but also suppress it or have no role in this process. The final decision depends on the cell type, as well as proliferation and differentiation status of tumour cells. This review summarises current knowledge concerning the role of MAPK family members in cell response to cDDP, as well as their role in cisplatin-resistance. PMID- 17125915 TI - Bovine papillomavirus type-2 DNA and expression of E5 and E7 oncoproteins in vascular tumours of the urinary bladder in cattle. AB - Cattles suffering from chronic enzootic haematuria frequently develop urinary bladder tumours of both epithelial and mesenchymal origin mainly haemangioma and its malignant counterpart. The role of the bovine papillomavirus type-2 (BPV-2) and of its major transforming oncoprotein in naturally occurring urothelial carcinogenesis has been recently clarified. E5 interacts in vivo as in vitro with the beta receptor for the platelet-derived growth factor (PDGF). However, studies regarding tumours of mesenchymal origin such as those arising from blood vessels are lacking. We show that the BPV-2 is present in 100% of the vascular tumours of the urinary bladder examined. Twenty-six out of twenty-seven tumour samples (96%) expressed E5 while 20 out of 27 (74%) tumour samples expressed E7. The two viral oncoproteins were not expressed in normal endothelial cells. Additionally, they co-localize in neoplastic endothelial cells as demonstrated by confocal immunofluorescence. PDGFbeta receptor was also shown to be expressed and co localizes with E5 in neoplastic blood vessels. Our results demonstrate, for the first time, that the BPV-2 is present in high percentage in tumours of mesenchymal origin arising in its natural host. Furthermore, the expression of the two viral oncoproteins confirm that the virus may have a causative role in the neoplastic process. PMID- 17125916 TI - Cooperative cytotoxicity of methyl jasmonate with anti-cancer drugs and 2-deoxy-D glucose. AB - The anti-cancer agent methyl jasmonate (MJ) acts in vitro and in vivo against various cancer cell lines, as well as leukemic cells from chronic lymphocytic leukemia (CLL) patients. Given the importance of multi-agent combinations in cancer chemotherapy, the purpose of this study was to identify super-additive combinations of MJ and currently-available chemotherapeutic drugs. We identified such cooperative effects in six cell lines arising from different major types of malignancies, i.e., breast, lung, prostate and pancreas carcinomas as well as leukemia. The chemotherapeutic drugs tested were adriamycin, taxol, BCNU and cisplatin. For instance, MJ exhibited strong cooperative effects with BCNU in MIA PaCa-2 pancreatic carcinoma cells. Furthermore, MJ enhanced significantly (pV=0.028) the anti-leukemic effect of adriamycin in vivo, in a CLL mouse model. Finally, MJ cooperated with the glycolysis inhibitor 2-deoxy-D-glucose in inducing death of several types of carcinoma cells. We conclude that administration of MJ with common chemotherapeutic drugs and glycolysis inhibitors bears a promise for effective anti-cancer therapy. PMID- 17125917 TI - Oncogenic pathways of AML1-ETO in acute myeloid leukemia: multifaceted manipulation of marrow maturation. AB - The leukemic fusion protein AML1-ETO occurs frequently in human acute myeloid leukemia (AML) and has received much attention over the past decade. An initial model for its pathogenetic effects emphasized the conversion of a hematopoietic transcriptional activator, RUNX1 (or AML1), into a leukemogenic repressor which blocked myeloid differentiation at the level of target gene regulation. This view has been absorbed into a larger picture of AML1-ETO pathogenesis, encompassing dysregulation of hematopoietic stem cell homeostasis at several mechanistic levels. Recent reports have highlighted a multifaceted capacity of AML1-ETO directly to inhibit key hematopoietic transcription factors that function as tumor suppressors at several nodal points during hematopoietic differentiation. A new model is presented in which AML1-ETO coordinates expansion of the stem cell compartment with diminished lineage commitment and with genome instability. PMID- 17125918 TI - Simvastatin induces apoptosis in human breast cancer cells: p53 and estrogen receptor independent pathway requiring signalling through JNK. AB - The effect of simvastatin, a widely used statin for the treatment of hypercholesterolemia, was investigated in the estrogen receptor (ER)-positive MCF 7, and the ER-negative MDA-MB 231 human breast cancer cell lines. Simvastatin induced cell cycle arrest and apoptosis in both cells. These effects of simvastatin were not altered by 17-beta-estradiol treatment. MCF-7 cells express wild-type tumor suppressor protein p53, whereas MDA-MB 231 cells carry a p53 mutation. However, no alteration in the level or localisation of p53 was observed with simvastatin treatment in either cell line. On the other hand, simvastatin strongly stimulated phosphorylation of c-jun which was completely abolished by the c-jun NH2-terminal kinase (JNK) inhibitor SP600125, which also significantly reduced the antiproliferative and apoptotic effects of simvastatin in these cells. In conclusion, we describe here that simvastatin induces apoptosis via involvement of JNK in breast cancer cells independent of their ER or p53 expression status. These findings indicate a great potential for statins for the treatment of cancers resistant to currently used drugs, and target the JNK signalling pathway for a novel approach of breast cancer treatment. PMID- 17125919 TI - Degradation of azo dye Acid Orange 7 in water by Fe0/granular activated carbon system in the presence of ultrasound. AB - Degradation of azo dye Acid Orange 7 (AO7) was investigated using zero-valent iron/granular activated carbon (Fe(0)/GAC) in the absence and presence of ultrasound (US). The AO7 degradation efficiency by Fe(0)/GAC was dramatically enhanced by ultrasound, ultrasound alone had a little effect on the degradation. Effects of Fe(0)/GAC ratios and initial pH values on AO7 degradation were studied in the US-Fe(0)/GAC system. The degradation followed the pseudo-first-order kinetics model. The pseudo-first-order rate constant of AO7 degradation by Fe(0)/GAC was 8.74x10(-3)min(-1) while that by US-Fe(0)/GAC was 3.91x10(-2)min( 1). A significant synergetic effect was observed between US and Fe(0)/GAC. Ultrasound swept the reactive intermediates or products from Fe(0) surface, and thereby the surface of Fe(0) was cleaned and reactivated. Granular activated carbon was added as cathode to increase the unit of internal batteries and enhance the current efficiency of the internal electrolysis. The suitable Fe(0)/GAC ratio (v/v) for the degradation of AO7 in the presence of ultrasound was 1:1. The degradation was highly dependent on pH, and a decrease of the initial pH values from 12.0 to 4.0 led to the increase of degradation efficiencies. Sulfanilamide was proved as the main residual degradation product in the solution by the analysis of high-performance liquid chromatograms. PMID- 17125920 TI - Removal of methylene blue from aqueous solution by peat. AB - In this study, the removal of methylene blue (MB) from aqueous solution by peat was analyzed. The peat was collected from a peatland at Arroio do Silva Beach, in Santa Catarina state, in the south of Brazil. Adsorption was conducted using varied initial concentrations of the MB solutions and three different temperatures (35, 45 and 60 degrees C). An adsorption time of around 4.5h was sufficient to reach the equilibrium for all temperatures, in the concentration range studied. Percentage removal was greater for diluted solutions, but the absolute amount of MB adsorbed by the peat at equilibrium increased with an increase of the initial concentration, corroborating the efficacy of the material as an adsorbent. Temperature influenced slightly the reaction, which was endothermic. Results indicated a multi-layered process and the data were analyzed considering pseudo-first-order, pseudo-second-order and intraparticle diffusion approaches. The latter two mechanisms seem to be significant in the rate controlling step. PMID- 17125921 TI - A comparative study of quantum yield and electrical energy per order (E(Eo)) for advanced oxidative decolourisation of reactive azo dyes by UV light. AB - This paper evaluates the quantum yield and electrical energy per order (E(Eo)) efficiency of Reactive Orange 4 (RO4) and Reactive Yellow 14 (RY14) azo dyes by three advanced oxidation processes (AOPs). Both dyes were completely decolourised by all these processes. The relative decolourisation efficiencies of these processes were in the following order: Fe(2+)/H(2)O(2)/UV>UV/TiO(2)>UV/H(2)O(2). The low efficiency of UV/H(2)O(2) process is mainly due to low UV absorption by hydrogen peroxide at the 365nm. The figure of merit E(Eo) values showed that UV/H(2)O(2) process consumes more electrical energy than the other two processes. The electrical energy consumption is in the following order: UV/H(2)O(2)>UV/TiO(2)>Fe(2+)/H(2)O(2)/UV. At low initial dye concentration higher quantum yield was observed in UV/TiO(2) process, whereas in photo-Fenton process higher quantum yield was observed at high initial dye concentration. The structure of dye molecule also influences the quantum yield and E(Eo) value. PMID- 17125922 TI - Pinoline and N-acetylserotonin reduce glutamate-induced lipid peroxidation in retinal homogenates. AB - Glutamate is a neurotransmitter associated with oxidative retinal disorders. Pinoline (PIN) and N-acetylserotonin (NAS) are newly identified neural protectors. We investigated the glutamate-induced lipid peroxidation (LPO) and the protective effects of PIN and NAS in the retina. Porcine retinal homogenates were treated with different concentrations of glutamate. The malondialdehyde (MDA) level per unit weight of protein was quantified spectro-photometrically as an index of LPO. The glutamate concentration that induced a significant increase in retinal MDA was determined. The glutamate-treated retinal homogenate was then co-incubated with 5 different concentrations (0, 35.7, 71.5, 143 and 286 microM) of PIN, NAS or their combinations (concentration corresponding to 25, 50 and 75% of protection). Glutamate induced a significant dose-dependent increase in retinal MDA (p<0.0001). Co-incubation with PIN or NAS significantly suppressed the glutamate-induced MDA (p<0.01) in a dose-dependent manner (p<0.0001). The concentrations to inhibit 50% of LPO were 132.8 and 98.6 microM for PIN and NAS, respectively. In summary, elevated glutamate induced retinal LPO. Both PIN and NAS suppressed the glutamate-induced LPO and a synergic protection was evident after incubation in PIN/NAS mixtures. PMID- 17125923 TI - GDNF reduces oxidative stress in a 6-hydroxydopamine model of Parkinson's disease. AB - Many current theories of Parkinson's disease (PD) suggest that oxidative stress is involved in the neurodegenerative process. Potential neuroprotective agents could protect neurons through inherent antioxidant properties or through the upregulation of the brain's antioxidant defenses. Glial cell line-derived neurotrophic factor (GDNF) has been shown to protect and restore dopamine neurons in experimental models of PD and to improve motor function in human patients. This study was designed to investigate GDNF's effect on oxidative stress in a model of PD. GDNF or vehicle was injected into the right striatum of male Fischer 344 rats. Three days later 6-OHDA or saline was injected into the same striatum. The striatum and substantia nigra from both sides of the brain were removed 24h after 6-OHDA or saline injection and analyzed for the oxidative stress markers protein carbonyls and 4-hydroxynonenal. Both markers were significantly reduced in GDNF+6-OHDA treated animals compared to vehicle+6-OHDA treated animals. In addition, in animals allowed to recover for 3.5-4 weeks after the 6-OHDA administration, the GDNF led to significant protection against loss of striatal and nigral tissue levels of dopamine. These results suggest that the protective effects of GDNF against 6-OHDA involve a reduction in oxidative stress. PMID- 17125924 TI - Pre-activating the mesial temporal lobe facilitates learning. AB - While numerous neuroimaging studies have been conducted to examine brain activities associated with different cognitive tasks, few have focused on the effects of temporary increase in blood oxygen level on subsequent cognitive performance. The present study aimed to study this temporary hemodynamic change on subsequent learning. Using the novel picture encoding task (NPET) to activate the hippocampus, the present functional magnetic resonance imaging study showed that the level of blood oxygen level-dependent signals in the mesial temporal lobe associated with NPET positively predicted memory performance after engaging in the NPET for normal controls and patients with temporal lobe epilepsy. In addition, patients demonstrated significantly greater improvement on memory performance than normal controls. Though the improvement was transient, these results provided a neuro-physiological evidence to support our previous hypothesis that the functioning of a specific brain region can be facilitated by performing a task mediated by the same region. PMID- 17125925 TI - Elaborate force coordination of precision grip could be generalized to bimanual grasping techniques. AB - Exceptional coordination of grip (G; the normal force that prevents slippage of the grasped object) and load force (L; the tangential force originating from the object's weight and inertia) has been interpreted as a part of evidence that both the anatomy and neural control of human hands have been predominantly designed for manipulation tasks. In the present study, we tested the hypothesis that the precision grasp (uses only the tips of fingers and the thumb of one hand) provides better indices of G and L coordination in static manipulation tasks than two bimanual grasps (palm-palm and fingers-thumb; both using opposing segments of two hands). However, in addition to a subtle difference in relative timing of G and L between the precision and two bimanual grasps, we only found that the fingers-thumb grasp is characterized with higher G/L ratio and somewhat higher modulation of G than not only the precision, but also the bimanual palm-palm grasp. However, all remaining data including the correlation coefficients between G and L demonstrated no difference among three evaluated grasping techniques. Therefore, we concluded that the elaborate G and L coordination associated with uni-manual grasps could be partly generalized to a variety of manipulation tasks including those based on bimanual grasping techniques. Taking into account the importance of manipulation tasks in both everyday life and clinical evaluation, future studies should extend the present research to both other grasping techniques and dynamic manipulation conditions. PMID- 17125926 TI - Sex steroids modulate the signals from volatile female odors in the accessory olfactory bulb of male mice. AB - We previously reported that male mice detect volatile female odors via the accessory olfactory system, and that these odors activate granule cells in the accessory olfactory bulb (AOB) with a characteristic pattern. We also reported that sex steroids modulate the attraction of male mice to volatile female odors. The present study investigated hormonal modulation of signals from volatile female odors in the AOB with c-Fos immunostaining. After intact male mice were exposed to volatile female odors, there were more c-Fos positive cells in the caudal granule cell layer (GCL) than in the rostral GCL of the AOB. This effect was observed 3 days but not 7 days after castration, suggesting that hormonal deficiency causes the reorganization of the AOB after 3 days. There was no difference in the number of c-Fos positive cells between the rostral and caudal GCL of castrated male mice treated with 17 beta-estradiol (E). In contrast, there were more c-Fos positive cells in the caudal GCL than in the rostral GCL of castrated male mice treated with dihydrotestosterone (DHT). In both DHT- and E treated castrated male mice, there was no difference in the number of c-Fos positive cells between the rostral GCL and caudal GCL. This finding suggests that E disrupts the effect of DHT, and that androgen is required for maintaining the intact neuronal network of the AOB. The present study suggests that sex steroids modulate the signals from volatile female odors in the AOB of male mice. PMID- 17125927 TI - Changes of cholinesterase activities in the plasma and some tissues following administration of L-carnitine and galanthamine to rats. AB - Changes of acetylcholinesterase (AChE) activities in the hypophysis and brain (frontal cortex, hippocampus, medial septum and basal ganglia), and butyrylcholinesterase in plasma and liver following galanthamine (GAL) administration were studied in rats pretreated with L-carnitine (CAR). Following only GAL administration (10 mg/kg, i.m.), both cholinesterases (without clinical symptoms of GAL overdosage) were significantly inhibited. Pretreatment with CAR (3 consecutive days, 250 mg/kg, p.o.) followed by GAL administration showed higher AChE inhibition in comparison with single GAL administration. However, a statistically significant difference was observed for AChE in the hippocampus only. The activity of peripheral cholinesterases was not influenced by CAR pretreatment. Thus, pretreatment with CAR enhanced AChE inhibition in some brain parts of the rat following GAL administration. PMID- 17125928 TI - Screening for Echinococcus granulosus in dogs: Comparison between arecoline purgation, coproELISA and coproPCR with necropsy in pre-patent infections. AB - Echinococcus granulosus is an important zoonotic infection of dogs. The purpose of the present study assessed the performance of two laboratory diagnostic methods with arecoline purgation and necropsy in infected dogs. In total 65 dogs were successfully experimentally infected with protoscoleces of E. granulosus from ovine infection. At 14-34 days post-infection groups of dogs were purged with arecoline hydrobromide and then necropsied. Faecal samples were tested at weekly intervals by coproantigen ELISA and coproPCR. The necropsy infection rate with E. granulosus was 89.2 per cent. Only 43 per cent of dogs were successfully purged after one arecoline dose; this percentage increased to 76.9% for two doses of arecoline purgation. E. granulosus coproantigen was detected by coproELISA in 82.8% of faeces. The positive and negative predictive values for coproantigen ELISA were 96 and 44.4% respectively. E. granulosus DNA was detected in pre patent faecal samples by coproPCR in 25.9% of dogs. These results indicate that coproELISA is more sensitive than arecoline purgation for the detection of pre patent E. granulosus infection in dogs. CoproPCR detected E. granulosus DNA in dog faeces by 21 days post-infection before egg production. PMID- 17125929 TI - Recombinant galectins of male and female Haemonchus contortus do not hemagglutinate erythrocytes of their natural host. AB - Recombinant galectins of female and male adult worms of Haemonchus contortus were expressed in Escherichia coli and their hemagglutinating activities to human and different animal erythrocytes were analyzed. The results showed that female and male galectins could be highly expressed in E. coli using a temperature-sensitive plasmid, with the recombinant protein being mainly appeared in inclusion bodies. Hemagglutinating activity assays showed that both of the galectins hemagglutinated human A, B, O type, dog, rabbit, chicken and mouse erythrocytes at the high concentration of 40 microg/well, but did not hemagglutinate erythrocytes of the natural host of H. contortus, the goat. Sugar inhibition assays confirmed that, out of eight sugars tested, only lactose was effective to inhibit agglutination of human type B erythrocytes by the recombinant galectins. PMID- 17125930 TI - Effects of early enteral arginine supplementation on resuscitation of severe burn patients. AB - OBJECTIVES: To investigate the effects of dietary supplementation of l-arginine (l-Arg) on shock in severely burned patients. METHODS: This was a prospective, randomized, single blind, controlled study. Forty-seven severely burned patients due to various causes with a total burn surface area (TBSA) more than 50% each admitted in early postburn phase (within 10h postburn) were included in this study. All patients were treated by the traditional resuscitation program of our institute. After the nasogastric feeding tube was placed, they were randomly divided into three groups-(1) group A400 (n = 16): giving gastrointestinal feeding with 500 ml 5% GNS, containing l-Arg (400 mg/ kgday) at equal pace with fluid resuscitation; (2) group A200 (n = 16): giving gastrointestinal feeding with 500 ml 5% GNS containing l-Arg (200 mg/ kgday); (3) group C (n = 15): giving gastrointestinal feeding with 500 ml 5% GNS without any supplementation. The feeding started within 12h after burn and lasted for 72 h, the feeding rate was controlled by an enteral feeding pump. The following parameters were observed on days (PBD) 1-4: serum nitric oxide content (NO), mean arterial blood pressure (MAP), oxygenation index (PO2/FiO2), and arterial blood content of lactic acid (LA). Gastric mucosal blood flow was measured by laser Doppler flow-metry on PBD1 and PBD2. RESULTS: (1) Enteral feeding of l-Arg did not change MAP of severely burned patients, with no difference in MAP between the l-Arg supplemented and control groups. (2) There were significant changes of the l-Arg supplemented groups (A400 and A200), with an increased gastric mucosa blood flow, oxygenation index, and a decreased LA content in arterial blood, compared with the control group. (3) The serous NO content was significantly decreased in the A400 group on PBD2-4 (P < 0.01), and in the A200 group on PBD4 (P < 0.05) compared with the control group. CONCLUSIONS: Enteral feeding with l-arginine supplementation on early stage of burn decreases NO production to a relatively normal level and exerts beneficial effects on the resuscitation of burned shock. PMID- 17125931 TI - Relationship between self-reported task persistence and history of quitting smoking, plans for quitting smoking, and current smoking status in adolescents. AB - The task persistence construct has previously been measured primarily behaviorally (e.g., with a mirror-tracing task, or breath holding), and only in adults. It has been shown to differentiate between adult smokers and non-smokers and to predict smoking cessation in adult smokers trying to quit. This theory based analysis is the first to examine task persistence in adolescent smokers and to examine a two-item, internally consistent, self-report measure of task persistence. Results indicate that task persistence is greater among adolescent non-smokers as compared to adolescent current smokers, and those planning to quit smoking as compared to those with no plans to quit. Contrary to hypotheses, task persistence was not found to be related to prior successful attempts to quit smoking. Our results suggest that a brief, self-report measure of task persistence may be a methodologically sound, practical clinical tool for this population. PMID- 17125932 TI - Explicit and implicit alcohol-related cognitions and the prediction of future drinking in adolescents. AB - Both implicit and explicit alcohol-related cognitions might play a role in the early development of addictive behaviors. In this study, the association between both current and prospective alcohol use and implicit and explicit alcohol related cognitions were measured in two different adolescent age groups (N=100; 51 twelve year olds, 49 fifteen year olds). Alcohol-related cognitions were measured on two dimensions (valence and arousal). A new measure, the unipolar Single Target Implicit Association Test (ST-IAT), was used as the implicit measure. A unipolar expectancy questionnaire was used as the explicit measure. Current alcohol use and alcohol use after one year were measured with an alcohol use questionnaire. Abstainers and drinkers differed in both their explicit and implicit alcohol-related cognitions moderated by age and gender. Additionally, a hierarchical regression analysis showed that implicit associations with alcohol added significantly to the prediction of prospective binge drinking, when controlling for grade, gender and explicit alcohol expectancies These results indicate the importance of taking implicit alcohol-related cognitions into account when intervention methods are developed. PMID- 17125933 TI - Suspension culture combined with anticancer regimens for screening breast cancer stem cells. AB - Breast cancer stem cells (BrCSCs)have been first identified in solid tumors. To be more exact, they are defined as tumorigenic cancer cells which exhibit the capacity to form tumors distinct from the majority non-tumorigenic cancer cells. However it is only a minor subset that cancer stem cells are highly enriched in. It is still unknown how to find the definitive research subject - "a single breast cancer stem cell" - that can cause a new tumor by itself. Since recent studies suggest that BrCSCs have a higher proportion in suspension culture and have a greater resistance to chemotherapy regimens, we propose a novel strategy to isolate and identify BrCSCs:suspension culture combined with anticancer regimens. This double sorting method by means of the unique properties of BrCSCs may be helpful to screen genuine cancer stem cells(CSCs). PMID- 17125934 TI - A built-in co-carcinogenic effect due to viruses involved in latent or persistent infections. AB - A new hypothesis for some cancers, which combines the chromosomal instability theories with a co-carcinogenic effect of viruses causing latent or persistent infection, is presented. The hypothesis incorporates the multi-step model of cancer and that pre-cancerous cells reach a state of chromosomal instability. Because of chromosomal instability, the genome of these cell lines will lead to changes from generation to generation and will face a remarkable selection pressure both from lost traits, apoptosis, and from the immune system. Viruses causing latent or persistent infections have evolved many different genes capable to evade the immune system. If these viruses are harboured in the genome of pre cancerous cells they could provide them with "superpowers" and with genes that may assist the cells to elude the immune system. The theory explains why cancer predominantly is a disease of old age. Upon aging, the immune system becomes reduced including the ability to control and suppress the viruses that cause latent or persistent infections. The risk of cancer could thereby increase as the immune functions decrease. The theory provides new insights to the genesis of cancers. PMID- 17125935 TI - Could diabetic retinopathy be an autoimmune disease? AB - Diabetic retinopathy is a common and progressive complication of diabetes mellitus. It is characterized by the loss of pericytes, hypertrophy of basement membrane, microaneurysms formation, increased vascular permeability, capillary occlusions, neovascularisation and fibrovascular proliferation. The pathogenesis of diabetic retinopathy is still insufficiently understood, although some reports have implicated the role of the immune system. We hypothesize that, according to some current data diabetic retinopathy could also be considered as an autoimmune disease. The finding of antipericyte and antiendothelial cell autoantibodies in the circulation of diabetic patients strongly suggests that some autoimmune activity has been involved in the early pathophysiology of diabetic retinopathy. There is even more evidence that implicates the presence of autoimmune mechanisms in the proliferative stage of this disease: elevated levels of tumor necrosis factor-alpha, interleukin-8 and soluble interleukin-2 receptor in the serum of diabetic patients, increased vitreous concentration of the interleukin-6 and interleukin-8 in patients with proliferative retinopathy. Furthermore, preretinal membranes in diabetic patients contain deposits of immunoglobulins, activated complement components, monocytes, T and B lymphocytes, fibroblastes and lymphokynes. In diabetic patients human leukocyte antigen DR and DQ expression on the retinal vascular endothelial cells as well as on pigment and nonpigment epithelial cells was found. These antigens are normally restricted to immunocompetent cells and play an important regulatory role in the immune response. Their aberrant expression has been found on nonlymphoid cells in various autoimmune diseases whilst abnormal expression of DR and DQ antigens at sites where they do not normally exist would result in autoimmunity by converting the target cell into a functional antigen-presenting cell. In conclusion, although the pathogenesis of diabetic retinopathy is not completely understood it is known that the immune system is certainly involved in its development. However, there is increasing evidence of the presence of some autoimmune processes in the early stages of diabetic retinopathy and particularly in its proliferative phases. Consequently, diabetic retinopathy could also be considered as an autoimmune disease. PMID- 17125936 TI - Chronic benzylpiperazine (BZP) exposure produces behavioral sensitization and cross-sensitization to methamphetamine (MA). AB - BACKGROUND: Like other psychostimulant drugs, acute exposure to benzylpiperazine (BZP) increases dopaminergic neurotransmission, producing hyperactivity and stereotypy. The consequences of repeated BZP exposure have not however been investigated. The effects of acute and repeated BZP and methamphetamine (MA) exposure on locomotor activity and stereotypy were measured in order to determine whether there was sensitization and cross-sensitization between these two psychostimulant drugs. METHODS: The effects of acute treatment with MA (0.0, 0.5, 1.0 and 2.0 mg/kg, intraperitoneal (IP)) or BZP (0.0, 5.0, 10.0, 20.0 and 40.0 mg/kg, IP) on locomotor activity and stereotypy were determined. Effects of repeated exposure were determined in other groups that received five daily injections of 2.0 mg/kg MA, 20.0 mg/kg BZP or vehicle. Following a 2-day withdrawal period, rats from each treatment group received either a low dose MA (0.5 mg/kg) or BZP (10.0 mg/kg). RESULTS: MA and BZP produced dose-dependent hyperactivity and stereotypy. Repeated MA and BZP resulted in a potentiated locomotor but not stereotypy response. Following the withdrawal period, MA pretreated rats exhibited a sensitized locomotor and stereotypy response to the low dose MA and a conditioned response to saline. BZP pretreated rats also demonstrated a sensitized locomotor response to the low dose of BZP and MA. CONCLUSIONS: The present findings indicate that repeated exposure to BZP results in sensitization and cross-sensitization to MA. PMID- 17125937 TI - Expression of PACT is regulated by Sp1 transcription factor. AB - PACT is a stress-modulated, cellular activator of interferon (IFN)-induced double stranded (ds) RNA-activated protein kinase (PKR) and is an important regulator of PKR-dependent signaling pathways. The research presented here is aimed at understanding the regulation of PACT expression in mammalian cells. PACT is expressed ubiquitously in different cell types at varying abundance. We have characterized the sequence elements in PACT promoter region that are required for its expression. Using deletion analysis of the promoter we have identified the minimal basal promoter of PACT to be within 101 nucleotides upstream of its transcription start site. Further mutational analyses within this region, followed by electrophoretic mobility shift analyses (EMSAs) and chromatin immunoprecipitation (ChiP) analysis have shown that Specificity protein 1 (Sp1) is the major transcription factor responsible for PACT promoter activity. PMID- 17125938 TI - Catalytic activity of hammerhead ribozymes in a clay mineral environment: implications for the RNA world. AB - The hypothesized RNA-based world would have required the presence of a protected environment in which RNA, or an RNA-like molecule, could originate and express its biological activity. Recent studies have indicated that RNA molecules adsorbed/bound on clay minerals are able to persist in the presence of degrading agents, to interact with surrounding molecules, and to transmit the information contained in their nucleotide sequences. In this study, we assessed the ability of RNA molecules with catalytic activity to perform a specific reaction in a mineral environment. For this purpose, we investigated the self-cleavage reaction of the hammerhead ribozyme of the Avocado Sun Blotch Viroid (ASBVd), both in the monomeric and in dimeric forms. The monomeric transcript was tightly bound on the clay mineral montmorillonite to form a stable complex, while the behaviour of the dimeric transcript was studied in the presence of the clay particles in the reaction mixture. The results indicated that the hammerhead ribozyme was still active when the monomeric transcript was adsorbed on the clay surface, even though its efficiency was reduced to about 20% of that in solution. Moreover, the self-cleavage of clay-adsorbed molecule was significantly enhanced ( approximately four times) by the presence of the 5' reaction product. The self cleavage reaction of the dimeric transcript in the presence of montmorillonite indicated that the mineral particles protected the RNA molecules against aspecific degradation and increased the rate of cleavage kinetics by about one order of magnitude. These findings corroborate the hypothesis that clay-rich environments would have been a good habitat in which RNA or RNA-like molecules could originate, accumulate and undergo Darwinian evolutionary processes, leading to the first living cells on Earth. PMID- 17125939 TI - Application of RNA interference for inhibiting the replication of feline immunodeficiency virus in chronically infected cell lines. AB - RNA interference (RNAi) is a process in which double-stranded RNA induces the post-transcriptional sequence-specific degradation of homologous messenger RNA. The present study was carried out to apply the RNAi technology to inhibit the replication of feline immunodeficiency virus (FIV). Four small interfering RNAs (siRNAs) homologous to the FIV gag gene were synthesized and transfected into a feline fibroblastic cell line chronically infected with FIV (CRFK/FIV). These synthetic siRNAs efficiently inhibited the replication of FIV. Next, we examined the effect of retroviral vector-mediated transfer of FIV-specific short hairpin RNA (shRNA) on the replication of FIV in a feline T-cell line chronically infected with FIV (FL4). The retroviral vector-mediated transfer of FIV-specific shRNA was shown to markedly inhibit the replication of FIV in the FL4 cells. These results provide useful information for the development of RNAi-based gene therapy strategy to control FIV infection. PMID- 17125940 TI - Prevalence of ibeA gene in avian pathogenic Escherichia coli (APEC). PMID- 17125941 TI - Diallyl sulfide induces the expression of nucleotide excision repair enzymes in the breast of female ACI rats. AB - Diethylstilbestrol (DES) causes DNA adducts resulting in breast cancer, whereas diallyl sulfide (DAS) inhibits cancer formation. We hypothesize that DAS induces the expression of nucleotide excision repair genes. To test this hypothesis, female ACI rats were treated for 4 days with corn oil, DES, DAS, and DAS/DES (50mg/kg). The expression of P53, Gadd45a, PCNA, and DNA polymerase delta was analyzed by real-time PCR. DES decreased the expression of P53, Gadd45a and PCNA. DAS and DAS/DES increased the expression of all four genes. These results suggest that DAS enhances the ability of breast tissue to repair DNA damage thus preventing cancer. PMID- 17125942 TI - Effect of ambient temperature on in vitro fertilization of Bubaline oocyte. AB - The present study was used to examine the effect of ambient temperature on the day of slaughter of buffaloes on oocyte cleavage and subsequent embryo development following in vitro fertilization (IVF)/chemical activation (parthenogenesis). A total of 601 oocytes were collected from buffaloes, which were sacrificed when the ambient temperature was >40 or < or =40 degrees C and the collected oocytes were matured in vitro. During each experiment about half of the matured oocytes were used for IVF whereas the remaining oocytes were subjected to one of the three chemical activation protocols viz. (i) 7% ethanol (ET) and 6-di methyl amino purine (6-DMAP), (ii) ET and cycloheximide (CHX) and (iii) ET followed by a combined treatment of 6-DMAP and CHX. Cleaved oocytes were cultured in mSOF supplemented with BSA, essential amino acids, non-essential amino acids, ITS (insulin transferrin and selenium) and L-glutamine. Low cleavage and subsequent embryo development was observed in those oocytes which were collected from buffaloes slaughtered at ambient temperature >40 degrees C than at < or =40 degrees C. There was no significant difference in cleavage rate following different chemical activations in oocytes collected from buffaloes slaughtered on the day when the maximum ambient temperature was >40 degrees C or < or =40 degrees C. These results suggest that high ambient temperature influences competence of oocytes to cleave and develop to blastocyst stage following natural activation with sperm and/or process of fertilization and subsequent embryo development. PMID- 17125943 TI - Ethanolic extracts of black cohosh (Actaea racemosa) inhibit growth and oestradiol synthesis from oestrone sulphate in breast cancer cells. AB - Extracts of black cohosh (Actaea racemosa) and soy are used as 'natural' alternatives to conventional hormone replacement therapy (HRT) and there is some evidence that soy may protect against breast cancer by inhibiting the production of active oestrogens. This study compares the action of ethanolic extracts of black cohosh (BCE) and genistein on growth and enzyme activity in MCF-7 and MDA MB-123 breast cancer cells. BCE inhibited growth at the two highest doses tested, i.e. 50 and 100 microg/ml, whilst genistein stimulated growth in the oestrogen receptor positive (ER(+)) MCF-7 cells, but at high doses it inhibited growth in both cell lines. BCE did not affect the conversion of androstenedione to oestradiol and only the highest doses (50 and 100 microg/ml) significantly inhibited the conversion of oestrone to oestradiol in MDA cells. In contrast, BCE induced a dose-dependent inhibition of the conversion of oestrone sulphate to oestradiol in both cell lines, whilst in human granulosa lutein (GL) cells enzyme activity was only inhibited at the highest dose of BCE. Genistein had no significant effect on enzyme activity in breast cancer cells and like BCE only the highest doses (10 and 50 microM) inhibited enzyme activity in human GL cells. In vivo genistein may have growth stimulatory effects on breast tissue but BCE not only inhibits growth but inhibits the conversion of oestrone sulphate to active oestradiol, considered by some, to be the preferred pathway of oestradiol synthesis in breast tissue. PMID- 17125944 TI - Insect management products from Malian traditional medicine--establishing systematic criteria for their identification. AB - In material-resource poor countries like Mali, traditional practices incorporate the use of plants for medicinal purposes. Ethnobotanical research has documented traditional uses of plants, while concomitant studies by natural product chemists, ethnobotanists, and microbiologists have verified the efficacy of using traditional medicinal plants that have proven antimicrobial activity. These plants may also be used to protect agricultural crops pre-harvest and post harvest from insect herbivory. In Mali, subsistence farmers, regional scientists, and extension specialists rely on local plants for many medicinal needs and are amenable to using traditional plant materials for insect pest management. The goal of this research was to develop Integrated Pest Management (IPM) strategies using Malian traditional medicine as a discovery lead. The discovery premise was based on identifying plants through a matrix approach utilizing agricultural scientists, traditional practitioners, and subsistence farmers. We hypothesized that plants used in traditional medicine with antimicrobial activity lead to potential insect pest management agents. To test our hypothesis, we developed a four-step process for selecting Malian plant species. Seven criteria were selected to create a systematic matrix to identify the most promising plant materials for practical, affordable, ecologically-sound insect management by Malian farmers. In the first step of the process, we developed a list of 294 medicinal Malian plant species which were evaluated using the matrix. Sixty-seven plant species met our main criteria. After the environmental soundness of these species was evaluated using four minor criteria, 50 species emerged from this pre chemical, pre-bioassay process for further consideration in IPM programs in Mali. PMID- 17125945 TI - Antiulcer and in vitro antioxidant activities of Jasminum grandiflorum L. AB - The study was aimed at evaluating the antiulcer and antioxidant activities of 70% ethanolic axtract of leaves of Jasminum grandiflorum L. (JGLE). The leaves of Jasminum grandiflorum L. (Family: Oleaceae) is used in folk medicine for treating ulcerative stomatitis, skin diseases, ulcers, wounds, corns - a hard or soft hyperkeratosis of the sole of the human foot secondary to friction and pressure (Stedman's Medical Dictionary, 28th ed. Lippincott Williams & Wilkins, Philadelphia. p. 443), etc., Antiulcerogenic activity of JGLE (100 and 200 mg/kg, b.w., orally) was evaluated employing aspirin + pylorus ligation (APL) and alcohol (AL) induced acute gastric ulcer models and ulcer-healing activity using acetic acid-induced (AC) chronic ulcer model in rats. Both the antisecretory and cytoprotection hypothesis were evaluated. The antioxidant activity of JGLE has been assayed by using in vitro methods like 2,2-diphenyl-1-picrylhydrazylhydrate (DPPH) assay, reductive ability, superoxide anion scavenging activity, nitric oxide scavenging activity and total phenolic content, in order to explain the role of antioxidant principles in the antiulcerogenic activity of the extract. There was a significant (P<0.01) dose-dependent decrease in the ulcerative lesion index produced by all the three models in rats as compared to the standard drug famotidine (20 mg/kg, b.w. orally). The reduction in gastric fluid volume, total acidity and an increase in the pH of the gastric fluid in APL rats proved the antisecretory activity of JGLE. Additionally, JGLE completely healed the ulcer within 20 days of treatment in AC model as evidenced by histopathological studies. Like antiulcer activity, the free radical scavenging activities of JGLE depends on concentration and increased with increasing amount of the extract. These results suggest that leaves of Jasminum grandiflorum possess potential antiulcer activity, which may be attributed to its antioxidant mechanism of action. PMID- 17125946 TI - Antioxidant, anti-inflammatory, anti-nociceptive activities and composition of Lythrum salicaria L. extracts. AB - Lythrum salicaria (purple loosestrife) known as "Tibbi hevhulma" in Turkish is used for its several beneficial health effects against as diarrhea, chronic intestinal catarrh, hemorrhoid and eczema in the form of a decoction or a fluid extract and to treat varicose veins, bleeding of the gums, hemorrhoid and eczema, externally. Dried herbal parts of Lythrum salicaria L. (Lythraceae) were sequentially extracted with different solvents such as petroleum ether, ethyl acetate, methanol and 50% aqueous methanol. Water extract of Lythrum salicaria was also prepared under reflux. Antioxidant, anti-inflammatory and anti nociceptive activities of all the extracts were investigated using in vitro and in vivo methods, respectively. Free radical scavenging activity (1,1-diphenyl-2 picrylhydrazyl, DPPH* assay), iron(III) reductive activity, capacity of the inhibition of linoleic acid peroxidation and MDA formation, anti-nociceptive activity (p-benzoquinone-induced abdominal constriction test) and anti inflammatory activity (carrageenan-induced hind paw edema model) were used for all the extracts. In addition, the content of total phenolics, flavonoids and flavonols in all the extracts were determined with spectrophotometric methods. Results were compared with reference antioxidants via ascorbic acid, butylated hydroxytoluene, and gallic acid. Qualitative and quantitative compositions of all the extracts were analysed using a HPLC-PDA system. Polar fractions were found to be rich in flavonoids such as isovitexin and isoorientin. PMID- 17125947 TI - Interstitial deletion of a proximal 3p: a clinically recognisable syndrome. AB - Interstitial deletions of the proximal short arm of chromosome 3 occurring as constitutional aberrations are rare and a defined clinical phenotype is not established yet. We report on a 30-months-old girl with distinct facial features (square facies, plagiocephaly, broad forehead, broad nasal bridge, long philtrum and low set ears) and psychomotor/speech delay associated with an interstitial deletion of 3p12 chromosomal band, del(3)(p12p12). Clinical manifestations of our child were compared with those of other eight patients with the same deletion previously described to further delineate the proximal 3p deletion syndrome. PMID- 17125948 TI - Treatment of chronic inflammatory diseases with biologic agents: opportunities and risks for the elderly. AB - The treatment armamentarium in rheumatic inflammatory diseases has drastically increased in the last years. Earlier uses of conventional disease-modifying antirheumatic drugs (DMARDs), along with the arrival of newer therapies including the so-called "biologic" agents, have provided better long-term outcomes for patients suffering from these illnesses. Biologic agents have shown efficacy for several diseases and failed in others. Due to a high prevalence of some of these diseases in the elderly population, this age group may also benefit, although treatment will have to be tailored to its special needs. In this mini review, we will discuss the use of these medications in rheumatic diseases with a significant prevalence in the elderly, their proven and potential uses, and the considerations that need to be taken into account when using them in this population. PMID- 17125949 TI - Calorie restriction: progress during mid-2005-mid-2006. PMID- 17125950 TI - How vaccines work on the background of the aging immune system. AB - The elderly exhibit reduced responses to vaccinations, leaving them more susceptible to preventable infectious diseases such as influenza and pneumonia. It has been unclear as to the specific changes in the aging immune system that contribute to this decline, thus we have developed a mouse model to examine this phenomenon and determine why it occurs and how it can be overcome. Our ultimate goal is to determine how to enhance vaccine efficacy for elderly populations. PMID- 17125951 TI - Counting calories in Drosophila diet restriction. AB - The extension of life span by diet restriction in Drosophila has been argued to occur without limiting calories. Here we directly measure the calories assimilated by flies when maintained on full- and restricted-diets. We find that caloric intake is reduced on all diets that extend life span. Flies on low-yeast diet are long-lived and consume about half the calories of flies on high-yeast diets, regardless of the energetic content of the diet itself. Since caloric intake correlates with yeast concentration and thus with the intake of every metabolite in this dietary component, it is premature to conclude for Drosophila that calories do not explain extension of life span. PMID- 17125952 TI - Identification and characterization of degradation products of dicloxacillin in bulk drug and pharmaceutical dosage forms. AB - Impurity profiling of dicloxacillin sodium bulk drug and pharmaceutical dosage forms subjected to stability studies is evaluated. Of many impurities detected in HPLC analysis, three were not reported in the literature. The impurities have been identified by LC-MS; isolated by preparative HPLC; and characterised by NMR, Mass spectroscopy and IR. Pure impurities obtained by isolation were co-injected with dicloxacillin sodium sample to confirm the retention times in HPLC. Structure elucidation of these degradation products by spectral data has been discussed in detail. PMID- 17125953 TI - Quality control Fourier transform infrared determination of diazepam in pharmaceuticals. AB - A quality control procedure has been developed for the determination of diazepam in pharmaceuticals using Fourier transform infrared (FTIR) spectroscopy. The method involves the off-line extraction of diazepam with chloroform by sonication and direct determination in the extracts through peak area measurement in the interval between 1672 and 1682 cm(-1) using a baseline correction defined between 1850 and 1524 cm(-1). For standardization it was used an external calibration line established from standard solutions of diazepam in chloroform. The method provides a limit of detection of 0.04 mg per tablet (n=5), a relative standard deviation (R.S.D.) of 0.5% for 5 independent measurements of a standard solution at a concentration level of 0.76 mg g(-1) and a sampling frequency of the whole procedure of 4 h(-1), being required only 45 s for the measurement step. Results obtained by FTIR agree with those obtained by a reference methodology based on ultraviolet spectrometry and thus the developed procedure offers a good alternative for the determination of diazepam in pharmaceuticals. PMID- 17125954 TI - A liquid chromatography/mass spectrometric method for simultaneous analysis of arachidonic acid and its endogenous eicosanoid metabolites prostaglandins, dihydroxyeicosatrienoic acids, hydroxyeicosatetraenoic acids, and epoxyeicosatrienoic acids in rat brain tissue. AB - A sensitive, specific, and robust liquid chromatography/mass spectrometric (LC/MS) method was developed and validated that allows simultaneous analysis of arachidonic acid (AA) and its cyclooxygenase, cytochrome P450, and lipoxygenase pathway metabolites prostaglandins (PGs), dihydroxyeicosatrienoic acids (DiHETrEs), hydroxyeicosatetraenoic acids (HETEs) and epoxyeicosatrienoic acids (EETs), including PGF(2alpha), PGE(2), PGD(2), PGJ(2), 14,15-DiHETrE, 11,12 DiHETrE, 8,9-DiHETrE, 5,6-DiHETrE, 20-HETE, 15-HETE, 12-HETE, 9-HETE, 8-HETE, 5 HETE, 14,15-EET, 11,12-EET, 8,9-EET, and 5,6-EET in rat brain tissues. Deuterium labeled PGF(2alpha)-d(4), PGD(2)-d(4), 15(S)-HETE-d(8), 14,15-EET-d(8), 11,12-EET d(8), 8,9-EET-d(8), and AA-d(8) were used as internal standards. Solid phase extraction was used for sample preparation. A gradient LC/MS method using a C18 column and electrospray ionization source under negative ion mode was optimized for the best sensitivity and separation within 35 min. The method validation, including LC/MS instrument qualification, specificity, calibration model, accuracy, precision (without brain matrix and with brain matrix), and extraction efficiency were performed. The linear ranges of the calibration curves were 2 1000 pg for PGs, DiHETrEs, HETEs, and EETs, 10-2400 pg for PGE(2) and PGD(2), and 20-2000 ng for AA, respectively. PMID- 17125956 TI - "Patient-time", "doctor-time", and "institution-time": perceptions and definitions of time among doctors who become patients. AB - OBJECTIVE: To examine views and experiences of conflicts concerning time in healthcare, from the perspective of physicians who have become patients. METHODS: We conducted two in-depth semi-structured 2-h interviews concerning experiences of being health care workers, and becoming a patient, with each of 50 doctors who had serious illnesses. RESULTS: These doctor-patients often came to realize as they had not before how patients experience time differently, and how "patient time", "doctor-time", and "institution-time" exist and can conflict. Differences arose in both the long and short term, regarding historical time (prior eras/decades in medicine), prognosis (months/years), scheduling delays (days/weeks), daily medical events and tasks (hours), and periods in waiting rooms (minutes/hours). Definitions of periods of time (e.g., "fast", "slow", "plenty", and "soon") also varied widely, and could clash. Professional socialization had heretofore impeded awareness of these differences. Physicians tried to address these conflicts in several ways (e.g., trying to provide test results more promptly), though full resolution remained difficult. CONCLUSIONS: Doctors who became patients often now realized how physicians and patients differ in subjective experiences of time. Medical education and research have not adequately considered these issues, which can affect patient satisfaction, doctor patient relationships and communication, and care. PRACTICE IMPLICATIONS: Physicians need to be more sensitive to how their definitions, perceptions, and experiences concerning time can differ from those of patients. PMID- 17125957 TI - The effect of patient-centered contraceptive counseling in women who undergo a voluntary termination of pregnancy. AB - OBJECTIVE: The aim of the study was to evaluate, by means of a randomized controlled trial, whether a patient-centered contraceptive counseling intervention increased the use of contraception, and the knowledge and positive attitudes towards contraception, in women who undergo a termination of pregnancy (TOP). METHODS: The study was carried out at the San Paolo Hospital of Milan between the 1st of February and the 31st of May 2004. Participants (41 women; ages 20-44 years) were randomly divided into two groups: an experimental group (n = 20), who received patient-centered contraceptive counseling, and a control group (n = 21), who received the routine treatment in use at the San Paolo Hospital and were referred to the community health centers after the TOP. Both groups were administered a questionnaire at two points in time (before the counseling and 1 month later) which evaluated participants' knowledge, attitudes and use of contraception (the latter was also followed up 3 months later). The counseling intervention lasted 30 min and was carried out by a psychologist and a gynaecologist. RESULTS: It was found that knowledge, favorable attitudes and use of effective contraception increased significantly for the experimental group, whereas there was no significant change for the control group. CONCLUSION: The counseling intervention was therefore found to be efficacious in improving understanding and use of contraception in women who have undergone a TOP. The hope is that this will contribute to increased use of effective contraception in the future. PRACTICE IMPLICATIONS: Following the principles of patient-centered medicine, this study provides evidence for the importance of exploring woman's feelings, beliefs, wishes and expectations regarding contraception within a contraceptive counseling intervention. PMID- 17125958 TI - Influences on patients' ratings of physicians: Physicians demographics and personality. AB - OBJECTIVE: There is considerable interest in the influences on patients' ratings of physicians. METHODS: In this cross-sectional study, patients (n = 4616; age range: 18-65 years) rated their level of satisfaction with their primary care physicians (n = 96). Patients and physicians were recruited from primary care practices in the Rochester, NY metropolitan area. For analytic purposes, length of the patient-physician relationship was stratified (< or =1, 1-4, > or =5 years). Principal components factor analysis of items from the Health Care Climate Questionnaire, the Primary Care Assessment Survey and the Patient Satisfaction Questionnaire yielded a single factor labeled "Satisfaction" that served as the sole dependent variable. Higher scores mean greater satisfaction. Predictors of interest were patient demographics and morbidity as well as physician demographics and personality, assessed with items from the NEO-FFI. RESULTS: Patients treated by a physician for 1 year or less rated male physicians higher than female physicians. This gender difference disappeared after 1 year, but two physician personality traits, Openness and Conscientiousness, were associated with patients' ratings in lengthier patient-physician relationships. Patients report being more satisfied with physicians who are relatively high in Openness and average in Conscientiousness. Older patients provide higher ratings than younger patients, and those with greater medical burden rated their physicians higher. CONCLUSION: Patients' ratings of physicians are multidetermined. Future research on patient satisfaction and the doctor-patient relationship would benefit from a consideration of physician personality. Identifying physician personality traits that facilitate or undermine communication, trust, patient-centeredness, and patient adherence to prescribed treatments is an important priority. PRACTICE IMPLICATIONS: Learning environments could be created to reinforce certain traits and corresponding habits of mind that enhance patient satisfaction. Such a shift in the culture of medical education and practice could have implications for patient care. PMID- 17125959 TI - Surgeon volume and postoperative mortality after oesophagectomy for cancer. AB - AIM: Oesophagectomy remains the curative treatment of choice for patients with localised oesophageal or cardia cancer, but severe postoperative complications are common. Our aim was to assess the association between surgeon volume and postoperative mortality after oesophagectomy. METHODS: Prospective, population based study of Swedish residents diagnosed with oesophageal or cardia cancer, treated with oesophagectomy during the period April 2001 through December 2005. Details concerning patients, tumours, and surgery were collected from the Swedish Oesophageal and Cardia Cancer register. All 607 patients registered during the study period were included in the study. Risk of mortality 30 and 90 days after oesophagectomy was assessed using multivariable logistic regression, expressed in odds ratios (OR) with 95% confidence intervals (CI), adjusted for relevant covariates. RESULTS: The 30-day mortality in low-, medium-, and high-volume surgeon groups were 7.1%, 2.1%, and 2.6%, respectively. The corresponding 90-day figures were 11.4%, 4.8%, and 8.9%. Adjusted ORs for 30- and 90-day mortality were decreased non-significantly by 58% and 14%, respectively, among patients in the high-volume group, compared to the low-volume group (OR 0.42, 95% CI 0.10 1.80; OR 0.86, 95% CI 0.31-2.38). The mortality rates differed considerably between individual high-volume surgeons, but without any trend of further decreased risk with increasing volume among these surgeons (p values for trend 0.84 and 0.80 for 30- and 90-day mortality, respectively). CONCLUSION: Patients with resectable oesophageal cancer should be advised to choose a high-volume surgeon, but they should also be aware that differences among individual surgeons might further affect survival. PMID- 17125960 TI - Natural history, diagnosis, treatment and outcome of medullary thyroid cancer: 37 years experience on 157 patients. AB - AIM: The analysis of a 37-year retrospective study on diagnosis, prognostic variables, treatment and outcome of a large group of medullary thyroid cancer (MTC) patients was conducted, in order to plan a possible evidence-based management process. METHODS: Between Jan 1967 to Dec 2004, 157 consecutive MTC patients underwent surgery in our centre: 60 males and 97 females, mean age 47.3 years (range 6-79). Total thyroidectomy was performed in 143 patients (91.1%); central compartment (CC) node dissection (level VI) in 41 patients; central plus lateral compartment (LC) node dissection (levels II, III, and IV) in 82 patients. Subtotal thyroidectomy was initially performed in 14 cases: 10 of them were re operated because of persistence of elevated serum calcitonin levels. RESULTS: After a median post-surgical follow-up of 68 months (range 2-440 months), 42.9% of patients were living disease-free, 39.8% were living with disease, 3.1% were deceased due to causes different from MTC, and 3.2% were deceased due to MTC. The overall 10-year survival rate was 72%. At uni-variate statistical analysis (a) patient's age at initial treatment (>45 years; >/=45 years), (b) sporadic vs. hereditary MTC, (c) disease stage, and (d) the extent of surgical approach resulted as significant variables. Instead, at multivariate statistical analysis, only (a) patient's age at initial diagnosis, (b) disease stage, and (c) the extent of surgery resulted as significant and independent prognostic variables influencing survival. CONCLUSION: The presence of lymph node and distant metastases at first diagnosis significantly worsened prognosis and survival rate in our series. Early diagnosis of MTC is very important, allowing complete surgical cure in Stages I and II patients. Due to the relatively bad prognosis of MTC, especially for disease Stages III and IV, it appears reasonable to recommend radical surgery including total thyroidectomy plus CC lymphoadenectomy as the treatment of choice, plus LC lymphoadenectomy in patients with palpable and/or ultrasound enlarged neck lymph nodes. PMID- 17125961 TI - Overexpression of Cripto and its prognostic significance in breast cancer: a study with long-term survival. AB - INTRODUCTION: Cripto is a founding member of the EGF-CFC family, and plays an important role in tumourigenesis, tumour cell proliferation and migration. We aimed to determine the significance of Cripto expression on the survival of patients with breast cancer. METHODS: Immunohistochemical detection of Cripto was performed by using mAb C13 on 120 formalin-fixed paraffin-embedded breast tumour specimens in tissue microarrays. This cohort comprises a series of 120 patients with primary operable breast cancer diagnosed between 1989 and 1995, retrieved from the Concord Repatriation General Hospital breast carcinoma database. RESULTS: Using a cutoff value of 80%, Cripto overexpressed in 57 of the 120 (47.5%) patients. We found significant associations between overexpression of Cripto and the Nottingham Prognostic Index (NPI, p<0.01), histological grade (p<0.01), pathological tumour type (p=0.04), PR (p=0.02) as well as Ki-67 (p=0.02). Univariate analysis reveals that there is a significant correlation between overexpression of Cripto and survival (p=0.0003). Cox regression analysis indicates that the overexpression of Cripto is an independent prognostic factor in breast cancer (HR 2.79, 95%CI 1.20-6.50). CONCLUSION: The unique epitope recognized by mAb C13 is overexpressed on breast tumour tissues. In this series of invasive breast cancers, overexpression of Cripto was more often found in high grade and poor prognosis tumours compared to low grade and good prognosis breast cancers. Moreover, overexpression of Cripto was significantly associated with decreased patient survival. PMID- 17125962 TI - Immediate maxillary reconstruction after malignant tumor extirpation. AB - AIMS: Immediate maxillary reconstruction after malignant tumor extirpation differs from other types of maxillary reconstruction. Our reconstruction algorithm is described in this article. METHODS: One hundred ninety-four patients who had undergone maxillectomy for malignant tumors were reviewed, and maxillectomy defects were classified with the method of Cordeiro and Santamaria. RESULTS: Mean total blood loss was 848 ml, and 71 patients died within 2 years after surgery. For type IIIa defects of the orbital floor, titanium mesh or vascularized bone or cartilage was used for reconstruction, but the rate of postoperative complications did not differ between titanium and autografts. Therefore, to reconstruct orbital floor defects we have recently used only titanium mesh. For type I or II defects, we use autografts for only selected cases. CONCLUSIONS: We strive to perform less-invasive reconstructive surgery after resection for maxillary malignancy. PMID- 17125963 TI - Predictors of axillary lymph node metastasis in breast cancer: a systematic review. AB - AIMS: To review the established and emerging techniques in axillary lymph node prediction and explore their potential impact on clinical practice. To reliably identify patients in whom axillary lymph node surgery, including SLNB, can be safely omitted. METHODS: Searches of PubMed were made using the search terms "axilla" (or "axillary"), "lymph", "node" and "predictor" (or "prediction"). Articles from abstracts and reports from meetings were included only when they related directly to previously published work. FINDINGS: There are numerous studies in which the predictive utility of biomarkers as determinants of axillary lymph node status have been investigated. Few of these have specifically addressed the attributes of the primary tumour which could offer much potential for the prediction of tumour metastasis to the axillary lymph nodes. CONCLUSIONS: Currently, no single marker is sufficiently accurate to obviate the need for formal axillary staging using SLNB or axillary clearance. PMID- 17125964 TI - Bovine plasma proteins increase virulence of Haemophilus somnus in mice. AB - The role of bovine serum or plasma proteins in Haemophilus somnus virulence was investigated in a mouse model of septicemia. An increase in virulence was detected when the organism was pre-incubated for 5 min and inoculated with fetal calf serum. When purified bovine serum or plasma proteins were pre-incubated with H. somnus before inoculating into mice, transferrin was found to increase virulence. Bovine lactoferrin was also noted to increase virulence, but to a lesser extent and had a delayed time course when compared with transferrin. Using an ELISA assay, an increased amount of H. somnus whole cells and culture supernatant bound to bovine transferrin when the organism was grown in iron restricted media. Lactoferrin also bound to H. somnus, but binding was not affected by growth in iron-restricted media and it was eliminated with 2M NaCl, which reversed charge mediated binding. Transferrin, but not lactoferrin, supported growth of H. somnus on iron-depleted agar based media using a disk assay. Therefore, lactoferrin increased virulence by an undetermined mechanism whereas transferrin increased virulence of H. somnus by binding to iron-regulated outer-membrane proteins (IROMPs) and providing iron to the pathogen. PMID- 17125965 TI - In vitro toxicity evaluation of single walled carbon nanotubes on human A549 lung cells. AB - This paper describes the in vitro cytotoxicity assessment of single walled carbon nanotubes (SWCNT) on A549 cells, a human lung cell line. Cellular viability was determined using the alamar blue (AB), neutral red (NR) and MTT assays, which evaluated metabolic, lysosomal and mitochondrial activity respectively. In addition, the total protein content of the cells was measured using the coomassie brilliant (CB) blue assay. Supernatants were also assayed for Adenylate Kinase (AK) release and Interleukin 8 (IL-8) which indicated a loss of cell membrane integrity and an inflammation response respectively. To investigate the interactions between serum components in the test medium and the test materials, exposures were conducted both in serum containing (5%) and serum-free medium. Results from the cytotoxicity tests (AB, CB, MTT) revealed the SWCNT to have very low acute toxicity to the A549 cells as all but one of the reported 24h EC(50) values exceeded the top concentration tested (800 microg/ml). The SWCNT were found to interfere with a number of the dyes used in the cytotoxicity assessment and we are currently conducting a comprehensive spectroscopic study to further investigate these interactions. Of the multiple cytotoxicity assays used, the AB assay was found to be the most sensitive and reproducible. Transmission electron microscopy (TEM) studies confirmed that there was no intracellular localization of SWCNT in A549 cells following 24h exposure; however, increased numbers of surfactant storing lamellar bodies were observed in exposed cells. PMID- 17125966 TI - Manipulating perceptions of spider characteristics and predicted spider fear: evidence for the cognitive vulnerability model of the etiology of fear. AB - The present study reports on an attempt to experimentally manipulate perceptions of uncontrollability, unpredictability and dangerousness related to an imaginal encounter with a spider in order to determine whether there is an effect on self rated predicted spider fear. Experimental manipulations involved differing information in relation to both the spider and the imaginal task. The control, predictability and dangerousness manipulations all had significant main effects on task-related spider fear (TRSF). Measures of the perception of the spiders as uncontrollable, unpredictable and dangerous were also significantly associated with TRSF and accounted for 42% of the variance in predicted fear beyond that accounted for by the experimental manipulations. Results are discussed in terms of their implications for better understanding the etiology and maintenance of fear. The overall findings are consistent with the cognitive vulnerability model, with cognitive perceptions of an object or situation seen as causal determinants of the fear associated with the stimulus. PMID- 17125967 TI - Evaluating microarrays using a semiparametric approach: application to the central carbon metabolism of Escherichia coli BL21 and JM109. AB - Escherichia coli K (JM109) and E. coli B (BL21) are strains used routinely for recombinant protein production. These two strains grow and respond differently to environmental factors such as glucose and oxygen concentration. The differences have been attributed to differential expression of individual genes that constitute certain metabolic pathways that are part of the central carbon metabolism. By implementing a semiparametric algorithm, which is based on a density ratio model, it was possible to compare and quantify the expression patterns of groups of genes involved in several central carbon metabolic pathways. The groups comprising the glyoxylate shunt, TCA cycle, fatty acid, and gluconeogenesis and anaplerotic pathways were expressed differently between the two strains, whereas no differences were apparent for the groups comprising either glycolysis or the pentose phosphate pathway. These results further characterized differences between the two E. coli strains and illustrated the potency of the semiparametric algorithm. PMID- 17125968 TI - Reversible periictal MRI abnormalities: clinical correlates and long-term outcome in 12 patients. AB - Although a wide spectrum of reversible periictal magnetic resonance imaging (MRI) abnormalities (RPMA) are being increasingly identified, the clinicians are often in a dilemma about their localization significance. This prompted us to analyze the clinical, MRI, electroencephalographic (EEG) and follow-up data of 12 patients with RPMA seen in a tertiary referral epilepsy center. RPMA occurred after a single or a cluster of focal seizures with or without secondary generalization. The interictal and ictal EEG abnormalities were localized to the site of RPMA in nine patients. RPMA involved areas remote from the site of EEG abnormalities in four patients. We have developed a comprehensive classification to account for the wide spectrum of RPMA involving gray matter, white matter and leptomeninges with or without contrast enhancement or mass effect. Follow-up MRIs showed complete resolution of RPMA in all, except in four patients, who developed residual focal atrophy. During median follow-up period of 3 years, recurrence of RPMA was observed in two patients. Diffusion weighted MRI in two patients and histopathological finding in one patient favored causal role of hypoxia in the pathogenesis of RPMA. Our observations help to understand the electroclinical profile, radiological spectrum, localization significance and natural history of RPMA better. PMID- 17125969 TI - The controversial association of ABCB1 polymorphisms in refractory epilepsy: an analysis of multiple SNPs in an Irish population. AB - Controversy has surrounded the reported association of the single nucleotide polymorphism (SNP) C3435T of the ATP-binding cassette subfamily B member 1 (ABCB1, MDR1) gene, with refractory epilepsy. Here we examine this question by: (1) attempting to replicate the original association, (2) assessing the association of other variants in high linkage disequilibrium (LD) with C3435T, and (3) evaluating and comparing our findings with other published studies. We defined drug-responsiveness as seizure freedom or a 50% or more reduction in seizure frequency in the preceding year. Drug resistance was defined as a less than 50% reduction in seizure frequency in the preceding year. We used a combination of map-based (tagging SNPs) and direct sequence-based methods allowing a comprehensive assessment of variation across the associated interval. Genotypic data on 8 SNPs was collected on 440 patients, of whom 242 were drug responsive and 198 were drug-resistant. We were unable to observe the original association of drug-resistant epilepsy with C3435T, nor any association with other functional variants at SNP or haplotype level in the ABCB1 gene. Evaluation of other attempted replication studies suggest that if the original C3435T association is indeed real, it would appear highly sensitive to the phenotype used. Alternatively, the discrepant results of this and other association attempts may be indicative of the original report of the CC genotype at C3435T in ABCB1 being a false positive finding. Variability in phenotypic descriptions in genetic association studies may partly explain the inconsistency of attempted replications. PMID- 17125970 TI - Extensive linkage disequilibrium mapping at HTR2A and DRD3 for schizophrenia susceptibility genes in the Galician population. AB - The serotonin and dopamine neurotransmitter systems are candidate pathways in the development of schizophrenia because of the assumed causal relationship with the observed symptoms as well as effective targeting of the corresponding receptors by antipsychotic drugs. However, genetic association studies have systematically focused on a limited set of genes and single nucleotide polymorphisms (SNPs), including T102C at HTR2A and Ser9Gly at DRD3. Meta-analyses of the associations between these two markers and schizophrenia revealed a true increase in risk, the magnitude of the effect being very low. In the present study we analyzed 260 schizophrenic patients and 354 control subjects from a homogeneous population, the Galician population, using an extensive linkage disequilibrium (LD) mapping approach, genotyping a total of 47 SNPs to test for the existence of additional variants that confer higher risk. We detected nominal significant association with schizophrenia for several haplotype tag SNPs (htSNPs) at HTR2A, although the significance was lost after multiple test corrections. In addition, haplotype analyses involving a sliding window approach, with window size 2 to 4 SNPs, revealed significant differences in frequencies of the DRD3 haplotypes at the 3' half of the gene region. This difference, which remains clearly significant after multiple test corrections (p=0.002, 0.0001, and 0.0025, for window sizes 2, 3, and 4, respectively), was mainly due to over-representation of several rare haplotypes in patients, at the expense of a single common haplotype; this represents interesting evidence of rare haplotypes for susceptibility detected using common htSNPs due to their strong effect. PMID- 17125971 TI - paa, originally identified in attaching and effacing Escherichia coli, is also associated with enterotoxigenic E. coli. AB - Previous studies on virotypes and antimicrobial resistance in a collection of porcine enterotoxigenic Escherichia coli (ETEC) O149 strains from Quebec revealed an increase in the number of multiresistant strains (in particular to tetracycline) and the appearance of new virulence factors with time. Among these factors is paa (for porcine attaching- and effacing-associated), originally identified in a porcine enteropathogenic strain, but also present in enterohemorrhagic E. coli O157:H7. In the present study, the association of paa with other ETEC virulence genes, its conservation and expression were investigated in the O149 ETEC collection. All 37 paa-positive strains possessed estB, elt, astA and faeG, and more than half also carried the estA gene, defining two main virotypes, estA(+) and estA(-). Most strains were tetA- or tetB positive, or both. paa is carried on high molecular weight plasmids. On tetA plasmids, paa is mostly found with enterotoxin gene estA and autotransporter gene sepA. Paa, a 30 kDa protein, is highly conserved and expressed in these strains. Moreover, paaETEC and porcine EPEC/EHEC contain IS signatures, suggesting that paa could be derived from a common ancestor. All these observations suggest a broader role than previously assessed in virulence for paa. PMID- 17125972 TI - A Bacteroides fragilis surface glycoprotein mediates the interaction between the bacterium and the extracellular matrix component laminin-1. AB - The adherence of Bacteroides fragilis strains to immobilized laminin-1 (LMN-1) was investigated using this protein adsorbed onto glass. Among the 27 strains isolated from infectious processes and assayed, 13 presented strong adherence to LMN-1. Among them, two strains, MC2 and 1081, showed the strongest association, and for that reason they were selected for further studies in which adherence to this protein was confronted with both physical-chemical and enzymatic treatments, along with concurrence assays with the LMN-1 molecule itself and the LMN-1 residing amino acid sequences (RGD, IKVAV, YIGSR, AG73, A13 and C16). The chemical and enzymatic treatments resulted in sharp decreases in binding rates of those strains, and competition experiments with LMN-1- residing amino acids revealed that, except for RGD and A13, all the others were effective at reducing bacterial binding of the bacteria. The outer membrane proteins (OMPs) of B. fragilis were extracted and assayed onto dot-blotted LMN-1, and when the extracts were chemically treated, especially with metasodium periodate, a drastic reduction in bacterial binding occurred. Results of the latter assays clearly indicate that bacterial molecules involved in both recognition and binding of B. fragilis to LMN-1 are present in OMP extracts. Taken together, our results strongly indicate that a B. fragilis surface glycoprotein may play a key role in bacterial association with LMN-1. PMID- 17125973 TI - Quantification of bacterial mass recovery as a function of pore-water velocity and ionic strength. AB - Transport of bacteria in aquifer systems plays an important role in bioaugmentation, which relies upon successful bacterial delivery to a target area. In the present study, we conducted a set of laboratory column experiments under various conditions of pore-water velocity (upsilon(omega)) and ionic strength (IS) of culture medium for Pseudomonas aeruginosa, known to be a benzene degrading bacteria, in order to investigate their relationship to mass recovery in saturated quartz sands. The column experiments revealed that both peak concentrations and mass recoveries of bacteria were lower than those of a conservative tracer KCl when deionized water was used as leaching water for all ranges of pore-water velocity (0.18-6.23 cm/min). Thus, the parameter responsible for transport of P. aeruginosa was only the deposition coefficient. Bacterial cells could not be attached to the mineral surfaces by predominance of electrostatic charge or repulsive forces over hydrophobicity or attractive forces due to the very low ionic strength ( approximately 0 mM) of deionized water. The loss of bacterial mass was attributed to the deposition in the crevice formed on the quartz surfaces, as evidenced by SEM images. For a given pore-water velocity, the ionic strength markedly influenced bacterial deposition, showing decreased peak concentrations and mass recoveries with increasing ionic strength of column leaching water. An optimum range of upsilon(omega) and IS for achieving bacterial mass recovery higher than 70% in the studied quartz sand was found such that: (i) at low IS ( approximately 0 mM), a pore-water velocity higher than 0.30 cm/min, and (ii) at pore-water velocity of 0.52 cm/min, an IS lower than 290 mM, were required, respectively. PMID- 17125974 TI - Comparison of amplified ribosomal DNA restriction analysis, peptidoglycan hydrolase and biochemical profiles for rapid dairy propionibacteria species identification. AB - Species of dairy propionibacteria are used as cheese-ripening cultures as well as probiotics. However, no rapid identification methods are currently available. With this in mind, the present study compared three methods, (i) carbohydrate fermentation, (ii) ARDRA (amplified ribosomal DNA restriction analysis) and (iii) peptidoglycan hydrolase (PGH) activity profiles to improve the identification of Propionibacterium thoenii, Propionibacterium jensenii, Propionibacterium acidipropionici and Propionibacterium microaerophilum. The species Propionibacterium freudenreichii and Propionibacterium cyclohexanicum have previously been shown to be easily distinguishable from the other species. Principal component analysis of the carbohydrate fermentation profiles of 113 P. thoenii, P. jensenii, P. acidipropionici and P. microaerophilum strains correctly classified 85% of the strains based on the fermentation of seven carbohydrates. Regarding PGH profiles, optimized conditions of PGH-renaturing SDS-PAGE were applied to 34 of the strains. The PGH profiles of P. acidipropionici and P. microaerophilum were indistinguishable from one another, but were easily distinguished from P. jensenii and P. thoenii. However, four strains exhibited atypical profiles. Hence, in general, the PGH profiles were shown to be conserved within a species, with some exceptions. Four endonucleases were tested for ARDRA and the four species differentiated by combining the profiles obtained with MspI and HaeIII. P. freudenreichii and P. cyclohexanicum profiles were also performed but showed wide differences. Consequently, ARDRA was shown to be the most appropriate method for rapidly distinguishing strains of propionibacteria. Carbohydrate fermentation and peptidoglycan hydrolase activity profiles are useful as complementary identification tools, since about 15% of the 34 strains tested showed atypical profiles. PMID- 17125975 TI - Occurrence and diversity of integrons and beta-lactamase genes among ampicillin resistant isolates from estuarine waters. AB - The aim of the present study was to assess the occurrence and molecular diversity of beta-lactamase genes and integrons among Gram-negative ampicillin-resistant bacteria from Ria de Aveiro. Ampicillin-resistant isolates were selected and subjected to genotyping using REP-PCR. Representatives from each REP pattern were affiliated with the following taxa by sequencing the 16S rRNA gene: Aeromonas caviae, A. hydrophila, A. media, A. molluscorum, A. veronii, A. salmonicida, Aeromonas sp., Pseudomonas putida, Pseudomonas sp., Escherichia coli, Escherichia sp., Shigella sonnei, Shigella sp., Klebsiella pneumoniae, K. oxytoca, Raoultella ornithinolytica, R. terrigena, R. planticola, Citrobacter freundii, Morganella morganii and Enterobacter sp. Isolates affiliated with genera Escherichia or Shigella were identified as Escherichia coli using phenotypic-based tests. PCR was used to assess beta-lactamase encoding sequences (bla(TEM), bla(SHV), bla(CARB), bla(CTX-M), bla(IMP), bla(VIM), bla(CphA/IMIS), bla(OXA-A), bla(OXA B), bla(OXA-C)), class 1 and class 2 integrases, and integron variable regions. Sequence analysis of PCR products was performed. beta-Lactamase genes were detected in 77.8% of the Enterobacteriaceae and in 10.5% of the Aeromonas. The most frequently detected gene was bla(TEM), followed by bla(SHV,)bla(OXA-B), bla(CphA/IMIS) and bla(CARB). Retrieved sequences shared high homology with previously described beta-lactamases. The intI1 gene was present in 29.6% of the Enterobacteriaceae and in 21% of the Aeromonas isolates. The intI2 gene was present in 4 isolates. A total of 13 cassettes included in 12 different cassette arrays were identified. The most frequently found resistance gene cassettes were aadA variants. Previous investigations based on cultivation-independent approaches revealed higher molecular diversity among beta-lactamase-encoding sequences in this estuary. This fact reinforces the hypothesis that cultivation dependent approaches may underestimate the prevalence of antibiotic resistance genes in environmental samples and may introduce bias in the recovery of their molecular variants. PMID- 17125976 TI - Silica xerogel-hydrogen peroxide composites: their morphology, stability, and antimicrobial activity. AB - Hydrogen peroxide was incorporated into silica xerogel matrix over the concentration range from 3.8 to 68.0 wt% via the sol-gel route. The obtained composites were characterized by scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). The release rates of H(2)O(2) from the composites into the aqueous phase were examined. In most cases, a 90% release was attained after ca. 10 min, and it was only slightly dependent on H(2)O(2) concentration and particle size. The antimicrobial activity of the composite containing 3.59% H(2)O(2) was evaluated against Escherichia coli and Micrococcus luteus. A comparative assay was carried out for aqueous solution of H(2)O(2) of the same concentration. The results demonstrated a potent microbicidal efficacy of the composite. Furthermore, diffusion range of the hydrogen peroxide from the solid composite into an agar medium matched that of the H(2)O(2) in aqueous solution. The stability tests with the xerogels containing 3.8, 26.4, and 68.0% of H(2)O(2) showed that after 63 days respective losses of the H(2)O(2) at 3 degrees C were 8.8, 9.7, and 6.2%. Both the DSC results and the stability tests have shown that the molecular water present in the pores stabilizes the composite, probably through improving the binding of the H(2)O(2) molecules onto the silica surface. PMID- 17125977 TI - Dynamic electro-optic properties of macromolecules and nanoparticles in solution: a review of computational and simulation methodologies. AB - This paper reviews some theories, and computational and simulation procedures available for the calculation of the time-course of electro-optic properties of particles in solution. For rigid particles, the time evolution of the properties is directly related to their rotational diffusion; therefore, the computational procedures for the calculation of hydrodynamic properties find a direct application in electro-optics. Several of such computational procedures, based on bead models, are reviewed. For flexible particles, the simultaneous effects the external field and the flexibility can be treated with Brownian dynamics simulation. We illustrate the various procedures, with applications to rigid bent rods and flexible, wormlike or hinged rods, trying to show how the absence or presence of flexibility, and its kind, influences the dynamic electro-optic properties, which are therefore valuable sources of information about the conformation of macromolecules and nanoparticles. PMID- 17125978 TI - Na+/Ca2+ exchanger inhibitors modify the accumulation of tumor-diagnostic PET tracers in cancer cells. AB - AIM: To establish the effects of Na(+)/Ca(2+) exchanger (NCX) blockers on 2 [(18)F]fluoro-2-deoxy-D-glucose ((18)FDG) and (11)C-choline accumulation in different cancer cells. METHODS: The tumor cells were incubated with NCX inhibitors, and the uptakes of (18)FDG and (11)C-choline were measured. Flow cytometric measurements of intracellular Ca(2+) and Na(+) concentrations were carried out. The presence of the NCX antigen in the cancer cells was proved by Western blotting, flow cytometry and confocal laser scanning microscopy. RESULTS: The NCX is expressed at a noteworthy level in the cytosol and on the cytoplasmic membrane of the examined cells. Incubation of the cells with three chemically unrelated NCX blockers (bepridil, KB-R7943 or 3',4'-dichlorobenzamil hydrochloride) resulted in an increase in the intracellular Ca(2+) concentration, with a simultaneous decrease in the intracellular Na(+) concentration. The treatment with the NCX inhibitors increased the energy consumption of the tumor cells by 50-100%. Thapsigargin abolished the NCX-induced (18)FDG accumulation in the cells. The NCX blockers applied decreased the (11)C-choline accumulation of all the investigated cancer cells by 60-80% relative to the control. CONCLUSION: A possible masking effect of NCX medication must be taken into consideration during the diagnostic interpretation of PET scans. PMID- 17125979 TI - [Thyroiditis and gluten intolerance: extrapancreatic auto-immune diseases associated with type 1 diabetes]. AB - OBJECTIVES: This study aimed at evaluating the screening of thyroiditis and coeliac disease, in a population of children and adolescents with type 1 diabetes, and at comparing the appearance of antibodies specific for these 2 diseases as a function of age. PATIENTS AND METHODS: The study included 370 children and adolescents, 179 girls and 191 boys, aged 13.8 +/- 4.4 yr and with diabetes for 7.1 +/- 3.8 yr. Auto-immune thyroiditis was screened using antimicrosomal and antithyroglobulin auto-antibodies, at a mean rhythm of 3 tests per patient (1 every 2 yr), associated with dosages of TSH and FT4. Coeliac disease was screened using antigliadin (+/- antiendomysium) auto-antibodies, at a mean rhythm of 2 tests per patient, and was confirmed by duodenojejunal biopsy. Antithyroid auto-antibodies were correlated with age following the "censured data analysis" type approach. RESULTS: Antithyroid autoantibodies were found in 42 patients (11.4%), of whom 9 were treated for hypothyroidism and 1 for Basedow disease, and coeliac disease autoantibodies were found in 9 patients (3.2% of tested patients). The cumulated frequency of antithyroid auto-antibodies increased regularly with age and was significantly higher in girls, reaching 28% in girls and 12% in boys around 18 yr of age. As a consequence of this evolution, antithyroid auto-antibodies were frequently found at the time of diagnosis of diabetes when it declared after 10 yr of age, while they often became positive secondarily when diabetes occurred before 10 yr of age. Coeliac disease specific auto-antibodies appeared much earlier and were found at the time of diagnosis of diabetes or at the first screening test. CONCLUSION: Antithyroid autoantibodies are increasingly frequent with age in children with type 1 diabetes, and become very elevated in girls. The rhythm for screening should be adapted to this evolution of autoantibodies with age, which is very different between thyroiditis and coeliac disease. PMID- 17125980 TI - [Management of intussusception in France in 2004: investigation of the Paediatric Infectious Diseases Group, the French Group of Paediatric Emergency and Reanimation, and the French Society of Paediatric Surgery]. AB - OBJECTIVES: To describe the different pathways of management of intussusception (IS) in infants and children in metropolitan France and to identify paediatric emergency centres that might constitute a surveillance network for IS. MATERIAL AND METHODS: A questionnaire was sent to 273 paediatric emergency centres distributed across France in 2005. Modalities of diagnosis and treatment of IS had to be precised. RESULTS: One hundred and sixty-seven centres (61.2%) responded. The response was given by 131 paediatricians (78.4%) and 36 surgeons (21.6%) working in 38 universitary hospitals (22.7%) and 129 general hospitals (77.2%). The mean number of IS treated in each centre in 2004 was 11+/-13.5 (extr. 0 to 70; median 6). Diagnosis of IS required a collaboration between medical and surgical teams in 51.5% of the centres, but in 40.1% the sole medical team was in charge of the diagnosis. Ultrasonography is used for diagnosis by 98.8% of the centres. Reduction with hydrostatic enema and eventually surgery was performed in the same hospital in 44.3%. Other centres systematically or frequently transferred the patients for reduction, mostly towards universitary hospitals (90%). CONCLUSION: The procedures of IS diagnosis are the same everywhere in France but the pathways of therapeutic management do vary, depending on the availability of surgeons and anaesthetists trained in paediatrics on each site. These disparities will probably change with the implementation of the new plan for sanitary organization in children and adolescents in France. Labellized paediatric emergency centres will gather more surgical patients and could eventually constitute an effective surveillance network for IS. PMID- 17125981 TI - Redescriptions of two marine planktonic ciliates from China, Parastrombidium faurei (Kahl, 1932) Maeda, 1986 and Strombidium capitatum (Leegaard, 1915) Kahl, 1932 (Ciliophora, Oligotrichea). AB - The morphology and infraciliature of two marine planktonic ciliates, Parastrombidium faurei (Kahl, 1932) Maeda, 1986 and Strombidium capitatum (Leegaard, 1915) Kahl, 1932, from coastal waters near Qingdao (Tsingtao), northern China, were investigated based on live observation and protargol impregnation. This is the first time that these two species are reported from Yellow Sea/North Pacific. Emended diagnoses for the genus Parastrombidium and its single species P. faurei are provided as they were not studied with modern methods before. The ciliary pattern and the stomatogenesis of Parastrombidium indicate that it belongs to the order Choreotrichida Small and Lynn, 1985, family Strombidinopsidae Small and Lynn, 1985. An updated description for the "well known"Strombidium capitatum is also included based on the Qingdao population. PMID- 17125982 TI - Development of spherical iron(II) sulfate heptahydrate-containing solid particles with sustained drug release. AB - The aim of this work was to develop a simple, economic procedure for the manufacturing of coated iron(II) sulfate particles by using a crystallization technique for the development of round particles, followed by coating with a lipophilic material. Several batches of iron(II) sulfate heptahydrate were produced by a cooling crystallization, with variation of the crystallization parameters. The spherical crystals were coated with stearin. The products were characterized for particle size, roundness, bulk density and in vitro drug dissolution. Crystallization was performed from deionized water with no addition of seed crystals and by cooling by applying a linear cooling rate. The developed iron(II) sulfate crystals were round with average diameter of 729+/-165 microm. The best form for the sustained release of iron(II) sulfate was the sample HTP-2 which contained 11% of stearin relative to the iron(II) sulfate. The spherical crystallization of iron(II) sulfate is simple and fast, and does not require a dangerous, expensive solvent. The round particles can coat directly with lipophilic material which results in slow release of iron(II) sulfate and protects the iron(II) from oxidation and inhibits the loss of crystal water. The coated crystals can be filled into capsules to yield the final dosage form. PMID- 17125983 TI - Determinants of variability of protein content, volume and pH of exhaled breath condensate. AB - Collection of exhaled breath condensate (EBC) is a simple and noninvasive method to obtain information on the respiratory system. Different mediators can be determined in EBC. However, determinants of variability are not well described. The aim of this study was to evaluate variability of pH, volume and protein concentration of EBC between individuals and between sampling times. Therefore, EBC was collected from 20 healthy volunteers on two different days. Median pH for all samples, measured 5 min after collection without deaeration, was 6.17. Median volume was 1.70 ml and median total protein concentration was 1.02 microg/ml. Coefficients of variation were 5.17%, 21.84% and 37.93%, respectively. No intra- or interday variability could be found, except for the first collection time. Between individuals, significant differences were observed for all three mediators. Age, height and gender can explain part of this variation. In conclusion, no significant difference between sampling times on the same day or on different days was obtained for pH, volume and total protein concentration, provided that subjects are experienced in collecting EBC. PMID- 17125984 TI - Inhaled budesonide in the management of acute worsenings and exacerbations of asthma: a review of the evidence. AB - The use of systemic corticosteroids, together with bronchodilators and oxygen therapy, has become established for the management of acute asthma. These agents are undoubtedly effective, but are also associated with problems such as metabolic adverse effects. Inhaled corticosteroids (ICS) offer potential benefit in the acute setting because they are delivered directly to the airways. They are also likely to reduce systemic exposure, which would lead in turn to reductions in rates of unwanted systemic effects. In order to evaluate the role of budesonide in the management of acute asthma exacerbations we conducted a review of the literature and critically evaluated the rationale for the use of ICS in general in this setting. Trials in adults and children requiring treatment for acute exacerbation of asthma have shown clinical and/or spirometric benefit for budesonide when delivered via nebulizer, dry powder inhaler, or aerosol in the emergency department, hospital and follow-up settings. The efficacy seems to benefit from high doses given repeatedly during the initial phase of an acute exacerbation. These acute effects are likely to be linked to the drug's distinctive pharmacokinetic and pharmacodynamic profile. The current evidence base revealed encouraging results regarding the efficacy of the ICS budesonide in patients with wheeze and acute worsening of asthma. Future studies should focus on the efficacy of these agents in more severe asthma worsenings. PMID- 17125985 TI - Removal of manganese and zinc from Kahrizak landfill leachate using daily cover soil and lime. AB - The possibility of in situ removal of heavy metals found in leachate generated at municipal solid waste landfills was studied through amendment of daily cover soil. Kahrizak landfill, which receives the waste generated at Tehran, was selected as the source of leachate and soil samples. Manganese and zinc were selected in this study. The soil sample taken from the Kahrizak site contained about 17% clay, which was presumed to have significant capability for removing manganese and zinc. This capability was assumed to be enhanced further through the addition of lime and consequently to improve the potential for chemical precipitation of the selected metals. The in situ removal experiment was accomplished through a set of seven columns filled with the sampled soil with varying contents of lime (i.e., from 0% to 6% by dry weight). Fresh leachate of low pH was added to the columns on a daily basis. Concentrations of manganese and zinc were measured in the influent and effluent during 40 days when biological clogging resulted in a condition of almost no outflow in the columns. The results indicated a substantial increase in removal efficiency through the addition of lime to the daily cover soil. Desorption resulting from the low pH of fresh leachate occurs at later stages compared to the column with no lime addition. PMID- 17125986 TI - Evaluation of investments in recycling centres for construction and demolition wastes in Brazilian municipalities. AB - There are very few construction and demolition (C&D) waste recycling centres in Brazil. To encourage the building and operation of new units, data were collected and analysed relating to C&D waste management and recycling in Brazil. Based on the results of this analysis, a conceptual model is presented for conducting viability studies of future C&D waste recycling centres. Applying this model to verify the viability of private recycling centres, the results show that under current market conditions in Brazil, C&D waste recycling centres are not financially feasible based solely on revenue from the sale of processed products. Nevertheless, under the same market conditions, the recycling centres could be economically viable for public authorities depending on the particular circumstances of each municipality. The feasibility, however, depends on continuity and the production volume reached. The conceptual model, the results of its applications and the discussions about the experiences of existing centres can strongly support public authorities and private initiatives in their decision making about investments in Brazil and in other developing countries. PMID- 17125987 TI - Dioxin emissions from thermal waste management in Medellin, Colombia: present regulation status and preliminary results. AB - Preliminary results of a study undertaken to characterize dioxin and furan releases from waste incineration plants operating in Medellin-Colombia are presented. Emission and fly ash samples were collected from representative plants burning medical and industrial residues to characterize PCDD/PCDF levels. Analyses were carried out following European standards for stationary gas emissions EN-1948:1996 and US EPA 1613 (fly ashes). Final extracts were analyzed by both high resolution gas chromatography coupled to high resolution mass spectrometry (HRGC-HRMS) and high resolution gas chromatography coupled to ion trap tandem mass spectrometry (HRGC-ITMS/MS). Preliminary results revealed emission levels of 1-30.3 ng I-TEQ/Nm3 depending on whether or not an air pollution control system (APCS) was installed. Fly ashes contained 8.5-68 ng I TEQ/g. Critical issues that should be addressed in an assessment and in developing waste management plans in Colombia are discussed. PMID- 17125988 TI - Interferometric biosensor based on planar optical waveguide sensor chips for label-free detection of surface bound bioreactions. AB - A label free optical biosensor based on a free-space Young interferometer configuration is presented. Commercial planar Ta(2) O(5) waveguides are used as sensing elements and allow the investigation of surface bound bioreactions like immunoreactions or biological affinity systems. Design criteria are discussed and a detailed characterization of the sensor performance is presented. The developed interferometer yields an effective refractive index resolution of 9 x 10(-9), corresponding to a surface coverage of approximately 13 fg/mm(2). The performance of the system is characterized by two different affinity systems: the antibody antigen complex protein G-immunoglobulin G is used as a model system for monitoring reaction kinetics. Further measurements on a silanized surface show the formation of a streptavidin monolayer on a biotinylated surface. PMID- 17125989 TI - Recombinant factor VIIa in massive postpartum haemorrhage. AB - Massive postpartum haemorrhage is a major cause of maternal and fetal morbidity and mortality. Management mainstays include transfusion therapy, uterotonic agents and surgery. The "off label" use of recombinant activated factor VII appears to have an evolving role in the management of massive postpartum haemorrhage refractory to conventional treatments. The current literature is reviewed. PMID- 17125990 TI - "The future of the midwife depends on her power to relieve pain". The rise and fall of the Analgesia in Childbirth Bill (1949). AB - The Analgesia in Childbirth Bill (1949) remains the only attempt in British history to give mothers the legal right to pain relief in childbirth. It provides a useful introduction to the major themes in the history of British obstetric anaesthesia and analgesia under the National Health Service. This paper places the bill in the dual contexts of long-standing concern over the status of British midwives and persistent governmental squabbling over the cost of the National Health Service. For its sponsors, the National Birthday Trust Fund, the bill provided an opportunity both to improve the availability of analgesia to mothers in childbirth and to reassert their dominance as leaders in the field of analgesia research. For the Ministry of Health under a Labour Minister, Aneurin Bevan, however, the bill represented both a (concealed) Conservative attack on the newly socialized British medical system and a further burden on the already overtaxed coffers of the National Health Service. Bevan's opposition to, and eventual defeat of, this enormously popular bill found its roots in the bill's economic implications rather than its clinical or humanitarian consequences. PMID- 17125991 TI - Effect of nitrous oxide on propofol requirement during target-controlled infusion for oocyte retrieval. AB - BACKGROUND: Oocyte (egg) retrieval for in-vitro fertilization is a relatively short procedure, usually performed as an outpatient. Propofol is a suitable anesthetic agent. Target-controlled infusion is a recently developed system that aids rapid recovery from propofol anesthesia. This study sought to determine the target concentration of propofol required to prevent movement in 50% (Cp50) and 95% (Cp95) of women during oocyte retrieval, and investigated whether supplemental nitrous oxide (N2O) modified the Cp50 and Cp95. METHODS: Forty-seven women scheduled for oocyte retrieval were randomly selected to receive either O2 air mixture (control group; n = 23) or 50% O2-N2O mixture (Nitrous oxide group; n = 24). Propofol was infused using a target-controlled infusion system with the concentration determined by up-down sequential allocation using 0.5 microg/mL step size. Transvaginal oocyte retrieval was performed after reaching target blood concentration. Patient responses to oocyte retrieval were noted as either no movement or movement. RESULTS: Using target-controlled infusion the Cp50 was 4.1 microg/mL in the control group and 3.3 microg/mL in the nitrous oxide group. Calculated Cp95 values were 4.0 microg/mL and 5.1 microg/mL with and without 50% nitrous oxide respectively. CONCLUSIONS: The Cp50 value for target-controlled infusion propofol during oocyte retrieval was significantly reduced by a factor of 1.24 (95% CI 1.07-1.44) with the use of 50% nitrous oxide. PMID- 17125992 TI - Cardiomyopathy in pregnancy and caesarean section: four case reports. AB - We present the clinical details of four women with cardiomyopathy who required caesarean section. Two women had peripartum cardiomyopathy and two had hypertrophic obstructive cardiomyopathy, one of whom has had two caesarean sections. Those with peripartum cardiomyopathy were more compromised than those with hypertrophic obstructive cardiomyopathy. Co-operation between obstetric and cardiac anaesthetists ensured optimum experience was available. An incremental combined spinal-epidural technique with invasive monitoring was used for three women and one received general anaesthesia. The risks and benefits of different anaesthetic techniques are discussed. PMID- 17125993 TI - Broken needle complicating spinal anaesthesia. PMID- 17125994 TI - Combined spinal-epidural anesthesia with very low dose hyperbaric levobupivacaine for cesarean section in a preeclamptic patient. PMID- 17125995 TI - A randomized trial of crystalloid versus colloid solution for prevention of hypotension during spinal or low-dose combined spinal-epidural anesthesia for elective cesarean delivery. AB - BACKGROUND: Spinal anesthesia for cesarean delivery is commonly associated with hypotension and nausea and vomiting, and preload with crystalloid or colloid solution is widely recommended. Low-dose spinal via the combined spinal-epidural technique appears to cause less hypotension and nausea and vomiting. The aim of this study was to investigate whether the combined use of colloid preload and combined spinal-epidural technique might further reduce the rates of these symptoms. METHODS: Women undergoing elective cesarean delivery were randomly allocated to one of four groups (50 in each) to receive crystalloid preload before spinal anesthesia, colloid preload before spinal anesthesia, crystalloid preload before combined spinal-epidural anesthesia, and colloid preload before combined spinal-epidural anesthesia. The incidences of hypotension and nausea and vomiting were compared. Spinal anesthesia was performed with 0.5% hyperbaric bupivacaine 9 mg and fentanyl 20 microg, and combined spinal-epidural anesthesia with 0.5% hyperbaric bupivacaine 6 mg + fentanyl 20 microg followed by epidural injection of 0.25% bupivacaine 10 mL. RESULTS: The frequencies of hypotension were 44%, 18%, 24%, and 20% in crystalloid preload-spinal anesthesia, colloid preload-spinal anesthesia, crystalloid preload-combined spinal epidural anesthesia, and colloid preload-combined spinal epidural anesthesia groups, respectively. The frequencies of nausea and vomiting were 20%, 2%, 8%, and 4% in respective groups. CONCLUSION: Colloid preload and low-dose spinal anesthesia alone or in combination lowered the incidences of hypotension and nausea. However, the combination of two methods failed to demonstrate further decreases in the incidence of the symptoms compared to the colloid-spinal anesthesia or crystalloid-combined spinal-epidural anesthesia groups. PMID- 17125996 TI - Anaesthesia for caesarean section in a woman with lung cancer: case report and review. AB - A 33-year-old woman pregnant with twins was diagnosed with metastatic lung cancer during pregnancy. Her multidisciplinary care raised many medical and ethical issues. To help decide on the best anaesthetic technique for caesarean section, a literature search of published case reports of pregnancy associated with lung cancer was performed. Thirty-five cases of primary lung cancer associated with pregnancy were found. Anaesthetic technique was reported in only five of the 20 patients who underwent caesarean section: one spinal, three epidurals and one general anaesthetic. Of the 11 patients who delivered vaginally, only one was reported to have received epidural analgesia. As published data regarding anaesthesia and analgesia are limited for women with lung cancer in pregnancy, we describe our perioperative approach and review the potential challenging aspects of management in a pregnant patient with metastatic lung cancer. PMID- 17125997 TI - Dose response to intrathecal diamorphine for elective caesarean section and compliance with a national audit standard. AB - BACKGROUND: This double-blind randomised controlled trial investigated the most appropriate dose of intrathecal diamorphine to use with high-dose diclofenac as part of a multimodal analgesic regimen for caesarean section under subarachnoid block. We also wished to establish whether it was possible to satisfy the Royal College of Anaesthetists postoperative pain audit recommendation for this patient group. METHODS: One hundred and twenty patients presenting for elective caesarean section under subarachnoid block were recruited and divided into four groups. Treatment was standard except that patients were given either placebo or one of three different doses of intrathecal diamorphine (100 microg, 200 microg or 300 microg). All patients were given regular paracetamol, high-dose diclofenac and an hourly subcutaneous diamorphine regimen for breakthrough pain. RESULTS: There was a dose-dependent improvement in analgesia with intrathecal diamorphine. Only 37.9% of patients given 300 microg of intrathecal diamorphine had a visual analogue pain score of 3/10 or less throughout the study. There was a dose dependent increase in the incidence of itching with intrathecal diamorphine although the incidence of nausea and vomiting was similar between groups. CONCLUSIONS: We found that for elective caesarean section under subarachnoid block with high dose diclofenac, analgesia was optimal with 300 microg of intrathecal diamorphine. Even the highest dose of intrathecal diamorphine did not achieve the Royal College of Anaesthetists postoperative audit target that 90% of patients should have a pain score of no more than 3/10. We believe that this target is too arduous. PMID- 17125998 TI - Effect of sufentanil on minimum local analgesic concentrations of epidural bupivacaine, ropivacaine and levobupivacaine in nullipara in early labour. AB - BACKGROUND: The aim was to assess the effect of epidural sufentanil on relative analgesic potencies of epidural bupivacaine, ropivacaine and levobupivacaine by determining the minimum local analgesic concentrations during labour. METHODS: In a randomised, double-blind study, 171 parturients were allocated to one of six groups receiving a 10-mL bolus of bupivacaine, ropivacaine or levobupivacaine alone or with sufentanil 0.75 microg/mL. The concentration of local anaesthetic was determined by the response of the previous parturient using up-down sequential allocation starting at a concentration of 0.13% wt/vol with a testing interval of 0.01%. Effective analgesia was defined as a visual analogue pain score < or = 15/100 mm within 30 min and lasting for 30 min. Median effective concentrations were estimated and two-sided P < 0.05 was significant. RESULTS: Local anaesthetic concentration, use of sufentanil and local anaesthetic drug were independent significant predictors of effective and ineffective analgesia. Bupivacaine was significantly more potent than levobupivacaine and ropivacaine. The relative potency ratios without sufentanil of 0.77:0.83:1.00 were reduced to 0.36:0.38:1.00 by the addition of sufentanil. The major factor influencing local anaesthetic requirements was the addition of sufentanil, which reduced overall requirements by a factor of 4.2 (95% CI 3.6-4.8); this effect was proportionately more enhanced for bupivacaine. CONCLUSIONS: Local anaesthetic requirements for bupivacaine, levobupivacaine and ropivacaine follow an analgesic potency hierarchy. Any potency differences are small when compared to the effect of sufentanil, which resulted in a four-fold reduction in local anaesthetic requirements. Sufentanil may also enhance the potency differences between bupivacaine and the two S-enantiomer agents. PMID- 17125999 TI - Evidence-based case report for analgesic and anesthetic management of a parturient with Ebstein's Anomaly and Wolff-Parkinson-White syndrome. AB - The rare congenital heart defect of Ebstein's anomaly is characterized by downward displacement and elongation of the septal cusp of tricuspid valve. As a result, it is often associated with a thin-walled and poorly contractile right ventricle, an enlarged atrium, tricuspid regurgitation, atrial septal defect with intracardiac shunt, pulmonary hypertension and tachyarrhythmia. Published reports or studies on analgesic and/or anesthetic management of laboring parturients with Ebstein's anomaly are limited. We present an evidence-based case report of the successful management of an obese laboring parturient with Ebstein's anomaly and Wolff-Parkinson-White syndrome under epidural analgesia. We reviewed the literature and present the best evidence available or its lack of, and the reasoning in supporting the choice of the analgesic and anesthetic management. PMID- 17126000 TI - Successful spinal blockade in a parturient with myotonia congenita. PMID- 17126001 TI - Anaesthetic management for caesarean section in moyamoya disease. PMID- 17126002 TI - Failed intubation in obstetrics: a self-fulfilling prophecy? PMID- 17126003 TI - An unusual intracranial tumour presenting in pregnancy. AB - We describe a patient who presented in late pregnancy with deteriorating neurological status due to an intracranial capillary haemangioma causing mass effect and raised intracranial pressure. She became confused and uncooperative leading to practical difficulties in performing adequate radiological imaging. Decision regarding timing of delivery and craniotomy was not straightforward and required discussion between the neurosurgeon, obstetrician and anaesthetist based on assessment of fetal maturity and the need to perform a craniotomy to excise what was initially thought to be a meningioma. Caesarean section was performed under general anaesthesia. The tumour was resected three weeks later. Management of obstetric patients with brain tumours is complex, requiring knowledge of the physiological effects of pregnancy on tumour size and labour on intracranial pressure. Both of these may influence the choice of labour analgesia or anaesthesia for caesarean section. Anaesthetists must be aware of the difficulties of radiological imaging during pregnancy, particularly in confused patients. The conflicting requirements of general anaesthesia for craniotomy and caesarean section should be considered. PMID- 17126005 TI - Post dural puncture headache in obstetric patients: experience from a West African teaching hospital. AB - BACKGROUND: This prospective, non-randomised study examined the frequency and severity of post dural puncture headache in 96 Ghanaian women who consented to spinal anaesthesia for caesarean section at the Korle Bu Teaching Hospital, Accra, Ghana. METHOD: Spinal anaesthesia was performed using 22-gauge (n = 22), 25-gauge (n = 46) or 26-gauge (n = 38) Quincke needles. Patients were followed up to determine the incidence and severity of post spinal headache. RESULT: The overall incidence of post dural puncture headache was 8.3%, but was significantly higher (33%) in patients in whom 22-gauge Quincke needles were used than in the other two groups (4% and 5% respectively: P = 0.003). Most patients rated their headache as mild to moderate on a 10-cm visual analogue scale. CONCLUSIONS: In view of the high incidence of headache and the need for treatment associated with the use of the 22-gauge Quincke needle, we recommend that this should not be used in the obstetric population. We are also aware that the incidence of post dural puncture headache could be further reduced by the use of small calibre pencil point needles but these are currently very expensive and many obstetric units in developing countries may not be able to afford them. PMID- 17126006 TI - Use of recombinant activated factor VII in massive obstetric haemorrhage. AB - Massive obstetric haemorrhage is a life-threatening emergency that remains a major cause of maternal mortality. Conventional management is aimed at optimising uterine tone, replacing circulating volume and blood products, and surgery to achieve haemostasis. Recently there have been numerous reports of the (unlicensed) use of recombinant activated factor VII in the management of major obstetric haemorrhage. We report our experience of using it in the treatment of major post-partum haemorrhage in four previously healthy parturients. The published reports of recombinant activated factor VII use in post-partum haemorrhage (unrelated to pre-existing coagulopathies) are compared. PMID- 17126008 TI - Anesthesia for cesarean section in a patient with Holt-Oram syndrome. AB - Holt-Oram syndrome is a rare genetic disorder affecting the heart and upper limbs (atriodigital dysplasia). The manifestations of the limb defects may vary in severity from subtle carpal bone defects and triphalangeal thumb to digit aplasia and upper extremity phocomelia. Cardiac abnormalities include atrial and/or ventricular septal defects, anomalies in pulmonary venous return and various dysrhythmias. We present the anesthetic management of a parturient with this syndrome who underwent elective cesarean section and tubal ligation, conducted under combined spinal-epidural anesthesia with a low dose of intrathecal bupivacaine. Our goal was to avoid an excessively high sympathetic block or excessive sympathetic stimulation accompanied by potential deleterious effects on cardiac rhythm. Cardiac monitoring was continued in the postoperative period for 6 h because of the possibility of dysrhythmia. PMID- 17126009 TI - Combined low-dose spinal-epidural anaesthesia versus single-shot spinal anaesthesia for elective caesarean delivery. PMID- 17126010 TI - The anaesthetist sandwich. PMID- 17126011 TI - Failed intubation drill in obstetrics. PMID- 17126012 TI - Anaphylactic reactions during cesarean section. AB - Sensitization to latex seems to occur more frequently in women than in men. Obstetric and gynecological surgical procedures have recently been shown to be a common setting for latex anaphylaxis. We analyzed all the cases of anaphylactic reactions during 1240 cesarean sections in 2004; the patients were questioned for risk factors and underwent allergy testing for drugs and latex. Four patients had anaphylaxis under spinal anesthesia and in all cases it was due to latex allergy. Reported symptoms included facial edema, profuse sweating, itching, generalized erythematous rash and hypotension. Only one patient manifested a severe reaction which included bronchospasm, dyspnea, tachypnea and anaphylactic shock, requiring orotracheal intubation and epinephrine. Our data showed a high incidence (1:310) of intraoperative latex anaphylactic reactions in the one-year study period. This may be related to the very specific population (all women) in a very specific setting (obstetrics). To prevent anaphylactic reactions during obstetric surgery it is important to identify potential risk factors to include, for example atopy, adverse reactions to foods and latex items. If latex allergy is confirmed or strongly suspected, patients should be managed in a latex-safe environment. Premedication with antihistamines and steroids might be useful to further reduce the risk. After the delivery, specific desensitization may represent a good therapeutic option. PMID- 17126013 TI - Lignin from rice straw Kraft pulping: effects on soil aggregation and chemical properties. AB - Lignin contained in pulping liquor that is generated during the pulping process for papermaking is a disposal problem for the pulp and paper industry. Separating lignin and other organic components from pulping liquor with inorganic acids may improve its applicability to fields as a beneficial soil amendment while offering a potential disposal alternative. Sulfuric acid-precipitated lignin from rice straw pulping liquor applied at rates of 1.67 and 3.34 g C kg(-1) soil was incubated to evaluate its effects on soil properties over 8 weeks of incubation. Addition of this acid-precipitated lignin at these rates decreased soil pH by 0.24-0.53 units over 8 weeks of incubation, suggesting that this sulfuric acid precipitated lignin from pulping liquor may have potential as a soil acidifying agent. Soil electrical conductivity (EC) only increased by up to 0.36 d Sm(-1), but highest EC levels were less than 4 d Sm(-1), indicating that lignin applied at both rates would not cause salinity problems. Application of this lignin increased soil organic C by 1.46 and 3.13 g C kg(-1), and total soil N by 0.07 and 0.17 g N kg(-1) over the incubation period. Lignin improved the macroaggregation of >2mm size fraction, and increased wet microaggregate stability of >2mm and 0.5-0.25 mm aggregates compared to a nonamended control. The results of this study suggest that this acid-precipitated lignin from pulping liquor may have potential as a beneficial soil amendment. PMID- 17126014 TI - Reduction of Pb and Zn bioavailable forms in metal polluted soils due to paper mill sludge addition. Effects on Pb and Zn transferability to barley. AB - In the last few years solidification/stabilisation of acidic soils polluted by heavy metals with low-cost sorbents has been investigated. Paper mill sludges are produced in large amounts and their disposal is a serious environmental problem. The possibility was therefore studied of using paper mill sludge as a stabilizer to reduce the bioavailable metal forms in polluted soils and thus the transferability of metals to plants (barley). We first investigated the sorbing properties of paper mill sludge for Zn(II) and Pb(II) and then their fractionation both in a polluted soil and in the same soil amended with paper mill sludge in order to check the decrease in mobile forms. Finally in both soils we tested the uptake of two metals by common barley in order to assess the performance of soil remediation from an ecological point of view. The addition of paper mill sludge to a soil contaminated by lead and zinc induces a decrease in the mobile forms of both metals, probably due to the presence in sludge of organic matter and kaolinite, which are able to bind the metals very strongly. The decrease in the mobile forms, which are the most readily available for uptake by plants, corresponds to a decrease in plant uptake. PMID- 17126015 TI - 5-Heteroatom substituted pyrazoles as canine COX-2 inhibitors. Part III: molecular modeling studies on binding contribution of 1-(5-methylsulfonyl)pyrid-2 yl and 4-nitrile. AB - The structure-activity relationship toward canine COX-1 and COX-2 in vitro whole blood activity of 4-hydrogen versus 4-cyano substituted 5-aryl or 5-heteroatom substituted N-phenyl versus N-2-pyridyl sulfone pyrazoles is discussed. The differences between the pairs of compounds with the 4-nitrile pyrazole derivatives having substantially improved in vitro activity are highlighted for both COX-2 and COX-1. This difference in activity may be due to the contribution of the hydrogen bond of the 4-cyano group with Ser 530 as shown by our molecular modeling studies. In addition, our model suggests a potential contribution from hydrogen bonding of the pyridyl nitrogen to Tyr 355 for the increased activity over the phenyl sulfone analogs. PMID- 17126016 TI - Design and synthesis of conformationally constrained tri-substituted ureas as potent antagonists of the human glucagon receptor. AB - A series of conformationally constrained tri-substituted ureas were synthesized, and their potential as glucagon receptor antagonists was evaluated. This effort resulted in the identification of compound 4a, which had a binding IC50 of 4.0 nM and was shown to reduce blood glucose levels at 3 mg/kg in glucagon-challenged mice containing a humanized glucagon receptor. Compound 4a was efficacious in correcting hyperglycemia induced by a high fat diet in transgenic mice at an oral dose as low as 3 mg/kg. PMID- 17126017 TI - Control of vertical components of gait during initiation of walking in normal adults and patients with progressive supranuclear palsy. AB - Gait and balance disorders are common in patients with parkinsonian syndromes, but the pathophysiology of these symptoms is still poorly understood. This study examined the initiation of gait in patients with progressive supranuclear palsy (PSP, n=10), characterized by the presence of severe postural instability, and controls (n=43). We used a force plate to measure the step length, and the antero posterior and vertical velocities of the centre of gravity (CG) during gait initiation, in natural and fast gait conditions. In controls, during the swing limb period, there was a fall in the CG, which was reversed before foot contact. When controls were asked to walk faster, the fall in the CG increased but the ability to brake the fall in CG was unchanged. In PSP patients, length and maximal velocity of the first step were reduced compared to controls and no anticipatory braking in the CG fall occurred prior to the foot-contact, especially in the fast gait condition. The results suggest that normal subjects actively brake the fall in the CG prior to foot-contact. How this phenomenon participates in balance control is unknown. We hypothesize that the absence of active braking of the fall in the CG prior to foot-contact, observed in PSP patients, could contribute to the postural instability, characteristic of this disorder. PMID- 17126018 TI - Exploring the active site of phenylethanolamine N-methyltransferase with 1,2,3,4 tetrahydrobenz[h]isoquinoline inhibitors. AB - 1,2,3,4-Tetrahydrobenz[h]isoquinoline (THBQ, 11) is a potent, inhibitor of phenylethanolamine N-methyltransferase (PNMT). Docking studies indicated that the enhanced PNMT inhibitory potency of 11 (hPNMT K(i)=0.49microM) versus 1,2,3,4 tetrahydroisoquinoline (5, hPNMT K(i)=5.8microM) was likely due to hydrophobic interactions with Val53, Met258, Val272, and Val269 in the PNMT active site. These studies also suggested that the addition of substituents to the 7-position of 11 that are capable of forming hydrogen bonds to the enzyme could lead to compounds (14-18) having enhanced PNMT inhibitory potency. However, these compounds are in fact less potent at PNMT than 11. Furthermore, 7-bromo-THBQ (19, hPNMT K(i)=0.22mM), which has a lipophilic 7-substituent that cannot hydrogen bond to the enzyme, is twice as potent at PNMT than 11. This once again illustrates the limitations of docking studies for lead optimization. PMID- 17126019 TI - Design and study of some novel ibuprofen derivatives with potential nootropic and neuroprotective properties. AB - Six novel ibuprofen derivatives and related structures, incorporating a proline moiety and designed for neurodegenerative disorders, are studied. They possess anti-inflammatory properties and three of them inhibited lipoxygenase. One compound was found to inhibit cyclooxygenase (COX)-2 production in spleenocytes from arthritic rats. The HS-containing compounds are potent antioxidants and one of them protected against glutathione loss after cerebral ischemia/reperfusion. They demonstrated lipid-lowering ability and seem to acquire low gastrointestinal toxicity. Acetylcholinesterase inhibitory activity, found in two of these compounds, may be an asset to their actions. PMID- 17126020 TI - Study on dual-site inhibitors of acetylcholinesterase: Highly potent derivatives of bis- and bifunctional huperzine B. AB - Natural (-)-huperzine B (HupB), isolated from Chinese medicinal herb, displayed moderate inhibitory activity of acetylcholinesterase (AChE). Based on the active dual-site of AChE, a series of novel derivatives of bis- and bifunctional HupB were designed and synthesized. The AChE inhibition potency of most derivatives of HupB was enhanced about 2-3 orders of magnitude as compared with the parental HupB. Among bis-HupB derivatives, 12h exhibited the most potent in the AChE inhibition and has been evaluated for its pharmacological actions in vivo on ChE inhibition, cognitive enhancement, and neuroprotection. The docking study on the bis-HupB derivatives 12 series with TcAChE has demonstrated that the ligands bound to the dual-site of the enzyme in different level. PMID- 17126021 TI - Synthesis of novel quinolone and quinoline-2-carboxylic acid (4-morpholin-4-yl phenyl)amides: a late-stage diversification approach to potent 5HT1B antagonists. AB - Multiparallel amenable syntheses of 6-methoxy-8-amino-4-oxo-1,4-dihydroquinoline 2-carboxylic acid-(4-morpholin-4-yl-phenyl)amides (I) and 4-amino-6-methoxy-8-(4 methyl-piperazin-1-yl)-quinoline-2-carboxylic acid (4-morpholin-4-yl phenyl)amides (II) which facilitate late-stage diversification at the 8-position of (I) and at the 4- and 8-positions of (II) are described. The resulting novel series were determined to contain potent 5HT(1B) antagonists. Preliminary SAR data are presented. PMID- 17126022 TI - Tetrahydrobenzotriazines as a new class of nematocide. AB - Tetrahydrobenzotriazines, a novel class of heterocyclic compounds, were synthesized. Examination of their biological activities resulted in the discovery that some of them possess potent nematocidal activity. PMID- 17126023 TI - Automated measurement of 224Ra and 226Ra in water. AB - We present a new simple approach for automated, non-destructive measurement of the alpha-emitting radium isotopes ((223)Ra, (224)Ra, and (226)Ra) in water based on the emanation of their respective radon daughters ((219)Rn, (220)Rn, and (222)Rn). The method combines the high adsorption uptake of MnO(2) Resin for radium (K(d)=2.4 x 10(4)ml/g) over a wide pH range with the simplicity of the activity registration using a commercial radon-in-air analyzer (RAD7, DURRIDGE Company, Inc). Radium is first adsorbed onto the MnO(2) Resin by passing a water sample through the resin packed in a gas-tight glass cartridge. The same cartridge is then connected to the radon analyzer via a simple tubing system to circulate air through the resin and a drying system. The efficiency of the proposed system is determined by running standards prepared in the same manner. Our results indicate that the efficiency for (226)Ra is >22% if both (218)Po and (214)Po counts are collected. This is comparable with typical efficiencies for alpha spectrometry but with much less sample preparation. We estimate that an MDA of 0.8 pCi/L for (226)Ra may be obtained with this new approach using a 1L water sample and less than 4h of counting. PMID- 17126024 TI - Pulmonary resection in octogenarians. PMID- 17126025 TI - Evaluation of low energy CID and ECD fragmentation behavior of mono-oxidized thio ether bonds in peptides. AB - Thio-ether bonds in the cysteinyl side chain of peptides, formed with the most commonly used cysteine blocking reagent iodoacetamide, after conversion to sulfoxide, releases a neutral fragment mass in a low-energy MS/MS experiment in the gas phase of the mass spectrometer [6]. In this study, we show that the neutral loss fragments produced from the mono-oxidized thio-ether bonds (sulfoxide) in peptides, formed by alkyl halide or double-bond containing cysteine blocking reagents are different under low-energy MS/MS conditions. We have evaluated the low-energy fragmentation patterns of mono-oxidized modified peptides with different cysteine blocking reagents, such as iodoacetamide, 3 maleimidopropionic acid, and 4-vinylpyridine using FTICR-MS. We propose that the mechanisms of gas-phase fragmentation of mono-oxidized thio-ether bonds in the side chain of peptides, formed by iodoacetamide and double-bond containing cysteine blocking reagents, maleimide and vinylpyridine, are different because of the availability of acidic beta-hydrogens in these compounds. Moreover, we investigated the fragmentation characteristics of mono-oxidized thio-ether bonds within the peptide sequence to develop novel mass-spectrometry identifiable chemical cross-linkers. This methionine type of oxidized thio-ether bond within the peptide sequence did not show anticipated low-energy fragmentation. Electron capture dissociation (ECD) of the side chain thio-ether bond containing oxidized peptides was also studied. ECD spectra of the oxidized peptides showed a greater extent of peptide backbone cleavage, compared with CID spectra. This fragmentation information is critical to researchers for accurate data analysis of this undesired modification in proteomics research, as well as other methods that may utilize sulfoxide derivatives. PMID- 17126026 TI - Genomic mechanisms and measurement of structural and numerical instability in cancer cells. AB - The progression to cancer is often associated with instability and the acquisition of genomic heterogeneity, generating both clonal and non-clonal populations. Chromosomal instability (CIN) describes the excessive rate of numerical and structural genomic change in tumors. Mitotic segregation errors strongly influences copy number, while structural aberrations can occur at unstable genomic regions, or through aberrant DNA repair or methylation. Combined molecular cytogenetic analyses can evaluate cell-to-cell variation, and define the complexity of numerical and structural alterations. Because structural change may occur independently of numerical alteration, we propose the term structural chromosomal instability [(S)-CIN] to distinguish numerical from structural CIN. PMID- 17126027 TI - Reduced reactivity to novelty, impaired social behavior, and enhanced basal synaptic excitatory activity in perforant path projections to the dentate gyrus in young adult mice deficient in the neural cell adhesion molecule CHL1. AB - The neural cell adhesion molecule CHL1 is implicated in neural development in the mouse and has been related to psychiatric disorders in humans. Here we report that mice constitutively deficient for CHL1 display reduced reactivity to environmental stimuli and reduced expression of social behaviors, whereas cognitive, motor and olfactory functions are normal. Basal synaptic transmission and plasticity in seven major excitatory connections in the hippocampus were analyzed to test whether dysfunctions in this brain region, which controls complex behaviors, correlate with the behavioral alterations of CHL1 deficient mice. We found that basal synaptic transmission in lateral and medial perforant path projections to the dentate gyrus is elevated in CHL1-deficient mice. Taking in consideration the function of these synapses in processing information from cortical areas, we hypothesize that constitutive ablation of CHL1 leads to reduced capability to react to external stimuli due to dysfunctions in the dentate gyrus. PMID- 17126028 TI - Interactive effects of C3, cyclic AMP and ciliary neurotrophic factor on adult retinal ganglion cell survival and axonal regeneration. AB - We tested whether combined therapy involving Rho inactivation, elevation of cAMP and supply of ciliary neurotrophic factor (CNTF) (i) increased axotomized adult retinal ganglion cell (RGC) survival and (ii) promoted axonal regeneration into peripheral nerve (PN) autografted onto the cut optic nerve. PN-grafted eyes were injected with combinations of a Rho-inactivating enzyme C3 transferase (C3-11), CNTF and a cell-permeant analogue of cAMP (CPT-cAMP). Four weeks after PN transplantation, RGC survival was quantified using beta-III tubulin immunohistochemistry. Regeneration was assessed using retrograde fluorogold tracing and pan-neurofilament immunostaining of grafts. Treatment with C3-11 increased RGC survival but co-injection with CPT-cAMP, CNTF or combined CNTF/CPT cAMP did not further enhance RGC viability. There were greater numbers of regenerating RGCs after multiple C3-11 injections and regeneration was further and significantly increased after intravitreal injections of all three factors. In the combined C3-11/CNTF/CPT-cAMP treatment group about 15% of RGCs remained viable of which more than half regenerated an axon. These data emphasize the power of combinatorial pharmacotherapeutic and transplant strategies in the treatment of neurotrauma. PMID- 17126029 TI - Effect of osmotic stress and heat shock in recombinant protein overexpression and crystallization. AB - Overexpressed recombinant proteins in bacteria often tend to misfold and accumulate as soluble aggregates and/or inclusion bodies. A strategy for improving the level of expression of recombinant proteins in a soluble native form is to increase the cellular concentration of osmolytes or of chaperones. This can be accomplished by growing the bacterial cells in the presence of high salt, sorbitol, and betaine as well as exposing the cells to a heat shock step. Our results suggest that by growing the cells under varied conditions one may be able to express targets as soluble proteins (from previously insoluble targets) and to improve the chances of their crystallization. PMID- 17126030 TI - Improved performance of column chromatography by arginine: dye-affinity chromatography. AB - Arginine has been effectively used in various column chromatographies for improving recovery and resolution, and suppressing aggregation. Here, we have tested the effectiveness of arginine as an eluent in dye-affinity column chromatography using Blue-Sepharose, which binds enzymes requiring adenyl containing cofactors (e.g., NAD). A common eluent, NaCl, showed a broad elution peak with low recovery of lactate dehydrogenase, at most approximately 60% using 2M salt. The recovery decreased as the NaCl concentration was either decreased or increased; i.e., the recovery was maximum at 2M. On the contrary, addition of arginine to the eluent resulted in more than 80% recovery above 0.5M and the recovery was nearly independent of the arginine concentration. The elution peak was much sharper with arginine, leading to elution of more concentrated protein solution. Successful elution of proteins bound to the ATP-agarose resins by arginine was also described. PMID- 17126031 TI - Isolation and crystallization of a unique size category of recombinant Rabies virus Nucleoprotein-RNA rings. AB - In order to study the packaging of rabies virus RNA inside the viral nucleocapsid, rabies nucleoprotein was expressed in insect cells. In the cells, it binds to cellular RNA to form long, helical or short circular complexes, depending on the length of the bound RNA. The circular complexes contained from 9 up to 13 N-protomers per ring. Separation of the rings into defined size classes was impossible through regular column chromatographies or gradient centrifugation. The size classes could be separated by native polyacrylamide gel electrophoresis. A large-scale separation was achieved with a 4% native gel using a preparative electrophoresis apparatus. Crystallization trials were set up with N-RNA rings from three size classes and crystals were obtained in all cases. The best diffracting crystals, diffracting up to 6A, contained rings with 11 N protomers plus an RNA molecule of 99 nucleotides. The diffraction limit was improved to 3.5A by air dehydration prior to flash freezing. PMID- 17126032 TI - Molecular basis of organization of collagen fibrils. AB - The collagen fibrils are formed by self-assembly of individual collagen molecules, but the mechanism that drives their orderly packing during fibril formation is not clearly defined. To identify structural determinants critical for the D-periodic alignment of collagen molecules we employed three sets of genetically engineered collagen II variants: (i) a set in which domains corresponding to the specific D periods have been purposely deleted, (ii) a set of collagen variants consisting of tandem repeats of a specific D period, and (iii) a set lacking definite fragments of the D4 period. All collagen variants were analyzed for their ability to assemble into D-periodic fibrils. Even though all genetically engineered collagen variants differ significantly from the wild type collagen II, most of them were able to form filamentous structures. The D periodic banding pattern, an indication of the staggered arrangement of collagen monomers, however, occurred only when the D1, D4, and D0.4 domains of interacting collagen monomers could potentially cluster together to form a triad through telopeptide-mediated binding. Our results identify a critical step in the formation of collagenous matrices and provide experimental evidence for the active involvement of the N-terminal and C-terminal regions of fibrillar collagens in this process. PMID- 17126033 TI - Mechanical and morphological properties of the triceps surae muscle-tendon unit in old and young adults and their interaction with a submaximal fatiguing contraction. AB - The purposes of this study were to examine (a) whether the morphological properties of the muscle gastrocnemius medialis (GM) contribute to the known enhanced muscle fatigue resistance during submaximal sustained isometric plantar flexion contraction of old compared to young adults and (b) whether a submaximal fatiguing contraction differently affects the mechanical properties of the GM tendon and aponeurosis of old and young adults. Fourteen old and 12 young male subjects performed maximal voluntary isometric plantar flexions (MVC) on a dynamometer before and after a submaximal fatiguing task (40% MVC). Moments and EMG signals from the gastrocnemius medialis and lateralis, soleus and tibialis anterior muscles were measured. The elongation of the GM tendon and aponeurosis and the morphological properties of its contractile element were examined by means of ultrasonography. The old adults showed lower maximal ankle joint moment, stiffness and fascicle length in both tested conditions. The submaximal fatiguing contraction did not affect the force-strain relationship of the GM tendon and aponeurosis of either young or old adults. The time to task failure was longer for the old adults and was strongly correlated with the fascicle length (r(2)=0.50, P<0.001). This provides evidence on that the lower ratio of the active muscle volume to muscle force for the old adults might be an additional mechanism contributing to the known age related increase in muscle fatigue resistance. PMID- 17126034 TI - Varieties of grapheme-colour synaesthesia: a new theory of phenomenological and behavioural differences. AB - Recent research has suggested that not all grapheme-colour synaesthetes are alike. One suggestion is that they can be divided, phenomenologically, in terms of whether the colours are experienced in external or internal space (projector associator distinction). Another suggestion is that they can be divided according to whether it is the perceptual or conceptual attributes of a stimulus that is critical (higher-lower distinction). This study compares the behavioural performance of 7 projector and 7 associator synaesthetes. We demonstrate that this distinction does not map on to behavioural traits expected from the higher lower distinction. We replicate previous research showing that projectors are faster at naming their synaesthetic colours than veridical colours, and that associators show the reverse profile. Synaesthetes who project colours into external space but not on to the surface of the grapheme behave like associators on this task. In a second task, graphemes presented briefly in the periphery are more likely to elicit reports of colour in projectors than associators, but the colours only tend to be accurate when the grapheme itself is also accurately identified. We propose an alternative model of individual differences in grapheme colour synaesthesia that emphasises the role of different spatial reference frames in synaesthetic perception. In doing so, we attempt to bring the synaesthesia literature closer to current models of non-synaesthetic perception, attention and binding. PMID- 17126035 TI - Unconscious processing embedded in conscious processing: evidence from gaze time on Chinese sentence reading. AB - The current study aims to separate conscious and unconscious behaviors by employing both online and offline measures while the participants were consciously performing a task. Using an eye-movement tracking paradigm, we observed participants' response patterns for distinguishing within-word-boundary and across-word-boundary reverse errors while reading Chinese sentences (also known as the "word inferiority effect"). The results showed that when the participants consciously detected errors, their gaze time for target words associated with across-word-boundary reverse errors was significantly longer than that for targets words associated with within-word-boundary reverse errors. Surprisingly, the same gaze time pattern was found even when the readers were not consciously aware of the reverse errors. The results were statistically robust, providing converging evidence for the feasibility of our experimental paradigm in decoupling offline behaviors and the online, automatic, and unconscious aspects of cognitive processing in reading. PMID- 17126036 TI - Quantitative basal CBF and CBF fMRI of rhesus monkeys using three-coil continuous arterial spin labeling. AB - A three-coil continuous arterial-spin-labeling technique with a separate neck labeling coil was implemented on a Siemens 3T Trio for quantitative cerebral blood flow (CBF) and CBF fMRI measurements in non-human primates (rhesus monkeys). The optimal labeling power was 2 W, labeling efficiency was 92+/-2%, and optimal post-labeling delay was 0.8 s. Gray matter (GM) and white matter (WM) were segmented based on T1 maps. Quantitative CBF were obtained in 3 min with 1.5 mm isotropic resolution. Whole-brain average DeltaS/S was 1.0-1.5%. GM CBF was 104+/-3 ml/100 g/min (n = 6, SD) and WM CBF was 45+/-6 ml/100 g/min in isoflurane anesthetized rhesus monkeys, with the CBF GM/WM ratio of 2.3+/-0.2. Combined CBF and BOLD (blood-oxygenation-level-dependent) fMRI associated with hypercapnia and hyperoxia were made with 8-s temporal resolution. CBF fMRI responses to 5% CO2 were 59+/-10% (GM) and 37+/-4% (WM); BOLD fMRI responses were 2.0+/-0.4% (GM) and 1.2+/-0.4% (WM). CBF fMRI responses to 100% O2 were -9.4+/-2% (GM) and -3.9+/ 2.6% (WM); BOLD responses were 2.4+/-0.7% (GM) and 0.8+/-0.2% (WM). The use of a separate neck coil for spin labeling significantly increased CBF signal-to-noise ratio and the use of small receive-only surface coil significantly increased signal-to-noise ratio and spatial resolution. This study sets the stage for quantitative perfusion imaging and CBF fMRI for neurological diseases in anesthetized and awake monkeys. PMID- 17126037 TI - Sequence of information processing for emotions based on the anatomic dialogue between prefrontal cortex and amygdala. AB - The prefrontal cortex and the amygdala have synergistic roles in regulating purposive behavior, effected through bidirectional pathways. Here we investigated the largely unknown extent and laminar relationship of prefrontal input-output zones linked with the amygdala using neural tracers injected in the amygdala in rhesus monkeys. Prefrontal areas varied vastly in their connections with the amygdala, with the densest connections found in posterior orbitofrontal and posterior medial cortices, and the sparsest in anterior lateral prefrontal areas, especially area 10. Prefrontal projection neurons directed to the amygdala originated in layer 5, but significant numbers were also found in layers 2 and 3 in posterior medial and orbitofrontal cortices. Amygdalar axonal terminations in prefrontal cortex were most frequently distributed in bilaminar bands in the superficial and deep layers, by columns spanning the entire cortical depth, and less frequently as small patches centered in the superficial or deep layers. Heavy terminations in layers 1-2 overlapped with calbindin-positive inhibitory neurons. A comparison of the relationship of input to output projections revealed that among the most heavily connected cortices, cingulate areas 25 and 24 issued comparatively more projections to the amygdala than they received, whereas caudal orbitofrontal areas were more receivers than senders. Further, there was a significant relationship between the proportion of 'feedforward' cortical projections from layers 2-3 to 'feedback' terminations innervating the superficial layers of prefrontal cortices. These findings indicate that the connections between prefrontal cortices and the amygdala follow similar patterns as corticocortical connections, and by analogy suggest pathways underlying the sequence of information processing for emotions. PMID- 17126038 TI - How long to scan? The relationship between fMRI temporal signal to noise ratio and necessary scan duration. AB - Recent advances in MRI receiver and coil technologies have significantly improved image signal-to-noise ratios (SNR) and thus temporal SNR (TSNR). These gains in SNR and TSNR have allowed the detection of fMRI signal changes at higher spatial resolution and therefore have increased the potential to localize small brain structures such as cortical layers and columns. The majority of current fMRI processing strategies employ multi-subject averaging and therefore require spatial smoothing and normalization, effectively negating these gains in spatial resolution higher than about 10 mm3. Reliable detection of activation in single subjects at high resolution is becoming a more common desire among fMRI researchers who are interested in comparing individuals rather than populations. Since TSNR decreases with voxel volume, detection of activation at higher resolutions requires longer scan durations. The relationship between TSNR, voxel volume and detectability is highly non-linear. In this study, the relationship between TSNR and the necessary fMRI scan duration required to obtain significant results at varying P values is determined both experimentally and theoretically. The results demonstrate that, with a TSNR of 50, detection of activation of above 2% requires at most 350 scan volumes (when steps are taken to remove the influence of physiological noise from the data). Importantly, these results also demonstrate that, for activation magnitude on the order of 1%, the scan duration required is more sensitive to the TSNR level than at 2%. This study showed that with voxel volumes of approximately 10 mm3 at 3 T, and a corresponding TSNR of approximately 50, the required number of time points that guarantees detection of signal changes of 1% is about 860, but if TSNR increases by only 20%, the time for detection decreases by more than 30%. More than just being an exercise in numbers, these results imply that imaging of columnar resolution (effect size=1% and assuming a TR of 1 s) at 3 T will require either 10 min for a TSNR of 60 or 40 min for a TSNR of 30. The implication is that at these resolutions, TSNR is likely to be critical for determining success or failure of an experiment. PMID- 17126039 TI - Brain dopamine transporter levels in treatment and drug naive adults with ADHD. AB - Attention deficit hyperactivity disorder (ADHD) is the most frequent psychiatric disorder in children, yet data are sparse on its pathophysiology. Particularly relevant are the dopamine transporters since these are the main targets of stimulant medications used for ADHD treatment. Though some imaging studies have shown increases in dopamine transporters in ADHD others have not and their role in the neurobiology of ADHD remains unclear. Here we investigate dopamine transporters in ADHD subjects with control of potentially confounding factors (previous medication and/or drug histories, comorbidity) and their association with clinical symptoms. Positron emission tomography and [11C]cocaine were used to measure dopamine transporters in 20 never medicated adults with ADHD and 25 controls. Dopamine transporters were lower in left caudate (13%, p < 0.05) and in left nucleus accumbens (p < 0.005) in ADHD subjects than in controls. In putamen dopamine transporters did not differ between groups but were associated with scores of inattention (Conners Adult Attention Rating Scale) both in ADHD subjects (p < 0.005) and in controls (p < 0.005). Thus, for a given transporter level the scores for inattention were on average five times greater in ADHD subjects than in controls. These results do not corroborate increases in dopamine transporters in ADHD subjects and show that in some they are reduced. It also provides evidence that dopamine transporter levels modulate attention but suggest that additional pathology (e.g., prefrontal or cingulostriatal pathways, noradrenergic neurotransmission) is necessary to account for the large differences in inattention observed between controls and ADHD subjects. PMID- 17126040 TI - Helicobacter pylori infection in epileptic patients. AB - It is well known that there might be an epidemiological association between Helicobacter pylori infection and extraintestinal diseases. This study aimed at determining H. pylori infection in epileptic patients. Forty-seven cryptogenic epileptic patients (Group 1) and 35 healthy people (Group 2) participated in this study. Presence of H. pylori infection was examined by H. pylori stool antigen (HpSA), H. pylori IgG, and IgM. HpSA was detected in 21 participants (44.6%) in Group 1 and in 3 participants (8.5%) in Group 2. H. pylori IgM was positive in 27 participants (57.4%) in Group 1 and in 8 participants (22.8%) in Group 2. H. pylori IgG was positive in 37 participants (78.7%) in Group 1 and in 13 participants (38%) in Group 2. The difference of rates of HpSA, H. pylori IgM and IgG in Groups 1 and 2 were found statistically significant (chi2=4.18, p=0.04; chi2=9.18, p=0.0017; chi2=14.58, p<0.001, respectively). We also compared presence of H. pylori infection between the epileptic patients with poor and good prognosis; HpSA positivity was detected in 15 (62.5%) of 24 and 6 (26%) of 23, respectively, and the differences were statistically significant (chi2=6.30, p=0.012). H. pylori IgM positivity was detected in 16 (66%) of 24 patients with poor prognosis and 11 (47.8%) of 23 patients with good prognosis (p>0.05). H. pylori IgG positivity was detected in 18 (75%) of 24 patients with poor prognosis and 19 (82.6%) of 23 patients with good prognosis. The differences of H. pylori IgM and IgG positivity rates in epileptic patients with poor and good prognosis were not found statistically significant (p>0.05). These results suggest a probable association between the acute H. pylori infection and epilepsy, especially with poor prognosis. PMID- 17126041 TI - Antimicrobial activity of different proteins and their fragments from Bacillus thuringiensis parasporal crystals against clostridia and archaea. AB - Proteins of parasporal crystals (Cry proteins) from entomopathogenic bacterium Bacillus thuringiensis (subspecies kurstaki, galleriae, tenebrionis) as well as some fragments of these proteins, obtained by limited proteolysis, are capable of antimicrobial action against anaerobic bacteria and archaea-Clostridium butyricum, Clostridium acetobutylicum and Methanosarcina barkeri. The MICs are 45 150 microg/mL. Electron microscopy showed that lysis of M. barkeri cells in the presence of 49kDa fragment of Cry3Aa toxin is generally similar to the bacterial cell lysis, which has been previously detected in the presence of Cry11A, Cry1Ab and other Cry proteins. The Cry1D-like toxin from crystals of B. thuringiensis subsp. galleriae has been put forward as an example of the supposition that cell wall and some of its components like teichoic acid and N-acetylgalactosamine have possible influence on Cry toxins, enhancing their antimicrobial activity. The possible ecological role of the antimicrobial activity of Cry proteins is also discussed. PMID- 17126042 TI - Differential physiological and developmental expression of the UapA and AzgA purine transporters in Aspergillus nidulans. AB - In this article we study the cellular expression of UapA and AzgA, the two major purine transporters of Aspergillus nidulans, by constructing strains expressing, from their native promoters, fully functional fluorescent (UapA-sGFP, AzgA-sGFP) or immunological (UapA-His) chimeric transporters. Epifluorescence microscopy and immunodetection showed that under different physiological conditions and during Aspergillus development: (i) UapA and AzgA expression in the plasma membrane becomes evident early during germination and remains at a significant basal level in mycelium, (ii) Neither of the two transporters is expressed in the stalk, the vesicle, the phialides and the conidiospores, but surprisingly, UapA is specifically and strongly expressed in the periphery of metulae, (iii) Both transporters are expressed in ascogenous hyphae and in hulle cells but not in cleistothecia or ascospores, (iv) Purine induction leads to approximately 4-fold increase in UapA and AzgA protein content in mycelium, compatible with an analogous increase at the transcriptional level, (v) Ammonium leads to removal of UapA, but not of AzgA, from the plasma membrane by sorting of the protein to the vacuole. PMID- 17126043 TI - Aortic stenosis is associated with elevated plasma levels of asymmetric dimethylarginine (ADMA). AB - OBJECTIVES: We tested the hypothesis that the presence of aortic stenosis (AS) is associated with elevation of plasma levels of asymmetric dimethylarginine (ADMA), a physiological inhibitor of nitric oxide synthase, a mediator and marker of endothelial dysfunction and an indicator of incremental cardiovascular risk. BACKGROUND: The presence of aortic sclerosis (ASC), the precursor of AS is independently associated both with endothelial dysfunction, and with incremental coronary event risk. It remains uncertain whether elevations of ADMA levels might mediate endothelial dysfunction in these conditions. METHODS: Forty two consecutive patients referred for echocardiography for evaluation of AS, who had calculated aortic valve areas of <1.4 cm(2) (AS group) were evaluated together with 42 consecutive age-matched referred patients (non-AS group). Plasma ADMA levels were measured by high-performance liquid chromatography (HPLC). Determinants of elevation of plasma ADMA levels were identified via stepwise multiple linear regression analysis. RESULTS: Plasma ADMA levels were not statistically different between the AS and non-AS group (median 0.59 vs 0.54 micromol/L, p=0.13, Mann-Whitney test) on univariate analysis. However, in backward stepwise multiple linear regression, the presence of AS was a significant predictor of elevated ADMA levels (p=0.04, 95% CI=0.001, 0.072). In addition, elevated plasma ADMA levels were also associated with history of atrial fibrillation (p=0.009, 95% CI=0.015, 0.100), and negatively associated with creatinine clearance (p=0.01, 95% CI=-0.002, 0.000), and the use of statin therapy (p=0.01, 95% CI=-0.081, -0.011). CONCLUSIONS: AS is independently associated with elevation of ADMA levels, beyond that implied by "conventional" risk factors for endothelial dysfunction. The clinical status of AS as an incremental marker of cardiovascular risk may reflect ADMA-mediated endothelial dysfunction. PMID- 17126044 TI - Nitric oxide inhibits depolarization-evoked glutamate release from rat cerebellar granule cells. AB - Nitric oxide (NO) modulates the release of various neurotransmitters, some of these are considered to be involved in neuronal plasticity that includes long term depression in the cerebellum. To date, there have been no reports on the modulation of the exocytotic release of neurotransmitters in the cerebellar granule cells (CGCs) by NO. The aim of this study was to investigate the effects of NO on the exocytotic release of glutamate from rat CGCs. Treatment with NO related reagents revealed that NO inhibited high-K(+)-evoked glutamate release. Clostridium botulinum type B neurotoxin (BoNT/B) attenuated the enhancement of glutamate release caused by NO synthase (NOS) inhibition; this indicates that NO acts on the high-K(+)-evoked exocytotic pathway. cGMP-related reagents did not affect the high-K(+)-evoked glutamate release. NO-related reagents did not affect Ca(2+) ionophore-induced glutamate release, suggesting that NO inhibits Ca(2+) entry through voltage-dependent Ca(2+) channels (VDCC). Monitoring of intracellular Ca(2+) revealed that NO inhibited high-K(+)-evoked Ca(2+) entry. L type VDCC blockers inhibited glutamate release and NO did not have an additive effect on the inhibition produced by the L-type VDCC blocker. The inhibition of the high-K(+)-evoked glutamate release by NO was abolished by a reducing reagent; this suggested that NO regulates the high-K(+)-evoked glutamate release from CGCs by redox modulation. PMID- 17126045 TI - Clinical characteristics and pain management among patients with painful peripheral neuropathic disorders in general practice settings. AB - Alleviating chronic pain is a global healthcare priority. Understanding the medical profile and current treatment patterns in patients with painful neuropathic disorders (PNDs) is crucial to the development of effective pain management strategies. Thus, our objective was to describe the demographic and clinical characteristics of persons with PNDs and their use of pain medications. Using the general practice research database, we categorized PNDs in two ways: Pure PNDs (which include diabetic neuropathy, postherpetic neuralgia, etc.; N=16,690) and Mixed PNDs (which include back/neck pain with neuropathic involvement; N=14,309). On average, PND patients were 55 years old (Pure, 55.4 [SD=16.9] years; Mixed, 54.3 [SD=16.4] years). Over a third had other chronic pain-related (Pure, 37.5%; Mixed, 37.1%) and nearly a quarter had non-pain related (Pure, 28.1%; Mixed, 24.1%) comorbidities. Use of medications with clinically demonstrated efficacy in PNDs was higher among patients with Pure PNDs (tricyclic antidepressants [Pure, 16.6%; Mixed, 10.1%]; 2nd generation antidepressants [Pure, 11.0%; Mixed, 9.7%]; and antiepileptics [Pure, 12.2%; Mixed, 2.6%]), whereas use of NSAIDs (Pure, 43.1%; Mixed, 65.2%) and opioids (Pure, 8.5%; Mixed, 14.3%) was higher among patients with Mixed PNDs. Average daily doses of select neuropathic pain-related medications among PND patients (Pure and Mixed) were lower than those recommended for neuropathic pain. Among both Pure and Mixed PND patients, use and doses of evidenced-based neuropathic pain-related medications was low, and lower than the use of NSAIDs (a medication class with no proven efficacy for PNDs) in each group, suggesting possible sub optimal neuropathic pain management among these patients. PMID- 17126046 TI - Adherence of Irish general practitioners to European guidelines for acute low back pain: a prospective pilot study. AB - There are no national low back pain (LBP) clinical guidelines in Ireland, and neither the level of adherence of General Practitioners (GPs) to the European guidelines, nor the cost of LBP to the patient and the state, have been investigated. A prospective pilot study was conducted on 54 consenting patients (18M, 36F: mean age (SD): 40.5 (14.3) years) with a new episode of acute LBP (<3 months) attending one of nine participating GPs. Baseline demographic, LBP classification [i.e. simple back ache (SBA), nerve root pain (NRP), serious spinal pathology (SSP)] and primary care management data were recorded over a three month period. Adherence and costs were estimated based on: medication prescription, referral for investigations, treatment or consultations, and wage replacement costs (time signed off work). For both SBA and NRP, medication prescriptions were consistent with European guideline recommendations, but not for referral for further treatment (39% of SBA patients were referred on first visit), secondary care (54% of NRP patients were referred on first visit), or discontinuation of work (50% NRP patients on first GP visit). The average total cost (direct and wage replacement) for a single episode of LBP over 12 weeks was 20,531 Euros (20,300-20,762). Direct costs accounted for 43% [8874.36 Euros, (8643.37-9105.37 Euros)] and wage replacement costs 57% (11,657 Euros). In conclusion, management of acute LBP in a cohort of GPs in Ireland was not consistent with European clinical guideline recommendations, and warrants higher levels of postgraduate education among GPs, as well as restructuring of primary care services, which should improve patient outcome and reduce costs. PMID- 17126047 TI - Relayed (13)C magnetization transfer: detection of malate dehydrogenase reaction in vivo. AB - Malate dehydrogenase catalyzes rapid interconversion between dilute metabolites oxaloacetate and malate. Both oxaloacetate and malate are below the detection threshold of in vivo MRS. Oxaloacetate is also in rapid exchange with aspartate catalyzed by aspartate aminotransferase, the latter metabolite is observable in vivo using (13)C MRS. We hypothesized that the rapid turnover of oxaloacetate can effectively relay perturbation of magnetization between malate and aspartate. Here, we report indirect observation of the malate dehydrogenase reaction by saturating malate C2 resonance at 71.2 ppm and detecting a reduced aspartate C2 signal at 53.2 ppm due to relayed magnetization transfer via oxaloacetate C2 at 201.3 ppm. Using this strategy the rate of the cerebral malate dehydrogenase reaction was determined to be 9+/-2 micromol/g wet weight/min (means+/-SD, n=5) at 11.7 Tesla in anesthetized adult rats infused with [1,6-(13)C(2)]glucose. PMID- 17126048 TI - Sensitivity enhancement in (13)C solid-state NMR of protein microcrystals by use of paramagnetic metal ions for optimizing (1)H T(1) relaxation. AB - We discuss a simple approach to enhance sensitivity for (13)C high-resolution solid-state NMR for proteins in microcrystals by reducing (1)H T(1) relaxation times with paramagnetic relaxation reagents. It was shown that (1)H T(1) values can be reduced from 0.4-0.8s to 60-70 ms for ubiquitin and lysozyme in D(2)O in the presence of 10 mM Cu(II)Na(2)EDTA without substantial degradation of the resolution in (13)C CPMAS spectra. Faster signal accumulation using the shorter (1)H T(1) attained by paramagnetic doping provided sensitivity enhancements of 1.4-2.9 for these proteins, reducing the experimental time for a given signal-to noise ratio by a factor of 2.0-8.4. This approach presented here is likely to be applicable to various other proteins in order to enhance sensitivity in (13)C high-resolution solid-state NMR spectroscopy. PMID- 17126049 TI - SOGGY: solvent-optimized double gradient spectroscopy for water suppression. A comparison with some existing techniques. AB - Excitation sculpting, a general method to suppress unwanted magnetization while controlling the phase of the retained signal [T.L. Hwang, A.J. Shaka, Water suppression that works. Excitation sculpting using arbitrary waveforms and pulsed field gradients, J. Magn. Reson. Ser. A 112 (1995) 275-279] is a highly effective method of water suppression for both biological and small molecule NMR spectroscopy. In excitation sculpting, a double pulsed field gradient spin echo forms the core of the sequence and pairing a low-power soft 180 degrees (-x) pulse with a high-power 180 degrees (x) all resonances except the water are flipped and retained, while the water peak is attenuated. By replacing the hard 180 degrees pulse in the double echo with a new phase-alternating composite pulse, broadband and adjustable excitation of large bandwidths with simultaneous high water suppression is obtained. This "Solvent-Optimized Gradient-Gradient Spectroscopy" (SOGGY) sequence is a reliable workhorse method for a wide range of practical situations in NMR spectroscopy, optimizing both solute sensitivity and water suppression. PMID- 17126050 TI - Interstitial deletion of 1q42.13-q43 with Duane retraction syndrome. AB - Duane retraction syndrome is a congenital eye movement disorder characterized by failure of the sixth cranial nerve or nucleus to develop properly, resulting in restriction of abduction, narrowing of the palpebral fissure, and retraction of the globe on attempted adduction and/or restricted adduction. The gene loci of Duane syndrome have been mapped on 8q and 2q, indicating a heterogeneity. We report here a patient with interstitial deletion of 1q with Duane syndrome. The karyotype of our patient suggests another possible locus of the Duane syndrome, and the mapped genes around the deleted region, 1q42.13-43, contain possible candidate genes such as a homeobox gene. However, further clinical descriptions of patients with Duane syndrome and genetic investigations of the deleted regions are needed for a more accurate delineation. PMID- 17126051 TI - Temporary incyclotorsion following surgical correction of bilateral superior oblique palsy. AB - We report three patients with acquired bilateral superior oblique paresis who had bilateral modified Harada-Ito procedures, which resulted in significant incyclotorsion (ranging from 6 degrees to 19 degrees ) on the first day postoperatively. For Case 1 this was the only procedure, while Cases 2 and 3 also underwent simultaneous bilateral inferior oblique weakening. This overcorrection decreased spontaneously over the following 8 months to leave all patients asymptomatic. One patient returned 8 years following the initial surgery with symptoms of diplopia and recurrence of excyclotorsion. PMID- 17126052 TI - Sulfur-dioxide exposure increases TRPV1-mediated responses in nodose ganglia cells and augments cough in guinea pigs. AB - The objective of the present experiments was to study the effects of pulmonary inflammation induced by subacute Sulfur-dioxide (SO(2)) exposure on capsaicin induced responses in isolated primary vagal sensory neurons and cough. Additionally, we examined the effects of SO(2) exposure on respiratory function and lung histology. All experiments were conducted 24 h after 4 days of subacute SO(2) (1000 ppm, 3 h/day for 4 days) exposure. In in vitro experiments, intracellular Ca(2+) concentrations were measured in single nodose ganglia cells isolated from SO(2) treated and control guinea pigs, using a fluorescence-based methodology. In nodose ganglia cells from SO(2)-exposed animals, intracellular Ca(2+) responses evoked by capsaicin (1 x 10(-7) and 1 x 10(-6) M) were significantly augmented (87% and 59%, respectively) compared to nodose ganglia from control animals. In vivo experiments, cough responses induced by a submaximal dose of aerosolized capsaicin (30 microM) were increased approximately 50% in SO(2) exposed animals compared to control animals. The enhanced cough response produced by SO(2) was inhibited by the corticosteroid, dexamethasone (10 mg/kg, p.o. b.i.d for 4 days and 10 mg/kg, p.o. once on day 5). In separate experiments, guinea pigs exposed to SO(2) displayed a decrease in respiratory frequency and minute ventilation and an increase in enhanced pause (PenH), a surrogate measure for pulmonary obstruction. Associated with the SO(2)-induced increase in cough and changes in respiratory parameters was an increase in BAL neutrophils. BAL neutrophil counts were 5+/-4 and 691+/-141 cells x 10(3)/ml for air and SO(2)-exposed animals, respectively. The neutrophillic inflammation induced by SO(2) was attenuated by dexamethasone treatment. Finally, staining for collagen, smooth muscle and goblet cells showed inflammation, remodeling and goblet cell metaphasia in the SO(2)-exposed animals. Our results demonstrate that SO(2) exposure enhances TRPV1 receptor function at the level of the nodose ganglia. This effect occurs in parallel with an increase sensitivity of the cough response to capsaicin. PMID- 17126053 TI - Poster discussion: summary. AB - The Fourth International Cough Symposium took place in London between the 29th of June and 1st of July 2006. There were overall 22 posters presented during the meeting. These posters were divided into the following sections: methods of cough investigations, definitions of cough, receptor mechanisms, neural pathways, animal experiments and clinical aspects of cough. This review will focus on the discussions related to the posters. PMID- 17126054 TI - Recombinant protein production in an Escherichia coli reduced genome strain. AB - Recently, efforts have been made to improve the properties of Escherichia coli as a recombinant host by 'genomic surgery'-deleting large segments of the E. coli K12 MG1655 genome without scars. These excised segments included K-islands, which contain a high proportion of transposons, insertion sequences, cryptic phage, damaged, and unknown-function genes. The resulting multiple-deletion strain, designated E. coli MDS40, has a 14% (about 700 genes) smaller genome than the parent strain, E. coli MG1655. The multiple-deletion and parent E. coli strains were cultured in fed-batch fermenters to high cell densities on minimal medium to simulate industrial conditions for evaluating growth and recombinant protein production characteristics. Recombinant protein production and by-product levels were quantified at different controlled growth rates. These results indicate that the multiple-deletion strain's growth behavior and recombinant protein productivity closely matched the parent stain. Thus, the multiple-deletion strain E. coli MDS40 provides a suitable foundation for further genomic reduction. PMID- 17126055 TI - Endoneural metastasis of the sciatic nerve disclosing the relapse of a renal carcinoma, four years after its surgical treatment. AB - METHODS: To report on an exceptional case of renal metastasis. Indeed, endoneural metastases of peripheral nerves are thought to be exceptional, and such an event has not yet been ascribed to a renal carcinoma. RESULTS: We report on a sciatica leading to the discovery of a tumour of the right thigh which proved to be an endoneural metastasis of the sciatic nerve from a renal carcinoma surgically removed 4 years before. CONCLUSION: A thorough palpation of peripheral nerve can help recognise such metastases in patients previously diagnosed with renal carcinoma. PMID- 17126056 TI - Development of recurrent pseudoseptic arthritis in a patient with rheumatoid arthritis receiving TNF-alpha blocker. PMID- 17126057 TI - Contribution of pharmacoepidemiological studies to the evaluation of treatment related adverse events. PMID- 17126058 TI - Symptomatic slow-acting drugs for osteoarthritis: what are the facts? AB - The term "symptomatic slow-acting drugs for osteoarthritis" (SySADOA) was coined more than a decade ago to designate medications and/or nutritional supplements used to alleviate the manifestations of osteoarthritis in the long-term. Their efficacy has always been a focus of considerable skepticism. However, a critical reappraisal of the available data, which include results of carefully designed clinical trials conducted in accordance with Good Clinical Practice guidelines, strongly suggests a therapeutic effect. The effects of SySADOA need to be determined based, in particular, on treatment objectives (symptom relief, decreased use of nonsteroidal antiinflammatory drugs and other conventional agents, decreased radiographic progression, and decreased use of joint replacement surgery). In addition, the characteristics of the patients who are most likely to benefit from SySADOA need to be identified. PMID- 17126059 TI - Bone density in young males with recently diagnosed inflammatory bowel disease. AB - OBJECTIVES: Patients with inflammatory bowel disease are at increased risk of developing osteopenia and osteoporosis. Our study was designed to determine the degree of decreased bone density in steroid naive young male patients with inflammatory bowel disease and to unmask possible risk factors. METHODS: Before the initiation of any treatment in young male patients aged 26 +/- 4.8 years with inflammatory bowel disease, ultrasound bone density measurement at the right calcaneous was performed using a Lunar Achilles plus device. Stiffness Index and T-score were measured. We also performed an ultrasound bone density measurement at right calcaneous in healthy age- and sex-matched controls. RESULTS: Nine out of 32 patients with inflammatory bowel disease had osteopenia or osteoporosis (approximately 28%). Of controls, two individuals had osteopenia (approximately 7%). There was a positive correlation between T-score and body mass index, but not between T-score and age in patients with inflammatory bowel disease. There was a statistically significant difference in T-score between patients with disease duration>6 months and those with disease duration<6 months (P=0.032), but not between the patients with Crohn's disease compared with the patients with ulcerative colitis. CONCLUSION: Steroid naive young male patients with inflammatory bowel disease have lower bone density values than healthy controls. According to our findings, duration of disease above 6 months and low body mass index are major risk factors for low bone density in these patients. Bone density measurement should be performed in all patients with inflammatory bowel disease in an early stage of the disease. PMID- 17126060 TI - Development and testing of a specific quality-of-life questionnaire for knee and hip osteoarthritis: OAKHQOL (OsteoArthritis of Knee Hip Quality Of Life). AB - OBJECTIVE: To design a quality-of-life (QOL) instrument specific for patients with knee or hip osteoarthritis and to assess its validity and reproducibility. METHODS: One-on-one or group interviews were conducted with 79 patients and 28 healthcare professionals. Of the 80 potential items identified from the results, 46 were selected based on their content and were used to develop version 1 of the OsteoArthritis of Knee and Hip Quality of Life Scale (OAKHQOL). The psychometric characteristics of the scale were evaluated in patients who met Altman's criteria for knee or hip osteoarthritis. RESULTS: Based on the results of psychometric analyses in 263 patients, three items were excluded, leaving 43 items in the final version (2.3) of the OAKHQOL. Principal components analysis identified four domains: physical activities, mental health, social functioning, and social support. A pain domain was individualized later. Construct validity, reproducibility, and discriminating power of the domains were satisfactory. Standardized response means after joint replacement surgery were close to 1 for the pain and physical functioning domains and equal to 0.7 for the mental health domain, indicating good sensitivity to change. CONCLUSIONS: The OAKHQOL is the first QOL tool specifically dedicated to lower-limb osteoarthritis. It captures specific aspects of QOL in patients with knee or hip osteoarthritis and exhibits psychometric properties consistent with use in longitudinal studies. PMID- 17126061 TI - Comparison of linear accelerations from three measurement systems during "reach & grasp". AB - Given the increased use of accelerometers in movement analysis, validation of such inertial sensors against conventional 3D camera systems and performance comparisons of different sensors have become important topics in biomechanics. This paper evaluates and compares linear acceleration trajectories obtained from two different 3D accelerometers and derived from Vicon position data for an upper limb "reach & grasp" task. Overall, good correspondence between the three measurement systems was obtained. Sources of error are discussed. PMID- 17126062 TI - Metrological validation for 3D modeling of dental plaster casts. AB - The contribution of this paper is twofold: (1) it presents an automatic 3D modeling technique and (2) it advances a procedure for its metrological evaluation in the context of a medical application, the 3D modeling of dental plaster casts. The motivation for this work is the creation of a "virtual gypsotheque" where cumbersome dental plaster casts can be replaced by numerical 3D models, thereby alleviating storage and access problems and allowing dentists and orthodontists the use of novel and unprecedented software tools for their medical evaluations. Modeling free-form surfaces of anatomical interest is an intriguing mixture of open issues concerning 3D modeling, geometrical metrology, and medicine. Of general interest is both the fact that a widespread use of 3D modeling in non-engineering applications requires automatic procedures of the kind presented in this work and the adopted validation paradigm for free-form surfaces, rather useful for practical purposes. In this latter respect, the metrological analysis we advance is the first seminal attempt in the field of 3D modeling and can be readily extended to contexts other than the medical one discussed in this paper. PMID- 17126064 TI - Semi-automatic level-set based segmentation and stenosis quantification of the internal carotid artery in 3D CTA data sets. AB - We present a new level-set based method to segment and quantify stenosed internal carotid arteries (ICAs) in 3D contrast-enhanced computed tomography angiography (CTA). Within these data sets it is a difficult task to evaluate the degree of stenoses deterministically even for the experienced physician because the actual vessel lumen is hardly distinguishable from calcified plaque and there is no sharp border between lumen and arterial wall. According to our knowledge no commercially available software package allows the detection of the boundary between lumen and plaque components. Therefore in the clinical environment physicians have to perform the evaluation manually. This approach suffers from both intra- and inter-observer variability. The limitation of the manual approach requires the development of a semi-automatic method that is able to achieve deterministic segmentation results of the internal carotid artery via level-set techniques. With the new method different kinds of plaques were almost completely excluded from the segmented regions. For an objective evaluation we also studied the method's performance with four different phantom data sets for which the ground truth of the degree of stenosis was known a priori. Finally, we applied the method to 10 ICAs and compared the obtained segmentations with manual measurements of three physicians. PMID- 17126065 TI - Automatic rib segmentation and labeling in computed tomography scans using a general framework for detection, recognition and segmentation of objects in volumetric data. AB - A system for automatic segmentation and labeling of the complete rib cage in chest CT scans is presented. The method uses a general framework for automatic detection, recognition and segmentation of objects in three-dimensional medical images. The framework consists of five stages: (1) detection of relevant image structures, (2) construction of image primitives, (3) classification of the primitives, (4) grouping and recognition of classified primitives and (5) full segmentation based on the obtained groups. For this application, first 1D ridges are extracted in 3D data. Then, primitives in the form of line elements are constructed from the ridge voxels. Next a classifier is trained to classify the primitives in foreground (ribs) and background. In the grouping stage centerlines are formed from the foreground primitives and rib numbers are assigned to the centerlines. In the final segmentation stage, the centerlines act as initialization for a seeded region growing algorithm. The method is tested on 20 CT-scans. Of the primitives, 97.5% is classified correctly (sensitivity is 96.8%, specificity is 97.8%). After grouping, 98.4% of the ribs are recognized. The final segmentation is qualitatively evaluated and is very accurate for over 80% of all ribs, with slight errors otherwise. PMID- 17126063 TI - Retinotopic organization of primary visual cortex in glaucoma: Comparing fMRI measurements of cortical function with visual field loss. AB - Primary open angle glaucoma (POAG) is a progressive optic neuropathy characterized by retinal ganglion cell loss. Experimental primate glaucoma indicates neuronal degeneration of the lateral geniculate nucleus (LGN) and activity changes in the visual cortex (V1). Neuronal degeneration has also been shown in a post-mortem human study of the optic nerve, LGN and visual cortex. Functional magnetic resonance imaging (fMRI), a non-invasive means of inferring function-specific neuronal activity, provides an opportunity to evaluate glaucomatous changes in neuronal activity throughout the visual pathway in vivo. The purpose of this study is to demonstrate that the relationship between visual field loss in human POAG and the functional organization of V1 can be measured using novel fMRI analysis methods. Visual field defects were measured using standard automated perimetry (SAP). A retinotopic map of visual space was obtained for V1, and the retinotopy data was fit with a template. The template was used to project regions within the visual field onto a flattened representation of V1. Viewing through the glaucomatous vs. fellow eye was compared by alternately presenting each eye with a scotoma-mapping stimulus. The resulting blood oxygen level dependent (BOLD) fMRI response was compared to interocular differences in thresholds for corresponding regions of the visual field. The spatial pattern of activity observed in the flattened representation agreed with the pattern of visual field loss. Furthermore, the amplitude of the BOLD response was correlated on a pointwise basis with the difference in sensitivity thresholds between the glaucomatous and fellow eyes (r = 0.53, p < 0.0001). The BOLD signal in human V1 is altered for POAG patients in a manner consistent with the loss of visual function. FMRI of visual brain areas is a potential means for quantifying glaucomatous changes in neuronal activity. This should enhance our understanding of glaucoma, and could lead to new diagnostic techniques and therapies. PMID- 17126066 TI - Strategy for genotoxicity testing: hazard identification and risk assessment in relation to in vitro testing. AB - This report summarizes the proceedings of the September 9-10, 2005 meeting of the Expert Working Group on Hazard Identification and Risk Assessment in Relation to In Vitro Testing, part of an initiative on genetic toxicology. The objective of the Working Group was to develop recommendations for interpretation of results from tests commonly included in regulatory genetic toxicology test batteries, and to propose an appropriate strategy for follow-up testing when positive in vitro results were obtained in these assays. The Group noted the high frequency of positive in vitro findings in the genotoxicity test batteries with agents found not to be carcinogenic and thought not to pose a carcinogenic health hazard to humans. The Group agreed that a set of consensus principles for appropriate interpretation and follow-up testing when initial in vitro tests are positive was needed. Current differences in emphasis and policy among different regulatory agencies were recognized as a basis of this need. Using a consensus process among a balanced group of recognized international authorities from industry, government, and academia, it was agreed that a strategy based on these principles should include guidance on: (1) interpretation of initial results in the "core" test battery; (2) criteria for determining when follow-up testing is needed; (3) criteria for selecting appropriate follow-up tests; (4) definition of when the evidence is sufficient to define the mode of action and the relevance to human exposure; and (5) definition of approaches to evaluate the degree of health risk under conditions of exposure of the species of concern (generally the human). A framework for addressing these issues was discussed, and a general "decision tree" was developed that included criteria for assessing the need for further testing, selecting appropriate follow-up tests, and determining a sufficient weight of evidence to attribute a level of risk and stop testing. The discussion included case studies based on actual test results that illustrated common situations encountered, and consensus opinions were developed based on group analysis of these cases. The Working Group defined circumstances in which the pattern and magnitude of positive results was such that there was very low or no concern (e.g., non-reproducible or marginal responses), and no further testing would be needed. This included a discussion of the importance of the use of historical control data. The criteria for determining when follow-up testing is needed included factors, such as evidence of reproducibility, level of cytotoxicity at which an increased DNA damage or mutation frequency is observed, relationship of results to the historical control range of values, and total weight of evidence across assays. When the initial battery is negative, further testing might be required based on information from the published literature, structure activity considerations, or the potential for significant human metabolites not generated in the test systems. Additional testing might also be needed retrospectively when increase in tumors or evidence of pre-neoplastic change is seen. When follow-up testing is needed, it should be based on knowledge about the mode of action, based on reports in the literature or learned from the nature of the responses observed in the initial tests. The initial findings, and available information about the biochemical and pharmacological nature of the agent, are generally sufficient to conclude that the responses observed are consistent with certain molecular mechanisms and inconsistent with others. Follow up tests should be sensitive to the types of genetic damage known to be capable of inducing the response observed initially. It was recognized that genotoxic events might arise from processes other than direct reactivity with DNA, that these mechanisms may have a non-linear, or threshold, dose-response relationship, and that in such cases it may be possible to determine an exposure level below which there is negligible concern about an effect due to human exposures. When a test result is clearly positive, consideration of relevance to human health includes whether other assays for the same endpoint support the results observed, whether the mode or mechanism of action is relevant to the human, and - most importantly - whether the effect observed is likely to occur in vivo at concentrations expected as a result of human exposure. Although general principles were agreed upon, time did not permit the development of recommendations for the selection of specific tests beyond those commonly employed in initial test batteries. PMID- 17126067 TI - The spectral studies on the effect of Glu 101 to the metal binding characteristic of Euplotes octocarinatus centrin. AB - Glu is highly conserved as the first amino acid of E-helix of the EF-hand protein. In this paper, Glu 101, the first amino acid of E-helix of the third EF hand motif in Euplotes octocarinatus centrin (EoCen) was mutated to be Lys by the method of site direct mutation. Tb3+ and TNS were used as fluorescence probes in the study of the effect of this mutation to the metal binding characteristic of EoCen by fluorescence spectra. Results indicate that compared with EoCen, the mutation protein (E101K) displays a different Tb3+ binding characteristic and an increased hydrophobic exposure surface. Polyacrylamide gels electrophoresis indicated that the electrophoretic mobilities of EoCen and E101K are distinctly different. It can be deduced that the conformation of EoCen has been altered by this mutation. The general conditional binding constant of Tb3+ to the three loops of EF-hand sites I-III in E101K was calculated to be (5.64+/-0.57)x10(5)M( 1) according to the modified equation of the single binding process. PMID- 17126068 TI - Electronic absorption study on acid-base equilibria for some keto and thioketo pyrimidine derivatives. Experimental and theoretical evidence of enolization and solute-solvent interactions. AB - The UV-vis spectra of recently synthesized 1-amino-5-benzoyl-4-phenyl-1H pyrimidine-2-one, (I), and 1-amino-5-benzoyl-4-phenyl-1H-pyrimidine-2-thione (II), were studied in aqueous methanol (5%, v/v, methanol) and pure methanol. The nature of the electronic transitions and the role of carbonyl oxygen of I and thiocarbonyl sulfur of II in the behavior of the observed UV-vis spectra were discussed. The carbonyl group at position 2 of I and the thiocarbonyl group of II were found to be enolized instead of protonation. Quantum chemical calculations showed agreement with the experimental evidence. However, the carbonyl group of the benzoyl moiety at position 5 of both compounds underwent neither enolization nor protonation. Acid-base equilibria of the compounds against varying pH have been examined in detail. The pKa values of all related equilibria were determined at room temperature and an ionic strength of 0.10 M from the pH-dependence of the absorbance values using the Henderson-Haselbalch equation and graphical logarithmic analysis. The mean acidity constants for the protonated forms of the compounds were determined as pKa1=4.214 and pKa2=6.678 for I and pKa1=3.739 and pKa2=6.258 for II. The mean acidity constants (pKa3) for the enol form of I and the thioenol form of II were determined as 11.278 and 11.063, respectively. The preferred dissociation mechanisms were discussed based on the data of UV-vis spectroscopy and a mechanism was proposed for each compound. The formation of intramolecular and intermolecular hydrogen bonding were found with I but not with II. The intramolecular bonding stabilizing the enol form was favoured at pH values corresponding to pKa1 and above. On the other hand, the intermolecular hydrogen bonding stabilizing the free form of the carbonyl group was favoured at all pH values. PMID- 17126069 TI - Investigating risk exposure in e-health systems. AB - PURPOSE: Health managers, administrators and health practitioners now face new challenges due to the increasing dependency being placed on electronic health information systems. This paper focuses on Electronic Health Records for determining the critical attributes for e-health system development. The proposed QUiPS model aims to provide a framework for building trustworthy solutions by identifying the pertinent issues needed to determine the risk exposure with a given system. APPROACH: To produce dependable, low risk and viable IT solutions, each critical attribute needs to be specifically addressed and prioritized. It is shown how these attributes possess a number of interdependencies making the analysis and prioritization tasks complex and hence, in practice, often incomplete. Two Australian case studies are presented that access enterprise level applications of live health records where these risk based techniques have been applied. RESULTS: The value and the shortcomings of taking a risk based approach to developing and deploying electronic health information systems that are safe and secure, is evaluated. The case studies presented indicate that traditional methods used to derive the requirements are often inadequate and the risks that are faced in ensuring a safe and secure system are highly application dependent and dynamic. CONCLUSIONS: Convergence towards a viable universal solution for our electronic health records is not imminent and trust in e-health is fragile. Policies that data custodians follow need to be flexible and updated on a regular basis. Technological solutions are at best a stop gap to avoid the common hazards associated with access control and secure messaging. A wider range of analysis techniques to determine the key issues for a dependable health information system can derive longer term sustainable solutions. PMID- 17126070 TI - Evoked gamma oscillations in human scalp EEG are test-retest reliable. AB - OBJECTIVE: Evoked, phase-locked gamma oscillations of the electroencephalogram (EEG) have been demonstrated to be modulated by both bottom-up as well as top down factors. However, to date the test-retest reliability of these oscillations has not been studied systematically. METHODS: We recorded EEG activity of 12 healthy volunteers in response to stimuli of different sizes. Each participant took part in two sessions separated by two weeks in time. To obtain an estimate of the reliability of evoked gamma band responses (GBRs), we compared frequency and magnitude of phase-locked EEG oscillations between sessions. RESULTS: In response to large stimuli magnitude and frequency of the evoked GBR yielded significant reliability. However, this was not the case for stimuli which were too small to evoke detectable GBRs. CONCLUSIONS: The results are in accordance with studies demonstrating a dependence of gamma oscillations on stimulus parameters. SIGNIFICANCE: The current findings suggest that using appropriate stimulation, the evoked gamma response has sufficient test-retest reliability for use in assessing clinical changes in neurophysiological status. PMID- 17126071 TI - Localizing value of scalp EEG spikes: a simultaneous scalp and intracranial study. AB - OBJECTIVE: To determine the relationship between cortical origins of interictal and ictal EEG discharges in patients with temporal lobe epilepsy. METHODS: Simultaneous cortical and scalp EEG recordings were obtained from six patients with temporal lobe epilepsy. Subdural electrode contacts active at seizure onset and when scalp ictal rhythms became evident were identified. Similarly, cortical substrates of scalp EEG spikes were identified at spike peak and at the initial rising phase of the potential. RESULTS: Intracranial seizure onsets were commonly focal and involved only a few electrode contacts, as opposed to scalp ictal rhythms, which required synchronous activation of multiple electrode contacts. At the peak of scalp spikes, multiple electrode contacts were similarly active. However, at spike onset, cortical substrates were more discrete and commonly involved electrodes similar to that of seizure onsets. CONCLUSIONS: Scalp EEG ictal rhythms and the peak of a scalp spike may poorly localize the epileptogenic focus because of propagation. Cortical source area at scalp spike onset is more discrete, however, and the seizure onset zone often lies within this area. SIGNIFICANCE: Analysis of scalp spikes, such as source modeling, at their initial rising phase might provide useful localizing information about seizure origins in the same patient. PMID- 17126073 TI - Effects of aldosterone receptor blockade in patients with mild-moderate heart failure taking a beta-blocker. AB - AIMS: Spironolactone improves prognosis in severe heart failure (HF). We investigated its effects in patients with mild-moderate HF treated with an ACE inhibitor and beta-blocker. METHODS AND RESULTS: Randomised, double-blind, parallel-group, 3-month comparison of placebo and spironolactone (25 mg daily) in 40 patients in New York Heart Association (NYHA) class I (20%), II (70%) or III (10%), with a left ventricular ejection fraction of <40%. The mean (standard error) changes from baseline in the spironolactone and placebo groups were, respectively: i) B-type natriuretic peptide (BNP) -53.4(22.2) pg/mL and +3.3(12.1) pg/mL, P=0.04, ii) pro-collagen type III N-terminal amino peptide (PIIINP) -0.6(0.2) micromol/L and +0.02(0.2) micromol/L, P=0.02 and iii) creatinine +10.7(3.2) micromol/L and -0.3(2.6) micromol/L, P=0.01. Compared with placebo, spironolactone therapy was associated with a reduction in self-reported health-related quality of life: change in visual analog score: -6 (3) vs. +6 (4); P=0.01. No differences were observed on other biochemical, neurohumoral, exercise and autonomic function assessments. CONCLUSION: In patients with mild-moderate HF, spironolactone reduced neurohumoral activation (BNP) and a marker of collagen turnover (PIIINP) but impaired renal function and quality of life. The benefit risk ratio of aldosterone blockade in mild HF is uncertain and requires clarification in a large randomised trial. PMID- 17126075 TI - Optimization of culture conditions for a synthetic gene expression in Escherichia coli using response surface methodology: the case of human interferon beta. AB - A human interferon beta (hINF-beta) synthetic gene was optimized and expressed in Escherichia coli BL21-SI using a vector with the T7 promoter. To determine the best culture conditions such as culture medium, temperature, cell density and inducer concentration, we used the response surface methodology and a Box-Behnken design to get the highest hINF-beta production. The maximum hINF-beta production of 61 mg l(-1) was attained using minimum medium and the following predicted optimal conditions: temperature of 32.5 degrees C, cell density of 0.64, and inducer concentration of 0.30 M NaCl. This is the first report showing the successful performance of the BL21-SI system in a minimum medium. The response surface methodology is effective for the optimization of recombinant protein production using synthetic genes. PMID- 17126076 TI - Homologs of the sexually induced gene 1 (sig1) product constitute the stramenopile mastigonemes. AB - The tripartite tubular mastigoneme on the anterior flagellum is a morphological feature that characterizes the stramenopiles. Mastigonemes are significant and potentially informative structures not only from the viewpoint of systematics, but also of cell biology. Nevertheless, few biochemical studies have been reported on stramenopile mastigonemes. The flagella of Scytosiphon lomentaria (Phaeophyceae) were successfully isolated and analyzed using SDS-PAGE followed by protein sequencing. The partial amino acid sequence of one flagellar protein (115kDa) showed high similarity with the sexually induced gene 1 (sig1) product of centric diatoms. A polyclonal antibody against the 115-kDa protein reacted not only to the shaft of mastigonemes in Scytosiphon lomentaria, but also another distinctly different stramenopile flagellate, Sulcochrysis biplastida (Dictyochophyceae). Therefore, we propose that the 115-kDa protein (i.e. Sig1 homologs) is a constituent of the tubular shaft of the mastigoneme. PMID- 17126077 TI - Analysis of physical match performance in English Premier League soccer referees with particular reference to first half and player work rates. AB - The aim of the present study was to examine the influence of first half activity, overall match intensity and seasonal variation on the physical match performances of English Premier League football referees. Match analysis data was collected using the Prozone match analysis system from 19 full-time professional referees during a total of 254 matches in the 2004-2005 season. Physical match performances were classified into three separate categories: 1, total distance covered (TD); 2, high-intensity running distance (running speed>5.5m/s, HIR); 3, average distance from infringements (DI). Using these match activity variables the influence of first half TD and HIR distances on second half activities and also the influence of players' match activities upon the referees' physical match performances were examined. The main finding of the present study was that the physical match performances of the referees were partly related to those of the players, in that the referees' HIR correlated with players' HIR (r=0.43, p<0.0001, n=212). Furthermore, first half TD and HIR distances were found to be related to second half coverage in referees (r=0.47 and r=0.52, respectively, p<0.001, n=254). These results demonstrate a need to assess the overall match intensity prior to examining the physical match performance of the referee. Further examination is required as to whether reduced physical performances in the second half of matches are a consequence of referee fatigue, tactical strategies on behalf of the referee or reduced player match activities resulting in a slower tempo of match. PMID- 17126078 TI - Preparation for cancer care: perceptions of newly qualified health care professionals. AB - The present paper is derived from a larger survey which examined the perceptions of recently qualified health care professionals' experience on evidence-based practice, team working and cancer care. This study reports solely on the findings relating to cancer care. The perceptions of recently qualified professionals in relation to their initial educational input on issues such as confidence, anxiety, communication skills and practice in cancer care as well as adequacy of support, professional supervision and use of reflection were gathered using a cross-sectional postal survey design. A total of 50 graduates from each professional category in nursing, occupational therapy, physiotherapy, and social work were sampled yielding a total sample of 200. Eighty-five questionnaires were returned yielding a response rate of 43%. Twenty-eight (33%) respondents stated that they were currently involved in working with people with cancer. These were as follows: 5 nurses, 8 physiotherapists, 9 occupational therapists and 6 social workers. Despite the low response rate, the findings suggest that health care professionals' educational input and experiences of working with people with cancer were overall positive; for example, in the respondents' confidence, communication skills, decrease in anxiety and application of knowledge gained in classroom to professional practice. Moreover, most respondents learnt about caring for cancer patients through practice rather than classroom teaching. A high percentage (i.e. 64%;18) across all groups felt supported when caring for people with cancer and reported receiving professional supervision as well as being able to actively reflect on their practice. The implications for education and practice were discussed particularly as there have been few studies conducted in relation to the specific needs and collaborative learning of these health care professional groups. PMID- 17126079 TI - Link test--A statistical method for finding prostate cancer biomarkers. AB - We present a new method, link-test, to select prostate cancer biomarkers from SELDI mass spectrometry and microarray data sets. Biomarkers selected by link test are supported by data sets from both mRNA and protein levels, and therefore results in improved robustness. Link-test determines the level of significance of the association between a microarray marker and a specific mass spectrum marker by constructing background mass spectra distributions estimated by all human protein sequences in the SWISS-PROT database. The data set consist of both microarray and mass spectrometry data from prostate cancer patients and healthy controls. A list of statistically justified prostate cancer biomarkers is reported by link-test. Cross-validation results show high prediction accuracy using the identified biomarker panel. We also employ a text-mining approach with OMIM database to validate the cancer biomarkers. The study with link-test represents one of the first cross-platform studies of cancer biomarkers. PMID- 17126080 TI - IL-1 beta stimulates divergent upper and lower airway epithelial cell CCL5 secretion. AB - Direct infection of respiratory epithelium induces chemokine secretion and upregulates cytokine networks, which are central in regulating inflammation. IL 1beta may have a pivotal role in such networks. Differential control of chemokine secretion within specific airway regions, which have distinct roles in immunity, is not well characterized. We investigated IL-1beta-induced CXCL8 and CCL5 secretion from primary normal human bronchial and small airway epithelial cells, and the alveolar cell line A549. CXCL8 was secreted by all cells, but only lower airway cells secreted CCL5. IL-1beta induced nuclear translocation of NF-kappaB (p50, p65 and c-Rel subunits), NF-IL-6 and AP-1, each with distinct kinetics. This was associated with high level CCL5 promoter activation, via transcription factor binding to multiple regions, including NF-kappaB, AP-1 and NF-IL-6 sites. The IL-1-related cytokine IL-18 did not drive or modulate IL-1beta-induced CXCL8 or CCL5 secretion. In summary, IL-1beta, but not IL-18, induces transcription dependent lower airway epithelial cell-specific CCL5 secretion. Differential chemokine secretion may have profound effects on local leukocyte influx within upper or lower airways exposed to airway infection or environmental stimuli, which might then require different anti-inflammatory strategies. PMID- 17126081 TI - Mathematical modeling as accounting: predicting the fate of serum proteins and therapeutic monoclonal antibodies. AB - This article reviews current efforts to mathematically model the half lives of serum proteins, especially antibodies. While it is recognized that the neonatal Fc receptor, FcRn, is necessary for longer serum persistence of certain proteins, particularly the high abundance IgGs and albumin, it is not clear that it is sufficient to completely determine the half lives of these proteins. More specifically, it is unclear why the high avidity (bivalent), high affinity FcRn IgG interaction, with half saturation in the 10 to 100 nM range (at endosomal pH according to the currently proposed mechanism), would result in a salvage mechanism that saturates at serum concentrations in the 10 to 100 mg/ml range--a discrepancy of 4 to 5 orders of magnitude. Alternative explanations include the proposal that the very low affinity binding between FcRn and IgG at blood pH is also relevant to the salvage mechanism, and that factors in addition to FcRn binding modulate the maintenance and clearance of IgG from serum. PMID- 17126082 TI - famoses: a modular educational program for children with epilepsy and their parents. AB - BACKGROUND: It is known that most adult patients with epilepsy often have poor knowledge of their disorder, treatment options, and social and vocational consequences, despite the huge amount of information available. Being pressed for time, health care professionals often are not able to provide the repetitive counseling and instruction necessary to address epilepsy care adequately. Therefore educational programs are considered extremely important in filling the gap. METHOD: For German-speaking countries, two educational programs called famoses, modulares Schulungsprogramm Epilepsie fur Familien [modular service package epilepsy for families], were developed by a multidisciplinary group of neuropediatricians, psychologists, social workers, and educators. The aims of the famoses programs are to improve children's and parents' knowledge about epilepsy and to help patients of childhood age and their parents achieve a better understanding of their disease, gain more self-confidence, and reduce specific fears regarding epilepsy. famoses comprises two different educational programs: famoses for kids with epilepsy within the developmental period of ages 7 to 13, and famoses for parents or caretakers. The programs are designed for interactive small-group education. RESULTS: The child-centered educational program is based on a fictional story: The children are sailors on a virtual cruise, sailing from island to island, accompanied by educated trainers. On each island there is something new to discover about epilepsy. The parent (and caretaker)-centered educational program covers different topics-overview and content of the program, basic knowledge, diagnostics, therapy, prognosis and development, and living with epilepsy-within six modules. The program was implemented in Germany and Switzerland in the spring of 2005 and is now operating in different epilepsy centers. CONCLUSION: Within comprehensive therapeutic management of epilepsy, educational programs for patients, and for parents (caretakers) of children with epilepsy, are considered to be extremely important. Such educational programs have two main goals: to increase knowledge about the disorder, and to strengthen the patients' responsibility for themselves, with the consequence of living with as few limitations as possible. famoses is an effective component of this comprehensive care. The program has been operating successfully in different epilepsy centers in Germany and Switzerland since the spring of 2005. PMID- 17126083 TI - Mechanism of interaction between human 8-oxoguanine-DNA glycosylase and AP endonuclease. AB - Human 8-oxoguanine-DNA glycosylase (OGG1) is the main human base excision protein that removes a mutagenic lesion 8-oxoguanine (8-oxoG) from DNA. Since OGG1 has DNA glycosylase and weak abasic site (AP) lyase activities and is characterized by slow product release, turnover of the enzyme acting alone is low. Recently it was shown that human AP endonuclease (APE1) enhances the activity of OGG1. This enhancement was proposed to be passive, resulting from APE1 binding to or cleavage of AP sites after OGG1 dissociation. Here we present evidence that APE1 could actively displace OGG1 from its product, directly increasing the turnover of OGG1. We have observed that APE1 forms an electrophoretically detectable complex with OGG1 cross-linked to DNA by sodium borohydride. Using oligonucleotide substrates with a single 8-oxoG residue located in their 5' terminal, central or 3'-terminal part, we have demonstrated that OGG1 activity does not increase only for the first of these three substrates, indicating that APE1 interacts with the DNA stretch 5' to the bound OGG1 molecule. In kinetic experiments, APE1 enhanced the product release constant but not the rate constant of base excision by OGG1. Moreover, OGG1 bound to a tetrahydrofuran analog of an abasic site stimulated the activity of APE1 on this substrate. Using a concatemeric DNA substrate, we have shown that APE1 likely displaces OGG1 in a processive mode, with OGG1 remaining on DNA but sliding away in search for a new lesion. Altogether, our data support a model in which APE1 specifically recognizes an OGG1/DNA complex, distorts a stretch of DNA 5' to the OGG1 molecule, and actively displaces the glycosylase from the lesion. PMID- 17126084 TI - Characterisation of Ercc1 deficiency in the liver and in conditional Ercc1 deficient primary hepatocytes in vitro. AB - The ERCC1/XPF complex is responsible for incision at the 5' side of the lesion during nucleotide excision repair and is also involved in homologous recombination and interstrand cross-link repair. The aim of the current study was to set up a better model for examination of Ercc1 deficiency in the murine liver and to determine the DNA lesions responsible for the premature polyploidy observed. We used the Cre/lox system with an adenovirus carrying Cre recombinase to conditionally induce Ercc1 deficiency in murine hepatocytes in vitro. Increased levels of apoptosis were apparent in our Ercc1-deficient cultures, both spontaneously and after UV irradiation and oxidative DNA damage. Increased apoptosis was also observed in simple Ercc1-deficient livers and the time course of the development of polyploidy was characterised. Livers from simple Ercc1 knockout mice contained mitochondria with disrupted outer membranes. Lipid accumulation was observed in older Ercc1-deficient hepatocyte cultures and in young Ercc1-deficient and wild-type livers. Lipids disappeared from the wild-type livers with age, but persisted in Ercc1-deficient livers, suggesting that a reduced ability to repair oxidative DNA damage and a malfunction of oxidative pathways could be responsible for the Ercc1-deficient liver phenotype. Real-time RT-PCR was used to determine differences in expression of cell cycle regulation and survival genes between Ercc1-deficient and control livers. Higher mRNA levels of Igfbp2, a possible marker for polyploidy, and p21 were detected in Ercc1 deficient livers. The pro-apoptotic factor, Bax, showed increased levels of mRNA expression in young Ercc1-deficient livers. However, no elevation in the levels of reactive oxygen species, or of malondialdehyde DNA adducts, a product of oxidative DNA damage, were found in Ercc1-deficient liver and no elevated levels of genes involved in the oxidative damage response were seen. PMID- 17126085 TI - Molecularly imprinted polymer microspheres for solid-phase extraction of chloramphenicol residues in foods. AB - Preparation of molecularly imprinted polymer microspheres (MIPMs) for chloramphenicol (CAP) by aqueous suspension polymerization is reported for the first time in this study. The resulting MIPMs had the ability to specifically adsorb CAP, and the molecularly imprinted solid phase extraction (MISPE) based on the MIPMs was shown to be applicable for clean-up and preconcentration of trace CAP in milk and shrimp samples with high recoveries of 92.7% and 84.9%, respectively. Combined with MISPE, the conventional HPLC-UV analysis sensitivity for CAP in foods could be significantly increased. PMID- 17126086 TI - A sensitive assay for simultaneous determination of plasma concentrations of valganciclovir and its active metabolite ganciclovir by LC/MS/MS. AB - A protein precipitation, liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed and validated for the simultaneous determination of valganciclovir and its active metabolite ganciclovir in human plasma. The solvent system also served as a protein precipitation reagent. The chromatographic separation was achieved on an Aquasil C18 column (50 mm x 2.1mm, 5 microm). A linear gradient mobile phase between 0.02% formic acid and methanol was used. Detection was by positive ion electrospray tandem mass spectrometry on a Sciex API3000. The standard curves, which ranged from 4 to 512 ng/mL for valganciclovir and from 0.1 to 12.8 microg/mL for ganciclovir, were fitted to a 1/x weighted quadratic regression model. The method was proved to be accurate, specific and sensitive enough and was successfully applied to a pharmacokinetic study. PMID- 17126087 TI - Validation of high-performance liquid chromatography assay for quantification of formoterol in urine samples after inhalation using UV detection technique. AB - A novel high-performance liquid chromatography (HPLC) assay for the estimation of formoterol in urine samples was developed and validated. A solid phase extraction (SPE) using Oasis HLB was optimised to isolate formoterol from a urine matrix followed by HPLC with UV detection. This extraction procedure concentrated the final analyte forty times so that UV detection can be used to determine even a low concentration of formoterol in urine samples. The urinary assay was performed in accordance with FDA and ICH regulations for the validation of bioanalytical samples. The samples were injected onto a C18 Spherisorb (250 mm x 4.6 mm x 5 microm) analytical column maintained at 30 degrees C. The mobile phase consisted of 5 mM of potassium dihydrogen orthophosphate buffer (adjusted to pH 3 with ortho phosphoric acid):acetonitrile (ACN) (70:30, v/v), and the formoterol peak was detected at wavelength 214 nm. The extraction recovery of formoterol from the urine sample was >95%. The calibration curve was linear (r2=0.99) over formoterol concentrations ranging from 1.5 to 25 ng/mL (n=6). The method had an accuracy of >92% and intra and inter-day precision CV% of <3.9% and <2.2%, respectively, at three different concentrations low, medium and high (10, 15, 20 ng/mL). The limit of quantification (LOQ) for formoterol was found to be 1.50 ng/mL. The accuracy and precision at the LOQ level were 95% and %CV <3.7% (n=10), respectively. The method reported is simple, reliable, precise, and accurate and has the capacity to be used for determination of formoterol in urine samples. PMID- 17126088 TI - Resolution of protein-losing enteropathy and normalization of mesenteric Doppler flow with sildenafil after Fontan. AB - A 9.5-year-old girl after Fontan procedure for hypoplastic left heart syndrome had recurrent protein-losing enteropathy (PLE) develop 2 months after partial catheter closure of the Fontan fenestration. Despite satisfactory hemodynamic measurements under general anesthesia, we postulated that she suffered from increased pulmonary vascular reactivity and commenced her on Sildenafil treatment. After 6 weeks of oral Sildenafil treatment, her serum albumin and the fecal alpha-1-antitrypsin levels normalized, and her exercise tolerance had increased. There was also an improvement of the mesenteric arterial flow patterns on Doppler studies. Sildenafil should be considered in the treatment of PLE after the Fontan procedure. PMID- 17126089 TI - Severe superior vena cava syndrome after transvenous pacemaker implantation. AB - Superior vena cava syndrome is a rare complication after pacemaker implantation. The present report outlines how underestimation of clinically relevant symptoms of superior vena cava syndrome early after pacemaker implantation may result in severe complication with the need for major surgical intervention. Superior vena cava syndrome should be diagnosed early because immediate thrombolytic therapy is effective in the majority of patients and avoids the requirement for interventional or surgical efforts. PMID- 17126090 TI - Awake tracheobronchial dilation without the use of rigid bronchoscopy. AB - Benign tracheal strictures have been treated by repeated dilations using rigid bronchoscopy and general anesthesia. An alternative approach using conscious sedation, fluoroscopy, flexible bronchoscopy, and Savary-Gilliard esophageal dilators (Cook Endoscopy, Inc, Winston-Salem, NC) is reported. All patients undergoing dilation while awake had lesions due to inflammatory disease or lesions that were not amenable to resection. This method may be less traumatic than rigid bronchoscopy and can be accomplished on an outpatient basis without the use of general anesthesia. PMID- 17126091 TI - Folding valvuloplasty without leaflet resection: simplified method for mitral valve repair. AB - Standard repair techniques can prove challenging in patients with severe myxomatous mitral valve disease when there is a broad-based mid-portion of the posterior leaflet and small medial and lateral scallops. We describe a new surgical technique in which the mitral valve repair was accomplished by simple folding the prolapsed segment of the posterior leaflet and by insertion of the flexible annuloplasty ring. PMID- 17126093 TI - Thomas B. Ferguson Lecture. Coronary artery bypass grafting is still the best treatment for multivessel and left main disease, but patients need to know. PMID- 17126094 TI - Predictors of acute lung injury and severe hypoxemia in patients undergoing operative talc pleurodesis. AB - BACKGROUND: Acute lung injury (ALI) is a life-threatening complication of talc pleurodesis. This study defines characteristics that predispose patients to ALI and severe hypoxemia in patients after video-assisted thoracoscopic surgery (VATS) talc pleurodesis. METHODS: Charts of patients who underwent talc pleurodesis at Northwestern Memorial Hospital between January 1, 1997 and December 31, 2003 were retrospectively reviewed. We sought variables associated with the development of postoperative ALI or severe hypoxemia, defined as an increase in fraction of the inspired oxygen by more than 0.15 within 24 hours after the surgery. The analysis included 84 patients (58 women, 26 men) who underwent VATS talc pleurodesis for malignant (n = 74) or benign (n = 10) indications. RESULTS: ALI developed in 5 patients (5.9%), severe hypoxemia developed in 25 (29.8%), and 54 (64.3%) did not have postoperative complications. In multivariate analysis, the presence of peripheral edema before pleurodesis (p = 0.005), any preoperative requirement for supplemental oxygen (p = 0.032), and chemotherapy within 14 days before pleurodesis (p = 0.04) were identified as predictors of ALI or severe postoperative hypoxemia. CONCLUSIONS: Oxygen supplementation, recent chemotherapy, and presence of peripheral edema were independent predictors of a combined outcome of ALI or severe hypoxemia after VATS talc pleurodesis. Patients with these characteristics might be at risk for adverse outcomes of talc pleurodesis and should be considered for alternative therapy for their effusions. PMID- 17126095 TI - Increased risk of aspiration and pulmonary complications after lung resection in head and neck cancer patients. AB - BACKGROUND: Patients with head and neck cancers (HNCs) may have impaired deglutition and airway protection. The risk of pulmonary complications after lung resection has not been evaluated in HNC patients. We therefore reviewed the risk of aspiration-related pulmonary complications in HNC patients after pulmonary resection. METHODS: Retrospective review of a prospectively collected database from a single institution identified 1633 patients who underwent pulmonary resection for suspected primary lung cancer from 1997 to 2004. Of these, 76 patients had a history of HNC. The remaining 1557 patients were defined as controls. Categoric variables were analyzed with the chi2 test. Univariate and multivariate logistic regression analyses determined the variables related to aspiration pneumonia after pulmonary resection. RESULTS: Aspiration pneumonia occurred in 7 HNC patients (9.2%) versus 10 patients (0.6%) in the control group (p < 0.001). In the entire population with pulmonary resection, HNC history (p < 0.001; odds ratio (OR), 17.5; 95% confidence interval (CI), 6.0 to 50.6), and postoperative recurrent laryngeal nerve paralysis (p < 0.001; OR, 27.8; 95% CI, 5.2 to 148) were independent risk factors for aspiration pneumonia after pulmonary resection. Length of stay was longer in patients with aspiration pneumonia, with a median of 30 days (range, 10 to 258) versus 6 days (range, 0 to 374; p = 0.021). In the HNC patients, prior recurrent laryngeal nerve paralysis was predictive of aspiration pneumonia (p = 0.034; OR, 8.8; 95% CI, 1.1 to 65.4). CONCLUSIONS: Patients with HNC have an increased risk of aspiration pneumonia after pulmonary resection. Evaluation of swallowing function to identify aspiration is indicated in HNC patients before pulmonary resection to avoid the morbidity and prolonged hospitalization associated with aspiration pneumonia. PMID- 17126096 TI - Technical innovations of carinal resection for nonsmall-cell lung cancer. AB - BACKGROUND: We present our perioperative management of operable nonsmall-cell lung cancer invading the tracheobronchial bifurcation and the results obtained. METHODS: Fifty consecutive patients undergoing carinal surgery with radical lymphadenectomy over a 5-year period were studied. RESULTS: Eighteen patients (36%) were N2 and had chemoradiation (48 +/- 6 Gy) preoperatively. Surgery included 34 carinal pneumonectomies (24 right, 10 left), 11 carinal lobectomies (n = 6) or bilobectomies (n = 5), and 5 carinal resections, with (n = 3) and without (n = 2) reconstructions. Patients were ventilated through low tidal volume controlled techniques except during airway resection and reconstruction, during which the apneic (hyper) oxygenation techniques were used. High inspiratory oxygen concentrations, multiple collapse and reexpansions, hypoperfusion of the ipsilateral lung, and fluid overload were avoided. All patients but 1 were extubated in the operating room, 7 +/- 5 minutes after skin closure. Operative mortality (less than 30 days) and morbidity were 4% (n = 2) and 37% (n = 18), respectively. All resections but 1 (98%) R1 were complete. The number of resected nodes per patient was 9 +/- 2, and 7 (22%) of the 32 patients who had negative preoperative positron emission tomography results had micrometastatic mediastinal nodes. With a median follow-up of 38 months, actuarial 5-year and disease-free survivals were 51% and 47%, respectively. Disease-free survival was significantly affected by endobronchial extension (tracheobronchial angle invasion versus less than 0.5 cm from carina, p = 0.03) and nodal status (N0 versus N1-2, p = 0.02) in the multivariate analysis. CONCLUSIONS: Preoperative chemoradiation, carinal lobectomy, or left pneumonectomy, and radical lymphadenectomy do not worsen the therapeutic index of carinal surgery. The high incidence of micrometastatic nodes in positron emission tomography-negative patients justifies routine mediastinoscopy and radical lymphadenectomy. PMID- 17126097 TI - Differential pulmonary vein gases predict primary graft dysfunction. AB - BACKGROUND: Donor arterial blood gas measurements correlate poorly with lung allograft function in the recipient. We assessed the utility of reduced pulmonary vein gas (PVG) partial pressure of oxygen (PO2) in predicting the incidence of primary graft dysfunction. METHODS: While the donor was ventilated with 100% oxygen, superior and inferior pulmonary veins were directly aspirated bilaterally and pulmonary venous PO2 measured. A PO2 of less than 300 mm Hg was considered subnormal. These values were assessed for predictive value in terms of primary graft dysfunction in univariate and multivariate analysis. RESULTS: In 112 of the 201 lung and heart-lung transplants performed during the period January 2000 to December 2004, full PVGs were available for analysis. The number of pulmonary veins with sub-normal PVG correlated significantly with the incidence of severe primary graft dysfunction posttransplant in univariate (p = 0.01) and multivariate analysis (hazard ratio 2.35, p = 0.016). When analyzed separately, this correlation remained significant for recipients of single or bilateral transplants alone. No correlation existed between arterial PO2 at donor referral and incidence of primary graft dysfunction. Median duration of ventilation, intensive care unit stay, and 30-day and 90-day mortality were not significantly different for those with any subnormal PVG compared with those with all values in the normal range. CONCLUSIONS: Differential PVGs are a useful tool in the assessment of donor lung function before procurement. It is a helpful indicator of whether preischemic dysfunction is localized or diffuse, and can be used to predict the extent to which ischemia and reperfusion will exacerbate any existing abnormality. PMID- 17126098 TI - Invited commentary. PMID- 17126099 TI - Retrospective analysis of the clinical performance of anterior mediastinotomy. AB - BACKGROUND: Accurate staging of patients with lung cancer is imperative in generating an appropriate treatment strategy. This study examined the clinical performance of anterior mediastinotomy in staging patients with suspected left upper lobe non-small cell lung cancer. METHODS: This study was designed as a retrospective cohort. All patients with suspected left upper lobe cancer and otherwise normal computed tomography scan results were eligible. Patients with clinically unresectable disease (advanced disease or not fit for surgery) were excluded. After exclusions, 151 patients were stratified into two groups: 117 patients had cervical and anterior mediastinotomy as part of preoperative staging, and 34 had cervical mediastinoscopy only. The primary outcome was rate of preventable thoracotomy defined as thoracotomy during which either metastases to aortopulmonary or paraaortic lymph nodes, or mediastinal invasion was identified. RESULTS: The rate of preventable thoracotomy for the anterior mediastinotomy arm was 4 (3.4%) of 117, compared with 1 (2.9%) of 34 for cervical mediastinoscopy-only arm (p = 0.99). The rate of morbidity in the anterior mediastinotomy arm was 8 (6.8%) of 117, compared with 2 (5.8%) of 34 for the cervical mediastinoscopy-only arm (p = 0.99). Anterior mediastinotomy patients stayed in hospital 1 day longer (p = 0.008). Anterior mediastinotomy was successful at harvesting one or more lymph nodes in 67% of patients. Five patients (4.3%) who underwent anterior mediastinotomy were spared a thoracotomy by identification of metastases to aortopulmonary lymph nodes. CONCLUSIONS: In patients with suspected left upper lobe lung cancer and otherwise normal computed tomography scan results, anterior mediastinotomy does not significantly reduce the rate of preventable thoracotomy. PMID- 17126100 TI - Prognostic factors and surgical indications of pulmonary epithelioid hemangioendothelioma: a review of the literature. AB - BACKGROUND: Pulmonary epithelioid hemangioendothelioma is a rare vascular tumor of low-grade malignancy, the prognosis of which remains unpredictable. The purpose of this analysis based on 80 patients was to determine prognostic factors and to evaluate results of surgery. METHODS: We recorded data of 75 patients from reports published in the English and French literature using the terms "intravascular bronchoalveolar tumor" or "pulmonary epithelioid hemangioendothelioma" or a combination of both. We added to this database 5 more cases of pulmonary epithelioid hemangioendothelioma operated on in our thoracic surgery department from 1989 to 2005. Univariate and multivariate analyses of prognostic factors were performed using the log rank test and the Cox model. The factors we tested were age, sex, clinical symptoms, biologic and radiologic findings, and surgical treatment. RESULTS: There were 49 women and 31 men with a mean age of 39.7 years (range, 7 to 72 years). The 5-year survival probability was 60% (range, 47% to 71%). Univariate analysis showed that loss of weight, anemia, pulmonary symptoms, and more particularly pleural hemorrhagic effusions were significant factors of poor prognosis, with a median survival of less than 1 year. Multivariate analysis showed a statistically worse survival in patients with hemorrhagic symptoms (hemoptysis, p < 0.0001; pleural effusion, p < 0.0001). CONCLUSIONS: Pulmonary epithelioid hemangioendothelioma typically occurs among young patients. Surgery can be proposed in cases of unilateral single or multiple nodules. There is no single effective treatment in cases of bilateral multiple nodules. Lung transplantation should be evaluated in patients with vascular aggressivity with pleural hemorrhagic effusion and anemia. PMID- 17126101 TI - Surgical experience for the pulmonary artery sarcoma. AB - BACKGROUND: Pulmonary sarcoma arising from the pulmonary artery is a rare disease and its prognosis is disastrous. METHODS: Five patients who underwent surgery for pulmonary artery sarcoma were reviewed. RESULTS: All patients except one were initially diagnosed with pulmonary embolism. One patient with preoperative profound shock could not wean from cardiopulmonary bypass. Two patients are still surviving for 36 and 30 months, respectively. CONCLUSIONS: Early diagnosis and complete surgical resection is perhaps the best way to improve patients with pulmonary artery sarcoma. PMID- 17126102 TI - Toll-like receptor 4 mediates lung ischemia-reperfusion injury. AB - BACKGROUND: We have previously reported that nuclear factor (NF)-kappaB activation and inflammatory cytokine expression were involved in the development of lung ischemia-reperfusion injury (LIRI). Because Toll-like receptor 4 (TLR4) activates NF-kappaB-dependent transcription of inflammatory cytokine genes during myocardial ischemia-reperfusion injury, we examined whether absence of TLR4 in TLR4-deficient mice protects against LIRI. METHODS: Left lungs of wild-type (C57BL/6J) mice or TLR4-null (TLR4-/-) mice were made ischemic for 60 minutes and then reperfused for 180 minutes. Response to injury was quantified by tissue myeloperoxidase activity, vascular permeability ([125I]-bovine serum albumin extravasation), and leukocyte and inflammatory mediator accumulation in bronchoalveolar lavage expression. Lung homogenates were also analyzed for activation of mitogen-activated protein kinases and nuclear translocation of the transcription factors NF-kappaB and activator protein-1. RESULTS: After LIRI, lungs from TLR4-/- mice demonstrated a 52.4% reduction in vascular permeability (p = 0.001), a 52.6% reduction in lung myeloperoxidase activity (p = 0.006), and a marked reduction in bronchoalveolar lavage leukocyte accumulation when compared with lungs from wild-type mice. The TLR4-/- mice lungs, subjected to LIRI, also demonstrated marked reductions in amounts of several proinflammatory cytokines/chemokines in bronchoalveolar lavage samples. Phosphorylation of c-Jun NH2-terminal kinase, and activation of NF-kappaB and activator protein-1 were also significantly reduced in homogenates of lungs from TLR4-/- mice injured by ischemia and reperfusion (p < 0.05). CONCLUSIONS: These data suggest that TLR4 plays a role in LIRI. Thus, TLR4 may be a potential therapeutic target to minimize ischemic-reperfusion-induced tissue damage and organ dysfunction. PMID- 17126103 TI - Invited commentary. PMID- 17126104 TI - Antegrade versus retrograde isolated lung perfusion: doxorubicin uptake and distribution in a sarcoma model. AB - BACKGROUND: Antegrade and retrograde doxorubicin-based isolated lung perfusions were compared in rodents bearing a sarcomatous tumor in the perfused lung. Inasmuch as these tumors derive their vascularization from the bronchial artery system, we hypothesized that retrograde isolated lung perfusion through the pulmonary vein might result in an improved tumor drug uptake. METHODS: Single pass antegrade (n = 9) and retrograde (n = 9) isolated left lung perfusions were performed with 100 microg of doxorubicin in Fischer rats 10 days after subpleural tumor cell injection. The perfusion, washout, and recirculation times were 20, 10, and 60 minutes, respectively, followed by harvesting of the lung. The doxorubicin concentration and compartmental distribution in the tumor and in normal parenchyma of each perfused lung were measured by high-pressure liquid chromatography (6 animals of each group) and fluorescence microscopy (3 animals of each group). RESULTS: Doxorubicin concentration and pattern of doxorubicin based fluorescence signaling were comparable for both perfusion techniques in normal lung tissue. Antegrade and retrograde isolated lung perfusion resulted in similar tumor drug uptake, which was lower than in normal lung parenchyma, and in weak and sporadic fluorescence signaling emerging from the tumor periphery and from blood vessels situated within the tumor tissue. CONCLUSIONS: Retrograde isolated lung perfusion did not confer a better doxorubicin uptake in the tumor as compared with antegrade lung perfusion despite the fact that the tumor vascularization in this model is based on the bronchial artery circulation. PMID- 17126105 TI - Invited commentary. PMID- 17126106 TI - Mortality increases for octogenarians undergoing esophagogastrectomy for esophageal cancer. AB - BACKGROUND: As the general population ages, it becomes increasingly important to understand the potential contribution of chronologic age to mortality after esophagectomy. Because this risk is poorly defined, we sought to determine whether extreme age (>80 years) is an independent risk factor after esophagectomy. METHODS: We analyzed a prospectively maintained, single institution database of 858 consecutive patients who underwent esophagectomy between January 1996 and May 2005. Data evaluated included patient demographics, medical comorbidity, types of resections performed, length of stay, postoperative adverse events, and overall survival. We used univariate, multivariate, and Kaplan-Meier analysis to determine the influence of age on postoperative morbidity, in-hospital survival, and overall survival. RESULTS: Of 858 patients, 31 (10 female, 21 male) were older than 80 years of age. Preliminary analysis indicated that patients younger than 50 years (n = 107) had significantly fewer comorbidities; these were excluded from the analysis. In the remaining 751 patients, the age older than 80 cohort was compared with patients aged 50 to 79. Patients aged 50 to 79 were grouped because of similar characteristics (length of stay, hospital death). There were no significant differences in comorbidities, types of resections, or postoperative complication type or severity between the two groups. Postoperative death, length of stay, and survival, however, were significantly worse in patients older than 80. In a logistic regression model controlling for comorbidity, age older than 80 was significantly associated with increased perioperative mortality (hazard-ratio, 3.9; p < 0.01). CONCLUSIONS: Patients older than 80 years have increased mortality risk after esophagectomy, independent of comorbidity. Octogenarian status should be a consideration in the management of these patients. PMID- 17126107 TI - Feasibility and outcomes of an early extubation policy after esophagectomy. AB - BACKGROUND: Although early extubation of esophagectomy patients has been found to be feasible, safe, and associated with low morbidity, there is no uniform standard of care among high volume centers. Our objective is to examine a contemporary series of esophagectomies and identify the feasibility and outcome of an early extubation policy. METHODS: This study is a retrospective review of all patients who underwent esophagectomy between January 2003 and December 2004 at the Massachusetts General Hospital. One hundred and two patients were analyzed from 129 consecutive patients who underwent esophagectomy and subsequently divided in two groups: The early extubation group was extubated in the operating room and the late extubation group was extubated in the intensive care unit (ICU). RESULTS: Ninety percent were extubated early. Although most patients underwent a transthoracic or thoracoabdominal esophagectomy, the operative approach did not influence failure to extubate. Neoadjuvant therapy was not predictive of extubation failure. Most patients age 70 or greater (86%) were extubated early. There were three nonelective reintubations in the early extubation group secondary to acute respiratory distress syndrome. The median length of stay was 11 days and median ICU stay was one day. The 30-day mortality was 1.9% and the median survival was 28 months. CONCLUSIONS: Attention to restricted intraoperative fluid balance, limited blood loss, anesthetic technique, and epidural use permit most patients undergoing esophageal resection to be safely extubated immediately postresection in the operating room. PMID- 17126108 TI - Brain metastases from esophageal cancer: a phenomenon of adjuvant therapy? AB - BACKGROUND: Brain metastases from esophageal cancers are uncommon, yet our impression was that they occurred more frequently than expected after esophagectomy plus adjuvant therapy. Therefore, we determined (1) incidence and prevalence of, risk factors for, and survival after development of brain metastases following esophagectomy for esophageal cancer, and (2) their association with adjuvant therapy. METHODS: From 1985 to 2002, 403 patients (52%) underwent esophagectomy alone and 369 esophagectomy plus adjuvant therapy (118 [15%] preoperative only, 124 [16%] postoperative only, and 127 [16%] both). Hazard-function methodology was used to characterize time-related occurrence of brain metastases and risk factors. Inferences were confirmed by propensity analysis. RESULTS: Twenty-nine patients developed brain metastases, 20 within 1 year; 6 had undergone surgery alone, and 23 had adjuvant therapy. Prevalence was 2.5% 5 years after surgery alone, but 8.4%, 7.0%, and 18.4% after preoperative adjuvant therapy only, postoperative adjuvant therapy only, and both, respectively (p < 0.0001). Greater number of locoregional lymph node metastases was associated with brain metastases after surgery alone (p = 0.04). Distant metastases (p = 0.03) and both preoperative and postoperative adjuvant therapy (p = 0.004) were risk factors. Median survival after diagnosis of brain metastases was 3.5 months. Postesophagectomy propensity-matched survival was shorter after adjuvant therapy than after surgery alone; thus, time available for developing brain metastases after surgery alone was slightly lower. CONCLUSIONS: A dose related increased incidence of brain metastases after adjuvant therapy for esophageal cancer cannot be explained by increased longevity. Adjuvant therapy itself, not just advanced disease, appears to create conditions conducive to developing these rapidly fatal metastases. PMID- 17126109 TI - Reinforcement of esophageal anastomoses with an extracellular matrix scaffold in a canine model. AB - BACKGROUND: The gastric pull-up procedure, a standard intervention after radical esophagectomy, is associated with high morbidity and mortality due to leaks and stricture. A previous preclinical study showed that an extracellular matrix (ECM) scaffold with autologous muscle tissue could be used to repair a complete circumferential defect in the cervical esophagus. The aim of the present study was to determine if healing of end-to-end anastomoses of the esophagus could be improved by reinforcement with an ECM scaffold. METHODS: Twelve female mongrel dogs underwent a complete transection of either the cervical esophagus (n = 6) or the gastroesophageal junction (n = 6). A portion of the endomucosa at the anastomotic site was resected and replaced with an ECM scaffold in contact with the subjacent muscle and the muscle was anastomosed. The measured end points included macroscopic and microscopic evaluation and quantification of the esophageal diameter at the anastomotic site. RESULTS: No anastomotic leaks or systemic complications were observed in the ECM-treated animals. Morphologic findings in both groups showed complete mucosal covering of the surgery site. The remodeled esophageal tissue showed angiogenesis and complete epithelialization. Intact, organized layers of muscle tissue were present between the native muscularis externa and the submucosal layer and effectively bridged the transected ends. CONCLUSIONS: The ECM scaffold altered the default mechanism of esophageal repair. Scar tissue formation with associated stricture was virtually eliminated, and the esophageal healing response was characterized by the replacement with structurally normal tissue layers. These findings suggest that the high morbidity rate associated with esophagectomy procedures may be reduced by this ECM augmentation procedure at the anastomotic site. PMID- 17126110 TI - Determinants of left ventricular dysfunction after anatomic repair of congenitally corrected transposition of the great arteries. AB - BACKGROUND: Early results for anatomic repair of congenitally corrected transposition of the great arteries are excellent with respect to right ventricular and tricuspid valve function. However, development of left ventricular (systemic ventricle) dysfunction late after repair remains a concern. In this study we sought to determine factors leading to late impairment in left ventricular performance. METHODS: From August 1992 to July 2005, 44 patients (median age at surgery, 1.6 years; range, 0.6 to 39.6 years) with congenitally corrected transposition of the great arteries had anatomic repair. Left ventricular function and mitral regurgitation were evaluated by echocardiography at follow-up. Twenty-three patients had a Rastelli procedure, and 21 underwent an arterial switch. Twelve patients (27%) were pacemaker dependent at latest follow up. RESULTS: Early mortality was 4.5% (n = 2) with 1 late death as a result of leukemia. Median follow-up was 3.0 years (range, 7 days to 12.4 years). Left ventricular function remained unchanged (normal) in 35 patients, improved in 1 patient, and deteriorated in 8 patients (18%). Mitral regurgitation was unchanged in 30 patients, improved in 6 patients, and worsened in 8 patients (18%). Development of left ventricular dysfunction was significantly associated with pacemaker implantation (p = 0.005) and a widened QRS (>20% > 98% percentile of normal; p = 0.03). CONCLUSIONS: Anatomic repair of congenitally corrected transposition can be performed with low operative mortality. However, late left ventricular dysfunction is not uncommon, with higher incidence in those requiring pacing and with a prolonged QRS. Resynchronization may be of value in patients requiring a pacemaker. PMID- 17126111 TI - Revascularization of left anterior descending artery with drug-eluting stents: comparison with minimally invasive direct coronary artery bypass surgery. AB - BACKGROUND: The proximal left anterior descending artery (LAD) is a challenging area for percutaneous interventions; therefore, coronary artery bypass grafting is often considered and sometimes performed even in patients with single-vessel disease involving the proximal LAD. This study compares mid-term results of LAD revascularization using drug-eluting stents (Cypher) with minimally invasive direct coronary artery bypass grafting (MIDCAB). METHODS: From May 2002 to December 2003, 376 consecutive patients underwent myocardial revascularization of the LAD, 272 by Cypher and 104 by MIDCAB. After matching for age, sex, and extent of coronary artery disease, two groups of 83 patients each were used to compare the two revascularization modalities. The groups were similar; however, ejection fraction of less than 0.35 was more prevalent in the MIDCAB group and prior percutaneous coronary intervention in the Cypher group. RESULTS: Thirty-day mortality was 1.1% in the MIDCAB and 0% in the Cypher group. Mean follow-up was 22.5 months. Two late cardiac deaths occurred in the MIDCAB group and one in the Cypher group (p = NS). Angina returned in 35% of the Cypher group and in 8.4% of the MIDCAB group (p < 0.001). There were 14 (16.8%) reinterventions in the Cypher compared with three (3.6%) in the surgical group (p = 0.005). Cox proportional hazard model showed that assignment to the Cypher group was the only independent predictor of reangina (hazard ratio [HR], 6.17, 95% confidence interval [CI], 2.46 to 15.4). Treatment with Cypher was also an independent predictor of reintervention (HR 8.26, 95% CI, 1.68 to 40). CONCLUSIONS: Despite improved results of percutaneous interventions with Cypher to the LAD, mid-term clinical outcome of patients treated with MIDCAB was better. PMID- 17126112 TI - Relation of the Sequential Organ Failure Assessment score to morbidity and mortality after cardiac surgery. AB - BACKGROUND: Organ dysfunction evaluation using Sequential Organ Failure Assessment (SOFA) has been shown to predict mortality and morbidity in adult cardiac surgical patients with prolonged recovery. The purpose of this study was to evaluate the utility of SOFA in prediction of mortality and morbidity in a cohort of heterogeneous consecutive adult cardiac surgical patients. METHODS: A prospective study of 857 consecutive patients entering in a single cardiac postoperative intensive care unit was assigned during the year 2004. The European System for Cardiac Operative Risk Evaluation (EuroSCORE) of each patient was assessed preoperatively. SOFA was calculated daily until intensive care unit discharge or for a maximum of 7 days. SOFA change between the first and the third postoperative day, maximum SOFA during the first 3 days, and maximal SOFA were calculated. Length of intensive care unit stay and 30-day mortality were assessed. RESULTS: Maximum SOFA during the first 3 days and maximal SOFA predicted 30-day mortality (area under the curve, 0.763 and 0.779, respectively) also correlated with the length of intensive care unit stay (p < 0.001 and p < 0.001, respectively). The EuroSCORE predicted both mortality and intensive care unit stay (p < 0.0001 and p < 0.0001). The correlation coefficient between the EuroSCORE and maximum SOFA during the first 3 days or maximal SOFA was low (r = 0.34 and 0.33, respectively, p < 0.0001 and p = 0.0001). CONCLUSIONS: The SOFA score is an independent predictor of mortality and length of stay in cardiac surgical patients. The SOFA score is associated with mortality and morbidity even when assessed in the early postoperative period after adult cardiac surgery. PMID- 17126113 TI - Invited commentary. PMID- 17126114 TI - Long-term outcome of survivors of prolonged intensive care treatment after cardiac surgery. AB - BACKGROUND: The relative impact of perioperative risk profile and postoperative complications on long-term outcome in cardiac surgical patients is currently unclear. The aim of this work was to assess the relative predictive value of the European System for Cardiac Operative Risk Evaluation (EuroSCORE) and Sequential Organ Failure Assessment (SOFA) on long-term event-free survival in this patient population. METHODS: Preoperative and postoperative variables, EuroSCORE and SOFA, 30-day mortality, and long-term mortality or hospital admission for cardiovascular events were assessed in 115 consecutive cardiac surgical patients in whom multiorgan dysfunction syndrome developed postoperatively. RESULTS: Mean age was 70 +/- 8 years, 41% were women, EuroSCORE averaged 7.87 +/- 3.99, and postoperative stay in the intensive care unit was 10.3 +/- 8.2 days. In-hospital 30-day mortality was 10.4% (n = 12). During 1998 person-months follow-up, 12 (11.6%) of 103 patients discharged alive died, and 46 (44.7%) met the combined end point of all-cause death or cardiovascular admission. By Cox multivariate analysis, maximum SOFA (hazard ratio [HR], 2.17; 95% confidence interval [CI], 1.34 to 3.51) and maximum cardiovascular score (HR, 2.35; 95% CI, 1.22 to 4.51) independently predicted all-cause mortality. EuroSCORE (HR, 1.33; 95% CI, 1.01 to 1.76), maximum cardiovascular score (HR 2.09; 95% CI 1.41 to 3.10), and maximum liver score (HR 2.67; 95% CI, 1.46 to 4.86) were independently associated with the combined end point. CONCLUSIONS: High-risk cardiac surgical patients with postoperative multiorgan dysfunction syndrome show excess mortality and cardiovascular morbidity after hospital discharge. Combined preoperative and postoperative risk stratification identifies patients with the highest likelihood of death or early readmission. PMID- 17126115 TI - Invited commentary. PMID- 17126116 TI - Donor tricuspid annuloplasty during orthotopic heart transplantation: long-term results of a prospective controlled study. AB - BACKGROUND: Development of tricuspid regurgitation after orthotopic heart transplantation can cause heart failure along with renal and hepatic impairment and portends a poor prognosis. If tricuspid regurgitation causes significant symptoms, tricuspid valve repair or replacement is often required. This study was designed to study the effects of prophylactic tricuspid valve annuloplasty (TVA) during orthotopic heart transplantation on long-term survival, renal function, and amount of tricuspid regurgitation. METHODS: Between April 1997 and March 1998, 60 patients (aged 18 to 70 years; 22 female) randomly received either standard bicaval orthotopic heart transplantation (group STD; n = 30) or bicaval orthotopic heart transplantation with DeVega TVA (group TVA; n = 30). Tricuspid valve annuloplasty was performed on the donor heart before implantation using pledgeted 2-0 polypropylene and sized to an annulus of 29 mm. Echocardiographic measurements, laboratory values, and hemodynamics were obtained prospectively and reviewed by an independent data analyst. RESULTS: Follow-up of patients as of December 2003 was complete. Although there was a perioperative mortality advantage in group TVA, there was no difference between groups in long-term survival. At the end of the study, however, there was a statistical difference (group STD versus group TVA, p < 0.05) with regard to cardiac mortality (7 of 30 versus 3 of 30), average amount of tricuspid regurgitation (1.5 +/- 1.3 versus 0.5 +/- 0.4), percentage of patients with 2+ or greater tricuspid regurgitation (34% versus 0%), serum creatinine (2.9 +/- 2.0 versus 1.8 +/- 0.7), and difference in serum creatinine over baseline (2.0 +/- 2.1 versus 0.7 +/- 0.8). CONCLUSIONS: Prophylactic DeVega TVA of the donor heart is durable and decreases the incidence of cardiac-related mortality and tricuspid regurgitation after orthotopic heart transplantation. In addition, there is improved protection of renal function. Considering the ease and safety of TVA and its advantages, it should be performed as a routine adjunct to orthotopic heart transplantation. PMID- 17126117 TI - Large annuloplasty rings facilitate mitral valve repair in Barlow's disease. AB - BACKGROUND: Excess leaflet tissue in Barlow's disease predisposes patients to left ventricular outflow tract obstruction and residual mitral regurgitation after mitral valve repair as a result of systolic anterior motion of the anterior mitral leaflet. In addition to conventional repair techniques such as sliding plasty and leaflet shortening, our approach in Barlow's disease has included the use of large remodeling annuloplasty rings (up to size 40 mm). We report our experience with the use of large rings in Barlow's disease. METHODS: From January 2002 to December 2005, 67 patients with Barlow's disease (46 men and 21 women; median age, 55 years; range, 22 to 85 years), mean ejection fraction 0.55 +/- 0.08, and grade 3+ or greater mitral regurgitation underwent mitral valve repair. All had Carpentier type II leaflet dysfunction, with anterior (n = 2), posterior (n = 41), or bileaflet (n = 24) prolapse. Predominant reconstructive techniques were posterior leaflet sliding plasty/plication (n = 65), anterior leaflet triangular resection (n = 16), and chordal transfer (n = 25). Concomitant procedures included coronary artery bypass grafting surgery (n = 8), tricuspid valve repair (n = 20), aortic valve replacement (n = 3), and CryoMaze (n = 22). RESULTS: Mitral valve repair was successfully completed in all patients. Annuloplasty ring size was 36 mm (n = 17), 38 mm (n = 22), and 40 mm (n = 28). Predischarge transthoracic echocardiography showed absence of systolic anterior motion (n = 67), no or trace mitral regurgitation (n = 62), and mild mitral regurgitation (n = 5). There was no operative mortality. CONCLUSIONS: Mitral valve repair can be predictably performed in Barlow's disease with excellent early outcomes. Large annuloplasty rings help minimize the risk of systolic anterior motion and are an important adjunct to established repair techniques in this patient cohort with large annular size and excess leaflet tissue. PMID- 17126118 TI - Preoperative and late postoperative mitral regurgitation in ventricular reconstruction: role of local left ventricular deformation. AB - BACKGROUND: We sought to analyze the characteristics of local left ventricular deformation related to functional mitral regurgitation (MR) in post-anterior myocardial infarction scar, and to evaluate how local remodeling contributes to late development of MR after surgical ventricular reconstruction by endoventricular circular patch plasty repair. METHODS: Two hundred twenty-one consecutive patients (aged 60 +/- 9 years; 193 males) with previous transmural anterior infarction underwent heart catheterization both before and 1 year after endoventricular circular patch plasty repair. Preoperative global left ventricular shape determinants (eccentricity and circularity indexes), regional curvature and wall motion (centerline), and both preoperative and 1-year postoperative hemodynamic parameters (volumes, ejection fraction, capillary wedge and pulmonary artery pressures) were calculated. RESULTS: Forty-eight patients had (MR patients), and 173 did not have (NoMR patients) angiographic MR grade 2 or more preoperatively; at follow-up, 30 NoMR patients had MR (late MR [LMR]). Before surgery, MR patients had larger left ventricular volumes, higher capillary wedge and mean pulmonary artery pressures, and lower ejection fraction and cardiac index. The LMR patients had similarly high capillary wedge and pulmonary artery pressures as MR patients; otherwise, they did not differ from NoMR patients. Mitral regurgitation patients had wider lateral wall akinesia and greater inferior wall asynergy; the inferobasal region was hypokinetic in LMR patients. In MR patients, inferior wall systolic curvature was less negative; the inferobasal region had a more positive curvature in LMR patients. CONCLUSIONS: Local deformation of the inferior wall with loss of systolic inward bending is associated with functional MR, while asynergy and systolic deformation of the inferobasal region and high capillary wedge pressure are prognostic signs of MR development late after endoventricular circular patch plasty repair. PMID- 17126119 TI - Invited commentary. PMID- 17126120 TI - Clinical profile and natural history of 453 nonsurgically managed patients with severe aortic stenosis. AB - BACKGROUND: Severe aortic stenosis (AS) is a surgically correctable condition. However, aortic valve replacement (AVR) is not offered to many patients with severe AS for various reasons. We investigated the profile and survival patterns of patients with severe AS who did not have AVR. METHODS: Our echocardiographic database was screened for patients with severe AS, defined as a Doppler estimated aortic valve area of 0.8 cm2 or less between 1993 and 2003. Seven hundred and forty patients with severe AS were identified, of whom 453 patients had no AVR through the follow-up period, forming the study cohort. These patients were comprehensively characterized by obtaining clinical, pharmacologic, and surgical data through a comprehensive chart review and extracting survival data from the National Death Index. RESULTS: Patient characteristics were as follows: age 75 +/ 13 years, 48% male, left ventricular (LV) ejection fraction 52 +/- 21%, coronary artery disease in 34%, hypertension in 35%, serum creatinine level greater than 2 mg/dL in 11%, and diabetes mellitus in 14%. The survival at 1 year, 5 years, and 10 years was 62%, 32%, and 18%, respectively. The univariate predictors of reduced survival were advanced age, low LV ejection fraction, heart failure, elevated serum creatinine level, severe mitral regurgitation, and pulmonary hypertension; and the independent predictors were advanced age, low LV ejection fraction, heart failure, elevated serum creatinine level, and systemic hypertension. Concomitant pharmacotherapy had no impact on survival. CONCLUSIONS: Conservatively treated patients with severe AS have a grave prognosis, and it is worse in the presence of advanced age, LV dysfunction, heart failure, and renal failure. PMID- 17126121 TI - Invited commentary. PMID- 17126122 TI - Malignant natural history of asymptomatic severe aortic stenosis: benefit of aortic valve replacement. AB - BACKGROUND: Patients with asymptomatic severe aortic stenosis (AS) are reported to have a benign prognosis and hence the American College of Cardiology/American Heart Association guidelines do not recommend aortic valve replacement (AVR) for patients with isolated asymptomatic severe AS. However, symptoms are subjective and would depend upon patient's life style. We examined the natural and unnatural history of initially asymptomatic patients with severe AS. METHODS: A search of our echocardiographic database between 1993 and 2003 yielded 740 patients with severe AS defined as aortic valve area 0.8 cm2 or less. Thorough chart reviews were conducted to collect clinical and pharmacologic data. Of these, 338 patients were asymptomatic at the initial encounter forming the study cohort. RESULTS: Patient characteristics were the following: age 71 +/- 15 years, males 51%, aortic valve area 0.72 +/- 0.17 cm2, left ventricular ejection fraction 0.59 +/- 0.17. Ninety-nine (29%) patients had AVR during a mean follow-up of 3.5 years. Survival at 1, 2, and 5 years in the nonoperated patients were 67%, 56%, and 38%, respectively, compared with 94%, 93%, and 90% in those who underwent AVR (p < 0.0001). The Cox regression model was used to adjust for the effect of 18 clinical, echocardiographic, and pharmacologic variables on survival. The adjusted hazard ratio for death with AVR was 0.17 (95% confidence interval [CI] 0.10 to 0.29). In the nonoperated group, renal insufficiency (risk ratio [RR] 3.1, 95% CI 1.5 to 6.6), beta blocker use (RR 0.52, 95% CI 0.31 to 0.88), statin use (RR 0.52, 95% CI 0.27 to 0.99), age (per year RR 1.03, 95% CI 1.02 to 1.05), and left ventricular ejection fraction (per % RR 0.99, 95% CI 0.98 to 1.00) were found to be the independent predictors of mortality. The benefit of AVR was further supported by sensitivity and propensity score analyses. CONCLUSIONS: Our observational data indicate that the natural history of asymptomatic AS is not benign and that survival is dramatically improved by AVR. Survival of the asymptomatic unoperated or nonoperable patients may potentially be improved by the use of beta blockers and statins. PMID- 17126123 TI - What prosthesis should be used at valve re-replacement after structural valve deterioration of a bioprosthesis? AB - BACKGROUND: The fate of bioprostheses (BP) and mechanical prostheses (MP) after valve re-replacement for bioprostheses is not well-documented. This research compares the late fate of these two valve types after valve re-replacement for structural valve deterioration (SVD) of a bioprosthesis. METHODS: Between 1975 and 2000, 298 patients had successful aortic valve re-replacements (AVRR) (BP n = 149, average age = 67.1 +/- 12.3 years; MP 149, 58.9 +/- 10.9) and 442 patients had successful mitral valve re-replacements (MVRR) (BP 155, 65.8 +/- 14.1; MP 287, 60.8 +/- 11.7) after SVD of a previous BP. Follow-up was five years in all groups. RESULTS: (1) Aortic position (AVRR): Survival favored MP over BP overall, at 10 years (70.3 +/- 5.4% vs 56.7 +/- 5.7%, p = 0.0220). This survival advantage was seen to be significant only in patients less than 60 years of age (at 10 years, 85.3 +/- 4.9% vs 59.2 +/- 9.8%, p = 0.038). No significant difference in survival between the two valve types was observed in patient age groups greater than 60 years of age. Freedoms from valve-specific complications, including reoperation for SVD-thrombosis, major thromboembolism and hemorrhage, and valve related mortality were not significantly different between the two groups overall. (2) Mitral position (MVRR): Survival favored MP over BP overall (58.6 +/ 4.2% vs 42.1 +/- 5.2%, p = 0.0011), and in patients greater than 70 years of age (32.8 +/- 8.9% vs 16.7 +/- 7.1%, p = 0.008). Freedoms from valve-specific complications and valve-related mortality favored MP over BP. CONCLUSIONS: There was no clinical performance difference between mechanical and bioprosthetic valves in patients greater than 60 years of age upon AVRR. Mechanical valves generally outperformed bioprosthetic valves in all age groups in MVRR. PMID- 17126124 TI - Results of aortic valve replacement with a new supra-annular pericardial stented bioprosthesis. AB - BACKGROUND: The stented Soprano valve (Sorin Biomedica S.pA., Saluggia, Italy) is a pericardial tissue valve designed for a totally supra-annular implant with the aim of improving hemodynamic performance, particularly in patients with a reduced aortic annulus. However, its hemodynamics and early clinical outcome are poorly understood. METHODS: Seventy-seven patients (mean age 76 +/- 5 years, 56% males) underwent aortic valve replacement with the Soprano valve. All patients were monitored with clinical examination and serial echocardiography at 1, 6, and 12 months and yearly afterward. RESULTS: At preoperative echocardiography, average left ventricular outflow tract diameter was 2.1 +/- 0.2 cm. At operation, 35% of patients received a 20-mm valve, 54% a 22-mm valve, and 11% a 24-mm valve. At 6 month follow-up, peak and mean transprosthetic gradients were 18 +/- 8 and 9 +/- 4 mm Hg, respectively; effective orifice area (EOA) and EOA index were 1.84 +/- 0.6 cm2 and 0.9 +/- 0.2 cm2/m2, respectively. Incidence of patient-prosthesis mismatch (ie, EOA index < 0.85 cm2/m2) was 23%, with no case of severe mismatch (ie, EOA index < 0.6 cm2/m2). In addition, left ventricular hypertrophy showed a significant regression (mass index from 214 +/- 98 g to 129 +/- 41 g; p = 0.001), and ejection fraction increased (from 58% +/- 17% to 67% +/- 8%; p = 0.001). Cumulative follow-up was 7.9 months per patient. Thirty-day mortality rate was 2.6% (2 of 77). Cumulative survival at 6, 12, and 24 months was 92% +/- 3.7%, 85% +/- 5.7%, and 85% +/- 5.7%, respectively. Two patients experienced embolic episodes. One case of anticoagulant-related bleeding occurred. CONCLUSIONS: In our series, the Soprano bioprosthesis showed a good hemodynamic performance, low incidence of patient-prosthesis mismatch, and favorable early clinical results. PMID- 17126125 TI - Venovenous extracorporeal life support after pulmonary endarterectomy: indications, techniques, and outcomes. AB - BACKGROUND: Pulmonary endarterectomy is the accepted therapy for thromboembolic pulmonary hypertension. A recognized complication of this surgery is the postoperative development of reperfusion edema, a potentially fatal cause of respiratory failure. Because reperfusion edema can be a reversible process, temporizing support measures may be life saving. METHODS: We retrospectively reviewed our experience with venovenous extracorporeal life support (V-V ECLS) from July 1990 to February 2006, in 20 adult patients (mean age 50.5 +/- 14.5 years) presenting with potentially reversible respiratory failure after pulmonary endarterectomy. This subset of patients comprised 1.12% of our total pulmonary endarterectomy experience during that time (1,790 cases). RESULTS: Overall in hospital survival was 30.0% for patients requiring ECLS support after pulmonary endarterectomy versus 94.2% for patients who underwent pulmonary endarterectomy alone during the same timeframe. V-V ECLS was instituted at a mean of 86.8 hours after surgery. The mean duration of V-V ECLS was 123.4 +/- 71.3 hours. The most common cause of death in ECLS patients after pulmonary endarterectomy was pulmonary hemorrhage. Survival was greater in patients cannulated within 120 hours of surgery (46.2% survival; 6 of 13 patients) compared with those cannulated after 120 hours (0 of 7 patients). Multiple logistic regression identified long duration of mechanical ventilation pre-ECLS and severity of preoperative pulmonary hypertension together as predictors of mortality. CONCLUSIONS: A small subset of patients undergoing pulmonary endarterectomy develop temporary life-threatening respiratory failure secondary to severe reperfusion edema. In those patients with satisfactory hemodynamic outcome, V-V ECLS is a therapeutic option when all other conventional strategies have been exhausted. PMID- 17126126 TI - Invited commentary. PMID- 17126127 TI - Long-term results from a 12-year experience with endovascular therapy for thoracic aortic disease. AB - BACKGROUND: Endovascular approaches promise to revolutionize therapy for thoracic aortic disease. This study describes a long-term analysis of endovascular thoracic aortic repair. METHODS: Seventy-three patients (mean age, 67.4 years) underwent endovascular thoracic aortic repair from 1993 to 2005. Indications for intervention included aneurysm (38%), dissection (23%), or penetrating ulcer or pseudoaneurysm (34%). Rupture was present in 16 patients (22%). Seventy-one percent were considered high risk for open surgery for reasons of age or comorbid conditions. Treated segments included ascending aorta (n = 1), distal arch (n = 24), and proximal (n = 50) or distal (n = 55) descending aorta. The total descending thoracic aorta was covered in 31 patients. Procedural success was achieved in 96%. Devices were delivered by femoral (79%), retroperitoneal iliac (18%), or carotid (2.7%) exposure. Devices used included Excluder (n = 30), Talent (n = 23), Zenith (n = 3), AneuRx (n = 5), and custom-fabricated (n = 14). Follow-up was 100% complete. RESULTS: Thirty-day mortality was 5.5%. Significant morbidity included stroke (8.2%) and need for dialysis (4.1%). Although 3 patients had transient spinal cord ischemia (4.1%), none had permanent sequelae. Intervention for fusiform aneurysm was independently associated with a composite end point of 30-day mortality, need for dialysis, and stroke (p = 0.015). Eight patients (11%) had new or persistent endoleaks, and aortic reintervention was performed in 7 patients (9.6%). Mean survival for the entire cohort was 46.8 +/- 5.1 months. Intervention for penetrating ulcer or pseudoaneurysm (p = 0.045) was independently associated with long-term all-cause mortality. CONCLUSIONS: An endovascular approach produces acceptable results for a broad range of thoracic aortic disease. However, the potential for endoleak or need for reintervention mandates continued close follow-up to achieve satisfactory long-term results. PMID- 17126128 TI - Axillary cannulation for proximal aortic surgery is as safe in the emergent setting as in elective cases. AB - BACKGROUND: Right axillary artery cannulation and selective antegrade cerebral perfusion (SCP) have become well-described strategies in the surgical treatment of proximal aortic disease. Many series report increases in adverse outcomes with SCP used in emergent settings. We compare outcomes in elective and emergent patients. METHODS: Over 21 months, SCP through right axillary cannulation with a side graft was performed in 61 patients. Thirty-three percent (20 of 61) underwent emergent operation for Stanford type A dissection or intramural hematoma, including 3 of 20 (4.7%) with pericardial tamponade; the remainder of SCP (41 of 61) was elective. The mean follow-up was 9.1 +/- 0.40 months. RESULTS: Selective antegrade cerebral perfusion was used in 20 of 22 emergent cases (91%), with 2 unsuccessful cannulation attempts, and no peripheral arterial dissections encountered. The SCP flows averaged 16.3 +/- 0.71 cc x kg(-1) x min(-1) for a mean perfusion period of 26.1 +/- 1.9 minutes. The average cardiopulmonary bypass time for all patients was 173 +/- 11 minutes. Average hospital stay was 8.1 +/- 0.80 days. One case (1.3%) of permanent and 3 cases (4.8%) of temporary neurologic dysfunction occurred in SCP patients. The hospital mortality rate for emergent SCP cases (2 of 20, 10%) was not statistically different from the mortality rate for elective SCP cases (3 of 41, 7.3%, p = not significant), with no difference in complication rates. All 3 SCP patients with preoperative tamponade survived without complication. Cerebral oximetry data showed a trend toward decreased left-sided (contralateral) scalp perfusion. There was no association of emergent status with neurologic dysfunction, death, or any other adverse outcome. CONCLUSIONS: Axillary cannulation and SCP in the surgical treatment of proximal aortic pathology is safe in both elective and emergent settings. PMID- 17126129 TI - Clinical indication for use and outcomes after inhaled nitric oxide therapy. AB - BACKGROUND: Inhaled nitric oxide (iNO) use is widespread, but the long-term outcomes after therapy in adult patients remain unknown. METHODS: All 376 patients receiving perioperative iNO (excluding pediatric and interventional cardiology procedures) at Columbia University Medical Center were prospectively followed from 2000 to 2003. Survival data were collected from chart review. RESULTS: Inhaled nitric oxide was used to treat pulmonary and right ventricular failure in patients undergoing orthotopic heart transplantation (OHT, n = 67), orthotopic lung transplantation (n = 45), cardiac surgery (n = 105), and ventricular assist device placement (n = 66), and for hypoxemia in other surgery (n = 34) and medical patients (n = 59). Average follow-up was 2.9 +/- 1.0 years. Overall mortality was lowest when iNO was used after OHT (25.4%) and orthotopic lung transplantation (37.8%), intermediately after cardiac surgery (61%), ventricular assist device (62%), and other surgery patients (75%), and highest among medical patients (90%; all p < 0.005). The cost of iNO therapy was lower in transplantation versus medical patients, with a trend toward shorter duration of use. In multivariate analysis, respiratory failure and use in non-OHT were independent predictors of mortality (both p = 0.001). A risk score greater than 1 (score = non-OHT use 1, plus right ventricular failure 1) predicted a mortality of 76.5% versus 37.2% (p < 0.001). CONCLUSIONS: Use of iNO for pulmonary hypertension in patients undergoing OHT and orthotopic lung transplantation was associated with a significantly lower overall mortality rate compared with its use after cardiac surgery or for hypoxemia in medical patients. Inhaled nitric oxide does not appear to be cost effective when treating hypoxemia in medical patients with high-risk scores and irreversible disease. PMID- 17126130 TI - Preoperative C-reactive protein predicts mid-term outcome after cardiac surgery. AB - BACKGROUND: C-reactive protein (CRP) is a known risk factor for cardiovascular events in the healthy population and in patients with coronary artery disease. High CRP levels before cardiac surgery are associated with worse short-term outcome, but its role after discharge home remains unknown. The study objective was to evaluate the effect of CRP on short-term and mid-term outcome after cardiac surgery. METHODS: From August 2000 to May 2004, values for preoperative CRP were available for 597 unselected patients undergoing cardiac operations. CRP was used to divide this cohort in two groups: a low inflammatory status (LHS) group of 354 patients with CRP of less than 0.5 mg/dL, and a high inflammatory status (HIS) group of 243 patients with a CRP of 0.5 mg/dL or more. Follow-up lasted a maximum of 3 years (median, 1.8 +/- 1.5 years) and was 92.6% complete. RESULTS: In-hospital mortality was 8.2% in the HIS group and 3.4% in the LIS group (odds ratio [OR], 2.61; p = 0.02). Incidence of postoperative infections was 16.5% in the HIS group and 5.1% in the LIS group (OR, 3.25; p = 0.0001). Sternal wound infections were also more frequent in the HIS group (10.7% versus 2.8%; OR, 3.43; p = 0.002). During follow-up, the HIS group had worse survival (88.5% +/- 2.9% versus 91.9% +/- 2.5%; OR, 1.93; p = 0.05) and a higher need of hospitalization for cardiac-related causes (73.6% +/- 6% versus 86.5% +/- 3.2%; OR, 1.82; p = 0.05). CONCLUSIONS: Patients undergoing cardiac surgery with a CRP level of 0.5 mg/dL or more are exposed to a higher risk of in-hospital mortality and postoperative infections. Despite surgical correction of cardiac disease, a high preoperative CRP value is an independent risk factor for mid-term survival and hospitalization for cardiac causes. PMID- 17126131 TI - Systemic stress hormone response in patients undergoing open heart surgery with or without cardiopulmonary bypass. AB - BACKGROUND: Cardiopulmonary bypass often causes a stress hormonal response with subsequent changes in hemodynamics and organ perfusion. Leptin, an adipocyte derived factor, has been proposed to play a role in systemic inflammation. We examined perioperative release of leptin and cortisol in patients undergoing open heart surgery with or without cardiopulmonary bypass. METHODS: Forty-nine patients were enrolled in this prospective study. Conventional coronary artery bypass grafting was performed in 19 patients (ONCABG; group I), and heart valve surgery in 15 patients (group II). Fifteen patients (group III) received off-pump coronary artery bypass grafting (OPCABG). Blood samples were collected preoperatively and for as long as 72 hours postoperatively. Plasma levels of leptin and cortisol were measured by enzyme-linked immunosorbent assay. RESULTS: Leptin serum levels decreased during the operation, reaching 73.2% of the baseline in group I, 85.3% in group II, and 38.9% in group III (p < 0.05), 2 hours postoperatively. Thereafter, leptin levels increased gradually to 218.6% of the baseline in group I and 313.7% in group II 24 hours after the operation (p < 0.01). However, patients in the OPCABG group showed only a moderate increase in serum leptin levels. Plasma cortisol levels rose to a maximum of 532.9% of baseline in group I, 526.4% in group II, and 280% in group III 12 hours postoperatively (p < 0.01). CONCLUSIONS: Open heart surgery is associated with acute perioperative changes in plasma levels of neurohormonal stress factors leptin and cortisol. A different pattern of leptin and cortisol release was observed in patients operated on without cardiopulmonary bypass. PMID- 17126132 TI - Invited commentary. PMID- 17126133 TI - Thrombophilia in cardiac surgery--patients with protein S deficiency. AB - BACKGROUND: Thrombophilic diathesis may cause severe problems in cardiac surgical patients. Among these, protein S deficiency is a coagulation disorder associated with recurrent thromboembolic events. We analyzed our experience with 7 patients with protein S deficiency who underwent cardiac surgery. METHODS: We retrospectively reviewed the clinical data, operative and postoperative courses, and the long-term results of 7 patients who were diagnosed to have protein S deficiency. Six of them were operated on using cardiopulmonary bypass, one was operated on with an off-pump procedure. RESULTS: Procedures performed were emergent pulmonary embolectomy (patient 1), aortic valve replacement and coronary artery bypass grafting (CABG, patient 2), re-CABG (patients 3 and 7), and CABG (patients 4, 5, and 6). In patients 1, 2, 3, and 7, the diagnosis was made perioperatively. Patients 4, 5, and 6 were treated with a modified regimen of warfarin or protamine. All of the latter 3 patients had an uneventful perioperative course without thromboembolic complication. At follow-up, all but 1 of the 7 patients were on continuous warfarin, and were well and without any further thromboembolic events. CONCLUSIONS: In patients with a past medical history of thromboembolic events or with a perioperative thromboembolic complication, elaborate laboratory investigation should lead to a definite diagnosis. For instance, patients with protein S deficiency undergoing cardiac surgery belong to a high-risk subgroup. Although rare, this and other coagulation disorders can be a critical issue in cardiac surgery. In such patients, we suggest perioperative warfarin therapy with a target international normalized ratio of 2.0 and incomplete protamine antagonism to minimize the risk of a perioperative thromboembolic event. PMID- 17126134 TI - Dopamine induces postischemic cardiomyocyte apoptosis in vivo: an effect ameliorated by propofol. AB - BACKGROUND: Dopamine is commonly used to improve postischemic myocardial contractile function. However, there is evidence that dopamine augments apoptosis after ischemia through increased intracellular calcium and opening of the mitochondrial permeability transition pore. Propofol (2,6-diisopropylphenol) is an anesthetic that has been shown to prevent mitochondrial permeability transition pore opening. We evaluated the effects of propofol given during reperfusion on dopamine-mediated apoptosis. METHODS: Hearts from 8-week-old inbred New Zealand White rabbit siblings were subjected to 2 hours of cold cardioplegic ischemia and 6 hours of reperfusion in a heterotopic transplant model. Controls consisted of the recipient rabbit's nonischemic heart. The ischemia-reperfusion (IR) group consisted of postischemic hearts reperfused with no drugs; the IR plus dopamine (IR+D) group received dopamine (20 microg x kg(-1) x min(-1)) continuously; the IR+D plus propofol (IR+D+P) group received dopamine (20 microg x kg(-1) x min(-1)) plus propofol (500 to 600 microg x kg(-1) x min( 1)); and the IR plus propofol (IR+P) group received propofol only (500 to 600 microg x kg(-1) x min(-1)) throughout reperfusion (n = 7 to 9 in each group). Myocardial function was measured using a left ventricular balloon; terminal nick end labeling (TUNEL) staining, DNA electrophoresis, and immunoblotting for caspase-3 cleavage were performed at the end of reperfusion. RESULTS: Dopamine increased the number of TUNEL-positive nuclei significantly (14.0 +/- 2.0/1,000 for IR+D versus 6.7 +/- 2.0/1,000 for IR, p = 0.01). Propofol (IR+D+P) reduced the total number of apoptotic cells in hearts receiving dopamine (7.1 +/- 1.8/1,000, p = 0.01 versus IR+D) to the extent seen in IR alone. DNA laddering and caspase-3 cleavage were observed at greater frequency in the IR+D group compared with the IR and IR+D+P groups. Propofol had no effect on dopamine mediated increased systolic function, but improved diastolic function after ischemia. CONCLUSIONS: Dopamine infusion has a positive inotropic effect on the postischemic heart at the expense of increased cardiomyocyte apoptosis. The addition of propofol prevents dopamine-induced apoptosis after ischemia while maintaining positive inotropy. PMID- 17126135 TI - Truncus arteriosus communis: early and midterm results of early primary repair. AB - BACKGROUND: Repair of truncus arteriosus communis (TAC) in the neonatal and early infant period has become standard practice in many centers. We report our experience on early primary repair of TAC, with a focus on early and midterm results. METHODS: From July 1993 to December 2005, 29 patients with median age 28 days (range, 11 to 127), and median body weight 3.1 kg (range, 2.6 to 5.9 kg), underwent primary repair of TAC. The anatomical type of TAC was as follows: A1-2, 27; A3, 0; and A4, 2. Right ventricular outflow tract was reconstructed with an aortic (n = 7) or pulmonary homograft (n = 8), or a bovine (n = 11) or porcine valved xenograft (n = 3). Follow-up was complete for all patients. RESULTS: Hospital mortality was 3.4% (1 death due to respiratory infection). At a mean follow-up of 74 months (range, 2 to 149), 1 patient died suddenly 2 months after surgery (6-year actuarial survival 93%). Of the 27 midterm survivors, 14 (52%) underwent 30 interventional procedures including percutaneous balloon dilation with or without stenting for right ventricular outflow tract or branch pulmonary artery obstruction. Eight of them were reoperated on for right ventricle-to pulmonary artery conduit replacement (n = 8, 23%), and aortic valve regurgitation (n = 1, 3.4%). The overall freedom from any reintervention at 6 years was 50%. Aortic valve regurgitation was trace in 15 patients, mild in 8, moderate in 4. All midterm survivors but 1 (26 of 27) had good ventricular function. CONCLUSIONS: Truncus arteriosus communis repair can be performed early with very low perioperative mortality and satisfactory midterm morbidity; the latter is mainly attributed to right ventricular outflow tract reconstruction. Interventional cardiac catheterization delays inevitable conduit replacement. PMID- 17126136 TI - Regional low-flow perfusion versus circulatory arrest in neonates: one-year neurodevelopmental outcome. AB - BACKGROUND: Regional low-flow perfusion of the brain is a bypass technique commonly used during stage 1 reconstruction in neonates with hypoplastic left heart syndrome and related variants. The neurodevelopmental outcome of these children is unknown. METHODS: Twenty-nine infants (22 boys, 7 girls) with hypoplastic left heart syndrome or variant requiring single ventricle palliation and aortic arch reconstruction were studied between 1999 and 2004. Mental Developmental Index (MDI) and Psychomotor Developmental Index were assessed using Bayley Scales of Infant Development and correlated with intraoperative and perioperative variables. Results are reported as mean +/- standard deviation. RESULTS: Average age at stage 1 operation and at bidirectional Glenn was 7 +/- 8 days and 6.0 +/- 2 months, respectively. The MDI was in the low average range (87.7 +/- 13.2). The Psychomotor Developmental Index was in the mildly delayed range (75.2 +/- 14.5). Regional low-flow perfusion was used in 31% (9 of 29 patients), with an average circulatory arrest time of 23.5 +/- 13.4 minutes. Deep hypothermia and circulatory arrest was used as the primary operative strategy in 69% of patients (20 of 29 patients), with an average circulatory arrest time of 44.3 +/- 15.3 minutes (p = 0.003). No differences in MDI or Psychomotor Developmental Index scores were observed between the regional low-flow perfusion and non-regional low-flow perfusion groups (MDI, 88.0 +/- 12.1 versus 87.6 +/- 14.0; p = 0.93, respectively; Psychomotor Developmental Index, 75.5 +/- 15.1 versus 75.0 +/- 14.6; p = 0.93, respectively). Lowest operative temperature (<16 degrees C) and birth order (<2 versus >3) significantly related to MDI (89.6 versus 72.8; p = 0.047). CONCLUSIONS: At 1 year of age, neurodevelopmental outcomes of patients undergoing stage 1 using regional low-flow perfusion were similar to outcomes observed in children exposed to circulatory arrest. The association of birth order and MDI suggests that early intervention may benefit these patients. PMID- 17126137 TI - Pulmonary atresia with intact ventricular septum: initial management. AB - BACKGROUND: Outcomes for pulmonary atresia with intact ventricular septum are suboptimal, while initial management remains controversial. This study was undertaken to determine the effect of catheter-based therapy on the need for early surgical intervention. METHODS: A single-institution retrospective chart review was made of all 25 neonates with pulmonary atresia with intact ventricular septum from 1999 to 2005. RESULTS: Mean age at first intervention was 3.1 +/- 2.2 days, mean weight 3.3 +/- 0.5 kg. Right ventricular hypoplasia varied: 20% normal, 16% mild, 28% moderate, 28% moderately severe or severe, 8% not classified. Median tricuspid valve z-score was -2.3 +/- 2.6. First intervention was catheter-based therapy in 56% (14 of 25), operative in 36% (9 of 25), and no therapy in 2. Technically adequate valvotomy was achieved in 79% (11 of 14). Serious catheter-related complications occurred in 3 of 14 (21%). Only 5 of 14 (36%) with catheter-based therapy weaned from prostaglandins without surgery. Of these, 2 required surgery for cyanosis at 1 and 3 months. Surgery after catheter based therapy consisted of right ventricular outflow patch in 36% (4 of 11), systemic to pulmonary shunt in 64% (7 of 11). Median time between catheter-based therapy and surgery was 8.5 days (range, 1 to 89). Only 3 of the 23 treated patients avoided operation during infancy. There was 1 early and 1 late death after operation after initial catheter-based therapy, and 1 late death after primary surgery alone during a mean follow-up of 33 months (range, 1.5 to 79). CONCLUSIONS: Balloon valvotomy alone for pulmonary atresia with intact ventricular septum rarely obviates the need for an additional source of pulmonary blood flow--either shunt or ductal stenting. PMID- 17126138 TI - Surgical treatment of absent pulmonary valve syndrome associated with bronchial obstruction. AB - BACKGROUND: Absent pulmonary valve syndrome (APVS) is a relatively rare anomaly that is usually associated with a ventricular septal defect and a restrictive pulmonary annulus with severe pulmonary regurgitation. The to-and-fro flow across the right ventricular outflow tract produces enormously dilated and pulsatile pulmonary arteries (PAs) that cause severe respiratory distress and tracheomalacia by compression of the trachea and primary bronchi. This retrospective study summarizes our 20-year experience of surgical treatment in patients with APVS. METHODS: Between 1984 and 2005, 20 patients underwent repair of APVS using a valved conduit (n = 9), transannular patch (TAP) insertion alone (n = 5), or TAP with monocusp valve (n = 6) with PA reduction arterioplasty. Median age was 7 months (range, 6 days to 9 years). RESULTS: There were one perioperative and two late deaths. All deaths were related to airway complications. Survival was 90% at 1 year and 85% at 10 and 15 years. In a multivariate analysis only preoperative ventilator dependency was associated with a worse outcome (p = 0.02). Follow-up was available in 18 operative survivors (mean follow-up, 7.3 +/- 5.2 years). Six patients (33%) underwent reoperation for pulmonary valve incompetence and right ventricular dysfunction. Three patients (17%) had episodic bronchospasm of mild to moderate severity that were responsive to sympathomimetic bronchodilators. CONCLUSIONS: Morbidity associated with perioperative respiratory complications and ventilator dependency due to underlying tracheobronchomalacia is an important problem in patients with APVS. These infants may require multiple hospitalizations for recurrent respiratory infections secondary to their tracheobronchomalacia. Complete repair with a valved conduit and reduction pulmonary arterioplasty at the onset of symptoms and a definitive diagnosis is our procedure of choice for infants with APVS. With this approach, the airway can be optimized to give the best patient outcome. PMID- 17126139 TI - Aortic atresia or severe left ventricular outflow tract obstruction with ventricular septal defect: results of primary biventricular repair in neonates. AB - BACKGROUND: Aortic atresia or severe aortic stenosis and left ventricular outflow tract obstruction is a frequent component of complex congenital heart disease. Aortic atresia or severe aortic stenosis and left ventricular outflow tract obstruction with two adequate ventricles is sometimes treated by Norwood palliation followed by late biventricular repair. We reviewed our experience with primary biventricular repair in this group of neonates. METHODS: Retrospective review identified 17 neonates (10 males) with aortic atresia or severe left ventricular outflow tract obstruction with ventricular septal defect and an adequate left ventricle undergoing primary biventricular repair between 1986 and 2002. Mean age was 7.7 +/- 2.9 days, weight 3.3 +/- 0.7 kg, and body surface area 0.21 +/- 0.04 kg/m2. Associated anomalies included arch hypoplasia, 7 (41%); aortic atresia, 7 (41%); and coarctation, 5 (29%). Results are reported as mean +/- standard deviation. RESULTS: Median follow-up was 6 years (range, 1 to 17.7 years). Three of the 17 (18%) died within 30 days. There were no deaths in this series since 1992. Nine patients (38.9%) required one reoperation, 7 of which were for conduit stenosis, 1 for left ventricular outflow tract obstruction, and 1 for residual ventricular septal defect with left ventricle-to-right atrium shunt. Freedom from death at 10 years was 82% by Kaplan-Meier estimate. CONCLUSIONS: Excellent long-term survival can be achieved by primary biventricular repair as corroborated by our survival rate of 82%. Primary biventricular repair is an effective operation for aortic atresia and severe left ventricular outflow tract obstruction with adequate sized left ventricle that avoids interstage attrition associated with Norwood palliation and is our procedure of choice. PMID- 17126140 TI - Moderately hypothermic cardiopulmonary bypass and low-flow antegrade selective cerebral perfusion for neonatal aortic arch surgery. AB - BACKGROUND: Although deep hypothermic circulatory arrest has been extensively used in neonates for aortic arch surgery, the brain and other organs might be adversely affected by prolonged ischemia and deep hypothermia. METHODS: Between December 1997 and January 2005, 70 consecutive neonates underwent Norwood stage I procedure for hypoplastic left heart syndrome (group A, n = 30), or aortic arch repair for interruption or coarctation with arch hypoplasia (group B, n = 40), with antegrade selective cerebral perfusion (ASCP). Mean weights were 3.0 +/- 0.2 kg and 2.8 +/- 0.07 kg, and mean ages were 10 +/- 3.5 days and 14 +/- 10.6 days in groups A and B, respectively. Only 2 patients were older than 30 days. Core body temperature was lowered to 25 degrees C, and mean pump flow during ASCP was initiated at 10 to 20 mL/(kg x min) and adjusted to guarantee a radial/temporal artery pressure of 30 to 40 mm Hg and venous oxygen saturation of more than 70%. Hematocrit was maintained at 30%. RESULTS: Early mortality was 17% (group A, 23%; group B, 12.5%; p = 0.19). Six late deaths occurred (3 in each group), and at 36 months, Kaplan-Meier overall survival was 64% +/- 9.2% in group A and 85% +/- 5.7% in group B. One patient had postoperative seizures. Age, weight, sex, prematurity, group A, and ASCP duration did not influence early mortality. CONCLUSIONS: Antegrade selective cerebral perfusion is a safe and effective procedure and might improve outcome of neonatal aortic arch surgery, minimizing neurologic impact without the need for deep hypothermia. PMID- 17126141 TI - Discrepancy between intraoperative transesophageal echocardiography and postoperative transthoracic echocardiography in assessing congenital valve surgery. AB - BACKGROUND: The purpose of this study was to investigate the discrepancy between intraoperative transesophageal and postoperative transthoracic echocardiography in assessing residual regurgitation in children undergoing valve repair. METHODS: Forty-two consecutive children (median age, 5.1 years) who underwent valve repair for valvar regurgitation from 2001 to 2004 were retrospectively analyzed. The patients were divided into two groups: atrioventricular valve group (n = 33) and aortic valve group (n = 9). Regurgitation grade, fractional shortening, and atrioventricular inflow velocity obtained by intraoperative transesophageal echocardiography were compared with those obtained by transthoracic echocardiography at discharge (median, 11 days) and at follow-up (median, 8 months). RESULTS: Intraoperative transesophageal echocardiography revealed specific residual lesions in 4 patients, leading to successful re-repair. Fractional shortening obtained by intraoperative transesophageal echocardiography was lower than that obtained by predischarge transthoracic echocardiography (p < 0.01). In the atrioventricular valve group, the regurgitation grade obtained by intraoperative transesophageal echocardiography was lower than that obtained by predischarge transthoracic echocardiography (0.7 +/- 0.8 versus 1.4 +/- 0.9; p < 0.01), and agreement between the two examinations was found in 12 patients (38%). Peak atrioventricular inflow velocity obtained by intraoperative transesophageal echocardiography was lower than that obtained by predischarge transthoracic echocardiography (p < 0.01). In the aortic valve group, there was no significant difference between the regurgitation grades in the two examinations (0.8 +/- 0.8 versus 1.1 +/- 0.9), and complete agreement in regurgitation grade was found in 5 (56%) of 9 patients. CONCLUSIONS: There were considerable discrepancies between the examinations in evaluation of residual atrioventricular valve regurgitation and potential atrioventricular valve stenosis: most of the residual regurgitations were underestimated by intraoperative transesophageal echocardiography. In contrast, reasonable agreement was found between the two examinations in evaluation of aortic valve regurgitation. PMID- 17126142 TI - Regulation of brain cell death and survival after cardiopulmonary bypass. AB - BACKGROUND: This study investigated the effect of low flow cardiopulmonary bypass, circulatory arrest, and selective cerebral perfusion on expression and phosphorylation of selected regulators of cell death and survival in striatum of newborn piglets. METHODS: Animals were assigned to sham operation and three experimental groups. The experimental groups were placed on bypass, cooled to 18 degrees C, and subjected to 90 minutes of deep hypothermic circulatory arrest (DHCA), low-flow cardiopulmonary bypass (LFCPB) at mL/(kg x min), or selective cerebral perfusion (SCP) at 20 mL/(kg x min), followed by rewarming and 2 hours of recovery. The oxygen pressure in the microcirculation of the cortex was measured by quenching of phosphorescence. Levels of phosphorylated and total protein were determined by Western blot analysis. RESULTS: Control oxygen pressure was 55 +/- 9 mm Hg and decreased during DHCA, LFCPB, and SCP to 1.1 +/- 0.6 mm Hg, 9.8 +/- 2.3 mm Hg, and 9.3 +/- 1.9 mm Hg, respectively (p < 0.001). After DHCA, N-terminal of Bcl-2-associated X protein (N-Bax) levels increased (295% +/- 15%, p < 0.01), B-cell leukemia protein (Bcl-2) levels decreased (31% +/- 9%, p < 0.01), and phosphorylation level of protein kinase B (pAkt) and extracellular signal-regulated kinase 1/2 (pERK1/2) did not change. After LFCPB and SCP, N-Bax and Bcl-2 levels were unchanged, pAkt levels increased (367% +/- 122%, p < 0.05 and 337% +/- 47%, p < 0.01, respectively), pERK1 (484% +/- 70% and 501% +/- 255%, respectively; p < 0.01) and pERK2 (569% +/- 128%; p < 0.001 and 494% +/- 162%; p < 0.05, respectively) levels increased, and total ERK2 levels also increased (279% +/- 90% and 153% +/- 44%, respectively, p < 0.05). CONCLUSIONS: Stable levels of Bcl-2 and Bax and the increases in pAkt and pERK1/2 after LFCPB and SCP are likely indicators of improved chances for cell survival. PMID- 17126143 TI - Epicardial real-time three-dimensional echocardiography in cardiac surgery: a preliminary experience. AB - PURPOSE: Intraoperative two-dimensional transesophageal echocardiography (2DTEE) is a widely accepted method to guide cardiac valve surgery. The aim of our study was to evaluate the feasibility, effectiveness, and incremental value of intraoperative epicardial real-time three-dimensional echocardiography (RT3DE). DESCRIPTION: Thirty consecutive patients (18 aortic and 12 mitral valve diseases) underwent intraoperative 2DTEE and RT3DE before and after cardiopulmonary bypass. Five observers compared independently 2DTEE to live and full volume images and to the surgical view, to assess the incremental value of RT3DE in depicting the different anatomic structures. EVALUATION: Epicardial RT3DE was feasible in all patients. Qualitative evaluation determined RT3DE superiority in depicting aortic cusp morphologic lesions; left ventricular outflow tract spatial relationships with mitral apparatus and aortic root; and both anterior and posterior mitral leaflet scallops, particularly posterior commissure. CONCLUSIONS: In our study, epicardial RT3DE has been demonstrated to improve morphologic definition of anatomic valvular lesions and their relationship with cardiac adjacent structures. It may be a valid substitute when the 2DTEE approach is contraindicated, or it could have a complementary role, coupled with 2DTEE, to give additional information for surgical planning. PMID- 17126144 TI - Laser energy source in surgical atrial fibrillation ablation: preclinical experience. AB - PURPOSE: The purpose of this study was to evaluate diffusing tip laser energy in surgical atrial fibrillation ablation using a canine model. This is the first report to describe the pathological and histological findings using a laser energy source. DESCRIPTION: The surgical atrial fibrillation ablation procedure was performed through a left atriotomy; the pulmonary veins were encircled in 16 dogs using a diode laser (980 nm) with a diffusing tip that permits linear ablation perpendicular to the fiber direction. Lesion durations were 45 seconds with a power density of 3.8 or 4.5 W/cm. Six animals were allowed to survive 4 hours after the procedure, with the remainder sacrificed at 1 week (n = 1), 4 weeks (n = 3), and 6 weeks (n = 6). Electrophysiologic effectiveness was judged using unipolar or bipolar pacing from the pulmonary veins after attempting isolation. Hearts were harvested for histologic examination using standard trichrome staining. EVALUATION: All animals tolerated the procedure. The animals required an average of 5.6 +/- 0.82 lesions to complete the procedure. All animals had confirmed isolation of the pulmonary veins as judged by unipolar or bipolar pacing, and this isolation persisted in those animals that were allowed to survive. Pathology revealed all lesions to be transmural with an average tissue thickness of 3.62 +/- 1.50 mm (range, 0.95 mm to 7.06 mm). CONCLUSIONS: Diffusing tip laser technology reproducibly makes rapid, transmural, and electrophysiologically effective atrial lesions. PMID- 17126145 TI - Invited commentary. PMID- 17126146 TI - Typical bronchopulmonary carcinoid tumors: a ramifying bronchial presentation with metastatic behavior. AB - Bronchopulmonary typical carcinoid tumors (BTCT) are neuroendocrine neoplasms with histologic low grade characteristics considered benign. However, despite reassuring histologic classification, some of them demonstrate an aggressive nature and metastatic behavior. During a not yet concluded study aiming at establishing criteria to predict this metastatic behavior, three uncommon cases were observed. Metastasis occurred despite typical carcinoid microscopic features in 3 female patients of African origin presenting at macroscopic examination as ramifying bronchopulmonary typical carcinoid tumors following the bronchial tree. We suggest that clinical ramifying presentation may be related to metastatic behavior, even for bronchopulmonary typical carcinoid tumors not displaying histologic criteria for atypical carcinoid tumors. PMID- 17126147 TI - Thymic small cell carcinoma associated with pulmonary squamous cell carcinoma. AB - A 63-year-old man presented with dyspnea on effort. Chest computed tomography showed an anterior mediastinal mass and a lung mass in the right lower lobe. Thallium scintigraphy revealed accumulation in the mediastinal mass. Therefore, under diagnosis of invasive thymoma or thymic carcinoma associated with suspected lung cancer, exploratory right thoracotomy was undertaken through a median sternotomy with video-assisted thoracoscopic support. The lung mass was intraoperatively diagnosed as squamous cell carcinoma. Right lower lobectomy and total thymectomy were then carried out without additional incision. Thymic small cell carcinoma was diagnosed; therefore the patient received 50 Gy of irradiation to the mediastinum. Ten months after surgery the patient is alive without recurrence. PMID- 17126148 TI - Treatment of bronchomalacia in cystic fibrosis by silicone stent. AB - We report the case of a 15-year-old girl with cystic fibrosis and rapidly declining pulmonary function tests who was found to have collapse of the left main bronchus from bronchomalacia. She underwent successful deployment of an expandable silicone stent in the collapsed bronchus, after which her pulmonary function test results and her clinical picture markedly improved, obviating the need for immediate transplantation. A literature review yielded no prior reports of bronchomalacia in a cystic fibrosis patient being treated with a silicone stent. This case shows that a simple, effective treatment is possible for one cause of obstructive pulmonary function in cystic fibrosis. PMID- 17126149 TI - Pulmonary artery aneurysm with ascending aortic aneurysm concomitant with bilateral bicuspid semilunar valves. AB - Aneurysm of both the pulmonary trunk and the ascending aorta concomitant with bilateral bicuspid valves is very rare. The reason for the formation of aneurysm with bicuspid semilunar valve is still inconclusive. Surgical repair was performed successfully (ie, aortic valve replacement, graft replacement for the ascending aorta, and plication of the pulmonary artery). Pathology of the pulmonary artery wall did not demonstrate cystic medial necrosis. The hemodynamic turbulence by the bilateral bicuspid valve may cause the formation of aneurysms even at low pressure. This case demonstrates an explanation for aneurysm with the bicuspid valve. PMID- 17126150 TI - Anomalous right upper lobe venous drainage. AB - Anomalous pulmonary venous return is a rare congenital anomaly mainly involving the right lung and is often associated with congenital intracardiac malformations as atrial septal defect. We report a case of anomalous right upper lobe venous drainage resulting in two right upper lobe veins draining into the azygous vein and into the confluence between superior vena cava and azygous vein, respectively. Preoperative identification of such an aberrant venous drainage is useful for avoiding unexpected intraoperative bleeding. PMID- 17126151 TI - Intimo-intimal intussusception of the aorta. AB - A 50-year-old man presented with acute onset of chest pain. Subsequent transesophageal echocardiography and computed tomography scan showed absence of a flap in the ascending aorta and a clear dissection flap involving the arch and descending aorta. Magnetic resonance imaging showed a tear and a small flap in the right coronary sinus. During surgery, we found a total circumferential intimal tear at the sinotubular junction with intimo-intimal intussusception of the internal channel into the arch. Dissection without intimal flap and aortic intussusception is a rare form of type A dissection, which is difficult to diagnose on routine investigations and can delay treatment. PMID- 17126152 TI - Endovascular treatment of an acute subdiaphragmatic aortic rupture. AB - We report the case of a 23-year-old man presenting an acute rupture of the subdiaphragmatic aorta in front of a T12 vertebral fracture after a road accident. Because of the location of this lesion, the operative risk and a cardiac instability, we opted for an endovascular treatment with a new and original approach in covering a small part of the aorta using commercial devices. We also describe the probable mechanism of this uncommon aortic rupture. The surgical outcome was uneventful and the 3 month computed tomographic scan confirmed the complete exclusion of the aortic disruption. PMID- 17126153 TI - Erosion of an esophageal endoprosthesis into the trachea. AB - Acquired benign non-neoplastic tracheoesophageal fistulas are unusual, demanding clinical entities. Surgical technique and final outcome depends on fistula size. A one-stage procedure with esophageal resection is preferred when the fistula is located in the upper third of the trachea and is less than 1 cm long. PMID- 17126154 TI - Pseudoaneurysm of an aortic homograft. AB - False aneurysms of the thoracic aorta constitute one of the most challenging problems encountered by the cardiac surgeon. We report a case of successful reoperation for a giant pseudoaneurysm of an aortic homograft, previously used in the context of postpartum acute endocarditis of the aortic valve. PMID- 17126155 TI - Perforation of the Valsalva sinus after implantation of Medtronic Freestyle aortic bioprosthesis. AB - We report on structural valve deterioration in patients with the Medtronic Freestyle aortic bioprosthesis (Medtronic, Inc, Minneapolis, MN), including spontaneous perforation of the Valsalva sinus. These occurred in four prosthesis in 3 patients using the modified subcoronary method or full root technique. One patient died of ruptured pseudoaneurysm and the others survived reoperation well. Careful follow-up is required after Freestyle bioprosthesis implantation. PMID- 17126156 TI - Anomalous left main coronary artery from the main pulmonary artery in an elderly patient. AB - Anomalous origin of the left main coronary from the pulmonary trunk in an elderly patient is extremely rare. We report a 73-year-old woman who presented with new onset of angina and atrial fibrillation. Evaluation revealed anomalous origin of the left main coronary artery from the main pulmonary artery and tight proximal left anterior descending coronary artery stenosis. The patient underwent primary closure of the anomalous left main coronary artery orifice within the pulmonary artery, aorta-to-left anterior descending coronary artery saphenous vein bypass grafting and Maze procedure. Six months postoperatively the patient was asymptomatic and a Thallium stress test was negative for ischemia. PMID- 17126157 TI - Mitral valve cleft in both anterior and posterior leaflet: an extremely rare anomaly. AB - We herein report the case of a woman with clefts in the anterior and posterior mitral leaflets causing mitral insufficiency. The patient developed heart failure symptoms in the eighth decade of life and underwent mitral valve repair, with suture closure of both clefts. The reasons of its late presentation are discussed. PMID- 17126158 TI - Transesophageal echocardiography guidance for repair of complex cardiac injuries. AB - Our patient suffered a single stab wound to the heart that resulted in a coronary artery to ventricle fistula, a ventricular septal defect, mitral regurgitation from chordal transection, and a delayed left ventricular aneurysm. We used transesophageal echocardiography to characterize these multiple defects to help guide successful surgical repair. The patient recovered uneventfully. PMID- 17126159 TI - Left ventricular pacing site-timing optimization during biventricular pacing using a multi-electrode patch. AB - A 71-year-old man with class IV congestive heart failure and an infected pacemaker/implantable cardioverter defibrillator (ICD) underwent median sternotomy for removal of endocardial leads with a 15-mm vegetation. Cardiac output during biventricular pacing was optimized with an aortic flow probe, a multi-electrode left ventricular patch, and a randomized protocol assessing 54 combinations of pacing site and right ventricle-left ventricle delay. Results that were assessed with response surface methodology determined permanent epicardial lead position and timing. The difference between the best and worst site-timing combinations altered cardiac index by nearly 70%. This experience demonstrates potential importance of the epicardial approach to site-timing optimization for biventricular pacing. PMID- 17126160 TI - Left ventricular posterior wall pseudoaneurysm: a rare sequela of mitral valve infective endocarditis in a chronic patient of HLA-B27 positive spondyloarthritis. AB - Left ventricular posterior wall pseudoaneurysm after native mitral valve infective endocarditis is a very rare occurrence. We report such a case in a patient with HLA-B27-associated spondyloarthritis and normal coronary arteries. Excision of the aneurysm with left ventricular reconstruction and mitral valve replacement resulted in an excellent outcome. PMID- 17126161 TI - Cystic formation of the foramen ovale mimicking a right atrial myxoma. AB - A 65-year-old woman presented with shortness of breath, stenocardia, and tachycardia. She underwent several steps of examination. Echocardiography showed a suspicious formation in the right atrium extending into the inferior caval vein. The lesion was suspicious for a myxoma, a thrombus, or a malformation. Intraoperatively this formation presented as a cystic formation connected to the right atrial wall in the area of the foramen secundum and filled with blood and five calcified thrombi. We hypothesize that a slit-like opening in the foramen ovale produced a valve-like mechanism bulging parts of the septum secundum and produced this cystic formation. PMID- 17126162 TI - Left common carotid artery as inflow site in coronary artery bypass grafting. AB - Porcelain aorta is associated to significantly increased risk of atheromatous embolization in patients who undergo cardiac surgery. We described three cases in which coronary artery bypass grafting was performed off-pump and the saphenous vein graft was anastomosed proximally to the left common carotid artery. This technique permitted avoidance of ascending aortic manipulation and reduce the risk of atheromatous embolization. PMID- 17126163 TI - Stage I palliation for interrupted "right" aortic arch associated with mitral atresia. AB - A 2.6 kg female with the diagnosis of type "B" interruption of the "right" aortic arch and mitral atresia underwent stage I palliation with translocation and anastomosis of the distal right carotid artery to the descending aorta. The distal main pulmonary artery was anastomosed to the under surface of the neoaortic arch. A modified Blalock-Taussig shunt was constructed between the left side innominate artery and the left pulmonary artery. The child awaits stage II palliation. PMID- 17126164 TI - Giant metastatic endometrial sarcoma to the right lung. PMID- 17126165 TI - An extrapericardial middle mediastinal pheochromocytoma. PMID- 17126166 TI - Alternative approach to the surgical management of congenital tracheal stenosis. AB - Congenital tracheal stenosis is a serious disease associated with high operative mortality and morbidity, especially in the newborn period. Its surgical management involves either sliding tracheoplasty, which is associated with extensive dissection and recurrent stenosis, or with pericardial patch tracheoplasty that fails to address recoiling of the stenotic complete ring and collapse of the patch into the tracheal lumen. We report a modification of the latter technique, in which a half ring external stenting device is used to keep the rings from recoiling. PMID- 17126167 TI - Decompression of giant bulla in acute pneumonia: surgical palliation prior to definitive management. AB - Giant bullae can be complicated by respiratory tract infection in the setting of emphysema. Herein we describe a technique of palliative decompression of the bullae that gives time to treat acute pulmonary infection prior to definitive surgical treatment. PMID- 17126168 TI - Bridging the cleft over the throbbing heart. AB - Primary repair of sternal cleft deformities are best achieved in the neonatal period. Conversion of a partial defect to a complete sternal cleft with sternal bar mobilization is able to achieve sternal approximation in most cases. We describe a new technique of fracturing the clavicles in unyielding cases as was our experience in a 6-week-old infant. PMID- 17126169 TI - Teflon felt wrapping repair for coronary perforation after failed angioplasty. AB - Acute type III perforation caused by failed angioplasty is a lethal complication that often requires emergency operation. However, the presence of multiple rigid stents beneath the subepicardial hematoma disturbs optimal revascularization and hemostasis. Teflon felt (Meadox Medical Inc, Oakland, NJ) wrapping repair is a simple salvage technique that allows stable hemostasis and the rescue of the entire blood flow of the coronary artery. This procedure was successfully performed with type III perforation of the left anterior descending coronary artery on 2 patients subjected to multiple stenting. PMID- 17126170 TI - Thrombin during cardiopulmonary bypass. AB - Cardiopulmonary bypass (CPB) ignites a massive defense reaction that stimulates all blood cells and five plasma protein systems to produce a myriad of vasoactive and cytotoxic substances, cell-signaling molecules, and upregulated cellular receptors. Thrombin is the key enzyme in the thrombotic portion of the defense reaction and is only partially suppressed by heparin. During CPB, thrombin is produced by both extrinsic and intrinsic coagulation pathways and activated platelets. The routine use of a cell saver and the eventual introduction of direct thrombin inhibitors now offer the possibility of completely suppressing thrombin production and fibrinolysis during cardiac surgery with CPB. PMID- 17126171 TI - Indications for blood transfusion in cardiac surgery. AB - In addition to its life-saving effect in hemorrhagic shock, transfusion of allogenic packed red blood cells can be beneficial in situations where a critically low hematocrit is contributing to a state of oxygen-supply dependency. These benefits are countered by the risks of transfusion-associated lung injury, transfusion-associated immunomodulation, and cellular hypoxia after RBC transfusion. The critical hematocrit is patient and organ specific, and varies intraoperatively according to the duration and temperature of bypass, as well as for a variable postoperative period. Future randomized studies must prospectively evaluate regional indicators of tissue oxygenation in transfusion algorithms. PMID- 17126172 TI - Staging in malignant mesothelioma. PMID- 17126174 TI - Should pleural fluid cytology be incorporated in the TNM staging system for operable nonsmall cell lung cancer? PMID- 17126176 TI - Pulmonary valve implantation with a self-expanding stented valve: 1-year follow up in the first patient. PMID- 17126177 TI - Durability of valve bioprostheses and the need of careful examination of explants. PMID- 17126178 TI - Coronary artery reconstruction: optimal technique of coronary endarterectomy. PMID- 17126180 TI - Compensation and egg donation for research. AB - Financial compensation to egg donors for their time and inconvenience in donating is established in the United States as ethically and legally acceptable. That system should be extended to egg donation for research. PMID- 17126182 TI - Peace. PMID- 17126183 TI - Sometimes they live. PMID- 17126184 TI - Response to Molczan. Cardiac anomalies in the neonate: high index of suspicion important. PMID- 17126185 TI - Emergency nurses' perceptions of obstacles and supportive behaviors in end-of life care. AB - INTRODUCTION: Little is known about emergency nurses' perceptions of either obstacles or supportive behaviors for providing end-of life (EOL) care to dying patients. The purpose of this study was to determine the perceived obstacles and supportive behaviors in providing EOL care to dying patients in emergency departments. METHODS: In this survey research, a 73-item questionnaire regarding EOL care was mailed to a geographically dispersed national random sample of 300 members of the Emergency Nurses Association. Descriptive statistics were calculated for the 54 Likert-type items and demographic items. Two open-ended questions were analyzed using content analysis. RESULTS: Returns after 3 mailings yielded 169 usable questionnaires from 284 eligible respondents for a return rate of 59.5%. The greatest obstacles were: (1) emergency nurses having too great a work load to care for dying patients; (2) emergency nurses having to deal with angry family members; and (3) the poor design of emergency departments that do not allow for privacy of dying patients or grieving family members. The most supportive behaviors were: (1) good communication between the physician and RN caring for the dying patient; (2) physicians meeting in person with the family after the patient's death; and (3) an emergency department designed so that the family has a place to grieve in private. DISCUSSION: Having a better understanding of emergency nurses' perceptions of obstacles and supportive behaviors in providing end-of-life care could help decrease the stress of caring for dying patients. Actions could be taken to decrease the highest rated obstacles and increase the ratings of supportive behaviors that may ultimately result in better end-of-life care for dying patients and their families in the emergency setting. PMID- 17126186 TI - Interhospital transfer for primary percutaneous coronary intervention: a community hospital's experience. PMID- 17126187 TI - Development and implementation of a patient safety program in an academic, urban emergency department. PMID- 17126188 TI - Utilization of information systems for ED disaster registration and tracking. PMID- 17126189 TI - Creating an enhanced triage area improves emergency department throughput. PMID- 17126190 TI - A 23-year-old woman with cystic fibrosis, acute hypotension, and cardiac arrest. PMID- 17126192 TI - Online nursing skills. PMID- 17126193 TI - Caring for the patient with a spontaneous abortion. PMID- 17126194 TI - Medication reconciliation: doing what's right for safe patient care. PMID- 17126195 TI - Ballistic injury to the chest. PMID- 17126196 TI - Preventing staff injuries from violence. PMID- 17126198 TI - Annual emergency department competencies: a non-traditional approach. PMID- 17126199 TI - The crying infant in the emergency department. PMID- 17126200 TI - The risky business of body packers and body stuffers. PMID- 17126201 TI - Universal health care at the local level: can reform increase access and help our emergency care system? PMID- 17126202 TI - Blunt laryngeal trauma. PMID- 17126203 TI - Radiation pneumonitis and fibrosis: mechanisms underlying its pathogenesis and implications for future research. AB - Radiation pneumonitis and subsequent radiation pulmonary fibrosis are the two main dose-limiting factors when irradiating the thorax that can have severe implications for patients' quality of life. In this article, the current concepts about the pathogenetic mechanisms underlying radiation pneumonitis and fibrosis are presented. The clinical course of fibrosis, a postulated acute inflammatory stage, and a late fibrotic and irreversible stage are discussed. The interplay of cells and the wide variety of molecules orchestrating the immunologic response to radiation, their interactions with specific receptors, and the cascade of events they trigger are elucidated. Finally, the implications of this knowledge with respect to the therapeutic interventions are critically presented. PMID- 17126204 TI - Radiation proctopathy in the treatment of prostate cancer. AB - PURPOSE: To compile and review data on radiation proctopathy in the treatment of prostate cancer with respect to epidemiology, clinical manifestations, pathogenesis, risk factors, and treatment. METHODS: Medical literature databases including PubMed and Medline were screened for pertinent reports, and critically analyzed for relevance in the scope of our purpose. RESULTS: Rectal toxicity as a complication of radiotherapy has received attention over the past decade, especially with the advent of dose-escalation in prostate cancer treatment. A number of clinical criteria help to define acute and chronic radiation proctopathy, but lack of a unified grading scale makes comparing studies difficult. A variety of risk factors, related to either radiation delivery or patient, are the subject of intense study. Also, a variety of treatment options, including medical therapy, endoscopic treatments, and surgery have shown varied results, but a lack of large randomized trials evaluating their efficacy prevents forming concrete recommendations. CONCLUSION: Radiation proctopathy should be an important consideration for the clinician in the treatment of prostate cancer especially with dose escalation. With further study of possible risk factors, the advent of a standardized grading scale, and more randomized trials to evaluate treatments, patients and physicians will be better armed to make appropriate management decisions. PMID- 17126205 TI - Delayed breast cellulitis: an evolving complication of breast conservation. AB - PURPOSE: Delayed breast cellulitis (DBC) is characterized by the late onset of breast erythema, edema, tenderness, and warmth. This retrospective study analyzes the risk factors and clinical course of DBC. METHODS AND MATERIALS: From 1985 through 2004, 580 sequential women with 601 stage T0-2N0-1 breast cancers underwent breast conserving therapy. Cases of DBC were identified according to accepted clinical criteria: diffuse breast erythema, edema, tenderness, and warmth occurring >3 months after definitive surgery and >3 weeks after radiotherapy. Potential risk factors analyzed included patient comorbidity, operative technique, acute complications, and details of adjunctive therapy. Response to treatment and long-term outcome were analyzed to characterize the natural course of this syndrome. RESULTS: Of the 601 cases, 16%, 52%, and 32% were Stage 0, I, and II, respectively. The overall incidence of DBC was 8% (50/601). Obesity, ecchymoses, T stage, the presence and aspiration of a breast hematoma/seroma, removal of >5 axillary lymph nodes, and arm lymphedema were significantly associated with DBC. The median time to onset of DBC from the date of definitive surgery was 226 days. Ninety-two percent of DBC patients were empirically treated with antibiotics. Fourteen percent required more invasive intervention. Twenty-two percent had recurrent episodes of DBC. Ultimately, 2 patients (4%) underwent mastectomy for intractable breast pain related to DBC. CONCLUSION: Although multifactorial, we believe DBC is primarily related to a bacterial infection in the setting of impaired lymphatic drainage and may appear months after completion of radiotherapy. Invasive testing before a trial of antibiotics is generally not recommended. PMID- 17126206 TI - Multi-institutional study of radiation therapy for isolated para-aortic lymph node recurrence in uterine cervical carcinoma: 84 subjects of a population of more than 5,000. AB - PURPOSE: Most patients who had any recurrent sites of cancer have been considered to be in their last stage of life. However, recent advances of clinical research reveal some patients achieve long-term survival even in recurrence. Furthermore, for patients who had only one recurrent region, radiation therapy could play an important role. As for uterine cervical carcinoma, the most common recurrent site other than the pelvis is the para-aortic lymph nodes. Thus we conducted the current study. PATIENTS AND METHODS: Between 1994 and 2003, more than 5,000 uterine cervical carcinoma patients were treated with curative intended treatments at 13 Japanese hospitals. Of these patients, 84 developed para-aortic lymph node recurrence as the only site of initial tumor progression. These patients were treated with external beam radiation therapy. Radiation therapy protocol was as follows: 1.7-2.0 Gy per fraction, 5 fractions per week, and the mean total dose was 50.8 Gy (25-60 Gy). RESULTS: Three- and 5-year overall survival rates of all patients were 49.5% and 31.3%, respectively. Stratified by symptom sign, 3-year overall survival rate of symptom positive was 27.6% and those of the negative was 56.1% (p = 0.018). Three-year overall survival rates of the total dose > or =51 Gy and that of < or =50 Gy were 58.0% and 42.8%, respectively (p = 0.07). As for morbidity, no patients received Grade 3 or greater late toxicity. CONCLUSIONS: The current study suggested that radiation therapy for isolated para-aortic lymph node recurrence in uterine cervical carcinoma could have a significant impact on survival. PMID- 17126207 TI - Experience in fractionated stereotactic body radiation therapy boost for newly diagnosed nasopharyngeal carcinoma. AB - PURPOSE: Radiotherapy is the most effective treatment for nasopharyngeal carcinoma (NPC). The aim of this study is to evaluate the efficacy and toxicity of fractionated stereotactic body radiation therapy (SBRT) boost for NPC. METHODS AND MATERIALS: Sixty-four patients with newly diagnosed, nonmetastatic NPC were treated with conventional radiotherapy 64.8-68.4 Gy followed by fractionated SBRT boost 12-15 Gy between January 2002 and July 2004. Most patients (72%) presented with Stage III-IV disease. Fifty-two patients also received cisplatin-based concurrent (38) or neoadjuvant (14) chemotherapy. The major endpoints were local control, overall survival, and complications. RESULTS: All patients finished the planned dose of radiotherapy. After a median follow-up of 31 months (range, 22 54), 15 patients developed tumor recurrences--3 in the nasopharynx, 4 in the neck, 5 in distant sites, 1 in both nasopharynx and neck, 2 in the neck and a distant site. The 3-year actuarial rate of local control was 93.1%, regional control 91.4%, freedom from distant metastasis 90.3%, and overall survival 84.9%, respectively. There were no Grade 4 acute or chronic radiation-related complications. CONCLUSIONS: Fractionated SBRT boost for NPC is technically feasible and provides good local control without any severe complications. PMID- 17126208 TI - Radiation tolerance of the rat spinal cord after 6 and 18 fractions of photons and carbon ions: experimental results and clinical implications. AB - PURPOSE: The tolerance of the rat spinal cord to photon and carbon ion irradiations was investigated to determine the relative biologic effectiveness (RBE) of carbon ions ((12)C) in the plateau region and in a 1 cm spread-out Bragg peak. METHODS AND MATERIALS: The cranial part of the cervical and thoracic spinal cord of 336 rats was irradiated with 6 or 18 fractions (Fx) of photons or (12)C ions, respectively. Animals were followed up for 300 days for the onset of paresis grade II and dose-response curves were calculated. RESULTS: The D(50) values (dose at 50% complication probability) were 42.9 +/- 0.5 Gy, 62.2 +/- 0.9 Gy (6 and 18 Fx, (12)C-plateau) and 19.2 +/- 0.2 Gy, 17.6 +/- 0.2 Gy (6 and 18 Fx (12)C-peak), respectively. For photons, the D(50)-values were 57.0 +/- 0.7 Gy for 6 and 88.6 +/- 0.7 Gy for 18 Fx. The corresponding RBE-values were 1.33 +/- 0.02, 1.42 +/- 0.02 (6 and 18 Fx, (12)C-plateau) and 2.97 +/- 0.05, 5.04 +/- 0.08 (6 and 18 Fx (12)C-peak), respectively. Including data of a previously performed experiment for 1 and 2 Fx (1) the parameter alpha/beta of the LQ-model was found to be 2.8 +/- 0.4 Gy, 2.1 +/- 0.4 Gy and 37.0 +/- 5.3 Gy for photon-, (12)C plateau- and (12)C-peak irradiations, respectively. CONCLUSIONS: Carbon ion irradiations of the spinal cord are significantly more effective in the peak than in the plateau region. The alpha/beta-values indicate a significant fractionation effect only for the plateau irradiations. In the Bragg-peak, the applied RBE model correctly describes the main features although it generally underestimates the RBE by 25%. In the plateau region, maximum deviations of up to 20% were found. The acquired data contribute significantly to the validation of the applied RBE-model. PMID- 17126209 TI - Relationship between DNA double-strand break rejoining and cell survival after exposure to ionizing radiation in human fibroblast strains with differing ATM/p53 status: implications for evaluation of clinical radiosensitivity. AB - PURPOSE: To better understand the impact of defects in the DNA damage surveillance network on the various cell-based assays used for the prediction of patient radiosensitivity. METHODS AND MATERIALS: We examined noncancerous human fibroblast strains from individuals with ataxia telangiectasia (ataxia telangiectasia mutated [ATM] deficient) or Li-Fraumeni syndrome (p53 deficient) using the neutral comet, H2AX phosphorylation, and clonogenic survival assays. RESULTS: Using the comet assay, we found that, compared with normal fibroblasts, cells lacking either ATM or p53 function exhibited a reduced rate of double strand break (DSB) rejoining early (< or =4 h) after exposure to 8 Gy of gamma radiation and also exhibited high levels of unrejoined DSBs later after irradiation. ATM-deficient and p53-deficient fibroblasts also exhibited abnormally increased levels of phosphorylated H2AX (gamma-H2AX) at later intervals after irradiation. In the clonogenic assay, ATM-deficient cells exhibited marked radiosensitivity and p53-deficient cells had varying degrees of radioresistance compared with normal fibroblasts. CONCLUSION: Regardless of whether ataxia telangiectasia and Li-Fraumeni syndrome fibroblasts are DSB-repair deficient per se, it is apparent that p53 and ATM defects greatly influence the cellular phenotype as evidenced by the neutral comet and gamma-H2AX assays. Our data suggest that the gamma-H2AX levels observed at later intervals after irradiation may represent a reliable measure of the overall DSB rejoining capabilities of human fibroblasts. However, it appears that using this parameter as a predictor of radiosensitivity without knowledge of the cells' p53 status could lead to incorrect conclusions. PMID- 17126210 TI - The potential of transferrin-pendant-type polyethyleneglycol liposomes encapsulating decahydrodecaborate-(10)B (GB-10) as (10)B-carriers for boron neutron capture therapy. AB - PURPOSE: To evaluate GB-10-encapsulating transferrin (TF)-pendant-type polyethyleneglycol (PEG) liposomes as tumor-targeting (10)B-carriers for boron neutron capture therapy. METHODS AND MATERIALS: A free mercaptoundecahydrododecaborate-(10)B (BSH) or decahydrodecaborate-(10)B (GB-10) solution, bare liposomes, PEG liposomes, or TF-PEG liposomes were injected into SCC VII tumor-bearing mice, and (10)B concentrations in the tumors and normal tissues were measured by gamma-ray spectrometry. Meanwhile, tumor-bearing mice were continuously given 5-bromo-2'-deoxyuridine (BrdU) to label all intratumor proliferating cells, then injected with these (10)B-carriers containing BSH or GB 10 in the same manner. Right after thermal neutron irradiation, the response of quiescent (Q) cells was assessed in terms of the micronucleus frequency using immunofluorescence staining for BrdU. The frequency in the total tumor cells was determined from the BrdU nontreated tumors. RESULTS: Transferrin-PEG liposomes showed a prolonged retention in blood circulation, low uptake by reticuloendothelial system, and the most enhanced accumulation of (10)B in solid tumors. In general, the enhancing effects were significantly greater in total cells than Q cells. In both cells, the enhancing effects of GB-10-containing (10)B-carriers were significantly greater than BSH-containing (10)B-carriers, whether loaded in free solution or liposomes. In both cells, whether BSH or GB-10 was employed, the greatest enhancing effect was observed with TF-PEG liposomes followed in decreasing order by PEG liposomes, bare liposomes, and free BSH or GB 10 solution. In Q cells, the decrease was remarkable between PEG and bare liposomes. CONCLUSIONS: In terms of biodistribution characteristics and tumor cell-killing effect as a whole, including Q cells, GB-10 TF-PEG liposomes were regarded as promising (10)B-carriers. PMID- 17126211 TI - Risk-adaptive optimization: selective boosting of high-risk tumor subvolumes. AB - BACKGROUND AND PURPOSE: A tumor subvolume-based, risk-adaptive optimization strategy is presented. METHODS AND MATERIALS: Risk-adaptive optimization employs a biologic objective function instead of an objective function based on physical dose constraints. Using this biologic objective function, tumor control probability (TCP) is maximized for different tumor risk regions while at the same time minimizing normal tissue complication probability (NTCP) for organs at risk. The feasibility of risk-adaptive optimization was investigated for a variety of tumor subvolume geometries, risk-levels, and slopes of the TCP curve. Furthermore, the impact of a correlation parameter, delta, between TCP and NTCP on risk-adaptive optimization was investigated. RESULTS: Employing risk-adaptive optimization, it is possible in a prostate cancer model to increase the equivalent uniform dose (EUD) by up to 35.4 Gy in tumor subvolumes having the highest risk classification without increasing predicted normal tissue complications in organs at risk. For all tumor subvolume geometries investigated, we found that the EUD to high-risk tumor subvolumes could be increased significantly without increasing normal tissue complications above those expected from a treatment plan aiming for uniform dose coverage of the planning target volume. We furthermore found that the tumor subvolume with the highest risk classification had the largest influence on the design of the risk-adaptive dose distribution. The parameter delta had little effect on risk-adaptive optimization. However, the clinical parameters D(50) and gamma(50) that represent the risk classification of tumor subvolumes had the largest impact on risk adaptive optimization. CONCLUSIONS: On the whole, risk-adaptive optimization yields heterogeneous dose distributions that match the risk level distribution of different subvolumes within the tumor volume. PMID- 17126212 TI - A methodology for using SPECT to reduce intensity-modulated radiation therapy (IMRT) dose to functioning lung. AB - PURPOSE: Single photon emission computed tomography (SPECT) provides a map of the spatial distribution of lung perfusion. Thus, SPECT guidance can be used to divert dose away from higher-functioning lung, potentially reducing lung toxicity. We present a methodology for achieving this aim and test it in intensity-modulated radiotherapy (IMRT) treatment-planning. METHODS AND MATERIALS: IMRT treatment plans were generated with and without SPECT guidance and compared for 5 patients. Healthy lung was segmented into four regions on the basis of SPECT intensity in the SPECT plan. Dose was sequentially allowed to the target via regions of increasing SPECT intensity. This process results in reduction of dose to functional lung, reflected in the dose-function histogram (DFH). The plans were compared using DFHs and F(20)/F(30) values (F(x) is the functional lung receiving dose above x Gy). RESULTS: In all cases, the SPECT guided plan produced a more favorable DFH compared with the non-SPECT-guided plan. Additionally, the F(20) and F(30) values were reduced for all patients by an average of 13.6% +/- 5.2% and 10.5% +/- 5.8%, respectively. In all patients, DFHs of the two highest-functioning SPECT regions were reduced, whereas DFHs of the two lower-functioning regions were increased, illustrating the dose "give take" between SPECT regions during redistribution. CONCLUSIONS: SPECT-guided IMRT shows potential for reducing the dose delivered to highly functional lung regions. This dose reduction could reduce the number of high-grade pneumonitis cases that develop after radiation treatment and improve patient quality of life. PMID- 17126213 TI - Repositioning accuracy of two different mask systems-3D revisited: comparison using true 3D/3D matching with cone-beam CT. AB - PURPOSE: The repositioning accuracy of mask-based fixation systems has been assessed with two-dimensional/two-dimensional or two-dimensional/three dimensional (3D) matching. We analyzed the accuracy of commercially available head mask systems, using true 3D/3D matching, with X-ray volume imaging and cone beam CT. METHODS AND MATERIALS: Twenty-one patients receiving radiotherapy (intracranial/head-and-neck tumors) were evaluated (14 patients with rigid and 7 with thermoplastic masks). X-ray volume imaging was analyzed online and offline separately for the skull and neck regions. Translation/rotation errors of the target isocenter were analyzed. Four patients were treated to neck sites. For these patients, repositioning was aided by additional body tattoos. A separate analysis of the setup error on the basis of the registration of the cervical vertebra was performed. The residual error after correction and intrafractional motility were calculated. RESULTS: The mean length of the displacement vector for rigid masks was 0.312 +/- 0.152 cm (intracranial) and 0.586 +/- 0.294 cm (neck). For the thermoplastic masks, the value was 0.472 +/- 0.174 cm (intracranial) and 0.726 +/- 0.445 cm (neck). Rigid masks with body tattoos had a displacement vector length in the neck region of 0.35 +/- 0.197 cm. The intracranial residual error and intrafractional motility after X-ray volume imaging correction for rigid masks was 0.188 +/- 0.074 cm, and was 0.134 +/- 0.14 cm for thermoplastic masks. CONCLUSIONS: The results of our study have demonstrated that rigid masks have a high intracranial repositioning accuracy per se. Given the small residual error and intrafractional movement, thermoplastic masks may also be used for high precision treatments when combined with cone-beam CT. The neck region repositioning accuracy was worse than the intracranial accuracy in both cases. However, body tattoos and image guidance improved the accuracy. Finally, the combination of both mask systems with 3D image guidance has the potential to replace therapy simulation and intracranial stereotaxy. PMID- 17126214 TI - Dosimetric validation of the MCNPX Monte Carlo simulation for radiobiologic studies of megavoltage grid radiotherapy. AB - PURPOSE: To validate the MCNPX Monte Carlo simulation for radiobiologic studies of megavoltage grid radiotherapy. METHODS AND MATERIALS: EDR2 films, a scanning water phantom with microionization chamber and MCNPX Monte Carlo code, were used to study the dosimetric characteristics of a commercially available megavoltage grid therapy collimator. The measured dose profiles, ratios between maximum and minimum doses at 1.5 cm depth, and percentage depth dose curve were compared with those obtained in the simulations. The simulated two-dimensional dose profile and the linear-quadratic formalism of cell survival were used to calculate survival statistics of tumor and normal cells for the treatment of melanoma with a list of doses of the fractionated grid therapy. RESULTS: A good agreement between the simulated and measured dose data was found. The therapeutic ratio based on normal cell survival has been defined and calculated for treating both the acute and late responding melanoma tumors. The grid therapy in this study was found to be advantageous for treating the acutely responding tumors, but not for late responding tumors. CONCLUSIONS: Monte Carlo technique was demonstrated to be able to provide the dosimetric characteristics for grid therapy. The therapeutic ratio was dependent not only on the single alpha/beta value, but also on the individual alpha and beta values. Acutely responding tumors and radiosensitive normal tissues are more suitable for using the grid therapy. PMID- 17126215 TI - Mapping of nodal disease based on vascular rather than bony anatomy for prostate cancer. PMID- 17126216 TI - In regards to Hall: intensity-modulated radiation therapy, protons, and the risk of second cancers (Int J Radiat Oncol Biol Phys 2006;65:1-7). PMID- 17126218 TI - Neutron dose in scattered and scanned proton beams: in regard to Eric J. Hall (Int J Radiat Oncol Biol Phys 2006;65:1-7). PMID- 17126219 TI - Neutron dose in proton radiation therapy: in regard to Eric J. Hall (Int J Radiat Oncol Biol Phys 2006;65:1-7). PMID- 17126220 TI - Fatal pneumonitis associated with intensity-modulated radiation therapy for mesothelioma: in regard to Allen et al. (Int J Radiat Oncol Biol Phys 2006;65:640 645). PMID- 17126223 TI - Postoperative radiation therapy for prophylaxis of heterotopic ossification of the hip: in regard to Balboni et al. (Int J Radiat Oncol Biol Phys 2006;65:1289 1299). PMID- 17126225 TI - In regard to D'Amico et al.: predictors of mortality after prostate-specific antigen failure (Int J Radiat Oncol Biol Phys 2006;65:656-660). PMID- 17126227 TI - Alternative explanations for T-cell response to in-situ gene therapy for prostate cancer: in reply to Fujita et al. (Int J Radiat Oncol Biol Phys 2006;65:84-90). PMID- 17126229 TI - Cubital tunnel syndrome caused by uremic tumoral calcinosis. PMID- 17126230 TI - Unusual anterior radial head dislocation associated with transposed biceps tendon: a case report. PMID- 17126231 TI - Dislocation of the long head of the biceps tendon with intact subscapularis and supraspinatus tendons. PMID- 17126232 TI - Periprosthetic ulnar fracture after loosening of total elbow arthroplasty treated by two-stage implant revision: a case report. PMID- 17126233 TI - Exposed shoulder hemiarthroplasty as a result of anterosuperior escape: a case report. PMID- 17126234 TI - Arthroscopic osteochondral autograft transfer in the treatment of an osteochondral defect of the humeral head: report of one case. PMID- 17126235 TI - A novel surgical approach to post-traumatic elbow stiffness: case report and operative technique. PMID- 17126236 TI - Ewing's sarcoma of the rotator cuff tendon: a case report. PMID- 17126237 TI - Distant migration of a bioabsorbable implant in the shoulder. PMID- 17126238 TI - Osteoid osteoma of the radial tuberosity presenting as lateral elbow pain. PMID- 17126239 TI - Radial and ulnar neuropathy due to pigmented villonodular synovitis of the elbow. PMID- 17126240 TI - The International Board of Shoulder & Elbow Surgery (established 1992). PMID- 17126241 TI - Rotator cuff repair: the effect of double-row fixation on three-dimensional repair site. AB - There is a high rate of recurrent and residual tears after rotator cuff repair surgery. Recent cadaveric studies have provided surgeons with new knowledge about the anatomy of the supraspinatus tendon insertion. Traditional repair techniques fail to reproduce the area of the supraspinatus insertion, or footprint, on the greater tuberosity anatomically. Double-row suture anchor (DRSA) fixation is a new technique that has been developed to restore the supraspinatus footprint better. In this study, 3-dimensional mapping was used to determine the area of the footprint recreated with 3 different repair methods: a transosseous simple suture technique, fixation with a single row of suture anchors, and DRSA fixation. The DRSA fixation technique consistently reproduced 100% of the original supraspinatus footprint, whereas the single-row suture anchor fixation and transosseous simple suture techniques reproduced only 46% and 71% of the insertion site, respectively. Therefore, the footprint area of the DRSA fixation technique was significantly larger (P < .05) than that of the other 2 techniques. Furthermore, double-row fixation may provide a tendon-bone interface better suited for biologic healing and restoring normal anatomy. PMID- 17126242 TI - Tabletop relocation test: a new clinical test for posterolateral rotatory instability of the elbow. AB - We describe a new clinical test for the assessment of posterolateral rotatory instability. This has been assessed in 8 patients who have been diagnosed with posterolateral rotatory instability. Of these, 6 have undergone surgical reconstruction of the lateral ulnar collateral ligament, and the clinical test has subsequently been negative with resolution of their symptoms. PMID- 17126243 TI - Open surgical treatment of post-traumatic elbow contractures in adolescent patients. AB - The results of surgical treatment of post-traumatic elbow contractures in adolescence have been conflicting in the literature. Twelve adolescent patients (mean age 16.7 years, range 13-21) that had open release of post-traumatic elbow contractures were followed-up for a mean of 18.9 months (range 10-42 months). All releases were performed through a lateral approach (sparing the lateral ulnar collateral ligament) with anterior joint release (in twelve) supplemented by posterior release (in four patients). An additional medial approach was used in three patients. In three patients the radial head was excised. A mean gain of 54 degrees in the flexion-extension arc was observed at final follow-up and all patients achieved a functional ROM of at least 100 degrees. The patients maintained 93% of the motion that was achieved intraoperatively. No patient lost motion. Open release in adolescent patients with post-traumatic elbow contractures and no intarticular incongruence or erosion, yielded satisfactory results, similar to those achieved in adults. PMID- 17126244 TI - Simulated humeral avulsion of the glenohumeral ligaments: a new instability model. AB - Humeral avulsion of the glenohumeral ligaments (HAGL) is an infrequent cause of shoulder instability. Experimental studies on this lesion are rare. This study was undertaken to determine the extent of humeral-based capsuloligamentous damage required for dislocation to occur. In 65 fresh cadaver shoulders, a humeral-sided ligamentous cutting sequence was done. After each step, degree of sulcus, translation, and instability were evaluated with an electromagnetic tracking device. There was a high degree of correlation between the amount of cut done and the resulting degree of instability. The order of the ligamentous cuts had no significant influence. For a dislocation to occur at least 3 zones had to be cut. Simulated HAGL can be used as a model for shoulder instability, although further experiments are needed to validate this model fully. Extensive capsuloligamentous lesions on the humeral side seem to be required before dislocation can occur. This may be a factor explaining the relative paucity of HAGL in clinical series. PMID- 17126245 TI - The biomechanical role of the subscapularis in latissimus dorsi transfer for the treatment of irreparable rotator cuff tears. AB - Inferior functional results of latissimus tendon transfer for the treatment of irreparable rotator cuff tears have been reported in the presence of a subscapularis tendon tear. A biomechanical or experimental explanation for the necessity of an intact subscapularis is unavailable. It was, therefore, the purpose of this investigation to study the biomechanical role of the subscapularis in the treatment of a posterosuperior rotator cuff tear with latissimus dorsi transfer. A biomechanical cadaveric model was developed to reproduce glenohumeral motion patterns created by loading of the transferred latissimus dorsi tendon with and without simultaneous action of the subscapularis muscle. Significant differences could be demonstrated not only for translation but also for rotation of the humeral head depending on subscapularis action. In the neutral and in the abducted/externally rotated position of the arm, anterior translation and dislocation of the joint were encountered without subscapularis action. Our results provide evidence that motion patterns of the humeral head are significantly altered in the absence of the stabilizing effect of the subscapularis muscle. The inferior functional results of latissimus dorsi transfer in the presence of subscapularis dysfunction are explained by the loss of centering of the humeral head upon abduction and elevation if subscapularis function is deficient. PMID- 17126246 TI - Non-mineralized fibrocartilage shows the lowest elastic modulus in the rabbit supraspinatus tendon insertion: measurement with scanning acoustic microscopy. AB - The acoustic properties of rabbit supraspinatus tendon insertions were measured by scanning acoustic microscopy. After cutting parallel to the supraspinatus tendon fibers, specimens were fixed with 10% neutralized formalin, embedded in paraffin, and sectioned. Both the sound speed and the attenuation constant were measured at the insertion site. The 2-dimensional distribution of the sound speed and that of the attenuation constant were displayed with color-coded scales. The acoustic properties reflected both the histologic architecture and the collagen type. In the tendon proper and the non-mineralized fibrocartilage, the sound speed and attenuation constant gradually decreased as the predominant collagen type changed from I to II. In the mineralized fibrocartilage, they increased markedly with the mineralization of the fibrocartilaginous tissue. These results indicate that the non-mineralized fibrocartilage shows the lowest elastic modulus among 4 zones at the insertion site, which could be interpreted as an adaptation to various types of biomechanical stress. PMID- 17126247 TI - Effect of rotator interval closure on glenohumeral stability and motion: a cadaveric study. AB - The effect of rotator interval closure, which is performed as an adjunct to arthroscopic stabilization of the shoulder, has not been clarified. Fourteen fresh-frozen cadaveric shoulders were used. The position of the humeral head was measured using an electromagnetic tracking device with the capsule intact, sectioned, and imbricated between the superior glenohumeral ligament and the subscapularis tendon (SGHL/SSC closure) or between the superior and middle glenohumeral ligaments (SGHL/MGHL closure). The direction of translational loads (10, 20, and 30 N) and arm positions were (1) anterior, posterior, and inferior loads in adduction; (2) anterior load in abduction/external rotation in the scapular plane; and (3) anterior load in abduction/external rotation in the coronal plane. The range of motion was measured using a goniometer under a constant force. Both methods reduced anterior translation in adduction. Only SGHL/MGHL closure reduced anterior translation in abduction/external rotation in the scapular plane and posterior translation in adduction. Both methods reduced the range of external rotation and horizontal abduction. Rotator interval closure is expected to reduce remnant anterior/posterior instability and thereby improve the clinical outcomes of arthroscopic stabilization procedures. PMID- 17126248 TI - Sialoblastoma: a clinicopathologic and immunohistochemical study of 7 cases. AB - Sialoblastoma is a rare congenital or perinatal salivary tumor that varies in histologic features and biologic potential. Seven cases from the files of the Armed Forces Institute of Pathology are presented. These tumors occurred in 4 males and 3 females with ages ranging from prenatal to 6 months at the time of discovery. Five lesions originated from the parotid gland; 2 lesions were from the submandibular gland. All lesions presented as nodular to multinodular swellings and ranged in size from 2.0 to 7.0 cm. The principal sign or symptom was rapid growth. Two histologic patterns with differing behavior predominated: (1) a favorable pattern had semiencapsulation of cytologically benign basaloid tumor cells with intervening stroma; and (2) an unfavorable histology of anaplastic basaloid tumor cells, minimal stroma, and broad pushing to infiltrative periphery. Four and three tumors had favorable and unfavorable growth patterns, respectively. One unfavorable lesion had vascular invasion, and another demonstrated perineural invasion. All 3 tumors with unfavorable histology recurred. Tumor cells in 3 cases were immunohistochemically reactive for keratin, S-100, smooth muscle actin, and calponin to varying degrees. All 3 tumors were reactive for p63. alpha-Fetoprotein was expressed in 2 unfavorable tumors. Ki67 was expressed at 3% in a favorable tumor and 40% and 80% in the 2 unfavorable lesions. Treatment involved surgical excision. One patient received adjuvant chemotherapy. Two sialoblastomas resulted in recurrences within a year and another developed a recurrence after 4 years. One sialoblastoma developed lung metastasis within 1 month of the original biopsy. Although a clinical correlation is suggested by a favorable/unfavorable histologic grading system the biologic behavior is nonetheless considered unpredictable. PMID- 17126249 TI - The status of cyclooxygenase-2 expression in ductal carcinoma in situ lesions and invasive breast cancer correlates to cyclooxygenase-2 expression in normal breast tissue. AB - OBJECTIVES: There is a paucity of data on cyclooxygenase (COX)-2 expression in normal breast tissue and on the changes in COX-2 expression from normal tissue via ductal carcinoma in situ (DCIS) lesions to invasive cancer. The aim of this study, therefore, was to investigate COX-2 protein expression in normal breast tissue, DCIS, and invasive breast cancer in samples from the same patients. METHODS: In 39 patients, we investigated and compared COX-2 expression in paired samples of invasive cancer and normal adjacent breast epithelium by immunohistochemistry with a monoclonal COX-2 antibody. Furthermore, in 29 of these cases, we also analyzed a concomitant DCIS lesion. RESULTS: Patients without COX-2 expression in normal breast tissue also do not express COX-2 in invasive breast cancer and in DCIS lesions, respectively. Conversely, COX-2 expression in normal breast tissue was an indicator for COX-2 expression in the paired breast tumors. There was no significant correlation between COX-2 expression and pathologic tumor stage, nodal status, hormone receptor status, tumor size, grading, and lymphovascular space involvement. CONCLUSIONS: This is the largest study to date investigating COX-2 in paired samples of breast tumors and normal adjacent breast tissue. Our data are consistent with the hypothesis that COX-2 overexpression is an early event in breast carcinogenesis. PMID- 17126250 TI - Flat elevated lesions of the colon and rectum: a spectrum of neoplastic and nonneoplastic entities. AB - The aim of this prospective study is to establish the frequency and the type (neoplastic and nonneoplastic) lesions defined endoscopically as flat elevated lesion (FEL) in the colon and rectum, as well as to compare flat adenomas (FAs) to polypoid lesions of the same size with morphometric and immunohistochemical analysis. One hundred nineteen patients were studied through fibrocolonoscopy with chromoscopy (indigo carmine spray). All detected lesions (total of 195) were removed, and FELs measuring 10 mm or smaller were also selected. Using histopathologic criteria, they were divided in neoplastic (adenomas and carcinomas) and nonneoplastic ones. In neoplastic lesions, the following parameters were evaluated to compare FAs with polypoid lesions: morphometric studies with Index of Structural Atypia (ISA) and Stratification Index (SI), evaluation of cellular proliferation with label index of Ki-67, and expression of p53 protein. Of 195 lesions resected, only 33 (17%) met the endoscopic requirements for FELs. Twelve (36.4%) were neoplastic and 21 (63.6%) considered nonneoplastic. Among the FAs, there were a percentage of high-grade (severe dysplasia) significantly more frequent than observed in polypoid lesions (16.7% vs 2.6%). In addition, the SI, Ki-67 label index and p53 positivity were significantly higher in FAs. The ISA also reached significant differences between both groups of adenomas. Non-neoplastic FELs included different entities such as hyperplasic polyps, focuses of colitis, normal mucosa, and scars. The endoscopic elements analyzed were shared between nonneoplastic FELs and FAs. A central depression, when air was properly insufflated, considered typical in neoplastic lesions, was frequently observed in nonneoplastic lesions. Following the endoscopic criteria of FELs, nonneoplastic lesions predominated over the adenomatous lesions, demonstrating that FELs and FAs are not homologous terms. The frequency of high-grade dysplasia was significantly more elevated in the adenomatous FELs than in polypoid adenomas. The ISA, SI, p53 expression, and Ki 67 label index were helpful in differentiating adenomatous FELs from polypoid lesions. Flat elevated lesions selected by endoscopic criteria are, in fact, a heterogeneous population of lesions. PMID- 17126251 TI - Pharyngeal teratocarcinosarcoma: review of the literature and report of two cases. AB - Teratocarcinosarcomas are rare malignant neoplasms histologically characterized by the presence of benign and malignant epithelial and mesenchymal elements. They are seen almost exclusively in the sinonasal tract of men. We report two cases of teratocarcinosarcomas involving the posterior pharyngeal wall in a 55-year-old male and 60-year-old men. The tumors consisted of epithelial components including squamous, neuroendocrine, and glandular structures; neuroepithelium, and mesenchymal components with prominent rhabdomyoblastic, osteoblastic and chondroid differentiation. Immunohistochemical studies demonstrated markers characteristic of each component. The tumors were resected, and the patients received postoperative radiation therapy. One patient is alive with recurrent tumor 33 months after treatment and the other died 26 months after radiation therapy with distant metastasis. PMID- 17126252 TI - Desmoplastic fibroma of bone with extensive cartilaginous metaplasia. AB - Desmoplastic fibroma of bone is a rare tumor demonstrating the same histologic and biologic features of its soft tissue counterpart, aggressive fibromatosis. We report the second case of desmoplastic fibroma of bone with extensive chondroid metaplasia. The tumor arose in the left ischium of a 51-year-old male, with extension into adjacent musculature as a pseudoencapsulated mass. The infiltrating growth and quality of the fibrous component are characteristic of desmoplastic fibroma, and in addition, abrupt transitions into bland hyalin cartilage were frequent. Discriminating features of this lesion from other bone tumors capable of biphasic expression of fibrous and chondroid elements are discussed. PMID- 17126253 TI - Hirschsprung's disease in a young adult: report of a case and review of the literature. AB - Hirschsprung's disease (HD) in adults is rare and often undiagnosed or misdiagnosed. We report a case of HD in a 26-year-old woman who had a history of chronic constipation that required laxatives and enemas since early childhood. She developed severe intestinal obstruction and presented to the emergency department with significant abdominal distension. A computed tomographic scan confirmed significant fecal loading of the entire colon and rectum. An anal manometry revealed lack of normal rectoanal inhibitory reflex. A rectal biopsy showed hypoganglionic anorectum, suspicious for HD. Because of the severe fecal retention that was refractory to conservative management, total proctocolectomy with ileal pouch-anal anastomosis was performed. The entire colon showed massive dilatation and marked wall thickening. Histologic examination showed absence of ganglion cells in submucosal (Meissner's) and myenteric (Auerbach's) plexuses in the distal rectum. A diagnosis of adult HD was made. Her postoperative course was uneventful with complete resolution of the symptoms. Hirschsprung's disease should be considered in adults who have long-standing and refractory constipation. PMID- 17126254 TI - Diffuse intra-abdominal clear cell myomelanocytic tumor: report of an unusual presentation of "PEComatosis" simulating peritoneal mesothelioma. AB - We report a case of diffuse myomelanocytic tumor of the peritoneum that simulates, clinically and instrumentally, a malignant mesothelioma. The patient was a 70-year-old woman with a history of ancient hysterectomy for fibroids, who presented with abdominal discomfort. Exploratory laparotomy revealed diffuse encasing of the peritonealized organs by a thin, fleshy, gray-white tissue rind. Scattered tumor masses were present as well. A dominant lesion measuring 6 cm in larger size was resected from the pelvis. Histological examination revealed a tumor composed of epithelioid and spindle cells, exhibiting either a clear or slightly eosinophilic cytoplasm and a mild to moderate nuclear pleomorphism. Focal areas of necrosis could be documented. Immunohistochemically, tumor cells were positive for HMB45, melan-A, and smooth muscle actin, but negative for other antibodies, including epithelial markers, desmin, and S100 protein. We believe that this case represents an example of myomelanocytic tumor of uncertain biologic potential, a member of the recently devised perivascular epithelioid cell tumors (PEComa), with an unusual presentation simulating a diffuse mesothelial neoplasm. The origin of this particular lesion is briefly discussed in light of the recent literature published on the subject. PMID- 17126255 TI - Mediastinal follicular dendritic cell sarcoma involving bone marrow: a case report and review of the literature. AB - We report a rare case of mediastinal follicular dendritic cell (FDC) sarcoma involving the bone marrow. The patient, a 46-year-old woman, had a clinically aggressive tumor in the anterior mediastinum that was initially diagnosed as a diffuse B-cell lymphoma. She received chemotherapy but showed no significant improvement. One year later, the patient presented at our institution with pelvic bone metastases. Biopsy specimens of the sacrum lesion and bone marrow were obtained. The diagnosis of FDC sarcoma was made based on histological examination and immunohistochemical findings, including strong positive staining of tumor cells for CD21, CD23, clusterin, and epidermal growth factor receptor (EGFR) and negative staining for CD20, CD30, CD45, CD1a, S-100, vimentin, and keratin cocktail. Histological examination and immunohistochemical studies of a previous biopsy of the mediastinal mass confirmed the diagnosis of mediastinal FDC sarcoma. The patient was treated with an appropriate chemotherapy regimen; 1 month later, follow-up bone marrow biopsy revealed no tumor cells. Although FDC sarcoma is considered a low-grade tumor, the tumor in the present case not only developed at an unusual location with bone metastasis but also involved bone marrow. To our knowledge, this is the first such case ever reported. This case also highlights the utility of EGFR as an immunohistochemical marker of dendritic cell tumors that could be used as a diagnostic tool and guide for choosing appropriate chemotherapy regimens. PMID- 17126256 TI - Ewing sarcoma/primitive neuroectodermal tumor of the kidney: a case report. Diagnosed by immunohistochemistry and molecular analysis. AB - BACKGROUND: Ewing sarcoma/primitive neuroectodermal tumor (EWS/PNET) of the kidney is a rare and aggressive tumor. It has a rapid clinical progression with early metastasis and death. Few cases with documented t(11;22) have been reported in the literature. CASE PRESENTATION: We report a case of EWS/PNET of the kidney in a 26-year-old woman with widespread metastasis at initial presentation. The tumor cells showed strong expression for CD99 and FLI-1 monoclonal antibodies and polyclonal antibodies and were negative for WT1 and numerous other markers. The diagnosis was subsequently confirmed by demonstrating t(11;22)(q24;q12) using cytogenetic karyotyping and fluorescence in situ hybridization. CONCLUSIONS: Due to the different prognosis and management between EWS/PNET and other primary renal neoplasms with similar morphology, a histopathologic diagnosis with extreme accuracy should be made. Cytogenetic analysis is an important supportive tool to immunohistochemistry in making the final diagnosis. PMID- 17126257 TI - Benign hybrid perineurioma-schwannoma in the colon. A case report. AB - A 48-year-old man of Haitian descent presented with progressive constipation, hematochezia, and weight loss. Colonoscopy and computed tomography scan revealed an obstructing colonic mass, causing intussusception and pneumatosis of the descending/upper sigmoid colon and necessitating an emergency left hemicolectomy. Gross examination revealed a 4.9-cm obstructing mass in the sigmoid colon extending through the muscularis propria. Histologically, the lesional cells were bland, spindled, with tapered and often wavy nuclei set in a loose fibromyxoid stroma. Focally, the lesional cells displayed whorling or storiform growth pattern mixed with spindle wavy cells. In many areas, the cells had bipolar cytoplasmic processes. Immunohistochemistry revealed patchy positivity for epithelial membrane antigen, CD34, vimentin, diffuse positivity for S-100, and negativity for CD117, cytokeratin (AE1/AE3), ALK1, desmin, smooth muscle actin, neuronal nuclei antigen (NeuN), and neurofilament protein. The morphology and immunohistochemical findings were consistent with hybrid perineurioma-schwannoma. Electron microscopic examination revealed the characteristic features of perineuriomal and schwannomatous differentiation. Based upon the histopathology, immunophenotype, and ultrastructure, this tumor was classified as a benign hybrid perineurioma-schwannoma, a counterpart to the tumor described in the soft tissue. This is the first case report of hybrid perineurioma-schwannoma in the colon. PMID- 17126258 TI - Mesothelial cyst of the pancreas. AB - Cystic lesions of the pancreas are increasingly identified because of advances in imaging techniques. The cystic lesions are of different types and are classified as neoplastic, nonneoplastic, and developmental types. Identification of the nature of these cystic lesions is very important because the course and treatment of disparate types of cysts are different. Here, we describe, for the first time, mesothelial cyst involving the pancreas in a 36-year-old man. Distal pancreatectomy showed a 3-cm unilocular cyst containing clear fluid. The cyst was lined by flat to cuboidal epithelium surrounded by fibrous tissue. The lining epithelial cells were positive for vimentin, thrombomodulin, cytokeratin 5/6, and calretinin, thus confirming the mesothelial nature of the cells. PMID- 17126259 TI - Intestinal metastases in a squamous cell carcinoma arising in mature cystic teratoma of the ovary: a case report. AB - Metastatic spread to the intestines by squamous cell carcinoma (SCC) arising in mature cystic teratoma (MCT) or dermoid cyst is a very rare event. A case of a 40 year-old cachectic woman is presented with an MCT of the left ovary found at autopsy. Focal mural thickening of the cyst wall harbored a moderately differentiated SCC with single cell keratinization. Intramural metastases were noted in the jejunum, ileum, colon transversum, and left colonic flexure. In addition, liver metastases were detected. The tumor was staged as FIGO IV. The peritoneum was covered diffusely by fibrinous and suppurative exudate. In this unusual case, metastasizing SCC of the ovary arising in an MCT led to suppurative peritonitis and consequently to death of the patient. Follow-up procedures regarding intestinal spread could be useful in patients with SCC in MCT. PMID- 17126260 TI - The Toker tumor: spectrum of morphologic features in primary neuroendocrine carcinomas of the skin (Merkel cell carcinoma). AB - Primary neuroendocrine carcinoma of the skin is a relatively rare tumor that was first described by Cyril Toker in 1972. Since the seminal paper by Toker based on simple morphologic observations and detailed clinical correlation, our understanding of the clinical, morphological, and biological attributes of these lesions has grown exponentially with their increased awareness by pathologists and clinicians as well as with the many contributions of modern diagnostic techniques. The present review focuses principally on the various morphologic appearance that these tumors are able to adopt, the role of modern special techniques for diagnosis, and the conditions that need to be considered in their differential diagnosis. PMID- 17126261 TI - End-to-side nerve coaptation: a qualitative and quantitative assessment in the primate. AB - There are several reasons why end-to-side nerve coaptation has not been widely adopted clinically. Among these are the putative damage inflicted on the donor nerve and the variable quality of the regeneration in the recipient nerve. So far experiments on end-to-side nerve repair have been short term and mostly carried out on rats. This long-term study of end-to-side nerve repair of ulnar to median and median to ulnar nerve was performed using adult nonhuman primates. Eleven nerve repairs were studied at different time points. Eighteen, 22, 33 and 57 months after surgery a qualitative and quantitative analysis of the donor nerve and regenerating nerve revealed variable levels of percentage axonal regeneration compared with matched controls (1.4%-136%). Morphological evidence of donor nerve damage was identified distal to the coaptation site in four of the 11 cases, and in these cases the best axonal regeneration in the corresponding recipient nerves was observed. This donor nerve damage could neither be demonstrated in terms of a decrease in axon counts distal to the coaptation nor as donor target organ denervation. Recipient target organ regeneration like the axonal regeneration varied, with evidence of motor regeneration in eight out of 11 cases and sensory regeneration, as measured by percentage innervation density compared with matched controls, varied from 12.5% to 49%. Results from the present study demonstrate that the end-to-side coaptation technique in the nonhuman primate does not give predictable results. In general the motor recovery appeared better than the sensory and in those cases where donor nerve damage was observed there was better motor and sensory regeneration overall than in the remaining cases. PMID- 17126262 TI - Precision grip function after free toe transfer in children with hypoplastic digits. AB - Although toe-to-hand transfer has a defined role in the management of congenital hand deformities, it remains unclear how well children integrate the transferred digits into physiological grasping. We analysed fingertip forces in the precision grip of 13 patients when lifting a test object more than three years after free toe transfer for absent or hypoplastic digits. Clinically, most patients showed normal sensibility of transferred digits, but active motion and pinch strength were limited as compared to the normal hand. For the control of fingertip forces, two key features of the normal two-digit opposition grip were seen in all operated hands: adaptation of grip force to object weight and parallel coordination of lift and grip forces. These physiological grasping strategies developed independently of the patients' age at the time of operation, which ranged from one to 13 years. In four patients, we observed increased tangential load forces with the operated hand due to misalignments in the application of fingertips on the grasp surfaces. Such forces lead to increased grip force requirements on both fingers that may overload transferred digits with limited motor function. The need for optimal alignment of the grip axis during toe transfer surgery is emphasised. PMID- 17126263 TI - Melanoma of thumb: retrospective study for amputation levels, surgical margin and reconstruction. AB - For the treatment of melanoma of thumb, the evidence about amputation level (IP joint or MP joint) and cutaneous excision margin has not been fully presented. And reconstructions for lost thumbs need to be balanced against functional and aesthetic requirements of individual patients and overall prognosis. We report 15 patients, who underwent primary excision and reconstruction for thumb melanoma between 1986 and 2004 at Department of Plastic and Reconstructive Surgery, University of Hokkaido at Sapporo, Graduate School of Medicine. The patients were reviewed to evaluate the prognostic significance of variables including age and sex of the patient, tumour thickness, staging, level of amputation, and cutaneous excision margin. Our 15 cases were divided into two groups according to amputation level (MP or IP) and cutaneous excision margin (<40 mm or >or =40 mm) and prognosis was analysed statistically. The stage and thickness were identified as prognostic factors for disease-free survival; however, IP amputation and 10-30 mm margin did not compromise disease-free survival. Amputated thumb was reconstructed by pollicization, free toe to thumb transfer, reverse forearm flap, local flap, and skin graft. No major complication of reconstructed thumb occurred. Seven of eight cases of MP amputation were reconstructed by pollicization. On the other hand, five cases of IP amputation were reconstructed by volar skin flap in two cases, reverse forearm flap in two cases, and trimmed first toe transfer in one case. Although a small number of cases, our study is the first attempt focussing on melanoma of thumb, and evaluating amputation level and cutaneous margin separately. The reconstructive algorithm for the amputated thumb is proposed and has various tools including free toe to thumb transfer. PMID- 17126264 TI - Incidence, mortality and survival in cutaneous melanoma. AB - Cutaneous melanoma remains a challenge despite increased levels of awareness, education and targeted health policies. Worldwide incidence rates for cutaneous melanoma have risen faster than those for any other malignancy in Caucasian populations over the last 30 years. Despite improving survival rates (defined as the ratio of those who survive the disease against incidence) over this period, mortality rates, generally, have continued to climb. Mortality from melanoma is greater than that caused by all other types of skin cancer, especially in men. In Britain the percentage of increase in the male age standardised mortality rate surpassed that of all other malignancies assessed (1993-2002) by Cancer Research UK. A literature-based study was conducted with review of publications identified through Medline and EMBase, 1980-December 2005, databases. We present a review of the current literature on incidence, mortality and survival rates of melanoma including a discussion on the aetiological factors, behaviour modification associated with public education campaigns and recent health policies and the effect these are having on melanoma figures. It is likely that any fall in mortality rates from melanoma in the near future will be secondary to early detection. Changes resulting from primary prevention are unlikely to be noticeable for several decades. PMID- 17126265 TI - Basal cell carcinoma histological clearance margins: an analysis of 1539 conventionally excised tumours. Wider still and deeper? AB - An analysis of peripheral and deep margins of histological clearance around 1539 consecutive basal cell carcinomas excised by conventional surgery showed that 81 lesions (5.3%) were incompletely excised peripherally; 36 lesions (2.3%) were incompletely excised deeply; 13 lesions (0.8%) were incompletely excised peripherally and deeply. Nine hundred and ninety-six lesions (65%) were excised with a peripheral histological clearance margin<5mm (0.1-4.9mm), whereas 1303 lesions (85%) were excised with a deep histological clearance margin<5mm (0.1 4.9mm). Four hundred and eight lesions (27%) had a peripheral histological clearance margin of 5.0-9.9mm, whereas 170 lesions (11%) had a deep histological margin of 5.0-9.9mm. Peripheral histological clearance margins exceeded 10mm in 41 lesions (3%) and deep histological margins exceeded 10mm in 17 lesions (1%). Thus 30% of peripheral histological margins were 5mm or more but only 12% of deep histological margins were 5mm or more. Despite a relative sparing of deep tissue, incomplete excision in depth affected only 36 lesions compared with 81 incomplete peripheral excisions. Peripheral histological clearance was <5mm (0.1-4.9mm) for 55% of temple lesions, 50% of scalp lesions and 43% for limb lesions. In the cosmetically sensitive areas of peri-orbital region, nose, cheek, lip, neck and chin more than 70% of lesions were excised with a peripheral histological margin<5mm. This study of conventional surgical excision of basal cell carcinomas with an incomplete excision rate of 8% has shown that 65% of lesions were excised with <5mm histological clearance peripherally and 85% with <5mm deep clearance. These figures for 'normal tissue sacrifice' are not excessive when compared with those of 'tissue sparing' Mohs' micrographic surgery in which the operator may take a margin of several millimetres of normal tissue in the initial 'slice', or in the subsequent 'safety margin' beyond the eventual tumour free plane. However, peripheral margins did exceed 5mm in more than 30% of lesions of scalp, temple and forehead, and for these sites where even with loupe magnification the tumour edge could be difficult to define, either frozen section control or Mohs' technique, might with benefit be more often used in order to minimise normal tissue sacrifice. PMID- 17126266 TI - Hypospadias and associated penile anomalies: a histopathological study and a reconstruction of the pathogenesis. AB - Hypospadias is considered to be the result of inadequate fusion of urethral folds and, possibly, of canalization of a glandar epithelial cord during the formation of the spongy urethra. This theory had to be reconsidered because a recent study in normal human embryos has exposed such fusion and invagination as misconceptions. Autopsy specimens of five penises with hypospadias from foetuses and neonates were studied histologically. The findings complemented with data from the literature were correlated to the normal developmental process to reconstruct the pathogenesis of the disorder. Histopathological analysis revealed that the hypospadic orifice was the proximal part of a mucosal delta which revealed the structure of the roof and meatus of a flattened distal urethra. Branches of the raphe bordering the delta and terminating in prominent 'dog ears' had the characteristics of the transient urethral labia (folds). Associated curvature and torsion could be related to structural abnormalities of vascular structures, notably the distal corpus spongiosum, and fasciae predominantly proximal to the hypospadic orifice. Correlation with normal development indicated that hypospadias and associated anomalies are not caused by disturbed fusion or glandar invagination but by maldevelopment of a complex of primordial fascial and vascular tissue proximal to the urethral orifice which normally form the venter side of the penis by disproportionately strong proliferation and make the urethral orifice shift distalward. Insufficient growth may disturb that shift with the degree of deficiency determining the precise position of the urethral orifice, size of the urethral delta and defect of the prepuce. Shortage and/or poor organisation of these tissues explain curvature and, if asymmetrical, torsion, both of which can occur also with minimal urethral deformity or as congenital ventral curvature and torsion without hypospadias. PMID- 17126267 TI - Centralisation of a hypospadias repair service--the Warwickshire experience. AB - An audit of the hypospadias repair service provided in the county of Warwickshire has shown that centralising all referrals to a single surgeon who uses limited, well-practiced techniques, greatly improves outcome. Comparison to previous audit of the service, when provided by a number of surgeons in the region, shows complication rates dropping - fistulae occurrence reducing from 35.8% to 6.7% and urethral stricture rates showing similar improvement from 4.4% to 0%. Overall productivity increased, and referral out of region dropped from 13% to 0%. This provides definitive evidence that restructuring of a service already in place has decreased morbidity and increased efficiency to give results comparable to published data. PMID- 17126268 TI - Reduction of hypertrophic scar via retroviral delivery of a dominant negative TGF beta receptor II. AB - Effective blockade of the pluripotent cytokine transforming growth factor (TGF) beta as a means of cutaneous scar reduction is a strategy with great potential. This desired effect may be achieved through the overexpression of mutant TGF-beta receptors within the wound milieu. Our goal was to examine the effects of dominant negative mutant TGF-beta receptor II (TGFbetaRIIdn) protein expression in a well-established rabbit ear model of hypertrophic scarring. Serial injections of a retroviral construct encoding a truncated TGFbetaRII and the marker green fusion protein (pMSCV-rIIdn-GFP) were performed in 7mm punch wounds at day 10 and day 12 (two-day injection group) or days 8, 10, 12 (three-day injection group) post-wounding. Delivery of an empty vector (pMSCV-GFP) at the same time points served as a negative control. Histomorphometric analysis of wounds harvested at day 28 revealed a modest, though statistically significant reduction (20%, p=0.027) in the scar elevation index (SEI) in two-day treated and a more modest reduction in SEI (12%) in the three-day treated arm compared to null-treated controls. Confocal microscopy confirmed stable, yet variable transfection of the construct in both peri-wound tissue as well as rabbit dermal fibroblasts transfected in vitro. Optimisation of this novel application in retroviral gene therapy could lead to effective anti-scarring strategies. PMID- 17126270 TI - Patient satisfaction following abdominoplasty: an NHS experience. AB - Abdominoplasty is one of the few cosmetic procedures performed in the UK on the National Health Service (NHS). In the current climate of tight resource rationing many primary care trusts (PCTs) are refusing to fund such operations, resulting in a number being carried out in the independent sector. There are very few studies in the literature on patient satisfaction following abdominoplasty, with none from the UK. Our study aimed to look at the patient satisfaction rate following abdominoplasty in the NHS and study further the areas of dissatisfaction. A questionnaire survey was sent to 153 patients who underwent the procedure over a 5-year period. Replies were received from 118 (78%) patients, revealing a 77% satisfaction rate amongst responders. Further analysis of the dissatisfied patients revealed particular dissatisfaction with dog-ears, residual abdominal overhang and issues relating to the scar. The dissatisfied group was exclusively female from across the age spectrum, although analysis showed an increase in dissatisfaction with time, with the highest levels seen at 4-6 years post-abdominoplasty. This study has identified a number of areas of patient dissatisfaction, even in carefully selected patients. PMID- 17126271 TI - The validation of the Depth Measurement Videomicroscope (DMV) as a noninvasive tool for the assessment of capillary vascular malformations. AB - The assessment of capillary vascular malformation (CM) morphology can be performed using videomicroscopy. Previously only the type of capillary pattern could be demonstrated. The Depth Measurement Videomicroscope (DMV) allows both depth and diameter of CM vessels to be measured. The aim of this study was to examine how videomicroscope recordings correlated with biopsy recordings and to investigate pressure-related changes in recordings when using the device. For the first part of the study, 10 patients with CMs resting in a temperature-controlled room were assessed with the DMV. Following this a 3mm punch biopsy of the area was taken. The depth and diameter measurements taken with the DMV were compared to those obtained histologically. For the second part of the study, pressure measurement was used to determine the amount of pressure required on the tip of the DMV to alter the results obtained. Five recordings were taken on the forearm of one volunteer. When the DMV and biopsy measurements are compared using a Bland and Altman Test to determine their relationship there is a close agreement with the diameter measurements and a correction factor of -0.100mm for the depth measurements. The pressure required to alter the skin microcirculation when placing the DMV on the skin surface was found to be 62mmHg. This corresponds closely with other studies of pressure effects on the skin microcirculation and exceeds the pressure used when using the DMV. The DMV thus provides a useful tool for assessing CM capillary structure. PMID- 17126272 TI - Use of a non-contact 3D digitiser to measure the volume of keloid scars: a useful tool for scar assessment. AB - Keloid scars often fail to respond to treatment, so research into new therapeutic regimes is important. However, research is limited by a scarcity of reliable, objective scar assessment tools. The volume of a keloid scar should decrease with successful treatment. This study demonstrates the use of a non-contact 3D digitiser to measure digitally the volume of a keloid scar. The scanner was used to scan 62 keloid scars and one fine-line normal scar. The scan took approximately 9s to complete. The volume was measured using 3D reverse modelling software. A previously validated scar assessment scale was used to score the scars according to their physical parameters. A significant correlation was found between volume and the scar score (Pearson's r=0.627, p<0.001). Linear regression was also statistically significant (p<0.001, R(2)=0.44). Therefore it was possible to predict the scar score from the measured volume. This technique could allow monitoring of a patient on treatment, or comparison of treatments in a research setting. It overcomes previous problems with the measurement of scar volume as it is quantitatively objective and well-tolerated. PMID- 17126273 TI - Transaxillary endoscopic release of restricting bands in congenital muscular torticollis--a novel technique. AB - Congenital muscular torticollis is due to fibrosis of one or both the heads of sternocleidomastoid muscle. This may also involve the platysma, scalene muscles, and the carotid sheath and may be associated with cervical scoliosis. Conventional surgical procedures leave visible scars. Ramirez, who used the posterior part of the traditional face-lift incision, made perhaps the first attempt at concealing scars. Burstein et al. reported a large series of subcutaneous endoscopic release of torticollis through a hairline approach. Sasaki described an endoscopic two-incision, posterior auricular fold and hairline approach. A technique of transaxillary subcutaneous endoscopy for the release of the sternocleidomastoid muscle in congenital muscular torticollis is described here. This procedure provides direct access to the fibrous bands, enables release without risk of damage to the spinal accessory nerve, external jugular vein, or greater auricular nerve, and leaves no visible neck scars. Two cases of congenital muscular torticollis presenting in adulthood were managed successfully by this technique. The fibrotic part of sternocleidomastoid muscle was released and the normal range of head motion was restored. There were no surgical complications encountered and the patients achieved complete pain free range of movement in six weeks. This technique provides direct and quick access, perpendicular to the line of the fibrotic bands, avoids injury to neurovascular structures and does not leave visible neck scars. PMID- 17126274 TI - A novel technique for vacuum assisted closure device application in noncontiguous wounds. PMID- 17126275 TI - Earlobe perforation after prolonged use of a topical corticosteroid. PMID- 17126276 TI - Microvascular anastomosis: is there a role for robotic surgery? PMID- 17126277 TI - An alternative device for illuminated retraction in augmentation mammaplasty. PMID- 17126278 TI - Determining the potential safety benefit of improved lighting in three pedestrian crash scenarios. AB - The influence of light level was determined for three pedestrian crash scenarios associated with three adaptive headlighting solutions-curve lighting, motorway lighting, and cornering light. These results were coupled to corresponding prevalence data for each scenario to derive measures of annual lifesaving potential. For each scenario, the risk associated with light level was determined using daylight saving time (DST) transitions to produce a dark/light interval risk ratio; prevalence was determined using the corresponding annual crash rate in darkness for each scenario. For curve lighting, pedestrian crashes on curved roadways were examined; for motorway lighting, crashes associated with high speed roadways were examined; and for cornering light, crashes involving turning vehicles at intersections were examined. In the curve analysis, lower dark/light crash ratios were observed for curved sections of roadway compared to straight roads. In the motorway analysis, posted speed limit was the dominant predictor of this ratio for the fatal crash dataset; road function class was the dominant predictor of the ratio for the fatal/nonfatal dataset. Finally, in the intersection crash analysis, the dark/light ratio for turning vehicles was lower than for nonturning vehicles; and the ratio at intersections was lower than at non-intersections. Relative safety need was determined by combining the dark/light ratio with prevalence data to produce an idealized measure of lifesaving potential. While all three scenarios suggested a potential for safety improvement, scenarios related to high speed roadway environments showed the greatest potential. PMID- 17126279 TI - Malaria epidemiology in low-endemicity areas of the Atlantic Forest in the Vale do Ribeira, Sao Paulo, Brazil. AB - We describe a seroepidemiological survey of malaria prevalence in two areas of low endemicity: Intervales State Park and Alto Ribeira State Tourist Park (PETAR). Both are located in the Vale do Ribeira in the state of Sao Paulo, Brazil. In this study, 318 subjects from both areas had their blood analyzed for the presence of malaria parasites by thin and thick blood smears. One hundred and sixty-three (51.2%) of the subjects were from Intervales State Park and 155 (48.7%) were from PETAR. We analyzed all the samples by indirect immunofluorescent assay (IFA) to detect antibodies against asexual forms of Plasmodium vivax and Plasmodium malariae and enzyme immunosorbent assay (ELISA) to detect the presence of antibodies against circumsporozoite proteins (CSP) from P. vivax VK210, human P. vivax-like/Plasmodium simiovale, P. vivax VK247 and Plasmodium brasilianum/P. malariae. The presence of Plasmodium species was detected by polymerase chain reaction (PCR). Eighteen of the subjects analyzed had positive IFA results for IgM against P. malariae antigens, and three others were positive for P. vivax antigens. Positivity of IgG antibodies against P. vivax detected by IFA was high in samples from both Intervales State Park and PETAR (32.0% and 49.0%, respectively), while positivity for P. malariae was lower (16.0% and 19.3% in Intervales State Park and PETAR, respectively). ELISA tests showed a higher prevalence of antibodies against P. vivax VK210 (35.0%) in samples from Intervales State Park and against human P. vivax-like (29.7%) in samples from PETAR. PCR reactions revealed the presence of parasites in several of the samples analyzed. In Intervales State Park, one subject was infected by P. malariae and two by Plasmodium falciparum, while in PETAR, one subject was positive for P. falciparum and three for both P. falciparum and P. vivax parasites. The areas where these parks are located belong to the Atlantic Forest habitat, and inhabitants frequently, see monkeys. Our data suggest that monkeys may constitute a natural reservoir for malaria in both areas. PMID- 17126280 TI - Activity of azithromycin or erythromycin in combination with antimalarial drugs against multidrug-resistant Plasmodium falciparum in vitro. AB - Azithromycin, an azalide analog of erythromycin was assayed for its in vitro activity against multidrug-resistant Plasmodium falciparum K1 strain by measuring the 3H-hypoxanthine incorporation. Azithromycin caused inhibitory effects on the parasite growth with IC50 and IC90 values of 8.4+/-1.2 microM and 26.0+/-0.9 microM, respectively. Erythromycin inhibited growth of P. falciparum with IC50 and IC90 values of 58.2+/-7.7 microM and 104.0+/-10.8 microM, respectively. The activity of antimalarial drugs in combination with azithromycin or erythromycin against P. falciparum K1 were compared. Combinations of chloroquine with azithromycin or erythromycin showed synergistic effects against parasite growth in vitro. Combinations of quinine-azithromycin and quinine-erythromycin showed potentiation. Additive effects were observed in mefloquine-azithromycin and mefloquine-erythromycin combinations. Similar results were also produced by pyronaridine in combination with azithromycin or erythromycin. However, artesunate-azithromycin and artesunate-erythromycin combinations had antagonistic effects. The in vitro data suggest that azithromycin and erythromycin will have clinical utility in combination with chloroquine and quinine. The worldwide spread of chloroquine-resistant P. falciparum might inhibit the ability to treat malaria patients with chloroquine-azithromycin and chloroquine-erythromycin in areas of drug-resistant. The best drug combinations against multidrug-resistant P. falciparum are quinine-azithromycin and quinine-erythromycin. PMID- 17126282 TI - Characterization of an alternative splice variant of human nucleoside triphosphate diphosphohydrolase 3 (NTPDase3): a possible modulator of nucleotidase activity and purinergic signaling. AB - Nucleoside triphosphate diphosphohydrolase 3 (NTPDase3) is a cell surface, membrane-bound enzyme that hydrolyzes extracellular nucleotides, thereby modulating purinergic signaling. An alternatively spliced variant of NTPDase3 was obtained and analyzed. This alternatively spliced variant, termed "NTPDase3beta", is produced through the use of an alternative terminal exon (exon 11) in place of the terminal exon (exon 12) in the full-length NTPDase3, now termed "NTPDase3alpha". This results in an expressed protein lacking the C-terminal cytoplasmic sequence, the C-terminal transmembrane helix, and apyrase conserved region 5. The cDNA encoding this truncated splice variant was detected in a human lung library by PCR. Like the full-length NTPDase3alpha, the alternatively spliced NTPDase3beta was expressed in COS cells after transfection, but only the full-length NTPDase3alpha is enzymatically active and properly trafficked to the plasma membrane. However, when the truncated NTPDase3beta was co-transfected with full-length NTPDase3alpha, there was a significant reduction in the amount of NTPDase3alpha that was properly processed and trafficked to the plasma membrane as active enzyme, indicating that the truncated form interferes with normal biosynthetic processing of the full-length enzyme. This suggests a role for the NTPDase3beta variant in the regulation of NTPDase3 nucleotidase activity, and therefore the control of purinergic signaling, in those cells and tissues expressing both NTPDase3alpha and NTPDase3beta. PMID- 17126281 TI - Novel ZIP kinase isoform lacks leucine zipper. AB - Zipper-interacting protein kinase (ZIP kinase) has been thought to be involved in apoptosis and the C-terminal leucine zipper motif is important for its function. Recent studies have revealed that ZIP kinase also plays a role in regulating myosin phosphorylation. Here, we found novel ZIP kinase isoform in which the C terminal non-kinase domain containing a leucine zipper is eliminated (hZIPK-S). hZIPK-S binds to myosin phosphatase targeting subunit 1(MYPT1) similar to the long isoform (hZIPK-L). In addition, we found that hZIPK-S as well as hZIPK-L bind to myosin. These results indicate that a leucine zipper is not critical for the binding of ZIP kinase to MYPT1 and myosin. Consistently, hZIPK-S localized with stress-fibers where they co-localized with myosin. The residues 278-311, the C-terminal side of the kinase domain common to the both isoforms, is involved in the binding to MYPT1, while the myosin binding domain is within the kinase domain. These results suggest that the newly found hZIPK-S as well as the long isoform play an important role in the regulation of myosin phosphorylation. PMID- 17126283 TI - Crystal structure and ligand binding properties of the truncated hemoglobin from Geobacillus stearothermophilus. AB - A novel truncated hemoglobin has been identified in the thermophilic bacterium Geobacillus stearothermophilus (Gs-trHb). The protein has been expressed in Escherichia coli, the 3D crystal structure (at 1.5 Angstroms resolution) and the ligand binding properties have been determined. The distal heme pocket displays an array of hydrogen bonding donors to the iron-bound ligands, including Tyr-B10 on one side of the heme pocket and Trp-G8 indole nitrogen on the opposite side. At variance with the highly similar Bacillus subtilis hemoglobin, Gs-trHb is dimeric both in the crystal and in solution and displays several unique structural properties. In the crystal cell, the iron-bound ligand is not homogeneously distributed within each distal site such that oxygen and an acetate anion can be resolved with relative occupancies of 50% each. Accordingly, equilibrium titrations of the oxygenated derivative in solution with acetate anion yield a partially saturated ferric acetate adduct. Moreover, the asymmetric unit contains two subunits and sedimentation velocity ultracentrifugation data confirm that the protein is dimeric. PMID- 17126284 TI - The influence of pain on masseter spindle afferent discharge. AB - Muscle spindles provide proprioceptive feedback supporting normal patterns of motor activity and kinesthetic sensibility. During mastication, jaw muscle spindles play an important role in monitoring and regulating the chewing cycle and the bite forces generated during mastication. Both acute and chronic orofacial pain disorders are associated with changes in proprioceptive feedback and motor function. Experimental jaw muscle pain also alters the normal response of masseter spindle afferents to ramp and hold jaw movements. It has been proposed that altered motor function and proprioceptive input results from group III muscle afferent modulation of the fusimotor system which alters spindle afferent sensitivity in limb muscles. The response to nociceptive stimuli may enhance or reduce the response of spindle afferents to proprioceptive stimuli. Several experimental observations suggesting the possibility that a similar mechanism also functions in jaw muscles are presented in this report. First, evidence is provided to show that nociceptive stimulation of the masseter muscle primarily influences the amplitude sensitivity of spindle afferents with relatively little effect on the dynamic sensitivity. Second, reversible inactivation of the caudal trigeminal nuclei attenuates the nociceptive modulation of spindle afferents. Finally, functionally identified gamma motoneurons in the trigeminal motor nucleus are modulated by intramuscular injection with algesic substances. Taken together, these results suggest that pain-induced modulation of spindle afferent responses are mediated by small diameter muscle afferents and that this modulation is dependent, in part, on the relay of muscle nociceptive information from trigeminal subnucleus caudalis onto trigeminal gamma-motoneurons. The implication of these results will be considered in light of current theories on the relationship between jaw muscle pain and oral motor function. PMID- 17126285 TI - Relationship of chewing-stimulated whole saliva flow rate and salivary gland size. AB - OBJECTIVE: We have recently reported that unstimulated whole saliva flow rates (UWSFR) correlate positively with salivary gland sizes and body profiles of weight and body mass indices. In the present study, the correlations of chewing stimulated whole saliva flow rates (CWSFR) with salivary gland sizes and the body profiles were investigated, and the results were compared with those of UWSFR. DESIGN: Saliva samples were collected from 24 healthy young males and 26 females by the spitting method while chewing paraffin and the CWSFRs were measured. UWSFR and the estimated sizes of the three major salivary glands in our previous study were used. RESULTS: The CWSFRs in all subjects and in males correlated positively with UWSFR, but not in females. The CWSFRs in all subjects correlated positively with parotid and/or submandibular gland sizes, weights and body mass indices, just as with UWSFR; however, the correlation coefficients with salivary gland sizes were smaller than those of UWSFR. In contrast to the results of UWSFR, the correlation coefficients of the CWSFRs with parotid gland sizes in all subjects were larger than those with the sizes of the submandibular glands. The CWSFRs in males correlated only with parotid gland sizes, and those in females did not correlate with any of the parameters. CONCLUSIONS: The results suggest that the larger the size of the salivary glands, the greater the CWSFR, at least in males. PMID- 17126286 TI - Postnatal changes of local neuronal circuits involved in activation of jaw closing muscles. AB - Feeding behaviour in mammals changes from suckling to mastication during postnatal development and the neuronal circuits controlling feeding behaviour should change in parallel to the development of orofacial structures. In this review we discuss the location of excitatory premotor neurons for jaw-closing motoneurons (JCMNs) and postnatal changes of excitatory synaptic transmission from the supratrigeminal region (SupV) to JCMNs. We show that neurons located in SupV and the reticular formation dorsal to the facial nucleus most likely excite JCMNs. Excitatory inputs from SupV to JCMNs are mediated by activation of glutamate and glycine receptors in neonatal rats, whereas glycinergic inputs from SupV to JCMNs become inhibitory with age. We also show that the incidence of post spike afterdepolarization increases during postnatal development, whereas the amplitude and half-duration of the medium-duration afterhyperpolarization decrease with age. Such postnatal changes in synaptic transmission from SupV to JCMNs and membrane properties of JCMNs might be involved in the transition from suckling to mastication. PMID- 17126287 TI - Unilateral posterior crossbite and mastication. AB - OBJECTIVE: This study was designed to characterize masticatory-cycle morphology, and distance of the contact glide in the closing masticatory stroke, in adult subjects with uncorrected unilateral posterior crossbite (UPXB), comparing the results obtained with those obtained in a parallel group of normal subjects. STUDY DESIGN: Mandibular movements (masticatory movements and laterality movements with dental contact) were registered using a gnathograph (MK-6I Diagnostic System) during unilateral chewing of a piece of gum. Traces were recorded on the crossbite and non-crossbite sides in the crossbite group, and likewise on both sides in the non-crossbite group. RESULTS: Mean contact glide distance on the crossbite side in the UPXB group was significantly lower than in the control group (p<0.001), and mean contact glide distance on the non-crossbite side in the UPXB group was significantly lower than in the control group (p=0.042). Cycle morphology was abnormal during chewing on the crossbite side, with the frequency distribution of cycle types differing significantly from that for the noncrossbite side and that for the control group (p<0.001). CONCLUSIONS: Patients with crossbite showed alterations in both contact glide distances and masticatory cycle morphology. These alterations are probably adaptive responses allowing maintenance of adequate masticatory function despite the crossbite. PMID- 17126288 TI - The influence of masseter activity on rat mandibular growth. AB - Many studies have shown that mandibular and condylar growth is affected by compressive forces on mandibular bone and the condyle. It has been reported that chondroblastic differentiation and proliferation in chondrocytes play important roles in condylar growth. However, the influence of reduced compressive force on chondroblastic proliferation and mandibular bone formation is not fully understood. Thirty-six 3-week-old male Wistar rats were used in this study. In the experimental group, the masseter muscles were bilaterally resected to evaluate the influence of masticatory force on mandibular and condylar bone morphology. Six weeks after the operation, while the rats were in the pubertal growth stage, lateral X-rays were taken to analyze the skeletal pattern of the mandible. The form of the condyle and the thickness of the chondroblastic layers were evaluated by toluidine blue staining. Chondroblastic proliferation was identified by insulin-like growth factor-1 receptor (IGF-1r) immunostaining and bone resorption of the condyle was assessed by measuring tartrate-resistant acid phosphatase (TRAP) activity. Lateral X-rays of the mandible showed that rats in the experimental group tended to have large mandibular plane angles. The chondroblastic layer in the condyles of the experimental group rats was thinner than in the control group. The expression of IGF-1r immunopositive cells in the experimental group was significantly lower than in the control chondrocytes, and the number of TRAP-positive cells was significantly higher in the condylar bone of the experimental group. We conclude that masseter muscle activity is closely related to mandibular morphology during growth. PMID- 17126289 TI - Approach and avoidance in fear of spiders. AB - We examined attitudes towards spiders by employing an Approach-Avoidance Task, in which participants respond to pictures by pulling a joystick towards themselves or by pushing it away from themselves. For spider fearfuls, this stimulus response assignment is either compatible (push spiders away) or incompatible (pull spiders closer). Specific compatibility effects were found: compared to non anxious controls and control pictures, highly spider fearful participants responded to spider pictures more quickly by pushing than by pulling, even when picture contents was task-irrelevant. Moreover, compatibility effects predicted fear-related behavior independently of questionnaires. Potential applications, extensions, and limitations of the findings are discussed. PMID- 17126290 TI - Blockade of TRAIL pathway ameliorates HBV-induced hepatocyte apoptosis in an acute hepatitis model. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) may play important roles during hepatitis B virus (HBV) infection. In this study, we used a recombinant human soluble death receptor 5 (sDR5) to explore its effect in a mouse model of HBV-induced acute hepatitis. By measuring blood transaminase activity and hepatocyte apoptosis, we found that sDR5 could alleviate liver damage by blocking TRAIL-induced apoptosis of HBV-transfected hepatocytes. sDR5 injection at 16 mg/kg 24h before HBV transfection was the most effective. Additionally, we showed that sDR5 was equally effective in protecting liver injury as the Stronger Neo-Minophagen C (SNMC), a commonly used drug for patients with liver diseases. Thus, sDR5 represents a potential novel therapeutic drug for patients with fulminant hepatitis. PMID- 17126291 TI - A transgenic rat with the human ATTR V30M: a novel tool for analyses of ATTR metabolisms. AB - Amyloidogenic transthyretin (ATTR) is the pathogenic protein of familial amyloidotic polyneuropathy (FAP). To establish a tool for analyses of ATTR metabolisms including after liver transplantations, we developed a transgenic rat model expressing human ATTR V30M and confirmed expressions of human ATTR V30M in various tissues. Mass spectrometry for purified TTR revealed that rat intrinsic TTR and human ATTR V30M formed tetramers. Congo red staining and immunohistochemistry revealed that nonfibrillar deposits of human ATTR V30M, but not amyloid deposits, were detected in the gastrointestinal tracts of the transgenic rats. At 24h after liver transplantation, serum human ATTR V30M levels in transgenic rats that received livers from normal rats became lower than detectable levels. These results thus suggest that this transgenic rat may be a useful animal model which analyzes the metabolism of human ATTR V30M including liver transplantation studies. PMID- 17126292 TI - Beta-galactosidase (Escherichia coli) has a second catalytically important Mg2+ site. AB - It is shown here that Escherichia coli beta-galactosidase has a second Mg2+ binding site that is important for activity. Binding of Mg2+ to the second site caused the k(cat) (with oNPG as the substrate) to increase about 100 s(-1); the Km was not affected. The Kd for binding the second Mg2+ is about 10(-4)M. Since the concentration of free Mg2+ in E. coli is about 1-2 mM, the second site is physiologically significant. Non-polar substitutions (Ala or Leu) for Glu-797, a residue in an active site loop, eliminated the k(cat) increase. This indicates that the second Mg2+ site is near to Glu-797. The Ki values of transition state analogs were decreased by small but statistically significant amounts when the second Mg2+ site was occupied and Arrhenius plots showed that less entropic activation energy is required when the second site is occupied. These inhibitor and temperature results suggest that binding of the second Mg2+ helps to order the active site for stabilization of the transition state. PMID- 17126293 TI - The neuropeptide SIFamide modulates sexual behavior in Drosophila. AB - The expression of Drosophila neuropeptide AYRKPPFNGSIFamide (SIFamide) was shown by both immunohistology and in situ hybridization to be restricted to only four neurons of the pars intercerebralis. The role of SIFamide in adult courtship behavior in both sexes was studied using two different approaches to perturb the function of SIFamide; targeted cell ablation and RNA interference (RNAi). Elimination of SIFamide by either of these methods results in promiscuous flies; males perform vigorous and indiscriminant courtship directed at either sex, while females appear sexually hyper-receptive. These results demonstrate that SIFamide is responsible for these behavioral effects and that the four SIFamidergic neurons and arborizations play an important function in the neuronal circuitry controlling Drosophila sexual behavior. PMID- 17126294 TI - Neuron-specific TGF-beta signaling deficiency results in retinal detachment and cataracts in mice. AB - We generated a mouse model (cKO) with a conditional deletion of TGF-beta signaling in the retinal neurons by crossing TGF-beta receptor I (TGF-beta RI) floxed mice with nestin-Cre mice. Almost all of the newborn cKO mice had retinal detachment at the retinal pigment epithelium (RPE)/photoreceptor layer junction of the neurosensory retina (NSR). The immunostaining for chondroitin-6-sulfate showed a very weak reaction in cKO mice in contrast to intense staining in the photoreceptor layer in wild-type mice. Macroscopic cataracts, in one or both eyes, were observed in 50% of the mice by 6 months of age, starting as early as the first month after birth. The cKO mouse model demonstrates that the TGF-beta signaling deficiency in retinal cells leads to decreased levels of chondroitin sulfate proteoglycan in the retinal interphotoreceptor matrix. This in turn causes retinal detachment due to the loss of adhesion of the NSR to RPE. PMID- 17126295 TI - Selenazoles (selenium compounds) facilitate survival of cultured rat pheochromocytoma PC12 cells after serum-deprivation and stimulate their neuronal differentiation via activation of Akt and mitogen-activated protein kinase, respectively. AB - The activation of extracellular receptor kinase (ERK) is one of the checkpoints to assess the activation of the classical Ras/mitogen-activated protein kinase (MAPK) cascade. Therefore, we tested more than 100 selenium-containing compounds for their ability to activate the MAPK signal pathway. Among them, we found that three selenazoles, 5-chloroacetyl-2-piperidino-1,3-selenazole (CS1), 5 chloroacetyl-2-morpholino-1,3-selenazole (CS2), and 5-chloroacetyl-2 dimethylamino-1,3-selenazole (CS3), induced the phosphorylation of ERK. These compounds also enhanced the phosphorylation of Akt, a signal transducing protein kinase for cell survival; and this phosphorylation was followed by suppression of cell death, thus suggesting that they had anti-apoptotic effects. Moreover, CSs 1 3 induced neurite outgrowth and facilitated the expression of neurofilament-M of PC12 cells, demonstrating that they induced neuronal differentiation of these cells. On the other hand, the CS-induced phosphorylation of MAPK was enhanced by buthionine sulfoximine (BSO), an activator of protein tyrosine phosphatases (PTPs), but inhibited by N-acetyl-l-cysteine (NAC), an inhibitor of receptor tyrosine kinase. These results imply that activation of some receptor tyrosine kinase(s) is involved in the mechanism of action of CSs 1-3. The activation of MAPK by CSs 1-3 was suppressed by U0126, a MEK inhibitor, but not by K252a, an inhibitor of TrkA; AG1478, an antagonist of epidermal growth factor receptor (EGFR); or by pertussis toxin. These results demonstrate that the CS-induced phosphorylation of Akt and MAP kinase (receptor tyrosine kinase(s)-MEK1/2-ERK1/2) cascades was responsible for suppression of apoptosis and facilitation of neuronal differentiation of PC12 cells, respectively. Our results suggest that CSs 1-3 are promising candidates as neuroprotective and/or neurotrophic agents for the treatment of various neurodegenerative neurological disorders. PMID- 17126296 TI - Evidence for an immune function of lepidopteran silk proteins. AB - Hemolymph coagulation stops bleeding and protects against infection. Clotting factors include both proteins that are conserved during evolution as well as more divergent proteins in different species. Here we show that several silk proteins also appear in the clot of the greater wax moth Galleria mellonella. RT-PCR analysis reveals that silk proteins are expressed in immune tissues and induced upon wounding in both Galleria and Ephestia kuehniella, a second pyralid moth. Our results support the idea that silk proteins were co-opted for immunity and coagulation during evolution. PMID- 17126297 TI - Is there any effect of renal failure on the hepatic regeneration capacity following partial hepatectomy in rats? AB - The effects of renal dysfunction on liver regeneration capacity have not been fully elucidated before, although many patients with renal failure are subjected to hepatectomy due to hepatobiliary diseases. In this study, we sought to determine the effects of renal dysfunction on the hepatic regeneration capacity using rat chronic renal failure model. After establishing chronic renal failure (CRF group) by semi-total renal resection, the rats were subjected to 70% partial hepatectomy (PHx). Rats without renal failure were used as control (Sham group). The hepatic regeneration rate, histology of the liver, clearance of indocyanine green into the bile, and the expression of hepatic regeneration-associated genes in the liver were evaluated. The hepatic regeneration rate was lower in CRF group as compared to Sham group on day 1 after PHx. Mitotic index evaluated by histologic examination on day 1 after PHx was also significantly lower in CRF group. However, no difference in these indices was observed on day 2 and 7 between Sham and CRF. Indocyanine green clearance rate was almost identical between Sham and CRF on day 7 following PHx. The baseline expressions of the hepatic regeneration-associated genes, such as IL-6, TNF-alpha, HGF, c-fos, and c jun, in the liver of CRF were significantly lower than those of Sham. However, the rate of upregulation of these genes was not significantly different between Sham and CRF. These results clearly demonstrate that the renal dysfunction, although initially delays the onset, does not suppress the total hepatic regeneration capacity following partial hepatectomy. The function of the regenerated liver on day 7 after PHx also was not different. Our results provide a possibility that the hepatectomy can be indicated even for the patient with a chronic renal failure. PMID- 17126298 TI - Tyrosine 311 is phosphorylated by c-Abl and promotes the apoptotic effect of PKCdelta in glioma cells. AB - In this study we characterized the phosphorylation of tyrosine 311 and its role in the apoptotic function of PKCdelta in glioma cells. We found that c-Abl phosphorylated PKCdelta on tyrosine 311 in response to H2O2 and that this phosphorylation contributed to the apoptotic effect of H2O2. In contrast, Src, Lyn, and Yes were not involved in the phosphorylation of tyrosine 311 by H2O2. A phosphomimetic PKCdelta mutant, in which tyrosine 311 was mutated to glutamic acid (PKCdeltaY311E), induced a large degree of cell apoptosis. Overexpression of the PKCdeltaY311E mutant induced the phosphorylation of p38 and inhibition of p38 abolished the apoptotic effect of the PKCdelta mutant. These results suggest an important role of tyrosine 311 in the apoptotic function of PKCdelta and implicate c-Abl as the kinase that phosphorylates this tyrosine. PMID- 17126299 TI - Protein tyrosine phosphorylation of a heparin-binding sperm membrane mitogen (HBSM) is associated with capacitation and acrosome reaction. AB - Protein tyrosine phosphorylation in spermatozoa is associated with epididymal maturation and though to be central for attainment of a capacitated state and expression of hyperactivated motility. Heparin, the most highly sulfated glycosaminoglycans, was also the most potent at stimulating the acrosomal reaction in bovine epididymal spermatozoa. Studies using radiolabeled inorganic phosphate showed 11-fold increase (32)Pi incorporation in heparin-binding sperm membrane protein (HBSM) during spermatozoal capacitation, and the phosphorylation occurs at the tyrosine residue. Epididymal spermatozoa were induced to undergo capacitation and acrosome reaction by 70% when the cells were incubated in BWW medium supplemented with heparin. The spermatozoa pre-treated with anti-HBSM antibody showed 46% reduction in the hyperactivated motility and lowers the acrosome reaction. This was confirms by measuring the hydrolysis of benzoyl-l arginine ethyl ether (BAEE) by the acrosomal enzyme; acrosin. The preliminary finding suggests that HBSM may play an important role in the sperm capacitation and acrosome reaction. PMID- 17126300 TI - Actin cytoskeletons regulate the stretch-induced increase of Ca2+ current in human gastric myocytes. AB - Using the whole-cell and single channel recording techniques, the influence of actin cytoskeletons on L-type Ca2+ current was investigated in human gastric smooth muscle cells. In isotonic condition, an actin depolymerizer cytochalasin D (Cyt-D) markedly decreased the whole-cell current (I(Ba)) without changing steady state voltage dependency and single channel conductance. Intracellular dialysis of phalloidin, an actin polymerizer, significantly increased the I(Ba). Hypotonic stretch (222 mOsm/L) of the myocytes increased the I(Ba), and Cyt-D significantly inhibited the I(Ba) increase by the stretch. Phalloidin was without effect on the I(Ba) increase by the stretch. Phalloidin antagonized the Cyt-D inhibition of the stretch-induced I(Ba) increase. Neither heterotrimeric G protein modifiers (GTPgammaS and GDPbetaS) nor rho GTPase inhibitor (C3 exoenzyme) influenced the stretch-induced responses. These results reveal that the integrity of the actin cytoskeleton is an important factor which determines the activity of L-type Ca2+ channels and a response to stretch. PMID- 17126301 TI - Activated beta-catenin in the novel human parathyroid tumor cell line sHPT-1. AB - Misregulation of the Wnt/beta-catenin signalling pathway is involved in the development and progression of many cancers. Recently, we presented evidence for aberrant accumulation of non-phosphorylated (stabilized) beta-catenin in benign parathyroid tumors from patients with primary hyperparathyroidism (pHPT) or HPT secondary to uremia (sHPT). Here we have used a human parathyroid hormone (PTH) producing cell line (sHPT-1), established from a hyperplastic parathyroid gland removed at operation of a patient with sHPT, to further investigate the potential importance of beta-catenin in parathyroid tumorigenesis. Our studies demonstrate that efficient and specific knockdown of beta-catenin by small interfering RNA (siRNA) markedly decreased endogenous beta-catenin transcriptional activity as well as expression of the Wnt/beta-catenin target genes cyclin D1 and c-myc, known to be overexpressed in a substantial fraction of parathyroid tumors. Furthermore, siRNA to beta-catenin inhibited cellular growth and induced cell death. Growth and survival of the parathyroid tumor cells are thus dependent on maintained expression level of beta-catenin. The Wnt/beta-catenin signalling pathway, and beta-catenin in particular, presents a potential therapeutic target for HPT. PMID- 17126302 TI - Activation of PPARalpha lowers synthesis and concentration of cholesterol by reduction of nuclear SREBP-2. AB - To elucidate the mechanisms underlying the cholesterol lowering effects of PPARalpha agonists we investigated key regulators of cholesterol synthesis and uptake in rats and in the rat hepatoma cell line Fao after treatment with the PPARalpha agonists clofibrate and WY 14,643, respectively. In rat liver as well as in Fao cells, PPARalpha activation led to a decrease of transcriptionally active nuclear SREBP-2. mRNA concentrations of the key regulators of SREBP processing, Insig-1 in rat liver and Insig-1 and Insig-2a in Fao cells, were increased upon PPARalpha activation. Thus we suggest, that the observed reduction of the amount of nuclear SREBP-2 was due to an inhibition of the processing of the precursor protein. Both, in rat liver and in Fao cells, mRNA concentrations of the SREBP-2 target genes HMG-CoA reductase (EC1.1.1.34) and LDL receptor were reduced after treatment with the PPARalpha agonists. Furthermore, treatment of Fao cells with WY 14,643 reduced cholesterol synthesis. As a result, the amount of total cholesterol in liver, plasma and lipoproteins of clofibrate treated rats and in WY 14,643 treated Fao cells was decreased compared to control animals and cells, respectively. In conclusion, we could show a novel link between PPARalpha and cholesterol metabolism by demonstrating that PPARalpha activation lowers cholesterol concentration by reducing the abundance of nuclear SREBP-2. PMID- 17126303 TI - Deep brain stimulation in movement and psychiatric disorders. AB - Deep brain stimulation (DBS) is the most focal and invasive of the electromagnetic brain stimulation therapies. A subcutaneous pulse generator provides continuous stimulation of circumscribed brain tissue via a multicontact microelectrode that terminates within its target. The result is an adjustable, reversible, and specific therapy. Despite limited understanding of its mechanisms of action, DBS efficacy has been established in several movement disorders, and promising reports have emerged for Tourette syndrome, obsessive-compulsive disorder, and major depression. Deep brain stimulation may prove to be a reasonable option for severely ill and treatment-resistant patients who otherwise have limited therapeutic options and a poor prognosis. PMID- 17126304 TI - Relationship of movements and behaviors to Group A Streptococcus infections in elementary school children. AB - BACKGROUND: Pediatric Autoimmune Neuropsychiatric Disorders Associated with Streptococcus (PANDAS) research is based on the hypothesis that infections trigger changes in behavior and movement in children. METHODS: We enrolled 693 children (ages 3 to 12 years) into a systematic, longitudinal study. Data were collected monthly for 8 months (October-May) to determine point prevalence of Group A Streptococcal (GAS) infections, tics, behavior, and choreiform movements. Simultaneous throat cultures were obtained, and relational analyses were made between GAS and movement/observation ratings. RESULTS: Combined behavior/GAS associations (concurrent with or 3 subsequent months to GAS) revealed a strong relationship, relative risk (RR) of 1.71 (p < .0001). Detailed analysis revealed that balance/swaying and non-tic grimacing were responsible for a significant proportion of this association (RR = 2.92, p < .0001). A strong seasonal pattern was found, with fall being more significant for GAS infections and observation ratings (p < .0001) compared with winter/spring. Children with repeated streptococcus (n = 64) showed higher rates of behavior and distal choreiform observations (p = .005). CONCLUSIONS: Motor/behavior changes were noted to occur in relationship to positive GAS culture with support that repeated GAS increases risk. PMID- 17126305 TI - Chemokines in the MPTP model of Parkinson's disease: absence of CCL2 and its receptor CCR2 does not protect against striatal neurodegeneration. AB - Recent studies have invoked inflammation as a major contributor to the pathogenesis of Parkinson's disease (PD). We determined the role of members of the chemokine system, key inflammatory mediators, in PD pathogenesis. In the MPTP model of murine PD, several chemokines, including CC chemokine ligand 2 (CCL2, Monocyte Chemoattractant Protein-1) and CCL3 (Macrophage Inflammatory Protein 1alpha), were upregulated in the striatum and the ventral midbrain. Astrocytes were the predominant source of CCL2 and CCL3 in the striatum and the substantia nigra, and dopaminergic neurons in the substantia nigra constitutively expressed these two chemokines. MPTP treatment resulted in decreased CCL2 expression and increased CCL3 expression in the surviving dopaminergic neurons. Because we found that CCL2 induced production of TNF-alpha in microglial cells, a cytokine known to play a detrimental role in PD, we anticipated that deletion of the genes encoding CCL2 and CCR2, its major receptor, would confer a protective phenotype. However, MPTP-induced striatal dopamine depletion was comparable in double knockout and wild-type mice. Our results demonstrate that chemokines such as CCL2 are induced following MPTP treatment, but that at least within the context of this PD model, the absence of CCL2 and CCR2 does not protect against striatal dopamine loss. PMID- 17126306 TI - Analysis of transcriptional modulation of the presenilin 1 gene promoter by ZNF237, a candidate binding partner of the Ets transcription factor ERM. AB - DNA sequences required for the expression of the human presenilin 1 (PS1) gene have been identified between -118 and +178 flanking the major initiation site (+1) mapped in SK-N-SH cells. Several Ets sites are located both upstream as well as downstream from the +1 site, including an Ets motif present at -10 that controls 90% of transcription in SK-N-SH cells. However, in SH-SY5Y cells, transcription initiates further downstream and requires an alternative set of promoter elements including a +90 Ets motif. Ets2, ER81, ERM and Elk1 were identified by yeast one-hybrid selection in a human brain cDNA library using the 10 Ets motif as a bait. We have shown that ERM recognizes specifically Ets motifs on the PS1 promoter located at -10 as well as downstream at +90, +129 and +165 and activates PS1 transcription with promoter fragments whether or not they contain the -10 Ets site. We have now searched for ERM interacting proteins by yeast two-hybrid selection in a human brain cDNA library using the C-terminal 415 amino acid of ERM as a bait. One of the interacting proteins was ZNF237, a member of the MYM gene family. It is widely expressed in different tissues in eukaryotes under several forms derived by alternative splicing, including a large 382 amino acid form containing a single MYM domain, and 2 shorter forms of 208 and 213 amino acids respectively that do not. We show that both the 382 as well as the 208 amino acid forms are expressed in SK-N-SH cells but not in SH-SY5Y cells. Both forms interact with ERM and repress the transcription of PS1 in SH-SY5Y cells. The effect of both C-terminal and N-terminal deletions indicates that the N-terminal 120 amino acid region is required for interaction with ERM in yeast, and furthermore single amino acid mutations show that residues 112 and 114 play an important role. The repression of transcription in SH-SY5Y cells also appears to require the N-terminal potion of ZNF237 and was affected by mutation of the amino acid 112. Data from electrophoretic mobility shift assays indicate that ERM and possibly ZNF237 interact with a fragment of the PS1 promoter. PMID- 17126307 TI - Reperfusion kinase phosphorylation is essential but not sufficient in the mediation of pharmacological preconditioning: Characterisation in the bi-phasic profile of early and late protection. AB - OBJECTIVE: Pharmacological preconditioning (PPC) triggers early (ePPC) and delayed protection (dPPC), occurring within 1 h or after 24 h following the preconditioning stimulus, respectively, through recruitment of protein kinase signalling. Angiotensin II (ATII) is a recognised trigger of PPC, recruiting kinases and transcription factors known to be involved in both phases of protection. Our objectives were to determine whether ATII is capable of triggering dPPC and whether recruitment of pro-survival kinases, Akt and extracellular signal-regulated kinase (ERK), following the injurious ischaemic insult is essential for the mediation of PPC. METHODS: In a mouse Langendorff model of ischaemia/reperfusion injury, we undertook to determine whether ATII triggers both ePPC and dPPC. Western blot analysis was used to determine kinase phosphorylation at reperfusion, and kinase inhibitors wortmannin and PD98059 were used to ascertain the significance of kinase regulation. RESULTS: We demonstrated that ATII triggered PPC with attenuation of infarction at 1 and 24 h (19+/-4% and 25+/-4% versus control, 35+/-4% of risk zone, p < 0.05), consistent with the ePPC and dPPC time-course. This bi-phasic protection was associated with significant post-ischaemic phosphorylation of both Akt and ERK within the first 5 min of reperfusion. Akt and ERK phosphorylation was increased following ePPC by 4.5+/ 0.5 and 1.9+/-0.6 fold, respectively (p < 0.001), and dPPC by 24+/-2.0 and 2.1+/ 0.1 fold, respectively (p < 0.001). Both wortmannin and PD98059 administered during reperfusion ameliorated the phosphorylation of Akt and ERK and abrogated the resistance to infarction resulting from both ePPC and dPPC (33+/-3% and 35+/ 4%, respectively, versus controls 33+/-4% and 33+/-5%, p = NS). There was no evidence of augmented phosphorylation of either p38 kinase or JNK at either time point. CONCLUSION: We demonstrate that PPC results in a clearly delineated time course of bi-phasic protection against injurious ischemic injury that is correlated with reperfusion kinase phosphorylation of both Akt and ERK. These data indicate a novel mechanism of early and particularly delayed preconditioning. PMID- 17126308 TI - Factor V Leiden and apolipoprotein E genotypes in severe femoropopliteal atherosclerosis with restenosis. AB - BACKGROUND: The role of factor V (Leiden) mutation, thrombophilia, and apolipoprotein E (apoE) alleles in the pathogenesis of accelerated atherosclerosis and restenosis was studied in patients requiring reoperation within five years after femoropopliteal angioplasty with artificial grafts. METHODS: One hundred ninety-eight consecutive patients with femoropopliteal atherosclerotic disease, reoperated for restenosis were contacted by phone and 100 of them returned for laboratory and clinical work-up. In addition to clinical evaluation and routine laboratory investigations, parameters of lipoprotein metabolism, factor V (Leiden) mutation and apolipoprotein E (apoE) allele were studied by PCR amplification of DNA and endonuclease digestion techniques. RESULTS: A significantly higher incidence of factor V (Leiden) mutation was found in patients with atherosclerosis and restenosis, compared to 445 healthy blood donors (13/200, 6.5% vs. 34/890, 3.8%, p=0.0379). Distribution of the alleles of the apolipoprotein E (apoE) gene was different, when the patients were compared to 372 controls; however, the difference only approached the level of statistical significance (25/200, 12.5% vs. 56/744, 7.5%, p=0.0515). Comparing the two groups, the number of epsilon4 allele carriers was significantly higher among patients with restenosis (25/100, 25% vs. 53/272, 14%, p=0.0147). CONCLUSION: Factor V (Leiden) mutation may influence the progression of atherosclerosis and the development of restenosis after revascularization in patients with accelerated femoropopliteal atherosclerosis. Further investigation is needed whether long-term anticoagulation has an impact or not on the course of disease in such cases. ApoE epsilon4 allele should be screened in patients with femoropopliteal atherosclerosis, because it indicates a faster progression of atherosclerosis and may predict restenosis after revascularization procedure. PMID- 17126309 TI - AMI is associated with polymorphisms in the NOS3 and FGB but not in PAI-1 genes in young adults. AB - BACKGROUND: We investigated the relationship between NOS3, FGB and PAI-1 polymorphisms and endothelial dysfunction and risk factors for acute myocardial infarction (AMI) in young adults. METHODS: Endothelial function was measured by response to flow mediated vasodilation (FMV) and induced by nitrate (FMN). Biochemical parameters were measured by standard enzymatic methods and plasma total nitrate was analyzed by the NOA system. NOS3 (T-786C, G894T and intron 4A/B STR), FGB (C-148T and G-455A) and PAI-1 (4G/5G) polymorphisms were determined by PCR-RFLP. RESULTS: Concentrations of total and LDL cholesterol, apo B, triglycerides, nitrate, PAI-1 and fibrinogen were higher and apo AI, HDL cholesterol and FMV were lower in AMI patients than in controls (p<0.001). PAI-1 (p<0.001) but not nitrate was higher in AMI patients with low response to FMV. NOS3 T-786C and FGB C-148T polymorphisms were associated with AMI (p<0.050). NOS3 T-786C was also related to hypertension (p=0.049). NOS3 intron 4A/B STR was associated with increased concentrations of total cholesterol and apo B. NOS3 786TT/894GT haplotype was associated with increased FMV (p=0.018) than the other haplotypes. CONCLUSIONS: Our data suggest NOS3 and FGB polymorphisms are associated with AMI. NOS3 is also related to hypertension, endothelial dysfunction and variation on serum cholesterol in young adults with AMI. PMID- 17126310 TI - Performance characteristics of seven automated thyroxine and T-uptake methods. AB - BACKGROUND: Estimates of the free thyroxine concentration can be made using measurements of thyroxine (T4) and the thyroid hormone binding ratio (THBR, usually reported as T-uptake) according to the formula free thyroxine index (FTI)=T4xTHBR. We evaluated the performance characteristics of 7 pairs of automated T4 and T-uptake methods. METHODS: We evaluated the Architect i2000, AxSYM, ADVIA Centaur, UniCel DxI 800, Immulite 2000, Modular E170 and Vitros ECi methods for T4 and T-uptake. Imprecision was assessed by duplicate determinations on 3 levels of quality control material per run, 2 runs per day, on 5 separate days. Method comparison studies were performed with 205 patient samples and 68 samples from subjects in the second or third trimester of pregnancy. Results for both imprecision and method comparison were converted to the THBR as outlined by each method's manufacturer. RESULTS: Overall CVs for T4 and THBR methods were 3.2 8.9% and 1.2-6.7%, respectively. T4 methods generally agreed well. The THBR methods did not agree nearly as well. Comparison of FTI results with free T4 by equilibrium dialysis showed good correlation but different slopes. CONCLUSIONS: All methods show adequate precision but the THBR and FTI results are not well standardized. PMID- 17126311 TI - Clinical, biochemical and molecular diagnosis of a compound homozygote for the 254 bp deletion-8 bp insertion of the APRT gene suffering from severe renal failure. AB - OBJECTIVE: To determine the type of mutation in a patient with clinical diagnosis of suspected APRT deficiency. DESIGN AND METHODS: A 51-year-old male patient, with a clinical history of two prior episodes of renal colic with urinary stone excretion (reported as uric acid stones in the first episode and as calcium oxalate stones in the second), was admitted to the hospital with severe non oliguric renal failure (1.06 mmol/L serum creatinine), severe hyponatremia (114 mmol/L Na(+)), metabolic acidosis (14 mmol/L HCO(3)(-)) and uricemia in the normal range. Abnormalities at renal scan and persistency of severe renal failure required to start haemodialysis. Results of renal biopsy prompted us to undertake a biochemical and molecular biological evaluation of the patient for suspected adenine phosphoribosyltransferase (APRT) deficiency. RESULTS: HPLC analysis of serum and urine, for determining purine derivative profile, showed the pathological presence of adenine in both biological fluids (3.57 micromol/L and 7.11 micromol/mmol creatinine in serum and urine, respectively; not detectable in both fluids in healthy controls). APRT assay in a sample of patient hemolysate showed no detectable activity of the enzyme (25.56+/-9.55 U/L red blood cells in control healthy subjects). Molecular biological analysis of the amplified APRT gene revealed that the patient harboured in exon 3 a homozygous 254 bp deletion-8 bp insertion, previously described only once in a compound heterozygote. Analysis of the patient family showed that heterozygotes for this APRT gene mutation, in spite of a 69% lower APRT enzymatic activity than that of healthy subjects, had no detectable adenine concentrations in both serum and urine. CONCLUSIONS: Results of the first patient harbouring the homozygous 254 bp deletion-8 bp insertion of the APRT gene strongly indicated that definitive diagnosis of APRT deficiency (often under or misdiagnosed) would require a combined clinical, biochemical and molecular biological evaluation. PMID- 17126312 TI - Novel homogenous time-resolved fluorometric RT-PCR assays for quantification of PSA and hK2 mRNAs in blood. AB - OBJECTIVES: The purpose of this study was to design, validate, and optimize internally standardized real-time quantitative RT-PCR assays and to identify and avoid problems with assay reliability and examine the impact of an exogenous internal standard. DESIGN AND METHODS: The model system consisted of internally standardized quantitative real-time RT-PCR assays specific for PSA and hK2 mRNA based on time-resolved fluorometric detection of lanthanide chelates. RESULTS: Reproducibility was best when large copy numbers (>5000 per milliliter blood) were analyzed. Addition of an exogenous target-mimicking internal standard had no significant effect on the reproducibility of the method, but increased the calculated copy numbers by an average of 2-fold. CONCLUSIONS: We developed an internally standardized, specific and reproducible real-time RT-PCR analysis method for PSA and hK2 mRNA in circulating cells in the bloodstream. Both PSA and hK2 assays are sufficiently sensitive to detect two LNCaP cells per milliliter whole blood. PMID- 17126313 TI - Distinct modulation of alkaline phosphatase isoenzymes by 17beta-estradiol and xanthohumol in breast cancer MCF-7 cells. AB - OBJECTIVES: To examine the effect of 17beta-estradiol and xanthohumol in alkaline phosphatase (ALP) expression and activity in breast cancer MCF-7 cells. DESIGN AND METHODS: ALP isoenzymes expression was evaluated by RT-PCR and Western blotting. ALP activity was measured by spectrophotometry. Cell proliferation and apoptosis were examined by MTT and immunostaining for KI67 and TUNEL, respectively. RESULTS: ALP isoenzymes expression and activity were decreased by 1 nM 17beta-estradiol. Pure estrogenic antagonist (ICI 182,780) reversed 17beta estradiol-inhibiting effect in TNS-ALP expression. RNA and protein expression of IALP, but not TNS-ALP, was also decreased by incubation with 10 microM xanthohumol (IC(50)) and was accompanied by a significant reduction in ALP activity. Treatment with 17beta-estradiol enhanced cell proliferation and decreased apoptosis. Conversely, xanthohumol incubation inhibited cell viability and apoptosis. CONCLUSION: Estrogens and xanthohumol differently modulate ALP isoenzymes. ALP loss associated with increased cell proliferation. Modulation of this enzyme by 17beta-estradiol and xanthohumol might provide therapeutic strategies against hormone-dependent breast cancer. PMID- 17126314 TI - A visual probe localization and calibration system for cost-effective computer aided 3D ultrasound. AB - The 3D ultrasound systems produce much better reproductions than 2D ultrasound, but their prohibitively high cost deprives many less affluent organization this benefit. This paper proposes using the conventional 2D ultrasound equipment readily available in most hospitals, along with a single conventional digital camera, to construct 3D ultrasound images. The proposed system applies computer vision to extract position information of the ultrasound probe while the scanning takes place. The probe, calibrated in order to calculate the offset of the ultrasound scan from the position of the marker attached to it, is used to scan a number of geometrical objects. Using the proposed system, the 3D volumes of the objects were successfully reconstructed. The system was tested in clinical situations where human body parts were scanned. The results presented, and confirmed by medical staff, are very encouraging for cost-effective implementation of computer-aided 3D ultrasound using a simple setup with 2D ultrasound equipment and a conventional digital camera. PMID- 17126315 TI - Meltrin beta expressed in cardiac neural crest cells is required for ventricular septum formation of the heart. AB - The heart is divided into four chambers by membranous septa and valves. Although evidence suggests that formation of the membranous septa requires migration of neural crest cells into the developing heart, the functional significance of these neural crest cells in the development of the endocardial cushion, an embryonic tissue that gives rise to the membranous appendages, is largely unknown. Mice defective in the protease region of Meltrin beta/ADAM19 show ventricular septal defects and defects in valve formation. In this study, by expressing Meltrin beta in either endothelial or neural crest cell lineages, we showed that Meltrin beta expressed in neural crest cells but not in endothelial cells was required for formation of the ventricular septum and valves. Although Meltrin beta-deficient neural crest cells migrated into the heart normally, they could not properly fuse the right and left ridges of the cushion tissues in the proximal outflow tract (OT), and this led to defects in the assembly of the OT and AV cushions forming the ventricular septum. These results genetically demonstrated a critical role of cardiac neural crest cells expressing Meltrin beta in triggering fusion of the proximal OT cushions and in formation of the ventricular septum. PMID- 17126317 TI - Rescue of cytochrome P450 oxidoreductase (Por) mouse mutants reveals functions in vasculogenesis, brain and limb patterning linked to retinoic acid homeostasis. AB - Cytochrome P450 oxidoreductase (POR) acts as an electron donor for all cytochrome P450 enzymes. Knockout mouse Por(-/-) mutants, which are early embryonic (E9.5) lethal, have been found to have overall elevated retinoic acid (RA) levels, leading to the idea that POR early developmental function is mainly linked to the activity of the CYP26 RA-metabolizing enzymes (Otto et al., Mol. Cell. Biol. 23, 6103-6116). By crossing Por mutants with a RA-reporter lacZ transgene, we show that Por(-/-) embryos exhibit both elevated and ectopic RA signaling activity e.g. in cephalic and caudal tissues. Two strategies were used to functionally demonstrate that decreasing retinoid levels can reverse Por(-/-) phenotypic defects, (i) by culturing Por(-/-) embryos in defined serum-free medium, and (ii) by generating compound mutants defective in RA synthesis due to haploinsufficiency of the retinaldehyde dehydrogenase 2 (Raldh2) gene. Both approaches clearly improved the Por(-/-) early phenotype, the latter allowing mutants to be recovered up until E13.5. Abnormal brain patterning, with posteriorization of hindbrain cell fates and defective mid- and forebrain development and vascular defects were rescued in E9.5 Por(-/-) embryos. E13.5 Por(-/-); Raldh2(+/-) embryos exhibited abdominal/caudal and limb defects that strikingly phenocopy those of Cyp26a1(-/-) and Cyp26b1(-/-) mutants, respectively. Por(-/-); Raldh2(+/-) limb buds were truncated and proximalized and the anterior-posterior patterning system was not established. Thus, POR function is indispensable for the proper regulation of RA levels and tissue distribution not only during early embryonic development but also in later morphogenesis and molecular patterning of the brain, abdominal/caudal region and limbs. PMID- 17126316 TI - Engrailed controls the organization of the ventral nerve cord through frazzled regulation. AB - In Drosophila, the ventral nerve cord (VNC) architecture is built from neuroblasts that are specified during embryonic development, mainly by transcription factors. Here we show that Engrailed, a homeodomain transcription factor known to be involved in the establishment of neuroblast identity, is also directly implicated in the regulation of axonal guidance cues. Posterior commissures (PC) are missing in engrailed mutant embryos, and axonal pathfinding defects are observed when Engrailed is ectopically expressed at early stages, prior to neuronal specification. We also show that frazzled, enabled, and trio, all of which are potential direct targets of Engrailed and are involved in axonal navigation, interact genetically with engrailed to form posterior commissures in the developing VNC. The regulation of frazzled expression in engrailed-expressing neuroblasts contributes significantly to the formation of the posterior commissures by acting on axon growth. Finally, we identified a small genomic fragment within intron 1 of frazzled that can mediate activation by Engrailed in vivo when fused to a GFP reporter. These results indicate that Engrailed's function during the segregation of the neuroblasts is crucial for regulating different actors that are later involved in axon guidance. PMID- 17126318 TI - Activators of potassium M currents have anticonvulsant actions in two rat models of encephalitis. AB - Opioid systems in hippocampus regulate excitability and kappa opioids have a role in anticonvulsant protection, but their mechanisms of action are incompletely understood. We examined the ability of opioid and nonopioid agents with overlapping ionic mechanisms and actions similar to kappa opioid agonists, to block seizures in rat models of encephalitis due to Borna Disease virus and Herpes Simplex Virus Type-1. Naltrindole, a delta antagonist and thus a kappa opioid sparing agent, (10 mg/kg s.c.) blocked spontaneous and naloxone (opioid antagonist)-induced seizures in the models, but produced somatic signs similar to opioid withdrawal. Given that delta antagonists as well as kappa opioid agonists in hippocampus enhance potassium M currents (I(M)), we tested the effect of the I(M) augmenter flupirtine. Flupirtine (20 mg/kg i.p.) prevented seizures in Borna and herpes infected rats, without signs of withdrawal, hypotonia or sedation. The results support the efficacy of opioid and nonopioid drugs in modulating naloxone induced seizures in critical illness due to viral encephalitis and by analogy, opioid withdrawal seizures. PMID- 17126319 TI - Adenosine A1 receptors determine effects of caffeine on total fluid intake but not caffeine appetite. AB - Adenosine A1 receptor wild-type (+/+) and knockout (-/-) mice were used to elucidate the role of adenosine A1 receptors in caffeine self-administration in a two-bottle choice test and in the effect of caffeine on total fluid intake and plasma renin concentration. With access to water only, adenosine A1 receptor -/- mice showed greater basal fluid intake and greater plasma renin concentration than +/+ mice. Free access to both water and a caffeinated solution (30 mg/100 ml) for 14 days increased total fluid intake only in adenosine A1 receptor +/+ mice (by 23+/-3%), and both total fluid intake and plasma renin concentration were no longer different between genotypes. Mean intake of water and caffeinated solution was not different between adenosine A1 receptor +/+ and -/- mice. These data reveal that adenosine A1 receptors do not contribute to caffeine consumption, but determine the effects of caffeine on fluid intake and plasma renin concentration. PMID- 17126320 TI - Mechanisms underlying ATP-induced endothelium-dependent contractions in the SHR aorta. AB - In mature spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats, acetylcholine, the calcium ionophore A 23187 and ATP release endothelium-derived contracting factor (EDCF), cyclooxygenase (COX) derivatives that activate thromboxane-endoperoxide (TP) receptors on vascular smooth muscle. The EDCFs released by acetylcholine have been identified as prostacyclin and prostaglandin (PG) H(2) while in response to A 23187 thromboxane A(2), along with the two other prostaglandins, contributes to the endothelium-dependent contractions. The purpose of the present study was to identify the EDCFs produced by ATP. Isometric tension and the release of prostaglandins were measured in isolated aortic rings of WKY rats and SHR. ATP produced the endothelium-dependent release of prostacyclin, thromboxane A(2) and PGE(2) (PGI(2)>>TXA(2)> or =PGE(2)>PGF(2alpha)) in a similar manner in aorta from WKY rats and SHR. In SHR aortas, the release of thromboxane A(2) was significantly larger in response to ATP than to acetylcholine while that to prostacyclin was significantly smaller. The inhibition of cyclooxygenase with indomethacin prevented the release of prostaglandins and the occurrence of endothelium-dependent contractions. The thromboxane synthase inhibitor dazoxiben selectively abolished the ATP-dependent production of thromboxane A(2) and partially inhibited the corresponding endothelium-dependent contractions. U 51605, a non-selective inhibitor of PGI synthase, reduced the release of prostacyclin elicited by ATP but induced a parallel increase in the production of PGE(2) and PGF(2alpha), suggestive of a PGH(2)-spillover, which was associated with the enhancement of the endothelium dependent contractions. Thus, in the aorta of SHR, endothelium-dependent contractions elicited by ATP involve the release of thromboxane A(2) and prostacyclin with a possible contribution of PGH(2). PMID- 17126321 TI - kappa-opioid receptor stimulation inhibits cardiac hypertrophy induced by beta1 adrenoceptor stimulation in the rat. AB - To test the hypothesis that kappa-opioid receptor stimulation inhibits cardiac hypertrophy induced by beta1-adrenoceptor stimulation, we determined the effects of trans-(+/-)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl] benzeneacetamide methanesulfonate salt (U50,488H), a selective kappa-opioid receptor agonist, on cardiac hypertrophy induced by isoprenaline, a selective beta-adrenoceptor agonist, in neonatal ventricular myocytes upon blockade of beta2-adrenoceptor. Hypertrophy of cardiomyocytes was determined by increases in (i) total protein content; (ii) [3H]leucine incorporation; and iii) cell size. 10 micromol/l isoprenaline increased all three parameters. The effects were abolished by 2 micromol/l propranolol, a beta-adrenergic receptor antagonist, or 300 nmol/l CGP20712A, a beta1-adrenoceptor antagonist, but not by 100 nmol/l ICI118,551, a beta2-adrenoceptor antagonist. The effects were also abolished by Rp-cAMPs 100 micromol/l, a protein kinase A inhibitor and not by pertussis toxin 5 mg/l. The effects of isoprenaline in the presence or absence of ICI118,551 were also abolished by 1 micromol/l U50,488H. The inhibitory effects of U50,488H were abolished by 1 micromol/l nor-binaltorphimine, a selective kappa-opioid receptor antagonist. U50,488H also abolished the increases in the amplitude and frequency of the spontaneous intracellular Ca2+ transient induced by 10 micromol/l isoprenaline in the presence or absence of ICI118,551, an effect also abolished by nor-binaltorphimine. In conclusion the results show that kappa-opioid receptor stimulation abolished both the cardiac hypertrophy and enhanced amplitude and frequency of the spontaneous intracellular Ca2+ transient induced by beta1 adrenoceptor stimulation. PMID- 17126322 TI - [3H]HEMADO--a novel tritiated agonist selective for the human adenosine A3 receptor. AB - Adenosine A(3) receptors are promising drug targets for a number of conditions like inflammatory diseases including asthma, ischemic injury or certain types of cancer. Consequently, intense efforts are dedicated to the development of selective A(3) agonists and antagonists. The only tritiated agonist that is available for radioligand binding is the nonselective [(3)H]5'-N ethylcarboxamidoadenosine ([(3)H]NECA). Based on a recently characterized series of 2-substituted adenosine receptor agonists we developed a tritiated selective A(3) radioligand with high affinity. From this series 2-hexyn-1-yl-N(6) methyladenosine (HEMADO) with a K(i)-value of 1.1 nM at the human A(3) subtype was chosen. HEMADO is 300-fold selective versus the A(1) subtype, and 1100-fold and more than 25,000-fold selective compared to the adenosine A(2A) and A(2B) receptors, respectively. The tritiated derivative [(3)H]HEMADO exhibited the same affinity as the unlabeled precursor. In concentrations up to 10 nM no specific binding to adenosine A(1), A(2A) or A(2B) receptors was observed confirming the high selectivity for adenosine A(3) receptors. Characterization of [(3)H]HEMADO in radioligand binding studies revealed reversible binding to the human adenosine A(3) subtype. In saturation binding studies for the A(3) subtype a K(D)-value of 1.1 nM was determined. Nonspecific binding at a radioligand concentration of 1 nM amounted to 1-2% of total binding. Competition binding with a panel of adenosine receptor ligands clearly confirmed the correct A(3) pharmacology of the binding site labeled by [(3)H]HEMADO. With [(3)H]HEMADO we present a tritiated agonist with high affinity and A(3)-selectivity and very low nonspecific binding. [(3)H]HEMADO is a useful tool for specific screening for A(3) receptor agonists and antagonists in improved radioligand binding assays with the human subtype. PMID- 17126323 TI - Lipopolysaccharide-binding and neutralizing activities of surfactin C in experimental models of septic shock. AB - To evaluate the anti-endotoxin activity of surfactin C, we studied its lipopolysaccharide-binding activity in vitro and therapeutic efficacy in experimental models of gram-negative septic shock. The ability of surfactin C to bind LPS from Escherichia coli O111:B4 was determined using a limulus chromogenic assay. Male ICR mice and Sprague-Dawley rats were given intraperitoneal administration of 1x10(9) colony forming units of E. coli ATCC 25922. After bacterial challenge, all animals were randomized to receive intraperitoneally saline, polymyxin B or surfactin C. Surfactin C not only completely bound to the LPS (its median effective concentration being 13.75 microM) but also improved the survival and reduced of the number of inoculated bacteria in the mouse model of septic shock. Surfactin C reduced the plasma endotoxin, tumor necrosis factor alpha and nitric oxide levels in response to septic shock in rats. PMID- 17126324 TI - Leishmania mexicana: identification and characterization of an aspartyl proteinase activity. AB - An aspartyl proteinase activity was detected in the soluble fraction (SF) of Leishmania mexicana promastigotes by the use of the synthetic substrate benzoyl Arg-Gly-Phe-Phe-Leu-4-methoxy-beta-naphthylamide selective for Cathepsin D like aspartyl-proteinases. This peptide was hydrolyzed with an apparent K(m) of 2.3+/ 0.3 microM. The classic inhibitor of aspartyl-proteinases, diazo-acetyl norleucinemethylester (DAN) inhibited the proteolytic activity with an IC(50) of 400 microM. The soluble fraction degraded (in absence of thiol groups) human fibrinogen with a specific activity of 533 U/mg protein. When tested for the ability to inhibit the "in vitro" proliferation of L. mexicana promastigotes, DAN showed concentration dependent anti-proliferative effects with a LD(50) of 466 microM at 48 h, with a significant fall in this value to 22 microM after 72 h. This is the first characterization of an aspartyl-proteinase activity in Leishmania, calling for further studies directed towards the physiologic role of these enzymes in the parasite. The anti-proliferative effect of its inhibition makes this enzyme a putative new target for the development of leishmanicidal drugs. PMID- 17126325 TI - Pyrrolysine analogues as substrates for pyrrolysyl-tRNA synthetase. AB - In certain methanogenic archaea a new amino acid, pyrrolysine (Pyl), is inserted at in-frame UAG codons in the mRNAs of some methyltransferases. Pyl is directly acylated onto a suppressor tRNA(Pyl) by pyrrolysyl-tRNA synthetase (PylRS). Due to the lack of a readily available Pyl source, we looked for structural analogues that could be aminoacylated by PylRS onto tRNA(Pyl). We report here the in vitro aminoacylation of tRNA(Pyl) by PylRS with two Pyl analogues: N-epsilon-d-prolyl-l lysine (d-prolyl-lysine) and N-epsilon-cyclopentyloxycarbonyl-l-lysine (Cyc). Escherichia coli, transformed with the tRNA(Pyl) and PylRS genes, suppressed a lacZ amber mutant dependent on the presence of d-prolyl-lysine or Cyc in the medium, implying that the E. coli translation machinery is able to use Cyc tRNA(Pyl) and d-prolyl-lysine-tRNA(Pyl) as substrates during protein synthesis. Furthermore, the formation of active beta-galactosidase shows that a specialized mRNA motif is not essential for stop-codon recoding, unlike for selenocysteine incorporation. PMID- 17126326 TI - Extracellular heat shock protein 70 mediates heat stress-induced epidermal growth factor receptor transactivation in A431 carcinoma cells. AB - The initial steps of heat stress in A431 cells were previously characterized by ligand-independent EGFR transactivation via an unknown mechanism and concomitant secretion of Hsp70. In this work we demonstrate that the depletion of Hsp70 from the conditioned medium of heated cells abolishes EGFR transactivation indicating that secreted Hsp70 is essential for EGFR transactivation during heat shock. This notion is supported by the findings that purified Hsp70 can induce EGFR transactivation and the activation of EGFR-dependent signaling pathways. Both heat stress and pure Hsp70 stimulate activation of TLR2/4 and their association with EGFR. These results suggest that the secreted Hsp70 mediates the cross communication of TLR and EGFR signaling systems in A431 cells. PMID- 17126327 TI - Detection of nucleotide binding to Na,K-ATPase in an aqueous membrane suspension by 13C cross-polarization magic-angle spinning NMR spectroscopy. AB - Binding of uniformly (13)C labelled ATP to Na,K-ATPase was studied by (13)C cross polarization magic-angle spinning (CP-MAS) NMR. In the presence of 30 mM Na(+) , and with sample- and time-averaging, NMR spectra obtained at 4 degrees C exhibited several resonances for the bound nucleotide. Chemical shifts suggested that site-specific changes in the micro-environment or conformation of the nucleotide occurred in the high affinity binding site. These experiments permit further studies of nucleotide dynamics, structure and binding under physiologically relevant conditions. PMID- 17126328 TI - Two conserved domains in PCIF1 mediate interaction with pancreatic transcription factor PDX-1. AB - PCIF1 is a TRAF and POZ domain containing nuclear factor that interacts with and inhibits transactivation of pancreatic homeodomain transcription factor PDX-1. Here, we demonstrate interaction of endogenous PDX-1 and PCIF1 in MIN6 insulinoma cells. Within PCIF1, the TRAF and POZ domains are both required for physical and functional interaction with the C-terminus of PDX-1, whereas the C-terminal domain of PCIF1 directs its nuclear localization. A human PDX-1 mutation associated with diabetes, E224K, disrupts the ability of PCIF1 to inhibit PDX-1 transactivation, suggesting that the interaction between PDX-1 and PCIF1 is required for normal glucose homeostasis. Inhibition of transactivation occurs by a mechanism distinct from the classical role of POZ domains to recruit co repressors and histone deacetylases. Understanding the functional roles of PCIF1 domains may have application to therapeutic beta-cell replacement strategies involving PDX-1 for the treatment of diabetes. PMID- 17126330 TI - The cell membrane-shielding function of eicosapentaenoic acid for Escherichia coli against exogenously added hydrogen peroxide. AB - The colony-forming ability of catalase-deficient Escherichia coli mutant genetically modified to produce eicosapentaenoic acid (EPA) showed less decrease than in a control strain producing no EPA, when treated with 0.3mM hydrogen peroxide (H(2)O(2)) under non-growth conditions. H(2)O(2)-induced protein carbonylation was enhanced in cells lacking EPA. The amount of fatty acids was decreased more significantly for cells lacking EPA than for those producing EPA. Much lower intracellular concentrations of H(2)O(2) were detected for cells with EPA than those lacking EPA. These results suggest that cellular EPA can directly protect cells against oxidative damage by shielding the entry of exogenously added H(2)O(2). PMID- 17126329 TI - Comparative efficacy of HgCl2 with candidate aquaporin-1 inhibitors DMSO, gold, TEA+ and acetazolamide. AB - Aquaporin-1 (AQP1) inhibitors are predicted to have multiple clinical applications. Hg(++) is a non-specific and toxic AQP1 blocker. We compared compounds with reported AQP1 inhibition activity, including DMSO, Au(+++), Ag(+), tetraethylammonium and acetazolamide. Water permeability was measured by stopped flow light scattering in erythrocytes and volume marker dilution in epithelial cells. Au(+++) inhibited AQP1 with IC50 approximately 14 microM, similar to 10 microM for Hg(++). DMSO slowed osmotic equilibration; however, the apparent inhibition was due to 'osmotic clamp' rather than AQP1 inhibition. Neither tetraethylammonium nor acetazolamide (to 10 mM) inhibited AQP1. Our data indicate the need to identify new AQP1 inhibitors. PMID- 17126331 TI - Divergent evolutionary lines of fungal cytochrome c peroxidases belonging to the superfamily of bacterial, fungal and plant heme peroxidases. AB - Novel open reading frames coding for cytochrome c peroxidase (CcP) belonging to the superfamily of bacterial, fungal, and plant heme peroxidases were analyzed in the available fungal genomes. Multiple sequence alignment of 71 selected peroxidase genes revealed the presence of three conserved regions essential for their function: one on the distal and two on the proximal side of the prosthetic heme group. Conserved sequence motifs on the proximal heme side are peculiar for CcPs and are responsible for their reactivity. Phylogenetic analysis performed with the distance method as well as with the maximum likelihood method revealed the existence of three distinct subfamilies of fungal CcP and their relationship to other members of the peroxidase superfamily. These divergent CcP evolutionary lines apparently evolved from a single primordial heme peroxidase gene in parallel with the evolution of ascorbate peroxidase genes. Analyzed CcPs differ significantly in their N-terminal sequences. Only subfamily I did not exhibit a presence of any signal sequence. Subfamily II members possess a well defined signal sequence allowing processing and release into mitochondrion and also in subfamily III a signal sequence was detected. Several here analyzed peroxidase genes mainly from Candida albicans and from Rhizopus oryzae can be considered interesting for the investigation of the structure-function relationship of novel CcPs revealing differences to the well documented properties of cytochrome c peroxidase from Saccharomyces cerevisiae. PMID- 17126332 TI - Time-resolved responses to chemoattractant, characteristic of the front and tail of Dictyostelium cells. AB - In a gradient of chemoattractant, Dictyostelium cells are orientated with their front directed toward the source and their tail pointing into the opposite direction. The front region is specified by the polymerization of actin and the tail by the recruitment of filamentous myosin-II. We have dissected these front and tail responses by exposing cells to an upshift of cyclic AMP. A sharp rise and fall of polymerized actin within 10s is accompanied by the recruitment of proteins involved in turning actin polymerization on or off. The cortical accumulation of myosin-II starts when the front response has declined, supporting the concept of divergent signal transmission and adaptation pathways. PMID- 17126333 TI - Deep senescent human fibroblasts show diminished DNA damage foci but retain checkpoint capacity to oxidative stress. AB - Critically shortened telomeres trigger a DNA damage response in replicatively senescent cells. Here we report that while DNA damage foci can be detected in newly senescent cells, these foci eventually diminished in deep senescent cells. However, DNA checkpoint signalling and repair machinery in response to oxidative stress remain uncompromised in these deep senescent cells. Activation of p53 by oxidative stress is unaffected despite a marked decrease in expression of platelet-derived growth factor alpha-receptor. These findings suggest that cellular senescence is not a static process hence care must be taken in the selection of biomarkers of senescence in studies of ageing. PMID- 17126334 TI - Distinct sets of developmentally regulated genes that are expressed by human oocytes and human embryonic stem cells. AB - OBJECTIVE: To identify genes that are expressed differently during final oocyte maturation and early embryonic development in humans. DESIGN: Comparison of gene expression profiles of human germinal vesicle oocytes (hGVO), human embryonic stem cells (hESC) and human foreskin fibroblasts. SETTING: Research centers and a fertility unit in a university hospital. PATIENT(S): Fifty-five healthy women donated 76 hGVO. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Gene expression profiles were analyzed and compared with the use of microarray and reverse transcription polymerase chain reaction. RESULT(S): Altogether, 10,183 genes were expressed in hGVO, and 45% of these genes were unclassified by biologic function. Four oocyte-specific genes (MATER, ZAR1, NPM2 and FIGLA) were detected in hGVO for the first time. Known components of 4 signaling pathways (MOS-MPF, transforming growth factor-beta, WNT, and NOTCH) were also found expressed in hGVO, with some components detected in hGVO for the first time. Distinct sets of genes that were revealed by comparison of expression profiles between hGVO, hESC, and human foreskin fibroblasts appear to be involved in oocyte maturation and early embryonic development. CONCLUSION(S): We obtained, for the first time, a large amount of information on gene expression of hGVO as compared with hESC. These data, from a unique research material-human oocytes, can now be used to understand the molecular mechanisms of early human development. PMID- 17126335 TI - Clinical evaluation of endometriosis and differential response to surgical therapy with and without application of Oxiplex/AP* adhesion barrier gel. AB - OBJECTIVE: To correlate parameters of endometriosis obtained during routine clinical evaluation with the subsequent formation of adhesions following surgical treatment by laparoscopy. DESIGN: Randomized, controlled, double-blind, clinical trial. SETTING: Tertiary referral centers for the treatment of endometriosis. PATIENT(S): Thirty-seven patients (65 with adnexa) with stage I-III endometriosis; endometrioma-only patients were excluded. INTERVENTION: Laparoscopic surgical treatment of endometriosis, followed by randomization to Oxiplex/AP (FzioMed, Inc., San Luis Obispo, California) gel treatment (treated group) of adnexa, or surgery alone (control group); follow-up laparoscopy 6-10 weeks later. MAIN OUTCOME MEASURE(S): Adnexal Americn Fertility Society score, correlated with color and location of endometriosis, as well as stage of disease determined by masked review of videotapes. RESULT(S): Control patients with at least 50% red lesions had a greater increase in ipsilateral adnexal adhesion scores than patients with mostly black or white and/or clear lesions. Treated patients with red lesions had a greater decrease in adnexal adhesion scores than control patients. There was a correlation between baseline endometriosis stage and postoperative adhesion formation in control patients, but not treated patients. CONCLUSION(S): Patients with red endometriotic lesions had greater increases in their adhesion scores than patients with only black, white, and/or clear lesions. Oxiplex/AP gel was effective in reducing adhesions, compared to surgery alone, in all groups. PMID- 17126336 TI - Sperm chromosome analysis in two cases of paracentric inversion. AB - OBJECTIVE: To examine sperm meiotic segregation in men with paracentric inversions. DESIGN: Cases reports, literature review. SETTING: Departments of reproductive biology, cytogenetics, gynaecology, and obstetrics. PATIENT(S): Two patients referred for infertility, heterozygous for a paracentric inversion. INTERVENTION(S): Fluorescence in situ hybridization (FISH) with specific probes and X/Y/18 centromeric probes on 1,000 spermatozoa for the 2 patients and 10 controls. MAIN OUTCOME MEASURE(S): Sperm aneuploidy frequency. RESULT(S): The FISH analysis using the specific probes for the paracentric inversion indicated low disequilibrium (0.4% and 0.5%). The FISH analysis using X/Y/18 centromeric probes indicated aneuploidy frequencies (0.3% and 1.1%), identical to those of control patients with the same sperm parameters. CONCLUSION(S): Paracentric inversion seems to be associated with a very low risk of aneuploidy. A larger study is necessary to explore all chromosome inversions. PMID- 17126337 TI - Uteroplacental circulation in early pregnancy complicated by threatened abortion supplemented with vaginal micronized progesterone or oral dydrogesterone. AB - OBJECTIVE: To compare the influence of vaginal micronized progesterone and oral dydrogesterone supplementation on uteroplacental circulation in early pregnancy that is complicated by threatened abortion. DESIGN: Randomized, parallel group, double-blind, double dummy-controlled study. SETTING: Tertiary care university hospital. PATIENT(S): Fifty-three patients with threatened abortion and a living embryo. INTERVENTION(S): Three hundred milligrams of micronized vaginal progesterone or 30 mg of oral dydrogesterone daily supplementation for 6 weeks, serial transvaginal Doppler ultrasound measurement of pulsatility index, resistance index, and systolic/diastolic ratio of the spiral arteries, the uterine arteries, and the intrachorionic area. MAIN OUTCOME MEASURE(S): Uteroplacental blood flow. RESULT(S): The study demonstrated that vaginal progesterone administration, but not oral dydrogesterone treatment, results in the decrease in the spiral artery pulsatility and resistance index and systolic/diastolic ratio. Insignificant decrease in pulsatility index and resistance index of the uterine artery was observed at >9 weeks and was not associated with treatment regimen. Dydrogesterone treatment was only accompanied by the decrease in the uterine artery systolic/diastolic ratio. CONCLUSION(S): Vaginal progesterone and oral dydrogesterone supplementation have a different influence on the uteroplacental circulation in early pregnancy that is complicated by threatened abortion. PMID- 17126338 TI - Female genital malformations and their associated abnormalities. AB - OBJECTIVE: With an incidence of up to 5% in the general population, genital malformations are a frequent clinical occurrence. The aim of this study was to assess whether a connection could be demonstrated between various degrees of severity of genital malformations and associated abnormalities. DESIGN: All patients were classified using the Vagina, Cervix, Uterus, Adnex, and Associated Malformation (VCUAM) classification. SETTING: University hospital. PATIENT(S): Two hundred eleven premenopausal patients with female genital malformations. INTERVENTION(S): The patients underwent diagnostic workup for genital malformations using laparoscopy as well as hysteroscopy. Associated malformations were detected by either magnetic resonance imaging (MRI) or ultrasound. MAIN OUTCOME MEASURE(S): Demonstration of a connection between various degrees of severity of genital malformations and associated abnormalities. RESULT(S): In 72 cases (36%) out of 202 patients with uterine malformations (VCUAM U1-4) we found associated abnormalities. The predominant findings were alterations in the renal system. When vaginal abnormality (VCUAM V1-5) alone was taken into consideration, an associated developmental disturbance in the renal tract was found in 30% of cases (n = 32 from 107). CONCLUSION(S): A close connection was demonstrated between genital malformations and associated abnormalities. For this reason, the diagnostic workup in patients with malformations should always include the renal system. Depending on the severity of the clinical picture, examinations may need to be extended further. PMID- 17126339 TI - Lower apoptosis rate in human cumulus cells after administration of recombinant luteinizing hormone to women undergoing ovarian stimulation for in vitro fertilization procedures. AB - OBJECTIVE: To investigate the effects of recombinant (r-) LH supplementation in "low responder" patients undergoing ovarian stimulation with r-FSH for an IVF program. The apoptosis rate in cumulus cells was used as an indicator of oocyte quality. DESIGN: Comparison of the rate of DNA fragmentation and caspase-3 activity in cumulus cells in women stimulated with r-LH and r-FSH, versus patients treated with r-FSH alone (control). SETTING: In vitro fertilization (IVF) laboratory. PATIENT(S): Forty patients undergoing assisted fertilization programs treated with a GnRH agonist, or r-FSH treatment begun on day 3 of the cycle (control). In the r-LH group, from day 8 of gonadotropin stimulation, 150 IU per day of r-LH were administered. INTERVENTION(S): Terminal deoxynucleotidyl transferase-mediated digoxigenin-deoxyuridine-triphosphate (dUTP) nick-end labeling (TUNEL) assay, and anti-caspase-3 cleaved immunoassay, to detect apoptosis in human cumulus cells. MAIN OUTCOME MEASURE(S): Difference in DNA fragmentation rate between cumulus cells derived from r-LH treatment and cumulus cells derived from control patients. RESULT(S): No differences were observed between the two groups in the total amount of r-FSH administered and in the number of retrieved oocytes per patient. A statistically significant increase in the number of immature oocytes and in the E(2) serum peak was observed in the control group. The number of transferred embryos was significantly higher in the r-LH group. Pregnancy and implantation rates were higher in the r-LH group, but without statistical significance. The apoptosis rate in cumulus cells was higher in the control group than in the r-LH group. CONCLUSION(S): This study suggests that supplementation with r-LH improves the chromatin quality of cumulus cells involved in the control of oocyte maturation. PMID- 17126340 TI - The expression of CD9 in the peri-implantation mouse uterus is upregulated in an ovarian steroid hormone-dependent manner. AB - OBJECTIVE: To examine the spatiotemporal expression of CD9 protein in the peri implantation mouse uterus as well as the effects of ovarian steroid hormones on CD9. DESIGN: Experimental animal study. SETTING: Reproductive biological center of Chinese Academy of Sciences. ANIMAL(S): Female Kunming white strain mice (6-8 weeks old). INTERVENTION(S): Subcutaneous injection of P(4)/E(2); uterine tissues were collected at different times after injection. MAIN OUTCOME MEASURE(S): The levels of protein and mRNA were detected in mouse uterus during peri-implantation and after steroid hormones treatment. RESULT(S): CD9 protein was expressed intensely in the stromal cells on days 1 and 2 of pregnancy. On days 3 and 4, the glandular and luminal epithelial cells exhibited accumulation of CD9 protein. After the initial attachment reaction on day 5, luminal epithelial and stromal cells immediately surrounding the blastocysts exhibited distinct accumulation of CD9. On days 6-8, the accumulation of CD9 occurred in decidual cells. Using ovariectomized mice, we also observed that both progesterone and estrogen upregulated uterine CD9 expression. CONCLUSION(S): The results of the current investigation showed that CD9 was differentially expressed in the uterus depending on the stage of implantation and was upregulated in ovarian steroid hormone-dependent manner, implicating multiple roles of CD9 in the regulation of embryo implantation during the peri-implantation period. PMID- 17126341 TI - Bowel urgency in patients with irritable bowel syndrome. AB - BACKGROUND & AIMS: Bowel urgency is the most bothersome symptom in irritable bowel syndrome patients with diarrhea, but its pathophysiology is poorly understood. Our aim was to assess the relationships among reporting the symptom, the reservoir functions of the colon and rectum, and the patients' psychologic profile. METHODS: The study involved 28 consecutive patients with irritable bowel syndrome and 17 healthy subjects. The presence or absence of bowel urgency was verified by means of a questionnaire during the 3 days required for the ingestion of radio-opaque markers. On the fourth day, an abdominal x-ray was taken to assess colonic transit time, and rectal sensory and motor responses were measured during rectal distention. The subjects' psychologic profiles were assessed using a psychologic symptoms checklist. RESULTS: Forty-six percent of the patients reported urgency associated with at least 1 defecation. The multivariate logistic regression analysis showed that colonic transit was the only variable independently associated with reported bowel urgency, but the threshold for the sensation of urgency was not removed from the model since its borderline significance level. Rectal compliance was closely associated with the threshold for the sensation of urgency during rectal distention but was not an independent factor for reporting the sensation. The patients with and without urgency showed altered psychologic profiles. CONCLUSIONS: The symptom of urgency is associated with objective alterations in the colonic and rectal reservoir of patients with irritable bowel syndrome. PMID- 17126342 TI - C-terminal apolipoprotein E-derived peptide, Ep1.B, displays anti-atherogenic activity. AB - OBJECTIVE: Apolipoprotein E (ApoE) is a lipid transport protein with expanded functions in cellular responses to tissue injury, immune regulation and cell growth. ApoE directs vascular changes that contribute to arterial protection as evidenced by the fact that isoforms of ApoE and ApoE deficiency correlate closely with accelerated plaque growth. The N-terminus of the ApoE protein has well characterized functions, displaying lipid-binding and anti-atherogenic activity, whereas the function of the C-terminus is only partially defined. We have assessed the effects of a 14 amino acid C-terminal ApoE peptide, termed Ep1.B (239-252), on intimal neoplasia in animal models. This peptide is a fragment of a naturally processed peptide (236-252) of murine ApoE. METHODS AND RESULTS: Ep1.B injection reduced neointimal hyperplasia after vascular surgery in rats and mice. When given during early plaque progression in ApoE-deficient mice, Ep1.B injections also prevented plaque growth. Treatment with Ep1.B did not, however, reduce established plaque growth in older mice. Peptides with alanine substitution of amino acid 249, Ep1.N, and with complete sequence reversal, Ep1.R, did not consistently inhibit plaque growth. CONCLUSION: A naturally processed C-terminal ApoE peptide, Ep1.B, has anti-atherogenic activity indicating a role for this naturally metabolized peptide in vascular wound healing and lipid homeostasis. PMID- 17126343 TI - Serum C-reactive protein response to percutaneous coronary intervention in patients with unstable or stable angina pectoris is associated with the risk of clinical restenosis. AB - BACKGROUND: Inflammation plays a pivotal role in the pathogenesis of atherosclerosis. Previous reports have used vaccination as a model to stimulate inflammation. The aim of the present study was to investigate the role of C reactive protein response to PCI in the risk of clinical restenosis or new coronary stenosis, considering PCI as a model to stimulate inflammation. MATERIAL AND METHODS: Eight hundred and ninety-one patients with stable or unstable angina pectoris and with normal serum troponin T1.0 cm or placental separation were considered abnormal. Placentitis was confirmed by histology of fetal membranes. Control mares had normal progestin profiles, transrectal ultrasonographic and clinical examinations. Control foals were born after Day 329; six were viable and one died after dystocia. All inoculated mares developed placentitis and foaled before Day 314. Thirteen of 15 foals were not viable. All inoculated mares had abnormal progestin profiles and 13 of the 15 were identified by the abbreviated progestin profile. Transrectal CTUP was affected by gestational age and increased after inoculation (P<0.05). Nine of 15 inoculated mares had a CTUP>1.0 cm by 5-day post-inoculation. By performing both tests, 20 of 22 mares were correctly identified with respect to pregnancy outcome. However, three inoculated mares exhibited minimal clinical signs and likely would not be examined in a clinical setting. These tests were diagnostic for identifying feto-placental compromise in the mare. PMID- 17126393 TI - The effect of five silane coupling agents on the bond strength of a luting cement to a silica-coated titanium. AB - OBJECTIVES: The adhesive performance of five silane coupling agents in adhering resin composite cement (3M ESPE) to silica-coated titanium was evaluated. Titanium was tribochemically silica-coated by using the Rocatec system. METHODS: Two volume percent solutions of 3-acryloyloxypropyltrimethoxysilane (Toray Dow Corning Silicone), N-[3-(trimethoxysilyl)propylethylenediamine] (Dow Corning), 3 mercaptopropyltrimethoxysilane (Toray Dow Corning Silicone) and bis-[3 (triethoxysilyl)propyl]polysulfide (Dow Corning) were prepared in 95 vol.% acidified ethanol and allowed to activate (hydrolyze). A pre-activated ca. 2 vol.% 3-methacryloyloxypropyltrimethoxysilane (ESPE Sil) was used as a control. The silanes were applied onto silica-coated titanium slides. Chemical activation reactions of the silanes were monitored by Fourier transform infrared spectrometry (Perkin-Elmer Spectrum One). RelyX ARC (3M ESPE) resin composite cement stubs were applied and photo-polymerized onto silica-coated titanium. The specimens were thermo-cycled (6000 cycles, 5-55 degrees C). RESULTS: Statistical analysis (ANOVA) showed that the highest shear bond strength (n=8 per group) value after thermocycling, 14.8 MPa (S.D. 3.8 MPa), was obtained with 2.0 vol.% 3 acryloyloxypropyltrimethoxysilane. Silanization and results with 3 methacryloyloxypropyltrimethoxysilane (control, ESPE Sil) did not statistically differ from 3-acryloyloxypropyltrimethoxysilane, 14.2 MPa (S.D. 5.8). The lowest shear bond strength was 7.5 (S.D. 1.9) MPa for N-[3 (trimethoxysilyl)propylethylenediamine] and 7.5 (S.D. 2.5) MPa for bis-[3 (triethoxysilyl)propyl]polysulfide. Both the type of silane (p<0.001) and storage conditions affected significantly the shear bond strength values (p<0.001). All silanes became activated according to the infrared spectroscopic analysis. SIGNIFICANCE: Silanization with 3-acryloyloxypropyltrimethoxysilane or 3 mercaptopropyltrimethoxysilane might offer an alternative for bonding a luting cement to silica-coated titanium. PMID- 17126394 TI - Probing molecular recognition sites on biosurfaces using AFM. AB - Knowledge of the molecular forces that drive receptor-ligand interactions is a key to gain a detailed understanding of cell adhesion events and to develop novel applications in biomaterials science. Until recently, there was no tool available for analyzing and mapping these forces on complex biosurfaces like cell surfaces. During the past decade, however, single-molecule atomic force microscopy (AFM) has opened exciting new opportunities for detecting and localizing molecular recognition forces on artificial biosurfaces and on living cells. In this review, we describe the general principles of the AFM technique, present procedures commonly used to prepare samples and tips, and discuss a number of applications that are relevant to the field of biomaterials. PMID- 17126395 TI - The construction of three-dimensional micro-fluidic scaffolds of biodegradable polymers by solvent vapor based bonding of micro-molded layers. AB - It is increasingly important to control cell growth into and within artificial scaffolds. Tissues such as skin, blood vessels, and cartilage have multi-layer structures with different cells in each layer. With the aid of micro-fabrication technology, a novel scaffolding method for biodegradable polymers such as polylactic acid (PLA), polyglycolic acid (PGA), and the copolymers poly(lactide co-glycolide)(PLGA), was developed to construct three-dimensional multi-layer micro-fluidic tissue scaffolds. The method emphasizes micro-fluidic interconnections between layers within the scaffolds and maintenance of high resolution geometries during the bonding process for the creation of multi layered scaffolds. Micro-holes (10-100 microm), micro-channels, and micro cavities were all created by micro-molding. Solvent-vapor based bonding of micro molded layers preserved 20 microm sized structures. Sample scaffolds were constructed for purposes such as channel-directed cell growth and size-based cell sorting. Further extension of these techniques to create a micro-vascular network within or between layers is possible. Culturing of human coronary artery endothelial cells (HCAECs) on the sample scaffolds demonstrated the biocompatibility of the developed process and the strong influence of high resolution micro-geometries on HCAEC growth. PMID- 17126396 TI - Vascular delivery of c-myc antisense from cationically modified phosphorylcholine coated stents. AB - c-Myc is involved in the formation of neointimal hyperplasia. We investigated in vitro, ex vivo and in vivo release of antisense c-myc from cationically modified phosphorylcholine-coated stents, as well as the effects on c-Myc expression and neointima formation in a porcine coronary stent model. In vitro experiments were performed to determine optimal loading of stents with antisense. Stents loaded with labelled antisense were deployed in porcine arteries ex vivo and in vivo. Antisense was detected in the vessel wall directly surrounding the stent of pig carotid and coronary artery up to 48 h after stent deployment. Nuclear uptake was observed in endothelial and vascular smooth muscle cells. Labelled antisense within peripheral tissues in vivo was <1.0% of that within stented arterial segments. Control and antisense loaded stents implanted into 10 pig coronary arteries and analysed at 28 days post-stenting showed that lumen area within the antisense stents was significantly increased (i.e. 30.5% greater, P<0.01), whilst both neointimal area and neointimal thickness were significantly reduced (17.5% and 19.5%, respectively, P<0.01) compared to control stents. Cationically modified phosphorylcholine coated stent-based delivery of c-myc antisense is feasible with minimal systemic delivery and is associated with a reduction of in stent neointimal hyperplasia in pig coronary arteries. PMID- 17126397 TI - Antibody arrays for quantitative immunophenotyping. AB - Detection of multiple surface antigens expressed on living cell is an important step for cell processing and clinical diagnosis. Here we describe the preparation of antibody arrays that allow parallel detection of multiple surface antigens through affinity binding of living cells. An antibody array was fabricated by photo-assisted patterning of an alkanethiol monolayer formed on a gold-coated glass plate and subsequent immobilization of antibodies specific for cell surface antigens in an array format. We demonstrate here that rapid phenotyping can be performed on the array for both adhesion-dependent and non-dependent cells by direct cell binding assays. The density of bound cells on each antibody spot was in accordance with their contents in an original suspension. This result suggests the feasibility of the array-based method for quantitative assessment of multiple antigen expression. These findings will serve to extend the range of fundamental and clinical applications of antibody arrays. PMID- 17126398 TI - A patient with de novo AML M1 and t(16;21) with karyotype evolution. PMID- 17126399 TI - Lymphoid organ development in rabbits: major lymphocyte subsets. AB - Although rabbits represent an important animal model, little is known about the lymphoid organ development in this species. In the present study, lymphocyte subsets in peripheral blood, spleen, mesenteric and popliteal lymph nodes in newborn and 2-, 4-, 6- and 8-week old and adult were characterized. Lymphocyte subsets were detected using flow cytometry and monoclonal antibodies against rabbit CD4, CD8, T-cell-specific antigen and cross-reactive antibody against B cell antigen CD79alpha. In neonates, lower numbers of T cells were detected in both peripheral blood and spleen than in mesenteric lymph nodes. In comparison with other compartments, CD79alpha(+) cells prevailed in the spleen. Post-natal development was characterized by a decreased CD4(+)/CD8(+) ratio due to increasing frequency of CD8(+) lymphocytes in all organs but mesenteric lymph nodes, where it was due to decreased numbers of CD4(+) lymphocytes. Another significant feature was the increase of B cells in peripheral blood and mesenteric lymph nodes. PMID- 17126400 TI - Channel catfish hepcidin expression in infection and anemia. AB - Hepcidin, originally identified as a 25 amino acid antimicrobial peptide made in the liver, is a key regulator of iron balance and recycling in humans and mice. Closely related hepcidin genes and peptides have also been identified in a number of fish species and in teleosts are thought to function as endogenous antibiotics involved in host defense against infection. Here we report the transcriptional regulation of hepcidin expression by infection and anemia in the channel catfish. Changes in hepcidin expression in catfish challenged with Edwardsiella ictaluri and in fish affected by channel catfish anemia (CCA) were measured by real time quantitative PCR. Hepcidin transcript levels in the livers were increased 4, 19, and 22-fold at 4, 24, and 48h following bacterial challenge, respectively. However, augmented hepcidin expression in the intestine and olfactory sac was detected only at 48h post-infection. Hepcidin transcript levels in the livers of catfish affected by CCA were less than 14% of that present in healthy counterparts. Hepatic hepcidin transcript levels correlated significantly with serum iron concentrations (r=0.54, p<0.05) and with the percent saturation of transferrin (r=0.63, p<0.05). Similar to mammalian hepcidins, channel catfish hepcidin is an iron-responsive gene and may also play important roles in innate host defense to infection and in iron homeostasis. Mammalian hepcidins may have evolved from an antimicrobial peptide and its structure and transcriptional regulatory mechanisms have been conserved throughout vertebrate evolution. PMID- 17126402 TI - Phosphatidyl inositol-3-phosphate kinase mediates CD14 dependent signaling. AB - Lipopolysaccharide (LPS), a component of the cell wall of Gram-negative bacteria, is an important mediator of innate immunity and septic shock, but the exact mechanisms mediating cellular LPS recognition and the subsequent translation to inflammatory gene expression remain incompletely understood. CD14 has been established as a receptor that confers high sensitivity to LPS in cells of the myeloid lineage, probably by presenting LPS to Toll receptors. We use an anti CD14 blocking antibody to define a LPS stimulus that activates only this high affinity component of the LPS receptor and then examine CD14 dependent signaling events that are activated in response to LPS stimulation. We describe a novel LPS activated signaling pathway in human PBMC that leads to cytokine production and is mediated by PI3 kinase through Ras and the MEK/ERK cassette. Moreover, we show the PI3 kinase effectors PKB and PKC(zeta) are also activated by PI3 kinase in a CD14 dependent manner in LPS stimulated human PBMC. Thus, PI3 kinase appears to be an essential component in LPS signal transduction. PMID- 17126403 TI - Central corneal thickness in the European Glaucoma Prevention Study. AB - PURPOSE: To measure central corneal thickness (CCT) within the participants of the European Glaucoma Prevention Study (EGPS). This study was designed to test if lowering intraocular pressure (IOP) by means of dorzolamide is able to prevent or delay conversion from ocular hypertension to glaucoma. DESIGN: Randomized, double masked, controlled, observational clinical trial. PARTICIPANTS: Eight hundred fifty-four of 1077 ocular hypertensive participants within the EGPS were investigated. Four hundred twenty-nine patients were treated with dorzolamide and 425 patients received placebo. INTERVENTION: Treatment with dorzolamide or placebo (the vehicle of dorzolamide) in 1 or both eyes. MAIN OUTCOME MEASURES: Central corneal thickness as measured by ultrasound pachymetry (DGH-500 Pachette; DGH Technologies, Exton, PA). The CCT measurements were obtained in the morning before measuring IOP. Five measurements were taken from each eye of each patient within 5 minutes of application of anesthetic eye drops. RESULTS: Mean CCT was 572.6+/-37.4 microm (range, 458.5-695.6 microm). The CCT was higher in younger patients, male patients, and diabetic patients. Mean CCTs for the 429 patients receiving dorzolamide were 574.2+/-38.48 microm (range, 458.5-695.6 microm) and 571.0+/-36.21 microm (469.7-690.1 microm) for the 425 patients receiving placebo (P = 0.205). Central corneal thickness did not correlate with refraction, baseline IOP, or systemic hypertension. CONCLUSION: Central corneal thickness measurements within the EGPS were greater than those reported in other studies of normal eyes without ocular hypertension. Larger CCT measurements correlated with male gender, younger age, and diabetes. PMID- 17126404 TI - The complex multidomain organization of SCO-spondin protein is highly conserved in mammals. AB - The multidomain organization of SCO-spondin protein is a special feature of the chordate phylum. This protein is expressed in the central nervous system (CNS) from the time a dorsal neural tube appears in the course of phylogenetical evolution. With the advance of the systematic whole genomes sequencing, we were able to determine the SCO-spondin amino acid sequence in four mammalian species using the Wise2 software. From the ClustalW alignment of bovine (Bos taurus), human (Homo sapiens), murine (Mus musculus) and rat (Rattus norvegicus) proteins, a consensus sequence for mammalian SCO-spondin was determined and further validated with the dog (Canis familiaris) SCO-spondin sequence. The analysis of this consensus sequence is consistent with a very high degree of conservation in the amino acids composition and multidomain organization of SCO-spondin in mammals. In addition, the identification of conserved domains, namely, Emilin (EMI), von Willebrand factor D (vWD), low-density lipoprotein receptor type A (LDLrA) domains, SCO repeats (SCOR), thrombospondin type 1 repeats (TSR), a coagulation factor 5/8 type C (FA5-8C) or discoidin motif and a C-terminal cystine knot (CTCK) domain, provides a greater insight into the putative function of this multidomain protein. SCO-spondin belongs to the TSR superfamily given the presence of a great number of TSR (26). A finer classification of the TSR motifs in groups 1, 2 and 3 is proposed on the basis of different cysteine patterns. Interestingly, group 2 TSR are present in a number of CNS developmental proteins including R-spondins, F-spondins and Mindins. PMID- 17126405 TI - Techniques of preparing plant material for chromatographic separation and analysis. AB - This paper discusses preparation techniques of samples of plant material for chromatographic analysis. Individual steps of the procedures used in sample preparation, including sample collection from the environment or from tissue cultures, drying, comminution, homogenization, leaching, extraction, distillation and condensation, analyte enrichment, and obtaining the final extracts for chromatographic analysis are discussed. The techniques most often used for isolation of analytes from homogenized plant material, i.e., Soxhlet extraction, ultrasonic solvent extraction (sonication), accelerated solvent extraction, microwave-assisted extraction, supercritical-fluid extraction, steam distillation, as well as membrane processes are emphasized. Sorptive methods of sample enrichment and removal of interferences, i.e., solid-phase extraction, and solid-phase micro-extraction are also discussed. PMID- 17126406 TI - Enhanced creativity in bipolar disorder patients: a controlled study. AB - OBJECTIVE: Associations between eminent creativity and bipolar disorders have been reported, but there are few data relating non-eminent creativity to bipolar disorders in clinical samples. We assessed non-eminent creativity in euthymic bipolar (BP) and unipolar major depressive disorder (MDD) patients, creative discipline controls (CC), and healthy controls (HC). METHODS: 49 BP, 25 MDD, 32 CC, and 47 HC (all euthymic) completed four creativity measures yielding six parameters: the Barron-Welsh Art Scale (BWAS-Total, and two subscales, BWAS Dislike and BWAS-Like), the Adjective Check List Creative Personality Scale (ACL CPS), and the Torrance Tests of Creative Thinking--Figural (TTCT-F) and Verbal (TTCT-V) versions. Mean scores on these instruments were compared across groups. RESULTS: BP and CC (but not MDD) compared to HC scored significantly higher on BWAS-Total (45% and 48% higher, respectively) and BWAS-Dislike (90% and 88% higher, respectively), but not on BWAS-Like. CC compared to MDD scored significantly higher (12% higher) on TTCT-F. For all other comparisons, creativity scores did not differ significantly between groups. CONCLUSIONS: We found BP and CC (but not MDD) had similarly enhanced creativity on the BWAS-Total (driven by an increase on the BWAS-Dislike) compared to HC. Further studies are needed to determine the mechanisms of enhanced creativity and how it relates to clinical (e.g. temperament, mood, and medication status) and preclinical (e.g. visual and affective processing substrates) parameters. PMID- 17126407 TI - Seasonality and suicide in Italy: amplitude is positively related to suicide rates. AB - BACKGROUND: Recent studies have reported changes in the time patterns of suicide, with conflicting findings regarding the direction of these changes: data from Italy were investigated to evaluate the influence of recent social and medicine related changes on the seasonality of suicides in the country. METHODS: A total of 71,227 male suicides and 26,466 female suicides occurring in Italy from 1974 to 2003 were investigated with harmonic spectral analysis to extract their monthly seasonal dispersion by five-year intervals. RESULTS: The suicide rates of both males and females showed a rising trend, with an evident peak in the 1987 1994 period and a decrease thereafter. Seasonality of suicides, with a clear peak in spring as against the other seasons, accounted for a statistically significant proportion of total variance: around 40% among males and 39% among females. Seasonality did not change across time in a relevant way; however, an anticipation of the peak was observed in both males and females over time, with amplitude increasing or decreasing as a function of yearly suicide rates. LIMITATIONS: Data could not be analysed according to age or to the method of suicide, since this information was not available across the whole time interval. CONCLUSION: The seasonal effect on mortality by suicide is positively related to suicide rates, so much that changes in suicide rates over time correspond to changes in suicide seasonality. PMID- 17126408 TI - Temperament-creativity relationships in mood disorder patients, healthy controls and highly creative individuals. AB - OBJECTIVE: To investigate temperament-creativity relationships in euthymic bipolar (BP) and unipolar major depressive (MDD) patients, creative discipline controls (CC), and healthy controls (HC). METHODS: 49 BP, 25 MDD, 32 CC, and 47 HC (all euthymic) completed three self-report temperament/personality measures: the Revised NEO Personality Inventory (NEO-PI-R), the Temperament Evaluation of the Memphis, Pisa, Paris, and San Diego Autoquestionnaire (TEMPS-A), and the Temperament and Character Inventory (TCI); and four creativity measures yielding six parameters: the Barron-Welsh Art Scale (BWAS-Total, BWAS-Like, and BWAS Dislike), the Adjective Check List Creative Personality Scale (ACL-CPS), and the Torrance Tests of Creative Thinking--Figural (TTCT-F) and Verbal (TTCT-V) versions. Factor analysis was used to consolidate the 16 subscales from the three temperament/personality measures, and the resulting factors were assessed in relationship to the creativity parameters. RESULTS: Five personality/temperament factors emerged. Two of these factors had prominent relationships with creativity measures. A Neuroticism/Cyclothymia/Dysthymia Factor, comprised mostly of NEO-PI R-Neuroticism and TEMPS-A-Cyclothymia and TEMPS-A-Dysthymia, was related to BWAS Total scores (r=0.36, p<0.0001) and BWAS-Dislike subscale scores (r=0.39, p<0.0001). An Openness Factor, comprised mostly of NEO-PI-R-Openness, was related to BWAS-Like subscale scores (r=0.28, p=0.0006), and to ACL-CPS scores (r=0.46, p<0.0001). No significant relationship was found between temperament/personality and TTCT-F and TTCT-V scores. CONCLUSIONS: Neuroticism/Cyclothymia/Dysthymia and Openness appear to have differential relationships with creativity. The former could provide affective (Neuroticism, i.e. access to negative affect, and Cyclothymia, i.e. changeability of affect) and the latter cognitive (flexibility) advantages to enhance creativity. Further studies are indicated to clarify mechanisms of creativity and its relationships to affective processes and bipolar disorders. PMID- 17126409 TI - Manifestations of affective disturbance in sub-Saharan Africa: key themes. AB - BACKGROUND: Affective disorder, once seen as rare in sub-Saharan Africa, is now viewed as more common. There are however challenges in assessing rates of depression, exploring manifestations of depression, and understanding risk factors for depression, especially those related to gender. AIM: To identify key themes and new directions for research on affective disorder in sub-Saharan Africa, with particular reference to gender issues. METHODS: Interpretive review of selected literature, and theoretical analysis. RESULTS: There are gender differences in manifestations and rates of depression in sub-Saharan Africa, and there are forms of presentation (largely somatic, based on interpersonal relationships, or spiritual in nature) which may obscure the detection of depression. Studies are consistent however that when depressive symptoms are sought and are present, these are reasonably easy to elicit. LIMITATIONS: This was not a systematic review, and the substantial grey literature from sub-Saharan Africa was not reviewed. For many countries, there are no data available. CONCLUSIONS: Affective disorder, when properly sought for through both qualitative and quantitative methods, has been found to be common in sub-Saharan Africa. There is a paucity of research on interventions with affective disorder in this region. PMID- 17126410 TI - Immunoblot analysis of IgG antibody response to Sarcoptes scabiei in swine. AB - This study was performed to determine the frequencies and specificities of IgG antibodies binding to component of Sarcoptes scabiei extracts in swine with hypersensitive and chronic mange. The hypersensitive form is characterised by pruritus and the presence of small red papules over the flanks and belly. The chronic form is characterised by crusts, which contain large numbers of mites and are attached to the skin; the lesions are most commonly found on the internal pinna extending into the auditory canal. S. scabiei mite extract was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with subsequent immunoblotting. IgG-binding proteins were detected with individual sera from 30 hypersensitive and 21 chronically infected pigs; eight "Specific Pathogen Free" pigs were used as negative controls. Seven protein bands with molecular weights ranging from >220 to 30 Kilodalton (KDD) (>220, 218, 110, 80, 66, 52, 36 KDD) strongly bound with IgG antibodies; five out of these seven components (218, 110, 80, 66, 52 KDD) bound also with sera from negative pigs. There is a statistically significant difference in the antigenic recognition spectra between hypersensitive and chronically infected pigs; component of >220 KDD is more frequently recognized by chronically infected pigs (P=0.0006, chi(2)=11.74), in contrast component of 36 KDD is more frequently recognized by hypersensitive pigs (P=0.001, chi(2)=10). Our results clearly indicate there is a difference in the reactivity to antigenic peptides/proteins of S. scabiei mite between hypersensitive and chronically infected pigs, and revealed that only two antigens may be considered S. scabiei-specific and used for diagnostic purposes in swine. PMID- 17126411 TI - Pharmacogenetics of MXA SNPs in interferon-beta treated multiple sclerosis patients. AB - The myxovirus resistance A (MXA) mRNA has been extensively investigated for assessing the biologic responses of multiple sclerosis (MS) patients to interferon-beta (IFN-beta) therapy. The objective of this study was to evaluate the associations between two MXA promoter region single nucleotide polymorphisms (rs2071430 and rs17000900) and the gene expression responses, clinical and MRI phenotypes in IFN-beta treated MS patients. The rs2071430 and rs17000900 SNPs, which are located in or near an interferon-stimulated response element (ISRE), were genotyped in 179 relapsing MS patients. Quantitative MRI measurements were available for 101 patients on IFN-beta monotherapy. Gene expression was assessed in 22 anti-interferon-beta neutralizing antibody negative patients. No significant association was found between the MXA genotype at these two SNPs and clinical, MRI and MXA gene expression in MS patients treated with IFN-beta therapy. PMID- 17126412 TI - Reproducibility of sonotubometry as Eustachian tube ventilatory function test in healthy children. AB - OBJECTIVES: To devise a simple and reliable diagnostic procedure to test Eustachian tube function routinely in an ENT outpatient setting. One method to measure ET ventilatory function is sonotubometry. The reproducibility of a recently updated sonotubometry set-up was tested in healthy children. METHODS: The test population comprised 61 school children aged from 6 to 8 years. Only otologically healthy children were included. Health state was established by means of a 12-item questionnaire. To test reproducibility, sonotubometric testing took place in two sessions of 10 acts of swallowing each. Spearman's coefficient was used to test the correlation between the two sets of measurements. All testing took place at a primary school in a nearby village. RESULTS: Opening of the ET was recorded in at least one of the two measurement sessions in 82% of the children. The first and second sessions were highly correlated, with a Spearman's coefficient of 0.89. CONCLUSIONS: In otologically healthy children, opening of the ET was recorded frequently using the updated sonotubometry set-up. Measurement results had high reproducibility. Therefore, the test forms a useful method to assess ET ventilatory function in otologically healthy children. The performance of this updated version needs to be established in children with otological diseases. PMID- 17126413 TI - High frequency distortion product otoacoustic emissions in children with and without middle ear dysfunction. AB - OBJECTIVE: Distortion product otoacoustic emissions (DPOAEs) (9-16kHz) are a useful measure of the function of the cochlea, which may be damaged by ototoxic drugs during anticancer chemotherapy. As children undergoing chemotherapy may also have middle ear problems, it is necessary to know if middle ear problems would have a confounding effect on the ability of DPOAEs to assess cochlear function in the extend high frequency region (9-16kHz). The present study aimed to investigate the effect of middle ear dysfunction on DPOAEs in the extended high frequency region in young children. METHODS: The sample was comprised of 100 ears of 50 school-aged children (21 boys and 29 girls) with a mean age of 6.3 years (S.D.=0.5; range 5.3-7.3). Otoscopy, pure tone hearing screening, tympanometry, acoustic reflexes and DPOAEs for both the conventional and extended high frequencies were administered to each child under typical school screening conditions. Participants were classified into one of three groups based on immittance (tympanometry and acoustic reflex) results. They included a "pass immittance" group, a "fail immittance" group and an "undetermined" group (with a pass in either tympanometry or acoustic reflexes, but not both). DPOAE amplitudes and signal-to-noise ratios (SNRs) were measured and compared across the three groups of participants. RESULTS: The fail immittance group showed significantly smaller DPOAE amplitudes and SNRs when compared to the other two groups at frequencies ranging from 1 to 9.5kHz and at 13kHz, but not at 10, 11, 12 and 14kHz. There was no significant difference in DPOAE results between the pass immittance and undetermined groups. CONCLUSIONS: Despite the adverse effects of middle ear dysfunction, its effect on DPOAEs in the extended high frequency region was not as severe as that in the lower frequency region. Hence, assessment of cochlear function in children with a middle ear lesion in the extended high frequencies using DPOAEs should be made with caution. PMID- 17126414 TI - A caution regarding the use of low-molecular weight heparin in pediatric otogenic lateral sinus thrombosis. AB - Lateral sinus thrombosis (LST), a rare complication of otitis media, is managed by antibiotics, surgery and anticoagulation. Traditionally, post-operative anticoagulation has been achieved by intravenous unfractionated heparin followed by oral warfarin. Fractionated, or low-molecular weight heparin derivatives (LMWH) have been introduced recently. There has been minimal literature to date regarding their use for the management of LST. We present use of the LMWH enoxaparin (Lovenox) for otogenic LST in two children, both of whom experienced hemorrhagic complications. On this basis and in the context of a literature review, we urge caution when using LMWH for pediatric otogenic LST. PMID- 17126415 TI - Acute herpes simplex laryngotracheitis: Report of two pediatric cases and review of the literature. AB - Herpes simplex virus (HSV) is a rare cause of laryngotracheitis (LT) with only 22 previously reported and confirmed pediatric cases in the literature. It is often associated with immune deficiency states and presents with a severe acute upper airway obstruction commonly requiring intensive care management and artificial ventilation. We present two cases of atypical laryngotracheitis in which HSV was found to be the causative pathogen subsequent to laryngoscopy and microbiologic investigations. While the first case was a previously well 8-month-old girl, the second was a 22-month-old immunosuppressed boy. One-third of the total confirmed cases in the English literature required intensive care management eliciting the setting as a pointer to the diagnosis. Laryngoscopy is a readily available tool for rapid diagnosis and controlled securing of the airway. This step may avoid controversial and potential counter-productive use of systemic steroids in these cases. PMID- 17126416 TI - Laboratory evaluation of a quantitative real-time reverse transcription PCR assay for the detection and identification of the four subgroups of avian metapneumovirus. AB - Avian metapneumovirus (AMPV) is an important pathogen causing respiratory diseases and egg drops in several avian species. Four AMPV subgroups have been identified. The laboratory diagnosis of AMPV infections relies on serological methods, on labour-intensive virus isolation procedures, and on recently developed subgroup specific reverse transcription PCR (RT-PCR) protocols. In the present study, both the specificity and sensitivity of a commercial real-time reverse transcription PCR (RRT-PCR) for the detection and identification of the four AMPV subgroups were evaluated. Fifteen non-AMPV avian viruses belonging to 7 genera and 32 AMPV belonging to the 4 subgroups were tested. No non-AMPV virus was detected, whereas all AMPV viruses were identified in agreement with their previous molecular and antigenic subgroup assignment. The sensitivity and quantitating ability of the RRT-PCR assay were determined using serial dilutions of RNA derived either from AMPV virus stocks or from runoff transcripts. In all cases, linear dose/responses were observed. The detection limits of the different subgroups ranged from 500 to 5000 RNA copies and from 0.03 to 3.16TCID50/ml. The results were reproducible under laboratory conditions, thus showing that quantitative RRT-PCR is a new and powerful tool for the rapid and sensitive detection, identification and quantitation of AMPVs. PMID- 17126417 TI - Characterization of field bovine herpesvirus samples using random amplified polymorphic DNA (RAPD). AB - Bovine herpesvirus 5 (BoHV-5) and Bovine herpesvirus 1 (BoHV-1) are members of the family Herpesviridae, subfamily Alphaherpesvirinae. Although BoHV-1 and BoHV 5 are closely related viruses (their genomes share 85% similarity), they exhibit some important differences at the genetic level which may explain the differences in their pathogenicity and epidemiological characteristics. Bovine herpesvirus 5 is a neurovirulent virus that causes fatal meningoencephalitis in calves and has been described in Australia, Argentina, United States and Brazil. BoHV-1 causes infectious bovine rhinotracheitis (IBR) and is reported around the world. The aim of this study was to establish the use of random amplified polymorphic DNA (RAPD) followed by phylogenetic analysis to characterize field samples of bovine herpesviruses. This technique was used to study the relationship of Brazilian samples of bovine encephalitis virus isolated in semen and the central nervous system (CNS), including BoHV-1.1 and BoHV-1.2 reference subtype virus for comparison. The samples tested could be clearly separated into BoHV-1.1, BoHV-1.2 and BoHV-5 by RAPD using two different primers. The results showed that RAPD analysis can be used as a diagnostic as well as a molecular epidemiological tool for herpesviruses. PMID- 17126418 TI - Development of AAV serotype-specific ELISAs using novel monoclonal antibodies. AB - Adeno-associated viruses (AAV) have been developed and evaluated as recombinant vectors for gene therapy. More recently, due to the advantages they offer for gene transfer, several AAV serotypes have gained increasing interest. However, monoclonal antibodies for the characterization and quantitation of vectors derived from different serotypes are at present not available. Serotype-specific monoclonal antibodies (mAbs) against the capsids of the serotypes 1/6, 4 and 5 are described. These antibodies, designated as ADK1a and b, ADK4 or ADK5a and b, reacted specifically with the indicated serotype capsids in cell lysates. ADK 1a and b cross-reacted with its highly related AAV6 serotype, but not with the other serotypes tested. The new antibodies recognized exclusively assembled capsids and neither free nor denatured capsid proteins as shown by fractionation experiments. In immunofluorescence experiments, the mAbs stained only distinct intranuclear foci in cells expressing the capsid protein. The development of capture ELISAs for quantitation of AAV1 and 6, AAV4 or AAV5 capsids illustrates that these novel monoclonal antibodies provide valuable tools for characterization of vector stocks. PMID- 17126419 TI - 5-HT2- and D1-mechanisms of the basolateral nucleus of the amygdala enhance conditioned fear and impair unconditioned fear. AB - The inferior colliculus (IC) is involved in processing of auditory information, but also integrates acoustic information of aversive nature. In fact, chemical stimulation of the IC with semicarbazide (SMC) - an inhibitor of the GABA synthesizing enzyme glutamic acid decarboxylase - has been found to cause defensive behavior in an open-field test and functions as an unconditioned stimulus in the place conditioned aversion test (PCA). A question has arisen regarding whether the basolateral nucleus of the amygdala (BLA) is involved in the acquisition of the aversive information ascending from the IC and whether dopaminergic and serotoninergic mechanisms of the BLA regulate this process. Recent evidence has shown that inactivation of the BLA with muscimol inhibits the PCA and causes an increase in the aversiveness of the chemical stimulation of the IC. Based on this, we examined the effects of ketanserin and SCH-23390, antagonists of the 5HT(2) and D(1) receptors, respectively, on the conditioned and unconditioned fear elicited by IC stimulation with SMC. The results obtained confirm the crucial role of 5-HT(2)- and D(1)-mechanisms of the BLA on conditioned fear in that ketanserin and SCH-23390 injections into the BLA caused a reduction in the PCA. On the other hand, ketanserin and SCH-23390 injections into the BLA enhanced the aversiveness of the IC injections of SMC. These findings suggest that while 5-HT(2) and DA(1) mechanisms in the BLA appear to facilitate the conditioned fear they inhibit the unconditioned fear triggered by IC activation. PMID- 17126420 TI - Impairments in spatial memory retention of GFAP-apoE4 female mice. AB - The human apolipoprotein E isoforms, apoE2, apoE3, and apoE4, have differential effects on brain function. Compared to apoE3, apoE4 increases the risk of age related cognitive decline in humans and female mice expressing apoE in neurons. Here, we show impaired spatial memory retention in female mice expressing apoE4 in astrocytes compared to those expressing apoE3 in astrocytes or lacking apoE. Thus, apoE4 impairs cognition whether expressed in neurons or astrocytes. PMID- 17126421 TI - Age-related changes in the motricity of the inbred mice strains 129/sv and C57BL/6j. AB - The development of motor skills was studied at different stages in the life of the mouse, focusing on three key aspects of motor development: early rhythmic motor activities prior to the acquisition of quadruped locomotion, motor skills in young adults, and the effect of aging on motor skills. The age-related development pattern was analysed and compared in two strains of major importance for genomic studies (C57Bl6/j and 129/sv). Early rhythmic air-stepping activities by l-dopa injected mice showed similar overall development in both strains; differences were observed with greater beating frequency and less inter-limb coordination in 129/sv, suggesting that 129/sv had a different maturation process. Performance on the rotarod by young adult C57Bl6/j gradually improved between 1 and 3 months, but then declined with age; performance on the treadmill also declined with an age-related increase in fatigability. Overall performance by 129/sv mice was lower than C57Bl6/j, and the age-related pattern of change was different, with 129/sv having relatively stable performance over time. Inter strain differences and their possible causes, in particular the role of dopaminergic pathways, are discussed together with repercussions affecting mutant phenotyping procedures. PMID- 17126422 TI - Analysis of telomere length variation in Leishmania over time. PMID- 17126424 TI - Characterization of a 60S complex of the adenomatous polyposis coli tumor suppressor protein. AB - The tumor suppressor protein adenomatous polyposis coli (APC) is a multifunctional protein with a well characterized role in the Wnt signal transduction pathway and roles in cytoskeletal regulation and cell polarity. The soluble pool of APC protein in colon epithelial tumor cells exists in two distinct complexes fractionating at approximately 20S and approximately 60S in size. The 20S complex contains components of the beta-catenin destruction complex and probably functions in the Wnt pathway. In this study, we characterized the molecular nature of the 60S APC- containing complex by examining known potential binding partners of APC. 60S APC did not contain EB1 or diaphanous, proteins that have been reported to interact with APC and are implicated in microtubule plus end stabilization. Nor did the two other microtubule associated proteins, MAP4 or KAP3, which is thought to link APC to kinesin motor proteins, associate with the 60S complex. Minor fractions of alpha-tubulin, gamma-tubulin and IQGAP1, a Rac1 and CDC42 effector that interacts with APC, specifically associated with APC in the 60S fraction. We propose that 60S APC is a discrete high molecular weight complex with a novel function in cytoskeletal regulation in epithelial cells apart from its well established role in targeting catenin destruction or its proposed role in microtubule plus end stabilization. PMID- 17126426 TI - Resolution of persistent late postoperative chylothorax after coil occlusion of aortopulmonary collaterals. AB - We report on successful treatment of persistent late postoperative chylothorax with coil occlusion of significant aortopulmonary collaterals in a young infant with functionally univentricular heart after bidirectional cavopulmonary connection. PMID- 17126427 TI - Particulate debris collected during carotid stenting: are we missing something? AB - Particulate and histopathologic examination of atherosclerotic material collected during carotid artery stenting is presented, illustrating the limitations of current knowledge regarding the use of distal protection devices (DPD) during this novel vascular intervention. PMID- 17126425 TI - Roles of the Raf/MEK/ERK pathway in cell growth, malignant transformation and drug resistance. AB - Growth factors and mitogens use the Ras/Raf/MEK/ERK signaling cascade to transmit signals from their receptors to regulate gene expression and prevent apoptosis. Some components of these pathways are mutated or aberrantly expressed in human cancer (e.g., Ras, B-Raf). Mutations also occur at genes encoding upstream receptors (e.g., EGFR and Flt-3) and chimeric chromosomal translocations (e.g., BCR-ABL) which transmit their signals through these cascades. Even in the absence of obvious genetic mutations, this pathway has been reported to be activated in over 50% of acute myelogenous leukemia and acute lymphocytic leukemia and is also frequently activated in other cancer types (e.g., breast and prostate cancers). Importantly, this increased expression is associated with a poor prognosis. The Ras/Raf/MEK/ERK and Ras/PI3K/PTEN/Akt pathways interact with each other to regulate growth and in some cases tumorigenesis. For example, in some cells, PTEN mutation may contribute to suppression of the Raf/MEK/ERK cascade due to the ability of activated Akt to phosphorylate and inactivate different Rafs. Although both of these pathways are commonly thought to have anti-apoptotic and drug resistance effects on cells, they display different cell lineage specific effects. For example, Raf/MEK/ERK is usually associated with proliferation and drug resistance of hematopoietic cells, while activation of the Raf/MEK/ERK cascade is suppressed in some prostate cancer cell lines which have mutations at PTEN and express high levels of activated Akt. Furthermore the Ras/Raf/MEK/ERK and Ras/PI3K/PTEN/Akt pathways also interact with the p53 pathway. Some of these interactions can result in controlling the activity and subcellular localization of Bim, Bak, Bax, Puma and Noxa. Raf/MEK/ERK may promote cell cycle arrest in prostate cells and this may be regulated by p53 as restoration of wild-type p53 in p53 deficient prostate cancer cells results in their enhanced sensitivity to chemotherapeutic drugs and increased expression of Raf/MEK/ERK pathway. Thus in advanced prostate cancer, it may be advantageous to induce Raf/MEK/ERK expression to promote cell cycle arrest, while in hematopoietic cancers it may be beneficial to inhibit Raf/MEK/ERK induced proliferation and drug resistance. Thus the Raf/MEK/ERK pathway has different effects on growth, prevention of apoptosis, cell cycle arrest and induction of drug resistance in cells of various lineages which may be due to the presence of functional p53 and PTEN and the expression of lineage specific factors. PMID- 17126428 TI - Torsades de pointes with a severely prolonged QT interval induced by an initial low dose sotalol intake. AB - Many drugs, including sotalol, have been implicated in prolonging QT interval and triggering torsades de pointes, a potentially fatal ventricular arrhythmia, especially during chronic therapy or in case of acute high dose toxicity. We report here a case with a severely prolonged QT interval and torsades de pointes after an initial intake of low dose sotalol (80 mg), indicating a probable inherent individual oversensitivity to sotalol. PMID- 17126429 TI - Transcatheter occlusion of the isolated scimitar vein anomaly camouflaged under dual pulmonary venous drainage of the right lung by the Amplatzer Ductal Occluder. AB - A 3.5-year-old girl, who presented recurrent pneumonia without discernible cyanosis or desaturation due to isolated scimitar vein anomaly camouflaged by dual pulmonary venous drainage of the right lung, underwent anterograde transvenous occlusion for this abnormal vessel successfully by the Amplatzer Ductal Occluder at 4 years old. PMID- 17126430 TI - A pedigree-analysis approach to the descriptive epidemiology of autosomal recessive disorders. AB - We describe a pedigree-analysis approach to estimating descriptive epidemiological parameters for autosomal-recessive disorders when the ancestral source of the disorder is known. We show that the expected frequency of carriers in a cohort equals the gene contribution of the ancestral source to that cohort, which is equivalent to the direct (additive) genetic relationship of that ancestor to the cohort. Also, the expected incidence of affected foetuses ranges from (1/2)F* to F*, where F* is the mean partial inbreeding coefficient (due to the ancestor) of the cohort. We applied this approach to complex vertebral malformation (CVM) in Holstein-Friesians in Australia, for which the ancestral source is a USA-born bull, Carlin-M Ivanhoe Bell. The estimated frequency of carriers was 2.47% for the 1992-born and 4.44% for the 1997-born cohort of Holstein-Friesian cows in Australia. The estimated incidence of affected foetuses/calves was considerably less than one per thousand, ranging from 0.0024 to 0.0048% for the 1992-born cohort, and from 0.0288 to 0.0576% for the 1997-born cohort. These incidences correspond to expected numbers of affected female foetuses/calves ranging from 2 to 4 for the 1992-born cohort and from 28 to 56 for the 1997-born cohort. This approach is easy to implement using software that is readily available. PMID- 17126431 TI - Tsetse control in cattle from pyrethroid footbaths. AB - In Burkina Faso, we assessed the efficacy of treating cattle with a footbath containing aqueous formulations of pyrethroids to control two tsetse-fly species, Glossina tachinoides Westwood, 1850 (Diptera, Glossinidae) and Glossina palpalis gambiensis Vanderplank 1949. Legs were the most targeted parts of the body for tsetse-fly blood meals: 81% (95% CI: 73, 89) for G. tachinoides and 88% (81, 95) for G. palpalis. The in-stable efficacy of footbath treatments was compared with manual full spraying with a 0.005% alphacypermethrin (Dominex, FMC, Philadelphia, USA) formulation (250mL versus 2L). The proportions of knocked-down flies were the same with footbath and full spray but the latter was more protective against fly bites. In field use, the efficacy of both methods should be similar given the recommended treatment frequency: 3 days for footbath versus 7 days for full spray. Among 96 cattle drinking at the same water point in Dafinso (Burkina Faso), 68 (71%) were treated with a footbath containing a 0.005% deltamethrin formulation (Vectocid, CEVA SA, Libourne, France). We observed the effect of this live-bait technique on the one hand on released cohorts of reared, irradiated flies, and on the other hand on wild tsetse flies. In both cases, the footbath treatment was associated with a reduction of the apparent fly density probably related to an increased mortality. PMID- 17126432 TI - Profiling of bacterial adhesin--host receptor recognition by soluble immunoglobulin superfamily domains. AB - Several gram-negative human pathogens recognize members of the carcinoembryonic antigen-related cell adhesion molecule (CEACAM) family. Pathogenic Neisseriae employ distinct isoforms of the colony opacity-associated proteins (Opa(CEA) proteins) to bind to the amino-terminal domains of CEACAMs. Here we present a novel approach to rapidly determine the CEACAM-binding properties of single bacteria. Expression of the isolated amino-terminal domains of various CEACAMs in eukaryotic cells yields soluble probes that selectively recognize Opa(CEA) expressing bacteria in a pull-down assay format. Furthermore, by expressing soluble CEACAMs as fusions to green-fluorescent protein (CEACAM-N-GFP), CEACAM binding bacteria can be decorated with a fluorescent label and analysed by flow cytometry allowing the specific detection of receptor binding events on the level of single bacteria. Besides its potential for rapid and quantitative analysis of pathogen-receptor interactions, this novel approach allows the detection of receptor recognition in heterogeneous bacterial populations and might represent a valuable tool for profiling the host binding capabilities of various microorganisms. PMID- 17126433 TI - A core curriculum for RTTs (radiation therapists/radiotherapy radiographers) designed for developing countries under the auspices of the international atomic energy agency (IAEA). PMID- 17126434 TI - Effect of age and radiation dose on local control after breast conserving treatment: EORTC trial 22881-10882. AB - PURPOSE: To determine whether the effect of an additional "boost" radiation after breast conservative therapy (BCT) on local control depends on age and evaluate the impact of a treatment policy with a threshold for age. PATIENTS AND METHODS: We used data from EORTC 22881-10882 trial, with median follow-up of 77.4 months. Patients receiving BCT and 50Gy whole breast irradiation were randomized to no boost and 16Gy boost (N=5318). RESULTS: In univariate analysis, a boost reduced local failure by a factor of 2 (P<0.0001). Multivariate analysis showed local control increased with age (P=0.0003). There was no evidence that the relative effect of a boost on local control depends on age (P=0.97) However in younger patients the 5-year local failure was higher, therefore the absolute reduction was greater. If the threshold-age for boost treatment were set at 40 years, 8.4% of the study population would receive a boost, resulting in a 5-year local failure of 6.1% in the study population. Changing the threshold-age to 60 years, 67% of the study population would receive a boost and the 5-year local failure would be reduced to 4.4%. CONCLUSIONS: In younger patients a boost dose resulted in a greater absolute reduction of local failure. The relative risk reduction was however similar for all ages. Applying a treatment policy with a threshold-age of 60 would result in 0.6% increase in local failure in the total study population, while sparing the boost to 1/3 of the patients. PMID- 17126435 TI - Differential involvement of regions of rostral prefrontal cortex (Brodmann area 10) in time- and event-based prospective memory. AB - Rostral prefrontal cortex (approximating Brodmann area 10) has been shown repeatedly to have a role in the maintenance and realization of delayed intentions that are triggered by event cues (i.e., event-based prospective memory). The cerebral organization of the processes associated with the use of time cues (time-based prospective memory) has however received less attention. In two positron emission tomography (PET) studies we therefore examined brain activity associated with time- and event-based prospective memory tasks. In the time-based condition of the first study, young healthy volunteers were asked to make a prospective response based on their self-estimation of the passage of time while engaged in an attention-demanding ongoing activity. In the time-based condition of the second study, participants had a clock available in the ongoing task display and did not need to estimate the time for the prospective response. In the event-based condition of both studies, participants were asked to make a prospective response when prospective cues were presented in ongoing trials. Both studies showed activation differences in rostral prefrontal cortex according to whether the task was time- or event-based. In study one, an area of left superior frontal gyrus was more active in the time-based condition. In study two, three rostral prefrontal regions were more active in the time-based condition: right superior frontal gyrus, anterior medial frontal lobe and anterior cingulate gyrus. A region in left superior frontal gyrus, different from the area found in the first study, was more active in the event-based condition. These results indicate involvement of multiple brain regions of rostral prefrontal cortex in time- and event-based prospective memory. The results are interpreted as reflecting the differing processing demands made by event- or time-based prospective memory tasks, and the differing demands of time-based tasks according to whether a clock is present as an aid. PMID- 17126436 TI - Brain regions and their dynamics in prospective memory retrieval: a MEG study. AB - We measured brain activity using magnetoencephalography in five participants during ongoing tasks that included prospective memory, retrospective memory, and oddball trials. Sources were identified in the hippocampal formation and posterior parietal and frontal lobes. Posterior parietal cortex activation had an earlier onset in the prospective memory condition than retrospective memory or oddball conditions, a higher level of theta activity in the retrospective condition, and higher levels of upper alpha in the prospective and oddball conditions. Activation of the hippocampal formation had a longer duration in the retrospective memory and prospective memory conditions than the oddball condition, but prominent alpha and theta band activity was present in all three conditions. We interpret the early (87 ms) onset of activity in parietal cortex as evidence for an initial noticing of appropriate conditions for a PM response. Hippocampal activity may reflect a subsequent memory search for the intended action. PMID- 17126437 TI - Vertical ocular responses to constant linear acceleration generated by fore-aft head translation in monkeys. AB - We examined the vertical linear vestibuloocular reflexes (LVORs) elicited by constant linear acceleration (0-0.5 g for >95 ms) during transient fore-aft translation in three monkeys. In the dark condition, small but consistent downward ocular responses to forward translation were observed (latencies >41 ms) when the initial vertical eye positions were at 0 degrees , although eye movements following backward translation were inconsistent among animals. These downward ocular responses showed the following three characteristics: they were independent of vertical gaze eccentricities, their magnitudes were almost proportional to the forward acceleration, and they were reduced by the large field (not the spot) visual information. These characteristics revealed that the downward ocular responses to forward translation were the tilt LVORs. In addition, we recognized that the translational LVOR, which depended on vertical gaze eccentricities, was working at the same time. Our data suggest that constant linear acceleration during forward translation evokes the tilt LVOR, as well as the illusory tilting perception. PMID- 17126438 TI - In vitro effect of quinolinic acid on energy metabolism in brain of young rats. AB - Quinolinic acid (QA) is found at increased concentrations in brain of patients affected by various common neurodegenerative disorders, including Huntington's and Alzheimer's diseases. Considering that the neuropathology of these disorders has been recently attributed at least in part to energy deficit, in the present study we investigated the in vitro effect of QA (0.1-100 microM) on various parameters of energy metabolism, such as glucose uptake, (14)CO(2) production and lactate production, as well as on the activities of the respiratory chain complexes I-V, the citric acid cycle (CAC) enzymes, creatine kinase (CK), lactate dehydrogenase (LDH) and Na(+),K(+)-ATPase and finally the rate of oxygen consumption in brain of 30-day-old rats. We initially observed that QA significantly increased glucose uptake (55%), whereas (14)CO(2) generation from glucose, acetate and citrate was inhibited (up to 60%). Furthermore, QA-induced increase of brain glucose uptake was prevented by the NMDA receptor antagonist MK 801. Complex II activity was also inhibited (up to 35%) by QA, whereas the other activities of the respiratory chain complexes, CAC enzymes, CK and Na(+),K(+) ATPase were not affected by the acid. Furthermore, inhibition of complex II activity was fully prevented by pre-incubating cortical homogenates with catalase plus superoxide dismutase, indicating that this effect was probably mediated by reactive oxygen species. In addition, lactate production was also not altered by QA, in contrast to the conversion of pyruvate to lactate catalyzed by LDH, which was significantly decreased (17%) by this neurotoxin. We also observed that QA did not change state III, state IV and the respiratory control ratio in the presence of glutamate/malate or succinate, suggesting that its effect on cellular respiration was rather weak. The data provide evidence that QA provokes a mild impairment of brain energy metabolism in vitro and does not support the view that the brain energy deficiency associated to certain neurodegenerative disorders could be solely endorsed to QA accumulation. PMID- 17126439 TI - Acidification decouples gap junctions but enlarges the receptive field size of horizontal cells in carp retina. AB - The receptive field size of retinal horizontal cells is much larger than their dendritic field size due to gap junctional coupling between the same sub-types of cell. Thus, horizontal cells form syncytia by electrical coupling. The basic receptive field profile of horizontal cells can be described by an exponential function based on measurement of responses to a slit of light moved tangentially from a recording electrode. The space constant of this exponential function is proportional to (g(s)/g(m))(1/2), where g(s) and g(m) represent gap junctional conductance and non-gap junctional conductance, respectively. Acidifying the superfusing solution by lowering the pH from 7.60 to 7.30 decreased the dye coupling, hyperpolarised the resting membrane potential and reduced the photoresponses of H1 type horizontal cells. Surprisingly, however, the receptive field size expanded significantly. Raising the pH from 7.30 to 7.60 or 7.90 produced opposite effects. These results were consistent with alkaline extracellular pH producing a greater increase in g(m) than in g(s) and enhancing release of transmitter from cones acting upon horizontal cells. PMID- 17126440 TI - A novel method for assessing the role of air in the microbiological contamination of poultry carcasses. AB - This paper describes a novel method of measuring the contamination of raw foods with airborne bacteria during primary processing. To demonstrate the approach, this study aimed to quantify the role of airborne bacteria in the contamination of broiler chicken carcasses undergoing processing in an evisceration room. Settle plates and broiler carcasses were exposed to the evisceration room air or to ultra-clean air provided by a high efficiency particulate air (HEPA) unit located within the room. The use of ultra-clean air reduced the total aerobic counts on horizontal settle plates by 68-fold, and on vertical settle plates by 14-fold. The use of ultra-clean air had no significant effect on the total aerobic counts on carcasses as measured by sponging (3.5 log(10) CFU cm(-2)) or skin excision (4.0 log(10) CFU cm(-2)). The novel approach was able to show that the carcasses entering the room were so heavily contaminated that the airborne bacteria in the evisceration room contributed less than 1% of the total numbers of bacteria on the carcasses. PMID- 17126441 TI - Antimicrobial susceptibility of enterococci strains isolated from slaughter animals on the data of Hungarian resistance monitoring system from 2001 to 2004. AB - Isolates of Enterococcus spp. were collected from January 2001 to December 2004 from caecal samples of slaughtered poultry, swine and cattle in Hungary. The isolates were identified by their growth and biochemical properties and with PCR. The antibiotic susceptibility of a total number of 1272 isolates was tested with disk diffusion test to ampicillin, gentamicin, streptomycin, tetracycline, erythromycin and vancomycin. It was established that although ampicillin and amoxicillin are often used in veterinary practice its resistance rate was relatively low. In the case of tetracyclines and macrolides, a high incidence of resistance was found. Susceptibility of strains to tetracyclines and/or macrolides reduced in both 2003 and 2004 in all animal species, which may be due to the more frequent usage of these drugs in the veterinary practice following the ban of growth promoters. The annual data of vancomycin resistance point to an association between the recovery of vancomycin-resistant enterococci (VRE) isolates and the use of avoparcin. This study indicates that reducing antimicrobial resistance in food animals could be possible with lower usage of antibiotics, although variations can occur with different strains. PMID- 17126442 TI - Efflux pump activity in fluoroquinolone and tetracycline resistant Salmonella and E. coli implicated in reduced susceptibility to household antimicrobial cleaning agents. AB - It has been shown that the inappropriate use of antimicrobial household agents selects for organisms with resistance mechanisms (e.g. efflux pumps), which could lead to the development of antibiotic resistance. The reverse hypothesis, that antibiotic-resistant organisms become tolerant to other antibacterial agents (e.g. disinfectants) due to the action of efflux pumps, has however not been extensively examined. The objective of this study was to establish whether there is a link between antibiotic resistance in potential gastrointestinal pathogens and reduced sensitivity of these organisms to commonly used household antimicrobial agents. In this study, tetracycline and ofloxacin sensitive and resistant Escherichia coli (9 strains) and Salmonella spp. (8 strains) were isolated from poultry and clinical samples. In order to assess whether these bacteria had active efflux pumps, ethidium bromide accumulation assays were performed. Extrusion of the active components of three commercial household agents (triclosan, sodium salicylate, and ortho-phenylphenol) by efflux pumps was tested using spectrophotometric accumulation assays. In order to simulate the kitchen environment, in-use disinfectant testing using the commercial household agents was performed to determine changes in their efficacy due to antibiotic resistance. Active efflux pump activity and extrusion of all three active ingredients was observed only in the antibiotic resistant organisms. The antibiotic sensitive bacteria were also more susceptible than the resistant isolates to the household antimicrobial agents at concentrations below that recommended by the manufacturer. These resistant bacteria could potentially be selected for and result in hard to treat infections. PMID- 17126443 TI - Genus specific features of bovine papillomavirus E6, E7, E5 and E8 proteins. AB - Six bovine papillomavirus (BPV) types, BPV-1 to -6, have been classified in genera Delta-papillomavirus (BPV-1 and -2), Epsilon-papillomavirus (BPV-5) and Xi papillomavirus (BPV-3, -4 and -6). In addition, 16 unclassified putative BPV types have been reported. In the present study, we characterized genus specific features of E6, E7, E5 (formerly E8) and E8 proteins of seven putative BPV types, BAPV-1, -2, -3, -4 and -10, BAA-5 and BPV-3c. These putative BPV types were classified in genera Epsilon- or Xi-papillomavirus. The E6 proteins of BPV and putative BPV types in Epsilon-papillomavirus showed high sequence similarities, and contained two typical zinc-binding domains. However, E7 proteins contained atypical zinc-binding domains, and lacked the canonical retinoblastoma tumor suppressor protein (pRB)-binding motif. BPV and putative BPV types in Xi papillomavirus contained E5 or E8 open reading frame (ORF) in the E6 position. The E5 ORFs encoded proteins consist of 42-amino acid with a hydrophobic transmembrane and a hydrophilic C-terminal domain. But the E8 ORFs encoded protein which have two transmembrane domains. Our results demonstrated that E5, E8, E6, E7 proteins of the putative BPV types, which are presumably classified in genera Epsilon- or Xi-papillomavirus, retained the some genus specific features. PMID- 17126444 TI - A strategy for efficient cross-presentation of CTL-epitope peptides leading to enhanced induction of in vivo tumor immunity. AB - The activation of antitumor cytotoxic T-lymphocytes (CTLs) depends on how efficiently the relevant tumor antigen peptides are delivered into the major histocompatibility complex (MHC) class I presentation pathway in antigen presenting cells (APCs). An elegant approach to promote the peptide-MHC class I association has been described for enhanced peptide transportation into the endoplasmic reticulum (ER) by adding an ER insertion signal sequence (Eriss). Nevertheless, this approach does not appear potent enough to induce in vivo tumor protective immunity. Herein, we present a novel peptide-vaccine strategy based on the combined utilization of Eriss and fusogenic liposomes (FLs) capable of directly introducing encapsulated CTL-epitope peptides into the MHC class I pathway of APCs. APCs pulsed with free peptides, FL-encapsulated peptides, or FL encapsulated Eriss-conjugated peptides exhibited comparable levels of antigen presenting activity at early phases after pulsing. Interestingly, whereas in the first two methods the APC ability began to decline 40 to 60 h after pulsing, FL encapsulated Eriss(+) peptides allowed APCs to retain peptide-presentation activity for at least 140 h. This advantage of FL-encapsulated Eriss(+) peptides correlated with the induction of more potent antitumor immunity compared with soluble Eriss(+) or Eriss(-) peptides or FL-encapsulated Eriss(-) peptides when they were administered in vivo. Thus, Eriss-conjugated CTL-epitope peptides encapsulated in FLs provide a highly efficient tumor-vaccine to enhance the induction of in vivo tumor immunity. PMID- 17126445 TI - Lectin binding studies on C-28/I2 and T/C-28a2 chondrocytes provide a basis for new tissue engineering and drug delivery perspectives in cartilage research. AB - The present study was performed to evaluate the applicability of plant lectins as mediators of bioadhesion in cartilage research using human chondrocyte cell lines C-28/I2 and T/C-28a2. The bioadhesive properties of fluorescein-labelled lectins with different carbohydrate specificities were investigated by flow cytometry. Specificity of the lectin-cell interactions was ascertained by competitive inhibition using complementary carbohydrates. As compared to that of other lectins, the interaction between wheat germ agglutinin (WGA) and chondrocytic cells was characterised by remarkable cytoadhesion, adequate binding strength and a high degree of specificity for N-acetyl-glucosamine as contained in hyaluronan chains. We therefore suggest WGA to be a promising candidate for mediating bioadhesion to low-adhesive scaffolds in cartilage tissue engineering. Moreover, the WGA-association rate of C-28/I2 and T/C-28a2 cells was dependent on temperature indicating cellular uptake of membrane-bound WGA. Intracellular enrichment was confirmed by confocal microscopy. Equilibration of intracellular pH gradients with monensin resulted in the reversal of quenching effects indicating accumulation of WGA within acid compartments of chondrocytic cells. Thus, WGA might be internalised into chondrocytes together with hyaluronan via the CD44 receptor-mediated endocytosis pathway and accumulated within lysosomes. This physiological process could represent a feasible pathway to target WGA functionalised drug delivery devices into chondrocytes. PMID- 17126446 TI - Complete regression of xenografted human carcinomas by a paclitaxel-carboxymethyl dextran conjugate (AZ10992). AB - Clinically available taxanes, such as paclitaxel and docetaxel, represent one of the most promising classes of anticancer agents, despite their toxicity. To improve their pharmacological profiles, AZ10992 was synthesized based on the concept that a rational design of a polymer-drug conjugate would increase the efficacy of the parent drug. This prodrug is a paclitaxel-carboxymethyl dextran conjugate (molecular weight 150,000 g/mol) via a gly-gly-phe-gly linker. The in vivo antitumor study using AZ10992 against colon26 carcinoma cells, resistant to paclitaxel, supported this concept. Additionally, the comparative efficacy studies of AZ10992 and paclitaxel using a panel of human tumor xenografts in nude mice showed the advantages of drug-polymer conjugation. The maximum tolerated dose of AZ10992 was more than twice as high as the MTD of paclitaxel. A repeated intravenous administration of AZ10992 at 30 mg/kg/day (five injections for 4 days) showed complete regression of MX-1 mammary carcinoma xenografts. Also, HT 29 colorectal tumor xenografts, which are highly refractory to paclitaxel, showed complete regression after AZ10992 administered at 30 mg/kg/day (seven injections for 4-days). Pharmacokinetic studies showed that there were significant increases in the amount and the exposure time of total paclitaxel in the tumors after intravenous administration of AZ10992, which explains the enhanced efficacy of AZ10992. PMID- 17126447 TI - Treatment of diabetic ketoacidosis in adults in Denmark: a national survey. AB - The aims of this study were to investigate management routines of diabetic ketoacidosis (DKA) in adult patients in departments of internal medicine in Denmark and to relate current routines of treatment to available evidence. A questionnaire requesting information on management routines of DKA was sent to all departments of internal medicine in Denmark responsible of managing DKA. Fifty-nine departments (88%) returned the questionnaire and/or a copy of their management protocol. At 19 departments (32%), all patients with DKA were managed in an intensive care unit (ICU). Twenty-four different insulin regimens and 21 fluid protocols were identified. Routines of insulin therapy varied in terms of doses and routes of administration. Fifty-eight departments (97%) used isotonic saline for hydration. Potassium supplements were administered as a separate infusion of either isotonic potassium-sodium-chloride (83%) or isotonic potassium chloride (10%). Recommended volumes to be administered during the first 8h of treatment varied significantly (median 4800ml, range 3750-7700ml). Use of bicarbonate was endorsed by 80%. This study shows significant variations in management routines of DKA in Denmark. In many cases, the treatment routines employed are not supported by evidence from clinical trials. We recommend implementation of national and/or European guidelines for management of DKA in adult patients. PMID- 17126448 TI - Predictive factors of early and sustained responses to peginterferon plus ribavirin combination therapy in Japanese patients infected with hepatitis C virus genotype 1b: amino acid substitutions in the core region and low-density lipoprotein cholesterol levels. AB - BACKGROUND/AIMS: We showed previously that amino acid (aa) substitutions in the HCV core region (HCV-CR) are predictors of non-virological response (NVR) to peginterferon (PEG-IFN) plus ribavirin (RBV) therapy. Here, we determined the predictive factors of sustained virological response (SVR) and early virologic response (EVR) to this treatment. METHODS: We evaluated the response to 48-week PEG-IFN-RBV therapy in 114 Japanese adults infected with HCV genotype 1b and determined the predictors of EVR and SVR. RESULTS: EVR was achieved by 70% and SVR by 45% of patients. 64% of patients who achieved EVR also showed SVR, while none of non-EVR achieved SVR. Multivariate analysis identified low-density lipoprotein cholesterol (LDL-C) (>or=86 mg/dl), aa substitutions in HCV-CR (double-wild-type; arginine at aa 70/leucine at aa 91), gamma-glutamyl transpeptidase (GGT) (<109 IU/l), RBV dose (>or=11.0mg/kg), and leukocyte count (>or=4500/mm3) as significant determinants of EVR, and aa substitutions in HCV-CR (double-wild-type), LDL-C (>or=86 mg/dl), male gender, ICG R15 (<10%), GGT (<109 IU/l), and RBV dose (>or=11.0 mg/kg) as determinants of SVR. Prediction of response to therapy based on combination of these factors had high sensitivity, specificity, positive, and negative predictive values. CONCLUSIONS: Our study identified aa substitutions in the core region and serum LDL-C as predictors of response to PEG-IFN-RBV therapy in Japanese patients infected with HCV genotype 1b. PMID- 17126449 TI - Polymorphisms of interleukin-1beta in Japanese patients with hepatitis B virus infection. AB - BACKGROUND/AIMS: Hepatitis B virus (HBV) induces liver cirrhosis (LC) and hepatocellular carcinoma (HCC) mainly by causing chronic necro-inflammatory hepatic disease. Our aim was to investigate the relationships between the polymorphisms of the interleukin-1B (IL-1B) promoter region and the interleukin-1 receptor antagonist gene (IL-1RN) and disease progression in an HBV-infected Japanese population. METHODS: Genomic DNA was extracted from the peripheral blood of 237 HBV carriers. Polymorphisms in IL-1B and IL-1RN were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and PCR with confronting two-pair primers (PCR-CTPP) methods. These polymorphic sites include the promoter regions of IL-1B at positions -511 and -31, and IL-1RN variable tandem repeats. RESULTS: The IL-1B -31 and -511 loci were in complete linkage disequilibrium, and the frequency of the IL-1B -31 T carrier (IL-1B -31 T/T or T/C) was significantly higher in HBV carriers with LC compared to those without LC (LC; 86.1% vs non-LC; 72.1%, P=0.009). There was no difference in the genotype distribution of the IL-1RN polymorphism. CONCLUSIONS: This is the first report describing the association between IL-1B polymorphism and HBV-related hepatic fibrosis, and our data suggest that IL-1B polymorphisms may be related to disease progression of HBV-related hepatitis in Japan. PMID- 17126451 TI - Adolescent screen-viewing behaviour is associated with consumption of sugar sweetened beverages: the role of habit strength and perceived parental norms. AB - The association between adolescent screen-viewing behaviour (i.e., television viewing and computer use) and the consumption of sugar-sweetened beverages was studied in a Dutch sample of adolescents (N=383) using self-administered questionnaires. In particular, the previously understudied role of habit and perceived parental norms in the execution of these behaviours was investigated. Results showed that screen-viewing behaviour was associated with consumption of sugar-sweetened beverages (r=.32). Habit strength of both behaviours correlated with a large effect size (r=.50). The interaction between both behaviours was underlined by the finding that consumption of sugar-sweetened beverages was explained by perceived parental norms regarding screen-viewing behaviour (beta=.12; adjusted for the behaviour and perceived parental norm regarding sugar sweetened beverage consumption). Consequences of the identified role of habit and parental norms in the interplay between sedentary behaviour and consumption of sugar-sweetened beverages among adolescents are discussed. PMID- 17126452 TI - Purine and pyrimidine nucleosides preserve human astrocytoma cell adenylate energy charge under ischemic conditions. AB - The brain depends on both glycolysis and mitochondrial oxidative phosphorylation for maintenance of ATP pools. Astrocytes play an integral role in brain functions providing trophic supports and energy substrates for neurons. In this paper, we report that human astrocytoma cells (ADF) undergoing ischemic conditions may use both purine and pyrimidine nucleosides as energy source to slow down cellular damage. The cells are subjected to metabolic stress conditions by exclusion of glucose and incubation with oligomycin (an inhibitor of oxidative phosphorylation). This treatment brings about a depletion of the ATP pool, with a concomitant increase in the AMP levels, which results in a significant decrease of the adenylate energy charge. The presence of purine nucleosides in the culture medium preserves the adenylate energy charge, and improves cell viability. Besides purine nucleosides, also pyrimidine nucleosides, such as uridine and, to a lesser extent, cytidine, are able to preserve the ATP pool. The determination of lactate in the incubation medium indicates that nucleosides can preserve the ATP pool through anaerobic glycolysis, thus pointing to a relevant role of the phosphorolytic cleavage of the N-glycosidic bond of nucleosides which generates, without energy expense, the phosphorylated pentose, which through the pentose phosphate pathway and glycolysis can be converted to energetic intermediates also in the absence of oxygen. In fact, ADF cells possess both purine nucleoside phosphorylase and uridine phosphorylase activities. PMID- 17126453 TI - On being of good character: nurse education and the assessment of good character. AB - The Nursing and Midwifery Council sets good character as a condition not only of entry to the UK register of nurses but also for entry to and continuation on pre registration nursing programmes in the UK. This requires education providers to assess the character of potential and actual nursing students yet the guidance offered by the NMC for making such assessments is insufficiently developed and serves to provide criteria by which determination can be made of good character only in a narrow sense. Assessment of character requires making value judgements about others and lends support to the idea that nurse education has an explicit moral component. This paper outlines some of the difficulties in assessing character, explores the idea that good character can be assessed in both a thin and a thick sense, and considers some of the consequent implications for nurse educationalists. PMID- 17126454 TI - Orientation of nurses towards formal and informal learning: motives and perceptions. AB - The aim of this exploratory study was to gain information on the current orientation of registered nurses towards continuing education and lifelong learning. The population (N=162) consists of 2nd and 1st Level nurses who have or are currently taking part in continuing education. Qualitative empirical data were obtained through semi structured one to one interviews. The research questions sought information related to some of the reasons and motives for the participants' taking part in various categories of learning. Also explored was what factors might influence these participants' learning activities and the views and perceptions of their learning experiences. For many of these nurses, the initial motive for taking part in continuing education was the perception that they were being left behind by the higher educational level of nurses entering the profession. Contrary to some anecdotal views, the participants generally felt that higher education (HE) contributed to enhanced client care, reporting additional personal and professional satisfaction. Alleged poor support from managers for continuing education and the lack of parity, often within the same organisation regarding the selection criteria to take part in a variety of learning activities, was a source of dissatisfaction for some of these participants. Their determination to learn, however, remained strong. PMID- 17126450 TI - Polymer genomics: an insight into pharmacology and toxicology of nanomedicines. AB - Synthetic polymers and nanomaterials display selective phenotypic effects in cells and in the body signal transduction mechanisms involved in inflammation, differentiation, proliferation, and apoptosis. When physically mixed or covalently conjugated with cytotoxic agents, bacterial DNA or antigens, polymers can drastically alter specific genetically controlled responses to these agents. These effects, in part, result from cooperative interactions of polymers and nanomaterials with plasma cell membranes and trafficking of polymers and nanomaterials to intracellular organelles. Cells and whole organism responses to these materials can be phenotype or genotype dependent. In selected cases, polymer agents can bypass limitations to biological responses imposed by the genotype, for example, phenotypic correction of immune response by polyelectrolytes. Overall, these effects are relatively benign as they do not result in cytotoxicity or major toxicities in the body. Collectively, however, these studies support the need for assessing pharmacogenomic effects of polymer materials to maximize clinical outcomes and understand the pharmacological and toxicological effects of polymer formulations of biological agents, i.e. polymer genomics. PMID- 17126455 TI - Reasons for post registration learning: impact of the learning experience. AB - This exploratory study is the second stage of a multi-method approach to explore some of the reasons why registered nurses take part in formal and informal learning and higher education. Also under exploration is the extent to which their various learning experiences met their expectations. The sample consists of 25 registered nurses, randomly selected from a population (N=162) of 1st and 2nd Level nurses employed in public and private clinical settings, who have taken part in continuing education to obtain a Diploma or a Degree in Nursing. The data gathering was achieved by three focus group interviews. Formal learning included mandatory and non-mandatory in house teaching. Mandatory input was regarded as a self-protection measure by the employer, not related to personal development and with no perceived benefit on client care. Regarding the achievement of additional clinical competences, the overwhelming reason is to improve client care, although the motives of the employer and medical staff are viewed with some suspicion as a measure to saving doctors' time. Higher education pursuits appears to be triggered by a perceived need to increase the participants' level of existing knowledge in line with that of current students on placement, but adjustment to adult life transition period also appear to be an influencing factor. In terms of learning experiences, the relevance and quality of mandatory sessions came under criticism, whilst a measure of surprise was expressed at the unexpected benefits, at a personal and a professional level, gained by taking part in academic learning. PMID- 17126456 TI - Digital diagnostics: Three-dimensional modelling. AB - Three-dimensional imaging techniques, such as computed tomograms (CT), structured light, and stereophotogrammetry, can be used to capture three-dimensional coordinate data, but comprehensive analysis is required to transform these techniques into powerful diagnostic tools. The object of this review is to highlight analytical functionality using software developed to study three dimensional digital imaging and communications in medicine (DICOM) based digital data for diagnosis, planning of treatment, and evaluation of craniofacial changes. My specific aim was to apply three-dimensional software routines using geometric morphometrics or conventional measurements. These routines rely on robust algorithms to construct mean objects by manipulating the three-dimensional x, y, and z coordinates of all the objects' vertices. Conventional measurements and statistical tests can then be applied to the changes in the vertices, say, before and after treatment. Using graphical and geometric morphometric techniques such as finite-element analysis and principal components analysis, clinical craniofacial modelling can be used for the localisation and quantification of soft and hard tissue changes; diagnostic modelling can be undertaken for planning of treatment, and data-driven predictive modelling can be undertaken for the planning of many procedures based on the surgeon's own experience, patients, and resources. Three-dimensional modelling of digital data may therefore have added value for clinical diagnosis, and planning and assessment of treatment, including audit. PMID- 17126457 TI - A Q methodology study of women's experience of enduring postnatal perineal and pelvic floor morbidity. AB - OBJECTIVE: to explore women's experiences of enduring postnatal perineal and pelvic floor morbidity. DESIGN: a retrospective Q methodology study of postnatal women between 12 and 18 months postpartum. SETTING: maternity services at the Heart of England NHS Foundation Trust, Birmingham. PARTICIPANTS: women were self identified by expression of interest as part of their earlier involvement in a retrospective cross-sectional community survey of enduring postnatal perineal and pelvic floor morbidity. Twenty women responded to the invitation posted on the back of the initial survey questionnaire. All 20 women took part in stage one (interview) of the study (100% response rate). Fourteen women returned the completed response grids in stage four after 1 month of dissemination (response rate 70%). METHODS: the project included five stages: (1) data were gathered using an individual face-to-face, semi-structured, community-based interview method (developing the concourse); (2) the concourse was analysed using MAXqda (2004) to produce 'themes'; (3) the themes were reduced to 'statements' that reflected the overall content of the concourse using an unstructured evolving approach to Q set design (giving the Q set); (4) participants were asked to sort the statements (Q sorting) according to a pre-designed distribution grid (providing individual participant response grids); and (5) the response grids were factor analysed using PQMethod (V2.0), which generates clusters of participants rather than clusters of variables. Factor loadings were calculated using factor analysis by principal components with varimax rotation. This produced a list of factors, each of which represents a 'story' of women's experience of enduring postnatal perineal and pelvic floor morbidity. FINDINGS: five factors were identified: perineal morbidity of minor inconvenience, insufficient support and services, the 'taboo' subject of enduring perineal and pelvic floor morbidity, normalising morbidity and the isolation of perineal morbidity. Women varied in their experiences of enduring postnatal perineal and pelvic floor morbidity, with some having minor problems, with little affect on daily living, whereas others were significantly affected. The affect of the morbidity varied. Women highlighted the lack of service provision for these problems and stated that health-care practitioners, and society at large, were often dismissive of, or trivialised, their experiences of enduring postnatal perineal and pelvic floor morbidity. CONCLUSION: the findings from this study highlight that the postnatal recovery period is longer than the presumed 6 weeks. The effect of postnatal perineal and pelvic floor morbidity on women's lives varies, and more long-term postnatal health support and research using women centred outcomes is needed. PMID- 17126458 TI - A descriptive account of New Zealand mothers' responses to open-ended questions on their breast feeding experiences. AB - OBJECTIVE: To describe the breast feeding experiences of mothers in New Zealand. DESIGN: Descriptive, qualitative study. After a previous quantitative analysis of a questionnaire, the open-ended responses by the women were examined using a thematic analysis approach. SETTING: Hawke's Bay, New Zealand. PARTICIPANTS: 153 primiparous and multiparous breast feeding women aged between 20 and 49 years, who had had a caesarean section or vaginal birth between 4 months and 3 years previously. FINDINGS: Themes identified during the data analysis are as follows: persistence, determination, confidence and satisfaction; pain and limitation of mothering activities; conflicting advice and professionalism; and others' expectations. CONCLUSIONS: Breast feeding experience could be detrimentally affected by physical factors, inconsistent health-professional support and others' expectations. Negative influences, however, were likely to be overcome by persistence, determination, confidence and satisfaction. IMPLICATIONS FOR PRACTICE: Implications for midwifery practice from the study iterate the importance of consistency of advice and skills of health professionals, of listening and understanding women's responses to early breast feeding assistance and giving continuous encouragement and support throughout their breast feeding experience. To meet these recommendations, breast feeding education for health professionals needs to include technical expertise and communication skills. Recommendations for further research from this study include investigations into why women commit to breast feeding and an exploration of women's perceptions of midwives' breast feeding support. PMID- 17126459 TI - A description of Swedish midwives' reflections on their experience of caring for teenage girls during pregnancy and childbirth. AB - OBJECTIVE: to describe Swedish midwives' reflections on their experiences of caring for teenage girls during pregnancy and childbirth. DESIGN: a hermeneutical approach was used, with focus-group discussions as the method of data collection. SETTING: three focus-group discussions were conducted in a county comprising urban, suburban and rural districts in south-western Sweden. PARTICIPANTS: a voluntary sample of 24 midwives, aged 32-61 years, who were caring for women in different birth settings, participated in 2004. FINDINGS: two main themes, with three sub-themes each, were generated by the midwives: (1) the teenage mother, as 'teenagers who are proud of becoming mothers', 'teenagers who are unprepared for becoming mothers' and 'teenagers with an immigrant background', and (2) 'the midwives' wish to care for the teenage mother', as 'taking the teenage mother seriously', 'being an important person for the teenage mother', and 'being a help for the teenage mother'. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: the findings provide some understanding of the unique characteristics of caring for teenage mothers. For the midwives, the most important aspects of caring for the teenage mother included taking the teenage mother seriously, allowing the midwife herself to become an important person for the teenage mother, and being a help to the mother. This help requires the midwife to balance the different needs of each teenage mother. PMID- 17126460 TI - Reconstruction of the digital flexor pulley system: a retrospective comparison of two methods of treatment. AB - The rare injury of closed rupture of the A2 and A3 flexor pulleys was treated using two non-encircling techniques of pulley reconstruction. Thirteen patients were treated with an extensor retinaculum graft (Group A). At a mean follow-up time of 48 months, the average PIP flexion was 97%, the power grip strength 96%, the pinch grip strength 100% and the thickening 94% of the uninjured side. Ten patients were treated with a free palmaris longus tendon grafts (Group B). At a mean follow-up time of 57 months, the average PIP flexion was 94%, the power grip strength 98%, the pinch grip strength 100% and the thickening 95% of the uninjured side. In both groups, finger extension was unrestricted. The Buck Gramcko score included 10 excellent, two good and one fair result in Group A and seven excellent, two good and one fair result in Group B. Both techniques proved beneficial. All climbers returned to their previous standard and all non-climbers regained full finger dexterity in their previous job. PMID- 17126461 TI - Hip, knee, ankle kinematics and kinetics during stair ascent and descent in healthy young individuals. AB - BACKGROUND: Few studies have reported the biomechanical aspects of stair climbing for this ergonomically demanding task. The purpose of this ethically approved study was to identify normal functional parameters of the lower limb during stair climbing and to compare the actions of stair ascent and descent in young healthy individuals. METHODS: Thirty-three young healthy subjects, (16 M, 17 F, range 18 39 years) participated in the study. The laboratory staircase consisted of four steps (rise height 18 cm, tread length 28.5 cm). Kinematic data were recorded using 3D motion analysis system. Temporal gait cycle data and ground reaction forces were recorded using a force platform. Kinetic data were standardized to body mass and height. FINDINGS: Paired-samples t tests showed significantly greater hip and knee angles (mean difference standard deviation (SD): hip 28.10 degrees (SD 4.08), knee 3.39 degrees (SD 7.20)) and hip and knee moments (hip 0.25 Nm/kg (SD 0.18), knee 0.17 Nm/kg (SD 0.15)) during stair ascent compared to descent. Significantly greater ankle dorsiflexion angles (9.90 degrees (SD 3.80)) and plantarflexion angles (8.78 degrees (SD 4.80)) were found during stair descent compared to ascent. Coefficient of variation (mean (SD)) in percentage between repeated tests varied for joint angles and moments, respectively (2.35% (SD 1.83)-17.53% (SD 13.62)) and (4.65% (SD 2.99)-40.73% (SD 24.77)). INTERPRETATION: Stair ascent was shown to be the more demanding biomechanical task when compared to stair descent for healthy young subjects. The findings from the current study provide baseline measures for pathological studies, theoretical joint modelling, and for mechanical joint simulators. PMID- 17126462 TI - Re-expressing the division of British medicine under the NHS: the importance of locality in general practitioners' oral histories. AB - This paper, based on original oral history research in a single locality, re examines the impact of the structural division of British medicine, especially between community and hospital-based medicine, on rank-and-file general practice. Interviews were carried out with 29 retired and practising National Health Service (NHS) general practitioners (GPs) in Paisley, Scotland. In contrast to the historiography and literature of academic general practice, most retired and working family doctors who were interviewed rejected the significance of the division, and instead placed emphasis on the positive relationships between primary and secondary care in their locality. Particular stress was placed by these interviewees on the significance of local medical relationships in crossing the divide between primary and secondary medical practice. An analysis of the oral testimonies, informed by the work of Bakhtin [(1984). Problems of Dostoevsky's poetics. Minnesota: University of Minnesota Press] and Schrager [(1983). What is social in oral history? International Journal of Oral History, 4(2), 76-98] identified multiple discourses within single accounts. These discourses contained 'dialects' that suggested a commonality of beliefs, especially when social and clinical networks were detailed, but they also contained 'traces' of oppositional narratives. These 'traces' of alternative perspectives suggest both the power of imagined solidarity and the exclusion of a minority of practitioners. Thus imagined solidarities, based on the making of local identities, combined with imagined oppositional positions, in which those who were constructed as not belonging were given voice. We argue that, to understand alliances between community and hospital medicine it is necessary to appreciate the position of the excluded. Those family doctors who were identified in the oral histories as outsiders tended to serve the more deprived populations. The role of private medicine was especially significant in the process of exclusion, especially the pattern of general practice referrals of patients from the NHS to secondary private care. PMID- 17126463 TI - Issues in exploring variation in childhood socioeconomic gradients by age: a response to Case, Paxson, and Vogl. PMID- 17126464 TI - Assisting or compromising intervention? The concept of 'culture' in biomedical and social research on HIV/AIDS. AB - This paper addresses how the notion of 'culture' has been understood and employed by both epidemiologists and anthropologists with respect to the literature on HIV/AIDS in Sub-Saharan Africa. It examines the shift towards non-biomedical understandings of the epidemic. The concept of 'culture' has been 'bandied about' [Schoepf, B.G. (2004). AIDS, history and struggles over meaning. In E. Kalipeni, S. Craddock, J.R. Oppong, & J. Ghosh (Eds.), HIV and AIDS in Africa: Beyond epidemiology. Oxford: Blackwell.], and yet no authors in the literature reviewed here attempt a more systematic account of the 'bandying about' itself. This paper thus attempts to address and close this gap. For biomedical researchers and epidemiologists, broadly speaking, 'culture' appears to compromise intervention, whilst for medical anthropologists, 'culture' is often seen as having the potential to assist intervention. 'Culture' comes to be multifaceted and laden with varying assumptions, which range from 'culture' being bounded and timeless, to 'culture' being linked to macro-processes, historically shaped, and contested. In turn, 'culture' has variously been understood as both the cause of, and solution to, the epidemic. It is also understood as having structured local interpretations of, and responses to, the epidemic. At the same time as noting the apparent dichotomy between the biomedical and social approaches, however, the models may inadvertently share certain assumptions about 'culture' as an essentializing signifier of difference. To this extent HIV/AIDS research could be improved by incorporating wider anthropological debates about the problems of the 'culture' concept. PMID- 17126465 TI - Interpreters as co-diagnosticians: overlapping roles and services between providers and interpreters. AB - This study examined medical interpreters' practice of the co-diagnostician role and further explored its practical, institutional, and ethical implications. Twenty-six professional interpreters (of 17 languages), 4 patients, and 12 health care providers were recruited for this study, which involves participant observation and interviews undertaken in the Midwestern US. Constant comparative analysis was used to develop themes of interpreters' communicative practices. Interpreters justified their role performances by claiming the identity of a member of the health care team and their work as part of the team effort. Their communicative strategies as a co-diagnostician reflect their preconception of the social hierarchy of health-care settings and the emphasis on diagnostic efficacy. I have identified five strategies for the co-diagnostician role. These were assuming the provider's communicative goals; editorializing information for medical emphasis; initiating information-seeking behaviors; participating in diagnostic tasks; and volunteering medical information to the patients. Although many strategies can be attributed to interpreters' effort to conserve providers' time and to bridge the cultural differences, they also pose risks to patients' privacy, clinical consequences, and provider-patient relationships. PMID- 17126466 TI - Prevalence of mental disorders in children and adolescents from a Spanish slum. AB - This paper reports rates of psychopathology in a population of 9- and 13-yr olds from a Spanish slum. Two cohorts of all the children born in 1989 and in 1993 and registered in the census of a municipality in 2001 were assessed over a 3-yr period with structured diagnostic interviews and functional measures. In the first year of the study 79 (53.7%) children of the adolescent 13-yr-old population and 72 (59.5%) of the pre-adolescent 9-yr-old population participated. Between 30% and 60% of preadolescents and between 30% and 50% of adolescents presented some mental disorder. Anxiety and disruptive behavior disorders were the most frequent disorders in both cohorts. For both genders, the highest risk for any psychopathology was at 10 yr. We found that, psychopathology and functional impairment decreased with age, and that the psychopathology of children in a peripheral slum of a big city is 3 times higher than the median of the general population. This information should be useful for administrators providing services for children from the most disadvantaged segment of the population. PMID- 17126467 TI - Comparative developmental toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin in the hamster, rat and guinea pig. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a persistent environmental contaminant capable of causing a wide variety of adverse health effects including teratogenesis and altered development. The objective of this study was to compare the developmental toxicity of TCDD in the hamster, rat and guinea pig, which in mature animals exhibit a relatively low, medium and high sensitivity to TCDD, respectively. A single oral dose of TCDD was administered to pregnant rats (0, 1.5, 3.0, 6.0 or 18.0microg/kg) on gestation day 10, pregnant hamsters (0, 1.5, 3.0, 6.0 or 18.0microg/kg) on gestation day 9 and pregnant guinea pigs (0, 0.15 or 1.5microg/kg) on gestation day 14 with fetal analysis on gestation day 20, 15 and 56, respectively. The developmental toxicity of TCDD in the three species included increased fetal mortality, alterations to fetal body weight, body length, organ weight and significant changes to the fetal white blood cell differential counts. Additionally, teratogenic responses were observed in the hamster and rat consisting of cleft palate, kidney congestion, hydronephrosis and intestinal hemorrhaging. Furthermore, the results from this study demonstrate that despite the up to 5000-fold interspecies variability to the acute lethal potency of TCDD observed in mature guinea pigs, rats and hamsters, the developing fetus is uniquely vulnerable to gestational TCDD exposure and displays approximately a 10-fold variability in fetal lethal potency in these species. Together, these results will assist efforts to reduce the uncertainty in the risk assessment for TCDD in sensitive populations, such as the developing embryo and fetus. PMID- 17126468 TI - Evaluation of cognitive and biochemical effects of low-level exposure to sarin in rhesus and African green monkeys. AB - We investigated the potential of low-level exposures to the chemical warfare nerve agent, sarin, to produce adverse effects. Rhesus (Macaca mulatta) and African green monkeys (Chlorocebus acthiops) were trained on a serial probe recognition (SPR) task before IM administration of a low-level concentration (5.87 microg/kg or 2.93 microg/kg) of sarin. Blood was sampled before agent administration and at various times following administration. Sarin administration did not disrupt performance on the SPR task in either species. Major dependent measures characterizing performance (accuracy, number of completed trials per session, average choice response time) were largely unaffected on the day sarin was administered as well as on subsequent testing sessions occurring over several weeks following administration. Analyses of red blood cell (RBC) and plasma samples revealed that sarin administration produced a substantial degree of inhibition of circulating acetylcholinesterase (AChE) in RBC fractions and butyrylcholinesterase (BChE) in plasma fractions, which only slowly recovered. In this regard, AChE activity was inhibited to a greater extent than BChE activity. Blood samples were also evaluated for regenerated sarin, which was found in RBC and plasma fractions in both species and showed orderly elimination functions. More sarin was regenerated from RBC fractions than from plasma fractions. Elimination of regenerated sarin was much slower in RBC than plasma and exceeded the expected time of AChE aging, suggesting the presence of additional sarin binding sites. In general, effects were similar in both species. Taken together, our results show that while the concentrations of sarin administered were clearly biochemically active, they were below those that are required to produce a disruption of behavioral performance. PMID- 17126469 TI - Effect of acetaminophen administration to rats chronically exposed to depleted uranium. AB - The extensive use of depleted uranium (DU) in both civilian and military applications results in the increase of the number of human beings exposed to this compound. We previously found that DU chronic exposure induces the expression of CYP enzymes involved in the metabolism of xenobiotics (drugs). In order to evaluate the consequences of these changes on the metabolism of a drug, rats chronically exposed to DU (40mg/l) were treated by acetaminophen (APAP, 400mg/kg) at the end of the 9-month contamination. Acetaminophen is considered as a safe drug within the therapeutic range but in the case of overdose or in sensitive animals, hepatotoxicity and nephrotoxicity could occur. In the present work, plasma concentration of APAP was higher in the DU group compared to the non contaminated group. In addition, administration of APAP to the DU-exposed rats increased plasma ALT (p<0.01) and AST (p<0.05) more rapidly than in the control group. Nevertheless, no histological alteration of the liver was observed but renal injury characterized by incomplete proximal tubular cell necrosis was higher for the DU-exposed rats. Moreover, in the kidney, CYP2E1 gene expression, an important CYP responsible for APAP bioactivation and toxicity, is increased (p<0.01) in the DU-exposed group compared to the control group. In the liver, CYP's activities were decreased between control and DU-exposed rats. These results could explain the worse elimination of APAP in the plasma and confirm our hypothesis of a modification of the drug metabolism following a DU chronic contamination. PMID- 17126470 TI - Analysis of neurosterols and neurosteroids by mass spectrometry. AB - In man the brain represents about 2% of the body weight, but contains 25% of the body's cholesterol. Cholesterol itself does not cross the blood-brain barrier and is synthesised in situ. Excess cholesterol from brain is exported in the form of oxysterols, or metabolised to steroids, which in contrast to cholesterol can cross the blood-brain barrier. Steroids and oxysterols may be synthesised in brain, but can also be transported into brain from peripheral tissue. Both oxysterols and steroids have biological activity in brain. They can behave as ligands for classical nuclear receptors, and exert their effects over hours to days, or interact with neurotransmitter gated ion channels and modulate neural transmission exerting their effects in milliseconds. The exact sterol and steroid content of brain has yet to be thoroughly characterised. In this mini-review we will discuss mass spectrometry methods for the analysis of steroids and sterols in brain, and propose methods suitable for the profiling of different brain regions with high sensitivity (sub pg) and specificity. PMID- 17126471 TI - Do we taste fat? AB - Sense of taste informs the body about the quality of ingested foods. Five sub modalities allowing the perception of sweet, salty, sour, bitter, and umami stimuli are classically depicted. However, the inborn attraction of mammals for fatty foods raises the possibility of an additional orosensory modality devoted to fat perception. For a long time, dietary lipids were thought to be detected only by trigeminal (texture perception), retronasal olfactory, and post-ingestive cues. This minireview analyses recent findings showing that gustation also plays a significant role in dietary lipid perception. PMID- 17126472 TI - Does the type of out-of-hospital airway interfere with other cardiopulmonary resuscitation tasks? AB - BACKGROUND: Out-of-hospital rescuers often perform tracheal intubation (TI) prior to other cardiopulmonary resuscitation (CPR) interventions. TI is a complex and error-prone procedure that may interfere with other key resuscitation tasks. We compared the effects of TI versus esophageal tracheal combitube (ETC) insertion on the accomplishment of other interventions during simulated cardiopulmonary resuscitation. METHODS: In this prospective trial using a human simulator, two paramedic teams simulated resuscitation of a ventricular fibrillation cardiopulmonary arrest using standard Advanced Cardiac Life Support guidelines. In each of two trials, teams used either TI or ETC as the primary airway device. Following delivery of three rescue shocks, we measured time intervals to successful airway placement, intravenous (IV) line insertion, drug administration, delivery of fourth rescue shock and completion of all four tasks. We also measured the total time without chest compressions. We compared task completion times using non-parametric statistics (Wilcoxon signed-ranks test) with a Bonferroni-adjusted p-value of 0.008. RESULTS: Twenty teams each completed two scenarios. Participants required a median of 172.5 s (IQR: 146.5-225.5) to accomplish all four tasks. Elapsed time to airway placement was significantly less for ETC than TI (median difference 26.5 s (IQR 13-44.5), p=0.002). Time without chest compressions was less for ETC than TI (median difference 8.5 s (IQR 2.5-23.5), p=0.005). There were no differences between ETC and TI in times to IV placement (median difference 23.5 s (IQR -20 to 61), p=0.11), drug delivery (39.5 s (IQR -18 to 63), p=0.07), delivery of fourth rescue shock (39.5 s (IQR -21.5 to 87.5), p=0.07) or completion of all four tasks (33 s (IQR -11 to 74.5), p=0.08). CONCLUSION: Compared with TI, ETC reduced time to airway placement and time without chest compressions, but did not affect elapsed times to accomplish other interventions. Additional time differences may be realized if translated to clinical out-of-hospital conditions. PMID- 17126473 TI - Ephedrine in treatment of cardiac arrest. PMID- 17126474 TI - Modulation of involuntary attention by the duration of novel and pitch deviant sounds in children and adolescents. AB - In a passive auditory oddball event-related potential study, the processing of short (100 ms) and long (500 ms) novels and pitch deviant tones was investigated in three age groups (6-8, 10-12, and 17-18 years). Age specific distributions of P3a demonstrate developmental differences in the processing of unexpected sounds. Moreover, long compared with short novel sounds (but not long compared with short pitch deviant tones) elicited enhanced positive brain waves in early (200-300 ms) and late (300-400 ms) P3a as well as in post-P3a (400-600 ms) windows. This finding suggests stronger attentional capture for unexpected sounds with higher information content. The fact that in the post-P3a window this duration effect was largest for the 6-8 years old indicates that young children are especially prone for distraction. PMID- 17126475 TI - Short-term maternal and neonatal outcomes by mode of delivery. A case-controlled study. AB - OBJECTIVE: Side-by-side comparisons of short-term maternal and neonatal outcomes for spontaneous vaginal delivery, instrumental vaginal delivery, planned caesarean section and caesarean section during labor in patients matched for clinical condition, age, and week of gestation are lacking. This case-controlled study was undertaken to evaluate short-term maternal and neonatal complications in a healthy population at term by mode of delivery. STUDY DESIGN: Four groups of healthy women, with antenatally normal singleton pregnancies at term, who underwent instrumental vaginal delivery (no. 201), spontaneous delivery (no. 402), planned caesarean section without labor (no. 402) and caesarean section in labor (no. 402) have been retrospectively selected. Outcome measures were maternal and neonatal short-term complications. Odds ratios (OR) and 95% confidence intervals (CI) were calculated. RESULTS: Maternal complications were mostly associated with forceps-assisted and vacuum-assisted instrumental deliveries (OR: 6.9; 95% CI: 2.9-16.4 and OR 3.0; 95% CI 1.1-8.8, respectively, versus spontaneous deliveries). No significant differences in overall complications were observed between spontaneous vaginal deliveries and caesarean sections, whether planned or in labor. By comparison with caesarean sections in labor, instrumental deliveries significantly increased the risk of complications (OR: 3.2; 95% CI: 1.6-6.5). Neonatal complications were also mostly correlated with forceps-assisted and vacuum-assisted instrumental deliveries (OR: 3.5; 95% CI: 1.9-6.7 and OR 3.8; 95% CI 2.0-7.4, respectively, versus spontaneous deliveries). By comparison with caesarean sections in labor, instrumental vaginal deliveries significantly increased the risk of complications (OR: 4.2; 95% CI: 2.4-7.4). CONCLUSIONS: In healthy women with antenatally normal singleton pregnancies at term, instrumental deliveries are associated with the highest rate of short-term maternal and neonatal complications. PMID- 17126476 TI - Re: Weilie Hu, Jun Lu, Lichao Zhang, et al. A preliminary report of penile transplantation. Eur urol 2006;50:851-3. PMID- 17126477 TI - The detour extra-anatomic stent--a permanent solution for benign and malignant ureteric obstruction? AB - OBJECTIVES: We describe our experience of the Detour extra-anatomic stent (EAS) (Mentor-Porges, UK) for permanent bypass of complete upper urinary tract obstruction. The self-retaining expanded polytetrafluoroethylene-silicone tube, placed in the kidney using a percutaneous route, is tunnelled under the skin and sutured into the bladder to establish extra-anatomical urinary drainage. METHODS: From April 2002 to November 2005, a total of nine Detour stents were inserted into eight patients; one patient needed bilateral stent insertions. The causes for ureteric obstruction were persistent malignant disease in three and complicated benign disease in five patients. RESULTS: To date, four of five patients with benign disease are alive; one died unexpectedly of metastatic malignancy. The only stent-related complications were infection and haematuria. The two patients with malignancy have subsequently died, but there were no urinary drainage problems for their second and third years of life, respectively. CONCLUSIONS: The preliminary data presented here suggest that the Detour EAS offers a permanent and minimally invasive method to establish internalisation of urinary drainage to bypass complete ureteric obstructions for which conventional stenting has failed, open surgery has been tried and failed or was not considered feasible, and long-term nephrostomy drainage was not favoured. PMID- 17126478 TI - NeuroD1 expression in human prostate cancer: can it contribute to neuroendocrine differentiation comprehension? AB - OBJECTIVES: Neuroendocrine differentiation is a common feature of prostate cancer (pCA). NeuroD1 is a neuronal transcription factor able to convert epithelial cells into neurons. The aim of the study is to investigate NeuroD1 expression and compare it with chromogranin-A, synaptophysin, and CD56 staining in human prostate cell lines and surgical specimens. METHODS: We detected NeuroD1 gene expression, by duplex reverse transcriptase-polymerase chain reaction, in primary human prostate fibroblasts, in EPN, LNCaP, DU145, and PC3 cell lines before and after cAMP exposure, in 6 BPH and 11 pCA samples. Thereafter 166 paraffin sections from normal and neoplastic prostates were stained with NeuroD1, chromogranin-A, synaptophysin, and CD56 antibodies. The relationships between chromogranin-A and NeuroD1 and clinicopathologic parameters were evaluated by multivariate logistic regression analysis. RESULTS: NeuroD1 is inactive in baseline prostate cell lines and BPHs, whereas it is actively expressed in cAMP treated EPN, PC3, and DU145 cells. In our surgical series, positive chromogranin A, synaptophysin, CD56, and NeuroD1 staining was detected in 26.5%, 4.3%, 3.1%, and 35.5%, respectively (difference between chromogranin-A and NeuroD1: p<0.05). The multivariate analysis showed a strong association between chromogranin-A and microscopic perineural invasion (OR: 2.49; 95%CI, 0.85-7.32; p=0.097) and a high primary Gleason score (OR: 1.96; 95%CI, 1.14-3.39; p=0.015), whereas NeuroD1 expression strictly correlated to microscopic perineural invasion (OR: 2.97; 95%CI, 1.05-8.41; p=0.04). CONCLUSIONS: Expression of NeuroD1 versus chromogranin A is more frequent in pCA, and correlates to increased indicators of malignancy in moderately to poorly differentiated pCA, and could be involved in the pathophysiology of the neuroendocrine differentiation of pCA. PMID- 17126479 TI - Imaging of sellar and parasellar lesions. AB - The sellar and parasellar region is an anatomically complex area where a number of neoplastic, infectious, inflammatory, developmental and vascular pathologies can occur. Differentiation among various etiologies may not always be easy, since many of these lesions may mimic the clinical, endocrinologic and radiologic presentations of pituitary adenomas. The diagnosis of sellar lesions involves a multidisciplinary effort, and detailed endocrinologic, ophthalmologic and neurologic testing are essential. CT and, mainly, MRI are the imaging modalities to study and characterise normal anatomy and the majority of pathologic processes in this region. We here provide an overview of the most relevant MRI and CT characteristics together with clinical findings of pituitary tumors, vascular, inflammatory and infectious lesions found in the sellar/parasellar region in order to propose an appropriate differential diagnosis. PMID- 17126480 TI - Checklist for cognitive and emotional consequences following stroke (CLCE-24): development, usability and quality of the self-report version. AB - OBJECTIVE: In this paper a new checklist (CLCE-24) for identification of cognitive and emotional problems after stroke is presented. The CLCE-24 is intended to support a clinical interview by health care professionals other than the trained (neuro)psychologist. METHODS: Patients were interviewed with the CLCE 24, 6 months post stroke. Usability was determined by interviews. Quality of the self-report version was determined using reference instruments (MMSE, CAMCOG). RESULTS: Sixty-nine patients participated in the study (37 men; mean age 66 years). Both patients and assessors were positive about the use of the CLCE-24. Eighty percent of the patients had cognitive and/or emotional problems (73% cognitive; 51% emotional problems). Patients with complaints on the CLCE-24 also showed problems on the MMSE and the CAMCOG (p<0.05). The CLCE-24 was a predictor of the MMSE and CAMCOG (Adj. R(2)=0.13 and 0.16, respectively) at 12 months post stroke. Internal consistency of the CLCE-24 was good (alpha of 0.81). CONCLUSIONS: The CLCE-24 is a usable and valid instrument for cognitive screening by health care professionals in the stroke service in the chronic phase after stroke. PMID- 17126481 TI - Characteristics of equilibrium, kinetics studies for adsorption of fluoride on magnetic-chitosan particle. AB - The magnetic-chitosan particle was prepared and characterized by the SEM, XRD, FT IR and employed as an adsorbent for removal fluoride from the water solution in the batch system. The Langmuir isotherms, Bradley's isotherm, Freundlich isotherm and Dubinin-Kaganer-Radushkevich (DKR) isotherm were used to describe adsorption equilibrium. The kinetic process was investigated using the pseudo-first-order model, pseudo-second-order model and intra-particle diffusion model, respectively. The results show that the magnetic-chitosan particle is amorphous of irregular clumps in the surface with groups of RNH(2), RNH(3), Fe-O, etc. Bradley's equation and two-sites Langmuir isotherms were fitted well with the adsorption equilibrium data; the maximal amount of adsorption of 20.96-23.98 mg/l and free energy of 2.48 kJ/mol were obtained from the Bradley's equation, two sites Langmuir isotherm and DKR modeling, respectively. The pseudo-second-order with the initial adsorption rate 2.08 mg/g min was suitable to describe the kinetic process of fluoride adsorption onto the adsorbent. In overall, the major mechanism of fluoride adsorption onto the heterogeneous surface of magnetic chitosan particle was proposed in the study. PMID- 17126482 TI - Lessons learned from LNG safety research. AB - During the period from 1977 to 1989, the Lawrence Livermore National Laboratory (LLNL) conducted a liquefied gaseous fuels spill effects program under the sponsorship of the US Department of Energy, Department of Transportation, Gas Research Institute and others. The goal of this program was to develop and validate tools that could be used to predict the effects of a large liquefied gas spill through the execution of large scale field experiments and the development of computer models to make predictions for conditions under which tests could not be performed. Over the course of the program, three series of LNG spill experiments were performed to study cloud formation, dispersion, combustion and rapid phase transition (RPT) explosions. The purpose of this paper is to provide an overview of this program, the lessons learned from 12 years of research as well as some recommendations for the future. The general conclusion from this program is that cold, dense gas related phenomena can dominate the dispersion of a large volume, high release rate spill of LNG especially under low ambient wind speed and stable atmospheric conditions, and therefore, it is necessary to include a detailed and validated description of these phenomena in computer models to adequately predict the consequences of a release. Specific conclusions include: * LNG vapor clouds are lower and wider than trace gas clouds and tend to follow the downhill slope of terrain due to dampened vertical turbulence and gravity flow within the cloud. Under low wind speed, stable atmospheric conditions, a bifurcated, two lobed structure develops. * Navier-Stokes models provide the most complete description of LNG dispersion, while more highly parameterized Lagrangian models were found to be well suited to emergency response applications. * The measured heat flux from LNG vapor cloud burns exceeded levels necessary for third degree burns and were large enough to ignite most flammable materials. * RPTs are of two types, source generated and enrichment generated, and were observed to increase the burn area by a factor of two and to extend the downwind burn distance by 65%. Additional large scale experiments and model development are recommended. PMID- 17126483 TI - Solid phase extraction of chromium(VI) from aqueous solutions by adsorption of its ion-association complex with cetyltrimethylammoniumbromide on an alumina column. AB - A simple approach has been developed for the solid phase extraction of chromium(VI) based on the adsorption of its ion-association complex with cetyltrimethylammoniumbromide (CTABr) on an alumina column. The effect of various parameters such as acidity, stability of the column, sample volume, effect of diverse ions, etc. have been studied in detail. The adsorbed complex could be eluted using sodium hydroxide and the concentration of chromium has been established using visible spectrophotometry after complexation with diphenyl carbazide. The calibration graph was linear in the range 0-0.5 microg mL(-1) chromium(VI) with a detection limit of 5 microg L(-1). A highest preconcentration factor of 25 could be obtained for 250 mL sample volume. The data from the column studies were also studied using the Thomas model of adsorption. The experimental results obtained were correlated with the proposed model of adsorption. The Thomas rate constant k was found to be 0.0025 L/min mg and the maximum adsorption capacity q(o) was found to be 0.36 mgCr/g alumina at an initial chromium(VI) concentration of 1 mg L(-1). The validity of the method has been checked by applying it to study the recovery of chromium in spiked water samples and electroplating wastewater. PMID- 17126484 TI - Estimation of selected heavy metals and arsenic in PM10 aerosols in the ambient air of the Greater Athens Area, Greece. AB - Aerosol samples of PM(10) were collected during summer and winter 2003 at two different sites in the Messogia Basin northeast of Athens, to demonstrate the variations of heavy metals in PM(10) and examine their relationship with both gaseous pollutants and meteorological parameters. Estimated heavy metals during the experimental campaign were mercury (Hg), cadmium (Cd), lead (Pb), nickel (Ni) and arsenic (As). The average heavy metal concentrations for the first site (Spata) constituted 0.66-14.7ng/m(3) for the summer period and 0.14-19.5ng/m(3) for the winter period. At the second site (Koropi), the corresponding values varied between 0.89 and 13.3ng/m(3) and 0.16 and 24.7ng/m(3), respectively. PM(10) Hg, PM(10) Cd and PM(10) Ni contents showed regular daily variations, with higher mass percentages during the summer, indicating differences in local PM(10) sources for each season. On the contrary, PM(10) Pb presented higher mass percentages during the winter. Examination of the relationship between heavy metals and meteorological parameters indicated a higher correlation with temperature and relative humidity, especially for Pb. In addition, most of the heavy metals (apart from Hg) presented an expected correlation with nitrate oxides (NO(x)), PM(10) and ozone (O(3)). Higher correlations with both meteorological parameters and gaseous pollutants were observed during the winter experimental campaign. Maximum heavy metal concentrations at both sites were observed during days with NE or NNE prevailing winds during the summer campaign, while the winter period was characterized with maximums during days with W or WNW prevailing winds. PMID- 17126485 TI - The effect of acid treatment of carbon cloth on the adsorption of nitrite and nitrate ions. AB - The effects of functional groups on the adsorption efficiency of nitrate and nitrite from water by carbon cloth were investigated. The carbon cloth was treated by sulfuric acid and used for the adsorption of nitrate and nitrite from water samples at nearly neutral solutions. The concentrations of ions in the solution were monitored using in situ UV spectroscopy. Acid treatment caused a significant increase in the adsorption rate of ions and the adsorption capacity of the adsorbent. PMID- 17126486 TI - The reactivity of sodium borohydride with various species as characterized by adiabatic calorimetry. AB - The reactivity of sodium borohydride in the presence of other species has been examined by adiabatic calorimetry. In combination with water, sodium borohydride exhibits an exotherm at room temperature accompanied by generation of gas (presumed to be hydrogen). Addition of potassium hydroxide to a sodium borohydride-water mixture is found to stabilize the solution and require a higher temperature for reaction to occur. However, if iron oxide is also included, reaction takes place near room temperature. Very rapid reaction was found when a metal chloride was brought in contact with a solution containing sodium borohydride, water, and potassium hydroxide. When sodium borohydride was added to an oxygenated hydrocarbon, reaction at room temperature also took place, but to a more limited extent. Peak temperatures above 200 degrees C and maximum pressures in excess of 2000 psia were observed in most cases. Kinetics extracted from the calorimetry data are presented for some of the sodium borohydride combinations. PMID- 17126487 TI - Enhanced dissolution of TCE in NAPL by TCE-degrading bacteria in wetland soils. AB - The influence of trichloroethene (TCE) dechlorinating mixed cultures in dissolution of TCE in nonaqueous phase liquid (NAPL) via biodegradation was observed. Experiments were conducted in batch reactor system with and without marsh soils under 10 and 20 degrees C for 2 months. The dissolution phenomenon in biotic reactors containing mixed cultures was showed temporal increases compared to abiotic reactors treated with biocide. Effective NAPL-water transfer rate (K(m)) calculated in this study showed more than four times higher in biotic reactors than that in abiotic reactors. The results might be attributed to the biologically enhanced dissolution process via dechlorination in reactors. Temperature would be a factor to determine the dissolution rate by controlling bacterial activity. The TCE dechlorination occurred even in an interface of TCE NAPL that demonstrated no previous TCE biodegradation, suggesting that microbes may be useful in developing source-zone bioremediation system. In conclusion, dechlorinating mixed culture could enhance dissolution in NAPL that may be useful in the application of source zone bioremediation. PMID- 17126488 TI - The adsorption of chromium (VI) from industrial wastewater by acid and base activated lignocellulosic residues. AB - This study deals with the adsorption of Cr(VI) from synthetic and industrial wastewater, produced by a sewage plant. The activated carbons were prepared from a lignocellulosic raw material by thermal treatment at 450 and 650 degrees C in the presence of acid (AlCl(3), HCl, H(3)PO(4) and H(2)SO(4)) and base (NaOH) agents. To optimize the adsorption of Cr(VI), the chemical modifications caused by each activating agent (related to the capability of Cr(VI) removal), and the optimal experimental conditions of the pH, Cr(VI) concentration, adsorbent dose and residence time, were studied. Thus, treatment with H(3)PO(4) gives rise to carbons with a high surface area and high efficiency for Cr(VI) removal at short equilibrium times. In contrast, the generation of active surface sites by means of NaOH requires longer equilibrium times, the adsorption being less effective than in the former case. The adsorption isotherms obey the Langmuir equation only in the first stages of the reaction but fit the Freundlich equations over the whole range studied, so the heat of adsorption can be easily calculated. The results also show that the activated carbons obtained can be recovered by filtration with an efficiency of 30% in the third cycle. PMID- 17126489 TI - Many-beam dynamical simulation of electron backscatter diffraction patterns. AB - We present an approach for the simulation of complete electron backscatter diffraction (EBSD) patterns where the relative intensity distributions in the patterns are accurately reproduced. The Bloch wave theory is applied to describe the electron diffraction process. For the simulation of experimental patterns with a large field of view, a large number of reflecting planes has to be taken into account. This is made possible by the Bethe perturbation of weak reflections. Very good agreement is obtained for simulated and experimental patterns of gallium nitride GaN{0001} at 20kV electron energy. Experimental features like zone-axis fine structure and higher-order Laue zone rings are accurately reproduced. We discuss the influence of the diffraction of the incident beam in our experiment. PMID- 17126490 TI - Low-voltage cross-sectional EBIC for characterisation of GaN-based light emitting devices. AB - Electron beam induced current (EBIC) characterisation can provide detailed information on the influence of crystalline defects on the diffusion and recombination of minority carriers in semiconductors. New developments are required for GaN light emitting devices, which need a cross-sectional approach to provide access to their complex multi-layered structures. A sample preparation approach based on low-voltage Ar ion milling is proposed here and shown to produce a flat cross-section with very limited surface recombination, which enables low-voltage high resolution EBIC characterisation. Dark defects are observed in EBIC images and correlation with cathodoluminescence images identify them as threading dislocations. Emphasis is placed on one-dimensional quantification which is used to show that junction delineation with very good spatial resolution can be achieved, revealing significant roughening of this GaN p-n junction. Furthermore, longer minority carrier diffusion lengths along the c axis are found at dislocation sites, in both p-GaN and the multi-quantum well (MQW) region. This is attributed to gettering of point defects at threading dislocations in p-GaN and higher escape rate from quantum wells at dislocation sites in the MQW region, respectively. These developments show considerable promise for the use of low-voltage cross-sectional EBIC in the characterisation of point and extended defects in GaN-based devices and it is suggested that this technique will be particularly useful for degradation analysis. PMID- 17126491 TI - A simple method for linear burn debridement. AB - Debridement of linear burns especially on the trunk and abdomen can be challenging. Some currently used methods are technically difficult and potentially hazardous to the surgeon or the assistant. We propose a simple and effective method of linear burn debridement using towel clips. A pair of strong towel clips can be used to grasp the area of the burn at either end producing a tenting effect and therefore facilitating debridement of difficult areas. This technique provides a simple, precise and safe method of linear burn debridement. PMID- 17126492 TI - Adrenal androgen and gonadal hormone levels in adolescent girls with conduct disorder. AB - There are few data on the biological correlates of female antisocial behavior. This study compared adrenal androgen and gonadal hormone levels in adolescent girls with conduct disorder (CD) to girls without any psychiatric disorder (NC). We studied 87 girls, (47 CD; 36 NC), ages 15-17 years, obtaining three blood samples, drawn 20 min apart between 8 and 9 AM in the first 72 h of the onset of menstrual flow. Plasma was assayed for testosterone, estradiol, androstenedione, dehydroepiandrosterone (DHEA), dehydroepiandrosterone-sulfate (DHEA-S), sex hormone binding globulin (SHBG), and cortisol; area under the curve (AUC) for each of the three samples was used in the data analysis. We also calculated the Free Testosterone Index, Free Estrogen Index, Index of Hyperandrogenism and cortisol to DHEA ratio. In addition to receiving a full psychiatric interview, each girl completed a self-report questionnaire on general aggression. Main hormone analyses controlled for potentially confounding variables such as psychiatric comorbidity and race. Girls with CD had significantly lower cortisol to DHEA ratios, but did not differ from NC girls on any other hormone variable. Girls with symptoms of aggressive CD had significantly higher mean free testosterone indexes, lower SHBG levels, and lower cortisol to DHEA ratios than girls with non-aggressive CD. Girls with CD scored higher on the aggression questionnaire, but there was no association between general aggression and any hormone variable for the sample. Our data suggest that girls with CD, particularly aggressive CD, have lower cortisol to DHEA ratios, higher levels of free testosterone, and lower levels of SHBG. Clinical and research implications of these findings are discussed. PMID- 17126493 TI - Can pulmonary vein antrum be defined by electrophysiological mapping? AB - Atrial fibrillation (AF) is the most common sustained arrhythmia, and treatments with anti-arrhythmia drugs (AADs) have been frustrating. Limitations of AADs prompted the development of percutaneous catheter ablation. In contrast to AADs, percutaneous catheter ablation offers the possibility of a lasting cure. The successful cure of AF by percutaneous catheter ablation comes from a widespread recognize that pulmonary vein antrum (PVA) plays an important role in the genesis and maintenance of AF, and circular ablation along the PVA can eliminate majority of AF. PVA is comprised of pulmonary vein-left atrium junctions. However, during ablation procedure, definition of PVA solely depends on angiography, and it is largely experience-dependent and there is a great deal of variation involved. Our study in patients with AF found that a unique potential with double deflections could be documented along PVA, but it cannot be recorded at PV side or LA side. Thus, we propose that documentation of PVA potentials can be used as a landmark to define PVA. Unlike angiography, documentation of PVA potentials can be objectively carried out by different operators, and the variations due to experience can be avoided. PMID- 17126494 TI - On the mechanism of apoptosis-inducing activity of human calprotectin: zinc sequestration, induction of a signaling pathway, or something else? AB - Calprotectin, a heterodimer present in neutrophil cytoplasm, has antimicrobial and apoptosis-inducing activities. At the moment, there are two general hypotheses about the mechanism of action of calprotectin: (i) exclusion of extracellular zinc by calprotectin, and consequently induction of apoptosis; (ii) binding of calprotectin to a cell membrane receptor, and consequently, activation of a signaling pathway for apoptosis. Here, we introduce another hypothesis, i.e. inhibition or destruction of "target" inside cells. We suggest that calprotectin might become internalized non-specifically, maybe in a process like pinocytosis. This process is probably independent of the zinc concentration. We also demonstrated that the internal target hypothesis successfully predicts cell survival behavior of cultured cells as a function of calprotectin concentration. Additional analyses should be performed to elucidate the real calprotectin "target". PMID- 17126495 TI - Enhancement of endogenous defenses against ROS by supra-nutritional level of selenium is more safe and effective than antioxidant supplementation in reducing hypertensive target organ damage. AB - Hypertension-induced target organ damage (TOD), is one of the leading causes of morbidity and mortality. Reactive oxygen species (ROS) play an important role in the pathogenesis and development of hypertension. It has been suggested that hypertension-induced TOD is related to the level of oxidative stress, but is in part independent of the level of blood pressure. Therefore, in addition to anti hypertensive drug therapy, novel strategies against ROS, will provide additional benefits to patient with hypertension. Vitamin E has long been supplemented as an effective antioxidant. However, the potential hazardous effects of vitamin E supplementation as antioxidant revealed by recent studies make its clinical and routine use prudent. Therefore, novel approaches capable of enhancing endogenous system to defend against ROS are required. Here, we propose that enhancement of intrinsic defenses against ROS by supra-nutritional level of selenium is more safe and effective than antioxidant supplementation in reducing hypertensive target organ damage, owing to its role in activating and constitution of native vital proteins and/or enzymes against oxidative stress, and the fact that scarcity of selenium can not be supplemented by normal food, and potentially extra benefits by supra-normal intake. PMID- 17126496 TI - A novel TAT fusion protein with osteoinductive activity. AB - Osteoblasts are thought to be differentiated from pluripotent mesenchymal stem cells. Several intracellular and extracellular osteoinductive proteins are involved in this process. Such proteins include the bone morphogenetic proteins (BMPs) and the LIM mineralization proteins (LMPs) etc. LMP-1 is a novel LIM domain protein promoting the differentiation of osteoblasts during bone formation. It contains three LIM domains/motifs, one PDZ domain and a unique sequence. Through analysis of the amino acid sequence and the function of the LMPs, it has been found that the PDZ domain (1-93 aa) and a unique region (94-133 aa) appear to be critical for bone formation. The TAT protein of human immunodeficiency virus can be fused with other macromolecules, peptides or proteins and transport them into cells successfully. Once being transduced into cells, the fusion protein can recover its biological activity through being rapidly refolded. We supposed that TAT could be fused with LMP-1 (1-133 aa) and LMP-1 (94-133 aa) and the fusion proteins could be easily transduced through biological membranes and generate biological activity. The clinical application of BMPs has been limited for their relatively high cost and the unstable osteoinductivity. If the hypothesis proved to be practical, we would have a more effective new way to promote bone repair and regeneration. PMID- 17126497 TI - Insecticidal activity of Vitex mollis. AB - Organic extracts from the leaves of Vitex mollis were assessed for their toxic effect on fall armyworm neonate larvae (Spodoptera frugiperda), an important insect pest of corn. The extracts showed insecticidal and insect growth regulatory activity, being CHCl(3)-MeOH (1:1) extract the most active, also found to be toxic in the Artemia salina test. PMID- 17126498 TI - Characterization of two duplicate zebrafish Cb2-like cannabinoid receptors. AB - Several cannabinoid receptors have been detected in many organisms. The best known are CB1, mainly expressed in the central nervous system and CB2 which is almost exclusively expressed in the periphery. Here we report the molecular characterization of two duplicate CB2-like cannabinoid receptors from zebrafish (Danio rerio) (zebrafish Cb2a and zebrafish Cb2b). The amino acid sequences of these receptors present 56% identity with Takifugu rubripes CB2 sequence and 39% with human CB2 sequence and conserve some specific key residues for cannabinoid receptor function. Both duplicate receptors are expressed in peripheral tissues (gills, heart, intestine and muscle), immune tissue (spleen) and also in the central nervous system. Using in situ hybridization techniques zebrafish Cb2 mRNA expression was observed for the first time in the adenohypophysial cells of the rostral pars distalis and proximal pars distalis of the pituitary gland. Given the importance of the existence of duplication of genes in teleosts, the combined analysis of these two new cannabinoid receptors opens a new exciting door to investigate and understand cannabinoid function throughout evolution. PMID- 17126499 TI - Assembly of feline calicivirus-like particle and its immunogenicity. AB - Virus-like particles (VLPs) were produced in insect cells infected with a recombinant baculovirus containing the capsid gene of feline calicivirus strain F9 (FCV-F9). The FCV VLPs were morphologically and antigenically similar to the native virus and contained a single capsid protein with a molecular weight of approximately 60 kDa that reacted with FCV antiserum. Moreover, following immunization of rabbits, VLPs were able to elicit neutralizing antibodies against several FCV strains isolated from clinical samples. Our preliminary results showed that FCV-VLP could be considered a candidate vaccine against FCV infections. PMID- 17126500 TI - Clonal relatedness of Salmonella isolates associated with invasive infections in captive and wild-caught rattlesnakes. AB - This study examines the serotype distribution and clonal relatedness among Salmonella isolates obtained from healthy and diseased snakes. Isolates from extraintestinal body sites were obtained through routine diagnostic lab submissions from snakes in two facilities that had experienced a high prevalence of osteomyelitis in Crotalus species. Gastrointestinal isolates were predominantly from fecal samples collected from healthy snakes of both crotalid and non-crotalid species in one facility. PFGE macrorestriction analysis of Salmonella isolates confirmed the clonal and species-restricted nature of Salmonella serotype IIIa 56: z4, z23: - in one facility. Fourteen of 15 isolates from suspected osteomyelitis lesions in wild-caught snakes at the second facility were also from Salmonella subgroup IIIa (serotype IIIa 18: z4, z23: -) and appeared to be closely related by PFGE. Evaluation of a PCR assay for the spvC gene in 209 isolates demonstrated that this method consistently distinguished isolates of subgroup IIIa from those of subgroup IIIb. The data presented establish that Salmonella of subgroup IIIb are abundant and regularly associated with gastrointestinal shedding in snakes but that Salmonella in subgroup IIIa disproportionately cause infections in bone or other extraintestinal sites. PMID- 17126501 TI - Granulomatous lesions in lymph nodes of slaughter pigs bacteriologically negative for Mycobacterium avium subsp. avium and positive for Rhodococcus equi. AB - The prevalence of granulomatous lesions in lymph nodes of pigs was studied. From January till August 2004 in two slaughterhouses in The Netherlands 2,116,536 pigs were examined for the presence of granulomatous lesions in the sub-maxillary lymph nodes. In 15,900 (0.75%) of these pigs, lesions could be detected. Nine farms with the highest incidence of lesions were selected for a more detailed pathological and bacteriological examination. On these farms, the prevalence of lesions in sub-maxillary lymph nodes ranged from 2.3 to 5.7% with a mean of 3.0%. From 1276 pigs that were sampled, 98 (7.7%) displayed granulomatous lesions in the sub-maxillary lymph nodes and one (0.1%) pig showed lesions in its mesenteric lymph node. Mycobacterium avium subsp. avium (MAA) could not be isolated from the lymph nodes of the 99 pigs with lesions and from a selection of lymph nodes (n=61) of pigs without lesions. Rhodococcus equi was isolated from 44 out of 98 (44.9%) of the sub-maxillary lymph nodes with granulomatous lesions and from two mesenteric lymph nodes without lesions. A comparison of former studies and the current results indicate that the prevalence of MAA infections in slaughter pigs has strongly decreased over the last decade, whereas R. equi is highly prevalent. The high incidence of granulomatous lesions associated with the bacteriological presence of R. equi could be considered as a serious cause of misdiagnosis of MAA infections in cases where meat inspection is carried out by inspection for granulomatous changes of lymph nodes only. PMID- 17126502 TI - The expression of Clostridium perfringens consensus beta2 toxin is associated with bovine enterotoxaemia syndrome. AB - Clostridium perfringens has been implicated in a broad array of enteric infections including the fatal haemorrhagic enteritis/enterotoxaemia syndrome in cattle. The beta2 toxin (CPB2), encoded by cpb2, is suspected to be implicated in this syndrome. However, among C. perfringens isolates from cattle suspected of clostridial disease, an atypical allele was recently found to predominate at the cpb2 locus and atypical corresponding CPB2 proteins were shown to be poorly expressed, thus arguing against a biologically significant role of the beta2 toxin in clostridial diseases in cattle. This study compared genotype and phenotype of the beta2 toxin between C. perfringens isolates from a group of healthy calves (n=14, 87 isolates) and from a group of enterotoxaemic calves (n=8, 41 isolates). PCR results revealed the exclusive presence of the typical "consensus"cpb2 in the enterotoxaemic group. Western blot analysis demonstrated that the typical variant of CPB2 was often expressed in isolates from enterotoxaemic calves (43.9%) and infrequently in isolates from healthy cattle (6.9%). These data suggest that the typical variant of the CPB2 toxin may play a role in the pathogenesis of cattle enterotoxaemia. PMID- 17126503 TI - Cholinesterase pseudo-activity, oximolysis, esterolysis, thiocholine ester hydrolysis by oximes: what's in a name? PMID- 17126504 TI - Social rank and response to the "male effect" in the Australian Cashmere goat. AB - The present study was conducted to determine if the social status of Australian Cashmere goats affects their response to the male effect in terms of LH secretion, ovulation and expression of estrus. Australian Cashmere goats were kept isolated from the males during 5 months. The index of success (SI) of each goat was calculated to establish their social rank. In the first experiment, the ten most dominant and the 10 most subordinate goats were separated from the original herd and housed in two pens (5 dominant and 5 subordinate animals in each pen). An androgenized wether was then introduced into each pen. Luteinizing hormone (LH) was measured every 20 min from 2h before to 4h after introduction of the male in the goats of first pen and from 4 to 8h after male introduction in the second pen. In the second experiment, the remaining 50 goats were exposed in their original pen to two androgenized wethers. Their association index with the males (AI) was calculated for each of these 50 goats, and the intervals from exposure to the males to the onset of estrus and to ovulation were determined. During the first 4h after male introduction, the dominant goats had more LH pulses (0.65+/-0.06 compared with 0.3+/-0.09; P<0.05) and greater LH mean concentrations (1.79+/-0.14 ng/ml compared with 1.30+/-0.15 ng/ml, P=0.05) than the subordinate animals. Although not significantly different, the AI was 35% greater for high and medium ranking goats than for low ranking animals (0.031+/ 0.004, 0.032+/-0.005 and 0.023+/-0.005, respectively, P>0.05). Although the number of goats ovulating in response to male exposure was similar between dominance groups (high: 100%, medium: 94% and low ranking: 92%), the high and medium dominance goats showed a greater incidence of expression of estrus than low-dominance goats (94.4%, 89.5% and 53.8%, respectively, P<0.05). It is concluded that the social rank of the Australian Cashmere goat influences their response to the male effect in terms of early LH secretion and expression of estrus. PMID- 17126505 TI - Exogenous glucagon effects on health and reproductive performance of lactating dairy cows with mild fatty liver. AB - Severe fatty liver, a metabolic disease of dairy cows in early lactation, results in decreased health and reproductive performance, but can be alleviated by treatment with i.v. injections of glucagon. Mild fatty liver in cows effects on health and reproductive performance were determined by treatment with 14-day s.c. injections of glucagon at 7.5 or 15 mg/day. Multiparous Holstein cows (n=32) were grouped into Normal and Susceptible based on liver triacylglycerol concentrations (>1% liver tissue biopsy wet weight) at day 8 postpartum (day 0=day of parturition). Susceptible cows (n=24) were assigned randomly to three groups and s.c. injected with 0mg glucagon [60 ml 0.15M NaCl] [n=8] (same for Normal cows), 2.5 mg glucagon, or 5 mg glucagon every 8 h for 14 days, beginning day 8 postpartum. Mild fatty liver resulted in an increased number of days with elevated body temperature during the injection period, an increased incidence of mastitis after glucagon treatment, increased days to first estrus and insemination, increased days before conception occurred, and decreased conception rate. In cows with mild fatty liver, glucagon (15 mg/day) decreased the number of days with elevated body temperature and the incidence of mastitis after hormone treatment. From these results, we suggest that mild fatty liver is detrimental to health and reproduction of dairy cows and, furthermore, that exogenous glucagon decreases some of these detrimental effects. PMID- 17126506 TI - Studying thermodynamic aspects of sublimation, solubility and solvation processes and crystal structure analysis of some sulfonamides. AB - Crystal structures of N-(2-chlorophenyl)-benzene-sulfonamide (I), N-(2,3 dichlorophenyl)-benzene-sulfonamide (II), N-(4-chlorophenyl)-benzene-sulfonamide (III) were solved by X-ray diffraction method. Temperature dependencies of saturated vapor pressure and thermodynamic functions of sublimation process were calculated (I: DeltaG(sub)(298)=50.4kJmol(-1); DeltaH(sub)(298)=114+/-1kJmol(-1); DeltaS(sub)(298)=213+/-3Jmol(-1)K(-1); II: DeltaG(sub)(298)=54.1kJmol(-1); DeltaH(sub)(298)=124.9+/-1.6kJmol(-1); DeltaS(sub)(298)=237+/-5Jmol(-1)K(-1); III: DeltaG(sub)(298)=49.9kJmol(-1); DeltaH(sub)(298)=98.6+/-1.9kJmol(-1); DeltaS(sub)(298)=163+/-5Jmol(-1)K(-1)). Thermochemical parameters of fusion process for the compounds were obtained. Enthalpies of evaporation were estimated from enthalpies of sublimation and fusion. Temperature dependencies of the solubility in water, n-octanol and n-hexane were measured. The thermodynamic functions of solubility and solvation processes were deduced. Specific and non specific solvation terms were distinguished using the transfer from the "inert"n hexane to the other solvents. The transfer processes of the molecules from water to n-octanol were analyzed and main driven forces were established. PMID- 17126507 TI - Characterisation of salmon calcitonin in spray-dried powder for inhalation. Effect of chitosan. AB - Salmon calcitonin (sCT) powders suitable for inhalation, containing chitosan and mannitol as absorption enhancer and protection agent, respectively, were prepared using a spray-drying process. The effect of chitosan on physicochemical stability of sCT in the dry powder was investigated by different analytical techniques. High-performance liquid chromatography (HPLC) analysis indicated that sCT was chemically stable upon spray-drying. With the proportion of chitosan in spray drying formulation being increased, dissolution of sCT from the dry powders was decreased both in phosphate buffer and acetate buffer. The thioflavine T fluorescence assay showed that no fibrils were present in the spray-dried powder. However, sCT partly fibrillated in the phosphate buffer, but not in acetate buffer. Fourier transform infrared (FTIR) spectra showed that the secondary structure of sCT was slightly changed in the dry powder, yet no aggregate signal was observed. Circular dichroism analysis indicated that the structure of sCT in an aqueous formulation was slightly altered by addition of chitosan. Nevertheless, recovery of sCT was not influenced by chitosan in the aqueous formulation as indicated by HPLC analysis. This study suggested that sCT, in absence of any additives, was stable during the spray-drying process under certain conditions. Addition of chitosan affects recovery of sCT from spray-dried powders, which may be due to formation of a partially irreversible complex between the protein and chitosan during the spray-drying process. PMID- 17126508 TI - Antiplasmodial and GABA(A)-benzodiazepine receptor binding activities of five plants used in traditional medicine in Mali, West Africa. AB - Extracts of five medicinal plants: Boscia angustifolia, Cissus quadrangularis, Securidaca longipedunculata, Stylosanthes erecta and Trichilia emetica, used traditionally in Malian traditional medicine were screened for in vitro antiplasmodial activity and GABA(A)-benzodiazepine receptor binding activity. Four extracts showed significant antiplasmodial activities, with the dichloromethane extract of leaf of Securidaca longipedunculata being the most active (IC(50) of 7 microg/ml [95% CI: 5-9]). The dichloromethane extract of leaf of Trichilia emetica, in addition to its antiplasmodial activity (IC(50): 12 microg/ml [95% CI: 12-14]), exhibited a good binding activity to the GABA(A) benzodiazepine receptor, while water and methanol extracts of the same plant did not show any activity. A strong GABA(A)-receptor complex binding activity was observed in the methanol extract of aerial part of Stylosanthes erecta. The results in this study justify some of the traditional indications of the plants investigated and may thus be candidates for Improved Traditional Medicines in Mali. PMID- 17126509 TI - Effect of Brazilian green propolis on experimental gastric ulcers in rats. AB - Propolis is a resinous hive product collected by honeybees from plants. The propolis produced in Southeastern of Brazil is known as green propolis because of its color. Modern herbalists recommend its use because it displays antibacterial, antifungal, antiviral, hepatoprotective, anti-inflammatory, immunomodulatory and anti-ulcer properties. The anti-ulcer activity of green propolis hydroalcoholic crude extract was evaluated by using models of acute gastric lesions induced by ethanol, indomethacin and stress in rats. Moreover, the effects of extract on gastric content volume, pH and total acidity, using pylorus ligated model were evaluated. Animals pretreated with propolis hydroalcoholic crude extract (50, 250 and 500 mg/kg) showed a significant reduction in lesion index, total affected area and percentage of lesion in comparison with control group (p<0.05) in the ethanol-induced ulcer model. Green propolis extract, at a higher dose (500 mg/kg), displayed a significant protection by reducing (p<0.05) the evaluated parameters in the gastric ulceration induced by indomethacin. In the stress induced ulcer model it was observed a significant reduction (p<0.05) in those parameters in animals treated with green propolis extract (250 and 500 mg/kg). Regarding the pylorus ligated model it was observed that green propolis extract (250 and 500 mg/kg) displayed an anti-secretory activity, which lead to a reduction in the gastric juice volume, total acidity and pH. These findings indicate that Brazilian green propolis displays good anti-ulcer activity, corroborating the folk use of propolis preparations, and contributing for its pharmacological validation. PMID- 17126510 TI - Death associated with volatile substance inhalation--histologic, scanning electron microscopic and energy dispersive X-ray spectral analyses of lung tissue. AB - The investigation of deaths due to the inhalation of volatile substances may be complicated by a lack of scene and autopsy findings. Mechanisms of death may not be determinable at autopsy, and there may be very few markers of inhalant abuse. A 21-year-old man is reported who died from the combined effects of methadone toxicity and toluene inhalation. Histological examination of the lungs revealed congestion and edema, as well as particles of blue, pigmented material within the interstitium and in macrophages. Scanning electron microscopy was undertaken, revealing that the particles contained granules that measured 0.15-0.2microm in diameter, within the range of mean particle sizes for inorganic paint pigments. Energy dispersive X-ray spectral analysis of the granules demonstrated a significant percentage of titanium (12%) confirming their origin from paint. Ancillary investigations such as electron microscopy and X-ray spectral analysis in cases of possible lethal volatile inhalation may prove useful adjuncts in determining the type of substance inhaled and in providing evidence of previous non-lethal episodes. PMID- 17126511 TI - Drosophila aging 2005/06. AB - Drosophila continues to be a model system of choice to study the genetics of aging. It has a short lifespan and small genome size, but nevertheless contains a complex organ and endocrine system that allows studying the role of conserved signal transduction pathways with sophisticated genetic tools. Oxidative stress and metabolic changes along with intersecting signaling systems Insulin Receptor (InR), Target of Rapamycin (TOR) and Jun N-terminal Kinase (JNK) have emerged as some of the major players in aging. Sleep and organ-specific aging has also been the subject of recent progress in understanding aging. PMID- 17126512 TI - Yeast as a model for chronological and reproductive aging - a comparison. PMID- 17126513 TI - Aging: a revisited theory based on free radicals generated by NOX family NADPH oxidases. AB - Reactive oxygen species (ROS), often also referred to as free radicals, play an important role in aging. It is widely assumed that mitochondria are the predominant source of ROS relevant for the aging process. In this hypothesis article, we suggest that the role of ROS generated by NOX family NADPH oxidases has been largely overlooked in aging theories. NOX NADPH oxidases form a seven member gene family and are high level ROS-generating enzymes. As opposed to mitochondria, which generate ROS as a byproduct of their metabolism, NOX enzymes are professional ROS generators. From an experimental point of view, there is now abundant evidence for the involvement of NOX enzymes in age-associated diseases. The role of NOX enzymes in the aging process itself and their relative contribution as compared to mitochondria needs further investigations. PMID- 17126514 TI - Delayed neurodegeneration and early astrogliosis after excitotoxicity to the aged brain. AB - Excitotoxicity is well recognised as a mechanism underlying neuronal cell death in several brain injuries. To investigate age-dependent differences in neurodegeneration, edema formation and astrogliosis, intrastriatal N-methyl-d aspartate injections were performed in young (3 months) and aged (22-24 months) male Wistar rats. Animals were sacrificed at different times between 12h and 14 days post-lesion (DPL) and cryostat sections were processed for Toluidine blue, Fluoro-Jade B staining, NeuN and GFAP immunohistochemistry. Our results show that both size of tissue injury and edema were reduced in the old subjects only up to 1DPL, correlating with a slower progression of neurodegeneration with peak numbers of degenerating neurons at 3DPL in the aged, contrasting with maximum neurodegeneration at 1DPL in the young. However, old animals showed an earlier onset of astroglial response, seen at 1DPL, and a larger area of astrogliosis at all time-points studied, including a greater glial scar. In conclusion, after excitotoxic striatal damage, progression of neurodegeneration is delayed in the aged but the astroglial response is earlier and exacerbated. Our results emphasize the importance of using aged animals and several survival times for the study of acute age-related brain insults. PMID- 17126515 TI - Protective effect of an extract of the oyster mushroom, Pleurotus ostreatus, on antioxidants of major organs of aged rats. AB - This study was undertaken to investigate the putative antioxidant activity of the oyster mushroom, Pleurotus ostreatus, on lipid peroxidation and antioxidant status of major organs of aged (24 month old) rats when compared to young (4 month old) rats. Elevated levels of malondialdehyde (MDA) and significantly lowered levels of reduced glutathione (GSH) and of vitamins C and E were observed in the liver, kidneys, heart and brain of aged rats, when compared to values in young rats. Quantitative analysis of the activities of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (Gpx) revealed significantly lower values in the liver, kidneys, heart and brain of aged rats. An analysis of isozyme pattern of these enzymes in aged rats also revealed variations in relative concentration, presumably due to oxidative stress. Administration of the extract of P. ostreatus to aged rats resulted in elevated levels of reduced glutathione and vitamins C and E and in increased activities of CAT, SOD and Gpx so that the values in most of these parameters did not differ significantly from those in young rats. In addition, the level of MDA was lowered on administration of mushroom extract to aged rats. These results suggest that treatment with an extract of P. ostreatus can improve the antioxidant status during ageing, therein minimizing the occurrence of age associated disorders associated with involvement of free radicals. PMID- 17126517 TI - Degradation product of loratadine. PMID- 17126516 TI - Oxidative stress, mitochondria and mtDNA-mutator mice. AB - The oxidative stress theory of aging, an expansion of the mitochondrial theory of aging, is based around the idea of a vicious cycle, in which somatic mutations of mitochondrial DNA (mtDNA) provoke respiratory chain dysfunction leading to enhanced ROS production and in turn to the accumulation of further mtDNA mutations. Mitochondrial dysfunction and mtDNA mutations are amplified during the course of aging. Recently, results obtained from mtDNA-mutator mice further strengthen the role of mitochondria in the aging process. However, lack of increased oxidative stress in the mtDNA-mutator mice raises doubts in the direct connection of mtDNA mutations with increased ROS production, challenging the oxidative stress theory of aging. The purpose of this short review is to highlight several studies that provide direct evidence that accelerated aging is linked to mtDNA mutations, without an increase in oxidative damage. PMID- 17126518 TI - Isolation and characterization of degradation impurities in docetaxel drug substance and its formulation. AB - The degradation of docetaxel drug substance and its injection formulation has been investigated. The majority of impurities were observed in a base degradation study and all five degradation products were characterized. These impurities were isolated, enriched and were subjected to mass and NMR spectral studies. Based on the spectral data, these were characterized as 10-deacetyl baccatin III, 7-epi-10 deacetyl baccatin III, 7-epi-10-oxo-10-deacetyl baccatin III, 7-epi docetaxel and 7-epi-10-oxo-docetaxel, respectively. The last two impurities were also detected in the stability study of docetaxel formulation. Out of these degradation impurities two substances have been previously identified while the other three previously unreported. PMID- 17126519 TI - Simultaneous determination of loratadine and desloratadine in pharmaceutical preparations using liquid chromatography with a microemulsion as eluent. AB - A rapid HPLC procedure for analytical quality control of pharmaceutical preparations containing the antihistaminic drug substance loratadine and/or its analog desloratadine (which is also an active metabolite of loratadine) was developed using a microemulsion as the eluent. The separation was performed on a column packed with cyanopropyl bonded stationary phase adopting UV detection at 247 nm using a flow rate of 1 ml/min. The optimized microemulsion mobile phase consisted of 0.1M sodium dodecyl sulphate, 1% octanol, 10% n-propanol and 0.3% triethylamine in 0.02 M phosphoric acid, pH 3.0. The developed method was validated in terms of specificity, linearity, lower limit of quantification, lower limit of detection, precision and accuracy. With the proposed method satisfactory resolution between loratadine and desloratadine (resolution factor=3.85). The method requires a minimum of sample handling and is rapid (10 min), and reproducible (R.S.D.<2.0%). The mean recoveries of the analytes in pharmaceutical preparations were in agreement with those obtained from a reference method, as revealed by statistical analysis of the obtained results using the Student's t-test and the variance ratio F-test. Pseudoephedrine, the co formulated drug substance, did not interference with the assay and was successfully separated using the developed HPLC method. PMID- 17126520 TI - Randomized comparison of percutaneous Viabahn stent grafts vs prosthetic femoral popliteal bypass in the treatment of superficial femoral arterial occlusive disease. AB - OBJECTIVE: This randomized prospective study was designed to compare the effectiveness of treating superficial femoral artery occlusive disease percutaneously with expanded polytetrafluoroethylene (ePTFE)/nitinol self expanding stent grafts vs surgical femoral-to-above knee (AK) popliteal artery bypass with synthetic graft material. METHODS: From March 2004 to May 2005, 100 limbs in 86 patients with femoral-popliteal arterial occlusive disease were identified. Patients had symptoms ranging from claudication to rest pain, with or without tissue loss, and were prospectively randomized for treatment into one of two groups. The limbs were treated percutaneously with angioplasty and one or more self-expanding stent grafts (n = 50) or surgically with femoral-to-AK popliteal artery bypass using synthetic Dacron or ePTFE grafts (n = 50). The mean +/- SD total length of artery stented was 25.6 +/- 15 cm. Follow-up evaluation with ankle-brachial indices and color flow duplex sonography imaging were performed at 3, 6, 9, and 12 months after treatment. RESULTS: Patients were monitored for a median of 18 months. No statistical difference was found in the primary patency (P = .895) or secondary patency (P = .861) between the two treatment groups. Primary patency at 3, 6, 9, and 12 months of follow-up was 84%, 82%, 75.6%, and 73.5% for the stent graft group and 90%, 81.8%, 79.7%, and 74.2% for the femoral-popliteal surgical group. Thirteen patients in the stent graft group had 14 reinterventions, and 12 reinterventions occurred in the surgical group. This resulted in secondary patency rates of 83.9% for the stent graft group and 83.7% for the surgical group at the 12-month follow-up. CONCLUSIONS: Management of femoral-popliteal arterial occlusive disease using percutaneous treatment with a stent graft is comparable with surgical revascularization with conventional femoral-to-AK popliteal artery bypass using synthetic material up to 12 months. Longer-term follow-up would be helpful in determining ongoing efficacy. PMID- 17126522 TI - The role of 18F-fluoro-deoxy-glucose positron emission tomography (FDG-PET) in the management of patients with colorectal cancer. AB - AIM: In patients with colorectal cancer an accurate diagnostic work-up is mandatory in order to perform the most specific treatment. At this moment 18F fluoro-deoxy-glucose positron emission tomography (FDG-PET) is considered an accurate imaging technique in staging/restaging several malignancies. The aim of this paper is to review the scientific literature available about the role of FDG PET in the management of patients with colorectal cancer. METHODS: An overview on Medline of scientific literature concerning FDG-PET and colorectal cancer was performed. The most relevant studies are reported. Advantages, limitations and new chances in using FDG-PET in these subsets of patients are summarized. RESULTS: FDG-PET is a useful tool in the evaluation of colorectal cancer. In comparison to conventional imaging technique, FDG-PET has an additional diagnostic value because it allows to metabolically characterize undetermined lesions suspected for recurrence of disease, to perform a complete pre-surgical staging and to identify occult metastatic disease. In clinical practice its use leads to a change in therapeutic choices in a high percentage of cases. CONCLUSIONS: FDG-PET should be considered an essential diagnostic tool in the management of patients with colorectal cancer, especially in recurrent disease evaluation. PMID- 17126523 TI - Hereditary diffuse gastric cancer and E-cadherin: description of the first germline mutation in an Italian family. AB - AIMS: Germline mutation of the E-cadherin gene (CDH1) accounts for the Hereditary Diffuse Gastric Cancer (HDGC) syndrome. Fourteen pedigrees with Diffuse Gastric Cancer that fulfilled the International Gastric Cancer Linkage Consortium (IGCLC) criteria were selected and screened for CDH1 germline mutations. METHODS: The entire coding region of the CDH1 gene and all intron-exon boundaries were analyzed by direct sequencing in the 14 families fulfilling the IGCLC criteria. E cadherin immunohistochemical expression was evaluated on tumour as well as normal formalin-fixed paraffin embedded tissues. RESULTS: A novel germline missense mutation was found. It was a single C-->T substitution in exon 8, resulting in a transition of CCG-->CTG (C1118T; Pro373Leu) demonstrated in the proband and her brother. At immunohistochemical analysis, the staining intensity was reduced and considered weakly positive (15%). CONCLUSIONS: The first CDH1 germline mutation of an Italian family is herein reported. The present missense mutation has never been described so far. PMID- 17126524 TI - (99M)Technetium-sestamibi scintimammography in non-palpable breast lesions found on screening X-ray mammography. AB - OBJECTIVE: The sensitivity and specificity of (99m)Tc-sestamibi scintimammography in patients with non-palpable breast lesions diagnosed by screening mammography and the value of (99m)Tc-sestamibi to detect axillary lymph node metastases was determined. METHODS: Between September 2000 and December 2003, 103 females with non-palpable breast lesions were included for further evaluation. X-ray mammography was repeated and 99mTc-sestamibi scintimammography performed within one-week. Anterior, and left and right lateral images were obtained. The scintimammography was analysed by 2 experienced observers who were blinded to the clinical, pathological, and radiological results. The sensitivity and specificity of scintimammography to diagnose non-palpable lesion(s), including the axillary regions, was compared with histopathology, clinical, and radiological follow up. RESULTS: Two patients (one non-small lung cancer and one non-Hodgkin's disease) were excluded. Both showed (99m)Tc-sestamibi avid lesions in the breast and axillary region. In the remaining 101 patients, 37 true positive (TP), 4 false positive (FP), 52 true negative (TN), and 8 false negative (FN) breast carcinomas were found. The specificity was 92.8%, sensitivity 82.2%, positive predictive value (PPV) 90.2%, and negative predictive value (NPV) 86.6%. (99m)Tc-sestamibi scintimammography showed axillary lesions in 5/15 (33%) patients with axillary lymph node metastasis. CONCLUSION: In patients with non-palpable lesions diagnosed by screening- X-ray-mammography, (99m)Tc-sestamibi scintimammography provided high specificity and PPV. Furthermore, (99m)Tc-sestamibi scintimammography detected 33% of patients with axillary lymph node metastases. Therefore, (99m)Tc-sestamibi scintimammography could be of incremental value in the surgical work-up of these patients. PMID- 17126525 TI - An improved in vitro model to characterize invasive growing cancer cells simultaneously by function and genetic aberrations. AB - Invasion into the surrounding tissue and bone metastasis is a common feature of advanced prostate cancer. Chromosomal and other genetic or epigenetic abnormalities were aligned to this behaviour mostly by using permanent cell lines, paraffin embedded tissue or primary tumour samples. Both attempts fail to reflect either the original situation or functional information in the patient's tissue. Thus, we developed an improved in vitro assay to follow invasion of prostate cancer cells derived from fresh samples of radical prostatectomy specimens. Fresh tumour samples were applied onto Matrigeltrade mark-coated invasion chambers using a cocultivation model. Invasive growing cells were harvested from the bottom of the membrane or from the underlying gel and further characterized using comparative genomic hybridization. Prostate cancer cells have the capability to invasively grow through the barrier of a Matrigeltrade mark and could easily be sampled in a pad of Matrigeltrade mark. Comparative genomic hybridization revealed characteristic chromosomal aberrations of the invasive growing cells. Noteworthy is their ability to spheroid formation, which allows for further cell propagation by standard cell culture methods. Thus, our improved invasion model is a tool for the sampling of invasive growing cancer cells from fresh human tumour material allowing for functional as well as genetic studies. PMID- 17126526 TI - Allele-specific histone modifications regulate expression of the Dlk1-Gtl2 imprinted domain. AB - Dlk1 and Gtl2 are reciprocally expressed imprinted genes located on mouse chromosome 12. The Dlk1-Gtl2 locus carries three differentially methylated regions (DMRs), which are methylated only on the paternal allele. Of these, the intergenic (IG) DMR, located 12 kb upstream of Gtl2, is required for proper imprinting of linked genes on the maternal chromosome, while the Gtl2 DMR, located across the promoter of the Gtl2 gene, is implicated in imprinting on both parental chromosomes. In addition to DNA methylation, modification of histone proteins is also an important regulator of imprinted gene expression. Chromatin immunoprecipitation was therefore used to examine the pattern of histone modifications across the IG and Gtl2 DMRs. The data show maternal-specific histone acetylation at the Gtl2 DMR, but not at the IG DMR. In contrast, only low levels of histone methylation were observed throughout the region, and there was no difference between the two parental alleles. An existing mouse line carrying a deletion/insertion upstream of Gtl2 is unable to imprint the Dlk1-Gtl2 locus properly and demonstrates loss of allele-specific methylation at the Gtl2 DMR. Further analysis of these animals now shows that the loss of allele-specific methylation is accompanied by increased paternal histone acetylation at the Gtl2 DMR, with the activated paternal allele adopting a maternal acetylation pattern. These data indicate that interactions between DNA methylation and histone acetylation are involved in regulating the imprinting of the Dlk1-Gtl2 locus. PMID- 17126527 TI - Differential effects of neonatal handling on early life infection-induced alterations in cognition in adulthood. AB - We have previously demonstrated that bacterial infection (Escherichia coli) in neonatal rats is associated with impaired memory in a fear-conditioning task in adulthood. This impairment, however, is only observed if a peripheral immune challenge (lipopolysaccharide; LPS) is administered around the time of learning. We used a brief separation/handling paradigm to determine if the adult memory impairment associated with neonatal-infection could be prevented. Naturally occurring variations in maternal care promote striking variations in offspring cognitive development, and handling paradigms are used to manipulate the quality and quantity of maternal care. Rats were injected on post natal (P) day 4 with E. coli or PBS, and half from each group were handled for 15 min/day from P4 to 20. All rats were then tested in adulthood. Neonatal handling of rats infected as neonates prevented the increase in microglial cell marker reactivity within the hippocampus, and the exaggerated brain IL-1beta production to LPS normally produced by the infection. Thus, these neural processes were now comparable to levels of non-infected PBS controls. Furthermore, handling completely prevented LPS-induced memory impairment in a context-fear task in adult rats infected as neonates. Finally, neonatal handling dramatically improved spatial learning and memory and decreased anxiety in rats treated early with PBS, but had no beneficial effect on these measures in rats infected as neonates. Taken together, these data suggest that maternal care may profoundly influence neuroinflammatory processes in adulthood, and that infection may also prevent maternal care influences on cognition later in life. PMID- 17126528 TI - Illness-induced anorexia and its possible function in the caterpillar, Manduca sexta. AB - Although many animals exhibit illness-induced anorexia when immune-challenged, the adaptive significance of this behavior remains unclear. Injecting Manduca sexta larvae (caterpillars) with live bacteria (Serratia marcescens), heat-killed bacteria or bacterial lipopolysaccharides resulted in a decline in feeding, demonstrating illness-induced anorexia in this species. We used M. sexta to test four commonly suggested adaptive functions for illness-induced anorexia. (1) Food deprivation did not reduce the iron content of the hemolymph. (2) Immune challenged M. sexta were not more likely to move to a different part of the plant. Therefore, the decline in feeding is unlikely to be an adaptive response allowing the animal to move away from a patch of contaminated food. (3) M. sexta force-fed S. marcescens bacteria were not more susceptible to a S. marcescens systemic infection than were M. sexta force-fed nutrient broth. (4) Force-feeding infected M. sexta during illness-induced anorexia did not increase mortality and short-term food deprivation did not enhance survival. However, force-feeding M. sexta with a high lipid diet (linseed oil and water) resulted in an increase in mortality when challenged with S. marcescens. Force-feeding sucrose or water did not reduce resistance. Force-feeding a high lipid diet into healthy animals did not reduce weight gain, suggesting that it was not toxic. We hypothesize that there is a conflict between lipid metabolism and immune function, although whether this conflict has played a role in the evolution of illness-induced anorexia remains unknown. The adaptive function of illness-induced anorexia requires further study in both vertebrates and invertebrates. PMID- 17126529 TI - RGSZ1 interacts with protein kinase C interacting protein PKCI-1 and modulates mu opioid receptor signaling. AB - Protein kinase C interacting protein (PKCI-1) was identified among the potential interactors from a yeast two hybrid screen of human brain library using N terminal of RGSZ1 as a bait. The cysteine string region, unique to the RZ subfamily, contributes to the observed interaction because PKCI-1 interacted with N-terminus of RGS17 and GAIP, but not with that of RGS2 or RGS7 where cysteine string motif is absent. The interaction between RGSZ1 and PKCI-1 was confirmed by coimmunoprecipitation and immunofluorescence. PKCI-1 and RGSZ1 could be detected by coimmunoprecipitation using 14-3-3 antibody in cells transfected with PKCI-1 or RGSZ1 respectively, but when transfected with PKCI-1 and RGSZ1 together, only RGSZ1 could be detected. Phosphorylation of Galphaz by protein kinase C (PKC) reduces the ability of the RGS to effectively function as GTPase accelerating protein for Galphaz, and interferes with ability of Galphaz to interact with betagamma complex. We investigated the roles of 14-3-3 and PKCI-1 in phosphorylation of Galphaz. Phosphorylation of Galphaz by PKC was inhibited by 14 3-3 and the presence of PKCI-1 did not provide any further inhibition. PKCI-1 interacts with mu opioid receptor and suppresses receptor desensitization and PKC related mu opioid receptor phosphorylation [W. Guang, H. Wang, T. Su, I.B. Weinstein, J.B. Wang, Mol. Pharmacol. 66 (2004) 1285.]. Previous studies have also shown that mu opioid receptor co-precipitates with RGSZ1 and influence mu receptor signaling by acting as effector antagonists [J. Garzon, M. Rodriguez Munoz, P. Sanchez-Blazquez, Neuropharmacology 48 (2005) 853., J. Garzon, M. Rodriguez-Munoz, A. Lopez-Fando, P. Sanchez-Blazquez Neuropsychopharmacology 30 (2005) 1632.]. Inhibition of cAMP by mu opioid receptor was significantly reduced by RGSZ1 and this effect was enhanced in combination with PKCI-1. Our studies thus provide a link between the previous observations mentioned above and indicate that the major function of PKCI-1 is to modulate mu opioid receptor signaling pathway along with RGSZ1, rather than directly mediating the Galphaz RGSZ1 interaction. PMID- 17126530 TI - Regulation of IL-21 signaling by suppressor of cytokine signaling-1 (SOCS1) in CD8(+) T lymphocytes. AB - Mice lacking the gene for suppressor of cytokine signaling 1 (SOCS1) show defective homeostasis of T lymphocytes due to accumulation of CD8(+) T cells, resulting at least partly from dysregulated IL-15 signaling. IL-15 alone does not stimulate proliferation of naive CD8 T cells, but can synergize with IL-21 to induce proliferation, suggesting a potential role for IL-21 in the defective homeostasis of CD8(+) T lymphocytes in SOCS1(-/-) mice. Since IL-21 strongly induced SOCS1 mRNA in CD8(+) T cells, we investigated whether SOCS1 regulates their response to IL-21. CD8(+) T cells isolated from SOCS1-deficient mice proliferated vigorously in response to IL-21+IL-15. In CD8(+) T lymphocytes expressing transgenic TCR, IL-21+IL-7 provided a stronger stimulus to naive cells whereas IL-15+IL-21 potently stimulated memory cells. Compared to truly naive or memory cells, SOCS1(-/-) H-Y TCR(+) CD8(+) T cells displayed CD44(lo)Ly6C(hi)CD122(int)CD127(lo) partial memory phenotype and exhibited stronger response to IL-15+IL-21 than truly naive cells. In SOCS1(-/-) CD8(+) T cells, IL-21 caused greater reduction in IL-15 threshold for activation in a dose dependent manner. SOCS1 deficiency did not modulate IL-21Ralpha expression or sensitivity to IL-21, but delayed the loss of IL-21-induced phospho-STAT3 signal. These results show that SOCS1 is a critical regulator of IL-21 signaling in CD8(+) T cells, and support the notion that sustained IL-21 signaling might also contribute to the aberrant T cell homeostasis in SOCS1-deficient mice. PMID- 17126531 TI - Pharmacology and mechanism of action of pregabalin: the calcium channel alpha2 delta (alpha2-delta) subunit as a target for antiepileptic drug discovery. AB - Pregabalin (Lyrica) is a new antiepileptic drug that is active in animal seizure models. Pregabalin is approved in US and Europe for adjunctive therapy of partial seizures in adults, and also has been approved for the treatment of pain from diabetic neuropathy or post-herpetic neuralgia in adults. Recently, it has been approved for treatment of anxiety disorders in Europe. Pregabalin is structurally related to the antiepileptic drug gabapentin and the site of action of both drugs is similar, the alpha2-delta (alpha2-delta) protein, an auxiliary subunit of voltage-gated calcium channels. Pregabalin subtly reduces the synaptic release of several neurotransmitters, apparently by binding to alpha2-delta subunits, and possibly accounting for its actions in vivo to reduce neuronal excitability and seizures. Several studies indicate that the pharmacology of pregabalin requires binding to alpha2-delta subunits, including structure-activity analyses of compounds binding to alpha2-delta subunits and pharmacology in mice deficient in binding at the alpha2-delta Type 1 protein. The preclinical findings to date are consistent with a mechanism that may entail reduction of abnormal neuronal excitability through reduced neurotransmitter release. This review addresses the preclinical pharmacology of pregabalin, and also the biology of the high affinity binding site, and presumed site of action. PMID- 17126532 TI - The uncinate fasciculus and extraversion in schizotypal personality disorder: a diffusion tensor imaging study. PMID- 17126533 TI - Positive association between SIAT8B and schizophrenia in the Chinese Han population. AB - The Sialyltransferase 8B gene (SIAT8B) is located at 15q26, a susceptibility region for both schizophrenia and bipolar disorder. The protein encoded by this gene has an important role in neural development and sialic acid synthesis on the neural cell adhesion molecule (NCAM). Previous research had indicated that the promoter region of SIAT8B is associated with schizophrenia in the Japanese population. To take this further we carried out an association study based on 643 unrelated schizophrenics and 527 unrelated healthy subjects, all Han Chinese, recruited from Shanghai. Although our results differed from those of the Japanese research, rs3759915, also located in the promoter region of SIAT8B, showed nominally significant association with schizophrenia (P=0.0036). Moreover, haplotypes constructed from rs3759915 and another two SNPs reported in the Japanese study (rs3759914 and rs3759916, also located in promoter region of SIAT8B) which located in the same LD block were significantly associated with schizophrenia (global P=0.0000050). Our findings indicate that SIAT8B may be a candidate susceptibility gene for schizophrenia in the Chinese Han population and may also provide further support for the potential importance of polysaccharide synthesizing genes in the etiology of schizophrenia. PMID- 17126534 TI - Intraventricular or intrathecal use of polymyxins in patients with Gram-negative meningitis: a systematic review of the available evidence. AB - Several reports have described the use of polymyxins by the intraventricular or intrathecal route for multidrug-resistant Gram-negative meningitis. We reviewed the available clinical evidence regarding intraventricular/intrathecal administration of polymyxins in patients with meningitis, focusing on effectiveness and safety. Relevant studies were identified from PubMed (January 1950 to April 2006) as well as from the references of relevant articles. We identified 31 case reports/series that matched our inclusion criteria. Sixty-four episodes of Gram-negative meningitis (34 in adults) were reviewed. Monotherapy with polymyxins via the intraventricular or intrathecal route was used in 11 episodes and combination of systemic and local polymyxins was used in 25 episodes. In the remaining episodes, various combinations of local polymyxins with systemic and/or local antibiotics were administered. Cure was achieved in 51/64 episodes (80%); in 26/30 episodes (87%) due to Pseudomonas aeruginosa and in 10/11 episodes (91%) due to Acinetobacter spp. Toxicity related to local administration of polymyxins was noted in 17/60 (28%) patients. The most common toxicity was meningeal irritation (12 cases). Discontinuation of treatment was necessary in four episodes and dose reduction in four episodes; irreversible toxicity was not reported. The limited available evidence suggests that therapy with intraventricular/intrathecal polymyxins alone or in combination with systemic antimicrobial agents is effective against Gram-negative meningitis. Toxicity is not uncommon but it is dose-dependent and reversible. Further studies are needed to evaluate the criteria for initiation of local central nervous system treatment with polymyxins, the optimal dosages and the role of adjuvant systemic or local therapy. PMID- 17126535 TI - Immunological evaluation of SW-HSA conjugate on goats. AB - Locoweeds cause significant livestock poisoning and economic loss all over the world. The purpose of this study was to investigate the immune effects of locoweed toxin, swainsonine (SW) and human serum albumin (HSA) conjugate (SW HSA), on goats. Twenty-four Sannon goats were randomly separated into immune control group (eight goats), immune poisoning group I (six goats), immune poisoning group II (six goats) and poisoning control group (four goats). Immune control group, immune poisoning groups I and II were first vaccinated with SW-HSA conjugate. The poisoning control group, immune poisoning groups I and II were then fed with 10.0 g/kg BW/day dry powder of Oxytropis kansuensis Bunge everyday morning. The immune control group was supplied with an alfalfa-based diet. Blood samples of these experimental animals were collected at different time interval. Immunoassay was performed using indirect ELISA and E-rosette technique. The results show that, after second booster immunization: (1) anti-SW antibody level in some goats increased to 2(8), which proves that SW-HSA conjugate can induce experimental animals to produce high-level anti-SW antibody in their bodies; (2) the high-level antibody in their bodies could maintain 30 days, and decreased gradually after poisoning experiment (in our experiment, there was a return of the antibody level on day 21 after poisoning experiment); (3) the decreasing of the E-rosette rate of the immune poisoning group was delayed 14 days, which suggests that SW-HSA could low down the loss of the immunity of the goats; (4) swainsonine concentration in the blood was significantly lower (p<0.01) in the immune poisoning groups than that in the poisoning control group, and there was no significant difference (p>0.01) between the two immune poisoning groups within the poisoning experiment. PMID- 17126536 TI - Static light scattering study of complex formation between protein and neutral water-soluble polymer. AB - Complex formation of poly(N-isopropylacrylamide) (PNIPA) having a weight-average molecular weight of 1,720,000g/mol with human serum albumin (HSA), ovalbumin (OVA) and lysozyme (LYZ) was studied in an aqueous medium containing 0.01 M NaCl and adjusted to pH 3. The polymer-protein mixtures at different molar ratios (r(m)) were examined by static light scattering (SLS). The analysis of SLS data using our own approach [Kokufuta et al., Langmuir 15 (1999) 940; Biomacromolecules 4 (2003) 728] showed that the molecular weight of each resulting complex is smaller than that of the interpolymer complex composed of two polymer chains plus one protein. This indicates the formation of an intrapolymer complex in all the polymer-protein systems studied. Thus, at each r(m) we calculated the number of bound proteins per polymer, the value of which was OVA>HSA>LYZ in order. These results were compared with the hydropathy profiles of each protein which are a good tool for obtaining an information about distribution of hydrophobic and hydrophilic segments in a protein. It has become apparent that the hydrophobic interaction between polymer and protein plays an important role in the intrapolymer complex formation. PMID- 17126537 TI - Colloidal crystallization of colloidal silica modified with ferrocenyl group contained polymers in organic solvents. AB - Surface modification of colloidal silica with ferrocenyl-grafted polymer and colloidal crystallization of the particles in organic solvent were studied. Poly(methyl methacrylate-co-vinylferrocene)-grafted silica never formed colloidal crystals in polar solvent, such as acetone, acetonitrile, ethanol and N,N dimethylformamide (DMF), while poly(methyl methacrylate-co-ferrocenyl acrylate) grafted silica gave colloidal crystallization in DMF. The particles prepared by grafting of poly(N,N-dimethylacrylamide-co-vinylferrocene), with vinylferrocene (Vfc) mole fraction of 1/13 and 1/23, were observed to give the crystallization in ethanol and DMF over particle volume fraction of 0.058. Further, silica modified with copolymer of Vfc and N-vinyl-2-pyrrolidone, N-vinylcarbazole or N isopropylacrylamide formed colloidal crystals in ethanol and DMF. Especially, poly(N-isopropylacrylamide-co-Vfc)-grafted silica, which was composed of the highest mole fraction of vinylferrocene, 1/3, afforded colloidal crystallization in ethanol over particle volume fraction of 0.053. Relatively high polar vinylferrocene copolymer grafting of silica resulted in colloidal polymerization in organic solvents. PMID- 17126538 TI - Drying dissipative patterns of the colloidal crystals of silica spheres in an dc electric field. AB - Drying dissipative structural patterns of the colloidal crystals of silica spheres were studied under an dc-electric field. Platinum plate electrodes of anode and cathode were set on a cover glass. The broad hills accumulated with the spheres were observed at the outer edges of the dried film without and also with the electric fields. The column-like structures were formed by the electric flux, and movement of the spheres took place toward anode. The dried film kept colloidal crystal structure, where the nearest-neighbored spheres contact each other more compactly in the areas closer to the anode. Drying times needed for the complete dryness of the suspensions decreased as the strength of the electric field increased. Addition of sodium chloride to the suspensions retarded the movement of spheres toward the anode substantially. PMID- 17126539 TI - Fine structure and putative feeding mechanism of the archigregarine Selenidium orientale (Apicomplexa: Gregarinomorpha). AB - Archigregarines are considered one of the most plesiomorphic groups of Apicomplexa. Until recently, however, this viewpoint was based mainly on the results of the detailed investigation of a single species, Selenidium hollandei. The present study of the fine structure of trophozoites of another archigregarine species Selenidium orientale Bogolepova, 1953. (Apicomplexa, Archigregarinida, as proposed by Grasse and Schrevel), with special reference to the forebody structure of the attached individuals, allows more confident discussion of the plesiomorphic status of the archigregarines. Specimens of S. orientale were collected from the midgut of the Pacific sipunculid Themiste pyroides Chamberlain, 1920. The ultrastructure of the trophozoites generally corresponds to that of other studied species of Selenidium. Differences in the forebody structure between S. orientale and S. hollandei do not conflict with Schrevel's hypothesis on the feeding function of the apical complex in archigregarine trophozoites, although they suggest that, in S. orientale at least, the cytostome is not a persistent structure, but re-opens for each sucking event, and Selenidium trophozoites feed by intermittent sucking of host cytoplasm. Microtubules in the axial zone of the mucron neck may mediate the transport of food vacuoles. PMID- 17126540 TI - Innate recognition of intracellular bacteria. AB - The molecular repertoire for innate recognition of bacterial pathogens has expanded rapidly in the past decade. These immunosensors include Toll-like receptors and the more recently defined NOD-like receptors (NLRs): NODs, NALPs, NAIP and IPAF. Toll-like receptors signal from the cell surface or endosome upon ligand binding, whereas NLRs are activated by characteristic bacterially derived molecules, such as peptidoglycan, RNA, toxins and flagellin, in the cytosol. Studies using animal and culture models of bacterial infection indicate a pro inflammatory role for NLRs, mediated by signaling through nuclear transcription factor kappaB and activation of caspase-1 by the inflammasome. These data also support a synergistic role for extracellular and intracellular bacterial sensing in regulating inflammation. In humans, NLR mutations are often associated with autoinflammatory syndromes, suggesting a complex role for cytosolic surveillance in systemic innate immunity. PMID- 17126541 TI - New and emerging roles for mast cells in host defence. AB - Mast cells are highly effective sentinel cells, found close to blood vessels and especially common sites of potential infection, such as the skin, airways and gastrointestinal tract. Mast cells participate actively in the innate immune responses to many pathogens through a broad spectrum of mediators that can be selectively generated. They also have a role as innate effector cells in enhancing the earliest processes in the development of acquired immune responses. Studies of bacterial and parasitic models have revealed mast cell dependent regulation of effector cell recruitment, mucosal barrier function and lymph node hypertrophy. An important role for mast cells in viral infection is also implied by several in vivo and in vitro studies. There are multiple direct and indirect pathways by which mast cells can be selectively activated by pathogens including Toll-like receptors, co-receptors and complement component receptors. Understanding the mechanisms and scope of the contribution of mast cells to host defence will be crucial to regulating their activity therapeutically. PMID- 17126542 TI - Horizontal transfer and hypovirulence associated with double-stranded RNA in Beauveria bassiana. AB - Beauveria bassiana strains from different hosts and geographic origins were assayed for the presence of double-stranded RNA (dsRNA). Two of them (15.4%) showed extra bands, with approximately 4.0-3.5 kb and 2-0.7 kb, respectively, after electrophoretic separation of undigested nucleic acids. Virus-like particles were approximately 28-30 nm diam. The dsRNA was maintained after conidiogenesis (vertical transmission) and was transmitted horizontally by hyphal anastomosis. Strains purged of dsRNA obtained after cycloheximide treatment showed increased conidial production when compared with strains carrying dsRNA particles. Bioassays demonstrated hypovirulence associated with dsRNA. The mean mortality against the insect Euschistus heros was reduced in strains containing dsRNA when compared with the isogenic dsRNA-free ones. PMID- 17126543 TI - Outcome of non-invasive domiciliary ventilation in elderly patients. AB - STUDY OBJECTIVES: To analyze the short- and long-term effects of domiciliary non invasive ventilation (NIV) in the elderly. METHODS: From 1990 to 2005 all patients who initiated NIV at age 75 or older were included in the study. The mean follow-up period was 36 (24) months. Data were obtained from a database record. RESULTS: Forty-three patients, mean age 77 (1.9) years and hypercapnic respiratory failure secondary to restrictive, neuromuscular or hypoventilatory disease were included. The short-term effects included a significant improvement in arterial blood gases and nocturnal desaturations during NIV compared to baseline: PaO(2) increased a mean of 19 mmHg (P<0.0001), PaCO(2) decreased a mean of 16 mmHg (P<0.0001) and nocturnal time with SaO(2)<90% decreased a mean of 72% (P<0.0001). Arterial blood gases while breathing room air also improved significantly at 6 months after NIV initiation. Five patients (11%) discontinued treatment; this group did not differ from patients who continued NIV. Mean compliance was 8.3 (3.1)h/day. In the long-term effects, we observed that the initial improvement of arterial blood gases breathing room air was maintained throughout the followup period. The number of hospital admissions and days of hospital stay decreased significantly (P<0.0001 and 0.001, respectively) after NIV initiation. The poorest survival was observed in ALS patients (median 10.9 (2.3) months) significantly lower than the survival for the other diagnostic groups (median 58.5 (4.8) months), P=0.0013. CONCLUSIONS: NIV is an effective treatment in the elderly. It improves arterial blood gases and nocturnal desaturations, decreases hospital admissions and is associated with long survival. So advanced age should not be considered as an exclusion criteria to prescribe NIV. PMID- 17126544 TI - Electrochemical magneto immunosensing of antibiotic residues in milk. AB - A novel electrochemical immunosensing strategy for the detection of sulfonamide antibiotics in milk based on magnetic beads is presented. Among the different strategies for immobilizing the class-specific anti-sulfonamide antibody to the magnetic beads--such as those based on the use of Protein A or carboxylate modified magnetic beads - ,the best strategy was found to be the covalent bonding on tosyl-activated magnetic beads. The immunological reaction for the detection of sulfonamide antibiotics performed on the magnetic bead is based on a direct competitive assay using a tracer with HRP peroxidase for the enzymatic labelling. After the immunochemical reactions, the modified magnetic beads can be easily captured by a magneto sensor made of graphite-epoxy composite (m-GEC), which is also used as the transducer for the electrochemical immunosensing. The electrochemical detection is thus achieved through a suitable substrate for the enzyme HRP and an electrochemical mediator. The electrochemical approach is also compared with a novel magneto-ELISA with optical detection. The performance of the electrochemical immunosensing strategy based on magnetic beads was successfully evaluated using spiked milk samples, and the detection limit was found to be 1.44 microg L(-1) (5.92 nmol L(-1)) for raw full cream milk. This strategy offers great promise for rapid, simple, cost-effective and on-site analysis of biological, food and environmental samples. PMID- 17126545 TI - Emerging concepts in molecular MRI. AB - Molecular magnetic resonance imaging (MRI) offers the potential to image some events at the cellular and subcellular level and many significant advances have recently been witnessed in this field. The introduction of targeted MR contrast agents has enabled the imaging of sparsely expressed biological targets in vivo. Furthermore, high-throughput screens of nanoparticle libraries have identified nanoparticles that act as novel contrast agents and which can be targeted with enhanced diagnostic specificity and range. Another class of magnetic nanoparticles have also been designed to image dynamic events; these act as 'switches' and could be used in vitro, and potentially in vivo, as biosensors. Other specialized MR probes have been developed to image enzyme activity in vivo. Lastly, the use of chemical exchange and off-resonance techniques have been developed, adding another dimension to the broad capabilities of molecular MRI and offering the potential of multispectral imaging. These and other advances in molecular MRI offer great promise for the future and have significant potential for clinical translation. PMID- 17126546 TI - Change from continuous epidural infusion to patient-controlled epidural analgesia on the labour ward of a large district general hospital. PMID- 17126547 TI - Treatment of brewery slurry in thermophilic anaerobic sequencing batch reactor. AB - Treatment of brewery slurry in a thermophilic anaerobic sequencing batch reactor (ASBR) was studied using conventional fully mixed semi-continuous digestion as a control. The process phases were adapted to fit the brewery slurry discharge schedule. ASBR experiments were conducted under different organic loading rates (OLR) from 3.23 to 8.57 kg of COD/m(3)day of reactor and control was conducted with OLR of 3.0 kg of COD/m(3)day. The ASBR COD degradation efficiency was from 79.6% to 88.9%, control experiment efficiency was 65%. ASBR VSS removal efficiency was from 78.5% to 90.5%, control experiment efficiency was 54%. The ASBR methane production yield was from 371 to 418 L/kg COD inserted, control experiment methane yield was 248 L/kg COD inserted. The ASBR process was superior to conventional fully mixed digestion, and is fully adaptable to brewery slurry discharge, needs no additional collection and settling pools and experiences no solids settling problems. PMID- 17126548 TI - A conserved mechanism of Hedgehog gradient formation by lipid modifications. AB - Members of the Hedgehog (Hh) family of proteins are conserved morphogens that modulate cell fates in target tissues in different developmental systems. Dysregulation of Hh signaling results in a wide range of human diseases. The mature Hh is modified by lipids in two places, with palmitate at the N-terminus and cholesterol at the C-terminus. The lipid modifications are essential to the proper secretion and spreading of the morphogen throughout the extracellular matrix, interacting with heparan sulfate proteoglycans. However, the role of lipid modifications in regulating the range and activity of Hh proteins remains controversial. Here, we aim to resolve this issue by providing a model that is consistent with current and past reports. We propose that the cholesterol moiety functions to restrict the dilution and deregulated spread of the morphogen in the extracellular space. PMID- 17126549 TI - Catching a GEF by its tail. AB - The activation of Rho GTPases is mediated by guanine-nucleotide exchange factors (GEFs), which catalyze the exchange of GDP for GTP. Rho-GEFs are a very diverse family, with >70 members in humans. Bioinformatics analysis of the human Rho-GEFs shows that approximately 40% contain a putative PDZ-binding motif at the C terminus. PDZ domains are protein-protein interaction domains that act as scaffolds to concentrate signaling molecules at specialized regions in the cell. We propose that the interaction between Rho-GEFs and PDZ-domain proteins is a general mechanism that controls Rho-GEF targeting and activation, helping to restrict and concentrate the exchange activity to appropriate subcellular destinations. Here, we summarize recent data that highlight the importance of these interactions in Rho-GEF regulation. PMID- 17126550 TI - Mitochondrial ROS--radical detoxification, mediated by protein kinase D. AB - The mitochondrial electron transport chain is the major source for the production of oxygen radicals. Mitochondria-generated reactive oxygen species (mROS) have been implicated in decreasing the life span and contributing to age-related diseases (known as the free radical theory of aging). Recently, the serine/threonine kinase protein kinase D1 (PKD1) was identified as a mitochondrial sensor for oxidative stress. mROS-activated PKD regulates a radical sensing signaling pathway, which relays mROS production to the induction of nuclear genes that mediate cellular detoxification and survival. This PKD regulated signaling pathway is the first known mitochondria located and mitochondrially regulated antioxidant system that protects these organelles and cells from oxidative stress-mediated damage or cell death. The identification of this and further intracellular protective signaling pathways provides an opportunity to manipulate the effects of mROS, and might provide the key to targeting aging effects and age-related diseases that have been linked to mitochondrial dysfunctions. PMID- 17126551 TI - A new system for rapid plasmid integration in Plasmodium parasites. AB - Transfection of the human malaria parasite Plasmodium falciparum has facilitated greater understanding of the biology of this devastating protozoal pathogen. However, technical limitations have restricted the options available for functional analysis. A recent study by Nkrumah and colleagues provides a powerful new transfection tool, the Bxb1 integrase system. In this article, we outline the potential of this system, describing how it enables direct site-specific integration and the rapid generation of stably transformed populations that express uniform levels of introduced transgenes. PMID- 17126552 TI - Subarachnoid clot distribution in anterior wall saccular aneurysms of the internal carotid artery. AB - Anterior wall aneurysms of the internal carotid artery (AWAICA) are relatively uncommon. We investigated the distribution of the subarachnoid clot on computerized tomography (CT) scans in patients with ruptured AWAICA and compared the findings with those in patients with internal carotid artery aneurysms (ICAA) at other sites. Twenty-six ruptured ICAA patients were included in this study. Four (15.4%) of these had an AWAICA and 22 had an ICAA at other sites. Three of the 4 patients with AWAICA had an ordinary 'saccular type' aneurysm and the other had a 'blood blister-like' aneurysm. In all three patients with a 'saccular type' AWAICA, accumulation of the subarachnoid clot in the olfactory sulcus was noted on CT scan. In the patient with a 'blood blister-like' aneurysm and in 22 patients with ICAA at other sites, there were no specific correlations between focal accumulations of the subarachnoid clot and aneurysm location. The accumulation of subarachnoid clot in the olfactory sulcus on CT scan may suggest a ruptured 'saccular type' AWAICA. PMID- 17126553 TI - Picrosirius-polarization staining method as an efficient histopathological tool for collagenolysis detection in vesical prolapse lesions. AB - The Picrosirius-polarization method has been indicated as a selective histochemical stain for collagen detection in tissue sections. This method can also be of value for studying collagen degradation given that, under polarized light, collagen displays birefringence due to its molecular order. The aim of this study is to highlight this staining method as an additional instrument for a rapid and excellent confirmatory diagnosis of the presence of collagenolysis in connective tissue in the vaginal wall with vesical prolapse lesion, in tissue sections. Dramatic changes in collagen morphology were found in vaginal mucosa in vesical prolapse disorder: they were weakly stained by Sirius red and under polarized light appeared as thin, pale (weakly birefringent), greenish, and with fibers more scattered, while the histoarchitecture of the organ showed a disrupted appearance. Thus, in the present study, we showed in vaginal mucosa in the vesicle prolapse that corroded collagenous framework appears as fragmentary and irregularly separated collagenous structures, that are weakly birefringent, corresponding to a molecular disorganization of these fibers caused by collagenolysis. PMID- 17126554 TI - Cardiac dysfunction in the R6/2 mouse model of Huntington's disease. AB - Recent evidence suggests that mutant huntingtin protein-induced energetic perturbations contribute to neuronal dysfunction in Huntington's disease (HD). Given the ubiquitous expression of huntingtin, other cell types with high energetic burden may be at risk for HD-related dysfunction. Early-onset cardiovascular disease is the second leading cause of death in HD patients; a direct role for mutant huntingtin in this phenomenon remains unevaluated. Here we tested the hypothesis that expression of mutant huntingtin is sufficient to induce cardiac dysfunction, using a well-described transgenic model of HD (line R6/2). R6/2 mice developed cardiac dysfunction by 8 weeks of age, progressing to severe failure at 12 weeks, assessed by echocardiography. Limited evidence of cardiac remodeling (e.g. hypertrophy, fibrosis, apoptosis, beta(1) adrenergic receptor downregulation) was observed. Immunogold electron microscopy demonstrated significant elevations in nuclear and mitochondrial polyglutamine presence in the R6/2 myocyte. Significant alterations in mitochondrial ultrastructure were seen, consistent with metabolic stress. Increased cardiac lysine acetylation and protein nitration were observed and were each significantly associated with impairments in cardiac performance. These data demonstrate that mutant huntingtin expression has potent cardiotoxic effects; cardiac failure may be a significant complication of this important experimental model of HD. Investigation of the potential cardiotropic effects of mutant huntingtin in humans may be warranted. PMID- 17126555 TI - One-stage total repair of aortic arch anomaly using regional perfusion. AB - OBJECTIVE: Primary repair of aortic arch obstructions and associated cardiac anomalies is a surgical challenge in neonates and infants. Deep hypothermic circulatory arrest prolongs myocardial ischemia and might induce cerebral and myocardial dysfunction. METHODS: From March 2000 to December 2005, 69 neonates or infants with aortic arch anomaly underwent one-stage biventricular repair with continuous cerebral perfusion in the presence of a nonworking beating heart using the dual perfusion technique on the innominate artery and aortic root. Preoperative diagnoses of arch anomaly comprised aortic coarctation (n=54) or an interrupted aortic arch (n=15). Combined anomalies were ventricular septal defect (n=52), anomalous origin of the right pulmonary artery from ascending aorta (n=3), hypoplastic left heart syndrome (n=2), truncus arteriosus (n=2), atrioventricular septal defect (n=2), double outlet right ventricle (n=1), total anomalous pulmonary venous return (n=1), partial anomalous pulmonary venous return (n=1), and aortic stenosis (n=1). RESULTS: The mean regional perfusion time was 27.8+/-9.8 min. There was no operative mortality. Postoperative low cardiac output was present in four patients (5.8%). A neurologic complication was noted in one patient (1.5%) who developed transient chorea, but recovered completely. During 32.8+/-17.5 months of follow-up, one late death (1.5%) occurred. There was neither reoperation associated with arch anomaly nor recoarctation except in one patient. One patient developed left main bronchial compression necessitating aortopexy. CONCLUSIONS: One-stage total arch repair using our regional perfusion technique is an excellent method that may minimize neurologic and myocardial complications without mortality. Our surgical strategy for arch anomaly has a low rate of residual and recurrent coarctation when performed in neonates and infants. PMID- 17126556 TI - Morbidity, mortality, and long-term survival after sleeve lobectomy for non-small cell lung cancer. AB - OBJECTIVE: Sleeve lobectomy is a widely accepted procedure for central tumors for which the alternative is pneumonectomy. The purpose of this study is to assess operative mortality, morbidity, and long-term results of sleeve lobectomies performed for non-small cell lung carcinoma (NSCLC). METHODS: A retrospective review of 218 patients who underwent sleeve lobectomy for NSCLC between 1981 and 2005 was undertaken. There were 186 (85%) men and 32 women with a mean age of 61.9 years (range, 19-82 years). Eighty patients (36.6%) had a preoperative contraindication to pneumonectomy. Right upper lobectomy was the most common operation (45.4%). Vascular sleeve resection was performed in 28 patients (12.8%) and was commonly associated with left upper lobectomy (n=20; 9.1%; p=0.0001). The histologic type was predominantly squamous cell carcinoma (n=164; 75%), followed by adenocarcinoma (n=46; 21%). Resection was incomplete in nine (4.1%) patients. RESULTS: There were nine operative deaths; the operative mortality and the morbidity rates were 4.1% and 22.9%, respectively. A total of 14 (6.4%) patients presented with bronchial anastomotic complications: two were fatal postoperatively, seven patients required reoperation, three required a stent insertion, and two were managed conservatively. Multivariate analysis showed that compromised patients (p=0.001), current smoking (p=0.01), right sided resections (p=0.003), bilobectomy (p=0.03), squamous cell carcinoma (p=0.03), and presence of N1 or N2 disease (p=0.01) were risk factors for mortality and morbidity. Follow-up was complete in 208 patients (95.4%). Overall 5-year and 10-year survival rates were 53% and 28.6%, respectively. After complete resection, recurrence was local in 10 patients, mediastinal in 20, and distant in 25. By multivariate analysis, two factors significantly and independently influenced survival: nodal status (N0-N1 vs N2; p=0.01) and the stage of the lung cancer (stage I-II vs III, p=0.02). CONCLUSIONS: For patients with NSCLC, sleeve lobectomy achieves local tumor control, even in patients with preoperative contraindication to pneumonectomy and is associated with low mortality and bronchial anastomotic complication rates. Postoperative complications are higher in compromised patients, smokers, N disease, right sided resections, bilobectomies, and squamous cell cancers. The presence of N2 disease and stage III significantly worsen the prognosis. PMID- 17126557 TI - Geometric models of the aortic and pulmonary roots: suggestions for the Ross procedure. AB - OBJECTIVE: To discuss geometric factors, which may influence long-term results relating to homograft competence following the Ross procedure, we describe the 3D morphology of the pulmonary and aortic roots. MATERIALS: Measurements were made on 25 human aortic and pulmonary roots. Inter-commissural distances and the heights of the sinuses were measured. For geometrical reconstruction the three commissures and their vertical projections at the root base were used as reference points. RESULTS: In the pulmonary root, the three inter-commissural distances were of similar dimensions (17.9+/-1.6mm, 17.5+/-1.4mm and 18.6+/ 1.5mm). In the aortic root, the right inter-commissural distance was greatest (18.8+/-1.9mm), followed by the non-coronary (17.4+/-2.0mm) and left coronary sinus commissures (15.2+/-1.9mm). The mean height of the left pulmonary sinus was greatest (20+/-1.7mm) followed by the anterior (17.5+/-1.4mm) and right pulmonary sinus (18+/-1.66mm). In the aortic root, the height of the right coronary sinus was the greatest (19.4+/-1.9mm) followed by the heights of the non-coronary (17.7+/-1.8mm) and left coronary sinus (17.4+/-1.4mm). Measured differences between parameters determine the tilt angle and direction of the root vector. The tilt angle in the pulmonary root averaged 16.26 degrees , respectively; for the aortic roots, it was 5.47 degrees . CONCLUSIONS: Herein we suggest that the left pulmonary sinus is best implanted in the position of the right coronary sinus, the anterior pulmonary in the position of the non-coronary sinus and the right pulmonary sinus in the position of the left coronary sinus. In this way, the direction of the pulmonary root vector will be parallel to that of the aortic root vector. PMID- 17126558 TI - Six-month outcome of transapical transcatheter aortic valve implantation in the initial seven patients. AB - BACKGROUND: The current treatment of choice for symptomatic aortic stenosis is aortic valve replacement (AVR) with cardiopulmonary bypass (CPB), but AVR is associated with significant operative morbidity and mortality in elderly patients with multiple co-morbid conditions. We recently reported the first successful aortic valve implantation procedure (AVI) via a mini-thoracotomy and left ventricular apical puncture without cardiopulmonary bypass. We now report 6-month follow-up in our initial seven patients. METHODS: Seven patients (77+/-10 years old) with symptomatic aortic stenosis were deemed to be non-surgical candidates for AVR and not suitable for a transfemoral percutaneous heart valve implantation due to aorto-iliac disease. The predicted 30-day operative mortality was 31+/-23% according to logistic Euroscore. Patients underwent minimally invasive transapical AVI. With the guidance of fluoroscopy and transesophageal echocardiography, balloon predilation was followed by deployment of a 26mm Cribier-Edwardstrade mark valve (Edwards Lifesciences Inc., Irvine, CA) during rapid ventricular pacing to reduce forward flow and cardiac motion. RESULTS: Valve implantation was successful in all seven patients. There were no intra procedural mortalities or complications. Thirty-day operative mortality was 14%. One patient died at day 12 due to pneumonia. Two patients died from non-cardiac diseases at day 51 and 85. The remaining four patients completed 6-month follow up. The aortic valve area increased from 0.7+/-0.3 to 1.8+/-0.7 and 1.5+/ 0.5cm(2) at 1 and 6 months, respectively. The mean transaortic gradient was reduced from 32+/-8 to 10+/-5 and 11+/-8mmHg at 1 and 6 months, respectively. Following AVI, none or trivial, mild, and moderate aortic regurgitation was observed in 4, 2, and 1 patients, respectively. There were no valve-related complications during the follow-up. CONCLUSION: Aortic valve implantation can successfully be performed via a minimally invasive apical approach without the need for cardiopulmonary bypass. The early results in this initial series are encouraging. This initial experience suggests that the minimally invasive transapical approach is a viable alternative for patients in whom open-heart surgery is not feasible or poses unacceptable risks. PMID- 17126559 TI - A strategy to search for common obesity and type 2 diabetes genes. AB - Worldwide, the incidence of type 2 diabetes is rising rapidly, mainly because of the increase in the incidence of obesity, which is an important risk factor for this condition. Both obesity and type 2 diabetes are complex genetic traits but they also share some nongenetic risk factors. Hence, it is tempting to speculate that the susceptibility to type 2 diabetes and obesity might also partly be due to shared genes. By comparing all of the published genome scans for type 2 diabetes and obesity, five overlapping chromosomal regions for both diseases (encompassing 612 candidate genes) have been identified. By analysing these five susceptibility loci for type 2 diabetes and obesity, using six freely available bioinformatics tools for disease gene identification, 27 functional candidate genes have been pinpointed that are involved in eating behaviour, metabolism and inflammation. These genes might reveal a molecular link between the two disorders. PMID- 17126560 TI - Growth hormone signalling: sprouting links between pathways, human genetics and therapeutic options. AB - Our molecular understanding of growth hormone-induced signal transduction has improved significantly over the past decades. At the same time, human population genetics and the analysis of genetically engineered animals have led to the discovery of genes that control specific aspects of the overall growth process. Although, currently, growth disorders are still diagnosed and treated on empirical bases, it might soon be possible to stratify patients predominantly by genetic defect, with treatment based on our molecular understanding of the role of the affected gene in the disease. PMID- 17126561 TI - Functional phosphoglucose isomerase from Mycobacterium tuberculosis H37Rv: rapid purification with high yield and purity. AB - Phosphoglucose isomerase (PGI) EC 5.3.1.9, is a housekeeping enzyme that catalyzes the reversible isomerization of d-glucopyranose-6-phosphate and d fructofuranose-6-phosphate. We have previously reported expression and multistep purification of recombinant PGI from Mycobacterium tuberculosis using conventional methods. We now describe an improved and simplified single step approach for purification of functionally active mycobacterial rPGI. The gene encoding PGI from M. tuberculosis H37Rv was cloned in bacterial expression vector pET22b(+). Expression of recombinant PGI with six-histidine-tag protein was observed both in the soluble fraction and inclusion bodies. Approximately 116mg of recombinant enzyme was purified to near homogeneity with approximately 80% yield from the soluble fraction of 1L culture at shake flask level using one step Ni-NTA affinity chromatography. The specific activity of the purified six histidine-tagged recombinant PGI (rPGI-His(6)) was approximately 800U/mg of protein. The apparent K(m) value of the active recombinant protein followed Michaelis-Menten kinetics and was 0.27+/-0.03mM. K(i) for the competitive inhibitor 6-phosphogluconate was 0.75mM. The enzyme had pH optima in the range of pH 7.6-9.0 and was stable up to 55 degrees C. rPGI-His(6) exhibited enzyme activity almost equal to that of enzyme without histidine tag. PMID- 17126562 TI - Functional expression and refolding of new alkaline esterase, EM2L8 from deep-sea sediment metagenome. AB - A metagenomics approach is an efficient method of isolating novel and useful genes from uncultured microorganisms in diverse environments. In this research, a gene encoding a new esterase (EM2L8) was cloned and characterized from the metagenomic DNA library of a deep-sea sediment. The gene consisted of 804bp encoding a polypeptide of 267 amino acids with a molecular mass of 28,952. The deduced amino acid sequence showed similarities with the BioH of Kurthia, the 3 oxoadipate enol-lactonase of Haloarcula and the acyltransferase of Thermoanaerobacter, which feature identities of 38%, 32%, and 33%, respectively. Residues essential for esterase activity, such as pentapeptide (GXSXG) and catalytic triad sequences, were uncovered. While the protein was overproduced mainly as inclusion body at 37 degrees C, it was mainly produced as a soluble active enzyme at 18 degrees C. A zymogram analysis revealed that purified EM2L8 taken from the soluble fraction could hydrolyze tributyrin substrate. Furthermore, the protein from the inclusion body fraction also showed strong activity on gel, thus indicating that the protein was refolded during SDS-gel electrophoresis and the ensuing incubation period. When the inclusion body was mixed with some anionic detergent solutions and diluted with a non-detergent buffer, the insoluble EM2L8 refolded rapidly and recovered its full esterase activity. Although EM2L8 had an optimum temperature of 50-55 degrees C, its activation energy in the range of 10-40 degrees C was 8.34kcal/mol, indicating that it is a cold-adapted enzyme. Moreover, it was found to have an optimum pH of 10-11, thus revealing that it is an alkaline enzyme. In this paper, the new esterase EM2L8 buried in a deep-sea sediment became known on the surface and was characterized biochemically. PMID- 17126563 TI - Refolding, purification, and activation of miniplasminogen and microplasminogen isolated from E. coli inclusion bodies. AB - Two des-kringle derivatives of human plasminogen, microplasminogen and miniplasminogen, have been expressed at high levels as inclusion bodies in Escherichia coli using a T7 expression system. In each case, the isolated inclusion bodies were refolded and purified. A final yield of approximately 10% of total refolded protein was observed in each case. Both refolded molecules were successfully activated to their functional forms, microplasmin and miniplasmin, by the plasminogen activator urokinase. The kinetic properties of the refolded microplasmin and miniplasmin were comparable to full length, native plasmin. PMID- 17126564 TI - Classification of continuous diffraction patterns: a numerical study. AB - The very intense and short pulses of future X-ray free electron lasers may allow the atomic resolution imaging of small, non-periodic objects. Preliminary estimates show that images obtained from single pulses do not contain statistically enough photons to allow successful reconstruction. Therefore multiple exposures of randomly oriented identical replicas have to be taken and the individual images have to be classified according to the object's orientation. The classification has been analytically treated by Huldt et al. [Huldt, G., Szoke, A., Hajdu, J., 2003. J. Struct. Biol. 144, 219.]. In this paper we extend the analytical results with numerical model calculations. This allows us to simulate realistic situations, which we will face in real experiments. We find significant deviations from the analytical expectations, even in the ideal case of spherical particles with random atomic distributions. We introduce a new norm for the individual scattering patterns and describe a criterion to select images belonging to similar orientation, which makes the classification more reliable in practice. We also discuss the effects of particle shape and size, partial orientational ordering, the measurement's resolution and the charge error caused by the Coulomb explosion. PMID- 17126565 TI - Conscious contributions to subliminal priming. AB - Choice reaction times to visual stimuli (targets) may be influenced by preceding subliminal stimuli (primes). Some authors reported a straight priming effect i.e., responses were faster when primes and targets called for the same response than when they called for different responses. Others found the reversed pattern of results. Eimer and Schlaghecken [Eimer, M. & Schlaghecken, F. (2002). Links between conscious awareness and response inhibition: evidence from masked priming. Psychonomic Bulletin &Review, 9, 514-520.] showed recently that straight priming occurs whenever a prime is not efficiently masked thereby the information provided by the prime is accessible for consciousness. In the present study, a hypothesis is tested that straight priming is due to mediation of consciousness. To test this hypothesis, prime validity was manipulated. We showed that even when no mask was used so that participants could fully and consciously perceive the prime and participants were informed that primes were mostly invalid, for the short prime-target ISI interval (100 ms) straight priming occurred. The priming was inverse when the ISI was 800 ms. This indicates that participants were able to use the information provided by the prime to prepare the response opposite to that cued by the prime but only if the time between the prime and the target was long enough. PMID- 17126566 TI - A study in the cognition of individuals' identity: solving the problem of singular cognition in object and agent tracking. AB - This article compares the ability to track individuals lacking mental states with the ability to track intentional agents. It explains why reference to individuals raises the problem of explaining how cognitive agents track unique individuals and in what sense reference is based on procedures of perceptual-motor and epistemic tracking. We suggest applying the notion of singular-files from theories in perception and semantics to the problem of tracking intentional agents. In order to elucidate the nature of agent-files, three views of the relation between object- and agent-tracking are distinguished: the Independence, Deflationary and Organism-Dependence Views. The correct view is argued to be the latter, which states that perceptual and epistemic tracking of a unique human organism requires tracking both its spatio-temporal object-properties and its agent-properties. PMID- 17126567 TI - A molecular phylogeny of Anopheles annulipes (Diptera: Culicidae) sensu lato: the most species-rich anopheline complex. AB - The Australasian Annulipes Complex is the most species-rich among Anopheles mosquitoes, with at least 15 sibling species suspected. Members of this complex are the most likely vectors of malaria in the past in southern Australia and are involved in the spread of myxomatosis among rabbits. In this, the first comprehensive molecular study of the Annulipes Complex, 23 ITS2 rDNA variants were detected from collections throughout Australia and Papua New Guinea, including diagnostic variants for the previously identified An. annulipes species A-G. Specimens of each ITS2 variant were sequenced for portions of the mitochondrial COI, COII and nuclear EF-1alpha genes. Partitioned Bayesian and Maximum Parsimony analyses confirmed the monophyly of the Annulipes Complex and revealed at least 17 clades that we designate species A-Q. These species belong to two major clades, one in the north and one mainly in the south, suggesting that climate was a driver of species radiation. We found that 65% (11) of the 17 sibling species recorded here had unique COI sequences, suggesting that DNA barcoding will be useful for diagnosing species within the Annulipes Complex. A comparison of the taxa revealed morphological characters that may be diagnostic for some species. Our results substantially increase the size of the subgenus Cellia in Australasia, and will assist species-level studies of the Annulipes Complex. PMID- 17126568 TI - Mitochondrial sequence diversity of the southernmost extant New World monkey, Alouatta caraya. AB - Variability in mitochondrial DNA sequences was analyzed in the howler monkey, Alouatta caraya, in order to delineate evolutionary relationships among populations in the most southerly distributed New World monkey. Based on new and previously published sequence data, fourteen cytochrome b haplotypes were observed among 33 howlers sampled in Argentina, Paraguay and Brazil, and grouped in two main haplogroups. In northeastern Argentina and southern Paraguay, new sequence data on 73 specimens sampled from six localities gave 34 control region haplotypes that also clustered in two main haplogroups. At this southern distribution, both mitochondrial markers revealed the presence of two sympatric and differentiated clades that we interpret to be the consequence of a secondary contact between previously allopatric populations. Given evidence for a demographic expansion at the beginning of the Holocene 15,500-7000 years ago (Fu's test, F(S)=-12.137; P<0.001), we suggest that atleast two populations of A. caraya have colonized the southernmost range since the Holocene employing forested corridors on the Parana and Paraguay Rivers. PMID- 17126569 TI - Death anxiety in patients with epilepsy. AB - PURPOSE: Whereas the relationship between epilepsy and anxiety has received much attention, less is known about the relationship between death anxiety and this disorder. The objective of this study was to assess death anxiety among epileptic patients who attended the outpatient neurology clinic at the Salmaniya Medical Complex, Kingdom of Bahrain. METHODS: Ninety-two patients (48 males and 44 females) completed a death anxiety scale. The scale items were adopted from already published surveys and adjusted to suit epilepsy patients. RESULTS: Results showed that the mean death anxiety score was moderate (2.75+/-1.35), with 26.09% of patients reporting high levels of death anxiety. Period of illness and educational level were significant predictors of death anxiety. Female patients, generalized type of epilepsy, the short duration of the illness and low level of education were associated with higher death anxiety scores. CONCLUSION: This study highlights the need for developing treatment strategies, counseling therapies and social support for people with epilepsy to decrease their death anxiety and improve their quality of life. PMID- 17126571 TI - Stressful stimuli modulate memory formation in Lymnaea stagnalis. AB - Stress has been shown to be a strong modulator of learning and memory in animals. We employ operant training of aerial respiratory behaviour in our model system, the pond snail Lymnaea stagnalis, to show that application of an acute consistent physical stressor enhances memory formation. A single 30 min operant conditioning training session, which normally results in intermediate-term memory (ITM) persisting 3h, results in long-term memory (LTM) persisting 24h if immediately preceded or followed by a stressor, for example a 30s exposure to 25 mM KCl. Other physical stressors (0.3% quinine-HCl or quick cooling and warming) similarly enhance memory formation. The memory is context specific and is not seen after the application of too much or too little stress. The memory can be extinguished by exposing snails to the hypoxic training environment and withholding reinforcing stimuli. The LTM that results from 30 min of training and stressor exposure is dependent on de novo protein synthesis and gene transcription in a single neuron, RPeD1. Because the soma of RPeD1 must be present for memory augmentation by the application of a stressor we are well placed for future investigations to directly determine the specific molecular alterations by which stress primes the formation of LTM. PMID- 17126570 TI - Synergistic effect of IGF-1 and OP-1 on matrix formation by normal and OA chondrocytes cultured in alginate beads. AB - OBJECTIVE: Growth factor therapy may be useful for stimulation of cartilage matrix synthesis and repair. Thus, the purpose of our study was to further understand the effect of combined insulin-like growth factor-1 (IGF-1) and osteogenic protein-1 (OP-1) treatment on the matrix synthesized by human adult normal and osteoarthritic (OA) chondrocytes. DESIGN: Chondrocytes were isolated post-mortem from articular cartilage from tali of normal human donors and femoral condyles of OA patients undergoing knee replacement surgery. Cells were cultured in alginate beads for 21 days in four experimental groups: (1) "mini-ITS" control; (2) 100 ng/ml IGF-1; (3) 100 ng/ml OP-1; (4) IGF-1+OP-1, each at 100 ng/ml. Beads were processed for histological (Safranin O and fast green), morphometrical and immunohistochemical (aggrecan, decorin, type I, II, VI, and X collagens, and fibronectin accumulation) analyses. RESULTS: Histology showed that IGF-1 alone did not induce substantial matrix production. OP-1 alone caused a considerable matrix formation, but the highest matrix accumulation by normal and OA chondrocytes was found when OP-1 and IGF-1 were added together. Morphometrical analysis indicated larger matrices produced by OA chondrocytes than by normal cells under the combined treatment. All tested matrix proteins were more abundant in the combination group. Type X collagen was detected only under the combined OP 1 and IGF-1 treatment and was present at very low levels. Type I collagen was found only in OA chondrocytes. CONCLUSIONS: The results obtained in the current study suggest that combined therapy with IGF-1 and OP-1 may have a greater potential in treating cartilage defects seen in OA than use of either growth factor alone. PMID- 17126572 TI - Where next after SPACE and EVA-3S: 'the good, the bad and the ugly!'. AB - SPACE and EVA-3S are the latest to publish outcomes in recently symptomatic patients who were randomised to carotid endarterectomy (CEA) or angioplasty and stenting (CAS). Contrary to expectations, both found that CAS was not 'as good as' CEA, while EVA-3S found CEA to be statistically superior. Not surprisingly, these trials have aroused considerable controversy and many in the pro-CAS lobby have simply dismissed them as being methodologically flawed and unrepresentative of contemporary CAS practice. However, to simply dismiss SPACE and EVA-3S as maverick trials is unacceptable. Unlike the landmark 'symptomatic' and 'asymptomatic' studies, the history of randomised trials comparing CAS and CEA has been characterised by repeated trial suspension (because of excess risk in the CAs cohort) and a systematic failure to achieve randomisation targets (thereby preventing any prospect of a statistically meaningful outcome) amid a milieu of corporate and individual conflicts of interest. SPACE and EVA-3S have certainly informed the dabate but they have not resolved it. Two trials are actively randomising recently symptomatic patients (ICSS in Europe and CREST in North America). Both require our support so that future guidelines are 'evidence based' rather than 'marker driven'. PMID- 17126573 TI - Genetic and antigenic typing of border disease virus isolates in sheep from the Iberian Peninsula. AB - A selection of 10 pestiviruses isolated from sheep from the Iberian Peninsula from 2001 to 2004 was characterised at the molecular level. The 5' untranslated region (5'-UTR) and N(pro)-coding gene were amplified by the reverse transcription-polymerase chain reaction (RT-PCR) and sequenced directly from purified products. All isolates were also typed antigenically with a panel of monoclonal antibodies (mAbs) raised against representative isolates of the four recognised pestivirus species. The genetic typing placed all the isolates in a new tentative type 4 of border disease virus (BDV), which was closely related to a pestivirus recently found in Pyrenean chamois (Rupicapra pyrenaica pyrenaica). Overall, the genotyping indicated a relatively wide diversity of the BDV type 4, which was best defined on the basis of N(pro) sequences. Antigenically, the isolates were recognised by two pan-pestivirus specific anti-NS3 mAbs, but only by some of the anti-glycoprotein specific mAbs raised against BDV, indicating partial antigenic overlap with other BDV isolates. PMID- 17126574 TI - Propofol reduces pulmonary edema by prevention of cytokine-mediated impairment of alveolar ion and fluid transport. PMID- 17126575 TI - Effects of salinity and temperature on the expression of enzymatic biomarkers in Eurytemora affinis (Calanoida, Copepoda). AB - In order to establish effective enzymatic biomarkers that could provide in situ early warning of contaminant exposure in estuarine ecosystems, the potential effects of the principal abiotic factors (temperature and salinity) were investigated on common biomarkers, the acetylcholinesterase (AChE) and the glutathione S-transferase (GST) in Eurytemora affinis. Short term salinity stress effects simulated during an experimental tide indicated that enzymatic activities of this species are characterized by maximum expression related to an optimal salinity range (between 5 and 15 psu). Moreover, longer time exposure to various salinity tanks confirmed the effects of this factor on both AChE and GST activities. Therefore, optimal AChE activity was measured at 10 psu, while optimal GST activity was measured at 5 psu. Furthermore, significant effects of temperature were also recorded, particularly for AChE expression (slight effects were measured on GST expression) with an optimal condition at 11 degrees C. These experiments indicated a more pronounced effect of salinity over temperature especially on the AChE expression and confirmed the need to standardize sampling procedures in relation with environmental parameters for biomonitoring studies based on enzymatic analyses. PMID- 17126576 TI - Ontogeny of energetics in leatherback (Dermochelys coriacea) and olive ridley (Lepidochelys olivacea) sea turtle hatchlings. AB - Changes in activity related oxygen consumption were measured in leatherback and olive ridley sea turtle hatchlings over their first month after emergence from the nest. Leatherbacks emerged with 75-90 KJ of energy in the residual yolk for growth and activity whereas olive ridleys emerged with 45 KJ. In leatherbacks (n=8), resting mass-specific oxygen consumption rates decreased by 53% over the first post-hatching month (0.34+0.03 mL O(2) h(-1) g(-1) to 0.16+0.01 mL O(2) h( 1) g(-1), respectively), while for ridleys (n=8) the fall was 35% (0.20+0.03 mL O(2) h(-1) g(-1) to 0.13+0.01 mL O(2) h(-1) g(-1), respectively). Olive ridley factorial aerobic scope doubled (1.93+0.30 to 3.97+0.51) over the first month but there was no significant increase in leatherback factorial aerobic scope (1.39+0.21 to 1.60+0.13). Leatherback hatchlings gained on average 20% initial body mass (7.68+1.66 g) over the first week, with 70 to 80% of this increase due to water accumulation. Olive ridleys gained 14% (1.83+0.16 g) in initial mass over the first week of age. We propose that the differences in aerobic scope and energy reserves are related to differences in early life ecological stratagems of these species. PMID- 17126577 TI - Daily changes in parameters of energy metabolism in brain of rainbow trout: dependence on feeding. AB - We assessed the daily patterns of parameters involved in energy metabolism in plasma and brain of rainbow trout. Where daily rhythms were found, we analyzed the potential influence of feeding. Immature rainbow trout were randomly distributed in 3 groups: fish fed for 7 days, fish fasted for 7 days, and fish fasted for 7 days and refed for 4 days. On sampling day, fish of fed and refed groups were fed at 11.00 h, and all fish were sampled from each treatment group using the following time schedule: 14.00, 18.00, 21.00, 00.00, 04.00, 07.00, 10.00 and 14.00 h. The results obtained from metabolic parameters assessed in plasma and brain can be grouped into three different categories, such as (i) those displaying no 24 h changes in fed fish such as plasma lactate, protein or acetoacetate levels, as well as brain amino acid and protein levels, and lowKm(glucose) hexokinase, and aspartate aminotransferase activities, (ii) those displaying 24 h changes that were apparently dependent on feeding since they disappeared in fasted fish such as the case of plasma cortisol, glucose and triglyceride levels, as well as brain glycogen, glucose, and lactate levels, and pyruvate kinase and hexokinase IV activities, and (iii) those parameters displaying 24 h changes apparently not dependent on feeding such as plasma amino acids, brain acetoacetate levels as well as several enzyme activities measured in brain such as glucose 6-phosphate dehydrogenase, alpha-glycerophosphate dehydrogenase, glutamate dehydrogenase, and lactate dehydrogenase-oxidase. In general, 24 h changes dependent on feeding indicate an increased use of glucose in brain several hours post-feeding whereas those changes not dependent on feeding were characterized by reduced levels/activity at the night period suggesting a metabolic depression in brain during darkness. PMID- 17126578 TI - Effects of exogenous cholecystokinin and gastrin on the secretion of trypsin and chymotrypsin from yellowtail (Seriola quinqueradiata) isolated pyloric caeca. AB - The humoral control of secretion of the proteolytic enzymes trypsin and chymotrypsin was studied in yellowtail (Seriola quinqueradiata). In vitro trials were performed to investigate the effects of cholecystokinin (CCK) and two commercially available gastrin peptides. Isolated preparations of pyloric caeca/pancreas release trypsin and chymotrypsin when incubated with cholecystokinin (CCK) at 10 microM and gastrin I (G1) at 50 microM after 15 min of incubation. On the other hand, G1 at 10 microM and gastrin-related peptide (G2) did not enhance trypsin and chymotrypsin secretion. The studies concerning the CCK effects at different incubation temperatures have shown that trypsin and chymotrypsin secretion at 25 degrees C was stimulated by CCK after 15 min, while at 10, 15 and 20 degrees C the stimulatory effects of CCK were observed only after 30 min of incubation. The CCK effects were increased at higher incubation temperatures and longer incubation periods. PMID- 17126579 TI - Phenotypic flexibility of digestive system in Atlantic cod (Gadus morhua). AB - This study examined the restoration of the digestive capacity of Atlantic cod (Gadus morhua Linnaeus) following a long period of food deprivation. Fifty cod (48 cm, 1 kg) were food-deprived for 68 days and then fed in excess with capelin (Mallotus villosus Muller) on alternate days. Ten fish were sampled after 0, 2, 6, 14 and 28 days and the mass of the pyloric caeca, intestine and carcass determined. Two metabolic enzymes (cytochrome c oxidase and citrate synthase) were assayed in white muscle, pyloric caeca and intestine, and trypsin activity was measured in the pyloric caeca. A delay of 14 days was required before body mass started to increase markedly, whereas most of the increase in mass of both the pyloric caeca and intestine relative to fish length occurred earlier in the experiment. By day 14, the activities of trypsin and citrate synthase in the pyloric caeca as well as citrate synthase in the intestine had reached maxima. The growth of the digestive tissues and restoration of their metabolic capacities thus occur early upon refeeding and are likely required for recovery growth to take place. The phenotypic flexibility of the cod digestive system is therefore remarkable: increases in trypsin activity and size of pyloric caeca resulted in a combined 29-fold increase in digestive capacity of the fish during the refeeding period. Our study suggests that Atlantic cod are able to cope with marked fluctuations in food availability in their environment by making a rapid adjustment of their digestive capacity as soon as food availability increases. PMID- 17126580 TI - The role of aquaporin 3 in teleost fish. AB - The aquaporin isoform, AQP3 has now been identified in a number of different teleost fish species, with additional DNA sequence information on AQP3 genes in further fish species available in genome databases. In zebrafish (Danio rerio), the AQP3 gene is present as two duplicate isoforms resulting from a teleostean fish genome-wide duplication. A further splicoform/isoform has also been identified in rainbow trout (Oncorhynchus mykiss). The identification of these AQP3 isoforms in other fish species is consequently explored. The role of AQP3 in physiological/osmoregulatory processes, in various teleost organs is then described. In teleost gill, AQP3 is expressed in 'chloride' cells, and in some species, in other epithelial cell types, where it may have a number of different functions including the prevention of dehydration. In eel esophagus, immunohistochemistry shows that AQP3 is expressed in surface epithelial cells in the anterior esophagus, but in mucus cells within the epithelium of the posterior esophagus. In eel intestine, AQP3 is found in macrophage-like cells and probably plays no part in osmoregulatory processes. In the rectum, as in the posterior esophagus AQP3 is expressed in mucus cells. In eel kidney, AQP3 is expressed in a subset of renal tubules, and localizes to the apical pole of tubule cells. There is no apparent change in the location or protein abundance of renal AQP3 following the acclimation of eels from freshwater to seawater. PMID- 17126581 TI - Stimulation of Mg2+-independent form of Ca2+-ATPase by a low molecular mass protein purified from goat testes cytosol. AB - A low molecular mass protein purified from goat (Capra hircus) testes cytosol following gel filtration and anion exchange chromatographic separation stimulates Mg(2+)-independent Ca(2+)-ATPase activity without any significant effect on Mg(2+)-dependent Ca(2+)-ATPase. Stimulation of the ATPase is due to an increase in the rate of dephosphorylation of the overall reaction step of the enzyme. Binding of the stimulator increases the affinity of Ca(2+)-ATPase for Ca(2+). An analysis of enzyme kinetics reveals a reversible type of binding of the stimulator to the ATPase and non-competitive type of stimulation with respect to the substrate. Stimulation seems due to binding of the protein at a single site following Michaelis-Menten model. The protein can also counter the effect of calcium antagonists exerted on the ATPase. The pI of the protein is 6.2 and its molecular mass has been determined to be 13, 961 by Q-TOF-MS. PMID- 17126582 TI - Sn-2-monoacylglycerol, not glycerol, is preferentially utilised for triacylglycerol and phosphatidylcholine biosynthesis in Atlantic salmon (Salmo salar L.) intestine. AB - Pathways of lipid resynthesis in the intestine of fish are relatively unknown. Various reports have suggested the existence of both sn-1,3-specific (pancreatic) and non-specific (bile salt-activated) lipase activity operating on dietary triacylglycerol (TAG) in the intestinal lumen of fish during digestion. Thus, sn 2-monoacylglycerol (2-MAG) and glycerol, respective hydrolytic products of each lipase, are absorbed and utilised for glycerolipid synthesis in enterocytes via two alternative routes: monoacylglycerol (MAG) and glycerol-3-phosphate (G3P) pathways. Despite different precursors, both pathways converge at the production of sn-1,2-diacylglycerol (1,2-DAG) where TAG or phosphatidylcholine (PC) synthesis can occur. To elucidate the relative activities of MAG and G3P pathways in Atlantic salmon enterocytes, intestinal segments were mounted in Ussing chambers where equimolar mixtures of sn-2-oleoyl-[1,2,3-(3)H]glycerol (2-MAG) and [(14)C(U)]glycerol, plus unlabelled 16:0 and 18:2n-6 as exogenous fatty acid sources, were delivered in bile salt-containing Ringer solution to the mucosa. The MAG pathway predominated, over the G3P pathway, synthesizing ca. 95% of total TAG and ca. 80% of total PC after a 3 h incubation period at 10 degrees C. Further, the 1,2-DAG branch point into TAG or PC was polarised towards TAG synthesis (6:1) via the MAG pathway but more evenly distributed between TAG and PC (1:1) via the G3P pathway. Effect of long-chain saturated, monounsaturated and polyunsaturated fatty acids on the synthesized TAG/PC ratio was assessed by individually exchanging 16:0, 18:1n-9 or 18:2n-6, for 16:0+18:2n-6, in mucosal solutions. TAG synthesis was influenced considerably more than PC synthesis, via either pathway, by exogenous fatty acids utilised. 18:1n-9 significantly stimulated TAG synthesis via the MAG pathway yielding a TAG/PC ratio of 12:1. Alternatively, 18:2n-6 stimulated TAG synthesis the most via the G3P pathway (TAG/PC=4:1). 16:0 significantly attenuated TAG synthesis via either pathway. Micellar fatty acid species also significantly affected intestinal active transport mechanisms as shown by decreasing transepithelial potential (TEP) and short-circuit current (SSC) with increasing fatty acid unsaturation. The epithelial integrity was, however, not compromised after 3 h of exposure to any of the fatty acids. The implications of these findings on dietary fatty acid composition and enterocytic lipid droplet accumulation are discussed. PMID- 17126583 TI - Induction of hemolin gene expression by bacterial cell wall components in eri silkworm, Samia cynthia ricini. AB - A cDNA clone encoding hemolin was isolated from fat body of immunized Samia cynthia ricini larvae based on subtractive suppression hybridization method. The cDNA encodes 413 amino acid residue open reading frame with an 18 residue predicted signal peptide. The expression of the gene was strongly induced in fat body and midgut by an injection of bacterial cells or peptidoglycans, but very weakly by lipopolysaccharide. The mRNA expression in the fat body was detected as early as 3 h post-injection, and reached the peak level at 12 h. PMID- 17126584 TI - Identification and partial characterisation of a chitinase from Nile tilapia, Oreochromis niloticus. AB - Measurement of chitinase activity in extracts from stomach, intestine, and serum of Nile tilapia with the artificial substrates 4-methylumbelliferil beta-D-N,N' diacetylchitobioside and 4-methylumbelliferil beta-D-N,N'N" triacetylchitotrioside (4MU[GlcNAc](2,3)) showed that an endochitinase was involved in the liberation of the fluorophore 4-methylumbelliferone (MU). Enzymes were isolated from tilapia serum by a combination of gel filtration, ion exchange, and reverse-phase chromatography. The molecular mass of the enzyme was estimated to be 75 kDa by SDS-PAGE, suggesting that the enzyme occurs as a monomer. The partially purified enzyme showed maximal activity at pH 7.0 when assayed with 4MU[GlcNAc](2) and lost its activity below pH 5.0 and above pH 8.0. The optimal pH of the purified enzyme toward the substrate 4MU[GlcNAc](3) was pH 9.0 and activity was lost below pH 8.0 and above pH 9.0. Our study has revealed the presence of a chitinolytic enzyme in the gastrointestinal tract and serum that may play a role in digestion and/or defense. PMID- 17126585 TI - The insulin-like growth factor (IGF)-system in active ulcerative colitis and Crohn's disease: relations to disease activity and corticosteroid treatment. AB - BACKGROUND: Metabolic bone disease (MBD) and muscle wasting (MW) are serious complications in adults suffering from inflammatory bowel disease (IBD). The inflammatory process and corticosteroid treatment may lead to changes in the IGF system associated with MBD and MW. AIM: To assess changes in the IGF-system and clinical and biochemical markers in ulcerative colitis (UC) and Crohn's disease (CD). METHODS: We studied 37 IBD patients with severe clinical exacerbation (20 with UC, 17 with CD) before and during high dose corticosteroid treatment and tapering (8-12 weeks). RESULTS: Total IGF-I was reduced in CD (36% p<0.01) and UC (41% p<0.001) before treatment and normalized completely. Free IGF-I baseline levels were unchanged compared to controls. In UC, free IGF-I levels increased significantly at week 1 and week 4 (p<0.01, respectively). In CD, no changes in free IGF-I levels were observed. IGFBP-2 baseline levels were increased by a factor 2.3 in UC and CD compared to controls (p<0.01 respectively) and normalized during treatment. IGFBP-3 was reduced by 38% (p<0.01) in CD and 32% (p<0.01) in UC with only partial normalization. Harvey-Bradshaw index, C - reactive protein and albumin normalized during treatment. CONCLUSIONS: Significant changes in total and free IGF-I and IGFBP-2 and IGFBP-3 were demonstrated in CD and UC patients in exacerbation with only partial normalization during high dose corticosteroid treatment and tapering without differences between UC and CD. These changes may be part of MBD and MW in active IBD. PMID- 17126586 TI - Expanded newborn screening identifies maternal primary carnitine deficiency. AB - Primary carnitine deficiency impairs fatty acid oxidation and can result in hypoglycemia, hepatic encephalopathy, cardiomyopathy and sudden death. We diagnosed primary carnitine deficiency in six unrelated women whose unaffected infants were identified with low free carnitine levels (C0) by newborn screening using tandem mass spectrometry. Given the lifetime risk of morbidity or sudden death, identification of adult patients with primary carnitine deficiency is an added benefit of expanded newborn screening programs. PMID- 17126587 TI - Neural agrin: a synaptic stabiliser. AB - Neural agrin is a heparan sulphate proteoglycan first defined by its ability to induce the clustering of acetylcholine receptors (AChRs) on cultured muscle cells. Neural agrin activates the transmembrane Muscle Specific Kinase (MuSK) on the postsynaptic muscle cell to stabilise the developing neuromuscular synapse. Three biological mechanisms for agrin/MuSK signalling are briefly discussed: selective transcription of synaptic genes such as MuSK itself, to reinforce developing postsynaptic clusters of AChRs; initiation of second messenger signalling pathways that can induce the formation of AChR clusters and retrograde signalling downstream of agrin/MuSK that may transform the growth cone of the motor axon into a stable differentiated nerve terminal, specialised for regulated exocytosis of neurotransmitter. Here we briefly review some key mechanisms through which neural agrin acts to foster the formation of mature neuromuscular synapses. PMID- 17126588 TI - Factors influencing the measurement of oxygen shortfall of the human cornea: sequencing of test conditions. AB - PURPOSE: The effects of sequencing of test conditions, in this case contact lens thicknesses, on the measurement of the oxygen shortfall of human corneas were studied. METHODS: Corneal oxygen uptake rates were measured with a Clark-type polarographic electrode on the central, unanesthetized right corneas of 14 human subjects. Measurements were made under the following conditions: (1) the normal open eye; (2) after 5 min of static (without blinking) wear of each of seven rigid gas permeable lenses of seven center thicknesses (0.18, 0.12, 0.16, 0.20, 0.24, 0.28, and 0.32 mm); (3) after 5 min of static wear of a polymethylmethacrylate (PMMA) contact lens. Lens thicknesses were randomly assigned numbers, which were sequenced in seven cycles. Two subjects were assigned to each sequencing cycle, and each subject participated in two identical sessions. RESULTS: The interaction of order x thickness was determined to be insignificant (F=0.99; p=0.5101). The effect of lens order was also insignificant (F=0.76; p=0.6239), indicating that the order of lens placement did not affect the measured corneal oxygen shortfall. Not surprisingly, the analysis indicated a significant effect of lens thickness on corneal oxygen shortfall (F=3.94; p=0.0032). CONCLUSIONS: The sequencing of lenses of various thicknesses on the cornea does not affect the measurement of corneal oxygen shortfall. PMID- 17126589 TI - iNOs expression is stimulated by the major surface protein (rWSP) from Wolbachia bacterial endosymbiont of Dirofilaria immitis following subcutaneous injection in mice. AB - The bacterial endosymbiont Wolbachia of several species of filarial nematodes plays an important role in the inflammatory pathology of filariasis. Nitric oxide (NO) production has also been implicated in the immune response during filarial infections. Here we present data indicating that a recombinant Wolbachia surface protein (rWSP) induces iNOs mRNA expression and NO production, as well as IFN gamma and a Th1-type antibody response, in inoculated BALB/c mice. This effect is not observed when mice are inoculated with a recombinant heat shock protein from Wolbachia (GroEL). PMID- 17126590 TI - Synthesis and spectroscopic studies on chromium(III) complex containing mixed valence chrysenesemiquinone-chrysenecatecholate ligands and 2,2'-bipyridine coligand. AB - The synthesis and spectroscopic properties of Cr(bpy)(chrySQ)(chryCat), a complex containing chromium(III) metal ion and chrysenequinone ligand in its partially reduced (chrySQ) and fully reduced (chryCat) forms, are described. The complex has been prepared by two different routes from Cr(CO)6 and Cr(chrySQ)3. Variable temperature magnetic susceptibility measurements indicated a strong antiferromagnetic coupling between Cr(III) (S=3/2) and chrysenesemiquinone radical (S=1/2), giving a magnetic coupling constant J=-342 cm(-1). Ligand-based redox couples were observed in the electrochemical studies that consist of quasi reversible chrySQ/chryCat and bpy/bpy*- reductions and chryCat/chrySQ oxidation at negative potentials and irreversible chrySQ/chryBQ oxidation at positive potential. However, the metal was inert in the studied potential range. The electronic spectra of the complex revealed interesting properties. In addition to interaligand pi-pi* and n-pi* transitions, other bands corresponding to Cr(t(2g)) ->chrySQ(pi*) and Cr(t(2g))-->bpy(pi*) metal-to-ligand charge-transfer MLCT transitions were observed. The infrared spectral analysis was informative in assigning the vibrations due to SQ and Cat ligands. Also, it was a useful tool in confirming the coordination of bpy ligand to chromium metal ion. PMID- 17126591 TI - Investigation of the electronic vibrational structure of furan by REMPI. AB - Resonance-enhanced multiphoton ionization (REMPI) has been applied to detect the hot bands as well as the cold bands of the 1a2-->-->3dxy(1A1) Rydberg transition of furan (jet-cooled, mass-analyzed). Based on the unambiguous assignment of the hot bands, a complete vibronic analysis is given for the cold bands of this transition (up to 4600 cm(-1) above the origin). This analysis can be used for the interpretation of the vibrational structure in the X 2A2 photoelectron band. The energy ordering of the five 3d Rydberg states is 1A2 approximately 2A260 years. Data from the National Health and Nutrition Examination Survey 1999-2002 were used in this study. STATISTICAL ANALYSIS: Group means were estimated for the age group, sex, and race/ethnicity subgroups. The probability that any of these group means were equal to one another was tested using statistical software. RESULTS: The data showed marked differences in beverage consumption depending on age, sex, and race/ethnicity. In general, males consumed more beverages than did females. Specifically, white and Mexican-American persons of all ages consumed more milk than did African-American persons. On average, African-American males and females of all ages consumed significantly more fruit drinks/ades than did other race/ethnicity groups. In contrast, white persons consumed more carbonated soft drinks than did other race/ethnicity groups. CONCLUSION: Average beverage consumption varied depending on age, sex, and race/ethnicity. Knowledge of differences in beverage consumption patterns is important for food and nutrition professionals and nutrition policymakers. Better understanding of the many factors that influence beverage consumption levels is needed. PMID- 17126631 TI - Food preparation by young adults is associated with better diet quality. AB - OBJECTIVE: To describe food-preparation behaviors, cooking skills, resources for preparing food, and associations with diet quality among young adults. DESIGN: Cross-sectional analyses were performed in a sample of young adults who responded to the second wave of a population-based longitudinal study. Measures pertaining to food preparation were self-reported and dietary intake was assessed by a food frequency questionnaire, both by a mailed survey. SUBJECTS/SETTING: Males (n = 764) and females (n = 946) ages 18 to 23 years. STATISTICAL ANALYSES PERFORMED: Cross-tabulations and chi2 tests were used to examine associations between food preparation, skills/resources for preparing foods, and characteristics of young adults. Mixed regression models were used to generate expected probabilities of meeting the Healthy People 2010 dietary objectives according to reported behaviors and skills/resources. RESULTS: Food-preparation behaviors were not performed by the majority of young adults even weekly. Sex (male), race (African American), and living situation (campus housing) were significantly related to less frequent food preparation. Lower perceived adequacy of skills and resources for food preparation was related to reported race (African American or Hispanic) and student status (part-time or not in school). The most common barrier to food preparation was lack of time, reported by 36% of young adults. Young adults who reported frequent food preparation reported less frequent fast-food use and were more likely to meet dietary objectives for fat (P < 0.001), calcium (P < 0.001), fruit (P < 0.001), vegetable (P < 0.001), and whole-grain (P = 0.003) consumption. CONCLUSIONS: To improve dietary intake, interventions among young adults should teach skills for preparing quick and healthful meals. PMID- 17126632 TI - Predictors for research involvement among registered dietitians. AB - OBJECTIVE: The primary aim of this study was to measure registered dietitians' (RDs') research involvement (by creating a research score) and to determine whether their perceptions, attitudes, and knowledge of evidence-based practice and key antecedent factors (eg, sociodemographic characteristics, education and training, professional experiences, and employment setting) predicted their research involvement. DESIGN: This cross-sectional, descriptive study used the Dietitian Research Involvement Survey and followed the Tailored Design Method. SUBJECTS/SETTING: This study surveyed 258 randomly selected RDs from seven dietetic practice groups of the American Dietetic Association. STATISTICAL ANALYSES PERFORMED: Descriptive statistics, bivariate relationships, and multiple linear regression analyses were conducted to test whether perceptions, attitudes, and knowledge of evidence-based practice score and antecedent factors predicted the research score of dietitians. RESULTS: Perceptions, attitudes, and knowledge of evidence-based practice score (r = 0.59, P < 0.0005), level of education (r = 0.53, P < 0.0005), taking a research course (r = 0.40, P < 0.0005), last time read research (r = 0.35, P < 0.0005), frequency of professional reading (r = 0.32, P < 0.0005), primary area of practice (r = 0.14, P = 0.02), association memberships (r = 0.14, P = 0.02), and dietetic practice group affiliation (r = 0.14, P = 0.02) were significantly correlated with research score. Using multivariate linear regression, the perceptions, attitudes, and knowledge of evidence-based practice score (beta = 0.48, P < 0.0005) and level of education (beta = 0.39, P < 0.0005) were identified as the strongest predictors of research score. CONCLUSIONS: Involvement in research by RDs is largely determined by their perceptions, attitudes, and knowledge of evidence-based practice and their level of education. Additional education and training related to research methodology and design, and evidence-based practice, is essential for greater participation in research activities by RDs. PMID- 17126633 TI - Vitamin supplement intake is related to dietary intake and physical activity: The Child and Adolescent Trial for Cardiovascular Health (CATCH). AB - OBJECTIVE: To explore the relationship of multiple-vitamin supplement use with selected food groups, physical activity, lifestyle behaviors, and weight status. SUBJECTS AND METHODS: Two thousand seven hundred sixty-one adolescents in the 12th grade who participated in the fourth Child and Adolescent Trial for Cardiovascular Health study had height and weight measured and completed health behavior survey and food frequency questionnaires. Logistic regression models were used to determine the likelihood of supplement use with health and activity behaviors and dietary intake. RESULTS: Prevalence of multiple-vitamin supplement use among adolescents was 25% and varied by sex and race/ethnicity. Supplement users had higher mean daily intakes of most food groups, but lower intakes of total fat and saturated fat than nonusers. Higher food index scores were positively associated with the likelihood of using multiple-vitamin supplements. Supplement users were more likely to be physically active, participate in team and organized sports, and less likely to be overweight and to watch more than an hour of television per day. CONCLUSIONS: Adolescents who use multiple vitamin supplements have more healthful dietary and lifestyle behaviors than nonusers. Further study on supplement use by adolescents, including other types of supplements used and reasons for use, is warranted. PMID- 17126634 TI - Motivational interviewing for pediatric obesity: Conceptual issues and evidence review. AB - Counseling by health care professionals represents a potentially important intervention for the prevention and treatment of pediatric obesity. One promising approach to weight-control counseling in pediatric practice is motivational interviewing. This article explores conceptual issues related to the application of motivational interviewing for the prevention and treatment of pediatric obesity. Given the paucity of studies on motivational interviewing and pediatric obesity, we examine what is known about the application of motivational interviewing to modify diet, physical activity, and other behaviors in children and adolescents. We begin with a brief overview of motivational interviewing, describe some nuances of applying this approach to pediatric overweight, and conclude with research and clinical recommendations. PMID- 17126635 TI - Factors influencing implementation of the Coordinated Approach to Child Health (CATCH) Eat Smart School Nutrition Program in Texas. AB - The purpose of this research was to evaluate factors influencing the implementation of the Coordinated Approach to Child Health (CATCH) Eat Smart School Nutrition Program in Texas using data from the CATCH dissemination study. A mail survey was sent to school foodservice personnel (N = 213) who attended a CATCH training from August 2000 through January 2002. A response rate of 40% (n = 85) was achieved. The mean score for the percentage of CATCH Eat Smart guidelines implemented was 80.44. Multivariate linear regression analysis revealed that, after adjusting for age and number of years employed in school foodservice, the following factors were significantly associated with the percentage of CATCH Eat Smart guidelines implemented: utility of CATCH and CATCH Eat Smart in meeting requirements for Coordinated School Health Programs (P = 0.006), school foodservice personnel's satisfaction with food made using the CATCH Eat Smart guidelines (P = 0.008), utility of CATCH in facilitating interschool communication about children's health (P = 0.019), and perceived student satisfaction with food made using the CATCH Eat Smart guidelines (P = 0.046). These results suggest that dissemination approaches for Coordinated School Health Programs should focus on ways to enhance program satisfaction, be consistent with legislated mandates, and increase interschool staff communication to increase program implementation by school foodservice personnel. PMID- 17126636 TI - Availability of weight-loss supplements: Results of an audit of retail outlets in a southeastern city. AB - The sale of nonprescription weight-loss products accounts for millions of dollars spent by Americans trying to lose weight, yet there is little evidence for effectiveness and there are multiple safety concerns. The purpose of this study was to determine what products, and ingredients within products, were available at retail outlets in a metropolitan area. A purposive sampling strategy identified 73 retail outlets. An audit form was used to collect information from product labels. The audit identified 402 products containing 4,053 separate ingredients. The mean number of ingredients per product was 9.9+/-8.96 (range = 1 to 96). A database search was conducted regarding evidence for effectiveness, safety precautions, and side effects for the 10 ingredients that appeared most often across products. Modest evidence of effectiveness exists for green tea (Camellia sinensis), chromium picolinate, and ma huang (Ephedra major). For the remaining seven (ginger root [Zingiber officinale], guarana [Paullinia cupana], hydroxycitric acid [Garcinia cambogia], white willow [Salix alba], Siberian ginseng [Eleutherococcus senticosus], cayenne [Capsicum annuum], and bitter orange/zhi shi [Citrus aurantium]), inadequate or negative evidence exists. Although precautions and contraindications were found for all 10 ingredients, the strongest concerns in the literature appear for ma huang, bitter orange, and guarana. Our audit revealed numerous weight-loss products available to consumers, yet there is little evidence to support the effectiveness of the top 10 ingredients identified and many potential adverse reactions; therefore, food and nutrition professionals should discuss dietary supplement use with their clients. PMID- 17126637 TI - Existence and predictors of soft drink advertisements in Pennsylvania high schools. AB - The objective of this study was to describe the extent and locations of soft drink advertisements on high school campuses in Pennsylvania and identify factors related to extent of these advertisements. Surveys were distributed to 271 school foodservice directors in a random sample of high schools in Pennsylvania. These high schools were selected to be representative of the entire population of high schools in Pennsylvania based on chosen demographic characteristics. A three phase survey strategy was used, involving distribution of a postcard reminder 1 to 2 weeks after the initial survey distribution, and mailing of a second survey to nonrespondents 1 to 2 weeks after mailing of the postcard. Two hundred twenty eight school foodservice directors (84%) returned surveys. Linear multiple regression analyses were done using SPSS (version 11.5.1, 2002, SPSS Inc, Chicago, IL). Approximately two thirds (66.5%) of respondents indicated soft drink advertisements exist in at least one location in their school, with the most prevalent locations being on vending machines (62%) and school grounds, such as playing fields (27%). Slightly more than 10% of respondents indicated soft drink advertisements displayed in the cafeteria. Extent of soft drink advertisement locations was positively related to existence of a pouring-rights contract, subscription to Channel One, and receipt of incentives from soft drink bottlers based on sales, but negatively related to average daily participation in school lunch. These findings suggest that commercialization and sales incentives might interact to contribute to school environments that are not "nutrition friendly." Schools' efforts to establish wellness policies as mandated by the Child Nutrition and WIC Reauthorization Act of 2004 provide ideal opportunities to examine school environments for advertising that might conflict with the healthful environments they are aiming to establish, and perhaps to develop policies to address these practices. PMID- 17126638 TI - Cranberry does not affect prothrombin time in male subjects on warfarin. AB - There have been case reports suggesting that cranberry beverages may interact with warfarin. To date, no research study has been conducted to examine the potential interaction of cranberry and warfarin. The current study is a randomized, placebo-controlled, double-blind, crossover study to investigate the effect of cranberry juice on prothrombin time as assessed by the international normalized ratio (INR). Seven subjects with atrial fibrillation on a stable dose of warfarin for 3 months were randomized to consume 250 mL of cranberry juice for 7 days, then placebo for 7 days, or vice versa. The washout period was 7 days. The prothrombin time/INR was measured at baseline, and on days 2, 4, 7, 10, 14, 16, 18, 21, and 24. Data were analyzed by the Student t test for paired values. The baseline INR was 2.28+/-0.54 for the cranberry group and 2.13+/-0.50 for the placebo group. For all test points, the INR did not change significantly from baseline. At day 7 on cranberry juice, the INR was 2.23+/-0.53 for cranberry first group and 2.16+/-0.40 for placebo first group. The mean differences between the cranberry and placebo groups were not statistically significant. Our results suggest no significant interaction between the daily consumption of 250 mL cranberry juice and warfarin. When counseling patients on dietary changes necessary during warfarin treatment, it does not seem necessary to eliminate daily cranberry juice consumption at amounts of 250 mL, but the INR should be followed up closely. PMID- 17126639 TI - Clinical registered dietitians, employers, and educators are interested in advanced practice education and professional doctorate degrees in clinical nutrition. AB - A subset of registered dietitians (RDs) is known to practice at an advanced level, but a clear educational pathway supporting advanced medical nutrition therapy practice has not been identified. Thus, an electronic survey was designed to investigate interest of clinical RDs, employers, and educators in advanced practice competencies and professional doctorate degree programs in clinical nutrition. Usable responses were obtained from 440 of 978 (45%) RDs, 61 of 107 (57%) employers, and 76 of 114 (67%) educators. Mean interest (5 = very interested, 1 = very uninterested) in obtaining advanced practice education was highest among RDs (3.93+/-1.01) and was significantly different (P < 0.01) from employers (3.74 +/-1.28) and educators (2.76+/-1.33). Interest in completing a professional doctorate in clinical nutrition was significantly (P < 0.01) different among RDs (3.05+/-1.28), employers (3.18+/-1.30), and educators (2.3+/ 1.34). Employers' mean interest score for hiring RDs with a professional doctorate in clinical nutrition was 4.02+/-0.93. A subset of clinical RDs appears to be interested in obtaining advanced practice competency and enrolling in professional doctorate degrees in clinical nutrition. Clinical nutrition managers in academic medical centers may be interested in hiring advanced practice clinical RDs with professional doctorate degrees. Opportunities exist for educators to develop advanced practice educational experiences and professional doctorate degree programs. PMID- 17126640 TI - Dietary quality and associated factors among adults living in the state of Sao Paulo, Brazil. AB - This study sought to analyze dietary quality and associated factors among adults living in regions of the State of Sao Paulo, Brazil. It was a cross-sectional population-based study of a sample of 3,454 adults ages 20 years and over who were included in the Household Health Survey. Dietary intake was measured by means of the 24-hour recall method, and dietary quality was assessed by means of the Healthy Eating Index (HEI), adapted to local realities. Probabilistic samples were obtained via multistage cluster samples from four regions in the State of Sao Paulo. Linear regression analyses were performed to evaluate the relationships between the HEI and the demographic, socioeconomic, and lifestyle variables. Among the individuals assessed, 5% had a good diet, 74% a diet that needed some degree of improvement, and 21% a poor diet. The means for HEI components were lowest for vegetables, fruits, and dairy products. The highest HEI scores were obtained by individuals who were nonsmokers, practiced physical activity, were retired, lived in houses or apartments, and had adequate living conditions surrounding them. In the multiple regression analysis, the variables of numbers of consumer durable goods, schooling of the head of the family, energy intake, and age had a positive association with the HEI. However, the association was inverse for the variables of smoking and body mass index. Higher dietary quality is associated with higher income, higher schooling level, better nutritional status, and being a nonsmoker. Knowledge of these factors is important for implementing programs for preventive nutrition or intervention. PMID- 17126641 TI - Low-carbohydrate diets: Assessing the science and knowledge gaps, summary of an ILSI North America Workshop. PMID- 17126642 TI - Relation between plaque progression and low-density lipoprotein cholesterol during aging as assessed with serial long-term (> or =12 months) follow-up intravascular ultrasound of the left main coronary artery. AB - Because of the clinical benefit of lipid lowering in older patients, we hypothesized that the relation between low-density lipoprotein (LDL) cholesterol serum levels and coronary plaque progression may persist throughout aging. We analyzed serial intravascular ultrasound (IVUS) data of 60 left main stems (18 +/ 9 months apart) and evaluated the relation between LDL cholesterol levels and coronary plaque progression at different ages. The population (n = 60) was divided into 3 groups according to age: tertile 1 (n = 20) was a mean age of 48 +/- 6 years (median 51, range 33 to 55), tertile 2 (n = 20) was a mean age of 58 +/- 2 years (median 59, range 55 to 61), and tertile 3 (n = 20) was a mean age of 66 +/- 6 years (median 65, range 61 to 83). Between groups, there was no significant difference in non-age-related demographics, clinical data, lipid profiles, or medications (e.g., statins). There was a positive linear relation between LDL cholesterol and annual changes in plaque plus media area in all age tertiles, which was statistically significant in tertiles 2 and 3 (r = 0.56, p <0.01; r = 0.50, p <0.02) and showed a strong trend in tertile 1 (r = 0.41, p = 0.07). The estimated LDL cholesterol thresholds, which, as determined by regression analysis, would correspond to no plaque progression, were 74, 60, and 78 mg/dl, respectively, in tertiles 1, 2, and 3. In conclusion, serial IVUS data in left main coronary arteries suggest that the relation between LDL cholesterol serum levels and plaque progression persists during aging. PMID- 17126643 TI - Long-term prognostic role of flow-mediated dilatation of the brachial artery after acute coronary syndromes without ST elevation. AB - Coronary endothelial vasodilator dysfunction is associated with increased cardiac events; the close relation between coronary vasomotor dysfunction and brachial artery vasoreactivity has been previously described. This study assessed the prognostic value of noninvasively assessed brachial artery vasoreactivity in survivors of acute coronary syndromes without ST-segment elevation. We examined 98 men (63.1 +/- 10.8 years) who were referred to our hospital for acute coronary syndromes without ST-segment elevation. Brachial artery endothelium-dependent flow-mediated dilation (FMD) and endothelium-independent nitrate-mediated dilation were examined in all patients using high-resolution echocardiographic Doppler ultrasound within 24 hours of admission. Plasma malondialdehyde, a marker of oxidative stress, and left ventricular ejection fraction were also assessed. Twenty-seven patients underwent coronary revascularization. Patients were followed for 24.8 +/- 5.9 months. Cardiovascular death, myocardial infarction, stroke, and unstable angina were designated as cardiovascular events (CEs). Twenty CEs were recorded. Kaplan-Meyer analysis showed that patients with FMD <1.9% (tertile 1 of FMD values) were more likely to have CEs than those with FMD >1.9% (log rank 5.29, p = 0.021). Multivariate Cox regression analysis showed that FMD <1.9% predicted CEs with an adjusted hazard ratio of 3.035 (95% confidence interval 1.148 to 8.023, p = 0.025) after adjustment for age, risk factors, troponin T, ejection fraction, revascularization procedures, number of diseased vessels, and medication. In conclusion, endothelium-dependent dilation of the brachial artery is a strong independent predictor of adverse outcome in survivors of acute coronary syndromes without ST-segment elevation. PMID- 17126644 TI - Prevalence of metabolic syndrome after acute coronary syndrome and its prognostic significance. AB - Metabolic syndrome (MS) consists of a cluster of metabolic and hemodynamic disorders that promote the development of atherosclerosis and increase cardiovascular morbidity/mortality. We evaluated the prevalence and characteristics of MS after acute coronary syndrome (ACS) and the effect of intensive risk factor management on the morbidity/mortality associated with MS in a therapeutic cohort; 480 consecutive patients were summoned 3 months after an ACS for cardiovascular evaluation and management. Follow-up was carried out 16 months later. At 3 months after ACS, prevalence of MS was 20.8%, as assessed by criteria of the National Cholesterol Education Program Adult Treatment Panel III and 27.7% according to the definition of the International Diabetes Federation. The most common metabolic disorders were abdominal obesity, hypertriglyceridemia, and fasting hyperglycemia. Characteristics of the initial ACS showed no significant difference between the MS and non-MS groups. Atherosclerotic extent was greater in the MS group according to Adult Treatment Panel III. At follow-up, the MS and non-MS groups achieved optimal low-density lipoprotein cholesterol and blood pressure levels. During follow-up, there was an increase in total mortality in the MS group compared with the non-MS group (5.2% vs 1.4%, p <0.01) as assessed by International Diabetes Federation criteria; however, no difference in minor or major cardiovascular events was found between the 2 groups. In conclusion, MS was highly prevalent after an ACS, notably in young patients, and was not associated with a specific ACS presentation. PMID- 17126645 TI - Effect of clopidogrel plus aspirin on ST segments in patients with ST-elevation acute myocardial infarction. AB - Spontaneous ST-segment resolution (STR) after acute ST-elevation myocardial infarction is associated with favorable outcomes. Effect on STR of newer, more powerful antiplatelet and antithrombotic agents is unclear. The aim of this study was to identify independent clinical and angiographic predictors of STR in patients with ST-elevation acute myocardial infarction before percutaneous coronary intervention. We studied 206 patients admitted with ST-elevation acute myocardial infarction, of whom 37 (18%) had STR. There were 12 deaths (5.8%) that occurred in the group without spontaneous STR. Patients with spontaneous STR were younger (55 vs 61 years old, p = 0.02), had shorter duration of symptoms (117 vs 212 minutes, p <0.0001), had preserved ejection fraction (55% vs 40%, p <0.0001), had shorter hospital stays, and had lower in-hospital arrhythmias or death. Independent predictors of STR, identified by stepwise logistic regression analysis, were early clopidogrel administration (odds ratio [OR] 2.10, 95% confidence interval [CI] 1.00 to 4.40, p = 0.045), single-vessel disease (OR 2.85, 95% CI 1.22 to 6.70, p = 0.02), chest pain duration (OR 0.98, 95% CI 0.98 to 0.99, p <0.0001), collaterals (OR 4.3, 95% CI 1.7 to 10.8, p = 0.002), circumflex as a culprit vessel (OR 4.74, 95% CI 1.5 to 14.95, p = 0.008), and coronary thrombus noted on angiography (OR 5.76, 95% CI 1.63 to 20.4, p = 0.006). In conclusion, early clopidogrel administration is associated with, and likely causal for, STR. Patients with thrombus, collateral flow, and circumflex culprit vessel are more likely to have STR. In addition, our study confirms previous findings that patients with STR have preserved left ventricular function and better hospital outcomes compared with those without STR. PMID- 17126646 TI - Prognostic value of intraventricular dyssynchrony according to clinical stage of left ventricular impairment. AB - Intraventricular dyssynchrony has prognostic implications in patients who have severe functional limitation and decreased ejection fraction. Patients with less advanced cardiac disease often exhibit intraventricular dyssynchrony, but there is little available information about its prognostic relevance in such patients. We investigated the prognostic effect of intraventricular dyssynchrony on outcome in 318 patients with known or suspected coronary artery disease who were classified according to the presence or absence of left ventricular dysfunction and heart failure symptoms. Mortality was considered the primary end point over a median follow-up of 56 months, and a Cox proportional hazards model was used for survival analysis. Despite a low prevalence (8%) of left bundle branch block, there was a high prevalence of intraventricular dyssynchrony even in patients without symptomatic heart failure. The magnitude of intraventricular dyssynchrony correlated poorly with QRS duration (r = 0.25), end-systolic volume index (r = 0.27), and number of scar segments (r = 0.25). There were 58 deaths during follow up. Ventricular volume, ischemic burden, and magnitude of intraventricular dyssynchrony predicted outcome, but magnitude of intraventricular dyssynchrony was an independent predictor of survival only in patients with asymptomatic left ventricular dysfunction. In conclusion, patients with known or suspected coronary artery disease have a high prevalence of intraventricular dyssynchrony. Although ventricular volume, ischemic burden, and intraventricular dyssynchrony are potentially important prognostic markers, the relative importance of intraventricular dyssynchrony changes with the clinical setting and may be greatest in patients with preclinical disease. PMID- 17126647 TI - Effect of gender on prognosis following percutaneous coronary intervention for stable angina pectoris and acute coronary syndromes. AB - Women with non-ST-elevation acute coronary syndromes (NSTACSs) may have better outcomes than men, but the effect of NSTACSs in women undergoing percutaneous coronary intervention (PCI) has not been examined. We performed a prospective, multicenter, cohort study of consecutive patients who underwent PCI for NSTACS and stable angina during 3 National Heart, Lung, and Blood Institute Dynamic Registry recruitment waves (1997 to 2002) to examine the effect of female gender on adverse clinical events after PCI or stable angina for NSTACS. The primary end point was the combined rate of death, myocardial infarction, or rehospitalization for cardiac causes at 1 year. Compared with men with NSTACS (n = 2,124), women (n = 1,338) were older and more often had hypertension, diabetes mellitus, and history of heart failure (p <0.001 for all), whereas multivessel disease was less frequent (p <0.01). Procedural success and in-hospital adverse event rates were similar. Women with NSTACS had the highest 1-year rate of death/myocardial infarction/cardiac rehospitalization compared with women with stable angina pectoris (n = 462) or men (n = 995; women with NSTACS 37.6%, men with NSTACS 29.8%, women with stable angina 29.4%, men with stable angina 27.7%, p <0.001). The higher rate remained after adjustment for differences in baseline characteristics (adjusted hazard ratio 1.37, 95% confidence interval 1.20 to 1.56). Among women, NSTACS conferred a significantly higher risk for adverse events compared with stable angina (adjusted hazard ratio 1.41, p = 0.001), whereas the risk of adverse events was not different in men (adjusted hazard ratio 1.05, p = 0.5). In conclusion, women undergoing PCI for NSTACS have a higher risk of major adverse cardiac events than men or women undergoing PCI for stable angina. PMID- 17126648 TI - Comparison with conventional therapies of repeated sirolimus-eluting stent implantation for the treatment of drug-eluting coronary stent restenosis. AB - This study compared the safety and efficacy of repeat percutaneous coronary intervention (PCI) using sirolimus-eluting stents (SESs) with conventional therapies for restenosis after drug-eluting stent placement. Fifty-five consecutive patients with 58 restenotic lesions (31 treated with SESs and 27 treated with paclitaxel-eluting stents) underwent PCI using SESs (33 lesions) or conventional therapies comprising cutting balloon angioplasty alone (11 lesions) or intracoronary brachytherapy (14 lesions). Baseline characteristics were similar for the 2 groups, except for greater edge involvement (75.8% vs 36.0%, p = 0.002) and less stent expansion (0.74 +/- 0.17 vs 0.95 +/- 0.21, p = 0.006) in the SES group than in the conventional group. The SES group achieved a greater postprocedural luminal gain than the conventional group (1.98 +/- 0.50 vs 1.22 +/ 0.48 mm, p <0.001). Follow-up angiography showed that late luminal loss (0.27 +/ 0.56 vs 0.76 +/- 0.84 mm, p = 0.021) and recurrent angiographic restenosis rate (3.6% vs 35.0%, p = 0.006) were lower in the SES group than in the conventional group. The repeated target lesion revascularization-free survival rates at 1 year were 96.7 +/- 3.2% for the SES group and 91.7 +/- 5.6% for the conventional group (p = 0.399). In conclusion, use of SESs was associated with a lower recurrent restenosis rate compared with conventional therapies. PMID- 17126649 TI - Safety and efficacy of the 2.25-mm sirolimus-eluting Bx Velocity stent in the treatment of patients with de novo native coronary artery lesions: the SIRIUS 2.25 trial. AB - Smaller reference vessel diameter is a recognized determinant of in-stent restenosis. The SIRIUS 2.25 trial was a prospective, nonrandomized study including 100 patients (mean age 63.4 years; 64% men, 40% with diabetes mellitus) assessing the safety and efficacy of the 2.25-mm sirolimus-eluting Bx Velocity stent in patients with de novo native coronary lesions. Using propensity score matching for gender, diabetes mellitus, left anterior descending artery target vessel, lesion length, and reference vessel diameter, the outcomes were compared with historical control groups (angioplasty and Palmaz-Schatz stent arms from the STRESS/BENESTENT I/II trials and the Bx Velocity bare metal stent arm from the RAVEL and SIRIUS trials having a reference vessel diameter <3 mm). Use of the 2.25-mm sirolimus-eluting Bx Velocity stent was associated with a high rate of procedural success (97%) and a low rate of in-hospital major adverse cardiac events (2%). The primary end point, 6-month in-lesion binary angiographic restenosis, occurred less frequently in patients treated with the 2.25-mm sirolimus-eluting Bx Velocity stent than in each of 3 historical controls (16.9% vs 30.6%, p = 0.12; 36.5%, p <0.001; 45.9%, p <0.001, respectively). This translated into lower rates of 6-month target lesion revascularization in the 2.25-mm sirolimus-eluting Bx Velocity stent group (4.0% vs 15.0% in each of 3 control groups, p = 0.01 to <0.001). By multivariate analysis, in-lesion binary restenosis was predicted by multiple implanted stents (odds ratio 10.4, p = 0.002). Four of 13 patients who developed restenosis (30.8%) had a diffuse pattern of restenosis. In the long lesion tertile (mean lesion length 19.5 mm), the in-lesion binary restenosis rate was 27.6%. In conclusion, use of the 2.25-mm sirolimus-eluting Bx Velocity stent was safe and provided favorable 6-month clinical outcomes. Use of multiple stents (in longer lesions) was an independent predictor of in-lesion restenosis. PMID- 17126650 TI - Safety and efficacy of short-term celecoxib before elective percutaneous coronary intervention for stable angina pectoris. AB - Fifty patients with stable angina pectoris entered a randomized, double-blind study and were assigned to receive celecoxib (200 mg 2 times daily) or placebo 7 days before percutaneous coronary intervention (PCI). Results showed that detection of markers of myocardial injury above the upper normal limit was significantly lower in the celecoxib than in the placebo group: 12% versus 35% for creatine kinase-MB (CK-MB; p = 0.001), 20% versus 48% for troponin I (p = 0.0004), and 22% versus 51% for myoglobin (p = 0.0005). Myocardial infarction by CK-MB determination was less commonly seen after PCI in the celecoxib than in the placebo group (5% vs 18%, p = 0.025). Postprocedural peak levels of CK-MB (2.9 +/ 18 vs 7.5 +/- 18 ng/ml, p = 0.0002) were also significantly lower in the celecoxib than in the placebo group. No significant side effect was reported by the 2 groups of patients. In conclusion, pretreatment with celecoxib 200 mg 2 times daily for 7 days significantly decreased procedural myocardial injury in elective PCI. These findings indicate that the antiphlogistic action of cyclo oxygenase-2 inhibition may provide a friendly protection to ischemic cardiomyocytes. PMID- 17126651 TI - Comparison of the efficacy of direct coronary stenting with sirolimus-eluting stents versus stenting with predilation by intravascular ultrasound imaging (from the DIRECT trial). AB - A direct coronary stenting technique using drug-eluting stents may decrease drug eluting stent efficacy due to possible damage to the surface coating of the stent. The DIRECT is a multicenter, prospective, nonrandomized trial designed to evaluate the direct stenting strategy for the sirolimus-eluting Bx-Velocity stent compared with the historical control (SIRIUS trial, stenting with predilation). Volumetric and cross-sectional intravascular ultrasound analyses at 8-month follow-up were performed in 115 patients (DIRECT n= 64, control n = 51). Patient and lesion characteristics were comparable between groups. The DIRECT group achieved an equivalent uniform expansion index, defined as minimum stent area/maximum stent area x 100, compared with the control group (65.9 +/- 11.7 vs 63.1 +/- 12.7, p = NS). At 8-month follow-up, vessel, stent, lumen, and neointimal volume index (volume in cubic millimeters/length in millimeters) and percent neointimal volume were similar between the DIRECT and control groups (vessel volume index 13.9 +/- 4.40 vs 15.0 +/- 3.83; stent volume index 6.83 +/- 2.02 vs 6.94 +/- 2.04; lumen volume index 6.71 +/- 2.04 vs 6.81 +/- 2.07; neointimal volume index 0.14 +/- 0.24 vs 0.16 +/- 0.23; percent neointimal volume 3.73 +/- 6.97 vs 3.14 +/- 5.32, p = NS for all). In addition, in-stent neointimal hyperplasia distribution was significantly smaller near the distal stent edge (0.22 vs 0.098 mm(3)/mm, p = 0.01 for an average neointimal volume index within 3 mm from the distal stent edge). In conclusion, direct coronary stenting with the sirolimus-eluting Bx-Velocity stent is equally effective in terms of uniform stent expansion and long-term quantitative intravascular ultrasound results compared with conventional stenting using predilation. This strategy appears to be associated with less neointimal hyperplasia near the distal stent edge. PMID- 17126652 TI - Competing impact of excess weight versus cardiorespiratory fitness on cardiovascular risk. AB - Obesity is a risk factor for cardiovascular disease, whereas high cardiorespiratory fitness (CRF) is cardioprotective. This study evaluated the competing effect of weight and fitness on biomarkers of cardiovascular risk in a nationally representative sample of 2,112 adults (20 to 49 years of age; body mass index [BMI] > or =18.5 kg/m(2)) without previously diagnosed cardiovascular disease from the National Health and Nutrition Examination Survey 1999 to 2002. CRF levels were assigned using age- and gender-specific reference points of estimated maximal oxygen consumption calculated from submaximal graded exercise treadmill testing. CRF was also categorized by sample-specific tertiles of maximal oxygen consumption. Weight was categorized using BMI. Fasting insulin level >12.2 mU/L, C-reactive protein level > or =3.0 mg/L, and total cholesterol/high-density lipoprotein ratio (TC/HDL) >5 characterized increased cardiovascular risk. CRF and BMI were independently associated with increased fasting insulin and C-reactive protein (p <0.05). When patients with low, moderate, and high CRF were further stratified as normal, overweight, or obese, weight remained significantly associated with increased fasting insulin, C reactive protein, and TC/HDL (p <0.001), but CRF did not. Logistic regressions evaluating increased fasting insulin, C-reactive protein, and TC/HDL demonstrated no significant differences in overweight/obese patients by CRF level after adjustment for other factors. Significant differences were present between normal weight and overweight or obese patients regardless of fitness level. Analyses using tertiles of CRF yielded similar results. In conclusion, patients who are "fat but fit" require weight-loss interventions to improve their cardiovascular risk profiles. Future interventions should emphasize weight control, even for those with high CRF. PMID- 17126653 TI - Multifactor cardiovascular disease risk reduction in medically underserved, high risk patients. AB - Few data exist on the effectiveness of cardiovascular disease (CVD) risk reduction programs in patients with limited access to health care. The objective of this project was to evaluate a disease management approach to multifactor CVD risk reduction in patients with limited or no health insurance and low family income. Patients (n = 148) were recruited from not-for-profit or free clinics and hospitals and randomized to usual care or usual care plus team case management. Mean age was 59.3 years, 57% were women, 50% had less than a high school education, 57% were Hispanic, and 64% had no health insurance. All had > or =1 increased risk factor for CVD, and 24.5% had documented coronary artery disease. Follow-up measurements were obtained at 6 and 12 months. Primary outcomes were low-density lipoprotein cholesterol and systolic blood pressure. The disease management program was supervised by a physician, delivered by nurses and dietitians, and included comprehensive lifestyle changes and medications. Data were collected on 91% of patients at 12 months. Disease management produced clinically important decreases in selected risk factors compared with usual care, including systolic blood pressure (p <0.01) and low-density lipoprotein cholesterol (p <0.03). More patients with disease management than those with usual care moved from "high" and "very-high" risk to lower risk categories for selected risk factors. In conclusion, the disease management program had excellent retention and lower CVD risk factors and demonstrated the potential of such approaches for decreasing long-term disease burden in selected medically underserved populations. PMID- 17126654 TI - Predictors of clinical outcomes in patients given carvedilol for heart failure. AB - Risk factors for outcomes in heart failure (HF) were derived from populations in clinical trials, at hospital discharge, or in localized geographic or socioeconomic strata before the widespread use of beta blockers. This study observed 4,280 patients in a community-based HF registry for 1 year after completing carvedilol titration. Independent risk factors for death, hospitalization for HF, or hospitalization for cardiovascular reasons other than HF were first identified by age-, gender-, and race-adjusted analyses, then by multivariate analysis adjusted simultaneously for all factors. Over this period, 7% of patients died, 11% were hospitalized for HF, 12% were hospitalized for other cardiovascular reasons, and 27% had > or =1 of these events. The most significant outcome predictors were New York Heart Association class III or IV, history of hospitalization for HF or other cardiovascular reasons, and angina pectoris, all associated with increased odds of having an adverse outcome (all p < or =0.001). The left ventricular ejection fraction was not a significant outcome predictor by multivariate analysis. The odds ratio for an adverse outcome was significantly reduced for patients with hypertensive or idiopathic causes of HF and for those whose physicians had graduated from medical school > or =24 years earlier compared with <14 years earlier (all p <0.005). In conclusion, easily obtained historical information predicts clinical outcomes in patients with HF in the year after initiating carvedilol. In this unselected community population, these historical factors were better predictors of risk than the left ventricular ejection fraction. PMID- 17126655 TI - Comparison of 60-day mortality in hospitalized heart failure patients with versus without hypothermia. AB - The use of aggressive treatments and the modification of current treatment in patients with heart failure (HF) relies heavily on the assessment of disease severity using prognostic markers. However, many such markers are unavailable in routine clinical practice, and others have little prognostic value. This study tested the hypothesis that low body temperature could predict short-term survival after discharge in patients hospitalized for HF. Data from the Acute and Chronic Therapeutic Impact of a Vasopressin Antagonist in Congestive Heart Failure (ACTIV in CHF) trial, which randomized 319 patients hospitalized for HF to receive placebo or tolvaptan, were retrospectively analyzed. Hypothermia was defined a priori as an oral body temperature <35.8 degrees C at randomization. Cox regression was used to analyze survival within a 60-day follow-up period. Hypothermia was observed in 32 patients (10%). Mortality rates at 60 days after discharge were 6.3% (20 of 319) overall, 9.4% (3 of 32) in hypothermic patients, and 5.9% (17 of 287) in nonhypothermic patients. Hypothermia was a strong multivariate predictor of mortality; hypothermic patients were 3.9 times more likely to die within 60 days than nonhypothermic patients (95% confidence interval 1.002 to 15.16, p = 0.0497) after adjustment for treatment group, age, and other confounders. Hypothermia was associated with such indicators of low cardiac output as an elevated blood urea nitrogen/creatinine ratio, narrow pulse pressure, and a reduced ejection fraction. In conclusion, hypothermia appears to be a strong predictor of mortality in patients with HF. PMID- 17126656 TI - Effects of levosimendan on right ventricular function in patients with advanced heart failure. AB - Right ventricular (RV) dysfunction frequently complicates advanced left ventricular heart failure and contributes to an unfavorable prognosis. Levosimendan is a novel inodilator that beneficially affects hemodynamics and left ventricular systolic and diastolic function in patients with advanced heart failure. However, its effects on RV function have not yet been properly assessed in these patients. In this randomized trial, the impact of levosimendan or placebo on various echocardiographic parameters of RV systolic and diastolic function was investigated in 54 patients with advanced heart failure due to left ventricular systolic dysfunction. Tissue Doppler imaging maximal systolic tricuspid annular velocity (S wave) increased significantly only in the levosimendan group (8.2 +/- 3.2 vs 9.0 +/- 3.0 cm/s, p <0.03). Tissue Doppler imaging RV early diastolic velocity (E wave) and the ratio of early to late diastolic velocities (E/A) also increased significantly after levosimendan administration (p <0.01 and p <0.05, respectively). Systolic pulmonary arterial pressure decreased significantly (54 +/- 11 vs 43 +/- 11 mm Hg, p <0.01) in the levosimendan-treated patients. Levosimendan beneficially modulated neurohormonal and inflammatory status by decreasing B-type natriuretic peptide levels (p <0.05) and by altering the ratio of interleukin-6 to interleukin-10 in favor of the latter (p <0.05). In conclusion, levosimendan could offer further therapeutic advantages in patients with advanced heart failure by improving systolic and diastolic RV function. PMID- 17126657 TI - Relation of patient age to outcome of percutaneous mitral valvuloplasty. AB - The aim of this study was to evaluate the impact of age on immediate- and long term outcomes of percutaneous mitral balloon valvuloplasty (PMV). PMV is the first-line treatment for patients with symptomatic mitral stenosis. However, long term results in large series of patients from Europe and the United States have been found less favorable than those from Asia and South America involving younger patients. Six hundred ten patients who underwent 626 PMV procedures were prospectively followed for 6.1 +/- 4.10 years using clinical and echocardiographic evaluation. Patients were divided in quartiles according to age: < or =41 years (n = 163), 42 to 53 years (n = 163), 54 to 63 years (n = 142), and >63 years (n = 158). The success of PMV was defined as valve area > or =1.5 cm(2) without severe regurgitation; restenosis was defined as a loss > or =50% of initial gain, with a valve area of <1.5 cm(2). PMV success was significantly more prevalent in younger patients: 95.7% in group 1, 91.4% in group 2, 86.4% in group 3, and 83.4% in group 4 (p = 0.002). No significant differences in complications were found among all age groups, including death, cardiac tamponade, emergency mitral replacement, and any embolic events (p = NS). Event-free survival was greater in younger patients (p <0.0001), but on multivariate analysis, age was not an independent predictor of events (p = NS). Restenosis occurred in 27.9% of patients, throughout all groups (p = NS). In conclusion, PMV may be safely and effectively performed in younger and older patients. Although event-free survival was greater in younger groups, multivariate analysis did not find that age was an independent predictor of events. PMID- 17126658 TI - Effectiveness of same day percutaneous coronary intervention followed by minimally invasive aortic valve replacement for aortic stenosis and moderate coronary disease ("hybrid approach"). AB - In 2005, the investigators described a "hybrid" cardiovascular interventional strategy combining percutaneous coronary intervention (PCI) for coronary artery disease (CAD) followed by valve surgery for patients with urgent complex CAD and valve disease to reduce morbidity and mortality. This hybrid approach has been extended prospectively in elderly, high-risk patients with aortic stenosis scheduled for elective minimally invasive aortic valve replacement (MI-AVR) who, on preoperative coronary angiography, were found to have moderate CAD amenable to PCI. In this prospective, observational series, 18 patients (mean age 76 years) underwent elective hybrid MI-AVR with PCI from May 2003 to February 2006. Five patients had undergone previous coronary artery bypass grafting. Patients underwent coronary angiography the day of (n = 12) or evening before (n = 6) MI AVR, and after identifying moderately severe CAD, all 18 underwent the implantation of drug-eluting stents to the affected coronary arteries, followed by MI-AVR. Although all patients received standard doses of antiplatelet medications, including acetylsalicylic acid (325 mg before PCI and 325 mg/day thereafter) and clopidogrel (300 mg after PCI, 75 mg/day thereafter for 90 days for the Cypher stent), there were no reoperations for bleeding; only 8 of 18 patients required postoperative blood transfusions. One patient died postoperatively from a colonic perforation, and there were no late mortalities after a mean follow-up of 19 months. In conclusion, this hybrid strategy has low morbidity and mortality and may be a new therapeutic option for older, high-risk patients with combined CAD and aortic valve disease. PMID- 17126659 TI - Usefulness of N-terminal pro-B-type natriuretic peptide to predict clinical course in patients with hypertrophic cardiomyopathy. AB - This study was designed to assess the value of plasma N-terminal pro-B-type natriuretic peptide (NT-pro-BNP) levels in predicting clinical courses in 80 patients with hypertrophic cardiomyopathy. Patients were followed for a mean of 571 +/- 294 days for clinical end points, defined as a composite of cardiovascular death and hospitalization for worsening heart failure symptoms. NT pro-BNP plasma level (hazard ratio 1.6, 95% confidence interval 1.10 to 2.55, p = 0.015) and New York Heart Association (NYHA) functional class (hazard ratio 3.7, 95% confidence interval 1.17 to 12.09, p = 0.025) were the independent variables associated with increased risk for experiencing clinical end points. NT-pro-BNP plasma level > or =1,500 pg/ml detected patients with clinical end points with sensitivity, specificity, and accuracy of 83%, 81%, and 81%, respectively. The hypertrophic cardiomyopathy population was stratified into low-, medium-, and high-risk subgroups using 2 simple variables, NYHA functional class and NT-pro BNP. Patients in lower NYHA classes and with NT-pro-BNP levels <1,500 pg/ml were significantly free of clinical end points. In conclusion, for patients with hypertrophic cardiomyopathy, plasma levels of NT-pro-BNP seem a reliable parameter to identify those at risk for clinical deterioration at long-term follow-up in conjunction with symptomatic status. PMID- 17126660 TI - Determinants for clinical diagnosis of hypertrophic cardiomyopathy. AB - Although hypertrophic cardiomyopathy (HC) occurs in 1 of 500 adults, most cardiology practices treat relatively few patients with HC, suggesting that many affected patients evade clinical recognition. Determining the clinical circumstances under which HC is identified will provide clues to its under recognition. Clinical triggers leading to diagnostic echocardiograms were analyzed in 711 consecutive patients with HC. In most (384 [54%]), HC was initially suspected only after the onset of cardiac symptoms or acute cardiac events. In a substantial minority (327 [46%]), HC was recognized while patients were asymptomatic, including 225 (32%) by routine medical evaluations, in 27 of whom (4%) HC was recognized during preparticipation examinations for competitive sports or other activities. Women, older patients (age > or =50 years), and those with outflow obstruction at rest (gradient > or =30 mm Hg) were more likely suspected to have HC by virtue of cardiac symptoms or events (p <0.0001). Conversely, patients with extreme hypertrophy (wall thickness > or =30 mm) and those at high risk for sudden death were more often asymptomatic and identified by routine or family screenings (p <0.0001 and p = 0.004, respectively). Patients who subsequently died of heart failure or experienced embolic stroke were more often identified by virtue of symptoms or acute events (p = 0.03). In conclusion, although most patients with HC were recognized clinically only after overt disease manifestations, a substantial minority were diagnosed by routine examinations while asymptomatic, including an important subset of patients with HC recognized solely because of findings on sports preparticipation screening. These data underscore the need for heightened awareness and clinical suspicion of HC to increase the number of diagnosed patients, including many who may be at high risk for sudden death. PMID- 17126661 TI - Electrocardiographic findings in 888 patients > or =90 years of age. AB - The oldest old are the fastest growing component of many national populations. No large-scale analysis of electrocardiographic findings in this group exists to date. This study cataloged the frequencies of specific electrocardiographic (ECG) findings in the target population and compared the frequencies of these findings by gender and race. The electrocardiograms of 888 subjects aged > or =90 years presenting for any reason were retrospectively analyzed using standard criteria for 128 separate ECG findings. Left ventricular enlargement (28%), first-degree atrioventricular block (16%), and atrial fibrillation (15%) were the most common abnormalities found in the population as a whole. In contrast to previous studies, which demonstrated a predominance of left-axis deviation, 91% of the electrocardiograms analyzed in our study demonstrated a mean frontal QRS axis ranging from -30 degrees to +90 degrees . Sinus rhythm was observed in 79% of the population. More Caucasians than African-Americans had atrial fibrillation (19% vs 12%, p = 0.004). Women had more normal ECG results than men (6% vs 2%, p = 0.02), and more women demonstrated complete right bundle branch block (13% vs 7%, p = 0.002). In conclusion, few patients aged > or =90 years who have their electrocardiograms recorded at hospitals have completely normal ECG results. The frequencies of some ECG findings in the target population vary by gender and race. PMID- 17126662 TI - Impact of glomerular filtration rate on minor troponin T elevations for risk assessment in patients undergoing operation for abdominal aortic aneurysm or lower extremity arterial obstruction. AB - Debate surrounds the impact of renal function on the prognostic value of minor troponin T release in vascular surgery patients. The objective of this study was to assess the long-term prognostic value of minor degrees of troponin T release in patients who undergo major vascular surgery, especially those with concomitant renal dysfunction. Survivors of major noncardiac vascular surgery (n = 558) were preoperatively screened for cardiac risk factors and renal function. Serial troponin T was measured on days 1, 3, and 7 after surgery, using a threshold of 0.03 ng/ml. All-cause mortality and major adverse cardiac events (MACEs) were noted during follow-up (mean 3.5 +/- 2.0 years). Minor (0.03 to 0.09 ng/ml) and major (> or =0.1 ng/ml) release of troponin T was observed in 5% and 8%, respectively. During follow-up, 21% of the patients died and 15% experienced MACEs. After adjustment for the estimated glomerular filtration rate, patients with minor and major troponin T release were at comparable increased risk for late mortality (hazard ratio [HR] 3.43, 95% confidence interval [CI] 1.79 to 6.58, and HR 3.72, 95% CI 2.37 to 5.85, respectively), and MACEs (HR 5.47, 95% CI 2.60 to 11.48, and HR 6.32, 95% CI 3.82 to 10.48, respectively) compared with patients with troponin T release <0.03 ng/ml. Tests for heterogeneity revealed that minor and major troponin T release have prognostic value across the entire spectrum of renal function. In conclusion, marginal elevations of troponin T strongly predict late mortality and MACEs after major vascular surgery, irrespective of renal function. A currently underestimated high-risk subgroup of patients may be identified using a lower troponin T threshold. PMID- 17126663 TI - Relation of weights of intraaneurysmal thrombi to maximal right-to-left diameters of abdominal aortic aneurysms. AB - We determined the weight of operatively excised thrombi within abdominal aortic aneurysms (AAAs) in 42 patients aged 52 to 92 years (mean 73 +/- 9). The thrombi in the 32 men ranged in weight from 12 to 586 g (mean 162 +/- 135) and in the 10 women, from 12 to 351 g (mean 94 +/- 102). The maximal right-to-left diameter of the AAA by computed tomography immediately preoperatively in the 32 men ranged from 5.0 to 11.0 cm (mean 7.6 +/- 1.7), and in the 10 women from 4.0 to 10.5 cm (mean 6.7 +/- 1.9). The relation of the weight of the intraaneurysmal thrombus to the maximal right-to-left diameter of the AAA preoperatively was highly significant in both men (r = 0.72, p <0.001) and women (r = 0.88, p <0.001). In conclusion, the intraaneurysmal thrombi consisted virtually entirely of fibrin, indicating no evidence of organization. PMID- 17126664 TI - Evidence of left ventricular contractile asynchrony by echocardiographic phase imaging in patients with type 2 diabetes mellitus and without clinically evident heart disease. AB - Left ventricular electromechanical asynchrony has been shown to predict cardiac events in patients with heart failure. This study investigated whether left ventricular asynchrony is present in patients with type 2 diabetes mellitus (DM) with no clinically evident heart disease and normal QRS durations. Asynchrony was evaluated in 24 patients with DM, 15 nondiabetic control subjects, and 20 patients with left bundle branch block (LBBB) due to cardiomyopathy serving as positive controls by conventional tissue Doppler imaging and by a novel method, echocardiographic phase imaging. Asynchrony was significantly higher in patients with DM than in controls and significantly lower than in patients with LBBB. This was shown by tissue Doppler imaging: the SD of time to peak myocardial velocity was 13 +/- 10 ms in controls, compared with 30 +/- 19 ms in patients with DM (p <0.01) and 68 +/- 28 ms in those with LBBB (p <0.001). Similar data were obtained using echocardiographic phase imaging: the SD of phase degrees was 25 degrees +/- 8 degrees in controls, compared with 44 degrees +/- 21 degrees in patients with DM (p = 0.02) and 76 degrees +/- 25 degrees in those with LBBB (p <0.001). Tissue Doppler imaging correlated with echocardiographic phase imaging (r = 0.79, p <0.0001) but was more time consuming (15.5 +/- 4.5 vs 4.5 +/- 2.2 min/patient, p <0.05) and showed higher intraobserver variability (5.6% vs 3.2%, p <0.05). In conclusion, this is the first study showing increased left ventricular asynchrony in patients with DM and no clinical evidence of heart disease. PMID- 17126665 TI - Usefulness of automated quantitation of regional left ventricular wall motion by a novel method of two-dimensional echocardiographic tracking. AB - A novel 2-dimensional (2-D) echocardiographic tracking method enables the automated tracking of displacement between 2 points of interest in echocardiographic images. The purpose of this study was to evaluate whether this 2-D tracking method could be used to measure left ventricular (LV) wall thickness and percent systolic wall thickening (%WT) in parasternal short-axis views, in comparison with conventional manual measurement, and to determine whether this system can be used to quantitatively assess regional LV wall motion in echocardiography. In 24 subjects (12 with LV wall motion abnormalities), 6 segments in the short-axis images were assessed by this method. Two sample points at the endocardium and epicardium were tracked automatically during 1 cardiac cycle. Then, LV wall thickness and %WT were calculated. In 50 subjects (32 with LV wall motion abnormalities), average %WT as an average of all %WT in every degree of angle in each of the 6 segments was measured by this method, and the results were compared with the visual assessment of LV wall motion scores. There was excellent agreement between the 2-D tracking and manual methods for LV wall thickness (r = 0.99) and %WT (r = 0.97). The mean differences in LV wall thickness and %WT were 0.1 +/- 0.4 mm and 0 +/- 5.4%, respectively. Average %WT was significantly decreased in the regions of hypokinetic or akinetic wall motion compared with those of normal motion (18 +/- 4% and 4 +/- 4% vs 39 +/- 10%, p <0.001). In conclusion, this 2-D tracking method can be used for the noninvasive, automated quantitation of LV wall motion in 2-D echocardiography. PMID- 17126666 TI - Effects of acute administration of caffeine on vascular function. AB - Caffeine is the most widely used pharmacologic substance in the world. It is found in common nonessential grocery items (e.g., coffee, tea, cocoa, and chocolate). The effects of caffeine on cardiovascular diseases, including hypertension, remain controversial, and there is little information on its direct effect on vascular function. The purpose of this study was to determine the effect of caffeine on endothelial function in humans. This study was a double blind, randomized placebo and active drug study. Forearm blood flow (FBF) responses to acetylcholine (ACh), an endothelium-dependent vasodilator, and to sodium nitroprusside, an endothelium-independent vasodilator, were evaluated in healthy young men before and after the oral administration of caffeine 300 mg (n = 10) or placebo (n = 10). FBF was measured by using a strain-gauge plethysmograph. Caffeine significantly increased systolic and diastolic blood pressures by 6.0 +/- 6.0 and 2.6 +/- 3.1 mm Hg (p <0.05), respectively, but did not alter heart rate or baseline FBF. Caffeine augmented the FBF responses to ACh from 21.2 +/- 7.1 to 26.6 +/- 8.1 ml/min/100 ml tissue (p <0.05), whereas sodium nitroprusside-stimulated vasodilation was not altered by caffeine administration. The intra-arterial infusion of N(G)-monomethyl-L-arginine, a nitric oxide synthase inhibitor, abolished the caffeine-induced augmentation of FBF response to ACh. In the placebo group, the ACh- and sodium nitroprusside-stimulated vasodilation was similar before and after the follow-up period. In conclusion, these findings suggest that the acute administration of caffeine augments endothelium-dependent vasodilation in healthy young men through an increase in nitric oxide production. PMID- 17126667 TI - Emerging strategies for increasing high-density lipoprotein. AB - High-density lipoprotein cholesterol is a potent and independent epidemiologic risk factor and is a proved antiatherosclerotic agent in animal models of atherosclerosis, acting through the principal mechanisms of accelerating cholesterol efflux and inhibiting oxidation and inflammation. Lifestyle modification increases serum levels by 5% to 15%, whereas niacin, the drug most widely used to increase high-density lipoprotein cholesterol, increases it by 25% to 35% at the highest doses. This review examines the potent methods of increasing high-density lipoprotein and/or enhancing reverse cholesterol transport, including cholesterol ester transfer protein inhibitors, apolipoprotein A-I Milano, D4F, the dual peroxisome proliferator-activated receptor agonists, and rimonabant, that are now in clinical trials. In conclusion, these new agents, used alone or in combination with existing therapies, carry the potential to markedly reduce the incidence of new coronary disease and cardiac events in this decade. PMID- 17126668 TI - Atherosclerosis: its cause and its prevention. PMID- 17126669 TI - Prognostic role of body mass index after acute myocardial infarction. PMID- 17126670 TI - Putting emphasis on the first part of a biochemical cascade. PMID- 17126671 TI - Prediction of outcome in patients undergoing surgery for tricuspid regurgitation. PMID- 17126672 TI - Impact of enzyme replacement therapy on cardiac morphology and function and late enhancement in Fabry's cardiomyopathy. PMID- 17126673 TI - Cardiomyopathy and myopathy in left ventricular noncompaction. PMID- 17126675 TI - Beyond lipid lowering: What have we learned about the benefits of statins from the acute coronary syndromes trials? AB - The long-term benefits for prevention of primary and secondary cardiovascular events achieved by lowering serum cholesterol levels with statins are undisputed. However, results of early clinical trials and preclinical studies suggested the possibility that benefits of treatment with the 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) may not be entirely due to their effects on low-density lipoprotein (LDL) cholesterol. Since then, large-scale clinical trials of statin treatment for acute coronary syndromes (ACS) have demonstrated early benefits that are associated with aggressive initiation of statin therapy. Support for pleiotropic effects of statins in these trials has been provided not only by these acute effects but also by apparent differences in efficacy between various statin regimens that seem unrelated to their effects on serum cholesterol levels. Analyses of biomarkers of inflammation, oxidation, and thrombosis in samples from patients in these studies have also suggested possible mechanisms underlying the benefits of statins in ACS. This article reviews some of the key statin trials in ACS and assesses the evidence for benefits of these drugs independent of their effects on LDL cholesterol. PMID- 17126676 TI - Inflammatory biomarkers in stable atherosclerosis. AB - Inflammation is a key mechanism in atherosclerotic progression and destabilization that precedes clinical events such as myocardial infarction. The inflammatory biomarkers provide a window into many of these inflammatory processes. In research studies, average levels of these biomarkers in groups of subjects are often related to the risk of clinical events, and modification of risk factors can change the plasma concentrations of many biomarkers, reflecting suppression of inflammation. More evidence exists for C-reactive protein (CRP) than for other inflammatory biomarkers, and the discussion of the clinical value of plasma levels of these markers is focused mainly on CRP. Although the inflammatory biomarkers are useful research tools, their value as a clinical instrument for assessment of cardiovascular risk and/or treatment efficacy is still widely debated. Each biomarker has passionate supporters who advocate these applications, but, at this stage, no inflammatory biomarker has universal support for clinical use and their potential for fulfilling this role requires further study. PMID- 17126677 TI - Plaque stabilization: Can we turn theory into evidence? AB - Coronary events often result from thrombi that form because of physical disruption of the atherosclerotic plaque. The dynamic nature of the plaque offers the opportunity to intervene to modify plaque biology with lifestyle changes and, if needed, pharmacologic measures. Inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A reductase (statins) reduce levels of serum cholesterol and decrease the incidence of coronary events, but some of the benefits of statins may not depend on their effects on circulating lipids. Indeed, increasing evidence suggests that statins may also enhance plaque stability. Such evidence includes results of preclinical studies with experimental atherosclerosis as well as imaging data and analyses of proinflammatory and prothrombotic mediators in clinical trials. Currently, however, no studies have demonstrated conclusively the mechanisms underlying the unexpected magnitude and rapidity of statin benefits. This article reviews the evolution of the concept of plaque stabilization and reexamines the evidence for the role of statins in that process. PMID- 17126678 TI - Molecular basis of differences among statins and a comparison with antioxidant vitamins. AB - Although all statin drugs lower levels of circulating low-density lipoprotein cholesterol by inhibiting 3-hydroxy-3-methylglutaryl coenzyme A, molecular differences among statins affect their metabolism, solubility, and intramembrane localization, which in turn is likely to influence their efficacy and safety. In addition, these properties may have a differential impact on the pleiotropic effects of statins, including their ability to improve endothelial function and to affect proliferation and apoptosis in vascular tissues. Many pleiotropic effects of statins appear to be due to inhibition of small guanosine triphosphate binding proteins and/or restoration of nitric oxide bioavailability, and atorvastatin metabolites may also directly protect vascular tissues from oxidative damage. The possibility of cardiovascular benefits from antioxidant effects of atorvastatin metabolites contrasts with the lack of benefits seen in most studies of antioxidant vitamins. This article reviews some of the differences in pleiotropic effects of statins and assesses the contribution of their solubility and membrane localization as the possible basis for these differences. In addition, the possibility that statin benefits on stroke reflect pleiotropic effects is reviewed. Finally, possible reasons for differences between the effects of atorvastatin metabolites and antioxidant vitamins are considered. PMID- 17126679 TI - Oxidative biomarkers in the diagnosis and prognosis of cardiovascular disease. AB - Oxidative damage to lipids and proteins is an important component of atherosclerotic cardiovascular disease (CVD). Studies of oxidation-related molecules are helping to define atherosclerotic mechanisms, and measurements of circulating levels of specific oxidant compounds may improve cardiovascular risk assessment. The present article reviews accumulating data of selected oxidative biomarkers that support their role in providing diagnostic and/or prognostic information. For example, plasma levels of the enzyme myeloperoxidase, which generates the strong oxidizing agent hypochlorous acid, have been found to be correlated with risk for myocardial infarction and endothelial dysfunction. Elevated levels of lipoprotein-associated phospholipase A(2) are associated with coronary artery disease (CAD) and stroke. Oxidized phospholipids play an important role in atherosclerosis. Recent studies measuring circulating levels of oxidized phospholipids have suggested a strong association with CAD, plaque disruption, and response to 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor ("statin") therapy. Isoprostanes correlate strongly with cardiovascular risk factors, but their role in risk prediction is less well defined. Future studies are expected to clarify the role of oxidative biomarkers in the diagnosis and prognosis of CVD and to determine their value in specific clinical populations. PMID- 17126681 TI - Patient safety: a view from the top. AB - The view of patient safety from the top encompasses vision, a long-range horizon, and fluency in the current state of system performance. Leadership in patient safety is an active process that demonstrates actions derived from safety as a value. It requires leaders who are accountable, responsible, knowledgeable, vigilant, and directly engaged. Patient safety is not a portfolio of work for leaders to preside over; rather, it is a core strategy of the organization and the health care industry that demands involvement. Patient safety is the work of leaders in this generation of health care: our watch, our time, and our work in achieving the delivery of harm-free care to children. PMID- 17126682 TI - The incidence of adverse events and medical error in pediatrics. AB - In its 2000 report, To Err is Human, the Institute of Medicine concluded that between 44,000 and 98,000 deaths per year occur in United States hospitals as a result of error. These data have resulted in calls for further research, regulatory interventions, third-party payer involvement, and health care organization initiatives to improve this situation. Studies of pediatric inpatients suggest that medication-related harm occurs at a rate as high as 11.1 per 100 admissions, and hospital-related harm occurs in high risk neonatal ICUs at a rate of 74 per 100 admissions. This article discusses differences between error and harm, methods used to measure harm, and available evidence that identifies the incidence of adverse events in pediatric inpatients and outpatients. PMID- 17126683 TI - Medical errors: understanding the parent's perspective. AB - Health care practitioners who advocate for full and open disclosure of medical errors often are met with opposition from legal advisors, insurance providers, hospital leadership, and colleagues. Although some progress has been made, a culture of fear around blame and retribution persists and continues to stymie the progression toward open discussion and disclosure of adverse events. The following case discussion addresses some common obstacles to disclosure of medical errors and reversals the potential for positive outcomes for patients and their families, hospital staff, and the health care system when those challenges are overcome. PMID- 17126684 TI - Disclosure of medical errors. AB - The 1999 release of the Institute of Medicine's document To Err is Human was akin to removing the lid of Pandora's box. Not only were the magnitude and impact of medical errors now apparent to those working in the health care industry, but consumers or health care were alerted to the occurrence of medical events causing harm. One specific solution advocated was the disclosure to patients and their families of adverse events resulting from medical error. Knowledge of the historical perspective, ethical underpinnings, and medico-legal implications gives us a better appreciation of current recommendations for disclosing adverse events resulting from medical error to those affected. PMID- 17126685 TI - Human factors engineering and patient safety. AB - The pediatric population has features different from those of adults and that are dynamic during the pediatric age range. Pediatric-specific issues result in potential risks for harm during medical care. Basic and applied human factors research has resulted in improvements in the performance of health adults and those adults who have functional limitations. Future work should focus on systematically understanding the human factors needs of children with the goal of redesigning systems of health care to optimize the safety of children and the performance of their care providers. PMID- 17126686 TI - Reliability science and patient safety. AB - Reliability is failure-free operation over time--the measurable capability of a process, procedure, or service to perform its intended function. Reliability science has the potential to help health care organizations reduce defects in care, increase the consistency with which care is delivered, and improve patient outcomes. Based on its principles, the Institute for Health care Improvement has developed a three-step model to prevent failures, mitigate the failures that occur, and redesign systems to reduce failures. Lessons may also be learned from complex organizations that have already adopted the principles of reliability science and operate with high rates of reliability. They share a preoccupation with failure, reluctance to simplify interpretations, sensitivity to operations, commitment to resilience, and underspecification of structures. PMID- 17126687 TI - The impact of fatigue on patient safety. AB - Modern medicine is founded on a culture of diligent, fatigued physicians. Fatigue is not desirable; however, the task of managing fatigue in health care professionals is complex and an ideal solution has not been described. Solutions need to integrate the immediate need for continued high-quality patient care, education of trainees, and the limited supply of health and human resources. Exploration of alternate scheduling models, broadened scope of practice, and new models of care delivery in demonstration projects or formal studies should be performed before widespread implementation. Appropriate evaluations are essential if well-meaning but larger scale errors in the name of patient safety are to be avoided. PMID- 17126688 TI - Medication errors in children. AB - Medication error is a major source of iatrogenic injuries in children. Dosing errors are the most common type of medication errors in pediatrics. Sicker patients in intensive care units and emergency departments are more often harmed by such errors. Strategies that have been found to be effective in reducing medication errors include the use of computerized physician order entry systems, preprinted order forms, and color-coded systems. Adopting the "systems approach" to medication errors is crucial to every health system where practitioners seek to enhance patient safety. PMID- 17126689 TI - Computerized provider order entry and patient safety. AB - Computerized provider order entry (CPOE) and clinical decision support (CDS) are advocated health care information technologies for improving patient safety through reduction and prevention of medication errors. CPOE and DCS target specific errors in medication processes, particularly in prescribing and ordering. These are of particular importance in pediatrics, because children may be more vulnerable to prescribing errors than adults. Studies of CPOE/CDS performed at academic medical centers have demonstrated their effectiveness in reducing medication process errors in pediatrics, but scant data so far show effects on health outcomes. CPOE/CDS adoption requires significant expertise in health care processes, information technology, and change management. Adoption is a high-cost, high-risk venture with political implications. PMID- 17126690 TI - Handoffs in care--can we make them safer? AB - In today's complex and rapidly changing health care environments, patient harm may result if important patient information is not communicated from one health care provider to another during handoffs in care. Issues involving communication, continuity of care, and care planning are cited as a root cause in more than 80% of reported sentinel events. In light of the inherent risks associated with handoffs in care, the use of strategies that reduce the impact of human factors on effective communication and standardize the communication process is essential to ensure appropriate communication patient information and that a plan of care is continued through the process. PMID- 17126691 TI - Racial/ethnic disparities and patient safety. AB - The number of racial/ethnic minority children will exceed the number of non Latino white children in the United States by 2030, but little is known about racial/ethnic disparities in patient safety, particularly when it comes to children. A literature review revealed several racial/ethnic disparities in pediatric patient safety. A new conceptual model is proposed for understanding racial/ethnic disparities in patient safety. Evidence suggests that improving patient safety for minority children could be achieved by routinely collecting and monitoring parental self-reported data on race/ethnicity, language, and English proficiency; enhancing the cultural competency of health care providers and staff; and providing adequate language services for all limited English proficient patients and their families. PMID- 17126692 TI - Patient safety: a view from Down Under. AB - The patient safety program at the Royal Children's Hospital, Melbourne has been running for more than 10 years. The hospital is in the State of Victoria, a state that has a long history of investigating child death, enacting legislation to reduce childhood injury, and providing a centralized tertiary care service. The close interaction between the hospital and the community provides opportunities for shared learning in pediatric patient safety to continually improve childhood outcomes. PMID- 17126693 TI - Tailoring the Institute for Health Care Improvement 100,000 Lives Campaign to pediatric settings: the example of ventilator-associated pneumonia. AB - In December 2004, the Institute for Healthcare Improvement launched a campaign to save 100,000 lives by implementing evidence-based interventions in six areas, five of which are relevant to children. Working collaboratively, the Child Health Corporation of America, National Associate of Children's Hospitals and Related Institutions, and National Initiative for Children's Health Care Quality provided a series of Web-enabled seminars on how the campaign initiatives might be adapted for pediatric settings. Ventilator-associated pneumonia (VAP) is an example of how interventions based on evidence in adult settings may need to be tailored in pediatric settings. The authors describe how assessing and implementing parts of the VAP bundle led to reduction in VAP in two children's hospitals. PMID- 17126694 TI - Blueprint for patient safety. AB - This article outlines a plan called the Blueprint for Patient Safety, which one organization has used to implement a comprehensive patient safety program. The blueprint starts with a graphic that depicts the basic objectives of patient safety and clarifies the sometimes misunderstood relationship between patient safety and more traditional quality-improvement activities. A second schematic outlines the key elements of the plan, including the concepts of continuous learning and the encompassing role of leadership. Finally, the 10 components of the plan and specific elements that could fall under each component are described. PMID- 17126695 TI - Addressing Africa. PMID- 17126696 TI - The ethics of premature delivery. PMID- 17126697 TI - Free the Benghazi six. PMID- 17126698 TI - Snake venom, vampire bat saliva, or rt-PA: time matters. PMID- 17126699 TI - England's national Teenage Pregnancy Strategy. PMID- 17126700 TI - Hospital admission for complications of unsafe abortion. PMID- 17126701 TI - The joint learning initiative on children and HIV/AIDS. PMID- 17126702 TI - From minuscule biomedical models to sexuality's depths. PMID- 17126703 TI - Alignment, harmonisation, and accountability in HIV/AIDS. PMID- 17126704 TI - What is publication? PMID- 17126705 TI - Research misconduct: learning the lessons. PMID- 17126706 TI - Julio Montaner: king of HAART. PMID- 17126707 TI - Neglected tropical diseases and HIV/AIDS. PMID- 17126708 TI - Neglected tropical diseases and HIV/AIDS. PMID- 17126709 TI - Global prevalence of meticillin-resistant Staphylococcus aureus. PMID- 17126711 TI - Altered fractionated radiotherapy in head and neck cancer. PMID- 17126712 TI - Altered fractionated radiotherapy in head and neck cancer. PMID- 17126713 TI - Parenteral nutrition in adults with a functional gastrointestinal tract. PMID- 17126715 TI - Will the new WHO growth references do more harm than good? PMID- 17126717 TI - Dapoxetine treatment of premature ejaculation. PMID- 17126718 TI - Of prazosin and purses. PMID- 17126719 TI - Intravenous ancrod for acute ischaemic stroke in the European Stroke Treatment with Ancrod Trial: a randomised controlled trial. AB - BACKGROUND: Intravenous tissue plasminogen activator is the only approved specific treatment for acute ischaemic stroke. Ancrod, a natural defibrinogenating agent from snake venom, has proved to have a favourable effect when given within 3 h after an acute ischaemic stroke. The European Stroke Treatment with Ancrod Trial was undertaken to assess the effects of ancrod when given within 6 h. METHODS: 1222 patients with an acute ischaemic stroke were included in this randomised double-blind placebo-controlled trial. Brain CT scans were done to exclude intracranial haemorrhages and large evolving ischaemic infarctions. Patients were randomly assigned ancrod (n=604) or placebo (n=618). The primary outcome was functional success at 3 months (survival, Barthel Index of 95 or 100, or return to prestroke level). The analysis was by intention-to treat. This trial is registered with ClinicalTrials.gov, trial number NCT00343174. FINDINGS: Functional success at 3 months did not differ between patients given ancrod (42%) and those given placebo (42%) (p=0.94, OR=0.99, 95% CI, 0.76-1.29). INTERPRETATION: On the basis of our findings, ancrod should not be recommended for use in acute ischaemic stroke beyond 3 h. PMID- 17126720 TI - Teenage conceptions, abortions, and births in England, 1994-2003, and the national teenage pregnancy strategy. AB - BACKGROUND: The aim of this study was to quantify the change in the number of conceptions and abortions among women younger than 18 years in England in relation to the government's national teenage pregnancy strategy. METHODS: We undertook geographic analysis of data for 148 top-tier local authority areas. The main outcomes were changes in under-18 conceptions, abortions, and births between the 5-year period before implementation of the strategy (1994-98) and the period immediately after implementation (1999-2003). FINDINGS: The number of teenage conceptions peaked in 1998, then declined after the implementation in 1999 of the teenage pregnancy strategy. Under-18 conception rates fell by an average of 2.0% (95% CI 1.8 to 2.2) per year between 1998 and 2003, below the rate needed to achieve the target of 50% reduction by 2010. The net change between 1994-98 and 1999-2003 was a fall in conceptions of 3.2% (2.6 to 3.9) or 1.4 per 1000 women aged 15-17 years, a rise in abortions of 7.5% (6.5 to 8.6) or 1.4 per 1000, and a fall in births of 10.6% (9.9 to 11.3) or 2.8 per 1000. The change in the number of conceptions was greater in deprived and more rural areas, and in those with lower educational attainment. The change was greater in areas where services and access to them were poorer, but greater where more strategy-related resources had been targeted. INTERPRETATION: The decline in under-18 conception and birth rates since 1998 and evidence that the declines have been greatest in areas receiving higher amounts of strategy-related funding provides limited evidence of the effect of England's national teenage pregnancy strategy. The full effect of local prevention will be clear only with longer observation, and substantial further progress is needed to remedy England's historically poor international position in teenage conceptions. PMID- 17126721 TI - Hospital admissions resulting from unsafe abortion: estimates from 13 developing countries. AB - BACKGROUND: Complications from unsafe abortion are believed to account for the largest proportion of hospital admissions for gynaecological services in developing countries. The WHO estimates that one in eight pregnancy-related deaths result from unsafe abortions. The social stigma and legal restrictions associated with abortion in many countries means that data on the magnitude of this problem are scarce; this article estimates the rate and numbers of hospital admissions resulting from unsafe abortions in developing countries to help quantify the problem. METHODS: National estimates of abortion-related hospital admissions in women aged 15-44 years were compiled for 13 developing countries: Africa (Egypt, Nigeria, and Uganda), Asia (Bangladesh, Pakistan, and the Philippines), and Latin America and the Caribbean (Brazil, Chile, Colombia, Dominican Republic, Guatemala, Mexico, and Peru). These data were combined with supplementary data from five countries in sub-Saharan Africa (Burkina Faso, Ghana, Kenya, Nigeria, and South Africa) to give estimates for the three world regions. FINDINGS: The annual hospitalisation rate varies from a low of about 3 per 1000 women in Bangladesh to a high of about 15 per 1000 in Egypt and Uganda. Nigeria, Pakistan, and the Philippines have rates of 4-7 per 1000, and two countries in Latin America with recent data have rates of almost 9 per 1000. In the developing world as a whole, an estimated five million women are admitted to hospital for treatment of complications from induced abortions each year. This equates to an average rate of 5.7 per 1000 women per year in all developing regions, excluding China. By comparison, in developed countries complications from abortion procedures or hospitalisation are rare. INTERPRETATION: These results help quantify the magnitude of the adverse health effects of unsafe abortion in developing countries and highlight the need for improved access to post-abortion care. The provision of abortion services is changing to include the drug misoprostol and this could reduce the severity of abortion complications and the number of women who are hospitalised. Researchers will need to monitor these changes to provide countries with up-to-date information on illness and death from unsafe abortion. Improved contraceptive services are necessary to prevent unintended pregnancy. However, increasing access to safe abortion services is the most effective way of preventing the burden of unsafe abortion, and remains a high priority for developing countries. PMID- 17126722 TI - Rectum compression by a gluteal lipoma. PMID- 17126723 TI - Acute myeloid leukaemia. AB - Acute myeloid leukaemia (AML) is a heterogeneous clonal disorder of haemopoietic progenitor cells and the most common malignant myeloid disorder in adults. The median age at presentation for patients with AML is 70 years. In the past few years, research in molecular biology has been instrumental in deciphering the pathogenesis of the disease. Genetic defects are thought to be the most important factors in determining the response to chemotherapy and outcome. Whereas significant progress has been made in the treatment of younger adults, the prospects for elderly patients have remained dismal, with median survival times of only a few months. This difference is related to comorbidities associated with ageing and to disease biology. Current efforts in clinical research focus on the assessment of targeted therapies. Such new approaches will probably lead to an increase in the cure rate. PMID- 17126724 TI - Unsafe abortion: the preventable pandemic. AB - Ending the silent pandemic of unsafe abortion is an urgent public-health and human-rights imperative. As with other more visible global-health issues, this scourge threatens women throughout the developing world. Every year, about 19-20 million abortions are done by individuals without the requisite skills, or in environments below minimum medical standards, or both. Nearly all unsafe abortions (97%) are in developing countries. An estimated 68 000 women die as a result, and millions more have complications, many permanent. Important causes of death include haemorrhage, infection, and poisoning. Legalisation of abortion on request is a necessary but insufficient step toward improving women's health; in some countries, such as India, where abortion has been legal for decades, access to competent care remains restricted because of other barriers. Access to safe abortion improves women's health, and vice versa, as documented in Romania during the regime of President Nicolae Ceausescu. The availability of modern contraception can reduce but never eliminate the need for abortion. Direct costs of treating abortion complications burden impoverished health care systems, and indirect costs also drain struggling economies. The development of manual vacuum aspiration to empty the uterus, and the use of misoprostol, an oxytocic agent, have improved the care of women. Access to safe, legal abortion is a fundamental right of women, irrespective of where they live. The underlying causes of morbidity and mortality from unsafe abortion today are not blood loss and infection but, rather, apathy and disdain toward women. PMID- 17126725 TI - Assessing the effect of the 2001-06 Mexican health reform: an interim report card. AB - Since 2001, Mexico has been designing, legislating, and implementing a major health-system reform. A key component was the creation of Seguro Popular, which is intended to expand insurance coverage over 7 years to uninsured people, nearly half the total population at the start of 2001. The reform included five actions: legislation of entitlement per family affiliated which, with full implementation, will increase public spending on health by 0.8-1.0% of gross domestic product; creation of explicit benefits packages; allocation of monies to decentralised state ministries of health in proportion to number of families affiliated; division of federal resources flowing to states into separate funds for personal and non-personal health services; and creation of a fund to protect families against catastrophic health expenditures. Using the WHO health-systems framework, we used a wide range of datasets to assess the effect of this reform on different dimensions of the health system. Key findings include: affiliation is preferentially reaching the poor and the marginalised communities; federal non social security expenditure in real per-head terms increased by 38% from 2000 to 2005; equity of public-health expenditure across states improved; Seguro Popular affiliates used more inpatient and outpatient services than uninsured people; effective coverage of 11 interventions has improved between 2000 and 2005-06; inequalities in effective coverage across states and wealth deciles has decreased over this period; catastrophic expenditures for Seguro Popular affiliates are lower than for uninsured people even though use of services has increased. We present some lessons for Mexico based on this interim evaluation and explore implications for other countries considering health reforms. PMID- 17126726 TI - Unexplained choroidal embolisation: remember the antiphospholipid syndrome. PMID- 17126727 TI - Proceedings of the 9th European symposium on calcium-binding proteins in normal and transformed cells. Strasbourg, France, 19-22 July 2006. PMID- 17126728 TI - NaCl stress effects on enzymes involved in nitrogen assimilation pathway in tomato "Lycopersicon esculentum" seedlings. AB - Tomato plants (Lycopersicon esculentum Mill, cv. Chibli F1) grown for 10 days on control medium were exposed to differing concentrations of NaCl (0, 25, 50, and 100mM). Increasing salinity led to a decrease of dry weight (DW) production and protein contents in the leaves and roots. Conversely, the root to shoot (R/S) DW ratio was increased by salinity. Na(+) and Cl(-) accumulation were correlated with a decline of K(+) and NO(3)(-) in the leaves and roots. Under salinity, the activities of nitrate reductase (NR, EC 1.6.6.1) and glutamine synthetase (GS, EC 6.3.1.2) were repressed in the leaves, while they were enhanced in the roots. Nitrite reductase (NiR, EC 1.7.7.1) activity was decreased in both the leaves and roots. Deaminating activity of glutamate dehydrogenase (GDH, EC 1.4.1.2) was inhibited, whereas the aminating function was significantly stimulated by salinity in the leaves and roots. At a high salt concentration, the nicotinamide adenine dinucleotide reduced (NADH)-GDH activity was stimulated concomitantly with the increasing NH(4)(+) contents and proteolysis activity in the leaves and roots. With respect to salt stress, the distinct sensitivity of the enzymes involved in nitrogen assimilation is discussed. PMID- 17126729 TI - Antioxidative response of ascorbate-glutathione pathway enzymes and metabolites to desiccation of recalcitrant Acer saccharinum seeds. AB - Ascorbate-glutathione systems were studied during desiccation of recalcitrant seeds of the silver maple (Acer saccharinum L.). The desiccated seeds gradually lost their germination capacity and this was strongly correlated with an increase in electrolyte leakage from seeds. Simultaneously the increase of reactive oxygen species (ROS) (superoxide radical - O(2)(-*) and hydrogen peroxide - H(2)O(2)) production was observed. The results indicate that remarkable changes in the concentrations and redox status of ascorbate and glutathione occur in embryo axes and cotyledons. After shedding, concentrations of ascorbic acid (ASA) and the reduced form of glutathione (GSH) are higher in embryo axes than in cotyledons and their redox status is high in both embryo parts. Cotyledons in freshly shed seeds are devoid of GSH. At the first stages of desiccation, up to a level of 43% of moisture content, ASA content in embryo axes and GSH content in cotyledons increased. Below this level of moisture content, the antioxidant contents as well as their redox status rapidly decreased. The enzymes of the ascorbate-glutathione pathway: ascorbate peroxidase (APX) (EC 1.11.1.11), monodehydroascorbate reductase (MR) (EC 1.6.5.4), dehydroascorbate reductase (DHAR) (EC 1.8.5.1) and glutathione reductase (GR) (EC 1.6.4.2) increased their activity during desiccation, but mainly in embryonic axes. The changes are probably required for counteracting the production of ROS during desiccation. The relationship between ascorbate and glutathione metabolism and their relevance during desiccation of recalcitrant Acer saccharinum seeds is discussed. PMID- 17126730 TI - A major factor in gravitropism in radish hypocotyls is the suppression of growth on the upper side of hypocotyls. AB - The changes in length on the two opposite sides of etiolated radish (Raphanus sativus) hypocotyls prior to, and following gravitropic stimulation, were measured using an infrared-imaging system. It was observed that the growth suppression on the upper side began first at least 10 min after the onset of gravitropic stimulation, and after 30 min the acceleration in growth on the lower side started. The gravitropic curvature was steadily induced from 10 min. When radish hypocotyls were switched from a vertical to horizontal position for different durations and then replaced to the vertical position, the growth suppression on the gravity-stimulated (upper) side was observed in all cases, but the acceleration in growth on the opposite (lower) side appeared only in continuously gravity-stimulated seedlings, although it occurred later than the growth suppression on the upper side. These results suggest that the suppression in growth on the upper side of the hypocotyls is a direct effect of gravitropic stimulation, but not the acceleration on the lower side. When 4-methylthio-3 butenyl isothiocyanate (4-MTBI), which has an inhibitory activity against radish hypocotyl growth, was applied on the one side of radish hypocotyls and then the 4 MTBI-applied side or opposite side was placed in a horizontal position, the former showed greater bending than the control, suggesting that the growth suppression on the upper side is enhanced and maintained with MTBI application there. In the latter case, the seedlings showed less bending than the control, suggesting a decrease in growth on the lower side with MTBI application. All the results suggest that gravitropism of radish hypocotyls may be caused by an increase in growth-inhibiting substance(s) induced with gravitropic stimulation in the upper side, inducing growth inhibition there. PMID- 17126731 TI - Distribution of UV-shielding of the epidermis of sun and shade leaves of the beech (Fagus sylvatica L.) as monitored by multi-colour fluorescence imaging. AB - Plants can protect against damaging ultraviolet (UV) radiation by accumulating UV absorbing substances in the epidermis of the leaves. Sun and shade leaves of a free standing beech tree (Fagus sylvatica L.) were studied for the differences in UV-shielding of the epidermis by means of multi-colour fluorescence images taken with UV and blue excitation. The distribution of the fluorescence intensity was detected over intact leaves in the emission maxima in the blue at 440 nm (F440), in the green at 520 nm (F520), in the red at 690 nm (F690) and in the far red at 740 nm (F740). Images of the logarithmic ratio between F690 excited in the blue and the UV (log ((B)F690/(UV)F690)) were calculated representing the relative absorption of UV in the epidermis and thus the degree of UV-shielding. It was found that UV-shielding is stronger for sun leaves than for shade leaves and better for the upper (adaxial) leaf side than for the lower (abaxial) leaf side of both leaf types. Within one leaf the highest value for the ratio log ((B)F690/(UV)F690) and thus the highest UV-shielding was found at the leaf rim which in broad leaves contains young tissue. PMID- 17126732 TI - Influence of NaCl and mannitol on peroxidase activity and lipid peroxidation in Centaurea ragusina L. roots and shoots. AB - Centaurea ragusina L. is a Croatian endemic plant species growing on cliffs above the Adriatic Sea, but there is no information about its physiological behavior or stress tolerance. To investigate the response of C. ragusina plants to salinity and drought, we have analysed soluble peroxidase (POD; EC 1.11.1.7) activity, anionic isoperoxidase pattern, levels of malondialdehyde (MDA) and hydrogen peroxide in C. ragusina plants exposed to these stresses. Rooted plantlets grown on MS 1/2 nutrient medium supplemented with mannitol (300 mM) and different concentrations of NaCl (150, 300, 450 or 600 mM) were harvested after 5, 10 and 15 days. Both osmotic treatments significantly increased MDA and hydrogen peroxide contents in C. ragusina shoots after 10 days of stress, while in roots these parameters showed no significant difference compared to control in overall. POD activity of salt-stressed plants changed with respect to different saline treatments and plant organs - in shoots enzymatic activity markedly increased in response to lower saline treatments, especially 300 mM NaCl; otherwise it was similar as in control plants while in roots of plants grown under 450 and 600 mM NaCl it significantly decreased. Drought increased POD activity of both shoots and roots especially after 10 days of experiment. Generally, change in the POD isoenzyme pattern of treated plants was in accordance with the activity change in time. Several POD isoforms (P3, P4 and P9) were specifically induced by salinity and drought. PMID- 17126733 TI - Local NO3- or NH4+ supply modifies the root system architecture of Cedrus atlantica seedlings grown in a split-root device. AB - To study the effects of local nitrate or ammonium supply on the architecture of the Cedrus atlantica root system, cedar seedlings were grown in split-root boxes in a growth chamber. In each box-compartment, roots were fertilized with a solution containing nitrogen, either as nitrate [Ca(NO(3))(2)] or ammonium (NH(4)Cl), supplied at 0.1 or 5.0mM. For each seedling, the shoot growth was measured twice a week for 3 months. The root system architecture was also recorded twice a week by tracing the root elongation through the transparent face of the root observation boxes. The apical diameter of the tap-root relay and that of a representative sample of lateral roots were recorded once a month using a monocular magnifier. The increase of ammonium or nitrate concentration in the nutrient solution has significantly enhanced the production of lateral roots on the tap-root relay. After 90 days of culture, percentages of short lateral roots obtained with nitrate were higher than those obtained using ammonium. A preferential carbon allocation to the shoots was also obtained with an increasing nitrogen supply. Until the 40th day of culture, the elongation of lateral roots was similar for all treatments and ranged from 0.25 to 0.5 cm day(-1). From the 40th day to the 95th day, significant differences were observed between the compared modes and maximum elongation rates were obtained with 5mM NH(4)(+) (2.18 cm day(-1)) and 5mM NO(3)(-) (1.18 cm day(-1)). Local applications of nitrate and ammonium at a low or a high concentration had local effects on elongation and branching of the root system in the fertilized compartment. Contrasting effects of ammonium and nitrate were observed on the apical diameter of tap-roots and lateral roots. The root-split culture device confirmed that nitrate had local effects on the architecture of the C. atlantica root system. PMID- 17126734 TI - Quantitative determination of callose in tree roots. AB - The formation of callose in tree roots has been suggested as a physiological indicator of aluminum (Al) toxicity. Quantifying callose in the roots in forest soils, however, is hampered by the presence of autofluorescent materials in the roots that disturb the measurement of callose by fluorescence spectrophotometry. Tannins in the roots cause these measurement problems. Here we report on the measurement of callose in the root apices of European chestnut (Castanea sativa) seedlings collected in an acidified forest soil. The callose was quantified with a modified protocol which included three washing steps with polyvinylpolypyrrolidone (PVPP) before the callose was extracted. This procedure reduced the autofluorescence by about 50%. With the use of water or ethanol alone, callose could be measured in only about 15% of the root samples, whereas with the use of PVPP callose could be determined in 95% of the samples. This improved method could help to evaluate the effects of Al toxicity on tree roots grown in forest soils, where callose is detected as a physiological indicator. PMID- 17126736 TI - Introduction to photodynamic therapy: early experience. AB - Photodynamic therapy (PDT) has become one of the newest and most exciting therapies being used by dermatologic surgeons from all around the world. The earliest uses of PDT can be traced to the early 1900s. This article traces the history of PDT through the discovery of the first topical photosensitizer by Kennedy and his colleagues in 1990 to its uses today in everyday dermatology practices. The two commercially available photosensitizers are reviewed and their primary uses are summarized. PDT is a useful modality that is finding new and exciting niches in today's dermatologic world. PMID- 17126737 TI - Aminolevulinic acid photodynamic therapy for skin cancers. AB - Aminolevulinic acid photodynamic therapy (ALA-PDT) is an effective and noninvasive therapy for superficial basal cell carcinoma (BCC) and Bowen's disease. It also may have a role in the treatment of nodular BCC and other cutaneous malignancies, including localized cutaneous lymphomas. ALA-PDT offers multiple advantages over traditional treatments, including little to no scarring, excellent cosmetic results, and the ability to treat multiple lesions simultaneously. It is not an effective therapy for aggressive subtypes of BCC or for invasive squamous cell carcinoma. Finally, ALA-PDT may be a useful way to prevent new skin cancers in certain high-risk patients. PMID- 17126738 TI - Photodynamic therapy for actinic keratoses. AB - Actinic keratoses (AKs) are one of the most common conditions that are treated by dermatologists and they have the potential to progress to squamous cell carcinoma if left untreated. Photodynamic therapy (PDT) has emerged as a novel and versatile method of treating those lesions. Topical preparations of aminolevulinic acid and methyl aminolevulinate are commercially available photosensitizers, and numerous light sources may be used for photoactivation. This article focuses on practical aspects of PDT in the treatment of AKs, outcomes of relevant clinical trials, and special applications of PDT in transplant recipients and other who are predisposed to AK formation. Step-by-step descriptions of PDT sessions are presented. PMID- 17126739 TI - Aminolevulinic acid photodynamic therapy for actinic keratoses/actinic cheilitis/acne: vascular lasers. AB - The combination of newer laser and light sources, the long-pulsed pulsed dye laser (LP PDL) and intense pulsed light, with topical aminolevulinic acid photodynamic therapy (PDT) has achieved enhanced efficacy and rapid treatment and recovery, while diminishing unwanted side effects. In particular, LP PDL PDT has been shown to be safe and effective in the treatment of actinic keratoses, actinic cheilitis, photodamage, and acne vulgaris with minimal discomfort, rapid treatment and recovery, and excellent posttreatment cosmesis. PMID- 17126740 TI - Aminolevulinic acid-photodynamic therapy for photorejuvenation. AB - Light therapy is widely used in dermatology. An enhancement to his therapy is the use of a photosensitizing medication along with light therapy, known as photodynamic therapy (PDT). PDT is primarily used for the treatment of precancerous lesions, acne vulgaris, and nonmelonoma skin cancer, and has also been shown to improve the appearance of photodamaged skin. This article reviews the available literature for the treatment of photodamaged skin using PDT along with recent advancements in PDT for various dermatologic indications. PMID- 17126741 TI - The use of photodynamic therapy for treatment of acne vulgaris. AB - Current topical and most oral therapies for acne vulgaris have limited efficacy, especially in moderate to severe cases. Photodynamic therapy with 5 aminolevulinic acid and recently methyl aminolevulinate has been shown to be a safe and effective modality for the treatment of acne vulgaris. Consensus guidelines suggest that 30 to 60 minutes is sufficient 5-aminolevulinic acid contact time before photoactivation with blue light, red light, yellow light, broadband light, halogen, or pulsed dye laser devices. An average of three treatment can yield significant long-term improvement. PMID- 17126742 TI - Aminolevulinic acid photodynamic therapy for sebaceous gland hyperplasia. AB - Current therapies for sebaceous hyperplasia (SH) have a high risk for adverse effects and recurrence of treated lesions. The theoretic basis for the treatment of SH by photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA) has been established. Studies show that 1 hour is sufficient ALA incubation time to achieve clearance, and ALA-induced protoporphyrin IX may be activated with a 585 nm pulsed dye laser device, blue light source, or an intense pulsed light device. Complete clearance may be achieved with one to six treatments; however, long-term recurrence rates are not established. ALA PDT is a safe and effective modality for the treatment of SH lesions of all sizes. PMID- 17126743 TI - Aminolevulinic acid photodynamic therapy for hidradenitis suppurativa. AB - Hidradenitis suppurative (HS) is a chronic, often suppurative chronic dermatologic disorder that is often misdiagnosed. It has presented therapeutic challenges for decades since its first descriptions in the medical literature in 1854. HS is an apocrine disorder with high prevalence, which has a genetic predisposition. Numerous treatment options have been described, some to control the acute disease process and others to control its long-term consequences. The use of photodynamic therapy (PDT) recently has been described as a treatment option for intractable HS. The clinical trials that are associated with PDT and HS are reviewed, and thoughts about the probably mechanisms of actions of this therapy are given. PDT should be considered in patients who have severe HS. PMID- 17126744 TI - Treatment of verrucae vulgaris and molluscum contagiosum with photodynamic therapy. AB - The use of 5-aminolevulinc acid photodynamic therapy in the treatment of recalcitrant verrucae vulgaris and in the treatment of recalcitrant molluscum contagiosum has been shown to substantially reduce the lesion count and severity. Clinical research published in the medical literature and personal experience of both of the authors support the use of 5-aminolevulinc acid photodynamic therapy in appropriate individuals with recalcitrant verrucae vulgaris and molluscum contagiosum lesions. This article reviews the medical literature and identifies the various lasers and lights sources used to treat these conditions. PMID- 17126745 TI - Methyl aminolevulinate: actinic keratoses and Bowen's disease. AB - Topical photodynamic therapy (PDT) using the methyl ester of 5-aminolaevulinic acid (MAL) is an effective therapy for actinic keratoses and Bowen's disease. Thin and moderate thickness facial actinic keratoses respond best, with clearance rates equivalent or superior (depending on protocol) to current therapy, and with notably superior cosmetic outcome. Patients with areas of field cancerization and organ transplant recipients may particularly benefit from topical MA-PDT. The response rate of Bowen's disease to MAL-PDT is also at least equivalent to cryotherapy and 5-fluorouracil, again with superior cosmesis. Patients with large or multiple lesions of Bowen's disease or those in whom standard therapy, including surgery, is relatively contraindicated may particularly benefit from PDT. PMID- 17126746 TI - Methyl aminolevulinate-photodynamic therapy for basal cell carcinoma. AB - Basal cell carcinomas (BCCs) are the most common malignant tumors of the skin. Treatment of BCCs should be chosen according to clinical type, tumor size, and location. Methyl aminolevulinate (MAL) photodynamic therapy (PDT) has the potential to become a therapy with equal effectiveness to classical therapeutic modalities with an excellent cosmesis, but without complications like scar formation, requirement for grafts, need of repetitive treatments over longer time periods, or pigmentary changes. MAL is licensed in Europe, Australia, New Zealand, and Brazil for the treatment of actinic keratoses, Bowen's disease, and nodular and superficial BCC. Conclusions are drawn from extensive studies in past years using MAL-PDT for both nodular and superficial BCCs. PMID- 17126747 TI - Chemopreventative thoughts for photodynamic therapy. AB - The concept of skin cancer prevention with photodynamic therapy has evolved over the past few years to include large surface application of aminolevulinic acid or methyl aminolevulinate followed by light exposure to prevent the development of new lesions. Pre-clinical studies using various mouse models have shown that large surface photodynamic therapy can prevent the appearance of actinic keratoses, squamous cell carcinomas, and basal cell carcinomas. Recent clinical studies also suggest that large surface photodynamic therapy can prevent the development of actinic keratoses and possibly skin cancer. PMID- 17126748 TI - Photodynamic therapy: other uses. AB - Mainstream uses for photodynamic therapy (PDT) in dermatology include nonmelanoma skin cancer and its precursors, acne vulgaris, photorejuvenation, and hidradenitis suppurativa. Many other dermatologic entities have been treated with PDT, including psoriasis, lichen planus, lichen sclerosus, scleroderma, cutaneous T cell lymphoma, alopecia areata, verruca vulgaris, Darier's disease and tinea infections. Nondermatologic applications include anal and vulva carcinoma, palliation of metastatic breast cancer to skin, Barrett's esophagus, and macular degeneration of the retina. PDT also has found to be useful in immunologic and inflammatory disorders, neoplasias other than skin cancer, and infections. The ability of this treatment to hone in on dysplastic epithelial and endothelial cells while retaining viability of surrounding tissue is its key feature because this leads to specific tumor destruction with cosmesis and function of the target organ intact. PMID- 17126749 TI - Incorporating photodynamic therapy into a medical and cosmetic dermatology practice. AB - Effective in the treatment of a growing variety of skin conditions, photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA) is rising rapidly in popularity among both practitioners and patients, despite the fact that many applications are not yet cleared by the Food and Drug Administration. Because of its versatility, safety, efficacy, cosmetic benefits, and potential financial advantages, ALA-PDT may soon be a mainstay in many clinical settings. This article provides an overview of this easy-to-use treatment modality and a guide to implementing ALA-PDT into practice, including pretreatment and post-treatment protocols and guidelines for managing side effects. PMID- 17126750 TI - Photodynamic therapy in dermatology: the next five years. AB - Photodynamic therapy (PDT) has had a rapid insurgence into the dermatologic armamentarium of clinicians from all over the world. Its use in actinic keratoses and in non-melanoma skin cancers is well described. Additional uses, including its use in the treatment of photorejuvenation and acne vulgaris and related disorders, are gaining more widespread use. More recent descriptions, including its potential use as a chemoprevention agent, warrant further investigations and discussions. PDT is a global treatment modality and physicians and investigators must view the therapy as such, and use the best photosensitizer for the best indication, and continue to evaluate new approaches to maximize the use of PDT in dermatology. PMID- 17126752 TI - Epidemiology, pathophysiology, and prognosis of heart failure in the elderly. AB - Heart failure (HF) represents the quintessential disorder of cardio-vascular aging, reflecting the convergence of age-related changes in the cardiovascular system and other organ systems and the increasing prevalence of cardiovascular diseases at older age. The prevalence and incidence of HF increase progressively with advancing age, and HF imposes an enormous burden on society in mortality,morbidity, and associated health care costs. Despite major advances in treatment of HF over the last 25 years, the prognosis re-mains poor, with median survival rates of less than 5 years in older adults. As the population ages, it is anticipated that the number of older people with HF will increase dramatically over the next several decades. PMID- 17126753 TI - Clinical manifestations, diagnostic assessment, and etiology of heart failure in older adults. AB - Aging is characterized by heterogeneity, both in health and in dis-ease. Older adults who have heart failure (HF) often have a typical and delayed clinical manifestations and many have diastolic HF. The assessment and management of HF in older adults may be simplified by a 5-step process called DEFEAT HF: (1) Establish a clinical Diagnosis of HF; (2) Establish an Etiology for HF, preferably in collaboration with a cardiologist; (3) Determine Fluid status and achieve euvolemia; (4) Determine left ventricular Ejection frAction; and (5) Provide evidence-based Therapy. PMID- 17126754 TI - Role of echocardiography in the diagnostic assessment and etiology of heart failure in the elderly--opacify, quantify, and rectify. AB - Echocardiography offers comprehensive, noninvasive, and relatively inexpensive tools for diagnosing cardiac pathology in the elderly. With an organized approach using two-dimensional echocardiography and Doppler echocardiography, clinicians can determine the systolic and diastolic left ventricular performance;estimate the cardiac output, pulmonary artery, and ventricular fill-ing pressures; and identify surgically correctable valve disease.Meanwhile, real-time three dimensional echocardiography provides unprecedented volume data to quantify the left ventricular status. Tissue Doppler-derived myocardial velocity and strain imaging data provide extremely fine details about the regional variations in myocardial synchrony and predict responders to cardiac resynchronization therapy. Thus, echocardiographic tools provide the basis for determining when to attempt to rectify the left ventricular dysfunction with strategically placed, biventricular pace-maker leads. PMID- 17126755 TI - Treatment of heart failure with abnormal left ventricular systolic function in the elderly. AB - This article summarizes the four stages of heart failure (HF) as defined by the American College of Cardiology and the American Heart Association and discusses the treatments for elderly patients with HF and abnormal left ventricular systolic function. The article explains the important role of diuretics, the first-line drugs in the treatment of older patients with HF and volume overload. Other treatments described include angiotensin-converting enzyme inhibitors, angiotensin-receptor blockers, beta-blockers, aldosterone antagonists, isosorbide dinitrate plus hydralazine, digoxin, and calcium channel blockers. The article explains the role each of these plays and reports on studies that have examined and compared various treatments. PMID- 17126756 TI - Diastolic heart failure in the elderly. AB - Most elderly patients, particularly women, who have heart failure have a normal ejection fraction. Patients who have this syndrome have severe symptoms of exercise intolerance, frequent hospitalizations, and increased mortality. The pathophysiology and treatment are not well defined. Control of systemic hypertension may be a key to prevention and treatment. Several large trials of specific agents are currently underway. PMID- 17126757 TI - Use of diuretics in the treatment of heart failure in the elderly. AB - Diuretics are tools of considerable therapeutic importance. First, they effectively reduce blood pressure, while at the same time decreasing the morbidity and mortality associated with hypertension. Diuretics are currently recommended as first-line therapy for the treatment of hypertension. In addition, they remain an important component of heart failure therapy, in that they improve the symptoms of congestion, which typify the more advanced stages of heart failure. This article reviews the mode of action of the various diuretic classes and the physiologic adaptations that follow; sets up the basis for their use in the treatment of volume-retaining states, particularly as applies to the elderly; and reviews diuretic-related side effects that are normally encountered. PMID- 17126758 TI - Heart-failure-complicating acute myocardial infarction. AB - This article addresses issues related to acute myocardial infarction(MI) complicated by heart failure, particularly in elderly patients. Findings have shown that acute MI complicated by congestive heart failure (CHF) is associated with a high mortality, and that women with acute MI are more likely to be older and to develop CHF than men with acute MI. In general, management of CHF complicating acute MI is similar in older and younger patients. Actions discussed include hemodynamic monitoring; the administration of oxygen; and the use of morphine, diuretics, nitroglycerin,angiotensin-converting enzyme inhibitors, angiotensin II receptor blockers, spironolactone, beta-blockers, calcium channel blockers, magnesium, digoxin, and positive inotropic drugs. The article also discusses measures for treating arrhythmias and for diagnosing mechanical complications. PMID- 17126759 TI - Inotropic drugs and neurohormonal antagonists in the treatment of HF in the elderly. AB - HF (HF) is the most common reason for hospital admission among individuals over age 65 years and results in more than 1 million admissions each year. The overall annual death rate for HF is approximately 20%. HF results from decreased contractile function of the heart, and neurohormonal dysregulation plays a major part in the morbidity and mortality of the heart. The purpose of this article is to review recent studies on inotropic drugs and neurohormonal antagonists used in the treatment of patients who have HF, especially the elderly. PMID- 17126760 TI - Interventional therapies for heart failure in the elderly. AB - The aging of a population replete with risk factors for heart failure(HF) (coronary heart disease, diabetes, and hypertension) coupled with a declining age adjusted mortality rate for coronary artery and hypertensive heart diseases has created, and will continue to create, a literal explosion in the prevalence of HF in the United States. Despite advances in maximal medical therapy, however, most patients who have symptomatic HF can expect functional impairment, interludes of worsening symptomatology, and a shortened lifespan. This article updates the use of interventional therapies for the treatment of elderly patients who have HF caused by coronary artery disease, valvular heart disease, congenital heart disease, myocardial disease, and renal vascular disease. PMID- 17126761 TI - Surgical treatment of heart failure in the elderly. AB - Heart failure (HF) in elderly patients who may benefit from surgical therapy is usually secondary to ischemic or valvular heart disease. When referring such patients for surgery, life expectancy, along with the expectations of the patient and family with regard to the surgical treatment, must be considered. The goals of cardiac surgery in this patient population are to maintain or improve cardiac function, decrease HF episodes, reduce hospital admissions,and improve functional class. Safer surgical techniques developed during the last two decades have allowed high-risk patients well into their 80s to undergo complex cardiac operations with decreasing morbidity and mortality. Successful surgical intervention often leads to a more productive and independent life for elderly patients who have HF. PMID- 17126762 TI - Cardiac resynchronization therapy for treatment of heart failure in the elderly. AB - Congestive heart failure affects 5 million patients each year and is more common in elderly patients. Cardiac resynchronization therapy (CRT) or biventricular pacing is the simultaneous pacing of both ventricles, an approach that can improve symptoms and survival in some heart failure (HF) patients. Although no randomized controlled trial has evaluated CRT specifically in elderly patients,many of the patients included in the landmark CRT trials were aged 70 and older. Data suggest that CRT is a safe and efficacious treatment of HF in geriatric patients. PMID- 17126763 TI - Treatment of arrhythmias and use of implantable cardioverter-defibrillators to improve survival in elderly patients with cardiac disease. AB - Patients who have left ventricular dysfunction and heart failure(HF) are at high risk for ventricular tachyarrhythmias and sudden cardiac death. Randomized clinical trials have demonstrated that pharmacologic management with antiarrhythmic drugs has limited efficacy for the prevention of arrhythmic mortality in this high-risk population, whereas implantation of a defibrillator was shown to be associated with a significant survival benefit. Data on the efficacy of defibrillator therapy in elderly patients, in whom comorbidities are common, are limited. In this article we outline current information on therapeutic modalities for the prevention of arrhythmic mortality in elderly patients who have left ventricular dysfunction and HF, focusing mainly on data on the benefit of device therapy in the older age group. PMID- 17126764 TI - Exercise therapy for elderly heart failure patients. AB - Exercise intolerance is one of the hallmark symptoms of heart failure (HF). Exercise training in this condition was not explored as a therapeutic modality until the past two decades. In fact, bed rest was promoted as a treatment for certain forms of HF in the 1970s. Over the past two decades, however, data have shown convincingly that exercise training can be implemented safely in patients who have HF, with significant improvement in functional capacity, various other physiologic variables, and quality of life. This article reviews the accumulated literature in this area, with specific emphasis on the effects of exercise training in older patients who have HF, a subset that may have the most to gain from such training. PMID- 17126765 TI - End-of-life care in the treatment of heart failure in the elderly. AB - Much of the literature dedicated to the topic of medical care of dying patients has revolves around terminal care provided to patients who have neoplastic diagnoses. Heart failure (HF) presents its own unique challenges to the clinician. This article focuses on specific clinical recommendations and an analysis of some of the ethical issues involved in the provision of care to elderly patients in the terminal stages of HF. PMID- 17126766 TI - Normal weight obese (NWO) women: an evaluation of a candidate new syndrome. AB - BACKGROUND AND AIMS: Obesity, an independent risk factor for cardiovascular disease (CVD), has been associated with the early development of coronary atherosclerosis in adolescents and young men. A subset of metabolically obese but normal weight individuals was identified, with potentially increased risks for development of the metabolic syndrome despite their normal body mass index. We determined the relationship among body fat distribution and selected CVD risk factors to distinguish normal weight obese from controls with normal metabolic profiles. METHODS AND RESULTS: We analysed anthropometric variables, body composition by DXA, RMR by indirect calorimetry and bioumoral variables of 74 clinically healthy Caucasian Italian women. Significant differences were observed in the biochemical HDL-chol values between NWO and controls and pre-obese-obese. Significant correlations were found among cardiovascular risk indexes, LEAN of the right part of the trunk and TC/HDL (R=-0.69, p<0.001) and LDL/HDL (R=-0.72, p<0.001), and LEAN and RMR (R=0.44, p=0.022) of NWO women. CONCLUSIONS: In normal weight obese women the cardiovascular risk indexes are related to metabolic variables and to body fat mass distribution. NWO individuals showed a relationship between the decrease in LEAN of the left leg and an increase in CVD risk factors. We suggest that LEAN distribution seems to be a potential predictor of CVD. PMID- 17126767 TI - The Mediterranean and CHO diets decrease VCAM-1 and E-selectin expression induced by modified low-density lipoprotein in HUVECs. AB - BACKGROUND AND AIM: The oxidative modifications of low-density lipoprotein (LDL) are crucial for the atherosclerosis process. The aim of this study was to determine if the minimally modified LDL, obtained after the ingestion of three different diets, produce differential effects on the vascular cell adhesion molecule-1 (VCAM-1) and E-selectin expression in human umbilical endothelial cells (HUVECs). METHODS AND RESULTS: Twenty healthy young males were exposed to three dietary periods. Each period lasted four weeks. During the first period, all subjects consumed a saturated fat (SFA) enriched diet (38% fat, 20% SFA). The second and third dietary periods were administered following a randomized crossover design: a low fat high carbohydrates diet (CHO diet) and a Mediterranean diet. LDL particles, isolated during each dietary period, were oxidized by exposure to UV light and incubated for 48 h with HUVEC. Thereafter, 100 U/mL of TNF-alpha was added and incubation continued for 6 h. Cellular ELISA determined adhesion molecules expression. Lag time, propagation rate and total amounts of formed conjugated dienes were calculated in LDL incubated with 10mumol/L Cu(2+). When compared to the SFA diet, LDL isolated from the Mediterranean and CHO diets induced a lower expression of VCAM-1 and E-selectin in HUVECS (P<0.007). There were no differences between both lipid lowering diets. However, lag time of LDL from the Mediterranean diet was higher than with the CHO diet (P<0.042). This parameter was inversely correlated with E-selectin expression (r=-0.497; P<0.04). CONCLUSION: Our results suggest that both the Mediterranean and CHO diets may decrease the pro-inflammatory environment induced by modified LDL in endothelial cells. PMID- 17126768 TI - Effect of Lactobacillus fermentum on serum lipids in subjects with elevated serum cholesterol. AB - BACKGROUND AND AIMS: There is increasing interest in the use of natural therapies to reduce elevated low density lipoprotein (LDL) cholesterol. This study assessed the effects of PCC Lactobacillus fermentum on LDL cholesterol and other lipid fractions. METHODS AND RESULTS: This was a single centre, double blind, placebo controlled, parallel design trial in volunteers having total cholesterol > or =4 mmol/L. Subjects (n=46) were randomised to receive either Lactobacillus fermentum 2 capsules twice daily (each capsule containing 2 x 10(9) colony forming units) or matching placebo for a period of 10 weeks. Main outcome measures were percentage changes in LDL cholesterol and other lipids, changes in liver enzymes and other safety tests. Two subjects withdrew early in the study, 1 for personal reasons and 1 because of bowel discomfort. Three other subjects experienced some bowel discomfort but still completed the study. LDL cholesterol showed a modest downward trend on both Lactobacillus fermentum and placebo of 7.0% and 5.2% respectively. This trend did not reach statistical significance over time, nor was there a significant difference between the treatment arms. There were no significant changes over time or between treatments noted in total cholesterol, high density lipoprotein cholesterol or triglycerides. There were no significant changes in liver enzymes or other safety parameters with time or between treatments. CONCLUSION: Lactobacillus fermentum did not appear to produce a major change in serum lipid fractions, but a small effect cannot be excluded. PMID- 17126769 TI - Assessment of carotid intima-media thickness in subjects with ischemic cerebrovascular events undergoing endarterectomy. AB - BACKGROUND AND AIM: In clinical settings, the degree of lumen stenosis is the parameter used to select patients for carotid surgery. The present study was designed to measure carotid intima-media thickness (IMT), an indicator of atherosclerotic burden, in a sample of consecutive patients with ischemic cerebrovascular events referred for endarterectomy. METHODS AND RESULTS: Carotid endarterectomy specimens from 55 consecutive patients (age 66+/-10 years) admitted to hospital with recent documented atherothrombotic ischemic cerebrovascular events were compared with 24 carotid arteries from people (age 68+/-11 years) who had died from documented causes unrelated to cerebrovascular disease. Measurement of extracranial carotid atherosclerosis was made from three anatomically defined segments, using image-processing software. A total of 426 cross sections was analyzed. Increasing IMT measures were clearly associated with increased risk of an ischemic event. Single maximum IMT values of 2.33 mm (95% CI, 1.69-2.96) for the common carotid, 2.45 mm (95% CI, 1.97-2.93) for the bifurcation, and 2.23 (95% CI, 1.83-2.64) for the internal carotid were associated with a 75% probability of a cerebrovascular ischemic accident. Receiver operator characteristic curve analyses demonstrated that the diagnostic ability of IMT measurements performed at the level of internal carotid artery to separate cases from controls was greater than common carotid artery or bifurcation measurements. CONCLUSIONS: The present pathology study provides data on IMT in patients admitted to hospital for cerebrovascular accidents and referred for carotid endarterectomy. PMID- 17126770 TI - Effects of moderate-intensity exercise training on plasma biomarkers of inflammation and endothelial dysfunction in older patients with type 2 diabetes. AB - BACKGROUND AND AIMS: Some plasma biomarkers of inflammation and endothelial dysfunction have been recently recognized as important cardiovascular risk factors. Currently, there is little information about the effects of aerobic exercise training on these biomarkers in older adults with type 2 diabetes. We have therefore assessed the effects of a twice-weekly moderate, aerobic exercise programme, without a concomitant weight loss diet, on plasma inflammatory and endothelial dysfunction biomarkers in older type 2 diabetic patients. METHODS AND RESULTS: A group of 16 sedentary, overweight, non-smoking, older patients with type 2 diabetes volunteered to participate in a 6-month, supervised, progressive, aerobic training study, two times per week. Plasma levels of hs-C-reactive protein (hs-CRP), soluble tumour necrosis factor (TNF)-alpha receptors, P selectin and intercellular adhesion molecule-1 (ICAM-1) were measured before and after physical training. While hs-CRP and soluble TNF-alpha receptors remained essentially unaffected by physical training, plasma concentrations of P-selectin (P<0.001) and ICAM-1 (P<0.01) markedly decreased; physical training also increased HDL cholesterol by 12% (P<0.05) and decreased uric acid levels by approximately 33% (P=0.021). Body weight, waist circumference, blood pressure, haemoglobin A1c, plasma triglyceride and LDL cholesterol concentrations did not change. Interestingly, the exercise-induced changes in ICAM-1 and P-selectin levels remained significant after adjustment for the percent variations of body weight, waist circumference, haemoglobin A1c, HDL cholesterol and uric acid concentrations. CONCLUSIONS: A twice-weekly, 6-month, progressive aerobic training programme, without a concomitant weight loss diet, is associated with significant decreases in circulating P-selectin and ICAM-1 levels and with a less atherogenic lipid profile in overweight, non-smoking, older type 2 diabetic individuals. PMID- 17126771 TI - Concomitant consumption of red wine and polyunsaturated fatty acids in edible oil does not influence the peroxidation status of chylomicron lipids despite increasing plasma catechin concentration. AB - BACKGROUND AND AIMS: Atherosclerosis may be ameliorated by red wine consumption possibly by providing antioxidants. The effects of red wine on the peroxidation status of chylomicrons (CM) are unknown. The aims were to compare the lipid peroxidation status of oil rich in polyunsaturated fatty acids (PUFA) from a standardised high-fat meal with that of the CM at peak concentrations following ingestion of the meal with and without wine, and to examine the contribution of wine to the antioxidant content of the plasma. METHODS AND RESULTS: Fasted subjects ingested the meal randomly with and without red wine. The peroxidation status was described by conjugated dienes (CD), lipid hydroperoxides (LOOH) and thiobarbituric acid reactive substances (TBARS). CM lag times and the area under the curve (AUC) of CD were determined under oxidative stress. Plasma catechin concentrations and oxygen radical absorbance capacity (ORAC) values were determined. The CM produced with and without wine did not differ in their concentration of CD, LOOH and TBARS. Lag times of CM with and without wine were not significantly different, nor were the AUC. Plasma catechin values increased significantly after consumption of wine with the meal, whereas ORAC values did not. CONCLUSION: Red wine consumption increases plasma catechins, but does not influence lipid peroxidation in postprandial CM. PMID- 17126772 TI - Dietary patterns: a Mediterranean diet score and its relation to clinical and biological markers of cardiovascular disease risk. AB - BACKGROUND AND AIM: It has been suggested that overall dietary patterns and not single nutrients should be studied, since food items might have a synergistic and antagonistic effect on health. The Mediterranean diet has long been associated with lower incidence of cardiovascular disease and cancer. Therefore, we developed a diet score that incorporates the inherent characteristics of this dietary pattern. METHODS AND RESULTS: We used 11 main components of the Mediterranean diet (non-refined cereals, fruits, vegetables, potatoes, legumes, olive oil, fish, red meat, poultry, full fat dairy products and alcohol). For the consumption of items presumed to be close to this pattern we assigned scores 0, 1, 2, 3, 4 and 5 when a participant reported no consumption, rare, frequent, very frequent, weekly and daily, respectively. For the consumption of foods presumed to be away from this pattern we assigned the scores on a reverse scale. Especially for alcohol, we assigned score 5 for consumption of less than 300 ml/day, score 0 for consumption of more than 700 ml/day or none and scores 1-4 for consumption of 300-400, 400-500, 500-600, and 600-700 ml/day (100 ml = 12 g ethanol), respectively. Then a total score ranging from 0 to 55 was calculated. After having applied this diet score in the participants of the ATTICA study we observed a significant positive association with monounsaturated fat and monounsaturated-to-saturated fat intake. We also observed, an inverse association with serum lipids, blood pressures, inflammation and coagulation markers related to cardiovascular disease. The application of that score in a case-control study (CARDIO2000) suggested that the score was inversely associated with the odds of having acute coronary syndromes. CONCLUSION: The Mediterranean diet score proposed above may be useful in assessing the nutritional status of an individual and investigating the relationship of the Mediterranean diet with various health outcomes. PMID- 17126773 TI - Anesthesia awareness: an analysis of its incidence, the risk factors involved, and prevention. PMID- 17126774 TI - Gender differences between predicted and measured propofol C(P50) for loss of consciousness. AB - OBJECTIVE: To investigate gender differences in the effective dose of 50% for loss of consciousness (C(P50LOC)) for propofol using Diprifusor, the most commonly used target-controlled infusion system. DESIGN: Prospective, randomized, comparative study. SETTING: University-affiliated hospital. PATIENTS: 50 ASA physical status I and II patients, aged 20 to 50 years, scheduled for minor surgery. INTERVENTIONS: Patients were randomized into two groups of 25 patients each. A target-controlled infusion of propofol (Diprifusor) was maintained at a predetermined target concentration. After a 10-minute steady state, blinded investigators evaluated patients' consciousness using verbal commands. The propofol test concentration was predetermined using a modified version of Dixon's up-and-down method (starting at 2.5 mug/mL; step size of 0.1 microg/mL). MEASUREMENT: Predicted and measured C(P50LOC) values and bispectral index (BIS) were obtained by averaging the crossover midpoint (ie, consciousness to unconsciousness). Those values were analyzed by unpaired t test: P < 0.05 was considered significant. RESULTS: The predicted C(P50LOC) for men was 2.14 +/- 0.10 microg/mL, which was lower than that for women, 2.55 +/- 0.11 microg/mL (P < 0.0001). No significant difference was found for measured C(P50LOC) in men (2.37 +/- 0.41 microg/mL) and in women (2.30 +/- 0.28 microg/mL) or for BIS measurements. CONCLUSION: Predicted C(P50LOC) by Diprifusor for men tended to be underestimated; that for women tended to be overestimated. Our data support a review of Diprifusor (Astra Zeneca, Osaka, Japan) pharmacokinetic parameters to avoid awareness during operation, particularly for women. PMID- 17126775 TI - Adjuvant bupivacaine scalp block facilitates stabilization of hemodynamics in patients undergoing craniotomy with general anesthesia: a preliminary report. AB - STUDY OBJECTIVE: To evaluate the effect of 0.25% bupivacaine scalp block on alterations in hemodynamics and plasma catecholamine metabolites during general anesthesia in patients undergoing frontotemporal craniotomy. DESIGN: Prospective, clinical study. SETTING: Operating room of a university hospital. PATIENTS: 16 ASA physical status II and III patients who were scheduled for frontotemporal craniotomy. INTERVENTIONS: Patients were prospectively randomized to receive a saline control (C group) or bupivacaine scalp block (SB group) as an adjuvant to general anesthesia using isoflurane in 50% N(2)O-O(2). MEASUREMENTS: Routine monitoring of electrocardiogram, heart rate (HR), and mean arterial blood pressure (MAP) were recorded at two-minute intervals from the beginning of anesthesia until 10 minutes after incision, followed by 5-minute intervals throughout the remaining course of the surgery. By prospective design, increases in MAP or HR by 20% above the mean baseline values were treated with 2.5 mg/kg of thiopental combined with 2 mug/kg of fentanyl. Arterial blood was sampled at 5 minutes before and after skin incision and at the start of dural opening for measuring serum catecholamine metabolites by high-performance liquid chromatography. MAIN RESULTS: Only two patients in the SB group needed additional anesthetics for stabilizing their hemodynamics during the course of anesthesia. In contrast, all C group patients required supplemental anesthesia for controlling the abrupt rise in hemodynamic parameters. In addition, absolute MAP and HR values were significantly higher in the C group than in the SB group during the surgical period between incision and dural opening. The differences in hemodynamics observed between the two groups were, however, not accompanied with a significant change in plasma catecholamine metabolites at each predetermined time interval measured. CONCLUSIONS: Pretreatment with 0.25% bupivacaine scalp block appeared to be an effective adjuvant treatment for maintaining stable hemodynamics for patients undergoing craniotomy during general anesthesia especially at the time of skin incision and dural opening. This study design was unable to discern any correlation between elevation in hemodynamic parameters and a rise in serum catecholamine levels. PMID- 17126776 TI - Blood-sparing efficiency of transdermal nitroglycerine during open fixation of femur shaft fractures: a randomized, double-blind study. AB - STUDY OBJECTIVES: To evaluate the effect of transdermal nitroglycerine for reducing blood loss during open fixation of femur shaft fractures. DESIGN: Randomized, double blind, clinical trial. SETTING: University-affiliated hospital. PATIENTS: 60 ASA physical status I and II patients undergoing elective open fixation of femur shaft fractures. INTERVENTIONS: Patients were randomized to two groups: those who received 3-in (7.5 cm) transdermal nitroglycerine (group N) or 3-in (7.5 cm) placebo (group P). All the drugs used for induction and maintenance of anesthesia were similar in both groups and administered according to the patient's weight. MEASUREMENTS: In both groups, baseline heart rate and blood pressure were recorded and then measured every 5 minutes thereafter. Routine estimation, as well as a mathematical model of blood loss calculation, was used to determine intraoperative blood loss. MAIN RESULTS: Group N had a significant lower systolic, diastolic, and mean arterial blood pressure than those of group P during the operation. The mean estimated blood loss in group N (443.4 +/- 158.0 mL) was significantly lower than that in group P (950.0 +/- 554.9 mL). The calculated blood loss (622.7 +/- 192.4 mL) in group N was also significantly lower than that in group P (1232.1 +/- 630.4 mL). CONCLUSION: By inducing moderate hypotension, the use of transdermal nitroglycerine decreases intraoperative blood loss and the need for transfusion. PMID- 17126777 TI - A survey evaluating the training of anesthesiology residents in office-based anesthesia. AB - STUDY OBJECTIVE: To determine the current status and future plans of residency programs to train residents in office-based anesthesiology (OBA), which is rapidly emerging as an important specialty within the field of anesthesiology. DESIGN: Survey questionnaire. SETTING: University hospital. MEASUREMENTS: A 14 question survey was sent to the 134 listed members of the Society of Academic Anesthesia Chairs/Association of Anesthesiology Program Directors to elicit the current status of educational endeavors and clinical exposure of anesthesiology residents to this type of practice. MAIN RESULTS: There were 95 respondents (72%). Fifteen (15.8%) academic anesthesiology programs provide OBA services in the community and two (2.1%) of the academic programs provide clinical exposure to their residents. CONCLUSIONS: Residents are receiving minimal, if any, exposure to OBA during their training. PMID- 17126778 TI - Patient perception of the role of anesthesiologists: a perspective from the Caribbean. AB - STUDY OBJECTIVE: To assess patients' perception of the role of an anesthesiologist in a Caribbean country. DESIGN: Self-administered structured questionnaire evaluation. SETTING: Preoperative waiting rooms of three tertiary care teaching hospitals: Port of Spain General Hospital, Eric Williams Medical Sciences Complex, and San Fernando General Hospital, Trinidad. PATIENTS: 424 adult surgical patients awaiting elective surgery. INTERVENTIONS: None. MEASUREMENTS: A questionnaire was devised to test the knowledge of the respondents regarding the job description, attitudes, and various roles of anesthesiologists in the hospital. MAIN RESULTS: 371 completed responses were obtained for analysis. One tenth of the respondents did not know who an anesthesiologist was and 59% of them knew that an anesthesiologist was a doctor; there was a statistically significant association of the educational level of the respondent and this response. Of the respondents, 70% felt that the anesthesiologists were easy to talk to and pleasant by the bedside; 46% responded that the anesthesiologists did not discuss the complications and side effects of drugs before the procedure; 5% considered the anesthesiologists as more important than the surgeon, and 59% considered both equally important. Only 19% responded that they knew that the anesthesiologists had a role in the intensive care unit. CONCLUSIONS: Patients still have inadequate knowledge regarding anesthesiologists and their different roles in hospitals. PMID- 17126779 TI - Concurrent expansion of plasma volume and left ventricular end-diastolic volume in patients after rapid infusion of 5% albumin and lactated Ringer's solution. AB - STUDY OBJECTIVE: To examine the effects of plasma volume expansion on plasma volume, left ventricular end-diastolic volume (LVEDV), and cardiac index (CI) after rapid fluid infusion, as knowledge of the degree of concordance between plasma and cardiac preload expansion could optimize LVEDV expansion without administering excessive fluid. DESIGN: Randomized, double-blinded study. SETTING: Academic community hospital. PATIENTS: 20 patients undergoing elective coronary artery bypass surgery. INTERVENTIONS: Patients were administered either 5% albumin (5 mL/kg) or lactated Ringer's solution (25 mL/kg) over 30 minutes, just before incision. MEASUREMENTS: Serial measurements of plasma volume, LVEDV by transesophageal echocardiography, and CI were recorded. MAIN RESULTS: Albumin expanded plasma volume and LVEDV to a similar degree (11.3% and 13.2%). In contrast, lactated Ringer's solution increased plasma volume more than LVEDV (21.7% vs 14.4%; P = 0.0005). Increased LVEDV significantly but poorly correlated with increased CI (r(2) = 0.2, P < 0.0001) for both fluids. However, LVEDV expansion was brief and returned to baseline or less within 30 minutes for both fluids despite continued plasma volume expansion and increased CI. Correspondingly, rates of decline from peak expansion were significantly faster for LVEDV than plasma volume expansion for both albumin (-1.9% + 1.9%/min vs 0.1% + 0.1%/min; P = 0.0008) and lactated Ringer's (-1.1% + 0.8%/min vs -0.4% + 0.2%/min; P = 0.006). CONCLUSIONS: Intravenous fluids increased LVEDV to a lesser extent and duration than did plasma volume expansion. Monitoring of LVEDV was a poor guide for fluid administration to maximize CI. PMID- 17126780 TI - Effect of postoperative epidural analgesia on morbidity and mortality after lung resection in Medicare patients. AB - STUDY OBJECTIVE: To perform an analysis of the Medicare claims database in patients undergoing lung resection to determine whether there is an association between postoperative epidural analgesia and mortality. DESIGN: Retrospective cohort (database) design. SETTING: University hospital. MEASUREMENTS: We examined a cohort of 3501 patients obtained from a 5% nationally random sample of 1997 to 2001 Medicare beneficiaries who underwent nonemergency segmental excision of the lung (International Classification of Diseases, 9th Revision, Clinical Modification codes 32.3 and 32.4). Patient data were divided into two groups depending on the presence or absence of billing for postoperative epidural analgesia (Current Procedural Terminology code 01996). The primary outcomes assessed were death at 7 and 30 days after the procedure. The rates of major morbidity (acute myocardial infarction, angina, cardiac dysrhythmias, heart failure, pneumonia, pulmonary edema, respiratory failure, deep venous thrombosis, pulmonary embolism, sepsis, acute renal failure, somnolence, acute cerebrovascular event, transient organic syndrome, and paralytic ileus) were also compared. Multivariate regression analysis incorporating race, gender, age, comorbidities, hospital size, hospital teaching status, and hospital technology status was performed to determine whether the presence of postoperative epidural analgesia had an independent effect on mortality or major morbidity. MAIN RESULTS: Multivariate regression analysis showed that the presence of epidural analgesia was associated with a significantly lower odds of death at 7 days (odds ratio, 0.39; 95% confidence interval, 0.19-0.80; P = 0.001) and 30 days (odds ratio, 0.53; 95% confidence interval, 0.35-0.78; P = 0.002) after surgery. There was no difference between the groups with regard to overall major morbidity. CONCLUSIONS: Postoperative epidural analgesia may contribute to lower odds of death after segmental excision of the lung, although the mechanism of such a benefit is not clear from our analysis. PMID- 17126781 TI - Comparison of ropivacaine and bupivacaine for intrathecal anesthesia during outpatient arthroscopic surgery. AB - STUDY OBJECTIVE: To compare the effects of intrathecal ropivacaine with bupivacaine in a dose ratio of 2:1 for outpatient arthroscopic knee surgery. DESIGN: Randomized, single-blinded study. SETTING: University-affiliated hospital. PATIENTS: 90 patients scheduled for outpatient arthroscopic knee surgery. INTERVENTIONS: Patients were randomized and assigned in single-blinded fashion to receive a 3-mL solution of either 15 mg of isobaric ropivacaine (group R; n = 45) or 7.5 mg of isobaric bupivacaine (group B; n = 45) through a 27-gauge Quincke spinal needle at the L(3) to L(4) interspace, while placed in the lateral decubitus position. MEASUREMENTS: Onset and offset times for sensory and motor block; highest level of sensory block; duration of the sensory and motor block; first ambulation, urination, and discharge time; mean arterial pressure; and heart rate were all recorded. MAIN RESULTS: Onset time for sensory block (mean +/ SD) to L1 and time until sensory block regressed to L2 were shorter in group R. Complete motor block occurred in 40 patients with ropivacaine and 45 patients with bupivacaine. First ambulation and first urination and discharge times were similar between the two groups. Cephalad spread of sensory block was higher with ropivacaine (P < 0.05). The median (range) upper sensory level obtained with bupivacaine was T11 (T6-L1) and T8 (T6-T10) with ropivacaine. Hemodynamic changes were similar between the groups (P > 0.05). CONCLUSION: Isobaric ropivacaine 15 mg provided a higher sensory block level and shorter sensorial onset and offset times than did 7.5 mg of isobaric bupivacaine. PMID- 17126782 TI - Influence of aging on lidocaine requirements for pain on injection of propofol. AB - STUDY OBJECTIVE: To evaluate the influence of aging on lidocaine requirements for propofol-induced pain on injection. DESIGN: Prospective, randomized, double blind, placebo-controlled study. SETTING: The study was undertaken at a University hospital. PATIENTS: 160 ASA physical status I and II adult patients scheduled for elective surgery with general anesthesia. INTERVENTIONS: Patients received placebo (saline) or lidocaine intravenously at three different doses (10, 20, or 40 mg), with venous occlusion for two minutes, followed by injection of propofol 0.5 mg/kg into a dorsal hand vein. MEASUREMENT AND MAIN RESULTS: Pain during injection of propofol was evaluated. For young patients, the frequency of propofol-induced pain was 70% in patients receiving lidocaine 10 mg (P = not significant); 50% in those receiving lidocaine 20 mg (P = not significant); and 30% in those receiving lidocaine 40 mg (P < 0.05), compared with placebo (80%). For elderly patients, 15 patients (75%) complained of pain in the placebo group, compared with 13 (65%) in the lidocaine 10 mg group (P = not significant); 5 (25%) in the lidocaine 20 mg group (P < 0.05); and 4 (20%) in the lidocaine 40 mg group (P < 0.05). CONCLUSIONS: A lidocaine dose of 40 mg for young patients and 20 mg for old patients, with venous occlusion for two minutes, is sufficient to reduce pain on injection of propofol. PMID- 17126783 TI - Recurrent intraoperative coronary artery spasm in a patient with coronary artery aneurysm. AB - Coronary artery spasm (CAS) may result from endothelial dysfunction in nonobstructed arteries. Its complications include myocardial infarction, dysrhythmias, and death. It is difficult to diagnose CAS during general anesthesia because it closely mimics obstructive ischemia. We report the case of a patient who demonstrated intraoperative CAS during two separate surgeries. Angiography after the first surgery showed nonobstructed coronary anatomy. During the second surgery, recurrence of acute ST elevations caused by CAS rapidly responded to nitroglycerin. A high index of suspicion for CAS as a cause of intraoperative ischemia is necessary for an early diagnosis, treatment, and a favorable outcome. PMID- 17126784 TI - Bedside ultrasonography in the differential diagnosis of pulmonary pathologies in the intensive care unit. AB - Among critically ill patients, opacification of a part or whole lung field on chest radiography may pose a challenge in the differential diagnosis of acute pulmonary pathologies (eg, pneumothorax, hemothorax, pleural effusion, atelectasis, and solid organ in thoracic cavity) and selection of treatment modalities. In cases in which clinical findings, history, and imaging studies are not conclusive, bedside ultrasonography may be invaluable in achieving a diagnosis. We present two cases in which portable ultrasonography at the bedside was critical to the diagnosis and subsequent management of the patient. PMID- 17126785 TI - Acute coronary syndrome and myocardial infarction after orthopedic surgery in a patient with a recently placed drug-eluting stent. AB - Providing anesthesia care for patients who have recently undergone intracoronary drug-eluting stent placement presents unique clinical challenges. It is currently recommended that these patients remain on antiplatelet therapy until reendothelialization of the vessel has occurred (ie, 3-6 months, depending on the eluting medication) to prevent stent restenosis. In the setting of urgent or emergent surgery, it may not be possible to wait until a full course of antiplatelet therapy has been completed. We report an unusual case of postoperative acute coronary syndrome in a gentleman who underwent intracoronary stenting 7 weeks before nonelective revision hip arthroplasty. To our knowledge, this is the first case in the anesthesia literature to report postoperative cardiac morbidity after recent drug-eluting stent deployment. PMID- 17126786 TI - Variant location of the musculocutaneous nerve during axillary nerve block. AB - We present the case of a 56-year-old man who underwent axillary nerve block for a wrist arthroscopy procedure, with real-time ultrasound and peripheral nerve stimulator guidance. The ulnar nerve and radial nerve were located medial and posterior to the brachial artery, respectively. A large complex structure was noted in the position typically occupied by the median nerve. Contact of this structure with the stimulating needle produced strong biceps contraction, and slight adjustment of the needle resulted in forearm pronation. After injection of 10 mL of local anesthetic near this structure, it appeared to consist of two separate components on ultrasound. We believe that these components represented the median and musculocutaneous nerves lying together, lateral to the artery. Radial, median, ulnar, and musculocutaneous nerve block ensued, and wrist arthroscopy was carried out uneventfully. Knowledge of this anatomical variation may improve anesthesiologists' ability to provide effective axillary block. PMID- 17126787 TI - The use of recombinant factor VIIa (NovoSeven) for treatment of active or impending bleeding in brain injury: broadening the indications. AB - We report three patients with severe traumatic brain injury, both open and closed, who were treated with recombinant activated factor VII. This treatment was given in a desperate, last-ditch effort to save the life of patient 1, as a preventive or early treatment of a developing hematoma in patient 2, and as treatment of a threatening hematoma in patient 3. One of the three patients survived. During the past few years we have broadened the indications for recombinant activated factor VII and started using it as a preventive measure rather than as a "last line of defense." However, the potential complications of disseminated intravascular coagulation and thrombotic events, as well as the cost effectiveness in view of the available evidence-based medicine, should be considered. PMID- 17126788 TI - Workforce economics in anesthesia: the tipping point. PMID- 17126789 TI - Support methods for a popliteal approach to the sciatic nerve block. PMID- 17126790 TI - Suspected preoperative takotsubo cardiomyopathy. PMID- 17126791 TI - Another knotted epidural catheter. PMID- 17126792 TI - Potential aspiration hazard when using a nasopharyngeal temperature probe with disposable cover. PMID- 17126793 TI - Gastric aspiration in patients with a laryngectomy stoma: is it possible? PMID- 17126794 TI - Dynamics of signals and structures. PMID- 17126795 TI - [The PAI-1 swing: microenvironment and cancer cell migration]. AB - Cancer is a complex and dynamic process caused by a cellular dysfunction leading to a whole organ or even organism vital perturbation. To better understand this process, we need to study each one of the levels involved, which allows the scale change, and to integrate this knowledge. A matricellular protein, PAI-1, is able to induce in vitro cell behaviour modifications, morphological changes, and to promote cell migration. PAI-1 influences the mesenchymo-amaeboid transition. This matricellular protein should be considered as a potential 'launcher' of the metastatic process acting at the molecular, cellular, tissular levels and, as a consequence, at the organism's level. PMID- 17126796 TI - Cooperation strategies, signals and symbiosis. AB - The authors of models concerning cooperation found an interesting application point in mutualism and symbiosis. Phenomena which are to be found in symbiosis are integrated by the various models in relation to the 'Prisoner's Dilemma': 'cost-benefit' ratio components, signals, neighbourhood interaction or reciprocity. Reciprocal altruism is found in symbiosis, but others strategies seem more to reflect human societies, where choice is unrestricted. It is very difficult to give a value to cost and benefit equilibriums. The biological environment interferes and a model based on a dominant strategy is often limited or insufficient to report on the living system's evolution. PMID- 17126797 TI - Modelling molecular interactions with game networks' theory. AB - We present a method to model biological systems, the theory of games networks. It extends game theory by multiplying the number of games, and by allowing agents to play several games simultaneously. Some important notions of biological systems, such as locality of interactions and modularity, can then be modelled. PMID- 17126798 TI - Model of interactions in biology and application to heterogeneous network in yeast. AB - A major challenge for bioinformatics and theoretical biology is to build and analyse a unified model of biological knowledge resulting from high-throughput experiment data. Former work analyzed heterogeneous data (protein-protein interactions, genetic regulation, metabolism, synexpression) by modelling them by graphs. These models are unable to represent the qualitative dynamics of the reactions or to model the n-ary interactions. Here, MIB, the Model of Interactions in Biology, a bipartite model of biological networks, is introduced, and its use for topological analysis of the heterogeneous network is presented. Heterogeneous loops and links between synexpression pattern and underlying molecular mechanisms are proposed. PMID- 17126799 TI - Potential automata. Application to the genetic code III. AB - In previous notes, we have described both mathematical properties of potential (n switches) and potential-Hamiltonian (Lienard systems) continuous differential systems, and also biological applications, especially those concerning primitive cyclic RNAs related to the genetic code. In the present note, we give a general definition of a potential automaton, and we show that a discrete Hopfield-like system already introduced by Goles et al. is a good candidate for such a potential automaton: it has a Lyapunov functional that decreases on its trajectories and whose time derivative is just its discrete velocity. Then we apply this new notion of potential automaton to the genetic code. We show in particular that the consideration of only physicochemical properties of amino acids, like their molecular weight, hydrophobicity and ability to create hydrogen bonds suffices to build a potential decreasing on trajectories corresponding to the synonymy classes of the genetic code. Such an 'a minima' construction reinforces the classical stereochemical hypothesis about the origin of the genetic code and authorizes new views about the optimality of its synonymy classes. PMID- 17126800 TI - Steady-state kinetic behaviour of two- or n-enzyme systems made of free sequential enzymes involved in a metabolic pathway. AB - The overall rate of functioning of a set of free sequential enzymes of the Michaelis-Menten type involved in a metabolic pathway has been computed as a function of the concentration of the initial substrate under steady-state conditions. Curves monotonically increasing up to a saturation plateau have been obtained in all cases. The shape of these curves is sometimes, but not usually, close to that of a hyperbola. Cases exist in which the overall rate of reaction becomes quasi proportional to the concentration of initial substrate almost up to the saturation plateau, which never occurs with individual enzymes. Increasing the number of enzymes sequentially involved in a metabolic pathway does not seem to generate any particularly original behaviour compared with that of two-enzyme systems. PMID- 17126801 TI - Approximation for limit cycles and their isochrons. AB - Local analysis of trajectories of dynamical systems near an attractive periodic orbit displays the notion of asymptotic phase and isochrons. These notions are quite useful in applications to biosciences. In this note, we give an expression for the first approximation of equations of isochrons in the setting of perturbations of polynomial Hamiltonian systems. This method can be generalized to perturbations of systems that have a polynomial integral factor (like the Lotka-Volterra equation). PMID- 17126802 TI - Storage and recall of environmental signals in a plant: modelling by use of a differential (continuous) formulation. AB - The breaking of the symmetry of bud growth in Bidens seedlings involves a sort of plant 'memory'. An asymmetrical stimulus (e.g., the pricking of one of the seedling cotyledons) stores a 'symmetry-breaking' signal within the plants (function STO). Depending on other stimuli received by the seedlings, the stored signal may remain silent or be recalled (RCL function) and take effect in the seedling morphogenesis (asymmetry of the growth of the cotyledonary buds, with a statistical advantage to the bud at the axil of the non-stimulated cotyledon). We show that this memory mechanism can be interpreted by a model taking into account a genetic control exerted on a non-linear enzymatic system that is able to choose trajectories going to different attractors, depending on the stimulation intensity. PMID- 17126804 TI - A high-throughput method for fitting dose-response curves using Microsoft Excel. PMID- 17126803 TI - Novel inhibitors of the trypanosome alternative oxidase inhibit Trypanosoma brucei brucei growth and respiration. AB - African trypanosomiasis is a deadly disease for which few chemotherapeutic options are available. The causative agents, Trypanosoma brucei rhodesiense and T. b. gambiense, utilize a non-cytochrome, alternative oxidase (AOX) for their cellular respiration. The absence of this enzyme in mammalian cells makes it a logical target for therapeutic agents. We designed three novel compounds, ACB41, ACD15, and ACD16, and investigated their effects on trypanosome alternative oxidase (TAO) enzymatic activity, parasite respiration, and parasite growth in vitro. All three compounds contain a 2-hydroxybenzoic acid moiety, analogous to that present in SHAM, and a prenyl side chain similar to that found in ubiquinol. ACD15 and ACD16 are further differentiated by the presence of a solubility enhancing carbohydrate moiety. Kinetic studies with purified TAO show that all three compounds competitively inhibit TAO, and two compounds, ACB41 and ACD15, have inhibition constants five- and three-fold more potent than SHAM, respectively. All three compounds inhibited the respiration and growth of continuously cultured T. b. brucei bloodstream cells in a dose-dependent manner. None of the compounds interfered with respiration of rat liver mitochondria, nor did they inhibit the growth of a continuously cultured mammalian cell line. Collectively, the results suggest we have identified a new class of compounds that are inhibitors of TAO, have trypanocidal properties in vitro, and warrant further investigation in vivo. PMID- 17126806 TI - Protective effect of the in situ pellicle on dentin erosion - an ex vivo pilot study. AB - AIM: The acquired pellicle is well known as an anti-erosive proteinaceous layer on enamel, but its protective properties on dentin have not been investigated in detail until now. The aim of the present ex vivo study was to evaluate the erosive effects on pellicle coated dentin. METHODS: Bovine dentin slabs were exposed to the oral cavity of one subject for 120 min for in situ pellicle formation. Subsequently, the slabs were incubated with HCl (pH 2.3) in vitro for 5 min and erosive calcium-release was measured photometrically. In addition, the acid treated specimens were evaluated by transmission electron microscopy (TEM). Pellicle free samples served as controls. RESULTS: Calcium erosion from the pellicle coated dentin slabs amounted to 23.5+/-2.9 microg Ca/min (pellicle free samples: 32.2+/-4.2 microg Ca/min). The difference was statistically significant (p < or = 0.05). In pellicle coated as well as in uncoated dentin samples, TEM evaluation showed a demineralised dentinal surface layer which thickness ranged between 3 and 6 microm. The pellicle itself was partially dissolved but not removed by hydrochloric acid treatment. CONCLUSION: The protective properties of the acquired pellicle against an erosive challenge of the dentinal surface are limited. The dentinal pellicle functions like an ion permeable network rather than a barrier. PMID- 17126805 TI - Targeted expression of csCSF-1 in op/op mice ameliorates tooth defects. AB - OBJECTIVE: The aim of this study was to characterize the tooth phenotype of CSF-1 deficient op/op mice and determine whether expression of csCSF-1 in these mice has a role in primary tooth matrix formation. DESIGN: Ameloblasts and odontoblasts, isolated from wt/wt frozen sections using laser capture microdissection, were analysed for csCSF-1, sCSF-1 and CSF-1R mRNA by RT-PCR. Mandibles, excised from 8 days op/op and wt/wt littermates, were examined for tooth morphology as well as amelogenin and DMP1 expression using in situ hybridisation. op/opCS transgenic mice, expressing csCSF-1 in teeth and bone using the osteocalcin promoter, were generated. Skeletal X-rays and histomorphometry were performed; teeth were analysed for morphology and matrix proteins. RESULTS: Normal dental cells in vivo express both CSF-1 isoforms and CSF-1R. Compared to wt/wt, op/op teeth prior to eruption showed altered dental cell morphology and dramatic reduction in DMP1 transcripts. op/opCS mice showed marked resolution of osteopetrosis, tooth eruption and teeth that resembled amelogenesis imperfecta-like phenotype. At 3 weeks, op/op teeth showed severe enamel and dentin defects and barely detectable amelogenin and DMP1. In op/opCS mice, DMP1 in odontoblasts increased to near normal and dentin morphology was restored; amelogenin also increased. Enamel integrity improved in op/opCS, although it was thinner than wt enamel. CONCLUSIONS: Results demonstrate that ameloblasts and odontoblasts are a source and potential target of CSF-1 isoforms in vivo. Expression of csCSF-1 within the tooth microenvironment is essential for normal tooth morphogenesis and may provide a mechanism for coordinating the process of tooth eruption with endogenous matrix formation. PMID- 17126807 TI - Osteoclast formation is strongly reduced both in vivo and in vitro in the absence of CD47/SIRPalpha-interaction. AB - Physical interaction between the cell surface receptors CD47 and signal regulatory protein alpha (SIRPalpha) was reported to regulate cell migration, phagocytosis, cytokine production, and macrophage fusion. However, it is unclear if the CD47/SIRPalpha-interaction can also regulate macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor (NF)-kappaB ligand (RANKL)-stimulated formation of osteoclasts. Here, we show that functional blocking antibodies to either CD47 or SIRPalpha strongly reduced formation of multinucleated tartrate-resistant acid phosphatase (TRAP)+ osteoclasts in cultures of murine hematopoietic cells, stimulated in vitro by M-CSF and RANKL. In addition, the numbers of osteoclasts formed in M-CSF/RANKL-stimulated bone marrow macrophage cultures from CD47-/- mice were strongly reduced, and bones of CD47-/- mice exhibited significantly reduced osteoclast numbers, as compared with wild-type controls. We conclude that the CD47/SIRPalpha interaction is important for M-CSF/RANKL-stimulated osteoclast formation both in vivo and in vitro, and that absence of CD47 results in decreased numbers of osteoclasts in CD47-/- mice. PMID- 17126808 TI - Developmental stage- and DNA damage-specific functions of C. elegans FANCD2. AB - In this study, we set out to investigate the role of Fanconi anemia complementation group D2 protein (FANCD2) in developmental stage-specific DNA damage responses in Caenorhabditis elegans. A mutant C. elegans strain containing a deletion in the gene encoding the FANCD2 homolog, FCD-2, exhibited egg-laying defects, precocious oogenesis, and partial defects in fertilization. The mutant strain also had a lower hatching rate than the wild-type after gamma-irradiation of embryos, but not after the irradiation of pachytene stage germ cells. This mutation sensitized pachytene stage germ cells to the genotoxic effects of photoactivated psoralen, as seen by a greatly reduced hatching rate and increased chromosomal aberrations. This mutation also enhanced physiological M-phase arrest and apoptosis. Taken together, our data reveal that the C. elegans FANCD2 homolog participates in the repair of spontaneous DNA damage and DNA crosslinks, not only in proliferating cells but also in pachytene stage cells, and it may have an additional role in double-stranded DNA break repair during embryogenesis. PMID- 17126809 TI - A novel model to identify interaction partners of the PTEN tumor suppressor gene in human bladder cancer. AB - Phosphatase and tensin homolog (PTEN), deleted on chromosome 10, is a potent tumor suppressor. PTEN expression is reduced in advanced bladder cancer and reduction correlates with disease stage. To gain insights into the function of PTEN in human bladder cancer by identifying its binding partners, we developed a novel IPTG inducible PTEN expression system and evaluated this system in the PTEN null UMUC-3 human bladder cancer xenograft model. In this model, induction of PTEN in vivo resulted in reduced tumor growth. We used mass spectrometry to identify PTEN interaction partners in these cells, which identified known interaction partners major vault protein (MVP) and paxillin as well as a novel interaction partner, TRK fused gene (TFG). In conclusion, using a biologically relevant model system to dissect PTEN tumor suppressor function in human bladder cancer, we identified three molecules important for many cellular functions in complex with PTEN. PMID- 17126810 TI - Conserved motifs in voltage-sensing and pore-forming modules of voltage-gated ion channel proteins. AB - Voltage-gated ion channels (VGCs) mediate selective diffusion of ions across cell membranes to enable many vital cellular processes. Three-dimensional structure data are lacking for VGC proteins; hence, to better understand their function, there is a need to identify the conserved motifs using sequence analysis methods. In this study, we have used a profile-to-profile alignment method to identify several new conserved motifs specific to each transmembrane segment (TMS) of the voltage-sensing and the pore-forming modules of Ca2+, Na+, and K+ channel subfamilies. For Ca2+ and Na+, the functional theme of motif conservation is similar in all segments while they differ with those of the K+ channel proteins. Nevertheless, the conservation is strikingly similar in the S4 segment of the voltage-sensing module across all subfamilies. In each subfamily and for each TMS, we have identified conserved motifs/residues and correlated their functional significance and disease associations in human, using mutational data from the literature. PMID- 17126811 TI - Hypoxia/reoxygenation-induced cytotoxicity in cultured human lymphocytes. AB - Reactive oxygen species (ROS) generated after exposure to hypoxia and reoxygenation (H/R) play a pivotal role in the stimulation of cell death. In this study, we explored H/R-induced cytotoxicity in human lymphocytes. Compared to cells under normoxic conditions, H/R-treated cells exhibited significantly decreased viability and increased DNA breakage. Western blotting analysis demonstrated that H/R-induced the accumulation of p53 and p63 proteins. H/R also led to the activation of caspase-3 and -9, accompanied by the cleavage of PARP (poly(ADP-ribose)polymerase). Because apoptosis is usually accompanied by ROS generation and collapse of the mitochondrial membrane potential (MMP, Deltapsi(m)), we examined ROS and MMP levels in H/R-treated lymphocytes. Cells subjected to H/R exhibited significantly increased ROS and decreased MMP, compared with normoxic cells. Taken together, these results indicate that H/R treatment of human lymphocytes induces rapid ROS generation and MMP collapse, which triggers apoptosis. PMID- 17126812 TI - Ablation of the mammalian methionine sulfoxide reductase A affects the expression level of cysteine deoxygenase. AB - Methionine sulfoxide reductases (Msrs) are able to reduce methionine sulfoxide to methionine both in proteins and free amino acids. By their action it is possible to regulate the function of specific proteins and the cellular antioxidant defense against oxidative damage. Similarly, cysteine deoxygenase (CDO) may be involved in the regulation of protein function and antioxidant defense mechanisms by its ability to oxidized cysteine residues. The two enzymes' involvement in sulfur amino-acids metabolism seems to be connected. Lack of methionine sulfoxide reductase A (MsrA) in liver of MsrA-/- led to a significant drop in the cellular level of thiol groups and lowered the CDO level of expression. Moreover, following selenium deficient diet (applied to decrease the expression levels of selenoproteins like MsrB), the latter effect was maintained while the basal levels of thiol decreased in both mouse strains. We suggest that both enzymes are working in coordination to balance cellular antioxidant defense. PMID- 17126813 TI - Interaction between the SH3 domains and C-terminal proline-rich region in NADPH oxidase organizer 1 (Noxo1). AB - NADPH oxidase organizer 1 (Noxo1), harboring a PX domain, two SH3 domains, and a proline-rich region (PRR), participates in activation of superoxide-producing Nox family NADPH oxidases. Here, we show that Noxo1 supports superoxide production in a cell-free system for gp91(phox)/Nox2 activation by arachidonic acid. This lipid enhances an SH3-mediated binding of Noxo1 to p22(phox), a protein complexed with Nox oxidases; the binding is known to be required for Nox activation. We also demonstrate that the bis-SH3 domain directly interacts with the Noxo1 PRR. The interaction appears to prevent the bis-SH3 domain and PRR from binding to their target proteins; disruption of the intramolecular interaction facilitates Noxo1 binding to p22(phox) and also allows the PRR to associate with the Nox activator Noxa1, which association is crucial for Nox activation as well. These findings suggest that Nox activation involves a conformational change leading to disruption of the bis-SH3-PRR interaction in Noxo1. PMID- 17126814 TI - Metal chelators react also with reactive oxygen and nitrogen species. AB - Popular chelators (desferrioxamine, SIH, EDTA, EGTA, DTPA, and NTA) were demonstrated to have antioxidant properties, being able to reduce ABTS radical cation and react with peroxyl radicals, peroxynitrite, and hypochlorite. Desferrioxamine and SIH were most potent antioxidants in all cases. These results point to the necessity of a careful interpretation of experiments in which the inhibition of free radical reactions by antioxidants is used as a proof of involvement of metal ions in a reaction. PMID- 17126815 TI - Allelic variants of ABC drug transporter Cdr1p in clinical isolates of Candida albicans. AB - Candida drug resistance protein (Cdr1p) is a major drug efflux protein, which plays a key role in commonly encountered clinical azole resistance in Candida albicans. We have analyzed its sequence in several azole resistant clinical isolates to evaluate the allelic variation within CDR1 gene and to relate it to its functional activity. The sequence analysis revealed 53 single nucleotide polymorphisms (SNPs), out of which six were non-synonymous single nucleotide polymorphisms (NS-SNPs) implying a change in amino acid and were found in two or more than two allelic combinations in different sensitive or resistant isolates. We have identified three new NS-SNPs namely, E948P, T950S, and F1399Y, in isolates wherein F1399Y appeared to be unique and was present in one of the naturally occurring azole resistant isolates obtained from Indian diabetic patients. However, site-directed mutagenesis showed that the residue F1399 in between TMS 11 and TMS 12 does not affect the functionality of Cdr1p. Taken together, our SNPs analyses reveal that unlike human P-gp, the naturally acquired allelic variations are mostly present in non-conserved regions of the protein which do not allow Cdr1p to genetically evolve in a manner, that would allow a change in its functionality to affect substrate recognition, specificity, and drug efflux activity of C. albicans cells. PMID- 17126816 TI - Fluorescent protein pair emit intracellular FRET signal suitable for FACS screening. AB - The fluorescent proteins ECFP and HcRed were shown to give an easily resolved FRET-signal when expressed as a fusion inside mammalian cells. HeLa-tat cells expressing ECFP, pHcRed, or the fusion protein pHcRed-ECFP were analyzed by flow cytometry after excitation of ECFP. Cells expressing HcRed-ECFP, or ECFP and HcRed, were mixed and FACS-sorted for FRET positive cells: HcRed-ECFP cells were greatly enriched (72 times). Next, cloned human antibodies were fused with ECFP and expressed anchored to the ER membrane. Their cognate antigens (HIV-1 gp120 or gp41) were fused to HcRed and co-expressed in the ER. An increase of 13.5+/-1.5% (mean+/-SEM) and 8.0+/-0.7% in ECFP fluorescence for the specific antibodies reacting with gp120 or gp41, respectively, was noted after photobleaching. A positive control (HcRed-ECFP) gave a 14.8+/-2.6% increase. Surprisingly, the unspecific antibody (anti-TT) showed 12.1+/-1.1% increase, possibly because overexpression in the limited ER compartment gave false FRET signals. PMID- 17126817 TI - Inducible overexpression of c-Jun in MCF7 cells causes resistance to vinblastine via inhibition of drug-induced apoptosis and senescence at a step subsequent to mitotic arrest. AB - c-Jun is a major component of the AP-1 transcription factor and plays a key role in regulation of diverse biological processes including proliferation and apoptosis. Treatment of a wide variety of cells with the microtubule inhibitor vinblastine leads to a robust increase in c-Jun expression, JNK-mediated c-Jun phosphorylation, and activation of AP-1-dependent transcription. However, the role of c-Jun induction in the response of cells to vinblastine remains obscure. In this study we used MCF7 breast cancer cell lines that express the dominant negative form of c-Jun, TAM-67, as well as cells that overexpress c-Jun, under the control of an inducible promoter. Vinblastine induced c-Jun protein expression, c-Jun phosphorylation, and AP-1 activation in MCF7 cells, and these parameters were strongly inhibited by inducible TAM-67 expression and strongly enhanced by inducible c-Jun expression. Vinblastine-induced cell death was not affected by TAM-67 expression whereas cells were protected by c-Jun overexpression. Further investigation revealed that apoptotic and senescent cells were observed after vinblastine treatment and that both outcomes were strongly inhibited by c-Jun overexpression. Although c-Jun expression inhibited cell death, it did not affect the ability of vinblastine to induce mitotic arrest. These results indicate that c-Jun expression plays a protective role in the cellular response to vinblastine and operates post-mitotic block to inhibit drug induced apoptosis and senescence. PMID- 17126819 TI - Involvement of alcohol and aldehyde dehydrogenase activities on hepatic retinoid metabolism and its possible participation in the progression of rat liver regeneration. AB - Liver alcohol dehydrogenase (ADH) activity is decreased towards exogenous substrates after partial hepatectomy (PH), probably due to putative endogenous substrates acting as ADH inhibitors. Hence, retinoids could be suitable candidates as such endogenous substrates. Therefore, cytosolic ADH kinetic analysis using several substrates, liver cytosolic and mitochondrial aldehyde dehydrogenase (ALDH) activities, retinal and retinol content, as well as expression of proteins for ADH and CRBPI (a retinol carrier protein) were determined in liver samples, at two stages of liver regeneration (one- or two thirds PH). The effect of inhibiting in vivo liver ADH by 4-methylpyrazole (4-MP) was also evaluated after 70%-PH. With 70%-PH, in vitro ADH activity towards exogenous alcohols and aldehydes was diminished, but retinol oxidation was increased and retinal reduction was decreased. These activities that be due to the participation of an ADH type which did not correlate with the amount of immunoreactive ADH protein. Cytosolic and mitochondrial ALDH activities oxidized actively retinal, whereas retinol and CBRP-I expression were reduced in these animals. With 30%-PH, these changes were less evident and sometimes opposite to those found with 70%-PH. In addition, retinol readily inhibited ADH-mediated ethanol oxidation. Interestingly, in vivo 4-MP administration inhibited ADH activity in a dose-dependent manner correlating with a progressive inhibition of liver regeneration. In conclusion, PH-induced inhibition of ADH (mainly type I) seems to be related to ADH-mediated retinoid metabolism during liver proliferation. Thus, results suggest a role of ADH in retinoid metabolism, which is apparently required during rat liver regeneration. PMID- 17126818 TI - Peripheral-type benzodiazepine receptor overexpression and knockdown in human breast cancer cells indicate its prominent role in tumor cell proliferation. AB - The peripheral-type benzodiazepine receptor (PBR), an 18-kDa high affinity drug and cholesterol binding protein, is expressed at high levels in various cancers. Its expression is positively correlated with aggressive metastatic behavior in human breast cancer cells. To determine the role of PBR in tumor progression, two human mammary carcinoma cell lines were utilized: the non-aggressive MCF-7 cell line, which expresses extremely low PBR levels, and the highly aggressive MDA-MB 231 cell line, which has much higher PBR levels. We have generated stably transfected lines of the tetracycline-repressible MCF-7 cell line (MCF-7 Tet-Off) with inducible human PBR cDNA. Induction of PBR expression in MCF-7 Tet-Off cells increased PBR ligand binding and cell proliferation. Transfection of MDA-MB-231 cells with multiple siRNAs complementary to PBR (PBR-siRNAs) led to different levels of PBR mRNA knockdown. Lentiviral-mediated PBR RNA interference in MDA-MB 231 cells decreased PBR levels by 50%. Decreased PBR expression was associated with cell cycle arrest at G2 phase, decreased cell proliferation, and significant increases in the protein levels of the cyclin-dependent kinase inhibitor p21(WAF/CIP1). These changes were accompanied by p53 activation seen as increased p53 phosphorylation (Ser15). In parallel, increased proteolytic activation of caspase-3 was also observed. Taken together these results suggest that PBR protein expression is directly involved in regulating cell survival and proliferation in human breast cancer cells by influencing signaling mechanisms involved in cell cycle control and apoptosis. PMID- 17126820 TI - Neurotoxic lesions centered on the perifornical hypothalamus abolish the cardiovascular and behavioral responses of conditioned fear to context but not of restraint. AB - The aim of this study was to test the role of the perifornical hypothalamus and adjacent areas in the behavioral and cardiovascular responses to two forms of stress, conditioned fear to context and restraint. Of particular interest was the role of the hypocretin (orexin) containing neurons in these responses. Rats implanted with radio-telemetric probes and fear conditioned to a context received bilateral injections of the neurotoxin hypocretin-2-saporin centered on the perifornical area. One week later, the animals were tested for conditioned fear to context and restraint while recording freezing, 22 kHz ultrasonic vocalizations, activity, mean arterial pressure and heart rate. Histological verification revealed that the lesions were not specific since virtually all the neurons within the injection area were lost. Nevertheless, these lesions, which were centered on the perifornical area, markedly reduced all recorded components of the contextual fear response (by 70%) but had no effect on the response to restraint. The lesions also caused a reduction in body weight and reduced the circadian rhythm of the recorded parameters. The results show (i) that hypocretin 2-saporin was not specific enough to produce lesions restricted to the hypocretin system, (ii) that neurons of the perifornical area are necessary for the expression of both the cardiovascular and behavioral components of conditioned fear to context, and (iii) that the same neurons are not necessary for the cardiovascular response to restraint. Thus, the perifornical hypothalamus is critical for some forms of stress but not others. We propose that it is a necessary relay for emotional responses in which the psychological component is stronger than the sensory component. PMID- 17126822 TI - Role of malonyl-CoA in heart disease and the hypothalamic control of obesity. AB - Obesity is an important contributor to the risk of developing insulin resistance, diabetes, and heart disease. Alterations in tissue levels of malonyl-CoA have the potential to impact on the severity of a number of these disorders. This review will focus on the emerging role of malonyl-CoA as a key "metabolic effector" of both obesity and cardiac fatty acid oxidation. In addition to being a substrate for fatty acid biosynthesis, malonyl-CoA is a potent inhibitor of mitochondrial carnitine palmitoyltransferase (CPT) 1, a key enzyme involved in mitochondrial fatty acid uptake. A decrease in myocardial malonyl-CoA levels and an increase in CPT1 activity contribute to an increase in cardiac fatty acid oxidation. An increase in malonyl-CoA degradation due to increased malonyl-CoA decarboxylase (MCD) activity may be one mechanism responsible for this decrease in malonyl-CoA. Another mechanism involves the inhibition of acetyl-CoA carboxylase (ACC) synthesis of malonyl-CoA, due to AMP-activated protein kinase (AMPK) phosphorylation of ACC. Recent studies have demonstrated a role of malonyl-CoA in the hypothalamus as a regulator of food intake. Increases in hypothalamic malonyl CoA and inhibition of CPT1 are associated with a decrease in food intake in mice and rats, while a decrease in hypothalamic malonyl-CoA increases food intake and weight gain. The exact mechanism(s) responsible for these effects of malonyl-CoA are not clear, but have been proposed to be due to an increase in the levels of long chain acyl CoA, which occurs as a result of malonyl-CoA inhibition of CPT1. Both hypothalamic and cardiac studies have demonstrated that control of malonyl CoA levels has an important impact on obesity and heart disease. Targeting enzymes that control malonyl-CoA levels may be an important therapeutic approach to treating heart disease and obesity. PMID- 17126821 TI - Involvement of chloride channels in IGF-I-induced proliferation of porcine arterial smooth muscle cells. AB - OBJECTIVE: The existence of Cl- channels in vascular smooth muscle cells (VSMCs) has been increasingly investigated, but the biological functions are not yet clear. Insulin-like growth factor (IGF)-I affects proliferation and migration of VSMCs, and dysregulation of this axis may be involved in atherogenesis and intimal hyperplasia. We examined the effects of Cl- channel blockers on IGF-I induced proliferation in porcine VSMCs. The siRNA approach was used to support the role of ClC-2, a member of the volume-regulated Cl- channel family, in cell proliferation of VSMCs. METHODS AND RESULTS: The IGF-I-induced VSMC proliferation was significantly suppressed by the Cl- channel blockers NPPB and IAA94 but not by DIDS. IGF-I-induced cell proliferation parallels a significant increase in the endogenous expression of ClC-2 mRNA and protein. Inhibitors of PI3-kinase, LY294002 and wortmannin, significantly attenuated the IGF-I-upregulated ClC-2 expression and cell proliferation. We observed ClC-2-like Cl- current, and this current was augmented by IGF-I. SiRNA specifically targeted to ClC-2 abolished IGF-I-induced cell proliferation. CONCLUSION: Our data demonstrate that ClC-2 plays a role in IGF-1-induced regulation of VSMC proliferation in cardiovascular diseases. PMID- 17126823 TI - Novel carbonyl and nitrile products from reactive chlorinating species attack of lysosphingolipid. AB - Lysosphingolipids are important lipid signaling molecules that are associated predominantly with high density lipoproteins (HDL) in human plasma. Further, HDL has been shown to be a target for the reactive chlorinating species (RCS) produced by myeloperoxidase (MPO). Accordingly, RCS attack of lysosphingolipids was characterized in these studies. It was shown that RCS attack of sphingosylphosphorylcholine results in the formation of 2-hexadecenal and 1-cyano methano phosphocholine. The structures were identified and confirmed predominantly using mass spectrometric analyses. Further, it was demonstrated that RCS attack of another bioactive lysosphingolipid sphingosine 1-phosphate also results in the formation of 2-hexadecenal from its sphingosine base. Using a synthetically prepared, deuterated 2-hexadecenal internal standard, it was determined that 2-hexadecenal quickly accumulated in HDL treated with MPO/RCS generating system. Thus, the present studies characterize the formation of a novel group of lipid products generated following RCS attack of lysosphingolipids. PMID- 17126824 TI - Effects of selective LDL apheresis on plasma concentrations of ICAM-1, VCAM-1 and P-selectin in diabetic patients with arteriosclerosis obliterans and receiving maintenance hemodialysis. AB - BACKGROUND: Arteriosclerosis obliterans (ASO) is a serious complication in patients with end-stage renal disease (ESRD) caused by diabetic nephropathy. Adsorption of low-density lipoprotein (LDL) has been performed to treat ASO. While efficacy of this treatment has been reported in limb ischemia, the mechanism underlying the benefit remains unclear. We investigated how LDL adsorption affected soluble adhesion molecules; P-selectin, an endothelial and platelet activation marker; inflammatory cytokines such as interleukin (IL) 1beta, IL-6 and tissue necrosis factor (TNF)-alpha; and lipids in serum. METHODS: Selective LDL adsorption by dextran sulfate columns (LDL apheresis) was performed weekly for 10 weeks to treat eight hemodialysis patients with ASO, ESRD, and type 2 diabetes mellitus. Serum was sampled before and immediately after apheresis. RESULTS: LDL apheresis was performed safely. After LDL apheresis lipid concentrations were significantly reduced and clinical findings, such as Fontaine's classification and ankle brachial pressure index values, were improved. Pretreatment concentrations of soluble intercellular and vascular cell adhesion molecules (sICAM-1 and sVCAM-1) and also P-selectin were higher in patients than healthy controls. After apheresis these decreased, especially P selectin. IL-1beta, IL-6, and TNF-alpha concentrations before apheresis were similar to those in controls and were unaffected by treatment. CONCLUSION: Effectiveness of LDL apheresis against ASO may involve decreased endothelial cell and platelet activation. PMID- 17126825 TI - Comparison of access ultrasensitive human growth hormone assay to monoclonal antibody-based immunoradiometric assay. PMID- 17126826 TI - The Oikopleura coenocyst, a unique chordate germ cell permitting rapid, extensive modulation of oocyte production. AB - The ability to adjust reproductive output to environmental conditions is important to the fitness of a species. The semelparous, chordate, Oikopleura dioica, is particularly adept in producing a highly variable number of oocytes in its short life cycle. Here we show that this entails an original reproductive strategy in which the entire female germline is contained in a single multinucleate cell, the "coenocyst". After an initial phase of syncytial nuclear proliferation half of the nuclei entered meiosis whereas the other half became highly polyploid. The inner F-actin network, with associated plasma membranes, formed a highly ramified infrastructure in which each meiotic nucleus was contained in a pseudo-compartmentalized pro-oocyte linked to the common cytoplasm via ring canals. At a set developmental time, a subset of the pro-oocytes was selected for synchronous growth and the common coenocyst cytoplasm was equally partitioned by transfer through the ring canals. Examination of related species indicated that the coenocyst arrangement is a conserved feature of Appendicularian oogenesis allowing efficient numerical adjustment of oocyte production. As Appendicularia are the second most abundant class of zooplankton, with a world-wide distribution, the coenocyst is clearly a common and successful reproductive strategy on a global scale. PMID- 17126827 TI - Striatal serotonin depletion facilitates rat egocentric learning via dopamine modulation. AB - Egocentric spatial learning has been defined as the ability to navigate in an environment using only proprioceptive information, thereby performing a motor response based on one's own movement. This form of learning has been associated with the neural memory system, including the striatum body. Cerebral serotonin depletion induces better performance, both in tasks with strong egocentric components and in egocentric navigation in the Morris' maze. Based on this, we propose that the striatal serotonergic depletion must facilitate egocentric learning. Fifteen female Sprague Dawley rats weighing 250-350 g and maintained under standard conditions were chronically implanted with infusion cannulas for bilateral application of drugs into the striatum. The animals were evaluated for egocentric navigation using the Morris' maze, under different conditions: saline solution infusion, serotonin depletion by infusion of 5,7-Dihydroxytryptamine (25 microg of free base solved in 2.5 microl of ascorbic acid 1% in saline solution), infusion of mixed dopamine D(1) and D(2) receptor antagonists (0.5 microl/min during 5 min of mixed spiperone 20 microM and SCH23390 10 microM), or serotonin depletion and dopamine blockade simultaneously. Striatal serotonin depletion facilitated egocentric learning, which was demonstrated as shorter escape latencies and the display of a defined sequence of movements for reaching the platform. The facilitation was not observed under condition of simultaneous dopamine blockade. Striatal serotonin depletion produced a dopamine-dependent facilitation of egocentric learning. A role for serotonin in the inhibition of striatal-mediated learning strategies is proposed. PMID- 17126828 TI - Additive interaction of intraperitoneal dexmedetomidine and topical nimesulide, celecoxib, and DFU for antinociception. AB - Nimesulide, celecoxib, and DFU (5, 5-dimethyl-3-(3-fluorophenyl)-4-(4 methylsulphonyl)phenyl-2(5H)-furanone) are nonsteroidal anti-inflammatory drugs (NSAIDs) with selective cyclo-oxygenase (COX)-2 blocking properties and have potent analgesic and anti-inflammatory activities in oral and parenteral administrations. Dexmedetomidine, a highly selective alpha(2)-adrenoceptor agonist, is an extremely potent antinociceptive agent. The present study was conducted to evaluate the antinociception induced by nimesulide, celecoxib, and DFU when topically applied on the tail in the absence or presence of intraperitoneal dexmedetomidine. Antinociception was measured in the radiant tail flick test after immersion of the tail of rat into a solution of dimethyl sulfoxide (DMSO) containing nimesulide, celecoxib, or DFU. Antinociceptive effect of all drugs peaked at 60 min and decreased gradually to baseline levels at 240 min. Nimesulide had a potency lower than those of celecoxib, and DFU. The antinociceptive effect of dexmedetomidine was blocked by systemic pretreatment of selective alpha(2)-adrenoceptor antagonist, atipamezole. This suggests that antinociceptive effects of dexmedetomidine involve alpha(2)-adrenoceptors. Combination of topical COX-2 inhibitors with intraperitoneal dexmedetomidine yielded additive analgesic effect. These results demonstrate an additive interaction between topical COX-2 inhibitors with intraperitoneal dexmedetomidine. These observations are significant for physicians to combine selective COX-2 inhibitors and dexmedetomidine in the management of pain. PMID- 17126829 TI - Caffeine induces a profound and persistent tachycardia in response to MDMA ("Ecstasy") administration. AB - Caffeine promotes hyperthermia and lethality when co-administered with the recreational drug 3,4-methylenedioxymethamphetamine (MDMA, "Ecstasy") to rats. In the present study, co-administration of caffeine (10 mg/kg, s.c.) with MDMA (10 mg/kg, s.c.) induced a profound tachycardic response compared to rats treated with either drug alone. However, neither caffeine (30 microM) nor MDMA (1-30 microM), alone or in combination, affected the electrocardiogram of the isolated heart suggesting that central and sympathomimetic actions, rather than direct actions of these drugs on the heart, are responsible for the tachycardia observed in vivo. This is a serious drug interaction, which could have important health consequences for recreational drug users. PMID- 17126830 TI - MHC class II genes in Mexican patients with idiopathic dilated cardiomyopathy. AB - The purpose of the present study was to evaluate the relationship between class II major histocompatibility complex (MHC) genes (HLA-DR and HLA-DQB) and the genetic susceptibility to idiopathic dilated cardiomyopathy (IDC) in Mexican patients. The HLA-DR and DQB alleles were analyzed in 53 patients with IDC and 99 ethnically matched healthy controls using the polymerase chain reaction-sequence specific oligonucleotides (PCR-SSO) technique. IDC patients showed increased frequencies of HLA-DR4 (pC=0.02, OR=1.87), HLA-DQB1*0301 (pC=0.02, OR=1.92) and HLA-DQB1*0302 (pC=0.02, OR=1.87) when compared to healthy controls. On the other hand, IDC patients also showed decreased frequencies of HLA-DR11 allele (pC=0.03, OR=0.26) and HLA-DQB1*0201 (pC=0.04, OR=0.41). These data suggest that variation in class II HLA alleles could be a genetic factor involved in the susceptibility to IDC of the Mexican Mestizo population. PMID- 17126831 TI - Platelets, thrombospondin-1 and human dermal fibroblasts cooperate for stimulation of endothelial cell tubulogenesis through VEGF and PAI-1 regulation. AB - During cutaneous wound repair, platelets, dermal fibroblasts (DF) and endothelial cells all cooperate. We have presently investigated the regulation of endothelial cell tubulogenesis by human platelet thrombospondin-1 (TSP-1), in comparison to transforming growth factor-beta1 (TGF-beta1) and total platelet lysates (PL), in a fibrin matrix cell culture system incorporating DF. TSP-1, TGF-beta1 and PL all stimulated VEGF expression in DF dose dependently at mRNA and protein level. TSP 1- and PL-treated DF supernatants significantly stimulated capillary-like structure formation (tubulogenesis) by dermal microvascular endothelial cells (HMEC-1 and HDMEC), in part via VEGF, as confirmed with neutralizing anti-VEGF antibodies. In contrast, TGF-beta1-treated DF supernatants did not induce tubulogenesis. This apparent discrepancy could be explained by the differential expression regulation in HMEC-1 of fibrinolysis and metalloproteinase mediators by TSP-1 and TGF-beta1. TSP-1 potently reduced the expression of plasminogen activator inhibitor-1 (PAI-1) (mRNA and protein), whereas TGF-beta1 enhanced it. The crucial role of PAI-1 in tubulogenesis was confirmed via the addition of active recombinant PAI-1, which abrogated tubulogenesis. In contrast, neutralizing PAI-1 antibodies enhanced tubulogenesis. Our results suggest that platelet TSP-1 released in a wound stimulates endothelial cell tubulogenesis through an upregulation of DF VEGF expression and a downregulation of endothelial cell PAI-1 expression. PMID- 17126832 TI - Keratin 8 modulation of desmoplakin deposition at desmosomes in hepatocytes. AB - Keratins, the intermediate filament proteins of epithelial cells, connect to desmosomes, the cell-cell adhesion structures at the surface membrane. The building elements of desmosomes include desmoglein and desmocollin, which provide the actual cell adhesive properties, and desmoplakins, which anchor the keratin intermediate filaments to desmosomes. In the work reported here, we address the role of keratin 8 in modulating desmoplakin deposition at surface membrane in mouse hepatocytes. The experimental approach is based on the use of keratin 8- and keratin 18-null mouse hepatocytes as cell models. In wild-type mouse hepatocytes, desmoplakin is aligned with desmoglein and keratin 8 at the surface membrane. In keratin 8-null hepatocytes, the intermediate filament loss leads to alterations in desmoplakin distribution at the surface membrane, but not of desmoglein. Intriguingly, a significant proportion of keratin 18-null hepatocytes express keratin 8 at the surface membrane, associated with a proper desmoplakin alignment with desmoglein at desmosomes. A Triton treatment of the monolayer reveals that most of the desmoplakin present in either wild-type, keratin 8- or keratin 18-null hepatocytes is insoluble. Deletion analysis of keratin 8 further suggests that the recovery of desmoplakin alignment requires the keratin 8 rod domain. In addition, similarly to other works revealing a key role of desmoplakin phosphorylation on its interaction with intermediate filaments, we find that the phosphorylation status of the keratin 8 head domain affects desmoplakin distribution at desmosomes. Together, the data indicate that a proper alignment/deposition of desmoplakin with keratins and desmoglein in hepatocytes requires keratin 8, through a reciprocal phosphoserine-dependent process. PMID- 17126833 TI - Dexamethasone inhibition of trabecular meshwork cell phagocytosis and its modulation by glucocorticoid receptor beta. AB - Glucocorticoid treatment can lead to the development of glaucomatous ocular hypertension and a secondary open-angle glaucoma due to increased aqueous humor outflow resistance that is associated with morphological and biochemical changes in the trabecular meshwork (TM). The cellular responses of glucocorticoids are achieved by binding to the glucocorticoid receptor alpha (GRalpha), a ligand activated transcription factor. An alternatively spliced variant, glucocorticoid receptor beta (GRbeta), has dominant negative activity on GRalpha and has been implicated in a variety of steroid-resistant diseases. We previously showed that GRbeta can block dexamethasone (DEX) responsiveness in TM cells. TM cells are actively phagocytic and function in the removal of debris, pigment and other materials from the aqueous outflow drainage pathway. A decrease in phagocytic activity has been proposed in the pathogenesis of glaucoma and glucocorticoid induced glaucoma. In this study, we investigated the effect of DEX and GRbeta on phagocytosis in normal and glaucomatous TM cells. Human transformed normal NTM-5 and primary normal NTM174-00 cells, which express relatively high amounts of GRbeta, and transformed glaucomatous GTM-3 and primary glaucomatous GTM520-05 cells, which have lower GRbeta expression, were treated with 100 nM DEX or vehicle control for 24h. NTM cells also were transfected with a control or GRbeta expression plasmid to examine the effect of GRbeta on phagocytic activity. The cells were incubated with Alexa 488 conjugated Staphylococcus aureus bioparticles opsonized with rabbit IgG for 1h, followed by fixation and incubation with Alexa 633 conjugated goat anti-rabbit IgG to distinguish ingested from extracellular bioparticles. DAPI nuclear staining was used to quantify cell numbers. Cells and bioparticles were visualized by confocal microscopy. We found that NTM-5 cells ingested more bioparticles than GTM-3 cells. DEX treatment significantly decreased the phagocytosis of bioparticles in NTM-5 and GTM-3 cells, while GTM-3 cells were more responsive to DEX, compared to NTM-5 cells. In primary cell culture, NTM174-00 also engulfed more bioparticles than GTM520-05 cells. DEX treatment significantly decreased the phagocytic activity in GTM520-05, but not in NTM174-00 cells. Transient transfection of pCMX-hGRbeta plasmid increased the expression of GRbeta and consequently maintained the phagocytotic activity of NTM 5 cells in the presence of DEX. Our data demonstrated that the expression level of GRbeta in TM cells can regulate DEX-induced suppression of phagocytotic activity. The lower expression of GRbeta in glaucomatous TM cells may contribute to the altered phagocytic function of TM cells, and may lead to the increased aqueous humor outflow resistance mediated by glucocorticoids. PMID- 17126834 TI - Presence of alpha-globin mRNA and migration of bone marrow cells after sciatic nerve injury suggests their participation in the degeneration/regeneration process. AB - We have previously reported that in the distal stump of ligated sciatic nerves, there is a change in the distribution of myelin basic protein (MBP) and P0 protein immunoreactivities. These results agreed with the studies of myelin isolated from the distal stump of animals submitted to ligation of the sciatic nerve, showing a gradual increase in a 14 kDa band with an electrophoretic mobility similar to that of an MBP isoform, among other changes. This band, which was resolved into two bands of 14 and 15 kDa using a 16% gel, was found to contain a mixture of MBP fragments and peptides with great homology with alpha- and beta-globins. In agreement with these results, we have demonstrated that the mRNA of alpha-globin is present in the proximal and distal stumps of the ligated nerve. It is also detected at very low levels in Schwann cells isolated from normal nerves. These results could be due to the presence of alpha- and/or beta globin arising from immature cells of the erythroid series. Also, they could be present in macrophages, which spontaneously migrate to the injured nerve to promote the degradation of myelin proteins. Cells isolated from normal adult rat bone marrow which were injected intraortically were found to migrate to the injured area. These cells could contribute to the remyelination of the damaged area participating in the removal of myelin debris, through their transdifferentiation into Schwann cells or through their fusion with preexisting Schwann cells in the distal stump of the injured sciatic nerve. PMID- 17126835 TI - Role of microglia in the pathogenesis of osmotic-induced demyelination. AB - Osmotic demyelination is a serious disease caused by rapid correction of hyponatremia. In humans, demyelinative lesions occur preferentially in the central pons, and thus are termed central pontine myelinolysis. Although accumulation of microglia has been reported in such demyelinative lesions, their role in the pathogenesis of osmotic demyelination remains unclear. We examined the expression of cytokines in microglia that accumulated in the demyelinative lesions in a rat model of osmotic demyelination. Hyponatremia was induced in rats by a combination of dDAVP infusion and liquid diet feeding. After 7 days, serum sodium levels were rapidly corrected by hypertonic saline injection. The rats developed severe motor deficits, and marked demyelinative lesions were found in the midbrain and cerebral cortex. In the area of the demyelinative lesions, massive accumulations of microglia were observed that expressed the proinflammatory cytokines TNF-alpha and IFN-gamma as well as iNOS. In contrast, in hyponatremia corrected rats treated with lovastatin, which is known to inhibit microglial infiltration in various animal models of CNS disease, neurological impairments and the degree of demyelination were significantly ameliorated. Lovastatin also reduced the accumulation of microglia and decreased the expression of TNF-alpha in the demyelinative lesions. These results indicate that microglia play a detrimental role in the pathogenesis of osmotic demyelination by producing proinflammatory cytokines, and further suggest that lovastatin may be useful in repressing the demyelination. PMID- 17126836 TI - Effects of frozen storage on the structure of sarcocysts in pig muscle and implications in taxonomic studies. AB - The morphology of the cyst wall of Sarcocystis has unique characteristics that can be used in species identification. To find a suitable way to preserve Sarcocystis cyst samples for species identification, by light microscopy and electron microscopy, we recorded the morphological changes in the cysts of Sarcocystis suihominis and Sarcocystis miescheriana from pig muscle, induced by storage at -20 degrees C. Comparisons were made between fresh cysts and those subjected to frozen storage for periods of 3 days, 20 days and 30 days. RESULTS: cyst wall of the two Sarcocystis species appeared unaffected by storage. There was no obvious change in the length, nor in the width of the protrusions after storage (P>0.05), but the structure of the bradyzoite in the sarcocyst was in many cases disintegrated at -20 degrees C in 20 days for S. miescheriana and 30 days for S. suihominis. To our knowledge this is the first report that Sarcocystis cyst in muscle can be stored at -20 degrees C before and remain suitable for ultrastructural morphological study. Consequently, this paper proposes freezing as a convenient storage method for samples used in taxonomic studies of Sarcocystis. PMID- 17126837 TI - Androgen-sensitivity of somata and dendrites of spinal nucleus of the bulbocavernosus (SNB) motoneurons in male C57BL6J mice. AB - In rats, androgens in adulthood regulate the morphology of motoneurons in the spinal nucleus of the bulbocavernosus (SNB), including the size of their somata and the length of their dendrites. There are conflicting reports about whether androgens exert similar influences on SNB motoneurons in mice. We castrated or sham-operated C57BL6J mice at 90 days of age and, thirty days later, injected cholera toxin conjugated horseradish peroxidase into the bulbocavernosus muscle (to label SNB motoneurons) on one side, and into intrinsic foot muscles contralaterally (to label motoneurons of the retrodorsolateral nucleus (RDLN)). Castrated mice had significantly smaller SNB somas compared to sham-operated mice while there were no differences in soma size of RDLN motoneurons. Dendritic length in C57BL6J mice, estimated in 3-dimensions, also decreased significantly after adult castration. In rats, androgens act directly through androgen receptors (AR) in SNB motoneurons to control soma size and nearly all SNB motoneurons contain AR. Since SNB somata in C57BL6J mice shrank after adult castration, we used immunocytochemistry to characterize AR expression in SNB cells as well as motoneurons in the RDLN and dorsolateral nucleus (DLN). A pattern of labeling matched that seen previously in rats: the highest percentage of AR-immunoreactive motoneurons are in the SNB (98%), the lowest in the RDLN (25%) and an intermediate number in the DLN (78%). This pattern of AR labeling is consistent with the possibility that androgens also act directly on SNB motoneurons in mice to regulate soma size in mice. PMID- 17126838 TI - Testosterone levels and gular pouch coloration in courting magnificent frigatebird (Fregata magnificens): variation with age-class, visited status and blood parasite infection. AB - Male magnificent frigatebirds (Fregata magnificens) possess a seasonally expressed skin ornament, namely the red and inflatable gular pouch, and are, therefore, a convenient model for the study of some theories related to the evolution of possible testosterone-dependent sexual skin coloration. Here we report the findings of a study performed over four consecutive mating seasons in the Mexican national park Isla Isabel. We investigated differences in testosterone level and gular pouch coloration in courting males in relation to the categories: age-class, visited status and blood parasite infection. Gular pouch color saturation increased with age-class. Investigated frigatebirds were infected only with Haemoproteus iwa (Haemosporida, Haemoproteidae), with an overall prevalence infection of 15.5%. Prevalence of the infection increased with birds' age-class. Testosterone levels were significantly higher in infected males, who also had lighter colored gular pouches. In non-infected males, those visited by a female had higher testosterone levels than non-visited males. Gular pouch lightness and redness were negatively correlated but only redness in non visited non-infected males was positively correlated with testosterone levels. Gular pouch saturation in visited and infected males was positively correlated with body mass, which also increased with age-class. Mated males had lower testosterone levels and lighter, less red and saturated gular pouch coloration than courting males. In summary, we found that coloration of the male skin ornament could reflect age-class (saturation), parasite infection (lightness) and mated status (all), together with indications of condition and testosterone dependency of ornament expression. PMID- 17126839 TI - I can C clearly now the rail has gone! AB - Accurate intra-operative imaging is vital in orthopaedic pelvic surgery in order to achieve precise reduction of fractures and correct positioning of osteotomies. Radiological assessment is carried out, in most centres, using standard C-arm fluoroscopy. A limitation of this technique is that it is not always possible to obtain accurate images in all planes due to the presence of radiopaque materials on the operating table. One such example is the 45 degrees oblique view, which is frequently obscured by the presence of a metal rail on the operating table. We report on a practical solution; the development of a unique 360 degrees radiolucent table by placing a sheet of Perspex material between two standard operating tables, which allows for perfect radiological exposure of the pelvis in all planes. PMID- 17126840 TI - Catheter-based adenovirus-mediated anti-monocyte chemoattractant gene therapy attenuates in-stent neointima formation in cynomolgus monkeys. AB - We have previously demonstrated great benefit from anti-monocyte chemoattractant protein-1 (MCP-1) gene therapy by "systemic" transfer of an N-terminal deletion mutant of human MCP-1 (called 7ND) gene into skeletal muscle for treatment of restenosis and atherosclerosis. However, recent evidence suggests that "local" gene transfer may be a clinically relevant approach. We therefore tested the hypothesis that catheter-based adenovirus-mediated anti-MCP-1 gene therapy attenuates stent-associated neointima formation. Bare metal stents were implanted in iliac arteries of cynomolgus monkeys fed a high cholesterol diet. Immediately after the stenting procedure, normal saline or recombinant adenoviral vector containing LacZ or the 7ND gene was administered locally into the stenting site through a Remedy channel-delivery catheter. Compared to saline infusion or LacZ gene transfer, 7ND gene transfer markedly reduced inflammatory changes at an early stage and attenuated neointima formation after 4 weeks. This strategy also reduced the increased production of pro-inflammatory and growth-promoting factors such platelet-derived growth factor. No systemic adverse effects of 7ND gene transfer were detected. There were no significant differences in serum cholesterol levels among the three groups. These data suggest that catheter-based adenovirus-mediated anti-MCP-1 gene therapy may be a clinically relevant and feasible strategy for treatment of in-stent restenosis. PMID- 17126841 TI - Synergistic effect of cytochrome P450 epoxygenase CYP2J2*7 polymorphism with smoking on the onset of premature myocardial infarction. AB - OBJECTIVES: Cytochrome P450 (CYP) 2J2 is expressed in vascular endothelium and metabolizes arachidonic acid to biologically active epoxyeicosatrienoic acids (EETs), which are potent endogenous vasodilators and inhibitors of vascular inflammation. We aimed to elucidate the relationship between the functional CYP2J2*7 polymorphism and smoking for the onset of premature myocardial infarction (MI). PATIENTS/METHODS: We studied 200 patients with acute MI onset under 45 years (84% men) and 200 sex- and age-matched controls. The polymorphism was determined using PCR and direct DNA sequencing analysis. RESULTS: The CYP2J2*7 GT+TT genotype was significantly more prevalent in premature MI patients (32.0% versus 22.0%; p=0.02). Multiple logistic regression analysis showed four independent risk factors: the CYP2J2*7 T allele (OR 1.78, 95% confidence interval [CI] 1.1-6.4; p=0.02), smoking (OR 3.05, 95% CI 1.6-7.3; p<0.01), diabetes mellitus (OR 3.24, 95% CI 1.2-6.6; p<0.01), and hypertension (OR 1.95, 95% CI 1.1 5.7; p<0.01). Among non-smoking patients, the CYP2J2*7 T allele was associated with a 1.3-fold risk. However, smoking T-allele carriers had a significantly 6.7 fold higher risk (p=0.01 for interaction). This variant, but not wild type, significantly reduced promoter activity with nicotine in vitro. EET metabolites were significantly lower among CYP2J2*7 T allele carriers than the GG subjects (p<0.05). Smoking could further lower EET concentrations in T allele carriers than the non-smokers, especially in MI patients (3.3+/-1.0 ng/mL versus 6.8+/-1.3 ng/mL; p=0.001). CONCLUSIONS: The CYP2J2*7 polymorphism and premature MI were synergistically and significantly associated in Taiwanese patients. PMID- 17126842 TI - Electrophoretic determination of albumin in urine using on-line concentration techniques. AB - To improve the sensitivity of the UV-detection for the determination of trace amounts of albumin by capillary zone electrophoresis (CZE), five on-line preconcentration techniques, including field-amplified sample stacking (FASS), head-column field-amplified sample stacking (HC-FASS), stacking with a polymer solution, dynamic pH junction and large volume sample stacking (LVSS) with reversed polarity, were compared. Sensitivity enhancement factor and reproducibility were two factors that were used to assess the suitability of each method. To minimize protein adsorption on the capillary wall, capillaries were covalently modified with anionic polymer, poly(sulfopropylmethacrylate) coating. All used methods have good reproducibility. The maximum sensitivity enhancement factor (about 67-fold in terms of peak heights) was achieved with LVSS technique. The concentration limit of detection (LOD) (S/N=3) for the human serum albumin obtained with the optimized LVSS approach was 15 microg/ml with UV-detection. The method was further evaluated for the analysis of urine samples with gel filtration-based sample-desalting procedure. PMID- 17126843 TI - Optimization of luteolin separation from pigeonpea [Cajanus cajan (L.) Millsp.] leaves by macroporous resins. AB - In the present study, the performance and separation characteristics of eight macroporous resins for the separation of luteolin (LU) from pigeonpea leaves extracts have been evaluated. The adsorption and desorption properties of LU on macroporous resins including AB-8, NKA-9, NKA-2, D3520, D101, H1020, H103 and AL 2 have been compared. AL-2 resin offers the best adsorption and desorption capacity for LU than other resins based on the research results, and its adsorption data at 25 degrees C fit best to the Freundlich isotherm. Dynamic adsorption and desorption experiments have been carried out with the column packed by AL-2 resin to optimize the separation process of LU from pigeonpea leaves extracts. The optimum parameters for adsorption were sample solution LU concentration 65.5 microg/ml, pH 5, processing volume 3 BV, flow rate 1.5BV/h, temperature 25 degrees C; for desorption were elution solvent ethanol-water (50:50, v/v) 2 BV and followed by ethanol-water (60:40, v/v) 2 BV, and flow rate 1BV/h. After treated with AL-2 resin, the LU content in the product was increased 19.8-fold from 0.129% to 2.55%, with a recovery yield of 78.54%. The results showed that AL-2 resin revealed a good ability to separate LU. Therefore, we conclude that results in this study may provide scientific references for the large-scale LU production from pigeonpea or other plants extracts. PMID- 17126844 TI - Use of new generation poly(styrene-divinylbenzene) resins for gas-phase trapping thermal desorption. Application to the retention of seven volatile organic compounds. AB - Two new generation polymeric resins, Bond Elut ENV (styrene-divinylbenzene) from Varian and LiChrolut EN (ethylvinylbenzene-divinylbenzene) from Merck, commonly used in liquid--solid-phase extraction (SPE) were evaluated as sorbents for gas phase sampling followed by thermal desorption and compared to Tenax TA, a reference sorbent in this kind of applications. The three resins were tested against seven volatile organic compounds (VOCs): 1-octene, ethylbenzene, (p-, m-, o-)xylenes, styrene and 1,4-dichlorobenzene. Elution curves for all compounds were determined at temperatures from 120 to 180 degrees C, and from such curves, different parameters, such as retention factor (k), distribution coefficient (K), height equivalent to a theoretical plate (H), asymmetry factor (Fa) and breakthrough volume (VB) were calculated and extrapolated at room (25 degrees C) and desorption (220 degrees C) temperatures in order to estimate breakthrough and elution volumes. In average, retention in LiChrolut EN is 10 and 200 times stronger than in Bond Elut ENV and Tenax TA, respectively, but its chromatographic behavior is rather poor giving quite asymmetric elution profiles (Fa >1.8 at 120 degrees C). Bond Elut ENV exhibited the best chromatographic behavior, with H values two or five times lower than those of LiChrolut EN or Tenax TA. An additional advantage of the new sorbents is that retention decreases with T much faster than it does in Tenax (8 or 20 times for Bond Elut ENV or LiChrolut EN). Modeling has finally shown that beds with 60-80 (for Bond Elut ENV) or 300-400 (for LiChrolut EN) times less of sorbent have the same retention properties than standard Tenax TA tubes and similar (LiChrolut EN) or five to six times smaller (Bond Elut ENV) elution volumes. These predictions have been experimentally confirmed. PMID- 17126845 TI - Rapid column heating method for subcritical water chromatography. AB - A novel resistive heating method is presented for subcritical water chromatography (SWC) that provides higher column heating rates than those conventionally obtained from temperature-programmed gas chromatography (GC) convection ovens. Since the polarity of water reduces dramatically with increasing temperature, SWC employs column heating to achieve gradient elution. As such, the rate at which the mobile phase is heated directly impacts the magnitude of such gradients applied in SWC. Data from the current study demonstrate that the maximum column heating rate attainable in a typical SWC apparatus (i.e. using a GC convection oven) is around 10 degrees C/min, even at instrument oven settings of over three times this value. Conversely, by wrapping the separation column with ceramic insulation and a resistively heated wire, the column heating rates are increased five-fold. As a result, elution times can be greatly decreased in SWC employing gradients. Separations of standard alcohol test mixtures demonstrate that the retention time of the latest eluting component decreases by 35 to 50% using the prototype method. Additionally, solute retention times in this mode deviate by less than 1% RSD over several trials, which compares very well to those obtained using a conventional GC convection oven. Results suggest that the developed method can be a useful alternative heating technique in SWC. PMID- 17126846 TI - Pore structural characterization of monolithic silica columns by inverse size exclusion chromatography. AB - In this work, a parallel pore model (PPM) and a pore network model (PNM) are developed to provide a state-of-art method for the calculation of several characteristic pore structural parameters from inverse size-exclusion chromatography (ISEC) experiments. The proposed PPM and PNM could be applicable to both monoliths and columns packed with porous particles. The PPM and PNM proposed in this work are able to predict the existence of the second inflection point in the experimental exclusion curve that has been observed for monolithic materials by accounting for volume partitioning of the polymer standards in the macropores of the column. The appearance and prominence of the second inflection point in the exclusion curve is determined to depend strongly on the void fraction of the macropores (flow-through pores), (b) the nominal diameter of the macropores, and (c) the radius of gyration of the largest polymer standard employed in the determination of the experimental ISEC exclusion curve. The conditions that dictate the appearance and prominence of the second inflection point in the exclusion curve are presented. The proposed models are applied to experimentally measured ISEC exclusion curves of six silica monoliths having different macropore and mesopore diameters. The PPM and PNM proposed in this work are able to determine the void fractions of the macropores and silica skeleton, the pore connectivity of the mesopores, as well as the pore number distribution (PND) and pore volume distribution (PVD) of the mesopores. The results indicate that the mesoporous structure of all materials studied is well connected as evidenced by the similarities between the PVDs calculated with the PPM and the PNM, and by the high pore connectivity values obtained from the PNM. Due to the fact that the proposed models can predict the existence of the second inflection point in the exclusion curves, the proposed models could be more applicable than other models for ISEC characterization of chromatographic columns with small diameter macropores (interstitial pores) and/or large macropore (interstitial pore) void fractions. It should be noted that the PNM can always be applied without the use of the PPM, since the PPM is an idealization that considers an infinitely connected porous medium and for materials having a low (<6) pore connectivity the PPM would force the PVD to a lower average diameter and larger distribution width as opposed to properly accounting for the network effects present in the real porous medium. PMID- 17126847 TI - Analysis of bacteria degradation products of methyl parathion by liquid chromatography/electrospray time-of-flight mass spectrometry and gas chromatography/mass spectrometry. AB - The biodegradation of the organophosphorus insecticide methyl parathion (MP) in aqueous environment by bacteria isolated from river sediment has been studied. Two species of bacteria which show strong MP degradation ability are identified as Shewanella and Vibrio parahaemolyticus. The biodegradation of MP proceeded rapidly with the formation of a series of intermediate products, which were analyzed using a combination of GC/MS and HPLC/ESI-TOFMS techniques. The major products tentatively identified include a series of reduced products of MP. Results demonstrate that the coupling of TOFMS to HPLC enhances further the capability of LC-MS in the identification of polar organic species in complex environmental samples. Degradation pathways leading to the formation of these products are proposed which involves first the reduction of nitro to amino group in MP, followed by combination with some intrinsic matters of bacteria. The mechanism and products from biodegradation are quite different from those of photocatalytic process for which the main intermediates included methyl paraoxon and 4-nitrophenol. PMID- 17126848 TI - Quantitative determination of the main aliphatic carboxylic acids in wood kraft black liquors by high-performance liquid chromatography-mass spectrometry. AB - The versatile characterization of organic material and especially of the significant aliphatic hydroxy acids in black liquor is of great importance, for example, in monitoring the progress of the kraft pulping process. This paper describes a simple high-performance liquid chromatographic separation method with atmospheric-pressure chemical ionization mass spectrometry (HPLC-APCI-MS) which was developed for the rapid quantitative analysis of these acids, mainly formed as the alkaline degradation products of feedstock carbohydrates. The fraction of carbohydrate degradation products is mainly composed of hydroxy monocarboxylic and volatile acids (formic and acetic acids) along with lesser amounts of various dicarboxylic acids. This method was thoroughly tested and validated to determine the most abundant nonvolatile low-molecular-mass aliphatic mono- and dicarboxylic acids present in softwood (pine and spruce) and hardwood (birch and aspen) kraft black liquors. This straightforward technique provides, compared to the conventional gas chromatographic methods, some important advantages such as simple sample preparation and a faster analysis time, thus enabling almost real time monitoring of these acids. PMID- 17126849 TI - Use of dielectric relaxation for measurements of surface energy variations during adsorption of water on mordenite. AB - This paper tries to assess simply and quantitatively the link between classical adsorption theories and dielectric spectroscopy, in order to demonstrate that dielectric spectroscopy can be used as a tool of determination of surface energy variations due to movements of charge carriers at the surface of solids. A simple theory is developed to analyze hops of cations at the surface of mordenite, which are detected by complex impedance spectroscopy during adsorption of water. An energy of extraction of the cation can be determined from measurements and its dependence on the quantity of water molecules adsorbed is shown and qualitatively and quantitatively explained, using relationships developed in order to interpret adsorption phenomena generally. The agreement with other determinations of the adsorption energies and solid surface energy is correct. PMID- 17126850 TI - Iron dissolution in aqueous AOT solution. AB - The effects of sodium bis(2-ethylhexyl) sulfosuccinate (AOT) on the electrochemical behavior of iron were studied by potentiodynamic and potentiostatic techniques and open-circuit potential measurements. Experiments were made in both neutral and alkaline AOT solutions (pH 7 and pH 12). It was found that AOT-assisted dissolution is initiated on a passivated iron surface and that the oxidation current leads to the formation of a gel-like film on the electrode surface. This dissolution process was investigated as a function of pH, potential, and electrode rotation rate and the corrosion products were characterized by polarizing microscopy, SEM/EDX, and IR spectroscopy. The gel like material is a mixed NaAOT-Fe(AOT)3 lamellar mesophase and a structure for this mesophase is proposed. PMID- 17126851 TI - Metastable tetragonal phase CdWO4 nanoparticles synthesized with a solvothermal method. AB - CdWO4 has only previously been reported in the monoclinic, or wolframite, phase. Here we report the first metastable, tetragonal or scheelite, CdWO4 nanopowder. The tetragonal CdWO4 was synthesized by a propylene glycol solvothermal method. The scheelite phase is stabilized by a combination of high surface area and surface complexation by the propylene glycol. The CdWO4 is stable at 1 bar to 300 degrees C, and converts back to the monoclinic wolframite phase between 300 and 500 degrees C. The nanopowder exhibits cubic morphology and the average particle size of the nanopowder is around 50 nm. PMID- 17126852 TI - Applying grazing incidence X-ray reflectometry (XRR) to characterising nanofilms on mica. AB - Molecularly smooth mica has hitherto not been widely used as a substrate for the X-ray reflectometry (XRR) technique. That is largely due to the difficulty of achieving flatness over a sufficiently large area of mica. Here we show that this difficulty can be overcome by slightly bending the mica substrate over an underlying cylinder; the enhanced rigidity of the bent mica sheet along the axis of the cylinder provides sufficient flatness along this axis for XRR measurements. To test this method, we have employed it to characterise three types of nanofilms on mica in air: (A) Cr-Au thin films; (B) a surface-grown zwitterionic polymer brush; and (C) a Langmuir-Blodgett (LB) phospholipid monolayer, using a table-top X-ray reflectometer. Fitting the obtained reflectivity curves with the standard Parratt algorithm allows us to extract the structural information of the nanofilms (both thickness and apparent roughness). Our simple method points to how XRR may be exploited as a useful characterisation tool for nanofilms on mica. PMID- 17126853 TI - Single variable domain antibody as a versatile building block for the construction of IgG-like bispecific antibodies. AB - Bispecific antibodies (BsAb) have been traditionally utilized to redirect cytotoxic effector cells and agents to kill tumor cells expressing the target antigens. Recently a new concept is emerging to develop BsAb that simultaneously block the functions of two tumor-associated targets, eg., growth factor receptors, for enhanced antitumor efficacies. Broad clinical applications of BsAb have been, and still are, significantly hampered by the difficulty in producing the materials in sufficient quantity and quality by traditional approaches. Here we describe a recombinant approach for the production of an Fc domain-containing, IgG-like tetravalent BsAb, using a single variable domain (sVD) antibody as a versatile building block. In this method, a sVD of a defined specificity is genetically fused to either the N-terminus of the light chain or the C-terminus of the heavy chain of a functional IgG antibody of a different specificity. A model BsAb was constructed using a sVD to mouse platelet derived growth factor receptor alpha and a conventional IgG antibody to mouse platelet derived growth factor receptor beta. The BsAb were expressed in mammalian cells and purified to homogeneity by a one-step Protein A affinity chromatography. Further, the BsAb retained the antigen binding specificity and the receptor neutralizing activity of both of its parent antibodies. Importantly, the BsAb inhibited the activation of both its target receptors in tumor cells stimulated by both platelet derived growth factor AA and BB, whereas the parent monospecific antibody only inhibited the activation of a single receptor stimulated by its cognate ligand. This format of BsAb should be readily applicable to the production of other BsAb recognizing any pairs of antigens. PMID- 17126854 TI - A fast and efficient procedure to produce scFvs specific for large macromolecular complexes. AB - We have expanded the application of antibody phage display to a new type of antigen: ribonucleoprotein (RNP) complexes. We describe a simple and efficient method for screening antibodies specific for large intact RNPs and individual components. We also describe a fast and easy method to overcome the abundance of amber stop codons in the positive phage clones. The resulting antibodies have been used in ELISA and Western blot analysis. PMID- 17126855 TI - Stage-specific distribution of oxidative radicals and antioxidant enzymes in the midgut of Leptinotarsa decemlineata. AB - The titers of reactive oxygen species (ROS) represented by superoxide anion and general peroxides, and the activities of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), are regulated in the midgut of the Colorado potato beetle (CPB) relative to the gut compartment, developmental stage, and food intake. ROS concentration is low in the potato leaves but it is very high in their digest in insect's anterior midgut. It is proposed that intensive ROS production in this gut region is linked to the processing of allelochemicals. SOD and CAT activities, low oxygen tension, and unidentified redox systems that maintain a slightly reducing milieu in the midgut lumen (pe+pH=6.95 declining to 5.36), obviously contribute to the decrease of ROS concentration along the gut length to a minimum in the wall of posterior midgut region. SOD and CAT activities are higher in the potato leaves than in the midgut tissues but the role of plant enzymes in ROS elimination within the gut lumen remains to be shown. A lower level of ROS and a higher antioxidant potential in the adult than in the larval midgut indicate stage specificity in the management of oxidative stress. The antioxidant defense is high in the diapausing adults that contain no detectable superoxide and about ten times less peroxides than the reproducing adults. PMID- 17126856 TI - Stereoselective reduction of 4-benzoylpyridine in the heart of vertebrates. AB - The stereoselectivity in the reduction of 4-benzoylpyridine (4-BP) was examined in the cytosolic fractions from the heart of 9 vertebrates (pig, rabbit, guinea pig, rat, mouse, chicken, soft-shelled turtle, frog and flounder). 4-BP was stereoselectively reduced to S(-)-alpha-phenyl-4-pyridylmethanol [S(-)-PPOL] in the cytosolic fractions from the heart of pig, rabbit and guinea pig. However, of mammalian heart cytsol tested, only rat heart cytosol had little ability to reduce stereoselectively 4-BP. In an attempt to elucidate this reason, amino acid sequence of rat heart carbonyl reductase (RatHCR) was deduced from the cloned cDNA and compared with that of pig heart carbonyl reductase (PigHCR), which shows a high stereoselectivity in the reduction of 4-BP to S(-)-PPOL. RatHCR showed a high identity with PigHCR in amino acid sequence. Furthermore, recombinant RatHCR was confirmed to reduce stereoselectively 4-BP to S(-)-PPOL with a high optical purity comparable to recombinant PigHCR. It is possible that in the cytosolic fraction from the heart of rat, constitutive reductase other than RatHCR counteracts the stereoselective reduction of 4-BP to S(-)-PPOL, by catalyzing the reduction of 4-BP to the R(+)-enantiomer. PMID- 17126857 TI - Effect of thiazolidinediones on bile acid transport in rat liver. AB - The thiazolidinedione derivatives, troglitazone, rosiglitazone, and pioglitazone, are novel insulin-sensitizing drugs that are useful in the treatment of type 2 diabetes. However, hepatotoxicity associated with troglitazone led to its withdrawal from the market in March 2000. In view of case reports of hepatotoxicity from rosiglitazone and pioglitazone, it is unclear whether thiazolidinediones as a class are associated with hepatotoxicity. Although the mechanism of troglitazone-associated hepatotoxicity has not been elucidated, troglitazone and its major metabolite, troglitazone sulfate, competitively inhibit adenosine triphosphate (ATP)-dependent taurocholate transport in isolated rat canalicular liver plasma membrane vesicles mediated by the canalicular bile salt export pump (Bsep). These results suggest that cholestasis may be a factor in troglitazone-associated hepatotoxicity. To determine whether this effect is 1) limited to canalicular bile acid transport and 2) is specific to troglitazone, the effect of troglitazone, rosiglitazone, and ciglitazone on bile acid transport was examined in rat basolateral (blLPM) and canalicular (cLPM) liver plasma membrane vesicles. In cLPM vesicles, troglitazone, rosiglitazone, and ciglitazone (100 microM) all significantly inhibited ATP-dependent taurocholate transport. In blLPM vesicles, these three thiazolidinediones also significantly inhibited Na(+) dependent taurocholate transport. Inhibition of bile acid transport was concentration dependent and competitive in both cLPM and blLPM vesicles. In conclusion, these findings are consistent with a class effect by thiazolidinediones on hepatic bile acid transport. If hepatotoxicity is associated with this effect, then hepatotoxicity is not limited to troglitazone. Alternatively, if hepatotoxicity is limited to troglitazone, other mechanisms are responsible for its reported hepatotoxicity. PMID- 17126858 TI - The 1974 spill of the Bouchard 65 oil barge: petroleum hydrocarbons persist in Winsor Cove salt marsh sediments. AB - Petroleum hydrocarbons persist in salt marsh sediments in Winsor Cove (Buzzards Bay, Massachusetts) impacted from the 1974 spill of No. 2 fuel oil by the barge Bouchard 65. Intertidal sediment cores were collected from 2001 to 2005 and analyzed for total petroleum hydrocarbons (TPHs). TPHs content was greatest (as high as 8.7 mg g(-1) dry weight) in the surface sediments and decreased with distance landward. Select samples were analyzed for polycyclic aromatic hydrocarbons (PAHs) with values as high as 16.7 microg g(-1) for total naphthalenes and phenanthrenes/anthracenes. These remaining PAHs are mainly C(4) naphthalenes and C(1)-, C(2)-, and C(3)-phenanthrenes/anthracenes revealing preferential loss of almost all of the naphthalenes and the parent compound phenanthrene. Inspection of the data indicates that biodegradation, water-washing and evaporation were major removal processes for many of the petroleum hydrocarbons in the marsh sediments. In addition, historical data and photographs combined with their recent counterparts indicate that erosion has physically removed these contaminants from this site. PMID- 17126859 TI - The effects of the mGluR5 antagonist MPEP and the mGluR2/3 antagonist LY341495 on rats' performance in the 5-choice serial reaction time task. AB - Schizophrenia is characterized by attentional deficits possibly associated with glutamate dysfunction. The role of postsynaptic metabotropic glutamate 5 receptors (mGluR5) or presynaptic inhibitory mGluR2/3 on attention is currently unknown. We investigated the effects of the mGluR5 antagonist MPEP (2-methyl 6[phenylethynyl]-pyridine) and the mGluR2/3 antagonist LY341495 on attention in the 5-choice serial reaction time task (5CSRTT), as well as on food intake to evaluate their effects on food motivation. The effects of pre-feeding and the muscle relaxant curare were examined to characterize the effects of alterations in the motivation or ability to perform the task, respectively. MPEP had no effect on accuracy but overall decreased performance in the 5CSRTT, including decreased speed of responding and decreased premature responses. LY341495 had no significant effect on rats' performance in the 5CSRTT. LY341495 decreased food intake in the home cage to a greater extent than MPEP. Curare decreased the speed of correct responding, reflecting motor impairment. Free feeding decreased overall performance, number of trials completed and number of head entries into the feeder, reflecting decreased motivation to perform the task. Thus, blockade of mGluR5, but not mGluR2/3, decreased overall responding without affecting accuracy in the 5CSRTT. PMID- 17126860 TI - Pharmacological and genetic manipulation of kappa opioid receptors: effects on cocaine- and pentylenetetrazol-induced convulsions and seizure kindling. AB - The present study used pharmacological and gene ablation techniques to examine the involvement of kappa opioid receptors (KOPr) in modulating the convulsant effects of two mechanistically different drugs: cocaine and pentylenetetrazol (PTZ; GABA-A receptor antagonist) in mice. Systemic administration of the selective KOPr-1 agonist, U69593 (0.16-0.6mg/kg; s.c.), failed to modify cocaine evoked convulsions or cocaine kindling. Similarly, no alteration in responsiveness to cocaine was observed in wild-type mice that received the selective KOPr-1 antagonist, nor-binaltorphimine (nor-BNI; 5mg/kg) or in mice lacking the gene encoding KOPr-1. In contrast to cocaine, U69593 attenuated the seizures induced by acute or repeated PTZ administration. Nor-BNI decreased the threshold for PTZ-evoked seizures and increased seizure incidence during the initial induction of kindling relative to controls. Decreased thresholds for PTZ induced seizures were also observed in KOPr-1 knock out mice. Together, these data demonstrate an involvement of endogenous KOPr systems in modulating vulnerability to the convulsant effects of PTZ but not cocaine. Furthermore, they demonstrate that KOPr-1 activation protects against acute and kindled seizures induced by this convulsant. Finally, the results of our study suggest that KOPr-1 antagonists will not have therapeutic utility against cocaine-induced seizures, while they may prove beneficial in attenuating several actions of cocaine that have been linked to its abuse. PMID- 17126861 TI - Disgust and Huntington's disease. AB - The disproportionate impairment for the recognition of facial expressions of disgust in patients with Huntington's disease (HD) forms a double dissociation with the impaired recognition of fear that has been reported in amygdala patients. The dissociation has generated discussion regarding the potential existence of neural substrates dedicated to the recognition of facial signals of specific emotions. The aim of this study was to establish whether the impairment for disgust in HD was restricted solely to the domain of facial perception, or whether HD patients also demonstrate impairment in other kinds of disgust. Fourteen HD patients and fourteen age and education matched healthy controls participated in seven disparate emotion processing tasks. (1) A measure of knowledge for the situational determinants of distinct emotions; (2) recognition of emotion expressed in nonverbal vocalisations; (3) recognition of the emotional content of explicit lexical stimuli; (4) recognition of emotional content in pictures of emotion scenes; (5) a disgust experience questionnaire; (6) a measure of olfactory hedonic responsiveness; (7) a measure of gustatory perception. While verbal aspects of disgust processing were preserved, parallel impairments were revealed for olfactory disgust, vocal disgust expressions, the classification of disgusting pictures, and declarative knowledge of disgust elicitors. The finding of impaired perception of disgust signalled through different input domains suggests that the inability to recognise the facial expression in this population reflects a fundamental problem with disgust processing. PMID- 17126862 TI - Medial septum lesions disrupt exploratory trip organization: evidence for septohippocampal involvement in dead reckoning. AB - Rats organize their open field behavior into a series of exploratory trips focused around a central location or home base. In addition, differences in movement kinematics have been used to fractionate the exploratory trip into tour (i.e., sequences of linear movement or progressions punctuated by stops) and homeward (i.e., single progression direct to the home base) segments. The observation of these characteristics independent of environmental familiarity and visual cue availability has suggested a role for self-movement information or dead reckoning in organizing exploratory behavior. Although previous work has implicated a role for the septohippocampal system in dead reckoning based navigation, as of yet, no studies have investigated the contribution of the medial septum to dead reckoning. First, the present study examined the organization of exploratory behavior under dark and light conditions in control rats and rats receiving either electrolytic or sham medial septum lesions. Medial septum lesions produced a significant increase in homeward segment path circuity and variability of temporal pacing of linear speeds. Second, as an independent assessment of the effectiveness of the medial septum lesions, rats were trained to locate a hidden platform in the standard water maze procedure. Consistent with previous research, medial septum lesions attenuated learning the location of the hidden platform. These results demonstrate a role for the medial septum in organizing exploratory behavior and provide further support for the role of the septohippocampal system in dead reckoning based navigation. PMID- 17126863 TI - Peramine and other fungal alkaloids are exuded in the guttation fluid of endophyte-infected grasses. AB - Many grasses live in association with asymptomatic fungi (Neotyphodium spp. endophytes), which grow in the intercellular spaces of the grass. These endophytes produce a range of alkaloids that protect the grass against grazing by mammals and insects. One of these alkaloids is an unusual pyrrolopyrazine, peramine. Peramine appears to be continuously produced by the endophyte, but does not progressively accumulate. No mechanism for the removal of peramine by its further metabolism or any other process has been reported. Our aim was to detect peramine or peramine metabolites in plant fluids to determine if peramine is mobilized, metabolized or excreted by the plant. We also wanted to determine if other fungal metabolites are mobilized by the plant, as has been proposed for the loline alkaloids. We developed a highly sensitive method for the analysis of peramine, using a linear ion trap mass spectrometer. We studied the fragmentation pathway of peramine using ESI MSn and ESI FTICRMS. Based on these results we developed a single reaction monitoring method using the fragmentation of the guanidinium moiety. Cut leaf fluid and guttation fluid of different grass endophyte associations (Lolium perenne with Neotyphodium lolii, Festuca arundinacea with Neotyphodium coenophialum, and Elymus sp. with Epichloe sp.) were analysed. Peramine was detected in the cut leaf fluid of all grass-endophyte associations, but not in the guttation fluid of all associations. In some associations we also detected lolines and ergot peptide alkaloids. This is the first report showing the mobilization of fungal alkaloids into plant fluids by the host plant in grass-endophyte associations. PMID- 17126864 TI - Elemanolide sesquiterpenes and eudesmane sesquiterpene glycosides from Centaurea hierapolitana. AB - Two elemanolide sesquiterpenes and two eudesmane-type sesquiterpene glycosides named hierapolitanins A-D, were isolated, together with five known compounds, two flavones; hispidulin and jaceosidin, a flavon-C-glycoside, shaftoside, a flavonol glycoside, kaempferol-3-O-rutinoside and a neolignan, dehydrodiconiferyl alcohol from the aerial parts of Centaurea hierapolitana Boiss. (Asteraceae). Structure elucidations were based on spectroscopic evidence. PMID- 17126865 TI - Antiproliferative activity is predominantly associated with ellagitannins in raspberry extracts. AB - Raspberry extracts enriched in polyphenols, but devoid of organic acids, sugars and vitamin C, were prepared by sorption to C18 solid phase extraction matrices and tested for their ability to inhibit the proliferation of human cervical cancer (HeLa) cells in vitro. The raspberry extract reduced proliferation in a dose-dependent manner whether this was judged by cell number or measurements of cell viability. However, measurements based on cell viability were more accurate and gave an EC(50) value of 17.5 microg/ml gallic acid equivalents (GAE) at day 4 of culture. Raspberry extracts were fractionated by sorption to Sephadex LH-20 into an unbound fraction, which was obviously enriched in anthocyanins, and a bound fraction. The unbound anthocyanin-enriched fraction was much less effective in reducing proliferation then the original extract and gave an EC(50) value estimated at 67 microg/ml. The LH-20 bound fraction was more effective than the original raspberry extract (EC(50)=13 microg/ml) suggesting that the main anti proliferative agents were retained in the bound fraction. Analysis of the original extract, the unbound and the LH20 bound fractions by LC-MS confirmed that the unbound fraction was enriched in anthocyanins and the bound fraction primarily contained ellagitannins. The ellagitannin-rich bound fraction had the highest antioxidant capacity as measured by the ferric reducing antioxidant potential (FRAP) assay. The mechanism by which the ellagitannins inhibit proliferation of cancer cells is discussed. PMID- 17126866 TI - Synergism and redundancy in a plant volatile blend attracting grapevine moth females. AB - A flight tunnel study was done to decipher the behavioral effect of grape odor in grapevine moth Lobesia botrana. A blend of 10 volatile compounds, which all elicit a strong antennal response, attracts mated grapevine moth females from a distance, by upwind orientation flight. These 10 grape volatiles are in part behaviorally redundant, since attraction to a 3-component blend of beta caryophyllene, (E)-beta-farnesene and (E)-4,8-dimethyl-1,3,7-nonatriene was not significantly different from the 10-component blend. Blending these three compounds had a strong synergistic effect on female attraction, and omission of any one compound from this 3-component blend almost abolished attraction. It was nonetheless possible to substitute the three compounds with the other grape volatiles which are perceived by the female antenna, to partly restore attraction. Several blends, of varying composition, elicited significant attraction. The observed behavioral plasticity in response to grape volatile blends probably reflects the variation of the natural plant signal, since females oviposit on different grape varieties, in different phenological stages. PMID- 17126867 TI - Epidemiology of invasive pneumococcal disease in Kumasi, Ghana. AB - There are few data on the epidemiology of invasive pneumococcal disease in Africa. We undertook a prospective study of these infections in Kumasi, Ghana, collecting clinical data on all patients with laboratory-confirmed pneumococcal meningitis, pneumonia or systemic sepsis associated with bacteraemia. A total of 140 cases were identified in the period from January 2002 to April 2005. The disease was most prevalent among patients <5 years of age and immediately following the peak of the harmattan wind. The majority of patients were treated with a combination of antibiotics, in part reflecting concerns regarding antibiotic resistance. Mortality was high (47%), with no evidence of an improved prognosis compared with earlier studies in the region. Although most isolates of pneumococci were resistant to tetracyclines and co-trimoxazole, there was no high level resistance to penicillin and only 12% of isolates showed intermediate level resistance. Serotype 1 was the most common serotype (36%), whilst intermediate level penicillin resistance was associated with serotype 14. Theoretical coverage by existing 7-, 9-, 11- and 23-valent vaccines was 26%, 63%, 64% and 76%, respectively. Vaccination may improve control of pneumococcal disease in Ghana, although modified vaccine formulations are required for local use. PMID- 17126868 TI - Impact of climate change on health: what is required of climate modellers? AB - The potential impacts of climate change on human health are significant, ranging from direct effects such as heat stress and flooding, to indirect influences including changes in disease transmission and malnutrition in response to increased competition for crop and water resources. Development agencies and policy makers tasked with implementing adaptive strategies recognize the need to plan for these impacts. However at present there is little guidance on how to prioritize their funding to best improve the resilience of vulnerable communities. Here we address this issue by arguing that closer collaboration between the climate modelling and health communities is required to provide the focused information necessary to best inform policy makers. The immediate requirement is to create multidisciplinary research teams bringing together skills in both climate and health modelling. This will enable considerable information exchange, and closer collaboration will highlight current uncertainties and hopefully routes to their reduction. We recognize that climate is only one aspect influencing the highly complex behaviour of health and disease issues. However we are optimistic that climate-health model simulations, including uncertainty bounds, will provide much needed estimates of the likely impacts of climate change on human health. PMID- 17126869 TI - A comparative immunogenicity study in rabbits of disulfide-stabilized, proteolytically cleaved, soluble trimeric human immunodeficiency virus type 1 gp140, trimeric cleavage-defective gp140 and monomeric gp120. AB - The human immunodeficiency virus type 1 (HIV-1) surface envelope glycoprotein (Env) complex, a homotrimer containing gp120 surface glycoprotein and gp41 transmembrane glycoprotein subunits, mediates the binding and fusion of the virus with susceptible target cells. The Env complex is the target for neutralizing antibodies (NAbs) and is the basis for vaccines intended to induce NAbs. Early generation vaccines based on monomeric gp120 subunits did not confer protection from infection; one alternative approach is therefore to make and evaluate soluble forms of the trimeric Env complex. We have directly compared the immunogenicity in rabbits of two forms of soluble trimeric Env and monomeric gp120 based on the sequence of HIV-1(JR-FL). Both protein-only and DNA-prime, protein-boost immunization formats were evaluated, DNA-priming having little or no influence on the outcome. One form of trimeric Env was made by disrupting the gp120-gp41 cleavage site by mutagenesis (gp140(UNC)), the other contains an intramolecular disulfide bond to stabilize the cleaved gp120 and gp41 moieties (SOSIP.R6 gp140). Among the three immunogens, SOSIP.R6 gp140 most frequently elicited neutralizing antibodies against the homologous, neutralization-resistant strain, HIV-1(JR-FL). All three proteins induced NAbs against more sensitive strains, but the breadth of activity against heterologous primary isolates was limited. When antibodies able to neutralize HIV-1(JR-FL) were detected, antigen depletion studies showed they were not directed at the V3 region but were targeted at other, undefined gp120 and also non-gp120 epitopes. PMID- 17126870 TI - Recruitment of activated IRF-3 and CBP/p300 to herpes simplex virus ICP0 nuclear foci: Potential role in blocking IFN-beta induction. AB - The host innate response to viral infection includes the production of interferons, which is dependent on the coordinated activity of multiple transcription factors. Herpes simplex virus 1 (HSV-1) has been shown to block efficient interferon expression by multiple mechanisms. We and others have demonstrated that HSV-1 can inhibit the transcription of genes promoted by interferon regulatory factor-3 (IRF-3), including interferon beta (IFN-beta), and that the immediate-early ICP0 protein is sufficient for this function. However, the exact mechanism by which ICP0 blocks IRF-3 activity has yet to be determined. Unlike some other viral proteins that inhibit IRF-3 activity, ICP0 does not appear to affect phosphorylation and dimerization of IRF-3. Here, we show that a portion of activated IRF-3 co-localizes with nuclear foci containing ICP0 at early times after virus infection. Co-localization to ICP0-containing foci is also seen with the IRF-3-binding partners and transcriptional co-activators, CBP and p300. In addition, using immunoprecipitation of infected cell lysates, we can immunoprecipitate a complex containing ICP0, IRF-3, and CBP. Thus we hypothesize that ICP0 recruits activated IRF-3 and CBP/p300 to nuclear structures, away from the host chromatin. This leads to the inactivation and accelerated degradation of IRF-3, resulting in reduced transcription of IFN-beta and an inhibition of the host response. Therefore, ICP0 provides an example of how viruses can block IFN beta induction by sequestration of important transcription factors essential for the host response. PMID- 17126871 TI - Stoichiometry of the antiviral protein APOBEC3G in HIV-1 virions. AB - A host cytidine deaminase, APOBEC3G (A3G), inhibits replication of human immunodeficiency virus type 1 (HIV-1) by incorporating into virions in the absence of the virally encoded Vif protein (Deltavif virions), at least in part by causing G-to-A hypermutation. To gain insight into the antiretroviral function of A3G, we determined the quantities of A3G molecules that are incorporated in Deltavif virions. We combined three experimental approaches-reversed-phase high pressure liquid chromatography (HPLC), scintillation proximity assay (SPA), and quantitative immunoblotting-to determine the molar ratio of A3G to HIV-1 capsid protein in Deltavif virions. Our studies revealed that the amount of the A3G incorporated into Deltavif virions was proportional to the level of its expression in the viral producing cells, and the ratio of the A3G to Gag in the Deltavif virions produced from activated human peripheral blood mononuclear cells (PBMC) was approximately 1:439. Based on previous estimates of the stoichiometry of HIV-1 Gag in virions (1400-5000), we conclude that approximately 7 (+/-4) molecules of A3G are incorporated into Deltavif virions produced from human PBMCs. These results indicate that virion incorporation of only a few molecules of A3G is sufficient to inhibit HIV-1 replication. PMID- 17126872 TI - Interocular interactions during acuity measurement in children and adults, and in adults with amblyopia. AB - The binocular interactions that occur during dichoptic and binocular viewing were investigated using a letter acuity task in normally sighted children (age range 6 14 years) and adults, and in adults with anisometropic amblyopia. Our aims were to investigate the nature of binocular interactions that occur in each group, and the extent to which the characteristics of binocular interactions differ across the groups. The non-tested eye was occluded during monocular (baseline) viewing, and was allowed to view a uniform stimulus with fusion lock in dichoptic viewing. In adults and children with normal vision, acuity under dichoptic viewing was unchanged relative to monocular baseline in the dominant eyes, while acuity of the non-dominant eye improved under dichoptic viewing relative to baseline. The magnitude of dichoptic change in the non-dominant eyes was similar in the two normally sighted groups, but the dichoptic advantage was found to decrease with increasing age within the children tested. Binocular acuity was better than monocular acuity in normal subjects, and a decrease in binocular summation with age was noted within the age range of the children tested. In contrast, the amblyopic observers showed no change in acuity with viewing conditions. The results demonstrate development of interocular interactions during childhood, and wide inter-individual variation in pattern of interocular interactions among anisometropic amblyopic adults. PMID- 17126873 TI - Simultaneous nitrification and p-cresol oxidation in a nitrifying sequencing batch reactor. AB - The tolerance, kinetic behavior and oxidizing ability of a nitrifying sludge exposed to different initial concentrations of p-cresol (25-150mg/l) were evaluated in a sequencing batch reactor (SBR) fed with 200mg NH(4)(+)-N/ld. The nitrifying SBR operated up to 300mg/ld of p-cresol, achieving simultaneously the complete ammonium oxidation to nitrate and the total consumption of p-cresol and its transitory intermediates from the culture. p-Cresol induced a significant decrease in the values for specific rates of ammonium consumption, showing that the ammonium oxidation pathway was mainly inhibited. After 7 months of operation in SBR, the specific rates of NH(4)(+)-N oxidation, NO(3)(-)-N formation, and total organic carbon consumption were 0.6g NH(4)(+)-N/g microbial protein-Nh, 0.3g NO(3)(-)-N/g microbial protein-Nh, and 0.24g total organic carbon/g microbial protein h, respectively. The microbial growth rate was always low (maximum value of 12.2+/-0.4mg protein-N/ld) and settleability of the sludge was good with sludge volume index values lower than 21ml/g. The oxidation of p-cresol and its intermediates was carried out faster throughout the cycles and nitrification inhibition decreased with the number of cycles. PMID- 17126874 TI - Fuel-grade ethanol transport and impacts to groundwater in a pilot-scale aquifer tank. AB - Fuel-grade ethanol (76L of E95, 95%v/v ethanol, 5%v/v hydrocarbon mixture as a denaturant) was released at the water table in an 8150-L continuous-flow tank packed with fine-grain masonry sand. Ethanol, which is buoyant and hygroscopic, quickly migrated upwards and spread laterally in the capillary zone. Horizontal migration of ethanol occurred through a shallow thin layer with minimal vertical dispersion, and was one order of magnitude slower than the preceding bromide tracer. Dyes, one hydrophobic (Sudan-IV) and one hydrophilic (Fluorescein) provided evidence that the fuel hydrocarbons phase separated from the E95 mixture as ethanol was diluted by pore water and its cosolvent effect was diminished. Most of the added ethanol (98%) was recovered in the effluent wells that captured the flow through the high water content regions above the water table. Complementary bench-scale 2-D visualization experiments with E95 confirmed hydrocarbon phase separation, residual NAPL formation and migration within the capillary fringe. These results corroborate previous bench-scale studies showing that ethanol has high affinity for vadose-zone pore water and can migrate through the capillary zone. The pilot-scale tank experiment provides the first hydrocarbon and ethanol concentration measurements (and thus, quantification of impacts to groundwater quality) from a subsurface spill of E95 in a well characterized system with a well-defined source. It also provides the first quantitative near-field-scale evidence that capillarity can significantly retard the vertical dispersion and horizontal advection of ethanol. Such effects could be important determinants of the extent of ethanol migration and longevity as well as groundwater impacts. PMID- 17126875 TI - Longitudinal mixing in meandering channels: new experimental data set and verification of a predictive technique. AB - Evaluation of longitudinal mixing processes in open channel flows is important in environmental management, requiring the quantification of mixing coefficients. Estimates of these coefficients sufficiently accurate for environmental impact assessments cannot be achieved using current theoretical or semi-empirical methods for natural channels. This inaccuracy is caused by a limited understanding and quantification of the interaction of the dominant mechanisms resulting from natural channel features, such as plan form curvature and changes in cross-sectional shape. Experimental results are presented here from studies conducted in three self-formed channels, developed by known discharges. Longitudinal mixing was investigated at various flow rates within each of the channels by monitoring the development of tracer plumes during transit through the channels. Using an optimisation procedure, coefficients required for solution of the one-dimensional advection dispersion equation (1D-ADE) were found in the range 0.02-0.2m(2)/s. The coefficients were found to vary as functions of longitudinal meander location, channel form and discharge. Predictions of these longitudinal mixing coefficients were made using a mathematical technique requiring only channel form properties and flow rate as inputs. Predicted values were typically within 20% of the measured values, although deviation of up to 50% was found for the lowest discharge in each channel. This large error is likely to have been caused by increased dead zone effects associated with channel bathymetry at low discharges that are not captured by the method. The method was shown to be capable of capturing the variation in the longitudinal mixing coefficient with longitudinal meander location, with channel form and with discharge. PMID- 17126876 TI - Benthic cyanobacteria (Oscillatoriaceae) that produce microcystin-LR, isolated from four reservoirs in southern California. AB - Cyanobacteria that produce the toxin microcystin have been isolated from many parts of the world. Most of these organisms are planktonic; however, we report on several microcystin-producing benthic filamentous cyanobacterial isolates from four drinking-water reservoirs in southern California (USA): Lake Mathews, Lake Skinner, Diamond Valley Lake (DVL), and Lake Perris. Some samples of benthic material from these reservoirs tested positive for microcystin by an ELISA tube assay, and all the positive samples had in common a green filamentous cyanobacterium 10-15microm in diameter. Seventeen unialgal strains of the organism were isolated and tested positive by ELISA, and 11 cultures of these strains were found to contain high concentrations of microcystin-LR (90 432microgL(-1)). The cultures were analyzed by protein phosphatase inhibition assay (PPIA) and HPLC with photodiode array detector (PDA) or liquid chromatography/mass spectrometry (LC/MS). Microcystin per unit carbon was determined for six cultures and ranged from 1.15 to 4.15microgmg(-1) C. Phylogenetic analysis of four cultures from Lake Skinner and DVL using cyanobacterial-specific PCR and sequencing of the partial 16S rRNA gene suggested the highest similarity to an unidentified cyanobacterium in the oscillatoriales, and to a Phormidium sp. Morphologically, some of the isolates were similar to Oscillatoria, and others resembled Lyngbya. The significance of these organisms lies in the relative scarcity of known toxin producers among freshwater benthic cyanobacteria, and also as a source of cell-bound microcystin in these reservoirs. PMID- 17126877 TI - Phosphate and potassium recovery from source separated urine through struvite precipitation. AB - Phosphate can be recovered as struvite or apatite in fluidised bed reactors. Urine has a much higher phosphate concentration than sludge reject water, allowing simpler (and less expensive) process for precipitation of phosphates. A stirred tank reactor with a special compartment for liquid solid separation was used to precipitate struvite from urine. Magnesium ammonium phosphate as well as potassium magnesium phosphate are two forms of struvite that were successfully precipitated. Liquid/solid separation was very effective, but the compaction of struvite was rather poor in the case of potassium struvite. Crystals did not form clusters and maintained the typical orthorhombic structure. Ammonium struvite had slightly lower effluent phosphate concentrations, but an average of 95% of influent phosphate was removed regardless of ammonium or potassium struvite precipitation. Fluid mechanics is believed to be important and should inform further work. PMID- 17126878 TI - Effects of Cd2+ on K+, Ca2+ and N uptake in two halophytes Sesuvium portulacastrum and Mesembryanthemum crystallinum: consequences on growth. AB - One of the limits of Cd2+-phytoextraction is the high toxicity of this metal to plants. Growth restriction, chlorosis and necrosis are usually accompanied with a large disturbance of the uptake of essential elements. This work aims to study the effects of cadmium (Cd2+) on potassium (K+), calcium (Ca2+) and nitrogen (N) acquisition, and their consequences on growth in two halophytes species: Sesuvium portulacastrum and Mesembryanthemum crystallinum. Seedlings were grown for 30 days in split-root conditions. One half of the root system was immersed in complete nutrient solution (Basal medium (B)) supplemented with 100 microM Cd2+, and the other half was immersed in a Cd2+-free medium, containing all nutrients (B/Cd plants) or deprived of potassium ((B-K)/Cd) or calcium ((B-Ca)/Cd) or nitrogen ((B-N)/Cd). Using this approach, we demonstrated that K+ and Ca2+ uptake was impaired in roots exposed to Cd2+. Concerning N, we noticed no indication of uptake inhibition by Cd2+. However, restriction of K+ uptake by roots was compensated by an increase in the K+-use efficiency, so that growth was not inhibited. Calcium uptake was strongly limited by Cd2. This inhibition was accompanied by a reduction in growth of ((B-Ca)/Cd) plants. Thus, we conclude that Cd2+ limits growth of both halophytes through restriction imposed on Ca2+ uptake. We suggest that the increase of Ca2+ availability in soils could improve the growth of both species in the presence of Cd2+. This would be essential for improving their utility for extraction of this metal by from salty contaminated soils. PMID- 17126879 TI - Plasma concentration of organochlorine compounds is associated with age and not obesity. AB - It has been suggested that obese individuals, because of an increased dilution space (body fat) for lipophilic organochlorines compounds, may have greater levels of toxic pollutants than lean sedentary individuals. It is important to further examine this possibility because of the potential contribution of organochlorine pesticides in the development of Parkinson's disease and other neurological diseases. The aim of this study was to further investigate the relationship between the magnitude of obesity and the plasma concentration of organochlorines for a wide range of BMI (with participants at steady state body weight). Fifty-three individuals were selected on the basis of their body mass index (BMI): lean controls (n=16; mean BMI 22.8+/-2.2 kg/m(2); mean age 38.8+/ 9.4 years), obese individuals (n=19; mean BMI 33.4+/-3.0 kg/m(2); mean age 38.6+/ 7.6 years) and morbidly obese individuals (n=18; mean BMI 49.3+/-6.5 kg/m(2); mean age 44.3+/-9.2 years). Blood samples were analyzed for organochlorine compounds. The relationship between the total plasma organochlorine concentration and BMI was tested using a multiple regression analysis. Age was included in the model. There was no relationship between the total plasma organochlorine concentration and BMI. Organochlorine concentrations, however, were correlated with age (BMI-adjusted R(2)=0.46; p<0.001). At steady state body weight, toxic pollutant concentrations are not associated to obesity but strongly correlate with age. PMID- 17126880 TI - Effects of injected activated carbon and solidification treatment on the leachability of polychlorinated dibenzo-p-dioxins and dibenzofurans from air pollution control residues of municipal waste incineration. AB - To assess the effectiveness of the injected activated carbon, cement, and sulfur containing chelating agent in controlling polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) release from the surface of air pollution control (APC) residues, this study examined the leachability of PCDD/Fs from APC residues generated by municipal solid waste incinerators. Results showed that PCDD/Fs were stably retained in the APC residues when the samples were leached with acetic acid solution. Highly chlorinated PCDD/F homologues (i.e., hepta- and octa-CDDs and CDFs) were relatively easy to leach. The leaching percentages of PCDD/Fs from raw APC residue samples containing activated carbon were smaller than those from samples without activated carbon, especially when n-hexane was used as the leachant. These results indicate that the flue gas injected activated carbon not only controls PCDD/F emissions, but also suppresses the leachability of PCDD/Fs from the APC residues. Solidification/stabilization (S/S) processes with 30wt% cement and 5wt% sulfur-containing agent can additionally decrease the leachability of PCDD/Fs with humic acid. Using n-hexane as the leachant, S/S processes increased the leachability of PCDD/Fs. Various low chlorinated PCDD/F congeners were moreover leached out of the APC residue samples, markedly increasing the leachate toxicity. The enhancement of leachability and toxicity owing to S/S processes may negatively impact the environment when APC residues are exposed to nonpolar organic solvents. PMID- 17126881 TI - Rate of hydrolysis and degradation of the cyanogenic glycoside - dhurrin - in soil. AB - Cyanogenic glycosides are common plant toxins. Toxic hydrogen cyanide originating from cyanogenic glycosides may affect soil processes and water quality. In this study, hydrolysis, degradation and sorption of dhurrin (4-hydroxymandelonitrile beta-d-glucoside) produced by sorghum has been studied in order to assess its fate in soil. The log K(ow) of dhurrin was -1.18+/-0.08 (22 degrees C). Hydrolysis was a first-order reaction with respect to dhurrin and hydroxyl ion concentrations. Half lives ranged from 1.2h (pH 8.6; 25 degrees C) to 530d (pH 4; 25 degrees C). The activation energy of hydrolysis was 112+9kJ. At pH 5.8 and room temperature, addition of humic acids (50gl(-1)) increased the rate of hydrolysis tenfold, while addition of kaolinite or goethite (100-250gl(-1)) both decreased the rate considerably. No significant sorption to soil components could be observed. The degradation rates of dhurrin in top and subsoils of Oxisols, Ultisols, Alfisols and Mollisols were studied at 22 degrees C (25mgl(-1), soil:liquid 1:1 (w:V), pH 3.8-8.1). Half-lives were 0.25-2h for topsoils, and 5 288h in subsoils. Hydrolysis in solution explained up to 45% of the degradation in subsoils whereas the contribution in topsoils was less than 14%, indicating the importance of enzymatic degradation processes. The highest risk of dhurrin leaching will take place when the soil is a low activity acid shallow soil with low content of clay minerals, iron oxides and humic acids. PMID- 17126882 TI - Light-induced degradation of perfluorocarboxylic acids in the presence of titanium dioxide. AB - The UV-photon-induced degradation of heptafluorobutanoic acid was investigated in acidic aqueous solutions in the presence of titanium dioxide. Heptafluorobutanoic acid could be degraded with this photocatalyst in a light-induced reaction generating carbon dioxide and fluoride anions. Carbon dioxide evolution in a significant amount occurred only in the presence of molecular oxygen and the photocatalyst. The light-induced degradation of trifluoroacetic acid, pentafluoropropanoic acid, nonafluorobutanoic acid, pentadecafluorooctanoic acid, nonafluorobutanesulfonic acid, and heptadecafluorooctanesulfonic acid in the presence of titanium dioxide was also studied. The perfluorocarboxylic acids under investigation are degraded to generate CO(2) and fluoride anions while both perfluorinated sulfonic acids are persistent under the experimental conditions employed in this study. For all compounds photonic efficiencies of the mineralization reaction were estimated to be smaller than 1x10(-5). To increase the photocatalytic activity mixed systems containing homogeneous phosphotungstic acid and heterogeneous titanium dioxide catalysts were also investigated. In the mixtures of these two photocatalysts, the formation rate of CO(2) increased with illumination time. PMID- 17126883 TI - Monitoring PCDD/Fs and other organic substances in workers of a hazardous waste incinerator: a case study. AB - The aim of this study was to measure, 6 years after regular operations, the concentrations of a number of organic substances in blood and urine of 19 workers employed at a hazardous waste incinerator (HWI) in Spain, and to establish the temporal variation with respect to baseline data and previously performed surveys. This facility was the first, and so far the only HWI in that country. The levels of hexachlorobenzene (HCB), polychlorinated biphenyls (PCB 28, 52, 101, 138, 153 and 180) and polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) were analyzed in six composite plasma samples, while concentrations of di-, tri, and pentachlorophenols, as well those of 1-hydroxypyrene were measured in the urine of these workers. The current mean PCDD/F concentration, 10.4 ng I TEQ/kg lipid, was significantly lower than that found in the baseline survey, 26.7 ng I-TEQ/kg lipid and similar to that found in the previous (2004) study (7.7 ng I-TEQ/kg lipid). PCDD/F levels in plasma were similar or even lower than those recently reported for various non-exposed populations. For the remaining analyzed substances in plasma and urine, there was not any significant increase in comparison with the levels found in the baseline survey. On the other hand, no marked differences between the concentrations of organic substances in plasma or urine were found according to the respective workplace (plant, laboratory and administration). The results of the present survey indicate that after 6 years of regular operation, the workers at the HWI are not occupationally exposed to PCDD/Fs and other organic substances in their workplaces. PMID- 17126884 TI - WO3 thin films for photoelectrochemical purification of water. AB - Tungsten trioxide thin films on transparent substrates (glass and F:SnO(2) or ITO coated glass) were prepared by layer-by-layer brush painting and spin-coating using organic precursors. Well-crystallized WO(3) with monoclinic structure was formed on all substrates after annealing at 500 degrees C or above. The dense semiconducting films are specular and transparent outside the band-gap. Their photoactivity in junctions with aqueous electrolytes extends up to 470 nm, with incident photon to current conversion efficiencies around 0.9 at 313 nm and up to 0.1 at 436 nm. Films of 10 cm x 10 cm were used for the study of solute degradation reactions in a thin-film reactor under backside illumination. Dilute aqueous solutions of model substances for contaminants like oxalic acid were decomposed under continuous flow using broadband UVA illumination and electrical bias. Operation under solar illumination was also feasible. The advantage over operation without bias (conventional photocatalysis) prevailed for all decomposition reactions studied. PMID- 17126885 TI - Degradation mechanisms of benzo[a]pyrene and its accumulated metabolites by biodegradation combined with chemical oxidation. AB - A high degradation extent of benzo[a]pyrene (BaP) should not be considered as the sole desirable criterion for the bioremediation of BaP-contaminated soils because some of its accumulated metabolites still have severe health risks to human. Two main metabolites of BaP, benzo[a]pyrene-1,6-quinone (BP1,6-quinone) and 3 hydroxybenzo[a]pyrene (3-OHBP) were identified by high performance liquid chromatography (HPLC) with standards. This study was the first time that degradation of both BaP and the two metabolites was carried out by chemical oxidation and biodegradation. Three main phases during the whole degradation process were proposed. Hydrogen peroxide-zinc (H(2)O(2)-Zn), the fungus - Aspergillus niger and the bacteria - Zoogloea sp. played an important role in the different phases. The degradation parameters of the system were also optimized, and the results showed that the effect of degradation was the best when fungus bacteria combined with H(2)O(2)-Zn, the concentration range of BaP in the cultures was 30-120mg/l, the initial pH of the cultures was 6.0. However, as co metabolites, phenanthrene significant inhibited the degradation of BaP. This combined degradation system compared with the conventional method of degradation by domestic fungus only, enhanced the degradation extent of BaP by more than 20% on the 12d. The highest accumulation of BP1,6-quinone and 3-OHBP were reduced by nearly 10% in the degradation experiments, which further proved that the combined degradation system was more effective as far as joint toxicity of BaP and its metabolites are concerned. PMID- 17126886 TI - Ozonation of aniline promoted by activated carbon. AB - The removal of aniline from aqueous solutions by simultaneous use of ozone and activated carbon was investigated at different solution pH. For comparative purposes, single ozonation and adsorption on activated carbon were carried out in the same experimental set-up. In order to evaluate the role of the activated carbon surface chemistry during ozonation, a commercial activated carbon, Norit GAC 1240 PLUS, was submitted to oxidation in the liquid phase with HNO(3). The texture and surface chemistry of the activated carbon samples were characterized. During ozonation, complete conversion of aniline was achieved after approximately 20 min, regardless of the presence of activated carbon. In all cases, several by products were formed during ozonation. Nitrobenzene, o- and p-aminophenol were the primary aromatic oxidation by-products identified. In terms of TOC removal, best results were achieved by the simultaneous use of ozone and activated carbon. Though there is a strong contribution of adsorption, a considerable synergetic effect between ozone and activated carbon is observed. In general, activated carbon promotes the reaction of ozonation enhancing the efficiency of this treatment process. The basic activated carbon presented greater activity in this process leading to higher mineralization rates. PMID- 17126887 TI - Protein microarray technology. AB - Protein chips have emerged as a promising approach for a wide variety of applications including the identification of protein-protein interactions, protein-phospholipid interactions, small molecule targets, and substrates of proteins kinases. They can also be used for clinical diagnostics and monitoring disease states. This article reviews current methods in the generation and applications of protein microarrays. PMID- 17126888 TI - Trace element concentrations in the Pacific harbor seal (Phoca vitulina richardii) in central and northern California. AB - To determine concentrations of trace elements (THg, MeHg, Se, and Pb) in tissues of the Pacific harbor seal (Phoca vitulina richardii), live (n=186) and dead seals (n=53) were sampled throughout central and northern California from March 2003 to January 2005. There were significant differences in THg concentrations in blood and hair based on age (p<0.001). Adult male harbor seals had greater THg concentrations in their hair than adult female harbor seals (p<0.003). THg concentrations in liver increased linearly with age and delta15N (p<0.001); whereas, MeHg concentrations in liver increased exponentially until approximately 5 years of age with an asymptote at 1.3 microg/g wet weight. MeHg expressed as a percentage of THg (%MeHg) was best described by a decay function (r2=0.796, p<0.001), decreasing to a minimum at 4 years of age. Hepatic Se increased with age and was in equimolar ratios with THg in adults; whereas, molar ratio of Se:THg in pups deviated from a 1:1 ratio. Significant differences among study locations in THg concentrations in blood and hair were not detected. Assessing the possible effect of sampling location on Hg concentrations, however, was confounded and limited by lack of equal sample sizes for basic age and sex cohorts, a common dilemma in pinniped research. PMID- 17126889 TI - Interrelation of hyperhomocysteinemia and inherited risk factors for venous thromboembolism. Results from the E.D.I.TH. study: a hospital-based case-control study. AB - BACKGROUND: Moderate hyperhomocysteinemia and factor V Leiden mutation are among the most prevalent risk factors for venous thromboembolism (VTE). The hypothesis of an interaction between those risks has been raised and conflicting results were reported. METHODS: We designed a hospital-based case-control study to test the interaction between Factor V Leiden and fasting serum total homocysteine (tHcy). We have also analysed the G20210A prothrombin gene variant. This study enrolled 904 hospitalised patients who had an objectively proven deep vein thrombosis and/or pulmonary embolism as well as 904 hospitalised control patients matched for gender, age and major acquired risk factor for VTE. RESULTS: Our data did not detect any multiplicative interaction between hyperhomocysteinemia (>15 mumol/L) and factor V Leiden mutation or G20210A prothrombin gene variant. Odds ratios (95% CI) were 4.0 (1.5-11) and 6.0 (1.3-27) for the combined effect of hyperhomocysteinemia with either factor V Leiden mutation or G20210A prothrombin gene variant, respectively. CONCLUSIONS: Current data provide further knowledge in relationship between hyperhomocysteinemia and inherited risk factors, such as factor V Leiden mutation and G20210A prothrombin gene variant. As those risk factors are not so rare among Caucasians, a better estimate of the risk related to double exposure might help to optimise venous thromboembolism prevention. PMID- 17126890 TI - Ovarian serous carcinoma presenting with mediastinal lymphadenopathy 20 months before the intraabdominal mass: role of immunohistochemistry. AB - INTRODUCTION: Metastatic involvement of the mediastinal lymph nodes is an extremely uncommon finding in epithelial ovarian cancers. CASE REPORT: A 63-year old woman was admitted to hospital for dyspnoea due to an anterior mediastinal mass. The surgical biopsy showed a 6-cm metastatic lymph node with a papillary pattern, scattered psammomas and immunoreactivity for WT1, Cytokeratin 7, EMA and negative for E-cadherin, GCFDP-15, Thyroglobulin, Cytokeratin 20, Cytokeratin 5/6, CEA, Vimentin, Calretinin, TTF1. After 20 months a follow-up CT identified an ovarian mass with the same histological pattern and immunoreactivity of the mediastinal nodule. RESULTS: Immunohistochemistry, especially WT1, is useful in assessing the ovarian origin of an unusual metastasis particularly if it is the first presentation of the disease. PMID- 17126891 TI - Positron emission tomography and leiomyomas: clinicopathologic analysis of 3 cases of PET scan-positive leiomyomas and literature review. AB - INTRODUCTION: Studies have suggested that PET scans can differentiate between leiomyomas and leiomyosarcomas. Our experience, however, shows that PET scan positive smooth muscle tumors are not necessarily malignant. CASE REPORTS: Three patients with cancer underwent PET imaging. In all three, the most worrisome finding was a PET scan-positive uterine tumor. After surgical extirpation, all three uterine tumors were found to be benign smooth muscle neoplasms. DISCUSSION: To explore the potential reason these tumors were positive on PET imaging, we performed a detailed histopathologic and immunohistochemical study of all specimens. Pathologic evaluation revealed a leiomyoma, a cellular leiomyoma, and a stromomyoma. There was no association between an increased Ki67 (proliferative) index and positivity on PET imaging. Increased vascularity, however, appeared to be a feature common to the leiomyomas that were PET-positive. PMID- 17126892 TI - Endometrial cancer--revisiting the importance of pelvic and para aortic lymph nodes. AB - In 1998, FIGO (International Federation of Gynecologists and Obstetricians) required a change from clinical to surgical staging in early endometrial cancer. This staging requirement raised numerous controversies around the importance of determining nodal status and its impact on outcomes. A diversity of opinions exists as to the actual benefits and toxicities associated with surgical staging which includes lymph node sampling, ranging from those whose opinion is that staging is required for all patients even when the a priori risk of nodal involvement is extremely low through to those who consider that staging is unnecessary in any patient. While knowledge of the presence or absence of extra uterine sites of disease may change treatment approaches and direct different treatment interventions in some patients, the impact of those changes on survival is much less clear. This paper examines recommendations for surgical staging in various subgroups of patients with clinically early endometrial cancer and the impact on survival and toxicity of the various approaches and the subsequent use of adjuvant irradiation and/or chemotherapy. PMID- 17126893 TI - Assessment of Her-2/neu expression in hydatidiform moles for prediction of subsequent gestational trophoblastic neoplasia. AB - OBJECTIVE: The aim of the present study was to asses the ability of Her-2/neu immunohistochemical staining of the molar tissue to predict the risk of developing gestational trophoblastic neoplasia (GTN). METHODS: Sections prepared from 33 consecutive formalin-fixed paraffin-embedded archival reconfirmed hydatidiform mole tissue blocks were immunohistochemically stained for Her-2/neu. The staining was scored according to the subjectively evaluated intensity of staining and the proportion of stained villous cytotrophoblastic cells. Clinical data were abstracted from medical files. RESULTS: 23 patients had a complete and 10 a partial mole. Nine patients (27.3%) were diagnosed with GTN [7 of 23 patients with a complete mole (30.4%) and 2 of the 10 (20.0%) with a partial mole]. A positive immunohistochemical Her-2/neu stain was found in 6 (18.2%) of the patients with hydatidiform mole (3 with a complete mole). The rate of Her 2/neu expression was somewhat higher in moles with subsequent GTN than in moles with an uneventful course (22.2% vs. 16.6%, respectively). The difference did not reach significance (Fisher's Exact Test, P=0.55) possibly due to the small number of cases (power of <5%). The sensitivity and specificity of Her-2/neu expression for prediction of GTN was 22.2% and 83.3%, respectively, and the positive and negative predictive value 33.3% and 74.1%, respectively. CONCLUSION: While the specificity of Her-2/neu immunohistochemical staining for prediction of GTN is relatively high, the low sensitivity and low positive and negative predictive value precludes its practical clinical use for prediction of post-molar GTN. The quest for a precise predictor of post-molar GTN should continue. PMID- 17126894 TI - A phase II trial of EMD72000 (matuzumab), a humanized anti-EGFR monoclonal antibody, in patients with platinum-resistant ovarian and primary peritoneal malignancies. AB - OBJECTIVE: The primary objective of this study was to determine the rate of response to matuzumab in patients with recurrent, EGFR-positive ovarian, or primary peritoneal cancer. Secondary end points included safety and tolerability, time to tumor progression, duration of response, and overall survival. METHODS: A multi-institutional single arm phase II trial. RESULTS: Of 75 women screened for the study, 37 were enrolled and treated. Median age of the treated patient population was 58 years, and most patients had more than four prior lines of chemotherapy. Therapy was well tolerated, the most common toxicities being a constellation of skin toxicities, including rash, acne, dry skin, and paronychia, as well as headache, fatigue, and diarrhea. Serious adverse events were very rare but included a single episode of pancreatitis that may have been drug related. All patients completed therapy, receiving 1 to 30 infusions of matuzumab. There were no formal responses (RR=0%, 95% CI: 0-9.5%), although 7 patients (21%) were on therapy for more than 3 months with stable disease. CONCLUSIONS: Matuzumab at the dose and schedule selected is well tolerated. In this population of very heavily pretreated patients with epithelial ovarian and primary peritoneal malignancies, there was no evidence of significant clinical activity when matuzumab was administered as monotherapy. PMID- 17126895 TI - Intrauterine Neospora caninum inoculation of heifers and cows using contaminated semen with different numbers of tachyzoites. AB - OBJECTIVE: To investigate the potential of different Neospora caninum tachyzoite doses to infect heifers (experiment 1) and cows (experiment 2) when administered in utero by artificial insemination via contaminated semen. METHODS: In experiment 1, five groups of 5, 7, 8, 9, and 5 cyclic heifers were hormonally synchronized and artificially inseminated with semen containing 0 (A, controls), 10(2) (B), 5 x 10(3) (C), 5 x 10(4) (D), and 5 x 10(5) (E) live N. caninum NC-1 isolate-tachyzoites, respectively. Experimental infection was followed for 100 days. Parasitaemia and specific serum IgG, and interferon-gamma (IFN-gamma) responses were studied. In experiment 2, four groups of 9, 10, 9, and 9 adult multiparous cows with confirmed infertility problems of diverse aethiology were hormonally synchronized and artificially inseminated with semen containing 0 (a, controls), 10(2) (b), 5 x 10(3) (c), and 5 x 10(5) (d) live N. caninum NC-1 isolate-tachyzoites, respectively. Experimental infection was followed for 63 days. Parasitaemia and specific serum IgG responses were studied. RESULTS: In experiment 1, parasitaemia was detected in 1, 2, and 3 heifers from groups B, C, and D, respectively, between 9 and 23 days after insemination. Persistent specific serum antibody responses were detected in 2 and 3 heifers from groups D and E, respectively. Transient specific serum antibody responses were detected in 2, 1 and 1 heifers from groups C, D, and E, respectively. In addition, 1 heifer from group B showed a serum-specific antibody level higher than cut off value at 21 days post-insemination. Heifers seroconverted between 23 and 47 days after insemination. Specific IFN-gamma levels were detected in 1, 4, 6, and 3 heifers from groups B, C, D, and E, respectively, between 9 and 55 days after insemination. Pregnancy rate in the control group (60%) was higher than those observed in inoculated heifers (0-42.9%). Pregnancy rates in inoculated heifers were lower when the tachyzoite dose was increased (B 42.9%, C 12.5%, D 11.1%, and E 0%). In experiment 2, no Neospora DNA in blood nor specific serum IgG to N. caninum were detected in any of the cows studied, except in one cow inoculated with 5 x 10(5) tachyzoites (group d) which showed a relative index x100 (RIPC) values of 9.4, 18.9, and 18.1 at 42, 56, and 63 days after insemination, respectively. CONCLUSIONS: This study provides evidence that the intrauterine infection via contaminated semen using 5 x 10(4) and 5 x 10(5) tachyzoites caused persistent serum-specific antibody responses in some heifers. On the basis of serological data, a dose-response effect was also observed. In addition, N. caninum would be a probable cause of early foetal death in inoculated heifers. In contrast, results obtained in a similar experiment with cows showing confirmed infertility indicate that higher doses, such as of 5 x 10(5) tachyzoites, were necessary to induce seroconversion in at least one animal. PMID- 17126896 TI - The effects of the prostaglandin E analogue Misoprostol and follicle-stimulating hormone on cervical penetrability in ewes during the peri-ovulatory period. AB - Two experiments in parous Welsh Mountain ewes determined the pattern of natural cervical relaxation over the peri-ovulatory period and investigated FSH and Misoprostol as cervical relaxants to facilitate transcervical passage of an insemination pipette into the uterine cavity. Following synchronisation of oestrus using progestagen sponges and PMSG (500 IU) the depth of cervical penetration was determined using a modified cattle insemination pipette as a measuring device. Penetration of the cervix was least at the time of sponge removal and increased to a maximum at 72 h after sponge removal and then declined. Intra-cervical administrations of either ovine FSH (Ovagen; 2mg) or Misoprostol (1mg; a Prostaglandin E(1) analogue) facilitated cervical penetration. Ovagen given 24h after sponge removal allowed transcervical intrauterine penetration in 100% of ewes at 54 and 60 h after sponge removal while Misoprostol given 48 h after sponge removal allowed trans-cervical penetration in 100% of ewes at 54 h. A combination of Ovagen and Misoprostol was as effective but not more so than Ovagen or Misoprostol alone. These results show that there is natural relaxation of the cervix at oestrus and that maximum relaxation occurs 72 h after sponge removal, which is too late for the correct timing of insemination. The intra-cervical administration of FSH or Misoprostol enhanced relaxation of the cervix and both were able to relax the cervix to allow intrauterine penetration 54 h after sponge removal, the optimum time for insemination. The results also show that FSH is biologically active after intracervical, topical application. PMID- 17126897 TI - Microscopic and flow cytometric semen assessment of Dutch AI-bucks: effect of semen processing procedures and their correlation to fertility. AB - This study was done to determine the effects of processing techniques on the quality of semen from Dutch AI-bucks with the view on improving pregnancy rates after artificial insemination (AI) with liquid or frozen-thawed semen. Motility of spermatozoa was estimated under a microscope whereas the percentage live spermatozoa and the percentage live spermatozoa with intact acrosomes were determined by means of flow cytometry. Aspects of semen processing that were investigated are storage temperature of liquid semen (i), the effect of glycerol on liquid-stored semen (ii), removal of seminal plasma (iii) and type of extender (iv). The correlation between semen quality and fertility rates in inseminated does was also investigated. The percentage motile spermatozoa in semen stored in liquid form for 72 h progressively declined over time, irrespective of whether storage occurred at 4 or 18 degrees C. The percentage motile spermatozoa in semen stored at 18 degrees C was similar to that in semen stored at 4 degrees C if stored for 24 h but lower if stored for 48 h. Goats differ in the sensitivity of their spermatozoa to the deleterious effects of glycerol. Neither the removal of seminal plasma nor the type of extender had any effect on semen quality before freezing but semen frozen in a Tris-citric acid-glucose (TCG) buffer with egg yolk without removal of the seminal plasma had better quality after thawing than semen frozen in another diluent or after removal of seminal plasma. Remarkably no significant correlation between fertility and membrane integrity of spermatozoa could be found. Thus, although integrity assays for spermatozoa are useful to asses resistance to semen handling, the validity of these assays for predicting fertility is questioned. PMID- 17126898 TI - Preparation of hemoglobin-loaded nano-sized particles with porous structure as oxygen carriers. AB - Hb (hemoglobin)-loaded particles (HbP) encapsulated by a biodegradable polymer used as oxygen carrier were prepared. A modified double emulsion and solvent diffusion/evaporation method was adopted. All experiments were performed based on two types of biodegradable polymers, poly(epsilon-caprolactone) (PCL) and poly(epsilon-caprolactone-ethylene glycol) (PCL-PEG). The biodistribution and the survival time in blood of the particles were investigated in a mouse model. Encapsulation efficiency and pore-connecting efficiency were evaluated by a novel sulfocyanate potassium method. The influence of process parameters on the particle size and pore-connecting efficiency (PCE%) of nanoparticles have been discussed. The prepared conditions: solvent, external aqueous phase, pressure were discussed. The system utilizing dichloromethane (DCM)/ethyl acetate (EA) as a solvent with an unsaturated external aqueous phase yielded the highest encapsulation efficiency (87.35%) with a small mean particle size (153 nm). The formation of porous channels was attributed to the diffusion of solvent. The PCE% was more sensitive to the rate of solvent diffusion that was obviously affected by the preparation temperature. The PCE% reached 87.47% when PCL-PEG was employed at 25 degrees C. P(50) of HbP was 27 mmHg, which does not seem to be greatly affected by the encapsulation procedure. In vivo, following intravenous injection of 6-coumarin labeled HbP, the major organ accumulating Hb-loaded particles was the liver. The half-life of nano-sized PCL HbP was 3.1 times as long as the micro sized PCL HbP. Also, Nano-sized as well as a PEGylated surface on HbP is beneficial for prolonged blood residence (7.2 fold increase). PMID- 17126899 TI - The inhibition of TNF-alpha-induced E-selectin expression in endothelial cells via the JNK/NF-kappaB pathways by highly N-acetylated chitooligosaccharides. AB - Chitooligosaccharides (COS) have been shown to regulate various cellular and biological functions. However, the effect of COS on inflammatory responses of the cells remains unclear. We investigated the regulatory effect of highly N acetylated COS (NACOS) on tumor necrosis factor-alpha (TNF-alpha)-induced endothelial cell (EC) E-selectin expression, which is crucial for leukocyte recruitment. ECs were kept as controls or pre-treated with NACOS for different times, and then stimulated with TNF-alpha for 4h. The results show that pre treating ECs with NACOS inhibited the TNF-alpha-induced E-selectin expression in a dose- and time-dependent manner. This NACOS-mediated inhibition in E-selectin expression was regulated at the transcriptional level, but not due to changes in mRNA stability. Stimulation of ECs with TNF-alpha-induced rapid increases in the phosphorylation of their mitogen-activated protein kinases (MAPKs) [extracellular signal-regulated kinase (ERK), c-Jun-NH2-terminal kinase (JNK), and p38 MAPK]; the inhibitor for JNK (i.e., SP600125), but not those for ERK (i.e., PD98059) and p38 MAPK (i.e., SB203580), attenuated this TNF-alpha-induced E-selectin expression. Pre-treating ECs with NACOS inhibited the TNF-alpha-induced JNK activation, suggesting that JNK was involved in the inhibitory effect of NACOS on TNF-alpha-induced E-selectin expression. Pre-treating ECs with NACOS inhibited the TNF-alpha-induced p65 and p50 mRNA expressions. Gel shifting and chromatin immunoprecipitation assays showed that NACOS blocked the TNF-alpha-induced increases in the binding activity and in vivo promoter binding of nuclear factor kappaB (NF-kappaB) in ECs. Our findings provide a molecular mechanism by which NACOS inhibit TNF-alpha-induced E-selectin expression in ECs, and a basis for using NACOS in pharmaceutical therapy against inflammation. PMID- 17126900 TI - In vivo tumor targeting and radionuclide imaging with self-assembled nanoparticles: mechanisms, key factors, and their implications. AB - The development of more selective delivery systems for cancer diagnosis and chemotherapy is one of the most important goals of current anticancer research. The purpose of this study is to evaluate various self-assembled nanoparticles as candidates to shuttle radionuclide and/or drugs into tumors and to investigate the mechanisms underlying the tumor targeting with self-assembled nanoparticles. By combining different hydrophobic moieties and hydrophilic polymer backbones, various self-assembled nanoparticles were prepared, and their in vivo distributions in tumor-bearing mice were studied by radionuclide imaging. One type of nanoparticles (fluorescein isothiocyanate-conjugated glycol chitosan (FGC) nanoparticles) exhibited highly selective tumoral localization. Scintigraphic images obtained 1 day after the intravenous injection of FGC nanoparticles clearly delineated the tumor against adjacent tissues. The mechanisms underlying the tumor targeting with self-assembled nanoparticles were investigated in terms of the physicochemical properties of nanoparticles and tumor microenvironments. FGC nanoparticles were preferentially localized in perivascular regions, implying their extravasation to tumors through the hyperpermeable tumor vasculature. The magnitude and pattern of tumoral distribution of self-assembled nanoparticles were influenced by several key factors--(i) in vivo colloidal stability: nanoparticles should maintain their intact nanostructures in vivo for a long period of time, (ii) particle size, (iii) intracellular uptake of nanoparticle: fast cellular uptake greatly facilitates the tumor targeting, (iv) tumor angiogenesis: pathological angiogenesis permits access of nanoparticles to tumors. We believe that this work can provide insight for the engineering of nanoparticles and be extended to cancer therapy and diagnosis, so as to deliver multiple therapeutic agents and imaging probes at high local concentrations. PMID- 17126901 TI - The influence of polymeric properties on chitosan/siRNA nanoparticle formulation and gene silencing. AB - We have previously introduced the use of the biomaterial chitosan to form chitosan/siRNA nanoparticles for gene silencing protocols. This present study shows that the physicochemical properties (size, zeta potential, morphology and complex stability) and in vitro gene silencing of chitosan/siRNA nanoparticles are strongly dependent on chitosan molecular weight (Mw) and degree of deacetylation (DD). High Mw and DD chitosan resulted in the formation of discrete stable nanoparticles approximately 200 nm in size. Chitosan/siRNA formulations (N:P 50) prepared with low Mw (approximately 10 kDa) showed almost no knockdown of endogenous enhanced green fluorescent protein (EGFP) in H1299 human lung carcinoma cells, whereas those prepared from higher Mw (64.8-170 kDa) and DD (approximately 80%) showed greater gene silencing ranging between 45% and 65%. The highest gene silencing efficiency (80%) was achieved using chitosan/siRNA nanoparticles at N:P 150 using higher Mw (114 and 170 kDa) and DD (84%) that correlated with formation of stable nanoparticles of approximately 200 nm. In conclusion, this work confirms the application of chitosan as a non-viral carrier for siRNA and the importance of polymeric properties for the optimisation of gene silencing using chitosan/siRNA nanoparticles. PMID- 17126902 TI - DNA vaccine-encoded glycoprotein B of HSV-1 fails to protect chronic morphine treated mice against HSV-1 challenge. AB - The use of morphine has been demonstrated to increase susceptibility to infections. Herpes simplex virus type 1 (HSV-1) is a highly successful pathogen among immunocompromised individuals. In the present study, due to the importance of HSV vaccination in morphine abusers, the effects of chronic morphine exposure on the host response to a HSV-1 gB DNA-based vaccine have been investigated. The study is addressing an important aspect of vaccine development among the susceptible (immunocompromised) hosts. BALB/c mice were exposed to morphine over 11 days. They were then vaccinated with DNA vaccine or KOS strain as a live vaccine. The findings showed that the morphine-treated animals failed to respond to DNA vaccination evaluated by the anti-HSV gB antibody titer, delayed type hypersensitivity (DTH) and lethal HSV-1 challenge. Under the same conditions, the KOS vaccine showed a reduced Ab titer and DTH response in morphine-treated mice, but could protect mice against the lethal challenge and was safe for vaccination of morphine-treated animals. PMID- 17126903 TI - The genetic background to PTSD. AB - Although extensive research has already been done on the genetic bases of psychiatric disorders, little is known about polygenetic influences in posttraumatic stress disorder (PTSD). This article reviews molecular genetic studies relating to PTSD that were found in a literature search in Medline, Embase and Web of Science. Association studies have investigated 8 major genotypes in connection with PTSD. They have tested hypotheses involving key candidate genes in the serotonin (5-HTT), dopamine (DRD2, DAT), glucocorticoid (GR), GABA (GABRB), apolipoprotein systems (APOE2), brain-derived neurotrophic factor (BDNF) and neuropeptide Y (NPY). The studies have produced inconsistent results, many of which may be attributable to methodological shortcomings and insufficient statistical power. The complex aetiology of PTSD, for which experiencing a traumatic event forms a necessary condition, makes it difficult to identify specific genes that substantially contribute to the disorder. Gene finding strategies are difficult to apply. Interactions between different genes and between them and the environment probably make certain people vulnerable to developing PTSD. Gene-environmental studies are needed that focus more narrowly on specific, distinct endophenotypes and on influences from environmental factors. PMID- 17126904 TI - Functional region of mouse heat shock protein 72 for its binding to lymphoid neoplastic P388D1 cells. AB - Extracellular heat shock proteins have been reported to participate in both innate and adaptive immune responses. We have found that recombinant mouse inducible heat shock protein 72 (Hsp72) bound to lymphoid neoplastic P388D1 cells. In the present study, we examined which region of mouse Hsp72 interacted with this cell line by using truncated variants that are sequentially lacking sections of the C-terminal region. The full-length mouse Hsp72 specifically bound to P388D1 cells, but not to mastocytoma P815 cells. Deletion of the C-terminal tail portion of mouse Hsp72 markedly decreased the binding to P388D1 cells and the sequential truncation of the C-terminal helical region led to a loss of binding activity. Specific binding was not observed for either the variant with a minimal substrate-binding structure or the ATP-binding domain alone. On the other hand, two truncated variants lacking the ATP-binding domain significantly bound to P388D1 cells. However, the variant lacking the substrate-binding domain did not show any binding to this cell line. These results suggest that the activity to bind P388D1 cells is attributable to the C-terminal region of mouse Hsp72 in combination with the substrate-binding domain. Interestingly, the binding of mouse Hsp72 to P388D1 cells was competed by the variant with the C-terminal flexible tail sequence, but not by the variant without that sequence. These competitive experiments imply that there may be at least two membrane receptors on P388D1 cells and also that both receptors may recognize the various structures in the C-terminal region of the Hsp70 family for regulation of innate immunity. PMID- 17126905 TI - Differential regulation of TNFalpha and GM-CSF induced activation of P38 MAPK in neutrophils and eosinophils. AB - P38 MAPK is a central mediator in cytokine signalling in human leukocytes. P38 MAPK is activated by phosphorylation of a conserved Thr180-X-Tyr182 motif by dual phosphorylation via the upstream kinases MKK3 and MKK6. Alternatively, P38 MAPK can be activated via autophosphorylation when associated with TAB1. In this study P38 MAPK phosphorylation and activation (measured via phosphorylation of P38 MAPK downstream target MK2) were investigated upon engagement of the GM-CSF- and TNFalpha-receptors expressed on both eosinophils and neutrophils. The MKK3/MKK6 pathway mediated neutrophil P38 MAPK activation after stimulation with TNFalpha (100U/ml) or GM-CSF (10(-10)M). Under these conditions the activation but not phosphorylation of P38 MAPK could be inhibited by SB203580 (10(-5)M or 10(-6)M). In eosinophils SB203580 (10(-6)M) inhibited both the phosphorylation and activation of P38 MAPK after stimulation with several doses of TNFalpha (10-10000 U/ml) or GM-CSF (10(-11) to 10(-9)M), indicating that P38 MAPK activation is mediated via autophosphorylation in eosinophils. This hypothesis was supported by the finding that in marked contrast to neutrophils, MKK3/MKK6 did not show enhanced phosphorylation in eosinophils after cytokine stimulation, but were constitutively phosphorylated. Therefore, the involvement of TAB1 was investigated with regard to this cytokine-induced autophosphorylation. Co immunoprecipitation experiments showed that TAB1 was constitutively associated with P38 MAPK in eosinophils and neutrophils and that cytokine-induced autophosphorylated P38 MAPK was co-precipitated with TAB1. These findings are consistent with the hypothesis that cytokine-induced autophosphorylation of P38 MAPK in primary granulocytes depends on the interaction with TAB1. PMID- 17126906 TI - DNA-binding and cleavage studies of novel binuclear copper(II) complex with 1,1' dimethyl-2,2'-biimidazole ligand. AB - DNA-binding properties of novel binulear copper(II) complex [Cu(2)(Dmbiim)(4)(H(2)O)(2)](ClO(4))(4).6H(2)O, where Dmbiim = 1,1'-Dimethyl-2,2' biimidazole are investigated using electronic absorption spectroscopy, fluorescence spectroscopy, viscosity measurement and voltammetry. The results show that the copper(II) complex interacts with DNA through minor groove binding. The interaction between the complex and DNA has also been investigated by gel electrophoresis, interestingly, we found that the copper(II) complex can cleave circular plasmid pBR322 DNA efficiently in the presence of AH(2) (ascorbic acid) at pH 8.0 and 37 degrees C. PMID- 17126907 TI - Ni K-edge XAS suggests that coordination of Ni(II) to the unstructured amyloidogenic region of the human prion protein produces a Ni(2) bis-mu-hydroxo dimer. AB - Prion diseases are thought to be caused by the misfolding of the ubiquitous neuronal membrane prion protein (PrP) through an unknown mechanism that may involve Cu(II) coordination to the PrP. Previous work has utilized Ni(II) as a diamagnetic probe for Cu(II) coordination [C.E. Jones, M. Klewpatinond, S.R. Abdelraheim, D.R. Brown, J.H. Viles, J. Mol. Biol. 346 (2005) 1393-1407]. Herein we investigate Ni(II) coordination to the PrP fragment PrP(93-114) (AcN GGTHSQWNKPSKPKTNMKHMAG) at pH=10.0 by Ni K-edge X-ray absorption spectroscopy (XAS). We find that two equivalents of Ni(II) will coordinate to PrP(93-114) by UV/Vis titrations and mass spectrometry. Ni K-edge XAS data is consistent with Ni(II) ligated by five N/O based ligands (three N/O ligands at 2.01(2) Angstrom and two at 1.855(2) Angstrom). We were also able to locate a Ni-Ni vector at 3.1(1) Angstrom, which suggests the two Ni(II) centers are contained in a bis-mu hydroxo dimer. We therefore suggest that Ni(II) may not be a suitable diamagnetic mimic for Cu(II) coordination within the PrP since differential coordination modes for the two metals exist. PMID- 17126908 TI - Matrix solid-phase dispersion as a valuable tool for extracting contaminants from foodstuffs. AB - This review updates our knowledge on matrix solid-phase dispersion (MSPD), a sample treatment procedure that is increasingly used for extracting/purifying contaminants from a variety of solid, semi-solid, viscous, and liquid foodstuffs. MSPD is primarily used because of its flexibility, selectivity, and the possibility of performing extraction and cleanup in one step, this resulting in drastically shortening of the analysis time and low consumption of toxic and expensive solvents. Technical developments and parameters influencing the extraction yield and selectivity are examined and discussed. Experimental results for the analysis of pesticides, veterinary drugs, persistent environmental chemicals, naturally occurring toxicants, and surfactants in food are reviewed. PMID- 17126909 TI - Head-mounted goggles for murine form deprivation myopia. AB - Recently a murine model has been developed for use in form deprivation myopia experiments. Due to the small size of the head and eye, methods to blur visual input to the mouse eye are challenging. Previous methods to induce form deprivation include lid suture and gluing diffuser goggles directly to the fur around the eye. In this paper we describe a new method of goggling using a head pedestal and goggle, which improves compliance and allows for better ocular health. Nob mice, previously shown to be highly susceptible to form deprivation myopia, were used for these experiments. Immediately following baseline refraction using an infrared automated photorefractor, mice were either goggled with a diffuser attached directly to the fur or with a head-mounted goggling apparatus. The goggle apparatus consists of five main components: goggle and frame, head pedestal, acrylic cube for stabilization, and balancing bar. Mice were goggled for 2 weeks in which ocular health and goggle position was monitored and then had a final refraction. The use of head-mounted goggles resulted in 75% fewer instances of goggle loss and 55% fewer ocular complications compared to goggles glued to the fur. Both goggling methods induced a myopic shift of approximately 5 diopters. The head-mounted goggle apparatus provides an improved method for inducing form deprivation in mice and offers the ability to easily take repeated refractive measurements as well as allowing for the use of defocusing lenses. PMID- 17126910 TI - Amygdala activation at 3T in response to human and avatar facial expressions of emotions. AB - Facial expressions of emotions are important in nonverbal communication. Although numerous neural structures have been identified to be involved in emotional face processing, the amygdala is thought to be a core moderator. While previous studies have relied on facial images of humans, the present study is concerned with the effect of computer-generated (avatar) emotional faces on amygdala activation. Moreover, elicited activation patterns in response to viewing avatar faces are compared with the neuronal responses to human facial expressions of emotions. Twelve healthy subjects (five females) performed facial emotion recognition tasks with optimized 3T event-related fMRI. Robust amygdala activation was apparent in response to both human and avatar emotional faces, but the response was significantly stronger to human faces in face-sensitive structures, i.e. fusiform gyri. We suggest that avatars could be a useful tool in neuroimaging studies of facial expression processing because they elicit amygdala activation similarly to human faces, yet have the advantage of being highly manipulable and fully controllable. However, the finding of differences between human and avatar faces in face-sensitive regions indicates the presence of mechanisms by which human brains can differentiate between them. This mechanism merits further investigation. PMID- 17126911 TI - Three fluorescent protein voltage sensors exhibit low plasma membrane expression in mammalian cells. AB - Three first-generation fluorescent protein voltage sensitive probes (FP-voltage sensors) were characterized in mammalian cells. Flare, a Kv1.4 variant of FlaSh [Siegel MS, Isacoff EY. Neuron 1997;19(October (4)):735-41], SPARC [Ataka K, Pieribone VA. Biophys J 2002;82(January (1 Pt 1)):509-16], and VSFP-1 [Sakai R, Repunte-Canonigo V, Raj CD, Knopfel T. Eur J Neurosci 2001;13(June (12)):2314-18] were expressed, imaged and voltage clamped in HEK 293 cells and in dissociated hippocampal neurons. We were unable to detect a signal in response to changes in membrane potential after averaging16 trials with any of the three constructs. Using the hydrophobic voltage sensitive dye, di8-ANEPPS, as a surface marker, confocal analyses demonstrated poor plasma membrane expression for Flare, SPARC and VSFP-1 in both HEK 293 cells and dissociated hippocampal neurons. Almost all of the expressed FP-voltage sensors reside in internal membranes in both cell types. This internal expression generates a background fluorescence that increases the noise in the optical measurement. PMID- 17126912 TI - A comparison of the familiality of chronic depression in recurrent early-onset depression pedigrees using different definitions of chronicity. AB - BACKGROUND: The study of chronicity in the course of major depression has been complicated by varying definitions of this illness feature. Because familial clustering is one component of diagnostic validity we compared family clustering of chronicity as defined in the DSM-IV to that of chronicity determined by an assessment of lifetime course of depressive illness. METHODS: In 1750 affected subjects from 652 families recruited for a genetic study of recurrent, early onset depression, we applied several definitions of chronicity. Odds ratios were determined for the likelihood of chronicity in a proband predicting chronicity in an affected relative. RESULTS: There was greater family clustering of chronicity as determined by assessment of lifetime course (OR=2.54) than by DSM-IV defined chronic major depressive episode (MDE) (OR=1.93) or dysthymic disorder (OR=1.76). In families with probands who had preadolescent onset of MDD, familiality was increased by all definitions, with a much larger increase observed for chronicity by lifetime course (ORs were 6.14 for lifetime chronicity, 2.43 for chronic MDE, and 3.42 for comorbid dysthymic disorder). Agreement between these definitions of chronicity was only fair. LIMITATIONS: The data used to determine chronicity were collected retrospectively and not blindly to relatives' status, and assessment of lifetime course was based on global clinical impressions gathered during a semi structured diagnostic interview. Also, it can be difficult to determine whether individuals with recurrent major depressive episodes who frequently experience long periods of low grade depressive symptoms meet the strict timing requirements of DSM-IV dysthymic disorder. CONCLUSIONS: An assessment of lifetime symptom course identifies a more familial, and thus possibly a more valid, type of chronic depression than the current DSM-IV categories which are defined in terms of particular cross-sectional features of illness. PMID- 17126913 TI - Subsyndromal depression: an impact on quality of life? AB - OBJECTIVE: The objective of this study was to demonstrate the association between quality of life and subsyndromal depression in a primary care clinic in a Brazilian sample. METHODS: This was a cross-sectional study. The cases were divided into three groups according to the severity of depressive symptoms: 1) subjects with major depressive disorder; 2) subjects with subsyndromal depression; 3) subjects without depressive symptoms--controls. The participants completed the World Health Organization Instrument to Assess Quality of Life (WHOQOL-BREF), the Quality of Life--Depression (QLDS), the Centers for Epidemiologic Studies--Depression instrument (CES-D), and the Composite International Diagnostic Interview (CIDI). RESULTS: The sample consisted of 438 primary care users (35.2% of them had subsyndromal depression). The subjects with major depression presented the worst impairment of quality of life, which was measured by the WHOQOL-BREF and the QLDS. The patients with subsyndromal depression had a smaller impact on their quality of life and the subjects without depression presented an even lower impact. The hierarchical linear regression involving demographic variables and the severity of depressive symptoms showed that the severity of depression was the variable with higher correlation with quality of life dimensions, presenting increased variation in the domains (from 9% to 24%). CONCLUSIONS: The results suggest that subsyndromal depression causes impairment of the quality of life in primary care patients of a Brazilian sample. PMID- 17126914 TI - Elevated pain thresholds correlate with dissociation and aversive arousal in patients with borderline personality disorder. AB - In addition to reduced pain perception, patients with borderline personality disorder (BPD) show higher pain thresholds under subjective stress conditions as compared with non-stress conditions. However, the correlation between symptoms of stress and pain thresholds has not been investigated so far. Using a new and convenient methodology, electric stimulation, we expected higher pain and detection thresholds in patients with BPD than in to healthy controls as well as a positive correlation between pain thresholds and symptoms of stress (aversive arousal and dissociation) in BPD patients. Twelve female patients with BPD and twelve healthy controls were included in the study. Electric stimulation was applied on the right index finger, and detection and pain thresholds were assessed by gradually intensifying the stimuli. We found significantly elevated pain thresholds in patients with BPD as compared with healthy controls, but no difference between patients and controls in detection thresholds. In patients, a significant positive correlation was revealed between pain thresholds and dissociation as well as between pain thresholds and aversive arousal. Besides demonstrating a close correlation between pain thresholds and symptoms of stress in patients with BPD, this study replicated earlier findings of reduced pain perception in patients with BPD. Measuring electric pain thresholds is a valid and reasonable method for larger studies. PMID- 17126917 TI - The extent and levels of tinnitus in children of central Ankara. AB - OBJECTIVE: The objective of this study is to determine the presence and prevalence of tinnitus among primary school and junior high school students in central Ankara. METHODS: In the first stage of the study, all students were tested for the presence of tinnitus by answering a comprehensive questionnaire. The students who had previous ear operations were excluded from the rest of the study. The initial survey/tests yielded presence of tinnitus, frequency of occurrence, characteristics, associated symptoms and the age groups. RESULTS: 15.1% of the children reported to have tinnitus. No significant difference was found between gender (female 45.5%, male 54.4%) and ears (right 25.3%, left 25.5%). The age group that suffered most from tinnitus is 14 years old (20.8%), 25 children had positive family history (16.2%), 44 children had headaches as the most common accompanying symptom (28.6%), 64 of them had tiredness as the predisposing factor (41.6%) and 52 of them have defined worsening of tinnitus during mornings (33.8%). The characteristics of tinnitus were identified as high pitch (n=125, 81.2%), soft loudness (n=124, 80.5%) and ringing (n=61, 39.6%). CONCLUSIONS: The study produced much needed data to shed light onto understanding levels and characteristics of tinnitus in school children in Turkey. The data obtained was carefully analyzed and found to be comparative to international studies. PMID- 17126916 TI - The human CMV-UL86 peptide 981-1003 shares a crossreactive T-cell epitope with the encephalitogenic MOG peptide 34-56, but lacks the capacity to induce EAE in rhesus monkeys. AB - Rhesus monkeys immunized with MOG(34-56), a dominant T-cell epitope from myelin/oligodendrocyte glycoprotein, develop an acute neurological disease resembling acute disseminated encephalomyelitis (ADEM) in humans. The typical large demyelinated lesions and mononuclear infiltrates in the monkey brains are caused by MOG(34-56) T-cells. We show that MOG(34-56)-reactive CD4+ and CD8+ T cells are induced in monkeys immunized with a peptide from the human CMV major capsid protein (UL86; 981-1003), that shares sequence similarity with MOG(34-56). Monkeys sensitized against the viral peptide and subsequently challenged with MOG(34-56) display histological signs of encephalitis, but do not show overt neurological signs. PMID- 17126918 TI - Ribosomal therapy in the prophylaxis of recurrent pharyngotonsillitis in children. AB - OBJECTIVE: Although much has been written on how to manage recurrent pharyngotonsillitis, it remains a controversial topic. The composition of normal commensal oropharynx bacteria may be disrupted by frequently using antimicrobials, by inhibiting sensitive organisms and by allowing resistant organisms to overgrowth. This may cause the recurrence of acute episodes. The aim of this study was to evaluate the efficacy of ribosomal immunotherapy in the prophylaxis of recurrent pharyngotonsillitis. METHODS: A total of 160 children aged between 5 and 14 years with recurrent pharyngotonsillitis were ramdomized to receive either ribosomal immunotherapy (group A one tablet a day, 8 days a month for 3 months) or a placebo (group B same dosage for the same period). RESULTS: At the end of the study, each patient treated with Immucytal presented a subjective decrease of symptoms. Compared with group B, group A experienced a significant improvement of some clinical parameters. CONCLUSIONS: The results show that ribosomal immunotherapy causes a significant improvement of both specific and non specific immunity and may be effective in the prophylaxis of recurrent pharyngotonsillitis and in preventing recurrences without entailing side effects or bacterial resistance. PMID- 17126919 TI - Detection of Colorado Tick Fever viral RNA in acute human serum samples by a quantitative real-time RT-PCR assay. AB - A quantitative real-time RT-PCR assay for the detection of Colorado Tick Fever (CTF) viral RNA in human clinical samples is presented. The sensitivity of this assay has been shown to be greater than that of the isolation of virus in Vero cells by standard plaque assay in a direct comparison. The specificity of the CTF quantitative real-time RT-PCR assay was determined by the exclusive detection of CTF viral RNAs when applied to a diverse panel of CTF viral isolates and reference strain agents known to circulate in areas of CTF virus transmission. Lastly, the quantitative real-time RT-PCR assay demonstrated exceptional sensitivity for the detection of CTF viral RNA in acute human serum. The quantitative real-time RT-PCR assay is efficient, sensitive and specific and as such is useful for the detection of CTF viral RNA in the diagnostic or research laboratory. PMID- 17126920 TI - Interplay between protein synthesis and degradation in the CNS: physiological and pathological implications. AB - Compromise of the ubiquitin-proteasome system (UPS) is a potential basis for multiple physiological abnormalities and pathologies in the CNS. This could be because reduced protein turnover leads to bulk intracellular protein accumulation. However, conditions associated with compromised UPS function are also associated with impairments in protein synthesis, and impairment of UPS function is sufficient to inhibit protein synthesis. These data suggest that the toxicity of UPS inhibition need not depend on gross intracellular protein accumulation, and indicate the potential for crosstalk between the UPS and protein-synthesis pathways. In this review, we discuss evidence for interplay between the UPS and protein-synthesis machinery, and outline the implications of this crosstalk for physiological and pathological processes in the CNS. PMID- 17126921 TI - 5-HT in the enteric nervous system: gut function and neuropharmacology. AB - In recent times, the perception of functional gastrointestinal disorders such as irritable bowel syndrome (IBS) has shifted fundamentally. Such disorders are now thought of as serious diseases characterized by perturbations in the neuronal regulation of gastrointestinal function. The concept of visceral hypersensitivity, the characterization of neuronal networks in the 'brain-gut axis' and the identification of several novel 5-HT-mediated mechanisms have contributed to this shift. Here, we review how some of the more promising of these new mechanisms (e.g. those involving 5-HT transporters and the 5-HT(2B), 5 HT(7) and putative 5-HT(1p) receptors) might lead to a range of second-generation therapies that could revolutionize the treatment of functional gastrointestinal disorders, particularly IBS. PMID- 17126922 TI - Investigating the hedonic effects of interferon-alpha on female rats using brain stimulation reward. AB - Interferon-alpha (IFN-alpha) is used as a front-line treatment for cancer and other diseases. Reports of depression as a consequence of IFN-alpha therapy scatter the literature, generating interest in the CNS disruptions elicited by this cytokine. In the present work, we investigated the short- and long-term effects of a single systemic injection of vehicle, 10, or 1000 units of IFN-alpha on temperature, body weight, food intake, sickness behaviours, locomotor activity, and brain stimulation reward (BSR) thresholds elicited from the ventral tegmental area in female Long-Evans rats. Pioneered for studying motivational processes, BSR has been exploited as a tool for tracking hedonic status in animal models of depression. In this study, the main findings were that IFN-alpha did not induce anhedonia as defined by no increase in frequency thresholds. However, the analyses of sickness behaviours unveiled a significant increase in piloerection in all sham control animals that received an IFN-alpha injection while the BSR animal scores remained relatively unchanged between pre- and post injection days. This pattern was also evident in the overall total sickness behaviour scores. Our data suggest that a single exposure to IFN-alpha treatment in female rats elicits long-term somatic effects, without altering hedonic status. PMID- 17126923 TI - Immunomodulation of Mytilus hemocytes by individual estrogenic chemicals and environmentally relevant mixtures of estrogens: in vitro and in vivo studies. AB - Endocrine disrupting compounds (EDCs) are almost ubiquitous in the aquatic environment. In the marine bivalve Mytilus the natural estrogen 17beta-estradiol (E2) and different EDCs have been recently demonstrated to affect the function of the immune cells, the hemocytes. The effects were Tamoxifen-sensitive and were mediated by rapid modulation of kinase-mediated transduction pathways. In this work we compared the in vitro effects of individual estrogenic chemicals (E2, EE: 17alpha-ethynyl estradiol; MES: mestranol; NP: nonylphenol; NP1EC: nonylphenol monoethoxylate carboxylate; BPA: bisphenol A; BP: benzophenone) on hemocyte parameters: lysosomal membrane stability (LMS), phagocytosis, lysozyme release. LMS was the most sensitive effect parameter, showing a decreasing trend at increasing concentrations of estrogens. EC50 values obtained from LMS data were utilized to calculate the estradiol equivalency factor (EEF) for each compound; these EEFs allowed for an estimation of the estrogenic potential of a synthetic mixture with a composition very similar to that previously found in waters of the Venice lagoon. Concentrated mixtures significantly affected hemocyte parameters in vitro and the effects were prevented by Tamoxifen. Significant effects of the mixture were also observed in vivo, at longer exposure times and at concentrations comparable with environmental exposure levels. The results indicate that Mytilus immune parameters can be suitably utilized to evaluate the estrogenic potential of environmental samples. PMID- 17126924 TI - Growth hormone induces apelin mRNA expression and secretion in mouse 3T3-L1 adipocytes. AB - Recently, apelin was characterised as a novel adipose-expressed factor which is upregulated in rodent and human obesity and influences cardiovascular function, as well as insulin secretion. To clarify expression and regulation of this adipokine, apelin mRNA was measured by quantitative real-time reverse transcription-polymerase chain reaction in mouse 3T3-L1 adipocytes after treatment with various hormones known to induce insulin resistance. Interestingly, apelin synthesis was significantly upregulated by growth hormone (GH) and insulin in these cells whereas TNFalpha and isoproterenol did not have any effect. Thus, 500 ng/ml GH acutely induced apelin mRNA by up to 4-fold in a time-dependent fashion with significant stimulation seen at concentrations as low as 5 ng/ml effector. Furthermore, apelin secretion was assessed by enzyme-linked immunoassay in mouse adipocytes. Here, secretion of this adipokine was induced 2.85-fold by GH. Studies using pharmacological inhibitors suggested that the positive effect of GH on apelin mRNA synthesis is at least in part mediated by janus kinase 2 and phosphatidylinositol 3-kinase. Taken together, our results show a significant induction of apelin mRNA synthesis and protein secretion by GH. PMID- 17126925 TI - Relaxation induced by N-terminal fragments of chromogranin A in mouse gastric preparations. AB - A definitive role for chromogranin A (CGA)-derived fragments in the control of the gastrointestinal smooth muscle contractility has not been yet established. The purpose of the present study was to evaluate, in vitro, the effects of the recombinant vasostatin 1-78 (VS-1), CGA 7-57 and CGA 47-66 on the mouse gastric mechanical activity, recording the changes of intraluminal pressure. VS-1, CGA 7 57 and CGA 47-66 produced concentration-dependent relaxations. Mouse anti vasostatin-1 monoclonal antibody 5A8, recognising the region 53-57, abolished the relaxation induced by VS-1, indicating the specificity of the effect. The relaxation was significantly reduced by tetrodotoxin (TTX), blocker of neuronal voltage-dependent Na(+) channels, l-NAME, inhibitor of nitric oxide (NO) synthase, or apamin, blocker of small conductance Ca(2+)-dependent K(+) channels. The joint application of TTX and l-NAME did not show any additive effects, whereas TTX plus apamin abolished the VS-1 response. The results suggest that the N-terminal CGA-derived peptides are able to relax mouse gastric muscle and, therefore, they point out an inhibitory role of vasostatin I in the gastrointestinal tract. The relaxation is mediated in part by neural mechanisms through NO production and in part by non-neural mechanisms involving the opening of small conductance Ca(2+)-dependent K(+) channels. PMID- 17126926 TI - Closure of a home hospital program: impact on hospitalization rates. AB - Home hospitalization (HH), as a substitute to in-patient care, is an area of growing interest, particularly amongst the elderly. Debate nonetheless exists concerning its economic justification. This study describes a natural experiment that arose following spending cuts and closure of the 400 patient Jerusalem HH program. It examines the hypothesis that HH closure would cause increasing geriatric and general medical hospital utilization amongst the 45,000 beneficiaries of the Jerusalem Clalit Health Fund (HMO) aged 65 years and over. Hospitalization rates were measured prior to and following HH closure, and analysis of variance confirmed the significance of the differences in both geriatric (p<0.0001) and general medical hospitalization rates (p=0.02) over the study period. Linear regression analyses of the hospitalization rates prior to HH closure were performed to determine the expected trajectory of hospitalization rates following HH closure. The observed hospital utilization in the year following HH closure cost 6.2 million US dollars in excess of predicted expenditure; closure of the HH resulted in the saving of 1.3 million USdollars. The ratio of direct increased costs to savings was 5:1 thus confirming the hypothesis that HH closure would result in increased hospital utilization rates among the local elderly population. PMID- 17126927 TI - Kawasaki disease resembling a retropharyngeal abscess--case report and literature review. AB - A retropharyngeal abscess is a rare presentation of Kawasaki disease. Herein, we present a case of Kawasaki disease mimicking a retropharyngeal abscess, and review five previously published case reports. Once the correct diagnosis of Kawasaki disease is established, any abscess-like lesion on computed tomography may be just a false-positive finding. PMID- 17126928 TI - Serum vascular endothelial growth factor in cyanotic congenital heart disease functionally contributes to endothelial cell kinetics in vitro. AB - BACKGROUND: Remarkable amounts of neovascularization develop in patients with cyanotic congenital heart disease who have low pulmonary blood flow and systemic cyanosis, but the factors functionally responsible for angiogenesis in cyanotic congenital heart disease have not been determined. METHODS AND RESULTS: To investigate the functional angiogenic molecules in serum from these patients, serum angiogenic activity was studied in 21 patients (systemic oxygen saturation: 82+/-1.9%) and in 17 healthy controls. Patient serum was more active in stimulating the tube formation of human umbilical vein endothelial cells (HUVECs) into capillary-like structures than control serum (150% vs 104% of internal control; p<0.001). This increased serum angiogenic activity normalized after total cardiac repair (p<0.001). The migration activity of HUVECs was also accelerated in patient serum (p=0.007). To identify the molecules in patient serum affecting tube formation of HUVECs, we examined the effects of an inhibitor or a neutralizing antibody against various angiogenic molecules on in vitro angiogenesis. Both the soluble vascular endothelial growth factor (VEGF) receptor 1 and the VEGF receptor 2 tyrosine kinase inhibitor SU5416 reduced the basal serum angiogenic activity of patients and this was reversed by a supplement of recombinant human VEGF. CONCLUSION: Our results indicate that serum VEGF functionally contributes to vascular endothelial cell kinetics in patients with cyanotic congenital heart disease. PMID- 17126929 TI - Previously undiagnosed congenitally corrected transposition of the great arteries in a 51-year-old woman with chronic heart failure symptoms. AB - We describe the case of a 51-year-old female with previously undiagnosed congenitally corrected transposition of the great arteries (ccTGA) who was admitted to the hospital due to the exacerbated symptoms of heart failure for planned coronary angiography. Patient cardiac functional status appeared to have gradually regressed to New York Heart Association class III at the time of the admission. The chart reviews were performed. Coronary angiography did not reveal the occlusion of the coronary artery. Radiography, echocardiography, and multislice computed tomography confirmed newly diagnosed ccTGA. PMID- 17126930 TI - Lone atrial fibrillation and sport practice. The no gain without pain history revisited again? PMID- 17126931 TI - A complete metal jacket case using ten paclitaxel-eluting stents for multiple de novo coronary artery lesions. AB - Although full metal jacket using drug-eluting stent (DES) for a single coronary artery disease has sparsely been described before, there is no report of safety and efficacy of complete metal jacket from left main (LM) to three major coronary arteries. We report a complete metal jacket case using 10 paclitaxel-eluting stents (PES; Taxus, Boston Scientific) for a triple vessel diffuse de novo coronary diseases who refused coronary artery bypass graft. PMID- 17126932 TI - 12 lead ECG in the detection of left main coronary artery occlusion during acute coronary syndromes. AB - A careful evaluation of the 12 Lead surface ECG may help detect Left Main Coronary artery (LMCA) occlusion. Since LMCA occlusion can cause rapid hemodynamic and electrical deterioration, early identification may help the treating team to plan out timely revascularization. We describe a 61 years old male who presented with acute anterior wall myocardial infarction and where LMCA occlusion was suspected on the basis of surface ECG. The diagnostic criteria available are discussed. PMID- 17126934 TI - A calcified cardiac mass. AB - Cardiac fibromas are benign tumours, often diagnosed in childhood, but rarely they may be diagnosed in adults or the elderly. We present an interesting case of a middle-aged lady presenting with exertional chest pain and breathlessness, who was found to have a heavily calcified mass within the myocardium. With a previous history of chest trauma, a calcified myocardial haematoma was initially suspected. Complete surgical excision led to a total resolution of symptoms. Histological examination confirmed the diagnosis of a cardiac fibroma. Complete excision of cardiac fibromas, where possible, is advised and is associated with excellent survival. PMID- 17126935 TI - Percutaneous coronary intervention through a Cabrol composite graft. AB - There are several different surgical techniques used to repair acute type-A aortic dissections. One such technique is called the Cabrol procedure which uses a Dacron tube graft to re-attach the native coronary arteries. Stenosis of this graft-native coronary anastomosis is a recognized complication of the procedure and one amenable to percutaneous intervention. However, knowledge of the complex anatomy involved with the Cabrol repair is required in order to potentially carry out successful intervention. We report one such case involving a critical stenosis at the anastomosis of the Cabrol graft with the native left main stem. PMID- 17126936 TI - Assessment of QTc interval dispersion in patients with chronic heart failure secondary to Chagas-Hypertensive cardiomyopathy. AB - No previous study has examined the association of clinical variables and QTc interval dispersion in patients with Chagas-Hypertensive cardiomyopathy. Sixty four patients with Chagas-Hypertensive cardiomyopathy were entered for the study. Seventy-one chronic Chagas' disease heart failure patients and 68 patients with Hypertensive cardiomyopathy heart failure were used in the comparison of QTc interval dispersion. The QTc interval dispersion was defined as the difference between maximum and minimum QTc. Median QTc interval dispersion was 75 (55) ms in patients with Chagas' cardiomyopathy, (56) 73 ms in patients with Hypertensive cardiomyopathy, and (87.3) 98 ms in patients with Chagas-Hypertensive cardiomyopathy (p>0.05). In patients with Chagas-Hypertensive cardiomyopathy, median QTc interval was (134) 100 ms in survivors and (87) 96 ms in nonsurvivors (p>0.05). Thus, QTc interval dispersion values in patients with Chagas Hypertensive cardiomyopathy are not associated with mortality and similar to those found in patients with Chagas' cardiomyopathy and in patients with Hypertensive cardiomyopathy. PMID- 17126937 TI - Quantification of functional genes from procaryotes in soil by PCR. AB - Controlling turnover processes and fluxes in soils and other environments requires information about the gene pool and possibilities for its in situ induction. Therefore in the recent years there has been a growing interest in genes and transcripts coding for metabolic enzymes. Besides questions addressing redundancy and diversity, more and more attention is given on the abundance of specific DNA and mRNA in the different habitats. This review will describe several PCR techniques that are suitable for quantification of functional genes and transcripts such as MPN-PCR, competitive PCR and real-time PCR. The advantages and disadvantages of the mentioned methods are discussed. In addition, the problems of quantitative extraction of nucleic acid and substances that inhibit polymerase are described. Finally, some examples from recent papers are given to demonstrate the applicability and usefulness of the different approaches. PMID- 17126938 TI - Uncertainties when using only one MRI-based treatment plan for subsequent high dose-rate tandem and ring applications in brachytherapy of cervix cancer. AB - BACKGROUND AND PURPOSE: This retrospective study compares individual MRI based 3D treatment planning for each intracavitary applicator insertion and the use of only one MRI treatment plan for cervical cancer brachytherapy. MATERIALS AND METHODS: GTV, high risk (HR) CTV and OAR were delineated and analysed for 14 patients. Data using the individual approach were taken from the actual irradiated plans. The "single plan procedure" was simulated by matching the dose distribution of the first plan to the MRI datasets of each subsequent implantation. Total doses from brachytherapy were added up and normalized to 2Gy fractionation (EQD2). RESULTS: The mean D90 for HR CTV was 6Gy higher when using one plan than when using individual treatment plans. The D(2cc) increased 3.5Gy for the bladder, 4.2Gy for the rectum and 5.8Gy for the sigmoid. The use of only one treatment plan would have resulted in 2, 1 and 5 extra cases exceeding total D(2cc) constraints for bladder (90Gy), rectum (75Gy) and sigmoid (75Gy), respectively. CONCLUSION: The use of only one treatment plan for several applications results in higher dose to target and OAR structures. CT, clinical examination and X-ray findings can help to reduce certain situations of overdosage, when individual MRI based treatment planning is not available for each fraction. PMID- 17126939 TI - Hormone receptor status as an independent variable affecting the survival of advanced breast cancer patients with brain metastases. PMID- 17126940 TI - Accelerated partial breast irradiation with multi-catheter brachytherapy: Local control, side effects and cosmetic outcome for 274 patients. Results of the German-Austrian multi-centre trial. AB - BACKGROUND AND PURPOSE: To evaluate the safety and effectiveness of accelerated partial breast irradiation (APBI) with multi-catheter brachytherapy. PATIENTS AND METHODS: Between 11/2000 and 04/2005, 274 patients participated. Patients were eligible for APBI if they had histologically confirmed breast cancer, a tumour diameter 3cm, complete resection with clear margins 2mm, negative axillary lymph nodes or singular nodal micro-metastasis, no distant metastases, hormone receptor positive tumours, and were >/= 35 years. Patients were excluded if mammographically they showed a multicentric invasive growth pattern, poorly differentiated tumours, residual diffuse micro-calcifications, extensive intraductal component, or vessel invasion. Median follow-up was 32 months (range; 8-68). RESULTS: The local control rate was 99.3% (272/274), the 3 year-local recurrence-free survival probability, 99.6%. Peri-operative complications occurred in 5.5% (15/274): 9 (3.3%) experienced implant infection and 6 (2.2%) hematoma. Acute toxicity (Grade 1/2 radio-dermatitis) was seen in 6.6% (18/274). Late side effects >/= Grade 3 (fibrosis, telangiectasia) occurred in 1.8% (4/274). Cosmetic results were excellent/good in 94% (253/274). CONCLUSIONS: This analysis underlined the safety and effectiveness of APBI in a carefully selected subgroup with favourable disease characteristics. Of course, longer follow-up and randomised trials are necessary to conclusively assess the potential of APBI. PMID- 17126941 TI - Kinematic analysis of multiple constraints on a pointing task. AB - The current experiment suggests that the speed/accuracy tradeoff is composed of two classes of constraints, effector and task. We examined the effects of movement distance, target size, orientation of the movement in the workspace, and C-D gain on the kinematics of discrete pointing movements made with computer mouse. It was found that target size influenced the shape of velocity profiles by elongating the duration of the corrective sub-movement phase, while movement distance scaled the entire velocity curve without affecting its shape. C-D gain and orientation of the movement exhibited two kinds of effects: an overall scaling of the velocity curve and a change in its shape. We conclude that target size is a task constraint and movement distance is an effector constraint, while movement orientation exhibited characteristics of both. C-D gain by itself was not a constraint, but interacted with both task and effector constrains. These results highlight the roles of biomechanical and information processing factors in the speed/accuracy tradeoff. PMID- 17126942 TI - Swimming constraints and arm coordination. AB - Following Newell's concept of constraint (1986), we sought to identify the constraints (organismic, environmental and task) on front crawl performance, focusing on arm coordination adaptations over increasing race paces. Forty-two swimmers (15 elite men, 15 mid-level men and 12 elite women) performed seven self paced swim trials (race paces: as if competitively swimming 1500m, 800m, 400m, 200m, 100m, 50m, and maximal velocity, respectively) using the front crawl stroke. The paces were race simulations over 25m to avoid fatigue effects. Swim velocity, stroke rate, stroke length, and various arm stroke phases were calculated from video analysis. Arm coordination was quantified in terms of an index of coordination (IdC) based on the lag time between the propulsive phases of each arm. This measure quantified three possible coordination modes in the front crawl: opposition (continuity between the two arm propulsions), catch-up (a time gap between the two arm propulsions) and superposition (an overlap of the two arm propulsions). With increasing race paces, swim velocity, stroke rate, and stroke length, the three groups showed a similar transition in arm coordination mode at the critical 200m pace, which separated the long- and mid-pace pattern from the sprint pace pattern. The 200m pace was also characterized by a stroke rate close to 40strokemin(-1). The finding that all three groups showed a similar adaptation of arm coordination suggested that race paces, swim velocity, stroke rate and stroke length reflect task constraints that can be manipulated as control parameters, with race paces (R(2)=.28) and stroke rate (R(2)=.36) being the best predictors of IdC changes. On the other hand, only the elite men reached a velocity greater than 1.8ms(-1) and a stroke rate of 50strokemin(-1). They did so using superposition of the propulsion phases of the two arms, which occurred because of the great forward resistance created when these swimmers achieved high velocity, i.e., an environmental constraint. Conversely, the elite women and mid level men had shorter stroke lengths and maintained a time gap between the propulsions of the two arms throughout the increase in paces, with gender and expertise explaining 9% and 8.3% of the IdC changes, respectively. These results indicate that arm coordination cannot be interpreted solely from the IdC value but should be considered from the perspective of task, environmental, and organismic constraints. These constraints can serve as control parameters in experiments aimed at gaining insight into changes in arm coordination during the front crawl. In this context, catch-up coordination, which is often considered as a mistake, was seen to be an adaptation to a relative constraint. PMID- 17126943 TI - Fermentation process for tetrameric human collagen prolyl 4-hydroxylase in Escherichia coli: improvement by gene optimisation of the PDI/beta subunit and repeated addition of the inducer anhydrotetracycline. AB - The collagen prolyl 4-hydroxylases (C-P4Hs) that reside within the lumen of the endoplasmic reticulum (ER) are the key enzymes in the biosynthesis of collagens. The vertebrate enzymes are alpha(2)beta(2) tetramers consisting of two catalytic alpha subunits and two beta subunits that are identical to protein disulfide isomerase (PDI). Cytoplasmic production of an active human C-P4H has recently been described in the Origami (trxB gor) mutant Escherichia coli using a bicistronic vector with independent control of the alpha and PDI/beta subunit expression by the tetA and T5-lac promoters, respectively, enabling sequential induction (Neubauer, A., Neubauer, P., Myllyharju, J., 2005. High-level production of human collagen prolyl 4-hydroxylase in Escherichia coli. Matrix Biol. 24, 59-68). We show here that the yield of active C-P4H in shake flasks is increased 50-fold by improving the expression level of the PDI/beta subunit through gene optimisation. We also found that stable expression of the alpha subunit mRNA in a fed-batch fermentation process requires repeated additions of anhydrotetracycline. This finding may be of a wider general importance for the use of the tetA promoter in fed-batch cultivations, especially if recombinant proteins are expressed during long production phases. We also show that growth of the E. coli Origami strain to high cell density on a complex medium with consecutive sequential induction is difficult to achieve and that optimisation of similarly complicated systems can greatly benefit from the use of quantitative mRNA analysis for the evaluation of transcriptional bottlenecks. The optimisation approach resulted in a fermentation yield of 143 mg L(-1) of active C-P4H, corresponding to approximately 7.5% of the total soluble cell protein. PMID- 17126944 TI - High avidity binding of engineered papaya mosaic virus virus-like particles to resting spores of Plasmodiophora brassicae. AB - Papaya mosaic virus (PapMV) like particles (VLPs) were used as a platform for fusion of affinity peptides binding to resting spores of Plasmodiophora brassicae a major pathogen of crucifers. Three peptides with specific affinity to the target were isolated and cloned at the C-terminus of the PapMV coat protein (CP), generating three different high avidity VLPs. The peptides were exposed at the surface of the VLPs and their avidity to resting spores of P. brassicae was measured by flow cytometry. NLP-A, with the peptide DPAPRPR, showed the highest avidity. The binding avidity of NLP-A to P. brassicae spores was comparable to that of a polyclonal antibody. NLP-A was also shown to be more specific than the antibody. Fusion of the affinity peptide to a monomeric form (mCP) of the CP [Lecours, K., Tremblay, M.-H., Laliberte Gagne, M.-E., Gagne, S.M., Leclerc, D., 2006. Purification and biochemical characterization of a monomeric form of papaya mosaic potexvirus coat protein. Protein Express. Purific. 47, 273-280] generated a fusion protein that was unable to assemble into VLPs, and mCP-A fusions failed to bind resting spores. The avidity of VLP-A was increased by adding a glycine spacer between the C-terminus of the PapMV CP and the peptide, and improved even further by using a duplicated A peptide in the fusion protein. The use of high avidity VLPs has advantages over polyclonal antibodies because of target specificity. VLPs offers the specificity of monoclonal antibodies but can be more easily generated using the powerful selection of phage display. PMID- 17126945 TI - Sustained release of bioactive therapeutic proteins from a biodegradable elastomeric device. AB - Effective localized delivery of a therapeutic protein requires a biodegradable device capable of delivering active protein at a sustained rate, and at a concentration within its therapeutic window. The objective of this study was to demonstrate that a biodegradable elastomeric device can be made in a cylindrical geometry, and still retain the ability to release a variety of therapeutic proteins at a nearly constant rate in nanomolar concentration with high bioactivity. The elastomers were prepared with cylindrical geometry by photo cross-linking an acrylated star-poly(epsilon-caprolactone-co-d,l-lactide) macromer. Vascular endothelial growth factor (VEGF), interferon-gamma (IFN gamma), and interleukin-2 (IL-2) were co-lyophilized with excipients, then entrapped within the elastomer matrix by photo-polymerization. Under identical formulation conditions, these proteins were released at the same, nearly constant rate for a significant part of the release profile (until 70%-80% release depending on formulation characteristics). Decreasing the molecular weight of the acrylated macromer increased the rate of protein release, but did not alter the zero order nature of the release kinetics. Cell based bioactivity assays showed only that 57% of the VEGF released was bioactive. By contrast, both IL-2 and IFN gamma showed relatively high bioactivity and over 80% of the released proteins were bioactive. The elastomer formulation has potential as a regio-specific protein delivery device. PMID- 17126946 TI - Antepartum and intra-partum insulin management of type 1 and type 2 diabetic women: Impact on clinically significant neonatal hypoglycemia. AB - OBJECTIVE: Assess efficacy of intrapartum intravenous (i.v.) insulin and effect of antepartum and intrapartum diabetic control on various measures of clinical neonatal hypoglycemia. DESIGN: Retrospective chart review. SETTING: Regional Diabetes in Pregnancy Clinic. RESULTS: Maternal hypoglycemia occurred intrapartum in 56% of subjects. Mean delivery blood glucose with intravenous insulin use was 6.7mmol/l. Capillary blood glucose <2.2mmol/l occurred in 69% of neonates and 44% received intravenous glucose. Maternal delivery glucose >6.5mmol/l correlated with occurrence of neonatal glucose <2.2mmol/l but not requirement for i.v. glucose or NICU admission. A third trimester HbA1c >6.5% had a stronger association with NICU admission and i.v. glucose requirement. CONCLUSIONS: Blood glucose at delivery >6.5mmol/l predicts neonatal hypoglycemia but does not correlate with severity. Chronic maternal hyperglycemia, reflected by pre delivery HbA1c, predicts severe fetal hyperinsulinism and requirement for aggressive intervention. This stresses the importance of a target third trimester HbA1c of <6.5%. PMID- 17126947 TI - Population impact--definition, calculation and its use in prevention science in the example of tobacco smoking reduction. AB - INTRODUCTION: Population Impact is a criterion that can enhance prevention practices and provide a solid foundation for integrating policies and programs for prevention. However, to quantify the population impact of programs a statistical measure is needed. The objective of this article is to (a) deduct a formula to quantify population impact (PI), (b) define the formula for population impact of smoking prevention measures and (c) apply this formula on smoking prevention programs. METHODS: Decision analytical approach. RESULTS: The measurement of PI is defined with four parameters: recruitment, retention, efficacy and prevalence. A formula is mathematically deducted and the PI for different smoking prevention programs is calculated. DISCUSSION: The formula supports decision makers in deciding what prevention measure shows a higher impact on the population, gives hints where to improve the measure to increase the impact, whether recruitment, retention or efficacy needs to be improved and makes it easy to do analyses of costs on the population level. CONCLUSIONS: To enhance prevention practice prevention measures need to provide all parameters to calculate the PI, research needs to focus on all parameters influencing the PI and costs of prevention measures need to be provided. PMID- 17126948 TI - Common evolutionary trends for SINE RNA structures. AB - Short interspersed elements (SINEs) and long interspersed elements (LINEs) are transposable elements in eukaryotic genomes that mobilize through an RNA intermediate. Understanding their evolution is important because of their impact on the host genome. Most eukaryotic SINEs are ancestrally related to tRNA genes, although the typical tRNA cloverleaf structure is not apparent for most SINE consensus RNAs. Using a cladistic method where RNA structural components were coded as polarized and ordered multistate characters, we showed that related structural motifs are present in most SINE RNAs from mammals, fishes and plants, suggesting common selective constraints imposed at the SINE RNA structural level. Based on these results, we propose a general multistep model for the evolution of tRNA-related SINEs in eukaryotes. PMID- 17126949 TI - Another role for RNA: a messenger across generations. AB - There has been a recent resurgence of interest in the notion that DNA is not the sole determinant of our inherited phenotype. The strongest evidence for transgenerational epigenetic inheritance has come from studies of paramutation in plants. But few examples have been reported in other species, and the molecular basis for the process has been unclear. A recent paper by Rassoulzadegan and colleagues provides clear evidence of paramutation in mice and strong evidence that RNA is involved. PMID- 17126950 TI - MUC4 expression correlates with poor prognosis in small-sized lung adenocarcinoma. AB - The mortality of lung cancer remains high, despite improved diagnostic techniques that allow small lung tumors to be detected. In this study, we evaluated the prognostic significance of the tracheal mucin MUC4 by immunohistochemical investigation of the expression profiles of MUC4, ErbB2, p27 and MUC1 in lung adenocarcinoma specimens (non-bronchiolo-alveolar type, < or =3cm) from 185 patients. MUC4 is a membrane mucin, similarly to MUC1, and in addition MUC4 functions as an intra-membrane ligand for receptor tyrosine kinase ErbB2 and is associated with regulation of p27. However, MUC4 expression was found to be unrelated to expression of MUC1, ErbB2 and p27 in small-sized lung adenocarcinomas. The disease-free interval (DFI) and survival rate of 25 patients with high MUC4 expression (> or =25% of neoplastic cells stained) were significantly lower than those of 160 patients with low MUC4 expression (<25% of neoplastic cells stained) (P<0.05), whereas ErbB2 and p27 expression showed no significant correlation with DFI and survival. Univariate analysis showed that high MUC4 and p27 expression correlated with blood vessel invasion (P=0.0004), and MUC4 expression was frequently detected in regions of stromal invasion. In addition, the survival rate of stage IA patients with high MUC4 expression was significantly lower than that of stage IA patients with low MUC4 expression (P<0.05). In conclusion, high MUC4 expression in small-sized lung adenocarcinomas correlates with a short DFI and a poor survival rate. Therefore, MUC4 expression might be a new independent factor for prediction of outcome and indication of poor prognosis in lung adenocarcinoma. PMID- 17126951 TI - Randomized trial of drip infusion versus bolus injection of vinorelbine for the control of local venous toxicity. AB - Vinorelbine is a moderate vesicant that is well known to cause local venous toxicity such as drug induced-phlebitis. We conducted a prospective randomized trial to determine whether a 1-min bolus injection (1 min bolus) of vinorelbine reduced the incidence of local venous toxicity compared with a 6-min drip infusion (6 min infusion). Non-small cell lung cancer patients who were to receive chemotherapy containing vinorelbine were randomly assigned to receive either 6 min infusion or 1 min bolus of the drug. All infusions were administered through a peripheral vein. Local venous toxicity was evaluated at each infusion up to two cycles. Eighty-three patients were randomized into the study and 81 of them assessable for analysis. One hundred thirty-eight infusions to 40 patients in 6 min infusion and 135 infusions to 41 patients in 1 min bolus were delivered. Vinorelbine induced-local venous toxicity was observed in 33% of patients in 6 min infusion and 24% in 1 min bolus. There was no statistically significant difference between the two arms (P=0.41). The incidence of local venous toxicity per infusions was 16% (22 of 138 infusions) in 6 min infusion and 11% (15 of 135 infusions) in 1 min bolus (P=0.47). No severe local venous toxicity was seen in either arm. In this study, the administration of in 1 min bolus of vinorelbine did not significantly reduce the incidence of local venous toxicity compared with 6 min infusion. Further studies for the control of local venous toxicity of vinorelbine are warranted. PMID- 17126952 TI - Gastrin 1-6 promotes growth of colon cancer cells through non-CCK receptors. AB - Our previous studies have shown that stimulation of proliferation of DLD-1 and HT29 human colonic cancer cells by nanomolar gastrin (G17) and carboxymethyl gastrin (G17Gly) and reversal of growth by micromolar G17 and G17Gly involves binding sites which can neither be CCK1 nor CCK2 receptors; the N terminal fragment, G17(1-12), is sufficient to increase the number of HT-29 cells by binding the higher affinity binding site but is without a suppressing effect through the lower affinity site. In this study with DLD-1 cells, competitive binding using 125I-G17(1-12) showed that G17(1-12) binds both high and low affinity sites, as do G17 and G17Gly. G17(1-6)-NH2, even without the central-to-C terminal portion of G17, was still able to bind a single site and to promote a dose-dependent increase in cell number at nanomolar concentrations. The results indicate the presence of a non-CCK receptor on human colonic cancer cells which could mediate the tumor-promoting activity of the N-terminal-to-central portion of G17Gly which, unlike G17, is produced by such cells. PMID- 17126953 TI - Phospholipids modulate superoxide and nitric oxide production by lipopolysaccharide and phorbol 12-myristate-13-acetate-activated microglia. AB - Microglial activation and inflammatory processes have been implicated in the pathogenesis of a number of neurodegenerative disorders. Recently, peroxynitrite (ONOO(-)), the reaction product of superoxide (O(2)(-)) and nitric oxide (NO) both of which can be generated by activated microglia, has been demonstrated to act as a major mediator in the neurotoxicity induced by activated microglia. On the other hand, phospholipids such as phosphatidylserine (PS) and phosphatidylcholine (PC) have been reported to modulate the immune function of phagocytes. We therefore evaluated the effects of liposomes which comprise both PS and PC (PS/PC liposomes) or PC only (PC liposomes) regarding the production of both O(2)(-) and NO by lipopolysaccharide (LPS)/phorbol 12-myristate-13-acetate (PMA)-activated microglia using electron spin resonance (ESR) spin trap technique with a DEPMPO and Griess reaction, respectively. Pretreatment with PS/PC liposomes or PC liposomes considerably inhibited the signal intensity of O(2)(-) adduct associated with LPS/PMA-activated microglia in a dose-dependent manner. In addition, pretreatment with PS/PC liposomes also significantly reduced LPS/PMA induced microglial NO production. In contrast, pretreatment with PC liposomes had no effect on the NO production. These results indicate that PS/PC liposomes can inhibit the microglial production of both NO and O(2)(-), and thus presumably prevent a subsequent formation of ONOO(-). Therefore, PS/PC liposomes appear to have both neuroprotective and anti-oxidative properties through the inhibition of microglial activation. PMID- 17126954 TI - Altered navigational strategy use and visuospatial deficits in hAPP transgenic mice. AB - Navigation deficits are prominent in Alzheimer's disease (AD) patients and transgenic mice expressing familial AD-mutant hAPP and A beta peptides. To determine the impact of strategy use on these deficits, we assessed hAPP and nontransgenic mice in a cross maze that can be solved by allocentric (world based) or egocentric (self-based) strategies. Most nontransgenic mice used allocentric strategies, whereas half of hAPP mice were egocentric. At 3 months, all mice learned the cross maze rapidly; at 6 months, only allocentric hAPP mice were impaired. At 3 and 6 months, hAPP mice had reduced hippocampal Fos expression, which correlated with cross maze learning in older mice. Striatal pCREB expression was unaltered in hAPP mice, suggesting striatal sparing. We conclude that egocentric strategy use may be an earlier indicator of hAPP/A beta induced hippocampal impairment than spatial learning deficits. Persistent use of allocentric strategies when egocentric strategies are available is maladaptive when there is hippocampal damage. Interventions promoting flexibility in selecting learning strategies might help circumvent otherwise debilitating navigational deficits caused by AD-related hippocampal dysfunction. PMID- 17126955 TI - Exploring haematology nurses' perceptions of specialist education's contribution to care delivery and the development of expertise. AB - The role that educational preparation may play in the delivery of care and the development of expertise is a point of some debate [Manley, K., Garbett, R., 2000. Paying Peter and Paul reconciling concepts of expertise with competency for a clinical career structure. Journal of Clinical Nursing 9 (3), 347; King, L., Macleod Clark, J., 2002. Intuition and the development of expertise in surgical ward and intensive care nurses. Journal of Advanced Nursing 37(4), 322-329; Bonner, A., 2003. Recognition of expertise: an important concept in the acquisition of nephrology nursing expertise. Nursing & Health Sciences Journal 5, 123-131; Dunphy, B.C., Williamson, S.L., 2004. In pursuit of expertise. Advances in Health Sciences Education 9, 107-127]. Though education is a concept that may be universally valued, it may be more difficult to clearly discern the significance it has for practitioners who are developing their expertise. This research project employed an interpretive phenomenological design to explore the perceptions of specialist haematology nursing staff on the extent to which specialist education contributes to care delivery and the development of expert practice. A non-representative purposive sample of qualified nurses who had undertaken specialist education in haemopoiesis and work in specialist haematology participated in a focus group and semi-structured interviews. The report concludes that, for these specialist practitioners, specialist educational input had a beneficial impact on their levels of knowledge and confidence. Further to this, involvement in higher education had enabled them to become more active in the learning process. Perhaps the key finding of the study was the assertion by respondents that specialist educational input had enabled them to develop their specialist practice to a level that experience alone could not achieve. PMID- 17126956 TI - New roles to support practice learning - can they facilitate expansion of placement capacity? AB - The National Health Service (NHS) Plan [Department of Health, 2000. The NHS Plan: A Plan for Investment, a Plan for Reform, The Stationery Office, London] set out an ambitious programme of growth for the number of students undertaking health professional education programmes. To meet this demand there is a growing and widely acknowledged need to increase the number of clinical placements needed by these students. This paper outlines the findings from a regional project that aimed to map the current pattern and availability of clinical placements for healthcare students by the collection of quantitative placement data (such as location, specialty, and number of mentors) as well as in-depth interviews with Clinical Placement Managers (CPMs). This article will focus on the findings from interviews with CPMs and their views and experiences of what role they can play in increasing placement capacity. The study revealed that the introduction of this role had facilitated the development and expansion of placement capacity, as the CPMs filled a gap in the form of up-to-date local knowledge about the clinical areas. The CPMs provided a much needed 'bridge' between the Higher Education Institution (HEI), the student and the clinical area. Recommendations are made in relation to future introduction of similar roles that aim to support student learning in practice. Furthermore, useful insights for ongoing policy implementation and development are highlighted. PMID- 17126957 TI - Vaccination associated thrombocytopenic purpura in children. AB - Patients who presented with purpura and blood platelets <30x10(9)/l within 1 month after vaccination were collected from a population based material of 506 consecutive pediatric patients with newly diagnosed ITP. Of the 35 such patients, 24 had thrombocytopenia after MMR vaccination giving an estimated ITP risk of approximately 1 in 30,000 MMR inoculations. Symptoms of the 35 patients were nearly always acute. Thrombocytopenia disappeared within a month in 74% of the study patients and lasted longer than 6 months in only 10%. Bleeding episodes were uncommon during the follow-up period. We conclude that the incidence of symptomatic thrombocytopenia after vaccinations is much lower than that after respective natural infections and that the outcome in most cases is excellent. PMID- 17126958 TI - Atypical transmissible spongiform encephalopathies (TSEs) in ruminants. AB - Transmissible spongiform encephalopathies (TSEs) are associated with the accumulation in infected tissues of a disease-associated form of a host-encoded protein, the prion protein (PrP). Contrary to the normal form of the protein, this form of PrP is partially resistant to protease digestion (PrP(res)). Detailed characterisation of PrP(res) has been intensively investigated in recent years to try and decipher the diversity of TSEs in human and animals. This considerably and unexpectedly enlarged our knowledge about such diseases in ruminants. Previously, such a diversity was essentially shown by the demonstration that scrapie from sheep and goats could have different biological behaviours following transmission of the disease in mice, unlike bovine spongiform encephalopathy from cattle (BSE) which showed a distinct and unique behaviour. The properties of the BSE agent were also demonstrated to be very stable, following transmission to a variety of different species. Molecular studies of PrP(res), followed by transmission studies to mice, gave the first evidence for the accidental transmission of the BSE agent to humans where it induced a variant form of the fatal Creutzfeldt-Jakob disease (CJD) and also to different animal species including a goat in France. This last case was found among a few unusual cases of TSEs in small ruminants that showed some molecular similarities with BSE and which are currently under investigation by transmission studies in mice. The application of the molecular methods to characterise PrP(res) has most recently led to the unexpected discovery of deviant BSE forms in a few affected cattle in Europe and in the United States, which raises the question of a possible different origin at least of some cases of BSE in cattle. Finally, considerable numbers of a new TSE form in small ruminants, referred to as "atypical scrapie" or "Nor98", have meanwhile been identified in most European countries by TSE rapid testing using an assay which recognizes also comparatively less PK resistant PrP(res). PMID- 17126959 TI - Global FMD control--is it an option? AB - The outbreaks of foot-and-mouth disease (FMD) in Europe in 2001 identified the vulnerability of the intensive agricultural industries in Europe and North America to the economic consequences of the introduction of a highly infectious animal disease. The very large illegal international trade in animal products bypasses the safeguards recommended by World Animal Health Organization (OIE) and put in place by governments to prevent the importation of foreign pathogens. If it is not possible to stop the entry of FMD virus, what are the options to mitigate the risk by reducing the area of the globe in which FMD is endemic? There are a number of constraints that would prevent global control of FMD; current vaccines are expensive, have a narrow antigenic spectrum, provide only short term immunity and are very fragile; diagnostics are also expensive, require training to use and if not handled properly lose sensitivity and specificity; we still do not understand the significance of carrier animals in the epidemiology of FMD, and whether it is necessary or possible to prevent the carrier state; and many decision support tools, such as models are currently more dangerous than useful in that they fail to fully accommodate all the complexities of the disease. The four national foreign animal disease laboratories in USA, Canada, UK and Australia together with the International Livestock Research Institute have put forward a proposal to address some of these constraints (the Global FMD Research Alliance, GFRA), not only to protect their own national livestock industries, but also to support FMD control programs in countries in which the disease is present. PMID- 17126960 TI - An overview of the epidemiology of avian influenza. AB - Only viruses of the Influenzavirus A genus have been isolated from birds and termed avian influenza [AI] viruses, but viruses with all 16 haemagglutinin [H1 H16] and all 9 neuraminidase [N1-N9] influenza A subtypes in the majority of possible combinations have been isolated from avian species. Influenza A viruses infecting poultry can be divided into two groups. The very virulent viruses causing highly pathogenic avian influenza [HPAI], with flock mortality as high as 100%. These viruses have been restricted to subtypes H5 and H7, although not all H5 and H7 viruses cause HPAI. All other viruses cause a milder, primarily respiratory, disease [LPAI], unless exacerbated. Until recently HPAI viruses were rarely isolated from wild birds, but for LPAI viruses extremely high isolation rates have been recorded in surveillance studies, with overall figures of about 11% for ducks and geese and around 2% for all other species. Influenza viruses may infect all types of domestic or captive birds in all areas of the world, the frequency with which primary infections occur in any type of bird usually depending on the degree of contact there is with feral birds. Secondary spread is usually associated with human involvement, either by bird or bird product movement or by transferring infective faeces from infected to susceptible birds, but potentially wild birds could be involved. In recent years there have been costly outbreaks of HPAI in poultry in Italy, The Netherlands and Canada and in each millions of birds were slaughtered to bring the outbreaks under control. Since the 1990s AI infections due to two subtypes have been widespread in poultry across a large area of the World. LPAI H9N2 appears to have spread across the whole of Asia in that time and has become endemic in poultry in many of the affected countries. However, these outbreaks have tended to have been overshadowed by the H5N1 HPAI virus, initially isolated in China, that has now spread in poultry and/or wild birds throughout Asia and into Europe and Africa, resulting in the death or culling of hundreds of millions of poultry and posing a significant zoonosis threat. PMID- 17126961 TI - Impact of plasmid stability on oral DNA delivery by Salmonella enterica serovar Typhimurium. AB - Live attenuated Salmonellae may overcome limitations with conventional methods of DNA immunisation. This study examined the impact of plasmid stability on oral DNA delivery by the attenuated Salmonella enterica serovar Typhimurium vaccine strain BRD509. A DNA vaccine cassette comprising the C fragment of tetanus toxin under control of the cytomegalovirus (CMV) promoter was ligated into plasmid pcDNA3, pUC18, pBBR122, pACYC184, pRSF1010/CAT, pBR322 and pAT153. In vitro and in vivo stability studies revealed that, with the exception of pcDNA3 and pUC18, the plasmids were retained by BRD509. However, pAT153 was the only plasmid to induce a tetanus toxoid-specific antibody response following oral delivery. Plasmid copy number was found to impact on plasmid stability and the induction of antigen specific humoral responses. PMID- 17126962 TI - Brief review on the epidemiology of transmissible spongiform encephalopathies (TSE). AB - Transmissible spongiform encephalopathies (TSE) form a group of human and animal diseases that share common features such as (a) distinct pathological lesions in the central nervous system, (b) transmissibility at least in experimental settings, and (c) a long incubation period. Considerable differences exist in the host range of individual TSEs, their routes of transmission, and factors influencing the host susceptibility (such as genotype). The objective of this review was to briefly describe the main epidemiological features of TSEs with emphasis on small ruminant (sheep, goats) TSE, bovine spongiform encephalopathy (BSE) in cattle and chronic wasting disease (CWD) in deer and elk. PMID- 17126963 TI - Safety, immunogenicity and efficacy of modified vaccinia Ankara (MVA) against Dryvax challenge in vaccinia-naive and vaccinia-immune individuals. AB - Modified vaccinia Ankara (MVA) was evaluated as an alternative to Dryvax in vaccinia-naive and vaccinia-immune adult volunteers. Subjects received intramuscular MVA or placebo followed by Dryvax challenge at 3 months. Two or more doses of MVA prior to Dryvax reduced severity of lesion formation, decreased magnitude and duration of viral shedding, and augmented post-Dryvax vaccinia specific CD8(+) T cell responses and extracellular enveloped virus protein specific antibody responses. MVA vaccination is safe and immunogenic and improves the safety and immunogenicity of subsequent Dryvax vaccination supporting the potential for using MVA as a vaccine in the general population to improve immunity to orthopoxviruses. PMID- 17126964 TI - "Blind" testing of models for predicting the 90Sr activity concentration in river systems using post-Chernobyl monitoring data. AB - Two different models for predicting the time-dependent mobility of (90)Sr in river systems have been evaluated using post-Chernobyl monitoring data for five large Belarusian rivers (Dnieper, Pripyat, Sozh, Besed and Iput) in the period between 1990 and 2004. The results of model predictions are shown to be in good agreement (within a factor of 5) with the measurements of (90)Sr activity concentration in river waters over a long period of time after the accident. This verifies the relatively good accuracy of the generalised input parameters of these models which were derived primarily from measurements of (90)Sr deposited after atmospheric nuclear weapons testing (NWT). For the cases studied here, the simpler AQUASCOPE model performed just as well as the more complex "Global" model which used GIS-based catchment data as an input. The reasons for this are discussed. Exponential decay equations were also curve-fitted to the data for each river to help assess the uncertainties in the predictive models. PMID- 17126965 TI - Woman and midwives: position, problems and potential. AB - OBJECTIVE: to describe and analyse birthing models in a number of countries, particularly Samoa and China, that have been the focus of my recent research; to discuss how cultural frameworks, colonisation and ideas of what is 'modern' influence the nature, place of birth and its attendant. IMPLICATIONS FOR PRACTICE: midwives need to reflect on their practice and consider broader health policy and how it affects health systems. They also need to understand the social, economic, historical and cultural context of practice, including the influence of gender inequality and attitudes to women and themselves as midwives. PMID- 17126966 TI - Together or apart? A behavioural and physiological investigation of sleeping arrangements for twin babies. AB - OBJECTIVE: the pros and cons of co-bedding for healthy twin babies on the postnatal ward and in the home are issues with which midwives need to be familiar; however, little objective research has been conducted on which to base policies and recommendations. In this two-part study, I explore the behaviour and physiology of twin babies sleeping separately and together. DESIGN, SETTING AND PARTICIPANTS: in part one, 10 twin baby pairs aged between 1 and 3 months were videoed sleeping together at home in head to head and side by side configurations. In part two, 14 twin pairs under 3 months of age participated in a two-condition trial with behavioural and physiological monitoring while sleeping together and apart in the sleep lab. FINDINGS: in part one, sleep variables were unaffected by co-bedding configuration. Babies positioned side by side were observed to occasionally impinge on, but not obstruct, one another's airways, with an arm across the other's face. In part two, no difference was found in their duration of sleep, frequency of waking, core temperature or head covering in the two conditions; co-bedded twins showed synchronous sleep states. CONCLUSIONS: the co-bedding of term twin babies less than 3 months of age does not seem to be associated with the negative attributes that concern some parents. There may also be advantages of sleep synchrony and ease of care. These results can be used by health professionals in formulating guidance for parents of twins. PMID- 17126967 TI - Breast feeding self-efficacy and other determinants of the duration of breast feeding in a cohort of first-time mothers in Adelaide, Australia. AB - OBJECTIVE: to assess the ability of a Breast-Feeding Self-Efficacy Scale (BSES) score measured at 1 week postpartum to predict the duration of breast-feeding in first-time mothers, and to develop a minimal set of potential confounders, including the BSES and demographic variables, for comparing the apparent effect of other influences on the duration of breast-feeding. DESIGN: a prospective cohort study, with primary outcome the duration of breast feeding up to 6 months postpartum. PARTICIPANTS: 317 women who had given birth to their first baby (at term) in a large teaching maternity hospital in Adelaide, South Australia, during the period March to November, 2003. FINDINGS: the BSES at 1 week postpartum was a strong predictor of the duration of breast-feeding in these first-time mothers. Its ability to predict the duration of breast-feeding was largely independent of the other factors (intended duration of breast-feeding, mother's level of education, country of birth, housing situation, smoking status and method of delivery), which were also found to be significant predictors of breast-feeding duration. IMPLICATIONS FOR PRACTICE: the BSES (including a new short form version) has been confirmed by our study as an important instrument for identifying women at risk of early cessation of breast-feeding. Together with other demographic variables, it should be useful for targeting limited resources to those most in need. PMID- 17126968 TI - Midwives' perception of the intrapartum risk of healthy nulliparae in spontaneous labour, in The Flanders, Belgium. AB - OBJECTIVE: to explore midwives' perception of intrapartum risk for healthy nulliparous women in spontaneous labour at term of a healthy singleton pregnancy, in the Belgian Flanders, and to compare these results with those of a previous study undertaken in England. DESIGN: survey of the care midwives would advocate and their perception of intrapartum risk using a standardised scenario. This study replicates part of a survey undertaken with British midwives (Mead & Kornbrot 2004b). With an added section to capture the particulars of the Belgian situation and explore the likelihood of midwives being fully responsible for the whole intrapartum care of healthy women, including their delivery. The questionnaire was translated into Dutch by MR and distributed by the Flemish Midwives' Association (Vlaamse Organisatie van Vroedvrouwen-VLOV). PARTICIPANTS: all 845 midwives and 143 student midwives who were members of VLOV were sent a questionnaire with their invitation to take part in their annual conference. Two hundred and seventy-five midwives and 107 students attended the conference, and 128 questionnaires were returned at the conference: 99 midwives (36% of the attendees), 26 students (24% of attendees), with three unidentified respondents. This convenience sample represented 12% of all midwives and 18% of all students. ANALYSIS: SPSS for Windows was used for the statistical analysis. Descriptive statistics were used and differences between categorical variables were analysed using chi(2) and Fisher's Exact tests, and differences between continuous variables were analysed by analysis of variance. FINDINGS: midwives generally described a more medicalised approach to intrapartum care on admission and during the first stage of labour than their British counterparts, but were much more optimistic about the chances of healthy women in spontaneous labour achieving a normal delivery within 12 hours. However, Belgian midwives had only a limited ability to undertake normal deliveries because of the high proportion of obstetricians who fulfil this responsibility. This contravenes the European Union (EU) directive on the activities of the midwife. KEY CONCLUSIONS: despite much greater involvement of obstetricians in the care, of women suitable for full midwifery care, and a more medicalised approach to intrapartum care, the Belgian Flanders have a significantly lower caesarean section rate than the UK. The inability of Belgian midwives to fulfil the activities of the midwives as identified by the EU directives raises questions about the migration of midwives trained in Belgium to other EU member states. PMID- 17126969 TI - Relocation of painful end neuromas and scarred nerves from the zone II territory of the hand. AB - This paper reports the results of treatment by proximal relocation of 46 painful end-neuromas or scarred nerves in 33 patients from the pre-defined Zone II of the hand. The relocated nerves included four palmar cutaneous branches of the median nerve, 17 dorsal branches of the ulnar nerves and 25 digital nerves. If no pain at the original site and no pain or only mild pain at the relocation site are considered an adequate treatment of these difficult problems, these relocation procedures achieve complete control of spontaneous baseline pain, complete control of spontaneous spikes of pain, 93% control of direct pressure pain, complete control of movement pain (excluding the extremes of movement of the wrist into extension, supination and, less frequently, pronation) and 96% control of hypersensitivity of the overlying skin. PMID- 17126970 TI - Relationship between recovery of calf-muscle biomechanical properties and gait pattern following surgery for achilles tendon rupture. AB - BACKGROUND: The relationship between ankle plantar flexor biomechanical properties and gait pattern following surgery for acute rupture of the Achilles tendon has not yet been fully investigated. METHODS: Forty-nine young adults (27 men and 22 women) who underwent surgical repair of a complete Achilles tendon rupture were evaluated at 3, 6, 12 and 24 months by clinical assessment, biomechanical evaluation and gait analysis. FINDINGS: Ankle range of motion, plantar flexor passive stiffness and concentric strength were recovered within 12 months. Gait abnormalities related to these factors took longer to disappear owing to the presence of anomalous muscle patterns. At 24 months, a deficit in calf-muscle eccentric strength was still present, determining adaptive changes in gait strategy that involved ankle motion and coordinated muscular activity. INTERPRETATION: Improvement of gait pattern is slower than recovery of plantar flexor mechanical properties. Persisting mechanical impairment resulting in gait adaptations may be detrimental to the healing structures by increasing stress on the Achilles tendon. Restoration of calf-muscle eccentric strength and coordinated antagonist muscle activity should be key points in postoperative rehabilitation following surgical repair of Achilles tendon rupture. PMID- 17126971 TI - Characteristics of the institutionalized and community-residing oldest-old in China. AB - Existing research on the institutionalized population of older adults is primarily limited to Western countries. This study is the first to use nationally representative data to examine differences in the institutionalized and community residing population of the oldest-old (ages 80+) in China. Using three waves of the Chinese Longitudinal Healthy Longevity Survey (CLHLS) (1998, 2000, and 2002), we examine differences in sociodemographic characteristics, family caregiving resources, health practices, religious activity, chronic conditions, and mortality risk. The results indicate that the institutionalized oldest-old are younger, male, reside in urban areas, have lower family-care resources, and exhibit poorer health compared to those living in the community. We also find that the 2-year mortality risk for institutionalized elders is 1.35 times greater than for those residing in the community. However, the mortality differential is eliminated once the sociodemographic, family caregiving, and health characteristics of the oldest-old are taken into account. The implications of these findings are discussed. PMID- 17126972 TI - The poverty-HIV/AIDS nexus in Africa: a livelihood approach. AB - This paper reviews the nexus between poverty and HIV/AIDS in Africa using a sustainable livelihood framework. Much of the literature on HIV and AIDS has generated an almost universal consensus that the AIDS epidemic is having an immense impact on the economies of hard-hit countries, hurting not only individuals, families and firms, but also significantly slowing economic growth and worsening poverty. International evidence has concentrated on the pathways through which HIV/AIDS undermines livelihoods and raises vulnerability to future collapse of livelihoods. Yet, little attention has been paid to the role that social relations and livelihood strategies can play in bringing about risky social interaction that raises the chance of contracting HIV. Using the sustainable livelihood and social relation approaches, this article demonstrates that although AIDS is not simply a disease of the poor, determinants of the epidemic go far beyond individual volition and that some dimensions of being poor increase risk and vulnerability to HIV. PMID- 17126973 TI - Income change at retirement, neighbourhood-based social support, and ischaemic heart disease: results from the prospective cohort study "Men born in 1914". AB - Retirement from active life often leads to decreased finances and reduced social contact, which may increase ischaemic heart disease (IHD) risk in individuals. We examined whether income evolution during the decade before retirement has an impact on subsequent IHD, and explored the mediating effect of common risk factors and social support from different sources (marriage/cohabitation, support from friends/relatives, and neighbourhood-based social support). We analyzed data from the 1982-1983 prospective cohort study, "Men born in 1914" (n=498, follow-up period=10 years) conducted in Malmo, Sweden, merged with yearly income data for 14 years preceding baseline. Low income 10 years before retirement predicted both higher prevalence of IHD risk factors at retirement, and weaker neighbourhood based social support. Income 10 years before retirement was a strong predictor of IHD incidence and mortality after retirement, but a significant downward income mobility at retirement did not increase IHD risk. After adjustment, low neighbourhood-based social support increased the risk of IHD incidence and mortality, and mediated 7-8% of the income effect. In conclusion, income 10 years before retirement, but not the subsequent income evolution, was a strong predictor of IHD post-retirement. This socioeconomic gradient was partly mediated by the protective effect of neighbourhood-based social support, which may be particularly important among the elderly in compensating for social disruptions related to retirement. PMID- 17126974 TI - The effects of olanzapine and fluphenazine on plasma cortisol, prolactin and muscle rigidity in schizophrenic patients: a double blind study. AB - Pharmacotherapy of schizophrenia is associated with the stressful side effects. Muscle rigidity causes distress, discomfort and poor compliance. The aim of the study was to determine the relationship between plasma hormones (cortisol and prolactin/PRL) and muscle rigidity in female schizophrenic patients treated with olanzapine or fluphenazine. In a randomized, double-blind 22-weeks study, 12 patients were treated with olanzapine (5-20 mg/day) and 10 patients received fluphenazine (6-21 mg/day). Treatment with olanzapine moderately decreased, while treatment with fluphenazine significantly increased plasma cortisol levels and muscle rigidity. The marked and moderate increase in plasma PRL levels were found in patients treated with fluphenazine and olanzapine, respectively. The results suggested that olanzapine induced moderate neuroendocrine effects and a reduction in rigidity as compared to fluphenazine treatment. PMID- 17126975 TI - Paralytic ileus associated with combined atypical antipsychotic therapy. AB - First- and second-generation antipsychotics commonly cause mild and sometimes severe gastrointestinal motility depression. We discuss a case of a patient who developed paralytic ileus during his treatment with a combination of second generation antipsychotics. The patient did not receive other medication that could cause depression of intestinal motility than the above-mentioned combination of antipsychotics and no other etiology could be found for the ileus. Furthermore we discuss the theoretical background of antipsychotics induced gastrointestinal motility depression and we provide the literature review of case reports of this topic. PMID- 17126976 TI - Effect of heating on peroxynitrite scavenging capacity of garlic. AB - The ability to scavenge peroxynitrite (ONOO(-)) was studied in the following aqueous garlic extracts: (a) unheated extract of garlic powder (GP), (b) heated extract of garlic powder (HGP), (c) unheated extract of raw garlic (RG), (d) heated extract of raw garlic (HRG), (e) extract of boiled garlic cloves (BG), (f) extract of microwave-treated garlic cloves (MG), and (g) extract of pickled garlic (PG). All the extracts scavenged ONOO(-) in a concentration-dependent way. IC(50) (mg/mL) values for each extract were 0.30+/-0.02 and 0.35+/-0.04 for GP and HGP, respectively; and 0.84+/-0.08, 0.59+/-0.04, 0.76+/-0.09, 1.71+/-0.19, and 1.45+/-0.07 for RG, HRG, BG, MG, and PG, respectively. The ONOO(-) scavenging capacity (IC(50) values) was not decreased in HGP (vs. GP, p>0.05) and in HRG and BG (p>0.05 vs. RG). In contrast, the ONOO(-) scavenging capacity decreased in MG and PG (vs. RG, p<0.001). The IC(50) values for the reference compounds nordihydroguiaretic acid and penicillamine were 1.1 and 4.5mug/mL. The heating before or after garlic cutting was unable to eliminate the capacity of the extracts to scavenge ONOO(-); this capacity was significantly decreased in PG and MG. PMID- 17126977 TI - Ecotoxicological assessment of bromobenzene using a test battery with five model systems. AB - Bromobenzene (BrB) is used as a solvent for crystallization and as an additive to motor oils and may be released into the environment through various waste streams. However, there is limited available information about the toxic hazard of BrB in the aquatic environment. Consequently, the ecotoxicological effects induced by BrB were investigated using five model systems with representants from four trophic levels. The battery included bioluminescence inhibition of the bacterium Vibrio fischeri, growth inhibition of the alga Chlorella vulgaris and immobilization of the cladoceran Daphnia magna. Total protein content, neutral red uptake and MTS metabolization were reduced, while lysosomal function, succinate dehydrogenase activity, G6PDH activity and leakage, metallothionein levels and EROD activity were stimulated in PLHC-1 and RTG-2 fish cell lines. The most sensitive bioindicator was the bioluminiscence of V. fischeri, with an EC(50) of 0.04mM BrB at 15min and a non-observed adverse effect level of 0.02 mM BrB. There is a large difference in sensitivity to BrB among the model systems probably due to the metabolic capacity of the different species. PLHC-1 cells were more sensitive to BrB than RTG-2 cells. The most prominent morphological effects observed were hydropic degeneration, loss of cells and of the perinuclear pattern of distribution of lysosomes. Therefore, BrB should be classified as toxic to aquatic organisms. PMID- 17126978 TI - [Fibular pedicled proximal vascular island flap for distal femoral bone reconstruction. Anatomical study]. AB - The fibular pedicled proximal vascular island flap for distal femoral bone reconstruction described for the first time by Cariou on 1996, is studied in this article with an anatomical view. This anatomical morphometric study allows to assure the surgeon that this procedure can be done and to precise the surgical technique. The authors showed that, dependent on the length of the lower limb and the level of the fibular artery origin, 10 to 19 cm of femoral bone reconstruction could be done, after 180 degrees flap rotation. The flap can be used, the fibula separated in two parts, for inferior femoral metaphysis reconstruction or for knee joint arthrodesis. PMID- 17126979 TI - [Free flaps in elective reconstructive surgery over a 20 year period]. AB - PURPOSE: The purpose of this study was to analyze the evolution of the practice of free flaps in the treatment of tumoral and traumatic pathologies. MATERIALS AND METHODS: This retrospective study was realized on 328 free transfers. The authors compared two periods each of 10 years: series 1 concerns 176 flaps realized from March 12th 1981 to March 13th 1991 (already published); series 2 concerns 152 flaps realized from December 1st 1994 to November 30th 2004. RESULTS: We noticed a decrease of the number of free flaps for lower limbs, a stabilization in the head and neck area and a clear increase in mammary reconstruction. Three donor sites were used in 89% of the cases: latissimus dorsi, forearm and rectus abdominis flaps. Others were used only one to seven times, for specific indications. Some surgical tactics saw their indication decrease (semi-free flap or "apple turnover" technique), others appeared (Y anastomoses). The number of failures decreased from one series to the other (from 5,7 to 2,6%). CONCLUSION: The variety of the flaps remains restricted. The choice is due to the constancy of their vascular anatomy, the size of the pedicles and the reproducibility of their teaching in a University Hospital. PMID- 17126980 TI - [Recurrence of a basal cell carcinoma in a skin graft in spite of total excision. A case report]. AB - The authors report an original case of a recurrence of basal cell carcinoma in a skin graft recipient site. The skin graft was used to resurface the defect following complete excision of basal cell carcinoma in the mandibular angle area. Three answers can be given to the question of the origin of that new carcinoma: recurrence of the primary carcinoma, metastasis to a lymph node or a transfer of a basal cell carcinoma located into the skin graft which grew further. This last hypothesis remains the most probable because the histology of the two carcinomas was different and that the patient presented many others locations of basal cell carcinomas. Skin grafted areas must be checked for recurrence of basal cell carcinoma in such patients because invisible basal cell carcinoma can be transferred within the graft where they can grow for their own. PMID- 17126981 TI - [Evaluation of a method to reduce the number of sentinel nodes removed in melanoma patients: prospective study]. AB - INTRODUCTION: Since 1992, sentinel node (SLN) biopsy was generally applied to melanoma for carcinologic staging. Literature points out an increase of nodes removed for each procedure. It means to a high cost for this procedure and it wanders from the defining concept of SLN. The aim of our study was to evaluate whether, we can minimize number of SLN removed, without influencing the reliability of carcinologic staging. MATERIAL AND METHODS: We conducted a prospective study about 50 patients with stage one melanoma. For each patient, the SLN was identified with hand-held gamma probe technique. We removed only the hottest and all nodes greater than 70% of the hottest. We analysed the characteristics of melanoma, the success rate of this procedure, how many nodes have been removed and how many have had micro-metastases. This result was compared to two main studies with chi(2) test. RESULTS: The success rate of this technique was 100%. We dissected 1,3 SLN for each patient, with 22% positive SLN. Statistical analyse pointed out a better selectivity of our study, rate of pathological positivity and recurrence was alike. DISCUSSION: Our technique decreasing number of removed SLN is reliable. A minimal number of nodes doesn't distort sensitivity of carcinologic staging, and reduce cost of the procedure. PMID- 17126982 TI - Aero-medical evacuation with interventional lung assist in lung failure patients. AB - OBJECTIVE: Acute respiratory failure can make long distance transport by air extremely difficult. Despite pressurised cabins, the pressure will fall to about three quarters of one atmosphere, and the oxygen partial pressure will fall proportionally. Interventional lung assist (iLA) is a well documented treatment in the critical care unit, but has not been evaluated scientifically in long range aero-medical evacuation. The present animal study was performed to test the feasibility of treating lung failure with iLA during intercontinental air evacuation in a military setting. METHODS: Eight adult female pigs were cannulated in the right axillary artery and the right jugular vein. An arterio venous iLA device (Novalung) was connected. The ventilator was adjusted to below half of the needed minute volume before the use of iLA. The animals went through different modalities of transportation in ambulances, helicopters and aircraft. Two of the pigs were tested in a hypobaric chamber, and the remaining two animals underwent a 7.5 h intercontinental transportation from Denmark to Greenland in a Hercules C130J transport airplane. RESULTS: It was possible to maintain physiological PaCO(2) and PaO(2) in normal flight altitudes with iLA. Compared to pump-driven ECMO systems iLA is safer and more efficient. The current study demonstrates the feasibility of iLA during military aero-medical evacuation. PMID- 17126983 TI - Medical and nursing students' attitudes toward cardiopulmonary resuscitation and current practice guidelines. AB - PURPOSE OF THE STUDY: Every member of a healthcare organisation should be able to perform cardiopulmonary resuscitation and defibrillation (CPR-D). The purpose of this cohort study was to examine medical and nursing student's beliefs and attitudes toward CPR-D and current practice guidelines. MATERIALS AND METHODS: A pilot questionnaire concerning beliefs and attitudes toward CPR-D was distributed to 120 fourth year medical students. After statistical analyses (Cronbach's alpha), the questionnaire was modified to increase its reliability. The second version was distributed via e-mail to 100 final sixth year medical students and to 120 final fourth year nursing students. The students had 2 weeks to answer the questionnaire. A reminder was sent via e-mail after 1 week. RESULTS: The questionnaire was answered by 71 of 120 fourth year students (59.1%), 56 of 100 (56.0%) sixth year medical students and 76 of 120 (63.3%) nursing students. Seventy percent of the fourth year, 85.8% of the final (sixth) year medical students and 70.0% of the final (fourth) year nursing students felt confident about their ability to perform basic life support (p<0.01) and 24.0% of the fourth year, 84.0% of the final year medical students and 22.7% of the nursing students about defibrillation (p<0.001). The perceived ability to defibrillate correlated significantly with a positive attitude toward nurse-performed defibrillation (p<0.01) and negatively with fear of damaging the patient's heart by defibrillation (p<0.01). Negative attitude toward defibrillation correlated with perceived organisational attitudes toward practise guidelines (p<0.01). CONCLUSIONS: Medical students' attitudes mature as hoped for, but the nursing students need encouragement. More information is needed to diminish anxiety concerning defibrillation. Negative beliefs and attitudes toward defibrillation affect the students' attitudes toward practice guidelines. PMID- 17126984 TI - Single versus multiple probe blocks of P300-based concealed information tests for self-referring versus incidentally obtained information. AB - The aim of the present study was to compare two protocols and two information types on detection of concealed knowledge. Four independent groups of subjects were run. Two were tested on probe stimuli of a self-referring (AUTO) nature and two were tested on incidentally acquired details of a mock crime (MOCK). Each pair of groups was run either in a one-probe (1PB) or multiple probe (6PB) block. In the single probe block, which was repeated three times with a different probe on each block, one probe item was randomly repeated multiple times in a Bernoulli series with frequently presented, meaningless or irrelevant items. There was also a rare target item designed to force attention to the stimulus screen. All stimulus types were of the same category within each block. In contrast, in the multiple probe (and category) block, rare probes, rare targets and frequent irrelevant items were repeatedly presented in a Bernoulli series within one block. MAJOR RESULTS: There was a difference in task demand as measured by reaction time between the two protocols (the multiple probe protocol was more demanding), and a difference trend in P300 detection sensitivity between protocols for both information types combined in favor of the 1PB (p<.07). With both protocols combined, there was a trend (p<.07) favoring detection of familiar versus incidentally learned information. In terms of P300 amplitudes, both protocols showed the usual result that P300 to probes was greater than that to irrelevants. Also, as with detection rates, self-referring information was better detected in terms of Probe-Irrelevant P300 amplitude differences than mock crime information, regardless of protocol. There was no effect of time passage across the repeated blocks of the one-probe protocol. Methodological problems with the multiple probe protocol as utilized in most recent publications are discussed. PMID- 17126985 TI - Impervious surface as an indicator of pH and specific conductance in the urbanizing coastal zone of New Jersey, USA. AB - Impervious surface is often used as an indicator of non-point source pollution in urban areas due to the strong relationship between percent impervious surface cover and water-quality impacts. In many cases, a threshold effect exists where water quality rapidly degrades above a given percent cover, but the exact threshold level appears to vary across regions. This study explores the relationship between pH, specific conductance, and percent impervious surface cover in the urbanizing coastal zone of New Jersey (USA) to determine the nature of the relationship in the region. The results of the analysis suggest the system is very sensitive to impervious surface, with a threshold potentially existing between 2.4% and 5.1% impervious surface cover. An examination of future conditions suggests that by 2020 water quality in more than 50% of the catchments in the study area will be negatively impacted by non-point source pollution associated with impervious surface. PMID- 17126986 TI - Regional patterns of soil organic carbon stocks in China. AB - Soil organic carbon (SOC) is of great importance in the global carbon cycle. Distribution patterns of SOC in various regions of China constitute a nation-wide baseline for studies on soil carbon changes. This paper presents an integrated and multi-level study on SOC stock patterns of China, and presents baseline SOC stock estimates by great administrative regions, river watersheds, soil type regions and ecosystem. The assignment is done by means of a recently completed 1: 1,000,000 scale soil database of China, which is the most detailed and reliable one in China at the present time. SOC densities of 7292 soil profiles collected across China in the middle of the 1980s were calculated and then linked to corresponding polygons in a digital soil map, resulting in a SOC Density Map of China on a 1: 1,000,000 scale, and a 1 km x 1 km grid map. Corresponding maps of administrative regions, river watersheds, soil types (ST), and ecosystems in China were also prepared with an identical resolution and coordinate control points, allowing GIS analyses. Results show that soils in China cover an area of 9.281 x 10(6)km(2) in total, with a total SOC stock of 89.14 Pg (1 Pg=10(15)g) and a mean SOC density of 96.0 t C/ha. Confidence limits of the SOC stock and density in China are estimated as [89.23 Pg, 89.08 Pg] and [96.143 t C/ha, 95.981 t C/ha] at 95% probability, respectively. The largest total SOC stock (23.60 Pg) is found in South-west China while the highest mean SOC density (181.9 t C/ha) is found in north-east China. The total SOC stock and the mean SOC density in the Yangtze river watershed are 21.05 Pg and 120.0 t C/ha, respectively, while the corresponding figures in the Yellow river watershed are 8.46 Pg and 104.3 t C/ha, respectively. The highest total SOC stocks are found in Inceptisols (34.39 Pg) with SOC density of 102.8 t C/ha. The lowest and highest mean SOC densities are found on Entisols (28.1 t C/ha), and on Histosols (994.728.1 t C/ha), respectively. Finally, the total SOC stock in shrub and forest ecosystem classes are 25.55 and 21.50 Pg, respectively; the highest mean SOC density (209.9 t C/ha) was recorded in the wetland ecosystem class and the lowest (29.0 t C/ha) in the desert ecosystem class. Among five forest ecosystem types, Evergreen conifer forest stores the highest SOC stock (6.81 Pg), and Deciduous conifer forest shows the highest SOC density (225.9 t C/ha). Figures of SOC stocks stratified by Administrative regions, river watersheds, soil types and ecosystem types presented in the study may constitute national-wide baseline for studies of SOC stock changes in various regions in the future. PMID- 17126987 TI - Farm management indicators and farm typologies as a basis for assessments in a changing policy environment. AB - This paper focuses on the assessment of farm management indicators and argues that typologies are a necessary tool for comprehensive environmental assessments. In the paper Andersen et al., [(2004a). Farming and the Environment in the European Community--using agricultural statistics to provide farm management indicators. Paper presented at OECD Expert meeting, March 2004, New Zealand. < http:webdomino1.oecd.org/comnet/agr/farmind.nsf/viewHtml/index/$FILE/Anderson_et_ l.PDF> (1st of February 2006).] presented at the OECD expert meeting on farm management indicators in New Zealand in March 2004, a set of farm management indicators was presented in the framework of a typology of grazing livestock farming systems in the EU-15 (includes all Member States of the European Union before 2004). The present paper presents new results on farm management indicators within the framework of an extended typology for all farming sectors. It presents an environmentally oriented extension to the EU typology of farms currently used for assessing the situation of farming within the European Union. The extended typology is tested in relation to emerging policy issues such as environmental sustainability and rural viability by analysing some of the farm management indicators suggested in Andersen et al., [(2004a). Farming and the Environment in the European Community--using agricultural statistics to provide farm management indicators. Paper presented at OECD Expert meeting, March 2004, New Zealand. (1st of February 2006).]. Finally, recommendations in relation to the next generation EU farm typology are given. PMID- 17126988 TI - The impact of land use and land cover changes on land surface temperature in a karst area of China. AB - Satellite images have been used extensively to study temporal changes in land use and land cover (LULC) in China. However, few studies have been conducted in the karst areas despite the large area and population involved and the fragile ecosystem. In this study, LULC changes were examined in part of Guizhou Province of southern China from 1991 to 2001 based on Landsat Thematic Mapper (TM) images of November 7, 1991, December 5, 1994, and December 19, 2001. Land surface temperature (LST) and normalized difference vegetation index (NDVI) were computed based on LULC types. The results show that agricultural land decreased, while urban areas expanded dramatically, and forest land increased slightly. Barren land increased from 1991 to 1994, and then decreased from 1994 to 2001. These changes in LULC widened the temperature difference between the urban and the rural areas. The change in LST was mainly associated with changes in construction materials in the urban area and in vegetation abundance both in the urban and rural areas. Vegetation had a dual function in the temperatures of different LULC types. While it could ease the warming trend in the urban or built-up areas, it helped to keep other lands warmer in the cold weather. The study also reveals that due to the government's efforts on reforestation, rural ecosystems in some of the study area were being restored. The time required for the karst ecosystem to recover was shorter than previously thought. PMID- 17126989 TI - Vegetation and carbon sequestration and their relation to water resources in an inland river basin of Northwest China. AB - In the Heihe River Basin in the arid inland area of northwest China, the distribution of water resources in vegetation landscape zones controls the ecosystems. The carbon sequestration capacity of vegetation is analyzed in relation to water resources and vegetation growing conditions. During the last 20 years, the vegetation ecosystems have degenerated in the Heihe River Basin. Simulation using the C-FIX model indicates that, at present, the total amount of NPP of vegetation accounts for about 18.16 TgC, and the average value is 106 gC/m(2)/yr over the whole basin. NPP has generally the highest value in the upperstream mountain area, middlestream artificial oases area, downstream river bank area, alluvial fan and the terminal lake depression where vegetation grows relatively well. The lowest value is found in the vast downstream desert and Gobi area. Protection of vegetation ecosystems and enhancement of carbon sequestration require such inland river basins as the Heihe River Basin to be brought under management in a comprehensive way, taking water as a key, to carry out a rational and efficient allocation and utilization of water resources. PMID- 17126990 TI - Greenhouse gas emissions in Canada and Japan: sector-specific estimates and managerial and economic implications. AB - Many firms generate large amounts of carbon dioxide and other greenhouse gases when they burn fossil fuels in their production processes. In addition, production of raw materials and other inputs the firms procure for their operations also generates greenhouse gases indirectly. These direct and indirect greenhouse gas emissions occur in many sectors of our economies. In this paper, we first present sector-specific estimates for such greenhouse gas emissions. We then show that estimates for such sector-specific greenhouse gas emissions are often required for various types of corporate as well as public policy analyses in both domestic and international contexts. Measuring greenhouse gas emissions resulting from firms' multi-stage production processes in a multi-sector context is relevant for policies related to the Kyoto protocol, an international agreement to limit global greenhouse gas emissions. For example, since the protocol allows firms to engage in trading and offsetting of their greenhouse gas emissions across national borders, provided that emissions are correctly measured, the firms can take advantage of such trading schemes by placing their energy-intensive production facilities globally and strategically. We present several case studies which illustrate the importance of this and other aspects of greenhouse gas emissions in firms' environmental management. We also argue that our modeling and estimation methods based on input-output analyses are suitable for the types of research goals we have in this paper. Our methods are applied to data for Canada and Japan in a variety of environmental management circumstances. PMID- 17126991 TI - Sunitinib relieves renal cell carcinoma spinal cord compression. AB - A 66-year-old previously healthy female was diagnosed with renal cell carcinoma (RCC) metastic to the 4th, 6th and 12th thoracic vertebrae. Despite external beam radiotheraphy (20G in 5 fractions), she progressed to spinal cord compression within 4 weeks and was wheelchair bound. Radiation and surgery were not considered suitable, but sunitinib was initiated. After two 6-week sunitinib cycles, she is fully ambulatory. She reports grade I fatigue and occasional epistaxis, as sole side effects. This case demonstrates that despite the rapid progression of the disease, which escaped the effect of radiation within four weeks, sunitinib resulted in complete clinical symptom resolution, which was confirmed radiographically. Moreover, it appears that the novel therapies for metastatic RCC may restore quality and quantity of life to many of those, whose disease previously appeared too advanced to treat. PMID- 17126992 TI - Identification of PEG10 as a progression related biomarker for hepatocellular carcinoma. AB - Widespread DNA copy number alterations are well recognized in hepatocellular carcinoma (HCC), although the affected genes expression remained largely undefined. In this study, we performed genome-wide analysis on HCC to examine the relationship between gene copy number and corresponding transcriptional changes. To ensure analysis on a homogenous population of tumor cells, integrative analysis of array-based CGH and expression profilings was performed on 20 HCC cell lines using a 19,200-element cDNA microarray platform. Further validation studies were carried out on a large series of primary HCC tumors and paired adjacent non-malignant liver to ascertain finding. Correlative analyses highlighted 31 candidate genes that manifested both copy gains and gene up regulations (R2>0.5; p<0.05). Of interest was over-expressed paternally expressed 10 (PEG10) resided within the chromosome region 7q21 that has been implicated in the progression of HCC. Quantitative PCR and qRT-PCR studies verified concurrent genomic gains and over-expression of PEG10 in HCC cell lines and primary tumors (34/40 cases; 85%). In addition, qRT-PCR demonstrated a significant progressive trend of increasing PEG10 expressions from the putative pre-malignant adjacent livers to early resectable HCC tumors, and to late inoperable HCCs (p=0.007). In summary, the present study demonstrated the usefulness of integrated genomic and expression profilings in identifying candidate genes within regions of genomic alteration. Our results also suggested that PEG10 may be a potential biomarker in the progressive development of HCC, and that genomic gain represents one of the major mechanisms in the induction of PEG10 over-expressions. PMID- 17126993 TI - A study of the role of apoptotic cell death and cell cycle events mediating the mechanism of action of 6-hydroxycoumarin-3-carboxylatosilver in human malignant hepatic cells. AB - Previously our research group has studied the anti-proliferative effects of a series of hydroxylated derivatives and silver (I) complexes of coumarin-3 carboxylic acid (C-3-COOH) using two human-derived carcinoma cell lines (A-498 and Hep-G2). Results obtained suggested that both hydroxylation and complexation with silver served to significantly augment the cytotoxic properties of C-3-COOH, to yield a compound, namely 6-hydroxycoumarin-3-carboxylatosilver (6-OH-C-COO-Ag) which could act as a potent and cyto-selective agent, capable of killing cancer cells, and with limited toxicity to cells derived from normal tissue. Here we seek to expand on these findings by probing the molecular mechanism underlying this effect. Results from cytological staining clearly illustrated cellular changes consistent with the induction of apoptotic cell death and which occurred 24 h post-drug-treatment. Additionally, electrophoretic analysis of genomic DNA showed the presence of a ladder pattern, characteristic of apoptotic cell death. This result was subsequently confirmed using a selection of biochemical assays, where increased activity of pro-apoptotic caspases 3 and 9, and increased cleavage of poly(ADP-ribose)-polymerase protein (PARP) were observed. This result was further underpinned by the appearance of a sub-G(1) peak, representing hypo diploid cells, using flow cytometric analysis. Furthermore, 6-OH-C-COO-Ag was seen to function through an alteration in the percentage of cells entering the G(0)/G(1) phase of cell cycle. Consequently, 6-OH-C-COO-Ag has been shown to a more potent and selective anti-cancer agent than cisplatin, capable of altering key biochemical events leading to the execution of apoptotic cell death as early as 24 h post-treatment, suggesting that it may represent a novel therapeutic agent for the safe and effective treatment of cancer in man. PMID- 17126994 TI - Prevalence of human papillomavirus in skin tumors from repair deficient xeroderma pigmentosum patients. AB - The predisposition to skin cancers in childhood is the hallmark of xeroderma pigmentosum (XP), a rare autosomal recessive disorder, deficient in DNA repair and hypersensitive to ultraviolet irradiation. Human papillomavirus (HPVs), are common infections of the skin which are often found associated to benign lesions and non-melanoma skin cancers (NMSC), mainly squamous cell carcinomas (SCC) and basal cell carcinomas (BCC). Our study is the first to analyse 40 SCCs, 27 BCCs and nine normal skin biopsies from XP patients for HPV DNA which are found more frequently in SCCs (20/40) than in BCCs (4/27) or normal skin (2/9). The HPV spectrum includes 22 different epidermodysplasia verruciformis (EV) HPV types, which predominate in SCCs (48%) compared to BCCs (15%) and normal skin (22%). Our data, showing an association between EV HPV and SCCs from young XP patients is comparable to that found for NMSC from adult immunosuppressed organ transplant patients and raises the question of the importance of HPV infection in skin carcinogenesis. PMID- 17126995 TI - BCL-2, TP53 and BAX protein expression in superficial urothelial bladder carcinoma. AB - Whether TP53, BCL-2 and BAX expressions add independent prognostic information in patients with Ta/T1 bladder urothelial carcinoma remains unclear. TP53 overexpression correlated with high tumor grade (p=0.004), WHO grading categories (0.045), BAX expression (p=0.043) and pathologic stage (p=0.05). BCL-2 immunostaining was inverse associated with tumor grade (p=0.008). Lack of BAX expression was related to reduced patient's survival (p=0.028). Mortality was higher in patients with BCL-2+/TP53+ (p=0.023) or TP53+/BAX- (p=0.027) phenotype. BAX and pathologic stage were independent predictors of progression-free and overall survival, respectively. Therefore, BAX expression might be relevant in patient's prognosis. PMID- 17126996 TI - Classification of neuroblastoma patients by published gene-expression markers reveals a low sensitivity for unfavorable courses of MYCN non-amplified disease. AB - Currently, Pubmed lists 385 marker genes for neuroblastoma outcome. Using a customized neuroblastoma-microarray, we evaluated the prognostic impact of the gene-expression pattern of 349 of these candidates (90.6%) in 127 neuroblastoma patients with divergent outcome. By significance analysis of microarrays (SAM) and both uncorrected and Bonferroni-corrected ANOVA, 166/349 (47.5%), 218/349 (62.5%) and 128/349 (36.4%) candidates showed significant differential expression between patients with contrasting outcome. By Prediction Analysis for Microarrays (PAM), a 38-gene-classifier was derived from all markers, which classified patients outcome with an overall accuracy of 78.5%. However, patients with unfavorable outcome of MYCN non-amplified disease were largely misclassified (accuracy: 35%), suggesting that these courses are not identified by current marker genes. PMID- 17126997 TI - Study of quaternary aquifers in Ganga Plain, India: focus on groundwater salinity, fluoride and fluorosis. AB - In marginal and central alluvial plains (Ganga Plain) of India, the inland salinity is continuously increasing, canal network and arid to semi-arid climatic conditions that led to excessive evapotranspiration concentrates the salt in soil and thereby escalating the groundwater salinity. In Mat Tahsil, Mathura district (Ganga Plain) study on shallow and deep aquifer salinity and fluoride was carried out in August 2001 and 2004. Groundwater salinity in some parts is more then 4000 microOmega(-1)/cm. This region is severely affected by endemic fluorosis due to consumption of fluoride-contaminated water. Analysis of F(-), Na(+), K(+), Cl(-) and HCO(3)(-) was carried out at 30 sites of dugwells and borewells. Result shows that there is a variation and continuous escalation in the groundwater salinity and fluoride concentration in deep and shallow aquifers on the basis of analysis. Classification of salinity levels was carried out in 2001 and 2004. The deep aquifers (borewells) are found more saline as compare to the shallow aquifers (dugwells) while F(-), Na(+), K(+), Cl(-) and HCO(3)(-) shows high concentration in shallow aquifers. The fluoride concentration in the groundwater of these villages showed values from 0.1 to 2.5mg/l, severe enough to cause dental and skeletal fluorosis among the inhabitants, especially children of these villages. One of the major effects of inland salinity in this region is from saline groundwater, which is reaching the land surface and causing soil salinisations and water logging in the NE and SE parts of Mat block. PMID- 17126998 TI - Cr(VI) and Cr(III) speciation on Bacillus sphaericus loaded diaion SP-850 resin. AB - A speciation procedure for chromium(III) and chromium(VI) in the environmental samples has been established in the presented work. The procedure presented based on quantitative biosorption of chromium(III) on Bacillus sphaericus loaded Diaion SP-850 at pH 5. The Cr(VI) recoveries at pH 5 were below 5% on the biosorbent. After reduction of Cr(VI) by concentrated H(2)SO(4) and ethanol, the system was applied to the total chromium. Cr(VI) was calculated as the difference between the total chromium content and the Cr(III) content. Optimal analytical conditions including pH, amounts of biosorbent, etc. for Cr(III) recoveries were investigated. The influences of the some alkaline and earth alkaline ions and some transition metals on the recoveries were also investigated. The capacity of biosorbent for chromium(III) was 6.95 mgg(-1). The detection limit (3 sigma) of the reagent blank for chromium(III) was 0.50 microgL(-1). The procedure was successfully applied to the speciation of chromium(III) and chromium(VI) in natural water samples (R.S.D. lower than 5%, recoveries greater than 95%). PMID- 17126999 TI - Chemical release at the airport and lessons learned from the medical perspective. AB - The risk of massive exposure to toxic chemical substances including chemical weapons or industrial chemicals has increased especially during the last century due to the development in industry and chemistry science. This paper aims to describe a real chemical release event and further exposures to personnel working at the Esenboga Airport, Ankara, Turkey, and to give lessons learned. This chemical release was noticed firstly by airport staff giving symptoms including nausea, vomiting, irritation of eyes, itching and rinorrhea. First responders from civil defense unit and a group of health staff including NBC First-aid and Rescue Team gave response to the incident. The increasing number of exposed or suspected cases transferred to hospital were isolated in Emergency Department (ED) following the decontamination at the airport. Due to the characteristic odour and the growing number of the victims, the releasing agent was considered to be likely cyanide or sulfur mustard. Because of the panic amongst the workers, the number of the exposed (real or suspected) people increased up to about 40 and were kept under observation in ED of the hospital. The chromotographic analysis revealed that the agent contained diallyl disulfide, an organo-sulfur compound present at very high concentrations in pure garlic oil. Blood results showed no cyanide and the isolation were terminated. Along with the lessons learned, incident showed that the health facilities should be prepared against such deliberate or accidental mass casualties. PMID- 17127000 TI - Early rapid rise in EAAT2 expression follows the period of maximal seizure susceptibility in human brain. AB - Seizures are relatively common in the first weeks of life and can have lasting effects on brain development due to glutamate excitotoxicity. The excitatory amino acid transporter 2 (EAAT2) is responsible for the majority of glutamate uptake in the brain and mice with this gene deleted die from seizures. Therefore, we reasoned that developmental changes in the expression of EAAT2 might correlate with the period of increased susceptibility to seizures in humans, reflecting a changing vulnerability to excitotoxic insults. Expression levels of eight splice forms of EAAT2 were measured using quantitative RT-PCR from human prefrontal cortex and hippocampus at 1-2 months, 1-2 years, 8 years, 15-16 years, and 18-22 years of age. There was a significant increase in expression of most isoforms between 1-2 months and 1-2 years with isoform-specific patterns after that period. The increase in EAAT2 expression during the first 2 years of life corresponds to a period of maximal synapse formation and other changes in the glutamatergic system such as increased NMDA receptor expression. Moreover, the low expression of EAAT2 in the first months of life corresponds to the period of maximum susceptibility to seizures. PMID- 17127001 TI - PET imaging with [11C]PBR28 can localize and quantify upregulated peripheral benzodiazepine receptors associated with cerebral ischemia in rat. AB - Peripheral benzodiazepine receptors (PBRs) are upregulated on activated microglia. We recently developed a promising positron emission tomography (PET) ligand, [11C]PBR28, with high affinity and excellent ratio of specific to nonspecific binding. We assessed the ability of [11C]PBR28 PET to localize PBRs in a rat permanent middle cerebral artery occlusion (MCAO) model of neuroinflammation. [11C]PBR28 was intravenously administered to rats at 4 and 7 days after permanent MCAO. In all experiments, arterial blood was sampled for compartmental modeling of regional distribution volumes, and rat brains were sampled after imaging for in vitro [3H]PK 11195 autoradiography and histological evaluation. [11C]PBR28 PET and [3H]PK 11195 autoradiography showed similar areas of increased PBRs, especially in the peri-ischemic core. Results from these in vivo and in vitro methods were strongly correlated. In this first study to demonstrate neuroinflammation in vivo with small animal PET, [11C]PBR28 had adequate sensitivity to localize and quantify the associated increase in PBRs. PMID- 17127002 TI - Enhanced excitability of nociceptive trigeminal ganglion neurons by satellite glial cytokine following peripheral inflammation. AB - Peripheral nerve injury activates satellite cells to produce interleukin 1beta (IL-1beta) which mediates inflammation and hyperalgesia. This study investigated the hypothesis that activation of satellite glial cells modulates the excitability of trigeminal ganglion (TRG) neurons via IL-1beta following inflammation. Inflammation was induced by injection of complete Freund's adjuvant (CFA) into the whisker pad area. The threshold for escape from mechanical stimulation applied to the whisker pad in inflamed rats was significantly lower than that in control. Two days post-CFA injection, the mean percentage of TRG neurons encircled by glial fibrillary acidic protein (GFAP)-/IL-1beta immunoreactive cells was significantly increased compared to controls. GFAP and IL-1beta immunoreactivities were coexpressed in the same cells. Fluorogold (FG) labeling identified the site of inflammation. The number of FG-labeled IL receptor type I (IL-1RI) TRG neurons in inflamed rats was significantly greater than in controls. In FG-labeled small TRG neurons, the size of IL-1beta (1 nM) induced-depolarization in inflamed rats was larger than in controls. IL-1beta application significantly increased firing rates evoked by depolarizing pulses in the neurons of inflamed rats, compared to controls. The response to IL-1beta was abolished by treatment with the IL-1RI antagonist. These results suggest that activation of satellite glial cells modulates the excitability of small-diameter TRG neurons via IL-1beta following inflammation, and that the upregulation of IL 1RI in the soma may contribute to the mechanism underlying inflammatory hyperalgesia. Therefore IL-1beta blockers are potential therapeutic agents for prevention of trigeminal hyperalgesia. PMID- 17127004 TI - Identification of Pentatrichomonas hominis in feline fecal samples by polymerase chain reaction assay. AB - Pentatrichomonas hominis is considered to be a commensal protozoan of the vertebrate digestive tract. On the basis of light microscopic examination of feces, some investigators presumptively identified P. hominis as a causative agent of feline diarrhea. However, molecular identification of P. hominis infection in the cat has not been reported. Another trichomonad, Tritrichomonas foetus, is recognized as an intestinal pathogen in cats and often presumptively diagnosed on the basis of the presence of trichomonads in diarrheic feces. It is of importance to determine if cats are natural hosts for P. hominis, as the presence of this organism could result in inaccurate assumption of T. foetus infection. In this study, we used a species-specific PCR assay to identify P. hominis 18S rRNA genes in fecal samples collected from a convenience population of cats in which a high prevalence of T. foetus infection had been previously identified (cat show) or suspected (submitted for T. foetus diagnostic testing). The prevalence of T. foetus infection in these samples was 31% and 28.6%, respectively. P. hominis infection was identified by PCR of DNA extracted from feces of five cats (1.9% and 2.1% of fecal samples, respectively). All cats in which P. hominis was identified were also infected with T. foetus. PCR identification of P. hominis infection in the cat should facilitate future studies to determine the pathogenicity of this species and enable differentiation of P. hominis from other known or as-yet unidentified species of trichomonads that may infect cats. PMID- 17127005 TI - PCR-RFLP for the detection and differentiation of the canine piroplasm species and its use with filter paper-based technologies. AB - Canine piroplasmosis is an emerging disease worldwide, with multiple species of piroplasm now recognised to infect dogs. A nested polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) assay was developed for the detection and differentiation of each of the piroplasm species currently known to infect dogs on the basis of the 18S ribosomal RNA gene. The assay can potentially amplify and discriminate between Theileria annae, Theileria equi, Babesia conradae, Babesia gibsoni, Babesia sp. (Coco) and each of the Babesia canis subspecies. Non-canine piroplasm species can also potentially be detected using the described assay, however amplification of Neospora caninum was also observed. The PCR was found to have a high detection limit, capable of detecting a 2.7x10( 7)% parasitaemia or the equivalent of 1.2 molecules of target DNA when using DNA extracted from whole EDTA blood and detected a parasitaemia of 2.7x10(-5)% using blood applied to both Flinders Technology Associates (FTA) cards and IsoCodetrade mark Stix. The application of blood samples to filter paper may greatly assist in piroplasm identification in regions of the world where local technologies for molecular characterisation are limited. The assay reported here has the potential to be standardised for routine screening of dogs for piroplasmosis. PMID- 17127006 TI - Prevalence and seasonal changes in the population of gastrointestinal nematodes of small ruminants in the semi-arid zone of north-eastern Nigeria. AB - A survey was carried out to determine the prevalence and seasonal abundance of the egg and adult stages of nematode parasites of sheep and goats in the semi arid zone of north-eastern Nigeria between January and December 2002. Faecal samples collected from 102 sheep and 147 goats and examined by the modified McMaster technique using saturated solution of sodium chloride as the floating medium revealed that 44 (43.1%) and 82 (55.8%) of the samples, respectively, contained at least one nematode egg type. Three nematode egg types were recovered with strongyle egg type (22.5% in sheep and 35.4% in goats) being the most prevalent followed, respectively, by Trichuris (5.9% in sheep and 4.1% in goats) and Strongyloides (4.9% in sheep and 4.1% in goats) egg types. Mean faecal egg counts were generally moderate in both sheep (1052+/-922 strongyle, 1000+/-590 Strongyloides and 380+/-110 Trichuris eggs, respectively, per g of faeces) and goats (2092+/-3475 strongyle, 958+/-854 Strongyloides and 683+/-512 Trichuris eggs, respectively, per g of faeces) and showed the same trend irrespective of the age or sex of the animals. The prevalence and counts of strongyle nematode eggs showed a definite seasonal sequence that corresponded with the rainfall pattern in the study area during the period. In both sheep and goats, counts of strongyle egg type increased with the rains and reached peak levels at about the peak of the rainy season in September. The other egg types encountered during the study did not show much variation with the season of the year. Out of the 45 sheep and 75 goats examined at necropsy, 27 (60%) and 39 (52%), respectively, contained adult nematode species. Seven genera of adult nematodes including Strongyloides, Trichostrongylus, Haemonchus, Trichuris, Cooperia, Oesophagostomum and Bunostomum species were encountered during the study. Bunostomum species were recorded only in sheep. Adult worm burdens were generally low and showed seasonal variation that corresponded with the rainfall pattern in the study area during the period. Haemonchus and Trichostrongylus species attained peak counts together in both goats (June) and sheep (August). Strongyloides species were encountered throughout the year in both sheep and goats irrespective of the season. Other genera of nematodes encountered occurred in very low numbers and did not allow any meaningful comparison of seasonal sequence. The results suggest that Haemonchus, Trichostrongylus and Strongyloides species may be the major contributors to small ruminant helminthiasis in the study area. PMID- 17127007 TI - Seasonal allergic dermatitis in sheep in southern Brazil caused by Culicoides insignis (Diptera: Ceratopogonidae). AB - The incidence of seasonal dermatitis was studied in a Hampshire Down flock on a farm in southern Brazil. Epidemiological data, clinical signs and macroscopic pathology were obtained by visiting the farm. Histological lesions were studied in skin biopsies of affected sheep. Biting insects were collected from January to April 2005 in an attempt to identify the etiological agent of the disease. Disease prevalence was 40%; the age of the affected animals was variable. Disease occurred from December to March, some animals had lesions for the entire year. Clinical signs include pruritus on the ears, around the eyes and ventral abdomen. Initially erythema and small red papules were seen, followed by alopecia and crust formation. Histologically the lesions were characterized by perivascular eosinophilic dermatitis. Hyperkeratosis and acanthosis were observed in the chronic lesions. Both Anopheles albitarsis and Culicoides insignis were captured during the study. C. insignis bites caused pruritus in sheep. Both types of insects were caught when they approached the sheep baits approximately 30min after sunset. Results suggested that the disease occurred as a result of an immediate hypersensitivity reaction to C. insignis. PMID- 17127008 TI - Babesia gibsoni rhoptry-associated protein 1 and its potential use as a diagnostic antigen. AB - A cDNA encoding the rhoptry-associated protein 1 (RAP-1) homologue was obtained by immunoscreening an expression library prepared from Babesia gibsoni merozoite mRNA. The complete nucleotide sequence of the gene was 1740bp. Computer analysis suggested that the sequence contains an open reading frame of 1425bp encoding an expected protein with a molecular weight of 52kDa. Based on the sequence similarity, this putative protein was designated as the B. gibsoni RAP-1 (BgRAP 1). The BgRAP-1 gene was expressed in the Escherichia coli BL21 strain, and the recombinant BgRAP-1 was used as the antigen in the enzyme-linked immunosorbent assay (ELISA). The results can differentiate between the B. gibsoni-infected dog sera and the Babesia canis infected dog sera or the normal dog sera. Furthermore, the antibody response against the recombinant protein was maintained during the chronic stage of infection, indicating that the recombinant BgRAP-1 protein might be a useful diagnostic antigen for the detection of antibodies to B. gibsoni infection in dogs. PMID- 17127009 TI - Correlation between the phospholipids domains of the target cell membrane and the extent of Naja kaouthia PLA(2)-induced membrane damage: evidence of distinct catalytic and cytotoxic sites in PLA(2) molecules. AB - Two phospholipase A(2) (PLA(2)) enzymes (NK-PLA(2)-A and NK-PLA(2)-B) were purified from the venom of the monocled cobra Naja kaouthia. The molecular weights of NK-PLA(2)-A and NK-PLA(2)-B, as estimated by mass spectrometry, were 13,619 and 13,303 Da respectively. Both phospholipases were highly thermostable, had maximum catalytic activity at basic pH, and showed preferential hydrolysis of phosphatidylcholine. Intravenous injection of either PLA(2) up to a dose of 10 mg/kg body weight was non-toxic to mice and did not show neurotoxic symptoms. The N. kaouthia PLA(2)s displayed anticoagulant and cytotoxic activity, but poor hemolytic activity. Both the PLA(2)s were more toxic to Sf9 and Tn cells compared to VERO cells. NK-PLA(2) exhibited selective lysis of wild-type baculovirus infected Sf9 cells compared to normal cells. Amino acid modification studies and heating experiments suggest that separate sites in the NK-PLA(2) molecules are responsible for their catalytic, anticoagulant and cytotoxic activities. PMID- 17127010 TI - Several daily measurements are necessary to reliably assess the cortisol rise after awakening: state- and trait components. AB - The cortisol rise after awakening (CAR) is a frequently applied measure of pituitary-adrenal activity. This measure seems to reflect the acrophase of the diurnal cycle and can easily be assessed in saliva samples, collected by the proband or patient under real life conditions. Since different state and trait factors affect the CAR, we here address the questions (a) to which extent state and trait factors affect the CAR, and (b) how often cortisol measures after awakening have to be taken to obtain reliable results. In this study, we assessed the CAR on 6 consecutive days. After applying structural equation models and correlation analyses, we conclude that (a) the CAR of a single day is determined to a great extent by situational factors and only for a small proportion by trait factors and (b) from two (AUC(t)) to six (AUC(i)) days are necessary to achieve reliable trait measures, since state factors bias data from a single day. PMID- 17127011 TI - Edible malaria vaccines that bear fruit. PMID- 17127012 TI - The development of lung damage in asthma is confined to infancy and early childhood. AB - In addition to its symptomatic impact, asthma may be associated with structural lung damage, which is reflected in impairment of lung function. An important and to some extent neglected aspect of the adverse effects of asthma on lung function has been their course: when do they first occur, and how do they progress? For example, does lung function progressively deteriorate, as it does in chronic obstructive pulmonary disease? These questions have in part been addressed by a number of prospective, longitudinal studies of asthma, including the Melbourne Asthma study (Australia), the Dunedin (New Zealand) and the Tucson study (Arizona, USA). Based on these three studies, I propose that the pathological changes which affect lung function occur in infancy and early childhood. The window of opportunity for preventing the development of these changes, and thus modify the course of the disease, is confined to this early period of life. PMID- 17127013 TI - Why there should be more science Nobel prizes and laureates - And why proportionate credit should be awarded to institutions. AB - The four science Nobel prizes (physics, chemistry, medicine/physiology and economics) have performed extremely well as a method of recognizing the highest level of achievement. The prizes exist primarily to honour individuals but also have a very important function in science generally. In particular, the institutions and nations which have educated, nurtured or supported many Nobel laureates can be identified as elite in world science. However, the limited range of subjects and a maximum of 12 laureates per year mean that many major scientific achievements remain un-recognized; and relatively few universities can gather sufficient Nobel-credits to enable a precise estimate of their different levels of quality. I advocate that the Nobel committee should expand the number of Nobel laureates and Prize categories as a service to world science. (1) There is a large surplus of high quality prize candidates deserving of recognition. (2) There has been a vast expansion of research with a proliferation of major sub disciplines in the existing categories. (3) Especially, the massive growth of the bio-medical sciences has created a shortage of Nobel recognition in this area. (4) Whole new fields of major science have emerged. I therefore suggest that the maximum of three laureates per year should always be awarded in the categories of physics, chemistry and economics, even when these prizes are for diverse and un related achievements; that the number of laureates in the 'biology' category of physiology or medicine should be increased to six or preferably nine per year; and that two new Prize categories should be introduced to recognize achievements in mathematics and computing science. Together, these measures could increase the science laureates from a maximum of 12 to a minimum of 24, and increase the range of scientific coverage. In future, the Nobel committee should also officially allocate proportionate credit to institutions for each laureate, and a historical task force could also award institutional credit for past prizes. PMID- 17127014 TI - Mechanism of varenicline (clinical ramifications): electron transfer and oxidative stress. PMID- 17127015 TI - Personal care products that contain estrogens or xenoestrogens may increase breast cancer risk. AB - Established models of breast cancer risk, such as the Gail model, do not account for patterns of the disease in women under the age of 35, especially in African Americans. With the possible exceptions of ionizing radiation or inheriting a known genetic mutation, most of the known risk factors for breast cancer are related to cumulative lifetime exposure to estrogens. Increased risk of breast cancer has been associated with earlier onset of menses or later age at menopause, nulliparity or late first parity, use of hormonal contraceptives or hormone replacement therapy, shorter lactation history, exposure to light at night, obesity, and regular ingestion of alcohol, all of which increase circulating levels of unbound estradiol. Among African Americans at all ages, use of hormone-containing personal care products (PCPs) is more common than among whites, as is premature appearance of secondary sexual characteristics among infants and toddlers. We hypothesize that the use of estrogen and other hormone containing PCPs in young African American women accounts, in part, for their increased risk of breast cancer prior to menopause, by subjecting breast buds to elevated estrogen exposure during critical windows of vulnerability in utero and in early life. These early life and continuing exposures to estrogenic and xenoestrogenic agents may also contribute to the increased lethality of breast cancer in young women in general and in African American women of all ages. Public disclosure by manufacturers of proprietary hormonally active ingredients is required for this research to move forward. PMID- 17127016 TI - Best friends and alcohol consumption in adolescence: a within-family analysis. AB - Although friends and siblings are considered to be important role models in adolescents' peer contexts, these peer influences on adolescent alcohol consumption over time are seldom examined simultaneously in a within-family design. The present study examined the relative impact of alcohol use of the best friend, adolescent sibling and sibling's best friend on the development of alcohol consumption during adolescence. Data reported in this study are part of an ongoing longitudinal questionnaire study among families with two adolescent siblings (N=416). Results from structural equation modeling showed a strong similarity in drinking between best friends and adolescents cross-sectionally. Over time, however, only marginal effects of friends alcohol use on drinking of the youngest sibling, and no effects for the oldest sibling were found. Robust evidence was found for peer-selection processes. In addition, we found a moderate to high relative similarity in drinking within sibling pairs, but no longitudinal effect of sibling's drinking. We also found no support for a possible additional influence of sibling's best friend's drinking on adolescent drinking. Therefore, we tested several potential moderating variables on peer influences, but found no effects of a set of relationship characteristics or individual characteristics on the links between peer and adolescent drinking over time. PMID- 17127017 TI - The epidemiology of fetal alcohol syndrome and partial FAS in a South African community. AB - OBJECTIVES: The prevalence and characteristics of fetal alcohol syndrome (FAS) and partial fetal alcohol syndrome (PFAS) were determined in a third primary school cohort in a community in South Africa (SA). METHODS: An active case ascertainment, two-tier screening methodology, and the revised Institute of Medicine diagnostic criteria were employed among 818 first grade pupils. Characteristics of children with FAS and PFAS are contrasted with a randomly selected control group. Data were collected and analyzed for children in the study regarding: (1) physical growth and development, including dysmorphology, (2) intelligence and behavioral characteristics, and (3) their mother's social, behavioral, and physical characteristics. RESULTS: The rate of FAS and PFAS in this area continues as the highest reported in any overall community and is much higher than rates elsewhere. In this cohort it is 68.0-89.2 per 1000. Severe episodic drinking on weekends among mothers of children with FAS and PFAS accounts for 96% of all alcohol consumed. Various measures of maternal drinking are significantly correlated with negative outcomes of children in the areas of non-verbal intelligence (-0.26), verbal intelligence (-0.28), problem behavior (0.31), and overall dysmorphology score (0.59). Significantly more FAS and PFAS exists among children of rural residents (OR=3.79). CONCLUSIONS: A high rate of FAS and PFAS was again documented in this community, and it has increased. Given population similarities, we suspect that other communities in the Western Cape Province of South Africa also have high rates. Programs for prevention are needed. PMID- 17127018 TI - Anabolic steroid abuse among teenage girls: an illusory problem? AB - BACKGROUND: Recent media reports have portrayed an alarming increase in apparent anabolic-androgenic steroid (AAS) use among American teenage girls; Congress even held hearings on the subject in June 2005. We questioned whether AAS use among teenage girls was as widespread as claimed. METHODS: We reviewed four large national surveys and many smaller surveys examining the prevalence of AAS use among teenage girls. Virtually all of these surveys used anonymous questionnaires. We asked particularly whether the language of survey questions might generate false-positive responses among girls who misinterpreted the term "steroid." We also reviewed data from other countries, together with results from the only recent study (to our knowledge) in which investigators personally interviewed female AAS users. RESULTS: The surveys produced remarkably disparate findings, with the lifetime prevalence of AAS use estimated as high as 7.3% among ninth-grade girls in one study, but only 0.1% among teenage girls in several others. Upon examining the surveys reporting an elevated prevalence, it appeared that most used questions that failed to distinguish between anabolic steroids, corticosteroids, and over-the-counter supplements that respondents might confuse with "steroids." Other features in the phrasing of certain questions also seemed likely to further bias results in favor of false-positive responses. CONCLUSIONS: Many anonymous surveys, using imprecise questions, appear to have greatly overestimated the lifetime prevalence of AAS use among teenage girls; the true lifetime prevalence may well be as low as 0.1%. Future studies can test this impression by using a carefully phrased question regarding AAS use. PMID- 17127019 TI - Passive immunization of guinea pigs with llama single-domain antibody fragments against foot-and-mouth disease. AB - Foot-and-mouth disease (FMD) is a highly contagious disease that occasionally causes outbreaks in Europe. There is a need for therapies that provide rapid protection against FMD in outbreak situations. We aim to provide such rapid protection by passive immunization with llama single-domain antibody fragments (VHHs). Twenty-four VHHs binding serotype O FMDV in vitro were isolated from immunized llamas by phage display and expressed in bakers yeast for further characterization. They recognized four functionally independent antigenic sites. Six strongly FMDV neutralizing VHHs bound to a peptide representing the GH-loop of viral protein 1 known to be involved in binding to the cellular receptor of FMDV. Clone M8, recognizing this antigenic site, and clone M23, recognizing another antigenic site, showed synergistic in vitro virus neutralization. Three FMDV specific VHHs were PEGylated in order to decrease their rapid blood clearance and thus enable in vivo guinea pig protection experiments. Passive immunization with individual VHHs showed no protection, but a mixture of M8 and M23 showed partial transient protection. The protection afforded by these VHHs was however low as compared to the complete protection afforded by convalescent guinea pig serum. In contrast, these VHHs showed far more efficient in vitro FMDV neutralization than convalescent guinea pig serum. This lack of correlation between in vitro neutralization and in vivo protection lends further credence to the notion that opsonophagocytosis of FMDV is important for protection in vivo. PMID- 17127020 TI - Toxicogenomics of endoplasmic reticulum stress inducer tunicamycin in the small intestine and liver of Nrf2 knockout and C57BL/6J mice. AB - This objective of this study was to investigate the toxicogenomics and the spatial regulation of global gene expression profiles elicited by endoplasmic reticulum (ER) stress inducer tunicamycin (TM) in mouse small intestine and liver as well as to identify TM-modulated nuclear factor-E2-related factor 2 (Nrf2) dependent genes. Gene expression profiles were analyzed using 45,000 Affymetrix mouse genome 430 2.0 array and GeneSpring 7.2 software. Microarray results were validated by quantitative real-time reverse transcription-PCR analyses. Clusters of genes that were either induced or suppressed more than two-fold by TM treatment compared with vehicle in C57BL/6J/Nrf2 (-/-; knockout) and C57BL/6J Nrf2 (+/+; wildtype) mice genotypes were identified. Amongst these, in small intestine and liver, 1291 and 750 genes, respectively, were identified as Nrf2 dependent and upregulated, and 1370 and 943 genes, respectively, as Nrf2 dependent and downregulated. Based on their biological functions, these genes can be categorized into molecular chaperones and heat shock proteins, ubiquitination/proteolysis, apoptosis/cell cycle, electron transport, detoxification, cell growth/differentiation, signaling molecules/interacting partners, kinases and phosphatases, transport, biosynthesis/metabolism, nuclear assembly and processing, and genes related to calcium and glucose homeostasis. Phase II detoxification/antioxidant genes as well as putative interacting partners of Nrf2 such as nuclear corepressors and coactivators, were also identified as Nrf2-dependent genes. The identification of TM-regulated and Nrf2 dependent genes in the unfolded protein response to ER stress not only provides potential novel insights into the gestalt biological effects of TM on the toxicogenomics and spatial regulation of global gene expression profiles in cancer pharmacology and toxicology, but also points to the pivotal role of Nrf2 in these biological processes. PMID- 17127021 TI - Sedative and hypnotic activities of the ethanol fraction from Fructus Schisandrae in mice and rats. AB - Fruits of Fructus Schisandrae have been used as medicine for the treatment for insomnia in traditional Chinese medicine. In the present research, the sedative and hypnotic activities of the ethanol fraction of Fructus Schisandrae fruit (SY3) were studied in mice and rats. In the open field test, SY3 (25, 50 and 100 mg/kg) significantly inhibited the motor activity of mice compared to the normal. Results also showed SY3 potentiated pentobarbital-induced sleep by not only increasing the number of falling asleep and prolonging sleeping time but also reducing sleep latency. Furthermore, sleep-wake stages of rats were evaluated by polytrophic recording for 3 h after treatment. The results demonstrated that SY3 at doses of 20, 40 and 80 mg/kg behaved remarkable action on sleep architecture of rats, which contain the increase of total sleeping time, the rate of deep slow wave sleep (SWS) and mean episode duration of deep SWS, and the decrease of the latency of deep SWS. Therefore, these results suggest that the ethanol fraction of Fructus Schisandrae fruit possesses potent sedative and hypnotic activity, which supported its therapeutic use for insomnia. PMID- 17127022 TI - Protective activity of andrographolide and arabinogalactan proteins from Andrographis paniculata Nees. against ethanol-induced toxicity in mice. AB - To find out the active principles against ethanol-induced toxicity in mice, Andrographis paniculata Nees. (Ap) was chosen and isolated andrographolide (ANDRO) and arabinogalactan proteins (AGPs). ANDRO was detected by HPTLC, FTIR and quantified by HPLC (10mg/g of Ap powder). AGPs was detected by beta-glucosyl Yariv staining of SDS-PAGE gel, FTIR and quantified by single radial gel diffusion assay with beta-glucosyl Yariv reagent (0.5mg/g Ap powder). The mice are pretreated intra-peritoneally (i.p.) with different doses (62.5, 125, 250, and 500mg/kg) of body weight of mice] of ANDRO and AGPs for 7 days and then ethanol (7.5g/kg of body weight) was injected, i.p. Besides, silymarin was used as standard hepatoprotective agent for comparative study with ANDRO and AGPs. The ameliorative activity of ANDRO and AGP against hepatic renal alcohol toxicity was measured by assessing GOT, GPT, ACP, ALP and LP levels in liver and kidney. It has been observed that pretreatment of mice with ANDRO and AGPs at 500mg/kg of body weight and 125mg/kg of body weight respectively could able to minimize the toxicity in compare to ethanol treated group as revealed by the different enzymatic assay in liver and kidney tissues and the results were comparable with silymarin. Hence, out of several ill-defined compounds present in Ap, ANDRO and AGPs are the potential bioactive compounds responsible for protection against ethanol-induced toxicity. PMID- 17127023 TI - Contribution of the study of acetabulum for the estimation of adult subjects. AB - INTRODUCTION: The study of the posterior part of the pelvis is of interest in forensic anthropology since it is an anatomical region that is very often preserved. The Lovejoy technique, using the auricular surface has brought about many studies and attracted many authors, among them Buckberry and Chamberlain utilized the criteria described by Lovejoy and developed new methods. By using this research as a starting point, we have described acetabular criteria. The goal of this study was to test these criteria and to link them to auricular surface reading criteria, as described by Buckberry and Chamberlain. METHOD: The study concerned 52 pelvises of known age and sex. We studied the age correlation of various criteria read at the acetabulum level and that of the auricular surface. We then went on to draw up scores and established a correlation of these scores with age. Intra- and inter-observer variability was also studied. We used the SPSS software package for statistical analyses. DISCUSSION: We were able to isolate four acetabular criteria and four auricular surface criteria which have a correlation with age. Establishing these scores (sum of criteria) allows a better age-based correlation to be obtained. Establishing an overall score, including acetabular criteria and auricular surface criteria, allows a good level of correlation to be obtained with age, with low intra- and inter-observer variability. CONCLUSION: The study of acetabulum is of interest for the age estimation of adult subjects. The joint study of the acetabulum and the auricular surface allows a higher correlation with actual age to be obtained. This study must be broadened to include a wider sample so as to allow fine-tuning of a method. PMID- 17127024 TI - Optimization and comparison of three methods for extraction of volatile compounds from Cyperus rotundus evaluated by gas chromatography-mass spectrometry. AB - The essential oil of Cyperus rotundus has multiple pharmacological activities. Therefore, the extraction with high yield and quality is very important for preparation of essential oil of C. rotundus. In this paper, three methods, namely hydrodistillation (HD), pressurized liquid extraction (PLE) and supercritical fluid extraction (SFE), for extraction of volatile compounds from C. rotundus were optimized and compared by gas chromatography-mass spectrometry. Among eight identified compounds in C. rotundus, five components including alpha-copaene, cyperene, beta-selinene, beta-cyperone and alpha-cyperone were quantitatively determined or estimated using alpha-cyperone as standard, which showed that PLE had the highest extraction efficiency, while SFE had the best selectivity for extraction of beta-cyperone and alpha-cyperone. The contents of ingredients from C. rotundus extracted with HD, PLE and SFE are significantly different, which suggest that comparison of chemical components and pharmacological activities of different extracts is helpful to elucidate the active components in C. rotundus and control its quality. PMID- 17127025 TI - Quantitative acid hydrolysis of DE-310, a macromolecular carrier system for the camptothecin analog DX-8951f. AB - DE-310 is a novel macromolecular prodrug of the topoisomerase-I inhibitor DX 8951. DX-8951 is covalently linked to carboxymethyl dextran polyalcohol (CM-Dex PA) via a Gly-Gly-Phe-Gly (GGFG) tetrapeptide spacer. The present study was conducted to identify the portions of DX-8951 linked to DE-310, as well as to quantify the number of DX-8951 molecules associated with DE-310. Two different structures terminated with either glycolaldehyde (CM-GA-GGFG-DX-8951) or glycerol (CM-Glr-GGFG-DX-8951) are obtained when the polymer backbone is fragmented with 1 M HCl. The two products, i.e., CM-GA-GGFG-DX-8951 and CM-Glr-GGFG-DX-8951, indicate linkage of GGFG-DX-8951 with carboxymethyl (CM) group at C-2 and C-4 position of the glucose units, respectively. In the present study, CM-GA-GGFG-DX 8951 was reduced to CM-ethyleneglycol (EG)-GGFG-DX-8951 in order to improve stability prior to HPLC analysis. Hydrolysis results revealed that the amount of CM-GA-GGFG-DX-8951 liberated was 84.7 nmol/mg DE-310 and the amount of CM-Glr GGFG-DX-8951 was 71.8 nmol/mg DE-310. Considering the ratio of generation between CM-GA-GGFG-DX8951 and CM-Glr-GGFG-DX8951, it suggested that slightly larger amount of GGFG-DX-8951 was linked to carboxymethyl groups at the C-2 position of glucose units in DE-310. The sum of the amounts of CM-GA-GGFG-DX-8951 and CM-Glr GGFG-DX-8951 agrees well with the amount of G-DX-8951 produced from DE-310 by alpha-chymotrypsin treatment (157.5 nmol/mg DE-310). The data indicate that the established hydrolysis give a quantitative evaluation of the DX-8951 linked to DE 310. PMID- 17127026 TI - The usefulness of simple X-ray powder diffraction analysis for counterfeit control--the Viagra example. AB - Counterfeit and illegally manufactured drugs become a very important problem all over the world, hence, a search for new fast, easy, reliable and not expensive methods of drugs screening is essential. We describe the use of X-ray powder diffraction method for the fast screening of tablets for counterfeit medicines identification. The original Viagra tablets and some counterfeit and/or imitations of Viagra were used as example. We demonstrate the application of diffraction method for discrimination of tablets coatings and for identification of differences in drug composition. The X-ray diffraction method turns out to be very fast and reliable for detecting counterfeits and imitation, and it correctly predicts the presence or absence of active substance and/or particular excipients. PMID- 17127027 TI - A simple sample preparation with HPLC-UV method for estimation of tiropramide from plasma: application to bioequivalence study. AB - A simple, rapid and selective method was developed for estimation of tiropramide from human plasma. The method involves extracting the tiropramide with n-hexane using diphenhydramine hydrochloride as internal standard. Chromatographic separation was carried out on a reversed phase C(18) column using mixture of water and acetonitrile as mobile phase with UV detection set at 230 nm. The retention time of internal standard and tiropramide were 5.6+/-0.2 and 8.3+/-0.3 min, respectively. The method was validated and found to be linear in the range of 10-200 ng/ml. The co-efficient of variation for intra-day and inter-day accuracy and precision was less than 12.8%. The mean recovery was found to be 89%. An open, randomized, two-treatment, two period, single dose crossover, bioequivalence study in 12 fasting, healthy, male, volunteers was conducted. After dosing, serial blood samples were collected for the period of 12 h. Various pharmacokinetic parameters including AUC(0-t), AUC(0-infinity), C(max), T(max), T(1/2), and elimination rate constant (K(el)) were determined from plasma concentration of both formulations. Log transformed values were compared by analysis of variance (ANOVA) followed by classical 90% confidence interval for C(max), AUC(0-t) and AUC(0-infinity) and was found to be within the range. These results indicated that the analytical method was linear, precise and accurate. Test and reference formulation were found to be bioequivalent. PMID- 17127028 TI - Determination of palonosetron in human plasma by liquid chromatography electrospray ionization-mass spectrometry. AB - A high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) method for the determination of palonosetron (PALO) in human plasma using naloxone as the internal standard (IS) was established. After adjustment to a weakly basic pH with saturated sodium bicarbonate, plasma samples were extracted with ethyl acetate and separated on a Hanbon Lichrospher 5-C18 column with a mobile phase of 40 mM ammonium acetate buffer solution containing 0.04% formic acid-methanol (46:54, v/v). PALO was determined with electrospray ionization-mass spectrometry (ESI-MS). HPLC-ESI-MS was performed in the selected ion monitoring (SIM) mode using target ions at [M+H]+ m/z 297.2 for PALO and [M+H]+ m/z 328.2 for the IS. Calibration curve was linear over the range of 0.02124-10.62 ng/ml. The lower limit of quantification (LLOQ) was 0.02124 ng/ml. The intra- and inter-run variability values were all less than 10.4%. The method has been successfully applied to determine the plasma concentration of PALO in healthy Chinese volunteers. PMID- 17127029 TI - High-performance liquid chromatography analysis of anti-inflammatory pharmaceuticals with ultraviolet and electrospray-mass spectrometry detection in suspected counterfeit homeopathic medicinal products. AB - A simple high-performance liquid chromatography (HPLC) method with both ultraviolet (UV) and electrospray ionisation mass spectrometry (ESI-MS) detection has been developed for the determination of seven pharmaceuticals in counterfeit homeopathic preparations. Naproxen, Ketoprofen, Ibuprofen, Diclofenac, Piroxicam, Nimesulide and Paracetamol were separated by reversed phase chromatography with acetonitrile-water (0.1% acetic acid) mobile phase, and detected by UV at 245 nm and by ESI-MS in negative ionisation mode with the exception of Paracetamol which was detected in positive ionisation mode. Benzoic acid was used as internal standard (IS). This method was successfully applied to the analysis of homeopathic preparations like mother tinctures, solutions, tablets, granules, creams, and suppositories. Linearity was studied with UV detection in the 50-400 microg mL(-1) range and with ESI-MS in the 0.1-50 microg mL(-1) range. Good correlation coefficients were found in both UV and ESI-MS. Detection limits ranged from 0.18 to 41.5 ng in UV and from 0.035 to 1.00 ng in ESI-MS. PMID- 17127030 TI - Single modality and multimodality treatment of nasal and paranasal sinuses cancer: a single institution experience of 229 patients. AB - AIMS: To assess the single and multimodal treatment results and prognostic factors for sinonasal carcinoma. METHODS: Overall survival (OS), disease-specific survival (DSS), local control (LC), and disease-free survival (DFS) in 229 patients with sinonasal carcinoma treated from 1967 to 2003 were calculated. Prognostic factors were univariately and multivariately analyzed. The median follow-up period for survivors was 126 months. RESULTS: 32% of the patients were operated only, 47% underwent multimodal therapy, and 20% were treated without operation. The 5-year OS rate was 41%, and the DSS rate was 51%. The LC rate was 64%, and the DFS rate was 34%. Prognostic for DSS were M status (p<0.001), UICC stage (p<0.001), T classification (p=0.001), N status (p=0.002), intracranial tumor infiltration (p=0.008), infiltration of the pterygopalatine fossa (p=0.02), infiltration of the skull base (p=0.021), infiltration of the orbita (p=0.041), and the type of therapy (p<0.001): The 5-year DSS rate was 63% for patients operated only, 56% for all operated patients, 46% for patients undergoing surgery and radiotherapy, but only 21% for patients treated with radiotherapy+/ chemotherapy. Multivariate analysis revealed that T classification (p=0.042), N classification (p=0.035), M classification (p=0.007), UICC stage (p=0.038), and type of therapy (p=0.038) were independent prognostic factors for DSS. CONCLUSIONS: Radical surgery is recommended for stage I/II sinonasal carcinomas. Stage III/IV carcinomas still have a poor prognosis, but multimodal treatment seems to favor the outcome. PMID- 17127031 TI - Army research needs for automated neuropsychological tests: monitoring soldier health and performance status. AB - Information on the mental status of soldiers operating at the limits of human tolerance will be vital to their management in future deployments; it may also allow earlier intervention for conditions such as undiagnosed Gulf War illnesses and Parkinson's Disease. The Army needs a parsimonious set of neuropsychological tests that reliably identify subtle changes for: (1) early detection of individual health and military performance impairments and (2) management of occupational and deployment health risks. Testing must characterize cognitive lapses in healthy individuals faced with relevant operational stressors (i.e., anxiety, information overload, thermal strain, hypoxia, fatigue, head impact, chemical or radiation exposures, metabolic challenges). This effort must also explore the neuropsychological methods in militarily relevant conditions to extend our understanding of relevant functional domains and how well they correspond to modes of testing. The ultimate objective is unobtrusive real-time mental status monitoring. PMID- 17127032 TI - AICAR potentiates ROS production induced by chronic high glucose: roles of AMPK in pancreatic beta-cell apoptosis. AB - We previously demonstrated that chronic high glucose (33.3 mM) induced beta-cell dysfunction and apoptosis through glucokinase (GCK) downregulation, but the exact mechanisms involved remain unclear. Here, we show that prolonged exposure of 5 aminoimidazole-4-carboxamide (AICA)-riboside potentiated apoptosis induced by high glucose in MIN6N8 pancreatic beta-cells, correlating with enhanced GCK downregulation and decreased production of ATP and insulin. These events are potentiated in AMPK-overexpressing cells, but are prevented in cells transfected with mutant dominant-negative AMPK (AMPK-K45R). Furthermore, AMPK activation increases production of reactive oxygen species (ROS) and loss of mitochondria membrane potential induced by high glucose, which is significantly inhibited by treatment with compound C or by AMPK-K45R overexpression. Overexpression of GCK prevents apoptosis; decreased cellular ATP and insulin secretion, and ROS production enhanced by AICAR, but does not affect AMPK activation. Similar results are obtained using isolated primary islet cells. Collectively, these data demonstrate that AMPK activation potentiates beta-cell apoptosis induced by chronic high glucose through augmented GCK downregulation mediated by enhanced ROS production. PMID- 17127033 TI - Activated stress response pathways within multicellular aggregates utilize an autocrine component. AB - Multicellular aggregates (spheroids) of primary human foreskin fibroblasts (HFF 2) and a glioblastoma cell line (T98G) entered and exited from long term (2 weeks) metabolic arrest utilizing an autocrine response. Cytokine production (specifically IFN-gamma) activated a Gadd45alpha/p38 pathway that led to increased AP-1 (c-jun and ATF3) transcription factor levels, augmenting cytokine production in an autocrine fashion. Whereas HFF-2 aggregates were capable of surviving long term arrest and recovery during NF-kappaB inhibition independent of JNK activation, T98G aggregates were not. Such endogenous processes are not easily observed with adherent monolayer cell culturing systems, strongly suggesting that more emphasis needs to be placed on determining the operational signal transduction cascades within multicellular aggregates. Extracellular inputs such as spheroid formation, arrest, and regrowth as monolayers invoke intracellular signaling responses converging at the AP-1 transcription factor level. Variations in responses are both cell type and transformation state dependent and require an autocrine cytokine component. The data are discussed in relation to the wounding response and avascular tumor growth mechanisms. PMID- 17127035 TI - Shared executive dysfunctions in unaffected relatives of patients with autism and obsessive-compulsive disorder. AB - BACKGROUND: Executive dysfunctions have been studied as a potential endophenotype associated with the genetic basis of autism. Given that recent findings from clinical and molecular genetic studies suggest that autism and obsessive compulsive disorder (OCD) could share a common pattern of heritability, we assessed executive functions as a possible common cognitive endophenotype in unaffected family members of individuals with either autism or OCD. METHODS: Five tests assessing executive functions (Tower of London, verbal fluency, design fluency, trail making and association fluency) were proposed to 58 unaffected first-degree relatives (parents and siblings) of probands with autism and 64 unaffected first-degree relatives of OCD patients. Results were compared with those of 47 healthy controls matched for age, sex, and level of education. RESULTS: In the Tower of London test, both groups of unaffected relatives showed significantly lower scores and longer response times compared with controls. No differences were observed between autism and OCD relatives and healthy controls in the four other tasks (verbal fluency, design fluency, trail making test and association fluency). CONCLUSIONS: Our findings show the existence of executive dysfunction in the unaffected first-degree relatives of probands with OCD, similar to those observed in the relatives of patients with autism. These results support and extend previous cognitive studies on probands indicating executive dysfunctions in autism and OCD. Planning and working memory processes could thus represent a common cognitive endophenotype in autism and OCD that could help in the identification of genes conferring vulnerability to these disorders. PMID- 17127036 TI - Heavy daily smoking among under 18-year-old psychiatric inpatients is associated with increased risk for suicide attempts. AB - PURPOSE: To investigate the relationship between heavy daily smoking and suicidality among adolescent psychiatric inpatients in Finland. METHODS: Data were collected from 411 patients (age 12-17 years) admitted to inpatient psychiatric hospitalization between April 2001 and July 2005. The number of daily cigarettes (>15) and the time of first cigarette after waking up (within 30 min) were used as indicators of heavy daily smoking. RESULTS: After adjusting for psychiatric diagnoses an over twofold risk for suicide attempts was found among adolescents who smoked over 15 cigarettes a day. Additionally, if an adolescent also smoked the first cigarette immediately after waking up the risk was over threefold. Suicidal ideation was not associated with smoking behaviour. DISCUSSION AND CONCLUSIONS: Among adolescents with severe psychiatric illnesses, heavy daily smoking may increase the risk of suicidal behaviour independently of current psychiatric diagnosis. The degree of nicotine dependence of an adolescent should therefore be carefully assessed as part of psychiatric evaluation. PMID- 17127037 TI - Sustained and selective attention deficits as vulnerability markers to psychosis. AB - The first descriptions of schizophrenia emphasized attention problems patients with schizophrenia have but recent results evidence that other psychotic disorders share them. We compared the performance in sustained and selective attention between psychotic patients (P), their healthy first degree relatives (R) and healthy volunteers (C) to prove whether these alterations could be an endophenotype of vulnerability to psychosis. We also compared the performance of schizophrenic patients (SZP) and that of patients with other functional psychoses (OP) in order to prove whether these alterations are specific of any psychotic disorder. Seventy-six P, 70 R and 39 C were included in the study. A selective attention index, comprising TMT A and B and Stroop Test, and a sustained attention index comprising the Continuous Performance Test were calculated. We conducted an univariant general linear model to compare three group performances in these indexes, with age, sex and years of education as a covariables. We found significant differences between the indexes when we compared P, R and C. No differences in performance were found between SZP and OP. Our data showed that sustained and selective attention alterations could be a vulnerability factor to psychotic disorders in general, but they were not specific of schizophrenia. PMID- 17127038 TI - Impaired response inhibition in obsessive compulsive disorder. AB - OBJECTIVE: The present study investigates different three inhibitory control functions in patients with obsessive-compulsive disorder (OCD). Selective motor response inhibition was tested in a GO/NO-GO paradigm, the inhibition of a triggered motor response in a STOP paradigm and the ability to inhibit cognitive interference in a motor STROOP paradigm. METHODS: 27 patients who met DSM-IV criteria for OCD and 25 age, handedness and IQ-matched healthy control subjects were tested in the GO/NO-GO, STOP and motor STROOP tasks. RESULTS: OCD patients performed significantly worse than controls in the selective inhibition of their motor responses (GO/NO-GO) and in the inhibition of cognitive interference (STROOP), and also showed worse performance in suppressing previously triggered motor responses (STOP). CONCLUSION: Patients with OCD are impaired in motor and cognitive inhibitory mechanisms. The findings are consistent with psychobiological and neuropsychological models of OCD suggesting impairment of frontostriatal circuitries that mediate functions of inhibitory control. PMID- 17127039 TI - The role of personality disorder in 'difficult to reach' patients with depression: findings from the ODIN study. AB - Individuals with personality disorders (especially paranoid personality disorder) tend to be reluctant to engage in treatment. This paper aimed to elucidate the role of personality disorder in predicting engagement with psychological treatment for depression. The Outcomes of Depression International Network (ODIN) involves six urban and three rural study sites throughout Europe at which cases of depression were identified through a two-stage community survey. One patient in seven who was offered psychological treatment for depression had a comorbid diagnosis of personality disorder (most commonly paranoid personality disorder). Forty-five percent of patients who were offered psychological treatment for depression did not complete treatment. The odds of completion were higher for patients with a comorbid diagnosis of personality disorder, especially paranoid, anxious or dependent personality disorder. The relatively low number of cases with some specific personality disorders (e.g. schizoid personality disorder) limited the study's power to reach conclusions about these specific disorders. This study focused on a community-based sample which may lead to apparently lower rates of engagement when compared to studies based on treatment-seeking populations. Episodes of depression in the context of personality disorder may represent a valuable opportunity to engage with patients who might otherwise resist engagement. PMID- 17127040 TI - Genetic interaction of Gsc and Dkk1 in head morphogenesis of the mouse. AB - Mouse embryos lacking Gsc and Dkk1 function display severe deficiencies in craniofacial structures which are not found in either Dkk1 homozygous null or Gsc homozygous null mutant embryos. Loss of Gsc has a dosage-related effect on the severity of head truncation phenotype in Dkk1 heterozygous embryos. The synergistic effect of these mutations in enhancing head truncation provides direct evidence of a genetic interaction between Gsc and Dkk1, which display overlapping expression in the prechordal mesoderm. In the absence of Gsc activity, the expression of Dkk1, WNT genes and a transgenic reporter for WNT signalling are altered. Our results show that Gsc and Dkk1 functions are non redundant in the anterior mesendoderm for normal anterior development and Gsc may influence Wnt signalling as a negative regulator. PMID- 17127041 TI - Modulation of CD36 protein expression by AGEs and insulin in aortic VSMCs from diabetic and non-diabetic rats. AB - BACKGROUND AND AIM: In type 2 diabetes, the interplay between cells and inflammatory mediators up-regulates CD36 expression in macrophages. The aim of this work was to investigate advanced glycation end products (AGE)-induced CD36 expression and its regulation by insulin in aortic vascular smooth muscle cells (VSMCs) from Goto-Kakisaki (GK) rats, a non-obese insulin model of both insulin resistance and type 2 diabetes. The context of overexpression of CD36 in aortas was also evaluated. METHODS AND RESULTS: VSMCs were isolated and cultured from the aortas of GK rats and non-diabetic rats. The expression of proteins was evaluated by Western blot. The aortic production of superoxide anion (O(2)(.-)) was measured by luminescence on isolated tissue. AGEs and advanced oxidation protein products (AOPPs) were determined in plasma by fluorescence spectroscopy and spectrophotometry, respectively. AGE receptor (RAGE), NF-kappaB, and CD36 protein expression as well as O(2)(.-) production were higher in GK aortas than in control aortas, and AGEs and AOPPs were higher in GK plasma. In VSMCs from non diabetic rats, insulin was able to reduce (10 nM) or suppress (100 nM) the protein overexpression of CD36 induced by AGEs-BSA. In contrast, in VSMCs from GK rats, insulin was unable to reduce AGEs-BSA-induced CD36 overexpression. CONCLUSIONS: The results suggest an overexpression of CD36 in VSMCs from GK rats and impaired control by insulin. In the context of increased plasma AGEs, aortic RAGE overexpression and increased oxidative stress markers, the data are compatible with an AGEs induced CD36 overexpression in diabetes. PMID- 17127042 TI - Blood glucose and serum lipid levels in the Venezuelan Warao tribe: possible relationship with moriche fruit (Mauritia Flexuosa L.) intake. PMID- 17127043 TI - Fish oil, insulin sensitivity, insulin secretion and glucose tolerance in healthy people: is there any effect of fish oil supplementation in relation to the type of background diet and habitual dietary intake of n-6 and n-3 fatty acids? AB - AIM: To evaluate whether a moderate supplementation of long-chain n-3 fatty acids is able to modulate insulin sensitivity, insulin secretion, beta-cell function and glucose tolerance in healthy individuals consuming a diet rich in either saturated or monounsaturated fat, also in relation to their habitual dietary intake of n-6 and n-3 fatty acid. METHODS AND RESULTS: One hundred and sixty-two healthy individuals were randomly assigned to follow either one of two isoenergetic diets for 3 months, one rich in monounsaturated fats and the other rich in saturated fats. Within each group there was a second randomisation to fish oil (n-3 fatty acids 3.6 g/day) or placebo. At the beginning and at the end of the treatment periods insulin sensitivity (SI), first phase insulin response (FPIR) and glucose tolerance (K(G)-value) were evaluated by the intravenous glucose tolerance test (IVGTT). Fish oil did not have any effect on SI, FPIR, K(G)-value and disposition index in either diet. Even after dividing subjects according to the median value of n-6/n-3 ratio of serum phospholipids at baseline, there was no change in SI (Delta SI 0.42+/-0.34 on fish oil vs 0.14+/ 0.23 on placebo for those with n-6/n-3 <4.85; -1.03+/-0.47 on fish oil vs -0.27+/ 0.32 on placebo for those with n-6/n-3 >4.85) (M+/-SE), FPIR (Delta FPIR 135.9+/ 78.9 vs 157.2+/-157.5 pmol/L; 38.8+/-181.7 vs 357.1+/-181.7 pmol/L), K(G)-value (Delta K(G) 0.14+/-0.15 vs 0.12+/-0.11; -0.32+/-0.16 vs 0.15+/-0.15) or disposition index (Delta disposition index 1465.4+/-830.4 vs 953.8+/-690.0; 1641.6+/-1034.3 vs 446.6+/-905.1). Considering the 75th percentile of n-6/n-3 ratio (5.82) the results on insulin sensitivity, insulin secretion and disposition index were confirmed, while, in this more extreme situation, n-3 fatty acid supplementation induced a significant deterioration of K(G)-value (p=0.02). CONCLUSIONS: In healthy individuals a moderate supplementation of fish oil does not affect insulin sensitivity, insulin secretion, beta-cell function or glucose tolerance. The same is true even when the habitual dietary intake of n-6 and n-3 fatty acids is taken into account. PMID- 17127044 TI - Dietary fiber and control of body weight. PMID- 17127045 TI - Development and bioavailability assessment of ramipril nanoemulsion formulation. AB - The objective of our investigation was to design a thermodynamically stable and dilutable nanoemulsion formulation of Ramipril, with minimum surfactant concentration that could improve its solubility, stability and oral bioavailability. Formulations were taken from the o/w nanoemulsion region of phase diagrams, which were subjected to thermodynamic stability and dispersibility tests. The composition of optimized formulation was Sefsol 218 (20% w/w), Tween 80 (18% w/w), Carbitol (18% w/w) and standard buffer solution pH 5 (44% w/w) as oil, surfactant, cosurfactant and aqueous phase, respectively, containing 5 mg of ramipril showing drug release (95%), droplet size (80.9 nm), polydispersity (0.271), viscosity (10.68 cP), and infinite dilution capability. In vitro drug release of the nanoemulsion formulations was highly significant (p<0.01) as compared to marketed capsule formulation and drug suspension. The relative bioavailability of ramipril nanoemulsion to that of conventional capsule form was found to be 229.62% whereas to that of drug suspension was 539.49%. The present study revealed that ramipril nanoemulsion could be used as a liquid formulation for pediatric and geriatric patients and can be formulated as self nanoemulsifying drug delivery system (SNEDDS) as a unit dosage form. PMID- 17127046 TI - Copper impairs biliary epithelial cells and induces protein oxidation and oxidative DNA damage in the isolated perfused rat liver. AB - Copper is one of the major metals causing environmental contamination. Previous studies showed that copper induced toxic effects in isolated perfused rat liver models and these effects were associated with lipid peroxidation. Here we investigated whether effects of copper (at concentrations of 0.01, 0.03, and 0.1 mM of Cu(2+) in Krebs-Henseleit buffer perfusing the isolated rat liver for 60 min), were associated with biliary epithelial cell injury, as well as protein oxidation and oxidative DNA damage. The highest concentration of copper in perfusate (0.1 mM) did not allow complete evaluation of all parameters because it blocked portal flow within 30 min of perfusion, indicating severe microcirculatory disturbances. Further, copper decreased secretion of bile and it increased lactate dehydrogenase, aspartate transaminase, and alanine transaminase leakage into perfusate as well as liver weight in a dose-dependent manner. Biliary gamma-glutamyltransferase, an index of biliary epithelial cell integrity increased similarly at 0.01 and 0.03 mM copper concentrations in perfusate. Compared to controls, 0.01 and 0.03 mM concentrations of copper increased the amount of thiobarbituric acid reacting substances, a marker of lipid peroxidation, tissue protein carbonyl groups, an index of protein oxidation, and 8-oxo-7,8-dihydro-2'-deoxyguanosine, a marker of oxidative DNA damage. The results suggest that toxic effects of copper in the isolated perfused rat liver may involve biliary epithelial cells and they are associated with lipid peroxidation, protein oxidation, and oxidative DNA damage. PMID- 17127047 TI - Fungal antagonists of the plant pathogen Rhizoctonia solani: selection, control efficacy and influence on the indigenous microbial community. AB - A broad spectrum of fungal antagonists was evaluated as potential biocontrol agents (BCAs) against the soil-borne pathogen Rhizoctonia solani using a new combination of in vitro and in vivo assays. The in vitro characterisation of diverse parameters including the ability to parasitise mycelium and to inhibit the germination of Rhizoctonia sclerotia at different temperatures resulted in the selection of six potential fungal antagonists. These were genotypically characterised by their BOX-PCR fingerprints, and identified as Trichoderma reesei and T. viride by partial 18S rDNA sequencing. When potato sprouts were treated with Trichoderma, all isolates significantly reduced the incidence of Rhizoctonia symptoms. Evaluated under growth chamber conditions, the selected Trichoderma isolates either partly or completely controlled the dry mass loss of lettuce caused by R. solani. Furthermore, the antagonistic Trichoderma strains were active under field conditions. To analyse the effect of Trichoderma treatment on indigenous root-associated microbial communities, we performed a DNA-dependent SSCP (Single-Strand Conformation Polymorphism) analysis of 16S rDNA/ITS sequences. In this first assessment study for Trichoderma it was shown that the pathogen and the vegetation time had much more influence on the composition of the microbiota than the BCA treatment. After evaluation of all results, three Trichoderma strains originally isolated from Rhizoctonia sclerotia were selected as promising BCAs. PMID- 17127048 TI - IL-12-induced T-bet expression and IFNgamma release in lymphocytes from asthmatics--role of MAPkinases ERK-1/-2, p38(MAPK) and effect of dexamethasone. AB - The transcription factor T-box-expressed-in-T-cells (T-bet) is required for T(H)1 lymphocyte differentiation, regulates the IL-12-induced expression of the T(H)1 specific cytokine IFNgamma and may be dysregulated in asthmatics. The modulatory role of extracellular signal-regulated kinase (ERK)-1/-2, p38mitogen-activated protein kinase (MAPK) and dexamethasone on IL-12 induced T-bet and IFNgamma expression was assessed in peripheral blood lymphocytes of 10 atopic asthmatics and 10 nonatopic normals. IFNgamma production was dependent on phosphorylation of ERK-1/-2 and p38MAPK, as examined by PD098059, an inhibitor of the upstream activator of MAPKkinase (MKK-1), and SB203580, an inhibitor of p38MAPK. The inhibitory effect of PD098059 on IFNgamma release was decreased in asthmatic T cells compared with normals. The IL-12-induced T-bet expression and the inhibitory effect of SB203580 were increased in asthmatic T-cells compared with normals. Dexamethasone blocked the IL-12-induced T-bet expression in asthmatic T cells completely and decreased IL-12-induced IFNgamma release by approximately 50%, which occurred to the same extent in asthmatic and normal T-cells. In conclusion, (1) p38MAPK-pathway rather than ERK-pathway may play a more basic role in the regulation of the increased T-bet expression in asthma, and (2) ERK- and p38MAPK-activation modulate IFNgamma expression independently of T-bet and this regulatory role of ERK-1/-2 on IFNgamma release is impaired in asthma. The therapeutic benefit of dexamethasone on T-bet and IFNgamma production seems to be critical. PMID- 17127049 TI - Snoring and daytime sleepiness as risk factors for hypertension and diabetes in women--a population-based study. AB - The aim of this study was to analyze whether snoring and excessive daytime sleepiness (EDS), the main symptoms of obstructive sleep apnea syndrome (OSAS), are associated with hypertension and diabetes in women. A random sample of 6779 women aged 20-99 years answered questionnaires on sleep disturbances, daytime symptoms and somatic diseases. The women were categorized into four groups: "no EDS or snoring" (reference group), "snoring but no EDS", "EDS but no snoring" and "snoring and EDS". Prevalences of hypertension and diabetes were lowest in the reference group (8.7% and 1.6%, respectively) and highest among women with both snoring and EDS (hypertension: 26.3%, diabetes: 5.8%). In a multivariate model adjusting for age, body mass index, smoking, physical activity and alcohol dependency, "snoring and EDS" was a risk factor for hypertension (adjusted OR 1.82 (95% CI 1.30-2.55)) while isolated snoring or EDS was not. "Snoring and EDS" was more closely related to hypertension among women aged <50 years (adj. OR 3.41 (1.78-6.54) vs. 1.50 (1.02-2.19), P=0.01). For diabetes, both "EDS but no snoring" and "snoring and EDS" were risk factors and the associations were most pronounced in women aged >50 years (adj. OR 2.33 (1.28-4.26) for "EDS but no snoring" and 2.00 (1.05-3.84) for "snoring and EDS"). We conclude that the combination of snoring and EDS is a risk factor for hypertension and diabetes in women. For hypertension, the risk is partly age dependent and, for diabetes, EDS without snoring is a risk factor of similar magnitude. These differences might indicate differences in pathophysiologic mechanisms underlying the association between sleep-disordered breathing and hypertension and diabetes respectively. PMID- 17127050 TI - Differential proteomic analysis of the anti-proliferative effect of glucocorticoid hormones in ST1 rat glioma cells. AB - Glucocorticoid hormones (GCs) exert a potent anti-proliferative activity on several cell types. The classic molecular mechanism of GCs involves modulation of the activity of the glucocorticoids receptor, a transcriptional regulator. However, the anti-proliferative effect of GCs may also involve modulation of processes such as translation, subcellular localization and post-translational modifications, which are not reflected at the mRNA level. To investigate these potential effects of GCs, we employed the proteomic approach (two-dimensional electrophoresis and mass spectrometry) and the ST1 cells, obtained from the C6 rat glioma cell line, as a model. GC treatment leads ST1 cells to a complete transformed-to-normal phenotypic reversion and loss of their tumorigenic potential. By comparing sets of 2D nuclear protein profiles of ST1 cells treated (or not) with hydrocortisone (Hy), 13 polypeptides displaying >or=two-fold difference in abundance upon Hy treatment were found. Five of these polypeptides were identified by peptide mass fingerprinting, including Annexin 2 (ANX2), hnRNP A3 and Ubiquitin. Evidence obtained by Western blot analysis indicates that ANX2 is present in the nucleus and has its subcellular localization modulated by GC treatment of ST1 cells. Our findings indicate complementary mechanisms contributing to the regulation of gene expression associated with ST1 cells' response to GCs. PMID- 17127051 TI - Expression and characterization of Acidothermus cellulolyticus E1 endoglucanase in transgenic duckweed Lemna minor 8627. AB - Endoglucanase E1 from Acidothermus cellulolyticus was expressed cytosolically under control of the cauliflower mosaic virus 35S promoter in transgenic duckweed, Lemna minor 8627 without any obvious observable phenotypic effects on morphology or rate of growth. The recombinant enzyme co-migrated with the purified catalytic domain fraction of the native E1 protein on western blot analysis, revealing that the cellulose-binding domain was cleaved near or in the linker region. The duckweed-expressed enzyme was biologically active and the expression level was up to 0.24% of total soluble protein. The endoglucanase activity with carboxymethylcellulose averaged 0.2 units mg protein(-1) extracted from fresh duckweed. The optimal temperature and pH for E1 enzyme activity were about 80 degrees C and pH 5, respectively. While extraction with HEPES (N-[2 hydroxyethyl]piperazine-N'-[2-ethanesulfonic acid]) buffer (pH 8) resulted in the highest recovery of total soluble proteins and E1 enzyme, extraction with citrate buffer (pH 4.8) at 65 degrees C enriched relative amounts of E1 enzyme in the extract. This study demonstrates that duckweed may offer new options for the expression of cellulolytic enzymes in transgenic plants. PMID- 17127052 TI - Production of ethanol from starch by free and immobilized Candida tropicalis in the presence of alpha-amylase. AB - Candida tropicalis is a potentially useful organism for the commercial production of ethanol as it is capable of fermenting starch at a low rate. To enhance this carbon source utilization and increase the rate of alcohol production, we pretreated corn soluble starch with alpha-amylase. Starch liquefaction was sufficient to drive the fermentation and to convert 96% substrate to ethanol. Indeed, in the presence of exogenous alpha-amylase, 9% (w/v) soluble starch was converted to 43.1g ethanol/l in 65 h with a productivity of 0.65 g/l h. Thus, bio ethanol production using free and calcium alginate-immobilized C. tropicalis does not require the saccharification step. Furthermore, fed-batch fermentation by free C. tropicalis cells increased the final concentration to 56 g ethanol/l, reaching published values for Saccharomyces cerevisiae recombinant strains expressing both alpha-amylase and glucoamylase. PMID- 17127053 TI - Evaluation by respirometry of the loading capacity of a high rate vermicompost bed for treating sewage sludge. AB - This study examines high rate vermicomposting of sewage sludge using high stocking densities of earthworms. To examine the loading capacity, two vermicompost beds were established in identical 0.84 m diameter reactors, one loaded at an average rate of 10 kg-wet-sludge-mixture m(-2) day(-1) (0.5 kg carbon m(-2) day(-1)), the other loaded at 20 kg-wet-sludge-mixture m(-2) day(-1) (1 kg-carbon m(-2) day(-1)). The sludge mixture was from a commercial vermicomposting company (Vermitech) and contained 80-90% sludge and 10-20% green waste and clay. The beds were operated in fed-batch mode for 38 days, and then monitored for a further 12 days without any further sludge loading. Earthworms (Eisenia fetida) were added once or twice weekly over the 38 days loading period to gradually decrease the ratio of sludge loading rate to total earthworm biomass. By adding earthworm incrementally, the feeding rate ranged from 3.9 to 1.25 kg-wet-sludge kg-earthworm(-1) day(-1) for the full load experiment and 2 0.62 kg-wet-sludge kg-earthworm(-1) day(-1) for the half load experiment. The extent of degradation was estimated by fitting a 1st order model to the CO2 production rate from the beds. Based on the 1st order model, 53+/-20% (95% CI) and 68+/-4% of the organic carbon was converted to CO2 -C in the full load and half load experiments respectively. The CO2 production rate in the half load experiment became stable and repeatable when the total earthworm biomass reached 5.4 kg, corresponding to a feed rate of 1.04 kg-wet-sludge-mixture kg-earthworm( 1) day(-1). In contrast, the rate of CO2 production was still climbing and traces of methane were evident in the full load experiment at the end of the 38 day loading period. The experiments indicate that high rate vermicomposting beds are sustainable providing the feeding rate does not exceed approximately 1 kg-wet sludge kg-earthworm(-1) day(-1). PMID- 17127054 TI - Amplification of DNA-binding affinities of protoberberine alkaloids by appended polyamines. AB - This communication describes a synthetic approach toward the amplification of the moderate DNA-binding affinities of protoberberine alkaloids. Specifically, three protoberberine derivatives bearing two to six primary amino groups at the 3- and 9-positions of protoberberine were synthesized and characterized by NMR ((1)H and (13)C) and HRMS. Studies on their affinities toward calf thymus (CT) DNA by ethidium bromide (EB) displacement and spectrophotometric titration experiments indicate that these polyamino protoberberines show more than 10(3)-fold enhanced DNA-binding affinities relative to palmatine and thus are exploitable as strong DNA-binders. PMID- 17127055 TI - Novel carbazole derivatives as NPY Y1 antagonists. AB - The synthesis of a series of carbazole derivatives and their SAR at the NPY Y1 receptor is described. Modulation of physicochemical properties by appropriate decoration led to the identification of a high-affinity NPY Y1 antagonist that shows high brain penetration and modest oral bioavailability. PMID- 17127056 TI - A convenient and efficient protocol for oxidative aromatization of Hantzsch 1,4 dihydropyridines using benzyltriphenylphosphonium peroxymonosulfate under almost neutral reaction conditions. AB - Oxidative aromatization of 4-alkyl or aryl and heterocyclic-substituted derivatives of Hantzsch 1,4-dihydropyridines to the corresponding pyridine derivatives has been studied using benzyltriphenylphosphonium peroxymonosulfate as an oxidant in the presence of BiCl(3) under nearly neutral reaction conditions at ambient temperature. PMID- 17127057 TI - Carbonic anhydrase activators: L-Adrenaline plugs the active site entrance of isozyme II, activating better isoforms I, IV, VA, VII, and XIV. AB - The activation of the metalloenzyme carbonic anhydrase (CA, EC 4.2.1.1) with L adrenaline and histamine has been investigated by kinetic and X-ray crystallographic studies. L-Adrenaline behaves as a potent activator of isozyme CA I (activation constant of 90 nM), being a much weaker activator of isozyme CA II (activation constant of 96 microM). Isoforms CA IV, VA, VII, and XIV were activated by L-adrenaline with K(A)s in the range of 36-63 microM. The X-ray crystal structure of the CA II-L-adrenaline adduct revealed that the activator plugs the entrance of the active site cavity, obstructing it almost completely. PMID- 17127058 TI - Synthesis and in vitro photodynamic activity of mono-substituted amphiphilic zinc(II) phthalocyanines. AB - A series of novel zinc(II) phthalocyanines mono-substituted with a 1,3 bis(dimethylamino)-2-propoxy group at the alpha- or beta-position, and the corresponding di-N-methylated derivatives, have been synthesized. All these compounds can generate singlet oxygen effectively and exhibit high in vitro photodynamic activities toward HT29 human colorectal carcinoma cells with IC(50) values down to 0.08microM. The dicationic derivatives have a higher affinity to the cell membrane compared with the non-ionic counterparts. PMID- 17127059 TI - Novel tetrahydroisoquinolines are histamine H3 antagonists and serotonin reuptake inhibitors. AB - A series of novel 4-aryl-1,2,3,4-tetrahydroisoquinoline-based histamine H(3) ligands that also have serotonin reuptake transporter inhibitor activity is described. The synthesis, in vitro biological data, and select pharmacokinetic data for these novel compounds are discussed. PMID- 17127060 TI - Synthesis of OSW saponin analogs with modified sugar residues and their antiproliferative activities. AB - Eight monosaccharide analogs of the potent antitumor OSW saponins (2-9) were synthesized. One analog, 2-O-acetyl-alpha-l-arabinopyranoside 3, showed antiproliferative activity against the Jurkat cells (IC(50)=0.078microM) comparable to that of the disaccharide derivative (1). PMID- 17127061 TI - Synthesis and cytotoxic activities of 4,5-diarylisoxazoles. AB - A series of 4,5-diarylisoxazoles related to combretastatin A-4 (CA-4) were synthesized and evaluated for cytotoxicity against three human cancer cell lines. Among them, compound 6e showed better cytotoxic activity than CA-4 in HeLa and HepG2 cell lines assayed with IC(50) value as low as 0.022 and 0.065nM, respectively. PMID- 17127062 TI - 1,3-Disubstituted-imidazo[1,5-a]pyrazines as insulin-like growth-factor-I receptor (IGF-IR) inhibitors. AB - A series of novel 8-amino-1,3-disubstituted-imidazo[1,5-a]pyrazines was designed and synthesized as IGF-IR inhibitors. PMID- 17127063 TI - Carbonic anhydrase inhibitors. Inhibition studies of the human secretory isoform VI with anions. AB - The unique secretory isozyme of human carbonic anhydrase (hCA, EC 4.2.1.1), hCA VI, has been cloned, expressed, and purified. The kinetic parameters for the CO(2) hydration reaction proved hCA VI to possess a k(cat) of 3.4x10(5)s(-1) and k(cat)/K(M) of 4.9x10(7)M(-1)s(-1) (at pH 7.5 and 20 degrees C). hCA VI has a significant catalytic activity for the physiological reaction, of the same order of magnitude as isoforms CA I or CA IX. A series of anions (such as bicarbonate, chloride, nitrate, etc.) were shown to inhibit the activity of the enzyme, with inhibition constants typically in the range of 0.60-0.90mM. The best hCA VI inhibitors were cyanide, azide, sulfamide, and sulfamate, with inhibition constants in the range of 70-90microM. PMID- 17127064 TI - Intra-articular synovial lipoma of the knee joint (located in the lateral recess): a case report and review of the literature. AB - This report describes a rare intra-articular synovial lipoma of the knee joint which developed in a 66-year-old female. The patient suffered from sudden knee pain and a catching or slight interruption of normal motion of the left knee and then noticed the tumor. The tumor was located in the lateral recess of the knee joint and showed a signal intensity similar to subcutaneous fat on T1 and T2() weighted magnetic resonance images. The arthroscopy revealed a smooth, globular, yellowish, encapsulated tumor extending into the lateral recess from the surface of the lateral condyle and the tumor was totally excised under arthroscopic guidance. Histologic examination of the specimen revealed a tumor composed of mature adipose cells covered by a thin fibrous layer of varying thickness and normal synovial lining cells. The diagnosis was intra-articular synovial lipoma. Intra-articular synovial lipomas should be distinguished from other similar lipomatoid conditions such as Hoffa disease and villous lipomatous proliferation of the synovial membrane (lipoma arborescens). Intra-articular synovial lipoma should be considered in the differential diagnoses when examining a patient with sudden knee pain, and a catching or locking knee. PMID- 17127065 TI - A prospective study of the diagnostic potential of the knee tunnel view radiograph in assessing anterior knee pain. AB - The aim of this comparative study was to examine the potential advantage of the tunnel view radiograph over a series of weight bearing antero-posterior (AP), lateral and skyline radiographs. The study population consisted of 240 subjects with knee pain aged 19 to 93 years. A total of 309 knees had a weight bearing AP in extension, lateral, skyline and tunnel view radiographs. Each radiograph was reported with respect to features related to osteoarthritis, modified from the Ahlback system. Each feature was assessed using the tunnel radiograph alone and then the AP, lateral and skyline views in combination without, and blind to, the information from the tunnel view. On the basis of Bowker's test, the tunnel view was more likely to pick up abnormal intercondylar notch and tibial spine osteophytes but not loose bodies. We conclude that the tunnel view is a valuable addition in the routine assessment of the knee joint in osteoarthritis but not for the diagnosis of loose bodies alone. PMID- 17127066 TI - Design and synthesis of piperidine farnesyltransferase inhibitors with reduced glucuronidation potential. AB - The design and synthesis of a novel piperidine series of farnesyltransferase (FTase) inhibitors with reduced potential for metabolic glucuronidation are described. The various substitution and exchange of the phenyl group at the C-2 position of the previously described 2-(4-hydroxy)phenyl-3-nitropiperidine 1a (FTase IC(50)=5.4nM) resulted in metabolically stable compounds with potent FTase inhibition (14a IC(50)=4.3nM, 20a IC(50)=3.0nM, and 50a IC(50)=16nM). Molecular modeling studies of these compounds complexed with FTase and farnesyl pyrophosphate are also described. PMID- 17127067 TI - Methotrexate gamma-hydroxamate derivatives as potential dual target antitumor drugs. AB - A series of new aminopteroyl-based hydroxamate derivatives were synthesized and tested in vitro in cell culture models as potential dual target drugs. These compounds were designed to target two families of enzymes, matrix metalloproteinases (MMP) and a folate enzyme, dihydrofolate reductase (DHFR). These enzymes are the components of two unrelated cellular pathways and they are often over-expressed in metastasizing tumors. In addition to the synthesis and full structural characterization of the hybrid molecules, we describe their inhibitory activities against a series of MMPs (MMP-2, MMP-7, MMP-9, MMP-14) and DHFR, as well as their antiproliferative activity in three cancer cell lines. The new hydroxamate derivatives of MTX proved to be effective inhibitors of MMPs and DHFR in the micromolar and nanomolar range, respectively. Furthermore, they showed strong antiproliferative activity against A549 cells (non-small cell lung carcinoma), and PPC-1 and Tsu-Pr1 prostate cancer cell lines. Therefore, based on the present results, these bi-functional drugs may be good candidates to target specific tumors in animal models due to potential combined effects on two pathways crucial for tumor development. PMID- 17127068 TI - Chiral multinuclear macrocyclic polyamine complexes: synthesis, characterization and their interaction with plasmid DNA. AB - A series of multinuclear macrocyclic polyamine metal (Zn(2+), Cu(2+), Co(2+)) complexes containing chiral dipeptide linkage were synthesized and used as artificial nuclease enzyme model. The interaction between the complexes and plasmid DNA (pUC19) was studied, and the results revealed that these complexes could act as powerful catalysts for the cleavage of plasmid DNA under physiological conditions. PMID- 17127070 TI - Synthesis and evaluation of unsaturated caprolactams as interleukin-1beta converting enzyme (ICE) inhibitors. AB - Peptidomimetic compounds possessing a caprolactam ring constraint were prepared and evaluated as interleukin-1beta converting enzyme (ICE) inhibitors. The caprolactam ring was used to constrain the P3 region of our inhibitors. This strategy proved to be effective for the synthesis of ICE inhibitors, maintaining key hydrogen bond interactions with the enzyme and invoking a preferred conformation for binding. Several compounds exhibited IC(50) values less than 10nM in a caspase-1 enzyme assay and less than 100nM in a THP-1 whole cell assay measuring IL-1beta production. Two compounds, 13c and 13j, were found to have good oral bioavailability (>50%) in rats when administered as prodrugs. PMID- 17127069 TI - DAT/SERT selectivity of flexible GBR 12909 analogs modeled using 3D-QSAR methods. AB - The dopamine reuptake inhibitor GBR 12909 (1-{2-[bis(4 fluorophenyl)methoxy]ethyl}-4-(3-phenylpropyl)piperazine, 1) and its analogs have been developed as tools to test the hypothesis that selective dopamine transporter (DAT) inhibitors will be useful therapeutics for cocaine addiction. This 3D-QSAR study focuses on the effect of substitutions in the phenylpropyl region of 1. CoMFA and CoMSIA techniques were used to determine a predictive and stable model for the DAT/serotonin transporter (SERT) selectivity (represented by pK(i) (DAT/SERT)) of a set of flexible analogs of 1, most of which have eight rotatable bonds. In the absence of a rigid analog to use as a 3D-QSAR template, six conformational families of analogs were constructed from six pairs of piperazine and piperidine template conformers identified by hierarchical clustering as representative molecular conformations. Three models stable to y value scrambling were identified after a comprehensive CoMFA and CoMSIA survey with Region Focusing. Test set correlation validation led to an acceptable model, with q(2)=0.508, standard error of prediction=0.601, two components, r(2)=0.685, standard error of estimate=0.481, F value=39, percent steric contribution=65, and percent electrostatic contribution=35. A CoMFA contour map identified areas of the molecule that affect pK(i) (DAT/SERT). This work outlines a protocol for deriving a stable and predictive model of the biological activity of a set of very flexible molecules. PMID- 17127071 TI - Ultrastructural types of alveolar macrophages in bronchoalveolar lavages from patients with pulmonary sarcoidosis. AB - By routine applied quantitative BAL methods are particularly helpful for the diagnosis of pulmonary sarcoidosis. Here the morphology of the alveolar cells does not play a role. However, morphological and especially electron microscopic investigations might contribute to the clarification of the aetiology of this disease. In a prospective study we investigated the bronchoalveolar lavages (BALs) from 10 patients with recently histologically diagnosed, untreated pulmonary sarcoidosis. Commonly applied cytological and immunological BAL diagnostic techniques were accompanied by morphological investigations of alveolar cells, especially alveolar macrophages, using light and electron microscopy. All patients showed lymphocytic alveolitis with an increased number of CD4 positive lymphocytes as well as an increased CD4/CD8 ratio. A striking light microscopic finding was the great morphological variety of the alveolar macrophages. Electron microscopy revealed typical lymphocytes, neutrophils, and eosinophils as well as three different types of alveolar macrophages in all 10 patients: type I (approx. 30%) with a normal macrophage morphology, a vacuole rich type II (approx. 30%) with myelin-like structures and type III (approx. 40%) with electron-dense inclusions. The occurrence of intracellular myelin figures in type II macrophages is a hint for increased phagocytotic processes of surfactant with or without its overproduction in the sense of a secondary alveolar proteinosis. Numerous electron-dense inclusions in type III also indicate an increased macrophage activity that leads to an increased release of cytokines, which in turn can trigger an inflammatory reaction. PMID- 17127072 TI - A note on the half-life of 209Po. AB - The widely adopted value of (102+/-5)a for the (209)Po half-life, which is based on a single determination reported in 1956, appears to be in error by a large factor. Decay data from two separate primary standardizations of a (209)Po solution standard, conducted approximately 12 years apart, are inconsistent with the adopted value and its assigned uncertainty. An estimated half-life, larger than the adopted value by about 25%, is more consistent with the standardization data. A longer half-life is also supported by measurements on a recently standardized (210)Pb solution standard. PMID- 17127073 TI - Apico-aortic conduit. PMID- 17127074 TI - Poly(ADPR)polymerase-1 signalling of the DNA damage induced by DNA topoisomerase I poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines. AB - Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to DNA topoisomerase I (TOPO I) inhibitor-mediated apoptosis. We investigated the effects of combined treatments with the DNA topoisomerase I inhibitor Topotecan and the poly(ADPR)polymerase-1 inhibitor NU1025 in D54(p53wt) and U251(p53mut) glioblastoma cell lines. Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM NU1025 and a G2/M block of the cell cycle. We also evaluated, the influence of TPT+/-NU1025 treatment on PARP-1 and p53 activity. We got evidences of a TPT-dependent increase of PARP-1 auto modification level in both the cells. Moreover, in the D54(p53wt) cells we found that in co-treatments NU1025 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the p21 nuclear amount. Conversely, in U251(p53mut) cells we found that NU1025 incremented the TPT-dependent apoptosis characterised by PARP-1 proteolysis. Our findings suggest that the modulation of PARP-1 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status. PMID- 17127075 TI - Long-term inhibition of Rho kinase with fasudil attenuates high flow induced pulmonary artery remodeling in rats. AB - Accumulating evidences have demonstrated that RhoA/Rho-kinase pathway plays a pivotal role in various cellular functions. The aim of this study was to explore whether RhoA/Rho-kinase pathway is involved in the pathogenesis of high flow induced pulmonary hypertension and whether long-term inhibition of RhoA/Rho kinase pathway with fasudil could attenuate high flow induced pulmonary artery remodeling in rats. Wistar rats in the shunt groups and treated groups were underwent left common carotid artery-external jugular vein shunt operation, rats in control groups were sham-operated animals. Rats in treated groups received fasudil treatment, the others received same dose of saline. At week 4, 8 of the study, rats were underwent haemodynamics measurements, pulmonary artery morphologic assessments, detection of pulmonary artery smooth muscle cells (SMCs) proliferation and apoptosis. RhoA and Rho-kinase activity in pulmonary arteries were also analyzed. Compared with the control groups, exposure to high blood flow induced a significant elevation of right ventricle systolic pressure at week 8, significant increase of the mean percentage of media wall thickness (%MT) in moderate size pulmonary arteries both at weeks 4 and 8, marked elevation of right ventricle (RV) to left ventricle plus septum (LV+SP) weight ratio at week 8, significant increase of PCNA-positive SMCs percentage at week 4 and significant decrease of TUNEL-positive SMCs percentage both at weeks 4 and 8. High pulmonary blood flow also induced 3.19+/-0.28-fold increase of RhoA and 3.63+/-0.52-fold increase of Rho-kinase over the control group at week 4, 1.57+/-0.35-fold increase of RhoA and 2.36+/-0.39-fold increase of Rho-kinase over the control group at week 8. Compared with the shunt groups, fasudil treatment significantly suppressed Rho-kinase activity at both weeks 4 and 8, improved pulmonary hypertension at week 8, attenuated right ventricular hypertrophy at week 8, and enhanced pulmonary vascular remodeling both at weeks 4 and 8, which were associated with suppressed pulmonary artery smooth muscle cells proliferation at week 4 and apoptosis both at weeks 4 and 8. These results indicated that RhoA/Rho kinase mediated pathway participated in the process of high flow induced pulmonary artery remodeling; inhibition of Rho kinase with fasudil could attenuate pulmonary artery remodeling. PMID- 17127076 TI - Co-chaperone CHIP promotes aggregation of ataxin-1. AB - Recent studies demonstrated that co-chaperone/E3 ligase CHIP (C-terminus of hsp70 interacting protein) mediates the ubiquitylation and suppresses the aggregation of polyglutamine (polyQ) proteins, such as huntingtin or ataxin-3. In this study, we investigated the effects of CHIP on the degradation of another polyQ protein ataxin-1. Interestingly CHIP associates not only with the polyQ-expanded ataxin-1 but also with the normal ataxin-1. Moreover, by enhancing ataxin-1 ubiquitylation, CHIP over-expression leads to a reduction in the solubility of ataxin-1 and thus increases the aggregate formation, especially that of polyQ expanded ataxin-1. Domain analysis revealed that the TPR domain is required for the promotion of aggregation. By contrast, other co-chaperones or E3 ligases, such as BAG-1 or parkin, did not show similar effects on the aggregation of ataxin-1. Importantly, the effect of CHIP is impaired by the mutation of Ser776 of ataxin-1 whose phosphorylation is crucial for ataxin-1 aggregation. Our findings suggest that the role of CHIP in aggregation of polyQ proteins greatly varies depending on the context of full-length polyQ proteins. PMID- 17127077 TI - Molecular cloning, expression and purification of L-amino acid oxidase from the Malayan pit viper Calloselasma rhodostoma. AB - A cDNA encoding LAAO from the Malayan pit viper (Calloselasma rhodostoma) was cloned into an expression vector of the methylotropic yeast Pichia pastoris. The LAAO open reading frame was inserted after the alpha-MF-signal sequence. Upon induction soluble and active LAAO is produced and exported into the culture supernatant at a concentration of up to 0.4 mg/L. Recombinant LAAO was purified from this by ion exchange and molecular sieve chromatography to yield apparently homogeneous protein in quantities of approximately 0.25 mg/L growth medium. Expressed LAAO exhibits the same electrophoretic mobility as native LAAO (62 kDa) and exhibits approximately the same extent of glycosylation as authentic LAAO from snake venom. Catalytic properties and substrate specificity of recombinant LAAO are similar to those of native enzyme. PMID- 17127078 TI - Reliability and performance-dependent variations of muscle function variables during isometric knee extension. AB - Despite the common use of standardised methods analysing neuromuscular function during knee extension, there is a lack of test-retest reliability studies. Furthermore, for most of the investigated variables it is unknown which changes of values indicate an enhancement of performance. The aim of the present study was to investigate performance-dependent variations of muscle functions during isometric contraction of knee extensors and to examine test-retest reliability of their measurement methods. For test-retest reliability sports students completed three test sessions. Highly skilled athletes, sports students and untrained subjects were investigated to determine the performance-dependent variations. The following variables were analysed: maximal voluntary contraction (MVC), voluntary activation (VA), absolute muscle reaction time (AR), muscle endurance (ME), and EMG frequency analysis (MF) of m. vastus lateralis (VL), m. vastus medialis (VM) and m. rectus femoris (RF). RESULTS TEST-RETEST-RELIABILITY: A high reliability between session 1 vs. 2 and session 2 vs. 3 was shown for MVC (ICC=0.92 and .97), VA (0.92/0.95) and ME (0.87/0.95). ICC in AR (0.23) was low between the first and second session and moderate between the second and third session (0.74). MF of VL, VM and RF showed low ICC between sessions. PERFORMANCE DEPENDENT VARIATIONS: Significant differences in nearly all variables (except VA) were found between trained (athletes and sports students) and untrained subjects. PMID- 17127079 TI - Deformation-based morphometry of brain changes in alcohol dependence and abstinence. AB - Brain atrophy associated with chronic alcohol consumption is partially reversible after cessation of drinking. Recovering alcoholics (RA, 45+/-8 years) were studied with MRI within 1 week of entering treatment, with follow-up at 8 months. Light drinkers (LD) were studied with MRI twice 1 year apart. For each participant, deformation maps of baseline structure and longitudinal size changes between baseline and follow-up scans were created using nonlinear registration techniques. ANCOVA assessed group differences and regression methods examined relationships between deformation maps and measures of drinking severity or baseline atrophy. At baseline, RA showed significant atrophy in the frontal and temporal lobes. Longitudinally, abstainers recovered tissue volumes significantly faster than LD in parietal and frontal lobes. When comparing abstainers to relapsers, additional regions with significantly greater recovery in abstainers were temporal lobes, thalamus, brainstem, cerebellum, corpus callosum, anterior cingulate, insula, and subcortical white matter. Gray matter volume at baseline predicted volume recovery during abstinence better than white matter. Drinking severity was not significantly related to brain structural changes assessed with this method. Longitudinally, deformation-based morphometry confirmed tissue recovery in RAs who maintain long-term sobriety. Abstinence-associated tissue volume gains are significant in focal parts of the fronto-ponto-cerebellar circuit that is adversely affected by heavy drinking. PMID- 17127080 TI - Molecular phylogenetics of Caenogastropoda (Gastropoda: Mollusca). AB - Caenogastropoda is the dominant group of marine gastropods in terms of species numbers, diversity of habit and habitat and ecological importance. This paper reports the first comprehensive multi-gene phylogenetic study of the group. Data were collected from up to six genes comprising parts of 18S rRNA, 28S rRNA (five segments), 12S rRNA, cytochrome c oxidase subunit I, histone H3 and elongation factor 1alpha. The alignment has a combined length of 3995 base positions for 36 taxa, comprising 29 Caenogastropoda representing all of its major lineages and seven outgroups. Maximum parsimony, maximum likelihood and Bayesian analyses were conducted. The results generally support monophyly of Caenogastropoda and Hypsogastropoda (Caenogastropoda excepting Architaenioglossa, Cerithioidea and Campanilioidea). Within Hypsogastropoda, maximum likelihood and Bayesian analyses identified a near basal clade of nine or 10 families lacking an anterior inhalant siphon, and Cerithiopsidae s.l. (representing Triphoroidea), where the siphon is probably derived independently from other Hypsogastropoda. The asiphonate family Eatoniellidae was usually included in the clade but was removed in one Bayesian analysis. Of the two other studied families lacking a siphon, the limpet-shaped Calyptraeidae was associated with this group in some analyses, but the tent shaped Xenophoridae was generally associated with the siphonate Strombidae. The other studied hypsogastropods with an anterior inhalant siphon include nine families, six of which are Neogastropoda, the only traditional caenogastropod group above the superfamily-level with strong morphological support. The hypotheses that Neogastropoda are monophyletic and that the group occupies a derived position within Hypsogastropoda are both contradicted, but weakly, by the molecular analyses. Despite the addition of large amounts of new molecular data, many caenogastropod lineages remain poorly resolved or unresolved in the present analyses, possibly due to a rapid radiation of the Hypsogastropoda following the Permian-Triassic extinction during the early Mesozoic. PMID- 17127081 TI - Gobius lagocephalus: the world's most widespread nomen dubium. PMID- 17127082 TI - Unreliability of depopulated bovine ureteric xenograft for infra inguinal bypass surgery: mid-term results from two vascular centres. AB - INTRODUCTION: We report a two centre experience with a depopulated ureteric xenograft (SGVG 100), CryoLife Inc., GA, USA) for femoropopliteal revascularization in 12 patients with chronic critical limb ischemia. REPORT: Between 7 days and 18 months after implantation, 10 of 12 patients (1 lost to follow-up) had the graft explanted due to aneurysmal enlargement. At 5 years, only one graft was still patent and showed moderate signs of enlargement. CONCLUSION: The SGVG 100 is not a safe conduit for femoropopliteal bypass surgery. PMID- 17127083 TI - A painless method of ultrasonically assisted debridement of chronic leg ulcers: a pilot study. AB - OBJECTIVES: Devitalized tissue in a recalcitrant leg ulcer is common and may impede healing. The aim of this study was to evaluate the use of a non-invasive low frequency ultrasound device to debride chronic leg ulcers as an adjunct to compression bandages therapy. METHODS: 19 patients with leg ulceration of at least 6 months were recruited. Low frequency ultrasound at 25kHz was delivered by a portable Sonaca--180 via a handheld probe, using normal saline as the irrigation/coupling medium. The ultrasound was applied for 10-20 seconds per probe head area onto the ulcer. Each leg underwent treatment at an interval of 2 3 weeks with compression bandages reapplied at the end of the treatment. Serial colour photographs were taken to evaluate the response at each visit. RESULTS: Each patient received on average 5.7 treatments each ranged from 5-20 minutes depending on the ulcer size. Symptomatic relief (pain and odour reduction) was achieved in 6 patients. 7 patients achieved complete ulcer healing (mean ulcer size=4.72+/-SD 1.872cm(2)) but no response was observed in 8 patients. There were no major complications of the treatment which was relatively painless. CONCLUSIONS: The application of low frequency ultrasound debridement may heal some recalcitrant ulcers when standard compression regimens have failed. It is cheap and does not require admission. The role of simple wound cleansing requires further investigation. PMID- 17127084 TI - Laparoscopic vascular surgery: a systematic review. AB - OBJECTIVE: The objective of this systematic review is to evaluate the results of clinical studies on laparoscopic surgery for aorto-iliac disease. METHODS: A systematic review of the literature from 1966 to September 2006 on laparoscopic and robotic vascular surgery was performed. Only patient series containing more than 5 cases were included. Operative, clamping and anastomosis times, conversion, mortality and morbidity and hospital stay were evaluated. RESULTS: Thirty studies were identified. These were all descriptive and included 9 comparative studies. Operative times varied widely, the shortest being for hand assisted procedures (2.5-4 hours) and the longest for totally laparoscopic procedures (4-6.5 hours). Clamping times were all<1 hour in hand-assisted procedures while in other techniques clamping times from 1-2.5 hours were seen. The conversion rate varied from <5% up to 16% in smaller series. The mortality rate was approximately 5% and frequently caused by cardiac ischemia. A variety of problems ranging from minor local wound problems to cardiopulmonary- and renal insufficiency, bleeding, ureter lesions and graft thrombosis were described. Mean hospital stay for nearly all procedures was <1 week. CONCLUSIONS: Experience of laparoscopic surgery for aorto-iliac disease is still limited. Most study results are biased by patient selection. Only a few surgeons have mastered the required surgical technique and more data are needed to asses the clinical potential of this type of surgery, in comparison with the endovascular alternative. For wider implementation simplification of the surgical procedure seems necessary. PMID- 17127085 TI - Ontogeny of the Spitzenkorper in germlings of Neurospora crassa. AB - The Spitzenkorper (Spk) is a highly dynamic and pleomorphic complex located at the hyphal apex of filamentous fungi. Most studies revealing the structure and behavior of the Spk have been conducted on mature vegetative hyphae of filamentous fungi, including both main leading hyphae and branches. However, these reports do not address whether the observations can be extended to germ tubes. By enhanced phase-contrast video-microscopy and laser scanning confocal microscopy we have analyzed the intracellular changes prior to the appearance of a Spk in germlings of Neurospora crassa. Observations began at the early stages of spore germination and were carried out until a conspicuous Spk could be observed at the apex of germ tubes. Before a Spk could be observed, young germ tubes (<150 microm) displayed a uniform distribution of organelles such as nuclei, mitochondria, and cytoplasmic granules along the length of the cells. Once the germlings started reaching lengths of more than approximately 150 microm, visible organelles experienced a displacement towards the subapical region of the cell and a small exclusion zone free of organelles (0.6+/-0.3 microm) formed at the apex. The position of this exclusion zone within the apex seemed to determine the germling growth direction, which was highly erratic. Few minutes after it first appeared, upon growth of the germling, the exclusion zone started to become occupied by an accumulation of material that gradually concentrated into a light gray body that we describe as an immature Spk. During this phase the presence of a Spk in the apical dome was not constant. Approximately 30 min later, the immature Spk became more robust and gradually acquired its typical phase-dark appearance, while the growth direction of the germ tube became less wavering. The formation of a mature phase-dark Spk coincided with the stabilization of the growth direction of the germling, therefore suggesting that it is at this stage when the transition from germling to vegetative hypha occurs. PMID- 17127086 TI - Fitting of deuterium quadrupole echo spectra with multiple motional models. AB - Dynamic information is generally extracted from deuterium quadrupole echo spectra by matching a spectrum calculated for a particular motional model to the experimental spectrum. In this work, a set of computer programs has been written to facilitate fitting of calculated spectra to experimental spectra that represent from one to five motional models. The fitting program requires pre calculated libraries of spectra for the models of interest, and accomplishes the fitting either by a systematic method or by simulated annealing. The systematic method is convenient for fitting with one or two motional models, but the simulated annealing method is faster for two or more models, if the libraries are made up of hundreds of spectra. The parameter Q, with the standard deviation of the spectral points estimated as the standard deviation of the baseline noise, provides a stringent measure of goodness of fit. Acceptable fits of experimental data as judged by this criterion have not been found, even in the case of ring flip motion in phenylalanine-d(5) in which the fit may be judged acceptable by eye. An example of fitting with isotropic and methyl rotation motional models of alanine-d(3), which have distinct spectral patterns, shows that it is possible to obtain reasonably accurate estimates of the relative amounts of deuterium representing the different models, even from poorly fitted spectra. PMID- 17127087 TI - Pharmacokinetics of telithromycin using bronchoscopic microsampling after single and multiple oral doses. AB - OBJECTIVES: Bronchoscopic microsampling (BMS) is a new technique for repeated sampling of bronchial epithelial lining fluid (ELF) to obtain the pharmacokinetic profile of drugs. We analyzed the time versus concentration profiles of telithromycin in bronchial ELF obtained by BMS and compared these finding to those in plasma and alveolar ELF obtained by bronchoalveolar lavage (BAL). METHODS: Bronchial ELF samples were obtained from five healthy subjects using BMS probe at 0, 2, 3, 4, 6, 10 and 24h after single or multiple oral doses of 600 mg of telithromycin. Alveolar ELF was also obtained by BAL 3h after single or multiple oral doses of 600 mg of telithromycin. RESULTS: The areas under the concentration-time curve from 0 to 24h (AUC0-24) of telithromycin in plasma and bronchial ELF were 2.86+/-0.60 and 19.5+/-10.4 mg h/l after single treatment and 3.60+/-0.49 and 42.2+/-22.7 mg h/l after multiple treatments, respectively. Single and multiple oral doses of telithromycin produced significantly (p<0.05) higher AUC0-24 in bronchial ELF compared to those in plasma. While concentrations in bronchial ELF obtained by BMS were significantly lower than those in alveolar ELF obtained by BAL, they tended to be higher than those in plasma after multiple administration. The telithromycin concentrations obtained by BMS method were very consistent in bronchial ELF at different bronchi at one time point and at the same bronchus at different time points. CONCLUSIONS: Using the BMS technique, we could describe the pharmacokinetics of telithromycin in bronchial ELF. Furthermore, BMS was reasonably validated and reconfirmed to be a feasible and reliable method for measuring antimicrobial concentrations in bronchial ELF. PMID- 17127090 TI - [Forceps verses vacuum extraction: no, forceps AND vacuum extraction!]. PMID- 17127088 TI - Vasculitides induced by TNFalpha antagonists: a study in 39 patients in France. AB - TNFalpha antagonists are effective in the treatment of chronic inflammatory joint disease. Despite a good overall safety profile, they can induce a number of adverse effects, including autoimmunity and infections. A link between TNFalpha antagonists and vasculitides has been suggested. METHODS: Between December 2004 and January 2005, a nationwide survey was conducted among 1200 hospital-based rheumatologists and internists in France, who were asked to report cases of vasculitis in patients taking TNFalpha antagonists. RESULTS: The survey identified 39 cases (32 women) of vasculitis during TNFalpha antagonist therapy. The joint disease was rheumatoid arthritis (RA) in 34 patients (including four without rheumatoid factor), juvenile idiopathic arthritis in two patients, ankylosing spondylitis in two patients, and psoriatic arthritis in one patient. Mean disease duration was 14.1+/-8.7 years. The TNFalpha antagonist was etanercept in 21 patients, infliximab in 15, adalimumab in 2, and another drug in 1; mean treatment duration was 9.6 months. The manifestations of vasculitis involved the skin (n=32); peripheral nervous system (n=9); kidney (n=7); central nervous system (n=3); pleura (n=2), pericardium (n=2); and the lung, gallbladder, and heart (n=1 each). Antinuclear factor (ANF) was present in 22 patients, hypocomplementemia in 6, and antineutrophil cytoplasmic antibody in 5. Histology (30 biopsies from 27 patients) showed nonnecrotizing vasculitis in 12 patients, necrotizing vasculitis in 7, an inflammatory dermal infiltrate without vasculitis in 3, extravascular necrotic granulomas in 2, chilblain lupus in 1, and cicatricial fibro-inflammatory changes in 1. Renal biopsy in three patients showed extracapillary glomerulonephritis with IgA deposits (n=2) or active floccular necrosis against a background of glomerular sclerosis (n=1). TNFalpha antagonist therapy was stopped in 33 patients, among whom 18 recovered without further treatment and 14 required high-dose glucocorticoids and/or immunosuppressant therapy, which ensured symptom resolution within a few weeks. The remaining patient died with multiple organ failures. DISCUSSION: The relative contributions of TNFalpha antagonist therapy and of the underlying disease to the development of vasculitis cannot be determined. Features that suggest a causal link between TNFalpha antagonists and vasculitis include the short time from TNFalpha antagonist initiation to vasculitis onset; the favorable response to discontinuation of TNFalpha antagonist therapy; and the development of systemic vasculitis in patients with rheumatoid factor-negative RA, in adults with juvenile-onset arthritis, and in patients with spondyloarthropathies. PMID- 17127091 TI - Programming good relations--development of the arbuscular mycorrhizal symbiosis. AB - The majority of plants live in symbiotic associations with fungi or bacteria that improve their nutrition. Critical steps in a symbiosis are mutual recognition and subsequently the establishment of an intimate association, which involves the penetration of plant tissues and, in many cases, the invasion of individual host cells by the microbial symbiont. Recent advances revealed that in the arbuscular mycorrhizal symbiosis with soil fungi of the order Glomeromycota, plant-derived signals attract fungal hyphae and stimulate their growth. Upon physical attachment of the fungal symbiont to the root surface, an active plant developmental program prepares the epidermal cells for penetration by the fungus. Thus, plants actively help symbiotic fungi to colonize their roots rather than just tolerating them. PMID- 17127092 TI - The International Workshops on Genotoxicity Testing (IWGT): history and achievements. PMID- 17127093 TI - Relevance of apoptosis in influencing recovery of hibernating myocardium. AB - BACKGROUND: Hibernating myocardium (HM) is viable but dysfunctional myocardium which can recover following revascularization. Myocyte necrosis is virtually absent in HM; however, cellular loss may take place by apoptosis, although this is controversial. AIM: To assess the presence of apoptosis and its relevance in HM. METHODS: During coronary artery by-pass surgery (CABG), 21 patients underwent transmural biopsy in the dysfunctional left anterior descending artery tributary area of the left ventricle (LV), with kinetic recovery at follow-up, thus fulfilling the HM criteria. All patients underwent echocardiographic follow-up at 12 months. All biopsies were evaluated by light microscopy, electron microscopy (EM), and molecular analysis. RESULTS: All biopsies were structurally altered, showing increased fibrosis and myocytes with variable size. Myocyte dedifferentiation was not detected by immunohistochemistry or EM. On stepwise linear regression, 1 year LVEF was predicted by the apoptotic index (beta=-0.973, p=0.002), the normotrophic cell percentage (beta=0.449, p=0.038), and mean fibrosis (beta=-0.412, p=0.51). CONCLUSIONS: Our biopsy study detected a wide range of morphological substrate heterogeneity in HM with degenerative features. We have demonstrated for the first time in humans that myocyte apoptosis is an important phenomenon in HM, negatively influencing LV functional recovery after CABG. PMID- 17127094 TI - Endogenous inhibitors of hypertrophy in concentric versus eccentric hypertrophy. AB - Left ventricular (LV) hypertrophy (LVH) is an adaptive response to hemodynamic overload, but also contributes to the pathogenesis of heart failure. LVH can be concentric (cLVH) but subsequent dilatation and progression to eccentric hypertrophy (eLVH) may lead to global pump failure. Recently, several endogenous molecular inhibitors of hypertrophy have been identified. Using real-time PCR, we compared the myocardial mRNA expression of these inhibitors in pressure-overload induced cLVH (severe aortic stenosis) and in volume overload-induced eLVH (severe mitral regurgitation) in patients, and during the progression from cLVH to eLVH in pressure overload in rat. Each of these genes showed a unique temporal expression profile. Strikingly, except for SOCS-3, changes in gene expression of these negative regulators in rat cLVH and eLVH vs sham were recapitulated in human cLVH and eLVH. In particular, VDUP-1 and MCIP-1 were high in cLVH but expression levels were normal in eLVH, both in rat and human. These data indicate that during the progression of LVH, both in pressure and volume overload, expression levels of endogenous inhibitors of hypertrophy are modified and that these changes may have pathophysiological significance. In particular, MCIP-1 (the endogenous calcineurin inhibitor) and VDUP-1 (the endogenous inhibitor of thioredoxin) are potential molecular switches in the progression of LV hypertrophy. PMID- 17127095 TI - Effect of wearing a mouthguard on the vestibulocollic reflex. AB - It has been speculated that the use of a mouthguard improves athletic ability such as muscular strength and equilibrium. The postural system is equipped with response patterns that correct for unexpected perturbations. These responses are driven by immediate feedback from visual, vestibular, and somatosensory information. These are integrated by the central nervous system. We analysed the possibility that wearing a mouthguard influences vestibular information via the vestibulocollic reflex (VCR) in the sagittal plane. The input of the VCR is vestibular afferent activity and its output is neck muscle activation. With the subject in the supine position, the apparatus used in this study induces the VCR by subjecting the head to abrupt vertical acceleration by sudden free fall under the head's own weight. Surface electromyographic (EMG) recordings in the sternocleidomastoid muscles (SCM) of 14 participants were analysed. There were no significant differences in the amplitude of the VCR within the left and right SCM whether subjects were or were not wearing a mouthguard (P=0.3227 within left SCM; P=0.9686 within right SCM). These results suggest that wearing a mouthguard has no effect on the VCR in the sagittal plane when supine, and so that vestibular sensory information is unaffected by wearing a mouthguard, in this context. Further research is required to examine whether this also holds true in more functional, upright and dynamic body positions. PMID- 17127096 TI - Brain CHIP: removing the culprits in neurodegenerative disease. AB - A factor that is common to the most-frequent neurodegenerative diseases is the accumulation of abnormal proteins that are associated with cellular dysfunction. Contrary to years of speculation, recent evidence suggests that soluble intermediates--not the visible pathological aggregates associated with disease- are the cause of neurotoxicity. These findings suggest that aggregate formation might be an adaptive stress response that is facilitated by neuronal protein triage molecules. In particular, the molecular co-chaperone CHIP (C terminus of HSC70-interacting protein) has been linked to several of these disorders, serving as a crucial catalyst for the ubiquitination of several heat shock protein (HSP)70 client proteins that are involved in neurodegenerative disease. Therefore, understanding the mechanisms that are involved in CHIP-mediated protein trafficking might provide invaluable clues to neuronal function, both in normal and diseased conditions. PMID- 17127097 TI - 'Coumadin ridge' in the left atrium demonstrated on three dimensional transthoracic echocardiography. AB - An echogenic band like structure was seen in the left atrium on two dimensional transthoracic echocardiography (2D TTE). Full volume three dimensional (3D) TTE and colour Doppler established the surrounding anatomical landmarks, and demonstrated the absence of obstruction related to this band. 3D TTE confirmed that this band like structure was consistent with the ridge between the left atrial appendage and left superior pulmonary vein ('warfarin/coumadin ridge'). PMID- 17127098 TI - Laser Doppler flowmetry for assessment of myocardial microperfusion in the beating rat heart. AB - Although laser Doppler flowmetry (LDF) is widely used for measuring microperfusion, it is rarely used to measure coronary microcirculation. The present in vivo study investigated the use of LDF to measure myocardial microperfusion in beating rat hearts. Ascending aortic flow and other hemodynamic parameters were simultaneously recorded. A needle probe with a holder was adhered to the epicardium of the left ventricular myocardium close to the left anterior descending coronary artery in an anaesthetized open-chest rat. Myocardial microperfusion was measured in response to bolus intravenous administration of both two representative vasodilators (captopril and nifedipine) and a vasoconstrictor (pituitrin). Myocardial microperfusion was found to be predominately diastolic, and in an opposing phase to the ascending aortic flow. Captopril (5 or 10 mg/kg) increased the initial myocardial microperfusion phase. Nifedipine at 75 microg/kg caused a sustained myocardial microperfusion elevation with a peak increase of 7.1+/-1.1%, but this was not observed using 150 microg/kg nifedipine. Both drugs caused an increase in the cardiac index. In contrast, myocardial microperfusion decreased (28.7+/-0.1% maximum decrease) in response to 1 IU/kg pituitrin. In conclusion, LDF provided a means of assessing myocardial microperfusion in beating rat hearts, and can be applied to evaluate the coronary microcirculation response to drugs. PMID- 17127099 TI - Is there a role for reactive oxygen species in arterial medial elastocalcinosis? AB - Isolated systolic hypertension results from a gradual stiffening of large arteries, to which medial elastocalcinosis (calcification of elastic lamellae) contributes. There is compelling evidence that reactive oxygen species (ROS) are associated with several disease processes affecting the cardiovascular system, including hypertension. The present study was designed to investigate whether the inhibition of ROS production by alpha-lipoic acid can prevent vascular calcification. Sprague-Dawley rats were treated with warfarin (20 mg/kg/day) and vitamin K (15 mg/kg/day) (WVK) for 4 weeks to induce large artery calcification. Subgroups received either a normal diet or a diet supplemented with lipoic acid (1000 mg/kg/day). The WVK treatment produced a small elevation of aortic superoxide levels that did not reach statistical significance. Alpha-lipoic acid reduced the elevation below baseline levels. In rats treated with alpha-lipoic acid, the WVK-induced elevation of pulse wave velocity (an index of arterial stiffness), left ventricular hypertrophy, and aortic, femoral and carotid elastocalcinosis were not prevented. Although a contribution of oxidative stress has been suggested in the aging cardiovascular system, this alteration does not appear to contribute to the calcification process and the subsequent stiffening of large arteries in the animal model tested. PMID- 17127101 TI - Expression analysis of the family of 14-3-3 proteins in zebrafish development. AB - 14-3-3 proteins comprise a family of dimeric multi-functional proteins present in all eukaryotes, that are important in a whelm of ubiquitous biological processes. We have analyzed the genomic structure of all 14-3-3s from zebrafish comprising 11 genes and have analyzed their phylogeny. The gene family was cloned and its expression pattern in zebrafish embryogenesis was analyzed by whole mount in situ hybridization and microarray analysis with gene specific probes. We demonstrate that maternal mRNA of 14-3-3s is expressed evenly at the first cell division. At later stage all genes are expressed in a patterned way with, in most cases, intricate patterns in the developing brain. Our result shows distinct expression patterns of various genes. Microarray results show that differences in expression levels of highly similar 14-3-3 genes also occur in the adult stage. PMID- 17127100 TI - Comparison of the mechanisms underlying the relaxation induced by two nitric oxide donors: sodium nitroprusside and a new ruthenium complex. AB - We studied the mechanisms involved in the relaxation induced by nitric oxide (NO) donors, ruthenium complex ([Ru(terpy)(bdq)NO(+)](3+)-TERPY) and sodium nitroprusside (SNP) in denuded rat aorta. Both NO donors induced vascular relaxation independent of the agonist used in the pre-contraction. [Ru(terpy)(bdq)NO(+)](3+) and SNP activated guanylyl cyclase (GC) and K(+) channels. The production of cGMP induced by [Ru(terpy)(bdq)NO(+)](3+) - was higher than that obtained with SNP. The combination of GC inhibitor with K(+)channels blocker almost abolished the relaxation induced by the NO donors. The extracellular NO scavenger oxyhemoglobin reduced the potency without changing the maximum effect (Emax) of both NO donors. By using specific NO species scavengers, hydroxocobalamin and l-cysteine, we have identified the contribution of free radical NO (NO()) and nytroxil anion (NO(-)), respectively, to the rat aorta relaxation induced by both NO donors. The selective scavengers for NO() and NO(-) reduced the potency but not the Emax of [Ru(terpy)(bdq)NO(+)](3+). However, the NO(-) scavenger had no effect on the relaxation induced by SNP and NO() scavenger reduced only the potency to SNP. The inhibition of sarcoplasmic reticulum Ca(2+)-ATPase reduced only the potency of SNP without effect on the relaxation induced by [Ru(terpy)(bdq)NO(+)](3+). Our results demonstrate that both NO donors induce relaxation by activating the GC and K(+) channels. The NO() is the unique NO specie involved in the SNP-relaxation. On the other hand, the relaxant effect of [Ru(terpy)(bdq)NO(+)](3+) involves both NO() and NO(-), that produce higher concentration of cGMP. The inhibition of sarcoplasmic reticulum Ca(2+)-ATPase reduces the relaxation induced by SNP but it did not alter the relaxation induced by [Ru(terpy)(bdq)NO(+)](3+). PMID- 17127102 TI - The zebrafish orphan nuclear receptor genes nr2e1 and nr2e3 are expressed in developing eye and forebrain. AB - Mammalian Nr2e1 (Tailless, Mtll or Tlx) and Nr2e3 (photoreceptor-specific nuclear receptor, Pnr) are highly related orphan nuclear receptors, that are expressed in eye and forebrain-derived structures. In this study, we analyzed the developmental expression patterns of zebrafish nr2e1 and nr2e3. RT-PCR analysis showed that nr2e1 and nr2e3 are both expressed during embryonic and post embryonic development. To examine the spatial distribution of nr2e1 and nr2e3 during development whole-mount in situ hybridization was performed. At tailbud stage, initial nr2e1 expression was localized to the rostral brain rudiment anterior to pax2.1 and eng2 expression at the prospective midbrain-hindbrain boundary. During subsequent stages, nr2e1 became widely expressed in fore- and midbrain primordia, eye and olfactory placodes. At 24hpf, strong nr2e1 expression was detected in telencephalon, hypothalamus, dorsal thalamus, pretectum, midbrain tectum, and retina. At 2dpf, the initially widespread nr2e1 expression became more restricted to distinct regions within the fore- and midbrain and to the retinal ciliary margin, the germinal zone which gives rise to retina and presumptive iris. Expression of nr2e3 was exclusively found in the developing retina and epiphysis. In both structures, nr2e3 expression was found in photoreceptor cells. The developmental expression profile of zebrafish nr2e1 and nr2e3 is consistent with evolutionary conserved functions in eye and rostral brain structures. PMID- 17127103 TI - Bayesian network analysis of resistance pathways against HIV-1 protease inhibitors. AB - Interpretation of Human Immunodeficiency Virus 1 (HIV-1) genotypic drug resistance is still a major challenge in the follow-up of antiviral therapy in infected patients. Because of the high degree of HIV-1 natural variation, complex interactions and stochastic behaviour of evolution, the role of resistance mutations is in many cases not well understood. Using Bayesian network learning of HIV-1 sequence data from diverse subtypes (A, B, C, F and G), we could determine the specific role of many resistance mutations against the protease inhibitors (PIs) nelfinavir (NFV), indinavir (IDV), and saquinavir (SQV). Such networks visualize relationships between treatment, selection of resistance mutations and presence of polymorphisms in a graphical way. The analysis identified 30N, 88S, and 90M for nelfinavir, 90M for saquinavir, and 82A/T and 46I/L for indinavir as most probable major resistance mutations. Moreover we found striking similarities for the role of many mutations against all of these drugs. For example, for all three inhibitors, we found that the novel mutation 89I was minor and associated with mutations at positions 90 and 71. Bayesian network learning provides an autonomous method to gain insight in the role of resistance mutations and the influence of HIV-1 natural variation. We successfully applied the method to three protease inhibitors. The analysis shows differences with current knowledge especially concerning resistance development in several non-B subtypes. PMID- 17127104 TI - Oxidative damage to purines in DNA: role of mammalian Ogg1. AB - DNA damage caused by reactive oxygen species is ubiquitous to all living organisms. More than 60 different base lesions have been identified, and the majority of these are removed via the base excision repair pathway. This pathway appears to represent a highly conserved and ancient mechanism of defence counteracting spontaneous DNA decay. In this review, we describe in more detail the Ogg1 enzyme and its conserved action of removing the oxidised base, 7,8 dihydro-8-oxoguanine (oxo(8)G). Recent updates include the cancer-prone ogg1/myh double knockout mouse and an elegant study which looks at the ability of hOgg1 to distinguish between the mutagenic lesion, oxo(8)G, and the vast majority of normal bases. PMID- 17127107 TI - N1303K and IVS8-5T, clinical presentation within a family with atypical cystic fibrosis. AB - The CFTR genotype N1303K/IVS8-5T can cause very mild cystic fibrosis (CF) and congenital bilateral absence of the vas deferens (CBAVD). We report one family consisting of five affected patients in two generations, presenting minor symptoms of CF at different ages, segregating the CFTR mutations N1303K and IVS8 T5-TG13 in trans. Common features were chronic sinopulmonary symptoms and borderline or slightly elevated sweat chloride values. One patient had CBAVD. PMID- 17127106 TI - Down-regulation of DNA polymerase beta accompanies somatic hypermutation in human BL2 cell lines. AB - Somatic hypermutation (SHM) is a fundamental process in immunoglobulin gene maturation that results in increased affinity of antibodies toward antigens. In one hypothesis explaining SHM in human B cells, the process is initiated by enzymatic deamination of cytosine to uracil in the immunoglobulin gene V-region and this in turn triggers mutation-prone forms of uracil-DNA base excision repair (BER). Yet, an uncertainty with this model is that BER of uracil-DNA in mammalian cells is generally error-free, wherein DNA polymerase beta (pol beta) conducts gap-filling synthesis by insertion of bases according to Watson-Crick rules. To evaluate this inconsistency, we examined pol beta expression in various SHM proficient human BL2 cell line subclones. We report that expression of pol beta in SHM proficient cell lines was strongly down-regulated. In contrast, in other BL2 subclones, we found that SHM was deficient and that pol beta expression was much higher than in the SHM proficient subclones. We also found that overexpression of recombinant human pol beta in a SHM proficient subclone abrogated its capacity for SHM. These results suggest that down-regulation of the normal BER gap-filling DNA polymerase, pol beta, accompanies induced SHM in BL2 cells. This is consistent with the hypothesis that normal error-free BER must be silenced to make way for an error-prone BER process that may be required during somatic hypermutation. PMID- 17127108 TI - Exhaled nitric oxide in severe obesity: effect of weight loss. AB - Exhaled nitric oxide (NO) is a recognized biomarker in the lower respiratory tract. The effect of large variation in body mass on exhaled NO in the same individuals is not well known. The aim of the study was to evaluate both the effect of severe obesity and the influence of weight reduction on exhaled NO. A consecutive series of 24 uncomplicated obese patients (OB), who had laparoscopic adjustable gastric banding (LAGB) and 15 healthy controls (HC) were studied. Body mass index (BMI), exhaled NO and respiratory function tests were assessed. Exhaled NO was lower in obese in comparison to HC (12.0+/-3.6ppb versus 15.8+/ 4.0ppb, p=0.0035). A significant positive correlation was found between exhaled NO and BMI in HC, which was not evident in OB. Among the respiratory indexes, functional residual capacity was significantly associated to exhaled NO. After 1 year, 12 obese patients undergone to LAGB were re-evaluated. Mean BMI (kg/m(2)) decreased from 44.8 before surgery to 32.3 post-operatively. The exhaled NO increased from 11.8+/-3.2ppb before surgery to 14.9+/-3.1ppb 1 year post operatively (p=0.0023, n=12). In conclusion exhaled NO is consistently reduced in severe obesity and it is restored after weight reduction. The relationship between exhaled NO, large body mass excess and decrease of resting lung volume in severe obesity deserves further studies. PMID- 17127109 TI - Quantification of organic eluates from polymerized resin-based dental restorative materials by use of GC/MS. AB - Residual monomers, additives and degradation products from resin-based dental restorative materials eluted into the oral cavity may influence the biocompatibility of these materials. Emphasis has been placed on studies addressing cytotoxic, genotoxic and estrogenic potential of these substances. A prerequisite for analyzing the potential of exposure to eluted compounds from dental materials is reliable quantification methods, both real time and accelerated measurements. The purpose of the present study was to quantify nine eluates; 2-hydroxyethyl methacrylate (HEMA), hydroquinone monomethyl ether (MEHQ), camphorquinone (CQ), butylated hydroxytoluene (BHT), ethyl 4 (dimethylamino)benzoate (DMABEE), triethylene glycoldimethacrylate (TEGDMA), trimethylolpropane trimethacrylate (TMPTMA), oxybenzone (HMBP) and drometrizole (TIN P) leaching from specimens of four commonly used resin-based dental materials in ethanol and an aqueous solution. All analyses were performed by use of GC/MS, each component was quantified separately and the results presented in microg mm(-2). This study has shown that elution from various materials differs significantly, not only in the types of eluates, but also regarding amounts of total and of single components. A high amount of HMBP, a UV stabilizer with potential estrogenic activity, was detected from one material in both solutions. PMID- 17127110 TI - Development of an in vitro incubation procedure for screening of CYP2D6 intrinsic clearance. AB - The in vitro intrinsic clearances (CL(int)) for the metabolism of p methoxymethamphetamine (PMMA) and fluoxetine by the CYP2D6 enzyme were calculated using a steady-state (SS) approach and a new general enzyme (GE) method, which measures the formation of product and the depletion of substrate as a function of time. For PMMA, the SS experiment resulted in a CL(int) of 2.7+/-0.2 microL pmol 2D6(-1)min(-1) and the GE experiment resulted in a CL(int) of 3.0+/-0.6 microL pmol 2D6(-1)min(-1). For fluoxetine, the SS experiment resulted in a CL(int) of 0.33+/-0.17 microL pmol 2D6(-1)min(-1) and the GE experiment resulted in a CL(int) of 0.188+/-0.013 microL pmol 2D6(-1)min(-1). We used two kinetic modeling techniques that can accommodate atypical kinetic models. We also show that the addition of fluoxetine results in a 10-fold decrease in the observed intrinsic clearance of PMMA, confirming that fluoxetine is a potent inhibitor of the liver enzyme CYP2D6. PMID- 17127111 TI - High performance liquid chromatography-tandem mass spectrometry (HPLC/MS/MS) assay for chiral separation of lactic acid enantiomers in urine using a teicoplanin based stationary phase. AB - A novel method for the separation and simultaneous determination of urinary D- and L-lactic acid enantiomers by high performance liquid chromatography-tandem mass spectrometry (HPLC/MS/MS) is presented. The chiral separation was optimized on a Chirobiotic teicoplanin aglyocone (TAG) column. Most interestingly, the addition of water in small volume fraction to the polar organic mobile phase was found to significantly improve the chromatography. Calibration curves were linear (r2>0.9950) over the range 3-1000 mg/L for L-lactic acid and 0.5-160.8 mg/L for D lactic acid. The limit of detection (LOD) (S/N=3) and limit of quantification (LOQ) (S/N=10) were determined experimentally (n=3) to be 0.2 and 0.5mg/L for L lactic acid and 0.4 and 1.3 mg/L for D-lactic acid, respectively. The normal patient range of L-lactic acid was 1-20 microg/mg creatinine with an elevated value of 85 microg/mg creatinine. For D-lactic acid, the range of normal values were between 0 and 5 microg/mg creatinine with an elevated value of 40 microg/mg creatinine. Finally, the validated method allows for rapid analysis with a total run time of 7.5 min. PMID- 17127112 TI - On the accuracy of perceived parental height in a native Amazonian society. AB - Studies of secular trends in adult height in rural pre-literate societies are likely to show no change owing to random measurement error in age. In such societies, adults lack birth certificates and guess when estimating their age. We assess the accuracy of perceived height of the same-sex parent to estimate secular trends. We tested the method among the Tsimane', a native Amazonian society of farmers and foragers in Bolivia. Subjects included 268 women and 287 men >20 years of age. Over half the sample reported inaccurately the height of their same-sex living parent, with a tendency to report no difference when, in fact, differences existed. Results highlight the pitfalls of using perceived parental height to examine secular trends in adult height among the Tsimane', though the method might yield accurate information in other societies. We discuss possible reasons for the low accuracy of Tsimane' estimates. PMID- 17127113 TI - Fluorine-substituted hydroxyapatite scaffolds hydrothermally grown from aragonitic cuttlefish bones. AB - Porous hydroxyapatite scaffolds with different levels of fluorine substitution (46% and 85%) on the OH sites were produced via hydrothermal transformation of aragonitic cuttlefish bones at 200 degrees C and calcination at temperatures up to 1200 degrees C. The increasing level of F substitution reduces the kinetics and probably the yield of the reaction. The incorporation of F in the lattice of hydroxypatite caused a lowering of the unit cell volume due to reduction of the length of the a-axis. The crystallites formed were close in size to bone-like apatite and were orientated along the a-axis rather than the c-axis. There was evidence of AB-type carbonated apatite. PMID- 17127114 TI - Rumour of angels and heavenly midwives: anthropology of transpersonal events and childbirth. AB - Some contemporary women can experience non-ordinary states of consciousness when childbearing. The purpose of this paper is to bring a 'transpersonal' frame to these non-ordinary states of consciousness (hereafter: NOSC). Transpersonal psychology is an interdisciplinary movement in Western science that studies 'religious', 'peak' or 'healing' experiences in different cultures and social contexts. Between 2001 and 2006 in Auckland, New Zealand, while engaged in anthropological fieldwork, I collected stories from mothers, fathers, and midwives who had participated in transpersonal events during childbirth. I will compare the local women's NOSC with ethnographic accounts of spirit-possession and its relationship to indigenous midwifery then revisit and reconstruct the witch-hunts of Medieval Europe from this perspective. Midwives are encouraged to learn to identify and support women's NOSC during labour and birth as many women find strength and wisdom by passing through these states in labour. The subject is also critical to men, whether they are present with women and birth as fathers or health professionals. The hoped for result of this inquiry is to revalorise NOSC among birth-giving mothers, and to educate birth attendants in this field. PMID- 17127115 TI - Enhancing the midwife-woman relationship through shared decision making and clinical guidelines. AB - The active and passive voice allocated to women within maternity service guidelines helps construct the nature of decision making. The aim of this paper is to demonstrate that by allocating women an active voice within practice guidelines serves the interest of all parties within the health care relationship. Clinical guidelines were reviewed, and electronic databases and text were searched. The findings of this paper support that applying the principles of a shared decision making framework, within clinical practice guidelines, can assist the development of a partnership relationship between midwives and women. PMID- 17127116 TI - An in vitro study of ultrasound signal loss across simple fractures in cortical bone mimics and bovine cortical bone samples. AB - Measurements have been performed on Sawbones and bovine cortical bone samples at 200 kHz using an axial transmission technique to investigate the factors that determine how ultrasonic waves propagate across a simulated fracture. The peak amplitude of the first arrival signal (FAS) was studied. Results taken from intact specimens were compared with those produced when a simple transverse fracture was introduced. These fracture simulation experiments were found to be consistent with Finite Difference modelling of the experimental conditions. The peak amplitude showed a characteristic variation across the fracture caused by interference between reradiated and scattered/diffracted waves at the fracture site and a net Fracture Transmission Loss (FTL). For small fracture gaps, the change in amplitude was sensitive to the presence of the fracture. This sensitivity suggests that this parameter could be a good quantitative indicator for the fracture healing process assuming the relative change in this parameter brought about by healing is measurable. PMID- 17127117 TI - Osteogenesis imperfecta type VI in childhood and adolescence: effects of cyclical intravenous pamidronate treatment. AB - Cyclical intravenous treatment with pamidronate is of clinical benefit in children with moderate to severe osteogenesis imperfecta (OI) types I, III and IV, but there is no information on the effects of this treatment on the newly described OI type VI. Here, we report on the results of 3 years of pamidronate treatment in 10 children and adolescents with OI type VI (age range 0.8 to 14.5 years, three girls). Treatment effects were compared to those of 10 patients with OI types I, III, and IV, who were matched for age and disease severity (based on height and lumbar spine areal bone mineral density). During pamidronate therapy, lumbar spine areal bone mineral density z scores increased and lumbar spine vertebral bodies improved in shape. Iliac bone histomorphometry showed a tendency to higher cortical thickness (+53%, P=0.06) but the mineralization defect, a characteristic feature of OI type VI, did not change during pamidronate treatment. Annualized fracture incidence decreased from 3.1 per year before treatment to 1.4 fractures per year during treatment (P<0.05). Regarding mobility, the Pediatric Evaluation of Disability Inventory gross motor score increased by 42% during pamidronate treatment (P<0.005). Significant improvements were also found for age-related z scores of maximal isometric grip force. In comparison to the OI control group, the fracture incidence was higher and the gross motor scores were lower in OI type VI, both before and after pamidronate treatment (P<0.05 for each parameter). No differences were found between the groups for changes in densitometric measures and cortical thickness during pamidronate treatment. Our results suggest that 3 years of intravenous pamidronate therapy led to improvements in bone mineral mass, gross motor function, muscle force and fracture incidence in patients with OI type VI. However, the gains in mobility scores and reductions in fracture incidence during pamidronate treatment are less than in other OI types. PMID- 17127120 TI - Advancing injury prevention and trauma care in North America and globally. AB - Injury is a major global health problem. This article reviews ways in which the toll from injury can be lowered through the spectrum of injury control, including surveillance, prevention, and trauma care. There is room for improvement in the application of scientifically based, proved interventions at all points in the spectrum in all countries. The greatest attention is needed in low- and middle income countries, however, where most of the world's people live, where injury rates are higher, and where few injury control activities have yet been undertaken. PMID- 17127121 TI - Trauma systems. AB - The major goal of a trauma system is to enhance the community health. This occurs through a process of assessment, policy development, and ongoing assurance. This can be achieved by (1) identifying risk factors in the community and creating solutions to decrease the incidence of injury, (2) providing optimal care during the acute and the late phase of injury, including rehabilitation, and (3) maintaining the objective to decrease overall injury-related morbidity and mortality and years of life lost. Disaster preparedness also is an important function of trauma systems, and using an established trauma system network facilitates the care of victims of natural disasters or terrorist attacks. PMID- 17127122 TI - Prehospital care of the injured: what's new. AB - Emergency medical services (EMS) play a critical role in the trauma system as the point of initial patient care and stabilization and in determining the regional flow of patients and the commitment of resources to the critically injured. Trauma surgeons and emergency physicians need to be involved in the organizational planning of EMS systems to ensure that uniform patient care protocols are developed for triage and treatment. Ongoing efforts should focus on addressing national variability in care provided after injury to ensure optimal outcome for patients in all regions. Through additional research, the best practice and optimal EMS system design will continue to be defined. PMID- 17127123 TI - New developments in fluid resuscitation. AB - Hemorrhagic shock is the leading cause of death in civilian and military trauma. Effective hemorrhage control and optimal resuscitation are the main goals in the management of severely injured patients. This article addresses the changing trends in fluid resuscitation in regards to who, when, and how. Much of these changing trends are caused by the recognition that the current method of resuscitation with crystalloid fluids may not be optimal and may even have detrimental consequences. This article summarizes a number of studies that have evaluated the cellular toxicities of commonly used resuscitation fluids, to highlight the need for the development of new fluids. PMID- 17127124 TI - Damage control in trauma: laparotomy wound management acute to chronic. AB - Damage control surgery is fundamental to operative trauma care. Prophylactic application of open abdomen techniques has led to avoidance of a great deal of the organ dysfunction associated with abdominal compartment syndrome. Surgeons are learning about management of large open abdominal wounds. There seems to be a general consensus regarding acute management of these wounds. Institutions are using staged techniques of management. Getting open wounds closed as soon as possible leads to fewer complications. The acute use of vacuum wound may provide for early secondary closure. There is less study focused on optimal definitive reconstructive techniques. Further study in all of these areas will lead to improved outcomes. PMID- 17127125 TI - Thoracic trauma: when and how to intervene. AB - Trauma is the leading cause of death in patients younger than 40 years of age. Thoracic injuries are common and often can be managed by tube thoracostomy. In many patients, however, the thoracic injuries must be repaired surgically in one of three time periods: immediate, urgent, or delayed thoracotomy. In this article, we describe the general approach to effectively managing thoracic trauma patients. We review common injuries and scenarios that may be encountered by the surgeon and discuss the considerations and variables that enter into the decision making process for operative intervention. PMID- 17127126 TI - Care of central nervous system injuries. AB - The primary method of improving outcome from traumatic brain injury is through avoiding secondary insults to the injured brain. Although surgery is important, most management is critical care. Evidence-based guidelines continue to be developed to assist in directing care. With modern monitoring systems, a physiologic-based approach is increasingly applicable, allowing focused treatment for intracranial hypertension and ischemia. It is important to balance and integrate the care of the injured brain into the overall care of the polytrauma patient. PMID- 17127127 TI - Lessons learned from modern military surgery. AB - The era of global terrorism and asymmetric warfare heralded by the September 11, 2001 attacks on the United States have blurred the traditional lines between civilian and military trauma. The lessons learned by physicians in the theaters of war, particularly regarding the response to mass casualties, blast and fragmentation injuries, and resuscitation of casualties in austere environments, likely resonate strongly with civilian trauma surgeons in the current era. The evolution of a streamlined trauma system in the theaters of operations, the introduction of an in-theater institution review board process, and dedicated personnel to collect combat casualty data have resulted in improved data capture and realtime, on-the-scene research. PMID- 17127128 TI - Thermal and electrical injuries. AB - Through progress in wound management, resuscitation, intensive care treatment, and a coordinated rehabilitation process, modern burn care has been able to deliver substantial increases in survival and improvement in functional outcomes for burn victims. The development of regionalized burn centers has contributed greatly to this progress. As the field of burns matures, burn centers are preparing to meet future challenges through collaborative efforts in disaster management and outcomes research. PMID- 17127129 TI - Pediatric injuries: prevention to resolution. AB - Despite improved education and prevention initiatives, trauma remains the leading cause of death in children. A variety of preventative measures have been developed to decrease the morbidity and mortality, and the financial burden on the health care system. This article discusses injury prevention strategies, issues in prehospital care, and key points of initial resuscitation. In addition, the major injury patterns are described with attention paid to the diagnosis and management of patients with multiple traumatic injuries. PMID- 17127130 TI - Injury in the elderly and end-of-life decisions. AB - The elderly constitute the fastest growing sector of the population of the United Stated and geriatric trauma patients are presenting for care with increasing frequency. These patients are challenging particularly because of their vulnerability to severe injury, limited physiologic response to stress, and frequent presence of comorbid medical conditions complicating care. Many elderly trauma victims require prolonged intensive care and some fail to improve or succumb despite the best efforts because of the extent of their injuries and their underlying disease. These patients may present profound ethical challenges for trauma surgeons as the goals of care shift from salvage to end-of-life care. PMID- 17127131 TI - Environmental cold-induced injury. AB - More than 650 deaths from hypothermia occur each year in the United States. Even minor deviation from normal temperature leads to important symptoms and disability. The most significant risk factors are advanced age, mental impairment, substance abuse, and injury. This article examines the incidence of hypothermia, its detrimental effect on trauma patients, and methods of rewarming the hypothermic patient. It also looks at the controversial protective role hypothermia might play in shock, organ transplantation, cardiac arrest, and brain injury. Finally, it examines cold injuries, including frostbite, chilblain, and trench foot, and makes recommendations for their treatment. PMID- 17127132 TI - SR147778, a CB1 cannabinoid receptor antagonist, suppresses ethanol preference in chronically alcoholized Wistar rats. AB - This study investigated the effect of the new CB1 cannabinoid receptor antagonist, SR147778, on ethanol preference in chronically alcoholized Wistar rats. In study 1, SR147778, at doses of 0.3, 1, or 10 mg/kg/day (mg/kg/d) intraperitonealy (ip), was administered during chronic pulmonary ethanol intoxication for 30 days. The rats were then exposed to a two-bottle choice (ethanol 10% v/v vs. water) for at least 30 days. Neither 0.3 nor 1 mg/kg/d had any effect on ethanol preference. In contrast, the high dose induced a significant transient increase in ethanol intake between days 6 and 10. In study 2, SR147778, at doses of 0.3, 1, or 10 mg/kg/d ip, was administered during the free-choice period after chronic alcoholization. Both ethanol preference and intake were significantly reduced only for 1 and 10 mg/kg/d. These results reinforce the hypothesis that the cannabinoid CB1 receptor is part of the neural substrate mediating alcohol intake and the motivational properties of alcohol. When these results are compared with those obtained with SR141716 (Rimonabant) on ethanol preference, we observed that (1) coadministration of 10 mg/kg/d SR147778 during chronic alcoholization induced a shorter transient increase of ethanol intake than Rimonabant and (2) SR147778 treatment during the free-choice period at doses of 1 and 10 mg/kg/d decreased ethanol intake more dramatically than SR141716 which, furthermore, continued for the duration of the free choice. PMID- 17127133 TI - The effects of moderate neonatal ethanol exposure on eyeblink conditioning and deep cerebellar nuclei neuron numbers in the rat. AB - Heavy, bingelike patterns of exposure to ethanol during a portion of the early postnatal period in the rat, a time of rodent brain development corresponding to the human third trimester, has been shown to deplete cerebellar neurons and to produce deficits in cerebellar-dependent tasks. In the current study, we examined the impact of more moderate ethanol exposure, during an extended portion of the rat third trimester equivalent, on cerebellar-dependent learning (eyeblink conditioning) and deep cerebellar nuclei neuron numbers. Neonatal rats received 0, 1, 2, or 3g/kg/day of ethanol in milk formula via a single intragastric intubation each day across postnatal days 2-11, or were left untreated. Peak BACs for ethanol-exposed rats were 50, 150, and 225 mg/dl, respectively. Rats underwent eyeblink conditioning as young adults (70 days of age) and deep cerebellar nuclei neuron numbers were assessed at 100 days of age. In Experiment 1, all rats showed normal responsiveness to periorbital stimulation prior to conditioning. The 3-g/kg/day group was impaired in eyeblink conditioning and possessed fewer deep cerebellar nuclei neurons. A trend toward impairment was observed in the 2-g/kg/day group. However, the 0-g/kg/day group was also impaired in eyeblink conditioning. In Experiment 2, the unconditioned stimulus pretest phase was eliminated, the 0-g/kg/day group learned normally, and both the 2- and 3-g/kg/day groups were again impaired. These results suggest that more moderate doses of ethanol during the rat third-trimester equivalent can produce long-term effects on the cerebellum. PMID- 17127134 TI - Chronic intermittent ethanol exposure during adolescence reduces the effect of ethanol challenge on hippocampal allopregnanolone levels and Morris water maze task performance. AB - Chronic intermittent ethanol exposure (CIEE) in adolescent rats has been shown to produce long-lasting hypnotic, metabolic, and functional tolerance. Recently, it has been hypothesized that allopregnanolone mediates some effects of ethanol, including ethanol-induced impairments in the performance of the Morris Water Maze Task (MWMT). The current studies explore the relationship between cortical and hippocampal allopregnanolone levels and ethanol-induced impairments in the MWMT following CIEE treatment in adolescent rats. Adolescent rats were administered 5.0 g/kg ethanol or saline every 48 h for a 20-day period beginning on postnatal day (P) 30. Training in the spatial version of the MWMT occurred on nontreatment days. Following completion of CIEE treatment and training, MWMT performance was tested 30 min after ethanol (2.0 g/kg) or saline challenge on P 50 and P 62. A separate group of rats were CIEE treated and received an ethanol (2.0 g/kg) or saline challenge on P 50 or 62, and were used for hippocampal and cortical allopregnanolone determination. CIEE during adolescence produced tolerance to both ethanol-induced impairments in the MWMT and ethanol-induced allopregnanolone levels in the hippocampus on P 50. However, when animals were tested at P 62, the reduction in ethanol-induced MWMT impairments found in CIEE rats was reversed and allopregnanolone levels from both saline or ethanol challenge were increased above levels found in control animals. Taken together, these results suggest that CIEE during adolescence produces tolerance to ethanol-induced impairments in MWMT and corresponding changes in ethanol-induced allopregnanolone levels in the hippocampus. Furthermore, cognitive tolerance is reversible and time dependent, but the reversal of cognitive tolerance is not correlated with normalization of hippocampal allopregnanolone levels. PMID- 17127135 TI - A novel mouse model for the study of the inhibitory effects of chronic ethanol exposure on direct bone formation. AB - Excessive alcohol consumption has been reported to interfere with human bone homeostasis and repair in multiple ways. Previous studies have demonstrated that chronic ethanol exposure in the rat via an intragastric dietary delivery system inhibits direct bone formation during distraction osteogenesis (DO, limb lengthening). The opportunity to extend the rat ethanol studies to mice is now possible due to the development of mouse models of DO. This study employed a novel combination of liquid ethanol diet delivery and a murine DO model to test the hypothesis that chronic ethanol exposure would result in deficits in direct bone formation during DO in contrast to the pair-fed controls. Twenty-eight 12 month-old C57BL/6 male mice were acclimated to the Lieber-DeCarli liquid control diet #710027 (Dyets Inc.) over a 1-week period. The mice were separated into two diet groups (n=14/group): pair-fed control and ethanol (diet #710260). After being on diet for 82 days, all mice underwent placement of an external fixator and osteotomy on the left tibia. Following a 6-day latency period, distraction began at a rate of 0.075 mm twice a day (b.i.d.) for 14 days. The weight changes were equivalent for both groups. The hypothesis that chronic ethanol exposure would inhibit direct bone formation and produce skeletal toxicity was supported by radiographic (P=.011) and histologic (P=.002) analyses of the % new bone formation in the DO gaps, by peripheral quantitative computed tomography analysis of the total volumetric bone mineral density of the contralateral proximal tibias (P<.001) and contralateral femoral necks (P=.012), by three-point bending on the contralateral tibias (P<.001 energy to break), by pin site bone formation measures (P<.001), and by ethanol-associated increased adipocyte area (adjacent to the gap) percentages (P<.002). We conclude that this model can be used to study the mechanisms underlying inhibition of bone formation by chronic ethanol exposure and to test preclinical interventions. PMID- 17127136 TI - Chronic-alcohol exposure alters IGF1 signaling in H9c2 cells via changes in PKC delta. AB - Previously, we have demonstrated that chronic-alcohol exposure alters insulin like growth factor 1 (IGF1) signaling in adult rat heart cells. This report examines the effects of alcohol in vitro on the expression of protein kinase C (PKC) alpha, delta, and epsilon using the embryonic heart cell line, H9c2, and how this may be linked to changes in IGF1 signal transduction. Western blot analyses of H9c2 protein preparations demonstrate that there are significant increases in the total protein levels of PKC delta and epsilon after 4 days exposure to alcohol, and similar increases were found after 2 and 6 days exposure. In addition, there was a significant increase in PKC delta and epsilon in the membranal fractions and a decrease in the cytosolic fractions. No change was found in the expression or activity levels for PKC alpha. Chronic-alcohol exposure (100 mM, 4 days) increased the basal tyrosine kinase activity of the IGF1 receptor (IGF1R), and altered its rate of activation. Chronic-alcohol exposure also reduced the rate of Erk1/Erk2 activation by IGF1. Chronic alcohol blocked the proliferative effects of IGF1 on cell growth and reduced cell viability both in the presence and absence of IGF1, and this alcohol-induced reduction in cell viability was blocked using siRNA to inhibit PKC delta. In addition, a reduction in the amount of myosin light chain 2 was found in the alcohol-exposed cells. In conclusion, chronic alcohol alters PKC delta and epsilon expression and activity, and suppresses the IGF1 signaling pathway in embryonic heart cell culture. Blockage of PKC delta expression using siRNA inhibits the suppressive effects of alcohol on cell viability. PMID- 17127137 TI - Metabolic basis of ethanol-induced hepatic and pancreatic injury in hepatic alcohol dehydrogenase deficient deer mice. AB - Alcoholic liver disease (ALD) and alcoholic pancreatitis (AP) are major diseases causing high mortality and morbidity among chronic alcohol abusers. Neutral lipid accumulation (steatosis) is an early stage of ALD or AP and progresses to inflammation and other advanced stages of diseases in a subset of chronic alcohol abusers. However, the mechanisms of alcoholic steatosis leading to ALD and AP are not well understood. Chronic alcohol abuse impairs hepatic alcohol dehydrogenase (ADH, a major enzyme involved in ethanol oxidative metabolism) and facilitates nonoxidative metabolism of ethanol to fatty acid ethyl esters (FAEEs, nonoxidative metabolites of ethanol). These esters are implicated in the pathogenesis of various alcoholic diseases and shown to cause hepatocellular and pancreatitis-like injury. Ethanol exposure is known to increase synthesis of FAEEs by several-fold in the livers and pancreata of rats pretreated with hepatic ADH inhibitor. Therefore, studies were undertaken to evaluate hepatocellular and pancreatic injury in hepatic ADH-deficient (ADH(-)) deer mice versus ADH-normal (ADH(+)) deer mice fed ethanol (4% wt/vol) via Lieber-DeCarli liquid diet for 60 days. A significant mortality was found in ethanol-fed ADH(-) deer mice (11 out of 18) versus ADH(+) deer mice (1 out of 16); most of the deaths occurred during the first 2 weeks of ethanol exposure. The surviving animals, sacrificed at the end of 60th day, showed distinct changes in hepatic and pancreatic histology and several-fold increases in nonoxidative metabolism of ethanol in ethanol-fed ADH( ) versus ADH(+) deer mice. Extensive vacuolization with displacement or absence of nucleus in some hepatocytes, and significant increase in hepatic neutral lipids were found in ethanol-fed ADH(-) versus ADH(+) deer mice. Ultrastructural changes showed perinuclear space, edema, presence of apoptotic bodies and disintegration, and/or dilatation of endoplasmic reticulum (ER) in the pancreata of ethanol-fed ADH(-) deer mice. FAEE levels were significantly higher in ADH(-) versus ADH(+) deer mice, approximately four-fold increases in the livers and seven-fold increases in the pancreata. Ethyl esters of oleic, linoleic, and arachidonic acids were the major FAEEs detected in ethanol-fed groups. The role of FAEEs in pancreatic lysosomal fragility is reflected by higher activity of cathepsin B (five-fold) in ethanol-fed ADH(-) versus ADH(+) deer mice. Although the present studies clearly indicate a metabolic basis of ethanol-induced hepatic and pancreatic injury, detailed dose- and time-dependent toxicity studies in this ADH(-) deer mouse model could reveal further a better understanding of mechanism(s) of ethanol-induced hepatic and pancreatic injuries. PMID- 17127140 TI - Thyrotoxicosis and thyroid storm. AB - Thyroid storm represents the extreme manifestation of thyrotoxicosis as a true endocrine emergency. Although Grave's disease is the most common underlying disorder in thyroid storm, there is usually a precipitating event or condition that transform the patient into life-threatening thyrotoxicosis. Treatment of thyroid storm involves decreasing new hormone synthesis, inhibiting the release of thyroid hormone, and blocking the peripheral effects of thyroid hormone. This multidrug, therapeutic approach uses thionamides, iodine, beta-adrenergic receptor antagonists, corticosteroids in certain circumstances, and supportive therapy. Certain conditions may warrant the use of alternative therapy with cholestyramine, lithium carbonate, or potassium perchlorate. After the critical illness of thyroid storm subsides, definitive treatment of the underlying thyrotoxicosis can be planned. PMID- 17127141 TI - Myxedema coma. AB - Myxedema coma is the term given to the most severe presentation of profound hypothyroidism and is often fatal in spite of therapy. Decompensation of the hypothyroid patient into a coma may be precipitated by a number of drugs, systemic illnesses (eg, pneumonia), and other causes. It typically presents in older women in the winter months and is associated with signs of hypothyroidism, hypothermia, hyponatremia, hypercarbia, and hypoxemia. Treatment must be initiated promptly in an intensive care unit setting. Although thyroid hormone therapy is critical to survival, it remains uncertain whether it should be administered as thyroxine, triiodothyronine, or both. Adjunctive measures, such as ventilation, warming, fluids, antibiotics, pressors, and corticosteroids, may be essential for survival. PMID- 17127142 TI - Emergencies caused by pheochromocytoma, neuroblastoma, or ganglioneuroma. AB - Pheochromocytoma may lead to important emergency situations, ranging from cardiovascular emergencies to acute abdomen and multiorgan failure. It is vital to think about this disease in any emergency situation when conventional therapy fails to achieve control or symptoms occur that do not fit the initial diagnosis. The importance of keeping this diagnosis in minds is underscored by the fact that, in 50% of pheochromocytoma patients, the diagnosis is initially overlooked. Two other tumors of the sympathetic nervous system, neuroblastoma and ganglioneuroma, are less commonly associated with emergency conditions. If they occur, they are often linked to catecholamine excess, paraneoplastic phenomena, or local tumor mass effect. PMID- 17127143 TI - Hyperglycemic crises in diabetes mellitus: diabetic ketoacidosis and hyperglycemic hyperosmolar state. AB - Diabetic ketoacidosis (DKA) and hyperglycemic hyperosmolar state (HHS) potentially are fatal but largely preventable acute metabolic conditions of uncontrolled diabetes, the incidence of which continues to increase. Mortality from DKA has declined remarkably over the years because of better understanding of its pathophysiology and treatment. The mortality rate of HHS remains alarmingly high, however, owing to older age and mode of presentation of patients and associated comorbid conditions. DKA and HHS also are economically burdensome; therefore, any resources invested in their prevention would be rewarding. PMID- 17127144 TI - Hypoglycemia. AB - Under physiologic conditions, glucose plays a critical role in providing energy to the central nervous system. A precipitous drop in the availability of this substrate results in dramatic symptoms that signal a medical emergency and warrant immediate therapy aimed at restoring plasma glucose to normal levels. A systemic approach to the differential diagnosis is useful in identifying the cause of hypoglycemia. Once established, a specific and/or definitive intervention that addresses that underlying problem can be implemented. In most cases, this systemic approach to diagnosis and therapy is rewarded with a good outcome for the patient. PMID- 17127145 TI - Acute adrenal insufficiency. AB - Adrenal insufficiency is a rare disorder, usually with gradually evolving clinical symptoms and signs. Occasionally, an acute adrenal insufficiency crisis can become a life-threatening condition because of acute interruption of a normal or hyperfunctioning adrenal or pituitary gland or sudden interruption of a adrenal replacement therapy. Acute stress situations can aggravate the symptomatology. A simple strategy or diagnostic screening and early intervention with sodium chloride-containing fluids and hydrocortisone should be widely implemented for cases with suspicion of an acute Addison disease crisis. In contrast, the chronic replacement dosage for patients with adrenal insufficiency should be as low as possible with clear instructions for dosage adjustments in case of stress or acute emergencies. PMID- 17127146 TI - The dynamic neuroendocrine response to critical illness. AB - The severity of striking alterations in the hypothalamic-anterior pituitary peripheral hormone axes, which are the hallmark of severity of critical illness, is associated with a high risk for morbidity and mortality. Most attempts to correct the hormone balance are ineffective or harmful because of lack of pathophysiologic understanding. Extensive research has provided more insight in the biphasic neuroendocrine response to critical illness: the acute phase is characterized by an actively secreting pituitary but low peripheral effector hormone levels. In contrast, in prolonged critical illness, uniform suppression of the neuroendocrine axes, predominantly of hypothalamic origin, contributes to low serum levels of the respective target-organ hormones. PMID- 17127147 TI - Changes within the growth hormone/insulin-like growth factor I/IGF binding protein axis during critical illness. AB - Interest in the somatotropic axis,with its complex network of interactions, during critical illness arose only a few decades ago. Te distinguishing neuroendocrine features of prolonged critical illness were not differentiated from those during the acute phase until the early 1990s. This incomplete understanding of the somatotropic axis contributed to some disastrous results, such as the multicenter growth hormone trial. The goal of stimulating the somatotropic axis without a proper preceding neuroendocrine diagnosis should be held obsolete. Moreover, the fascinating link between regulators of carbohydrate metabolism, such as insulin and insulin-like growth factor I, and the somatotropic axis may lead to future therapeutic possibilities. PMID- 17127148 TI - Changes within the thyroid axis during the course of critical illness. AB - This article reviews the mechanisms behind the observed changes in plasma thyroid hormone levels in the acute phase and the prolonged phase of critical illness. It focuses on the neuroendocrinology of the low triiodothyronine syndrome and on thyroid hormone metabolism by deiodination and transport. PMID- 17127149 TI - The hypothalamic-pituitary-adrenal axis in critical illness. AB - The hypothalamic-pituitary-adrenal response to stress is a dynamic process. The homeostatic corrections that have emerged in the course of human evolution to cope with the catastrophic events during critical illness involve a complex multisystem endeavor. Although the repertoire of endocrine changes has been probed in some detail, discerning the vulnerabilities and failures of this system is far more challenging. One of the most controversially debated topics in the current literature is the characterization and optimal treatment of allegedly inadequate adaptations of the hypothalamic-pituitary-adrenal axis during critical illness. This outline attempts to touch briefly some of the debated issues, stir the discussion, and thereby contribute to resolving the dispute. PMID- 17127150 TI - Catecholamines and vasopressin during critical illness. AB - This article summarizes the effects of catecholamines and vasopressin on the cardiovascular system, focusing on their metabolic and immunologic properties. Particular attention is dedicated to the septic shock condition. PMID- 17127151 TI - Diabetes of injury: novel insights. AB - Critically ill patients usually develop hyperglycemia, a condition referred to as "diabetes of injury." More and more evidence argues against the concept that this is an adaptive beneficial response. Indeed, the development of hyperglycemia seems to be detrimental for the outcome of critically ill patients, because maintenance of normoglycemia with intensive insulin therapy prevents morbidity and reduces mortality of critically ill patients to a large extent. The mechanisms underlying these clinical benefits are being studied further. PMID- 17127152 TI - Disorders of body water homeostasis in critical illness. AB - Disorders of sodium and water homeostasis are among the most commonly encountered disturbances in the critical care setting, because many disease states cause defects in the complex mechanisms that control the intake and output of water and solute. Because body water is the primary determinant of extracellular fluid osmolality, disorders of body water balance can be categorized into hypoosmolar and hyperosmolar disorders depending on the presence of an excess or a deficiency of body water relative to body solute. Because the main constituent of plasma osmolality is sodium, hypoosmolar and hyperosmolar disease states are generally characterized hy hyponatremia and hypernatremia, respectively. After a brief review of normal water metabolism, this article focuses on the diagnosis and treatment of hyponatremia and hypernatremia in the critical care setting. PMID- 17127155 TI - What is a residency? What is a resident? AB - Residency programs exist to train the next generation of podiatric physicians. A brief review of residency programs and what constitutes a "resident" is offered in a concise format. PMID- 17127156 TI - Evidence-based medicine in podiatric residency training. AB - Evidence-based medicine--the use of "current best evidence" in making medical decisions--is becoming the standard system for health care delivery. In applying evidence-based medicine, the podiatric profession lags behind in several key ways. Major problems include a scarcity of high-level evidence for most podiatric issues and a lack of database resources that summarize the evidence for easy access. This lack of resources complicates the use of evidenced-based medicine as a teaching tool in podiatric residencies. Other complicating factors include high patient volumes, the need for multiple training sites, and limited control over faculty who are mostly volunteers with little or no formal training in educational methods. PMID- 17127157 TI - Conducting a meaningful interview: making the most of a golden opportunity. AB - The interview is an integral part of the residency selection process, and is a golden opportunity for you and the student. If you are willing to consider certain pre-interview decisions and structure your interview in a way that allows you to learn more about a candidate than just their academic accomplishments, there is an excellent chance that you will select and match with candidates who best complement your learning environment. PMID- 17127158 TI - Teaching during rounds. AB - Teaching rounds are a vital part of every residency program. Patient rounds provide an avenue to incorporate evidence-based medicine, current literature, and personal experiences into daily teaching while allowing residents to experience the doctor-patient relationship. Although time consuming, daily in-patient rounds are invaluable to any physician's training. This article discusses how to conduct well-organised, efficient patient rounds with maximum learning benefits for the residents. The daily protocol followed by the Podiatry Service at University Hospital, University of Medicine and Dentistry of New Jersey is also described. Conducting beneficial teaching rounds is an important aspect of a residency program when developing highly competent physicians. PMID- 17127159 TI - Training a competent surgeon. AB - Residency education is evolving into a competency model. Defining and evaluating competence are new paradigms that must be rapidly inserted into residency programs to satisfy residency requirements. PMID- 17127160 TI - The successful journal club. AB - Journal club is a structured meeting that is required at a residency program, which is designated to train residents in the necessary skills to evaluate and apply scientific literature critically to clinical decision making. A successful journal club is one in which residents develop competency in evaluating the scientific literature for evidence-based answers that can be applied to clinical questions. The objective in establishing a successful journal club is to build a forum for residents to formulate answers to their clinical questions through the development of essential critical appraisal skills. This article discusses the setting, format, content, and purpose of a successful journal club. PMID- 17127161 TI - The use of patient simulators in podiatric medical/surgical education. AB - Patient simulators have made their mark in medical education, and will be a permanent, integral part in the training of medical specialties. Their scope will always be increasing and changing as technology keeps improving the quality and the sophistication of the simulators. Studies have shown that patient care and treatment has improved with the use of simulators, allowing true mental and physical confidence of the practioners before and during patient care. The role of patient simulators has increased in podiatric education, and will now become an integral part during podiatric college, podiatric residency, and most likely as postgraduate continuing medical education credits. PMID- 17127162 TI - The 360 degrees evaluation. AB - The 360 degrees evaluation refers to resident assessment by all persons in the resident's sphere of influence. Although most think of the 360 degrees evaluation as a single tool used by all nonfaculty personnel, it is actually an assessment system. Therefore, all evaluators, including faculty, and all tools used in the assessment of residents comprise the 360 degrees evaluation. The goal of the 360 degrees evaluation is to accurately assess resident performance using tools that are reliable and valid so that competence can be demonstrated and programmatic needs identified. This article provides insights into the evaluation process through guidelines, practical advice, and examples from the field. PMID- 17127163 TI - A practical approach to research during residency training. AB - This article describes a basic model for clinical research in a 3-year residency training program. Obstacles to research along with practical solutions are presented. Performing clinical research should be an integral part of residency training with publication being the ultimate goal for every resident research project. PMID- 17127164 TI - Competencies as an evaluation tool. AB - Competency-based resident education focuses on resident performance (learning outcomes) in reaching specific competencies (goals and objectives of the curriculum). The seven competencies that are outlined in the Council on Podiatric Medical Education 320 document are organizing principles for a podiatric residency curriculum. Through the use of the competencies, the resident evaluation can be constructed to best determine the need for resident intervention as well as program improvements. PMID- 17127165 TI - In-training examinations. AB - In-training are one way to evaluate the residents during their training. Currently, the American Board of Podiatric Surgery supplies the examination. Various residency directors were questioned regarding its use and importance. PMID- 17127166 TI - [99mTc]HYNIC-RGD for imaging integrin alphavbeta3 expression. AB - There has been increasing interest in peptides containing the Arg-Gly-Asp (RGD) sequence for targeting of alpha(v)beta(3) integrins to image angiogenesis. [(18)F]Galacto-RGD has been successfully used for positron emission tomography applications in patients. Here we report on the preclinical characterization of a (99m)Tc-labeled derivative for single-photon emission computed tomography. c(RGDyK) was derivatized with HYNIC at the amino group of the lysine [c(RGDyK(HYNIC)) or HYNIC-RGD]. (99m)Tc labeling was performed using coligands (tricine and EDDA), as well as (99m)Tc(CO)(3)(H(2)O)(3). Radiolabeled peptides were characterized with regard to lipophilicity, protein binding and stability in buffer, serum and tissue homogenates. Integrin receptor activity was determined in internalization assays using alpha(v)beta(3)-receptor-positive M21 and alpha(v)beta(3)-receptor-negative M21L melanoma cells. Biodistribution was evaluated in normal and nude mice bearing M21, M21L and small cell lung tumors. HYNIC-RGD could be labeled at high specific activities using tricine, tricine trisodium triphenylphosphine 3,3',3''-trisulfonate (TPPTS), tricine-nicotinic acid (NA) or EDDA as coligands. [(99m)Tc]EDDA/HYNIC-RGD, [(99m)Tc]tricine TPPTS/HYNIC-RGD and [(99m)Tc]tricine-NA/HYNIC-RGD showed protein binding (<5%) considerably lower than [(99m)Tc](CO)(3)/HYNIC-RGD and [(99m)Tc]tricine/HYNIC RGD. [(99m)Tc]EDDA/HYNIC-RGD revealed high in vitro stability accompanied by low lipophilicity with a log P value of -3.56, comparable to that of [(18)F]Galacto RGD. In M21 cells for this compound, the highest level of specific and rapid cell uptake (1.25% mg protein(-1)) was determined. In vivo, rapid renal excretion, low blood retention, low liver and muscle uptakes and low intestinal excretion 4 h postinjection were observed. Tumor uptake values were 2.73% ID/g in M21 alpha(v)beta(3)-receptor-positive tumors versus 0.85% ID/g in receptor-negative tumors 1 h postinjection. Small cell lung tumors could be visualized using gamma camera imaging. [(99m)Tc]EDDA/HYNIC-RGD shows encouraging properties to target alpha(v)beta(3) receptors in vivo with high stability and favorable pharmacokinetics. Tumor uptake studies showed specific targeting of alpha(v)beta(3)-receptor-positive tumors with tumor-to-organ ratios comparable to those of [(18)F]Galacto-RGD. PMID- 17127167 TI - Synthesis and biological evaluation of potent alphavbeta3-integrin receptor antagonists. AB - INTRODUCTION: alpha(v)beta(3) Integrin is expressed in sprouting endothelial cells in growing tumors, whereas it is absent in quiescent blood vessels. In addition, various tumor cell types express alpha(v)beta(3) integrin. alpha(v)beta(3) Integrin, a transmembrane heterodimeric protein, binds to the arginine-glycine-aspartic acid (RGD) amino acid sequence of extracellular matrix proteins such as vitronectin and plays a pivotal role in invasion, proliferation and metastasis. Due to the selective expression of alpha(v)beta(3) integrin in tumors, radiolabeled RGD peptides and peptidomimetics are attractive candidates for tumor targeting. METHODS: A cyclic RGD peptide, a peptoid-peptide hybrid, an all-peptoid and a peptidomimetic compound were synthesized, conjugated with 1,4,7,10-tetraazadodecane-N,N',N'',N'''-tetraacetic acid (DOTA) and radiolabeled with (111)In. Their in vitro and in vivo alpha(v)beta(3)-binding characteristics were determined. RESULTS: IC(50) values were 236 nM for DOTA-E-c(RGDfK), 219 nM for DOTA-peptidomimetic, >10 mM for DOTA-all-peptoid and 9.25 mM for the peptoid peptide hybrid DOTA-E-c(nRGDfK). (111)In-labeled compounds, except for [(111)In]DOTA-all-peptoid, showed specific uptake in human alpha(v)beta(3) expressing tumors xenografted in athymic mice. Tumor uptake for [(111)In]DOTA-E c(RGDfK) was 1.73+/-0.4% ID/g (2 h postinjection) and that of [(111)In]DOTA peptidomimetic was 2.04+/-0.3% ID/g. Tumor uptake for the peptoid-peptide hybrid [(111)In]DOTA-E-c(nRGDfK) was markedly lower (0.45+/-0.07% ID/g). The all-peptoid [(111)In]DOTA-E-c(nRGnDnFnK) did not show specific uptake in tumors (0.11+/-0.04% ID/g). CONCLUSIONS: The peptidomimetic compound and the cyclic RGD peptide have a high affinity for alpha(v)beta(3) integrin, and these compounds have better tumor targeting characteristics than the peptoid-peptide hybrid and the all-peptoid. PMID- 17127168 TI - Radioiodinated VEGF to image tumor angiogenesis in a LS180 tumor xenograft model. AB - INTRODUCTION: Angiogenesis is essential for tumor growth or metastasis. A method involving noninvasive detection of angiogenic activity in vivo would provide diagnostic information regarding antiangiogenic therapy targeting vascular endothelial cells as well as important insight into the role of vascular endothelial growth factor (VEGF) and its receptor (flt-1 and KDR) system in tumor biology. We evaluated radioiodinated VEGF(121), which displays high binding affinity for KDR, and VEGF(165), which possesses high binding affinity for flt-1 and low affinity for KDR, as angiogenesis imaging agents using the LS180 tumor xenograft model. METHODS: VEGF(121) and VEGF(165) were labeled with (125)I by the chloramine-T method. Biodistribution was observed in an LS180 human colon cancer xenograft model. Additionally, autoradiographic imaging and immunohistochemical staining of tumors were performed with (125)I-VEGF(121). RESULTS: (125)I VEGF(121) and (125)I-VEGF(165) exhibited strong, continuous uptake by tumors and the uterus, an organ characterized by angiogenesis. (125)I-VEGF(121) uptake in tumors was twofold higher than that of (125)I-VEGF(165) (9.12+/-98 and 4.79+/ 1.08 %ID/g at 2 h, respectively). (125)I-VEGF(121) displayed higher tumor to nontumor (T/N) ratios in most normal organs in comparison with (125)I-VEGF(165). (125)I-VEGF(121) accumulation in tumors decreased with increasing tumor volume. Autoradiographic and immunohistochemical analyses confirmed that the difference in (125)I-VEGF(121) tumor accumulation correlated with degree of tumor vascularity. CONCLUSION: Radioiodinated VEGF(121) is a promising tracer for noninvasive delineation of angiogenesis in vivo. PMID- 17127169 TI - Blood flow dependence of the intratumoral distribution of peripheral benzodiazepine receptor binding in intact mouse fibrosarcoma. AB - The intratumoral distribution of [(11)C]AC-5216 binding, a novel peripheral benzodiazepine receptor (PBR) ligand, was examined by autoradiography both in vitro and in vivo using a murine fibrosarcoma model. The regional distribution of [(11)C]AC-5216 in a tumor in vivo was significantly heterogeneous; the uptake of [(11)C]AC-5216 was comparatively higher in the outer rim of the tumor and was lower in the central area. In contrast, the images obtained following the injection of [(11)C]AC-5216 with a large amount of nonlabeled PK11195 showed a relatively homogeneous distribution, suggesting that [(11)C]AC-5216 uptake represented specific binding to PBRs. In vitro autoradiograms of [(11)C]AC-5216 binding were also obtained using the section of the fibrosarcoma that was the same as that used to examine in vivo binding. In vitro autoradiographic binding images showed homogeneous distribution, and significant discrepancies of the intratumoral distribution of [(11)C]AC-5216 were observed between in vivo and in vitro images. The in vivo images of [(11)C]AC-5216 uptake, compared with those of [(14)C]iodoantipyrine uptake, obtained by dual autoradiography to evaluate the influence of blood flow revealed the similar intratumoral distributions of both tracers. These results indicate that the delivery process from the plasma to the tumor might be the rate-limiting step for the intratumoral distribution of PBR binding in vivo in a fibrosarcoma model. PMID- 17127170 TI - Effect of hypoxia on the uptake of [methyl-3H]choline, [1-14C] acetate and [18F]FDG in cultured prostate cancer cells. AB - INTRODUCTION: Choline, acetate and glucose ([2-(18)F]fluoro-2-deoxyglucose, [(18)F]FDG) analogs are under investigation as positron emission tomography (PET) tracers for the imaging of prostate cancer; however, their response to tumor hypoxia has not been clarified. METHODS: The uptake of [methyl-(3)H]choline, [1 (14)C]acetate and [(18)F]FDG was monitored in androgen-independent PC-3 cells and androgen-sensitive LNCaP cells under aerobic or anoxic conditions. The effect of androgen depletion was also examined. RESULTS: PC-3 cells exhibited aerobic choline and acetate uptake five to six times higher than FDG uptake, whereas LNCaP cells showed choline uptake 2.2-fold higher than acetate uptake and 10-fold higher than FDG uptake. After 4 h of anoxia, PC-3 cells showed an 85% increase in FDG uptake, a 15% decrease in choline uptake and a 36% increase in acetate uptake, whereas LNCaP cells showed a 212% increase in FDG uptake, a 28% decrease in choline uptake and no change in acetate uptake. Androgen depletion resulted in a marked decrease in the uptake of all tracers in LNCaP cells but no changes in PC-3 cells. CONCLUSION: In aerobic conditions, both PC-3 and LNCaP cells exhibited an order of uptake preference as follows: choline>acetate>FDG. In hypoxic cells, the order is reversed, reflecting diverse biochemical responses to hypoxia. These findings may help to explain PET imaging findings of the diverse responses of these tracers in different stages and locations of prostate cancer. Androgen depletion markedly suppressed the uptake of all three tracers in LNCaP cells, which suggests the potential for underestimation of the disease state when PET imaging is performed subsequent to antiandrogen therapy. PMID- 17127171 TI - Evaluation of [14C]phenylacetate as a prototype tracer for the measurement of glial metabolism in the rat brain. AB - [(14)C]Phenylacetate was designed as a prototype tracer for the measurement of glial metabolism, and its potency in comparison with that of [(14)C]acetate was evaluated in this study. Normal rats were intravenously injected with [(14)C]phenylacetate or [(14)C]acetate, and radioactivity concentrations were measured in the plasma, cerebral cortex, cerebellum and peripheral tissues by dissection method. In addition, [(14)C]phenylacetate uptake in the rat brain was compared by autoradiography with that of [(14)C]acetate following the injection of fluorocitrate, a selective glial toxin, into the brain. [(14)C]Phenylacetate was rapidly taken up into the brain and was retained at high levels up to 20 min postadministration. The levels of [(14)C]phenylacetate in the cerebral cortex were about threefold higher than those of [(14)C]acetate at 1 min postinjection. Microinjection of fluorocitrate into the right striatum resulted in a significant decrease of the uptake of both [(14)C]phenylacetate and [(14)C]acetate into the right striatum. Radiochemical analysis confirmed the rapid hydrolysis of [(14)C]phenylacetate in the rat brain, with less than 20% of radioactivity representing unmetabolized [(14)C]phenylacetate at 1 min postinjection. These results suggest that [(14)C]phenylacetate is rapidly taken up into the brain and is hydrolyzed and converted to [(14)C]acetate. [(14)C]Phenylacetate may have the potential to serve as a tracer for the measurement of glial metabolism in an intact brain. PMID- 17127172 TI - In vitro and in vivo characterization of 177Lu-huA33: a radioimmunoconjugate against colorectal cancer. AB - INTRODUCTION: The humanized monoclonal antibody A33 (huA33) is a potential targeting agent against colorectal carcinoma since the A33 antigen is highly and homogenously expressed in >95% of all colorectal cancers, both primary tumors and metastases. The aim of this study was to determine the biodistribution and tumor targeting ability of (177)Lu-labeled huA33. METHODS: huA33 was labeled with the beta-emitting therapeutic nuclide (177)Lu using the chelator CHX-A"-DTPA, and the properties of the (177)Lu-CHX-A"-huA33 ((177)Lu-huA33) conjugate was determined both in vitro and in vivo in a biodistribution study in nude mice xenografted with colorectal SW1222 tumor cells. RESULTS: The (177)Lu-huA33 conjugate bound specifically to colorectal cancer cells in vitro (with a K(D) value of 2.3+/-0.3 nM, determined by a saturation assay) and in vivo. The tumor uptake of (177)Lu huA33 was very high, peaking at 134+/-21%ID/g 72 h postinjection (pi). Normal tissue uptake was low; radioactivity concentration in blood (which had the second highest radioactivity concentration) was lower than in tumor at all time points studied (8 h to 10 days). The tumor-to-blood ratio increased with time, reaching 70+/-30, 10 days pi. Throughout the study, the uptake of (177)Lu in bone (known to accumulate free (177)Lu) was low, and the fraction of protein-bound (177)Lu in plasma samples was high (95% to 99%). This indicates high stability of the (177)Lu-huA33 conjugate in vivo. CONCLUSION: The (177)Lu-huA33 conjugate shows a very favorable biodistribution, with an impressively high tumor uptake and high tumor-to-organ ratios, indicating that the conjugate may be suitable for radioimmunotherapy of colorectal cancer. PMID- 17127173 TI - Correlations between 18F-FDG uptake by bone marrow and hematological parameters: measurements by PET/CT. AB - The aim of this study was to investigate the correlations between (18)F-FDG uptake by bone marrow and various hematological parameters. Forty-eight patients who underwent FDG-PET/CT studies and also received hematological examinations within 5 days before or after the PET study were included in this study. All patients had not received chemotherapy. FDG uptake by bone marrow was measured as a standardized uptake value (SUV) on three-dimensional PET/CT fusion images, and the uptake ratio (UR) of the SUV of bone marrow to the SUV of longitudinal dorsal muscle was calculated. The correlations between the SUV and the UR of bone marrow and various hematological parameters were evaluated. Bone marrow FDG uptake was strongly correlated with the white blood cell counts but was not significantly correlated with the red blood cell and platelet counts. The neutrophil count was significantly correlated with bone marrow FDG uptake but the lymphocyte count was not. FDG uptake by bone marrow was specifically correlated with the neutrophil count, suggesting that the FDG uptake by bone marrow reflects marrow metabolism that is mainly regulated by granulocyte progenitors and stimulated by endogenous hematopoietic growth factors. They may also be helpful in interpreting PET images, especially for diagnosing bone marrow involvement by malignancy. PMID- 17127174 TI - The tyrosine kinase inhibitor PD153035: implication of labeling position on radiometabolites formed in vitro. AB - INTRODUCTION: The epidermal growth factor receptor is highly expressed in several types of cancers. Molecules with high affinity to its intracellular tyrosine kinase domain are being developed as in vivo imaging probes. The 4 anilinoquinazoline PD153035 has promising in vitro and in vivo properties for development as a reversible radioligand. Labeling it with carbon-11 in either of its two methoxy positions can potentially give rise to different radiometabolites and, consequently, different imaging capabilities. An evaluation of the radiotracers' metabolism was needed to determine the potential significance of the labeling position. METHODS: PD153035 was labeled in the 6- and 7-O-methoxy positions by reacting the corresponding O-desmethyl precursors with [(11)C]methyl iodide. The two radiolabeled compounds were each incubated for 1 h with human and rat liver microsomes. At five time points, the radiolabeled metabolites were examined using radio-liquid chromatography. One metabolite was isolated and subjected to mass spectroscopic analysis. RESULTS: A major polar metabolite was obtained in all incubations. Its molecular weight was consistent with an addition of oxygen, and its fragmentation was consistent with an N-oxidation rather than an aromatic hydroxylation. Regioselective 7-O-dealkylation was also observed, albeit in substantial amounts only in the assay using human microsomes. CONCLUSIONS: Radiolabeling in the 7-O-methoxy position is advocated, since the labeled metabolites produced in the 7-O-demethylation are polar and probably rapidly cleared. The differences observed in the incubations with rat and human microsomes suggest that in vivo positron emission tomography studies with (11)C labeled PD153035 in rodents may not be directly predictive for studies in humans. PMID- 17127175 TI - Species differences in metabolites of PET ligands: serotonergic 5-HT1A receptor antagonists 3-trans-FCWAY and 3-cis-FCWAY. AB - Liquid chromatography/mass spectrometry (LC/MS), combined with commercially available hepatocytes, has become an indispensable tool in evaluating the presence of these metabolites in target tissues, especially in the brain. Results from in vitro metabolism studies using hepatocytes from different species can demonstrate species differences. Using these techniques, we evaluated the metabolic profile of 3-cis-FCWAY and 3-trans-FCWAY in rat, monkey and human hepatocytes. Hepatocytes were used to produce metabolites in vitro, and liquid chromatography/tandem mass spectrometry was used to identify these metabolites. We found that the metabolic profiles of rat, monkey and human hepatocytes differ dramatically. In rats, aromatic ring oxidation was the major metabolic pathway for both 3-cis-FCWAY and 3-trans-FCWAY; 3-trans-FCWAY had more metabolites (cyclohexane ring oxidation) than 3-cis-FCWAY. In humans, hydrolysis of amide linkage was the major metabolic pathway. In monkeys, both pathways (oxidation and amide hydrolysis) were found in the metabolites. We also found that 3-cis-FCWAY had the slowest defluorination of FCWAY analogues in all species. PMID- 17127176 TI - In vitro and in vivo evaluation of [11C]MPEPy as a potential PET ligand for mGlu5 receptors. AB - Excessive activation via the metabotropic glutamate receptor subtype 5 (mGluR(5)) has been implicated in depression, neuropathic pain and other psychiatric, neurological and neurodegenerative diseases. A mGluR(5) radioligand for in vivo quantification by positron emission tomography (PET) would facilitate studies of the role of this receptor in disease and treatment. 3-Methoxy-5-pyridin-2 ylethynylpyridine (MPEPy), a selective and high-affinity antagonist at the mGluR(5) receptor was selected as a candidate ligand; a recent publication by Yu et al. [Nucl Med Biol 32 (2005) 631-640] presented initial micro-PET results for [(11)C]MPEPy with enthusiasm. Building on their efforts, we report as unique contributions (1) an improved chemical synthesis method, (2) the first data using human tissue, (3) phosphor images for rat brain preparations, (4) a novel comparison of anesthetic agents and (5) in vivo data in baboon. In vitro phosphor imaging studies of this ligand using human and rat brain tissue demonstrated high specific binding in the hippocampus, striatum and cortex with minimal specific binding in the cerebellum. In contrast, in vivo micro-PET studies in rats using urethane anesthesia, PET studies in baboons using isoflurane anesthesia and ex vivo micro-PET studies in unanesthetized rats each showed little specific binding in the brain. Despite the promising in vitro results, the low signal-to-noise ratio found in vivo does not justify the use of [(11)C]MPEPy as a PET radiotracer in humans. PMID- 17127177 TI - Differential uptake of [18F]FET and [3H]l-methionine in focal cortical ischemia. AB - Amino acids such as [(11)C-methyl]l-methionine are particularly useful in brain tumor diagnosis, but unspecific uptake (e.g., in cerebral ischemia) has been reported. O-(2-[(18)F]fluoroethyl)-l-tyrosine ([(18)F]FET) shows a clinical potential similar to that of l-methionine (MET) in brain tumor diagnosis but is applicable on a wider clinical scale. The aim of this study was to evaluate the uptake of [(18)F]FET and [(3)H]MET in focal cortical ischemia in rats by dual tracer autoradiography. METHODS: Focal cortical ischemia was induced in 25 CDF rats using the photothrombosis (PT) model. At different time points up to 6 weeks after the induction of PT, [(18)F]FET and [(3)H]MET were injected intravenously. Additionally, contrast-enhanced magnetic resonance imaging (MRI) was performed in 10 animals. One hour after tracer injection, brains were cut in coronal sections and evaluated by dual-tracer autoradiography. Lesion-to-brain (L/B) ratios were calculated by dividing the maximal uptake in the lesion by the mean uptake in the brain. An L/B ratio of >2.0 was considered indicative of pathological uptake. Histological slices were stained by cresyl violet and supplemented by immunostainings for glial fibrillary acidic protein (GFAP) and CD68 in selected cases. RESULTS: A variably increased uptake of both tracers was observed in the PT lesion and its demarcation zone up to 7 days after PT for [(18)F]FET and up to 6 weeks for [(3)H]MET. The cutoff level of 2.0 was exceeded in 12/25 animals for [(18)F]FET and in 18/25 animals for [(3)H]MET. Focally increased tracer uptake matched contrast enhancement in MRI in 3/10 cases for [(18)F]FET and in 5/10 cases for [(3)H]MET. Immunohistochemical staining in lesions with differential uptake of [(18)F]FET and [(3)H]MET revealed that selective uptake of [(18)F]FET was associated with GFAP-positive astrogliosis while selective [(3)H]MET uptake correlated with CD68-positive macrophage infiltration. CONCLUSIONS: [(18)F]FET, like [(3)H]MET, may exhibit significant uptake in the periphery of cortical infarctions, which has to be considered in the differential diagnosis of unknown brain lesions. There are discrepancies between [(18)F]FET and [(3)H]MET uptake in the area of infarctions that appear to be caused by the preferential uptake of [(18)F]FET in reactive astrocytes versus the preferential uptake of [(3)H]MET in macrophages. PMID- 17127178 TI - Quantitative assessment of the atherosclerotic burden of the aorta by combined FDG-PET and CT image analysis: a new concept. AB - INTRODUCTION: Our objective was to develop a technique for quantifying the extent of atherosclerosis in the aorta by combining standard uptake values (SUVs) in the aortic wall with volumetric data provided by computed tomography (CT). METHODS: Eighteen patients who had both 18-fluorodeoxyglucose (FDG)-positron emission tomography (PET) and contrast-enhanced CT of the chest and abdomen were selected. All had homogeneous diffuse FDG wall uptake in four segments of the aorta. We divided the patients into three groups according to their age and measured FDG uptake in all four segments of the aorta by calculating the mean SUV for each segment. On each axial CT image, region-of-interest tracings along the inner and outer wall contours of the aorta were generated. The inner surface area was subtracted from the outer surface area. The net area values for each segment were subsequently multiplied by slice thickness to calculate arterial wall volume. By multiplying SUV with wall volume, we were able to calculate the atherosclerotic burden (AB) for each segment of the aorta. We compared the aortic wall volumes, SUVs and AB values in each arterial segment for each age group. RESULTS: In each aortic wall segment, AB values, SUVs and wall volumes increased with age (P<.05). CONCLUSION: AB can be used as an indicator of the extent of the atherosclerotic process in the aorta through the use of both metabolic and morphologic data provided by FDG-PET and CT, respectively. This may allow for the optimal screening, diagnosis and management of patients with this common and potentially lethal disorder. PMID- 17127179 TI - Evaluation of novel cationic 99mTc(I)-tricarbonyl complexes as potential radiotracers for myocardial perfusion imaging. AB - This report describes the evaluation of three cationic (99m)Tc(I)-tricarbonyl complexes--[(99m)Tc(CO)(3)(L)](+) (L=N-methoxyethyl-N,N-bis[2-(bis(3 ethoxypropyl)phosphino)ethyl]amine (ME-PNP), N-[15-crown-5)-2-yl]-N,N-bis[2 (bis(3-ethoxypropyl)phosphino)ethyl]amine (15C5-PNP) and N-[18-crown-6)-2-yl]-N,N bis[2-(bis(3-ethoxypropyl)phosphino)ethyl]amine (18C6-PNP))--as potential radiotracers for myocardial perfusion imaging. Biodistribution, imaging and metabolism studies were performed using Sprague-Dawley rats. It was found that bisphosphine ligands have a significant impact on the biodistribution characteristics and clearance kinetics of their cationic (99m)Tc(I)-tricarbonyl complexes. Among the three radiotracers evaluated in this study, [(99m)Tc(CO)(3)(15C5-PNP)](+) has a very high initial heart uptake and is retained in the rat myocardium for >2 h. It also shows rapid clearance from the liver and lungs. The heart/liver ratio of [(99m)Tc(CO)(3)(15C5-PNP)](+) is approximately 2.5 times better than that of (99m)Tc-sestamibi at 30 min postinjection. [(99m)Tc(CO)(3)(15C5-PNP)](+) is almost identical to (99m)TcN DBODC5 with respect to heart uptake, heart/lung ratio and heart/liver ratio. Results from metabolism studies show that there is no significant metabolism for [(99m)Tc(CO)(3)(15C5-PNP)](+) in the urine, but it does show a small metabolite peak (<10%) in the radio high-performance liquid chromatography chromatogram of the feces sample at 120 min postinjection. Results planar imaging studies demonstrate that [(99m)Tc(CO)(3)(15C5-PNP)](+) has a much better liver clearance profile than (99m)Tc-sestamibi and might give clinically useful images of the heart as early as 30 min postinjection. [(99m)Tc(CO)(3)(15C5-PNP)](+) is a very promising candidate for more preclinical evaluations in various animal models. PMID- 17127180 TI - Induction of apoptosis in human tumor cells after exposure to Auger electrons: comparison with gamma-ray exposure. AB - To clarify the contribution of apoptosis to cell death in four human solid tumor cell lines, clonogenic cell survival (indicator of radiosensitivity) and induction of caspase-3 (CASP-3)/caspase-3-like proteases (CASP-3LP) and the production of DNA fragmentation (markers for apoptosis) were studied in RKO, LS174T, MCF7 and TE671 cells exposed to DNA-incorporated Auger-electron-emitting (125)I (5-[(125)I]iodo-2'-deoxyuridine) or gamma-radiation. Clonogenic survival was assessed by colony-forming assay, CASP-3/CASP-3LP induction with a fluorogenic substrate and DNA fragmentation by ligation-mediated polymerase chain reaction. For (125)I, log dose-survival curves had no shoulder [high-linear energy-transfer (LET)-like] and decreased exponentially at different rates in various cell lines. Induction of CASP-3/CASP-3LP in radiosensitive RKO and LS174T cells was threefold greater than that in radioresistant TE671 and MCF7 cells. Nucleosomal laddering in (125)I-radiosensitive cell lines was dose-dependent, and no laddering was detected in radioresistant lines. For gamma-radiation, the survival curve for LS174T cells was monoexponential and that for the other lines exhibited a distinct shoulder (low-LET-like). The most radiosensitive cell line, LS174T, showed the highest induction of CASP-3/CASP-3LP, and the most radioresistant line, TE671, showed the lowest induction. Although DNA laddering was not detectable in TE671 cells, it was observed in other lines, being most prominent in LS174T cells. We conclude that apoptosis initiated by DNA incorporated (125)I is dose-dependent, correlates with cell radiosensitivity and takes place through a CASP-3-mediated pathway, whereas that after gamma irradiation probably occurs via a CASP-3-independent pathway and/or a CASP-3 mediated pathway and does not correlate with cell radiosensitivity. PMID- 17127181 TI - Fractionated radioimmunotherapy of intraperitoneally growing ovarian cancer in nude mice with 211At-MX35 F(ab')2: therapeutic efficacy and myelotoxicity. AB - OBJECTIVE: The aim of this study was to investigate the therapeutic efficacy and myelotoxicity during fractionated radioimmunotherapy of ovarian cancer in mice. The study was performed using the monoclonal antibody MX35 F(ab')(2) labeled with the alpha-particle emitter (211)At. METHODS: Animals were intraperitoneally inoculated with approximately 1x10(7) cells of the cell line NIH:OVCAR-3. Four weeks later, the mice were given the first treatment. Six groups of animals were intraperitoneally injected with approximately 800, 3x approximately 267, approximately 400, 3x approximately 133, approximately 50 or 3x approximately 17 kBq (211)At-MX35 F(ab')(2) (n=18 in each group). The second and third injections for Groups 2, 4 and 6 were given 4 and 8 days after the first injection, respectively. As controls, animals were treated with unlabeled MX35 F(ab')(2) (n=12). Eight weeks after the last injection, the animals were sacrificed and the presence of macro- and microscopic tumors and ascites was determined. Blood counts were determined for each mouse in Groups 1 and 2 before the first injection and 3, 7, 11, 15 and 23 days after the first injection. The calculation of the mean absorbed dose to the bone marrow was based on the ratio between the (211)At-activity concentration in bone and blood [i.e., the bone-to-blood ratio (BBLR)] as well as that between the (211)At-activity concentration in bone marrow and blood [i.e., the bone-marrow-to-blood ratio (BMBLR)] and the cumulated activity and absorbed fraction of the alpha-particles emitted by (211)At in the bone marrow. RESULTS: The tumor-free fractions of animals were 56% and 41% when treated with approximately 800 kBq and 3x approximately 267 kBq (211)At-MX35 F(ab')(2), respectively; 39% and 28% when treated with approximately 400 kBq and 3x approximately 133 kBq (211)At-MX35 F(ab')(2), respectively; and 17% and 22% when treated with approximately 50 kBq or 3x approximately 17 kBq (211)At-MX35 F(ab')(2), respectively. The nadir of the white blood cell (WBC) counts was decreased (from 46% to 19%, compared with the baseline WBC counts) and delayed (from Day 4 to Day 11 after the first injection) during the fractionated treatment compared with the single-dose treatment. The percentage of injected activity per gram (%IA/g) for blood, bone and bone marrow all peaked 6 h after injection at 13.80+/-1.34%IA/g, 4.00+/-0.69%IA/g and 8.28+/-1.38%IA/g, respectively. The BBLR and BMBLR were 0.20+/-0.04 and 0.58+/-0.01, respectively. The mean absorbed dose to bone marrow was approximately 0.4 Gy after intraperitoneally injecting approximately 800 kBq (211)At-MX35 F(ab')(2). CONCLUSION: No advantage was observed in the therapeutic efficacy of using a fractionated regimen compared with a single administration, with the same total amount of administered activity. Alleviation of the myelotoxicity was observed during the fractionated regimen in terms of decreased suppression and delayed nadir of the WBC counts. No thrombocytopenia was observed during either regimen. PMID- 17127183 TI - Gallstone disease: Epidemiology of gallbladder stone disease. AB - Gallstone disease is common: >700,000 cholecystectomies and costs of approximately 6.5 billion dollars annually in the U.S. The burden of disease is epidemic in American Indians (60-70%); a corresponding decrease occurs in Hispanics of mixed Indian origin. Ten to fifteen per cent of white adults in developed countries harbour gallstones. Frequency is further reduced in Black Americans, East Asia and sub-Saharan Africa. In developed countries, cholesterol gallstones predominate; 15% are black pigment. East Asians develop brown pigment stones in bile ducts, associated with biliary infection or parasites, or in intrahepatic ducts (hepatolithiasis). Certain risk factors for gallstones are immutable: female gender, increasing age and ethnicity/family (genetic traits). Others are modifiable: obesity, the metabolic syndrome, rapid weight loss, certain diseases (cirrhosis, Crohn's disease) and gallbladder stasis (from spinal cord injury or drugs like somatostatin). The only established dietary risk is a high caloric intake. Protective factors include diets containing fibre, vegetable protein, nuts, calcium, vitamin C, coffee and alcohol, plus physical activity. PMID- 17127184 TI - Gallstone disease. Pathogenesis of gallstones: A genetic perspective. AB - Cholelithiasis is one of the most prevalent gastroenterological diseases, imposing a huge economic burden on health-care systems. Gallbladder stones form when the concentration of cholesterol or bilirubin exceeds the solubility in the bile salt and phospholipid-rich bile. The physiology of biliary lipid secretion by a number of specialized transport proteins has recently been elucidated, and underlying genetic defects in these proteins have been identified as susceptibility factors for gallstone disease. Recent studies of identical twins and family strongly support the idea of a genetic component to gallstone disease. Epidemiological studies in high-risk populations indicate that gallstone formation is caused by multiple environmental influences and common genetic factors and their interactions. Monogenic subtypes of cholelithiasis, such as biliary lipid transporter deficiencies, appear to be rare. The characterization of lithogenic genes in knockout and transgenic mice, and the identification of many gallstone susceptibility loci in inbred mice, provide the basis for studies of the corresponding genes in patients with gallstones. The transfer of findings from mouse genetics to the bedside might lead to new strategies for individual risk assessment and reveal molecular targets for the development of new treatment strategies. PMID- 17127185 TI - Gallstone disease: Symptoms and diagnosis of gallbladder stones. AB - The clinical aspects and the diagnostic features of gallstone disease are described. The natural history of silent gallstones is overviewed, and the risk of developing symptoms and complications is also discussed. The importance of colicky pain as a specific gallstone symptom is highlighted, and the role of both laboratory tests and diagnostic investigations for differential diagnosis is discussed. Finally, we describe the diagnostic features of gallbladder stone disease, including indications, sensitivity, specificity, and limitations of different test investigations under special circumstances. PMID- 17127186 TI - Gallstone disease: Surgical aspects of symptomatic cholecystolithiasis and acute cholecystitis. AB - Indications for cholecystectomy are limited to clearly symptomatic gallstones. Relatively high failure rates of pain relief are probably caused by incorrect selection of patients for the operation. Contraindications for (laparoscopic) cholecystectomy are related to anaesthesiological considerations. Laparoscopic cholecystectomy was accepted by consensus as the gold standard within 5 years of its introduction. Nevertheless, both the classical open and small-incision cholecystectomies are safe alternatives, and superiority of any one of the three techniques over the others has not been proven. Primary outcome measures (mortality, complications, and symptom relief) seem to be equal for the three techniques. Acute cholecystitis is a complication of gallstones. Generally it is recommended that early cholecystectomy be performed, as delayed cholecystectomy is associated with longer total hospital stay and convalescence due to recurrent cholecystitis episodes. Cholecystostomy is an alternative technique for patients unfit for general anaesthesia. Reported complication and conversion rates in cholecystectomy for acute cholecystitis vary, but are higher than for symptomatic cholecystolithiasis. New developments--such as robot-assisted surgery--are expected to have a significant impact in the near future. PMID- 17127187 TI - Gallstone disease: Microlithiasis and sludge. AB - 'Sludge' is the solid material which results from the slow settling of particles dispersed in a liquid medium. Biliary sludge in the gallbladder can be detected by transabdominal ultrasonography, and the typical echoes derive mainly from pigment precipitates mixed with cholesterol crystals. A portion of biliary sludge contains comparatively large particles (1-3 mm) called microliths, the formation of which is an obligatory intermediate step in the development of all types of gallstone. Microlithiasis and sludge in bile may cause colicky pain, cholecystitis, cholangitis, and acute pancreatitis, and are thus of clinical relevance. In these patients treatment follows the guidelines of symptomatic gallstone disease, and strategies include long-term application of ursodeoxycholic acid, endoscopic papillotomy, or preferably laparoscopic cholecystectomy. PMID- 17127188 TI - Gallstone disease: Primary and secondary prevention. AB - Several risk factors for cholesterol gallstone formation in the general population have been identified. There is a strongly increased risk of gallstone disease during prolonged fasting, rapid weight loss, total parenteral nutrition, and somatostatin(-analogue) treatment. The annual risk of biliary colic and gallstone complications in asymptomatic gallstone carriers has been investigated sparsely. In asymptomatic and symptomatic gallstone carriers, treatment with the hydrophilic bile salt ursodeoxycholic acid (UDCA) has been claimed to reduce the risk of biliary colic and gallstone complications such as acute cholecystitis and acute pancreatitis. Also, prophylactic cholecystectomy could be beneficial in certain subgroups of asymptomatic gallstone carriers. However, randomized, double blind, placebo-controlled trials are lacking. In this review, strategies for the prevention of gallstone formation in the general population and in high-risk conditions are dealt with. Also, strategies for the prevention of biliary colic and gallstone complications in asymptomatic and symptomatic gallstone carriers are discussed. PMID- 17127189 TI - Gallstone disease: Epidemiology, pathogenesis, and classification of biliary stones (common bile duct and intrahepatic). AB - Gallstones are common in Western countries and Japan. Most gallstones are found in the gallbladder, but they sometimes pass through the cystic duct into extrahepatic and/or intrahepatic bile ducts to become bile-duct stones, causing conditions known as choledocholithiasis and hepatolithiasis. Some 10-15% of gallstone patients concomitantly suffer from bile-duct stones. Bile-duct stones can also be formed in the absence of gallbladder stones, and such primary bile duct stones are more common in East Asian countries than in the Western world. Thus pathogenesis of primary and secondary bile-duct stones is unlikely to be similar. Furthermore, the gallbladder stones are primarily cholesterol or black pigment stones, whereas most bile-duct stones are brown-pigment stones (calcium bilirubin stones). Thus, epidemiology, pathogenesis and classification of biliary stones are very likely to differ according to stone location (intrahepatic and/or extrahepatic bile duct). PMID- 17127190 TI - Gallstone disease: Symptoms, diagnosis and endoscopic management of common bile duct stones. AB - Bile duct stones (BDS) are often suspected on history and clinical examination alone but symptoms may be variable ranging from asymptomatic to complications such as biliary colic, pancreatitis, jaundice or cholangitis. The majority of BDS can be diagnosed by transabdominal ultrasound, computed tomography, endoscopic ultrasound or magnetic resonance cholangiography prior to endoscopic or laparoscopic removal. Approximately 90% of BDS can be removed following endoscopic retrograde cholangiography (ERC)+sphincterotomy. Most of the remaining stones can be removed using mechanical lithotripsy. Patients with uncorrected coagulopathies may be treated with ERC+pneumatic dilatation of the sphincter of Oddi. Shockwave lithotripsy (intraductal and extracorporeal) and laser lithotripsy have also been used to fragment large bile duct stones prior to endoscopic removal. The role of medical therapy in treatment of BDS is currently uncertain. This review focuses on the clinical presentation, investigation and current management of BDS. PMID- 17127191 TI - Gallstone disease. Management of common bile-duct stones and associated gallbladder stones: Surgical aspects. AB - For many years, open exploration of the common bile duct has been the treatment of choice for patients with common bile-duct stones. During recent decades endoscopic sphincterotomy has gained wide acceptance as an effective and less invasive alternative. After sphincterotomy, subsequent (laparoscopic) cholecystectomy is warranted in patients with gallbladder stones. This chapter will discuss whether sphincterotomy should be performed prior to, during or after cholecystectomy, and will also address the question of whether single-stage treatment by laparoscopic cholecystectomy and laparoscopic bile-duct exploration is in fact preferable. The rate of recurrent choledocholithiasis after endoscopic biliary sphincterotomy can reach more than 20%. This review focuses on the risk factors--delayed bile-duct clearance and bactobilia--that may lead to recurrent primary bile-duct stone formation. Underlying altered bile composition (relative phospholipid deficiency) should be recognised in a subgroup of patients. Identification of these risk factors may significantly affect treatment policy. PMID- 17127192 TI - Gallstone disease: Management of intrahepatic stones. AB - Hepatolithiasis (oriental cholangiohepatitis) has reportedly been endemic only in East Asia. The disease is now occasionally recognized in Western societies, especially in people who have lived in the Orient. Hepatolithiasis is characterized by its intractable nature and frequent recurrence, requiring multiple operative interventions, which is in distinct contrast to gallbladder stones. In addition to frequent cholangitis and chronic sepsis, it is widely known that longstanding intrahepatic stones lead to intrahepatic cholangiocarcinoma. Symptoms of hepatolithiasis include abdominal pain, jaundice and cholangitis. Pyogenic cholangitis due to strictures and hepatolithiasis tends to recur, and sometimes patients may present with liver abscesses. Radiological studies and percutaneous procedures are keys in the diagnosis and treatment of hepatolithiasis. Non-invasive imaging modalities such as ultrasonography (US), computed tomography (CT), and magnetic resonance imaging (MRI) accurately depict the normal anatomy and presence of intrahepatic stones. It should be stressed that each modality has its pros and cons, and imaging studies should be performed on the basis of understanding the pathophysiology. As the diagnostic role of magnetic resonance cholangiopancreatography (MRCP) evolves, the roles of both endoscopic retrograde cholangiopancreatography (ERCP) and percutaneous transhepatic cholangiography (PTC), and their most significant advantage, is primarily therapeutic with their ability to extract stones, biopsy intraductal lesions, and place stents easily. The primary goals of treatment are to eliminate attacks of cholangitis and to stop the progression of the disease (which leads to biliary cirrhosis). Surgery has a primary role in hepatolithiasis because hepatolithiasis tends to recur, so that multiple sessions of the endoscopic approach (i.e. two or three times a year) are often required. PTC is an alternative when surgical resection of the affected lobe is difficult. Techniques for lithotripsy, including shockwave and laser, can be applied in endoscopic sessions, offering a better chance of clearing the stones. PMID- 17127193 TI - Gallstone disease. Complications of bile-duct stones: Acute cholangitis and pancreatitis. AB - Gallstones are frequent in the Western world, with up to 10% of the general population affected. Gallstone prevalence is higher in the elderly and in women. Acute cholangitis and pancreatitis are the most serious complications of gallstones, with considerable morbidity and mortality. We discuss here clinical features, laboratory and radiological examinations, and treatment for gallstone cholangitis and pancreatitis. The diagnostic approach for acute 'idiopathic' pancreatitis is dealt with in some detail. Also, the role in pancreatitis of enteral nutrition, antibiotic prophylaxis, and the place of endoscopic retrograde cholangiography with papillotomy for biliary decompression is discussed in detail. PMID- 17127195 TI - Infection and musculoskeletal conditions: Infectious arthritis. AB - In contrast to the outstanding achievements made in therapy for autoimmune arthritides, not least rheumatoid arthritis, the pace of progress in therapy for infectious arthritis remains slow. This has primarily to do with the complex task of, on the one hand, killing the invading microorganisms and, on the other, to down-regulate the exaggerated immune response which participates in the microbial clearance but at the same time contributes to the tissue destruction. The use of experimental models of microbial arthritides has clarified several bacterial virulence factors as well as many haematopoietic cell types and their products that are involved in the pathogenesis of joint infection. Recent studies have documented that T-cell-mediated responses are not only prominent but also decisive with respect to disease sequelae. This chapter also reviews the primarily protective non-antigen-specific immune responsiveness to microbial agents, including the impact of neutrophils, complement system, and nitric oxide. The knowledge gained regarding microbial virulence factors and the host immune responses has prompted researchers to develop new strategies on how to interact in vivo with the infectious process. Some of these approaches are commented upon in this review: e.g. vaccination procedures to prevent septic arthritis and sepsis, as well as therapeutic procedures to minimize joint damage during an ongoing infection. PMID- 17127196 TI - Infection and musculoskeletal conditions: Prosthetic-joint-associated infections. AB - In patients with osteoarthritis or arthritis, prosthetic joint replacement is increasingly used to alleviate pain and to improve mobility. The most important risk factors are comorbidity and prior joint replacement (revision surgery). Diagnosis of prosthetic-joint-associated infection is difficult, because the infecting agent may be missed in synovial fluid due to its exclusive presence as a device-associated biofilm. Implant-associated infections are difficult to treat because of their resistance to natural host defence mechanisms and to most antibiotics. In staphylococcal implant-associated infections a rifampin combination should be used, because this drug has an excellent efficacy on surface-adhering microorganisms. Antimicrobial therapy must always be combined with the correct surgical treatment which is chosen according to an algorithm. The use of antibiotics during procedures with potential bacteraemia is controversial because evidence for its need is lacking. In contrast, during sepsis rapid antibiotic therapy is needed to prevent haematogenous seeding on the artificial joint. PMID- 17127197 TI - Infection and musculoskeletal conditions: Osteomyelitis. AB - Osteomyelitis can result from hematogenous or contiguous microbial seeding of the bone. Staphylococcus aureus is the most common infecting microorganism. Although any bone can potentially develop osteomyelitis, long-bone, vertebral, and foot osteomyelitis account for the majority of cases. Confirmatory diagnosis of osteomyelitis often depends on the results of a bone biopsy and bone cultures. Radiologic and laboratory studies are often helpful in leading to the diagnosis, determining the extent of the disease, and following up selected patients with osteomyelitis. Optimal therapy for osteomyelitis requires the collaboration of a multidisciplinary team of physicians. Debridement is often needed in contiguous osteomyelitis, whereas acute hematogenous and vertebral osteomyelitis can often be treated with a prolonged course of antimicrobial therapy. PMID- 17127198 TI - Infection and musculoskeletal conditions: Infectious myositis. AB - Infectious myositis, an infection of the skeletal muscle(s), is uncommon. This clinical entity may be caused by viral, bacterial, fungal, and parasitic pathogens. Viral etiologies typically cause diffuse myalgias and/or myositis, whereas bacteria and fungi usually lead to a local myositis which may be associated with sites compromised by trauma or surgery and are more common among immunocompromised patients. Localized collections within the muscles are referred to as pyomyositis. Other pyogenic causes of myositis include gas gangrene, group A streptococcal myonecrosis, and other types of non-clostridial myonecrosis. Early recognition and treatment of these conditions are necessary as they may rapidly become life-threatening. PMID- 17127199 TI - Infection and musculoskeletal conditions: Lyme borreliosis. AB - Lyme borreliosis is a tick-borne infectious disease caused by the spirochaetes Borrelia burgdorferi, B. garinii and B. afzelii. It comprises a wide spectrum of symptoms affecting skin, musculoskeletal system, heart, eyes, central and peripheral nervous system. The diagnosis is based on clinical findings in combination with detection of specific IgM and/or IgG antibodies. Diagnostic problems arise from patients with non-specific symptoms and positive IgG antibody detection. Adequate antibiotic therapy cures more than 90% of the patients. However, in some patients repeated therapy is necessary and a small number of patients develop chronic arthritis or other features. While there is currently no vaccine available, prevention of tick bite is the most effective prophylaxis. PMID- 17127200 TI - Infection and musculoskeletal conditions: Reactive arthritis. AB - Reactive arthritis (ReA) has been recognized as a clinical disease entity for nearly 100 years. The prevalence is estimated to be 30-40/100,000 adults. The HLA B27-associated form is part of the spondyloarthritis concept. According to the current hypothesis the arthritis follows a primary extra-articular infection and is characterized by the presence of bacterial antigen and/or of viable but non culturable bacteria persisting within the joint. Pathogenesis involves the modification of host cells by pathogen-associated molecular patterns (PAMPs, e.g. lipopolysaccharide), bacterial effector proteins, the adaptive immune system, and the genetic background. Up to 30% of patients develop chronic symptoms, and therapeutic options for these patients are still limited. Data for recommendations to apply conventional disease-modifying anti-rheumatic drugs (DMARDs) are rare; however, sulfasalazine seems to be effective, and first reports on agents that block tumour necrosis factor (TNF) are promising. Combination therapy of several antibiotics might open the window to curing the disease; however, controlled clinical studies are needed. PMID- 17127201 TI - Infection and musculoskeletal conditions: Viral causes of arthritis. AB - Several viruses cause postinfectious arthritis. The disease is a typical manifestation of arthritogenic alphaviruses, rubella virus and human parvovirus B19. In addition, arthritis is not uncommon after infection by HIV, cytomegalovirus, hepatitis B virus, hepatitis C virus, or Epstein-Barr virus (EBV). Also prolonged arthritis may result from viral infections, particularly with alphaviruses and human parvovirus B19. Viruses such as EBV and B19 may have significant roles in initiating chronic arthropathies, which in some cases may be indistinguishable from rheumatoid arthritis. PMID- 17127202 TI - Infection and musculoskeletal conditions: Rheumatologic complications of HIV infection. AB - The pandemic caused by the human immunodeficiency virus (HIV) has entered its second quarter-century, with 40 million people now affected worldwide - particularly in Africa, where the impact has been most devastating. A complex array of rheumatic disease manifestations has been described, including diseases specific to HIV infection such as HIV-associated arthritis and the diffuse infiltrative lymphocytosis syndrome; other conditions which occur prominently in HIV-positive individuals include vasculitis, reactive and psoriatic arthritis and HIV-associated polymyositis, opportunistic musculoskeletal infections, and finally disorders that were originally ameliorated by HIV infection, such as rheumatoid arthritis and lupus. Effective antiretroviral treatment ameliorates many of these disorders; however, the introduction of highly active antiretroviral treatment (HAART) has introduced a new spectrum of disorders and new challenges confronting the clinician, including osteonecrosis, rhabdomyolysis, and, with immune reconstitution, the appearance de novo of a variety of autoimmune disorders and phenomena. PMID- 17127203 TI - Infection and musculoskeletal conditions: Bacterial and opportunistic infections during anti-TNF therapy. AB - Tumour necrosis factor alpha (TNF-alpha) plays a crucial role in host defence against bacterial infections. Summarizing the results, the findings of immunological and clinical research suggest a higher infection risk in rheumatoid arthritis and ankylosing spondylitis patients receiving anti-TNF treatment. This is especially true for granulomatous infections in patients treated with the monoclonal TNF-alpha antibodies infliximab or adalimumab. Furthermore, patients treated with TNF inhibitors have a higher susceptibility to infections because of their higher active and more severe disease. Therefore, patients receiving anti TNF treatment should be closely monitored for serious infections. A rapid and sufficient treatment of infections that are not mild and transient is recommended. There are atypical signs and symptoms as well as atypical pathogen that should be considered. Patients should be educated about how to avoid infectious complications. PMID- 17127204 TI - Infection and musculoskeletal conditions: Imaging of musculoskeletal infections. AB - Imaging procedures are routinely used to evaluate patients suspected of having musculoskeletal infection. Radiographs should be performed whenever musculoskeletal infection is suspected. Even when not diagnostic, radiographs are useful. They provide an anatomic overview of the region of interest, including pre-existing conditions that could influence the selection and interpretation of subsequent procedures. Magnetic resonance imaging (MRI) is sensitive, provides superb anatomic detail, does not use ionizing radiation, and is rapidly completed. This technique is especially valuable for septic arthritis, spinal osteomyelitis, and diabetic foot infections. Among the radionuclide procedures, three-phase bone imaging is readily available, and very accurate in unviolated bone. Labeled leukocyte imaging should be used in cases of 'complicating osteomyelitis' such as prosthetic joint infections. This test is also useful in unsuspected diabetic pedal osteomyelitis and the neuropathic joint. Gallium imaging is a useful adjunct to MIR in spinal infection. 18F-2-fluoro-2-deoxy-D glucose positron emission tomography (FDG-PET) will likely play an important role, especially in the evaluation of spinal infection. PMID- 17127205 TI - Who will respond the next time? PMID- 17127206 TI - Disaster nursing book seeking contributors. PMID- 17127207 TI - National disaster life support education for emergency nurses. PMID- 17127208 TI - Challenges and opportunities of nursing care in special-needs shelters. AB - Hurricanes Katrina and Rita forced many individuals along the coast of Texas and Louisiana to seek shelter inland. Among the evacuees were residents with special needs and residents of nursing homes and group homes caring for mentally retarded and physically disabled persons. Many nurses volunteered to provide health care for those in need. This article discusses challenges and opportunities that were encountered by nurses volunteering in special-needs shelters. Issues related to human and physical resources, patient care, and confidentiality are discussed including lessons learned. As nurses who cared for evacuees in the shelter, it is hoped some of the lessons learned can be utilized in future disasters. PMID- 17127209 TI - Better prepared but spread too thin: the impact of emergency preparedness funding on local public health. AB - Local public health authorities (LPHAs) are recognized as playing critical roles in response to biological, chemical, and other health emergencies. An influx of emergency preparedness funding has created new and expanding responsibilities for LPHAs. Concern that funding for emergency response is diverting attention and resources away from other core public health responsibilities is increasing. In order to determine the impact of emergency preparedness funding on public health infrastructure, qualitative interviews with 27 LPHAs in the metro-Boston area were conducted as part of an on-going evaluation of preparedness planning in Massachusetts. Feedback on the benefits and challenges of recent emergency preparedness planning mandates was obtained. Benefits include opportunities to develop relationships within and across public health departments and increases in communication between local and state authorities. Challenges include budget constraints, staffing shortages, and competing public responsibilities. Policy recommendations for improving planning for emergency response at the local level are provided. PMID- 17127210 TI - CCATT: a military model for civilian disaster management. AB - When major disasters incapacitate hospitals and definitive care facilities-as Hurricane Katrina did in 2005-a crisis point is rapidly reached. Critical care services are often the first to be overwhelmed. Personal experiences and regional disaster plans were examined in the wake of Hurricane Katrina to uncover shortfalls in delivery of care and resources. A search was undertaken for a viable model for delivering critical care services in the immediate post-disaster period. Such a model already exists in the US Air Force's (USAF) Critical Care Air Transport Teams (CCATT). These teams have functioned well during recent military conflicts by providing both ground critical care and transport of high risk, severely injured patients. The need for augmented critical care and transport resources in the face of overwhelming casualties in the civilian environment does not require a de novo construct. The USAF's CCATT model should be easily adaptable to the civilian disaster scenario. PMID- 17127211 TI - Functional characterization of gibberellin-regulated genes in rice using microarray system. AB - Gibberellin (GA) is collectively referred to a group of diterpenoid acids, some of which act as plant hormones and are essential for normal plant growth and development. DNA microarray technology has become the standard tool for the parallel quantification of large numbers of messenger RNA transcripts. The power of this approach has been demonstrated in dissecting plant physiology and development, and in unraveling the underlying cellular signaling pathways. To understand the molecular mechanism by which GA regulates the growth and development of plants, with reference to the monocot model plant-rice, it is essential to identify and analyze more genes and their products at the transcription and translation levels that are regulated by GA. With the availability of draft sequences of two major rice types, indica and japonica rice, it has become possible to analyze global expression profiles of genes on a genome scale. In this review, the progress made in finding new genes in rice leaf sheath using microarray system and their characterization is discussed. It is believed that the findings made in this regard have important implications for understanding the mechanism by which GA regulates the growth and development of rice. PMID- 17127212 TI - Glia maturation factor gamma (GMFG): a cytokine-responsive protein during hematopoietic lineage development and its functional genomics analysis. AB - Human hematopoiesis was evaluated using the techniques of controlled stem cell differentiation, two-dimensional gel electrophoresis-based proteomics, and functional genomics. We provide the first report that glia maturation factor gamma (GMFG) is a cytokine-responsive protein in erythropoietin-induced and granulocyte-colony stimulating factor-induced hematopoietic lineage development. Results from global functional genomics analysis indicate that GMFG possesses several other features: hematopoietic tissue-specific gene expression, a promoter concentrated with high-score hematopoiesis-specific transcription factors, and possible molecular coevolution with a rudimentary blood/immune system. On the basis of our findings, we hypothesize that GMFG is a hematopoietic-specific protein that may mediate the pluripotentiality and lineage commitment of human hematopoietic stem cells. PMID- 17127213 TI - A combined computational and experimental study on the structure-regulation relationships of putative mammalian DNA replication initiator GINS. AB - GINS, a heterotetramer of SLD5, PSF1, PSF2, and PSF3 proteins, is an emerging chromatin factor recognized to be involved in the initiation and elongation step of DNA replication. Although the yeast and Xenopus GINS genes are well documented, their orthologous genes in higher eukaryotes are not fully characterized. In this study, we report the genomic structure and transcriptional regulation of mammalian GINS genes. Serum stimulation increased the GINS mRNA levels in human cells. Reporter gene assay using putative GINS promoter sequences revealed that the expression of mammalian GINS is regulated by 17beta-Estradiol stimulated estrogen receptor alpha, and human PSF3 acts as a gene responsive to transcription factor E2F1. The goal of this study is to present the current data so as to encourage further work in the field of GINS gene regulation and functions in mammalian cells. PMID- 17127214 TI - Comparative proteome analysis of breast cancer and adjacent normal breast tissues in human. AB - Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI TOF/TOF-MS), incorporated with online database searching, were performed to investigate differential proteins of breast cancer and adjacent normal breast tissues. Considering that serum albumin is abundantly presented in normal control samples, 15 differential spots detected in 11 out of 12 (91.7%) breast cancer samples were identified by online SIENA-2DPAGE database searching and MALDI TOF/TOF-MS analysis. The results indicate that pathological changes of breast cancer are concerned with augmentation of substance metabolism, promotion of proteolytic activity, decline of activity of some inhibitors of enzymes, and so on. Some important proteins involved in the pathological process of breast cancer with changed expression may be useful biomarkers, such as alpha-1-antitrypsin, EF 1-beta, cathepsin D, TCTP, SMT3A, RPS12, and PSMA1, among which SMT3A, RPS12, and PSMA1 were first reported for breast cancer in this study. PMID- 17127215 TI - Evaluation of six methods for estimating synonymous and nonsynonymous substitution rates. AB - Methods for estimating synonymous and nonsynonymous substitution rates among protein-coding sequences adopt different mutation (substitution) models with subtle yet significant differences, which lead to different estimates of evolutionary information. Little attention has been devoted to the comparison of methods for obtaining reliable estimates since the amount of sequence variations within targeted datasets is always unpredictable. To our knowledge, there is little information available in literature about evaluation of these different methods. In this study, we compared six widely used methods and provided with evaluation results using simulated sequences. The results indicate that incorporating sequence features (such as transition/transversion bias and nucleotide/codon frequency bias) into methods could yield better performance. We recommend that conclusions related to or derived from Ka and Ks analyses should not be readily drawn only according to results from one method. PMID- 17127216 TI - Phylogenetic analysis of RhoGAP domain-containing proteins. AB - Proteins containing an Rho GTPase-activating protein (RhoGAP) domain work as molecular switches involved in the regulation of diverse cellular functions. The ability of these GTPases to regulate a wide number of cellular processes conies from their interactions with multiple effectors and inhibitors, including the RhoGAP family, which stimulates their intrinsic GTPase activity. Here, a phylogenetic approach was applied to study the evolutionary relationship among 59 RhoGAP domain-containing proteins. The sequences were aligned by their RhoGAP domains and the phylogenetic hypotheses were generated using Maximum Parsimony and Bayesian analyses. The character tracing of two traits, GTPase activity and presence of other domains, indicated a significant phylogenetic signal for both of them. PMID- 17127217 TI - Comparative analysis of transcription start sites using mutual information. AB - The transcription start site (TSS) region shows greater variability compared with other promoter elements. We are interested to search for its variability by using information content as a measure. We note in this study that the variability is significant in the block of 5 nucleotides (nt) surrounding the TSS region compared with the block of 15 nt. This suggests that the actual region that may be involved is in the range of 5-10 nt in size. For Escherichia coli, we note that the information content from dinucleotide substitution matrices clearly shows a better discrimination, suggesting the presence of some correlations. However, for human this effect is much less, and for mouse it is practically absent. We can conclude that the presence of short-range correlations within the TSS region is species-dependent and is not universal. We further observe that there are other variable regions in the mitochondrial control element apart from TSS. It is also noted that effective comparisons can only be made on blocks, while single nucleotide comparisons do not give us any detectable signals. PMID- 17127218 TI - Optimal allocation of replicates for measurement evaluation studies. AB - Optimal experimental design is important for the efficient use of modern high throughput technologies such as microarrays and proteomics. Multiple factors including the reliability of measurement system, which itself must be estimated from prior experimental work, could influence design decisions. In this study, we describe how the optimal number of replicate measures (technical replicates) for each biological sample (biological replicate) can be determined. Different allocations of biological and technical replicates were evaluated by minimizing the variance of the ratio of technical variance (measurement error) to the total variance (sum of sampling error and measurement error). We demonstrate that if the number of biological replicates and the number of technical replicates per biological sample are variable, while the total number of available measures is fixed, then the optimal allocation of replicates for measurement evaluation experiments requires two technical replicates for each biological replicate. Therefore, it is recommended to use two technical replicates for each biological replicate if the goal is to evaluate the reproducibility of measurements. PMID- 17127219 TI - Experimental osteomyelitis: what have we learned from animal studies about the systemic treatment of osteomyelitis? AB - Clinical trials of systemic antibiotic treatment of osteomyelitis are difficult to perform for many reasons, such as low incidence rate of osteomyelitis, variety of anatomic locations, stage and etiologic agents. In this article, we reviewed the experimental studies on osteomyelitis available in the English medical literature since 1968, to ascertain their actual and potential impact on the treatment of human osteomyelitis. Major results are summarized and topics of major interest, such as reproducibility of animal models, predictive value of animal models, correlation of pharmacokinetics between different animals and humans, and the correlation of outcome between animal and clinical studies are discussed. Most of the reviewed animal models are reproducible and dependable. However, establishing the right dose regimen in animals appeared a critical factor, which might undermine the predictive value of the experimental study. Due to difficulties in comparing results of animal and human studies, the predictive value of animal studies about osteomyelitis is still unclear. However, animal models gave valuable information to the clinician for choosing the minimal duration of antibiotic treatment. Even though the use of antibiotic combinations was associated with better outcome in the majority of animal studies, such a finding seems to have limited impact on clinical practice. PMID- 17127220 TI - Microbiological and epidemiological characterization of imipenem-resistant Pseudomonas aeruginosa strains from a Brazilian tertiary hospital: report from the SENTRY Antimicrobial Surveillance Program. AB - OBJECTIVES: To evaluate the antimicrobial susceptibility profile, the genetic similarity, and the mechanisms of carbapenem resistance among imipenem-resistant Pseudomonas aeruginosa isolates collected from a Brazilian tertiary teaching hospital. METHODS: Seventy-eight consecutive samples of P. aeruginosa were evaluated during 2000 and 2001. The antimicrobial susceptibility was evaluated by reference broth microdilution methods and the imipenem-resistant isolates were screened for metallo-beta-lactamase (MbetaL) production throughout disc approximation test and MbetaL Etest strips and isolates with positive screen test result were submitted to PCR assays using primers blaIMP-1, bla VIM-1, blaVIM-2 e blaSPM-1. The genetic similarity of MbetaL-producing strains was evaluated by automated ribotyping for epidemiological typing purpose. RESULTS: Resistance rates were high to the majority of antimicrobial agents tested except polymyxin B, which inhibited all samples at the Clinical and Laboratory Standards Institute breakpoint (< or = 2 microg/ml). Twenty-nine (37.2%) isolates were resistant to imipenem and these isolates showed great genomic variability. MbetaL production was demonstrated in two imipenem-resistant isolates, which were detected using blaSPM-1 and blaIMP-2-specific primers. Sequence analysis revealed the presence of blaSPM-1 and a novel blaIMP-type gene, blaIMP-16. CONCLUSION: The results of this study showed high resistance rates to the majority of antimicrobial agents among P. aeruginosa samples. High imipenem resistance rates were probably due to continuous selection of resistant mutants. The production of MbetaL did not represent a frequent mechanism of carbapenem resistance in this medical center; but a novel MbetaL was identified. Continued antimicrobial surveillance and infection control measures should be emphasized to minimize the emergence and dissemination of antimicrobial resistance. PMID- 17127221 TI - Characterization of antimicrobial resistance and class 1 integrons found in Escherichia coli isolates from human stools and drinking water sources in Jordan. AB - This study demonstrates that Escherichia coli isolates from human stools showed mostly higher minimum inhibitory concentrations (MICs) and significant rates of resistance (32%-67%, P<0.05) than Escherichia coli water isolates in Jordan, as follows: ampicillin (67% vs 28%), trimethoprim/sulfamethoxazole (67% vs 28%) nalidixic acid (63% vs 20%), cefuroxime (32% vs 4%), gentamicin (32% vs 17%), norfloxacin (32% vs 12%) and tetracycline (33% vs 16%). The prevalence of integron integrase genes (Intl1) in these isolates was also significantly higher in patients' stools (67%, P <0.05) than in water (36%), but the distribution of Sul 1/Sul 2 or both in association with postive Intl1 and resistance to ampicillin and sulfamethoxazole was not significantly higher (74% versus 62%, P <0.05) in isolates from stool and water. Plasmid profiles of representative multiresistant E. coli isolates from both sources indicated the presence of two common plasmids (49,25 kb) in 11/12 (91.6%), and all E. coli transconjugants were positive for class 1 integron markers (Intl 1, Sul 1 and Sul2) and mostly associated with three transferable drug-resistant determinants to ampicillin, sulfamethoxazole and tetracycline. These results indicate that class 1 integrons with conjugative R-plasmids are common and transferable among commensal antimicrobial multiresistant E. coli isolated from human feces and drinking water sources in Jordan. PMID- 17127222 TI - In vitro studies on the release of isoniazid incorporated in poly(epsilon caprolactone). AB - A polymeric micro- and nanosphere formulation using poly (epsilon-caprolactone) (PCL) to entrap an antituberculosis drug, isoniazid (INH), was developed and characterized. The microspheres were prepared by a solvent evaporation method using ethyl acetate, PCL and INH as the organic phase and water and Tween 40 as the aqueous phase. The nanospheres were prepared by a spontaneous emulsification solvent diffusion method using 40% ethanol in acetone (v/v), PCL and INH as the organic phase and water and Tween 40 as the aqueous phase. After freeze-drying, these systems were characterized by scanning electron microscopy (SEM), particle size analysis, determination of entrapped INH content, in vitro INH release and brine shrimp toxicity bioassay. PMID- 17127223 TI - Prevalence of macrolide resistance genes among staphylococci in Cyprus. AB - The aims of the present study were to evaluate the frequency of macrolide resistant staphylococci in Cyprus and to examine the phenotypic and genotypic characteristics of these isolates. Antimicrobial susceptibility testing was performed by broth microdilution method and the macrolide resistance determinants were detected by PCR. The relatedness among the isolates was examined by pulsed field gel electrophoresis. Ninety-six (67.61%) of the 142 Staphylococcus aureus and 19 (59.4%) of the 32 coagulase-negative staphylococci were resistant to erythromycin. Among the 115 erythromycin-resistant staphylococci, 70 expressed the MLSB-inducible phenotype, 38 the MLSB-constitutive, and 7 the MS. The predominant genes associated with macrolide resistance were the ermA for S. aureus and the ermC for coagulase-negative staphylococci, detected in 90.62% and 47.37% of the isolates respectively. Dissemination of one clone carrying the ermA gene accounted for macrolide resistance in the majority of S. aureus isolates. PMID- 17127224 TI - Serum and prostatic tissue concentrations of moxifloxacin in patients undergoing transurethral resection of the prostate. AB - The spectrum of pathogens causing chronic bacterial prostatitis comprises Gram negative, Gram-positive and atypical microorganisms. Because of its broad spectrum of activity, the group 4 fluoroquinolone moxifloxacin might be a suitable antibiotic for treatment of bacterial prostatitis. The aim of this prospective study was to investigate the penetration of moxifloxacin into prostatic tissue in patients with benign prostatic hyperplasia. Patients received a single dose of moxifloxacin 400 mg in an 1 hour lasting infusion (250 ml) for perioperative prophylaxis before undergoing transurethral resection of the prostate (TURP). Serum concentrations were determined in all patients before infusion, at the end of infusion (time point 0), 0.5, 1 and 2 h after the end of infusion. Patients were randomized for tissue sampling either 0, 0.5, 1 or 2 h after the end of infusion. At beginning of TURP approximately 1 g of tissue was sampled for analysis. Concentrations of moxifloxacin in serum and tissue were determined by HPLC. 39 patients were evaluated. Median serum and prostatic tissue concentrations peaked at 0 h (4.94 mg/ L and 8.50 mg/ kg, respectively). The lowest concentrations were quantified at 2 h after the end of infusion (2.46 mg/ L and 3.88 mg/ kg, respectively). The prostatic tissue concentrations of moxifloxacin were approximately twice as high as in corresponding serum. At the end of infusion the tissue and serum concentrations seemed to be already equilibrated, as their ratios did not differ significantly during the time of investigation. After an intravenous infusion of 400 mg the serum and prostatic tissue concentrations of moxifloxacin were well above the MIC values of most important prostatic pathogens. The high tissue/ serum ratio and the extended antibacterial spectrum suggests active concentration in the prostate which may translate into increased efficacy compared to group 2 and 3 fluoroquinolones in the treatment of chronic bacterial prostatitis. PMID- 17127225 TI - Appropriateness of empiric gentamicin and vancomycin therapy for bacteremias in chronic dialysis outpatient units in the era of antibiotic resistance. AB - Bacteremias in inpatient chronic HD units have been described, but there is little information on bacteremias in ambulatory HD units. To determine the frequency of bacteremia and pathogen distribution in ambulatory chronic HD units, we retrospectively reviewed our experience with 107 bacteremias in 5 chronic ambulatory HD units over a 3 year period. The object of the study was twofold. The first objective was to determine if bacteremias in ambulatory HD setting were substantially different in frequency or type than in the inpatient HD setting. Secondly, febrile patients suspected of having bacteremia in chronic HD patients are often empirically treated with vancomycin and gentamicin. Chronic HD patients require repeated and frequent venous access for HD. Bacteremias are common in chronic HD patients and may be primary or secondary and are often related to venous access site infections. The distributions of bacteremia pathogens in chronic HD patients are predominantly reflective of skin flora, i.e., staphylococci and to lesser extent aerobic Gram-negative bacilli. After S. aureus (MSRA/MSSA) and coagulase-negative staphylococcus (CoNS), enterococci are the next most important Gram-positive pathogens in bacteremic HD patients. Most strains of E. faecalis are sensitive to vancomycin and for practical purposes should be considered as vancomycin sensitive enterococci (VSE). In contrast, most strains of E. faecium are resistant to vancomycin and should be considered as vancomycin resistant enterococci (VRE). We retrospectively reviewed 107 patients on chronic ambulatory HD to determine the adequacy of empiric vancomycin and gentamicin prophylaxis. We found amikacin is preferred to gentamicin and that meropenem is an effective alternate substitution for gentamicin and vancomycin combination therapy. PMID- 17127226 TI - A pilot study on prevention of catheter-related urinary tract infections with fluoroquinolones. AB - The objective of this multicenter, randomized, controlled, parallel group trial was to evaluate the efficacy of levofloxacin 250 mg oral, once daily (LVFX), placebo one tablet oral once daily (Placebo [P] group) and ciprofloxacin (CPFX) 500 mg oral, twice daily (single blind), prophylaxis in preventing bacteriuria (> or = 10(3) CFU/ml) in post-surgical catheterized patients. In the modified intention-to-treat (M-ITT) population of the 82 enrolled patients, negative bacteriuria was observed in 92% of LVFX group, in 80% of P group and in 100% of CPFX group while in the per-protocol (PP) population figures were: 100%, 86.4% and 100% respectively. Only one symptomatic urinary tract infection and one surgical wound infection were observed in the P group. Both drugs were well tolerated, showing a safety profile comparable to placebo. The high frequency of negative bacteriuria in the placebo group sounds encouraging as it underlines that the adoption of closed urinary drainage system catheters in hospital setting may reduce the frequency of hospital-acquired infections. PMID- 17127227 TI - Impact of carbapenem administration on systemic endotoxemia in patients with severe sepsis and Gram-negative bacteremia. AB - In order to investigate the effect of carbapenems on systemic endotoxemia, 20 patients with severe sepsis due to ventilator-associated pneumonia and Gram negative bacteremia were enrolled; 10 (group A) were administered 1 g t.i.d. of imipenem/cilastatin and 10 (group B) 2 g t.i.d. of meropenem. Blood was sampled at 0 time and after 1, 2, 4, 6, 12, 24, 36, 48, 60, 72, 84 and 96 hours for detection of endotoxins (LPS), interleukin-6 (IL-6), C-reactive protein (CRP) and drug levels. LPS were determined by the QCL-1000 LAL assay, IL-6 by an enzymeimmunoassay, CRP by nephelometry and carbapenem levels by a microbiological assay. We did not find that carbapenems had any effect on the kinetics of LPS and CRP; IL-6 of group A was lower than group B at 72 and 84 hours. No correlation was observed between drug levels of any carbapenem and LPS, IL-6 or CRP. It is concluded that in septic patients with Gram-negative bacteremia administration of either imipenem or meropenem did not affect systemic endotoxemia. The above data support the safe administration of both carbapenems in patients with severe sepsis. PMID- 17127228 TI - Efficacy of N-acetyl-cysteine in combination with thiamphenicol in sequential (intramuscular/aerosol) therapy of upper respiratory tract infections even when sustained by bacterial biofilms. AB - A total of 102 patients with recurrent upper respiratory tract infections underwent microbiological exploration with appropriate sampling and direct biopsies of the infected sites. Therapy was then started and on day 1 each patient received two intramuscular injections of thiamphenicol glycinate acetylcysteinate (TGA). From day 2 to 10 sequential therapy with the same drug was continued employing TGA administered by aerosol. All putative etiologic agents recovered were susceptible to thiamphenicol and only 24 demonstrated the ability to produce in vitro biofilms. The organisms comprised 10 Staphylococcus aureus, 6 Streptococcus pyogenes, 4 Streptococcus pneumoniae and 3 Haemophilus influenzae. Of the 24 subjects in whom biofilms were demonstrated to be present in vivo by Scanning Electron Microscopy, clinical and bacteriological cure was obtained in 21 cases (87.5%) following sequential therapy with TGA. Failures were considered to be persistent signs and symptoms at day 15 after initiation of treatment and lack of eradication of 3 S. aureus strains, despite their in vitro susceptibility to thiamphenicol. Very few adverse events attributable to TGA were reported in this cohort of patients. In no case was discontinuation of treatment deemed necessary by the attending physician. PMID- 17127229 TI - Expression of genes related to activity of oxaliplatin and 5-fluorouracil in endoscopic biopsies of primary esophageal cancer in patients receiving oxaliplatin, 5-flourouracil and radiation: characterization and exploratory analysis with survival. AB - With a goal of identifying relations between gene expression and response (mucosal or pathological) or survival in esophageal cancer patients (stages II to IV) receiving oxaliplatin, 5-fluorouracil (5FU) and radiation, we measured in endoscopic primary tumor biopsies from 38 patients, the expression of seven genes (gammaGCS, gammaGT, MRP-2, ERCC-1, XPA, TS and DPD) prior to treatment, 1 week following oxaliplatin alone and at the end of the combined radio-chemotherapy cycle using real time QRT-PCR. A higher pretreatment level of XPA was related to shorter survival with a hazard ratio of 2.43 (90% confidence interval 1.09 to 5.43) using Cox regression modeling. However, multivariate analysis with a Cox model indicated low expression of XPA or TS and combined stages II and III had a higher probability of survival (for XPA: hazard ratio 3.0 and 90% C.I. of 1.3 to 6.9, with adjustment for stage included; for TS: hazard ratio is 1.98 with 90% C.I. of 0.94 to 4.20. The expression of TS, gammaGCS, ERCC-1 and MRP-2 declined from D 1 to the end of the cycle (p<0.05, sign test). A validation and further understanding of the findings need to be carried out in a larger study with a more homogeneous population of patients. PMID- 17127230 TI - Downregulation of xIAP expression by small interfering RNA inhibits cellular viability and increases chemosensitivity to methotrexate in human hepatoma cell line HepG2. AB - The aim of this study was to investigate whether downregulating the expression of xIAP by RNAi (RNA interference) technology can induce the apoptosis of HepG2 cells, inhibit cellular viability and increase chemosensitivity of cancer cells. HepG2 cells were transfected with U6 promoter plasmids coding for short interfering RNAs (siRNAs) targeting xIAP. RT-PCR and western blot analysis were used to assess the mRNA and protein levels of xIAP expression. T he suppression efficiency o f xIAPby RNAi was evaluated using the MTT assay for cellular viability and Annexin V-PI binding assay for apoptosis. These results showed that siRNAs reduced cellular viability and increased cellular apoptosis. Moreover, downregulation of xIAP expression enhanced the chemosensitivity of HepG2 cells to methotrexate. These results suggest that the downregulation of xIAP by RNAi could potentially be used as a therapeutic strategy for human hepatocellular carcinoma. PMID- 17127231 TI - Cetuximab in the treatment of metastatic colorectal cancer: a model-based cost effectiveness analysis. AB - Cetuximab (Erbitux) has shown activity in patients with metastatic colorectal cancer (mCRC). To evaluate the cost-effectiveness of this drug combined with irinotecan in mCRC, a model-based cost-effectiveness analysis (CEA) was performed. Data on cetuximab obtained from Medline in December 2004 and from the 2004 ASCO-meeting were analyzed for life years gained (LYG) with regard to the use of this monoclonal antibody (MAb). Norwegian prices as of January 2005 were employed. The LYG ranged between 1.7 and 2.0 years. The median cost per patient treated was calculated to 34,256 Euro to 45,764 Euro yielding a cost per LYG in the range between 205,536 Euro and 323,040 Euro. Sensitivity analysis documented price of cetuximab and survival gain to be the major factors influencing the cost effectiveness ratio. In conclusion, the analysis indicates cetuximab to be a promising, but very expensive antibody. PMID- 17127232 TI - A prospective randomized study of irinotecan (CPT-11), leucovorin (LV) and 5 fluorouracil (5FU) versus leucovorin and 5-fluorouracil in patients with advanced colorectal carcinoma. AB - The purpose of this study was to compare the activity and toxicity of an irinotecan (CPT-11), leucovorin (LV) and 5-fluorouracil (5FU) combination with a standard regimen of 5FU and LV, in patients with advanced colorectal carcinoma. One hundred and sixty patients were randomized; 80 patients (group A) received LV 20 mg/m(2) bolus i.v. and 5FU 425 mg/m(2) bolus i.v. on days 1-5, every 28 days; 80 patients (group B) received CPT-11 80 mg/m(2) (30-90 min i.v. infusion), followed by LV 20 mg/m(2) bolus i.v. and 5FU 425 mg/m(2) bolus i.v. on days 1, 8, 15, 22, 29, and 36, every 8 weeks. The overall response rate was 30% and 47.5% in groups A and B respectively. Progression-free survival was significantly higher in the triple-drug combination arm (median 7.5 vs. 4.5 months; p= 0. 0335). However, overall survival did not differ significantly between the two arms (15 months vs. 14 months for the groups B and A respectively; p=0.3531). The main grade 3 adverse events were diarrhea (19%, in group A vs. 35% in group B; p=0.032) and mucositis (2% vs. 14%; p=0.017). The regimen containing irinotecan showed activity in advanced colorectal cancer. The overall safety data confirm this combination as a well-tolerated treatment. PMID- 17127233 TI - Chemotherapeutic management of small bowel adenocarcinoma associated with Crohn's disease. AB - Four patients with metastatic primary small bowel adenocarcinoma associated with Crohn's disease were successfully treated with low dose combination chemotherapy consisting of 5-fluorouracil, leucovorin and irinotecan with or without gemcitabine. Benefits included prolonged survival, objective responses, response of resistant tumors, downstaging, and a successful secondary complete resection (Ro) with a durable remission. PMID- 17127234 TI - Clinical efficacy and tolerability of caspofungin in a renal transplant patient with Aspergillus flavus lung infection: case report. AB - Organ transplant recipients are at increased risk for severe invasive aspergillosis, and amphotericin deoxycholate has been the standard treatment for many years. Currently, however, lipid formulations are preferred due to their few side effects. Also, a number of new antifungal drugs have been developed including new azoles and echinocandins. Caspofungin is the first of the echinocandin derivatives patented to treat patients with invasive aspergillosis who are refractory or intolerant to other therapies. A renal transplant patient on immunosuppressive treatment with chronic hepatitis B virus infection was admitted with fever, hemophthisis and lung consolidation, diagnosed to be probably caused by Aspergillus flavus. The patient developed cholestatic hepatitis most likely related to itraconazole. Clinical failure and in vitro itraconazole resistance of the isolate was also documented while the patient was receiving itraconazole at a reduced dosage. Caspofungin was administered once a day as ambulatory treatment and was well tolerated. Clinical improvement was observed after 6 weeks of treatment and no hepatic toxicity was documented. Caspofungin seems to be a potentially useful antifungal agent in renal transplant patients with invasive aspergillosis. Further evaluation of the efficacy of caspofungin is needed. PMID- 17127235 TI - Post-neurosurgical meningitis due to multidrug-resistant Acinetobacter baumanii treated with intrathecal colistin: case report and review of the literature. AB - Intrathecal colistin (Polymxin E) is becoming an important option for the treatment of post-neurosurgical meningitis caused by multidrug resistant (MDR) Acinetobacter baumannii. We report a case of 28-year-old man who developed meningitis due to MDR A. baumannii associated with an external ventricular drain. The patient was cured using a 4-week course of intrathecal colistin 3.2 mg via external ventricular drain (EVD) daily without any serious side effects. PMID- 17127236 TI - Severe Legionnaires' disease successfully treated with levofloxacin and azithromycin. PMID- 17127237 TI - Lack of usefulness of the immunochromatographic urinary pneumococcal antigen test in maximizing pneumococcal yield in a community-acquired pneumonia clinical trial. PMID- 17127238 TI - Antimicrobial resistance of respiratory pathogens isolated from pediatric patients. PMID- 17127239 TI - Plasma insulin levels predict the development of atherosclerosis when IRS2 deficiency is combined with severe hypercholesterolemia in apolipoprotein E-null mice. AB - Atherosclerosis is increased in type 2 diabetic patients but the precise mechanisms underlying this predisposition remain vague. Mice deficient for insulin receptor substrate 2 (IRS2) develop type 2-like diabetes and thus, provide a model to explore the molecular connection between deranged carbohydrate metabolism and atherosclerosis. To explore the relationship between defective insulin signalling and atherosclerosis, we have examined the development of atherosclerosis in the following groups of fat-fed mice: wild-type, diabetic Irs2 null (Irs2-/-), atherosclerosis-prone apolipoprotein E-null (apoE-/-), and doubly deficient apoE-/- Irs2-/-. Surprisingly, glucose levels of apoE-/- Irs2-/- mice were comparable to those seen in wild-type and apoE-/- and significantly lower than in Irs2-/- mice. Irs2-/- and apoE-/- Irs2-/- were hyperinsulinemic compared to wild-type and apoE-/- mice. Atherosclerotic lesions were barely detectable in wild-type and Irs2-/- mice, which displayed moderate hypercholesterolemia (approximately 280 mg/dL). Notably, atherosclerosis was significantly enhanced in apoE-/- Irs2-/- compared with apoE-/- mice, although both models displayed similar levels of severe hypercholesterolemia (>600 mg/dL). Circulating insulin levels predicted atherosclerotic lesion burden in apoE-/- Irs2-/- mice. Our results suggest that hyperinsulinemia as a result of Irs2 genetic ablation contributes to increased atherosclerosis when combined with severe hypercholesterolemia in the absence of hyperglycaemia (apoE-/- Irs2-/- mice), thus implicating IRS2 as an important modulator of murine hypercholesterolemia dependent atherosclerosis. Future studies are necessary to determine whether IRS2 dysfunction may promote atherosclerosis in normoglycemic, pre-diabetic patients with clinical manifestations of hyperinsulinemia and insulin resistance. PMID- 17127240 TI - Cytochrome P450 expression (CYP) in non-small cell lung cancer. AB - The cytochrome P450 (CYP) is associated with tumor development and progression as well as activation of anti-cancer prodrugs and their metabolic clearance. In this study, we investigated the expression of aryl-hydrocarbon receptor (AH-R) and four CYPs (CYP1A1, CYP2A6, CYP2E1 and CYP3A) as putative diagnostic markers in 78 non-small cell lung cancers (NSCLC) along with clinical features of the patients. In non-small cell lung cancer, the expression of the five markers was mainly observed in adenocarcinoma but not in the most squamous cell cancers. The expression of them in adenocarcinoma was more frequent in females than in males, suggesting that a higher risk of women for developing lung adenocarcinoma might be associated with the frequent expression of AH-R and the CYPs. These factors were also more frequently expressed in early stage adenocarcinoma and more differentiated adenocarcinoma. Multiple types of CYPs are more frequently expressed in early stage of adenocarcinoma than in advanced stage of adenocarcinoma. There were positive relationships among AH-R, CYP1A1, CYP2E1 and CYP3A expressions in adenocarcinoma, which suggests a metabolite-mediated cross talk in the gene regulation of these markers. However, any of them was unrelated with the expression of CYP2A6, suggesting that the gene regulation of CYP2A6 in adenocarcinoma may be different from the other three CYPs. The expression frequency of CYP1A1 and CYP2E1 in tumors is independent of their genetic polymorphism. The survival of the patients with advanced adenocarcinoma expressing more than one of CYPs was lower rate than the patients with those expressing no CYPs, suggesting that the expression of the CYPs in advanced adenocarcinoma may be associated with poor survival. Our results suggest that AH R and 4 CYPs may be good markers for the determination of quality of lung cancer. The information could be useful for the better management of lung cancer by molecular targeting therapy and selection of anti-cancer drug based on individual spectrum of the marker proteins. Therefore, the spectrum of CYP proteins in lung cancer could be useful for changing the present "order-made" therapy to the "tailor-made" therapy. PMID- 17127241 TI - Green tea polyphenols in the prevention of colon cancer. AB - Several plant-based nutrients and non-nutrients that can inhibit mutagenesis and proliferation have been identified. Some of the most promising nutrients identified as chemopreventive agents in colon cancer prevention include isoflavones, curcumin, calcium, vitamin D and more recently Green tea polyphenols (GTP). In addition to inhibiting mutagenesis and proliferation, these compounds are relatively non-toxic, are of low cost and can be taken orally or as a part of the daily diet. Epidemiological and laboratory studies have identified epigallocatechin gallate (EGCG) in green tea polyphenols (GTP), as the most potent chemopreventive agent that can induce apoptosis, suppress the formation and growth of human cancers including colorectal cancers (CRC). It is only logical then, that future clinical studies should focus on examining the efficacy of phytochemicals such as EGCG in cancer chemoprevention as an alternative to pharmacological agents, especially in populations where administration of COX-2 inhibitors, Aspirin and NSAIDS is contraindicated. The goal of this review is to provide the rationale, and discuss the use of EGCG in GTP as a chemopreventive agent for prevention of colon carcinogenesis and present evidence for the efficacy and safety of these agents based on epidemiological, animal, in vitro studies and Phase I clinical trials. PMID- 17127242 TI - MicroRNA: past and present. AB - MicroRNAs (miRNAs) are approximately 22 nucleotide (nt) non-coding RNAs that participate in gene regulation. MiRNAs confer their regulation at a post transcriptional level, where they either cleave or repress translation of mRNAs. Over 3000 identified mature miRNAs exist in species ranging from plants to humans, suggesting that they are ancient players in gene regulation. A relatively small number of miRNAs have been experimentally tested for their function. Of those tested, functions including cell differentiation, proliferation, apoptosis, anti-viral defense and cancer have been proposed. Improved software programs are now able to predict the targets of miRNAs in a more efficient manner, thus facilitating the elucidation of miRNA function. Furthermore, methods such as real time PCR and microarray have been enhanced for studying miRNA expression. Using these tools, scientists are able to discover novel functions for miRNAs. It is possible that miRNAs will be revealed as having a role in virtually every aspect of gene regulation. This review guides readers through the biogenesis of miRNAs, their mechanism of action on their target mRNAs, the functional outcomes of their action on mRNAs and the current techniques to investigate these processes. PMID- 17127243 TI - T cell immune responses to HIV-1. AB - The recent use of multiparametric flow cytometry to monitor T cell immune responses complements traditional assays, such as IFN-gamma ELISPOT, to provide more information on the functional complexity of CD4+ and CD8+ T cell immune responses induced either by natural infection, or by immunization. In this review, we provide a general background on T cell subsets, and describe the cellular immune response during natural HIV-1 infection. We then review T cell responses to current candidate HIV-1 vaccines. Taken together, this helps to formulate our understanding of the immune correlates of protection required for an effective prophylactic HIV-1 vaccine. Finally, we emphasize current dendritic cell based vaccine strategies designed to modulate immunity to establish immune protection against HIV-1. PMID- 17127244 TI - Genetic approaches to complications of prematurity. AB - Over the last 15 years neonatal morbidity and mortality have changed little for very low birth weight (VLBW) babies despite significant technological and therapeutic advances. While clinical trials and animal models have until recently improved outcomes in this gestational age group, further productivity from these traditional sources are not likely. A recent study of monozygotic and dizygotic twins shows that the main determinants of neonatal morbidity and mortality in VLBW babies--bronchopulmonary dysplasia, necrotizing enterocolitis, and intraventricular hemorrhage--have significant genetic components. Incremental improvements in the future, therefore, will likely depend on identification of these genetic components for targeting specific therapies. Cost-effective methods and resources, fueled by the Human Genome and HapMap Projects and recent successes in identifying genes for a small number of complex genetic diseases, are available now and through creative planning and timely implementation would likely yield useful results. PMID- 17127245 TI - Electrogenerated chemiluminescence of CdSe quantum dots dispersed in aqueous solution. AB - Electrogenerated chemiluminescence (ECL) of CdSe quantum dots (QDs) dispersed in aqueous solution was studied with bare electrode. The ECL emission was observed at -1.4 V vs. Ag/AgCl and the ECL spectrum peak is similar to that of the defect photoluminescence (PL) spectrum, indicating the surface defects played a critical role in the emission process. The experiment results suggested that dissolved oxygen had a great effect on the ECL intensity. Other influence factors including the electrochemical parameters and QDs concentration were investigated in detail. As an application of the CdSe QDs ECL, the pyrogallol (1, 2, 3-trihydroxybenzene) was detected in aqueous solution. Under the optimal conditions, a linear relationship between ECL intensity and pyrogallol concentration was obtained in the range from 4.0 x 10(-7) to 2.0 x 10(-5) M with a correlation coefficient of 0.9904 and the limit of detection was 6.6 x 10(-8) M (S/N = 3). A possible mechanism about ECL of QDs was also discussed. PMID- 17127246 TI - Yeast as a model system to study glucose-mediated signalling and response. AB - Glucose is the principal carbon and energy source for a wide variety of cells, ranging from unicellular microorganisms to higher eukaryotic cells. It is taken up by these cells and metabolized to obtain the energy necessary for cell viability. In addition, the presence of this sugar is able to adjust cellular metabolism, regulate gene expression and even influence cell growth. For this reason, glucose is considered as a "hormone". Specifically, it can trigger different signalling pathways that allow cells to adjust their gene expression programmes in response to glucose availability. Elucidating the molecular mechanisms of glucose response in eukaryotes has been greatly aided by studies conducted in the yeast Saccharomyces cerevisiae. This yeast shares with complex multicellular eukaryotes many of the signal transduction components that detect glucose, transmit the corresponding signals to the interior of the cell and make the needed adjustments to cellular metabolism and gene expression. In this manuscript, I will review the current knowledge of some aspects of glucose mediated signalling in yeast and discuss how these results have contributed to the understanding of similar processes in mammalian cells. PMID- 17127247 TI - Genetic aspects of preeclampsia. AB - Preeclampsia has a familial component suggesting that one or more common alleles may act as susceptibility genes. Some families may have "private" predisposing mutations. The central role of the placenta in the pathogenesis of preeclampsia implies that fetal genes contribute to the disease process. Twin studies support the role of maternal and fetal gene interaction. Candidate gene studies have not yielded consistent results. Genome-wide linkage studies provide powerful means to study disease susceptibility genes, and several loci have been mapped. Parent-of origin effect of the STOX1 gene has been suggested on chromosome 10q22 locus in the Dutch population. Maternally inherited missense mutations in the STOX1 gene of the fetus have been shown to co-segregate with the maternal preeclamptic phenotype. Up-regulation of placental leptin expression has been found in several studies and might be of importance in the pathogenesis of preeclampsia. DNA microarray is ideal tool for screening gene expression in preeclamptic tissues, but critical attitude is needed when interpreting the results. The placental DNA and mRNA in maternal plasma hold great promise as novel biomarkers for the prediction of preeclampsia. Finding genes prediposing to preeclampsia will enhance our understanding of the disease mechanism, and might allow identification of prognostic and therapeutic subgroups. PMID- 17127248 TI - Endothelial progenitor cells and preeclampsia. AB - The maternal cardiovascular adaptation to pregnancy involves a complex physiologic response to the presence of the growing conceptus, including alterations in maternal vascular endothelial cells that contribute to a profound fall in total systemic vascular resistance. There is a large body of evidence that adverse changes in the vascular endothelium underlie the multisystemic maternal manifestations of preeclampsia. Our knowledge is incomplete regarding the mechanisms of adaptive endothelial changes of normal pregnancy and why these changes are attenuated or fail in women who develop preeclampsia. Populations of bone-marrow derived endothelial progenitor cells (EPCs) exist in the adult that are mobilized into the circulation by stimuli such as estrogen and vascular endothelial growth factor. These EPCs can then differentiate into endothelial cells lining the lumen of blood vessels and/or release growth factors that act in a paracrine fashion to support the endothelium. EPCs are thus thought to function as a cellular reservoir to replace dysfunctional or senescent endothelial cells, and therefore may be critical to the overall health of the vascular endothelium. Data are emerging to suggest that the number of EPCs in the maternal circulation increases with normal pregnancy and that this change fails to occur in women with preeclampsia. While speculative at this point, our overall hypothesis is that an excess of antiangiogenic factors [such as the soluble receptors, soluble fms-like tyrosine kinase (sFlt-1) and soluble endoglin] interfere with nitric oxide-driven mobilization or activity of EPCs in the maternal circulation, contributing to the widespread endothelial dysfunction underlying the clinical manifestations of preeclampsia. PMID- 17127249 TI - Angiogenic factors and preeclampsia. AB - Preeclampsia is a major cause of maternal and neonatal morbidity and mortality worldwide. Although the etiology of preeclampsia is still unclear, recent studies suggest that its major phenotypes, hypertension and proteinuria, may be due to an excess of circulating anti-angiogenic growth factors, most notably soluble fms like tyrosine kinase 1 (sFlt1) and soluble endoglin (sEng). sFlt1 is an endogenous protein that is produced by the placenta. sFlt1 is able to bind to the angiogenic growth factors vascular endothelial growth factor and placental growth factor, thereby neutralizing their functions. High serum concentrations of sFlt1 and low concentrations of free vascular endothelial growth factor and free placental growth factor have been observed during and prior to clinical manifestation of preeclampsia. More recently, serum levels of sEng were also shown to be significantly elevated in preeclamptic women and levels of sEng correlated strongly with disease severity. Therefore, measurement of sFlt1 and sEng in the maternal circulation may be a useful diagnostic and screening tool for preeclampsia. The availability of such a test to predict preeclampsia would have significant impact on current obstetrical care and may help reduce preeclampsia-induced morbidity and mortality. This review will focus on the role of angiogenic factors in normal and abnormal placental development and indicate how measurement of circulating angiogenic factors may help identify women at risk of preeclampsia. PMID- 17127250 TI - Emerging concepts in preeclampsia investigation. AB - Preeclampsia is a disorder that uniquely affects pregnancy and profoundly alters the short- and long-term health of the mother and fetus. The pathophysiologic processes that underlie preeclampsia can be thought of in two stages: stage 1, reduced placental perfusion, and stage 2, the maternal clinical syndrome. Multiple pathophysiologic processes influence stage one, affecting trophoblast invasion and placental function. These processes in some women result in stage two, with subsequent maternal inflammatory, metabolic, and thrombotic responses, converging to alter vascular and endothelial health. An increasingly comprehensive understanding of these factors is emerging, which we will introduce in this chapter and which will be detailed in the chapters that follow. The translation of this understanding into clinical trials and ultimately into effective preventive and therapeutic measures remains the ultimate goal of preeclampsia research. PMID- 17127251 TI - Endothelial barrier function in preeclampsia. AB - Impaired endothelial barrier function resulting in increased vascular permeability is a characteristic vascular response in preeclampsia, a hypertensive and multiple systemic disorder of human pregnancy. During the last two decades, endothelial function in preeclampsia has been intensively studied and significant progress has been made in understanding the cellular and molecular bases of the altered endothelial cell response. In this review, we address the nature and mechanisms that are proposed to underlie the disturbed endothelial barrier function in preeclampsia and discuss the potential relevance of the endothelial cell responses to the initiation and/or progression of this vascular syndrome. Insights gained from the characterization of the endothelial cell phenotype assumed by the preeclamptic microvasculature may lead to novel therapeutic strategies for the management of this syndrome. PMID- 17127252 TI - Renal function during normal pregnancy and preeclampsia. AB - Glomerular filtration rate and renal plasma flow increase by 40 to 65 and 50 to 85%, respectively, during normal pregnancy in women. Studies using the gravid rat as a model have greatly enhanced our understanding of mechanisms underlying these remarkable changes in the renal circulation during gestation. Hyperfiltration is largely due to increased renal plasma flow, the latter attributable to profound reductions in both the renal afferent and efferent arteriolar resistances. The ovarian hormone, relaxin, mediates renal vasodilation during pregnancy. Relaxin increases vascular gelatinase activity, thereby converting big ET to ET(1-32), which leads to renal vasodilation, hyperfiltration and reduced myogenic reactivity of small renal arteries via the endothelial ET(B) receptor and nitric oxide. Serum concentration of uric acid falls during normal pregnancy as a consequence of increased GFR and/or reduced proximal tubular reabsorption. The elevated urinary excretion of protein during pregnancy is secondary to increased GFR, reduced proximal tubular reabsorption, and perhaps alteration in the electrostatic charge of the glomerular filter. Whether the tubular secretion of Tamm-Horsfall protein increases during normal pregnancy is uncertain. In most women with preeclampsia, renal plasma flow and glomerular filtration rate are at most only modestly decreased as a consequence of increased afferent arteriolar resistance and/or reduced ultrafiltration coefficient. Serum uric acid concentrations are increased mainly as a consequence of reduced renal clearance. Reduced GFR leads to decreased filtered load of uric acid, and plasma volume contraction contributes to increased proximal tubular reabsorption coupled to sodium. The increase in urinary protein excretion in preeclampsia occurs secondary to alterations in the size and/or charge selectivity of the glomerular filter, possible increases in glomerular capillary pressure, and compromise of proximal tubular reabsorption. The renal histologic lesion characteristic of preeclampsia is termed "glomerular endotheliosis". Recent evidence suggests that anti-angiogenic factors emanating from the placenta in preeclampsia contribute to glomerular endotheliosis, proteinuria, and hypertension during disease. PMID- 17127253 TI - Sodium regulation, sodium pump function and sodium pump inhibitors in uncomplicated pregnancy and preeclampsia. AB - Preeclampsia is a disease characterized by hypertension and proteinuria but can manifest many abnormalities. Some of the best documented alterations involve changes in the handling of sodium ion both on the systemic and on the cellular level. There is broad agreement that the components of the renin-angiotensin aldosterone pathway are markedly reduced in women with preeclampsia. However, other changes, especially those involving cell sodium are less consistent. A majority of studies support an increase in peripheral cell sodium concentration. This would suggest a defect in (Na,K)ATPase or sodium pump activity. Direct study of cellular sodium pump activity provides suggestive but not unequivocal support for this decreased sodium pump activity. Other evidence indicates increased circulating concentrations of a sodium pump inhibitor in most, but not all, studies of preeclampsia. Together, current research argues more strongly in favor of derangements of cell sodium handling perhaps mediated by circulating sodium pump inhibitors leading often to increased cell sodium. Such an increase of cell sodium in vascular tissue has previously been shown to enhance vascular sensitivity to vasoconstrictor agents or lead directly to increased vasoconstriction. PMID- 17127254 TI - Trophoblast biology, responses to hypoxia and placental dysfunction in preeclampsia. AB - Preeclampsia is a hypertensive disorder unique to human pregnancy that can result in significant morbidity and mortality for mother and fetus. While the etiology of preeclampsia is unknown, the placenta in general and trophoblast in particular is a prerequisite for the disease. We overview normal development of the human placenta, describe the role of hypoxia and other insults in placental injury, and highlight how the dysregulation of villous trophoblast biology in the second half of pregnancy may incite the pathophysiology of preeclampsia in the mother. PMID- 17127255 TI - Maternal-fetal metabolism in normal pregnancy and preeclampsia. AB - Adaptation to pregnancy in humans involves major anatomic, physiologic and metabolic changes in the mother in order to support and provide for the nutritional and metabolic needs of the growing conceptus. Metabolically, pregnancy is marked by several important and dynamic adjustments including increased insulin resistance, hyperlipidemia and changes in protein and amino acid metabolism. In general, these metabolic adaptations serve to increase nutrient availability for the benefit of the growing fetal-placental unit. Interestingly, the pregnancy complication preeclampsia is recognized to evidence biologic exaggerations of these normal metabolic adaptations of pregnancy. Specifically, preeclampsia is associated with increased insulin resistance, hypertriglyceridemia, high circulating free fatty acids, low high-density lipoprotein particles, and high maternal and fetal plasma amino acid concentrations. These metabolic alterations may contribute to the pathophysiology of the syndrome and may also influence fetal growth. The focus of this review will be to summarize the normal metabolic adaptations, transport and utilization of carbohydrates, lipids and amino acids that occur during pregnancy. Furthermore, we will review the differences in carbohydrate, lipid and amino acid metabolism in pregnancies complicated by preeclampsia in comparison to uncomplicated pregnancies. PMID- 17127256 TI - Smoking and risk of preeclampsia: a systematic review. AB - Cigarette smoking adversely affects every organ system. Paradoxically, smoking during pregnancy has been associated with a reduced risk of preeclampsia. We reviewed previous epidemiologic and clinical studies on the association between smoking and preeclampsia from 1959 to March, 2006. A total of 48 epidemiologic studies were identified. Overall, smoking during pregnancy reduces the risk of preeclampsia by up to 50% with a dose-response pattern. A protective effect was consistently found in both nulliparas and multiparas, singleton and multifetal pregnancies, and for mild and severe preeclampsia. Evidence on whether quitting smoking before or in early pregnancy reduces the risk remains inconclusive. To understand possible biologic mechanism(s) of the protective effect, we reviewed literature on potential pathophysiology of smoking and its effects on placenta, cardiovascular and immune systems. Although current literature does not lend clear evidence to support a particular mechanism for the protective effect of smoking, smoking might have effects on angiogenic factors, endothelial function and the immune system which act to lower risk of preeclampsia. More epidemiologic studies with biochemically confirmed smoking status and laboratory studies with a focus on promising pathways are warranted to further clarify this puzzling relationship. Understanding the underlying mechanisms through which smoking reduces preeclampsia risk may enhance our understanding of the pathogenesis of this disorder and contribute to the development of prevention strategies. PMID- 17127257 TI - Matrix metalloproteinases: control of vascular function and their potential role in preeclampsia. AB - Preeclampsia is a pregnancy specific disorder characterised by hypertension and proteinuria occurring after the twentieth week of gestation. Preeclampsia induced hypertension is the result of increased vascular reactivity and endothelial dysfunction, however, the mechanisms underlying this state remain elusive. One possible mediator may be the matrix metalloproteinases (MMPs), a family of proteinases typically recognized for long term tissue remodelling. This review examines the evidence suggesting a role for MMPs in acutely regulating vascular function. Studies have shown that MMPs can activate vasoconstrictors (e.g. endothelin), inactivate vasodilators (e.g. calcitonin gene related peptide) and transactivate cell surface receptors responsible for vasoconstriction (e.g. epidermal growth factor receptor). The potential role of these proteinases in preeclampsia will then be discussed. PMID- 17127258 TI - Plant lipidomics: discerning biological function by profiling plant complex lipids using mass spectrometry. AB - Since 2002, plant biologists have begun to apply mass spectrometry to the comprehensive analysis of complex lipids. Such lipidomic analyses have been used to uncover roles for lipids in plant response to stresses and to identify in vivo functions of genes involved in lipid metabolism. PMID- 17127259 TI - Gene therapy in cutaneous wound healing. AB - Medical treatment of complicated acute and chronic wounds represents a persistent and increasing medical and economic problem in our health-care system. In this review, we discuss the potentials and limitations of current and future gene therapy for the treatment of complicated, acute and chronic wounds. Chronic non healing wounds result in significant morbidity, prolonged hospitalization, lost time from work and enormous health-care expenses. There are constant efforts to improve the therapeutic modalities to local treatment of wounds. One of them is gene-therapy where the delivery of peptides directly into the wound provides a relatively new and exciting possibility. The two groups of peptides of particular interest are growth-factors and anti-microbial peptides. Gene delivery of these peptides provides not only the possibility of more targeted local delivery but also larger concentrations. Many new techniques for gene delivery to wounds have been developed in recent years. The combination of mechanical and viral or chemical vectors appear to have the greatest yield. This review provides an update on gene delivery to cutaneous wounds. PMID- 17127260 TI - Osteoclast differentiation and gene regulation. AB - Osteoclasts, the bone resorbing cells, play a key role both in normal bone remodeling and in the skeletal osteopenia of arthritis, osteoporosis, periodontal disease and certain malignancies. Osteoclast cellular commitment, differentiation and function depend upon the establishment of specific patterns of gene expression achieved through networks of transcription factors activated by osteoclastogenic cytokines. This review is an updated look at the various transcription factors and cytokines that have been demonstrated to play critical roles in osteoclast differentiation and function, along with their known animal models, such as: PU.1, Mcsf, RANKL, NF-kappaB, AP-1, NFATc1, Mitf, Myc, and Src. Further studies on these transcription factors and cytokines will not only expand our basic understanding of the molecular mechanisms of osteoclast differentiation, but will also aid our ability to develop therapeutic means of intervention in osteoclast-related diseases. PMID- 17127262 TI - B cell depletion therapy in autoimmune diseases. AB - Our understanding of the multiple physiological and pathological functions of B cells continues to expand at a fascinating rate. A critical part of this expanding knowledge is the realization that B-cells can be responsible, at least in part, for diseases in which they had not been previously suspected and that their pathogenic influence can be mediated by multiple mechanisms. In turn, the availability of effective agents capable of inducing profound and long-lasting B cell depletion and the safety and efficacy of Rituximab in non-Hodgkin lymphoma has prompted investigators to use this therapeutic approach in a large number of autoimmune diseases. Thus far, the results have been very promising, and in some cases nothing short of spectacular. In this review, we shall discuss the roles of B-cells in health and disease and the available evidence regarding the efficacy of B-cell depletion in human autoimmunity. Finally, we will discuss some of the many challenges and opportunities that the medical and scientific community should address in the foreseeable future. PMID- 17127263 TI - MALDI-TOF MS in lipidomics. AB - So far, matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) seemed to be nearly a synonym for protein analysis. However, there is growing evidence that this technique is also an useful tool in lipid analysis and lipidomics because of its fast, simple and convenient performance allowing to record mass spectra of cells, crude tissue or body fluid extracts or even intact tissue slices in a few minutes. On the negative side, however, the reproducibility of MALDI-TOF mass spectra depends significantly on the homogeneity of the co-crystals between matrix and analyte and different lipid classes are detected with different sensitivities. This is especially important because lipids with quaternary ammonia groups (e.g., GPCho) may prevent the detection of other lipid classes (e.g., GPEtn). This review starts with a short overview on traditional methods of lipid analysis with the focus on mass spectrometric methods and compares MALDI-TOF MS with other important ionization techniques. Afterwards, some landmarks in the development of MALDI-TOF MS will be introduced and some important examples in the field of tissue and body fluid lipid analysis will be discussed. This review ends with a short outlook and summary focusing on the advantages and drawbacks of MALDI-TOF MS in lipidomics. PMID- 17127261 TI - Roles of transglutaminases in cardiac and vascular diseases. AB - All transglutaminases share the common enzymatic activity of transamidation, or the cross-linking of glutamine and lysine residues to form N epsilon (gamma glutamyl) lysyl isopeptide bonds. The plasma proenzyme factor XIII is responsible for stabilizing the fibrin clot against physical and fibrinolytic disruption. Another member of the transglutaminase family, tissue transglutaminase or TG2 is abundantly expressed in cardiomyocytes, vascular cells and macrophages. The transglutaminases have a variety of functions independent of their transamidating activity. For example, TG2 binds and hydrolyzes GTP, thereby fostering signal transduction by several G protein coupled receptors. Accumulating evidence points to novel roles for factor XIII and TG2 in cardiovascular biology including: (a) modulating platelet activity, (b) regulating glucose control, (c) contributing to the development of hypertension, (d) influencing the progression of atherosclerosis, (e) regulating vascular permeability and angiogenesis (f) and contributing to myocardial signaling, contractile activity and ischemia/reperfusion injury. In this review, we summarize the cardiovascular biology of two members of the family of transglutaminases, Factor XIII and TG2. PMID- 17127264 TI - Phytate acts as an inhibitor in formation of renal calculi. AB - The aim of this study was to assess the inhibitory action of phytate in formation of renal calculi. Hypertension (induced by nicotine) combined with hypercalcemia (induced by D vitamin) was used to induce calcification in renal tissue in male Wistar rats that were fed a purified phytate free diet. Phytate non-treated rats developed significant calcium deposits in kidneys and papillae, as well as in kidney tubules and vessels, whereas calcium deposits were absent in control and phytate treated rats. Fragments of hydroxyapatite (HAP) calculi exhibited the capacity to induce the growth of calcium salts on their surfaces. Presence of 1.5 mg/L of phytate in the synthetic urine inhibited the formation of calcium oxalate monohydrate on HAP renal calculi in normocalciuric conditions. The findings show that the action of phytate as a crystallization inhibitor takes place both in the intrapapillary tissue and urine. PMID- 17127265 TI - Molecular chaperones: multiple functions, pathologies, and potential applications. AB - Cell stressors are ubiquitous and frequent, challenging cells often, which leads to the stress response with activation of anti-stress mechanisms. These mechanisms involve a variety of molecules, including molecular chaperones also known as heat-shock proteins (Hsp). The chaperones treated in this article are proteins that assist other proteins to fold, refold, travel to their place of residence (cytosol, organelle, membrane, extracellular space), and translocate across membranes. Molecular chaperones participate in a variety of physiological processes and are widespread in organisms, tissues, and cells. It follows that chaperone failure will have an impact, possibly serious, on one or more cellular function, which may lead to disease. Chaperones must recognize and interact with proteins in need of assistance or client polypeptides (e.g., nascent at the ribosome, or partially denatured by stressors), and have to interact with other chaperones because the chaperoning mechanism involves teams of chaperone molecules, i.e., multimolecular assemblies or chaperone machines. Consequently, chaperone molecules have structural domains with distinctive functions: bind the client polypeptide, interact with other chaperone molecules to build a machine, and interact with other complexes that integrate the chaperoning network. Also, various chaperones have ATP-binding and ATPase sites because the chaperoning process requires as, a rule, energy from ATP hydrolysis. Alterations in any one of these domains due to a mutation or an aberrant post-translational modification can disrupt the chaperoning process and cause diseases termed chaperonopathies. This article presents the pathologic concept of chaperonopathy with examples, and discusses the potential of using chaperones (genes or proteins) in treatment (chaperonotherapy). In addition, emerging topics within the field of study of chaperones (chaperonology) are highlighted, e.g., genomics (chaperonomics), systems biology, extracellular chaperones, and anti-chaperone antibodies. PMID- 17127266 TI - Neurolipidomics: challenges and developments. AB - The field of lipidomics is one of the most rapidly expanding areas of systems biology research. Considering the uniqueness and complexity of the lipidome in the nervous system (i.e., neurolipidome), neurolipidomics remains quite challenging but exciting. With the recent development of mass spectrometry (MS) based lipidomics, particularly the rapid improvement of multi-dimensional MS based shotgun lipidomics, much progress has been made in neurolipidomics. As the accelerated development of future technologies enables lipidomics penetrance into lower and lower abundance regions of mass contents of individual lipid molecular species, it can be anticipated that many biochemical mechanisms underlying lipid metabolism critical to neuronal disease states will be increasingly uncovered. Through exploiting the information content inherent in the complexity of neuronal lipid composition and kinetic turnover which can be revealed by neurolipidomics, substantial insights into neuronal plasticity and gene function can be gathered. Through neurolipidomics, the markers for these neuronal diseases which identify pathological alterations and are diagnostic of disease onset, progression or severity may potentially be discovered. Accordingly, with neurolipidomics, our understanding of the complexities of the nervous system will be undoubtedly accelerated, as many mysteries are resolved. PMID- 17127267 TI - Ethanol intake enhances inflammatory mediators in brain: role of glial cells and TLR4/IL-1RI receptors. AB - The brain is one of the major target organs of ethanol actions, and its chronic and acute intoxication results in significant alterations in brain structure and function, and in some cases to neurodegeneration. Glial cells and Toll-like receptors (TLRs) are vital players in CNS immune response; dysregulation of this response plays an important role in brain damage and neurodegeneration. Ethanol has immunomodulatory effects and induces specific alterations in the TLRs response in many tissues. These actions depend on the cell type, ethanol dose and treatment duration, as well as the concomitant presence of pathogens and their characteristics. Recent findings indicate that low concentrations of ethanol (10 mM) promote inflammatory processes in brain and in glial cells by up-regulating cytokines and inflammatory mediators (iNOS, NO, COX-2), and by activating signaling pathways (IKK, MAPKs) and transcriptional factors (NF-kappaB, AP-1) implicated in inflammatory injury. TLR4/IL-1RI receptors may be involved in ethanol-mediated inflammatory signaling, since blocking these receptors abolishes the production of ethanol-induced inflammatory mediators and cell death. We propose that at low physiologically relevant concentrations, ethanol facilitates TLR4/IL-1RI recruitment into lipid rafts microdomains, leading to the activation and signaling of these receptors. In summary, current results suggest that TLR4/ IL-1RI are important targets of ethanol-induced inflammatory brain damage. PMID- 17127268 TI - What triggers cell-mediated mineralization? AB - Mineralization is an essential requirement for normal skeletal development, but under certain pathological conditions organs like articular cartilage and cardiovascular tissue are prone to unwanted mineralization. Recent findings suggest that the mechanisms regulating skeletal mineralization may be similar to those regulating pathological mineralization. In general, three forms of cell mediated mineralization are recognized in an organism: intramembranous ossification, endochondral ossification and pathological mineralization. This review summarizes recent work that tried to elucidate how cell-mediated mineralization is initiated and regulated. To explain mineralization, several theories have been proposed. One theory proposes that mineralization is initiated within matrix vesicles (MVs). A second, not mutually exclusive, theory proposes that phosphate induces apoptosis, and that apoptotic bodies nucleate crystals composed of calcium and phosphate. A third theory suggests that mineralization is mediated by certain non-collagenous proteins, which associate with the extracellular matrix. Regardless of the way mineralization is initiated, the organism also actively inhibits mineralization by specific proteins and removal of an inhibitor may also induce mineralization. Although many studies greatly contributed to a better understanding of the mechanisms regulating cell-mediated mineralization, many questions remain about the mechanisms that trigger cell mediated mineralization and how this process is regulated. Further investigation is necessary to develop in the future novel therapeutic strategies to prevent pathological mineralization. PMID- 17127269 TI - Two-versus one photon excitation laser scanning microscopy: critical importance of excitation wavelength. AB - It is often anticipated that two-photon excitation (TPE) laser scanning microscopy should improve cell survival and tissue penetration relative to conventional one-photon excitation (OPE) confocal scanning laser microscopy (CLSM). However few studies have directly compared live cell imaging using one- vs two-photon laser scanning microscopy. We have used calcein-loaded in situ chondrocytes within cartilage as a model for quantitatively comparing these techniques. TPE reduced photo-bleaching and improved cell viability compared to OPE. Using improved detection sensitivity coupled with increased tissue penetration of the near infra-red TPE laser, it was possible to capture images deeper within the cartilage. However, the advantages of TPE vs OPE were strongly dependent on excitation wavelength. We conclude that optimising TPE conditions is essential for realizing the full benefits of this approach. PMID- 17127270 TI - Bacterial DNA microarrays for clinical microbiology: the early logarithmic phase. AB - In this era of coexistence of high-throughput sequencing technologies and serious difficulties in the management of both common and novel infectious syndromes, new techniques which improve the study of micro-organisms is timely. In bacteriology, the most important subjects are bacterial pathogenicity, discovery of the genomic complexity of bacteria, and the epidemiology of antimicrobial resistance traits. From the clinical point of view, genetic testing is flanking phenotypic testing for the assessment of new, difficult to test antibiotic resistance traits, and for correlations with the microbial behaviour in vivo. The demand for faster, comprehensive and highly parallel microbial diagnostics is also cogent even at the basic laboratory level, where the ultimate objective is saving lives. In this setting, DNA microarrays offer a pivotal contribution by allowing performance of hybridization experiments in highly parallel formats, with an increasing reliability. Not only they are useful in deciphering host and microbial pathophysiology, they can also make the difference in the management of prognostic and therapeutic aspects of many diseases. Here, we provide an overview of the current use and the potential of DNA microarrays in clinical bacteriology, and several applications and technical solutions are discussed. PMID- 17127271 TI - Genetic basis of respiratory distress syndrome. AB - Respiratory distress syndrome (RDS) is a multifactorial developmental disease caused by lung immaturity and presenting as high-permeability lung edema ("hyaline membrane disease"). It is characterized by a transient deficiency of alveolar surfactant during the first week of life. During the first few days of life, the alveolar surfactant pool size increases up to that in the controls. The allelic variants of the genes encoding the surfactant proteins (SP) SP-A1, SP-A2, SP-B, and SP-C have been associated with RDS. The main SP-A haplotype, interactively with the SP-B Ile131Thr polymorphism and with constitutional and environmental factors, influence the risk. Case reports on mutations with partially functional SP-B have been published. The genetic susceptibility factors depend on the degree of prematurity at birth, consistent with sequential differentiation of the lung and gestation-dependent differences in clinical presentation. The preferentially type 2 cell expressed genes involved in critical functions (such as ATP-binding cassette transporter, ABCA3), those involved in susceptibility to acute lung damage, and those with known susceptibility to other severe lung diseases (such as G protein-coupled receptor for asthma susceptibility, GPR154 alias GPRA) will possibly serve as candidate genes in future studies. RDS associated with near-term and term births may have a different genetic predisposition and pathogenesis compared to RDS after very preterm birth. As we learn more about the molecular consequences of allelic variation, new therapies based on a new generation of surfactant diagnostics and individualized therapies may follow. PMID- 17127273 TI - Aberrant spindle assembly checkpoint in bovine somatic cell nuclear transfer oocytes. AB - Nuclear, microtubular dynamics and spindle assembly checkpoint (SAC) in bovine somatic cell nuclear transfer (SCNT) oocytes receiving G1/0 or M phase somatic cell nuclei were studied. SCNT oocytes assembled microtubules, however, the spindles were structurally abnormal, including bi-, tri-polar or elongated spindles with scattered premature chromosome condensation (PCC) in G1/0 phase nuclei, and some miniature spindles with unaligned chromosomes in M phase nuclei. In contrast, demecolcine-treated SCNT oocytes formed chromosome clusters with membrane protrusion and significantly induced maturation-promoting factor (MPF) activity elevation (up to 177%) for 3 hours, indicating that first SAC at second metaphase (MII) is established upon spindle disruption in SCNT oocytes. After parthenogenetic stimuli, unlike MII oocytes which prevent exit from MII arrest with high MPF activity upon spindle disruption by second SAC, demecolcine-treated SCNT oocytes could not prevent exit from MII arrest with inactivation of MPF activities, whereas MG132-treated SCNT oocytes could persist at MII arrest, indicating that SCNT oocytes lack the ability for second SAC establishment, however, two G1/0 phase nuclei in an ooplasm restored second SAC establishment upon spindle disruption. Furthermore, the developmental potential of demecolcine treated SCNT oocytes receiving G1/0 phase nuclei to blastocyst stage was not significantly different than untreated SCNT oocytes (29% vs 31%). These results indicate that unlike MII oocytes, SCNT oocytes have aberrant spindle morphology and SAC at MII due to insufficient SAC signals from somatic cell nuclei, thus aberrant remodeling has started immediately after somatic cell nuclear transfer and may be responsible for chromosome instability in SCNT embryos as well as the low successful efficiency of cloning. PMID- 17127272 TI - The role of the activated macrophage in clearing Listeria monocytogenes infection. AB - Macrophage activation often contributes to the strong immune response elicited upon infection. The ability of macrophages to become activated was discovered when sub-lethal primary infections of mice with the bacterium Listeria monocytogenes provided protection against secondary infections through non humoral immunity. L. monocytogenes infect and propagate in macrophages by escaping the phagosome into the cytosol, where they avoid humoral immune mediators. Activated macrophages kill L. monocytogenes by blocking phagosomal escape. The timing of the antimicrobial activities within the phagosome is crucial to the outcome. In non-activated macrophages, bacterial factors generally prevail, and L. monocytogenes can escape from the vacuoles and grow within cytoplasm. Activated macrophages generate reactive oxygen or nitrogen intermediates early after bacterial uptake, which prevent the bacteria from escaping vacuoles into cytoplasm. The heterogeneity in the interactions between L. monocytogenes and the macrophage indicate a complex relationship between the host and the pathogen governed by chemistries that promote and inhibit escape from vacuoles. This review examines the mechanisms used by activated and non activated macrophages to kill microbes, and how those mechanisms are employed against L. monocytogenes. PMID- 17127274 TI - Placental cytokines and preeclampsia. AB - Preeclampsia is a serious and life-threatening pregnancy complication. Reduced uteroplacental perfusion and oxygen tension, impaired trophoblast differentiation and invasion, and altered placental production of immunomodulators and growth factors are all considered to be important aspects in the aetiology of the condition. The placenta expresses a variety of pro and anti-inflammatory cytokines, adipokines and cytokine-like angiogenic growth factors, production of which is altered in preeclampsia, driven (at least in part) by hypoxia. Altered levels of cytokines have been measured in the circulation of women with preeclampsia, although for reasons that are not always apparent much of the data are disturbingly inconsistent. While the placenta undoubtedly makes an important contribution to plasma cytokine levels, production by maternal peripheral blood mononuclear cells (PBMCs) and other tissues is also likely to be significant, although to what extent remains undetermined. Increased placental expression of soluble receptors occurs with preeclampsia, resulting in elevated circulating concentrations which confer impaired angiogenesis, deficient placental vascularisation, increased placental apoptosis and endothelial dysfunction. The extent to which these changes reflect a response to the disorder, as opposed to being a causative factor in the development of maternal disease, is a matter of some debate. Nevertheless, convincing evidence is now accruing that autocrine/paracrine interactions between placental cytokines/growth factors and the maternal endothelium play a central role in the pathogenesis of preeclampsia. PMID- 17127275 TI - Intranasal administration with NAD+ profoundly decreases brain injury in a rat model of transient focal ischemia. AB - Excessive poly(ADP-ribose) polymerase-1 (PARP-1) activation plays a significant role in ischemic brain damage. Increasing evidence has supported the hypothesis that PARP-1 induces cell death by depleting intracellular NAD+. Based on our in vitro finding that NAD+ treatment can abolish PARP-1-mediated cell death, we hypothesized that NAD+ administration may decrease ischemic brain injury. In this study, we used a rat model of transient focal ischemia to test this hypothesis. We observed that intranasal NAD+ delivery significantly increased NAD+ contents in the brains. Intranasal delivery with 10 mg/kg NAD+ at 2 hours after ischemic onset profoundly decreased infarct formation when assessed either at 24 or 72 hours after ischemia. The NAD+ administration also significantly attenuated ischemia-induced neurological deficits. In contrast, intranasal administration with 10 mg/kg nicotinamide did not decrease ischemic brain damage. These results provide the first in vivo evidence that NAD+ metabolism is a new target for treating brain ischemia, and that NAD+ administration may be a novel strategy for decreasing brain damage in cerebral ischemia and possibly other PARP-1-associated neurological diseases. PMID- 17127276 TI - Potential role of humoral immunity in the pathogenesis of multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE). AB - The earliest research literature addressing sub-clinical characteristics of Multiple Sclerosis was largely focused on humoral immune components, particularly antibodies in the cerebrospinal fluid of MS patients. However, two decades later, in the 1990's, T cells were established as a major component of the underlying mechanism(s) of MS pathogenesis, especially since EAE, the mouse model of MS, could be readily induced by immunization with myelin derived peptides or passive transfer of encephalogenic T cells. This data has contributed to the concept that the role of humoral immunity in MS pathogenesis may be negligible. However, more recent studies have provided important insights regarding the role of humoral immunity in MS pathogenesis. The goals of this review are to 1) summarize evidence for and against the hypothesis that humoral immunity plays a central role in the pathogenesis of MS, and 2) summarize studies in the EAE model that directly tested the role of humoral immunity in pathogenesis of the disease. With this information, we hope to convince the reader that great strides have been made towards defining a central role of humoral immunity in MS pathogenesis, but that there is a substantial amount of work to be done (especially in the EAE model) to ensure that the contribution of humoral immunity to MS pathogenesis is effectively addressed. PMID- 17127277 TI - Myocardial lipidomics. Developments in myocardial nuclear lipidomics. AB - The development of electrospray ionization mass spectrometry has been critical for the analyses of lipidomes from subcellular organelles. The myocardial nuclear lipidome likely has a key role in the molecular regulation of gene expression. In fact, recent studies have suggested that specific phospholipid classes bind and regulate specific transcription factors. The dynamic regulation of the myocardial nuclear lipidome may be critical in mediating long-term pathological responses to stresses such as ischemia, tachycardia, and hypertension. In this brief review, the preparation of myocardial nuclei is discussed, and the resulting nuclear lipidome from rat and rabbit are shown as examples. The rabbit myocardial nuclear lipidome contains relatively more plasmenylcholine/phosphatidylcholine molecular species in comparison to that ratio observed in the rat myocardial nuclear lipidome. The composition of the rat myocardial nuclear choline glycerophospholipid pool was relatively enriched with molecular species containing arachidonic acid and docosahexaenoic acid in comparison to that in the rabbit myocardial nuclear choline glycerophospholipid pool. While the ethanolamine glycerophospholipids of the rabbit myocardial nuclei are enriched with arachidonic acid and plasmalogens, the ethanolamine glycerophospholipid profile from rat myocardial nuclei show less plasmalogen and more species containing docosahexaenoic acid. Last, significant differences in the ethanolamine glycerophospholipid molecular species were observed in the rabbit heart lipidomes from the nucleus and the mitochondria. Quantitation of these lipid species in hearts subjected to pathophysiological stresses may provide important information on the role of the myocardial nuclear lipidome on long-term cardiac cell function. PMID- 17127278 TI - Role of TWEAK and Fn14 in tumor biology. AB - The tumor necrosis factor (TNF) superfamily member TNF-like weak inducer of apoptosis (TWEAK) was initially described in a 1997 publication co-authored by investigators from the biotechnology company Biogen (now Biogen-Idec) and the University of Geneva. Four years later, researchers at the biotechnology company Immunex (now part of Amgen) reported the cloning and characterization of the human TWEAK receptor. A sequence database search revealed that the predicted TWEAK receptor amino acid sequence was identical to that of fibroblast growth factor-inducible 14 (Fn14), a small transmembrane protein described one year earlier in a publication from investigators at the American Red Cross Holland Laboratory. Recent studies have revealed that the TWEAK-Fn14 ligand-receptor pair likely plays an important role in a variety of cellular processes and in the pathogenesis of several human diseases, including atherosclerosis, stroke, rheumatoid arthritis and cancer. In this paper, we first summarize the general properties of these two proteins and then review the available data implicating TWEAK and Fn14 in multiple aspects of tumor biology. PMID- 17127279 TI - Cell fusion: biological perspectives and potential for regenerative medicine. AB - Cell fusion has emerged as a powerful subject of debate in the last few years. Adult stem cell plasticity and the search for mechanisms to explain this process have led to the "rediscovery" of cell fusion. In nature, cell fusion is a normal process involved in sexual reproduction, tissue formation, and immune response. The recent observation that bone marrow derived cells fuse with several cell types introduces new and provocative questions. In this review, I shall recapitulate what is known about cell fusion and discuss its more controversial aspects. I shall highlight the most exciting open questions; its biological potential; pros and cons; and their implications on stem cell plasticity, regenerative medicine, and development. PMID- 17127280 TI - Animal models for the study of liver regeneration: role of nitric oxide and prostaglandins. AB - The mechanisms that permit adult tissues to regenerate are the object of intense study. Liver regeneration is a research area of considerable interest both from pathological and from physiological perspectives. One of the best models of the regenerative process is the two-thirds partial hepatectomy (PH). After PH, the remnant liver starts a series of timed responses that first favor cell growth and then halts hepatocyte proliferation once liver function is fully restored. The mechanisms regulating this process are complex and involve many cellular events. Initiation of liver regeneration requires the injury-related cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin 6 (IL-6), and involves the activation of cytokine-regulated transcription factors such as NF-kappaB and STAT3. An important event that takes place in the hours immediately after PH is the induction of nitric oxide synthase 2 (NOS-2) and cyclooxygenase 2 (COX-2), and the consequent release of nitric oxide (NO) and prostaglandins (PGs). NO is involved in the vascular readaptation after PH, favoring a general permeability to growth factors throughout the organ. This review examines the mechanisms that regulate NO release during liver regeneration and the animal models used to identify these pathways. PMID- 17127281 TI - Genetically engineered mice as a model for studying cardiac arrhythmias. AB - Sudden cardiac death due to ventricular tachyarrhythmias remains an unresolved problem, probably because the mechanisms responsible for the progression of cardiac disease to electrophysiological failure are poorly understood. Genetically engineered mice, the principal mammalian model for studying cardiac electrophysiology, have contributed to the understanding of the genetic, molecular and systemic mechanisms involved in the initiation and/or maintenance of cardiac arrhythmias leading to cardiac death, e.g. cardiac excitability, conduction velocity and refractoriness. Several murine models harbouring human gene mutations leading to electrical and structural cardiac disorders have been developed, including channelopathies (long QT syndrome), familial conduction disorders, cardiomyopathies and other inherited cardiac disorders. This article reviews the results of the main genetically modified mice addressing the genesis of cardiac arrhythmias and sudden cardiac death. PMID- 17127282 TI - Age-related cardiac deterioration: insights from Drosophila. AB - As the average lifespan in Western countries continues to expand, health care for the aged has become an increasingly important research focus. While clinicians and vertebrate researchers have frequently concentrated on specific age-related diseases, particularly neurodegenerative diseases such as Alzheimer's and Parkinson's Diseases, researchers working with invertebrate genetic model systems have gained important insights into global mechanisms of lifespan determination. Still others have employed biochemical and molecular approaches to elucidate processes contributing to common diseases of the elderly, such as cancer and diabetes. In between the broad focus on organismal aging and the more narrow focus on cellular dysfunction is the study of aging at the level of individual organ function. This review will attempt to highlight recent advances in the area of age-related deterioration of organ function provided by the use of transgenic model organisms, with a view toward incorporating these observations into a framework provided by both broader theories of the aging process and studies of cellular function during aging. PMID- 17127283 TI - The risk for cardiovascular disease in women: from estrogens to selective estrogen receptor modulators. AB - Cardiovascular disease, a generic denomination including coronary heart disease (CHD), stroke, and venous thromboembolic disease (VTED), has shown sensitivity to estrogens. The relative protection of women as compared with men has nourished a debate about a possible protective role for estrogens, but the prejudicial effects detected in clinical trials has created confusion on the risk/benefit ratio induced by hormone administration. The hypothesis that agonists distinct to estrogens might improve the effects associated with estrogens is at the base of the increasing interest on the role of selective estrogen receptor modulators (SERMs). There is a lack of definitive clearcut clinical data on the effects of SERMs in CVD, although the available information suggests a neutral balance on CHD and stroke and an increase in risk similar to estrogens for VTED. Research on pathogenetic mechanisms concentrates in atherosclerosis as the main determinant of the arterial forms of the disease and in hypercoagulability as the counterpart for venous disease. The different experimental models used up to the present moment suggest that, compared with estrogens, SERMs play a less active protection against atherogenesis but do not increase vulnerability of unstable plaques. There is not a clear notion on the mechanisms promoted by SERMs to increase risk for VTED. PMID- 17127284 TI - Isolated teratozoospermia: a cause of male sterility in the era of ICSI? AB - Single structural defects involving the totality of ejaculated sperm are among rare cases of untreatable human male infertility. This form of infertility is of genetic origin and is generally transmitted as an autosomal recessive traits. Acrosome agenesis or globozoospermia results from perturbed expression of nuclear proteins or from an altered Golgi-nuclear recognition during spermiogenesis. Failed fertilization after intracytoplasmic sperm injection (ICSI) of acrosomeless sperm is consistent with an inability of sperm to activate oocytes. Acephalic spermatozoa result from a head-neck defect due to a failure of migration of the tail anlagen and related centriole to the caudal pole of spermatids. An abnormal sperm centrosome function may explain the defective embryo cleavage after ICSI with sperm carrying a fragile head-neck junction. Primary cilia dyskinesia (PCD) and dysplasia of the fibrous sheath (DFS) are isolate defects associated with absent or greatly reduced sperm motility due to an abnormal ciliary structure and function (PCD) or to a disorganized fibrous sheath (DFS). Numerous defective genes are potentially involved in human isolated teratozoospermia but such defects have not been defined at the molecular level in most cases. IVF-ICSI is the only available method for obtaining live births with sperm carrying these defects, but the outcome is poor and the genetic risk for the subsequent generation can not be determined. PMID- 17127285 TI - Sperm phospholipases and acrosomal exocytosis. AB - At the time of fertilization, the sperm cell undergoes regulated exocytosis in response to the oocyte-associated agonists progesterone and zona pellucida. An early response generated by agonist-receptor interaction in spermatozoa is the activation of mechanisms leading to Ca2+ influx, this ion being essential for the activation of phospholipases and for the fusion of the plasma membrane with the outer acrosomal membrane. Both a phosphoinositide-specific, and a phosphatidylcholine-specific phospholipase C are involved in the generation of a variety of diacylglycerol molecular species. Phospholipase D, on the other hand, does not seem to play a significant role in the generation of diacylglycerol. Hydrolysis of phospholipids by phospholipase A2 generates free fatty acids and lysophospholipids; these are important either as substrates for the generation of other metabolites (e.g., eicosanoids) or having a direct, essential action in the final stages of membrane fusion. There is still much work to be done in the future in order to characterize phospholipase isozymes and their regulation during acrosomal exocytosis in spermatozoa. PMID- 17127286 TI - Treatment of severe male infertility by micromanipulation-assisted fertilization: an update. AB - In the past 5-10 years the evolution of micromanipulation-assisted fertilization for the treatment of severe male infertility was marked by the introduction of new technical support, refinement of diagnostic methods for the evaluation of sperm developmental potential, and development of new treatment regimens for the newly discovered abnormalities. The new technical support involves the use of non contact laser technology to assist micromanipulation for fertilization, the evolution of polarized microscopy-based optical systems to non-invasively detect the position of the meiotic spindle in living human oocytes, and the development of high-magnification optical systems for a better morphological selection of spermatozoa to be used for fertilization. Diagnostic approaches were enriched by commercial availability of kits for the analysis of sperm DNA integrity, leading to the definition of sperm nuclear DNA damage as a distinct cause of male infertility, and by the development of tests, based on heterologous ICSI, for detection of sperm failure to activate oocytes. Several treatment options for these conditions have been proposed and are currently being tested in larger scale trials. Some technical improvement was also achieved in the field of in vitro maturation of germ cells from men with in vivo maturation arrest, but only a modest clinical improvement resulted from their application. As to the risk for the offspring, recent data are rather reassuring. Except for the risk of transmission of genetically based infertility, no straightforward evidence for a health risk derived from these techniques has been provided. Nevertheless, caution is necessary, particularly concerning the eventual increase in genomic imprinting abnormalities. PMID- 17127287 TI - Genetic basis of preterm birth. AB - Epidemiologic data show that women who deliver prematurely often have a personal and/or family history of preterm birth (PTB) and that racial and ethnic differences influence the incidence of PTB. This may indicate genetic predisposition to PTB. However, since races and ethnic groups tend to share environmental factors (exposure to toxins, living conditions, diet, smoking), epidemiologic data may just confirm environmental influences on PTB. Alternatively, PTB may represent a consequence of gene-environment interactions. Infection and inflammation correlate with increased risk for preterm premature rupture of amniotic membranes (PPROM) and PTB. Immunomodulatory molecules and their receptors regulate these processes and many of them are products of polymorphic genes. Single nucleotide polymorphisms (SNPs) of a gene may lead to a differential expression of its product. So far, SNPs for several genes have been implicated in PTB. If it is confirmed that polymorphism(s) in particular gene(s) correlates with PTB, it may become possible to develop targeted diagnostic and therapeutic approaches tailored towards unique genetic characteristics of a mother/fetus pair. PMID- 17127288 TI - Genetic determinants of mammalian pituitary morphogenesis. AB - The anterior pituitary contains five trophic (hormone-secreting) cell types which are defined by their hormone products. During pituitary organogenesis, these lineages emerge in a stereotypical spatio-temporal pattern from a common ectodermal primordium, Rathke's Pouch (RP), thereby providing an excellent model system to address key developmental processes such as pattern formation, cell specification and differentiation. Genetic studies performed in mice have revealed that secreted factors released from neighbouring tissues are critical for the formation of RP and appear to establish positional identity within RP through regionally-restricted induction of transcription factor gene expression. Together, these transcription factors coordinate progenitor cell proliferation, specification and differentiation via a variety of mechanisms that include the recruitment of cell type specific co-activator and co-repressor complexes. Herein we discuss the roles of key components in the pituitary developmental program with particular focus on functionally conserved genes which are associated with various forms of pituitary hormone deficiency in humans. PMID- 17127289 TI - Copper-binding compounds as proteasome inhibitors and apoptosis inducers in human cancer. AB - The trace element copper is vital to the healthy functioning of organisms. Copper is used in a multitude of cellular activities including respiration, angiogenesis, and immune responses. Recently, copper has become a focus in medical research ranging from Alzheimer's disease to cancer. Copper modulation has been suggested to be a potential modality for therapy in these diseases. Several copper-binding compounds have been found to spontaneously complex with copper and form active proteasome inhibitors and apoptosis inducers. This review examines compounds in the quinoline and dithiocarbamate families and from the National Cancer Institute (NCI) Diversity Set that bind with copper and act as anticancer agents. In each case, it is shown that these compounds can bind with copper, inhibit the proteasome activity, and induce apoptosis in cancer cells. These activities are absent when copper is not present. Compounds alone, clioquinol and pyrrolidinedithiocarbamate as examples, are shown to have no effects in normal breast cells. Current research suggests that a possible therapeutic modality for cancer may be developed using the difference of high copper load in tumors versus low copper load in normal cells. This strategy would convert tumor cellular copper into a potent, specific proteasome inhibitor and apoptosis inducer. Thus, this approach could pave the way for the development of nontoxic anticancer therapy. PMID- 17127290 TI - Cap1p plays regulation roles in redox, energy metabolism and substance transport: an investigation on Candida albicans under normal culture condition. AB - Cap1p, a transcription factor in Candida albicans, is believed to be required for tolerance to oxidative stress. However, no information is available concerning its function on basal transcriptional profile. In this study, differentially expressed genes between the CAP1-deleted strain and its parental strain under normal culture condition were identified through microarray analysis. Notably, among the 48 down-regulated genes with the deletion of CAP1, there were three clusters, functionally related to intracellular redox, energy metabolism and substance transport. IPF7817, IPF11105 and FDH11, the three putative Cap1p target genes functionally related to redox, were shown to be activated by oxidative stress in a Cap1p-dependent manner. Furthermore, rhodamine 6G efflux analyses demonstrated that Cap1p contributed to the energy-driven efflux. Taken together, these results reveal that Cap1p plays a significant role in redox status regulation, energy metabolism and substance transport under normal culture condition. PMID- 17127291 TI - Signature of mitochondria of steroidal hormones-dependent normal and cancer cells: potential molecular targets for cancer therapy. AB - The cross-talk between the cell nucleus and mitochondria appears to control hormone-induced signaling involved in the apoptosis, proliferation, and differentiation of both normal and malignant cells. Evaluation of the defects in genetics and physiology of human endocrine diseases, such as cancer, may manifest as a result of mitochondrial physiologic and metabolic compensation of genetic defects. Steroidal agents control biogenesis and maintenance of mitochondria through the crosstalk between nuclear and mitochondrial genomes. The regulation of mitochondrial transcription by steroidal hormones, presumably occurring through pathways similar to those that take place in the nucleus, opens a new way to better understand steroid hormone and vitamin action at the cellular level. In addition to the steroid hormone receptors, estrogen generated mitochondrial oxidants together with an estrogen-driven increase in epithelial cell proliferation have been shown to participate in the initiation and promotion of the neoplastic lesions in estrogen-sensitive tissues. Mitochondria generation of ROS appears to transduce signals to the nucleus for the activation of transcription factors involved in the cell cycle progression of estrogen dependent cancer cells. Therefore, an in-depth analysis of such redox regulatory mechanisms is pertinent to the development of novel drugs and gene therapy strategies for the treatment of steroid hormone-dependent diseases related to mitochondrial disorders including cancer. PMID- 17127293 TI - Genetic polymorphism in bladder cancer. AB - Individual variation in the genetic constitution of humans may affect the host responses to constant assaults from exogenous and endogenous carcinogens, which will eventually impact cancer risk, disease prognosis and clinical outcome. Bladder cancer is one of the most common cancers in the world. In this review, the published research articles studying the association between genetic polymorphisms and bladder cancer risk and disease progression are summarized. Genetic polymorphisms are categorized based on their primary cellular functions: genes in carcinogen metabolism, DNA repair, cell cycle control, inflammation, apoptosis, methylation, genes functioning as G proteins, and cell adhesion molecules. Furthermore, we discuss a number of limitations of current genetic susceptibility research and suggest future directions in molecular epidemiology study. This review presents an overview of current molecular epidemiology of bladder cancer and provides a useful resource for understanding the pathogenesis of bladder cancer. PMID- 17127294 TI - Osteopontin: a novel inflammatory mediator of cardiovascular disease. AB - Osteopontin, also called cytokine Eta-1, is a multifunctional protein containing Arg-Gly-Asp-Ser (RGDS) cell-binding sequence. It interacts with alpha(v)beta1, alpha(v)beta3 and alpha(v)beta5 integrins and CD44 receptors. OPN is suggested to play a role during inflammation via the recruitment and retention of macrophages and T-cells to inflamed sites. OPN regulates the production of inflammatory cytokines and nitric oxide in macrophages. In this review, we will discuss diverse roles of OPN related to cardiovascular diseases, including atherosclerosis, valvular stenosis, hypertrophy, myocardial infarction and heart failure. PMID- 17127292 TI - Mechanotransduction in skeletal muscle. AB - Mechanical signals are critical to the development and maintenance of skeletal muscle, but the mechanisms that convert these shape changes to biochemical signals is not known. When a deformation is imposed on a muscle, changes in cellular and molecular conformations link the mechanical forces with biochemical signals, and the close integration of mechanical signals with electrical, metabolic, and hormonal signaling may disguise the aspect of the response that is specific to the mechanical forces. The mechanically induced conformational change may directly activate downstream signaling and may trigger messenger systems to activate signaling indirectly. Major effectors of mechanotransduction include the ubiquitous mitogen activated protein kinase (MAP) and phosphatidylinositol-3' kinase (PI-3K), which have well described receptor dependent cascades, but the chain of events leading from mechanical stimulation to biochemical cascade is not clear. This review will discuss the mechanics of biological deformation, loading of cellular and molecular structures, and some of the principal signaling mechanisms associated with mechanotransduction. PMID- 17127295 TI - Molecular pathogenesis of hepatitis C virus-associated hepatocellular carcinoma. AB - Chronic infection with hepatitis C virus (HCV) is causally associated with the development of hepatocellular carcinoma (HCC). HCV is not cytolytic and replicates entirely in the cytoplasm. Viral interaction with the host leads to subversion of immune response and other defense mechanisms. The recent development of robust cell culture systems for HCV infection provides new opportunities for the study of virus-cell interaction and viral pathogenesis. HCV infection causes active inflammation and fibrosis, which ultimately progresses to cirrhosis. The onset of cirrhosis usually precedes the multistage process of tumor development, in which common themes of viral carcinogenesis can be identified. While chronic inflammation and cirrhosis are thought to play an important role in tumor initiation, the underlying mechanisms are incompletely understood. Recent studies have revealed that infection with HCV induces genome instability, leading to further genetic and epigenetic alterations which contribute to the full development of HCC tumor. The expression of viral oncoproteins such as C and NS5A is critically involved both in the induction of genome instability and in dysregulating cellular control of growth and signal transduction. A better understanding of the molecular pathogenesis of HCV will reveal novel strategies for the prevention and treatment of related diseases including HCC. PMID- 17127296 TI - Oncogenesis and transforming viruses: the hepatitis B virus and hepatocellularcarcinoma--the etiopathogenic link. AB - Hepatocellular carcinoma (HCC) is one of the world's leading fatal malignancies. Chronic infection with the hepatitis B virus (HBV) has been implicated with the development of HCC. For the past three decades, intensive research has focused on the role of HBV in hepatocarcinogenesis. Various HBV-associated models have emerged, but increasing evidence points to two major HBV-specific mechanisms that contribute to the development of HCC. The first is the integration of the viral genome into the host chromosome causing cis-effects, resulting in loss of tumor suppressor gene functions, and/or activation of tumor-promoting genes. The second mechanism involves the expression of trans-activating factors derived from the HBV genome, which have the potential to influence intracellular signal transduction pathways and alter host gene expression. A major player involved in this form of viral transactivation is the X protein (HBx). The HBx protein was found to display pleiotropic functions and has been implicated in the malignant transformation of chronically-infected liver cells. By disrupting cellular gene expression, viral products such as HBx may modulate cellular growth, repair and death, consequently resulting in the transformation of hepatocytes to an oncogenic state. PMID- 17127297 TI - Vaccines against human papillomavirus. AB - Human papillomavirus has been identified as an etiological factor for cervical cancer, anogenital cancers and a subset of head and neck cancers. These important observations suggest that HPV vaccines have potential in the prevention and treatment of cervical cancer and other HPV-associated malignancies. The HPV genome encodes two HPV late genes, L1 and L2, which form the viral capsid. Early viral proteins support viral genome replication, two of which (E6 and E7) are important for HPV associated malignant transformation. Prophylactic HPV vaccines prevent infection by inducing neutralizing antibodies against HPV capsid proteins L1 and L2. However, because HPV-infected basal keratinocytes and HPV-transformed cells generally do not express L1 or L2, therapeutic HPV vaccines aim to treat established HPV infections and HPV-associated malignancies by targeting non structural early viral antigens of HPV such as E6 and E7. Results from preclinical HPV vaccine studies have led to several HPV vaccine clinical trials. If these prophylactic and therapeutic HPV vaccines prove as successful in patients as they have in animal models, vaccination may provide for control and eventually eradication of oncogenic HPV infection and HPV-related cancers. PMID- 17127298 TI - Human herpesvirus 8-encoded proteins with potential roles in virus-associated neoplasia. AB - Human herpesvirus 8 (HHV-8) is a gamma-2 herpesvirus, related genetically to simian herpesvirus saimiri (HVS), the prototype virus of this subgroup of the gammaherpesvirus subfamily. HHV-8 DNA is present in all forms of Kaposi's sarcoma (KS) and primary effusion lymphoma (PEL), and in most forms of multicentric Castleman's disease (MCD), especially in HIV infected individuals. Of relevance to attempts to explain the molecular basis of HHV-8 associated neoplasia, are the unique genes specified by this virus, in particular angiogenic cytokines viral interleukin-6 (vIL-6) and viral CC-class chemokines (vCCL-1, vCCL-2, vCCL-3), mitogenic signaling membrane proteins variable ITAM-containing protein (VIP) and latency associated membrane protein (LAMP), pro-survival latently-expressed viral interferon regulatory factor (vIRF3), and the kaposin family of proteins that promote cell growth and cytokine production. Also of relevance are the angiogenic and cytokine-inducing viral G protein-coupled receptor (vGPCR), pro-proliferative and pro-survival latency proteins viral FLICE inhibitory protein (vFLIP) and latency-associated nuclear antigen (LANA), and G1-S phase cell-cycle promoter viral cyclin (v-cyclin), proteins specified also by other gamma-2 herpesviruses. The enormous progress on the characterization of the properties and biological activities of these proteins over the last ten years has provided insight into the potential mechanisms of HHV-8-induced neoplasia. Present data suggest that there operates a combination of cell transformation mediated by latently expressed proteins that promote cell proliferation and survival coupled with paracrine signaling functions mediated by either the viral cytokines or viral receptor-induced secreted cellular proteins. This review discusses the properties of the viral proteins believed to contribute to viral neoplasia via these mechanisms. PMID- 17127299 TI - Gonadotropin-controlled mammal oocyte meiotic resumption. AB - Fully grown mammalian oocytes resume meiosis as a consequence of rises in gonadotropin levels at the mid-cycle. The increase of cyclic adenosine 3',5' monophosphate (cAMP) and the activation of protein kinase A (PKA), protein kinase C (PKC) and mitogen-activated protein kinase (MAPK) in cumulus cells are required for gonadotropins-induced meiotic resumption of oocytes. The various actions of cAMP activated by follicle-stimulating hormone (FSH) and luteinizing hormone (LH) also include meiosis activating sterol (MAS), gonadal steroid hormones and epidermal growth factor (EGF) network during meiotic resumption. Another second messenger guanosine 3',5'-cyclic monophosphate (cGMP) induced by nitric oxide (NO) or atrial natriuretic peptide (ANP) also mediates gonadotropins-controlled mammalian oocyte meiotic resumption. The different actions of FSH and LH on meiotic resumption are discussed. We hope to provide a framework to understand how the initial signals generated by gonadotropins-stimulation control the expression of genes required for meiotic resumption. PMID- 17127300 TI - Molecular control of ovulation and luteinization in the primate follicle. AB - In recent years, significant progress was made, particularly through the use of the macaque monkey, in identifying three types of local factors that are induced by the midcycle LH surge and play a critical role in ovulation and/or luteinization of the primate follicle. The ovulatory gonadotropin surge increases prostaglandin (PTG, typically abbreviated PG) levels in follicles prior to rupture; although considerable attention has focused on LH stimulation of the "inducible" form of PG G/H synthase (PTGS2), other aspects of PG synthesis (notably a phospholipase A2, cPLA2, and a PGE synthase, PTGES) and metabolism (15 hydroxy PG dehydrogenase, HPGD) also appear LH-regulated and may control the timing of the PG rise in the ovulatory follicle. Local (intrafollicular) ablation and replacement of PGs suggests that PGE2 is essential for release of the oocyte; but not necessarily for follicle rupture, and not for luteinization. Novel PGE regulated genes are being identified in macaque granulosa cells, including adipose differentiation-related protein (ADFP). Similar types of studies indicate that the rise in progesterone (P) synthesis, as well as the induction of the genomic P receptor in granulosa cells, is essential for both ovulation and luteinization of the primate follicle. Limited data suggest that P action controls cell cycle activity (via cyclin B1 and cyclin-dependent kinase inhibitor p27), cholesterol uptake and utilization (e.g., low density lipoprotein or LDL receptor), proteases and their inhibitors (matrix metalloproteinase or MMP1; tissue inhibitor of MMP or TIMP1) and cell health in the granulosa cell layer. Finally, members of two classes of angiogenic factors, originally proposed as important for embryonic and pathologic (tumorigenic) vasculogenesis, appear induced in the granulosa layer of the preovulatory follicle, i.e., vascular endothelial growth factor (VEGF) and angiopoietin (ANGPT). Local injection of antagonists to VEGF (soluble VEGF receptor) and ANGPT (the natural antagonist ANGPT2) into the preovulatory follicle suppressed ovulation and luteinization in monkeys, possibly by disrupting the structure-function of existing vessels or preventing angiogenesis in the avascular granulosa layer. Further studies using high-throughput genomic and proteomic analysis, particularly on specific cell types (e.g., granulosa, theca and microvascular cells) and distinct follicular regions (apex, base and cumulus-oocyte complex) of the dominant follicle in natural menstrual cycles, are needed. Such information is essential to advance our understanding of the cascade of events leading to ovulation and luteinization of the primate follicle, to unravel the causes of ovary-based infertility and to consider novel ovary-selective approaches to contraception. PMID- 17127301 TI - Magnesium homeostasis in mammalian cells. AB - Mammalian cells tightly regulate cellular Mg2+ content despite undergoing a variety of hormonal and metabolic stimulatory conditions. Evidence from several laboratories indicates that stimulatory conditions that increase cellular cAMP level result in a major mobilization of Mg2+ from cells and tissues into the bloodstream. Conversely, hormones or agents that decrease cAMP level or activate protein kinase C signaling induce a major accumulation of Mg2+ into the tissues. These Mg2+ fluxes are quite large and fast suggesting the operation of powerful transport mechanisms. At front of the recent identification of several Mg2+ entry mechanisms, the Mg2+ extrusion pathway(s) still remain(s) poorly characterized. Similarly, it remains not completely elucidated the physiological significance of these Mg2+ fluxes in the various tissues in which they occur. In the present review, we will attempt to provide a comprehensive framework of the modalities by which cellular Mg2+ homeostasis and transport are regulated, as well as examples of cellular functions regulated by changes in cellular Mg2+ level. PMID- 17127302 TI - Glutamine in the central nervous system: function and dysfunction. AB - Glutamine (Gln) abounds in the central nervous system (CNS), and its interstitial and cerebrospinal fluid (CSF) concentrations are at least one order of magnitude higher than of any other amino acid. Gln transport from blood to the brain is insufficient to meet the demand of the brain tissues for this amino acid. This demand is met by intracerebral Gln synthesis from glutamate (Glu), a reaction carried out by glutamine synthetase (GS), an enzyme residing in astrocytes. A major proportion of astroglia-derived Gln is shuttled to neurons where it is degraded by phosphate-activated glutaminase (PAG) giving rise to the excitatory neurotransmitter amino acid Glu, which is also a precursor of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). Glu released from neurons is taken up by astrocytes, and reconverted to Gln, closing the so called "glutamate glutamine" cycle. A portion of Gln serves as an energy metabolite, and part of it leaves the brain to blood. Gln efflux from astrocytes, its neuronal uptake and egress to the blood via the cerebral capillary endothelial cells is mediated by different amino acid carriers showing i) considerable preference for Gln, ii) distribution between astrocytes and neurons that favors astrocyte-to-neuron fluxes of the amino acid. The Gln-specific carriers also largely contribute to Gln efflux from the brain to the vascular bed. Excessive accumulation of Gln in brain cells may be deleterious to brain function. In hyperammonemia associated with acute liver failure, excess Gln leads to cerebral edema, which largely results from its interference with mitochondrial function and partly from its osmotic action. Future analyses of the roles of Gln in both normal and abnormal cerebral metabolism and function will have to account for its newly recognized direct involvement in the regulation of gene transcription and/or translation. PMID- 17127303 TI - Glutamine, gene expression, and cell function. AB - Glutamine is the most abundant free amino acid in the body and is known to play a regulatory role at the gene and protein level in several cell specific processes including metabolism (e.g. oxidative fuel, gluconeogenic precursor and lipogenic precursor), cell integrity (survival, cell proliferation), protein synthesis and degradation, redox potential, respiratory burst, insulin resistance, insulin secretion and extracellular matrix synthesis. Glutamine has been shown to regulate the expression of many genes related to metabolism, signal transduction, cell defense and repair and to activate intracellular signaling pathways. Thus, the function of glutamine goes beyond that of a simple metabolic fuel or protein precursor as previously assumed. In this review, we have attempted to identify some of the common mechanisms underlying glutamine dependent changes in gene and protein expression and cellular function. PMID- 17127304 TI - Glutamine signalling in bacteria. AB - Glutamine is a metabolite of central importance in bacterial physiology. In addition to its function as one of the 20 standard amino acids in protein synthesis, glutamine is required for the biosynthesis of a variety of nitrogen containing compounds. Of particular importance is glutamine synthesis as primary reaction of ammonium assimilation. Because of this versatile role, glutamine metabolism is tightly controlled in response to the cellular nitrogen status in bacteria. Recent progress in elucidating the molecular basis of nitrogen signalling has shed light on the role of glutamine as a signalling molecule. Bacteria belonging to the phylogenetic domains of proteobacteria and low G+C gram positives (firmicutes) have evolved different mechanisms to monitor glutamine as an indicator of the state of nitrogen metabolism, which then regulates nitrogen metabolism at the transcriptional and post-transcriptional levels. Using the conserved PII signal transduction system, major groups of prokaryotes, including the cyanobacteria, have evolved yet another strategy to monitor the cellular nitrogen status, which relies on 2-oxoglutarate instead of glutamine as the signalling molecule. In addition to monitoring the intracellular glutamine level, bacteria may respond to extracellular glutamine, which is used as a nutrient. This overview details our current knowledge of glutamine-regulated processes in bacteria. PMID- 17127305 TI - Glutamine metabolism and signaling in the liver. AB - Glutamine is the most abundant amino acid in the human body and can be synthesized by almost all tissues by the glutamine synthetase (GS)-catalyzed amidation of glutamate. Hepatocytes have access to extracellular glutamine by the concentrative uptake via members of the sodium-dependent neutral amino acid transport systems N and A. Hepatic glutamine metabolism in connection with urea synthesis is importantly involved in systemic ammonia detoxication and pH regulation due to the unique regulatory properties of the liver-type glutaminase, the acinar compartimentation of urea and glutamine synthesis, and a cycling of glutamine between periportal and perivenous hepatocytes. Upregulation of GS expression in hepatocellular carcinoma is related to growth advantage and an enhanced metastatic potential. Glutamine is a potent activator of signal transduction. Recent progress concerns the understanding of glutamine-induced hepatocyte swelling and the downstream activation of integrins, Src, and MAP kinases in the regulation of autophagic proteolysis, canalicular bile acid excretion, glycogen and fatty acid synthesis, insulin signaling, and protection from apoptosis. Most recently the first primary GS defect leading to inherited glutamine deficiency with fatal outcome was described in human. This review summarizes recent progress in the understanding of glutamine metabolism and signal transduction, which provides further rationale for the use of glutamine as a therapeutic tool. PMID- 17127306 TI - Quality of life in oncology with emphasis upon neuro-oncology. AB - Quality of life, as a science has been steadily gaining importance in both clinical practice as well as research. Despite major progress in the development of validated and clinically-relevant health-related quality of life (HRQOL) measures, we still face many challenges in bridging the gap between what we know and how to apply it in clinical practice: in making the transfer from the mere collection of QOL data to its utilization in improving patient outcome through interventional symptomatic therapy. This manuscript traces the development of QOL as a science to its potential utility in both clinical care and clinical research, as well as an outcomes measure. The emphasis has been placed upon quality of life in oncology with special attention to neuro-oncology. PMID- 17127307 TI - Signaling mechanisms utilized by antigen receptors and integrins: common intermediates, different outcomes? AB - Antigen receptors and integrins are structurally and functionally distinct, but both play key roles in regulating immune cell activation and function. Understanding the molecular basis of the signaling pathways utilized by antigen receptors and integrins is fundamental to identifying the mechanisms underlying immune system function and dysfunction (e.g. autoimmune disease) and identifying potential targets for modifying the immune response with therapy. Recently, several key regulators of antigen receptor signaling have also been revealed to be important molecular intermediates in integrin-triggered signaling pathways. These include the protein tyrosine kinase Syk, the guanine nucleotide exchange factor Vav, and the adaptor protein SLP-76. While antigen-receptor signaling is generally associated with leukocyte activation and differentiation, integrins are most commonly thought of as adhesive receptors. This raises the interesting question of how common molecular intermediates may regulate diverse cellular processes such as activation versus adhesion and migration, and provides a framework for defining potentially unique mechanisms utilized by cells of the immune system to regulate integrin-dependent cell function. PMID- 17127308 TI - The role of actomyosin and the microtubular network in both the immunological synapse and T cell activation. AB - The formation of immunological synapses between T cells and APCs, as well as the functions associated with this structure, like cytokine secretion and the lysis of infected cells, are critical processes in the immune response. The T cell cytoskeleton is involved in these activities and this has been the subject of numerous studies in recent years. Although the importance of the T cell actin network is evident, the participation of microtubules and myosin motors in the formation of immunological synapses and T cell effector functions has also been assessed. This review provides an update on the role of cytoskeletal networks and related molecules in the activation of T lymphocytes. PMID- 17127309 TI - WNTS and WNT receptors as therapeutic tools and targets in human disease processes. AB - The body of scientific literature linking Wnts and Wnt-associated proteins to human disease processes continues to grow in parallel with new discoveries from basic science laboratories that further characterize the elaborate cellular events following the binding of Wnts to their receptors. While Wnt-mediated signaling has long been known to play a major role in human carcinogenesis, accumulating evidence indicates that Wnts are also important mediators of inflammation and recovery from injury. The binding of secreted Wnt ligands to their receptors offers an attractive and accessible target for therapeutic regulation of these signaling pathways. Several promising preliminary studies have already addressed potential avenues for the manipulation of Wnt signaling in disease processes. This review will focus on disease processes involving the regulation of Wnt signaling at the level of Wnt binding to its target receptors. Wnt proteins, Wnt receptors, and secreted Wnt inhibitors are attractive as potential therapeutic agents and targets due to their extracellular location. In addition, since Wnt signaling results in a diverse array of downstream intracellular events, many of which are not fully understood, the targeting of this pathway at the most upstream site of pathway activation also provides a strategic advantage for therapy. As the list of Wnt-related diseases continues to grow, advances in our understanding of the biochemical and molecular mechanisms underlying Wnt signaling may ultimately translate into innovative ways to treat Wnt-related disease processes in patients. PMID- 17127310 TI - Wnt/FZD signaling and colorectal cancer morphogenesis. AB - Malignant progression of colorectal carcinomas is characterized by an epithelial mesenchymal transition (EMT)-like de-differentiation of the invading tumor cells. However a re-differentiation towards an epithelial phenotype, resembling a mesenchymal-epithelial transition (MET), is detectable in metastases. This indicates that malignant progression is based on dynamic processes, which can not be explained solely by irreversible genetic alterations, but must be additionally regulated by the tumor environment. The main oncoprotein in colorectal cancer is the Wnt-pathway effector beta-catenin, which in most cases is overexpressed due to mutations in the adenomatous polyposis coli (APC) tumor suppressor. EMT of tumor cells is associated with a nuclear accumulation of the transcriptional activator beta-catenin, which is reversed in metastases. Nuclear beta-catenin is involved in two fundamental processes in embryonic development: EMT and stem cell formation. Accumulating data demonstrate that aberrant nuclear expression of beta catenin can also confer these two abilities to tumor cells, indicating the crucial role of aberrant Wnt-signaling for malignant tumor progression. PMID- 17127311 TI - WNT signaling in the normal intestine and colorectal cancer. AB - The intestinal epithelium is a self-renewing tissue that represents a unique model for studying interconnected cellular processes such as proliferation, differentiation, cell migration and carcinogenesis. This review covers work from the past decade and highlights the importance of the canonical Wnt pathway in regulating multiple aspects of intestinal homeostasis. Numerous in vivo studies combined with gene profiling experiments have shown that Wnt signaling promotes maintenance of epithelial stem cells and early progenitors by driving transcription of genes associated with proliferation. These studies also revealed strong similarities between the genetic program initiated by Wnt signals in normal crypt progenitors and in colorectal cancer cells. More recently it has become apparent that Wnts do not act alone but rather cooperate with Notch signals in maintaining progenitor cell populations. Processes associated with differentiated epithelial cells also appear to be regulated by Wnt signals. For instance, Paneth cells employ active Wnt signals for terminal differentiation. Moreover, through transcriptional regulation of members of the Eph and Ephrin families, Wnt signaling promotes compartmentalization of epithelial cells along the crypt-villus axis. The Eph/Ephrin system also operates to limit progression of colorectal cancer beyond the early stages. PMID- 17127312 TI - Wnt-Frizzled signalling and the many paths to neural development and adult brain homeostasis. AB - The regulation of brain development and function is the result of complex cell restricted and temporal expression profiles directed by signaling networks constantly imposing exquisite regulatory control on many genes at any one moment within a cell. The ultimate outcome is a genetically controlled balancing act where expression profiles of these hundreds of genes result in cellular proliferation, differentiation and the ultimate choice between long-term survival and apoptosis. During embryonic development there is a massive expansion of neurons and glia, which is balanced with programmed cell death as the brain matures and remodels. As developing brain cells differentiate, they migrate toward the region where they will ultimately seek out interactions with other cells and perform their specialized tasks. Although a number of signaling pathways have been shown to contribute to various processes allowing the maintenance of normal neurogenesis, the precise signaling machinery necessary for modulating the maintenance of both the neuroblast and differentiated neuronal population, and regulating transition between the two, is still being solved. Not surprisingly, the Wnt signaling pathway is important in regulating neural development but also appears to be involved in adult neurogenesis and some brain disorders. Here, we review key findings showing the pivotal nature of Wnt Frizzled (FZD) signaling in neurogenesis as revealed by a number of molecular genetic studies using mice and other model organisms. We also review the current literature on the role of the Wnt pathway in the generation of brain cancers, particularly the most common primitive neuroectodermal tumors in childhood, neuroblastomas, and in neurodegenerative diseases such as Alzheimer's disease. PMID- 17127313 TI - Mechanical regulation of matrix metalloproteinases. AB - Matrix metalloproteinases can degrade and modify almost all components of the extracellular matrix hence their enzymatic activity is tightly regulated under physiological conditions. Primary modes of enzyme regulation include transcriptional control, zymogen activation and dynamic inhibition by tissue inhibitors of matrix metalloproteinases. Recent studies have demonstrated that mechanical regulation of matrix metalloproteinases largely operate through these regulatory pathways. Over the last decade a large cohort of studies have been conducted on many tissue/cell types using diverse loading parameters in vivo and in vitro suggesting that mechanical load is essential in maintaining normal tissue function via the matrix metalloproteinases. However there may be a mechanically-regulated homeostasis, with cells responding to and interpreting growth factors and other biochemical signals within the context of mechanical forces to provide a suitable cellular matrix metalloproteinase response. On the contrary, mechanical overload can result in unrestrained matrix metalloproteinase activities eventually leading to matrix degradation, mechanical dysfunction and failure of the tissue. In this chapter, the effect of mechanical load on matrix metalloproteinase expression will be reviewed, and the signal transduction pathways involved in modulating the metabolic homeostasis of various tissues including blood vessels, intervertebral disc and components of the synovial joint with emphasis on articular cartilage discussed. Both mechanically-induced stimulation and inhibition of matrix metalloproteinases will be discussed and placed into context with their potential relevance to disease. PMID- 17127314 TI - Acetylation in the regulation of metalloproteinase and tissue inhibitor of metalloproteinases gene expression. AB - Together, the matrix metalloproteinases (MMPs) are capable of degrading every component of the extracellular matrix (ECM). Besides degradation of the ECM, MMPs release bioactive molecules from the matrix or cell surface and play important role in tissue repair after injury, development and in a number of pathologies including arthritis and cancer metastasis. Small molecules that inhibit a broad spectrum of metalloproteinases have not proved useful in the treatment of various diseases, probably due to the diverse roles of this large family of enzymes. An alternative therapeutic approach for a number of pathologies is to modulate the expression of specific metalloproteinase genes. Acetylation represents a recently identified covalent protein modification that is strongly implicated in transcriptional regulation. Histones were the first proteins demonstrated to show variable acetylation leading to gene activation. Subsequently, a large number of molecules including structural proteins, intracellular signaling molecules, nuclear membrane receptors and transcription factors were shown to be acetylated. Acetylation, like phosphorylation, is a reversible modification. Acetyl groups are added by a family of histone acetyl transferase enzymes (HATs) and are removed by histone deacetylases (HDACs). Inhibitors of HDACs (HDACi) have potent anti-proliferative and pro-apoptotic activities in cancer cells and may be used as cancer therapeutics. In this review, we examine the impact of changes in acetylation on the expression of the MMPs and their inhibitors (tissue inhibitors of metalloproteinases, TIMPs). We discuss the suggestion that HDACi may act in a dual fashion: selectively decreasing cancer cell viability and reducing metastatic potential by decreasing stromal cell expression of specific metalloproteinases. Furthermore, we consider the possibility that selective HDACi have a potential as anti-inflammatory agents and in a range of degradative diseases such as arthritis. PMID- 17127315 TI - Collagenase gene regulation by pro-inflammatory cytokines in cartilage. AB - The essentially irreversible degradation of articular cartilage collagen represents a key, rate-limiting process in arthritic diseases. This process is typically initiated as a consequence of an inflammatory response, and if left unchecked ultimately leads to loss of joint function, pain, disability and a need for joint replacement surgery. Although we have identified the enzymes capable of effecting such destructive proteolysis, and considerable evidence indicates that tumour necrosis factor alpha and interleukin-1 are major pro-inflammatory mediators in joint destruction, we still know relatively little about how these mediators regulate collagenase gene expression in chondrocytes. Inflammatory arthritis has long been considered to be synovium-driven but compelling data now also implicate the chondrocyte, the sole cell type present in cartilage, as an active player in the destructive process. An understanding of how different cytokines interact, and how the pathways they activate cross-talk will not only provide important new insight into the mechanisms of joint destruction but also identify new targets for therapeutic intervention. PMID- 17127316 TI - Corticotropin releasing hormone: a diagnostic marker for behavioral and reproductive disorders? AB - Corticotropin-releasing hormone (CRH) is a key mediator of endocrine, autonomic, behavioral, and immune responses to stress. The ability of CRH to induce hormonal stress responses has been used to investigate the functionality of the hypothalamus-pituitary-adrenal axis, and consequently the activity of hypothalamic CRH neuronal systems. Indeed, CRH administration to humans causes prompt release of ACTH, followed by secretion of cortisol, aldosterone and other adrenal steroids. CRH hypersecretion/hyperactivity has been associated to major depression, anxiety-related disorders, anorexia nervosa, Alzheimer's and Parkinson's diseases and progressive supranuclear palsy. During pregnancy the human placenta and its accessory membranes are the major sites of CRH synthesis and secretion. Placental CRH secretion is autonomous, but increasing evidence indicates that maternal or fetal conditions may influence such secretion. Therefore, the emerging concept is that in the event of acute or chronic metabolic, physical or infectious stress, the placenta takes part in a stress syndrome by releasing CRH, which may contribute to restore local blood flow and to influence the timing of delivery. The CRH released by the placenta is measurable in maternal plasma and other biological fluids and may be used to diagnose subclinical processes anteceding pregnancy complications such as pre eclampsia and preterm delivery. PMID- 17127317 TI - The role of CRH receptors and their agonists in myometrial contractility and quiescence during pregnancy and labour. AB - The mechanism of human labor remains a scientific enigma. Corticotropin-releasing hormone (CRH), a hypothalamic peptide that controls the response of the body to stress and which is also produced by the placenta and intrauterine tissues during pregnancy is potentially involved in the onset of labor. CRH is part of a family of mammalian peptides that includes the urocortins (UCNs), which are also expressed by the placenta and intrauterine tissues. During human pregnancy, CRH appears to target multiple feto-maternal tissues, including the myometrium, implicating CRH in the regulation of the transition from relaxation to active uterine contractions. The myometrial actions of CRH are mediated via a wide network of specific G-protein coupled membrane-bound receptors. These receptors have various functional properties, depending on the receptor subtype, the ability of agonists to activate specific signalling cascades and the stage of pregnancy. In addition, their function is dependent upon other intracellular signals via communication between signalling cascades, suggesting potential multiple roles of CRH and other CRH-like peptides during pregnancy and labor. This review will provide the current concepts about the role of CRH and UCNs and their myometrial receptors during pregnancy, labor and delivery. PMID- 17127318 TI - Peripheral corticotropin-releasing hormone is produced in the immune and reproductive systems: actions, potential roles and clinical implications. AB - Corticotropin-releasing hormone (CRH), the principal regulator of the hypothalamic-pituitary-adrenal axis, has been identified in various organ systems, including the immune and the female and male reproductive systems. CRH like immunoreactivity has been reported in peripheral inflammatory sites and in a number of reproductive organs, including the ovaries, endometrial glands, decidualized endometrial stroma, placenta, decidua, and the testes. Therefore, "immune" and "reproductive" CRH are forms of "tissue" CRH; i.e., CRH found in peripheral tissues. Immune CRH plays a direct immunomodulatory role as an autocrine/paracrine mediator of inflammation. Immune CRH participates in several experimental inflammations and, in humans, in inflamed tissues from patients with autoimmune and inflammatory diseases. One of the early effects of immune CRH is the degranulation of mast cells and the release of histamine and several inflammatory cytokines. Reproductive CRH is regulating reproductive functions with an inflammatory component, such as ovulation, luteolysis, decidualization, implantation, and early maternal tolerance. Placental CRH participates in the physiology of pregnancy and the onset of labor. Circulating placental CRH is responsible for the physiologic hypercortisolism of the latter half of pregnancy. Postpartum, this hypercortisolism is followed by a transient adrenal suppression, which may explain the blues/depression and increased autoimmune phenomena observed during this period. PMID- 17127319 TI - Advances in understanding corticotrophin-releasing hormone gene expression. AB - Glucocorticoids inhibit corticotrophin-releasing hormone (CRH) gene expression in the hypothalamic paraventricular nucleus (PVN), but stimulate expression in the placenta. In AtT20 cells (a model of PVN CRH production) cAMP produces a high level of promoter activity. Cyclic AMP stimulation occurs through the cAMP response element (CRE) and the caudal type homeobox protein response element (CDXARE). The CRE acts as part of a cAMP response unit that includes the hybrid steroid response element (HRE), ecdysone response element (EcRE), metal responsive transcription factor-1 response element (MTFRE), ying yang 1 response element (YY1RE) and negative glucocorticoid response element (nGRE). Cyclic AMP acts on the HRE, EcRE and MTFRE to block YY1RE mediated inhibition of the CRE. Glucocorticoids acting at the nGRE inhibit cAMP activation of the CRE. In placental cells the CRH promoter has low intrinsic basal activity and cAMP causes a modest increase in activity. Stimulation by glucocorticoids and cAMP and inhibition by estrogen and estrogen receptor alpha occurs through the CRE. In AtT20 cells multiple response elements coordinate a response to cAMP and glucocorticoids while in placental cells the CRE acts in isolation. These differences in promoter function lead to responses that meet specific physiological needs. PMID- 17127320 TI - Activation of mitogen-activated protein kinase pathways by the granulocyte colony stimulating factor receptor: mechanisms and functional consequences. AB - The cytokine Granulocyte Colony Stimulating Factor (G-CSF) promotes proliferation, differentiation, survival and functional maturation of cells within the neutrophilic granulocyte lineage. G-CSF binds to its cell-surface receptor (G-CSFR) causing activation via homodimerisation and subsequent phosphorylation on four tyrosine residues of the receptor intracellular domain. This initiates a range of intracellular signalling events including the activation of Mitogen-Activated Protein Kinase (MAPK) pathways. G-CSF stimulates activation of the ERK 1/2 pathway, as well as the stress-activated JNK and p38 pathways, and the less-characterised ERK5/Big MAPK 1 pathway. Receptor mutagenesis studies have aided in the identification of regions of the G-CSFR that mediate specific activation of these MAPK pathways. In addition, the activation of individual MAPK pathways appears to contribute to distinct biological outcomes. Thus, MAPK activation may be an important mediator of the actions of G-CSF. PMID- 17127321 TI - Granulocyte colony-stimulating factor and its receptor in normal myeloid cell development, leukemia and related blood cell disorders. AB - Granulocyte colony-stimulating factor (G-CSF) is the major hematopoietic cytokine involved in the control of neutrophil production and thus serves as a critical regulator of the innate immunity against bacterial infections. G-CSF is applied on a routine basis in the clinic for treatment of congenital and acquired neutropenias, diseases characterized by a critical shortage of neutrophils, leading to severe opportunistic bacterial infections. Very recently, it has become clear that therapeutic application of G-CSF may not be limited to different types of neutropenia, but may extend to non-hematological conditions, in particular cardiac and brain infarctions. G-CSF drives the proliferation, survival and neutrophilic differentiation of myeloid progenitor cells by activation of a receptor of the hematopoietin receptor superfamily, which subsequently triggers multiple signaling mechanisms. These mechanisms exert positive as well as negative effects on the signaling function of the G-CSF receptor. The integrated output of these signaling pathways provide the appropriate balance needed for accurate production of neutrophils under both steady state and "emergency" conditions. Here we review how these mechanisms are thought to act in concert to meet with these demands and how perturbations in the function of the G-CSF receptor are implicated in various types of myeloid disease. PMID- 17127322 TI - The role of the granulocyte colony-stimulating factor receptor (G-CSF-R) in disease. AB - Granulocyte colony-stimulating factor (G-CSF) is a key regulator of granulopoiesis via stimulation of a specific cell-surface receptor, the G-CSF-R, found on hematopoietic progenitor cells as well as neutrophilic granulocytes. It is perhaps not surprising, therefore, that mutations of the G-CSF-R has been implicated in several clinical settings that affect granulocytic differentiation, particularly severe congenital neutropenia, myelodysplastic syndrome and acute myeloid leukemia. However, other studies suggest that signalling via the G-CSF-R is also involved in a range of other malignancies. This review focuses on the molecular mechanisms through which the G-CSF-R contributes to disease. PMID- 17127323 TI - Oxytocin and parturition: a role for increased myometrial calcium and calcium sensitization? AB - Preterm birth is associated with the majority of all death and chronic disability related to pregnancy, birth and the neonatal period. The costs to families and to the health care system are enormous. Current approaches to prevent or arrest preterm labour have been unsuccessful. This failure is largely based on our poor understanding of the regulation of the timing and maintenance of parturition. Oxytocin (OT) is the most potent known uterine stimulant. It is produced in the hypothalamus and secreted into the maternal bloodstream. However, OT also is produced within the uterine decidua in late gestation and the concentrations increase around the time of labour onset. The receptor for OT (OTR) is a G protein coupled receptor linked through G alpha(q/11) to phospholipase C (PLC). Activation of PLC causes increased inositol trisphosphate (IP3) and diacyl glycerol (DAG). IP3 activates specific receptors in the sarcoplasmic reticulum to release Ca2+ into the cytosol. This may induce further influx of Ca2+ from the extracellular space and the increased Ca2+, after binding to calmodulin, activates myosin light chain kinase to phosphorylate myosin light chains (MLC) and cause contraction of the myocyte. DAG activates protein kinase C (PKC), several isoforms of which have been implicated in uterine contraction, but the substrates for this enzyme in the uterine myocyte are essentially unknown. Oxytocin may also cause "Ca2+-sensitization," a process whereby there is a greater contractile force generated from a given increase in cytosolic Ca2+, although the contribution of this process to myometrial contraction remains an area of debate. This phenomenon occurs mainly due to inhibition of myosin light chain phosphatase (MLCP), the enzyme that reverses the phosphorylation of MLC. There are several important potential mediators of this MLCP-inhibitory pathway in the myometrium, including the small monomeric G-protein RhoA, its downstream kinase Rho-associated kinase (ROK). and the 17-kDa PKC-potentiated inhibitor of protein phosphatase 1c (CPI-17). The roles in the myometrium of other recently identified MLCP interacting molecules also requires further investigation. These Ca2+-sensitization pathways could be important in the mechanisms underlying pre term or term labour. An increased understanding of the complexities of the multitude of regulatory mechanisms for uterine contractility may lead to new pharmacologic agents for the prevention or reversal of uterine contractions. This, in turn, is necessary to facilitate the development of novel and effective strategies to reduce the incidence of preterm birth. PMID- 17127324 TI - Progesterone receptor profile in the decidua and fetal membrane. AB - Whereas in most mammals the onset of labor is preceded by a rapid fall in the maternal progesterone levels, in humans and in higher primates, maternal, fetal and amniotic fluid concentrations of progesterone are sustained before the onset of labor. Therefore, the mechanism for parturition, which has been proposed for humans, is 'functional' progesterone withdrawal. This review is focused on the expression profile, activity and interaction of the progesterone receptor (PR) isoforms in the decidua and the fetal membrane during the initiation of labor. Binding of progesterone to PR induces a significant conformational change on the receptor proteins. These changes result in dimerization, increased receptor phosphorylation and binding of receptor dimers to specific hormone responsive DNA elements in the promoter of target genes. Interaction with specific co-activator proteins and general transcription factors are responsible for the formation of a productive transcription initiation complex. The PR also mediates the activation of cytoplasmic signaling pathways, participating in the induction of signal transduction pathway in the cytoplasm. Balanced expression of the two major progesterone receptors isoforms is crucial for progesterone function as uterine muscle inhibitor. Change in PR isoforms profile seems to be responsible for decidual activation. Decidua without contractions shows consistent profile with PR-B being the dominant isoform. PR-A, PR-C and two additional truncated isoforms are also detected but in significantly smaller concentration. After initiation of contractions, a sharp decline in PR-B shifts the PR-A/PR-B ratio toward PR-A dominance. This shift in the decidua towards increase expression of progesterone receptor isoform A and decrease in PR isoform B is having a pivotal role in decidual activation and initiation of labor. PMID- 17127325 TI - The matrix metalloproteinases (MMPS) in the decidua and fetal membranes. AB - The role of the matrix metalloproteinases (MMPs) in the decidua, fetal membranes and amniotic fluid (AF) has been receiving more and more attention. The MMPs are not only important intermediaries in pathological processes leading to preterm labor but it seems that they also play a crucial role in the activation of labor at term. During normal gestation MMP-1, -2, -3, -7 and -9 are found in the amniotic fluid and fetal membranes. MMP-2 and MMP-3 are expressed constitutively while MMP-9 is barely detectable until labor. At labor, while MMP-9 is the major MMP responsible for gelatinolytic activity in the membranes, MMP-2 is dominant in the decidua. MMP-7 (AF) increases with gestation but does not appear to play a major role in labor. The expression of MMPs is attenuated through the expression of relaxins, integrins and extracellular matrix metalloproteinase inducer (EMMPRIN). Spontaneous preterm delivery (PTD) may be a product of preterm labor (PTL), preterm premature rupture of membranes (P-PROM) or placental abruption. Each of these processes may have differing pathways but the presence of an intrinsic inflammatory response with or without infection seems to involve all etiologies. The inflammatory response is mediated with cytokines such as interleukins -1, -6 and -8 and tumor necrosis factor alpha. MMP-3, MMP-7 and MMP 8 appear to be important in these processes. MMP-9, which is the major MMP involved in normal labor, plays an important role in pathological labor as well. Finally, apoptosis seems to play a role in pathological labor, particularly deliveries involving P-PROM. African-American are at greater risk of PTD than white or Hispanic Americans. Environmental differences may not suffice to explain this phenomenon. Genetic polymorphisms of the MMP genes may help explain the greater risk among this population. Finally, manipulating MMPs may have a role in the prevention of PTD. Agents suggested include indomethacin, N-acetylcysteine, progesterone and specific inhibitors of phosphodiesterase 4. PMID- 17127326 TI - Persistent donor-specific alloreactivity may portend delayed liver rejection during drug minimization in children. AB - Immunoreactivity, immunosuppression requirement and liver graft function was assessed serially for its relationship to delayed/recurrent acute cellular rejection (ACR) after the first 60 days in 36 pediatric primary liver transplant (LTx) recipients. Subjects were classified as rejectors (n=20) or Non-Rejectors (n=16) based on the presence/absence of biopsy-proven ACR in the first 60 days. All children received anti-lymphocyte induction and steroid-free Tacrolimus or Sirolimus monotherapy, as reported previously. Median age was 4 years (0.45-18) and follow-up was 570 days (106-1144). Compared with non-rejectors, rejectors 1. took significantly longer to achieve reduced donor-specific alloreactivity by MLR (p=0.049), and "low" TAC/SRL whole blood requirements defined as TAC levels < or = 8 ng/ml (p=0.0048), 2. experienced significantly greater variation in time to achieve reduced donor-specific immunoreactivity (SEM 0.8 vs 3.85, p=0.0048), and 3. experienced greater ACR incidence during minimization of immunosuppression (35% versus 6%, p=0.032). Serial monitoring of immunoreactivity may increase the safety with which immunosuppression is minimized in pediatric LTx. PMID- 17127327 TI - Applications of flow cytometry to mycoplasmology. AB - Flow cytometry has become a valuable tool in different fields of microbiology, such as clinical microbiology, aquatic and environmental microbiology, food microbiology, and biotechnology. It combines direct and rapid assays to determine numbers, biochemical and physiological characteristics of individual cells, revealing the heterogeneity present in a population. This review focuses on the applications of flow cytometry to the field of mycoplasmology. It tries to give a scope of the important breakthroughs which occurred in this field in the last decades, and in the advantages of introducing flow cytometry in research and routine diagnostic procedures of mycoplasmas. PMID- 17127328 TI - Phytoplasmas: diversity, taxonomy, and epidemiology. AB - Phytoplasma associated diseases are spread worldwide, and in several cases are associated with severe epidemic of very often quarantine importance. These plant pathogens are prokaryotes belonging to the Mollicutes class since they lack a cell wall; up to now they were not cultivated in axenic culture therefore Koch postulates are only sometimes fulfilled by using alternative tools, such as graft or insect transmission. The possibility to design specific primers for highly conserved genes such as 16S ribosomal gene together with the use of molecular probes randomly cloned from phytoplasma genome, allowed discriminating and molecularly classifying them. Now a certain amount of knowledge is available that allow starting epidemiological studies in order to prevent further spreading of phytoplasma-associated diseases. In this paper molecular, biological and epidemiological characteristics of phytoplasma associated with important diseases worldwide are described. PMID- 17127329 TI - Adhesion proteins of Mycoplasma pneumoniae. AB - Mycoplasmas are unique cell wall deficient, cholesterol requiring, highly pleomorphic bacteria that possess very small genome. M. pneumoniae is an important human pathogen that possesses specialized tip organelle to mediate cytadherence. It is a complex multifactorial process requiring a group of mycoplasma proteins such as P1, P30, P116 and HMW1-3. Expression of major mycoplasma adhesin proteins in heterologous expression systems provides an opportunity to study the role(s) of these proteins in pathogenicity and helps to develop diagnostic reagents. In this review we highlight the role(s) of various cytadhesin proteins of M. pneumoniae. PMID- 17127330 TI - Ubiquitin-like protein modifications in prostate and breast cancer. AB - Post-translational modifications by ubiquitin-like proteins have been implicated in the regulation of diverse cellular processes, including nuclear transport, transcription regulation, stress response and DNA repair. Ubiquitination is well characterized for its roles in regulating these cellular processes. As a newly identified member of ubiquitin-like proteins, the small ubiquitin-like modifier (SUMO) has received a great deal of attention for its functions distinct from ubiquitin. In particular, alterations of SUMO conjugation or sumoylation have been implicated in several human diseases, including cancer. Although little is known about the underlying mechanism of sumoylation-associated tumorigenesis, the modulation of nuclear receptor (NR)-mediated signaling pathways is likely to play a role in this aspect. NRs are a family of ligand dependent transcription factors which control cell growth and differentiation in many cell types, as well as during the development of cancer. In this review, we will discuss some basic aspects of sumoylation and how sumoylation modulates the NR-mediated gene expression, focusing on androgen receptor (AR) and estrogen receptor (ER), a key player in progression of prostate or breast cancer. PMID- 17127331 TI - Neuroprotective effect of granulocyte-colony stimulating factor. AB - Granulocyte-colony stimulating factor (G-CSF) is a growth factor which stimulates proliferation, differentiation, and survival of hematopoietic progenitor cells. G CSF is being used extensively in clinical practice to accelerate recovery of patients from neutropenia after cytotoxic therapy. However, growing evidences have suggested that G-CSF has important non-hematopoietic functions in central nervous system. Recent studies have shown the presence of G-CSF/G-CSF-receptor (G CSFR) system in the brain, and their roles in neuroprotection and neural tissue repair as well as improvement in functional recovery. The increased expression of G-CSF/G-CSFR on neurons subjected to hypoxia provides evidence that G-CSF may have an autocrine protective signaling mechanism in response to neural injury. G CSF exerts neuroprotective actions through the inhibition of apoptosis and inflammation and the stimulation of neurogenesis. Moreover, G-CSF has been shown to mobilize bone marrow stem cells into the injured brain improving neural plasticity. In this review, we summarize some of the recent studies on G-CSF and the corresponding signal transduction pathways regulated by G-CSF in neuroprotection. PMID- 17127332 TI - Therapeutic hypothermia: neuroprotective mechanisms. AB - Hypothermia has long been known to be a potent putative neuroprotectant. Experimental evidence and clinical experience show that hypothermia protects the brain from cerebral injury. Recent insights into the mechanisms of cerebral ischemia and reperfusion suggest reasons why hypothermia may be an ideal modality for stroke therapy. Hypothermia protects brain tissue in multiple ways. It retards energy depletion, reduces intracellular acidosis, lessens the ischemia related accumulation of excitotoxic neurotransmitters, and attenuates the influx of intracellular calcium. Additionally, hypothermia suppresses the generation of oxygen free radicals involved in secondary damage associated with reperfusion. It also suppresses the mechanisms related to blood-brain barrier degeneration and postischemic remodeling. The clinical application of therapeutic hypothermia and its limitations will be summarized in this paper. Therapeutic hypothermia is likely to undergo phase III clinical trials in various clinical settings. Novel technologies are being developed to optimize the safety and efficacy of this promising approach. PMID- 17127333 TI - Dodging the CTL response: viral evasion of Fas and granzyme induced apoptosis. AB - The importance of CTL induced apoptosis as a vital part of the protection of host organisms from pathogenic viruses cannot be overstated. Conversely, the ability of a virus to evade CTL induced apoptosis is equally important to its survival. Important insights in viral pathogenesis and host immunology have been discovered through observations of this constantly evolving interchange. This mini review will build upon previously published comprehensive reviews by reorganizing the anti-apoptotic strategies specific for CTL induced apoptosis and integrating recent discoveries in viral evasion of Fas/FasL and perforin/granzyme mediated apoptosis. This updated look at viral evasion in the context of the CTL response should generate dialogue and provide impetus for research to illuminate interactions between the best defense against viruses and the viral adaptations to evade this defense. PMID- 17127334 TI - The role of tau phosphorylation and cleavage in neuronal cell death. AB - The microtubule-associated protein tau is the primary component of the intracellular filamentous deposits found in Alzheimer's disease (AD) brain and also in a family of neurodegenerative diseases called 'tauopathies', where tau pathology is the primary, defining characteristic with little or no amyloid-beta (Abeta) pathology. It has been demonstrated that tau modifications such as hyperphosphorylation and truncation might be important events in the process leading to tau intracellular aggregation and neuronal cell death. The discovery of tau gene mutations in frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17) reinforced the predominant role attributed to tau proteins in the pathogenesis of neurodegenerative disorders. This review highlights recent findings concerning the normal metabolism and function of tau, as well as the abnormal processing and function of tau in AD and in the tauopathies. PMID- 17127335 TI - Cetuximab is an active treatment of metastatic and chemorefractory thymoma. AB - Advanced chemorefractory thymic epithelial tumors still represent a challenge in clinical oncology. A rationale-based therapeutic approach targeting a key pathway should represent the ideal solution in a neoplasm that can over-express Epidermal Growth Factor Receptor (EGFR) in the epithelial component. On the basis of these considerations, two patients with metastatic heavily pretreated disease were evaluated for EGFR expression in the primitive tumor, being considered this data as a basis for an anti EGFR treatment with the monoclonal antibody cetuximab which targets EGFR. A strong EGFR expression was revealed by immunohistochemistry in the two cases considered, thus the patients received cetuximab and reported a partial response as assessed by Computed Tomography (CT), Positron Emission Tomography (PET) and fused PET-CT after three months of therapy. Therefore, both patients are still on therapy. This preliminary experience suggests that cetuximab may be a useful therapeutic choice in advanced pre-treated thymic tumors. PMID- 17127336 TI - Role of membrane ion transport proteins in cerebral ischemic damage. AB - Loss of ion homeostasis plays a central role in pathogenesis of ischemic cell damage. Ischemia-induced perturbation of ion homeostasis leads to intracellular accumulation of Ca2+ and Na+ and subsequent activation of proteases, phospholipases, and formation of oxygen and nitrogen free radicals. This signal transduction cascade results in long-term functional and structural changes in membrane and cytoskeletal integrity and eventual cell death. Both ion conductances and ion transporters could affect ion homeostasis. Considerable research effort has been centered on roles of passive fluxes via cation and anion conductances in cerebral ischemic damage. This review will instead focus on the recent studies into the role of secondary active transport proteins in ischemia induced dissipation of ion homeostasis. Secondary active ion transport proteins are a membrane protein-mediated solute transport mechanism that derives its energy from the combined chemical gradients of the transported ions. They are important in maintaining steady-state intracellular ion concentrations. These include Na+-dependent chloride transport (NKCC), Na+/H+ exchange (NHE), and Na+/Ca2+ exchange (NCX). Results from both in vitro and in vivo experimental studies suggest that these ion transport proteins are potential targets to reduce or prevent ischemia-mediated loss of ion homeostasis. PMID- 17127337 TI - Fine urban atmospheric particulate matter modulates inflammatory gene and protein expression in human bronchial epithelial cells. AB - Ambient particulate matter (PM) is known to induce inflammation in the respiratory tract of exposed subjects. The aim of the present study was to detect, in bronchial epithelial cells, candidate inflammatory genes exhibiting transcriptional modifications following urban PM2.5 exposure. Paris urban PM2.5 sampled either at a curbside or a background station in winter and in summer was tested in comparison with diesel exhaust particles (DEP) at 10 microg/cm2 on human bronchial epithelial (16-HBE) cells (18 h of exposure). The gene profiling study performed using a 375 cDNA cytokine expression array highlighted the differential expression of certain genes, three of which were selected as genes of interest: the IL-1 alpha cytokine, the GRO-alpha chemokine, and amphiregulin, a ligand of the EGF receptor. Their increased expression was confirmed by RT-PCR and/or by Northern blotting in bronchial epithelial cells. In the culture medium of particle-treated cultures, increased release of GRO-alpha and amphiregulin was shown. The particle component responsible for protein release varied for the two genes. The organic extract seemed to be mainly involved in amphiregulin expression and secretion, whereas both the aqueous and organic extracts induced GRO-alpha release. In conclusion, in bronchial epithelial cells, Paris PM2.5 increased mRNA and protein expression of GRO-alpha and AR involved in the chemoattraction process and bronchial remodeling, respectively. PMID- 17127338 TI - Improvement in selectivity and storage stability of a choline biosensor fabricated from poly(aniline-co-o-aminophenol). AB - A choline biosensor was fabricated by using electrochemical doping to immobilize choline oxidase in poly(aniline-co-o-aminophenol) film that exhibits a good electric activity over a wide pH range. Using cyclic voltammetry, impedance measurement and scanning electron microscopy characterized the poly(aniline-co-o aminophenol) film doped with choline oxidase. The amperometric detection of choline is based on the oxidation of the H2O2 enzymatically produced on the choline biosensor. The choline biosensor has a lower potential dependence. Thus, its operational potential was controlled at a low potential of 0.40 V (vs.SCE). The response current of the choline biosensor increases with increasing temperature from 277.1 to 308.1 K. An apparent activation energy of 30.8 kJ mol( 1) was obtained. The choline biosensor has a wide linear response range from 1x10(-7) to 1x10(-4) M choline with a correlation coefficient of 0.9999 and has a high sensitivity of 127 microA cm(-2), at 0.40 V and pH 8.0. The response time of the biosensor is 15-25 s, depending on the applied potentials. An apparent Michaelis constant and an optimum pH for the immobilized enzyme are 1.8 mM choline and 8.4, respectively, which are very close to those of choline oxidase in solution. The effect of selected organic compounds on the response of the choline biosensor was studied. Together, these findings show that the choline biosensor exhibits a better selectivity to interfering species and a better storage stability. PMID- 17127339 TI - Activation of the mitochondrial pathway of apoptosis by oxysterols. AB - A significant fraction of cholesterol that accumulates in atherosclerotic lesions is actually oxidized to yield a number of derivatives, named oxysterols, which are provided with much stronger biochemical effects than the parental compound. Of note, an increasing bulk of studies is giving evidence of accumulation of oxysterols in a number of other chronic disease processes including quite common neurodegenerative diseases. In particular, defined cholesterol oxidation products, among those of main interest in pathophysiology, may strongly activate the mitochondrial pathway of apoptotic death. Modulation by oxysterols of various pro- and anti-apoptotic molecules involved in that pathway are hereafter examined under the light of the most recent relevant literature. PMID- 17127340 TI - Chromosomal deletions in bladder cancer: shutting down pathways. AB - Bladder cancer is one of the most common cancers in the world, leading to approximately 145,000 deaths annually. Bladder cancer is typically managed by surgical removal of the tumor; however, the recurrence rate is disappointingly very high, often requiring systemic chemotherapy. Improvement in the diagnosis and prognosis of bladder cancer will only come from a comprehensive understanding of the genetic factors that lead to its development. In this review, we focus on the chromosomal deletions that contribute to the downregulation of tumor suppressor pathways in bladder cancer. Chromosomal deletions are not a random event, since bladder cancer progression has been associated with specific chromosomal deletions and this progression correlates with specific stages of tumor development. The most commonly found chromosomal deletion in all stages of bladder cancer involves deletions in chromosome 9, resulting in the loss of three genes encoding proteins that activate the Rb and p53 tumor suppressors. Additionally, chromosome 9 harbors the TSC1 tumor suppressor which downregulates the well-known anti-apoptotic Akt/mTOR pathway. Hence, deletions on one chromosome may have a crucial influence on the initial steps in tumor development. Other deletions targeting the tumor suppressors Rb, p53, FHIT and LZTS1 occur at later stages of tumor development. Considering the central importance of these tumor suppressor pathways in the formation and evolution of tumors, the time has come to evaluate available drugs in bladder cancer that target the positive regulators of these pathways. PMID- 17127341 TI - Mixed cytokine profile during active cutaneous leishmaniasis and in natural resistance. AB - Most studies on immune response in human cutaneous leishmaniasis evaluate patients with active disease in comparison with healthy uninfected controls or patients that have had the lesions healed, however, little is known about the immune response associated with natural resistance. In this paper we evaluate the cytokine expression patterns of T-cells and the plasmatic levels of nitrite and nitrate in patients with localized cutaneous leishmaniasis (LCL) as well as endemic non-infected individuals with positive (MST) and negative (NI) Montenegro skin test, without previous history of leishmanial lesions. Our results demonstrated an increased number of IFN-gamma+ and TNF-alpha+ T-cells and high level of plasma nitrite and nitrate in LCL patients. Moreover, we have observed that early in infection (LCL equal/less than 60 days of lesion evolution), Leishmania patients present predominance in IL-4+ and IL-10+ T-lymphocytes. However, this is a transitory phenomenon, since patients with older lesions (LCL more than 60 days of lesion evolution) show a predominant Type-1 immune profile, suggesting that disease development may depend on a transient deregulation of T cell response, during the initial phase of infection. Interestingly MST displayed a basal mixed Type-0 cytokines profile. However, the low frequency of IL-4+ T cells, high IFN-gamma+/IL-10+ cell ratio as well as elevated nitrite and nitrate plasma levels observed in MST, suggested that despite basal levels of cytokines, a high proportion of Type-1 over Type-2 cytokines would count to prevent parasite growth and lesion development. PMID- 17127342 TI - The GCK II and III subfamilies of the STE20 group kinases. AB - The Ste20 (sterile 20) proteins are a large family of serine/threonine kinases. Since their discovery a growing body of evidence has implicated them in the regulation of signaling pathways governing cell growth, cell differentiation cell death and cell volume. Approximately 30 human members have been identified based on the high degree of homology of their catalytic domain to that of the Ste20p from Saccharomyces cerevisiae. In addition to the conserved regions, there are also regions of sequence that make each of them unique. In this review we will focus on two subfamilies of the group, GCK-II and GCK-III, families that are closely related but, again, unique in their structural features and biological functions. Herein, we will present what we hope will be the current state of knowledge about these kinases, and discuss what remains to be done in order to better understand their activity and regulation. PMID- 17127343 TI - Hoxb-5 down regulation alters Tenascin-C, FGF10 and Hoxb gene expression patterns in pseudoglandular period fetal mouse lung. AB - Organ-specific patterning is partly determined by Hox gene regulatory interactions with the extracellular matrix (ECM), cell adhesion and fibroblast growth factor (FGFs) signaling pathways but coordination of these mechanisms in lung development is unknown. We have previously shown that Hoxb-5 affects airway patterning during lung morphogenesis. Hoxb-5 regulation in fetal lung affects ECM expression of tenascin-C and alters FGF10 spatial and cellular expression. To test this hypothesis, gestational day 13.5 (Gd13.5) fetal mouse lung fibroblasts and whole lungs were cultured with Hoxb-5-specific small interfering RNA (siRNA). Western blots showed that siRNA-down regulation of Hoxb-5 led to decreased tenascin-C and FGF10 and was associated with increased Hoxb-4 and decreased Hoxb 6 protein levels. Hoxa-5 protein levels were not affected. Hoxb-5 siRNA-treated whole lung cultures had a significant decrease in total lung and peripheral branching region surface area. Immunostaining showed negligible levels of Hoxb-5 protein and tenascin-C, and loss of FGF10 spatial restriction. We conclude that Hoxb-5 helps regulate lung airway development through modulation of ECM expression of tenascin-C. ECM changes induced by Hoxb-5 may affect mesenchymal epithelial cell signaling to alter spatial and cellular restriction of FGF10. Hoxb-5 may also affect lung airway branching indirectly by cross regulation of other Hoxb genes. PMID- 17127344 TI - The transport of glutamine into mammalian cells. AB - Glutamine has many important functions in mammalian cells, and glutamine transport across cell membranes has accordingly been extensively studied. In the past few years a number of important glutamine transport proteins have been sequenced and their molecular properties have been characterised. In general, four major transporters are important physiologically. These are known as (i) SNAT3 (System N) which is important in glutamine uptake in periportal cells in liver and in across the basolateral membrane of renal proximal tubule cells and is also involved in glutamine release by liver perivenous cells and by astrocytes; a variant of this protein catalyses glutamine release from skeletal muscle. (ii) SNAT1 (a specific System A sub-type) which is important in glutamine uptake by neuronal cells (iii) ASCT2 which is essential for glutamine uptake by rapidly growing epithelial cells and tumour cells in culture and (iv) the recently discovered brush border membrane transporter B0 AT1 (SLC6A19). Recent studies considered both the importance of ASCT2 in tumour cell growth and the regulation of ASCT2 expression. In SK-Hep hepatoma cells, knockdown of ASCT2 using antisense mRNA has been shown to cause apoptosis. Expression of the ASCT2 transporter in HepG2 hepatoma cells is stimulated by glutamine by a pathway involving the promoter element AGGTGAATGACTT which binds FXR/RXR dimers. PMID- 17127345 TI - Control of brain glutamine synthesis by NMDA receptors. AB - Glutamine synthetase (GS) is involved in important processes in brain: modulation of the turnover of glutamate through the glutamate-glutamine cycle, detoxification of ammonia and, under certain circumstances, modulation of brain edema. Modulation of GS activity in brain is therefore important and its impairment or saturation may have pathological consequences. In this review we summarize the data showing that GS in brain is modulated by NMDA receptors and nitric oxide. Blocking NMDA receptors or nitric oxide synthase in vivo increases GS activity and glutamine content in brain, indicating that tonic activation of NMDA receptors and nitric oxide synthase maintain a tonic inhibition of GS. NMDA receptor-mediated activation of nitric oxide synthase is responsible only for part of the inhibition of GS. Other sources of nitric oxide also contribute to tonic inhibition. The inhibition is due to a covalent modification of GS, likely nitration of tyrosine residues. This modification would be reversible and it would be an enzyme that denitrosylate or denitrate GS. Moreover, GS would not be working at maximum rate and its activity may be increased pharmacologically by manipulating NMDA receptors or nitric oxide content. This may be useful for example to increase ammonia detoxification in brain in hyperammonemic situations. PMID- 17127346 TI - Transglutaminase 2 in neurodegenerative disorders. AB - Type 2 transglutaminase (TG2) is a calcium-dependent acyltransferase which also undergoes a GTP-binding/GTPase cycle even though it lacks any obvious sequence similarity with canonical GTP-binding (G) proteins. As an enzyme which is responsible for the majority of transglutaminase (TG) activity in the brain, TG2 is likely to play a modulatory role in nervous system development and has regulatory effect on neuronal cell death as well. Most importantly, numerous studies have presented data demonstrating that dysregulation of TG2 may contribute to the pathogenesis of many neurodegenerative disorders, including Huntington's disease, Alzheimer's disease, Parkinson's disease and amyotrophic lateral sclerosis as well as nervous system injuries. Although TG2's involvement in these disease conditions is strongly suggested by various findings, such as the increase of TG2 mRNA expression, protein level and TG activity in the pathological process of these neurodegenerative disorders, as well as the therapeutic effect of TG2 genetic deletion in animal models of Huntington's disease, the precise mechanism underlying TG2's role remain unclear. TG2 was originally proposed to contribute to the pathogenesis of these diseases by facilitating the formation of insoluble protein aggregates, however recent findings clearly indicate that this is likely not the case. Nonetheless, there is data to suggest that TG2 may play a role in neurodegenerative processes by stabilizing toxic oligomers of the disease-relevant proteins, although further studies are needed to validate these initial in vitro findings. PMID- 17127347 TI - MutS homologues hMSH4 and hMSH5: diverse functional implications in humans. AB - The DNA mismatch repair (MMR) pathway is one of the most critical genome surveillance systems for governing faithful transmission of genetic information during DNA replication. The functional necessity of this pathway in humans is partially reflected by the tight link between MMR gene mutations and the development of hereditary nonpolyposis colorectal cancer. Increasing evidence has suggested a broad involvement of MMR proteins in various aspects of DNA metabolism beyond the scope of DNA mismatch correction, such as in the processes of DNA damage response and homologous recombination. Though evidence is presently lacking for potential functional involvement of hMSH4 and hMSH5 in MMR, these two proteins are thought to play roles in meiotic and mitotic DNA double strand break (DSB) repair and DNA damage responses in human cells. PMID- 17127348 TI - Corticotrophin releasing hormone and the timing of birth. AB - Corticotrophin-releasing hormone (CRH) is the hypothalamic peptide that controls the function of the pituitary-adrenal axis in response to stress. CRH is also expressed abundantly in the human placenta and is present in high concentrations in maternal and fetal plasma during late pregnancy. During pregnancy, CRH derived from the placenta is thought to play a crucial role in the regulation of fetal maturation and the timing of delivery, and CRH has also been implicated in the control of fetal-placental blood flow. Elevated CRH concentrations, as compared with gestational age matched controls, occur in patients in preterm labour. The exponential curve depicting the CRH increase is shifted to the left in women who will subsequently deliver preterm and to the right in women who will deliver post dates. This has led to the suggestion that CRH production is linked to a placental clock which determines the length of gestation. Clinically, maternal plasma CRH concentrations may be useful in identifying women at high risk of preterm delivery and CRH antagonists may be useful in preventing preterm labour. As significant CRH production by the placenta is restricted to primates, future research must take into account the species specificity of the mechanisms regulating parturition. A number of significant gaps remain in our knowledge of the function of this peptide in pregnancy. This review examines the current evidence regarding the role of CRH in human parturition. PMID- 17127349 TI - T cell immunotherapy. AB - Adoptive T cell immunotherapy is the isolation of tumor-specific T cells from a cancer patient, in vitro activation and expansion of these T cells, and re infusion of the T cells to the patient. In a small percentage of patients with tumor types susceptible to immune modulation, adoptive therapy has proven to be highly effective. The use of adoptive therapy has several limitations which are being actively investigated today. T cells from various sources are being isolated for adoptive therapy: lymphocytes can be isolated from tumor lesions, from lymph nodes draining the tumor or a tumor vaccine site, or from peripheral blood lymphocytes stimulated with tumor antigens in vitro. Recent advances in T cell therapy include enhanced efficacy of T cell therapy following non myeloablative chemotherapy and genetic modification of T cells for use in adoptive therapy. Clinical trials using gene-modified T cells with improved activation, lifespan, and tumor targeting are on the horizon. It is likely that adoptive immunotherapy will remain a fertile area for investigation resulting in advances in the fields of T cell biology and gene therapy. Adoptive therapy for cancer will become widespread only after its clinical benefit for sizeable patient populations has been established. PMID- 17127350 TI - Marine invertebrate mitochondria and oxidative stress. AB - The marine environment confronts its inhabitants with a wide variety of O2 concentrations, as well as with fluctuations of small scale local O2 availability over time. This review analyzes the respiratory response of marine animal ectotherms to fluctuating environmental O2 availability in their specific habitats and reactive O2 species generation under environmental stress, with a special emphasis on temperature. Specifically we compare mitochondrial functioning and reactive O2 species formation in these animals to what is known from endothermal, mammalian species. Among the strategies employed by marine invertebrates to maintain tissue PO2 low and within tolerable margins, the possible role of mitochondrial oxyconformity to control cellular PO2 in water breathers is discussed. Mitochondrial generation of reactive O2 species in ectotherms has been shown to depend on the magnitude of the mitochondrial membrane potential and the mitochondrial H+ leak. Alternative mitochondrial oxidases are described in marine ectotherms and might add to ameliorate reactive O2 species formation. The effect of nitric oxide, which in mammals controls the reduction state of the electron transport by lowering cytochrome oxidase O2 affinity, remains to be investigated in marine invertebrates. A new concept is proposed, showing how the cross talk of reactive O2 species in metabolically low marine invertebrates could support their outstanding hypoxia tolerance under non stressed conditions. PMID- 17127351 TI - Melatonin role in the mitochondrial function. AB - Melatonin is an ancient molecule present in unicellular organisms at the very early moment of life. Initially identified as a secretory product of the pineal gland in mammals and in other species, it was considered a hormone related to reproduction. The evidence that melatonin is produced in many organs and tissues of the body, reaching concentrations higher than in the blood, support the multiplicity of the melatonin actions. The best-known actions of melatonin, currently supported by experimental and clinical data, include antioxidant and anti-inflammatory abilities, some of them involving genomic regulation of a series of enzymes. Besides, melatonin displays anticonvulsant and antiexcitotoxic properties. Most of the beneficial consequences resulting from melatonin administration may depend on its effects on mitochondrial physiology. The physiological effects of melatonin on normal mitochondria, its role to prevent mitochondrial impairment, energy failure, and apoptosis in oxidatively-damaged mitochondria, and the beneficial effects of the administration of melatonin in experimental and clinical diseases involving mitochondrial dysfunction and cell death, are revised. PMID- 17127352 TI - Nitric oxide and oxygen metabolism in inflammatory conditions: sepsis and exposition to polluted ambients. AB - Nitric oxide and cytokines constitute the molecular markers and the intracellular messengers of inflammatory conditions which are derived from the activation of the NF-kappaB pathway and the transcription of proinflammatory genes. Sepsis occurs with an exacerbated inflammatory response that damages tissue mitochondria and impairs bioenergetic processes. One of the current hypotheses for the molecular mechanisms underlying the complex condition of sepsis is that enhanced NO production by mtNOS leads to excessive peroxynitrite production and protein nitration in the mitochondrial matrix, causing mitochondrial dysfunction and organ failure. The mechanism of particulate matter-health effects are believed to involve inflammation and oxidative stress. Components in particles that elicit inflammation are poorly investigated, although recent research points out to the contribution of compositional elements and particle size. Nitric oxide and reactive oxygen species appears to be involved in the inflammatory conditions associated to particulate matter inhalation. PMID- 17127353 TI - The mechanism of cytochrome C oxidase inhibition by nitric oxide. AB - The basic biochemistry of the inhibition of cytochrome oxidase by NO is reviewed. Three possible mechanisms that include the binding of NO to the fully reduced Fe(a3)-Cu(B) site, to the semi-reduced Fe(a3)-Cu(B) site, and to the fully oxidized Fe(a3)-Cu(B) site are confronted with the experimental data. Mathematical models are used to facilitate the analysis and to solve puzzling observations concerning the NO inhibition of cytochrome oxidase. It is concluded that the inhibition of cytochrome oxidase by NO is mixed, having both competitive and uncompetitive components, but under physiological electron flows the competitive component is largely predominant. The physiological and pathological relevance of this inhibition is briefly discussed. PMID- 17127354 TI - Mitochondrial toxins and neurodegenerative diseases. AB - The selective loss of a particular subset of neurons is a common feature of neurodegenerative disorders. A failure in respiratory chain complex activities in mitochondria seems to be a causative factor. The aim of this review is to describe the most important toxins affecting the mitochondrial function, which could be involved in the incidence of some of these diseases: MPTP, rotenone and 3-nitropropionic (3-NPA). PMID- 17127355 TI - Mitochondrial oxidant generation is involved in determining why females live longer than males. AB - Females live longer than males in many mammalian species, including humans. This natural phenomenon can be explained on the basis of the mitochondrial theory of aging. Mitochondria are a major source of free radicals in cells. Mitochondria from female rats generate half the amount of hydrogen peroxide than those of males and have higher levels of mitochondrial reduced glutathione. The latter is due to females behaving as double transgenic in over-expressing antioxidant enzymes. Estrogens bind to the estrogen receptors and subsequently activate the mitogen activated protein (MAP) kinase and nuclear factor kappa B (NFkappaB) signalling pathways, resulting in an upregulation of antioxidant enzymes. Moreover, the 16S rRNA expression, which decreases significantly with aging, is four times higher in mitochondria from females than in those from males of the same chronological age. On the contrary, the oxidative damage of mitochondrial DNA is fourfold higher in males than in females. Ovariectomy abolishes the gender differences between males and females and estrogen replacement rescues the effect of ovariectomy. The challenge for the future is to find molecules that have the beneficial effects of estradiol, but without its feminizing effects. Phytoestrogens or phytoestrogen-related molecules may be good candidates to meet this challenge. PMID- 17127356 TI - Oxygen delivery to the tissues and mitochondrial respiration. AB - In mammals, the O2 transport from the inspired air to the tissues is made by convective and diffusive mechanisms. The convective mechanisms are provided by the cardio-respiratory system and comprised by the basic variables of cardiac output and blood O2 content. Microcirculation in arterioles and capillaries is adjusted to match the O2 demand of local tissues. Endothelium-generated NO diffuses to the smooth muscle of microvessels and produces vasodilation that increases circulatory time in the capillaries and allows a more effective O2 extraction in the tissues. Once within the tissue, O2 diffuses to mitochondria where it is reduced in an exergonic process coupled to ATP synthesis. Both, O2 and ATP are the two most homeostatic intracellular species. In heart and muscle, both species show unchanged levels with 25-100 times increases in work load and ATP turnover rate. The linear rates of O2 uptake shown by tissue slices and perfused organs are interpreted as a fast switching of mitochondria between metabolic state 3 (with a fast rate of O2 uptake and ATP synthesis) and state 4 (with a slow rate of O2 uptake and no ADP phosphorylation). Endogenous mitochondrial NO, produced by mtNOS, sustains the concept of a physiological functional activity of this enzyme in regulating mitochondrial and cellular O2 uptake. PMID- 17127357 TI - Nitric oxide and mitochondria. AB - Nitric oxide (NO) and its derivatives (reactive nitrogen species) have multiple effects on mitochondria that impact on cell physiology and cell death. Mitochondria may produce and consume NO and NO stimulates mitochondrial biogenesis, apparently via cGMP upregulation of transcriptional factors. NO inhibits mitochondrial respiration via: (A) an acute and reversible inhibition of cytochrome oxidase by NO in competition with O2, and (B) irreversible inhibition of multiple sites by reactive nitrogen species. NO is a potent vasodilator (via cGMP), increasing O2 and respiratory substrate supply to mitochondria. NO stimulates reactive oxygen and nitrogen species production from mitochondria via respiratory inhibition, reaction with ubiquinol and reaction with O2 in the membrane. NO can induce apoptosis, mainly via oxidative stress. NO induces necrosis, mainly via energy depletion. Reactive nitrogen species activation of the mitochondrial permeability transition pore may cause apoptosis or necrosis. NO may protect against mitochondria-mediated cell death by multiple mechanisms. PMID- 17127358 TI - Brain nitric oxide synthases and mitochondrial function. AB - Nitric oxide is a small signaling molecule, which may act as a neurotransmitter and neuromodulator, exerting a regulatory effect on neuronal function. It can diffuse from its site of synthesis to different intra and extracellular compartments, being therefore present in the pre-synaptic, synaptic and post synaptic spaces. Recently, a NOS located in the mitochondria (mtNOS) has been observed in different brain regions, responsible for the production of NO in these organelles and identified as nNOS. A regulatory effect of NO on mitochondrial function was described in brain mitochondria, where NO acts mainly by inhibiting cytochrome oxidase activity. Hippocampal mitochondrial dysfunction and decreased mtNOS activity and expression were reported in association with ultrastructural damage in an experimental model of hepatic encephalopathy. Enriched environment exposure preserved the aged animals from spatial cognition impairment; also environment and training modulated neuronal plasticity in pre pubertal rats through NO-dependent mechanisms. In addition, brain cortical mitochondrial respiration and mtNOS activity and expression were analyzed as function of age. Mitochondrial NO production showed a decreasing tendency as a function of age. These results are in accordance with the protein expression analyzed by Western Blot of mitochondrial fractions which was 6.5 times higher in 1 month aged rats as compared with 14 old animals. Concomitant with these results, a clear increasing oxygen uptake tendency in state 3 respiration was observed, meanwhile only a slight increase was observed in state 4. All these results seems to be clearly related with the reversible and concentration dependent attenuation of the respiratory chain by NO. PMID- 17127359 TI - The biological significance of mtNOS modulation. AB - In the last years, nitric oxide synthases (NOS) have been localized in mitochondria. At this site, NO yield directly regulates the activity of cytochrome oxidase, O(2) uptake and the production of reactive oxygen species. Recent studies showed that translocated neuronal nitric oxide synthase (nNOS) is posttranslationally modified including phosphorylation at Ser 1412 (in mice) and myristoylation in an internal residue. Different studies confirm that modified nNOS alpha is the main modulable isoform in mitochondria. Modulation of mtNOS was observed in different situations, like adaptation to reduced O(2) availability and hypoxia, adaptation to low environmental temperature, and processes linked to life and death by effects on kinases and transcription factors. We present here evidence about the role of mtNOS in the analyzed conditions. PMID- 17127360 TI - The interaction between the renin-angiotensin system and peroxisome proliferator activated receptors: a hypothesis including the participation of mitochondria in aging. AB - The objective of improving health is intimately associated with preventing and delaying age-related diseases. Nutritional and pharmacological approaches aimed at retarding aging are uncovering mechanisms, whose definitive roles in cell and tissue physiology need to be defined. In this article we hypothesize that peroxisome proliferator activated receptor (PPAR)-modulation is a pivotal process that underlies the association between mitochondria and the renin-angiotensin system (RAS) in aging. This hypothesis is based on several lines of evidence suggesting that: a) mitochondrial function and oxidant production are active participants in the aging process; b) PPARs, by regulating mitochondrial function and uncoupling proteins (UCP), seem to play a major role in the age-retarding effects of caloric restriction; c) RAS inhibition delays the deleterious effects of aging and also upregulates PPARs; and d) a number of physiological and molecular events that occur in experimental caloric restriction, and experimental and clinical RAS inhibition, involve changes in mitochondrial functions. PMID- 17127361 TI - Mitochondrial ferritin in animals and plants. AB - Ferritins play a role in preventing Fe toxicity because of their ability to sequester several thousand Fe atoms in their central cavity in a soluble, non toxic bioavailable form. The identification of ferritin in mitochondria, an organelle with a constant generation of O2(-) as a by-product of the electron transfer, and the presence of a mitochondrial nitric oxide synthase activity opened up brand new metabolic interactions to be analyzed. In spite of cytosolic ferritins in mammals being ubiquitous, mitochondrial ferritin (mtF) expression is restricted to the testis, neuronal cells, islets of Langerhans, and as recently described to mice normal retinas. None was detected in major storage organs such as liver and spleen. MtF has about 80% identity to cytosolic H-chain and 55% to L chain in its coding region. There has been reported some differences in the Fe binding and oxidation properties between mtF and cytosolic H-ferritin suggesting that mtF functions differently as an Fe storage protein within the mitochondria and perhaps has other function(s) in Fe homeostasis as well. Recently it was also presented evidence for the presence of ferritins in plant mitochondria. The understanding of the role of mitochondrial ferritin in Fe oxidative metabolism may be useful in approaching clinical situations such as the treatment of Friedreich's ataxia, X-linked sideroblastic anemia, and in other neurodegenerative disorders. PMID- 17127362 TI - Mitochondrial nitric oxide synthase. AB - Nitric oxide (NO) regulates several cellular functions via reversible regulation of mitochondrial respiration. Nitric oxide also reacts with mitochondrial superoxide anion to produce the potent oxidative species peroxynitrite that irreversibly hinders mitochondrial activities. Recent findings demonstrating that mitochondria produce NO via mitochondrial NO synthase (mtNOS) has intrigued several laboratories revealing crucial roles for mtNOS-derived NO and peroxynitrite in regulating the functions of mitochondria, cells and organs. The present article reviews the current understanding of the interactions between mitochondria, and NO and peroxynitrite. PMID- 17127363 TI - Pesticides and impairment of mitochondrial function in relation with the parkinsonian syndrome. AB - The Parkinsonian syndrome induced by pesticides is associated with the impairment of mitochondrial function. Toxicants that inhibit selectively NADH-dehydrogenase activity, as rotenone or pyridaben, also show a selective inhibition of O2 uptake and respiratory control in rat brain mitochondria in the presence of NAD dependent substrates. The IC50 of rotenone and pyridaben for complex I inhibition were in the range 1.7-2.2 microM. The determination of NADH-cytochrome c reductase, succinate-cytochrome c reductase and cytochrome oxidase activities in rat brain submitochondrial showed again the selective inhibition of Complex I by rotenone and pyridaben, whereas paraquat produced a non-selective inhibition affecting all the respiratory chain complexes. In rat brain mitochondria, rotenone and pyridaben markedly decreased mtNOS functional activity with NAD dependent substrates but not when the substrate was succinate. This observation suggest than mtNOS activity is regulated by the activity of complex I. This regulation and the role of mitochondrial NO diffusion as a signal for mitochondrial biogenesis could have a role in the etiopathology of Parkinson's disease. PMID- 17127364 TI - Coordination of physiologic and toxic pathways in hippocampus by nitric oxide and mitochondria. AB - The hippocampus, one of the most vulnerable regions in the brain, has been implicated in learning and memory formation. However, impairment of hippocampal function is observed during normal aging and neurodegenerative processes. Current evidence suggests that mitochondria and NO participate prominently in cellular signaling in the hippocampus integrating physiologic and toxic pathways. Although all isoforms of nitric oxide synthase are expressed in hippocampal cells, the production of NO in the dependency of glutamate receptors is primarily involved in hippocampal physiopathology. Taking into consideration that the biological impact of NO remains largely qualitative, this review discusses generally the regulation of glutamate-dependent NO production in hippocampus with implications in synaptic plasticity and explores mechanisms by which NO and mitochondria coordinate physiologic and toxic pathways, in particular the excitotoxic NO mitochondrial connection, the excitotoxic-dependent DNA damage and the mitochondrial biogenesis and trafficking. PMID- 17127365 TI - Mitochondrial dysfunction, free radical generation and cellular stress response in neurodegenerative disorders. AB - Protein conformational diseases, such as Alzheimer's, Parkinson's and Huntington's, affect a large portion of aging population. The pathogenic dysfunctional aggregation of proteins in non-native conformations is associated with metabolic derangements and excessive production of reactive oxygen species. Reduction of cellular expression and activity of antioxidant proteins result in increased oxidative stress. Free-radicals derived from mitochondrial dysfunction and from the cyclooxygenase enzyme activity play a role in oxidative damage of brain. Cyclooxygenase also mediates in neuro-inflammation by the production of pro-inflammatory prostaglandins which contribute to brain injury. The pathogenic role of cyclooxygenase has been demonstrated in Alzheimer and Parkinson diseases. The brain responses to detect and control diverse forms of stress are accomplished by a complex network of "longevity assurance processes" integrated to the expression of genes termed vitagenes. Heat shock proteins are a highly conserved system responsible for the preservation and repair of correct protein conformation. Heme oxygenase-1, a inducible and redox-regulated enzyme, is currently considered as having an important role in cellular antioxidant defense. A neuroprotective effect, due to its heme degrading activity, and tissue-specific pro-oxidant effects, due to its products CO and free iron, are under debate. There is a current interest in dietary compounds that can inhibit, retard or reverse the multi-stage pathophysiology of Alzheimer disease, with a chronic inflammatory response, brain injury and beta-amyloid associated pathology. Curcumin and ferulic acid, two powerful antioxidants, the first from the curry spice turmeric and the second a major constituent of fruit and vegetables, have emerged as strong inducers of the heat shock response. Food supplementation with curcumin and ferulic acid is considered a nutritional approach to reduce oxidative damage and amyloid pathology in Alzheimer disease. PMID- 17127366 TI - Mitochondrial oxidative and structural damage in ischemia-reperfusion in human myocardium. Current knowledge and future directions. AB - The sequence of events in heart ischemia-reperfusion has been clearly documented in experimental animal models but not in cardiac surgery patients. The evidence in human studies had not been gathered in a systematic and comprehensive fashion, so as to provide an encompassing picture of the phenomenon. This limits our ability to devise appropriate strategies for optimal perioperative myocardial protection. We present here a review or our experience in myocardial ischemia reperfusion in a historical perspective. From our previous studies we conclude that, although several issues still remain unsolved, there is no doubt that oxygen-free radicals are important contributors to myocardial injury during the reperfusion period of coronary artery bypass surgery. Yet, in spite of this wealth of information, both clinical and experimental, subsequent clinical trials conducted over the last several years with a variety of antioxidant strategies have been largely disappointing. Therefore, the whole paradigm of oxidative stress in cardiac injury needs to be re-evaluated. In this regard, differences between past and current knowledge are discussed, and future directions are traced. We concluded that patients subjected to elective bypass surgery undergo oxidative stress upon reperfusion after cardioplegic arrest; the magnitude of the phenomenon, however, is at present small and may not justify widespread antioxidant therapy. PMID- 17127367 TI - Mitochondrial dysfunction in human pathologies. AB - The integrity of mitochondrial function is fundamental to cell life. The cell demands for mitochondria and their complex integration into cell biology, extends far beyond the provision of ATP. It follows that disturbances of mitochondrial function lead to disruption of cell function, expressed as disease or even death. Mitochondria are major producers of free radical species and also possibly of nitric oxide, and are, at the same time, major targets for oxidative damage. In this review we consider recent developments in our knowledge of how the mitochondrial production of reactive oxygen species (ROS) plays a critical role in several major human pathologies. We will also consider recent advances in our understanding of the molecular mechanisms involved in mitochondrial ROS detoxification. PMID- 17127368 TI - Brain mitochondrial dysfunction in aging: conditions that improve survival, neurological performance and mitochondrial function. AB - Mice with (a) high spontaneous neurological activity, or subjected to (b) moderate exercise or (c) dietary supplemented with high doses of vitamin E from 28 weeks of age to senescence (76 wk of age), showed an increased survival and a retardation in the development of the neurological deficits associated to aging. During aging there was an increase in dysfunctional brain mitochondria, characterized by an increased content of oxidation products and by a diminished functional activity. The mitochondrial oxidative damage observed in adult (52 wk) and senescent mice (76 wk) was partially ameliorated in the groups of animals subject to the mentioned experimental conditions, and this decrease in mitochondrial oxidative damage was related to the improvement in neurological performance. In brain mitochondria, the activities of enzymes that are critical for mitochondrial function (mtNOS, NADH-dehydrogenase, and cytochrome oxidase) decreased progressively during aging and constituted aging markers. Usual clinical recommendations for aged humans, such as increased neurological activity, moderate exercise, and vitamin E supplementation, proved to be effective in increasing mice survival and neurological performances, along with a better mitochondrial function and a lower content of oxidation products. PMID- 17127369 TI - Role of nitric oxide on mitochondrial biogenesis during the ovarian cycle. AB - The mitochondrial changes in the ovary during the ovarian cycle are adapted to a cyclically increased-decreased energy demand. In the proliferative phase, the increased energy needs are sustained by the recruitment of mitochondria in active state 3, by an increased tissue O2 consumption and ATP production and by an increase in the number of mitochondria. In the phase of decreased energy needs, mitochondria-dependent apoptosis reduces tissue and mitochondrial O2 uptake. The ovary morphological changes during the cycle describe a process in which the follicles undergo a clear cycle with two sequential phases: proliferation and apoptosis. The follicular growth stimulated by FSH characterizes a tissue that shows a quick cell proliferation. During the ovarian cycle, tissue and mitochondrial O2 uptakes, mitochondrial mass, mitochondrial NO production and cytochrome oxidase activity exhibit a biphasic pattern, with marked increases in the ovary proliferative phases. Relatively low levels of NO seem to drive the cell signaling for follicle proliferation, whereas relatively high NO levels trigger mitochondria-dependent follicle apoptosis. PMID- 17127370 TI - Oxidative damage lipid peroxidation in the kidney of choline-deficient rats. AB - Phosphatidylcholine is the most abundant phospholipid constituent of cell membranes and choline is a quaternary amine required for phosphatidylcholine synthesis. The impairment of membrane functions is considered as an indication of oxidative damage. In order to kinetically analyze the time course of the pathogenesis of renal necrosis following to choline deficiency in weanling rats, we determined markers of membrane lipid peroxidation (thiobarbituric acid reactive substances; TBARS and hydroperoxide-induced chemiluminescence (BOOH-CL) ) and studied the histopathological damage. Plasma TBARS (t(1/2) = 2.5 days) was an early indicator of systemic oxidative stress, likely involving liver and kidney. The levels of TBARS an BOOH-CL increased by 80% and by 183%, respectively, in kidney homogenates with t(1/2) = 1.5 days and 4 days, respectively. The levels of BOOH-CL were statistically higher in rats fed a choline-deficient diet at day 6, in a mixture of membranes (from plasmatic, smooth and rough endoplasmic reticulum and Golgi), in mitochondrial membranes and in lysosomal membranes. The results indicate that choline deficiency produces oxidative damage in kidney subcellular membranes. Necrosis involved mainly convoluted tubules and appeared with a t(1/2) = 5.5 days. An increase in the production of reactive oxygen species, triggered by NADH overproduction in the mitochondrial dysfunction associated with choline deficiency appears as one of the pathogenic mechanism of mitochondrial and cellular oxidative damage in choline-deficiency. PMID- 17127371 TI - Induction of mitochondrial xanthine oxidase activity during apoptosis in the rat mammary gland. AB - Oxidative stress is an important signal for apoptosis to start. So far the mitochondrial respiratory chain has been considered as the major, if not the only, cause of such stress. Here we report that this is not the case. Xanthine oxidase, a O2(-) and H2O2 generating enzyme which is important in causing significant oxidative stress in the cytosol, is also present in the mitochondrial fraction of rat mammary gland. After weaning, during the involution of the mammary gland, massive apoptosis occurs. Mitochondrial xanthine oxidase activity increases and high mitochondrial H2O2 production takes place. Inhibition of xanthine oxidase activity by allopurinol, a specific inhibitor of xanthine oxidase activity, slows down the involution of the mammary gland due to the decrease in the number of apoptotic cells and prevents the production of H2O2 that occurs during apoptosis. Thus, mitochondrial xanthine oxidase by means of its production of O2(-) and H2O2 can maintain the apoptotic machinery during the involution of the mammary gland after weaning and could be considered necessary to maintain the apoptotic cascade during the physiological involution of tissues. Oxidative stress generated during apoptosis by mitochondria is not only due to the respiratory chain. PMID- 17127372 TI - Mitochondrial dysfunction in human colorectal cancer progression. AB - The classic association between cancer and mitochondrial dysfunction is actually considered as a role of mitochondria in cellular signalling. It is understood that mitochondria, mitochondrial oxidative damage and NO and H2O2 diffusion are involved in the progression of human colorectal cancer. Mitochondria from human colorectal tumors and adjacent non-tumor colon tissues showed a markedly increased oxidative damage with increased contents of TBARS and protein carbonyls. Mitochondrial protein carbonyls was the most sensitive indicator. Oxidative stress and damage was also observed in adjacent non-tumor cells. Mitochondrial activities, as NADH-cytochrome c reductase and cytochrome oxidase, were observed decreased in tumor and in adjacent non-tumor tissue. Cu,Zn-SOD activity decreased by 42% in tumor tissue in the advanced stage as compared with the initial stage, whereas Mn-SOD activity did not change in tumor progression. An increased mtNOS activity (46%) was observed in tumor and non-tumor tissues in the advanced stage of cancer progression. A direct linear relationship between mtNOS and oxidative damage in tumor and non-tumor tissues supports the concept that mitochondrial NO and H2O2 diffuse from tumor to adjacent non tumor tissue signaling for cell death as the classic toxohormones. PMID- 17127373 TI - Mitochondrial function in liver disease. AB - Oxidative stress is involved in the pathogenesis and progression of different liver diseases, such as alcoholic liver disease and biliary cirrhosis. The increased mitochondrial production of O2(-) at complexes I and III, and consequently of H2O2 and other reactive oxygen species (ROS), triggered by NADH overproduction seems the major cause of mitochondrial and cellular oxidative stress and damage in chronic alcoholism. The mitochondrial oxidative stress renders hepatocytes susceptible to ethanol- or acetaldehyde-induced mitochondrial membrane permeability transition (MMPT) and apoptosis. Nitrosative stress contributes to cell death by peroxynitrite formation. The expression of the death receptor ligand CD95 is also up-regulated by acetaldehyde metabolism. Consequently, a dual mechanism, NADH-driven MMPT and CD95-mediated apoptosis, involving in both cases acetaldehyde metabolism and ROS production, operates in ethanol-induced apoptosis. In the biliary cirrhosis induced by chronic cholestasis, liver mitochondria show increased H2O2 production and GSH depletion and oxidation. Dysfunctional hepatocytes, with a loss in mitochondrial cardiolipin and decreased mitochondrial membrane potential evolve during cholestasis to apoptosis. Ursodeoxycholic acid prevents enlargement of this population as well as mitochondrial oxidative stress. Mitochondrial oxidative stress precedes the initiation and execution of hepatocyte apoptosis in chronic alcoholism and biliary cirrhosis. We suggest that overproduction of mitochondrial NADH is the primary cause for the development of alcoholic and non-alcoholic liver disease by a situation of chronic mitochondrial oxidative stress, which should be considered the second hit that renders hepatocytes susceptible to cell injury and apoptosis. PMID- 17127374 TI - Mitochondrial nitric oxide synthase, a voltage-dependent enzyme, is responsible for nitric oxide diffusion to cytosol. AB - The mitochondrial metabolic state regulates the rate of mitochondrial NO production and release to the cytosol. Nitric oxide release of rat heart mitochondria decreased markedly from 2.2 to 1.2 nmol NO/min. mg protein in the state 4 to state 3 transition. The activity of mtNOS, responsible for NO release, is driven by the membrane potential and not by intramitochondrial pH changes. The release of NO by rat liver mitochondria showed an exponential dependence on membrane potential. A similar behavior was reported for heart mitochondrial H2O2 production. The fraction of heart cytosolic NO provided by diffusion from mitochondria is 90%. The intramitochondrial concentrations of L-arginine and NADPH are higher than their KM values, and the changes in their concentrations in the state 4-state 3 transition are not enough to explain the changes in NO release. These data indicate that the redox state of the respiratory chain components regulates H2O2 production and mitochondrial membrane potential modulates NO release, and support the speculation that NO and H2O2 are a biological signal that reports a high mitochondrial energy charge to the cytosol. The marked regulation of mtNOS activity, as a voltage-dependent enzyme and at the physiological range of membrane potentials, makes mtNOS a highly sensitive enzyme that in turn regulates mitochondrial O2 uptake and H2O2 production. PMID- 17127375 TI - Thyroid hormone calorigenesis and mitochondrial redox signaling: upregulation of gene expression. AB - Thyroid hormone (TH, T3) is required for the normal function of most tissues, with major effects on O2 consumption and metabolic rate. These are due to transcriptional activation of respiratory genes through the interaction of T3 liganded TH receptors with TH response elements or the activation of intermediate factors, with the consequent higher rates of mitochondrial oxidative phosphorylation and reactive O2 species (ROS) generation and antioxidant depletion. The genomic effects of TH are accompanied by redox upregulation of the liver expression of cytokines (tumor necrosis factor-alpha [TNF-alpha]), enzymes (manganese superoxide dismutase), and anti-apoptotic proteins (Bcl-2), via a cascade initiated by TNF-alpha produced by Kupffer cells and involving inhibitor of kappa-B phosphorylation and nuclear factor-kappa-B activation. Thus, TH calorigenesis triggers non-genomic effects leading to an expression pattern that may represent an adaptive mechanism to re-establish redox homeostasis and promote cell survival under conditions of ROS toxicity secondary to TH-induced oxidative stress. Mechanisms of expression of respiratory and redox-sensitive genes may be functionally integrated, which could be of importance to understand the complexities of TH action and the outcome of thyroid gland dysfunction. PMID- 17127376 TI - Reactive oxygen species, antioxidants and fish mitochondria. AB - In fishes, irrespective of their thermoregulatory capacity or metabolic rate, the main physiological source of reactive oxygen species (ROS) is mitochondria. During active swimming, ROS is by an large provided by red muscle mitochondria. Other tissues such as lens, liver, heart, swimbladder, roe and blood also afford important ROS production and antioxidant levels in resting fish. A close relationship between structure and function is evident in fish mitochondrion with a surface-to-volume optimization by the size of cristae to maximize electron transfer. The mechanism of fish mitochondrial superoxide anion (O2*-) and ROS production as well as the mechanism of mitochondrial coupling and proton leak seems similar to that of mammals. Contrary to mammalian red cells, fish erythrocytes possess nuclei and mitochondria. The presence of cardiolipin and the absence of cholesterol in fish mitochondrial membranes confer a high structural flexibility. The difference in phospholipid unsaturation may explain the greater proton leak in endotherms compared to thermoconformers. The present review summarizes our current understanding in respect to comparative aspects of fish mitochondrial function, with an emphasis on the adaptations to changes in temperature, O2 availability and O2 consumption, which are generally coupled to changes in antioxidant status and ROS production. Nevertheless, most work on this fascinating area has yet to be done. The literature on the effect of xenobiotics, aquatic contamination, and aquaculture issues are not reviewed. Data on the production of NO and reactive nitrogen species (RNS), on O2 sensing and on the role of ROS and RNS in cell signalling involving fish mitochondria are almost completely lacking in the literature. PMID- 17127377 TI - Impairment of mitochondrial function by particulate matter (PM) and their toxic components: implications for PM-induced cardiovascular and lung disease. AB - Increasing evidence suggests that reactive oxygen species (ROS) and oxidative stress are involved in PM-mediated lung and cardiovascular injury. The physical characteristics and the chemical composition of particulate matter (PM) play a key role in ROS generation in vitro and in vivo. The mitochondria are major subcellular targets for PM as well as a source of ROS production. ROS production is due to interference in mitochondrial electron transfer and PT pore opening by pro-oxidative PM components. Another possible mechanism is direct physical targeting by ambient ultrafine particles that lodge in and destroy mitochondrial structure. An understanding of the mitochondrial effects of PM is key in understanding the mechanisms of PM-induced adverse health effects. PMID- 17127378 TI - Survivin is not only a death encounter but also a survival protein for invading tumor cells. AB - Cell proliferation and cell death pathways meet at a pivotal crossroad, crucial to maintain normal homeostasis and to eliminate dangerous cells before they start dividing. Survivin (SVV) is an intriguing and fascinating protein at this crossroad that interfaces life and death, through its dual role in facilitating cell division and encountering apoptosis. SVV's prominent expression in essentially all human malignancies, and low or no expression in most normal tissues, suggests that it would be an ideal target for cancer-directed therapy. However, SVV has been recently described as a target for fine tuning by alternative splicing mechanism generating five defined splice variants and a number of other uncharacterized/bizarre isoforms. This diversity indicates that SVV, in addition to its known functions in tumorgenesis, angiogenesis and cardiovascular diseases, might be associated with other unknown functions. Intriguingly, new accumulating evidence from our own work and others, suggest a novel role for SVV in the mechanisms of tumor invasion and metastasis. The SVV pathway has now provided tangible opportunities for targeted, rational cancer therapy. It is therefore an attractive and promising therapeutic target not only for cancer but also for other diseases. Although a number of studies utilizing SVV as an anti-cancer strategy are well underway, further investigation into the exact molecular interactions underpinning its functions is critical for the success of such trials. Impeding development of safe and effective SVV antagonists for clinical use is due to a lack of understanding the molecular mechanisms by which SVV differentially affects apoptosis and cell division in both normal and malignant cells. In this report, in addition to reviewing the SVV known functions, we discuss the newly proposed mechanisms by which SVV might serve as a survival tool for invading tumor cells. PMID- 17127379 TI - HPA axis activity and neuropathogenesis in HIV-1 clade C infection. AB - A disturbance in the activity of the hypothalamic-pituitary-adrenal (HPA) axis has been reported among individuals with HIV-1 infection. However, these studies have been carried out in the West where the infecting clade is clade B. HIV-1 infection is rapidly spreading in various parts of South East Asia, including India, where the HIV-1 infecting clade is largely clade C. An investigation of HPA axis activity in this type of infection is warranted since there are many structural differences between clades B and C. This study was carried out to investigate whether HIV-1 infection clade C interferes with the functions of the hippocampus and thereby affects the HPA axis. We tested the hypothesis that when hippocampus activity is disturbed, it leads to the development of neuropathogenesis in HIV-1 C-clade infected individuals. This study included asymptomatic HIV-1 seropositive individuals (n=117) and, age-matched, HIV-1 seronegative controls (n=29). Neuroendocrine function of the HPA axis was evaluated using plasma levels of cortisol, ACTH, and DHEA-S, both in the morning (0800-1000 hr) and evening (2000-2200 hr). A significant elevation of cortisol levels during A.M. and P.M. hours was observed in HIV-1 infected individuals when compared to the controls. Interestingly, no significant change in ACTH level was observed in HIV-1 seropositive subjects, either during A.M or P.M. Elevated levels of cortisol in HIV-1 seropositive subjects appear to be independent of ACTH and may be the result of a defective negative feedback mechanism. On the other hand, a significant decrease in the plasma levels of DHEA-S was observed during A.M. and P.M. hours in HIV-1 infected individuals, leading to an increased cortisol to DHEA-S ratio. Since increased levels of cortisol and decreased levels of DHEA-S are related to the development of neuropathogenesis, it is hypothesized that a study of the development of neurocognitive deficits among HIV-1 seropositive individuals in India is warranted. PMID- 17127380 TI - The importance of vaccination. AB - We have vaccines for nearly thirty of the more than seventy infectious diseases which are pathogenic for humans. Most of the vaccines, especially those to prevent childhood diseases, are highly effective with a high safety profile. Vaccines are being developed against many of the other bacteria and viruses, and some parasites. Occasionally, a new vaccine has to be withdrawn because of unexpected side effects. Smallpox remains the only infectious disease to have been eradicated. The Global Program to eradicate poliomyelitis initiated in 1988, has unfortunately run into difficulties. A few children immunised with the Sabin oral vaccine fail to clear the virus which can mutate over some years into a pathogenic form and spread rapidly unless large vaccination programs are re introduced. Of major concern are emerging and re-emerging infectious diseases, especially HIV, for which there is currently no vaccine. Fortunately, new techniques are becoming available making it possible to consider developing vaccines based on inducing strong cell-mediated immune responses to control the agent's replication when antigenic variation in surface antigens (e.g. HIV, influenza) makes classical techniques based on induction of antibody responses less attractive. PMID- 17127381 TI - Buprenorphine in long-term control of chronic pain in cancer patients. AB - The aim of this randomized open-label prospective study was to evaluate the analgesic activity of buprenorphine in a transdermal formulation for cancer chronic pain control versus sustained-release morphine, in all cases combined with oral tramadol. A transdermal system with 35 microg/h buprenorphine was applied to the first group of patients (BT); the second group received 60 mg/day of sustained-release morphine (MT). In both groups oral tramadol was administered to a maximum of 200 mg daily, in case of need. The administration of transdermal buprenorphine versus morphine resulted in significant differences in the physical pain (P = 0.01), mental health (P = 0.03) and vitality (P = 0.001). These data indicated that the BT group showed an improvement of pain and a positive effect on the quality life. PMID- 17127382 TI - Lactobacillus delbrueckii as a potential skin adjuvant for induction of type 1 immune responses. AB - We evaluated the ability of Lactobacillus delbrueckii UFV H2b20, a probiotic candidate, to stimulate the production of inflammatory cytokines and to induce macrophage activation and Th1 differentiation in peripheral blood mononuclear cells (PBMC) from healthy volunteers. Our results show that PBMC stimulated with heat-killed Lact. delbrueckii produced elevated levels of IL-12, IFN-gamma and TNF-alpha but no IL-10. IFN-gamma production was IL-12 dependent with NK cells as the main source. Furthermore, PBMC infected with Leishmania amazonensis presented elevated microbicidal activity when co-incubated with Lact. delbrueckii. Finally, Lact. delbrueckii was capable of inducing in vitro differentiation of L. amazonensis-specific Th1 cells. These findings suggest that this probiotic may be used as an adjuvant in vaccination protocols. PMID- 17127383 TI - The interference of rosmarinic acid in the DNA fragmentation induced by osmotic shock. AB - The induction of cell death in human erythroleukemic cells (K562) by sorbitol shows the typical apoptotic changes in ultrastructural morphology, including blebbing, chromatin condensation and nuclear membrane breakdown. Using a cytofluorimetric approach, we found that sorbitol induced production of reactive oxygen species (ROS) followed by DNA fragmentation in leukemic cells. In this study, we investigated effect of curcumin and rosmarinic acid on cell viability in three different cell lines: erythroleukemia K562, papillary NPA, and anaplastic ARO thyroid cancers. Curcumin was able to induce apoptosis in a concentration- and time dependent manner in three cell lines, while rosmarinic acid was less effective on this process. To examine this possibility in cellular system, this study evaluated the capacities of both compounds acting as antioxidant inhibiting sorbitol-induced apoptosis. K562, NPA and ARO cells were pre-incubated with 25 microM rosmarinic acid to allow the uptake and then the cell lines were treated with 1 M sorbitol. Afterwards, the cells were subjected to agarose gel electrophoresis to assess the DNA fragmentation. In conclusion, the antioxidant activity of rosmarinic acid is able to inhibit sorbitol-induced apoptosis. PMID- 17127384 TI - Cerebral tissue oxygenation and oxidative brain injury during ischemia and reperfusion. AB - The brain requires glucose and oxygen to maintain neuronal metabolism and function. Cerebral ischemia causes heterogeneous changes in tissue oxygenation and cellular metabolism, with a region of decreased blood flow, the penumbra, surrounding a severely damaged ischemic core. Because oxygenation is central in ischemic neuronal death, it is critical to understand exactly what actual changes occur in interstitial oxygen tension (pO2) in ischemic regions during stroke, particularly the penumbra and ischemic core. Cerebral ischemia induces a complex series of molecular pathways involving signaling mechanisms, gene transcription, and protein formation. Free radicals and oxidative stress have been suggested to be involved in each of the steps in the injury cascade. The goal of this review paper is to summarize the current literature concerning our understanding about cerebral tissue oxygenation changes after cerebral ischemia and reperfusion, the subsequent cellular and physiological changes in response to alteration in tissue oxygenation, and treatment strategies utilized to minimize the detrimental effects caused by stroke. PMID- 17127385 TI - Role of the prostaglandins in labour and prostaglandin receptor inhibitors in the prevention of preterm labour. AB - Parturition is composed of five separate but integrated physiological events: fetal membrane rupture, cervical dilatation, myometrial contractility, placental separation, and uterine involution. Prostaglandins (PGs) have central roles in each of these events, but the most studied is myometrial contraction. Elevated uterine PGs or the enhanced sensitivity of the myometrium to PGs leads to contractions and labour. The primary regulator of PG synthesis is the mRNA expression of PG H Synthase (PGHS-2 or COX-2). Given the central role of PGs in labour, this enzyme becomes an obvious therapeutic target for the prevention of preterm labour, the major cause of perinatal mortality and morbidity. Unfortunately, even though the non-steroidal anti-inflammatory drugs (NSAIDs), which inhibit PGHS, are usually successful in suppressing preterm labour or prolonging pregnancy in animal and human studies, the NSAIDS have had adverse effects on fetal physiology and development. Therefore, other means to suppress PG synthesis or action to arrest preterm labour need to be investigated. The PGF2alpha receptor, FP, may prove to be a reasonable target for tocolysis. FP mRNA increases in the mouse uterus at preterm birth, whereas PGF2alpha concentrations do not increase, suggesting elevated uterine sensitivity to contractile agonists is one mechanism for preterm labour initiation. New data shows that administration of a specific FP antagonist, Theratechnologies (THG) 113.31, delays preterm birth in mice and sheep with no observable maternal or fetal side effects. Hence antagonizing PG action offers new hope for delaying preterm birth. PMID- 17127386 TI - Protein i: interference at protein level by intrabodies. AB - Immunoglobulin molecules have long been used in biomedical research as in vitro tools for identification, purification and functional manipulation of target proteins. The specificity and diversity of immunoglobulins can be exploited to target a wide range of intracellular proteins by expressing them in vivo. Such antibody molecules, which are expressed intracellularly and directed to defined sub-cellular compartments, are termed 'intrabodies'. They represent a new and versatile tool that has the potential to manipulate diverse biological processes. Gene knockout, antisense and small interfering RNA knockdown techniques, are employed to characterize the function of many gene products and to validate these gene products as potential drug targets for novel therapeutics, but they have their limitations. The experimental results to date suggest that intrabodies represent a powerful alternative to modulate protein function and analyze its effects. Moreover, they can also be utilized to target specific domains of a particular target protein. Even though their clinical application might take a long time, they can play an important role for target identification and validation in functional genomics and/or proteomics. Here we review the latest advances in the concept, construction and applications of Intrabodies. PMID- 17127387 TI - Phytoplasmas: genetics, diagnosis and relationships with the plant and insect host. AB - Phytoplasmas cannot be cultivated in vitro, and remain the most poorly understood plant pathogens. Despite this limitation, the investigation of their nature with the aid of modern tools has produced noteworthy results during the last 20 years. Using biochemical and molecular approaches, the phylogeny of the phytoplasmas has been described, their chromosomal and extrachromosomal components are being studied, and information on the localization, movement, and metabolic interference occurring in their insect and plant hosts accumulated. At the same time, the application of the new findings in phylogeny and genetics has aided the development of powerful diagnostic tools that have improved the ability to manage diseases which are induced by phytoplasmas. PMID- 17127388 TI - Acid sensing ion channels--novel therapeutic targets for ischemic brain injury. AB - Ischemic stroke is a leading cause of death and long-term disability in the United States. Unfortunately there is no effective therapeutic intervention other than the use of thrombolytics, which has a limited therapeutic time window of approximately 3 h and the potential side effect of intracranial hemorrhage. The absence of neuroprotective therapy is particularly apparent following the failure of multiple clinical trials using glutamate antagonists as therapeutic agents. Understanding the detailed biochemical changes associated with brain ischemia and the cellular mechanisms involved in ischemic brain injury are critical for establishing new and effective neuroprotective strategy. Dramatically decreased tissue pH, or acidosis, is a common feature of ischemic brain, and has been suggested to play a role in neuronal injury. However, the detailed cellular and molecular mechanisms of such acid induced injury remain elusive. The recent finding that acidosis activates a distinct family of cation channels, the acid sensing ion channels (ASICs), in both peripheral and central neurons has dramatically changed the landscape of brain ischemia neurochemistry and provided a novel therapeutic target. In CNS neurons, lowering extracellular pH to the level commonly seen in ischemic brain activates inward ASIC currents resulting in membrane depolarization. In the majority of these neurons, ASICs are also permeable to Ca2+. Therefore, activation of these channels induces an increase of [Ca2+]i. Incubation of neurons with acidic solutions reproduces Ca2+-dependent neuronal injury independent of glutamate receptor activation. The acid-induced currents, membrane depolarization, [Ca2+]i increase, and neuronal injury can be inhibited by the blockade of ASIC1a. In focal ischemia, ASIC1a blockade, or ASIC1a gene knockout both protect brain from injury. The blockers of ASIC1a also demonstrate a prolonged therapeutic time window, beyond that of the glutamate antagonists. Thus, Ca2+-permeable ASIC1a may represent a novel therapeutic target for ischemic brain injury. PMID- 17127389 TI - Hormone treatment enhances WT1 activation of Renilla luciferase constructs in LNCaP cells. AB - The zinc finger transcription factor, WT1, regulates many growth control genes, repressing or activating transcription depending on the gene and cell type. Based on earlier analyses of the effect of WT1 on androgen responsive genes, we hypothesized that there may be an interaction between the androgen signaling pathway and WT1, such that the commonly used Renilla luciferase control vectors were activated in LNCaP prostate cancer cells. Using cotransfection assays we tested the effects of WT1 and/or the androgen analog, R1881, on two Renilla luciferase vectors, pRL-SV40 and the promoter-less pRL-null. To determine whether the zinc finger DNA binding domain was required, the zinc finger mutant DDS-WT1 (R394W) was tested; but it had no significant effect on the Renilla luciferase vectors. To determine whether the androgen signaling pathway was required, WT1 was co-transfected with Renilla vectors in cells with varied hormone responsiveness. The WT1 effect on pRL-null varied from no significant effect in 293 and PC3 cells to very strong enhancement in LNCaP cells treated with 5 nM R1881. Overall, these results suggest that hormone enhanced WT1 mediated activation of Renilla luciferase and that these interactions require an intact WT1 zinc finger DNA binding domain. PMID- 17127390 TI - Geminin in embryonic development: coordinating transcription and the cell cycle during differentiation. AB - Geminin was initially characterized as a bifunctional protein with roles in regulating the fidelity of DNA replication and in controlling cell fate during embryonic nervous system formation. More recently, Geminin's roles have expanded, encompassing regulation of cell proliferation and differentiation during retinogenesis, control of Hox transcription factor function during vertebrate axial patterning, and regulation of the timing of neuronal differentiation. Geminin interacts with homeodomain-containing transcription factors and with protein complexes that regulate chromatin structure, including Polycomb complexes and the catalytic subunits of the SWI/SNF chromatin remodeling complex, Brg1 and Brahma. Activities for Geminin in coordinating cellular events at the transition from proliferation to differentiation have recently emerged in multiple developmental contexts. This review will summarize Geminin's increasingly diverse roles as a developmental regulatory molecule. PMID- 17127391 TI - Myocardial extracellular matrix remodeling in ischemic heart failure. AB - Left ventricular (LV) remodeling is a process whereby structural alterations attempt to compensate altered hemodynamic load. In the chronic setting this process becomes maladaptive, self-sustaining and is associated with worsened survival. The extracellular matrix (ECM) of the heart, once believed an inert scaffold for cardiomyocytes, is now known to play an important role in LV remodeling. The enzyme system primarily responsible for ECM turnover is the matrix metalloproteinases (MMPs), and these enzymes are robustly altered in cardiovascular pathologies, including myocardial infarction (MI) and ischemic heart failure. A cause-and-effect relationship has been established between MMPs and LV remodeling post MI, as MMP inhibition prevents LV dilation and preserves cardiac function in animal models of infarction. In spite of this, initial clinical experience with MMP inhibition post MI has been disappointing. This review examines the structural and functional roles of the myocardial ECM, the evidence for MMP involvement in LV remodeling, and recent investigations into MMPs as prognostic markers and therapeutic targets. PMID- 17127392 TI - Voltage-dependent calcium channels in mammalian spermatozoa revisited. AB - The last few years have seen an explosion in the number of voltage-dependent ion channel sequences detected in sperm and testes. The complex structural paradigm of these channels is now known to include a pore-forming alpha1 subunit(s) whose electrophysiological properties are modulated by an intracellular beta subunit, a disulfide-linked complex of a membrane-spanning delta subunit with an extracellular alpha2 subunit, and a transmembrane gamma subunit. Many of these are alternatively spliced. Furthermore, the known number of genes coding each subtype has expanded significantly (10 alpha1, 4 beta, 4 alpha2delta, 8 gamma). Recently, the CatSper gene family has been characterized based on similarity to the voltage-dependent calcium channel alpha1 subunit. From among this multiplicity, a wide cross-section is active in sperm, including many splice variants. For example, expression of the various alpha1 subunits appears strictly localized in discrete domains of mature sperm, and seems to control distinct physiological roles such as cellular signaling pathways. These include alpha1 alternative splicing variants that are regulated by ions passed by channels in developing sperm. Various combinations of ion channel sequence variants have been studies in research models and in a variety of human diseases, including male infertility. For example, rats that are genetically resistant to testes damage by lead seem to respond to lead ions by increasing alpha1 alternative splicing. In contrast, in varicocele-associated male infertility, the outcome from surgical correction correlates with suppression of alpha1 alternative splicing, Ion channel blockers remain attractive model contraceptive drugs because of their ability to modulate cholesterol levels. However, the large number of sperm ion channel variants shared with other cell types make ion channels less attractive targets for male contraceptive development than a few years ago. In this review, the genetics, structure and function of voltage-dependent calcium channels and related CatSper molecules will be discussed, and several practical clinical applications associated with these channels will be reported. PMID- 17127394 TI - Src kinases in G-CSF receptor signaling. AB - The Granulocyte Colony-Stimulating Factor (G-CSF) receptor, a member of the hematopoietin cytokine receptor superfamily, functions as a homodimer and requires the recruitment of cytosolic protein tyrosine kinases (PTKs) to transduce its signal. At least two cytosolic PTKs are primarily involved: Jak2, a member of the Janus family, and Lyn, a member of the Src family. Through poorly understood mechanisms, these kinases functionally interact with the G-CSF receptor. Jak2 primarily enlists members of the signal transducer and activator of transcription (STAT) family and Lyn phosphorylates a number of adaptor molecules, which link the G-CSF receptor to phosphatidylinositol (PI) 3'-kinase and extracellular signal-regulated kinases (Erk) pathways. This review presents evidence that the Src kinases play a major role in the pathways of G-CSF-mediated proliferation, survival, and differentiation. Identification of Src-dependent pathways provides drug targets useful in the treatment of myeloid leukemias. PMID- 17127395 TI - The dual personalities of matrix metalloproteinases in inflammation. AB - Collagen, gelatin, elastin, fibronectin, proteoglycans and vitronectin are just a few proteins which form the "mesh" that holds a multicellular organism together. The matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that degrade the extracellular matrix. Over several decades it has been clearly established that MMPs are the key molecules associated with matrix remodeling. The remodeling of this matrix is important for physiological and pathological processes such as pregnancy, wound repair, cancer and arthritis. The identification of new non-matrix MMP substrates involved in inflammation, highlights the diverse role of MMPs. These enzymes can enhance leukocyte invasion and regulate the inflammatory activity of serine proteases, cytokines and chemokines. Interestingly, the MMP family appears to have a "dual personality" in that several MMPs such as MMP-2 and -9 can favour either anti- or pro inflammatory action, respectively. The extent of this dual functionality of MMPs is yet to be realized. Elucidating these processes may assist in the development of drugs for the treatment of inflammatory diseases such as arthritis, cancer and chronic wounds. PMID- 17127396 TI - Crack cocaine: effect modifier of RNA viral load and CD4 count in HIV infected African American women. AB - This study reports on the role of cocaine as effect modifier of the association of CD4+ cell counts and RNA viral load. HIV-1 seropositive (n = 80) and seronegative (n = 42) African American women (AAW) crack cocaine smokers were recruited. Increasing cocaine use, based on self-reports and laboratory values, significantly exacerbates the immunopathology of HIV-1 in a dose-response manner, confirmed by a non-linear drop in CD4+ cell number for a given viral load in HIV+ AAW. This report supports a view of deleterious effects due to cocaine use in humans. PMID- 17127393 TI - Roles of gap junctions and hemichannels in bone cell functions and in signal transmission of mechanical stress. AB - Gap junctions formed by connexins (Cx) play an important role in transmitting signals between bone cells such as osteoblasts and osteoclasts, cells responsible for bone formation and bone remodeling, respectively. Gap junction intercellular communication (GJIC) has been demonstrated to mediate the process of osteoblast differentiation and bone formation. Furthermore, GJIC propagates Ca2+ signaling, conveys anabolic effects of hormones and growth factors, and regulates gene transcription of osteoblast differentiation markers. GJIC is also implicated to regulate osteoclast formation, survival and apoptosis. Compared with other bone cells, the most abundant type are osteocytes, which express large amounts of connexins. Mechanosensing osteocytes connect and form gap junctions with themselves and other cells only through the tips of their dendritic processes, a relatively small percent of the total cell surface area compared to other cells. Recent studies show that in addition to gap junctions, osteoblasts and osteocytes express functional hemichannels, the un-opposed halves of gap junction channels. Hemichannels are localized at the cell surface and function independently of gap junctions. Hemichannels in osteocytes mediate the immediate release of prostaglandins in response to mechanical stress. The major challenges remaining in the field are how the functions of these two types of channels are coordinated in bone cells and what the asserted, distinct effects of these channels are on bone formation and remodeling processes, and on conveying signals elicited by mechanical loading. PMID- 17127397 TI - HTLV-1 Tax and adult T-cell leukemia. AB - Human T-lymphotropic virus type I (HTLV-1) is the etiological agent of adult T cell leukemia/lymphoma (ATL) and HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP). HTLV-1 viral transactivator/oncoprotein, Tax, activates viral transcription and usurps regulatory mechanisms that are critical for cell growth and division to facilitate viral replication. The effects that Tax exerts on cells include potent NF-k B activation, cell cycle perturbation and cell transformation. How Tax influences ATL development is incompletely understood at present. While Tax-expression is needed at the early stages of cellular transformation, at later times most ATL cells do not express tax; therefore, genetic and epigenetic changes in HTLV-1-infected cells are believed to play an important role in the etiology of ATL. This review attempts to integrate recent literature on the biological activities of Tax and the properties of HTLV-1 transformed T-cells and ATL cells, and speculate on what cellular changes may collaborate with Tax to effect cell transformation and ATL development. PMID- 17127398 TI - Immune response to retroviral infections of the brain. AB - Various neurological manifestations of retroviral infections have been reported, including peripheral neuropathy, encephalopathy and neuronal degeneration. After penetration into the central nervous system (CNS) the invading retroviruses meet a unique immunological situation that differs significantly from that in the periphery. Due to the blood-brain barrier with its general access restrictions peripheral T-cells, monocytes and B-cells are only "guests" in the brain; instead the immune balance is shifted in favour of the local innate immunity with microglia, astrocytes, cytokines/chemokines and complement forming the dominating defence network. The present article focuses on the most important retroviral infections and highlights the immunological aspects of the neuropathogenesis induced by selected retroviruses. These aspects include: (i) local and infiltrated immune cells as targets of retroviral infection; (ii) stimulation of the cerebral immunity network by retroviruses and subsequent steps of antiviral defence; and (iii) immune activation products as potential contributors to neural damage in the sensitive brain tissue. PMID- 17127399 TI - Cell dynamics and immune response to BLV infection: a unifying model. AB - Bovine Leukemia virus (BLV) is the natural etiological agent of a lymphoproliferative disease in cattle. BLV can also be transmitted experimentally to a related ruminant species, sheep, in which the pathogenesis is more acute. Although both susceptible species develop a strong anti-viral immune response, the virus persists indefinitely throughout life, apparently at a transcriptionally silent stage, at least in a proportion of infected cells. Soon after infection, these humoral and cytotoxic activities very efficiently abolish the viral replicative cycle, permitting only mitotic expansion of provirus carrying cells. Short term cultures of these infected cells initially indicated that viral expression protects against spontaneous apoptosis, suggesting that leukemia is a process of accumulation of long-lived cells. This conclusion was recently reconsidered following in vivo dynamic studies based on perfusions of nucleoside (bromodeoxyuridine) or fluorescent protein markers (CFSE). In sheep, the turnover rate of infected cells is increased, suggesting that a permanent clearance process is exerted by the immune system. Lymphocyte trafficking from and to the secondary lymphoid organs is a key component in the maintenance of cell homeostasis. The net outcome of the immune selective pressure is that only cells in which the virus is transcriptionally silenced survive and accumulate, ultimately leading to lymphocytosis. Activation of viral and/or cellular expression in this silent reservoir with deacetylase inhibitors causes the collapse of the proviral loads. In other words, modulation of viral expression appears to be curative in lymphocytic sheep, an approach that might also be efficient in patients infected with the related Human T-lymphotropic virus type 1. In summary, a dynamic interplay between BLV and the host immune response modulates a complex equilibrium between (i) viral expression driving (or) favoring proliferation and (ii) viral silencing preventing apoptosis. As conclusion, we propose a hypothetical model unifying all these mechanisms. PMID- 17127400 TI - Immune response to maedi-visna virus. AB - The ovine maedi-visna virus (MVV) was the first lentivirus to be isolated and characterized 1957 in Iceland. MVV leads to a life-long, persistent infection with slow development of lesions in the lung and the central nervous system (CNS). The main target cells of MVV are of the monocyte/macrophage lineage and it does not infect T-lymphocytes or cause immune suppression like human immune deficiency virus (HIV). In spite of a fairly good immune response, including both neutralizing antibodies and cytotoxic T lymphocytes, the virus persists in the host and establishes a life-long infection. There are strong indications that the pathological lesions are immune-mediated and vaccination attempts have not only failed to induce sterile immunity but have occasionally caused increased viremia and more severe disease. PMID- 17127401 TI - Immune control and prevention of chronic Friend retrovirus infection. AB - T cells are critical to control acute infection of a host with retroviruses but they are usually unable to prevent the development of chronic infections. This review summarizes studies from the Friend virus mouse model that reveal some of the mechanisms by which T cells control chronic retroviral infection, and also reveal why these responses ultimately fail to fully eradicate infection. Also summarized are findings from vaccine studies demonstrating the immunological requirements for the prevention of chronic retroviral infection. The implications of these findings for chronic infections in humans are discussed. PMID- 17127402 TI - Junction interaction in the seminiferous epithelium: regulatory roles of connexin based gap junction. AB - Anchoring junction, tight junction (TJ), and gap junction (GJ) constitute three major junction types in mammalian testes. Connexin is the well-studied GJ protein. It forms the building block of connexon, which is composed of six connexin units. Connexon forms the functional GJ when pairing with counter connexon from neighboring cells. In the testis, at least eleven connexins are associated with the Sertoli and germ cells of the seminiferous epithelium and the Leydig cells of the interstitium, modulating spermatogenesis and steroidogenesis, respectively. Significantly, connexins are recently speculated to act as regulators of other junctions in the testes using pan-connexin peptide model. This demonstrates that the loss of connexin function leads to a preferential degradation of occludin-based TJ, but not N-cadherin-based adherens junction (AJ), in the testis, despite the intermingled relationship of these three junctions at the site of blood-testis barrier. In the clinical aspects, connexins are shown to relate to male infertility and testicular dysfunctions. A panel of molecules and proteins and their associated protein kinases are actively participating in the regulation of connexin-mediated GJ and fine-tuning connexin associated functions in the testis. Herein, we summarize the latest findings of connexins in the testis in the aspects of fertility, and testicular diseases, with emphasis on the unexplored roles of connexins in regulating other junction types. This can shed light on future studies in implicating the putative roles of connexins in the physiological functions of reproduction and the clinical aspects of male infertility. In addition, understanding the roles of connexins can advance the diagnosis and treatment of testicular dysfunction and infertility. PMID- 17127403 TI - Mapping the future of common diseases: lessons from psoriasis. AB - Psoriasis (OMIM*177900) is a common, chronic, hyperproliferative inflammatory disorder of the skin affecting approximately 2% of Caucasians. Despite the prevalence of psoriasis in general population, significant differences in the incidence among Japanese, Eskimos, West Africans, north American blacks and American natives are well known. The cause for these variations are likely to be both genetic and environmental. Independent genomewide scans have suggested the involvement of a large number of chromosomal regions (loci), but so far only poor susceptibility genes have been suggested. We discuss genetic basis of the disease, results and interpretations of relevant studies, with particular regard to study design and future perspectives. Indeed to date, mapping genes which contribute to complex diseases is one of the major challenge in the post-genomic era. PMID- 17127404 TI - Cyclooxygenases and lipoxygenases in prostate and breast cancers. AB - Oxidative metabolism of polyunsaturated fatty acids through cyclooxygenases or lipoxygenases can generate various lipid peroxides and bioactive lipids, and regulate cellular proliferation, apoptosis, differentiation and senescence. The role of the second cyclooxygenase isoform (COX-2) has been demonstrated in a number of studies and is regarded as a promising target for chemoprevention and treatment. The involvement for lipoxygenases in tumor initiation and progression has been implicated in several studies but remains controversial. Among the many members of lipoxygenase family, both tumor promoting and suppressing activities have been described. For example, 15-lipoxygenase-1 has been implicated as a tumor promoter in prostate cancer, but it suppresses colon cancer. In this review, the role of cyclooxygenases and lipoxygenases in cancer will be described, with the hope of attracting further research to define their functions in cancer. PMID- 17127405 TI - Transgenic expression of human INS gene in Ins1/Ins2 double knockout mice leads to insulin underproduction and diabetes in some male mice. AB - We have generated transgenic mouse lines expressing exclusively a human INS transgene on an Ins1/Ins2 double knockout (mIKO) background. The transgene expression was driven by either a 4000 bp or a 353 bp promoter. These transgenic lines, designated mIKO:INS4000 and mIKO:INS353, were viable and fertile. Determination of the amounts of insulin transcripts and total pancreatic insulin content revealed relative insulin underproduction in both lines, from birth to adulthood. Total pancreatic insulin stores in mIKO:INS4000 and mIKO:INS353 mice represented only about 50% and 27%, respectively, as compared to wild-type mice. Morphometric analysis of pancreas did not show any compensatory beta-cell hyperplasia. The majority of animals in both lines remained normoglycemic throughout their lives. Nevertheless, glucose tolerance tests revealed glucose intolerance in nearly half of mIKO:INS4000 male mice, likely due to impaired insulin secretion detected in those animals. In addition, a small fraction (2-4%) of male mice in both lines spontaneously developed diabetes with very distinct pathophysiological features. Diabetes was never seen in female animals. The diabetes developed by mIKO:INS353 mice was rapidly lethal, accompanied by a dramatic depletion of pancreatic insulin stores whereas the mIKO:INS4000 diabetic animals could live for several months. This suggests a possible link between the structure of the human INS gene promoter and the type of diabetes developed in these lines. PMID- 17127406 TI - Immune response to MMTV infection. AB - Mouse mammary tumor virus (MMTV) has developed a strategy of exploitation of the immune response. It infects dendritic cells and B cells and requires this infection to establish an efficient chronic infection. This allows transmission of infection to the mammary gland, production in milk and infection of the next generation via lactation. The elaborate strategy developed by MMTV utilizes several key elements of the normal immune response. Starting with the infection and activation of dendritic cells and B cells leading to the expression of a viral superantigen followed by professional superantigen-mediated priming of naive polyclonal T cells by dendritic cells and induction of superantigen mediated T cell B cell collaboration results in long-lasting germinal center formation and production of long-lived B cells that can later carry the virus to the mammary gland epithelium. Later in life it can induce transformation of mammary gland epithelium by integrating close to proto-oncogenes leading to their overexpression. Genes encoding proteins of the Wnt-pathway are preferential targets. This review will put these effects in the context of a normal immune response and summarize important facts on MMTV biology. PMID- 17127407 TI - Predicting single nucleotide polymorphisms (SNP) from DNA sequence by support vector machine. AB - Recently, SNP has gained substantial attention as genetic markers and is recognized as a key element in the development of personalized medicine. Computational prediction of SNP can be used as a guide for SNP discovery to reduce the cost and time needed for the development of personalized medicine. We have developed a method for SNP prediction based on support vector machines (SVMs) using different features extracted from the SNP data. Prediction rates of 60.9% was achieved by sequence feature, 59.1% by free-energy feature, 58.1% by GC content feature, 58.0% by melting temperature feature, 56.2% by enthalpy feature, 55.1% by entropy feature and 54.3% by the gene, exon and intron feature. We introduced a new feature, the SNP distribution score that achieved a prediction rate of 77.3%. Thus, the proposed SNP prediction algorithm can be used to in SNP discovery. PMID- 17127408 TI - Corticotropin-releasing hormone and the blood-brain-barrier. AB - Increased blood-brain-barrier (BBB) permeability precedes any clinical or pathologic signs and is critical in the pathogenesis of multiple sclerosis (MS) and brain metastases. CD4+ TH1 cells mediate demyelination in MS, but how they get sensitized and enter the brain to induce brain inflammation remains obscure. TH2 cytokines associated with allergic disorders have recently been implicated in MS, while genes upregulated in MS plaques include the mast cell-specific tryptase, the IgE receptor (Fc-epsilon-RI) and the histamine-1 receptor. Mast cell specific tryptase is elevated in the CSF of MS patients, induces microvascular leakage and stimulates protease-activated receptors (PAR), leading to widespread inflammation. BBB permeability, MS and brain metastases appear to worsen in response to acute stress that leads to the local release of corticotropin-releasing hormone (CRH), which activates brain mast cells to selectively release IL-6, IL-8 and vascular endothelial growth factor (VEGF). Acute stress increases BBB permeability that is dependent on CRH and mast cells. Acute stress shortens the time of onset of experimental alleric encephalomyelitis (EAE) that does not develop in W/W mast cell deficient or CRH -/- mice. Brain mast cell inhibition and CRHR antagonists offer novel therapeutic possibilities. PMID- 17127409 TI - Cdt1 and geminin: role during cell cycle progression and DNA damage in higher eukaryotes. AB - DNA replication in eukaryotic cells must be strictly regulated to ensure that the entire genome is duplicated only once in each cell cycle. For this purpose, the initiation of DNA replication is controlled by the "licensing" reaction, which is established by the formation of a pre-replicative complex (pre-RC) with the sequential assembly of the origin recognition complex (ORC), Cdc6, Cdt1 and Mcm2 7 onto origin regions. Among these, Cdt1 is likely the most important target for regulating licensing in higher eukaryotic cells, since illegitimate accumulation of Cdt1 causes multiple rounds of DNA replication without an intervening mitosis. Cdt1 is regulated over the course of the cell cycle mainly by the controlled expression of an inhibitor protein, geminin, and the level of Cdt1 periodically fluctuates due to ubiquitination and proteolysis. While the expression of geminin from S phase to metaphase of mitosis prevents licensing, Cdt1 accumulates from M to G1 phases and is degraded at the onset of S phase. Furthermore, Cdt1 is also proteolyzed in G1 phase in response to DNA damage, presumably providing a new checkpoint control. PMID- 17127410 TI - Hydrogen peroxide selectively increases TREK-2 currents via myosin light chain kinases. AB - Two-pore domain K+ (K2P) channels play a critical role in cellular responses to various stimuli, such as stretch or changes in pH and are considered to be important in pathological responses such as apoptosis and tumorigenesis. We investigated effects of H2O2 on various K2P channels expressed in CHO cells. Application of H2O2 did not affect TASK-1, TASK-3, TRAAK currents, but specifically increased TREK-2 currents recorded using a nystatin perforated whole cell technique. The H2O2-induced activation of TREK-2 currents was also observed at single channel levels in cell-attached patches, and the effect was reversed by the reducing agent, dithiothreitol. In contrast, TREK-2 currents recorded using ruptured whole cell technique or single channel recording in inside-out patches were not affected by H2O2. Furthermore, direct application of 5,5'-dithiobis-(2 nitrobenzoic acid) inhibited TREK-2, suggesting that the H2O2-induced activation does not result from direct oxidation of TREK-2 proteins. Among the cell signaling agents, myosin light chain kinase (MLCK) inhibitors significantly inhibited the H2O2-induced activation of TREK-2 currents. These results suggest that TREK-2 channels have a potential to play a specific role in cellular responses to reactive oxygen species and that MLCK activation is involved in this process. PMID- 17127411 TI - Aberrant Gi protein coupled receptor-mediated cell survival signaling in rheumatoid arthritis B cell lines. AB - Sphingosine 1-phosphate (S1P) is a pleiotropic bioactive lipid that transmits potent signals through a family of G protein coupled receptors with resultant anti-apoptotic and pro-angiogenic effects. We have recently reported that lymphoblastoid B cell lines (LCLs) from rheumatoid arthritis (RA) patients are resistant to Fas-mediated cell death due to over-production of S1P, secondary to over-activity of sphingosine kinase-1 (SphK1). Here we investigated the signaling events that S1P triggers in those cells. Our results show that RA-derived LCLs display increased constitutive enzymatic activity of phosphatidylinositol 3 kinase (PI3K). Incubation of LCLs with a PI3K inhibitor wortmannin reversed PI3K over-activity and the resistance to Fas-mediated cell death. Incubation of RA LCLs with nanomolar concentration of S1P triggered exaggerated activation of both SphK and PI3K in RA LCLs compared to control cells. PI3K was mapped upstream of SphK, since wortmannin could block SphK activation by S1P. S1P signaling effect could be blocked by the Gi/G0 protein inhibitor, pertussis toxin and by an inhibitor of S1P-receptor interaction, suramin. S1P receptor expression levels did not appear to be the cause of disparate S1P-triggered signaling, since LCLs from RA patients and their healthy twin controls did not show statistically significant differences in the expression levels of the five known S1P receptors, as determined by quantitative real time reverse transcription-polymerase chain reaction analyses. Thus, we conclude that Fas death signaling aberration in RA LCLs is caused by extracellular S1P, which triggers PI3K-dependent SphK over activity through a Gi protein-coupled receptor-mediated signaling cascade. PMID- 17127412 TI - Identification of ICIS-1, a new protein involved in cilia stability. AB - Cilia are specialized organelles that exert critical functions in numerous organisms, including that of cell motility, fluid transport and protozoan locomotion. Ciliary architecture and function strictly depend on basal body formation, migration and axoneme elongation. Numerous ultrastructural studies have been undertaken in different species to elucidate the process of ciliogenesis. Recent analyses have led to identification of genes specifically expressed in ciliated organisms, but most proteins involved in ciliogenesis remain uncharacterized. Using human nasal epithelial cells capable of ciliary differentiation in vitro, differential display was carried out to identify new proteins associated with ciliogenesis. We isolated a new gene, ICIS-1 (Involved in CIlia Stability-1), upregulated during mucociliary differentiation. This gene is localized within the TGF-beta1 promoter and is ubiquitously expressed in human tissues. Functional analyses of gene expression inhibition by RNA interference in Paramecium tetraurelia indicated that the ICIS-1 homologue interfered with cilia stability or formation. These findings demonstrate that ICIS-1 is a new protein associated with ciliated cells and potentially related to cilia stability. PMID- 17127413 TI - Craniofacial, skeletal, and cardiac defects associated with altered embryonic murine Zic3 expression following targeted insertion of a PGK-NEO cassette. AB - Mutation in ZIC3 (OMIM #306955), a zinc finger transcription factor, causes heterotaxy (situs ambiguus) or isolated congenital heart defects in humans. Mice bearing a null mutation in Zic3 have left-right patterning defects with associated cardiovascular, vertebra/rib, and central nervous system malformations. Although XZic3 is thought to play a critical role in Xenopus neural crest development, no defects in tissues derived from neural crest are apparent in adult Zic3(null) mice. In this study we have characterized the effect of a PGK-neo cassette insertion 5' of the Zic3 locus. The Zic3 transcript in this new allele is up-regulated in ES cells and in E9.0 embryos, but no ectopic expression was detected. Unlike the Zic3(null) mutation in which only 20% of mutant animals survive to adulthood, there was no evidence of excess fetal death caused by the Zic3(neo) allele. Zic3(neo) mutant mice exhibited hemifacial microsomia, asymmetric low set ears, axial skeletal defects, kyphosis and scoliosis; a combination of defects which mimics Goldenhar Syndrome. Some Zic3(neo) mice had evidence of left-right axis patterning defects, but cardiac malformation was much less common than in the Zic3(null) mutants. A six-week old hemizygous mouse was found to have thoraco-cervical ectopia cordis, an extremely rare congenital malformation in humans and for which there is no precedent in a mouse model. PMID- 17127414 TI - Hepatic differentiation and transcriptional profile of the mouse liver epithelial progenitor cells (LEPCs) under the induction of sodium butyrate. AB - The liver regenerates by progenitor cells when it is damaged in chronic liver diseases and extensive damage. The progenitor cells, also termed "oval cells" according to their morphological traits, can differentiate into hepatocytes and bile duct cells in vivo. To better understand the transcriptional pattern that accompanies the hepatic differentiation of oval cells, we applied cDNA microarray to analyze the oval cell-derived liver epithelial progenitor cells (LEPCs) during in vitro induced differentiation. Upon exposure to sodium butyrate, a histone deacetylase inhibitor, cultured LEPCs differentiate and express functional hepatocyte markers albumin, tryptophan 2, 3-dioxygenase and alcohol dehydrogenase. For expression profiling, cells were harvested at 6 h, 12 h, 1 d, 3 d and 7 d after exposure to sodium butyrate. After analyzing the microarray data by SOM clustering, total of 796 differentially regulated genes were grouped into 48 clusters. Consistent with the phenotype change of LEPCs after sodium butyrate treatment, many hepatocyte functional genes are revealed by analyzing the clusters containing genes up-regulated through all the time points. The clusters, containing down-regulated genes immediately after the induction, are also analyzed. The microarray data was validated by analyzing the expression of selected genes by quantitative real-time PCR. A set of genes expressed synergistically in these clusters may play a central role during the process of differentiation. Sodium butyrate decreases cyclin B1 and Cdk4 expression, which would be associated with LEPCs growth arrest shortly after treatment. Bmi1, a polycomb group protein, is also down-regulated immediately after treatment and remains at a low level during the induction. These findings highlight the key molecular mechanisms by which sodium butyrate, mediates its effects on cell growth arrest and induction of differentiation. In conclusion, our data reflect a global view of gene expression during hepatic differentiation of LEPCs induced by sodium butyrate. PMID- 17127415 TI - Identification of genes differentially regulated by transcription factor, AP 2delta. AB - In an effort to better understand the transcriptional program regulated by transcription factor, AP-2delta, we used cDNA microarray to evaluate the relative expression of human genes in AD293 cells by exogenous expression of AP-2delta. Microarray showed 29 genes that were up-regulated and 39 genes with a down regulated expression pattern. Among the identified genes were those encoding transcription factors, signal transduction molecules, kinases, as well as genes regulating cell growth, differentiation, and cell adhesion, a set of genes suggested to be regulated by AP-2. The results of microarray was validated for some regulated genes by real-time PCR analysis. These observations shed novel insight into the mechanism of AP-2delta action and provide a range of target genes for further investigation. PMID- 17127416 TI - Genetics of autoreactive B cells. AB - It is now well-accepted that autoimmune diseases develop as a result of interactions between a complex genetic basis and environmental triggers. Autoreactive B cells play a major role in many autoimmune diseases, by secreting autoantibodies or cytokines and/or presenting auto-antigens to T cells. Studies performed with human patients and murine models have accumulated evidence that B cell autoreactivity, or its manifestation as the presence of autoantibodies, are also supported by multiple genetic determinants. These studies will be summarized in this review and presented in a critical perspective of the approaches used to obtain these results, and their significance for our understanding of B-cell tolerance. PMID- 17127417 TI - The role of B cells in animal models of rheumatoid arthritis. AB - Rheumatoid arthritis is a chronic systemic inflammatory autoimmune disease that affects approximately 1% of the population. Recent studies demonstrate a significant improvement in clinical symptoms in patients treated with Rituximab, an anti-CD20 monoclonal antibody that depletes pro-B cells and mature B cells but not plasma cells. These findings indicate that B cells are an important contributor to the pathogenesis of RA. In this review we will examine the role of B cells in several different murine models of RA. There are a number of antibody dependent mechanisms by which B cells support inflammatory processes in the joint. However, there are also antibody-independent mechanisms that involve B cell/T cell collaboration where B cells may modulate autoreactive T cell responses. In addition, B cells may be an important source of cytokines that either stimulate or inhibit autoimmune responses. Understanding the role of B cells in RA will provide new and directed therapeutic approaches to the treatment of disease. PMID- 17127418 TI - Mild hypothermia diminishes oxidative DNA damage and pro-death signaling events after cerebral ischemia: a mechanism for neuroprotection. AB - Mild hypothermia, applied either during or soon after cerebral ischemia, has been shown to confer robust neuroprotection against brain injury in experimental stroke and in patients recovering from cardiac arrest. However, the mechanism underlying hypothermic neuroprotection is not completely understood. In this study, the effect of mild hypothermia on the induction of oxidative DNA damage, an early harmful event during post-ischemic reperfusion that triggers both necrotic and apoptotic cell death in the brain, was studied using the rat model of middle cerebral artery occlusion (MCAO) and reperfusion. Rats were subjected to 2-hr MCAO and reperfusion of various durations up to 3 days. Selective brain hypothermia (33 degrees C) was induced at the onset of ischemia and terminated at the beginning of reperfusion, and this significantly decreased infarct volume 72 hr later. Correlated with this protective effect, intraischemic mild hypothermia markedly attenuated the nuclear accumulations of several oxidative DNA lesions, including 8-oxodG, AP sites, and DNA single-strand breaks, after 2-hr MCAO. Consequently, harmful DNA damage-dependent signaling events, including NAD depletion, p53 activation, and mitochondrial translocation of PUMA and NOXA, were reduced during post-ischemic reperfusion in hypothermia-treated brains. These results suggest that the attenuation of oxidative DNA damage and DNA damage triggered pro-death signaling events may be an important mechanism underlying the neuroprotective effect of mild hypothermia against ischemic brain injury. PMID- 17127419 TI - New insights into sperm-zona pellucida interaction: involvement of sperm lipid rafts. AB - Sperm-zona pellucida (ZP) binding is the first step of gamete interaction. This binding occurs in two sequential steps, starting with the primary binding of acrosome-intact sperm to the ZP followed by the secondary ZP binding of acrosome reacting/reacted sperm. While there are only a few ZP sulfoglycoproteins involved in these binding events, a large number of sperm surface molecules have been shown to possess ZP affinity. In this review, we have given explanations to the existence of these many ZP binding molecules. We have also summarized their origin and the mechanisms of how they are targeted to the sperm surface and acrosome. Recently, we have shown that sperm lipid rafts have affinity for the ZP. A number of ZP binding molecules are also present in sperm lipid rafts. In this review, we have provided an argument that sperm lipid rafts may be the platforms on the sperm surface for ZP interaction. PMID- 17127420 TI - Sjogren's syndrome (SjS)-like disease of mice: the importance of B lymphocytes and autoantibodies. AB - Sjogren's syndrome (SjS) is a systemic autoimmune disease in which an immunological attack against the salivary and lacrimal glands results, respectively, in severe dry mouth and dry eye diseases. Although a CD4+ T lymphocyte population is an integral component in the pathogenesis of SjS, recent studies have focused on the importance the B lymphocyte plays in both the pre clinical and clinical phases of the disease process. To understand the molecular and cellular mechanisms involved in SjS, numerous mouse models that mimic major clinical manifestations of the human disease have been developed. Studies have begun to define the genetics, the nature of the autoimmune response towards the salivary and lacrimal glands, as well as the possible mechanisms for effecting glandular dysfunction, thereby establishing insights to new intervention therapies. Not surprising, the B cell is taking center stage. Here, we present an in-depth discussion of how B cell populations may be involved in orchestrating or determining exocrine gland dysfunction. PMID- 17127421 TI - Aberrant B1 cell trafficking in a murine model for lupus. AB - B lymphocyte chemoattractant (BLC/CXCL13) is ectopically and highly expressed in the target organs such as the thymus and kidney in aged (NZB x NZW)F1 (BWF1) mice, a murine model for SLE. Ectopic expression of BLC/CXCL13 was attributed to mature myeloid DCs infiltrating in the target organs. DCs were also increased in the peripheral blood in aged BWF1 mice and differentiated into BLC/CXCL13 producing DCs in the presence of TNF-alpha or IL-1beta, but not IFN-alpha or IFN gamma. BLC/CXCL13 expression in mature myeloid DCs was confirmed in bone marrow derived-DCs generated in vitro in the presence of GM-CSF and TNF-alpha. B1 cells expressed higher level of CXCR5 and migrated towards BLC/CXCL13 much better than B2 cells. B1 cells failed to home to the peritoneal cavity and preferentially recruited to the target organs in aged BWF1 mice developing lupus nephritis. B1, but not B2 cells possessed a potent antigen presenting activity in allo-MLR and activated autologous thymic CD4 T cells in the presence of IL-2. CXCR5+ CD4 T cells were also increased in aged BWF1 mice and they enhanced IgG production by B1 cells in vitro. These results suggest a possible involvement of aberrant B1 cell trafficking in activation of autoreactive CD4 T cells and autoantibody production in the target organs during the development of lupus, providing a new insight for the pathogenesis of B1 cells in lupus. PMID- 17127422 TI - Luteinizing hormone receptors expression in cumulus cells closely related to mouse oocyte meiotic maturation. AB - In the process of oocyte maturation, gonadotrophins are believed as main stimulators for oocyte meiosis resumption. However, which gonadotrophin (i.e. FSH or LH) is the key hormone in this process is still unknown. This study indicated a close relationship between LH and FSH on activating meiotic maturation of oocyte in vitro. FSH efficiently induced oocyte meiosis at concentration of 50 IU/L, while LH alone had no effect on oocyte meiotic initiation. Using RT-PCR and in situ hybridization, a tight corelationship between FSH-stimulated oocyte meiotic resumption and LHR mRNA expression in cumulus cells was found. LHR mRNA was present in cumulus cells of oocyte-cumulus cell complexes. Except the expression of LHRs was present in cumulus cells surrounding all maturing oocytes, low level expression was also detected in cells associated with oocytes that were still at GV stage. Its expression was enhanced by FSH stimulation before oocyte maturation. However, LHRs did not express in cumulus cells associated with oocytes that were completely arrested at GV stage by IBMX. Furthermore, increased progesterone concentration was found in the medium in which CEOs were cultured with FSH and LH, but not in those with FSH or LH alone. LHR expression in cumulus cells increases with time in culture, and the levels of expression are enhanced in the presence of FSH. Oocyte meiotic resumption may create conditions favorable for LHR expression. LHR expression may be considered as a potential marker for oocytes maturation initiation. PMID- 17127423 TI - Multisite phosphorylation of the cAMP response element-binding protein (CREB) by a diversity of protein kinases. AB - The prevailing view of stimulus-induced activation of the transcription factor cAMP response element-binding protein (CREB) presumes phosphorylation at serine 133. Although, phosphorylation of this residue seems to be necessary, it is not sufficient to trigger CREB-driven transcription, indicating that other phosphoserine-133-independent mechanisms are required for full activation of CREB. One of these additional regulatory mechanisms influencing the transcriptional state of CREB may involve multiple phosphorylation events on other phosphoacceptor sites in the protein. This review focuses on the phosphorylation modifications of CREB by distinct protein kinases and discusses the possible implications for the function of CREB. PMID- 17127424 TI - Peptide vaccines against cancer, infectious diseases, and conception. AB - The concept of peptide vaccines is based on identification and chemical synthesis of B-cell and T-cell epitopes which are immunodominant and can induce specific immune responses. B-cell epitope of a target molecule can be coupled to a promiscuous T-cell epitope to make it immunogenic. Our increased understanding of antigen recognition at molecular level has resulted in the development of rationally designed peptide vaccines. The relative ease of construction and production, chemical stability, and lack of oncogenic or infectious potential has made the peptides attractive vaccine candidates. However, several obstacles limit the widespread usefulness of peptide vaccines. These include their low immunogenicity, need for a better adjuvant and carrier, and reliable and simple assays to measure T-cell response. Nonetheless, current efforts are defying these limitations and many promising discoveries are making their way to improve this approach. The peptide vaccines against various cancers have undergone phase I and phase II clinical trials with successful immunological clinical outcome. The peptide vaccination is being examined both for palliative and prophylactic immunotherapy. The current status of many peptide vaccines which are being developed against cancer, infectious diseases, and conception is discussed in this review. PMID- 17127425 TI - Identification of differentially expressed genes in multiple microarray experiments using discrete fourier transform. AB - Research in the post-genome sequence era has been shifting towards a functional understanding of the roles and relationships between different genes in different conditions. While the advances in genetic expression profiling techniques including microarrays enable detailed and genome-scale measurements, the extraction of meaningful information from large datasets remains a challenging task. Here, we propose a novel method of generating gene differential expression profiles such that gene expression values from one dataset can be directly compared with those of another dataset. A simplified Discrete Fourier Transform is applied to interposed gene expression values, thereby generating the 'spectra' for a pair of conditions. Using this technique, differentially expressed genes produce higher amplitudes at the Nyquist Frequency. By measuring the phase of the 'spectra' generated, the over- and under-expressed nature of the genes can be identified. This method was validated using two sets of GeneChip array data, one from prostate cancer related dataset and the other from macular degeneration related dataset. The genes identified as differentially expressed by our method were found to be similar to those published using their preferred methods. Based on our findings, the proposed DFT method could be used efficiently in identifying differentially expressed genes from multiple-array experiments from two different conditions. PMID- 17127426 TI - Impaired DNA repair via the base-excision repair pathway after focal ischemic brain injury: a protein phosphorylation-dependent mechanism reversed by hypothermic neuroprotection. AB - Cerebral ischemia and reperfusion induces rapid accumulation of oxidative DNA lesions in the brain, which, if not repaired promptly, may trigger cell death. The base-excision repair (BER) pathway is the main mechanism employed by neurons to repair various types of oxidative DNA damage. Recent studies have suggested that the cellular activity of BER is highly regulated (up- or down-regulated) after ischemic brain injury, and this regulation may contribute to the outcome of cell injury. The mechanism through which cellular BER is regulated in response to neuronal injury is currently poorly understood. In the present study, we have examined BER regulation in the rat model of focal ischemic brain injury induced by 2 hr of middle cerebral artery occlusion and 0-72 hr of reperfusion. As determined using cerebral nuclear extracts, focal ischemia resulted in a marked reduction in BER activities, including the overall BER activity, AP endonuclease activity and DNA polymerase-beta activity, indicating functional impairment of the BER pathway. BER reduction occurred as early as 0.5 hr after the onset of reperfusion. Thereafter, BER activity failed to recover, and there were persistent accumulations of apurinic/apyrimidinic abasic sites and DNA single strand breaks in ischemic tissues. The reduction in BER during the early reperfusion phase (less than 6 hr) was not accompanied by any alterations in the levels of essential BER enzymes in brain extracts. However, increased serine- and threonine-specific phosphorylation was detected for both AP endonuclease and DNA polymerase-beta after ischemia, with the time course of serine phosphorylation closely correlated to that of changes in BER activity. Furthermore, dephosphorylation of nuclear extracts with alkaline phosphatase largely restored AP endonuclease and DNA polymerase-beta activities. Taking advantage of the neuroprotective effect of mild hypothermia (33 degrees C), which was induced in the brain during the first 2 hr of reperfusion, we found that the post-ischemic suppression of BER activity is a reversible event. Hypothermic treatment diminished the serine-specific phosphorylation of AP endonuclease and DNA polymerase-beta, promoted BER activities, and attenuated the levels of oxidative DNA lesions after ischemia. These results suggest that the functional impairment of the BER pathway after severe focal cerebral ischemia is due to the loss-of function post-translational modifications of repair enzymes. Further investigations elucidating the precise mechanism underlying the post translational regulation of BER enzymes may lead to novel therapeutic strategies for cerebral ischemia. PMID- 17127427 TI - NAD+ and NADH in brain functions, brain diseases and brain aging. AB - Numerous studies have suggested that NAD+ and NADH mediate multiple major biological processes, including calcium homeostasis, energy metabolism, mitochondrial functions, cell death and aging. In particular, NAD+ and NADH have emerged as novel, fundamental regulators of calcium homeostasis. It appears that most of the components in the metabolic pathways of NAD+ and NADH, including poly(ADP-ribose), ADP-ribose, cyclic ADP-ribose, O-acetyl-ADP-ribose, nicotinamide and kynurenine, can produce significant biological effects. This exquisiteness of NAD+ and NADH metabolism could epitomize the exquisiteness of life, through which we may grasp the intrinsic harmony life has evolved to produce. The exquisiteness also suggests a central regulatory role of NAD+ and NADH in life. It is tempted to propose that NAD+ and NADH, together with ATP and Ca2+, constitute a Central Regulatory Network of life. Increasing evidence has also suggested that NAD+ and NADH play important roles in multiple biological processes in brains, such as neurotransmission and learning and memory. NAD+ and NADH may also mediate brain aging and the tissue damage in various brain illnesses. Our latest studies have suggested that NADH can be transported across the plasma membranes of astrocytes, and that NAD+ administration can markedly decrease ischemic brain injury. Based on this information, it is proposed that NAD+ and NADH are fundamental mediators of brain functions, brain senescence and multiple brain diseases. Because numerous properties of NAD+ and NADH remain unclear, future studies regarding NAD+ and NADH may expose some fundamental mechanisms underlying brain functions, brain pathologies and brain aging. PMID- 17127428 TI - Role of autoantibodies in type 1 diabetes. AB - Type 1A, the immune mediated form of diabetes, is a relatively common disorder that develops in genetically susceptible individuals. The disease is associated with a series of anti-islet autoantibodies and the autoantibodies can be present for years prior to the onset of hyperglycemia. In general it is thought that type 1A diabetes is T cell mediated, but there is evidence from studies in the NOD mouse model that antibodies and B-lymphocytes contribute to pathogenesis. In man evidence is lacking that transplacental passage of anti-islet antibodies increases disease risk. Well characterized, high throughput autoantibody assays (tested in a series of international workshops) are now available, and are the mainstays of prediction of type 1A diabetes, diagnosis of the immune mediated form of diabetes, and are important for the design of trials for the prevention of type 1A diabetes. In addition to anti-islet autoantibodies, patients with type 1A diabetes develop a series of associated autoimmune disorders that are usually detected with screening for additional autoantibodies (e.g. anti-thyroid, anti transglutaminase, anti-21 hydroxylase, anti-parietal cell). At present it is possible to predict the development of type 1A diabetes and prevent the disorder in animal models, but we lack proven therapies for disease prevention in man. The ability to detect specific anti-islet autoantibodies provides the foundation for developing and testing these preventive therapies. PMID- 17127429 TI - Phosphorylation of phosphodiesterase-5 is promoted by a conformational change induced by sildenafil, vardenafil, or tadalafil. AB - Phosphodiesterase-5 (PDE5) inhibitors (sildenafil, vardenafil, or tadalafil) or phosphorylation by cyclic nucleotide-dependent protein kinase causes an apparent conformational change in PDE5, as indicated by a shift in migration on non denaturing PAGE gels and an altered pattern of tryptic digestion. Combination of cGMP and a PDE5 inhibitor or phosphorylation does not cause a further gel shift or change in tryptic digest. Phosphorylation of PDE5 is stimulated by inhibitors, and combination of cGMP and inhibitor does not cause further phosphorylation. Dephosphorylation of PDE5 by either purified phosphoprotein phosphatase-1 or -2A catalytic subunit or by a crude phosphatase mixture is not affected by cGMP or inhibitors, suggesting that phosphorylation itself maintains conformational exposure of the phosphorylation site. The combined results imply that cGMP binding to the catalytic site initiates negative feedback control of many cellular cGMP signaling pathways by directly stimulating phosphorylation and activation of PDE5; by exploiting this molecular mechanism, PDE5 inhibitors stimulate their own potencies. PMID- 17127430 TI - BS69, a corepressor interacting with ZHX1, is a bifunctional transcription factor. AB - Corepressor BS69 interacts with ZHX1, a member of the ZHX family having zinc fingers and homeoboxes. In the rat, we have identified four forms of splicing variants, BS69alpha, BS69beta, BS69gamma, and BS69delta. Based on the amino acid sequence, BS69alpha corresponded to the human orthologue. BS69beta and BS69gamma contain a novel 56 amino acid region encoded by the exon 11b of the rat BS69 gene. Both BS69gamma and BS69delta lacked a region encoded by exon 3 of the gene. Although all four variants were ubiquitously expressed in rats, the transcripts having the exon 11b were detected in mice and rats but not in humans. A common C terminal MYND domain of BS69 was required for the interaction with PxLxP motif of ZHX1. Although BS69 was originally found as a corepressor interacting with ZHX1, BS69 was also found to function as a transcriptional activator in HEK293 cells, in which the activation required the MYND domain of BS69. Co-transfection of BS69 with a mutant form of ZHX1, which cannot interact with BS69, led to increase the transcriptional activation of BS69, suggesting that transcriptional activation mediated by BS69 is suppressed by ZHX1. In contrast, BS69 showed transcriptional repression in COS-7 and CV-1 cells and the repression domain was mapped to the N terminus of BS69beta. Both the wild type and mutant form of ZHX1 had no effect on the BS69 repression, suggesting that the repression mediated by BS69 in COS-7 and CV-1 cells may require a cofactor other than ZHX1 in the cells. Therefore, our results suggest that BS69 may function either as a transcriptional repressor or as a transcriptional activator depending on its regulatory partner. PMID- 17127431 TI - Susceptibility to inhalation toxicity of acetaldehyde in Aldh2 knockout mice. AB - In this study, we evaluated the inhalation toxicity of acetaldehyde in Aldh2 KO (Aldh -/-) mice, using pathological method. Male C57BL/6 (Aldh2 +/+) mice and Aldh -/- mice were exposed to atmospheres containing acetaldehyde at levels of 0, 125, and 500 ppm for 24 h/day during 14 days. Although the average blood acetaldehyde concentration of Aldh -/- mice was higher than that of Aldh2 +/+ mice in the acetaldehyde exposure group, observable effects by the acetaldehyde exposure on the lung and liver were not different between wild type and ALDH2 null mice. In Aldh2 -/- mice, the levels of 1) erosion of respiratory epithelium and the subepithelial hemorrhage in nose, 2) hemorrhage in nasal cavity, 3) degeneration of respiratory epithelium in larynx, pharynx and trachea, and 4) degeneration of dorsal skin were higher compared with Aldh2 +/+ mice, indicating that Aldh2 -/- mice are more acetaldehyde-sensitive than Aldh2 +/+ mice. This is the first example for studying pathological effects of Aldh2 deficiency using Aldh -/- mice exposed to a low level of acetaldehyde. PMID- 17127432 TI - Structure-proteasome-inhibitory activity relationships of dietary flavonoids in human cancer cells. AB - Diet high in vegetables and fruits has been associated with reduced cancer risk. However, the involved mechanisms are unknown. Previously, we reported that the dietary flavonoid apigenin could inhibit the proteasome activity and induce apoptosis in tumor cells. To further investigate the structure-proteasome inhibitory activity relationships, we chose and tested five dietary flavonoids, including luteolin, apigenin, chrysin, naringenin and eriodictyol. We found that the order of inhibitory potencies and apoptosis-inducing potencies of these five compounds in 20S purified proteasome and tumor cells was: (1) luteolin > apigenin > chrysin, and (2) apigenin >> naringenin, and luteolin >> eriodictyol. Therefore, flavonoids with hydroxylized B ring and/or unsaturated C ring are natural potent proteasome inhibitors and tumor cell apoptosis inducers. Furthermore, neither apigenin nor luteolin could inhibit the proteasome and induce apoptosis in non-transformed human natural killer cells. This finding may provide a molecular basis for the clinically observed cancer-preventive effects of fruits and vegetables. PMID- 17127433 TI - Electrochemical studies oxidation of ciprofloxacin at nano-SnO2/PVS modified electrode and its interaction with calf thymus DNA. AB - In pH 6.0 phosphate buffer solutions (PBS), a glassy carbon electrode (GCE) modifying by nano-tin oxide/polyvinyl sulfonic potassium (nano-SnO2/PVS) exhibited an enhanced effectiveness for the oxidation of ciprofloxacin (CFX), which compared with a bare GCE or a nano-SnO2 modified electrode. In addition we also investigated the electrooxidation mechanism of the fluoroquinolone antibiotics (utilizing ciprofloxacin, ofloxacin, sparfloxacin and lomefloxacin) at the modified electrode. Furthermore, gel electrophoresis coupled with electrochemistry and spectra techniques were used to study the interaction of CFX and calf thymus DNA (ctDNA). These acquired data showed that the binding mode of CFX and DNA was mainly an intercalation mechanism. PMID- 17127434 TI - Expression and function of utrophin associated protein complex in stretched endothelial cells: dissociation and activation of eNOS. AB - Several studies have emphasized the relevance of dystrophin-associated protein complex (DAPC) to maintain the vascular function. Previously we postulated the presence of an utrophin associated protein complex (UAPC) in endothelium from umbilical cord vessels. In the present work, we demonstrate that utrophin (UTR) indeed forms a complex, with beta-dystroglycan (DG), epsilon-sarcoglycan (SG), caveolin-1 (cav-1), and endothelial nitric oxide synthase (eNOS) in human umbilical vein endothelial cells (HUVEC) by co-immunoprecipitation analysis. Additionally, we observed an increment in the protein levels of epsilon-SG, beta DG, UTR and cav-1 after mechanical stretching. Interestingly, this stimulus also induced eNOS up-regulation, activation and release from the UAPC, and led to a significant increase in nitric oxide (NO) production. Finally, we propose that UAPC in HUVECs may play an important role in the regulation of vascular tone. PMID- 17127435 TI - Activated protein C in sepsis and beyond: update 2006. AB - Activated protein C (APC), a plasma serine protease, is best known for its ability to inhibit blood clot formation. APC acts as an anticoagulant by degrading coagulation cofactors Va and VIIIa, thereby attenuating the coagulation cascade. Over the past 15 years, impressive research advances have provided novel insights into the diverse biological activities of this molecule. APC is now viewed not only as an anticoagulant but also as a signaling molecule that provides a pivotal link between the pathways of coagulation, inflammation, apoptosis, and vascular permeability. The protective effect of APC supplementation in patients with severe sepsis likely reflects the ability of APC to modulate multiple pathways implicated in sepsis pathophysiology. This review attempts to summarize key studies that support the therapeutic potential of APC in conditions beyond sepsis such as stroke, ischemia-reperfusion injury, lung injury, asthma, pancreatitis, wound healing, and angiogenesis. A comprehensive PUBMED literature review up to May 2006 was conducted. PMID- 17127436 TI - A role for procarboxypepidase U (TAFI) in thrombosis. AB - The maintenance of the equilibrium between coagulation and fibrinolysis is crucial for normal haemostasis. In contrast, pathologic consequences of imbalance manifest tendencies of bleeding or thrombosis. Procarboxypeptidase U (proCPU, TAFI) is recognized as an important link between the coagulation system and fibrinolysis. Following activation by thrombin (IIa), carboxypeptidase U (CPU) exerts an antifibrinolytic effect by abolishing the cofactor function of partially degraded fibrin in plasminogen (Pg) activation. This review article focuses on the role of the proCPU/CPU system in the balance between fibrin deposition and removal. How a disturbed system can lead to a higher thrombotic tendency is discussed, while CPU inhibition as a new drug target for fibrinolytic therapy is extensively reviewed. PMID- 17127437 TI - Clinical applications of granulocyte-colony stimulating factor. AB - Granulocyte colony stimulating factor (G-CSF) is a naturally occurring potent neutrophil growth factor. Recombinant human G-CSF has been developed by pharmaceutical companies, and since the late 1980's, multiple clinical trials have explored its efficacy in a variety of medical conditions. These include various inherited and acquired neutropenia, as well as mobilization of hematopoietic stem cells and progenitors for transplantation. Interestingly, in several type of inherited neutropenia where no randomized controlled studies have ever been conducted, its chronic use is considered critical for survival and deemed a standard of care. Unfortunately, it the settings of cancer treatment related neutropenia and post hematopoietic stem cell transplantation, controversy still prevails whether universal usage of drug is cost effective despite innumerous randomized clinical trials. This review will focus on the clinical applications of G-CSF in the setting of inherited and acquired bone marrow failure, cancer treatment-related neutropenia and hematopoietic stem cell transplantation. PMID- 17127438 TI - Type-2 diabetic Lepr(db/db) mice show a defective microvascular phenotype under basal conditions and an impaired response to angiogenesis gene therapy in the setting of limb ischemia. AB - Diabetes mellitus is associated with macro- and micro-angiopathy, leading to increased risk of peripheral ischemia. In the present study, we have characterized the microvascular phenotype at the level of limb muscles and the spontaneous angiogenesis response to surgically-induced unilateral limb ischemia in a murine model of type-2 diabetes, the obese C57BL/KsOlaHsd-Lepr(db/db) mice (Lepr(db/db)), and in non-diabetic heterozygous Lepr(db/+). Wild type C57BL mice (WT) were used as controls. The basal microvascular phenotype was determined in mice aged 3 or 5 months, while the response to limb ischemia was studied only in 5-month old mice. Moreover, in 5-month old ischemic Lepr(db/db) and Lepr(db/+), we have tested the therapeutic potential of local angiogenesis gene therapy with human tissue kallikrein (hTK) or constitutively-activated Akt kinase (Myr-Akt). We found that in the muscles of 3- or 5-month old Lepr(db/db), apoptosis of endothelial cells was enhanced and the densities of capillary and arteriole were reduced. Arterioles of Lepr(db/db) showed hypertrophic remodelling and, occasionally, lumen occlusion. Following ischemia, Lepr(db/db) showed a defective reparative angiogenesis in ischemic muscle, delayed blood flow recovery, and worsened clinical outcome as compared with controls. Five-month old Lepr(db/+) displayed an increase in endothelial cell apoptosis under basal conditions, while capillary and arteriole densities were normal. Lepr(db/+) mounted a proper reparative angiogenesis response to limb ischemia and regained blood flow to the ischemic limb, regularly. Local gene therapy with hTK or Myr-Akt induced angiogenesis in ischemic muscles of Lepr(db/+) and Lepr(db/db). However, in the Lepr(db/db) neither gene therapy approach improved the blood flow recovery and the clinical outcome from ischemia. In contrast, either hTK or Myr-Akt gene transfer improved the post-ischemic recovery of Lepr(db/+). Type-2 diabetes has a negative impact on the basal microvascular phenotype and severely impairs post ischemic recovery of limb muscles. Gene therapy-induced stimulation of neovascularization might not suffice as a sole therapeutic strategy to combat type-2 diabetes-related vascular complications. In type-2 diabetic patients, therapeutic angiogenesis may need to be further optimized before being recommended for clinical applications. PMID- 17127439 TI - Alpha-tocopheryl phosphate is a novel apoptotic agent. AB - Alpha-tocopheryl succinate (TOS) is a well-known potent and selective apoptotic agent. This apoptotic activity has been ascribed to its detergent-like property which is also shared by the structurally related compound, alpha-tocopheryl phosphate (TOP). TOP meets the structural requirements that have been described for the apoptotic activity of TO esters, i.e. the combination of three structural, one functional, one signalling and one hydrophobic domain. In this study, we have investigated the effect of TOP on the osteosarcoma cell line MG-63 using TOS as a reference compound. As compared with TOS, TOP showed a higher proliferative and apoptosis inducing activity on the MG-63 cancer cell line. The cytotoxic effect of TOP and TOS seems to be due to the effect of the intact compounds, since only a minor conversion into alpha-tocopheryl (TO) could be detected. EPR experiments showed that TOS and TOP reduced membrane fluidity, whereas TO had no effect. In addition, induction of erythrocyte hemolysis by TOP depended on the pH. These results suggest that the detergent-like activity of these compounds might be involved in their biological effect. Due to the potent biological activities, TOP might be clinically useful. PMID- 17127440 TI - Mycoplasma genomics: tailoring the genome for minimal life requirements through reductive evolution. AB - Prokaryotic organisms of the genus Mycoplasma are characterized by their small body and genome size containing a 0.6-1.35 M bp genome. The genome is noted for its low G+C frequency ranging from 8-40 mol%. The Mycoplasma genus stems from the class Mollicutes (for soft skin), which lacks the cell walls and external motility appendages often present in other bacteria. To date, there are more than 100 known species of Mycoplasma. 34 species have been partially or completely sequenced. Widely known pathogenic species of Mycoplasma include: M. pneumoniae, causing pneumonia and other respiratory disorders, and M. genitalium, which are involved in pelvic inflammatory disease. Because of their small genome size, Mycoplasmas provide researchers a unique model of the minimal genomic requirements to maintain life. As the number of complete Mycoplasma genomes increase, these organisms become more established, thus laying the foundation for mapping evolutionary development. This manuscript provides an overview and update on Mycoplasma research, with particular focus on current genomics. PMID- 17127441 TI - Antioxidants decrease reperfusion induced arrhythmias in myocardial infarction with ST-elevation. AB - In myocardial infarctions with ST-segment elevation, ischemia followed by reperfusion (IR) leads to arrhythmia, myocardial stunning and endothelial dysfunction injury by reactive oxygen species (ROS). To determine the impact of ROS, we examined the effect of antioxidant vitamins on biochemical changes and arrhythmias induced by reperfusion before and after therapeutic thrombolysis (Actilyse). As compared with those receiving placebo, in individuals who received antioxidants, there was a significant decrease in premature ventricular beats (100% vs 38%), atrial fibrillation (44% vs 6%), ventricular tachycardia (31% vs 0%), first-degree atrial-ventricular block (44% vs 6%), plasma malondialdehyde at the first hour after initiation of thrombolysis (1.07 +/- 0.10 vs 0.53 +/- 0.10 nmols plasma malondialdehyde/mg protein) and circulating neutrophils after 24 hr after reperfusion. The antioxidant capacity of plasma was increased from 1.89 +/- 0.15 to 3.00 +/- 0.31 units/mg protein and paraoxonase-1 rose from 0.77 +/- 0.08 to 1.27 +/- 0.11 nmol/min/mg protein. These findings suggest that antioxidants might be useful as adjuvants in controlling reperfusion induced arrhythmias following therapeutic alteplase thrombolysis. PMID- 17127442 TI - Cytoplasmic binding partners of the platelet integrin alphaIIb beta3. AB - Platelets function physiologically in mediating hemostasis, but are also associated with many pathological conditions, such as thrombosis, which can lead to myocardial infarction and/or stroke. Therefore, the study of platelet regulation and signaling has been of great interest and is necessary for generating effective anti-platelet therapeutics. One platelet signaling molecule of particular interest is the integrin alphaIIb beta3, which binds Fg and mediates platelet cross-linking. The integrin itself as well as cytoplasmic proteins that interact with alphaIIb beta3 have become potential targets for anti platelet therapies. One such protein that has been shown to directly regulate alphaIIb beta3 function is calcium- and integrin-binding protein 1 (CIB1). CIB1 has been implicated in alphaIIb beta3 activation and outside-in signaling through the integrin. By increasing our understanding of CIB1 and other proteins that like it, associate with integrin alphaIIb beta3, and the signaling events that result from those interactions, we may bring ourselves closer to more effective therapies. In the present work, we explore known cytoplasmic binding partners of the integrin alphaIIb beta3 and their effect on alphaIIb beta3, focusing on CIB1. PMID- 17127443 TI - Megakaryopoiesis: transcriptional insights into megakaryocyte maturation. AB - Platelets are small anucleate cells that travel near the vessel wall during laminar flow. In response to vascular injury, platelets undergo alterations in morphology which allow them to aggregate and cover the injured site. Platelets are produced by megakaryocytes in a process that involves the formation of platelet precursors called proplatelets and subsequent release of these proplatelets into the circulation. By forming a demarcation membrane system within the cytosol, megakaryocytes contain a membrane reservoir which allows for the production of thousands of platelets per mature megakaryocyte. Interestingly, the above process known as megakaryopoiesis is not yet fully understood. However, several groups have contributed evidence to unveil the role of thrombopoietin (TPO), the principal regulator of megakaryopoiesis in vivo. TPO is necessary for megakaryocyte maturation in that TPO deficient mice display greatly reduced megakaryocyte production as well as reduced numbers of mature megakaryocytes. Several transcription factors have also been implicated in megakaryopoiesis including, GATA-1, friend of GATA-1 (FOG-1), nuclear factor-erythroid 2 (NF-E2), and Fli-1. In fact, interactions among some of the transcription factors have been reported to produce synergistic effects. GATA-1 and Fli-1 interactions result in heightened GPIX and GPIb (2 components of von Willebrand Factor (vWF) receptor) expression, while GATA-1, RUNX1 and core-binding factor beta interactions result in improved alphaIIb promoter activity. Mutations in the vWF complex and alphaIIb beta3 have been linked to disorders such as Bernard-Soulier syndrome and Glazmann thrombasthenia respectively. Therefore, a more comprehensive understanding of the transcriptional control of megakaryopoiesis may lead to more effective treatments of platelet-related disorders. PMID- 17127444 TI - Recent advances in high-throughput molecular marker identification for superficial and invasive bladder cancers. AB - Bladder cancer is the fifth most common neoplasm in industrialized countries. Due to frequent recurrences of the superficial form of this disease, bladder cancer ranks as one of the most common cancers. Despite the description of a large number of tumor markers for bladder cancers, none have individually contributed to the management of the disease. However, the development of high-throughput techniques for simultaneous assessment of a large number of markers has allowed classification of tumors into clinically relevant molecular subgroups beyond those possible by pathological classification. Here, we review the recent advances in high-throughput molecular marker identification for superficial and invasive bladder cancers. PMID- 17127445 TI - Apo2L/TRAIL and immune regulation. AB - Apo2L/TRAIL is a member of the TNF family, with its receptors DR4 and DR5 containing a death domain. Multiple tumors are sensitive to Apo2L/TRAIL-induced apoptosis, while normal cells are not, so it constitutes a promising new antitumoral therapy. In this review we deal rather with the physiological role of Apo2L/TRAIL, which, in one hand, is clearly related with immune antitumoral surveillance. However, a role of Apo2L/TRAIL as a fine-tuning regulator of the immune system, especially in the regulation of CD8+ T cell activation and memory, has been also demonstrated. In fact, Apo2L/TRAIL can be considered as an additional mechanism needed to prevent the development of autoimmune disease. Indeed, recent developments indicate that Apo2L/TRAIL can be also useful as a treatment against certain chronic autoimmune diseases. PMID- 17127446 TI - Polyalanine expansion mutations in the X-linked hypopituitarism gene SOX3 result in aggresome formation and impaired transactivation. AB - Polyalanine expansion mutations have been identified in eight transcription factors that are associated with a range of congenital disorders. While some of these mutant proteins have been shown to generate cellular aggregates in heterologous cell lines, little is known about the mechanism by which these aggregates cause disease. Here we examine the aggregation and functional properties of the two known polyalanine expansion mutations associated with X linked Hypopituitarism (XH), SOX3(22Ala) and SOX3(26Ala), which contain an additional seven and eleven alanine residues, respectively. SOX3(22Ala) and SOX3(26Ala) proteins form cytoplasmic aggregates and nuclear inclusions in transiently transfected COS-7 and CHO K1 cells, and in transfected explant cultures of chick neural epithelium. SOX3(26Ala) exhibits a more potent aggregation phenotype, resulting in significantly more cells with dispersed cytoplasmic and large perinuclear aggregates. SOX3(22Ala) and SOX3(26Ala) protein aggregates exhibit the key properties of aggresomes including vimentin redistribution, colocalisation with the Microtubule Organising Centre and sensitivity to microtubule disruption. This is the first time that aggresomes have been implicated in the aetiology of a polyalanine expansion disorder, suggesting that XH and protein conformation disorders may become manifest through similar pathological mechanisms. Further, we show that mutant SOX3 proteins have impaired transcriptional activity and reduced capacity to inhibit beta catenin/TCF-mediated transcription. These data suggest that deregulation of SOX3 target genes and inappropriate canonical Wnt signaling in central nervous system (CNS) progenitors may also contribute to dysfunction of the hypothalamic pituitary axis in XH patients. PMID- 17127447 TI - Tracing complement-retroviral interactions from mucosal surfaces to the lymphatic tissue. AB - During (nearly) all steps in retroviral pathogenesis, viruses are confronted with complement and complement receptor (CR)-positive cells. As all of the retroviruses tested so far activate the complement system, members of this virus family have adapted different protection mechanisms to keep complement activation under the threshold necessary to avoid complement-mediated lysis. As a consequence of complement activation, retroviruses are covered with complement proteins and thus provide additional ligands to interact with CR-expressing cells. This review discusses the complex complement-retroviral interactions and follows the fate of the virus on its way to the lymphatic tissue. PMID- 17127448 TI - A single vaccination with attenuated SIVmac 239 via the tonsillar route confers partial protection against challenge with SIVmac 251 at a distant mucosal site, the rectum. AB - Elucidating the mechanisms that protect monkeys previously immunized with attenuated SIV (SIVDeltanef) against challenge infection with pathogenic virus may reveal new strategies for the development of an effective HIV vaccine. Here we show that a single atraumatic application of SIVDeltanef to the tonsils of four rhesus macaques conferred protection against SIVmac251 applied intrarectally 26 weeks later. While this protection was not complete, i.e., challenge virus could be isolated from all immunized animals, it was reflected by significantly lower viral loads in the blood (weeks 2-16 after challenge, p < 0.01) and considerably lower loads in lymphoid organs, and more stable peripheral CD4 counts in a proportion of the immunized animals as compared to four non immunized, SIVmac251-infected control monkeys. SIV-specific humoral as well as systemic and mucosal T cell responses were detected in the immunized animals, but there was no correlation between their magnitude of expression and the level of protection. Analyses of leukocyte subsets in these animals at necropsy (24 weeks after challenge) did not reveal a significantly enhanced proportion of gamma/delta T cells in the tissues of protected monkeys. Therefore, tonsillar application of attenuated SIV induces protection in some animals against a superinfection with wild-type SIV distant at a distant mucosal site. PMID- 17127449 TI - Float on: lipid rafts in the lifecycle of HIV. AB - All steps in the HIV lifecycle--entry, assembly, budding, induction of signal transduction and subsequent cell activation--are complex multifactorial mechanisms where cholesterol and sphingolipids (glycosphingolipids--GSLs and sphingomyelins in mammalian cells) are closely involved. Here we will review the contribution of these heterogeneous membrane lipid microdomains, referred to as lipid rafts, DIGs (detergent-insoluble glycolipid-enriched complexes), DRMs (detergent-resistant membranes), GEMs (glycolipid-enriched membranes), caveolae, TIMs (Triton-insoluble membranes) for interactions of HIV with the host cell. The accurate terminology was discussed elsewhere, and to simplify matters we will use rafts or lipid rafts throughout the review. PMID- 17127450 TI - B cell receptor editing in tolerance and autoimmunity. AB - The random assembly of immunoglobulin (Ig) genes often creates a B cell receptor that is self-reactive, and such cells are subjected to negative selection. A primary mechanism to extinguish this self-reactivity is receptor editing, which allows continued recombination of Ig genes and replacement of the self-reactive receptor with a new innocuous receptor. Recent data now suggest that receptor editing may also promote autoimmunity in an autoimmune context. This mechanism has also been implicated in the process of B cell positive selection and maturation. Here we discuss the contribution of receptor editing in B lymphopoiesis and its importance for B cell tolerance and autoimmunity. PMID- 17127451 TI - Mitochondrial contribution to the molecular mechanism of heart acclimatization to chronic hypoxia: role of nitric oxide. AB - A remarkable number of adaptive responses; including changes in the cardiovascular, respiratory and hematologic systems; takes place during acclimatization to natural or simulated high altitude. This adaptation to chronic hypoxia confers the heart an improved tolerance to all major deleterious consequences of acute O2 deprivation, not only reducing infarct size but also alleviating post-ischemic contractile dysfunction and ventricular arrhythmias. There is growing evidence about the involvement of mitochondria and NO in the establishment of cardioprotection. This review focuses on evidence about the putative role of different effectors of heart acclimatization to chronic hypoxia. Along with classical parameters, we consider NO, specially that generated by mtNOS, mitochondrial respiratory chain, mitoK(ATP) channels, reactive oxygen species and control of gene expression by HIF-1. PMID- 17127452 TI - Changes in HIV-specific antibody responses and neutralization titers in patients under ART. AB - In this study, we tested for antibody reactivities against gp120 and gp41-derived peptides, recombinant gp160, gp41 and tat in HIV-positive sera under antiretroviral therapy (ART) and determined their neutralization capacity. As a baseline, sera from patients in stage A, B and C of the disease, long term non progressors (LNPs) and HIV-negative individuals were included. Compared to LNPs or sera from patients in group A, the reactivity of sera in stage B or C against gp120-derived peptides was reduced parallel to disease progression. Reactivity of these samples was compared with sera of patients under ART. Parallel to the decrease of viral load, the reactivity against gp120 and gp41-derived epitopes, recombinant gp160 and gp41 or the native gp120/41 complex was significantly reduced. Antibody-mediated neutralization of HIV-1 was detectable prior to ART but revealed substantial decreases coupled with progression of therapy. Responses to recombinant tat dropped after three months of therapy, increased however at later time points to initial levels. These data indicate that in parallel to the decrease in viral load and antibodies against gp120, the neutralization capacity of sera under ART is reduced, and can not be compensated by an increase in tat specific antibodies. PMID- 17127453 TI - Role of B cells in Sjogren's syndrome--from benign lymphoproliferation to overt malignancy. AB - The classical view of B cell biology is that these cells respond to foreign and self antigens and in this way promote protection, primarily by production of antibodies. However, recent studies suggest that B cells have diverse functions within the immune system other than antibody production, which could contribute to autoimmunity. This involves organization of lymphoid tissue, regulation of dendritic cells, antigen presentation, activation of T cells and production of cytokines. Both abnormalities in the distribution of B cell subsets, and recent discovery of clinical benefit after B cell depletion highlight the pivotal role of B cells in autoimmunity. This change in view of the role of B cells will be exemplified in one autoimmune disease namely Sjogren's syndrome. PMID- 17127454 TI - Atherosclerosis, antiphospholipid syndrome, and antiphospholipid antibodies. AB - In antiphospholipid syndrome (APS) patients, some antiphospholipid antibodies (APA) are directed against negatively-charged phospholipids, while other APA are specific for phospholipid-proteins such as beta2-glycoprotein I (beta2GPI). Increased levels of oxidized low density lipoproteins (oxLDL) are present in atherosclerosis patients and these patients develop anti-oxLDL antibodies. Several studies have reported cross-reactivities between APA and anti-oxLDL antibodies, and some authors have suggested that most APA are specific for oxidized forms of phospholipids. In contrast, other studies report that APA react with both reduced and oxidized cardiolipin. In this context, we have re-examined the literature surrounding antibodies found in atherosclerosis and the APS. We have also included results from a panel of anti-phospholipid monoclonal antibodies from W/B mice, an APS model, which indicates that these antibodies do not display any preference for oxidized epitopes on lipid molecules. PMID- 17127455 TI - Invasion of the killer B's in type 1 diabetes. AB - Type 1 Diabetes (T1D) is an autoimmune disease requiring contributions from effectors in both CD4+ and CD8+ T cell compartments in order to destroy insulin producing pancreatic beta cells. Autoantibodies specific for islet cell proteins are also often generated during the prodromal stages of T1D development. While providing excellent prognostic markers of future disease risk, it has generally been believed that the induction of autoantibody secretion by B cells was a secondary consequence of the ongoing autoreactive T cell response. However, studies in the NOD mouse model of disease have demonstrated that B cells play a key function during T1D development by serving as a subpopulation of antigen presenting cell (APC) which can most efficiently support the expansion of diabetogenic CD4+ T cells. Furthermore, studies utilizing this model have indicated that autoantibodies may play an important role in initiating an early phase of pancreatic beta cell destruction ultimately leading to overt T1D. This review will focus on the under appreciated role B cells play in T1D development not only in NOD mice, but also potentially in humans. PMID- 17127456 TI - Anti-CD20 therapy and autoimmune disease: therapeutic opportunities and evolving insights. AB - Based on the successful clinical experience with the anti-CD20 antibody, rituximab, for the treatment of B-cell non-Hodgkins lymphoma, there is a rapidly growing literature on the treatment of patients with autoimmune diseases with this therapeutic agent. However, the pathogenetic mechanisms responsible for these diseases may differ greatly from those in B cell malignancies. Herein, I provide an overview on recently published clinical experience, and discuss immunobiologic perspectives that are most relevant to understanding the special opportunities and challenges posed by these diseases. Of special importance, there is emerging evidence that the same inherited genetic variations and acquired immunodefects that underlie autoimmune disease pathogenesis may in some patients also interfere with the efficacy of anti-CD20 antibody-based therapy. PMID- 17127457 TI - Three pathogenic determinants in immune nephritis--anti-glomerular antibody specificity, innate triggers and host genetics. AB - The prevailing notion is that lupus nephritis is mediated by autoantibodies, particularly those that bind to DNA and/or glomeruli. However it has become apparent that the development of immune-mediated renal disease is contingent upon additional factors including innate stimuli and host genetics. The purpose of this review is to evaluate our current understanding of three factors that can potentially influence immune-mediated renal disease: (1) Anti-glomerular/DNA antibodies (Abs), (2) Innate triggers, including Toll-Like Receptor (TLR) stimulation, and (3) the genetic makeup of the host. PMID- 17127458 TI - Direct AAV-mediated gene delivery to the temporomandibular joint. AB - Successful intra-temporomandibular joint gene transfer is a prerequisite for gene therapy to induce mandibular condylar growth. This study was carried out to investigate the expression of recombinant adeno-associated virus (rAAV) based vector-mediated enhanced green fluorescence protein (eGFP) in the mandibular condyle. The transduction efficiencies for primary chondrocytes using different hybrids of rAAV vectors were identified by fluorescence microscopy and FACS. Sprague-Dawley rats were injected with either rAAV(2/2)-eGFP construct or saline into both mandibular condyles. The spatial and temporal transgene expression was detected by in situ hybridization, RT-PCR and real-time PCR. Our result showed that rAAVs were capable of infecting rat chondrocytes. rAAV(2/2) was the best serotype to infect chondrocytes in vitro. By in situ hybridization, eGFP expression was clearly detected in the deeper layer of mandibular condyle as early as 7 days after injection. By real-time PCR, the transgene expression reached a peak at 21 days. This study was the first report to explore that rAAV mediated genes could be transferred to mandibular condyle in vivo. Our results strongly suggest that rAAV(2/2) mediated gene delivery is a promising approach to deliver candidate genes for future regulating mandibular condylar growth. PMID- 17127459 TI - Gene expression profiling of early involuting mammary gland reveals novel genes potentially relevant to human breast cancer. AB - Mammary gland involution represents one of the most dramatic examples of programmed cell death/apoptosis and tissue regression during development, yet large gaps still exist in the understanding of the mechanisms involved, and the key factors that trigger involution, are not yet identified. With the focus on identifying "novel" genes associated with mammary gland regression, we used microarray analysis to examine differentially expressed genes during early mammary gland involution in the mouse. We then examined the relevance of candidate genes to human tumorigenesis and identified a number of genes not previously implicated or not well characterized in human breast cancer. The expression levels of these genes in human breast cancer were confirmed in breast cancer cell lines and breast tumor tissues. This pilot study demonstrates proof of principle that through the analysis of gene expression during mammary gland involution, it may be possible to identify "novel" genes relevant human breast cancer. PMID- 17127460 TI - Tumor suppressor loci in bladder cancer. AB - Large numbers of genetic and epigenetic alterations have been identified in bladder cancer in recent years. Many of these affect the function of tumour suppressor genes (TSGs), leading to partial or complete loss of protein expression or function with varied phenotypic consequences. Some of the genes implicated such as TP53 and RB1 are major players in many other tumour types. Others, particularly some on chromosome 9, show bladder-specific involvement. Other TSGs of relevance to bladder tumour development are predicted by the finding of common physical deletion or LOH in specific regions of the genome. This review summarises the approaches that have been used to identify bladder tumour suppressor genes, the current state of knowledge of the genes involved in this disease, their relationship with specific clinical features and some possible therapeutic applications. PMID- 17127461 TI - Proliferative status of tumor stem cells may be correlated with malignancy grade of human astrocytomas. AB - Tumor stem cells are implicated in tumor initiation and maintenance. Recent studies have shown that a subpopulation of cells isolated from brain tumors can form neurospheres in vitro, and have multiple characteristic properties observed in neural stem cells. In vivo implantation of these cells can induce tumors that phenocopy original tumors, suggesting that tumor stem cells are involved in brain carcinogenesis. We found that a population of cells in human glioblastoma multiforme expressed multiple protein markers of neural stem cells including nestin, TUC-4, doublecortin and beta III-tubulin. In contrast, these markers were not expressed in human capillary hemangioblastoma or meningioma. Double immunolabeling showed that a portion of doublecortin-, beta III-tubulin-, TUC-4- and nestin-positive cells express Ki67 antigen, a cell proliferation marker. To investigate further whether these properties of tumor stem cells are correlated with their biological behavior, immunohistochemistry was performed on brain sections from astrocytomas of different grades using antibodies against neural stem cell markers. The number of cells expressing Ki67 antigen and neural stem cell markers was increased in relation to worsening histological grade of astrocytomas, indicating that the capacity for tumor stem cell proliferation may be clinically relevant. Thus, tumor stem cells in astrocytomas may be involved in carcinogenesis. PMID- 17127462 TI - Combination of 4-HPR and oral contraceptive in monkey model of chemoprevention of ovarian cancer. AB - 4-(N-hydroxyphenyl) retinamide (4-HPR) and the oral contraceptives (OCP) are currently being used alone, and in combination, for the prevention of ovarian cancer. However, the mechanism of their effects has not been studied. Non-human primate models are ideal for studying the role of these and other drugs for cancer chemoprevention because of the genetic similarity between primates and humans in respect to hormone regulation and menstrual cycle. 4-HPR and OCP were administered to sixteen female adult Macacca mulatta (Rhesus macaques) for three months alone and in combination. Laparotomy was performed before and after treatment, and ovarian biopsies were obtained to evaluate the expression of retinoid and hormone receptors, and apoptosis. ER alpha was undetectable, but ER beta, PR, RXR alpha, and RXR gamma were constitutively expressed in the ovaries. 4-HPR induced RXR alpha and RXR gamma expression at a low level and, OCP induced expression of ER beta. However, the combination of 4-HPR with OCP had a larger effect on expression of retinoid receptors. Apoptosis was detected in the 4-HPR group (equivalent dose: 200 mg/day). PMID- 17127463 TI - Ets-1 participates in and facilitates developmental expression of hypoxia-induced mitogenic factor in mouse lung. AB - Hypoxia-induced mitogenic factor (HIMF) possesses mitogenic, vasoconstrictive, angiogenic, and antiapoptotic effects. While HIMF is known to be expressed in developmental mouse lung, its gene expression regulation during this period is completely unknown. Genomic sequencing of HIMF gene has shown that there is an Ets-1 binding site in its 5'-promoter-region. To test the hypothesis that Ets-1 protein expressed in developing mouse lung may participate in the process of HIMF gene expression regulation via direct involvement or facilitation, we characterized the proximal promoter of HIMF gene (409-bp long fragment that includes the -347-bp promoter region from transcription start site), and investigated HIMF and Ets-1 expression with western blot and immunohistochemistry, electrophoretic mobility shift assay (EMSA) for Ets-1, HIMF promoter luciferase reporter gene assays, and chromatin-immunoprecipitation (CHIP). Western blots revealed that both Ets-1 and HIMF proteins were developmentally expressed in the lung. Immunohistochemical localization revealed Ets-1 signals in the nucleus of HIMF-expressing airway epithelial cells and alveolar type II cells, whereas HIMF was localized in cytoplasm. Presence of Ets 1 protein within E16, E20, and adult lung nuclear extract was demonstrated by EMSA. Co-transfection of Ets-1 expression plasmid with HIMF promoter construct increased luciferase activity in NIH3T3 cells, but mutation or deletion of Ets-1 site eliminated HIMF promoter luciferase activity. CHIP with anti-Ets-1 antibody revealed Ets-1 binding to HIMF promoter region in E20 and P1 but not in E15 lung. We conclude that Ets-1 participates in the process of HIMF gene expression and Ets-1-mediated HIMF expression may play an important role in lung development and maturation. PMID- 17127464 TI - Regulation of vascular endothelial growth factor, VEGF, gene promoter by the tumor suppressor, WT1. AB - Understanding angiogenesis and growth control is central for elucidating prostate tumorigenesis. However, the mechanisms of activation of the angiogenic gene, vascular endothelial growth factor (VEGF) are complex and its regulation in prostate cancer is not well understood. In previous studies, VEGF expression levels were correlated with altered levels of the zinc finger transcription factor, WT1. Since the VEGF promoter has several potential WT1 binding sites and WT1 regulates many growth control genes, here we assessed whether WT1 might also regulate VEGF transcription. Using transfection and DNA binding assays, functional WT1 binding sites were localized within the proximal VEGF promoter. Transfection of the DDS-WT1 (R394W) zinc finger mutant had no significant effect on VEGF-luciferase reporter activity, suggesting that an intact zinc finger DNA binding domain was required. Interestingly, WT1-mediated regulation of VEGF reporter constructs varied in different cell types. In androgen-responsive, LNCaP prostate cancer cells, hormone treatment enhanced WT1-mediated activation of the VEGF promoter constructs. Overall, these results suggest that WT1 transcriptionally regulates VEGF through interaction of its zinc finger DNA binding domain with the proximal GC-rich VEGF promoter. These findings may shed light on the role of WT1 in angiogenesis and prostate cancer progression. PMID- 17127465 TI - Blood glucose lowering effects of brown rice in normal and diabetic subjects. AB - Carbohydrate foods, which produce low glycemic responses, have been shown to be beneficial in the dietary management of chronic diseases such as diabetes and hyperlipidemia. This study determined the starch digestion rate in vitro and, in a randomised crossover design, the postprandial blood glucose response of 10 healthy and nine type 2 diabetic volunteers to brown rice compared to milled rice from the same batch and variety. The total sugar released in vitro was 23.7% lower in brown rice than in milled rice. In healthy volunteers, the glycemic area and glycemic index were, respectively, 19.8% and 12.1% lower (p < 0.05) in brown rice than milled rice, while in diabetics, the respective values were 35.2% and 35.6% lower. The effect was partly due to the higher amounts of phytic acid, polyphenols, dietary fiber and oil in brown compared to milled rice and the difference in some physicochemical properties of the rice samples such as minimum cooking time and degree of gelatinisation. In conclusion, brown rice is a more health beneficial food for diabetics and hyperglycemic individuals than milled rice. PMID- 17127466 TI - Dietary habits and related psychological and social factors: influence on the body weight of elementary school children. AB - This article studies the alimentary habits of children in relation to their body mass index (BMI) and food preferences, and to underline the influences of cultural and social aspects. The investigation tested 198 children (90 male and 108 female) from Cento (Ferrara), using food frequency questionnaires that were completed by the children. Subjects were subdivided into three groups based on their BMI--normal weight, overweight and underweight--taking into consideration their food frequency and preferences. The correlations between the BMI of children, mother's geographic origin and cultural level revealed differences between boys and girls. The variability of results does not allow precise correlations to be made between food frequency and children's weight, psychological and social aspects. Some incorrect dietary habits were revealed in the overweight and underweight groups. These could be controlled for and corrected by an alimentary education programme. PMID- 17127467 TI - Comparative activity of antioxidants from wheat sprouts, Morinda citrifolia, fermented papaya and white tea. AB - Hydroalcoholic extracts from wheat sprouts, white tea, Morinda citrifolia and fermented papaya were analysed to determine their reducing power and antioxidant activity. The results show that the micromoles of potassium ferricyanide reduced by a quantity of extract corresponding to 1 g of the various dehydrated starting tissues are: 12.91+/-0.83 (wheat sprouts), 10.66+/-1.22 (M. citrifolia), 17.06+/ 1.24 (white tea), and 1.05+/-0.09 (fermented papaya). In addition the results show a strong oxygen superoxide scavenging activity in the extracts from white tea, M. citrifolia and wheat sprouts. The activity of the fermented papaya extract is the lowest. The thin-layer chromatography and UV spectrophotometry of the extracts show in each source a mixture of antioxidant compounds probably belonging to the families of reducing glycosides and polyphenols. The chromatographic pattern of the antioxidant compounds and the UV spectrum are quite different in the various sources. PMID- 17127468 TI - Structural modulation of calmodulin and calmodulin-dependent protein kinase II by pea protein hydrolysates. AB - The effects of two fractions of pea protein hydrolysate with high levels of positively charged amino acids on the structural conformations of calmodulin (CaM) and CaM-dependent protein kinase II (CaMKII) were determined using fluorescence and circular dichroism methods. In the presence of Ca2 + , addition of the protein hydrolysates to CaM and CaM/CaMKII complex led to increased exposure of aromatic groups as measured by intrinsic and extrinsic fluorescence spectroscopy. Near-UV circular dichroism data revealed an increase in the tertiary structure of CaM in the presence of pea protein hydrolysates. Effect of the protein hydrolysates on the CaM structure was greater with the fraction that contained higher contents of arginine and lysine when compared with the fraction with lower levels of these two amino acids. We concluded that the presence of the pea protein hydrolysates led to rearrangement of the native protein structure and exposure of buried hydrophobic groups of CaM and/or CaMKII. PMID- 17127469 TI - Calcium, iron and zinc uptakes by Caco-2 cells from white beans and effect of cooking. AB - White beans (Phaseolus vulgaris L.) have an interesting content of essential elements, calcium, iron and zinc, but they content also phytates, oxalates, proteins, polyyphenols and complex polysaccharides that are known to interact with minerals and to affect their bioavailability. The bioavailability of calcium, iron and zinc from raw and cooked white beans was estimated using their uptake by Caco-2 cells as the criteria. Previously, the mineral fraction (soluble or dialysable) to be added to the Caco-2 cell monolayer was selected. The results obtained show that cooking increases the Caco-2 cells' uptake percentages (calcium, 18.8 versus 3.6; iron, 33.7 versus 1.7; and zinc, 17.2 versus 2.1) and improved the value of beans as a dietetic source of minerals. PMID- 17127470 TI - Entomophagy among the Luo of Kenya: a potential mineral source? AB - Primary objective To determine the iron, zinc, and calcium content in different insects commonly eaten among the Luo of Kenya. Research design A cross-sectional design was chosen for the study in order to determine the insects eaten and their mineral content during a specific season.Methods and procedures Five different insect species were identified and collected with the help of local informants in the Nyang'oma sublocation of the Bondo district in western Kenya, and were analysed for iron, zinc and calcium contents. Main outcomes and results The iron content ranged from 18 to 1562 mg/100 g dry matter, the zinc content from 8 to 25 mg/100 g, and the calcium content from 33 to 341 mg/100 g in five different insects, onyoso mammon (ant), oyala (termite), ogawo (termite), agaor (termite), onjiri mammon (cricket). Conclusions Insect eating could prove to be a valuable measure to combat, especially, iron and zinc deficiency in developing countries. PMID- 17127471 TI - Effect of whey protein to modulate immune response in children with atopic asthma. AB - BACKGROUND: Levels of glutathione (GSH) in antigen-presenting cells promote a T helper type 2 (Th2) cytokine response in mice. We have previously demonstrated that we can increase intracellular GSH levels in healthy young adults using a whey-based oral supplement (HMS90). We hypothesized that such supplementation in children with atopic asthma, a Th2 cytokine disease, would improve lung function and decrease atopy. METHODS: Eleven children (six females, five males; mean+/ standard deviation age, 12.6+/-3.6 years; baseline forced expired volume in 1 sec (FEV1), 82.4+/-15.4%predicted), underwent spirometry, methacholine provocation testing, and blood analysis for serum IgE and lymphocyte GSH before and after 1 month of supplementation (10 g twice daily). RESULTS: Initially the IgE was 1689+/-1596 microg/l (normal range 0.90), and also linked (r2>0.80) to antioxidants (alpha tocopherol, glutathione and catalase). A further 10 high-resonance gas chromatography nitrogen phosphorus detector flavour components were correlated (r2>0.85) with C18:3(n-3) content. Chicken character was correlated with C18:3(n 3), and C18:3(n-6) inversely with oily, off-flavour and lipid oxidation. Sweet, fruity and oily aromas were linked in PLS1 with 13 specific fatty acids (r2>0.6), and bland taste with total summed (six) fatty acid fractions (r2>0.81). Specific antioxidants were correlated with sweet, fruity and chicken aromas, and alpha tocopherol inversely with lipid oxidation. PLS2 confirmed relationships between fatty acid composition, antioxidants and the subsets of 32 and 10 flavour components. Clear relationships were thus observed between lipid and antioxidant compositions and flavour in chicken breast meat. PMID- 17127475 TI - Effect of thermal processing on retinol levels of free-range and caged hen eggs. AB - Purpose Eggs are a food item of high nutritional value, a source of vitamin A and readily accessible to the general population. Methods This paper analysed the effect of cooking on the retinol levels of free-range and caged hen eggs, using high performance liquid chromatography (HPLC). The retinol levels of hen and quail eggs were also compared. Results The raw egg yolk retinol concentrations of free-range and caged hen eggs were 476.53+/-39.44 and 474.93+/-41.10 microg/100 g and cooked egg yolk concentrations were 393.53+/-24.74 and 379.01+/-30.78 microg/100 g, respectively; quail egg concentration was 636.56+/-32.71 microg retinol/100 g. No significant difference was found between the retinol of free range and caged hen egg yolks; however, cooking diminished retinol levels, causing a loss of 17 and 20% in the free-range and caged hen egg yolks, respectively. Quail egg retinol concentration was significantly higher than that of the hens. Conclusion The retinol found in 100 g of hen and quail egg yolks could supply around 42 and 70.7% of the vitamin A requirements of an adult man, and is accordingly considered an excellent source of this vitamin. PMID- 17127476 TI - Can pure fruit and vegetable juices protect against cancer and cardiovascular disease too? A review of the evidence. AB - While it is widely accepted that fruit and vegetables (F&V) lower the risk of cancer, and cardiovascular disease (CVD), the role of pure fruit and vegetable (PFV) juices is often downplayed. This review poses two questions: Are the protective benefits of F&V dependent upon constituents lacking in PFV juices (e.g. fibre)? Do PFV juices impact on disease risk when considered separately from F&V? Studies comparing the effects of fibre and antioxidants were reviewed, yielding the finding that the impact of F&V may relate more strongly to antioxidants, than to fibre. For the second question, high-quality published studies that considered PFV juices were reviewed. The impact of PFV juices on cancer risk was weakly positive, although a lack of human data and contradictory findings hampered conclusions. For CVD, there was convincing evidence from epidemiological and clinical studies that PFV juices reduced risk via a number of probable mechanisms. It was concluded that the view that PFV juices are nutritionally inferior to F&V, in relation to chronic disease risk reduction, is unjustified. PMID- 17127477 TI - Use of low-glycaemic index bread to reduce 24-h blood glucose: implications for dietary advice to non-diabetic and diabetic subjects. AB - The present study investigated the effect of a simple dietary change on 24-h blood glucose. In a randomized cross-over design, 10 healthy subjects were prescribed a low-glycaemic-index (LGI) diet and a high-glycaemic-index (HGI) diet. The diets were identical with the exception of the type of bread consumed (LGI or HGI). Glucose concentrations over 24 h were measured using a continuous glucose monitoring system. The LGI diet resulted in a lower mean glucose response compared with the HGI diet over 24 h (P=0.135), during the day (P=0.171) and at night (P=0.100). Similarly, the 24-h, daytime and overnight incremental area under the curve for glucose following the LGI diet was consistently lower than following the HGI diet (P=0.093, P=0.132 and P=0.061, respectively). The results demonstrate how a very simple dietary change can favourably alter overall blood glucose concentrations. Such small modifications to the diet, if adopted in the long term, could improve glucose control and consequently reduce the risk of chronic disease in both diabetic and non-diabetic individuals. PMID- 17127479 TI - Thrombelastography: current clinical applications and its potential role in interventional cardiology. AB - Thrombelastography is a bedside blood test used to assess patients' haemostatic status. It has a well-established role in hepatobiliary and cardiac surgery and is also used in obstetrics and trauma medicine to assess coagulation and identify the causes of post-operative bleeding. It is not routinely used in the diagnosis or treatment of thrombosis although recently it has been shown to predict thrombotic events post-operatively and after percutaneous intervention (PCI). In cardiovascular medicine the importance of the platelet in the pathophysiology of vascular events is increasingly apparent. As a result antiplatelet therapy is a cornerstone of the treatment for coronary disease, particularly in the setting of acute coronary syndromes. The increasing utilization of stents, particularly drug eluting devices, in PCI has also necessitated widespread use of antiplatelet agents to minimize the risk of stent thrombosis. A quick, accurate and reliable test to measure the effect of platelet inhibition by antiplatelet agents on clotting in an individual patient would be of profound clinical value. The results from such a test could provide prognostic information, allow treatment with antiplatelet agents to be tailored to the individual and identify resistance to one or more of these agents. Optimization and tailoring of anti-platelet therapy in patients with cardiovascular disease, particularly those undergoing PCI, using such a test may reduce morbidity and mortality from thrombotic and haemorrhagic complications. Current methods of assessing platelet activity measure platelet count and function in isolation. Optical aggregation is the most widely used method for assessing platelet function but it is relatively time consuming, measures platelet function in isolation rather than in the context of clot formation and is not a bedside test. By contrast the modified thrombelastograph platelet mapping kit marketed by Haemoscope can be used to assess the effects of antiplatelet agents on ex vivo blood clotting, thus giving a measurement more relevant to in vivo responses. This represents a potentially powerful tool to assess response of individual patients to antiplatelet therapy, particularly in the context of PCI. PMID- 17127480 TI - Gray Platelet Syndrome in a Somalian family. AB - The Gray Platelet Syndrome (GPS) is a rare inherited, hypogranular platelet disorder characterized by virtual absence of alpha granules in bone marrow megakaryocytes and circulating plates. Usually only one member of a family is affected, but families with two or more affected individuals have been reported from France, Australia, and Mexico, and, recently, the United States. The current study has evaluated the first family from East Africa with two affected members, a brother and sister. Neither child has had significant bleeding problems. Electron microscopic studies revealed almost complete absence of alpha granules from their platelets. Instead their platelets were filled with vacuoles similar in size to the missing granules. Dense bodies were normal in number in the girl's platelets, but markedly increased in her brother's cells. Tannic acid staining revealed that very few of the vacuoles were connected to channels of the open canalicular system. As a result, contents of the residual alpha granule vacuoles must leak out of the organelles and diffuse through megakaryocyte and platelet cytoplasm to the outside. The route of escape may differ from other hypogranular platelet syndromes, such as alpha-delta platelet storage pool deficiency. PMID- 17127481 TI - Platelet-mediated erythromelalgic, cerebral, ocular and coronary microvascular ischemic and thrombotic manifestations in patients with essential thrombocythemia and polycythemia vera: a distinct aspirin-responsive and coumadin-resistant arterial thrombophilia. AB - Microvascular circulation disturbances including erythromelalgia, its microvascular ischemic complications, and migraine-like atypical or typical transient ischemic cerebral, ocular, and coronary ischemic attacks are specific clinical manifestations in patients with essential thrombocythemia (ET), and polycythemia vera (PV) associated with thrombocythemia. Thrombocythemia (ET and PV) patients with microvascular disturbances have shortened platelet survival, increased beta-thromboglobulin (beta-tg), platelet factor 4 (PF4), and thrombomoduline (TM) levels, and increased urinary thromboxane B2 (TxB2) excretion indicating platelet-mediated processes in vivo. Inhibition of platelet cyclooxygenase (COX 1) by aspirin is followed by relief of microvascular disturbances, correction of shortened platelet survival, and return of plasma levels of beta-tg, PF4, TM levels and TxB2 excretion to normal. The transient ischemic attacks and thrombotic complications in thrombocythemia are very likely caused by hypersensitive platelets produced by spontaneously proliferating enlarged megakaryocytes in the bone marrow of ET and PV patients. In contrast to normal platelets in healthy individuals the circulating hypersensitive thrombocythemic platelets spontaneously activate and secrete their products, thus forming aggregates that transiently plug the microcirculation, or result in occlusive platelet thrombi in arterioles or small arteries. Clear evidence is presented that the microvascular transient ischemic and occlusive thrombotic complications in thrombocythemia patients are relieved by treatment with aspirin and by reduction of platelet counts to normal (<400 x 109/l), but not by coumadin. In patients with thrombocythemia associated with PV, increased hematocrit and whole blood viscosity aggravate the platelet-mediated microvascular ischemic and thrombotic syndrome of thrombocythemia to major arterial and venous thrombotic complications. Correction of hematocrit and blood viscosity by phlebotomy significantly reduces the major arterial and venous thrombotic complications, but fails to prevent the platelet-mediated microvascular circulation disturbances in PV patients because thrombocythemia persists. Complete relief and prevention of microvascular and major thrombosis in PV patients are obtained by treatment with low-dose aspirin on top of phlebotomy or by treatment with the platelet lowering agents, anagrelide, interferon or hydroxyurea. PMID- 17127482 TI - Effects of different blood components on clot retraction analysed by measuring elasticity with a free oscillating rheometer. AB - Free oscillation rheometry (FOR) using the ReoRox 4 instrument makes it possible, at bedside, to study the coagulation process in blood over time and gives information on clotting time and coagulum elastic properties. In order to find out how various factors influence the FOR analysis we studied the coagulation process and change of elasticity over time in non-anticoagulated and citrated blood samples, plasma samples with various platelet concentrations (0-200 x 109/l) and blood samples with various haematocrit (0-40%). Blood samples supplemented with fibrinogen were analysed to elucidate the importance of fibrinogen on elasticity. The importance of the GPIIb/IIIa receptor on platelets was investigated by comparing the elasticity development in blood samples in presence and absence of a GPIIb/IIIa receptor inhibitor, abciximab. Anticoagulation with citrate did not have major influence on the viscoelastic properties of the coagulum. Increasing number of platelets and increasing fibrinogen concentration resulted in higher elasticity while increasing haematocrit gave lower elasticity. Blood samples with GPIIb/IIIa receptor inhibitor had very low elasticity indicating the importance of functional GPIIb/IIIa receptors. In conclusion we consider FOR to be a useful method to study the elastic properties of the coagulum. Various factors such as the number of red blood cells and platelets as well as the fibrinogen concentration should be taken into consideration when evaluating the results. The ReoRox 4 instrument had excellent measuring range and unusually small artefactual effects on clot elasticity induced by the instrument in comparison with published results on other instruments. PMID- 17127483 TI - Testing various fruits for anti-thrombotic effect: i. Mulberries. AB - Prevention of arterial thrombotic disease has high priority in developed countries. As inappropriate diet predispose to acute thrombotic events, regular intake of an antithrombotic diet may be a convenient and effective way of prevention. The present study was performed to examine antithrombotic effect of mulberry varieties. A shear-induced in vitro platelet reactivity/thrombolysis test (Gorog Thrombosis Test) was used to screen for antiplatelet and thrombolytic activities. In case of effectiveness, it was followed by an in vivo test of laser induced thrombosis in mice. Antioxidant capacity was assessed by superoxide anion and radical scavenging activities. Total polyphenolics, anthocyanin and citrate contents were also measured. The tested varieties showed different effect in vitro on platelet reactivity and endogenous thrombolytic activity. Varieties inhibiting platelet reactivity were antithrombotic in vivo regardless inhibition or enhancement of thrombolysis. Those mulberry varieties, which enhanced platelet reactivity in vitro, were prothrombotic only if inhibitory activity on endogenous thrombolysis coexisted with the platelet effect. Antioxidant activities and polyphenolics content did not affect platelets and the overall thrombotic status. However, antioxidant activities and polyphenolics content significantly correlated with the endogenous thrombolytic activity. These data showed that mulberry varieties can be grouped into subclasses with either anti- or prothrombotic activities. Antioxidant activities and polyphenolic contents do not affect platelets but may enhance endogenous thrombolysis, thus causing an overall antithrombotic effect. PMID- 17127484 TI - Platelet-derived chemokine RANTES may be a sign of restenosis after percutaneous coronary intervention in patients with stable angina pectoris. AB - Inflammation plays a pathogenic role in the development of restenosis after percutaneous coronary intervention (PCI). We measured and compared the ratio of elevated levels of regulated on activation normally T-cell expressed and secreted (RANTES), monocytic chemotactic peptide-1 (MCP-1), soluble (s) P-selectin and sL selectin after PCI. Plasma levels of chemokines and soluble markers were measured before, 1, 3 and 7 days after PCI in 52 patients (43 males and nine females, aged 63 +/- 10 years) who underwent PCI and who had repeated angiograms at a 6-month follow-up. Restenosis occurred in 16 (31%) patients. A significant and time dependent increase in sL-selectin was observed in the restenosis group. However, there were no significant differences in MCP-1 levels with or without restenosis. sP-selectin levels in the restenosis group exhibited a transient elevation at 3 days after PCI. RANTES levels were no different at baseline between patients with or without restenosis. However, a significant and time-dependent decrease in RANTES levels were observed in the non-restenosis group, and patients with restenosis compared with patients without restenosis had a statistically significant ratio of elevated levels of RANTES. These findings suggest that restenosis development after PCI in patients with effort angina pectoris may involve leukocyte activation at an early period after PCI. In addition, platelet derived chemokine RANTES may be a sign of restenosis after PCI in patients with stable angina pectoris. PMID- 17127485 TI - Platelet-derived microvesicles induce differential gene expression in monocytic cells: a DNA microarray study. AB - Platelet-derived microvesicles (PMV) that are shed from the plasma membrane of activated platelets, expose various platelet-type antigens on their surface and are able to adhere to other blood cells and endothelial cells. There are several clinical conditions with markedly increased numbers of PMV, e.g. acute coronary syndrome, thrombotic microangiopathy and sepsis. To prove whether PMV may contribute to an inflammatory response we used DNA microarray technology to study the effect of PMV on gene expression in the prototypic monocytic cell line MonoMac 6 (MM6). PMV were generated by activating human platelets in plasma with collagen and subsequent removal of platelets and plasma by repeated centrifugation. MM6 were incubated for 2 h with PMV in a ratio corresponding to 75 platelets/cell, or saline as control. After RNA isolation, reverse transcription and fluorescence labelling, cDNA was hybridized on a medium density microarray comprising 5308 probes addressing 4868 transcripts of 4730 human genes relevant to inflammation, immune response and related processes. The formation of PMV-MM6 conjugates was associated with significant variations in gene expression, i.e. 93 genes were found to be differentially expressed (P < 0.001; q < 0.087). Among them, 47 genes with annotated transcripts and proteins were identified. Using Ingenuity Pathway Analysis, 37 of the differentially expressed genes were identified as parts of networks associated with functional pathways including cell-to-cell signalling, cellular growth and proliferation, regulation of gene expression and lipid metabolism. For sphingosine kinase-1 the increased expression could be confirmed exemplarily not only by RT-PCR but also on the enzyme activity level. The data indicate that PMV signal differential expression of inflammation-relevant genes in monocytic cells and may represent a novel link between hemostasis and inflammation. PMID- 17127486 TI - Platelet and monocyte activation by hyperglycemia and hyperinsulinemia in healthy subjects. AB - Type 2 diabetes mellitus (T2DM) patients have hyperglycemia and hyperinsulinemia and increased risk of atherosclerosis and acute vascular complications. We have reported elevated circulating tissue factor procoagulant activity (TF-PCA) during hyperglycemia (HG) and hyperinsulinemia (HI) in normal subjects. To evaluate the effect of hyperglycemia and hyperinsulinemia on blood cell activation, we assessed platelet CD40L and P-selectin, monocyte tissue factor (TF), and the formation of monocyte-platelet and neutrophil-platelet aggregates. These were assessed in the resting state and following activation with ADP and thrombin (SFLLRN). Healthy individuals were subjected to 24 h of hyperglycemia and hyperinsulinemia, selective hyperglycemia, selective hyperinsulinemia, or normal glucose and insulin. Platelet CD40L expression increased with high glucose/high insulin, selective hyperglycemia and selective hyperinsulinemia. Monocyte platelet aggregates increased with high glucose/high insulin. Monocyte TF expression increased with high glucose/high insulin and with selective hyperinsulinemia. Upon stimulation with ADP and SFLLRN, monocyte-platelet and neutrophil-platelet aggregates, platelet CD40L and P-selectin, and monocyte TF increased compared to the resting state but was not different between 0 and 24 h, indicating that the responsiveness to those agonists was not altered. CONCLUSIONS: Hyperglycemia-hyperinsulinemia in healthy individuals induced platelet activation and monocyte TF expression promoting a procoagulant and proinflammatory state that may contribute to acute vascular events and atherogenesis. Platelet responsiveness to activation with ADP or SFLLRN appears not to be altered by hyperglycemia-hyperinsulinemia. PMID- 17127487 TI - Lack of association between gene sequence variations of platelet membrane receptors and aspirin responsiveness detected by the PFA-100 system in patients with coronary artery disease. AB - Platelet membrane receptors play a pivotal role in thrombus formation. Expression of platelet membrane receptors are under genetic control and gene sequence variations of receptors pivotal to thrombotic formation have been hypothesized to contribute to different degrees of individual response to aspirin. The aim of the present study was to investigate the impact of functional genetic polymorphisms of platelet membrane receptors on aspirin sensitivity assessed by means of the PFA-100 system in patients with coronary artery disease. Gene sequence variations of three platelet membrane receptors (GPIa/IIa, P2Y12, GPIIb/IIIa) pivotal to thrombus formation were assessed in 76 patients with coronary artery disease on chronic aspirin treatment. Patients with reduced sensitivity to aspirin were defined when closure-times of collagen/epinephrine cartridges < or =193 seconds and coined as PFA-100 non-responders. PFA-100 non-responders were observed in 33% of patients. Patients with diabetes mellitus were more frequently PFA-100 non responders. Closure times of collagen/ADP coated cartridges were reduced in PFA 100 non-responders. The genotype distribution was similar in PFA-100 responder and non-responder patients for all three genotypes and did not vary in contemporaneous carriers of allelic variants. In conclusion, in vitro determined sensitivity to aspirin assessed using PFA-100 is not associated with gene sequence variations of platelet membrane receptors key to thrombus formation. PMID- 17127488 TI - Rosiglitazone-induced immune thrombocytopenia. PMID- 17127490 TI - Insulins and oral hypoglycemic agents in pregnancy. AB - Numerous studies have established a direct relationship between maternal levels of glycemic control and neonatal outcomes for pregnancies complicated by diabetes. The past several years have seen the addition of insulin analogues as well as many new oral agents to the pharmacological armamentarium available to treat diabetes. Insulin analogs (both rapid and long acting) are of potential interest for women with insulin-requiring diabetes because of the improved control reported in non-pregnant individuals. Insulin lispro is the only insulin analog to be systematically studied in pregnancy. At this time, the majority of evidence suggests that insulin lispro does not cross the placenta and does not have adverse maternal or fetal effects during pregnancy in women with diabetes. For women with gestational diabetes mellitus (GDM) and type 2 diabetes, which are characterized by insulin resistance and relatively decreased insulin secretion, treatment with oral hypoglycemic agents is generating much excitement. Most retrospective studies and the published clinical experience have failed to demonstrate an increased risk of neonatal hypoglycemia and other neonatal morbidities with glyburide or metformin. To date there has been only one randomized controlled trial utilizing glyburide, which found it to be safe and effective in the management of GDM. More intensive investigation regarding the safety and feasibility of oral agents in pregnancies complicated by type 2 diabetes is necessary. PMID- 17127491 TI - HCG--A new kid on the block in prematurity prevention. AB - Human chorionic gonadotropin (HCG) is a molecule with multiple endocrine, paracrine, and immunoregulatory actions. Its importance for the enhancement of fertility, successful implantation, and survival of the conceptus in early gestation is recognized. However, studies conducted worldwide in recent years indicate that HCG may also play a significant role in maintaining pregnancy well after the first trimester. Emerging evidence suggests that different biomolecular and physiologic effects of HCG are concordantly directed toward inhibition of myometrial contractility to maintain pregnancy. These studies have prompted preliminary animal and human testing of HCG for the prevention of preterm birth. This article reviews the current knowledge as well as the future perspectives on HCG as a useful new tool in prematurity prevention. PMID- 17127492 TI - Funisitis in term pregnancy is associated with microbial invasion of the amniotic cavity and intra-amniotic inflammation. AB - OBJECTIVE: Funisitis is the histologic counterpart of the fetal inflammatory response syndrome, which is a multisystemic disorder associated with impending preterm delivery and adverse neonatal outcome. The purpose of this study was to examine the relationship between funisitis and the microbiologic status of amniotic fluid (AF) and AF white blood cell (WBC) count in patients at term. METHODS: The relationship between the presence of funisitis, AF culture, and AF WBC count was examined in 832 consecutive patients who delivered a term neonate within 72 hours of amniocentesis. AF was cultured for aerobic and anaerobic bacteria, as well as for mycoplasmas. Funisitis was diagnosed in the presence of neutrophil infiltration into the umbilical vessel walls or Wharton's jelly. AF WBC count was analyzed in a hemocytometer chamber. Nonparametric statistics were used for data analysis. RESULTS: Funisitis was present in 4% (30/832) of cases. A positive AF culture was more common in cases with funisitis than in those without funisitis (17% vs. 5%; p < 0.05). Patients with funisitis had a significantly higher median AF WBC count than those without funisitis (median >1000 cells/mm3 vs. median 2 cells/mm3; p < 0.001). The frequency of funisitis and of a positive AF culture was 1% in women without labor and with intact membranes and the frequencies and the median AF WBC count increased in the presence of labor or rupture of membranes. CONCLUSION: Funisitis is present in 4% of women at term and is associated with microbial invasion of the amniotic cavity (MIAC) and inflammation as reflected by increased AF WBC count. PMID- 17127493 TI - A scoring system for detection of macrosomia and prediction of shoulder dystocia: a disappointment. AB - OBJECTIVE: To develop a scoring system for the detection of a macrosomic fetus (birth weight (BW) >or= 4000 g) and predict shoulder dystocia among large for gestational age fetuses. STUDY DESIGN: We retrospectively identified all singletons with accurate gestational age (GA) that were large for GA (abdominal circumference (AC) or estimated fetal weight (EFW) >or= 90% for GA) at >or=37 weeks with delivery within three weeks. The scoring system was: 2 points for biparietal diameter, head circumference, AC, or femur length >or=90% for GA, or if the amniotic fluid index (AFI) was >or=24 cm; for biometric parameters <90% or with AFI <24 cm, 0 points. The predictive values for detection of shoulder dystocia were calculated. RESULTS: Of the 225 cohorts that met the inclusion criteria the rate of macrosomia was 39% and among vaginal deliveries (n = 120) shoulder dystocia occurred in 12% (15/120; 95% confidence interval (CI) 7-20%). The sensitivity of EFW >or=4500 g to identify a newborn with shoulder dystocia was 0% (95% CI 0-21%), positive predictive values 0% (95% CI 0-46%), and likelihood ratio of 0. For a macrosomia score >6, the corresponding values were 20% (4-48%), 25% (5-57%) and 2.3. CONCLUSION: Though the scoring system can identify macrosomia, it offers no advantage over EFW. The scoring system and EFW are poor predictors of shoulder dystocia. PMID- 17127494 TI - Normal standards of fetal behavior assessed by four-dimensional sonography. AB - OBJECTIVE: In this prospective randomized study, fetal behavior was investigated in order to determine the standard parameters of fetal movements and facial expressions in all three trimesters of normal pregnancy. METHODS: Sixty-three pregnant women with singleton pregnancies in all trimesters were included in the investigation. Four-dimensional (4D) ultrasound was performed for each patient over a 30-minute period. Variables of maternal and fetal characteristics including gestational age, eight fetal movement patterns in the first trimester, and sixteen parameters of fetal movement and fetal facial expression patterns in the second and third trimesters were recorded for the construction of fetal neurological charts. RESULTS: In the first trimester, a tendency towards an increased frequency of fetal movement patterns with increasing gestational age was noticed. Only the startle movement pattern seemed to occur stagnantly during the first trimester (p > 0.05). At the beginning of the second trimester, the frequency of fetal movement patterns tended to increase. During the second and third trimester, multiple regression and polynomial regression revealed statistically significant changes in tongue expulsion (p < 0.05), smiling (p < 0.05), grimacing (p < 0.05), swallowing (p < 0.05), eye blinking (p < 0.01), head movements, and all hand to body contact movements (p < 0.01), except for head anteflexion (p > 0.05). There were no statistically significant changes during the second and third trimesters in mouthing, yawning, and sucking (p > 0.05). At the middle of the third trimester, the fetuses displayed decreasing or stagnant incidence of fetal facial expressions except for eye blinking, which showed increased frequency with increasing gestational age. A statistically significant correlation was found between all head movements and hand to body contact patterns during the second and third trimesters except for head anteflexion (r = 0.231; p > 0.05). CONCLUSIONS: The full range of quantitative fetal facial expressions and fetal movement patterns can be assessed successfully by 4D sonography. It is important to be able to assess normal fetal behavior throughout gestation to identify abnormal behavior before birth. PMID- 17127495 TI - Is severe perineal damage increased in women with prior anal sphincter injury? AB - OBJECTIVE: There is conflicting data in the literature regarding the risk of obstetric anal sphincter laceration in patients with a prior laceration. This retrospective chart review seeks to examine the risk of recurrence of obstetric anal sphincter lacerations. METHODS: Patients who sustained anal sphincter laceration at delivery during a 13-year time period from January 1991 to December 2003 were identified from the medical records database at Temple University Hospital. All subsequent deliveries in this group of patients were extracted from the database. Chart review was performed on all subsequent deliveries with specific attention to demographic factors such as age, race, parity, etc., maternal weight, fetal weight, presence of maternal diabetes, and labor characteristics such as induction or augmentation of labor, instrumentation at delivery (vacuum or forceps), use of episiotomy, and degree of perineal laceration. RESULTS: There were 23 451 vaginal deliveries at Temple University Hospital between January 1, 1991 and December 31, 2003. Anal sphincter laceration was noted in 778 subjects. Subsequent deliveries among the group of patients with prior sphincter tears numbered 271. Six (2.4%) patients had recurrence of anal sphincter lacerations, and five of them were third degree lacerations. The rate of recurrent lacerations was not significantly different from the rate of initial lacerations (2.4% vs. 3.3%; odds ratio 0.72, 95% confidence interval 0.33-1.59; p = 0.4). Women who sustained recurrent lacerations were older, more obese (mean weight 92 kg vs. 82 kg), had larger babies (3506 g vs. 3227 g), and were more likely to have episiotomies (66.7% vs. 7%) or instrumental deliveries (33.3 vs. 6.5%). CONCLUSION: Prior anal sphincter laceration does not result in an increased rate of recurrence. Operative vaginal delivery particularly with episiotomy is a risk factor for both initial and recurrent laceration. PMID- 17127496 TI - Screening or diagnostic: markedly elevated glucose loading test and perinatal outcomes. AB - OBJECTIVE: To determine the diagnostic value of markedly elevated 50-g glucose loading test results (>or=200 mg/dL) and associated perinatal outcomes. METHOD: This was a retrospective cohort study of 14 771 pregnancies screened for gestational diabetes mellitus (GDM) between 1988 and 2001. The positive predictive value of the 50-g oral glucose loading test (GLT) results as measured by plasma glucose value was examined. Perinatal outcomes were assessed for women with GLT results >or=200 mg/dL compared to GLT <200 mg/dL, stratified by the diagnosis of GDM. Statistical comparisons were made using the Chi-square test and Student's t-test and potential confounding factors were controlled for using multivariable logistic regression analyses. A p value <0.05 and 95% confidence intervals were used to indicate statistical significance. RESULTS: The positive predictive values for a GDM diagnosis were 62% for GLT results between 180 and 189 mg/dL, 79% for those between 200 and 209 mg/dL, and 100% for GLT results >or=230 mg/dL. Compared to women with a GLT result <200 mg/dL, among women not diagnosed with GDM but with a GLT >or=200 mg/dL the adjusted odds ratio (aOR) for cesarean delivery was 4.18 (95% confidence intervals, 1.15-15.2). These women also had higher aORs for preterm delivery <32 weeks (aOR = 8.05 (1.02-63.6)), shoulder dystocia (aOR = 15.14 (1.64-140)), and their neonates were more likely to have a 5-minute Apgar score <7 (aOR = 6.41 (1.23-33.3)). For women diagnosed with GDM and with a GLT >or=200 mg/dL, the aOR for cesarean delivery was also elevated compared to those with a GLT <200 mg/dL (aOR = 2.24 (1.19-4.21)). CONCLUSION: A GLT value of >or=200 mg/dL is not absolutely diagnostic for gestational diabetes but is associated with unfavorable perinatal outcomes. PMID- 17127497 TI - Terbutaline inhibits corticotropin-releasing hormone (CRH) expression in human trophoblast cells. AB - OBJECTIVE: To evaluate the effect of beta-adrenergic agonists on the regulation of the expression of the placental corticotropin-releasing hormone (CRH) gene. STUDY DESIGN: Term placentae were collected at the time of elective cesarean section, and trophoblast cells were harvested, isolated, and cultured. The isolated trophoblasts were plated, cultured and subsequently treated with cortisol, terbutaline, RU486, or vehicle control. CRH expression, mRNA abundance of CRH, and the housekeeping gene beta-actin were evaluated by Northern blot analysis. RESULTS: Exposure of the trophoblasts to terbutaline (10(-8) M) inhibited the expression of the CRH gene as depicted by Northern blot analysis. Co-addition of terbutaline (10(-8) M) and RU486 (10(-6) M) did not block the stimulatory effects of RU486 on placental trophoblast cells. CONCLUSION: The beta adrenergic agonist terbutaline inhibits the expression of CRH in human trophoblasts. This finding may provide insight into the mechanism of action of terbutaline as a tocolytic agent. PMID- 17127498 TI - Use of inferior vena cava filters in thromboembolic disease during labor: case report with a literature review. AB - Thromboembolic diseases during pregnancy are usually managed by conventional anticoagulation and patients are at high risk of pulmonary embolism. Inferior Vena Cava (IVC) filters can be used in cases of documented pulmonary embolism (PE) where anticoagulation is contraindicated or has failed. In our case the patient was diagnosed as having a deep vein thrombosis (DVT) and was started on anticoagulant therapy. Twenty four hours afterwards she went into labor and an IVC filter was inserted due to the risk of pulmonary embolism. She was managed successfully during labor and postpartum period. This shows that IVC filters can be used during labor to try and prevent pulmonary emboli. PMID- 17127499 TI - Diffuse congenital cystic lung disease with spontaneous regression. PMID- 17127500 TI - Heterotopic pregnancy: incidental finding at a term cesarean delivery. PMID- 17127501 TI - History of intrauterine curettage and placental complications. PMID- 17127502 TI - Cytotoxic compounds of Physena madagascariensis from the Madagascar rain forest paragraph sign. AB - Two new flavanones, remangiflavanones D and E (1 and 2), were isolated from an extract of the twigs, leaves, and flowers of Physena madagascariensis together with three known flavanones, remangiflavanones A-C (3-5), and (E)-N feruloyltyramine (6). The structures of the new compounds 1 and 2 were established on the basis of one-dimensional and two-dimensional NMR spectroscopic data interpretation. All compounds were evaluated for their cytotoxicity in the A2780 human ovarian cancer cell line. Compound 5 was the most active with an IC50 value of 2.5 microg mL-1. PMID- 17127503 TI - Cilistepoxide and cilistadiol, two new withanolides from Solanum sisymbiifolium. AB - In addition to the known cilistol A, two new withanolides have been isolated from leaves and stem of Solanum sisymbiifolium, and assigned the structures 1-oxo-5,6; 22,26; 24,25-triepoxy-17,26-dihydroxyergost-2-ene and 1-oxo-22,26; 24,25-diepoxy 5,6,17,26-tetrahydroxy ergost-2-ene, namely cilistepoxide and cilistadiol. PMID- 17127504 TI - Constituents of Psiadia terebinthina A.J. Scott, an endemic Asteraceae from Mauritius. AB - Kaemferol-3-methyl ether (1), quercetin-3-methyl ether (2), kaemferol-3,7 dimethyl ether (3), 3-caffeoyl quinic acid (4) and 3,4-O-dicaffeoyl quinic acid (5) have been isolated for the first time from the leaves of Psiadia terebinthina A.J. Scott (Asteraceae). The identity of the compounds 1-5 were confirmed by various spectroscopic methods. PMID- 17127505 TI - Viscum album aqueous extract induces NOS-2 and NOS-3 overexpression in Guinea pig hearts. AB - Viscum album L. aqueous extract, on the Langendorff isolated and perfused heart model, decreases coronary vascular resistance, when compared to control group (36.00 +/- 2.00 vs. 15.80 +/- 1.96 dyn s cm-5). Our data support the fact that this mechanism involves NOS-2 and NOS-3 overexpression (4.65 and 7.89 times over control, respectively), which is correlated with increases in NO (6.24 +/- 2.49 vs. 147.95 +/- 2.79 pmol) and cGMP production (43.94 +/- 2.00 vs. 74.81 +/- 1.96 pmol mg-1 of tissue), compared to control values. Such an effect is antagonized by gadolinium(III) chloride, L-NAME and ODQ. Therefore, coronary vasodilator effect elicited by V. album L. aqueous extract is mediated by the NO/sGC pathway. PMID- 17127506 TI - 5alpha-Ergost-24(28)-ene-3,6-dione - new steroid from the Pacific sponge Geodinella robusta. AB - A new diketosteroid, 5alpha-ergost-24(28)-ene-3,6-dione (1), has been isolated from the sponge Geodinella robusta. Its structure was determined by combined spectroscopic methods and chemical evidence. PMID- 17127507 TI - A new cerebroside from fruits of Ailanthus altissima Swingle. AB - A new cerebroside and three known cycloartan triterpenes were isolated from fruits of Ailanthus altissima Swingle. Their structures were identified as 1-O beta-D-glucopyranosyl-(2S, 3R, 4E, 9E)-2-(2'R-hydroxyhexadecenoy)-4, 9 octadecadiene-1, 3-diol (1), 9, 19-cyclolanost-23 (Z)-ene-3beta, 25-diol (2), cycloart-25-ene-3beta, 24R-diol (3), and cycloart-25-ene-3beta, 24S-diol (4) by means of chemical and spectroscopic analysis. Compounds 2, 3, and 4 were isolated from genus Ailanthus for the first time. The analgesic activity of 1 was also evaluated. PMID- 17127508 TI - A new flavonoid from Chromolaena odorata. AB - Fractionation of the aerial parts of Chromolaena odorata afforded a new flavonoid, 5,7-dihydroxy-6,4'-dimethoxyflavanone, in addition to 14 known flavonoid compounds, six of which had not been isolated previously from this plant. The structure of the new compound was determined by spectroscopic methods particularly 2D-NMR analysis. PMID- 17127509 TI - Chemical constituents from Phoebe minutiflora II. AB - Two new lignans were isolated from Phoebe minutiflora, namely 8' epiaristotetralone (1) and 8'-epiaristoligone (2), together with seven known compounds. Their structures and absolute stereochemistry were determined by spectral analysis (NMR and CD) and chemical correlation. PMID- 17127510 TI - One new anthraquinone from marine Streptomyces sp. FX-58. AB - One new anthraquinone, 1,8-dihydroxy-2-ethyl-3-methylanthraquinone (1), together with two known compounds octadecanoic acid (2) and cholest-4-en-3-one (3) was isolated from marine actinomycete Streptomyces sp. FX-58. The structure of 1 was elucidated on the basis of spectroscopic methods, especially, the 2D-NMR spectral analysis. The cytotoxic activities of 1 were evaluated in vitro. PMID- 17127511 TI - A new quassinoid, ailantinol H, from Ailanthus altissima. AB - A new quassinoid, ailantinol H, was isolated from the aerial parts of Ailanthus altissima. The structure was elucidated based on spectral evidence. PMID- 17127512 TI - Antioxidant oligomeric proanthocyanidins from Cistus salvifolius. AB - The purified proanthocyanidin oligomers of Cistus salvifolius herb extract accounted for 78% of the total proanthocyanidins and 73% of the total antioxidant activity of this extract. To elucidate the structure of the oligomer, it was depolymerized by acid catalysis in the presence of phloroglucinol. The structures of the resulting flavan-3-ols and phloroglucinol adducts were determined on the basis of 1D- and reverse 2D-NMR (HSQC, HMBC) experiments of their peracetylated derivatives, MALDI-TOF-MS and CD spectroscopy. These observations resulting from the degradation with phloroglucinol were confirmed by 13C NMR spectroscopy of the oligomer. The mean molecular weight of the higher oligomeric fraction was estimated to be 5-6 flavan-3-ol-units. PMID- 17127513 TI - Isolation and structural determination of xerophytolic acid A, a 3-geranyl-4 hydroxybenzoate derivative from Xerophyta plicata. AB - A novel compound of a rare class of secondary metabolites, 3-geranyl-4 hydroxybenzoic acid derivative, methyl xerophytolate A (2), was isolated from the crude extract of Xerophyta plicata (Velloziaceae) collected in Rio de Janeiro, Brazil. The structure of 2 was elucidated using spectroscopic methods. PMID- 17127514 TI - A new kaempferol diglycoside from Datura suaveolens Humb. & Bonpl. ex. Willd. AB - A new flavonol glycoside, kaempferol 3-O-alpha-L-arabinopyranosyl-7-O-beta-D glucopyranoside (1), has been isolated from methanol extract of leaves of Datura suaveolens (Solanaceae), along with six other known compounds, which include kaempferol 3-O-alpha-L-arabinopyranoside (2), 3-phenyl lactic acid, 3-(3-indolyl) lactic acid, and its methyl ester, physalindicanol A and physalindicanol B. The structural elucidation of 1 and characterization of the known compounds are based on detailed spectral analysis (ESI-FTICR-MS and 2D-NMR). This is the first report of isolation of these compounds from this plant. PMID- 17127515 TI - Two new xanthones from Hypericum japonicum. AB - Two new xanthones, 1,6-dihydroxyisojacereubin-5-O-beta-D-glucoside (1) and 3,6,7 tri-hydroxy-1-methoxy-xanthone (2), were isolated from Hypericum japonicum. The structural elucidation of the isolated compounds were primarily based on HREIMS, EIMS, UV, IR, 1D-, and 2D-NMR analyses, including COSY, HMQC, HMBC, and NOESY correlations. PMID- 17127516 TI - A novel bieremophilanolide from Ligularia lapathifolia. AB - A new bieremophilanolide was isolated from the roots and rhizomes of Ligularia lapathifolia. Its structure was established as 8,8'-bi-3beta-angeloyloxy eremophil-7(11)-en-12,8alpha(14beta,6alpha)-diolide (1) by IR, MS, 1D, and 2D NMR experiments. PMID- 17127517 TI - A new gamma-lactone from the leaves of Cinnamomum kotoense. AB - A new C19 gamma-lactone, cinnakotolactone (1), along with a known analogue, isolinderanolide B (2), were isolated from the n-hexane layer of the leaf extracts of Cinnamomum kotoense. Their structures were elucidated on the basis of spectroscopic analysis. The anti-proliferation activities of 1 and 2 were evaluated against human HT29 and MCF-7 cancer cell lines, and their IC50 values ranged from 3.3 +/- 0.3 to 25.8 +/- 5.3 microM. PMID- 17127518 TI - Two new xanthone diglycosides from Swertia longifolia Boiss. AB - From aerial parts of Swertia longifolia Boiss., which grows in the north of Iran, five xanthones, two of which in diglycosidic form, were isolated. The structures were confirmed by means of their spectral data as isobellidifolin, bellidin, gentisein, 1,5-dihydroxy-3-methoxy-6-O-primeverosyl xanthone, and 8-hydroxy-3,5 dimethoxy-1-O-primeverosyl xanthone, the latter two of which were new derivatives in the plant kingdom. PMID- 17127519 TI - Influence of various diet regimens on deterioration of hepatic function and hematological parameters following carbon tetrachloride: a potential protective role of natural honey. AB - The objective was to assess the effects of commercial regular diet as control, total food restriction with honey, commercial regular diet with dextrose, or total food restriction with dextrose, on blood variables after carbon tetrachloride (CCl4) administration. Sprague Dawley albino rats were divided into four groups, 10 rats each; Group 1 rats were on commercial regular diet, Group 2 rats were on commercial regular diet with 50% dextrose, Group 3 rats were on total food restriction with 50% dextrose, and Group 4 rats were on total food restriction with 50% honey. Rats in all the groups were i.m. administered CCL4 (2.4 mL kg b. wt.-1). Blood tests including ALT, AST, serum albumin, serum protein, BUN, blood glucose (BG), hemoglobin (Hb), and white blood cell (WBC) were performed before CCl4 administration and repeated after 48 and 96 h of post injection. In Group 1, CCl4 caused significant elevation in AST and ALT, and decrease in BS, WBC, and BUN; lower elevation in AST and ALT at 48 h and decreased AST and ALT at 96 h were obtained when dextrose was added to commercial regular diet (Group 2). Using dextrose alone (Group 3), though there was significant elevation of AST and ALT and decrease in BUN and WBC as compared to baseline values, significant decrease in ALT, AST, and BUN as compared to control was obtained. During absolute honey feeding (Group 4), elevation in AST and ALT obtained, following CCl4 administration was significantly less than the values obtained in all other groups; with lower elevation in AST and ALT as compared to baseline values. Honey increased serum albumin, serum protein, BG, and caused lower reduction in Hb. Conclusively, exclusive honey feeding (50% concentration) significantly modifies and ameliorates biochemical and hematological changes obtained after CCl4 injection. PMID- 17127520 TI - Two new eremophilanolides from Xanthium sibiricum. AB - Two new eremophilanolides, sibiriolides A (1) and B (2), were isolated from the aerial parts of Xanthium sibiricum. The structures of the new compounds were identified as 4S,5R,7R,8R, 11R-2-oxo-1(10)-eremophilen-12,8-olide (1) and 4S,5R,7R,8R,11S-2-oxo-1(10)-eremophilen-12,8-olide (2) by HREIMS and NMR spectroscopic techniques in combination with X-ray crystallographic analysis and CD measurements. PMID- 17127521 TI - Sterols isolated from Tubifex tubifex. AB - Four 3beta-hydroxysterols isolated from chloroform extract from Tubifex tubifex were shown to have the structures stigmast-23-en-3-ol, stigmast-7,14-dien-3-ol, 22-dehydrocholesterol, and 24 methylenecholesterol based on spectroscopic methods. Stigmasterol and beta-sitosterol were also investigated on the basis of mass spectral analyses and compared with known compounds. PMID- 17127523 TI - Genetics of antimicrobial resistance. AB - Antimicrobial resistant strains of bacteria are an increasing threat to animal and human health. Resistance mechanisms to circumvent the toxic action of antimicrobials have been identified and described for all known antimicrobials currently available for clinical use in human and veterinary medicine. Acquired bacterial antibiotic resistance can result from the mutation of normal cellular genes, the acquisition of foreign resistance genes, or a combination of these two mechanisms. The most common resistance mechanisms employed by bacteria include enzymatic degradation or alteration of the antimicrobial, mutation in the antimicrobial target site, decreased cell wall permeability to antimicrobials, and active efflux of the antimicrobial across the cell membrane. The spread of mobile genetic elements such as plasmids, transposons, and integrons has greatly contributed to the rapid dissemination of antimicrobial resistance among several bacterial genera of human and veterinary importance. Antimicrobial resistance genes have been shown to accumulate on mobile elements, leading to a situation where multidrug resistance phenotypes can be transferred to a susceptible recipient via a single genetic event. The increasing prevalence of antimicrobial resistant bacterial pathogens has severe implications for the future treatment and prevention of infectious diseases in both animals and humans. The versatility with which bacteria adapt to their environment and exchange DNA between different genera highlights the need to implement effective antimicrobial stewardship and infection control programs in both human and veterinary medicine. PMID- 17127524 TI - Genetic linkage and horizontal gene transfer, the roots of the antibiotic multi resistance problem. AB - Bacteria carrying resistance genes for many antibiotics are moving beyond the clinic into the community, infecting otherwise healthy people with untreatable and frequently fatal infections. This state of affairs makes it increasingly important that we understand the sources of this problem in terms of bacterial biology and ecology and also that we find some new targets for drugs that will help control this growing epidemic. This brief and eclectic review takes the perspective that we have too long thought about the problem in terms of treatment with or resistance to a single antibiotic at a time, assuming that dissemination of the resistance gene was affected by simple vertical inheritance. In reality antibiotic resistance genes are readily transferred horizontally, even to and from distantly related bacteria. The common agents of bacterial gene transfer are described and also one of the processes whereby nonantibiotic chemicals, specifically toxic metals, in the environment can select for and enrich bacteria with antibiotic multiresistance. Lastly, some speculation is offered on broadening our perspective on this problem to include drugs directed at compromising the ability of the mobile elements themselves to replicate, transfer, and recombine, that is, the three "infrastructure" processes central to the movement of genes among bacteria. PMID- 17127525 TI - Reservoirs of antibiotic resistance genes. AB - A potential concern about the use of antibiotics in animal husbundary is that, as antibiotic resistant bacteria move from the farm into the human diet, they may pass antibiotic resistance genes to bacteria that normally reside in a the human intestinal tract and from there to bacteria that cause human disease (reservoir hypothesis). In this article various approaches to evaluating the risk of agricultural use of antibiotics are assessed critically. In addition, the potential benefits of applying new technology and using new insights from the field of microbial ecology are explained. PMID- 17127526 TI - Antimicrobial resistance in Scandinavia after ban of antimicrobial growth promoters. AB - The banned use of antimicrobial growth promoters resulted in a considerably decreased use of antimicrobials in food animal production in Sweden (65%), Denmark (47%), Norway (40%) and Finland (27%). The current prevalence of antimicrobial resistance in animal bacterial populations is also considerably lower than in some other countries in the EU. In the swine production, no or limited effect was found in the finisher production (>25 to 30 kg). Temporary negative effects occurred during the post weaning period (7-30 kg). In Denmark, the cost of production from birth to slaughter per pig produced increased by approximately 1.0 euro with a high variability between pig producers. In the broiler production the termination had no significant negative effect on animal health and welfare or on production economy. PMID- 17127527 TI - Tetracycline residues and tetracycline resistance genes in groundwater impacted by swine production facilities. AB - Antibiotics are used at therapeutic levels to treat disease; at slightly lower levels as prophylactics; and at low, subtherapeutic levels for growth promotion and improvement of feed efficiency. Over 88% of swine producers in the United States gave antimicrobials to grower/finisher pigs in feed as a growth promoter in 2000. It is estimated that ca. 75% of antibiotics are not absorbed by animals and are excreted in urine and feces. The extensive use of antibiotics in swine production has resulted in antibiotic resistance in many intestinal bacteria, which are also excreted in swine feces, resulting in dissemination of resistance genes into the environment. To assess the impact of manure management on groundwater quality, groundwater samples have been collected near two swine confinement facilities that use lagoons for manure storage and treatment. Several key contaminant indicators - including inorganic ions, antibiotics, and antibiotic resistance genes - were analyzed in groundwater collected from the monitoring wells. Chloride, ammonium, potassium, and sodium were predominant inorganic constituents in the manure samples and served as indicators of groundwater contamination. Based on these analyses, shallow groundwater has been impacted by lagoon seepage at both sites. Liquid chromatography-mass spectroscopy (LC-MS) was used to measure the dissolved concentrations of tetracycline, chlortetracycline, and oxytetracycline in groundwater and manure. Although tetracyclines were regularly used at both facilities, they were infrequently detected in manure samples and then at relatively trace concentrations. Concentrations of all tetracyclines and their breakdown products in the groundwater sampled were generally less than 0.5 microg/L. Bacterial tetracycline resistance genes served as distinct genotypic markers to indicate the dissemination and mobility of antibiotic resistance genes that originated from the lagoons. Applying PCR to genomic DNA extracted from the lagoon and groundwater samples, four commonly occurring tetracycline (tet) resistance genes tet(M), tet(O), tet(Q), and tet(W) - were detected. The detection frequency of tet genes was much higher in wells located closer to and down-gradient from the lagoons than in wells more distant from the lagoons. These results suggested that in the groundwater underlying both facilities tetracycline resistance genes exist and are somewhat persistent, but that the distribution and potentially the flux for each tet gene varied throughout the study period. PMID- 17127528 TI - The application of biotechnical and epidemiologic tools for pig health. AB - Health maximization is a farm essential because of its implications for animal well-being and for production. Health care in all livestock species is a dynamic, technologically-limited, and resource-constrained enterprise. As in any health endeavor, challenges abound. Fundamentally, the cornerstones of animal health are good husbandry and good housing. Effective health systems, however, incorporate other management aspects, including biotechnology and epidemiology. The degree to which each affects the incidence and severity of pig disease is not fully appreciated. Shortfalls in management, resource allocation, available tools, and/or understanding are not uncommon. This article reviews the dynamics of pig health emphasizing how technology and management marginally affect the course of disease. Further, management techniques that incorporate technology in disease control, elimination, and eradication are reviewed. Antimicrobials are a key biotechnical tool for maximizing pig health. PMID- 17127529 TI - Health management with reduced antibiotic use - experiences of a Danish pig vet. AB - During the late 1990s, all use of antibiotic growth promotors for production animals was gradually banned in Denmark. At the same time, the legislation on prescription of medicine was tightened and registration of usage became mandatory. Following the ban, the prevalence of diarrhoea - often caused by Lawsonia intercellularis - increased, which led to an expected increased use of therapeutic antibiotics. However, the consumption never reached the same amount of antibiotics as distributed before the ban. As a consequence, production performance dropped with a reduction of 18 g to 50 g per day in daily gain and mortality increased by an extra 0.6% to 1.5%, especially in the weaner period. The cost of this is at least from 1 to 3 US dollars per animal. No reliably substitute to antibiotic has been found yet, but improvement of the diet and management on all levels are important factors for compensation. Further investigations in this area are called for in the future. PMID- 17127530 TI - Health management with reduced antibiotic use - the U.S. experience. AB - Since World War II the use of antimicrobial products associated with food animal production has increased. Antimicrobials along with evolving production practices have significantly increased throughput, animal welfare, and improved health. Concerns surrounding the growing significance of emerging and in some cases rapidly disseminating antibiotic (antimicrobial) resistant bacterial pathogens among human and livestock populations has stimulated a reassessment of this application. The negative publicity has led many consumers and activist groups to believe that protein derived from food animals grown in the absence of those drugs is safer than products derived from the conventionally reared. There is a general fear that antimicrobial usage in agriculture threatens the sustainability of human therapeutic agents and the public wellbeing. The issue has gradually emerged from "fringe group paranoia" to mainstream - finally impacting consumer choices. Antimicrobial resistance concerns have stimulated a significant reaction by the US animal agriculture industry. Numerous pig production entities, large and small, have attempted to create additional pork product value by developing niche marketing opportunities. Thus far most of the subtherapeutic in-feed antimicrobial reduction has been voluntary in the US. Two production areas have developed where reduced usage occurs. First is the growth of antibiotic free production (ABF) and second is an increased use of treatment levels which avoids subtherapeutic criticism. The bulk of this article is directed at new production practices, pig health management, disease elimination, and biosecurity efforts that result from early industry attempts at reduced or excluded antimicrobial pig production. Raising antimicrobial (antibiotic) free (ABF) pork from birth is challenging for a variety of reasons. Some of these challenges can be cost effectively dealt with while others are difficult if not impossible to control in modern production environments. Healthy pig production is essential for sustainable ABF operations. PMID- 17127531 TI - Reduced use of antibiotic growth promoters in diets fed to weanling pigs: dietary tools, part 1. AB - Antibiotics improve health and productive performance of pigs. There is increasing evidence that other dietary ingredients may provide similar, but probably not identical, benefits. A properly prepared immune egg product, spray dried plasma, or a high level of dietary zinc can provide protection against enteric infection in young pigs. Spray-dried plasma also increases growth rate dramatically, and zinc increases it substantially. The evidence in refereed publications also indicates that organic acids increase growth rate substantially, but does not clearly identify the most effective acid or combination of acids. Whey proteins, lactose, mannan oligosaccharide, or a high level of dietary copper provides a modest but useful increase in growth rate. It is not clear whether conventional egg products or other milk proteins may be beneficial. There are many other feed ingredients commercially available that may improve health and productive performance of pigs. PMID- 17127532 TI - Reduced use of antibiotic growth promoters in diets fed to weanling pigs: dietary tools, part 2. AB - Diets formulated to maximize performance of weanling pigs need to support the development of intestinal tissue, support intestinal colonization with beneficial, mainly lactic acid-producing bacteria, and support development of the intestinal and overall immune system. This objective is not likely to be achieved using one single strategy, but there is strong evidence that diets formulated with cereal grains other than corn, with a low concentration of crude protein and with the use of direct-fed microbials, will improve intestinal health and performance of weanling pigs. Further improvements may be observed if the grain part of the diet is fermented prior to feeding or if the diet is fed in a liquid form, but the need for specialized equipment limit the implementation of this strategy. Dietary supplements such as essential oils and nucleosides or nucleotides may also be useful, but more research is needed to verify the effects of these substances. PMID- 17127533 TI - The Take Care program and responsible use of antibiotics. AB - While the contribution of antimicrobial use in pork production to resistant bacteria in human health is uncertain, pork producers have the responsibility to deliver safe and wholesome pork to consumers. The development of a comprehensive producer education and awareness program focused on the responsible use of antimicrobials in pork production is described. The Take Care - Use Antibiotics Responsibly program is based on principles and guidelines that provide the producer and veterinarian the basis for antibiotic use decision-making. The goal of the program is to protect public health while preserving animal health and welfare. PMID- 17127534 TI - Antibiotic use in animal agriculture: what have we learned and where are we going? AB - Antibiotics are enormously important for the humane and efficient production of food animals. These benefits are somewhat offset by the human and animal health antibiotic resistance risks posed by their use in animals. This article provides an overview of what we have learned about antibiotic resistance as an issue in animal agriculture and where that knowledge could lead us in the future. To preserve the effectiveness of antibiotics, more action is needed to ensure their prudent use, particularly in the case of antibiotic growth promoters and antibiotics deemed critically important for human and animal health. PMID- 17127537 TI - Is caffeine addictive?--a review of the literature. AB - The common-sense use of the term addiction is that regular consumption is irresistible and that it creates problems. Caffeine use does not fit this profile. Its intake does no harm to the individual or to society and its users are not compelled to consume it. Though cessation of regular use may result in symptoms such as headache and lethargy, these are easily and reliably reversed by ingestion of caffeine. Some have argued that continued caffeine use is an attempt to suppress low grade withdrawal symptoms such as sleepiness and lethargy. In some moderate users, this is possible; however, in experimental contexts, the phenomenon is too inconsistent to constitute a reliably valid syndrome. PMID- 17127538 TI - Behavioral therapy to augment oral naltrexone for opioid dependence: a ceiling on effectiveness? AB - The effectiveness of antagonist maintenance with oral naltrexone for opioid dependence has been limited by high dropout rates. Behavioral Naltrexone Therapy (BNT) was developed to improve retention on oral naltrexone by integrating voucher incentives, Motivational and Cognitive Behavioral therapies, and a significant other for monitoring medication adherence. In a 6-month, randomized, controlled trial in heroin dependent patients, BNT (N = 36) improved retention in treatment compared to a standard treatment control (Compliance Enhancement (CE); N = 33) (log rank = 4.28; p = .04). Most patients retained beyond 3 months achieved abstinence from opioids, but retention at 6 months was only 22% on BNT and 9% on CE. A systematic review of related controlled trials revealed similar effect sizes in the small to medium range, and substantial dropout. There may be a limit on the extent to which behavioral therapy can overcome poor adherence to oral naltrexone. Future research should consider combinations of behavioral methods with new long-acting injectable or implantable naltrexone formulations. PMID- 17127539 TI - College binge drinking: deviant versus mainstream behavior. AB - College binge drinking is examined from the perspectives of two cultures. The traditional culture views binging as deviant; the second culture promotes it. In this context, logit regression is used to explore the effects of various factors, including student employment and parental education. Employed students are less likely to binge than are students who are not employed. Also, students whose mother is a college graduate, but whose father is not, are more likely to binge than other students. The prescriptions for reducing binge drinking are different when the behavior is perceived as mainstream rather than deviant. The research calls for the development of a process for promoting cultural change in an environment of continually changing student leadership. PMID- 17127540 TI - Spending of remuneration by subjects in non-treatment drug abuse research studies. AB - BACKGROUND: This study examines remuneration spending by drug-using participants in residential drug abuse research. METHODS: Ninety-four adult males who participated in residential, non-treatment drug abuse research studies earned remuneration based on length of stay and specific research procedures. Remuneration could be in cash after discharge or for in-kind purchases and bill payments. Spending of remuneration was extracted from charts and evaluated with multivariate analyses. RESULTS: Participants received average remuneration of 1,454 dollars, taking 59% in cash. Other categories included cigarettes (60.6% of subjects), toiletries (60.6%), clothing (54.3%), and housing (52.1%). Primary drug of abuse, total remuneration, monthly income, length of stay on the residential research unit, age, and education were significantly associated with in-kind remuneration choices. Less total remuneration, intoxication in the month prior to study, higher IQ, and non-white race were associated with taking more in cash. CONCLUSION: Residential drug abuse research participants prefer cash to in kind research remuneration, and their choices reflected drug use and economic status. PMID- 17127541 TI - Behavior therapy for depression in drug dependence (BTDD): results of a stage Ia therapy development pilot. AB - Limited access to positive reinforcers is a central feature in behavioral formulations of substance use and depression, and evidence suggests both disorders share similar environmental contexts. The Behavioral Therapy for Depression in Drug Dependence (BTDD) was developed to target the density of potential reinforcers in a patient's environment to reduce both depression and illicit substance use using therapeutic techniques from three operant based treatment programs, Community Reinforcement Approach, Changing Reinforcement Events, and Treatment-plan Contingency Management. Results of an uncontrolled Stage Ia trial (n = 29), indicated 48% of the participants demonstrated at least a 50% reduction in baseline depression scores during the 16-session treatment program. Those designated as treatment responders completed more out-of-session behavioral activities, attended more treatment sessions, and demonstrated less benzodiazepine use during the program than non-responders. There were no changes in opiate and cocaine use. BTDD may be a useful adjunct to methadone maintenance for treating comorbid depressive disorders. PMID- 17127542 TI - Ethnic origin of alcoholics admitted to an Israeli treatment center. AB - Jews have a low rate of alcohol use and abuse as reported in several different countries. In Israel over the last 10 years there has been a rising rate of alcoholism. We studied consecutive new admissions to our inpatient alcohol center, and an age and sex matched comparison group of patients consecutively admitted with schizophrenia for whether each of their four grandparents was Jewish, or non-Jewish. A significantly higher percentage of alcohol-related admissions were immigrants from the Former Soviet Union (FSU) than among schizophrenics. Among the alcohol-related admissions from the FSU, there were significantly fewer Jewish grandparents than among schizophrenia patients from the FSU. These data could support the concept that biological Jewish ethnicity has a protective effect against alcohol abuse, but are also consistent with cultural transmission of Jewish attitudes toward alcohol use. PMID- 17127543 TI - Introducing evidence-based practices into substance abuse treatment using organization development methods. AB - BACKGROUND: Dissemination of evidence-based practices (EBPs) in addiction settings is a national priority. We tested Organization Development (OD) methods for dissemination. METHODS: Using OD in two addiction treatment programs we developed an organization-specific treatment plan using employee work teams with the goals of changes in organizational policies and procedures and improvement in practitioner skills. RESULTS: OD was effectively applied, but EBPs were premature for these addiction programs because they first needed to address more fundamental aspects of client-clinician interaction and agency treatment philosophy. CONCLUSION: The OD approach in addiction treatment is complementary to other technology transfer efforts by being: (a) "organization-centered," engaging practitioners at all levels; (b) "needs-focused," addressing concerns of the particular organization; (c) flexible in its responsiveness to readiness for change; and (d) relatively affordable. However, before absorbing EBPs, substance abuse treatment organizations must develop strengths in delivering fundamental aspects of care. PMID- 17127544 TI - Variations in drinking patterns in the rural South: joint effects of race, gender, and rural residence. AB - OBJECTIVES: To understand the relative contribution of gender, race (African American vs. Caucasian), and rural residence on variations in drinking patterns, including past year abstinence, at-risk drinking, and recent drinking quantity and frequency for drinkers only. METHODS: A brief health survey was administered by telephone to a probability sample of 11,529 residents of six southern states, over-sampling rural inhabitants. RESULTS: Drinking patterns varied by gender, race, and rural residence in bivariate analysis. Gender effects were independent of rural residence, but race effects on abstinence and at-risk drinking were found only in urban residents and race differences in drinking quantity only in rural residents. Multivariate analysis, controlling for age and education, found gender and rural residence to be the strongest predictors, as well as being an African-American female. CONCLUSIONS: Female gender, African-American race, and rural residence appear protective for at-risk drinking but rural residence dominates racial differences. PMID- 17127545 TI - Evidence-based treatment for opiate-dependent clients: availability, variation, and organizational correlates. AB - The majority of opiate-dependent clients entering substance abuse treatment are referred to "drug-free" (non-methadone) modalities. Given the known challenges of treating these clients in drug-free settings relative to the documented effectiveness of methadone maintenance, these analyses investigate the availability of various clinical and wraparound services for this population among a US sample of addiction treatment programs with and without methadone maintenance services (N = 763). Face-to-face interviews conducted in 2002-2003 gathered data on the number of opiate-dependent clients treated; organizational characteristics, including size, ownership, accreditation, and staffing; treatment practices, including methadone availability, use of other pharmacotherapies, and levels of care; and services offered, including vouchers, transportation, and other wraparound services. Facilities treating proportionately more opiate-dependent clients were significantly more likely to offer a variety of evidence-based services, regardless of methadone availability. Implications for referral linkages and quality of care are discussed. PMID- 17127546 TI - Modafinil and cocaine interactions. AB - This Phase I trial evaluated the interaction between modafinil steady-state and cocaine. Twelve non-treatment seeking, cocaine dependent volunteers received four sets of randomized blinded infusions of saline, 20 mg IV cocaine, and 40 mg IV cocaine. Modafinil was given open label at 0 mg, 400 mg, or 800 mg. Modafinil combined with IV cocaine did not result in any significant hemodynamic interactions. Modafinil significantly dampened scores on Visual Analog Scale measures as compared to baseline cocaine conditions. No significant alterations in labs occurred. Further outpatient trials of modafinil appear to be warranted. PMID- 17127547 TI - Substance use and psychosocial outcomes following participation in residential laboratory studies of marijuana, methamphetamine and zolpidem. AB - RATIONALE: Non-therapeutic research with drugs of abuse in humans is important for a more comprehensive understanding of substance abuse and for the development of more effective treatments. However, the administration of substances from drug classes with abuse potential to human volunteers raises ethical questions regarding potential risk to study volunteers. OBJECTIVE: The purpose of this study was to assess the psychosocial functioning and reported drug-taking behavior of volunteers before and after participating in a residential laboratory study, during which either marijuana, methamphetamine or zolpidem was administered. METHODS: Twenty-two volunteers were administered Addiction Severity Index (ASI) interviews at intake and approximately six months following their study participation. RESULTS: No significant differences between intake and follow-up assessments were found on any ASI composite or drug/alcohol-taking variable. CONCLUSION: These preliminary data suggest that participation in residential laboratory studies involving the administration of drugs from classes with abuse potential does not alter subsequent psychosocial functioning or reported drug use. PMID- 17127548 TI - From the eyes of the beholder: alcohol expectancies and valuations as predictors of hazardous drinking behaviors among female college students. AB - Research has shown that drinking expectancies are associated with alcohol use among college students; however, the bulk of these studies have focused exclusively on researcher-labeled "positive" or "negative" expectancies rather than on the student's valuation (i.e., rating of desirability) of these expectancies. The present study examined the utility of expectancies and valuations in predicting hazardous alcohol use in a sample of 330 female college students (mean age = 20.0; 18-25). Hierarchical regression analyses revealed that negative expectancies and favorable valuations of negative and positive expectancies were predictive of elevated hazardous use (controlling for age, athletic membership, and peer use). Expectancy valuations accounted for additional variance in the model beyond that of expectancies. The present findings shed light on the utility of expectancies and valuations of expectancies in predicting hazardous alcohol use among female college students. Future research directions and potential implications for prevention efforts are discussed. PMID- 17127549 TI - Psychiatric morbidity and parental substance use disorder. AB - OBJECTIVE: To assess psychiatric morbidity associated with having no, one, or two parents with Substance Use Disorder (SUD), among patients receiving SUD care. DESIGN: Two measures of psychiatric morbidity included (a) current psychopathology and (b) lifetime use of psychiatric treatment. SETTINGS: Alcohol drug treatment programs were located in two university medical centers. SUBJECTS: Four hundred ninety-five voluntary patients aged 18 and older and non-adopted. RESULTS: Parental SUD was directly related to (a) more current psychiatric symptoms, both self-rated and psychiatrist-rated and (b) greater likelihood of ever having ever received psychiatric treatment. Among those who had ever received psychiatric care, the number of psychotropic medications was related to parental SUD. Number of admissions, venues, visits/days, and cost of psychiatric care were not associated with parental SUD. CONCLUSIONS: Extent of parental SUD was related to increased psychiatric morbidity. PMID- 17127550 TI - Cocaine withdrawal symptoms predict medication response in cocaine users. AB - The purpose of this study was to examine the influence of cocaine withdrawal symptoms on addiction severity and treatment outcomes in methadone stabilized cocaine users who participated in pharmacotherapy trials using gamma-aminobutyric acid (GABA) medications. Subjects who fulfilled DSM-IV cocaine withdrawal criteria (n = 45), compared to those who did not (n = 40), showed a greater increase in cocaine free urines in response to pharmacotherapy with GABA medications. Altogether, our results and previous studies support the clinical utility of cocaine withdrawal symptoms in predicting treatment response to medications, such that low withdrawal severity may predict better treatment response to GABA medications, while high withdrawal severity may predict better response to adrenergic blockers. This hypothesis needs to be tested in prospective clinical trials. PMID- 17127551 TI - Concurrent cannabis use during treatment for comorbid ADHD and cocaine dependence: effects on outcome. AB - Cannabis is the most widely used illicit substance in the United States with especially high prevalence of use among those with psychiatric disorders. Few studies have examined the relationship between concurrent cannabis use and treatment outcome among patients receiving treatment for comorbid substance abuse and psychiatric disorders. This study investigated the effects of cannabis use on treatment retention and abstinence from cocaine among cocaine dependent patients with Attention Deficit Hyperactivity Disorder (ADHD). Cocaine dependent patients diagnosed with current ADHD (DSM-IV, N = 92) aged 25 to 51 participated in a randomized clinical trial of methylphenidate for treatment of ADHD and cocaine dependence in an outpatient setting. The majority of patients (69%) used cannabis during treatment. Results suggest that moderate/intermittent cannabis users had greater retention rates compared to abstainers and consistent users (p = .02). This study is the first to examine concurrent cannabis use in cocaine dependent patients diagnosed with ADHD. PMID- 17127552 TI - Correlates of condom use and reasons for condom non-use among drug users. AB - Two hundred and seventy-seven drug using adults were interviewed regarding details of their most recent sexual encounter. Demographic, attitudinal, and context variables were associated with condom use and non-use. Greater perceived risk of STDs/HIV and positive attitudes toward condoms' effect on sexual pleasure were associated with greater likelihood of reporting condom use. Common reasons for not using condoms included lower perceived risk of contracting HIV/STDs, negative attitudes toward condoms' effect on pleasure, and lack of condom availability. Tailoring messages to modifiable perceptions of risk and condom attitudes may be useful in reducing sexual risk among drug-using individuals. PMID- 17127553 TI - Psychological correlates of trading sex for money among African American crack cocaine smokers. AB - This article compares demographic characteristics, sexual practices, and psychosocial status among 193 African American female crack cocaine users who currently, previously, or never traded sex for money. Current traders were less likely to have a main sexual partner, more likely to have a casual sexual partner, and more likely to smoke larger quantities of crack. There was a significant trend towards current traders reporting lower self-esteem, greater depression and anxiety, poorer decision-making confidence, more hostility, less social conformity, greater risk taking behaviors, and more problems growing up, compared to previous and never traders. These differences suggest that interventions should address self-esteem, risk-taking practices, depression and anxiety as well as other psychosocial factors. PMID- 17127554 TI - Gender specific associations between types of childhood maltreatment and the onset, escalation and severity of substance use in cocaine dependent adults. AB - We examined associations between types of childhood maltreatment and the onset, escalation, and severity of substance use in cocaine dependent adults. In men (n = 55), emotional abuse was associated with a younger age of first alcohol use and a greater severity of substance abuse. In women (n = 32), sexual abuse, emotional abuse, and overall maltreatment was associated with a younger age of first alcohol use, and emotional abuse, emotional neglect, and overall maltreatment was associated with a greater severity of substance abuse. There was no association between childhood maltreatment and age of nicotine or cocaine use. However, age of first alcohol use predicted age of first cocaine use in both genders. All associations were stronger in women. Findings suggest that early intervention for childhood victims, especially females, may delay or prevent the early onset of alcohol use and reduce the risk for a more severe course of addiction. PMID- 17127555 TI - The impact of 9/11 on patients in New York City's substance abuse treatment programs. AB - OBJECTIVES: This article assesses the impact of the attacks on 9/11 in New York City on drug use, relapse, and mental health from the perspective of drug users and patients in substance abuse treatment programs. METHODS: Structured interviews were conducted with 16 administrators and 75 randomly selected patients at 15 substance abuse treatment programs in New York City from December 2002 to April 2003. RESULTS: Drug use and relapse was a significant issue on and after 9/11. While Post Traumatic Stress Disorder was related to drug use, other preexisting mental health problems were not. Men were more likely to relapse than women; however, women were more emotionally affected by events following 9/11. CONCLUSIONS: The attacks on 9/11 may have contributed to relapse among drug users and exacerbated existing mental health problems among patients with a history of concurrent drug use and mental illness. PMID- 17127556 TI - An evaluation of phencyclidine (PCP) psychosis: a retrospective analysis at a state facility. AB - It has been reported in the literature that phencyclidine (PCP) psychosis recovery may take up to 4-6 weeks. This retrospective review sought to determine whether patients with a new onset of PCP psychosis have a longer hospitalization than those patients with new onset functional psychosis. The PCP arm (N = 20) was found to have a significantly shorter hospitalization than those with a new onset functional psychosis (N = 20)-mean 4.8 days (range 1-9) versus 13.6 days (range 3 41), p < .05. In addition, patients with psychosis related to PCP use were treated more aggressively with conventional antipsychotics than patients with a new onset functional psychosis at this facility. PMID- 17127557 TI - Effect of non-invasive ventilation on survival, quality of life, respiratory function and cognition: a review of the literature. AB - Symptoms of nocturnal hypoventilation may negatively influence the quality of life (QoL) of ALS patients long before respiratory failure ensues. Non-invasive mechanical ventilation (NIV) is considered a treatment option for nocturnal hypoventilation. The primary objective of NIV is improving quality of life (QoL). It may also prolong life by several months. A systematic review of the literature was performed to analyse what is known of the effect of NIV on survival, QoL and other outcome measures. A computerized literature search was performed to identify controlled clinical trials and observational studies of treatment of ALS associated nocturnal hypoventilation from 1985 until May 2005. Twelve studies fulfilled the inclusion criteria. Four studies were retrospective, seven prospective and in one study randomization was used. All studies reported beneficial effects of NIV on all outcome measures. In seven studies NIV was associated with prolonged survival in patients tolerant for NIV, and five studies reported an improved QoL. In conclusion, studies on the use of NIV in ALS differ in study design and endpoint definitions. All studies suggest a beneficial effect on QoL and other outcome measures (Evidence level Class II-III). Well-designed randomized controlled trials comparing the effect on QoL and survival have not been performed. PMID- 17127558 TI - Differential expression of genes in amyotrophic lateral sclerosis revealed by profiling the post mortem cortex. AB - The possible causes of ALS are unknown and multiple biological systems have been implicated. The goal of this study was to use gene expression profiling to evaluate a broad spectrum of systems in ALS. For this study, the medial lip of the human motor cortex and adjacent sensory cortex were collected at autopsy from five ALS patients and three normal individuals. Quantitative filter analysis revealed differential expression of mRNAs normalized to internal standards. A significant difference in expression of 275 genes was found in the ALS motor cortex; of the genes whose expression was changed, 10 were up-regulated and 265 were down-regulated. Six of the up-regulated genes were associated with cell surface activity and two were glutamate receptors; the latter is potentially consistent with the idea of excitotoxicity contributing to neurodegeneration in ALS. Of the down-regulated genes, the largest number were associated with transcription followed by those involved in antioxidant systems, inflammation, regulation of motor neuron function, lipid metabolism, protease inhibition, and protection against apoptosis including vascular endothelial growth factor. There were no significant differences in gene expression patterns between the sensory and motor cortex in the ALS brains. A total of 10% of the genes identified by microarray were chosen from each of the gene groups for validation by quantitative real time PCR (QRT-PCR). In order to increase the reliability of our gene array data, newly acquired motor and sensory cortex of ALS and control cases (n = 4 each) were used for validation. Of these, 86.4% changed in the same direction as determined in the microarrays. The gene profile data reported here are consistent with evidence that the ALS brain is characterized by an environment that is permissive for apoptosis, excitotoxicity and abnormal ubiquitination. This gene array study also suggested that a metal imbalance particularly for zinc could exist in ALS. Finally, given the amount of cellular stress that is thought to be part of the pathogenesis in ALS, there was a notable lack of increase in genes required to mount a protective response. This latter observation provides a conceptual framework in which to consider the possibility that ALS could result from a failure to mount adequate protective responses to physiological insults that, left unchecked, could progress to neurodegeneration. PMID- 17127559 TI - Amyotrophic lateral sclerosis: prognostic indicators of survival. AB - Amyotrophic lateral sclerosis (ALS) has a fatal outcome in about three years, but survival is known to vary considerably, making it difficult to predict disease duration in individual cases. The aim of this study was to investigate possible early prognostic factors of ALS survival. We included 123 probable or definite cases of ALS, with disease onset between 1989 and 1998, and with a follow-up of at least one year. Survival functions were obtained using both the Kaplan-Meier and the actuarial methods. Subgroups, formed on the basis of gender, area of residence, work, and age at and site of onset, were compared using the logrank test and Cox's proportional hazards method (survival functions), and applying the Grizzle, Starmer, Koch (1969), and Koch, Johnson, Tolley (1972) methods (one-year survival probability trends). The survival curves dipped sharply in the first three years, followed by a flattening trend, with 50% of patients dying within 2.5 years, and 89% over seven years. The clinical form with lower limb onset was associated with longer survival than the upper limb onset and bulbar forms (median survival: 39, 27, and 25 months, respectively). Survival was also affected by age at onset (median survival: 34, 27, and 23 months for onset <60, 60-75, and >75 years, respectively), area of residence (median survival: 24 months in mountainous areas, 32 elsewhere), and type of work (median survival: 25 months in agricultural workers, 33.5 in others). Gender did not influence survival, whereas percutaneous endoscopic gastrostomy placement and invasive ventilation did. The estimation of individual ALS survival is important to allow the patient to plan for his future and to make optimal use of medical and community resources. Although age at and site of onset, area of residence, and agricultural work were found to influence survival, there remains an unexplained heterogeneous progression of the disease, suggesting the influence of other, as yet unknown, prognostic factors. The identification of a definite set of prognostic factors may allow physicians to make more reliable survival predictions at diagnosis. PMID- 17127560 TI - Multiple transplants of hNT cells into the spinal cord of SOD1 mouse model of familial amyotrophic lateral sclerosis. AB - hNT cells, derived from a human teratocarcinoma cell line, are versatile neuron like cells that have been studied as possible treatment vehicles for neurodegenerative diseases. Previously, we showed the postponement of motor deficit symptoms in a G93A mouse model of amyotrophic lateral sclerosis (ALS) by transplanting hNT cells into the lumbar spinal cord. In this study, we examined the engraftment of hNT cells at multiple sites within the lumbar spinal cord by morphological analysis of neuritic process development. Results demonstrated that cells implanted at multiple sites established neuritic processes of different lengths independent of the number of cell implants. The hNT fiber outgrowth was a maximum of 0.15-0.3 mm from the transplants and mostly spread within the gray matter; interconnections between implants were not found. Therefore, we suggest that the observed postponement of motor deficit symptoms in G93A mice was not a result of neuritic outgrowth from the implanted hNT cells. PMID- 17127561 TI - Increased incidence of FMO1 gene single nucleotide polymorphisms in sporadic amyotrophic lateral sclerosis. AB - Flavin-containing monooxygenases (FMO) represent a gene family involved in the oxidative metabolism of a variety of xenobiotics, pesticides and drugs. A new function for FMO proteins has been recently uncovered: yeast FMO has been demonstrated to take part in maintaining the redox balance, catalysing the oxidation of reduced glutathione (GSH) to glutathione disulfide (GSSG). The GSSG/GSH balance is an important buffering system for reactive oxygen species and its involvement has been documented in ALS and other neurodegenerative disorders. Human FMO genes present different mutations, which may be related to ethnicity, altered metabolic activity and, in some cases, specific diseases. The human FMO1 gene presents 20 single nucleotide polymorphisms (SNPs) located in coding regions, intronic sequences and untranslated regions. The FMO1 gene has also recently been found underexpressed in spinal cord of ALS patients. Using SSCP and direct sequencing, we studied the allelic and genotypic frequency of two 3'UTR SNPs of the FMO1 gene in sporadic ALS patients compared to a healthy control population. We found a significantly higher frequency of these two polymorphisms, exclusive of the female population, in SALS patients compared to controls (p<0.01), suggesting that specific allelic variants of the FMO1 gene might be associated to susceptibility to develop ALS. PMID- 17127562 TI - Manofluorographic evaluation of swallowing in amyotrophic lateral sclerosis and its relationship with clinical evaluation of swallowing. AB - The aim of this cross-sectional study was, first, to identify swallowing dysfunctions in an ALS population of 40 consecutive patients through combined videofluoroscopy and manometry. Secondly, these objective swallowing data were correlated with the functional feeding status as reported by the patient or family member. Videofluoroscopic evaluation showed dysfunctions in the oral phase of swallowing, pharyngeal initiation and pharyngeal transport. In addition, manometric data revealed low tongue driving forces and pharyngeal contraction amplitudes but normal relaxation of the upper oesophageal sphincter (UES). Aspiration was noted in a not negligible number of 9/40 patients. These objective data were then correlated with the clinical swallowing and feeding status, assessed by means of the ALS Swallowing Severity Scale. Patients receiving scores of 6 or lower on the ALSSSS, report dietary consistency changes but are considered 'safe oral feeders'. Nevertheless, our data revealed that these patients showed significant aspiration during videofluoroscopy. Although not every patient with ALS should be referred routinely for radiographic evaluation of swallowing, our findings suggest referral for a radiological examination as soon as the ALSSSS drops to a score of 6 or lower, to evaluate the presence of (silent) aspiration. PMID- 17127563 TI - Investigation of the therapeutic effects of edaravone, a free radical scavenger, on amyotrophic lateral sclerosis (Phase II study). AB - Amyotrophic lateral sclerosis (ALS) is a rare disease involving selective and progressive degeneration and disappearance of motor neurons. Oxidative stress is believed to contribute to its pathogenesis. We have investigated the efficacy and safety of edaravone, a free radical scavenger previously approved for treatment of acute cerebral infarction, in ALS patients. Within an open trial design, 20 subjects with ALS received either 30 mg (5 subjects) or 60 mg (15 subjects) of edaravone via intravenous drip once per day. Two weeks of administration was followed by a two-week observation period. This four-week cycle was repeated six times. The primary endpoint was the change in the revised ALS functional rating scale (ALSFRS-R) score, while the secondary endpoint was 3-nitrotyrosine (3NT) level in cerebrospinal fluid (CSF). Efficacy was evaluated in the 60 mg group. During the six-month treatment period, the decline in the ALSFRS-R score (2.3+/ 3.6 points) was significantly less than that in the six months prior to edaravone administration (4.7+/-2.1 points); the difference between the two was 2.4+/-3.5 points (Wilcoxon signed rank test, p = 0.039). In almost all patients, CSF 3NT, a marker for oxidative stress, was markedly reduced to almost undetectable levels at the end of the six-month treatment period. Data from the present study suggest that edaravone is safe and may delay the progression of functional motor disturbances by reducing oxidative stress in ALS patients. PMID- 17127564 TI - A pilot trial with clenbuterol in amyotrophic lateral sclerosis. PMID- 17127565 TI - Clinical highlights. PMID- 17127566 TI - ALS Research Group (ALSRG): second meeting, a summary report. PMID- 17127568 TI - A study of semantic treatment of three Chinese anomic patients. AB - A treatment combining semantic feature analysis and semantic priming was carried out on three Cantonese-speaking brain-injured individuals with word-finding difficulties. Two of the participants with mild to moderate semantic impairment demonstrated significant progress on naming performance. Treatment effects also generalised to semantically related and unrelated untrained items. However, only one of these two participants was able to maintain the treatment gain for at least one month after the therapy was completed. The third patient with severe semantic deficits did not benefit from the intervention. The different outcomes of these participants to the same intervention were explained in terms of the nature of the treatment approach, the patients' underlying language deficits, and their level of cognitive abilities. PMID- 17127569 TI - Computerised treatment of anomia in acute aphasia: treatment intensity and training size. AB - In this study we analysed the outcome of computer-assisted therapy (CAT) for anomia on eight acute aphasic patients. Since therapy for anomia generally leads to an item-specific effect, the aim of the present study was to investigate whether it is possible to enhance recovery from anomia by increasing the number of treated items. Two periods of five daily written-naming CAT sessions were compared: In one period the CAT included one set of 48 words (single list) and in the other period a double list of 96 items was treated. Seven out of eight patients improved in naming performance for treated items. Overall gains were superior after practising the double list, despite fewer item repetitions. These results suggest that the size of the effect of therapy for anomia depends more on the number of treated items than on the number of repetitions per item. The integration of these results within the framework of studies on intensity is discussed. PMID- 17127570 TI - Assessment of sensory neglect: a study using moving images. AB - A novel modified passive line-bisection test was performed using a computer display with a moving image. In the test, the subjects were required to give verbal responses instead of limb movements. The test, consisting of two subtasks, left-to-right and right-to-left tasks, was applied to patients with unilateral neglect and controls. The patient group showed an obvious mean deviation in both tasks, and showed greater rightward deviation in the right-to-left task than in the left-to-right task. The control group showed little differences between the two tasks. This paper discusses the results obtained from the two different conditions and their correlations. This method provides useful data for the assessment of unilateral neglect. PMID- 17127571 TI - Cognitive remediation in ADHD: effects of periodic non-contingent alerts on sustained attention to response. AB - Few studies have attempted direct cognitive remediation of attention deficits in attention-deficit hyperactivity disorder (ADHD). The present study investigated the efficacy of periodic non-informative alerting cues for improving sustaining attention in ADHD. This technique is known to improve sustained attention in right frontal injury patients and may be effective in ADHD, given that this disorder has also been linked with right frontal dysfunction. Fifteen children with ADHD and 15 matched controls completed four blocks of a modified version of the Sustained Attention to Response Task (SART). Eight random non-contingent alerts were introduced on two of these blocks as a cue for participants to adopt a more supervisory stance to their performance. While the alerting cues did not alter the total number of commission errors committed by ADHD children over a task block, they did produce a significant short-term reduction in commission errors in the period immediately following an alerting cue. Our data demonstrate that sustained attention performance can be enhanced in children with ADHD using a simple cognitive training strategy. Methods from the field of cognitive rehabilitation may be viably applied to the remediation of attention deficits in ADHD. PMID- 17127572 TI - Action errors and dressing disability after stroke: an ecological approach to neuropsychological assessment and intervention. AB - A combination of detailed observation of dressing behaviour and neuropsychological assessment was used to identify the cognitive barriers to independent dressing in a series of eight stroke patients. For those with right hemisphere damage, dressing was disrupted by visuospatial problems or poor sustained attention. Those with left hemisphere damage and ideomotor apraxia were unable to learn the correct procedure to compensate for hemiparesis when dressing. The value of a therapeutic approach based on these observations was assessed in a single-blind randomised multiple-baseline experiment. A baseline phase of conventional dressing therapy which takes no account of individual patterns of cognitive impairment was contrasted with an approach which was formulated for each case. A permutation test demonstrated that there was a significant treatment effect for the right hemisphere cases but there was no therapy-related improvement for those with left or bilateral damage and apraxia. Observation of a naturalistic but controlled task (dressing with a standard item of clothing) appears to allow greater insight into the impact of specific neuropsychological deficits than has been found for more complex naturalistic tasks. The benefits of this ecological approach over conventional approaches to dressing therapy need to be evaluated further in a randomised clinical trial. PMID- 17127573 TI - Frequency of behavioural problems at one year following traumatic brain injury: correspondence between patient and caregiver reports. AB - A group of 62 adults with significant (i.e., ventilation required for > 24 hours) traumatic brain injury (TBI) were assessed approximately one year following their injury. The people with TBI and their primary caregivers completed the patient and relative/friend versions respectively, of the 20-item Head Injury Behaviour Rating Scale (HIBS). Responses by the patient and caregiver groups were compared for the total number of problems reported and the frequency of specific problem behaviours. The caregiver group reported a greater total number of problem behaviours and a higher frequency for 19 of the 20 specific behaviours. These differences between the patients' and caregivers' reports were statistically significant for seven of the 19 problem behaviours. The majority (86%) of these significant differences were on items from the Behavioural Regulation, rather than the Emotional Regulation, subscale of the HIBS. The implications of these findings for the practice of neuropsychological rehabilitation are presented. PMID- 17127576 TI - Why self-report "Likert" scale data should not be log-transformed. PMID- 17127577 TI - The ecological validity of laboratory cycling: Does body size explain the difference between laboratory- and field-based cycling performance? AB - Previous researchers have identified significant differences between laboratory and road cycling performances. To establish the ecological validity of laboratory time-trial cycling performances, the causes of such differences should be understood. Hence, the purpose of the present study was to quantify differences between laboratory- and road-based time-trial cycling and to establish to what extent body size [mass (m) and height (h)] may help to explain such differences. Twenty-three male competitive, but non-elite, cyclists completed two 25 mile time trials, one in the laboratory using an air-braked ergometer (Kingcycle) and the other outdoors on a local road course over relatively flat terrain. Although laboratory speed was a reasonably strong predictor of road speed (R2 = 69.3%), a significant 4% difference (P < 0.001) in cycling speed was identified (laboratory vs. road speed: 40.4 +/- 3.02 vs. 38.7 +/- 3.55 km x h(-1); mean +/- s). When linear regression was used to predict these differences (Diff) in cycling speeds, the following equation was obtained: Diff (km x h(-1)) = 24.9 - 0.0969 x m - 10.7 x h, R2 = 52.1% and the standard deviation of residuals about the fitted regression line = 1.428 (km . h-1). The difference between road and laboratory cycling speeds (km x h(-1)) was found to be minimal for small individuals (mass = 65 kg and height = 1.738 m) but larger riders would appear to benefit from the fixed resistance in the laboratory compared with the progressively increasing drag due to increased body size that would be experienced in the field. This difference was found to be proportional to the cyclists' body surface area that we speculate might be associated with the cyclists' frontal surface area. PMID- 17127578 TI - Coping in sport: A systematic review. AB - The aim of this paper was to systematically review the literature on coping in sport, examining evidence for both the trait and process perspectives, the types of coping strategies used by athletes, gender differences, age-related differences, and coping effectiveness. A comprehensive literature search of SPORTdiscus, PsychLIT, and PsychINFO in November 2004 yielded 64 studies spanning 16 years (1988 - 2004). The results indicated that athletes use a variety of coping strategies. Forty-six papers supported or adopted the process perspective (Lazarus, 1999; Lazarus & Folkman, 1984). There were also gender and age-related differences. Evidence was found to support three of the different models of coping effectiveness (goodness-of-fit approach, choice of coping strategy, and automacity). Based on this evidence, future research should address some of the methodological and measurement limitations of the sport psychology coping literature. In particular, prospective research designs that minimize the time delay between recall and the stressful experience are required to assess how coping changes over time. More attention to developmental issues to guide the formulation of sport-specific models to enhance our theoretical understanding is also required. Finally, coping effectiveness should be examined both in the short and long term, as a greater understanding of coping effectiveness has the potential to make a significant impact on applied practice. PMID- 17127579 TI - Stressors, social support, and effects upon performance in golf. AB - In this study, we extended the work of Rees and Hardy (2004) by examining the main and stress-buffering effects of social support upon sports performance in a different context, using a different outcome measure, and a specific time-frame. A high-level performance sample of 117 male golfers (mean age 24.8, s = 8.3) completed measures of social support and stressors before competitions. Performance outcome was recorded. Moderated hierarchical regression analyses revealed significant (P < 0.05) main effects for stressors upon performance in 8 of the 11 models tested (R2 = 0.08 - 0.21). Over and above the variance accounted for by stressors, there were significant (P < 0.05) main effects for social support upon performance in all models tested (DeltaR2 = 0.10 - 0.24). In all models, stressors were associated with worse performance, whereas social support was associated with better performance. There were no significant interactions (stress-buffering effects). Main effects for social support upon performance suggest that social support may have aided performance directly, regardless of the level of stress. PMID- 17127580 TI - Olympic lightweight and open-class rowers possess distinctive physical and proportionality characteristics. AB - Rowers competing at the 2000 Olympic Games were measured for 38 anthropometric dimensions. The aim was to identify common physical characteristics that could provide a competitive advantage. The participants included 140 male open-class rowers, 69 female open-class rowers, 50 male lightweight rowers, and 14 female lightweight rowers. Body mass, stature, and sitting height were different (P < 0.01) between the open-class and lightweight rowers, as well as a comparison group of healthy young adults ("non-rowers", 42 males, 71 females), for both sexes. After scaling for stature, the open-class rowers remained proportionally heavier than the non-rowers, with greater proportional chest, waist, and thigh dimensions (P < 0.01). Rowers across all categories possessed a proportionally smaller hip girth than the non-rowers (P < 0.01), which suggested the equipment places some constraints on this dimension. Top-ranked male open-class rowers were significantly taller and heavier and had a greater sitting height (P < 0.01) than their lower-ranked counterparts. They were also more muscular in the upper body, as indicated by a larger relaxed arm girth and forearm girth (P < 0.01). For the male lightweight rowers, only proportional thigh length was greater in the best competitors (P < 0.01). In the female open-class rowers, skinfold thicknesses were lower in the more highly placed competitors (P < 0.01). In conclusion, the rowers in this sample demonstrated distinctive physical characteristics that distinguish them from non-rowers and other sports performers. PMID- 17127581 TI - Changes in running economy following downhill running. AB - In this study, we examined the time course of changes in running economy following a 30-min downhill (-15%) run at 70% peak aerobic power (VO2peak). Ten young men performed level running at 65, 75, and 85% VO2peak (5 min for each intensity) before, immediately after, and 1 - 5 days after the downhill run, at which times oxygen consumption (VO2), minute ventilation, the respiratory exchange ratio (RER), heart rate, ratings of perceived exertion (RPE), and blood lactate concentration were measured. Stride length, stride frequency, and range of motion of the ankle, knee, and hip joints during the level runs were analysed using high-speed (120-Hz) video images. Downhill running induced reductions (7 - 21%, P < 0.05) in maximal isometric strength of the knee extensors, three- to six fold increases in plasma creatine kinase activity and myoglobin concentration, and muscle soreness for 4 days after the downhill run. Oxygen consumption increased (4 - 7%, P < 0.05) immediately to 3 days after downhill running. There were also increases (P < 0.05) in heart rate, minute ventilation, RER, RPE, blood lactate concentration, and stride frequency, as well as reductions in stride length and range of motion of the ankle and knee. The results suggest that changes in running form and compromised muscle function due to muscle damage contribute to the reduction in running economy for 3 days after downhill running. PMID- 17127582 TI - Decision-making skills and deliberate practice in elite association football referees. AB - We examined sport expertise as a function of role. In study 1, referees were better than players in a video-based decision-making task. This provides evidence that there are role-specific skills within one domain or sport. In study 2, we examined the training activities that could be influential in the development of skills in sports officials. Elite association football (soccer) referees retrospectively reported time spent in and perceptions of training activities for three periods: their first year of formal refereeing, 1998 (before formal training programmes were available), and the current year (2003). This allowed us to examine an area of skill with a limited culture of practice, where performance simulations with direct feedback are usually not feasible. The results showed that referees specialize early and, as they develop, they engage in greater volumes and types of training. Competitive match refereeing is considered a relevant activity for skill acquisition that does not fit Ericsson and colleagues' (1993) original definition of deliberate practice. Our findings indicate that actual performance is a significant activity for skill acquisition and refinement. PMID- 17127583 TI - Sprint starts and the minimum auditory reaction time. AB - The simple auditory reaction time is one of the fastest reaction times and is thought to be rarely less than 100 ms. The current false start criterion in a sprint used by the International Association of Athletics Federations is based on this assumed auditory reaction time of 100 ms. However, there is evidence, both anecdotal and from reflex research, that simple auditory reaction times of less than 100 ms can be achieved. Reaction time in nine athletes performing sprint starts in four conditions was measured using starting blocks instrumented with piezoelectric force transducers in each footplate that were synchronized with the starting signal. Only three conditions were used to calculate reaction times. The pre-motor and pseudo-motor time for two athletes were also measured across 13 muscles using surface electromyography (EMG) synchronized with the rest of the system. Five of the athletes had mean reaction times of less than 100 ms in at least one condition and 20% of all starts in the first two conditions had a reaction time of less than 100 ms. The results demonstrate that the neuromuscular physiological component of simple auditory reaction times can be under 85 ms and that EMG latencies can be under 60 ms. PMID- 17127584 TI - Comparison of maximal anaerobic running tests on a treadmill and track. AB - To develop a track version of the maximal anaerobic running test, 10 sprint runners and 12 distance runners performed the test on a treadmill and on a track. The treadmill test consisted of incremental 20-s runs with a 100-s recovery between the runs. On the track, 20-s runs were replaced by 150-m runs. To determine the blood lactate versus running velocity curve, fingertip blood samples were taken for analysis of blood lactate concentration at rest and after each run. For both the treadmill and track protocols, maximal running velocity (v max), the velocities associated with blood lactate concentrations of 10 mmol x l 1 (v10 mM) and 5 mmol x l(-1) (v5 mM), and the peak blood lactate concentration were determined. The results of both protocols were compared with the seasonal best 400-m runs for the sprint runners and seasonal best 1000-m time-trials for the distance runners. Maximal running velocity was significantly higher on the track (7.57 +/- 0.79 m x s(-1)) than on the treadmill (7.13 +/- 0.75 m x s(-1)), and sprint runners had significantly higher vmax, v10 mM, and peak blood lactate concentration than distance runners (P < 0.05). The Pearson product--moment correlation coefficients between the variables for the track and treadmill protocols were 0.96 (v max), 0.82 (v10 mM), 0.70 (v5 mM), and 0.78 (peak blood lactate concentration) (P < 0.05). In sprint runners, the velocity of the seasonal best 400-m run correlated positively with vmax in the treadmill (r = 0.90, P < 0.001) and track protocols (r = 0.92, P < 0.001). In distance runners, a positive correlation was observed between the velocity of the 1000-m time-trial and vmax in the treadmill (r = 0.70, P < 0.01) and track protocols (r = 0.63, P < 0.05). It is apparent that the results from the track protocol are related to, and in agreement with, the results of the treadmill protocol. In conclusion, the track version of the maximal anaerobic running test is a valid means of measuring different determinants of sprint running performance. PMID- 17127585 TI - Changes in the perception of action possibilities while climbing to fatigue on a climbing wall. AB - In two experiments we examined changes in the perception of action possibilities as a function of exertion. In Experiment 1, participants repeatedly climbed on a climbing wall in a series of trials that progressively increased in number to 10 trials, resulting in increased exertion. Before and during climbing, the participants judged their maximum reaching height and perceived exertion. On a separate day, participants climbed another 10 trials while performing actual maximum reaches. Higher perceived exertion was associated with decreases in perceived maximum reach while the actual reaches did not decrease. However, the perceptual changes occurred early during task execution when the participants were not yet fatigued. When exertion set in, neither perceived nor actual maximum reaching appeared to be affected. In Experiment 2, we included exhaustion trials. The findings replicated the early changes in perception observed in Experiment 1, which may be explained by hands-on experience with the task. Furthermore, while climbing to exhaustion, perceptual judgements largely changed in keeping with changes in the actual maximum reach. Thus, there appeared to be a functional relationship between participants' actual action capabilities, rather than their state of physical fatigue per se, and perceived action possibilities. PMID- 17127586 TI - Whole-body vibration can reduce calciuria induced by high protein intakes and may counteract bone resorption: A preliminary study. AB - Excess protein intake can adversely affect the bone via an increase in calcium excretion, while suitable mechanical loading promotes osteogenesis. We therefore investigated whether vibration exposure could alleviate the bone mineral losses associated with a metabolic acidosis. Ten healthy individuals aged 22 - 29 years (median = 25) underwent three 5-day study periods while monitoring their dietary intake. The study consisted of recording the participants' usual dietary intake for 5 consecutive days. Participants were then randomly divided into two groups, one of which received a protein supplement (2 g x kg(-1) body mass x day(-1); n = 5) and the other whole-body low-magnitude (3.5 g), low-frequency (30 Hz) mechanical vibration (WBV) delivered through a specially designed vibrating plate for 10 min each day (n = 5). Finally, for the third treatment period, all participants consumed the protein supplement added to their normal diet and were exposed to WBV exercise for 10 min per day. Daily urine samples were collected throughout the experimental periods to determine the excretion of calcium, phosphate, titratable acid, urea, and C-telopeptide. As expected, when the participants underwent the high protein intake, there was an increase in urinary excretion rates of calcium (P < 0.001), phosphate (P < 0.003), urea (P < 0.001), titratable acid (P < 0.001), and C-telopeptide (P < 0.05) compared with baseline values. However, high protein intake coupled with vibration stimulation resulted in a significant reduction in urinary calcium (P = 0.006), phosphate excretion (P = 0.021), and C-telopeptide (P < 0.05) compared with protein intake alone, but did not affect titratable acid and urea output. The participants showed no effect of WBV exercise alone on urinary excretion of calcium, phosphate, urea, titratable acid, or C-telopeptide. The results indicate that vibration stimulation can moderate the increase in bone resorption and reduction in bone formation caused by a metabolic acidosis. PMID- 17127587 TI - Kicks from the penalty mark in soccer: the roles of stress, skill, and fatigue for kick outcomes. AB - The soccer "penalty shootout" in the knock-out phase of major international tournaments is one of the most dramatic events in international soccer. The outcome of these kicks is typically attributed to factors such as psychology (e.g. coping with stress), skill (e.g. kicking technique), physiology (e.g. overcoming the fatigue of 120 min play), and chance (e.g. in what direction the goalkeeper moves). The purpose of this study was to use internet game record data to estimate the relative importance of these performance components for the outcome of penalty kicks in international matches. Data were collected from soccer statistics internet sites on all 41 penalty shootouts and 409 kicks taken in the World Cup, European Championships, and Copa America between 1976 and 2004. The results showed that the importance of the kicks (indicative of stress) was negatively related to the outcomes of the kicks, whereas skill and fatigue were less, or not, related to outcome. It was concluded that psychological components are most influential for the outcome of penalty kicks. We recommend that practitioners work with players to reduce the perceived importance of each kick. PMID- 17127588 TI - Effect of expertise on butterfly stroke coordination. AB - The aim of this study was to compare the arm-to-leg coordination in the butterfly stroke of three groups of male swimmers of varying skill (10 elite, 10 non-elite, and 10 young swimmers) at four race paces (400-m, 200-m, 100-m, and 50-m paces). Using qualitative video analysis and a hip velocity-video system (50 Hz), key events of the arm and leg movement cycles were defined and four-point estimates of relative phase were used to estimate the arm-to-leg coordination between the propulsive (pull and push of arms and downward movement of leg undulation) and non-propulsive phases (entry, catch, and recovery of arms and upward movement of leg undulation). With increasing race pace, the velocity, stroke rate, and synchronization between the arm and leg key points also increased, indicating that velocity and stroke rate may operate as control parameters. Finally, these changes led to greater continuity between the propulsive actions, which is favourable for improving the swim velocity, suggesting that coaches and swimmers should monitor arm-to-leg coordination. PMID- 17127589 TI - The weighted walking test as an alternative method of assessing aerobic power. AB - The aim of the present study was to determine maximal oxygen uptake (VO2max) directly during uphill walking exercise and to compare these values with those achieved during running and cycling exercise. Forty untrained students (20 males and 20 females) took part in three exercise tests. The running test was performed on a horizontal treadmill and the speed was gradually increased by 0.3 m . s(-1) every 3 min. The walking test was conducted on a treadmill inclined at 12% (speed of 1.8 m . s(-1)). The load was further increased every 3 min by the addition of a mass of one-twentieth of the body mass of the participant (plastic containers filled with water and added to a backpack carried by the participant). During the bicycle ergometry test, the workload was increased by 20 W every 2 min. All tests were performed until volitional exhaustion. During all tests, oxygen uptake, minute ventilation, tidal volume, respiratory frequency, heart rate, hydrogen ion concentration, base excess, and blood lactate concentration were analysed. The Pearson correlation coefficients between the weighted walking test and the commonly applied running and bicycle ergometry tests indicate a strong association with the new test in evaluating maximal oxygen uptake. The negligible differences in VO2max between the three tests for the male participants (running: 61.0 ml . kg(-1) . min(-1); walking: 60.4 ml . kg(-1) . min(-1); cycling: 60.2 ml . kg(-1) . min(-1)), and the fact that the females achieved better results on the walking test than the cycle ergometer test (running: 45.0 ml . kg(-1) . min(-1); walking: 42.6 ml . kg(-1) . min(-1); cycling: 40.1 ml . kg(-1) . min(-1)), confirm the suitability of the new method for evaluating aerobic power. The weighted walking test could be useful in the assessment of aerobic power in individuals for whom running is not advised or is difficult. In addition, the new test allows for determination of VO2max on small treadmills with a limited speed regulator, such as those found in specialist physiotherapy and fitness centres. PMID- 17127590 TI - Global self-esteem, goal achievement orientations, and self-determined behavioural regulations in a physical education setting. AB - We examined a theoretical model of global self-esteem that incorporated constructs from achievement goal and self-determination theories. The model hypothesized that self-determined or autonomous motives would mediate the influence of achievement goal orientation on global self-esteem. The adapted version of the Behavioural Regulation in Exercise Questionnaire (Mullan et al., 1997), the Perception of Success Questionnaire (Roberts & Balague, 1991), and Rosenberg's (1965) self-esteem scales were administered to 634 high school students aged 11 - 15 years. A structural equation model supported the hypotheses and demonstrated that autonomous motives mediated the effect of goal orientations on global self-esteem. The results suggest that generalized motivational orientations influence self-esteem by affecting autonomous motivation and is consistent with theory that suggests that experiences relating to intrinsic motivation are the mechanism by which global motivational orientations are translated into adaptive outcomes like self-esteem. The findings suggest that physical activity interventions that target autonomous motives in physical activity contexts are likely to enhance young people's general self-esteem. PMID- 17127591 TI - Event-related potential evidence for the processing efficiency theory. AB - The purpose of this study was to examine the central tenets of the processing efficiency theory using psychophysiological measures of attention and effort. Twenty-eight participants were divided equally into either a high or low trait anxiety group. They were then required to perform a simulated driving task while responding to one of four target light-emitting diodes. Cortical activity and dual task performance were recorded under two conditions -- baseline and competition -- with cognitive anxiety being elevated in the competitive session by an instructional set. Although driving speed was similar across sessions, a reduction in P3 amplitude to cue onset in the light detection task occurred for both groups during the competitive session, suggesting a reduction in processing efficiency as participants became more state anxious. Our findings provide more comprehensive and mechanistic evidence for processing efficiency theory, and confirm that increases in cognitive anxiety can result in a reduction of processing efficiency with little change in performance effectiveness. PMID- 17127592 TI - Situated analysis of elite trampolinists' problems in competition using retrospective interviews. AB - The aims of this study were to identify and analyse elite athletes' problems in competition. A situated cognition approach placed the emphasis on athletes' actions (i.e. cognitions and behaviours), which were considered to emerge from couplings with selected elements of the context. Fifty-two exercises performed by 10 elite trampolinists were analysed. Field observations, structured interviews, and self-confrontation interviews were conducted and transcribed, and used together with behavioural descriptions derived from video recordings. Performance problems were selected from these reports and from the major infringements of trampoline rules. Qualitative analysis identified the meaningful units of action and their semiotic components for each problem. Four categories of problem were identified and noted to appear either separately or jointly while performing: (a) finding the best moment to begin the performance; (b) finding and maintaining the best mode of involvement to end the performance; (c) recovering normal sensory motor capacity to perform; and (d) solving problems quickly and definitively while performing. The results suggest that the study of action -- situation couplings in sports, as well as of their constantly evolving dynamics, not only reveals elite athletes' psychological activity, but is vital to a deeper understanding of these couplings. PMID- 17127593 TI - The effect of ball-carrying technique and experience on sprinting in rugby union. AB - There are several ways of carrying the ball in rugby union, which could influence the speed at which a player can run. We assessed 52 rugby players (34 males, 18 females) during a maximum sprint over 30 m without the ball, with the ball under one arm, and with the ball in both hands. Timing gates were used to measure time over the initial 10 m and the last 20 m. It has previously been reported (Grant et al., 2003) that running with the ball produces a slower sprinting speed than running without the ball. We hypothesized that the decrease in speed caused by carrying the ball would become less marked with the experience of the player. The male and female players were each divided into two groups: a "beginner" group that consisted of players in their first or second season and an "experienced" group that was composed of players who had played for more than two seasons. A 2 x 3 mixed-model analysis of variance was used to identify differences (P < 0.01) between the beginner and experienced groups in the three sprinting conditions. The times for the males for the first 10 m sprints without the ball, with the ball under one arm, and with the ball in both hands were 1.87 +/- 0.08 s, 1.87 +/ 0.08 s, and 1.91 +/- 0.1 s for the beginners, and 1.87 +/- 0.1, 1.88 +/- 0.1 and 1.88 +/- 0.12 for the more experienced players respectively. The times for the females for the first 10 m without the ball, with the ball under one arm, and with the ball in both hands were 2.13 +/- 0.16 s, 2.19 +/- 0.17 s, and 2.20 +/- 0.16 s for the beginners, and 2.03 +/- 0.12 s, 2.03 +/- 0.09 s, and 2.04 +/- 0.1 s for the more experienced players respectively. For the last 20 m of the 30-m sprint, there were differences between the different sprint conditions (P < 0.001) but no differences that were attributable to experience (P = 0.297). The times for the males over the last 20 m without the ball, with the ball under one arm, and with the ball in both hands were 2.58 +/- 0.19 s, 2.61 +/- 0.12 s, and 2.65 +/- 0.12 s for the beginners, and 2.59 +/- 0.12, 2.62 +/- 0.23, and 2.65 +/- 0.18 s for the more experienced players respectively. The times for the females over the last 20 m without the ball, with the ball under one arm, and with the ball in both hands were 3.25 +/- 0.38 s, 3.35 +/- 0.42 s, and 3.40 +/- 0.46 s for the beginners, and 3.04 +/- 0.32 s, 3.06 +/- 0.22 s, and 3.13 +/- 0.27 s for the more experienced players respectively. No gender-specific differences were detected. The results of this study suggest that practising sprints while carrying a ball benefits the early phase of sprinting while carrying the ball. PMID- 17127594 TI - The influence of squat depth on maximal vertical jump performance. AB - An increase in the period over which a muscle generates force can lead to the generation of greater force and, therefore, for example in jumping, to greater jump height. The aim of this study was to examine the effect of squat depth on maximum vertical jump performance. We hypothesized that jump height would increase with increasing depth of squat due to the greater time available for the generation of muscular force. Ten participants performed jumps from preferred and deep squat positions. A computer model simulated jumps from the different starting postures. The participants showed no difference in jump height in jumps from deep and preferred positions. Simulated jumps produced similar kinematics to the participants' jumps. The optimal squat depth for the simulated jumps was the lowest position the model was able to jump from. Because jumping from a deep squat is rarely practised, it is unlikely that these jumps were optimally coordinated by the participants. Differences in experimental vertical ground reaction force patterns also suggest that jumps from a deep squat are not optimally coordinated. These results suggest there is the potential for athletes to increase jump performance by exploiting a greater range of motion. PMID- 17127595 TI - Contraction force specificity and its relationship to functional performance. AB - Best practice for improving strength and power through resistance strength training has been the subject of much research and subsequent conjecture. Much of the conjecture can be attributed to methodological discrepancies. The type of dynamometry used in testing, the training experience of research participants, the specific technique employed in a lift, and the methods of collection and calculation all impact on the final variables of interest. This review examines contraction force specificity by first addressing the methodological issues surrounding our interpretation of the results. Then we address the kinematics and kinetics associated with single and multiple repetitions in relation to the development of strength, power, and functional performance. This discussion provides the delimitations for analysis of subsequent training studies. Finally, recommendations are formulated with the aim of assisting assessment and training practice as well as providing directions for future research. The results of this review suggest that the enhancements in performance resulting from resistance training are context specific in experienced resistance-trained participants. Thus, specific conditioning could be required to achieve improvements in functional performance in this group. PMID- 17127596 TI - Kinematics, coordination, variability, and biological noise in the prone flutter kick at different levels of a "learn-to-swim" programme. AB - The purpose of this study was to establish the characteristics of the movement patterns common to flutter kicking of skilled swimmers and to determine how the movement patterns of swimmers at different levels of a "learn-to-swim" programme differ from those of skilled swimmers. Also, the nature of the skill afforded the opportunity to investigate learning of a cyclical multi-joint task from a motor control perspective. The underwater motion of nine children representing three levels of a "learn-to-swim" programme and 10 skilled swimmers were video-recorded while performing nine cycles of prone flutter kicking. Kinematics including joint angular motion and coordination of joint actions were calculated. Fourier analysis was applied to determine the frequency composition of the vertical undulations of the hip, knee, and ankle and to calculate the velocity of the body wave travelling caudally from hip to ankle. Fourier analysis also enabled investigation of biological noise, as distinct from variability. The results indicated the desired joint angles and coordination towards which learners could be guided. An index based on the ratio of hip - knee and knee - ankle body wave velocities showed that the inter-joint coordination of most learners was not appropriate for effective flutter kicking. There was strong evidence to suggest that skilled performance in flutter kicking is characterized by sequencing of joint actions to produce a single sinusoidal body wave moving caudally with not decreasing and preferably increasing velocity, low biological noise, and small variability. PMID- 17127599 TI - Implementation of guidelines for the prevention of cardiovascular diseases in Europe: problems and challenges. Is the SCORE prediction system useful, or does it do more harm than good? PMID- 17127600 TI - The importance of total cardiovascular risk assessment in clinical practice. AB - BACKGROUND: The Third Joint European Guidelines on Cardiovascular Disease (CVD) Prevention highlight the importance of total cardiovascular risk prediction in planning preventive strategies. AIM: To review the development of risk prediction systems in Europe and the practicalities of utilizing risk prediction in everyday clinical practice. METHODS: Randomised clinical trials, cohort studies and current guidelines for the management of cardiovascular risk were reviewed. Relevant articles published between 1960 and 2004 were identified on PubMed and the Internet. RESULTS: Several risk factors have been confirmed as major independent predictors of CVD, with their modification reducing cardiovascular risk. Total risk assessment is essential as multiple risk factors confer greater risk than the sum of their components. Framingham Heart Study data have been used as the basis for several risk prediction systems, which have been incorporated into numerous guidelines. However, these data are from a relatively small American cohort and tend to overestimate risk in some European populations. This and other limitations prompted the development of the SCORE (Systematic Coronary Risk Evaluation) risk charts using a much larger dataset from 12 European cohort studies. An electronic version of the charts, HeartScore, is now available to provide practitioners with an interactive risk prediction and management system that can be tailored to individual countries. CONCLUSION: Evidence-based risk assessment systems can facilitate logical, individually tailored risk management and contribute to better patient care. Secular changes in coronary heart disease (CHD) incidence and mortality as well as in risk factors such as obesity and diabetes mean that such systems should be capable of modification and evolution. PMID- 17127601 TI - Auditing secondary prevention of ischaemic heart disease in rural areas of Spain: an opportunity for improvement. AB - AIM: To describe the standard of secondary prevention received by individuals with a history of ischaemic heart disease (IHD) in Spanish rural areas, and the factors associated with low standards of quality. METHODS: Medical audit of patients with a history of IHD, whose data were provided by 72 rural physicians in 12 autonomous regions (1030 cases). Quality criteria were used based on international guidelines. Multivariate analysis was employed to assess the variables associated with poor-quality attention. RESULTS: 30.9% of patients gave target low-density lipoprotein (LDL) readings, and 68.1% reached target blood pressure (BP). Beta-blockers were taken by 48.5%. Twenty-nine per cent of patients had not visited a specialist within the previous year. The fact that patients had visited a specialist within the previous year was associated with their having followed the types of treatments recommended in the guidelines (p < 0.01) and with obtaining target LDL and BP readings (p < 0.05). Patients from the smallest villages had the lowest probability of having LDL controls and also of receiving hypolipidaemic therapy (p<0.01). Those with a past history of isolated angina had lower probability of being treated with antiaggregants than those who had experienced previous acute myocardial infarction (p < 0.01). CONCLUSION: The quality of secondary prevention for these patients shows there is room for improvement. Problems of accessibility exist for some groups, which may be improved with the involvement of rural primary healthcare teams. PMID- 17127602 TI - Quality of oral anticoagulation in patients with atrial fibrillation: a cross sectional study in general practice. AB - OBJECTIVE: To evaluate the quality of management of oral anticoagulation among patients on oral anticoagulation for atrial fibrillation, and to verify the relation between patient performance and the risk of an event due to therapy. METHODS: In a retrospective cross-sectional study involving 66 general practices, international normalized ratio (INR) values obtained over a 6-mo period were analysed. All INR values were determined by a single clinical laboratory, and additional medical information was provided by GPs. RESULTS: 395 patients were included in the study, with a mean age of 74+/-9.6 y. In total, 3111 INR values were obtained. The mean number of tests/month per patient was 2.7+/-4.3. A total of 49 728 d of therapy was evaluated. Fifty-three per cent of the day values were within 0.5 INR units of the target (and 69% within 0.75 INR units of the target). The incidence rate for major bleeding was 4.4/100 patient years (and 2.9/100 patient years for thromboembolic events). There was a significant relation between patient performance and the presence of an event (p=0.017), with an odds ratio of 2.8 (95% CI 1.3-6.3). CONCLUSION: The quality of oral anticoagulation in patients with atrial fibrillation is suboptimal. This is significantly related to an increased risk of haemorrhagic events. PMID- 17127603 TI - Comparing costs of intramuscular and oral vitamin B12 administration in primary care: a cost-minimization analysis. AB - OBJECTIVE: To establish whether savings could be made by changing patients from intramuscular to high doses of oral vitamin B12 in primary care without compromising their wellbeing. METHODS: Cost-minimization analysis from a UK perspective, using secondary data obtained from the literature available and expert opinion. RESULTS: The cost of the resources used to treat patients with vitamin B12 deficiency with intramuscular vitamin B12 was calculated as between 55.99 pounds (83.1 Euro) and 99.99 pounds (148.5 Euro) per year. The cost of treating patients with high doses of oral vitamin B12 during the first year was between 125.55 pounds (186.5 Euro) and 248.55 pounds (369.1 Euro). However, once patients receiving intramuscular treatment had been converted to oral treatment, or in new patients treated orally from the outset, the cost was 35.55 pounds per year (52.8 Euro). One variable, home visits, had a high impact on the calculations. CONCLUSION: Switching patients with vitamin B12 deficiency from intramuscular to high-dose oral therapy and treating patients newly diagnosed with vitamin B12 deficiency with oral vitamin B12 from the outset could save resources in the medium and long term, and in newly diagnosed patients. Savings would come particularly in the form of nursing time. PMID- 17127604 TI - Job satisfaction among general practitioners: a systematic literature review. AB - OBJECTIVE: In recent years, the incidence of being overworked and burnt out has increased among general practitioners (GPs). One of the factors that influences the development of burnout is the job satisfaction that physicians experience. Therefore, we conducted a literature review to answer the question: what factors influence the job satisfaction experienced by GPs? METHODS: We used two methods to retrieve citations. We searched four literature databases for citations from 1990 until July 2006, and we checked the reference lists of relevant articles. The inclusion criteria were: GPs had to be the subjects of the study, the study had to describe empirical research, the study had to focus on job satisfaction, and the number of subjects had to be greater than 30. RESULTS: We found 24 relevant citations. Factors increasing job satisfaction which were mentioned more than twice were: diversity of work, relations and contact with colleagues, and being involved in teaching medical students. Factors decreasing job satisfaction were: low income, too many working hours, administrative burdens, heavy workload, lack of time, and lack of recognition. CONCLUSION: Aspects of job satisfaction concerning the content of the profession seem to increase job satisfaction, and aspects concerning employment conditions seem to decrease job satisfaction. PMID- 17127605 TI - Level of glycaemic and lipid control among diabetic patients in Greek primary care. PMID- 17127606 TI - A glass ceiling within academic general practice in Western Europe: career inhibiting and -facilitating factors for female general practitioners. PMID- 17127607 TI - When "personal skills" are more difficult to learn than "know-how". PMID- 17127612 TI - Exploring the elements of narrative that emerge in the interactions between an 8 year-old child who uses an AAC device and her teacher. AB - Narrative abilities have been identified as a link to successful school achievement and, in particular, to the acquisition of literacy. Children who use AAC may be at risk of impaired narrative facility due to the differences in their language learning experiences, limitation of their AAC systems, and limitations from constrained access to physical and social environments. In this study, the elements of narrative that emerged in the interactions between an 8-year-old child who used an AAC device and her teacher are described. This assessment was achieved through use of the Narrative Assessment Profile (Bliss, McCabe, & Miranda, 1998) in the context of five tasks designed to elicit a spectrum of narrative features. Results indicate that the interactions between the child and her teacher made it difficult to assess whether or not the child had control of certain features of narrative. From a purely structural analysis, most narrative discourse dimensions appeared to be severely compromised and therefore in need of immediate intervention. Discussion includes aspects of narrative intervention and suggested topics for further research. PMID- 17127613 TI - Demonstration of a method for assessing semantic organization and category membership in individuals with autism spectrum disorders and receptive vocabulary limitations. AB - A recognized challenge in the field of augmentative and alternative communication (AAC) is the assessment of the individual skills and preferences of potential users of AAC. Particularly in cognitive assessment, many traditional methods are inappropriate because they require the participant to produce a verbal response and/or involve complex verbal instructions. For individuals with limited verbal forms of language, failure at such tasks is relatively uninstructive, either for revealing their functional intellectual status or for developing effective interventions. This paper presents a demonstration of a method developed to evaluate category structure and, thus, semantic organization in individuals with limited verbal skills concomitant to autism spectrum disorder. This method offers a promising tool for assessing clients for AAC. Further potential uses of this method, both clinically and in research, are discussed. PMID- 17127614 TI - Synthesized speech intelligibility among native speakers and non-native speakers of English. AB - Using synthesized and digitized speech in electronic communication devices may greatly benefit individuals who cannot produce intelligible speech. However, multiple investigations have demonstrated that synthesized speech is not always sufficiently intelligible for its listeners. Listening to synthesized speech may be particularly problematic for listeners for whom English is a second language. We compared native and non-native English-speaking adults' listening accuracy for English sentences in natural voice and synthesized voice conditions. Results indicated a disproportionate disadvantage for the non-native English-speaking group when listening to synthesized speech compared to their native English speaking age peers. There was, however, significant variability in performance within the non-native English group, and this was strongly related to independent measures of English language skill. Specifically, a large portion of the variance in performance on the synthesized speech task was predicted by participants' receptive vocabulary scores. PMID- 17127615 TI - Re-designing scanning to reduce learning demands: the performance of typically developing 2-year-olds. AB - Children with severe motor impairments who cannot use direct selection are typically introduced to scanning as a means of accessing assistive technology. Unfortunately, it is difficult for young children to learn to scan because the design of current scanning techniques does not always make explicit the offer of items from the selection array; furthermore, it does not provide explicit feedback after activation of the switch to select the target item. In the current study, scanning was redesigned to reduce learning demands by making both the offer of items and the feedback upon selection more explicit through the use of animation realized through HTML and speech output with appropriate intonation. Twenty typically developing 2-year-olds without disabilities were randomly assigned to use either traditional scanning or enhanced scanning to select target items from an array of three items. The 2-year-olds did not learn to use traditional scanning across three sessions. Their performance in Session 3 did not differ from that in Session 1; they did not exceed chance levels of accuracy in either session (mean accuracy of 20% for Sessions 1 and 3). In contrast, the children in the enhanced scanning condition demonstrated improvements in accuracy across the three 10-20-min sessions (mean accuracies of 22 and 48% for Sessions 1 and 3, respectively). There were no reliable differences between the children's performances with the two scanning techniques for Session 1; however, by Session 3, the children were more than twice as accurate using the enhanced scanning technique compared to the traditional design. Results suggest that by redesigning scanning, we may be able to reduce some of the learning demands and thereby reduce some of the instructional time required for children to attain mastery. Clinical implications, limitations, and directions for future research and development are discussed. PMID- 17127616 TI - Long-term outcomes for individuals who use augmentative and alternative communication: part I--what is a "good" outcome? AB - Over the past 20 years, there have been many advances in the field of augmentative and alternative communication (AAC). Despite these advances, there are no data on the long-term outcomes of AAC interventions. This study evaluated the long-term outcomes for a group of seven young men (ages 19-23 years) who had used AAC systems for at least 15 years and were part of the first generation to have received AAC services since they were in preschool. Outcomes were measured in the following domains: (a) receptive language; (b) reading comprehension; (c) communicative interaction; (d) linguistic complexity; (e) functional communication; (f) educational and vocational achievement; (g) self determination; and (h) quality of life. The outcomes for the group were diverse, with individual variations across all measures. Evaluation of the data raised many issues surrounding the challenges of outcomes measurement; these are discussed with suggestions for future research. PMID- 17127617 TI - Use of aided language stimulation to improve syntactic performance during a weeklong intervention program. AB - This pilot study assessed the performance of nine children, aged 4;8-14;5, who used augmentative and alternative communication (AAC) systems before and after a weeklong aided language stimulation intervention program. Pre/post test data were analyzed to evaluate the participants' syntactic complexity when using (a) a manual communication board and (b) a dynamic display speech generating device (DD SGD). Results indicate that most participants improved their syntactic performance and that these gains were more pronounced when the participants used a manual communication board as compared with a DD-SGD. There was considerable individual variation in performance. PMID- 17127618 TI - HIV-1, lipid rafts, and antibodies to liposomes: implications for anti-viral neutralizing antibodies. AB - The human immunodeficiency virus type 1 (HIV-1) is an enveloped virus with a lipid bilayer that contains several glycoproteins that are anchored in, or closely associated with, the membrane surface. The envelope proteins have complex interactions with the lipids both on the host cells and on the target cells. The processes of budding from host cells and entry into target cells occur at sites on the plasma membrane, known as lipid rafts, that represent specialized regions that are rich in cholesterol and sphingolipids. Although the envelope glycoproteins are antigenic molecules that potentially might be used for development of broadly neutralizing antibodies in a vaccine to HIV-1, the development of such antibodies that have broad specificities against primary field isolates of virus has been largely thwarted to date by the ability of the envelope proteins to evade the immune system through various mechanisms. In this review, the interactions of HIV-1 with membrane lipids are summarized. Liposomes are commonly used as models for understanding interactions of proteins with membrane lipids; and liposomes have also been used both as carriers for vaccines, and as antigens for induction of antibodies to liposomal lipids. The possibility is proposed that liposomal lipids, or liposome-protein combinations, could be useful as antigens for inducing broadly neutralizing antibodies to HIV-1. PMID- 17127619 TI - Quaternary structure of a SPATE autotransporter protein. AB - The temperature-sensitive hemagglutinin (Tsh) is a representative of the growing subfamily of secreted bacterial virulence factors, known as serine protease autotransporters of the Enterobacteriaceae (SPATEs). Expressed by avian and human pathogenic strains of Escherichia coli Tsh acts as a serine protease and an adhesin to erythrocytes, hemoglobin, and extracellular matrix proteins. Mature Tsh is comprised of a 106-kDa secreted domain (Tshs) and a 33-kDa outer membrane beta-domain (Tshbeta). Based on the size of beta-domains and functional properties of their passenger domains, all SPATEs are considered to be conventional autotransporters. However, it is unsettled if the conventional autotransporters exist as monomers, oligomers, or multimers (e.g., hexamers). To determine the quaternary structure of Tsh in vitro, we purified Tshbeta from the outer membranes and showed that it is natively folded because it is heat modifiable and resistant to protease digestion. Blue-native polyacrylamide gel electrophoresis of Tshbeta indicated that Tshbeta exists as a monomer or a dimer. The cross-linking analysis demonstrated that purified Tshbeta exists as a monomer. The size-exclusion chromatography and cross-linking analyses of purified Tshs also showed that the passenger domain of Tsh is a monomer. Overall, our data indicated that Tsh is a monomeric protein in vitro and support the concept that the SPATE autotransporters exist as monomers rather than as multimers. Implications of our findings on the mechanism of autotransporter secretion across the outer membrane are discussed. PMID- 17127620 TI - Knockout of Arfrp1 leads to disruption of ARF-like1 (ARL1) targeting to the trans Golgi in mouse embryos and HeLa cells. AB - ADP-ribosylation factor related protein 1 (ARFRP1) is a member of the ARF-family of GTPases which operate as molecular switches in the regulation of intracellular protein traffic. Deletion of the mouse Arfrp1 gene leads to embryonic lethality during early gastrulation, suggesting that ARFRP1 is required for cell adhesion related processes. Here we show that ARFRP1 specifically controls targeting of ARL1 and its effector Golgin-245 to the trans-Golgi. GTP-bound ARFRP1 (ARFRP1 Q79L mutant) is associated with Golgi membranes and co-localized with the GTPase ARL1. In contrast, the guanine nucleotide exchange defective ARFRP1 mutant (ARFRP1-T31N) clusters within the cytosol. ARFRP1-T31N or depletion of endogenous ARFRP1 by RNA interference disrupts the Golgi association of ARL1 and of the GRIP domain protein Golgin-245 and alters the distribution of a trans-Golgi network marker, syntaxin 6. In contrast, the targeting of two other Golgi-associated proteins, GM130 and giantin, was unaffected. Furthermore, in Arfrp1-/ - embryos ARL1 dislocated from Golgi membranes whereas it was associated with intracellular membranes in wild-type embryos. These data suggest that lethality of Arfrp1 knockout embryos is due to a specific disruption of protein targeting, e.g., of ARL1 and Golgin-245, to the Golgi. PMID- 17127621 TI - The role of charged residues in the transmembrane helices of monocarboxylate transporter 1 and its ancillary protein basigin in determining plasma membrane expression and catalytic activity. AB - Monocarboxylate transporters MCT1-MCT4 require basigin (CD147) or embigin (gp70), ancillary proteins with a glutamate residue in their single transmembrane (TM) domain, for plasma membrane (PM) expression and activity. Here we use site directed mutagenesis and expression in COS cells or Xenopus oocytes to investigate whether this glutamate (Glu218 in basigin) may charge-pair with a positively charged TM-residue of MCT1. Such residues were predicted using a new molecular model of MCT1 based upon the published structure of the E. coli glycerol-3-phosphate transporter. No evidence was obtained for Arg306 (TM 8) of MCT1 and Glu218 of basigin forming a charge-pair; indeed E218Q-basigin could replace WT-basigin, although E218R-basigin was inactive. No PM expression of R306E-MCT1 or D302R-MCT1 was observed but D302R/R306D-MCT1 reached the PM, as did R306K-MCT1. However, both were catalytically inactive suggesting that Arg306 and Asp302 form a charge-pair in either orientation, but their precise geometry is essential for catalytic activity. Mutation of Arg86 to Glu or Gln within TM3 of MCT1 had no effect on plasma membrane expression or activity of MCT1. However, unlike WT-MCT1, these mutants enabled expression of E218R-basigin at the plasma membrane of COS cells. We propose that TM3 of MCT1 lies alongside the TM of basigin with Arg86 adjacent to Glu218 of basigin. Only when both these residues are positively charged (E218R-basigin with WT-MCT1) is this interaction prevented; all other residue pairings at these positions may be accommodated by charge-pairing or stabilization of unionized residues through hydrogen bonding or local distortion of the helical structure. PMID- 17127622 TI - Molecular dynamics simulations of a bacterial autotransporter: NalP from Neisseria meningitidis. AB - NalP is an autotransporter secretory protein found in the outer membrane of Neisseria meningitidis. The crystal structure of the NalP translocator domain revealed a transmembrane beta-barrel containing a central alpha-helix. The role of this alpha-helix, and of the conformational dynamics of the beta-barrel pore have been studied via atomistic molecular dynamics simulations. Three simulations, each of 10 ns duration, of NalP embedded within a solvated DMPC bilayer were performed. The helix was removed from the barrel interior in one simulation. The conformational stability of the protein is similar to that of other outer membrane proteins, e.g., OmpA, in comparable simulations. The transmembrane beta-barrel is stable even in the absence of the alpha-helix. Removal of the helix results in an influx of water into the pore region, suggesting the helix acts as a 'plug'. Water molecules entering the resultant pore form hydrogen bonds with the barrel lining that compensate for the loss of helix-barrel hydrogen bonds. The dimensions of the pore fluctuate over the course of the simulation revealing it to be flexible, but only wide enough to allow transport of the passenger domain in an unfolded or extended conformation. The simulations help us to understand the role of the central helix in plugging the pore and in maintaining the width of the barrel, and show that the NalP monomer is sufficient for the transport of the passenger domain in an unfolded or extended conformation. PMID- 17127623 TI - Mitochondrial dynamics generate equal distribution but patchwork localization of respiratory Complex I. AB - Highly dynamic mitochondrial morphology is a prerequisite for fusion and fission. Mitochondrial fusion may represent a rescue mechanism for impaired mitochondria by exchanging constituents (proteins, lipids and mitochondrial DNA) and thus maintaining functionality. Here we followed for the first time the dynamics of a protein complex of the respiratory chain during fusion and fission. HeLa cells with differently labelled respiratory Complex I were fused and the dynamics of Complex I were investigated. The mitochondrial proteins spread throughout the whole mitochondrial population within 3 to 6 h after induction of cell fusion. Mitochondria of fused cells displayed a patchy substructure where the differently labelled proteins occupied separate and distinct spaces. This patchy appearance was already--although less pronounced--observed within single mitochondria before fusion, indicating a specific localization of Complex I with restricted diffusion within the inner membrane. These findings substantiate the view of a homogenous mitochondrial population due to constantly rearranging mitochondria, but also indicate the existence of distinct inner mitochondrial sub-compartments for respiratory chain complexes. PMID- 17127624 TI - Electrophysiological characterization of the Cyclophilin D-deleted mitochondrial permeability transition pore. AB - Mitochondria isolated from engineered mice lacking Cyclophilin D (CypD), a component of the Permeability Transition Pore (PTP) complex, can still undergo a Ca2+ -dependent but Cyclosporin A-insensitive permeabilization of the inner membrane. Higher Ca2+ concentrations are required than for wild-type controls. The characteristics of the pore formed in this system were not known, and it has been proposed that they might differ substantially from those of the normal PTP. To test this hypothesis, we have characterized the PTP of isogenic wild-type and CypD- mouse liver mitochondria in patch clamp experiments, which allow biophysical characterization. The pores observed in the two cases, very similar to those of rat liver mitochondria, are indistinguishable according to a number of criteria. The only clear difference is in their sensitivity to Cyclosporin A. CypD is thus shown to be an auxiliary, modulatory component of the "standard" PTP, which forms and has essentially the same properties even in its absence. The observations suggest that Ca2+, CypD, and presumably other inducers and inhibitors act at the level of an activation or assembly process. Activation is separate and upstream of the gating observable on a short or medium-term time scale. Once the pore is activated, its molecular dynamics and biophysical properties may thus be predicted not to depend on the details of the induction process. PMID- 17127625 TI - The weapon potential of human pathogenic fungi. AB - With the exception of Coccidioides spp., human pathogenic fungi are not found among lists of microbes with potential for biological warfare and bioterrorism against humans. However, many human pathogenic fungi are easily obtainable from the environment, are highly dispersible and can cause significant disease after inhalation with relatively low inocula. When the biological and pathogenic attributes of certain human pathogenic fungi are considered using a formula for calculating the relative weapon potential of a microbe it is as apparent that some organisms such as Coccidioides spp. are comparable to other microbes for which there is significant concern. Our analysis suggests that the current indifference to fungi as potential biological weapons against human populations is probably a perception engendered by their non-communicability, lack of history of use or development as biological weapons, and a relatively low incidence of symptomatic disease following natural infection. Awareness of the weapon potential of human pathogenic fungi is an important consideration for greater preparedness against the threat posed by biowarfare and bioterrorism. PMID- 17127626 TI - Fungal diseases: an evolving public health challenge. AB - The emergence of new fungal pathogens and the resurgence of mycotic diseases that had previously been uncommon is a serious and growing public health problem. This review examines the factors involved in the emergence or re-emergence of several mycotic diseases, including coccidioidomycosis and cryptococcosis, over the past two decades. New approaches to prevention and control are also discussed. PMID- 17127627 TI - Isolation of Cryptococcus neoformans var. neoformans serotype D from Eucalypts in South Brazil. AB - Cryptococcus neoformans causes the second most common opportunistic infection in patients with AIDS. In Brazil, 4.5% of the AIDS-related opportunistic infections are caused by C. neoformans and all varieties are recognized as etiological agents of cryptococcosis. This pathogen is a ubiquitous environmental yeast, commonly associated with avian excreta and decaying wood, especially Eucalypt species. The aim of the present study was to search for C. neoformans in Eucalypts and analyze the genotypic diversity of the obtained isolates by RAPD and PCR fingerprinting. All obtained isolates have been C. neoformans var. neoformans, serotype D molecular type VNIV. Serotype D, was isolated from 3 (37.5%) out of 8 cities surveyed in the South Brazilian state Rio Grande do Sul. Nine (9%) out of 99 environmental samples were obtained from Eucalypt species, Eucalyptus calmadulensis and Eucalyptus tereticornis. Molecular analysis using RAPD and PCR-fingerprinting revealed very little genetic diversity in the obtained cryptococcal serotype D isolates. To our knowledge this is the first report of the isolation of serotype D from Eucalyptus trees in Brazil. More studies are required in order to establish the ecological significance of this finding. PMID- 17127628 TI - In vitro effects of fungi isolated from equine hooves on primary human keratinocytes. AB - The effects of two dermatophytes (Microsporum gypseum and Trichophyton mentagrophytes) and four moulds (Scopulariopsis brevicaulis, Alternaria alternata, Geotrichum candidum and Penicillium spp.) on living keratinocyte cultures were examined in vitro using primary human keratinocytes. Rates of apoptosis of infected cells were determined using a colorimetric TUNEL system which detects the characteristic nuclear DNA fragmentation of apoptotic cells. The cytotoxicity of the individual fungi was tested by quantitatively measuring cytosolic enzyme lactate dehydrogenase, released upon cell lysis, in culture supernatants. Additionally, the cell structures within the infected keratinocytes in cultures were examined by scanning electron microscopy. All of the fungi exhibited high cytotoxicity, whereas the development of only the two dermatophytes and the mould Scopulariopsis brevicaulis resulted in distinctly increased apoptosis. Electron microscopy showed that all fungi studied caused similar alterations in the cell structure, with Microsporum gypseum being the most harmful. Increasing loss of cell adhesion as a consequence of a decreasing number of reticulating cell appendices and a reduced cell plasticity were the most evident alterations. PMID- 17127629 TI - Characterization of an extracellular subtilisin protease of Rhizopus microsporus and evidence for its expression during invasive rhinoorbital mycosis. AB - An endoprotease Arp (alkaline Rhizopus protease) was identified and purified to virtual homogeneity from the culture supernatant of an isolate of Rhizopus microsporus var. rhizopodiformis recovered from a non-fatal case of rhinoorbital mucormycosis. N-terminal sequencing of the mature native enzyme was obtained for the first 20 amino acids and revealed high homology to serine proteases of the subtilisin subfamily. Arp migrated in SDS-PAGE with an estimated molecular mass of 33 kDa and had a pI determined to be at pH 8.8. Arp is proteolytically active against various substrates, including elastin, over a broad pH range between 6 and 12 with an optimum at pH 10.5. After invasive mucormycosis, specific antibodies against Arp were detected in stored serum samples taken from the patient from whom the R. microsporus strain of this study had been isolated. Furthermore, in search of factors involved in thrombosis as a typical complication of mucormycosis, a procoagulatory effect of the enzyme has recently been shown. Altogether, these data substantiate the expression of Arp during human rhinoorbital mucormycosis and suggest a role of the enzyme in pathogenesis. PMID- 17127630 TI - Evaluation of pan-fungal PCR assay and Aspergillus antigen detection in the diagnosis of invasive fungal infections in high risk paediatric cancer patients. AB - Profound and prolonged neutropenia following chemotherapy is a major risk factor for systemic fungal infection. As the early diagnosis of invasive fungal infection (IFI) is difficult, these infections are still associated with high morbidity and mortality. Recently, Pan-fungal polymerase chain reaction (PCR) has been a promising aid in rapid, early diagnosis of IFI. During the past few years, increasing numbers of suspected IFIs were encountered at our institution in patients with prolonged neutropenia after intensified immunosuppressive chemotherapy. The aim of this study was to investigate the diagnostic utility of both the aspergillus galactomannan (GM) antigen and the pan-fungal PCR assay in the diagnosis of IFI in high risk febrile neutropenic paediatric cancer patients. During one year period, 91 febrile neutropenic (FN) paediatric cases at high risk for developing IFI while receiving chemotherapy were investigated at National Cancer Institute, Egypt. These patients were subjected to clinical evaluation, chest CT scan, conventional blood cultures for bacterial and fungal pathogens, aspergillus GM antigen detection and PCR assay utilizing pan-fungal primers. Of the 91 FN episodes, 15 were proven IFI; whereas 27 cases were either probable (n=13) or possible IFI (n=14), and 49 were unlikely to be IFI episodes. Based on positive results for proven/probable IFI and compared to culture results, Pan fungal PCR showed sensitivity, specificity, positive and negative predictive values of 75%, 92%, 84% and 87%; respectively. Aspergillus antigen test showed a sensitivity of 79%, specificity of 61%, positive and negative predictive values of 54% and 83%; respectively. A negative PCR in the proven and probable cases was closely related to previous antifungal therapy for a prior history of IFI. In patients at high risk for IFI, neither the sensitivity, nor specificity of the GM test was sufficient. The results of PCR assay was reasonably specific but not very sensitive and had a chance of missing the diagnosis of IFI. The PCR assay seems a promising test for objectively defining IFI, but is not recommended as the only tool for diagnosing IFI. Combining microscopy, culture, and PCR may improve the diagnostic outcome. PMID- 17127631 TI - PCR-based detection of DNA from the human pathogen Blastomyces dermatitidis from natural soil samples. AB - Blastomyces dermatitidis is the dimorphic fungal agent of blastomycosis, a disease that primarily affects humans and dogs. The clinical appearance of this mycosis is well characterized, but there is still little known about its environmental niche, having been isolated from nature only 21 times. We have developed a PCR-based assay to detect B. dermatitidis from soil samples using primers specific to a portion of the promoter region of the BAD1 virulence gene. An internal standard control, pTJV2, was constructed to validate the results from soil samples. Amplification of this control indicated adequate removal of ambient soil inhibitors. The PCR detection limits for the control plasmid and B. dermatitidis genomic DNA were 0.1 and 500 femtograms, respectively. No PCR cross reactivity was observed against bacteria, actinomycetes, and 13 other fungi that were genetically related or found in the same geographic areas. In spiked soil samples, this method was sensitive to 304 copies of pTJV2 DNA and 8,450 live B. dermatitidis yeast cells. Three of eight natural soil samples from a dog kennel near Lexington, KY in which dogs suffered from blastomycosis were positive using the described method, demonstrating its utility in detecting B. dermatitidis in its natural surroundings. PMID- 17127632 TI - Penicillium piceum infection: diagnosis and successful treatment in chronic granulomatous disease. AB - Infections due to Penicillium species other than P.marneffei are rare. We identified a boy with X-linked chronic granulomatous disease (X-CGD) with a pulmonary nodule and adjacent rib osteomyelitis caused by Penicillium piceum. The only sign of infection was an elevated sedimentation rate. P. piceum was isolated by fine needle aspirate and from excised infected tissues. Surgical removal and one year of voriconazole treatment were very well tolerated and led to complete recovery. Microbiological, microscopic and molecular studies support the fungal diagnosis. P. piceum should be considered as a relevant pathogen in immunocompromised patients. PMID- 17127633 TI - B1 cells contribution to susceptibility in experimental paracoccidioidomycosis: immunoglobulin isotypes and repertoire determination. AB - Paracoccidioidomycosis (PCM) is the most prevalent systemic mycosis in Latin America. The experimental murine model has been used to approach the disease, with susceptible and resistant mice strains that reproduce most of the main human immunological features. Since the hypergammaglobulinemia observed in susceptible mice and humans might have an influence on B1 cells, we investigated its role during the experimental infection with Paracoccidiodes brasiliensis. CBA/Nxid mice, deficient in B1 cells, and CBA/Nxid reconstituted with B1 cells isolated from the non-mutant CBA/J strain were infected with 106 yeast forms of P. brasiliensis. At the 8th and 22nd week post infection the DTH response of CBA/Nxid mice was significantly higher after 24 h of P. brasiliensis antigens inoculation and the specific humoral response was reduced, in comparison to CBA/J or recCBA/Nxid. Production of NAbs is a hallmark of the B1 subset. Higher Ig productions to auto antigens such as DNA, MBP and RBC were observed in CBA/J infected mice or recCBA/Nxid. Anti P. brasiliensis IgG2a was produced by CBA/Nxid mice early in infection, while CBA/J or recCBA/Nxid presented increased levels of this isotype only after the 8th week of infection. Furthermore, western blotting analysis showed that CBA/Nxid mice expanded less clones against P. brasiliensis antigens, with weakly detectable anti-gp43 antibodies while CBA/J mice produce IgM anti-gp43 at the 2nd week of infection and IgG anti-gp43 at the 2nd and 8th week. On the other hand, recognition of gp70, a fungal antigen that, as gp43, inhibits macrophage activation was not compromised in B1 deficient mice. These results suggest that B1 cells might have influence in the kinetic of production of protective isotypes of immunoglobulins and their repertoire that could contribute to an early drive towards a Th2 response, affecting the cellular response in susceptible mice during experimental paracoccidiodomycosis. PMID- 17127634 TI - Successful treatment of tongue aspergillosis caused by Aspergillus flavus with liposomal amphotericin B in a child with acute lymphoblastic leukemia. AB - A 5-year-old boy with acute lymphoblastic leukemia and probable pulmonary aspergillosis developed a hard, protuberant, white-yellow and aggressive elongated lesion on the left side of his tongue during a febrile agranulocytopenic episode. Despite the use of broad-spectrum antibiotics and other supportive therapies, the lesion increased to about 2x4 cm in size within two days and became grey-black with an erythemateous, irregular line. Partial excision of the tongue was performed and isolates recovered from the tongue biopsy specimen were identified as Aspergillus flavus. An increase in the systemic dose (7 mg/kg/day) and local intraoral delivery of liposomal amphotericin B was successful in treating the patient and resulted in improved clinical and laboratory findings. Herein, we document the observation of tongue aspergillosis in a leukemic child with probable pulmonary aspergillosis receiving liposomal amphotericin B therapy and the successful treatment of tongue aspergillosis with an increased dose (7mg/kg) of liposomal amphotericin B. To the best of our knowledge, this is the youngest patient with documented intraoral aspergillosis and only the second case of tongue aspergillosis caused by Aspergillus flavus. PMID- 17127635 TI - Successful treatment of invasive Rhizopus infection in a child with thalassemia. AB - The accepted standard for treatment of zygomycetes is amphotericin B and surgical debridement, however recent data suggest that combined treatment modalities may be optimal. Newer anti-fungal agents show success in vitro and in animal models. We present the case of a 10-year-old boy with invasive Rhizopus microsporus var. rhizopodoformis who was successfully treated with a combination of modalities. PMID- 17127636 TI - Disseminated phaeohyphomycosis due to Ochroconis gallopavum in the setting of advanced HIV infection. AB - Disseminated phaeohyphomycosis is a rare and typically fatal infection caused by members of the dematiaceous fungi, and occurs almost universally in the setting of immunocompromise. We herein report a case of systemic phaeohyphomycosis caused by Ochroconis gallopavum in a patient with advanced HIV disease. A possible risk factor for this infection in our patient was heavy marijuana use. This case highlights both the diagnostic and management challenges posed by these infections. To our knowledge, this is the first reported case of disseminated phaeohyphomycosis due to Ochroconis gallopavum in a patient with HIV. PMID- 17127637 TI - Reply to Dr Conti-Diaz. PMID- 17127638 TI - Effects of on-road highway aerosol exposures on autonomic responses in aged, spontaneously hypertensive rats. AB - Epidemiological studies associate ambient particulate pollution with adverse health outcomes in elderly individuals with cardiopulmonary diseases. We hypothesized that freshly generated ultrafine particles (UFP) contribute to these effects, as they are present in high number concentrations on highways and vehicle passengers are exposed directly to them. Aged spontaneously hypertensive rats (9-12 mo) with implanted radiotelemetry devices were exposed to highway aerosol or filtered, gas-denuded (clean) air using an on-road exposure system to examine effects on heart rate (HR) and heart-rate variability (HRV). On the day of exposure, rats were pretreated with low-dose inhaled or injected lipopolysaccharide (LPS) to simulate respiratory tract or systemic inflammation, respectively. Exposures (6 h) in compartmentalized whole-body chambers were performed in an air conditioned compartment of a mobile laboratory on I-90 between Rochester and Buffalo, NY. HRV parameters were calculated from telemetric blood pressure signals and analyzed for the baseline period and for the first 32 h postexposure. The aerosol size (count median diameter = 15-20 nm; geometric standard deviation = 1.4-4.3) and number concentration (1.95-5.62 x 105/cm3) indicated the predominance of UFP. Intraperitoneal LPS significantly affected all of the parameters in a time-dependent manner; response patterns after inhaled or injected LPS pretreatment were similar, but more prolonged and greater in LPS injected rats. A significant effect of highway aerosol was found, irrespective of pretreatment, which resulted in decreased HR in comparison to clean air-exposed rats. This effect was more persistent ( approximately 14 h) in those rats that received ip LPS as compared to saline. The highway aerosol also significantly affected short-term alterations in autonomic control of HR, as evidenced by elevations in normalized high frequency power and decreased vagosympathetic balance. These findings show that environmental exposure concentrations of mixed traffic-related UFP/gas-phase emissions can affect the autonomic nervous system. PMID- 17127639 TI - Cellular mechanisms of mainstream cigarette smoke-induced lung epithelial tight junction permeability changes in vitro. AB - Mainstream cigarette smoke increases the permeability of human airways; however, the mechanism for this increased permeability is poorly defined. Tight junctions between adjacent epithelial cells constitute the physiological barrier to fluid and macromolecules in epithelium. These structures are highly regulated by phosphorylation and their association with the cytoskeleton. The goal of these studies was to identify the signal transduction pathways that regulate smoke induced permeability. Using a physiologically relevant air-liquid interface exposure system, electrically tight monolayers of the human bronchial epithelial cell-line Calu-3 were exposed to fresh, whole mainstream cigarette smoke. This exposure results in a regulated, dose-dependent loss of epithelial barrier function in the lung epithelial monolayers. With cigarette smoke exposure, transepithelial electrical resistance (TER) is decreased and albumin flux is increased, indicating a loss in barrier function to ions and macromolecules, respectively; however, both largely recover in 30 min. Smoke-induced losses of macromolecular barrier function are the result of multicellular junctional reorganization, resulting in increased leak volume rather than leak frequency. Inhibiting Rho kinase (ROCK) significantly reduces the smoke-induced permeability to both ions and macromolecules, while inhibiting protein tyrosine kinases (PTK) only reduces smoke-induced macromolecular permeability. Interestingly, inhibiting myosin light chain kinase (MLCK) exacerbates smoke-induced permeability, indicating that MLCK and ROCK have opposing regulatory roles. Our results demonstrate that the smoke-induced loss of epithelial barrier function in human bronchial epithelium is a regulated process rather than a cytotoxic response. Additionally, our results indicate that activation of PTK and ROCK and inactivation of MLCK contribute to the increased airway permeability caused by mainstream cigarette smoke. PMID- 17127640 TI - 3-week inhalation exposure to cigarette smoke and/or lipopolysaccharide in AKR/J mice. AB - AKR/J mice were exposed to cigarette smoke (CS) and/or lipopolysaccharide (LPS) via inhalation for 3 wk and pulmonary responses were evaluated. The objective was to explore the feasibility of coexposing LPS with cigarette smoke under a subacute exposure, as a surrogate for viral or bacterial insults, that would mimic the pathogenesis of infection-related chronic obstructive pulmonary disease (COPD) exacerbations. The study was the first step in an effort to develop a rodent COPD model in which morphologic lesions of COPD develop in a shorter period of exposure and more closely simulate human COPD. Mice were exposed 6 h/day, 5 days/wk for 3 wk to one of the following: (1) sham control: filtered air; (2) CS: 250 microg/L wet total particulate matter (WTPM) for 5 h/day followed by 1 h/day air; (3) LPS: 0.5 microg/L LPS (055:B5 Escherichia coli; 3,000,000 EU/mg) for the last 1 h/day 2 day/wk (following 5 h/day of filtered air); and (4) CS/LPS: CS 5 h/day followed by air or LPS (2 days/wk) for 1 h/day. After the last exposure, animals were necropsied and subjected to bronchoalveolar lavage (BAL) or histopathology. The BAL neutrophil counts were highest in the LPS group, while macrophage counts were higher in the CS/LPS group than other exposed groups. The LPS group displayed the greatest increases in BAL cytokines, while KC (keratinocyte-derived chemokine) and TARC (thymus and activation-regulated chemokine) were highest in the CS group. The CS/LPS group had generally lower cytokine levels relative to the LPS or CS groups, except for the levels of RANTES and G-CSF (granulocyte-colony stimulating factor) comparable to the LPS group. At microscopic examination of lung sections, cellular inflammatory infiltrates were most notable in the CS/LPS group, which had a diffuse, predominantly macrophage infiltrate with fewer neutrophils. The LPS group had predominantly neutrophils in the pulmonary infiltrate and the CS group had a predominantly macrophage infiltrate in alveolar ducts and adjacent alveoli. Apoptotic labeling of lung cells was highest with the CS/LPS group. In summary, the CS/LPS group displayed greater cellular infiltration and apoptotic responses in the lung with an indication of immunosuppressive effects (lower BAL cytokines) than the CS or LPS group, suggesting that the CS/LPS model shows promise to be further explored as an animal model for studying pathogenesis of COPD exacerbations. A longer term study with interim assessments is needed to confirm that the subacute responses observed in the CS/LPS group will result in greater severity of COPD-related pulmonary lesions following prolonged exposures. PMID- 17127641 TI - Horizontal diffusion elutriation: a new size-separation technique for preparation of rodent-respirable fibers for animal testing. AB - Short-and long-term animal experiments are used to examine the toxicology and biopersistence of various types of fibers. In order to ensure an adequate exposure dose for testing, modern experimental protocols specify that the exposure aerosol (in an inhalation test) or the fibers (in an intratracheal instillation [IT] test) must contain at least a minimum concentration of long (> 20 mum) rodent-respirable fibers. As produced and handled, most fibers contain a distribution of diameters and lengths, only some of which are both long and rodent-respirable. Therefore, it is necessary to size-separate the fibers to enrich the proportion of long, rodent-respirable fibers in the material to be tested. This article presents a new and relatively simple method for size separation that avoids some of the difficulties associated with other methods. The method, termed horizontal diffusion elutriation (HDE), is illustrated by size separating refractory ceramic fiber (RCF) and four polycrystalline alumina (PCA) fibers. PMID- 17127642 TI - Changes in blood manganese concentration and MRI t1 relaxation time during 180 days of stainless steel welding-fume exposure in cynomolgus monkeys. AB - Welders are at risk of being exposed to high concentrations of welding fumes and developing pneumoconiosis or other welding-fume exposure-related diseases. Among such diseases, manganism resulting from welding-fume exposure remains a controversial issue, as although the movement of manganese into specific brain regions has been established, the similar movement of manganese presented with other metals, such as welding fumes, has not been clearly demonstrated as being similar to that of manganese alone. Meanwhile, the competition between Mn and iron for iron transporters, such as transferrin and DMT-1, to the brain has also been implicated in the welding-fume exposure. Thus, the increased signal intensities in the basal ganglia, including the globus pallidus and subcortical frontal white matter, based on T1-weighted magnetic resonances in welders, require further examination as regards the correspondence with an increased manganese concentration. Accordingly, to investigate the movement of manganese after welding-fume exposure, 6 cynomolgus monkeys were acclimated for 1 mo and assigned to 3 dose groups: unexposed, low dose of (total suspended particulate [TSP] 31 mg/m3, 0.9 mg/m3 of Mn), and high dose of total suspended particulate (62 mg/m3 TSP, 1.95 mg/m3 of Mn). The primates were exposed to manual metal-arc stainless steel (MMA-SS) welding fumes for 2 h/day in an inhalation chamber system equipped with an automatic fume generator for 6 mo. Magnetic resonance imaging (MRI) studies of the basal ganglia were conducted before the initiation of exposure and thereafter every month. During the exposure, the blood chemistry was monitored every 2 wk and the concentrations of metal components in the blood were measured every 2 wk and compared with ambient manganese concentrations. The manganese concentrations in the blood did not show any significant increase until after 2 mo of exposure, and then reached a plateau after 90 days of exposure, showing that an exposure period of at least 60 days was required to build up the blood Mn concentration. Furthermore, as the blood Mn concentration continued to build, a continued decrease in the MRI T1 relaxation time in the basal ganglia was also detected. These data suggested that prolonged inhalation of welding fumes induces a high MRI T1 signal intensity with an elevation of the blood manganese level. The presence of a certain amount of iron or other metals, such as Cr and Ni, in the inhaled welding fumes via inhalation was not found to have a significant effect on the uptake of Mn into the brain or the induction of a high MRI T1 signal intensity. PMID- 17127643 TI - Oxidative stress associated with indoor air pollution and sick building syndrome related symptoms among office workers in Taiwan. AB - This study investigated whether sick building syndrome (SBS) complaints and indoor air pollution for office workers are associated with oxidative stress indicated by urinary 8-hydroxydeoxyguanosine (8-OHdG). With informed consent, 389 employees in 87 government offices of 8 high-rise buildings in Taipei city completed self-reported questionnaires on SBS complaints at work in the past month. Urinary 8-OHdG was determined for each study participant and on-site air pollutants were measured for each office in both indoor and outdoor air. The results showed that urinary 8-OHdG had significant associations with volatile organic compounds and carbon dioxide levels in offices, and with urinary cotinine levels. The mean urinary 8-OHdG level was also significantly higher in participants with SBS symptoms than in those without such complaints (6.16 vs. 5.45 mug/g creatinine, p = .047). The mean 8-OHdG increased as the number of SBS symptoms increased. The multivariate logistic regression analyses showed that the adjusted odds ratios (OR) in relation to micrograms per gram creatinine increase in 8-OHdG were statistically significant for eye dryness (1.12), upper respiratory syndrome (1.17) with particularly nose itching (1.25), sneezing (1.51), dry throat (1.21), skin dryness (1.31), and dizziness (1.19). This study indicates that the 8-OHdG level was significantly associated with SBS complaints after controlling for air pollution and smoking. Whether the 8-OHdG can be used as an effective predictor for SBS symptoms deserves further study. PMID- 17127644 TI - Woodsmoke health effects: a review. AB - The sentiment that woodsmoke, being a natural substance, must be benign to humans is still sometimes heard. It is now well established, however, that wood-burning stoves and fireplaces as well as wildland and agricultural fires emit significant quantities of known health-damaging pollutants, including several carcinogenic compounds. Two of the principal gaseous pollutants in woodsmoke, CO and NOx, add to the atmospheric levels of these regulated gases emitted by other combustion sources. Health impacts of exposures to these gases and some of the other woodsmoke constituents (e.g., benzene) are well characterized in thousands of publications. As these gases are indistinguishable no matter where they come from, there is no urgent need to examine their particular health implications in woodsmoke. With this as the backdrop, this review approaches the issue of why woodsmoke may be a special case requiring separate health evaluation through two questions. The first question we address is whether woodsmoke should be regulated and/or managed separately, even though some of its separate constituents are already regulated in many jurisdictions. The second question we address is whether woodsmoke particles pose different levels of risk than other ambient particles of similar size. To address these two key questions, we examine several topics: the chemical and physical nature of woodsmoke; the exposures and epidemiology of smoke from wildland fires and agricultural burning, and related controlled human laboratory exposures to biomass smoke; the epidemiology of outdoor and indoor woodsmoke exposures from residential woodburning in developed countries; and the toxicology of woodsmoke, based on animal exposures and laboratory tests. In addition, a short summary of the exposures and health effects of biomass smoke in developing countries is provided as an additional line of evidence. In the concluding section, we return to the two key issues above to summarize (1) what is currently known about the health effects of inhaled woodsmoke at exposure levels experienced in developed countries, and (2) whether there exists sufficient reason to believe that woodsmoke particles are sufficiently different to warrant separate treatment from other regulated particles. In addition, we provide recommendations for additional woodsmoke research. PMID- 17127647 TI - Towards automatic skill evaluation: detection and segmentation of robot-assisted surgical motions. AB - This paper reports our progress in developing techniques for "parsing" raw motion data from a simple surgical task into a labeled sequence of surgical gestures. The ability to automatically detect and segment surgical motion can be useful in evaluating surgical skill, providing surgical training feedback, or documenting essential aspects of a procedure. If processed online, the information can be used to provide context-specific information or motion enhancements to the surgeon. However, in every case, the key step is to relate recorded motion data to a model of the procedure being performed. Robotic surgical systems such as the da Vinci system from Intuitive Surgical provide a rich source of motion and video data from surgical procedures. The application programming interface (API) of the da Vinci outputs 192 kinematics values at 10 Hz. Through a series of feature processing steps, tailored to this task, the highly redundant features are projected to a compact and discriminative space. The resulting classifier is simple and effective.Cross-validation experiments show that the proposed approach can achieve accuracies higher than 90% when segmenting gestures in a 4-throw suturing task, for both expert and intermediate surgeons. These preliminary results suggest that gesture-specific features can be extracted to provide highly accurate surgical skill evaluation. PMID- 17127648 TI - Visualization and navigation system development and application for stereotactic deep-brain neurosurgeries. AB - We present the development of a visualization and navigation system and its application in pre-operative planning and intra-operative guidance of stereotactic deep-brain neurosurgical procedures for the treatment of Parkinson's disease, chronic pain, and essential tremor. This system incorporates a variety of standardized functional and anatomical information, and is capable of non rigid registration, interactive manipulation, and processing of clinical image data. The integration of a digitized and segmented brain atlas, an electrophysiological database, and collections of final surgical targets from previous patients facilitates the delineation of surgical targets and surrounding structures, as well as functional borders. We conducted studies to compare the surgical target locations identified by an experienced stereotactic neurosurgeon using multiple electrophysiological exploratory trajectories with those located by a non-expert using this system on 70 thalamotomy, pallidotomy, thalamic deep brain stimulation (DBS), and subthalamic nucleus (STN) DBS procedures. The average displacement between the surgical target locations in both groups was 1.95 +/- 0.86 mm, 1.83 +/- 1.07 mm, 1.88 +/- 0.89 mm and 1.61 +/- 0.67 mm for each category of surgeries, respectively, indicating the potential value of our system in stereotactic deep-brain neurosurgical procedures, and demonstrating its capability for accurate surgical target initiation. PMID- 17127649 TI - Surgical navigation display system using volume rendering of intraoperatively scanned CT images. AB - As operative procedures become more complicated, simply increasing the number of devices will not facilitate such operations. It is necessary to consider the ergonomics of the operating environment, especially with regard to the provision of navigation data, the prevention of technical difficulties, and the comfort of the operating room staff. We have designed and created a data-fusion interface that enables volumetric Maximum Intensity Projection (MIP) image navigation using intra-operative mobile 3D-CT data in the OR. The 3D volumetric data reflecting a patient's inner structure is directly displayed on the monitor through video images of the surgical field using a 3D optical tracking system, a ceiling mounted articulating monitor, and a small-size video camera mounted at the back of the monitor. The system performance and accuracy was validated experimentally. This system provides a novel interface for a surgeon with volume rendering of intra-operatively scanned CT images, as opposed to preoperative images. PMID- 17127650 TI - Bone enhancement filtering: application to sinus bone segmentation and simulation of pituitary surgery. AB - The simulation of pituitary gland surgery requires a precise classification of soft tissues, vessels and bones. Bone structures tend to be thin and have diffuse edges in CT data, and thus the common method of thresholding can produce incomplete segmentations. In this paper, we present a novel multi-scale sheet enhancement measure and apply it to paranasal sinus bone segmentation. The measure uses local shape information obtained from an eigenvalue decomposition of the Hessian matrix. It attains a maximum in the middle of a sheet, and also provides local estimates of its width and orientation. These estimates are used to create a vector field orthogonal to bone boundaries, so that a flux maximizing flow algorithm can be applied to recover them. Hence, the sheetness measure has the essential properties to be incorporated into the computation of anatomical models for the simulation of pituitary surgery, enabling it to better account for the presence of sinus bones. We validate the approach quantitatively on synthetic examples, and provide comparisons with existing segmentation techniques on paranasal sinus CT data. PMID- 17127651 TI - Gaze-contingent control for minimally invasive robotic surgery. AB - OBJECTIVE: Recovering tissue depth and deformation during robotically assisted minimally invasive procedures is an important step towards motion compensation, stabilization and co-registration with preoperative data. This work demonstrates that eye gaze derived from binocular eye tracking can be effectively used to recover 3D motion and deformation of the soft tissue. METHODS: A binocular eye tracking device was integrated into the stereoscopic surgical console. After calibration, the 3D fixation point of the participating subjects could be accurately resolved in real time. A CT-scanned phantom heart model was used to demonstrate the accuracy of gaze-contingent depth extraction and motion stabilization of the soft tissue. The dynamic response of the oculomotor system was assessed with the proposed framework by using autoregressive modeling techniques. In vivo data were also used to perform gaze-contingent decoupling of cardiac and respiratory motion. RESULTS: Depth reconstruction, deformation tracking, and motion stabilization of the soft tissue were possible with binocular eye tracking. The dynamic response of the oculomotor system was able to cope with frequencies likely to occur under most routine minimally invasive surgical operations. CONCLUSION: The proposed framework presents a novel approach towards the tight integration of a human and a surgical robot where interaction in response to sensing is required to be under the control of the operating surgeon. PMID- 17127652 TI - Towards robotized beating heart TECABG: assessment of the heart dynamics using high-speed vision. AB - Active robotic filtering is probably the solution for beating heart Totally Endoscopic Coronary Artery Bypass Grafting (TECABG). In this work, we assess the heart motion dynamics by simultaneous use of high-speed imaging of optical markers attached to the heart, ECG signals and ventilator airflow acquisitions. Our goal is to assess the heart motions (shape, velocity, acceleration) in order to be able to make more accurate specifications for a novel, dedicated robot that could follow these motions in real time. Furthermore, using two additional inputs (ECG and airflow), we propose a novel robust prediction algorithm that could be used with a predictive control algorithm to improve the tracking accuracy. PMID- 17127653 TI - A new labdane type diterpenoid from Trollius ledebouri. AB - A new labdane type diterpenoid, (8alpha,13R)-epoxy-14-labden-6beta,7beta-diol-7 beta-D-(4'-acetyl) fucopyranoside, named ledebourene 1, was isolated from the flowers of Trollius ledebouri. The absolute stereostructure was elucidated on the basis of the spectroscopic analysis. PMID- 17127654 TI - A new tetracyclic triterpenoid from the leaves of Azadirachta indica. AB - A new tetracyclic triterpenoid zeeshanol [25,26,27-trinor-apotirucalla-(apoeupha) 6alpha-, 21-dihydroxy, 7alpha-acetoxy, 1,14,22-tri-en-3, 16-dione] (1) along with a known constituent desfurano-6alpha-hydroxyazadiradione (2) have been isolated from the methanolic extract of the leaves of Azadirachta indica. The structure and the relative configurations of 1 were determined by the spectroscopic method (1H- and 13C-NMR, IR, and MS) and 2D-NMR experiments. PMID- 17127655 TI - A new cycloartane glycoside from Camptosorus sibiricus Rupr. AB - A new cycloartane glycoside was isolated from the dried aerial parts of Camptosorus sibiricus Rupr. By means of chemical and spectroscopic methods (IR, 1D and 2D NMR, ESI-MS), the structure was established as 3beta, 7beta, 24beta, 25, 30-pentahydroxycycloartane-3-O-beta-D-glucopyranoside-24-O-beta-D glucopyranosyl(1 --> 2)-D-glucopyranoside (1). PMID- 17127656 TI - (+)-Apollineanin: a new flavanone from Tephrosia apollinea. AB - Chemical investigation of the leaves of Tephrosia apollinea has yielded a new flavanone named (+)-apollineanin, together with two known flavones: (-) semiglabrin and (-)-semiglabrinol. The structure of the new compound was determined on the basis of mass, 1D and 2D NMR spectroscopies including NOE difference spectroscopy. The absolute stereochemistry of (+)-apollineanin was determined by Mosher ester methodology and from CD data. PMID- 17127657 TI - Oxidation studies on Andrographolide. AB - Jones oxidation of Andrographolide (1), gave mixture of three products (3 dehydroandrographolide (5), 3,19-bis dehydroandrographolide (6) and 19 dehydroandrographolide (7). Tritylation of andrographolide at C19-OH resulted to products 8 and diene 9, which can be converted to its acetate 10 and oxidation product 11. PMID- 17127658 TI - A new benzophenone from the stem bark of Garcinia benthami. AB - From the stem bark of Garcinia benthami a new benzophenone, named Salimbenzophenone, was isolated. The structure was determined by means of spectroscopic analysis. PMID- 17127659 TI - A new amide from Thyrocarpus glochidiatus. AB - A new amide, 4-hydroxy-N-[4-[3-(4-hydroxy-phenyl)-E-acryloylamino]-butyl] benzamide, trivially named as glochidiatusamide (1), along with 5 known compounds, beta-sitosterol, daucosterol, coffee acid, ethyl caffeate, and kaempferol-3-O-beta-D-glucoside, were isolated from the ethanol extract of Thyrocarpus glochidiatus Maxim. Their structures were elucidated by the spectral and chemical evidence. PMID- 17127660 TI - Xanthones from Garcinia mangostana (Guttiferae). AB - Studies on the stem of Garcinia mangostana have led to the isolation of one new xanthone mangosharin (1) (2,6-dihydroxy-8-methoxy-5-(3-methylbut-2-enyl) xanthone) and six other prenylated xanthones, alpha-mangostin (2), beta-mangostin (3), garcinone D (4), 1,6-dihydroxy-3,7-dimethoxy-2-(3-methylbut-2-enyl)-xanthone (5), mangostanol (6) and 5,9-dihydroxy-8- methoxy-2,2-dimethyl-7-(3-methylbut-2 enyl)-2H,6H-pyrano-[3,2-b]-xanthene-6-one (7). The structures of these compounds were determined by spectroscopic methods such as 1H NMR, 13C NMR, mass spectrometry (MS) and by comparison with previous studies. All the crude extracts when screened for their larvicidal activities indicated very good toxicity against the larvae of Aedes aegypti. This article reports the isolation and identification of the above compounds as well as bioassay data for the crude extracts. These bioassay data have not been reported before. PMID- 17127661 TI - Phytochemical analysis of the essential oil of Achillea millefolium L. from various European Countries. AB - Variations in the essential oil composition of Achillea millefolium L. growing in Estonia and in other European countries, were determined. The oils were obtained in yields of 0.9-9.5 mL kg-1. A total of 102 components were identified. The quantitatively most important components of yarrow were sabinene, beta-pinene, 1,8-cineole, artemisia ketone, linalool, alpha-thujone, beta-thujone, camphor, borneol, fenchyl acetate, bornyl acetate, (E)-beta-caryophyllene, germacrene D, caryophyllene oxide, beta-bisabolol, delta-cadinol, chamazulene etc. Samples from Estonia contained high amounts of monoterpenes and chamazulene. High amounts of monoterpenes and chamazulene were also found in samples from Hungary, Greek, Moldavia, Latvia, Lithuania and Germany. The oils from France, Belgium, Russia, Armenia, Spain and Italy were rich in oxygenated monoterpenes and contained a little amount of chamazulene. The drugs from Greece, Estonia, Moldavia and Scotland were rich in sesquiterpenes. The Millefolii herba grown in Estonia conforms to the European Pharmacopoeia (EP) standards in the aspect of the essential oil contents. PMID- 17127662 TI - Chemical composition and in vitro antifungal and antioxidant activity of the essential oil and methanolic extract of Teucrium sauvagei Le Houerou. AB - The chemical composition and the in vitro antifungal and antioxidant activity of the essential oil and the methanolic leaf extracts of Teucrium sauvagei Le Houerou, an endemic medicinal plant growing in Tunisia, have been studied. More than 35 constituents having an abundance >or=0.2% were identified in the oil. beta-Eudesmol, T-cadinol, alpha-thujene, gamma-cadinene, and sabinene were the prevalent constituents. Results of the antifungal activity tests indicated that the methanolic extract inhibited the in vitro growth of seven dermatophytes, whereas the essential oil showed average inhibition against only three dermatophytes. In vitro antioxidant properties of the essential oil and the methanolic extract were determined by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid)) assays and compared to those of the synthetic antioxidant Trolox. Due to their antifungal and antioxidant properties, the essential oil and the methanolic extract of T. sauvagei may be of use as natural preservative ingredients in food and/or pharmaceutical industries. PMID- 17127663 TI - Two new naturally occurring optical polyacetylene compounds from Torricellia angulata var intermedia and the determination of their absolute configurations. AB - Four compounds were isolated from Torricellia angulata var intermedia (Harms.) Hu (Torricelliaceae), followed by formation of derivatives, which gave two new polyacetylene compounds 3 and 4 with respectively a chiral carbon center whose absolute configurations were determined unambiguously by the recently developed extended Mosher's method. This is the first time to obtain natural polyacetylenes possessing a chiral hydroxyl group. PMID- 17127664 TI - A new quassinoid, ailantinol H from Ailanthus altissima. AB - A new quassinoid, ailantinol H, was isolated from a methanol extract of the aerial parts of Ailanthus altissima collected in Taiwan. Its structure was established on the basis of 1D and 2D NMR and HREIMS spectroscopic methods. PMID- 17127665 TI - Antimicrobial and antioxidant activities of the methanolic extracts of three Salvia species from Tunisia. AB - This study examines the in vitro antimicrobial and antioxidant activities of the methanolic extracts of three Salvia species from Tunisia: Salvia aegyptiaca L., S. argentea L. and S. verbenaca Ssp. clandestina L. Pugsley. The extracts inhibited the growth of dermatophytes and of bacteria responsible for unpleasant odours to varying degrees; the pathogenic yeasts Candida albicans and Cryptococcus neoformans, the filamentous fungi Aspergillus fumigatus and selected dog otitis bacteria were all resistant to each of the extracts. The extracts were screened for their antioxidant activities using 2,2-diphenyl-1-picrylhydrazyl free radical scavenging and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid) test systems, and gave positive results in both tests. The extracts of S. aegyptiaca were the most active in both tests, followed by those of S. verbenaca, then S. argentea. These results confirm the antimicrobial and antioxidant activities of the genus Salvia and underline the potential of these plants either as natural preservatives or in pharmaceutical applications. PMID- 17127666 TI - Seasonal variation of colchicine content in Colchicum brachyphyllum and Colchicum tunicatum (Colchicaceae). AB - As part of our continuing investigation of Jordanian Colchicum species, (-) colchicine content in C. brachyphyllum Boiss. & Haussk. ex Boiss and Colchicum tunicatum Feinbr (Colchicaceae), growing wild in Jordan, was determined during different growth stages. Using external reference standard, a reverse-phase gradient photo-diode array high performance liquid chromatography (PDA-HPLC) method was adapted. Underground parts in both species and during different growth stages, always showed higher (-)-colchicine content than the above ground parts. In C. brachyphyllum total (-)-colchicine content of underground parts during flowering stage was found to be about 0.15% (wt/wt), while that of aerial parts was only about 0.04% (wt/wt). In C. tunicatum total (-)-colchicine content of underground parts was found to be 0.12% (wt/wt), and 0.13% (wt/wt) during flowering and vegetating stages, respectively, while that of aerial parts was only about 0.04% (wt/wt) and 0.02% (wt/wt), respectively. In C. tunicatum, stems, roots and unripe seeds are the main storages of (-)-colchicine at flowering, vegetating and seeding stages, respectively, while in C. brachyphyllum, corms are the main storage of (-)-colchicine at flowering and seeding stages. PMID- 17127667 TI - Halichondria sulfonic acid, a new HIV-1 inhibitory guanidino-sulfonic acid, and halistanol sulfate isolated from the marine sponge Halichondria rugosa Ridley & Dendy. AB - A new sulfur-containing guanidino derivative, halichondria sulfonic acid (1) showing anti-HIV-1 activity, and halistanol trisulfate (2) with anti-tumor activity have been isolated from the marine sponge Halichondria rugosa Ridley & Dendy collected in the Chinese Southern Sea. The structure of 1 was elucidated by analysis of spectroscopic and crystal data. PMID- 17127668 TI - New anthraquinones from the stem of Morinda citrifolia Linn. AB - Studies on the chemical constituents of the stem of Morinda citrifolia, Linn. have led to the isolation of two new compounds, morindicinone (=2-hydroxy-1,8 dimethoxy-7-methoxymethylanthraquinone, 1) and morindicininone (=4-hydroxymethyl 1,3-dimethoxyanthraquinone, 2), as well as two known constituents, 2 hydroxyanthraquinone (3) and 2-methoxyanthraquinone (4). Their structures were elucidated by spectral analysis including 2D-NMR techniques. PMID- 17127669 TI - Polyporolide, a new acetogenin from the mycelial culture of a Polyporus fungus. AB - A new acetogenin, polyporolide (1), was isolated from the mycelial solid cultures of a Polyporus basidiomycete. Its structure was elucidated as 4-hydroxy-5-(1 hydroxyhexyl)-dihydrofuran-2-one by spectroscopic methods. Its toxicity to brine shrimps was assessed. PMID- 17127670 TI - Unexpected formation of hydroxybiphenylmethane derivatives and some new observations on Labat test. AB - An unexpected synthesis of symmetrical hydroxybiphenylmethanes involving the reaction of 2-hydroxyphenyl benzyl ketones with ethoxymethyl chloride has been observed. Some new interesting observations of Labat test on colorimetric detection of bichalconyloxy, bichalconyl and biflavonylmethanes having oxygenated ortho positions are presented. PMID- 17127672 TI - Road traffic injuries in a two-speed Europe. PMID- 17127673 TI - Stearoyl-CoA desaturase-1 mediates the pro-lipogenic effects of dietary saturated fat. AB - Dietary saturated fats have often been implicated in the promotion of obesity and related disorders. It has been shown recently that saturated fats act through the transcription factor SREBP-1c (sterol regulatory element-binding protein-1c) and its requisite coactivator, peroxisome proliferator-activated receptor-gamma coactivator-1beta (PGC-1beta), to exert their pro-lipogenic effects. We show here that a diet high in the saturated fat stearate induces lipogenic genes in wild type mice, with the induction of the Scd1 (stearoyl-CoA desaturase-1) gene preceding that of other lipogenic genes. However, in Scd1-/- mice, stearate does not induce lipogenesis, and Srebp-1c and Pgc-1beta levels are markedly reduced. Instead, genes of fatty acid oxidation such as Cpt-1 (carnitine palmitoyltransferase-1) as well as Pgc-1alpha are induced. Mitochondrial fatty acid oxidation is increased, and white adipose tissue and hepatic glycogen stores are depleted in stearate-fed Scd1-/- mice. Furthermore, AMP-activated protein kinase is also induced by stearate feeding in Scd1-/- mice. These results indicate that the desaturation of saturated fats such as stearate by SCD is an essential step mediating their induction of lipogenesis. In the absence of SCD1, stearate promotes oxidation, leading to protection from saturated fat-induced obesity. SCD1 thus serves as a molecular switch in the promotion or prevention of lipid-induced disorders brought on by consumption of excess saturated fat. PMID- 17127674 TI - Peripheral effect of alpha-melanocyte-stimulating hormone on fatty acid oxidation in skeletal muscle. AB - To study the peripheral effects of melanocortin on fuel homeostasis in skeletal muscle, we assessed palmitate oxidation and AMP kinase activity in alpha melanocyte-stimulating hormone (alpha-MSH)-treated muscle cells. After alpha-MSH treatment, carnitine palmitoyltransferase-1 and fatty acid oxidation (FAO) increased in a dose-dependent manner. A strong melanocortin agonist, NDP-MSH, also stimulated FAO in primary culture muscle cells and C2C12 cells. However, [Glu6]alpha-MSH-ND, which has ample MC4R and MC3R agonistic activity, stimulated FAO only at high concentrations (10(-5) M). JKC-363, a selective MC4R antagonist, did not suppress alpha-MSH-induced FAO. Meanwhile, SHU9119, which has both antagonistic activity on MC3R and MC4R and agonistic activity on both MC1R and MC5R, increased the effect of alpha-MSH on FAO in both C2C12 and primary muscle cells. Small interference RNA against MC5R suppressed the alpha-MSH-induced FAO effectively. cAMP analogues mimicked the effect of alpha-MSH on FAO, and the effects of both alpha-MSH and cAMP analogue-mediated FAO were antagonized by a protein kinase A inhibitor (H89) and a cAMP antagonist ((Rp)-cAMP). Acetyl-CoA carboxylase activity was suppressed by alpha-MSH and cAMP analogues by phosphorylation through AMP-activated protein kinase activation in C2C12 cells. Taken together, these results suggest that alpha-MSH increases FAO in skeletal muscle, in which MC5R may play a major role. Furthermore, these results suggest that alpha-MSH-induced FAO involves cAMP-protein kinase A-mediated AMP-activated protein kinase activation. PMID- 17127675 TI - Reliability and validity of the Asthma Trigger Inventory applied to a pediatric population. AB - OBJECTIVE: To test the reliability and validity of the Asthma Trigger Inventory (ATI) applied to a pediatric population. METHOD: Children with asthma (N = 272, 56% male, age 7-17) and their primary caregivers answered together an asthma trigger inventory, ATI (Ritz, Steptoe, Bobb, Harris, & Edwards, 2006) developed for adults. Cronbach's alpha, principal component analysis (PCA), hierarchical regression, and correlations of the ATI subscales with skin prick tests, psychological questionnaires, and disease severity were used to assess the psychometric properties of the ATI. RESULTS: The ATI subscales demonstrated excellent reliability regardless of gender, race, socioeconomic status (SES), or age. PCA confirmed and replicated the theoretical structure of the ATI. Hierarchical multiple regressions illuminated the association of ATI subscales with demographics and asthma history. Evidence in support of construct validity was found in associations between ratings of triggering and disease severity and asthma-related quality of life (PAQLQ). Criterion validity for allergy triggering was partially supported by correlations between ATI animal allergens subscale and the cat dander skin prick test, and construct validity for emotional triggering by associations between the emotional trigger subscale score and the anxiety (STAIC) and depression (CDI, CDI-P, CDRS-R, and CBCL-I) scores. CONCLUSION: The ATI holds promise as a reliable, valid, and useful clinical and research tool to assess the type and degree of asthma triggering in a pediatric population (age 7 17) of varied gender, race, and SES. PMID- 17127676 TI - Extensions to gene set enrichment. AB - MOTIVATION: Gene Set Enrichment Analysis (GSEA) has been developed recently to capture changes in the expression of pre-defined sets of genes. We propose number of extensions to GSEA, including the use of different statistics to describe the association between genes and phenotypes of interest. We make use of dimension reduction procedures, such as principle component analysis, to identify gene sets with correlated expression. We also address issues that arise when gene sets overlap. RESULTS: Our proposals extend the range of applicability of GSEA and allow for adjustments based on other covariates. We have provided a well-defined procedure to address interpretation issues that can raise when gene sets have substantial overlap. We have shown how standard dimension reduction methods, such as PCA, can be used to help further interpret GSEA. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 17127677 TI - Mosclust: a software library for discovering significant structures in bio molecular data. AB - The R package mosclust (model order selection for clustering problems) implements algorithms based on the concept of stability for discovering significant structures in bio-molecular data. The software library provides stability indices obtained through different data perturbations methods (resampling, random projections, noise injection), as well as statistical tests to assess the significance of multi-level structures singled out from the data. AVAILABILITY: http://homes.dsi.unimi.it/~valenti/SW/mosclust/download/mosclust_1.0.tar.gz. SUPPLEMENTARY INFORMATION: http://homes.dsi.unimi.it/~valenti/SW/mosclust. PMID- 17127678 TI - GOlorize: a Cytoscape plug-in for network visualization with Gene Ontology-based layout and coloring. AB - We have implemented a graph layout algorithm that exposes Gene Ontology (GO) class structure on the network nodes. It can be used in conjunction with BiNGO plug-in to Cytoscape, which finds the GO categories over-represented in a given network. Our plug-in, named GOlorize, first highlights the class members with category-specific color-coding and then constructs an enhanced visualization of the network using a class-directed layout algorithm. AVAILABILITY: http://www.cytoscape.org/plugins2.php. SUPPLEMENTARY INFORMATION: Installation instructions and tutorial at http://www.cytoscape.org/plugins/GOlorize/GOlorizeUserGuide.pdf. PMID- 17127679 TI - QSCOP--SCOP quantified by structural relationships. AB - The database SCOP (Structural Classification Of Proteins) has become a major resource in bioinformatics and protein science. A particular strength of SCOP is the flexibility of its rules enabling the preservation of the many details spotted by experts in the classification process. Here we endow classic SCOP Families with quantified structural information and comment on the structural diversity found in the SCOP hierarchy. AVAILABILITY: Quantified SCOP (QSCOP) is available as a public WEB service. http://services.came.sbg.ac.at. PMID- 17127680 TI - Mclip: motif detection based on cliques of gapped local profile-to-profile alignments. AB - A multitude of motif-finding tools have been published, which can generally be assigned to one of three classes: expectation-maximization, Gibbs-sampling or enumeration. Irrespective of this grouping, most motif detection tools only take into account similarities across ungapped sequence regions, possibly causing short motifs located peripherally and in varying distance to a 'core' motif to be missed. We present a new method, adding to the set of expectation-maximization approaches, that permits the use of gapped alignments for motif elucidation. AVAILABILITY: The program is available for download from: http://bioinfoserver.rsbs.anu.edu.au/downloads/mclip.jar. SUPPLEMENTARY INFORMATION: http://bioinfoserver.rsbs.anu.edu.au/utils/mclip/info.php. PMID- 17127681 TI - Integrating transcription factor binding site information with gene expression datasets. AB - MOTIVATION: Microarrays are widely used to measure gene expression differences between sets of biological samples. Many of these differences will be due to differences in the activities of transcription factors. In principle, these differences can be detected by associating motifs in promoters with differences in gene expression levels between the groups. In practice, this is hard to do. RESULTS: We combine correspondence analysis, between group analysis and co inertia analysis to determine which motifs, from a database of promoter motifs, are strongly associated with differences in gene expression levels. Given a database of motifs and gene expression levels from a set of arrays, the method produces a ranked list of motifs associated with any specified split in the arrays. We give an example using the Gene Atlas compendium of gene expression levels for human tissues where we search for motifs that are associated with expression in central nervous system (CNS) or muscle tissues. Most of the motifs that we find are known from previous work to be strongly associated with expression in CNS or muscle. We give a second example using a published prostate cancer dataset where we can simply and clearly find which transcriptional pathways are associated with differences between benign and metastatic samples. AVAILABILITY: The source code is freely available upon request from the authors. PMID- 17127682 TI - A simple track structure model of ion beam radiotherapy. AB - A simple radiotherapy ion beam calculation based on the cellular track structure model, using in vitro cell survival parameters fitted from recent experimental data, is presented. The calculation represents a single-fraction ion exposure (roughly corresponding to a 2 Gy fraction of megavolt X rays) and exploits concepts used in clinical radiotherapy, such as entrance, or 'skin' ion dose. The depth distribution of cells surviving their irradiation by a beam of 385 MeV amu( 1) carbon ions is calculated over the range of the stopping ions, as a sequence of track-segments, in the continuous slowing-down approximation. An interpretation of the 'clinical relative biological effectiveness' concept is suggested. PMID- 17127684 TI - Genomewide linkage scan of hand osteoarthritis in female twin pairs showing replication of quantitative trait loci on chromosomes 2 and 19. AB - BACKGROUND AND OBJECTIVE: Until recently, there has been little agreement between conflicting results of osteoarthritis (OA) linkage. The purpose of this study was to conduct a whole-genome linkage scan to identify susceptibility loci for idiopathic hand OA in a large, population-based sample of females. METHODS: Two OA-related radiographic phenotypes DIP (distal interphalangeal joints)-OA and Tot KL (Kellgren-Lawrence score for both hands) chosen a priori were examined on 538 (269 pairs) monozygous and 1256 (628 pairs) dizygous (DZ) females. A genome-wide scan using microsatellite markers spaced 10 cM apart was performed on 1028 DZ twins. First, the heritability of the two OA phenotypes was estimated. Next, multipoint linkage analysis was conducted using a modified version of the Haseman Elston method in a generalised linear model. RESULTS: Heritability for DIP-OA and Tot-KL was found to be 47.6% and 67.4%, respectively. A genome-wide scan produced reliable evidence of significant linkage of DIP-OA on chromosome 2 at 90 cM (logarithmic odds ratio (LOD) = 2.90) and for Tot-KL on chromosome 19 at 65 cM (LOD = 4.26). These results are in agreement with data published previously. Several other significant linkage peaks were observed-for example, on chromosome 1 at 250 cM and on chromosome 3 at 30 cM-but were confirmed less reliably. CONCLUSION: This is one of the largest OA linkage studies performed to date and provides clear evidence for linkage at two quantitative trait loci (on chromosome 2 at 90 cM and on chromosome 19 at 65 cM). As the results were robust and replicated in previous smaller studies, the fine mapping of these regions is a logical next step to pinpoint potential susceptibility gene(s) of interest. PMID- 17127685 TI - Are there gender differences in severity of ankylosing spondylitis? Results from the PSOAS cohort. AB - OBJECTIVE: To examine the clinical and radiographic features in men and women in the Prospective Study of Outcomes in Ankylosing Spondylitis cohort, a large well defined cross-sectional study of patients with AS, in order to understand the influence of gender in determining the severity of ankylosing spondylitis. METHODS: Extensive clinical assessments and spine radiographs were performed in 302 men and 100 women with AS of > or = 20 years duration. Radiographs were scored using the Bath Ankylosing Spondylitis Radiographic Index Spine (BASRI spine) score (range 2-12). Functional impairment was measured by the Bath Ankylosing Spondylitis Functional Index (BASFI) and the Health Assessment Questionnaire for the Spondyloarthropathies (HAQ-S). RESULTS: Radiographic severity was worse among men. The unadjusted median BASRI-spine score for men was 10, compared with 6.5 for women (p<0.001). Functional disability, as measured by the BASFI and HAQ-S, was not different between men and women. However, after adjusting for radiographic spinal damage, women were found to report worse functioning than men at any given level of radiographic damage. Women had a slightly earlier age of disease onset; however, disease duration was identical in both groups. Women more frequently reported family histories of AS in first degree relatives and were more likely to be treated with intra-articular steroids, sulphasalazine and prednisone. CONCLUSIONS: Among patients with longstanding AS, men have more severe radiographic changes; findings of treatment differences suggest that women may have more peripheral arthritis. At any given level of radiographic damage, self-reported functional limitations were worse for women. PMID- 17127686 TI - QuantiFERON TB Gold Test: the new standard for screening of latent tuberculosis in patients with rheumatoid arthritis? PMID- 17127687 TI - Response to intramuscular methyl prednisolone in inflammatory hand pain: evidence for a targeted clinical, ultrasonographic and therapeutic approach. AB - BACKGROUND: Hand pain with stiffness is a common clinical presentation to early arthritis clinics, with outcome varying from resolution to the development of rheumatoid arthritis. OBJECTIVE: To assess the response and predictors of response to intramuscular methylprednisolone (MP) and hydroxychloroquine (HCQ) using a standardised treatment protocol. METHODS: Patients with inflammatory hand pain (IHP), defined as predominantly hand pain and morning stiffness of at least 30 min duration, received a standardised assessment prior to receiving intramuscular MP. Response (primary outcome) at 4 weeks was a 50% improvement in symptoms as perceived by the patient; responders who relapsed received repeat intramuscular MP and HCQ. RESULTS: 102 patients were recruited, of which 21% were rheumatoid factor (RF) positive, 23% had clinical synovitis, 25% had raised C reactive protein level and 55% had ultrasound-detected synovitis. 73% responded, with associated significant reductions in morning stiffness, Health Assessment Questionnaire, painful and tender joint counts, and visual analogue scores (p < or = 0.006 for all). Ultrasound-detected synovitis (p<0.001) and RF (p = 0.04), but not clinical synovitis (p = 0.74), were significantly associated with response to intramuscular MP. 86% who remained on HCQ long term reported a benefit. CONCLUSIONS: Patients with IHP have significant improvement in symptoms and function following intramuscular MP. Further placebo-controlled trials are required to assess the role of intramuscular MP and ultrasonography in managing this patient group. PMID- 17127688 TI - Investigation of respiration of individual bovine embryos produced in vivo and in vitro and correlation with viability following transfer. AB - BACKGROUND: Quantification of oxygen consumption by individual preimplantation embryos has the potential to improve embryo selection. This study investigated whether respiration rates of individual embryos are useful indicators of embryo viability. The effect of the Nanorespirometer on embryo viability was also evaluated. METHODS: The respiration rates of individual day 7 bovine in vivo- (n=44) and in vitro-produced (n=156) embryos were measured using the Nanorespirometer. In vivo-produced embryos were individually transferred to recipients. RESULTS: The respiration rates of in vivo-produced embryos increased with increasing morphological quality and stage of development (P < 0.05). Pregnancy rates on days 35 and 60 were 65 and 60%, respectively. The mean respiration rate did not differ significantly between embryos producing and not producing a pregnancy, but the transfer of embryos with respiration rates <0.78 nl/h, between 0.78 and 1.10 nl/h, and >1.10 nl/h resulted in 48, 100 and 25% pregnancy rate, respectively. The mean respiration rate of in vitro-produced embryos was higher than that of in vivo-produced embryos because of differences in the morphological quality and stage of development. CONCLUSION: The Nanorespirometer does not adversely influence embryo viability, but the sample size was too small to confirm the significance of the correlation observed between respiration rates and viability. PMID- 17127689 TI - The axis of polarity of the mouse blastocyst is specified before blastulation and independently of the zona pellucida. AB - BACKGROUND: Rather than being prepatterned, orientation of the embryonic abembryonic (Em-Ab) axis of the mouse blastocyst has been claimed to depend on the conceptus being constrained by its zona pellucida (ZP) during blastulation. This hypothesis merited closer scrutiny, because it seemed at variance with observations on living conceptuses. METHODS: Two-cell conceptuses with an oil drop injected into the lesser diameter (LD) of the ZP at the first cleavage plane were cultured until shortly before blastulation when the blastomere underlying the drop was labelled with carbocyanine dye. After removing the ZP, conceptuses were re-cultured to the blastocyst stage for recording the position along the axis of the centres of the patches of labelled cells. RESULTS: These centres showed significant bias towards the equatorial (Eq) region of the axis compared with those resulting from labelling a blastomere at random, even following softening of the ZP at the 2-cell stage. This was also true if conceptuses were denuded at the 2-cell stage and the blastomere underlying an intact second polar body (PB) labelled in morulae. CONCLUSIONS: These findings further support the view that the Em-Ab axis of the mouse blastocyst is normally prepatterned and provide no evidence of a role for the ZP in its specification. PMID- 17127690 TI - The eco- in eco-epidemiology. PMID- 17127691 TI - Lack of efficacy of Rituximab in a patient with essential mixed cryoglobulinaemia. PMID- 17127692 TI - Comment on: Parotid gland biopsy compared with labial biopsy in the diagnosis of patients with primary Sjogren's syndrome. PMID- 17127694 TI - Primary hyperaldosteronism in a patient with end-stage renal disease. PMID- 17127695 TI - Effectiveness of a chronic kidney disease clinic in achieving K/DOQI guideline targets at initiation of dialysis--a single-centre experience. AB - BACKGROUND: Limited data exist about the effects of chronic kidney disease (CKD) clinics on quality-of-care indicators in patients before initiation of dialysis. METHODS: A single-centre retrospective chart review study was conducted on all patients who initiated dialysis at the Veterans Affairs Denver Healthcare System between 2000 and 2005. Patients initiating dialysis were evaluated at the start of dialysis and 12 months after dialysis initiation, as a function of care provided by nephrologists in training (renal-hypertension clinic) vs a trained renal nurse practitioner (CKD clinic). RESULTS: Data were available for 77 patients followed in the CKD clinic and 36 in the renal-hypertension clinic. There were no major demographic differences between the cohorts at the time of clinic referral. The length of follow-up before dialysis did not differ significantly between the cohorts (10.7+/-9.8 months for the patients in the CKD clinic cohort and 13.6+/-16.0 months for the patients in the renal-hypertension clinic cohort, P=0.3299). At the initiation of dialysis, patients followed in the CKD clinic had higher haemoglobin (11.6+/-1.5 vs 10.8+/-1.7 g/dl, P=0.0239) and serum albumin (3.4+/-0.5 vs 3.0+/-0.7 g/dl, P=0.0020) concentrations. More of the CKD clinic patients had a functioning permanent vascular access (P<0.0001). The number of all-cause hospitalizations in the 12 months after initiation of dialysis was significantly lower in the CKD clinic group (P=0.0024), but no significant differences were noted in all-cause mortality. CONCLUSIONS: Our data indicate that a single experienced renal nurse practitioner, working to a protocol, is more likely to adhere to guidelines than are multiple nephrology trainees rotating through a nephrology clinic. PMID- 17127696 TI - Renal cysts and diabetes due to a heterozygous HNF-1beta gene deletion. PMID- 17127697 TI - Lack of specific binding of Shiga-like toxin (verocytotoxin) and non-specific interaction of Shiga-like toxin 2 antibody with human polymorphonuclear leucocytes. AB - BACKGROUND: After gastrointestinal infection with Shiga-like toxin (Stx) producing Escherichia coli, the toxin is transported from the intestine to the renal microvascular endothelium. This is the main target for Stx in humans. Previous studies indicated that polymorphonuclear leucocytes (PMN) could serve as carriers for Stx in the systemic circulation. As at a later stage we could not confirm these data, we performed new studies. METHODS: The binding of Stx1 to PMN was determined in vitro (isolated human PMN and whole blood) and in vivo (injection in mice). The specificity of binding of an antibody against Stx2 to PMN from patients with haemolytic uraemic syndrome (HUS) was determined. This was compared with binding to PMN from healthy controls, and patients after haemodialysis (HD) or on peritoneal dialysis (PD). Furthermore, PMN were incubated with Stx to study possible activation. RESULTS: No specific binding of Stx1 to PMN could be detected. After intravenous injection of the toxin in mice, it was not associated with PMN. The binding of an antibody against Stx2 to PMN was detected in both patients with HUS and patients after HD, but not in patients on PD. Stx was not able to activate PMN. CONCLUSIONS: PMN are not acting as transporter for Stx in the pathogenesis of HUS. The interaction of a Stx antibody with PMN from HUS patients is not specific as it can also be observed in patients after HD (possibly due to activation of the PMN). Therefore, binding of Stx antibody to PMN is not reliable as a diagnostic tool for HUS. PMID- 17127698 TI - Very early steroid withdrawal in simultaneous pancreas-kidney transplants. AB - BACKGROUND: Simultaneous pancreas-kidney (SPK) transplantation is an effective treatment for patients suffering from type 1 diabetes mellitus. Conventional immunosuppressive treatments include steroids that may induce insulin resistance and are responsible for many side effects. In de novo SPK, early withdrawal of corticosteroids may be an important issue. METHODS: A total of 24 consecutive patients with type 1 diabetes mellitus had been treated by SPK transplantation. All of them had a short induction therapy with anti-thymoglobulin (ATG) and steroids for only 4 days, association with CellCept and tacrolimus. The rate of acute rejection, graft and patient survival and side effects have been analysed. RESULTS: Patient and kidney survival was 100% and the pancreas survival was 95.6% at 1 year. The rate of acute rejection of kidney and pancreas was 4.2% and 8.3% at 6 months, respectively. The mean serum creatinine was 98.9+/-19.6 micromol/l and the mean HbA1c concentration was 5.1%+/-0.5% at 6 months. Only four patients developed a cytomegalovirus primo-infection, associated in one case with pneumonia, whereas 75% of patients developed a bacterial infection. Because of the occurrence of leucopenia and/or diarrhoea, CellCept has been dramatically decreased in 33% of cases and required the re-introduction of steroids. CONCLUSION: A short induction with ATG and steroids associated with a chronic therapy with CellCept and tacrolimus is safe and efficient in preventing acute renal rejection in SPK. PMID- 17127699 TI - The Mayo Clinic quadratic equation improves the prediction of glomerular filtration rate in diabetic subjects. AB - BACKGROUND: Although recommended, both the Cockcroft and Gault formula (CG) and the modification of diet in renal disease (MDRD) equation are not ideally predictive of glomerular filtration rate (GFR) in diabetic subjects; we tested whether the new Mayo Clinic Quadratic (MCQ) equation performed better. METHODS: In 200 diabetic subjects with a wide range of renal function, GFR was measured by 51Cr-EDTA clearance, and compared with the results of the three predictive equations by regression analysis and Bland and Altman procedures. The correlations with body mass index, age and albumin excretion rates were tested. The precisions (absolute difference as percentage), diagnostic accuracies [receiver operating characteristic (ROC) curves for the diagnosis of moderate and severe chronic kidney disease (CKD)], and the results of stratification according to the KDOQ classification were compared. RESULTS: The CG and MCQ overestimated mean GFR, whereas the MDRD underestimated it. Correlation coefficients and areas under the ROC curves were better for the MDRD and the MCQ as compared with the CG, which was biased by body weight (+30% overestimation in obese diabetic subjects). The absolute differences with true GFR were slightly lower for the MDRD than the MCQ, and both better than the CG. Both the MDRD and MCQ correctly stratified 65% of the subjects (CG: 55%, P<0.05). In contrast with the MDRD, the MCQ did not underestimate normal GFR, and its performance for stratification was uniformly good over a wide GFR range. CONCLUSIONS: In diabetic subjects, the MCQ has a similar diagnostic performance to the MDRD, but it does not underestimate normal GFR, which is an important advantage. PMID- 17127700 TI - Renal outcome after ciclosporin-induced nephrotoxicity. AB - BACKGROUND: Renal outcome after ciclosporin (CsA) is not clear in most studies involving patients with many renal comorbid conditions. We first report on renal function recovery after CsA in previously healthy kidney patients. METHODS: Uveitis patients, enroled in a unique single centre cohort follow-up study initiated in 1987, were prospectively evaluated for plasma creatinine and glomerular filtration rate (GFR) before, during (>2 years) and after (>6 months) CsA therapy. We hypothesized that CsA alters renal function progressively over time according to two additive exponential components (irreversible and reversible) and used a mixed linear model with exponential speed parameters maximizing the likelihood. RESULTS: Twenty-seven patients treated for 60+/-34 months (CsA 5.1+/-2.5 mg/kg/day) were followed up for 56+/-42 months after CsA withdrawal. Baseline creatinine was 0.92+/-0.15 mg/dl. The reversible effect of CsA was quantified as a 0.11+/-0.07 mg/dl increase in creatinine/100 mg CsA/day (P<0.001) and a 6.0+/-3.7 ml/min/1.73 m(2) decrease in GFR/100 mg CsA/day (P<0.0001). The irreversible effect was quantified as a 0.03+/-0.05 mg/dl increase in creatinine/100 g cumulative CsA received (P<0.007) and a decrease of 3.3+/-3.9 ml/min/1.73 m(2) GFR/100 g CsA. CONCLUSIONS: Although significant decrease in GFR is induced by low-dose CsA therapy in previously healthy kidney patients, renal function recovery is possible after CsA withdrawal and best predicted by CsA daily dosage. Irreversible loss in GFR is correlated to cumulated CsA exposure. The lowest CsA dosage and shortest exposure time effect as well as unlimited renal monitoring are required in order to provide the best long-term renal outcome. PMID- 17127701 TI - Efficacy of percutaneous ethanol injection therapy (PEIT) is related to the number of parathyroid glands in haemodialysis patients with secondary hyperparathyroidism. AB - BACKGROUND: Percutaneous ethanol injection therapy (PEIT) is used for advanced secondary hyperparathyroidism. We investigated the efficacy, remission period and risk of relapse to determine the effect of the number of hyperplastic glands and other factors on the therapeutic effect of PEIT. METHODS: We studied 321 patients divided into two groups: effective [serum corrected calcium (cCa) level < or =10.5 mg/dl and serum intact parathyroid hormone (iPTH) level < or =250 pg/ml], and ineffective (failed to achieve the target levels). Advanced hyperplasia was defined as an estimated volume > or =0.5 cm(3) on ultrasonography. RESULTS: PEIT was effective in 201 patients (62.6%), in whom serum iPTH levels dropped from 603+/-292 to 183+/-62 pg/ml (ng/l) and serum cCa levels from 10.7+/-0.8 to 10.1+/ 0.5 mg/dl. Univariate analysis identified age, the number of hyperplastic glands and iPTH level as factors related to the efficacy of PEIT. The odds ratio for success vs failure by multivariate analysis was 0.55 times for the number of hyperplastic glands > or =0.5 cm(3) (> or =2 vs 0,1) and 0.29 times for iPTH (> or =500 vs <500 pg/ml). Using the Kaplan-Meier method, the number of hyperplastic glands > or =0.5 cm(3) (> or =2 vs 0,1) was a factor affecting the remission period, with a remission significantly longer seen in the group with one hyperplastic gland (P=0.0025). CONCLUSIONS: Superior results in efficacy rate, remission period and risk of relapse are obtained when PEIT is restricted to patients with one hyperplastic gland > or =0.5 cm(3). PMID- 17127703 TI - p53 independent induction of PUMA mediates intestinal apoptosis in response to ischaemia-reperfusion. AB - BACKGROUND: The small intestine is highly sensitive to ischaemia-reperfusion (I/R) induced injury which is associated with high morbidity and mortality. Apoptosis, or programmed cell death, is a major mode of cell death occurring during I/R induced injury. However, the mechanisms by which I/R cause apoptosis in the small intestine are poorly understood. p53 upregulated modulator of apoptosis (PUMA) is a p53 downstream target and a member of the BH3-only group of Bcl-2 family proteins. It has been shown that PUMA plays an essential role in apoptosis induced by a variety of stimuli in different tissues through a mitochondrial pathway. AIMS: The role of PUMA in I/R induced injury and apoptosis in the small intestine was investigated. The mechanisms by which PUMA is regulated in I/R induced intestinal apoptosis were also studied. METHODS: Ischaemia was induced by superior mesenteric artery occlusion in the mouse small intestine. Induction of PUMA in response to ischaemia alone, or ischaemia followed by reperfusion (I/R), was examined. I/R induced intestinal apoptosis and injury were compared between PUMA knockout and wild-type mice. The mechanisms of I/R induced and PUMA mediated apoptosis were investigated through analysis of caspase activation, cytosolic release of mitochondrial cytochrome c and alterations of the proapoptotic Bcl-2 family proteins Bax and Bak. To determine whether PUMA is induced by reactive oxygen species and/or reactive nitrogen species generated by I/R, superoxide dismutase (SOD) and N-nitro-l-arginine methyl ester (L-NAME) were used to treat animals before I/R. To determine whether p53 is involved in regulating PUMA during I/R induced apoptosis, PUMA induction and apoptosis in response to I/R were examined in p53 knockout mice. RESULTS: PUMA was markedly induced following I/R in the mucosa of the mouse small intestine. I/R induced intestinal apoptosis was significantly attenuated in PUMA knockout mice compared with that in wild-type mice. I/R induced caspase 3 activation, cytochrome c release, Bax mitochondrial translocation and Bak multimerisation were also inhibited in PUMA knockout mice. SOD or L-NAME significantly blunted I/R induced PUMA expression and apoptosis. Furthermore, I/R induced PUMA expression and apoptosis in the small intestine were not affected in the p53 knockout mice. CONCLUSIONS: Our data demonstrated that PUMA is activated by oxidative stress in response to I/R to promote p53 independent apoptosis in the small intestine through the mitochondrial pathway. Inhibition of PUMA is potentially useful for protecting against I/R induced intestinal injury and apoptosis. PMID- 17127702 TI - Adenovirus-mediated expression of BMP-7 suppresses the development of liver fibrosis in rats. AB - BACKGROUND: Liver cirrhosis, which is caused by the accumulation of extracellular matrix materials, is a serious clinical problem that can progress to hepatic failure. Transforming growth factor-beta (TGFbeta) plays a pivotal role in extracellular matrix production, but bone morphogenetic protein (BMP)-7, a member of the TGFbeta superfamily, can antagonise the fibrogenic activity of TGFbeta. AIM: In this study, we examined whether adenovirus-mediated overexpression of BMP 7 (Ad-BMP-7) antagonised the effect of TGFbeta in vitro and in vivo. METHODS AND RESULTS: In primary cultured rat stellate cells and the LX-2 human stellate cell line, induction of BMP-7 by Ad-BMP-7 infection decreased the expression of collagen 1A2 mRNA and smooth muscle alpha-actin in the presence or absence of TGFbeta, via Smad 1/5/8 phosphorylation. BMP-7 triggered the mRNA expression of inhibitors of differentiation 2 (Id2) in LX-2. Although endogenous expression of BMP-7 was hardly detectable, Smad1 and Id2 overexpression increased BMP-7 expression in LX-2. A liver fibrosis model was induced by the repetitive intraperitoneal injection of thioacetamide (200 mg/kg body weight) twice per week for up to 7 weeks. In rats administered Ad-BMP-7 via the tail vein, hydroxyproline content and the areas stained by Sirius red dye in the liver were significantly reduced compared to controls. Ad-Id2 also reduced fibrosis. CONCLUSION: These data demonstrate that BMP-7, Smad 1/5/8 and Ids interact to antagonise hepatic fibrogenesis. PMID- 17127704 TI - Predicting response to peginterferon alpha-2a, lamivudine and the two combined for HBeAg-negative chronic hepatitis B. AB - OBJECTIVE: In a trial of patients with hepatitis B e antigen (HBeAg)-negative chronic hepatitis B, 24 week post-treatment biochemical and virological response rates with peginterferon alpha-2a with or without lamivudine were significantly higher than with lamivudine alone. The effect of pre-treatment factors on post treatment responses was investigated. METHODS: Multivariate analyses were performed using available data from 518 patients treated with peginterferon alpha 2a with or without lamivudine, or with lamivudine alone. A post-treatment response was defined as alanine aminotransferase (ALT) normalisation and hepatitis B virus (HBV) DNA level of <20,000 copies/ml. RESULTS: In logistic regression analyses across all treatment arms, peginterferon alpha-2a (with or without lamivudine) therapy, younger age, female gender, high baseline ALT, low baseline HBV DNA and HBV genotype were identified as significant predictors of combined response at 24 weeks post-treatment. In the peginterferon alpha-2a and lamivudine monotherapy arms, patients with genotypes B or C had a higher chance of response than genotype D infected patients (p<0.001), the latter responding better to the combination than to peginterferon alpha-2a monotherapy (p = 0.015). At 1 year post-treatment, response rates by intention-to-treat analysis were 19.2% for the peginterferon alpha-2a, 19.0% for the combination, and 10.0% for the lamivudine groups, with genotypes B or C associated with a sustained combined response to peginterferon alpha-2a with or without lamivudine therapy. CONCLUSIONS: Baseline ALT and HBV DNA levels, patient age, gender, and infecting HBV genotype significantly influenced combined response at 24 weeks post treatment, in patients treated with peginterferon alpha-2a and/or lamivudine. At 1 year post-treatment HBV genotype was significantly predictive of efficacy for patients treated with peginterferon alpha-2a with or without lamivudine. PMID- 17127705 TI - Deficiency of invariant natural killer T cells in coeliac disease. AB - BACKGROUND: Immunoregulatory invariant natural killer (iNK) T cells rapidly produce interleukin (IL)-4 and other cytokines that suppress a Th1 response and are deficient in some autoimmune diseases. AIM: The aim of this study was to investigate any deficiency of iNK T cells in coeliac disease. METHODS: Blood was collected from 86 subjects with coeliac disease and from 152 healthy control subjects for investigation of Valpha24+ T cells by flow cytometry. iNK T cells were assessed by Valpha24 and alpha-galactosylceramide/CD1d tetramer markers in 23 normal controls and 13 subjects with coeliac disease. Intracellular IL-4 was measured after anti-CD3 antibody stimulation. Duodenal biopsies were obtained in a subgroup of subjects with coeliac disease and control subjects for Valpha24 mRNA expression using relative PCR and for Valpha24+ T cells by immunofluorescence. RESULTS: The mean numbers of circulating Valpha24+ T cells and iNK T cells in coeliac disease were 27% (p<0.001) and 16% (p<0.001), respectively, of levels in control subjects. After in vitro anti-CD3 stimulation, numbers of IL-4+ producing iNK T cells from subjects with coeliac disease were unchanged but increased by 21% in control subjects. In subjects with coeliac disease, Valpha24 mRNA intestinal expression was reduced to 17% (p<0.001) by relative PCR and numbers of intestinal Valpha24+ T cells were 16% (p<0.01) of levels in control subjects. CONCLUSIONS: We conclude that Valpha24+ T cells and iNK T cells are deficient in coeliac disease. We speculate that this deficiency could contribute to the failure of immunological oral tolerance that seems to underlie this disease. PMID- 17127706 TI - Obesity increases oesophageal acid exposure. AB - BACKGROUND: Obesity has been associated with gastro-oesophageal reflux disease (GERD); however, the mechanism by which obesity may cause GERD is unclear. AIM: To examine the association between oesophageal acid exposure and total body or abdominal anthropometric measures. METHODS: A cross-sectional study of consecutive patients undergoing 24 h pH-metry was conducted. Standardised measurements of body weight and height as well as waist and hip circumference were obtained. The association between several parameters of oesophageal acid exposures and anthropometric measures were examined in univariate and multivariate analyses. RESULTS: 206 patients (63% women) with a mean age of 51.4 years who were not on acid-suppressing drugs were enrolled. A body mass index (BMI) of >30 kg/m(2) (compared with BMI<25 kg/m(2)) was associated with a significant increase in acid reflux episodes, long reflux episodes (>5 min), time with pH<4, and a calculated summary score. These significant associations have affected total, postprandial, upright and supine pH measurements. Waist circumference was also associated with oesophageal acid exposure, but was not as significant or consistent as BMI. When adjusted for waist circumference by including it in the same model, the association between BMI>30 kg/m(2) and measures of oesophageal acid exposure became attenuated for all, and not significant for some, thus indicating that waist circumference may mediate a large part of the effect of obesity on oesophageal acid exposure. CONCLUSIONS: Obesity increases the risk of GERD, at least partly, by increasing oesophageal acid exposure. Waist circumference partly explains the association between obesity and oesophageal acid exposure. PMID- 17127707 TI - Branch-duct intraductal papillary mucinous neoplasms of the pancreas: to operate or not to operate? AB - BACKGROUND: Branch-duct intraductal papillary mucinous neoplasms (BD-IPMNs) of the pancreas are reported to be less aggressive than the main-duct type. Hence, less aggressive treatment has been proposed for the former. AIM: To evaluate the effectiveness of a follow-up protocol for BD-IPMNs. DESIGN: Prospective study. SETTING: An academic tertiary referral centre. PATIENTS: From 2000 to 2003, 109 patients with BD-IPMNs underwent trans-abdominal ultrasound and magnetic resonance cholangiopancreatography with secretin. Patients who presented malignancy-related parameters (size >3.5 cm, nodules, thick walls, carbohydrate antigen 19.9 level >25 U/l, recent-onset or worsened diabetes) and/or complained of symptoms were submitted to surgery (arm A). All asymptomatic patients without suspicion of malignancy were followed up according to a 6-month clinical radiological protocol (arm B). MAIN OUTCOME MEASURES: The effectiveness of conservative management of BD-IPMNs. RESULTS: 20 (18.3%) patients underwent surgery (arm A); pathological diagnosis of BD-IPMNs was always confirmed. 89 (81.7%) patients were followed up for a median of 32 months (arm B); of these, 57 (64%) patients had multifocal disease. After a mean follow-up of 18.2 months, 5 (5.6%) patients showed an increase in lesion size and underwent surgery. The pathological diagnosis was branch-duct adenoma in three patients and borderline adenoma in two. CONCLUSIONS: Surgery is indicated in <20% of cases of BD-IPMNs, and, in the absence of malignancy-related parameters, careful non-operative management seems to be safe and effective in asymptomatic patients. Although observation for a longer time is needed to confirm these results, our findings support the guidelines recently recommended by the International Association of Pancreatology. PMID- 17127708 TI - Characteristics, outcomes, and predictors of 1-year mortality in patients hospitalized for acute heart failure. AB - AIMS: Acute heart failure (AHF) is associated with poor prognosis and requires recurrent hospitalizations. However, studies on AHF characteristics, treatment, and prognostic factors are few. Our aim was to investigate the characteristics, treatment, and 1-year prognosis of AHF and identify prognostic factors in different clinical groups. METHODS AND RESULTS: We conducted a prospective multicentre study with 620 patients hospitalized due to AHF; mean age 75.1 (10.4) years, 50% male. Half of the patients had new-onset heart failure. Acute congestion (63.5%) and pulmonary oedema (26.3%) were the most common clinical presentations. Left ventricular ejection fraction (LVEF) was reported in two thirds of patients. Half of these had preserved systolic function (LVEF> or =45%). At discharge, 86% of patients had beta-blockers and 76% either ACE inhibitors or angiotensin receptor blockers in use. The 12-month all-cause mortality was 27.4%. We identified several clinical and biochemical prognostic risk factors in univariate analysis. Independent predictors of 1-year mortality were older age, male gender, lower systolic blood pressure (SBP) on admission, C reactive protein, and serum creatinine >120 micromol/L. CONCLUSION: We present the characteristics and prognosis of an unselected population of AHF patients. One-year mortality is high, and independent clinical risk factors include age, male gender, lower SBP on admission, C-reactive protein, and renal dysfunction. PMID- 17127709 TI - Heart-type fatty acid-binding protein permits early risk stratification of pulmonary embolism. AB - AIMS: We investigated the value of a novel early biomarker, heart-type fatty acid binding protein (H-FABP), in risk stratification of patients with acute pulmonary embolism (PE). METHODS AND RESULTS: We prospectively included 107 consecutive patients with confirmed PE. The endpoints were (i) PE-related death or major complications and (ii) overall 30-day mortality. Overall, 29 patients (27%) had abnormal (>6 ng/mL) H-FABP levels at presentation. Of those, 12 (41%) had a complicated course, whereas all patients with normal baseline H-FABP had a favourable 30-day outcome (OR, 71.45; P<0.0001). At multivariable analysis, H FABP (P<0.0001), but not cardiac troponin T (P=0.13) or N-terminal pro-brain natriuretic peptide (P=0.36), predicted an adverse outcome. Evaluation of a strategy combining biomarker testing with echocardiography revealed that patients with a negative H-FABP test had an excellent prognosis regardless of echocardiographic findings. In contrast, patients with a positive H-FABP test had a complication rate of 23.1% even in the presence of a normal echocardiogram, and this rose to 57.1% if echocardiography also demonstrated right ventricular dysfunction (OR vs. a negative H-FABP test, 5.6 and 81.4, respectively). CONCLUSION: H-FABP is a promising early indicator of right ventricular injury and dysfunction in acute PE. It may help optimize risk stratification algorithms and treatment strategies. PMID- 17127710 TI - Lack of efficacy and cost-effectiveness of drug-eluting stents. PMID- 17127711 TI - The pituitary tumor-transforming gene promotes angiogenesis in a mouse model of follicular thyroid cancer. AB - Overexpression of the pituitary tumor-transforming gene (PTTG) has been associated with tumorigenesis. In a mouse model that spontaneously develops follicular thyroid cancer (FTC) with distant metastasis (TRbetaPV mouse), PTTG is overexpressed, similar to human thyroid cancer. To evaluate the role of PTTG in thyroid carcinogenesis, we studied the offspring of TRbetaPV mice with mice lacking PTTG (PTTG(-/-) mice). The thyroids of TRbeta(PV/PV) PTTG(-/-) mice were significantly smaller than TRbeta(PV/PV) mice. Ki-67 staining showed a decrease in thyroid proliferation in TRbeta(PV/PV) PTTG(-/-) mice. Our evaluation of the Rb-E2F pathway, a central mediator of cell growth, found that TRbeta(PV/PV) PTTG( /-) mice exhibited a decrease in protein levels of phosphorylated Rb along with an elevation of the cdk inhibitor p21. Histological examination documented no difference in FTC occurrence between TRbeta(PV/PV) and TRbeta(PV/PV) PTTG(-/-) mice, which indicates that PTTG removal does not prevent the initiation of FTC. However, TRbeta(PV/PV) PTTG(-/-) mice had a significant decrease in vascular invasion and less development of lung metastasis as they progressively aged. CD31 staining also showed a decrease in vessel density in TRbeta(PV/PV) PTTG(-/-) versus TRbeta(PV/PV) thyroids. Given the decreased vascular invasion in the PTTG knockout mice, we studied genes involved in angiogenesis. Real-time reverse transcription-polymerase chain reaction showed a consistent decrease in pro angiogenic factors, fibroblast growth factor (FGF2), its receptor FGFR1 and vascular endothelial growth factor. Our results highlight the dual roles of PTTG as a regulator of thyroid growth and contributor to tumor progression. The separation of the pathways regulating cell proliferation, tumor initiation and tumor progression should direct future therapeutic options. PMID- 17127712 TI - Urinary 8-oxodeoxyguanosine, aflatoxin B1 exposure and hepatitis B virus infection and hepatocellular carcinoma in Taiwan. AB - To evaluate the role of oxidative stress and aflatoxin exposure on risk of hepatocellular carcinoma (HCC), a case-control study nested within a community based cohort was conducted in Taiwan. Baseline urine samples, collected from a total of 74 HCC cases and 290 matched controls, were used to determine by enzyme linked immunosorbent assays the level of urinary excretion of 8-oxodeoxyguanosine (8-oxodG), a biomarker of oxidative DNA damage and urinary aflatoxin B(1) metabolites, a biomarker of aflatoxin exposure. Multivariate-adjusted linear regression analysis showed that urinary aflatoxin metabolites and gender were significantly associated with level of urinary 8-oxodG among controls. Moreover, after adjustments for potential confounding factors, there was a statistically significant positive dose-response relationship between levels of urinary 8-oxodG and urinary aflatoxin metabolites (P < 0.0001). However, when compared with subjects in the lowest quartile of 8-oxodG, there was a decrease in risk of HCC, with adjusted odds ratios (ORs) of 0.8 [95% confidence interval (CI) 0.3-2.0], 0.7 (95% CI 0.3-2.0) and 0.7 (95% CI 0.2-1.7) for subjects in the second, third and fourth quartile, respectively. The combination of level of urinary 8-oxodG below the median and hepatitis B virus infection resulted in an OR of 11.4 (95% CI 3.9-33.3), compared with those with urinary 8-oxodG above the median and hepatitis B virus surface antigen negative. These results suggest that elevated levels of urinary 8-oxodG may be related to increasing level of aflatoxin exposure but may also indicate enhanced repair of oxidative DNA damage and therefore lower risk of HCC. PMID- 17127713 TI - Overexpression of iron regulatory protein 1 suppresses growth of tumor xenografts. AB - Iron is essential for proliferation of normal and neoplastic cells. Cellular iron uptake, utilization and storage are regulated by transcriptional and post transcriptional mechanisms. We hypothesized that the disruption of iron homeostasis may modulate the growth properties of cancer cells. To address this, we employed H1299 lung cancer cells engineered for tetracycline-inducible overexpression of the post-transcriptional regulator iron regulatory protein 1 (IRP1). The induction of IRP1 (wild-type or the constitutive IRP1(C437S) mutant) did not affect the proliferation of the cells in culture, and only modestly reduced their efficiency to form colonies in soft agar. However, IRP1 dramatically impaired the capacity of the cells to form solid tumor xenografts in nude mice. Tumors derived from IRP1-transfectants were <20% in size compared to those from parent cells. IRP1 coordinately controls the expression of transferrin receptor 1 (TfR1) and ferritin by binding to iron-responsive elements (IREs) within their mRNAs. Biochemical analysis revealed high expression of epitope tagged IRP1 in tumor tissue, which was associated with a profound increase in IRE binding activity. As expected, this response misregulated iron metabolism by increasing TfR1 levels. Surprisingly, IRP1 failed to suppress ferritin expression and did not affect the levels of the iron transporter ferroportin. Our results show that the overexpression of IRP1 is associated with an apparent tumor suppressor phenotype and provide a direct regulatory link between the IRE/IRP system and cancer. PMID- 17127714 TI - Epidermal stem and progenitor cells in murine epidermis accumulate UV damage despite NER proficiency. AB - Ultraviolet (UV) radiation induces cyclobutane pyrimidine dimers (CPDs) and (6-4) photoproducts (6-4PPs) in DNA, which through gene mutations (e.g. in P53) may lead to skin carcinogenesis. Upon chronic low-level UV exposure, certain basal cells in mouse epidermis were reported to accumulate CPDs. These observations raised questions on whether these cells were fully DNA-repair deficient, and whether they were stem or progenitor cells, as suggested by their long residence time. We found that CPD-retaining basal cells (CRBCs) in SKH-1 hairless mice were repair proficient as accumulation of (6-4)PP, which is a hallmark for complete nucleotide excision repair-deficiency in rodents, was not observed. Accumulation of 6-4PP as well as CPD did, however, occur in basal cells in the epidermis of DNA repair-deficient Xpc-/- mice. Chronic UV exposure of DDB2 transgenic mice and DDB2 knockout mice revealed that the occurrence of CRBCs was inversely correlated with DDB2-expression, indicating that a boost in DNA repair lowered CPD accumulation. Stem cells are quiescent cells and can be identified as 5-bromo-2' deoxyuridine-label retaining cells (BrdU-LRCs). Induction of BrdU-LRCs followed by chronic UV irradiation showed that all BrdU-label retaining stem cells were also CPD-retaining cells. As most CRBCs were not BrdU-labeled we surmized that these cells must include BrdU-negative stem cells and early progenitor cells. In confirmation of the latter, we found that CRBCs occurred among MTS24+ hair follicle progenitor cells. These findings provide the first evidence that epidermal stem and progenitor cells are prone to the accumulation of UV-induced DNA-damage and can be a prominent target in skin carcinogenesis. PMID- 17127715 TI - Interplay between Helicobacter pylori and host gene polymorphisms in inducing oxidative DNA damage in the gastric mucosa. AB - Infection by Helicobacter pylori is the most important risk factor for gastric cancer. However, only a small fraction of colonized individuals, representing at least half of the world's population, develop this malignancy. In order to shed light on host-microbial interactions, gastric mucosa biopsies were collected from 119 patients suffering from dyspeptic symptoms. 8-Hydroxy-2'-deoxyguanosine (8 oxo-dG) levels in the gastric mucosa were increased in carriers of H.pylori, detected either by cultural method or by polymerase chain reaction, and were further increased in subjects infected with strains positive for the cagA gene, encoding the cytotoxin-associated protein, cagA. Oxidative DNA damage was more pronounced in males, in older subjects, and in H.pylori-positive subjects suffering from gastric dysplasia. Moreover, 8-oxo-dG levels were significantly higher in a small subset of subjects having a homozygous variant allele of the 8 oxoguanosine-glycosylase 1 (OGG1) gene, encoding the enzyme removing 8-oxo-dG from DNA. Conversely, they were not significantly elevated in glutathione S transferase M1 (GSTM1)-null subjects. Thus, both bacterial and host gene polymorphisms affect oxidative stress and DNA damage, which is believed to represent a key mechanism in the pathogenesis of gastric cancer. The interplay between bacterial and host gene polymorphisms may explain why gastric cancer only occurs in a small fraction of H.pylori-infected individuals. PMID- 17127716 TI - Biological assays and genomic analysis reveal lipoic acid modulation of endothelial cell behavior and gene expression. AB - Lipoic acid (LA) is a sulfated antioxidant produced physiologically as a coenzyme of the pyruvate dehydrogenase complex; it is currently used for treatment of non insulin-dependent diabetes to favor the cellular uptake of glucose. We have previously described the angiopreventive potential of molecules sharing common features with LA: N-acetyl cysteine, epigallocatechin-3-gallate and xanthohumol. To expand these studies, we have tested the capacity of LA to modulate angiogenesis in tumor growth using a Kaposi's sarcoma model. Endothelial cells exposed to LA displayed a dose-dependent reduction of cell migration and a time dependent modulation of the phosphorylation of key signaling molecules. In vivo, LA efficiently repressed angiogenesis in matrigel plugs and KS-Imm tumor growth. We analyzed modulation of gene expression in endothelial cells treated with LA for 5 h (early response), finding a mild anti-apoptotic, antioxidant and anti inflammatory response. A group of LA-targeted genes was selected to perform real time polymerase chain reaction time-lapse experiments. The long-term gene regulation (48 h and 4 days) shows higher rates of modulation as compared with the array data, confirming that LA is able to switch the regulation of several genes linked to cell survival, inflammation and oxidative stress. LA induced the production of tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL) in KS-Imm and activin-A in KS-Imm and endothelial cells; these factors show anti-angiogenic activity in vivo contributing to explain the inhibitory effect of LA on neovascularization. According to our data, LA has promising anti angiogenic properties, though its influence on central metabolic pathways should suggest more caution about its widespread and not prescribed use at pharmacological doses. PMID- 17127718 TI - Dietary effects of soy isoflavones daidzein and genistein on 7,12 dimethylbenz[a]anthracene-induced mammary mutagenesis and carcinogenesis in ovariectomized Big Blue transgenic rats. AB - The major constituents of isoflavones, daidzein (DZ) and genistein (GE) are known to interact with the alpha and beta estrogen receptors (ERalpha/beta) in several tissues including mammary. In this study, we used ovariectomy (OVX) to model menopause and determined the effects of DZ, GE or 17beta-estradiol (E2) exposures on chemically induced mutagenesis and carcinogenesis in the mammary glands of female Big Blue (BB) transgenic rats. The rats were fed control diet containing the isoflavones and E2 and treated with a single oral dose of 7,12 dimethylbenz[a]anthracene (DMBA) at PND 50. Animals were sacrificed at 16 or 20 weeks post-carcinogen treatment to assess mutant frequencies (MFs) and histopathological parameters, respectively. The isoflavones or E2 supplementation alone resulted in modest increases in the lacI MF that were not significantly different from the MFs measured in rats fed the control diet alone. DMBA exposure, however, induced significant increases in the lacI MFs in the mammary of both OVX and ovary intact (INT) rats and Hprt MFs in spleen lymphocytes (P1400 micro g/L. Tissue selenium levels show a complex pattern and significant elevations in organs such as kidney are not always indicative of toxicity. As with many trace elements, measuring selenium concentrations in body fluids and tissues tends to be easier than understanding what the results mean. PMID- 17127728 TI - Morphoproteomic demonstration of constitutive nuclear factor-kappaB activation in glioblastoma multiforme with genomic correlates and therapeutic implications. AB - Glioblastoma multiforme (GBM) presents a major challenge to neurosurgeons, neuro oncologists, and radiation therapists by virtue of its location with a blood brain barrier, chemoradioresistance, highly malignant phenotype, and angiogenic potential. Because nuclear factor-kappaB (NF-kappaB) can transcriptionally activate genes leading to the synthesis of anti-apoptotic, chemoresistant, growth promoting, and angiogenic proteins; we assessed the state of activation of NF kappaB in 6 GBM cases at diagnosis. Morphoproteomic analysis confirmed the constitutive activation of NF-kappaB by demonstrating the phosphorylation (p) and nuclear translocation of p-NF-kappaBp65 (Ser 536) in these cases. This observation coincides with (a) previous immunohistochemical findings showing nuclear translocation of total p65, (b) demonstration of NF-kappaB DNA binding activity, (c) the results of electrophoretic mobility shift assays, and (d) existing genomic data in GBM. Furthermore, such constitutive activation of the NF kappaB pathway helps to explain some of the tumor biology and supports the incorporation of NF-kappaB pathway inhibitors into the treatment of GBM. PMID- 17127729 TI - Age-associated changes in hearts of male Fischer 344/Brown Norway F1 rats. AB - Aging is associated with left ventricular hypertrophy, dilatation, and fibrosis of the heart. The Fischer 344/Brown Norway F1 (F344/BNF1) rat is recommended for age-related studies by the National Institutes on Aging because this hybrid rat lives longer and has a lower rate of pathological conditions than inbred rats. However, little is known about age-associated changes in cardiac and aortic function and structure in this model. This study evaluated age-related cardiac changes in male F344/BNF1 rats using ECHO, gross, and microscopic examinations. Rats aged 6-, 30-, and 36-mo were anesthetized and two-dimensional ECHO measurements, two-dimensional guided M-mode, Doppler M-mode, and other recordings from parasternal long- and short-axis views were obtained using a Phillips 5500 ECHO system with a 12 megahertz transducer. Hearts and aortas from sacrificed rats were evaluated grossly and microscopically. The ECHO studies revealed persistent cardiac arrhythmias (chiefly PVCs) in 72% (13/18) of 36-mo rats, 10% (1/10) of 30-mo rats, and none in 6-mo rats (0/16). Gross and microscopic studies showed left ventricular (LV) dilatation, borderline to mild hypertrophy, and areas of fibrosis that were common in 36-mo rats, less evident in 30-mo rats, and absent in 6-mo rats. Aging was associated with mild to moderate decreases of LV diastolic and systolic function. Thus, male F344/BN F1 rats demonstrated progressive age-related (a) decline in cardiac function (diastolic and systolic indices), (b) LV structural changes (chamber dimensions, volumes, and wall thicknesses), and (c) persistent arrhythmias. These changes are consistent with those in humans. The noninvasive ECHO technique offers a means to monitor serial age-related cardiac failure and therapeutic responses in the same rats over designated time intervals. PMID- 17127730 TI - Ratio of bone marrow reticulocytes to peripheral corrected reticulocytes for evaluating ineffective erythropoiesis. PMID- 17127732 TI - Imported platelets demonstrate decreased pH and glucose by reagent strip testing when compared to locally derived platelets. AB - The most common infectious risk from blood transfusion in the United States is bacterial contamination of platelet components. Although detection of bacterially contaminated platelet components is best achieved with a culture system, AABB standards permit alternatives including the use of staining methods or reagent strips. In this study, 13,216 consecutive platelet components were screened using reagent strips for evidence of the presence of bacteria. Testing was performed immediately prior to release of the platelet components for transfusion; 10,836 were collected locally and 2,380 were imported from other blood banks. A mix of whole-blood-derived and apheresis products was included. If either the glucose concentration was <250 mg/ml or the pH was <7.0, the platelet component was quarantined and a specimen was obtained for Gram's stain and culture. Every transfused platelet component that was associated with a reported transfusion reaction was also tested by Gram's stain and culture. Overall, 1.47% of imported platelet components were reactive while only 0.12% of locally collected platelets were reactive. Of 48 reactive platelet components, 44 were tested by Gram's stain and culture. None was found to be bacterially contaminated. In summary, imported platelet components were significantly more likely to be falsely reactive by reagent strip screening as compared to locally prepared platelet components. PMID- 17127733 TI - Reducing substances in urine: a paradigm for changes in a standard test. AB - Detection of reducing substances in urine has been a standard laboratory procedure for about 50 yr. It is used as a screening test for inborn errors of carbohydrate metabolism. Although the test has poor specificity and most states perform mandatory newborn screening for the common genetic defects, most clinical laboratories still perform this as a reflex test on all pediatric urine samples. We suggest that laboratories should perform this test only at the specific order of a physician and that they should review their test menu frequently to delete tests that no longer have a clinical rationale. PMID- 17127734 TI - Asymmetric dimethylarginine and nitric oxide levels as signs of endothelial dysfunction in Behcet's disease. AB - Behcet's disease (BD) has been known for many years, yet the etiology of the systemic vasculitis remains unknown. Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide (NO) synthase. ADMA is involved in endothelial dysfunction in various vascular diseases and its level in BD is unclear. This study was performed to evaluate the relationship between ADMA and NO levels in plasma of patients with BD. There were 3 groups of 30 subjects: (a) controls, (b) BD patients with mucocutaneous involvement, and (c) BD patients with vascular involvement. Plasma NO levels were assayed by spectrophotometry and plasma ADMA levels were assayed by an ELISA test. Plasma ADMA levels were higher in both groups of BD patients than in the controls; the ADMA levels were higher in the BD patients with vascular involvement than in the mucocutaneous group. Plasma NO levels were lower in both groups of BD patients than in controls; plasma NO levels were lower in the BD patients with vascular involvement than in mucocutaneous group. In the combined groups of 60 BD patients, there was significant inverse correlation between the plasma concentrations of ADMA and NO (r = -0.570, p <0.001). Plasma lipid profiles did not differ significantly between the BD patients and the controls. These results are evidence for increased plasma ADMA levels and decreased plasma NO levels as risk factors for cardiovascular events in BD patients. Inhibition of NO synthesis by ADMA may contribute to vascular involvement in BD. PMID- 17127735 TI - Antitumor and normal cell protective effect of PKC412 in the athymic mouse model of ovarian cancer. AB - N-benzoyl-staurosporine (PKC412) is a selective inhibitor of protein kinase C, and it inhibits the growth of human cancer cells. In this study, we examined the antitumor effect of PKC412, given singly and in combination with paclitaxel, on tumor regression and chemotherapeutic side effects by assessing tumor burden and cytokine production responses in vivo. Twenty-six nude mice intraperitoneally inoculated with SKOV3 cells were treated differently in 4 treatment groups: PKC412 plus paclitaxel (n = 7), paclitaxel-only (n = 6), PKC412-only (n = 6), and controls (n = 7). At autopsy, we found that PKC412 itself slightly reduced the mass of tumor but did not fully inhibit tumor formation. The incidence of evident disease was decreased when PKC412 was combined with paclitaxel (43%). From the body weight of the tumor-bearing mice, we observed that PKC412 plus paclitaxel treated mice were less wasted than paclitaxel-only treated mice (18.1 g vs 22.4 g, p = 0.001). We measured intracellular TNFalpha, IFNgamma, IL-4, and IL-10 in stimulated mouse splenocytes using flow cytometry to determine if PKC412 inhibited cytokine production in T cells. TNFalpha, IFNgamma, and IL-10 production were all significantly inhibited in the paclitaxel-treated mice. The inhibitory effects on cytokine production by paclitaxel were compensated with PKC412 combination (p = 0.008, 0.035, 0.014, respectively). From this study, we deduce that PKC412 may have clinical applications in promoting tumor regression in ovarian cancer when combined with paclitaxel. Moreover, PKC412 is able to prevent weight loss and immunosuppression induced by paclitaxel because it rescues normal proliferating cells from cytotoxic effects. PMID- 17127736 TI - An immunoglobulin A1 that inhibits lactate dehydrogenase activity, with reversal of inhibition by addition of NADH. AB - We discovered a patient with low serum lactate dehydrogenase (LD) activity and an abnormal LD isozyme pattern. We analyzed the patient's LD inhibitor using electrophoresis, affinity chromatography, and immunochemical technologies. The LD activity of the patient's serum was inhibited more strongly at 4 degrees C than at 37 degrees C. The decrease of LD activity was more marked in a mixture of the patient's serum with purified LD5 than in that with purified LD1. The immunoglobulin responsible for LD inhibition was an IgA1-lambda. The LD inhibition by the patient's IgA1 was blocked by reduction and alkylation and by NADH. Polymerization of the patient's IgA1 might play an important role in its interaction with LD. Moreover, the possibility exists that part of the patient's IgA1 molecule fits into a pocket of LD in instead of NADH. This is the first report of NADH reversing such LD inhibition. PMID- 17127737 TI - Morphological characterization of the breast in Proteus syndrome complicated by ductal carcinoma in situ. AB - Proteus syndrome (PS) is a severe, variable, and rare disorder with asymmetric and disproportionate overgrowth of body parts, cerebriform connective tissue nevi, epidermal nevi, dysregulated adipose tissue, and vascular malformations. It is associated with benign and occasionally malignant tumors. We report the first case of ductal carcinoma in situ (DCIS) in a 28-yr-old woman with PS who underwent a mastectomy for asymmetric overgrowth. The cut surface of the tissue showed a discrete, white, lobulated, solid mass with multiple cysts with occasional small polypoid nodules. Microscopically, the tissue was characterized by neoplastic and non-neoplastic changes. The former consisted of multiple intraductal papillomas and low-grade intraductal papillary, solid, and cribriform carcinoma. The non-neoplastic changes were characterized by cysts of various sizes, lined by cuboidal or apocrine cells, focally with epithelial papillary proliferation; the lumens contained eosinophilic, mucicarmine-positive, and PAS positive material. Variable ductal proliferation and periductal, peri- and intra lobular fibrosis with loose fibrous connective tissue was present. The carcinoma was positive for ER, PR, CK7, and MIB-1 (40%), and negative for p53 and CK20 staining. We conclude that DCIS may be one of the tumors associated with PS and that the proliferative phenotype serves as an initiator for carcinogenesis. This case highlights the difficulty of recognizing small foci of carcinoma in an asymmetrical overgrowth of the breast in a young woman with PS. PMID- 17127738 TI - Extramedullary hematopoiesis in breast after neoadjuvant chemotherapy for breast carcinoma. AB - We report incidental extramedullary hematopoiesis (EMH) in breasts of 2 patients following neoadjuvant chemotherapy for locally advanced breast cancer. Neither of the patients had a history of hematologic disorders. After chemotherapy, one of the patients had a complete pathologic response and the other had residual carcinoma. In both cases, EMH was mostly seen as myelopoiesis in a background of chemotherapy-induced changes. In the patient with residual carcinoma, EMH was observed in the contralateral prophylactic mastectomy specimen. EMH should be considered a diagnostic pitfall in the differential diagnosis of unusual cellular infiltrates in breast after neoadjuvant chemotherapy. To our knowledge, the association of EMH and neoadjuvant chemotherapy has not been previously reported. PMID- 17127739 TI - Polyclonal plasma cell proliferation with marked hypergammaglobulinemia and multiple autoantibodies. AB - A 77-yr-old man presented with marked peripheral blood and bone marrow plasmacytosis, marked hypergammaglobulinemia, and multiple autoantibodies. Serum protein immunofixation and immunophenotyping of bone marrow plasma cells by flow cytometry and immunohistochemistry disclosed polyclonal proliferation of plasma cells at various stages of differentiation. The presence of multiple autoantibodies in the patient's serum suggests that an autoimmune disease underlies the polyclonal proliferation of plasma cells. PMID- 17127740 TI - The link between koilocytes and human papillomaviruses. PMID- 17127741 TI - Alternative RNA splicing regulation in the testis. AB - Alternative splicing regulation has been shown to be critically important for several developmental pathways. It is particularly prevalent in the testis, which is the site of an extensive adult developmental programme. Alternative splicing is controlled by a splicing code, in which transcripts respond to subtle cell type-specific variations in positive and negative trans-acting RNA-binding proteins according to their unique set of binding sites for these proteins. Because of their unique combinations of cis-acting sequence elements, specific transcripts are able to respond individually to this code. In this review, we discuss how this code may be deciphered in germ cells to mediate a splicing response. PMID- 17127742 TI - Immunocontraception of mammalian wildlife: ecological and immunogenetic issues. AB - Immunocontraception involves stimulating immune responses against gametes or reproductive hormones thus preventing conception. The method is being developed for the humane control of pest and overabundant populations of mammalian wildlife. This paper examines three fundamental issues associated with its use: (1) the difficulties of obtaining responses to self-antigens, (2) the likely evolution of genetically based non-response to immunocontraceptive agents, and (3) the possible changes in the array of pathogens possessed by the target species after generations of immunocontraception. Our review of the literature demonstrates that the barriers to an effective immunocontraceptive are at present very basic. Should they be overcome, the effects of immunocontraception on the immunogenetic constitution of wildlife populations through the selection for non responders must be examined. We suggest that the attempt to use the animal's own immune system to modulate reproduction may be incompatible with the basic biological function of protection against infectious disease. Research programs on mammalian immunocontraception should involve measurement of the heritability of non-response and an assessment of the likely change in the response of the contracepted population to possible pathogens. PMID- 17127743 TI - Fluorescence resonance energy transfer analysis of mitochondrial:lipid association in the porcine oocyte. AB - The role of endogenous lipid in the provision of energy during in vitro maturation of immature porcine oocytes has been studied. Fluorescence resonance energy transfer (FRET) acceptor bleaching methods have been used to examine mitochondrial:lipid droplet co-localisation in live oocytes. FRET experiments demonstrate whether organelles are within the FRET-distance (i.e. 6-10 nm), thus showing true association on a molecular scale. Immature and in vitro-matured porcine oocytes were stained with Mitotracker Green (MTG; mitochondria) and Nile Red (NR; lipid droplets). The data indicated sufficient overlap between MTG emission and NR excitation to support a FRET reaction and that mitochondria and lipid droplets were sufficiently co-localised for a FRET reaction to occur. When NR-stained lipid droplets were specifically bleached, a significant increase in the MTG signal in stained mitochondria was observed (FRET efficiency, E=22.2 +/- 3.18%). These results strongly suggest a metabolic role for lipid metabolism during oocyte maturation. This conclusion was reinforced by the use of inhibitors of fatty acid beta-oxidation, methyl palmoxirate or mercaptoacetate, exposure to which during oocyte maturation led to developmental failure post-fertilisation. These data provide strong evidence that MTG and NR can act as a FRET pair and that in porcine oocytes, mitochondria and lipid droplets lie within 6-10 nm of each other, indicating association on a molecular scale. The findings also suggest that endogenous triglycerides play an important role in energy metabolism during porcine in vitro maturation. PMID- 17127744 TI - Early zygotes are suitable recipients for bovine somatic nuclear transfer and result in cloned offspring. AB - Cloning by somatic cell nuclear transfer (SCNT) subverts sperm-mediated fertilization that normally leads to physiological activation of the oocyte. Therefore, artificial activation is required and it is presently unclear what developmental consequences this has. In this study, we aimed to improve cattle cloning efficiency by utilizing a more physiological method of activating SCNT reconstructs. We carried out in vitro fertilization (IVF) of zona-intact bovine oocytes before SCNT. We removed the zona pellucida 4 h after insemination, stained the fertilized eggs with Hoechst 33342 and mechanically removed both male and female chromatin. The enucleated pre-activated cytoplasts were fused with male adult ear skin fibroblasts ("IVF-NT" group). Chemically activated SCNT embryos, produced according to our standard operating procedure for zona-free SCNT, served as controls. After 7 days, in vitro development to blastocysts of morphological grade 1-3 or grade 1-2 was very similar in both groups (39 vs 40% and 20 vs 21% respectively). However, post-implantation development was improved after sperm-mediated activation. Across four replicate runs, pregnancy establishment at day 35 was significantly higher for IVF-NT than for control SCNT embryos (30/49 = 61 vs 17/41 = 42% respectively; P < 0.05). Development into calves at term or weaning was also higher in the IVF-NT group compared with control SCNT (9/49 = 18 vs 3/41 = 7% and 6/49 = 12 vs 3/41 = 7%; P = 0.11 and 0.34 respectively). PMID- 17127745 TI - Developmental ability of cloned embryos from neural stem cells. AB - The success rate is generally higher when cloning mice from embryonic stem (ES) cell nuclei than from somatic cell nuclei, suggesting that the embryonic nature or the undifferentiated state of the donor cell increases cloning efficiency. We assessed the developmental ability of cloned embryos derived from cultured neural stem cell (NSC) nuclei and compared the success rate with that of embryos cloned from other donor cells such as differentiated NSCs, cumulus cells, Sertoli cells and ES cells in the mouse. The transfer of two-cell cloned embryos derived from cultured NSC nuclei into surrogate mothers produced five live cloned mice. However, the success rate (0.5%) was higher in embryos cloned from cultured NSC nuclei than from differentiated NSCs (0%), but lower than that obtained by cloning mice from other cell nuclei (2.2-3.5%). Although the in vitro developmental potential to the two-cell stage of the cloned embryos derived from NSC nuclei (73%) was similar to that of the cloned embryos derived from other somatic cell nuclei (e.g., 85% in Sertoli cells and 75% in cumulus cells), the developmental rate to the morula-blastocyst stage was only 7%. This rate is remarkably lower than that produced from other somatic cells (e.g., 50% in Sertoli cells and 54% in cumulus cells). These results indicate that the undifferentiated state of neural cells does not enhance the cloning efficiency in mice and that the arrest point for in vitro development of cloned embryos depends on the donor cell type. PMID- 17127746 TI - Proline-rich tyrosine kinase2 is involved in F-actin organization during in vitro maturation of rat oocyte. AB - Microfilaments (actin filaments) regulate various dynamic events during meiotic maturation. Relatively, little is known about the regulation of microfilament organization in mammalian oocytes. Proline-rich tyrosine kinase2 (Pyk2), a protein tyrosine kinase related to focal adhesion kinase (FAK) is essential in actin filaments organization. The present study was to examine the expression and localization of Pyk2, and in particular, its function during rat oocyte maturation. For the first time, by using Western blot and confocal laser scanning microscopy, we detected the expression of Pyk2 in rat oocytes and found that Pyk2 and Try402 phospho-Pyk2 were localized uniformly at the cell cortex and surrounded the germinal vesicle (GV) or the condensed chromosomes at the GV stage or after GV breakdown. At the metaphase and the beginning of anaphase, Pyk2 distributed asymmetrically both in the ooplasm and the cortex with a marked staining associated with the chromosomes and the region overlying the meiotic spindle. At telophase, Pyk2 was observed in the cleavage furrows in addition to its cortex and cytoplasm localization. The dynamics of Pyk2 were similar to that of F-actin, and this kinase was found to co-localize with microfilaments in several developmental stages during rat oocyte maturation. Microinjection of Pyk2 antibody demolished the microfilaments assembly and also inhibited the first polar body (PB1) emission. These findings suggest an important role of Pyk2 for rat oocyte maturation by regulating the organization of actin filaments. PMID- 17127747 TI - NuMA distribution and microtubule configuration in rabbit oocytes and cloned embryos. AB - The assembly of microtubules and the distribution of NuMA were analyzed in rabbit oocytes and early cloned embryos. Alpha-tubulin was localized around the periphery of the germinal vesicle (GV). After germinal vesicle breakdown (GVBD), multi-arrayed microtubules were found tightly associated with the condensed chromosomes and assembled into spindles. After the enucleated oocyte was fused with a fibroblast, microtubules were observed around the introduced nucleus in most reconstructed embryos and formed a transient spindle 2-4 h post-fusion (hpf). A mass of microtubules surrounded the swollen pseudo-pronucleus 5 hpf and a normal spindle was formed 13 hpf in cloned embryos. NuMAwas detected in the nucleus in germinal vesicle-stage oocytes, and it was concentrated at the spindle poles in both meiotic and mitotic metaphase. In both donor cell nucleus and enucleated oocyte cytoplasm, NuMA was not detected, while NuMA reappeared in pseudo-pronucleus as reconstructed embryo development proceeded. However, no evident NuMA staining was observed in the poles of transient spindle and first mitotic spindle in nuclear transfer eggs. These results indicate that NuMA localization and its spindle pole tethering function are different during rabbit oocyte meiosis and cloned embryo mitosis. PMID- 17127748 TI - Interactions between progesterone and tumor necrosis factor-alpha in the regulation of primordial follicle assembly. AB - Follicle assembly is the process by which groups or "nests" of oocytes break down to form primordial follicles. The size of the primordial follicle pool is the major determinant of the reproductive lifespan of a female. Previously, progesterone (P(4)) has been shown to inhibit follicle assembly, while tumor necrosis factor-alpha (TNFalpha) has been shown to promote the apoptosis that is necessary for follicle assembly. The present study examines how TNFalpha and progesterone interact to regulate primordial follicle assembly. Ovaries were collected from newborn rats and placed in organ culture to examine the actions of P(4) and TNFalpha. P(4) was found to decrease primordial follicle assembly and increase the percentage of unassembled oocytes both in vitro and in vivo. TNFalpha treatment did not change the proportion of assembled follicles in cultured ovaries, but blocked the ability of P(4) to inhibit follicle assembly. Microarray analysis of the ovarian transcriptome revealed that progesterone treatment of the ovaries altered the expression of 513 genes with 132 only expressed after P(4) treatment and 16 only expressed in control ovaries. The majority of genes were up-regulated greater than twofold over control, with a small subset of 16 genes down-regulated. Categories of genes affected by P(4) are described including a group of extracellular signaling factors. The progesterone receptors expressed at the time of follicle assembly included the surface membrane progesterone receptors PGRMC1, PGRMC2, and RDA288. The nuclear genomic P(4) receptor was not expressed at appreciable levels. Progesterone increased the expression of several genes (TANK, NFkappaB, Bcl2l1, and Bcl2l2) involved in a signaling pathway that promotes cell survival and inhibits apoptosis. Observations indicate that P(4) acts through the surface membrane progesterone receptors to regulate primordial follicle assembly, and that TNFalpha can override the inhibitory actions of P(4) on follicle assembly. A major mechanism involved in the actions of P(4) is an increase in cell survival genes and inhibition of the apoptosis pathway. Observations provide insight into the hormonal regulation of primordial follicle assembly and lead to novel approaches to potentially manipulate follicle assembly and reproductive capacity. PMID- 17127749 TI - Expression of the adaptor protein m-Numb in mouse male germ cells. AB - Numb is an adaptor protein that is asymmetrically inherited at mitosis and controls the fate of sibling cells in different species. The role of m-Numb (mammalian Numb) as an important cell fate-determining factor has extensively been described mostly in neural tissues, particularly in progenitor cells, in the mouse. Biochemical and genetic analyses have shown that Numb acts as an inhibitor of the Notch signaling pathway, an evolutionarily conserved pathway involved in the control of cell proliferation, differentiation, and apoptosis. In the present study, we sought to determine m-Numb distribution in germ cells in the postnatal mouse testis. We show that all four m-Numb isoforms are widely expressed during postnatal testis development. By reverse transcriptase-PCR and western blot analyses, we further identify p71 as the predominantly expressed isoform in germ cells. Moreover, we demonstrate through co-immunoprecipitation studies that m Numb physically associates with Ap2a1, a component of the endocytotic clathrin coated vesicles. Finally, we employed confocal immunofluorescence microscopy of whole mount seminiferous tubules and isolated germ cells to gain more insight into the subcellular localization of m-Numb. These morphological analyses confirmed m-Numb and Ap2a1 co-localization. However, we did not observe asymmetric localization of m-Numb neither in mitotic spermatogonial stem cells nor in more differentiated spermatogonial cells, suggesting that spermatogonial stem cell fate in the mouse does not rely on asymmetric partitioning of m-Numb. PMID- 17127750 TI - Analysis of furin ectodomain shedding in epididymal fluid of mammals: demonstration that shedding of furin occurs in vivo. AB - Sperm cell surface proteins and proteins of their surrounding fluids are reported to be proteolytically processed in relation to acquisition of sperm fertility during epididymal transit. Several of these proteins might be potential targets for subtilisin-like pro-protein convertase. Using immunochemistry and mass spectrometry analysis, we found that an 80 kDa form of furin (EC 3.4.21.75) is present in the fluid from the mid-caput to the distal corpus regions of the epididymis of various domestic mammals. This protein is absent from the fluid of the caudal region, suggesting that it is reabsorbed or degraded. The cDNA sequence of ovine furin was obtained and the mRNA was found throughout this organ, although in greater amounts in the mid and distal caput regions. Metabolic labeling with (35)S-amino acids indicated that the protein was synthesized and released from the epithelium only in a restricted area of the mid-caput, suggesting a specific regionalized mechanism of secretion. The fluid protein is not pelleted at 100 000 g and did not react with a C-terminal antibody indicating that it is not bound to membranous materials. These findings demonstrate that a furin ectodomain shedding occurs naturally in vivo in the epididymis where this enzyme could be involved in fluid and/or sperm membrane protein processing. PMID- 17127751 TI - Control of oestradiol secretion and of cytochrome P450 aromatase messenger ribonucleic acid accumulation by FSH involves different intracellular pathways in oestrogenic bovine granulosa cells in vitro. AB - The objective of this study was to determine the major intracellular signalling pathways used by FSH and insulin to stimulate cytochrome P450 aromatase (Cyp19) mRNA and oestradiol accumulation in oestrogenic bovine granulosa cells in vitro. Bovine granulosa cells from small follicles (2-4 mm diameter) were cultured for 6 days under non-luteinizing conditions in the presence of insulin at 100 ng/ml, or insulin (10 ng/ml) and FSH (1 ng/ml). On day 4 of culture, specific inhibitors of phosphatidylinositol 3-kinase (PI3K; LY-294002), protein kinase C (PKC; GF 109203X), protein kinase A (PKA; H-89) or mitogen-activated protein (MAP) kinase activation (PD-98059) were added. The addition of PI3K and PKC inhibitors, but not of PKA inhibitor, significantly decreased insulin-stimulated Cyp19 mRNA levels and oestradiol accumulation (P < 0.001). The PKA inhibitor significantly decreased FSH-stimulated Cyp19 mRNA abundance and oestradiol secretion, whereas PI3K and PKC inhibitors decreased oestradiol secretion without affecting Cyp19 mRNA accumulation. Inhibition of MAP kinase pathway significantly increased Cyp19 mRNA abundance in insulin- and FSH-stimulated cells. P450scc mRNA levels and progesterone secretion were not affected by any inhibitor in either experiment. Although FSH stimulates Cyp19 expression predominantly through PKA, oestradiol secretion is altered by PI3K and PKC pathways independently of Cyp19 mRNA levels. In addition, we suggest that Cyp19 is under tonic inhibition mediated through a MAP kinase pathway. PMID- 17127752 TI - Secreted phosphoprotein 1 (osteopontin) is expressed by stromal macrophages in cyclic and pregnant endometrium of mice, but is induced by estrogen in luminal epithelium during conceptus attachment for implantation. AB - Secreted phosphoprotein 1 (SPP1, osteopontin) is the most highly upregulated extracellular matrix/adhesion molecule/cytokine in the receptive phase human uterus, and Spp1 null mice manifest decreased pregnancy rates during mid gestation as compared with wild-type counterparts. We hypothesize that Spp1 is required for proliferation, migration, survival, adhesion, and remodeling of cells at the conceptus-maternal interface. Our objective was to define the temporal/spatial distribution and steroid regulation of Spp1 in mouse uterus during estrous cycle and early gestation. In situ hybridization localized Spp1 to luminal epithelium (LE) and immune cells. LE expression was prominent at proestrus, decreased by estrus, and was nearly undetectable at diestrus. During pregnancy, Spp1 mRNA was not detected in LE until day 4.5 (day 1 = vaginal plug). Spp1-expressing immune cells were scattered within the endometrial stroma throughout the estrous cycle and early pregnancy. Immunoreactive Spp1 was prominent at the apical LE surface by day 4.5 of pregnancy and Spp1 protein was also co-localized with subsets of CD45-positive (leukocytes) and F4/80-positive (macrophages) cells. In ovariectomized mice, estrogen, but not progesterone, induced Spp1 mRNA, whereas estrogen plus progesterone did not induce Spp1 in LE. These results establish that estrogen regulates Spp1 in mouse LE and are the first to identify macrophages that produce Spp1 within the peri-implantation endometrium of any species. We suggest that Spp1 at the apical surface of LE provides a mechanism to bridge conceptus to LE during implantation, and that Spp1 positive macrophages within the stroma may be involved in uterine remodeling for conceptus invasion. PMID- 17127753 TI - Reduced nitric oxide synthase and cyclo-oxygenase activity in the uterus of non obese diabetic mice. AB - A functional interaction between progesterone, Th2 cytokines and a suitable balance between nitric oxide and prostaglandins in the uterus is considered to have a major role in the success of embryo implantation and pregnancy. Non-obese diabetic (NOD) mice offer a suitable model to study the modulatory role of Th1 cytokines on uterus signalling and function, since at the prediabetic stage they develop a spontaneous Th1 autoimmune response against exocrine glands similar to Sjogren's syndrome. Vasoactive intestinal peptide (VIP) is a vasoactive neuro- and immunopeptide that promotes Th2 profiles and contributes to the smooth muscle relaxation and vasodilation. The aim of the present study was to investigate the activities of nitric oxide synthase and cyclo-oxygenase and the effect of VIP in the uterus of NOD mice with an emerging Th1 cytokine response. We present evidence of a reduced basal and VIP-stimulated activity of both enzymes in the uterus of NOD mice compared with normal BALB/c mice in proestrus. An altered functional interaction between both enzymes is also present in NOD mice at the time when increased levels of serum interleukin (IL)-12 and tumour necrosis factor-alpha but not interferon (IFN)-gamma or IL-10 were detected. We conclude that signalling alterations in uteri of NOD mice are simultaneous to the onset of a systemic Th1 cytokine response. PMID- 17127754 TI - Porcine endometrial and conceptus tissue kallikrein 1, 4, 11, and 14 gene expression. AB - Previous studies have suggested that the porcine endometrium may express several tissue kallikreins during the estrous cycle and early pregnancy. The present study investigated porcine endometrial and conceptus tissue kallikrein 1, 4, 11, and 14 mRNA expression during the estrous cycle and early pregnancy. Tissue kallikrein (KLK) gene expression was evaluated using quantitative RT-PCR and in situ hybridization. KLK1 expression was similar across the estrous cycle and early pregnancy, and localized to the endometrial luminal (L) and glandular (G) epithelium. KLK4 endometrial mRNA expression was greatest on days 0, 5, and 10 when compared with days 12, 15, and 17 of the estrous cycle and greater in cyclic compared with pregnant gilts. Expression of KLK4 was more intense in the stroma and uterine epithelium from days 0 to 10 of the estrous cycle. Endometrial KLK11 mRNA was not different between cyclic and pregnant gilts but the expression was greatest on days 10 and 12 compared with all other days evaluated. There was an increased intensity of KLK11 gene expression in the stratum compactum on day 10 of the estrous cycle and early pregnancy. Endometrial KLK14 mRNA expression was not detectable on days 5 and 10 but was expressed on days 0, 12, 15, and 17 of the estrous cycle and pregnancy. KLK14 expression was localized in the uterine L and G epithelium, and stroma throughout the endometrium after day 10. Conceptus KLK1 mRNA did not change from days 10 to 17 of gestation. However, conceptus KLK4, and 14 mRNA expression was greatest on day 10 with expression declining after day 14 of gestation. Expression of the various tissue kallikreins in the endometrium and conceptus during the estrous cycle and early pregnancy in the pig can serve in the activation of growth factors and tissue remodeling during the establishment of pregnancy. PMID- 17127755 TI - Relationships between sperm DNA fragmentation, sperm apoptotic markers and serum levels of CB-153 and p,p'-DDE in European and Inuit populations. AB - Persistent organochlorine pollutants (POPs) are suspected to interfere with hormone activity and the normal homeostasis of spermatogenesis. We investigated the relationships between sperm DNA fragmentation, apoptotic markers identified on ejaculated spermatozoa and POP levels in the blood of 652 adult males (200 Inuits from Greenland, 166 Swedish, 134 Polish and 152 Ukrainian). Serum levels of 2, 2', 4, 4', 5, 5'-hexachlorobiphenyl (CB-153), as a proxy of the total POP burden, and of 1,1-dichloro-2,2-bis(p-chlorophenyl)-ethylene (p,p'-DDE), as a proxy of the total DDT exposure were determined. Sperm DNA fragmentation was measured by using the TUNEL assay, whereas immunofluorescence methods were utilized for detecting pro-apoptotic (Fas) and anti-apoptotic (Bcl-xL) markers. Both TUNEL assay and apoptotic markers were statistically differed across the four populations. No correlation between neither sperm DNA fragmentation nor apoptotic sperm parameters and the large variations in POPs exposure was observed for the separate study groups. However, considering the European populations taken together, we showed that both %TUNEL positivity and Bcl-xL were related to CB-153 serum levels, whereas our study failed to demonstrate any relations between DDE and %TUNEL positivity and apoptotic sperm biomarkers (Fas and Bcl-xL) in any region or overall regions. These results suggest that CB-153 and related chemicals might alter sperm DNA integrity and Bcl-xL levels in European adult males, but not in the highly exposed Inuit men. Additional issues (genetic background, lifestyle habits and characterization of total xeno-hormonal activities) need to be investigated in order to fully assess the population variations observed. PMID- 17127756 TI - Comparative trial of the canine parvovirus, canine distemper virus and canine adenovirus type 2 fractions of two commercially available modified live vaccines. AB - The results of vaccinating two groups of puppies with commercial vaccines, both of which claimed to provide adequate protection with a final vaccination at 10 weeks of age, were compared. Groups of 19 and 20 puppies with similar titres of maternally derived antibodies against canine parvovirus (cpv), canine distemper virus (cdv) and canine adenovirus type 2 (cav-2) at four weeks of age were vaccinated at six and 10 weeks of age and their responses to each vaccination were measured by comparing the titres against cpv, cdv and cav-2 in the serum samples taken immediately before the vaccination and four weeks later. After the vaccination at six weeks of age, all 19 of the puppies in group 1 had responded to cpv and cdv, and 14 had responded to cav-2; in group 2, 17 of the 20 had responded to cpv, 19 to cdv and 15 to cav-2. In both groups the puppies that did not respond to the first vaccination had responded serologically to cpv, cdv and cav-2 at 10 weeks of age. PMID- 17127757 TI - Survey of pigs' kidneys with lesions consistent with PMWS and PDNS and ochratoxicosis. Part 1: concentrations and prevalence of ochratoxin A. AB - One thousand condemned pigs' kidneys were collected in February 2002 from two pig abattoirs in England to assess the possible contribution of ochratoxicosis to postweaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS); 250 of the kidneys with macroscopic lesions consistent with nephrosis/nephritis (pale or white cortical lesions) were selected, and the concentration of ochratoxin A was measured in samples of renal cortex by high-performance liquid chromatography (HPLC). Low concentrations were detected in 230 (92 per cent) of the kidneys tested, and in 41 (16.4 per cent) of them the concentration was below the limit of quantification of 0.2 microg/kg. In 187 (74.8 per cent) of the kidneys, the concentration was more than 0.2 microg/kg, and the highest concentration detected was 2.3 microg/kg. The mean (sd) concentration was 0.31 (0.33) microg/kg. The identification of ochratoxin A was confirmed by mass spectrometry. The concentrations of ochratoxin A did not exceed the threshold assessed by the Food Standards Agency to be safe for human food. PMID- 17127758 TI - Field evaluation of a PCR for the diagnosis of chlamydial abortion in sheep. AB - A total of 94 vaginal swab samples and 195 serum samples collected from aborted ewes in 15 flocks were examined by pcr and a complement fixation test, respectively. In addition, 172 samples of stomach contents from fetuses from different flocks submitted for the diagnosis of abortion during the four lambing periods between 2000 and 2004 were tested by pcr. Chlamydial dna was detected in seven vaginal swabs obtained from five of the 15 flocks and in six samples of fetal stomach contents. The results of pcr and flock serology for Chlamydia were positive in five of the 15 flocks and negative in eight. PMID- 17127759 TI - Molecular typing of transmissible spongiform encephalopathy from Italian disease outbreaks in small ruminants. PMID- 17127760 TI - Efficacy and safety of selamectin in the treatment of Otodectes cynotis infestation in domestic ferrets. PMID- 17127761 TI - Plasma cortisol levels in dogs undergoing diagnostic imaging procedures. PMID- 17127762 TI - Ultrasonographic findings in three cows with ureteral obstruction due to urolithiasis. PMID- 17127763 TI - Description of the first case of equine infectious anaemia in Northern Ireland. PMID- 17127765 TI - Communication during suspected notifiable disease incidents. PMID- 17127766 TI - Unusual findings in a llama. PMID- 17127767 TI - FMD and the contiguous cull. PMID- 17127768 TI - Equine interspecies aggression. PMID- 17127770 TI - Bacterial flavodoxins support nitric oxide production by Bacillus subtilis nitric oxide synthase. AB - Unlike animal nitric-oxide synthases (NOSs), the bacterial NOS enzymes have no attached flavoprotein domain to reduce their heme and so must rely on unknown bacterial proteins for electrons. We tested the ability of two Bacillus subtilis flavodoxins (YkuN and YkuP) to support catalysis by purified B. subtilis NOS (bsNOS). When an NADPH-utilizing bacterial flavodoxin reductase (FLDR) was added to reduce YkuP or YkuN, both supported NO synthesis from either L-arginine or N hydroxyarginine and supported a linear nitrite accumulation over a 30-min reaction period. Rates of nitrite production were directly dependent on the ratio of YkuN or YkuP to bsNOS. However, the V/Km value for YkuN (5.2 x 10(5)) was about 20 times greater than that of YkuP (2.6 x 10(4)), indicating YkuN is more efficient in supporting bsNOS catalysis. YkuN that was either photo-reduced or prereduced by FLDR transferred an electron to the bsNOS ferric heme at rates similar to those measured for heme reduction in the animal NOSs. YkuN supported a similar NO synthesis activity by a different bacterial NOS (Deinococcus radiodurans) but not by any of the three mammalian NOS oxygenase domains nor by an insect NOS oxygenase domain. Our results establish YkuN as a kinetically competent redox partner for bsNOS and suggest that FLDR/flavodoxin proteins could function physiologically to support catalysis by bacterial NOSs. PMID- 17127771 TI - Novel n-3 fatty acid oxidation products activate Nrf2 by destabilizing the association between Keap1 and Cullin3. AB - Consumption of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) can mitigate the progression of diseases in which oxidative stress represents a common underlying biochemical process. Nrf2-regulated gene expression regulates detoxification of reactive oxygen species. EPA and DHA were subjected to an in vitro free radical oxidation process that models in vivo conditions. Oxidized n-3 fatty acids reacted directly with the negative regulator of Nrf2, Keap1, initiating Keap1 dissociation with Cullin3, thereby inducing Nrf2-directed gene expression. Liquid chromatography-tandem mass spectrometry analyses of oxidized EPA demonstrated the presence of novel cyclopentenone-containing molecules termed J3-isoprostanes in vitro and in vivo and were shown to induce Nrf2-directed gene expression. These experiments provide a biochemical basis for the hypothesis that formation of J-ring compounds generated from oxidation of EPA and DHA in vivo can reach concentrations high enough to induce Nrf2-based cellular defense systems. PMID- 17127772 TI - Hu proteins regulate polyadenylation by blocking sites containing U-rich sequences. AB - A recent genome-wide bioinformatic analysis indicated that 54% of human genes undergo alternative polyadenylation. Although it is clear that differential selection of poly(A) sites can alter gene expression, resulting in significant biological consequences, the mechanisms that regulate polyadenylation are poorly understood. Here we report that the neuron-specific members of a family of RNA binding proteins, Hu proteins, known to regulate mRNA stability and translation in the cytoplasm, play an important role in polyadenylation regulation. Hu proteins are homologs of the Drosophila embryonic lethal abnormal visual protein and contain three RNA recognition motifs. Using an in vitro polyadenylation assay with HeLa cell nuclear extract and recombinant Hu proteins, we have shown that Hu proteins selectively block both cleavage and poly(A) addition at sites containing U-rich sequences. Hu proteins have no effect on poly(A) sites that do not contain U-rich sequences or sites in which the U-rich sequences are mutated. All three RNA recognition motifs of Hu proteins are required for this activity. Overexpression of HuR in HeLa cells also blocks polyadenylation at a poly(A) signal that contains U-rich sequences. Hu proteins block the interaction between the polyadenylation cleavage stimulation factor 64-kDa subunit and RNA most likely through direct interaction with poly(A) cleavage stimulation factor 64-kDa subunit and cleavage and polyadenylation specificity factor 160-kDa subunit. These studies identify a novel group of mammalian polyadenylation regulators. Furthermore, they define a previously unknown nuclear function of Hu proteins. PMID- 17127773 TI - Cholesterol import fails to prevent catalyst-based inhibition of ergosterol synthesis and cell proliferation of Trypanosoma brucei. AB - Trypanosoma brucei (TB) cultured in rat blood, bovine serum, or lipid-depleted serum generated distinct differences in cholesterol availability. Whereas cell proliferation of the parasite was relatively unaffected by cholesterol availability, the ratios of cellular ergostenols to cholesterol varied from close to unity to 3 orders of magnitude different with cholesterol as the major sterol (>99%) of bloodstream form cells. In the procyclic form cultured with lipid depleted serum, 15 sterols at 52 fg/cell were identified by GC-MS. The structures of these sterols reveal a nonconventional ergosterol pathway consistent with the novel product diversity catalyzed by the recently cloned sterol methyltransferase (SMT). A potent transition state analog of the TB SMT C24 alkylation reaction, 25 azalanosterol (25-AL; inhibition constant Ki = 39 nM), was found to inhibit the growth of the procyclic and bloodstream forms at an IC(50) of approximately 1 microM. This previously unrecognized catalyst-specific inhibition of cell growth was unmasked further using the 25-AL-treated procyclic form, which, compared with control cultures, caused a change in cellular sterol content from ergostenols to cholesterol. However, growth of the bloodstream form disrupted by 25-AL was not rescued by cholesterol absorption from the host, suggesting an essential role for ergosterol (24-methyl sterol) in cell proliferation and that the SMT can be a new enzyme target for drug design. PMID- 17127774 TI - Utility of esmolol in thyroid crisis. AB - Thyroid storm is an uncommon but potentially life-threatening manifestation of hyperthyroidism. Mortality can be 30-60% in hospitalized patients unless appropriately treated by combined therapy. We report a case of a 25-year-old African American woman with past medical history of Graves disease and moderately persistent asthma who presented to the emergency department with signs and symptoms of thyrotoxic crisis. Therapy instituted and included the use of an esmolol infusion for control of hypersympathetic activity. A review of the clinical presentation, diagnosis, and management of thyrotoxic crisis is presented along with a discussion on the choice of beta blockade therapy. PMID- 17127775 TI - Pain response to M-M-R vaccination in 4-6 year old children. AB - BACKGROUND: Differences in pain response to two different M-M-R products have previously been demonstrated in 12-month old infants and in 4 a 6 year old children. Objective To determine if the acute and immediate pain response to two licensed M-M-R vaccine products (using a self-report measure) in children 4-6 years of age was similar to that demonstrated in younger infants. METHODS: Randomized, double blind, study. Subjects were randomly allocated to PriorixA (SmithKline Beecham) or M-M-R IIA (Merck Frosst). The primary outcome measure was pain response to vaccination quantified using a self-report OUCHER pain scale. Secondary outcome measures included pain measurement by proxy (physician and parent) using a visual analog scale (VAS) and measurement of cry and cry duration immediately post-vaccination. RESULTS: Of the 60 subjects enrolled, 30 received PriorixA and 30 received M-M-R IIA. There were no significant differences between the two groups on age, sex, or previous painful procedure. Post-vaccination, children in the M-M-R IIA group had higher median pain scores compared with children in the PriorixA group for VAS (12.5 vs. 2.0, respectively by paediatricians, p=0.017; 18.5 vs. 5.0, respectively by parents, p=0.235), OUCHER (20 vs. 0.00, respectively, p=0.047). The median duration of crying post M-M-R IIA was higher compared with PriorixA (6 vs. 0 seconds, respectively, p=0.020). Conclusion PriorixA was associated with significantly less pain compared with M-M R IIA, at the time of injection. PMID- 17127776 TI - Comparative study of intranasal midazolam and intravenous diazepam sedation for procedures and seizures. AB - OBJECTIVE: To evaluate the safety and efficacy of intranasal midazolam for seizures and various procedures. METHODS: Prospective randomized study. Total 125 children of all ages of either sex, for seizure episode (n-76) and various invasive and non-invasive procedures (n-49) received either intranasal midazolam (0.2 mg/Kg) or intravenous diazepam (0.3 mg/Kg). RESULTS. Mean time from arrival at hospital to starting treatment was significantly shorter in midazolam group compared to diazepam group [2.34+/-0.90; minute vs 4.61+/-1.08 minute p< 0.001]. Mean time to control seizures after arrival in hospital was significantly shorter in midazolam group compared to diazepam group [5.25+/-0.86 minute vs 6.51+/-1.06 minute p. CONCLUSION: Midazolam by the intranasal route provides safe and equally effective non-invasive method of sedation for procedures and seizures. PMID- 17127777 TI - Randomized controlled trial of ipratropium bromide and salbutamol versus salbutamol alone in children with acute exacerbation of asthma. AB - OBJECTIVE: To evaluate effect of addition of ipratropium to salbutamol delivered by metered dose inhaler and spacer in the beginning of treatment of mild to moderate exacerbation of asthma. METHODS: Children between 5 to 15 years of age with mild to moderate exacerbation of asthma were randomized to receive either a combination of ipratropium bromide and salbutamol or salbutamol alone administered by metered dose inhaler and spacer. The effects on clinical asthma score and spirometric parameters were compared. RESULTS: A total of 60 children were randomized in the study. The baseline characteristics of two groups were comparable. Children getting combination of salbutamol and ipratropium showed significantly greater improvement in percent-predicted PEFR and FEF25-75% than children receiving salbutamol alone. CONCLUSION: There was beneficial effect of addition of ipratropium to salbutamol administered by MDI with spacer at the beginning of therapy for mild to moderate acute exacerbation of asthma in children. PMID- 17127778 TI - Red cell pyruvate kinase deficiency in neonatal jaundice cases in India. AB - OBJECTIVE: Pyruvate Kinase (PK) deficiency is the most common enzymopathy of the glycolytic pathway in erythrocytes. It constitutes one of the common causes of hereditary non-spherocytic hemolytic anemia. The aim of this study was to screen newborns in India for pyruvate kinase (PK) deficiency in relation to unconjugated hyperbilirubinemia. METHODS: Laboratory investigations done included complete blood counts, reticulocyte counts, direct and indirect bilirubin, assay of G6PD and PK activity, ATP and 2,3 DPG levels. All variables were studied in 50-cord blood samples from normal deliveries and 218 neonates with hyperbilirubinemia. RESULTS: 7 of the 218 cases of neonatal jaundice were PK deficient with 30-40% reduction in PK activity. These cases also had a 3-4-fold increase in 2,3 DPG:ATP ratios, which is one of the additional indicators for PK deficiency. Six of the 7 infants had a severe clinical course. CONCLUSION: This study shows that the prevalence of PK deficiency in Indian neonatal jaundice cases is 3.21%, which is relatively high. This emphasizes the need for screening neonatal hyperbilirubinemia cases in India for PK deficiency. PMID- 17127779 TI - Mantoux and contact positivity in tuberculosis. AB - OBJECTIVE: To study the role of Mantoux and contact history in various forms of Childhood tuberculosis. METHODS: 605 children registered with TB clinic of Institute of Child Health and Hospital for Children, Chennai over a 5 year period from January 2000 to October 2005 with various forms of tuberculosis were recruited in the study. Clinical examination findings, basic investigations, chest skiagrams, computerized tomography (CT) wherever warranted, sputum or gastric aspirates for AFB smear, histopathology wherever possible were analyzed. RESULTS: The study showed that Mantoux positivity in various forms of tuberculosis studied is 34.7%. The positivity of Mantoux was highest in lymph node tuberculosis (53%) and the lowest with CNS tuberculosis (21.2%). Among the other forms, Mantoux positivity was 36.4% in TB abdomen, 44.4% in Skeletal TB, 30.3% in pulmonary tuberculosis. The contact positivity was 30.4% in the sample studied. CONCLUSION: The study also reflects that the extra pulmonary forms of tuberculosis seems to be more common in the pediatric population which constituted 79.8% of the cases included in the study. PMID- 17127780 TI - Maternal deprivation, acute respiratory infections and immune regulation. AB - OBJECTIVE: The study was designed to investigate the acute respiratory disease incidence (DI) in deprived infants and test the hypothesis if maternal deprivation effects the processes of immunoregulation in infants. METHODS: The prospective study during 1 year in Tbilisi Infant's House was performed. Cohort of 136 infants at age from 1 to 24 months without any congenital abnormalities formed the basic group. The cohort of 136 healthy infants at age 1 to 24 months of the same population living under the maternal care in three shelters of Tbilisi region was randomized as a control. The study included: DI, age of first attack, duration of illness, outcome, plasma immune parameters. RESULTS: The data have demonstrated that maternal deprivation induces a marked increase in the severity of acute respiratory disease among infants. DI was twice as much in deprived children as in control group. Moreover, there was diagnosed the disruption of normal correlations between plasma CD3, CD4 and CD8 in infants under maternal deprivation. CONCLUSION: Maternal deprivation induces changes in processes of immunoregulation in infants resulted in elevation of acute respiratory DI among them. PMID- 17127781 TI - Occult neurovesical dysfunction with anorectal malformations. AB - OBJECTIVE: To evaluate for the occurrence of occult NVD in children with anorectal malformations (ARM) using urodynamic evaluation. METHODS: This prospective study was carried out on children with ARM prior to and following definitive procedure. Urodynamic studies were performed on the Phoenix Griffon machine (Albyn Medical) using Phoenix plus software. RESULT: Nineteen children in the age range of 3 months to 156 months (mean = 19.2) were included in this study. Among these 19 children 13 underwent re-evaluation after definitive surgery for ARM. There were 11(57.9%) males and 8(42.1%) females. Of the 19 children 14 (73.7%) were cases of high anorectal malformation (HARM) and 5 (26.3%) were cases of low anorectal malformation (LARM). Baseline evaluation done in 19 children revealed seven urodynamic patterns: Normal capacity, compliant without uninhibited contractions (UIC) (21.1%); Normal capacity, compliant with UIC (5.3%); Normal capacity, poorly compliant without UIC (5.3%); Normal capacity, poorly compliant with UIC (10.5%); small capacity, compliant with UIC (5.3%); Small capacity, poorly compliant with UIC (26.3%) and large capacity, complaint with UIC (26.3%). Thirteen patients were evaluated post operatively also and in only 23% (3 of 13) no change in urodynamic pattern were observed. In the remaining 76.9% (10 of 13) some changes in urodynamics pattern were observed. The deleterious changes observed were appearance of UIC in 30.8% (4 of 13), decrease in the bladder capacity in 23% (3 of 13) and decrease in bladder compliance in 15.4% (2 of 13). CONCLUSION: Only 9 of of the 19 patients had normal urodynamics pre-operatively and post-operatively 3 more patients worsened. Incidence of occult NVD is high in patients with ARM even in the absence of clinical and radiological evidence of vertebral or lower urinary tract abnormalities. Though there seems to be a high incidence of changes in the neurovesical functions of these patients following definitive corrective surgery for ARM only time will show whether this has any deleterious effect on the upper tracts. PMID- 17127783 TI - Developmentally supportive care and NIDCAP. AB - With increasing advances in field of perinatal medicine and new breakthroughs in technology, the perinatal mortality has shown a steady decline worldwide. This has given rise to a new dimension, that of developmentally oriented care of these sick neonates while they are still in intensive care. NIDCAP &developmentally supportive care is aimed at decreasing the stress levels in these babies by individualized care which also involves the family. Growth and development has been shown to get enhanced by this approach. There are only few well designed studies on this issue of extreme importance. This article presents a brief review of current literature as well as outlines relevant policies followed in the author's unit. PMID- 17127784 TI - Addressing the "New" NEC: Part I: rediscovering the basics. AB - Epithelial cell functions ultimately define the ability of the extremely low birth weight human fetus to survive outside of the uterus. These specialized epithelial cell capacities manage all human interactions with the ex utero world including: (i) lung mechanics, surface chemistry and gas exchange, (ii) renal tubular balance of fluid and electrolytes, (iii) barrier functions of the intestine and skin for keeping bacteria out and water in, plus enabling intestinal digestion, as well as (iv) maintaining an intact neuroepithelium lining of the ventricles of the brain and retina. In Part I of this two part review, the authors describe why the gut barrier is a clinically relevant model system for studying the complex interplay between innate and adaptive immunity, dendritic &epithelial cell interactions, intraepithelial lymphocytes, M-cells, as well as the gut associated lymphoid tissues where colonization after birth, clinician feeding practices, use of antibiotics as well as exposure to prebiotics, probiotics and maternal vaginal flora all program the neonate for a life-time of immune competence distinguishing "self" from foreign antigens. These barrier defense capacities become destructive during disease processes like necrotizing enterocolitis (NEC) when an otherwise maturationally normal, yet dysregulated and immature, immune defense system is associated with high levels of certain inflammatory mediators like TNFa. In Part II, the authors will discuss the theoretical advantages of using rhG-CSF in managing NEC or sepsis by augmenting neonatal neutrophil number and killing capacity including an unexpected, paradoxical and potent anti-TNFa function that may serve to limit extension of tissue destruction without impairing bacterial killing capacity. The authors conclude by arguing that NEC may be the ideal disease process for testing whether a clearly defined clinical benefit of cytokine therapy can prove beneficial. PMID- 17127785 TI - Toward improving mucosal barrier defenses: rhG-CSF plus IgG antibody. AB - Epithelial cell functions ultimately define the ability of the extremely low birth weight human fetus to survive outside of the uterus. These specialized epithelial cell capacities manage all human interactions with the ex utero world including: (i) lung mechanics, surface chemistry and gas exchange, (ii) renal tubular balance of fluid and electrolytes, (iii) barrier functions of the intestine and skin for keeping bacteria out and water in, plus enabling intestinal digestion, as well as (iv) maintaining an intact neuroepithelium lining of the ventricles of the brain and retina. In Part I of this two part review, the authors describe why the gut barrier is a clinically relevant model system for studying the complex interplay between innate and adaptive immunity, dendritic &epithelial cell interactions, intraepithelial lymphocytes, M-cells, as well as the gut associated lymphoid tissues where colonization after birth, clinician feeding practices, use of antibiotics as well as exposure to prebiotics, probiotics and maternal vaginal flora all program the neonate for a life-time of immune competence distinguishing "self" from foreign antigens. These barrier defense capacities become destructive during disease processes like necrotizing enterocolitis (NEC) when an otherwise maturationally normal, yet dysregulated and immature, immune defense system is associated with high levels of certain inflammatory mediators like TNFa. In Part II the authors discuss the rationale for why rhG-CSF has theoretical advantages in managing NEC or sepsis by augmenting neonatal neutrophil number, neutrophil expression of Fcg and complement receptors, as well as phagocytic function and oxidative burst. rhG-CSF also has potent anti-TNFa functions that may serve to limit extension of tissue destruction while not impairing bacterial killing capacity. Healthy, non-infected neutropenic and septic neonates differ in their ability to respond to rhG-CSF; however, no neonatal clinical trials to date have identified a clear clinical benefit of rhG-CSF therapy. This manuscript will review the literature and evidence available for identifying the ideal subject for cytokine treatment using NEC as the model disease target. PMID- 17127786 TI - Prevention of bronchopulmonary dysplasia: finally, something that works. AB - Due to a lack of understanding of the molecular mechanisms involved in its pathogenesis, bronchopulmonary dysplasia (BPD) still remains a major cause of morbidity and mortality in the premature infant and there is no effective preventive and/or therapeutic intervention. We have taken a basic biologic approach to elucidate the pathophysiology of BPD and have discovered that disruption of the alveolar Parathyroid Hormone-related Protein (PTHrP) signaling is centrally involved in this process. Further, stabilization of this signaling pathway by using exogenous PTHrP agonists can prevent and/or rescue the molecular injuries caused by insults that lead to BPD. Based upon years of work in this field, here I provide a novel and innovative molecular approach, i.e, exogenous treatment with PTHrP pathway agonists to prevent and/or treat BPD. However, to avoid any later surprises, it is important to emphasize that before translating it into human trials, this approach needs further testing and refinement in animal models. PMID- 17127787 TI - Acute myeloid leukemia with severe aplastic anemia following immunosuppressive therapy. AB - Severe aplastic anemia (AA) is a life-threatening condition wherein bone marrow transplantation (BMT) is the therapy of choice in a young patient who has a matched sibling donor. Here, we report an 11-year-old boy with severe AA who was referred for BMT late in its course when he had developed acute myeloid leukemia following two courses of immunosuppressive therapy with antithymocyte globulin and cyclosporin. He was then treated with induction therapy using cytosine arabinoside and daunomycin for acute myeloid leukemia, but he succumbed due to infection and refractory leukemia. We discuss the relevance of early referral for BMT in severe AA. PMID- 17127788 TI - Imaging findings in hepatic Langerhans' cell histiocytosis. AB - We describe ultrasonographic and computed tomographic features of hepatic lesions in two cases of disseminated Langerhans' cell histiocytosis affecting children. In the first case, hyperechoic band like periportal lesions were observed at ultrasonography, which on computed tomography was found to be hypodense admixed with fatty attenuation (HU@23 to - 57) at places. In addition, the caudate lobe was very prominent. In the second case, the hepatic parenchyma showed predominantly hyperechoic diffusely heterogeneous echogenicity. There were features of cirrhosis of liver with portal hypertension in the form of atrophy of right lobe with hypertrophy of left lobe of liver with lobulated outline, prominent main portal vein and splenoportal axis, splenomegaly and gastroesophageal varices. Both the patients were put on chemotherapy as per schedule (Protocol: DAL HX - 83) and are on follow up. PMID- 17127789 TI - Rare serotype non-typhoidal Salmonella sepsis. AB - An 11-month female with a poor socio-economic status presented to a tertiary care paediatric hospital with complaints of fever of 4-5 days and diarrhoea of 20 days duration. The patient didn't respond to the prescribed antimicrobials namely- Norfloxacin and metronidazole. On admission she was diagnosed as persistent diarrhea with PEM grade III with sepsis. Stool examination and culture were negative for any pathogens, however blood culture yielded growth of Salmonella Virchow which was susceptible to most common antimicrobial agents excepting Trimethoprim Sulfamethoxazole. Salmonella Virchow is a common non-typhoidal Salmonellae causing bacteremia in the west, however this is the first report of bacteremia by S. virchow from India. PMID- 17127791 TI - Pediatric trauma--social awareness and rehabilitation measures. PMID- 17127790 TI - Twins with senior-Loken syndrome. AB - Senior-Loken syndrome is a rare entity that combines familial juvenile nephronophthisis with retinal dystrophy. The eye disease may be congenital amaurosis of Leber type or pigmentary retinal degeneration and electroretinogram (ERG) helps in the diagnosis of these varieties. The disease progresses inexorably to chronic renal failure. Here is a case of twins with Senior-Loken syndrome emphasizing the importance of ophthalmic examination in children with renal failure, for determining a correct diagnosis. PMID- 17127792 TI - Effectiveness of individualized education program for slow learners. PMID- 17127793 TI - Relation of obesity to acute myocardial infarction in Japanese patients. AB - BACKGROUND: It remains uncertain whether obesity is an independent risk factor for coronary heart disease in young adults, as well as adults, in Japan. METHODS AND RESULTS: In the present study, 1,260 cases of acute myocardial infarction (AMI) and 3,775 community controls were recruited from the AMI-Kyoto Multi-Center Risk Study and Kyoto Citizen's Health and Nutrition Study, respectively. Obesity and other risk factors were retrospectively examined between cases and controls in each subgroup of young males (20-40 years), middle-aged males or females (40 60 years), older males or females (60-80 years), and very old males or females (80-100 years). In young, middle-aged, and older males, as well as in older females, cases had a higher body mass index (BMI) than controls. In young males, as well as in middle-aged and older females, cases had a higher prevalence of smoking than controls. Except for very old males, the prevalences of hypercholesterolemia, hypertension, and diabetes mellitus were higher in each subgroup of cases than in controls. Multivariate logistic regression analysis revealed that obesity (BMI >or=25) was an independent risk for AMI in young and middle-aged males, but not in females, whereas smoking was an independent risk for AMI in middle-aged and older females as well as in older males. CONCLUSIONS: Obesity is significantly associated with AMI, independent of the classic coronary risk factors, in young and middle-aged males. These findings support the current emphasis on controlling obesity to prevent coronary events in young Japanese male adults. PMID- 17127794 TI - Prognostic value of the metabolic syndrome for long-term outcomes in patients undergoing percutaneous coronary intervention. AB - BACKGROUND: The prevalence of metabolic syndrome (MS), regarded as an important risk factor for coronary artery disease, is growing. However, the relationship between MS and long-term outcomes after percutaneous coronary intervention (PCI) in the Japanese patient population remains unknown. METHODS AND RESULTS: Seven hundred and forty-eight consecutive patients who underwent PCI were assessed. Patients were categorized by the presence or absence of MS using the NCEP-ATPIII definition (for obesity, a body mass index >or=25 kg/m(2) was used). Kaplan-Meier estimation and Cox proportional hazards model were used for unadjusted and adjusted analyses for all cause mortality and cardiac events. The progress of 318 (42.5%) patients with MS and 430 (57.5%) patients without MS was analyzed. The mean follow-up was 12.0+/-3.6 years. Overall, there were 88 (11.8%) deaths from all causes, and there were no significant differences between the 2 groups. The occurrence of cardiac events was significantly higher in the MS group than that in the no MS group (25.5% vs 15.6%, hazard ratio 2.23; 95% confidence interval 1.59-3.11; p<0.001). CONCLUSIONS: The presence of MS significantly increased the risks of subsequent cardiac events among patients who underwent PCI. PMID- 17127795 TI - Impact of PercuSurge device conjugative with intracoronary administration of nitroprusside on no-reflow phenomenon following primary percutaneous coronary intervention. AB - BACKGROUND: The present study tested the hypothesis that when administered in conjunction with a PercuSurge device for treatment of acute myocardial infarction (AMI), intracoronary (IC) administration of nitroprusside (NTP) is safe and superior to IC administration of NTP alone or nitroglycerin (NTG) for reversing slow-flow or no-reflow, both of which occur frequently during primary percutaneous coronary intervention (PCI). METHODS AND RESULTS: Sixty-two patients with ST-segment elevation AMI of <12 h duration undergoing primary PCI were enrolled. When the final Thrombolysis In Myocardial Infarction (TIMI) flow was normal (TIMI-3), NTG 200 microg was administered first, followed by (5 min later) NTP 100 microg via an intra-guiding catheter. When final TIMI flow was 10 years. Fifteen (13%) of 114 patients who had follow-up echocardiography became ;end-stage' HCM and 8 patients died of severe and refractory heart failure. They already showed minimally dilated left ventricular (LV) dimension and lower LV fractional shortening at initial evaluation. CONCLUSIONS: Although HCM patients in a Japanese rural community showed relatively benign clinical course (the same as cohorts in the developed world), heart failure death because of LV remodeling became equally important to sudden death when they were followed for more than 10 years. PMID- 17127797 TI - Impact of ethanol dosing on the long-term outcome of alcohol septal ablation for obstructive hypertrophic cardiomyopathy: a single-center prospective, and randomized study. AB - BACKGROUND: The impact of ethanol dose on the long-term outcome of alcohol septal ablation (ASA) for obstructive hypertrophic cardiomyopathy was investigated. METHODS AND RESULTS: Fifty-four patients (age 24-82 years; 65% women) undergoing ASA were randomized into 2 groups according to the dose of injected ethanol: Group A 1-2 ml, Group B >2 ml. Clinical and echocardiographic data were obtained at baseline and during follow-up. The volume of ethanol injected was 1.50+/-0.4 and 2.60+/-0.6 ml (p<0.001) with a subsequent peak of creatine kinase-MB of 2.25+/-1.00 and 2.62+/-1.57 microkat/L (p=0.02) in Groups A and B, respectively. The median follow-up was 39 (range 6-72) months after ASA, during which 1 patient died and 1 repeat procedure was necessary in both groups of patients. Both groups had a significant and similar improvement in outflow pressure gradient, dyspnea (New York Heart Association functional class) and angina pectoris (Canadian Cardiovascular Society class) (p<0.001). There was a significant decrease in the left ventricular ejection fraction (LVEF) in Group B (81+/-7 vs 75+/-7%; p=0.002), but not in Group A (80+/-7 vs 79+/-7%; p=0.67). Thinning of the basal septum was more pronounced in Group B than in Group A (9.3+/-5.7 vs 6.6+/-3.4 mm; p=0.04). CONCLUSIONS: A lower dose of ethanol injected into the target septal branch reduces both the size of necrosis and subsequent thinning of the basal septum, and preserves LVEF during long-term follow-up. Moreover, the low dose (1 2 ml) is as safe and as hemodynamically efficacious as higher doses. PMID- 17127798 TI - Temporal worsening of sleep-disordered breathing in the acute phase of myocardial infarction. AB - BACKGROUND: Sleep apnea is an important risk factor for cardiovascular diseases, but whether the severity of sleep-disordered breathing (SDB) changes in the acute phase of myocardial infarction (MI) has not been well determined, nor has it been determined what type of SDB, central or obstructive, (CSA or OSA) is exacerbated. METHODS AND RESULTS: Polysomnography was performed in patients with acute phase of MI during the acute (days 3-5) and chronic (day 14) phases. On the same day, the ventilatory equivalent (VE)/carbon dioxide production (VCO(2)) slope, urinary catecholamines secretion and arterial carbon dioxide tension were assessed before sleep. The apnea/hypopnea index was significantly decreased in the chronic phase (13.26+/-11.30 vs 6.97+/-5.67, p<0.05). The distribution of the types of SDB was unchanged, indicating both CSA and OSA can be exacerbated in the acute phase of MI. The VE/VCO(2) slope and arterial carbon dioxide tension before sleep were also unchanged. Urinary norepinephrine secretion was slightly decreased, although the difference was not significant. CONCLUSIONS: SDB is temporarily worsened in the acute phase of AMI and both CSA and OSA are worsened in AMI. PMID- 17127799 TI - Association of hyperadiponectinemia with severity of ventricular dysfunction in congestive heart failure. AB - BACKGROUND: Adiponectin, which is a collagen-like plasma protein produced by adipose tissue, has anti-atherogenic and anti-inflammatory effects. Plasma adiponectin levels in patients with congestive heart failure (CHF) were determined, as well as relationships between the plasma levels of adiponectin and other hormones. METHODS AND RESULTS: The study group comprised 90 patients with CHF and 20 control subjects, who were divided into 4 subgroups according to New York Heart Association (NYHA) functional class. Plasma levels of adiponectin, tumor necrosis factor (TNF)-alpha and brain natriuretic peptide (BNP) and cardiac hemodynamics were determined. Plasma adiponectin levels were significantly increased according to the severity of NYHA class in the patients with CHF; control: 6.2+/-1.0; NYHA I: 8.5+/-1.9, NYHA II: 12.0+/-2.2, NYHA III: 13.0+/-2.7, NYHA IV: 14.9+/-2.7 microg/ml (p=0.0008). Similarly, plasma BNP levels were significantly increased in accordance with the NYHA class. Plasma adiponectin levels correlated positively with BNP (r=0.40, p=0.0002) and TNF-alpha (r=0.49, p=0.0001), and correlated negatively with cardiac index (r=-0.27, p=0.05). In 24 of 46 patients in the NYHA III and IV subgroups, according to the prompt improvement in cardiac function, levels of both plasma adiponectin and BNP were significantly reduced (p<0.0001). CONCLUSION: Plasma adiponectin levels increased according to the severity of CHF and, moreover, they correlated with the plasma levels of BNP and TNF-alpha. These results indicate that augmented release of adiponectin is involved in the pathogenesis of CHF and further study is needed to elucidate its exact role. PMID- 17127800 TI - Functional mitral regurgitation during exercise in patients with heart failure. AB - BACKGROUND: Functional mitral regurgitation (MR) is common in patients with chronic heart failure (CHF) and left ventricular dysfunction. The severity of CHF is expressed in terms of exercise tolerance, so MR during exercise would affect the severity of heart failure. However, it is not well known how much MR increases during exercise, or if it is related with severity of heart failure. METHODS AND RESULTS: Seventeen subjects underwent dynamic cycle exercise at steady-state levels of 80% and 150% of anaerobic threshold (AT). During each exercise level, the MR jet and left atrial (LA) area were measured, and the degree of MR was expressed as the ratio (MR/LA). The MR/LA increased slightly at 80% AT (rest: 15.5+/-7.8%, 80% AT: 21.7+/-9.3%, p<0.05). It increased more at 150% AT (29.2+/-11.6%, p<0.01). The MR/LA at both 80% and 150% AT weakly correlated with peak oxygen consumption/heart rate (r=-0.509). They showed a weakly positive correlation with the ventilatoly equivalent/CO(2) production slope (r=0.340). CONCLUSIONS: MR during exercise increases as stroke volume worsens during exercise, so evaluation of MR during exercise is important in the management of patients with CHF. PMID- 17127801 TI - Portable-type signal-averaged electrocardiography with dipyridamole to detect patients with coronary artery disease. AB - BACKGROUND: In a retrospective study portable-type signal-averaged electrocardiography (SAECG) with dipyridamole stress was found to identify patients with coronary artery disease (CAD) at their bedside with high sensitivity and specificity, so the utility of this method was prospectively investigated in the present study. METHODS AND RESULTS: Standard 12-lead QRS wave SAECG was performed before and after dipyridamole stress at the bedside in 71 patients with chest pain (43 males, mean age 63 +/-9 years). The filtered QRS duration (fQRSd) before and after dipyridamole stress was determined by multiphasic oscillation method for each of the standard 12 leads, and the maximal value of changes in fQRSd (MAX DeltafQRSd) among the 12 leads was determined. The positive test was defined as MAX DeltafQRSd >or=5 ms, and negative as MAX DeltafQRSd <5 ms based on the previous study. Selective coronary arteriography was performed next. In the positive group (n=31), 25 patients had significant stenosis of the coronary artery and 6 did not. In the negative group (n=40), 5 patients had significant stenosis and 35 did not. The sensitivity, specificity, positive predictive accuracy and negative predictive accuracy for CAD detection by SAECG was 83%, 85%, 81% and 88%, respectively. CONCLUSIONS: Dipyridamole stress portable SAECG is useful for detecting CAD at the patient's bedside with high sensitivity and specificity. PMID- 17127802 TI - Atrial electrophysiological abnormality in patients with Brugada syndrome assessed by P-wave signal-averaged ECG and programmed atrial stimulation. AB - BACKGROUND: There is evidence that some patients with Brugada syndrome (BS) exhibit atrial tachyarrhythmias including paroxysmal atrial fibrillation. We investigated whether BS associated not only with vulnerability to ventricular fibrillation, but also with vulnerability to atrial fibrillation. METHODS AND RESULTS: In 15 patients with BS and Brugada-type electrocardiogram (ECG) (14 men, 1 woman; age 52.8+/-12.9 years) and 15 age-matched control patients (12 men, 3 women; age 50.9+/-18.9 years), the P-wave signal-averaged ECG was recorded, and the filtered P-wave duration was derived from the vector magnitude obtained by X, Y, Z leads. In 11 of the 15 patients with BS and Brugada-type ECG, invasive electrophysiologic testing was conducted. Filtered P-wave duration was significantly increased in patients with BS and Brugada-type ECG in comparison with control subjects (143.2+/-12.9 vs 129.6+/-10.1 ms, p<0.001). Ventricular late potential (root mean square voltage <20 muV in the last 40 ms and <40 muV at a low amplitude signal duration >38 ms) was present in 10 of the 12 BS patients in whom a QRS wave signal-averaged electrogram was also recorded. In all 11 patients with Brugada-type ECG who underwent electrophysiologic testing, sustained atrial fibrillation (>5 min) was induced by 1 or 2 atrial extrastimuli. In 10 of these 11 patients, ventricular fibrillation was also induced by 2 or 3 right ventricular extrastimuli. CONCLUSIONS: The electrical abnormality in BS is not limited to the ventricular level; similar changes occur in the atria. Such abnormal conduction properties could be a substrate for re-entrant atrial tachyarrhythmias. PMID- 17127803 TI - Clinical and electrophysiological characteristics of binodal disease. AB - BACKGROUND: Although coexistence of atrioventricular conduction disturbances with sick sinus syndrome (SSS), so-called binodal disease (BND), is a frequently encountered disorder, its clinical significance and electrophysiological characteristics remain unknown. METHODS AND RESULTS: One hundred and seven patients with SSS were divided into BND (n=30) and N-BND groups (n=77). Sinus cycle length, sinus node recovery time (SRT), sino-atrial conduction time (SACT), the number of isolated sinus node electrograms, atrio-His (AH) interval, His ventricular (HV) interval, intra-atrial conduction time (PA intervals) and QRS width were measured. In addition, the prevalence of bundle-branch block was obtained. The parameters of sino-atrial and intra-atrial conduction were significantly longer in the BND group: SRT (5,070+/-2,628 vs 3,122+/-1,856 ms, p<0.05), SACT (115+/-30 vs 87+/-21 ms, p<0.05), PA intervals (56+/-13 vs 41+/-8 ms, p<0.05). The BND group was more likely to have atrial fibrillation than the N BND group (83.3% vs 53.2%, p<0.01). HV interval, QRS width and the prevalence of associated bundle-branch block did not differ between the 2 groups. CONCLUSION: BND patients not only had sino-atrial and atrioventricular node dysfunction, but also widespread atrial conduction disturbances. Thus, in the clinical setting BND should be categorized as severe SSS. PMID- 17127804 TI - Usefulness of rapid low-dose/high-dose 1-day 99mTc-sestamibi ECG-gated myocardial perfusion single-photon emission computed tomography. AB - BACKGROUND: The clinical usefulness of a rapid rest low-dose/stress high-dose (dose ratio =1:5) (99m)Tc-sestamibi myocardial perfusion single-photon emission computed tomography (SPECT) protocol for the detection of coronary artery disease was evaluated. METHODS AND RESULTS: In 89 patients, rest images were obtained immediately after the injection of (99m)Tc-sestamibi (256.1+/-28.4 MBq) followed by drinking water (400 ml). Exercise or vasodilator stress test was performed immediately after the completion of rest imaging with the injection of (99m)Tc sestamibi (1312.3 +/-167.6 MBq). Prior to the post-stress imaging, patients were asked again to drink water (400 ml) in order to eliminate subdiaphragmatic tracer activity. The myocardial count ratio (stress/rest) of (99m)Tc-sestamibi was calculated. Image quality was scored using a 4-point scale system (4= excellent, 3= good, 2= poor, 1= unacceptable). Coronary angiography was performed in 56 patients within 1 month of the SPECT scan. All patients successfully performed the protocol and total examination time was 108+/-7 min. The myocardial count ratio of (99m)Tc-sestamibi was always greater than 6. The image quality was satisfactory both at rest (3.4+/-0.9) and after stress (3.9+/-0.2). The sensitivity and specificity to detect coronary artery stenosis >50% was 84% and 97%, respectively. CONCLUSIONS: This rapid one-day (99m)Tc-sestamibi protocol provides adequate image quality and diagnostic accuracy for detecting coronary artery disease. PMID- 17127805 TI - Multidetector row computed tomography noninvasively assesses coronary reperfusion after thrombolytic therapy in patients with ST elevation myocardial infarction. AB - BACKGROUND: The study objective was to assess the efficacy of 16-slice multidetector row computed tomography (MDCT) in estimating residual stenosis and successful reperfusion after thrombolysis in patients with ST-elevation myocardial infarction (STEMI). METHODS AND RESULTS: A total of 31 patients with STEMI underwent MDCT scanning within 6 h (mean 4.6+/-1.1) after thrombolysis and the results for detection of significant residual stenosis and distal flow of the infarct-related artery were compared with those from conventional coronary angiography (CCAG) performed within 24 h (mean 12.1+/-5.6) after the MDCT scan. Successful reperfusion was defined as Thrombolysis In Myocardial Infarction flow 2 or 3 on CCAG and full contrast enhancement of the distal artery landmarks on MDCT. A final analysis was performed using 24 patients (312 segments). MDCT had a positive predictive value of 73.3% and a negative predictive value of 95.1% for detecting significant residual stenosis. It accurately estimated 17 of 18 patients (94.4%) with successful reperfusion and 5 of 6 (83.3%) with failed reperfusion on the basis of comparison with CCAG. CONCLUSIONS: MDCT demonstrated high accuracy not only for the detecting residual stenosis, but also for assessing successful reperfusion after thrombolytic therapy in patients with STEMI. PMID- 17127806 TI - Young adult patients with short dissection length and thrombosed false lumen without ulcer-like projections are liable to have false-negative results of D dimer testing for acute aortic dissection based on a study of 113 cases. AB - BACKGROUND: Previous reports indicate that D-dimer testing (DT) for acute aortic dissection (AAD) has a sensitivity of 100%, but each study comprised less than 30 patients. The aim of this study was to evaluate the positive rate and factors related to the results of DT for AAD in a larger population. METHODS AND RESULTS: DT (cutoff; upper normal limit) was performed for 113 consecutive AAD patients within 24 h of symptom onset. In total, 104 (92%) patients exhibited positive DT. The positive rate of DT showed a low tendency in patients aged less than 70 years and for a time interval from symptom onset to admission within 120 min, and there were significant differences between those with and without a thrombosed false lumen (TFL) (86.4% (n=59) vs 98.1% (n=54), p=0.033), complete TFL (excluding patients with ulcer-like projection (ULP) from those with a TFL) (81.1% (n=37) vs 97.4% (n=76), p=0.005) and length score (1 (n=28); 78.6%, 2 (n=40); 95.0%, 3 (n=45); 97.8%, p=0.005). Multivariate analysis demonstrated age (odds ratio =1.164, p=0.013), complete TFL (0.048, 0.030) and length score (6.271, 0.033) as independent factors. CONCLUSIONS: Physicians should be aware that younger patients with short dissection length and a TFL without ULP are liable to have false-negative DT results. PMID- 17127807 TI - Patients with low high-density lipoprotein-cholesterol or smoking are more likely to develop myocardial infarction among subjects with a visible lesion or stenosis in coronary artery. AB - BACKGROUND: Traditional contrast coronary arteriography affords only an indirect view of aspects of atheromata related to their propensity to trigger thromboses, so it is urgent to recognize the vulnerable person who is more likely to develop myocardial infarction (MI) among patients with visible lesion or stenosis in coronary artery. METHODS AND RESULTS: Two hundred and eighty-eight patients (144 MI patients, 144 controls) who had either a visible lesion or differing extent of stenosis in 1 or more major coronary arteries were consecutively enrolled. Lipid profile, C-reactive protein (CRP), smoking, hypertension, dyslipidemia and diabetes were analyzed for their association with MI. No differences in the prevalence of dyslipidemia, hypertension or diabetes was found between the patients with MI and those without, and CRP, triglycerides, total cholesterol and low-density lipoprotein-cholesterol levels did not differ between the 2 groups (all p>0.05). However, high-density lipoprotein-cholesterol (HDL-C) was significantly lower in the patients with MI than in those without (1.06+/-0.30 vs 1.14+/-0.32 mmol/L, p=0.024). On multivariate analysis after adjustment for age and gender, adjusted odds ratio (95% confidence interval) of MI was 0.44 (0.20 0.96) for HDL-C, p=0.038; 2.6 (1.48-4.56, p=0.001) for smoking, which indicated that high HDL-C was protective for MI, and smoking was associated with an increased risk of MI. CONCLUSIONS: The present findings indicate that among subjects with a visible lesion or stenosis in coronary arteries, those with low HDL-C or smokers are more likely to develop MI. PMID- 17127808 TI - Apolipoprotein E polymorphism influences lipid phenotypes in Chinese families with familial combined hyperlipidemia. AB - BACKGROUND: Apolipoprotein E (apoE) polymorphism is associated with changes in the lipoprotein profile of individuals with familial combined hyperlipidemia (FCHL), but its effects on the lipoprotein profiles of members of Chinese families with FCHL remain uncertain. METHODS AND RESULTS: 43 FCHL families (n=449) and 9 normolipidemic families (n=73) were recruited to assess the influence of apoE polymorphism on plasma lipids. The relative frequency of the epsilon4 allele in affected and unaffected FCHL relatives, spouses and normolipidemic members was 13.8%, 5.3%, 9.1% and 6.8%, respectively, with a significantly higher frequency in affected FCHL relatives, compared with unaffected FCHL relatives or normolipidemic members (p=0.0002 or p=0.029). In FCHL relatives, the apoE4 subset (E4/4 and E4/3) exhibited significantly higher levels of apoB, total cholesterol and low-density lipoprotein-cholesterol (LDL-C) than did the apoE3 (E3/3) subset, especially in women (all p<0.05), and there was significant elevation of LDL-C concentrations in men only (p<0.05). In men, the apoE2 (E3/2) subset indicated a decreased level of apoB and increased apoA1 compared with those in the apoE3 subset (p<0.05). CONCLUSIONS: ApoE polymorphism appears to be associated with variance of the lipoprotein phenotype in Chinese families with FCHL. PMID- 17127809 TI - Prognostic role of alveolar-arterial oxygen pressure difference in acute pulmonary embolism. AB - BACKGROUND: This study investigated the utility of the alveolar - arterial oxygen pressure difference (AaDO (2)) in predicting the short-term prognosis of acute pulmonary embolism (PE). METHODS AND RESULTS: This study retrospectively enrolled 114 consecutive patients with acute PE, diagnosed by either spiral computed tomography or high probability ventilation - perfusion lung scans. During the first 24 h of admission, all patients had initial artery blood gas collected under room air. Patient exclusion criteria were chronic lung disease, septic emboli, and moderate and low probability lung scans. Patients were assigned to 2 groups based on either 30-day death or a 30-day composite event. Receiver operating characteristic analyses was used to determine the AaDO(2) cut-off value for predicting primary and composite endpoints. Statistical analysis demonstrated significant differences in AaDO(2) between the 30-day composite endpoint group and the 30-day composite event-free survival group (p=0.012). The AaDO(2) had a strong trend between the 30-day death group and the survival group (p=0.062). The best cut-off value for AaDO(2) was 53 mmHg and using this, the positive predictive value for 30-day death was 25% and the negative predictive value was 92%. For the 30-day composite endpoint, the positive predictive value for AaDO(2) was 35%, and the negative predictive value was 84%. In this study, thrombocytopenia was also an indicator of poor prognosis for patients with acute PE. CONCLUSION: The AaDO(2) measurement is a highly useful and simple measurement for predicting short-term prognosis in patients with acute PE. It has high negative predictive value and moderate positive predictive value for 30-day death and 30-day composite event. Aggressive thrombolytic treatment strategies should be considered for patients with an initial poor prognostic parameter (ie, AaDO(2) >or=53 mmHg). PMID- 17127810 TI - Clinical characteristics and outcome of hospitalized patients with heart failure in Japan. AB - BACKGROUND: Heart failure (HF), defined as a complex clinical syndrome that can result from any structural or functional cardiac disorder that impairs the ability of the ventricle to fill with or eject blood, is a leading cause of mortality and hospitalization for adults older than 65 years in the industrialized countries. The characteristics and outcome of patients with HF have been described by several epidemiological studies and large scale clinical trials, performed mainly in the United States and Europe. Very little information is available on this issue in Japan. METHODS AND RESULTS: The Japanese Cardiac Registry of Heart Failure in Cardiology (JCARE-CARD) is designed to prospectively study the characteristics, treatment, and outcomes of a broad sample of patients hospitalized with HF at teaching hospitals throughout Japan between January 2004 to June 2005 and the outcomes, including death and hospital readmission, will be followed through 2006 (mean follow-up at least 1 year). Participating cardiologists identify patients admitted for worsening of HF symptoms. Demographics, medical history, severity, treatment, and outcome data are collected and entered into a database via secure web browser technology. As of June 2005, baseline data for 2,676 patients with HF have been registered from 164 participating hospitals. CONCLUSIONS: The JCARE-CARD will provide important insights into the management of patients with HF in routine clinical practice in Japan, thus providing the framework for improved management strategies for these patients. PMID- 17127811 TI - Japan assessment of pitavastatin and atorvastatin in acute coronary syndrome (JAPAN-ACS): rationale and design. AB - BACKGROUND: Many trials have shown that 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase inhibitors reduce the incidence of cardiovascular events and mortality. One method of decreasing the incidence of cardiovascular events could be to reduce the progression of coronary atherosclerosis, and a recent study found that atorvastatin can cause coronary plaque to regress. To generalize this finding, using conventional HMG-CoA reductase inhibitors at many Japanese centers, randomized trials of pitavastatin and atorvastatin will be conducted with patients with acute coronary syndrome (ACS). METHODS AND RESULTS: Patients with ACS who have undergone successful percutaneous coronary intervention under intravascular ultrasound guidance will be studied. They will be randomly allocated to pitavastatin or atorvastatin groups and followed up for 8-12 months. The primary endpoint will be the percent change in coronary plaque volume, and secondary endpoints will include absolute changes in coronary plaque volume, serum lipid levels and inflammatory markers. The safety profile will also be evaluated. CONCLUSIONS: This study will examine the ability of HMG-CoA reductase inhibitors to regress coronary plaque in Japanese patients with ACS and the findings should help to improve the prognosis of such patients and clarify the involved mechanisms. PMID- 17127812 TI - Echolucent carotid plaques as a feature in patients with acute coronary syndrome. AB - BACKGROUND: Carotid arterial ultrasound examination may be helpful for screening populations at high risk for acute coronary syndrome (ACS), so the present study was designed to identify the carotid arterial characteristics of patients with ACS. METHODS AND RESULTS: Carotid ultrasound examinations were performed in 172 patients with ACS, 166 patients with stable coronary artery disease (CAD), and 96 control subjects. Common carotid arterial structures were assessed by the intima media thickness (IMT), interadventitial diameter (IAD), lumen diameter (LD), the IMT to LD ratio (IMT/LD), and the plaque burden based on the plaque score. Plaque morphology was assessed by the echogenecity based on the gray-scale median (GSM). IMT, IAD, IMT/LD, and plaque score did not differ between the ACS and stable CAD groups. The GSM in the ACS group was lower (47.5+/-25.3, p<0.001) than in the control (70.1+/-22.5) and stable CAD (73.7+/-23.4) groups. Multiple logistic regression analysis showed that the presence of carotid echolucent plaques (GSM 20. The CS was 2.7 +/- 2.1. There was a correlation between PS and the geriatric indices but no correlation between PS and CS. CONCLUSION: PS is significantly correlated with the geriatric indices but is independent of CS. PS appears to be a good parameter for the assessment of global activity in the elderly subject with LC. PMID- 17127907 TI - [Tobacco use among teachers in a district of the Tunisian Sahel]. AB - INTRODUCTION: The aim of this study was to evaluate the scale of tobacco use among teachers in the district of Kalaa Kebira (a semi-urban region in the Tunisian Sahel). METHODS: The study design was descriptive and cross sectional, employing a self administered questionnaire given to 402 teachers. RESULTS: The response rate was 89%. The average age of respondents was 35.7 7.9 years. The global prevalence of smoking among teachers was 29.3% (men: 51.6%; women: 3.6%). 79% of smokers were addicted to nicotine according to the Fagerstrom test. Half of the teachers were poorly informed about the hazards of smoking and 77.9% of smoking teachers didn't refrain from smoking in front of their pupils. CONCLUSIONS: The teachers' lack of knowledge limits their effectiveness both as a role model and information relay in the war against tobacco. PMID- 17127908 TI - [Factors influencing pulmonary function in Tunisian women aged 45 years and more]. AB - INTRODUCTION: The aim of this study was to analyse the factors influencing pulmonary function and inspiratory muscle strength in healthy Tunisian women aged >or=45 years and in particular to determine the effect of parity. METHODS: A medical questionnaire together with an evaluation of sporting activity score and 2 levels for schooling and socio-economic status was administered. Parity was introduced as numeric, as dichotomous (G1:or=4) and in 3 classes (C1:<2; C2:=3-4; C3: > 4). Plethysmography with measurement of airway conductance and maximal inspiratory pressure was performed. RESULTS: 108 women were included. According to the ascending multiple linear regression, and in decreasing order, the following influencing factors are noted: Age and height, parity, weight and daily activity, schooling level, and finally leisure activity, body mass index, and physical activity. With high parity, and especially in women aged >or=60 years, there was a decrease in inspiratory muscle strength and an obstructive tendency, without associated restrictive component. Parity effects are age independent. CONCLUSIONS: Factors influencing the pulmonary function of healthy Tunisian women aged >or=45 years are multiple. Lung function declines with increasing parity. PMID- 17127909 TI - [Contact tracing in a case of tuberculosis in a health care worker]. AB - INTRODUCTION: Tuberculosis is a public health problem, of which the nosocomial transmission from a health care worker to patients has not been well documented. OBSERVATIONS: A Senegalese surgeon aged 32 was admitted to hospital on account of deterioration in his general health. He was found to have sputum positive tuberculosis and received standard treatment. We report the strategy employed for tracing the contacts of this health care worker. Of a total of 185 members of staff (permanent and temporary) and 91 patients who had been in contact with the index case, 180 (97.3%) and 71 (78%) respectively were screened. Of the 251 subjects screened, 5 staff (2.8%) and 11 patients (15.4%) showed evidence of latent tuberculous infection and 6 were treated. In total 97.3% of staff exposed were screened at the beginning of the study and 63% were reviewed at 3 months as opposed to 78% and 53% for the patients. CONCLUSION: This study shows poor compliance with the visit 3 months after exposure and the need to standardise the procedures in order to improve the screening and follow up of contacts. PMID- 17127910 TI - [Semi-invasive necrotic aspergillosis in a child with cystic fibrosis]. AB - INTRODUCTION: Aspergillosis colonisation in cystic fibrosis (CF), usually due to Aspergillus fumigatus (AF), classically presents as allergic bronchopulmonary aspergillosis. However, aspergillus infection can produce a range of manifestations: from minor colonization to an invasive infection. CASE REPORT: A 14-year-old CF patient, chronically colonized with Staphylococcus aureus and Pseudomonas aeruginosa, presented with acute right-sided chest pain, a moderate fever and no modification of the sputum. The chest X-ray showed a 5 cm round opacity. Laboratory parameters were WBC 24,500 G/l, CRP 27 mg/l, Total IgE 1527 UI/l, Specific Aspergillus fumigatus IgE 31 UI/l. Bronchoscopy revealed external compression of the middle lobe bronchus with mucopurulent secretions coming from apical segment of the lower lobe. Bronchoalveolar lavage revealed the presence of pseudomonas aeruginosa 103 CFA/ml, staph aureus 107 CFA/ml and one colony of AF. Chest CT scan showed a large necrotic mass with an air-fluid level located in the apical segment of the right lower lobe. Initial therapy with itraconazole and corticosteroid was replaced by voriconazole, caspofungin, metronidazole and linezolide. Treatment was well tolerated and after 8 weeks the chest X-ray appearances had returned to normal. CONCLUSIONS: The possible diagnoses and therapeutic options are discussed. Conventional antifungal treatment with amphotericin B is limited by nephrotoxicity. These newer antifungal agents appear to be effective and well-tolerated. PMID- 17127911 TI - [Broncholithiasis]. AB - INTRODUCTION: Broncholithiasis is defined as the presence of calcified material in the bronchial lumen. The aim of our work was to study the clinical, radiological and therapeutic aspects of broncholithiasis. OBSERVATIONS: We report the histories of 6 patients identified over 14 years (1990-2004). They included 4 men and 2 women with an average age of 50 years. A past history of pulmonary tuberculosis was found in 2 cases. The presenting symptom was haemoptysis in 4 patients. Fibreoptic bronchoscopy was performed in all patients and broncholiths were found in two. Thoracic CT scan was performed in 5 patients and in 3 showed hilar and parenchymal calcification, suggesting the diagnosis of broncholithiasis. The diagnosis was confirmed in 2 patients by bronchoscopy, in 2 others by surgical biopsy and in the remaining 2 by the CT appearances. Three patients were operated on: 2 for diagnosis and the third for haemostasis. Pulmonary tuberculosis was discovered in 2 patients, one by culture and the other by surgical biopsy. The outcome was satisfactory in all cases. CONCLUSIONS: Broncholithiasis presents a problem of differential diagnosis from other pulmonary pathologies on account of misleading clinical, endoscopic and radiological features. PMID- 17127912 TI - [Pulmonary MALT lymphoma revealing AIDS]. AB - INTRODUCTION: We report the case of a patient with an isolated pulmonary mucosa associated lymphoid tissue (MALT) lymphoma that revealed an acquired immune deficiency syndrome (AIDS). CASE REPORT: A 30 year old man from Central Africa was admitted to hospital with cough, dyspnoea and general weakness. A diagnosis of HIV infection was made promptly. The thoracic CT scan revealed diffuse bilateral ground glass opacities as well as consolidation of the right upper lobe. After a non-diagnostic endoscopy the diagnosis of a low grade B cell MALT lymphoma (CD20+) was made by lung biopsy and confirmed by the presence of the t(11;18) translocation. No extrathoracic lymphoma was found. Treatment with rituximab and triple anti-retroviral therapy led to a rapid and complete remission that was maintained for 3 years after the diagnosis. CONCLUSION: Pulmonary MALT lymphoma may reveal AIDS. A combination of rituximab and anti retroviral therapy led to complete remission in this patient. PMID- 17127914 TI - [Pleural effusion complicating a totally implanted venous device]. AB - INTRODUCTION: Totally implanted venous devices (TIVD) are an essential tool for repeated intravenous treatments such as chemotherapy for cancer and antibiotics for cystic fibrosis. CASE REPORT: A woman of 76 years was treated for bronchiectasis, colonised by Pseudomonas aeruginosa, with courses of intravenous antibiotics. On account of poor peripheral veins a TIVD was implanted. The implantation and subsequent antibiotic injections were uncomplicated. Three years later a further course of antibiotics was prescribed. As no reflux of blood was obtained on puncturing the device the position of the catheter was checked radiologically. The patient complained of right shoulder pain following each infusion. The chest x-ray showed a pleural effusion. The sudden onset of the effusion, the absence of signs of infection and the patient's good clinical condition suggested pleural extravasation of the antibiotic infusion from the TIVD. Pleural aspiration yielded 1400 mls of watery fluid. Injection of contrast into the TIVD confirmed intra-pleural extravasation. CONCLUSION: When using a TIVD it is important to bear in mind the possibility of late pleural complications. PMID- 17127913 TI - [Severe breathlessness in a patient presenting with chronic bronchitis]. AB - INTRODUCTION: Dyspnoea is the major symptomatic complaint of patients with chronic obstructive pulmonary disease (COPD). Awareness of an unpleasant respiratory sensation, occurs through a complex system of ventilatory control where, as in pain perception, supra-bulbar structures play a major role. In the hyperventilation syndrome, dramatic episodes of dyspnea can occur without any change in peripheral afferent signals and thus do not predictably represent a real deterioration in pulmonary function in patients who also have COPD. CASE REPORT: We report the observation of a patient with GOLD stage 0 COPD who demonstrated a major limitation of daily activities due to supra-bulbar influences on respiratory control which triggered repetitive periods of hyperventilation and generated almost constant dyspnea. CONCLUSIONS: Hyperventilation syndrome can be difficult to diagnose in patients with respiratory disease, although, in most cases, the discrepancy between the severity of symptomatic complaint compared to the degree of alteration of lung function in combination with specific tests may suggest the diagnosis. PMID- 17127915 TI - [The pseudoalveolar form of sarcoidosis: a diagnostic pitfall]. AB - BACKGROUND: Sarcoidosis is a benign systemic granulomatosis whose aetiology remains unknown. Lung is the most frequently involved organ. The pseudoalveolar form of this disease is known to have an acute onset and is quite uncommon. Therefore, diagnosing such a rare variety of sarcoidosis is rather often challenging. OBSERVATIONS: In the present article, the authors report two cases of pseudoalveolar sarcoidosis. The patients, both young adults, showed no suggestive signs of sarcoidosis at first presentation. This resulted in a considerable delay to diagnosis and to the corticosteroid therapy. CONCLUSION: The authors emphasize the rarity of the pseudoalveolar form of sarcoidosis. They insist on its roentgenographic characteristics and demonstrate the functional benefits allowed by the precocious medical management. They also propose a current review of the literature. PMID- 17127916 TI - [An acute interstitial pneumonia]. PMID- 17127917 TI - ["EXACO": four year follow-up of exacerbations in a cohort of patients with COPD]. AB - EXACO is a French prospective observational epidemiological study. Chest physicians will enrol 1200 patients who will be followed up for 4 years. The aims of the study are to describe the exacerbations in COPD patients at different levels of severity and to confirm the existence of a sub-group of patients defined as frequent exacerbators. The clinical characteristics associated with this profile and the decline of FEV1 will be evaluated. A validation of the VSRQ scale will be performed. The data will be collected from chest physicians and by phone calls to the patients every three months. PMID- 17127918 TI - [Yellow nail syndrome and transudative pleural effusion]. PMID- 17127952 TI - [Mediteranean environment and thoracic pathology]. PMID- 17127953 TI - [Evidence based medicine. The facts in the North countries]. PMID- 17127954 TI - [Evidence based medicine. The facts in the South countries]. PMID- 17127955 TI - [How to take into account the cost effectiveness parameters in the management of bronchial carcinoma?]. PMID- 17127956 TI - [Surgeon plays probably a pivotal role in the management of bronchial carcinoma]. PMID- 17127957 TI - [Is it possible to treat bronchial carcinoma without chemotherapy?]. PMID- 17127958 TI - [Listening to the patient]. PMID- 17127959 TI - [Evaluation of clinical practices in asthma]. PMID- 17127960 TI - [Evaluation and follow-up of asthma: the point of view of paediatrician]. PMID- 17127961 TI - [Education, is it the answer?]. PMID- 17127962 TI - [Why should we measure exhaled NO in asthma patients?]. PMID- 17127963 TI - [Biotherapies for the treatment of asthma: are they the treatment of the future?]. PMID- 17127964 TI - [Management of COPD: feasability]. PMID- 17127965 TI - [Organization and results of an information campaign and early diagnostic in Lebanon]. PMID- 17127966 TI - [Rehabilitation for COPD: programs and difficulties]. PMID- 17127967 TI - [COPD treatment by drugs]. PMID- 17127968 TI - [Anaphylaxis to anaesthesic drugs]. PMID- 17127969 TI - [Allergy to non steroidal anti-inflammatory drugs]. PMID- 17127970 TI - [Allergy to antibiotics]. PMID- 17127971 TI - [GARD (Global Alliance against chronic Respiratory Diseases)]. PMID- 17127972 TI - [How to make easier the treatment of asthma]. PMID- 17127973 TI - [Thoracic hydatid cysts: news]. PMID- 17127974 TI - [How long the antibiotic treatment should be for pulmonary infections?]. PMID- 17127975 TI - [New test for the diagnosis of tuberculosis]. PMID- 17127976 TI - [What is the place of surgery in the treatment of tuberculosis?]. PMID- 17127977 TI - [Is it possible to diagnose interstitial pneumonia only with lung imaging?]. PMID- 17127978 TI - [Idiopathic interstitial pneumonia: treatment]. PMID- 17127979 TI - [Surgery of the trachea: old problem, new possibilities]. PMID- 17127980 TI - [Malignant mesothelioma of the pleura: place of surgery]. PMID- 17128083 TI - [Proceedings of the Consensus Conference on Amyotrophic Lateral Sclerosis, 23-24 November 2005, Nice, France]. PMID- 17128084 TI - [What usual and unusual clinical signs suggest amyotrophic lateral sclerosis?]. AB - Diagnosis of amyotrophic lateral sclerosis (ALS) is typically restricted to clinical and electromyographical upper motoneuron (UMN) and lower motoneuron (LMN) features. However, the initial presentation is sometimes misleading. These unusual presentations are discussed. They can delay the diagnosis of ALS. PMID- 17128085 TI - [Classical and atypical clinical features in amyotrophic lateral sclerosis]. AB - Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder involving primarily motor neurons in the cerebral cortex, brainstem and spinal cord. In the absence of any biological marker, the diagnosis of ALS is based on clinical analysis, combined with the results of electromyography. Consensus diagnosis criteria (El Escorial criteria) have been developed to define workable and internationally acceptable guidelines for the diagnosis of ALS. A combination of lower and upper motor neuron signs with evidence of spread within a region or to other regions is required. The diagnosis of ALS has been categorized into various levels of certainty depending on the presence of upper motor neuron and lower motor neuron signs together in the same topographical anatomic region (brainstem and cervical, thoracic or lumbosacral spinal cord). Clinical types and patterns of ALS have been defined. The emerging concept of "ALS plus" is characterized by the presence of atypical clinical features, e.g. extrapyramidal signs or dementia, in association with the classical phenotype of ALS. This paper reviews the classical and atypical clinical features of ALS. PMID- 17128086 TI - [What are the clinical criteria of amyotrophic lateral sclerosis by clinical form?]. AB - The diagnosis of ALS requires the presence of lower and upper motor neuron degeneration and a progressive spread of symptoms and signs. Due to the lack of any biological diagnostic marker, the World Federation of Neurology established clinical and paraclinical criteria (El Escorial 1991; Airlie House 1998). These criteria allow to establish a diagnosis with various levels of certainty. In practice they have a low sensitivity, especially at the beginning of the disease, and may delay the diagnosis. They exclude also some clinical forms and are not helpful in the differential diagnosis. PMID- 17128087 TI - [Clinical characteristics of amyotrophic lateral sclerosis subsets]. AB - Amyotrophic lateral sclerosis (ALS) is characterized by significant clinical variability. Different subsets are classically individualized: bulbar onset and limb onset ALS, sporadic and familial ALS, ALS-plus syndromes (characterized by the presence of atypical clinical features, e.g. extrapyramidal signs or dementia, in association with the classical phenotype of ALS) and Western Pacific ALS. In addition, ALS-related syndromes include progressive muscular atrophy, primary lateral sclerosis and progressive bulbar palsy. The recognition of ALS subsets and ALS-related syndromes is important in clinical practice since the prognosis may vary depending on the clinical presentation. The prognosis of bulbar-onset ALS is poor compared with the spinal-onset type. Primary lateral sclerosis, defined by pure upper motor neuron findings, has a more benign course than classical ALS. It has also important implications for therapeutic trials to ensure the homogeneity of clinical material since inclusion of atypical forms with different prognoses can skew the outcome analysis. This paper reviews the clinical characteristics of the ALS subsets and ALS-related syndromes. PMID- 17128088 TI - [Electroneuromyographic criteria of amyotrophic lateral sclerosis]. AB - Electrophysiological study plays a key role in the diagnosis of amyotrophic lateral sclerosis (ALS). ENMG demonstrates lower motor neuron involvement in muscles clinically involved but also in non clinically involved territories. Conventional EMG shows mixed signs of active denervation, partial chronic denervation and fasciculation, in a variable pattern from one muscle to another. The choice of the muscles to be recorded will privilege muscles innervated by motor neurons from bulbar region, cervical, thoracic and lumbar medulla in order to define the extension of the pathological process. Special ENMG techniques (single fiber EMG, macro-EMG, motor unit firing study, motor unit number estimate) have been proposed for the evaluation and better understanding of pathophysiology. They are not used in routine. Motor conduction study demonstrates motor axons loss and excludes nerve conduction abnormalities of a demyelinating motor neuropathy. Sensory conduction and sensory nerve action potential amplitude are normal in ALS, even in severely motor involved territory. Electrophysiological study can show upper motor neuron involvement when studying motor evoked potentials using transcranial magnetic stimulation of motor cortex. Several parameters have been studied. They are diversely affected and so their diagnostic sensitivity also varies. PMID- 17128089 TI - [What electroneuromyographic criteria are required for the diagnosis of amyotrophic lateral sclerosis?]. AB - Electroneuromyography is essential for the diagnostic of amyotrophic lateral sclerosis. The aim of conventional needle examen is to detect peripheral motoneuron involvement by the identification of active and chronic diffuse neurogenic activities. This brings up the problem of the selection of the muscle to test to enhance the sensitivity of the exam. Recent techniques permit to study the reorganization of the motor units more precisely. Motoneuron loss is appraised by several methods for macro-EMG estimates of the number of motor units. The collateral reinnervation can be evaluated by single fibre EMG. The other main objective is to detect conduction abnormalities. These lead to differential diagnoses of pure motor neuropathy with conduction blocks. The different prognostic value of these different conditions required defining both sensitive and specific electrophysiological diagnostic criteria. PMID- 17128090 TI - [What is the role of other complementary examination in amyotrophic lateral sclerosis?]. AB - Amotrophic lateral sclerosis diagnosis is based on clinical and electrophysiological findings. Transcranial magnetic stimulation and MRI can show abnormalities which are not specific, but which can confirm upper motor neuron involvement. The other tests are performed to exclude differential diagnosis. Tests which should be performed in every cases are: medullar MRI, blood counts, erythrocyte sedimentation, serum protein electrophoresis, calcium, phosphore, serological tests for HIV, siphylis, Lyme disease. Other tests are made in some clinical circonstances to exclude genetical disease or metabolic disorders (SMN gene, Kennedy gene, Hexosaminidase A, very long chaine fatty acids), haematological or paraneoplasic disorders (anti-neurons antibodies, PSA, CT of chest and abdomen, mammography, bone marrow biopsy) or inclusion myositis (muscle biopsy). PMID- 17128091 TI - [Ancillary exams in the diagnosis of amyotrophic lateral sclerosis]. AB - The diagnosis of ALS is established on the basis of the revised El Escorial criteria revealing involvement of the upper and lower motor neuron. The presence of upper motor neuron signs is recognized by physical examination, but is not always easy to demonstrate. For the patient, early diagnosis reduces the uncertainty and the long waiting period before exclusion test can be performed. Early referral to ALS specialty clinics will have a beneficial effect on the patient's quality of life. Early diagnosis of ALS allows early use of drugs, slowing the rate of disease progression. MR proton spectroscopy and diffusion tensor imaging are potentially useful diagnosis tools but require further analysis of reproducibility in studies with more carefully matched patients and standardized techniques. There is no biochemical marker found in serum or cerebrospinal fluid to establish the definite diagnosis of ALS. Detection of Nogo A in muscle offers an easy tool for detecting the presence of ALS but further studies are needed to determine the specificity, sensitivity and predictive value of such modifications. When applying standard transcranial magnetic stimulation, examination of several territories improves sensitivity and the probability of detection of infraclinical upper motor neuron dysfunction. The cortical silent period seems most sensitive. The triple stimulation technique (TST) is a very sensitive method but needs to be confirmed by other teams. Some teams have stated that the technique is painful and time consuming for the patient. To date, the corticobulbar tract cannot be studied with TST and we have little experience for the lower limb in very few ALS patients. PMID- 17128092 TI - [Amyotrophic lateral sclerosis: differential diagnosis and frontier forms]. AB - Amyotrophic lateral sclerosis (ALS) may be mimicked by disorders affecting the different levels of the motor system from cortex to muscle. Clinical heterogeneity is a feature of both ALS and related syndromes allowing for a large differential diagnosis. During the initial stage of a motor disorder false positive and false negative diagnoses of ALS are possible. Examples of disorders that should not be misdiagnosed as ALS, because their prognosis and treatment differ, are multifocal motor neuropathy, Kennedy's bulbospinal atrophy, cervical myelopathy, hyperthyroidism and hyperparathyroidism. Syndromes remote from polio and radiation treatments should be recognised. Eventually, frontier forms of ALS with signs restricted to either the upper or lower motor neurons deserve particular attention. Electrodiagnosis is pivotal to disclose signs and extension of the peripheral motor neuron, to detect and quantify cortico-spinal involvement, to search for specific signs of conditions that mimic ALS. Until specific markers become available, clinical evaluation supported by electrodiagnosis and other ancillary tests are crucial to provide with the correct diagnosis, prognosis and treatment. PMID- 17128093 TI - [Differential diagnosis and atypical subsets of amyotrophic lateral sclerosis]. AB - Amyotrophic lateral sclerosis (ALS) is a progressive degeneration of upper and lower motor neurons. In the absence of any validated biological marker, the diagnosis of ALS depends upon recognition of characteristic symptoms and signs together with supportive electrophysiological findings. The diagnosis of ALS is easy to recognize in its fully developed form but during the early stages both false positive and false negative diagnoses are common. In clinical practice, diagnostic difficulties mostly arise with patients who present either with only upper motor neuron, or with only lower motor neuron signs. It may be difficult to distinguish ALS with clinically predominant lower motor neuron involvement from alternative diagnoses including spinal atrophies of adult onset, Kennedy's disease, inclusion body myositis and motor neuropathies with conduction blocks. The diagnosis of ALS related syndromes (progressive muscular atrophy, primary lateral sclerosis and progressive bulbar palsy) requires the elimination of alternate diagnoses. This paper reviews the main characteristics of diseases mimicking ALS and the atypical subsets of ALS. PMID- 17128094 TI - [What is the role of the genetic survey in amyotrophic lateral sclerosis?]. AB - The genetic inquiry in the diagnostic process of amyotrophic lateral sclerosis (ALS) need to be precise and systematic. Several hereditary neurologic disorders may initially mimic ALS, such as dominant spino-cerebellar ataxia or spastic paraplegia. Other hereditary motor neuron disorders are clinically more difficult to distinct from ALS such as Kennedy's disease, adult-onset spinal muscular atrophies or juvenile ALS. When a final diagnostic of ALS is established, the genetic inquiry aims at identifying a familial ALS (FALS) case. Almost 10 to 20 percent of ALS cases are familial. It is now admitted that 10 to 20 percent of those cases are due to SOD1 gene mutations. Such mutations may have a dominant or recessive inheritance. They are mainly found in multigenerational families. In 80 percent of the FALS cases, only two ALS cases are found in the pedigree. One cannot know whether this represent dominant cases with low penetrance, recessive inheritance or a multigenic, hereditary complex, disorder. More other, not all the SOD1 mutations have been demonstrated as causal. These elements are strong enough to suggest to precisely study, in the presence of a given SOD1 mutation, both the clinical phenotype, the data from the literature and, as often as possible, the segregation of the mutation into the family, before ascertaining that the mutation is responsible for a hereditary case of ALS. In every case, the genetic inquiry together with its conclusions should be done with caution, taking into account both the patient's need of a clear information and his anxiety regarding his descent. PMID- 17128095 TI - [Where is the role of the genetic investigations in amyotrophic lateral sclerosis?]. AB - About 10 p. cent of amyotrophic lateral sclerosis (ALS) cases are familial. Most of the familial ALS (FALS) cases are clinically homogeneous. Among these families, autosomal dominant, X-linked or autosomal recessive transmission can be observed. Most of the causal mutations have been observed in the SOD1 gene. To date, more than one hundred different mutations have been described, but it remains unclear whether the mutation is always responsible for the phenotype. Penetrance of the mutation depends on age, with almost 90 p. 100 of penetrance at age 70 years. There is no anticipation. Worldwide, the most frequent mutation is A4V with dominant transmission, responsible for a severe, rapid form of the disease. The second most frequent mutation is D90A which is generally transmitted recessively, predominantly in the Scandinavian countries. The phenotype is characterized by a long lasting course (mean: 11 years). Other causal mutations have been described in the Alsine, Apex, NF-H and NAIP genes. Other genes can be considered as risk factors, like SMN2, APO E4, APEX, Dynactine, P-450 D6. Presymptomatic testing for FALS seems difficult because little information can be given to the patient regarding the responsibility of the mutation in the disease, age of onset, and disease trends. The same precautions as for Huntington's disease are needed. Genetic investigations can contribute to better understanding of the pathophysiology of ALS. Other causal genes in the 90 p. 100 of FALS without SOD1 mutation and eventually in the sporadic ALS cases may be disclosed. Genetic investigations also determine the precise role of a given SOD1 mutation because of the large number of potential SOD1 mutations, the variability of the transmission mode and the non-exceptional absence of proven causality for ALS. PMID- 17128096 TI - [Psychological reactions to the announcement of a severe disease diagnosis: the amyotrophic lateral sclerosis example]. AB - To face the traumatic shock after the announcement of a severe and fatal disease, patients build psychological defences that physician must identify and respect as long as they don't endanger the patient's psychological fate and his medical care. ALS patient's do not have any curative perspective thus they will try "to escape" using different strategies the traumatic reality, that they cannot integrate and control. The physician has to prove his understanding and empathy facing these psychological reactions that could become of pathological appearance. The physician has to progressively bring the patient to consult the clinical psychologist; however he has not to forget that he is equally responsible about the way the patient go though his disease. PMID- 17128097 TI - [Announcement of amyotrophic lateral sclerosis diagnosis]. AB - Breaking the news of amyotrophic lateral sclerosis (ALS) is considered as a daunting task in most cases and is not a standardizable procedure. However, proven techniques exist to reduce the trauma to the patient. Announcing ALS falls upen the neurologist who must respect the ethical principle of the patient's independence. After the diagnosis is firmly established, the patient should be informed that he or she has a progressive disease of the motor nerves, for which no curative therapy is available. The name of the disease must be stated and explained. If the family history is negative, it is reassuring for the patient and family to know that their children are unlikely to be at risk. Positive aspects (no pain, no disturbances in sensation, cognition, memory and continence) should be stressed as well as the availability of efficient palliative measures for practically all symptoms. Current research efforts, and when available, the possibility of taking part in clinical studies of new drugs should be pointed out as a means of hope. The answer to the question of prognosis should include the information that there are no sudden worsenings to be expected, that the course of ALS may vary between months and decades, that making a firm statement on prognosis all but impossible for any single patient and that respiratory function may worsen during the disease course. It is therefore mandatory to inform patients and families about the existence of ALS patients'associations. The way the patient is told the diagnosis is of great importance and is considered as a multiple-step procedure. Discussion should take place in a private and quiet room and respect some fundamental objectives such as finding out what the patient already knows or suspects and how much more the patient wants to know, observing and responding to the patient's reactions, reinforcing the information and planning the future. It is proven that communicating the diagnosis of ALS in an empathetic fashion is an important and sensitive step to disease management. PMID- 17128098 TI - [How should the diagnosis of amyotrophic lateral sclerosis be announced?]. AB - In amyotrophic lateral sclerosis (ALS) announcement of the diagnosis is an important step in the process of comprehensive care. The patterns of psychological reactions following bad news must be considered with precaution. The neurologist must take into consideration the specific aspects of announcing familial ALS. This paper reviews the modalities of announcement of the diagnosis and course in ALS patients. PMID- 17128099 TI - [Methods of the announcement of amyotrophic lateral sclerosis diagnosis in familial forms]. AB - Between 10 percent and 20 percent of amyotrophic lateral sclerosis cases are familial (FALS). The announcement of a diagnosis of ALS to a patient having relatives already affected with this disease must take into account the familial history and should focus on the improvement in management of ALS patients. Diagnosis of FALS implicitly raises the notion of genetic factors and the possibility for an heritability of the disease (risk of ALS for relatives). Although it is possible to search for mutations in SOD1 gene in ALS patients and asymptomatic relatives, the incomplete penetrance of the disease, the low percentage (10 to 20 percent) of FALS linked to SOD1 mutation and the existence of non causal SOD1 mutations complicate the conclusions of genetic investigations concerning the real risk for a relative with a SOD1 mutation to develop ALS. PMID- 17128100 TI - [Breaking bad news during the course of amyotrophic lateral sclerosis: how and when]. AB - Operatory procedures can help to break bad news in ALS but the main goal of this unpleasant task stands in the quality of the relationship between the physician and his patient which must be preserved throughout the course of the disease. Plasticity is obviously the best way to characterize the appropriate kind of care in such a variable disease. The caregivers and the psychological factors must also be taken into account. As it is the case for therapeutic research, breaking bad news in ALS requieres a specific know-how which should be ideally performed by specialized teams within the framework of dedicated ALS centers. PMID- 17128101 TI - [Functional scales and motor assessment in amyotrophic lateral sclerosis]. AB - There is a need of standardized and quantitative clinical assessments of ALS patients receiving new treatments. Some clinical scales, that are based on patients interviews and/or on physical examination, allow to quantify the patient functional status over time. The ALSFRS has been validated for that purpose and is one the most commonly used scales in clinical trials. Its revised version (the ALSFRS-R) has recently shown excellent metric properties. Functional scales are part of the clinical evaluation of ALS patients, and complete the quantitative assessment of muscle strength, that includes the Manual Muscular Testing (MMT) and the Maximal Voluntary Isometric Contraction (MVIC). Readers will find in this article a (non-exhaustive) listing of functional and motor scales that can be useful both in clinical trials of experimental agents and in daily practice. PMID- 17128102 TI - [Evaluation of strength in amyotrophic lateral sclerosis]. AB - The hallmark of progression in patients with amyotrophic lateral sclerosis (ALS) is the development of progressive weakness and muscular wasting. Strength testing has been used to monitor the course of the disease and to test the efficacy of new drugs. This paper review the methods used to quantify the weakness and compare their accuracy and reproducibility. PMID- 17128103 TI - [Amyotrophic lateral sclerosis: cognitive and behavioral evaluation]. AB - Cognitive impairment in nondemented ALS patients has been demonstrated, although its incidence remains to be determined. FTD is the most frequently form of dementia in ALS. The clinical profile of patients with dementia or mild cognitive deficit evokes neuropsychological deficits and behavioural changes resulting from executive dysfunction. The psychometric evaluation, centred on executive disturbances, goes with behavioural scales in order to accurately appreciate the repercussion of cognitive and behavioural changes on daily life. PMID- 17128104 TI - [Neuropsychological assessment in amyotrophic lateral sclerosis]. AB - Neuropsychological assessment in the ALS population is relatively recent probably because this disease was traditionally considered to spare cognitive abilities. However, evidence is emerging from neuropsychological testing that some patients with classical ALS, who are thought not to be demented, develop cognitive deficits and particularly frontal executive disturbances. Neuroimaging gives support to frontal lobe involvement in ALS. Nevertheless, cognitive impairment does not concern all ALS patients and appears to be mild when it is described compared with rather severe frontotemporal dementia observed in association with ALS. Most of the references in this work question the cognitive perspective in the ALS population but few of them can provide a real useful tool for cognitive investigation in this disease. PMID- 17128105 TI - [Psychological and psychopathological evaluation in amyotrophic lateral sclerosis]. AB - In the literature, few studies refer specifically to the psychological and psychopathological aspects of the ALS. However most of these studies mention the psychological specificities observed in ALS patients. In fact, they are not depressed, nor anxious, and they seem to accept surprisingly well the threathening diagnosis. Some authors have mentionned the presence of denial as an adaptive mechanism. It remains very difficult to compare the various studies. First, the stage of the evolution of the disease and the physical abilities of the patients differ greatly; in fact it has been demonstrated that at each stages the patient present specific psychological concerns. Second, the psychological concepts refer to different level of understanding. Some authors study depression or anxiety, others focus on the global psychological status and others focus on a psychological mechanism, such as denial for example. Third, the scales used to assess these psychological or psychopathological aspects are, on one hand, never specific to ALS and, on the other hand, are sometimes filled by the clinicien or sometimes self reports filled by the patient, his caregiver or even by the medical staff. Thus, many studies have created their own scales or interview, which are not validated. So, it appears necessary, in order to be able to compare the different results, to use validated scales which are recognized by the scientific community, while waiting for specific tools created and validated in populations of ALS patients. PMID- 17128106 TI - [Psychological and psychopathological assessment of the patient with amyotrophic lateral sclerosis]. AB - Management of patients with amyotrophic lateral sclerosis (ALS) implies the participation of many health care professionals. The needs and goals of the patients cannot be limited to functional factors and should be considered in their full psychological, social and familial settings. Thus, much of the care for patients with ALS must focus on symptom relief, comfort and psychological support in order to improve their quality of life (QOL). Studies have demonstrated the significant role of psychological, existential and supportive factors in determining QOL in these patients. This work deals with psychological assessment in patients with ALS and reported studies within this population. Psychological assessment may vary and its methodology depends on the goals. Most frequently, psychological assessment implies depression and anxiety, coping strategies or quality of life. PMID- 17128107 TI - [Nutritional assessment in amyotrophic lateral sclerosis patients]. AB - During ALS, malnutrition may occur. The causes are multiple. A reduction in caloric and protein intakes, due to swallowing disorders, play a major role. Moreover, (2/3) of patients develop an hypermetabolism of unknown cause, leading to an increase in resting energy expenditure. Malnutrition, whose prevalence is 10-55 percent, is an independant significant prognostic factor for survival. Nutritional assessment is recommanded every 3-6 months at least, and more often if needed. After a simple clinical examination, nutritional assessment includes the measurement of weight (W) and height (H), in order to calculate the Body Mass Index (BMI=W/H(2)). BMI<18.5 (age: 18-65ys) or<20kg/m2 (age>65ys) indicates malnutrition. A weight loss more than 5 to 10 percent of usual weight in the last six months is another malnutrition criterion. The measurement of skinfolds and the calculation of mid arm muscle circumference give informations on respectively fat mass and fat-free mass, but their interest is mainly during the follow-up, or for triceps skinfold when included in a validated bioimpedance formula. Bioimpedance analysis (BIA) is a simple bedside technique, recently validated for measurement of fat-free mass in ALS patients. The BIA phase angle could be an easy obtained parameter of severity. Dietary interview is important when patients still use oral route, but is difficult if they have elocution or writing problems. Indirect calorimetry and dual X-ray absorptiometry give reliable informations on respectively energy expenditure and body compartments, but remain scarcely used. Additional methods, like swallowing tests, psychological, digestive or respiratory assessments are often useful for optimal nutritional prescriptions. We recommand as minimal nutritional assessment a systematic clinical examination, anthropometric measurements and bioimpedance evaluation. PMID- 17128108 TI - [Nutritional approach for patients with amyotrophic lateral sclerosis]. AB - Prevalence of malnutrition in ALS ranges between 16 and 53 p. 100. The percentage of loss of weight greater than 10 p. 100 or, in a more inconstant way, body mass index lower than 18.5, are relevant criteria of malnutrition because predictive of survival. Arm muscle circumference and bioelectrical impedance analysis can assess body composition, but their impact on disease progression has not been evaluated. Measurement of serum albumin levels is of no interest. NUTRITIONAL REQUIREMENTS: Considering that energy expenditure is increased 10 to 20 p. cent in the majority of patients, energy needs can be estimated to be approximately 35 kcal/kg/d. In order to limit the accumulation of fat mass in the sub-group of patients with decreased energy expenditure, monitoring of triceps skin fold or impedance could be proposed, but their interest remains to be evaluated. In the absence of available data on protein requirements in ALS, an intake ranging between 1 and 1,5 g/kg/d seems reasonable, knowing that there is a risk of deficiency when intake is less than 1 g/kg/d and that an increase to 1,5 g/kg/d, considered as harmless, could be useful in the event of hypercatabolism. Supplementation with creatine, antioxidants or amino acids has not proven to be effective. These nutrients being provided by a balanced diet, specific supplementations have no proven utility provided that the patient receives sufficient proteins and energy. Systematic supplementation with vitamin D is however warranted to prevent osteoporosis due to the known risk related to a common state of deficiency. RELEVANCE OF GASTROSTOMY: Studies with good methodological quality evaluating percutaneous endoscopic gastrostomy (PEG) in ALS are not available. Because of the impact of malnutrition on survival, PEG must be considered when oral intake becomes insufficient. Retrospective studies suggest that the PEG tube is usually inserted too late during the disease course, minimizing expected benefits. Criteria useful for making the decision to installation a PEG tube should be the subject of randomized controlled studies. Mortality in the month which follows PEG, approximately 10 p. cent, is primarily due to respiratory failure. It occurs more frequently among patients having a forced vital capacity<50 p. cent. To avoid a degradation of pulmonary function in this sub-group of patients, several approaches are suggested in the literature: earlier PEG, peri-operative noninvasive ventilation, and radiological gastrostomy which does not require a general anesthesia. Whatever the technique used for gastrostomy, it should be carried out in an expert centre, because patients with ALS are more exposed to complications than others. PMID- 17128109 TI - [Amyotrophic lateral sclerosis (ALS): evaluation of respiratory function]. AB - Respiratory involvement is an almost constant feature of als, with a usually rapid progression leading to respiratory failure. These characteristics justify a close follow up, usually at three-month intervals. A systematic, careful clinical evaluation is essential to detect the subtle respiratory symptoms and signs related to respiratory muscle failure. Dyspnea and orthopnea are often late findings in patients with a usually severe functional impairment due to peripheral muscle weakness. Nocturnal respiratory events (obstructive sleep apnea syndrome and hypoventilation) are strongly suggested by daytime hypersomnolence and frequent morning headaches. Physical evaluation is essential to detect accessory muscle recruitment, supine abdominal paradox, and encumbrance of upper or lower airways. Vital capacity (VC) is the most classical lung function test. The major limitation of spirometry is its poor sensitivity to detect a moderate inspiratory muscle weakness. Supine VC may improve the detection of diaphragmatic involvement. Peak expiratory flow during cough (cough PEF) gives an overall evaluation of cough efficiency, values below 160 to 270 L/min suggesting poor airway clearance. Arterial blood gases are performed at first evaluation and subsequently in case of clinical signs, significant deterioration of lung function tests, or sleep desaturations. Hypercapnia is weakly related to lung function results in bulbar patients. A specific evaluation of respiratory muscle strength is mandatory, as these tests are both sensitive and highly prognostic. Possible discrepancies (particularly in bulbar patients) between Maximal inspiratory pressure (PImax) and sniff nasal inspiratory pressure (SNIP) justify to perform both measurements and to select the highest pressure. A maximal expiratory pressure (PEmax) below 45 cm H2O may indicate a compromised cough efficiency but the correlation with cough PEF may be poor. Screening nocturnal oxymetry is useful to detect sleep apneas and hypoventilation. Criteria defining significant desaturations remain however controversial. Suspicion of obstructive sleep apnea syndrome on clinical grounds or oxymetry findings should be confirmed by a conventional polysomnography. PMID- 17128110 TI - [Evaluation of ventilatory functions in amyotrophic lateral sclerosis]. AB - As the clinical signs of amyotrophic of lateral sclerosis (ALS) are late and not specific, spirometric measurements, study of respiratory strength muscles and analysis of night respiratory parameters are used to evaluate ventilatory functions. The prognostic value of vital capacity is well established, but with a poor sensitivity to detect the initial form of ALS. Gazometric parameters remain normal for a long time. Hypercapnia is late and due to a major reduction in the force of the inspiratory muscles. Level of diurnal venous bicarbonate could detect nighttime hypoventilation. Noninvasive techniques (Pimax, Pemax, snifftest) are available to explore the strength of respiratory muscles. Sleep is characterized by a long period of hypoventilation, essentially during REM-sleep. There are no guidelines concerning the appropriate mode, type and rhythm of explorations of diurnal and nocturnal respiratory functions. PMID- 17128111 TI - [Handicap and quality of life assessment in amyotrophic lateral sclerosis]. AB - Measures of handicap progression alone are insufficient for assessing what a patient experiences. Handicap results from the interaction between a person's disabilities and the social, familial and environmental factors which determine his or her living conditions. Other factors condition quality of life: of these, psychological and existential factors and an individual's ability to adapt play a fundamental role. Measuring handicap and quality of life must provide medical care providers with comprehensive knowledge of the way in which an individual experiences the disease, his or her handicap and the side effects and constraints of treatments within his or her own life context. PMID- 17128112 TI - [Handicap and quality of life evaluation in amyotrophic lateral sclerosis]. AB - The impact of amyotrophic lateral sclerosis and consequent disability on everyday life can be assessed with generic or specific and/or functional quality of life scales. Generic scales SF36, SIP (Sickness Impact Profiles) and SIP/ALS 19, SEIQoL have been validated for the assessment of quality of life in SLA. A specific scale has also been worked out and validated in this disease, the ALSAQ 40 scale. The ALSFRS (ALS Functional Rating Scale) is a tool validated to evaluate the patient's functional capacities. However, the patient's quality of life felt does not depend only on the disease course but also the patient's previous psychological profile, the environmental, social conditions and spiritual aspirations. The absence of a cognitive deficit makes the patient particularly lucid about the progressive degradation of health status. Evaluation must be carried out with individually for each patient, taking into account the social and familial environments, to appreciate real living conditions, and the impact of this degenerative disease. PMID- 17128113 TI - [Caring for a patient with amyotrophic lateral sclerosis: the patient's and the caregiver's experience]. AB - ALS usually affects highly active, working, sportive individuals. The disease has a devastating impact on the patient's personal life and family ties. Indeed, the disease requires constant attention from the carer for its rapidly progressive nature leads the patient to a complete state of isolation without affecting his/her intellect. For the patient days, are numbered, for the carer, time is limited. The carer must cope with a number of medical, administrative and financial difficulties. These include belated diagnosis with its disastrous consequences on medication, lack of response from the decision-making services, home-improvements or car-fitting. All this requires amazing tenacity from the carer. All the more so as the progress of handicap is relentless and the patient is bound to use different forms of support, walking-stick, zimmer, wheel-chair, hoist, speech synthesis, gastrostomy, tracheotomy or palliative care, which turn out to be more or less effective. But mechanical devices are not enough. Assistance by a nurse, a physiotherapist, a speech therapist and/or a nursing auxiliary is necessary. There are not enough of these assistants and the result cannot be guaranteed, but their role is essential in home care. The carer's investment can be terribly consuming. The burden of 24h care wears out the carer whose days and nights are being regulated by the disease. Not to mention lonely patients with no means of support who find themselves confronted with inevitable daunting problems Institutions providing either temporary or permanent care for ALS patients are scarce. As the condition worsens steadily, the patient suffers sheer desperation and guilt feelings and resents being a burden for family yet would rather stay with them. ARS volunteers who collaborate with medical teams can provide much help. The carer's burden should be greatly lightened too by the newly-installed tutorial system: patients' files will be followed up by one single person after the center's social worker has drawn them up. Medical and home volunteers cooperate to give the patient the feeling that far from being abandoned or ostracized by society, he/she is constantly being given care. PMID- 17128114 TI - [What are the etiological medical therapies in amyotrophic lateral sclerosis?]. AB - The increasing knowledge about Amyotrophic Lateral sclerosis (ALS) led to the development of the first and only available treatment: riluzole. The efficacy of this drug has been demonstrated in two controlled clinical trials. The data showed that riluzole increases the rates of survival in patients suffering from ALS. This increase was about 25 percent over a period of 12 months of treatment. Since its marketing, the clinical efficacy of riluzole was confirmed in several epidemiological studies. Tested initially because of its anti-glutamates qualities, the exact mechanisms of action responsible for the efficacy of Riluzole in ALS remains unknown. Furthermore, despite a strong rational based on important literature, several other anti-glutamates drugs failed to demonstrate efficacy in ALS, nor did numerous anti-oxydant, neurotrophics, immunomodulators, and favoring mitochondrial metabolism drugs. Despite the low rate of patients showing an increase of liver enzymes, a strict control of liver functions is necessary with a treatment by riluzole. its dose should be increased gradually in case of important fatigue, a change of the general state, or respiratory incapacity. Because of the neuroprotectrives qualities of riluzole, early neuronal death in ALS, the difficulty to confirm early the diagnosis, and the evident interest to treat in the first stage, the treatment of the suspected forms of the disease becomes justifiable. The perspectives of treatment of ALS will depend on our capacity to understand better the physiopathology, the reasons of failures in the past, and to develop new biomarkers. They will depend also on our capacity to include in clinical trials and to follow up over a long time, an important number of patients. Different new methods, as the cellular therapy are in development, and represent a real hope for the future. PMID- 17128115 TI - [Etiological drug therapy for amyotrophic lateral sclerosis]. AB - Progress in the last decades has greatly strengthened our understanding of the etiopathogenic mechanisms of amyotrophic lateral sclerosis. Such progress has lead to an evaluation of several compounds affecting excitotoxicity, oxidative stress, apoptosis, growth factors and inflammation. Riluzole has clearly proven efficiency on mortality with a 35 p.cent reduction of death or tracheotomy at 18 months. While the mechanism of action of this compound remains to be fully elucidated, there is evidence in favor of a neuroprotective effect. Other compounds have been tried in ALS. To date, results have been disappointing with worsening even being noted in some cases. These negative studies offer some insight into the pathophysiological mechanism regulating the disease, raising pertinent questions concerning the transposition of animals models to humans and about interractions between these different compounds and riluzole. PMID- 17128116 TI - [Symptomatic treatments in amyotrophic lateral sclerosis]. AB - ALS is a progressive, fatal, degenerative motor neuron disease of unknown cause. Although advances in understanding pathophysiology of ALS have stimulated the development of new therapies, most of them remain few efficient or ineffective and the main management of ALS patient, to improve quality of life by alleviating symptoms, is symptomatic treatment. This article discusses the approaches now in use to manage some of the most common symptoms of ALS including the following: spasticity, cramps, pain, laryngospasm, pseudobulbar syndrome, salivation and drooling, sleep disorders and fatigue, constipation and trophic troubles. PMID- 17128117 TI - [Drug therapy for symptomatic relief in amyotrophic lateral sclerosis]. AB - Amyotrophic lateral sclerosis is a neurodegenerative disease that has no curative treatment. However, some symptoms of the disease can respond to specific treatments. The aim of these treatments is to enhance the patient's quality of life and in some instances survival. Besides physical therapy, and nutritional and respiratory supportive systems, several specific medications can be useful. However, despite the frequency of these symptoms, few studies have evaluated the benefit of these medications in ALS. Their use is most often based on clinical experience or on studies conducted in other neurological diseases. So, for most of these medications, available evidence does not permit a precise evaluation if their impact on quality of life and survival. PMID- 17128118 TI - [What physical therapy techniques can be used to improve airway freedom in amyotrophic lateral sclerosis?]. AB - In individuals with ALS rehabilitation is mainly designed to prevent fatigue and contracture, to improve independence and activities for as long as possible, to optimize ability to live with the handicap, and finally to maximize quality of life. The functional impairment must be defined and physical therapy techniques have to be adapted to each patient and reevaluated frequently during the course of the disease. Various types of massage and exercise, monitored by a physical therapist are effective. Strengthening or endurance exercises are controversial as exercise may injure muscle fibres and motor neurons. Isometric exercise, short of fatigue, of unaffected muscles is recommended. Range of motion exercise is critically important for preventing contraction. Assistive and adaptative equipments are essential for maintaining the patient's activities of daily living and home equipment preserves independence. Several orthoses for hand, arm, foot or cervical weakness are available. A wheelchair is an important adaptative device when walking becomes too fatiguing or impossible. Choice for special options and features may require attention. Pulmonary complications are prevented with adapted techniques for bronchic obstruction. Based on the degree of weakness of limb and axial muscles six stages of functional impairment can be defined ranging from fully ambulatory in stage I to bedridden and totally dependent in stage VI. This staging provides a framework for physical therapy evaluation and guidance for appropriate rehabilitation in ALS patients. PMID- 17128119 TI - [Physical therapy in amyotrophic lateral sclerosis]. AB - Amyotrophic lateral sclerosis is a neurodegenerative disease, without any curative treatment. Clinical expression is variable and related to loss of motor neurons in the cortex, brain stem and spinal cord. There is little scientific evidence demonstrating the usefulness of physical therapy in this disease. Only stretching exercises, proprioceptive neuromuscular facilitation techniques and functional mobility training seem to have a real benefit in terms of spasticity, quality-of-life and pain. The main objective of physical therapy appears to be the preservation of optimal quality-of-life throughout the course of this incurable degenerative disease. PMID- 17128120 TI - [Techniques favoring airway clearance in patients with amyotrophic lateral sclerosis]. AB - Although noninvasive ventilation may improve survival in patients with amyotrophic lateral sclerosis (ALS), ineffective airway clearance is an important cause of therapeutic failure. We report in this paper the main studies which have assessed assisted cough techniques in patients with ALS. Manually assisted cough (in particular with previous air stacking) and mechanical insufflation/exsufflation may significantly increase cough peak flow. Characteristics, limitations and long-term benefits of these techniques are also discussed. PMID- 17128121 TI - [Types of ventilatory support and their indications in amyotrophic lateral sclerosis]. AB - Respiratory muscle weakness represents the major cause of mortality in patients with amyotrophic lateral sclerosis (ALS). As a result, ventilatory assistance is an important part of disease management. Nowadays, noninvasive ventilation (NIV) has become the first choice modality for most patients and represents an alternative to tracheostomy intermittent positive-pressure ventilation. Although, some consensus guidelines have been proposed to initiate NIV in patients with restrictive chronic respiratory failure, these criteria are discussed regarding ALS. While the current consensus recommends that NIV may be used in symptomatic patients with hypercapnia or forced vital capacity<50p.cent of predicted value, early use of NIV is proposed in the literature and reported in this paper. PMID- 17128122 TI - [What is the role of life support systems in amyotrophic lateral sclerosis?]. AB - There is very little information in the medical literature concerning the problems of tracheotomy and emergency intubation in amyotrophic lateral sclerosis. Nevertheless, we can assumed that: explicative information for the patient is important, emergency ventilation is not very frequent, emergency ventilation was not systematic, prognosis of emergency ventilation is very poor. PMID- 17128123 TI - [Speech therapy guidelines in patients with amyotrophic lateral sclerosis]. AB - Bulbar involvement is usual in patients with amyotrophic lateral sclerosis (ALS), leading to a progressive loss of speech, phonation and swallowing abilities. The goal of the speech therapy is to maintain, as long as possible, those abilities and to devise alternative communication strategies when oral communication is ineffective. This paper presents guidelines for speech therapy during disease progression. PMID- 17128124 TI - [What are management practices for speech therapy in amyotrophic lateral sclerosis?]. AB - Amyotrophic lateral sclerosis involves deterioration of speech and swallowing. The objectives of rehabilitation are to maintain a comprehensible speech, using exercises on airflow-phonic coordination and breath, lingual and labial movements. Rehabilitation exercises also focus on the different times of swallowing to improve management of the food bolus in the mouth, to learn facilitating techniques and to adopt postures favoring passage of the food bolus. No studies have addressed the question of the effectiveness of rehabilitation in amyotrophic lateral sclerosis, in term of quality of life, maintenance of comprehensible speech or reduction of respiratory risks. Rehabilitation programs are established on an individual basis according to the clinical presentation and course. PMID- 17128125 TI - [Management of lost independence and its social consequences in amyotrophic lateral sclerosis]. AB - Amyotrophic lateral sclerosis is characterized by progressive unavoidable loss of independence but with preserved intellectual capacity. Care requires human and technical compensation taking into account the desires of the patient and his/her family. Care should be taken to avoid isolating from the social environment by implementing coherent material and human organizations aimed at maintaining patient comfort and acceptable quality of life "to the end" while respecting the rights and will of the care recipient. PMID- 17128126 TI - [The coverage of the social consequences of the disease and the loss of autonomy in amyotrophic lateral sclerosis]. AB - There is no specific social answer to amyotrophic lateral sclerosis, which alters quickly autonomy of the patients. Different financial and human assistances can be set up to compensate the loss of autonomy. To date, to obtain this assistance, delays are often long; and there is no individual evaluation of the situation at home. The creation of reference centres should allow a better coverage focused on an individualized answer with a management of systems based on the nearness. PMID- 17128127 TI - [Role of health volunteers for patients with amyotrophic lateral sclerosis and their family]. AB - In ALS Centers, the patient receives coordinated care from Health Volunteers and the medical team including specialized physicians, therapists and social workers. There are two types of volunteers: those who assist the patient in hospital, the Medical Volunteers (MV), and those who make home visits, the Home Volunteers (HV). Both are recruited and trained by the ARS and have a single motivation: help the patient's; they are emotionally and morally fully qualified to accomplish their task in accordance with the rules that have been set by the Association. The ALS patient is seen for the first time by the Medical Volunteer at the ALS Center, immediately after his or her disease has been diagnosed by the clinic's director. The MV takes the initiative for the dialogue, which he/she leads gently and tactfully. The discussion is aimed at gathering useful information to be put in the document: "Connaissance du malade" (knowledge of the patient), working out the patient's primary concerns and assessing the carer's commitment, in a friendly atmosphere. Finally the volunteer gives the guidebook, "Livret d'accueil" to the patient. After the conversation, the MV decides which confidential pieces of information are to be passed on to the medical staff with a view to sorting out problems; information is then set the HV who will provide follow up care for this patient. Long-term care management of ALS patients is achieved through three regular monthly appointments at the ALS Center. With the help of the document "Connaissance du malade" the MV keeps better in touch with the patient using the feedback from the HV about what has been going on in the interval between the two visits to the ALS Center. This collaboration enables appropriate follow up care for the patients with clearly-defined objectives: dealing with the patient's anxiety, understanding his/her position at the present time, his/her daily routine and needs, comforting, checking compliance with treatment, counselling and supporting the family, answering delicate questions. Thus, the Health Volunteer's mission contributes meaningfully to medical treatment. This approach likely helps the person affected by ALS to regain the initially deteriorated sense of belonging to the social body. PMID- 17128128 TI - [What is the role of health volunteers in amyotrophic lateral sclerosis?]. AB - OBJECTIVE: Analysis of the literature about functions of health volunteers in ALS. METHOD: Article selection in MEDLINE, CINHAL and EMBASE. RESULTS: The notion of health volunteer has several meanings, and relates to different models varying between substitution and supply. The role of volunteers in a French and two Canadian NPO are described. CONCLUSIONS: The function of the volunteer integrates potentially interpersonal aid relationships, social mediation, and active functional help components. PMID- 17128129 TI - [Psychological treatment for the patient and caregivers during the course of amyotrophic lateral sclerosis]. AB - Amyotrophic lateral sclerosis is a devastating disorder for which the psychological consequences of both the diagnosis announcement and the evolution of paralysis not only concern the patient but also his family. The role of the psychologist is to develop an individualized follow up considering the patient in his globality. The first consultation is, ideally, initiated after a medical consultation explaining the importance of the psychological area in ALS patient care. The psychological follow up will consist in an empathic listening of history and problems. Information will also be given to the patient by the psychologist who's role should not be only passive. When talking about "globality" of the psychological intervention for a given patient, his family takes a determinant place. The psychologist should be able to establish a contact with the family members concerned by the daily support to the patient. The psychological processes through which a patient will evolve should be explained to the family. Depression frequently affects family members, and a specific follow up in those cases has to be undertaken as soon as possible. Such a depressive reaction may also take place after death and a psychological follow up do not end after the death of a patient. The role of a psychologist in ALS care ideally takes places in the context of a multidisciplinary team such as a motoneuron clinic now largely available in our country. The burden of care is frequently heavy both for the family and the team of professional carers into and outside the hospital. The psychologist has a role of mediation between those persons, facilitating verbal exchanges, paying attention to specific difficulties and maintaining a fruitful exchange between the carers, the patient and his family. More prospectively, the psychologist also has a pedagogic role for the carers explaining psychological processes and giving clues for a constructive relationship between the patient and his family and also between this patient and his carers. PMID- 17128130 TI - [Psychological management of patient with amyotrophic lateral sclerosis and their caregivers]. AB - Psychological management of patients with amyotrophic lateral sclerosis (ALS) and their caregivers is a relatively recent concern, probably because there was previously a tendency to consider only physical functioning. Recent work have focused more on psychological well-being concerning patient and their close relative and have highlighted the great need for considering psychological reactions after such a diagnosis. The needs and goals of the patients cannot be limited to functional factors and should be considered in their psychological, social and familial settings. Currently, we consider that close relatives should benefit from psychological support just like patients with ALS. This review suggests different ways to improve management of patients with ALS, while considering personal for strategies coping with the disease, the familial and social context, and individual factors that could improve quality of life (QOL). Studies have demonstrated the significant role of psychological, existential and support factors in determining QOL in these patients. PMID- 17128131 TI - [What are the means of alimentary function supply and their indications in amyotrophic lateral sclerosis?]. AB - Artificial nutrition is necessary when oral feeding becomes insufficient to cover protein and energetic needs and becomes dangerous (risk of malnutrition, dehydration and aspiration). In ALS patients, enteral nutrition is the method of choice and gastrostomy is preferable to nasogastric tube which must be limited for a short term enteral nutrition or if gastrostomy is at risk (because of pulmonary function) or refused by the patient. The percutaneous gastrostomy can be placed endoscopically (PEG) or radiologically (RIG), surgical gastrostomy has to be avoided because of general anaesthesia. Advantages of RIG are a success rate of about 100 percent and a placement feasible without sedation but its superiority on PEG in ALS patients especially if pulmonary functions are altered is not demonstrated. No objective criterion permits to define the exact moment of enteral nutrition. However, enteral nutrition is recommended when dysphagia becomes symptomatic (insufficient caloric intake with weight loss, dehydration, frequent choking and aspiration). Swallowing disorders must be detected early to give to patients and their family information about enteral nutrition and gastrostomy as soon as possible and to help them to decide. It is desirable to propose gastrostomy when forced vital capacity is yet above 50 percent and nutritional state not altered (body mass index>18kg/m2 and/or weight loss<10 percent). Enteral nutrition is not desirable in ALS patients with dementia or in the preterminal phase. Suitable enteral nutrition with regular nutritional evaluation can improve nutritional status. Currently, improvement of quality of life and survival due to enteral nutrition has not been proved in ALS patients. PMID- 17128132 TI - [Indications and equipment needs for ventilatory support in amyotrophic lateral sclerosis]. AB - Long term mechanical ventilation improves survival and quality of life in patients with Amyotrophic Lateral Sclerosis (ALS). There is an enormous range of equipment available but there is little evidence pointing to the superiority of one device over another. Another problem is the lack of adaptation of equipment for use by handicapped patients. Indication on mechanical ventilation in ALS is discuss in this article: Symptoms associated with an objective criteria of hypoventilation are often used. In the monitoring of this mechanical ventilation new tools are needed, initiating nocturnal ventilation with systematic respiratory polygraphic studies seems to be justified. PMID- 17128133 TI - [Questions concerning an endotracheal intubation or a tracheotomy in amyotrophic lateral sclerosis]. AB - Amyotrophic lateral sclerosis (ALS) is an illness in which respiratory complications often determine the terminal prognosis. Emergency situations lead one to pose questions concerning an endotracheal intubation or a tracheotomy. A tracheotomy should not be performed during an emergency situation. A tracheotomy necessitates a stable condition and prior reflection. Orotrachael intubation is the method of choice during emergency situations requiring invasive ventilation or airway protection. Intubation during an emergency situation presents specific problems: the lack of knowledge concerning the person and their pre-established desires, the impossibility of evaluating the potential reversibility of an acute pathology, the risk of not being able to wean the patient off the ventilator and the lack of time to gather all the elements necessary for a well-thought out decision. It may be appropriate for emergency personnel to introduce mechanical ventilation and leave the reflection for a later moment, but this approach is not suitable for people in end of life situations in which the person and the family wish to avoid all unreasonable therapeutics. One solution may be to develop among emergency care teams the practice of using non-invasive ventilation and airway clearance techniques as well as developing palliative care knowledge. Orotracheal intubation in an emergency situation presents certain practical difficulties, notably regarding the choice of anesthetics. Preventings situations where emergency intubation may be necessary is probably best obtained by anticipating acute problems, by preparing the ill person, the family and the care givers, by coordinating the potential care providers and by educating emergency personnel in palliative care. PMID- 17128134 TI - [Ethical aspects for substitute decisions and their reversion in amyotrophic lateral sclerosis]. AB - Related to its clinical characteristics, amyotrophic lateral sclerosis (ALS) is a paradigm of the ethical difficulties due to a chronic and relentless fatal disease. Its fatal evolution in relation with a respiratory failure, but mainly the choices imposed by the respiratory insufficiency at the end of live, makes that the medical team faces very difficult situations. These situation can be solved only case per case, with repeated discussion with the patients and its relatives. The appeal to the shared dossier becomes essential in this approach. It makes easier the communication, and the appropriation and sharing of information and knowledge that are necessary to anticipate the end of life decisions. ALS sparks off constantly renewed dilemmas. PMID- 17128135 TI - [Ethical issues and supportive tools and their reversibility in amyotrophic lateral sclerosis]. PMID- 17128136 TI - [HAS recommendations on amyotrophic lateral sclerosis]. PMID- 17128137 TI - [HAS recommendations on amyotrophic lateral sclerosis]. PMID- 17128150 TI - [MUC1 and its applications in diagnostic pathology]. PMID- 17128151 TI - [Neuromuscular biopsy and diagnosis of vasculitis]. AB - One characteristic histological lesion on biopsy specimens is mandatory to establish the diagnosis of vasculitis. Combined nerve and muscle biopsies, by the same cutaneous incision, improve significantly the percentage of positive results. Nerve fragments should be taken in every patient presenting sensory manifestations. Such vasculitic lesions are present in medium-sized arterioles and/or small vessels, and correspond mainly to 4 necrotizing vasculitis: panarteritis nodosa (PAN), microscopic polyangiitis (MPA), Churg and Strauss syndrome and Wegener granulomatosis. Microvasculitis should be added to these classical entities, because it corresponds to small vessel wall infiltration by inflammatory cells, as observed in PAN and MPA, but without any necrosis. Microvasculitis has to be differentiated from the inflammatory cell infiltrates surrounding small vessels. However, such perivascular inflammatory cell infiltrates enable the diagnosis of probable vasculitis when associated with clusters of neo-vessels, hemosiderin deposits, or a focal damage of nerve fibers. Grossly, one third of vasculitis diagnosis is confirmed on muscle fragments, a second third on nerve fragments, and the last third on both nerve and muscle fragments. Moreover, in the search for vasculitis, an unpredicted diagnosis of lymphoma or amyloidosis is occasionally established on the neuro-muscular biopsy. PMID- 17128152 TI - [MUC1 (EMA): A key molecule of carcinogenesis?]. AB - MUC1 is a large trans-membrane highly glycosylated mucin which is expressed at the apical pole of normal cells in glandular epithelia. MUC1 is implicated in many physiological mechanisms such as adhesion, development and differentiation. Also, MUC1 is frequently deregulated and over-expressed with a membrane circumferential and/or cytoplasmic expression. The intracellular tail of MUC1 is phosphorylated and can interact with many signalling proteins and transcriptional factors. Indeed, MUC1 can interact with B-catenin competitively for E-cadherin, thus destabilizing intercellular junctions and favouring metastatic dissemination. In carcinomas, the overexpression and membrane delocalization of MUC1 is associated with a worse prognosis and a shorter survival in breast, colon, kidney, prostate or gastro-intestinal cancers. MUC1 appears to be a novel therapeutic target for immunotherapy or anti-tumour vaccines. PMID- 17128153 TI - [Mixed cryoglobulinemia-associated membranoproliferative glomerulonephritis, disclosing gastric MALT lymphoma]. AB - We report a case of a mixed cryoglobulinemia-associated membranoproliferative glomerulonephritis, disclosing gastric lymphoma of MALT-type. In the reported case, glomerulonephritis was associated with joint and skin symptoms related to type II mixed cryoglobulinemia. MALT lymphoma diagnosis was made by gastric biopsy. Renal pathological examination showed relatively uncommon findings, ie mesangioproliferative glomerulonephritis lacking usual intraluminal thrombi. Although classical, renal manifestations of malignant lymphomas are uncommon, and rarely allow lymphoma diagnosis. The renal biological function improves as the lymphoma regresses with appropriated treatment. This case underlines the benefit of gastroscopy in cryoglobulinemia etiological research. PMID- 17128154 TI - [Gamna Gandy bodies: Features mimicking a mycotic disease. Two cases in the spleen]. AB - We report the clinical and histopathological features of 2 cases of Gamna-Gandy bodies located in the spleen. The diagnosis was made by the presence of circumscribed nodules containing dark brown rod-shaped and "bamboo-shaped" fibrotic structures. These structures were positive with Perls and von Kossa staining. Differential diagnosis includes filamentous mycoses such as mucormycosis and phaeohyphomycosis. Gamna-Gandy nodules can be isolated or these bodies can be observed associated with another lesion, particularly, a tumor. PMID- 17128155 TI - [The RNA revolution]. PMID- 17128156 TI - [Unusual tumor of the nasopharynx]. PMID- 17128157 TI - [A rare subcutaneous tumour]. PMID- 17128158 TI - [An esophageal allergy]. PMID- 17128159 TI - [Recurrence of a neck tumor]. PMID- 17128160 TI - [Unusual tumour of the parotid gland]. PMID- 17128161 TI - [An unusual intranodal lesion]. PMID- 17128162 TI - [A woman with an unusual bladder tumor]. PMID- 17128163 TI - [A rare localization of chest wall hydatidosis: Sternal hydatidosis]. PMID- 17128164 TI - [Malignant tumors of the penis in the Cameroons: epidemiological, clinical, pathological and therapeutical features]. PMID- 17128165 TI - [Massive liver cell necrosis and yellow fever]. PMID- 17128180 TI - [A merger opens the way...]. PMID- 17128181 TI - [Script concordance test: how to evaluate medical reasoning in uncertain situations]. PMID- 17128182 TI - [Dental extractions in children with congenital coagulation disorders: therapeutic protocol and results]. AB - INTRODUCTION: We report the results of our 10-year experience with a protocol of local hemostasis specifically elaborated for children with congenital bleeding disorders. MATERIAL AND METHOD: Sixty-four dental extractions procedures were performed in 37 children. Local hemostasis was achieved with the CO2 laser at the site of extraction previously filled with regenerated oxidized cellulose. The site of extraction was then protected with thermomolded splint. The replacement therapy (hemostasis factor concentrates, platelet concentrates) was coordinated by the hematologist and restricted as much as possible. RESULTS: 160 decidual teeth and 23 permanent teeth were extracted. None of the children with minor bleeding deficiency received replacement therapy. Ten post-operative bleedings were recorded but none was considered severe. DISCUSSION: This protocol characterized by the use of CO2 laser for the hemostasis gave satisfaction in children. The rare and minor post-operative bleedings could be explained by multiple extractions, local infection and/or children's anxiety. This strategy requires a close cooperation between the stomatologist and the hematologist in order to determine the optimal therapeutic management. PMID- 17128183 TI - [Double-barrel fibular free flap for mandibular reconstruction: beneficial alternative for dental implanted rehabilitation (5 cases)]. AB - INTRODUCTION: As major loss of mandibular bone stock requires a vascularized transfer, fibula free flap reconstruction is considered to be the best free flap for its length and reliability. Its main advantage is to accept dental implants. Single or double-barrel reconstruction can be performed. Double-barrel reconstruction is generally preferred because the bone superposition offers enough height to allow dental implants. MATERIAL AND METHODS: We reviewed five selected cases of double-barrel fibula free flap adaptive mandibular reconstruction performed among a series of 11 oral rehabilitations with planned dental implants, focusing on the technical aspects. RESULTS: Today, dental rehabilitation has been achieved in three double-barrel fibula flaps. Details are reported concerning the implant step. DISCUSSION: In this perspective, we discuss the choice of the reconstructive technique in order to obtain adequate bone height. Early in our experience and for different reasons discussed in the text, we used a single barrel fibula flap. This technique provided sufficient height in some cases, but had to be completed by bone grafts in few patients. Our experience illustrates the usefulness of the double barrel technique which provide definitive bone height sufficient for dental implants. The double barreled technique should be considered as the best solution. PMID- 17128185 TI - [Characteristic craniofacial features in a group of unilateral cleft lip and palate patients in Chile]. AB - INTRODUCTION: Craniofacial growth in cleft lip and palate patients is controversial. Structural alterations of the face in these patients could be due to the surgical treatment combined with the primary variations of the cranial base morphology. Our objective was to ana-lyze the morphology of the craniofacial skeleton in a group of unilateral cleft lip and palate Chilean patients. MATERIAL AND METHOD: We studied 43 non syndromic operated unilateral complete cleft lip and palate patients. Ages ranged from 10 to 18 years for both genders. This cohort was compared with a control group of 40 non cleft patients, matched for age and sex, using Delaire's structural and architectural craniofacial analysis on lateral radiographs and performed by the same operator. RESULTS: Cleft patients presented with significantly different configurations of the craniofacial skeleton, specifically a flatter basi-cranium, a reduced posterior maxillary height, a retrusive maxilla deficient in length with an associated crossbite. PMID- 17128186 TI - [Hypertrophic diseases of facial fatty tissue]. AB - Based on our own experience and a review of the literature, we des-cribe hypertrophic diseases concerning the fatty tissue of the face. Pathological patterns are numerous and polymporphous. Fatty localizations involving the face may be associated or not with specific histological presentation. PMID- 17128187 TI - [Paradontal plastic surgery: treatment of gingival recession using a gliding flap with a deep conjunctive graft]. PMID- 17128188 TI - [Infracondylar abscess formation: a rare complication of local anesthesia]. AB - INTRODUCTION: Ninety percent of oro-facial infections arise from a dental origin. The remaining 10% are the consequence of oro-pharyngeal, cutaneous or iatrogenic problems, such as in the present case. CASE REPORT: A 24-year-old patient consulted the emergency room because of a left mandibular swelling, accompagnied by trismus. Four days earlier, extraction of the 38 was performed under inferior alveolar nerve block anesthesia. A first drainage by vestibular approach was performed under general anesthesia. Because of the absence of improvement, a CT scan was performed and an abscess localized at the base of the condyle, surrounding the posterior margin of the mandible, very high above the lingula. Outcome was favourable after a second surgery. DISCUSSION: Formation of an abscess very high above the lingula, around the condylar neck is rarely reported in the literature. In this patient it was certainly a complication resulting from the injection of local anesthesic with a vasoconstrictor. CT-scan should be performed to guide diagnosis in the event of an unusual course after the first surgical procedure and an adequate antibiotic regimen. PMID- 17128189 TI - [Ganglioneuroma of the zygoma]. AB - INTRODUCTION: Ganglioneuroma is an uncommon benign tumor that arises from the sympathetic nervous system accounting for less than 1% of all soft-tissue neoplasms. CASE REPORT: We report the case of a 22 year-old man who presented a ganglioneuroma localized in the zygoma. DISCUSSION: Ganglioneuroma generally develop in the abdomen or thorax soft tissue. The occurrence of ganglioneuroma in the bone is exceptional. To date less than ten cases have been reported, six of which involved the mandible. We discuss the possible causes of tumor development at this site. PMID- 17128190 TI - [Post syphilitic oro-nasal communication]. AB - INTRODUCTION: Oro-nasal communication occurs due to a lack of bone and mucous membranes between the oral cavity and the nasal fossae, rarely subsequent to tertiary syphilis. CASE REPORT: A 27 year-old woman with a history of an untreated genital chancre which developed 7 years earlier, presented hard palate necrosis surrounded with granulation tissue. Syphilitic gum of the hard palate was diagnosed. After antibiotic treatment, the palatine lesion healed with persistence of an oro-nasal fistula. Physical examination of the oral cavity demonstrated median oro-nasal communication of the hard palate measuring 0,5 cm in diameter. HIV and syphilitic serologies were negative. The treatment consisted in surgical closure of the oro-nasal communication by the refection of nasal and oral plans. The follow-up was good. DISCUSSION: Oro-nasal communication due to syphilis is exceptional, of easy diagnosis and generally needs surgical treatment. PMID- 17128191 TI - [Lacrimal adenocarcinoma and anophtalmos]. AB - INTRODUCTION: The classical symptoms of orbital tumors are modified in the event of an anophthalmic socket. CASE REPORT: We report a case of primary malignant neoplasm of the lacrimal gland in an enucleated 70-year-old women. DISCUSSION: Patients with an anophthalmic orbit often present postenucleation socket syndrome: problems of tolerance and rocking of the ocular prothesis, enophthalmos, ptosis, lower eyelid ectropion. Development of a malignant tumor of the lacrimal gland is however exceptional. An oversized lacrimal gland is suggestive of tumor development. Rigorous clinical and radiological follow-up (CT, MRI) is essential. We discuss diagnostic and therapeutic aspects including the type of exenteration, radiotherapy, and reconstruction technique. PMID- 17128192 TI - [Chondroblastic osteosarcoma of the mandible: a case report with 14 years follow up]. AB - INTRODUCTION: Osteosarcoma, the second most frequent primary malignant bone tumor, is usually found in long bones: femur, tibia or humerus with only 6 to 7% of cases occurring in the jaws. Patients with osteosarcoma of the mandible usually complain of a swelling, which can be painful or not, accompanied by paresthesia of one of the trigeminal nerve branches in about 20% of cases. OBSERVATION: We report a case of chondroblastic osteosarcoma of the mandible affecting a 33-year-old woman with 14 years follow-up illustrating the difficulties of a rapid diagnosis. DISCUSSION: The prognosis of osteosarcoma of the mandible relies on its histological grade and the amount of time elapsing from diagnosis to treatment onset. The treatment of choice is radical surgery providing a 5-year survival rate up to 80%. The definitive surgical treatment is usually performed only after several interventions because operative-room pathology cannot be obtained due to the necessity of decalcification. In order to avoid multiple resections, an interim bridging with a reconstructive plate with a condylar head can be proposed before definitive graft reconstruction. The prognosis of osteosarcoma of the mandible is better than that of long bones. Chemotherapy or radiotherapy, which are very efficient for osteosarcoma in general, do not change the prognosis of osteosarcoma of the mandible. PMID- 17128193 TI - [Salivary duct carcinoma]. AB - INTRODUCTION: Salivary duct carcinoma is a very rare tumor of the salivary glands that has been principally reported in the parotid gland. It displays histologically a striking similarity to ductal carcinoma of the breast. OBSERVATION: We report the case of a salivary duct carcinoma arising in the parotid gland of a 62 year-old man. The fact that the tumor was confined to the parotid gland has allowed a radical treatment. After two year follow-up, there was no evidence of recurrence. DISCUSSION: This tumor is characterized clinically by local aggressiveness, frequent metastatic evolution and poor prognosis. This case report illustrates both the clinicopathological features and the prognostic factors of this entity. PMID- 17128194 TI - [Odontogenic myxoma of the maxillary]. AB - INTRODUCTION: Odontogenic myxoma is a rare benign tumor that arises from the mesenchymal portion of the tooth germ. It has a va-riable non-specific clinical and radiological appearance, and may be confused with other lesions such as ameloblastoma. CASE REPORT: We describe the case of a young, African male patient with a large odontogenic myxoma of the maxillary. The patient presented a painless firm swelling of the left maxillary. The radiograph showed a radiolucent lesion that was compatible with several diagnoses. DISCUSSION: The diagnosis was based on clinical, radiographic and histopathologic features. This tumor is locally aggressive, inducing important facial deformation and tooth displacement. Radical treatment with surgery or prosthesis is needed due to the high rate of recurrence. PMID- 17128195 TI - [Lingual nerve injury during removal of the lower third molar: importance of early intervention]. AB - INTRODUCTION: Lingual nerve injury is one of the most important complications after removal of a lower third molar. CASE REPORT: We report two cases of this complication in our experience. In the first case, lingual nerve injury was repaired immediately and the patient recovered total tongue sensitivity. In the second case, lingual nerve exploration was not undertaken and the patient developed complete tongue anesthesia. DISCUSSION: Opinions are quite different about the delay before exploration of an injured lingual nerve. However, absence of sensory improvement incited us to undertake surgical exploration of the nerve. Due to the degenerative lesion of the damaged nerve, timing of repair before three months seems preferable. In young patients, early repair, and good quality nerve suture without tissue loss improve sensitive recovery of the tongue. PMID- 17128196 TI - [Synovialosarcoma of the infraemporal fossa]. PMID- 17128197 TI - [Bisphosphonate-related bilateral mandibular osteochemonecrosis with pathological fracture of the horizontal ramus]. PMID- 17128198 TI - HLA-identical sibling allogeneic peripheral blood stem cell transplantation with reduced intensity conditioning compared to autologous peripheral blood stem cell transplantation for elderly patients with de novo acute myeloid leukemia. AB - We conducted a retrospective registry-based analysis to compare the outcome of 361 allogeneic human leukocyte antigen (HLA)-identical peripheral blood stem cell transplants (PBSCT) with reduced intensity conditioning (RIC) to that of 1369 autologous (auto) PBSCT in patients aged 50 years or older with de novo acute myeloid leukemia (AML), performed from 1997 until 2003 and reported to the European Group for Blood and Marrow Transplantation. Median age was 58 and 57 years in the RIC and auto groups, respectively. RIC patients had more advanced disease at the time of transplant. At a median follow-up of 24 months for RIC and 16 months for auto, multivariate analysis showed a lower risk for relapse (RR 0.77, P=0.013) without increased non-relapse mortality (NRM) in RIC patients (RR 1.26, P=0.28). Moreover, leukemia-free survival (RR 1.22, P=0.02) and overall survival (OS) (RR 1.32, P=0.005) were superior in the RIC group. In patients in 1st (CR), fewer relapses were counterbalanced by significantly increased NRM. Therefore, there was no survival advantage in this subgroup. In patients in 2nd or subsequent CR, LFS and OS were superior in the RIC group. RIC transplants show encouraging results in this older patient population with de novo AML. PMID- 17128199 TI - Angle-closure caused by an anterior segment membrane. PMID- 17128200 TI - Traumatic macular hole secondary to Nd:YAG laser. PMID- 17128201 TI - Scintigraphic evaluation for tear drainage, after dacryocystorhino- stomy, in relation to patient satisfaction. AB - AIM: To measure the degree of physiological patency of the tear drainage system using dacryoscintigraphy before and after external dacryocystorhinostomy (EDCR) in relation to patient complaints. METHODS: Prospective evaluation of 29 eyes of 24 patients with primary acquired nasolacrimal duct obstruction (PANDO) who underwent EDCR. The epiphora complaints were measured subjectively. Dacryoscintigraphy was performed in all eyes preoperatively, and was repeated 6 months after DCR together with the complaints score. For evaluation of dacryoscintigraphy, we determined T1 (percentage of administered dose still present after 1 min) and linear clearance rate (LCR), defined as: 100%(T1 T15)/T1) from the tracer disappearance curve. Dacryoscintigraphy findings were compared with the results of 20 eyes of normal volunteers without any symptoms. Scintigraphic findings were compared before and after the operation, with the change in patient complaints score as well as with normal values. RESULTS: Significant improvement occurred after the operation in the complaints score, T1 and LCR. However, despite absence of complaints (score=0) postoperative scintigraphic values were still abnormal as compared to normal individuals. CONCLUSION: Despite almost complete remission of epiphora complaints, DCR does not result in normalization of the tear drainage system. PMID- 17128202 TI - Intravitreal bevacizumab (Avastin) for choroidal metastasis secondary to breast carcinoma: short-term follow-up. PMID- 17128203 TI - Long-term results of Erbium YAG-laser-assisted deep sclerectomy. AB - PURPOSE: To evaluate the long-term results of Erbium YAG-laser-assisted deep sclerectomy (DS). In this procedure, the delicate dissection of a deep corneoscleral lamella is greatly simplified by using the Erbium YAG-laser. METHODS: Data of 14 consecutive patients (10 male, four female, age 67.7+/-10.4 years), who underwent surgery from 1999 to 2000 were analysed retrospectively. The procedure was begun as a standard DS. The deep corneoscleral lamella was dissected with a pulsed Erbium YAG-laser (energy: 40-100 mJ, frequency: 5-10 Hz). Schlemm's canal was unroofed and the lamella thinned until aqueous percolated continuously through the membrane. RESULTS: The mean follow-up time was 50.4+/ 6.8 months. The mean preoperative intraocular pressure (IOP) was 37.7+/-10.5 mmHg. The mean postoperative IOP was 16.1+/-3.9 mmHg at 1 month, 15.1+/-4.3 mmHg at 3 months, 16.4+/-4.5 mmHg at 12 months, and 17.6+/-8.7 mmHg at 50.5 months. The complete success rates (IOP< or =21 mmHg+IOP reduction > or =20% without glaucoma medication) were 83.3% at 3 months and 50% at 12 and 50.5 months. Rates for qualified success (IOP< or =21 mmHg+IOP reduction > or =20% with glaucoma medication) were 91.7% at 3 months, 92.9% at 12 months, and 78.6% at 50.5 months. The number of glaucoma medications was reduced from 3.07+/-0.92 preoperatively to 1.14+/-1.41 at 50.5 months. A single case of anterior-chamber penetration, requiring iridectomy, was the only intraoperative complication. CONCLUSIONS: Erbium YAG-laser-assisted DS has the advantage of a greatly simplified dissection, while offering a successful long-term IOP control comparable to conventional DS. PMID- 17128204 TI - Visual side effects after prolonged MRSA treatment. PMID- 17128205 TI - Cost-effectiveness of early detection and treatment of ocular hypertension and primary open-angle glaucoma by the ophthalmologist. AB - PURPOSE: To determine the most cost-effective case-finding strategy for the ophthalmologist to detect and treat ocular hypertension (OH) and primary open angle glaucoma (POAG) at an early stage to prevent blindness. DESIGN: A Markov cost-effectiveness simulation model. METHODS: Three case-finding strategies are analysed and compared. The simulated cohort consists of all initial patients of at least 40 years old visiting an ophthalmic practice. All patients undergo ophthalmoscopy, but tonometry is routinely performed to: (1) all initial patients, (2) high-risk patients only, or (3) no one. The population characteristics are based on data of 1000 initial patients. Transition probabilities are taken from the literature. The (direct) costs of diagnosis and treatment represent those for the Netherlands. The time-horizon of the model is 20 years. An annual discount rate of 4% is used. MAIN OUTCOME MEASURES: Costs, proportion of patients becoming blind, years of blindness. RESULTS: The costliest strategy (1) leads to least blindness. The incremental cost-effectiveness ratio, which shows extra costs per year of vision saved in comparison to the cheapest strategy (3), is lower for strategy (1) than for strategy (2). It amounts to euro1707, not including extra costs due to blindness (eg associated with the use of disability facilities). When such costs exceed euro1707 per patient per year, which is most likely, then strategy (1) becomes cost saving. CONCLUSION: It is most cost-effective to routinely perform tonometry to all initial ophthalmic patients to prevent blindness due to glaucoma. PMID- 17128207 TI - Reply to letter from Cooke et al: silicone oil migration causing increasing proptosis 13 years after retinal surgery. PMID- 17128208 TI - An unusual maculopathy after routine cataract surgery. AB - AIMS: To report the presentation of apparent acute and transient vitreomacular traction causing reduction in visual acuity following routine cataract surgery. METHODS: A case series describing three patients that developed acute and transient vitreomacular traction following routine cataract surgery by experienced surgeons. No patients had any significant past medical or ophthalmic history. The patients presented shortly after surgery with reduction in visual acuity and dramatic optical coherence tomography (OCT) images suggesting vitreomacular traction. RESULTS: All three patients experienced spontaneous resolution of symptoms and clinical signs within the first few weeks after surgery. Two patients experienced continued mild metamorphopsia. CONCLUSION: To our knowledge, this is the first documented report of such cases with associated confirmation by OCT imaging. PMID- 17128209 TI - Regulating the p53 pathway: in vitro hypotheses, in vivo veritas. AB - Mutations in TP53, the gene that encodes the tumour suppressor p53, are found in 50% of human cancers, and increased levels of its negative regulators MDM2 and MDM4 (also known as MDMX) downregulate p53 function in many of the rest. Understanding p53 regulation remains a crucial goal to design broadly applicable anticancer strategies based on this pathway. This Review of in vitro studies, human tumour data and recent mouse models shows that p53 post-translational modifications have modulatory roles, and MDM2 and MDM4 have more profound roles for regulating p53. Importantly, MDM4 emerges as an independent target for drug development, as its inactivation is crucial for full p53 activation. PMID- 17128210 TI - Gastrin - active participant or bystander in gastric carcinogenesis? AB - Gastrin is a pro-proliferative, anti-apoptotic hormone with a central role in acid secretion in the gastric mucosa and a long-standing association with malignant progression in transgenic mouse models. However, its exact role in human gastric malignancy requires further validation. Gastrin expression is tightly regulated by two closely associated hormones, somatostatin and gastrin releasing peptide, and aspects of their interaction may be deregulated during progression to gastric adenocarcinoma. Furthermore, agonists and antagonists of the receptors for all three hormones have shown modest clinical efficacy against gastric adenocarcinoma, which might provide useful information on the future combined use of these agents. PMID- 17128212 TI - Dentistry and domestic violence. PMID- 17128213 TI - Conflicting advice. PMID- 17128211 TI - The role of cytochrome P450 enzymes in endogenous signalling pathways and environmental carcinogenesis. AB - Some cytochrome P450 (CYP) heme-thiolate enzymes participate in the detoxication and, paradoxically, the formation of reactive intermediates of thousands of chemicals that can damage DNA, as well as lipids and proteins. CYP expression can also affect the production of molecules derived from arachidonic acid, and alters various downstream signal-transduction pathways. Such changes can be precursors to malignancy. Recent studies in mice have changed our perceptions about the function of CYP1 enzymes. We suggest a two-tiered system to predict an overall inter-individual risk of tumorigenesis based on DNA variants in certain 'early defence' CYP genes, combined with polymorphisms in various downstream target genes. PMID- 17128214 TI - Misplaced and inappropriate. PMID- 17128215 TI - Medico-legal dilemma. PMID- 17128216 TI - A unique opportunity? PMID- 17128217 TI - Reduction rationales. PMID- 17128218 TI - Confusing guidelines. PMID- 17128219 TI - Volunteer programme. PMID- 17128220 TI - Do no harm. PMID- 17128225 TI - Common antacids could help keep gingivitis at bay. PMID- 17128231 TI - Dental caries affects body weight, growth and quality of life in pre-school children. AB - The effect of a relatively common chronic disease, severe dental caries, affects young childrens' growth and well-being. Treating dental caries in pre-school children would increase growth rates and the quality of life of millions of children. Severe untreated dental caries is common in pre-school children in many countries. Children with severe caries weighed less than controls, and after treatment of decayed teeth there was more rapid weight gain and improvements in their quality of life. This may be due to dietary intake improving because pain affected the quantity and variety of food eaten, and second, chronic inflammation from caries related pulpitis and abscesses is known to suppress growth through a metabolic pathway and to reduce haemoglobin as a result of depressed erythrocyte production. PMID- 17128232 TI - Connecting implants to teeth. AB - Osseointegrated implants have proven to have a high degree of success in the edentulous arches. This success has been replicated in partially edentulous arches. When natural teeth are present, it can be tempting to fabricate restorations using support from both implants and natural teeth. The mechanism of attachment and the perceived problem of the differential support provided by the implant and the tooth have been discussed by many authors. PMID- 17128233 TI - The impact of clinical audit on antibiotic prescribing in general dental practice. AB - OBJECTIVE: To reduce the number of antibiotics inappropriately prescribed by general dental practitioners, and to increase overall prescription accuracy. DESIGN: A prospective clinical audit carried out between September and March of 2002-3 and 2003-4. SETTING: General dental practices in Eastern England. SUBJECTS AND METHODS: The pre-audit antibiotic prescribing practices of 212 general dental practitioners were recorded over an initial six week period. On each occasion this included which antibiotic had been chosen, together with its dose, frequency and duration, as well as the clinical condition and reason for which the prescription had been raised. When related to prophylaxis, the patient's medical history was also noted. Following education on contemporary prescribing guidelines, presentations which illustrated the practitioners' previous errors, and the agreement of standards to be achieved, the process was repeated for another six weeks, and the results compared. RESULTS: In the pre-audit period, 2,951 antibiotic prescriptions were issued, and during the audit this was reduced by 43.6% to 1,665. The majority were for therapeutic reasons, with only 10.5% and 13.6% for medical prophylaxis during the pre-audit and audit periods respectively. Over both periods, amoxicillin and metronidazole were the two most commonly prescribed antimicrobials (63.4% and 21.2% respectively). In the pre audit period, only 43% of all prescriptions were error free in dose, frequency, and/or duration of use, but this rose significantly to 78% during the audit. Equally, using contemporary published guidelines, out of all the prescriptions made in the pre-audit period, only 29.2% were deemed to be justified, as compared to 48.5% during the audit. CONCLUSIONS: Clinical audit, in conjunction with education, and prescribing guidelines can favourably change antibiotic prescribing patterns among general dental practitioners. PMID- 17128234 TI - An audit of the UK national cancer referral guidelines for suspected oral mucosal malignancy. AB - BACKGROUND AND OBJECTIVE: This study aimed to assess the effectiveness of the UK National Guidelines for identifying patients with potentially malignant oral disease which were introduced in 2000. DESIGN: Retrospective audit. SETTING: The oral medicine unit in a university teaching hospital in London. METHODS: All new referrals over a one year period were retrospectively reviewed in a departmental audit to evaluate guideline effectiveness. Reasons for referral and final diagnosis were compared in a randomly selected sub-population. RESULTS: Four hundred and eighty-seven of 901 new patients referred were classified as having potentially malignant disease from the referral letter. In a randomly selected subgroup of 241 patients, 18 actually had malignant (8) or dysplastic lesions (10). Of 75 patients referred with a persistent oral ulcer, only nine were actually malignant or dysplastic. Eight of 116 patients referred with a white patch and none with red patches were found to have dysplastic or malignant lesions. The criteria failed to identify three carcinomas and two severely dysplastic lesions (15% of the malignant or dysplastic lesions). All of the latter had been referred by primary care physicians with orofacial pain of unknown cause. CONCLUSIONS: UK National Guidelines discriminate poorly between potentially malignant and other oral mucosal disease. PMID- 17128240 TI - Changes in the levels of knowledge and attitudes of dental hospital staff about domestic violence following attendance at an awareness raising seminar. AB - OBJECTIVES: To assess changes in dental team knowledge and awareness about domestic violence following attendance at a brief training intervention. MATERIALS AND METHODS: Seventy-five dental hospital staff at the University of Manchester Dental Hospital attended a two-hour session delivered by Manchester's City Wide NHS Domestic Violence Project manager. Participants completed identical questionnaires immediately before and after the session. Paired t-tests were conducted to determine changes in responses to individual questions before and after training. Independent sample t-tests were also conducted to compare mean responses by sex. RESULTS: Statistically significant improvements were seen in 50% of attitude questions and 100% of knowledge questions. After training, there was improved recognition that interpersonal violence was a health issue and that the dental profession should be more involved in identification of abuse. There was also improved comfort asking about abuse and respondents were less afraid of offending the patient and less likely to blame the victims for being abused. Statistically significant sex differences at baseline were also seen for several items: females obtaining more correct answers at baseline than males. CONCLUSIONS: Brief domestic violence training interventions can be effective in improving knowledge and attitudes amongst a dental team but could lead to false confidence in staff and should be followed by in-depth practical training and the development of appropriate processes for dealing with abuse victims. PMID- 17128241 TI - Educational needs and employment status of Scottish dental nurses. AB - OBJECTIVE: To investigate the educational needs and employment status of dental nurses in Scotland. SUBJECTS: One thousand, seven hundred and thirty dental nurses with postal addresses in Scotland. DESIGN: Structured questionnaire. RESULTS: A 65% response rate was achieved following three mailings. The majority of respondents were employed in the central belt of Scotland although comparatively, substantial numbers were also recorded as being employed in Highland and Grampian regions. A total of 70% of dental nurses stated they were in full-time employment, with 74% of all participants reporting they were qualified. Of those who were unqualified, 17% (n = 191) were not in the process of formal training which may mean they are unable to work in the future, unless they conform to the GDC 'grandparenting' requirements. Induction training had only been received by 34% (n = 377) of this group, and a mere 21% (n = 234) reported regular CPD involvement. Of the 75% (n = 832) who responded to a question on funding for CPD, only 50% (412) received financial assistance. Geographical location for many nurses generated a number of difficulties in accessing education, and the possibility of distance or e-learning was requested by 65% of the sample. CONCLUSIONS: This study demonstrates clearly that there are many well-qualified dental nurses in employment in Scotland. However, unless structured career pathways are developed, access to education is simplified, and the problem of poor remuneration is addressed, then foreseeable problems may arise where there is insufficient enticement to encourage people to join what is an already underpopulated profession. PMID- 17128255 TI - Polyadenylation factor CPSF-73 is the pre-mRNA 3'-end-processing endonuclease. AB - Most eukaryotic messenger RNA precursors (pre-mRNAs) undergo extensive maturational processing, including cleavage and polyadenylation at the 3'-end. Despite the characterization of many proteins that are required for the cleavage reaction, the identity of the endonuclease is not known. Recent analyses indicated that the 73-kDa subunit of cleavage and polyadenylation specificity factor (CPSF-73) might be the endonuclease for this and related reactions, although no direct data confirmed this. Here we report the crystal structures of human CPSF-73 at 2.1 A resolution, complexed with zinc ions and a sulphate that might mimic the phosphate group of the substrate, and the related yeast protein CPSF-100 (Ydh1) at 2.5 A resolution. Both CPSF-73 and CPSF-100 contain two domains, a metallo-beta-lactamase domain and a novel beta-CASP (named for metallo beta-lactamase, CPSF, Artemis, Snm1, Pso2) domain. The active site of CPSF-73, with two zinc ions, is located at the interface of the two domains. Purified recombinant CPSF-73 possesses RNA endonuclease activity, and mutations that disrupt zinc binding in the active site abolish this activity. Our studies provide the first direct experimental evidence that CPSF-73 is the pre-mRNA 3' end-processing endonuclease. PMID- 17128256 TI - From a two-dimensional chemical pattern to a three-dimensional topology through selective inversion of a liquid-liquid bilayer. AB - Soft organic surfaces with more and more complex topologies are required daily to engineer appropriate microstructures for many different applications such as DNA array technology, biological optics for advanced photonic systems and microfluidics. Complementarily to conventional lithographic processes, several pioneering methods have been developed recently, by controlling phase separation of polymer blends, spinodal decomposition of homopolymers or by using the action of additional external forces driving diverse instabilities. Here we present a method that not only provides original concepts towards the three-dimensional (3D) structuring of liquids, on the basis of the synergistic effects of molecular diffusion and confined nucleation, but also suggests original solutions for the transport, mixing and filtering of small volumes of liquid. Through the intrinsic destabilization of a liquid-liquid bilayer, the 2D pattern of a chemically structured surface with 'hydrophilic' and 'hydrophobic' domains is transferred to a solid/liquid interface as a 3D topography with either 'positive' or 'negative' replication. This easy-to-use process has potential applications in various technological realms requiring a specific topography at interfaces such as microfluidics or biosensors. PMID- 17128257 TI - Three-dimensional silicon inverse photonic quasicrystals for infrared wavelengths. AB - Quasicrystals are a class of lattices characterized by a lack of translational symmetry. Nevertheless, the points of the lattice are deterministically arranged, obeying rotational symmetry. Thus, we expect properties that are different from both crystals and glasses. Indeed, naturally occurring electronic quasicrystals (for example, AlPdMn metal alloys) show peculiar electronic, vibrational and physico-chemical properties. Regarding artificial quasicrystals for electromagnetic waves, three-dimensional (3D) structures have recently been realized at GHz frequencies and 2D structures have been reported for the near infrared region. Here, we report on the first fabrication and characterization of 3D quasicrystals for infrared frequencies. Using direct laser writing combined with a silicon inversion procedure, we achieve high-quality silicon inverse icosahedral structures. Both polymeric and silicon quasicrystals are characterized by means of electron microscopy and visible-light Laue diffraction. The diffraction patterns of structures with a local five-fold real-space symmetry axis reveal a ten-fold symmetry as required by theory for 3D structures. PMID- 17128258 TI - Shape-engineerable and highly densely packed single-walled carbon nanotubes and their application as super-capacitor electrodes. AB - We present a rational and general method to fabricate a high-densely packed and aligned single-walled carbon-nanotube (SWNT) material by using the zipping effect of liquids to draw tubes together. This bulk carbon-nanotube material retains the intrinsic properties of individual SWNTs, such as high surface area, flexibility and electrical conductivity. By controlling the fabrication process, it is possible to fabricate a wide range of solids in numerous shapes and structures. This dense SWNT material is advantageous for numerous applications, and here we demonstrate its use as flexible heaters as well as supercapacitor electrodes for compact energy-storage devices. PMID- 17128259 TI - Precision control of single-molecule electrical junctions. AB - There is much discussion of molecules as components for future electronic devices. However, the contacts, the local environment and the temperature can all affect their electrical properties. This sensitivity, particularly at the single molecule level, may limit the use of molecules as active electrical components, and therefore it is important to design and evaluate molecular junctions with a robust and stable electrical response over a wide range of junction configurations and temperatures. Here we report an approach to monitor the electrical properties of single-molecule junctions, which involves precise control of the contact spacing and tilt angle of the molecule. Comparison with ab initio transport calculations shows that the tilt-angle dependence of the electrical conductance is a sensitive spectroscopic probe, providing information about the position of the Fermi energy. It is also shown that the electrical properties of flexible molecules are dependent on temperature, whereas those of molecules designed for their rigidity are not. PMID- 17128260 TI - Multicomponent semiconducting polymer systems with low crystallization-induced percolation threshold. AB - Blends and other multicomponent systems are used in various polymer applications to meet multiple requirements that cannot be fulfilled by a single material. In polymer optoelectronic devices it is often desirable to combine the semiconducting properties of the conjugated species with the excellent mechanical properties of certain commodity polymers. Here we investigate bicomponent blends comprising semicrystalline regioregular poly(3-hexylthiophene) and selected semicrystalline commodity polymers, and show that, owing to a highly favourable, crystallization-induced phase segregation of the two components, during which the semiconductor is predominantly expelled to the surfaces of cast films, we can obtain vertically stratified structures in a one-step process. Incorporating these as active layers in polymer field-effect transistors, we find that the concentration of the semiconductor can be reduced to values as low as 3 wt% without any degradation in device performance. This is in stark contrast to blends containing an amorphous insulating polymer, for which significant reduction in electrical performance was reported. Crystalline crystalline/semiconducting-insulating multicomponent systems offer expanded flexibility for realizing high-performance semiconducting architectures at drastically reduced materials cost with improved mechanical properties and environmental stability, without the need to design all performance requirements into the active semiconducting polymer itself. PMID- 17128261 TI - Environmentally degradable, high-performance thermoplastics from phenolic phytomonomers. AB - Aliphatic polyesters, such as poly(lactic acid), which degrade by hydrolysis, from naturally occurring molecules form the main components of biodegradable plastics. However, these polyesters have become substitutes for only a small percentage of the currently used plastic materials because of their poor thermal and mechanical properties. Polymers that degrade into natural molecules and have a performance closer to that of engineering plastics would be highly desirable. Although the use of a high-strength filler such as a bacterial cellulose or modified lignin greatly increases the plastic properties, it is the matrix polymer that determines the intrinsic properties of the composite. The introduction of an aromatic component into the thermoplastic polymer backbone is an efficient method to intrinsically improve the material performance. Here, we report the preparation of environmentally degradable, liquid crystalline, wholly aromatic polyesters. The polyesters were derived from polymerizable plant-derived chemicals--in other words, 'phytomonomers' that are widely present as lignin biosynthetic precursors. The mechanical performance of these materials surpasses that of current biodegradable plastics, with a mechanical strength, sigma, of 63 MPa, a Young's modulus, E, of 16 GPa, and a maximum softening temperature of 169 degrees C. On light irradiation, their mechanical properties improved further and the rate of hydrolysis accelerated. PMID- 17128262 TI - Post-translational enzyme activation in an animal via optimized conditional protein splicing. AB - Control over the timing, location and level of protein activity in vivo is crucial to understanding biological function. Living systems are able to respond to external and internal stimuli rapidly and in a graded fashion by maintaining a pool of proteins whose activities are altered through post-translational modifications. Here we show that the process of protein trans-splicing can be used to modulate enzymatic activity both in cultured cells and in Drosophila melanogaster. We used an optimized conditional protein splicing system to rapidly trigger the in vivo ligation of two inactive fragments of firefly luciferase in a tunable manner. This technique provides a means of controlling enzymatic function with greater speed and precision than with standard genetic techniques and is a useful tool for probing biological processes. PMID- 17128263 TI - Phospholipase Cgamma1 negatively regulates growth hormone signalling by forming a ternary complex with Jak2 and protein tyrosine phosphatase-1B. AB - Growth hormone binds to its membrane receptor (GHR), whereby it regulates many cellular functions, including proliferation, differentiation and chemotaxis. However, although the activation of growth hormone-mediated signalling is well understood, the precise mechanism responsible for its regulation has not been elucidated. Here, we demonstrate that phospholipase Cgamma1 (PLCgamma1) modulates the action of growth hormone-mediated signalling by interacting with tyrosine kinase Jak2 (janus kinase 2) in a growth hormone-dependent manner. In the absence of PLCgamma1 (PLCgamma1(-/-)), growth hormone-induced JAK2 and STAT5 phosphorylation significantly increased in mouse embryonic fibroblasts (MEFs). Furthermore, the re-expression of PLCgamma1 reduced growth hormone-induced Jak2 activation. Growth hormone-induced Jak2 phosphorylation was enhanced by siRNA specific knockdown of PLCgamma1. Interestingly, PLCgamma1 physically linked Jak2 and protein tyrosine phosphatase-1B (PTP-1B) by binding to both using different domains, and this process was implicated in the modulation of cytokine signalling through Jak2. In addition, in PLCgamma1(-/-) MEFs, growth hormone-dependent c-Fos activation was upregulated and growth hormone-induced proliferation was potentiated. These results suggest that PLCgamma1 has a key function in the regulation of growth hormone-mediated signalling by negatively regulating Jak2 activation. PMID- 17128264 TI - Tumour-mediated upregulation of chemoattractants and recruitment of myeloid cells predetermines lung metastasis. AB - Primary tumours influence the environment in the lungs before metastasis. However, the mechanism of metastasis is not well understood. Here, we show that the inflammatory chemoattractants S100A8 and S100A9, whose expression is induced by distant primary tumours, attract Mac 1 (macrophage antigen 1)(+)-myeloid cells in the premetastatic lung. In addition, tumour cells use this mechanism, through activation of the mitogen-activated protein kinase (MAPK) p38, to acquire migration activity with pseudopodia for invasion (invadopodia). The expression of S100A8 and S100A9 was eliminated in lung Mac 1(+)-myeloid cells and endothelial cells deprived of soluble factors, such as vascular endothelial growth factor A (VEGF-A), tumour necrosis factor alpha (TNFalpha) and transforming growth factor beta (TGFbeta) both in vitro and in vivo. Neutralizing anti-S100A8 and anti S100A9 antibodies blocked the morphological changes and migration of tumour cells and Mac 1(+)-myeloid cells. Thus, the S100A8 and S100A9 pathway may be common to both myeloid cell recruitment and tumour-cell invasion. PMID- 17128265 TI - Maintenance of colonic homeostasis by distinctive apical TLR9 signalling in intestinal epithelial cells. AB - The mechanisms by which commensal bacteria suppress inflammatory signalling in the gut are still unclear. Here, we present a cellular mechanism whereby the polarity of intestinal epithelial cells (IECs) has a major role in colonic homeostasis. TLR9 activation through apical and basolateral surface domains have distinct transcriptional responses, evident by NF-kappaB activation and cDNA microarray analysis. Whereas basolateral TLR9 signals IkappaBalpha degradation and activation of the NF-kappaB pathway, apical TLR9 stimulation invokes a unique response in which ubiquitinated IkappaB accumulates in the cytoplasm preventing NF-kappaB activation. Furthermore, apical TLR9 stimulation confers intracellular tolerance to subsequent TLR challenges. IECs in TLR9-deficient mice, when compared with wild-type and TLR2-deficient mice, display a lower NF-kappaB activation threshold and these mice are highly susceptible to experimental colitis. Our data provide a case for organ-specific innate immunity in which TLR expression in polarized IECs has uniquely evolved to maintain colonic homeostasis and regulate tolerance and inflammation. PMID- 17128266 TI - PKC-1 regulates secretion of neuropeptides. AB - The secretion of neurotransmitters and neuropeptides is mediated by distinct organelles-synaptic vesicles (SVs) and dense-core vesicles (DCVs), respectively. Relatively little is known about the factors that differentially regulate SV and DCV secretion. Here we show that protein kinase C-1 (PKC-1), which is most similar to the vertebrate PKC eta and epsilon isoforms, regulates exocytosis of DCVs in Caenorhabditis elegans motor neurons. Mutants lacking PCK-1 activity had delayed paralysis induced by the acetylcholinesterase inhibitor aldicarb, whereas mutants with increased PKC-1 activity had more rapid aldicarb-induced paralysis. Imaging and electrophysiological assays indicated that SV release occurred normally in pkc-1 mutants. By contrast, genetic analysis of aldicarb responses and imaging of fluorescently tagged neuropeptides indicated that mutants lacking PKC-1 had reduced neuropeptide secretion. Similar neuropeptide secretion defects were found in mutants lacking unc-31 (encoding the protein CAPS) or unc-13 (encoding Munc13). These results suggest that PKC-1 selectively regulates DCV release from neurons. PMID- 17128267 TI - Fast manipulation of cellular cAMP level by light in vivo. AB - The flagellate Euglena gracilis contains a photoactivated adenylyl cyclase (PAC), consisting of the flavoproteins PACalpha and PACbeta. Here we report functional expression of PACs in Xenopus laevis oocytes, HEK293 cells and in Drosophila melanogaster, where neuronal expression yields light-induced changes in behavior. The activity of PACs is strongly and reversibly enhanced by blue light, providing a powerful tool for light-induced manipulation of cAMP in animal cells. PMID- 17128268 TI - Transposon-aided capture (TRACA) of plasmids resident in the human gut mobile metagenome. AB - The bacterial community of the human gut is a complex ecosystem composed of >1,000 species, the majority of which are Gram positive and uncultured. To study plasmids resident within this community, we developed a culture-independent transposon aided capture method (TRACA), which does not rely on any plasmid encoded traits. TRACA facilitated acquisition of plasmids from metagenomic DNA extracts, and subsequent maintenance and selection in an Escherichia coli host. We confirmed the presence of the transposon in captured plasmids and demonstrate that these plasmids are mainly of a Gram-positive origin. Sequencing of plasmids designated pTRACA10 (7 kb) and pTRACA17 (2.7 kb) revealed genes involved in plasmid mobilization and replication. From the homologies of these genes we conclude that pTRACA17 originates from a Gram-positive host belonging to the Firmicute division. pTRACA10 had two additional open reading frames with similarity to a conserved hypothetical protein and phosphoesterase or phosphohydrolase enzymes (Clusters of Orthologous Groups number 4186). Both plasmids lacked any conventional selectable markers. PMID- 17128269 TI - Regulation of osteoclast differentiation and function by the CaMK-CREB pathway. AB - Calcium (Ca(2+)) signaling is essential for a variety of cellular responses and higher biological functions. Ca(2+)/calmodulin-dependent kinases (CaMKs) and the phosphatase calcineurin activate distinct downstream pathways that are mediated by the transcription factors cAMP response element (CRE)-binding protein (CREB) and nuclear factor of activated T cells (NFAT), respectively. The importance of the calcineurin-NFAT pathway in bone metabolism has been demonstrated in osteoclasts, osteoblasts and chondrocytes. However, the contribution of the CaMK CREB pathway is poorly understood, partly because of the difficulty of dissecting the functions of homologous family members. Here we show that the CaMKIV-CREB pathway is crucial for osteoclast differentiation and function. Pharmacological inhibition of CaMKs as well as the genetic ablation of Camk4 reduced CREB phosphorylation and downregulated the expression of c-Fos, which is required for the induction of NFATc1 (the master transcription factor for osteoclastogenesis) that is activated by receptor activator of NF-kappaB ligand (RANKL). Furthermore, CREB together with NFATc1 induced the expression of specific genes expressed by differentiated osteoclasts. Thus, the CaMK-CREB pathway biphasically functions to regulate the transcriptional program of osteoclastic bone resorption, by not only enhancing induction of NFATc1 but also facilitating NFATc1-dependent gene regulation once its expression is induced. This provides a molecular basis for a new therapeutic strategy for bone diseases. PMID- 17128270 TI - v-ATPase V0 subunit d2-deficient mice exhibit impaired osteoclast fusion and increased bone formation. AB - Matrix-producing osteoblasts and bone-resorbing osteoclasts maintain bone homeostasis. Osteoclasts are multinucleated, giant cells of hematopoietic origin formed by the fusion of mononuclear pre-osteoclasts derived from myeloid cells. Fusion-mediated giant cell formation is critical for osteoclast maturation; without it, bone resorption is inefficient. To understand how osteoclasts differ from other myeloid lineage cells, we previously compared global mRNA expression patterns in these cells and identified genes of unknown function predominantly expressed in osteoclasts, one of which is the d2 isoform of vacuolar (H(+)) ATPase (v-ATPase) V(0) domain (Atp6v0d2). Here we show that inactivation of Atp6v0d2 in mice results in markedly increased bone mass due to defective osteoclasts and enhanced bone formation. Atp6v0d2 deficiency did not affect differentiation or the v-ATPase activity of osteoclasts. Rather, Atp6v0d2 was required for efficient pre-osteoclast fusion. Increased bone formation was probably due to osteoblast-extrinsic factors, as Atp6v02 was not expressed in osteoblasts and their differentiation ex vivo was not altered in the absence of Atp6v02. Our results identify Atp6v0d2 as a regulator of osteoclast fusion and bone formation, and provide genetic data showing that it is possible to simultaneously inhibit osteoclast maturation and stimulate bone formation by therapeutically targeting the function of a single gene. PMID- 17128271 TI - Evidence that microRNAs are associated with translating messenger RNAs in human cells. AB - MicroRNAs (miRNAs) regulate gene expression post-transcriptionally by binding the 3' untranslated regions of target mRNAs. We examined the subcellular distribution of three miRNAs in exponentially growing HeLa cells and found that the vast majority are associated with mRNAs in polysomes. Several lines of evidence indicate that most of these mRNAs, including a known miRNA-regulated target (KRAS mRNA), are actively being translated. PMID- 17128272 TI - Human let-7a miRNA blocks protein production on actively translating polyribosomes. AB - MicroRNAs (miRNAs) regulate gene expression at a post-transcriptional level through base-pairing to 3' untranslated regions (UTRs) of messenger RNAs. The mechanism by which human let-7a miRNA regulates mRNA translation was examined in HeLa cells expressing reporter mRNAs containing the Caenorhabditis elegans lin-41 3' UTR. let-7a miRNA strongly repressed translation, yet the majority of control and lin-41-bearing RNAs sedimented with polyribosomes in sucrose gradients; these polyribosomes, together with let-7a miRNA and the miRISC protein AGO, were released from those structures by puromycin. RNA containing the lin-41 3' UTR and an iron response element in the 5' UTR sedimented with polysomes when cells were incubated with iron, but showed ribosome run-off when the iron was chelated. These data indicate that let-7a miRNA inhibits actively translating polyribosomes. Nascent polypeptide coimmunoprecipitation experiments further suggest that let-7a miRNA interferes with the accumulation of growing polypeptides. PMID- 17128273 TI - Glycan optimization of a human monoclonal antibody in the aquatic plant Lemna minor. AB - N-glycosylation is critical to the function of monoclonal antibodies (mAbs) and distinguishes various systems used for their production. We expressed human mAbs in the small aquatic plant Lemna minor, which offers several advantages for manufacturing therapeutic proteins free of zoonotic pathogens. Glycosylation of a mAb against human CD30 was optimized by co-expressing the heavy and light chains of the mAb with an RNA interference construct targeting expression of the endogenous alpha-1,3-fucosyltransferase and beta-1,2-xylosyltransferase genes. The resultant mAbs contained a single major N-glycan species without detectable plant-specific N-glycans and had better antibody-dependent cell-mediated cytotoxicity and effector cell receptor binding activities than mAbs expressed in cultured Chinese hamster ovary (CHO) cells. PMID- 17128274 TI - Wnt-beta-catenin signaling initiates taste papilla development. AB - Fungiform taste papillae form a regular array on the dorsal tongue. Taste buds arise from papilla epithelium and, unusually for epithelial derivatives, synapse with neurons, release neurotransmitters and generate receptor and action potentials. Despite the importance of taste as one of our five senses, genetic analyses of taste papilla and bud development are lacking. We demonstrate that Wnt-beta-catenin signaling is activated in developing fungiform placodes and taste bud cells. A dominant stabilizing mutation of epithelial beta-catenin causes massive overproduction of enlarged fungiform papillae and taste buds. Likewise, genetic deletion of epithelial beta-catenin or inhibition of Wnt-beta catenin signaling by ectopic dickkopf1 (Dkk1) blocks initiation of fungiform papilla morphogenesis. Ectopic papillae are innervated in the stabilizing beta catenin mutant, whereas ectopic Dkk1 causes absence of lingual epithelial innervation. Thus, Wnt-beta-catenin signaling is critical for fungiform papilla and taste bud development. Altered regulation of this pathway may underlie evolutionary changes in taste papilla patterning. PMID- 17128275 TI - Genome-wide analysis of Arabidopsis thaliana DNA methylation uncovers an interdependence between methylation and transcription. AB - Cytosine methylation, a common form of DNA modification that antagonizes transcription, is found at transposons and repeats in vertebrates, plants and fungi. Here we have mapped DNA methylation in the entire Arabidopsis thaliana genome at high resolution. DNA methylation covers transposons and is present within a large fraction of A. thaliana genes. Methylation within genes is conspicuously biased away from gene ends, suggesting a dependence on RNA polymerase transit. Genic methylation is strongly influenced by transcription: moderately transcribed genes are most likely to be methylated, whereas genes at either extreme are least likely. In turn, transcription is influenced by methylation: short methylated genes are poorly expressed, and loss of methylation in the body of a gene leads to enhanced transcription. Our results indicate that genic transcription and DNA methylation are closely interwoven processes. PMID- 17128276 TI - The kinases aurora B and mTOR regulate the G1-S cell cycle progression of T lymphocytes. AB - CD28-deficient T cells arrest at the G1-S transition of the cell cycle. Here we show that this is controlled by the kinase aurora B, which exists in a complex with survivin and mammalian target of rapamycin (mTOR). Expression of aurora B in Cd28-/- T cells augmented phosphorylation of mTOR substrates, expression of cyclin A, hyperphosphorylation of retinoblastoma protein and activation of cyclin dependent kinases 1 and 2 and promoted cell cycle progression. Interleukin 2 enhanced aurora B activity, and inactive aurora B prevented interleukin 2-induced proliferation. Moreover, expression of aurora B restored Cd28-/- T cell proliferation and promoted inflammation in vivo. These data identify aurora B, along with survivin and mTOR, as a regulator of the G1-S checkpoint in T cells. PMID- 17128278 TI - Sound advice on science. PMID- 17128277 TI - In the absence of aminopeptidase ERAAP, MHC class I molecules present many unstable and highly immunogenic peptides. AB - Immunosurveillance by cytotoxic T cells requires that cells generate a diverse spectrum of peptides for presentation by major histocompatibility complex (MHC) class I molecules. Those peptides are generated by proteolysis, which begins in the cytoplasm and continues in the endoplasmic reticulum by the unique aminopeptidase ERAAP. The overall extent to which trimming by ERAAP modifies the peptide pool and the immunological consequences of ERAAP deficiency are unknown. Here we show that the peptide-MHC repertoire of ERAAP-deficient mice was missing many peptides. Furthermore, ERAAP-deficient cells presented many unstable and structurally unique peptide-MHC complexes, which elicited potent CD8+ T cell and B cell responses. Thus, ERAAP is a 'quintessential editor' of the peptide-MHC repertoire and, paradoxically, its absence enhances immunogenicity. PMID- 17128279 TI - High times for memory: cannabis disrupts temporal coordination among hippocampal neurons. AB - Exogenous cannabinoid receptor agonists impair hippocampus-dependent learning and decrease the power of hippocampal electroencephalographic activity. A new paper shows that cannabinoids desynchronize neuronal assemblies without affecting average firing rates, and that this effect correlates with memory deficits in individuals. PMID- 17128280 TI - Triggering the brain's pathology sensor. AB - Microglia, the brain's intrinsic immune cells, rapidly sense brain injury and help clear cellular debris. Haynes et al. now show that P2Y12 receptors are critical for activating microglia and directing them to the site of injury. PMID- 17128281 TI - Opioid, cheating on its receptors, exacerbates pain. AB - Opioids are commonly thought of as compounds that alleviate pain. A new study finds that elevated levels of the opioid dynorphin can unexpectedly activate bradykinin receptors, contributing to the maintenance of neuropathic pain. PMID- 17128283 TI - Stabilizing the visual world. AB - Sensory systems are thought to use an internal copy of motor commands to determine which sensations are caused by the self versus the world. A new paper proves that an identified pathway performs this function for eye movements. PMID- 17128284 TI - Structural and mechanistic insights into hepatitis C viral translation initiation. AB - Hepatitis C virus uses an internal ribosome entry site (IRES) to control viral protein synthesis by directly recruiting ribosomes to the translation-start site in the viral mRNA. Structural insights coupled with biochemical studies have revealed that the IRES substitutes for the activities of translation-initiation factors by binding and inducing conformational changes in the 40S ribosomal subunit. Direct interactions of the IRES with initiation factor eIF3 are also crucial for efficient translation initiation, providing clues to the role of eIF3 in protein synthesis. PMID- 17128285 TI - Little or no ability of obestatin to interact with ghrelin or modify motility in the rat gastrointestinal tract. AB - BACKGROUND AND PURPOSE: Obestatin, encoded by the ghrelin gene may inhibit gastrointestinal (GI) motility. This activity was re-investigated. EXPERIMENTAL APPROACH: Rat GI motility was studied in vitro (jejunum contractility and cholinergically-mediated contractions of forestomach evoked by electrical field stimulation; EFS) and in vivo (gastric emptying and intestinal myoelectrical activity). Ghrelin receptor function was studied using a GTPgammaS assay and transfected cells. KEY RESULTS: Contractions of the jejunum or forestomach were unaffected by obestatin 100 nM or 0.01-1000 nM, respectively (P>0.05 each; n=4 18). Obestatin (0.1-1 nM) reduced the ability of ghrelin 1 microM to facilitate EFS-evoked contractions of the stomach (increases were 42.7+/-7.8% and 21.2+/-5.0 % in the absence and presence of obestatin 1 nM; P<0.05; n=12); higher concentrations (10-1000 nM) tended to reduce the response to ghrelin but changes were not statistically significant. Similar concentrations of obestatin did not significantly reduce a facilitation of contractions caused by the 5-HT(4) receptor agonist prucalopride, although an inhibitory trend occurred at the higher concentrations (increases were 69.3+/-14.0% and 42.6+/-8.7% in the absence and presence of 1000 nM obestatin; n=10). Obestatin (up to 10 microM) did not modulate recombinant ghrelin receptor function. Ghrelin increased gastric emptying and reduced MMC cycle time; obestatin (1000 and 30,000 pmol kg(-1) min( 1)) had no effects. Obestatin (2500 pmol kg(-1) min(-1), starting 10 min before ghrelin) did not prevent the ability of ghrelin (500 pmol kg(-1) min(-1)) to shorten MMC cycle time. CONCLUSIONS AND IMPLICATIONS: Obestatin has little ability to modulate rat GI motility. PMID- 17128286 TI - Elevated pressure selectively blunts flow-evoked vasodilatation in rat mesenteric small arteries. AB - BACKGROUND AND PURPOSE: The present study investigated mechanisms underlying impaired endothelium-dependent vasodilatation elicited by elevating the intraluminal pressure in rat mesenteric small arteries. EXPERIMENTAL APPROACH: Arterial segments (internal diameter 316+/-2 microm, n=86) were mounted in a pressure myograph. The effect of elevating pressure from 50 to 120 mmHg for 1 h before resetting it to 50 mmHg was studied on endothelium-dependent vasodilatation. KEY RESULTS: In arteries constricted with U46619 in the presence of indomethacin, shear stress generated by flow, evoked vasodilatation that was abolished by an inhibitor of nitric oxide (NO) synthase, asymmetric dimethylarginine (1 mM), whereas acetylcholine-induced vasodilatation was unchanged. After elevation of intraluminal pressure for 1 h and then resetting it to 50 mmHg, vasodilatation induced by shear stress and the NO donor, S nitrosopenicillamine was inhibited, while vasodilatation induced by a guanylyl cyclase activator, BAY 412272, and acetylcholine was unaltered. Superoxide levels sensitive to polyethylene glycol superoxide dismutase were increased in segments exposed to elevated pressure. A superoxide scavenger, tempol (300 microM), a general endothelin receptor antagonist, SB 217242 and the selective ET(A) receptor antagonist, BQ 123 preserved shear stress-evoked vasodilatation. CONCLUSIONS AND IMPLICATIONS: The present study shows that transient exposure to an elevated intraluminal pressure selectively inhibits flow-evoked NO-mediated vasodilatation, probably through activation of endothelin receptors and increased formation of superoxide. In contrast, elevation of pressure did not affect the acetylcholine-evoked endothelium-derived hyperpolarizing factor type vasodilatation in mesenteric small arteries. PMID- 17128287 TI - Niflumic acid inhibits chloride conductance of rat skeletal muscle by directly inhibiting the CLC-1 channel and by increasing intracellular calcium. AB - BACKGROUND AND PURPOSE: Given the crucial role of the skeletal muscle chloride conductance (gCl), supported by the voltage-gated chloride channel CLC-1, in controlling muscle excitability, the availability of ligands modulating CLC-1 are of potential medical as well as toxicological importance. Here, we focused our attention on niflumic acid (NFA), a molecule belonging to the fenamates group of non-steroidal anti-inflammatory drugs (NSAID). EXPERIMENTAL APPROACH: Rat muscle Cl(-) conductance (gCl) and heterologously expressed CLC-1 currents were evaluated by means of current-clamp (using two-microelectrodes) and patch-clamp techniques, respectively. Fura-2 fluorescence was used to determine intracellular calcium concentration, [Ca(2+)](i), in native muscle fibres. KEY RESULTS: NFA inhibited native gCl with an IC(50) of 42 muM and blocked CLC-1 by interacting with an intracellular binding site. Additionally, NFA increased basal [Ca(2+)](i) in myofibres by promoting a mitochondrial calcium efflux that was not dependent on cyclooxygenase or CLC-1. A structure-activity study revealed that the molecular conditions that mediate the two effects are different. Pretreatment with the Ca-dependent protein kinase C (PKC) inhibitor chelerythrine partially inhibited the NFA effect. Therefore, in addition to direct channel block, NFA also inhibits gCl indirectly by promoting PKC activation. CONCLUSIONS AND IMPLICATIONS: These cellular effects of NFA on skeletal muscle demonstrate that it is possible to modify CLC-1 and consequently gCl directly by interacting with channel proteins and indirectly by interfering with the calcium-dependent regulation of the channel. The effect of NFA on mitochondrial calcium stores suggests that NSAIDs, widely used drugs, could have potentially dangerous side effects. PMID- 17128288 TI - The effects induced by the sulphonylurea glibenclamide on the neonatal rat spinal cord indicate a novel mechanism to control neuronal excitability and inhibitory neurotransmission. AB - BACKGROUND AND PURPOSE: Using the neonatal rat spinal cord in vitro, we investigated the action of glibenclamide, a drug possessing dual pharmacological effects, namely block of K(ATP) channels and of the cystic fibrosis transmembrane conductance regulator (CFTR). EXPERIMENTAL APPROACH: Intra- and extracellular recordings were performed on motoneurons and interneurons. RT-PCR and western immunoblotting were used to determine gene and protein expression. KEY RESULTS: Glibenclamide (50 microM) facilitated mono- and polysynaptic reflexes, hyperpolarized motoneuron resting potential, increased action potential amplitude, decreased Renshaw cell-mediated recurrent inhibition, and increased network excitability by depressing GABA- and glycine-mediated transmission. The action of glibenclamide was mimicked by tolbutamide (500 microM) or the CFTR blocker diphenylamine-2,2-dicarboxylic acid (500 microM). The action of glibenclamide was independent from pharmacological inhibition of the Na(+)-K(+) pump with strophanthidin (4 microM) and was associated with a negative shift in the extrapolated reversal potential for CI(-) dependent synaptic inhibition. On interneurons, intracellularly-applied 8-bromo-cAMP elicited an inward current and resistance decrease; effects antagonized by the selective CFTR antagonist, CFTR(inh)-172 (5 microM). RT-PCR and western immunoblotting indicated strong expression of the CFTR in neonatal rat spinal cord. CONCLUSIONS AND IMPLICATIONS: These data suggest the CFTR expressed in motoneurons and interneurons of the neonatal spinal cord is involved in the control of Cl(-) homeostasis and neuronal excitability. CFTR appeared to contribute to the relatively depolarized equilibrium potential for synaptic inhibition, an important process to control hyperexcitability and seizure-predisposition in neonates. PMID- 17128289 TI - Use-dependent inhibition of hHCN4 by ivabradine and relationship with reduction in pacemaker activity. AB - BACKGROUND AND PURPOSE: Ivabradine, a specific and use-dependent I(f) inhibitor, exerts anti-ischaemic activity purely by reducing heart rate. The aim of this work was to characterize its effect on the predominant HCN channel isoform expressed in human sino-atrial nodes (hSAN), to determine its kinetics in HCN channels from multicellular preparations and rate-dependency of its action. EXPERIMENTAL APPROACH: RT-PCR analysis of the four HCN channel isoforms was carried out on RNAs from hSAN. Patch-clamp and intracellular recordings were obtained from CHO cells stably expressing hHCN4 and isolated SAN, respectively. Beating rate of rat isolated atria was followed using a transducer. KEY RESULTS: hHCN4 mRNAs were predominant in hSAN. Ivabradine induced a time-dependent inhibition of hHCN4 with an IC(50) of 0.5 microM. In rabbit SAN, ivabradine progressively reduced the frequency of action potentials: by 10% after 3 h at 0.1 microM, by 14% after 2 h at 0.3 microM and by 17% after 1.5 h at 1 microM. After 3h, ivabradine reduced the beating rate of rat right atria with an IC(30) of 0.2 microM. The onset of action of ivabradine was use-dependent rather than time dependent with slower effects than caesium, an extracellular I (f) blocker. Ivabradine 3 microM decreased the frequency of action potentials in SAN from guinea-pig, rabbit and pig by 33%, 21% and 15% at 40 min, respectively. CONCLUSIONS AND IMPLICATIONS: The use-dependent inhibition of hHCN4 current by ivabradine probably contributes to its slow developing effect in isolated SAN and right atria and to its increased effectiveness in species with rapid SAN activity. PMID- 17128290 TI - Structural studies of algal lectins with anti-HIV activity. AB - A number of antiviral lectins, small proteins that bind carbohydrates found on viral envelopes, are currently in pre-clinical trials as potential drugs for prevention of transmission of human immunodeficiency virus (HIV) and other enveloped viruses, such as the Ebola virus and the coronavirus responsible for severe acute respiratory syndrome (SARS). Lectins of algal origin whose antiviral properties make them candidate agents for prevention of viral transmission through topical applications include cyanovirin-N, Microcystis viridis lectin, scytovirin, and griffithsin. Although all these proteins exhibit significant antiviral activity, their structures are unrelated and their mode of binding of carbohydrates differs significantly. This review summarizes the current state of knowledge of the structures of algal lectins, their mode of binding of carbohydrates, and their potential medical applications. PMID- 17128291 TI - Fatty acid and phospholipid chlorohydrins cause cell stress and endothelial adhesion. AB - The oxidation of low-density lipoprotein (LDL) is thought to contribute to atherogenesis, which is an inflammatory disease involving activation of phagocytic cells. Myeloperoxidase, an enzyme which is able to produce hypochlorous acid (HOCl), is released from these phagocytic cells, and has been found in an active form in atherosclerotic plaques. HOCl can oxidize both the lipid and protein moiety of LDL, and HOCl-modified LDL has been found to be pro inflammatory, although it is not known which component is responsible for this effect. As HOCl can oxidize lipids to give chlorohydrins, we hypothesized that phospholipid chlorohydrins might have toxic and pro-inflammatory effects. We have formed chlorohydrins from fatty acids (oleic, linoleic and arachidonic acids) and from phospholipids (stearoyl-oleoyl phosphatidylcholine, stearoyl-linoleoyl phosphatidylcholine and stearoyl-arachidonoyl phosphatidylcholine), and investigated various biological effects of these oxidation products. Fatty acid and phospholipid chlorohydrins were found to deplete ATP levels in U937 cells in a concentration-dependent manner, with significant effects observed at concentrations of 25 microM and above. Low concentrations (25 microM) of stearoyl oleoyl phosphatidylcholine and stearoyl-arachidonoyl phosphatidylcholine chlorohydrins were also found to increase caspase-3 activity. Finally, stearoyl oleoyl phosphatidylcholine chlorohydrin increased leukocyte adhesion to artery segments isolated from C57Bl/6 mice. These results demonstrate potentially harmful effects of lipid chlorohydrins, and suggest that they may contribute to some of the pro-inflammatory effects that HOCl-modified low density lipoprotein has been found to induce. PMID- 17128292 TI - Bi-atrial subxiphoid epicardial pacemaker in superior vena cava syndrome. AB - A patient with a bi-atrial-ventricular permanent pacemaker due to paroxystic atrial fibrillation associated to sinus bradycardia, in chronic use of oral anticoagulant, presented clinical signs of superior vena cava syndrome. Digital subtraction venography showed total obstruction of the right brachiocephalic venous trunk and severe stenosis of the connection of the left trunk to the superior vena cava. The therapeutic approach consisted of complete removal of transvenous system followed by re-implant of the bi-atrial-ventricular system using an epicardial subxiphoid access with fluoroscopic assistance. PMID- 17128293 TI - Metastatic tumor of squamous cell carcinoma from uterine cervix to heart: ante mortem diagnosis. AB - Sixty-three-year-old woman with a past medical history of uterine cancer and complaint of fatigue and dyspnea on mild exertion. Physical examination revealed hypertension and rales at lung bases. A transthoracic echocardiogram showed a mass with reduced mobility in the right ventricle. The patient was taken to surgery during which a mass involving the anterior wall of the pulmonary artery, tricuspid valve, right atrium, and posterior wall of the right ventricle was found. The pulmonary artery and the right ventricle were reconstructed with a bovine pericardium patch and the tricuspid valve was replaced by a number-31 biological prosthesis. The pathological examination revealed metastasis of squamous cells with well-differentiated infiltrative areas. The patient was discharged one month after surgery. Four months later, however, she was readmitted to hospital in terminal stage, confirming the guarded prognosis of the disease at this stage. PMID- 17128294 TI - Post-transplantation lymphoproliferative disorder in pediatric patient. AB - Immunosuppressive therapy for transplanted patients exposes them to a high risk of developing posttransplantation lymphoproliferative disorders (PTLD). We report the case of a child undergoing heart transplantation at seven months of age who developed PTLD at nine years of age, diagnosed by resection of a pulmonary nodule. PMID- 17128295 TI - Acute aortic dissection during pregnancy. AB - Acute aortic dissection is one of the most dreaded clinical conditions during pregnancy. Difficulties in establishing a diagnosis and limitations regarding invasive studies increase mortality rates associated with the disease. The limited experience reported in the literature does not allow the determination of guidelines for clinical and/or surgical management of aortic dissection in pregnancy. The authors present a case of acute aortic dissection in a woman in her 33rd week of gestation and discuss the diagnostic approach considering the peculiarities of the diseases manifestation. PMID- 17128296 TI - A rare association of systemic lupus erythematosus, morbid obesity and Takotsubo Syndrome. AB - The authors report a 68-year-old white female with long-term systemic lupus erythematosus as well as morbid obesity, characterized by very elevated body mass index, who presented a classical picture of acute coronary failure ascribed to Takotsubo syndrome. PMID- 17128297 TI - Sinus node dysfunction in a patient with left atrial isomerism. AB - We report the case of an asymptomatic six-year-old child with left atrial isomerism and sinus venosus atrial septal defect. The physical examination revealed several periods of bradycardia. During a 24-hour electrocardiographic monitoring the patient presented a significant sinus node dysfunction with sinus pauses of up to 2.4 seconds. A permanent pacemaker was implanted, with a satisfactory outcome. PMID- 17128298 TI - The other side of the clinical examination in modern medicine. PMID- 17128299 TI - Fontan operation: reflections on its current evolution and perspectives. PMID- 17128300 TI - Mechanical or biological decision? PMID- 17128301 TI - Complete, incomplete or no myocardial revascularization. PMID- 17128302 TI - Surgical treatment of the ascending aorta aneurysm: simultaneous echocardiographic observation of coronary ostia reimplanted to vascular prosthesis. PMID- 17128303 TI - Cardiovascular, ventilatory, and metabolic parameters during exercise: differences between children and adults. PMID- 17128304 TI - Protein-losing enteropathy after the Fontan operation. PMID- 17128305 TI - Expert consensus (SBC/SBHCI) on the use of drug-eluting stents: recommendations of the Brazilian society of interventional cardiology/ Brazilian society of cardiology for the Brazilian public single healthcare system. AB - The authors review percutaneous coronary intervention (PCI) evolution and its growing application in myocardial revascularization for patients with coronary heart disease in Brazil and worldwide. PCI was introduced in 1977 using only the catheter balloon. Limitations of this method (acute occlusion and coronary restenosis) led to the adoption of coronary stents and more recently the advent of drug-eluting stents2, which were developed to drastically reduce restenosis rates. These developments allowed the exponential growth of percutaneous coronary intervention (PCI) procedures in Brazil which have replaced many bypass surgery procedures and have become the gold standard for the majority of symptomatic patients suffering from coronary artery disease. The preference for this procedure gained new dimensions in 2000 when the Brazilian Public Healthcare System (SUS) began reimbursing for stent procedures. This measure exemplified the importance of the Public Healthcare System's participation in incorporating medical advances and offering a high standard of cardiovascular treatment to a large portion of the Brazilian population. It is emphasized that prevention of in stent restenosis is complex due to its unpredictable and ubiquitous occurrence. Control of this condition improves quality of life and reduces the recurrence of angina pectoris, the need to perform new revascularization procedures and hospital readmissions. The overall success of the drug-eluting stents has proven to be reliable and consistent in overcoming restenosis and has some beneficial impact for all clinical and angiographic conditions. This paper discusses the adoption and criteria for the use of drug-eluting stents in other countries as well as the recommendations established by the Brazilian Society of Interventional Cardiology for their reimbursement by SUS. The incorporation of new healthcare technology involves two distinct stages. During the first stage, the product is registered with the National Health Surveillance Agency (ANVISA). During this stage the interested company submits to the regulatory agency, results from clinical studies that demonstrate the efficacy and safety of the new device or pharmaceutical product. Frequently, in addition to clinical studies, approval records for clinical use from the regulatory agencies of other countries, mainly the United States of America and the European Community are also submitted. The successful completion of this stage means that the medication or device may be prescribed or used by the physicians in Brazil. The second stage in the incorporation of new healthcare technology involves the reimbursement or financing of the treatment that was approved in the previous stage based on its efficacy and safety. This stage can be more complex than the first one since the new technology, whether a substitution for established treatment methods or the introduction of a new treatment concept, are usually more expensive. The incorporation of new technology requires a cost-effectiveness analysis so that fund administrators can make decisions based on the universal scenario of limited resources to finance healthcare with treatments that are more and more burdensome. The difficulties of funding management are aggravated by medical and social ethical implications that arise when a treatment is approved based on its efficacy and safety but is not made available to patients who could benefit greatly from it. In Brazil, assessment methods for the incorporation of new technology based on reimbursement or financing have not been fully developed for either the private healthcare plans or the Brazilian Public Healthcare System (SUS). The implementation of new technology in both healthcare systems is a slow process and frequently the implementation is a result of the requirements of patients or the organizations that represent them and at times is the result of legal proceedings or political pressure imposed by physicians and their respective scientific societies. Our objective is to review the evolution of percutaneous coronary intervention (PCI) in Brazil and its current status in view of the advent of drug-eluting stents, the growing participation of drug-eluting stents in myocardial revascularization to treat patients with coronary heart disease, as well as, to compare the regulatory standards from Brazil and other countries regarding the incorporation and recommendations for the use of this new technology. PMID- 17128306 TI - Platelet activation in distinct forms of coronary artery disease. PMID- 17128307 TI - Seasonal variation of ventricular tachycardia registered in 24-hour Holter monitoring. AB - OBJECTIVE: To evaluate the seasonal variation of ventricular arrhythmias and its correlation with ambient temperature in patients submitted to 24-hour Holter monitoring in the city of Porto Alegre, southern Brazil. METHODS: Holter monitoring reports of 3,034 patients from 1996 to 2002 were analyzed. Ventricular tachycardia (VT) was defined as the presence of 3 or more consecutive ventricular beats, at a rate equal to or higher than 100 beats per minute. Percentage distribution of patients presenting VT by seasons and its correlation with ambient temperature were analyzed. RESULTS: Mean age was 59.2 +/- 17.4 years, with a predominance of the female sex (61.9%). Patient distribution by season of the year was: 561 (18.5%) in summer, 756 (24.9%) in fall, 843 (27.8%) in winter and 874 (28.8%) in spring. Fifty-two patients (9.3%) presented VT episodes in summer, 39 (5.2%) in autumn, 56 (6.6%) in winter and 60 (6.9%) in spring (p = 0.035). There was a 40% relative increase in the proportion of patients presenting VT during summer in comparison to winter. There was a trend of increase in the proportion of patients presenting VT with rising temperatures (r = 0.57; p = 0.052). CONCLUSION: The occurrence of VT presents seasonal variations in southern Brazil, with a higher proportion of episodes occurring in summer. There is an association trend between VT and temperature increase. PMID- 17128308 TI - Criteria for central obesity in a Brazilian population: impact on metabolic syndrome. AB - OBJECTIVE: To identify and propose the optimal waist circumference cut-off points (WCp) for the diagnosis of central obesity (CO) in a Brazilian population, so as to compare these cut-off points with those recommended by the ATPIII (WC-ATPIII), and to estimate the difference in prevalence rates of metabolic syndrome (MS) using the two criteria. METHODS: Cross-sectional study conducted in a population subgroup of 1439 adults in the city of Salvador, Brazil. ROC curves of waist circumference (WC) were plotted to identify diabetes mellitus (DM) and obesity. ROC curve sensitivity and specificity values >60% and the closest to each other were used to define WCp. The prevalence of MS was estimated using WCp and WC ATPIII. RESULTS: Eight hundred and twenty nine women comprised 57.7% of the sample. The WCp selected were 84 cm for women and 88 cm for men. These cut-off points detected DM with a 68.7% and 70% sensitivity, and a 66.2% and 68.3% specificity, respectively. For obesity, sensitivity and specificity were 79.8% and 77.6% in women and 64.3% and 71.6% in men, respectively. Using WC-ATPIII, 88 cm (for women) and 102 cm (for men), the sensitivity was 53.3% and 26.5%, respectively, to diagnose DM. For obesity, sensitivity was 66.5% (for women) and 28.6% (for men). The prevalence of MS using WCp was 23.7%, 95%CI (21.6-25.9), whereas using WC-ATPIII it was 19.0%, 95%CI (17.1-20.9), 1.2 times higher using WCp. CONCLUSION: WC-ATPIII were inappropriate and underestimated the prevalence of MS in the population studied, particularly among men. We suggest that the WC cut-off points > 84 cm for women and > 88 cm for men should be tested in other Brazilian populations. PMID- 17128309 TI - Indication of myocardial perfusion scintigraphy for coronary artery disease detection based on clinical-epidemiological and treadmill test evidences. AB - OBJECTIVE: To establish when the myocardial perfusion scintigraphy (MPS) should be performed based on well-defined information obtained from treadmill test results and clinical-epidemiological parameters for coronary artery disease (CAD). METHODS: 2,100 patients who underwent MPS were classified according to the results of scintigraphy, the Duke score and a clinical-epidemiological score based on Framingham study. The patients with positive results on MPS were followed to define whether the results were true positives. Receiver operating characteristic (ROC) curves were used to establish the efficiency and the best Duke and clinical-epidemiological scores to define patients that should be submitted to scintigraphy. RESULTS: It was observed that the MPS use restriction in patients with Duke score below 7.5 and/or clinical-epidemiological score above 4 could decrease the utilization of this method by 50% without exposing the patients to a significant misdiagnosis risk. CONCLUSION: The utilization of the Duke score and a clinical-epidemiological score to classify the patients expressively decreased the number of unnecessarily requested scintigraphies. PMID- 17128310 TI - Isolated unilateral pulmonary artery agenesis: evaluation of natural and long term evolution after corrective surgery. AB - OBJECTIVE: Unilateral pulmonary artery agenesis is an uncommon isolated anomaly and since 1978 only 119 cases have been reported. In general the condition presents as pulmonary hypertension (PH) in children and hemoptysis in adults. Interventions such as pulmonary artery reconstruction and lobectomies were performed in 17% of the cases. We analyzed four of these cases, two in natural evolution and two with late term PH regression after surgical correction. METHODS: Three 22,10 and 35 month old male children and one 20 month old female child were included in the study. The 22 month old presented right-sided heart failure (RHF) and cyanosis; the 10 month old presented RHF and the other two presented exertion fatigue. All had PH symptoms, right ventricular strain on the EKG and cardiomegaly. Cardiac catheterization showed systemic pressures in the contralateral pulmonary artery, with right-sided agenesis in three of the children and left-sided agenesis in one child. RESULTS: Surgical correction of pulmonary artery continuity was possible in the 22 month old and 10 month old using a 7 mm diameter Goretex conduit between the pulmonary arteries up to the hypoplastic contralateral pulmonary hilum. There was early and late regression of the PH signs and the children remained stable during follow-up to the ages of 7 and 2.5 years, respectively. The pressure ratio between the left and right ventricles was 30 and 40%, in both cases. Pulmonary perfusion increased from 8 to 44% and from 8 to 23%, in the two cases. The same procedure was scheduled for the other patients. CONCLUSION: This technique has become the operation of choice for similar cases, that are rarely described in literature, even in the presence of severe PH and contralateral pulmonary artery hypoplasia. PMID- 17128311 TI - Independent predictors of in-hospital outcomes following coronary stent implantation. AB - OBJECTIVE: To identify the clinical and angiographic predictors of early outcome following coronary stent implantation. METHODS: Nine hundred and forty-six patients (pt) [61.04 +/- 10.98 years old, range 31 to 91] underwent stent implantation; 580 male (61.3%). Procedural success was defined when a pt had at least one vessel successfully dilated with a residual stenosis < 20%. Clinical success occurred when a pt had at least one vessel successfully dilated without a major complication (MC) [death, myocardial infarction (MI), coronary artery bypass graft] during the hospital stay. Clinical and angiographic determinants of outcome were studied. All variables related to early outcome evaluated by univariate analysis were included in a multiple logistic regression analysis (MLR). RESULTS: Procedural success was achieved in 98.9%; clinical success in 95.7%; an unsuccessful uncomplicated outcome in 0.1% and major complications in 4.2%. By MLR, procedural success was related to restenotic lesion, calcification, and irregular contour. Clinical success was related to diabetes mellitus, cardiogenic shock, acute coronary syndromes, age, left ventricular dysfunction, calcification, and total occlusion. Major complications was predicted by diabetes mellitus, cardiogenic shock, acute coronary syndromes, age, multivessel disease, left ventricular dysfunction, calcification, long lesions, and total occlusions. Mortality was predicted by cardiogenic shock, acute coronary syndromes, age, arterial hypertension, and left ventricular dysfunction. CONCLUSION: These results suggest that early outcome was significantly affected by cardiogenic shock, left ventricular dysfunction, age, calcification, and total occlusion. PMID- 17128312 TI - The influence of time on the characteristics and risk factors for patients submitted to myocardial revascularization. AB - OBJECTIVE: To compare the clinical and surgical profile between two groups of patients submitted to Myocardial Revascularization (MCR) surgery at the Instituto de Cardiologia of Rio Grande do Sul with a ten year interval, to observe its influence upon MCR hospital mortality and to verify the predictability of this result using the risk score. METHODS: A retrospective cohort study involving 307 patients who underwent MCR surgery within a six month period during 1991/92 (INITIAL group, n=153) or 2001/02 (CURRENT group, n=154). Demographic characteristics, heart disease, comorbidities and surgical events were analyzed to compare the groups and to define the hospital mortality risk score (based on the Cleveland Clinic method). RESULTS: The CURRENT group was older, had more severe heart condition (functional class, incidence of heart failure and number of vessels with severe lesions) and a greater prevalence of comorbidities. The INITIAL group had a higher prevalence of nonelective surgery. Both groups had similar mean risk scores (2.8 +/- 3.1 for INITIAL and 2.2 +/- 2.5 for CURRENT) and hospital mortality rates (3.3% and 1.9% respectively). These figures are comparable to those for reported by Cleveland Clinic (for a risk score of 3 the predicted mortality range between 2.0 %; using a confidence level of 95% the predicted mortality is between 0 and 4.3%; and actual mortality confirmed by the study was 3.4%). CONCLUSION: Patients currently submitted to MCR are older and in worse clinical condition (heart and systemic) than those operated on ten years ago; however, the risk scores and hospital mortality rates were slightly higher in the INITIAL group. The higher number of nonelective surgical interventions could have contributed to this. A risk score can be used to identify patients that require a higher level of care and to predict surgical outcomes. PMID- 17128313 TI - Platelet activation in different clinical forms of the coronary artery disease (role of P-selectin and others platelet markers in stable and unstable angina). AB - OBJECTIVE: Markers of platelet activation are elevated in coronary artery disease. We sought to identify the presence and the potential associations of different markers of platelet activation. METHODS: We studied patients with unstable angina (n=28), patients with stable angina (n=36) and patients without coronary artery disease (n=30); sex and age matched. Blood levels of the adhesion molecule P-selectin, Thromboxane B2 and Serotonin were measured by enzyme immunoassays. RESULTS: When we compared the groups the results were: sP-selectin, thromboxane B2 and serotonin levels were significantly higher in patients with unstable angina than in patients with stable angina. CONCLUSION: These markers of platelet activation were able to identify unstable forms of coronary artery disease. PMID- 17128314 TI - Functional assessment of coronary grafts on dobutamine pharmacological stress echocardiogram. AB - OBJECTIVE: To verify the sensitivity, specificity and diagnostic accuracy of dobutamine stress echocardiogram (DSE) when assessing the functional status of coronary grafts: sufficient (SUF) or insufficient (INS). METHODS: We carried out a prospective, observational study which included 25 patients submitted to coronary artery bypass grafting (CABG). The DSE and the coronary angiography were performed before the CABG and three months after the CABG. The left ventricle was divided into three territories per patient according to the three major coronary arterie: the anterior descending (AD), the circumflex (CX) and the right coronary (RC). Of the 75 possible territories, 54 were revascularized: 25 were specific to the AD artery and 29 of the CX/RC arteries. INS means luminal obstruction or occlusion greater than or equal to 50%. RESULTS: In 14 (26%) of the 54 revascularized territories the grafts were INS. The DSE detected ischemia in 16 (28%) territories; 10 of which had INS grafts. The DSE detected ischemia in 6 (15%) of the 40 territories whose grafts were SUF. Therefore, the DSE had a sensitivity of 71.4%, specificity of 85% and diagnostic accuracy of 81.4%. CONCLUSION: The DSE is a diagnostic method with high specificity and diagnostic accuracy, and good sensitivity for the functional assessment of coronary grafts. PMID- 17128315 TI - Prognostic value of the Doppler index of myocardial performance in postoperative of coronary artery bypass surgery. AB - OBJECTIVE: Myocardial Performance Index (MPI) obtained by Doppler echocardiography for the non-geometrical evaluation of systolic and diastolic function has been described as a method for prognostic evaluation in patients with acute myocardial infarction (AMI). Using the same condition, the objective of this study was to evaluate the predictive value of MPI for cardiovascular complications in patients at low risk during the postoperative period of CABG. METHODS: Eighty patients submitted to CABG with adequate left ventricular function in the preoperative period were studied, with MPI measured during the first hours postoperatively. Patients were followed until hospital discharge. Statistical analysis included Chi-Square test, Student t test, Mann-Whitney test, and estimation of relative risks with 95% confidence intervals, sensitivity and specificity plus a ROC curve. RESULTS: The data were evaluated by two independent observers blinded to the clinical data with non-significant intra and interobserver variability. MPI=0.43 was found as the cutoff point, considering patients with a higher probability of postoperative events those who had MPI above 0.43. The relevant events for analysis were AMI (RR 0.87 ci 0.21-3.65), atrial fibrillation (RR 0.65 ci 0.24 - 1.76), other arrhythmias (RR 1.51 ci 0.36 6.33), LV dysfunction (RR 1.74 ci 0.32-9.88), with no association between patients with MPI>0.43 and the occurrence of these events. CONCLUSION: No association was found between MPI and cardiovascular complications and longer hospital stay in this group of patients, and this index was considered not adequate as an isolated predictive method. PMID- 17128316 TI - Waist circumference and body mass index as predictors of hypertension. AB - OBJECTIVE: To evaluate the association between anthropometric indexes--body mass index (BMI) and waist circumference (WC)--and hypertension, and to evaluate the predictive value of these indexes in detecting hypertension. METHODS: Cross sectional population study conducted in the city of Goiania (GO) with a sample of 1,238 adults aged twenty to 64 years, in 2001. Total obesity was defined as BMI > or = 30 kg/m(2); abdominal obesity was defined as level 2 WC > or = 88 cm for women and > or = 102 cm for men, and hypertension was defined as systolic pressure > or = 140 mmHg, or diastolic pressure > or = 90 mmHg, or utilization of hypotensive drugs). Multiple logistic regression analysis was used to evaluate the associations between anthropometric indexes and hypertension. The Receiver Operating Characteristic (ROC) curve analysis was used to evaluate sensitivity and specificity of BMI (> or = 30) and level 2 WC as predictive factors of hypertension, and to determine the best predictive cut-off points for hypertension. RESULTS: WC was associated with hypertension in both genders. Level 2 WC and BMI > or = 30 kg/m(2) showed a low sensitivity in identifying hypertension. The best predictive cut-off points for hypertension coincided with level 1 WC (> or = 80 cm) and with BMI > or = 25 kg/m(2) (overweight) for women, and were lower than the values of level 1 WC and of overweight for men. CONCLUSION: Level 2 WC and BMI > or = 30 kg/m(2) are not adequate to identify the groups at the highest risk of hypertension, since this risk rises with small increases in adiposity. PMID- 17128317 TI - Risk factors for hypertension and diabetes mellitus in metallurgic and siderurgic company's workers. AB - OBJECTIVE: To study the factors associated with arterial hypertension (AH) and diabetes mellitus (DM) in a metal and steel industry's workers with units in Sao Paulo and Rio de Janeiro, using a hierarchical risk model. METHODS: This is a cross-sectional study. We obtained information on demographic, occupational and lifestyle variables, in addition to the lipid profile, fasting glycemia and blood pressure of 3,777 employees, and we carried out descriptive statistical analysis and hierarchical multiple logistic regression analysis. RESULTS: The prevalence of AH was 24.7% and the hierarchical regression analysis indicated that male gender and age above 40 years presented statistically significant risk. Regardless of demographic characteristics, working in the metal industry, intense stress at work, sedentary lifestyle, alcohol consumption, body mass index above 25, altered cholesterol and altered triglycerides were associated with AH. The prevalence of DM was 11.5% and the hierarchical regression analysis indicated that male gender and age above 40 years presented statistically significant risk. Regardless of demographic characteristics, the same conditions were associated with DM. CONCLUSION: The data evidenced that workers above 40 years are a priority for intervention actions aimed at improving the prevention of these two conditions. These actions should focus especially on eating habits and the practice of physical exercise, which could improve the control of obesity and of alterations in the lipid profile. PMID- 17128318 TI - An evaluation of the Rastreometro, a new device for populational screening for high blood pressure in developing countries. AB - OBJECTIVE: To test a simplified blood pressure device called Rastreometro that could be used by the Health Agents. METHODS: The Rastreometro has been developed from an ordinary aneroid sphygmomanometer, in which the numeric display is covered by an adhesive with a red zone, indicating pressures equal or above 140 mmHg and a yellow zone indicating pressures below 140 mmHg. The onset of oscillations of the aneroid needle is taken as an indication of the systolic pressure value. The measurements made by the Rastreometro were compared with those made by the auscultatory method, and were carried out in 268 patients, by two operators. The influence on the results of confounding variables such as age, gender, BMI, arm length, upper arm circumference, skin colour and antihypertensive treatment were taken into consideration, as well as intra and interobserver variation. RESULTS: In the whole group, sensitivity was 95.1%, specificity was 63.1%, positive predictive value was 62.4% and negative predictive value was 95.3%. Hypertensive treatment significantly affected specificity, 32.7% as compared to 77.8% for the non-treated group. Both operators improved their results over time. CONCLUSION: This study suggests that the Rastreometro technique, as a screener for hypertension, has good sensitivity. Concerning specificity, it is acceptable, provided the patient is not on regular antihypertensive treatment. In this latter situation, it can be improved by a proper standardization of the method to read the systolic pressure by needle oscillations. Furthermore, the use of this technique requires well trained operators. PMID- 17128319 TI - Predictive factors for pregnancy hypertension in primiparous adolescents: analysis of prenatal care, ABPM and microalbuminuria. AB - OBJECTIVE: To quantify PH prevalence in primiparous adolescents; define predictive factors for the occurrence of PH and its impact on newborns. METHODS: We followed 29 primiparous adolescents from the prenatal period through the 12th week of the puerperium, with a mean of sixteen years of age, served at the Outpatient Facility for Adolescents of Maternidade Escola Assis Chateaubriand (MEAC) of Universidade Federal do Ceara (Fortaleza, Brazil). The pregnant adolescents were divided into two groups, that is, those who remained normotensive (Group I) and those who developed PH (Group II). The variables investigated in the assessment of the value of predictability for the development of PH were anthropometric measures, socioeconomic aspects, smoking habit, inheritance for SAH (father/mother), prenatal tests requested in the first prenatal care visit in addition to microalbuminuria and ambulatory blood pressure monitoring (ABPM) in the 28th week of gestation. The pregnant adolescents were followed up at delivery and late puerperium (12th week after the puerperium). The newborns to the mothers included in our study were assessed at birth according to the Apgar score and the Capurro method, for weight, height and perinatal hypoxia. RESULTS: The prevalence of PH was 51.7%. Inheritance for SAH presented the highest predictive value for PH with an odds ratio of 10.99. Diastolic arterial pressure equal to or above 70 mmHg at the gestational age of 35 weeks was statistically significant as a predictive value for PH. At ABPM we found a predictive value for PH: diastolic pressure load during alertness, diastolic and systolic pressure load during night sleep, pressure variability and maximum diastolic pressure during sleep. Specifically a maximum diastolic arterial pressure (DAP) at ABPM during the period of night sleep (3)64 mmHg presented an odds ratio of 6 for PH with a sensitivity of 80% and a specificity of 60% for the development of PH. CONCLUSION: The research for PH predictive factors in primiparous adolescents showed to be easy to apply and useful to stratify high risk pregnant women as regards the development of PH. PMID- 17128320 TI - Arterial hypertension and family care: care provided to hypertensive individuals in a small municipality in Southern Brazil. AB - OBJECTIVE: Based on the analysis of indicators which express the rationale of the actions of the program, we developed a cross-sectional study to assess the performance of a Family Healthcare Team as regards the care provided to individuals with arterial hypertension living in a small municipality. METHODS: The study carried out was based on the analysis of the records (Record A) of SIAB (Primary Care Information System) of 418 individuals suffering from arterial hypertension, of 351 records stored in the primary healthcare center, and of 376 interviews with individuals living in the area under the team's care, in the year 2003. The following variables were studied: gender, age, family income, housing conditions, health services used, presence of diabetes mellitus, coverage and concentration of care. RESULTS: Women above 60 years of age prevailed. They were 65.7% of the cases. The arterial hypertension prevalence rate was 18%, and was different for both genders: for women it was 22.3%. The coverage was 64.1% for appointments with doctors, 32.4% for appointments with nurses, and 36.4% for home visits. The information on files revealed that 25.8% of hypertensive individuals did not receive care in 2003 and 52.7% did not receive care in the last semester. The concentration of care was below the recommendation. CONCLUSION: The information showed that the organization of health care does not reflect the rationale of health surveillance in that services focus mostly on meeting spontaneous demand, which in turn reflects unpreparedness as to the use of information systems for the planning and evaluation of actions. PMID- 17128321 TI - Twenty-four hour blood pressure record for smokers and nonsmokers. AB - OBJECTIVE: To evaluate the effect of smoking on blood pressure trends during a 24 hour period, by analyzing the parameters of the ambulatory blood pressure monitoring (ABPM). METHODS: The results of 289 ABPM tests conducted on patients classified as smokers or nonsmokers were studied. The parameters analyzed were: mean 24-hour, daytime and nighttime systolic and diastolic blood pressure readings; nocturnal dipping of systolic and diastolic pressures and blood pressure load. The patients were classified in four groups according to whether or not they used antihypertensive medication: 1A--nonsmokers using medication; 1B -smokers using medication; 2A--nonsmokers not using medication; and 2B--smokers not using medication. Variables were expressed as minimum, maximum, median, mean and standard deviation values. Univariate analysis was used for comparing the smoking and nonsmoking groups. The significantly different variables from the groups were selected using multivariate analysis. The significance level adopted was 5%. RESULTS: Mean daytime systolic and diastolic blood pressures were significantly higher in the smokers, regardless of whether or not they used antihypertensive medication. Mean nocturnal blood pressure readings were similar between smokers and nonsmokers. Mean 24-hour systolic blood pressure readings were significantly higher in the smokers, regardless of whether or not they used antihypertensive medication. Nocturnal dipping was similar for all groups. Blood pressure loads were consistently and significantly higher in the smokers regardless of medication use. CONCLUSION: Mean daytime systolic and diastolic blood pressure readings were consistently higher in the smokers when compared to nonsmokers regardless of antihypertension medication use. Nocturnal dipping was similar for smokers and nonsmokers. PMID- 17128322 TI - Vascular radiolesion as a deleterious effect of high-dose-rate intraarterial brachytherapy with samarium-153 in hypercholesterolemic rabbits. AB - OBJECTIVE: This study was designed to evaluate vascular morphological and morphometric changes induced by brachytherapy with samarium-153 (Sm-153) at high doses in hypercholesterolemic rabbits. METHODS: Forty-three New Zealand White hypercholesterolemic rabbits were analyzed, and the total of 86 iliac arteries underwent balloon angioplasty injury. The rabbits were divided into three different groups: two irradiation groups (IG) assigned to 15 Gy (n=14) and 60 Gy (n=36) irradiation doses, respectively, and a control group (n = 36). Histomorphometric and qualitative histological analyses were performed for tissue evaluation. RESULTS: Significant reductions were found in neointimal proliferation (NIP) (p< 0.0001), media area (MA) (p<0.0001) and percent stenosis (p<0.0001) in the 15-Gy IG, compared to the other groups. The 60-Gy IG had the higher rate of NIP, increase in media and vessel areas (VA) and percent stenosis. The 60-Gy IG also showed the greatest number of xanthomatous cells (60-Gy IG: 86.11% and 15-Gy IG: 14.29%, p<0.0001) and the highest amount of hyaline amorphous tissue (60-Gy IG:58.33% and 15-Gy IG:0%, p=0.0001) and vascular proliferation (60-Gy IG:30.56% and 15-Gy IG:0%, p=0.0221). No statistically significant differences were found among groups concerning other tissue analyses. CONCLUSION: The high-dose irradiation of 60 Gy resulted in intense cell proliferation considered vascular radiolesion, unlike the 15-Gy dose, which was associated with an excellent inhibition of neointimal proliferation. PMID- 17128323 TI - The effect of L-arginine on neointimal proliferation and artery remodeling on an iliac artery lesion caused by a balloon catheter in hypercholesterolemic rabbits. AB - OBJECTIVE: It has been implied that neointimal proliferation and remodeling are the major causes of restenosis. The objective of this study is to assess the effect of orally administered L-arginine on these two factors in hypercholesterolemic rabbits that had suffered an injury to their iliac arteries caused by a catheter balloon. METHODS: The study included nineteen rabbits that were divided in two groups: control (CG) and arginine (AG). There were 19 arteries studied from the control group and 17 in the arginine group. The animals were placed on a 2% hypercholesterolemic diet for 15 days and then submitted to a balloon angioplasty in order to produce a lesion in their iliac arteries. Next, the AG animals were given a 1g/kg/day oral dose of a L-arginine solution. The animals were sacrificed 15 days after the angioplasty procedure and histological artery sections were prepared, stained and fixed. The ratio between the neointimal area (in mm(2)) and the media layer (in mm(2)) was used to represent lesion development. In order to determine remodeling, the ratio between the total area of the medial portion of the vessel (greater balloon contact) and the total area of the reference segment of the vessel (less balloon contact) was used. RESULTS: Mean neointimal thickness (NI/M) was 0.8151+/-0.2201 in CG and 0.3296+/ 0.1133 in AG. Remodeling patterns for the two groups studied were similar. CONCLUSION: In the experimental model used, L-arginine was able to reduce intimal tissue thickness in hypercholesterolemic rabbits but did not act on artery remodeling. PMID- 17128324 TI - Hemodynamic and vascular endothelium function studies in healthy pigs after intravenous bolus infusion of methylene blue. AB - OBJECTIVE: Clinical benefit of methylene blue (MB) treating NO-induced vasoplegia has been reported in sepsis, systemic inflammatory response syndrome (SIRS) in cardiac surgery and anaphylactic shock, but its safety is sometimes questioned, mainly regarding its hemodynamic effects and the possibility of causing endothelium dysfunction. To examine the nitric oxide plasma levels and cardiovascular effects of the infusion of MB in vivo and its effects on endothelium-dependent and endothelium-independent in vitro vascular relaxation. METHODS: The study protocol included two experimental groups of female pigs: Group I (Control) - the animals (n=6) did not receive MB; Group II (MB)--the animals received 3 mg/kg of MB intravenous bolus infusion. After fifteen minutes of hemodynamic parameter recording the animals were sacrificed by exsanguination, and in vitro studies were conducted using segments of coronary, hepatic, superior mesenteric and renal arteries, to determine the effect of MB on the arterial endothelium function with regard to NO release. Nitric oxide plasma levels (NOx) were measured in each of the experimental groups. RESULTS: The results obtained in the present investigation were: 1) intravenous infusion of MB (3.0 mg/kg) caused no hemodynamic changes; 2) absolute and percent plasma NOx values did not differ between the experimental groups; and 3) in vitro study of vascular relaxation showed no significant difference between groups. These results show that MB intravenous infusion seems to be safe. This finding agrees with data from clinical experiments where MB was used to treat vasoplegic syndrome after cardiopulmonary bypass, systemic inflammatory response syndrome (SIRS) and anaphylaxis. These results were not unexpected because, as in healthy subjects, hemodynamics is only fine tuned and not fully under NO control; therefore, MB inhibiting guanylyl cyclase is not expected to do anything. CONCLUSION: Intravenous use of MB, at the investigated dose, did not cause any abnormal hemodynamic responses or impairment of endothelium-dependent relaxation. PMID- 17128325 TI - Case 3/2006: nineteen-month-old infant with pulmonary atresia and ventricular septal defect and decreased pulmonary blood flow. PMID- 17128326 TI - Magnetic resonance imaging is image diagnosis in heart valve disease. PMID- 17128327 TI - Autonomic dysregulation and the metabolic syndrome: pathologic partners in an emerging global pandemic. PMID- 17128328 TI - The endocannabinoid system: a new perspective for cardiometabolic risk control. PMID- 17128329 TI - Are drugs for rare diseases "essential"? PMID- 17128330 TI - Health and foreign policy. PMID- 17128331 TI - Reaching the targets for TB control: call for papers. PMID- 17128332 TI - WHO launches taskforce to fight counterfeit drugs. PMID- 17128333 TI - Free access to journals gives Kenyan science a boost. PMID- 17128334 TI - Can better health statistics save lives? PMID- 17128336 TI - A century in public health. PMID- 17128337 TI - WHO response to the humanitarian crisis in Lebanon. PMID- 17128339 TI - Causes of stillbirths and early neonatal deaths: data from 7993 pregnancies in six developing countries. AB - OBJECTIVE: To report stillbirth and early neonatal mortality and to quantify the relative importance of different primary obstetric causes of perinatal mortality in 171 perinatal deaths from 7993 pregnancies that ended after 28 weeks in nulliparous women. METHODS: A review of all stillbirths and early newborn deaths reported in the WHO calcium supplementation trial for the prevention of pre eclampsia conducted at seven WHO collaborating centres in Argentina, Egypt, India, Peru, South Africa and Viet Nam. We used the Baird-Pattinson system to assign primary obstetric causes of death and classified causes of early neonatal death using the International classification of diseases and related health problems, Tenth revision (ICD-10). FINDINGS: Stillbirth rate was 12.5 per 1000 births and early neonatal mortality rate was 9.0 per 1000 live births. Spontaneous preterm delivery and hypertensive disorders were the most common obstetric events leading to perinatal deaths (28.7% and 23.6%, respectively). Prematurity was the main cause of early neonatal deaths (62%). CONCLUSIONS: Advancements in the care of premature infants and prevention of spontaneous preterm labour and hypertensive disorders of pregnancy could lead to a substantial decrease in perinatal mortality in hospital settings in developing countries. PMID- 17128340 TI - Rates, timing and causes of neonatal deaths in rural India: implications for neonatal health programmes. AB - OBJECTIVE: To assess the rates, timing and causes of neonatal deaths and the burden of stillbirths in rural Uttar Pradesh, India. We discuss the implications of our findings for neonatal interventions. METHODS: We used verbal autopsy interviews to investigate 1048 neonatal deaths and stillbirths. FINDINGS: There were 430 stillbirths reported, comprising 41% of all deaths in the sample. Of the 618 live births, 32% deaths were on the day of birth, 50% occurred during the first 3 days of life and 71% were during the first week. The primary causes of death on the first day of life (i.e. day 0) were birth asphyxia or injury (31%) and preterm birth (26%). During days 1-6, the most frequent causes of death were preterm birth (30%) and sepsis or pneumonia (25%). Half of all deaths caused by sepsis or pneumonia occurred during the first week of life. The proportion of deaths attributed to sepsis or pneumonia increased to 45% and 36% during days 7 13 and 14-27, respectively. CONCLUSION: Stillbirths and deaths on the day of birth represent a large proportion of perinatal and neonatal deaths, highlighting an urgent need to improve coverage with skilled birth attendants and to ensure access to emergency obstetric care. Health interventions to improve essential neonatal care and care-seeking behavior are also needed, particularly for preterm neonates in the early postnatal period. PMID- 17128341 TI - Community surveys and risk factor analysis of human alveolar and cystic echinococcosis in Ningxia Hui Autonomous Region, China. AB - OBJECTIVE: To determine the true community prevalence of human cystic (CE) and alveolar (AE) echinococcosis (hydatid disease) in a highly endemic region in Ningxia Hui, China, by detecting asymptomatic cases. METHODS: Using hospital records and "AE-risk" landscape patterns we selected study communities predicted to be at risk of human echinococcosis in Guyuan, Longde and Xiji counties. We conducted community surveys of 4773 individuals from 26 villages in 2002 and 2003 using questionnaire analysis, ultrasound examination and serology. FINDINGS: Ultrasound and serology showed a range of prevalences for AE (0-8.1%; mean 2%) and CE (0-7.4%; mean 1.6%), with the highest prevalence in Xiji (2% for CE, 2.5% for AE). There were significant differences in the prevalence of CE, AE and total echinococcosis between the three counties and villages (with multiple degrees of freedom). While hospital records showed 96% of echinococcosis cases attributable to CE, our survey showed a higher prevalence of human AE (56%) compared to CE (44%). Questionnaire analysis revealed that key risk factors for infection were age and dog ownership for both CE and AE, and Hui ethnicity and being female for AE. Drinking well-water decreased the risk for both AE and CE. CONCLUSION: Echinococcosis continues to be a severe public health problem in this part of China because of unhygienic practices/habits and poor knowledge among the communities regarding this disease. PMID- 17128342 TI - Is mortality from heart failure increasing in Australia? An analysis of official data on mortality for 1997-2003. AB - OBJECTIVE: To assess whether trends in mortality from heart failure (HF) in Australia are due to a change in awareness of the condition or real changes in its epidemiology. METHODS: We carried out a retrospective analysis of official data on national mortality data between 1997 and 2003. A death was attributed to HF if the death certificate mentioned HF as either the underlying cause of death (UCD) or among the contributory factors. FINDINGS: From a total of 907 242 deaths, heart failure was coded as the UCD for 29 341 (3.2%) and was mentioned anywhere on the death certificate in 135 268 (14.9%). Between 1997 and 2003, there were decreases in the absolute numbers of deaths and in the age-specific and age-standardized mortality rates for HF either as UCD or mentioned anywhere for both sexes. HF was mentioned for 24.6% and 17.8% of deaths attributed to ischaemic heart disease and circulatory disease, respectively, and these proportions remained unchanged over the period of study. In addition, HF as UCD accounted for 8.3% of deaths attributed to circulatory disease and this did not change materially from 1997 to 2003. CONCLUSION: The decline in mortality from HF measured as either number of deaths or rate probably reflects a real change in the epidemiology of HF. Population-based studies are required to determine accurately the contributions of changes in incidence, survival and demographic factors to the evolving epidemiology of HF. PMID- 17128343 TI - The syndromic management of vaginal discharge using single-dose treatments: a randomized controlled trial in West Africa. AB - OBJECTIVE: To evaluate whether single-dose treatments are as effective as standard therapy in the syndromic management of vaginal discharge. METHODS: A randomized controlled effectiveness trial compared single-dose tinidazole plus fluconazole (TF) with treatment for 7 days with metronidazole plus 3 days of treatment with vaginal clotrimazole (MC) among 1570 women presenting with vaginal discharge at primary health care institutions in Ghana, Guinea, Mali and Togo. Participants were randomly allocated to one of the two treatments by research nurses or physicians using precoded envelopes. Effectiveness was assessed by symptomatic response on day 14. CLINICAL IDENTIFIER ClinicalTrials.gov NCT00313131. FINDINGS: The two treatment regimens had similar effectiveness: complete resolution was seen in 66% (TF) and 64% (MC) and partial resolution in 33% (TF) and 34% (MC) of participants (P = 0.26). Effectiveness was similar among subgroups with vulvovaginal candidiasis, Trichomonas vaginalis vaginitis or bacterial vaginosis. The two treatment regimens had a similar effectiveness among human immunodeficiency virus (HIV)-infected (TF: n = 76, 71% complete resolution, 28% partial; MC: n = 83, 72% complete resolution, 25% partial, P = 0.76) and HIV uninfected women (TF: n = 517, 68% complete, 32% partial; MC: n = 466, 65% complete, 33% partial, P = 0.20). Cervical infections with Neisseria gonorrhoeae, Chlamydia trachomatis and Mycoplasma genitalium were uncommon among women not involved in sex work, were associated with bacterial vaginosis or T. vaginalis vaginitis, and did not alter response to treatment with agents active against vaginal infections. Four-fifths of women not relieved by a single dose of TF had a favourable response when MC was administered as second-line treatment. CONCLUSION: Single-dose TF is as effective as multiple-dose MC in the syndromic management of vaginal discharge, even among women with HIV-infection. Given its low price and easier adherence, TF should be considered as a first-line treatment for vaginal discharge syndrome. PMID- 17128344 TI - Measuring the impact of intimate partner violence on the health of women in Victoria, Australia. AB - OBJECTIVE: Using burden of disease methodology, estimate the health risks of intimate partner violence (IPV) among women in Victoria, Australia. METHODS: We calculated population attributable fractions (from survey data on the prevalence of IPV and the relative risks of associated health problems in Australia) and determined health outcomes by applying them to disability-adjusted life year estimates for the relevant disease and injury categories for Victoria, Australia for 2001. FINDINGS: For women of all ages IPV accounted for 2.9% (95% uncertainty interval 2.4-3.4%) of the total disease and injury burden. Among women 18-44 years of age, IPV was associated with 7.9% (95% uncertainty interval 6.4-9.5%) of the overall disease burden and was a larger risk to health than risk factors traditionally included in burden of disease studies, such as raised blood pressure, tobacco use and increased body weight. Poor mental health contributed 73% and substance abuse 22% to the disease burden attributed to IPV. CONCLUSION: Our findings suggest that IPV constitutes a significant risk to women's health. Mental health policy-makers and health workers treating common mental health problems need to be aware that IPV is an important risk factor. Future research should concentrate on evaluating effective interventions to prevent women being exposed to violence, and identifying the most appropriate mental health care for victims to reduce short- and long-term disability. PMID- 17128345 TI - Rare essentials: drugs for rare diseases as essential medicines. AB - Since 1977, the WHO Model List of Essential Medicines (EML), published by WHO, has provided advice for Member States that struggle to decide which pharmaceutical technologies should be provided to patients within their public health systems. Originating from outside WHO, an incentive system has been put in place by various governments for the development of medicines for rare diseases ("orphan drugs"). With progress in pharmaceutical research (e.g. drugs targeted for narrower indications), these medicines will feature more often on future public health agendas. However, when current definitions for selecting essential medicines are applied strictly, orphan drugs cannot be part of the WHO Essential Medicines Programme, creating the risk that WHO may lose touch with this field. In our opinion WHO should explicitly include orphan drugs in its policy sphere by composing a complementary Orphan Medicines Model List as an addition to the EML. This complementary list of "rare essentials" could aid policy-makers and patients in, for example, emerging countries to improve access to these drugs and stimulate relevant policies. Furthermore, inconsistencies in the current EML with regard to medicines for rare diseases can be resolved. In this paper we propose selection criteria for an Orphan Medicines Model List that could form a departure point for future work towards an extensive WHO Orphan Medicines Programme. PMID- 17128346 TI - Can public-private collaboration promote tuberculosis case detection among the poor and vulnerable? AB - Private-public mix (PPM) DOTS is widely advocated as a DOTS adaptation for promoting progress towards the international tuberculosis (TB) control targets of detecting 70% of TB cases and successfully treating 85% of these. Private health care plays a central role in health-care provision in many developing countries that have a high burden of TB. It is therefore encouraging that PPM projects are being set up in various countries around the world to explore possible interaction between the national TB programmes and other partners in the fight against TB. The objective of this review was to use the published literature to assess the range of providers included in PPMs for their ability to provide case detection services for the vulnerable. From a case-detection perspective, we identify the essential elements of a pro-poor PPM model, namely, cost effectiveness from a patient perspective, accessibility, acceptability and quality. The review revealed that a very large part of the total spectrum of potential PPM-participating partners has not yet been explored; current models focus on private-for-profit health-care providers and non-governmental organizations. We conclude that it is important to think critically about the type of private providers who are best suited to meeting the needs of the poor, and that more should be done to document the socioeconomic status of patients accessing services through PPM pilots. PMID- 17128347 TI - One in a million: the first community trial of water fluoridation. PMID- 17128348 TI - Ethical approval for operational research. PMID- 17128349 TI - Preparing for the next natural disaster--need for a WHO coordinating centre. PMID- 17128350 TI - IMCI: what can we learn from an innovation that didn't reach the poor? PMID- 17128351 TI - Openness is key in fight against disease outbreaks. PMID- 17128353 TI - Treating malaria at home in Uganda. PMID- 17128354 TI - Street food boom in Ghana spurs calls for better hygiene. PMID- 17128355 TI - D for diagnosis and E for everything else. PMID- 17128357 TI - Treatment outcomes and risk factors for relapse in patients with early-stage human African trypanosomiasis (HAT) in the Republic of the Congo. AB - OBJECTIVE: In 2002-03, the Republic of the Congo increased the threshold separating stage 1 and 2 cases of human African trypanosomiasis (HAT) from a cerebrospinal fluid (CSF) white cell count of 5 cells/mm(3) to 10 cells/mm(3). We aimed to assess whether the increased threshold of 10 cells/mm(3) is a safe indicator of stage 2 disease. METHODS: We assessed patients treated for stage 1 HAT caused by Trypanosoma brucei gambiense in the Republic of the Congo between April 2001 and April 2005. Patients with 0-10 cells/mm(3) in CSF were classed as stage 1 and treated with pentamidine. Patients with CSF of > 10 cells/mm(3) were classed as stage 2 and treated with either melarsoprol or eflornithine. We did a retrospective analysis of all patients treated after the September 2002 increase in threshold for classification of HAT disease stage 2, and who were eligible for at least 1 year of follow-up. Primary outcome was survival without death or relapse within 1 year of discharge. Risk factors for treatment failure, in particular CSF white cell count on diagnosis, were assessed. FINDINGS: Between September 2002 to April 2004, 692 patients eligible for our analysis were treated with pentamidine. All were discharged alive. Relapse rate was 5% (n = 33). The only identified risk factor for relapse was a CSF white cell count of 6-10 cells/mm(3) rather than 0-5 cells/mm(3) (adjusted hazard ratio 3.27 (95% confidence interval, 1.52-7.01); P = 0.002). CONCLUSION: A threshold of 5 white cells/mm(3) in CSF is safer than 10 cells/mm(3) to determine stage 2 HAT and reduce risk of relapse. PMID- 17128358 TI - Melarsoprol versus eflornithine for treating late-stage Gambian trypanosomiasis in the Republic of the Congo. AB - OBJECTIVE: To compare the effectiveness of melarsoprol and eflornithine in treating late-stage Gambian trypanosomiasis in the Republic of the Congo. METHODS: We analysed the outcomes of death during treatment and relapse within 1 year of discharge for 288 patients treated with eflornithine, 311 patients treated with the standard melarsoprol regimen and 62 patients treated with a short-course (10-day) melarsoprol regimen between April 2001 and April 2005. FINDINGS: A total of 1.7% (5/288) of patients treated with eflornithine died compared with 4.8% (15/311) of those treated with standard melarsoprol and 6.5% (4/62) of those treated with short-course melarsoprol. Patients treated with eflornithine tended to be younger and were more likely to have trypanosomes or higher white blood cell counts in their cerebrospinal fluid. The cumulated incidence of relapse among patients who attended at least one follow-up visit 1 year after discharge was 8.1% (11/136) for those treated with eflornithine, 14% (36/258) for those treated with standard melarsoprol and 15.5% (9/58) for those treated with shortcourse melarsoprol. In a multivariate analysis, when compared with eflornithine, standard melarsoprol was found to be a risk factor for both death (odds ratio (OR) = 2.87; 95% confidence interval (CI) = 1.03-8.00) and relapse (hazard ratio (HR) = 2.47; 95% CI = 1.22-5.03); when compared with eflornithine, short-course melarsoprol was also found to be a risk factor for death (OR = 3.90; 95% CI = 1.02-14.98) and relapse (HR = 6.65; 95% CI = 2.61 16.94). CONCLUSION: The effectiveness of melarsoprol treatment appears to have diminished. Eflornithine seems to be a better first-line therapy for treating late-stage Gambian trypanosomiasis in the Republic of the Congo. PMID- 17128359 TI - Are health interventions implemented where they are most needed? District uptake of the integrated management of childhood illness strategy in Brazil, Peru and the United Republic of Tanzania. AB - OBJECTIVE: To describe geographical patterns of implementation of the Integrated Management of Childhood Illness (IMCI) strategy in three countries and to assess whether the strategy was implemented in areas with the most pressing child health needs. METHODS: We conducted interviews with key informants at the national and district levels in Brazil, Peru and the United Republic of Tanzania, and an ecological study of factors associated with health worker training in IMCI. Explanatory factors included district population, distance from the capital, human development index, other socioeconomic indicators and baseline mortality rates in children younger than five years. FINDINGS: In line with recommendations by WHO, early implementation districts were characterized by proximity to the capital and suitable training sites, presence of motivated health managers and a functioning health system. In the expansion phase, IMCI tended to be adopted by other districts with similar characteristics. In Brazil, uptake by poor and small municipalities and those further away from the state capital was significantly lower. In Peru, there was no association with distance from Lima, and a non significant trend for IMCI adoption by small and poor departments. In the United Republic of Tanzania, the only statistically significant finding was a lower uptake by remote districts. Implementation was not associated with baseline mortality levels in any country studied. CONCLUSION: Whereas clear and reasonable guidelines are provided for selection of early use districts, no criteria for promoting IMCI expansion had been issued, and areas of greatest need were not prioritized. Equity analyses based on the geographical deployment of new programmes and strategies can contribute to assessing whether they are reaching those who need them most. PMID- 17128360 TI - Ten-year health service use outcomes in a population-based cohort of 21,000 injured adults: the Manitoba injury outcome study. AB - OBJECTIVE: To quantify long-term health service use (HSU) following non-fatal injury in adults. METHODS: A retrospective, population-based, matched cohort study identified an inception cohort (1988-91) of injured people who had been hospitalized (ICD-9-CM 800-995) aged 18-64 years (n = 21 032) and a matched non injured comparison group (n = 21 032) from linked administrative data from Manitoba, Canada. HSU data (on hospitalizations, cumulative length of stay, physician claims and placements in extended care services) were obtained for the 12 months before and 10 years after the injury. Negative binomial and Poisson regressions were used to quantify associations between injury and long-term HSU. FINDINGS: Statistically significant differences in the rates of HSU existed between the injured and non-injured cohorts for the pre-injury year and every year of the follow-up period. After controlling for pre-injury HSU, the attributable risk percentage indicated that 38.7% of all post-injury hospitalizations (n = 25 183), 68.9% of all years spent in hospital (n = 1031), 21.9% of physician claims (n = 269 318) and 77.1% of the care home placements (n = 189) in the injured cohort could be attributed to being injured. CONCLUSION: Many people who survive the initial period following injury, face long periods of inpatient care (and frequent readmissions), high levels of contact with physicians and an increased risk of premature placement in institutional care. Population estimates of the burden of injury could be refined by including long term non-fatal health consequences and controlling for the effect of pre-injury comorbidity. PMID- 17128361 TI - Impact of conjugate Haemophilus influenzae type b (Hib) vaccine introduction in South Africa. AB - OBJECTIVE: To analyse trends in reported invasive Haemophilus influenzae disease in South Africa within the first five years of introduction of conjugate Haemophilus influenzae type b (Hib) vaccine in the routine child immunization schedule. METHODS: We used national laboratory-based surveillance data to identify cases of invasive H. influenzae disease between July 1999 and June 2004, and submitted isolates for serotyping and antimicrobial susceptibility testing. FINDINGS: The absolute number of Hib cases (reported to the national surveillance system) among children below one year of age decreased by 65%, from 55 cases in 1999-2000 to 19 cases in 2003-04. Enhanced surveillance initiated in 2003, identified human immunodeficiency virus (HIV)-infection and incomplete vaccination as contributing factors for Hib transmission. The total number of laboratory-confirmed cases of H. influenzae remained unchanged because non-type b disease was being increasingly reported to the surveillance system concomitant with system enhancements. Children with non-typable disease were more likely to be HIV-positive (32 of 34, 94%) than children with Hib disease (10 of 14, 71%), P = 0.051. Recent Hib isolates were more likely to be multidrug resistant (2% in 1999-2000 versus 19% in 2003-04, P = 0.001). CONCLUSION: Data from a newly established national laboratory-based surveillance system showed a decrease in Hib disease burden among South African children following conjugate vaccine introduction and identified cases of non-typable disease associated with HIV infection. PMID- 17128362 TI - Danger signs of neonatal illnesses: perceptions of caregivers and health workers in northern India. AB - OBJECTIVE: To assess household practices that can affect neonatal health, from the perspective of caregivers and health workers; to identify signs in neonates leading either to recognition of illness or health-care seeking; and to ascertain the proportion of caregivers who recognize the individual items of the integrated management of neonatal and childhood illnesses (IMNCI) programme. METHODS: The study was carried out in a rural community in Sarojininagar Block, Uttar Pradesh, India, using qualitative and quantitative research designs. Study participants were mothers, grandmothers, grandfathers, fathers or "nannies" (other female relatives) caring for infants younger than 6 months of age and recognized health care providers serving the area. Focus group discussions (n = 7), key informant interviews (n = 35) and structured interviews (n = 210) were conducted with these participants. FINDINGS: Many household practices were observed which could adversely affect maternal and neonatal health. Among 200 caregivers, 70.5% reported home deliveries conducted by local untrained nurses or relatives, and most mothers initiated breastfeeding only on day 3. More than half of the caregivers recognized fever, irritability, weakness, abdominal distension/vomiting, slow breathing and diarrhoea as danger signs in neonates. Seventy-nine (39.5%) of the caregivers had seen a sick neonate in the family in the past 2 years, with 30.38% in whom illness manifested as continuous crying. Health care was sought for 46 (23%) neonates. Traditional medicines were used for treatment of bulging fontanelle, chest in-drawing and rapid breathing. CONCLUSION: Because there is no universal recognition of danger signs in neonates, and potentially harmful antenatal and birthing practices are followed, there is a need to give priority to implementing IMNCI, and possible incorporation of continuous crying as an additional danger sign. PMID- 17128363 TI - Chemoprophylaxis and the epidemiological characteristics of re-emergent P. vivax malaria in the Republic of Korea. AB - OBJECTIVE: In the Republic of Korea (ROK), soldiers stationed where there is a risk of contracting malaria have received antimalarial chemoprophylaxis since 1997. However, chemoprophylaxis may facilitate the development of drug resistance, and late primary attacks in individuals who have received chemoprophylaxis are becoming more frequent. We investigated the association between chemoprophylaxis and the epidemiological characteristics and effectiveness of treatment for re-emergent Plasmodium vivax malaria, using a nationwide malaria database. METHODS: Among soldiers at risk of malaria between 1999 and 2001, we reviewed all P. vivax malaria cases (1158) that occurred before 31 December 2003. Early and late primary attacks were defined as cases occurring 2 months after the last day of exposure to risk of malaria, respectively. FINDINGS: Of these cases, 634 (72.0%) had received chemoprophylaxis, and 324 (28.0%) had not. Cases occurred mostly in summer, with a peak in July-August. Stratification by chemoprophylaxis history revealed different times to onset. Early primary attacks were more prevalent in the group not receiving chemoprophylaxis, while in the group receiving chemoprophylaxis most cases were late primary attacks. Of the latter, 312 out of 461 (67.7%) did not take primaquine regularly. After treatment of the first attack, 14 (1.2%) of 1158 were re-treated; all re-treated cases were cured using the same doses and regimen used for the first treatment. CONCLUSION: In ROK, the increase in late primary episodes of re-emergent P. vivax malaria is associated with the use of antimalarial chemoprophylaxis. PMID- 17128364 TI - The globalization of emergency medicine and its importance for public health. AB - Emergency medicine (EM) is a global discipline that provides secondary disease prevention and is also a tool for primary prevention. It is a horizontally integrated system of emergency care consisting of access to EM care; provision of EM care in the community and during transportation of patients; and provision of care at the receiving facility or hospital emergency department. EM can offer many tools to improve public health. These tools include primary disease prevention; interventions for addressing substance abuse and interpersonal violence; education about safety practices; epidemiological surveillance; enrolment of patients in clinical research trials focusing on acute interventions; education and clinical training of health-care providers; and participation in local and regional responses to natural and man-made disasters. Public health advocates and health policy-makers can benefit from the opportunities of EM and can help overcome its challenges. Advocating the establishment and recognition of the specialty of EM worldwide can result in benefits for health-care education, help in incorporating the full scope of EM care into the system of public health, and expand the capabilities of EM for primary and secondary prevention for the benefit of the health of the public. PMID- 17128365 TI - Adolescent suicide in the Middle East: ostrich head in sand. PMID- 17128366 TI - [Imaging in hepatosplenic candidiasis]. PMID- 17128376 TI - Reproducible Simulation of Respiratory Motion in Porcine Lung Explants. AB - PURPOSE: To develop a model for exactly reproducible respiration motion simulations of animal lung explants inside an MR-compatible chest phantom. MATERIALS AND METHODS: The materials included a piston pump and a flexible silicone reconstruction of a porcine diaphragm and were used in combination with an established MR-compatible chest phantom for porcine heart-lung preparations. The rhythmic inflation and deflation of the diaphragm at the bottom of the artificial thorax with water (1 - 1.5 L) induced lung tissue displacement resembling diaphragmatic breathing. This system was tested on five porcine heart lung preparations using 1.5T MRI with transverse and coronal 3D-GRE (TR/TE = 3.63/1.58, 256 x 256 matrix, 350 mm FOV, 4 mm slices) and half Fourier T2-FSE (TR/TE = 545/29, 256 x 192, 350 mm, 6 mm) as well as multiple row detector CT (16 x 1 mm collimation, pitch 1.5, FOV 400 mm, 120 mAs) acquired at five fixed inspiration levels. Dynamic CT scans and coronal MRI with dynamic 2D-GRE and 2D SS-GRE sequences (image frequencies of 10/sec and 3/sec, respectively) were acquired during continuous "breathing" (7/minute). The position of the piston pump was visually correlated with the respiratory motion visible through the transparent wall of the phantom and with dynamic displays of CT and MR images. An elastic body splines analysis of the respiratory motion was performed using CT data. RESULTS: Visual evaluation of MRI and CT showed three-dimensional movement of the lung tissue throughout the respiration cycle. Local tissue displacement inside the lung explants was documented with motion maps calculated from CT. The maximum displacement at the top of the diaphragm (mean 26.26 [SD 1.9] mm on CT and 27.16 [SD 1.5] mm on MRI, respectively [p = 0.25; Wilcoxon test]) was in the range of tidal breathing in human patients. CONCLUSION: The chest phantom with a diaphragmatic pump is a promising platform for multi-modality imaging studies of the effects of respiratory lung motion. PMID- 17128377 TI - Dark lumen MR colonography: can high spatial resolution VIBE imaging improve the detection of colorectal masses? AB - PURPOSE: To assess whether the detection of colorectal lesions can be improved using high spatial resolution VIBE imaging. MATERIALS AND METHODS: 48 patients underwent same-day dark lumen MR colonography (MRC) and conventional colonoscopy (CC) as the standard for the detection of colorectal masses. MRC was performed using contrast-enhanced standard and high spatial resolution T1-weighted 3D VIBE sequences. The findings and the image quality of the standard and high spatial resolution VIBE sequences were compared qualitatively and quantitatively. The findings of both sequences regarding colorectal lesions were compared to those of a subsequently performed colonoscopy. RESULTS: The high spatial resolution VIBE sequence significantly improved the quantitative image quality (CNR 54.0 vs. 36.8). However, high spatial resolution VIBE imaging did not detect more colorectal lesions than the standard VIBE sequence. In addition, none of the sequences employed was able to detect lesions with a diameter of less than 5 mm (CC 40 lesions). However, 13 colorectal lesions with a diameter of greater than 5 mm were detected by both sequences (CC 15). CONCLUSION: High spatial resolution VIBE imaging did not improve the detection of colorectal masses and MRC fails to detect colorectal lesions with a diameter of less than 5 mm. PMID- 17128378 TI - Experimental 16-row CT evaluation of in-stent restenosis using new stationary and moving cardiac stent phantoms: experimental examination. AB - PURPOSE: The aim of this study was to evaluate in-stent restenosiss using a newly developed stationary and moving cardiac stent phantom with three built-in artificial stenoses and a 16-row MDCT. MATERIALS AND METHODS: A newly developed coronary stent phantom with three artificial stenoses--low (approx. 30 %), medium (approx. 50 %) and high (approx. 70 %)--was attached to a moving heart phantom and used to evaluate the ability of 16-row MDCT to visualize in-stent restenosis. High resolution scans (16 x 0.75 mm, 250 mm FOV) were made to identify the baseline for image quality. The non-moving phantom was scanned (16 x 0.75 mm, routine cardiac scan protocol) first without and then with implementation of an ECG signal at various simulated heart rates (HR 40 to 120 bpm) and pitches (0.15 to 0.3). The moving cardiac phantom was scanned at the same simulated heart rates but at a pitch of 0.15. Images were reconstructed at every 10 % of the RR interval using a multi-cycle real cone-beam reconstruction algorithm. Multi planar reformations (MPR) were made for the image evaluation. The image quality was assessed using a three-point scale, and stent patency and stenoses detection were evaluated using a four-point scale. To evaluate the image quality and to grade the stent stenoses, the median values were calculated while considering the reconstruction interval. RESULTS: The image quality for the static phantom was adequate in 97 % of the measurements. In this phantom, every stenosis was detected independent of the pitch and heart rate used. The dynamic stent phantom yielded the best results at 0 %, 40 %, and 50 % of the RR interval at a pitch of 0.15. The low stenosis was visible at a simulated heart rate of up to 80 bpm. Patency can be detected at heart rates greater than 80 bpm. CONCLUSION: The newly developed moving stent phantom allowed a nearly in-vivo condition for detecting re-stenoses within a stent. In this phantom study the use of a 16-row MDCT allowed the detection of re-stenosis within a coronary stent at a heart rate of up to 80 bpm. This phantom can then be used for future studies, e. g. with a 64 row MDCT. PMID- 17128379 TI - [Infradiaphragmatic extralobar pulmonary sequestration]. AB - PURPOSE: To demonstrate characteristic imaging findings in infradiaphragmatic extralobar pulmonary sequestration (IEPS) with special emphasis on ultrasound (US). MATERIALS AND METHODS: The imaging material (pre- and postnatal US in all cases, magnetic resonance imaging (MRI) in 2 cases) for 4 infants (3 girls, 1 boy) was reviewed. 2 patients underwent surgery (after birth and at 4 months of age, respectively) and the diagnosis of IEPS was confirmed. The other 2 patients were monitored conservatively using US for up to 15 months. RESULTS: All 4 left side suprarenal masses exhibited the same characteristic sono-morphology, leading to the suspected diagnosis of IEPS. The masses were small (max. 10 ml), hyperechoic with cystic components and without calcifications, well demarcated and separate from the normal kidney and the suprarenal gland, and without any change in prenatal and directly postnatal size. Doppler US showed low-grade perfusion in all cases and an aberrant systemic artery originating from the abdominal aorta in 2 cases. MRI did not add any fundamental information. Despite the suspected imaging diagnosis of IEPS and negative urinalyses for neuroblastoma, 2 patients underwent surgery for histological confirmation. The lesions in the other 2 patients were monitored via US. A complete disappearance after 4 months in one patient and a continuous decrease in size over 15 months in the other patient were documented. CONCLUSION: Based on the characteristic findings of prenatal and postnatal US, IEPS can be diagnosed reliably. With the knowledge of its benign spontaneously regressing behavior, suspected suprarenal lesions should be treated conservatively via US monitoring. PMID- 17128380 TI - [Does interventional therapy prolong the patency of hemodialysis fistulas and grafts?]. AB - PURPOSE: To retrospectively evaluate procedural success and patency after endovascular treatment of acute dysfunctional hemodialysis fistulas and grafts in a non-preselected patient cohort. MATERIALS AND METHODS: 185 angiographies of hemodialysis fistulas and grafts on the upper extremities were analyzed for 120 patients (53 male, 67 female; mean-age 63.1 +/- 11.4, range 24 - 91). 70 % (n = 130) were native arteriovenous fistulas, 17 % (n = 31) were prosthetic grafts, and 13 % (n = 24) were non-specific. In total, 278 lesions requiring endovascular treatment were detected. 13 % (n = 35) of the lesions were located in the arterial inflow, 18 % (n = 49) in native arteriovenous anastomoses, 7 % (n = 19) in prosthetic grafts and 62 % (n = 171) in the venous outflow. The primary, secondary and cumulative patency after endovascular treatment was calculated. RESULTS: In 51 % (n = 94) of the cases endovascular treatment could be performed, in 8 % (n = 14) no lesion requiring treatment was detected, and in 42 % (n = 77) intervention was not considered possible. In 45 % (n = 124) of the detected lesions endovascular treatment was successful, in 18 % (n = 51) the intervention failed, and in 37 % (n = 103) intervention was not considered possible. The complication rate was 5 % (n = 10). The primary, secondary, and cumulative patency rates for 50 % of the hemodialysis fistulas and grafts after endovascular treatment were 65, 191, and 370 days, respectively. The results differed significantly from each other with p < 0.05 in the log rank test and log rank trend test. CONCLUSION: Endovascular treatment of acute dysfunctional hemodialysis fistulas and grafts is effective in restoring the patency for hemodialysis. PMID- 17128381 TI - [In vivo proton MR spectroscopy of normal liver parenchyma: technique and results]. AB - PURPOSE: To analyze the proton magnetic resonance spectroscopic data ( (1)H MRS) of normal liver parenchyma with regard to age, sex, body mass index and location in the liver. MATERIALS AND METHODS: 45 healthy volunteers age 24 to 65 years were examined with an optimized single-voxel (1)H MRS using a 1.5-T scanner. A spin echo sequence with a TR of 1500 ms and a TE of 135 ms was used, allowing in phase detection of the choline signal. Weak water suppression was achieved using a chemical shift selective suppression (CHESS) technique. Each examination included the measurement of three voxels with a voxel size of 18 x 18 x 18 mm (3) in different areas of the liver. The volunteers were divided into different age based groups (young: < or = 44 years; older: > or = 44 years), BMI (normal weighted: < 25 kg/m (2); obese: > 25 kg/m (2)) and sex. RESULTS: In the acquired spectra different lipid (e. g. [CH (2)] (n)), choline, glutamine, glutamate and glycogen-glucose-complex resonances were detected. The analysis of the spectra, however, only focused on the concentrations of choline and (CH (2)) (n) and the relative concentrations of the choline-to-(CH (2)) (n)-ratios. In the older volunteers the relative concentration of the choline-to-(CH (2)) (n)-ratio was significantly decreased by 0.213 +/- 0.193 in comparison to the younger subjects (p = 0,031). Further statistical analysis confirmed a significant decrease of the choline-to-(CH (2)) (n)-ratio by 0.223 +/- 0.180 in obese volunteers compared to volunteers of a standard weight (p = 0,016). The significant difference between the choline-to-(CH (2)) (n)-ratio in female versus male volunteers was calculated with an increase of 0.483 +/- 0.172 (p = 0,000). The location of the voxel in the liver parenchyma did not yield a significant difference in the choline-to-(CH (2)) (n)-ratio. CONCLUSION: The analysis of the proton liver MRS of healthy volunteers indicated a significant difference in the choline-to-(CH (2)) (n) ratio depending on age, sex, and BMI with a confidence interval of 95 %. The different choline-to-(CH (2)) (n)-ratio could be the result of the body fat distribution depending on age and sex and also of the increased fat portion of the body in obese volunteers. PMID- 17128382 TI - [Deep leg and pelvic vein thrombosis in a 4-year-old boy]. PMID- 17128383 TI - [Cherubism in CT diagnosis--a case report]. PMID- 17128384 TI - [Experimental Radiology 2006, Kiel, September 15-16, 2006. Abstracts]. PMID- 17128388 TI - Antibacterial and anticoagulant activities of coumarins isolated from the flowers of Magydaris tomentosa. AB - The phytochemical investigation of the acetone and methanol extracts of the flowers of Magydaris tomentosa (Desf.) DC afforded six known coumarins as well as (+)-meranzin hydrate (7), not previously reported as a natural product. The antibacterial activity of umbelliprenin (1), osthol (2), imperatorin (3), citropten (4) and (+)-meranzin hydrate (7) was tested against Gram-positive and Gram-negative bacteria. All coumarins (1-7) isolated in this study inhibited growth of all bacterial strains tested (MIC between 16 and 256 microg/mL), the most active being imperatorin (3) (MICs between 32 and 128 microg/mL) and citropten (4) (MICs between 16 and 256 microg/mL). The anticoagulant activity of compounds 1-4 and 7 was also evaluated. PMID- 17128389 TI - Molecular authentication of the traditional Chinese medicinal plant Euphorbia pekinensis. AB - The ITS regions of Euphorbia pekinensis and six other Euphorbia species used as adulterants of E. pekinensis were sequenced to differentiate them. The sequences are identical among the individuals in the seven species studies. Diversity in DNA sequences among various species was found ranging from 8.3% to 43.8% in ITS1 and 7.6% to 36.6% in ITS2 region. Furthermore, based on the divergent ITS regions, species-specific primers, JDJp 1 and JDJp 2, were designed in the polymorphic regions of E. pekinensis to distinguish it from adulterants. These ITS-derived primers amplified a 281-bp-specific DNA fragment from E. pekinensis. No amplified product was observed using DNA of six adulterants. PMID- 17128390 TI - Inhibitory activity of phenolic glycosides from the fruits of Idesia polycarpa on lipopolysaccharide-induced nitric oxide production in BV2 microglia. AB - Two new phenolic glycosides characterized as idesin salicylate (1) and 2 hydroxyphenol-1-O-beta-D-glucopyranosyl-(6-->1)- alpha-L-rhamnopyranoside (2) were isolated with four known compounds, idescarpin (3), idesin (4), 1-hydroxy-6 oxocyclohex-2-enecarboxylic acid methyl ester (5) and salirepin (6), from the fruits of IDESIA POLYCARPA. All six compounds significantly inhibited lipopolysaccharide-induced nitric oxide production in BV2 microglia at concentrations from 1 microM to 100 microM. PMID- 17128391 TI - [Social psychiatry in the field of contrasting context between psychiatry and Social Medicine]. AB - OBJECTIVE: There has been a discussion going on that Social Psychiatry has entered a state of crisis in terms of its socio-scientific roots. Little is known, however, about its relationship to Social Medicine. The question is whether Social Psychiatry, having grown apart from Sociology, has also lost its scientific relation to Social Medicine. METHOD: A systematic literature analysis of all works published in "Psychiatrische Praxis" - PP and "Das Gesundheitswesen" -GHW in the years 2004/2005 was done. All works concerning Social Psychiatry were analysed. The same procedure was applied to all abstracts, posters and presentations for the annual meetings of the German Society of Social Medicine and Prevention (DGSMP). RESULTS: 10 % off all articles published in GHW and 97 % in PP address issues of Social Psychiatry. Apart from similarities in terms of their theoretical, practical and institutional background and the research methods applied, there are a number of differences. CONCLUSIONS: Social Psychiatry has not lost its scientific relation to Social Medicine, however, the scientific cooperation between the two needs to be intensified. PMID- 17128392 TI - [Perspectives]. PMID- 17128393 TI - [For and against: integrated directorate of medical and nursing staff]. PMID- 17128394 TI - [Is Akinesia algera by Paul Julius Mobius (1891) as a coenasthetic appearance a still up-to-date phenomenon?]. AB - OBJECTIVE: Considering as example Akinesia algera - postulated in 1891 as a disease by Paul Julius Mobius, the relevance for diagnosing body-related mental disturbances will be demonstrated . METHOD: Relevant original works of Mobius and of some of his well known contemporaries for instance Kraepelin, Binswanger, Erb were explored. The correlation of the described phenomenon to current textbook doctrine has been attempted. RESULTS: Mobius, case histories, described as Akinesia algera, can be subsumed under the typology of coenasthetic schizophrenia. CONCLUSION: Akinesia algera is by primary source to be evaluated as a coenasthetic symptom and remains in daily practice a diagnostic challenge. PMID- 17128395 TI - [Psychoeducation in schizophrenic disorders--psychotherapy or "infiltration"?]. AB - OBJECTIVE: Assumptions and applications of psychoeducational group treatment are critically reviewed, taking into account recent research and examining exemplarily the concept and procedures of the Arbeitsbuch PsychoEdukation bei Schizophrenie (APES) by Bauml et al. Further, the impact of the concept of illness and perception of man on psychiatric care and therapeutic relationships are discussed. METHODS: The assertions and rationale of psychoeducation are compared to its implementation in treatment and are related to the practice of care. RESULTS: Contrary to its goals, such as self competence and the ability to cope with crises, psychoeducation focuses on the neurobiological principles of dopaminergic dysfunction. Within a deficit model of psychiatric illness, individuals learn to accept the "stroke of fate of psychosis". Their contribution to recovery is basically reduced to improved medication compliance. CONCLUSIONS: Psychoeducation in the form of compliance training is not psychotherapy. Without involvement of family members it is also not very effective. A collaboration with health care professionals requires patients' self competence and the proper utilization of resources. PMID- 17128396 TI - [Vocational therapy in schizophrenia: effects only with some patients and with which patients?]. AB - OBJECTIVES: In a previous study the effectiveness of vocational therapy was evaluated using the Osnabruck Profile of Working Abilities (O-AFP). No or only minor effects were detected. The goal of this reanalysis is to identify distinct responder groups. METHODS: Hierarchical cluster analysis of O-AFP-scores before and after treatment, anovas, t-statistics as well as regression and discriminant analyses were applied to specify the clusters. RESULTS: For each of the O-AFP scales (Learning Ability, Social Communication Ability, Adaptation) three subgroups different in level and slope of abilities could be identified. Improvements with medium to high effect sizes were detected for Social Communication Ability (34 %) and Learning Ability (14 % of the patients), deterioration was found in 30 % of the patients for Adaptation. Subgroups could be described with the help of symptomatology. Further variables and subgroup membership could be predicted above chance by discriminant and regression analysis. CONCLUSION: Targeted and more appropriate design of vocational therapy seems necessary. PMID- 17128397 TI - [Paranoid schizophrenia in monozygotic twins--from caregiver to patient affected]. AB - OBJECTIVE: The aim of this case study is to show how a monozygotic twin perceives the schizophrenic disorder of his twin brother and how these experiences affect his own handling and coping after the beginning of his own schizophrenic disorder. PATIENT: A 46-year-old patient with a first episode of schizophrenia who took care of his twin brother with a disease history of 25 years of schizophrenia. RESULTS: In this case the little knowledge the patient had of his schizophrenic disorder and his lasting lack accessing his illness in the context of a defensive mechanism were striking. Concurrently, the patient overestimated his loading capacity. CONCLUSIONS: A long time of caregiving of a schizophrenic twin brother does not necessarily facilitate the coping with a schizophrenic illness of one's own. Defensive mechanisms could inhibit the implementation of constructive coping strategies with the illness. PMID- 17128398 TI - [Dilemmas of expert opinion and coercive treatment according to section sign 64 German penal code]. AB - AIMS: Compulsory admissions according to section sign 64 German penal code are rising. There is also a dramatic increase in dropout-rates with regard to coercive therapeutic interventions in forensic facilities specialized in the treatment of offenders addicted to drugs or alcohol. METHODS: By means of two case histories dilemmas of expert opinion and coercive treatment of offenders addicted to drugs or alcohol are illustrated. RESULTS AND CONCLUSIONS: Consideration of current results in addiction research should be an obligatory element of expert opinion and coercive treatment programmes in forensic facilities to provide for sufficient quality standards. Even when strict quality criteria are considered failures in placement of offenders with drug or alcohol addiction can not be ruled out. Besides scientific criteria a variety of other factors may influence the decision to coercive drug addiction therapy in a forensic facility e. g. jurisdiction or even tactical statements of the offender. PMID- 17128399 TI - [Therapy and assessment of psychological trauma as mirrored by fiction--part 1: World War I in British perspective]. PMID- 17128402 TI - [Regarding the article, Are we still in need of psychiatry as a special field within medicine]. PMID- 17128403 TI - [Regarding the article, Involuntary hospitalisation in 2000 according to German, PsychKG in the City of Hannover]. PMID- 17128405 TI - Modulation of apelin and APJ receptor in normal and preeclampsia-complicated placentas. AB - Apelin is an endogenous ligand of the human orphan receptor APJ. This peptide is produced through processing from the C-terminal portion in the pre-pro-protein consisting of 77 amino acid residues and exists in multiple molecular forms. Although the main physiological functions of apelin have not yet been clarified, it is known that apelin is involved in the regulation of blood pressure, blood flow and central control of body fluid homeostasis in different organs. Since human placenta is a tissue where vasculogenesis, blood pressure and flow are dramatically important to allow a normal embryonic and fetal growth and development, the aim of the present study was to investigate the immunohistochemical distribution of apelin and APJ in normal placentas throughout pregnancy and in preeclampsia-complicated placentas. Specifically, we observed that in normal placentas the expression levels of apelin decreased from the first to the third trimester of gestation in both cytotrophoblast and syncytiotrophoblast cells and in the stroma of placental villi, in contrast with increased expression levels of APJ in the cytoplasm of cytotrophoblast cells and in the cytoplasm of endothelial cells of normal placenta samples. In contrast, in preeclampsia-complicated pregnancies, we observed a very strong increase of expression levels of both apelin and APJ receptor in all the placental compartments, cytotrophoblast, syncytiotrophoblast and stroma with a particular increase in endothelial cells inside preeclamptic placental villi. Our data seem to indicate an important role of apelin and APJ in the regulation of fetal development through a correct regulation of human placenta formation during pregnancy. Moreover, the strong expression levels of apelin and APJ in preeclamptic placentas, suggest their possible involvement in the onset of this pathology. PMID- 17128406 TI - Mucoepidermoid tumors of the bronchus. Ultrastructural and immunohistochemical study. Histiogenic correlations. AB - Bronchial mucoepidermoid tumors are uncommon neoplasms, morphologically similar to their salivary gland counterpart. The histogenesis is controversial. The aim of this study is to identify myoepithelial cells and speculate on their role in the origin of these tumors. METHODS AND RESULTS: Sixteen bronchial mucoepidermoid tumor surgical specimens were formalin-fixed, paraffin-embedded and studied using a panel of nine antibodies in order to identify a myoepithelial differentiation. Additional antigens against several cytokeratins were performed in four cases and five of the biopies were studied using the electron microscopy. The different types of cells of the primary bronchial mucoepidermoid tumor (mucous luminal, intermediate and squamous) reacted strongly against AE1, CK7, 34bE12 and weakly with AE3, CK18 and CK8/18/19. S-100, alpha-smooth muscle actin, muscle actin HHF35 and alpha-actinin were consistently negative in all cell types. CD10 was positive in very few cells in just one case. CONCLUSION: The immunohistochemical and the ultrastructural study of bronchial mucoepidermiod tumors support a ductal unit origin, without a myoepithelial participation. PMID- 17128407 TI - Phthalate esters immunolocalized in the gastrointestinal tract of shi drum Umbrina cirrosa (L.) and rainbow trout, Oncorhynchus mykiss (W.). AB - The occurrence of phthalate esters in freshwater and marine aquacultural species like rainbow trout Oncorhynchus mykiss and shi drum Umbrina cirrosa, respectively, were determined by immunohistochemical approach. The results showed a similar distribution in the gastrointestinal tract of both species. In particular, intense immunoreactivity was found at gastric gland level. In the intestinal tract, goblet cells failed to stain, whereas enterocytes showed the highest binding of phthalates restricted to the apical cytoplasm. This distribution of phthalate esters at gastric gland and enterocyte level may have implications for the physiology of the digestive process and intestinal biotransformation. Phthalates are confirmed to be widely diffused contaminants, absorbed via the alimentary canal; thus a multidisciplinary approach could be useful to examine sea and freshwater environments. PMID- 17128408 TI - Histochemical study of glycoconjugates in the toadfish Halobatrachus didactylus oesophagus epithelium. AB - The carbohydrate expression in the epithelium lining the oesophagus of the toadfish Halobatrachus didactylus was studied by means of conventional and lectin histochemistry. The stratified epithelium was constituted by basal cells, polymorphous cells in the intermediate layer, pyramidal and flattened cells in the outer layer and contained two types of large secretory cells: goblet cells and sacciform cells. PAS, Alcian blue pH 2.5 and pH 1.0 stained very strongly the goblet cells, weakly the surface of the other epithelial cells but did not stain the sacciform cells. The goblet cells cytoplasm contained oligosaccharides with terminal Galbeta1,3GalNAc, alpha/betaGalNAc, Galbeta1,4GlcNAc, alphaL-Fuc and internal betaGlcNAc residues (PNA, SBA, RCA120, UEA I, LTA and KOH-sialidase-WGA affinity). Galbeta1,4GlcNAc, alphaL-Fuc and internal betaGlcNAc were also found in the glycocalyx. The sacciform cells expressed sialyloligosaccharides terminating with Neu5Acalpha2,3Galbeta1,4GlcNac, Neu5Acbeta2,6Gal/GalNAc, Neu5AcForssman pentasaccharide (MAL II, SNA, KOH-sialidase-DBA staining) as well as asialo-glycoconjugates with terminal/internal alphaMan (Con A affinity) and with terminal Galbeta1,3GalNAc, Forssman pentasaccharide, Galbeta1,4GlcNAc, GalNAc (HPA and SBA reactivity), alphaGal (GSA I-B4 reactivity), D-GlcNAc (GSA II labelling), alphaL-Fuc. The basal cells cytoplasm exhibited terminal/internal alphaMan and terminal Neu5Acalpha2,6Gal/GalNAc, Galbeta1,4GlcNAc, alpha/betaGalNAc, alphaGal, GlcNAc, alphaL-Fuc. Intermediate cells showed oligosaccharides with terminal/internal alphaMan and/or terminating with Neu5Acalpha2,6Gal/GalNAc, Galbeta1,4GlcNAc in the cytoplasm and with Neu5Acalpha2,3Galbeta1,4GlcNac, alpha/betaGalNAc, alphaGal, GlcNAc, alphaL-Fuc in the glycocalyx. The pyramidal cells expressed terminal/internal alphaMan and terminal Neu5Acalpha2,6Gal/GalNAc, alpha/betaGalbeta1,4NAc, alphaGal, alphaL-Fuc in the entire cytoplasm, terminal Neu5Acalpha2,3Galbeta1,4GlcNac and Forssman pentasaccharide in the apical extension, internal betaGlcNAc and/or terminal alphaL-Fuc in the luminal surface, Neu5Acalpha2,3Galbeta1,4GlcNac, Neu5Acalpha2,6Gal/GalNAc, Galbeta1,4GlcNAc, alphaGal in the basolateral surface. The flattened cells displayed glycans with terminal/internal alphaMan and terminal Neu5Acalpha2,6Gal/GalNAc, alpha/betaGalNAc, alphaGal, D-GlcNAc in the entire cytoplasm, glycans terminating with Galbeta1,3GalNAc and/or internal betaGlcNAc in the sub-nuclear cytoplasm. PMID- 17128409 TI - Comparative cytokeratin distribution patterns in cholesteatoma epithelium. AB - Cytokeratins (CKs) are known as the intermediate filament proteins of epithelial origin. Their distribution in human epithelia is different according to the type of epithelium, state of growth and differentiation. We used monoclonal mouse antibodies against cytokeratins to study CK expression in the following human tissues: cholesteatoma, middle ear mucosa, glandular epithelium, and meatal ear canal epithelium. Immunohistochemical processing was performed using the labeled steptavidin peroxidase method to demonstrate the presence of CKs in cells of human epidermis. Positive reaction was obtained for CK4, CK34betaE12, CK10, CK14 in skin and cholesteatoma epithelium. However, a more extensive positive reaction with those CKs was observed in cholesteatoma epithelium. Positive immunoreactivity was seen with anti- CK19 in the glandular epithelium. Middle ear mucosa specimens revealed positive immunoreactivity with the antibodies against CK4. The expression of CK4 was definitely positive within the basal layers of the epidermis. The glandular epithelium showed no positive reaction with anti- CK4, anti- CK34betaE12, anti- CK14 and anti-CK10. Immunohistochemistry for CK18 showed no reaction in all examined tissues. Cholesteatoma is known as a proliferative disease in the middle ear which pathogenesis is not completely understood. Keratinocytes express hyperproliferation- associated CKs and after reaching the suprabasal layers they finally undergo apoptosis creating keratinous debris. Cytokeratin expression observed in the epithelium explains proliferative behavior of cholesteatoma which is associated with increased keratinocyte migration. Cytokeratins can be used as potential proliferative markers. It can also allow for searching the usefulness of inhibiting regulators in the treatment of hyperproliferative diseases. PMID- 17128410 TI - Airway and lung parenchyma morphology during the respiratory cycle. AB - OBJECTIVE: Describe the morphological changes that take place in the lung parenchyma and in the airways during the respiratory cycle with a view to establishing a relationship between them. SUBJECTS: Adult Wistar rats. INTERVENTIONS: The lungs were fixed at seven different points in the respiratory cycle: Inflation, 10 and 20 cm. transpulmonary pressure, total lung capacity. Deflation, 20, 15, 10 and 0 cm transpulmonary pressure. MEASUREMENTS: The lungs were processed for morphometric study and bronchial and parenchymal variables, such as lung volume, number of alveoli, anatomic dead space, bronchial lumen surface and bronchial wall surface were quantified. The results were compared by analysis of variance (ANOVA) or the Kruskal-Wallis and Mann-Whitney's U tests. RESULTS: The lung volume, the number of alveoli and the anatomic dead space increased with the increase of the transpulmonary pressure and decreased with the decrease of it, the obtained values in deflation being higher than those in inflation (p<0.05). The bronchial lumen and the bronchial wall surfaces generally showed higher values in inflation than in deflation (p<0.05). CONCLUSIONS: The anatomic dead space was altered as a consequence of the variations in airway diameter and length. Lung parenchyma tension may have been of influence in the variations of the bronchial wall. PMID- 17128411 TI - Development of the mouse mandibles and clavicles in the absence of skeletal myogenesis. AB - In this report we employed double-knock-out mouse embryos and fetuses (designated as Myf5-/-: MyoD-/- that completely lacked striated musculature to study bone development in the absence of mechanical stimuli from the musculature and to distinguish between the effects that static loading and weight-bearing exhibit on embryonic development of skeletal system. We concentrated on development of the mandibles (= dentary) and clavicles because their formation is characterized by intramembranous and endochondral ossification via formation of secondary cartilage that is dependent on mechanical stimuli from the adjacent musculature. We employed morphometry and morphology at different embryonic stages and compared bone development in double-mutant and control embryos and fetuses. Our findings can be summarized as follows: a) the examined mutant bones had significantly altered shape and size that we described morphometrically, b) the effects of muscle absence varied depending on the bone (clavicles being more dependent than mandibles) and even within the same bone (e.g., the mandible), and c) we further supported the notion that, from the evolutionary point of view, mammalian clavicles arise under different influences from those that initiate the furcula (wishbone) in birds. Together, our data show that the development of secondary cartilage, and in turn the development of the final shape and size of the bones, is strongly influenced by mechanical cues from the skeletal musculature. PMID- 17128412 TI - Survivin and Cyclooxygenase-2 are co-expressed in human and mouse colon carcinoma and in terminally differentiated colonocytes. AB - In the evolution of colon rectal cancer (CRC) the imbalance between cell proliferation and apoptosis is considered one of the prominent causes of tumor induction and/or progression. In order to establish the role of anti apoptotic proteins in colon cancer development, we studied with immunohistochemical techniques the expression of Survivin in a mouse model of colon carcinogenesis induced by 1,2-dimethyl-hydrazine treatment. In this mouse model Survivin was over-expressed during tumor development, showing a distribution mimicking that described in the correspondent human malignancies. We also correlated Survivin distribution with COX-2 and beta-Catenin expression patterns. The co-localization of COX-2/beta-Catenin/Survivin in the same epithelial cells in tumor samples lends credence to possible in vivo regulatory effects of COX-2 and beta-Catenin on the intracellular Survivin levels in mouse and human colon cancer. PMID- 17128413 TI - Fine structure of spermatozoa in the gilthead sea bream (Sparus aurata Linnaeus, 1758) (Perciformes, Sparidae). AB - Scanning and transmission electron microscopy were used to investigate the fine structure of the sperm of the sparid fish Sparus aurata L. The mature spermatozoon of gilthead sea bream belongs, like that of the other sparid fish, to a "type I" as defined by Mattei (1970). It has a spherical head which lacks an acrosome, a short, irregularly-shaped midpiece and a long cylindrical tail. The nucleus reveals a deep invagination (nuclear fossa) in which the centriolar complex is located. The two centrioles are approximately perpendicular to each other and show a conventional "9+0" pattern. The proximal centriole is associated with a cross-striated cylindrical body lying inside a peculiar satellite nuclear notch which appears as a narrow invagination of the nuclear fossa. The distal centriole is attached to the nuclear envelope by means of a lateral plate and radial fibres made of an electron-dense material. The short midpiece houses one mitochondrion. The flagellum is inserted perpendicularly into the base of the nucleus and contains the conventional 9+2 axoneme. PMID- 17128414 TI - Protection from oxidative stress by enhanced glycolysis; a possible mechanism of cellular immortalization. AB - Reactive oxygen species (ROS) play a crucial role not only in the physiological signal transduction but also in the pathogenesis of several human diseases such as atherosclerosis, neuro-degenerative diseases, metabolic disorders, aging or cancer amongst others. Oxidative stress is also responsible for cellular and organism senescence, in accordance with what Harman initially proposed in the free radical theory of aging. Recent findings support the notion that protection from oxidative stress can increase life span significantly. We reported that enhanced glycolysis could modulate cellular life span with reduction of oxidative stress. Moreover, the tumor suppressor gene p53 controls post-transcriptionally the level of the glycolytic enzyme, phosphoglycerate mutase (PGM). As enhanced glycolysis is a distinctive and prominent feature of cancer cells (termed the Warburg effect), our findings disclosed a novel aspect of the Warburg effect: the connection between senescence and oxidative stress. PMID- 17128415 TI - Emerging significance of ER-coregulator PELP1/MNAR in cancer. AB - The estrogen receptors ERalpha and ERbeta have been implicated in the progression of a wide variety of cancers. The actions of ER are regulated by ER coregulator proteins, including proline-, glutamic acid- and leucine-rich-protein-1 (PELP1/MNAR). PELP1 has been shown to participate in both genomic and nongenomic functions of ER. The expression and localization of PELP1/MNAR are deregulated in a wide variety of tumors and have been implicated in the development of hormonal resistance in cancer cell lines. Emerging data suggest that PELP1/MNAR interacts with many proteins and activates several oncogenes, including Src kinase, phosphotidyl inositol 3 kinase (PI3K), and signal transducers and activators of transcription 3 (STAT3). These new results suggest that PELP1/MNAR may act as an oncogene as well as cooperating with other oncogenes. Thus, PELP1/MNAR may contribute to the tumorigenic potential of cancer cells by serving as a scaffolding protein that couples various signaling complexes with ER. PMID- 17128416 TI - A review of FGF18: Its expression, signaling pathways and possible functions during embryogenesis and post-natal development. AB - FGF18 is a novel growth factor first reported in 1998. Current evidence suggests that FGF18 may play a prominent role in chondrogenesis and osteogenesis during skeletal development and growth. However, its function extends to many other biological processes. Although there remains much to be discovered and investigated on the functions and mechanisms of FGF18, it may play a role as a useful therapeutic target for various applications. The following review summarizes the current knowledge on FGF18 with special emphasis on its skeletal functions and highlights its potential areas for future research. PMID- 17128417 TI - Histopathological changes induced by therapies in the benign prostate and prostate adenocarcinoma. AB - The effect of androgen deprivation and other hormonal therapies, radiation therapy, thermal ablation therapies, chemotherapy, and other systemic treatments is evident in the histology of non-neoplastic and neoplastic human prostate gland. Androgen deprivation may be achieved with: a. orchidectomy, b. exogenous oestrogen administration, c. drugs with the capacity to deplete the hypothalamus of luteinizing hormone-releasing hormone, d. antiandrogens administration: drugs, which block the conversion of testosterone to its active form of 5-alpha dihydrotestosterone (i.e. finasteride, dutasteride), and drugs which block the androgen receptor on individual cells (i.e. flutamide). Androgen deprivation therapies cause atrophy of non-neoplastic and neoplastic prostatic epithelium, as the result of apoptosis, and are mainly used as a palliative measure in metastatic prostate cancer or as neoadjuvant or adjuvant treatment, in clinically localized prostate cancer. Morphological tumour regression may complicate the recognition and grading of treated carcinomas in radical prostatectomy specimens. Radiation therapy may be applied in the form of external beam, interstitial implantation (brachytherapy), or a combination, as a mainstay or adjuvant (external beam) treatment in localized prostate cancer. The primary effect is the damage of endothelial cells, which cause ischemia that leads to atrophy. The difficulty of post-radiation prostate needle biopsy interpretation includes the distinction of treatment effect in normal prostatic tissue from recurrent or residual tumour. Histological changes after thermal ablation mainly include lesions observed in prostatic infarcts due to periurethral coagulative type necrosis of variable volume. The correlation between the histopathological effects of the above therapies and their clinical significance is not absolutely clear. PMID- 17128418 TI - Inhibition of Flt3-activating mutations does not prevent constitutive activation of ERK/Akt/STAT pathways in some AML cells: a possible cause for the limited effectiveness of monotherapy with small-molecule inhibitors. AB - The Flt3 receptor tyrosine kinase is a critical mediator in the pathogenesis of acute myeloid leukaemia (AML). Flt3-activating mutations have been associated with poor prognosis and decreased overall survival of AML patients, thus Flt3 constitutes an ideal target for drug treatment of such disease. Unfortunately, the monotherapy with small-molecule tyrosine kinase inhibitors in clinical trials shows that remission is not permanent, presumably by resistance of Flt3 mutants to inhibitors. An alternative approach for treatment is based on the cooperation between Flt3 and additional intracellular pathways for AML transformation in some patients. Thus, the inhibition of both Flt3 and such pathways may be exploited for successful treatment of the disease. We investigated the importance of Flt3 activating mutations for the constitutive activation of intracellular pathways in primary AML cells, and their effect on cell survival. We found that the main compounds involved in the differentiation, proliferation and survival of AML (MAPK/AKT/STAT) were constitutively activated. However, only four samples showed internal tandem duplications (ITDs) for Flt3. Surprisingly, contrary to previous reports, we found that inhibition of ITD/Flt3 activity did not prevent the phosphorylation of ERK, STAT5 or Akt in some primary AML cells. In parallel, we found that in these cells, Flt3 and ERK or Akt cooperate to regulate cell survival. Our results support the hypothesis that the optimal therapeutic treatment of AML may require not only the oncogenic tyrosine kinase, but also the appropriate combination of different specific inhibitors, thus providing a more effective approach to reverse leukaemogenesis. Thus, we propose that each AML patient should have an individually tailored combination treatment. PMID- 17128419 TI - Amphiphilic poly(N-propargylamide) with galactose and lauryloyl groups: synthesis and properties. AB - An amphiphilic poly(N-propargylamide) with galactose and lauryloyl groups was synthesized by copolymerization of the corresponding N-propargylamide monomers using a Rh catalyst. The obtained copolymer formed a one-handed helical conformation and molecular aggregates in water. The observations by fluorescence microscopy in a cell culture experiment in the presence of dye-labeled copolymer indicated that the copolymer was incorporated into the cells. PMID- 17128420 TI - PEG-PLA block copolymer as potential drug carrier: preparation and characterization. AB - Diblock and multiblock copolymers composed of a poly(D,L-lactide) (PLA) or poly(trimethylene carbonate) (PTMC) core with a hydrophilic chain of poly(ethylene glycol) (PEG) were prepared. These copolymers, in which the core is connected to PEG through a polyfunctional molecule such as citric, mucic, or tartaric acid, may be used to form nanoparticles for drug delivery applications. Branched copolymers were prepared by direct amidation between the polyfunctional acid and methoxy PEGamine, followed by ring-opening polymerization of lactide or trimethyl carbonate to form the PLA and PTMC block copolymers. In addition, a complex multiblock copolymer of biotin-PEG-poly[lactic-co-(glycolic acid)] (PLGA) for application in an avidin-biotin system was prepared for possible design of nanospheres with targeting properties. Studies of drug release from polymeric systems containing multiblock copolymers and studies of polymer degradation were also performed. PMID- 17128421 TI - Thermo-responsive chitosan-graft-poly(N-isopropylacrylamide) injectable hydrogel for cultivation of chondrocytes and meniscus cells. AB - A thermo-responsive comb-like polymer with chitosan as the backbone and pendant poly(N-isopropylacrylamide) (PNIPAM) groups has been synthesized by grafting PNIPAM-COOH with a single carboxy end group onto chitosan through amide bond linkages. The copolymer exhibits reversible temperature-responsive soluble insoluble characteristics with the lower critical solution temperature (LCST) being at around 30 degrees C. Results from SEM observations confirm a porous 3D hydrogel structure with interconnected pores ranging from 10 to 40 microm at physiological temperature. A preliminary in vitro cell culture study has demonstrated the usefulness of this hydrogel as an injectable cell-carrier material for entrapping chondrocytes and meniscus cells. The hydrogel not only preserves the viability and phenotypic morphology of the entrapped cells but also stimulates the initial cell-cell interactions. PMID- 17128422 TI - Biodegradable injectable in situ depot-forming drug delivery systems. AB - The scope of drug-delivery systems has expanded significantly in recent years providing new ways to deliver life saving therapeutics to patients. The development of new injectable drug-delivery systems has provided new vistas and opened up unexplored horizons in the field of science, particularly in controlled drug delivery since these systems possess unique advantages over traditional ones, which include ease of application, and localized and prolonged drug delivery. In the past few years, an increasing number of such systems has been reported in the literature for various biomedical applications, including drug delivery, cell encapsulation, and tissue repair. These are injectable fluids that can be introduced into the body in a minimally invasive manner prior to solidifying or gelling within the desired site. For this purpose both natural (chitosan, alginates) as well as synthetic polymers (PEGylated polyesters, ricinoleic acid-based polymers) have been utilized. These systems have been explored widely for the delivery of various therapeutic agents ranging for anti neoplastic agents like paclitaxel to proteins and peptides such as insulin, almost covering every segment of the pharmaceutical field. This manuscript focuses on the recent advancements in the area of in situ forming biodegradable polymeric drug-delivery systems. PMID- 17128423 TI - Stimuli-responsive hydrogels based on polysaccharides incorporated with thermo responsive polymers as novel biomaterials. AB - In recent years, intelligent hydrogels which can change their swelling behavior and other properties in response to environmental stimuli such as temperature, pH, solvent composition and electric fields, have attracted great interest. The hydrogels based on polysaccharides incorporated with thermo-responsive polymers have shown unique properties such as biocompatibility, biodegradability, and biological functions in addition to the stimuli-responsive characters. These "smart" hydrogels exhibit single or multiple stimuli-responsive characters which could be used in biomedical applications, including controlled drug delivery, bioengineering or tissue engineering. This review focuses on the recent developments and future trends dealing with stimuli-responsive hydrogels based on grafting/blending of polysaccharides such as chitosan, alginate, cellulose, dextran and their derivatives with thermo-sensitive polymers. This review also screens the current applications of these hydrogels in the fields of drug delivery, tissue engineering and wound healing. PMID- 17128424 TI - A note on the power of Fisher's least significant difference procedure. AB - Fisher's least significant difference (LSD) procedure is a two-step testing procedure for pairwise comparisons of several treatment groups. In the first step of the procedure, a global test is performed for the null hypothesis that the expected means of all treatment groups under study are equal. If this global null hypothesis can be rejected at the pre-specified level of significance, then in the second step of the procedure, one is permitted in principle to perform all pairwise comparisons at the same level of significance (although in practice, not all of them may be of primary interest). Fisher's LSD procedure is known to preserve the experimentwise type I error rate at the nominal level of significance, if (and only if) the number of treatment groups is three. The procedure may therefore be applied to phase III clinical trials comparing two doses of an active treatment against placebo in the confirmatory sense (while in this case, no confirmatory comparison has to be performed between the two active treatment groups). The power properties of this approach are examined in the present paper. It is shown that the power of the first step global test--and therefore the power of the overall procedure--may be relevantly lower than the power of the pairwise comparison between the more-favourable active dose group and placebo. Achieving a certain overall power for this comparison with Fisher's LSD procedure--irrespective of the effect size at the less-favourable dose group- may require slightly larger treatment groups than sizing the study with respect to the simple Bonferroni alpha adjustment. Therefore if Fisher's LSD procedure is used to avoid an alpha adjustment for phase III clinical trials, the potential loss of power due to the first-step global test should be considered at the planning stage. PMID- 17128425 TI - Randomization as a basis for inference in noninferiority trials. AB - Noninferiority testing in clinical trials is commonly understood in a Neyman Pearson framework, and has been discussed in a Bayesian framework as well. In this paper, we discuss noninferiority testing in a Fisherian framework, in which the only assumption necessary for inference is the assumption of randomization of treatments to study subjects. Randomization plays an important role in not only the design but also the analysis of clinical trials, no matter the underlying inferential field. The ability to utilize permutation tests depends on assumptions around exchangeability, and we discuss the possible uses of permutation tests in active control noninferiority analyses. The other practical implications of this paper are admittedly minor but lead to better understanding of the historical and philosophical development of active control noninferiority testing. The conclusion may also frame discussion of other complicated issues in noninferiority testing, such as the role of an intention to treat analysis. PMID- 17128426 TI - Assessment of futility in clinical trials. AB - The term 'futility' is used to refer to the inability of a clinical trial to achieve its objectives. In particular, stopping a clinical trial when the interim results suggest that it is unlikely to achieve statistical significance can save resources that could be used on more promising research. There are various approaches that have been proposed to assess futility, including stochastic curtailment, predictive power, predictive probability, and group sequential methods. In this paper, we describe and contrast these approaches, and discuss several issues associated with futility analyses, such as ethical considerations, whether or not type I error can or should be reclaimed, one-sided vs two-sided futility rules, and the impact of futility analyses on power. PMID- 17128427 TI - Active-controlled, non-inferiority trials in oncology: arbitrary limits, infeasible sample sizes and uninformative data analysis. is there another way? AB - In oncology, it may not always be possible to evaluate the efficacy of new medicines in placebo-controlled trials. Furthermore, while some newer, biologically targeted anti-cancer treatments may be expected to deliver therapeutic benefit in terms of better tolerability or improved symptom control, they may not always be expected to provide increased efficacy relative to existing therapies. This naturally leads to the use of active-control, non inferiority trials to evaluate such treatments. In recent evaluations of anti cancer treatments, the non-inferiority margin has often been defined in terms of demonstrating that at least 50% of the active control effect has been retained by the new drug using methods such as those described by Rothmann et al., Statistics in Medicine 2003; 22:239-264 and Wang and Hung Controlled Clinical Trials 2003; 24:147-155. However, this approach can lead to prohibitively large clinical trials and results in a tendency to dichotomize trial outcome as either 'success' or 'failure' and thus oversimplifies interpretation. With relatively modest modification, these methods can be used to define a stepwise approach to design and analysis. In the first design step, the trial is sized to show indirectly that the new drug would have beaten placebo; in the second analysis step, the probability that the new drug is superior to placebo is assessed and, if sufficiently high in the third and final step, the relative efficacy of the new drug to control is assessed on a continuum of effect retention via an 'effect retention likelihood plot'. This stepwise approach is likely to provide a more complete assessment of relative efficacy so that the value of new treatments can be better judged. PMID- 17128428 TI - Sample size and the probability of a successful trial. AB - This paper describes the distinction between the concept of statistical power and the probability of getting a successful trial. While one can choose a very high statistical power to detect a certain treatment effect, the high statistical power does not necessarily translate to a high success probability if the treatment effect to detect is based on the perceived ability of the drug candidate. The crucial factor hinges on our knowledge of the drug's ability to deliver the effect used to power the study. The paper discusses a framework to calculate the 'average success probability' and demonstrates how uncertainty about the treatment effect could affect the average success probability for a confirmatory trial. It complements an earlier work by O'Hagan et al. (Pharmaceutical Statistics 2005; 4:187-201) published in this journal. Computer codes to calculate the average success probability are included. PMID- 17128429 TI - Inference methods for saturated models in longitudinal clinical trials with incomplete binary data. AB - In the longitudinal studies with binary response, it is often of interest to estimate the percentage of positive responses at each time point and the percentage of having at least one positive response by each time point. When missing data exist, the conventional method based on observed percentages could result in erroneous estimates. This study demonstrates two methods of using expectation-maximization (EM) and data augmentation (DA) algorithms in the estimation of the marginal and cumulative probabilities for incomplete longitudinal binary response data. Both methods provide unbiased estimates when the missingness mechanism is missing at random (MAR) assumption. Sensitivity analyses have been performed for cases when the MAR assumption is in question. PMID- 17128430 TI - In vitro antimicrobial activity of Zingiber cassumunar (Plai) oil and a 5% Plai oil gel. AB - The essential oil of Zingiber cassumunar (Plai oil) exhibits antimicrobial activity against a wide range of Gram-positive and Gram-negative bacteria, dermatophytes and yeasts. Dermatophytes were found to be the most susceptible microorganisms followed by yeasts, whereas bacteria were the least susceptible. The mean diameter of the inhibition zone determined by the disc diffusion screening method increased with increasing Plai oil concentration between 6.25 and 50 vol %. The minimum bactericidal concentration (MBC) determined by the broth macrodilution method ranged from 0.62 to 2.5 vol % for Plai oil and from 52 to 79 mg/mL for the 5 wt % Plai oil gel, whereas the minimum fungicidal concentration (MFC) ranged from 0.31 to 1.25 vol % for Plai oil and from 13.8 to 39.5 mg/mL for the 5% Plai oil gel. PMID- 17128431 TI - Pterocarpus santalinus: an In Vitro study on its anti-Helicobacter pylori effect. AB - The anti-H. pylori activity of Pterocarpus santalinus (PS), a traditional herb, has been assessed and compared with that of bismuth subcitrate, through in vitro studies employing rat gastric epithelial cell cultures and H. pylori isolates from gastric mucosal biopsy patients. The MIC of PS was found to be 20 microg/mL. H. pylori was co-cultivated with rat gastric epithelial cells in the presence/absence of PS at its MIC. A reduction in the activity of urease, a normal appearance of the epithelial cells on electron microscopic examination, a decrease in lipid peroxidation and lactate dehydrogenase suggests the possible anti-H. pylori activity of PS. PMID- 17128432 TI - Herbal modulation of drug bioavailability: enhancement of rifampicin levels in plasma by herbal products and a flavonoid glycoside derived from Cuminum cyminum. AB - The bioavailability of rifampicin (RIF) in a fixed dose combination (FDC) used for the treatment of tuberculosis remains an area of clinical concern and several pharmaceutical alternatives are being explored to overcome this problem. The present study presents a pharmacological approach in which the bioavailability of a drug may be modulated by utilizing the herb-drug synergism. The pharmacokinetic interaction of some herbal products and a pure molecule isolated from Cuminum cyminum with RIF is shown in this paper. An aqueous extract derived from cumin seeds produced a significant enhancement of RIF levels in rat plasma. This activity was found to be due to a flavonoid glycoside, 3',5-dihydroxyflavone 7-O beta-D-galacturonide 4'-O-beta-D-glucopyranoside (CC-I). CC-I enhanced the Cmax by 35% and AUC by 53% of RIF. The altered bioavailability profile of RIF could be attributed to a permeation enhancing effect of this glycoside. PMID- 17128433 TI - Antimicrobial activity of volatile components and various extracts of the red alga Jania rubens. AB - The methanol, dichloromethane, hexane, chloroform and volatile oil extracts of the red alga Jania rubens were tested in vitro for their antimicrobial activity (five Gram-positive, four Gram-negative bacteria and Candida albicans ATCC 10239). GC-MS analysis of the volatile components of J. rubens identified 40 compounds which constituted 77.53% of the total. The volatile components of J. rubens consisted of n-docosane (6.35%), n-eicosane (5.77%) and n-tetratriacontane (5.58%) as major components. The methanol and chloroform extracts (4 mg/disc) showed more potent antimicrobial activity than the hexane and dichloromethane extracts and the volatile oil of J. rubens. PMID- 17128434 TI - Vasorelaxant effect of stilbenes from rhizome extract of rhubarb (Rheum undulatum) on the contractility of rat aorta. AB - The vascular relaxant effect of the rhizome extract of Rheum undulatum was evaluated with isolated rat thoracic aorta preparations. The methanol extract of the rhizome induced a concentration-dependent relaxation of aortic preparations precontracted with 0.3 microm phenylephrine (EC50 value: 5.8 microg/mL). The activity-guided fractionation of the extract led to the isolation of seven hydroxystilbene components as active principles, i.e. piceatannol, resveratrol, desoxyrhapontigenin, rhapontigenin, piceid, rhaponticin and epsilon-viniferin. Of these, piceatannol, a tetrahydroxystilbene, exhibited the most potent vascular relaxant effect in rat aortic preparations (EC50 value 2.4 microm). The vasorelaxant effect of piceatannol on endothelium-intact aorta rings was diminished completely by the removal of functional endothelium or by pretreatment of the aortic tissues with N(G)-nitro-l-arginine methyl ester. These results suggest that piceatannol may be the major mediator responsible for the vasorelaxing properties of the rhizome extract of Rheum undulatum and the vasorelaxant effects of the piceatannol may be mediated via endothelium-dependent nitric oxide signaling pathway. PMID- 17128435 TI - Antidermatophytic triterpenoids from Syzygium jambos (L.) Alston (Myrtaceae). AB - Chemical investigation of the EtOAc extract of the stem bark of Syzygium jambos (L.) Alston (Myrtaceae) afforded a number of known triterpenes such as friedelin, beta-amyrin acetate, betulinic acid and lupeol. Friedelin was submitted to a Baeyer-Villiger oxidation, and was also reduced with LiAlH(4) to give the known friedelolactone and friedelanol, respectively. These compounds were identified by comparison of NMR spectral data with those from the literature. The EtOAc extract and the isolated compounds were tested for their antidermatophytic activity against three dermatophyte species: Microsporum audouinii, Trichophyton mentagrophytes and Trichophyton soudanense, commonly found in Cameroon. Betulinic acid and friedelolactone were the most active compounds, and the most sensitive fungi were Trichophyton soudanense and Trichophyton mentagrophytes. PMID- 17128436 TI - A review of the biological and potential therapeutic actions of Harpagophytum procumbens. AB - Harpagophytum procumbens (Hp), commonly known as Devil's Claw is a perennial plant which thrives in arid conditions. For centuries, it has been used as a traditional treatment for a variety of illnesses, including fevers, skin complaints, arthritis and diseases of the digestive tract as well as an appetite stimulant. Since its introduction to Europe in the early twentieth century, it has become a popular antiinflammatory and analgesic preparation amongst herbalists for supportive or adjuvant treatment of degenerative joint diseases, tendonitis, headache, backache and menstrual pain. The validity of Hp as an effective antiinflammatory and analgesic preparation, particularly in the relief of arthritic symptoms, has been investigated in numerous animal, clinical and in vitro studies. Although some contradictory evidence exists, the majority of animal studies appear to indicate Hp as an effective antiinflammatory and analgesic preparation in the treatment of acute and subacute inflammation. Clinical trials support Hp as a beneficial treatment for the alleviation of pain and improvement of mobility in a variety of musculoskeletal conditions. Analysis of the in vitro and ex vivo studies that currently exist, indicate that Hp has significant effects on numerous proinflammatory markers. However, the exact mechanism(s) by which Hp may reduce inflammation remain to be elucidated. PMID- 17128437 TI - Inhibitory effect of Panax notoginseng on nitric oxide synthase, cyclo-oxygenase 2 and neutrophil functions. AB - A water extract of Panax notoginseng Buck F.H. Chen. (Arialiaceae) root (PN) is being used as a therapeutic agent to stop haemorrhages and as a tonic to promote health in Korean and Chinese medicine. The pharmacokinetic profiles of PN have not been accurately investigated. The preliminary aim was to elucidate the pharmacokinetic features of PN. First, the prevention of neutrophil functions was assessed. PN inhibited neutrophil functions, including degranulation, superoxide generation and leukotriene B4 production, without any effect on 5-lipoxygenase activity. PN reduced nitric oxide (NO) and prostaglandin (PG)E2 production in mouse peritoneal macrophages stimulated with lipopolysaccharide (LPS) while no influence on the activity of inducible NO synthase (iNOS), cyclo-oxygenase-2 (COX 2) or cyclo-oxygenase-1 (COX-1) was observed. PN significantly reduced mouse paw oedema induced by carrageenan. The results indicate that PN exerts antiinflammatory effects related to the inhibition of neutrophil functions and NO and PGE2 production, which could be due to a decreased expression of iNOS and COX 2. PMID- 17128438 TI - Chemoprevention of Scutellaria bardata on human cancer cells and tumorigenesis in skin cancer. AB - Scutellaria barbata D. Don (Lamiaceae) (SB) is a perennial herb, which is natively distributed throughout Korea and southern China. This herb is known in traditional Chinese medicine as Ban-Zhi-Lian and in traditional Korean medicine as Banjiryun. SB has been used as an antiinflammatory and antitumor agent. The SB showed strong growth-inhibitory activity and cancer chemopreventive activity in assays representing three major stages of carcinogenesis. The SB was found to act as an antimutagen; it mediated antiinflammatory effects; inhibited cyclooxygenase and hydroperoxidase functions (antipromotion activity). In addition, SB inhibited the development of preneoplastic lesions in carcinogen-treated mouse mammary glands in culture and inhibited tumorigenesis in a mouse skin cancer model. On the other hand, an inhibitory effect of SB on the growth of gynecological cancer cell lines such as HeLa cell and human ovary cancer (HOC) was shown. When HOC cells were treated with SB, the expression of cyclooxygenase-2 was inhibited. These data suggest that SB merits investigation as a potential cancer chemopreventive agent in humans, especially in gynecological cancers. PMID- 17128441 TI - Prostate specific antigen (PSA) testing. PMID- 17128442 TI - Erectile dysfunction. PMID- 17128443 TI - Exercise. PMID- 17128444 TI - Diet. PMID- 17128445 TI - Correction: rationale for Standard LD.3.110. PMID- 17128446 TI - Parieto-frontal interactions, personal space, and defensive behavior. AB - In the monkey brain, two interconnected cortical areas have distinctive neuronal responses to visual, tactile, and auditory stimuli. These areas are the ventral intraparietal area (VIP) and a polysensory zone in the precentral gyrus (PZ). The multimodal neurons in these areas typically respond to objects touching, near, or looming toward the body surface. Electrical stimulation of these areas evokes defensive-like withdrawing or blocking movements. These areas have been suggested to participate in a range of functions including navigation by optic flow, attention to nearby space, and the processing of object location for the guidance of movement. We suggest that a major emphasis of these areas is the construction of a margin of safety around the body and the selection and coordination of defensive behavior. In this review, we summarize the physiological properties of these brain areas and discuss a range of behavioral phenomena that might be served by those neuronal properties, including the ducking and blocking reactions that follow startle, the flight zone of animals, the personal space of humans, the nearby, multimodal attentional space that has been studied in humans, the withdrawal reaction to looming visual stimuli, and the avoidance of obstacles during self-motion such as locomotion or reaching. PMID- 17128447 TI - Immune and neurological effects of work and environment. Proceedings of an international conference. April 21-24, 2002. Taiyuan, China. PMID- 17128448 TI - Controlling immune response may cut bird flu death rate. PMID- 17128449 TI - Raising awareness of cardiac tumours. PMID- 17128450 TI - Gene symbol: ABCA4. Disease: Stargardt disease 1. PMID- 17128451 TI - Gene symbol: F5. Disease: F5 type 1. PMID- 17128452 TI - Gene symbol: PANK2. Disease: pantothenate kinase-associated neurodegeneration (PKAN). PMID- 17128453 TI - Gene symbol: PANK2. Disease: pantothenate kinase-associated neurodegeneration (PKAN). PMID- 17128455 TI - Gene symbol: AR. Disease: androgen insensitivity syndrome. PMID- 17128456 TI - Gene symbol: AR. Disease: androgen insensitivity syndrome. PMID- 17128457 TI - Gene symbol: AR. Disease: androgen insensitivity syndrome. PMID- 17128458 TI - Gene symbol: PANK2. Disease: pantothenate kinase-associated neurodegeneration (PKAN). PMID- 17128459 TI - Gene symbol: NT5C3. Disease: pyrimidine 5'-nucleotidase (P5'N) deficiency. PMID- 17128460 TI - [Pneumology in the year 2005]. PMID- 17128461 TI - Gene symbol: F5. Disease: F5 type I. PMID- 17128462 TI - Gene symbol: JAG1. Disease: tetralogy of Fallot. PMID- 17128464 TI - Gene symbol: TYR. Disease: albinism, oculocutaneous 1. PMID- 17128465 TI - Gene symbol: msh2. Disease: hereditary nonpolyposis colorectal cancer. PMID- 17128466 TI - Gene symbol: NDP. Disease: Norrie disease. PMID- 17128467 TI - Gene symbol: MECP2. Disease: Rett syndrome. PMID- 17128468 TI - Gene symbol: MECP2. Disease: Rett syndrome. PMID- 17128469 TI - Gene symbol: ITGA2B. Disease: Glanzmann thrombasthenia. PMID- 17128470 TI - Gene symbol: F5. Disease: F5 type I. PMID- 17128472 TI - Gene symbol: F8. Disease: F8. PMID- 17128473 TI - Gene symbol: NOTCH3. Disease: CADASIL. PMID- 17128474 TI - Gene symbol: COL1A2. Disease: osteogenesis imperfecta type II. PMID- 17128475 TI - Gene symbol: PANK2. Disease: pantothenate kinase-associated neurodegeneration (PKAN). PMID- 17128476 TI - Gene symbol: PANK2. Disease: pantothenate kinase-associated neurodegeneration (PKAN). PMID- 17128480 TI - Gene symbol: PANK2. Disease: pantothenate kinase-associated neurodegeneration (PKAN). PMID- 17128481 TI - Gene symbol: PANK2. Disease: pantothenate kinase-associated neurodegeneration (PKAN). PMID- 17128482 TI - Gene symbol: SGSH. Disease: Sanfilippo type A syndrome, mucopolysaccharidosis IIIA. PMID- 17128483 TI - Gene symbol: PON1. Disease: coronary artery disease. PMID- 17128484 TI - Gene symbol: AR. Disease: androgen insensitivity syndrome. PMID- 17128485 TI - Gene symbol: AR. Disease: androgen insensitivity syndrome. PMID- 17128486 TI - Gene symbol: AR. Disease: androgen insensitivity syndrome. PMID- 17128487 TI - Gene symbol: AR. Disease: androgen insensitivity syndrome. PMID- 17128490 TI - Gene symbol: CRB1. Disease: early onset retinitis pigmentosa. PMID- 17128491 TI - Gene symbol: KCNQ1. Disease: LQT1. PMID- 17128492 TI - [Antihypertensive agents reduce the risk of apoplexy with 30-40%]. PMID- 17128493 TI - [A 24-year old woman with rash and swelling when using oral contraceptives and during pregnancy]. PMID- 17128494 TI - [Estrogen-dependent angioedema]. PMID- 17128496 TI - [Can the millennium goals be attained?]. PMID- 17128497 TI - EU re-opens debate on animal experimentation. PMID- 17128498 TI - Bibliography. Current World Literature. Cystic fibrosis. PMID- 17128499 TI - A salute to patient participation! PMID- 17128500 TI - Bibliography. Current world literature. Immunology. PMID- 17128501 TI - Plants, RNAi, and the Nobel Prize. PMID- 17128502 TI - Coronary heart disease in European South Asians. PMID- 17128503 TI - Protein-mediated energy-dissipating pathways in mitochondria. AB - Mitochondrial production of reactive oxygen species (ROS) is a well-established fact of fundamental importance to aging and etiology of many pathologies with serious public health implications. The ROS production is an innate property of mitochondrial biochemistry inseparable from the oxidative metabolism. Recent discoveries indicate that in addition to several ROS-detoxifying enzyme systems, which remove ROS, mitochondria may also be able to limit their ROS production by the mechanism comprising several protein-mediated energy-dissipating ("uncoupling") pathways. Although the physiological significance and in vivo modus operandi of these pathways remain to be elucidated, several proteins potentially capable of energy dissipation are known. This mini-review addresses the identity of mitochondrial protein-mediated energy-dissipating pathways and the experimental evidence to their role in controlling ROS production. PMID- 17128504 TI - Clinical challenges and images in GI. Yersinia enterocolitica mesenteric adenitis and terminal ileitis. PMID- 17128505 TI - Clinical challenges and images in GI. A gastric glomus tumor. PMID- 17128507 TI - Three quarters of AIDS deaths occur in sub-Saharan Africa. PMID- 17128509 TI - U.S. healthcare: "the dog's dinner". PMID- 17128513 TI - The OSMA. PMID- 17128514 TI - [Biodegradability and degradation mechanism of 3-fluorophenol by the activated sludge]. AB - An acclimated activated sludge was examined for its ability to degrade 3 fluorophenol in aerobic batch cultures. The result indicated that the organism degrades up to 100 mg/L 3-fluorophenol completely with approximately 100% fluoride anion release within 16 h. 3-Fluorophenol can serve as the sole carbon source and energy source for the organism. The acclimated activated sludge can degrade 3-fluorophenol effectively. The degradation mechanism study reveal that the initial step in the aerobic biodegradation of 3-fluorophenol is its transformation to fluorocatechol. Following transformation of the fluorophenol to fluorocatechol, ring cleavage by catechol 1, 2-dioxygenase proceedes via an ortho cleavage pathway, then defluorination occurres. PMID- 17128515 TI - JNK gives axons a second chance. PMID- 17128516 TI - In vivo chemical sensors: tackling biocompatibility. PMID- 17128517 TI - What does nanofluidics have to offer? PMID- 17128523 TI - Ropinirole: new indication. Restless legs: disproportionate adverse effects. AB - (1) The restless legs syndrome consists of unpleasant sensory and motor symptoms of varying intensity in the lower limbs. Symptoms occur at rest, seated or lying down, are more intense in the evening and at night, and are relieved by moving the limb. This syndrome does not cause serious physical complications. When sleep disturbances occur, non drug methods should be tried first. (2) Ropinirole is a dopaminergic agonist initially marketed for the treatment of Parkinson's disease. It is the first drug to be approved for restless legs syndrome in France. (3) Three double-blind randomised placebo-controlled trials with similar designs showed minimal differences on a composite rating scale. After 12 weeks of treatment, ropinirole led to an improvement of about 3 points on a 40-point scale compared with placebo. (4) A 12-week double-blind randomised controlled trial and including patients who had "responded" to ropinirole showed a lower relapse rate in the group that continued to use ropinirole (32.6%) instead of switching to placebo (57.8%). However, we do not know if this was because of continued drug efficacy or a rebound effect in the placebo group. (5) The adverse effects of ropinirole in patients with restless legs syndrome had already been observed in the treatment of Parkinson's disease, and included nausea, vomiting, drowsiness, a sudden urge to sleep, syncope, hypotension, and hallucinations. (6) An increase in the severity of restless legs symptoms, typically seen with levodopa, was not evaluated in clinical trials of ropinirole. Some cases have nevertheless been reported. They describe the appearance of symptoms increasingly early in the evening, then in the afternoon, or as a rebound effect in the morning or the latter part of the night. Their intensity increases and can affect other parts of the body. (7) In practice, ropinirole has a negative risk-benefit balance in restless legs syndrome, which is a minor health disorder. PMID- 17128524 TI - Treprostinil: new drug. Pulmonary artery hypertension: just another (disappointing) prostacycline analogue. AB - (1) Standard treatment for pulmonary artery hypertension usually combines a calcium channel blocker with an anticoagulant, supplemental oxygen, a digitalin and a diuretic, with only limited efficacy. When added to this standard treatment, long-term continuous intravenous epoprostenol (prostacycline) infusion improves survival time and quality of life in patients with severe primary pulmonary artery hypertension, but at a cost of many adverse effects, some of which can be serious. Bosentan, an endothelin receptor antagonist, is an oral alternative. (2) Following the approval of inhaled iloprost, another prostacycline analogue, treprostinil, has been approved for use as a continuous subcutaneous infusion. (3) Its clinical evaluation is based on 2 randomised double-blind placebo-controlled trials with disappointing clinical results: the 6 minute walking distance increased by only 10 meters after 12 weeks of treatment. This modest degree of improvement in an intermediate outcome is unlikely to translate into a tangible improvement in quality of life. (4) The only available comparison with epoprostenol is a clinical pharmacology study in about 20 patients, with no tangible clinical benefit. (5) The adverse effect profile of treprostinil is the same as that of epoprostenol, and is mainly due to its vasodilatory properties (diarrhoea, ankle swelling). In addition, pain and other local reactions are very frequent at the point of infusion. (6) Treprostinil must be administered as a continuous subcutaneous infusion; this is not convenient, but it is easier to set up than central intravenous epoprostenol infusion. PMID- 17128525 TI - Interferon gamma-1b: new indication. Severe malignant osteopetrosis: too many unknowns. AB - (1) Severe malignant osteopetrosis is a very rare disease. The principal manifestations are anaemia, infections, sensory disorders and fractures, due to generalised bone condensation. The disease is generally fatal in childhood. The only treatment capable of modifying the natural outcome is bone marrow transplantation. The benefits of high-dose steroids and calcitriol are usually modest and transient. (2) Severe malignant osteopetrosis is a new licensed indication for interferon gamma-1b, a drug known to reduce the incidence of severe infections in children with chronic septic granulomatosis. (3) An unblinded trial involving 15 children with a mean age of about one year compared calcitriol plus interferon gamma-1b with calcitriol alone. The time to treatment failure was longer with the combination, based on a combined endpoint chosen to make the statistical analysis more sensitive. (4) A clinical trial involving 15 patients, who were compared with a historical series of 94 untreated patients, provided ambiguous results. (5) In these trials the main adverse effect of interferon gamma-1b was a flu-like syndrome. (6) Given the gravity of severe malignant osteopetrosis, the limited available treatment options, and the rarity of serious adverse events with interferon gamma-1b, evaluation of this therapy should continue. PMID- 17128526 TI - Oseltamivir: cutaneous and neurological adverse effects in children. AB - (1) Oseltamivir is an antiviral drug used for influenza. It has only been tested in children to a limited extent. (2) A few deaths have been reported in children receiving oseltamivir, in some cases due to neurological causes. All of these deaths occurred in Japan. The European Medicines Agency has described 2 deaths, both attributed to suicide, in adolescents aged 14 and 17 who were taking oseltamivir. Both patients had developed behavioural disorders before their death. (3) According to the US Food and Drug Administration, neuropsychiatric disorders were reported in 32 children (mainly in Japan), and include delirium, behavioural disorders, hallucinations, convulsions and confusion. (4) Disturbing neurological toxicity has been observed in young rats exposed to oseltamivir, leading the American and European agencies to warn against the use of oseltamivir in infants less than one year old. (5) Severe cutaneous adverse effects, including cases of Lyell syndrome, were reported in children taking oseltamivir. (6) In practice, oseltamivir is only modestly effective in the prevention of influenza and treatment of suspected influenza. There is no evidence of an effect on complications of the flu in adults or children at higher risk. The possibility of serious adverse effects should be weighed against the limited benefit of oseltamivir. PMID- 17128527 TI - Thyroid extracts as slimming aids: danger! PMID- 17128528 TI - Pharmacological prevention of migraine: to be considered case by case. AB - (1) Migraines are characterized by recurrent headaches generally lasting between 4 and 72 hours and disappearing without complication. They can be incapacitating, owing to their frequency and/or intensity. (2) Many drugs have been used to prevent migraines. One of the most common outcome measures used in clinical trials is the proportion of responder patients, defined as those in whom the monthly frequency of migraines is at least halved. On average, about one-third of patients respond to placebo in clinical trials. (3) Propranolol is the betablocker with the best-documented efficacy: in absolute terms the response rate is about 30% higher than with placebo. The adverse effects of betablockers are mainly cardiovascular and neuropsychological. (4) Valproic acid, an anticonvulsant, is about as effective as propranolol, and its adverse effects are generally acceptable. (5) Amitriptyline is the antidepressant with the best documented preventive effects, with a response rate about 20% higher than placebo. Its principal adverse effects are due to its atropinic action. Amitriptyline can also have a sedative effect. (6) Flunarizine also has documented efficacy, but this "hidden neuroleptic" can cause extrapyramidal disorders and weight gain. (7) Among the serotonergic antagonists, methysergide has documented efficacy but long-term treatment can lead to serious retroperitoneal, pulmonary or cardiac fibrosis. Pizotifen causes drowsiness or weight gain in about 50% of patients. (8) The choice of preventive treatment for migraine must be based on the balance between efficacy (compared to placebo) and adverse effects. In practice, the first choice drug is propranolol. (9) Because the frequency of migraines fluctuates over time, withdrawal of prophylaxis should be attempted on a regular basis, with the patient's consent. PMID- 17128529 TI - Granulocytic growth factors and cancer-related neutropenia: limited effects. AB - (1) Cancer chemotherapy often causes haematological complications, in particular neutropenia, which can have grave consequences in terms of the risk of infections (increasing with the degree and duration of neutropenia) and the need for modifications in chemotherapy protocols (longer intervals between cycles or dose reductions). (2) In France, three haematopoietic growth factors are licensed to stimulate leukocyte production: filgrastim (unglycosylated granulocyte colony stimulating factor (G-CSF)), pegfilgrastim (pegylated filgrastim), and lenograstim (glycosylated G-CSF). (3) The main adverse effects of these three growth factors are joint and bone pain, a flu-like syndrome, and reactions at the injection site. (4) G-CSF has provided disappointing results in primary prevention, and its use is only justified for patients receiving chemotherapy that causes febrile neutropenia in at least 40% of cases: patients with acute leukaemia, elderly patients, and patients with cancer-related neutropenia or poor general status, etc. In these patients, G-CSF reduces the incidence of febrile neutropenia and, possibly, the risk of hospitalisation. A meta-analysis of 11 comparative trials involving about 1500 patients with non Hodgkin's lymphoma showed only a reduction in the incidence of febrile neutropenia and infections. (5) In the prevention of recurrences of neutropenia on continuing chemotherapy, only one trial, involving patients aged over 60 with high-grade non Hodgkin's lymphoma, showed an improvement in survival time with filgrastim (survival rate: 64.3% with filgrastim versus 49% with placebo, after a median follow-up of 40 months). Until these results are confirmed in other clinical trials, reduction in the intensity of chemotherapy (dosage, frequency) is generally recommended. (6) Curative treatment of neutropenia with G-CSF is only warranted for febrile patients with a high risk of severe infections requiring lengthy hospitalisation. Two meta-analyses, one including 8 trials and the other including 13 trials, involving a total of about 1500 patients, only showed a reduction in the length of hospitalisation. PMID- 17128530 TI - Screening mammography. PMID- 17128531 TI - [Subclinical hypothyroidism 2]. PMID- 17128532 TI - [Economic profit when injury treatment is transferred from emergency departments to general practice?]. PMID- 17128533 TI - [Role and limitation of internal medicine in therapy of patients with anorexia nervosa]. PMID- 17128534 TI - [Cognitive-behavioral therapy of behavior restriction for patients with anorexia nervosa]. PMID- 17128535 TI - Rethinking anticancer screening strategies saving lives at front line. Results from SESy_Europe task force. PMID- 17128536 TI - The surgicaI unsupersizing of America. PMID- 17128537 TI - Health care worker, vaccinate thyself: toward better compliance with influenza vaccination. PMID- 17128538 TI - Bariatric surgery: part of the answer to the obesity epidemic. PMID- 17128539 TI - The controversy over long-acting beta agonists: examining the evidence. AB - In the Salmeterol Multicenter Asthma Research Trial (SMART), patients receiving the long-acting beta agonist salmeterol--particularly African Americans--had a statistically significantly higher risk of fatal or potentially fatal asthma episodes. As a result, medications that contain salmeterol (Serevent, Advair) or formoterol (Foradil) carry a "black box warning." However, the benefits of these drugs continue to outweigh the risks, if they are used appropriately. PMID- 17128541 TI - Patient information: Bariatric surgery: is it right for you? PMID- 17128540 TI - Risks and benefits of bariatric surgery: current evidence. AB - Patients typically lose more than 50% of their excess weight after bariatric surgery. Obesity-related diseases markedly improve, reducing cardiovascular risk and improving life expectancy. Obese patients lose more weight with bariatric surgery than with medical weight-loss treatment. PMID- 17128542 TI - Improving influenza vaccination rates among adults. AB - Influenza remains an important cause of illness and death in this country. Even though we have safe and effective vaccines, vaccination rates among the elderly and other high-risk groups remain static and well below national goals. Health care providers can boost these vaccination rates by educating themselves, by recommending that their patients be vaccinated, and by implementing evidence based strategies such as programs to remind themselves and patients to be vaccinated, to utilize standing orders for nurses or other qualified professionals to offer and administer vaccines, and to provide feedback on performance. We should also consider alternative paradigms for vaccine delivery, and be sure to be vaccinated ourselves. PMID- 17128543 TI - Which agents should we use to treat and prevent influenza in 2006-2007? PMID- 17128544 TI - Endocrinology update 2006. AB - Endocrinology has recently witnessed several important developments: The Epidemiology of Diabetes Interventions and Complications study, a follow-up to the landmark Diabetes Control and Complications trial, found that strict glucose control early in the course of type 1 diabetes reduces the risk of microvascular and cardiovascular complications and provides prolonged benefits even if intensive control is not so tightly maintained. Inhaled insulin preparations are now available for mealtime coverage. We now have two new injectable medications for diabetes; pramlintide (Symlin) and exenatide (Byetta) are good adjuncts for patients with both type 1 and type 2 diabetes who have trouble reaching their hemoglobin A1c target, and they can help control and even reduce weight. Thyroxine (T4), instead of being merely a "prohormone," has been found to have direct actions on cells, leading to rapid clinical effects and possibly oncogenesis and angiogenesis. The therapeutic range for thyrotropin (TSH) may be much narrower than traditionally believed: some have proposed that the normal range should be redefined as 0.4 to 2.5 mIU/L. New evidence shows that vitamin D is important for more than calcium control and may help prevent type 1 diabetes. PMID- 17128545 TI - Does this patient have primary progressive aphasia? AB - Primary progressive aphasia (PPA) is a distinct clinical entity in which the patient develops language deficits while other cognitive domains remain relatively preserved until late in the course of the illness. The diagnosis can be relatively clear through an appropriate diagnostic approach based on the history and physical examination. There is no cure, but speech therapy is beneficial in this illness. PMID- 17128546 TI - A 48-year-old man with acute, 'knife-like' rectal pain. PMID- 17128547 TI - Pressure on your health benefits. PMID- 17128548 TI - The buzz on bees. PMID- 17128549 TI - How to give to the little guys. PMID- 17128550 TI - The celebrity of disease. PMID- 17128551 TI - Solving the SIDS mystery. PMID- 17128552 TI - To have and have not. PMID- 17128553 TI - IPPS FY'07 final rule contains 3-year phase-in for DRG cost weights, 3.4 percent payment increase. PMID- 17128554 TI - Changing of the guard. PMID- 17128555 TI - The uninsured: a growing crisis that demands action. PMID- 17128556 TI - Beyond the karyotype: are new screening methods needed for girls with Turner's syndrome? PMID- 17128557 TI - The treatment of Graves' disease in children. AB - Graves' disease is the most common form of hyperthyroidism in childhood. Current treatment options include antithyroid medications, surgery, and radioactive iodine. Medical therapy is generally associated with long-term remission rates of less than 25% and a small risk of serious adverse reactions that include hepatic failure and bone marrow suppression. Total thyroidectomy is associated with very high cure rates and a small risk of hypoparathyroidism and recurrent laryngeal nerve damage. When radioactive iodine is used at appropriate doses, there is a very high cure rate without increased risks of thyroid cancer or genetic damage. Clinicians caring for the child or adolescent with Graves' disease are thus faced with using medications with potential short-term and long-term toxicity, for a condition in which spontaneous remission occurs in the minority of pediatric patients. Definitive therapy in the form of surgery or radioactive iodine is necessary and unavoidable for the majority of pediatric patients with Graves' disease. PMID- 17128558 TI - Detection of hidden Y mosaicism in Turner's syndrome: importance in the prevention of gonadoblastoma. AB - The presence of Y chromosome fragments in patients with Turner's syndrome (TS) is known to increase the risk of gonadoblastoma. The investigation of Y sequences is usually performed only in the presence of marker chromosomes and therefore does not rule out the presence of hidden mosaicism in patients with 45,X TS without any marker. AIMS: 1. To investigate the presence of hidden Y mosaicism in non mosaic 45,X patients with TS, using samples from different tissues, and its association with the development of gonadoblastoma. STUDY DESIGN: Twenty patients with a 45,X karyotype were studied. The SRY and DYZ3 sequences were amplified by PCR, using genomic DNA from peripheral blood, oral epithelial cells and hair roots. Prophylactic gonadectomy was offered to the Y-positive patients. RESULTS: The analysis of the different tissues revealed that seven (35%) out of the 20 patients studied presented hidden chromosome Y mosaicism. Four of these patients underwent prophylactic gonadectomy, and bilateral gonadoblastoma was found in one of them. CONCLUSIONS: A systematic search for hidden Y chromosome mosaicism in patients with TS and 45,X karyotype is justified by the possibility of developing gonadoblastoma. PMID- 17128559 TI - Newborn screening levels of 17-hydroxyprogesterone in very low birth weight infants and the relationship to chronic lung disease. AB - OBJECTIVES: 17-Hydroxyprogesterone (17-OHP), an intermediary hormone in cortisol synthesis, has been shown to be elevated in premature infants. However, the relationship between levels of 17-OHP with chronic lung disease (CLD) have not been extensively explored. The objective of this study was to determine whether there is an association between CLD and levels of 17-OHP in a population of very low birth weight infants. STUDY DESIGN: Cohort study of very low birth weight infants cared for at a single level 3 NICU during a 3-year period from July 2001 July 2004, n=435. Infants had a minimum of one screen for 17-OHP. 17-OHP was measured on the 5th day of life and at 2-4 weeks of life as part of the State of Delaware Newborn Screening Program. Statistical analysis included chi-squared, Pearson correlation, and logistic regression. RESULTS: Levels of 17-OHP were higher at the time of the 1st screen compared to the 2nd screen (42.2 +/- 36.7 vs 23.5 +/- 32.3 ng/ml, respectively, p = 0.01). After controlling for potential confounding variables, gestational age and prenatal steroids were independently associated with 17-OHP. However, logistic regression analysis showed no association between a 1 log increase in levels of 17-OHP with the outcomes of CLD (odds ratio 1.7, 95% CI 0.7-3.8), or death and/or CLD (odds ratio 2.1, 95% CI 0.9 4.8). CONCLUSIONS: In our population of very low birth weight infants elevated levels of 17-OHP were not associated with the development of CLD. PMID- 17128560 TI - Anabolic steroid and gonadotropin releasing hormone analog combined treatment increased pubertal height gain and adult height in two children who entered puberty with short stature. AB - We studied the effect of gonadal suppression treatment in combination with anabolic steroid on pubertal height gain and adult height in two children who entered puberty with short stature. Patient 1 was a female with idiopathic short stature. She received combined treatment with an anabolic steroid (stanozolol) and a gonadotropin releasing hormone analog (leuprorelin acetate). Her pubertal height gain was 28.5 cm, which is greater than that in normal height girls (20-25 cm). Patient 2 was a male with Aarskog syndrome. Although his growth hormone (GH) secretion was normal, he received GH treatment. Since GH administration did not accelerate his growth, he received combined treatment with stanozolol and leuprorelin acetate. His pubertal height gain was 27.0 cm, which is greater than that observed in GH deficient boys treated with GH alone (21.9 cm). Combined treatment with stanozolol and leuprorelin acetate appears to be effective in increasing pubertal height gain and adult height in children who enter puberty with short stature. PMID- 17128561 TI - Relationships of IGF-I and andrrogens to skeletal maturation in obese children and adolescents. AB - IGF-I and androgens are postulated to accelerate skeletal maturation in obese children. METHODS: We studied weight status (BMI-SDS), height-SDS, IGF-I, cortisol, DHEA-S, and testosterone in 356 obese children (aged 4-15 years; 54% females) and correlated them to differences between bone age and chronological age (deltaBA-CA). Direct multivariate linear regression analyses were conducted for the dependent variable deltaBA-CA, including BMI, age, gender, pubertal stage, IGF-I-SDS, cortisol, DHEA-S, and testosterone as independent variables separately in prepubertal and pubertal girls, and prepubertal and pubertal boys. RESULTS: Height-SDS (r = 0.52), IGF-I (r = 0.33), and IGF-I-SDS (r = 0.36) were significantly (p < 0.001) correlated to deltaBA-CA. In multiple regression analyses, BMI and IGF-I-SDS were significantly positively (p < 0.001) correlated to deltaBA-CA independently of gender and pubertal stage. Testosterone was significantly positively correlated to detaBA-CA only in prepubertal girls (p = 0.035). CONCLUSIONS: Since IGF-I concentrations were positively associated to deltaBA-CA independently of pubertal stage and gender, we put forward the hypothesis that this hormone may contribute to acceleration of skeletal maturation in obese children. PMID- 17128562 TI - Predictors of metabolic control at one year in a population of pediatric patients with type 2 diabetes mellitus: a retrospective study. AB - BACKGROUND/OBJECTIVES: The rising prevalence of pediatric type 2 diabetes mellitus (DM2) and non-adherence to diabetes regimens pose challenges to obtaining optimal control. This study evaluated factors that may impact glycemic control (HbA1c): age, Tanner stage, body mass index (BMI), total daily insulin (TDD), metformin dose (MET), activity level, frequency of clinic visits and adherence. METHODS: One-year data from 72 patients (ages 8.6-17.8 years) were collected retrospectively. From that sample, 57 patients who continued to attend clinic for the entire year were assessed and divided into optimal and suboptimal HbA1c control groups. RESULTS: All factors measured were similar in the two groups, except for lower initial and 1.0-year HbA1c, TDD, and rates of missing MET and insulin in the optimal HbA1c control group. CONCLUSIONS: Initial glycemic status and adherence rate predicted metabolic control at one year. Early identification of DM2 may improve metabolic outcome, which may improve medical regimen adherence. PMID- 17128563 TI - Serum lipid profile in children receiving anti-epileptic drug monotherapy: is it atherogenic? AB - The effect of anti-epileptic drugs (AEDs) on serum lipid profile is controversial in children as well as in adults. We longitudinally studied serum lipid profile in 34 newly diagnosed epileptic children receiving AED monotherapy with valproic acid (VPA), carbamazepine (CBZ) or phenobarbital (PB). Serum levels of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triglycerides (TGs), apolipoprotein Al (Apo A1) and apolipoprotein B (Apo B) were measured at baseline and after 2 years of AED monotherapy. Atherosclerotic indices of TC/ HDL-C and Apo A1/Apo B ratios were calculated. Although there were some alterations in serum lipid profile with AED without statistical significance, the atherosclerotic indices of TC/HDL-C and Apo A1/Apo B ratios did not change significantly after 2 years of monotherapy. Only serum TGs levels significantly decreased with VPA monotherapy. These data suggest that 2 years AED monotherapy with VPA, CBZ or PB did not cause a significant level of concern for an atherogenic effect in children with epilepsy. PMID- 17128564 TI - Compound heterozygosity of a frameshift mutation in the coding region and a single base substitution in the promoter of the ACTH receptor gene in a family with isolated glucocorticoid deficiency. AB - Isolated glucocorticoid deficiency (IGD) is an autosomal recessive syndrome characterized by glucocorticoid insufficiency without mineralocorticoid deficiency. Mutations in the coding region of the ACTH receptor (MC2R) have been reported in several families with IGD. We amplified and sequenced the entire MC2R coding region in a new family with IGD. The proband was found to be heterozygous (paternal allele) for the mutation Gly217fs, which changes the open reading frame of the MC2R protein resulting in a truncated receptor. No other abnormality was found in the MC2R coding region. However, sequencing of the promoter region of the MC2R gene (-1017/44 bp) of the proband revealed a heterozygous T-->C substitution in the maternal allele at -2 bp position from initiation of the transcription start site. This substitution was found in only 6.5% in a healthy unrelated population. Constructs containing this polymorphism consistently showed a significant 15% decrease in promoter activity compared to wild type. In conclusion, we provide evidence that the IGD in this previously unreported family with ACTH resistance appears to be secondary to compound heterozygosity of a coding region and a promoter mutation in the MC2R gene. PMID- 17128565 TI - Novel compound heterozygous mutation of the MC2R gene in a patient with familial glucocorticoid deficiency. AB - Familial glucocorticoid deficiency (FGD) is an autosomal recessive disorder characterised by glucocorticoid insufficiency without mineralocorticoid deficiency. Here, we report a 2 year-old girl with FGD, showing tall stature and skin pigmentation, but no abnormalities of the external genitalia. Serum sodium, potassium and chloride levels were within normal ranges. Endocrinological analysis revealed low serum cortisol (<5.5 nmol/1), elevated plasma ACTH (875.2 pmol/1) and low 17alpha-hydroxyprogesterone (< 0.303 nmol/l). We suspected the patient of having FGD type 1. Direct and allele-specific sequence analyses of the melanocortin 2 receptor gene (MC2R) revealed compound heterozygous mutations (C21Y and R146H) in the MC2R gene. Her father and mother each had heterozygous C21Y and R146H mutations, respectively, without symptoms of glucocorticoid deficiency. This is the first report of FGD associated with a compound heterozygous mutation of C21Y and R146H in the MC2R gene. PMID- 17128566 TI - Complex urogenital malformation associated with female pseudohermaphroditism: caudal dysgenesis syndrome. AB - Caudal dysgenesis syndrome is a rare cause of female pseudohermaphroditism. This syndrome consists of absent perineal and anal openings in association with ambiguous genitalia, urogenital, colonic, and lumbosacral anomalies. We report a case of caudal dysgenesis syndrome in an infant who had non-palpable testes, bifid scrotum, a phallus-like structure and urethral atresia. Radiological evaluation revealed bilateral hydronephrosis, bifid uterus, cervix and vagina. Caudal dysgenesis syndrome should be considered in any female infant presenting with bilateral streak ovaries, and Mullerian and genito-urinary anomalies. PMID- 17128567 TI - Importance of thyroglobulin levels for diagnosis and monitoring of follicular thyroid carcinoma in an adolescent with severe iodine deficiency. AB - Optimal management of differentiated thyroid cancer in childhood is undetermined. During monitoring of thyroid carcinoma, serum thyroglobulin (hTG) levels provide valuable information. hTG levels not only increase in differentiated thyroid cancers but also in iodine deficiency because of compensation by the thyroid gland. A 14.6 year-old girl was diagnosed with nodular goiter, subclinical hypothyroidism and severe iodine deficiency. She had a very high hTG level. Despite benign fine-needle aspiration biopsy (FNAB), because the hTG level was still very high after treatment with LT4, thyroidectomy was undergone. Cytopathological examination showed minimally invasive follicular thyroid carcinoma. During follow-up, to exclude the presence of persistent/recurrent disease, the hTG level rose to an undesirably high level after withdrawal of TSH suppressive therapy, and radioiodine ablation therapy was applied. This report shows that even if there is an explanation for nodular goiter and high hTG levels, such as iodine deficiency, malignancy cannot be ruled out without thyroidectomy. FNAB is not reliable especially in iodine deficient areas. Serum hTG measurement is a valuable tool for both diagnosis and follow-up of differentiated thyroid carcinoma in children. PMID- 17128568 TI - Two years of growth hormone treatment in the first growth hormone deficient patient with cerebrofaciothoracic dysplasia. AB - We recently reported two siblings, a sister and a brother, with intrauterine growth retardation, microcephaly, short stature, mental retardation, facial dysmorphism and multiple costovertebral malformations. These features fit most with the diagnosis of cerebrofaciothoracic dysplasia, or Pascual-Castroviejo syndrome. The second sibling, our index patient, presented also with cleft palate and growth hormone (GH) deficiency, suggesting that endocrinological assessment should be performed in short patients with this syndrome, especially if midline defects are present. We present the results of 2 years GH treatment of this first GH deficient patient with cerebrofaciothoracic syndrome and compare the results to those observed in other genetic syndromes with GH deficiency. PMID- 17128569 TI - Minister puts onus on trusts to negotiate unbundled tariff. PMID- 17128570 TI - PCTs told to implement radical changes to LAAs. PMID- 17128571 TI - Latest IT delays a 'catastrophe'. PMID- 17128572 TI - Scotland motors into the future as England gears up for change. PMID- 17128573 TI - On clinical engagement. PMID- 17128574 TI - Managing a merger? Don't lose the plot. PMID- 17128575 TI - Barometer. PCTS Oct. 2006. PMID- 17128576 TI - Long-term conditions. Predicting the future. AB - The predictive risk project aims to reduce hospital bed use associated with long term conditions by targeting frequently admitted patients in the community. The project also uses an algorithm to identify patients at risk of future admissions, with evidence showing promising results to date. A different approach has been developed in Norfolk, where a coaching system aims to prevent admissions by encouraging at-risk patients to manage their own care. PMID- 17128577 TI - Living with lung cancer. PMID- 17128578 TI - Casting votes on healthcare. Midterm election referendums bring big money, potential impact on healthcare-mostly with anti-smoking proposals. AB - While there are plenty of healthcare-related issues on this year's midterm ballots, Joy Johnson Wilson, left, of the National Conference of State Legislatures, says there were many more that failed to make it. "The cost has gone up to get something on the ballot, and you need an organization to push those measures," she says. PMID- 17128579 TI - Hospitals' profitable year. AHA: '05 delivered aggregate profit margin of 5.3%. PMID- 17128580 TI - Two down, two up. Community, LifePoint join HCA in bad debt woes. PMID- 17128581 TI - HCA battles legal obstacles. PMID- 17128582 TI - MGMA talks payments, safety. PMID- 17128583 TI - System: no conflict for exec. Health Alliance comfortable with CEO's involvement. PMID- 17128584 TI - These are our rules. An updated code of ethics for Modern Nealthcare's editorial staff. PMID- 17128585 TI - On the comeback trail. Healthcare is an issue on the verge of making big waves in politics again. PMID- 17128586 TI - Many happy returns. Insurers flock back to the Medicare managed-care market, but how long will they stick around this time? PMID- 17128587 TI - Special effects. With moratorium over, specialty projects growing--slowly for now. PMID- 17128588 TI - CMS dangling carrot. Up to 50,000 dollars available in P4P plan. PMID- 17128589 TI - [New systems for colonic drug targeting]. AB - Colonic drug delivery systems are useful for the treatment of inflammatory bowel diseases (ulcerative colitis, Crohn's disease), some carcinomas, gastrointestinal infections and diseases that are sensitive to circadian rhythms, such as asthma, angina pectoris and arthritis, where an intention time delay in the absorption of the drug is required. The colon region is a site for the entry of peptides, proteins, nucleotides and vaccines into the systemic circulation due to local absence of digestive enzymes. Colonic drug targeting ensures direct treatment at the disease site and a possible reduction in the administered dose. A drug is not absorbed into the systemic circulation, so the associated systemic adverse effects are reduced. Colonic drug delivery systems are based on exploitation of characteristics that are unique to the gastrointestinal tract (GIT), such as pH gradient along the GIT, long transit time through the GIT, increased intraluminal pressure and a presence of bacterial microflora in these regions. PMID- 17128590 TI - [Some possibilities of using spectrometry in the near infrared region in drug quality control]. AB - Spectrometry in the near infrared region ranks among the modern methods of analytical chemistry and is of use also in drug quality control. The paper draws attention to some possibilities of using the method in this respect, including its possible use in the check-out of the identity of the content of closed packages of medicinal and auxiliary substances supplied to pharmacies. PMID- 17128591 TI - [Radionuclide X-ray fluorescence as an alternative method to pharmacopeial tests for heavy metals]. AB - Pharmacopeial tests for heavy metals require comparisons between test solution's colour after addition of thioacetamide and a control solution containing a known quantity of Pb. As this test neither informs about the type of heavy metals, nor about the concentrations of each of them, there is a reason for more frequent employment of instrumental methods such as AAS and AES, as well as ICP, NAA, and X-ray fluorescence. These methods provide us with quick and exact elemental analyses of impurities, thus becoming more and more important in the quality control of drugs and medicinal plants. PMID- 17128592 TI - [Plant metabolites as nootropics and cognitives]. AB - Nowadays several millions of people suffer from Alzheimer's disease and other types of dementia. Etiology of these diseases is not known very well. There occur different levels of neurotransmitters, the level of acetylcholine in the brain is decreased and pathological changes affect the brain tissue. Organic and toxic damage of the brain, free radicals, and other changes participate in the development of these diseases. Drugs as nootropics, cognitives, and neuroprotectives are commonly used to treat these diseases. Some of these drugs have often side and undesirable effects. In recent years some natural substances (galanthamine, huperzine A, vinpocetine), and standardized plant extracts (Ginkgo biloba L., Centella asiatica L.) Urban, Bacopa monniera L., Evolvulus alsinoides L.) are often used. These plant preparations produce fewer undesirable effects and the same effectiveness as the classic therapy, or these preparations are used as a supplement to the classic therapy. PMID- 17128593 TI - [A comparison of inhalatory beta2-mimetics with long-term action (formoterol vs. salmeterol)]. AB - Asthma is a serious global health problem. People of all age groups in all countries of the world suffer from this chronic disease of the respiratory tract, which may have a very serious and sometimes fatal course. Most beta2-mimetics with a rapid onset of effect act for 4 to 6 hours. Inhalatory beta2-mimetics with long-term action (LABA), formoterol and salmeterol, possess a period of action longer than 12 hours. Formoterol and salmeterol represent great progress in asthma management, particularly in combination with inhalatory corticosteroids (IKS). Their effective bronchodilatory properties and long-term improvement of pulmonary functions are a great clinical contribution for the patients. Both formoterol and salmeterol are strong and effective beta2-agonists, but their different chemical structures produce different pharmacological properties. Due to the fact that the onset of the effect of salmeterol is slower, it should not be used to treat acute symptoms or quickly deteriorating asthma. Formoterol exerts a rapid onset of the effect and high internal activity, thanks to which it can be used for relieving treatment. The present paper aims to characterize and compare the properties of long-term acting beta2-mimetics, with detailed focus on their two representatives, formoterol and salmeterol. PMID- 17128594 TI - [Studies of the properties of tablets made from directly compressible starch and its mixtures with directly compressible lactose]. AB - The paper studies the strength and disintegration time of compacts made from directly compressible Starch 1500 and its mixtures with directly compressible lactose Pharmatosa DCL 15 in different relative proportions in dependence on the added lubricant magnesium stearate and the model active ingredient ascorbic acid. The dry binders under study differ in their mechanisms of compression and thus in their sensitivity to the addition of the lubricant, which is manifested in decreased strengths of the compacts as well as the mechanism of disintegration of the compacts. The mixtures of substances under study were in the relative representations of 1:3, 1:1 and 3:1. The employed concentration of the lubricant magnesium stearate was 0.4%, that of ascorbic acid, 50%. The study has confirmed and quantitatively evaluated the sensitivity of directly compressible starch to the addition of magnesium stearate. A higher share of directly compressible lactose in the mixture decreased the sensitivity of tableting material to the addition of the lubricant and shortened disintegration time. The presence of stearate did not negatively influence disintegration time, with an exception of the mixture of Starch 1500 and Pharmatosa DCL 15 1:3. An addition of the model active ingredient ascorbic acid decreased the strength of the compacts and shortened the disintegration time in all tableting materials tested. PMID- 17128595 TI - [Flavonoids--main constituents of the leaves of Holodiscus discolor (Pursh) Maxim]. AB - The paper deals with the isolation and identification of constituents of the leaves of Holodiscus discolor (Pursh) Maxim. (Rosaceae). Three flavonoid glycosides of flavonol type were isolated from the methanolic extract: kaempferol 3-O-rhamnoside, quercetin-3-O-glucoside (isoquercitrin) and quercetin-3-O rhamnoside (quercitrin). Isolates were identified by physical-chemical data, by comparison with authentic samples and literature data. The above-mentioned compounds were isolated from Holodiscus discolor (Pursh) Maxim. for the first time. PMID- 17128596 TI - Illusions of competence during study can be remedied by manipulations that enhance learners' sensitivity to retrieval conditions at test. AB - Monitoring one's knowledge during study is susceptible to a foresight bias (Koriat and Bjork, 2005). Judgments of learning (JOLs) are inflated whenever information that is present at study and absent, but solicited, at test, such as the targets in cue-target paired associates, highlights aspects of cues that are less apparent when those cues are presented alone. The present findings demonstrate that foresight bias can be alleviated by study-test experience (Experiment 1), particularly test experience (Experiments 2 and 3), and by delaying JOLs after study (Experiment 4) and that both foresight bias and its alleviation have behavioral consequences, as measured by study time allocation (Experiment 5). Collectively, the findings suggest that overconfidence and misallocation of study time arise from a mismatch that is inherent to education- that the answer is present at study and absent at test--and that alleviating the problem requires creating conditions at study that sensitize learners to retrieval conditions at test. PMID- 17128597 TI - Word concreteness and encoding effects on context-dependent discrimination. AB - In three experiments, we investigated the effects of word concreteness and encoding instructions on context-dependent discrimination in verbal contexts, using Murnane, Phelps, and Malmberg's (1999) ICE (item, context, ensemble) theory as the framework Word concreteness was manipulated within participants, and encoding was manipulated between participants. It was hypothesized that the magnitude of context-dependent discrimination would be affected by both concreteness and encoding instructions. Imagery instructions resulted in context dependent discrimination for both concrete and abstract word pairs across all the experiments. Context-dependent discrimination was observed under rote instructions for concrete and abstract words, particularly when the same context word was paired multiple times with the targets. The results indicated that context-dependent discrimination is not dependent solely on the use of interactive imagery instructions or on word concreteness. PMID- 17128598 TI - Exploring the role of attention during memory retrieval: effects of semantic encoding and divided attention. AB - Research on attention and memory suggests that semantic encoding leads to retrieval that is highly susceptible to divided attention. Three experiments tested this proposition and showed that dividing attention did not selectively affect semantically encoded items. Participants encoded a list of words in one of two ways: semantically or phonetically. Later, memory was assessed using either a standard recognition test (Experiment 1) or a rhyme recognition test (Experiments 2 and 3). The participants took the memory test either alone (full attention) or while simultaneously performing a secondary task (divided attention). Recognition accuracy was reduced by divided attention on both recognition tests, and semantically and phonetically encoded words were equally affected. PMID- 17128599 TI - Determinants of lexical access in speech production: role of word frequency and age of acquisition. AB - The main determinants of lexical access in speech are considered to be a word's age of acquisition (AoA) and its frequency of occurrence in a speaker's experience. It is unclear whether and how these variables interact, although they are commonly observed to be correlated, for the few studies that address the issue have reported inconsistent findings. An influential view of AoA in lexical processing (Ellis and Lambon Ralph, 2000) predicts stronger frequency effects for items acquired later in life than for those acquired at an early age. Five experiments were designed to investigate the possible interaction of AoA and frequency effects in speech. We found that the interaction between word frequency and AoA was not robust and that, contrary to expectation, the effect of word frequency was greater for words acquired earlier in life than for those acquired later. The implications of our findings are discussed. PMID- 17128600 TI - What can subjective forgetting tell us about memory for childhood trauma? AB - In the present study, we examined the prevalence and predictors of subjective forgetting (i.e., self-reported amnesia) of child sexual abuse (CSA). Adults who, as children, were involved as victims in legal prosecutions were questioned about their CSA experiences, which had been documented in the 1980s, and about lost and recovered memory of those experiences. Males and individuals who experienced more severe abuse were more likely to report forgetting. The majority of individuals attributed their forgetting to active attempts to avoid thinking about the abuse. In contrast, when predictors of subjective forgetting were used to predict objective memory of abuse, more severe abuse and more extended legal involvement were associated with fewer memory errors. The differences between subjective and objective memory underscore the risks of using subjective measures to assess lost memory of abuse. PMID- 17128601 TI - The role of test structure in creating false memories. AB - In the Deese/Roediger-McDermott (DRM) paradigm, studying lists of semantic associates results in high rates of false recognition of a nonpresented critical word. The present set of experiments was designed to measure the contribution of additional processing of list items at test to this false memory effect. The participants studied sets of lists and then performed a recognition task for each set. In three experiments, using this paradigm, we investigated false recognition when the number of studied list items presented at test (0, 6, or 12) was manipulated. In Experiments 2 and 3, false recognition of critical lures associated to both studied and nonstudied lists increased significantly as the number of list items included in the test increased. These results indicate that processes occurring at retrieval contribute to false memory effects found with the DRM paradigm. PMID- 17128602 TI - Task interference from prospective memories covaries with contextual associations of fulfilling them. AB - One of the current issues in the field of prospective memory concerns whether having an intention produces a cost to other ongoing activities (called task interference). The evidence to date suggests that certain intentions held over the shorter term do interfere with other tasks. Because the cumulative effect of such costs would be prohibitively expensive in everyday life, the present study examined one means by which that interference may be reduced. Participants who formed a specific association to fulfilling an intention in a future context did not exhibit task interference over the intervening period until that context was encountered. This outcome was observed with both an event-based and a time-based prospective memory task. The results suggest that associating intention fulfillment with a specific context can eliminate task interference, and they emphasize the importance of studying intentions that are linked to future contexts versus those that are not. PMID- 17128603 TI - Do distractors interfere with memory for study pairs in associative recognition? AB - In an associative recognition task, distractors generally consist of a rearrangement of the items composing the study pairs. This makes it possible that processing the distractors generates retroactive interference on memory for the study pairs. In Experiment 1, we explored this possibility in a yes/no recognition test concerning previously learned arbitrary associations between visual symbols and auditory syllables. Rearranged pairs had a deleterious impact on the accuracy and the speed of responses to related correct pairs. This effect did not vary as a function of the number of training blocks, and furthermore, in Experiment 2, the same effect was observed for overlearned small multiplication facts. These results suggest that exposure to potentially confounding information generates interference even if this information is known to be incorrect. Some implications are outlined, especially with regard to the widespread use of multiple-choice tests in knowledge evaluation. PMID- 17128604 TI - Artificially induced valence of distractor words increases the effects of irrelevant speech on serial recall. AB - In a game context, nonwords either were artificially associated with negative valence or were in some sense neutral or irrelevant. Subsequently, participants memorized target words in silence or while attempting to ignore the negatively valent, irrelevant, or neutral auditory distractor nonwords. The presence of distractor nonwords impaired recall performance, but negative distractor nonwords caused more disruption than neutral and irrelevant distractors, which did not differ in how much disruption they caused. These findings conceptually replicate earlier results showing disruption due to valence with natural language words and extend them by demonstrating that auditory features that may possibly be confounded with valence in natural language words cannot be the cause of the observed disruption. Explanations of the irrelevant speech effect within working memory models that specify an explicit role of attention in the maintenance of information for immediate serial recall can explain this pattern of results, whereas structural models of working memory cannot. PMID- 17128605 TI - Proactive interference and cuing effects in short-term cued recall: does foil context matter? AB - Tehan and Humphreys's (1995, 1996) short-term cued recall paradigm showed that recall in short-term memory is cue driven. In critical trials, the participants studied two blocks of four words each and were required to forget the first block while remembering the second block. A foil in the first block (e.g., orange) was related to a target (e.g., carrot) in the second block. Proactive interference (PI) was evident when a retrieval cue was used that subsumed the foil and the target (e.g., type of juice), but not when a cue was used that subsumed only the target (e.g., type of vegetable). Four experiments were performed to examine the extent to which contextual organization in the foil block would enhance or diminish the foil's efficacy in creating PI. A novel condition was included in which the words in the foil block were studied in a phonologically related context but the target was cued semantically, and vice versa with a semantic context and phonological cue. There were no differences in recall accuracy between conditions with and without contextual organization, but reliable increases in foil intrusions were observed when contextual organization was present. Contextual organization enhanced the foil, rather than diminished it, but the strengthened foil generated PI only when the cue subsumed the foil and the target and had no effect when the cue subsumed only the target. The results are consistent with a cue-driven retrieval interpretation of short-term recall. PMID- 17128606 TI - Auditory and visual spatial working memory. AB - A series of experiments compared short-term memory for object locations in the auditory and visual modalities. The stimulus materials consisted of sounds and pictures presented at different locations in space. Items were presented in pure- or mixed-modality lists of increasing length. At test, participants responded to renewed presentation of the objects by indicating their original position. If two independent modality-specific and resource-limited short-term memories support the remembering of locations, memory performance should be higher in the mixed modality than in the pure-modality condition. Yet, memory performance was the same for items in both types of list. In addition, responses to the memory load manipulation in both modalities showed very similar declines in performance. The results are interpreted in terms of object files binding object and location information in episodic working memory, independently of the input modality. PMID- 17128607 TI - Associative priming in faces: semantic relatedness or simple co-occurrence? AB - In two experiments, we explored the effects of co-occurrence and semantic relationships in the associative priming of faces. In Experiment 1, pairs of computer-generated human faces were presented simultaneously (i.e., they co occurred) with no associated semantic information attached to them. A significant facilitation effect in the subsequent recognition of these paired faces (priming) was observed. Thus, repeatedly presenting faces together while keeping semantic information to a minimum appears to be enough to produce associative priming. In Experiment 2, the computer-generated faces were associated with semantic information and again presented ipairs. Priming effects arising from co occurrence and semantic relatedness were observed. The results from these experiments show that semantic relatedness is not the sole cause of the association between faces; co-occurrence plays a crucial role too. This conclusion has significant implications for the current computational models of face processing. PMID- 17128608 TI - Theory-based categorization under speeded conditions. AB - It is widely accepted that similarity influences rapid categorization, whereas theories can influence only more leisurely category judgments. In contrast, we argue that it is not the type of knowledge used that determines categorization speed, but rather the complexity of the categorization processes. In two experiments, participants learned four categories of items, each consisting of three causally related features. Participants gave more weight to cause features than to effect features, even under speeded response conditions. Furthermore, the time required to make judgments was equivalent, regardless of whether participants were using causal knowledge or base-rate information. We argue that both causal knowledge and base-rate information, once precompiled during learning, can be used at roughly the same speeds during categorization, thus demonstrating an important parallel between these two types of knowledge. PMID- 17128609 TI - The effect of emotion on interpretation and logic in a conditional reasoning task. AB - The effect of emotional content on logical reasoning is explored in three experiments. Theparticipants completed a conditional reasoning task (If p, then q) with emotional and neutral contents. In Experiment 1, existing emotional and neutral words were used. The emotional value of initially neutral words was experimentally manipulated in Experiments 1B and 2, using classical conditioning. In all experiments, participants were less likely to provide normatively correct answers when reasoning about emotional stimuli, compared with neutral stimuli. This was true for both negative (Experiments 1B and 2) and positive contents (Experiment 2). The participants' interpretations of the conditional statements were also measured (perceived sufficiency, necessity, causality, and plausibility). The results showed the expected relationship between interpretation and reasoning. However, emotion did not affect interpretation. Emotional and neutral conditional statements were interpreted similarly. The results are discussed in light of current models of emotion and reasoning. PMID- 17128610 TI - The role of perceptually represented structure in analogical problem solving. AB - Current models of analogical reasoning assume that representations of source examples and target problems occur in an amodal format--that is, a representation whose construction and processing are independent of activity in the perceptual and motor cortices of the brain. We examined the possible use of kinesthetic information--perceptual structures associated with the sensation of space and force--in the representation of source examples and target problems. Participants who recreated a source story while acting out the key elements were more likely to access the story when later working on the target problem than were participants who only verbally recreated the story or who verbally recreated it as well as sketched it. We argue that enactment made kinesthetic and spatial features more salient in participants' source story representations and that this aided performance. These results suggest that current models of analogical reasoning might be improved by including perceptual information as part of their representational schemes. PMID- 17128611 TI - People want to see information that will help them make valid inferences in human causal learning. AB - According to higher order reasoning accounts of human causal learning (e.g., Lovibond, Been, Mitchell, Bouton, and Frohardt, 2003; Waldmann and Walker, 2005) ceiling effects in forward blocking (i.e., smaller blocking effects when the outcome occurs with a maximal intensity on A+ and AX+ trials) are due to the fact that people are uncertain about the causal status of a blocked cue X in a forward blocking design when the outcome is always fully present on A+ and AX+ trials. This should not be the case for a reduced overshadowing cue Y (B- trials followed by BY+ trials). We tested this hypothesis by asking participants which additional information they preferred to see after seeing all learning trials. Results showed (1) that all participants preferred to see the effect of the blocked cue X over seeing the effect of the reduced overshadowing cue Y (Experiment 1), and (2) that more participants preferred to see the blocked cue X on its own when the outcome on A+ and AX+ trials was fully present than when the outcome on those trials had a submaximalintensity (Experiment 2). PMID- 17128612 TI - Learning errors from fiction: difficulties in reducing reliance on fictional stories. AB - Readers rely on fiction as a source of information, even when fiction contradicts relatively well-known facts about the world (Marsh, Meade, and Roediger, 2003). Of interest was whether readers could monitor fiction for errors, in order to reduce suggestibility. In Experiment 1, warnings about errors in fiction did not reduce students' reliance on stories. In Experiment 2, all subjects were warned before reading stories written at 6th- or 12th-grade reading levels. Even though 6th-grade stories freed resources for monitoring, suggestibility was not reduced. In Experiment 3, suggestibility was reduced but not eliminated when subjects pressed a key each time they detected an error during story reading. Readers do not appear to spontaneously monitor fiction for its veracity, but can do so if reminded on a trial-by-trial basis. PMID- 17128613 TI - Walking through doorways causes forgetting: situation models and experienced space. AB - We investigated the ability of people to retrieve information about objects as they moved through rooms in a virtual space. People were probed withobject names that were either associated withthe person (i.e., carried) or dissociated from the person (i.e., just set down). Also, people either did or did not shift spatial regions (i.e., go to a new room). Information about objects was less accessible when the objects were dissociated from the person. Furthermore, information about an object was also less available when there was a spatial shift. However, the spatial shift had a larger effect on memory for the currently associated object. These data are interpreted as being more supportive of a situation model explanation, following on work using narratives and film. Simpler memory-based accounts that do not take into account the context in which a person is embedded cannot adequately account for the results. PMID- 17128614 TI - Reading strategies and prior knowledge in learning from hypertext. AB - In two experiments, we identified two main strategies followed by hypertext readers in selecting their reading orders. The first consisted in selecting the text semantically related to the previously read section (coherence strategy), and the second in choosing the most interesting text, delaying reading of less interesting sections (interest strategy). Comprehension data revealed that these strategies affected learning differently as a function of the reader's prior knowledge. For low-knowledge readers, the coherence strategy supported better learning of the content. This effect seems to rely on the improvement of reading order coherence induced by this strategy. By contrast, for intermediate-knowledge readers the coherence and the interest strategies benefited comprehension equally. In both cases, learning was supported through the active processing induced by these strategies. Discussion focuses on resolving inconsistencies in the literature concerning whether or not hypertext supports better comprehension than does traditional linear texts. PMID- 17128615 TI - Are age-of-acquisition effects on object naming due simply to differences in object recognition? Comments on levelt (2002). AB - Levelt (2002) argued that apparent effects of word frequency and age of acquisition (AoA) reported in recent picture naming studies might actually be confounded effects operating at the level of object recognition, rather than relevant to theories of lexical retrieval. In order to investigate this issue, AoA effects were examined in an object recognition memory task (Experiments 1 and 2) and a word-picture verification task (Experiment 3) and compared with those found in naming tasks using the same pictures. Contrary to Levelt's concerns, the results of the three experiments show that the AoA effect on picture naming has a lexical origin and does not simply result from a possible confound of object identification times. PMID- 17128616 TI - Word order variation in spatial descriptions with adverbs. AB - Previous research has shown that in a three-term spatial reasoning task, the second premise of a German premise pair is especially easy to comprehend if (1) the prepositional object rather than the grammatical subject denotes the given entity, and if (2) the term denoting the given entity precedes the term denoting the new entity. Accordingly, the second premise is easiest to comprehend with noncanonical word order--that is, with the prepositional object in preverbal position denoting the given entity (e.g., To the right of the given object is the new subject). This finding is explained in terms of contextual licensing of noncanonical word order. Here, we discuss and tested two alternative accounts of contextual licensing, given-new and partially ordered set relations (Poset). The given-new account claims that noncanonical word order is licensed by the term denoting the given entity preceding the term denoting the new entity. On the Poset account, noncanonical word order is licensed if the preverbal constituent introduces a new entity that stands in a transitive, irreflexive, and asymmetric relation to a given entity. Comprehension times for second premises with spatial adverbs in four different word orders support both accounts of contextual licensing; Poset licensing was stronger than given-new licensing. PMID- 17128617 TI - The World Health Organization World Mental Health Survey Initiative. AB - To present an overview of the World Health Organization World Mental Health (WMH) Survey Initiative. The discussion draws on knowledge gleaned from the authors' participation as principals in WMH. WMH has carried out community epidemiological surveys in more than two dozen countries with more than 200,000 completed interviews. Additional surveys are in progress. Clinical reappraisal studies embedded in WMH surveys have been used to develop imputation rules to adjust prevalence estimates for within- and between-country variation in accuracy. WMH interviews include detailed information about sub-threshold manifestations to address the problem of rigid categorical diagnoses not applying equally to all countries. Investigations are now underway of targeted substantive issues. Despite inevitable limitations imposed by existing diagnostic systems and variable expertise in participating countries, WMH has produced an unprecedented amount of high-quality data on the general population cross-national epidemiology of mental disorders. WMH collaborators are in thoughtful and subtle investigations of cross-national variation in validity of diagnostic assessments and a wide range of important substantive topics. Recognizing that WMH is not definitive, finally, insights from this round of surveys are being used to carry out methodological studies aimed at improving the quality of future investigations. PMID- 17128618 TI - The ESEMeD-WMH project: strenghtening epidemiological research in Europe through the study of variation in prevalence estimates. AB - The European Study of the Epidemiology of Mental Disorders (ESEMeD) project is the first cross-national European survey to collect data on prevalence, risk factors, health-related quality of life and use of services associated with common mental disorders. Representative samples of the non-institutionalized populations of Belgium, France, Germany, Italy, the Netherlands and Spain were interviewed using the CIDI 3.0, totaling 21,425 adult individuals. There was important variation across countries in prevalence estimates of the disorders studied, with Belgium and France in the highest proportions and Italy and Spain the lowest. Comorbidity of mood and anxiety disorders was frequent in all countries. Use of services due to mental health is low in all European countries, but especially low in Italy. Remarkably, only about one in five participants with a mental disorder had consulted the health services in the prior year. We discuss about the possible explanations of cross-national differences in prevalence estimates of mental disorders, and some of the limitations of the data on alcohol abuse and dependence gathered in the study. We conclude that despite the advances in knowledge about determinants and treatment methods, the care of the mentally ill is rather limited in Europe. PMID- 17128619 TI - What is a systematic review? PMID- 17128621 TI - A public health genetic approach for schizophrenia. AB - BACKGROUND: Schizophrenia is a complex disease resulting from the interplay of genetic and environmental factors. However, psychiatric genetics and epidemiology have often worked as independent research fields. AIMS: To review the evidence about the gene-environment interplay involved in the development of schizophrenia. METHODS: Systematic review of medical and psychological databases. RESULTS: On one hand, quantitative and molecular genetics showed high heritability for schizophrenia and identified genes likely to be involved in its pathophysiology. The strength of the association between candidate genes and schizophrenia is however modest, and the need for a more appropriate conceptualization of the genetic risk has been claimed. On the other hand, psychiatric epidemiology described several environmental factors liked with the onset or the course of schizophrenia. The observational nature of epidemiology, however, may hamper inference on causation. Gene-environment correlations and interactions influence the exposure and the vulnerability to the environment, respectively. Current findings suggest that gene-environment correlations and interactions may be common phenomena in the pathophysiology of schizophrenia. The consideration of gene-environment interplay may help to overcome many limitations of genetic and epidemiological studies in psychiatry and suggest innovative preventive and therapeutic strategies. CONCLUSIONS: Taking into account the complexity of schizophrenia pathophysiology, mental health genetics may provide a comprehensive and heuristic model of disease. PMID- 17128620 TI - Cross-cultural studies of personality traits and their relevance to psychiatry. AB - AIMS: This article provides a brief review of recent cross-cultural research on personality traits at both individual and culture levels, highlighting the relevance of recent findings for psychiatry. METHOD: In most cultures around the world, personality traits can be clearly summarized by the five broad dimensions of the Five-Factor Model (FFM), which makes it feasible to compare cultures on personality and psychopathology. RESULTS: Maturational patterns and sex differences in personality traits generally show cultural invariance, which generates the hypothesis that age of onset, clinical evolution, and sex differences in the prevalence of psychiatric disorders might follow similar universal patterns. The average personality profiles from 51 cultures show meaningful geographical distributions and associations with culture-level variables, but are clearly unrelated to national character stereotypes. CONCLUSIONS: Aggregate personality scores can potentially be related to epidemiological data on psychiatric disorders, and dimensional personality models have implications for psychiatric diagnosis and treatment around the world. PMID- 17128622 TI - Mental health system and services in Albania. AB - AIMS: To describe the mental health system in Albania. METHODS: Data were gathered in 2003 and in 2004 using a new WHO instrument, World Health Organization Assessment Instrument for Mental health Systems (WHO-AIMS), designed for collecting essential information on the mental health system of low and middle income countries. It consists of 6 domains, 28 facets and 156 items. RESULTS: The information collected through WHO AIMS covered the key aspects of mental health system in Albania: the mental health policy and the legislative framework, the network of mental health services and the characteristics of the users, the role of the primary health care, the human resources, the public education and the links with other governmental sectors, monitoring and research. CONCLUSIONS: The data collection through WHO AIMS represented a needed step for a better in-depth knowledge of the system and for implementing actions to strengthen the system. Examples of planned actions were the improvement of the mental health component in primary care, a clear shift of resources from mental hospitals to community facilities, an increase of the outpatient care and an expansion of the mental health information system. PMID- 17128623 TI - [The descriptive epidemiology of suicide in Tuscany, 1988-2002]. AB - AIMS: To describe the epidemiology of Suicide in Tuscany according to the triad of time, place and person. METHODS: The 4,764 cases of suicide, defined according to categories E950-E959 of ICD-9 in Tuscany over the period 1988-2002, were obtained from the Tuscan Mortality Register. Mortality indicators were calculated and analyzed. The spatial analysis was carried out by deriving Empirical Bayes Estimates for the 287 municipalities. RESULTS: The crude mortality rate in the 2000-2002 is 7.8 per 100000 population (male: 12.4; female: 3.5). The age standardized rate in the 2000-2002 is 5.8 per 100,000 population (male: 9.6; female: 2.6). The highest risk for suicide, especially in the case of males, are concentrated in the southern hinterland Tuscany, in a cluster of rural municipalities that represent the old mining district of Tuscany. The SMRs according to residential municipality (population per square kilometre), confirm a greater risk of suicide for males residing in rural communities. CONCLUSIONS: The cluster of excessive mortality from suicide in Southern Tuscany could be the consequence of social determinants, related to the urban and social crisis following agriculture decline and mine closure. PMID- 17128624 TI - [Social deprivation and mental health. Replicability and applicability in the Italian context of the resource allocation methods developed in the United Kingdom]. AB - AIMS: Most of the available evidence on the relationship between socioeconomic indicators of social deprivation and patterns of use of mental health services has been produced in the United Kingdom, where the Ministry of Health has developed a resource allocation formula based upon the results of those studies. The main aim of the paper is to evaluate the replicability in the Italian context of such research, and of the resulting allocation strategies. METHODS: Detailed description of the resource allocation method currently adopted in the United Kingdom, whose main purpose consists in reaching the best balance between available funding and patterns of need. Detailed description of resource allocation processes in Italy; discussion of the main methodological and statistical limitations restraining the replicability of the British formula in the Italian context. CONCLUSIONS: There is a growing interest in Italy towards the introduction of evidence-based methods in health decision making, in order to correct the overwhelming influence of political issues. What is needed is a better understanding of the relationship between higher levels of equity in health services access, and their effects in terms of better outcomes. PMID- 17128625 TI - [Family psychoeducational interventions for schizophrenia in routine settings: impact on patients' clinical status and social functioning and on relatives' burden and resources]. AB - OBJECTIVE: This study explored the effectiveness of a psychoeducational family intervention for schizophrenia on patients' clinical status and disability and relatives' burden and perceived support. METHODS: The study has been carried out in 17 mental health centres. In each of them, 2 professionals were trained in a psychoeducational intervention and applied it for six months with families of users with schizophrenia. At baseline and six months later, patients' clinical status and disability, and relatives' burden, social network and professional support were assessed by validated tools. RESULTS: Of the seventy-one recruited families, 48 (68%) completed the intervention. At six months, a significant improvement was found in patients' clinical status and social functioning, as well as in relatives' burden and social and professional support. In particular, the percentage of patients with poor or very poor global social functioning dropped from 50% to 27% at six months. Forty percent of patients and 45% of relatives reported a significant improvement in their social contacts over the intervention period. CONCLUSIONS: The results of this study confirm the hypothesis that psychoeducational family interventions may have a significant effect on social outcome and family burden in schizophrenia when provided in routine conditions. PMID- 17128626 TI - Reliability, validity and acceptability of the WHOQOL-Bref in a sample of Italian psychiatric outpatients. PMID- 17128627 TI - [Effectiveness of hospitalisation in acute psychiatric in-patient units in Lombardy (Italy)]. PMID- 17128628 TI - [On the way to Bethlehem. Visit to the only Palestinian pediatric hospital]. PMID- 17128629 TI - [Annual Meeting of the BeKD e.V. in Mainz. Current topics, lively exchange of ideas and engaging professional politics]. PMID- 17128630 TI - [One day means living 24 hours. The working day at the Sternenbrucke pediatric hospice]. PMID- 17128631 TI - [Long waits for cost approval after clinic discharge]. PMID- 17128632 TI - [A blind mother]. PMID- 17128633 TI - [Instructions for placing a central venous catheter: what is the most effective way to prevent infections?]. PMID- 17128634 TI - [The sick person in art]. PMID- 17128635 TI - [Creating the extrauterine environment for premature infants]. PMID- 17128636 TI - [Optimizing structure and process by introducing patient-centered nursing/primary nursing]. PMID- 17128637 TI - [Position of the DGK e.V. on the letter by Mrs. Gabi Engler and Mrs. Monika Soder, Kinderkrankenschwester no. 9/2006, page 384]. PMID- 17128639 TI - Noninvasive tests for liver fibrosis. AB - There is little doubt that we need fibrosis markers or scoring systems that have proven utility in assessing common hepatic diseases, such as hepatitis C and NASH. Despite multiple research studies, none has yet proved to be of clinical value. The most likely benefit of these tests will be to distinguish between minimal and advanced fibrosis or cirrhosis. Several panels of markers have shown utility in this regard but are not yet established as useful for all hepatic diseases. Until they have been validated and clinically accepted, liver biopsy will continue to be the "gold standard" for assessing liver fibrosis. PMID- 17128640 TI - Management of portal hypertension. AB - Primary prophylaxis is advisable for all patients with moderate-sized or large esophageal varices. After the first episode of bleeding, secondary prophylaxis should be initiated to prevent rebleeding. Multiple treatment modalities are available for each circumstance. Optimal regimens have not yet been established but are under investigation. At present, nonselective beta-blockers are the drugs of choice for primary prophylaxis. In patients with actively bleeding varices, octreotide or, in rare cases, a combination of vasopressin and nitroglycerin is used in conjunction with endoscopic band ligation of the varices. Either a beta blocker, band ligation, or a combination of the two is appropriate for secondary prophylaxis. The combination is often the better choice for patients for whom primary prophylaxis with beta-blockers has failed. As in any treatment situation, the specific approach must be tailored to clinical circumstances. The patient's preferences and willingness or ability to comply with the therapy must be taken into account as well as the physician's expertise. Interventions for hepatic encephalopathy predominantly focus on reducing the amount of ammonia absorbed or endogenously generated in the body. They include correction of precipitating factors, bowel cleansing, and lactulose therapy. In difficult cases, a combination of lactulose and neomycin, metronidazole, or rifaximin is recommended. Because the prognosis for patients with hepatic encephalopathy is generally poor, orthotopic liver transplantation should be considered. PMID- 17128641 TI - Surveillance for hepatocellular carcinoma. PMID- 17128642 TI - Cardiac resynchronization therapy for CHF. The rewards and risks of biventricular pacing. AB - Patients with advanced heart failure continue to experience high morbidity and mortality despite recent progress made with the use of such drugs as beta blockers and angiotensin-aldosterone inhibitors. Cardiac transplantation has severe limitations due to the short supply of organs and the ineligibility of most CHF patients for this therapy. Approved heart-assist devices are cumbersome and costly. Therefore, these devices are currently used mainly in tertiary care centers in a limited number of patients. CRT has been rapidly evolving as a viable and beneficial therapy that is universally applicable by percutaneous method in patients with moderate or severe heart failure. Its relative ease of use and cost-effectiveness make it an attractive option for patients with symptomatic heart failure. Therefore, more physicians are becoming aware of the low threshold for its use. PMID- 17128643 TI - Bladder cancer update. Current evaluation methods and standard of care. AB - Urinary bladder cancer arises in the transitional cell epithelium of the bladder. In the United States, it is the fourth most common tumor in men and the eighth most common tumor in women. Here, Dr Sonpavde reviews the risk factors for bladder cancer, its diagnosis, the latest concepts of its molecular characteristics and metastatic potential, and current strategies to treat this common disease. He focuses in particular on management of transitional cell carcinoma, which is responsible for about 95% of cases. PMID- 17128644 TI - Cutaneous manifestations of diabetes. Signs of poor glycemic control or new-onset disease. AB - Diabetes attacks multiple organ systems, and the skin is not spared its detrimental effects. Pathologic consequences of diabetes, such as vasculopathy, neuropathy, immunologic disturbances, defects in collagen formation, and poor glycemic control, contribute to its numerous cutaneous manifestations. In this article, Drs Tabor and Parlette present the implications of, and treatment options for, infections and dermatoses that appear in established diabetes or that indicate new-onset disease. They also discuss diabetes medications that commonly cause cutaneous reactions. PMID- 17128645 TI - Putting sleep apnea to rest. Tailored therapy reduces fatigue-related risks. AB - Sleep apnea, the periodic cessation of breathing during sleep, is a common and underdiagnosed condition. Treatment can improve quality of life as well as reduce morbidity and mortality from sleep apnea-related hypertension, stroke, and traffic accidents. In this article, the authors discuss types of sleep apnea, diagnostic tools, and treatment strategies for both adults and children. PMID- 17128646 TI - Eating disorders in primary care. A guide to identification and treatment. AB - Awareness of the signs and symptoms of eating disorders is essential for primary care physicians who work with children, adolescents, and young adults. In this article, Drs Kondo and Sokol describe the clinical manifestations of eating disorders such as anorexia nervosa, bulimia nervosa, and binge-eating disorder, which usually occur in adolescence and early adulthood, and pica and rumination disorder, which are more characteristic of infancy and early childhood. The authors also discuss risk factors for and medical complications of eating disorders and present current treatment methods. PMID- 17128647 TI - Detection of diabetic myonecrosis. Complication is often-missed sign of underlying disease. PMID- 17128648 TI - Subacute glaucoma masquerading as migraine. How to avoid the pitfall and make the diagnosis. PMID- 17128649 TI - Better health well in hand. Cell phones have the capacity to more frequently and efficiently connect chronically ill patients with caregivers. PMID- 17128650 TI - Making it work. Are vendors from Mars, clients from Venus? PMID- 17128651 TI - Off-the-chart success. A multi-specialty community healthcare practice implements an EHR system to improve patient care. PMID- 17128652 TI - No clarity in NPI adoption. Confusion abounds as providers try to figure out the nuances of applying for national provider identifiers. PMID- 17128653 TI - The heart of the matter. Whether for the hospital or the office, cardiologists need IT solutions as flexible as they are. PMID- 17128654 TI - How to treat actinic keratosis on hair-bearing skin? PMID- 17128655 TI - Is MRI contraindicated after coronary stent placement? PMID- 17128656 TI - Patient notes: weight loss. PMID- 17128657 TI - First-episode schizophrenia. A window of opportunity for optimizing care and outcomes. AB - The pernicious course of schizophrenia has spurred efforts to identify and effectively treat this condition as early as possible. This assertive therapeutic stance is supported by epidemiologic data suggesting a substantial time lag between onset of illness and therapeutic intervention, and by neurobiologic data suggesting that brain changes present in first-episode psychosis are comparable to those in chronic schizophrenia. The proposal that atypical antipsychotic medications may prevent illness deterioration and/or be a restorative intervention is an appealing, but as yet unproven, hypothesis. Major challenges to maximizing treatment outcomes in first-episode schizophrenia include optimizing timing and effectiveness of pharmacologic interventions, service coordination, and access to care. We present data on the onset and presentation of first-episode schizophrenia and emerging findings about the neurobiology of first episodes, review nonpharmacologic and pharmacologic management, and summarize clinical research data on use of atypical antipsychotics in first episode patients. PMID- 17128658 TI - Health monitoring for patients who have schizophrenia. Summary of the Mount Sinai Conference recommendations. AB - Schizophrenia is associated with several chronic medical illnesses and a reduced life expectancy. This paper summarizes findings and recommendations from "The Mount Sinai Conference," held at the Mount Sinai School of Medicine in New York on October 17-18, 2002, and discusses the implications for improving medical monitoring of patients with schizophrenia who are managed in outpatient settings from the initiation of treatment. The Mount Sinai Conference involved a diverse panel of experts, including specialists on schizophrenia, obesity, diabetes, cardiology, endocrinology, and ophthalmology. Consensus recommendations included baseline measurement and regular monitoring of body mass index, blood glucose, lipid profiles, signs of prolactin elevation or sexual dysfunction, and movement disorders. Information from such measurements should be considered when selecting or switching antipsychotic agents and should trigger an evaluation of medication when abnormalities are detected. PMID- 17128659 TI - Switching in the era of atypical antipsychotics. An updated review. AB - The introduction of the newer atypical antipsychotic medications in the 1990s was the first major change in the psychopharmacologic treatment of schizophrenia in many years. Many patients previously treated with conventional antipsychotics showed marked improvement when switched to one of the newer agents. Yet the principles involved in switching patients to these agents initially posed a challenge for clinicians, with many patients becoming unstable and relapsing during switches. Now, a decade later, switching techniques are better understood, but, with atypical antipsychotics now the standard of care, new challenges have emerged. Surveys confirm a steady decrease in use of older antipsychotics and an increase in use of first-line atypical agents since 1994. Before the newer atypical agents, elective antipsychotic changes in "stable" outpatients were uncommon, but with more choices available, rates of switching, mostly from one atypical agent to another, are now 30%-50% a year in many outpatient clinics, a trend that is likely to continue with the expanding range of target symptoms and growing expectations for better outcomes. Whereas relapse prevention was once considered the primary goal, the focus has moved beyond stability towards recovery. This article focuses on recent developments in switching and highlights differences in the switching profiles of the most recently introduced antipsychotics, ziprasidone and aripiprazole, and compares their switching characteristics with those of the other first-line atypical antipsychotics. PMID- 17128660 TI - Beyond psychopharmacology. Psychosocial strategies for getting the best results when switching antipsychotic medications. AB - Recent effectiveness studies have shown that over a year and a half, 3 of 4 study patients either stopped or changed their antipsychotic medication. This finding highlights the fact that psychopharmacologic treatments do not remain stable over the course of treatment. Our own interpretation is that these findings emphasize that medication decisions cannot be made outside the context of patients' beliefs, life goals, and social network. Especially important is understanding the underlying, often unspoken, hopes and goals that may motivate patients to try a new medication. For best outcomes when changing patient medications, physicians must translate unspoken patient thoughts and feelings into mutually agreed upon definable target symptoms before the switch. This process helps the patient become an active participant in the switching process and encourages patience and perseverance during the lag time before desired improvements may be seen. Concerns about the new medication are best addressed before the switch. Once the decision has been made to switch, the change must await patient psychological readiness to complete a full therapeutic trial of the new medication. The target goals established before the switch can be used to evaluate the success of the medication trial and to compare any improvements with those from the previous medication. It is also important to revisit the initial treatment goals to distinguish primary pharmacologic improvements from secondary psychologic reactions and challenges arising from those improvements. PMID- 17128661 TI - Recovery from a psychiatrist's viewpoint. AB - Recovery is not the same as cure. Recovery from mental illness is the process of having more to life than just illness. It is an ongoing process rather than simply a goal that can be achieved. Recovery from the stigma of mental illness may be as difficult as recovery from the illness itself. Several common, but incorrect, beliefs can interfere with the recovery process. Myths include the belief that the illness has an inherently downhill course, that rehabilitation is useful only after stabilization, and that people with schizophrenia can only work at low-level jobs. People who have schizophrenia have reported that their own process of recovery was helped by their determination to get better, an understanding of the illness, taking personal responsibility, having friends who accept them, an optimistic attitude, and spiritual beliefs that help them find meaning in life. PMID- 17128662 TI - [Epidemiology of osteoporosis in Croatia]. AB - There is no disease registry for osteoporosis and osteoporotic fractures in Croatia. According to the annual report of registered diseases, the number of patients with osteoporosis and osteoporotic fractures increased during the last 10 years. In 2004, 0.70% of all registered diseases in Croatia referred to osteoporosis and osteomalacia. Epidemiological data show that the prevalence of osteoporosis in Croatia is similar to other European countries, while the prevalence and incidence of osteoporotic fractures is somewhat higher. It is estimated that 90 000 men and 77 000 women in Croatia older than 50 years have vertebral fractures. The prevalence is more frequent in older age and in women, although the great number of men (16.2%) were diagnosed as having osteoporosis using ultrasound densitometry. Calcium intake is found to be a dominant risk factor for building-up and maintenance of bone density. PMID- 17128663 TI - [p53, bcl-2 and Ki-67 in the diagnosis of insular thyroid gland cancer. Case report with a review of literature]. AB - The report covers the first case of insular thyroid cancer recorded in Croatian medical literature. A 71-year old female patient presented to our hospital with symptoms of inspiratory stridor. Clinical examination disclosed an expansive tumor mass. US guided FNA indicated anaplastic cancer and total thyroidectomy was accordingly indicated. Pathohistological analysis of H/E biopsies disclosed a tumor mass with a characteristic insular growth pattern of monomorphic tumor cells. Immunohistochemical analysis confirmed diagnosis by a diffuse positive reaction to thyreoglobulin, focally to TTF-1, and scant to cytokeratin, while calcitonin, FVIII, CD34 and LCA were clearly negative. P53 and bcl-2 were expressed in 35.9% and 85% of tumor cells, respectively. The proliferation index for Ki-67 was 4.19. According to our survey we suggest a panel of immunohistochemistry for diagnosing insular cancer. Even if partly present, insular cancer should be mentioned in the pathohistological description because of its prognostic meaning. PMID- 17128664 TI - [Primary pigmented nodular adrenocortical disease as cause of Cushing's syndrome associated with Carney complex]. AB - We report a 11-year-old girl and two 14-year-old boys with Cushing's syndrome due to primary pigmented nodular adrenocortical disease (PPNAD). In these patients, hypercortisolism is a consequence of autonomous cortisol secretion from adrenal glands and is ACTH-independent. Besides PPNAD, the girl had lentigines, spotty pigmentation on her bucal mucosa and lips and she also had schwannoma. One of the reported boys had prolactinoma. Considering this, those two patients fulfill the criteria for Carney complex which is a type of multiple endocrine neoplasia syndromes inherited in an autosomal dominant trait. The other boy had PPNAD but no other obvious signs of Carney complex were noticed. Family study didn't reveal any clinical or laboratory signs of Carney complex in our patients' first relatives. All of our patients underwent bilateral adrenalectomy (in one of the boys laparoscopic surgery was performed). Glucocorticoid and mineralocorticoid substitution has been started. Adrenal glands were macroscopically normal but pathohistological analysis confirmed the diagnosis of PPNAD. PMID- 17128665 TI - [Radiologic imaging of primary bone lymphoma--a retrospective study]. AB - The purpose of our study was to retrospectively review imaging appearances of primary bone lymphoma in 13 patients (six males, seven females; mean age 30.6 years) from 1994 to 2006 on plain radiography, computed tomography (CT), magnetic resonance (MR) and scintigraphy. Lytic-sclerotic pattern was noticed in 26 of 36 bone lesions on plain radiography and in 27 of 33 lesions on CT scanning. On MR, lesions were homogeneous and hypointense on T1 weighted images, and inhomogeneous and hypointense, isointense or hyperintense with respect to fat on T2 weighted images. Increased uptake on bone scintigraphy was noticed in six out of eight patients. The most common location of primary bone lymphoma was the pelvis. Pathologic type was non-Hodgkin's lymphoma in 11 patients and Hodgkin's disease in two patients. Primary bone lymphoma presents as mixed lytic-sclerotic lesions that might cause the soft-tissue mass and bone marrow changes associated with little cortical destruction. PMID- 17128666 TI - [Cardiac natriuretic peptides in heart failure]. AB - Heart failure is a clinical syndrome characterized by abnormalities of cardiac, skeletal muscle, and renal function, stimulation of the sympathetic nervous system, and a complex pattern of neurohormonal changes. Early diagnosis and treatment of heart failure are important factors in reducing morbidity and mortality associated with the disease. During the past several years many studies have demonstrated that natriuretic peptides are the biochemical markers of choice for diagnosing and risk stratification of patients with chronic heart failure. The value of natriuretic peptides has already been recognized by their inclusion in the recent European guidelines for the diagnosis of chronic heart failure. The natriuretic peptides family is comprised of four peptides, each with common 17 amino acid ring structure. The tissue-specific distribution and regulation of each peptide are different. At present, there is no consensus on the most reliable method for natriuretic peptides assay, although non-competitive immunoassay may be more suitable for clinical routine than competitive assay. PMID- 17128667 TI - [Laparoscopic surgery in acute abdominal trauma]. AB - In the past few years, laparoscopy has been increasingly used in patients with acute abdominal trauma. This metod in experienced hands enables direct eye visualisation of abdominal cavity, ensures accurate and early diagnosis and leads to prompt decision on the therapy. Reviewing the literature, laparoscopy in acute abdominal trauma is a safe and accurate procedure to yield diagnosis, enable laparoscopic interventions and help avoid unnecessary laparotomy. In acute abdominal trauma, laparoscopy is mostly used as a diagnostic method, not so often as a therapeutic method, but an expanding range of therapeutic interventions is available. We show our early results with diagnostic and therapeutic laparoscopy from January 1st 2003 to March 31st 2004, stressing therapeutic laparoscopy. We performed 79 laparoscopic explorations in patients with diagnosis of acute abdomen. 7 of them were explored because of acute abdominal trauma. In 3 patients there was no need for intervention after laparoscopic exploration, in 1 patient we electrocoagulates smaller bleeding from the spleen and in other 3 patients we made conversion to open surgery procedure, because of III-IV grade liver or spleen injury. In our group we didn't have injuries of any other internal abdominal organs. Laparoscopic treatment of acute abdominal trauma with spleen or liver injury is applicable in I-III grade liver injury and I-II grade spleen injury, while in greater injuries it is necessary to make conversion to laparotomy. PMID- 17128668 TI - [Cervicogenic proprioceptive vertigo: etiopathogenesis, clinical manifestations, diagnosis and therapy with special emphasis on manual therapy]. AB - Cervical proprioceptive system (CPS) consisting of mechanoreceptors of cervical intervertebral joints, mechanoreceptors of neck muscles and ligament's insertions, muscle spindles located in deep short muscles of cervical spine and sensitive fibers connecting neck's proprioceptors with neurons of cornu posteriori of spinal cord, plays an essential part in maintaining bodily balance. CPS, via tractus spinovestibularis, is connected to vestibular nuclei. Clinical and neurophysiological studies have shown that functional disorders and/or organic lesions of CPS cause identical symptoms as vestibular diseases: vertigo, nystagmus and balance disorders. Dysfunction (functional blockade) of craniocervical joints is the most frequent cause of cervicogenic proprioceptive vertigo (CPV). The constant tension of the capsule of a blocked joint irritates mechanoreceptors protecting the joint's capsules. The increased activity of mechanoreceptors results in confusion of vestibular system. That is, the impulses from the blocked craniocervical joints do not correspond to the impulses from the vestibular organ and other sensory systems that take part in maintaining bodily balance. The disharmony of impulses results in an inadequate vestibulo-spinal and vestibulo-ocular reaction manifesting as vertigo and nystagmus. Hyperactivity of craniocervical mechanoreceptors also causes disturbances in reflex regulation of postural muscle tonus manifesting as "general instability". Knowledge of CPV as a separate clinical entity is important from diagnostical and therapeutical aspect. As it concerns a peripheral vestibular disorder still unknown to a wider circle of physicians, the article describes etiopathogenesis, clinical manifestations, diagnosis and therapy of CPV with special emphasis on manual therapy. PMID- 17128669 TI - [Use of magnetic resonance imaging in the diagnosis and prognosis of multiple sclerosis]. AB - Multiple sclerosis is an autoimmune disease characterized by demyelination and axonal loss. Conventional magnetic resonance imaging allows the demonstration of spatial and temporal dissemination of multiple sclerosis lesions earlier than is possible from clinical assessments. A variety of conventional magnetic resonance imaging protocols, in conjunction with clinical assessment, are now routinely used to increase the accuracy of diagnosis and long-term prognosis of multiple sclerosis. T2-weighted hyperintense lesions are related primarily to increased water content and thus cannot distinguish between inflammation, edema, demyelination, Wallerian degeneration, and axonal loss, whereas the contrast gadolinium-enhanced lesions on T1-weighted images reflect increased blood-brain barrier permeability associated with active inflammatory activity. Conventional magnetic resonance imaging metrics are not sufficiently sensitive to detect invisible brain damage in the normal appearing brain tissue, and they do not show a reliable correlation with clinical measures of disability. However, numerous studies showed that they can improve accuracy in the diagnosis and prognosis of multiple sclerosis. Recently, non-conventional magnetic resonance imaging techniques have been introduced to increase the accuracy of diagnosis and prognosis of multiple sclerosis. Several studies have used brain atrophy, T1 hypointense lesion volume, magnetization transfer imaging, diffusion-weighted imaging and magnetic resonance spectroscopy to test whether the extent and severity of tissue loss in lesions and in normal appearing gray and white matter at the time of clinically isolated syndrome may have diagnostic and prognostic value. These magnetic resonance imaging techniques represent a powerful tool to non-invasively study different pathological substrates of lesions and microscopic tissue changes. Additional short- and long-term prospective studies are requested to establish their value in the diagnosis and prognosis of multiple sclerosis. PMID- 17128670 TI - [Neurofibromatosis type 2 (central neurofibromatosis or bilateral acoustic neuromas, vestibular schwannomas): from phenotype to gene]. AB - Neurofibromatosis type 2 (NF2) is an autosomal dominant disease that predisposes to bilateral vestibular schwannomas (neurinomas), other central and peripheral nervous system tumours (multiple meningeomas and neurofibromas) and ocular abnormalities (cataract). The NF2 tumour suppresor gene is localised on chromosome 22q12 and encodes protein called schwannomin or merlin which is related to a family of cytoskeleton-to-membrane proteins linkers ERM (ezrin radixin-moesin proteins). About 50% of all cases are new germline mutations, although about 20% of apparently sporadic cases represent somatic mosaicism. The majority of observed germline NF2 mutations are point mutations which result in schwannomin with an altered or absent C-terminal domain. NF2 has a variable clinical presentation, with two basic types: severe type having early onset and progressive growth of tumors and the milder type having later onset and less aggressive course. The genotype-phenotype correlations indicate a greater variability of clinical disease expression. In this paper we discuss the epidemiology, genetic and clinical characteristics, diagnostic criteria, investigations, screening for risk persons and recommendations for care and therapy of patients with NF2. PMID- 17128671 TI - [Anaphylactic reaction as a side-effect of administration of general anesthetics and neuromuscular blocking agents]. AB - The incidence of anaphylactic reactions during anesthesia is between 1:5000 and 1:25000 anesthetics. During the IgE-mediated anaphylactic reaction mast cells release proteases such as tryptase, histamine and vasoactive mediators. The release of mediators from the mast cells and basophils is responsible for the immediate clinical manifestations of anaphylaxis. Anaphylactoid reactions can be caused directly by a release of histamine and other mediators from mast cells and basophils and they don't depend on interaction of IgE antibodies with antigen. The most frequent agents that cause anaphylactic and anaphylactoid reactions during anesthesia are neuromuscular blocking agents (among them the highest percent refers to rocuronium and succinylcholine), some general anesthetics, antibiotics, blood and blood products, opioids and latex. Increased tryptase concentration in serum is a marker for systemic mast cell activation. Skin tests (in vivo) are used for verification of specific hypersensitivity to drugs in patients after anaphylactic reaction. In vitro tests prove the presence of specific IgE antibodies for drugs. The plan for the treatment of anaphylactic reactions must be established before the event. Airway maintenance, 100% oxygen administration, intravascular volume expansion and epinephrine are essential to treat the hypotension and hypoxia that result from vasodilatation, increased capillary permeability and bronchospasm. As soon as the diagnosis has been made the adrenalin should be given intravenously 1 to 3 ml of 1:10000 aqueous solution (0.1 mg/ml) over 10 minutes. Prevention is possible with methylprednisolone 125 mg i.v. 1 hour before administering of anesthetics and neuromuscular blocking agents with or without antihistaminic chlorpiramine-chloride 1 amp i.v. few minutes before anesthesia. PMID- 17128672 TI - [Characteristics of a new bladder cancer classification]. PMID- 17128673 TI - Aging and mental illness. What we don't know. PMID- 17128674 TI - Pharmacotherapy strategies for treatment-resistant depression. PMID- 17128675 TI - Developing programs for older adults in a faith community. AB - I have worked in all areas of nursing, from staff and head nurse positions to faculty and administrative positions, but my work as a parish nurse, particularly with our community of older adults, is one of the most challenging and rewarding areas of nursing practice I have ever experienced. Recently, one of our independent older adult parishioners, Marge, suddenly had a marked decrease in her ability to function, which was precipitated by two trips to the hospital and surgery. She went from living independently to living in a skilled care unit, no longer able to walk or perform activities of daily living without assistance. I had known Marge for only 2 years. When she became ill, I visited her weekly during the last 4 months of her life, as she had no family in the area. During my visit one evening as I helped Marge get ready for bed, she left me with a very poignant memory. After sharing some hair-raising stories about her care at this facility, she said, "Georgine, you're my best friend. You're the only one I can tell these things to and I know you understand." PMID- 17128676 TI - Interview with Mary Starke Harper, PhD, RN, FAAN. PMID- 17128677 TI - Psychotherapy with older adults: benefits and barriers. AB - Psychotherapeutic treatments are known to be effective in older adults with mental illnesses. Depression, anxiety, and complicated bereavement must be considered psychiatric illnesses, rather than normal consequences of aging. To improve the rate of psychotherapy use among older adults, stigma surrounding psychiatric treatment, as well as misconceptions that older adults are incapable of change, must be unlearned and eliminated. PMID- 17128678 TI - Psychosocial interventions for disruptive symptoms of dementia. AB - An estimated 6% to 8% of the adult population age 65 and older and more than 30% of those age 85 and older are affected by a dementing disorder. The annual direct and indirect cost of caring for the 4.5 million people with Alzheimer's disease in the United States is estimated to be at least $100 billion. By 2030, when the entire Baby Boomer generation is age 65 and older, the increased number of people with Alzheimer's disease could exceed the ability to absorb the added cost. Both professional and familial caregivers should be familiar with the treatment interventions that are most effective in reducing the disruptive behavioral and psychological symptoms of dementia. This article discusses the various kinds of dementia, their associated symptoms, and the psychosocial treatment options that have been found to be effective in alleviating the effects of the disease on both people with dementia and their caregivers. Psychosocial strategies can be divided into four major subgroups: communication techniques, behavioral strategies, environmental modifications, and caregiver education. The ultimate goal is to optimize functioning of people with dementia and minimize caregiver strain. PMID- 17128679 TI - Elder abuse and neglect. AB - Elder abuse and neglect is a critical health care issue that must be brought to the attention of health care providers and older adults' family members. Adults older than 65 who live at home or in long-term care facilities may be at risk for abuse. Nurses should be aware of the causes, screening questions, symptoms of abuse, and resources in the community. Armed with information and a better understanding about the issue, nurses can minimize the devastating effects of abuse on older adults and their families. PMID- 17128680 TI - Disparities in race/ethnicity and gender in in-hospital mortality rates for coronary artery bypass surgery patients. AB - BACKGROUND: While dramatic progress has been made lowering in-hospital mortality for coronary artery bypass graft surgery (CABG), few comprehensive studies have been done that include Caucasian, African-American, Hispanic and Asian American/Pacific-Islander CABG inpatients and simultaneously evaluate the influence of gender. This study, analyzing five years of national data for 1.2 million CABG admissions, examines trends in in-hospital CABG mortality rates for gender and four racial/ethnic categories for CABG patients. METHODS: Using data from the Health Care Utilization Project (HCUP) for 1998-2002, 1.2 million CABG admissions were analyzed using descriptive and logistic regression analyses to evaluate the extent of the disparities in in-hospital CABG mortality rates. HCUP is a sample of nearly 1,000 hospitals from 35 states designed by the Agency for Healthcare Research and Quality (AHRQ) to approximate a 20% stratified sample of the nation's community hospitals: approximately 94% of all hospital discharges in the United States. RESULTS: Although significant progress has been made in recent years in lowering in-hospital CABG mortality, after controlling for relevant patient and socioeconomic factors, female CABG patients, regardless of their racial/ethnic group, still experience significantly higher in-hospital mortality rates than their male counterparts. Additionally, among these racial and ethnic groups, black CABG patients, whether male or female, continue to experience significantly worse in-hospital mortality rates than other races/ethnicities. CONCLUSIONS: The declines in CABG in-hospital mortality rates have not been equal across race/ethnicity and gender. PMID- 17128681 TI - Multiple lipid scoring system for prediction of coronary heart disease risk: application to African Americans. AB - BACKGROUND: Clinicians often obtain a panel of lipids but then only use low density-lipoprotein (LDL) cholesterol to make clinical decisions. We previously described the multiple lipid measure, a strategy that integrates information about seven lipid measures. Our current inquiry uses the multiple lipid measure to create a scoring system and validates that system in a second cohort. METHODS AND RESULTS: A scoring system that uses total cholesterol, high-density lipoprotein (HDL) cholesterol, LDL cholesterol and triglycerides was developed and tested. African-American participants of the Atherosclerosis Risk in Communities (ARIC) Study were used to validate the multiple lipid measure score. For nonsmokers, scores > or = 2 had a hazard ratio of 4.25 (95% CI 1.92-9.40) compared to reference scores of < or = -3 in adjusted survival analysis predicting incident coronary heart disease risk in the ARIC. The best conventional single lipid measure for nonsmokers was LDL cholesterol. Compared to LDL cholesterol <100 mg/dl, those with LDL cholesterol > or = 160 mg/dl had a hazard ratio of 2.31 (95% CI 1.13-4.75). For current smokers, the best conventional lipid measure was the total cholesterol/HDL cholesterol ratio, which was similar in predictive ability to the multiple lipid measure score. However, the multiple lipid measure score predicted an additional 10% of the cohort at risk compared to the total cholesterol/HDL cholesterol ratio. CONCLUSIONS: The use of the multiple lipid scoring system improves the assessment of incident coronary heart disease risk and may have utility for clinicians in integrating lipid values. PMID- 17128682 TI - Plasma osteopontin levels are elevated in non-ST-segment elevation acute coronary syndromes. AB - BACKGROUND: The regions of ruptured atherosclerotic plaques have numerous macrophages. Osteopontin that modulates macrophage function has been shown in atherosclerotic plaques. We aimed to study the plasma levels of osteopontin in patients with unstable angina or non-ST-seg ment elevation myocardial infarction (NSTEMI) and the rela tionship between osteopontin and the extent of the coronary artery disease (CAD). METHODS: We studied 65 patients with unstable angina or NSTEMI, 25 patients with stable angina and 18 patients as the control group. The extent of coronary artery stenosis was determined by the number of vessels with >50% stenosis. Plasma osteopontin concentrations were measured from the blood samples that were drawn immediately after admission to the emergency department in unstable angina/NSTEMI patients and before the coronary angiograph in the stable angina and control groups. RESULTS: The plasma osteopontin concentration was (495 118 ng/ml) significantly higher in the patients with unstable angina/NSTEMI compared to the stable angina group (319 106 ng/ml) and control group (125+/-54 ng/ml) (p=0.0001 The plasma osteopontin levels were lower in the patients with stable angina pectoris who had one-vessel disease compared to those with two-vessel disease (p=0.01). How ever, in the unstable angina/NSTEMI group, the plasma osteopontin levels were statistically not different among the patients with one-vessel, and two-vessel and three-vessel disease (p=NS). There was no correlation between the plasma osteopontin levels and the extent of coronary stenosis. CONCLUSIONS: The plasma osteopontin levels are elevatedin patients with unstable angina/NSTEMI, but there appears to be no correlation with the extent of CAD. These results ma suggest that osteopontin may have a role in the pathobiology of ACS. PMID- 17128683 TI - Management and in-hospital outcome of patients with first episode of acute myocardial infarction: impact of diabetes mellitus. AB - The aim of this study is to compare clinical characteristics, management, complications, in-hospital outcome and mortality of diabetic and nondiabetic patients after the first episode of acute myocardial infarction (MI). This retrospective case-matched study included 60 diabetic and 55 non-diabetic patients. Medical information about patients was retrieved from the hospital recordings. Diabetic patients were older, were more hypertensive and had more coronary artery disease history than nondiabetic patients. The frequency of use of acute reperfusion therapy modalities was not statistically different between the two groups. Diabetes was a significant determinant against the use of beta blockers (OR=0.26; 95% CI 0.06-0.95) but in favor of angiotensin converting enzyme inhibitors (OR=3.3; 95% CI 1.17-9.36), whereas diabetes did not influence the use of other drugs. Diabetic patients had more complications than nondiabetic patients (40.0% and 16.3%, respectively, p=0.005). In-hospital mortality rate for diabetic patients was comparable to nondiabetic patients (16.7% and 10.9%, respectively, p=0.373). Pharmaceutical treatment regimens, particularly beta blockers, are underutilized after acute MI in diabetic patients. More frequent use of these regimens will improve the high complication and mortality rates in diabetic patients as well as in nondiabetic patients. PMID- 17128684 TI - The pattern and prognostic features of QT intervals and dispersion in patients with acute ischemic stroke. AB - BACKGROUND: Ischemic stroke, which is perhaps the commonest subtype of stroke, is associated with electrocardiographic (ECG) changes. Some of these changes have been thought to be due either to the stroke state itself or pre-existing heart disease. Some, particularly QT intervals, have been associated with increased mortality. OBJECTIVE: The aim is to investigate the pattern of QTcmax, QTd and QTcd in patients with ischemic stroke and to compare these changes in patients without pre-existing heart disease in order to determine their prognostic importance. METHODS: Sixty-four patients with acute ischemic stroke were compared with 60 controls observing the various ECG changes. Patients without pre-existing heart disease were isolated and compared with the total cohort. RESULTS: Thirty five (54.7%) of the patients had ischemic-like ECG changes made up of ST depression (29.7%), T-wave inversion (21.8%) and U wave (9.3%). Twenty-eight (43.8%) had QTcmax prolongation. Twenty-four (37.5%) of the patients had no pre existing heart disease. The QT was similar when compared with the total cohort except in QTcmax, where there was significant difference (447.3+/-72.2 vs. 408.6+/-40.3 msecs). Mortality rate of the total cohort at 28.1% was significantly higher than in those without pre-existing heart disease at 8.3%, suggesting that presence of pre-existing heart disease contributed to mortality. QTcmax (r=0.293 p=0.045) and days on admission (r=-0.543 p=0.001) were the other variables that correlated with mortality in the total cohorts. CONCLUSION: Ischemic-like and repolarisation ECG changes are common in our patients with acute ischemic stroke. These changes tend to be due to pre-existing heart disease rather than the stroke state. PMID- 17128685 TI - Clinically diagnosed diabetic neuropathy: frequency, types and severity. AB - OBJECTIVE: Studies of frequency of occurrence of diabetic neuropathy are few, and available studies were limited to the southern part of Nigeria. The objectives of the study were to determine the frequency of occurrence and grades of diabetes peripheral neuropathy using clinical measures. PATIENTS AND METHODS: Consecutive patients with diabetes mellitus attending the Jos University Teaching Hospital were recruited as the study population, including 120 diabetics and 60 age matched, nondiabetic controls. A standard proforma based on the Michigan Neuropathy Screening Instrument (MNSI) was employed to screen for diabetic neuropathy. RESULTS: The frequency of occurrence of diabetic peripheral neuropathy was 75%. For the specific types of peripheral neuropathy, sensorimotor neuropathy was the commonest (40.4%, chi(2)=29.1; p<0.001). There was no significant difference, with severity of peripheral neuropathy among diabetics, when compared by gender. (Chi square=3.03, P value=0.081). CONCLUSION: The frequency of occurrence of peripheral neuropathy among diabetics in Jos University Teaching Hospital from this study is rather high. PMID- 17128686 TI - Cultural competency training for third-year clerkship students: effects of an interactive workshop on student attitudes. AB - With an increasing awareness of health disparities, medical schools are challenged to develop training in cultural competency for their students. We developed and evaluated the effectiveness of an interactive workshop designed to improve third-year students' attitudes, beliefs and cross-cultural communication skills. METHODS: At the start of a six-week required family medicine clerkship, 196 medical students participated in small group (20-24 students) workshops. Didactics included facts about health disparities and a model of cultural competency. During a skill-building component, students were exposed to live vignettes portraying ineffective and effective cross-cultural doctor-patient interactions. Impact on students' attitudes, perceived bias and knowledge of techniques was assessed by comparing pre- and postworkshop scores. RESULTS: Participants increased their cultural awareness on most items of a cultural awareness scale. Fifty-five-, 71- and 66% of the sample agreed or strongly agreed the program was valuable, appropriate and effective, respectively. Conversely, only 17-, 6- and 9% of the sample disagreed or strongly disagreed, respectively. CONCLUSIONS: A workshop for third-year students led to an increase in cultural awareness and was considered appropriate and valuable. Further study, including longitudinal training and evaluation, is needed regarding effective methods to increase cultural competence in clinical practice. PMID- 17128687 TI - Incidence of pulmonary hypertension in patients with chronic myeloproliferative disorders. AB - STUDY OBJECTIVE: To assess the incidence of pulmonary hypertension (PH) in patients with chronic myeloproliferative disorders (CMPD). METHOD: Twenty-seven patients with a diagnosis of CMPD were included in the study. Patients were excluded if they had a secondary cause of PH. Diagnosis of PH was established if right ventricular systolic pressure (RVSP) by transthoracic echocardiography (TTE) was >35 mmHg. RESULTS: Diagnosis of PH was established in 14 out of 27 patients. Two patients were excluded from analysis because of poor ejection fraction on TTE, resulting in a final diagnosis of PH in 12 of 25 (48%) patients. Of these 25 patients, seven of nine with essential thrombocytosis (ET), five of 14 with polycythemia vera (PV), and 0 out of two with chronic myeloid leukemia (CML) had PH. All patients were asymptomatic at the time of their most recent visit. There was no relationship between PH and age at diagnosis, duration of disease, platelet count and hematocrit at diagnosis or during follow-up, both for the entire cohort or for specific diagnosis of ET or PV. CONCLUSION: Pulmonary hypertension appears to be common in patients with CMPD. Further studies are needed to evaluate the impact of treatment on PH and long-term survival in these patients. PMID- 17128688 TI - Public health preparedness of health providers: meeting the needs of diverse, rural communities. AB - Meeting the needs of public health emergency and response presents a unique challenge for health practitioners with primary responsibilities for rural communities that are often very diverse. The present study assessed the language capabilities, confidence and training needs of Texas rural physicians in responding to public health emergencies. In the first half of year 2004, a cross sectional, semistructured survey questionnaire was administered in northern, rural Texas. The study population consisted of 841 practicing or retired physicians in the targeted area. One-hundred-sixty-six physicians (30%) responded to the survey. The responses were geographically referenced in maps. Respondents reported seeing patients with diverse cultural backgrounds. They communicated in 16 different languages other than English in clinical practice or at home, with 40% speaking Spanish at work. Most were not confident in the diagnosis or treatment of public health emergency cases. Geographic information systems were found useful in identifying those jurisdictions with expressed training and cultural needs. Additional efforts should be extended to involve African American/Hispanic physicians in preparedness plans for providing culturally and linguistically appropriate care in emergencies. PMID- 17128689 TI - Mobile health unit for minority obesity education: local residents' attitudes and perceptions. AB - OBJECTIVE: To provide educational material to the Bronx community in their places of recreation as well as uncover attitudes and perceptions about the obesity epidemic. METHODS: Our medical team equipped a mobile health unit, called Vehicle Assisted Nutrition (VAN), with educational material and drove it to seven recreation sites in the Bronx. At these sites, participants completed a short survey, and our staff took body mass index (BMI) measurements. The medical team also distributed patient education materials pertaining to lifestyle modification and obesity. BMI and survey responses were compared and analyzed with statistical software. RESULTS: Obesity rates at these sites averaged 29.9% normal weight, 40.2% overweight, 18% class-1 obese and 11% class-2 obese or greater. Only 22.2% of normal weight, 23.3% of overweight and 43.6% of class-1 obese respondents stated that their weight was a problem (p<0.001). Most (80.6%) of the class-2 obese or greater respondents, however, stated that their weight was a problem (p<0.001). More than 44% of normal weight, 59.4% of overweight, 71.3 of class-1 obese and 90.3% of the class-2 obese or greater participants stated they were willing to do something about it (p=0.006). Of those surveyed, 37.5% of the normal weight, 54.7% of the overweight, 56.9% of the class-1 obese and 63.6% of the class-2 obese or greater indicated that they would like our help (p=0.035). More than 66% of normal weight, 41.8% of the overweight, 56.9% of the class-1 obese, and 68.9% of the class-2 obese or greater stated that they would be interested in a class to lose weight (p=0.008). CONCLUSION: Medical practitioners need to recognize the role of patients' attitudes about weight loss and clarify for patients the definitions of exercise. PMID- 17128690 TI - A case-control study of factors associated with HIV infection among black women. AB - OBJECTIVE: To identify social, behavioral and epidemiologic factors associated with HIV infection among HIV-infected and HIV-uninfected black women residing in North Carolina. DESIGN: A case-control study conducted in August 2004 in North Carolina. METHODS: Cases were 18-40-year-old women with HIV infections diagnosed from 2003-2004. Controls were 18-40-yearold, HIV-negative heterosexually active women recruited from HIV testing sites. Five focus group discussions were also conducted with women not participating in the case-control study. RESULTS: Multivariate analyses of 31 cases and 101 controls showed that HIV-positive women were more likely to receive public assistance [adjusted odds ratio (aOR) 7.3; 95% confidence interval (CI) 2.1, 26.0], to report a history of genital herpes infection (aOR 10.6; 95% CI 2.4, 47.2), and were less likely to have discussed a variety of sexual and behavioral issues relevant to risk of HIV infection with their male partners (aOR 0.6; 95% CI 0.4, 0.8). Focus group participants indicated that financial and social demands created competing challenges for making HIV prevention a priority. CONCLUSIONS: Inadequate communication between black women and their sexual partners may create barriers to sexual and behavioral risk reduction. A multidimensional approach that addresses both biological factors such as herpes infection and socioeconomic factors may be needed to reduce HIV transmission in this population. PMID- 17128691 TI - Low prevalence of hepatitis-C viral seropositivity among patients with type-2 diabetes mellitus in a tertiary hospital. AB - Recently, there have been increasing reports of high prevalence of hepatitis-C virus (HCV) in patients with type-2 diabetes, mostly in western nations. This suggests that type-2 diabetic patients could be considered to be at special risk of acquiring HCV and possibly that diabetes has an etiological relationship with HCV. Ninety patients with type-2 diabetes attending the medical outpatient clinic of the University College Hospital (UCH) and 90 nondiabetic controls with comparable age, sex and risk factors of exposure to HCV were recruited into the study. All subjects were screened for anti-HCV using a third-generation rapid enzyme immunoassay (Dialab anti-HCV cassette). Data were analyzed using Student's t test, Chi-squared test and Fisher's exact test. None of the diabetic patients tested positive for anti-HCV, while 1.1% of the control group tested positive for anti-HCV. There appears to be low prevalence of anti-HCV among type-2 diabetic patients in UCH Ibadan, and therefore no demonstrable risk of HCV in our patients. PMID- 17128692 TI - Body mass index as a prognostic factor in endometrioid adenocarcinoma of the endometrium. AB - OBJECTIVE: To determine if body mass index (BMI) influences tumor expression of HER-2/neu, estrogen and progesterone receptors (ER/PR), and survival in women with endometrial adenocarcinoma. METHODS: Patients diagnosed between January 1992 and December 2001 with endometrioid adenocarcinoma of the uterus were identified. Clinical and pathologic data were retrospectively collected. HER-2/neu, estrogen and progesterone receptor expression were determined by immunohistochemistry. Differences in these variables and other prognostic factors were analyzed and correlated with effect on survival. RESULTS: One-hundred-sixty-five patients were included in this analysis. Lower BMI was associated with high stage (p=0.04) and HER-2/neu expression (p=0.04). Black race, high grade, high stage and lack of ER/PR expression were all associated with decreased survival. Despite having better prognostic factors, women with a BMI >25 had a lower survival than women with a BMI <25 (p=0.36). When five-year survival rates were calculated for BMI category and stratified by prognostic factors, for almost every high risk factor, survival was lower in overweight patients. CONCLUSION: In patients with endometrioid adenocarcinoma, low BMI is associated with high stage and tumor expression of HER-2/neu. Despite better prognostic factors, overweight women experience poorer survival. PMID- 17128693 TI - The effect of physician-patient discussions on the likelihood of prostate specific antigen testing. AB - Many medical and professional organizations agree that men should discuss the advantages and disadvantages of testing for prostate-specific antigen (PSA) with their physicians before undergoing testing. In the 2000 National Health Interview Survey, men who had undergone a PSA test in the past were asked about their use of this test and discussions they had with physicians regarding its advantages and disadvantages. Among a group of 2,188 black and white men aged 40-79 years with no history of prostate cancer and a history of testing for PSA, we examined whether physician-patient discussions mediated the relationship between race and PSA testing. We specified that the test had to be their most recent one and part of a routine physical examination or screening test. We compared those tested within the past two years with those tested >2 years. Almost two-thirds of the men previously had discussions with their physicians about the advantages and disadvantages of the PSA test. Older men, college graduates, those living in the midwest and those with health insurance were more likely to have been tested recently. Discussion with a physician was found to mediate the relationship between race and PSA testing during the past two years. Black men were initially found to be more likely than white men to have been screened recently [odds ratio (OR)=1.45; 95% confidence interval (CI) 1.01-2.07], but in the full model race was no longer significant (OR=1.41; 95% Cl 0.98-2.03). Discussions about PSA testing were associated with more recent PSA screening (OR=1.38, 95% CI 1.05 1.82). These findings suggest that: 1) the relationships among race, physician discussions and PSA testing may need to be examined in more complex ways, and 2) the physician has an important role in men's decision to consider PSA testing. PMID- 17128694 TI - Postexercise insulin action in African-American women. AB - African Americans are more insulin resistant than Caucasians. A single bout of moderate-intensity exercise reduces insulin resistance in sedentary Caucasian individuals. The impact of a single bout of exercise on insulin resistance has never been studied in African Americans. The purpose of this experiment was to evaluate the impact of a single bout of exercise on insulin resistance in African American women. DESIGN: Insulin resistance was assessed in 10 sedentary, over weight or obese African-American women during a sedentary and exercise condition over a two-day period. During the sedentary condition, participants fasted overnight and sat quietly in the laboratory for 75 minutes. During the exercise condition, participants completed 75 minutes of brisk walking on a treadmill. Ninety minutes following each condition, participants completed an oral glucose tolerance test (OGTT). Three-and-a-half hours later, subjects consumed a standardized meal [meal tolerance test (MTT)]. RESULTS: The insulin response to the OGTT was 18% lower (p=0.046), and insulin sensitivity was 18% higher (p=0.042) in the exercise condition compared to the sedentary condition. There were no differences between conditions following the MTT. CONCLUSIONS: These results indicate that overweight/obese, sedentary, insulin resistant African American women had a significant improvement in insulin sensitivity from 75 minutes of brisk walking. PMID- 17128695 TI - Cognitive decline in older Mexican Americans with diabetes. AB - OBJECTIVE: To examine social, demographic and health factors associated with cognitive decline over a seven-year period among older Mexican Americans with diabetes. METHODS: A population-based sample of 808 noninstitutionalized Mexican Americans aged >65 years with diabetes who had a Mini-Mental State Examination (MMSE) >17 at baseline from the Hispanic Established Population for the Epidemiological Study of the Elderly (H-EPESE). Measurements included sociodemographics, diabetic treatment received (oral hypoglycemic or insulin), self-reported medical conditions, self-reported diabetes-related complications, high depressive symptoms and ADL limitations. RESULTS: The mean MMSE score at baseline was 25.3 + (SD=3.7). The rate of decline in cognitive function (MMSE) during the follow-up period was 0.37 point per year. Using general linear mixed models, we found that being male, and having high depressive symptoms and diabetic complications (kidney impairment, circulation problems or limb amputation) were factors significantly associated with greater declines in MMSE score over time. CONCLUSION: Circulation problems, kidney impairment and depression are the major factors associated with cognitive decline in older Mexican Americans with diabetes. PMID- 17128696 TI - Acute eosinophilic pneumonia accompanied by mediastinal lymphadenopathy and thrombocytopenia. AB - Acute eosinophilic pneumonia, which was described in 1989, is thought to represent a hypersensitivity reaction to unidentified inhaled antigens. Here, we present a case of a marble mine worker with acute eosinophilic pneumonia complicated with mediastinal lymphadenopathy, neutrophilia, thrombocytopenia and acute respiratory distress syndrome. PMID- 17128697 TI - A case of bilateral psoas abscesses and lumbar osteomyelitis due to recurrent salmonella infection. AB - Psoas abscess and lumbar osteomyelitis due to salmonella infection is very rare, although it is frequently seen all over the world. These two complications have severe clinical progress, poor prognosis and high mortality. Here, we report a case of salmonellosis presenting with bilateral multiple psoas abscesses and lumbar osteomyelitis, which resolved completely following medical treatment and percutoneous drainage of abscess. PMID- 17128698 TI - Successful conservative management of Boerhaave's syndrome with late presentation. AB - In this report, we describe the diagnostic, clinical and therapeutic aspects of a 22-year-old man who was diagnosed with a case of Boerhaave's syndrome (BS) 86 hours after acute onset of chest pain and dyspnea following forceful vomiting due to excessive ingestion of alcohol. Early diagnosis and prompt treatment is critical in BS due to the high mortality rates related to delay in diagnosis. We think the main criterion of prognosis is not only the free interval between diagnosis and treatment but the clinical form. We suggest conservative management, including intrathoracic lavage and close monitoring in patients with late presentation with a distal esophageal tear. PMID- 17128699 TI - Higher prevalence of monoclonal gammopathy of undetermined significance in African Americans than whites--the unknown role of underlying HIV infection. AB - The age-adjusted prevalence rate of monoclonal gammopathy of undetermined significance (MGUS) is three-fold higher in African Americans than whites. Similarly, there is a higher preponderance of multiple myeloma (MM) in African American patients. Since the risk of progression of MGUS to MM is equal in both races, identification of exogenous and genetic risk factors of MGUS [such as genetic pre-disposition; diet; and chronic antigenic exposure related to sexually transmitted diseases, including human immunodeficiency virus (HIV) infection] is essential for unraveling the etiology of the racial disparity for MM. HIV infection, a well-documented risk factor for MGUS, is more frequent in African American patients. Future epidemiologic studies dealing with plasma cell disorders should carefully examine the relationship between race, HIV infection status, prevalence of MGUS and its ultimate progression to MM. PMID- 17128700 TI - The Massachusetts health insurance law: providing health insurance to all. AB - There have been increasing rates of uninsured patients in the United States over the past few decades. Despite this growing problem, little progress has been made to decrease the rate of growth of uninsured patients or to provide affordable insurance coverage to those who are unable to maintain insurance coverage throughout the year. The legislature in the Commonwealth of Massachusetts has enacted legislation that requires health insurance for all in the same manner that automobile insurance is required in many states. This bold comprehensive legislation augments current Medicaid and state child health insurance program coverage and expands coverage options for part-time workers, those employed by small companies or those employed at lower-income jobs. PMID- 17128701 TI - Demographic variations and clinical associations of episiotomy and severe perineal lacerations in vaginal delivery. AB - Primiparity, birthweight, operative delivery and obstetrical complications contribute to episiotomy and severe perineal lacerations. Episiotomy correlates with Hispanics, while African Americans correlate with severe perineal lacerations. OBJECTIVE: The purpose of this study was to identify risk factors for both episiotomy and severe perineal lacerations in a large population from a single institution. STUDY DESIGN: This was a review of 66,224 vaginal deliveries of African Americans or Hispanics delivering between 25-44 gestational weeks between 1981-2001. Univariate and multiple regression analysis were done as indicated. RESULTS: Independent predictors of episiotomy were: primiparity eight fold, forceps delivery seven-fold, vacuum delivery five-fold, shoulder dystocia 3.6-fold, macrosomia 1.8-fold, epidural analgesia 1.6-fold, postdates 1.5-fold, Hispanics 1.4-fold. Independent predictors of severe perineal lacerations were; macrosomia seven-fold, episiotomy 4.5-fold, primiparity 4.4-fold, shoulder dystocia 3.6-fold, average birthweight 3.5-fold, forceps delivery 2.6-fold, vacuum delivery two-fold, epidural analgesia two-fold, African-American 1.5-fold. Nonreassuring fetal heart rate patterns, meconium and cord accidents appeared protective. CONCLUSION: Primiparous women with larger babies undergoing operative delivery with epidural analgesia are at risk for both episiotomy incisions and severe perineal lacerations. Though Hispanics are more likely to have an episiotomy, they are at significantly less risk for severe perineal lacerations compared to African Americans. Even though episiotomy is independently associated with severe perineal laceration, other factors such as macrosomia and primiparity are as important. PMID- 17128702 TI - "I have bad news ... you've got cancer". PMID- 17128703 TI - Bisphosphonate-associated osteonecrosis of the jaw: questioning the proposed protocols. PMID- 17128704 TI - The realities of periodontitis: dentistry's role. PMID- 17128705 TI - Enamel remineralization: the medical model of practicing dentistry. PMID- 17128706 TI - Shade selection and communication: four pieces to a puzzle. PMID- 17128707 TI - Single crown tooth preparation: the 80/20 rule. PMID- 17128708 TI - Retreatment of a failed endodontic case. PMID- 17128709 TI - Stable implant overdentures. PMID- 17128710 TI - Simplifying divergence challenges of a combination root and implant overdenture attachment case. PMID- 17128711 TI - A splint-supported porcelain pontic after adult orthodontic treatment. PMID- 17128712 TI - The role of maxillary development in dental-facial aesthetics: case reports. PMID- 17128713 TI - Taking the guesswork out of staff compensation: a scientific and human approach. PMID- 17128714 TI - Uncommon sense. PMID- 17128715 TI - Work smarter! Let your computers work harder! Five free computer habits to increase practice profitability. PMID- 17128716 TI - Staged periodontal plastic surgery in the management of patients with complex dental problems. PMID- 17128717 TI - Three-dimensional cone beam computerized tomography in dentistry. PMID- 17128718 TI - Buyers' guide to all-ceramic systems. Restoring the appearance of the human tooth. PMID- 17128720 TI - Buyers' guide to LED curing lights. LED curing lights are basic tools of the profession. PMID- 17128719 TI - Buyers' guide to air-driven handpieces. Handpieces play an important role in practices. PMID- 17128721 TI - Should you sign a "consent to settle"? PMID- 17128723 TI - Competition--just a phone call away. PMID- 17128722 TI - Can expense sharing work? PMID- 17128724 TI - The day Joy died. PMID- 17128725 TI - Practice 911. What you can learn from three practice management emergencies. PMID- 17128726 TI - Ready, set, swipe. PMID- 17128727 TI - What would you do? Splitting pills. PMID- 17128728 TI - Exclusive earnings survey. How are you doing? PMID- 17128729 TI - Assessing hospitals' responsibility to provide unremunerated services. AB - Hospitals typically absorb the costs of unremunerated health care services; however, budget cuts make this no longer feasible. This article identifies important ethical and legal dimensions of a hospital's duty regarding unremunerated services in a climate of scarce resources. While, except in certain emergency situations, a hospital has no obligation to provide services to those who cannot pay, some guidelines are offered to help hospitals respond to requests for unremunerated services. PMID- 17128730 TI - College member's EXTRA fellowship fuels change at The Capital Care Group. PMID- 17128731 TI - Evaluation of planning and implementation for an innovative health centre. AB - The South Calgary Health Centre, a new venture for the Calgary Health Region, was mandated to be an accessible, integrated, community-based health services facility. A comprehensive evaluation provided details on planning and implementation. Functional integration was a large focus of the planning and was operationalized as central admitting, shared charts, shared space and co-location of clinical services with no history of integration. Challenges are discussed and recommendations for future health centres developed. PMID- 17128732 TI - Governance, public participation and accountability: to whom are regional health authorities accountable? AB - The discourse of health reform has emphasized the need to increase public participation in decision-making as a way to enhance accountability. Despite recent gains in public participation in health care--primarily through citizen governance of regional health authorities--a clearly articulated accountability framework remains elusive in the Canadian context. If citizen participation is to be effective and meaningful, governing boards and politicians need to adopt an accountability framework that clearly delineates roles and responsibilities. PMID- 17128733 TI - Marginalized evidence: effective knowledge translation strategies for low awareness issues. AB - There are particular knowledge translation (KT) challenges in communicating evidence on low awareness topics. Often considered peripheral to the main focus of health care provision, these issues may fail to "get on the agenda" of decision-makers, even when supported by strong evidence. This article describes the successful application of KT strategies to a specific low awareness issue (the importance of using trained health interpreters) within a large Canadian Regional Health Authority. PMID- 17128734 TI - Assessing satisfaction with care in long-term care homes: current and best practices. AB - Given the increased call for accountability and quality of care in long-term care, it is important that government and long-term care provider organizations contemplate resident and family satisfaction with long-term care. This article highlights important considerations and provides practical recommendations for conducting satisfaction surveys in long-term care homes in terms of content, method of implementation and analysis, and use of findings. We conclude by recommending three surveys worthy of consideration by the government and long term care provider organizations in Canada. PMID- 17128735 TI - The fusion of PACS and RIS. PMID- 17128736 TI - The multiple facets of PACS: case history. PMID- 17128737 TI - Power to the people. PMID- 17128738 TI - Bridging the gap. PMID- 17128739 TI - Sustain the gain: case history. PMID- 17128740 TI - An enterprising answer: case history. PMID- 17128741 TI - Receiving is believing. PMID- 17128742 TI - Banking on safety. PMID- 17128743 TI - On the road to the EHR, keep sight of the legal health record. PMID- 17128744 TI - American's best health plans. PMID- 17128745 TI - Plans that won't report. PMID- 17128746 TI - The 2006 rankings. PMID- 17128747 TI - Stretching Medicaid. PMID- 17128748 TI - Careful consideration. PMID- 17128749 TI - Standardized chaos. PMID- 17128750 TI - The art of listening. PMID- 17128752 TI - JTMA helps boost diversity. PMID- 17128751 TI - Physician staffing in emergency departments: results of a Texas survey. PMID- 17128753 TI - Cultural humility in medicine: beyond the K-20 pipeline. PMID- 17128754 TI - Fee frenzy. PMID- 17128755 TI - Family funding fight. PMID- 17128756 TI - Clip, don't nick. AB - TMA and others have joined the 10,000 Lives Campaign to reduce patient deaths and injuries in hospitals. Part of the effort is reducing the risk of infection by using clippers instead of razors to shave tients before surgery. TMA also has specific recommendations n including keeping track of medications, for improving out-comes Ior Texas patients. PMID- 17128757 TI - Keep your guard up. AB - Pardon us if you've heard this before, but you really need to keep a close eye on the HMOs that settled the antiracketeering lawsuits against them to make sure they comply with terms of the settlement. PMID- 17128758 TI - Pitfalls and joy: my life as a doctor. PMID- 17128759 TI - High school athletic departments as sentinel surveillance sites for community associated methicillin-resistant staphylococcal infections. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is an emerging infection in athletes. Our study assessed MRSA burden in Texas 4A and 5A high school athletic departments by contacting 447 licensed athletic trainers (LATs) regarding skin infections in athletes; 186 (41.6%) responded. Sixty LATs reported MRSA in their athletic departments. The largest MRSA outbreak (23 infected persons) occurred in football players. The trainers also reported MRSA in wrestlers, volleyball players, cross-country runners, nonathlete students, and adults. Students and adults involved in high school athletics require MRSA intervention because of their large numbers and extensive contacts. Physicians should be aware of the potential for MRSA and should culture rather than treat empirically, communicate with school health staff to maximize surveillance for affected students when MRSA occurs in their student community, and contact their health department when the number of students with MRSA meets the unusual group expression, outbreak, or unusual severity criteria. PMID- 17128760 TI - Disasters bring out the best. PMID- 17128761 TI - Our influenza policy needs a cure. PMID- 17128762 TI - Consequences of living. PMID- 17128763 TI - Access granted. PMID- 17128764 TI - Guide for herbal product use by Mexican Americans in the largest Texas-Mexico border community. AB - Use of herbal products is a very common practice among Mexican Americans living in the El Paso, Texas, region. Although medicinal plants are generally assumed to be a safe and natural alternative to conventional medications, some herbal products could pose a potential health risk to the consumer. Currently, only a few studies focus on herbal product use by Mexican Americans, and no studies have focused on its use by patients with human immunodeficiency virus (HN) living on the US-Mexico border. A prospective observational study was conducted in 439 non HIV patients, as well as in 35 patients suffering from HIV Findings showed that 347 out of 439 (79%) of non-HIV patients and 25 out of 35 (71%) of HIV patients reported using herbal products. PMID- 17128765 TI - [Analysis of different constitutional, clinical, radiographic factors of 160 patients with low back pain based on own studies, observations and literature]. AB - BACKGROUND: Low back pain has both different etiopatogenesis and clinical symptoms. The most common reason of pain is degeneration of intervertebral disc. Development of civilisation causes an increase in prevalence and treatment's costs. The aim of this study was to: (1) An assesment of different clinical and radiographic factors amonr patients with low back pain; (2) A comparision of autors own observations with literature. MATERIAL AND METHODS: A group of 160 patients with clinical symptoms was analysed. Two groups A and B were distinguished. Group A consisted of 105 persons with centralisation of symptoms, while group B consisted of 55 persons with peripheralisation of pain. The analysis and assesment of several basic and constitutional factors and as well X rays and MRI findings was done. Statistical analysis was conducted in order to compare a frequency of occurence of studied factors. RESULTS: Results from material analysis and used methods were presented in final conclusions: (1) Centralisation of symptoms was connected with dimishing a pressure on nerve roots; (2) Statistical analysis revealed that among patients with peripheralisation, lateral shift, Lasegue's sign, and MRI were of significant value comparing to patients with centralisation; (3) Psychological problems, compensation demands, and waiting list phenomenon should be taken into consideretion in an assesment of low bac pain. PMID- 17128766 TI - [In vitro assessment of the lumbo-sacral spine after posterior transpedicular stabilization and anterior cage implantation]. AB - The biomechanical tests were performed on 9 cadaver specimens of lumbo-sacral spine. The specimens consisted of two lumbar motion segments, sacral bone together with the sacroiliac joint and parts of pelvis. The goal of the biomechanical tests was to estimate the motion of the lumbar spine, which was stabilised by the transpedicular implant and by the intervertebral cages. The tests showed that the traspedicular stabilisation is more effective than inervetebral with cages. PMID- 17128767 TI - [Anterior and posterior stabilization of the lumbosacral spine with the usage of interbody cages in the operational treatment of the isthmic spondylolisthesis]. AB - In the following work results of the operational treatment of the isthmic spondylolisthesis by the posterior stabilization and anterior lumbosacral interbody fusion with the use of interbody implants--cages was taken under evaluation. The test group consisted of 21 patients (13 male and 8 male). The follow up period exceeded 2 years. The objective clinical outcome assessment was based on Oswestry disability questionnaire. Subjective clinical evaluation was done by the visual analog pain score and two questions concerning the evaluation of success of the operative treatment and a possible agreement to a following operation if necessary. The radiological results were done upon evaluation of the degree of the spondylolisthesis, the angle of the lumbosacral lordosis, the height of the interbody space and intervertebral foramen and the evaluation of the spinal fusion. The conclusion was that the usage of the distraction of the lumbosacral spine in the operational treatment of the isthmic spondylolisthesis result in the reduction of the slippage and the dynamic decompression of the compressed neural roots. The usage of the interbody cages prevented the loss of slippage correction, permanently reconstructed the anatomical conditions in the area of the operated spinal segment and helped to achieve good and very good clinical results in over 95% of patients. The fusion rate was 100%. The restoration of the correct height of the intervertebral foramen in the slip segment caused an improvement of the neurologic state. The usage of two level stabilization in the operative treatment of the isthmic spondylolisthesis prevented the initiation of the secondary degenerative changes adjacent to the fusion. PMID- 17128768 TI - [Luxation of the prothesis after total hip arthroplasty]. AB - In this report we evaluated the incidence of complications following the total hip arthroplasty. We have focused on the role of the surgical approach as thepatient cause of the luxation of the prosthesis. Retrospective analysis of 393 endoprosthetic surgeries performed in the Department of Orthopedics and Traumatology, Pomeranian Medical University, between April 2002 and December 2003 was conducted. Out of 9 patients with prosthesis luxation 6 patients were operated using the posterior approach. A comparative analysis of selected groups did not reveal any significant differences in clinical condition of the patient, the implantation technique or the type of the applied prosthesis. We suggest that the main cause of prosthesis luxation were mistakes that led to misaligment of the prosthetic elements, and the posterior approach was believed to be the the predisposing factor. PMID- 17128769 TI - [Old and new materials in the hip prosthesis]. AB - Since the second half of the XX century a steady evolution of hip prosthesis design is taking place. Implant shape, fixation systems and materials are constantly evolving. The paper presents the materials combinations most commonly used in hip prosthesis (metal-metal, metal-polyethylene and ceramic). All the above mentioned materials have been in used for over 25 years, and thanks to minor modification and better quality are widely used all over the world. The authors basing on biochemical studies, clinical observations and personal experience present problems related to materials used in hip prosthesis. PMID- 17128770 TI - [Femoral head resection in dislocated hip in cerebral palsy children]. AB - The purpose of this study is to assess the outcome of proximal femoral resection in dislocated hip in cerebral palsy patients. Between 1993-2003 we treated 5 nonambulatory patients with quadriplegic form of cerebral palsy. We performed 6 resection of the proximal part of the femur. The average age at the surgery was 15.8 years (9-19.5 years) and average follow-up was 69 months (12-144 months). The indication to surgery was persistent hip pain. The results were good--the pain relieved, the movement in ,,the hip" was better and hygiene improved. Femoral head resection is radical, invasive method but we recommended it for selected group of severly spastic nonambulatory cerebral palsy patients with painful hip dislocation. PMID- 17128771 TI - [Hip joint development after closed reduction, complicated by growth disturbance of the capital femoral epiphysis]. AB - The purpose of the study was to assess hip joint development after the treatment of DDH with overhead traction and closed reduction complicated by growth disturbances of the proximal femoral epiphysis. Comparison was also made between joints affected by AVN and other treated joints. Antero-posterior radiographs of 107 hip joints (81 children) done during treatment and follow-up were retrospectively reviewed. The average age was 14.2 months (from 5 to 33 months) and age at the final follow up 20.7 years (minimum 14 years). AVN was detected in 31 (29%) of the hip joints. Type II necrosis according to Bucholz and Ogden classification system was the most common one. In the hips affected by growth disturbances of proximal femoral epiphysis there were worse final clinical and radiological results, osteoarthrosis and elongation of acetabulum were more often seen in comparison to other treated joints. There was no difference in terms of other parameters describing acetabulum. In 19 out of 31 cases deformity of femoral head was noted. AVN may present as a wide spectrum of pathology from mild, probably not significant growth disturbances of physis to severe deformity and lateralisation of femoral head. Statistically hips with AVN had worse final results and in cases with good congruity prognosis is often uncertain due to aspherical femoral head and secondary osteoarthrosis. Acetabular development is most often unaffected by AVN. PMID- 17128772 TI - [Iatrogenic lesions of the lower extremity nerves--causes, prevention and indications to the surgical treatment]. AB - Iatrogenic lesions of the nerve might be of a deliberate or unintentional character. The clinical material consisted of 73 patients treated surgically with unintentional injuries of the lower limbs nerves. Based on this material we analysed the causes of iatrogenic lesions of the lower limbs nerves and the probably mechanism of injury. The methods of prevention of these complications and the indication to surgical treatment were also described. PMID- 17128773 TI - [Neer classification of the proximal humerus fractures based on our own material]. AB - INTRODUCTION: About 4-5% of all fractures are localised to the proximal end of the humerus. During the last 30 years a two-time increase has been observed. The cause was ageing of population and a higher incidence of fractures. Treatment improved owing to proper classification based on radiological diagnostics. OBJECTIVE: The principles of diagnostic and classification were presented on the basis of the literature and analysis of the clinical material. MATERIAL: We analysed the ways of the diagnosis and classification of the proximal humerus fractures in 166 patients treated in Traumatologic and Orthopaedics Department of the Bielanski Hospital in Warsaw and in Orthopaedics Department of the Medical Postgraduate Medical Education Centre in Otwock during years 1988-2004. RESULTS: The classification suggested in 1970 by Neer has firmly established the way of treatment, is easy to remember, and regarded as the most popular and acceptable practice. It is based on dislocation one or more of the four bone fragments: shaft, head with articular surface, greater and lesser tuberosity of the humerus. Proper assessment of the type of fracture requires precise identification of these elements on radiograms. Correct assessment of dislocations of bone fragments and the way of dislocations is essential matter because specificity of the blood supply that improves heeling of the fracture and efficiency of the affected limb in the future. SUMMARY: It was found that conventional plain radiograms made in AP and Y exposures were sufficient for recognition the type of fracture. When diagnosis was questionable examination had to be broadened by axillar exposure and CT performed in the stated sequence. PMID- 17128774 TI - [Sacral pressure sores and their treatment]. AB - INTRODUCTION: Sacral bed sores still present a serious problem in most surgery departments. They occur mainly in elderly patients of limited mobility. The treatment of such sores extends over long periods of time and therefore involves considerable costs. MATERIAL AND METHODS: The material consisted of 11 sacral pressure ulcers treated surgically. The sores occurred in 4 severely disabled patients suffering from proximal third femur fractures, 4 patients with traumatic brain injury (treated in the Intensive Care Unit), and 3 patients suffering from bed sores after spinal cord injury. In 6 patients a fasciocutaneous flap was applied to the sores and in 5 cases a pedicled musculocutaneous gluteus maximus flap. The end results were assessed using Seiler's criteria. RESULTS: Complications of the "seroma" type were observed in 3 patients, and in 2 marginal necrosis. In all our patients complete healing was achieved within 2-4 weeks. CONCLUSIONS: On analysing our experience to date in surgical treatment of bed sores we are of the opinion that even extensive sacral sores can be covered with unilateral pedicled flaps provided that they are appropriately planned. Deep sores of the 4th degree sometimes with concomitant osteomyelitis require pedicled muscle flaps or in some cases musculocutaneous flaps to improve local circulation. The preparation of the patient for reconstruction surgery is just as important as the operation itself and therefore such preparation should never be neglected. PMID- 17128775 TI - [Ehlers-Danlos syndrome with concomitant joint contractures]. AB - We present 6 children with Ehlers-Danlos syndrome (EDS) with concomitant joint contractures. In 3 of them bilateral clubfeet together with hip, and (or) knee, or wrist flexion contractures were observed. In all of them, after initial soft tissue surgery for clubfeet, reccurences were noted requiring another surgery. Clubfeet were the only manifestation of joint contractures in another 2 children. In one of them soft tissue surgery was succesful, in the second a hypercorrection was noted. One child with EDS had a recurvatum knee deformity which was succesfully treated conservatively. We speculate on two different forms of EDS with joint contractures: a mild one, in which clubfeet are the only joint contractures, and in which a soft tissue surgery may lead to overcorrection, and a severe one, with multiple joint contractures like in arthrogryposis multpiplex congenita, in which soft tissue surgery for clubfeet and joint contractures may lead to reccurences. PMID- 17128776 TI - [Assessment of cytokines in the bone fracture union complications]. AB - The opinion on the usefulness of selected genetic examinations in connective tissue diseases is reported. Particular attention was paid to healing of fractures especially in respect of pathological aspects of accretion processes. The reasons for a delay in or a lack of bone fracture union process have not been fully explained--in spite of several reports relating to this type of examination. There are a number of egzogenic and endogenic agents including cytokines which regulate these processes. Up to date views on the suspected influence of cytokines on genetic polymorphism are discussed in present paper. Genetic polymorphism of some cytokines may increase the risk of post-menopausal osteoporosis. It seems that genetic factors are likely to cause complications of fracture healing. PMID- 17128777 TI - [Assessment of HBV, HCV and HIV injection in a population of Polish orthopedic surgeons]. AB - Orthopedic surgeons are at risk for occupationally acquired infections with blood borne pathogens. OBJECTIVE: To estimate the prevalence of infection with HBV, HCV, CMV and HIV among orthopedic surgeons. DESIGN: Voluntary, anonymous serosurvey at an annual meeting of Polish Association of Orthopedic Surgeons held in Szczecin, Poland in 2004. Serum samples were tested for anti-HIV, anti-CMV IgG, anti-HCV and markers of HBV infection: anti-HBc total and HBs. RESULTS: Of 1000 eligible orthopedic surgeons at the meeting, 101 (10.4%) participated; 75% participants reported a percutaneous blood contact in the previous month. None of the doctors was positive for HIV (0%, 95% CI:0-3.7%). One participant (1%, 95% CI: 0.2-5.4%), 26 years in profession, had anti-HCV. There was evi-dence of infection with HBV in 10 of 96 participants (10.4%) who had reported having no nonoccupational risk factors and in 5 participants with such factors. None of them developed a chronic infection. Only 5 out of 15 doctors infected with HBV knew their serological status, 13 out of those 15 had been immunized with hepatitis B vaccine, 4 revaccinated. The immunization rate was 91%. The seroprevalence for CMV was 63/101 (62%); it increased with age (p < 0.0003). CONCLUSIONS: Despite infection control precautions and availability of hepatitis B vaccine, orthopedic surgeons remain at risk for acquiring bloodborne viral infection. CMV poses the highest risk, followed by HBV and HCV. As the majority of HBV infected doctors did not know their serological status and underwent immunization with hepatitis B vaccine, testing for anti-HBc before vaccination remains crucial. PMID- 17128778 TI - [Metacarpal fractures--treatment results]. AB - In the study we presented the results of treatment of 126 fractures in patients hospitalized between 1994-2003 in the Department of Orthopeadics. The assesment was based on 87 patients that presented for the study, including 62 male and 25 female patients in the 14-57 age. The treatment consisted of stationary reduction combined with cast fixation and stabilization with Kirschner wire. The results were dividet into 5 groups based on the fractured metacarpal bone. The best results have observed the couse of treatment of 2nd, 3rd, 4th metacarpal fracturs and not so good in the 1st and 5th metacarpal fractures. PMID- 17128779 TI - [News in treatments of bone fractures on the beginning of XXI century]. PMID- 17128780 TI - [Medical-legal evaluation of peripheral chronic vascular disease]. AB - The prevalence of chronic obstructive vascular disease of the inferior limbs has been increasing in the western countries. As a consequence, the number of patients affected by this pathology, who undergo public health or forensic medicine examination to obtain the law-established benefits, is constantly rising. This paper illustrates the pathophysiology, epidemiology, clinical and prognostic factors used to attribute the Fontaine and Rutherford class, and the instrumental evaluation parameters necessary for a medical-legal assessment. The authors describe also the variety of welfare and social actions that the social security system provides to this type of patients in Italy, with an overview of damage assessment parameters within the forensic medicine. Finally, the authors suggest medical-legal criteria for the staging of permanent damage in these patients, based on stenosis degree, on systolic pressure measured at the ankle at rest and after exercise, on ankle brachial index, on Doppler morphology and spectral analysis of the velocity peak, and on the walking distance ability. PMID- 17128781 TI - [The pro-inflammatory role of cytokines in the mechanism of atherosclerosis]. AB - Investigation of the mechanisms of atherosclerosis has determined that inflammation plays a central role in the development, progression, and outcome of acute coronary syndrome. Although C-reactive protein will remain over time a useful marker, cytokines will continue to be studied in order to understand the mechanisms of acute coronary syndrome and cytokine balance. This short review summarizes the experimental and clinical evidence regarding the role of some cytokines in acute coronary syndrome (interleukin [IL]-8, IL-10, IL-18, IL-2, tumor necrosis factor-alpha, interferon-gamma). PMID- 17128782 TI - [Biochemical markers for predicting chemotherapy-induced cardiotoxicity: systematic review of the literature and recommendations for use]. AB - Chemotherapy is a well established therapeutic approach for several malignancies, but its clinical efficacy is often limited by related cardiotoxicity leading to cardiomyopathy evolving towards heart failure that may worsen the patient outcome. To detect cardiac damage, the most frequently adopted diagnostic approach is the estimation of left ventricular ejection fraction by echocardiography, showing, however, low sensitivity in early prediction of cardiomyopathy, when appropriate treatments could still improve the patient's outcome. Cardiospecific biomarkers, like cardiac troponins, show high diagnostic efficacy in the early, subclinical phase of disease, becoming positive approximately 3 months before clinical onset of cardiomyopathy. Furthermore, the increase in their concentrations is well correlated with the disease severity and may predict the occurrence of major cardiac events during follow-up. On the other hand, negative troponin concentrations may identify patients with a very low risk of cardiomyopathy (negative predictive value = 99%). For cardiac natriuretic peptides, definitive evidence about a diagnostic or prognostic role in predicting chemotherapy-induced cardiomyopathy is lacking and their practical use in this context cannot be recommended until their clinical efficacy is clearly defined. PMID- 17128783 TI - [Pacemaker implantation in children: is this a therapy or a pathogenetic mechanism for ventricular dysfunction?]. AB - Ventricular pacing, typically initiated from a right ventricular apical electrode, inherently causes abnormal biventricular activation, decreases left ventricular function, and may cause histopathological changes. Data regarding cardiac resynchronization therapy in pediatric patients are limited. Some authors evaluated the potential of improving paced left ventricular function by a septal electrode implant site. For this reason the role of pacing is not clear in children. In isolated congenital atrioventricular block, cardiac pacing represents a satisfactory treatment. However, recent evidence has demonstrated that a subset of patients with isolated congenital complete atrioventricular block develops dilated cardiomyopathy. Patients with congenital heart diseases after surgery represent a unique and expanding population. Some will require pacemaker or implantable defibrillator therapy. They pose technical and management challenges not encountered in other groups receiving pacing, and the complication and reintervention rates specific to this population are not well defined. Moreover, the small patient size, structural cardiac abnormalities, and growth may complicate pediatric pacemaker management. Better knowledge of risk factors for lead failure in these patients may help improve future outcomes. In this review we analyzed many of these problems. PMID- 17128784 TI - [Surgical therapy of coronary artery disease in the Italian cardiac surgery centers]. AB - BACKGROUND: We conducted a national survey on surgical therapy for coronary artery disease and its complications, with the aim to eventually identify differences in the processes of care adopted in the Italian cardiac surgery centers. METHODS: Eighty-seven Italian cardiac surgery units received a questionnaire about their general activity profile and the following interest areas: (a) surgical myocardial revascularization techniques; (b) ischemic mitral regurgitation treatment; (c) left ventricular reconstruction procedures; (d) mechanical circulatory support and heart transplantation in ischemic cardiomyopathy. RESULTS: Forty-one Centers (47.1%) returned a completely filled questionnaire. The analysis of the answers showed differences in the therapeutic approaches adopted in the various centers regarding: choice of surgical technique and grafts for myocardial revascularization; propensity to treat moderate ischemic mitral regurgitation in the setting of severe left ventricular dysfunction; propensity to treat extended myocardial scars; treatment options in ischemic cardiomyopathy with advanced heart failure. The specific center profile (volume of activity, availability of heart transplantation) seems to influence the choice among the different treatment options. CONCLUSIONS: This study may contribute to show and analyze existing differences in clinical practice between different centers. Diagnostic and therapeutic approaches (processes of care) should be frequently monitored in order to promptly single out critical areas necessitating review or further researches, to standardize decision making criteria, and to improve care appropriateness. PMID- 17128786 TI - [Of hubs, spokes and acute coronary syndromes]. PMID- 17128785 TI - [Transfer of patients with acute coronary syndromes]. AB - BACKGROUND: The creation of a network to treat patients with acute coronary syndrome with revascularization devices presents a series of critical issues, one of which is the transfer of this type of patients from a spoke to a hub hospital with angioplasty facilities. We investigated the means of transport and the adverse events that could occur during such transfers. METHODS: We analyzed data relating to patients moved from our spoke hospital to hubs over a period of 2 years, utilizing medical records and transport reports. RESULTS: During this period, 531 patients aged 30-91 years were moved; complete clinical data and transfer reports were available for 495 (93%). There were 42 patients with a diagnosis of ST-elevation myocardial infarction (STEMI) <24 h; 115 with STEMI >24 h; 33 with non-ST-elevation myocardial infarction (NSTEMI) <24 h; 156 with NSTEMI >24 h; 122 with unstable angina; and 27 with other diseases. Of these patients, 157 had angina in the 24 h before the transfer; 24 had signs of hemodynamic instability; and 18 had electrical instability, for a total of 166 "unstable patients"; 294 patients (59%) were moved in a medically equipped ambulance with a cardiologist and a professional nurse on board. There were three adverse events (< 1%) but no deaths: one cardiac arrest, one acute pulmonary edema, and one relapse of myocardial infarction. All 3 patients showed signs of clinical and hemodynamic instability at the time of transfer. CONCLUSIONS: The transfer from spoke to hub of patients with acute coronary syndrome who need coronary angiography can be done easily and safely, with a low incidence of adverse events, which are correlated with preexisting clinical and hemodynamic instability. PMID- 17128787 TI - [Therapeutic obstinacy in cardiology]. PMID- 17128788 TI - [The network configuration of healthcare organizations: an introduction to a novel sociological paradigm]. PMID- 17128789 TI - [Aortic valve replacement on two octogenarians awake patients without tracheal intubation, with thoracic epidural anesthesia]. AB - The association of advanced age with various comorbidities increases the risk of mortality and morbidity in cardiac surgery. The utilization of high thoracic epidural anesthesia (HTEA) in this setting presents numerous potential benefits, including early recovery of consciousness and of spontaneous ventilation, hemodynamic stability, enhanced analgesia, improved pulmonary function, and earlier recovery. Moreover, this anesthesiological technique allows the performance of surgical procedures on the conscious patient, thus making continuous monitoring of the cerebral function feasible. We have employed HTEA without tracheal intubation on 2 gravely compromised octogenarian patients who underwent aortic valve replacement for critical aortic stenosis. Epidural anesthesia without tracheal intubation in these patients permitted the avoidance of general anesthetics and allowed the continuous evaluation of their cognitive function. Further, by avoiding the positive pulmonary pressures of mechanical ventilators, the technique contributed to preserve physiologic intrapulmonary pressures, thus positively affecting the pulmonary circulation. In our opinion, the utilization of HTEA without tracheal intubation may decrease the surgical risk in selected patients. PMID- 17128790 TI - [The work of the Istituto Superiore di Sanita (ISS) on the short-term outcome of aortocoronary bypass in Italy]. PMID- 17128791 TI - [Urogenital tuberculosis: clinical presentation, diagnosis, and treatment]. PMID- 17128792 TI - [Criteria for prediction of recurrent sarcoidosis]. AB - The study based on history data, including age, educational level, social status, smoking, occupational risk factors, and a history of allergy and comorbidity, and on radiation diagnostic and fibroscopic findings could yield criteria for predicting potential recurrent sarcoidosis at the early stages of diagnosis of the pathological process. PMID- 17128793 TI - [The clinical and functional features of chronic obstructive lung disease in patients with organic chlorine pesticides in blood]. AB - Impact of pollutant human aggression is of great importance for the development of bronchopulmonary diseases. In the Republic of Uzbekistan, the environmental levels of pesticides are elevated as they are widely used in agriculture while growing cotton. Eighty-two patients with chronic obstructive lung disease (COLD) were studied in relation to the presence of blood chlorine- and phosphorus containing pesticides. The latter were tested for their effect on the course of COLD. There was evidence that the higher blood levels of pesticides were, the more severe the clinical symptoms of the process and the more pronounced changes in the functional status of the bronchopulmonary apparatus were. The findings suggest that the blood content of organic chlorine pesticides affects the course of COLD. These compounds cause more significant clinical and functional impairments with the severe manifestations of mucous membrane morphofunctional rearrangement and with disruption of the body's adaptive potentials. These patients poorly respond to therapy, which required dose adjustment and longer drug use. PMID- 17128794 TI - [Use of mathematical models for interpreting the tuberculosis epidemiological situation]. PMID- 17128795 TI - [Alcohol sales and pulmonary tuberculosis mortality in the Republic of Belarus in 1981 to 2001]. AB - The fact that there is an association of alcohol abuse with pulmonary tuberculosis is well documented. The effect of alcohol sales per capita on tuberculosis mortality rates is considerably less known. The aim of the study was to evaluate the beverage-specific effect of alcohol on pulmonary tuberculosis mortality rates. Trends in pulmonary tuberculosis mortality rates in Belarus from 1981 to 2001 were analyzed in relation to those in the level of sales of various alcoholic beverages per capita, by applying the time series analysis. The analysis demonstrated a positive and statistically significant effect of changes in per capita alcohol sale levels on pulmonary tuberculosis mortality rates. It suggests that a 1% increase in alcohol sales per capita might cause 0.49 and 0.36% increases in pulmonary tuberculosis mortality rates in males and females, respectively. This study also indicates that tuberculosis prevention programs should place more emphasis on alcohol problems. PMID- 17128796 TI - [Significance of risk groups in pediatric phthisiology]. AB - Detailed analysis of risk factors in 161 children with pulmonary tuberculosis has established that tuberculosis risk-group children are ill in most cases (82.6%). Examining the results of annual tuberculin diagnosis in 2478 children aged 1 to 14 years in a children's city polyclinic could ascertain that the most risk groups comprise children with a conversion of tuberculin tests (5.2%) and tuberculosis-infected children with increased tuberculin sensitivity (7%) who are the basic contingent of followed up local district pediatric phthisiologists. The priorities of activities of pediatric phthisiology involve the detection of tuberculosis risk groups by the general pediatric service by the tuberculin diagnostic technique under the guidance of a phthisiological service and the choice of individual management policy by their pediatric phthisiologists. PMID- 17128797 TI - [Genetic dependence of adaptive variability in Mycobacterium tuberculosis]. AB - The investigations have established that all the species of Mycobacterium tuberculosis formed in the course of evolution are the derivatives of a single causative agent with inherent properties of vital activity in human beings, various species of animals and poultry, which makes it possible to adapt them in the untypical host and to cause tuberculosis. The findings indicate that the etiological role of M. bovis and M. avium in human tuberculosis is significant, especially in children as the main consumers of dairy products and eggs from tuberculosis-infected animals and poultry. All the species of Mycobacterium tuberculosis are dangerous to the hosts uncharacteristic of them. PMID- 17128798 TI - [The severity of clinical signs of a tuberculous process in first detected patients]. AB - The severity of clinical signs of pulmonary tuberculosis was studied in first detected patients. A hundred and sixty-eight case histories of the patients admitted to a town tuberculosis dispensary in 2004 were analyzed. A total of 87 (51.8%) patients with tuberculosis-suspected complaints visited the health care facility. Active preventive fluorographic studies revealed 81 (48.2%) patients. A hundred and twelve (66.6%) patients were diagnosed as having severe destructive forms of the disease in the phase of decay and dissemination. The clinical signs of a tuberculous process were less significant or absent in actively detected patients. Later detection of patients makes their people contact them longer and increases the probability of development of an active tuberculous process in the latter. PMID- 17128799 TI - [Capacities of rapid diagnostic methods in renal tuberculosis]. AB - The paper comparatively assesses routine and new medical technologies (metachromatic urine test, polymerase chain reaction, chromatographic mass spectrometry, and enzyme immunoassay) for detection of Mycobacterium tuberculosis and tuberculosis antibodies. It is shown that molecular biological and immune assays may be used only for rapid diagnosis and screening, followed by a full scale phthisiourological study as the detection of markers of Mycobacterium tuberculosis and tuberculosis antibodies cannot be a criterion for establishing renal tuberculosis. The authors consider that the conventional microbiological tests whose validity is beyond question in case of positive results are the most established and practice-tested canons of phthisiourological diagnosis. PMID- 17128800 TI - [Use of essential oil of peppermint (Mentha piperita) in the complex treatment of patients with infiltrative pulmonary tuberculosis]. AB - The paper describes the effects of peppermint (Mentha piperita) essential oil inhaled by patients with infiltrative pulmonary tuberculosis in the penitentiary system. This procedure is shown to be most effective in infiltrative pulmonary tuberculosis in the phase of resorption of infiltrates and/or closure of decay cavities. The efficiency is determined by the rapid positive changes in a tuberculous process, which appear as a rapider regression of tuberculous inflammation, causing small residual changes. This procedure may be used to prevent recurrences and exacerbations of pulmonary tuberculosis. PMID- 17128801 TI - [Adverse neurotoxic reactions of chemotherapy for tuberculosis and their treatment]. AB - A hundred and sixty-five first detected patients with pulmonary tuberculosis, receiving routine chemotherapy regimens were followed up for 4 months. Neurotoxic reactions of the central and peripheral nervous systems, which required isoniasid discontinuation, were developed in 13.3 and 11.5% of the patients, respectively. Therapy for neurotoxic reactions used cortexin in the study group (n = 22) and gamma or vitamins B in combination with glutaminic acid in the control group (n = 19). Cortexin used in the therapy of neurotoxic reactions produced a beneficial effect and permitted 90.9% of the patients to continue to use isoniazid while 42.2% of the controls resumed using isoniazid. PMID- 17128802 TI - [A method for pre-seeding treatment of biomaterial for mycobacterial isolation]. AB - Ethonium solutions were used as a detergent for the pretreatment of a biomaterial homogenate to isolate mycobacteria. The best results were obtained when 1% ethonium solution was used for these purposes for 30 minutes. The pretreatment of a biomaterial homogenate in this mode increases the rate of mycobacterial isolation and reduces the time of tests. The method may be also effective for the pretreatment of human sputum when it is bacteriologically tested for mycobacterial isolation. PMID- 17128803 TI - [Bronchial obstruction in complicated pulmonary hydatid disease: methods of its correction at the stage of preoperative preparation]. PMID- 17128804 TI - [Determining the antibiotic resistance profile of the causative agents of secondary (mixed) infections in pulmonary tuberculosis]. PMID- 17128805 TI - The skin notation in the MAC list and classification of dangerous chemicals. AB - The European Union has published a list of dangerous chemicals, along with classification and labeling; in 1997 the list was adapted to the requirements of Polish regulations and has been continuously updated since then. We have decided to adopt data on dangerous chemicals classification in the list referring to their dermal absorption to assess whether the skin notation is required in the maximum admissible concentration (MAC) list. In Poland, the Group of Experts for Chemical Agents (GECA) decides on assigning the skin absorption notation (symbol Sk) when analyzing the literature data collected in order to prepare occupational exposure limits documentation. The LD50s value serves as the main criterion for assigning the notation. The limit value of 1000 mg/kg has been set as the criterion for applying the skin notation. Documented results of animal and human research, which point to systemic effects resulting from dermal exposure and physicochemical characteristics enabling calculation of the dermal absorption rate may also be used as the basis for assigning the skin notation. Chemicals for which GECA recommended using the skin notation in the 2004 MAC list have been analyzed. It was concluded that information on the classification of chemicals in the list of dangerous chemicals is useful for assessing the skin absorption, but quantitative assessment of absorption rate compared to "safe" levels, using the available theoretical models, should also be attempted. PMID- 17128806 TI - New approach to environmental tobacco smoke exposure and its relation to reemission processes. AB - Indoor air quality is one of the factors that determine human well-being and health. Being aware of this fact, it is essential to identify the origin, kind, mechanism, and effects of harmful substances contained in the air. The issue concerning the contents and primary emission of these substances from building materials and interior furnishings is well known. Adverse effects of environmental tobacco smoke (ETS), including exposure of passive smokers, are also very well documented. To the contrary, reports on secondary and indirect emissions, especially those focused on mechanisms by which pollution is "transferred" by materials used in interior furnishings are very rare. Textiles are used in a great variety of ways as functional and decorative materials. These materials in general, and textile floor coverings in particular, are extensively utilized in fitting apartments, public buildings, and transport means. Studies on this aspect of the role played by textile materials in ETS exposure have been only fragmentary documented. PMID- 17128807 TI - Occupational diseases in the period of socioeconomic transition in Poland. AB - OBJECTIVES: Current situation in the epidemiology of occupational diseases is a resultant of many contributing factors, such as occupational exposures of the working population, social and economic conditions, medical measures, legislation, and ongoing changes in the national economy. MATERIALS AND METHODS: This work is based on the information compiled from reporting forms on occupational diseases gathered in the Central Register of Occupational Diseases run by the Nofer Institute of Occupational Medicine, Lodz, Poland. RESULTS: A systematic downward trend in the number of occupational diseases has already been observed over several years. In 1998, as many as 12 017 cases were recorded (117.3 per 100,000 workers), while in 2005, this number was reduced to 3249 (34.8 per 100 000). In 2005, the highest incidence rates were noted for chronic voice disorders, pneumoconioses, infectious and parasitic diseases, hearing loss, and skin diseases. The observed decrease in occupational pathology is associated with continued transformations in the national economy. Changes in the nature and level of occupational exposures have influenced the profile of occupational pathologies. Medical preventive measures, such as vaccinations covering high-risk groups have contributed to the reduced incidence of occupational diseases. For example, the number of cases of occupation-related viral hepatitis was seven times lower in 2005 than in 1993. The 2005 data show that over 50% of occupational diseases were diagnosed in workers employed outside industry. CONCLUSIONS: Changes taking place on the labor market, new technologies and most of all transfer of the workforce from industry to the service and administration sectors, will certainly affect the profile of occupational and work-related pathologies. In the present-day environment, the work-related stress has become a dominant factor. PMID- 17128808 TI - Reducing health risks arising from child and adolescent labor. AB - OBJECTIVES: The aim of the studywas to work out a program that would help develop the most effective methods for protecting the safety and health of young workers. MATERIALS AND METHODS: The program was designed by a multidisciplinary team of experts in occupational and public health, occupational hygiene, health promotion, sociology, and regulatory and control activities. RESULTS: The program points out the major aspects to be considered while implementing the existing legislation. The employers should have a proper recognition of health hazards related to working conditions at their enterprises. These should be considered while assessing work capability of adolescents during the pre-placement and periodic medical examinations. Prophylactic examinations for adolescent workers should be oriented towards the potential or existing health hazards and strenuous conditions of in-plant vocational training. It is essential to inform them about work-related health hazards and the preventive methods to be applied by both the workers themselves and the occupational health services. If a health problem or ailment is detected by an occupational physician, the adolescent worker should be referred to primary health care unit for further diagnostics. CONCLUSIONS: The most effective ways to protect the safety and health of young workers include a more accurate assessment of working conditions, contracting prophylactic examinations with only one occupational health service unit, providing appropriate risk information to adolescent workers/students, and a closer collaboration between school administration, employers and occupational health services.. PMID- 17128809 TI - Occupational stress and its consequences in healthcare professionals: the role of type D personality. AB - OBJECTIVES: The purpose of the study was to investigate the role of Type D personality in perceiving stress at work and the development of adverse effects of experienced stress, i.e. mental health disorders and burnout syndrome. MATERIALS AND METHODS: A sample of 79 healthcare professionals (51 psychiatrists and 28 nurses) of a mental hospital in Lodz was eligible for the study. The mean age of the subjects was 39.71 (SD = 8.02) and their work experience was 11.20 (SD = 5.45). The DS-14 self-report to measure Type D personality, the Subjective Work Evaluation Questionnaire, the Maslach Burnout Inventory, and the General Health Questionnaire (GHQ-28) were used in the study. RESULTS: The results of the study confirm a significant role of Type D personality in perceiving job stress and the development of its adverse effects reflected especially in the worsening health condition. Type D subjects perceive their workplace as more stressful than non Type Ds and manifest more symptoms of mental health disorders and a higher level of burnout, expressed mainly in the form of emotional exhaustion and lower personal accomplishment. CONCLUSION: Modification of Type D personality aimed at reducing tendency to experience negative emotions and enhancing skills to express them combined with improving social relations is desired to prevent healthcare professionals from adverse health outcomes. PMID- 17128810 TI - Physiological reaction to work in cold microclimate. AB - OBJECTIVES: In Poland, occupational exposure to cold microclimate is quite common (5.1 workers/1000 occupationally active people). Reports on health effects of this exposure are rather scarce. The aim of the study was to evaluate the physiological reaction in workers occupationally exposed to cold microclimate. MATERIALS AND METHODS: Examinations were performed in a group of 102 workers (41 women and 61 men) employed at cold storage units. The mean age in the group was 39.1 +/- 9.9 years and the duration of employment under conditions of cold environment was over 12 years. The study population was divided into four groups, according to microclimate conditions (group I, ambient temperature -26 degrees C; group II, 10-14 degrees C; group III, 18-20 degrees C, control group; and group IV, 0-10 degress C). The workers underwent the following procedures: general medical examinations, cold pressor test, ambulatory blood pressure monitoring, and heart rate variability (HRV) analysis (time- and frequency-domain parameters). RESULTS: The results were adjusted for confounding factors (age, smoking and drinking habits). The analysis of HRV parameters did not reveal any significant differences between the study groups. However, systolic and diastolic blood pressure (BP) in the daytime and at night was significantly higher in group IV compared to group II. Mean heart rate (HR) in the daytime and at night and the BP and HR day/night ratio did not differ between the groups. The analysis of BP by gender revealed that in women, systolic BP during the day and at night was significantly higher in group IV than in group II. In the group of workers with hypertension (18 men and 5 women), men reacted to the cold pressor test either by increased or decreased BP while all the women reacted by the increased BP. CONCLUSIONS: Our findings indicated that in workers exposed to cold microclimate, the physiological reaction was dependent on gender and ambient temperature. Women seemed to be more sensitive to cold stress than men. However, this finding must be further investigated. PMID- 17128811 TI - Exposure to chlorphenvinphos, an organophosphate insecticide, prevents from behavioral sensitization to amphetamine. AB - OBJECTIVES: Exposure to organophosphorus (OP) pesticides, irreversible inhibitors of acetylcholinesterase (AChE), may result in long-lasting alterations in the functional state of the central nervous system. In earlier studies, we found that a single exposure of the rat to chlorphenvinphos (CVP), an OP pesticide, made the animal hyposensitive to amphetamine (AMPH) three weeks posttreatment. A repeated administration of AMPH is known to result in a progressive increase in the behavioral sensitivity to the psychostimulant. It makes it likely that treatment with AMPH after the CVP exposure may result in amelioration of the CVP-induced hyposensitivity to the psychostymulant. The purpose of the present experiment was to check out this supposition. MATERIALS AND METHODS: At the first stage, the relationship between the CVP dose and the effect on sensitivity to AMPH was tested. The rats were given CVP once intraperitoneally (i.p.) at a dose of 0.0, 1.0 or 3.0 mg/kg. Three weeks later their open field behavior was assessed before and after i.p. administration of 0.25, 0.5 or 1.0 mg/kg of AMPH. At the subsequent stage, the susceptibility of the CVP-treated rats to AMPH sensitization by repeated AMPH treatment was investigated. For this purpose each of the rats was repeatedly treated with AMPH in its home cage (one injection/day for five days). At stage two, the daily AMPH dose received by each animal was of the same magnitude as that received at stage one. Two weeks after the last AMPH treatment dose, the motor response to a test AMPH dose (0.5 mg/kg) was measured in all rats. RESULTS: The results of stage one confirmed a significant reduction of behavioral sensitivity to AMPH in the CVP-treated rats. The results of stage two indicated that the CVP-induced decrease in sensitivity to AMPH was not ameliorated by a repeated treatment with AMPH at any of the used doses. In fact, in the rats exposed to the high CVP dose, repeated treatment with AMPH resulted, dose dependently, in augmenting of the depressive effect of the pesticide. CONCLUSIONS: It appears then that treatment to an OP pesticide reduces the rat's sensitivity to AMPH and makes the animal resistant to sensitization by repeated treatment with the psychostimulant. PMID- 17128812 TI - Tissue distribution and excretion of N-methyl-2-pyrrolidone in male and female rats. AB - OBJECTIVES: N-methyl-2-pyrrolidone (NMP) belongs to solvents widely used in the petrochemical industry a well as in the production of pesticides, veterinary drugs and paint removers. NMP is easily absorbed from the respiratory tract, digestive system and through the skin. It is a compound of slight acute toxicity that also displays moderate irritating activity. The aim of this study was to assess tissue distribution and excretion following a single intraperitoneal NMP administration. MATERIALS AND METHODS: Tissue distribution and excretion of NMP following administration of a single dose of 250 mg/kg body weight (350 kBq/rat) was investigated using 14C. Blood plasma (6 rats per time point) were sampled up to 72 h after administration and determination of radioactivity. Male and female rats (4 animals per time point) were decapitated at appropriate time intervals and examined tissues were removed for determination of radioactivity. Excretion of 14C in urine and feces were also measured. All radioactivity measurements were carried out using a Rackbetta 1209 (LKB, Sweden) liquid scintillation counter. RESULTS: The highest 14C activity in tissues and internal organs of female and male rats was observed 4 h after administration of the compound. The highest accumulation was detected in the muscles and fat tissue as well as in the liver and testicles. During 72 h following administration, approximately 80% of the dose was excreted in urine. Elimination of the compound in feces was far less significant: only about 5% of the dose was excreted at once. CONCLUSIONS: The results of the study indicate that there are no significant differences in 14C NMP tissue distribution between male and female rats; NMP absorption from the peritoneal cavity to blood is rapid, disappearance from plasma is monophase and kidneys are the main route of excretion of NMP and/or its metabolites from the rat body after administration of a dose equal to 10% of LD50. The ability to accumulate NMP and/or its metabolites in testes and seminal vesicles may be the reason for fertility impairment in male rats observed after repeated exposure to this compound. PMID- 17128813 TI - Message from the Editor-in-Chief. Expression of concern due to plagiarism. PMID- 17128814 TI - [Association between socioeconomic status and oral hygiene among preschoolers enrolled in the IMSS preventive dental program in Campeche]. AB - OBJECTIVE: Determine the association between socioeconomic status and oral hygiene in the primary dentition of preschool children. MATERIALS AND METHODS: We undertook a cross-sectional study of 1,303 children attending 10 schools in Campeche, Mexico. Every child was clinically examined in a portable dental chair by one of four examiners. We used a questionnaire addressed to the mothers to collect data on socioeconomic and socio demographic variables--including attitudinal variables dealing with the perceived importance of oral health. Oral hygiene was assessed appraising the frequency of tooth brushing and the presence of dental plaque. Data analysis included non-parametric tests using STATA 8.2. RESULTS: Mean age was 4.36 +/- 0.79 years and 48.3% of children were girls. Of the study population, 17.8% (n = 232) were classified as having inadequate oral hygiene, 50.9% (n = 663) having moderate oral hygiene, and 31.3% (n = 408) having adequate oral hygiene. Children who were rated more frequently as having inadequate hygiene (p < 0.05) had mothers with a negative attitude toward oral health, were users only of public medical insurance (as opposed to users of private services), and had not used dental services in the year prior to the study. Finally, we observed a decrease in the adequacy of oral hygiene associated with a decrease in socioeconomic status. CONCLUSIONS: Our findings showed that oral hygiene was closely associated with socioeconomic status. This implies that if a reduction of oral health inequalities is to be achieved, the strategies and resources targeting these goals must take into account the existing differences between population groups with more or fewer social disadvantages. PMID- 17128815 TI - [Recombinant factor VII activated in non-hemophiliac patients with acute severe hemorrhage in the intensive care unit]. AB - BACKGROUND: Severe hemorrhage is a frequent complication with multiple etiologies and high morbi-mortality observed among critically ill patients. Recombinant Factor VIla (rFVlla) constitutes a new therapeutic alternative. OBJECTIVE: Analyze the evolution in a non-hemophiliac patient group with severe hemorrhage treated with rFVlla. MATERIAL AND METHODS: Ten non-hemophiliac patients with severe hemorrhage, five men and five women between 18 and 74 years, were included and treated with rFVIIa. We used a t test for statistic analyses. Significance was set at p < 0.001. RESULTS: Among patients treated with rFVlla, hemorrhage was controlled and the use of blood products was significantly diminished (p < 0.001). Coagulation tests, fibrinogen levels, platelet count and perfusion parameters increased significantly (p < 0.001). CONCLUSIONS: rFVIIa is a therapeutic alternative for the treatment of severe hemorrhage not controlled by conventional management. PMID- 17128816 TI - [Sporotrichosis among rural communities in the Northern Sierra in Puebla. Report of 55 cases September 1995 - December 2005]. AB - Fifty five cases of cutaneous sporotrichosis collected from 35 communities located in the southeast region of the Northern Sierra of Puebla are described. The disease was more prevalent in males (60:40), but this difference was restricted to younger subjects (aged 0-15 years) where 14 cases were male and only 3 were female. No statistical difference regarding gender was observed in elder patients. The prevalence of the disease was significantly lower among patients aged 31 to 45. The most common clinical forms of the disease were lymphocutaneous and fixed. The drug of choice for the treatment of patients in rural communities was potassium iodide. When available, Itraconazol proved to be an excellent option. PMID- 17128817 TI - [Micosis among five highly underprivileged Mexican communities]. AB - BACKGROUND: In many small rural communities in Mexico, medical care is deficient, empirical or absent. OBJECTIVE: In order to improve health coverage in rural areas, the Mexican Institute of Social Security organizes Medical and Surgical Meetings of various specialties including Dermatology and Mycology (MSDM). These include visits to rural hospitals by dermatologists and a mycologist to care for underprivileged communities. In addition to taking samples, they establish the clinical diagnosis and indicate medical and/or surgical treatment, with follow-up visits when needed. MATERIAL AND METHODS: In 2004 and 2005, five MSDM in Chiapas (two), Puebla (one), Michoacan (one) and Oaxaca (one) were organized. Mycoses were within the first four skin pathologies detected. RESULTS: Direct examination with potassium hydroxide led to the diagnosis of mycosis and other skin diseases such as scabies, pediculosis or hair disorders. The sample cultures showed, in addition to common fungi as dermatophytes (Trichophyton rubrum, 19 cases), other uncommon fungal agents such as Trichosporon spp, Chrysosporium spp, Cryptococcus, Geotrichum spp and Aspergillus spp. Most of the candidiasis cases were caused by Candida parapsilosis (nine cases) followed by C. albicans (three cases). PMID- 17128818 TI - [Longitudinal study of three families with familial Parkinson's disease]. AB - BACKGROUND: Familial Parkinson's is a variant of Parkinson's disease (PD) transmitted generationally with an early onset. OBJECTIVE: Describe the clinical disease characteristics and its 18 year evolution among families in Colima presenting familial PD. MATERIALS AND METHODS: We determined disease diagnosis, evolution and hereditary pattern. The UPDRS system was used to follow the longitudinal course of the disease. Descriptive statistics were carried out using means and percentages. RESULTS: Three families were studied, with a total of 51 subjects aged 29 +/- 22 years spanning 4 generations. Thirty-seven percent of studied subjects displayed familial PD, with disease onset at 24 +/- 9 years of age. The highest UPDRS value was 175. Disease transmission with a dominant autosomic heredity pattern was shown. One hundred percent of first and second generation members from family number 1 displayed the disease. CONCLUSIONS: The three families displayed early onset PD and rapid progression, coinciding with described characteristics of type 1 familial Parkinsonism (PARK1). This disease is caused by the Ala53Thr mutation of the alpha-synuclein gene. PMID- 17128819 TI - [Ten year experience with ocular lymphomas and ocular adnexae lymphomas]. AB - BACKGROUND: Ocular lymphomas and ocular adnexae lymphomas (OL and OAL) constitute 7-8% of all extranodal lymphomas. OBJECTIVE: Describe the clinical, morphologic and immunophenotypic characteristics of OAL seen in our hospital. MATERIAL AND METHODS: Retrospective analysis of patient records with OL and OAL between July 1994 and July 2005. The following data was analyzed: Clinical presentation, therapy, treatment response, overall survival and disease free survival. RESULTS: Ten patients with OL and OAL were identified. Of these, 8 were women and 2 men. Median age was 50. Eight of 10 patients achieved complete remission, 6 of the 6 presenting MALT Lymphoma. Two patients with stage IV had refractory disease. CONCLUSIONS: In our series 0.02% of lymphomas were OL and OAL of a total 498 LNH. MALT lymphomas appear at a more advanced age, sixty percent of the cases were MALT lymphomas and were diagnosed during their early stages. Patients were followed during 21 months, global survival was 100%, free illness survival had a mean of 868 days and a survival median of 442 days. PMID- 17128820 TI - [Physiopathology of prion diseases]. AB - Prion diseases are a group of degenerative disorders characterized by being progressive, fast growing, and fatal, they affect humans and animals. Due to their physiopathogeny, these disorders can be sporadic, genetic, or infectious. Prions are cellular proteins that lack nucleic acids; they are not viruses or microorganisms. Prions induce neuronal death, brain spongiosis, which are a hallmark of these diseases, as well as amyloid prion protein plaque aggregates. Although the causes that favor pathogenic prion proteins remain uncertain, it is possible that conformational changes of the prion protein allow them to create copies of themselves to form aggregates and induce neuronal death. Other theories suggest that quantitative and qualitative changes in the glycosylation pattern induce the pathological prion form. The latter allows to explain some of their interactions and to understand better the conformational changes and the physico chemical properties of the prion protein. We review some of the first biological functions (as a transporter of Cu2+ ions) that have been described to this molecule. The present review focuses on different aspects of prion diseases aimed at understanding better their physiopathogenic characteristics. PMID- 17128821 TI - [Sixty-five year old woman with sudden dysphonia, fever and productive cough]. PMID- 17128822 TI - [Giant gravidarum granuloma of the scalp]. AB - Lobular capillary hemangioma (pyogenic granuloma) has been reported more frequently in the skin (88%) than in the mucous membranes (12%). Granuloma gravidarum is a pyogenic granuloma that develops during pregnancy, often in the oral mucosa. Estrogens apparently increase the inflammatory response of mucous membranes. In this article, we describe the first case of a granuloma gravidarum of the scalp. PMID- 17128823 TI - [Antifungal and phagocytic immunostimulation treatment of chronic disseminated dermatophytosis]. AB - A 39 years old man with a long-standing disseminated dermatophytosis even with several antifungal treatments is presented. From lesions, Trichophyton rubrum var. typical downy, T. tonsurans and Candida albicans were isolated and showed sensivity to azolic compounds in vitro. The phagocytic activity in vitro compared with normal control was depressed. Treatment with itraconazole and immunomodulation using a bacterial antigen was indicated. During the last two years the patient has been clinical and mycologically healthy, and his phagocytosis activity has become normal. In patients with chronic and relapsing dermatophytosis, the immune response evaluation is recommended, and immunomodulation could be useful as a rational measure in patients with a particular immunodeficiency. PMID- 17128824 TI - [Traumatic dissection of extracranial internal carotid artery with middle cerebral artery stroke: imaging diagnosis]. AB - Cervical carotid artery dissections (CCAD) are common in young adults with a mean age of 44 years and may account for as many as 20% of strokes in patients younger than 30 years. Trauma and primary diseases of the arterial wall such as fibromuscular dysplasia are the main predisposing factors. Some CCAD cases are diagnosed solely on clinical history and physical examination, and even imaging tools such as helical/multi-slice computed tomography (CT) and magnetic resonance imaging (MRI) sometimes are not sufficient to reach a diagnosis. We describe the case of an 18-year-old male who presented to our emergency department due to loss of consciousness 18 hours after a car accident. Previously he had been in no acute distress, with fluent speech, and able to follow 3-step commands. Helical CT showed a hypodense lesion in the left-middle cerebral artery territory, as well as hyperdensity of the M1 segment of the middle cerebral artery. Cerebral angiography depicted the left carotid artery dissection in the C1 segment. Physicians should consider this entity in "asymptomatic" patients during their first hours after head injury, among patients who later develop focal neurological symptoms and clinical deficits. Clinical suspicion followed by radiological findings allows early neurovascular treatment, trying to save viable brain tissue in the first hours post injury. PMID- 17128825 TI - [Tagliacozzi: not just a plastic surgeon]. AB - Gaspare Tagliacozzi is known because of his great contributions to plastic surgery. He is considered a pioneer in the field, which has had more influence in his career than his other transcendental activity as a surgery and anatomy teacher in Bologna for almost 30 years. The aim of the present manuscript is to analyze his teaching activities which were equally important for us, and to recreate, with pictorial means, three unpublished historic moments in the life of this great man of science. Gaspare Tagliacozzi was born in Bologna in 1545 and is considered the father of plastic surgery. He obtained a degree in medicine and philosophy at the University. He was named surgery professor in 1576, and worked as such until his death in 1599. His De Curtorum Chirurgia per Insitionem treatise was published in 1589 and was considered the first exclusive treatise on plastic surgery. Bologna built a permanent operating theater (amphitheater) within the Archiginnasio in 1595. Because of his success, he increased his earnings; his first university teaching salary was 100 lire, but his earning increased to 1,140 lire later. Tagliacozzi's contributions, together with that of others such as Vesalius, Aldrovandi, Fallopian and Eustachian advanced the field and knowledge of anatomy. PMID- 17128826 TI - [Human defensins: prophylaxis and therapy against HIV?]. AB - Human defensins are endogenous antimicrobial peptides with prophylactic and therapeutic potential against HIV. The ability of defensins to bind the HIV envelope could be exploited to design topic agents that block viral entry into exposed mucosa. Additionally, their capacity to inhibit viral replication, complement system activation, dendritic and memory T cells chemoattraction, together with peptide engineering could bring about new and better antiretroviral drugs. Clinical trials could be demonstrated the efficacy of defensins against HIV in clinical practice. PMID- 17128827 TI - [Polymorphism analysis of G199A, Ncol in ANK1 and Memphis I in SLC4A1 genes in Mexican healthy individuals and subjects affected with hereditary spherocytosis]. AB - BACKGROUND: In Mexico, Hereditary Spherocytosis (HS) is the main cause of hereditary hemolytic anemia, due to mutations of one or more genes involved in the erythrocyte membrane, making it difficult to identify the primary gene. OBJECTIVE: With the purpose of estimating the use of the polymorphisms G199A and NcoI of ANK1 gene, and Memphis I of SLC4A1 gene, as genetic markers to screen this disease, we searched the allelic and genotypic frequencies in 45 DNA samples of HS patients and 28 from healthy individuals. RESULTS: Allelic and genotypic frequencies were similar in both studied groups for the G199A and Memphis I polymorphisms, with low frequency of heterozygosis showing its limited use as a genetic marker. The allelic and genotypic frequencies of the NcoI polymorphism were also similar in both groups, however a higher heterozygote frequency was observed (0.49 and 0.43 in patients and healthy individuals), a feature that may turn it into a useful genetic marker. CONCLUSIONS: Since there are other genes implicated in the molecular pathology of the HS, we consider it necessary to continue analyzing other polymorphisms of the genes involved in Hereditary Spherocytosis among the Mexican population. PMID- 17128828 TI - Medical treatment of ectopic pregnancy: a ten-year review of 106 cases at Maharaj Nakorn Chiang Mai Hospital. AB - OBJECTIVE: To evaluate the effectiveness of single-dose intramuscular methotrexate in the treatment of ectopic pregnancy at Maharaj Nakorn Chiang Mai Hospital MATERIAL AND METHOD: A retrospective review of the patient records was performed on the patients with diagnosis of ectopic pregnancy and treatment with single-dose methotrexate according to the protocol of Stovall et al between 1996 and 2005. The successful treatment was defined as no need for surgical intervention. RESULTS: As many as 96 out of 106 (90.6%) were successfully treated with methotrexate, though four required a second dose. Pretreatment /f-human chorionic gonadotropin (beta-hCG) levels were significantly lower in women who responded to single-dose therapy than either in those who required two doses or who had a failure of medical management (p < 0. 01). The mean pretreatment level of beta-hCG was 873 mlU/ml. The median time to resolution of beta-hCG was 21 days. In addition, all cases with failed medical treatment had adnexal masses larger than 3.5 centimeters in diameter CONCLUSION: In the present series, treatment of ectopic pregnancy with single-dose methotrexate was as high as 90% successful. Women with a high pretreatment beta-hCG level and large sonographic adnexal masses had a greater probability of requiring either surgical intervention or multiple doses of methotrexate. PMID- 17128829 TI - Pregnancy outcomes in placental abruption. AB - OBJECTIVES: To determine the outcomes of pregnancies with placental abruption and to investigate the relationship between clinical maternal characteristics and poor perinatal outcomes. MATERIAL AND METHOD: A retrospective descriptive study was conducted to evaluate 103 cases of placental abruption delivered at King Chulalongkorn Memorial Hospital from 1995 to 2004. RESULTS: There were 111,375 singleton deliveries with 103 cases (0.92 in 1,000) complicated by placental abruption during the study period. Placental abruption attributed to maternal complications including hemorrhagic shock (19.4%), Couvelaire uterus (16.5%) and DIC (5.8%). The perinatal outcomes included low birth weight (65.0%), preterm (56.3%), severe birth asphyxia (16.5%) and perinatal death (16.5%). Placental abruption with pregnancy induced hypertension (PIH), DIC and blood transfusion had a significantly higher incidence of perinatal mortality than the remainder (odds ratio [OR] 4.16, 95% confidence interval [CI] 1.41-12.24; OR 12.92, 95%CI 2.15-77.80 and OR 3.93, 95%CI 1.27-12.19, respectively). Placental abruption with Couvelaire uterus had a significantly higher incidence of severe birth asphyxia than the remainder (OR 3.72, 95%CI 1.14-2.09). CONCLUSION: Placental abruption had a profound impact on both maternal and perinatal complications including DIC, Couvelaire uterus, severe birth asphyxia and perinatal death. The relationship between PIH, DIC, blood transfusion and Couvelaire uterus with poor perinatal outcomes were found Therefore, placental abruption with these clinical characteristics should be closely monitored and prompt delivery should be carried out at tertiary care centers with adequate maternal-neonatal intensive care facilities. PMID- 17128830 TI - Relationship of body composition and circulatory adiponectin to bone mineral density in young premenopausal women. AB - OBJECTIVE: Adiponectin is a recently discovered hormone secreted by adipocytes. Adiponectin plays an important role in the regulation of insulin sensitivity as well as the propensity to inflammation and atherosclerosis. In the present study, the authors explore the relationship between adiponectin and bone mass in premenopausal women. The relationship of fat mass compared to lean body mass to bone mass was also investigated MATERIAL AND METHOD: Two hundred premenopausal women aged between 20 and 40 years were studied. Bone mineral density (BMD) was measured at L2-4 and femoral neck by dual-energy X-ray absorptiometry (DEXA). Serum adiponectin concentrations were measured by radioimmunoassay. RESULTS: At the lumbar spines, factors associated with BMD were age (p < 0.01) and lean body mass (p < 0.001). No independent association with fat mass was demonstrated Likewise, at the femoral neck, only lean body mass was related to BMD (p < 0. 01). In terms of the relation of serum adiponectin to BMD, no association of serum adiponectin to BMD at the lumbar spines or femoral neck was found CONCLUSION: Altogether, the present findings do not suggest the independent role of adiponectin in the accrual of bone mass in females, although such a role still cannot be excluded in men or postmenopausal women. PMID- 17128831 TI - Estimating glomerular filtration rate in Asian patients with chronic kidney diseases from bioelectrical impedance analysis. AB - BACKGROUND: Estimation of glomerular filtration rate (GFR) is usually determined from 24-hour urine collection, but it is time-consuming, and difficult in clinical practice. The authors attempted to select an accurate and safe, but more convenient test to obtain an estimated GFR. OBJECTIVE: To compare estimation of GFR by Bioelectrical impedance analysis (BIA) with GFR calculated by 24-hour urine averaged creatinine clearance and urea clearance (Ccr-Cu-GFR). MATERIAL AND METHOD: The authors examined 79 non-diabetic chronic kidney disease (CKD) patients that had estimated GFR between 15 and 89 ml/min/1. 73 m(2). Subjects were categorized into three subgroups according to K/DOQI-CKD classification: GFR of 60-89 m/min/1. 73m(2) (stage 2, 5 subjects), 30-59 ml/min/ 1.73m(2) (stage 3, 31 subjects), and 15-29 ml/min/1.73m(2) (stage 4, 43 subjects). RESULTS: The mean value of Ccr-Cu-GFR was 33.79+/-14.78 ml/min/1. 73 m(2) and GFR by BIA (BIA-GFR), 34.63 +/- 14.86 ml/min/1. 73 m(2) with no overall statistical differences (p = 0.838). In stage 3 CKD patients, the mean BIA-GFR and Ccr-Cu-GFR were similar (38.84+/-12.47 vs 41.16+/-9.17, p = 0. 399) while in stage 2 CKD, BIA-GFR tended to underestimate (63.50+/- 19.35 vs 70.94+/-7.82, p = 0.407) and in stage 4 CKD, BIA-GFR significantly overestimated Ccr-Cu-GFR (27.31+/-9.11 vs 23.76+/-5.68, p = 0.040). CONCLUSION: The findings suggest that BIA-GFR in non-diabetic CKD patients closely resembled with Ccr-Cu-GFR especially in stage 3 CKD patients. BIA-GFR may be considered as a more convenient test for an assessment of GFR in non-diabetic CKD patients. PMID- 17128832 TI - Association between waist circumference and percentage body fat among rural Thais. AB - OBJECTIVE: Determine the association between waist circumference (WC) and percentage body fat (%BF) and to develop cut-off values and evaluate the accuracy of WC in the definition of obesity in rural Thai population. MATERIAL AND METHOD: A cross-sectional, epidemiologic study in 181 men and 255 women aged 50+/-16 yr (mean+/-SD; range: 20-84 yr) sampled by stratified clustering sampling method, was designed. Percentage body fat was measured by dual energy X-ray absorptiometry (GE Lunar Corp, Madison, WI). The "golden standard"for defining obesity was%BF > or = 25 in men and%BF > or = 35 in women. Waist circumference in centimeter was measured. RESULTS: In this study, the %BF-based prevalence of obesity in men and women was 8.3% and 44%, respectively. However using the WC cut off (WHO) of 102 cm in men and 88 cm in women, only 1.7% of men and 24% of women were classified as obese. WC was a significant predictor of %BF, such that in men, a WC of 93 cm would predict a %BF of25%, and in women a WC of 84 cm would correspond to a %BF of 35%. The area under the receiver operating characteristic curve was 0. 87 and 0. 88 in men and women, respectively. In conclusion, waist circumference is a reasonably useful indicator of obesity. CONCLUSION: The cut off values of WC for diagnosing obesity should be lower in Thailand than in Western countries. PMID- 17128833 TI - Endoscopic extraperitoneal radical prostatectomy (EERPE): a new approach for treatment of localized prostate cancer. AB - Laparoscopic radical prostatectomy is usually performed by transperitoneal approach. Patients may encounter; intraperitoneal organs injury, and prolonged ileus during recovery period. The authors firstly performed endoscopic extraperitoneal radical prostatectomy (EERPE) in Thailand, which is mimicking open radical prostatectomy, the gold standard for treatment of localized prostate cancer. OBJECTIVE: Assess and evaluate the feasibility and early outcomes of the authors' experience in endoscopic extraperitoneal laparoscopic radical prostatectomy (EERPE). MATERIAL AND METHOD: From December 2005 to May 2006, 27 cases of EERPE were performed at the authors' institute for clinically localized prostate cancer by one surgeon (group I). Operative data was compared to those 55 patients who underwent open radical prostatectomy from February 2001 to August 2005 for early prostate cancer by the same surgeon (group II). Early postoperative results, clinical outcomes and complication were analyzed between the two groups using Chi-Square, student unpaired t-test and Mann-Whitney U tests. RESULTS: Patients' age and clinical staging were not different between the two groups. Mean operative time was longer in the EERPE group (268 minutes vs 157 minutes; p < 0.01). Median blood loss was 500 mls and 1000 mls in the EERPE and open groups, respectively (p < 0.001). The likelihood of transfusion rate in the open group was higher than the EERPE group, with odd ratio of 8.75 (95%CI = 2.09 39.86), p = 0.001. Hospitalization time and pathological stage were not different between the two groups. In the EERPE group, there were two rectal complications, including rectal injury and rectal necrosis, which were treated laparoscopically and conservatively without long-term problems. CONCLUSION: The authors' early experience has shown that EERPE is feasible. Although operative time was longer; the patients may gain benefit of minimally invasive surgery and decreased operative blood loss. In EERPE group, oncological outcomes are equal to open surgery, however more cases and long-term follow up are required to evaluate the efficacy of such an approach. PMID- 17128834 TI - Ramathibodi external ventricular drainage collecting device. AB - The authors report the use of Ramathibodi External ventricular drainage (EVD) collecting device. It is made of previously-used medical items. The device costs 100 baht, compared to over 3, 000 baht for those commercially available in Thailand. Over two hundred RAMA EVD collecting devices have been used with neurosurgical patients since 2002. Accurate cerebrospinal fluid (CSF) output measurement can be made with this device. Intracranial pressure monitoring is also possible. Because it is inexpensive and can be easily assembled, the authors recommend the RAMA EVD collecting device uses to other hospitals in Thailand. PMID- 17128835 TI - Atherosclerosis obliterans of the lower extremities in Thai patients. AB - BACKGROUND: Major limbs loss and high mortality rate were observed in the management of Thai patients with atherosclerosis obliterans (ASO) of the lower extremities. These were the results of delayed diagnosis and treatment together with the associated morbidities. There is a lack of information of this disease resulting in a lack of knowledge and awareness of this problem among general practitioners in Thailand. OBJECTIVES: The purposes of this study were (1) to identify the prevalence of this disease in a tertiary care hospital, (2) to enumerate the risk factors and comorbidities, (3) to identify clinical characteristics of the disease and (4) to evaluate the outcomes of treatment. MATERIAL AND METHOD: A prospective study ofpatients with ASO of the lower extremities was carried out between January 2000 and December 2004. Patients having clinical manifestations of chronic and acute arterial occlusion with the absence of ankle pulse were included in the present study. Evidence of atheromatous plaque by angiography, operative finding, and histopathology of arterial wall from amputated specimens were used to confirm the diagnosis. The selection of surgical treatments for this disease such as revascularization, major amputation, minor amputation and debridement depended on the severity of limb ischaemia, the status of distal artery and the patients' general condition. The risk factors, comorbidities, clinical manifestations, site of arterial occlusion, severity of ischaemia, types of surgical treatment and outcomes of management were analyzed. RESULTS: Four hundred and fourteen consecutive patients with ASO were diagnosed in the present study with a prevalence of 1.02:1,000. Femoro-popliteal arterial segment was the most common site (221 cases, 53.4%) of the affected arteries. Diabetes mellitus (253 cases, 61. 1%) was the most common risk factor of ASO followed by hypertension (217 cases, 52.4%), smoking (195 cases, 47.1%) and hyperlipidemia (172 cases, 41.5%). Ischaemic heart disease (108 cases, 26.1%) was the most common comorbidity of ASO followed by major stroke (56 cases, 13.5%) and chronic renal failure (20 cases, 4.8%). Patients with ASO presented mostly as chronic manifestations (385 cases, 93%) or with limb threatening condition (326 cases, 78.7%). The clinical manifestations were ischaemic ulcer and/or digital gangrene (251cases, 60.6%), rest pain (182 cases, 44.0%) incapacitating claudication (62 cases, 15.0%) and acute ischaemic pain (29 cases, 7.0%). One hundred and thirty eight (33.3%) patients had significant lower limb infection at the time of admission. One hundred and seventy one (41.3%) patients underwent revascularization procedures as the major primary treatments to salvage the limbs. The success rate of limb salvage after revascularization was 76.6% (13 1/171). Major amputation after revascularization was 16. 9%(29/ 171). Perioperative mortality rate of revascularization procedure was 8.2 %(14/171). Major amputation was required as the primary treatment due to infective (18.4%, 76/414) and ischaemic process (6.5%, 27/414). The mortality rate of primary major amputation for infection and ischaemia were 19.7%(15/76) and 25.9%(7/2 7) respectively. The total mortality rate in the present study was 11.3% (47/414). The common causes of death were sepsis and ischaemic heart disease. CONCLUSION: ASO of the lower extremities is one of the major problems for national health care causing major limb loss and death. Arterial bypass surgery was the most effective treatment for limb salvage. Management of this disease at the terminal stage causes high morbidity and mortality. Hence, early detection of this disease and correction of the risk factors should be the most effective strategy to improve the overall outcome of the management of this complicated problem. PMID- 17128836 TI - Economic problem of referred trauma cases in Siriraj Hospital. AB - To improve the service for referred trauma patients, the authors have to know the types of injury, the cost, and the reimbursement for each individual category. Therefore, the authors studied the characteristics of accident of patients transferred for treatment at Siriraj Hospital, including types of trauma (wounds), cost of treatment (excluding doctor's fees) and charges for treatments for each group. There were 678 trauma cases referred to the Division of Trauma Surgery, Department of Surgery, Faculty of Medicine, Siriraj Hospital from 1st September 2002 to 31st August 2003 and of these, 571 cases were admitted From the admitted cases, the most common injury was the musculoskeletal system (38.6%), followed by nervous system injury (15.1%), eye injury (8.1%), toxic substance injury (6.3%), burns (5.8%), and others. The total cost of treatment was 36,249,150 baht. The reimbursement was 30,135,709 baht. There were four categories where the reimbursed amount was below the actual cost. They were burns, followed by nervous system injury, eye injury and skin or subcutaneous tissue injury. To deal with the referred cases in trauma center level I, the center should prepare to manage the common trauma groups such as musculoskeletal system injury, nervous system injury and eye injury. Burns, nervous system injury, eye injury and skin or subcutaneous tissue injuries are the major groups that cost more than the reimbursement amount. The reimbursement of these groups should be reconsidered in the future to solve the problem. PMID- 17128837 TI - Neurovascular anatomy of the deep inferior epigastric perforator flap for breast reconstruction. AB - OBJECTIVE: To find out the most suitable perforator and cutaneous nerve for strategic design of the deep inferior epigastric perforator (DIEP) flap. MATERIAL AND METHOD: The characteristics of the pedicles, perforators, intercostal nerves and the relationship between nerves and vessels in DIEP flaps were studied in 31 formalin-preserved cadavers. RESULTS: Four hundred and five perforator vessels were divided into three vertical rows. These perforators were mostly contained in the medial row (45.4%), the average size of the perforators in the lateral row was the largest (1.0+/- 0.3 mm). The largest perforators (1.4+/- 0.3 mm) were mostly located within 1 cm horizontally from the umbilicus. Lateral row perforators, usually rectilinear course (82.7%), traveled with nerves from the beginning. Whereas, the perforators in the medial row usually coursed obliquely (86.4%) and were not related to nerves initially, they joined before piercing the rectus sheath. CONCLUSION: The present findings indicate that it would be more beneficial to use the lateral row perforators. PMID- 17128838 TI - Prevalence and clinical features of mycoplasma pneumoniae in Thai children. AB - OBJECTIVE: To determine the prevalence and clinical features of mycoplasma pneumoniae in Thai children with community acquired pneumonia (CAP). MATERIAL AND METHOD: Diagnosis of current infection was based on > or = 4 fold rise in antibody sera or persistently high antibody titers together with the presence of mycoplasma DNA in respiratory secretion. The clinical features were compared between children who tested positive for M pneumoniae, and those whose results were negative. RESULTS: Current infection due to M. pneumoniae was diagnosed in 36 (15%) of 245 children with paired sera. The sensitivity and specificity of polymerase chain reaction (PCR) in diagnosing current infection in the present study were 78% and 98% respectively. The mean age of children with mycoplasma pneumoniae was higher than CAP with unspecified etiology. The presenting manifestations and initial laboratory finding were insufficient to predict mycoplasma pneumoniae precisely, the presence of chest pain and lobar consolidation on chest X-ray, however, were significant findings in children with mycoplasma pneumoniae. CONCLUSION: The present study confirms that M. pneumoniae plays a significant role in CAP in children of all ages. Children with this infection should be identified in order to administer the appropriate antibiotic treatment. PMID- 17128839 TI - Low-dose intradermal and subcutaneous versus intramuscular hepatitis B vaccination in primary non-responding hemodialysis patients. AB - OBJECTIVE: Patients with end-stage renal failure are at high risk of hepatitis B virus (HB V) infection. They have impaired immune response to HBV intramuscular (i.m.) vaccine. Non-response (anti HBs titer < 100mIU/ml) hemodialysis patients (HD) with the previous three-dose i.m. vaccination were examined with booster dose vaccine by i.m. , intradermal (i.d) and subcutaneous (s.c.) routes. MATERIAL AND METHOD: Thirty-four HD patients who had been vaccinated with three-dose vaccine (40 microgram, 2 ml, Engerix B, i. m.) and had anti-HBs titer less than 100mlU/ml were selected. They were randomly divided into three groups and received a fourth dose of vaccine by i.m. (40 microgram, 2 ml), i.d (10 microgram. 0. 5 ml) and s.c. (10 microgram, 0. 5 ml). Then, serum anti-HBs titer was determined after 45 days and 6 months. RESULTS: Forty five days after completion of the re-vaccination course, anti-HBs titer was above 100 mIU/ml in 6/11, 3/11 and 4/12 of i.m. s.c. and i. d groups, respectively (p > 0.05). After six months, 4/11, 3/11 and 2/12 of patients had anti-HBs titer above l00mlU/ml (p > 0.05). CONCLUSION: With lower dose of vaccine (10 microgram) in s.c. groups, these patients had lower change in their anti-HBs titer. Therefore, it is cost effective and practical to offer other vaccination schemes. PMID- 17128840 TI - Comparison of dexpanthenol and zinc oxide ointment with ointment base in the treatment of irritant diaper dermatitis from diarrhea: a multicenter study. AB - BACKGROUND: Severity of irritant diaper dermatitis (IDD) from diarrhea varies from patient to patient depending on the nature of feces and the number of bowel movements. The purpose of the present study was to compare the effectiveness of dexpanthenol and zinc oxide ointment with ointment base in the treatment of irritant diaper dermatitis from acute diarrhea in children by measuring transepidermal water loss (TEWL). MATERIAL AND METHOD: Forty-six children with diarrhea were prospectively, block randomized, investigator-blinded to receive dexpanthenol and zinc oxide ointment on one side and ointment base on the other side. TEWL was measured before and on days 1, 3, and 7 of treatment together with the assessment of severity score. The efficacy of treatment was defined by complete clearance of the lesion. RESULTS: TEWL in the treated and control side was not different before the application of the topical medication. In the present study, the efficacy of 5% dexpanthenol and zinc oxide ointment on D3 was 39% (18from 46 patients) compared to 32% in the ointment base side. On D7, the efficacy of the treated side was 58.7% and the ointment base side was 56%. The patients who still had skin lesions were those who had prolonged diarrhea. On the treated side, the mean of TEWL was lower than the control side on DI (p = 0.18) and had significant improvement on D3 (p = 0. 002). At the end of the present study, TEWL on the treated side was less than TEWL of the control side but it did not have statistical significance (p = 0.07). There was no rash or sign of abnormality on the treated side at the end of D7. CONCLUSION: In the treatment of lDD from acute diarrhea, 5% dexpanthenol and zinc oxide ointment significantly decreased TEWL in the treated side more than the ointment base on day 3 but the severity score was not significantly different on days 1, 3 and 7. PMID- 17128841 TI - Outcomes of scleral buckling for stage 4 retinopathy of prematurity in Thai children. AB - OBJECTIVE: To evaluate the anatomical and visual results of a primary scleral buckling procedure for the treatment of stage 4A and 4B retinopathy of prematurity (ROP) in Thai patients. MATERIAL AND METHOD: The data of premature infants treated with a primary scleral buckling procedure for stage 4 ROP from December 2000 to May 2004 were retrospectively reviewed The outcomes measures were anatomical success, visual outcomes, and refractive error at the end of follow-up. RESULTS: Sixteen eyes of ten patients underwent a scleral buckling procedure and had the mean follow-up period of 17.3 months (range 3-44 months). The anatomical success was 100% (8 of 8 eyes) in stag 4A and 50% (4 of 8 eyes) in stage 4B. At the end of the follow-up, the buckle was removed in 92% (11 of 12 eyes) of retina-attached eyes and showed a mean myopic refraction of-8.68 diopters (range -4. 75 to 13.50). Favorable visual outcome was 50% (4 of 8 eyes) in stage 4A and 12.5% (1 of 8 eyes) in stage 4B. CONCLUSION: Scleral buckling appears to play a role in reducing the progression from stage 4 to stage 5 ROP The anatomical success rate was excellent but the visual results remain challenging for these cases. PMID- 17128842 TI - Stereotactic radiation therapy for optic nerve sheath meningioma; an experience at Ramathibodi Hospital. AB - OBJECTIVE: To evaluate results of stereotactic radiotherapy for the treatment of optic nerve sheath meningioma (OM) at Ramathibodi Hospital. MATERIAL AND METHOD: Twelve patients with primary OM were treated with stereotactic radiation between 1998 and 2005. Five patients underwent surgery and had no light perception before radiation. All patients except one were treated with fractionated stereotactic radiotherapy (FSRT). Mean average dose of FSRT was 55. 7 Gy; 180 cGy/fraction. One patient was treated with 15-Gy stereotactic radiosurgery. RESULTS: With a median follow-up of 34 months, there was no visual improvement in the five patients who were completely blind before radiation. Visual acuity improved in four patients and remained stable in two patients. Four of six patients had improved visual field, and five of six decreased in proptosis. Follow-up images were available in six patients, showing minimal tumor regression in five and stable in one. No serious acute side effect was observed. Vision became worse in one patient, who developed vitreous hemorrhage two years after FSRT. CONCLUSION: Stereotactic radiotherapy is an effective treatment for primary OM. It provides tumor control and visual preservation with low risk of complications. However more patients and further follow-up are needed for long-term outcomes. PMID- 17128843 TI - Direct immunofluorescence study in Thai patients with scleroderma. AB - OBJECTIVE: Previous reports of direct immunofluorescence (DIF) studies of the skin biopsies in scleroderma were either negative or positive at various percentages and patterns. The present study was designed to evaluate the positive yield and pattern of DIF in Thai patients with scleroderma and its possible clinical correlation. MATERIAL AND METHOD: Twenty-two patients with localized or systemic sclerosis, who attended the Department of Dermatology, Siriraj Hospital, from 1996 to 2002, were enrolled in the present study. Skin biopsy was performed for DIF studies. RESULTS: Nine out of 22 patients were diagnosed with systemic sclerosis (SS), eleven with morphea, and two with overlapping syndrome. Fifteen of 22 patients (68%) had positive DIF findings; seven of nine (78%) patients with SS, six of eleven (55%) patients with morphea and two of two (100%) with overlapping syndrome. The common sites of deposit in SS, morphea and overlapping syndrome were dermo-epidermal junction and epidermal nuclei. The common immunoreactant deposit in all groups was IgM. There was no significant difference in the comparison of DIF findings with duration of biopsy lesion, clinical correlation, and the positive result of serum antinuclear antibody (ANA) in the three groups of patients. CONCLUSION: Positive DIF yield in the present study was higher than previous reports from Western countries. Similar to the study reported from Western country, there was no statistical significant difference in comparison of DIF findings with the duration of lesion, clinical correlation, and the positive result ofserum ANA in our three groups of patients. However; patients with SS had a tendency to give more frequently positive ENS and DEJ deposits than those with morphea. PMID- 17128844 TI - Comparison of one week with two week regimens of amphotericin B both followed by fluconazole in the treatment of cryptococcal meningitis among AIDS patients. AB - BACKGROUND: Amphotericin B treatment in cryptococcosis requires daily hospital visits or admission. Its toxicities and hospital costs have been concerned. Short course amphotericin B regimen warrants to be evaluated. OBJECTIVE: To compare the safety and efficacy of one-week (AmB1) with two-week (AmB2) amphotericin B both followed by fluconazole. MATERIAL AND METHOD: 57 AIDS with cryptococcal meningitis were randomly assigned to either AmB1 or AmB2. Microbiological and clinical clearances were the outcomes of the study. RESULTS: The treatment success at 6 weeks was 63.3% in AmB1 and 70.4% in AmB2 (p = 0.574). Clinical assessment at week 10 and renal toxicities were not significantly different between both regimens. Mortality rate was 14% however, 75% of deaths were in AmB2. CONCLUSION: AmB1 was comparably effective and safe as the standard AmB2 regimen in the treatment of AIDS related cryptococcal meningitis. It can be an alternative regimen to lower hospital based care and improve cost effective for source limiting health care centers. PMID- 17128845 TI - Lipid peroxidation and renal injury in renal ischemic reperfusion: effect ofangiotensin inhibition. AB - OBJECTIVES: To investigate the role of angiotensin inhibition on lipid peroxidation (LPO) and renal pathology in ischemic reperfusion (IR). MATERIAL AND METHOD: Male Wistar rats were subjected to 15-, 30-, 45- or 60- minutes ofrenal ischemia (I) by left renal artery occlusion. In the 30-minute I group, reperfusion (R) for I day (13,R) was performed in additional animals that had been treated with water, angiotensin converting enzyme inhibitor (ACE]; enalapril 5 mg/kg/d), or angiotensin receptor type 1 blocker (ARB; losartan 10 mg/kg/d) one day before I and were continued for 1 day after R. Renal tissue malondialdehyde (MDA), an indicator of LPO, was examined during I and IR periods. Renal pathology was also determined. RESULTS: During ischemia, renal tissue MDA levels were increased throughout the 60-minute ischemic period and was maximum at 30 minutes of ischemia (p < 0.01). Histological changes in 30-minutes I group showed slight tubular cell congestion and mild interstitial edema. One day after reperfusion, MDA levels were still elevated (p < 0. 01) when compared with sham. Progression of renal pathology was observed after I day of reperfusion. Both ACEI and ARB could attenuate the heightened MDA levels (p < 0.01). IR-induced renal injury was markedly diminished by administration ofACEI as well as by ARB. CONCLUSION: These results indicate that inhibition of angiotensin could reduce lipid peroxidation and ameliorate renal injury during IR condition. PMID- 17128846 TI - Cross-cultural adaptation of modified Oswestry Low Back Pain Disability Questionnaire to Thai and its reliability. AB - OBJECTIVE: The present study aimed to cross-culturally adapt the modified Oswestry Low Back Pain Disability Questionnaire (ODQ) into Thai. MATERIAL AND METHOD: The process comprised of an initial forward translations from English to Thai, synthesis of the translations, back translation, and back translation approval. The approved version of Thai ODQ was then calculated for test-retest reliability. Forty patients with LBP, aged 40.1+/-10.7 years, were recruited into a test-retest reliability study. RESULTS: The test-retest reliability, calculated by intraclass correlation coefficient, was assessed on two occasions separated by a time interval of 20-30 minutes. The values of test-retest reliability of items ranged from 0.80-1.00. The value of total score was 0.98. CONCLUSION: This finding indicated good reliability of the Thai version modified ODQ. PMID- 17128847 TI - Normal internal organ weight of Thai adults correlated to body length and body weight. AB - OBJECTIVE: Examine the relationship between the internal organ weight with body weight and body length. MATERIAL AND METHOD: Analysis of data from 250 autopsies from the Ramathibodi Hospital from August 2003 to February 2005. The cases were from sudden unnatural death including accident, homicide and suicide and excluded decomposed bodies, fire related deaths and cases where medical treatment had been given. The age ranged from 15 to 88 years and there were 51 females and 199 males. Parson's correlation coefficient was used to examine the relationship between the internal organ weight with body weight and body length. RESULTS: The mean+/-standard deviation (SD) were represented by males and females respectively; Brain 1339+/-160/1165+/-184 gm, heart 311+/-66/278+/-160 gm, lung 910+/-347/675+/-255 gm, liver 1439+/-365/ 1214+/-275 gm, spleen 103+/-46/92.9+/ 48 gm, kidney 260+/-68/230+/-42 gm. CONCLUSION: The relationship between internal organ weight and body weight showed each internal organ significantly correlated with body weight in males at p-value < 0.05, whereas in females it only correlated to liver, kidney and spleen at p-value < 0.05. For the correlation between internal organ weight and body length, it showed only brain, lung, liver and kidney correlated to the body length in males at p-value < 0. 05, but not in females. PMID- 17128848 TI - Multi-drug resistant HIV-1 reverse transcriptase genotype in children treated with dual nucleoside reverse transcriptase inhibitors (NRTIs). AB - BACKGROUND: Multi-drug resistant HIV mutants have been reported after prolonged dual antiretroviral therapy. OBJECTIVE: To evaluate the prevalence and resistance pattern in HIV-infected children treated with dual NRTIs. MATERIAL AND METHOD: Records of HIV-infected children treated with dual NRTIs at Srinagarind Hospital, Khon Kaen University, Thailand, were reviewed for baseline data and their consensually-stored plasma were checked for the occurrence of HIV mutants by genotyping. RESULTS: Fifty-seven HIV-infected children were treated with dual NRTI regimens (27 males; 30 females). The median age and median CD4+ T-lymphocyte at genotypic testing were 83.5 months and 10.9%, respectively. The median duration of ARV therapy was 22 months. More than half the children (42) were on zidovudine and didanosine. A set of three or more nucleoside analog mutations (NAMs), conferring multi-dideoxynucleoside resistance, was found in 60% of the cases. CONCLUSION: High percentages of NAMs were found in HIV-infected children previously on dual ARV therapy for long periods. Genotypic testing was helpful in designing the second antiretroviral regimen. PMID- 17128849 TI - HIV/sexual and reproductive health program for HIV prevention: the youth-adult partnership with schools approach. AB - The aim of the present study was to describe the development and evaluation of a program designed to prevent HIV/AIDS. A participatory action research (PAR) approach was used in collaboration with ten schools in Chiang Mai Province, Thailand, to develop a youth- adult partnership with schools (YAPS) model. The YAPS model included curricula using participatory learning experiences, edutainment approaches, and skills building strategies for enhancing youth leaders'capacities. Results showed that the YAPS model was effective in leadership role preparation and in empowering youth leaders to undertake activities on their own, initiate creativity and share knowledge on sexuality education and HIV prevention messages with students in schools. The use of partnerships and the participatory process mobilized parents, teachers, and school administrators to play a proactive role in sexuality education and HIV prevention for early adolescents in schools, resulting in the integration of the program into the school system. PMID- 17128850 TI - Nosocomial tuberculosis risk assessment and management recommendation for an emergency department in a university hospital. AB - OBJECTIVES: To assess the nosocomial TB risk and recommend appropriate risk management procedure for the trauma ward of the emergency department at a university hospital in Bangkok. MATERIAL AND METHOD: The risk assessment procedure includes: (1) estimating the number of healthcare visits of the TB patients at the trauma ward of emergency department in 2004; (2) determining the TB incidence rate among the healthcare workers (HCWs) of the trauma ward during 2004-2005; and (3) surveying the indoor environment (particularly concerning the ventilation condition) of the targeting ward. Appropriate risk management procedures were then recommended based on the guidelines provided by The World Health Organization (WHO) and The Centers for Disease Control and Prevention (CDC) of the United States. RESULTS: The estimated number of healthcare visits of the TB patient at the trauma ward in 2004 was 20. During 2004-2005, there were 4 out of 57 HCWs at the trauma ward who developed tuberculosis; among these were two registered nurses and two nursing auxiliaries. The TB incidence rates among these HCWs were 3,509 per 100,000 person-year overall (or 2,632 per 100, 000 person-year for confirmed TB cases only) and 3,333 (or 1,667 for confirmed TB cases only) and 4,761 per 100, 000 person-year for registered nurse and nursing auxiliary. The ventilation in the trauma inpatient ward had 1.43 air changes per hour, which was below the recommended standard value. Turbulence pattern of airflow, which promotes airborne TB spreading in this area, was also detected. CONCLUSION: The risk assessment results showed that the trauma ward of the emergency department at this university hospital has a high risk for nosocomial tuberculosis. Appropriate risk management procedures, which are in accord with the standard guidelines, should be urgently implemented. PMID- 17128851 TI - The development of a computer based learning (CBL) program in diabetes management. AB - OBJECTIVE: To develop a computer based learning (CBL) program in diabetes management for health care providers and academic staff MATERIAL AND METHOD: A CBL program was developed using "Authorware Professional ver. 6. 0" software. Content validation, computer background survey and investigation of the usability of the program, was conducted as part of the production of this program. The involved participants were university staff hospital care providers (e.g., doctors, nurses and pharmacists) and nursing & pharmacy students. RESULTS: Overall, the results were positive. Some limitations regarding computer background were revealed Few of the participants were familiar with self-learning materials. The usability of the CBL program was generally encouraging however some comments were made regarding program function, such as the duration of the program, and minor problems with the audiovisual effects. All comments were noted and addressed for future implementation. CONCLUSION: The CBL program was found to be a user-friendly, interactive multimedia program for diabetes management. PMID- 17128852 TI - Hand-assisted laparoscopic partial nephrectomy for renal cell carcinoma: the first case report in Thailand: case report. AB - The authors report their first experience on a hand-assisted laparoscopic partial nephrectomy surgical technique and the result after it was performed successfully in a 45-year-old woman who presented with an asymptomatic small renal mass. This is the first successful case report of this operation in Thailand. PMID- 17128853 TI - Primary umbilical endometriosis. AB - A 45-year old nulliparous woman presented with umbilical pigmented lesion with cyclical bleeding. The lesion was excised and pathological diagnosis was umbilical endometriosis. The authors reviewed the current literature and discussed the different diagnosis and management of umbilical endometriosis. PMID- 17128854 TI - Cystic fibrosis in three northeast Thai infants is CF really a rare disease in the Thai population? AB - The authors report on three infants with cystic fibrosis (CF), with different genotypes, presenting with different clinical manifestations, but having similar abnormal serum electrolytes (i.e. hyponatremia, severe hypochloremia and metabolic alkalosis). Despite the diagnostic investigations, the child who presents with severe electrolyte imbalance especially persistent hypochloremia and a family history of early infant death with respiratory or gastrointestinal problems should point to a diagnosis of CF Early identification and treatment remain critical to effective management. The diagnostic tool used, especially the sweat test, is needed for diagnostic investigations in Thailand. PMID- 17128855 TI - Real-time three dimensional sonographic features of an early third trimester fetus with achondrogenesis. AB - Generalized shortening of fetal long bones detected from prenatal sonographic examination usually raise a tentative diagnosis of skeletal dysplasia. Information obtained from grey-scale scan is frequently not sufficient to provide a definite diagnosis, and the images are not readily comprehensible for the parents-to-be. Lately, three-dimensional sonography has become increasing available in obstetric practice. The authors report here a rare case of fetal achondrogenesis, which is a lethal form of skeletal dysplasia, in a 30-week-old fetus using real-time three-dimensional ultrasound. The prenatal findings of fetal achondrogenesis from this technique were thoroughly described, along with postnatal radiography and autopsy results. Sonographic features from this imaging technique allow for an accurate diagnosis and better understanding of the parents. This facilitates the genetic counseling process, as well as the parental options for further care. PMID- 17128856 TI - Corneal injury by bee sting with retained stinger--a case report. AB - OBJECTIVE: To report a rare case of corneal injury by bee sting with its complication and management. MATERIAL AND METHOD: A 3-year-old boy, who was attacked by a swarm of bees, was referred for the right eye's corneal ulcer evaluation. RESULTS: Two stingers were found and completely removed with jeweler forceps. Corneal epithelial defect, corneal edema, secondary bacterial keratitis, heterochromia iridis, and internal ophthalmoplegia were identified The corneal edema markedly improved 3 days after removing the retained bee stingers and treatment by topical antibiotics and steroids. The patient was evaluated after 1 week and 1 month and was found with permanent central corneal scar particularly at the area corresponding to the retained stinger The patient had subsequently localized traumatic anterior subcapsular cataract corresponding to where the stinger had penetrated the lens. CONCLUSION: Corneal bee sting injury is an uncommon ocular trauma, but can result in severe sight threatening complication. Even though the response of corneal edema is well inclined to topical steroid, awareness in adjusting the clinical treatment for the particular case needs a scrutinized investigation of the infection. PMID- 17128857 TI - Hepatic fascioliasis due to Fasciola hepatica: a two-case report. AB - Two cases of hepatic fascioliasis due to Fasciola hepatica were retrieved from our surgical-pathology file since the hospital's foundation in 1969 up to 2005. The diagnosis of hepatic fascioliasis was based on detection of one live fluke in a large cystic lesion in the lobectomized liver specimen in one case and of deposited eggs in the large liver specimen obtained from open biopsy in the other Hepatic fascioliasis is rather rare and almost worldwide in distribution including Thailand. The diagnosis should be considered in the patient from endemic areas consisting of the northern, northeastern and upper-central regions of the country, with a history of ingesting fresh water plants or drinking untreated water and having fever right-upper-quadrant pain or intrahepatic cystic lesion(s) together with absolute peripheral blood eosinophilia. PMID- 17128858 TI - Specific language impairment: effect on later language development: a literature review. AB - BACKGROUND: Children with specific language impairment (SLI) are at considerable risk for later language development. However there has not been a good literature review to provide understanding and to get information relative to parental counseling and decision making for further management. OBJECTIVE: To summarize the literature review on SLI and effect on later language development. MATERIAL AND METHOD: Longitudinal studies were systematically explored to seek evidence- based information to confirm the lasting effect of SLI in later language development. RESULTS: Natural history data indicated that approximately 50% of children with specific expressive language delay spontaneously remitted or outgrew it at age 5-8. However, long-term follow-up studies showed their language problems emerged later at age 15. The rest of the children's (50%), language impairment persisted and had a high risk for SLI later. CONCLUSION: The results suggest that active intervention should be performed as soon as possible for children with persisting SLI. PMID- 17128859 TI - Regulation of stem cell fate in hematopoietic development. AB - Stem cells are broadly classified into two categories; embryonic stem cell and somatic stem cell. Both types of stem cells can be differentiated into many cell types of the body with a different capability. Blood cells are examples of the cells that can be derived from both embryonic and somatic stem cells, both in vivo and in vitro. In adults, blood cells are mainly derived from hematopoietic stem cells (HSC) in the bone marrow. HSC growth and differentiation is tightly regulated which require both intrinsic and extrinsic signals. In this regard, transcription factors are critical for the development of HSC and specific cell lineages, in part, by regulating the expression of hematopoietic growth factor (HGF) receptors, other transcription factors and lineage specific genes transcription. This review will focus on the role of Id transcription factors in stem cell fate regulation. PMID- 17128860 TI - Health care personnel ethics. PMID- 17128863 TI - Texas case spotlights end-of-life quandary. PMID- 17128865 TI - In from the cold. PMID- 17128866 TI - Lung cancer. The battle continues. PMID- 17128867 TI - What's wrong with this patient? Primary systemic amyloidosis. PMID- 17128868 TI - Solving puzzles. Interview by Jennifer Samuels. PMID- 17128869 TI - Who has a forklift? PMID- 17128870 TI - [Simultaneous determination of pesticide residues in crops by liquid chromatography with tandem mass spectrometry]. AB - A simultaneous method using liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) was developed for the determination of pesticide residues in crops. Mass spectral acquisition was performed in the positive mode by applying multiple reaction monitoring. In LC separation, an Atlantis dC18 Column was used with acetic acid-ammonium acetate acetonitrile as the mobile phase. Pesticide residues in crops were extracted with acetone, and cleaned up by liquid-liquid separation with saturated salt solution and hexane, followed by an ENVI-Carb cartridge. The quantification limits of compounds in crops were below 5 ng/g. Eighty compounds were obtained with recoveries ranging from 60 to 130% at the level of 50 ng/g with RSD (%) of less than 15%. Fifty crop samples were analyzed by the developed method. Seven pesticide residues were detected in nine crops. PMID- 17128871 TI - [Screening method for the determination of 199 pesticides in agricultural products by gas chromatography/ion trap mass spectrometry (GC/MS/MS)]. AB - A screening method is described for determining 200 pesticides, except dimethipin, divided into four groups by means of gas chromatography/tandem mass spectrometry (GC/MS/MS) using an ion trap mass spectrometer equipped with automated gain control (AGC). The quantitation limit for 194 pesticides was 0.01 mg/kg on a crop basis, except for allidochlor, dimethoate, hexythiazox, methamidophos and triadimenol. The calibration curve of each pesticide was linear in the range of 0.04-5.0 microg/mL. One hundred and ninety-nine pesticides were added to matrix of potato, spinach, cabbage, apple, orange, soybean and unpolished rice at twice the limits of quantitation. The recoveries of 194 pesticides from all crops were satisfactory (50-150%) for screening purposes. Although some pesticides in apple and orange were not determined by selected ion monitoring (SIM) analysis at the limits of quantitation, all of them were identified by ion-trap GC/MS/MS at the same concentration. Thus, the ion trap GC/MS/MS technique is useful for the screening of residual pesticides present at low levels in agricultural products. PMID- 17128872 TI - [Detection of fish DNA in ruminant feed by PCR amplification]. AB - The Japanese Government has prohibited the use of seafood protein, as well as mammalian protein, in ruminant feed. There is an official method to detect meat and bone meal, but no method is yet available to detect fishmeal in ruminant feed. We tried to develop a suitable method to detect fishmeal in ruminant feed, similar to the official method "PCR detection of animal-derived DNA in feed". Our previously reported primers (fishcon5 and fishcon3-1) showed low sensitivity, so we designed new primers based on a DNA sequence from yellowfin tuna mitchondrial DNA. Among the primers, FM5 and FM3 specifically detected fish DNA (sardine, yellowfin tuna, skipjack tuna, chub mackerel, Pacific saury, salmon, rainbow trout, Japanese anchovy, codfish and Japanese horse mackerel) from fish meat, and did not amplify DNA from animals and plants. The sensitivity for detection of the presence of fishmeal in ruminant feed was 0.01-0.001%. PMID- 17128873 TI - [Applicability of GC, GC/MS and liquid chromatography with tandem mass spectrometry to screening for 140 pesticides in agricultural products]. AB - The applicability of GC, GC/MS and liquid chromatography with tandem mass spectrometry (LC/MS/MS) to screening for 140 pesticides in agricultural products was examined. Validation of multi-residue screening methods for the determination of 88 pesticides in 12 crops (asparagus, cauliflower, burdock, carrot, broccoli, spinach, matsutake mushroom, orange, soybean, sesame, millet and tea) was done by GC and GC/MS. Of the 88 pesticides, 63 were obtained with recoveries in the range from 50 to 150% at the 0.1 microg/g level in the 12 crops. Applicability of the official methods in Japan to 74 pesticides, including 22 pesticides with low recovery (< 50%) by GC or GC/MS analysis, was also examined by LC/MS/MS. LC/MS/MS acquisition parameters were established for 67 pesticides in positive and negative electrospray ionization (ESI) modes. Of 67 pesticides validated in 7 crops using LC/MS/MS at the 0.1 microg/g level, 44 showed recoveries in the range from 50 to 150%. The occurrence of matrix interference in LC/MS/MS can lead to false-positive detection of MCPA in spinach, cabbage and orange and false negative detection of four pesticides in orange, spinach, apple and unpolished rice. Good linearity was observed in the studied ranges by GC, GC/MS (r > 0.990) and LC/MS/MS (r > 0.995). Of the total of 140 pesticides validated by GC, GC/MS and LC/MS/MS, 107 were newly recognized as suitable subjects for screening. PMID- 17128874 TI - Standard infrared absorption spectrum of betaine and optimal conditions for its measurement. AB - The infrared absorption (IR) spectrum is often used as a standard reference in identification tests of food additives in Japan. In the case of betaine, many different IR spectra have been reported and, therefore, it is necessary to establish an IR spectrum that is reproducible and reliable enough to be used as a standard for identification. In the present study, suitable conditions to obtain a standard IR spectrum were examined from various viewpoints, including pretreatment, selection of method, and measuring technique. The KBr disk method, which has generally been used to identify betaine, was found to be humidity dependent, and there was also an interaction between betaine and KBr. A reproducible IR spectrum suitable as a standard could be obtained by drying betaine at 105 degrees C for 3 hours over phosphorus pentoxide, and then measuring the IR spectrum by the liquid paraffin (Nujol) paste method. PMID- 17128875 TI - [Bursting risk of capped PET bottles with leftover contents caused by yeast growth]. AB - Accidents caused by the bursting of capped PET bottles with leftover contents have increased in Japan. Therefore, the bursting risk of capped PET bottles with leftover contents was investigated. The ratios of distension, after 10 days at 25 degrees C, of capped PET bottles with leftover orange juice were approx. 70% in the case of drinking while eating Kimuchi, and approx. 6% in the case of drinking but not eating. Among 58 distended bottles, none burst spontaneously, but 4 bottles (6.9%) burst upon physical shock (dropping onto a concrete floor from 1.5 m height). The greatest distensions of capped PET bottles were observed with leftover orange juice, apple juice and soured milk after 10 days at 25 degrees C. No distensions were observed in bottles containing leftover sports drink and green tea under the same conditions. Identified organisms from distended bottles were almost all Candida spp. When those Candida spp. strains were inoculated individually into PET bottles half full of orange, apple or sour milk, all bottles were distended after 7 days at 25 degrees C. From the above results, it appears that there is a risk that capped PET bottles with leftover contents, especially sour fruit juice or soured milk, may burst owing to high gas pressure caused by yeast growth during storage at room temperature for over 1 week. PMID- 17128876 TI - [Application of dual column GC for pesticide residues analysis in foods]. PMID- 17128877 TI - [Study on safety information of health food]. PMID- 17128878 TI - [New trends in carcinogenic thresholds---International Symposium on "Thresholds for Carcinogens and Mutagens"]. PMID- 17128879 TI - [Study of development of analytical methods of veterinary drugs and other chemicals in food]. PMID- 17128880 TI - [Studies of food hygiene about decomposition product from pesticides detected in foods]. PMID- 17128881 TI - [Contamination levels of dioxins in foods and cleanup methods for the determination]. PMID- 17128882 TI - [Survey on the current status of delivered informed consent for renal replacement therapy among patients with end-stage renal disease]. AB - We conducted a survey on the adequacy of delivered informed consent (IC) among patients with end-stage renal disease (ESRD) regarding the information provided on renal replacement therapies (RRT): Hemodialysis (HD), peritoneal dialysis (PD), and renal transplantation (RTx). A self-assessment style of questionnaire entitled "Informed consent for the selection of dialysis therapy modality" was prepared for evaluation, and the adequacy of IC was scored by 5 grades ranging from "excellent" to "bad". The questionnaire was sent to all the JSDT registered facilities (n=3484), and 480 centers replied (13.8%). Among these, 407 centers had patients introduced onto some form of RRT modality in the last 12 months. As to the adequacy of delivered IC for each modality, "excellent and good" status was reported as follows: 80.8% in HD, 49.8% in PD, and 32.5% in RTx, respectively. The major reason for "poor and bad" IC adequacy in PD and RTx, was "not available in the facility". By analyzing the facilities stratified by the clinical experiences of each modality in the past, poorly delivered IC for PD and RTx was revealed in centers lacking experience. Delivered information about RRT to ESRD patients may be biased in Japan. The findings of this study suggested that a lack of medical experience of the modality contributes to insufficient IC. PMID- 17128883 TI - [Significance of urine dipstick test for occult blood in patients with diabetes mellitus]. AB - OBJECTIVE: Hematuria is the usual urinary finding in patients with diabetic nephropathy. In this study, prevalence and significance of microhematuria in patients with diabetes mellitus were investigated. MATERIAL AND METHODS: Five hundred and forty-two samples from outpatients, who consulted the Department of Diabetes and Metabolism, were categorized into 4 groups according to the results of the urine dipstick test. Group I :258 samples, urinary protein(u-P) 1+ and occult blood (OB) negative, group II: 95 samples: u-P 1+, OB 1+ approximately 3+, group III: 89 samples: u-P 2+ approximately 3+, OB negative, group IV 100 samples u-P 2+ approximately 3+, OB 1+ approximately 3+. RESULTS: Serum concentrations of creatinine and highly sensitive CRP were both significantly higher in group II than in group I . Those parameters were also significantly higher in group IV than in group III. CONCLUSIONS: These results indicate that a positive dipstick test for OB may occur in many patients with diabetes mellitus and suggest the progress of renal damage. PMID- 17128884 TI - [Cerebral salt wasting syndrome in a patient with viral meningoencephalitis]. AB - A 53-year-old male was admitted to our hospital for a high fever. He suffered a change in personality, memory loss and disorientation as well. The findings of cerebrospinal fluid showed monocytosis, but the titers of glucose, C1 and ADA were all normal. Although there was no bacterium in the CSF, the patient's electroencephalography finding was abnormal. We diagnosed his condition as viral meningoencephalitis and started treatment with antiviral agents. Blood chemistry showed serum sodium of 130 mEq/l and plasma osmolarity was reduced to 272 mOsm/kg, while urine osmolarity was high at 353 mOsm/kg. Two potential causes of hyponatremia in this patient were the syndrome of inappropriate secretion of antidiuretic hormone (SIADH) or cerebral salt wasting syndrome (CSWS). Physical findings revealed a contracted extracellular fluid volume, strongly suggesting the presence of CSWS. The massive urine sodium loss overcoming sodium intake supported this diagnosis. After treatment with vigorous sodium and volume replacement for over 4 weeks, hyponatremia as well as meningoencephalitis were improved without any complication. To the best of our knowledge, this is the first report on CSWS in a patient with viral meningoencephalitis. PMID- 17128885 TI - [An IgA nephropathy case with highly reduced urinary protein concomitant with reduced obesity]. AB - We reported a case of a 38-year-old woman with both massive proteinuria and severe obesity. We diagnosed her as metabolic syndrome from her waist size of over 90 cm around her umbilicus, hyperlipidemia (high TG level) and hypertension. The urinary protein was more than 3.5 g/day and body mass index was 38.7 at admission. The renal biopsy specimen revealed IgA nephropathy. According to Clinical guidelines of IgA nephropathy 2nd version, Committee of IgA Nephropathy the Special Study Group on Progressive Glomerular Disease, the Ministry of Health, Labor and Welfare of Japan-, her prognosis belonged to a rather poor group. We planed to administer steroid treatment first, however considering the adverse effects of steroid therapy, such as hyperlipidemia and diabetes mellitus, we tried to decrease her body weight as much as possible, and then treated her with both angiotensin converting enzyme inhibitor and anti-platelet drug. After her body mass index (body weight) was approximately 30.1 % (30 kg) less than that on admission, a parallel reduction of urinary protein was observed, and the final level was approximately 0.18 g/day. Decline in the body weight, diet and exercise were the chief measures that reduced the urinary protein without corticosteroid therapy. PMID- 17128886 TI - [A pregnant woman with nephrotic syndrome and focal segmental lesions]. AB - A 32-year-old woman was admitted at 36 weeks' gestation because of increasing proteinuria and generalized edema. At the time of admission, serum creatinine was 1.3 mg/dl, and urinalysis demonstrated 4+ protein and 2+ occult blood. During her pregnancy, her blood pressure had been in the normal range. A normal healthy female neonate was delivered by caesarean section at 38 weeks' gestation. After delivery, the woman's 24-hour urine protein excretion was 11 g/day and serum albumin was 1.4 g/dl , hence nephrotic syndrome was diagnosed. Eleven days after delivery, a renal biopsy showed focal segmental lesions with glomerular epithelial cell injury. She was given 50 mg/day prednisolone and after a month, her 24-hour urinary protein excretion decreased to 2 g/day. One year later, she achieved complete remission. Although she had a relapse of nephrotic syndrome after twenty-one months, steroid therapy again achieved a good response. PMID- 17128887 TI - Diffraction-enhanced imaging of the rat spine. AB - INTRODUCTION: Diffraction-enhanced imaging (DEI) uses monochromatic synchrotron X rays to image tissue. This technique has been shown to produce superior bony and soft tissue characterization when compared with conventional absorption radiography. Application of this imaging modality is under investigation, and this study represents the first DEI analysis of the vertebral column. METHODS: Four male Wistar rats were studied. Spine muscle blocks were imaged in 3 of the rats after thoracic laminectomy (n = 1), after lumbar laminectomy (n = 1), and in a control condition (n = 1). The fourth rat was imaged as a whole animal control. Conventional radiography and synchrotron-supported DEI at 40 keV were performed on all specimens. We compared images side by side, using a nonvalidated subjective assessment technique. RESULTS: DEI produced superior visualization of the vertebral anatomy, compared with conventional absorption radiography for all specimens. Greater bony and soft tissue detail was noted, with improved image contrast. In addition to imaging the anatomical structures, DEI showed the polyglactin suture material used for fascial closure in the 2 animals that underwent surgery. Artifact from air bubbles was present on DEI images but not on plain radiographs. CONCLUSIONS: This represents the first use of DEI, a novel imaging modality, to image the vertebral column. It provides excellent anatomic detail with superior contrast and visualization of both bone and soft tissue when compared with conventional radiography. Future applications of this investigational technique may include analysis of spinal fusion as well as degenerative and neoplastic conditions of the spine. PMID- 17128888 TI - An overview of digital compression of medical images: can we use lossy image compression in radiology? AB - The increasing volume of data generated by new imaging modalities such as multislice computed tomography scanners and magnetic resonance imaging justifies the use of lossy compression techniques to decrease the cost of storage and improve the efficiency of transmission over networks for teleradiology or for access to electronic patient records. We summarize here the most commonly used compression techniques and compare their main features. Having conducted an extensive literature review, we present a range of average compression ratios for different modalities and body parts. This article lays the groundwork for further evaluation with standardized statistical methods to ultimately elaborate acceptable compression guidelines. PMID- 17128889 TI - The clinical interventional radiologist: results of a national survey by the Canadian Interventional Radiology Association. AB - OBJECTIVE: To determine the level of clinical responsibility interventional radiologists in Canada currently have within their practice and would like to have within their future practices. METHODS: An anonymous online survey was e mailed to all members of the Canadian Interventional Radiology Association. The survey was open for a period of 2 months. RESULTS: A total of 75 surveys were received, of a possible 247, a response rate of 30.4%. Responses regarding general measures of clinical duties were collected. The current situation in Canada is mixed, in that while most (82%) respondents perform procedures that require an overnight admission, only 11% have a dedicated interventional radiology (IR) ward and 29% have admitting privileges. Most (73%) respondents stated that interventional radiologists in Canada should become more clinical. The most common reason cited for a lack of admitting privileges was a lack of time (44%), followed by a lack of hospital or administrative support (40%), "other" (20%), and inadequate remuneration (14%). CONCLUSIONS: Most respondents believe that interventional radiologists should become more clinically oriented. The most frequently noted obstacles to becoming more clinically oriented are reluctant administration, lack of time, and inadequate remuneration for clinical duties. PMID- 17128890 TI - A rural CT scanner: evaluating the effect on local health care. AB - OBJECTIVE: The first small rural hospital in Ontario to propose a computed tomography (CT) scanner was in Walkerton, a town 160 km north of London. The Ontario Ministry of Health approved the proposal as a pilot project to evaluate the effect on local health care of a rural scanner. This evaluation study had 3 parts: a survey of physicians, a survey of patients, and an analysis of population CT scanning rates. METHOD: The physicians in the area served by the scanner were asked about its impact on their care of their patients in a mailed questionnaire and in semistructured interviews. Scanner outpatients were given a questionnaire in which they rated the importance of its advantages. The analysis of scanning rates--the ratio of number of scans to estimated population--compared rates in the area with other Ontario rates before and after the scanner was introduced. RESULTS: The physicians reported that local CT allowed them to diagnose and treat patients sooner, closer to home, and with greater confidence. On average, 75% of the patients ranked faster and closer access as very important. Scanning rates in the area rose, although they did not match urban rates. CONCLUSIONS: The study confirms that the rural scanner changed the area's health care in significant ways and that it helped to narrow the gap between rural and urban service levels. We recommend that CT be expanded to other rural regions. PMID- 17128891 TI - In-training evaluation using hand-held computerized clinical work sampling strategies in radiology residency. AB - PURPOSE: The in-training evaluation and final in-training evaluation are the mainstay format for evaluation summaries in Canadian residency training programs. This study investigates the feasibility of a clinical work sampling (CWS) approach to evaluation in radiology residency, with the aid of personal hand-held computing devices METHODS: This study was conducted over a 1-year period with 14 radiology residents spanning 4 postgraduate years. Residents were provided with a hand-held device to enter evaluation data, with entries assessing 9 categories of resident performance. Results from the CWS entries were compared with standard in training evaluations completed at the end of the residents' rotations, as well as with an established annual objective evaluation tool. RESULTS: The overall reliability of the CWS approach according to the observed 7 forms per resident was 0.62, suggesting that a minimum of 20 forms would be required to achieve a reliability of 0.80. For the in-training evaluation report (ITER), internal consistency was 0.98, reflecting very high correlations between categories and indicating that the individual categories are not discriminating. Correlation across rotations was 0.36, which is low for summative evaluation. Correlation between the 2 measures was 0.47 (P = 0.09); neither measure was correlated with the American College of Radiology evaluation. CONCLUSION: The CWS strategy is feasible for adaptation to radiology residency, although compliance with voluntary entries was less than expected. It is not clear whether this reflects the additional burden of using the hand-held device, the fact that entries were voluntary rather than mandatory, or the many demands on both residents and evaluators. The added potential of this evaluation format includes the opportunity to discuss performance at the time of data entry, rather than resorting to the usual end-of-rotation evaluation. Nevertheless, the study has shown that the ITER remains only of marginal value for summative evaluation; the addition of the CWS would require at least 20 forms for acceptable reliability and might not justify the additional cost and complexity. PMID- 17128892 TI - Magnetic resonance imaging of thoracic aortic pseudoaneurysm. PMID- 17128893 TI - Answer to case of the month #111: Cleidocranial dysostosis. PMID- 17128894 TI - Answer to case of the month #112: Progressive multifocal leukoencephalopathy with characteristic magnetic resonance imaging and magnetic resonance spectroscopy findings. PMID- 17128895 TI - Invest in the future--become a mentor. PMID- 17128896 TI - Functional decline in hospitalized elders. PMID- 17128897 TI - The challenges of shift work. PMID- 17128898 TI - Venous thromboembolism (VTE) prophylaxis: implications for medical-surgical nurses. AB - Nurses are responsible for continual patient evaluation as they address patient needs, provide essential input into evaluation and risk stratification, and make recommendations regarding prevention-oriented strategies (American Health Consultants, 2002a). As a result, nurses are in a pivotal role to prevent VTE. Education about all aspects of VTE prevention should be included in basic nursing programs and must continue throughout a nurse's career through continuing education, reading, and dissemination of information from current health care literature. PMID- 17128899 TI - Glass ampules and filter needles: an example of implementing the sixth 'r' in medication administration. PMID- 17128900 TI - Surgical wound dehiscence. AB - Despite advances in preoperative care, the rate of surgical wound dehiscence has not decreased in recent years; 1%-3% of patients experience wound dehiscence. A nursing goal for the postoperative patient is always prevention of wound dehiscence. Recognition of risk factors is essential. For example, older males with ascites are at very high risk. Prevention of wound infection and mechanical stress on the incision are important. Management of dehisced wounds may include immediate surgery if bowel is protruding from the wound. If surgery is not needed, management is essentially the same as that of any other wound through maintenance of a moist wound environment, reduction of bioburden and pain, and promotion of granulation tissue. Mortality rates associated with dehiscence have been reported between 14% and 50% (Hanif et al., 2000; Waqar et al., 2005). One of the complications (morbidity) of dehisced wounds is an incisional hernia, which develops in an estimated 43% of patients (van't et al., 2004). Researchers followed 126 patients who had wound dehiscence repair for a mean of 37 months and found that 31% of the hernias were diagnosed more than 2 years postoperatively. Nurses need to ensure adequate nutrition and reduced tension on the abdomen, and implement measures to prevent incisional infections. PMID- 17128901 TI - Secondary lymphedema in the cancer patient. PMID- 17128903 TI - Avoiding restraints in patients with dementia. PMID- 17128902 TI - Continuing education changes affect MEDSURG Nursing. PMID- 17128904 TI - Bladder ultrasound. PMID- 17128905 TI - Authentic leadership. AB - There is leadership, and then there is authentic leadership. If you are not willing to engage from your heart, to passionately work to create a greater quality of work life for front-line staff every day, and to push yourself to the ultimate limit to make that happen, you might be a leader, but you will not be perceived as an authentic leader. Authentic leaders love, challenge people to do what they didn't believe was possible, and generate the energy to make the impossible possible by their passion for their people, their patients, and for doing the right thing. Thankfully, there are leaders who are willing to live on the edge, model their love, and inspire people to change the world. Will you be one of them? PMID- 17128906 TI - Community petitions: threshold options revealed in DoH paper. PMID- 17128907 TI - NPSA 'struggling' with adverse incident reports. PMID- 17128908 TI - Efficiency savings costed 2.2bn pounds sterling. PMID- 17128909 TI - Time to give PECs the muscle to make a real difference. PMID- 17128910 TI - On disinvestment and decluttering. PMID- 17128911 TI - Why the bedside has a place in the boardroom. PMID- 17128912 TI - Barometer. Acute trusts Oct 2006. PMID- 17128913 TI - Mental health. Out of harm's way. AB - There are around 350 psychiatric intensive care units but standards vary. Guidelines have been published, bu there is no obligation to follow them. A number of units are inappropriately equipped--for example, 35 per cent lack single-sex facilities. Many PICU beds are blocked by patients who should be treated in low-secure units. PMID- 17128914 TI - Productive ward. Stand by your bed. PMID- 17128915 TI - [Late stent thromboses a neglected problem? Indications of increased risk with drug-eluting stents]. PMID- 17128916 TI - [Drug-eluting stents can increase the risk of late stent thrombosis]. PMID- 17128917 TI - [The Chlamydia infection epidemic threatens women's reproductive health. A longitudinal study points out the need of effective strategy to break the trend]. PMID- 17128918 TI - [Psychological first aid after disasters and accidents]. PMID- 17128919 TI - [Interactions between genes and environment predict criminality, depression and alcohol dependence]. PMID- 17128920 TI - [Sex--only for the rich and healthy? PDE5 inhibitors too expensive for low income men]. PMID- 17128921 TI - [Reduced incidence of endophthalmitis following cataract extraction. National registration has found the risk factors]. PMID- 17128922 TI - [Syncope]. PMID- 17128923 TI - [Functional imaging medicine--a new specialty with great prospects]. PMID- 17128924 TI - [Psychiatry not mature for compulsory outpatient care]. PMID- 17128925 TI - [If approved applications are displayed on the Internet--there is no need for a new trials directory]. PMID- 17128926 TI - [The penal code principles can not be underestimated!]. PMID- 17128927 TI - [Patients unable to pay are detained in hospitals]. PMID- 17128929 TI - [Norwegian medical education a step towards the university in Gothenburg?]. PMID- 17128930 TI - Bone microstructure and its hidden information. AB - Human bone micromorphology gives clues to a variety of life history parameters, such as individual age, health status, and physical activity. In the course of an ongoing study, thin cross sections of femoral compact bone from three skeletal series are investigated for different purposes. The first series consists of 103 adult skeletons excavated from a 19th century hospital graveyard in Basel, Switzerland. Several disease- and stress-markers, like layers of arrested growth or other conspicuous microstructural composition were observed. Another 36 individuals come from the dissection room of the Ludwig Maximilians University of Munich. These individuals have an average age at death of about 80 years and offer the possibility to investigate the micromorphological characteristics of individuals of very advanced age. Finally, 72 medieval subadult skeletons shall serve for the establishment of a relationship between individual age and bone microstructural parameters according to the different ontogenetic stages. PMID- 17128931 TI - [Degradation of biomolecules in bones: effects of the biological forensics as an example of the stability of isotope ratios in collagen]. AB - Modern bone samples were experimentally degraded by incubation into water at increased temperature and examined in terms of their collagen content, the stable C and N isotopic ratios, and the molar C/N ratio. The same analyses were carried out with archaeological human bone of varying age (300 up to 8000 years). The experimentally degraded samples exhibited changes of the collagen's integrity, which influence the stable isotope ratios. In the case of the archaeological material, a correlation between stable delta13C- and delta15N-values and collagen content could be demonstrated. The molar C:N ratio was no suitable criterion for the assessment of the state of preservation of extractable collagen. PMID- 17128932 TI - Reconstruction of an aquatic food web: Viking Haithabu vs. Medieval Schleswig. AB - In the last two decades, the analysis of stable isotopes of carbonate and collagen from archaeological bone finds became a useful tool in the reconstruction of ancient food webs. Nevertheless there is still only little information available about aquatic food webs, in particular concerning brackish water ecosystems. The Schlei Fjord in Schleswig-Holstein, Germany, is such an ecosystem and was investigated to determine stable isotopic values for nitrogen, carbon and oxygen from bone collagen and carbonate in archaeological bone finds. Wild birds, mammals and fish bones from the Viking settlement Haithabu and the medieval town of Schleswig have been analyzed in this study to determine stable isotope values for marine and limnic species and to investigate possible isotopic gradients for mixing fresh water and salt water ecosystems. PMID- 17128933 TI - Social structures and social relations--an archaeological and anthropological examination of three early Medieval separate burial sites in Bavaria. AB - As part of an interdisciplinary cooperation, three early medieval separated burial sites from central Bavaria were subjected to archaeological and anthropological examinations. Separated burial sites are probably an expression of social stratification. This paper focuses on two characteristic aspects of these separated burial sites: (1) structure of the site, and (2) biological manifestation of a social upper class. The separated burial grounds Etting/Sandfeld (31 individuals), Grossmehring (44 individuals) and Kelheim (43 individuals), all located in southern Bavaria, were analyzed. Sex and age at death of all individuals were determined, and dietary behaviour was reconstructed by means of carbon and nitrogen isotope ratios in bone collagen. Local conditions such as climate, soil conditions or the intensity of agriculture can provoke significant variations in plant delta15N-values, which could lead to a shift of the baseline values of the corresponding trophic web. To facilitate the comparison of isotopic data from different sites, delta15N values of cattle bones were taken as a reference for the human data (presuming the diet to consumer chain). The results of dietary reconstruction indicate that the populations enjoyed very good living conditions with a primarily animal protein-based diet. Furthermore, the isotope analysis revealed more detailed indications for certain individuals regarding their social status. Some individuals can even be appointed as possible chiefs of the population, since anthropological and archaeological interpretations were in total agreement: It was possible to identify persons of a higher social status based on the structure of the burial site, grave goods and the isotopic analysis. PMID- 17128934 TI - Class endogamy, inbreeding and migration during the Argentinean colonial period: analysis based on individuals of European ancestry. AB - During the Spanish colonial period, Cordoba was an important town due to its location suitable for controlling communication throughout a wide region of Argentina. As a typical colonial society, where individuals belonged to a well established social class, marriages among Whites were the result of strict norms and prejudices founded on nets of kinship, friendship and neighbouring. The objective of this paper is to determine, whether the elevated class endogamy among Whites during the colonial period resulted in high consanguinity and whether a process of selection of surnames was present in 1813. In both cases the interaction with the migratory flow was taken into account. It is concluded that (a) Although an endogamy of "class" existed among Whites, there is no evidence of the consequences on the biological structure of the population through preferential mating between relatives, as deduced from the low level of inbreeding from isonymy. (b) There was no differential reproduction reflecting the selection of families carrying certain surnames; therefore surnames followed a "neutral" model. (c) The diversity of surnames and the gene flow deduced from them were of a magnitude similar to present populations; this fact could have balanced the effect of the "class" endogamy. (d) Finally, the congruence between information derived from the population age distribution and historic data and the quantification of migration from surnames allowed the detection of mistakes in the census data file regarding the number of foreigners, thus validating the use of surnames as biological markers of the population structure. PMID- 17128935 TI - Body composition, weight status, body image and weight control practices among female adolescents from eastern Austria. AB - The prevalence rates of obesity, but also of altered eating behaviour up to clinically manifested eating disorders have increased dramatically during the last years. In the present study the weight status, body composition and eating behaviour as well as weight controlling strategies of 1752 female adolescents ageing between 12 and 18 years (x=14.6) from Eastern Austria were documented. Furthermore, data regarding socioeconomic status and educational level of the probands were collected. It could be shown, that a higher educational level or a higher socioeconomic status was associated with a significantly lower fat percentage and a significantly lower prevalence of overweight and obesity. Underweight and severe underweight was predominantly found among girls ageing between 15 and 18 years of higher educational level. Weight controlling behaviour such as stepping on scale and altered or restricted eating behaviour was widespread among the probands, independent of weight status and socioeconomic parameters. Body weight and shape concerns were also found very frequently independent of educational level. The results of the present study corroborate the findings of former investigations, that disturbed eating behaviour and concerns regarding body weight and shape are found among all socioeconomic groups. PMID- 17128936 TI - Overweight and obesity prevalence in urban Sardinian children. AB - The aim of this study is to evaluate the prevalence of overweight and obesity in children (6-10 years) of the city of Cagliari (Sardinia, Italy) with different socioeconomic status. The sample is composed by 1000 children, 500 males and 500 females, 6 to 10 years old, attending primary schools in Cagliari during 2003. For uniformity with similar Italian studies, in this study overweight and obesity were defined as calculated from the charts published by Tanner et al. (1966). In the Cagliari children, the prevalence of obesity is 22.70%. The percentages of overweight and obese children increase with age: respectively from 11.5% and 14.0% at 6 years to 15.4% and 22.7% at 10 years. There are higher numbers of overweight and obese boys than girls in all the age classes. Both males and females show an increasing percentage of overweight and obesity as the socioeconomic level decreases. Males present higher percentages of overweight and obesity than females of the same social level, i.e. in the lowest social category overweight is 18.68% in males and 13.60% in females and obesity 26.46% in males and 23.62% in females. The standard multivariate regression analysis with the indicator of overweight and obesity as dependent variable showed that the sex (male), socio-economic status, maternal schooling and sums of the limb and trunk skinfolds have the greatest influence on overweight and obesity. The results suggest that overweight and obesity are becoming serious social and health problems in Sardinia. PMID- 17128937 TI - Q-factor analysis as a tool for phylogenetic studies of morphometric data. AB - The statistical procedure of Q-factor analysis was applied with the aim to detect complex morphological characters, which clearly separate taxonomic groups. Since Q-factor analysis, unlike cluster analysis, does not prescribe a hierarchical grouping, the results can be interpreted phylogenetically using an outgroup comparison. The essential steps of the approach are demonstrated by an empirical study on the phylogeny of Callitrichinae. Except for some complications concerning the determination of the number of relevant factors and factor rotation, the procedure proved to be a suitable instrument for phylogenetic research. PMID- 17128939 TI - 4 steps to data mining. PMID- 17128938 TI - [Art history anthropology: a new interdisciplinary overlap using the examples of Caravaggio and Mozart]. AB - The established identification of human faces on images may also be applied to paintings. This demands an extension of the known methodology: A check is performed if the depicted trait expressions may occur in nature at all or if they are an "artefact" of artistic freedom. Two cases are presented here, which illustrate an identification and an identity exclusion. PMID- 17128940 TI - Finding gold in your denial database. PMID- 17128941 TI - What to do with the paper? PMID- 17128942 TI - A real Somers storm. PMID- 17128943 TI - Kidney Failure? Dialysis is the only medical service without automatic inflation updates from Medicare. Some people are trying to change that. AB - According to Robert Provenzano, left, president of the Renal Physicians Association, just because dialysis providers have shown strong profits does not mean an automatic update for Medicare reimbursement for dialysis isn't necessary. Unlike other medical products and services, which saw payment increases of 107% between 1990 and 2006, the increase in dialysis payments didn't even hit 7% during that period. PMID- 17128944 TI - CMS extends deadline, but some say '09 still too soon for quality measures. PMID- 17128945 TI - Red cross revamps. Charity to cut size of board, realign responsibilities. PMID- 17128946 TI - Hillenbrand ponders split. PMID- 17128947 TI - For their eyes only. HHS keeping privacy meetings under wraps. PMID- 17128948 TI - Misys' panel raises questions. Advisers could be subjected to IT conflicts of interest. PMID- 17128949 TI - You must remember this...seniors shouldn't be shocked! Shocked! To learn perils of Medicare managed care. PMID- 17128950 TI - Heart-healthy. Staff commitment, use of technology, attention to detail help facilities land on Solucient's list of 100 top cardiac hospitals. PMID- 17128951 TI - Joined at the hip. New partnership revives Okla. orthopedic hospital. PMID- 17128952 TI - Largest master's programs in health administration ranked by total number of students enrolled, based on 2005-06 survey of programs. PMID- 17128953 TI - Straight talk new approaches to healthcare. The healthcare quality equation. AB - Though various health organizations may define and approach quality differently, the common denominator is the patient's perspective, experience and outcome. Quality issues should be an important part of health system executives' agenda. Successful quality programs have a number of key ingredients, including decision reporting tools and business analytics information--which depend on robust information reporting and a system-wide electronic medical record--as well as highly motivated employees and physicians. In this installment of Straight Talk, representatives from two health systems--Kaiser Permanente and Clarian Health Partners--discuss their quality initiatives. PMID- 17128954 TI - Alcohol, neurotransmitter systems, and behavior. AB - Alcohol affects several neurotransmitter systems within the brain. In this article, the author describes its effects on 5 major ones: glutamate, gamma-amino butyric acid (GABA), dopamine, serotonin, and opioid systems. The author also notes the interactions and interdependencies of these transmitters, and provides details on both immediate effects and long-term adaptations. Last, the author explains several psychopharmacological treatments for alcoholism and the effects of these treatments on transmitters, and draws conclusions. PMID- 17128955 TI - Impact of the hormonal milieu on the neurobiology of alcohol dependence and withdrawal. AB - Alcoholism, or alcohol dependence, is a complex disorder with withdrawal symptoms that are often problematic for those trying to recover from their dependence. As researchers attempt to elucidate the neurobiological underpinnings of alcohol dependence and withdrawal, it is becoming clear that numerous factors, including the hormonal environment, impact the manifestations of this disorder. Of particular interest is the observation that women have fewer and less severe withdrawal symptoms than do men even though they tend to suffer greater physiological harm from excessive alcohol consumption. In this article, the authors present an overview of their understanding of how gonadal and stress hormones interact with alcohol, which results in differential neurobiological responses between males and females. Thus far, data generated from representative animal models have shown significant differences between the sexes in behavioral responses and neuroadaptations to chronic alcohol consumption and withdrawal. Accumulating evidence suggests that treatment of alcoholism, including withdrawal, should be tailored to the patient's gender and hormonal status. PMID- 17128956 TI - Possible contributors to the gender differences in alcohol use and problems. AB - Research on alcohol use and problems has demonstrated a much higher rate of alcohol use disorders (AUDs) among men compared with women. The authors review the most frequently researched biological and psychosocial factors that may play a role in the gender differences in alcohol use and problems. Among the biological factors, women might carry a lower genetic risk for AUDs and tend to suffer more negative biological consequences from drinking as compared with men. Regarding psychosocial factors, men appear to be more likely than women to manifest certain risk factors for alcohol use and problems (e.g., fewer perceived social sanctions for drinking, positive expectancies for alcohol use, personality traits such as impulsivity) and have fewer protective factors. Although these findings have helped to explain some of the gender differences in alcohol use and problems, there are inconsistencies in the literature. Additionally, many studie may not have enough statistical power to detect gender differences. The authors suggest that the biological and social consequences of alcohol abuse for women may be especially potent protective factors against AUDs and discuss the possibility of public health campaigns developed on the basis of this idea. PMID- 17128957 TI - Tobacco use, alcohol dependence, and cognitive performance. AB - Chronic alcohol abuse has long been associated with a mild, generalized pattern of cognitive decrements. However, it is important to note that problem drinking rarely occurs in isolation from abuse of other drugs. For people dependent upon alcohol, tobacco is one of the mostly commonly coabused substances. Recent research suggests that individuals with alcohol dependency may gravitate toward tobacco use, in part, because of the positive effects of nicotine on aspects of cognitive performance that may be compromised as a consequence of chronic alcohol misuse. In this article, the author focuses on the effects of nicotine on behavioral and electrophysiological indexes of cognitive performance, and the impact of these effects on alcohol-related cognitive decrements. The author discusses implications of these findings in the context of treatment and recovery of people with alcoholism. PMID- 17128958 TI - Perceptions of one's attacker's intentions following an aggressive interaction involving alcohol. AB - The authors investigated and assessed the perceptions of one's attacker's intentions following an aggressive interaction involving alcohol. Participants were 328 (163 men and 165 women) healthy social drinkers between 21 and 35 years of age. After participants had consumed a beverage containing either alcohol or a placebo, the authors tested them on a modified version of the Taylor Aggression Paradigm (TAP; S. Taylor, 1967) in which participants received mild electric shocks from, and administered shocks to, a fictitious opponent (the attacker) under the guise of a competitive task. Aggressive behavior was operationalized as the shock intensities administered to the fictitious opponent. The authors queried participants about their perceptions of their opponents' intentions toward them on the task. Overall, participants who were least aggressive on the TAP perceived their opponent to have the most aggressive intentions and those who were the most aggressive perceived their opponent to have the least aggressive intentions. Alcohol only seemed to play a role for women. It appeared to decrease aggressive perceptions for the least aggressive women and to increase such perceptions for the most aggressive women. The authors discuss results according to L. Huesmann's (1988) cognitive script model of aggression. PMID- 17128959 TI - Heavy drinking in college students: who is at risk and what is being done about it? AB - Problem drinking and related consequences are a major social issue plaguing college campuses across the United States. Each year, alcohol is responsible for fatalities, assaults, serious injuries, and arrests that occur among college students. The authors review and discuss the risk factors, drinking patterns, and consequences that are relevant to the general student population. In addition, the authors highlight individuals at an increased risk of experiencing alcohol related problems, such as Greek-letter social organization members and student athletes. The authors also discuss the interventions that attempt to reduce risky drinking and related problems in these subgroups as well as the future directions for research. PMID- 17128960 TI - Help seeking for alcohol problems in a college population. AB - Despite the availability of effective treatments for problem drinkers, only a minority seek professional assistance. The authors investigated the influences on help seeking for alcohol-related difficulties in two separate studies. In Study 1, 59 men and 74 women completed questionnaires regarding their alcohol use, related problems, readiness to change, and help-seeking interest. The authors also developed a measure of stigma related to alcohol help seeking. Problem recognition, anxiety symptoms, and stigma predicted a global rating of help seeking. Students (22 men and 60 women) participated in Study 2, which incorporated a multi-item measure of help-seeking intentions as well as attitude and personality measures. Positive attitudes, less perceived stigma, and greater self-efficacy increased help seeking. The findings highlight the importance of both approach and avoidance factors that affect help-seeking decisions, and the authors suggest directions for further research. PMID- 17128961 TI - Native Americans and alcohol: past, present, and future. AB - Native Americans have higher rates of alcohol use, frequency of use, and increased rates of fetal alcohol syndrome, compared with other ethnic groups (J. Hisnanick, 1992; P. A. May, 1996; J. M. Wallace et al., 2003). High prevalence rates of alcohol misuse among Native Americans must be understood in light of their unique history, which has resulted in trauma and exposure to many risk factors for problem alcohol use. Many risk factors have been identified in the general population; however, only some of these risk factors have been examined among Native American populations. The unique history and world view of Native Americans mean that, often, risk factors operate differently from the way they do in other populations. The authors discuss interventions and promising treatments. PMID- 17128962 TI - Alcohol use among older adults. AB - Alcohol use problems among older adults have been called the "invisible epidemic." As the population of older adults continues to grow, there is an increased need to reexamine alcohol use in this population. The authors provide an overview on alcohol use in the over-60 age group. The main areas of focus included research on the prevalence of drinking in that population, as well as comments on the best practices in assessment and psychological treatment. Several screening assessments have been recommended for use with older adults, such as the CAGE questionnaire, Michigan Alcohol Screening Test-Geriatric version, Alcohol-Related Problems Survey, and the Alcohol Use Disorders Identification Test. The authors note age-appropriate psychological treatment interventions that include brief interventions, family interventions, motivational counseling, and cognitive behavioral therapies. Barriers to assessment and treatment are also discussed. PMID- 17128964 TI - Resilience of Rhodobacter sphaeroides cytochrome bc1 to heme c1 ligation changes. AB - Typically, c hemes are bound to the protein through two thioether bonds to cysteines and two axial ligands to the heme iron. In high-potential class I c type cytochromes, these axial ligands are commonly His-Met. A change in this methionine axial ligand is often correlated with a dramatic drop in the heme redox potential and loss of function. Here we describe a bacterial cytochrome c with an unusual tolerance to the alternations in the heme ligation pattern. Substitution of the heme ligating methionine (M185) in cytochrome c1 of the Rhodobacter sphaeroides cytochrome bc1 complex with Lys and Leu lowers the redox midpoint potential but not enough to prevent physiologically competent electron transfer in these fully functional variants. Only when Met-185 is replaced with His is the drop in the redox potential sufficiently large to cause cytochrome bc1 electron transfer chain failure. Functional mutants preserve the structural integrity of the heme crevice: only the nonfunctional His variant allows carbon monoxide to bind to reduced heme, indicating a significant opening of the heme environment. This range of cytochrome c1 ligand mutants exposes both the relative resilience to sixth axial ligand change and the ultimate thermodynamic limits of operation of the cofactor chains in cytochrome bc1. PMID- 17128963 TI - Calmodulin, conformational states, and calcium signaling. A single-molecule perspective. AB - Single-molecule fluorescence measurements can provide a new perspective on the conformations, dynamics, and interactions of proteins. Recent examples are described illustrating the application of single-molecule fluorescence spectroscopy to calcium signaling proteins with an emphasis on the new information available in single-molecule fluorescence burst measurements, resonance energy transfer, and polarization modulation methods. Calcium signaling pathways are crucial in many cellular processes. The calcium binding protein calmodulin (CaM) serves as a molecular switch to regulate a network of calcium signaling pathways. Single-molecule spectroscopic methods can yield insights into conformations and dynamics of CaM and CaM-regulated proteins. Examples include studies of the conformations and dynamics of CaM, binding of target peptides, and interaction with the plasma-membrane Ca2+ pump. Single-molecule resonance energy transfer measurements revealed conformational substates of CaM, and single molecule polarization modulation spectroscopy was used to probe interactions between CaM and the plasma-membrane Ca2+-ATPase. PMID- 17128965 TI - Mechanistic characterization of N-formimino-L-glutamate iminohydrolase from Pseudomonas aeruginosa. AB - N-Formimino-l-glutamate iminohydrolase (HutF) from Pseudomonas aeruginosa catalyzes the deimination of N-formimino-l-glutamate in the histidine degradation pathway. An amino acid sequence alignment between HutF and members of the amidohydrolase superfamily containing mononuclear metal centers indicated that residues Glu-235, His-269, and Asp-320 are involved in substrate binding and activation of the nucleophilic water molecule. The purified enzyme contained up to one equivalent of zinc. The metal was removed by dialysis against the metal chelator dipicolinate with the complete loss of catalytic activity. Enzymatic activity was restored by incubation of the apoprotein with Zn2+, Cd2+, Ni2+, or Cu2+. The mutation of Glu-235, His-269, or Asp-320 resulted in the diminution of catalytic activity by two to six orders of magnitude. Bell-shaped profiles were observed for kcat and kcat/Km as a function of pH. The pKa of the group that must be unprotonated for catalytic activity was consistent with the ionization of His 269. This residue is proposed to function as a general base in the abstraction of a proton from the metal-bound water molecule. In the proposed catalytic mechanism, the reaction is initiated by the abstraction of a proton from the metal-bound water molecule by the side chain imidazole of His-269 to generate a tetrahedral intermediate of the substrate. The collapse of the tetrahedral intermediate commences with the abstraction of a second proton via the side chain carboxylate of Asp-320. The C-N bond of the substrate is subsequently cleaved with proton transfer from His-269 to form ammonia and the N-formyl product. The postulated role of the invariant Glu-235 is to ion pair with the positively charged formimino group of the substrate. PMID- 17128966 TI - Fatty acid-specific fluorescent probes and their use in resolving mixtures of unbound free fatty acids in equilibrium with albumin. AB - We report the first measurements for profiling mixtures of unbound free fatty acids. Measurements utilized fluorescent probes with distinctly different response profiles for different free fatty acids (FFA). These probes were constructed by labeling site-specific mutants of the rat intestinal fatty acid binding protein (rI-FABP) with acrylodan. The probes were produced and screened by high-throughput methods, and from more than 30 000 such probes we selected six that together have sufficient specificity and sensitivity for resolving the profile of unbound FFA (FFAu) in mixtures of different FFAu. We developed analytical methods to determine the FFAu profile from the fluorescence (ratio) response of the different probes and used these methods to determine FFAu profiles for mixtures of arachidonate, linoleate, oleate, palmitate, and stearate in equilibrium with bovine serum albumin (BSA). Measurements were performed using mixtures with a range of total FFAu concentrations, including 0.9 nM, which is similar to normal plasma levels. We also measured single FFA binding isotherms for BSA and found that binding was described well by six to seven sites with the same binding constants (Kd). The Kd values for the FFA (4-38 nM) were inversely related to the aqueous solubility of the FFA. We constructed a model with these parameters to predict the FFAu profile in equilibrium with BSA and found excellent agreement between the profiles measured using the FFA probes and those calculated with this model. These results should lead to a better understanding of albumin's role in buffering FFAu and to profiling FFAu in intra- and extracellular biological fluids. PMID- 17128967 TI - Resonance Raman study of ferric heme adducts of dehaloperoxidase from Amphitrite ornata. AB - The study of axial ligation by anionic ligands to ferric heme iron by resonance Raman spectroscopy provides a basis for comparison of the intrinsic electron donor ability of the proximal histidine in horse heart myoglobin (HHMb), dehaloperoxidase (DHP), and horseradish peroxidase (HRP). DHP is a dimeric hemoglobin (Hb) originally isolated from the terebellid polychaete Amphitrite ornata. The monomers are structurally related to Mb and yet DHP has a peroxidase function. The core size marker modes, v2 and v3, were observed using Soret excitation, and DHP-X was compared to HHMb-X for the ligand series X = F, Cl, Br, SCN, OH, N3, and CN. Special attention was paid to the hydroxide adduct, which is also formed during the catalytic cycle of peroxidases. The Fe-OH stretching frequency was observed and confirmed by deuteration and is higher in DHP than in HHMb. The population of high-spin states of the heme iron in DHP was determined to be intermediate between HHMb and HRP. The data provide the first direct measurement of the effect of axial ligation on the heme iron in DHP. The Raman data support a modified charge relay in DHP, in which a strongly hydrogen-bonded backbone carbonyl (>C=O) polarizes the proximal histidine. The charge relay mechanism by backbone carbonyl >C=O-His-Fe is the analogue of the Asp-His-Fe of peroxidases and Glu-His-Fe of flavohemoglobins. PMID- 17128968 TI - The plasticins: membrane adsorption, lipid disorders, and biological activity. AB - The present study investigates the relationships between structural polymorphism, adsorption onto membrane mimetic support, lipid disturbance, and biological activity of bactericidal 23-residue, glycine-leucine-rich dermaseptin orthologues from the Phyllomedusinae frog skin, the "plasticins". Biological activities were evaluated using the membrane models DMPG (1,2-dimyristoyl-sn-glycero-3 phosphatidylglycerol) for prokaryotic membranes and DMPC (1,2-dimyristoyl-sn glycero-3-phosphatidylcholine) for eukaryotic membranes. We performed a conformational analysis of plasticins by molecular simulations and spectroscopic methods and analyzed phospholipid perturbations by infrared spectroscopy. Adsorption onto synthetic model membranes was quantified by surface plasmon resonance. Biological assays including antimicrobial and membrane potential dissipating activities, together with hemolytic tests and imaging analysis of cytotoxicity, were carried out to clarify the peptide-membrane interactions. Two major groups were distinguished: (i) Neutral plasticins revealed the presence of strong beta-structures with the zwitterionic or anionic phospholipid vesicles. They were weakly adsorbed in the range of antibacterial activity concentrations (micromolar). Nevertheless, for millimolar concentrations, they caused perturbations at the interface peptide-DMPG vesicles and in the bilayer alkyl chains, suggesting insertion into bacterial membranes. (ii) Cationic plasticins revealed multiple conformational transitions, including destabilized helix states, beta-structures, and disordered states. Peptide-lipid complex densities depended on hydrophobic bond strengths. The most soluble cationic plasticins were strongly adsorbed, with stable peptide-lipid interactions inducing noticeable perturbations of bilayer alkyl chains, pointing out possible insertion into bacterial membranes. In contrast, cytotoxic plasticins were less adsorbed, with less stable peptide-lipid interactions causing membrane dehydration, formation of peptide-membrane hydrogen bonds, and little disturbances of lipid alkyl chains. These characteristics could be compatible with their putative action on intracellular targets leading to apoptosis. PMID- 17128969 TI - Transmembrane helix-helix interactions: comparative simulations of the glycophorin a dimer. AB - The glycophorin helix dimer is a paradigm for the exploration of helix-helix interactions in integral membrane proteins. Two NMR structures of the dimer are known, one in a detergent micelle and one in a lipid bilayer. Multiple (4 x 50 ns) molecular dynamics simulations starting from each of the two NMR structures, with each structure in either a dodecyl phosphocholine (DPC) micelle or a dimyristoyl phosphatidylcholine (DMPC) bilayer, have been used to explore the conformational dynamics of the helix dimer. Analysis of the helix-helix interaction, mediated by the GxxxG sequence motif, suggests convergence of the simulations to a common model. This is closer to the NMR structure determined in a bilayer than to micelle structure. The stable dimer interface in the final simulation model is characterized by (i) Gly/Gly packing and (ii) Thr/Thr interhelix H-bonds. These results demonstrate the ability of extended molecular dynamics simulations in a lipid bilayer environment to refine membrane protein structures or models derived from experimental data obtained in protein/detergent micelles. PMID- 17128970 TI - Calcium binding to calmodulin mutants having domain-specific effects on the regulation of ion channels. AB - Calmodulin (CaM) is an essential, eukaryotic protein comprised of two highly homologous domains (N and C). CaM binds four calcium ions cooperatively, regulating a wide array of target proteins. A genetic screen of Paramecia by Kung [Kung, C. et al. (1992) Cell Calcium 13, 413-425] demonstrated that the domains of CaM have separable physiological roles: "under-reactive" mutations affecting calcium-dependent sodium currents mapped to the N-domain, while "over-reactive" mutations affecting calcium-dependent potassium currents localized to the C domain of CaM. To determine whether and how these mutations affected intrinsic calcium-binding properties of CaM domains, phenylalanine fluorescence was used to monitor calcium binding to sites I and II (N-domain) and tyrosine fluorescence was used to monitor sites III and IV (C-domain). To explore interdomain interactions, binding properties of each full-length mutant were compared to those of its corresponding domain fragments. The calcium-binding properties of six under-reactive mutants (V35I/D50N, G40E, G40E/D50N, D50G, E54K, and G59S) and one over-reactive mutant (M145V) were indistinguishable from those of wild-type CaM, despite their deleterious physiological effects on ion-channel regulation. Four over-reactive mutants (D95G, S101F, E104K, and H135R) significantly decreased the calcium affinity of the C-domain. Of these, one (E104K) also increased the calcium affinity of the N-domain, demonstrating that the magnitude and direction of wild-type interdomain coupling had been perturbed. This suggests that, while some of these mutations alter calcium-binding directly, others probably alter CaM-channel association or calcium-triggered conformational change in the context of a ternary complex with the affected ion channel. PMID- 17128971 TI - Structure of Arabidopsis thaliana At1g77540 protein, a minimal acetyltransferase from the COG2388 family. AB - We describe X-ray crystal and NMR solution structures of the protein coded for by Arabidopsis thaliana gene At1g77540.1 (At1g77540). The crystal structure was determined to 1.15 A with an R factor of 14.9% (Rfree = 17.0%) by multiple wavelength anomalous diffraction using sodium bromide derivatized crystals. The ensemble of NMR conformers was determined with protein samples labeled with 15N and 13C + 15N. The X-ray structure and NMR ensemble were closely similar with rmsd 1.4 A for residues 8-93. At1g77540 was found to adopt a fold similar to that of GCN5-related N-acetyltransferases. Enzymatic activity assays established that At1g77540 possesses weak acetyltransferase activity against histones H3 and H4. Chemical shift perturbations observed in 15N-HSQC spectra upon the addition of CoA indicated that the cofactor binds and identified its binding site. The molecular details of this interaction were further elucidated by solving the X ray structure of the At1g77540-CoA complex. This work establishes that the domain family COG2388 represents a novel class of acetyltransferase and provides insight into possible mechanistic roles of the conserved Cys76 and His41 residues of this family. PMID- 17128972 TI - Structural analysis and assembly of the HIV-1 Gp41 amino-terminal fusion peptide and the pretransmembrane amphipathic-at-interface sequence. AB - The amino-terminal region within the HIV-1 gp41 aromatic-rich pretransmembrane domain is an amphipathic-at-interface sequence (AIS). AIS is highly conserved between different viral strains and isolates and recognized by the broadly neutralizing 2F5 antibody. The atomic structure of the native Fab2F5-bound AIS appears to involve a nonhelical extended region and a beta-turn structure. We previously described how an immunogenic complex forms, based on the stereospecific interactions between AIS and the gp41 amino-terminal fusion peptide (FP). Here, we have analyzed the structure generated by these interactions using synthetic hybrids containing AIS and FP sequences connected through flexible tethers. The monoclonal 2F5 antibody recognized FP-AIS hybrid sequences with an apparently higher affinity than the linear AIS. Indeed, these hybrids exhibited a weaker capacity to destabilize membranes than FP alone. A combined structural analysis, including circular dichroism, infrared spectroscopy, and two-dimensional infrared correlation spectroscopy, revealed the existence of specific conformations in FP-AIS hybrids, predominantly involving beta-turns. Thermal denaturation studies indicated that FP stabilizes the nonhelical folded AIS structure. We propose that the assembly of the FP-AIS complex may act as a kinetic trap in halting the capacity of FP to promote fusion. PMID- 17128973 TI - Identification of functional domains of Clostridium septicum alpha toxin. AB - Alpha toxin (AT) is the major virulence factor of Clostridium septicum that is a proteolytically activated pore-forming toxin that belongs to the aerolysin-like family of toxins. AT is predicted to be a three-domain molecule on the basis of its functional and sequence similarity with aerolysin, for which the crystal structure has been determined. In this study, we have substituted the entire primary structure of AT with alanine or cysteine to identify those amino acids that comprise functional domains involved in receptor binding, oligomerization, and pore formation. These studies revealed that receptor binding is restricted to domain 1 of the AT structure, whereas domains 1 and 3 are involved in oligomerization. These studies also revealed the presence of a putative functional region of AT proximal to the receptor-binding domain but distal from the pore-forming domain that is proposed to regulate the insertion of the transmembrane beta-hairpin of the prepore oligomer. PMID- 17128974 TI - Identification of the critical residues of bradykinin receptor B1 for interaction with the kinins guided by site-directed mutagenesis and molecular modeling. AB - We report the critical residues for the interaction of the kinins with human bradykinin receptor 1 (B1) using site-directed mutagenesis in conjunction with molecular modeling of the binding modes of the kinins in the homology model of the B1 receptor. Mutation of Lys118 in transmembrane (TM) helix 3, Ala270 in TM6, and Leu294 in TM7 causes a significant decrease in the affinity for the peptide agonists des-Arg10kallidin (KD) and des-Arg9BK but not the peptide antagonist des Arg10Leu9KD. In contrast, mutations in TM2, TM3, TM6, and TM7 cause a significant decrease in the affinity for both the peptide agonists and the antagonist. These data indicate that the B1 bradykinin binding pocket for agonists and antagonists is similar, but the manners in which they interact with the receptor do not completely overlap. Therefore, there is a potential to influence the receptor's ligand selectivity. PMID- 17128975 TI - Analysis of the Cob(II)alamin-5'-deoxy-3',4'-anhydroadenosyl radical triplet spin system in the active site of diol dehydrase. AB - A triplet spin system (S=1) is detected by low-temperature electron paramagnetic resonance (EPR) spectroscopy in samples of diol dehydrase and the functional adenosylcobalamin (AdoCbl) analogue 5'-deoxy-3',4'-anhydroadenosylcobalamin (anAdoCbl). Different spectra are observed in the presence and absence of the substrate (R,S)-1,2-propanediol. In both cases, the spectra include a prominent half-field transition (DeltaM(S) = 2) that is a hallmark of strongly coupled triplet spin systems. The appearance of 59Co hyperfine splitting in the EPR signals and the positions (g values) of the signals in the spectra show that half of the triplet spin is contributed by the low-spin Co2+ of cob(II)alamin. Line width effects from isotopic labeling (13C and 2H) in the 5'-deoxy-3',4' anhydroribosyl ring demonstrate that the other half of the spin triplet is from an allylic 5'-deoxy-3',4'-anhydroadenosyl (anhydroadenosyl) radical. The zero field splitting (ZFS) tensors describing the magnetic dipole-dipole interactions of the component spins of the triplets have rhombic symmetry because of electron spin delocalization within the organic radical component and the proximity of the radical to the low-spin Co2+. The dipole-dipole interaction was modeled as a summation of point-dipole interactions involving the spin-bearing orbitals of the anhydroadenosyl radical and cob(II)alamin. Geometries which are consistent with the ZFS tensors in the presence and absence of the substrate position the 5' carbon of the anhydroadenosyl radical 3.5 and 4.1 A from Co2+, respectively. Homolytic cleavage of the cobalt-carbon bond of the analogue in the absence of the substrate indicates that, in diol dehydrase, binding of the coenzyme to the protein weakens the bond prior to binding of the substrate. PMID- 17128976 TI - Characterization of four covalently-linked yeast cytochrome c/cytochrome c peroxidase complexes: Evidence for electrostatic interaction between bound cytochrome c molecules. AB - Four covalent complexes between recombinant yeast cytochrome c and cytochrome c peroxidase (rCcP) were synthesized via disulfide bond formation using specifically designed protein mutants (Papa, H. S., and Poulos, T. L. (1995) Biochemistry 34, 6573-6580). One of the complexes, designated V5C/K79C, has cysteine residues replacing valine-5 in rCcP and lysine-79 in cytochrome c with disulfide bond formation between these residues linking the two proteins. The V5C/K79C complex has the covalently bound cytochrome c located on the back-side of cytochrome c peroxidase, approximately 180 degrees from the primary cytochrome c-binding site as defined by the crystallographic structure of the 1:1 noncovalent complex (Pelletier, H., and Kraut J. (1992) Science 258, 1748-1755). Three other complexes have the covalently bound cytochrome c located approximately 90 degrees from the primary binding site and are designated K12C/K79C, N78C/K79C, and K264C/K79C, respectively. Steady-state kinetic studies were used to investigate the catalytic properties of the covalent complexes at both 10 and 100 mM ionic strength at pH 7.5. All four covalent complexes have catalytic activities similar to those of rCcP (within a factor of 2). A comprehensive study of the ionic strength dependence of the steady-state kinetic properties of the V5C/K79C complex provides evidence for significant electrostatic repulsion between the two cytochromes bound in the 2:1 complex at low ionic strength and shows that the electrostatic repulsion decreases as the ionic strength of the buffer increases. PMID- 17128977 TI - Specific effects of potassium ion binding on wild-type and L358P cytochrome P450cam. AB - The camphor monoxygenase cytochrome P450cam (CYP101) requires potassium ion (K+) to drive formation of the characteristic high-spin state of the heme Fe+3 upon substrate binding. Amide 1H, 15N correlations in perdeuterated [U-15N] CYP101 were monitored as a function of K+ concentration by 2D-TROSY-HSQC in both camphor bound oxidized (CYP-S) and camphor- and CO-bound reduced CYP101 (CYP-S-CO). In both forms, K+-induced spectral perturbations are detected in the vicinity of the K+ binding site proposed from crystallographic structures, but are larger and more widespread structurally in CYP-S than in CYP-S-CO. In CYP-S-CO, K+-induced perturbations occur primarily near the proposed K+ binding site in the B-B' loop and B' helix, which are also perturbed by binding of effector, putidaredoxin (Pdx). The spectral effects of K+ binding in CYP-S-CO oppose those observed upon Pdxr titration. However, Pdxr titration of CYP-S-CO in the absence of K+ results in multiple conformations. The spin-state equilibrium in the L358P mutant of CYP101 is more sensitive to K+ concentration than WT CYP101, consistent with a hypothesis that L358P preferentially populates conformations enforced by Pdx binding in WT CYP101. Thallium(I), a K+ mimic, minimizes the effects of Pdx titration on the NMR spectrum of CYP-S-CO, but is competent to replace K+ in driving the formation of high-spin CYP-S. These observations suggest that the role of K+ is to stabilize conformers of CYP-S that drive the spin-state change prior to the first electron transfer, and that K+ stabilizes the CYP-S-CO conformer that interacts with Pdx. However, upon binding of Pdx, further conformational changes occur that disfavor K+ binding. PMID- 17128978 TI - Higher order structure contributes to specific differences in redox potential and electron transfer efficiency of root and leaf ferredoxins. AB - Plant type ferredoxin (Fd) is a small [2Fe-2S] cluster containing electron transfer protein with a highly negative redox potential. Higher plants contain different iso-protein types of Fd in roots and leaves, reflecting the difference in redox cascades between these two tissues. We have combined subdomains of leaf and root Fds in recombinant chimeras, to examine structural effects and the relationship between groups of residues on redox potential, electron transfer, and protein-protein interactions. All chimeras had redox potentials that were intermediate to the wild type leaf and root Fds. Surprisingly, the largest differences resulted from exchange of the N-terminus, the region farthest from the redox center. Homology modeling and energy minimization calculations suggest that the N-terminal chimeras may indirectly influence redox potentials by structurally perturbing the active site. Measurements of electron transport and protein interaction indicate that synergistic interaction between the C- and N terminal of root Fd bestows a specific high affinity for accepting electrons in the root type electron cascade, and that there is discrimination against photosynthetic electron donation to root Fd based on the C-terminus of the molecule. Taken together, the experimental and computational studies support a model in which higher order structure contributes to iso-protein specific interaction and electron-transfer properties. PMID- 17128979 TI - Characterization of an HMG-CoA reductase from Listeria monocytogenes that exhibits dual coenzyme specificity. AB - HMG-CoA reductase (HMGR) is an enzyme critical for cellular cholesterol synthesis in mammals and isoprenoid synthesis in certain eubacteria, catalyzing the NAD(P)H dependent reduction of HMG-CoA to mevalonate. We have isolated the gene encoding HMG-CoA reductase from Listeria monocytogenes and expressed the recombinant 6x His-tagged form in Escherichia coli. Using NAD(P)(H), the enzyme catalyzes HMG CoA reduction approximately 200-fold more efficiently than mevalonate oxidation in vitro. The purified enzyme exhibits dual coenzyme specificity, utilizing both NAD(H) and NADP(H) in catalysis; however, catalytic efficiency using NADP(H) is approximately 200 times greater than when using NAD(H). The statins mevinolin and mevastatin are weak inhibitors of L. monocytogenes HMG-CoA reductase, requiring micromolar concentrations for inhibition. Three-dimensional modeling reveals that the overall structure of L. monocytogenes HMG-CoA reductase is likely similar to the known structure of the class II enzyme from Pseudomonas mevalonii. It appears that the enzyme has catalytic amino acids in analogous positions that likely play similar roles and also has a flap domain that brings a catalytic histidine into the active site. However, in L. monocytogenes HMG-CoA reductase histidine 143 and methionine 186 are present in the putative NAD(P)(H)-selective site, possibly interacting with the 2' phosphate of NADP(H) or 2' hydroxyl of NAD(H) and providing the active site architecture necessary for dual coenzyme specificity. PMID- 17128980 TI - A structural limitation on enzyme activity: the case of HMG-CoA synthase. AB - Recent structural studies of the HMG-CoA synthase members of the thiolase superfamily have shown that the catalytic loop containing the nucleophilic cysteine follows the phi and psi angle pattern of a II' beta turn. However, the i + 1 residue is conserved as an alanine, which is quite unusual in this position as it must adopt a strained positive phi angle to accommodate the geometry of the turn. To assess the effect of the conserved strain in the catalytic loop, alanine 110 of Enterococcus faecalis 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase was mutated to a glycine. Subsequent enzymatic studies showed that the overall reaction rate of the enzyme was increased 140-fold. An X-ray crystallographic study of the Ala110Gly mutant enzyme demonstrated unanticipated adjustments in the active site that resulted in additional stabilization of all three steps of the reaction pathway. The rates of acetylation and hydrolysis of the mutant enzyme increased because the amide nitrogen of Ser308 shifts 0.4 A toward the catalytic cysteine residue. This motion positions the nitrogen to better stabilize the intermediate negative charge that develops on the carbonyl oxygen of the acetyl group during both the formation of the acyl-enzyme intermediate and its hydrolysis. In addition, the hydroxyl of Ser308 rotates 120 degrees to a position where it is able to stabilize the carbanion intermediate formed by the methyl group of the acetyl-S-enzyme during its condensation with acetoacetyl-CoA. PMID- 17128981 TI - Roles of Tyr13 and Phe219 in the unique substrate specificity of pepsin B. AB - Pepsin B is known to be distributed throughout mammalia, including carnivores. In this study, the proteolytic specificity of canine pepsin B was clarified with 2 protein substrates and 37 synthetic octapeptides and compared with that of human pepsin A. Pepsin B efficiently hydrolyzed gelatin but very poorly hydrolized hemoglobin. It was active against only a group of octapeptides with Gly at P2, such as KPAGF/LRL and KPEGF/LRL (arrows indicate cleavage sites). In contrast, pepsin A hydrolyzed hemoglobin but not gelatin and showed high activity against various types of octapeptides, such as KPAEF/FRL and KPAEF/LRL. The specificity of pepsin B is unique among pepsins, and thus, the enzyme provides a suitable model for analyzing the structure and function of pepsins and related aspartic proteinases. Because Tyr13 and Phe219 in/around the S2 subsites (Glu/Ala13 and Ser219 are common in most pepsins) appeared to be involved in the specificity of pepsin B, site-directed mutagenesis was undertaken to replace large aromatic residues with small residues and vice versa. The Tyr13Ala/Phe219Ser double mutant of pepsin B was found to demonstrate broad activity against hemoglobin and various octapeptides, whereas the reverse mutant of pepsin A had significantly decreased activity. According to molecular modeling of pepsin B, Tyr13 OH narrows the substrate-binding space and a peptide with Gly at P2 might be preferentially accommodated because of its high flexibility. The hydroxyl can also make a hydrogen bond with nitrogen of a P3 residue and fix the substrate main chain to the active site, thus restricting the flexibility of the main chain and strengthening preferential accommodation of Gly at P2. The phenyl moiety of Phe219 is bulky and narrows the S2 substrate space, which also leads to a preference for Gly at P2, while lowering the catalytic activity against other peptide types without making a hydrogen-bonding network in the active site. PMID- 17128982 TI - Structure and biosynthesis of free lipid A molecules that replace lipopolysaccharide in Francisella tularensis subsp. novicida. AB - Francisella tularensis subsp. novicida U112 phospholipids, extracted without hydrolysis, consist mainly of phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, and two lipid A species, designated A1 and A2. These lipid A species, present in a ratio of 7:1, comprise 15% of the total phospholipids, as judged by 32Pi labeling. Although lipopolysaccharide is detectable in F. tularensis subsp. novicida U112, less than 5% of the total lipid A is covalently linked to it. A1 and A2 were analyzed by electrospray ionization and matrix assisted laser desorption ionization mass spectrometry, gas chromatography/mass spectrometry, and NMR spectroscopy. Both compounds are disaccharides of glucosamine, acylated with primary 3-hydroxystearoyl chains at positions 2, 3, and 2' and a secondary palmitoyl residue at position 2'. Minor isobaric species and some lipid A molecules containing a 3-hydroxypalmitoyl chain in place of 3 hydroxystearate are also present. The 4'- and 3'-positions of A1 and A2 are not derivatized, and 3-deoxy-d-manno-octulosonic acid (Kdo) is not detectable. The 1 phosphate groups of both A1 and A2 are modified with an alpha-linked galactosamine residue, as shown by NMR spectroscopy and gas chromatography/mass spectrometry. An alpha-linked glucose moiety is attached to the 6'-position of A2. The lipid A released by mild acid hydrolysis of F. tularensis subsp. novicida lipopolysaccharide consists solely of component A1. F. tularensis subsp. novicida mutants lacking the arnT gene do not contain a galactosamine residue on their lipid A. Formation of free lipid A in F. tularensis subsp. novicida might be initiated by an unusual Kdo hydrolase present in the membranes of this organism. PMID- 17128983 TI - Rhodopseudomonas palustris CGA009 has two functional ppsR genes, each of which encodes a repressor of photosynthesis gene expression. AB - The PpsR protein is a regulator of redox-dependent photosystem development in purple phototrophic bacteria. In contrast to most species, Rhodopseudomonas palustris contains two ppsR genes. We show that the inactivation of each of the R. palustris strain CGA009 ppsR genes results in an elevated level of formation of the photosystem under dark aerobic conditions. Absorption spectra of the two PpsR mutants revealed qualitative and quantitative differences in light harvesting peak amplitude increases. A sequence difference in the helix-turn helix DNA binding motif of PpsR2 (Arg 439 to Cys) between R. palustris strains CEA001 and CGA009 is shown to be a natural polymorphism that does not inactivate the repressor activity of the protein. To evaluate which photosynthesis genes are regulated by the two PpsR proteins, transcriptome profiles of the CGA009 and PpsR mutant strains were analyzed in microarray experiments. Transcription of most but not all photosystem genes was derepressed in the mutant strains to levels consistent with the in vivo absorption spectra, mathematical analyses of peak shapes and amplitudes, reaction center protein levels, and real-time PCR of selected mRNAs. Closely spaced PpsR binding motif repeats were identified 5' of genes that were derepressed in the transcriptome analysis of PpsR mutants. This work shows that both the PpsR1 and PpsR2 proteins from R. palustris strain CGA009 function as oxygen-responsive transcriptional repressors. PMID- 17128984 TI - Competitive binding of protein kinase Calpha to membranes and Rho GTPases. AB - Previously, we have shown that protein kinase C (PKC) forms a direct high affinity, isozyme-specific and membrane lipid-independent interaction with Rho GTPases [Slater, S. J., Seiz, J. L., Stagliano, B. A., and Stubbs, C. D. (2001) Biochemistry 40, 4437-4445]. Since the cellular activation of PKCalpha involves an initial translocation from cytosolic to membrane compartments, the present study investigates the interdependence between the direct protein-protein interaction of PKCalpha with the Rho GTPase, Cdc42, and the protein-lipid interactions of PKCalpha with membranes. It was hypothesized that the interaction of PKCalpha with membrane-bound Cdc42 would contribute to the overall membrane binding affinity of the kinase by providing an additional anchor. However, it was found that the incorporation of isoprenylated Cdc42 into membranes resulted in an apparent decrease in the membrane-binding affinity of PKCalpha, whereas the association of PKCbetaI, PKCdelta, PKCepsilon, and PKCzeta was each unaffected. The presence of membrane-bound Cdc42 resulted in a rightward shift in both the PS and Ca2+-concentration response curves for PKCalpha membrane association and for the ensuing activation, whereas the maximal levels of binding and activation attained at saturating PS and Ca2+ concentrations were in each case unaffected. Overall, these findings suggest that PKCalpha undergoes a isozyme-specific interaction with membrane-bound Cdc42 to form a PKCalpha-Cdc42 complex, which possesses a membrane-binding affinity that is reduced relative to that of the individual components due to competition between Cdc42 and PS/Ca2+ for binding to PKCalpha. Consistent with this, it was found that the interaction of PKCalpha with membrane-bound Cdc42 was accompanied by the physical dissociation of the PKCalpha-Cdc42 complex from membranes. Thus, the study provides a novel mechanism by which the membrane association and activation of PKCalpha and Cdc42 may be regulated by competing protein-protein and protein-lipid interactions. PMID- 17128985 TI - The temperature dependence of salt-protein association is sequence specific. AB - Molecular dynamics (MD) simulations are used to probe the origin of the unexpected temperature dependence of salt accumulation in the C-terminal region of the protein human lymphotactin. As in previous MD simulations, sodium ions accumulate in an enhanced manner near the C-terminal helix at the lower temperature, while the temperature dependence of chloride accumulation is much weaker and slightly positive. In a designed mutant in which all positively charged residues in the C-terminal helix are replaced with neutral polar groups (Ser), the unexpected temperature dependence of the sodium ions is no longer observed. Therefore, these simulations convincingly verified the previous hypothesis that the temperature dependence of ion-protein association is sensitive to the local sequence. This is explained qualitatively in terms of the entropy of association between charged species in solution. These findings have general implications for the interpretation of thermodynamic quantities associated with binding events where ion release is important, such as protein DNA interactions. PMID- 17128986 TI - Molecular basis for differential nucleotide binding of the nucleotide-binding domain of ABC-transporter CvaB. AB - The cytoplasmic membrane protein CvaB, involved in colicin V secretion in Escherichia coli, belongs to the ABC-transporter family in which ATP hydrolysis is typically the driving force for substrate transport. However, our previous studies indicated that the nucleotide-binding domain of CvaB could also bind and hydrolyze GTP and, indeed, highly preferred GTP over ATP at low temperatures. In this study, we have examined the molecular basis of this preference. Sequence alignment and homology modeling of the CvaB nucleotide-binding domain predicted that the aromatic stacking region of CvaB (Y501DSQ loop) had a role in the differential binding of nucleotides, and Ser503 and Gln504 provided potential hydrogen bonds to GTP but not to ATP. Site-directed mutagenesis of the Y501DSQ loop, mutations S503A, Q504L, and double mutation S503A/Q504L, was made to test the predicted hydrogen bonds with GTP. The double mutation S503A/Q504L increased the affinity for ATP by 6-fold, whereas the affinity for GTP was reduced slightly: the ATP/GTP-binding ratio increased about 10-fold. The temperature effect assays on nucleotide binding and hydrolysis further indicated that the double mutant protein had largely eliminated the difference for substrates ATP and GTP, and behaved more similarly to the NBD of typical ABC-transporter HlyB. Therefore, we conclude that Ser503 and Gln504 in aromatic stacking region of CvaB block the ATP binding and are important for the GTP-binding preference. PMID- 17128987 TI - Redox regulation of RhoA. AB - We have previously shown that redox agents including superoxide anion radical and nitrogen dioxide can react with GXXXXGK(S/T)C motif-containing GTPases (i.e., Rac1, Cdc42, and RhoA) to stimulate guanine nucleotide release. We now show that the reaction of RhoA with redox agents leads to different functional consequences from that of Rac1 and Cdc42 due to the presence of an additional cysteine (GXXXCGK(S/T)C) in the RhoA redox-active motif. While reaction of redox agents with RhoA stimulates guanine nucleotide dissociation, RhoA is subsequently inactivated through formation of an intramolecular disulfide that prevents guanine nucleotide binding thereby causing RhoA inactivation. Thus, redox agents may function to downregulate RhoA activity under conditions that stimulate Rac1 and Cdc42 activity. The opposing functions of these GTPases may be due in part to their differential redox regulation. In addition, the results presented herein suggest that the platinated-chemotherapeutic agent, cisplatin, which is known for targeting nucleic acids, reacts with RhoA to produce a RhoA thiol-cisplatin-thiol adduct, leading to inactivation of RhoA. Similarly, certain arsenic complexes (i.e., arsenate and arsenic trioxide) may inactivate RhoA by bridging the cysteine residues in the GXXXCGK(S/T)C motif. Thus, in addition to redox agents, platinated-chemotherapeutic agents and arsenic complexes may modulate the activity of GTPases containing the GXXXCGK(S/T)C motif (i.e., RhoA and RhoB). PMID- 17128988 TI - Polar destabilization of DNA duplexes with single-stranded overhangs by the Deinococcus radiodurans SSB protein. AB - The Deinococcus radiodurans SSB protein has an occluded site size of 50 +/- 2 nucleotides on ssDNA but can form a stable complex with a 26-30-nucleotide oligodeoxynucleotide using a subset of its four ssDNA binding domains. Quantitative estimates of D. radiodurans SSB protein in the D. radiodurans cell indicate approximately 2500-3000 dimers/cell, independent of the level of irradiation. At biologically relevant concentrations, when bound at single-strand double-strand DNA junctions in vitro, D. radiodurans SSB protein has a limited capacity to displace the shorter strand of the duplex, permitting it to bind to single-strand extensions shorter than 26-30 nucleotides. The capacity to displace the shorter strand of the duplex shows a pronounced bias for extensions with a free 3' end. The Escherichia coli SSB protein has a similar but somewhat less robust capacity to displace a DNA strand annealed adjacent to a single-strand extension. These activities are likely to be relevant to the action of bacterial SSB proteins in double-strand break repair, acting at the frayed ends created by ionizing radiation. PMID- 17128989 TI - N- and C-terminal hydrophobic patches are involved in fibrillation of glucagon. AB - Recent work suggests that the molecular structure of amyloid-like fibrils is determined by environmental conditions as well as amino acid sequence. To probe the involvement of side chains in fibrillation of the 29-residue hormone glucagon, we have measured fibrillation kinetics of 15 alanine mutants. At acidic pH, all of the mutants are able to form fibrils. However, substitution of hydrophobic residues in the N- and C-termini (in particular Phe6, Tyr10, Val23, and Met27) decelerates fibrillation dramatically. This indicates that the hydrophobicity and/or high beta-sheet propensity of these residues may be important for fibrillation. In contrast, substitution of Leu14 increases fibrillation propensity compared to that of the wild type. Nevertheless, despite identical fibrillation conditions, the thioflavin T and tryptophan fluorescence spectra of fibrils formed by mutants Tyr13, Leu14, and Asp15 are significantly different from those of other mutants, indicating that substitution of these residues may influence not only the fibrillation kinetics and fibril stability but also the preferred final structure of the fibrils that is formed, in line with the general structural polymorphism of glucagon fibrils. In contrast, under alkaline conditions, only a handful of the alanine mutants are capable of forming fibrils, suggesting that more side chains are involved in stabilizing interactions here. In addition, fibrils formed by wild-type glucagon at alkaline pH appear very stable, compared to fibrils formed at acidic pH. This suggests that the distribution of charges determines the number of different fibrillated states available to a peptide, since these can block formation of metastable fibrillated states. PMID- 17128990 TI - (+/-)-(1S,2R,5S)-5-Amino-2-fluorocyclohex-3-enecarboxylic acid. A potent GABA aminotransferase inactivator that irreversibly inhibits via an elimination aromatization pathway. AB - Inhibition of gamma-aminobutyric acid aminotransferase (GABA-AT) increases the concentration of GABA, an inhibitory neurotransmitter in human brain, which could have therapeutic applications for a variety of neurological diseases, including epilepsy. On the basis of studies of several previously synthesized conformationally restricted GABA-AT inhibitors, (+/-)-(1S,2R,5S)-5-amino-2 fluorocyclohex-3-enecarboxylic acid (12) was designed as a mechanism-based inactivator. This compound was shown to irreversibly inhibit GABA-AT; substrate protects the enzyme from inactivation. Mechanistic experiments demonstrated the loss of one fluoride ion per active site during inactivation and the formation of N-m-carboxyphenylpyridoxamine 5'-phosphate (26), the same product generated by inactivation of GABA-AT by gabaculine (8). An elimination-aromatization mechanism is proposed to account for these results. PMID- 17128991 TI - Intermediates in assembly by photoactivation after thermally accelerated disassembly of the manganese complex of photosynthetic water oxidation. AB - The Mn4Ca complex bound to photosystem II (PSII) is the active site of photosynthetic water oxidation. Its assembly involves binding and light-driven oxidation of manganese, a process denoted as photoactivation. The disassembly of the Mn complex is a thermally activated process involving distinct intermediates. Starting from intermediate states of the disassembly, which was initiated by a temperature jump to 47 degrees C, we photoactivated PSII membrane particles and monitored the activity recovery by O2 polarography and delayed chlorophyll fluorescence measurements. Oxidation state and structural features of the formed intermediates of the Mn complex were assayed by X-ray absorption spectroscopy at the Mn K-edge. The photoactivation time courses, which exhibit a lag phase characteristic of intermediate formation only when starting with the apo-PSII, suggest that within approximately 5 min of photoactivation of apo-PSII, a binuclear Mn complex is formed. It is proposed that a MnIII2(di-mu-oxo) complex is a key intermediate both in the disassembly and in the assembly reaction paths. PMID- 17128992 TI - The N-terminal domain of Bcl-xL reversibly binds membranes in a pH-dependent manner. AB - Bcl-xL regulates apoptosis by maintaining the integrity of the mitochondrial outer membrane by adopting both soluble and membrane-associated forms. The membrane-associated conformation does not require a conserved, C-terminal transmembrane domain and appears to be inserted into the bilayer of synthetic membranes as assessed by membrane permeabilization and critical surface pressure measurements. Membrane association is reversible and is regulated by the cooperative binding of approximately two protons to the protein. Two acidic residues, Glu153 and Asp156, that lie in a conserved hairpin of Bcl-xLDeltaTM appear to be important in this process on the basis of a 16% increase in the level of membrane association of the double mutant E153Q/D156N. Contrary to that for the wild type, membrane permeabilization for the mutant is not correlated with membrane association. Monolayer surface pressure measurements suggest that this effect is primarily due to less membrane penetration. These results suggest that E153 and D156 are important for the Bcl-xLDeltaTM conformational change and that membrane binding can be distinct from membrane permeabilization. Taken together, these studies support a model in which Bcl-xL activity is controlled by reversible insertion of its N-terminal domain into the mitochondrial outer membrane. Future studies with Bcl-xL mutants such as E153Q/D156N should allow determination of the relative contributions of membrane binding, insertion, and permeabilization to the regulation of apoptosis. PMID- 17128994 TI - Energy-linked binding of Pi is required for continuous steady-state proton translocating ATP hydrolysis catalyzed by F0.F1 ATP synthase. AB - The presence of medium Pi (half-maximal concentration of 20 microM at pH 8.0) was found to be required for the prevention of the rapid decline in the rate of proton-motive force (pmf)-induced ATP hydrolysis by Fo.F1 ATP synthase in coupled vesicles derived from Paracoccus denitrificans. The initial rate of the reaction was independent of Pi. The apparent affinity of Pi for its "ATPase-protecting" site was strongly decreased with partial uncoupling of the vesicles. Pi did not reactivate ATPase when added after complete time-dependent deactivation during the enzyme turnover. Arsenate and sulfate, which was shown to compete with Pi when Fo.F1 catalyzed oxidative phosphorylation, substituted for Pi as the protectors of ATPase against the turnover-dependent deactivation. Under conditions where the enzyme turnover was not permitted (no ATP was present), Pi was not required for the pmf-induced activation of ATPase, whereas the presence of medium Pi (or sulfate) delayed the spontaneous deactivation of the enzyme which was induced by the membrane de-energization. The data are interpreted to suggest that coupled and uncoupled ATP hydrolysis catalyzed by Fo.F1 ATP synthases proceeds via different intermediates. Pi dissociates after ADP if the coupling membrane is energized (no E.ADP intermediate exists). Pi dissociates before ADP during uncoupled ATP hydrolysis, leaving the E.ADP intermediate which is transformed into the inactive ADP(Mg2+)-inhibited form of the enzyme (latent ATPase). PMID- 17128993 TI - Surfactant protein A without the interruption of Gly-X-Y repeats loses a kink of oligomeric structure and exhibits impaired phospholipid liposome aggregation ability. AB - Pulmonary surfactant protein A (SP-A) belongs to the collectin subgroup of the C type lectin superfamily. SP-A oligomerizes as an octadecamer, which forms a flower bouquet-like structure. A collagen-like domain of human SP-A consists of 23 Gly-X-Y repeats with an interruption near the midpoint of this domain. This interruption causes a kink, but its role remains unknown. To define the importance of the kink region of SP-A, two mutated proteins were constructed to disrupt the interruption of Gly-X-Y repeats: SP-ADEL, which lacks the Pro47-Cys48 Pro49-Pro50 sequence at the interruption, and SP-AINS, in which two glycines were introduced to insert Gly-X-Y repeats (Gly-Pro47-Cys48-Gly-Pro49-Pro50). Electron microscopy using rotary shadowing revealed that both mutants form octadecamers that lack a bend in the collagenous domain. Electrophoretic analysis under nondenaturing conditions and gel filtration chromatography demonstrated that SP AINS consisted of a large assembly of oligomers whereas SP-ADEL formed mainly octadecamers. Both SP-ADEL and SP-AINS mutants as well as wild-type SP-A bound to liposomes containing dipalmitoylphosphatidylcholine and galactosylceramide at equivalent levels, but the abilities of the mutants to induce phospholipid liposome aggregation were significantly less developed than that of the wild type. The mutants SP-ADEL and SP-AINS augmented liposome uptake by alveolar type II cells and inhibited secretion of phospholipids from type II cells at a level comparable to that of wild-type SP-A. These results indicate that the interruption of Gly-X-Y repeats in the SP-A molecule is critical for the formation of a flower bouquet-like octadecamer and contributes to SP-A's capacity to aggregate phospholipid liposomes. PMID- 17128996 TI - Topographic, electrochemical, and optical images captured using standing approach mode scanning electrochemical/optical microscopy. AB - We developed a high-resolution scanning electrochemical microscope (SECM) for the characterization of various biological materials. Electrode probes were fabricated by Ti/Pt sputtering followed by parylene C-vapor deposition polymerization on the pulled optical fiber or glass capillary. The effective electrode radius estimated from the cyclic voltammogram of ferrocyanide was found to be 35 nm. The optical aperture size was less than 170 nm, which was confirmed from the cross section of the near-field scanning optical microscope (NSOM) image of the quantum dot (QD) particles with diameters in the range of 10-15 nm. The feedback mechanism controlling the probe-sample distance was improved by vertically moving the probe by 0.1-3 microm to reduce the damage to the samples. This feedback mode, defined as "standing approach (STA) mode" (Yamada, H.; Fukumoto, H.; Yokoyama, T.; Koike, T. Anal. Chem. 2005, 77, 1785-1790), has allowed the simultaneous electrochemical and topographic imaging of the axons and cell body of a single PC12 cell under physiological conditions for the first time. STA-mode feedback imaging functions better than tip-sample regulation by the conventionally available AFM. For example, polystyrene beads (diameter approximately 6 microm) was imaged using the STA-mode SECM, whereas imaging was not possible using a conventional AFM instrument. PMID- 17128997 TI - Scanning electrochemical microscopy determination of organic soluble MPC electron transfer rates. AB - In this paper, we describe a novel method for measuring the forward heterogeneous electron-transfer rate constant (kf) through the thiol monolayer of gold monolayer protected clusters (MPCs) in solution using scanning electrochemical microscopy (SECM). Applying the equations for mixed mass-transfer and electron transfer processes, we develop a new formula using only the diffusion coefficient and the tip radius and use it as part of a new method for evaluating SECM approach curves. This method is applied to determine the electron-transfer rates from a series of SECM approach curves for monodisperse hexanethiol MPCs and for polydisperse hexanethiol, octanethiol, decanethiol, dodecanethiol, and 2 phenyethylthiol gold MPCs. Our results show that as the alkanethiol length increases the rate of electron transfer decreases in a manner consistent with the previously proposed tunneling mechanism for the electron transfer in MPCs. However, the effective tunneling coefficient, Beta, is found to be only 0.41 A-1 for alkanethiol passivated MPCs compared to typical values of 1.1 A-1 for alkanethiols as self-assembled monolayers on two-dimensional gold substrates. Similar SECM approach curve results for Pt and Au MPCs indicate that the electron transfer rate is dependent mostly on the composition of the thiol layer and not on differences in the core metal. PMID- 17128998 TI - In situ EC-STM studies of MPS, SPS, and chloride on Cu100: structural studies of accelerators for dual damascene electrodeposition. AB - Electrochemical, differential capacitance, and in situ electrochemical scanning tunneling microscopy (EC-STM) methods are used to examine the interaction of bis(3-sulfopropyl)-disulfide (SPS) and mercaptopropylsulfonic acid (MPS) with Cu(100) surfaces both in the absence and presence of chloride. Both electrochemical and differential capacitance results are weakly perturbed by the addition of either MPS or SPS in the potential region between -0.2 and -0.5 V versus Ag/AgCl relative to the additive-free case. EC-STM images obtained from solutions of MPS alone exhibit a c(2 x 2) adlattice whereas those from SPS alone yield only the (1 x 1) structure. In the presence of Cl-, both adsorbates evince only a c(2 x 2) adlattice on the Cu(100) surface. The desorption potential of these structure is identical to that found with Cl- alone. These results show that neither MPS nor SPS adsorbs strongly on Cu(100) in the presence of Cl-. PMID- 17128999 TI - Scanning electrochemical mapping of spatially localized electrochemical reactions induced by surface potential gradients. AB - The influence of a surface potential gradient on the location and extent of electrochemical reactions was examined using a scanning electrochemical microscope. A linear potential gradient was imposed on the surface of a platinum coated indium tin oxide electrode by applying two different potential values at the edges of the electrode. The applied potentials were used to control the location and extent of several electrochemical reactions, including the oxidation of Ru(NH3)6(2+), the oxidation of H2, and the oxidation of H2 in the presence of adsorbed CO. Scanning electrochemical mapping of these reactions was achieved by probing the feedback current associated with the oxidation products. The oxidation of Ru(NH3)6(2+) occurred at locations where the applied potential was positive of the formal potential of the Ru(NH3)6(2+/3+) redox couple. The position of this reaction on the surface could be spatially translated by manipulating the terminal potentials. The rate of hydrogen oxidation on the platinum-coated electrode varied spatially in the presence of a potential gradient and correlated with the nature of the electrode surface. High oxidation rates occurred at low potentials, with decreasing rates observed as the potential increased to values where platinum oxides formed. The extent of oxide formation versus position was confirmed with in-situ ellipsometry mapping. In the presence of adsorbed carbon monoxide, a potential gradient created a localized region of high activity for hydrogen oxidation at potentials between where carbon monoxide was adsorbed and platinum oxides formed. The position of this localized region of activity could be readily translated along the surface by changing the terminal potential values. The ability to manipulate electrochemical reactions spatially on a surface has potential application in microscale analytical devices as well as in the discovery and analysis of electrocatalytic systems. PMID- 17129000 TI - Tellurium adatoms as an in-situ surface probe of (111) two-dimensional domains at platinum surfaces. AB - Irreversibly adsorbed tellurium has been studied as a probe to quantify ordered domains in platinum electrodes. The surface redox process of adsorbed tellurium on the Pt(111) electrode and Pt(111) stepped surfaces takes place around 0.85 V in a well-defined peak. The behavior of this redox process on the Pt(111) vicinal surfaces indicates that the tellurium atoms involved in the redox process are only those deposited on the (111) terrace sites. Moreover, the corresponding charge density is proportional to the number of sites on (111) ordered domains (terraces) that are, at least, three atoms wide. Hence, this charge density can be used to measure the number of (111) terrace sites on any given platinum sample. Structural information about tellurium adsorption is obtained from atomic resolution STM images for the Pt(111) and Pt(10, 10, 9) electrodes. A rectangular structure (2 x radical 3) and a compact hexagonal structure (11 x 8) were identified. However, the redox peak for adsorbed tellurium on (100) domains at 1.03 V overlaps with peaks arising from steps and (110) sites. Therefore, it cannot be used without problems for the determination of (100) sites on a platinum sample. On the (100) terraces, the surface structure of the adsorbed tellurium is c(2 x 2), as revealed by STM. Finally, tellurium irreversible adsorption has been used to estimate the number of (111) ordered domains terrace sites on different polycrystalline platinum samples, and the results are compared to those obtained with bismuth irreversible adsorption. PMID- 17129001 TI - Polymer films on electrodes: investigation of ion transport at poly(3,4 ethylenedioxythiophene) films by scanning electrochemical microscopy. AB - Electropolymerization, morphology characterization, and ion transport of poly(3,4 ethylenedioxythiophene) (PEDOT) films doped with different counterions (chloride, ferrocyanide (FCN), and poly(p-styrenesulfonate) (PSS-)) on a platinum electrode were investigated using scanning electrochemical microscopy (SECM) during both potential step (chronocoulometric) and cyclic voltammetric scans. An ultramicroelectrode (UME) tip was positioned close to the surface of a PEDOT modified substrate electrode, and the responses of both electrodes to a substrate potential step or linear sweep were monitored simultaneously. Chloride or ferrocyanide (FCN) ejection during PEDOT reduction was shown to be a function of the reduction potential. The nature of the cation in the bulk solution was not found to be important in the kinetics of ion transport in PEDOT+/FCN- films. Direct evidence for the incorporation of cations of Ru(NH3)6(3+/2+) in a PEDOT film during its reduction was also obtained by SECM measurements. The adsorption of Ru(NH3)6(3+) in fully oxidized PEDOT+/PSS- films was observed and attributed to ion exchange between the Na+ co-ion of PSS- and Ru(NH3)6(3+) in the bulk solution. PMID- 17129002 TI - Conical tungsten tips as substrates for the preparation of ultramicroelectrodes. AB - Here we describe a simple method to prepare voltammetric microelectrodes using tungsten wires as a substrate. Tungsten wires have a high tensile modulus and enable the fabrication of electrodes that have small dimensions overall while retaining rigidity. In this work, 125 microm tungsten wires with a conical tip were employed. For the preparation of gold or platinum ultramicroelectrodes, commercial tungsten microelectrodes, completely insulated except at the tip, were used as substrates. Following removal of oxides from the exposed tungsten, platinum or gold was electroplated, yielding surfaces with an electroactive area of between 1 x 10-6 and 2 x 10-6 cm2. Carbon surfaces on the etched tip of tungsten microwires were prepared by coating with photoresist followed by pyrolysis. The entire electrode was then insulated with Epoxylite except the tip, yielding an exposed carbon surface with an area of around 4 x 10-6 to 6 x 10-6 cm2. All three types of ultramicroelectrodes fabricated on the tungsten wire had similar electrochemical behavior to electrodes fabricated from wires or fibers insulated with glass tubes. PMID- 17129003 TI - Surface-enhanced hyper-Raman scattering (SEHRS) on Ag film over Nanosphere (FON) electrodes: surface symmetry of centrosymmetric adsorbates. AB - Electrochemical surface-enhanced hyper-Raman scattering (SEHRS) and surface enhanced Raman scattering (SERS) of centrosymmetric molecules on Ag film over nanosphere (AgFON) electrodes are presented. The SEHR spectra of trans-1,2-bis(4 pyridyl)ethylene (BPE) at different potentials (vs Ag/AgCl) are presented for the first time, and a reversible potential tuning of the SEHR spectra of BPE is demonstrated. The SEHRS and SERS techniques were used to determine to what extent either site symmetry reduction or field gradient effects dictate the origin of the observed vibrational spectra. It is found that the SEHR and SER spectra for the molecules studied were distinctly different at all frequency regions at a fixed voltage, suggesting that centrosymmetry is largely retained upon adsorption to the AgFON surface and that field gradient effects are negligible. This work also shows that the SEHR spectra clearly depend on potential, whereas the SER spectra are essentially independent of potenial. It is determined that the combination of changes in deltaGads and the presence of coadsorbed counterions are responsible for altering the local symmetry of the adsorbate and only SEHRS has the sensitivity to detect these changes in the surface environment. Thus, SEHRS is a uniquely useful spectroscopic tool that is much more sensitive to the local adsorption environment than is SERS. PMID- 17129004 TI - Layer-by-layer PMIRRAS characterization of DMPC bilayers deposited on a Au111 electrode surface. AB - A combination of Langmuir-Blodgett and Langmuir-Schaefer techniques was employed to deposit 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) bilayers at a gold electrode surface. One leaflet consisted of hydrogen-substituted acyl chains, and the second leaflet was composed of molecules with deuterium-substituted acyl chains. This architecture allowed for layer-by-layer analysis of the structure of the bilayer. Photon polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) was used to determine the conformation and orientation of the acyl chains of DMPC molecules in the individual leaflets as a function of the potential applied to the gold electrode. The bilayer is adsorbed onto the metal surface when the field applied to the membrane does not exceed approximately 108 V/m. When adsorbed, the bottom leaflet is in contact with a hydrophobic metal surface, and the top leaflet is interacting with the aqueous solution. The asymmetry of the environment has an effect on the orientation of the DMPC molecules in each leaflet. The tilt angle of the acyl chains of the DMPC molecules in the bottom leaflet that is in contact with the gold is approximately 10 degrees smaller than that observed for the top leaflet that is exposed to the solution. These studies provide direct evidence that the structure of a phospholipid bilayer deposited at an electrode surface is affected by interaction with the metal. PMID- 17129005 TI - Surface-enhanced Raman spectroscopy using gold-core platinum-shell nanoparticle film electrodes: toward a versatile vibrational strategy for electrochemical interfaces. AB - The aim of this work is to further improve the molecular generality and substrate generality of SERS (i.e., to fully optimize the SERS activity of transition-metal electrodes). We utilized a strategy of borrowing high SERS activity from the Au core based on Au-core Pt-shell (Au@Pt) nanoparticle film electrodes, which can be simply and routinely prepared. The shell thickness from about one to five monolayers of Pt atoms can be well controlled by adjusting the ratio of the number of Au seeds to Pt(IV) ions in the solution. The SERS experimental results of carbon monoxide adsorption indicate that the enhancement factor for the Au@Pt nanoparticle film electrodes is more than 2 orders of magnitude larger than that of electrochemically roughened Pt electrodes. The practical virtues of the present film electrodes for obtaining rich and high-quality vibrational information for diverse adsorbates on transition metals are pointed out and briefly illustrated with systems of CO, hydrogen, and benzene adsorbed on Pt. We believe that the electrochemical applications of SERS will be broadened with this strategy, in particular, for extracting detailed vibrational information for adsorbates at transition-metal electrode interfaces. PMID- 17129006 TI - Measurement of apparent diffusion coefficients within ultrathin nafion Langmuir Schaefer films: comparison of a novel scanning electrochemical microscopy approach with cyclic voltammetry. AB - The use of scanning electrochemical microscopy (SECM) to evaluate the apparent diffusion coefficient, Dapp, of redox-active species in ultrathin Nafion films is described. In this technique, an ultramicroelectrode (UME) tip, positioned close to a film on a macroscopic electrode, is used to oxidize (or reduce) a species in bulk solution, causing the tip-generated oxidant (reductant) to diffuse to the film/solution interface. The oxidation (reduction) of film-confined species regenerates the reductant (oxidant) in solution, leading to feedback to the UME. A numerical model is developed that allows Dapp to be determined. For these studies, ultrathin films of Nafion were prepared using the Langmuir-Schaefer (LS) technique and loaded with an electroactive species, either the ferrocene derivative ferrocenyltrimethylammonium cation, FA+, or tris(2,2' bipyridyl)ruthenium(II), Ru(bpy)32+. The morphology and the thickness of the Nafion LS films (1.5 +/- 0.2 nm per layer deposited) were evaluated using atomic force microscopy (AFM). For comparison with the SECM measurements, cyclic voltammetry (CV) was employed to evaluate the concentration of electroactive species within the Nafion LS films and to determine Dapp. The latter was found to be essentially invariant with film thickness, but the value for Ru(bpy)32+ was 1 order of magnitude larger than for FA+. CV and SECM measurements yield different values of Dapp, and the underlying reasons are discussed. In general, the Dapp values for these films are considerably smaller than for recast Nafion films, which can be attributed to the compactness of Nafion LS films. Nonetheless, the ultrathin nature of the films leads to fast response times, and we thus expect that these modified electrodes could find applications in sensing, electroanalysis, and electrocatalysis. PMID- 17129007 TI - In situ, time-resolved reflectance spectroscopy in the microsecond domain: oxidation of adsorbed carbon monoxide on polycrystalline pt microelectrodes in aqueous solutions. AB - The dynamics of the electrooxidation of adsorbed CO, COads, on polycrystalline Pt microelectrodes has been examined in CO-saturated 0.5 M H2SO4 and 0.5 M HClO4 aqueous solutions, using in situ, time-resolved, normalized differential reflectance spectroscopy lambda = 633 nm). Attention was focused on the unique dependence of COads oxidation on the potential at which the adsorbed full CO monolayer is assembled (i.e., hydrogen adsorption/desorption vs the double-layer region) using both fast linear scan voltammetry and potential step techniques. As evidenced from the data collected, COads oxidation at a fixed potential proceeds at slower rates when the monolayer is formed in the double- layer region compared to when it is formed in the hydrogen adsorption/desorption region. Possible explanations for this effect are discussed. PMID- 17129008 TI - Bridge-bonded formate: active intermediate or spectator species in formic acid oxidation on a Pt film electrode? AB - We present and discuss the results of an in situ IR study on the mechanism and kinetics of formic acid oxidation on a Pt film/Si electrode, performed in an attenuated total reflection (ATR) flow cell configuration under controlled mass transport conditions, which specifically aimed at elucidating the role of the adsorbed bridge-bonded formates in this reaction. Potentiodynamic measurements show a complex interplay between formation and desorption/oxidation of COad and formate species and the total Faradaic current. The notably faster increase of the Faradaic current compared to the coverage of bridge-bonded formate in transient measurements at constant potential, but with different formic acid concentrations, reveals that adsorbed formate decomposition is not rate-limiting in the dominant reaction pathway. If being reactive intermediate at all, the contribution of formate adsorption/decomposition to the reaction current decreases with increasing formic acid concentration, accounting for at most 15% for 0.2 M DCOOH at 0.7 VRHE. The rapid build-up/removal of the formate adlayer and its similarity with acetate or (bi-)sulfate adsorption/desorption indicate that the formate adlayer coverage is dominated by a fast dynamic adsorption desorption equilibrium with the electrolyte, and that formate desorption is much faster than its decomposition. The results corroborate the proposal of a triple pathway reaction mechanism including an indirect pathway, a formate pathway, and a dominant direct pathway, as presented previously (Chen, Y. X.; et al. Angew. Chem. Int. Ed. 2006, 45, 981), in which adsorbed formates act as a site-blocking spectator in the dominant pathway rather than as an active intermediate. PMID- 17129009 TI - Palladium monolayer and palladium alloy electrocatalysts for oxygen reduction. AB - We investigated the oxygen-reduction reaction (ORR) on Pd monolayers on various surfaces and on Pd alloys to obtain a substitute for Pt and to elucidate the origin of their activity. The activity of Pd monolayers supported on Ru(0001), Rh(111), Ir(111), Pt(111), and Au(111) increased in the following order: Pd/Ru(0001) < Pd/Ir(111) < Pd/Rh(111) < Pd/Au(111) < Pd/Pt(111). Their activity was correlated with their d-band centers, which were calculated using density functional theory (DFT). We found a volcano-type dependence of activity on the energy of the d-band center of Pd monolayers, with Pd/Pt(111) at the top of the curve. The activity of the non-Pt Pd2Co/C alloy electrocatalyst nanoparticles that we synthesized was comparable to that of commercial Pt-containing catalysts. The kinetics of the ORR on this electrocatalyst predominantly involves a four electron step reduction with the first electron transfer being the rate determining step. The downshift of the d-band center of the Pd "skin", which constitutes the alloy surface due to the strong surface segregation of Pd at elevated temperatures, determined its high ORR activity. Additionally, it showed very high methanol tolerance, retaining very high catalytic activity for the ORR at high concentrations of methanol. Provided its stability is satisfactory, this catalyst might possibly replace Pt in fuel-cell cathodes, especially those of direct methanol oxidation fuel cells (DMFCs). PMID- 17129010 TI - Cooperativity of copper and molybdenum centers in polyoxometalate-based electrocatalysts: cyclic voltammetry, EQCM, and AFM characterization. AB - Electrochemical behaviors of selected Dawson-type polytungstates including 2 K10[P2W15Mo2O61box] where the symbol [box] designates a vacant site, alpha2 K7[Fe(OH2)P2W15Mo2O61], alpha2-K8[Cu(OH2)P2W15Mo2O61], alpha1- and alpha2 K8[Cu(OH2)P2W17O61], alpha2-K8[Cu(OH2)P2W13Mo4O61], and alpha2 K8[Cu(OH2)P2W12Mo5O61] were investigated by cyclic voltammetry (CV) coupled with the electrochemical quartz microbalance (EQCM), and the results were completed by atomic force microscopy (AFM) observations of the electrodeposited films. The electrocatalytic abilities of these polyoxometalates (POMs) in the reduction of dioxygen, hydrogen peroxide, and NOx were also assessed by CV and EQCM. It turns out that the remarkable electrocatalysis obtained at the reduction potential of Mo centers within alpha2-K8[Cu(OH2)P2W15Mo2O61], but in a domain where Cu2+ is not deposited, benefits from the assistance of the copper center because such catalysis could not be observed in the absence of Cu2+. EQCM confirms that no copper deposition occurs under the experimental conditions used. Analogous behaviors are encountered in the electrocatalytic reduction of nitrite where assistance by the presence of the Cu2+ center induced the observation of catalysis at the potential location of Mo centers. Finally, the reduction of nitrate is triggered by electrodeposited copper but was remarkably favored by the presence of molybdenum atoms within these polyoxometalates (POMs). All of the results converge to indicate a cooperative effect between the Mo and Cu centers within these POMs. The various results suggest that copper deposition from these POMs should give morphologically different surfaces. AFM studies confirm this expectation, and the observed morphologies and sizes of particles were rationalized by taking into account the role of the POM skeleton and its atomic composition in the electrodeposition process. PMID- 17129011 TI - Characterization and theory of electrocatalysts based on scanning electrochemical microscopy screening methods. AB - A strategy for finding new electrocatalysts for the oxygen reduction reaction (ORR) in acidic solutions is outlined and illustrated with results for Pd-Co catalysts. This is based on establishing guidelines for selecting test systems, rapid preparation of arrays, and rapid screening by scanning electrochemical microscopy. Promising candidates are further tested as supported electrocatalysts by larger scale electrochemical methods and in fuel cells, with optimization of the composition and structure. Those that emerge are characterized by a variety of methods, including X-ray diffraction, scanning electron microscopy, and X-ray photoemission spectroscopy. Finally, density functional theory is used for detailed calculations of oxygen adsorption and dissociation on the material and provides better guidelines for further testing. PMID- 17129012 TI - Characterization and analysis of new catalysts for a direct ethanol fuel cell. AB - A direct ethanol fuel cell offers an attractive, fairly high density, energy source, if an electrochemical system can be developed that efficiently carries out the 12-electron oxidation of ethanol to carbon dioxide and water. To that end, new catalyst systems must be developed along with fuel cell operating conditions that encourage the complete oxidation of ethanol, as opposed to the presently available platinum on carbon systems that tend to produce acetaldehyde (two-electron oxidation) or acetic acid (four-electron oxidation) products. It is found that a composite nanoparticulate catalyst containing platinum and tin oxide or platinum indium tin oxide allows the partial conversion of ethanol to its 12 electron oxidation products. Catalysts of this type can be formed using a modified polyol process. Elevation of the operating temperature of a proton exchange membrane fuel cell using the indicated catalysts to 130 degrees C facilitates the production of carbon dioxide and provides an improved current voltage response. PMID- 17129013 TI - Electrochemical surface characterization and O2 reduction kinetics of Se surface modified ru nanoparticle-based RuSe(y)/C catalysts. AB - The electrochemical properties of Se surface-modified Ru/C catalysts (RuSey/C with y = 0 to 1) and their O2 reduction characteristics were determined in model studies under well-defined mass transport conditions, combining quantitative differential electrochemical mass spectrometry and double-disk electrode thin layer flow-cell measurements. Surface characterization of the catalysts including the quantitative evaluation of the active surface area was performed by electrochemical/mass spectrometric (combined H-upd adsorption, preadsorbed CO monolayer oxidation, Cu-upd adsorption/stripping, and RuOx formation) methods. The suitability of these methods for the determination of the active surface area in the high and low Se coverage regime are discussed, and COad stripping is found to be the most relevant method for the present catalysts. The kinetic parameters for the ORR (activity and selectivity) under quasi-steady-state conditions and their variation with Se modification were evaluated in potentiostatic flow-cell measurements. Modification of Ru/C catalyst by Se improves the O2 reduction activity and reduces the tendency for H2O2 formation in the technically relevant potential region of 0.6-0.8 VRHE, but even for the best catalyst compositions a significant ( approximately 0.2 VRHE) overpotential for O2 reduction on the RuSey/C catalysts remains compared to that for the Pt/C catalyst, and we find H2O2 yields of at least 1% at typical cathode operation potentials. Consequences of the relatively high H2O2 yields for membrane/electrode stability in practical applications are discussed. PMID- 17129014 TI - PtRu nanoparticle electrocatalyst with bulk alloy properties prepared through a sonochemical method. AB - Properties of PtRu nanoparticles prepared using high-intensity sonochemistry are reported. Syntheses were carried out in tetrahydrofuran (THF) containing Ru3+ and Pt4+ in a fixed mole ratio of either 1:10 or 1:1. X-ray diffraction measurements confirmed sonocation produces an alloy phase and showed that the composition of the nanometer scale metal particles is close to the mole fraction of Ru3+ and Pt4+ in solution with deviations that tend toward Ru enrichment in the alloy phase. The materials gave responses that are similar in terms of peak potential and current density, referenced to the catalyst active surface area, to those of bulk alloys in voltammetry experiments involving CO stripping and CH3OH electrochemical oxidation in 0.1 M H2SO4. The results show that sonochemical methods have the potential to produce nanometer scale bimetallic electrocatalysts that possess alloy properties. The materials have application in mechanistic studies of fuel cell reactions and as platforms for the development of CO tolerant fuel cell catalyst. PMID- 17129015 TI - Bipolar electrochemical mechanism for the propulsion of catalytic nanomotors in hydrogen peroxide solutions. AB - Bimetallic nanorods are propelled in aqueous solutions by the catalytic decomposition of hydrogen peroxide to oxygen and water. Several mechanisms (interfacial tension gradients, bubble recoil, viscous Brownian ratchet, self electrophoresis) have been proposed for the transduction of chemical to mechanical energy in this system. From Tafel plots of anodic and cathodic hydrogen peroxide reactions at various metal (Au, Pt, Rh, Ni, Ru, and Pd) ultramicroelectrodes, we determine the potential at which the anodic and cathodic reaction rates are equal for each metal. These measurements allow one to predict the direction of motion of all possible bimetallic combinations according to the bipolar electrochemical (or self-electrophoretic) mechanism. These predictions are consistent with the observed direction of motion in all cases studied, providing strong support for the mechanism. We also find that segmented nanorods with one Au end and one poly(pyrrole) end containing catalase, an enzyme that decomposes hydrogen peroxide nonelectrochemically, perform the overall catalytic reaction at a rate similar to that of nanorods containing Au and Pt segments. However, in this case there is no observed axial movement, again supporting the bipolar electrochemical propulsion mechanism for bimetallic nanorods. PMID- 17129016 TI - Methanol dehydrogenation and oxidation on Pt(111) in alkaline solutions. AB - Adsorption, dehydrogenation, and oxidation of methanol on Pt(111) in alkaline solutions has been examined from a fundamental mechanistic perspective, focusing on the role of adsorbate-adsorbate interactions and the effect of defects on reactivity. CO has been confirmed as the main poisoning species, affecting the rate of methanol dehydrogenation primarily through repulsive interactions with methanol dehydrogenation intermediates. At direct methanol fuel cell (DMFC) relevant potentials, methanol oxidation occurs almost entirely through a CO intermediate, and the rate of CO oxidation is the main limiting factor in methanol oxidation. Small Pt island defects greatly enhance CO oxidation, though they are effective only when the CO coverage is 0.20 ML or higher. Large Pt islands enhance CO oxidation as well, but unlike small Pt islands, they also promote methanol dehydrogenation. Perturbations in electronic structure are responsible for the CO oxidation effect of defects, but the role of large Pt islands in promoting methanol dehydrogenation is primarily explained by surface geometric structure. PMID- 17129017 TI - Synthesis, characterization, and electrocatalytic activity of PtPb nanoparticles prepared by two synthetic approaches. AB - Intermetallic PtPb nanoparticles have been synthesized by two solution-phase reduction methods. In the first (PtPb-B), Pt and Pb salts were reduced by sodium borohydride in methanol at room temperature. In the second (PtPb-N), metal organic Pt and Pb precursors were reduced by sodium naphthalide in diglyme at 135 degrees C. Both methods produced small agglomerated nanoparticles of the ordered intermetallic PtPb (mean crystal domain size <15 nm) which were characterized by pXRD, SEM, UHV-STEM, BET, EDX, and electron diffraction. The electrocatalytic activity of PtPb nanoparticles produced by both methods toward formic acid and methanol oxidation was investigated and compared to Pt and PtRu. Both PtPb-B and PtPb-N nanoparticles exhibited enhanced electrocatalytic activity compared to commercially available Pt black and PtRu nanoparticles. For formic acid oxidation, the PtPb nanoparticles exhibited considerably lower onset potentials and higher current densities than Pt or PtRu. For methanol oxidation, the PtPb nanoparticles had onset potentials slightly positive of PtRu but exhibited higher current densities at potentials about 100 mV positive of onset. The general applicability of these methods for the synthesis of nanoparticles of ordered intermetallic phases is discussed. PMID- 17129018 TI - Morphology of platinum electrodeposits in the three-dimensional sublayer to full layer range produced under different potential modulations on highly oriented pyrolytic graphite. AB - The topography of platinum electrodes produced by electrodeposition (19 to 200 mC cm-2) on highly oriented pyrolytic graphite (HOPG) under different potential modulations was investigated by atomic force microscopy, scanning tunneling microscopy, and H-atom electrosorption voltammetry. To modulate electrodeposition, (i) triangular potential cycling at 0.1 V s-1, (ii) a linear cathodic potential at 0.1 V s-1 and anodic potential step cycling, and (iii) square wave potential cycling at 5000 Hz were utilized. AFM and STM imaging showed that at lower platinum loading the HOPG surface was partially covered by a 3D sublayer of platinum. Electrodes produced by procedure (i) were made of faceted platinum aggregates of about 200 nm and nanoclusters in the range of 5-20 nm; those that resulted from procedure (ii) consisted of anisotropic aggregates of nanoclusters arranged as quasi-parallel domains. These electrodes from (i) and (ii) behaved as fractal objects. The electrodes resulting from procedure (iii) exhibited a flat surface that behaved as a Euclidean object. For all WEs, as the platinum loading was increased the HOPG surface was fully covered by a thin 3D layer of platinum aggregates produced by electrodeposition and coalescence phenomena. Large platinum loading led to electrodes with fractal geometry. Statistical parameters (root-mean-square height, skewedness, kurtosis, anisotropy, Abbot curve, number of protrusions and valleys, and fractal dimension) were obtained from the analysis of AFM and STM imaging data. Platinum electrodeposition coupled to either H-adatom formation for procedures (i) and (ii) or phonon dispersion for (iii) was involved in the surface atom rearrangements related to electrofaceting. The H-adatom electrosorption voltammetry data were used to evaluate the real electrode surface area via the voltammetric charge and to advance a tentative explanation of the contribution of the different crystallographic facets to the global electrochemical process dominated by weak H-Pt adsorption interactions. PMID- 17129019 TI - Photochemically controlled electrochemical deposition and dissolution of Ag0 nanoclusters on au electrode surfaces. AB - A photoisomerizable thiolated nitrospiropyran SP, (1a), monolayer is assembled on a Au electrode by the primary deposition of thiolated nitromerocyanine isomer 1b as a monolayer on the electrode, followed by the irradiation of the surface with visible light, lambda > 475 nm. The surface coverage of nitrospiropyran units (1a) on the electrode is 2 x 10-10 mole cm-2. Irradiation of the electrode with UV light, 320 nm < lambda < 360 nm, results in the nitromerocyanine, MR, monolayer on the electrode that binds Ag+ ions to the phenolate units. The Ag+ ions associated with the MR monolayer undergo cyclic reduction to surface confined Ag0 nanoclusters, and reoxidation and dissolution of the Ag0 nanoclusters to Ag+ ions associated with the monolayer are demonstrated. The electron-transfer rate constants for the reduction of Ag+ to Ag0 and for the dissolution of Ag0 were determined by chronoamperometry and correspond to ketred = 12.7 s-1 and ketox = 10.5 s-1, respectively. The nanoclustering rate was characterized by surface plasmon resonance measurements, and it proceeds on a time scale of 10 min. The size of the Ag0 nanoclusters is in the range of 2 to 20 nm. The electrochemically induced reduction of the MR-Ag+ monolayer to the MR-Ag0 surface and the reoxidation of the MR-Ag0 surface control the hydrophilic hydrophobic properties of the surface. The advancing contact angle of the MR-Ag0 functionalized surface is 59 degrees , and the contact angle of the MR-Ag+ monolayer-functionalized surface is 74 degrees . Photoisomerization of the Ag0-MR surface to the Ag0-SP state, followed by the oxidation of the Ag0 nanoclusters, results in the dissolution of the Ag+ ions into the electrolyte solution. PMID- 17129020 TI - Cathodic electrodeposition of mixed molybdenum tungsten oxides from peroxo polymolybdotungstate solutions. AB - Mixed molybdenum tungsten trioxide films of varying stoichiometry (MoxW1 - xO3, 0 < x < 1) were prepared by cathodic electrodeposition on indium tin oxide (ITO) coated glass substrates from aqueous peroxo-polymolybdotungstate solutions. Electrochemical quartz crystal microbalance (EQCM), cyclic voltammetry, and chronocoulometry were used to gain insight into the electrodeposition mechanism. The compositional and structural properties were characterized for MoxW1 - xO3 films deposited at intermediate potentials (-0.35 V vs Ag/AgCl) and sintered at 250 degrees C using energy-dispersive spectroscopy, X-ray diffraction, and Raman spectroscopy. These studies reveal that films consist of homogeneously mixed MoxW1 - xO3, with an enriched Mo content ranging in composition from 0.4 < x < 0.7 depending upon the mol % Mo present in the deposition solution. Chronoamperometry and spectroelectrochemical measurements were conducted to estimate lithium ion diffusion coefficients and coloration efficiencies for the mixed metal oxide films in 1 M LiClO4/propylene carbonate. The subtle interplay between structural and compositional properties due to the uniform mixing of Mo and W oxide components shows that electrochromic and lithium ion transport properties are moderately enhanced relative to those of single-component WO3 and MoO3 and demonstrate improved structural stability over pure MoO3 polymorphs during electrochemical cycling. PMID- 17129021 TI - Electrocrystallization of Phase I, CuTCNQ (TCNQ = 7,7,8,8 Tetracyanoquinodimethane), on indium tin oxide and boron-doped diamond electrodes. AB - The electrochemical reduction of TCNQ to TCNQ*- in acetonitrile in the presence of [Cu(MeCN)4]+ has been undertaken at boron-doped diamond (BDD) and indium tin oxide (ITO) electrodes. The nucleation and growth process at BDD is similar to that reported previously at metal electrodes. At an ITO electrode, the electrocrystallization of more strongly adhered, larger, branched, needle-shaped phase I CuTCNQ crystals is detected under potential step conditions and also when the potential is cycled over the potential range of 0.7 to -0.1 V versus Ag/AgCl (3 M KCl). Video imaging can be used at optically transparent ITO electrodes to monitor the growth stage of the very large branched crystals formed during the course of electrochemical experiments. Both in situ video imaging and ex situ X ray diffraction and scanning electron microscopy (SEM) data are consistent with the nucleation of CuTCNQ taking place at a discrete number of preferred sites on the ITO surface. At BDD electrodes, ex situ optical images show that the preferential growth of CuTCNQ occurs at the more highly conducting boron-rich areas of the electrode, within which there are preferred sites for CuTCNQ formation. PMID- 17129022 TI - Attenuating electron-transfer rates via dendrimer encapsulation: the case of metal tris(bipyridine) core dendrimers. AB - Heterogeneous electron-transfer rates in metal tris(bipyridine) core dendrimers were measured using Osteryoung square-wave voltammetry. Rates decreased with generation, and this decrease could be correlated with the molecular weight increase. These results indicate that the coordination number around the redox center did not play any special role in sterically encapsulating the redox center. PMID- 17129023 TI - Voltammetry of suspensions of hollow particles with ferrocene-immobilized polyallylamine shells. AB - Redox-active hollow spheres were prepared through extracting a polystyrene core from the latex particle (PSPAAFc) composed of the core and the polyallylamine shell including ferrocenyl carboxylic amide. The suspension of the hollow spheres showed anodic and cathodic voltammetric peaks, which were nearly reversible and diffusion-controlled. The current was 3 times as large as the current for the suspension of the filled PSPAAFc. This value agreed with the theoretical one evaluated from the diameter (1.28 microm), the number of ferrocenyl moieties per particle, 1.2 x 108, by UV spectroscopy, and the diffusion coefficient obtained from the Stokes-Einstein equation. This fact indicates the reaction of the whole loaded charge, in contrast to the partial charge transfer of PSPAAFc. The dynamic flattening motion was observed to support the reaction of the whole charge. PMID- 17129024 TI - Study of peptide dendrimers having a ferrocene core supported on mercaptoundecanoic acid. AB - Hydrogen-bonding interactions between the carboxylic acid groups of mercaptoundecanoic acid (MUA) coated gold substrates and the ester surface of peptide dendrimers allows the formation of glutamic acid dendrimers films. Dendrimer films were prepared for generations 1-6 (G1-G6) and analyzed by spectroscopic and electrochemical techniques. Electrochemical studies using cyclic voltammetry and differential pulse voltammetry show that all films except those of G6 were electrochemically active. Lack of activity of G6 films is rationalized by the total encapsulation of the ferrocene redox probe by the dendritic sheath and lack of ion pairing, which prevents its oxidation. PMID- 17129025 TI - Polymer-modified opal nanopores. AB - The surface of nanopores in opal films, assembled from 205 nm silica spheres, was modified with poly(acrylamide) brushes using surface-initiated atom transfer radical polymerization. The colloidal crystal lattice remained unperturbed by the polymerization. The polymer brush thickness was controlled by polymerization time and was monitored by measuring the flux of redox species across the opal film using cyclic voltammetry. The nanopore size and polymer brush thickness were calculated on the basis of the limiting current change. Polymer brush thickness increased over the course of 26 h of polymerization in a logarithmic manner from 1.3 to 8.5 nm, leading to nanopores as small as 7.5 nm. PMID- 17129026 TI - Electrochemical template synthesis of multisegment nanowires: fabrication and protein functionalization. AB - Multisegment nanowires represent a unique platform for engineering multifunctional nanoparticles for a wide range of applications. For example, the optical and magnetic properties of nanowires can be tailored by modifying the size, shape, and composition of each segment. Similarly, surface modification can be used to tailor chemical and biological properties. In this article, we report on recent work on electrochemical template synthesis of nanogap electrodes, the fabrication of multisegment nanowires with embedded catalysts, and the selective functionalization of multisegment nanowires with proteins. PMID- 17129027 TI - Epitaxial electrodeposition of ZnO on Au(111) from alkaline solution: exploiting amphoterism in Zn(II). AB - The amphoteric nature of ZnO is used to produce the material from strongly alkaline solution. The solution pH is lowered globally to produce ZnO powder, and it is lowered locally at a Au(111) surface to produce epitaxial films. ZnO powder is precipitated from a solution of 10 mM Zn(II) in 0.25 M NaOH by simply adding 1 M HNO3 to the solution. For the film electrodeposition, the local pH at the electrode surface is decreased by electrochemically oxidizing the ascorbate dianion. The chemically precipitated ZnO powder grows with a sea urchin-like nanostructure, whereas the electrodeposited films have a columnar structure. ZnO electrodeposited onto a Au(111) single crystal has a ZnO(0001)[1011]//Au(111)[110] orientation relationship. PMID- 17129028 TI - New dendrimers containing a single cobaltocenium unit covalently attached to the apical position of Newkome dendrons: electrochemistry and guest binding interactions with cucurbit[7]uril. AB - Two new dendrimer series were prepared and characterized. These dendrimers contain a single bis(cyclopentadienyl)cobalt(III) (cobaltocenium, Cob+) unit covalently attached to the apical (focal) position of Newkome-type dendrons, ranging in size from first to third generation. The dendrimers in the first series (1ECob+-3ECob+) are hydrophobic and have 3, 9, and 27 tert-butyl esters on their peripheries, whereas the dendrimers in the second series (1Cob+-3Cob+) are hydrophilic with 3, 9, and 27 carboxylic acid groups on their surfaces, respectively. In voltammetric experiments, all dendrimers showed the expected one electron reversible reduction of the cobaltocenium center, and the heterogeneous rate of electron transfer decreased with generation in both dendrimer series. The host-guest binding interactions between water-soluble dendrimers 1Cob+-3Cob+ and the cucurbit[7]uril (CB7) host were investigated using 1H NMR spectroscopy, MALDI TOF mass spectrometry, and electrochemical techniques. The association equilibrium constants (K) for all dendrimer guests were significantly lower than that measured for the inclusion complex between underivatized Cob+ and CB7 (K = 5.7 x 10(9) M(-1)). Nonetheless, among the three dendrimers surveyed, the second generation dendrimer, 2Cob+, afforded optimum stabilization for the CB7 inclusion complex. PMID- 17129029 TI - Mechanistic study of the electrodeposition of nanoporous self-assembled ZnO/Eosin Y hybrid thin films: effect of eosin concentration. AB - ZnO films prepared by one-step electrodeposition in the presence of dissolved eosin molecules present an internal nanoporous hybrid structure resulting from self-assembling processes occurring in solution between ZnO and eosin components. This study aims to better understand the underlying growth mechanism, which is still unexplained. The films were deposited by cathodic electrodeposition from an oxygen-saturated aqueous zinc chloride solution. The effects of the addition of 10 to 100 micromol.L(-1) eosin Y, as a sodium salt, on the growth rate and film properties, were systematically studied while all other parameters remained constant (concentrations of zinc salt and supporting electrolyte, applied potential of -1.4 V versus the mercurous sulfate electrode (MSE), temperature of 70 degrees C, rotating disk electrode at 300 rotations per min, and a glass coated tin oxide electrode). It is shown that the addition of eosin provokes the formation of a nanoporous "cauliflower" structure whose nodule size and composition depend on the eosin concentration in the bath. The growth rate of the hybrid films increases markedly with the eosin concentration. The ZnO and eosin contents of the films are determined for each concentration by chemical analysis. Comparing with thickness determinations, it is shown that the total porosity increases up to 60-65% in volume fraction toward an eosin concentration of 100 micromol.L(-1). The empty pore volume fraction increases up to about 30% at an eosin concentration of about 20 micromol.L(-1) and then decreases. These correlations have been precisely established for the first time. It is shown that the global composition is fixed by the relative rate of deposition for zinc oxide, which is constant, and for the relative rate of eosin inclusion, which is proportional to the concentration in solution. This is explained on the basis of different steps in the growth mechanism, in particular, a diffusion effect limitation for both oxygen and eosin. This variation explains part of the increase in the growth rate. Another contribution is related by the structural effect on the nanoscale leading to the formation of the interpenetrated porous network. Competition between empty and eosin-filled parts of the pore network is evidenced. The formation of the porous network structure could be governed by a diffusion-limited aggregation mechanism. The system may represent a reference case of competing reactions in the electrochemical self-assembly of hybrid nanostructures. PMID- 17129030 TI - Elucidation of the redox behavior of 2,5-dimercapto-1,3,4-thiadiazole (DMcT) at poly(3,4-ethylenedioxythiophene) (PEDOT)-modified electrodes and application of the DMcT-PEDOT composite cathodes to lithium/lithium ion batteries. AB - The redox reactions of DMcT at PEDOT-modified glassy carbon electrodes (GCEs) in acetonitrile (AN) have been investigated via cyclic voltammetry (CV) and the electrochemical quartz crystal microbalance (EQCM) in order to elucidate the redox reaction mechanism. A redox couple at -0.29 V versus Ag/Ag+ was assigned to the dimerization process of singly protonated DMcT (DMcT-1H), and a second couple observed at +0.42 V was assigned to the polymerization process of the protonated DMcT dimer. Our investigations revealed further that the anodic current response at +0.55 V (polymerization process) has a shoulder at +0.38 V ascribed to the dimerization process of doubly protonated DMcT (DMcT-2H), indicating that the redox couple at +0.42 V is the overlapping response of the polymerization of the protonated DMcT dimer and the dimerization of the DMcT-2H monomer. It was also confirmed that the dimerization process of DMcT-1H at -0.29 V proceeded not only at the surface of a PEDOT film but also inside the film as previously suggested. Moreover, the thermodynamics of these redox reactions at PEDOT-modified GCEs are dependent on the basicity (or acidity) of the solution, as anticipated and previously shown at unmodified GCEs. The oxidation of DMcT occurs at less positive potentials and the reduction occurs at more negative potentials in the presence of base. On the basis of the results obtained, the full redox reaction scheme for DMcT at a PEDOT-modified GCE is proposed. PMID- 17129031 TI - Bismuth telluride (Bi2Te3) nanowires: synthesis by cyclic electrodeposition/stripping, thinning by electrooxidation, and electrical power generation. AB - Nanowires composed of the thermoelectric material Bi2Te3 were synthesized on highly oriented pyrolytic graphite (HOPG) electrodes using the electrochemical step edge decoration (ESED) method. Nanowire synthesis was initiated by applying a voltage pulse of -0.75 V versus SCE for 5 ms to an HOPG electrode in an aqueous solution containing both Bi3+ and TeO22-, thereby producing nuclei at the step edges. Bi2Te3 was electrodeposited onto these nuclei using a cyclic electrodeposition-stripping scheme that involved the electrodeposition of bismuth rich Bi2Te3 on a negative-going voltammetric scan (to -0.05 V) and the subsequent anodic stripping of excess bismuth from these nanowires during a positive-going scan (to +0.35 V). When this cycle was repeated 10-50 times, Bi2Te3 nanowires in the 100-300-nm-diameter range were obtained. These nanowires were narrowly dispersed in diameter (RSDdia = 10-20%), were more than 100 microm in length, and were organized into parallel arrays containing hundreds of wires. Smaller nanowires, with diameters down to 30 nm, were obtained by electrooxidizing 150-nm diameter Bi2Te3 nanowires at +0.37 V under conditions of kinetic control. This oxidation process unexpectedly improved the uniformity of Bi2Te3 nanowires, and X ray photoelectron spectroscopy (XPS) shows that these nanowires retain a Bi2Te3 core but also have a thin surface layer composed of Bi and Te oxides. The ability of Bi2Te3 nanowires to generate electrical power was assessed by transferring ensembles of these nanowires onto cyanoacrylate-coated glass surfaces and evaporating 4-point nickel contacts. A dimensionless figure of merit, ZT, ranging from 0 to 0.85 was measured for fresh samples that were less than 1 day old. XPS reveals that Bi2Te3 nanowires are oxidized within a week to Bi2O3 and TeO2. These oxides may interfere with the application by evaporation of electrical contacts to these nanowires. PMID- 17129032 TI - Electrochemical preparation and structural characterization of Co thin films and their anomalous IR properties. AB - Nanometer scale cobalt thin films of different structures and thicknesses supported on glassy carbon were prepared by electrochemical deposition under cyclic voltammetric conditions (denoted nm-Co/GC(n)). The thickness of Co thin films was altered systematically by varying the number (n) of potential cycling within a defined potential range in electrodeposition. Electrochemical in situ scanning tunneling microscopy (STM) and ex situ scanning electron microscopy (SEM) were employed to characterize the surface structure of Co thin films. It has been illustrated that the Co thin films were uniformly composed of Co nanoparticles, whose structure and size varied with increasing n. The structure of nanoparticles inside the Co thin films underwent a transition from bearded nanoparticles to multiform nanoparticles and finally to hexagonal nanosheets, accompanying with an increase of average size. In situ FTIR reflection spectroscopic studies employing CO adsorption as probe reaction revealed that the Co thin films all exhibited anomalous IR properties; that is, along with their different nanostructures they presented abnormal IR effects, Fano-like IR effects, and surface-enhanced IR absorption effects. CO adsorbed on Co thin films dominated by bearded nanoparticles yielded abnormal IR absorption bands; that is, the direction of the bands is inverted completely, with enhanced intensity in comparison with those of CO adsorbed on a bulk Co electrode. The enhancement of abnormal IR absorption has reached a maximal value of 26.2 on the nm-Co/GC(2) electrode. Fano-like IR features, which describe the bipolar IR bands with their positive-going peak on the low wavenumbers side, were observed in cases of CO adsorbed on Co thin films composed mainly of multiform nanoparticles, typically on the nm-Co/GC(8) electrode. IR features were finally changed into surface enhanced IR absorption as CO adsorbed on the nm-Co/GC(30) electrode, on which the Co thin film is dominated by Co hexagonal nanosheets. PMID- 17129033 TI - Effect of peptide ligand dipole moments on the redox potentials of Au38 and Au140 nanoparticles. AB - Phenylethanethiolate monolayer-protected Au38 and Au140 nanoclusters were modified by ligand place exchange with a series of thiolated peptides. The peptides were homooligomers based on the alpha-aminoisobutyiric acid unit. The effects of changing the peptide concentration and the peptide length in the capping monolayer were studied by differential pulse voltammetry. The results showed that the redox behavior of the nanoparticles can be affected very significantly by such modifications. For example, the first oxidation peak of Au38, a cluster displaying molecule-like behavior, could be shifted positively by as much as 0.7-0.8 V. Detectable redox shifts were noted even when one single oriented peptide was in the Au140 monolayer. These effects were attributed to the molecular dipole moments of the peptide ligands. PMID- 17129034 TI - Preliminary studies in the electrodeposition of PbSe/PbTe superlattice thin films via electrochemical atomic layer deposition (ALD). AB - This paper concerns the electrochemical growth of compound semiconductor thin film superlattice structures using electrochemical atomic layer deposition (ALD). Electrochemical ALD is the electrochemical analogue of atomic layer epitaxy (ALE) and ALD, methods based on nanofilm formation an atomic layer at a time, using surface-limited reactions. Underpotential deposition (UPD) is a type of electrochemical surfaced-limited reaction used in the present studies for the formation of PbSe/PbTe superlattices via electrochemical ALD. PbSe/PbTe thin-film superlattices with modulation wavelengths (periods) of 4.2 and 7.0 nm are reported here. These films were characterized using electron probe microanalysis, X- ray diffraction, atomic force microscopy (AFM), and infrared reflection absorption measurements. The 4.2 nm period superlattice was grown after deposition of 10 PbSe cycles, as a prelayer, resulting in an overall composition of PbSe0.52Te0.48. The 7.0 nm period superlattice was grown after deposition of 100 PbTe cycle prelayer, resulting for an overall composition of PbSe0.44Te0.56. The primary Bragg diffraction peak position, 2theta, for the 4.2 superlattice was consistent with the average (111) angles for PbSe and PbTe. First-order satellite peaks, as well as a second, were observed, indicating a high-quality superlattice film. For the 7.0 nm superlattice, Bragg peaks for both the (200) and (111) planes of the PbSe/PbTe superlattice were observed, with satellite peaks shifted 1 degrees closer to the (111), consistent with the larger period of the superlattice. AFM suggested conformal superlattice growth on the Au on glass substrate. Band gaps for the 4.2 and 7.0 nm period superlattices were measured as 0.48 and 0.38 eV, respectively. PMID- 17129035 TI - Conducting metallopolymers based on azaferrocene. AB - A series of 2,5-thiophene-substituted 1',2',3',4',5'-pentamethylazaferrocene complexes were synthesized and electropolymerized to produce polymers with fully pi-conjugated backbones. The length and hence oxidation potential of the conjugated linker (the thiophene fragments) between the metal centers were varied to understand the influence of the metal-metal interactions on the overall electroactivity of the resulting polymer. These complexes were electrochemically polymerized, and the resulting polymers were characterized by cyclic voltammetry, in situ conductivity, and spectroelectrochemistry measurements. The iron-centered oxidations significantly increased the conductivity of the polymer. The results reveal that shorter conjugated linkers cause the onset of conductivity to occur at lower potentials. This effect implies that a superexchange mechanism is likely operative in the charge migration of these polymers. PMID- 17129036 TI - Redox-active silica nanoparticles. Part 1. Electrochemistry and catalytic activity of spherical, nonporous silica particles with nanometric diameters and covalently bound redox-active modifications. AB - Nonporous spherical silica nanoparticles resulting from a controlled Stober process are covalently surface modified with redox-active molecules. Ferrocene, a ruthenium(II) complex with an N2P2Cl2 ligand set, and a sterically hindered biphenylamine are used as modifiers. The resulting materials are characterized by physical, spectroscopic, electrochemical, and chemical methods. The cyclic voltammetric behavior is studied in detail and reveals effects of charge transport by electron hopping along the surface of particles adsorbed on a Pt electrode. The ruthenium(II) complex remains catalytically active with respect to hydrogenation upon immobilization on the particles. Thus, the respective material provides a heterogenized homogeneous hydrogenation catalyst on a solid support. PMID- 17129037 TI - Influence of electrode rotation on the growth and impedance of a low band gap conducting polymer. AB - Rotation of the electrode during the electrochemical polymerization of delta4,4' di-cyclopenta[2,1-b;3',4'-b']-dithiophene results in enhanced rates of film growth on the electrode and changes in morphology from dominantly fibrilar to globular structures. The impedance of the resulting films shows their ionic conductivities to be higher than their electronic conductivities. Rotating the electrode during growth enhances electronic conductivities by as much as 2 orders of magnitude, and this is attributed to the dominance of growth of the polymer on the electrode surface (grafting) over the precipitation of material from the diffusion layer. PMID- 17129038 TI - Self-assembly of C(60) pi-extended tetrathiafulvalene (exTTF) dyads on gold surfaces. AB - The first self-assembly of a C60 pi-extended tetrathiafulvalene (exTTF) dyad on a gold surface is reported. Four fullerene derivatives, two of them containing p quinonoid pi-extended tetrathiafulvalenes (exTTFs), have been synthesized, and their solution electrochemistry has been investigated by means of cyclic voltammetry. Fullerene-containing SAMs of thioctic acid derivatives 3 and 6 have also been investigated by cyclic voltammetry. The cyclic voltammograms of both compounds exhibit three reversible reduction waves, and for compound 6, one irreversible oxidation process corresponding to the oxidation of the exTTF subunit is observed. Stable self-assembled monolayers (SAMs) of fullerene derivative 3 were formed on gold surfaces, whereas dyad 6 does not present a very clear electrochemical response, most probably as a result of structural rearrangements on the monolayer or charge transfer between the C60 and exTTF moieties. PMID- 17129039 TI - Electrochemical synthesis of dendritic zinc films composed of systematically varying motif crystals. AB - Polycrystalline zinc films with new dendritic frameworks were electrodeposited from nonaqueous formamide media containing 0.01-0.3 M Zn(ClO4)2.6H2O as the Zn2+ source and 0.1 M LiClO4.3H2O as the supporting electrolyte. Formamide media offer a wider range of deposition temperatures and deposition potentials than aqueous solutions, which provides a higher degree of freedom in creating new polycrystalline architectures. The growth patterns of zinc crystals could be precisely controlled (e.g., faceted growth and dendritic growth) by changing the interplay between the growth rate and the mass transport rate. The effect of deposition potential, temperature, and Zn2+ concentration on the onset potential of dendritic growth and the detailed dendritic features were studied systematically. The zinc dendrites obtained in this study were composed of submicron-sized crystals of a uniform shape (motif crystals) that grow repetitively fused together to form three-dimensionally dispersed dendritic backbones. This unique organization achieves a remarkable physical and electrical continuity between crystals while generating high surface areas, which is difficult to accomplish simultaneously in polycrystalline films. The shape of motif crystals can be finely tuned from hexagons to fern-shaped leaves by the deposition potential applied, which in turn alters the overall degree of branching of dendritic backbones. Cyclic voltammetry of the resulting zinc electrodes with various growth patterns was carried out and discussed in conjunction with the films' morphological variation. PMID- 17129040 TI - Planar electric double layer for a restricted primitive model electrolyte at low temperatures. AB - Monte Carlo simulation and the modified Poisson-Boltzmann theory are used to investigate the planar electric double layer for a restricted primitive model electrolyte at low temperatures. Capacitance as a function of temperature at low surface charge is determined for 1:1, 2:2, 2:1, and 3:1 electrolytes. Negative adsorption can occur for 1:1 electrolytes at low surface charge with low electrolyte concentration. The 1:1 electrolyte diffuse layer potential as a function of surface charge displays a maximum at low densities. At high densities, the diffuse layer potential is negative with a negative slope. The Gouy-Chapman-Stern theory fails in this low-temperature regime, whereas the modified Poisson-Boltzmann theory is fairly successful in this regard. PMID- 17129041 TI - New developments in the theory of the diffuse double layer. AB - The role of ion size effects in determining diffuse layer properties is considered. Monte Carlo data relevant to this question are reviewed. Then the integral equation approach to the problem is considered with an emphasis on attempts to derive an analytical equation for the potential drop across the diffuse layer. An empirical model for ion size effects is described that allows one to estimate not only the diffuse layer potential drop but also the diffuse layer capacity, the ionic surface excesses, and the potential distribution in the diffuse layer. It is demonstrated that the resulting model can easily be applied by experimentalists to the analysis of experimental data for double layer phenomena in electrochemistry and colloid science. PMID- 17129042 TI - Potential-, pH-, and isotope-dependence of proton-coupled electron transfer of an osmium aquo complex attached to an electrode. AB - An osmium complex, [OsII(bpy)2(4-aminomethylpyridine)(H2O)]2+, is attached to a mixed self-assembled monolayer on a gold electrode. The complex exhibits 1 electron, 1-proton redox chemistry (OsIII(OH)/OsII(H2O)) at pHs and potentials that are experimentally accessible with gold electrodes in aqueous electrolytes. The thermodynamic behavior and kinetic behavior of the system are investigated as a function of pH in both H2O and D2O. The two formal potentials and two pKa values are relatively constant for two chain lengths in H2O and in D2O. The standard rate constants at all pHs are strongly and uniformly affected by chain length, indicating that electronic coupling is the dominant factor controlling the rate of electron transfer. In both H2O and D2O, the standard rate constant is weakly dependent on the pH, exhibiting a minimum value midway between the pKa values. The kinetic isotope effect is small; standard rate constants decrease by roughly a factor of 2 in D2O over a wide range of pHs, but not at the more acidic pHs. The Tafel plots and plots of the transfer coefficient vs overpotential are asymmetrical at all pHs. These results are interpreted in terms of a larger reorganization energy for the OsII species and a smaller reorganization energy for the OsIII species. The OsIII reorganization energy is constant at all pHs in both H2O and D2O. The pH dependence of the OsII reorganization energy accounts for some or all of the pH dependence of the standard rate constant in H2O and D2O. The data deviate substantially from predictions of the stepwise proton coupled electron-transfer mechanism. The observation of a kinetic isotope effect supports the concerted mechanism. PMID- 17129043 TI - CdSe sensitized thin aqueous films: probing the potential distribution inside multilayer assemblies. AB - Ultrathin polypeptide multilayer films are assembled by the sequential electrostatic adsorption of monolayers of poly-l-lysine and poly-l-glutamic acid onto carboxylic acid terminated alkanethiol-modified gold surfaces. The polypeptide multilayer films are hydrophilic, can incorporate electroactive species such as ferri/ferrocyanide, and are stable when immersed in organic solvents such as 1,2-dichloroethane. Cadmium selenide quantum dots stabilized by negatively charged citrate groups are electrostatically attached to the multilayer film assembly in order to act as photoactive species. Photocurrent responses originating from the CdSe sensitized ultrathin multilayer film are investigated as functions of the applied potential, the thickness of the film and the presence of quenchers in the organic phase. A theoretical model is proposed in order to analyze the kinetics of the photoinduced electron-transfer reactions and to probe the potential distribution within the film. PMID- 17129044 TI - Theoretical approach to ion penetration into pores with pore fractal characteristics during double-layer charging/discharging on a porous carbon electrode. AB - The effects of pore fractal characteristics on the kinetics of double-layer charging/discharging on a porous carbon electrode were investigated by using theoretical calculations of potentiostatic current transients (PCTs) and cyclic voltammograms (CVs). Prior to theoretical calculation, it was experimentally evidenced that pore fractality is clearly possessed by the porous carbon electrode. From the analyses of the PCTs and the CVs theoretically calculated at various values of pore fractal dimension dF,pore, inner cutoff length rmin, and outer cutoff length rmax of the pore fractality, it was found that as dF,pore increased, the absolute values of the derivatives of the logarithmic PCTs decreased to 0.5, and the current decayed more slowly with time. The rate capability gamma decreases with increasing dF,pore over the whole scan-rate range, which leads to the lower power density. As rmin increased, the current decayed more rapidly in the later stage of the PCT, which is mainly limited by the smaller pores. On the other hand, as rmax increased, the current decayed more rapidly in the earlier stage of the PCT, which is mainly determined by the larger pores. Moreover, the larger values of rmin and rmax enhance the rate capability gamma as well, but they reduce the double-layer capacitance. The beneficial contribution of the larger pores to the power density competes with the detrimental contribution of those pores to the energy density. PMID- 17129045 TI - Adaptive finite element methods in electrochemistry. AB - In this article, we review some of our previous work that considers the general problem of numerical simulation of the currents at microelectrodes using an adaptive finite element approach. Microelectrodes typically consist of an electrode embedded (or recessed) in an insulating material. For all such electrodes, numerical simulation is made difficult by the presence of a boundary singularity at the electrode edge (where the electrode meets the insulator), manifested by the large increase in the current density at this point, often referred to as the edge effect. Our approach to overcoming this problem has involved the derivation of an a posteriori bound on the error in the numerical approximation for the current that can be used to drive an adaptive mesh generation algorithm, allowing calculation of the quantity of interest (the current) to within a prescribed tolerance. We illustrate the generic applicability of the approach by considering a broad range of steady-state applications of the technique. PMID- 17129046 TI - Studies of potential inversion in an extended tetrathiafulvalene. AB - Electrochemical oxidation of the extended tetrathiafulvalene 9,10-bis(1,3 dithiole-2-ylidene)-9,10-dihydroanthracene (2) was studied in N,N dimethylformamide. A single, two-electron oxidation peak occurs, and on the return sweep of a cyclic voltammogram, a two-electron reduction peak is seen. The oxidation of 2 to its cation radical and dication occurs with potential inversion (i.e., removal of the second electron occurs more easily than removal of the first). The extent of potential inversion was estimated by cyclic voltammetry to be 0.28 V by analysis of the process in terms of concerted structural change and electron transfer. Failure to detect the cation radical by EPR of an equimolar mixture of neutral 2 and the dication is consistent with this value. The inner reorganization energy of the cation radical was determined by gas-phase photoelectron spectroscopy (PES) to be 0.31-0.35 eV. Calculations, consistent with earlier experimental data, show rather large changes in structure associated with the oxidation processes. These large structural changes contrast with the relatively small inner reorganization energy found by PES. This observation prompted an analysis of voltammetry in terms of two-step processes, with structural change either preceding or following electron transfer. Agreement of simulations based on this mechanism with experimental voltammograms was equally as good as with the concerted mechanism. Notably, the two-step mechanism produced more realistic values of the transfer coefficient and electron-transfer rate constant for the first step of oxidation. PMID- 17129047 TI - Redox-driven conductance switching via filament formation and dissolution in carbon/molecule/TiO2/Ag molecular electronic junctions. AB - Carbon/molecule/TiO2/Au molecular electronic junctions show robust conductance switching, in which a metastable high conductance state may be induced by a voltage pulse which results in redox reactions in the molecular and TiO2 layers. When Ag is substituted for Au as the "top contact", dramatically different current/voltage curves and switching behavior result. When the carbon substrate is biased negative, an apparent breakdown occurs, leading to a high conductance state which is stable for at least several hours. Upon scanning to positive bias, the conductance returns to a low state, and the cycle may be repeated hundreds of times. Similar effects are observed when Cu is substituted for Au and for three different molecular layers as well as "control" junctions of the type carbon/TiO2/Ag/Au. The polarity of the "switching" is reversed when the Ag layer is between the carbon and molecular layers, and the conductance change is suppressed at low temperature. Pulse experiments show very erratic transitions between high and low conductivity states, particularly near the switching threshold. The results are consistent with a switching mechanism based on Ag or Cu oxidation, transport of their ions through the TiO2, and reduction at the carbon to form a metal filament. PMID- 17129048 TI - Electrochemistry of TEMPO in the aqueous liquid/vapor interfacial region: measurements of the lateral mobility and kinetics of surface partitioning. AB - A new method is described to simultaneously determine the kinetics of surface partitioning and the lateral diffusion constant of redox active amphiphiles. It concerns water-soluble amphiphiles for which the surface adsorption equilibrium constant and the solution diffusion constant are measured independently. The method involves cyclic voltammetric experiments carried out at the air/water interface with microband electrodes aligned with the plane of the water surface. Typically, 100 nm wide, 1.0 cm long microband electrodes are fabricated by the vacuum vapor deposition of gold films on glass. The front face of the electrode substrates are coated with impermeable, dimensionally stable, polymer barrier films with thickness L in the range of approximately 0.1-1.0 microm. Fracturing such gold-coated glass substrates exposes gold microbands. The recorded voltammetric current sensitively depends on the barrier film thickness, the surfactant surface diffusion constant, Dsurf, and its rate constant of desorption, kdes. For a given surfactant, such as the nitroxyl piperidine free radical TEMPO featured in this report, large currents are observed with microband electrodes that do not carry a barrier film (L = 0). This is because the surfactant surface population diffusing along the air/water interface can be directly electro-oxidized at the edge of the microband. Smaller currents are measured in the presence of a barrier film, since, in those instances, the surface population may contribute to the voltammetric current only via a mechanism involving surfactant desorption from the water surface into bulk, where it contributes to the three-dimensional solution diffusion processes. The quantitative interpretation of the voltammetric experiments was made possible with finite element simulations with FEMLAB. These produce a set of calibration curves, Dsurf versus log kdes, for each value of the barrier film thickness. The intersection of the calibration curves determines the unique values of Dsurf and kdes. For TEMPO, Dsurf = 4.4 +/- 1.2 x 10(-5) cm2/s and kdes >/= 2 x 10(4) s(-1). Surfactant desorption rate constants of this magnitude have not been previously experimentally accessible. Since, in our earlier report (Wu, D. G.; Malec, A. D.; Head-Gordon, M.; Majda, M. J. Am. Chem. Soc. 2005, 27, 4490-4496), we showed that TEMPO is not immersed in water and that it diffuses along the interface hydrogen bonded to just one or two water molecules, its Dsurf value approximates the water diffusion constant in the aqueous liquid-vapor interfacial region. PMID- 17129049 TI - SECM study of solute partitioning and electron transfer at the ionic liquid/water interface. AB - Molecular partitioning and electron-transfer kinetics have been studied at the ionic liquid/water (IL/water) interface by scanning electrochemical microscopy (SECM). The ionic liquid C8mimC1C1N is immiscible with water and forms a nonpolarizable interface when in contact with it. Partitioning of ferrocene (Fc) across the IL/water interface was studied by SECM and found to be kinetically fast with a partition coefficient CIL/CW of 2400:1. The partition coefficient value was measured by SECM under quasi-steady-state conditions without waiting for complete solute equilibration. To investigate the kinetics of the electron transfer (ET) between aqueous ferricyanide and Fc dissolved in IL, a new approach to the analysis of the SECM current-distance curves was developed to separate the contributions of Fc partitioning and the ET reaction to the tip current. Several combinations of different aqueous and nonaqueous redox species were investigated; however, only the Fc/Fe(CN)63- system behaved according to the Butler-Volmer formalism over the entire accessible potential range. PMID- 17129050 TI - Fabrication and functionalization of nanochannels by electron-beam-induced silicon oxide deposition. AB - We report on the fabrication and electrical characterization of functionalized solid-state nanopores in low stress silicon nitride membranes. First, a pore of approximately 50 nm diameter was drilled using a focused ion beam technique, followed by the local deposition of silicon dioxide. A low-energy electron beam induced the decomposition of adsorbed tetraethyl orthosilicate resulting in site selective functionalization of the nanopore by the formation of highly insulating silicon oxide. The deposition occurs monolayer by monolayer, which allows for control of the final diameter with subnanometer accuracy. Changes in the pore diameter could be monitored in real time by scanning electron microscopy. Recorded ion currents flowing through a single nanopore revealed asymmetry in the ion conduction properties with the sign of the applied potential. The low frequency excess noise observed at negative voltage originated from stepwise conductance fluctuations of the open pore. PMID- 17129051 TI - Stability of the gold/silica thin film interface: electrochemical and surface plasmon resonance studies. AB - This article reports chemical stability studies of a gold film electrode coated with thin silicon oxide (SiOx) layers using electrochemical, surface plasmon resonance (SPR) and atomic force microscopy (AFM) techniques. Silica films with different thicknesses (d = 6.4, 9.7, 14.5, and 18.5 nm) were deposited using a plasma-enhanced chemical vapor deposition technique (PECVD). For SiOx films with d >/= 18.5 nm, the electrochemical behavior is characteristic of a highly efficient barrier for a redox probe. SiOx films with thicknesses between 9.5 and 14.5 nm were found to be less efficient barriers for electron transfer. The Au/SiOx interface with 6.4 nm of SiOx, however, showed an enhanced steady-state current compared to that of the other films. The stability of this interface in solutions of different pH was investigated. Whereas a strongly basic solution led to a continuous dissolution of the SiOx interface, acidic treatment produced a more reticulated SiOx film and improved electrochemical behavior. The electrochemical results were corroborated by SPR measurements in real time and AFM studies. PMID- 17129052 TI - Kinetics of the adhesion of DMPC liposomes on a mercury electrode. Effect of lamellarity, phase composition, size and curvature of liposomes, and presence of the pore forming peptide mastoparan X. AB - Liposomes suspended in aqueous electrolyte solutions can adhere at mercury electrodes. The adhesion is a complex process that starts with the docking and opening and leads to a spreading, finally resulting in the formation of islands of adsorbed lecithin molecules. The adhesion process can be followed by chronoamperometry, and a detailed analysis of the macroscopic and microscopic kinetics can be performed yielding rate constants and activation parameters. By using giant unilamellar liposomes and multilamellar liposomes, the effect of lamellarity and liposome size could be elucidated for liposomes in the liquid crystalline, gel, and superlattice phase states. Below the phase transition temperature, the time constant of opening of the liposomes (i.e., the irreversible binding of the lecithin molecules on the preliminary contact interface liposome|mercury and the therewith associated disintegration of the liposome membrane on that spot) is shown to be strongly size dependent. The activation energy, however, of that process is size independent with the exception of very small liposomes. That size dependence of time constants is a result of the size dependence of the initial contact area. The time constant and the activation energies of the spreading step exhibit a strong size dependence, which could be shown to be due to the size dependence of rate and activation energy of pore formation. Pore formation is necessary to release the solution included in the liposomes. This understanding was corroborated by addition of the pore inducing peptide Mastoparan X to the liposome suspension. The obtained results show that electrochemical studies of liposome adhesion on mercury electrodes can be used as a biomimetic tool to understand the effect of membrane properties on vesicle fusion. PMID- 17129053 TI - Selective anion sensing based on tetra-amide calix[6]arene derivatives in solution and immobilized on gold surfaces via self-assembled monolayers. AB - Two anion receptors, 1 and 2, based on the calix[6]crown-4 architecture were synthesized and characterized by NMR (1H, 13C, COSY), UV-vis, and MALDI-MS. 1H NMR measurements demonstrate that receptors 1 and 2 exhibit the highest binding affinity for fluoride ions compared to other anions including Cl-, Br-, NO3-, HSO4-, H2PO4-, and AcO-. The binding constants of 1 with F- and AcO- are 326 (+/ 32) and 238 (+/-23) M-1, whereas those of 2 with F- and AcO- are 222 (+/-25) and 176 (+/-21) M-1. The fluorescent titration of 2 with various anions such as Cl-, Br-, NO3-, HSO4-, and H2PO4- led to essentially no change in excimer emission and a slight enhancement of monomer emission. In contrast, a dramatic change was observed in the fluorescence spectra upon the addition of F- and AcO- to 2. Self assembled monolayers (SAMs) of 1 were formed on gold surfaces and characterized by reductive desorption and other techniques. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy were used to monitor anion recognition by the SAM-modified gold electrodes. The gold electrodes modified by SAMs of 1, upon binding with the F- anion, exhibit a dramatic increase in charge-transfer resistance (Rct) values. This is due to the repulsion between the negatively charged electrode surfaces and the negatively charged Fe(CN)6(3-/4-) redox probe in the electrolyte solution. In contrast, smaller increases in Rct values were observed in the cases of other monovalent anions investigated. PMID- 17129054 TI - Microscale patterning of organic films on carbon surfaces using electrochemistry and soft lithography. AB - We have demonstrated three simple strategies employing poly(dimethylsiloxane) (PDMS) molds for patterning carbon surfaces with two different modifiers in an 18 microm line pattern. The PDMS molds are patterned with microfluidic channels (approximately 22 microm wide and 49 microm deep) and form a reversible, conformal seal to the pyrolyzed photoresist film (PPF) and modified PPF surfaces. Modifiers are electrochemically grafted to the PPF surface by the reduction of aryl diazonium salts and the oxidation of primary amines. For the fill-in patterning approach, the first modifier is electrografted to the PPF surface exposed within the microchannels, and in a second grafting step after removal of the PDMS mold, the second modifier fills in the remaining surface. The selective conversion strategy involves electrografting a continuous film of the modifier to the PPF surface, sealing the PDMS mold to the modified surface and carrying out an irreversible electrochemical reaction of the modifier exposed within the microchannels. In the build-up patterning approach, the PDMS mold is sealed to the modified PPF surface, and a chemical coupling reaction is effected in the microchannels to build up the pattern. The patterns are characterized using SEM, optical microscopy, the formation of condensation figures, and SEM imaging after the assembly of Au nanoparticles. PMID- 17129055 TI - Electrochemical grafting of an aryl fluorosulfonimide electrolyte onto glassy carbon. AB - Robust bonded layers of an aromatic fluorosulfonimide electrolyte were created on glassy carbon disk and plate electrodes by electroreduction of a new fluorosulfonimide aryl diazonium zwitterion. Formation of the bonded layer was confirmed by XPS of the modified surfaces and also by the effect of the bound layer on reduction/oxidation of redox-active probe molecules. Surface coverage in the monolayer range was achieved. Counterions for as-formed layers were initially tetra-alkylammonium ions from the coating electrolyte but could subsequently be exchanged for other cations by exposure to salt solutions. The bonded layers are very stable with respect to treatment with solvents (water and/or acetonitrile), dry heat (120 degrees C), and hot acid (triflic acid, 80 degrees C, neat and containing 50 wt % water); however, they are unstable with respect to electro oxidative scanning in aqueous electrolyte solutions. PMID- 17129056 TI - Photonic and electrochemical properties of adsorbed [Ru(dpp)2(Qbpy)]2+ luminophores. AB - Dense monolayers of [Ru(dpp)2Qbpy]2+, where dpp is 4,4'-diphenylphenanthroline and Qbpy is 2,2':4,4' ':4'4' '-quarterpyridyl, have been formed by spontaneous adsorption onto clean platinum microelectrodes. The cyclic voltammetry of these monolayers is nearly ideal, and three redox states are accessible over the potential range of +/-1.3 V. Chronoamperometry conducted on the microsecond time scale has been used to probe the dynamics of heterogeneous electron transfer and indicates that the standard heterogeneous electron-transfer rate constant, k degrees , is approximately 106 s-1. The metal complex emits at approximately 600 nm in fluid and solid solution as well as when bound to a platinum electrode surface within a dense monolayer. In the case of the monolayers, it appears that the excited states are not completely deactivated by radiationless energy transfer to the metal because electronic coupling between the adsorbates and the electrode is weak. The dynamics of lateral electron transfer between the electronically excited Ru2+* and ground-state Ru3+ species has been explored by measuring the luminescence intensity after defined quantities of Ru3+ have been produced electrochemically within the monolayer. The rate of lateral electron transfer is between 8 x 106 and 3 x 108 M-1 s-1, indicating efficient electron transfer between adsorbates in close-packed assemblies. Voltammetry conducted at megavolt per second scan rates has been used to directly probe the redox properties of the electronically excited species. PMID- 17129057 TI - Molecular adsorption at well-defined electrode surfaces: hydroquinone on Pd(111) studied by EC-STM. AB - The interaction of hydroquinone (H2Q) with well-defined Pd(111) surfaces at preselected potentials in dilute H2SO4 has been studied by molecule-resolved electrochemical scanning tunneling microscopy (EC-STM). H2Q spontaneously undergoes oxidative chemisorption to benzoquinone (Q), which adopts a slightly tilted parallel orientation. Evidently, the surface coordination is through the quinone pi-electron system. At potentials within the double-layer region, a close packed well-ordered Pd(111)-(3 x 3)-Q adlattice was formed. A potential excursion to 0.7 V, a potential at which the solution-phase Q/H2Q redox reaction takes place, introduced disorder into the organic adlayer; this positive-potential induced order-to-disorder phase transition is reversible because the ordered (3 x 3)-Q adlattice was regenerated when the potential reverted to 0.4 V. When the potential was poised at 0.2 V, a potential at which hydrogen evolution was initiated, an appreciable fraction of Q was (hydrogenatively) desorbed; the remnant Q molecules were agglomerated in small islands that retained the (3 x 3) symmetry of the full adlayer. Two possible structural models of the Pd(111)-(3 x 3)-Q adlattice are described. PMID- 17129058 TI - Electrochemical control of the structure of two-dimensional supramolecular organization consisting of phthalocyanine and porphyrin on a gold single-crystal surface. AB - Two-component adlayers consisting of cobalt(II) phthalocyanine (CoPc) and a metalloporphyrin such as 5,10,15,20-tetraphenyl-21H,23H-porphine copper(II) (CuTPP), 2,3,7,8,12,13,17,18-octaethyl-21H,23H-porphine copper(II) (CuOEP), or 5,10,15,20-tetraphenyl-21H,23H-porphine cobalt(II) (CoTPP) were prepared by immersing either an Au(111) or Au(100) substrate in a benzene solution containing those molecules. The mixed adlayers thus prepared were investigated in 0.1 M HClO4 by cyclic voltammetry (CV) and in situ scanning tunneling microscopy (STM). The composition of the mixed adlayer consisting of CoPc and CuTPP molecules was found to vary with immersion time. CoPc molecules displaced CuTPP molecules during the modification process with increasing immersion time, and the CuTPP molecules were completely displaced by CoPc molecules in the mixed solution after a prolonged modification time, during which the underlying Au(100) substrate underwent phase transition from the reconstructed (hex) lattice to the unreconstructed (1 x 1) lattice. The two-component adlayer of CoPc and CuTPP was found to form a supramolecular adlayer with the constituent molecules arranged alternately on Au(100)-(hex). The striped structure was stable on Au(100)-(hex) at or near the open circuit potential (OCP), whereas the mixed adlayer was disordered on Au(100)-(1 x 1) at potentials more positive than OCP, where the phase transition of the arrangement of underlying Au atoms (i.e., the lifting of reconstruction) was induced electrochemically. A similar two-component supramolecular adlayer consisting of CoPc and CuTPP was formed on Au(111). A highly ordered, compositionally disordered adlayer of CoTPP and CuTPP was formed on Au(100)-(hex), suggesting that the adlayer structure is independent of the coordinated central metal ion for the formation of supramolecular nanostructures composed of those molecules. A supramolecular organization of CoPc and CuOEP was also found on Au(111). The surface mobility and the molecular reorganization of CoPc and CuOEP on Au(111) were tuned by modulation of the electrode potential. It is concluded that molecular assemblies of the two-component structure consisting of phthalocyanine and porphyrin were controlled not only by the crystallographic orientation of Au but also by the modulation of electrochemical potential. PMID- 17129059 TI - Ionic conductivity of the aqueous layer separating a lipid bilayer membrane and a glass support. AB - The in-plane ionic conductivity of the approximately 1-nm-thick aqueous layer separating a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) bilayer membrane and a glass support was investigated. The aqueous layer conductivity was measured by tip-dip deposition of a POPC bilayer onto the surface of a 20- to 75 microm-thick glass membrane containing a single conical-shaped nanopore and recording the current-voltage (i-V) behavior of the glass membrane nanopore/POPC bilayer structure. The steady-state current across the glass membrane passes through the nanopore (45-480 nm radius) and spreads radially outward within the aqueous layer between the glass support and bilayer. This aqueous layer corresponds to the dominant resistance of the glass membrane nanopore/POPC bilayer structure. Fluorescence recovery after photobleaching measurements using dye-labeled lipids verified that the POPC bilayer maintains a significant degree of fluidity on the glass membrane. The slopes of ohmic i-V curves yield an aqueous layer conductivity of (3 +/- 1) x 10(-3) Omega(-1) cm(-1) assuming a layer thickness of 1.0 nm. This conductivity is essentially independent of the concentration of KCl in the bulk solution (10-4 to 1 M) in contact with the membrane. The results indicate that the concentration and mobility of charge carriers in the aqueous layer between the glass support and bilayer are largely determined by the local structure of the glass/water/bilayer interface. PMID- 17129060 TI - Patterning cellular motility using an electrochemical technique and a geometrically confined environment. AB - We describe herein a method for controlling the pattern of permissible cell migration and proliferation on a substrate in time and space. Using this method, a confluent monolayer of cells that is confined within a defined region is released into a neighboring region. Incorporated into the method is an electrochemical technique that uses a scanning microelectrode to draw regions on the surface of the system that thereafter can support cell migration and growth. The supporting glass substrate is patterned with regions of 2 methacryloyloxyethyl phosphorylcholine (MPC) polymer that are not affected by the electrochemical treatment and also robustly resist cellular overgrowth as well as regions that can be individually switched when electrochemically treated from cell repellent to cell adhering. It is therefore possible to strictly define the areas into which cells can migrate. We found that HeLa cells migrate more rapidly as the width of cell-adhering lanes increases until a width of ca. 50 microm is reached, at which point the migration rate is roughly constant. We also designed a drug assay using our cell migration technique. The technique allows for cell migration only into defined region(s) and therefore may become an important tool for evaluating the biological activity of potential drugs because drug activity and cell motility often directly correlate. PMID- 17129061 TI - Microemulsion-controlled reaction sites in biocatalytic films for electrochemical reduction of vicinal dibromides. AB - We report herein the electrochemical dehalogenation of vicinal dibromides in microemulsions using cross-linked films of the redox protein myoglobin (Mb) and poly-l-lysine (PLL) covalently bonded to carbon electrodes. Catalytic reduction of the dibromides to olefins was more efficient in an SDS microemulsion than in a CTAB microemulsion. SDS shifts the Mb redox potential more negative, but a comparison to Mb-SDS films suggests that the activation free energy of the reduction is controlled by an inner-sphere mechanism. SDS also enters the positively charged Mb-PLL films and preconcentrates the dibromide reactants, enhancing catalytic efficiency in SDS microemulsions. Shifts in formal potential and Soret absorbance bands for Mb-PLL films suggested binding of trans-1,2 dibromocyclohexane in the iron heme distal pocket with little catalysis. Results are consistent with active catalytic reduction sites for reactant bound on the protein surface and less-reactive sites in the distal heme pocket. Preconcentration into catalytic PLL films using SDS incorporated from microemulsions may be a general way to improve catalytic efficiency for nonpolar reactants in microemulsions. PMID- 17129062 TI - Comparison of the signaling and stability of electrochemical DNA sensors fabricated from 6- or 11-carbon self-assembled monolayers. AB - We have characterized the solution-phase and dry storage stability of electrochemical E-DNA sensors fabricated using mixed self-assembled monolayers (SAMs) composed of 6- or 11-carbon (C6 and C11, respectively) alpha,omega-thiol alcohols and the analogous C6- or C11-thiol-terminated stem-loop DNA probe. We find that the solution-phase and dry storage stability of C6-based E-DNA sensors are limited and poorly reproducible. The use of stabilizing agents bovine serum albumin plus either glucose or trehalose significantly improves the dry storage shelf life of such sensors: when using these preservatives, we observe only 7-9% sensor degradation after 1 month of storage in air at room temperature. In comparison, the stability of C11-based E-DNA sensors is significantly greater than that of the C6-based sensors; we observe only minor (5-8%) loss of signal upon storing these sensors for a week under ambient solution conditions or for more than a month in air in the presence of preservatives. Moreover, whereas the electron-transfer rate through C11 SAMs is slower than that observed for C6 SAMs, it is rapid enough to support good sensor performance. It thus appears that C11 SAMs provide a reasonable compromise between electron-transfer efficiency and sensor stability and are well suited for use in electronic DNA-sensing applications. PMID- 17129063 TI - Direct electron transfer--a favorite electron route for cellobiose dehydrogenase (CDH) from Trametes villosa. Comparison with CDH from Phanerochaete chrysosporium. AB - This paper presents some functional differences as well as similarities observed when comparing the newly discovered cellobiose dehydrogenase (CDH) from Trametes villosa (T.v.) with the well-characterized one from Phanerochaete chrysosporium (P.c.). The enzymes were physically adsorbed on spectrographic graphite electrodes placed in an amperometric flow through cell connected to a flow system. In the case of T.v.-CDH-modified graphite electrodes, a high direct electron transfer (DET) current was registered at the polarized electrode in the presence of the enzyme substrate reflecting a very efficient internal electron transfer (IET) process between the reduced FAD-cofactor and the oxidized heme cofactor. In the case of P.c.-CDH-modified graphite electrodes, the DET process is not as efficient, and the current will greatly increase in the presence of a mediator (mediated electron transfer, MET). As a consequence, when comparing the two types of enzyme-modified electrodes an inverted DET/MET ratio for T.v.-CDH is shown, in comparison with P.c.-CDH. The rates of the catalytic reaction were estimated to be comparable for both enzymes, by measuring the combined DET + MET currents. The inverted DET/MET ratio for T.v.-CDH-modified electrodes might suggest that probably there is a better docking between the two domains of this enzyme and that the linker region of P.c.-CDH might have an active role in modulating the rate of the IET (by changing the interdomain distance), with respect to pH. Based on the new properties of T.v.-CDH emphasized in the present study, an analytical application of a third-generation biosensor for lactose was recently published. PMID- 17129064 TI - Cyclic voltammetric responses of horseradish peroxidase multilayers on electrodes. AB - The catalytic responses obtained with step-by-step neutravidin-biotin deposition of successive monolayers of HRP are analyzed by means of cyclic voltammetry. The theoretical tools that have been developed allowed full characterization of the multilayered HRP coatings by means of a combination between closed-form analysis of limiting behaviors and finite difference numerical computations. An analysis of the experiments in which the number of monolayers was extended to 16 allowed an approximate determination of the average thickness of each monolayer, pointing to a compact arrangement of neutravidin and biotinylated HRP. The piling up of so many monolayers on the electrode allowed an improvement of the catalytic current by a factor of ca. 10, leading to very good sensitivities in term of cosubstrate detection. PMID- 17129065 TI - Electroactive self-assembled monolayers that permit orthogonal control over the adhesion of cells to patterned substrates. AB - This article describes an electroactive substrate that displays two independent dynamic functions for controlling the adhesion of cells. The approach is based on self-assembled monolayers on gold that are patterned into regions presenting the Arg-Gly-Asp peptide cell adhesion ligand. The patterned regions differ in the electrochemical properties of the linkers that tether the peptides to the monolayer. In this work, three distinct chemistries are employed that provide for release of the ligand on application of a negative potential, release of the ligand on application of a positive potential, and no change in response to a potential. Cells were allowed to attach to a monolayer patterned into circular regions comprising the three chemistries. Treatment with electric potentials of 650 or -650 mV resulted in the selective release of adherent cells only from regions that display the relevant electroactive groups. This example establishes the preparation of dynamic substrates with multiple functions and will be important to preparing model cultures derived from multiple cell types, with control over the temporal interactions of each cell population. PMID- 17129066 TI - Minor groove binding of a novel tetracationic diviologen. AB - Novel tetracationic diviologen compounds of the general formula CH3(CH2)nV2+(CH2)6V2+(CH2)nCH3 (where V2+ = 4,4'-bipyridinium and n = 5 or 11) were investigated as electrochemical reporters of DNA duplex formation. These compounds bind to both single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) when the DNA is either present in solution or immobilized at electrode surfaces. Binding to thiolated ssDNA and dsDNA immobilized at Au electrodes was characterized using the electrochemical response for the reduction of the V2+ state to the V+ (viologen radical cation) state. An analysis of the charge for this reduction provided isotherms and binding constants for binding of these diviologens to both forms of immobilized DNA. Saturation of the binding is achieved at solution concentrations near 20 microM. For both the n = 5 and 11 diviologens, binding to ssDNA is driven by electrostatic charge neutralization. For the n = 11 case, the binding is cooperative. In the presence of dsDNA, the n = 11 diviologen exhibits a unique reduction potential for the V2+/+ redox couple that is shifted approximately 100 mV negative of that in the presence of ssDNA. This new electrochemical signature is attributed to the reduction of viologen groups bound in the minor groove of the DNA duplex. For dsDNA in solution, an increase in the thermal denaturation temperature (Tm) from 60 to 66 degrees C as a function of the n = 11 diviologen concentration confirmed its interaction with the duplex. Circular dichroism (CD) spectroscopy also was used to investigate the binding of both the V2+ and V+ redox states of the n = 11 diviologen to dsDNA in solution. For the V+ state, a CD signal was observed that is consistent with the presence of face-to-face pi dimers of the viologen groups. This unambiguously demonstrates the binding of this redox state of the diviologen in the dsDNA minor groove and the formation of such dimers in the minor groove. PMID- 17129067 TI - S-nitrosothiol detection via amperometric nitric oxide sensor with surface modified hydrogel layer containing immobilized organoselenium catalyst. AB - A novel electrochemical device for the direct detection of S-nitrosothiol species (RSNO) is proposed by modifying an amperometric nitric oxide (NO) gas sensor with thin hydrogel layer containing an immobilized organoselenium catalyst. The diselenide, 3,3'-dipropionicdiselenide, is covalently coupled to primary amine groups in polyethylenimine (PEI), which is further cross-linked to form a hydrogel layer on a dialysis membrane support. Such a polymer film containing the organoselenium moiety is capable of decomposing S-nitrosothiols to generate NO(g) at the distal tip of the NO sensor. Under optimized conditions, various RSNOs (e.g., nitrosocysteine (CysNO), nitrosoglutathione (GSNO), etc.) are reversibly detected at 10-fold, respectively, indicating that the nature of the surfactant environment has a significant effect on the observed heme redox properties. PMID- 17129072 TI - Do pharmacogenomic tests provide value to policy makers? PMID- 17129073 TI - Letter from the commissioner's office at the US FDA. PMID- 17129074 TI - Assessing generalisability in model-based economic evaluation studies: a structured review in osteoporosis. AB - To support decision making, many countries have now introduced some formal assessment process to evaluate whether health technologies represent good 'value for money'. These often take the form of decision models that can be used to explore elements of importance to generalisability of study results across clinical settings and jurisdictions. The objective of this review was to assess whether articles reporting decision-analytic models in the area of osteoporosis provided enough information to enable decision makers in different countries/jurisdictions to fully appreciate the variability of results according to location and be able to apply the evaluation to their own setting. Of the 18 articles included in the review, only three explicitly stated the decision-making audience. It was not possible to infer a decision-making audience in eight studies. The target population was well reported, as were resource and cost data, and clinical data used for estimates of relative risk reduction. However, baseline risk was rarely adapted to the relevant jurisdiction, and when no decision maker was explicit it was difficult to assess whether the reported cost and resource use data were in fact relevant. A few studies used sensitivity analysis to explore elements of generalisability, such as compliance rates and baseline fracture risk rates, although such analyses were generally restricted to evaluating parameter uncertainty. This review found that variability in cost effectiveness across locations is addressed to a varying extent in modelling studies in the field of osteoporosis, limiting their use for decision makers across different locations. Transparency of reporting is expected to increase as methodology develops and decision makers publish 'reference case' type guidance. PMID- 17129075 TI - Generic health-related quality-of-life assessment in children and adolescents: methodological considerations. AB - The health-related quality of life (HR-QOL) of children and adolescents is increasingly considered a relevant topic for research. Instruments to assess quality of life in children and adolescents of a generic as well as disease- or condition-specific nature are being developed and applied in epidemiological surveys, clinical studies, quality assurance and health economics. This paper attempts to give an overview on the state of the art of HR-QOL assessment in children as it relates to methodological and conceptual challenges. Instruments available in international or cross-cultural research to assess HR-QOL in generic terms were identified and described according to psychometric data provided and the width of application. In an initial literature search, several challenges in the assessment of child and adolescent HR-QOL were identified, ranging from conceptual and methodological to practical aspects. Seven specific major issues were considered: (i) What are the dimensions of HR-QOL relevant for children and adolescents, and do suitable instruments for their measurement exist? (ii) Can these dimensions be collected in a cross-culturally comparable way? (iii) What advantages and disadvantages do self-rated versus externally evaluated HR-QOL measurements of children and adolescents have? (iv) How can HR-QOL be assessed in an age-appropriate way? (v) What are the advantages and disadvantages of disease specific and generic data collection? (vi) What advantages and disadvantages do profile and index instruments have? (vii) How can HR-QOL be connected with utility- preference values? In a second literature search we identified nine generic HR-QOL instruments and four utility health state classification systems that complied with the prespecified inclusion criteria. It was concluded that (i) HR-QOL instruments are available to assess the dimensions of the construct relevant to children and adolescents; (ii) provided that an instrument was constructed in an appropriate way, the dimensions of HR-QOL can be measured in an interculturally comparable manner; (iii) the HR-QOL of children and adolescents can and should be ascertained by self-rating; (iv) the measurement instruments used have to consider maturity and cognitive development; (v) only generic quality-of-life instruments allow for an assessment of HR-QOL in both healthy and chronically ill children and adolescents; (vi) the representation of HR-QOL achieved through a singular index value is connected to strict psychometric conditions: the index instrument has to be tailored to these psychometric conditions; (vii) how far utility measures are employable with children and adolescents has to be investigated in further studies. The problem aspects identified indicate the necessity for further research. Nevertheless, instruments for assessing the HR-QOL of children and adolescents can be identified that meet the requirements mentioned above. PMID- 17129076 TI - Cost effectiveness of tumour necrosis factor-alpha inhibitors as first-line agents in rheumatoid arthritis. AB - BACKGROUND AND OBJECTIVE: Rheumatoid arthritis (RA) is an autoimmune disease with an unknown aetiology that results in >9 million physician visits and >250 000 hospitalisations per year in the US. Tumour necrosis factor-alpha (TNFalpha) inhibitors are effective agents in treating RA; however, their cost effectiveness as first-line agents has not been investigated. This study aimed to examine the cost effectiveness of using TNFalpha inhibitors (both as monotherapy and in combination with methotrexate) as first-line agents versus methotrexate (monotherapy) from a payer perspective. METHODS: A Markov model was developed utilising a discount rate of 3% per annum, a cycle length of 1 year and a lifetime time-horizon for a hypothetical cohort of US females aged 55-60 years who had been diagnosed with RA. The source of data for predicted probabilities, expected mortality rates and treatment costs in year 2005 US dollars (drug, toxicity, monitoring and hospitalisation) was from the literature. These costs are assigned in 5-year cycles (calculated from initial 1-year estimates) along with the effect on quality-adjusted life-years (QALYs), which were calculated using the Health Assessment Questionnaire score. Univariate sensitivity analyses were conducted on all relevant parameters. RESULTS: Adalimumab, etanercept, adalimumab plus methotrexate and infliximab plus methotrexate had incremental cost-effectiveness ratios (ICERs) versus methotrexate monotherapy of $US63 769, $US89 772, $US194 589 and $US409 523 per QALY, respectively. When taking into consideration age at diagnosis, the ICER for etanercept ranged from $US84 129 to $US96 225 per QALY. In considering males for the base-case age at diagnosis, the ICER for etanercept versus methotrexate was $US85 100 per QALY. The average lifetime cost across all treatment arms in a woman diagnosed between 55 and 60 years of age was $US211 702. CONCLUSION: While these ICERs cannot be used to directly compare one biological agent with another since there are no comparative trials, they do provide a valid comparison versus methotrexate as first-line agents. Depending where the cost-effectiveness threshold is drawn (i.e. whether it is considered to be $US50 000 or $US100 000 per QALY), etanercept and adalimumab may be considered relatively cost-effective first-line treatments for RA compared with methotrexate monotherapy. PMID- 17129077 TI - Association of co-morbidities with prescribing patterns and cost savings: olanzapine versus risperidone for schizophrenia. AB - BACKGROUND: Olanzapine and risperidone are two commonly prescribed atypical antipsychotics for schizophrenia. Prior studies have shown inconsistent results in terms of advantage in cost saving in prescribing these agents. Our preliminary analysis showed that a small percentage of intensive healthcare utilizers had substantial impact on healthcare costs. This study analysed the cost effects of olanzapine and risperidone among those who had intensive utilisation of medical care prior to drug initiation, and the relationship between the choice of the two drugs and patients' co-morbid condition. METHODS: We retrospectively investigated patients first treated with either risperidone or olanzapine between 1 April 1999 and 31 March 2000. According to patients' medication history during the 6 months prior to initial prescription of a study medication we categorised patients into three groups: (i) not receiving olanzapine or risperidone; (ii) not receiving any atypical antipsychotic agent; or (iii) not receiving any antipsychotic agent. We then compared the difference in cost saving by type of care in the 10% most expensive patients through bivariate and multivariate analyses. Based on the records of 18 499 patients with schizophrenia prescribed either olanzapine or risperidone between 1 April 1999 and 31 March 2000 we defined intensive users of healthcare as those who incurred an annual total cost in the top tenth percentile. We measured co-morbidity by number of diseases, and healthcare costs ($US, 1998-2001 values) in the year prior and the year after treatment initiation in six categories of care (inpatient medical/surgical, inpatient psychiatric care, other inpatient, outpatient psychiatric care, other outpatient and outpatient pharmacy). RESULTS: The top 10% most expensive patients accounted for about half of the total cost of the entire cohort and had nearly a 40% cost reduction for the year after treatment initiation versus the prior year, while the entire cohort increased cost between 2% and 12%. Compared with those prescribed olanzapine, patients prescribed risperidone had more medical co morbidities, higher inpatient medical/surgical costs and lower psychiatric costs. Patients taking olanzapine had greater cost reduction in inpatient psychiatric care, whereas those taking risperidone had greater reduction in inpatient medical/surgical care. CONCLUSIONS: Among the top 10% most expensive patients, olanzapine and risperidone treatments were associated with comparable cost reductions in inpatient care. The choice of agent was associated with patients' co-morbid condition and was correlated with cost reduction in inpatient medical/surgical or psychiatric care. PMID- 17129079 TI - Discounting health effects in pharmacoeconomic evaluations: current controversies. PMID- 17129081 TI - Density functional theory reveals an increase in the amino 1H chemical shift in guanine due to hydrogen bonding with water. AB - Electronic structure calculations underestimate the chemical shift of the non H bonded amino proton 1H22 in isolated G-quartet structures. The current work shows that this underestimation is due to the absence of a water environment in the calculations: coordination of at least two water molecules is required to obtain good agreement with experiment. The results indicate how improved agreement between calculated and experimental (solution-phase) NMR data can be obtained. PMID- 17129078 TI - Candesartan cilexetil: a pharmacoeconomic review of its use in chronic heart failure and hypertension. AB - The addition of candesartan cilexetil (Atacand, Amias, Blopress, Kenzen, Ratacand) to standard therapy for chronic heart failure (CHF) provided important clinical benefits at little or no additional cost in France, Germany and the UK, according to a detailed economic analysis focusing on major cardiovascular events and prospectively collected resource-use data from the CHARM-Added and CHARM Alternative trials in patients with CHF and left ventricular (LV) systolic dysfunction. Results of a corresponding cost-effectiveness analysis showed that candesartan cilexetil was either dominant over placebo or was associated with small incremental costs per life-year gained, depending on the country and whether individual trial or pooled data were used. Preliminary data from a US cost-effectiveness analysis based on CHARM data also showed favourable results for candesartan cilexetil. Two cost-effectiveness analyses of candesartan cilexetil in hypertension have been published, both conducted in Sweden. Data from the SCOPE trial in elderly patients with hypertension, which showed a significant reduction in nonfatal stroke with candesartan cilexetil-based therapy versus non-candesartan cilexetil-based treatment, were incorporated into a Markov model and an incremental cost-effectiveness ratio of euro12 824 per QALY gained was calculated (2001 value). Another modelled cost-effectiveness analysis of candesartan cilexetil was based on the ALPINE trial, in which the incidence of new-onset diabetes was significantly lower in patients with newly diagnosed hypertension who were randomised to candesartan cilexetil (with or without felodipine) than among those who received hydrochlorothiazide (with or without atenolol). Although candesartan cilexetil was dominant over hydrochlorothiazide, the ALPINE cost-effectiveness analysis relied on a small number of clinical events and did not evaluate the incremental cost of candesartan cilexetil per life-year or QALY gained. In conclusion, despite some inherent limitations, economic analyses incorporating CHARM data and conducted primarily in Europe have shown that candesartan cilexetil appears to be cost effective when added to standard CHF treatment in patients with CHF and compromised LV systolic function. The use of candesartan cilexetil as part of antihypertensive therapy in elderly patients with elevated blood pressure was also deemed to be cost effective in a Swedish analysis, primarily resulting from a reduced risk of nonfatal stroke (as shown in the SCOPE study); however, the generalisability of results to other contexts has not been established. Cost-effectiveness analyses comparing candesartan cilexetil with ACE inhibitors or other angiotensin receptor blockers in CHF or hypertension are lacking, and results reported for candesartan cilexetil in a Swedish economic analysis of ALPINE data focusing on outcomes for diabetes require confirmation and extension. PMID- 17129082 TI - Magic alkali-fullerene compound clusters of extreme thermal stability. AB - The thermal stability of free pure C60-, as well as C60-alkali, and -alkaline earth metal compound clusters is investigated. We find that small (C60)m-clusters (mpi(CO)* transition by approximately 0.2 eV and a redshift of the O1s(OH)-->pi(CO)* by approximately 0.6 eV were observed, indicative of strong hydrogen-bond formation within the clusters. The results have been compared with a recent theoretical calculation, which supports the conclusion that the formic-acid clusters consist of the most stable cyclic dimer andor trimer units. Specifically labeled formic acid-d, HCOOD, was also used to examine the core-excited fragmentation mechanisms. These deuterium-labeled experiments showed that HDO+ was formed via site-specific migration of a formyl hydrogen within an individual molecule, and that HD2O+ was produced via the subsequent transfer of a deuterium atom from the hydroxyl group of a nearest-neighbor molecule within a cationic cluster. Deuteron (proton) transfer from the hydroxyl site of a hydrogen-bond partner was also found to take place, producing deuteronated HCOOD.D+ (protonated HCOOH.H+) cations within the clusters. PMID- 17129104 TI - The vibrational progressions of the N-->V electronic transition of ethylene: a test case for the computation of Franck-Condon factors of highly flexible photoexcited molecules. AB - The vibrational progressions of the N-->V electronic transition of ethylene--a test case for the computation of Franck-Condon factors between electronic states exhibiting very different equilibrium geometries--have been calculated by using both the Cartesian and the curvilinear internal coordinate representations of the normal modes of vibration. The comparison of the theoretical spectra with the experimental one shows that the Cartesian representation yields vibrational progressions which are not observed in the experimental spectrum, whereas the curvilinear one gives a very satisfying agreement, even in harmonic approximation. PMID- 17129105 TI - Role of rotational temperature in adiabatic molecular alignment. AB - One-dimensional alignment of molecules in the adiabatic limit, where the pulse duration greatly exceeds the molecular rotational periods, is studied experimentally. Four different asymmetric top molecules (iodobenzene, p diiodobenzene, 3,4-dibromothiophene, and 4,4'-dibromobiphenyl), rotationally cooled through a high pressure supersonic pulsed valve, are aligned by a 9-ns long pulse. Their orientations are measured through Coulomb explosion, induced by a 130-fs-long pulse, and by recording the direction of the recoiling ions. The paper focuses on the crucial role of the initial rotational temperature for the degree of alignment. In particular, we show that at molecular temperatures in the 1 K range very strong alignment is obtained already at intensities of a few times 10(11) W/cm2 for all four molecules. At the highest intensities (approximately 10(12) W/cm2) the molecules can tolerate without ionizing >or=0.92 in the case of iodobenzene. This is the strongest degree of alignment ever reported for any molecule. PMID- 17129106 TI - The electron affinity of phenanthrene. AB - Phenanthrene is studied by photodetachment-photoelectron spectroscopy. Due to the absence of a parent ion peak in the anion mass spectrum the electron affinity could not be determined directly. However, this absence is the first indication that this molecule has a negative electron affinity. The first three water complexes of phenanthrene were studied, supplying insights into its microsolvation property. Moreover, the electron affinity of the bare molecule could be determined to be -0.01+/-0.04 eV by an extrapolation method using the water cluster data. The experimental work is supported by ab initio calculations for determining the structure of the water complexes. Finally a correlation between the electron affinity and the reduction potential of polycyclic aromatic hydrocarbons is investigated. PMID- 17129107 TI - Manipulating spin-dependent interactions in rotationally excited cold molecules with electric fields. AB - We use rigorous quantum mechanical theory to study collisions of magnetically oriented cold molecules in the presence of superimposed electric and magnetic fields. It is shown that electric fields suppress the spin-rotation interaction in rotationally excited 2Sigma molecules and inhibit rotationally elastic and inelastic transitions accompanied by electron spin reorientation. We demonstrate that electric fields enhance collisional spin relaxation in 3Sigma molecules and discuss the mechanisms for electric field control of spin-changing transitions in collisions of rotationally excited CaD(2Sigma) and ND(3Sigma) molecules with helium atoms. The propensities for spin depolarization in the rotationally excited molecules are analyzed based on the calculations of collision rate constants at T=0.5 K. PMID- 17129108 TI - Dissociative and associative attachment of NO to iron clusters. AB - Electronic and geometrical structures of iron clusters with associative (FeNO, Fe2NO, Fe3NO, Fe4NO, Fe5NO, and Fe6NO) and dissociative (OFeN, OFe2N, OFe3N, OFe4N, OFe5N, and OFe6N) attachments of NO, as well as the corresponding singly negatively and positively charged ions, are computed using density functional theory with generalized gradient corrections. Both types of isomers are found to be stable and no spontaneous dissociation was observed during the geometry optimizations. The ground states correspond to dissociative attachment of NO for all iron clusters Fe(n), except for Fe and Fe+. All of the OFe(n)N clusters have ferrimagnetic ground states, except for OFe2N, OFe2N-, OFe4N, and OFe4N-, which prefer the ferromagnetic coupling. In the ferrimagnetic states, the excess spin density at one iron atom couples antiferromagnetically to the excess spin densities of all other iron atoms. Relative to the high-spin Fe(n) ground state, the lowest energy ferrimagnetic state quenches the total magnetic moments of iron clusters by 7, which is to be compared with a reduction in the magnetic moment of one in the lowest energy ferromagnetic states. Dissociation of NO on the iron clusters has a pronounced impact on the energetics of reactions; the Fe(n)NO+CO- >Fe(n)N+CO2 channels are exothermic while the OFe6N+CO--> Fe6N+CO2 channels are nearly thermoneutral. PMID- 17129109 TI - Rovibrational resonance effects in collision-induced electronic energy transfer: I2(E,v=0-2)+CF4. AB - Collisions of I2 in the E(0(g)+) electronic state with CF4 molecules induce electronic energy transfer to the nearby D, beta, and D' ion-pair states. Simulations of dispersed fluorescence spectra reveal collision-induced electronic energy transfer rate constants and final vibrational state distributions within each final electronic state. In comparison with earlier reports on I2(upsilon(E)=0-2) collisions with He or Ar atoms, we find markedly different dynamics when I2, excited to the same rovibronic states, collides with CF4. Final vibrational state distributions agree with the associated Franck-Condon factors with the initially prepared state to a greater degree than those found with He or Ar collision partners and suggest that internal degrees of freedom in the CF4 molecule represent a substantial means for accepting the accompanying loss of I2 vibronic energy. Comparison of the E-->D transfer of I2 excited to the J=23 and J=55 levels of the upsilon(E)=0 state reveals the onset of specific, nonstatistical dynamics as the available energy is increased above the threshold for excitation of the low frequency nu2 bending mode of CF4. PMID- 17129110 TI - C4Cl: bent or linear? AB - The ground state structure for the CCCCCl radical was computed by using symmetry adapted cluster configuration-interaction (SAC-CI) theory along with density functional theory to overcome the differences raised in the recently published paper [Y. Sumiyoshi et al., Chem. Phys. Lett. 414, 82 (2005)] between the theory and the experiment. SAC-CI results clearly support the earlier experimental conclusion that the radical has the bent ground state structure corresponding to 2Pi symmetry. Contrarily, probably due to spin contamination, mixing of a bent doublet ground state with the quartet components of a linear structure, coupled cluster singles and doubles (CCSD) calculations were unable to provide reliable results. Results obtained using density functional theory also show that the radical has a bent structure. Some low-lying doublet excited states were also studied using the SAC-CI theory. The energy difference between the ground Pi state and the nearby Sigma state is around 0.2 eV. The excitation energy for the transition with the largest oscillator strength agrees with the strongest absorption peak. PMID- 17129111 TI - Resonant two-photon ionization spectroscopy of BNB. AB - Triatomic BNB has been produced by laser ablation of a boron nitride rod in a supersonic expansion of helium carrier gas and has been investigated using resonant two-photon ionization spectroscopy in the visible region. The B 2Pi(g)-X 2Sigma(u)+ band system has been recorded near 514 nm and is dominated by a strong origin band, which has been rotationally resolved and analyzed. Both the (11)B(14)N(11)B (64% natural abundance) and the (10)B(14)N(11)B (32% natural abundance) isotopic modifications have been analyzed, leading to the spectroscopic constants (and their 1sigma error limits) of B0"(X 2Sigma(u)+)=0.466 147(70), B0'(B 2Pi(g))=0.467 255(75), and A0'(B 2Pi(g))=6.1563(38) cm(-1) for (10)B(14)N(11)B, corresponding to r(B-N)"(X 2Sigma(u)+)=1.312 47(10) A and r(B-N)'(B 2Pi(g))=1.310 92(11) A. Very similar values are obtained for the more abundant isotopomer, (11)B(14)N(11)B: B0"(X 2Sigma(u)+)=0.444 493(69), B0'(B 2Pi(g))=0.445 606(70), A0'(B 2Pi(g))=6.1455(38) cm(-1), corresponding to r(B-N)"(X 2Sigma(u)+)=1.312 41(10) A and r(B-N)'(B 2Pi(g))=1.310 77(10) A. These results are discussed as they relate to Walsh's rules and are compared to results for related molecules. PMID- 17129112 TI - A quantum-classical approach to the molecular dynamics of pyrazine with a realistic model Hamiltonian. AB - We investigate the molecular dynamics of pyrazine after excitation to the S2 electronic state by using the time-dependent discrete variable representation (TDDVR) method. The investigation has been carried out with a realistic 24-mode model Hamiltonian consisting of all the vibrational degrees of freedom of pyrazine molecule. First, we perform the simulation on a basic four-mode model, and then by including additional eight important modes and finally, by introducing 20 bath modes on the basic model. This sequential inclusion of bath modes demonstrates the effect of weak modes on the subsystem, where the calculations of energy and population transfer from basic model to the bath quantify the same effect. The spectral profile obtained by using TDDVR approach shows reasonably good agreement with the results calculated by quantum mechanical approach. It appears that the TDDVR approach for those large systems where quantum mechanical description is needed in a restricted region is a good compromise between accuracy and speed. PMID- 17129113 TI - Polarizabilities of the alkali anions: Li- to Fr-. AB - Static dipole polarizabilities are calculated for the ground states of the alkali anions from Li- to Fr-. The polarizabilities include scalar relativistic effects at the second-order Douglas-Kroll level and were computed using the finite-field, coupled-cluster CCSD(T) method with large, carefully optimized basis sets. The relativistic polarizabilities increase with Z, reach a maximum at Cs-, and then decrease again unlike their nonrelativistic counterparts which increase monotonically with Z. PMID- 17129114 TI - An experimental and theoretical study of double photoionization of CF4 using time of-flight photoelectron-photoelectron (photoion-photoion) coincidence spectroscopy. AB - Single photon double ionization of CF4 has been studied by means of a time-of flight photoelectron-photoelectron coincidence technique, which has very recently been extended towards ion detection, with energy analysis for the electrons and mass analysis for the ions. The complete single photon double ionization electron spectrum of CF4 up to a binding energy of approximately 51 eV is presented and discussed, also with the aid of accurate ab initio Green's function calculations. From ion detection in coincidence with the ejected electrons, we derive fragmentation pathway-selected double ionization electron spectra of CF4. From the same data we extract the yield of each doubly charged ion or ion pair as a function of the double ionization energy. PMID- 17129115 TI - Mode selective tunneling dynamics observed by high resolution spectroscopy of the bending fundamentals of 14NH2D and 14ND2H. AB - High resolution (0.004 and 0.01 cm(-1) instrumental bandwidth) interferometric Fourier transform infrared spectra of (14)NH2D and (14)ND2H were measured on a Bomem DA002 spectrometer in a supersonic jet expansion and at room temperature. We report the analysis of the bending fundamentals of (14)NH2D with term values Tv(s)=1389.9063(2) cm(-1) and Tv(a)=1390.4953(2) cm(-1) for the nu(4b) fundamental and Tv(s)=1605.6404(7) cm(-1) and Tv(a)=1591.0019(7) cm(-1) for the nu(4a) fundamental, and of (14)ND2H with term values of Tv(s)=1233.3740(2) cm(-1) and Tv(a)=1235.8904(2) cm(-1) for the nu(4a) fundamental and Tv(s)=1461.7941(9) cm(-1) and Tv(a)=1461.9918(19) cm(-1) for the nu(4b) fundamental. In all cases Tv(s) gives the position of the symmetric inversion sublevel (with positive parity) and Tv(a) the position of the antisymmetric inversion sublevel (with negative parity). The notation for the fundamentals nu(4a) and nu(4b) is chosen by correlation with the degenerate nu(4) mode in the C(3v) symmetric molecules NH3 and ND3. The degeneracy is lifted in Cs symmetry and a indicates the symmetric, b the antisymmetric normal mode with respect to the Cs symmetry plane in NH2D and ND2H. Assignments were established with certainty by means of ground state combination differences. About 20 molecular parameters of the effective S reduced Hamiltonian could be determined accurately for each fundamental. In particular, the effect of Fermi resonances of the 2nu(2) overtone with the nu(4a) bending mode was observed, leading to an increased inversion splitting in the case of ND2H and to a strongly increased inversion splitting and an inverted order of the two inversion levels in NH2D. Rotational perturbations observed with the nu(4b) bending fundamentals are probably due to Coriolis interactions with the inversion overtone 2nu(2). The results are important for understanding isotope effects on the inversion in ammonia as well as its selective catalysis and inhibition by excitation of different vibrational modes, as treated by quantum dynamics on high dimensional potential hypersurfaces of this molecule. PMID- 17129116 TI - C60 in intense short pulse laser fields down to 9 fs: excitation on time scales below e-e and e-phonon coupling. AB - The interaction of C60 fullerenes with 765-797 nm laser pulses as short as 9 fs at intensities of up to 3.7 x 10(14) W cm(-2) is investigated with photoion spectroscopy. The excitation time thus addressed lies well below the characteristic time scales for electron-electron and electron-phonon couplings. Thus, energy deposition into the system is separated from energy redistribution among the various electronic and nuclear degrees of freedom. Insight into fundamental photoinduced processes such as ionization and fragmentation is obtained from the analysis of the resulting mass spectra as a function of pulse duration, laser intensity, and time delay between pump and probe pulses, the latter revealing a memory effect for storing electronic energy in the system with a relaxation time of about 50 fs. Saturation intensities and relative abundances of (multiply charged) parent and fragment ions (C60(q+), q=1-6) are fingerprints for the ionization and fragmentation mechanisms. The observations indicate that for final charge states q>1 the well known C60 giant plasmon resonance is involved in creating ions and a significant amount of large fragments even with 9 fs pulses through a nonadiabatic multielectron dynamics. In contrast, for energetic reasons singly charged ions are generated by an essentially adiabatic single active electron mechanism and negligible fragmentation is found when 9 fs pulses are used. These findings promise to unravel a long standing puzzle in understanding C60 mass spectra generated by intense femtosecond laser pulses. PMID- 17129117 TI - Escape from a cavity through a small window: turnover of the rate as a function of friction constant. AB - To escape from a cavity through a small window the particle has to overcome a high entropy barrier to find the exit. As a consequence, its survival probability in the cavity decays as a single exponential and is characterized by the only parameter, the rate constant. We use simulations to study escape of Langevin particles from a cubic cavity through a small round window in the center of one of the cavity walls with the goal of analyzing the friction dependence of the escape rate. We find that the rate constant shows the turnover behavior as a function of the friction constant, zeta: The rate constant grows at very small zeta, reaches a maximum value which is given by the transition-state theory (TST), and then decreases approaching zero as zeta-->infinity. Based on the results found in simulations and some general arguments we suggest a formula for the rate constant that predicts a turnover of the escape rate for ergodic cavities in which collisions of the particle with the cavity walls are defocusing. At intermediate-to-high friction the formula describes transition between two known results for the rate constant: the TST estimation and the high friction limiting behavior that characterizes escape of diffusing particles. In this range of friction the rate constants predicted by the formula are in good agreement with those found in simulations. At very low friction the rate constants found in simulations are noticeably smaller than those predicted by the formula. This happens because the simulations were run in the cubic cavity which is not ergodic. PMID- 17129118 TI - The local and intermediate range structures of the five amorphous ices at 80 K and ambient pressure: a Faber-Ziman and Bhatia-Thornton analysis. AB - Using isotope substitution neutron scattering data, we present a detailed structural analysis of the short and intermediate range structures of the five known forms of amorphous ice. Two of the lower density forms--amorphous solid water and hyperquenched glassy water--have a structure very similar to each other and to low density amorphous ice, a structure which closely resembles a disordered, tetrahedrally coordinated, fully hydrogen bonded network. High density and very high density amorphous ices retain this tetrahedral organization at short range, but show significant differences beyond about 3.1 A from a typical water oxygen. The first diffraction peak in all structures is seen to be solely a function of the intermolecular organization. The short range connectivity in the two higher density forms is more homogeneous, while the hydrogen site disorder in these forms is greater. The low Q behavior of the structure factors indicates no significant density or concentration fluctuations over the length scale probed. We conclude that these three latter forms of ice are structurally distinct. Finally, the x-ray structure factors for all five amorphous systems are calculated for comparison with other studies. PMID- 17129119 TI - A new procedure for analyzing the nucleation kinetics of freezing in computer simulation. AB - A new method for deriving the size of the critical nucleus and the Zeldovich factor directly from kinetic data is presented. Moreover, in principle, the form of G(n), the free energy of formation of nuclei consisting of n molecules, can be inferred. The method involves measuring times of first appearance of nuclei of size n in the transient regime and applying the Becker-Doring theory. Times of first appearance exhibit the same characteristics as the conventional times associated with N(n,t), the number of nuclei of at least size n per unit volume that have materialized at time t. That is, they are well represented by three nucleation parameters, the reduced moment, the time lag, and the steady state nucleation rate. But unlike the conventional steady state rate which is independent of n, the steady state times of first appearance vary with n. In order to characterize the three nucleation parameters with precision, however, thousands of independent stochastic events with known n are required. Such sets of data are readily generated in molecular dynamic simulations but, so far, not in laboratory experiments. Results are illustrated by an analysis of simulations of the spontaneous freezing of large clusters of SeF6. PMID- 17129120 TI - Enzymatic catalysis and transfers in solution. I. Theory and computations, a unified view. AB - The transfer of hydride, proton, or H atom between substrate and cofactor in enzymes has been extensively studied for many systems, both experimentally and computationally. A simple equation for the reaction rate, an analog of an equation obtained earlier for electron transfer rates, is obtained, but now containing an approximate analytic expression for the bond rupture-bond forming feature of these H transfers. A "symmetrization," of the potential energy surfaces is again introduced [R. A. Marcus, J. Chem. Phys. 43, 679 (1965); J. Phys. Chem. 72, 891 (1968)], together with Gaussian fluctuations of the remaining coordinates of the enzyme and solution needed for reaching the transition state. Combining the two expressions for the changes in the difference of the two bond lengths of the substrate-cofactor subsystem and in the fluctuation coordinates of the protein leading to the transition state, an expression is obtained for the free energy barrier. To this end a two-dimensional reaction space (m,n) is used that contains the relative coordinates of the H in the reactants, the heavy atoms to which it is bonded, and the protein/solution reorganization coordinate, all leading to the transition state. The resulting expression may serve to characterize in terms of specific parameters (two "reorganization" terms, thermodynamics, and work terms), experimental and computational data for different enzymes, and different cofactor-substrate systems. A related characterization was used for electron transfers. To isolate these factors from nuclear tunneling, when the H-tunneling effect is large, use of deuterium and tritium transfers is of course helpful, although tunneling has frequently and understandably dominated the discussions. A functional form is suggested for the dependence of the deuterium kinetic isotope effect (KIE) on DeltaG degrees and a different form for the 13C KIE. Pressure effects on deuterium and 13C KIEs are also discussed. Although formulated for a one-step transfer of a light particle in an enzyme, the results would also apply to single-step transfers of other atoms and groups in enzymes and in solution. PMID- 17129121 TI - Pump-probe and pump-deplete-probe spectroscopies on carotenoids with N=9-15 conjugated bonds. AB - A series of all-trans-carotenoids with N=9, 13, and 15 conjugated bonds has been studied by pump-probe and pump-deplete-probe spectroscopies to obtain a systematic analysis of the energy flow between the different electronic states. The ultrafast dynamics in the carotenoids are initialized by excitation to the S2 state and subsequently manipulated by an additional depletion pulse in the near IR spectral range. The changes in the dynamics after depletion of the excited state population allowed differentiation of the excited state absorption into two components, a major one corresponding to the well known S1 state and the small contribution on the red wing of the S0-S2 absorption band originating from the hot ground state. We found no evidence for an additional electronically excited state, usually called S*. Instead, a deactivation mechanism that includes the hot ground state supports the observed results nicely in the framework of a simple three state model (S2, S1, and S0). PMID- 17129122 TI - Application of the optimized Baxter model to the hard-core attractive Yukawa system. AB - We perform Monte Carlo simulations on the hard-core attractive Yukawa system to test the optimized Baxter model that was introduced by Prinsen and Odijk [J. Chem. Phys. 121, 6525 (2004)] to study a fluid phase of spherical particles interacting through a short-range pair potential. We compare the chemical potentials and pressures from the simulations with analytical predictions from the optimized Baxter model. We show that the model is accurate to within 10% over a range of volume fractions from 0.1 to 0.4, interaction strengths up to three times the thermal energy, and interaction ranges from 6% to 20% of the particle diameter, and performs even better in most cases. We furthermore establish the consistency of the model by showing that the thermodynamic properties of the Yukawa fluid computed via simulations may be understood on the basis of one similarity variable, the stickiness parameter defined within the optimized Baxter model. Finally, we show that the optimized Baxter model works significantly better than an often used, naive method determining the stickiness parameter by equating the respective second virial coefficients based on the attractive Yukawa and Baxter potentials. PMID- 17129123 TI - Ab initio thermodynamics and phase diagram of solid magnesium: a comparison of the LDA and GGA. AB - The finite temperature density functional theory and quasiharmonic lattice dynamics have been used to compute numerous thermodynamic properties of hexagonal close packed magnesium using both the local density approximation (LDA) and the generalized gradient approximation (GGA) for the exchange-correlation potential. Generally, it is found that there exist only minor differences between the LDA and GGA computed properties, with both giving good agreement with experiment. The hcp-bcc phase boundary has also been computed and is found to be in agreement with experimental observation. Again, only slight differences are found between the LDA and GGA. PMID- 17129124 TI - Time dependent diffusion in a disordered medium with partially absorbing walls: a perturbative approach. AB - We present an analytical study of the time dependent diffusion coefficient in a dilute suspension of spheres with partially absorbing boundary condition. Following Kirkpatrick [J. Chem. Phys. 76, 4255 (1982)] we obtain a perturbative expansion for the time dependent particle density using volume fraction f of spheres as an expansion parameter. The exact single particle t operator for partially absorbing boundary condition is used to obtain a closed form time dependent diffusion coefficient D(t) accurate to first order in the volume fraction f. Short and long time limits of D(t) are checked against the known short time results for partially or fully absorbing boundary conditions and long time results for reflecting boundary conditions. For fully absorbing boundary condition the long time diffusion coefficient is found to be D(t)=5a(2)/(12fD(0)t)+O((D(0)t/a(2))(-2)) to the first order of perturbation theory. Here f is small but nonzero, D(0) the diffusion coefficient in the absence of spheres, and a the radius of the spheres. The validity of this perturbative result is discussed. PMID- 17129125 TI - Moving solvated electrons with light: nonadiabatic mixed quantum/classical molecular dynamics simulations of the relocalization of photoexcited solvated electrons in tetrahydrofuran (THF). AB - Motivated by recent ultrafast spectroscopic experiments [Martini et al., Science 293, 462 (2001)], which suggest that photoexcited solvated electrons in tetrahydrofuran (THF) can relocalize (that is, return to equilibrium in solvent cavities far from where they started), we performed a series of nonequilibrium, nonadiabatic, mixed quantum/classical molecular dynamics simulations that mimic one-photon excitation of the THF-solvated electron. We find that as photoexcited THF-solvated electrons relax to their ground states either by continuous mixing from the excited state or via nonadiabatic transitions, approximately 30% of them relocalize into cavities that can be over 1 nm away from where they originated, in close agreement with the experiments. A detailed investigation shows that the ability of excited THF-solvated electrons to undergo photoinduced relocalization stems from the existence of preexisting cavity traps that are an intrinsic part of the structure of liquid THF. This explains why solvated electrons can undergo photoinduced relocalization in solvents like THF but not in solvents like water, which lack the preexisting traps necessary to stabilize the excited electron in other places in the fluid. We also find that even when they do not ultimately relocalize, photoexcited solvated electrons in THF temporarily visit other sites in the fluid, explaining why the photoexcitation of THF-solvated electrons is so efficient at promoting recombination with nearby scavengers. Overall, our study shows that the defining characteristic of a liquid that permits the photoassisted relocalization of solvated electrons is the existence of nascent cavities that are attractive to an excess electron; we propose that other such liquids can be found from classical computer simulations or neutron diffraction experiments. PMID- 17129126 TI - Correlated hydrogen bonding fluctuations and vibrational cross peaks in N-methyl acetamide: simulation based on a complete electrostatic density functional theory map. AB - The coherent nonlinear response of the entire amide line shapes of N-methyl acetamide to three infrared pulses is simulated using an electrostatic density functional theory map. Positive and negative cross peaks contain signatures of correlations between the fundamentals and the combination state. The amide I-A and I-III cross-peak line shapes indicate positive correlation and anticorrelation of frequency fluctuations, respectively. These can be ascribed to correlated hydrogen bonding at C[double bond]O and N-H sites. The amide I frequency is negatively correlated with the hydrogen bond on carbonyl C[double bond]O, whereas the amide A and III are negatively and positively correlated, respectively, with the hydrogen bond on amide N-H. PMID- 17129127 TI - Glass transition in fullerenes: mode-coupling theory predictions. AB - We report idealized mode-coupling theory results for the glass transition of ensembles of model fullerenes interacting via phenomenological two-body potentials. Transition lines are found for C60, C70, and C96 in the temperature density plane. We argue that the observed glass transition behavior is indicative of kinetic arrest that is strongly driven by the interparticle attraction in addition to excluded-volume repulsion. In this respect, these systems differ from most standard glass-forming liquids. They feature arrest that occurs at lower densities and that is stronger than would be expected for repulsion-dominated hard-sphere-like or Lennard-Jones-type systems. The influence of attraction increases with increasing the number of carbon atoms per molecule. However, unrealistically large fullerenes would be needed to yield behavior reminiscent of recently investigated model colloids with strong short-ranged attraction (glass glass transitions and logarithmic decay of time-correlation functions). PMID- 17129128 TI - Cluster pair correlation function of simple fluids: energetic connectivity criteria. AB - We consider the clustering of Lennard-Jones particles by using an energetic connectivity criterion proposed long ago by Hill [J. Chem. Phys. 32, 617 (1955)] for the bond between pairs of particles. The criterion establishes that two particles are bonded (directly connected) if their relative kinetic energy is less than minus their relative potential energy. Thus, in general, it depends on the direction as well as on the magnitude of the velocities and positions of the particles. An integral equation for the pair connectedness function, proposed by two of the authors [Phys. Rev. E 61, R6067 (2000)], is solved for this criterion and the results are compared with those obtained from molecular dynamics simulations and from a connectedness Percus-Yevick-type integral equation for a velocity-averaged version of Hill's energetic criterion. PMID- 17129129 TI - Non-Markovian modification of the golden rule rate expression. AB - The reformulation of the standard golden rule approach considered in this paper for treating reactive tunneling reduces the computation of the reaction rate to a derivation of band shapes for energy levels of reactant and product states. This treatment is based on the assumption that the medium environment is actively involved as a partner in the energy exchange with the reactive subsystem but its reorganization effect is negligible. Starting from the quantum relaxation equation for the density matrix, the required band shapes are represented in terms of the spectral density function, exhibiting the continuum spectrum inherent to the interaction between the reactants and the medium in the total reactive system. The simplest Lorentzian spectral bands, obtained under Redfield approximation, proved to be unsatisfactory because they produced a divergent rate expression at low temperature. The problem is resolved by invoking a refined spectral band shape, which behaves as Lorentzian one at the band center but decays exponentially at its tails. The corresponding closed non-Markovian rate expression is derived and investigated taking as an example the photochemical H transfer reaction between fluorene and acridine proceeding in the fluorene molecular crystal. The kinetics in this reactive system was thoroughly studied experimentally in a wide temperature range [B. Prass et al., Ber. Bunsenges. Phys. Chem. 102, 498 (1998)]. PMID- 17129130 TI - The model of level broadening in condensed phase. AB - We study a model of non-Markovian kinetics for a harmonic oscillator embedded in a harmonic heat bath. We present a new scheme for approximately solving the quantum relaxation equation for the density matrix to find a distribution of level populations. It is found to be an extended Lorentzian with the width depending on the energy. A more convenient non-Markovian distribution called square root Fourier distribution that was implemented in the preceding paper [M. V. Basilevsky et al., J. Chem. Phys. 125, 194513 (2006)] is closely related to this extended Lorentzian model. Both distributions decay exponentially far away from their centers and reproduce well standard Lorentzian widths in the vicinity of the central region. A conventional Lorentzian model with such widths results when the Redfield approximation is applied in the frame of the present procedure. PMID- 17129131 TI - Designing an enzymatic oscillator: bistability and feedback controlled oscillations with glucose oxidase in a continuous flow stirred tank reactor. AB - The reaction of glucose with ferricyanide catalyzed by glucose oxidase from Aspergillus niger gives rise to a wide range of bistability as the flow rate is varied in a continuous flow stirred tank reactor. Oscillations in pH can be obtained by introducing a negative feedback on the autocatalytic production of H+ that drives the bistability. In our experiments, this feedback consists of an inflow of hydroxide ion at a rate that depends on [H+] in the reactor as k0[OH ]0[H+]/(K+[H+]). pH oscillations are found over a broad range of enzyme and ferricyanide concentrations, residence times (k0 (-1)), and feedback parameters. A simple mathematical model quantitatively accounts for the experimentally found oscillations. PMID- 17129132 TI - Study on the vibrational energy relaxation of p-nitroaniline, N,N-dimethyl-p nitroaniline, and azulene by the transient grating method. AB - The vibrational energy dissipation processes of the electronic ground states of p nitroaniline and N,N-dimethyl-p-nitroaniline have been studied by transient grating spectroscopy with subpicosecond laser pulses. The rise time of the acoustic signal produced by the energy dissipation process of the hot ground state molecule was monitored. The acoustic signal was analyzed by an equation including the acoustic damping. The solvent temperature rise times in various solvents have been determined. The acoustic signals of azulene in previous papers [Y. Kimura et al., J. Chem. Phys. 123, 054512 (2005); 123, 054513 (2005)] were also reanalyzed using this equation. The temperature rise times in all cases are longer than the vibrational energy relaxation times of the solutes determined by the transient absorption measurements. The difference is discussed in terms of the energy transfer pathways from the solute to the solvent. We concluded that both the hydrogen bonding between the solute and the solvent and the lower frequency modes of the solutes play important roles in determining the energy transfer pathway from the solute to the solvent. PMID- 17129133 TI - Adiabatic transfer of population in a dense fluid: the role of dephasing statistics. AB - We report the results of simulation studies of the statistics of vibrational dephasing of a YCl (Y=H, D, T, and I) diatom in dense fluid Ar at two temperatures, including the effect of strong field driving on the energy level modulation statistics. The distribution of energy level modulations is found to be non-Gaussian with a high energy tail. Aspects of stimulated Raman adiabatic passage (STIRAP) between the vibrational levels of HCl in dense fluid Ar have been investigated. For HCl with nearly degenerate v=0-->v=1 and v=1-->v=2 transitions, the combined effect of modulation and power broadening reduces the STIRAP efficiency for population transfer from v=0 to v=2 of the order of 30%. However, if the transitions used have very different frequencies, as in the original model studied by Demirplak and Rice [J. Chem. Phys. 116, 8028 (2002)], the STIRAP efficiency for population transfer remains high, of the order of 80%, even with non-Gaussian modulation of energy levels. PMID- 17129134 TI - Can the pH value of water solutions be estimated by quantum chemical calculations of small water clusters? AB - In the study, various water clusters were explored from the point of view of the proton transfer between H-bonded neighbors. A relatively modest approach--the MP26-31++G(d,p) level--was chosen as acceptable considering the fact that also larger systems have to be included. The tight ion-pair model (with usually three fixed O-O distances) was adopted for the autodissociation process. First, cluster estimated pH values rapidly decrease as cluster size increases from 2 to 6. For larger clusters in gas phase, the topology of H bonds plays an important role, varying pH from 7 to 13 in hexamers and from 5 to 15 in octamer clusters. The relationship energy/distance was quantified, too. Enhancing our model with the conductorlike screening model (COSMO) approach brought significant improvement in description of the autodissociation reaction with a stable zwitterionic structure. However, when the O-O restrictions were released, the small barrier for backward reaction disappeared, reforming neutral cluster spontaneously. Also Boltzmann weighting procedure was applied, and for the explored clusters in vacuo, the series of pH 25-18-14-13-10 was obtained for cluster sizes n=2, 3, 4, 6, 8. Using the COSMO approach, the analogous series is 15-14-12-10-9. The limit of the series is still about two to three units above the experimentally known pH. In order to reach the size-independent (bulk) value, larger clusters are needed. However, the situation is far from hopeless since (as it was proven in the study) four-coordinated molecules are not involved in the proton transfer process directly; they can only be a part of the surrounding environment. PMID- 17129135 TI - Alternative fundamental measure theory for additive hard sphere mixtures. AB - The purpose of this short paper is to present an alternative fundamental measure theory (FMT) for hard sphere mixtures. Keeping the main features of the original Rosenfeld's FMT [Phys. Rev. Lett. 63, 980 (1989)] and using the dimensional and the low-density limit conditions a new functional is derived incorporating Boublik's multicomponent extension [Mol. Phys. 59, 371 (1986)] of highly accurate Kolafa's equation of state for pure hard spheres. We test the theory for pure hard spheres and hard sphere mixtures near a planar hard wall and compare the results with the original Rosenfeld's FMT and one of its modifications and with new very accurate simulation data. The test reveals an excellent agreement between the results based on the alternative FMT and simulation data for density profile near a contact and some improvement over the original Rosenfeld's FMT and its modification at the contact region. PMID- 17129136 TI - Mixed quantum-classical description of spectroscopy of dissipative systems. AB - Mixed quantum-classical statistical mechanics is employed to calculate dipole moment correlation function and linear absorption spectra. A quantum two-level subsystem interacting with quantum vibrations (primary oscillators) which in turn are coupled to a classical bath composed of infinite set of harmonic oscillators is used as a dissipative system. Starting with mixed quantum-classical Liouville equation for the evaluation of the mixed quantum-classical dipole moment correlation function and using coherent states and the inverse of Baker-Campbell Hausdorf formula to evaluate the trace over the primary oscillators, whereby, a closed analytical expression for the electronic dipole moment correlation function is obtained. Illustrations of several absorption spectra at different temperatures are provided. An approximate optical four-point correlation is obtained in the high temperature limit. A strategy for deriving an exact optical four-point correlation is suggested. PMID- 17129137 TI - Multidimensional infrared spectroscopy of water. I. Vibrational dynamics in two dimensional IR line shapes. AB - In this and the following paper, we describe the ultrafast structural fluctuations and rearrangements of the hydrogen bonding network of water using two-dimensional (2D) infrared spectroscopy. 2D IR spectra covering all the relevant time scales of molecular dynamics of the hydrogen bonding network of water were studied for the OH stretching absorption of HOD in D2O. Time-dependent evolution of the 2D IR line shape serves as a spectroscopic observable that tracks how different hydrogen bonding environments interconvert while changes in spectral intensity result from vibrational relaxation and molecular reorientation of the OH dipole. For waiting times up to the vibrational lifetime of 700 fs, changes in the 2D line shape reflect the spectral evolution of OH oscillators induced by hydrogen bond dynamics. These dynamics, characterized through a set of 2D line shape analysis metrics, show a rapid 60 fs decay, an underdamped oscillation on a 130 fs time scale induced by hydrogen bond stretching, and a long time decay constant of 1.4 ps. 2D surfaces for waiting times larger than 700 fs are dominated by the effects of vibrational relaxation and the thermalization of this excess energy by the solvent bath. Our modeling based on fluctuations with Gaussian statistics is able to reproduce the changes in dispersed pump-probe and 2D IR spectra induced by these relaxation processes, but misses the asymmetry resulting from frequency-dependent spectral diffusion. The dynamical origin of this asymmetry is discussed in the companion paper. PMID- 17129138 TI - Multidimensional infrared spectroscopy of water. II. Hydrogen bond switching dynamics. AB - We use multidimensional infrared spectroscopy of the OH stretch of HOD in D2O to measure the interconversion of different hydrogen bonding environments. The OH stretching frequency distinguishes hydrogen bonded (HB) and non-hydrogen-bonded (NHB) configurations by their absorption on the low (red) and high (blue) sides of the line shape. Measured asymmetries in the two dimensional infrared OH line shapes are manifestations of the fundamentally different spectral relaxations of HB and NHB. HB oscillators exhibit coherent oscillations within the hydrogen bonded free energy well before undergoing activated barrier crossing, resulting in the exchange of hydrogen bonded partners. Conversely, NHB oscillators rapidly return to HB frequencies within 150 fs. These results support a picture where NHB configurations are only visited transiently during large fluctuations about a hydrogen bond or during the switching of hydrogen bonding partners. The results are not consistent with the presence of entropically stabilized dangling hydrogen bonds or a conceptual picture of water as a mixture of environments with varying hydrogen bond strength separated by barriers >kT. PMID- 17129139 TI - Effects of multivalent salt addition on effective charge of dilute colloidal solutions. AB - The effective charge Z* is often invoked to account for the accumulation of counterions near the colloid with intrinsic charge Z. Although the ion concentrations c(i) are not uniform in the solution due to the presence of the charged particle, their chemical potentials are uniform everywhere. Thus, on the basis of ion chemical potential, effective ion concentrations c(i)*, which can be experimentally measured by potentiometry, are defined with the pure salt solution as the reference state. The effective charge associated with the charged particle can then be determined by the global electroneutrality condition. Monte Carlo simulations are performed in a spherical Wigner-Seitz cell to obtain the effective charge of the colloid. In terms of the charge ratio alpha=Z*/Z, the effects of added salt concentration, counterion valency, and particle charge are examined. The effective charge declines with increasing salt concentration and the multivalent salt is much more efficient in reducing the effective charge of the colloidal solution. Moreover, the extent of effective charge reduction is decreased with increasing intrinsic charge for a given concentration of added salt. Those results are qualitatively consistent with experimental observations by electrophoresis. PMID- 17129140 TI - Infrared and Raman spectra of silica polymorphs from an ab initio parametrized polarizable force field. AB - The general aim of this study is to test the reliability of polarizable model potentials for the prediction of vibrational (infrared and Raman) spectra in highly anharmonic systems such as high temperature crystalline phases. By using an ab initio parametrized interatomic potential for SiO2 and molecular dynamics simulations, we calculate the infrared and Raman spectra for quartz, cristobalite, and stishovite at various thermodynamic conditions. The model is found to perform very well in the prediction of infrared spectra. Raman peak positions are also reproduced very well by the model; however, Raman intensities calculated by explicitly taking the derivative of the polarizability with respect to the atomic displacements are found to be in poorer agreement than intensities calculated using a parametrized "bond polarizability" model. Calculated spectra for the high temperature beta phases, where the role of dynamical disorder and anharmonicities is predominant, are found to be in excellent agreement with experiments. For the octahedral phases, our simulations are able to reproduce changes in the Raman spectra across the rutile-to-CaCl2 transition around 50 GPa, including the observed phonon softening. PMID- 17129141 TI - X-ray diffraction and inelastic neutron scattering study of 1:1 tetramethylpyrazine chloranilic acid complex: temperature, isotope, and pressure effects. AB - The x-ray diffraction studies of the title complex were carried out at room temperature and 14 K for H/D (in hydrogen bridge) isotopomers. At 82 K a phase transition takes place leading to a doubling of unit cells and alternation of the hydrogen bond lengths linking tetramethylpyrazine (TMP) and chloranilic acid molecules. A marked H/D isotope effect on these lengths was found at room temperature. The elongation is much smaller at 14 K. The infrared isotopic ratio for O-H(D)...N bands equals to 1.33. The four tunnel splittings of methyl librational ground states of the protonated complex required by the structure are determined at a temperature T=4.2 K up to pressures P=4.7 kbars by high resolution neutron spectroscopy. The tunnel mode at 20.6 microeV at ambient pressure shifts smoothly to 12.2 microeV at P=3.4 kbars. This is attributed to an increase of the strength of the rotational potential proportional to r(-5.6). The three other tunnel peaks show no or weak shifts only. The increasing interaction with diminishing intermolecular distances is assumed to be compensated by a charge transfer between the constituents of deltae/e approximately 0.02 kbar(-1). The phase transition observed between 3.4 and 4.7 kbars leads to increased symmetry with only two more intense tunneling bands. In the isotopomer with deuterated hydrogen bonds and P=1 bar all tunnel intensities become equal in consistency with the low temperature crystal structure. The effect of charge transfer is confirmed by a weakening of rotational potentials for those methyl groups whose tunnel splittings were independent of pressure. Density functional theory calculations for the model TMP.(HF)2 complex and fully ionized molecule TMP+ point out that the intramolecular rotational potential of methyl groups is weaker in the charged species. They do not allow for the unequivocal conclusions about the role of the intermolecular charge transfer effect on the torsional frequencies. PMID- 17129142 TI - A crossover from metal to plasma in dense fluid hydrogen. AB - Thermodynamic properties in dense fluid hydrogen are studied by using a density functional theory for electron-proton binary mixtures that is called quantal hypernetted-chain (QHNC) integral equation. A nonlocal approximation for the exchange-correlation potential in a finite-temperature Kohn-Sham equation is presented. Results obtained from the QHNC with the nonlocal approximation are compared with those obtained from the QHNC with a local density approximation. Temperature variation of thermodynamic quantities between 10(4) and 10(6) K are investigated along an isochor specified by a dimensionless density parameter of rs=0.5. These quantities obtained from the QHNCs show that a crossover from metal to plasma occurs around a temperature of T=1.78 x 10(5) K. Electrical resistivity Re of the dense fluid hydrogen evaluated from a Ziman formula [The Properties of Liquid Metals, edited by S. Takenohi (Wiley, New York, 1973)] extended to finite temperature is about 0.7 muOmega cm at T=10(4) K. The dense fluid hydrogen at the temperature can be considered as a metallic fluid, because the value is smaller than typical values of Re in alkali metals at room temperature. The Re slightly increases with the temperature increase, and the temperature valuation of Re is monotonic. We clearly show that the contribution from the electronic excited states plays an important role for the sharp crossover from the metal to the plasma, and that the crossover is interpreted as a crossover from degenerate electron gas to nondegenerate electron gas. PMID- 17129143 TI - Prediction of viscosity for molecular fluids at experimentally accessible shear rates using the transient time correlation function formalism. AB - Nonequilibrium molecular dynamics (NEMD) simulations were performed and the transient time correlation function (TTCF) method applied to calculate the shear viscosity of n-decane. Using the TTCF method we were able to calculate the viscosity at shear rate orders of magnitude lower than is possible by direct NEMD simulation alone. For the first time for a molecular fluid, we were able to simulate shear rates accessible by experimental measurements, which are typically performed at shear rates well below those accessible by NEMD simulation. The TTCF method allows us to close the gap between the lowest shear rates accessible by MD simulation and the highest shear rates possible in experimental studies. Additionally a multiple time step method for Gaussian thermostatted SLLOD equations of motion was developed following earlier work [G. A. Pan et al., J. Chem. Phys. 122, 4114 (2005)] for atomic fluids. PMID- 17129144 TI - Phase transitions in DNA-linked nanoparticle assemblies: a decorated-lattice model. AB - We use decorated-lattice models to explore the phase behavior of two types of DNA linked colloidal mixtures: systems with identical nanoparticles functionalized with two different DNA strands (mixture Aab) and mixtures involving two types of particles each one functionalized with a different DNA strand (mixture Aa-Ab). The model allows us to derive the properties of the mixtures from the well-known behavior of underlying spin-n Ising models with temperature and activity dependent effective interactions. The predicted evolution of the dissolution profiles for the colloidal assemblies as a function of temperature and number of single DNA strands on a nanoparticle M is in qualitative agreement with that observed in real systems. According to our model, the temperature at which the assemblies dissolve can be expected to increase with increasing M only for concentrations of colloids below a certain threshold. For more concentrated solutions, the dissolution temperature is a decreasing function of M. Linker mediated interactions between Aa and Ab particles in the Aa-Ab mixture render the phase separation involving disordered aggregates metastable with respect to a phase transition between a solvent-rich and an ordered phase. The stability of the DNA-linked assembly is enhanced by the ordering of the colloidal network and the ordered aggregates dissolve at higher temperatures. Our results may explain the contrasting evolution of the dissolution temperatures with increasing probe size in Aab and Aa-Ab mixtures as observed experimentally. PMID- 17129145 TI - Dynamic strength of molecularly bonded surfaces. AB - This study reports a theoretical analysis of the forced separation of two adhesive surfaces linked via a large number of parallel noncovalent bonds. To describe the bond kinetics, we implement a three-state reaction model with kinetic rates obtained from a simple integral expression of the mean first passage time for diffusive barrier crossing in a pulled-distance-dependent potential. We then compute the rupture force for the separation of adhesive surfaces at a constant rate. The results correspond well with a Brownian dynamics simulation of the same system. The separation rate relative to the intrinsic relaxation time of the bonds defines three loading regimes and the general dependence of the adhesion on kinetic or thermodynamic parameters of the bonds. In the equilibrium regime, the rupture force asymptotically approaches the equilibrium rupture force, which increases linearly with the equilibrium bond energy. In the near-equilibrium regime, the rupture force increases with the separation rate and increasingly correlates with the bond rupture barrier. In the far-from-equilibrium regime where rebinding is irrelevant, the rupture force varies linearly with the rupture barrier. PMID- 17129146 TI - Effects of hydrogen bonding on current-voltage characteristics of molecular junctions. AB - We present a first-principles study of hydrogen bonding effect on current-voltage characteristics of molecular junctions. Three model charge-transfer molecules, 2' amino-4,4'-di(ethynylphenyl)-1-benzenethiolate (DEPBT-D), 4,4'-di(ethynylphenyl) 2'-nitro-1-benzenethiolate (DEPBT-A), and 2'-amino-4,4'-di(ethynylphenyl)-5' nitro-1-benzenethiolate (DEPBT-DA), have been examined and compared with the corresponding hydrogen bonded complexes formed with different water molecules. Large differences in current-voltage characteristics are observed for DEPBT-D and DEPBT-A molecules with or without hydrogen bonded waters, while relatively small differences are found for DEPBT-DA. It is predicted that the presence of water clusters can drastically reduce the conductivities of the charge-transfer molecules. The underlying microscopic mechanism has been discussed. PMID- 17129147 TI - Rotatoelectricity in cholesteric side-chain liquid single crystal elastomers. AB - We analyze the phenomenon of rotatoelectricity which is characteristic of cholesteric side-chain liquid single crystal elastomers. Using a linearized macroscopic continuum description and our previous work we show that if such a material is exposed to a static external electric field oriented parallel to the cholesteric helical axis, the director of the liquid crystalline phase will rotate around the helical axis. The material considered is assumed to be a perfect electric insulator. We propose an experiment in which the effect of rotatoelectricity should be directly observable and from which the ratio of the material parameters involved will be accessible. PMID- 17129148 TI - Thiol and thiolate bond formation of ferrocene-1,1-dithiol to a Ag(111) surface. AB - Using density functional calculations, we show that the adsorption of ferrocene dithiol on the Ag(111) surface is remarkably flexible, i.e., a large number of different configurations have binding energies that differ by less than 0.1 eV per molecule. The thiolate bond is slightly favored over the thiol bond (by less than 0.1 eV) but may not be formed due to considerable activation barriers. Electronically, we found that the thiolate bound molecule is conducting, whereas thiol bonds turn it into semiconducting. PMID- 17129149 TI - The oxidized soot surface: theoretical study of desorption mechanisms involving oxygenated functionalities and comparison with temperature programed desorption experiments. AB - The desorption mechanism for oxygenated functionalities on soot is investigated by quantum mechanical calculations on functionalized polycyclic aromatic hydrocarbon (PAH) models and compared with recently published temperature programed desorption-mass spectrometry results. Substituents on PAHs of increasing size (up to 46 carbon atoms in the parent PAH) are chosen to reproduce the local features of an oxidized graphenic soot platelet. Initially, the study is carried out on unimolecular fragmentation (extrusion, in some cases) processes producing HO, CO, or CO2, in model ketones, carboxylic acids, lactones, anhydrides, in one aldehyde, one peroxyacid, one hydroperoxide, one secondary alcohol, and one phenol. Then, a bimolecular process is considered for one of the carboxylic acids. Furthermore, some cooperative effect which can take place by involving two vicinal carboxylic groups (derived from anhydride hydrolysis) is investigated for other four bifunctionalized models. The comparison between the computed fragmentation (desorption) barriers for the assessed mechanisms and the temperature at which maxima occur in TPD spectra (for HO, CO, or CO2 desorption) offers a suggestion for the assignment of these maxima to specific functional groups, i.e., a key to the description of the oxidized surface. Notably, the computations suggest that (1) the desorption mode from a portion of a graphenic platelet functionalized by a carboxylic or lactone groups is significantly dependent from the chemical and geometric local environment. Consequently, we propose that (2) not all carboxylic groups go lost at the relatively low temperatures generally stated, and (3) lactone groups can be identified as producing not only CO2 but also CO. PMID- 17129150 TI - CO adsorption on pure and binary-alloy gold clusters: a quantum chemical study. AB - We performed density-functional theory analysis of nondissociative CO adsorption on 22 binary Au-alloy (Au(n)M(m)) clusters: n=0-3, m=0-3, and m+n=2 (dimers) or 3 (trimers), M=Cu/Ag/Pd/Pt. We report basis-set superposition error corrections to adsorption energies and include both internal energy of adsorption (DeltaU(ads)) and Gibbs free energy of adsorption (DeltaG(ads)) at standard conditions (298.15 K and 1 atm). We found onefold (atop) CO binding on all the clusters except Pd2 (twofold/bridged), Pt2 (twofold/bridged), and Pd3 (threefold). In agreement with the experimental results, we found that CO adsorption is thermodynamically favorable on pure Au/Cu clusters but not on pure Ag clusters and also observed the following adsorption affinity trend: Pd>Pt>Au>Cu>Ag. For alloy dimers we found the following patterns: Au2>M Au>M2 (M=Ag/Cu) and M2>M Au>Au2 (M=Pd/Pt). Alloying Ag/Cu dimers with (more reactive) Au enhanced adsorption and the opposite effect was observed for PdPt dimers. The Ag-Au, Cu-Au, and Pd-Au trimers followed the trends observed on dimers: Au3>M Au2>M2Au>M3 (M=Ag/Cu) and Pd3>Pd2Au>PdAu2>Au3. Interestingly, Pt-Au trimers reacted differently and alloying with Au systematically increased the adsorption affinity: PtAu2>Pt2Au>Pt3>Au3. A strikingly different behavior of Pt is also manifested by the triplet spin state and onefold (atop) binding in Pt3-CO which is in contradiction with the singlet spin state and threefold binding in Pd3-CO. We found a linear correlation between CO binding energy (BE) and elongation of the CO bond. For Ag-Au and Cu-Au clusters, the increase in CO BE (and elongation of the C-O bond which is probably due to the back donation) is accompanied by the decrease in the cluster-CO distance suggesting that the donation (from 5sigma highest occupied molecular orbital in CO to cluster lowest unoccupied molecular orbital) mechanism also contributes to the BE. For Pd-Au clusters, the cluster-CO distance (and CO bond length) increases with increase in the BE, suggesting that the donation mechanism may not be important for those clusters. No clear trend was observed for Pt-Au clusters. PMID- 17129151 TI - Competition between double exchange and purely magnetic Heisenberg models in mixed valence systems: application to half-doped manganites. AB - A truncated Hubbard model is developed for the description of the electronic structure of odd-electron TM-L-TM units (TM=transition metal and L=ligand). The model variationally treats both the double exchange and purely magnetic Heisenberg configurations. This Hubbard model can either be mapped on a purely magnetic Heisenber model in which the bridging oxygen is also magnetic or on a double exchange model owing to the hybridization of the magnetic and ligand or bitals. The purely magnetic Heisenberg model is analytically solved in the general case of two metals (having n magnetic orbitals) bridged by a magnetic oxygen. The comparison of the analytical expressions of the Heisenberg energies to those of the double exchange model reveals that the two model spectra are identical except for one state which does not belong to the model space of the double exchange Hamiltonian. Consequently, the fitting of the model spectra to accurate ab initio spectra does not discriminate between the physically different models. These concepts are illustrated for the Mn-O-Mn unit (or Zener polaron) found in the half-doped manganite Pr(0.6)Ca(0.4)MnO3. It is shown that in the present case the projections of the ab initio ground state wave function onto both model spaces are almost identical provided that one uses properly localized orbitals, proving that the magnetic description of the Zener polaron and the double exchange viewpoint of the electronic structure are equally valid. PMID- 17129152 TI - Geometrical cluster ensemble analysis of random sphere packings. AB - We introduce a geometric analysis of random sphere packings based on the ensemble averaging of hard-sphere clusters generated via local rules including a nonoverlap constraint for hard spheres. Our cluster ensemble analysis matches well with computer simulations and experimental data on random hard-sphere packing with respect to volume fractions and radial distribution functions. To model loose as well as dense sphere packings various ensemble averages are investigated, obtained by varying the generation rules for clusters. Essential findings are a lower bound on volume fraction for random loose packing that is surprisingly close to the freezing volume fraction for hard spheres and, for random close packing, the observation of an unexpected split peak in the distribution of volume fractions for the local configurations. Our ensemble analysis highlights the importance of collective and global effects in random sphere packings by comparing clusters generated via local rules to random sphere packings and clusters that include collective effects. PMID- 17129153 TI - Tuning the electronic structures of semiconducting SiC nanotubes by N and NHx (x=1,2) groups. AB - We investigate the stable configurations and electronic structures of silicon carbide nanotubes (SiCNTs) decorated by N and NHx (x=1,2) groups by using first principles calculations. We find that these groups can be chemically incorporated into the network of SiCNTs in different ways, accompanied with the formation of N C and N-Si bonds. The adsorbing energy of N and NHx (x=1,2) groups on (5,5) and (8,0) SiCNTs ranges from -1.82 to -7.19 eV. The electronic structures of SiCNTs can be effectively modified by these groups and display diverse characters ranging from semiconducting to semimetallic, depending on the chirality of SiCNTs as well as the way of the incorporation of these functional groups. The relationship between the electronic structures and the configurations of these functionalized SiCNTs is also addressed by performing projected density of states combined with Milliken population analysis. These results are expected to open a way to tune the electronic structures of SiCNTs which may have promising applications in building nanodevices. PMID- 17129154 TI - Determining electronic spectra at interfaces by electronic sum frequency generation: one- and two-photon double resonant oxazine 750 at the air/water interface. AB - The second-order nonlinear electronic spectra were measured for a dye oxazine 750 (OX750) adsorbed at the air/water interface using the multiplex electronic sum frequency generation (ESFG) spectroscopy recently developed by our group. The excitation-wavelength dependence of the ESFG spectrum was investigated, and a global fitting analysis was performed to separate contributions of one- and two photon resonances. The analysis yielded linear interface electronic spectra in the one- and two-photon resonance regions, which can be directly compared to bulk absorption spectra. A two-dimensional plot of the linear interface electronic spectra is newly proposed to graphically represent all the essential information on the electronic structure of interfacial molecules. On this new analytical basis of the ESFG spectroscopy, the spectroscopic properties of OX750 at the interface are discussed. PMID- 17129155 TI - Two bonding configurations of acetylene on Si(001)-(2 x 1): a combined high resolution electron energy loss spectroscopy and density functional theory study. AB - Two coexisting adsorption states of molecularly adsorbed acetylene on the Si(001) (2 x 1) surface have been identified by a combined study based on the high resolution electron energy loss spectroscopy and density functional computations. Seven possible adsorbate-substrate structures are considered theoretically including their full vibrational analysis. Based on a significantly enhanced experimental resolution, the assignment of 15 C2H2- and C2D2-derived vibrational modes identifies a dominant di-sigma bonded molecule adsorbed on top of a single Si-Si dimer. Additionally there is clear evidence for a second minority species which is di-sigma bonded between two Si-Si dimers within the same dimer row (end bridge geometry). The possible symmetries of the adsorbate complexes are discussed based on the specular and off-specular vibrational measurements. They suggest lower than ideal C(2v) and C(s) symmetries for on-top and end-bridge species, respectively. At low coverages the symmetry reductions might be lifted. PMID- 17129156 TI - Investigation of spectral diffusion in ribonuclease by photolabeling of intrinsic aromatic amino acids. AB - Spectral diffusion dynamics in ribonuclease A was observed via the broadening of photochemical holes burned into the absorption spectrum of intrinsic tyrosine residues. Unlike previous results based on hole burning of chromophores in the pockets of heme proteins, where spectral diffusion develops according to a power law in time, the dynamics in ribonuclease follow a logarithmic law. The results suggest that the experiment preferentially labels the tyrosines located on the surface of the protein where the two-level system dynamics of the glass host matrix exert a strong influence. PMID- 17129157 TI - Effective charges along the melting line of colloidal crystals. AB - The shear modulus G of charged colloidal crystals was measured at several constant particle densities n and varying salt concentrations c up to the melting salt concentration cM using torsional resonance spectroscopy. Far from the phase boundary the samples are polycrystalline and the shear modulus stays roughly constant as a function of c. Upon approaching the melting transition an increasing amount of wall based crystal material is formed surrounding a shrinking polycrystalline core and G drops nearly linearly. When the transition is complete G again stays constant. The morphologic transitions may be scaled upon a single master curve. For the polycrystalline morphology, the elastic data are evaluated in terms of a pairwise additive screened Coulomb interaction yielding a particle effective charge Z(G)*. Under de-ionized conditions Z(0,G)* is independent of n and significantly lower than expected from charge renormalization theory. With increasing salt concentration Z(G)* increases. The increase becomes more pronounced at larger n. By extrapolation we further obtain the melting line effective elasticity charge Z(M,G)*. Z(M,G)* shows a steplike increase with increasing nM and cM to values consistent with charge renormalization theory. Interestingly, the increase coincides semi-quantitatively with the one expected from the universal melting line for charged spheres, thus facilitating a consistent description of phase behavior and elasticity over an extended range of the phase diagram. PMID- 17129158 TI - Stochastic simulation of catalytic surface reactions in the fast diffusion limit. AB - The master equation of a lattice gas reaction tracks the probability of visiting all spatial configurations. The large number of unique spatial configurations on a lattice renders master equation simulations infeasible for even small lattices. In this work, a reduced master equation is derived for the probability distribution of the coverages in the infinite diffusion limit. This derivation justifies the widely used assumption that the adlayer is in equilibrium for the current coverages and temperature when all reactants are highly mobile. Given the reduced master equation, two novel and efficient simulation methods of lattice gas reactions in the infinite diffusion limit are derived. The first method involves solving the reduced master equation directly for small lattices, which is intractable in configuration space. The second method involves reducing the master equation further in the large lattice limit to a set of differential equations that tracks only the species coverages. Solution of the reduced master equation and differential equations requires information that can be obtained through short, diffusion-only kinetic Monte Carlo simulation runs at each coverage. These simulations need to be run only once because the data can be stored and used for simulations with any set of kinetic parameters, gas-phase concentrations, and initial conditions. An idealized CO oxidation reaction mechanism with strong lateral interactions is used as an example system for demonstrating the reduced master equation and deterministic simulation techniques. PMID- 17129159 TI - Carbon chains and the (5,5) single-walled nanotube: structure and energetics versus length. AB - Reliable thermochemistry is computed for infinite stretches of pure-carbon materials including acetylenic and cumulenic carbon chains, graphene sheet, and single-walled carbon nanotubes (SWCNTs) by connection to the properties of finite size molecules that grow into the infinitely long systems. Using ab initio G3 theory, the infinite cumulenic chain (:C[double bond]C[double bond]C[double bond]C:) is found to be 1.9+/-0.4 kcal/mol per carbon less stable in free energy at room temperature than the acetylenic chain (.C[triple bond]C-C[triple bond]C.) which is 24.0 kcal/mol less stable than graphite. The difference between carbon carbon triple, double, and single bond lengths (1.257, 1.279, and 1.333 A, respectively) in infinite chains is evident but much less than with small hydrocarbon molecules. These results are used to evaluate the efficacy of similar calculations with the less rigorous PM3 semiempirical method on the (5,5) SWCNT, which is too large to be studied with high-level ab initio methods. The equilibrium electronic energy change for C(g)-->C[infinite (5,5) SWCNT] is -166.7 kcal/mol, while the corresponding free energy change at room temperature is 153.3 kcal/mol (6.7 kcal/mol less stable than graphite). A threefold alternation (6.866, 6.866, and 6.823 A) in the ring diameter of the equilibrium structure of infinitely long (5,5) SWCNT is apparent, although the stability of this structure over the constant diameter structure is small compared to the zero point energy of the nanotube. In general, different (n,m) SWCNTs have different infinite tube energetics, as well as very different energetic trends that vary significantly with length, diameter, and capping. PMID- 17129160 TI - On one-dimensional self-assembly of surfactant-coated nanoparticles. AB - Nanometer-sized metal and semiconductor particles possess novel properties. To fully realize their potential, these nanoparticles need to be fabricated into ordered arrays or predesigned structures. A promising nanoparticle fabrication method is coupled surface passivation and self-assembly of surfactant-coated nanoparticles. Due to the empirical procedure and partially satisfactory results, this method still represents a major challenge to date and its refinement can benefit from fundamental understanding. Existing evidences suggest that the self assembly of surfactant-coated nanoparticles is induced by surfactant-modified interparticle interactions and follows an intrinsic road map such that short one dimensional (1D) chain arrays of nanoparticles occur first as a stable intermediate before further assembly takes place to form higher dimensional close packed superlattices. Here we report a study employing fundamental analyses and Brownian dynamics simulations to elucidate the underlying pair interaction potential that drives the nanoparticle self-assembly via 1D arrays. We find that a pair potential which has a longer-ranged repulsion and reflects the effects of surfactant chain interdigitation on the dynamics is effective in producing and stabilizing nanoparticle chain arrays. The resultant potential energy surface is isotropic for dispersed nanoparticles but becomes anisotropic to favor the growth of linear chain arrays when self-assembly starts. PMID- 17129161 TI - Dynamics of a discotic liquid crystal in the isotropic phase. AB - Optically heterodyne-detected optical Kerr effect (OHD-OKE) experiments are conducted to study the orientational dynamics of a discotic liquid crystal 2,3,6,7,10,11-hexakis(pentyloxy)triphenylene (HPT) in the isotropic phase near the columnar-isotropic (C-I) phase transition. The OHD-OKE signal of HPT is characterized by an intermediate power law t(-0.76+/-0.02) at short times (a few picoseconds), a von Schweidler power law t(-0.26+/-0.01) at intermediate times (hundreds of picoseconds), and an exponential decay at long times (tens of nanoseconds). The exponential decay has Arrhenius temperature dependence. The functional form of the total time dependent decay is identical to the one observed previously for a large number of molecular supercooled liquids. The mode coupling theory schematic model based on the Sjogren [Phys. Rev. A 33, 1254 (1986)] model is able to reproduce the HPT data over a wide range of times from <1 ps to tens of nanoseconds. The studies indicate that the HPT C-I phase transition is a strong first order transition, and the dynamics in the isotropic phase display a complex time dependent profile that is common to other molecular liquids that lack mesoscopic structure. PMID- 17129162 TI - Stoichiometric polyelectrolyte complexes of ionic block copolymers and oppositely charged polyions. AB - Micellization in dilute solutions of diblock copolymers with a polyelectrolyte and a hydrophilic nonionic blocks and oppositely charged polyions is studied using mean-field theory. In aqueous solutions the micelle core consists of the polyelectrolyte complex (PEC) while the corona is formed by hydrophilic blocks of the block copolymers. Describing PEC as a globule in the framework of the Lifshitz [Zh. Eksp. Teor. Fiz. 55, 2408 (1968)] globule theory we calculate the surface tension of the micellar core/solvent interface as a function of the polyion degree of ionization, solvent quality, and concentration of low-molecular mass salt. The equilibrium aggregation number of starlike micelles formed by block copolymers and homopolymers of opposite charge at stoichiometric mixture compositions is found as a function of the system parameters. It is shown that micelles disintegrate upon addition of salt. PMID- 17129163 TI - Morphologies of multicompartment micelles formed by triblock copolymers. AB - Multicompartment micelles are desirable for advanced applications such as drug delivery. Recently, core-shell-corona (CSC) and segmented-worm (SW) micelles formed by ABC triblock terpolymers with three mutually immiscible blocks are observed in experiments. We have performed dissipative particle dynamics simulations to study the effects of molecular architecture, block length, and solution concentration on the morphologies of ABC triblock terpolymers. The formation of CSC and SW micelles for linear and miktoarm star ABC terpolymers is confirmed in this work. In addition, we predict that different multicompartment micellar morphologies (e.g., incomplete skin-layered micelles and segmented worms) can be formed by linear copolymer with different arrangements of the three blocks. PMID- 17129164 TI - Discontinuous molecular dynamics simulation study of polymer collapse. AB - Discontinuous molecular dynamics simulations were used to study the coil-globule transition of a polymer in an explicit solvent. Two different versions of the model were employed, which are differentiated by the nature of monomer-solvent, solvent-solvent, and nonbonded monomer-monomer interactions. For each case, a model parameter lambda determines the degree of hydrophobicity of the monomers by controlling the degree of energy mismatch between the monomers and solvent particles. We consider a lambda-driven coil-globule transition at constant temperature. The simulations are used to calculate average static structure factors, which are then used to determine the scaling exponents of the system in order to determine the theta-point values lambda(theta) separating the coil from the globule state. For each model we construct coil-globule phase diagrams in terms of lambda and the particle density rho. Additionally, we explore for each model the effects of varying the range of the attractive interactions on the phase boundary separating the coil and globule phases. The results are analyzed in terms of a simple Flory-type theory of the collapse transition. PMID- 17129165 TI - Size, shape, and flexibility of RNA structures. AB - Determination of sizes and flexibilities of RNA molecules is important in understanding the nature of packing in folded structures and in elucidating interactions between RNA and DNA or proteins. Using the coordinates of the structures of RNA in the Protein Data Bank we find that the size of the folded RNA structures, measured using the radius of gyration R(G), follows the Flory scaling law, namely, R(G)=5.5N(1/3) A, where N is the number of nucleotides. The shape of RNA molecules is characterized by the asphericity Delta and the shape S parameters that are computed using the eigenvalues of the moment of inertia tensor. From the distribution of Delta, we find that a large fraction of folded RNA structures are aspherical and the distribution of S values shows that RNA molecules are prolate (S>0). The flexibility of folded structures is characterized by the persistence length l(p). By fitting the distance distribution function P(r), that is computed using the coordinates of the folded RNA, to the wormlike chain model we extracted the persistence length l(p). We find that l(p) approximately 1.5N(0.33) A which might reflect the large separation between the free energies that stabilize secondary and tertiary structures. The dependence of l(p) on N implies that the average length of helices should increase as the size of RNA grows. We also analyze packing in the structures of ribosomes (30S, 50S, and 70S) in terms of R(G), Delta, S, and l(p). The 70S and the 50S subunits are more spherical compared to most RNA molecules. The globularity in 50S is due to the presence of an unusually large number (compared to 30S subunit) of small helices that are stitched together by bulges and loops. Comparison of the shapes of the intact 70S ribosome and the constituent particles suggests that folding of the individual molecules might occur prior to assembly. PMID- 17129166 TI - Shear thinning in dilute polymer solutions. AB - We use bead-spring models for a polymer coupled to a solvent described by multiparticle collision dynamics to investigate shear thinning effects in dilute polymer solutions. First, we consider the polymer motion and configuration in a shear flow. For flexible polymer models we find a sharp increase in the polymer radius of gyration and the fluctuations in the radius of gyration at a Weissenberg number approximately 1. We then consider the polymer viscosity and the effect of solvent quality, excluded volume, hydrodynamic coupling between the beads, and finite extensibility of the polymer bonds. We conclude that the excluded volume effect is the major cause of shear thinning in polymer solutions. Comparing the behavior of semiflexible chains, we find that the fluctuations in the radius of gyration are suppressed when compared to the flexible case. The shear thinning is greater and, as the rigidity is increased, the viscosity measurements tend to those for a multibead rod. PMID- 17129167 TI - Analytical theory of hysteresis in ion channels: two-state model. AB - Channel-forming proteins in a lipid bilayer of a biological membrane usually respond to variation of external voltage by changing their conformations. Periodic voltages with frequency comparable with the inverse relaxation time of the protein produce hysteresis in the occupancies of the protein conformations. If the channel conductance changes when the protein jumps between these conformations, hysteresis in occupancies is observed as hysteresis in ion current through the channel. We develop an analytical theory of this phenomenon assuming that the channel conformational dynamics can be described in terms of a two-state model. The theory describes transient behavior of the channel after the periodic voltage is switched on as well as the shape and area of the hysteretic loop as functions of the frequency and amplitude of the applied voltage. The area vanishes as the voltage period T tends to zero and infinity. Asymptotic behaviors of the loop area A in the high- and low-frequency regimes, respectively, are A approximately T and A approximately T(-1). PMID- 17129168 TI - Protein folding using fragment assembly and physical energy function. AB - We perform a systematic study of the effects of sequence-independent backbone interactions and sequence-dependent side-chain interactions on protein folding using fragment assembly and physical energy function. Structures for ten proteins belonging to various structural classes are predicted only with Lennard-Jones interaction between backbone atoms. We find nativelike structures for beta proteins, suggesting that for proteins in this class, the global tertiary structures can be determined mainly by sequence-independent backbone interactions. On the other hand, for alpha proteins, nonlocal hydrophobic side chain interaction is also required to obtain nativelike structures. PMID- 17129169 TI - Pericellular matrilins regulate activation of chondrocytes by cyclic load-induced matrix deformation. AB - Pericellular matrix is at the ideal location to be involved in transmitting mechanical signals from the microenvironment to a cell. We found that changes of the content of matrilins that link various pericellular molecules surrounding chondrocytes affect mechanical stimulation of chondrocyte proliferation and gene expression. Thus, pericellular matrilins may play a role in chondrocyte mechanotransduction. INTRODUCTION: Chondrocytes reside in a capsule of pericellular matrix (chondron), which has been hypothesized to play a critical role in transducing mechanical signals to the cell. In this study, we test the hypothesis that the levels of matrilin (MATN)-1 and -3, major components of the chondrocyte pericellular matrix network, regulate activation of chondrocyte proliferation and differentiation by cyclic load-induced matrix deformation. MATERIALS AND METHODS: Functional matrilins were decreased by expressing a dominant negative mini-MATN in primary chondrocytes or by using MATN1-null chondrocytes. The abundance of matrilins was also increased by expressing a wildtype MATN1 or MATN3 in chondrocytes. Chondrocytes were cultured in a 3D sponge subjected to cyclic deformation at 1 Hz. Chondrocyte gene expression was quantified by real-time RT-PCR and by Western blot analysis. Matrilin pericellular matrix assembly was examined by immunocytochemistry. RESULTS: Elimination of functional matrilins from pericellular matrix abrogated mechanical activation of Indian hedgehog signaling and abolished mechanical stimulation of chondrocyte proliferation and differentiation. Excessive or reduced matrilin content decreased mechanical response of chondrocytes. CONCLUSIONS: Normal content of matrilins is essential to optimal activation of chondrocytes by mechanical signals. Our data suggest that the sensitivity of chondrocytes to the changes in the microenvironment can be adjusted by altering the content of matrilins in pericellular matrix. This finding supports a critical role of pericellular matrix in chondrocyte mechano-transduction and has important implications in cartilage tissue engineering and mechanical adaptation. PMID- 17129170 TI - Hyperostosis-hyperphosphatemia syndrome: a congenital disorder of O-glycosylation associated with augmented processing of fibroblast growth factor 23. AB - Two hyperphosphatemic patients with mutations in GALNT3 showed low intact FGF23 levels with marked increase of processed C-terminal fragments. FGF23 protein has three O-linked glycans and FGF23 with incomplete glycosylation is susceptible to processing. Silencing GALNT3 resulted in enhanced processing of FGF23. Decreased function of FGF23 by enhanced processing is the cause of hyperphosphatemia in patients with GALNT3 mutation. INTRODUCTION: Hyperostosis-hyperphosphatemia syndrome (HHS) is an autosomal recessive entity manifesting as severe hyperphosphatemia associated with episodic bone pain and radiological findings of cortical hyperostosis and periosteal reaction. Persistent hyperphosphatemia is not counterbalanced by PTH or 1,25-dihydroxyvitamin D, posing a mirror image of hypophosphatemic states attributed to increased fibroblast growth factor (FGF)23 activity. MATERIALS AND METHODS: We describe two children with HHS who were found to be homozygous for a mutation in GALNT3 encoding a peptide involved in mucin type O-glycosylation (ppGaNTase-T3). FGF23 levels were evaluated by two ELISAs and Western blotting. FGF23 protein was analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Effect of silencing GALNT3 was evaluated using siRNA in cells transfected with expression vector for FGF23. RESULTS: Both patients had low levels of the full-length FGF23 with markedly augmented amounts of the inactive fragments. Biologically active FGF23 has three O-linked glycans. FGF23 with only one or two O-linked glycans is processed into inactive fragments. Decreasing the expression of the GALNT3 gene by RNA interference resulted in enhanced processing of FGF23. CONCLUSIONS: The primary defect in HHS is impairment of glycosylation of FGF23 resulting from mutations in GALNT3 and leading to augmented processing of FGF23. These changes in FGF23 abolish its phosphaturic effect and lead to severe persistent hyperphosphatemia. This study provides the pathogenetic mechanism of the first mucin-type O-glycosylation defect identified. PMID- 17129171 TI - Paget's disease of bone in the French population: novel SQSTM1 mutations, functional analysis, and genotype-phenotype correlations. AB - Mutation screening of the SQSTM1 gene in 94 French patients with PDB revealed two novel point-mutations (A381V and L413F) and two new compound heterozygous genotypes (P392L/A381V and P392L/A390X). Functional analysis showed an increased level of SQSTM1/p62 protein in PDB patients and truncated forms of the protein encoded by the A390X allele. Clinical data indicate that PDB patients with SQSTM1 mutation are younger at PDB diagnosis and have more extensive bone lesions. INTRODUCTION: Paget's disease of bone (PDB) is a common chronic disease of the skeleton, with a strong genetic component. A recurrent mutation (P392L) was first identified on chromosome 5, in the Sequestosome 1 (SQSTM1) gene. Several other mutations of the SQSTM1 gene have been described in PDB patients, affecting the ubiquitin-associated domain (UBA) of the SQSTM1/p62 protein. The objectives of this study were to evaluate the frequency of the SQSTM1 mutations in French PBD patients, to study the expression of the SQSTM1/p62 protein, and to search for genotype-phenotype correlations. MATERIALS AND METHODS: Blood was obtained from 94 unrelated French PDB patients and 100 controls for mutation screening of exons 7 and 8, encoding for the UBA domain of SQSTM1. Epstein-Barr virus (EBV) immortalized B-cell lymphocytes were established from 13 patients, giving access to functional analysis of the gene and the SQSTM1/p62 expressions using real-time PCR and Western blot. RESULTS: Mutations of the SQSTM1 gene were identified in 12 of the 94 PDB patients (13%). Eight patients carried P392L. Two novel missense mutations were identified: L413F and A381V. This A381V mutation and A390X were found in distinct patients already carriers of P392L. The SQSTM1/p62 protein expression in PDB patients increased when zero, one, or two mutations were present, and SQSTM1 truncated forms were associated with the A390X mutation. The mean age of PDB diagnosis was younger in patients with the SQSTM1 mutation. PDB was more extensive in patients who carried a SQSTM1 mutation. CONCLUSIONS: Mutations of SQSTM1 are present in the French population. PDB patients with and without the SQSTM1 mutation have an increased level of SQSTM1/p62, caused by overproduction of the protein, probably involved in the pathophysiology of PDB. The presence of the SQSTM1 mutation may be a worsening factor for PDB. PMID- 17129172 TI - Exercise when young provides lifelong benefits to bone structure and strength. AB - Short-term exercise in growing rodents provided lifelong benefits to bone structure, strength, and fatigue resistance. Consequently, exercise when young may reduce the risk for fractures later in life, and the old exercise adage of "use it or lose it" may not be entirely applicable to the skeleton. INTRODUCTION: The growing skeleton is most responsive to exercise, but low-trauma fractures predominantly occur in adults. This disparity has raised the question of whether exercised-induced skeletal changes during growth persist into adulthood where they may have antifracture benefits. This study investigated whether brief exercise during growth results in lifelong changes in bone quantity, structure, quality, and mechanical properties. MATERIALS AND METHODS: Right forearms of 5 week-old Sprague-Dawley rats were exercised 3 days/week for 7 weeks using the forearm axial compression loading model. Left forearms were internal controls and not exercised. Bone quantity (mineral content and areal density) and structure (cortical area and minimum second moment of area [I(MIN)]) were assessed before and after exercise and during detraining (restriction to home cage activity). Ulnas were removed after 92 weeks of detraining (at 2 years of age) and assessed for bone quality (mineralization) and mechanical properties (ultimate force and fatigue life). RESULTS: Exercise induced consistent bone quantity and structural adaptation. The largest effect was on I(MIN), which was 25.4% (95% CI, 15.6 35.3%) greater in exercised ulnas compared with nonexercised ulnas. Bone quantity differences did not persist with detraining, whereas all of the absolute difference in bone structure between exercised and nonexercised ulnas was maintained. After detraining, exercised ulnas had 23.7% (95% CI, 13.0-34.3%) greater ultimate force, indicating enhanced bone strength. However, exercised ulnas also had lower postyield displacement (-26.4%; 95% CI, -43.6% to -9.1%), indicating increased brittleness. This resulted from greater mineralization (0.56%; 95% CI, 0.12-1.00%), but did not influence fatigue life, which was 10 fold greater in exercised ulnas. CONCLUSIONS: These data indicate that exercise when young can have lifelong benefits on bone structure and strength, and potentially, fracture risk. They suggest that the old exercise adage of "use it or lose it" may not be entirely applicable to the skeleton and that individuals undergoing skeletal growth should be encouraged to perform impact exercise. PMID- 17129173 TI - Intermolecular interactions in biomolecular systems examined by mass spectrometry. AB - With the development of electrospray and matrix-assisted laser desorption ionization, mass spectrometry (MS) evolved into a powerful tool in the field of biochemistry. Whereas MS is primarily analytical in nature, an increasing number of MS research groups employ the method to address fundamental biochemical questions. Probing the interaction of noncovalently bound molecules in the mass spectrometer is one of the most interesting MS-based experiments possible today, with the potential of making a significant contribution to the basic understanding of the structure and function of biochemical complexes. Here we review a number of current research efforts employing primarily MS techniques to investigate intermolecular interactions in biochemical systems. Examples chosen include the interaction of biomolecules with solvent molecules; interactions between nucleic-acid molecules, in particular, interactions in duplex and quadruplex structures; and interactions between proteins involved in neurodegenerative diseases. Finally we conclude by presenting a few examples of very large biomolecular assemblies in the mega-Dalton range analyzed by MS. PMID- 17129174 TI - Assessing public health emergency preparedness: concepts, tools, and challenges. AB - Policymakers are increasingly seeking to determine whether the federal government's recent investments in public health preparedness have left the public health system better prepared to respond to large-scale public health emergencies. Yet, there remain questions about how to define "public health emergency preparedness," how much preparedness is enough, and how preparedness can be measured and assessed. This chapter identifies the key challenges associated with measuring public health preparedness and reviews approaches currently in use. We also identify some emerging measurement techniques that might help address some of these challenges. PMID- 17129175 TI - Immunobiology of allogeneic hematopoietic stem cell transplantation. AB - Allogeneic hematopoietic stem cell transplantation (HSCT) has evolved into an effective adoptive cellular immunotherapy for the treatment of a number of cancers. The immunobiology of allogeneic HSCT is unique in transplantation in that it involves potential immune recognition and attack between both donor and host. Much of the immunobiology of allogeneic HSCT has been gleaned from preclinical models and correlation with clinical observations. We review our current understanding of some of the issues that affect the success of this therapy, including host-versus-graft (HVG) reactions, graft-versus-host disease (GVHD), graft-versus-tumor (GVT) activity, and restoration of functional immunity to prevent transplant-related opportunistic infections. We also review new strategies to optimize the GVT and improve overall immune function while reducing GVHD and graft rejection. PMID- 17129176 TI - Differential gene expression in cultured osteoblasts and bone marrow stromal cells from patients with Paget's disease of bone. AB - Paget's disease is a focal condition of bone. To study changes in cells within pagetic lesions, we cultured osteoblasts and stromal cells from 22 patients and compared gene expression in these cells to cells from healthy bone. We identified several differentially regulated genes, and we suggest that these changes could lead to the formation of the lesions. INTRODUCTION: Paget's disease is a focal condition of bone of unknown cause. Although it is regarded as primarily an osteoclast disorder, the tight coupling of the activity of osteoclasts and osteoblasts suggests that the osteoblast could play a key role in its pathogenesis. The aim of the study was to identify possible changes in pagetic osteoblasts and stromal cells that might contribute to the development of pagetic lesions. MATERIALS AND METHODS: Candidate genes were identified based on known bone cell regulators, supplemented with microarray analysis. Gene expression was determined by real-time PCR in primary cultures of osteoblasts and bone marrow stromal cells from pagetic patients and control subjects. Concentrations of secreted proteins were determined by ELISA. RESULTS: Dickkopf1 mRNA and protein levels were increased in both pagetic osteoblast and stromal cell cultures, and interleukin (IL)-1 and IL-6 were overexpressed in pagetic osteoblasts. These changes parallel recent findings in myeloma bone disease, which shares some clinical similarities with Paget's disease. Alkaline phosphatase was overexpressed, and bone sialoprotein and osteocalcin were underexpressed in pagetic osteoblasts, consistent with their circulating levels in pagetic patients. It is hypothesized that overexpression of Dickkopf1, IL-1, and IL-6 would result in stimulation of osteoclast proliferation and inhibition of osteoblast growth, leading to the development of the characteristic lytic bone lesions. By stimulating osteoblast differentiation, Dickkopf1 and IL-6 may also promote mineralization, leading to the conversion of lytic lesions to sclerotic. CONCLUSIONS: These findings suggest that dysregulated gene expression in pagetic osteoblasts could cause the changes in bone cell number and function characteristic of Paget's disease. PMID- 17129177 TI - Changes to osteoporosis prevalence according to method of risk assessment. AB - The impact of clinical risk factor-based absolute risk methods on the prevalence of high risk for osteoporotic fracture is unknown. We applied absolute risk methods to 6646 subjects and found that the prevalence of elderly women deemed to be at high risk increased substantially, whereas the overall prevalence was highly dependent on the threshold used to designate high risk. INTRODUCTION: Many groups have advocated using absolute risk methods that incorporate clinical risk factors to target patients for osteoporosis therapy. We examined how the application of such absolute risk classification systems influences the prevalence of those considered to be at high risk for osteoporotic fracture and compared these systems to one based solely on BMD. MATERIALS AND METHODS: Using 6646 subjects from the Canadian Multicentre Osteoporosis Study (CaMos), a prospective, randomly selected, population-based cohort, we assessed three different systems for determining prevalence of high risk for osteoporotic fracture: a BMD-based system; a simplified risk factor system incorporating age, sex, BMD, and two clinical risk factors; and a comprehensive system, incorporating age, sex, BMD, and seven clinical risk factors. The 10-year absolute risks of incident fragility fracture were compared across systems using three different high-risk thresholds. RESULTS: The prevalence of a T score < or = -2.5 was 18.8% (95% CI: 17.7-19.9%) in women and 3.9% (95% CI: 3.0-4.7%) in men. Using a 15% 10-year risk of fracture threshold, the prevalence of women at high risk increased to 46.9% (95% CI: 45.4-48.4) and 42.5% (95% CI: 41.1-43.9) when the comprehensive and simplified risk factor classification systems were used, respectively. Using a 25% 10-year absolute risk threshold, the prevalence of high risk was similar to that of the BMD-based system, whereas the 20% threshold gave intermediate rates. All thresholds analyzed resulted in an increased prevalence of older women at high risk for fracture, whereas only the 15% 10-year risk of fracture threshold resulted in an increase in the prevalence of men at high risk. CONCLUSIONS: The application of risk factor-based systems results in an increased prevalence of older women at high risk. The prevalence of individuals at high risk may increase with changes to the methods used to determine those who are eligible for therapy. These data have important implications for the pattern of care and costs of treating osteoporotic fractures. PMID- 17129178 TI - Marked disturbance of calcium homeostasis in mice with targeted disruption of the Trpv6 calcium channel gene. AB - We report the phenotype of mice with targeted disruption of the Trpv6 (Trpv6 KO) epithelial calcium channel. The mice exhibit disordered Ca(2+) homeostasis, including defective intestinal Ca(2+) absorption, increased urinary Ca(2+) excretion, decreased BMD, deficient weight gain, and reduced fertility. Although our Trpv6 KO affects the closely adjacent EphB6 gene, the phenotype reported here is not related to EphB6 dysfunction. INTRODUCTION: The mechanisms underlying intestinal Ca(2+) absorption are crucial for overall Ca(2+) homeostasis, because diet is the only source of all new Ca(2+) in the body. Trpv6 encodes a Ca(2+) permeable cation channel responsible for vitamin D-dependent intestinal Ca(2+) absorption. Trpv6 is expressed in the intestine and also in the skin, placenta, kidney, and exocrine organs. MATERIALS AND METHODS: To determine the in vivo function of TRPV6, we generated mice with targeted disruption of the Trpv6 (Trpv6 KO) gene. RESULTS: Trpv6 KO mice are viable but exhibit disordered Ca(2+) homeostasis, including a 60% decrease in intestinal Ca(2+) absorption, deficient weight gain, decreased BMD, and reduced fertility. When kept on a regular (1% Ca(2+)) diet, Trpv6 KO mice have deficient intestinal Ca(2+) absorption, despite elevated levels of serum PTH (3.8-fold) and 1,25-dihydroxyvitamin D (2.4-fold). They also have decreased urinary osmolality and increased Ca(2+) excretion. Their serum Ca(2+) is normal, but when challenged with a low (0.25%) Ca(2+) diet, Trpv6 KO mice fail to further increase serum PTH and vitamin D, ultimately developing hypocalcemia. Trpv6 KO mice have normal urinary deoxypyridinoline excretion, although exhibiting a 9.3% reduction in femoral mineral density at 2 months of age, which is not restored by treatment for 1 month with a high (2%) Ca(2+) "rescue" diet. In addition to their deranged Ca(2+) homeostasis, the skin of Trpv6 KO mice has fewer and thinner layers of stratum corneum, decreased total Ca(2+) content, and loss of the normal Ca(2+) gradient. Twenty percent of all Trpv6 KO animals develop alopecia and dermatitis. CONCLUSIONS: Trpv6 KO mice exhibit an array of abnormalities in multiple tissues/organs. At least some of these are caused by tissue-specific mechanisms. In addition, the kidneys and bones of Trpv6 KO mice do not respond to their elevated levels of PTH and 1,25 dihydroxyvitamin D. These data indicate that the TRPV6 channel plays an important role in Ca(2+) homeostasis and in other tissues not directly involved in this process. PMID- 17129179 TI - Osteosarcoma and teriparatide? PMID- 17129180 TI - TSLP: an epithelial cell cytokine that regulates T cell differentiation by conditioning dendritic cell maturation. AB - Dendritic cells (DCs) are professional antigen-presenting cells that have the ability to sense infection and tissue stress, sample and present antigen to T lymphocytes, and induce different forms of immunity and tolerance. The functional versatility of DCs depends on their remarkable ability to translate collectively the information from both the invading microbes and their resident tissue microenvironments and then make an appropriate immune response. Recent progress in understanding TLR biology has illuminated the mechanisms by which DCs link innate and adaptive antimicrobial immune responses. However, how tissue microenvironments shape the function of DCs has remained elusive. Recent studies of TSLP (thymic stromal lymphopoietin), an epithelial cell-derived cytokine that strongly activates DCs, provide evidence at a molecular level that epithelial cells/tissue microenvironments directly communicate with DCs. We review recent progress on how TSLP expressed within thymus and peripheral lymphoid and nonlymphoid tissues regulates DC-mediated central tolerance, peripheral T cell homeostasis, and inflammatory Th2 responses. PMID- 17129181 TI - Improving T cell therapy for cancer. AB - Adoptive transfer of antigen-specific T lymphocytes is a powerful therapy for the treatment of opportunistic disease and some virus-associated malignancies such as Epstein-Barr virus-positive post-transplant lymphoproliferative disease. However, this strategy has been less successful in patients with nonviral cancers owing to their many and varied immune evasion mechanisms. These mechanisms include downregulation of target antigens and antigen-presenting machinery, secretion of inhibitory cytokines, and recruitment of regulatory immune cells to the tumor site. With increased understanding of the tumor microenvironment and the behavior and persistence of ex vivo-manipulated, adoptively transferred T cells, two novel approaches for increasing the efficacy of T cell therapy have been proposed. The first involves genetic modification of tumor-specific T cells to improve their biological function, for example by augmenting their ability to recognize tumor cells or their resistance to tumor-mediated immunosuppression. The second requires modifications to the host environment to improve the homeostatic expansion of infused T cells or to eliminate inhibitory T cell subsets. In this review, we discuss current, promising strategies to improve adoptive T cell therapy for the treatment of cancer. PMID- 17129182 TI - Effector and memory CTL differentiation. AB - Technological advances in recent years have allowed for an ever-expanding ability to analyze and quantify in vivo immune responses. MHC tetramers, intracellular cytokine staining, an increasing repertoire of transgenic and "knockout" mice, and the detailed characterization of a variety of infectious models have all facilitated more precise and definitive analyses of the generation and function of cytotoxic T lymphocytes (CTL). Understanding the mechanisms behind the differentiation of effector and memory CTL is of increasing importance to develop vaccination strategies against a variety of established and emerging infectious diseases. This review focuses on recent advances in our understanding of how effector and memory CTL differentiate and survive in vivo in response to viral or bacterial infection. PMID- 17129183 TI - Intracellular targets of matrix metalloproteinase-2 in cardiac disease: rationale and therapeutic approaches. AB - A new paradigm of matrix metalloproteinase-2 (MMP-2) action in the heart undergoing oxidative stress has emerged. Although best known for its role in the proteolysis of extracellular protein targets, MMP-2 is also localized to the sarcomere within the cardiomyocyte. Oxidative stress activates full-length MMP-2 without need for proteolytic processing and inactivates an endogenous inhibitor, tissue inhibitor of metalloproteinase-4. MMP-2 proteolyzes specific targets within the cell to cause acute, reversible contractile dysfunction. Inhibitors of MMPs are discussed and their possible use for the therapy of acute heart injury caused by oxidative stress is examined. PMID- 17129184 TI - Separate neural processing of timbre dimensions in auditory sensory memory. AB - Timbre is a multidimensional perceptual attribute of complex tones that characterizes the identity of a sound source. Our study explores the representation in auditory sensory memory of three timbre dimensions (acoustically related to attack time, spectral centroid, and spectrum fine structure), using the mismatch negativity (MMN) component of the auditory event related potential. MMN is elicited by a discriminable change in a sound sequence and reflects the detection of the discrepancy between the current stimulus and traces in auditory sensory memory. The stimuli used in the present study were carefully controlled synthetic tones. MMNs were recorded after changes along each of the three timbre dimensions and their combinations. Additivity of unidimensional MMNs and dipole modeling results suggest partially separate MMN generators for different timbre dimensions, reflecting their mainly separate processing in auditory sensory memory. The results expand to timbre dimensions a property of separation of the representation in sensory memory that has already been reported between basic perceptual attributes (pitch, loudness, duration, and location) of sound sources. PMID- 17129185 TI - The human striatum is necessary for responding to changes in stimulus relevance. AB - Various lines of evidence suggest that the striatum is implicated in cognitive flexibility. The neuropsychological evidence has, for the most part, been based on research with patients with Parkinson's disease, which is accompanied by chemical disruption of both the striatum and the prefrontal cortex. The present study examined this issue by testing patients with focal lesions of the striatum on a task measuring two forms of cognitive switching. Patients with striatal, but not frontal lobe lesions, were impaired in switching between concrete sensory stimuli. By contrast, both patient groups were unimpaired when switching between abstract task rules relative to baseline nonswitch trials. These results reveal a dissociation between two distinct forms of cognitive flexibility, providing converging evidence for a role of the striatum in flexible control functions associated with the selection of behaviorally relevant stimuli. PMID- 17129186 TI - Anatomical substrates of visual and auditory miniature second-language learning. AB - Longitudinal changes in brain activity during second language (L2) acquisition of a miniature finite-state grammar, named Wernickese, were identified with functional magnetic resonance imaging (fMRI). Participants learned either a visual sign language form or an auditory-verbal form to equivalent proficiency levels. Brain activity during sentence comprehension while hearing/viewing stimuli was assessed at low, medium, and high levels of proficiency in three separate fMRI sessions. Activation in the left inferior frontal gyrus (Broca's area) correlated positively with improving L2 proficiency, whereas activity in the right-hemisphere (RH) homologue was negatively correlated for both auditory and visual forms of the language. Activity in sequence learning areas including the premotor cortex and putamen also correlated with L2 proficiency. Modality specific differences in the blood oxygenation level-dependent signal accompanying L2 acquisition were localized to the planum temporale (PT). Participants learning the auditory form exhibited decreasing reliance on bilateral PT sites across sessions. In the visual form, bilateral PT sites increased in activity between Session 1 and Session 2, then decreased in left PT activity from Session 2 to Session 3. Comparison of L2 laterality (as compared to L1 laterality) in auditory and visual groups failed to demonstrate greater RH lateralization for the visual versus auditory L2. These data establish a common role for Broca's area in language acquisition irrespective of the perceptual form of the language and suggest that L2s are processed similar to first languages even when learned after the "critical period." The right frontal cortex was not preferentially recruited by visual language after accounting for phonetic/structural complexity and performance. PMID- 17129187 TI - "What" and "where" in word reading: ventral coding of written words revealed by parietal atrophy. AB - The visual system of literate adults develops a remarkable perceptual expertise for printed words. To delineate the aspects of this competence intrinsic to the occipitotemporal "what" pathway, we studied a patient with bilateral lesions of the occipitoparietal "where" pathway. Depending on critical geometric features of the display (rotation angle, letter spacing, mirror reversal, etc.), she switched from a good performance, when her intact ventral pathway was sufficient to encode words, to severely impaired reading, when her parietal lesions prevented the use of alternative reading strategies as a result of spatial and attentional impairments. In particular, reading was disrupted (a) by rotating word by more than 50 degrees , providing an approximation of the invariance range for words encoding in the ventral pathway; (b) by separating letters with double spaces, revealing the limits of letter grouping into perceptual wholes; (c) by mirror reversing words, showing that words escape the default mirror-invariant representation of visual objects in the ventral pathway. Moreover, because of her parietal lesions, she was unable to discriminate mirror images of common objects, although she was excellent with reversible pseudowords, confirming that the breaking of mirror symmetry was intrinsic to the occipitotemporal cortex. Thus, charting the display conditions associated with preserved or impaired performance allowed us to infer properties of word coding in the normal ventral pathway and to delineate the roles of the parietal lobes in single-word recognition. PMID- 17129188 TI - Repetition suppression for spoken sentences and the effect of task demands. AB - We examined whether the repeated processing of spoken sentences is accompanied by reduced bold oxygenation level-dependent response (repetition suppression) in regions implicated in sentence comprehension and whether the magnitude of such suppression depends on the task under which the sentences are comprehended or on the complexity of the sentences. We found that sentence repetition was associated with repetition suppression in temporal regions, independent of whether participants judged the sensibility of the statements or listened to the statements passively. In contrast, repetition suppression in inferior frontal regions was found only in the context of the task demanding active judgment. These results suggest that repetition suppression in temporal regions reflects facilitation of sentence comprehension processing per se, whereas in frontal regions it reflects, at least in part, easier execution of specific psycholinguistic judgments. PMID- 17129189 TI - The impact of proficiency on syntactic second-language processing of German and Italian: evidence from event-related potentials. AB - The present study investigated the role of proficiency in late second-language (L2) processing using comparable stimuli in German and Italian. Both sets of stimuli consisted of simple active sentences including a word category violation, a morphosyntactic agreement violation, or a combination of the two. Four experiments were conducted to study high- and low-proficiency L2 learners of German as well as high- and low-proficiency L2 learners of Italian. High proficiency L2 learners in both languages showed the same event-related potential (ERP) components as native speakers for all syntactic violations. For the word category violation, they displayed an early anterior negativity (ELAN), an additional negativity reflecting reference-related processes, and a late P600 evidencing processes of reanalysis. For the processing of the morphosyntactic error, an anterior negativity (LAN) and a P600 were observed, whereas for the combined violation, the same ERP components were found as in the pure category violation. In high-proficiency L2 learners, the timing of the processing steps was equivalent to that of native speakers, although some amplitude differences were present. Low-proficiency L2 learners, however, showed qualitative differences in the agreement violation characterized by the absence of the LAN and quantitative differences reflected in a delayed P600 in every violation condition. These findings emphasize that with a high proficiency, late L2 learners can indeed show native-like neural responses with the timing approximating that of native speakers. This challenges the idea that there are fundamental differences in language processing in the brain between natives and late L2 learners. PMID- 17129190 TI - Imaging informational conflict: a functional magnetic resonance imaging study of numerical stroop. AB - We employed a parametric version of the comparison Stroop paradigm to investigate the processing of numerical magnitude and physical size under task-relevant and irrelevant conditions to investigate two theoretical issues: (1) What is the neural fate of task-irrelevant information? (2) What is the neural basis of the resolution of the conflict between task-relevant and -irrelevant information? We show in 18 healthy adults that numerical magnitudes of numbers call for higher processing requirements than physical sizes. The enhanced activation elicited by numerical magnitudes is not modulated by task relevance, indicating autonomous processing. Moreover, the normal behavioral distance effect when the numerical dimension is task relevant and reversed distance effect when it is not show that autonomous processing fully encodes numerical magnitudes. Conflict trials elicited greater activation in bilateral inferior frontal gyri, right middle frontal gyri, and right superior frontal gyri. We postulate two sources to the conflict, namely, at cognitive and response levels. PMID- 17129191 TI - Deficits in movement planning and intrinsic coordinate control in ideomotor apraxia. AB - Two central issues in the field of motor control are the coordinate frame in which movements are controlled and the distinction between movement planning and online correction. In this study we used these issues to frame several hypotheses about the deficits underlying ideomotor apraxia (IMA). In particular, we examined whether ideomotor apraxics exhibited (1) deficits in movement control in intrinsic (body relative) coordinates with better control in extrinsic (workspace relative) coordinates, (2) deficits in movement planning that are compensated for by an overreliance on online correction, or (3) both deficits. Patients with IMA and two comparison groups performed movement tasks that relied preferentially on either intrinsic or extrinsic coordinate control when online correction was either possible or impossible. Participants performed posture imitation and grasp imitation movements to body- and object-relative end positions in the presence or absence of visual feedback. Consistent with the intrinsic coordinate control hypothesis, patients with IMA showed a significantly greater disparity than the other two groups between movements made to body-relative and object-relative targets as well as between imitation of meaningless postures and grasping. Consistent with the correction overreliance hypothesis, the IMA group was more disrupted than the other groups by the removal of vision. Thus, IMA patients exhibit behavioral patterns consistent with both deficient intrinsic coordinate control and overreliance upon visual feedback. Finally, lesion analysis suggests that damage to the left inferior parietal lobe (Brodmann's areas 39 and 40) may play a key role in both behavioral deficits. PMID- 17129192 TI - What neuroscience can tell about intuitive processes in the context of perceptual discovery. AB - According to the Oxford English Dictionary, intuition is "the ability to understand or know something immediately, without conscious reasoning." Most people would agree that intuitive responses appear as ideas or feelings that subsequently guide our thoughts and behaviors. It is proposed that people continuously, without conscious attention, recognize patterns in the stream of sensations that impinge upon them. What exactly is being recognized is not clear yet, but we assume that people detect potential content based on only a few aspects of the input (i.e., the gist). The result is a vague perception of coherence which is not explicitly describable but instead embodied in a "gut feeling" or an initial guess, which subsequently biases thought and inquiry. To approach the nature of intuitive processes, we used functional magnetic resonance imaging when participants were working at a modified version of the Waterloo Gestalt Closure Task. Starting from our conceptualization that intuition involves an informed judgment in the context of discovery, we expected activation within the median orbito-frontal cortex (OFC), as this area receives input from all sensory modalities and has been shown to be crucially involved in emotionally driven decisions. Results from a direct contrast between intuitive and nonintuitive judgments, as well as from a parametric analysis, revealed the median OFC, the lateral portion of the amygdala, anterior insula, and ventral occipito-temporal regions to be activated. Based on these findings, we suggest our definition of intuition to be promising and a good starting point for future research on intuitive processes. PMID- 17129193 TI - A neurolinguistic model of grammatical construction processing. AB - One of the functions of everyday human language is to communicate meaning. Thus, when one hears or reads the sentence, "John gave a book to Mary," some aspect of an event concerning the transfer of possession of a book from John to Mary is (hopefully) transmitted. One theoretical approach to language referred to as construction grammar emphasizes this link between sentence structure and meaning in the form of grammatical constructions. The objective of the current research is to (1) outline a functional description of grammatical construction processing based on principles of psycholinguistics, (2) develop a model of how these functions can be implemented in human neurophysiology, and then (3) demonstrate the feasibility of the resulting model in processing languages of typologically diverse natures, that is, English, French, and Japanese. In this context, particular interest will be directed toward the processing of novel compositional structure of relative phrases. The simulation results are discussed in the context of recent neurophysiological studies of language processing. PMID- 17129194 TI - Time course of brain activity during change blindness and change awareness: performance is predicted by neural events before change onset. AB - People often remain "blind" to visual changes occurring during a brief interruption of the display. The processing stages responsible for such failure remain unresolved. We used event-related potentials to determine the time course of brain activity during conscious change detection versus change blindness. Participants saw two successive visual displays, each with two faces, and reported whether one of the faces changed between the first and second displays. Relative to blindness, change detection was associated with a distinct pattern of neural activity at several successive processing stages, including an enhanced occipital P1 response and a sustained frontal activity (CNV-like potential) after the first display, before the change itself. The amplitude of the N170 and P3 responses after the second visual display were also modulated by awareness of the face change. Furthermore, a unique topography of event-related potential activity was observed during correct change and correct no-change reports, but not during blindness, with a recurrent time course in the stimulus sequence and simultaneous sources in the parietal and temporo-occipital cortex. These results indicate that awareness of visual changes may depend on the attentional state subserved by coordinated neural activity in a distributed network, before the onset of the change itself. PMID- 17129195 TI - When gaze turns into grasp. AB - Previous research has provided evidence for a neural system underlying the observation of another person's hand actions. Is the neural system involved in this capacity also important in inferring another person's motor intentions toward an object from their eye gaze? In real-life situations, humans use eye movements to catch and direct the attention of others, often without any accompanying hand movements or speech. In an event-related functional magnetic resonance imaging study, subjects observed videos showing a human model either grasping a target object (grasping condition) or simply gazing (gaze condition) at the same object. These two conditions were contrasted with each other and against a control condition in which the human model was standing behind the object without performing any gazing or grasping action. The results revealed activations within the dorsal premotor cortex, the inferior frontal gyrus, the inferior parietal lobule, and the superior temporal sulcus in both "grasping" and "gaze" conditions. These findings suggest that signaling the presence of an object through gaze elicits in an observer a similar neural response to that elicited by the observation of a reach-to-grasp action performed on the same object. PMID- 17129196 TI - Patterns of regional brain hypometabolism associated with knowledge of semantic features and categories in Alzheimer's disease. AB - The study of semantic memory in patients with Alzheimer's disease (AD) has raised important questions about the representation of conceptual knowledge in the human brain. It is still unknown whether semantic memory impairments are caused by localized damage to specialized regions or by diffuse damage to distributed representations within nonspecialized brain areas. To our knowledge, there have been no direct correlations of neuroimaging of in vivo brain function in AD with performance on tasks differentially addressing visual and functional knowledge of living and nonliving concepts. We used a semantic verification task and resting 18-fluorodeoxyglucose positron emission tomography in a group of mild to moderate AD patients to investigate this issue. The four task conditions required semantic knowledge of (1) visual, (2) functional properties of living objects, and (3) visual or (4) functional properties of nonliving objects. Visual property verification of living objects was significantly correlated with left posterior fusiform gyrus metabolism (Brodmann's area [BA] 37/19). Effects of visual and functional property verification for non-living objects largely overlapped in the left anterior temporal (BA 38/20) and bilateral premotor areas (BA 6), with the visual condition extending more into left lateral precentral areas. There were no associations with functional property verification for living concepts. Our results provide strong support for anatomically separable representations of living and nonliving concepts, as well as visual feature knowledge of living objects, and against distributed accounts of semantic memory that view visual and functional features of living and nonliving objects as distributed across a common set of brain areas. PMID- 17129197 TI - Precursors of insight in event-related brain potentials. AB - Event-related potentials (ERPs) were investigated to find precursors of insightful behavior. Participants had to process successive pairs in strings of digits to obtain a final response in each trial. Within the sequence of five responses required in each trial, the last two responses mirrored the two preceding ones. This hidden regularity, allowing for shortcutting each trial from five to two responses, was discovered by 6 out of 26 participants. Both groups, solvers and nonsolvers, implicitly learned the regularity, reflected by faster responses to the repeated, predictable responses, but this differential effect was larger in solvers, whereas nonsolvers became unspecifically faster with all responses. Several ERP components were larger in solvers than in nonsolvers from the outset: slow positive wave, frontocentral P3a, anterior N1 to those digits that triggered the critical repeating responses, and P3b to the digit that evoked the immediately repeating response. Being already present in the first block, these effects were early precursors of insightful behavior. This early occurrence suggests that participants who will gain insight may be distinguished beforehand by their individual characteristics. PMID- 17129198 TI - Proactive interference as a result of persisting neural representations of previously learned motor skills in primary motor cortex. AB - Learning to control movements in different dynamic environments is marked by proactive interference; learning a first skill interferes with the subsequent learning of a second one. The neural basis of this effect is poorly understood. We tested the idea that proactive interference results from persisting neural representations of previously learned skills in the primary motor cortex (M1). We used repetitive transcranial magnetic stimulation (rTMS) of M1 to disrupt retention of a recently learned motor skill. If interference results from the retention of this skill then its disruption should be associated with reduced interference. Subjects reached to targets while interacting with a robotic arm that applied force fields to the limb. Fifteen minutes of 1-Hz rTMS to M1 impaired the retention of a first force field, and more importantly, reduced proactive interference when subjects learned a second one. Our findings suggest that retention and interference are linked at the level of M1. PMID- 17129199 TI - Disability and anticipatory discourse: the interconnectedness of local and global aspects of talk. AB - In this paper, I use nexus analysis framework to examine how one quadriplegic man from Oman, named Yahya, directs the course of his present (and his future) through anticipatory discourse. In particular, I analyze conversations between Yahya and me regarding his future as they relate to actions I undertake on his behalf to effect a social change: to secure Yahya, an unmarried man, a permit to hire his own resident nurse/assistant (permits for such hirings are reserved only for married couples in Oman). I demonstrate how Yahya influences me to follow his agenda through constructing a helpless identity in narrative discourse. Reciprocally, I suggest that through the actions Yahya's caregivers subsequently undertake, they succeed in giving him a sense of control over an important aspect of his life (being able to hire his own assistant), and I demonstrate how Yahya subsequently displays agentivity in his narratives. I thus illustrate the interconnectedness of Yahya's anticipatory discourse and his caregivers' (macro level) actions that cause societal change, as well as the interconnectedness of these actions and the kinds of selves Yahya constructs through anticipatory discourse in private (micro-level) interactions. PMID- 17129200 TI - Patient narratives: a micro-interactional analysis. AB - The use of narrative as a therapeutic tool is well established in many forms of counseling and psychotherapy. Similarly, it forms a significant element of many holistic consultation approaches. The value of patient narratives are recognized by many practitioners working in primary care too, but opportunities for its utilization can be limited by the practicalities of working within conventional healthcare structures. This article presents a micro-interactional analysis of the ways in which patients routinely frame and self-manage their narratives in the context of primary care encounters, and explores some of the behavioral motifs utilized by practitioners in the management of narrative-based interactions. PMID- 17129201 TI - Unethical bunglers or humane professionals? Discussions in the media of end-of life treatment decisions. AB - We studied media discussions of end-of-life decision making to find out how the doctor-patient relationship is portrayed and what types of positions the media takes and how it is represented by other discussion participants in this context. The significance of the end-of-life decision making is related to the fact that it forms a contested site for the realization of patient autonomy. Our material consisted of two newspaper discussions and one television program. In all three samples, the consumerist physician-patient relationship model was promoted in the context of end-of-life decision making by the media representatives and the laypeople (patients, relatives, and a patient advocate). In the first, the media representatives make use of the romance narrative, which presents physicians as villains, and constructs patients as knowledgeable and competent 'ordinary heroes'. This justifies public scrutiny of healthcare. In the second sample, the relatives stress their competence but now by appealing to their knowledge and caring for the patient. This promotes their right to make decisions for patients. In the third sample, the journalist again pictures patients as 'ordinary heroes' who acquire their knowledge from the media. Patients are now empowered as consumers of healthcare. In conclusion, the consumerist doctor-patient relationship is promoted and the media's position as the source of information and as patient advocate is stabilized. PMID- 17129202 TI - The rhetorical construction of ethical positions: policy recommendations for nontherapeutic genetic testing in childhood. AB - Nontherapeutic genetic testing in childhood raises many ethical concerns within and beyond the clinic. We examine six key position statements from Canada, the United Kingdom, and the United States that present ethical guidelines for good practice in clinical nontherapeutic childhood testing. Using a discourse-analytic perspective that focuses on the use of rhetorical contrasts, we identify how these statements argue for recommendations with distinctly different modalities for different types of nontherapeutic genetic testing. This comes about because of the interaction between a number of contrastive descriptions. It is dependent on how the genetic information resulting from testing is differentiated on a cline of seriousness, how such an evaluation is premised on a network of assumptions about the status of reproduction in people's lives, and the related selective deployment of ethical principles that foregrounds the self over others. PMID- 17129203 TI - Constructing 'expertness': a novice pharmacist's development of interactional competence in patient consultations. AB - It has been argued that being able to utilize expertise in discourse is as important as having expertise (C. Candlin and S. Candlin 2002). From a constructivist perspective, knowledgeability does not exist outside of social interaction: being expert involves performing knowledge and skills effectively in social practices (cf. Butler 1990; Lave and Wenger 1991). It is, then, crucial that novice professionals develop the interactional competence to construct themselves as experts in interaction. This study brings together perspectives from research on expert-lay communication, social interaction, and situated learning to examine how a pharmacy intern performed his professional knowledge in the practice of patient consultation over time. Using conversation analysis complemented by the interactional competence framework (Hall 1995; Young 2002), I demonstrate that as the novice pharmacist gained more experience over the course of one internship, he was able to employ professional knowledge in ways that were sensitive to the sequential organization of talk, and to share the patients' lifeworld perspectives (Mishler 1984) more effectively, thus successfully managing the transactional and interpersonal functions of the consultations. This study contributes to research on expert-lay interaction, particularly in pharmacy, and to the under-studied area of interactional competence development in the professions. PMID- 17129204 TI - The dynamics of resident-patient communication: data from Canada. AB - The objectives of this study were to examine patterns of resident-patient communication and the relationship between resident patterns of speech with patient satisfaction. Forty consultations, ten in each of the four gender combinations (male resident/male patient, male resident/female patient, female resident/female patient, female resident/male patient) were audiotaped and microanalyzed using the Roter Interaction Analysis System. Several findings depart significantly from previous studies with physician-only or physician resident-mixed samples. First, the average length of the 40 consultations was 19.5 minutes, 11.3 minutes longer than consultations in a physician-only sample drawn in the same clinic previously. Second, male residents engaged in twice as much psychosocial talk as female residents and conducted longer consultations. Third, residents asked 80% of the total questions while patients asked 20% of the questions. Previous studies with physician-only or physician-resident-mixed samples reported that physicians ask 89-99% of the total questions. Finally, patients' overall satisfaction and communication satisfaction were negatively correlated with residents' positive talk, which constitutes 31% of a given resident's total utterances. In the study conducted in the same clinic with a physician-only sample, physician positive talk was 26% and physician positive talk was not correlated with patient satisfaction. Is this a signal that residents should reduce the amount of positive talk? Apparently more studies with resident-only samples are needed to answer this and other unanswered questions in the field to offer directives to resident training. PMID- 17129205 TI - Loss of self-control as excuse in group-therapy conversations for intimately violent men. AB - This article examines the constructions of loss of self-control by male perpetrators of intimate violence in therapy-group conversations. It looks at discursive strategies used by therapists and clients in therapy-group negotiations concerning the issue of self-control. The data are part of a larger corpus of videotaped and transcribed recordings of treatment groups for male perpetrators and consists of four treatment group processes. The analysis concentrates on episodes of self-control talk where a participant referred to loss of self-control when accounting for his violent behavior. The loss of self control was found to be a fundamental constituent of excuses and was used to account for the clients' behavior in past violent situations. It allowed the speaker to admit the reprehensible character of his behavior, while denying full responsibility for it. It also permitted the speaker to be portrayed as a responsible person in the present conversational group therapy context. These different representations of the past and present selves gave those presentations the appearance of inconsistency. The therapists used this inconsistency in the clients' talk to challenge the use of loss of self-control as excuse, and thus called for more responsible accounts. PMID- 17129206 TI - Integrating patients' nonmedical status in end-of-life decision making: structuring communication through 'conferencing'. AB - This paper considers the nonmedical status of patients in end-of-life decisions. Considering nonmedical factors is not yet routine, particularly in decisions to withhold or withdraw treatment. The paper advocates that non-medical factors-the capacity and willingness to withstand continuing treatment-are essential to ensure that decisions taken are in the patient's best interest. We argue that including this dimension of patient care not commonly considered gives balance to decisions about continuing treatment where its benefit is diminishing. Drawing on a qualitative study of intensive care nursing in a large public hospital in Sydney, Australia, the paper exemplifies and interprets the tendency of some clinicians to not disclose the medical and nonmedical status to conscious patients, and the environment of mistrust and conflict that can result. We propose a process of 'conferencing'-a regular, inclusive, ongoing, and dynamic process of communication begun early in the patient's admission-to allow multidisciplinary clinicians to manage their differences, agree on patient-care goals, and prepare the patient and their family for the experience of dying. By integrating both medical and nonmedical factors, conferencing becomes the means of enacting and embedding a multidisciplinary, multidimensional approach to end of-life care. PMID- 17129209 TI - Expression of VCA (viral capsid antigen) and EBNA1 (Epstein-Barr-virus-encoded nuclear antigen 1) genes of Epstein-Barr virus in Pichia pastoris and application of the products in a screening test for patients with nasopharyngeal carcinoma. AB - EBV (Epstein-Barr virus) serological tests have been used for many years as accessory diagnostic predictors of NPC (nasopharyngeal carcinoma). To date, IF (indirect immunofluorescence) assays still serve as the 'gold standard' for EBV serodiagnosis. However, IF assays are time-consuming, unsuitable for automatic handling and difficult to standardize. This makes their application in mass screening of populations inconvenient. Some of the technical difficulties associated with IF have been overcome by the development of specific ELISAs, but, at present, high sensitivity and specificity cannot be achieved simultaneously by using recombinant protein-based ELISAs, as the diagnostic value of different fragments of EBV in NPC is different. In an attempt to determine a suitable recombinant EBV protein for diagnostic purposes, fragments of EBV VCA (viral capsid antigen) and EBNA1 (Epstein-Barr-virus-encoded nuclear antigen 1) genes were expressed in the methylotrophic yeast Pichia pastoris, and a novel ELISA was established using P. pastoris-expressed VCA-BALF4 [aa (amino acids) 287-623; the BALF4 gene encodes the EBV glycoprotein gp125], EBNA1 (aa 390-641) and VCA-BFRF3 (the gene BFRF3 encodes a viral structural capsid protein or tegument protein VCA p18) proteins. Serum samples were collected from patients with NPC and healthy controls and were tested using this ELISA. The sensitivity of VCA-BFRF3, VCA BALF4 and EBNA1 tests in the NPC sera were 65.0 (195/300), 76.3 (229/300) and 81.4% (244/300) respectively, whereas the specificity of normal individuals were 92 (460/500), 96 (480/500) and 95.8% (479/500). The optimum combination is VCA BALF4 plus EBNA1, which identified 90.3% (271/300) of the NPC patients and had a specificity of 92.8% (464/500) for normal individuals. The results obtained from the evaluation of three antibodies to EBV as markers for detecting NPC suggests that a combination of EBNA1 (aa 390-641) and VCA-BALF4 (aa 287-623) assays would give better results in screening for NPC. PMID- 17129210 TI - Gender-related differences in the sympathetic vasoconstrictor drive of normal subjects. AB - The risk of cardiovascular disease has been linked to sympathetic activation and its incidence is known to be lower in women than in men. However, the effect of gender on the sympathetic vasoconstrictor drive has not yet been established. In the present study, we investigated whether there is a gender difference in MSNA (muscle sympathetic nerve activity) and blood flow, and to determine the mechanisms involved. We examined 68 normal subjects, 34 women and 34 men, matched for age, BMI (body mass index) and waist circumference. MSNA was measured as the mean frequency of single units (s-MSNA) and as multi-unit bursts (m-MSNA) from the peroneal nerve simultaneously with its supplied muscle CBF (calf blood flow). Women had lower (P=0.0007) s-MSNA (24+/-2.0 impulses/100 cardiac beats) than men (34+/-2.3 impulses/100 cardiac beats), and a greater baroreceptor reflex sensitivity controlling efferent sympathetic nerve activity than men. The sympathetic activity was inversely and directly correlated respectively, with CBF (P=0.03) and CVR (calf vascular resistance; P=0.01) in men only. The responses of an increase in CVR to cold pressor and isometric handgrip tests were significantly smaller in women (P=0.002) than in men, despite similar increases in efferent sympathetic nerve activity. Women had a lower central sympathetic neural output to the periphery, the mechanism of which involved differences in central and reflex control, as well as a lower vasoconstrictor response to this neural output. It is suggested that this may partly explain the observed lower incidence of cardiovascular events in women compared with men. PMID- 17129211 TI - Lysosomes and Fas-mediated liver cell death. AB - A number of studies, mostly performed ex vivo, suggest that lysosomes are involved in apoptosis as a result of a release of their cathepsins into the cytosol. These enzymes could then contribute to the permeabilization of the outer mitochondrial membrane; they could also activate effector caspases. The present study aims at testing whether the membrane of liver lysosomes is disrupted during Fas-mediated cell death of hepatocytes in vivo, a process implicated in several liver pathologies. Apoptosis was induced by injecting mice with aFas (anti-Fas antibody). The state of lysosomes was assessed by determining the proportion of lysosomal enzymes (beta-galactosidase, beta-glucuronidase, cathepsin C and cathepsin B) present in homogenate supernatants, devoid of intact lysosomes, and by analysing the behaviour in differential and isopycnic centrifugation of beta galactosidase. Apoptosis was monitored by measuring caspase 3 activity (DEVDase) and the release of sulfite cytochrome c reductase, an enzyme located in the mitochondrial intermembrane space. Results show that an injection of 10 microg of aFas causes a rapid and large increase in DEVDase activity and in unsedimentable sulfite cytochrome c reductase. This modifies neither the proportion of unsedimentable lysosomal enzyme in the homogenates nor the behaviour of lysosomes in centrifugation. Experiments performed with a lower dose of aFas (5 microg) indicate that unsedimentable lysosomal hydrolase activity increases in the homogenate after injection but with a marked delay with respect to the increase in DEVDase activity and in unsedimentable sulfite cytochrome c reductase. Comparative experiments ex vivo performed with Jurkat cells show an increase in unsedimentable lysosomal hydrolases, but much later than caspase 3 activation, and a release of dipeptidyl peptidase III and DEVDase into culture medium. It is proposed that the weakening of lysosomes observed after aFas treatment in vivo and ex vivo results from a necrotic process that takes place late after initiation of apoptosis. PMID- 17129212 TI - Murine [corrected] myeloid dendritic cell-dependent toll-like receptor immunity is preserved with aging. AB - The immune response is the result of the interplay between innate and adaptive immunity, yet the impact of aging on this interaction is unclear. Addressing this fundamental question will be critical for the development of effective vaccines for the rapidly rising older subpopulation that manifests increased prevalence of malignancies and infections. Therefore, we undertook the current study to investigate whether aging impairs toll-like receptor (TLR) function in myeloid dendritic cells and whether this leads to reduced T-cell priming. Our results demonstrate that innate TLR immune priming function of myeloid bone marrow derived and splenic dendritic cells (DC) is preserved with aging using both allogeneic and infectious murine experimental systems. In contrast, aging impairs in vitro and in vivo intrinsic T-cell function. Therefore, our results demonstrate that myeloid DCs manifest preserved TLR-mediated immune responses with aging. However, aging critically impairs intrinsic adaptive T-cell function. PMID- 17129213 TI - Sirtuin-independent effects of nicotinamide on lifespan extension from calorie restriction in yeast. AB - Two models have been proposed for how calorie restriction (CR) enhances replicative longevity in yeast: (i) suppression of rDNA recombination through activation of the sirtuin protein deacetylase Sir2 or (ii) decreased activity of the nutrient-responsive kinases Sch9 and TOR. We report here that CR increases lifespan independently of all Sir2-family proteins in yeast. Furthermore, we demonstrate that nicotinamide, an inhibitor of Sir2-mediated deacetylation, interferes with lifespan extension from CR, but does so independent of Sir2, Hst1, Hst2, and Hst4. We also find that 5 mm nicotinamide, a concentration sufficient to inhibit other sirtuins, does not phenocopy deletion of HST3. Thus, we propose that lifespan extension by CR is independent of sirtuins and that nicotinamide has sirtuin-independent effects on lifespan extension by CR. PMID- 17129214 TI - Disparate patterns of age-related changes in lipid peroxidation in long-lived naked mole-rats and shorter-lived mice. AB - A key tenet of the oxidative stress theory of aging is that levels of accrued oxidative damage increase with age. Differences in damage generation and accumulation therefore may underlie the natural variation in species longevity. We compared age-related profiles of whole-organism lipid peroxidation (urinary isoprostanes) and liver lipid damage (malondialdehyde) in long living naked mole rats [maximum lifespan (MLS) > 28.3 years] and shorter-living CB6F1 hybrid mice (MLS approximately 3.5 years). In addition, we compared age-associated changes in liver non-heme iron to assess how intracellular conditions, which may modulate oxidative processes, are affected by aging. Surprisingly, even at a young age, concentrations of both markers of lipid peroxidation, as well as of iron, were at least twofold (P < 0.005) greater in naked mole tats than in mice. This refutes the hypothesis that prolonged naked mole-rat longevity is due to superior protection against oxidative stress. The age-related profiles of all three parameters were distinctly species specific. Rates of lipid damage generation in mice were maintained throughout adulthood, while accrued damage in old animals was twice that of young mice. In naked mole-rats, urinary isoprostane excretion declined by half with age (P < 0.001), despite increases in tissue iron (P < 0.05). Contrary to the predictions of the oxidative stress theory, lipid damage levels did not change with age in mole-rats. These data suggest that the patterns of age-related changes in levels of markers of oxidative stress are species specific, and that the pronounced longevity of naked mole-rats is independent of oxidative stress parameters. PMID- 17129215 TI - Trade-offs between longevity and pathogen resistance in Drosophila melanogaster are mediated by NFkappaB signaling. AB - The innate immune response protects numerous organisms, including humans, from the universe of pathogenic molecules, viruses and micro-organisms. Despite its role in promoting pathogen resistance, inappropriate activation and expression of NFkappaB and other immunity-related effector molecules can lead to cancer, inflammation, and other diseases of aging. Understanding the mechanisms leading to immune system activation as well as the short- and long-term consequences of such activation on health and lifespan is therefore critical for the development of beneficial immuno-modulating and longevity-promoting interventions. Mechanisms of innate immunity are highly conserved across species, and we take advantage of genetic tools in the model organism, Drosophila melanogaster, to study the effects of acute and chronic activation of immunity pathways on pathogen resistance and general fitness of adult flies. Our findings indicate that fat body specific overexpression of a putative pathogen recognition molecule, peptidoglycan recognition protein (PGRP-LE), is sufficient for constitutive up regulation of the immune response and for enhanced pathogen resistance. Primary components of fitness are unaffected by acute activation, but chronic activation leads to an inflammatory state and reduced lifespan. These phenotypes are dependent on the NFkappaB-related transcriptional factor, Relish, and they establish a mechanistic basis for a link between immunity, inflammation, and longevity. PMID- 17129216 TI - The window and mechanisms of major age-related decline in the production of new neurons within the dentate gyrus of the hippocampus. AB - While it is well known that production of new neurons from neural stem/progenitor cells (NSC) in the dentate gyrus (DG) diminishes greatly by middle age, the phases and mechanisms of major age-related decline in DG neurogenesis are largely unknown. To address these issues, we first assessed DG neurogenesis in multiple age groups of Fischer 344 rats via quantification of doublecortin-immunopositive (DCX+) neurons and then measured the production, neuronal differentiation and initial survival of new cells in the subgranular zone (SGZ) of 4-, 12- and 24 month-old rats using four injections (one every sixth hour) of 5' bromodeoxyuridine (BrdU), and BrdU-DCX dual immunostaining. Furthermore, we quantified the numbers of proliferating cells in the SGZ of these rats using Ki67 immunostaining. Numbers of DCX+ neurons were stable at 4-7.5 months of age but decreased progressively at 7.5-9 months (41% decline), 9-10.5 months (39% decline), and 10.5-12 months (34% decline) of age. Analyses of BrdU(+) cells at 6 h after the last BrdU injection revealed a 71-78% decline in the production of new cells per day between 4-month-old rats and 12- or 24-month-old rats. Numbers of proliferating Ki67+ cells (putative NSCs) in the SGZ also exhibited similar (72-85%) decline during this period. However, the extent of both neuronal differentiation (75-81%) and initial 12-day survival (67-74%) of newly born cells was similar in all age groups. Additional analyses of dendritic growth of 12-day old neurons revealed that newly born neurons in the aging DG exhibit diminished dendritic growth compared with their age-matched counterparts in the young DG. Thus, major decreases in DG neurogenesis occur at 7.5-12 months of age in Fischer 344 rats. Decreased production of new cells due to proliferation of far fewer NSCs in the SGZ mainly underlies this decline. PMID- 17129217 TI - Gender differences in free radical homeostasis during aging: shorter-lived female C57BL6 mice have increased oxidative stress. AB - Gender is a profound determinant of aging and lifespan, but little is known about gender differences in free radical homeostasis. Free radicals are proposed as key elements in the multifactorial process of aging and it is predicted that the longer-lived gender should have lower levels of oxidative stress. While the majority of studies on aging have included a single gender, recent studies in rats compared genders and found that females, the longer-lived sex, had lower oxidative stress and mitochondrial dysfunction than males. We explored the association between oxidative stress and gender-specific aging in C57BL6 mice, in which females are the shorter-lived gender. Reactive oxygen species (ROS) were measured in young and old mice by confocal imaging of dihydroethidium (DHE) oxidation in the brain, and by electron paramagnetic resonance (EPR) spectrometry of isolated brain mitochondria. Both genders exhibited significant age-dependent increases in ROS. However, females had a greater increase with age than males in DHE oxidation but not mitochondrial EPR. Superoxide dismutase 1 (Sod1) and glutathione peroxidase 1 (GPx1) protein levels were lower in old females. To determine whether enhancing antioxidant defenses would eliminate gender differences in lifespan, mice were treated chronically with a superoxide dismutase mimetic. Treatment blocked the age-dependent increase in ROS, with a greater effect in females on DHE oxidation, but not mitochondrial EPR. Treatment also increased lifespan to a greater degree in females. Our results indicate that differences in ROS homeostasis contribute to gender divergence in survival, but also suggest that mitochondrial superoxide production may not be primarily responsible for gender differences in lifespan. PMID- 17129220 TI - Amnesia for electric dental pulp stimulation and picture recall test under different levels of propofol or midazolam sedation. AB - AIM: To compare the amnesic effect of propofol and midazolam to electric dental pulp stimulation (invasive) and picture recall test (non-invasive) at two sedation levels with the aid of bispectral index (BIS) monitoring. METHODS: The subjects were 10 male volunteers (24-34 years) classified as ASA physical status I. Propofol was administered to achieve a sedation score of three with a target controlled infusion technique; it was then regulated to give a sedation score of two (P group). Midazolam was administered by a titration dosage to achieve a sedation score of three (M group). It then gradually decreased to give a sedation score of two. The BIS score, sedation score, plasma/serum concentration of propofol and midazolam, blood pressure, pulse rate, respiratory rate, end-tidal CO(2) tension and arterial oxygen saturation were observed at each sedation level in both groups. Amnesic effects were evaluated using a picture recall test and electric dental pulp stimulation. RESULT: No difference was observed in the amnesic effect evaluated by picture recall test at the two sedation levels. Likewise, there was no difference at a sedation score of three when the amnesic effect was evaluated by electric dental pulp stimulation. In contrast, a significant difference was observed at a sedation score of two; midazolam produced amnesia in more subjects than did propofol. CONCLUSION: Propofol and midazolam did not show any significant difference in amnesic effects to non invasive stimuli. For invasive stimuli, midazolam showed a stronger amnesic effect at the moderate sedation level, but not at the deeper sedation level. PMID- 17129221 TI - Recombinant activated factor VII efficacy and safety in a model of bleeding and thrombosis in hypothermic rabbits: a blind study. AB - BACKGROUND: Recombinant activated factor VII (rFVIIa) is increasingly used to secure hemostasis in hemorrhagic situations in trauma and surgical patients. Hypothermia is often observed in these clinical settings. OBJECTIVE: To study the efficacy and safety of rFVIIa in hypothermia in a rabbit model of bleeding and thrombosis. METHODS: Sixty-nine rabbits were anesthetized, ventilated and monitored for blood pressure, temperature and carotid flow. The Folts model was used: a stenosis (75%) and an injury were carried out on the carotid artery, inducing thrombosis. Blood flow decreased as thrombus size increased until the pressure gradient was such that the thrombus was released and local arterial blood flow was suddenly restored. This is known as a cyclic flow reduction (CFR). After counting baseline CFRs during a 20-min period (P1), rabbits were randomized blindly to one of four groups: normothermic (NT) placebo or rFVIIa (150 microg kg(-1)), hypothermic (HT) (34 degrees C) placebo or rFVIIa. Then CFRs were recorded over a second period (P2). At the end of the experiment, a hepato splenic section was performed and the amount of blood loss was recorded. After each period, the following were measured: ear immersion bleeding time (BT), hemoglobin, platelet count, prothrombin time (PT), activated partial thromboplastin time (aPTT) and fibrinogen. RESULTS: Hypothermia increased BT and blood loss. These effects were reversed by rFVIIa. In NT rabbits, rFVIIa shortened BT but did not reduce blood loss. rFVIIa-treated rabbits bled similarly regardless of temperature. The incidence of CFRs was higher in treated than placebo animals regardless of temperature. rFVIIa decreased PT and aPTT without modifying platelet count or fibrinogen level. CONCLUSION: Hemostatic efficacy of rFVIIa was maintained in hypothermia. However, the number of CFRs was higher in the rFVIIa-treated group than in the placebo groups, whether for NT or HT rabbits. PMID- 17129222 TI - Various distinctive cytogenetic abnormalities in patients with acute myeloid leukaemia aged 60 years and older express adverse prognostic value: results from a prospective clinical trial. AB - Diagnostic cytogenetic abnormalities are considered important prognostic factors in patients with acute myeloid leukaemia (AML). However, the prognostic assessments have mainly been derived from patients with AML aged <60 years. Two recent studies of AML patients of 60 years and older proposed prognostic classifications with distinct discrepancies. To further study the prognostic value of cytogenetic abnormalities in this patient population, we have evaluated cytogenetic abnormalities in a series of 293 untreated patients with AML aged 60 years and older, included in a randomised phase 3 trial, also in relation to patient characteristics and clinical outcome. The most frequently observed cytogenetic abnormality was trisomy 8 (+8), in 31 (11%) patients. Abnormalities, such as -5, 5q-, abn(17p) and abn(17q), were almost exclusively present in complex karyotypes. A relatively favourable outcome was only observed in five patients with core-binding factor abnormalities t(8;21) and inv(16)/del(16)/t(16;16). However, most of the other evaluated cytogenetic abnormalities, such as 5q-, -7, +8, abn(17p), abn(17q), and complex aberrations expressed a more adverse prognosis when compared with patients with AML aged 60 years and older with a normal karyotype. Large studies to confirm the prognosis of individual cytogenetic aberrations are warranted. PMID- 17129223 TI - Minimal residual disease in chronic lymphocytic leukaemia: is it ready for primetime? AB - New therapeutic modalities have substantially improved response rates and outcomes in chronic lymphocytic leukaemia (CLL), yet the mindset remains that palliation is the only goal of therapy because the disease is considered incurable. Ultimately, all patients relapse despite achieving an initial response, as minimal residual disease (MRD) persisting after therapy eventually evolves into morphological and clinical recurrence. The emergence of immune-based combination therapies capable of inducing molecular remissions, the availability of highly sensitive assays that detect MRD, and emerging data showing a longer duration of response or longer survival in patients with no detectable disease, suggest that eradicating MRD may be a reasonable option for some patients. Moreover, novel biological prognostic markers have divided CLL into favourable and unfavourable subtypes, arguing in favour of defining different goals of therapy for different patients. Clinicians are increasingly challenged with the task of how best to incorporate MRD assessment into clinical practice, especially in an era when these novel prognostic factors exist. This review summarises the current understanding of MRD from a clinical standpoint, suggests that MRD eradication maybe a reasonable option for some patients, and argues in favour of designing large randomised studies to determine whether MRD-negative remission improves outcome. PMID- 17129225 TI - Expression and production of aberrant PAX5 with deletion of exon 8 in B-lineage acute lymphoblastic leukaemia of children. AB - Summary We investigated PAX5 expression in childhood B-lineage acute lymphoblastic leukaemia (ALL). Seven of 21 children with B-lineage ALL had multiple PAX5 variants, while 14 children and healthy controls showed full-length (FL) and one variant PAX5. By Western blotting, healthy controls displayed Pax5 FL, while one short Pax5, derived from the deletion of exon 8 (Pax5-DeltaE8) was produced in 90% of ALL samples, as well as in ALL cell lines. PAX5-DeltaE8 lacked more than 50% of the transactivation domain, indicating that aberrant Pax5 production might lead to the arrest of B-cell differentiation, contributing to the pathogenesis of B-lineage ALL. PMID- 17129226 TI - Clinical and molecular analysis of haemoglobin H disease in Sardinia: haematological, obstetric and cardiac aspects in patients with different genotypes. AB - In this study, 251 Sardinian patients (187 adults and 64 children) with haemoglobin (Hb) H disease were evaluated. Two-hundred and sixteen patients (86%) had the deletional type (- -/-alpha) and 36 (14%) patients had the non-deletional type (- -/alpha(ND)alpha). A clear genotype-phenotype correlation was found, with the non-deletional type more severe than the deletional type. Diagnosis of Hb H disease was incidental in about 60% of cases. Aplastic crises due to B19 parvovirus infection were found in five patients (2.1%), while 23 patients (9.6%) experienced one or more haemolytic crises. Nineteen patients with Hb H received sporadic red blood cell transfusions and three patients were repeatedly transfused. Forty-seven of 61 married women (77%) had 82 pregnancies. In children, mean serum ferritin was 87 +/-92 mug/l and in adults, was 192 +/- 180 mug/l in females and 363 +/- 303 mug/l in males. For the 98 male patients, a significant correlation was found between ferritin values and age (r2 = 0.33, P < 0.0001). In the Sardinian population, Hb H disease needs regular monitoring for early detection and treatment of possible complications, such as worsening of anaemia that may require red cell transfusion, cholelithiasis and iron overload. PMID- 17129227 TI - A woman with black urine. PMID- 17129228 TI - Computed tomography findings in a patient with severe graft-versus-host disease of the gut. PMID- 17129229 TI - The t(14;19)(q32;q13)-positive small B-cell leukaemia: a clinicopathologic and cytogenetic study of seven cases. AB - The t(14;19)(q32;q13), involving the BCL3 locus at chromosome 19q13 and the immunoglobulin heavy chain gene at 14q32, is a rare recurrent cytogenetic abnormality identified in B-cell neoplasms, most of which have been classified as chronic lymphocytic leukaemia (CLL) in the literature. We describe the clinicopathological, immunophenotypic and cytogenetic findings in seven patients with B-cell neoplasms associated with t(14;19)(q32;q13). There were five men and two women, with a median age of 48 years (range 33-68). All had absolute lymphocytosis, six had lymphadenopathy, and one had splenomegaly. Lymphocytes in blood and bone marrow aspirate smears were predominantly small and cytologically atypical. Flow cytometric immunophenotyping showed an atypical immunophenotype with low CLL scores. The growth pattern in bone marrow biopsy specimens was interstitial to diffuse; immunohistochemical stains were positive for bcl3 and negative for cyclin D1. Lymph node biopsy specimens of two patients revealed total architectural effacement by neoplasm with proliferation centres. In addition to t(14;19), cytogenetic studies demonstrated trisomy 12 in five patients. These results suggest that B-cell neoplasms with the t(14;19)(q32;q13) present frequently as leukaemia composed of small B-lymphocytes and share many features with CLL. However, these neoplasms also differ from CLL cytologically and in their immunophenotype. PMID- 17129230 TI - Ewing sarcoma diagnosed by fluorescence in situ hybridisation of bone marrow films. PMID- 17129231 TI - 18F-Fluorodeoxyglucose positron emission tomography for evaluation of intravascular large B-cell lymphoma. PMID- 17129232 TI - Molecular therapies in beta-thalassaemia. AB - The beta-thalassaemias have a major global impact on health and mortality. Allogeneic haemopoietic stem cell transplantation is the only approach that may lead to a cure but this approach is not available to most patients. The mainstay treatment for the majority remains life-long blood transfusion in combination with a rigorous regime of iron chelation. Improved understanding of the pathophysiology and molecular basis of the disease has provided clues for more effective strategies that aim to correct the defect in beta-globin chain synthesis at the primary level or redress the alpha/beta-globin chain imbalance at the secondary level. Improved understanding of the molecular basis of the disease complications, such as iron overloading, has also provided clues for potential molecular targets at the tertiary level. PMID- 17129233 TI - Relapsing acute myeloid leukaemia manifesting as uveitis with hypopyon. PMID- 17129234 TI - Topical eutectic mixture for premature ejaculation (TEMPE): a novel aerosol delivery form of lidocaine-prilocaine for treating premature ejaculation. AB - OBJECTIVE: To evaluate, in a phase II study, the efficacy and safety of a topical eutectic mixture for premature ejaculation (TEMPE), a metered-dose aerosol spray containing a eutectic mixture of lidocaine and prilocaine, as a treatment for PE. PATIENTS AND METHODS: Men with PE (Diagnostic and Statistical Manual-IV definition) aged 18-75 years were randomized into a double-blind, placebo controlled study in the UK and the Netherlands. Efficacy variables included the mean change in intravaginal ejaculatory latency time (IELT) from baseline and the proportion of patients who achieved an IELT of > or = 4, > or = 3 or > or = 2 min on two occasions, and the effect of TEMPE on the index of ejaculatory control (IEC) and sexual quality-of-life (SQoL) scores of patients and their partners. Safety and adverse event data were also collected. Fifty-four patients were randomized and received study treatment. RESULTS: The observed mean change in IELT from baseline to the end of the treatment period was 3.8 min in the TEMPE group and 0.7 min in the placebo group, and when adjusted for baseline and centre was 2.4 times higher in the TEMPE than the placebo group (P < 0.01). The efficacy of TEMPE in increasing IELT was further supported by positive trends in the other efficacy endpoints. The proportion of men who had an IELT time > or = 2, > or = 3 or > or = 4 min on two occasions after treatment was 11/20 (55%), 8/20 (40%) and 5/25 (20%) in the TEMPE group, and 8/23 (35%), 3/23 (13%) and 3/23 (13%) in the placebo group, respectively, although these differences were not statistically significant. Improvements in IEC and SQoL (male and female) scores also showed trends towards greater efficacy for TEMPE than placebo. In all, 35 of 42 (83%) patients considered the spray easy to use. Mild to moderate local numbness occurred in three (12%) of the TEMPE-treated patients but did not lead to discontinuation. CONCLUSION: Topical treatment with TEMPE produced a statistically and clinically significant increase in IELT compared with placebo, and resulted in positive trends in ejaculatory control and SQoL. TEMPE was considered easy to use and was well tolerated. The data support the conduct of further large-scale studies to establish the utility of TEMPE as a first-line treatment for PE. PMID- 17129257 TI - The cosmeceutical conundrum. PMID- 17129258 TI - Commentary on cellulite: skin mechanobiology and the waist-to-hip ratio. AB - Cellulite is a gender-related condition which is the clinical expression of conformational changes taking place in the fibrous strands partitioning the hypodermis. The affected skin areas are those where fat deposition is under the influence of estrogens. Some hypodermal fibrous strands become enlarged and others become loose and look similar to striae distensae. Cellulite is not a result of increased body mass, but its aspect may be influenced by the waist-to hip ratio. PMID- 17129259 TI - Capillaroscopy and videocapillaroscopy assessment of skin microcirculation: dermatologic and cosmetic approaches. AB - BACKGROUND: Different noninvasive bioengineering techniques exist to study the microvasculature of the skin and the dynamics of the microcirculation. The goal of these techniques is to visualize the skin capillary circulation easily and directly. Indeed, this information is irreplaceable to study the physiology and physiopathology of the skin capillary circulation efficiently. AIMS: Capillaroscopy and video-capillaroscopy techniques are presented with different methods to study the capillary structure of the skin. METHODS: The methods presented in this work include image processing analysis combining morphology, statistics, geometry, and neural network detection designed to quantify the microcirculation and to follow its evolution. To illustrate the combination of these techniques and methods, different examples of their application are described, in dermatology (hypertension, venous insufficiency, age-related changes) as well as in cosmetology (rosacea and erythrosis assessment). CONCLUSION: The determination of structural or dynamic changes in the cutaneous microcirculation belongs to the noninvasive techniques of the biometrological domain. Thus, every capillary modification resulting from topical cosmetic products, or chemical agents can be observed. In pathology, numerous conditions can be better examined with this system. Associated with the potential of numerical image analysis, capillaroscopy techniques will probably extend their application fields to the assessment of the influence of arterial and venous diseases on the skin nutritional circulation. PMID- 17129260 TI - Treatment of palmar hyperhidrosis with botulinum toxin type A: 44 months of experience. AB - BACKGROUND: Palmar hyperhidrosis (PH) can produce social and occupational difficulties and reduce the quality of life of those who suffer from this kind of problem. When dealing with focal hyperhidrosis, the patients' attitudes and their subjective approaches regarding the process may influence the objective evaluation of the disorder. OBJECTIVE: To evaluate, by means of a scale, the subjective improvement of sweat production after treatment with botulinum toxin type A (BTX-A) in a group of patients with severe, invalidating PH. Patients and methods Over a period of 44 months, 69 patients were treated and followed-up, 27 patients had to be treated twice, and 11 patients required a third application; 80-100 U was injected in each palm. Regional nerve block was performed before the procedure. The patients were asked to evaluate their improvement at 1, 3, 6, 9, and 12 months of baseline. RESULTS: At 1 month, 53.6% of the patients reported an excellent improvement. Three months later, results were still excellent in 33.3% of the patients, and acceptable in 29%. From then on there was a statistically significant decrease of BTX-A effectiveness. A second application was carried out at an interval of 7.5 +/- 2.6 months, and a third one at 9 +/- 4.4 months. The following complications could be observed: transitory weakness of hand muscles in 13 patients, wrist pain in 5 patients, and cramps in 1 patient. CONCLUSIONS: Botulinum toxin is an effective alternative for the treatment of severe, invalidating PH. The maximum improvement persists up to 3 months; from then on, the effects slowly diminish. In our experience, there were no statistically significant differences with further applications of BTX-A. The patients' subjective assessment can be used to evaluate the sweat production after treatment with BTX-A in those medical centers where a more objective evaluation becomes difficult or impossible. PMID- 17129261 TI - Clinical efficacy assessment in photodamaged skin of 0.5% and 1.0% idebenone. AB - Idebenone is an antioxidant lower molecular weight analogue of coenzyme Q10. Previously, idebenone was shown to be a very effective antioxidant in its ability to protect against cell damage from oxidative stress in a variety of biochemical, cell biological, and in vivo methods, including its ability to suppress sunburn cell (SBC) formation in living skin. However, no clinical studies have been previously conducted to establish the efficacy of idebenone in a topical skincare formulation for the treatment of photodamaged skin. In this nonvehicle control study, 0.5% and 1.0% idebenone commercial formulations were evaluated in a clinical trial for topical safety and efficacy in photodamaged skin. Forty-one female subjects, aged 30-65, with moderate photodamaged skin were randomized to use a blind labelled (either 0.5% or 1.0% idebenone in otherwise identical lotion bases) skincare preparation twice daily for six weeks. Blinded expert grader assessments for skin roughness/dryness, fine lines/wrinkles, and global improvement in photodamage were performed at baseline, three weeks and six weeks. Electrical conductance readings for skin surface hydration and 35 mm digital photography were made at baseline after six weeks. Punch biopsies were taken from randomly selected subjects, baseline and after six weeks, and stained for certain antibodies (interleukin IL-6, interleukin IL-1b, matrixmetalloproteinase MMP-1, collagen I) using immunofluorescence microscopy. After six weeks' use of the 1.0% idebenone formula, a 26% reduction in skin roughness/dryness was observed, a 37% increase in skin hydration, a 29% reduction in fine lines/wrinkles, and a 33% improvement in overall global assessment of photodamaged skin. For the 0.5% idebenone formulation, a 23% reduction in skin roughness/dryness was observed, a 37% increase in skin hydration, a 27% reduction in fine lines/wrinkles, and a 30% improvement in overall global assessment of photodamaged skin. The immunofluorescence staining revealed a decrease in IL-1b, IL-6, and MMP-1 and an increase in collagen I for both concentrations. PMID- 17129262 TI - Striae distensae in darker skin types: the influence of melanocyte mechanobiology. AB - INTRODUCTION: Striae distensae in White people are commonly described as being reddish at first and turning white in time. This dual perception has been challenged. Indeed, the color difference between striae distensae and their surrounding skin varies on a wide range in part depending upon typology. AIM: To study striae nigrae that are found only in subjects of darker complexion. MATERIALS AND METHODS: In the present study conducted in 44 darker skin adults, dermoscopy was combined to reflectance colorimetry. RESULTS: Striae nigrae exhibited hypermelanosis of the epidermal rete ridges crossing transversally the lesions in a laddering pattern. By contrast, striae albae showed only faint melanotic networks. CONCLUSIONS: As striae distensae are in part under the influence of skin stretching, their colors are likely to be controlled by some mechanobiological process activating or inhibiting melanogenesis in people of darker complexion. No argument involving an inflammatory mechanism is supported by the present observations. PMID- 17129263 TI - Cumulative irritation patch test of sanitary pads on sensitive skin. AB - BACKGROUND: Tools are needed to assess the effects of various products on sensitive skin. AIM: To investigate the cutaneous compatibility of various sanitary pads in people with self-declared sensitive skin. PATIENTS/METHODS: Eight subjects, who considered their skin to be sensitive and who reported adverse skin responses to everyday products or clothing, were patch tested on the arm with two, low-irritancy sanitary pads for four consecutive 24-hour periods. Test products differed only in their surface covering. Sodium lauryl sulfate solution (0.1% w/v) and physiological saline served as standard irritant and nonirritant controls, respectively. Skin irritation (erythema) was graded after each 24-hour period. RESULTS: No significant difference in skin erythema scores was observed between groups (scores on day 4: 1.06 +/- 0.11 vs. 1.25 +/- 0.13, on a five-point scale, P < 0.05). The temporal development of cumulative irritation associated with the pads was comparable to that observed with the nonirritant control (a plateau in maximal group scores) and distinct from that of the irritant control (continuously rising scores). CONCLUSION: Sanitary pads under investigation elicited negligible cumulative irritation in a four-day patch test on subjects with self-declared sensitive skin. The temporal pattern of cumulative response was consistent with the inherently low irritation potential of the products. PMID- 17129264 TI - Adverse effect of soft tissue augmentation with hyaluronic acid. AB - Cutaneous fillers have been employed in the treatment of wrinkles and scars. The high biocompatibility and rare adverse effects described in the literature represent advantages of hyaluronic acid (HA) compared to other fillers. We report the case of a patient submitted to soft tissue augmentation with HA (Restylane) for the treatment of wrinkles on the lower third of the face who presented a foreign-body type granulomatous reaction which revised the literature. PMID- 17129265 TI - Formulations of hydrogels and lipogels with vitamin E. AB - Vitamin E, a topically administered antioxidant, reduces erythema, photoaging, photocarcinogenesis, edema, and skin hypersensitivity associated with exposure to ultraviolet B (UVB) radiation. Virgin olive oil, which also has antioxidant properties, reduces the number of, and delays the onset of skin cancer induced by UVB radiation when used after sunbathing. Topical use after sunbathing of formulations that contain virgin olive oil and vitamin E may therefore reduce the number and delay the onset of UVB-related skin cancer in humans. We designed formulations of gels with olive oil (lipogels) and hydrogels containing 2% tocopherol. The formulations were optimized on the basis of rheological, organoleptic, biopharmaceutical, and cosmetic criteria. Different formulae for hydrogels with vitamin E were ideal for application after exposure to solar radiation because of their good organoleptic properties. Vitamin E lipogels are of potential use in cosmetics such as locally acting anti-aging treatments because of the antioxidant effects of vitamin E. PMID- 17129266 TI - An evaluation of topical 3% salicylic acid and 1% hydrocortisone in the maintenance of scalp pruritus. AB - BACKGROUND: Scalp pruritus is a common condition causing dermatologic distress. The presence of skin scale on clothing is cosmetically undesirable and scalp scratching in public is socially embarrassing. Scratching can also result in removal of the cuticle and premature hair shaft fracture. OBJECTIVE: To demonstrate the efficacy of 3% salicylic acid in combination with 1% hydrocortisone in the treatment of scalp pruritus. METHODS: Sixty subjects with moderate scalp scaling and scalp pruritus were enrolled in a three-arm double blind 2-week study. The 20 subjects in arm 1 applied a topical 1% hydrocortisone product twice daily. The 20 subjects in arm 2 applied a topical 3% salicylic acid product twice daily. Lastly, the 20 subjects in arm 3 applied a topical 3% salicylic acid product in the morning and a topical 1% hydrocortisone product in the evening. Evaluations were performed at baseline, after 1 week of treatment, and after 2 weeks of treatment. The study investigator evaluated the subjects for scalp scale, erythema, excoriation, and overall assessment. In addition, scalp scale scrapings were collected and analyzed to gain further insight into scalp scale morphology. Subject assessments and scalp photography was also performed. RESULTS: The investigator assessments revealed less excoriation in the hydrocortisone twice daily arm over the salicylic acid twice daily arm (P = 0.03), which might be expected because of its anti-inflammatory effect. The morning salicylic acid application and evening hydrocortisone application arm performed better than the salicylic acid twice daily group at week 2 in terms of erythema (P = 0.02), excoriation (P = 0.03), and overall assessment (P = 0.01). Scalp scale scrapings revealed the least amount of retained skin scale in the combination salicylic acid/hydrocortisone group. CONCLUSION: The combination of a 3% salicylic acid keratolytic combined with a 1% hydrocortisone anti-inflammatory provides the best relief of scalp pruritus. PMID- 17129267 TI - Liposculpture of the aging face and neck. PMID- 17129269 TI - Microliposuction and fat transfer for advanced repair of liposuction defects. PMID- 17129268 TI - Side effects of nail cosmetics. AB - Cosmetics applied on the nail encompass three types: (1) coatings that harden upon evaporation; (2) coatings that polymerize; and (3) stick-on nail dressings (synthetic covers). The adverse reactions induced by the two first types present with both local reactions and distant contact dermatitis. Whereas nail enamel applications result especially in ectopic contact dermatitis, polymerizing coatings and synthetic covers represent the main culprit for sometimes severe, local reactions. Whatever the nature of the nail cosmetics, they may produce some adverse reactions. They include: (1) local reactions to cosmetics applied on the nail; (2) distant reactions resulting from the use of nail cosmetics; (3) systemic side effects of nail cosmetics; and (4) infection risks from nail cosmetics. PMID- 17129270 TI - The color of the skin: psycho-anthropologic implications. PMID- 17129271 TI - The prevalence of chronic pain in the United States remains high, imposing a large economic burden on patients and society. PMID- 17129272 TI - Botulinum neurotoxins: origins and basic mechanisms of action. PMID- 17129274 TI - Treatment of headache pain with botulinum neurotoxins. PMID- 17129273 TI - Treatment of neck and shoulder pain with botulinum neurotoxins. PMID- 17129275 TI - Overview of pain management. PMID- 17129276 TI - Treatment of low back pain with botulinum neurotoxins. PMID- 17129277 TI - Treatment algorithm overview: BoNT therapy for pain. PMID- 17129278 TI - Botulinum neurotoxin in chronic pain management. PMID- 17129279 TI - Pain processing: paradoxes and predictions. AB - During the last 25 years, there have been substantial advances in our understanding of the physiology and pathophysiology of pain. The development of animal models that more closely mimic clinical pain in humans has helped elucidate the putative mechanisms by which chronic pain develops and is maintained. However, our increased understanding of the neurobiology of pain has not translated into breakthrough treatments for pain management. As such, chronic pain is still primarily managed by drugs whose primary indication does not include pain (eg, antidepressants, anticonvulsants, antiarrhythmics, local anesthetics). These adjuvant analgesics have come into favor despite the fact that the mechanisms through which these drugs provide pain relief remain either largely unknown or are not selective for a single target. Moreover, the efficacy of adjuvant analgesics in animal models of pain is often validated only after case studies or clinical trials have been reported. This retrospective validation of "novel" analgesics in animal models of pain raises a question of the predictive validity of these models. This article reviews the use of several adjuvant and standard analgesics currently used to treat difficult-to-manage pain. What can these drugs teach us about the development of novel pain medicines? Within this context, the use of animal models of pain to predict analgesic efficacy in clinical pain conditions is considered. PMID- 17129280 TI - Epidemiology, pathophysiology, and management of complex regional pain syndrome. AB - Complex regional pain syndromes (CRPS) are challenging neuropathic pain states quite difficult to comprehend and treat. Although not yet fully understood, advances are being made in the knowledge of the mechanisms involved with CRPS. Patients often present with incapacitating pain and loss of function. Patients suffering from these disorders need to have treatment plans tailored to their individual problems. A comprehensive diagnostic evaluation and early and aggressive therapeutic interventions are imperative. The therapeutic approach often calls for a combination of treatments. Medications such as antiepileptics, opioids, antidepressants, and topical agents along with a rehabilitation medicine program can help a major portion of patients suffering from these disorders. Implantable devices can aid those patients with CRPS. While progress is being made in treating patients with CRPS, it is important to remember that the goals of care are always to: 1) perform a comprehensive diagnostic evaluation, 2) be prompt and aggressive in treatment interventions, 3) assess and reassess the patient's clinical and psychological status, 4) be consistently supportive, and 5) strive for the maximal amount of pain relief and functional improvement. In this review article, the current knowledge of the epidemiology, pathophysiology, diagnostic, and treatment methodologies of CRPS are discussed to provide the pain practitioner with essential and up-to-date guidelines for the management of CRPS. PMID- 17129281 TI - Differential diagnosis of local cervical syndrome versus cervical brachial syndrome. PMID- 17129282 TI - Spinal cord stimulation: a comparison of efficacy versus other novel treatments for refractory angina pectoris. AB - Recently much attention has been directed toward novel treatment alternatives for refractory angina pectoris. Refractory angina is persistent stable class III or IV angina despite maximally tolerated medical treatment in patients with end stage coronary artery disease. Transmyocardial laser revascularization (TMLR), gene therapy, intermittent urokinase therapy, enhanced external balloon counterpulsation, and spinal cord stimulation have all been employed to treat refractory angina pectoris. TMLR and gene therapy are invasive open-chest procedures that have yielded controversial results. Intermittent urokinase and enhanced external balloon counterpulsation studies have limited follow-up times and require multiple clinic visits for treatment. Spinal cord stimulation has a proven short- and long-term efficacy and cost-effectiveness in the treatment of refractory angina. When compared to coronary artery bypass grafting (CABG), it has been shown to decrease the frequency of anginal attacks and consumption of short-acting nitrates to the same extent in refractory angina. Spinal cord stimulation's safety profile has also been well established and it can be used concurrently with cardiac pacemakers or MRI systems, provided the proper precautions are taken. Since spinal cord stimulation is a minimally invasive procedure with a favorable efficacy and safety profile, it should be considered as a valid treatment alternative after medical management has failed in refractory angina prior to implementing invasive modalities such as TMLR or gene therapy. PMID- 17129283 TI - Vertebroplasty. AB - Vertebroplasty is the percutaneous placement of polymethylmethacrylate (PMMA) into vertebral compression fractures for relief of pain. Polymethylmethacrylate is the cement used by orthopedic surgeons for rapid stable fixation of prosthetics in living bone. While the exact mechanism of pain relief is unknown, it is believed that the delivery of the PMMA into the fracture stabilizes the vertebral body, obtaining an analgesic effect. Vertebroplasty is an outpatient procedure that is performed with the aid of fluoroscopy. It has a high benefit/risk ratio with high success rates in comparison to extremely low complication rates. These patients consist of elderly osteoporotic patients that often times have underlying medical conditions or younger patients suffering from steroid or metastatic induced compression fractures. These patients are considered to be poor surgical candidates. In the past, this patient population has been relegated to epidural steroid injections, epidural catheters, or time contingent narcotics to control the pain. These conservative measures often lead to a patient with decreased activities of daily living and uncontrolled pain. Vertebroplasty provides a safe procedure that allows for long-term pain relief, decreased use of medication, and increased activities of daily living. This article is a review of the history, indications, contraindications, and key outcome studies. The technique is described along with complications, preprocedural care, and postprocedural care. PMID- 17129284 TI - Painless electrodiagnostic current perception threshold and pain tolerance threshold values in CRPS subjects and healthy controls: a multicenter study. AB - The purpose of this study is to evaluate both painless and painful sensory transmission in patients with Complex Regional Pain Syndrome (CRPS) using the automated electrodiagnostic sensory Nerve Conduction Threshold (sNCT) test. This test generates reliable, painless Current Perception Threshold (CPT) and atraumatic Pain Tolerance Threshold (PTT) measures. Standardized CPT and PTT measures using constant alternating current sinusoid waveform stimulus at 3 different frequencies 5 Hz, 250 Hz, and 2 kHz (Neurometer CPT/C Neurotron, Inc. Baltimore, MD) were obtained from CRPS subjects at a distal phalange of the affected extremity and at an ipsilateral asymptomatic control site. Matched sites were tested on healthy subjects. Detection sensitivities for an abnormal PTT and CPT test were calculated based on specificity of 90% as determined from data obtained from healthy controls. A Spearman rank correlation was used to test for a significant association between presence of allodynia and an abnormal PTT or CPT at any frequency tested. Thirty-six CRPS subjects and 57 healthy controls were tested. The highest detection sensitivity of the PTT test from symptomatic test sites was 63% for the finger and 71% for the toe. PTT abnormalities were also detected, to a lesser degree, at the asymptomatic control site (41% finger control site, 16% toe control site). The highest CPT detection sensitivity at the symptomatic site was 37% for the finger site and 53% for the toe site. CPT abnormalities were also detected at the asymptomatic control site (29% finger control site, 37% toe control site). Eighty-six percent of the CRPS subjects had either a PTT or CPT abnormality at any frequency at the symptomatic site. There was a significant correlation between presence of allodynia and presence of an abnormal CPT and PTT, respectively (P < .01). The correlation coefficient was lower for CPT than for PTT, ie, 0.34 versus 0.6 for the finger and 0.48 versus 0.67 for the toe, respectively. In studied CRPS patients an abnormal PTT was detected with higher sensitivity than an abnormal CPT. Assessing PTT may become a useful electrodiagnostic quantitative sensory test for diagnosing and following the course of neuropathic pain conditions. PMID- 17129285 TI - Intrathecal granuloma in patients receiving high-dose intrathecal morphine therapy: a report of two cases. PMID- 17129286 TI - Rationale and technique for single and multiple drug combinations in long-term intrathecal infusions. PMID- 17129289 TI - Forensic issues in pain: review of current practice. AB - Forensic activity in pain practice is reviewed with reference to the differing roles of the pain clinician and the independent expert. Ethical guidelines and recommendations for assessment, documentation, record review, and court testimony are discussed. Specific issues include the assessment of disability and impairment, malingering, and application of the Daubert standard in forensic pain practice. Examples of case law are reviewed for civil liability and CRPS, malpractice with opioid prescribing, and practice issues in a correctional setting. PMID- 17129290 TI - Disc related and non-disc related disorders of the thoracic spine. AB - Different anatomical structures and pathophysiological functions can be responsible for lumbar pain, each producing a distinctive clinical profile. Pain can arise from the intervertebral disc, either acutely as a primary disc related disorder, or as result of the degradation associated with chronic internal disc disruption. In either case, greatest pain provocation will be associated with movements and functions in the sagittal plane. Lumbar pain can also arise from afflictions within the zygapophyseal joint mechanism, as a result of synovitis or chondropathy. Either of these conditions will produce the greatest pain provocation during three-dimensional movements, due to maximal stress to either the synovium or joint cartilage. Finally, patients can experience different symptoms associated with irritation to the dural sleeve, dorsal root ganglion, or chemically irritated lumbar nerve root. Differential diagnosis of these conditions requires a thorough examination and provides information that can assist the clinician in selecting appropriate management strategies. PMID- 17129291 TI - Pain and quality of life. AB - This paper deals with the impact of pain on quality of life (QOL). Two major factors have contributed to the enhanced importance of QOL in recent years: the increasing frequency of pain and the resources devoted to its treatment, and the growing theoretical insight that pain affects the person as a whole. QOL is defined as the person's evaluation of his or her well-being and functioning in different life domains. It is a subjective, phenomenological, multidimensional, dynamic, evaluative, and yet quantifiable, construct. Commonly used scales for its assessment (eg, WHOQOL, SF-36) are described. Studies show that pain affects most domains of QOL, primarily physical and emotional functioning. The effect depends on the extent, duration, acuteness, intensity, affectivity, and meaning of the pain as well as on the underlying disease and the individual's characteristics. QOL is sensitive also to the treatment of pain and treatment modalities, as shown particularly by studies on cancer pain. Pain reduction is not always attended by the expected improvement in QOL. Pain is not synonymous with poor QOL and constitutes only one important factor determining QOL. The main conclusions are that treatment of pain should be multidisciplinary, considering the impact of pain and the treatment on QOL and targetting also improvement of the affected domains of QOL. PMID- 17129292 TI - Blockade of the superior hypogastric plexus block for visceral pelvic pain. AB - Visceral pelvic pain is a common problem with variable etiology. The sympathetic nervous system plays an important role in the transmission of visceral pain independent of its etiology. Five major pathways by which pelvic pain is transmitted can be identified. One of them, the superior hypogastric plexus, an extension of the preaortic plexus, is easily assessable to blockade by local anesthetics and neurolytic agents. Several techniques have been described. Long lasting pain relief with this procedure has been achieved in patients with pelvic cancer pain. However, there is a discrepancy between diagnostic and therapeutic blockade in patients with nonmalignant pain. Because a diagnostic blockade can give significant pain relief in a large variety of patients, it is worthwhile to investigate new methods that provide lasting neural blockade of the superior hypogastric plexus and long-lasting relief of this devastating condition. PMID- 17129293 TI - Neurolysis of the trigeminal and sphenopalatine ganglions. AB - Facial pain of trigeminal and sphenopalatine ganglion origin is the bain of existence for thousands of people. Treatment protocols typically begin with oral medication, usually anticonvulsants, and may progress to percutaneous and open surgical procedures. Several new medications show promise as alternatives to carbamazepine, which has been the standard first-line treatment (trigeminal neuralgia), while electromagnetic pulsed radiofrequency and gamma knife surgery are new options when the surgical route is warranted. This article will examine the anatomy of the trigeminal and sphenopalatine ganglions. Indications for neurolysis and neurolytic options will be discussed. Efficacy of the various neurolytic techniques will be thoroughly reviewed. PMID- 17129294 TI - Management of an epidural abscess after continuous epidural catheter infusion. AB - OBJECTIVE: To increase awareness of the possibility of epidural infection after continuous epidural infusion. Outline the salient diagnostic features of epidural infection. Outline a strategy to manage epidural infection and minimize morbidity. SETTING: Academic multidisciplinary pain clinic. PATIENT: A patient with a left knee meniscal tear with a history of Chronic Regional Pain Syndrome Type I (CRPS I) of the left foot. INTERVENTIONS: Attempted control of CRPS I symptoms with a tunnelled epidural catheter infusion. RESULTS AND CONCLUSIONS: The patient developed an epidural abscess diagnosed on the 11th postoperative day. The catheter was removed and the patient was treated successfully with intravenous antibiotics. PMID- 17129295 TI - Pelvic pain. PMID- 17129297 TI - Pain medicine: a medical specialty? PMID- 17129298 TI - NO pain: potential roles of nitric oxide in neuropathic pain. AB - The disabling human syndrome of "neuropathic pain" is an intractable complication of peripheral nerve injury or degeneration. A complex interaction between injured peripheral axons, sensory neurons and central nervous system signaling is thought to account for it. In this brief review, we present evidence that the free radical signaling molecule, nitric oxide (NO) may act at several levels of the nervous system during the development of experimental neuropathic pain. For example, NO may directly influence injured axons in the periphery, may indirectly influence pain by its role in the process of Wallerian degeneration, and may signal in the dorsal horn of the spinal cord. While it is premature to argue for therapeutic approaches that alter NO actions, it may be an important player in the cascade of events that generate neuropathic pain. PMID- 17129299 TI - Percutaneous lumbar discectomy: clinical response in an initial cohort of fifty consecutive patients with chronic radicular pain. PMID- 17129300 TI - Opioid pharmacotherapy in terminal disease. PMID- 17129301 TI - Xanthosis in the spinal epidural space--an epiduroscopy finding. AB - We report the presence of yellow pigment (xanthosis) in the eqidural space of a patient with a chronic painful radiculopathy. A 55-year-old Caucasian male patient with a left L5, S1 radiculopathy underwent epiduroscopy and epidural neurolysis. Epiduroscopy findings in the area of the nerve root associated with the radiculopathy included the presence of diffuse perivascular yellow pigments, increased vascularity and diffuse increased presence of sheets of fibrous tissue. This is a novo epiduroscopy finding. The case demonstrates the potential for epiduroscopy to expand our knowledge about pathological changes associated with some chronic painful conditions. PMID- 17129302 TI - Ergonomic pain--part 1: etiology, epidemiology, and prevention. AB - Work-related musculoskeletal disorders (MSDs) have reached a costly epidemic proportion in recent years, producing ergonomic pain as their most frequent clinical consequence. While work-related MSDs have declined in incidence, their prevalence continues. Individuals develop symptoms as a consequence of numerous factors that include force, sustained posture, repetitive motion, and vibration. Different combinations of these factors lend to different pathomechanical and pathophysiological consequences that appear to be unique to different regions of the body and related to distinctive work environmental and task characteristics. Federal and state agencies have made considerable attempts to regulate the work environment in a preventative fashion in order to reduce the incidence of ergonomic pain and other sequelae of work-related MSDs. PMID- 17129303 TI - The meaning response: thinking about placebos. PMID- 17129304 TI - Preemptive analgesia in laparoscopic cholecystectomy: a randomized controlled study. AB - In pain control after laparoscopic cholecystectomy, subhepatic administration of bupivacaine immediately after the creation of pneumoperitoneum has been shown to be more effective than administration before the withdrawal of the trocars. We aimed to investigate the effect of intraperitoneal bupivacaine administration to the subhepatic area before the creation of the pneumoperitoneum. Eighty patients undergoing elective laparoscopic cholecystectomy under general anesthesia were included in a prospective, randomized study. Patients received 20 mL of 0.5% bupivacaine in the subhepatic area just after intubation, before pneumoperitoneum (group 1), immediately after the creation of the pneumoperitoneum (group 2), just before the removal of the trocars (group 3), or received no local anesthetic (group 4). The degree of the postoperative pain was assessed at 0, 4, 8, 12, and 24 hours after the surgery. The consumption of analgesics (diclofenac sodium) was also recorded. The pain scores and analgesic consumption did not differ among groups 1, 3, and 4. The pain scores of group 2 were lower at each time point compared to the other groups (P < 0.001). Postoperative analgesic consumption in group 2 was reduced compared to the other groups (23.4 +/- 35.9 mg vs. 80.0 +/- 66.3 mg, P = 0.005 [group 1], 69.6 +/- 62.2 mg, P = 0.026 [group 3], and 70.0 +/- 59.9 mg, P = 0.022 [group 4]). The subhepatic infiltration of 20 mL of 0.5% bupivacaine offers good postoperative analgesia when applied just after the creation of the pneumoperitoneum, not before the pneumoperitoneum or after the termination of the pneumoperitoneum. PMID- 17129305 TI - Emotional distress and medication use in two acute pain populations: jaw and low back. AB - The present study was undertaken to compare emotional distress and functional ability between two common pain populations--acute jaw pain (JAW; n = 135) and acute low back pain (LB; n = 71). Patient groups were evaluated and compared on a variety of biopsychosocial measures, including the Beck Depression Inventory (BDI), Multidimensional Pain Inventory (MPI), Characteristic Pain Intensity (CPI), and Ways of Coping Questionnaire. Specific diagnoses were assessed using the Structured Clinical Interview of the Diagnostic and Statistical Manual (DSM IV)--I and II, and rates of Axis I and II diagnoses in these groups were further compared to base rates in the general population. Additionally, medication usage was evaluated to determine group differences. Results revealed that JAW patients had lower BDI and CPI scores, as well as a higher level of functioning on the Global Assessment of Functioning assessed by the DSM-IV. Both acute pain groups also had significantly more Axis I and II disorders than the general population. Additionally, it was found that the JAW group used more benzodiazepines, while the LB group used more schedule II narcotics. A logistic regression model created from these variables found a six-factor model, composed of the CPI, MPI coping style anomalous, Ways of Coping problem-solving, Global Assessment of Functioning, anxiety disorders, and Cluster C personality disorder diagnoses, that differentiated the JAW from the LB group. Overall, these findings suggest that the differences identified between these two groups should be considered in developing tailored treatments for individuals with acute low back and jaw pain. PMID- 17129306 TI - A study of AVINZA (morphine sulfate extended-release capsules) for chronic moderate-to-severe noncancer pain conducted under real-world treatment conditions -the ACCPT Study. AB - This study evaluated the clinical effects and pattern of use of AVINZA((r)), morphine sulfate extended-release tablets, under real-world treatment conditions. Opioid-naive subjects or subjects who have failed other opioids were eligible if they had chronic moderate-to-severe noncancer pain with an average pain score > or =4 (0-10 scale) in the preceding month. Subjects answered in-depth monthly questionnaires in three months. For the 491 evaluable subjects enrolled, the median AVINZA dose was 30 mg at baseline, titrated to 60 mg by month 1, and remained at 60 mg through month 3. Adherence was high, with almost 90% of the subjects reporting never having forgotten to take AVINZA. Mean daily pain scores (scale 0-10) significantly improved from 7.83 at enrollment to 5.77 at month 1 (P < 0.01) and then remained at this level through month 3. Significant improvements were seen in all sleep measures, and the mean Composite Sleep Score, a global measure of sleep quality (scale 0-10), significantly improved from 5.73 at baseline to 4.96 at month 3 (P < 0.01). Physical functioning was improved for activities requiring a moderate effort (P = 0.053), such as climbing one flight of stairs (P = 0.008). Two hospitalizations for nausea and vomiting were the only reported drug-related serious adverse events. This study showed that once-daily AVINZA significantly reduced pain scores, and resulted in improved sleep and physical functioning in patients with chronic moderate-to-severe pain. These results were achieved with a stable daily morphine dose over the three-month study period. PMID- 17129307 TI - Prevalence of contraindicated medical conditions and use of precluded medications in patients with painful neuropathic disorders prescribed amitriptyline. AB - Amitriptyline is a tricyclic antidepressant that is historically indicated and used to manage depression. More recently, due to clinical evidence demonstrating efficacy, it is often prescribed in the management of painful neuropathic disorders (PNDs). However, the amitriptyline label contains numerous preclusions (contraindications, warnings/precautions, drug interactions). Our objective was to measure the frequency of amitriptyline prescriptions in PND patients using the U.K. General Practice Research Database and assess whether any prescriptions were given to patients with preclusions listed in the product label. We identified a total of 13,546 patients (mean age 59 +/- 16.2 years; 66.7% female) who had a diagnosis of a PND and received > or =1 prescription for amitriptyline between July 1998 and June 2001. Nearly half (46.7%) of PND patients prescribed amitriptyline had > or =1 preclusion for its use; 3.5% had > or =1 contraindication; 22% had > or =1 warning/precaution; and 33% received > or =1 medication with a potential for drug interactions with amitriptyline. Preclusions were more likely in women than in men (48.3% vs. 43.4%, P < 0.0001); their incidence increased with age (42.8%, 50.4%, 55.1%, and 52.3% among those ages <65, 65-74, 75-84, and 85+ years, P < 0.0001), and the number of patients with preclusions was the highest in the phantom limb pain group (67.4%) and lowest in the atypical facial pain group (42.9%), P < 0.001. The average daily amitriptyline doses (starting: 33.6 +/- 32.4 mg; maintenance: 42.1 +/- 39.9 mg) were low compared to those used for the treatment of depression. Results indicate that, in a significant number of cases, the existence of preclusions did not prevent the prescribing of amitriptyline. Our findings raise a potential concern about the way this medication is being used. However, the clinical significance of these data is, as yet, unclear. Although, in theory, adverse outcomes may have been associated with this practice, we could not confirm this with this database analysis. PMID- 17129308 TI - Evaluation of aromatherapy in treating postoperative pain: pilot study. AB - This study compared the analgesic efficacy of postoperative lavender oil aromatherapy in 50 patients undergoing breast biopsy surgery. Twenty-five patients received supplemental oxygen through a face mask with two drops of 2% lavender oil postoperatively. The remainder of the patients received supplemental oxygen through a face mask with no lavender oil. Outcome variables included pain scores (a numeric rating scale from 0 to 10) at 5, 30, and 60 minutes postoperatively, narcotic requirements in the postanesthesia care unit (PACU), patient satisfaction with pain control, as well as time to discharge from the PACU. There were no significant differences in narcotic requirements and recovery room discharge times between the two groups. Postoperative lavender oil aromatherapy did not significantly affect pain scores. However, patients in the lavender group reported a higher satisfaction rate with pain control than patients in the control group (P = 0.0001). PMID- 17129309 TI - Repetitive occipital nerve blockade for cervicogenic headache: expanded case report of 47 adults. AB - PURPOSE: To evaluate the repetitive occipital nerve blocks using a nerve stimulator in the treatment of cervicogenic headache. METHODS: This prospective noncomparative clinical interventional case-series study included 47 patients suffering from cervicogenic headache using a repetitive guided occipital nerve blockade. RESULTS: Forty-one patients (87%) required more than one injection to achieve six-month pain-relief period. For every three years of headache history, the outcomes demonstrated that a patient needed one additional injection to the basic injection. CONCLUSION: The repeated nerve stimulator-guided occipital nerve blockade is a treatment mode that may relieve cervicogenic headache with no recurrence for at least six months in addition to alleviation of associated symptoms. PMID- 17129310 TI - Management of cerebral spinal fluid leak complicating spinal cord stimulator implantation. AB - Accidental puncture of the dura mater with resultant leakage of cerebral spinal fluid (CSF) and development of postdural puncture headache (PDPH) is a known potential complication of percutaneous placement of spinal cord stimulator (SCS) leads. However, the implications and management strategies for this complication have not been thoroughly reported. We report two cases of SCS lead placement complicated by CSF leak and PDPH. PMID- 17129311 TI - Morphine pump catheter rupture after percutaneous lithotripsy. PMID- 17129312 TI - Recent advances in rice biotechnology--towards genetically superior transgenic rice. AB - Rice biotechnology has made rapid advances since the first transgenic rice plants were produced 15 years ago. Over the past decade, this progress has resulted in the development of high frequency, routine and reproducible genetic transformation protocols for rice. This technology has been applied to produce rice plants that withstand several abiotic stresses, as well as to gain tolerance against various pests and diseases. In addition, quality improving and increased nutritional value traits have also been introduced into rice. Most of these gains were not possible through conventional breeding technologies. Transgenic rice system has been used to understand the process of transformation itself, the integration pattern of transgene as well as to modulate gene expression. Field trials of transgenic rice, especially insect-resistant rice, have recently been performed and several other studies that are prerequisite for safe release of transgenic crops have been initiated. New molecular improvisations such as inducible expression of transgene and selectable marker-free technology will help in producing superior transgenic product. It is also a step towards alleviating public concerns relating to issues of transgenic technology and to gain regulatory approval. Knowledge gained from rice can also be applied to improve other cereals. The completion of the rice genome sequencing together with a rich collection of full-length cDNA resources has opened up a plethora of opportunities, paving the way to integrate data from the large-scale projects to solve specific biological problems. PMID- 17129313 TI - Production of biologically active human interleukin-4 in transgenic tobacco and potato. AB - Interleukin-4 (IL-4) is a pleiotropic cytokine that plays a key regulatory role in the immune system. Recombinant human IL-4 (rhIL-4) offers great potential for the treatment of cancer, viral and autoimmune diseases. Unfortunately, the high production cost of IL-4 associated with conventional expression systems has, until now, limited broader clinical testing, particularly with regard to the more convenient and safer oral delivery of IL-4 as opposed to parenteral injection in patients. In this study, we investigated the feasibility of transgenic plants for the cost-effective production of rhIL-4. IL-4 expression vectors with different modifications under the control of a constitutive cauliflower mosaic virus 35S (CaMV 35S) promoter were introduced into tobacco by Agrobacterium-mediated transformation. Transgenic tobaccos expressing various levels of rhIL-4 protein were generated. Higher expression was achieved through IL-4 retention in the endoplasmic reticulum (ER), with the maximal accumulation being approximately 0.1% of total soluble protein (TSP) in the leaves. No improvement in expression was further achieved by replacing the native signal peptide of IL-4 with the plant signal peptide. The best rhIL-4-expressing vector shown in tobacco was selected and further transferred into potato plants. The analysis of transgenic tubers also revealed various levels of rhIL-4, with the highest being 0.08% of TSP. Sensitive in vitro T-cell proliferation assays showed that plant-derived rhIL-4 retained full biological activity. These results suggest that plants can be used to produce biologically active rhIL-4 and probably many other mammalian proteins of medical significance. Moreover, the production of plants expressing rhIL-4 will enable the testing of plant rhIL-4 by oral delivery for the treatment of clinical diseases. PMID- 17129314 TI - A comprehensive study of the use of a homologous promoter in antisense cotton lines exhibiting a high seed oleic acid phenotype. AB - As opposed to first-generation biotechnology products, such as pest-resistant crops and herbicide-resistant crops, second-generation products often utilize plant-derived, homologous or heterologous genes and/or promoters. In this study, we evaluated the ability of a promoter from a gene encoding a major storage protein in cottonseed to drive an antisense sequence of the cotton FAD2 gene to down-regulate the activity of Delta-12 desaturase enzyme in cottonseeds. The oleic acid level in the transgenic cottonseeds approximately doubled from the wild-type level of 15%, with a concomitant decrease in the level of linoleic acid. A more extensive study of one line revealed a higher degree of seed-to-seed variability in the transgenic phenotype. A thorough investigation was conducted to determine the impact of the use of a homologous promoter to drive a transgene on the activity of the endogenous promoter. The results showed that the use of the homologous alpha-globulin B promoter for transgenic purposes did not adversely affect the expression of alpha-globulin B storage protein in cottonseed. The results obtained in this investigation on the use of a homologous promoter and antisense technology will be useful in the design of strategies to alter biosynthetic pathways for nutritional quality improvements and for the production of heterologous proteins of commercial value in seeds. PMID- 17129315 TI - Enhanced recovery of a secreted recombinant human growth factor using stabilizing additives and by co-expression of human serum albumin in the moss Physcomitrella patens. AB - The production of pharmaceutical proteins in plants provides a valuable alternative to other traditional eukaryotic expression systems from economic and safety perspectives. The moss Physcomitrella patens allows the expression and secretion of complex target proteins into a simple aqueous maintenance medium, which facilitates downstream processing by rendering it less complex. To address the question of whether the addition of protein-stabilizing substances enhances the recovery of a target protein secreted into the culture medium, several additives at different concentrations were tested in a small-scale screening system. Although polyvinylpyrrolidone (PVP) and human serum albumin (HSA) showed a significant impact on protein levels, supplementation of the medium with these substances was accompanied by certain limitations in upstream processes, such as foam formation (HSA), and in downstream processes, such as reduced binding efficiency on chromatography columns (PVP), respectively. In order to reap the benefit of the enhancing effect and to avoid the given negative aspects, we developed a new strategy based on the recombinant expression of HSA in plants that are already capable of expressing a target protein. First, we analysed the expression and secretion of recombinant HSA in transiently and stably transformed wild-type (WT) plants. HSA was then co-expressed in Physcomitrella plants transgenic for human vascular endothelial growth factor (VEGF). Even with high expression levels of recombinant human VEGF (rhVEGF), the co-expression of recombinant HSA (rHSA) resulted in 48%-102% higher recovery of the target protein without concomitant negative effects on the upstream process. This strategy enables the enhanced recovery of target protein and does not require the addition of foreign components directly to the culture medium. PMID- 17129316 TI - Mutan produced in potato amyloplasts adheres to starch granules. AB - Production of water-insoluble mutan polymers in Kardal potato tubers was investigated after expression of a full-length (GtfI) and a truncated mutansucrase gene referred to as GtfICAT (GtfI without glucan-binding domain) from Streptococcus downei. Subsequent effects on starch biosynthesis at the molecular and biochemical levels were studied. Expression of the GtfICAT gene resulted in the adhesion of mutan material on starch granules, which stained red with erythrosine, and which was hydrolysed by exo-mutanase. In addition, GtfICAT expressing plants exhibited a severely altered tuber phenotype and starch granule morphology in comparison to those expressing the full-length GtfI gene. In spite of that, no structural changes at the starch level were observed. Expression levels of the sucrose-regulated, AGPase and GBSSI genes were down-regulated in only the GTFICAT transformants, showing that GtfICAT expression interfered with the starch biosynthetic pathway. In accordance with the down-regulated AGPase gene, a lower starch content was observed in the GTFICAT transformants. Finally, the rheological properties of the GTFICAT starches were modified; they showed a higher retrogradation during cooling of the starch paste. PMID- 17129317 TI - A new metabonomic strategy for analysing the growth process of the poplar tree. AB - High-resolution, magic angle spinning, proton nuclear magnetic resonance (1H HR/MAS NMR) spectroscopy and multivariate data analysis using batch processing (BP) were applied to the analysis of two different genotypes of poplar tree (Populus tremula L. x tremuloides Michx.) containing an antisense construct of PttMYB76 and control (wild-type). A gene encoding a MYB transcription factor, with unknown function, PttMYB76, was selected from a cambial expressed sequence tag (EST) library of poplar tree (Populus tremula L. x tremuloides Michx.) for metabonomic characterization. The PttMYB76 gene is believed to affect different paths in the phenyl propanoid synthetic pathway. This pathway leads to the formation of S- and G-lignin, flavonoids and sinapate esters. Milled poplar samples collected at the internodes of the tree were analysed using 1H HR/MAS NMR spectroscopy. The application of multivariate BP of the NMR results revealed a growth-related gradient in the plant internode direction, as well as the discrimination between the trees with down-regulated PttMYB76 expression and wild type populations. This paper focuses on the potential of a new analytical multivariate approach for analysing time-related plant metabonomic data. The techniques used could, with the aid of suitable model compounds, be of high relevance to the detection and understanding of the different lignification processes within the two types of poplar tree. Additionally, the findings highlight the importance of applying robust and organized multivariate data analysis approaches to facilitate the modelling and interpretation of complex biological data sets. PMID- 17129318 TI - The gene for fragrance in rice. AB - The flavour or fragrance of basmati and jasmine rice is associated with the presence of 2-acetyl-1-pyrroline. A recessive gene (fgr) on chromosome 8 of rice has been linked to this important trait. Here, we show that a gene with homology to the gene that encodes betaine aldehyde dehydrogenase (BAD) has significant polymorphisms in the coding region of fragrant genotypes relative to non-fragrant genotypes. The accumulation of 2-acetyl-1-pyrroline in fragrant rice genotypes may be explained by the presence of mutations resulting in a loss of function of the fgr gene product. The allele in fragrant genotypes has a mutation introducing a stop codon upstream of key amino acid sequences conserved in other BADs. The fgr gene corresponds to the gene encoding BAD2 in rice, while BAD1 is encoded by a gene on chromosome 4. BAD has been linked to stress tolerance in plants. However, the apparent loss of function of BAD2 does not seem to limit the growth of fragrant rice genotypes. Fragrance in domesticated rice has apparently originated from a common ancestor and may have evolved in a genetically isolated population, or may be the outcome of a separate domestication event. This is an example of effective human selection for a recessive trait during domestication. PMID- 17129319 TI - Sparking a change to evidence-based practice in health care organizations. PMID- 17129320 TI - Evidence-based practice in Australia: an unremitting challenge for quality! PMID- 17129321 TI - Thoughts on evidence-based nursing and its science--a Canadian perspective. PMID- 17129322 TI - Evidence, research, knowledge: a call for conceptual clarity. AB - OBJECTIVE: To dispel some of the conceptual confusion in the field of evidence based practice that has resulted from the overlapping use of the terms research, evidence, and knowledge. APPROACH: Theoretical discussion. FINDINGS: Often the terms research and knowledge are used as synonyms for evidence, but the overlap is never complete. The term evidence has long been understood to mean the findings of research. DISCUSSION: Recent attempts to broaden the definition of evidence to include clinical experience and experiential knowledge have been misguided. Broadening our understanding of the basis for clinical decision making and conceptualizing evidence are quite different tasks. Other factors (not other forms of evidence) do shape the clinical decision-making process, but they are not evidence. We might better term them knowledge. Confusing evidence with these other factors has hindered research and the improvement of clinical decision making in health care. We argue that this confusion results from the use of the term evidence when we really mean either research findings or knowledge. CONCLUSIONS: In this article, we have argued for specificity in the use of the term evidence. We urge the restriction of the term evidence to research findings, and while we acknowledge the importance of other influences on the clinical decision-making process, we insist that they are not evidence. The time has come to value personal experience and experiential knowledge for what they are-we should not have to disguise them as types of evidence for them to be deemed of any value. Being specific to language, the goal is to improve clinical decision making by increasing practitioners' reliance on research findings (evidence) while acknowledging (and valuing) the important part played by other forms of knowledge in the decision-making process. The distinctions are important. PMID- 17129324 TI - The effectiveness of intervention studies to decrease alcohol use in college undergraduate students: an integrative analysis. AB - AIMS: This analysis was performed to critique intervention studies targeted at decreasing alcohol use in college students for the purpose of (1) synthesizing the various types of interventions and outcomes used, (2) evaluating the effectiveness of the interventions, and (3) identifying the strengths and limitations of prior studies to make recommendations for evidence-based clinical practice and future research. METHODS: An exhaustive literature search was performed for experimental studies conducted in the past 10 years. FINDINGS: Analysis using 15 identified studies indicated the following strengths: (1) use of random assignment in many of the studies, (2) use of theoretical frameworks to guide the interventions, (3) replication of previous studies, and (4) inclusion of outcome measures of alcohol use, quantity, and frequency. Limitations included: (1) small convenience samples; (2) use of multiple tools to elicit outcomes, making it difficult to compare results across studies; (3) lack of long term follow-up to assess sustainability of the interventions; (4) use of only self-report outcome measures, which rely on subject's recall memory; (5) lack of manipulation checks to assure that subjects actually processed the interventions; and (6) a paucity of stress and coping interventions. CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: Extensive research to address the problem of college alcohol use indicates that while education is an integral part of the approach for this problem, it is ineffective when used alone as an intervention strategy. However, some empirical support exists for the use of brief motivational interventions to reduce alcohol use and harm. A personalized approach addressing expectancies and normative use employing a motivational interviewing style may produce desired outcomes. In addition, theory-based manualized approaches using stress and coping intervention strategies need to be developed and tested. In the design of future studies, careful attention also should be given to methodological issues such as sampling, measurement issues, and inclusion of more long-term follow-up measures. PMID- 17129325 TI - A knowledge transfer strategy for public health decision makers. AB - PURPOSE: The purpose of this study was to discover public health decision makers' preferences for content, format, and channels for receiving research knowledge, so as to begin development of a comprehensive national public health knowledge transfer strategy. A preliminary knowledge transfer strategy developed in part from the views expressed by public health decision makers in an earlier study (Dobbins et al. 2002b) was used as a foundation on which to base discussions. The research team believes strongly that consultation with potential users is crucial to ensure the conduct of relevant and timely research as well as the development of an effective knowledge transfer strategy. METHODS: Nine focus groups of five to seven participants were held in seven Canadian cities. Participants included medical officers of health, public health managers and directors, health promotion mangers, and health policymakers at provincial and federal levels. A semi-structured, open-ended interview guide was used to facilitate the discussion. The focus groups were audiotaped, and results were analyzed independently by two members of the research team who then developed key themes through a consensus process. RESULTS: Generally, participants spoke positively about the knowledge transfer strategy to which they were exposed. In addition, they supported the development of a registry of reviews evaluating the effectiveness of public health interventions rated by methodological quality of the evidence, with a summary statement of the reviews highlighting the results along with specific implications for practice. Participants also indicated they wanted to receive personalized updates of new reviews in their area of interest. Finally, the results highlighted a significant challenge related to knowledge management indicating opportunities for ongoing professional development and training. CONCLUSIONS: These findings were used to create an online registry of reviews evaluating the effectiveness of public health and health promotion interventions. The registry is one component of a comprehensive national public health knowledge transfer strategy. PMID- 17129326 TI - A qualitative study to identify barriers and facilitators to implementation of pilot interventions in the Veterans Health Administration (VHA) Northwest Network. AB - OBJECTIVE: To identify barriers and facilitators to implementation of pilot interventions designed to improve measurement and management of low-density lipoprotein cholesterol (LDL-c) levels in coronary heart disease patients using the evidence/context/facilitation model of implementation of evidence-based practice. DESIGN: Theory-based conceptual content analysis of structured interviews conducted between January and April 2001. SETTING: Six medical centers in the United States Veterans Health Administration Northwest Network. PARTICIPANTS: Fifty-one of 64 individuals (physicians, nurses, pharmacists, dieticians, quality managers, and other clinical and nonclinical staff) who participated in planning and/or implementing pilot interventions. MAIN FINDINGS: Barriers to successful implementation related primarily to the intervention process and secondarily to characteristics of the intervention context. Interview responses indicated that planning, including identification of resources and assessment of potential barriers and facilitators, was a critical and universally underutilized step in the intervention process. CONCLUSIONS: Organized team process, documented plans for intervention activities, and ongoing evaluation are essential for sustaining intervention activities. A top priority for facilitating interventions should be the development of educational materials, such as "how to" guides, that teach intervention teams how to anticipate barriers and make plans to address them, as well as identifying and fostering local experts in planning and implementing interventions. PMID- 17129328 TI - The prevention of pressure ulcers. PMID- 17129329 TI - Overview of the U.S. invitational conference "Advancing Quality Care through Translation Research". PMID- 17129330 TI - Translation research in long-term care: improving pain management in nursing homes. AB - BACKGROUND: Pain prevalence in nursing homes remains high, with multiple resident, staff, and physician barriers presenting serious challenges to its improvement. AIMS: The study aims were to (1) develop and test a multifaceted, culturally competent intervention to improve nursing home pain practices; (2) improve staff, resident, and physician knowledge and attitudes about pain and its management; (3) improve actual pain practices in nursing homes; and (4) improve nursing home policies and procedures related to pain. METHODS: A multifaceted, culturally competent intervention was developed and tested in six Colorado nursing homes, with another six nursing homes serving as control sites. Both educational and behavioral change strategies were employed. FINDINGS: The intervention was successful in improving the percentage of residents reporting constant pain in the treatment homes. Contextual factors (implementation of Medicare's Nursing Home Compare report card) appeared to exert a positive influence on pain documentation. There was no reduction in the percentage of residents reporting pain or reporting moderate/severe pain. DISCUSSION: Multiple challenges to quality improvement exist in nursing homes. Turnover of nursing staff reduced actual exposure to the intervention, and turnover of directors of nursing influenced constancy of message and overall facility stability. Residents often failed to report their pain, and physicians were reluctant to alter their prescribing practices. IMPLICATIONS: Any intervention to improve pain management in nursing homes must target explicitly the residents, nursing home staff, and primary care physicians. Implementation strategies need to accommodate the high turnover rates among staff, as well as the changes among the nursing home leadership. CONCLUSIONS: Pain is a complex problem in the nursing home setting. Multiple factors must be considered in both the design and implementation of interventions to improve pain practices and reduce pain prevalence in nursing homes. PMID- 17129331 TI - Advancing quality of urinary incontinence evaluation and treatment in nursing homes through translational research. AB - AIM: This article presents the development of ongoing research to improve the evaluation and treatment of urinary incontinence (UI) in nursing homes through "A Model for Use of the Urinary Incontinence Guideline in US Nursing Homes" under the initiative Translating Research into Practice (TRIP II). BACKGROUND: The previous research provided the definition of the clinical practices needing improvement, the relative size of the population needing care, and an estimate of the manpower required. The model proposed nurse practitioners (NPs) as consultants to nursing homes to perform the basic incontinence evaluation and treatment, thereby (1) improving incontinence evaluation and treatment, (2) reducing UI, (3) reducing complications, and (4) increasing cost savings attributable to UI. APPROACH: A quasi-experimental research design with pre- and post-comparison was selected utilizing four control and four experimental NP caseloads followed for 16 weeks post-onset (n = 320). FINDINGS: Findings are not yet available from this study, which is ongoing. DISCUSSION: The real-world challenges presented by this translational research and their solutions are discussed. PMID- 17129332 TI - Testing a collaborative research utilization model to translate best practices in pain management. AB - While millions of dollars are being spent developing the seeds of research to improve the care we give to patients, significant barriers to using empirical evidence by health care professionals and policymakers still exist. The gap between what we know from research and what we use in practice is at the heart of the translation research problem. How do we get empirical knowledge uptake and use by clinicians in the fields of practice? In addition, how do we measure if practice changes are occurring? This article describes a program of translation research in which the intervention, called the Collaborative Research Utilization model, was developed and tested in four clinical studies over 18 years. PMID- 17129333 TI - Improving care to patients with ischemic heart disease: experiences in a single network of the Veterans Health Administration. AB - BACKGROUND: Ischemic heart disease (IHD) is the leading cause of death in the United States. Lowering serum cholesterol levels reduces coronary events and mortality; this effect is most evident in patients with preexisting IHD. AIMS: The primary aim of this article is to describe a set of interventions that were piloted in a single, regional Veterans Integrated Service Network (VISN) to promote secondary prevention among patients with IHD and to explore the effect of those interventions on patient outcomes. METHODS: An observational, before-and after study of clinical interventions to improve lipid guideline compliance in VISN 20 (the Veterans Administration Northwest Network) was conducted. A total of 2,467 patients with established coronary artery disease from three medical facilities in VISN 20 were included. Each medical facility chose different interventions to lower low-density lipoprotein cholesterol (LDL-c) levels in their patients. One facility chose a paper point-of-care reminder, a second chose a lipid clinic, and a third chose audit/feedback to clinicians in addition to a patient-education component. Data came from a relational database that mirrors the clinical information system at each site. Outcomes included the proportion of patients who had their LDL-c measured, the proportion of patients who had lipid lowering agents prescribed, and the proportion of patients at LDL-c goal of lower than 100 mg/dL measured before, during, and after the intervention period. RESULTS: Statistically significant improvements were observed within sites after the interventions were implemented. IMPLICATIONS FOR PRACTICE: Interventions that focused on secondary prevention in this high-risk group were moderately successful in changing practice. Tailoring interventions to the needs of a specific site of care is feasible and may add to the likelihood of succeeding. CONCLUSION: Overall, the three facilities improved in lipid measurement and management for patients with coronary artery disease. PMID- 17129334 TI - Outcomes of adoption: measuring evidence uptake by individuals and organizations. AB - BACKGROUND: The translation and diffusion of findings into health care validate the potential of evidence-based innovation to improve clinical practice and affirm the benefits of society's investment in advancing science. AIMS: This article briefly reviews key concepts in the knowledge use process, considers theoretical implications for measuring outcomes and uptake of innovation, discusses issues to consider in planning for measurement of adoption, and provides an example of confronting those challenges from a project now in progress. THEORETICAL PERSPECTIVES AND TRANSLATION OUTCOMES: Four principal conceptual frameworks related to translational science, Lewin, Rogers, Havelock, and Promoting Action on Research Implementation in Health Services (PARIHS), explicate a process that catalyzes new knowledge adoption and use by individuals and systems to solve problems. Each conceptual perspective suggests that translation is not complete until the extent and impact of use is examined and understood. Most perspectives support evaluation of impact of evidence-based practice using process measures that integrate clinician knowledge, actual performance of the practice, and patient/clinician outcomes. Based on the Agency for Healthcare Research and Quality (AHRQ) framework, additional measures might include changes in patterns of care and changes in policies, procedures, or protocols. CASE EXAMPLE: A description of process and outcome measures used in an in-progress quality improvement demonstration project, California Nursing Outcomes Coalition (CalNOC) Partners in Quality Translating Research into Practice to Reduce Patient Falls Project, is presented. IMPLICATION(S) FOR PRACTICE, RESEARCH, AND POLICY: Since the adoption of evidence-based innovation is a process that is often described along a continuum, investigators seeking to measure the impact of an evidence-based innovation must gather evidence that uptake of the innovation has occurred. The theoretical perspective and practical measurement issues of a given project will drive selection of process and outcome measures. CONCLUSIONS: Efforts to change practice in order to enhance evidence based patient care must integrate monitoring and evaluation of specific target outcomes of adoption as the basis for validating the impact of the change. PMID- 17129335 TI - Organizational research with impact: working backwards. AB - BACKGROUND OR RATIONALE: To improve health care, we need to improve the organization of care, along with the payment systems that shape organizational priorities and behavior. The opportunity and challenge for research are to find a way to work with health care leadership so that future management decisions can make use of strong evidence. AIMS AND METHODS: This article uses findings from research on nursing to illustrate the potential for organizational research and management research to improve health care. It then distills recommendations from six focused stakeholder meetings to identify five ways in which we might improve organizational, management, and policy research to maximize its use. RESULTS AND DISCUSSION: Hospital, health plan, and other system leaders have five recommendations for research: (1) Design studies that answer user questions, with a focus on the "why" and "what if" rather than just the "what." (2) Present findings in leaders' time and space, defining evidence as they do and identifying generalizability of findings. (3) Change the incentive system for researchers so that they are rewarded for the activities that maximize impact on decision making. (4) Build user-researcher collaborations and dialogue. (5) Change the way we disseminate evidence, with dissemination through "early adopters," trade association meetings, consultants, etc. IMPLICATIONS FOR RESEARCH, PRACTICE, AND POLICY: System and policy leaders control important levers for improving health care, since they shape organizational structure, processes and culture, payment strategies, program design, and regulation. Just as evidence-based medicine can improve clinical practice, evidence-based management and policymaking can change how these powerful levers are used. But for evidence to inform the decisions of system and policy leaders, we will need to rethink and restructure the research enterprise itself, bringing the potential users of evidence into the production process. PMID- 17129336 TI - State of the science of translational research: from demonstration projects to intervention testing. AB - An overview of the effort of nursing in translation research begins with studies in research utilization. Delineation of issues in the uptake of evidence using guidelines and systematic reviews is discussed next. The federal initiatives in translation research are described with a focus on the Translating Research Into Practice (TRIP) studies. The article closes with future directions. PMID- 17129337 TI - Quantitative analytical methods in translation research. AB - In research involving quality of health care, an intervention is often attempted at one or more institutions. This article presents the advantages and disadvantages of various study designs that might be considered for such institution-based research, looking at the issues of comparison groups, replication, blocking, and contamination. Three examples illustrate the importance of accommodating the lack of independence or clustering that occurs among subjects within an institution, and show how these accommodations for clustering can sometimes be made by applying basic statistical techniques to institution-level outcomes, whereas at other times more sophisticated approaches are necessary. PMID- 17129338 TI - Qualitative perspectives in translational research. AB - The rapid uptake of qualitative approaches in translational research can be best understood in the context of recent innovations in health services research, as well as an overarching concern with improving the quality of health care. Qualitative approaches highlight the human dimension in health care by foregrounding the perceptions, experiences, and behaviors of both consumers and providers of care. As such, these methods are particularly useful for addressing the complex issues related to improving health care quality and implementing system change. This overview traces a brief history of the factors contributing to the recent and rapid growth of qualitative methods in health research in general and translational research in particular; describes the varieties of qualitative approaches employed in this research; and illustrates the utility of these approaches for variable identification, instrument development, description/explanation of patient/provider perceptions and behaviors, individual/organizational change, and theory refinement. PMID- 17129339 TI - Preparing the next generation of scientists in translation research. AB - AIM: Four questions about translation research are considered in this article. (1) Why should there be concern about preparing nurse scientists for translation research? (2) Are current research-intensive doctoral programs in nursing preparing graduates for translation research? (3) Should targeted efforts be made to systematically prepare nurses in translation research? (4) What strategies would be useful? QUESTION 1: It is argued that translation research is needed in order to increase the odds that interventions found to be efficacious can be effectively translated into practices that will produce desired outcomes. Thus, evidence from translation research is a very important component of the science base needed to guide innovation in practice. Another reason of concern in preparing translation researchers is the growing pace of research into the efficacy of interventions, which accelerates the need for translation research. Furthermore, the emergence of clinical doctoral programs in nursing that will be seeking faculty with expertise in translation research supports the need for preparing scientists in translation research. QUESTION 2: To determine whether translation research was visible in current research-intensive doctoral programs, a Web-based review of these programs offered by colleges and universities in the database of the American Association of Colleges of Nursing (AACN) was conducted in the fall of 2003. The search revealed that in the 79 programs surveyed, no school identified an area of emphasis in terms that suggested a component of translation science such as research dissemination or utilization, knowledge transfer, diffusion of innovation, or technology transfer. Less than a dozen courses could be located that focused on topics related to translation research or translation science. QUESTIONS 3 AND 4: The case is made that more attention should be given to preparing translation scientists in research-intensive doctoral programs in nursing, and several strategies were suggested for moving toward that goal. PMID- 17129340 TI - Translation research: where do we go from here? AB - AIM: Where do we go from here? This article draws on other articles in this supplement, the dialogue from the meeting that generated it, and other sources to identify steps to advance translation research, and in particular to achieve broader translation and use of evidence. IMPLICATIONS: To move from the growing accumulation of individual successes in translation to broader scale translation and implementation of evidence, those who use research and those who do research will need to do four things: (1) increase synergy and synchronization across studies, through cross-disease studies, adoption of a common and precise language to describe interventions, addressing issues of customization of interventions, and finding commonality in quality and outcomes measures; (2) take account of organizational factors that can shape the impact of interventions, and also move to the implementation of organizational evidence; (3) address environmental factors such as reimbursement and market competition; and (4) take findings to a larger scale through national demonstrations, efforts of national agents of change, challenge or partnership grants, or use of provider-based networks. PMID- 17129342 TI - Tissue restoration: approaches and prospects. AB - This article summarizes the results of three basic research approaches directed toward achieving the restoration of injured or diseased human tissues. The three approaches are (1) to understand the differences in the molecular characteristics of cells and their environments in tissues which exhibit regenerative capacity at one stage of the life cycle but not at another; (2) to understand the molecular mechanisms whereby tissues regenerate by means of reserve progenitor cells or progenitor cells formed by dedifferentiation; and (3) to design artificial tissues for implantation into the body. These strategies should allow us to locate the key switchpoints which determine regeneration versus repair and how to reconfigure those switchpoints into a regenerative circuit where necessary or to build tissues in vitro that can serve as in vivo substitutes. For these strategies to be successful, we must understand the role of the immune system in repair and regeneration and the developmental roles of regulatory molecules, such as growth factors, trophic factors, extracellular matrix components, and the protein products of patterning genes, as well as the intracellular signaling systems activated by these molecules. The examples used to illustrate these themes are repair versus regeneration in wounded mammalian skin, the regeneration of injured mammalian bone and muscle by reserve progenitor cells, and the regeneration of the urodele limb, neural retina, and lens by progenitor cells produced by dedifferentiation. In addition, several approaches used in the construction of bioartificial tissues are discussed. PMID- 17129343 TI - Inhibition of tissue transglutaminase and epsilon (gamma-glutamyl) lysine cross linking in human hypertrophic scar. AB - Immunohistochemical staining with monoclonal antibody to tissue transglutaminase was used to study cryostat sections of human skin wounds. The enzyme was found in acute wounds and chronic hypertrophic scars but not in normal mature scars. Because tissue transglutaminase is responsible for the formation of isopeptide cross-links, a two-stage high-performance liquid chromatographic analysis was used to quantitate the epsilon (gamma-glutamyl) lysine cross-link produced by various types of wound tissues. Eighteen patients with hypertrophic scars between 6 months' and 10 years' duration after injury underwent a double-blind trial with putrescine 50 mmol/L in a eutectic vehicle for 2 months under nonocclusive dressings (Biofill). For the control portion of the same or different scar, sham vehicle and non-occlusive dressing were simultaneously applied. Both scars were harvested at biopsy or elective revision surgery 2 months later. After homogenization and exhaustive proteolysis, digests were studied with the use of high-performance liquid chromatography analysis. The results of treatment were a significant decrease in the levels of isopeptide cross-link formation from 0.018 +/- .006 nmol/micromol amino acids in untreated scars to 0.008 +/- .001 nmol/micromol amino acid in the treated group (p < 0.05). The isopeptide cross link content in treated scars was nearly as low as that in normal mature scars (0.003 +/- 0.001 nmol/micromol amino acid). These results show that cross-link formation by tissue transglutaminase activity is inhibited during treatment of hypertrophic scar by putrescine. These results support the possible therapeutic use of topical putrescine in the treatment of hypertrophic scar formation. PMID- 17129344 TI - Effect of electrical stimulation waveform on healing of ulcers in human beings with spinal cord injury. AB - Various electrical stimulation waveforms have been used to enhance wound healing, with little consideration for potential differences in their physiologic effect. The present study evaluated the effect of stimulation waveform and electrode placement on wound healing. Eighty patients with spinal cord injury and one or more pressure ulcers were treated. A total of 185 ulcers received 45 minutes of stimulation daily. Each ulcer was subjected to one of four treatment protocols: asymmetric biphasic waveform, symmetric biphasic waveform, microcurrent stim ulation, or a sham control protocol. Electrodes were placed outside the wounds, over intact skin and surrounding the area of the ulcer. Data were categorized by ulcers which healed during the protocol and those which did not. Analysis of the "good response" ulcers (n = 104) showed significantly better healing rates for those receiving stimulation with the asymmetric biphasic waveform, compared with the control and microcurrent groups. Mean healing rates from the present study were similar to previously reported measures. The waveforms studied possessed minimal polar capabilities, and the electrodes were placed outside the wound. These data show that electrical stimulation clearly enhanced healing of pressure ulcers in a significant number of individuals with spinal cord injury; the physiologic implications of these findings relative to the mechanism(s) by which electrical stimulation enhances wound healing are discussed. However, extrapolation of these results to patients with other types of wounds must await further study. PMID- 17129345 TI - Wound contraction and scar synthesis during development of the amphibian Rana catesbeiana. AB - Certain characteristics of skin wound healing in the North American bullfrog, Rana catesbeiana, were studied grossly and histologically at various stages of development. The contraction kinetics of excisional skin wounds were monitored for 50 days in animals from four larval developmental stages (tadpoles) and from adults (frogs). The percentage of original wound area which remained after contraction had stopped in the four larval groups decreasing steadily from 59.2%+/- 6.8% for the least developed larvae to 9.9%+/- 2.3% for the most developed ones. Wounds in adults contracted at a much lower rate than for larvae and the percentage of original wound area remaining after 50 days was less than 10%. The morphologic features characterizing the intact dermis and epidermis outside the wound bed in the larvae were also observed in qualitatively similar detail inside the wound bed. In the adults, the subepidermal connective tissue inside the wound bed was distinctly different from the physiologic dermis outside and was classified as amphibian scar. We conclude that, during larval development, wound contraction increasingly displaced skin regeneration as a mechanism for wound closure. Adults closed skin wounds by a combination of contraction and synthesis of scar tissue. It is hypothesized that the mechanism for synthesis of scar in this amphibian is established during metamorphosis from larva to adult. PMID- 17129346 TI - Growth hormone increases the biomechanical strength and collagen deposition rate during the early phase of skin wound healing. AB - The effects of biosynthetic human growth hormone on the biomechanical properties and collagen deposition rate of wound healing were investigated in rat skin incisional wounds after 4 days of healing. Biosynthetic human growth hormone induced a pronounced, < or =94%, and dose-dependent increase in the mechanical strength of wounds in the dose range of 0.125 to 2 mg/kg/day. A new method for in vivo studies of the collagen deposition rate in granulation tissue of the wound cleft was applied. The production of (3)H-hydroxy-l-proline was measured by injecting (3)H-proline intravenously into the rats with a large flooding dose of unlabeled proline which reduces reutilization of (3)H-proline and reduces the influence of de novo synthesis of proline. Extractable collagens, which are not bound in the wound tissue and therefore do not contribute mechanical strength to it, were removed from the samples. Labeled and unlabeled proline were determined simultaneously by reverse-phase high-performance liquid chromatography with ultraviolet detection and flow scintillation counting of these amino acids. At day 4 the collagen deposition rate in the incisional wound zone, 0.4 mm wide containing the wound cleft, was 1.8% per hour of the collagen present in the wound zone. The collagen deposition rate was increased by 149% by biosynthetic human growth hormone 2 mg/kg/day compared with the control group. This result indicates that the increased biomechanical strength of the skin incisional wounds of the groups treated with biosynthetic human growth hormone was produced by an increased deposition of collagen in the wound cleft. PMID- 17129347 TI - Analysis of the effects of chitosan on inflammation, angiogenesis, fibroplasia, and collagen deposition in polyvinyl alcohol sponge implants in rat wounds. AB - Chitosan is a large molecular weight, positively charged polysaccharide extracted and purified from the chitin of crab shells. This compound has been shown to have hemostatic activity and has been suggested for use as a topical agent in tissue repair. The objective of this study was to analyze the effect of chitosan on the wound healing response to a standardized injury in the rat. The polyvinyl alcohol sponge implant model was used as a means to deliver either chitosan or its vehicle to a standardized subcutaneous wound on the backs of Sprague-Dawley rats. On days 8 and 14, the chitosan-treated implants contained primarily polymorphonuclear leukocytes compared with the vehicle controls which contained mainly macrophages, fibroblasts, collagen, and new blood vessels. High performance liquid chromatography analysis for hydroxy-L-proline deposited in the sponge implants showed significantly lower amounts on both days 8 and 14 in the chitosan treatment group. These histologic and biochemical studies suggest that chitosan modulates wound healing by first reducing the influx of activated tissue macrophages, which in turn reduces the subsequent events of angiogenesis, fibroplasia, and connective tissue deposition. PMID- 17129348 TI - Prevention of postoperative adhesions with the chitin derivative N-O carboxymethylchitosan. AB - The ideal barrier agent for the prevention of surgical adhesions has remained elusive. We have examined the ability of a new hydrogel N-O carboxymethylchitosan, a derivative of chitin with properties similar to the extracellular matrix, to prevent adhesions when applied topically to traumatized mesothelial surfaces. In two rodent adhesion models (pericardial and peritoneal), the application of N-O-carboxymethylchitosan significantly prevented or minimized the formation of scar and fibrosis. According to a scoring system from 0 to 3 (0 = no adhesions and 3 = severe dense adhesions), control groups in each model consistently produced severe dense adhesions (2.9 +/- 0.2, 2.7 +/- 0.3). All treated groups consistently scored less than 1.0, indicating minimal or no fibrosis. The differences between the control and treated groups were statistically significant (p < 0.05). Thus, the application of N-O carboxymethylchitosan to traumatized mesothelial surfaces may have significant potential in the prevention of postoperative adhesion formation. PMID- 17129349 TI - Effect of growth factors on the characteristics of cells associated with equine wound healing and sarcoid formation. AB - Wound healing in equidae is delayed and more complicated than in other species. These complications arise from a condition known as exuberant granulation tissue formation. The lower limb of the horse is frequently slower to heal than other parts of the body and has a particular tendency to produce excess (exuberant) granulation tissue. Sarcoids are tumor-like lesions of the skin which often appear at the site of wounds. This study compared the growth characteristics of the sarcoid and granulation tissue-derived cells with normal dermal fibroblasts grown from primary cell cultures. All three cell types had distinct morphologic differences. Growth rate studies showed that the sarcoid and granulation tissue derived cells grew at a slower rate than the normal cells. The addition of the growth factors epidermal growth factor, acidic fibroblast growth factor, and basic fibroblast growth factor selectively stimulated the replication of normal and sarcoid-derived cells but inhibited the growth of granulation tissue-derived cells. In contrast, transforming growth factor-beta was not preferentially inhibitory for the granulation tissue-derived cells. The addition of growth factors to the medium also produced distinct alterations in the morphology of the cells. PMID- 17129350 TI - Fetal wound healing: an overview. AB - The ability of fetal tissues to heal without scarring has prompted extensive research into the biochemical and molecular differences between fetal and postnatal wound healing. A thorough understanding of the basic mechanisms of fetal wound repair may to lead to approaches to correct or prevent the clinical problems encountered in abnormal adult wound healing and fetal surgery. This article contrasts the normal healing response in adults with fetal repair in animal models, highlighting investigations of extracellular matrix expression, cytokine profiles, and cellular dynamics. PMID- 17129351 TI - Extracellular matrix protein turnover during salamander limb regeneration. AB - After amputation of a salamander limb, the extracellular matrix undergoes remodeling. The extracellular matrix that maintains the differentiated state of limb tissues is broken down and replaced by an extracellular matrix essential for dedifferentiation and blastema formation. We used monoclonal antibodies in immunohistochemistry methods and riboprobes in in situ hybridization to evaluate the upregulation of tenascin, type XII collagen, fibronectin, and the MT5 antigen. The Stump 1 antigen, an extracellular matrix protein that is abundant in the normal limb, is downregulated during regeneration and reappears late in regeneration as differentiation occurs. In the embryo, the Stump 1 antigen is also absent from the early limb bud and first appears during differentiation stages. Tenascin and fibronectin are also upregulated in the limb bud of the embryo, and these two extracellular matrix proteins appear to function during limb regeneration in adults and limb development in embryos. However, type XII collagen and the MT5 antigen are not found in the limb bud, indicating that type XII collagen and the MT5 antigen have roles in the regenerating limb but not in the embryo limb bud. PMID- 17129352 TI - Expression of bone morphogenetic protein-4 mRNA during the process of bone formation in embryogenesis and fracture repair. AB - Bone morphogenetic protein-4 is a potent inducer of ectopic bone and cartilage formation in vivo. Expression of the bone morphogenetic protein-4 gene has been detected in bone cells during fracture repair but not in normal adult bone cells. To examine whether the gene is expressed by bone cells during embryonic development, in situ hybridization and reverse transcription polymerase chain reaction methods were used to detect murine bone morphogenetic protein-4 specific complementary DNA in murine developmental bone tissues. Bone morphogenetic protein-4 messenger RNA was detected in the cells of various developing bone tissues, but it was not detected in these tissues after birth. Combined with previous reports, our findings indicate that the bone morphogenetic protein-4 gene is expressed during embryogenesis and bone repair and suggest that its product may be a potent bone-forming factor in bone development and fracture repair. PMID- 17129353 TI - Progression of cellular repopulation and collagen synthesis in fresh-frozen allograft tendons. AB - The progression of cellular repopulation and collagen synthesis in fresh-frozen rat patellar tendon allografts was investigated by means of indirect immunofluorescence histologic analysis and electron microscopic techniques for 8 weeks after transplantation. In each of 10 procedures, the medial half of the patellar tendon with a tibial bone block was harvested from a Wistar rat and transplanted into a corresponding defect in the medial patellar tendon of a Lewis rat. Actin filaments in the repopulating cells and newly synthesized collagen fibrils in the graft were identified with rhodamine-phalloidin stain and with a polyclonal antibody against type III collagen aminopropeptide. On the first day after transplantation, no specific fluorescence was detected in the graft. One week later, specific labeling for fibrillar-actin (F-actin) and type III collagen aminopropeptide was detected in an area extending from the adjacent granulation tissue into the proximal end of the graft. F-actin and type III collagen aminopropeptide were aligned along the longitudinal axis of the graft and extended from the proximal suture site toward the distal portion. Two weeks after transplantation, fibrillar labeling for F-actin and type III collagen amino propeptide showed that remodeling had extended to the midportion of the graft. Labeling throughout the entire graft was detected 4 weeks after transplantation. During the entire remodeling process, the repopulated fibroblasts consistently retained their elongated shape and their alignment with the longitudinal axis of the graft. The cells developed well-organized actin bundles at their peripheries, which identified them as having a myofibroblast phenotype. Immunofluorescence detection for type III collagen aminopropeptide also showed consistent alignment parallel to the longitudinal axis of the graft and a fibrillar arrangement. Electron microscopy revealed thinner collagen fibrils in the vicinity of the fibroblasts, which were aligned in the direction of the actin bundles. These results indicate that, during the early remodeling phase, collagen synthesis and deposition in the graft proceeds while the original alignment of the graft matrix is preserved. The close association between the alignment of actin bundles in repopulated "myofibroblastic" cells and that of newly synthesized collagen fibrils along the lines of the graft matrix may represent evidence of force transmission between the actin cytoskeleton and the linking extracellular matrix in vivo. PMID- 17129354 TI - Mesenchymal-epithelial interactions regulate gene expression of type VII collagen and kalinin in keratinocytes and dermal-epidermal junction formation in a skin equivalent model. AB - Anchoring fibrils constituted primarily of type VII collagen and anchoring filaments composed of kalinin are essential structural elements of the dermal epidermal junction and critical for its stability. The role of fibroblasts in the production of these structural elements and the formation of the dermal-epidermal junction was studied by using a living skin equivalent model. This model had been modified such that keratinocytes and fibroblasts were allowed direct contact. After 2 weeks, immunohistochemical studies showed the linear deposition of type VII collagen and kalinin, as well as type IV collagen and alpha6 integrin at the dermal-epidermal junction. By electron microscopy, anchoring fibrils, a continuous lamina densa, and numerous hemidesmosomes were noted. Reverse transcriptase-polymerase chain reaction analysis showed an increased expression of both type VII collagen and kalinin genes in keratinocytes when they were in direct contact with fibroblasts. These results suggest that fibroblasts synthesize an extracellular matrix which favors keratinocyte adhesion and the formation of a dermal-epidermal junction by increasing the production and the further arrangement of dermal-epidermal junction components. PMID- 17129355 TI - Characteristics of growth, morphology, contractility, and protein expression of fibroblasts derived from keloid. AB - Phenotypic alterations of keloid-derived fibroblasts were characterized by comparison with the phenotypes of normal fibroblasts from the same patient. Explant cultures of keloids showed unique features. Keloid explants contracted considerably and reduced their size during culture, whereas the size of normal skin explants remained unchanged. Enlarged cells were found among fibroblasts which had grown out of all the explants and were morphologically distinct from fibroblasts; however, keloid explants produced many more of them than did the normal tissues. The growth rate of fibroblast colonies formed from normal explants was five times higher than keloid explants. Keloid fibroblasts which had been serially cultivated contracted lattices of collagen gels at a rate similar to normal fibroblasts. Proteins extracted from serially cultivated fibroblasts were mapped on polyacrylamide two-dimensional electrophoretic gels. No significant qualitative alterations in protein expression in keloid cells were found as compared with normal fibroblasts. However, some quantitative changes were found between the two. A computer-assisted image analyzer detected 151 polypeptide spots--50 spots (33%) of which increased their amounts in keloid cells, whereas 34 spots (22.5%) decreased in comparison with normal fibroblasts. Sixteen major polypeptides were identified as known proteins with the aid of time of-flight mass spectrometry. The level of expression of these identified proteins was similar between normal and keloid cells, except stathmin whose expression was suppressed in keloid fibroblasts. PMID- 17129356 TI - Expression of proliferating cell nuclear antigen in cultured middle ear epithelial cells of the guinea pig. AB - Primary cultures of middle ear epithelium from the guinea pig were successfully established on type I collagen coated dishes. To characterize cellular outgrowth, antibodies to the proliferating cell nuclear antigen were used as a marker for spreading cells in the S phase of the cell cycle. A number of migrating epithelial cells positively stained for proliferating cell nuclear antigen after 7 and 14 days in culture. Confocal laser scanning microscopy was used to evaluate the localization pattern of this antigen, and the fluorescence intensity was quantified in different areas of the migrating epithelial sheet after various times in culture. Two distinct areas proved to be major sites of proliferating cell nuclear antigen expression. One was at the edge of the tissue explants from which multilayered epithelial cells had begun to migrate. The other was along the margin of the outgrowth, where the cells often had elongated shapes and were aligned in rows. The cells in both areas were identified as nonciliated cells; ciliated cells in the outgrowth showed little staining. We hypothesized that the outgrowth cells in this experiment might be identical to the migrating cells usually observed in renewing epithelia after injury. This model may provide a simple and reproducible method of evaluating the regenerative ability of the middle ear epithelium. PMID- 17129358 TI - Up-regulation of a BCL9-related beta-catenin-binding protein, B9L, in different stages of sporadic colorectal adenoma. AB - Aberrant activation of Wnt signaling is a critical event in the development of human colorectal tumors. The aim of this study was to elucidate the role of B9L and its association with beta-catenin in each stage of the adenoma-carcinoma sequence of human colorectal tumorigenesis. We investigated the expression levels of B9L in sporadic colorectal adenomas and carcinomas, categorized according to the Vienna classification, using real-time quantitative polymerase chain reaction (RTQ-PCR) and immunohistochemical analysis. B9L was expressed in the nuclei of non-neoplastic colonic mucosa cells and was overexpressed in the nuclei of neoplasias and invasive carcinoma cells. Immunoreactivity to B9L protein was correlated with the progressive grades of colorectal neoplasias, as was the expression of B9L mRNA. A high level of immunoreactivity to nuclear B9L was present in 27% of low-grade neoplasias and in more than 50% of high-grade neoplasias and invasive carcinomas, whereas a high level of nuclear B9L immunoreactivity was not observed in any of the non-neoplastic mucosa samples. The expression of B9L was dramatically increased in low-grade neoplasias compared with that in non-neoplastic mucosa. B9L may play an important role in tumorigenesis induced by aberrant activation of Wnt signaling and may act as a key protein in the progressive dysplasia of adenoma. PMID- 17129359 TI - Zerumbone, a bioactive sesquiterpene, induces G2/M cell cycle arrest and apoptosis in leukemia cells via a Fas- and mitochondria-mediated pathway. AB - We demonstrated here for the first time that zerumbone (ZER), a natural cyclic sesquiterpene, significantly suppressed the proliferation of promyelocytic leukemia NB4 cells among several leukemia cell lines, but not human umbilical vein endothelial cells (HUVECs), by inducing G2/M cell cycle arrest followed by apoptosis with 10 microM of IC50. Treatment of NB4 cells with growth-suppressive concentrations of ZER resulted in G2/M cell cycle arrest that was associated with a decline of Cyclin B1 protein, but with the phosphorylation of ATM/ Chk1/Chk2. In addition, ZER induced the phosphorylation of Cdc25C at the Thr48 residue and Cdc2 at the Thr14/Tyr15 residues. Furthermore, ZER-induced apoptosis in NB4 cells was initiated by the expression of Fas (CD95)/Fas Ligand (CD95L), concomitant with the activation of caspase-8. ZER was also found to induce the cleavage of Bid, a mediator that is known to connect the Fas/CD95 cell death receptor to the mitochondrial apoptosis pathway. ZER also induced the cleavage of Bax and Mcl-1 proteins, but not Bcl-2 or Bcl-XL. ZER-induced apoptosis took place in association with a loss of the mitochondrial transmembrane potential as well as the activation of caspase-3 and -9, resulting in the degradation of the proteolytic poly (ADP-ribose) polymerase (PARP). ZER also triggered a release of cytochrome c into the cytoplasm. Both antagonistic anti-Fas antibody ZB4 and pan caspase inhibitor Z-VAD inhibited ZER-induced apoptosis in NB4 cells. Taken together, ZER is an inducer of apoptosis in leukemic cells that specifically triggers the Fas/CD95- and mitochondria-mediated apoptotic signaling pathway. PMID- 17129360 TI - Carcinogenesis and transcriptional regulation through Maf recognition elements. AB - Many studies on carcinogenesis carried out early in the last century are united on the consensus that cancer is a genetic disease. Cancer cells typically display gene dysfunction and endogenous or exogenous insults resulting in gene dysfunction are often carcinogenic. Recent advances in stem cell biology added the new concept that cancer originates from a single cancer-initiating cell. To understand the molecular basis of carcinogenesis from the beginning to the full acquirement of malignancy, factors concerned with carcinogenesis were categorized into three groups: those guarding and stabilizing genomes, those regulating cell proliferation, and those conferring resistance to various micro-environmental stresses. One example of particular interest is the Keap1-Nrf2 system since, according to recent studies, it has turned out to be ambivalent. Nrf2 heterodimerizes with small Maf protein to strongly activate transcription through the Maf recognition element (MARE) and Keap1 is an inhibitory regulator of Nrf2. The genes regulated by Nrf2 are very important for cellular protection of the genome from xenobiotic and oxidative stresses and, consequently, for preventing carcinogenesis. This implies that enhancing Nrf2 activity is a promising method for thwarting cancer. On the contrary, the constitutive activation of Nrf2 due to mutations in the keap1 gene is characteristically observed in lung cancer cells, suggesting that induced expression of Nrf2 target genes favors the prevalence of cancer cells. PMID- 17129361 TI - Ki26894, a novel transforming growth factor-beta type I receptor kinase inhibitor, inhibits in vitro invasion and in vivo bone metastasis of a human breast cancer cell line. AB - Transforming growth factor (TGF)-beta signaling has been shown to promote tumor growth and metastasis in advanced cancer. Use of inhibitors of TGF-beta signaling may thus be a novel strategy for treatment of patients with such cancers. In this study, we investigated the effects of a novel TGF-beta type I receptor (TbetaR-I) kinase inhibitor, Ki26894, on bone metastasis of a highly bone-metastatic variant of human breast cancer MDA-MB-231 cells, termed MDA-MB-231-5a-D (MDA-231-D). Ki26894 blocked TGF-beta signaling in MDA-231-D cells, as detected by suppression of phosphorylation of Smad2 and inhibition of TGF-beta-responsive reporter activity. Moreover, Ki26894 decreased the motility and the invasion of MDA-231-D cells induced by TGF-beta in vitro. Ki26894 also suppressed transcription of plasminogen activator inhibitor-1 (PAI-1), parathyroid hormone-related protein (PTHrP), and interleukin-11 (IL-11) mRNA of MDA-231-D cells, which were stimulated by TGF-beta. X-ray radiography revealed that systemic Ki26894 treatment initiated 1 day before the inoculation of MDA-231-D cells into the left ventricle of BALB/cnu/nu female mice resulted in decreased bone metastasis of breast cancer cells. Moreover, Ki26894 prolonged the survival of mice inoculated with MDA-231-D cells compared to vehicle-treated mice. These findings suggest that TbetaR-I kinase inhibitors such as Ki26894 may be useful for blocking the progression of advanced cancers. PMID- 17129362 TI - Can you justify not using ultrasound guidance for central venous access? AB - Karakitsos and coworkers, in this journal, reported further compelling evidence on the value of ultrasound in guiding internal jugular vein catheterization. In a large, prospective, randomized study of 900 patients, comparisons were made between patients in whom the procedure was performed using landmark-based techniques and those assigned to ultrasound guidance. The key benefits from use of ultrasound included reduction in needle puncture time, increased overall success rate (100% versus 94%), reduction in carotid puncture (1% versus 11%), reduction in carotid haematoma (0.4% versus 8.4%), reduction in haemothorax (0% versus 1.7%), decreased pneumothorax (0% versus 2.4%) and reduction in catheter related infection (10% versus 16%). The implications of these findings are discussed, and a compelling case for routine use of ultrasound to guide central venous access is made. PMID- 17129363 TI - 'Chumanzee' evolution: the urge to diverge and merge. AB - A recent analysis of the human and chimpanzee genomes compared with portions of other primate genomes suggests that the divergence of the human and chimpanzee lineages beginning around 6 million years ago was not a simple clean split. PMID- 17129364 TI - Vasopressin in vasodilatory shock: ensure organ blood flow, but take care of the heart! AB - Supplementary arginine vasopressin infusion in advanced vasodilatory shock may be accompanied by a decrease in cardiac index and systemic oxygen transport capacity in approximately 40% of patients. While a reduction of cardiac output most frequently occurs in patients with hyperdynamic circulation, it is less often observed in patients with low cardiac index. Infusion of inotropes, such as dobutamine, may be an effective strategy to restore systemic blood flow. However, when administering inotropic drugs, systemic blood flow should be increased to adequately meet systemic demands (assessed by central or mixed venous oxygen saturation) without putting an excessive beta-adrenergic stress on the heart. Overcorrection of cardiac index to hyperdynamic values with inotropes places myocardial oxygen supply at significant risk. PMID- 17129365 TI - Heterotopic ossification--a long-term consequence of prolonged immobility. AB - Heterotopic ossification is a condition affecting an appreciable minority of critical care patients; it can have long-lasting effects on recovery and return to functional status. Ectopic bone forms in soft tissues near the large joints, causing pain, swelling, limitation of movement and ultimate disability. X-ray changes may be delayed for several weeks after the diagnosis is clinically suspected. Magnetic resonance imaging may be more sensitive for detecting early changes, yielding positive results several weeks before X-rays. However it is not clear that diagnosing the process early will influence long-term patient outcome, because no effective treatments are available. PMID- 17129366 TI - Novel mutation of the PRNP gene of a clinical CJD case. AB - BACKGROUND: Transmissible spongiform encephalopathies (TSEs), a group of neurodegenerative diseases, are thought to be caused by an abnormal isoform of a naturally occurring protein known as cellular prion protein, PrPC. The abnormal form of prion protein, PrPSc accumulates in the brain of affected individuals. Both isoforms are encoded by the same prion protein gene (PRNP), and the structural changes occur post-translationally. Certain mutations in the PRNP gene result in genetic TSEs or increased susceptibility to TSEs. CASE PRESENTATION: A 70 year old woman was admitted to the hospital with severe confusion and inability to walk. Relatives recognized memory loss, gait and behavioral disturbances over a six month period prior to hospitalization. Neurological examination revealed Creutzfeldt-Jakob disease (CJD) related symptoms such as incontinence, Babinski sign and myoclonus. EEG showed periodic sharp waves typical of sporadic CJD and cerebrospinal fluid analysis (CSF) was positive for the presence of the 14-3-3-protein. As the disease progressed the patient developed akinetic mutism and died in the tenth month after onset of the disease symptoms. Unfortunately, no autopsy material was available. PRNP sequencing showed the occurrence of a point mutation on one allele at codon 193, which is altered from ACC, coding for a threonine, to ATC, encoding an isoleucine (T193I). CONCLUSION: Here we report a novel mutation of the PRNP gene found in an elderly female patient resulting in heterozygosity for isoleucine and threonine at codon 193, in which normally homozygosity for threonine is expected (T193). The patient presented typical clinical symptoms of CJD. EEG findings and the presence of the 14-3-3 protein in the CSF, contributed to CJD diagnosis, allowing the classification of this case as a probable CJD according to the World Health Organization (WHO) accepted criteria. PMID- 17129367 TI - SigFlux: a novel network feature to evaluate the importance of proteins in signal transduction networks. AB - BACKGROUND: Measuring each protein's importance in signaling networks helps to identify the crucial proteins in a cellular process, find the fragile portion of the biology system and further assist for disease therapy. However, there are relatively few methods to evaluate the importance of proteins in signaling networks. RESULTS: We developed a novel network feature to evaluate the importance of proteins in signal transduction networks, that we call SigFlux, based on the concept of minimal path sets (MPSs). An MPS is a minimal set of nodes that can perform the signal propagation from ligands to target genes or feedback loops. We define SigFlux as the number of MPSs in which each protein is involved. We applied this network feature to the large signal transduction network in the hippocampal CA1 neuron of mice. Significant correlations were simultaneously observed between SigFlux and both the essentiality and evolutionary rate of genes. Compared with another commonly used network feature, connectivity, SigFlux has similar or better ability as connectivity to reflect a protein's essentiality. Further classification according to protein function demonstrates that high SigFlux, low connectivity proteins are abundant in receptors and transcriptional factors, indicating that SigFlux candescribe the importance of proteins within the context of the entire network. CONCLUSION: SigFlux is a useful network feature in signal transduction networks that allows the prediction of the essentiality and conservation of proteins. With this novel network feature, proteins that participate in more pathways or feedback loops within a signaling network are proved far more likely to be essential and conserved during evolution than their counterparts. PMID- 17129368 TI - Methods for identifying surgical wound infection after discharge from hospital: a systematic review. AB - BACKGROUND: Wound infections are a common complication of surgery that add significantly to the morbidity of patients and costs of treatment. The global trend towards reducing length of hospital stay post-surgery and the increase in day case surgery means that surgical site infections (SSI) will increasingly occur after hospital discharge. Surveillance of SSIs is important because rates of SSI are viewed as a measure of hospital performance, however accurate detection of SSIs post-hospital discharge is not straightforward. METHODS: We conducted a systematic review of methods of post discharge surveillance for surgical wound infection and undertook a national audit of methods of post discharge surveillance for surgical site infection currently used within United Kingdom NHS Trusts. RESULTS: Seven reports of six comparative studies which examined the validity of post-discharge surveillance methods were located; these involved different comparisons and some had methodological limitations, making it difficult to identify an optimal method. Several studies evaluated automated screening of electronic records and found this to be a useful strategy for the identification of SSIs that occurred post discharge. The audit identified a wide range of relevant post-discharge surveillance programmes in England, Scotland and Wales and Northern Ireland; however, these programmes used varying approaches for which there is little supporting evidence of validity and/or reliability. CONCLUSION: In order to establish robust methods of surveillance for those surgical site infections that occur post discharge, there is a need to develop a method of case ascertainment that is valid and reliable post discharge. Existing research has not identified a valid and reliable method. A standardised definition of wound infection (e.g. that of the Centres for Disease Control) should be used as a basis for developing a feasible, valid and reliable approach to defining post discharge SSI. At a local level, the method used to ascertain post discharge SSI will depend upon the purpose of the surveillance, the nature of available routine data and the resources available. PMID- 17129369 TI - Surgical wound infection: epidemiology, pathogenesis, diagnosis and management. AB - Surgical wound infection remains a significant problem following an operation, although surveillance for such infections remains a challenge exacerbated by early discharge and outpatient surgery. The risk of such infections is determined by technical problems with the operation, particularly bleeding, the amount of devitalized tissue created, and the need for drains within the wound, as well as such metabolic factors as obesity and diabetes. Perioperative antibiotic prophylaxis can decrease the incidence of such infections further, but a technically perfect operation is even more important. PMID- 17129370 TI - Altered Ca2+ homeostasis in polymorphonuclear leukocytes from chronic myeloid leukaemia patients. AB - BACKGROUND: In polymorphonuclear leukocytes (PMNL), mobilization of calcium ions is one of the early events triggered by binding of chemoattractant to its receptors. Besides chemotaxis, a variety of other functional responses are dependent on calcium ion mobilization. PMNL from chronic myeloid leukaemia (CML) patients that were morphologically indistinguishable from normal PMNL were found to be defective in various functions stimulated by a chemoattractant - fMLP. To study the mechanism underlying defective functions in CML PMNL, we studied calcium mobilization in CML PMNL in response to two different classical chemoattractants, fMLP and C5a. RESULTS: Release of calcium estimated by flow cytometry and spectrofluorimetry using fluo-3 as an indicator showed that the [Ca2+]i levels were lower in CML PMNL as compared to those in normal PMNL. But, both normal and CML PMNL showed maximum [Ca2+]i in response to fMLP and C5a at 10 sec and 30 sec, respectively. Spectrofluorimetric analysis of the total calcium release in chemoattractant treated PMNL indicated more and faster efflux of [Ca2+]i in CML PMNL as compared to normal PMNL. CONCLUSION: Fine-tuning of Ca2+ homeostasis was altered in CML PMNL. The altered Ca2+ homeostasis may contribute to the defective functions of CML PMNL. PMID- 17129371 TI - Refining motifs by improving information content scores using neighborhood profile search. AB - The main goal of the motif finding problem is to detect novel, over-represented unknown signals in a set of sequences (e.g. transcription factor binding sites in a genome). The most widely used algorithms for finding motifs obtain a generative probabilistic representation of these over-represented signals and try to discover profiles that maximize the information content score. Although these profiles form a very powerful representation of the signals, the major difficulty arises from the fact that the best motif corresponds to the global maximum of a non-convex continuous function. Popular algorithms like Expectation Maximization (EM) and Gibbs sampling tend to be very sensitive to the initial guesses and are known to converge to the nearest local maximum very quickly. In order to improve the quality of the results, EM is used with multiple random starts or any other powerful stochastic global methods that might yield promising initial guesses (like projection algorithms). Global methods do not necessarily give initial guesses in the convergence region of the best local maximum but rather suggest that a promising solution is in the neighborhood region. In this paper, we introduce a novel optimization framework that searches the neighborhood regions of the initial alignment in a systematic manner to explore the multiple local optimal solutions. This effective search is achieved by transforming the original optimization problem into its corresponding dynamical system and estimating the practical stability boundary of the local maximum. Our results show that the popularly used EM algorithm often converges to sub-optimal solutions which can be significantly improved by the proposed neighborhood profile search. Based on experiments using both synthetic and real datasets, our method demonstrates significant improvements in the information content scores of the probabilistic models. The proposed method also gives the flexibility in using different local solvers and global methods depending on their suitability for some specific datasets. PMID- 17129372 TI - Cross-priming of cyclin B1, MUC-1 and survivin-specific CD8+ T cells by dendritic cells loaded with killed allogeneic breast cancer cells. AB - INTRODUCTION: The ability of dendritic cells (DCs) to take up whole tumor cells and process their antigens for presentation to T cells ('cross-priming') is an important mechanism for induction of tumor specific immunity. METHODS: In vitro generated DCs were loaded with killed allogeneic breast cancer cells and offered to autologous naive CD8+ T cells in 2-week and/or 3-week cultures. CD8+ T cell differentiation was measured by their capacity to secrete effector cytokines (interferon-gamma) and kill breast cancer cells. Specificity was measured using peptides derived from defined breast cancer antigens. RESULTS: We found that DCs loaded with killed breast cancer cells can prime naive CD8+ T cells to differentiate into effector cytotoxic T lymphocytes (CTLs). Importantly, these CTLs primed by DCs loaded with killed HLA-A*0201- breast cancer cells can kill HLA-A*0201+ breast cancer cells. Among the tumor specific CTLs, we found that CTLs specific for HLA-A2 restricted peptides derived from three well known shared breast tumor antigens, namely cyclin B1, MUC-1 and survivin. CONCLUSION: This ability of DCs loaded with killed allogeneic breast cancer cells to elicit multiantigen specific immunity supports their use as vaccines in patients with breast cancer. PMID- 17129373 TI - A gene expression signature associated with survival in metastatic melanoma. AB - BACKGROUND: Current clinical and histopathological criteria used to define the prognosis of melanoma patients are inadequate for accurate prediction of clinical outcome. We investigated whether genome screening by means of high-throughput gene microarray might provide clinically useful information on patient survival. METHODS: Forty-three tumor tissues from 38 patients with stage III and stage IV melanoma were profiled with a 17,500 element cDNA microarray. Expression data were analyzed using significance analysis of microarrays (SAM) to identify genes associated with patient survival, and supervised principal components (SPC) to determine survival prediction. RESULTS: SAM analysis revealed a set of 80 probes, corresponding to 70 genes, associated with survival, i.e. 45 probes characterizing longer and 35 shorter survival times, respectively. These transcripts were included in a survival prediction model designed using SPC and cross-validation which allowed identifying 30 predicting probes out of the 80 associated with survival. CONCLUSION: The longer-survival group of genes included those expressed in immune cells, both innate and acquired, confirming the interplay between immunological mechanisms and the natural history of melanoma. Genes linked to immune cells were totally lacking in the poor-survival group, which was instead associated with a number of genes related to highly proliferative and invasive tumor cells. PMID- 17129374 TI - Induction of multiple matrix metalloproteinases in human dermal and synovial fibroblasts by Staphylococcus aureus: implications in the pathogenesis of septic arthritis and other soft tissue infections. AB - Infections of body tissue by Staphylococcus aureus are quickly followed by degradation of connective tissue. Patients with rheumatoid arthritis are more prone to S. aureus-mediated septic arthritis. Various types of collagen form the major structural matrix of different connective tissues of the body. These different collagens are degraded by specific matrix metalloproteinases (MMPs) produced by fibroblasts, other connective tissue cells, and inflammatory cells that are induced by interleukin-1 (IL-1) and tumor necrosis factor (TNF). To determine the host's contribution in the joint destruction of S. aureus-mediated septic arthritis, we analyzed the MMP expression profile in human dermal and synovial fibroblasts upon exposure to culture supernatant and whole cell lysates of S. aureus. Human dermal and synovial fibroblasts treated with cell lysate and filtered culture supernatants had significantly enhanced expression of MMP-1, MMP 2, MMP-3, MMP-7, MMP-10, and MMP-11 compared with the untreated controls (p < 0.05). In the S. aureus culture supernatant, the MMP induction activity was identified to be within the molecular-weight range of 30 to >50 kDa. The MMP expression profile was similar in fibroblasts exposed to a combination of IL 1/TNF. mRNA levels of several genes of the mitogen-activated protein kinase (MAPK) signal transduction pathway were significantly elevated in fibroblasts treated with S. aureus cell lysate and culture supernatant. Also, tyrosine phosphorylation was significantly higher in fibroblasts treated with S. aureus components. Tyrosine phosphorylation and MAPK gene expression patterns were similar in fibroblasts treated with a combination of IL-1/TNF and S. aureus. Mutants lacking staphylococcal accessory regulator (Sar) and accessory gene regulator (Agr), which cause significantly less severe septic arthritis in murine models, were able to induce expression of several MMP mRNA comparable with that of their isogenic parent strain but induced notably higher levels of tissue inhibitors of metalloproteinases (TIMPs). To our knowledge, this is the first report of induction of multiple MMP/TIMP expression from human dermal and synovial fibroblasts upon S. aureus treatment. We propose that host-derived MMPs contribute to the progressive joint destruction observed in S. aureus-mediated septic arthritis. PMID- 17129375 TI - Abnormal insulin-like growth factor 1 signaling in human osteoarthritic subchondral bone osteoblasts. AB - Insulin-like growth factor (IGF)-1 is a key factor in bone homeostasis and could be involved in bone tissue sclerosis as observed in osteoarthritis (OA). Here, we compare the key signaling pathways triggered in response to IGF-1 stimulation between normal and OA osteoblasts (Obs). Primary Obs were prepared from the subchondral bone of tibial plateaus of OA patients undergoing knee replacement or from normal individuals at autopsy. Phenotypic characterization of Obs was evaluated with alkaline phosphatase and osteocalcin release. The effect of IGF-1 on cell proliferation, alkaline phosphatase and collagen synthesis was evaluated in the presence or not of 50 ng/ml IGF-1, whereas signaling was studied with proteins separated by SDS-PAGE before western blot analysis. We also used immunoprecipitation followed by western blot analysis to detect interactions between key IGF-1 signaling elements. IGF-1 receptor (IGF-1R), Shc, Grb2, insulin receptor substrate (IRS)-1, and p42/44 mitogen-activated protein kinase (MAPK) levels were similar in normal and OA Obs in the presence or absence of IGF-1. After IGF-1 stimulation, the phosphorylation of IGF-1R in normal and OA Obs was similar; however, the phosphorylation of IRS-1 was reduced in OA Ob. In addition, the PI3K pathway was activated similarly in normal and OA Obs while that for p42/44 MAPK was higher in OA Obs compared to normal. p42/44 MAPK can be triggered via an IRS-1/Syp or Grb2/Shc interaction. Interestingly, Syp was poorly phosphorylated under basal conditions in normal Obs and was rapidly phosphorylated upon IGF-1 stimulation, yet Syp showed a poor interaction with IRS 1. In contrast, Syp was highly phosphorylated in OA Obs and its interaction with IRS-1 was very strong initially, yet rapidly dropped with IGF-1 treatments. The interaction of Grb2 with IRS-1 progressively increased in response to IGF-1 in OA Obs whereas this was absent in normal Ob. IGF-1 stimulation altered alkaline phosphatase in Ob, an effect reduced in the presence of PD98059, an inhibitor of p42/44 MAPK signaling, whereas neither IGF-1 nor PD98059 had any significant effect on collagen synthesis. In contrast, cell proliferation was higher in OA Obs compared to normal under basal conditions, and IGF-1 stimulated more cell proliferation in OA Obs than in normal Ob, an effect totally dependent on p42/44 MAPK activiy. The altered response of OA Obs to IGF-1 may be due to abnormal IGF 1 signaling in these cells. This is mostly linked with abnormal IRS-1/Syp and IRS 1/Grb2 interaction in these cells. PMID- 17129376 TI - A psychological approach to providing self-management education for people with type 2 diabetes: the Diabetes Manual. AB - BACKGROUND: The objectives of this study were twofold (i) to develop the Diabetes Manual, a self-management educational intervention aimed at improving biomedical and psychosocial outcomes (ii) to produce early phase evidence relating to validity and clinical feasibility to inform future research and systematic reviews. METHODS: Using the UK Medical Research Council's complex intervention framework, the Diabetes Manual and associated self management interventions were developed through pre-clinical, and phase I evaluation phases guided by adult learning and self-efficacy theories, clinical feasibility and health policy protocols. A qualitative needs assessment and an RCT contributed data to the pre clinical phase. Phase I incorporated intervention development informed by the pre clinical phase and a feasibility survey. RESULTS: The pre-clinical and phase I studies resulted in the production in the Diabetes Manual programme for trial evaluation as delivered within routine primary care consultations. CONCLUSION: This complex intervention shows early feasibility and face validity for both diabetes health professionals and people with diabetes. Randomised trial will determine effectiveness against clinical and psychological outcomes. Further study of some component parts, delivered in alternative combinations, is recommended. PMID- 17129377 TI - The demand control model and circadian saliva cortisol variations in a Swedish population based sample (The PART study). AB - BACKGROUND: Previous studies of the relationship between job strain and blood or saliva cortisol levels have been small and based on selected occupational groups. Our aim was to examine the association between job strain and saliva cortisol levels in a population-based study in which a number of potential confounders could be adjusted for. METHODS: The material derives from a population-based study in Stockholm on mental health and its potential determinants. Two data collections were performed three years apart with more than 8500 subjects responding to a questionnaire in both waves. In this paper our analyses are based on 529 individuals who held a job, participated in both waves as well as in an interview linked to the second wave. They gave saliva samples at awakening, half an hour later, at lunchtime and before going to bed on a weekday in close connection with the interview. Job control and job demands were assessed from the questionnaire in the second wave. Mixed models were used to analyse the association between the demand control model and saliva cortisol. RESULTS: Women in low strain jobs (high control and low demands) had significantly lower cortisol levels half an hour after awakening than women in high strain (low control and high demands), active (high control and high demands) or passive jobs (low control and low demands). There were no significant differences between the groups during other parts of the day and furthermore there was no difference between the job strain, active and passive groups. For men, no differences were found between demand control groups. CONCLUSION: This population-based study, on a relatively large sample, weakly support the hypothesis that the demand control model is associated with saliva cortisol concentrations. PMID- 17129378 TI - Novel self-epitopes derived from aggrecan, fibrillin, and matrix metalloproteinase-3 drive distinct autoreactive T-cell responses in juvenile idiopathic arthritis and in health. AB - Juvenile idiopathic arthritis (JIA) is a heterogeneous autoimmune disease characterized by chronic joint inflammation. Knowing which antigens drive the autoreactive T-cell response in JIA is crucial for the understanding of disease pathogenesis and additionally may provide targets for antigen-specific immune therapy. In this study, we tested 9 self-peptides derived from joint-related autoantigens for T-cell recognition (T-cell proliferative responses and cytokine production) in 36 JIA patients and 15 healthy controls. Positive T-cell proliferative responses (stimulation index > or =2) to one or more peptides were detected in peripheral blood mononuclear cells (PBMC) of 69% of JIA patients irrespective of major histocompatibility complex (MHC) genotype. The peptides derived from aggrecan, fibrillin, and matrix metalloproteinase (MMP)-3 yielded the highest frequency of T-cell proliferative responses in JIA patients. In both the oligoarticular and polyarticular subtypes of JIA, the aggrecan peptide induced T-cell proliferative responses that were inversely related with disease duration. The fibrillin peptide, to our knowledge, is the first identified autoantigen that is primarily recognized in polyarticular JIA patients. Finally, the epitope derived from MMP-3 elicited immune responses in both subtypes of JIA and in healthy controls. Cytokine production in short-term peptide-specific T cell lines revealed production of interferon-gamma (aggrecan/MMP-3) and interleukin (IL)-17 (aggrecan) and inhibition of IL-10 production (aggrecan). Here, we have identified a triplet of self-epitopes, each with distinct patterns of T-cell recognition in JIA patients. Additional experiments need to be performed to explore their qualities and role in disease pathogenesis in further detail. PMID- 17129379 TI - Early onset torsion dystonia (Oppenheim's dystonia). AB - Early onset torsion dystonia (EOTD) is a rare movement disorder characterized by involuntary, repetitive, sustained muscle contractions or postures involving one or more sites of the body. A US study estimated the prevalence at approximately 1 in 30,000. The estimated prevalence in the general population of Europe seems to be lower, ranging from 1 in 330,000 to 1 in 200,000, although precise numbers are currently not available. The estimated prevalence in the Ashkenazi Jewish population is approximately five to ten times higher, due to a founder mutation. Symptoms of EOTD typically develop first in an arm or leg in middle to late childhood and progress in approximately 30% of patients to other body regions (generalized dystonia) within about five years. Distribution and severity of symptoms vary widely between affected individuals. The majority of cases from various ethnic groups are caused by an autosomal dominantly inherited deletion of 3 bp (GAG) in the DYT1 gene on chromosome 9q34. This gene encodes a protein named torsinA, which is presumed to act as a chaperone protein associated with the endoplasmic reticulum and the nuclear envelope. It may interact with the dopamine transporter and participate in intracellular trafficking, although its precise function within the cell remains to be determined. Molecular genetic diagnostic and genetic counseling is recommended for individuals with age of onset below 26 years, and may also be considered in those with onset after 26 years having a relative with typical early onset dystonia. Treatment options include botulinum toxin injections for focal symptoms, pharmacological therapy such as anticholinergics (most commonly trihexiphenydil) for generalized dystonia and surgical approaches such as deep brain stimulation of the internal globus pallidus or intrathecal baclofen application in severe cases. All patients have normal cognitive function, and despite a high rate of generalization of dystonia, 75% of those patients are able to maintain ambulation and independence, and therefore a comparatively good quality of life, with modern treatment modalities. PMID- 17129380 TI - A pilot Internet "value of health" panel: recruitment, participation and compliance. AB - OBJECTIVES: To pilot using a panel of members of the public to provide preference data via the Internet METHODS: A stratified random sample of members of the general public was recruited and familiarized with the standard gamble procedure using an Internet based tool. Health states were periodically presented in "sets" corresponding to different conditions, during the study. The following were described: Recruitment (proportion of people approached who were trained); Participation (a) the proportion of people trained who provided any preferences and (b) the proportion of panel members who contributed to each "set" of values; and Compliance (the proportion, per participant, of preference tasks which were completed). The influence of covariates on these outcomes was investigated using univariate and multivariate analyses. RESULTS: A panel of 112 people was recruited. 23% of those approached (n = 5,320) responded to the invitation, and 24% of respondents (n = 1,215) were willing to participate (net = 5.5%). However, eventual recruitment rates, following training, were low (2.1% of those approached). Recruitment from areas of high socioeconomic deprivation and among ethnic minority communities was low. Eighteen sets of health state descriptions were considered over 14 months. 74% of panel members carried out at least one valuation task. People from areas of higher socioeconomic deprivation and unmarried people were less likely to participate. An average of 41% of panel members expressed preferences on each set of descriptions. Compliance ranged from 3% to 100%. CONCLUSION: It is feasible to establish a panel of members of the general public to express preferences on a wide range of health state descriptions using the Internet, although differential recruitment and attrition are important challenges. Particular attention to recruitment and retention in areas of high socioeconomic deprivation and among ethnic minority communities is necessary. Nevertheless, the panel approach to preference measurement using the Internet offers the potential to provide specific utility data in a responsive manner for use in economic evaluations and to address some of the outstanding methodological uncertainties in this field. PMID- 17129381 TI - Relationship between parental estimate and an objective measure of child television watching. AB - Many young children have televisions in their bedrooms, which may influence the relationship between parental estimate and objective measures of child television usage/week. Parental estimates of child television time of eighty 4-7 year old children (6.0 +/- 1.2 years) at the 75th BMI percentile or greater (90.8 +/- 6.8 BMI percentile) were compared to an objective measure of television time obtained from TV Allowance devices attached to every television in the home over a three week period. Results showed that parents overestimate their child's television time compared to an objective measure when no television is present in the bedroom by 4 hours/week (25.4 +/- 11.5 vs. 21.4 +/- 9.1) in comparison to underestimating television time by over 3 hours/week (26.5 +/- 17.2 vs. 29.8 +/- 14.4) when the child has a television in their bedroom (p = 0.02). Children with a television in their bedroom spend more objectively measured hours in television time than children without a television in their bedroom (29.8 +/- 14.2 versus 21.4 +/- 9.1, p = 0.003). Research on child television watching should take into account television watching in bedrooms, since it may not be adequately assessed by parental estimates. PMID- 17129382 TI - Small interfering RNA targeted to stem-loop II of the 5' untranslated region effectively inhibits expression of six HCV genotypes. AB - BACKGROUND: The antiviral action of interferon alpha targets the 5' untranslated region (UTR) used by hepatitis C virus (HCV) to translate protein by an internal ribosome entry site (IRES) mechanism. Although this sequence is highly conserved among different clinical strains, approximately half of chronically infected hepatitis C patients do not respond to interferon therapy. Therefore, development of small interfering RNA (siRNA) targeted to the 5'UTR to inhibit IRES mediated translation may represent an alternative approach that could circumvent the problem of interferon resistance. RESULTS: Four different plasmid constructs were prepared for intracellular delivery of siRNAs targeting the stem loop II-III of HCV 5' UTR. The effect of siRNA production on IRES mediated translation was investigated using chimeric clones between the gene for green fluorescence protein (GFP) and IRES sequences of six different HCV genotypes. The siRNA targeted to stem loop II effectively mediated degradation of HCV IRES mRNA and inhibited GFP expression in the case of six different HCV genotypes, where as siRNAs targeted to stem loop III did not. Furthermore, intracytoplasmic expression of siRNA into transfected Huh-7 cells efficiently degraded HCV genomic RNA and inhibited core protein expression from infectious full-length infectious clones HCV 1a and HCV 1b strains. CONCLUSION: These in vitro studies suggest that siRNA targeted to stem-loop II is highly effective inhibiting IRES mediated translation of the major genotypes of HCV. Stem-loop II siRNA may be a good target for developing an intracellular immunization strategy based antiviral therapy to inhibit hepatitis C virus strains that are not inhibited by interferon. PMID- 17129383 TI - Molecular subtypes of breast cancer in relation to paclitaxel response and outcomes in women with metastatic disease: results from CALGB 9342. AB - INTRODUCTION: The response to paclitaxel varies widely in metastatic breast cancer. We analyzed data from CALGB 9342, which tested three doses of paclitaxel in women with advanced disease, to determine whether response and outcomes differed according to HER2, hormone receptor, and p53 status. METHODS: Among 474 women randomly assigned to paclitaxel at a dose of 175, 210, or 250 mg/m2, adequate primary tumor tissue was available from 175. Immunohistochemistry with two antibodies and fluorescence in situ hybridization were performed to evaluate HER2 status; p53 status was determined by immunohistochemistry and sequencing. Hormone receptor status was obtained from pathology reports. RESULTS: Objective response rate was not associated with HER2 or p53 status. There was a trend toward a shorter median time to treatment failure among women with HER2-positive tumors (2.3 versus 4.2 months; P = 0.067). HER2 status was not related to overall survival (OS). Hormone receptor expression was not associated with differences in response but was associated with longer OS (P = 0.003). In contrast, women with p53 over-expression had significantly shorter OS than those without p53 over expression (11.5 versus 14.4 months; P = 0.002). In addition, triple negative tumors were more frequent in African-American than in Caucasian patients, and were associated with a significant reduction in OS (8.7 versus 12.9 months; P = 0.008). CONCLUSION: None of the biomarkers was predictive of treatment response in women with metastatic breast cancer; however, survival differed according to hormone receptor and p53 status. Triple negative tumors were more frequent in African-American patients and were associated with a shorter survival. PMID- 17129384 TI - Did the last common ancestor have a biological membrane? AB - All theories about the origin and evolution of membrane bound cells necessarily have to cope with the nature of the last common ancestor of cellular life. One of the most important aspect of this ancestor, whether it had a closed biological membrane or not, has recently been intensely debated. Having a consensus about it would be an important step towards an eventual (though probably still remote) synthesis of the best elements of the current multitude of cell evolution models. Here I analyse the structural and functional conservation of the few universally distributed proteins that were undoubtedly present in the last common ancestor and that carry out membrane-associated functions. These include the SecY subunit of the protein-conducting channel, the signal recognition particle, the signal recognition particle receptor, the signal peptidase, and the proton ATPase. The conserved structural and functional aspects of these proteins indicate that the last common ancestor was associated with a hydrophobic layer with two hydrophilic sides (an inside and an outside) that had a full-fledged and asymmetric protein insertion and translocation machinery and served as a permeability barrier for protons and other small molecules. It is difficult to escape the conclusion that the last common ancestor had a closed biological membrane from which all cellular membranes evolved. PMID- 17129385 TI - Relationship between tonic inhibitory currents and phasic inhibitory activity in the spinal cord lamina II region of adult mice. AB - Phasic and tonic inhibitions are two types of inhibitory activities involved in inhibitory processing in the CNS. In the spinal cord dorsal horn, phasic inhibition is mediated by both GABAergic and glycinergic inhibitory postsynaptic currents. In contrast to phasic inhibitory currents, using patch-clamp recording technique on spinal cord slices prepared from adult mice we revealed that tonic inhibitory currents were mediated by GABAA receptors but not by glycine receptors in dorsal horn lamina II region. We found that there was a linear relationship (r = 0.85) between the amplitude of tonic inhibitory currents and the frequency of GABAergic inhibitory postsynaptic currents. Analysis of charge transfer showed that the charges carried by tonic inhibitory currents were about 6 times of charges carried by phasic inhibitory currents. The prominent charge transfer by tonic inhibitory currents and their synaptic activity dependency suggest a significant role of tonic inhibition in sensory processing. PMID- 17129386 TI - Integrated siRNA design based on surveying of features associated with high RNAi effectiveness. AB - BACKGROUND: Short interfering RNAs have allowed the development of clean and easily regulated methods for disruption of gene expression. However, while these methods continue to grow in popularity, designing effective siRNA experiments can be challenging. The various existing siRNA design guidelines suffer from two problems: they differ considerably from each other, and they produce high levels of false-positive predictions when tested on data of independent origins. RESULTS: Using a distinctly large set of siRNA efficacy data assembled from a vast diversity of origins (the siRecords data, containing records of 3,277 siRNA experiments targeting 1,518 genes, derived from 1,417 independent studies), we conducted extensive analyses of all known features that have been implicated in increasing RNAi effectiveness. A number of features having positive impacts on siRNA efficacy were identified. By performing quantitative analyses on cooperative effects among these features, then applying a disjunctive rule merging (DRM) algorithm, we developed a bundle of siRNA design rule sets with the false positive problem well curbed. A comparison with 15 online siRNA design tools indicated that some of the rule sets we developed surpassed all of these design tools commonly used in siRNA design practice in positive predictive values (PPVs). CONCLUSION: The availability of the large and diverse siRNA dataset from siRecords and the approach we describe in this report have allowed the development of highly effective and generally applicable siRNA design rule sets. Together with ever improving RNAi lab techniques, these design rule sets are expected to make siRNAs a more useful tool for molecular genetics, functional genomics, and drug discovery studies. PMID- 17129387 TI - CcpA affects expression of the groESL and dnaK operons in Lactobacillus plantarum. AB - BACKGROUND: Lactic acid bacteria (LAB) are widely used in food industry and their growth performance is important for the quality of the fermented product. During industrial processes changes in temperature may represent an environmental stress to be overcome by starters and non-starters LAB. Studies on adaptation to heat shock have shown the involvement of the chaperon system-proteins in various gram positive bacteria. The corresponding operons, namely the dnaK and groESL operons, are controlled by a negative mechanism involving the HrcA repressor protein binding to the cis acting element CIRCE. RESULTS: We studied adaptation to heat shock in the lactic acid bacterium Lactobacillus plantarum. The LM3-2 strain, carrying a null mutation in the ccpA gene, encoding the catabolite control protein A (CcpA), showed a lower percent of survival to high temperature with respect to the LM3 wild type strain. Among proteins differentially expressed in the two strains, the GroES chaperon was more abundant in the wild type strain compared to the mutant strain under standard growth conditions. Transcriptional studies showed that class I heat shock operons were differentially expressed upon heat shock in both strains. Indeed, the dnaK and groESL operons were induced about two times more in the LM3 strain compared to the LM3-2 strain. Analysis of the regulatory region of the two operons showed the presence of cre sequences, putative binding sites for the CcpA protein. CONCLUSION: The L. plantarum dnaK and groESL operons are characterized by the presence of the cis acting sequence CIRCE in the promoter region, suggesting a negative regulation by the HrcA/CIRCE system, which is a common type of control among the class I heat shock operons of gram-positive bacteria. We found an additional system of regulation, based on a positive control exerted by the CcpA protein, which would interact with cre sequences present in the regulatory region of the dnaK and groESL operons. The absence of the CcpA protein results in a lower induction of the chaperon coding operons, with a consequent lower percent of survival of the LM3-2 mutant strain population with respect to the wild type when challenged with a heat insult. PMID- 17129388 TI - Early changes of CD4-positive lymphocytes and NK cells in patients with severe Gram-negative sepsis. AB - INTRODUCTION: Our aim was to define early changes of lymphocytes and of NK cells in severe sepsis and to correlate them with serum levels of soluble triggering receptor expressed on myeloid cells-1 (sTREM-1). METHODS: Blood was sampled from 49 patients with proven highly suspected infection by Gram-negative pathogens, within 12 hours of the advent of severe sepsis, and was also sampled from six healthy volunteers. White blood cells were targeted with monoclonal antibodies and were analyzed by flow cytometry. The concentrations of sTREM-1 were estimated by ELISA. RESULTS: The presence of CD3/CD4 cells was significantly lower (P < 0.0001) and that of NK cells significantly higher among patients with sepsis compared with controls (P = 0.011). The proportions (median +/- standard error) of ANNEXIN-V/CD4/CD3-positive cells, of ANNEXIN-V/CD8/CD3-positive cells and of ANNEXIN-V/CD14-positive cells of the patient population were 7.41 +/- 2.26%, 7.69 +/- 3.42% and 1.96 +/- 4.22%, respectively. Patients with NK cells >20% survived longer compared with those patients with NK cells < or =20% (P = 0.041), and patients with sTREM-1 concentrations >180 pg/ml survived longer compared with those patients with sTREM-1 concentrations < or =180 pg/ml (P = 0.042). A negative correlation was found between the percentages of ANNEXIN-V/CD4/CD3 positive cells and of CD3/CD4 cells (rs = -0.305, P = 0.049), and a positive correlation was found between the serum sTREM-1 concentration and the percentage of NK cells (rs = +0.395, P = 0.014). NK cells isolated from two healthy volunteers released sTREM-1 upon triggering with endotoxins. CONCLUSION: Early severe sepsis is characterized by CD4-lymphopenia and increased NK cells, providing a survival benefit for the septic patient at percentages >20%. The survival benefit resulting from elevated NK cells might be connected to elevated serum levels of sTREM-1. PMID- 17129389 TI - Randomised controlled single-blind study of conventional versus depot mydriatic drug delivery prior to cataract surgery. AB - BACKGROUND: A prerequisite for safe cataract surgery is an adequately dilated pupil. The authors conducted a trial to assess the efficacy (in terms of pupil diameter) of a depot method of pre-operative pupil dilatation, as compared with repeated instillations of drops (which is time-consuming for the nursing staff and uncomfortable for the patient). METHODS: A prospective randomised masked trial was conducted comprising 130 patients with no significant ocular history undergoing elective clear corneal phacoemulsification. 65 patients had mydriatic drops (Tropicamide 1%, Phenylephrine 2.5%, Diclofenac sodium 0.1%) instilled prior to surgery, 65 had a wick soaked in the same drop mixture placed in the inferior fornix. Horizontal pupil diameters were recorded on a millimetre scale immediately prior to surgery. RESULTS: There was no significant difference in pupil size between the two groups (p = 0.255, Student's t-test). CONCLUSION: There was no significant difference between the mydriasis obtained with the depot system compared with conventional drop application. Use of a depot mydriatic delivery system appears to be a safe and efficient method of drug delivery. TRIAL REGISTRATION: International Standard Randomised Controlled Trial Number Register ISRCTN78047760. PMID- 17129390 TI - Laryngocele: a rare complication of surgical tracheostomy. AB - BACKGROUND: A laryngocele is usually a cystic dilatation of the laryngeal saccule. The etiology behind its occurrence is still unclear, but congenital and acquired factors have been implicated in its development. CASE PRESENTATION: We present a rare case of laryngocele occurring in a 77-year-old Caucasian woman. The patient presented with one month history of altered voice, no other associated symptoms were reported. The medical history of the patient included respiratory failure secondary to childhood polio at the age of ten; the airway management included a surgical tracheostomy. Flexible naso-laryngoscopy revealed a soft mass arising from the posterior pharyngeal wall obscuring the view of the posterior commissure and vocal folds. The shape of the mass altered with respiration and on performing valsalva maneuver. A plain lateral neck radiograph revealed a large air filled sac originating from the laryngeal cartilages and extending along the posterior pharyngeal wall. The patient was then treated by endoscopic laser marsupialization and reviewed annually.We discuss the complications of tracheostomy and the pathophysiology of laryngoceles and in particular the likely aetiological factors in this case. CONCLUSION: A laryngocele presenting in a female patient with tracheostomy is extremely rare and has not been to date reported in the world literature. A local mechanical condition may be the determinant factor in the pathogenesis of the disease. PMID- 17129391 TI - Hepatocellular proliferation in response to agonists of peroxisome proliferator activated receptor alpha: a role for Kupffer cells? AB - BACKGROUND: It has been proposed that PPARalpha agonists stimulate Kupffer cells in rodents which in turn, release mitogenic factors leading to hepatic hyperplasia, and eventually cancer. However, Kupffer cells do not express PPARalpha receptors, and PPARalpha agonists stimulate hepatocellular proliferation in both TNFalpha- and TNFalpha receptor-null mice, casting doubt on the involvement of Kupffer cells in the mitogenic response to PPARalpha agonists. This study was therefore designed to investigate whether the PPARalpha agonist PFOA and the Kupffer cell inhibitor methylpalmitate produce opposing effects on hepatocellular proliferation and Kupffer cell activity in vivo, in a manner that would implicate these cells in the mitogenic effects of PPARalpha agonists. METHODS: Male Sprague-Dawley rats were treated intravenously via the tail vein with methylpalmitate 24 hrs prior to perfluorooctanoic acid (PFOA), and were sacrificed 24 hrs later, one hr after an intraperitoneal injection of bromodeoxyuridine (BrdU). Sera were analyzed for TNFalpha and IL-1beta. Liver sections were stained immunohistochemically and quantified for BrdU incorporated into DNA. RESULTS: Data show that PFOA remarkably stimulated hepatocellular proliferation in the absence of significant changes in the serum levels of either TNFalpha or IL-1beta. In addition, methylpalmitate did not alter the levels of these mitogens in PFOA-treated animals, despite the fact that it significantly blocked the hepatocellular proliferative effect of PFOA. Correlation between hepatocellular proliferation and serum levels of TNFalpha or IL-1beta was extremely poor. CONCLUSION: It is unlikely that mechanisms involving Kupffer cells play an eminent role in the hepatic hyperplasia, and consequently hepatocarcinogenicity attributed to PPARalpha agonists. This conclusion is based on the above mentioned published data and the current findings showing animals treated with PFOA alone or in combination with methylpalmitate to have similar levels of serum TNFalpha and IL-1beta, which are reliable indicators of Kupffer cell activity, despite a remarkable difference in hepatocellular proliferation. PMID- 17129392 TI - Large B-cell lymphoma presenting as acute abdominal pain and spontaneous splenic rupture; a case report and review of relevant literature. AB - BACKGROUND: Spontaneous rupture of the spleen is an uncommon dramatic abdominal emergency that requires immediate diagnosis and prompt surgical treatment to ensure the patients survival. Infections have been cited in most cases involving splenic rupture but are rare in hematological malignancies despite frequent involvement of the spleen. METHODS AND MATERIALS: We present a case of a splenic rupture caused by infiltration of B-cell lymphoma. A 43 year old gentleman presented with a 1 day h/o left upper quadrant pain; nausea and vomiting for 2 days with associated dizziness and anorexia. The CT showed abnormal spleen 20 x 11 cm with free fluid in the abdomen and enlarged retroperitoneal LNs. The patient underwent a splenectomy after initial resuscitation and the operative finding was that of a massively enlarged spleen with areas of tumor extruding through the splenic capsule. RESULT AND CONCLUSION: Although the spleen is often involved in hematological malignancies, splenic rupture is an infrequent occurrence. In a recent literature review 136 cases were of splenic rupture secondary to hematological malignancy were identified. Acute leukemia and non Hodgkin lymphoma were the frequent causes followed by chronic myelogenous leukemia. Male sex, adulthood, severe splenomegaly and cytoreductive chemotherapy were factors more often associated with splenic rupture. Emergency splenectomy remains the cornerstone treatment for splenic rupture. We present a case report of a "spontaneous splenic rupture" and discuss the presentation, etiology and treatment options along with discussion of relevant literature. PMID- 17129393 TI - Even more suicide attempts in clinical trials with paroxetine randomised against placebo. AB - BACKGROUND: Following our previous publication we have received critical comments to our conclusions as well as new data that are strengthening our findings. RESULTS: With the new data, 11 suicide attempts among patients on paroxetine against 1 among patients on placebo, we found with a Bayesian technique that the posterior probability that medication with paroxetine is associated with an increased intensity per year of a suicide attempt is from 0.98 to 0.99, depending on the prior. We found that the comment to our article by GSK representatives contained errors, misunderstanding and unwillingness to accept Bayesian principles in the analysis of clinical trials. CONCLUSION: We were in our previous publication, with preliminary data and a Bayesian approach, able to raise a concern that suicide attempts might be connected with the use of paroxetine. This suspicion has now been confirmed. PMID- 17129394 TI - Inhibition of PKC activity blocks the increase of ETB receptor expression in cerebral arteries. AB - BACKGROUND: Previous studies have shown that there is a time-dependent upregulation of contractile endothelin B (ETB) receptors in middle cerebral arteries (MCA) after organ culture. This upregulation is dependent on mitogen activated protein kinases and possibly protein kinase C (PKC). The aim of this study was to examine the effect of PKC inhibitors with different profiles on the upregulation of contractile ETB receptors in rat MCA. Artery segments were incubated for 24 hours at 37 degrees C. To investigate involvement of PKC, inhibitors were added to the medium before incubation. The contractile endothelin mediated responses were measured and real-time PCR was used to detect endothelin receptor mRNA levels. Furthermore, immunohistochemistry was used to demonstrate the ETB receptor protein distribution in the MCA and Western blot to measure which of the PKC subtypes that were affected by the inhibitors. RESULTS: The PKC inhibitors bisindolylmaleimide I, Ro-32-0432 and PKC inhibitor 20-28 attenuated the ETB receptor mediated contractions. Furthermore, Ro-32-0432 and bisindolylmaleimide I decreased ETB receptor mRNA levels while PKC inhibitor 20 28 reduced the amount of receptor protein on smooth muscle cells. PKC inhibitor 20-28 also decreased the protein levels of the five PKC subtypes studied (alpha, betaI, gamma, delta and epsilon). CONCLUSION: The results show that PKC inhibitors are able to decrease the ETB receptor contraction and expression in MCA smooth muscle cells following organ culture. The PKC inhibitor 20-28 affects the protein levels, while Ro-32-0432 and bisindolylmaleimide I affect the mRNA levels, suggesting differences in activity profile. Since ETB receptor upregulation is seen in cerebral ischemia, the results of the present study provide a way to interfere with the vascular involvement in cerebral ischemia. PMID- 17129395 TI - Specific cpb copies within the Leishmania donovani complex: evolutionary interpretations and potential clinical implications in humans. AB - Leishmania infantum and Leishmania donovani both pertain to the L. (L.) donovani complex and are responsible for visceral leishmaniasis. To explore the L. donovani complex, we focused our study on cysteine protease B (cpb) and especially on 2 cpb copies: cpbE and cpbF. We selected cpb genes because of their phylogenetic interest and host-parasite interaction involvement. Sequencing these 2 copies revealed (i) that cpbE is specific to L. infantum and cpbF is specific to L. donovani and (ii) that these 2 copies are different in length and sequence. Phylogenetic analysis and protein predictions were carried out in order to compare these copies (i) with other trypanosomatid cpb, especially L. mexicana, and (ii) within the L. donovani complex. Our results revealed patterns specific to the L. donovani complex such as the COOH-terminal extension, potential epitopes and N-glycosylation sites. Moreover, phylogenetic analysis revealed different levels of polymorphism between L. infantum and L. donovani and confirmed the ancestral status of the latter. L. infantum has a shorter sequence and a deleted sequence responsible for modifications in protein conformation and catalytic triad. Considering the clinical aspect, L. infantum dermotropic strains appeared more polymorphic than L. infantum viscerotropic strains. PMID- 17129396 TI - Socio-economic and psychological correlates of suicidality among Hong Kong working-age adults: results from a population-based survey. AB - BACKGROUND: The global toll of suicide is estimated to be one million lives per year, which exceeded the number of deaths by homicide and war combined. A key step to suicide prevention is to prevent less serious suicidal behaviour to preclude more lethal outcomes. Although 61% of the world's suicides take place in Asia and the suicide rates among middle age groups have been increasing since the economic crisis in many Asian countries, population-based studies of suicidal behaviour among working-age adults in non-western communities are scarce. METHOD: Data from a population-based survey with 2015 participants were used to estimate the prevalence of suicidal ideation and behaviour among the working-age population in Hong Kong, and to study the associated socio-economic and psychological correlates. We focused particularly on potential modulating factors between life-event-related factors and suicidal ideation. RESULTS: Six per cent of the Hong Kong population aged 20-59 years considered suicide in the past year, while 1.4% attempted suicide. Hopelessness, reasons for living, and reluctance to seek help from family and friends had direct association with past-year suicidal ideation. Reasons for living were found to moderate the effect of perceived stress on suicidal ideation. CONCLUSIONS: Suicidality is a multi-faceted problem that calls for a multi-sectored, multi-layered approach to prevention. Prevention programmes can work on modulating factors such as reasons for living to reduce suicidal risk in working-age adults. PMID- 17129397 TI - Chest wall motion after thixotropy conditioning of inspiratory muscles in healthy humans. AB - Inspiratory muscle conditioning at a lower or higher lung volume based on the principles of muscle thixotropy causes acute changes in end-expiratory chest wall and lung volumes. The present study aimed to demonstrate the time course of effects of this conditioning on both end-expiratory chest wall volume and thoracoabdominal synchrony. We measured chest wall motion with respiratory induction plethysmography at 0.5, 1, 2, 3, and 6 min after conditioning at three different lung volumes in 15 healthy men. After conditioning at total lung capacity - 20% inspiratory capacity, increases in end-expiratory chest wall volume were significant at 0.5, 1, and 2 min (P < 0.05), being most obvious at 0.5 min (Delta 0.24 +/- 0.20 liter). After conditioning at residual volume, reductions in end-expiratory chest wall volume were significant at any time point (P < 0.05), being most obvious at 0.5 min (Delta 0.16 +/- 0.08 liter). Conditioning at functional residual capacity had little effect on the volume. Spirometric inspiratory capacity at 6 min after conditioning at residual volume (2.68 +/- 0.35 liter) was higher than the baseline value (2.53 +/- 0.31 liter, P < 0.05). Reductions in the phase angle, quantified by the Konno-Mead diagram, occurred after conditioning at residual volume at any time point (P < 0.05), being most obvious at 2 min (Delta 3.47 +/- 3.02 degrees). In conclusion, there is a 6-min time course of changes in end-expiratory chest wall volume after conditioning. More synchronous motion between the rib cage and abdomen occurs after conditioning at residual volume. PMID- 17129398 TI - NSAID activated gene (NAG-1), a modulator of tumorigenesis. AB - The NSAID activated gene (NAG-1), a member of the TGF-beta superfamily, is involved in tumor progression and development. The over-expression of NAG-1 in cancer cells results in growth arrest and increase in apoptosis, suggesting that NAG-1 has anti-tumorigenic activity. This conclusion is further supported by results of experiments with transgenic mice that ubiquitously express human NAG 1. These transgenic mice are resistant to the development of intestinal tumors following treatment with azoxymethane or by introduction of a mutant APC gene. In contrast, other data suggest a pro-tumorigenic role for NAG-1, for example, high expression of NAG-1 is frequently observed in tumors. NAG-1 may be like other members of the TGF-beta superfamily, acting as a tumor suppressor in the early stages, but acting pro-tumorigenic at the later stages of tumor progression. The expression of NAG-1 can be increased by treatment with drugs and chemicals documented to prevent tumor formation and development. Most notable is the increase in NAG-1 expression by the inhibitors of cyclooxygenases that prevent human colorectal cancer development. The regulation of NAG-1 is complex, but these agents act through either p53 or EGR-1 related pathways. In addition, an increase in NAG-1 is observed in inhibition of the AKT/GSK-3beta pathway, suggesting NAG-1 alters cell survival. Thus, NAG-1 expression is regulated by tumor suppressor pathways and appears to modulate tumor progression. PMID- 17129399 TI - Silymarin modulates Cisplatin-induced oxidative stress and hepatotoxicity in rats. AB - Cisplatin (CDDP) is a widely used anticancer drug, but at high dose, it can produce undesirable side effects such as hepatotoxicity. Because silymrin has been used to treat liver disorders, the protective effect of silymarin on CDDP induced hepatotoxicity was evaluated in rats. Hepatotoxicity was determined by changes in serum alanine aminotransferase [ALT] and aspartate aminotransferase [AST], nitric oxide [NO] levels, albumin and calcium levels, and superoxide dismutase [SOD], glutathione peroxidase [GSHPx] activities, glutathione content, malondialdehyde [MDA] and nitric oxide [NO] levels in liver tissue of rats. Male albino rats were divided into four groups, 10 rats in each. In the control group, rats were injected i.p. with 0.2 ml of propylene glycol in saline 75/25 (v/v) for 5 consecutive days [Silymarin was dissolved in 0.2 ml of propylene glycol in saline 75/25 v/v]. The second group were injected with CDDP (7.5 mg /kg, I.P.), whereas animals in the third group were i.p. injected with silymarin at a dose of 100 mg/kg/day for 5 consecutive days. The Fourth group received a daily i.p. injection of silymarin (100 mg/kg/day for 5 days) 1 hr before a single i.p. injection of CDDP (7.5 mg/kg). CDDP hepatotoxicity was manifested biochemically by an increase in serum ALT and AST, elevation of MDA and NO in liver tissues as well as a decrease in GSH and the activities of antioxidant enzymes, including SOD, GSHPx in liver tissues. In addition, marked decrease in serum NO, albumin and calcium levels were observed. Serum ALT, AST, liver NO level, MDA was found to decreased in the combination group in comparison with the CDDP group. The activities of SOD, GSHPx, GSH and serum NO were lower in CDDP group than both the control and CDDP pretreated with silymarin groups. The results obtained suggested that silymarin significantly attenuated the hepatotoxicity as an indirect target of CDDP in an animal model of CDDP-induced nephrotoxicity. PMID- 17129400 TI - Mistletoe lectin (Viscum album coloratum) modulates proliferation and cytokine expressions in murine splenocytes. AB - It is well documented that an extract of European mistletoe has a variety of biological effects, such as the stimulation of cytokine production from immune cells, and additional immunoadjuvant activities. While the European mistletoe has been studied intensively, we know less about Korean mistletoe as a therapeutic plant, especially as a possible immunomodulating drug. This study will investigated the effects of Korean mistletoe lectin (Viscum album L. var. coloratum agglutinin, VCA) on murine splenocytes to investigate whether VCA acts as an immunomodulator, which could lead to improved immune responses in these cells. The results showed that VCA inhibited cell proliferation at higher concentrations (at 1-8 ng/ml) and enhanced cell proliferation at lower concentrations (at 4-32 pg/ml). Further studies were carried out to determine if the proproliferative or anti-proliferative activity exhibited by VCA was correlated with cytokine secretion. Consequently, interferon (IFN)-gamma secretion was decreased in concanavalin A (ConA)-stimulated murine splenocytes by VCA (4-64 ng/ml), but there was no change in IL-4 levels. This suggests that VCA has the ability to modulate murine splenocyte proliferation and can possibly act on the balance of Th1/Th2 cellular immune responses. PMID- 17129401 TI - Purification and characterization of protein phosphatase 2A from petals of the tulip Tulipa gesnerina. AB - The holoenzyme of protein phosphatase (PP) from tulip petals was purified by using hydrophobic interaction, anion exchange and microcystin affinity chromatography to analyze activity towards p-nitrophenyl phosphate (p-NPP). The catalytic subunit of PP was released from its endogenous regulatory subunits by ethanol precipitation and further purified. Both preparations were characterized by immunological and biochemical approaches to be PP2A. On SDS-PAGE, the final purified holoenzyme preparation showed three protein bands estimated at 38, 65, and 75 kDa while the free catalytic subunit preparation showed only the 38 kDa protein. In both preparations, the 38 kDa protein was identified immunologically as the catalytic subunit of PP2A by using a monoclonal antibody against the PP2A catalytic subunit. The final 623- and 748- fold purified holoenzyme and the free catalytic preparations, respectively, exhibited high sensitivity to inhibition by 1 nM okadaic acid when activity was measured with p-NPP. The holoenzyme displayed higher stimulation in the presence of ammonium sulfate than the free catalytic subunit did by protamine, thereby suggesting different enzymatic behaviors. PMID- 17129402 TI - Overexpressed Derlin-1 inhibits ER expansion in the endothelial cells derived from human hepatic cavernous hemangioma. AB - Proteins that are unfolded or misfolded in the endoplasmic reticulum (ER) must be targeted for refolding or degradation to maintain the homeostasis of the ER. Derlin-1 was reportedly implicated in the retro-translocation of misfolded proteins from the ER to the cytosol for degradation. In this report, we showed that Derlin-1 was down-regulated in the endothelial cells derived from human hepatic cavernous hemangioma (CHEC) compared with other tested cells. Electron microscopy analysis showed that ER was aberrantly enlarged in CHEC cells, but not in other tested cells. When overexpressed, Derlin-1 induced the dilated ER to return normal size. This ER dynamic was associated with the activation of unfolded protein response (UPR). In CHEC cells where Derlin-1 was down-regulated, increased expression of the immunoglobulin heavy chain-binding protein (Bip) and UPR-specific splicing of X-box DNAbinding protein 1 (XBP1) mRNA were detected, as compared with that in other tested cells, indicating that UPR was activated. After Derlin-1 overexpression, the extent of UPR activation diminished, as evidenced by decreased expression of Bip, reduced amount of the spliced form of XBP1 (XBP1s), and elevated expression of the unspliced form of XBP1 (XBP1u). Taken together, these findings provide another example of a single protein being able to affect ER dynamic in mammalian cells, and an insight into the possible molecular mechanism(s). PMID- 17129403 TI - Molecular cloning, characterization and expression analysis of an ILF2 homologue from Tetraodon nigroviridis. AB - Interleukin-2 enhancer binding factor 2 (ILF2) was reported to regulate transcription of interleukin-2 (IL-2), a central cytokine in the regulation of T cell responses. This property of ILF2 was well characterized in human and mammals, but little is known in bony fish. In this paper, an ILF2 homologue was cloned and well characterized from Tetraodon nigrovirid is for the further investigation of the function of ILF2 in bony fish. The full-length Tetraodon ILF2 cDNA was 1380 bp in size and contained an open reading frame (ORF) of 1164 bp that translates into a 387 amino-acid peptide with a molecular weight of 42.9 kDa, a 5' untranslated region (UTR) of 57 bp, and a 3' UTR of 159 bp containing a poly A tail. The deduced peptide of Tetraodon ILF2 shared an overall identity of 58%~93% with other known ILF2 sequences, and contained two Nglycosylation sites, two N-myristoylation sites, one RGD cell attachment sequence, six protein kinase C phosphorylation sites, one amino-terminal RGG-rich single-stranded RNAbinding domain, and a DZF zinc-finger nucleic acid binding domain, most of which were highly conserved through species compared. Constitutive expression of Tetraodon ILF2 was observed in all tissues examined, including gill, gut, head kidney, spleen, liver, brain and heart. The highest expression was detected in heart, followed by liver, head kidney and brain. Stimulation with LPS did not significantly alter the expression of Tetraodon ILF2. Gene organization analysis showed that the Tetraodon ILF2 gene have fifteen exons, one more than other known ILF2 genes in human and mouse. Genes upand down-stream from the Tetraodon ILF2 were Rpa12, Peroxin-11b, Smad4, Snapap and Txnip homologue, which were different from that in human and mouse. PMID- 17129404 TI - Development of high-specificity antibodies against renal urate transporters using genetic immunization. AB - Recently three proteins, playing central roles in the bidirectional transport of urate in renal proximal tubules, were identified: two members of the organic anion transporter (OAT) family, OAT1 and OAT3, and a protein that designated renal urate-anion exchanger (URAT1). Antibodies against these transporters are very important for investigating their expressions and functions. With the cytokine gene as a molecular adjuvant, genetic immunization-based antibody production offers several advantages including high specificity and high recognition to the native protein compared with current methods. We fused high antigenicity fragments of the three transporters to the plasmids pBQAP-TT containing T-cell epitopes and flanking regions from tetanus toxin, respectively. Gene gun immunization with these recombinant plasmids and two other adjuvant plasmids, which express granulocyte/ macrophage colony-stimulating factor and FMS like tyrosine kinase 3 ligand, induced high level immunoglobulin G antibodies, respectively. The native corresponding proteins of URAT1, OAT1 and OAT3, in human kidney can be recognized by their specific antibodies, respectively, with Western blot analysis and immunohistochemistry. Besides, URAT1 expression in Xenopus oocytes can also be recognized by its corresponding antibody with immuno fluorescence. The successful production of the antibodies has provided an important tool for the study of UA transporters. PMID- 17129405 TI - The effect of protein expression of Streptococcus pneumoniae by blood. AB - During infection, the common respiratory tract pathogen Streptococcus pneumoniae encounters several environmental conditions, such as upper respiratory tract, lung tissue, and blood stream, etc. In this study, we examined the effects of blood on S. pneumoniae protein expression using a combination of highly sensitive 2-dimensional electrophoresis (DE) and MALDI-TOF MS and/or LC/ESI-MS/MS. A comparison of expression profiles between the growth in THY medium and THY supplemented with blood allowed us to identify 7 spots, which increased or decreased two times or more compared with the control group: tyrosyl-tRNA synthetase, lactate oxidase, glutamyl-aminopeptidase, L-lactate dehydrogenase, cysteine synthase, ribose-phosphate pyrophosphokinase, and orotate phosphoribosyltransferase. This global approach can provide a better understanding of S. pneumoniae adaptation to its human host and a clue for its pathogenicity. PMID- 17129406 TI - Screening of domain-specific target proteins of polo-like kinase 1: construction and application of centrosome/kinetochore-specific targeting peptide. AB - Mammalian polo-like kinase 1 (Plk1) acts at various stages in early and late mitosis. Plk1 localizes at the centrosome and maintains this position through mitosis. Thereafter Plk1 moves to the kinetochore and midbody region, important sites during chromosome separation and cytokinesis. The catalytic domain of Plk1 is in the Nterminus region, whereas the non-catalytic region in the Cterminus of Plk1 has a conserved motif, named the Polobox. This motif is critical for Plk localization. EGFP proteins fused with the N-terminus and C-terminus of Plk1 localize in the nucleus and centrosomes, respectively. The core sequences of the polo-box (50 amino acids) also localize in Plk1 target organelles. To screen for domainspecific target proteins of Plk1, we constructed an Nterminal domain and a tandem repeat polo-box motif, and used them as templates in a yeast two-hybrid screen. The HeLa cell cDNA library indicated several proteins including the centrosome/kinetochore components or regulators, to be characterized as positive clones. Through in vitro protein binding analyses, we confirmed an interaction between these proteins and Plk1. The data reported from this study indicate that the N- and Ctermini of Plk1 may function through recruitment and/or activation of domain-specific target proteins in dividing cells. Additionally, tandem repeats of the conserved core motif of the polo-box are sufficient for targeting and may be useful as a centrosome/kinetochore-specific targeting peptide. PMID- 17129407 TI - High level expression of a protein precursor for functional studies. AB - In vitro analyses of type I signal peptidase activities require protein precursors as substrates. Usually, these pre-proteins are expressed in vitro and cleavage of the signal sequence is followed by SDS polyacrylamide gel electrophoresis coupled with autoradiography. Radioactive amino acids have to be incorporated in the expressed protein, since the amount of the in vitro expressed protein is usually very low and processing of the signal peptide cannot be followed by SDS polyacrylamide gel electrophoresis alone. Here we describe a rapid and simple method to express large amounts of a protein precursor in E. coli. We have analyzed the effect of ionophors as well as of azide on the accumulation of expressed protein precursors. Azide blocks the function of SecA and the ionophors dissipate the electrochemical gradient across the cytoplasmic membrane of E. coli. Addition of azide ions resulted in the formation of inclusion bodies, highly enriched with pre-apo-plastocyanine. Plastocyanine is a soluble copper protein, which can be found in the periplasmic space of cyanobacteria as well as in the thylakoid lumen of cyanobacteria and chloroplasts, and the pre-protein contains a cleavable signal sequence at its N terminus. After purification of cyanobacterial preapo-plastocyanine, its signal sequence can be cleaved off by the E. coli signal peptidase, and protein processing was followed on Coomassie stained SDS polyacrylamide gels. We are optimistic that the presented method can be further developed and applied. PMID- 17129408 TI - 3-Hydrogenkwadaphnin induces monocytic differentiation and enhances retinoic acid mediated granulocytic differentiation in NB4 cell line. AB - Recently, we have reported that 3-hydrogenkwadaphnin (3-HK), a diterpene ester isolated from Dendrostellera lessertii (Thymealeaceae), is very effective against leukemia cell lines without any detectable effects on normal cells (Moosavi et al., 2005b). In this study, we report that 3-HK induces G1 cell-cycle arrest, differentiation and apoptosis in APL NB4 cell line. Indeed, the drug between 24 to 96 h induced 7-65% growth inhibition of NB4 cells. Cell viability was also decreased by 2-55% between 24 to 96 h treatments with the drug, respectively. These effects of the drug were also dose-dependent. According to flow cytomtry results, 3-HK (15 nM) induced a significant G1-arrest up to 24 h which was consequently followed with appearance of sub-G(1) peak at 72 to 96 h. Hoechst 33258 staining and DNA fragmentation assays confirmed the occurrence of apoptosis among the treated cells. On the other hand, NBT reducing assay, Wright-Giemsa staining, phagocytic activity and expression of cell surface markers (CD11b and CD14) confirmed that the inhibition of proliferation is associated with differentiation especially toward macrophage-like morphology. Interestingly, 3-HK at 5 and 10 nM enhanced the effects of all-trans retinoic acid (ATRA) in NB4 cells. Based on these results, 3-HK might become an ideal candidate for treatment of APL patients pending full exploration of its biological functions. PMID- 17129409 TI - Adenovirus-mediated expression of both antisense ornithine decarboxylase and s adenosylmethionine decarboxylase induces G1 arrest in HT-29 cells. AB - To evaluated the effect of recombinant adenovirus Ad-ODC-AdoMetDCas which can simultaneously express both antisense ornithine decarboxylase (ODC) and Sadenosylmethionine decarboxylase (AdoMetDC) on cell cycle distribution in colorectal cancer cell and investigated underlying regulatory responses, human colorectal cancer cells HT-29 were cultured in RPMI 1640 medium and infected with Ad-ODC-AdoMetDCas. Cell cycle progression was detected by flow cytometry analysis. The expression levels of cell cycle regulated proteins were measured by Western blot analysis. The mRNA level of cyclin D1 was measured by RT-PCR. And a luciferase reporter plasmid of cyclin D1 promoter was constructed to observe the effect of Ad-ODC-AdoMetDCas on cyclin D1 promoter activity. The results showed that recombinant adenovirus Ad-ODC-AdoMetDCas significantly induced G1 arrest, decreased levels of cyclin D1 protein and mRNA and suppressed the promoter activity. Ad-ODC-AdoMetDCas also inhibited nuclear translocation of beta-catenin. In conclusion, downregulation of ODC and AdoMetDC mediated by Ad-ODC-AdoMetDCas transfection induces G(1) arrest in HT-29 cells and the arrest was associated with suppression of cyclin D1 expression and inhibition of beta-catenin nuclear translocation. As a new anticancer reagent, the recombinant adenovirus Ad-ODC AdoMetDCas holds promising hope for the therapy of colorectal cancers. PMID- 17129410 TI - Characterization of a late gene, ORF60 from Bombyx mori nucleopolyhedrovirus. AB - Open reading frame 60 of Bombyx mori nucleopolyhedrovirus (Bm60) is located between 56,673 and 57,479 bp in the BmNPV genome which encodes 268 amino acid residues with predicted molecular weight of 31.0 kDa. Bm60 and its homologues have been identified in 11 completely sequenced lepidopteran NPVs. The transcript of Bm60 was detected by RT-PCR at 18-72 h post-infection (p.i.), while the corresponding protein could be detected at 24-72 h p.i. in BmNPV-infected BmN cells by Western blot analysis using a polyclonal antibody against Bm60. The expression of Bm60 was inhibited in the presence of Ara-c, an inhibitor of viral DNA synthesis. These results together indicated that Bm60 was a late gene. The size of Bm60 product was found to be a 31 kDa in BmNPV-infected BmN cells, consistent with predicted molecular weight. Immunofluoresence analysis showed that the Bm60 product was first detected in the cytoplasm at 24 h p.i and also located in nucleus during later infection. In conclusion, the available data suggest that Bm60 is a functional ORF of BmNPV and encodes a 31kDa protein expressed in the later stage of infection cycle. PMID- 17129411 TI - Cooperation between human DAF and CD59 in protecting cells from human complement mediated lysis. AB - The complement (C) regulatory proteins decay accelerating factor (DAF, CD55) and CD59 could protect host cells using different mechanisms from C-mediated damage at two distinct levels within the C pathway. Co-expression of DAF and CD59 would be an effective strategy to help overcome host C-induced xenograft hyperacute rejection. In this study, we made a construct of recombinant expression vector containing DAF and CD59 cDNA and the stable cell lines were obtained by G418 selection. Extraneous genes integration and co-expression were identified by PCR, RT-PCR and Western blot analysis. Human c-mediated cytolysis assays showed that NIH/3T3 cells transfected stably with pcDNA3-CD59, pcDNA3-DAF, and pcDNA3-CD59DAF DP were protected from Cmediated damage and that synchronously expressed human CD59 and DAF provided the most excellent protection for host cells as compared with either human CD59 or DAF expressed alone. Therefore, the construct represents an effective and efficacy strategy to overcome C-mediated damage in cells and, ultimately, in animals. PMID- 17129412 TI - Cloning and characterization of the HSP70 Gene, and its expression in response to diapauses and thermal stress in the onion maggot, Delia antiqua. AB - The cytosolic members of the HSP70 family of proteins play key roles in the molecular chaperone machinery of the cell. In the study we cloned and sequenced the fulllength cDNA of Delia antiqua HSP70 gene, which is 2461 bp long and encodes 643 a.a. with a calculated molecular mass of 70,787 Da. We investigated gene copies of cytosolic HSP70 members of 4 insect species with complete genome available, and found that they are quite variable with species. In order to characterize this protein we carried out an alignment and a phylogenetic analysis with 41 complete protein sequences from insects. The analysis divided the cytosolic members of the family into two classes, HSP70 and HSC70, distinguishable on the basis of 15 residues. HSP70 class members were slightly shorter in length and smaller in molecular mass relative to the HSC70 class members, and the conservative and functional regions in these sequences were documented. Mainly, we investigated the expression of Delia antiqua HSP70 gene, in response to diapauses and thermal stresses. Both summer and winter diapauses elevated HSP70 transcript levels. Cold-stress led to increased HSP70 expression levels in summer- and winter-diapausing pupae, but heat-stress elevated the levels only in the winter-diapausing pupae. In all cases, the expression levels, after being elevated, gradually decreased with time. HSP70 expression was low in non-diapausing pupae but was up-regulated following cold- and heatstresses. Heat stress gradually increased the mRNA level with time whereas cold-stress gradually decreased levels after an initial increase. PMID- 17129413 TI - Nitric oxide synthase expressions in ADR-induced cardiomyopathy in rats. AB - In this study, we investigate Nitric oxide synthase (NOS) expressions in adriamycin (ADR)-induced cadiomyopathy in rats. Sixty male Wistar rats were randomly divided into two main groups: control and ADR groups. Myocardial histopathological observation was performed; Expressions of 3 isoforms of NOS genes were examined by RT-PCR analysis; Expressions of 3 isoforms of NOS protein was assessed by Western blot analysis. Myocardium exhibited intensive morphological changes after 8 weeks of ADR treatment. The expression levels of inducible NOS (iNOS) gene and protein were significantly increased in ADRtreated rats after 8 weeks of treatment and then slightly increased at weeks 9 and 10. No significantly difference of neuronal NOS (nNOS) or endothelial NOS (eNOS) gene and protein were observed in the myocardium obtained from the control rats and ADR-injected rats at any time point. iNOS gene expression is selectively induced by ADR in heart. The upregulation of iNOS gene and protein may be somehow correlated with morphological changes seen in heart of rat treated with ADR. PMID- 17129414 TI - Fragile-X mental retardation: molecular diagnosis in Argentine patients. AB - Fragile-X-syndrome (FXS) is the most common type of inherited cognitive impairment. The underlying molecular alteration consists of a CGG-repeat amplification within the FMR-1 gene. The phenotype is only apparent once a threshold in the number of repeats has been exceeded (full mutation). The aim of this study was to characterize the FMR-1 CGG-repeat status in Argentine patients exhibiting mental retardation. A total of 330 blood samples from patients were analyzed by PCR and Southern blot analysis. Initially, DNA from 78 affected individuals were studied by PCR. Since this method is unable to detect high molecular weight alleles, however, we undertook a second approach using the Southern blotting technique to analyze the CGG repeat number and methylation status. Southern blot analysis showed an altered pattern in 14 out of 240 (6%) unrelated patients, with half of them presenting a mosaic pattern. Eight out of 17 families (47%) showed a (suggest deleting highlight). The characteristic FXS pattern was identified in 8/17 families (47%), and in 4 of these families 25% of the individuals presented with a mosaic model. The expansion from pre-mutation to full mutation was shown to occur both at the pre and post zygotic levels. The detection of FXS mutations has allowed us to offer more informed genetic counseling, prenatal diagnosis and reliable patient follow-up. PMID- 17129415 TI - Doxorubicin binds to un-phosphorylated form of hNopp140 and reduces protein kinase CK2-dependent phosphorylation of hNopp140. AB - Human nucleolar phosphoprotein p140 (hNopp140) is a nucleolar phosphoprotein that can bind to doxorubicin, an anti-cancer agent. We have examined the interaction between hNopp140 and doxorubicin as well as the folding property of hNopp140. Also, the effects of ATP and phosphorylation on the affinity of hNopp140 to doxorubicin are investigated by affinity dependent co-precipitation and surface plasmon resonance methods. Doxorubicin preferentially binds to un-phosphorylated form of hNopp140 with a KD value of 3.3 x 10(-7) M. Furthermore, doxorubicin reduces the protein kinase CK2-dependent phosphorylation of hNopp140, indicating that doxorubicin may perturb the cellular function of hNopp140 by reducing the protein kinase CK2-dependent phosphorylation of hNopp140. Low contents of the secondary structures of hNopp140 and the fast rate of proteolysis imply that hNopp140 has a high percentage of flexible regions or extended loop structures. PMID- 17129416 TI - Characterization of Choristoneura fumiferana genes of the sixth subunit of the origin recognition complex: CfORC6. AB - A new protein was cloned and identified as the sixth subunit of Choristoneura fumiferana origin recognition complex (CfORC6). The newly identified 43 kDa protein CfORC6 is much bigger than DmORC6 (25.7 kDa) and HsORC6 (28.1 kDa), though itos 23.85% identical to DmORC6 and 23.81% identical to HsORC6. Although the molecular weight of CfORC6 is close to ScORc6 (50 kDa), CfORC6 is only 14.03% identical to ScORC6. By alignment, it was found that the N-terminal of CfORC6 has about 30% identities with other ORC6s, but about 100aa of C-terminal of CfORC6 has no identity with other ORC6s. Like ScORC6, CfORC6 has many potential phosphorylation sites, (S/T)PXK. Like DmORC6, CfORC6 has leucine-rich region in the relevant site. Northern Blot showed that CfORC6 mRNA is about 2,000nt. Southern Blot confirmed that there is one copy of CfORC6 gene in spruce budworm genome. Western blot showed that infection of Cf124T cells with CfMNPV didnot affect the expression levels of CfORC6, at least up to 26 hr post infection. PMID- 17129417 TI - Identification of immunostimulatory oligodeoxynucleotide from Escherichia coli genomic DNA. AB - Bacterial DNA containing immunostimulatory CpG motifs can stimulate antigen presenting cells to express costimulatory molecules and to produce various cytokines in vivo and in vitro. In this study, we fragmented macromolecular E.coli genomic DNA with DNase I, and analyzed the ability of the resulting DNA fragments to induce the NF-kappaB activation and humoral immune response. Furthermore, using computational analysis and luciferase assay for synthetic ODNs based on the sequence of the immunostimulatory DNA fragments (DF-ODNs), an active component of DF-ODNs sequences was investigated. Experimental results demonstrated that DF-ODN is optimal for the NF-kappaB-responsive promoter activation in the mouse macrophage cell line and the humoral immune response in vivo. In agreement with the activity of the DFODNs processed by DNase I, a synthetic ODN based on the DF-ODN sequences is potent at inducing IL-12 mRNA expression in primary dendritic cells. These results suggest that the discovery and characterization of a highly active natural CpG-ODN may be achieved by the analyses of bacterial DNA fragments generated by a nuclease activity. PMID- 17129418 TI - [Problems and the considerations in endodontic treatments]. PMID- 17129419 TI - [Analysis on results of endodontic treatment and influencing factors]. AB - OBJECTIVE: To assess the clinical outcome of root canal therapy (RCT) and the various factors that may influence the outcome of RCT. METHODS: A total of 695 teeth from 357 patients were retrospectively studied three years after endodontic treatment. Pre- and intra-operative information was collected from the original patient records. The post-operative sign or symptom, periapical status and coronal restoration integrity were examined 3 years after obturation. Data were subjected to bivariate and multivariate analysis. RESULTS: The cure rate for 695 teeth was 75.1%, 96.0% of which was considered to be functional. The tooth group, pre-operative pulp and periapical status, quality of root filling and integrity of coronal restoration were revealed by means of bivariate analysis to exert a significant influence on treatment outcome. The logistic analysis indicated that the odds for cure in the teeth with pre-operative periapical radiolucency, underfilling and "open" coronal restoration were significantly lower by 2 folds, 3 folds and 1.6 folds than their counterparts, respectively. CONCLUSIONS: The pre operative periapical status, quality of root filling and the integrity of coronal restoration are main predictors of outcome in RCT. PMID- 17129420 TI - [An in vitro study on the incidence of the second mesiobuccal canal in the mesiobuccal root of the first and second maxillary molars]. AB - OBJECTIVE: To investigate the incidence of the second mesiobuccal canal (MB2) in the mesiobuccal root of the maxillary first and second molars in Chinese population using three techniques, including the clearing technique, spiral CT scanning and serial root sections. METHODS: A total of 216 extracted human first and 334 second maxillary molars were randomly divided into two groups respectively: group A and B. The teeth in group A were cleared. The specimens in group B were subjected to spiral CT scanning, and then the roots were cross sectioned every 1 mm from the root apex. Under the Dental operating microscope (DOM), the incidence of MB2 were recorded. RESULTS: (1) The incidence of MB2 in the first and the second maxillary molars were 81.48% and 49.70% respectively by clearing, and 77.78% and 47.31% from S-CT scanning, 88.89% and 53.89% respectively from serial root section. The occurrence of MB2 in maxillary first molars was statistically higher than in maxillary second molars (P < 0.05, chi square test). (2) There was no significant difference among the three approaches for detecting the MB2 canal (P > 0.05, chi square test). CONCLUSIONS: Both the maxillary first molars and the second molars have high incidence of MB2. PMID- 17129421 TI - [A short-term clinical study of one-visit endodontic treatment for infected root canals]. AB - OBJECTIVE: To clinically evaluate the postoperative pain level and short-term healing of one-visit endodontic treatment for infected root canals. METHODS: 100 infected cases were included in one-visit root canal therapy group and 42 infected cases in two-visit group using calcium hydroxide as an interappointment medicament. A comparison of the clinical results was done between the two groups. The main assessments included postoperative pain evaluated by patients and by clinical examination, and the six months' healing response. RESULTS: There was no statistically significance between the groups regarding pain 7 days after operation (one visit 60.0%, two visit 57.1%, P > 0.05) and six months' healing response (one visit 67.9%, two visit 61.5%, P > 0.05). The pain in the one-visit group was quickly relaxed postoperatively, as the pain in the two-visit group was given a rise because of the second treatment. CONCLUSIONS: One-visit endodontic treatment for infected root canals has the same clinical efficacy as two-visit in regarding to postoperative pain level and short-term healing. PMID- 17129422 TI - [Experiences of differential diagnosis for periapical diseases]. PMID- 17129423 TI - [Difficulty assessment of root canal treatment]. PMID- 17129424 TI - [Summary of the second national conference on cariology, endodontology and operative dentistry]. PMID- 17129425 TI - [Preliminary investigation of lower second molar extraction in correction of severe skeletal class III malocclusion]. AB - OBJECTIVE: To evaluate dento-skeletal and soft-tissue profile changes after extraction of lower second molars and treatment using fixed appliances in severe class III subjects. METHODS: Fourteen patients with severe skeletal class III malocclusion (male 4, female 10, age rang 12.0 - 17.1 years old, mean age 13.3 +/ 0.8), diagnosed as requiring orthognathic surgery, but rejected surgical therapy were included in the study. Lateral cephalometric films taken at the beginning and end of treatment were analyzed using Pancherz analysis and a traditional cephalometric analysis. RESULTS: After active treatment, inclination of lower incisors was decreased 11.7 degrees when measured to the mandibular plane (P < 0.001). A negative value of the distance between upper and lower lip position to SnPg' at the beginning of treatment changed to a positive value (P < 0.001). CONCLUSIONS: The results of the current preliminary study suggest that success in the treatment of some severe class III deformity in the permanent dentition can be achieved with fixed appliances and extraction of lower second molars. A remarkable soft-tissue change was noted after the treatment and concave facial profiles changed to straight profiles. PMID- 17129426 TI - [The reliability of digital periapical films in diagnosis of root resorption]. AB - OBJECTIVE: To investigate the difference between digital periapical film and conventional film in the evaluation of tooth length and root resorption. METHODS: A standard for root resorption in vitro was developed based on 20 extracted upper central incisors. Digital periapical films and conventional periapical films were taken before and after the process of simulating root resorption at six different projection angles respectively. The tooth length and root resorption were measured on these films. The results were analyzed by paired-samples rank sum test. RESULTS: Significant difference on tooth length measurement was found between two types of films on the same projection angle. No significant difference on the length of root resorption calculated on digital and conventional films was found. CONCLUSIONS: The accuracy of the digital periapical film was greater than that of conventional periapical films. The conventional periapical film can still be used in the evaluation of root resorption. PMID- 17129427 TI - [Endoscopy-assisted internal fixation of ramus and subcondylar fracture]. AB - OBJECTIVE: To investigate clinical application of endoscopy-assisted internal fixation of ramus and subcondylar fractures. METHODS: In 11 patients with mandibular ramus and subcondylar fractures, endoscopy-assisted internal fixations with titanium plates were performed using micro-angular drill and screwdriver. RESULTS: All of the patients showed no visible facial scars. Orthorpantography and three dimensional reconstructions of spiral CT scan demonstrated that all fractures were healed 1 to 15 months after operation. Slight malocclusion existed in 1 case and slight limited mouth openings were found in 2 cases. CONCLUSIONS: Endoscopy-assisted internal fixation of mandibular ramus and subcondylar fracture avoided visible facial scars and the risk of facial nerve damage, it is, therefore, a minimally invasive and effective procedure. PMID- 17129428 TI - [Segmental reconstruction of maxillary defects by arced distraction osteogenesis of zygoma]. AB - OBJECTIVE: To study the possibility of segmental maxillary regeneration by arced distraction osteogenesis of zygoma. METHODS: The distraction of zygoma with the internal arced devices was performed in 2 dogs to simulate the reconstruction of maxillary defect. The zygoma was distracted about 12 mm, and the process of new bone formation was studied by histology and scanning electron microscopy. RESULTS: The transport disk was distracted about 12 mm around the arc. After 8 weeks of consolidation, the density of new bone was close to that of normal bone. CONCLUSIONS: The segmental maxillary regeneration by arced distraction osteogenesis of zygoma is possible. PMID- 17129429 TI - [Radiation-inducible promoters-mediated cdglytk gene in the treatment of buccal carcinoma in golden hamster]. AB - OBJECTIVE: To observe the therapeutic effect of CDglyTK gene mediated by radiation-inducible promoters in the treatment of buccal carcinoma in Golden Hamster. METHODS: Animal models of buccal carcinoma in golden hamster were established by painting 0.5% dimethyl-benzanthracene. The plasmids pcDNA (+) 3.1/E-CDglyTK were transfected into tumors by lipofectamine. 24 h later, the tumors were exposed to 3 Gy irradiation. Animals were monitored at regular intervals for volume of tumors. CDglyTK mRNA was assayed by RT-PCR. Apoptosis and proliferating cell nuclear antigen were detected respectively by in situ end labeling and immunohistochemical methods. RESULTS: Compared with control groups, the tumor was suppressed obviously by CDglyTK gene therapy combined with 3 Gy induction radiation. The expression of CDglyTK gene could be detected by RT-PCR in the transfected tumor, and up-regulation of CDglyTK expression was found in tumor exposed to radiation (P < 0.05). There was significant difference in apoptosis index or proliferation index between tumor without irradiation and tumor with irradiation (P < 0.05). CONCLUSIONS: The radiation-inducible promoter can be served as a molecular switch to regulate the expression of CDglyTK gene in buccal carcinoma in golden hamster, and low dose induction radiation can significantly improve the therapeutic effects. PMID- 17129430 TI - [Relative quantification of cytokeratin 19 transcription in oral squamous cell carcinoma tissues by fluorescent quantitive real-time RT-PCR]. AB - OBJECTIVE: To investigate the relative quantification of cytokeratin 19 transcription in oral squamous cell carcinoma tissues by fluorescent quantitive real-time reverse transcription-polymerase chain reaction ((RT-PCR). METHODS: CK19mRNA level was detected by fluorescent quantitive real-time RT-PCR in cancerous and para-cancerous tissues from 31 oral squamous cell carcinoma patients. According to the 2(-DeltaDeltaCt) equation, the relative quantification fold of CK19mRNA level was calculated in cancerous tissues compared with para cancerous tissues. RESULTS: CK19mRNA levels in cancerous tissues were 2.21 folds higher than those in para-cancerous tissues, and the amplicon was specific. CK19mRNA level in cancerous tissue correlated significantly with pathological differentiation degree, the poorer the differentiation was, the higher the CK19mRNA level became. CONCLUSIONS: Fluorescent quantitive real-time RT-PCR is accurate and reliable in the detection of relative quantification of CK19 transcription in oral squamous cell carcinoma tissues. PMID- 17129431 TI - [Expression of Scleraxis in human periodontal ligament cells and gingival fibroblasts]. AB - OBJECTIVE: To study if Scleraxis, a basic helix-loop-helix type transcription factor, could be expressed in human periodontal ligament cells (hPDLC), bone marrow cells (hBMSC) and gingival fibroblasts (hGF), and to investigate if Scleraxis was involved in hPDLC differentiation. METHODS: hPDLC, hBMSC and hGF were cultured. Expression of Scleraxis in hPDLC from different passages and in hBMSC and hGF was analyzed by reverse transcription polymerase chain reaction (RT PCR). RESULTS: Scleraxis expression in hPDLC, hBMSC and hGF were significantly different (P < 0.05). The A values of Scleraxis/beta-actin in these kinds of cells were 0.877 +/- 0.024, 0.438 +/- 0.031, 0.313 +/- 0.083, respectively. The expression of Scleraxis was the highest in hPDLC and lowest in hGF. Scleraxis expression of hPDLC decreased with increase of passages in culture. CONCLUSIONS: Scleraxis was expressed in hPDLC, hBMSC and hGF in vitro, and may play an important role in differentiation of hPDLC. PMID- 17129432 TI - [Micro-tensile bond strength to sclerotic dentin in non-carious cervical lesions]. AB - OBJECTIVE: This in vitro study was to evaluate the micro-tensile bond strength (microTBS) of three adhesives to sclerotic dentin in non-carious cervical lesions. METHODS: The maxillary premolars extracted due to periodontitis and with non-carious cervical lesions were collected. The non-carious, natural cervical sclerotic lesions were bonded with a total-etching adhesive Scotchbond Multi Purpose, a two-step self-etching adhesive Contax, and an all-in-one self-etching adhesive Adper Prompt L-Pop. Artificially prepared wedge-shaped lesions were also made in sound premolars and bonded with the same adhesives as the controls. MicroTBS of these three adhesives was measured. RESULTS: MicroTBS of Scotchbond and Contax to sclerotic dentin was significantly lower than to normal dentin. But microTBS of Adper Prompt L-Pop to normal dentin was significantly lower than to sclerotic dentin. MicroTBS to sclerotic dentin was Scotchbond 46.805 MPa, Adper Prompt L-Pop 39.045 MPa, and Contax 29.852 MPa. CONCLUSIONS: In sclerotic dentin the microTBS was decreased because of the inferior micro-morphology of resin tags. Adhesives with low pH value might bond to sclerotic dentin effectively. PMID- 17129433 TI - [The application of laser holograph-speckle interferometry for measuring rotation of abutment teeth]. AB - OBJECTIVE: To explore the rotation of abutment teeth of fixed partial denture in chewing and to analyze the effect of rotation on the stability of abutment teeth. METHODS: Left mandible (without the second premolar and the first molar) was used for specimen. Holograph-speckle interferometry was employed for measuring the rotation angle. RESULTS: The maximal rotation angle was no more than 0.6 degrees and change of the angle was obviously in nonlinearity, when load increased from 9.8 N to 225.4 N. CONCLUSIONS: The laser holograph-speckle interferometry could be used to measure rotation of abutment teeth. The intricate surface contacting was the main reason for rotation and this rotation had no bad effect on abutment teeth of fixed partial denture in daily use. PMID- 17129434 TI - [Treatment of oral mucosal disease: Part II. Diagnosis and treatment of senile oral mucosal disease]. PMID- 17129436 TI - [How to write medical paper: Part IX. Requirements for English abstracts of original articles]. PMID- 17129435 TI - [Attachment denture: Part III. Clinical design for attachments]. PMID- 17129437 TI - [Chronic pain and cognitive-behavioral treatment]. PMID- 17129438 TI - [Advances of accelluar dermal matrix application in dentistry]. PMID- 17129439 TI - [Establishing the injury scale and the database of Chinese maxillofacial trauma]. PMID- 17129440 TI - [Entire restoration of deformities caused by naso-orbito-ethmoidal complex and adjacent craniomaxillofacial fracture]. AB - OBJECTIVE: To introduce the therapy of malformation caused by naso-orbito ethmoidal (NOE) complex and adjacent craniomaxillofacial fracture. METHODS: Seventy-six cases with NOE complex and adjacent craniomaxillofacial fracture underwent surgical replacement and internal fixation, using several cosmetically favorable incisions. At the same time, nasal reconstruction was performed to correct nasal deformities and defect through the coronal access during the exposure for the treatment of the NOE fracture. If larger nasal fragments were present, they were reduced and fixed by microplates or wires. If there was lack of septal support, dorsal nasal bone grafting was used to reestablish the height and anterior projection of the nose. Synthetic material (Medpor) was chosen for restoration of the orbital defects. Transnasal reduction was used for canthopexy. RESULTS: After 3 - 6 months follow-up, the outcomes of these patients were satisfactory functionally and esthetically. Posttraumatic nasal malformation and enophthalmos were corrected in most cases, and residual enophthalmos occurred in 3 cases, diplopia in 2 cases, insufficient prominence in 5 cases which underwent secondary correction with good results. Transnasal reduction of canthal realignment in the type III fracture was also satisfactory. There was no complication in this group. CONCLUSIONS: Comprehensive pre-operative evaluation of the patient and careful examinations should be taken to workout an appropriate operation plan. Simultaneous restoration for this type of complicated fracture is critical to obtain good results. PMID- 17129441 TI - [Psychological comparison of patients with soft tissue injuries in oral and maxillofacial region by different kinds of suturing]. AB - OBJECTIVE: To evaluate the psychological situations of patients with soft tissue injuries in oral and maxillofacial region by different kinds of suturing. METHODS: A total of 200 patients were selected and randomly divided into two groups. Group A received intradermic suture while group B underwent para-position suture. All patients were evaluated by hospital anxiety and depression (HAD) scales pre-suture, after one week, one month and three months. RESULTS: The HAD total scores of group B were significantly high compared with group A (P < 0.05) after one week and one month, while there was no difference between group A and group B pre-suture and three months later. CONCLUSIONS: Intradermic suture results in less psychological influence in patients with soft tissue injuries in oral and maxillofacial region. PMID- 17129442 TI - [Surgical treatment of naso-ethmoid-orbital fracture]. PMID- 17129443 TI - [Application and prospect of minimally invasive surgery in maxillofacial trauma]. PMID- 17129444 TI - [Repair of cranial and maxillofacial trauma: difficulties and solution]. PMID- 17129445 TI - [Application of the free vascularized lateral upper arm flap in intraoral reconstruction following ablative tumour surgery]. AB - OBJECTIVE: To evaluate the application of the lateral arm free flap (LAFF) in the reconstruction for intraoral defects. METHODS: Intraoral reconstruction was performed using the LAFF for 10 patients with oral squamous cell carcinoma of the tongue (n = 3), cheek (n = 4), gingiva (n = 1) and oropharynx (n = 2). There were 8 men and 2 women (mean age, 52 years). RESULTS: Nine LAFF healed uneventfully. One flap was lost on the second postoperative day due to venous insufficiency. The donor defect was closed primarily In all patients. There were no significant complications at the donor site. CONCLUSIONS: Because of the thin and pliable nature, the lateral upper arm flap can be recommended as a choice for repair of moderate-sized defects of the oral maxillofacial area. PMID- 17129446 TI - [Genetic linkage analysis and mutation detection in Chinese families with basal cell nevus syndrome]. AB - OBJECTIVE: To study the molecular genetic etiology of a Chinese pedigree with basal cell nevus syndrome. METHODS: The proband and his affected mother and a unaffected individual in the pedigree were chosen and peripheral blood was collected from them for DNA. Direct sequencing was performed to detect the mutations of PTCH gene. In order to further confirm the results of sequence analysis, all available family members were analyzed with genetic linkage analysis using 3 highly polymorphic microsatellite DNA markers in the region of 9q22.3-q31. RESULTS: No mutations of PTCH gene was detected in the proband's mother, one synonymous mutation was detected in the proband. Linkage analysis showed that the Lod scores of the 3 markers were: D9S283, Z = -2.11 (theta = 0.00); D9S1690, Z = -2.95 (theta = 0.00); D9S1677, Z = -5.94 (theta = 0.00). CONCLUSIONS: In this pedigree, mutation of PTCH gene is not related to the underlying pathogenesis of the syndrome. PMID- 17129447 TI - [Associations between periodontal status and delivery outcomes in pregnant women with a diagnosis of threatened premature labor]. AB - OBJECTIVE: To evaluate the association between periodontal conditions and delivery outcomes in pregnant women with a diagnosis of threatened premature labor (TPL). METHODS: Eighty systemically healthy pregnant women were enrolled in the study. Forty of these were pregnant women hospitalized with the diagnosis of TPL, and 40 normal pregnant women served. TPL was control clarified as TPL-PB (14 women) and TPL-TB (26 women) based on the delivery outcomes. No infants were delivered as PB in the control with non-TPL. Periodontal examinations included assessments of plaque index (PLI), clinical attachment loss (CAL), probing depth (PD), bleeding index (BI) and the percentage of periodontitis sites (PD > 3 mm, CAL >or= 2 mm). The serum level of TNF-alpha was determined using commercially available enzyme-linked immunoassays (ELISA). RESULTS: The mean PLI (0.94 +/- 0.05), percentage of periodontitis sites (2.93%) and TNF-alpha levels [14.81 ng/L (13.40 - 15.64 ng/L)] were significantly higher in the TPL group than in the non TPL group [0.59 +/- 0.03, 1.32% and 11.47 ng/L (10.82 - 12.86) ng/L] (P < 0.001). The mean PLI (0.96 +/- 0.06), BI (2.99 +/- 0.14), percentage of periodontitis sites (3.61%) and TNF-alpha levels [18.35 ng/L (15.47 - 31.94) ng/L] were significantly higher in the TPL-PB group than in the TPL-TB group [0.66 +/- 0.04, 2.76 +/- 0.12, 2.25% and 13.70 ng/L (12.64 - 14.80 ng/L)]. Significant negative correlations were observed between the gestational age at delivery and percentage of periodontitis sites as well as serum TNF-alpha levels (P < 0.05). And significant positive correlations were observed between percentage of periodontitis site and serum TNF-alpha levels (P < 0.05). CONCLUSIONS: Periodontal inflammation might be involved in the pathogenesis of preterm birth. PMID- 17129448 TI - [Novel mutations of cathepsin C gene in two Chinese patients with Papillon Lefevre syndrome]. AB - OBJECTIVE: To investigate the mutational characteristics of cathepsin C (CTSC) gene in two Chinese patients with Papillon-Lefevre syndrome (PLS), and provide molecular basis for research of the pathogenesis of PLS. METHODS: Peripheral blood samples were obtained from patients and their parents respectively. Genomic DNA were extracted after consents. Polymerase chain reaction, direct DNA sequencing and restriction enzyme reaction were performed to screen mutations of CTSC gene. RESULTS: Compound heterozygous mutations of CTSC gene were identified in the two patients. Patient I carried the G139R and S260P mutations, patient II had the R250X and C258W mutations. The parents were heterozygous carriers without the clinical feature of PLS. None of the mutations were detected in normal controls. Furthermore, the S260P and C258W changes were novel mutations of CTSC gene, which had not been reported previously. CONCLUSIONS: Mutations of CTSC gene are responsible for the phenotype of Papillon-Lefevre syndrome in two Chinese patients. The results extend the mutation spectrum of CTSC gene and also provide basis for gene diagnosis of PLS in China. PMID- 17129449 TI - [Condylar lateral movements in patients with mandibular deviation]. AB - OBJECTIVE: To record condylar movements of patients with mandibular deviation and analyze their characteristics. METHODS: Computer aided diagnosis axiography (CADIAX III) was used to record and analyze condylar lateral movements in 31 patients (9 males, 22 females and the age range 12 - 26, mean age 18 years old) with mandibular deviation. Paired-t test and rank sum test were used to compare condylar movements. Multinomial logistic analysis was used to clarify the relationship between condylar movement and mandibular deviation. RESULTS: The condylar lateral movements in patients with mandibular deviation were asymmetric. The tracing of the shifted side was longer than the contralateral side (P < 0.01). The transverse inclination of the shifted side was smaller than the contralateral side (P < 0.01). With the increase of deviation, the movement length difference between the two sides increased and the length of contralateral side decreased (P < 0.01). CONCLUSIONS: The patients with mandibular deviation not only have morphological asymmetry but asymmetric condylar movement length and inclination in lateral movements. There is specific relationship between condylar movement and mandibular deviation. PMID- 17129450 TI - [The compensation of post-treatment incisor torque in sagittal jaw relationship]. AB - OBJECTIVE: To investigate the dental compensation in different sagittal jaw relationships. METHODS: One hundred and fifty-seven post-treatment cases with satisfied outcome were chosen. Computerized cephalometric analysis was performed to evaluate the sagittal jaw relationship. Incisor torque were measured based on study models. Correlation analysis was performed. RESULTS: Compared with skeletal class I cases, the upper incisors of skeletal class III cases were more proclined, and lower incisors were more retroclined; the opposite result was found in skeletal class II cases. Dental compensation in class I cases took place primarily at lower incisors. CONCLUSIONS: Incisor torque control was important in patients with various sagittal jaw relationships. PMID- 17129451 TI - [Effects of adenovirus-mediated human bone morphogenetic protein-7 on proliferation and differentiation of human dental pulp cells]. AB - OBJECTIVE: To investigate the effect of adenovirus expressing human bone morphogenetic protein-7 (hBMP-7) on proliferation and differentiation of human dental pulp cells. METHODS: The replication-deficient adenoviral vector encoding hBMP-7 gene was constructed by using homologous recombinant modality. The efficiency of transfection was evaluated by fluorescent microscopy and flow cytometry. The expression of hBMP-7 protein in adenovirus-infected dental pulp cells was determined by Western blot. The proliferation of cells was tested by MTT method, the activity of alkaline phosphatase was assayed, von Kossa staining was used to detect mineralized nodule formation, and the expression of DSPPmRNA in cells was detected using semi-quantitative RT-PCR. RESULTS: Green fluorescent protein was visible under fluorescent microscopy. Higher transfection efficiency (91.1 +/- 1.0)% could be obtained at MOI of 75. Western blot from dental pulp cells infected with Ad-hBMP-7 for 48h detected protein expression of a hBMP-7 gene. The activity of alkaline phosphatase in cells was significantly higher than those of the control groups (P < 0.05). The cells infected with Ad-hBMP-7 had the ability of mineralization. DSPP mRNA expression of cells was in a time- and dose- dependent manner. CONCLUSIONS: Ad-hBMP-7 can induce human pulp cells into odontoblasts, but has no obvious effect on their proliferation. PMID- 17129452 TI - [Effect of chemical compounds of Galla chinensis on enamel surface rehardening in vitro]. AB - OBJECTIVE: To evaluate the effect of chemical compounds extracted from Galla chinensis on enamel surface rehardening in vitro. METHODS: Sixty bovine enamel blocks with early carious lesions were randomly divided into six groups: group1 treated NaF (positive control); group2 with GCE; group3 with GCE-B; group4 with GCE-B1; group5 with GCE-B2 and group6 with deionized water (negative control). The lesions were subjected to a pH-cycling regime for 12 days. Surface enamel microhardness was measured on the enamel blocks before and after demineralization. After pH-cycling, and the percentage of surface microhardness recovery (SMHR) was calculated. RESULTS: Obvious increase of the surface hardness of the enamel was observed in all the treatments except GCE-B2 and deionized water (P < 0.05). CONCLUSIONS: The present study demonstrates the potential of the three GCEs (GCE, GCE-B and GCE-B1) to effect net rehardening of artificial carious lesions under dynamic pH-cyclic conditions. PMID- 17129453 TI - [Quantitative study on occlusal balance of normal occlusion in intercuspal position]. AB - OBJECTIVE: To assess occlusal balance of normal occlusion in intercuspal position with maximal bite force. METHODS: Maximal bite force was recorded in intercuspal position by use of T-Scan II system from 123 subjects with normal intact dentitions. Occlusal balance of normal occlusion was quantitatively analyzed from center of force, percentage of bite force, and occlusal contacts. RESULTS: The relative position of the center of bite force, the difference in bilateral force percentage, and unsymmetrical coefficient followed normal distributions. The 95% reference ranges for corresponding testing items were -6.60 to 6.68 mm, -15.50% to 12.10%, and 0.65 to 1.39. There was no statistic difference (P = 0.915) in occlusal contacts between left and right sides. The 98.4% of normal occlusion subjects had the center of bite force locating in posterior region of dentition when biting with maximal force in intercuspal position. CONCLUSIONS: Occlusal balance could be evaluated by T-Scan II system. Occlusion of normal subjects biting with maximal force was stable and bilaterally balanced in intercuspal position. PMID- 17129454 TI - [Application of bolt-like casting post in posterior fixed partial denture]. AB - OBJECTIVE: To study the manipulation procedure and application of a new post-core system in posterior fixed partial dentures. METHODS: Abutment tooth canals were firstly prepared according to respective canal directions and then complete crown preparation was made following standard procedures. The impression of prepared abutment teeth was taken with silicone and specially made syringe. The fixed partial denture and bolt-like post were fabricated respectively and lastly the two parts were glued together in patient after try-in. RESULTS: All the fixed partial denture with the new post-core system did not get loose and there was no abutment tooth fractured after 23 months in use. CONCLUSIONS: This new post-core system could improve retention of the posterior fixed partial denture if the inter-occlusion distance of the abutment tooth was shorter than 3 mm. PMID- 17129455 TI - [Expression of Pin1, beta-catenin and cyclin D1 in salivary adenoid cystic carcinoma and its significance]. AB - OBJECTIVE: To investigate the expression of Pin1, beta-catenin and cyclin D1 in salivary adenoid cystic carcinomas (SACC) and to evaluate the role of beta catenin and Pin1 in SACC carcinogenesis. METHODS: The expressions of Pin1, beta catenin and cyclin D1 were examined in the specimens of 65 patients with SACC by immunohistochemistry, and Pin1 protein and mRNA expressions detected by Western blot and RT-PCR in four SACC cell lines. RESULTS: Pin1 was overexpressed in 51 cases of ACC (78%), and 41 (63%) cases showed positive immunoreactivity for cyclin D1 protein in the nuclear fraction in tumor tissues. Fourteen (22%) cases showed positive immunoreactivity for beta-catenin protein in the nuclear/cytoplasmic fraction in tumor tissues, 6 of which exhibited quite evident expression of beta-catenin in nucleolus. The expression of membranous beta catenin was down-regulated in most of the patients with lymph node metastasis (11/14). CONCLUSIONS: The results suggest that Pin1 and beta-catenin signalling pathway were activated in SACC and might play a pivotal role in SACC carcinogenesis and metastasis. PMID- 17129456 TI - [Expression of nuclear factor-kappaB, Ki-67 and matrix metalloproteinase-9 in calcifying odontogenic cyst]. AB - OBJECTIVE: To evaluate the expression of nuclear factor-kappaB (NF-kappaB), Ki-67 and matrix metalloproteinase-9 (MMP-9) in calcifying odontogenic cyst (COC), in order to investigate the proliferation and invasion of COC. METHODS: Twenty-six cases of COC were classified into calcifying cystic odontogenic tumor (CCOT), dentinogenic ghost cell tumor (DGCT) and ghost cell odontogenic carcinoma (GCOC) based on the WHO classification of odontogenic tumors in 2005. The specimens of COC and 10 classic ameloblastoma (AB) were examined immunohistochemically to determine the expression of NF-kappaB p65, Ki-67 and MMP-9. RESULTS: NF-kappaB was mainly detected in the cytoplasm of most tumor cells, but was only detected in the nucleus of few tumor cells (rate of nuclear staining < 1%). The expression of Ki-67 was significantly higher in GCOC than in CCOT (P < 0.001), DGCT (P < 0.05) and AB (P < 0.005). MMP-9 was detected both in tumor cells and stromal cells. GCOC showed significantly higher percentage of MMP-9 positive cases in stromal cells than CCOT, DGCT and AB (P < 0.05). CONCLUSIONS: NF-kappaB may minimally affect the progression and invasion of COC. GCOC shows significantly higher proliferative activity and aggressiveness than CCOT and DGCT. MMP-9 in stroma may play a key role in the invasion of GCOC. PMID- 17129457 TI - [Application of functionally generated path technique in computer-aided-design modeling of occlusal surface of full crown]. AB - OBJECTIVE: To apply the functionally generated path (FGP) technique for modeling the occlusal surface of a computer-aided-design (CAD) posterior full crown to obtain anatomic morphology. METHODS: A patient with defected left mandibular first molar was employed. After tooth preparation and impression making, the gypsum working cast and die were scanned with a digitized mechanical scanner and the surface data was acquired. The interocclusal records at intercuspal position (ICP) and FGP were made in the patient's mouth. These records were placed on the working cast and their surfaces were scanned. In the process of the computer aided designing full crown, the cusps and fossae of the occlusal surfaces were accurately modified according to the digitized information of ICP and FGP interocclusal records. RESULTS: A full crown was designed and the occlusal morphology of the restoration was adapted to dynamic occlusion as well as static occlusion. CONCLUSIONS: The FGP technique was practical for the CAD of full crown and could avoid potential occlusal interferences with opposing teeth during function. PMID- 17129458 TI - [Treatment of oral mucosal diseases: part III. Diagnose and treatment of syphilis and acquired immuno-deficiency syndrome]. PMID- 17129459 TI - [Attachment denture: part IV. Problems and treatment after using of attachments]. PMID- 17129460 TI - [Study of functional brain imaging in oral taste perception]. PMID- 17129461 TI - [Theophylline in the treatment of chronic obstructive pulmonary disease: a randomized, double-blind, placebo-controlled study]. AB - OBJECTIVE: To observe the benefits and safety of low-dose, slow-release oral theophylline for long-term treatment of stable chronic obstructive pulmonary disease (COPD). METHODS: This was a randomized, parallel-group, double-blind, placebo-controlled trial. Slow-release theophylline (200 mg/d) twice daily or placebo (matching theophylline) was randomly given to 110 patients with stable COPD in the rural area of Shaoguan, Guangdong Province, for one year. Efficacy measures were spirometry and exacerbations, quality of life, dyspnea scores, satisfaction with treatments and adverse effects. Comparison of benefits was performed using superiority test. RESULTS: Of 110 patients, 85 (42 subjects in theophylline group and 43 subjects in placebo group) completed the study. An analysis for intention-to-treat (ITT) individuals showed that individuals with the treatment of theophylline experienced statistically fewer numbers [(0.8 +/- 1.2) times/year, (1.7 +/- 2.6) times/year, Z = -1.674, P = 0.047] and days of exacerbations [(4.6 +/- 7.9) d, (12.5 +/- 22.8) d, Z = -1.699, P = 0.045] in comparison to subjects receiving placebo, that patients receiving theophylline were less likely than the placebo group to experience moderate exacerbations [(0.4 +/- 1.0) times/year, (1.0 +/- 1.8) times/year, Z = -2.136, P = 0.017], and that more individuals satisfied with treatments in the theophylline group than the placebo group (n = 16, 3, Z = -2.198, P = 0.014), and that statistically greater improvement in pre-bronchodilators FEV(1) [(0.006 +/- 0.180) L, (-0.053 +/- 0.169) L, t = 1.789, P = 0.038] were found in the theophylline group in comparison to the placebo group. The similar results were observed in an analysis for per-protocol (PP) subjects. Statistical improvement on quality of life was observed in the PP subjects of theophylline group than in placebo group (-28 +/- 20, -20 +/- 23, F = 2.893, P = 0.047). Time to the first exacerbation in patients receiving theophylline was also delayed in comparison to placebo (365 d, 276 d, chi(2) = 3.880, P = 0.049). But no statistical difference was found between the two groups in post-bronchodilators FEV(1) in both ITT and PP subjects (t = 0.012, P = 0.495 and t = 0.040, P = 0.484 respectively). Drug-related adverse events (8.8%) such as insomnia, palpitation, stomach discomforts or stomachache, and headache were observed in the theophylline group. CONCLUSION: Slow-released oral theophylline (200 mg/d) may be beneficial and safe in long-term treatment of stable COPD in rural area. PMID- 17129462 TI - [Long-term follow-up study of moderate chronic obstructive pulmonary disease in elderly patients]. AB - OBJECTIVE: To investigate the prognosis of moderate chronic obstructive pulmonary disease (COPD) in the elderly patients, and to evaluate the factors for long term survival. METHODS: From May 1993, a prospective cohort study was carried out, in which 191 elderly patients with moderate COPD were recruited in Shougang communities, Shijingshan District, Beijing. The general health state, arterial blood gas, pulmonary function test, electrocardiograph, chest X-ray and 6-minute walk test (6MWT) were recorded. The patients were followed for 12 years. Univariate analysis of survival rate was performed by Kaplan-Meier method. The significance test was done by Log-rank method, and the baseline factors were analyzed using the COX regression model multiplicity. RESULTS: During the 12 years of follow-up, 10 patients were lost because of migration. Twenty-six patients died of non-COPD diseases, while 52 died of COPD. The survival rate was 81.7% (156/191) in 5 years, and 56.9% (103/181) in 12 years. The result with COX multiplicity showed that the important factors for prognosis of the patients were body mass index [BMI, relative risk (RR) = 0.916, 95% confidence intervals (95% CI) = 0.841 - 0.996], forced expired volume in one second (FEV(1), RR = 0.999, 95% CI = 0.998 - 1.000), arterial partial pressure of oxygen (PaO(2), RR = 0.929, 95% CI = 0.892 - 0.967), 6MWT (RR = 0.997, 95% CI = 0.995 - 1.000) and P pulmonale of ECG (RR = 4.081, 95% CI = 1.567 - 10.624). CONCLUSION: The important factors for prognosis of moderate COPD in elderly patients included BMI, FEV(1), PaO(2), 6MWT and P pulmonale in ECG. PMID- 17129463 TI - [The possible mechanism of lung injury induced by severe acute respiratory syndrome coronavirus spike glycoprotein]. AB - OBJECTIVE: To investigate the role of severe acute respiratory syndrome coronavirus (SARS-CoV) in the induction of acute lung injury by promoting the synthesis of chemokine/cytokines in human endothelial cells. METHODS: Twenty three SARS patients were enrolled in this study, comprising 15 male and 8 female, aged 27 - 55 years, mean (36 +/- 6) years. They were treated at Guangzhou Institute of Respiratory Disease from February to May in 2003. Chemokines/cytokines in the blood of patients with SARS were dynamically screened by liquid chip system. The lung was studied histopathologically using immunohistochemical technique. Spike glycoprotein of SARS-CoV was recombined by using insect-baculovirus expression system and Nickel affinity Magnet Beads, and then used to stimulate cultured human umbilical vein endothelial cells (HUVEC). Morphological changes of HUVEC were observed by microscope. Levels of chemokines/cytokines involved in immunoreaction in response to virus infection were detected in the supernatants of those cells cultured with the Spike glycoprotein by liquid chip system. RESULTS: Interferon-gamma inducible protein 10 (IP-10) was markedly elevated in the blood during the early stage of SARS [(7,600 +/- 2,400) ng/L, P < 0.01], and remained at a high level in the progressive stage [(8,100 +/- 2,300) ng/L, P < 0.01] and the end stage [(8,000 +/ 2,800) ng/L, P < 0.01] until convalescence [(1,250 +/- 450) ng/L, P > 0.05]. Moreover, IP-10 was highly expressed in the lung. Vacuoles appeared in part of HUVEC after Spike glycoprotein stimulation. As time going on, the HUVEC turned to be round in shape and even disrupted. Under normal condition, no detectable IP-10 was found in HUVEC. A high level of IP-10 [(179 +/- 34), (889 +/- 212), (1,676 +/ 199) ng/L, all P < 0.05] was detected in the HUVEC 12 h after Spike glycoprotein (5, 20, 40 mg/L) stimulation respectively, and presented with a significant dose dependent response. CONCLUSIONS: (1) A significant increase of IP-10 activity in the blood was found in patients with SARS-CoV infection, and remained at a high level until the stage of convalescence. A strong IP-10 protein expression was also found in SARS-CoV infected lung in autopsy. (2) The Spike glycoprotein of SARS-CoV induced a high level of IP-10 in endothelial cells, which in turn damaged endothelial cells. (3) The Spike glycoprotein of SARS-CoV induced IP-10 production by a way independent of IFN-gamma. PMID- 17129464 TI - [A study on the relevance of airway inflammatory indices in induced sputum and airway hyperresponsiveness in asthmatics]. AB - OBJECTIVE: To analyze the correlations between NO(3)(-)/NO(2)(-), eosinophil counts in induced sputum and airway hyperresponsiveness (AHR) and therefore to explore the clinical significance of these parameters in severity assessment and medication adjustment in patients with mild to moderate asthma. METHODS: From February 2003 to June 2004, 35 outpatients with mild to moderate persistent asthma (mild: 9, moderate: 26) from Huaxi Hospital asthma clinic were treated with combined medications of inhaled corticosteroids (ICS) plus long-acting beta(2) agonist (LABA) for one year. The symptom scores were recorded, and AHR (represented by PC(35)sGaw), eosinophil counts and NO(3)(-)/NO(2)(-) concentrations in induced sputum were measured at regular intervals. Fifteen healthy volunteers served as control and eosinophil counts and NO(3)(-)/NO(2)(-) concentrations in induced sputum were measured. RESULTS: A total of 35 subjects were enrolled, of whom 26 completed one-year or longer follow-up. PC(35)sGaw of 26 subjects before treatment was 0.08 g/L, which became 1.40 g/L at the third months, and then maintained at a very low level (2.64 g/L) after the seventh month. NO(3)(-)/NO(2)(-) decreased from [(734 +/- 72) x 10(-3) g/L] to [(230 +/- 41) x 10(-3) g/L] by the third month (q = 6.26, P < 0.05), and [(137 +/- 27) x 10(-3) g/L] by the seventh month, which showed no significant difference with normal control (136 +/- 20) x 10(-3) g/L, q = 3.77, P > 0.05). Eosinophil counts decreased from (0.016 +/- 0.008) to (0.014 +/- 0.007) by the third month, which was not significantly different from normal control (q = 2.94, P > 0.05). In the first fifth months the concentration of NO(3)(-)/NO(2)(-) in induced sputum exhibited a significant negative correlation with PC(35)sGaw (r(1) = -0.872, r(2) = -0.653, r(3) = -0.639, r(4) = -0.656, all P < 0.05). But eosinophil counts had no correlation with PC(35)sGaw in the first three months (r(1) = 0.237, r(2) = 0.536, r(3) = 0.675, all P > 0.05). CONCLUSION: The parameters related to airway inflammation including PC(35)sGaw and sputum NO(3)(-)/NO(2)(-) may be useful in assessing asthma severity, evaluating the efficacy of treatment and adjusting medication regimens. PMID- 17129465 TI - [The relationship between tumor necrosis factor-alpha gene promoter polymorphism and obstructive sleep apnea-hypopnea syndrome]. AB - OBJECTIVE: To investigate the relationship between tumor necrosis factor-alpha (TNF-alpha) gene promoter polymorphism and obstructive sleep apnea-hypopnea syndrome (OSAHS). METHODS: The plasma TNF-alpha level of OSAHS group and non OSAHS group was detected by enzyme-linked immunosorbent assay (ELISA). Eighteen patients with severe OSAHS were treated with continuous positive airway pressure (CPAP) for 1 month, and the serum levels of TNF-alpha was also measured. The genotypes of TNF-alpha gene promoter polymorphism were determined by polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP). The genotypes and allele of the polymorphisms were compared between OSAHS group and non-OSAHS group. The effects of the polymorphisms in OSAHS group on body mass index (BMI), neck circumference (NC), waist/hip rate (WHR), polysomnography (PSG), systolic blood pressure (SBP), diastolic blood pressure (DBP) were analyzed. RESULTS: The plasma level of TNF-alpha in OSAHS group was higher than the control group [(12.3 +/- 3.62) ng/L and (8.59 +/- 1.62) ng/L, respectively, t = 7.716, P < 0.01]. CPAP significantly decreased the serum levels of TNF-alpha, but its level (10.31 +/- 1.91) ng/L was still higher in the patients than the control group. The frequencies of TNF-alpha AA/AG genotype in OSAHS group (frequencies 31/76, 40.8%) was higher than the control one (frequencies 7/42, 16.7%) (chi(2) = 7.485, P < 0.05). Statistical analysis showed that OSAHS group had a significantly higher TNF-alphaA allele frequency (frequencies 39/152, 25.7%) than that of the control one (frequencies 39/152, 9.5%) (chi(2) = 8.830, P < 0.01). The OSAHS patients with AA/AG genotype had significantly higher serum levels of TNF-alpha, NC, WHR and aphea-hypopnea index [(13.39 +/- 3.71) ng/L, (45.2 +/- 4.2) cm, (0.91 +/- 0.12), and (34.8 +/- 15.6)/h, respectively] than those with GG genotype group [(11.09 +/- 3.54) ng/L, (42.7 +/- 4.9) cm, (0.85 +/- 0.12) and (26.4 +/- 12.3)/h, respectively] (t = 2.725, 2.278, 2.150, 2.609 respectively, P < 0.05 or < 0.01). The L SaO(2) (the lowest SaO(2)) in patients with AA/AG genotype [(78.8 +/- 10.9)%] was significantly lower than that in patients with GG genotype [(83.4 +/- 8.6)%] (t = 2.039, P < 0.05). There was no significant difference in BMI, SBP and DBP. CONCLUSION: The presence of the TNF alphaA allele may be associated with susceptibility to OSAHS, and it maybe an important candidate gene for OSAHS. PMID- 17129466 TI - [Bronchial hyperreactivity in endobronchial tuberculosis]. AB - OBJECTIVE: To investigate the bronchial hyperreactivity (BHR) and pulmonary function in patients suffering from endobronchial tuberculosis (EBTB), and therefore to provide clues for early and correct diagnosis of EBTB. METHODS: Twenty-four patients with EBTB, proved by fiberoptic bronchoscopy, chest computerized tomography and biopsy, received measurements of pulmonary ventilatory function, airway resistance (Raw), and bronchial responsiveness test. The results were analyzed by t test and correlations. RESULTS: As many as 41.7% of the patients with EBTB were found to have BHR, which had never been recognized before. The patients with EBTB usually had severe cough (100%, 24/24), shortness of breath (54%, 13/24), and wheezing, but bloody sputum was found in only 21% (5/24), and so the patients tended to be misdiagnosed as having asthma, especially cough variant asthma. FEV(1%) in the group of EBTB with BHR was significantly higher than that in the group of EBTB without BHR (t = 2.345, P < 0.05). But there was no significant difference of FEV(1)/FVC%, MMEF%, V(75%) and Raw between the two groups. In the group of EBTB with BHR, FEV(1%) showed a negative correlation with BHR (r = -0.61, P < 0.05), but there was no remarkable correlation between the other pulmonary function parameters with BHR. CONCLUSION: BHR is present in a considerable number of patients with EBTB, and therefore attention should be paid to the differential diagnosis of EBTB and cough variant asthma. PMID- 17129467 TI - [The role of polymorphonuclear cells in lung ischemia-reperfusion injury in a canine model of pulmonary thromboembolism]. AB - OBJECTIVE: To explore the effects of polymorphonuclear cells (PMN) on lung ischemia-reperfusion (I/R) injury in a canine model of pulmonary thromboembolism. METHODS: Fifteen dogs were divided into three groups: a sham group (n = 5), an ischemia group (n = 5) and a reperfusion group (n = 5). PMN in the whole blood were isolated with density gradient centrifugation. Apoptosis rates of the PMN was measured through flow cytometer after the cells were labeled by Annexin V FITC before embolectomy and at 2, 4, 6 h after the operation. The myeloperoxidase (MPO) concentrations in lung homogenates were measured by ELISA. Alveolar PMN in the reperfusion lobar were observed by optical microscopy. The lung ultrastructure was studied by transmission electron microscopy. RESULTS: Alveolar PMN infiltration and the concentrations of MPO in the reperfusion group were significantly higher than those in the ischemia group (PMN (31 +/- 11) vs (8 +/- 4)/ten high power fields, MPO (11.7 +/- 1.6) vs (9.1 +/- 0.5) microg/L, P < 0.05). In the reperfusion group, abundant inflammatory cell infiltration was observed, predominantly with PMN in the alveoli. Apoptosis rates of the blood PMN at 6 h after reperfusion were much lower than before reperfusion (3.0 +/- 2.5)% vs (7.4 +/- 4.5)%, P < 0.05). At 4 and 6 h after operation, the PMN apoptosis rates in the reperfusion group were significantly lower than the ischemia group (4 h: (4.8 +/- 2.6)% vs (9.3 +/- 2.0)%, 6 h: (3.0 +/- 2.5)% vs (8.0 +/- 1.6)%, P < 0.05). PMN attaching firmly to the alveolar septum were observed by electron microscope. CONCLUSION: PMN with enhanced activities and decreased apoptosis rate, are involved in the cellular mechanisms of the lung I/R injury in this model of pulmonary thromboembolism. PMID- 17129468 TI - [Cyclin D1 and its association with airway remodeling in a murine model of asthma]. AB - OBJECTIVE: To evaluate the expression of cyclin D(1) in asthmatic mouse lungs, and to explore the role of cyclin D(1) in bronchial asthma and airway remodeling. METHODS: Forty BALB/c mice were randomized to group A (normal), group B (sensitized for 2 weeks), group C (sensitized for 4 weeks) and group D (sensitized for 8 weeks), 10 mice each group. The mice were sensitized with 10% ovalbumin and challenged with 1% ovalbumin to establish the asthmatic model. The number of eosinophils and the cell percentages in bronchoalveolar lavage fluid (BALF) were counted by cytology method. Pulmonary functions were measured to evaluate the resistance of expiration. Airway inflammation and eosinophil infiltration were evaluated by HE staining, and the airway wall thickness (WAt/Pi), smooth muscle thickness (WAm/Pi) and smooth nucleus counts (N/Pi) were quantified by computer-assisted image analysis system. The mRNA expression of cyclin D(1) was measured by RT-PCR and Real-time PCR. The protein expression of cyclin D(1) was assayed by Western blot. The correlation between airway resistance of expiration and the expression of cyclin D(1) was studied. RESULTS: The eosinophil count and differential in BALF of group B, C, and D [(42.6 +/- 0.9) x 10(4)/L, (54.7 +/- 1.4) x 10(4)/L, (44.8 +/- 2.4) x 10(4)/L] were higher than those of group A (3.4 +/- 0.5) x 10(4)/L (q = 79.75, 91.42, 84.82, all P < 0.01). The airway resistance of expiration after challenge with 45 microg/kg acetylcholine in group B, C, and D [(5.27 +/- 0.16) cmxL(-1)xmin(-1), (6.68 +/- 0.20) cmxL(-1)xmin(-1), (7.14 +/- 0.41) cmxL(-1)xmin(-1)] was higher than that in group A [(4.11 +/- 0.15) cmxL(-1)xmin(-1), q = 5.58, 6.39, 7.11, all P < 0.05]. Eosinophil infiltration, cilium loss, formation of mucus plug and smooth muscle cell layer thickening were observed in group B, C, and D. The morphological changes of the airways became evident following airway remodeling. WAm/Pi in group B, C, and D (2.8 +/- 0.6, 4.8 +/- 0.6, 6.4 +/- 0.7) were higher than in group A (2.4 +/- 0.4, q = 6.40, 8.28, 9.27, all P < 0.05), and WAt/Pi in group B, C, and D (6.4 +/- 0.8, 8.3 +/- 1.2, 9.3 +/- 1.0) were higher than in group A (5.6 +/- 1.0, q = 2.80, 4.83, 6.37, all P < 0.05). The protein expression of Cyclin D(1) in group B, C, and D (0.587 +/- 0.015, 0.808 +/- 0.029, 0.826 +/- 0.022) were higher than in group A (0.404 +/- 0.016, q = 5.87, 8.08, 8.26, all P < 0.01). There was a positive correlation between the expression of cyclin D(1) and airway resistance (r = 0.83, P < 0.05). CONCLUSIONS: The expression of cyclin D(1) in the asthmatic mouse lung was increased, and positively correlated to airway reactivity. Cyclin D(1) might be essential in the airway remodeling of asthma through ERK signaling pathway. PMID- 17129469 TI - [The effects of CpG-oligodeoxynucleotides on airway remodeling in chronic asthmatic mice]. AB - OBJECTIVE: To investigate if CpG-oligodeoxynucleotides (CpG-ODN) intervention has inhibitory effects on the development of airway remodeling in an ovalbumin (OVA) sensitized mouse model of chronic asthma. METHODS: Forty female C57BL/6 mice were randomly divided into four groups (n = 10): (1) Group A (chronic asthma model): mice were sensitized by intraperitoneal injection of OVA (10 microg) precipitated with aluminium hydroxide (100 microg) on days 1 and 14. From day 21, the mice were challenged by nebulized 2.5% OVA solution (30 min/d, three times a week for 8 weeks). (2) Group B (CpG-ODN intervention group): mice were sensitized and challenged as above, and were given 60 microg CpG-ODN by intraperitoneal injection for once every two weeks. (3) Group C (GpG-ODN control): Mice were given GpC-ODN instead of CpG motifs, other treatments same as Group B. (4) Group D (saline control): mice were sensitized and challenged by saline. All mice were killed 24 h after the final OVA challenge. Blood was obtained for eosinophil counts and measurement of serum IgE by enzyme-linked immunoabsorbent assay (ELISA). Bronchoalveolar lavage fluid (BALF) was collected for total and differential counts. The concentration of interleukin-13 (IL-13) and transforming growth factor-beta1 (TGF-beta(1)) in BALF was measured by ELISA. The left lung was isolated for pathological examination. Lung sections were stained with hematoxylin and eosin (HE), and Masson's trichrome. Other sections were prepared for immunohistochemistry using monoclonal antibodies against alpha-smooth muscle actin (alpha-SMA) and TGF-beta(1). RESULTS: The eosinophil count [(89 +/- 10) x 10(4)/ml], serum IgE [(279 +/- 53) ng/ml], BALF eosinophils [(6.30 +/- 1.30) x 10(5)/ml] and the concentrations of BALF IL-13 [(4 015 +/- 361) pg/ml] and TGF beta(1) [(356 +/- 64) pg/ml] in the OVA-sensitized mice (Group A) showed significant difference as compared with those in the NS control group (Group D, t values are 24.0, 15.7, 14.7, 18.4, 12.0 and 18.9 respectively, all P < 0.01). In Group A, the percentages of positive staining area in Masson's trichrome, alpha SMA staining and TGF-beta(1) staining were (29.7 +/- 4.2)%, (45 +/- 7)% and (34 +/- 4)% respectively. These percentages were significantly different from those in the NS control group (Group D, t values are 18.0, 15.6 and 17.9 respectively, all P < 0.01). In mice treated with CpG-ODN (Group B), the percentages of positive staining area in Masson's trichrome, alpha-SMA staining and TGF-beta(1) staining were (13.8 +/- 3.2)%, (24.7 +/- 3.1)%, (18 +/- 4)% respectively, which were significantly different from those in Group A (t values are 9.5, 8.9 and 9.8 respectively, all P < 0.05). CONCLUSIONS: This study demonstrated that CpG-ODN could prevent Th2 responses, eosinophilic inflammation and the development of airway remodeling. Its inhibitory effect on airway remodeling might, in part, be due to inhibition of the expression of cytokines such as TGF-beta(1) and IL-13. PMID- 17129470 TI - [The animal experimental study on rifampicin-dependent Mycobacterium tuberculosis]. AB - OBJECTIVE: To investigate the presence of rifampicin-dependent Mycobacterium tuberculosis strains by use of a guinea pig model of tuberculosis of rifampicin dependent Mycobacterium tuberculosis. METHODS: Guinea pigs were randomly divided into groups of infection by rifampicin-dependent Mycobacterium tuberculosis strains (1130 strain, 1219 strain, b858 strain), rifampicin-resistant Mycobacterium tuberculosis strain (1290 strain) and ATCC 35810 strain and each group was further divided into an experimental group and a control group. The guinea pigs were challenged with 1130 strain, 1219 strain, b858 strain, 1290 strain and ATCC 35810 strain to establish the tuberculosis model. The experimental groups were treated with rifampicin. The parameters including macroscopic visceral pathological change index, visceral weight index (spleen, lungs and liver), the colony-forming units (CFU) quantity of visceral Mycobacterium tuberculosis culture (spleen, lungs) and tissue pathology of guinea pigs were observed. RESULTS: At the 7th week after challenged with 1130 strain, 1219 strain, 1290 strain and b858 strain, all animals were sacrificed. The macroscopic visceral pathological change indices of the experimental group were 68.7 +/- 13.8, 60.0 +/- 13.5, 70.0 +/- 5.8 and 23.8 +/- 18.9, whereas all those parameters of the control group were 76.2 +/- 18.9, 40.0 +/- 16.8, 63.8 +/- 10.3 and 22.5 +/- 15.5 respectively, and there was no significance between the experimental group and the control group (t = 0.64, 1.85, 0.35 and 0.10, all P > 0.05). The spleen weight indices of experimental group were 0.229 +/- 0.048, 0.256 +/- 0.067, 0.324 +/- 0.054 and 0.199 +/- 0.029, whereas all those parameters of control groups were 0.278 +/- 0.025, 0.216 +/- 0.076, 0.368 +/- 0.033 and 0.213 +/- 0.038 respectively, and there was no significance between the experimental group and the control group (t = 1.75, 0.79, 1.41 and 0.57, all P > 0.05). The CFU quantity of spleen Mycobacterium tuberculosis culture of the experimental group were 4.98 +/- 0.30, 4.68 +/- 1.26, 5.07 +/- 0.47 and 3.85 +/- 0.45, whereas all those parameters of control groups were 4.90 +/- 1.03, 4.79 +/- 0.45, 5.08 +/- 0.55 and 4.23 +/- 0.95 respectively, and there was no significance between the experimental group and the control group (t = 0.11, 0.15, 0.03 and 0.73, all P > 0.05); Moreover, the tissue pathology of both groups was similar. CONCLUSIONS: The tuberculosis model of rifampicin-dependent Mycobacterium tuberculosis strains was similar to the model of rifampicin-resistant Mycobacterium tuberculosis in guinea pigs. Rifampicin-dependency was not evident in this guinea pig tuberculosis model. PMID- 17129471 TI - [The combination of recombinant rAd-p53 and adriamycin for management of primary drug resistance in chemotherapy of lung squamous cell cancer]. AB - OBJECTIVE: To investigate the combination of wild-type p53 (wtp53) gene substitution and adriamycin (ADM) on the lung cancer in vivo. METHODS: The effect of combination of recombinant Adeno-wtp53 (rAd-p53) and ADM on reversing primary drug resistance to ADM was studied for the non-small cell lung cancer (NSCLC) in a nude mice model developed by subcutaneously transplanting with the 16HBE lung cancer cell line. Twenty four nude mice with loaded tumors were randomly divided into 4 groups (no treatment, rAd-p53 treatment alone, ADM treatment alone, combination of rAd-p53 plus ADM chemotherapy). The effect of rAd-p53 substitution and ADM on the drug sensitivity was evaluated. RESULTS: In vivo studies on nude mice model transplanted with NSCLC showed that, rAd-p53 treatment alone suppressed tumor growth by 60.11% of the tumor weight and 85.4% by the volume (n = 6), while the combination of rAd-p53 with ADM suppressed tumor growth more significantly (82.32%, 99.5%, respectively, n = 6). The treatment with Adriamycin alone was less effective (35.4%, 73.9%, respectively, n = 6). CONCLUSION: Combination of rAd-p53 and ADM significantly increased the sensitivity of NSCLC tumor graft to ADM, suggesting that the combination of replication-deficient wild p53 adenovirus with DNA-damaging drugs may increase the efficacy of chemotherapy in NSCLC and overcome primary drug resistance. PMID- 17129488 TI - [Guidelines for the diagnosis and treatment of community-acquired pneumonia: learning and practicing]. PMID- 17129472 TI - [A study of the bacteriophage-based assay for the detection of pyrazinamide resistance in Mycobacterium tuberculosis]. AB - OBJECTIVE: To set up and evaluate the method of phage amplified biologically assay (PhaB) in rapid detection of pyrazinamide (PZA) resistance. METHODS: The PhaB assay was developed and applied in detecting PZA resistance in 108 clinical isolates of Mycobacterium tuberculosis and the results were compared with those of the absolute concentration method. The minimum inhibitory concentration (MIC) was detected for all discrepancy isolates. RESULTS: The results showed that the optimal detecting condition was pH 5.5, PZA 200 microg/ml and 48 h. Of the 108 strains of Mycobacterium tuberculosis, 28 strains were PZA-susceptible and 80 strains were PZA-resistant detected by PhaB; while 32 strains were PZA susceptible and 76 strains were PZA-resistant by absolute concentration method. Twenty-eight of the 108 strains were PZA-susceptible and 71 were PZA-resistant by the two methods. The concordant isolates of determination of PZA resistance were 99 by the two methods and the concordance rates was 91.7%. There were 9 strains in discordant isolates, of which 7 were the same with MIC method and gene chip in drug susceptibility. If the results of absolute concentration method was the gold standard, the sensitivity, specificity, positive and negative predictive values, as well as accuracy of PhaB assay was 94.7%, 84.8%, 93.4%, 87.5% and 91.7% respectively. CONCLUSION: The PhaB assay can be used as a rapid screening method for detection of drug susceptibility of PZA in clinical isolates of Mycobacterium tuberculosis. PMID- 17129491 TI - [Cryptogenic organizing pneumonia: CT features and the effect of glucocorticoids]. AB - OBJECTIVE: To study the CT manifestations of cryptogenic organizing pneumonia (COP) after glucocorticoid treatment. METHODS: The diagnosis of COP was made based on clinical and radiological features and confirmed by lung biopsy and pathological examination in 21 cases from 2001 to 2005. CT follow-up was carried out 5 - 75 days following therapy. The follow-up lasted 3 - 43 months. All data were analyzed and relevant literatures were reviewed. RESULTS: There were 2 male an 19 female patients with ages of 39 - 74 years. CT scans revealed multiple patchy shadows or patchy air-space consolidations, often by a predominantly subpleural and/or basal distribution. Ground-glass opacities and bronchiectasis were common findings. Migratory lesions were found in some patients. Glucocorticoid treatment resulted in significant improvement in most cases. Cure was achieved in 4 cases, significant improvement in 16, and failure in 1 case. Recurrence was found in 2 cases. CONCLUSIONS: The diagnosis of COP requires combination of clinical, CT and pathological manifestations. The effect of glucocorticoid therapy can be evaluated by repeated CT scanning. PMID- 17129492 TI - [An analysis of the clinical and radiological features of bronchioloalveolar carcinoma]. AB - OBJECTIVE: To describe the clinical and radiological features of bronchioloalveolar carcinoma. METHODS: Data of 1 050 inpatients of lung cancer, including 50 cases of pathology-proven bronchioloalveolar carcinoma, diagnosed in our hospital between 1993 and 2003, were retrospectively reviewed. The clinical information of all the patients and the HRCT findings of 31 cases of bronchioloalveolar carcinoma were analyzed. RESULTS: There was a female predominance (60%) in the 50 patients with bronchioloalveolar carcinoma. Cough was the most common presenting symptom (20/50). Twenty-four patients sought medical attention because of abnormal chest X-rays, most of which were of nodular type at the early stage. Twenty patients were completely asymptomatic at presentation. Out of the 31 cases with HRCT, eight were of the miliary type. "Bubble-like lucency" and calcifications inside nodules were present in 7 cases respectively out of the 11 cases of the nodular type. There were 9 cases each with low attenuation consolidation, pseudocavities, reticular shadowing and ground-glass opacities. The signs of "crazy paving" and "CT angiogram sign" (distinct vasculature at the background of low-attenuated consolidation at the peak of contrast scanning) were found in 2 cases. CONCLUSIONS: Bronchioloalveolar carcinoma accounted for 4.76% of lung carcinoma in our series, with a female predominance. High prevalence of asymptomatic patients at presentation and unusual long course should prompt regular chest X-ray examination. The characteristics of HRCT findings are very helpful in its diagnosis and differential diagnosis. PMID- 17129493 TI - [The value of thoracoscopy for diagnosis and management of refractory hepatic hydrothorax]. AB - OBJECTIVE: To study the value of thoracoscopy for diagnosis and management of refractory hepatic hydrothorax (HH). METHODS: Twenty-six patients with refractory HH were enrolled in this study. Twenty-four of them underwent therapeutic thoracoscopy to achieve pleurodesis by application of talc poudrage. RESULTS: Nineteen of the 26 patients had dilated chest wall veins, 6 had dilated azygos veins, and 16 had diaphragm blebs. Of the 24 patients who received pleurodesis via thoracoscope, 14 cases showed complete response, and 8 showed partial response. Mild chest pain and temperature elevation were the most complaints during or after the procedure. Liver function abnormalities were the most serious side effects after pleurodesis. During the follow-up period ranging from 6 months to 3 years, 1 patient died of upper gastrointestinal hemorrhage and encephalopathy in 1 month, 3 patients died of hemorrhage in 6, 12, 18 months respectively, and 1 case experienced recurrence in 18 months. CONCLUSIONS: Defects in the diaphragm seemed to be the main cause for the development of HH. Pleurodesis achieved by thoracoscopy and talc poudrage was effective in the treatment of HH, but complications and impaired liver functions should be considered. PMID- 17129494 TI - [Reciprocal regulation between hypoxia-inducible factor-1alpha and its prolyl hydroxylases in hypoxic pulmonary hypertension rats]. AB - OBJECTIVE: To investigate the interaction between hypoxia-inducible factors 1alpha subunit (HIF-1alpha) and its three prolyl hydroxylases (PHD1, PHD2 and PHD3) during the development of rat hypoxic pulmonary hypertension. METHODS: Forty male SD rats were randomly divided into 5 groups and exposed to normoxia (C group) or exposed to hypoxia for 3, 7, 14 or 21 d (H(3), H(7), H(14), H(21) group), respectively. Mean pulmonary arterial pressure (mPAP), vessel morphometry and right ventricle hypertrophy index (RVHI) were measured. Reverse transcriptase polymerase chain reaction (RT-PCR) and in situ hybridization were used to determine the expression of mRNA. Immunohistochemistry and Western blot were used to determine the expression of mRNA. RESULTS: The level of mPAP [(21.7 +/- 2.4) mm Hg, 1 mm Hg = 0.133 kPa], the ratio of vascular wall thickness to external diameter [WA%, (43.9 +/- 5.3)%] and pulmonary artery media thickness [PAMT, (10.0 +/- 0.7) microm] were significantly higher in H(7) group than those in C group [(16.6 +/- 1.6) mm Hg, (36.3 +/- 4.8)% and (8.5 +/- 1.3) microm respectively, q value were 5.591, 4.082, 2.929, respectively, all P < 0.05]. These parameters reached a high level and remained stable on H(14) group, and RVHI was significantly higher in H(14) group [(27.6 +/- 1.4)%] than in C group [(23.6 +/- 2.9)%, q = 5.817, P < 0.05]. HIF-1alpha protein was barely positive in C group (0.080 +/- 0.009), but markedly up-regulated in H(3) group (0.196 +/- 0.018, compared with C group q = 18.864, P < 0.05), reaching its peak in H(7) group (0.203 +/- 0.022), and then declined slightly in H(14) and H(21) group. HIF 1alpha mRNA increased marginally in H(14) group (0.176 +/- 0.019, compared with C group q = 5.401, P < 0.05, 0.139 +/- 0.017). PHD1 and PHD2 mRNA (0.260 +/- 0.031, 0.196 +/- 0.023) and protein (0.244 +/- 0.030, 0.205 +/- 0.025) were positive in C group. PHD2 mRNA and protein were up-regulated in H(3) group (0.246 +/- 0.023, 0.235 +/- 0.025, compared with C group q value was 5.268, 3.046, respectively, all P < 0.05), reaching its peak in H(14) group whereas PHD1 protein declined in H(14) group (0.210 +/- 0.023, compared with C group q = 3.885, P < 0.05) without significant mRNA change. PHD3 mRNA and protein were detected at low level in C group (0.110 +/- 0.013, 0.153 +/- 0.019), but markedly up-regulated in H(3) group (0.259 +/- 0.024, compared with C group q = 15.831, P < 0.05), and then PHD3 mRNA remained at high level while PHD3 protein declined in H(14) and H(21) group (0.206 +/- 0.025, 0.189 +/- 0.019, compared with H(7) group q value was 6.441, 8.526, respectively, all P < 0.05). Linear correlation analysis showed that HIF 1alpha mRNA and protein were positively correlated with mPAP. There was a positive correlation between HIF-1alpha protein and PHD2, PHD3 mRNA (r value was 0.580, 0.690, respectively, all P value was 0.000) but a negative correlation between HIF-1alpha protein and PHD2 protein (r = -0.704, P < 0.05). CONCLUSIONS: HIF-1alpha was regulated mainly at the protein level during the development of hypoxic pulmonary hypertension. PHD2 and PHD3 are inducible by hypoxia, possibly via elevated HIF-1alpha, suggesting that a hypoxic up-regulation of PHD acts via feedback mechanism to attenuate hypoxia induced responses. PHD may also be regulated by posttranscriptional mechanisms. PMID- 17129495 TI - [Perfluorocarbons injected intraperitoneally suppress neutrophilic infiltration in lipopolysaccharide-induced lung injury in rats]. AB - OBJECTIVE: To explore the suppressive effects of perfluorocarbon (PFC, C(8)F(18)) injected intraperitoneally on neutrophilic infiltration in lipopolysaccharide (LPS)-induced lung injury in rats. METHODS: A hundred and eight Wistar rats were randomly divided into 4 groups, and with 9 rats in each of the 3 time points respectively: Normal control group (NC group), PFC control group (PFC group), LPS induced acute lung injury group (LPS group) and PFC pre-treatment group (PFC + LPS group). The animals were injected intraperitoneally with PFC at dose of 15 ml/kg in PFC group and PFC + LPS group 48 hours before. LPS at dose of 7 mg/kg injected via penile vein induced lung injury at LPS group and PFC + LPS group and the animals were respectively killed by exsanguination at 2, 4, 6 hour points. The white blood cells and neutrophils were counted in bronchoalveolar lavage fluid (BALF) and venous blood, and the neutrophils were counted in lung tissue under microscope. The expression of E-selectin and intercellular adhesion molecule-1 (ICAM-1) were immunohistochemically detected in lung tissue as integrated optic density (IOD) measured with Image Pro Plus 5.1 software. RESULTS: (1) The counts of white blood cells of BALF [(1.98 +/- 0.21), (2.98 +/- 0.43), (3.95 +/- 0.29) x 10(6)/L at three times points respectively] and PMN percentages (0.170 +/- 0.069, 0.250 +/- 0.046, 0.351 +/- 0.054 at times points respectively) in LPS group significantly increased as compared with those in NC group [(1.27 +/- 0.20), (1.27 +/- 0.18), (1.26 +/- 0.11) x 10(6)/L and 0.041 +/- 0.008, 0.041 +/- 0.007, 0.041 +/- 0.007 at three times points respectively, t = 5.680 - 18.924, all P < 0.01]. The counts of white blood cells and PMN percentages of BALF in PFC + LPS group [(1.45 +/- 0.39), (2.67 +/- 0.44), (3.29 +/- 0.45) x 10(6)/L and 0.065 +/- 0.024, 0.102 +/- 0.033, 0.174 +/- 0.049 at times points respectively] significantly reduced as compared with those of LPS group (t = -4.224 - 12.033, P < 0.01). (2) On the contrary, the counts of white blood cells of venous blood in LPS group [(5.26 +/- 0.85), (4.38 +/- 0.39) x 10(9)/L at 4, 6 h] were significantly lower than NC group [(6.29 +/- 0.55), (6.28 +/- 0.60) x 10(9)/L, t = -3.088 and -7.946, P < 0.01], whereas the PFC pre treatment significantly increased the counts. (3) In lung tissue, the counts of PMN per field (7.56 +/- 1.81, 18.76 +/- 3.51 and 33.99 +/- 5.68), the expressions of E-selectin (IOD: 208 +/- 78, 283 +/- 67 and 625 +/- 85) and ICAM-1 (IOD: 208 +/- 78, 283 +/- 67 and 625 +/- 85) in PFC + LPS group were significantly decreased compared with LPS group (10.78 +/- 0.92, 31.55 +/- 3.00 and 54.14 +/- 5.49; 1,086 +/- 256, 1,606 +/- 408 and 3,409 +/- 1,751; 299 +/- 97, 378 +/- 67 and 817 +/- 149, respectively, t = -2.400 - 11.480, P < 0.01) at three time points. CONCLUSION: PFC pre-injected intraperitoneally significantly suppressed the neutrophilic infiltration in LPS-induced lung injury and reduced the expressions of E-selectin and ICAM-1 as the possible molecular mechanism of its effects. PMID- 17129496 TI - [Effect of 1-year specific immunotherapy with standardized house dust mite vaccine on mild to moderate allergic asthmatic patients]. AB - OBJECTIVE: To evaluate the clinical efficacy and safety of specific immunotherapy (SIT) with standardized house dust mite (HDM) vaccine on allergic asthmatic patients. METHODS: The investigation was a multicentre, randomized, double-blind, placebo-controlled clinical study. 132 patients with mild to moderate asthma who were allergic to HDM, recruited from three hospitals of China (The First Affiliated Hospital of Guangzhou Medical College, Shenyang General Military Hospital, Suzhou Children's Hospital), were randomly allocated to the active group (n = 66) or the control group (n = 66) respectively. The active group received SIT with a standardized depot Dermatophagoides pteronyssinus (Der p) extract absorbed to aluminium hydroxide (Alutard SQ, ALK-Abello, Denmark), while the control group received a placebo containing histamine dihydrochloride by subcutaneous injections for 1 year. Treatment of each group was started from the initial concentration. Updosing was performed with weekly injections from four 10 fold dose-increase vials (active group: 100, 1,000, 10,000, 100,000 SQ-U/ml, control group: 0.01, 0.1, 1.0, 10 microg/ml. 100,000 SQ-U/ml of Der p extract contains 9.8 microg/ml of the majoy allergen Der p1). The single-dose was injected weekly with 0.2, 0.4, 0.8 ml of each concentration in turn from the preceding 3 vials, totally for 9 weeks. Subsequently, the dose with 0.1, 0.2, 0.4, 0.6, 0.8 ml of the highest concentration was injected weekly from the 10th 14th week and a maintenance dose with 1 ml was reached at the 15th week. The dosing interval was then gradually increased to 2, 4, 6 weeks until the end of the first 26-weeks updosing phase (phase I, H(1)). Thereafter, the dosing continued at 6-week intervals for maintenance phase (phase II, H(2)) till the end of the complete treatment. The patient was observed for at least 30 minutes in the clinic after each injection. RESULTS: A total of 132 subjects were randomized and 129 subjects (64 in the active group, 65 in the control group) completed the whole study. Dropouts included three females because of poor compliance, being afraid of the epidemic of Severe Acute Respiratory Syndrome (SARS) and emigration respectively. Both groups were comparable at baseline in all clinical and laboratory parameters. The mean daily symptom scores in 4-weekly began to diverge at the 29th-32nd week with the active group showing a significant lower scores (0.17 +/- 0.33) than the control group (0.39 +/- 0.74, Z = 2.031, P < 0.05) till the end of the study. Significant difference was found in average daily symptom scores between H(1) (0.29 +/- 0.39) and H(2) (0.19 +/- 0.27) in the active group (Z = 2.923, P < 0.01), while no significant difference was found between H(1) (0.45 +/- 0.62) and H(2) (0.40 +/- 0.68) in the placebo group (Z = 1.885, P > 0.05). There were significant differences in subjective judgement of the improvement about overall asthmatic symptoms, exacerbation severity and exacerbation frequency between the active group (96.9%, 95.3%, 95.4%) and the control group (80.0%, 75.4%, 83.1%) with the self-evaluation questionnaire (chi(2) = 13.246, 11.576, 16.204, all P < 0.01). The trend line of daily medication scores in weekly intervals showed a significant decline. The mean daily medication score of phase II (0.20 +/- 0.36) was significant lower than that of phase I (0.33 +/- 0.67, Z = 3.344, P < 0.01), whereas the control group did not show a significant difference between phase II (0.35 +/- 0.96) and phase I (0.32 +/- 0.95, Z = 0.744, P > 0.05). After 1-year treatment, no significant differences were found in morning and evening peak expiratory flow of predicted value (PEF%), forced expiratory volume in one second of predicted value (FEV(1)/FVC), forced vital capacity of predicted value (FVC%), forced expiratory volume in one second to forced vital capacity ratio (FEV(1)/FVC), provoking dose decreasing FEV(1) by 20% (PD(20)-FEV(1)) between the active group [(97 +/- 17)%, (98 +/- 18)%, (91 +/- 11)%, (98 +/- 9)%, (82 +/- 10)%, 2.35 (7.9) micromol] and the control group [(99 +/- 19)%, (100 +/- 19)%, (90 +/- 14)%, (99 +/- 13)%, (82 +/- 9)%, 1.80 (7.8) micromol] (t = 0.170 - 0.630, Z = 0.264, all P > 0.05). Skin index (SI) to Der p in the active group (1.2 +/- 0.5) was significantly lower than that of the control group (1.5 +/- 0.6) after treatment (t = 2.395, P < 0.05). There was no significant difference in the level of serum sIgE against Der p between groups, which was 76.80 (97.0) kU/L in active group and 66.50 (99.3) kU/L in the control group (Z = 0.232, P > 0.05). The frequency of systemic adverse reactions graded by guidelines based on the position paper of European Academy of Allergology and Clinical Immunology (EAACI), was 5.7% and 1.8% of the total injections in the active group and the control group, respectively. Although significant difference was found in adverse reactions between groups (chi(2) = 4.705, P < 0.05), all were mild or moderate. The majority occurred within 30 minute after injection and were controlled well with symptomatic therapy. No any severe systemic adverse reaction was shown in the study. CONCLUSION: One year specific immunotherapy with standardized house dust mite (HDM) vaccine significantly improved symptoms and reduced medication use in mild to moderate allergic asthmatic patients. SIT also reduced skin prick test reactivity to Der p. Complying with the EAACI immunotherapy guidelines, SIT with standardized HDM vaccine was a safe treatment. PMID- 17129497 TI - [The effect of phosphoinositide-3-kinase and signal transducer and activator of transcription-6 on the proliferation of T lymphocytes in bronchial asthma]. AB - OBJECTIVE: To investigate the effect of phosphoinositide-3-kinase (PI(3)K) and signal transducer and activator of transcription-6 (STAT(6)) on the proliferation of CD(4)(+) and CD(8)(+) T lymphocytes in bronchial asthma. METHODS: CD(4)(+) and CD(8)(+) T lymphocytes of 15 asthmatic patients or 15 healthy control subjects were cultured in vitro from August of 2004 to February of 2005. The T lymphocytes of asthmatic patients were divided into four groups and stimulated with or without 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002) and IFN gamma. The four groups included a control group (group A), the LY294002 group (group B), IFN-gamma group (group C) and LY294002 + IFN-gamma group (group D). The cell cycle phases and the expression of cell cycle proteins (P27kip1, Cyclin D, Cyclin E), PI(3)K and STAT(6) were analyzed by flow cytometry. RESULTS: (1) The percentage of G(0)/G(1) phase, the expression rate of P27kip1 in CD(4)(+) and CD(8)(+) T lymphocytes from asthmatic patients were 82.0%, 44.6%, 13.2%, 4.5%; and those from the control group were 99.0%, 100.0%, 47.2%, 46.3%, respectively; the difference was significant between the two groups (Z value were 3.54, 4.23, 3.09 and 2.51, all P < 0.05). The percentage of S phase, the expression rate of Cyclin E, PI(3)K and STAT(6) in CD(4)(+) and CD(8)(+) T lymphocytes from the asthmatic patients were 18.0%, 51.7%, 7.9%, 9.3%, 7.6%, 8.7%, 8.2%, 6.3%; and those from the control group were 0.2%, 0.0%, 3.7%, 3.5%, 3.3%, 3.4%, 1.9%, 2.4%, respectively; there were significant differences between the two groups (Z value were 2.88, 4.61, 1.95, 2.06, 2.51, 2.32, 4.38 and 2.22, all P < 0.05). (2) The percentage of G(0)/G(1) phase, S phase, the expression rate of Cyclin D, Cyclin E in CD(4)(+) T lymphocytes of B group were 95.6%, 1.9%, 13.3% and 3.1%; and those of A group were 82.0%, 18.0%, 35.0%, 7.9%; there were significant differences between the two groups (Z value were 2.04, 2.23, 2.78 and 1.99, all P < 0.05). The percentage of S phase, the expression rate of Cyclin E in CD(8)(+) T lymphocytes of B group were 1.0% and 4.1%; and those of A group were 51.7% and 9.3%; there were significant differences between the two groups (Z value were 3.06 and 2.56, all P < 0.05). The percentage of G(0)/G(1) phase, S phase, the expression rate of P27kip1 in CD(4)(+) T lymphocyte of C group were 94.0%, 1.5%, 46.1%; and those of A group were 82.0%, 18.0%, 13.2%; there were significant differences between the two groups (Z value were 2.17, 2.54 and 2.81, all P < 0.05). The percentage of S phase, the expression rate of P27kip1 in CD(8)(+) T lymphocyte of C group were 10.8% and 23.1%; and those of A group were 51.7% and 4.5%; there were significant differences between the two groups (Z value were 2.67 and 2.05, all P < 0.05). The percentage of G(0)/G(1) phase, S phase, the expression rate of P27kip1, Cyclin D in CD(4)(+) T lymphocytes of D group were 97.0%, 0.0%, 40.4%, 21.5%; and those of A group were 82.0%, 18.0%, 13.2%, 35.0%; there were significant differences between the two groups (Z value were 2.73, 2.79, 2.56 and 2.10, all P < 0.05). The percentage of S phase, the expression rate of P27kip1 in CD(8)(+) T lymphocytes of D group were 92.1% and 1.7%; and those of A group were 44.6% and 51.7%; there were significant differences between the two groups (Z were 2.22 and 3.12, all P < 0.05). CONCLUSION: PI(3)K and STAT(6) may be new therapeutic targets for the treatment of asthma. Further studies are necessary to explore the relationship between PI(3)K and JAK1-STAT(6) signal pathway during the enhancement of CD(4)(+) T or CD(8)(+) T lymphocyte proliferation in bronchial asthma. PMID- 17129498 TI - [Lung transplantation for pulmonary fibrosis: report of 10 cases]. AB - OBJECTIVE: To discuss the operative technique, patient selection and perioperative management of lung transplantation for pulmonary fibrosis. METHODS: From September 2002 to December 2005, 31 cases of lung transplantations were performed in our hospital. There were 10 cases receiving single lung transplantation for pulmonary fibrosis, among them 2 received right lung transplantation with cardio pulmonary bypass, and one received contralateral lung transplantation-a left donor lung implanted in the recipient's right thorax. RESULTS: The in-hospital mortality was 10.0% (1/10). Nine patients were discharged from hospital, while one patient died of severe infection on the 137th postoperative day. During follow-up, one patient died of infection 9 months and another patient died of an accident 25 months after the operation. The 7 other patients were alive with improved lung functions. CONCLUSION: Lung transplantation is effective in improving the survival of selected patients with idiopathic pulmonary fibrosis. PMID- 17129500 TI - [Analysis of drug resistance to antituberculosis drugs of the first time retreated pulmonary tuberculosis patients in Shanghai]. AB - OBJECTIVE: To investigate antituberculosis drug resistance among the first time retreated pulmonary tuberculosis patients in Shanghai, and therefore to provide evidence for establishing retreatment regimen. METHODS: Analysis was conducted retrospectively on drug susceptibility tests to isoniazid, rifampin, streptomycin, ethambutol and para-aminosalicylates of the first time retreated pulmonary tuberculosis patients with a positive sputum culture in Shanghai Center for Disease Control from January 2002 to December 2004. RESULTS: The total drug resistance rate was 39.6%. The drug resistance rate in male and female patients was 38.5% and 44.4% respectively, the difference being not significant. The drug resistance rate in the young age group, the middle age group and the old age group were 50.0%, 41.6% and 34.6% respectively, the difference being not significant. The total drug resistance rates in the treatment failure group with standard initial chemotherapy and in the relapse group with standard regimen were both higher than in the relapse group with non-standard regimen (70.0%, 47.5%, 28.2%). So do the drug resistance rates of more than two drugs (70.0%, 18.3%, 6.4%). The multi-drug resistance rate of the failures with standard regimen was 70.0%, much higher than the relapses with standard regimen and the relapses with non-standard regimen. In relapses with non-standard initial therapy, the drug resistance rate of patients who received medications > or = 12 months was 55.0%, significantly higher than those treated for 1 - 5 months (13.0%). The drug resistance rate of patients who were treated with one to two drugs was lower than those treated with > or = 3 drugs, but no significant difference was observed (24.4% and 30.8%). CONCLUSION: Drug resistance rate varies in different types of the first time retreated pulmonary tuberculosis patients due to the history of drug use, which indicates that the current standard retreatment regimen is probably ineffective for some patients. PMID- 17129501 TI - [Analysis of ethambutol drug-resistance gene mutation in Mycobacterium tuberculosis by specific endonuclease]. AB - OBJECTIVE: Surveyor nuclease is a member of the family of plant endonucleases that cleave heteroduplexes DNA with high specificity at sites of base substitution mismatch and DNA distortion. Heteroduplex analysis by Surveyor nuclease is a relatively new method. The purpose of this study was to explore the application of this method in analysis of drug-resistance gene mutation in Mycobacterium tuberculosis. METHODS: Surveyor nuclease was used to analyze embB gene mutation of 60 Mycobacterium tuberculosis isolates, among which 45 were EMB resistant and embB gene mutation was found by sequencing, and 15 EMB-susceptible isolates without embB mutation. PCR amplification was carried out first, and then hybridized with products of H37Rv to form heteroduplex, cleaved by Surveyor nuclease, and lastly the results were shown by polyacrylamide gel electrophoresis and mutation was judged according to electrophoresis profile. RESULTS: Forty-five EMB-resistant isolates were found harboring embB gene mutation, while in the 15 EMB-susceptible isolates no mutation was found. All the 45 EMB-resistant isolates were revealed to harbor embB gene hotspot codon 306 mutation, among which 33 with ATG-->GTG, 3 with ATG-->ATT, 5 with ATG-->ATA, 2 with ATG-->ATC, 2 with ATG- >CTG. These results from Surveyor nuclease method was completely parallel to those of sequencing. CONCLUSION: Heteroduplex analysis by Surveyor nuclease may become a useful method for drug-resistance gene mutation analysis for Mycobacterium tuberculosis because of its simplicity, stability, low-cost, and high sensitivity and specificity. PMID- 17129502 TI - [Phrenic nerve conduction of obstructive sleep apnea-hypopnea syndrome patients]. AB - OBJECTIVE: To investigate the significance of phrenic nerve conduction time (PNCT) and diaphragm compound muscle action potential (CMAP) detection in the assessment of severity and response to treatment in obstructive sleep apnea hypopnea syndrome (OSAHS) patients. METHODS: Sixteen healthy volunteers (control group), 8 primary snorers (snorer group), 14 mild-moderate OSAHS patients (mild moderate group) and 18 severe OSAHS patients (severe group) were recruited for the study from January to July in 2005. Multi-pair esophageal electrodes were used to measure PNCT and CMAP of diaphragm in response to unilateral magnetic stimulation. The study was repeated in 5 OSAHS patients after effective nCPAP treatment for at least 2 months. RESULTS: The PNCT in severe OSAHS group was (8.9 +/- 1.2), (7.9 +/- 1.5) ms respectively, which was significantly longer than those in the control group (6.5 +/- 0.7), (6.0 +/- 0.5) ms, snorer group (6.5 +/- 1.2), (6.0 +/- 0.8) ms and mild-moderate group (7.3 +/- 1.0), (6.3 +/- 0.7) ms. The amplitude of diaphragm CMAP was (1.4 +/- 0.4), (1.4 +/- 0.3) mV in mild moderate group and (0.9 +/- 0.4), (1.1 +/- 0.6) mV in severe group, which was significantly lower than those in the control group (2.3 +/- 0.9), (2.1 +/- 0.9) mV and snorer group (1.9 +/- 0.5), (2.1 +/- 0.7) mV, and severe patients have significantly lower CMAP than mild-moderate patients. The PNCT and CMAP of both sides for all subjects correlated significantly with oxygen desaturation index and apnea-hypopnea index. The PNCT shortened significantly after effective nCPAP treatment, which was (8.6 +/- 0.6) ms, (7.4 +/- 0.5) ms for left side and (7.8 +/ 0.6) ms, (6.5 +/- 0.5) ms for right side. CONCLUSION: PNCT and CMAP detection with multi-pair esophageal electrodes in response to unilateral magnetic stimulation may be useful for the severity assessment and evaluation of response to effective treatment in OSAHS patients. PMID- 17129508 TI - Neurohormonal modulation in heart failure of ischemic etiology: correlates with left ventricular remodeling. AB - The natural progression of heart failure (HF) is accompanied by the compensatory activation of cardiac and extracardiac neurohormonal systems and changes in the anatomy and function of the left ventricle. The processes of ventricular remodeling and neurohormonal activation are mutually influenced so that strong associations may be observed between circulating concentrations of biomarkers (especially the natriuretic peptides originating from the heart) and parameters of left ventricle size and function. Temporal changes in the concentration of natriuretic peptides convey incremental prognostic value compared with a single determination and are usually related to concomitant changes in markers of left ventricle remodeling in patients with chronic HF. Whether knowledge of biomarkers' concentration can improve the clinical outcome of patients with HF by helping in guiding and monitoring their therapy treatment is a stimulating working hypothesis still to be verified. PMID- 17129509 TI - Diabetes and heart failure in the post-myocardial infarction patient. AB - Diabetes mellitus, a disease which is increasing in prevalence, is a major risk factor for coronary heart disease. In patients following acute myocardial infarction, the presence of diabetes is a powerful risk factor for the development of heart failure, and this intersection of heart failure and diabetes following myocardial infarction carries substantial risk. The poor prognosis associated with heart failure in diabetic patients following myocardial infarction is likely multifactorial. Aggressive strategies for prevention and treatment of heart failure are crucial to reducing the risk associated with diabetes and heart failure following myocardial infarction. This review summarizes epidemiologic, pathophysiologic, and therapeutic data related to diabetes and heart failure in the post-myocardial infarction setting. PMID- 17129510 TI - Left ventricular diastolic function following myocardial infarction. AB - An acute myocardial infarction causes a loss of contractile fibers which reduces systolic function. Parallel to the effect on systolic function, a myocardial infarction also impacts diastolic function, but this relationship is not as well understood. The two physiologic phases of diastole, active relaxation and passive filling, are both influenced by myocardial ischemia and infarction. Active relaxation is delayed following a myocardial infarction, whereas left ventricular stiffness changes depending on the extent of infarction and remodeling. Interstitial edema and fibrosis cause an increase in wall stiffness which is counteracted by dilation. The effect on diastolic function is correlated to an increased incidence of adverse outcomes. Moreover, patients with comorbid conditions that are associated with worse diastolic function tend to have more adverse outcomes after infarction. There are currently no treatments aimed specifically at treating diastolic dysfunction following a myocardial infarction, but several new drugs, including aldosterone antagonists, may offer promise. PMID- 17129511 TI - Epidemiology of heart failure and left ventricular dysfunction after acute myocardial infarction. AB - The development of heart failure and/or left ventricular systolic dysfunction has long been regarded as an ominous complication, significantly increasing the morbidity and short- and long-term mortality of survivors of acute myocardial infarction. Although the incidence of heart failure after myocardial infarction has fallen over the last few decades, it remains common, complicating up to 45% of infarcts. Moreover, up to 60% of myocardial infarcts will result in left ventricular systolic dysfunction, depending on the exact definition used. Those at greatest risk of developing heart failure are the elderly, females, and those with prior myocardial infarction. Advances in the management of acute myocardial infarction have led to reduced in-hospital mortality (even when complicated by heart failure), but longer-term mortality remains high in these patients. PMID- 17129512 TI - The link between obstructive sleep apnea and heart failure: underappreciated opportunity for treatment. AB - Obstructive sleep apnea (OSA) is a newly recognized risk factor for the development of systemic hypertension, ischemic heart disease, and congestive heart failure. Mechanisms responsible for these links include OSA-related hypoxemia and arousal from sleep-induced increased sympathetic activity, large negative intrathoracic pressure-induced increased left ventricular transmural pressure gradient and impaired vagal activity, plus formation of oxygen radicals. Secondary phenomena include increased platelet aggregability, insulin resistance, and endothelial dysfunction with reduced endogenous nitric oxide production. Safe, nonpharmacologic, nonsurgical therapy, namely continuous positive airway pressure, can attenuate OSA and improve cardiac function and quality of life. Searching for signs or symptoms of OSA from the patient (or bed partner), namely loud habitual snoring, apneas, nocturnal choking, orthopnea, paroxysmal nocturnal dyspnea, excessive daytime sleepiness, or cardiovascular disease which is difficult to control, may reward the curious physician with another treatment avenue. PMID- 17129513 TI - Indications for cardiac defibrillators in patients with congestive heart failure. AB - Sudden cardiac death (SCD) is a significant cause of mortality in patients suffering from heart failure and left ventricular dysfunction. Implantable cardiac defibrillators have been shown to effectively reduce the incidence of SCD in this population. Multiple clinical trials have redefined the indications and patient profiles for their use: from secondary prevention to primary prevention of SCD. In this article, we will review the clinical trials contributing to the current practice guidelines, which include device therapy. PMID- 17129514 TI - [Trends of hospitalizations for chronic heart failure in Andalusia in the last decade]. AB - BACKGROUND AND OBJECTIVES: The cost of hospitalization represents the greatest proportion of total expenditure due to heart failure. Our objective was to analyze the trends of morbidity of chronic heart failure in Andalusia between 1990-2000. MATERIAL AND METHODS: The data on hospitalizations in Andalusia (title 428 of the ninth revision of the International Disease Classification) were obtained from the National Survey of Hospital Morbidity of the National Institute of Statistics. The rates, standardized by age and gender, of admission due to heart failure were calculated by the direct standardization method. RESULTS: The absolute number of hospitalizations due to hear failure in people over 45 years was 4,345 in 1990 and 10,153 in 2000 (a relative increase of 230%) and it represents 14.2% hospitalizations in Spain. The increase was focused on those over 65 years and the standardized rates were slightly greater in women than in men. CONCLUSIONS: Hospitalization discharge rates increased mostly in the population older than 65 and women showed hospitalization rates slightly greater than men. PMID- 17129515 TI - [High cardiovascular risk due to inadequate control of risk factors in Spanish hypertensive patients seen in the Spanish primary care]. AB - INTRODUCTION: Cardiovascular diseases are the main cause of morbidity-mortality in Spain. Hypertension and the other cardiovascular factors are important in the pathophysiological basis of this fact. Control of the main risk factors in treated hypertensives was evaluated in this study. PATIENTS AND METHODS: Multicenter study in hypertensive patients treated in Primary Care. Blood pressure, cardiovascular risk factors as well as registered target organ damage and clinical diseases were measured. Finally, global cardiovascular risk was calculated. RESULTS: Only 17.3% of pharmacologically treated hypertensives were controlled (3.9% of diabetic patients). LDL-cholesterol control deteriorated as the patient's cardiovascular risk pattern increased (90.4% in low risk and 19.9% in high-very high risk). High levels of serum creatinine were detected in 8.3%, microalbuminuria being positive in 15.6%, with 20.8% ventricular hypertrophy. A total of 31% of the patients were diabetic, 59.8% of them having HbA1c > or = 6.5%. Global CV risk was high very high in many subjects (65.4%) when European Society of Hypertension-European Society of Cardiology (ESH-ESC) guidelines were used. This percentage was 3.6% when the Spanish-validated Framingham score was used. CONCLUSIONS: Blood pressure control as well as main cardiovascular risk factors is poor in this population. Considering this fact plus the number of target organ damage, the global risk is high in many patients according to the European guidelines. Guidelines that do not consider target organ damage do not estimate adequately the risk of this uncontrolled population. This conclusion could explain the clinical tendency of many doctors when attending uncontrolled patients. PMID- 17129516 TI - [Thrombolytic treatment in acute ischemic stroke in a center without previous experience. Development of internal organization and first results]. AB - BACKGROUND AND OBJECTIVE: Treatment of acute ischemic stroke within three hours with intravenous tissue-type plasminogen activator (t-PA) has been recently approved by the European Drug Agency. We present the development of an internal organization system that has permitted thrombolytic treatment in our center without previous experience as well as the results of the first year. PATIENTS AND METHOD: Development of the thrombolysis educational program for the staff informed, of the internal organization system, and combined care protocols among the participating services. Prospective registry of patients treated with t-PA within the period 1/2004-2/2006. We collected demographic data, stroke assessment scales score (NIHSS), time to treatment, seven day and three months mortality, symptomatic hemorrhagic transformation, systemic bleedings, functional independency at three months, early significant improvement and significant deterioration. RESULTS: Fifty-three patients were treated. Mean age: 65 +/- 13 years; 56% women. Mean NIHSS pre-treatment: 14 +/- 4.7. Mean time to hospital arrival: 62 +/- 40 minutes; door-to-treatment: 68 +/- 22 minutes, and mean time from stroke onset to treatment: 130 +/- 31 minutes. Symptomatic hemorrhagic transformation: 5.8%. Systemic bleeding: 3.8%. Seven day mortality: 5.6%; three months mortality: 15.1%. Early significant improvement: 51%. Significant neurological deterioration: 7.5%. Functional independency at three months: 51%. CONCLUSIONS: Treatment of acute ischemic stroke within three hours with intravenous t-PA is safe and is associated with a favourable outcome when it is applied by neurologists specifically trained in acute stroke management. PMID- 17129517 TI - [Intravenous immunoglobulin therapy: ELISA measurement of antidesmogleins 1 and 3 in three patients with pemphigus vulgaris]. AB - Intravenous immunoglobulin therapy (IVIg) has been used in the treatment of autoimmune bullous diseases unresponsive to conventional therapy in recent years. The action mechanism, which is not well known, suggests a wide spectrum of immunoregulation. In the last five years, several studies on patients with unresponsive pemphigus vulgaris with a clinical and serological outcome after IVIg administration in 80%-90% of cases have been published. We report the case of 3 patients with pemphigus vulgaris in whom we measured autoantibody titers to desmoglein 3 and 1 during 8 months. In spite of the clinical improvement, no significant decrease in antibody concentration was observed. Therapy with IVIg, although it has clinical benefit, did not decrease antibody values in our patients and thus it may need to be combined with immunosuppressant that inhibit pathogen antibody production. PMID- 17129518 TI - ["Natural" treatments of hypercholesterolemia]. AB - In this article we briefly review the evidence on the effect of different "natural" products on cholesterolemia. Plant stanols and sterols reduce cholesterol intestinal absorption and decrease total and LDL cholesterol by approximately 10%. Polycosanol is a mixture of saturated alcohols that seem to inhibit cholesterol hepatic synthesis and decrease total and LDL cholesterol by up to 25%. The effects on the cholestorolemia of soy and soluble fiber are modest. PMID- 17129519 TI - [Virus and systemic vasculitis]. AB - The participation of genetic and environmental factors has always been invoked in the pathogenesis of the autoimmune systemic diseases, including the primary vasculitides. Among the environmental factors, infections, fundamentally those having a viral nature, have always been focused on, especially after the discovery of the close existing relationship between the polyarteritis nodosa and the hepatitis B virus, on the one hand, and mixed cryoglobulinemia and the hepatitis C virus, on the other. The present review summarizes data from the most recent literature related to associations between virus infections and primary vasculitides, following the Chapel-Hill vasculitis classification. PMID- 17129520 TI - [Strategies for effective control of arterial hypertension in Spain. Consensus document]. AB - Blood pressure (BP) control is inadequate among treated hypertensive patients in Spain. Control rates are lower than 40% of all treated patients and the cause of this problem is multifactorial. Despite the fact that possible solutions to this problem have been repeatedly suggested by expert groups along the last 10 years, BP control rates are still low. This fact have a negative impact on cardiovascular morbidity and mortality of patients with hypertension. The aim of the present document has been to achieve a consensus on effective specific measures in order to improve hypertension control rates in Spain. These measures involve health care professionals (physicians, nurses, pharmaceutics), health care authorities and patients. The document summarizes the consensus conference of several scientific societies involved in cardiovascular medicine in five group of measures: a) improvement of the methodology of office BP measurement; b) improvement of compliance to treatment by patients; c) clarification of pressure targets to be achieved in hypertensive patients; d) optimization of life style modifications and pharmacological treatment of hypertension; and e) continuous medical education. The document emphasize life style changes as a crucial aspect to be implemented in all patients. These changes have a beneficial impact on pressure reduction, contribute to a better control of associated cardiovascular risk factors, and increase the effectiveness of antihypertensive drugs. Health care professionals should base their clinical practice on the recommendations of guidelines in order to modify their therapeutic attitudes in patients whose targets have not been achieved. PMID- 17129521 TI - [Skin lesions and acute renal failure after percutaneous transluminal angioplasty]. PMID- 17129522 TI - [Non-specific consciousness during the last six months]. PMID- 17129523 TI - [Respiratory insufficiency, vomiting and diarrhea in a Down's syndrome patient]. PMID- 17129524 TI - [76 year old male with polymyalgia and halo sign]. PMID- 17129525 TI - [82 year old male with stupor]. PMID- 17129526 TI - [Fever and pectoralis major muscle lesion]. PMID- 17129527 TI - [62 year old woman with orogenital apththosis, eye lesions and chondritis]. PMID- 17129528 TI - [Case report: a 47 year old male with cervical pain]. PMID- 17129530 TI - [Geriatrics and patients with multiple diseases]. PMID- 17129531 TI - [Diabetic neuroarthropathy]. PMID- 17129532 TI - [A syndrome of yellow nails: a case associated with Mycobacterium tuberculosis]. PMID- 17129533 TI - [Epidemiology, initial management and analysis of morbidity-mortality of severe burn patient]. AB - OBJECTIVE: Describe the epidemiological characteristics of severe burn patients and analyze the factors related with morbidity-mortality. DESIGN AND SCOPE: Observational, retrospective study of patients admitted to an intensive care unit of a level III hospital due to severe burns from January 1998 to December 2004. PATIENTS: 59 patients with criteria of "severe burn" and expected stay in ICU greater than three days. MAIN ENDPOINTS OF INTEREST: We studied epidemiological endpoints of this type of patients, diagnosis and initial treatment, early complications and morbidity-mortality. RESULTS: The burned body surface was 41% +/- 25% and age 49 +/- 21 years. Patients remained hospitalized in ICU for a median of 4 days (interquartile range: 2-19). A total of 78% of the patients needed mechanical ventilation, 47% had some infection during admission and 28% developed acute kidney failure during the first week. Mortality in the ICU was 42%. Endpoints associated independently with a significant increase of mortality were burned body surface greater than 35% (OR 1.08; 95% CI: 1.03-1.12) and development of kidney failure (OR 5.47; 95% CI: 2.02 -8.93). CONCLUSIONS: Mortality of these patients is very high and is conditioned largely by initial care. Percentage of burned body surface (BBS) and kidney failure entails greater mortality in our series. PMID- 17129534 TI - [Sedoanalgesia with remifentanil in definitive pacemaker implant]. AB - OBJECTIVE: Describe the use of remifentanil in definitive pacemaker implant. DESIGN: Prospective, observational study. SCOPE: Intensive Care Unit of two general hospitals. PATIENTS: Ninety-four patients subjected to DPM implant under sedation with remifentanil. INTERVENTIONS: The protocol for DPM implant was conducted: premedication with metoclopramide, remifentanil perfusion (20 micro g/ml), local infiltration with mepivacaine 2%, administration of magnesium metamizole at the end of the implant and posterior discontinuation of remifentanil. Remifentanil perfusion was initiated at 2 micro g/minute, increasing it until reaching a sedation grade 2-3 on the Ramsay scale, with a maximum of 6 micro g/minute. MAIN ENDPOINTS: Time needed to reach the desired sedation grade and duration of sedation, maximum dose of remifentanil necessary, frequency that another sedation was needed and of adverse events were recorded. Continuous quantitative endpoints were expressed as mean +/- SD. RESULTS: A sedation grade 2-3 was achieved with a perfusion rhythm of 3.6 +/- 1.4 micro g/min, in 20 +/- 22 minutes. In 89 patients (94.6%), the implant was performed only with remifentanil. Frequency of adverse events were nauseas/vomiting 21.3%, hypotension 5.3% and respiratory depression 1%. Remifentanil perfusion was discontinued in 3 patients (3.2%) due to appearance of adverse events. Another sedoanalgesic was used in 2 patients (2.1%). CONCLUSIONS: Remifentanil is useful in the implant of DPM as a sedoanalgesia method. Serious undesired effects are rare. Future studies are necessary to completely establish its effectiveness and safety in these types of procedures. PMID- 17129535 TI - [Pathophysiology of gas exchange in ARDS]. AB - ARDS is produced in a pulmonary edema picture due to increased vascular patency. In this way, the initial alteration consists in an alveolar occupation due to protein rich edema. This occupation reduces the alveolar surface available for gas exchange, increasing the pulmonary areas with poor or null V/Q ratio. As ARDS progresses, vascular phenomena occur that affect the gas exchange differently, giving rise to heterogeneity in the V/Q ratio. This situation worsens due to the appearance of areas with null ventilation in relationship with the appearance of atelectasis in lung dependent zones. All these factors form the hypoxemia picture refractory to the increase of the inspired oxygen fraction characteristic of this clinical entity. In this article, we make a review of these physiological mechanisms and the effect on the oxygenation of different ventilatory and drug maneuvers. PMID- 17129536 TI - [Monitoring pain: recommendations of the Analgesia and Sedation Work Group of SEMICYUC]. AB - In critically ill patients, pain is frequently underestimated and so insufficiently managed. Psychological, haemodynamic and neuroendocrine responses, secondary to untreated pain, could produce morbidity and even increases in patient mortality. All members of the intensive care team must have abilities to assess and to manage pain. The evaluation of pain in the critically ill patient is very difficult but extremely important. Self-reported pain is the starting point for treatment. The pain scores recommended are, VAS (visual analogue scale) and NRS (numeric rating scale) in communicative patients and Campbell scale in uncommunicative patients. Adequate and regular patient assessment leads to improved pain control. Scores higher than 3 points should not be permitted. A pain-free Intensive Care Unit should be a quality standard healthcare aim. PMID- 17129537 TI - [Invasive pulmonary aspergillosis in the non-neutropenic critical patient: future challenges]. PMID- 17129538 TI - [Sleep quality in ventilated patients: is the ventilatory method important or its adjustment?]. PMID- 17129539 TI - [Utility of vasopressin in cardiopulmonary resuscitation]. AB - Many vasopressants have been studied in cardiopulmonary resuscitation (CPR) to increase cerebral and coronary perfusion. Although there is a debate on the utility of epinephrine, this is the one that has been used historically, above all after verifying that other agents such as norepinephrine, metoxamine or phenylephrine, have not been shown to be more effective. Currently, due to the good experimental results, the use of vasopressin (ADH) in CPR is being evaluated. However there is little (only three studies) and debated evidence based on randomized clinical trials (norepinephrine or ADH) in humans. Once these are reviewed, it can be concluded: The results of the three randomized studies in humans obtain different results regarding the utility of ADH in cardiorespiratory arrest (CRA) secondary to ventricular fibrillation, electro-mechanical dissociation or asystole. More prospective studies are needed to know the role of ADH in prolonged CRA and in asystole, that may be the subgroups that can benefit the most from this drug. The neurological repercussion of a drug in the context of CRA should be evaluated before its inclusion in the CPR guides. PMID- 17129540 TI - [MARS (Molecular Absorbent Recirculating System) as hepatic extracorporeal care in serious acute liver failure of alcoholic etiology]. AB - Acute liver failure with criteria of seriousness and alcoholic etiology is a picture of worldwide distribution, with an elevated mortality, of approximately 50%, that requires admission to the ICU. Acute serious alcoholic hepatitis is defined by a Maddrey score value > 32 and/or spontaneous encephalopathy. Together with the classical treatment with corticosteroids and nutritional supplements, alcoholic abstinence and others such as anti-TNF therapy, we introduce MARS (Molecular Adsorbent recirculating System) as a extracorporeal liver assistance system with detoxification function, that is presented as a support measure that makes it possible to maintain the patient in good conditions until an organ becomes available or until the functional recovery of the native liver. In our case, MARS has shown some spectacular results and above all, results maintained over time, associated to the rest of the therapeutic measures characteristic of this disease. PMID- 17129541 TI - [Spontaneous gaseous gangrene of the pancreas and pneumoperitoneum without visceral perforation due to Clostridium perfringens]. PMID- 17129542 TI - [Acute pancreatitis, acute kidney failure, metformin intoxication and villous rectal adenoma]. PMID- 17129543 TI - [Meningococcal sepsis associated to ST segment and cardiac enzymes elevation]. PMID- 17129544 TI - [Images in Intensive Medicine]. PMID- 17129545 TI - [Comparison of esophageal stenoses produced by endoscopic sclerotherapy versus variceal ligation]. AB - INTRODUCTION: Variceal ligation (VL) eradicates esophageal varices faster than endoscopic sclerotherapy (ES) with a lower rebleeding rate and fewer secondary effects. However, most studies have evaluated the short-term effects of these treatments and some late complications may be overlooked. PATIENTS AND METHODS: To determine the incidence and the characteristics of stenosis, we included 253 cirrhotic patients treated endoscopically for variceal bleeding from 1988 to 2004 in our hospital. ES was carried out with ethanolamine 5% and polidocanol 1.5%. ES and VL were carried out every 15 days until varices were eradicated and then at 3 , 6- and 12-month intervals; if varices reappeared, the initial treatment was repeated. Stenosis was considered mild when esophageal size was more than 10 mm and severe when the endoscope could not be passed through the stricture. RESULTS: We found stenosis in seven out of 105 (6.7%) ES-treated patients and in 10 out of 148 (6.7%) VL-treated patients. The clinical characteristics of the patients and the previous number of endoscopic sessions were similar in both groups. Four out of seven ES patients developed stenosis during the first eradication process (mean: 11 months, 1-60), but this early stenosis was observed in one out of 10 VL patients (mean: 20 months, 1-72). Stenosis was severe in three out of seven ES patients (43%) but in only two out of ten VL patients (20%) (NS). CONCLUSIONS: The incidence of esophageal stenosis was similar after treatment of esophageal varices with ES and VL, although VL had a tendency to produce later stenosis. PMID- 17129546 TI - [Advances in radiography of the small intestine: computed tomography enteroclysis]. AB - Computed tomography (CT) enteroclysis is a new technique consisting of helical CT of the abdomen and pelvis after administration of water through a nasojejunal tube and intravenous contrast, resulting in adequate distension and visualization of the small bowel wall. The use of this technique is especially recommended in patients with gastrointestinal bleeding of unknown etiology, possible neoplastic processes of the small bowel, partial small bowel obstruction, and inflammatory bowel disease. One-hundred consecutive patients underwent CT enteroclysis (multiple detectors; 8, 16, or 64) over a 1-year period for suspected lesions of the small bowel. Of these, 31 were positive: Crohn's disease (17), tumors (8), partial obstruction (2), radiation enteritis (1), sprue (1), pneumatosis cystoids (1), and dilatation of bowel loops (1). In 28 of the 31 patients, the findings were confirmed by pathology, endoscopy or clinical follow-up. PMID- 17129547 TI - [Sequential combination therapy with prednisone, lamivudine and interferon alfa 2b for HBeAg-positive chronic hepatitis B]. AB - INTRODUCTION: Despite the development of potent, new antiviral drugs, the percentage of HBeAg seroconversion is approximately 35%. Immunosuppression before antiviral administration has recently been investigated with contradictory results. We evaluated the safety and efficacy of lamivudine and interferon alfa 2b with prior immunosuppression with prednisone in patients with HBeAg-positive chronic hepatitis B (CHB). METHODS: A randomized controlled study was conducted in a sample of 44 patients with HBeAg-positive CHB and persistently elevated alanine transferase (ALT) levels. The patients were distributed into two groups: 22 patients received prednisone 40 mg daily for 4 weeks, followed by 2 weeks without treatment, and lamivudine 150 mg daily for 4 weeks; lamivudine plus interferon alfa 2b (10 MIU every other day) was then administered for 24 weeks followed by continuous lamivudine 150 mg daily to complete 58 weeks. A further 22 patients received the same treatment regimen and duration, but without prednisone. RESULTS: Virologic response defined as HBeAg seroconversion plus a decrease of serum HBV DNA < 105 copies/ml 24 weeks after concluding the treatment was observed in 68% of the patients receiving previous immunosuppression compared with 54% of the control group (p = 0.26). Forty-five percent of patients with prednisone priming showed histologic improvement compared with 23% of the control group (p = 0.10). A significant proportion of patient with previous immunosuppression showed improvement in necroinflammatory activity (45% vs 23%) and fibrosis (50 vs 23%) compared with the control group. CONCLUSIONS: Virologic response was clinically, but not statistically, superior in the group with prednisone priming. Histologic improvement was notable in the group with previous immunosuppression. PMID- 17129548 TI - [Fulminant liver failure associated with T-cell non-Hodgkin's lymphoma and hepatitis C virus: a case report]. AB - Hematological malignancies can affect the liver, without producing severe hepatic involvement. We report the case of a 57-year-old man with hepatitis C virus infection and mild chronic hepatitis without antiviral treatment, who developed an aggressive T-cell non-Hodgkin's lymphoma confirmed by histological studies including liver, lymph nodes and bone marrow. The patient developed massive hepatic infiltration and acute liver failure. Rescue chemotherapy was administered but the patient died soon after with severe lactic acidosis. The immunopathological features of this association and the few reports of cases presenting with acute liver failure are reviewed. PMID- 17129549 TI - [Intestinal leishmaniasis and Sezary syndrome: endoscopic diagnosis]. AB - Sezary syndrome is a non-Hodgkin's lymphoma of cutaneous origin that provokes severe cellular immunosuppression leading to greater susceptibility to opportunistic infections. We present the case of a male patient with a diagnosis of Sezary syndrome complicated by visceral leishmaniasis and Mycobacterium avium complex coinfection, with intestinal involvement of both pathogens -an exceptional finding in the absence of HIV infection. The diagnosis was confirmed by bone marrow biopsy and oral endoscopy with intestinal biopsy. Because of the severity of the infection and the failure of conventional treatment, miltefosine, a new antiparasitic agent still under investigation, was administered with favorable response. However the patient developed fatal pneumonia. PMID- 17129550 TI - [Expert recommendations for the diagnosis and treatment of chronic hepatitis C infection in the prison setting]. AB - The prevalence of HCV infection in Spanish prisons is very high (38.5%). The characteristics of the infected patients, particularly the high rate of HIV coinfection, makes it very likely that the morbidity and mortality produced by serious liver disease secondary to this infection will increase considerably in the coming years. A group of Spanish experts with experience in patients who are inmates has been invited to establish a series of recommendations for the diagnosis and treatment of chronic hepatitis C infection in Spanish prisons. PMID- 17129551 TI - [Wilson's disease: forms of presentation in childhood]. AB - Wilson's disease can manifest with symptoms of liver disease or neuropsychiatric disorders in children and adults. This autosomal recessive disorder is caused by a copper metabolism disorder due to a mutation in the ATP7B transporter. Excessive amounts of copper accumulate in the body due to inhibition of the release of copper into bile. Because effective treatment is available, recognizing this disease in presymptomatic or early stages, when it can be reversed, is highly important. Diagnosis is often easy but the available tests (measurement of ceruloplasmin, and blood, urinary and liver copper levels) can be misleading. There is no single test with 100% sensitivity in screening nor do any of the tests, when used alone, provide 100% specificity. Diagnosis is currently based on the combination of clinical findings and the results of laboratory tests. Genetic study, with a finding of mutations in the two alleles of the ATP7B gene, is still not a rapid and easily available alternative and the absence of these mutations does not rule out the possible presence of other mutations not yet described. PMID- 17129552 TI - [Monitoring of thiopurine methyltransferase and thiopurine metabolites to optimize azathioprine therapy in inflammatory bowel disease]. AB - Determination of the activity of thiopurine methyltransferase (TPMT) and of thiopurine metabolites (6-thioguanine and 6-methylmercaptopurine nucleotides) could be useful for individualized monitoring of azathioprine (AZA) and 6 mercaptopurine (6-MP) doses. TPMT activity in the general population follows a trimodal distribution, in which approximately 0.3% of the population is homozygotic for the low-activity allele. A notable correlation has been observed between the low TPMP activity genotype or phenotype and the risk of myelotoxicity. Patients with a high TPMT activity genotype or homozygous phenotype should receive immunosuppressive doses that have clearly been demonstrated to be effective. In contrast, in patients with a low TPMT activity genotype or homozygous phenotype, the use of AZA/6-MP should be contraindicated or only very small doses should be administered. Importantly, TPMP deficiency explains only some cases of myelotoxicity and consequently periodic laboratory testing should be performed in patients receiving AZA/6-MP, even though TPMP function may be normal. Currently, the utility of routine thiopurine metabolite determinations in patients undergoing AZA/6-MP therapy has not been established and this practice should be limited to specific situations such as lack of response to thiopurine therapy or the occurrence of thiopurine-related adverse effects. Randomized trials comparing the routine strategy of AZA/6-MP dosing (based exclusively on the patient's weight) versus individualized monitoring (based on quantification of TPMP activity and/or thiopurine metabolites) are required before definitive conclusions on the most effective alternative can be drawn. PMID- 17129553 TI - [Endoscopic pancreatic sphincterotomy: when and how]. AB - Endoscopic pancreatic sphincterotomy (EPS) has fallen into disuse for some time because of the risk of severe complications. More recently, EPS has been advocated as an effective treatment modality for several pancreatic disorders, including severe chronic pancreatitis, pancreatic pseudocyst, ampulloma, pancreas divisum, and pancreatic sphincter dysfunction. Favorable outcomes in patients undergoing EPS to facilitate further interventions, in whom long-term follow-up was available, was 70%; complications occurred in 14% and reintervention was required in 23%. The results were as good as those of surgery after long-term follow-up. Patients who underwent some form of pancreatic drainage after sphincterotomy had fewer complications (p = 0.03). Approximately 75% of patients with pancreas divisum who presented with idiopathic acute recurrent pancreatitis improved after endoscopic therapy, but only 25% of patients experienced pain reduction of at least 50%. The National Institutes of Health Consensus recommends EPS in patients with type I sphincter of Oddi dysfunction (SOD). In patients with type II SOD, prior manometry should be performed. In our series of 17 patients, we obtained results similar to those of other studies, although the number of patients was small. EPS appears to be a safe and effective technique, but further, well-designed, multicenter, prospective and long-term studies are required to evaluate these results and settle current controversies. PMID- 17129554 TI - [Adult celiac disease and pseudoachalasia: a new clinical association]. PMID- 17129555 TI - [Prosthetic repair of perforation after endoscopic dilation in a patient with achalasia]. PMID- 17129556 TI - [Celiac disease and dilated cardiomyopathy]. PMID- 17129557 TI - [Ulcerative colitis and aplastic anemia]. PMID- 17129558 TI - [Spontaneous bacterial peritonitis due to Streptococcus constellatus]. PMID- 17129559 TI - [Two cases of segmental portal hypertension not associated with pancreatic disease]. PMID- 17129560 TI - A bivariate quantitative genetic model for a threshold trait and a survival trait. AB - Many of the functional traits considered in animal breeding can be analyzed as threshold traits or survival traits with examples including disease traits, conformation scores, calving difficulty and longevity. In this paper we derive and implement a bivariate quantitative genetic model for a threshold character and a survival trait that are genetically and environmentally correlated. For the survival trait, we considered the Weibull log-normal animal frailty model. A Bayesian approach using Gibbs sampling was adopted in which model parameters were augmented with unobserved liabilities associated with the threshold trait. The fully conditional posterior distributions associated with parameters of the threshold trait reduced to well known distributions. For the survival trait the two baseline Weibull parameters were updated jointly by a Metropolis-Hastings step. The remaining model parameters with non-normalized fully conditional distributions were updated univariately using adaptive rejection sampling. The Gibbs sampler was tested in a simulation study and illustrated in a joint analysis of calving difficulty and longevity of dairy cattle. The simulation study showed that the estimated marginal posterior distributions covered well and placed high density to the true values used in the simulation of data. The data analysis of calving difficulty and longevity showed that genetic variation exists for both traits. The additive genetic correlation was moderately favorable with marginal posterior mean equal to 0.37 and 95% central posterior credibility interval ranging between 0.11 and 0.61. Therefore, this study suggests that selection for improving one of the two traits will be beneficial for the other trait as well. PMID- 17129561 TI - Genetic analysis of growth curves using the SAEM algorithm. AB - The analysis of nonlinear function-valued characters is very important in genetic studies, especially for growth traits of agricultural and laboratory species. Inference in nonlinear mixed effects models is, however, quite complex and is usually based on likelihood approximations or Bayesian methods. The aim of this paper was to present an efficient stochastic EM procedure, namely the SAEM algorithm, which is much faster to converge than the classical Monte Carlo EM algorithm and Bayesian estimation procedures, does not require specification of prior distributions and is quite robust to the choice of starting values. The key idea is to recycle the simulated values from one iteration to the next in the EM algorithm, which considerably accelerates the convergence. A simulation study is presented which confirms the advantages of this estimation procedure in the case of a genetic analysis. The SAEM algorithm was applied to real data sets on growth measurements in beef cattle and in chickens. The proposed estimation procedure, as the classical Monte Carlo EM algorithm, provides significance tests on the parameters and likelihood based model comparison criteria to compare the nonlinear models with other longitudinal methods. PMID- 17129562 TI - Multibreed analysis by splitting the breeding values. AB - An equivalent model for multibreed variance covariance estimation is presented. It considers the additive case including or not the segregation variances. The model is based on splitting the additive genetic values in several independent parts depending on their genetic origin. For each part, it expresses the covariance between relatives as a partial numerator relationship matrix times the corresponding variance component. Estimation of fixed effects, random effects or variance components provided by the model are as simple as any model including several random factors. We present a small example describing the mixed model equations for genetic evaluations and two simulated examples to illustrate the Bayesian variance component estimation. PMID- 17129563 TI - Genetic heterogeneity of residual variance in broiler chickens. AB - Aims were to estimate the extent of genetic heterogeneity in environmental variance. Data comprised 99 535 records of 35-day body weights from broiler chickens reared in a controlled environment. Residual variance within dam families was estimated using ASREML, after fitting fixed effects such as genetic groups and hatches, for each of 377 genetically contemporary sires with a large number of progeny (>100 males or females each). Residual variance was computed separately for male and female offspring, and after correction for sampling, strong evidence for heterogeneity was found, the standard deviation between sires in within variance amounting to 15-18% of its mean. Reanalysis using log transformed data gave similar results, and elimination of 2-3% of outlier data reduced the heterogeneity but it was still over 10%. The correlation between estimates for males and females was low, however. The correlation between sire effects on progeny mean and residual variance for body weight was small and negative (-0.1). Using a data set bigger than any yet presented and on a trait measurable in both sexes, this study has shown evidence for heterogeneity in the residual variance, which could not be explained by segregation of major genes unless very few determined the trait. PMID- 17129564 TI - Comparison of methods for analysis of selective genotyping survival data. AB - Survival traits and selective genotyping datasets are typically not normally distributed, thus common models used to identify QTL may not be statistically appropriate for their analysis. The objective of the present study was to compare models for identification of QTL associated with survival traits, in particular when combined with selective genotyping. Data were simulated to model the survival distribution of a population of chickens challenged with Marek disease virus. Cox proportional hazards (CPH), linear regression (LR), and Weibull models were compared for their appropriateness to analyze the data, ability to identify associations of marker alleles with survival, and estimation of effects when all individuals were genotyped (full genotyping) and when selective genotyping was used. Little difference in power was found between the CPH and the LR model for low censoring cases for both full and selective genotyping. The simulated data were not transformed to follow a Weibull distribution and, as a result, the Weibull model generally resulted in less power than the other two models and overestimated effects. Effect estimates from LR and CPH were unbiased when all individuals were genotyped, but overestimated when selective genotyping was used. Thus, LR is preferred for analyzing survival data when the amount of censoring is low because of ease of implementation and interpretation. Including phenotypic data of non-genotyped individuals in selective genotyping analysis increased power, but resulted in LR having an inflated false positive rate, and therefore the CPH model is preferred for this scenario, although transformation of the data may also make the Weibull model appropriate for this case. The results from the research presented herein are directly applicable to interval mapping analyses. PMID- 17129565 TI - Genetic correlations between conformation traits and radiographic findings in the limbs of German Warmblood riding horses. AB - Studbook inspection (SBI) data of 20 768 German Warmblood mares and radiography results (RR) data of 5102 Hanoverian Warmblood horses were used for genetic correlation analyses. The scores on a scale from 0 to 10 were given for conformation and basic quality of gaits, resulting in 14 SBI traits which were used for the correlation analyses. The radiographic findings considered included osseous fragments in fetlock (OFF) and hock joints (OFH), deforming arthropathy in hock joints (DAH) and distinct radiographic findings in the navicular bones (DNB) which were analyzed as binary traits, and radiographic appearance of the navicular bones (RNB) which was analyzed as a quasi-linear trait. Genetic parameters were estimated multivariately in linear animal models with REML using information on 24 448 horses with SBI and/or RR records. The ranges of heritability estimates were h2 = 0.14-0.34 for the RR traits and h2 = 0.09-0.50 for the SBI traits. Negative additive genetic correlations of r(g) = -0.19 to 0.56 were estimated between OFF and conformation of front and hind limbs and walk at hand, and between DNB and hind limb conformation. There were indications of negative additive genetic correlations between DAH and all SBI traits, but because of low prevalence and low heritability of DAH, these results require further scrutiny. Positive additive genetic correlations of r(g) = 0.37-0.52 were estimated between OFF and withers height and between OFH and withers height, indicating that selection for taller horses will increase disposition to develop OFF and OFH. Selection of broodmares with regards to functional conformation will assist, but cannot replace possible selection against radiographic findings in the limbs of young Warmblood riding horses, particularly with regards to OFF. PMID- 17129566 TI - Sox genes in grass carp (Ctenopharyngodon idella) with their implications for genome duplication and evolution. AB - The Sox gene family is found in a broad range of animal taxa and encodes important gene regulatory proteins involved in a variety of developmental processes. We have obtained clones representing the HMG boxes of twelve Sox genes from grass carp (Ctenopharyngodon idella), one of the four major domestic carps in China. The cloned Sox genes belong to group B1, B2 and C. Our analyses show that whereas the human genome contains a single copy of Sox4, Sox11 and Sox14, each of these genes has two co-orthologs in grass carp, and the duplication of Sox4 and Sox11 occurred before the divergence of grass carp and zebrafish, which support the "fish-specific whole-genome duplication" theory. An estimation for the origin of grass carp based on the molecular clock using Sox1, Sox3 and Sox11 genes as markers indicates that grass carp (subfamily Leuciscinae) and zebrafish (subfamily Danioninae) diverged approximately 60 million years ago. The potential uses of Sox genes as markers in revealing the evolutionary history of grass carp are discussed. PMID- 17129567 TI - Short-term effects of countermeasures for improved safety and mobility at marked pedestrian crosswalks in Boras, Sweden. AB - The Swedish code concerning car drivers' responsibility to give way to pedestrians was strengthened in 2000. The primary aim of this study is to evaluate the short-term effects of the change in code. Another goal is to look at the effects of the reconstruction of four sites in Boras, Sweden. One site had changes made prior to the change of code, two test sites had countermeasures implemented during the study, and one comparison site was left unchanged. All the sites were chosen because schools were situated nearby. The focus of the evaluation was on children and elderly as pedestrians and cyclists. The goal of traffic calming of a 90 percentile driving speed below 30 km/h was not fulfilled at any of the test sites. A conclusion is that the height of a speed cushion is important. After the speed cushions were lowered from 70 mm to 55 mm, the 90 percentile speed increased from 34 km/h to 41 km/h. Sites with no speed cushions had much higher speeds. The design of an intersection influences road users' behavior. At the site where one crosswalk was removed, pedestrians that were using the remaining marked crosswalk were given way to less frequently than at the other sites. At intersections where most pedestrians used marked crosswalks, the children benefited the most in mobility. At the intersection where pedestrians used marked crosswalks to a lower extent after reconstruction, children and the elderly had the smallest increase in frequency of being given way to. After reconstruction to a court-yard street, the pedestrians were given way to a lower extent compared with the other sites, though the vehicle speeds were the lowest observed at this study. At the sites were no physical changes were made, the change of code improved driver yield behavior, but no more towards children than other age groups. PMID- 17129568 TI - Genetic structures of geographically distinct Plasmodium vivax populations assessed by PCR/RFLP analysis of the merozoite surface protein 3beta gene. AB - The recent resurgence of Plasmodium vivax malaria requires close epidemiological surveillance and monitoring of the circulating parasite populations. In this study, we developed a combination of polymerase chain reaction and restriction fragment length polymorphism (PCR/RFLP) method to investigate the genetic diversity of the P. vivax merozoite surface protein 3beta (PvMSP3beta) gene among four Asian parasite populations representing both tropical and temperate strains with dramatic divergent relapse patterns (N = 143). Using P. vivax field isolates from symptomatic patients, we have validated the feasibility of this protocol in distinguishing parasite genotypes. We have shown that PCR alone could detect three major size polymorphisms of the PvMSP3beta gene, and restriction analysis detected a total of 12 alleles within these Asian samples. Samples from different geographical areas differed dramatically in their PvMSP3beta allele composition and frequency, indicating that complex, yet different parasite genotypes were circulating in different endemic areas. This protocol allowed easy detections of multiple infections, which reached 20.5% in the samples from Thailand. It is interesting to note that samples from one temperate site in China collected during a recent outbreak of the disease also showed a high level of genetic diversity with multiple infections accounting for 5.6% of the samples. When combined with the PvMSP3alpha locus, this method provides better capability in distinguishing P. vivax genotypes and detecting mixed genotype infections. PMID- 17129569 TI - Detection of Osteopontin in the pericyst of human hepatic Echinococcus granulosus. AB - It aims at investigating the expression and distribution of the Osteopontin (OPN) in the pericyst of human hepatic Echinococcus granulosus and their related significances. Sixty pericysts excised by "sub-adventitial cystectomy" were studied. OPN was detected in 80% (48/60) of cysts by Western blotting and distributed in the side of "exocyst" layer directing to the parasite, also macrophages were identified in the vicinity of OPN by immunohistochemistry staining. The coexpression of OPN and CD68 was observed by immunofluorescence double labeling and analyzed by Image-Pro Plus 5.1; with special stain techniques, variable degrees of calcium deposits were observed in 80% (48/60) cysts, and the calcium deposits concurrencely found with the OPN expression. The selective distribution of OPN, calcium in the "exocyst" provides a new pathological evidence for the "sub-adventitial cystectomy" we developed. The pericyst of hepatic E. granulosus consists of two detachable layers with different formative mechanisms: the "exocyst" layer directing towards the cyst of parasite was the result of granulomatous reaction; also the results suggest OPN is one regulator in the granulomatous reaction and calcification of "exocyst". PMID- 17129570 TI - Enhanced fluorescence resonance energy transfer immunoassay with improved sensitivity based on the Fab'-based immunoconjugates. AB - Fluorescence resonance energy transfer (FRET) is a powerful technique to monitor protein-protein interaction. Recently, we developed homogeneous and noncompetitive immunoassay based on the enhanced FRET by leucine zipper interaction. Here we improved the assay by establishing a general method for preparation of the Fab'-based immunoconjugate. Anti-human serum albumin Fab' numbers 11 and 13 were chemically conjugated with recombinant proteins consisting of thioredoxin, flexible linker, and green fluorescent protein color variant tethered with a leucine zipper motif. Compared with single chain antibody variable region-based fusion proteins prepared by the gene fusion method in our previous study, the resultant Fab'-based immunoconjugates accomplished an assay with nearly 10 times greater sensitivity. Furthermore, the conjugation method enabled us to apply the assay generally to measurement of another high-molecular weight antigen for which antibodies prepared for sandwich immunoassay are commercially available. Because of the facility and generality of the preparation method for the immunoconjugate, the assay is expected to be applied to many antigens that require rapid diagnosis and moderate measurement range. PMID- 17129571 TI - Large systematic errors compromise quantitation of intrinsically unstructured proteins. PMID- 17129572 TI - Mixing two differently labeled substrates in one immobilized metal assay for phosphochemicals assay to improve data quality. PMID- 17129573 TI - Functional roles of oral reflexes in chewing and biting: phase-, task- and site dependent reflex sensitivity. AB - Because loading during chewing is not totally predictable and jaw-closing muscles are strong and act over short distances, feedback from oral receptors is important in the control of mastication. Information on such feedback can be obtained by studying reflexes in jaw muscle EMGs. This review will deal with the contribution of reflex mechanisms to modifying motor neuron activity during chewing, and the dependency of reflex sensitivity on motor task, phase of movement, and site of stimulation. PMID- 17129574 TI - Regulation of Ca2+ mobilization by prolactin in mammary gland cells: possible role of secretory pathway Ca2+-ATPase type 2. AB - Regulatory role of prolactin (PRL) on Ca2+ mobilization in human mammary gland cell line MCF-7 was examined. Direct addition of PRL did not affect cytoplasmic Ca2+ concentration ([Ca2+]i); however, treatment with PRL for 24h significantly decreased the peak level and duration time of [Ca2+]i elevation evoked by ATP or thapsigargin (TG). Intracellular Ca2+ release by IP3 or TG in permeablized cells was not decreased after PRL-treatment, indicating that the Ca2+ release was not impaired by PRL treatment. Extracellular Ca2+ entry evoked by ATP or TG was likely to be intact, because entry of extracellular Ba2+ was not affected by PRL treatment. Among Ca2+-ATPases expressed in MCF-7 cells, we found significant increase of secretory pathway Ca2+-ATPase type 2 (SPCA2) mRNA in PRL-treated cells by RT-PCR experiments including quantitative RT-PCR. Knockdown of SPCA2 by siRNA in PRL-treated cells showed similar Ca2+ mobilization to that in PRL untreated cells. The present results suggest that PRL facilitates Ca2+ transport into Golgi apparatus and may contribute the supply of Ca2+ to milk. PMID- 17129575 TI - High molecular weight adiponectin reduces apolipoprotein B and E release in human hepatocytes. AB - Low circulating levels of high molecular weight adiponectin (HMW-Apm) have been linked to dyslipidaemia and systemic HMW-Apm negatively correlates with very low density lipoprotein (VLDL), apolipoprotein B (ApoB), and ApoE and is positively associated with ApoA-I. Therefore, it was investigated whether HMW-Apm alters the hepatic synthesis of ApoB, ApoE, and ApoA-I or the activity of the hepatic ATP binding cassette transporter A1 (ABCA1), as the main determinant of plasma HDL. HMW-Apm reduces hepatic ApoB and ApoE release whereas ABCA1 protein, activity and ApoA-I were not altered. Global gene expression analysis revealed that hepatic nuclear factor 4-alpha (HNF4-alpha) and HNF4-alpha regulated genes like ApoB are downregulated by HMW-Apm and this was confirmed at the mRNA and protein level. Therefore it is concluded that HMW-adiponectin may ameliorate dyslipidaemia by reducing the hepatic release of ApoB and ApoE, whereas ABCA1 function and ApoA-I secretion are not influenced. PMID- 17129576 TI - Identification of differentially expressed genes in omental adipose tissues of obese patients by suppression subtractive hybridization. AB - To identify differentially expressed genes between obese individuals and normal control, we have undertaken suppression subtractive hybridization (SSH). Omental adipose tissues were obtained via abdominal surgery for appendicitis in both 13 obese subjects [BMI (body mass index) >30 kg/m2] and 13 normal subjects (BMI >18 and <25 kg/m2). Following SSH, about one thousand clones were sequenced and found to derive from 426 different genes. These predominately expressed genes included genes involved in lipid metabolism, cytokines, signal transduction, GLUT4 translocation, cell cycle and growth, cytoskeleton, and others. Although more detailed analyses are necessary, it is anticipated that further study of genes identified will provide insights into their specific roles in the etiology of obesity. PMID- 17129577 TI - Histamine H3 receptor antagonists: from target identification to drug leads. AB - The successful cloning and functional expression of the histamine H(3) receptor in the late 1990 s has greatly facilitated our efforts to identify small molecule, non-imidazole based compounds to permit the evaluation of H(3) antagonists in models of CNS disorders. High-throughput screening identified several series of lead compounds, including a series of imidazopyridines, which led to JNJ-6379490, a compound with high affinity for the human H(3) receptor. Analysis of structural features common to several series of non-imidazole H(3) receptor ligands resulted in a pharmacophore model. This model led to the design of JNJ-5207852, a diamine-based H(3) antagonist with good in vitro and in vivo efficacy but with an undesirable long half-life. However, further modifications of the template provided an understanding of the effect of structural modifications on pharmacokinetic properties, ultimately affording several additional series of compounds including JNJ-10181457, a compound with an improved pharmacokinetic profile. These compounds allowed in vivo pharmacological evaluation to show that H(3) antagonists promote wakefulness, but unlike modafinil and classical psychostimultants, they do not increase locomotor activity or produce any alteration of the EEG power spectral activity in rats. H(3) antagonists also increase extracellular acetylcholine and norepinephrine but not dopamine in rat frontal cortex and show efficacy in various models of learning-memory deficit. In addition, cFos immunoreactivity studies show H(3) antagonists activate neuronal cells in restricted rat brain regions in contrast to widespread activation after modafinil or amphetamine treatment. Therefore, H(3) antagonists are promising clinical candidates for the treatment of excessive day time sleepiness and/or cognitive disorders. PMID- 17129578 TI - Phospholipase C-dependent control of cardiac calcium homeostasis involves a TRPC3 NCX1 signaling complex. AB - OBJECTIVE: Members of the classical transient receptor potential protein (TRPC) family are considered as key components of phospholipase C (PLC)-dependent Ca2+ signaling. Previous results obtained in the HEK 293 expression system suggested a physical and functional coupling of TRPC3 to the cardiac-type Na+/Ca2+ exchanger, NCX1 (sodium calcium exchanger 1). This study was designed to test for expression of TRPC3 (transient receptor potential channel 3) and for the existence of a native TRPC3/NCX1 signaling complex in rat cardiac myocytes. METHODS: Protein expression and cellular distribution were determined by Western blot and immunocytochemistry. Protein-protein interactions were investigated by reciprocal co-immunoprecipitation and glutathione S-transferase (GST)-pulldown experiments. Recruitment of protein complexes into the plasma membrane was assayed by surface biotinylation. The functional role of TRPC3 was investigated by fluorimetric recording of angiotensin II-induced calcium signals employing a dominant negative knockdown strategy. RESULTS: TRPC3 immunoreactivity was observed in surface plasma membrane regions and in an intracellular membrane system. Co immunolabeling of TRPC3 and NCX1 indicated significant co-localization of the two proteins. Both co-immunoprecipitation and GST-pulldown experiments demonstrated association of TRPC3 with NCX1. PLC stimulation was found to trigger NCX-mediated Ca2+ entry, which was dependent on TRPC3-mediated Na+ loading of myocytes. This NCX-mediated Ca2+ signaling was significantly suppressed by expression of a dominant negative fragment of TRPC3. PLC stimulation was associated with increased membrane presentation of both TRPC3 and NCX1. CONCLUSION: These results suggest a PLC-dependent recruitment of a TRPC3-NCX1 complex into the plasma membrane as a pivotal mechanism for the control of cardiac Ca2+ homeostasis. PMID- 17129579 TI - Study of antimutagenic and antioxidant activities of gallic acid and 1,2,3,4,6 pentagalloylglucose from Pistacia lentiscus. Confirmation by microarray expression profiling. AB - In vitro antioxidant and antimutagenic activities of two polyphenols isolated from the fruits of Pistacia lentiscus was assessed. Antioxidant activity was determined by the ability of each compound to scavenge the free radical 1,1 diphenyl-2-picrylhydrazyl (DPPH*), to inhibit xanthine oxidase and to inhibit the lipid peroxidation induced by H(2)O(2) in K562 cell line. Antimutagenic activity was assayed with SOS chromotest using Escherichia coli PQ37 as tester strain and Comet assay using K562 cell line. 1,2,3,4,6-Pentagalloylglucose was found to be more effective to scavenge DPPH* radical and protect against lipid peroxidation. Moreover, these two compounds induced an inhibitory activity against nifuroxazide and aflatoxin B1 mutagenicity. The protective effect exhibited by these molecules was also determined by analysis of gene expression as response to an oxidative stress. For this purpose, we used a cDNA-microarray containing 82 genes related to cell defense, essentially represented by antioxidant and DNA repair proteins. We found that 1,2,3,4,6-pentagalloylglucose induced a decrease in the expression of 11 transcripts related to antioxidant enzymes family (GPX1, TXN, AOE372, SHC1 and SEPW1) and DNA repair (POLD1, APEX, POLD2, MPG, PARP and XRCC5). The use of Gallic acid, induced expression of TXN, TXNRD1, AOE372, GSS (antioxidant enzymes) and LIG4, POLD2, MPG, GADD45A, PCNA, RPA2, DDIT3, HMOX2, XPA, TDG, ERCC1 and GTF2H1 (DNA repair) as well as the repression of GPX1, SEPW1, POLD1 and SHC1 gene expression. PMID- 17129581 TI - The role of functional information for infant categorization. AB - This report examines whether knowledge about function influences the formation of artifact categories in 11-12- month old infants. Using an object-examination task, a set of artificial stimuli was presented that could either be grouped according to overall similarity or according to similarity in one functionally relevant part. Experiment 1 revealed that infants categorized the objects according to overall similarity but not part similarity under control conditions. Experiment 2 showed that after having seen the experimenter demonstrating the functional use of the critical part, infants later categorized the stimuli according to part similarity. When the same actions were performed without producing any effect, infants failed to categorize according to the critical part. This set of findings suggests that 11-12-month old infants use functional information as a cue to categorization. PMID- 17129580 TI - The association of PC-1 (ENPP1) K121Q polymorphism with metabolic syndrome in patients with coronary heart disease. AB - BACKGROUND: Metabolic syndrome (MS) is a clinical feature, closely associated with insulin resistance, one of the prime underlying causes of overall cardiovascular morbidity, including coronary heart disease (CHD). Considering the association between PC-1 121Q genotype and insulin resistance phenotype, the aim of the present study was to investigate the contribution of PC-1 K121Q polymorphism to the development of MS and its concomitant disorders in CHD patients. METHODS: A total of 130 Caucasians from Serbia, including 80 CHD patients (aged 59.4+/-8.6 years, of a mean BMI 28.9+/-3.9 kg/m2) and 50 control subjects (aged 48.0+/-6.4 years, of a mean BMI 29.6+/-2.1 kg/m2), were genotyped for PC-1 K121Q using a mutagenic separated PCR assay, in order to determine the prevalence of the PC 121Q variant in individuals suffering from CHD and its association with MS. RESULTS: The frequency of PC-1 121Q allele found in CHD patients was 28.5%, with significantly (P<0.01) higher prevalence in those with MS (40% vs. 10%). Both MS (P<0.01) and its components [central obesity (P<0.01), low HDL-cholesterol (P<0.01) and high triglycerides (P<0.05)] were significantly more prevalent in CHD 121Q carriers compared to CHD patients who exhibited the wild-type genotype. A binary logistic regression model has revealed that PC-1 121Q allele carriers had a 5.5 fold increased odds (95%CI: 1.4-20.9, P=0.01) for the MS compared to wild-type carriers. The PC-1 121Q allele contributed to MS components as well, although these associations did not reach statistical significance. CONCLUSION: The findings of the present study support the hypothesis that the PC-1 (ENPP1) 121Q allele is associated with the genetic susceptibility for MS in patients with CHD. Further studies and more extensive research in this area are needed, not only to confirm this association, but to elucidate it in more details. PMID- 17129582 TI - Similarity classifier using similarity measure derived from Yu's norms in classification of medical data sets. AB - A new approach using a similarity measure based on Yu's norms is presented for the detection of erythemato-squamous diseases, diabetes, breast cancer, lung cancer and lymphography. The domain contains records of patients with known diagnoses. The results are very promising with all data sets and (in conclusion, can be drawn that) a similarity model derived from Yu's norms could be used for the diagnosis of patients taking into consideration the error rate. A similarity classifier derived from Yu's norms was used to detect the six erythemato-squamous diseases when 34 features defining six disease indications were used as inputs. The results confirmed that the proposed model has potential in detecting the erythemato-squamous diseases. The similarity model derived from Yu's norms achieved an accuracy rate (97.8%) which was higher than that of the stand-alone neural network model or the ANFIS model suggested in Ubeyli and Guler [Automatic detection of erythemato-squamous diseases using adaptive neuro-fuzzy inference systems, Comput. Biol. Med. 35 (2005) 421-433] or the similarity model based on Lukasiewicz similarity [Luukka and Leppalampi, Similarity classifier with generalized mean applied to medical data, Comput. Biol. Med. 36 (2006) 1026 1040]. With PIMA Indian diabetes, the detection model has an error rate of about 24% which is much better than the overall rate of 33% for diabetes. Also, a classifier was applied to the lung cancer data set and the results were to my knowledge better than before. When the lung cancer data were preprocessed with an entropy minimization technique and the classifier with similarity based on Yu's norm was applied, 99.99% accuracy was achieved. The use of this preprocessing method enhanced the results over 30%. In lymphography, entropy minimization also enhanced the results remarkably and 86.2% accuracy was achieved. PMID- 17129583 TI - "Awake" laparoscopy for the evaluation of equivocal penetrating abdominal wounds. AB - BACKGROUND: Diagnostic laparoscopy is useful for the assessment of equivocal penetrating abdominal wounds, and has become the modality of choice for the evaluation of such wounds at our institution. We hypothesised that, in appropriate patients, diagnostic "awake" laparoscopy (AL) could be performed under local anaesthesia in the emergency department (ED), allowing for expedited discharge and potential cost savings. METHODS: Selected haemodynamically stable patients with penetrating abdominal injury underwent AL. Suitability for AL was at the discretion of the attending surgeon. Identification of peritoneal penetration by AL led to exploratory laparotomy in the operating room. Patients with no evidence of peritoneal penetration were discharged from the ED (ALneg). These patients were matched to a cohort of 24 patients who underwent diagnostic laparoscopy in the OR which was negative for peritoneal penetration (DLneg). Length of stay and hospital charges were compared. RESULTS: Over a 30-month period, 15 patients underwent AL without complication. No peritoneal penetration was found in 11 patients. The remaining four patients underwent exploratory laparotomy, of which two were positive for intra-abdominal injury. Mean time to discharge was 7h in the ALneg group versus 18 h in the DLneg group (p=0.0003). Cost savings on hospital charges averaged 2227 US dollars per patient in the ALneg group compared with the DLneg group. CONCLUSIONS: AL may be safely performed in the ED, allowing for expedited patient discharge. Cost savings are achieved by the avoidance of charges inherent to diagnostic laparoscopy performed in the operating room. PMID- 17129584 TI - Recent developments in liquid chromatography--impact on qualitative and quantitative performance. AB - In order to reduce the analysis time and maintain good efficiency in liquid chromatography (LC), several solutions are currently being investigated. The focus of this study was to compare, both qualitatively and quantitatively, the chromatographic performance of a conventional LC with selected approaches, namely monolithic supports, high temperature LC (up to 90 degrees C), and sub-2 microm particles combined with high pressure (up to 1000 bar). This comparison was achieved from a qualitative point of view with a special attention paid to the analysis of time reduction, efficiency improvement, and pressure constraint. For this purpose, the different approaches were discussed using Knox curves and other kinetic plots. It appeared that columns packed with sub-2 microm particles under high-pressure conditions (UPLC) were well adapted and this option represents an attractive alternative to conventional LC; however, the other alternative approaches should not be neglected. The quantitative evaluation of these techniques was performed on the basis of the validation of results of a pharmaceutical formulation (Rapidocaine), following SFSTP 2003 guidelines. Fast LC approaches demonstrated equivalent performance to conventional LC in terms of trueness, precision, and accuracy profile, with a significant time reduction (up to 8x) according to the selected methodology. PMID- 17129585 TI - Pressure-driven flow control system for nanofluidic chemical process. AB - We developed a novel flow control system for a nanofluidic chemical process. Generally, flow control in nanochannels is difficult because of its high-pressure loss with very small volume flow rate. In our flow control method, liquid pressure in a microchannel connected to the nanochannels is regulated by utilizing a backpressure regulator. The flow control method was verified by using simple structured microchip, which included parallel nanochannels. We found that the observed flow rate was three times lower than the value expected from Hagen Poiseuille's equation. That implied a size-dependent viscosity change in the nanochannels. Then, we demonstrated mixing of two different fluorescent solutions in a Y-shaped nanochannel and also a proton exchange reaction in the Y-shaped nanochannel. The flow control method will contribute to further integration of nanochemical systems. PMID- 17129586 TI - Purification of monocytes from cryopreserved mobilized apheresis products by elutriation with the Elutra device. AB - The Elutra biomedical device allows semi-automatic enrichment of monocytes by elutriation, using a single-use, closed and cGMP compliant tubing set, in a cost effective way. The procedure has been validated using fresh apheresis products from nonmobilized donors. We here evaluated the possibility of using Elutra to enrich monocytes from frozen/thawed apheresis products collected from mobilized healthy donors. Frozen apheresis products from 6 G CSF mobilized donors were thawed and used in 16 elutriation procedures. We compared the recovery and purity of enriched monocytes using different buffer compositions and elutriation profiles. Elutriated monocytes were cultured to generate mature dendritic cells (DCs). Depending in part of the initial granulocyte contamination in the apheresis product, the use of Desoxyribo Nuclease (DNAse) to avoid aggregation, was needed through only the initial steps or throughout the elutriation process. The average monocyte recovery was 85+/-31%. The average purity was 73+/-9%. The recovery of mature DC at d8 of culture was 20+/-6% of the input monocyte numbers. We conclude that Elutra allows the purification of monocytes from thawed mobilized apheresis. It requires no pre-processing of the cell product before elutriation, and allows the generation of phenotypically mature DC in quantities that are compatible with a clinical use. PMID- 17129587 TI - Catecholamines, infection, and death in acute ischemic stroke. AB - Experimental studies have recently suggested that acute ischemia may facilitate the appearance of fatal infections as part of a brain-induced immunodepression syndrome. However, the mechanisms and neurological consequences of infections complicating acute ischemic stroke have received much less attention at the bedside. The incidence of infection and death after non-septic stroke was compared in this prospective study with longitudinal changes of cytokines, leukocytes, normetanephrine (NMN) and metanephrine (MN) in 75 consecutive patients. In multivariate analysis, infection, n = 13 (17%), was associated with the upper quartile of MN (OR 3.51, 95% CI 1.30-9.51), neurological impairment (NIHSS) on admission (OR 3.99, 95% CI 1.34-11.8), monocyte count (OR 1.78, 95% CI 1.13-2.79), and increased interleukin (IL)-10 (OR 1.54, 95% CI 1.00-2.38). Mortality at 3 months, n = 16 (21%), was associated with increased levels of NMN on admission (OR 2.34 95% CI 1.15-4.76), NIHSS score (OR 2.57, 95% CI 1.29-5.11), and higher IL-6 levels (OR 1.29, 95% 1.00-1.67). These findings suggest that acute ischemic stroke is associated with an early activation of the sympathetic adrenomedullar pathway that lowers the threshold of infection and increases the risk of death. Moreover, these findings are independent of the blood borne effects of pro- and anti-inflammatory cytokines, and circulating leukocytes. PMID- 17129588 TI - Dibenzocyclooctadiene lignans: a class of novel inhibitors of multidrug resistance-associated protein 1. AB - We recently reported that dibenzocyclooctadiene lignans were a novel class of P glycoprotein (P-gp) inhibitors. In this study, we demonstrated that the lignans of this class were also effective inhibitors of multidrug resistance-associated protein 1 (MRP1). The activities of 5 dibenzocyclooctadiene lignans (schisandrin A, schisandrin B, schisantherin A, schisandrol A, and schisandrol B) to reverse MRP1-mediated drug resistance were tested using HL60/Adriamycin (ADR) and HL60/Multidrug resistance-associated protein (MRP), two human promyelocytic leukemia cell lines with overexpression of MRP1 but not P-gp. The five lignans could effectively reverse drug resistance of the two cell lines to vincristine, daunorubicin, and VP-16. This study, together with our previous reports, proves that dibenzocyclooctadiene lignans have multiple activities against cancer multidrug resistance, including inhibition of P-gp and MRP1, and enhancement of apoptosis. Considering that cancer multidrug resistance (MDR) is multifactorial, agents with broad activities are preferable to the use of combination of several specific modulators to prevent drug-drug interaction and cumulative toxicity. PMID- 17129589 TI - Ginsenoside Rb1 promotes adipogenesis in 3T3-L1 cells by enhancing PPARgamma2 and C/EBPalpha gene expression. AB - Evidence has accumulated that ginseng and its main active constituents, ginsenosides, possess anti-diabetic and insulin-sensitizing properties which may be partly realized by regulating adipocyte development and functions. In the present study, we explored the effect of ginsenoside Rb(1), the most abundant ginsenoside in ginseng root, on adipogenesis of 3T3-L1 cells. We found that with standard differentiation inducers, ginsenoside Rb(1) facilitated adipogenesis of 3T3-L1 preadipocytes in a dose-dependent manner; 10 microM Rb(1) increased lipid accumulation by about 56%. Treatment of differentiating adipocytes with 10 microM Rb(1) increased the expression of mRNA and protein of PPARgamma(2) and C/EBPalpha, as well as mRNA of ap2, one of their target genes. After the treatment of differentiating adipocytes with Rb(1), basal and insulin-mediated glucose uptake was significantly augmented, accompanied by the up-regulation of mRNA and protein level of GLUT4, but not of GLUT1. In addition, ginsenoside Rb(1) also inhibited the proliferation of preconfluent 3T3-L1 preadipocytes. Our data indicate that anti-diabetic and insulin-sensitizing activities of ginsenosides, at least in part, are involved in the enhancing effect on PPARgamma2 and C/EBPalpha expression, hence promoting adipogenesis. PMID- 17129590 TI - 2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potentiates nitrosation of a heterocyclic amine carcinogen by nitric oxide. AB - Although nitrosation plays an important role in initiation of carcinogenesis, the reactive nitrogen oxygen species (RNOS) mediating this reaction by multiple pathways have not been determined. The heterocyclic amine carcinogen 2-amino-3 methylimidazo[4,5-f]quinoline (IQ) was used as a target to investigate RNOS and pathways for potentiation of nitric oxide (NO)-mediated nitrosation. 2-(4 Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (CPTIO) oxidizes NO to NO(2)(.) and was used as a tool to investigate NO(2)(.) potentiation of nitrosation. The IQ nitrosation product, 2-nitrosoamino-3-methylimidazo[4,5 f]quinoline ((14)C-N-NO-IQ), was monitored by HPLC. Autoxidation of NO, generated by spermine NONOate (2.4 microM NO/min) for 7.5 min, did not convert 10 microM (14)C-IQ to N-NO-IQ. However, the presence of 15 muM CPTIO resulted in 3 microM N NO-IQ formation. Potentiation by CPTIO occurred at low and high fluxes of NO, 0.075 to 1.2 microM/min, and over a range of IQ to CPTIO ratios of 0.5 to 10. A significant portion of N-NO-IQ formation was insensitive to azide (10 mM) inhibition, suggesting oxidative nitrosylation. NADH (0.02 mM) did not alter nitrosation by autoxidation, but effectively inhibited potentiation by CPTIO. Ascorbic acid (0.2 mM) and 5,5-dimethyl-1-pyrroline N-oxide (30 mM) inhibited nitrosation with or without CPTIO, while superoxide dismutase was not inhibitory. The RNOS produced by CPTIO had a 27-fold greater affinity for IQ than those produced by autoxidation. Results are consistent with NO(2)(.) or a RNOS like NO(2)(.) potentiating IQ oxidative nitrosylation. Nitrosation occurring at both low and high fluxes of NO can contribute to carcinogenesis. PMID- 17129591 TI - Fractionation of memory in medial temporal lobe amnesia. AB - We report a comprehensive investigation of the anterograde memory functions of two patients with memory impairments (RH and JC). RH had neuroradiological evidence of apparently selective right-sided hippocampal damage and an intact cognitive profile apart from selective memory impairments. JC, had neuroradiological evidence of bilateral hippocampal damage following anoxia due to cardiac arrest. He had anomic and "executive" difficulties in addition to a global amnesia, suggesting atrophy extending beyond hippocampal regions. Their performance is compared with that of a previously reported hippocampal amnesic patient who showed preserved recollection and familiarity for faces in the context of severe verbal and topographical memory impairment [VC; Cipolotti, L., Bird, C., Good, T., Macmanus, D., Rudge, P., & Shallice, T. (2006). Recollection and familiarity in dense hippocampal amnesia: A case study. Neuropsychologia, 44, 489-506.] The patients were administered experimental tests using verbal (words) and two types of non-verbal materials (faces and buildings). Receiver operating characteristic analyses were used to estimate the contribution of recollection and familiarity to recognition performance on the experimental tests. RH had preserved verbal recognition memory. Interestingly, her face recognition memory was also spared, whilst topographical recognition memory was impaired. JC was impaired for all types of verbal and non-verbal materials. In both patients, deficits in recollection were invariably associated with deficits in familiarity. JC's data demonstrate the need for a comprehensive cognitive investigation in patients with apparently selective hippocampal damage following anoxia. The data from RH suggest that the right hippocampus is necessary for recollection and familiarity for topographical materials, whilst the left hippocampus is sufficient to underpin these processes for at least some types of verbal materials. Face recognition memory may be adequately subserved by areas outside of the hippocampus. PMID- 17129592 TI - Comparative study of the quality and efficacy of originator and generic albendazole for mass treatment of soil-transmitted nematode infections in Nepal. AB - The quality and efficacy of two locally manufactured generic albendazole (ABZ) products (Curex and Royal Drug) used for deworming children in Nepal since 1999 were tested against the originator product (GlaxoSmithKline (GSK)). The study included disintegration and dissolution testing according to the Indian Pharmacopoeia (IP) and the United States Pharmacopeia (USP), respectively, as well as a randomised controlled clinical trial comparing cure rates (CR) and egg reduction rates (ERR) for Ascaris lumbricoides, Trichuris trichiura and hookworm infections. Stool samples from 1277 children were examined before and 21 days after treatment. For A. lumbricoides, GSK (97.0%) and Royal Drug (95.0%) ABZ achieved significantly higher CRs than Curex ABZ (82.6%); however, all products achieved ERRs >90%. For T. trichiura, Curex ABZ showed significantly lower ERRs (63.2%). For hookworms, GSK ABZ performed significantly better (CR 74.3%, ERR 87.1%) than Royal Drug ABZ (CR 53.3%, ERR 80.8%) and Curex ABZ (CR 50.7%, ERR 73.1%). Only the GSK product passed both disintegration and dissolution tests according to the IP and USP. Both generic products failed the dissolution tests. Curex ABZ showed poor disintegration. Despite its lower efficacy, the cheaper Curex product achieved good results in controlling morbidity due to soil transmitted helminth infections. This study shows that the cost effectiveness of drugs used in mass deworming campaigns should not be inferred on the basis of a single quality testing parameter. PMID- 17129593 TI - Motion-onset VEPs: characteristics, methods, and diagnostic use. AB - This review article summarises the research on the motion-onset visual evoked potentials (VEPs) and important motion stimulus parameters which have been clarified. For activation of the visual motion processing system and evocation of the motion-onset specific N2 peak (with latency of 160-200ms) from the extra striate temporo-occipital and/or parietal cortex, the following stimulus parameters can be recently recommended: low luminance (Nic or Sal-->Sal) or a different state (Nic-->Sal or Sal- >Nic) after acquiring the conditioned response. A state-dependency account predicts disruption in conditioned goal tracking for rats that receive a shift in drug state on the test day. This disruption did not occur suggesting that differential control of conditioned responding by nicotine is more likely due to a direct excitatory association between the interoceptive cueing effects of nicotine and the appetitive qualities of sucrose. PMID- 17129620 TI - Molecular cloning and characterization of a C-type lectin from Ancylostoma ceylanicum: evidence for a role in hookworm reproductive physiology. AB - Lectins comprise a family of related proteins that mediate essential cell functions through binding to carbohydrates. Within this protein family, C-type lectins are defined by the requirement of calcium for optimal biologic activity. Using reverse transcription PCR, a cDNA corresponding to a putative C-type lectin has been amplified from the hookworm parasite Ancylostoma ceylanicum. The 550 nucleotide open reading frame of the A. ceylanicum C-type Lectin-1 (AceCTL-1) cDNA corresponds to a 167 amino acid mature protein (18,706 Da) preceded by a 17 amino acid secretory signal sequence. The recombinant protein (rAceCTL-1) was expressed in Drosophila S2 cells and purified using a combination of affinity chromatography and reverse phase HPLC. Using in vitro carbohydrate binding studies, it was determined that rAceCTL-1 binds N-acetyl-d-glucosamine, a common component of eukaryotic egg cell membranes. Using a polyclonal IgG raised against the recombinant protein, the native AceCTL-1 was identified in sperm and soluble protein extracts of adult male A. ceylanicum by immunoblot. Probing of adult hookworm sections with the polyclonal IgG demonstrated localization to the testes in males, as well as the spermatheca and developing embryos in females, consistent with its role as a sperm protein. Together, these data strongly suggest that AceCTL-1 is a male gender-specific C-type lectin with a function in hookworm reproductive physiology. PMID- 17129621 TI - Does day care also provide care for the caregiver? AB - People caring for relatives with dementia may need respite to be able to cope and maintain some quality of life. With the aim to investigate whether day care is an effective form of respite for caregivers of dementia sufferers, 115 new day care clients were invited and 90 caregivers agreed to participate. Of these, 51 fulfilled the inclusion criteria and were asked to assess feelings of worry, depression, overload and role captivity before using day care and again 4 months later. The study group comprised both co-residing (CR) and non-co-residing (NCR) caregivers and care recipients. Those NCR caregivers whose elderly relative dropped out of day care had significantly higher values for worry, overload and role captivity, and a higher level of depression than the caregivers in the NCR group whose care recipient continued day care. Although this study is of an explorative character, some conclusions may be drawn from it. Dementia sufferers seem to need an informal caregiver to help them when they are not at day care. Day care is probably a form of support most suitable for caregivers, who are looking after relatives in the early stages of the illness and are not yet too exhausted. PMID- 17129622 TI - Animal-based national surveillance for zoonotic disease: quality, limitations, and implications of a model system for monitoring rabies. AB - Surveillance for zoonotic diseases among wildlife is a research and public health challenge. The inherent limitations posed by the requisite human-animal interactions are often undefined and underappreciated. The national surveillance system for animal rabies in the United States was examined as a model system; reporting of animal rabies is legally mandated, each case of rabies is laboratory confirmed, and data have been consistently collected for more than 50 years. Factors influencing the monthly counts of animal rabies tests reported during 1992-2001 were assessed by univariate and multivariable regression methods. The suitability of passively collected surveillance data for determining the presence or absence of the raccoon-associated variant of rabies within states and within individual counties was assessed by determining critical threshold values from the regression analyses. The size of the human population and total expenditures within a county accounted for 72% and 67%, respectively, of the variance in testing. The annual median number of rabies tests performed was seven for counties without rabies, 22 for counties with non-raccoon rabies, and 34 for counties with raccoon rabies. Active surveillance may be required in locales with sparse human populations when a high degree of confidence in the status of rabies is required. PMID- 17129623 TI - The earnings of U.S. health economists. AB - This paper presents data from a 2005 survey of health economists who were U.S. members of the International Health Economics Association or the Health Economics Special Interest Group of Academy Health. We present summary statistics of health economist earnings by rank and type of employer, estimate log earnings models as a function of education, experience, type of employer, and research productivity, and provide summary statistics of starting salaries expected to be offered to new Ph.D. health economists by type of employer. Our findings are compared to those for economics departments, business schools, schools of public health, and for health services researchers. PMID- 17129624 TI - Effects of infertility insurance mandates on fertility. AB - Infertility currently affects over 6 million individuals in the United States. While most health insurance plans nationwide do not cover infertility diagnoses or treatments, to date 15 states have enacted some form of infertility insurance mandate. In this paper, I use data from the Vital Statistics Detail Natality Data and Census population estimates to examine whether these state-level mandates were successful in increasing fertility rates. Using a difference-in-differences approach, I exploit variation in the enactment of mandates both across states and over time, and identify treatment and control groups that should have been differentially affected by infertility coverage. My results suggest that the mandates significantly increase first birth rates for women over 35, and these results are robust to a number of specification tests. PMID- 17129625 TI - Insurance choice and tax-preferred health savings accounts. AB - We develop an infinite horizon utility maximization model of the interaction between insurance choice and tax-preferred health savings accounts. The model can be used to examine a wide range of policy options, including flexible spending accounts, health savings accounts, and health reimbursement accounts. We also develop a 2-period model to simulate various implications of the model. Key results from the simulation analysis include the following: (1) with no adverse selection, use of unrestricted health savings accounts leads to modest welfare gains, after accounting for the tax revenue loss; (2) with adverse selection and an initial pooling equilibrium comprised of "sick" and "healthy" consumers, introducing HSAs can, but does not necessarily, lead to a new pooling equilibrium. The new equilibrium results in a higher coinsurance rate, an increase in expected utility for healthy consumers, and a decrease in expected utility for sick consumers; (3) with adverse selection and a separating equilibrium, both sick and healthy consumers are better off with a health savings account; (4) efficiency gains are possible when insurance contracts are explicitly linked to tax-preferred health savings accounts. PMID- 17129627 TI - Automated alkaline lysis for industrial scale cGMP production of pharmaceutical grade plasmid-DNA. AB - Plasmid DNA for biopharmaceutical applications is mainly produced in E. coli cells. The first and most crucial step for recovering the plasmid is the cell lysis. Governed by the physico-chemical properties of the polynucleotide, alkaline lysis has been the lysis-method of choice. This chemical disintegration technique was initially developed for the lab scale and non-pharmaceutical applications. A continuous, fully automated and closed system combining alkaline lysis, neutralization and clarification in one gentle and generic operation was developed. This system consists of a three units. One unit controls mixing and contact time during the alkaline treatment, another one controls the neutralization and the concurrent formation of flocs and a third one the separation of flocs and pDNA containing lysate. Based on optimization experiments the selected process parameters resulted in yields up to 100% and homogeneities comparable to that obtained by gentle manual lysis. The process does not need enzymes and it is scalable and routinely used for cGMP-production of pharmaceutical grade plasmid DNA from 200 L fermentations. PMID- 17129626 TI - Delta and theta oscillations as risk markers in adolescent offspring of alcoholics. AB - BACKGROUND: Visual P300 is consistently lower in alcohol-dependent individuals, their offspring and subjects at risk. Delta and theta event-related oscillations (ERO) are the major contributors to the P300 signal. The total and evoked power in delta and theta bands in the 300 to 700 ms post-stimulus window (corresponding to the zone of P300 maxima) was compared between adolescent offspring of alcoholics (high-risk) and age-matched normal controls (low-risk), to assess the utility of the risk markers. METHODS: EEG was recorded during the performance of a visual oddball task. The S-transform algorithm decomposed the EEG signals into different frequency bands and the group differences in total and evoked power in the oscillatory responses during the P300 time window (300 to 700 ms) were analyzed using a multivariate design. Similar analysis was performed on P300 peak amplitude for the target. RESULTS: The high-risk group showed significantly lower parietal post-stimulus evoked and total power in the delta band for targets. A decrease in total power was seen centrally and parietally in the theta band. The P300 peak amplitude in the parietal electrodes was also significantly lower in the high-risk group. CONCLUSIONS: The decreased total theta power and total and evoked delta power for visual targets in high risk individuals may serve as an endophenotypic marker in the development of alcoholism and other disinhibitory disorders. The differences seen between the offspring of alcoholics and controls may have a cholinergic basis. The ERO measures appear to be more robust than the P300 amplitude in differentiating the groups. PMID- 17129628 TI - Characterization of a bifunctional aminoacylase/carboxypeptidase from radioresistant bacterium Deinococcus radiodurans R1. AB - The gene encoding a Deinococcus radiodurans R1 bifunctional aminoacylase/carboxypeptidase (DR_ACY/CP) was amplified by polymerase chain reaction and cloned into pQE-30 to generate pQE-DRAC. The cloned gene consists of an open reading frame of 1197 bp encoding a protein with a molecular mass of 42,729 Da. The predicted amino acid sequence shows high homology with those of Geobacillus kaustophilus aminoacylase, Geobacillus stearothermophilus aminoacylase, Pyrococcus horikoshii carboxypeptidase/aminoacylase and Thermoanaerobacter tengcongensis aminoacylase/carboxypeptidase. The expressed enzyme was purified from the crude extract of IPTG-induced Escherichia coli M15 (pQE-DRAC) to homogeneity by nickel-chelate chromatography. The molecular mass of the purified enzyme was determined to be 43kDa by SDS-PAGE. Maximal aminoacylase activity with N-acetyl-methionine as the substrate occurred at pH 8.0 and 40 degrees C in the sodium phosphate buffer. The aminoacylase activity was strongly inhibited by metal-chelating agents, and was largely restored by divalent cations, such as Co(2+), Mn(2+) and Ni(2+). The purified enzyme had broad specificity toward N-acetylated L-amino acids as well as N-CBZ-peptides. Carboxypeptidase activity of DR_ACY/CP to N-CBZ-Gly-Ala exhibited K(m) and k(cat) values of 4.3mM and 28s(-1), respectively. The enzyme also had activity toward the cell wall-related substrates, D-Ala-Gly, D-Ala-Gly-Gly and L-Orn-L-Ala. PMID- 17129629 TI - Role of envelope processing and gp41 membrane spanning domain in the formation of human immunodeficiency virus type 1 (HIV-1) fusion-competent envelope glycoprotein complex. AB - Human immunodeficiency virus type 1 (HIV-1) entry into target cells is directed by the envelope (Env) glycoproteins, which are present on the surface of HIV-1 virion or infected cells in the form of trimers consisting of gp120/gp41 complexes. The surface subunit, gp120, initiates the entry process by interacting sequentially with the CD4 receptor and a co-receptor, thereby inducing a conformational change that allows the transmembrane (TM) gp41 subunit to mediate fusion between viral and target cell membranes. Cleavage of Env into its gp120 and gp41 components is necessary for activation of its fusogenic activity. Here, the gp41 TM glycoprotein was altered by either deleting an isoleucine residue (DeltaI642) in a critical region of its ectodomain or by substituting its membrane spanning domain (MSD) by that of the influenza hemagglutinin (HA) glycoprotein (TM-HA) to examine the contribution of these regions to Env functions. Characterization of these mutant forms of gp41 revealed that they both affected the infectivity of pseudotyped virions, however, through distinct defects in Env functions. While deletion of Ile 642 drastically altered processing of Env, replacement of gp41 MSD by that of HA led to a marked fusion defect even though the TM-HA Env was efficiently processed and incorporated into viral particles. Interestingly, both DeltaI642 and TM-HA Env were found to act as trans dominant-negative mutant of viral infectivity, presumably via their ability to form hetero-oligomers with wild type Env. Together, these results support a previously proposed model whereby all three gp120-gp41 monomers must be cleaved for the Env homo-trimer to function and suggest that the gp41 MSD plays a critical role in the formation of fusion-competent Env trimers. PMID- 17129630 TI - Molecular analysis of double-stranded RNAs reveals complex infection of grapevines with multiple viruses. AB - The table grape variety "Waltham Cross" was infected with Leafroll and Shiraz Disease. To reveal specific viruses that are associated with the diseased plants, we used an RT-PCR-based strategy to determine partial genome sequences of these viruses. Upon cloning and sequencing of the RT-PCR products, we detected seven groups of viral variants that are related to four species of the Closteroviridae: Grapevine leafroll-associated virus 1, 2, 3, and 5, in addition to Rupestris stem pitting-associated virus. The population composition of GLRaV-2 and GLRaV-3-like viruses is complex and consists of two or three distinct groups of viral variants. Based on the consensus sequence of several GLRaV-2 strains, we designed a pair of broad-spectrum primers (GLR2-4 and GLR2-5) and used them to detect a range of GLRaV-2 variants from "Waltham Cross". Moreover, we identified a novel group of viral variants from the diseased grapevines, which possess a stretch of 19 nucleotides inserted in the 3' non-coding region as compared to strain "PN" and "93/955" for which the complete genomes have been sequenced. In contrast, the population composition of GLRaV-1 and GLRaV-5-like virus seems to be more uniform and each consists of a single viral variant. Furthermore, the central 5.7kb genomic region encompassing ORF1b-ORF4 of the GLRaV-1 isolate detected in "Waltham Cross" was sequenced. The new isolate is designated GLRaV-1 "WC", which differs from GLRaV-1 "Type" by 16% in nucleotide sequence. The taxonomic standing of the GLRaV-5-like and GLRaV-3-like viruses detected in "Waltham Cross" is discussed. PMID- 17129631 TI - A molecular epidemiological study targeting the glycoprotein gene of rabies virus isolates from China. AB - A group of 31 rabies viruses (RABVs), recovered primarily from dogs, one deer and one human case, were collected from various areas in China between 1989 and 2006. Complete G gene sequences determined for these isolates indicated identities of nucleotide and amino acid sequences of >or=87% and 93.8%, respectively. Phylogenetic analysis of these and some additional Chinese isolates clearly supported the placement of all Chinese viruses in Lyssavirus genotype 1 and divided all Chinese isolates between four distinct groups (I-IV). Several variants identified within the most commonly encountered group I were distributed according to their geographical origins. A comparison of representative Chinese viruses with other isolates retrieved world-wide indicated a close evolutionary relationship between China group I and II viruses and those of Indonesia while China group III viruses formed an outlying branch to variants from Malaysia and Thailand. China group IV viruses were closely related to several vaccine strains. The predicted glycoprotein sequences of these RABVs variants are presented and discussed with respect to the utility of the anti-rabies biologicals currently employed in China. PMID- 17129632 TI - Investigating the mechanism of enhanced cytotoxicity of HPMA copolymer-Dox-AGM in breast cancer cells. AB - Recently we have described an HPMA copolymer conjugate carrying both the aromatase inhibitor aminoglutethimide (AGM) and doxorubicin (Dox) as combination therapy. This showed markedly enhanced in vitro cytotoxicity compared to the HPMA copolymer-Dox (FCE28068), a conjugate that demonstrated activity in chemotherapy refractory breast cancer patients during early clinical trials. To better understand the superior activity of HPMA copolymer-Dox-AGM, here experiments were undertaken using MCF-7 and MCF-7ca (aromatase-transfected) breast cancer cell lines to: further probe the synergistic cytotoxic effects of AGM and Dox in free and conjugated form; to compare the endocytic properties of HPMA copolymer-Dox AGM and HPMA copolymer-Dox (binding, rate and mechanism of cellular uptake); the rate of drug liberation by lysosomal thiol-dependant proteases (i.e. conjugate activation), and also, using immunocytochemistry, to compare their molecular mechanism of action. It was clearly shown that attachment of both drugs to the same polymer backbone was a requirement for enhanced cytotoxicity. FACS studies indicated both conjugates have a similar pattern of cell binding and endocytic uptake (at least partially via a cholesterol-dependent pathway), however, the pattern of enzyme-mediated drug liberation was distinctly different. Dox release from PK1 was linear with time, whereas the release of both Dox and AGM from HPMA copolymer-Dox-AGM was not, and the initial rate of AGM release was much faster than that seen for the anthracycline. Immunocytochemistry showed that both conjugates decreased the expression of ki67. However, this effect was more marked for HPMA copolymer-Dox-AGM and, moreover, only this conjugate decreased the expression of the anti-apoptotic protein bcl-2. In conclusion, the superior in vitro activity of HPMA copolymer-Dox-AGM cannot be attributed to differences in endocytic uptake, and it seems likely that the synergistic effect of Dox and AGM is due to the kinetics of intracellular drug liberation which leads to enhanced activity. PMID- 17129633 TI - VDR polymorphisms influence the immune response in type 1 diabetic children from Santiago, Chile. AB - OBJECTIVE: To evaluate the influence of ApaI, BsmI and TaqI polymorphisms of the VDR gene and HLA-DQB1* alleles in type 1 diabetic children and to assess their possible relationship with circulating levels of 25-hydroxyvitamin D(3), auto antibodies, and INFgamma/TGFbeta1 cytokines levels in Chilean cases and controls. METHODS: DNA and serum samples from 216 newly diagnosed type 1 diabetic and 203 unrelated control children were evaluated for IA-2 and GAD(65) auto-antibodies, 25-hydroxyvitamin D(3) levels, HLA-DQB1* alleles, and VDR gene polymorphisms. RESULTS: The frequency of the b allele and the bb genotype in type 1 diabetic patients was significantly lower compared with the control group (0.635 versus 0.749, p<0.01 and 0.370 versus 0.567, p<0.04). 25-Hydroxyvitamin D(3) levels showed no differences between type 1 diabetic and healthy children. In cases, 25 hydroxyvitamin D(3) levels were not associated with a special auto-antibodies profile according to the presence or absence of GAD(65)(+) or IA-2(+). The haplotype combination BAT was higher in cases (0.062 versus 0.019, p<0.0022) and bAT was more frequent in controls (0.266 versus 0.180, p<0.003). In cases, the aaBbTT genotype showed the most significant increase in TGFbeta1 level across the VDR categories. Finally, when considering the HLA class II risk genotype (DQB1*0302) and the VDR genotypes (AabbTT and aabbTT), higher levels of GAD(65), IA-2 and TGFbeta1 were observed among diabetic children. CONCLUSION: We found an association between a VDR polymorphism (BsmI) and type 1 diabetes. An association was found of AabbTT and aabbTT genotypes and the HLA-DQB1*0302 allele with high levels of GAD(65), IA-2 and TGFbeta1. PMID- 17129634 TI - Irradiated volume and the risk of fatal hemoptysis in patients submitted to high dose-rate endobronchial brachytherapy. AB - To determine risk factors associated with fatal hemoptysis (FH) in endobronchial high dose-rate brachytherapy (EHDRB) 84 patients treated with EHDRB from January 1991 to June 2002 were studied. Clinical and technical parameters (including treatment volumes) were analyzed. Eight (9.5%) patients died of FH, all but one with recurrent or persistent local disease. Median interval until death due to FH was 4 months versus 6 months for the whole group. The only factor with significant correlation with FH was the 100% isodose volume (V100) (p=0.04). Larger irradiated volumes were related to FH. Analysis of volume parameters is suggested, together with the dose and number of fractions prescribed for each patient. PMID- 17129635 TI - Repeat 18F-FDG PET for monitoring neoadjuvant chemotherapy in patients with stage III non-small cell lung cancer. AB - PURPOSE: The relevance of (18)F-FDG PET for staging non-small cell lung cancer (NSCLC), in particular for the detection of lymph node or distant metastases, has been shown in several studies. The value of FDG-PET for therapy monitoring in NSCLC, in contrast, has not yet been sufficiently analysed. Aim of this study was to evaluate FDG-PET for monitoring treatment response during and after neoadjuvant radiochemotherapy (NARCT) in advanced NSCLC. METHODS: Sixty-five patients with histologically proven NSCLC stage III initially underwent three FDG PET investigations, during NARCT prior to initiating radiation, and post-NARCT. Changes of FDG-uptake in the primary tumour at two time-points during NARCT were analysed concerning their impact on long-term survival. RESULTS: The mean maximum FDG uptake (standardized uptake value, SUVmax) of the whole group decreased significantly during NARCT (SUVmax PET 1: 14.9+/-4.0, SUVmax PET 3: 5.5+/-2.4, p=0.004). The difference between initial FDG uptake (PET 1) and uptake after induction chemotherapy (PET 2) was found to be highly predictive for long-term survival patients which had a greater than 60% decreases in their SUV change had a significantly longer survival than those below this threshold (5-year-survival 60% versus 15%, p=0.0007). Patients who had a lower than 25% decrease in their SUV change had a 5-years-survival lower than 5%. Furthermore, the difference between initial FDG uptake (PET 1) and uptake after completion of the whole NARCT (PET 3) was predictive for survival when 75% was applied as cut-off (p=0.02). However, the level of significance was considerably lower. CONCLUSION: FDG-PET is suitable for therapy monitoring in patients with stage III NSCLC. The decrease of FDG uptake during induction chemotherapy is highly predictive for patient outcome. PMID- 17129636 TI - Cryptic species as a window on diversity and conservation. AB - The taxonomic challenge posed by cryptic species (two or more distinct species classified as a single species) has been recognized for nearly 300 years, but the advent of relatively inexpensive and rapid DNA sequencing has given biologists a new tool for detecting and differentiating morphologically similar species. Here, we synthesize the literature on cryptic and sibling species and discuss trends in their discovery. However, a lack of systematic studies leaves many questions open, such as whether cryptic species are more common in particular habitats, latitudes or taxonomic groups. The discovery of cryptic species is likely to be non-random with regard to taxon and biome and, hence, could have profound implications for evolutionary theory, biogeography and conservation planning. PMID- 17129637 TI - Bacisubin, an antifungal protein with ribonuclease and hemagglutinating activities from Bacillus subtilis strain B-916. AB - An antifungal protein, with a molecular mass of 41.9 kDa, and designated as bacisubin, was isolated from a culture of Bacillus subtilis strain B-916. The isolation procedure consisted of ion exchange chromatography on DEAE-Sepharose Fast Flow, and fast protein liquid chromatography on Phenyl Sepharose 6 Fast Flow and hydroxyapatite columns. The protein was adsorbed on all three chromatographic media. Bacisubin exhibited inhibitory activity on mycelial growth in Magnaporthe grisease, Sclerotinia sclerotiorum, Rhizoctonia solani, Alternaria oleracea, A. brassicae and Botrytis cinerea. The IC50 values of its antifungal activity toward the last four fungal species were 4.01 microM, 0.087 microM, 0.055 microM and 2.74 microM, respectively. Bacisubin demonstrated neither protease activity, nor protease inhibitory activity. However, it manifested ribonuclease and hemagglutinating activities. PMID- 17129638 TI - Evidence for a new angiotensin-(1-7) receptor subtype in the aorta of Sprague Dawley rats. AB - We have recently described, in the mouse aorta, the vasodilator effect of angiotensin-(1-7) (Ang-(1-7)) was mediated by activation of the Mas Ang-(1-7) receptor and that A-779 and D-Pro7-Ang-(1-7) act as Mas receptor antagonists. In this work we show pharmacological evidence for the existence of a different Ang (1-7) receptor subtype mediating the vasodilator effect of Ang-(1-7) in the aorta from Sprague-Dawley (SD) rats. Ang-(1-7) induced an endothelium-dependent vasodilator effect in aortic rings from SD rats which was inhibited by removal of the endothelium and by L-NAME (100 microM) but not by indomethacin (10 microM). The Ang-(1-7) receptor antagonist D-Pro7-Ang-(1-7) (0.1 microM) abolished the vasodilator effect of the peptide. However, the other specific Ang-(1-7) receptor antagonist, A-779 in concentrations up to 10 microM, did not affect vasodilation induced by Ang-(1-7). The Ang II AT1 and AT2 receptors antagonists CV11974 (0.01 microM) and PD123319 (1 microM), respectively, the bradykinin B2 receptor antagonist HOE 140 (1 microM) and the inhibitor of ACE captopril (10 microM) did not change the effect of Ang-(1-7). Our results show that in the aorta of SD rats, the vasodilator effect of Ang-(1-7) is dependent on endothelium-derived nitric oxide. This effect is mediated by the activation of Ang-(1-7) receptors sensitive to D-Pro7-Ang-(1-7), but not to A-779, which suggests the existence of a different Ang-(1-7) receptor subtype. PMID- 17129639 TI - Intravenous CCK-8, but not GLP-1, suppresses ghrelin and stimulates PYY release in healthy men. AB - We have investigated the effects of exogenous CCK-8 and GLP-1, alone and in combination, on ghrelin and PYY secretion. Nine healthy males were studied on four occasions. Plasma ghrelin and PYY concentrations were measured during 150 min intravenous infusions of: (i) isotonic saline, (ii) CCK-8 at 1.8 pmol/kg/min, (iii) GLP-1 at 0.9 pmol/kg/min or (iv) CCK-8 and GLP-1 combined. CCK-8 markedly suppressed ghrelin and stimulated PYY when compared with control between t=0-120 min (P<0.001 for both). GLP-1 had no effect on ghrelin, but decreased PYY slightly at 120 min (P<0.05). During infusion of CCK-8+GLP-1, there was comparable suppression of ghrelin (P<0.001), but the stimulation of PYY was less (P<0.001), than that induced by CCK-8, between t=20-120 min. In conclusion, in healthy subjects, in the doses evaluated, exogenous CCK-8 suppresses ghrelin and stimulates PYY, and exogenous GLP-1 has no effect on ghrelin and attenuates the effect of CCK-8 on PYY. PMID- 17129640 TI - Resetting of central and peripheral circadian oscillators in aged rats. AB - The mammalian circadian timing system is affected by aging. Analysis of the suprachiasmatic nucleus (SCN) and of other circadian oscillators reveals age related changes which are most profound in extra-SCN tissues. Some extra-SCN oscillators appear to stop oscillating in vivo or display altered phase relationships. To determine whether the dynamic behavior of circadian oscillators is also affected by aging we studied the resetting behavior of the Period1 transcriptional rhythm of peripheral and central oscillators in response to a 6h advance or delay in the light schedule. We employed a transgenic rat with a luciferase reporter to allow for real-time measurements of transcriptional rhythmicity. While phase resetting in the SCN following an advance or a delay of the light cycle appears nearly normal in 2-year-old rats, resynchronization of the liver was seriously disrupted. In addition, the arcuate nucleus and pineal gland exhibited faster resetting in aged rats relative to 4-8-month-old controls. The consequences of these deficits are unknown, but may contribute to organ and brain diseases in the aged as well as the health problems that are common in older shift-workers. PMID- 17129641 TI - Synthesis, antimicrobial, and anti-inflammatory activities of novel 2-(1 adamantyl)-5-substituted-1,3,4-oxadiazoles and 2-(1-adamantylamino)-5-substituted 1,3,4-thiadiazoles. AB - Reaction of 1-adamantanecarbonyl chloride with certain carboxylic acid hydrazides in pyridine yielded the corresponding N-acyl adamantane-1-carbohydrazide derivatives 3a-j, which were cyclized to the corresponding 2-(1-adamantyl)-5 substituted-1,3,4-oxadiazoles 4a-j via heating with phosphorus oxychloride. Treatment of 1-adamantylisothiocyanate with some carboxylic acid hydrazides in ethanol yielded the corresponding 1-acyl-4-(1-adamantyl)-3-thiosemicarbazides 7a g, which were cyclized to the corresponding 2-(1-adamantylamino)-5-substituted 1,3,4-thiadiazole derivatives 8a-g. Compounds 4a-j, 7a-g, and 8a-g were tested for in vitro activities against a panel of Gram-positive and Gram-negative bacteria and the yeast-like pathogenic fungus Candida albicans. Several derivatives produced good or moderate activities particularly against the tested Gram-positive bacteria Bacillus subtilis. Meanwhile, compounds 4i and 8g displayed marked antifungal activity against C. albicans. In addition, the in vivo anti-inflammatory activity of the synthesized compounds was determined using the carrageenin-induced paw oedema method in rats. The oxadiazole derivatives 4c, 4g, 4i and 4j produced good dose-dependent anti-inflammatory activity. PMID- 17129642 TI - A survey of knowledge, attitudes, and behavior of Italian dentists toward immunization. AB - This study assessed the knowledge, attitude, and compliance to immunization guidelines among dental health-care professionals in Italy. A questionnaire was mailed to a random sample of 1000 dentists on demographic and professional characteristics; knowledge regarding infectious diseases that can be acquired/transmitted by the dentist and the vaccinations recommended; attitudes regarding the vaccinations; self-assessed information about previous vaccinations. A total of 369 dentists responded. Only 44.1 and 32.4% correctly indicated all infections that can be acquired or transmitted during their activity. Half of the dentists knew that they should be vaccinated against hepatitis B and influenza and this knowledge was significantly higher in those older, who correctly indicated that hepatitis B and influenza can be acquired and transmitted during their activity, and in those with a positive attitude towards vaccinations. Those younger, involved in specialties with low blood contact, and who did not know that hepatitis B and influenza are vaccinations recommended, were more likely to believe that is important for them to receive vaccinations. A large proportion (85.7%) reported receiving the hepatitis B vaccine, but only 56.2% the three doses. Those with a lower number of years of activity, those who knew that hepatitis B can be acquired by the dentist, those who did not need information, and those who were informed from guidelines, educational courses, and dental associations were more likely to have received the three doses or to be natural immunizated. Training and educational interventions are needed to improve knowledge and immunization coverage. PMID- 17129643 TI - The experience of Middle Eastern men living in Sweden of maternal and child health care and fatherhood: focus-group discussions and content analysis. AB - OBJECTIVE: To describe how men from the Middle East experience Swedish maternity and child health care. An integral part of the aim of this study has also been to describe the experiences of men from the Middle East when becoming and being a father in Sweden. DESIGN, SETTING AND PARTICIPANTS: an exploratory, qualitative study using focus-group discussions and individual interviews, with a semi structured interview guide and content analysis. A total of 16 men participated. Ten Arabic-speaking men from the Middle East living in Sweden participated in three focus-group discussions. Six men from the Middle East living in Sweden, and speaking Swedish, participated in individual interviews. FINDINGS: three main categories were developed: meeting empathic professionals; finding new positions within the family; and experiencing social demands. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: seeing their partners being met individually and with empathy by midwives and child health-care nurses encouraged men to become involved in areas not previously open to them (i.e. pregnancy, childbirth and the care of babies and young children). As the women often lacked knowledge of Swedish, they depended on the help of their partners when meeting maternity and child health-care professionals. The men found the experience of living in an alien country difficult. They were often unemployed, felt they were a burden to their wives after emigrating to Sweden, and that they were no longer a suitable role model for their children. PMID- 17129644 TI - Open reduction of the radial head with ulnar osteotomy and annular ligament reconstruction for bilateral congenital radial head dislocation: a case with long term follow-up. AB - We report the 9 year follow-up results of treatment of a 5 year-old boy with bilateral congenital radial head dislocation by open reduction of the radial head and ulnar osteotomy with annular ligament reconstruction and discuss the management of this condition. PMID- 17129645 TI - Extensor tendon entrapment in volarly displaced distal radial fracture. PMID- 17129646 TI - Immediate mobilisation following corrective osteotomy of distal radius malunions with cancellous graft and volar fixed angle plates. AB - The purpose of the study was to examine the reliability with which a specific technique of corrective osteotomy of malunions of the distal radius combined with early mobilisation could both restore the normal anatomic parameters of the radius and achieve a functional range of motion with good strength. Corrective osteotomy of the distal radius was performed through a volar approach using a fixed angle volar plate and cancellous bone graft from the iliac crest in 19 patients of mean age 50 years with initial malunions with a mean dorsal tilt of 36 degrees and 7 mm of ulnar variance. An immediate mobilisation programme was started. All healed at a mean of less than 12 weeks (including two heavy smoking patients who required repeat cancellous bone grafting to achieve final union) to achieve a total arc of wrist motion around 120 degrees, forearm rotation of 158 degrees and grip strength which was 80% of contralateral. This treatment strategy was judged to be straightforward and effective. PMID- 17129648 TI - Effects of plant arsenic uptake and heavy metals on arsenic distribution in an arsenic-contaminated soil. AB - This study examined the effects of heavy metals and plant arsenic uptake on soil arsenic distribution. Chemical fractionation of an arsenic-contaminated soil spiked with 50 or 200mg kg(-1) Ni, Zn, Cd or Pb was performed before and after growing the arsenic hyperaccumulator Pteris vittata L for 8weeks using NH(4)Cl (water-soluble plus exchangeable, WE-As), NH(4)F (Al-As), NaOH (Fe-As), and H(2)SO(4) (Ca-As). Arsenic in the soil was present primarily as the recalcitrant forms with Ca-As being the dominant fraction (45%). Arsenic taken up by P. vittata was from all fractions though Ca-As contributed the most (51-71% reduction). After 8weeks of plant growth, the Al-As and Fe-As fractions were significantly (p<0.01) greater in the metal-spiked soils than the control, with changes in the WE-As fraction being significantly (p=0.007) correlated with plant arsenic removal. The plant's ability to solubilize soil arsenic from recalcitrant fractions may have enhanced its ability to hyperaccumulate arsenic. PMID- 17129649 TI - The use of sedimentary %C, %N, delta(15)N, and Pb concentrations to assess historical changes in anthropogenic influence on Portuguese estuaries. AB - Vertical profiles of C, N, delta(15)N and Pb were measured in the Mondego and Mira estuaries as markers that conveyed notions as to the relative influence of anthropogenic influence over the past decades. Recent carbon changes in both estuaries may reflect changes in estuarine productivity, probably as a consequence of sediment reworking and erosion, and also of losses in salt marsh area and dwarf eelgrass beds. delta(15)N values began to diverge considerably before the %C and %N and were higher in Mondego. delta(15)N signatures detected N enrichment at relatively low rates, and indicated that Mondego received more enriched N than Mira. Lead concentrations differed between estuaries, with higher concentrations in Mondego. The secular increase in %N, Pb, and delta(15)N signatures was significantly related to human density in the watersheds of the estuaries and were sensitive indicators of anthropogenic activity. PMID- 17129650 TI - XXII IUFRO World Congress, 2005 Brisbane - air pollution and climate change: a global overview of the effects on forest vegetation. PMID- 17129651 TI - Psychological distress among parents of children with mental retardation in the United Arab Emirates. AB - This study was designed to identify predictors of parental stress and psychological distress among parents of children with mental retardation in the United Arab Emirates. It examined the relative contributions of child characteristics, parents' sociodemographics, and family environment to parental stress and psychological distress. Participants were parents of 225 mentally retarded children, of whom 113 were fathers and 112 were mothers. Measures of parental stress (QRS-F), psychiatric symptom index (PSI) and family environment scale (FES) were administered in an interview format. Hierarchical multiple regression was used to predict parental stress and psychological distress. The results indicate that the model containing all three predictor blocks, child characteristics, parents' sociodemographics, and family environment, accounted for 36.3% and 22.5% of parental stress and parents' psychiatric symptomatology variance, respectively. The age of the child was significantly associated with parents' feelings of distress and psychiatric symptom status, and parental stress was less when the child was older. Parents reported more psychiatric symptomatology when the child showed a high level of dysfunction. Fathers' work appeared to be a significant predictor of parental stress, indicating that for fathers who were not working the level of stress was higher than fathers who were working. Lower socioeconomic level was associated with greater symptom rates of cognitive disturbance, depression, anxiety, and despair among parents. Among the family environment variables, only the personal growth dimension stood out as a predictor of parental stress. An orientation toward recreational and religious pursuits, high independence, and intellectual and recreational orientations were associated with lower levels of parental stress. On the other hand, parents in achievement-oriented families showed elevated levels of parental stress. Implications for prevention, and intervention as well as parent training and system-oriented counseling programs are discussed. PMID- 17129652 TI - Is attachment style a source of resilience against health inequalities at work? AB - The argument that 'indirect selection' is a contributory factor to health inequality has included ideas about personal characteristics that may originate in childhood and increase the likelihood of both poor health and disadvantaged social position in adulthood. The concept of protective resilience makes a similar but converse argument: that positive characteristics acquired at one phase of life may enable individuals to withstand later adversity. The increasing richness of data from longitudinal studies now allows us to examine these processes more closely over a longer period of life. In this paper we show that attachment style, a psychological characteristic thought to be associated with the style of parenting encountered during early childhood, may act as a source of resilience in the face of educational disadvantage. Men in mid-life who were not burdened with anxious or avoidant attachment styles seem to have been more likely to overcome the disadvantage of a lower level of educational attainment and progress up the ladder of Civil Service grades in the English Whitehall II study. As it is not strongly related to parents' social class, it can be argued that attachment style has acted as a source of upward social mobility which is also likely to reinforce better health in later life. PMID- 17129653 TI - Manipulating noise frequencies alters hemispheric contributions to decision making. AB - Participants listened to the Asian disease problem framed in terms of either gains or losses and chose between two plans to combat the disease. All participants heard the problem embedded in other sounds; for some it was the relatively lower-frequency information, and for others it was the relatively higher-frequency information. The classic framing effect appeared only for those participants for whom the problem was the relatively lower-frequency information (p<.05). These results suggest that mixing filtered speech signals and noise may be a way to assess the role of the left and right hemisphere in various aspects of decision making. PMID- 17129654 TI - Improvement in antipsychotic-related metabolic disturbances in patients with schizophrenia switched to ziprasidone. AB - PURPOSE: This study evaluated changes in weight, glucose and lipid metabolism in patients with schizophrenia and antipsychotic-related metabolic disturbances who were switched to ziprasidone. METHODS: Eighty-four outpatients with schizophrenia or schizoaffective disorder also having glucose intolerance, diabetes, dyslipidemia or weight gain related to their antipsychotic treatment were switched to ziprasidone. Clinical status was assessed using the Clinical Global Impression of Severity (CGI-S) and Improvement (CGI-I) scales and the Positive and Negative Syndrome Scale (PANSS). Assessment scales, weight, glucose and lipids were measured at baseline and at three and six months of ziprasidone treatment. RESULTS: Significant baseline to endpoint reductions were seen in mean weight (-5.1 kg), Body Mass Index (BMI; -1.6 kg/m(2)), serum glucose (-14.0 mg/dL), total cholesterol (-24.1 mg/dL), and triglyceride leves (-46.2 mg/dL). Mean PANSS total score improved 13.9% after 6 months of treatment with ziprasidone. A proportion (34.3%) of patients were classified as much improved in the CGI-I. CONCLUSIONS: Switching patients with schizophrenia to ziprasidone when metabolic disturbances are detected may improve these side effects and result in an improved overall outcome. PMID- 17129655 TI - Hemoglobin adducts of the human bladder carcinogen o-toluidine after treatment with the local anesthetic prilocaine. AB - Prilocaine, a widely used local anesthetic, is metabolized to o-toluidine which is classified as human carcinogen. We aimed to assess the impact of prilocaine treatment on hemoglobin adducts from o-toluidine. Blood samples were obtained before and 24h after receiving prilocaine local anesthesia (Xylonest, 100mg) from 20 head and neck surgery patients and 6 healthy volunteers. Hemoglobin adducts of o-toluidine and 4-aminobiphenyl were determined by gas chromatography/mass spectrometry. Hemoglobin adducts of o-toluidine were significantly increased 24h after 100mg prilocaine-treatment by 21.6+/-12.8ng/g hemoglobin (mean+/-S.D., N=26; P<0.0001). This corresponds to a 6-360-fold increase of o-toluidine adduct levels in 25 patients from 0.54+/-0.95ng/g before treatment to 22.0+/-13.2ng/g 24h after surgery (mean+/-S.D.). Because of an extremely high background level the increase was only 1.6-fold in one patient (40.9ng/g before and 64.4ng/g 24h after prilocaine injection). Current smoking had no influence on background values and on the increase of o-toluidine adducts. No treatment-related differences were seen in mean hemoglobin adduct levels of 4-aminobiphenyl which were significantly higher in smokers, 0.149+/-0.096ng/g (mean+/-S.D., N=8) as compared to nonsmokers 0.036+/-0.035ng/g (mean+/-S.D., N=16; P<0.01). In conclusion, prilocaine anesthesia leads to a massive increase of hemoglobin adducts of the carcinogenic arylamine o-toluidine. This implies a carcinogenic risk which should be taken into account in preventive hazard minimization. PMID- 17129656 TI - Asymmetric fluorogenic organophosphates for the development of active organophosphate hydrolases with reversed stereoselectivity. AB - In order to enhance the enzymatic detoxification rate of organophosphorus (OP) nerve agents we have searched for more active variants of recombinant mammalian paraoxonase (PON1). We have previously identified three key positions in PON1 that affect OP hydrolysis: Leu69, Val346 and His115, that significantly enhance the hydrolysis of cyclosarin (GF), soman, chlorpyrifos-oxon (ChPo), O-isopropyl-O (p-nitrophenyl)methylphosphonate (IMP-pNP) and diisopropyl fluorophosphate (DFP). GC/FPD analysis compared to residual AChE inhibition assay displayed stereoselective hydrolysis of GF, soman and IMP-pNP, indicating that wild type PON1 and its variant V346A are more active toward the less toxic P(+) optical isomer. In order to obtain new PON1 variants with reversed stereoselectivity, displaying augmented activity toward the more toxic isomer P(-) of nerve agents, we synthesized new asymmetric fluorogenic OPs (Flu-OPs). Six Flu-OPs were prepared containing either ethyl (E), cyclohexyl (C) or pinacolyl (P) alkyl radicals attached to methyl-phosphonyl (MP) moiety analogous to the structure of VX, GF and soman, respectively. The fluorescent moieties are either 3-cyano-4 methyl-7-hydroxy coumarin (MeCyC) or 1,3-dichloro-7-hydroxy-9,9-dimethyl-9H acridin-2-one (DDAO). The kinetics of AChE and BChE inhibition by these new Flu OPs display k(i) values 8.5x10(4) to 8.5x10(7) and 5x10(4) to 2x10(6)M(-1)min( 1), respectively. EMP-MeCyC and EMP-DDAO are the most active inhibitors of AChE whereas CMP-MeCyC and CMP-DDAO are better inhibitors of BChE than AChE, indicating accommodation of bulky cyclohexyl group inside the active site of BChE. PMP-MeCyC and PMP-DDAO are the least active inhibitors of both AChE and BChE. CMP-MeCyC and CMP-DDAO were significantly detoxified only by the five-site mutations PON1 variant L69V/S138L/S193P/N287D/V346A. Degradation kinetics of Flu OPs measured by increase in absorbance of the released fluorogenic group was fit by a two exponential function, indicating faster hydrolysis of the less toxic optical isomer. Interestingly, wt PON1 caused only 50% degradation of racemic EMP MeCyC, CMP-MeCyC and CMP-DDAO indicating complete hydrolysis of P(+) isomer. This remarkable stereoselectivity was used for the enzymatic separation of the P(-) isomer of CMP-MeCyC. The bimolecular rate constant k(i) for human AChE inhibition by the isolated P(-) isomer of CMP-MeCyC is five-fold larger than that of its P(+) isomer. The marked preference of wt PON1 toward P(+) stereo-isomer of CMP MeCyC and CMP-DDAO renders their P(-) stereo-isomers suitable for the selection of new OP hydrolase variants with reversed stereoselectivity. PMID- 17129657 TI - The perinatal autopsy: pertinent issues in multicultural Western Europe. AB - Western Europe is in a demographic transition with increasing multicultural societies. Health professionals have to understand the background, religious and cultural aspects of parents to counsel them regarding an autopsy in the event of a perinatal loss. Autopsy rates have declined over the past decades, the major limiting factor being the granting of permission for an autopsy, possibly because of adverse publicity or reluctance of doctors to obtain consent. Autopsy has proved its value in revealing unsuspected findings. The public can be convinced of this utility by means of good information notwithstanding their religious or cultural background. PMID- 17129658 TI - Plasma uric acid levels do not correlate to plasma-evoked changes in endothelial function in women with preeclampsia. PMID- 17129659 TI - LAI inversion algorithm based on directional reflectance kernels. AB - Leaf area index (LAI) is an important ecological and environmental parameter. A new LAI algorithm is developed using the principles of ground LAI measurements based on canopy gap fraction. First, the relationship between LAI and gap fraction at various zenith angles is derived from the definition of LAI. Then, the directional gap fraction is acquired from a remote sensing bidirectional reflectance distribution function (BRDF) product. This acquisition is obtained by using a kernel driven model and a large-scale directional gap fraction algorithm. The algorithm has been applied to estimate a LAI distribution in China in mid July 2002. The ground data acquired from two field experiments in Changbai Mountain and Qilian Mountain were used to validate the algorithm. To resolve the scale discrepancy between high resolution ground observations and low resolution remote sensing data, two TM images with a resolution approaching the size of ground plots were used to relate the coarse resolution LAI map to ground measurements. First, an empirical relationship between the measured LAI and a vegetation index was established. Next, a high resolution LAI map was generated using the relationship. The LAI value of a low resolution pixel was calculated from the area-weighted sum of high resolution LAIs composing the low resolution pixel. The results of this comparison showed that the inversion algorithm has an accuracy of 82%. Factors that may influence the accuracy are also discussed in this paper. PMID- 17129660 TI - Observation and simulation of net primary productivity in Qilian Mountain, western China. AB - We modeled net primary productivity (NPP) at high spatial resolution using an advanced spaceborne thermal emission and reflection radiometer (ASTER) image of a Qilian Mountain study area using the boreal ecosystem productivity simulator (BEPS). Two key driving variables of the model, leaf area index (LAI) and land cover type, were derived from ASTER and moderate resolution imaging spectroradiometer (MODIS) data. Other spatially explicit inputs included daily meteorological data (radiation, precipitation, temperature, humidity), available soil water holding capacity (AWC), and forest biomass. NPP was estimated for coniferous forests and other land cover types in the study area. The result showed that NPP of coniferous forests in the study area was about 4.4 tCha(-1)y( 1). The correlation coefficient between the modeled NPP and ground measurements was 0.84, with a mean relative error of about 13.9%. PMID- 17129661 TI - Rangeland dynamics in southern Ethiopia: (1) botanical composition of grasses and soil characteristics in relation to land-use and distance from water in semi-arid Borana rangelands. AB - The distribution and compositional pattern of the grass layer, as well as soil quality of southern Ethiopian semi-arid rangelands were examined under three land use systems (communal land, government ranch and a traditional grazing reserve enclosure) and along a distance gradient (near, middle and far) from water sources. In total, 49 grass species were identified, 65% of which were perennials. Of the most commonly distributed highly desirable species, Chrysopogon aucheri showed the greatest frequency under the traditional grazing reserve (27.6%), and the lowest in the communal land (14.0%). The frequency of Cenchrus ciliaris was similar in all the study sites. The proportion of Lepthotrium senegalensis was low under the land-use systems (3.9%) and increased away from water sources (2.3%). Of the less desirable species, Sporobulus nervosus was extremely more abundant (<0.05) in the communal land (13.3%) than the traditional grazing reserve (3.3%) and the government ranch (1.9%). Sporobolus pyramidalis was dominant in the study sites, with similar (P>0.05) frequency percentages under the land-use systems and along the distance gradients from water. Basal cover was low and similar on all the study sites (3.3%). Concerning the soil texture, sand (71.1%) constituted the largest proportion, followed by silt (21.3%) and lastly, clay (7.7%). With regard to soil exchangeable cations, organic C and total nitrogen were generally low and did not show marked variation in all the study sites. The study showed significant differences for most of the vegetation variables between the communal land and the other land-uses. This suggested that grazing intensity was higher in the communal land and moderate in the government ranch and in the traditional grazing reserves (kallos). The lack of significant differences in most of the studied (vegetation) variables along the distance gradient from water could be ascribed to the fact that grazing disturbance has already exceeded a certain threshold of degradation. Under the present low states of soil nutrients and rainfall, cultivation is neither sustainable nor environmentally friendly and this will lead to further degradation of the soil in these marginal lands. PMID- 17129662 TI - Social and economic impacts of carbon sequestration and land use change on peasant households in rural China: a case study of Liping, Guizhou Province. AB - Numerous innovative approaches to mitigate effects of excessive emission of greenhouse gases (GHGs) on global climate change are being proposed and formulated. Sequestering carbon to terrestrial ecosystems represents one of the important clean development mechanisms. Reforestation through converting various non-forest lands to forests is undoubtedly an important dimension of carbon sequestration. Using Liping County in Guizhou Province as a case region, this study examines the perceived change in social and economic livelihoods of peasants and the factors responsible for the variations in the changes. The results of the study reveal that socio-economic changes associated with the government-financed project are multifaceted and profound. Because of the financial subsidies provided by the central government, this environmental action in many aspects can be regarded as a poverty reduction measure in the underdeveloped area where rural poverty is widespread. A majority of peasant households have benefited from project participation. The land conversion project with continued financial support also contributes to the social transformations of traditional rural society in remote areas to a more mobile, less subsistence agriculture-based, and open society. PMID- 17129663 TI - Property rights and grassland degradation: a study of the Xilingol pasture, Inner Mongolia, China. AB - The semi-private property rights arrangement called the Household Production Responsibility System (HPRS) was started in the early 1980s in Xilingol pasture of Inner Mongolia (China), and stimulated the development of stockbreeding. The grassland has been degrading severely with increasing numbers of livestock. Based on a historical review of property rights regimes in Inner Mongolia and empirical surveys in Xilingol pasture during 2001-2003, this paper assesses the implementation of HPRS and its impacts on incomes of households as well as the environmental impact on the grassland. It was found that HPRS does not mitigate the "Tragedy of the Commons", instead it has exacerbated the situation. It was also found that co-management of grassland and livestock among a few households presents a sustainable use of grassland to develop livestock breeding. We conclude with the recommendation that small-scale collective property rights systems should be encouraged in Xilingol pasture of Inner Mongolia. PMID- 17129664 TI - Do statistical inferences allowing three alternative decisions give better feedback for environmentally precautionary decision-making? AB - Environmental policies and guidelines often specify standards for environmental indicators. The first part of this paper argues that, where compliance with these standards is assessed with the help of statistical inference, an inference employing a three-alternatives decision rule can provide more sensible feedback to environmental managers for precautionary decision-making. The second part of the paper shows how a three-alternatives statistical inference about compliance with a percentile standard might be applied to a small number of observations using a non-parametric binomial interval. This interval expression of uncertainty results in the sample size requirements for various percentile ranks becoming explicit. PMID- 17129665 TI - Soil compaction--indicators for the assessment of harmful changes to the soil in the context of the German Federal Soil Protection Act. AB - Soil compaction in agriculture induced by large-scale equipment is of growing concern. Heavy wheel loads used in arable cropping have the potential to cause irreversible damage to the subsoil structure and may lead to harmful soil compaction. In order to sustain or improve soil health or fitness on a sustainable basis, indicators are needed to assess the changes in the soil structure and the respective soil functions. This requires an adequate verification of methods for distinguishing between the impairment of soil structure and the disruption of soil functions, and for the respective subject of protection. In this article the link between existing models of soil physical prognosis, practical guidelines, and criteria for the identification of affected soil structure is demonstrated, and a viable concept to distinguish harmful changes to the soil is presented. The concept consists of methods for soil physical analysis such as "pre-compression stress" and "loading ratio", practical recommendations for best management practice, and an indicator-based model for the identification of harmful subsoil compaction derived from a research project for the German Federal Environmental Agency, making it possible to determine for a respective location the required level of action far beyond the common practice of precautions against harmful soil compaction. PMID- 17129666 TI - Environmentally adjusted productivity measurement: an Australian case study. AB - This paper critically examines various methods for estimating productivity incorporating environmental effects for the Australian agricultural sector. The agricultural sector has been selected because of its strategic position in the economy of Australia. The findings of this study indicate that the application of environmentally adjusted productivity methods is a credible approach to measure productivity, in the context of sustainable development. Although the empirical findings of this research are case study specific, the results provide evidence supporting the adoption of these techniques to other sectors of the economy when measuring productivity and needing to be cognisant of sustainable development. The findings suggest that adjusting for the environmental impacts of soil erosion can result in higher or lower agricultural productivity depending on the assumptions we make regarding damage costs of erosion. It is argued in this paper that, for soil erosion in Australia, assumptions yielding higher productivity (i.e., upwardly adjusted) are justified. Finally, the findings of this study and the use of the methods presented point to important gaps in data availability. This gap needs to be addressed by policy makers if sustainable development objectives are to be credibly assessed using these techniques. PMID- 17129667 TI - Design and validation of a new screening instrument for lower urinary tract dysfunction: the bladder control self-assessment questionnaire (B-SAQ). AB - OBJECTIVES: To develop and validate a short patient self-assessment screening questionnaire: bladder control self-assessment questionnaire (B-SAQ) for the evaluation of lower urinary tract symptoms. This first validation study was undertaken amongst women. PATIENTS AND METHODS: Three hundred twenty-nine women attending general gynaecology and urogynaecology clinics completed both the B-SAQ and Kings Health questionnaire prior to medical consultation, and independent physician assessment of the presence of lower urinary tract symptoms (LUTS) and need for treatment. The psychometric properties of the B-SAQ were subsequently analysed. RESULTS: The B-SAQ was quick and easy to complete, with 89% of respondents completing all items correctly in less than 5 min. The internal consistency (Cronbach's alpha score 0.90-0.91), criterion validity (Pearson's correlation values of 0.79 and 0.81, p<0.0001 with the incontinence impact domain of the Kings Health questionnaire), and test-retest reliability of the questionnaire were good. The sensitivity and specificity of the questionnaire to identify patients with bothersome LUTS was 98% and 79%, respectively. CONCLUSIONS: LUTS are commonly underreported. Empowering patients to self-assess their bladder symptoms and the need for treatment will improve treatment-seeking behaviour. The B-SAQ is a psychometrically robust, short screening questionnaire that offers patients the ability to assess their bladder symptoms and the bother they cause, and the potential benefit of seeking medical help. PMID- 17129668 TI - STAT3 as a central mediator of neoplastic cellular transformation. AB - Much of the focus in understanding the molecular pathogenesis of tumors has centered on kinases that are activated in cancer. However, cancers driven by a diversity of activated kinases may have very similar pathological and clinical properties. This likely relates to the fact that the biological characteristics of a tumor are driven by the pattern of gene expression in that tumor, and that a wide spectrum of activating events at the cell surface and in the cytoplasm converge on a relatively small number of transcription factors that regulate the expression of key target genes. One transcription factor that has been found to be activated inappropriately in a wide range of human cancers is STAT3. STAT3 target genes are involved in fundamental events of tumor development including proliferation, survival, self-renewal, invasion, and angiogenesis. Furthermore, there is strong evidence that STAT3 is critical for these processes, in that inhibition of STAT3 by a variety of means can exert an anti-cancer effect. Since normal cells are relatively tolerant of interruption in STAT3 signaling, these findings suggest that STAT3 may also be an excellent target for the molecular therapy of cancer. PMID- 17129669 TI - Seasonal and spatial trends of suspended-particle associated polycyclic aromatic hydrocarbons in urban Shizuoka, Japan. AB - We characterized the monthly variations of 21 polycyclic aromatic hydrocarbons (PAHs) associated with suspended particles in the city of Shizuoka, Japan, over 12 months during 2001 and 2002. The fraction of fine particles (less than 2.5 microm diameter (PM2.5)) ranged from 40 to 60% of total suspended particles over the 12 months of the study, and their concentrations ranged from 19.3 to 41.7 microg/m(3). Almost all the PAHs were found in the PM2.5 fraction, in which the summed concentration (sigma PAH) of the 21 PAHs ranged from 1.0 to 8.4 ng/m(3). PAH concentrations were elevated in the colder seasons, but the concentrations of the PM2.5 fraction of suspended particles were not. The composition of PAHs associated with PM2.5 changed little during the year of our study, indicating that seasonal sources such as heating have little effect. Factor analysis showed that three factors explain little of the seasonal variations in the sources of PAHs. Contour maps showed high concentrations of indicator PAHs for gasoline and diesel engine sources distributed along major roads and near factories on the outskirts of the city. We concluded that the contribution of vehicle exhausts to particle-associated PAHs is significant not only in the central city, but also in outer areas associated with major ring roads, and suggest that air pollution in outskirts of urban area also is concerned for health risk associated with the exposure. PMID- 17129670 TI - Chlorination byproducts, their toxicodynamics and removal from drinking water. AB - No doubt that chlorination has been successfully used for the control of water borne infections diseases for more than a century. However identification of chlorination byproducts (CBPs) and incidences of potential health hazards created a major issue on the balancing of the toxicodynamics of the chemical species and risk from pathogenic microbes in the supply of drinking water. There have been epidemiological evidences of close relationship between its exposure and adverse outcomes particularly the cancers of vital organs in human beings. Halogenated trihalomethanes (THMs) and haloacetic acids (HAAs) are two major classes of disinfection byproducts (DBPs) commonly found in waters disinfected with chlorine. The total concentration of trihalomethanes and the formation of individual THM species in chlorinated water strongly depend on the composition of the raw water, on operational parameters and on the occurrence of residual chlorine in the distribution system. Attempts have been made to develop predictive models to establish the production and kinetics of THM formations. These models may be useful for operational purposes during water treatment and water quality management. It is also suggested to explore some biomarkers for determination of DBP production. Various methods have been suggested which include adsorption on activated carbons, coagulation with polymer, alum, lime or iron, sulfates, ion exchange and membrane process for the removal of DBPs. Thus in order to reduce the public health risk from these toxic compounds regulation must be inforced for the implementation of guideline values to lower the allowable concentrations or exposure. PMID- 17129671 TI - A summary of some experimental data on LNG safety. AB - In a recent paper, Luketa-Hanlin reviewed the information in the public domain related to LNG safety. The purpose of this paper is to supplement that work by providing a summary of the experimental information that Advantica has collected on LNG behaviour over the course of the last 30 years. This summary includes previously unpublished information obtained as a result of a number of collaborative projects. Subjective comments are also made on the status of modelling for each of the topic areas and, in a discussion, views are provided on those areas where there are currently gaps that may have a major impact on evaluating the individual or societal risks associated with LNG operations. PMID- 17129672 TI - Degradation of commercial azo dye reactive Black B in photo/ferrioxalate system. AB - The photolysis and photo-catalysis of ferrioxalate in the presence of hydrogen peroxide with UV irradiation (UV/ferrioxalate/H(2)O(2) process) for treating the commercial azo dye, reactive Black B (RBB), is examined. An effort is made to decolorize textile effluents at near neutral pH for suitable discharge of waste water. pH value, light source, type of initial catalyst (Fe(3+) or Fe(2+)) and concentration of oxalic acid (Ox) strongly affected the RBB removal efficiency. The degradation rate of RBB increased as pH or the wavelength of light declined. The optimal molar ratio of oxalic acid to Fe(III) is three, and complete color removal is achieved at pH 5 in 2h of the reaction. Applying oxalate in such a photo process increases both the RBB removal efficiency and the COD removal from 68% and 21% to 99.8% and 71%, respectively. PMID- 17129673 TI - Charge defects glowing in the dark. AB - We investigate the effects of local charge defects in HREM imaging, using electron densities calculated by density functional methods. As a model of a planar interface with a local charge defect we use the polar MgO square root (111)-3 x 3R30 degrees surface, which has an additional hole per surface unit cell. A complimentary example, the non-polar MgO (100) surface that has no local charge defect is simulated for comparison. We show that the contrast due to local charge defects is rather high, and suggest that they should be directly observable. PMID- 17129674 TI - Genetic resistance to Sarcocystis miescheriana in pigs following experimental infection. AB - Clinical and parasitological traits of Sarcocystis miescheriana differ in Pietrain and Meishan pigs. For further description and characterization of the genetic basis of this variation a F(2) family based on Pietrain boars and Meishan sows as founders was generated. One hundred and thirty-nine F(2) pigs were challenged orally at an age of 100 days with 50,000 sporozysts to produce the typical clinical picture of a moderate dose Sarcocystis infection. Heritabilities were estimated for clinical and clinical-chemical traits, for specific antibody responses to the infection and for bradyzoite numbers found in skeletal (Musculus longissimus dorsi: M.l.d.) and heart muscles at necropsy 70 days post-infection (p.i.) Apart from several low to moderate heritabilities, high heritabilities were observed for bradyzoite numbers in the M.l.d. (0.68), IgM antibody levels (0.74) during the acute (14 days p.i.) and titres of specific IgG antibodies (0.42) in the early stage of cyst formation (42 days p.i.). Marked heritabilities of these traits, which are basic for acute phase of the disease (14 days p.i.) or chronic Sarcocystosis presume genes that explain sufficient shares of variance (QTL). The model is considered valuable for screening of gene variants associated with resistance/susceptibility to Sarcocystis infection. Such gene variants could then be used in susceptibility-scoring or selection programs in the future. PMID- 17129675 TI - Evaluation of bithionol as a bath treatment for amoebic gill disease caused by Neoparamoeba spp. AB - This study examined the toxicity of bithionol to Atlantic salmon Salmo salar and rainbow trout Oncorhynchus mykiss in fresh- and seawater and the efficacy of bithionol as a 1h seawater bath treatment for amoebic gill disease (AGD). To examine toxicity, fish were bathed for 1, 3 and 6h in bithionol, an anti protozoal at 0, 1, 5, 10, 25 and 35mgL(-1) with toxicity determined by time to morbidity. Efficacy was examined by bathing AGD-affected Atlantic salmon and rainbow trout for 1h at bithionol concentrations of 1-25mgL(-1). Efficacy was determined by examining gill amoeba counts and identifying percent lesioned gill filaments at 1 and 24h after bath exposure to bithionol. For both species, bithionol was determined to be toxic at 25 and 35mgL(-1) exhibiting median lethal times (LT50s) ranging from 21 to 84min. Morbidity occurred in the 5 and 10mgL(-1) treatments, however, due to sampling regime there were not enough fish available to calculate LT50s. Only bithionol at 1mgL(-1) was considered non-toxic with no signs of morbidity. Bithionol was more toxic in seawater than freshwater and had no acute effects on gill Na+/K+ ATPase and succinic dehydrogenase, or plasma osmolality and chloride concentration. Bithionol at 1mgL(-1) reduced percent lesioned gill filaments in Atlantic salmon and rainbow trout by 33 and 27 per cent, respectively, compared to the seawater control. Similarly, numbers of amoeba were reduced by 33 and 43 per cent for Atlantic salmon and rainbow trout, respectively, when compared to the seawater control. Furthermore, bithionol reduced percent lesioned gill filaments as much as did the current industry standard of freshwater. This study demonstrated that a 1h seawater bath containing 1mgL(-1) bithionol could be an improvement to the current method of treatment for AGD-affected Atlantic salmon and rainbow trout. PMID- 17129676 TI - Detection and measurement of benzimidazole resistance alleles in Haemonchus contortus using real-time PCR with locked nucleic acid Taqman probes. AB - Benzimidazole resistance is a common problem in parasitic nematodes of ruminants and early detection is vital if its spread is to be monitored and controlled. Real time PCR offers a fast and reliable method for rapid detection and measurement of resistance allele frequencies. In Haemonchus contortus a single nucleotide polymorphism at codon 200 of the beta-tubulin gene (TTC to TAC), causing a phenylalanine to tyrosine amino acid substitution, has been shown to be involved in many cases of resistance. Locked nucleic acid (LNA) Taqman probes have been used in this work to detect and measure the frequency of resistance alleles in individual and multiple H. contortus. Detection of resistant genotypes using LNA Taqman probes in individual H. contortus is simpler and more reliable than with previously described assays. Measurement of the frequency of resistant alleles in populations of H. contortus was achieved by using the cycle threshold (C(t)) values and a standard curve derived from populations with known allele frequencies. Results using the LNA probes on individual and multiple worms gave similar results to the allele specific PCR. The sensitivity of the LNA assay on multiple nematodes allowed reliable detection of > or = 10% resistance allele frequency. Using the final fluorescence method, it was possible to differentiate populations with approximately 0, 5 and 10% resistance allele frequencies. PMID- 17129677 TI - Expression of the O-linked N-acetylglucosamine containing epitope H in normal myometrium and uterine smooth muscle cell tumors. AB - Epitope H contains an O-linked N-acetylglucosamine (O-GlcNAc) residue in a specific conformation and/or environment recognized by monoclonal antibody H (mAbH). We have previously shown that epitope H is present in more than one polypeptide and in various types of normal and pathological cells. In the present study, we focused on uterine smooth muscle cell tumors and their adjacent normal myometrium to gain further insight into the expression patterns of epitope H in human tissues. The indirect immunoperoxidase method was applied using the mAbH and the monoclonal anti-cytokeratin 8 antibody (AbCK8) in 50 cases of typical uterine leiomyomas and in five cases of uterine leiomyosarcomas, with four cases belonging to Group II A and one to Group III according to Bell et al. [6]. Western immunoblotting was applied using mAbH and AbCK8 in five cases of uterine leiomyomas and their adjacent myometrium. The main results were as follows: (1) epitope H showed intense immunohistochemical expression in 46% (23/50) and moderate expression in 54% (27/50) of uterine leiomyomas, (2) epitope H showed intense immunohistochemical expression in 40% (2/5) and moderate expression in 60% (3/5) of uterine leiomyosarcomas, (3) epitope H showed no difference in the immunohistochemical expression between leiomyomas and their adjacent myometrium and between leiomyosarcomas and their adjacent myometrium, (4) immunohistochemical expression of cytokeratin 8 was not detected in the normal and neoplastic smooth muscle cells, (5) Western immunoblotting showed that in the smooth muscle cells of the myometrium and leiomyomas, epitope H is localized in four polypeptides with molecular weights of 100, 61, 59, and 54 kDa, and (6) Western immunoblotting did not detect cytokeratin 8 in the normal and neoplastic smooth muscle cells. The present results indicate fluctuations of the epitope expression levels in uterine smooth muscle cell tumors and their adjacent myometrium. These fluctuations may be of interest for gaining insight into the pathogenesis of uterine smooth muscle cell tumors, since O-GlcNAc glycosylation is involved in cell cycle and apoptosis pathways and may modify proteins involved in oncogenesis (tumor suppressor proteins and oncoproteins) and proteins with important biological functions such as cytoskeletal proteins, transcription factors, and heat-shock proteins. Furthermore, the present results indicate that cytokeratin 8, without being present in the cells of the myometrium, leiomyomas and leiomyosarcomas, shares its epitope H, which contains its unique sugar O-N acetylglucosamine residue, with four other unrelated polypeptides produced by the normal and neoplastic smooth muscle cells. This should be considered when using anti-cytokeratin 8 antibodies in immunohistochemistry against smooth muscle cell tumors to avoid false positive immunohistochemical results. PMID- 17129678 TI - Coexistence between Cyphomyrmex ants and dominant populations of Wasmannia auropunctata. AB - The little fire ant Wasmannia auropunctata is able to develop highly dominant populations in disturbed areas of its native range, with a resulting negative impact on ant diversity. We report here on the tolerance of such populations towards several fungus-growing ants of the genus Cyphomyrmex (rimosus complex) in French Guiana. This tolerance is surprising given the usually high interspecific aggressiveness of W. auropunctata when dominant. In order to understand the mechanisms behind such proximity, aggressiveness tests were performed between workers of the different species. These behavioural assays revealed a great passivity in Cyphomyrmex workers during confrontations with W. auropunctata workers. We also found that the aggressiveness between W. auropunctata and two Cyphomyrmex species was more intense between distant nests than between adjacent ones. This dear-enemy phenomenon may result from a process of habituation contributing to the ants' ability to coexist over the long term. PMID- 17129679 TI - Interval bisection in spontaneously hypertensive rats. AB - An interval bisection procedure was used to study time discrimination in spontaneously hypertensive rats (SHR), which have been proposed as an animal model for the attention deficit hyperactivity disorder (ADHD); Wistar Kyoto and Wistar rats were used as comparison groups. In this procedure, after subjects learn to make one response (S) following a short duration stimulus, and another (L) following a long duration stimulus, stimuli of intermediate durations are presented, and the percentage of L is calculated for each duration. A logistic function is fitted to these data, and different parameters that describe the time discrimination process are obtained. Four conditions, with different short and long durations (1-4, 2-8, 3-12, 4-16s) were used. The results indicate that time discrimination is not altered in SHR, given that no difference in any of the parameters obtained were significant. Given that temporal processing has been proposed as a fundamental factor in the development of the main symptoms of ADHD, and that deficits in time discrimination have been found in individuals with that disorder, the present results suggest the necessity of exploring time perception in SHR with other procedures and sensory modalities, in order to assess its validity as an animal model of ADHD. PMID- 17129680 TI - The Network for the Improvement of Addiction Treatment (NIATx): enhancing access and retention. AB - The Network for the Improvement of Addiction Treatment (NIATx) teaches participating treatment centers to use process improvement strategies. A cross site evaluation monitored impacts on days between first contact and first treatment and percent of patients who started treatment and completed two, three and four units of care (i.e., one outpatient session, 1 day of intensive outpatient care, and 1 week of residential treatment). The analysis included 13 agencies that began participation in August 2003, submitted 10-15 months of data, and attempted improvements in outpatient (n=7), intensive outpatient (n=4) or residential treatment services (n=4) (two agencies provided data for two levels of care). Days to treatment declined 37% (from 19.6 to 12.4 days) across levels of care; the change was significant overall and for outpatient and intensive outpatient services. Significant overall improvement in retention in care was observed for the second unit of care (72-85%; 18% increase) and the third unit of care (62-73%; 17% increase); when level of care was assessed, a significant gain was found only for intensive outpatient services. Small incremental changes in treatment processes can lead to significant reductions in days to treatment and consistent gains in retention. PMID- 17129681 TI - Stress- and cue-induced cigarette craving: effects of a family history of smoking. AB - Individuals with multiple smokers among first-degree relatives (FH+) are significantly more likely to be persistent smokers themselves. The mechanisms underlying this relationship are unknown. An independent line of research has suggested that persistent smoking is more common among smokers with heightened levels of cigarette craving after being exposed to smoking cues and stressors. The present study experimentally tested the hypothesis that FH+ smokers would exhibit stronger stress- and cue-induced craving reactions compared to FH- smokers. We also explored gender and ethnicity-related differences in these effects. To that end, 160 smokers were recruited by advertisement and exposed to neutral (changing a light bulb), stressful (dental work), and smoking (lighting up after a meal) situations, using script-guided imagery under controlled laboratory conditions. Participants completed craving questionnaires before and after each condition. Supporting the hypotheses, even after controlling smoking history and strength of habit, FH+ smokers (n=86) displayed stronger craving reactions to both dental and smoking imagery (p's<0.05) than FH- smokers (n=74). Interestingly, women had higher stress-, but not smoking cue-induced cravings, than men, with FH+ women exhibiting the highest levels of stress-induced craving. Findings suggest a mechanism through which a family history of smoking leads to poorer cessation success, especially among women. PMID- 17129682 TI - Theory-based active ingredients of effective treatments for substance use disorders. AB - This paper describes four related theories that specify common social processes that protect individuals from developing substance use disorders and may underlie effective psychosocial treatments for these disorders: social control theory, behavioral economics and behavioral choice theory, social learning theory, and stress and coping theory. It then provides an overview of the rationale and evidence for four effective psychosocial treatments for substance use disorders: motivational interviewing and motivational enhancement therapy, 12-step facilitation treatment, cognitive-behavioral treatment and behavioral family counseling, and contingency management and community reinforcement approaches. The presumed active ingredients of these treatments are described in terms of how they exemplify the social processes highlighted by the four theories. The identified common components of effective treatment include support, goal direction, and structure; an emphasis on rewards that compete with substance use, a focus on abstinence-oriented norms and models, and attempts to develop self efficacy and coping skills. Several issues that need to be addressed to enhance our understanding of the active ingredients involved in effective treatment are discussed, including how to develop measures of these ingredients, how well the ingredients predict outcomes and influence conceptually comparable aspects of clients' life contexts, and how much their influence varies depending upon clients' demographic and personal characteristics. PMID- 17129683 TI - Psychiatric disorders in inhalant users: results from The National Epidemiologic Survey on Alcohol and Related Conditions. AB - OBJECTIVE: To examine the prevalence and correlates of mood, anxiety, and personality disorders among lifetime inhalant users. METHODS: Statistical analyses were based on data from the 2001-2002 National Epidemiologic Survey on Alcohol and Related Conditions (NESARC), a nationally representative survey of adults in the United States. RESULTS: Inhalant users (N=664) had high lifetime prevalences of DSM-IV mood (48%), anxiety (36%), and personality (45%) disorders. Of all inhalant users, 70% met criteria for at least one lifetime mood, anxiety, or personality disorder and 38% experienced a mood or anxiety disorder in the past year. Prevalences of comorbid psychiatric disorders varied by gender. Compared with male inhalant users, female inhalant users had higher prevalences of lifetime dysthymia (24% versus 16%), any anxiety disorder (53% versus 30%), panic disorder without agoraphobia (25% versus 11%), and specific phobia (28% versus 14%), but a lower prevalence of antisocial personality disorder (22% versus 36%). Female inhalant users also were more likely than male inhalant users to meet criteria for three or more mood or anxiety disorders (15% versus 8%) in the past year. Among inhalant users with comorbid disorders, those who developed social or specific phobia typically experienced onset of these disorders prior to initiation of inhalant use; all other mood and anxiety disorders usually developed following the onset of inhalant use. Inhalant users who were women, poor, less educated, with early onset of inhalant use, family histories of psychopathology, and personal histories of substance abuse treatment had increased odds of psychiatric disorders. CONCLUSIONS: Psychiatric disorders are highly prevalent among inhalant users nationally and female inhalant users are more likely than male inhalant users to experience multiple psychiatric disorders. Inhalant use and its consequences among females warrant greater research attention. PMID- 17129684 TI - Education inequality and use of cigarettes, alcohol, and marijuana. AB - Education inequality at the neighborhood-level may influence population health and health behavior. We assessed the relations between education inequality and substance use in 59 New York City (NYC) neighborhoods. We used Gini coefficients of education to describe neighborhood education inequality and data from a random digit-dial phone survey of adult residents of NYC to assess use of substances. Among 1355 respondents (female=56.2%; white=35.7%; mean age=40.4), 23.9% (95% confidence interval [CI]=20.3-27.5) reported smoking, 39.4% (95% CI=35.3-43.4) drinking, and 5.4% (95% CI=3.6-7.3) using marijuana in the previous 30 days. In multilevel models controlling for neighborhood education, neighborhood income inequality, and individual covariates, living in a neighborhood with high education inequality was associated with a greater prevalence of drinking (p=0.02) and of smoking marijuana (p=0.004) but among current drinkers it was associated (p=0.03) with having fewer drinks. The odds of alcohol use (OR=1.70) and marijuana use (OR=3.49) were greater in neighborhoods in the 75th percentile of education Gini compared to neighborhoods in the 25th percentile of education Gini. Statisical interactions suggest that there may be a stronger relation between education inequality and marijuana use in neighborhoods with low mean education than in neighborhoods with higher mean levels of education. These findings, taken together, suggest a complex relation between education inequality and substance use; likelihood of the use of alcohol and marijuana was higher in areas with higher education inequality suggesting potential roles for substance use norms and availability, whereas quantity used among drinkers was higher in areas with low education inequality, suggesting potential roles for both disadvantage and norms. PMID- 17129685 TI - A deep-branching clade of retrovirus-like retrotransposons in bdelloid rotifers. AB - Rotifers of class Bdelloidea, a group of aquatic invertebrates in which males and meiosis have never been documented, are also unusual in their lack of multicopy LINE-like and gypsy-like retrotransposons, groups inhabiting the genomes of nearly all other metazoans. Bdelloids do contain numerous DNA transposons, both intact and decayed, and domesticated Penelope-like retroelements Athena, concentrated at telomeric regions. Here we describe two LTR retrotransposons, each found at low copy number in a different bdelloid species, which define a clade different from previously known clades of LTR retrotransposons. Like bdelloid DNA transposons and Athena, these elements are found preferentially in telomeric regions. Unlike bdelloid DNA transposons, many of which are decayed, the newly described elements, named Vesta and Juno, inhabiting the genomes of Philodina roseola and Adineta vaga, respectively, appear to be intact and represent recent insertions, possibly from an exogenous source. We describe the retrovirus-like structure of the new elements, containing gag, pol, and env-like open reading frames, and discuss their possible origins, transmission, and behavior in bdelloid genomes. PMID- 17129686 TI - Evidence for trans-border movement of rabies by wildlife reservoirs between countries in the Balkan Peninsular. AB - Rabies remains endemic in a number of reservoir species throughout southeast Europe. To investigate the relationship between rabies viruses within this region we have compared rabies virus nucleoprotein sequence data from a diverse panel of rabies samples from Bulgaria with available data from countries in the Balkan Peninsula. This analysis provides a first description of rabies isolates in Bulgaria and suggests that there is evidence for wildlife-mediated movement of rabies with countries to the west of Bulgaria including Serbia and Bosnia Herzegovina but no evidence for a link with rabies foci in Turkey. We speculate that natural barriers such as the Balkan mountains may play a significant role in constraining the southerly spread of rabies in southeast Europe. PMID- 17129687 TI - Abundance and phenotypic diversity of Escherichia coli isolates with diminished susceptibility to expanded-spectrum cephalosporins in faeces from healthy food animals after slaughter. AB - Antimicrobial resistance (AR) is an increasing phenomenon but its quantitative estimation remains controversial. The classical resistance percentage approach is not well suited to detect either emergence or low levels resistance. One option is to shift the focus from strains to hosts. This approach is applied to test for phenotypic diversity associated with diminished susceptibility to expanded spectrum cephalosporins (DSESC) in faecal Escherichia coli from healthy food animals in Spain. We performed E. coli enumeration in faecal samples of broilers (82 pooled samples) and pigs (80 pooled samples) at the slaughterhouse level, using Coli-ID plates alone and supplemented with cefotaxime at two levels (1 and 8 microg/ml). Antimicrobial susceptibility of isolates was tested by the agar diffusion method. Clustering was carried out using these numerical values and Ward and UPGMA methods. When using plates supplemented with 1 microg/ml of cefotaxime for DSESC E. coli detection, 93% (76/82) of broiler pooled samples and 36% (29/80) pig pooled samples tested positive. When using 8 microg/ml of cefotaxime, 67% (55/82) of broilers and 13% (10/80) of pigs were positive. Nevertheless, the relative abundance of this phenotype was low in both animal species (range 0-4.3%). Irrespective of the clustering method (Ward or UPGMA), a noticeable phenotypic diversity was detected, especially from the plates containing 1 microg/ml of cefotaxime. We concluded that: (a) E. coli with phenotype DSESC are common in broilers and pigs but are less frequent in pigs, and (b) the host approach is the most appropriate method for antimicrobial resistance assessment when null or very low levels of antimicrobial resistant bacteria are expected. PMID- 17129688 TI - Are abnormal fidgety movements an early marker for complex minor neurological dysfunction at puberty? AB - BACKGROUND: Prechtl's method on the qualitative assessment of general movements (GMs) is a powerful tool for early and specific prediction of cerebral palsy. However, it is uncertain whether the GM assessment can be used to predict mild neurological impairment. AIMS: To determine whether the quality of general movements (GMs) from the age of 3 to 5 months, i.e. fidgety movements, is related to the presence of complex minor neurological dysfunctions (MND) 13 to 15 years later. STUDY DESIGN: Prospectively collected data on the quality of GMs during infancy were retrospectively analysed on the basis of MND at puberty. SUBJECTS: Twenty-eight participants (14 girls and 14 boys) with a median gestational age of 40 weeks (range: 35 to 42 weeks) and an appropriate birth weight (median 3390 g; range 1900 to 4200 g). OUTCOME MEASURES: Touwen's neurological examination. RESULTS AND CONCLUSIONS: Abnormal fidgety movements were not related to later complex MND, but to fine manipulative disabilities (p<0.05). Normal fidgety movements, which are continually present in the whole body, might be required for optimal calibration of the proprioceptive system. PMID- 17129689 TI - Diallyl sulfide induces the expression of estrogen metabolizing genes in the presence and/or absence of diethylstilbestrol in the breast of female ACI rats. AB - Diethylstilbestrol (DES) induces mammary tumors in female ACI rats and is associated with an increased risk of developing breast cancer in humans. Diallyl sulfide (DAS) has been shown to prevent cancer in animals. Previously, we have shown that DAS inhibits the production of DES induced DNA adducts when given prior to DES. We hypothesize that DAS alters the expression of genes responsible for DES metabolism. To test this hypothesis, four groups of 10 female ACI rats were treated daily for four days as follows: (1) corn oil, (2) 50mg/kg DES, (3) 50mg/kg DAS, and (4) 50mg/kg DAS+50mg/kg DES. RNA was isolated from breast tissue and mRNA levels of CYP1A1, CYP1B1, glutathione-S-transferase (GST) and superoxide dismutase (SOD) were analyzed by real-time PCR. DES, DAS, and DES/DAS treatments increased the expression of CYP1A1 by 2.1-, 4.7-, and 12.7-fold, respectively. Similar results were seen for CYP1B1. DES decreased the expression of GST by 23%, whereas DAS and DAS/DES treatments increased the expression of GST by 12- and 16.7-fold, respectively. Similar results were seen with SOD. These results suggests that DAS may prevent the formation of DES induced DNA damage by altering the expression of DES metabolizing genes. PMID- 17129690 TI - The effect of oxidative stress on macrophages and lung epithelial cells: the role of phosphodiesterases 1 and 4. AB - Reactive oxygen species (ROS) have been implicated in various pulmonary diseases by causing direct injury to lung epithelial cells. Signalling activity of cells through transcription factors such as nuclear factor kappa B (NF-kappaB) and AP-1 have been shown to be regulated by ROS, and the release of pro-inflammatory cytokines demonstrated in the study of inflammatory disease. In this study, we examined the effect of the oxidant tert-butylhydroperoxide (tBHP) on mouse J774 macrophages and its ability to cause the release of the pro-inflammatory cytokine tumour necrosis factor alpha (TNF-alpha). The role of calcium as a signalling molecule was studied using various calcium antagonists. The role of the signalling molecule cAMP was also investigated using phosphodiesterase inhibitors PDE1 and PDE4 families. Oxidative stress was investigated in lung epithelial (A549) cells with and without calcium antagonists and PDE inhibitors with regard to their ability to modulate release of the neutrophil chemoattractant interleukin 8 (IL-8). The oxidant tBHP significantly increased the cytosolic calcium concentration in J774 macrophages, which was prevented by the PDE1 inhibitor. The production of TNF-alpha protein by J774 macrophages was mediated by a pathway involving calcium as addition of calcium antagonists inhibited the tBHP stimulated increase in the cytokine. Inhibitors of both PDE1 and PDE4 completely prevented the tBHP stimulated TNF-alpha release suggesting that the cAMP pathway may be important in the oxidant induced signalling pathway leading to gene expression of pro-inflammatory cytokines. In the presence of oxidant alone, A549 epithelial cells released significant amounts of IL-8, which was inhibited by both calcium antagonist treatment and PDE inhibition treatment. These data suggest that ROS-mediated lung inflammation could be mediated at least in part by calcium and elevated PDE activity associated with decreased cAMP in both macrophages and epithelial cells. Inhibition of these pathways may provide a route for treatment of inflammatory lung diseases. PMID- 17129691 TI - Drug hydrate systems and dehydration processes studied by terahertz pulsed spectroscopy. AB - Terahertz pulsed spectroscopy was used to distinguish between different hydrate systems. In the example of four pharmaceutical materials lactose, carbamazepine, piroxicam and theophylline it was demonstrated that all different hydrate and anhydrate forms exhibit distinct spectra in the far infrared. Furthermore the dehydration of theophylline monohydrate was characterised in situ. Here, a phase transition from the monohydrate to the anhydrous form was observed, followed by evaporation of the hydrate water in a second step. The rotational spectrum of water vapour is very characteristic in the far infrared and can easily be discerned from the terahertz spectrum of the solid state form. PMID- 17129692 TI - The effect of oil components on the physicochemical properties and drug delivery of emulsions: tocol emulsion versus lipid emulsion. AB - An emulsion system composed of vitamin E, coconut oil, soybean phosphatidylcholine, non-ionic surfactants, and polyethylene glycol (PEG) derivatives (referred to as the tocol emulsion) was characterized in terms of its physicochemical properties, drug release, in vivo efficacy, toxicity, and stability. Systems without vitamin E (referred to as the lipid emulsion) and without any oils (referred to as the aqueous micelle system) were prepared for comparison. A lipophilic antioxidant, resveratrol, was used as the model drug for emulsion loading. The incorporation of Brij 35 and PEG derivatives reduced the vesicle diameter to <100nm. The inclusion of resveratrol into the emulsions and aqueous micelles retarded the drug release. The in vitro release rate showed a decrease in the order of aqueous micelle system>tocol emulsion>lipid emulsion. Treatment of resveratrol dramatically reduced the intimal hyperplasia of the injured vascular wall in rats. There was no significant difference in this reduction when resveratrol was delivered by either emulsion or the aqueous micelle system. The percentages of erythrocyte hemolysis by the emulsions and aqueous micelle system were approximately 0 and approximately 10%, respectively. Vitamin E prevented the aggregation of emulsion vesicles. The mean vesicle size of the tocol emulsion remained unchanged during 30 days at 37 degrees C. The lipid emulsion and aqueous micelle system, respectively, showed 11- and 16-fold increases in vesicle size after 30 days of storage. PMID- 17129693 TI - Anti-inflammatory and anti-angiogenic activities of Gastrodia elata Blume. AB - Gastrodia elata Blume rhizome has been traditionally used as a folk medicine for centuries in Oriental countries. Its ethanol extract (GEE) and subsequent fractions were used to evaluate anti-angiogenic, anti-inflammatory and related activities of Gastrodia elata. GEE potently inhibited angiogenesis in the chick chorioallantoic membrane assay, and its n-butanol fraction (BuOH) exerted the higher inhibitory effect. In a dose-dependent manner, GEE inhibited vascular permeability induced by acetic acid. GEE and its BuOH fraction exerted an inhibitory activity on exudate production, leukocyte migration and nitric oxide (NO) level in rat air-pouch model. GEE caused a dose-dependent inhibition of acetic acid-induced abdominal writhing in mice. In addition, GEE inhibited NO production and expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) upon stimulation by lipopolysaccharide (LPS) in RAW264.7 macrophages. In summary, we demonstrate some novel pharmacological activities of Gastrodia elata, such as anti-angiogenic, anti-inflammatory and analgesic activities, and in vivo and in vitro inhibitory activity on NO production. PMID- 17129694 TI - Effect of ethanol on cardiac single sodium channel gating. AB - Alcohol in modest and higher doses has the potential to induce cardiac arrhythmias. The most famous alcohol-related arrhythmia is the "holiday heart syndrome". Furthermore, there is a clear association between excessive alcohol consumption and the risk of sudden cardiac death. However, the acute effects of ethanol on arrhythmia induction are not well understood. The effect of ethanol on single cardiac sodium channels has not been studied yet. To elucidate the effect of ethanol on human cardiac sodium channels we performed a patch clamp study in HEK-293 cells overexpressing the human cardiac sodium channel. We used HEK-293 cells overexpressing the human cardiac sodium channel (Na(1.5)). Single channel gating was investigated by the cell-attached patch clamp technique. Sodium channel currents were elicited by depolarizing pulses from -120 to -20mV for a duration of 150ms. Single channel availability, open probability and peak average current were assessed baseline and after addition of ethanol in increasing concentrations (0.50 per thousand (10.9mM), 1.00 per thousand (21.7mM), 2.00 per thousand (43.5mM) and 4.00 per thousand (87.0mM)). We found a concentration dependent reduction of open probability which was statistically significant at 2.00 per thousand ethanol (66.5+/-14% of control). At higher concentrations (4.00 per thousand) also availability decreased to 66.5+/-11.0% of control. This resulted in a significant decrease of peak average current at 2.00 per thousand and at 4.00 per thousand ethanol (61.8+/-7.4 and 53.0+/-8.2% of control). For the first time the present study demonstrates acute inhibitory effects of ethanol on single cardiac sodium channel gating and provides one potential mechanism for the well known clinical observation that ethanol triggers supraventricular and ventricular arrhythmias. PMID- 17129695 TI - Tonsillar cyst of the false vocal cord. AB - OBJECTIVES/HYPOTHESIS: To report, for the first time, tonsillar cyst of the false vocal cord. STUDY DESIGN: Case report. METHODS: Case presentation and literature review. RESULTS: A 74-year-old woman with a mass in the false vocal cord presented with hoarseness. Histopathological examination revealed tonsillar cyst. Marsupialization via microlaryngeal approach failed. Finally the cyst was resected successfully through a lateral cervical approach with concomitant tracheotomy. CONCLUSIONS: This is the first report of tonsillar cyst in the false vocal cord in English literature. The cyst was finally resected through the lateral cervical approach with success. PMID- 17129696 TI - Oncogenic role of Epstein-Barr virus-encoded small RNAs (EBERs) in nasopharyngeal carcinoma. AB - OBJECTIVE: Epstein-Barr virus (EBV) is a causative agent of nasopharyngeal carcinoma (NPC), and EBV gene expression is considered to be closely associated with the pathogenesis of NPC. Among EBV genes expressed in NPC, EBV-encoded non polyadenylated RNAs, termed EBERs, are the most abundant transcripts of EBV in NPC. However, the role of EBERs still remains unclear. This study was designed to investigate the relevance of EBERs to the oncogenesis of NPC. METHODS: Two types of EBERs expression vectors (EBERs-high-expression vector and EBERs-low expression vector) were constructed and transfected into EBV-negative cells, MDCK or EBV-negative clones of NPC-KT cells. Then, malignant transformation, represented by anchor independent growth, was evaluated between the EBERs transfected cells and EBERs-negative cells using a soft agar colony formation assay. Apoptosis was induced by serum deprivation (0.1% concentration of fetal bovine serum) and interferon-alpha (IFN-alpha) (500 U/ml) treatment. Cell viability was evaluated with a trypan blue exclusion test. The activation of cellular transcriptional factor NF-kappaB was studied with the IL-8 promoter sequence using a luciferase reporter assay. RESULTS: EBERs-high-expression vector transfected MDCK cells showed enhanced growth ability in soft agar compared with either EBERs-low-expression vector-transfected MDCK cells or EBERs-untransfected MDCK cells. However, they did not show the acquisition of any anti-apoptotic potential against either IFN-alpha or serum deprivation. Introduction of EBERs low-expression vector into MDCK cells did not show anchor independent growth characteristics. Neither EBV-negative NPC-KT cells nor MDCK cells transfected with EBERs-high-expression vector showed any difference from EBERs-untransfected EBV-negative NPC-KT cells. Introduction of EBERs into MDCK cells did not transactivate the IL-8 promoter, indicating that neither NF-kappaB nor AP-1 was activated by EBERs. CONCLUSION: EBERs are believed to induce the initial transformation of epithelial cells, thus contributing to the oncogenesis of NPC. Expression of abundant EBERs is considered to be critical for this transforming property of EBERs. PMID- 17129697 TI - Quantitative and qualitative determination of six xanthones in Garcinia mangostana L. by LC-PDA and LC-ESI-MS. AB - A new method was developed for the simultaneous analysis of six naturally occurring xanthones (3-isomangostin, 8-desoxygartanin, gartanin, alpha-mangostin, 9-hydroxycalabaxanthone and beta-mangostin). The quantitative determination was conducted by reversed phase high performance liquid chromatography with photodiode array detector (LC-PDA). Separation was performed on a Phenomenex Luna C18(2) (150 mm x 3.00 mm, 5 microm) column. The xanthones were identified by retention time, ultraviolet (UV) spectra and quantified by LC-PDA at 320 nm. The precision of the method was confirmed by the relative standard deviation (R.S.D.), which was < or =4.6%. The recovery was in the range from 96.58% to 113.45%. A good linear relationship was established in over two orders of magnitude range. The limits of detection (LOD) for six xanthone compounds were < or =0.248 microg/mL. The identity of the peaks was further confirmed by high performance liquid chromatography with time-of-flight mass spectrometry (LC-TOF MS) system coupled with electrospray ionization (ESI) interface. The developed methods were applied to the determination of six xanthones in Garcinia mangostana products. The satisfactory results showed that the methods are effective for the analysis of real samples. PMID- 17129698 TI - A rapid and simple HPLC method for the analysis of propofol in biological fluids. AB - A selective and sensitive high-performance liquid chromatographic method for the analysis of propofol in biological samples was developed. Propofol and thymol (internal standard) were analysed on a Purospher RP-18 endcapped (75 mmx4 mm, 3 microm) stationary phase using acetonitrile and water (65:35, v/v) as eluents at a flow rate of 0.6 mL/min. The excitation and emission wavelengths were 276 and 310 nm, respectively. Sample treatment consisted of deproteinization by acetonitrile containing the internal standard and direct injection of the supernatant. Mean analytical recovery were 105% (CV 2.0%) at concentrations ranging from 0.05 to 10 mg/L. The quantification limit was 3 ng/mL for a 500 microL sample plasma volume and 5 ng/mL for a 500 microL blood sample. The intra day and inter-day precisions were lower than 5.5% for three concentrations assessed (0.05, 1.0 and 10.0 mg/L). Considering the column size and the flow rate, the separation was achieved with an analysis time less than 6 min with a reduced consumption of solvent. This rapid HPLC method using a simple treatment procedure is sensitive enough for monitoring propofol in human biological samples. PMID- 17129699 TI - Lipid metabolism and cellular features of skeletal muscle and subcutaneous adipose tissue in pigs differing in IGF-II genotype. AB - In pigs, a paternally (pat) imprinted mutation in the IGF-II gene is associated with increased muscle mass and decreased backfat thickness. The aim of this study was to determine whether this mutation influenced cellular, biochemical and metabolic features of skeletal muscle and adipose tissue. Muscle (trapezius) and subcutaneous adipose tissue (SCAT) were collected from pigs (106kg) carrying (Qpat, n=6) or not carrying (qpat, n=7) the mutation. Adipocytes were isolated from those tissues by collagenase treatment. Lipid content and activity of lipogenic enzymes were determined using standard assays. Gene expression levels were determined by real-time PCR. Levels of IGF-II mRNA were higher (P<0.01) in muscle of Qpat than in that of qpat pigs, but they did not differ significantly between the two groups in SCAT. Whereas levels of IGF-I mRNA in muscle were similar in both groups, they were higher (P<0.05) in SCAT of Qpat pigs than in that of qpat pigs. Muscle lipid content and intramuscular adipocyte diameters were not influenced significantly by the IGF-II genotype. In SCAT, the reduction of backfat thickness in Qpat pigs compared with qpat pigs was associated with lower (P<0.05) lipid content and smaller (P<0.05) adipocytes, with no significant genotype-effects on expressions and/or activities of lipogenic enzymes. In summary, our results suggest that the IGF-II mutation altered body composition in pigs by favoring myofiber hypertrophy and repressing adipose cell development in SCAT. PMID- 17129700 TI - Primary malignant retroperitoneal tumors: analysis of a single institutional experience. AB - AIMS: In order to achieve complete resection in the surgical management of retroperitoneal tumors, it is crucial to know the tumor's anatomical location relative to neighboring organs. METHODS: Forty-nine patients with primary malignant retroperitoneal tumors were divided by tumor location into two groups [upper abdomen (group 1) or lower abdomen (group 2)], and clinicopathological features, tumor recurrence, and patient survival were assessed. RESULTS: No significant differences in preoperative clinical characteristics existed between two groups, and liposarcoma was the most frequently observed tumor type. The difference in the rates of complete resection between the two groups was not statistically significant (75.9% for group 1 and 85% for group 2). En-bloc combined resection was performed in 52% and 30% of patients in groups 1 and 2, respectively. The local recurrence rate in group 2 (31.3%) was higher than that in group 1 (9.5%), despite the fact that the differences in rates of complete resection and distant recurrence rates (14.3% in group 1 and 12.5% in group 2) between the two groups were not statistically significant. The overall 5-year survival rates were 67.9% for group 1 and 43.2% for group 2 (p=0.038). The 5-year survival rate of patients with tumors smaller than 10 cm was 78.4%, while that of patients with tumors larger than 10 cm was 38.1% (p=0.017). The 5-year survival rate after complete excision was 61%, whereas that after incomplete resection or biopsy only was 40.0% (p<0.0001). CONCLUSIONS: An upper abdominal tumor location is a positive prognostic factor even if small tumor size (<10 cm) and complete resection of the tumor are still more important factors to improve outcome in patients with malignant primary retroperitoneal tumors. Because complete resection was shown to be the most important prognostic factor, an aggressive and careful surgical approach is recommended for the treatment of such tumors. PMID- 17129701 TI - Comparison of clinical staging systems in predicting survival of hepatocellular carcinoma patients receiving major or minor hepatectomy. AB - AIM: To compare the utility of seven commonly used staging systems in the prediction of survival among patients with hepatocellular carcinoma (HCC) undergoing major or minor hepatectomy. METHODS: All patients were classified by the Okuda, the TNM, the CLIP, the BCLC, the CUPI, the JIS and the MELD classifications to estimate the probabilities of survival. Survival curves were calculated using the Kaplan-Meier method and were examined using log-rank testing. The overall predictive power for patient survival with each staging system was evaluated using linear trend chi(2) tests and from the area under the receiver operating characteristic (ROC) curve. RESULTS: In our patient cohort, the log-rank test and the linear trend chi(2) test of the CLIP and JIS systems gave better results than did the other staging systems. The discriminatory ability of the CLIP and JIS staging for death, evaluated by ROC curve areas, was also better. In the subgroups of major hepatectomy patients with a non-cirrhotic liver or minor hepatectomy patients with a cirrhotic liver, the CLIP and JIS systems showed similar better performances in these three tests. The discriminatory ability of the CLIP system was the best in major hepatectomy patients with a non-cirrhotic liver while JIS score discriminated best in minor hepatectomy patients with a cirrhotic liver. CONCLUSION: Among the seven staging systems, the CLIP and JIS systems perform better than do the others. While the CLIP system should be considered to stage major hepatectomy patients, the JIS system could be chosen to stage minor hepatectomy patients. PMID- 17129702 TI - Differential expression of manganese superoxide dismutase, copper/zinc superoxide dismutase, and catalase in gastric adenocarcinoma and normal gastric mucosa. AB - AIMS: The biologic significance of antioxidant enzymes (AOEs) in gastric adenocarcinoma is still unclear. The aims of this study was to investigate the differential expression of AOEs in gastric carcinoma cells and non-cancerous counterparts and the relationship with the various clinicopathologic variables in gastric cancer patients. METHODS: Expression status of MnSOD, Cu/ZnSOD, and catalase was evaluated immunohistochemically in 120 paired gastric cancer and adjacent non-cancerous tissue. The tissues were fixed in absolute methanol immediately after surgical resection and immunohistochemistry was performed by microprobe system using tissuemicroarray slides. RESULTS: All AOEs revealed moderate to strong immunoreactivity in the parietal and intestinal metaplastic cells. Stromal cells in both cancer and non-cancerous tissue expressed MnSOD and catalase but Cu/ZnSOD. Immunoreactivity of MnSOD and catalase was increased in gastric carcinoma cells compared to their non-cancerous counterparts and revealed an association with intestinal type adenocarcinomas whereas immunoreactivity of Cu/ZnSOD did not reveal significant difference between cancer and non-cancerous mucosal cells. CONCLUSIONS: Expression of MnSOD, Cu/ZnSOD, catalas in gastric cancer cells and non-cancerous counterparts was different and increased MnSOD and possibly catalase may in part be responsible for the increased risk of intestinal type adenocarcinoma of the stomach. PMID- 17129703 TI - Sentinel lymph node biopsy for high risk cutaneous squamous cell carcinoma: case series and review of the literature. AB - AIMS: Cutaneous squamous cell carcinoma (SCC) is the second most common skin cancer. The metastatic potential is generally low. However, there are subgroups of patients at higher risk, for whom sentinel lymph node biopsy (SLNB) might be useful. SLNB might allow the timely inclusion of high risk patients in more aggressive treatment protocols, sparing at the same time node-negative patients the morbidity of potentially unnecessary therapy. Our aim was to introduce the concept of SLNB for patients with high risk cutaneous SCC. PATIENTS AND METHODS: We examined a consecutive series of high risk cutaneous SCC patients undergoing SLNB at our large dermatological hospital, and performed a literature review and pooled analysis of all published cases of SLNB for cutaneous SCC. RESULTS: Among the 22 clinically node-negative patients undergoing SLNB at our hospital, one patient (4.5%) showed a histologically positive sentinel node and developed recurrences during follow-up. Sentinel node-negative patients showed no metastases at a median follow-up of 17 months (range: 6-64). The incidence of positive sentinel nodes in previous reports ranged between 12.5% and 44.4%. Pooling together patients from the present and previous studies (total 83 patients), we calculated an Odds Ratio of 2.76 (95% CI 1.2-6.5; p=0.02) of finding positive sentinel nodes for an increase in tumor size from <2 cm to 2.1-3 cm to >3 cm. CONCLUSIONS: Our case series and the pooled analysis support the concept that SLNB can be performed for high risk cutaneous SCC. Prospective multicenter studies are needed to examine the role, utility and cost effectiveness of SLNB for this population. PMID- 17129704 TI - Clinicopathological and prognostic significance of vascular endothelial growth factors (VEGF)-C and -D and VEGF receptor 3 in invasive breast carcinoma. AB - AIMS: Vascular endothelial growth factors C and D (VEGF-C and VEGF-D) play a major role in lymphangiogenesis and activate VEGF receptor 3 (VEGFR-3). Our purpose was to study the clinicopathologic and clinical value of VEGF-C, VEGF-D and VEGFR-3 in invasive breast carcinoma. MATERIAL AND METHODS: Immunohistochemistry was performed in paraffin-embedded tissue specimens from 177 invasive breast carcinomas to detect the proteins VEGF-C, VEGF-D, VEGFR-3, p53, Ki67, c-erbB-2, topoII alpha and ER/PR. The results were statistically processed. RESULTS: VEGF-C, VEGF-D and VEGFR-3 were found to be predominantly expressed in the cytoplasm of the malignant cells. VEGF-C occasionally showed a submembranous intensification. VEGF-D and VEGFR-3 were also immunodetected in the nuclei of the malignant cells. Nuclear VEGF-D was positively correlated to p53, Ki67 and topoII alpha proteins' expression (p=0.003, p=0.009 and p=0.017 respectively) and nuclear VEGFR-3 to topoII alpha (p=0.034). Cytoplasmic expression of VEGF-C and its submembranous intensification were found to be independent indicators of patients' overall and disease-free survival, respectively (p=0.003 and p=0.044 respectively). The group with high expression of both cytoplasmic VEGF-C and stromal VEGFR-3 showed poor overall survival (p=0.024) and the group with both submembranous VEGF-C and stromal VEGFR-3 immunostaining showed poor both disease free and overall survival (p=0.012 and p=0.038 respectively). CONCLUSION: VEGF-D and VEGFR-3 seem to exert proliferative activity in invasive breast carcinomas. VEGF-C was found to be an independent indicator of patient's poor prognosis and the simultaneous expression of tumor VEGF-C and stromal VEGFR-3 yielded additional prognostic information. PMID- 17129705 TI - Changes in immune and glial markers in the CSF of patients with Complex Regional Pain Syndrome. AB - Complex Regional Pain Syndrome is a severe chronic pain condition characterized by sensory, autonomic, motor and dystrophic signs and symptoms. The pain in CRPS is continuous, it worsens over time, and it is usually disproportionate to the severity and duration of the inciting event. This study compares cerebrospinal fluid (CSF) levels of pro- and anti-inflammatory cytokines, chemokines and several biochemical factors (glial fibrillary acidic protein (GFAP), the nitric oxide metabolites (nitrate plus nitrite), the excitatory amino acid neurotransmitter glutamate, calcium, total protein and glucose) in patients afflicted with CRPS to levels found in patients suffering with other non-painful or painful conditions. The aim of the study is to determine the degree of involvement of glial cells and immune system mediators in the pathophysiology of CRPS. There was no elevation or reduction of a CSF marker that was specific for CRPS patients. However, there were several patterns of markers that could be helpful in both elucidating the mechanisms involved in the disease process and supporting the diagnosis of CRPS. The most common pattern was found in 50% (11 out of 22) of the CRPS patients and consisted of; elevated IL-6, low levels of IL 4 or IL-10, increased GFAP or MCP1 and increases in at least two of the following markers NO metabolites, calcium or glutamate. The results from this and other similar studies may aid in elucidating the mechanisms involved in the pathophysiology of CRPS. A better understanding of these mechanisms may lead to novel treatments for this very severe, life-altering illness. PMID- 17129706 TI - Adolescent reward system perseveration due to nicotine: studies with methylphenidate. AB - Previous data suggests adolescent nicotine exposure permanently disrupts reward systems. Behavioral pharmacological methods were used to assess the effects of adolescent nicotine exposure on methylphenidate (MPD) sensitivity and reward from adolescence to adulthood. For experiment one, testing was performed on adult mice exposed to nicotine (0.3 and 3.0 mg/kg, SC, M-F, b.i.d.) or saline during adolescence (PND 25-57). After a 28-day drug-free, time-off period, the locomotor effects (30 min, 30 cm traveled) of MPD (5, 10, 20, and 40 mg/kg, IP) were measured. Thereafter, mice underwent condition-place-preference testing (CPP). MPD (20 mg/kg) was paired with the subject's non-preferred side and saline with the preferred side. Conditioning sessions were conducted for 8 days with a drug free post-test on the day following the final conditioning session. A second experiment was conducted to determine if adolescent mice respond differently to MPD compared to adult mice. The study compared adolescent mice (PND 25-46) to adult mice (PND 77-98) using identical MPD testing procedures as in experiment one. Adult subjects exposed to nicotine during adolescence behaved remarkably similar to adolescent subjects receiving only MPD. Both nicotine exposed subjects and naive adolescents exhibited increased response to MPD's motor activating effects and a decreased response to MPD's rewarding effects. Taken together it appears that adolescent nicotine exposure retards the development of reward systems, thus, maintaining an adolescent state indefinitely, which could result in increased vulnerability to substance abuse problems throughout adulthood. PMID- 17129707 TI - Levels of evidence and journal impact factor in oral and maxillofacial surgery. AB - The aim of this study was to identify the relationship between quality of research, in terms of levels of evidence, and journal impact factor (IF), and to describe the pattern of publications in oral and maxillofacial surgery. All four major journals in this subject area only, and with a published IF, were included in the study. Latest published IF dated 2004 was chosen, and all articles related to its calculation were accessed and classified into four levels of evidence. Correlation between levels of evidence and IF was investigated and the pattern of publications was described. All eligible 932 published articles were analysed. None (0%) were level I evidence, 20 (2%) were Level II, 70 (8%) level III and 337 (40%) level IV; 465 (50%) articles were classified as non-evidence. IF ranged from 0.689 to 1.154. There were statistically significant correlations between levels of evidence and IF (rho=1.0, P<0.01). Among the 465 non-evidence articles, there were 219 (47%) case reports, 91 (20%) animal studies, 52 (11%) laboratory studies, 35 (8%) technical notes, 24 (5%) tutorial articles, and 16 (3%) reviews articles. PMID- 17129708 TI - Benign familial neonatal convulsions: always benign? AB - BACKGROUND: Benign familial neonatal convulsions (BFNC) is a rare autosomal dominant seizure disorder usually described to be characterized by a benign course, spontaneous remission and normal psychomotor development. The latter statement had come under consideration when a few case reports of families with less than favorable outcomes were published. METHODS: Since 1998 a total of 112 families suspected to have BFNC have been referred to our lab for genetic testing. Within this sample we identified private KCNQ2 mutations in 17 BFNC families. For 10 of those 17 families follow up information about the psychomotor development and the outcome were available. RESULTS: In 4 (40%) of the 10 families at least 1 affected individual showed delayed psychomotor development or mental retardation. Three of the four mutations were familial, while the fourth mutation was de novo. Mutations associated with an unfavorable outcome tended to be located within the functionally critical S5/S6 regions of the KCNQ2 gene. CONCLUSIONS: Our data raise the question if BFNC can indeed be described as a benign disorder, and which are the genetic and/or environmental factors that influence the outcome. PMID- 17129709 TI - Challenging DSM-IV criteria for hypomania: diagnosing based on number of no priority symptoms. AB - BACKGROUND: DSM-IV definition of hypomania of bipolar-II disorder (BP-II), which includes elevated/irritable mood change as core feature (i.e., it must always be present), is not based on sound evidence. STUDY AIM: Following classic descriptions of hypomania, was to test if hypomania could be diagnosed on the basis of its number (9) of DSM-IV symptoms, setting no-priority symptom. METHODS: Consecutive 422 depression-remitted outpatients were re-interviewed by a mood specialist psychiatrist using the Structured Clinical Interview for DSM-IV Axis I Disorders-Clinician Version [a semi-structured interview modified by Benazzi and Akiskal (J Affect Disord, 2003; J Clin Psychiatry, 2005) to improve the probing for BP-II] in a private practice. History of episodes of subthreshold (i.e., 2 or more symptoms) and threshold (i.e., meeting DSM-IV criteria of elevated mood plus at least 3 symptoms, or irritable mood plus at least 4) hypomania, lasting at least 2 days, and which were the most common symptoms during the episodes, were systematically assessed. RESULTS: Bipolar-II disorder (BP-II) patients (according to DSM-IV criteria, apart from hypomania duration) were 260, and major depressive disorder (MDD) patients were 162. Mood change was present in all BP-II by definition. The most common symptoms were overactivity, which was present in almost all BP-II, followed by elevated mood and racing thoughts. ROC analysis of the number of hypomanic symptoms predicting BP-II found that a cut point of 5 or more symptoms over 9 had the best combination of sensitivity (90%) and specificity (84%), and the highest figure of correctly classified (87%) BP-II. History of episodes of 5 or more hypomanic symptoms was met by almost all BP-II. LIMITATIONS: Single interviewer. CONCLUSIONS: Following classic descriptions of hypomania, not setting any priority among the three basic domains of hypomania (mood, thinking, behavior), results suggest that a cutoff number of 5 symptoms over 9 (of those listed by DSM-IV) could be used to diagnose hypomania of BP-II. Diagnosing hypomania by counting a checklist of symptoms should make it easier to diagnose BP-II, and should reduce the current high misdiagnosis of BP-II as MDD, significantly impacting the treatment of depression. PMID- 17129710 TI - Temperament and character dimensions of the relatives of schizophrenia patients and controls: the relationship between schizotypal features and personality. AB - PURPOSE: Previous findings indicated that schizophrenia patients might have a different personality structure from the general population on several dimensions of temperament and character. Some authors proposed that HA might be a marker of underlying genetic vulnerability to schizophrenia. Studies on high-risk subjects and first degree relatives of patients is essential to test the value of a measure as a marker of genetic vulnerability to a disease. Few studies tested the biopsychosocial model of personality on unaffected relatives of schizophrenia. SUBJECTS AND METHODS: We compared the Temperament and Character (TCI) profiles of 94 first degree relatives of schizophrenia and 75 controls. We also investigated the relationship between schizotypy and TCI dimensions in the study sample. RESULTS: The harm avoidance scores of the relatives of schizophrenia patients with schizotypal features were significantly higher. Self transcendence scores were also significantly higher among relatives with schizotypal features. In contrast, the relatives of the patients with schizophrenia who did not have schizotypal features had higher SD and C scores than the control group. DISCUSSION AND CONCLUSION: This finding is consistent with the previous findings which suggested harm avoidance as a vulnerability indicator of schizophrenia. Some character features like self transcendence might be also associated with schizotypal features. PMID- 17129711 TI - Outcome and its predictors in schizophrenia within the Northern Finland 1966 Birth Cohort. AB - PURPOSE: We report clinical and social outcomes of schizophrenia in the longitudinal, population-based Northern Finland 1966 Birth Cohort, and describe associated demographic, developmental and illness-related factors. SUBJECTS AND METHODS: Subjects with DSM-III-R schizophrenia (n=59) were followed prospectively from mid-gestation up to age 35 years. Outcome measures included positive and negative symptoms, psychiatric hospitalisations, social and occupational functioning. Several definitions of good and poor outcome were explored, and developmental, socio-demographic and clinical predictors of outcomes were analysed. RESULTS: Good clinical outcome varied from 10% to 59%, and good social outcome 15-46%, depending on definition. Poor clinical outcome varied 41-77% and poor social 37-54%. Lack of friends in childhood, father's high social class, lower school performance and earlier age of illness onset predicted poor outcomes. DISCUSSION: The outcomes of schizophrenia in this study depended on definitions used but were relatively poor. The age of illness onset, father's social class, school performance and poor social contacts in childhood were only statistically significant predictors. CONCLUSION: Definitions of outcome have a major effect on estimates for proportions of good and bad outcomes and on the predictors of outcomes. However, regardless of which definitions were used, the outcome of schizophrenia in this population-based sample was generally bleak. PMID- 17129712 TI - Schizophrenic subtype, seasonality of birth and social class: a preliminary analysis. AB - OBJECTIVE: The neurodevelopmental model of schizophrenia includes the etiological impact of fetal brain stressors possibly connected with birth seasonality. Specification of social class of origin (SES) as a related risk factor remains unexamined as does type of schizophrenia as an outcome variable. The objective of this study was to test for an interconnection between SES, type of schizophrenia and seasonality of birth. METHODS: Patients (N=436) from a United States psychiatric hospital were separated into deficit/non-deficit presentation and bifurcated into poor/non-poor SES. Birth seasonality was assessed by months hypothetically connected with winter-related trimesters of gestation. RESULTS: Results showed that there is a significant difference (p=0.0411) in the monthly birth patterns of poor vs. non-poor patients and that the difference connects with the likelihood of deficit vs. non-deficit schizophrenia. Specifically, an elevated proportion of patients with deficit schizophrenia were born to impoverished women who likely conceived in January. Findings were confirmed by multiple levels of statistical assessment including log linear analysis. CONCLUSION: The resultant model suggests the environmental location (lower SES) and timing (winter conception) of adult schizophrenia with poor outcome (deficit). PMID- 17129713 TI - Cut-off values of fasting plasma glucose for detecting impaired glucose tolerance in Japanese population--relation between fasting and 2-h plasma glucose. PMID- 17129714 TI - Fluorescein transport properties across artificial lipid membranes, Caco-2 cell monolayers and rat jejunum. AB - Membrane transport characteristics of a paracellular permeability marker fluorescein were evaluated using artificial membrane, Caco-2 cell monolayers and rat jejunum, all mounted in side-by-side diffusion cells. Modified Ringer buffers with varied pH values were applied as incubation salines on both sides of artificial membrane, cell culture monolayers or rat jejunum. Passive transport according to pH partition theory was determined using all three permeability models. In addition to that, active transport of fluorescein in the M-S (mucosal to-serosal) direction through rat jejunum was observed. The highest M-S P(app) values regarding the active transport through the rat jejunum were observed in incubation saline with pH 6.5. Fluorescein transport through the rat jejunum was inhibited by DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid) and alpha CHC (alpha-cyano-4-hydroxycinnamic acid). Thus, we assume that two pH-dependent influx transporters could be involved in the fluorescein membrane transport through the intestinal (jejunal) epithelium. One is very likely an MCT (monocarboxylic acid cotransporter) isoform, inhibited by specific MCT inhibitor alpha-CHC, while the involvement of the second one with overlapping substrate/inhibitor specificities (most probably a member of the organic anion transporting polypeptide family, inhibited at least partially by DIDS) could not be excluded. PMID- 17129715 TI - Fibrin glue system for adjuvant brachytherapy of brain tumors with 188Re and 186Re-labeled microspheres. AB - Brain tumors such as glioblastoma reappear in their original location in almost 50% of cases. To prevent this recurrence, we developed a radiopharmaceutical system that consists of a gel applied immediately after surgical resection of a brain tumor to deliver local radiation booster doses. The gel, which strongly adheres to tissue in the treatment area, consists of fibrin glue containing the beta-emitters rhenium-188 and rhenium-186 in microsphere-bound form. Such microspheres can be prepared by short (2 h or less) neutron activation even in low neutron flux reactors, yielding a mixture of the two beta-emitters rhenium 188 (E(max)=2.1 MeV, half life=17 h) and rhenium-186 (E(max)=1.1 MeV, half life=90.6h). The dosimetry of this rhenium-188/rhenium-186 fibrin glue system was determined using gafchromic film measurements. The treatment efficacy of the radioactive fibrin glue was measured in a 9L-glioblastoma rat model. All animals receiving the non-radioactive fibrin glue died within 17+/-3 days, whereas 60% of the treated animals survived 36 days, the final length of the experiment. Control animals that were treated with the same amount of radioactive fibrin glue, but had not received a previous tumor cell injection, showed no toxic effects over one year. The beta-radiation emitting rhenium-188/rhenium-186-based gel thus provides an effective method of delivering high doses of local radiation to tumor tissue, particularly to wet areas where high adhesive strength and long-term radiation (with or without drug) delivery are needed. PMID- 17129717 TI - Domains 17-27 of tropoelastin contain key regions of contact for coacervation and contain an unusual turn-containing crosslinking domain. AB - The central region of tropoelastin including domains 19-25 of human tropoelastin forms a hot-spot for contacts during the inter-molecular association of tropoelastin by coacervation [Wise, S.G., Mithieux, S.M., Raftery, M.J. and Weiss, A.S (2005). "Specificity in the coacervation of tropoelastin: solvent exposed lysines." Journal of Structural Biology 149: 273-81.]. We explored the physical properties of this central region using a sub-fragment bordered by domains 17-27 of human tropoelastin (SHEL 17-27) and identified the intra- and inter-molecular contacts it forms during coacervation. A homobifunctional amine reactive crosslinker (with a maximum reach of 11 A, corresponding to approximately 7 residues in an extended polypeptide chain) was used to capture these contacts and crosslinked regions were identified after protease cleavage and mass spectrometry (MS) with MS/MS verification. An intermolecular crosslink formed between the lysines at positions 353 of each strand of tropoelastin at the lowest of crosslinker concentrations and was observed in all samples tested, suggesting that this residue forms an important initial contact during coacervation. At higher crosslinker concentrations, residues K425 and K437 showed the highest levels of involvement in crosslinks. An intramolecular crosslink between these K425 and K437, separated by 11 residues, indicated that a structural bend must serve to bring these residues into close proximity. These studies were complemented by small angle X-ray scattering studies that confirmed a bend in this important subfragment of the tropoelastin molecule. PMID- 17129718 TI - Expression and function of Toll-like receptors in T lymphocytes. AB - Toll-like receptors (TLRs) are widely expressed in the innate immune system. They recognize conserved microbial ligands such as bacterial lipopolysaccharide, lipopeptides or viral and bacterial RNA and DNA. TLRs play an essential role in innate immune responses and in the initiation of adaptive immune responses. However, certain TLRs are also expressed in T lymphocytes, and the respective ligands can directly modulate T cell function. TLR2, TLR3, TLR5 and TLR9 act as co-stimulatory receptors to enhance proliferation and/or cytokine production of T cell receptor-stimulated T lymphocytes. In addition, TLR2, TLR5 and TLR8 modulate the suppressive activity of naturally occurring CD25(+)CD4(+) regulatory T cells. The direct responsiveness of T lymphocytes to TLR ligands offers new perspectives for the immunotherapeutic manipulation of T cell responses. PMID- 17129719 TI - Autophagy in MHC class II antigen processing. AB - Durable adaptive immunity is dependent upon CD4 T-cell recognition of MHC class II molecules that display peptides from exogenous and endogenous antigens. Endogenously expressed cytosolic and nuclear antigens access MHC class II by way of several intracellular autophagic routes. These pathways include macroautophagy, microautophagy and chaperone-mediated autophagy. Macroautophagy can deliver antigens into autophagosomes for processing by acidic proteases before MHC class II presentation. However, other endogenous antigens are processed by cytoplasmic proteases, yielding fragments that translocate via chaperone-mediated autophagy into the endosomal network to intersect MHC class II. Cross-talk between autophagy pathways, particularly in response to stress, appears to balance the relative efficiency of each pathway. This might limit redundancy, giving MHC class II broader access to antigens within intracellular compartments distinct from the endosomal network. PMID- 17129720 TI - Rabbit allergens: a significant risk for allergic sensitization in subjects without occupational exposure. AB - BACKGROUND: Although rabbits are becoming popular as pets, data about the characteristics of allergic sensitization to rabbit allergens in patients without professional exposure are scarce. AIMS OF THE STUDY: To determine the characteristics of allergic sensitization to pet rabbits, and the role of direct and indirect exposure to rabbits and rabbit allergens in non-professionally exposed patients. METHODS: From among 1124 consecutive outpatients, we selected all subjects with an immediate skin reaction to rabbit dander. A clinical history including a careful evaluation of the modality of rabbit exposure, the results of skin-prick tests (SPTs) and total/specific IgE antibodies were recorded. The prevalence of rabbit ownership in the Naples area was also calculated. RESULTS: Among 753 SPT-positive patients, 20 (2.65%) were sensitized to rabbit dander (5 patients were mono-sensitized). Fifteen patients reported direct rabbit contact (7 were rabbit owners and 8 had occasional contact outside the home); 3 patients had indirect exposure through contact with rabbit owners and 2 patients denied any direct or indirect exposure. Rabbit mono-sensitized owners of pet rabbits had persistent (moderate-severe) symptoms. The prevalence of rabbit ownership is 1.56%. CONCLUSIONS: In susceptible not professionally exposed individuals, direct rabbit contact and, in some cases, indirect or no apparent exposure, may induce allergic sensitization to rabbit allergens. A progressive increase in rabbit sensitization (also by indirect exposure) may be expected as a consequence of the increase in rabbit ownership. PMID- 17129721 TI - Electrical detection of DNA hybridization: three extraction techniques based on interdigitated Al/Al2O3 capacitors. AB - Based on interdigitated aluminum electrodes covered with Al(2)O(3) and silver precipitation via biotin-antibody coupled gold nano-labels as signal enhancement, three complementary electrical methods were used and compared to detect the hybridization of target DNA for concentrations down to the 50 pM of a PCR product from cytochrome P450 2b2 gene. Human hepatic cytochrome P450 (CYP) enzymes participate in detoxification metabolism of xenobiotics. Therefore, determination of mutational status of P450 gene in a patient could have a significant impact on the choice of a medical treatment. Our three electrical extraction procedures are performed on the same interdigitated capacitive sensor lying on a passivated silicon substrate and consist in the measurement of respectively the low frequency inter-electrodes capacitance, the high-frequency self-resonance frequency, and the equivalent MOS capacitance between the short-circuited electrodes and the backside metallization of the silicon substrate. This study is the first of its kind as it opens the way for correlation studies and noise reduction techniques based on multiple electrical measurements of the same DNA hybridization event with a single sensor. PMID- 17129722 TI - Direct electron transfer with yeast cells and construction of a mediatorless microbial fuel cell. AB - The direct electron transfer exhibited by the yeast cells, Hansenula anomala has been demonstrated using the electrochemical technique cyclic voltammetry by immobilizing the microorganisms by two different methods viz., physical adsorption and covalent linkage. The analysis of redox enzymes present in the outer membrane of the microorganisms has been carried out in this work. This paper demonstrates that yeast cells with redox enzymes present in their outer membrane are capable of communicating directly with the electrode surface and contribute to current generation in a mediatorless biofuel cells. The efficiency of current generation has been evaluated using three anode materials. PMID- 17129723 TI - Profiling of molecular interactions in real time using acoustic detection. AB - Acoustic sensors that exploit resonating quartz crystals to directly detect the binding of an analyte to a receptor are finding increasing utility in the quantification of clinically relevant analytes. We have developed a novel acoustic detection technology, which we term resonant acoustic profiling (RAP). This technology builds on the fundamental basics of the "quartz crystal microbalance" or "QCM" with several key additional features including two- or four-channel automated sample delivery, in-line referencing and microfluidic sensor 'cassettes' that are pre-coated with easy-to-use surface chemistries. Example applications are described for the quantification of myoglobin concentration and its interaction kinetics, and for the ranking of enzyme cofactor specificities. PMID- 17129724 TI - Design and synthesis of furoxan-based nitric oxide-releasing glucocorticoid derivatives with potent anti-inflammatory activity and improved safety. AB - A series of furoxan-based nitric oxide-releasing glucocorticoid derivatives was synthesized. The pharmacological assays indicated that three compounds, including I(4), I(5), and I(6), had anti-inflammatory activity. Furthermore compared with the leading compound hydrocortisone the safety of I(6) was greatly improved. Due to releasing NO in vivo the side effects of glucocorticoids, including hypertension and osteoporosis, were effectively avoided. PMID- 17129725 TI - Design and synthesis of a novel class of dual PPARgamma/delta agonists. AB - The design and synthesis of dual PPAR gamma/delta agonist (R)-3-{2-ethyl-4-[3-(4 ethyl-2-pyridin-2-yl-phenoxy)-butoxy]-phenyl}propionic acid is described. This compound dose-dependently lowered plasma glucose in hyperglycemic male Zucker diabetic fatty (ZDF) rats and produced less weight gain relative to rosiglitazone at an equivalent level of glucose control. PMID- 17129726 TI - 3,4-Dihydro-2H-benzoxazinones as dual-acting 5-HT1A receptor antagonists and serotonin reuptake inhibitors. AB - Investigation of halogen substitution in lead compound 1 has led to the identification of analogues which combine high affinity for 5-HT(1A) receptors and potent serotonin reuptake inhibitory activity. Several compounds show an improved selectivity over 5-HT(1B) and 5-HT(1D) receptors and a superior pharmacokinetic profile in the rat. PMID- 17129727 TI - Painful enlargement of the calf muscles in limb girdle muscular dystrophy type 2B (LGMD2B) with a novel compound heterozygous mutation in DYSF. AB - Limb girdle muscular dystrophy type 2B (LGMD2B) and Miyoshi Myopathy are caused by mutations in the dysferlin gene. The phenotype of these allelic disease variants can vary considerably. We report on an adolescent female with a severe and rapidly progressing clinical course of LGMD2B which has been suggested by the muscle histopathology and Western blot and proven by mutation analysis in the Dysferlin gene. We detected a novel compound heterozygous mutation of which one affects the extracellular part of the protein. This is the first report on a mutation in this region of dysferlin and might explain the unusual phenotype of the patient. PMID- 17129728 TI - The vascular niche and its basement membrane. AB - Over the past few years, scientists have realized that many cellular and developmental processes, including pancreatic beta-cell growth and differentiation, stem cell and progenitor cell proliferation and cancer cell metastasis, occur in what are known as 'vascular niches'. Despite increasing numbers of reports on these niches, few common mechanisms have been identified to explain their various effects. Here, we define the term 'vascular niche' and suggest that a common and conserved feature of this niche is to provide a basement membrane to cells that are unable to form their own. We further propose that these cells require a vascular niche when they retain a high degree of plasticity. PMID- 17129729 TI - Contribution of calf muscle-tendon properties to single-leg stance ability in the absence of visual feedback in relation to ageing. AB - We tested the hypothesis that the importance of calf muscle-tendon properties for maintaining balance during single-leg stance increases in the absence of visual feedback. Trial duration, centre of pressure displacement normalized for trial duration (nD), electromyographic (EMG) activity of the main ankle plantarflexors and dorsiflexors, and ground reaction forces (F(P)), were measured in 20 younger (aged 18+/-1 years; mean+/-S.E.M.) and 28 older (aged 68+/-1 years) healthy participants during single-leg stance in eyes-open (EO) and eyes-closed (EC) conditions. Plantarflexor muscle strength, activation capacity and tendon stiffness were assessed by dynamometry, electrical stimulation and ultrasonography, respectively. Muscle-tendon characteristics in the older participants were up to 55% (P<0.0001) lower compared with their younger counterparts. Trial duration, F(P), nD and EMG changed in EC compared with EO by 21% and up to approximately 4.6 times (P<0.01) in the two population groups. Multiple linear regression with age and the three muscle-tendon properties showed a substantial increment in EC compared to EO for trial duration (R(2)=0.86 versus R(2)=0.72), but a similarity for nD (R(2)=0.36 versus R(2)=0.33). These results suggest that factors other than the ones that we examined become important when steadiness rather than stance duration is the object of single-leg stance in the absence of vision. PMID- 17129730 TI - Preparing the way: fungal motors in microtubule organization. AB - Fungal growth, development and pathogenicity require hyphal tip growth, which is supported by polar exocytosis at the expanding growth region. It is assumed that molecular motors transport growth supplies along the fibrous elements of the cytoskeleton, such as microtubules, to the hyphal apex. Recent advances in live cell imaging of fungi revealed additional roles for motors in organizing their own tracks. These unexpected roles of the molecular motors are modifying microtubule dynamics directly, targeting stability-determining factors to microtubule plus ends, and transporting and arranging already-assembled microtubules. PMID- 17129731 TI - A prospective study of 3-day versus 2-week immobilization period after anterior cruciate ligament reconstruction. AB - BACKGROUND: Aggressive early rehabilitation is recommended after reconstruction of the anterior cruciate ligament (ACL) using a patellar tendon-bone graft. However, rehabilitation after reconstruction of ACL using the multistranded hamstring is controversial. In this study, we compared the clinical results of 3 day immobilization after reconstruction of ACL using the multistranded hamstring, with those of 2-week immobilization prospectively. METHODS: A total of 30 consecutive patients with ruptured ACL underwent single-socket ACL reconstruction. 10 male and 5 female patients (mean age of 29.2+/-10.0 years) were prospectively randomized for a 3-day immobilization period, and 6 male and 9 female patients (mean age of 27.3+/-10.9 years) were randomized for a 2-week immobilization period. We measured the anterior laxity with a KT-2000 arthrometer, joint position sense, and thigh muscle strength at 3, 6 and 12 months after surgery. RESULTS AND CONCLUSIONS: There were no statistically significant differences between the two groups. We concluded that the post operative 3-day immobilization period is preferable to the longer 2-week period in our rehabilitation program from the viewpoint of psychological load to the patients and the duration of hospitalization after ACL reconstruction using multistranded hamstring tendons. PMID- 17129732 TI - Chemoenzymatic synthesis of artificial glycopolypeptides containing multivalent sialyloligosaccharides with a gamma-polyglutamic acid backbone and their effect on inhibition of infection by influenza viruses. AB - Highly water-soluble, artificial glycopolypeptides with a gamma-polyglutamic acid (gamma-PGA) backbone derived from Bacillus subtilis sp. and multivalent sialyloligosaccharide units have been chemoenzymatically synthesized as potential polymeric inhibitors of infection by bird and human influenza viruses. 5 Trifluoroacetamidopentyl beta-N-acetyllactosaminide and 5 trifluoroacetamidopentyl beta-lactoside were enzymatically synthesized from LacNAc and lactose, respectively, by cellulase-mediated condensation with 5 trifluoroacetamido-1-pentanol. After deacetylation, the resulting 5-aminopentyl beta-LacNAc and beta-lactoside glycosides were coupled to the alpha-carboxyl groups of the gamma-PGA side chains. The artificial glycopolypeptides carrying LacNAc and lactose were further converted to Neu5Acalpha2-(3/6)Galbeta1-4Glcbeta and Neu5Acalpha2-(3/6)Galbeta1-4GlcNAcbeta sialyloligosaccharide units by alpha2,3- and alpha2,6-sialyltransferase, respectively. The interaction of these glycopolypeptides with various influenza virus strains has been investigated by three different methods. Glycopolypeptides carrying Neu5Acalpha2,6LacNAc inhibited hemagglutination mediated by influenza A and B viruses, and their relative binding affinities for hemagglutinin were 10(2)- to 10(4)-fold higher than that of the naturally occurring fetuin control. A glycopolypeptide carrying Neu5Acalpha2,6LacNAc inhibited infection by A/Memphis/1/71 (H3N2) 93 times more strongly than fetuin, as assessed by cytopathic effects on virus-infected MDCK cells. The avian virus [A/duck/Hong kong/4/78 (H5N3)] bound strongly to Neu5Acalpha2,3LacNAc/Lac-carrying glycopolypeptides, whereas the human virus [A/Memphis/1/71 (H3N2)] bound to Neu5Acalpha2,6LacNAc in preference to Neu5Acalpha2,6Lac. Taken together, these results indicate that the binding of viruses to terminal sialic acids is markedly affected by the structure of the asialo portion, in this case either LacNAc or lactose, in the sugar chain of glycopolypeptides. PMID- 17129733 TI - Novel antitumor agent family of 1H-benzo[c,d]indol-2-one with flexible basic side chains: synthesis and biological evaluation. AB - A series of mono-1H-benzo[c,d]indol-2-one with different amine side chains and bis-1H-benzo[c,d]indol-2-one as novel family of DNA intercalators were designed and synthesized, the contributions of aromatic chromophores and amine side chains for DNA binding properties, for example, intercalation and electrostatic binding, respectively, were evaluated. Among them, A3 tailed with N,N-dimethylamino-ethyl ethane-1,2-diamine showed selective anti-tumor activities against cell lines A549 and P388 with IC(50) 0.428microm and 1.69microm. PMID- 17129734 TI - Gender differences in drug responses. AB - This review summarizes gender differences (GDs) in drug response. Although GDs have been described both in pharmacodynamics and pharmacokinetics, their role in clinical practice is not yet completely elucidated. The evidence that women have been less enrolled in clinical trials and that a gender-specific analysis usually is not included in the evaluation of results, contributes largely to this uncertainty. Consequently, adverse drug reactions (ADRs) are still higher in females than in males. Since sex is a fundamental biological variable that cannot be discounted, GDs in pharmacology have to be considered in order to improve drug safety efficacy and to optimize medical therapy both in men and women. PMID- 17129735 TI - Efficient expression of secreted proteases via recombinant BacMam virus. AB - Baculovirus vectors engineered to contain mammalian cell-active promoter elements have been described as an efficient method for transduction of a broad spectrum of human cell lines at high frequency. In the first large-scale comparative study of secreted protein production using these viral vectors, we have evaluated production of 16 recombinant enzymes--specifically, we exploited these viral vectors, termed 'BacMam' viruses, to drive expression of a panel of proteases selected from all four major mechanistic classes, including secreted, lysosomal, endosomal, and type I transmembrane proteins. To allow a generic purification strategy, coding sequences were truncated to remove transmembrane and/or subcellular retention signals before introduction, in parallel, into a C terminally Fc-tagged BacMam transfer vector. BacMam viruses were generated and subsequently evaluated for expression of Fc-tagged protein in virus-transduced HEK-F cells. The common Fc-tag enabled single-step affinity purification of secreted recombinant protein from the culture medium. Yields were excellent, with 14 of 16 genes expressed producing 10-30 mg or more purified protein per litre of culture using standardised transduction conditions. At this level, reagent demands for a typical protease high-throughput screen (HTS) could be met from expression cultures as small as 0.1-0.5 L. Our results indicate this expression system offers a highly efficient and scaleable method for production of enzymatically-active secreted proteases and may therefore represent a novel method of protein production for other secreted enzymes with significant advantages over the diverse approaches in current use. PMID- 17129736 TI - Local refinement: an attempt to correct for shrinkage and distortion in electron tomography. AB - A critical problem in electron tomography is the deformation of the specimen due to radiation, or "shrinkage," which interferes with image alignment and thereby limits resolution. Here, we describe a general strategy for refining preliminary reconstructions which allows the damage due to the shrinkage of plastic-embedded thin sectioned specimens (50-80 nm) to be corrected. The basic steps of the strategy involve: (a) the partition of the preliminary reconstruction into sub volumes; (b) the extraction of corresponding sub-areas for each sub-volume from the micrographs of the tilt series; (c) the re-projection of each sub-volume according to the orientation parameters; and (d) the refinement of these parameters by correlating each sub-area to the corresponding computed projection. We tested the strategy by refining chemical synapses reconstructed from series imaged with conical, double and single tilt geometries. The results gathered with local refinement were evaluated by visually inspecting the structure of biological membranes in the maps. In an effort to quantify these improvements, we studied the refined maps using correlation criteria and mapped the corrections applied to the orientation parameters in each sub-volume of the reconstruction. Simulation experiments complemented the data gathered by correlation analysis. Based on these criteria, we concluded that local refinement significantly improves the overall quality of the reconstructions of chemical synapses calculated from series imaged with conical and double tilt geometries. PMID- 17129737 TI - Reticulate evolution in the Acrolophus subgroup (Centaurea L., Compositae) from the western Mediterranean: origin and diversification of section Willkommia Blanca. AB - Section Willkommia (Centaurea, Compositae) is endemic to the east-central portion of the Iberian Peninsula and northwestern Africa. The section has been included with sections Acrolophus and Phalolepis in the informal subgroup Acrolophus. We have used a molecular phylogenetic approach to test the hypothesis proposed by earlier authors that the diversification of section Willkommia involved a schizoendemic process from an ancestral syngameon. Comparative analysis of the transcribed spacer sequences of the nuclear ribosomal DNA (ITS and 3'ETS) regions reveals the presence of three different types of ribosomal sequences in the Acrolophus subgroup (referred to here as the Willkommia, Acrolophus-Phalolepis and Simulans ribotypes) which show a sectional-independent geographic structure. This evidence, together with the presence of additive polymorphic sites in the Willkommia sequences which agree with the geographic distribution of the taxa, suggests that members of section Willkommia and the western Mediterranean taxa of sections Acrolophus and Phalolepis fit a reticulate evolution model. PMID- 17129738 TI - A mitochondrial-targeting signal is present in the non-catalytic domain of the MELK protein kinase. AB - MELK is a cell cycle-regulated protein kinase involved in cell cycle progression, proliferation, tumor growth and mRNA splicing. MELK is localized in the cytoplasm and the nucleus during interphase and at the cell cortex during anaphase and telophase. In this report, we show that the regulatory domain of Xenopus MELK when tagged at its C-terminus with the green fluorescent protein (GFP), co localizes with mitochondria in Xenopus XL2 cells. Significantly, the presence of a mitochondrial targeting signal at the N-terminus of this fusion protein was predicted by bioinformatics analyses. In agreement with previous reports on mitochondrial proteins, placing the GFP at the N-terminus inhibited the mitochondrial targeting of the MELK fragment and, furthermore, the regulatory domain without a tag co-localizes with mitochondria. These results demonstrate the presence of a mitochondrial targeting signal at the N-terminus of the MC domain of MELK. This mitochondrial targeting signal was also functional in human HeLa cells. PMID- 17129739 TI - Hippocampal long-term potentiation, memory, and longevity in mice that overexpress mitochondrial superoxide dismutase. AB - Superoxide has been shown to be critically involved in several pathological manifestations of aging animals. In contrast, superoxide also can act as a signaling molecule to modulate signal transduction cascades required for hippocampal synaptic plasticity. Mitochondrial superoxide dismutase (SOD-2 or Mn SOD) is a key antioxidant enzyme that scavenges superoxide. Thus, SOD-2 may not only prevent aging-related oxidative stress, but may also regulate redox signaling in young animals. We used transgenic mice overexpressing SOD-2 to study the role of mitochondrial superoxide in aging, synaptic plasticity, and memory associated behavior. We found that overexpression of SOD-2 had no obvious effect on synaptic plasticity and memory formation in young mice, and could not rescue the age-related impairments in either synaptic plasticity or memory in old mice. However, SOD-2 overexpression did decrease mitochondrial superoxide in hippocampal neurons, and extended the lifespan of the mice. These findings increase our knowledge of the role of mitochondrial superoxide in physiological and pathological processes in the brain. PMID- 17129740 TI - Osmo and ionic regulation of black tiger prawn (Penaeus monodon Fabricius 1798) juveniles exposed to K(+) deficient inland saline water at different salinities. AB - An 11-day trial was conducted to investigate the osmoregulatory capacity (OC) and regulation of K(+), Na(+), Ca(2+) and Mg(2+) of Penaeus monodon juveniles when exposed to K(+) deficient inland saline water (ISW) of four different salinities (5, 15, 25 and 35 ppt). The survival of juveniles showed a positive linear relationship (R(2) ranging from 0.72 to 0.98) with salinity. At the end of the trial, juveniles were able to survive only in 5 ppt of ISW and showed no changes in OC when transferred from ocean water (OW) to ISW. Further, the OC of juveniles in 5 ppt of ISW was significantly different (P<0.05) from the OC of juveniles exposed to 15, 25 and 35 ppt and exhibited strong serum K(+), Na(+), Ca(2+) and Mg(2+) regulation monitored over 16 h. In contrast, at 35 ppt, significant decrease (P<0.05) in serum K(+) and Mg(2+) concentrations and accumulation of serum Na(+) concentration occurred after 16 h of exposure to ISW. At higher salinity, an increase in serum Na(+) concentration leads to an increase in the serum osmolality of the juveniles, which in turn causes decrease in the OC of the juveniles. The results of this study suggest that K(+) deficiency in ISW has a negative effect on survival, OC and the ability of P. monodon juveniles to regulate serum Na(+), K(+), Ca(2+) and Mg(2+) concentrations. These effects are compounded as salinity increases. PMID- 17129741 TI - Relationships between the functional PPARalpha Leu162Val polymorphism and obesity, type 2 diabetes, dyslipidaemia, and related quantitative traits in studies of 5799 middle-aged white people. AB - Peroxisome proliferator-activated receptor-alpha (PPARalpha) is a nuclear receptor capable of regulating the expression of genes involved in peroxisomal and mitochondrial beta-oxidation pathways. The common Leu162Val polymorphism in the gene encoding PPARalpha has inconsistently shown association with quantitative traits related to obesity, type 2 diabetes, and dyslipidaemia. We genotyped the Leu162Val polymorphism in 1383 patients with type 2 diabetes and 4401 control subjects with normal glucose tolerance (NGT) without showing any association between diabetes and genotype. In addition, the Leu162Val polymorphism was not associated with WHO-defined obesity or dyslipidaemia in case control settings involving 961 obese and 2563 lean subjects and 1399 dyslipidaemic and 4399 normolipidaemic subjects, respectively. Quantitative trait studies of metabolic variables were carried out in 5799 middle-aged, treatment naive subjects showing a difference in fasting serum triglyceride concentrations among homozygous Val-carriers (Leu/Leu+Leu/Val, n=5782, 1.33+/-1.35 mmol/l vs. Val/Val, n=17, 2.22+/-2.4 mmol/l, p=0.007). Similarly, Val/Val was associated with increased fasting serum total cholesterol concentrations (p=0.01). In conclusion, in a relative large-scale study of middle-aged whites we found no evidence of association between the PPARalpha Leu162Val polymorphism and obesity or type 2 diabetes. If replicated, the Val162Val variant may, however, confer an increase in fasting levels of serum lipids. PMID- 17129743 TI - Partial hypoxanthine-guanine phosphoribosyl transferase deficiency without elevated urinary hypoxanthine excretion. AB - Partial hypoxanthine-guanine phosphoribosyl transferase (HGPRT) deficiency, also known as the Kelley-Seegmiller syndrome, can give rise to a wide range of neurological symptoms, and renal insufficiency. Biochemically, it is characterized by high uric acid concentrations in blood, high uric acid and hypoxanthine excretion in urine, and decreased activity of hypoxanthine-guanine phosphoribosyl transferase activity (HGPRT). However, normal uric acid concentrations in blood and uric acid excretions in urine have been reported. Here, a boy is presented with normal development and suffering from recurrent attacks of acute renal failure with slightly to clearly increased urinary uric acid excretion. Between these attacks, episodes of elevated urinary excretion of uric acid were observed with normal blood concentrations of uric acid and normal urinary excretion of hypoxanthine. HGPRT activity in erythrocytes, leukocytes, and fibroblasts was found to be strongly decreased. This case shows that not only normal blood uric acid but also normal urinary hypoxanthine concentrations do not exclude the diagnosis of partial HGPRT deficiency. PMID- 17129744 TI - Recognition memory for unfamiliar faces in Parkinson's disease: behavioral and electrophysiologic measures. AB - We analyzed event-related potentials (ERPs) and behavioral measurements during a recognition memory task in 15 normal elderly subjects and 15 patients with Parkinson's disease (PD). To elicit ERPs unfamiliar faces were repeated immediately after initial presentation (at lag 0), after one intervening face (at lag 1) or at lag 3. Compared to normal controls, PD patients showed decreased accuracy in recognizing new unfamiliar faces. P170 latency and amplitude were similar between both groups. ERP amplitude between 300 and 500 ms after the stimulus in control subjects showed a positive shift (ERP repetition effect) for lag 0 at all sites and for lag 1 and 3 repetitions at the Fz site, while effects in the PD group were not noted at any site, even for the lag 0 repetition. ERP waveforms for the first presentation of faces in PD patients showed a significant positive shift compared to normal controls. These data suggest intact perception but impaired recognition memory for unfamiliar faces in PD. In addition, recognition memory deficits in PD may result from impairment of comparison of structural representations of presented faces with stored representations of faces known to the observer. PMID- 17129742 TI - Multiple mechanisms of growth hormone-regulated gene transcription. AB - Diverse physiological actions of growth hormone (GH) are mediated by changes in gene transcription. Transcription can be regulated at several levels, including post-translational modification of transcription factors, and formation of multiprotein complexes involving transcription factors, co-regulators and additional nuclear proteins; these serve as targets for regulation by hormones and signaling pathways. Evidence that GH regulates transcription at multiple levels is exemplified by analysis of the proto-oncogene c-fos. Among the GH regulated transcription factors on c-fos, C/EBPbeta appears to be key, since depletion of C/EBPbeta by RNA interference blocks the stimulation of c-fos by GH. The phosphorylation state of C/EBPbeta and its ability to activate transcription are regulated by GH through MAPK and PI3K/Akt-mediated signaling cascades. The acetylation of C/EBPbeta also contributes to its ability to activate c-fos transcription. These and other post-translational modifications of C/EBPbeta appear to be integrated for regulation of transcription by GH. The formation of nuclear proteins into complexes associated with DNA-bound transcription factors is also regulated by GH. Both C/EBPbeta and the co-activator p300 are recruited to c-fos in response to GH, altering c-fos promoter activation. In addition, GH rapidly induces spatio-temporal re-localization of C/EBPbeta within the nucleus. Thus, GH-regulated gene transcription mediated by C/EBPbeta reflects the integration of diverse mechanisms including post-translational modifications, modulation of protein complexes associated with DNA and re-localization of gene regulatory proteins. Similar integration involving other transcription factors, including Stats, appears to be a feature of regulation by GH of other gene targets. PMID- 17129745 TI - Keratinocyte serum-free medium maintains long-term liver gene expression and function in cultured rat hepatocytes by preventing the loss of liver-enriched transcription factors. AB - Freshly isolated hepatocytes rapidly lose their differentiated properties when placed in culture. Therefore, production of a simple culture system for maintaining the phenotype of hepatocytes in culture would greatly facilitate their study. Our aim was to identify conditions that could maintain the differentiated properties of hepatocytes for up to 28 days of culture. Adult rat hepatocytes were isolated and attached in Williams' medium E containing 10% serum. The medium was changed to either fresh Williams' medium E or keratinocyte serum-free medium supplemented with dexamethasone, epidermal growth factor and pituitary gland extract. The hepatic phenotype was then analysed using RT-PCR, immunohistochemistry, Western blotting and assays of liver function. Cells cultured in keratinocyte serum-free medium supplemented with dexamethasone, epidermal growth factor and pituitary gland extract maintained their phenotype for 3-4 weeks, based on expression of liver proteins, ureagenesis and response to xenobiotics. In contrast, hepatocytes cultured in Williams' medium E rapidly lost the expression of liver proteins after 3 days. Cells cultured in keratinocyte serum-free medium supplemented with dexamethasone, epidermal growth factor and pituitary gland extract maintained their expression of liver-enriched transcription factors (C/EBPalpha and beta, HNF4alpha and RXRalpha) while expression was either lost or reduced in cells cultured in Williams' medium E. These results suggest that keratinocyte serum-free medium supplemented with dexamethasone, epidermal growth factor and pituitary gland extract can maintain the hepatic phenotype for a prolonged period and that this is probably related to the continued expression of the liver-enriched transcription factors. PMID- 17129746 TI - Can generic expertise explain special processing for faces? AB - Does face recognition involve face-specific cognitive and neural processes ('domain specificity') or do faces only seem special because people have had more experience of individuating them than they have of individuating members of other homogeneous object categories ('the expertise hypothesis')? Here, we summarize new data that test these hypotheses by assessing whether classic face-selective effects - holistic processing, recognition impairments in prosopagnosia and fusiform face area activation - remain face selective in comparison with objects of expertise. We argue that evidence strongly supports domain specificity rather than the expertise hypothesis. We conclude that the crucial social function of face recognition does not reflect merely a general practice phenomenon and that it might be supported by evolved mechanisms (visual or nonvisual) and/or a sensitive period in infancy. PMID- 17129747 TI - Using multi-voxel pattern analysis of fMRI data to interpret overlapping functional activations. PMID- 17129748 TI - Attention and consciousness: two distinct brain processes. AB - The close relationship between attention and consciousness has led many scholars to conflate these processes. This article summarizes psychophysical evidence, arguing that top-down attention and consciousness are distinct phenomena that need not occur together and that can be manipulated using distinct paradigms. Subjects can become conscious of an isolated object or the gist of a scene despite the near absence of top-down attention; conversely, subjects can attend to perceptually invisible objects. Furthermore, top-down attention and consciousness can have opposing effects. Such dissociations are easier to understand when the different functions of these two processes are considered. Untangling their tight relationship is necessary for the scientific elucidation of consciousness and its material substrate. PMID- 17129749 TI - Perception and misperception of bias in human judgment. AB - Human judgment and decision making is distorted by an array of cognitive, perceptual and motivational biases. Recent evidence suggests that people tend to recognize (and even overestimate) the operation of bias in human judgment - except when that bias is their own. Aside from the general motive to self enhance, two primary sources of this 'bias blind spot' have been identified. One involves people's heavy weighting of introspective evidence when assessing their own bias, despite the tendency for bias to occur nonconsciously. The other involves people's conviction that their perceptions directly reflect reality, and that those who see things differently are therefore biased. People's tendency to deny their own bias, even while recognizing bias in others, reveals a profound shortcoming in self-awareness, with important consequences for interpersonal and intergroup conflict. PMID- 17129750 TI - Adding up the effects of cultural experience on the brain. AB - How does the brain represent number and perform mathematical calculations? According to a recent and provocative study by Tang and colleagues, it depends on which language you learn. They found that the divergent linguistic and cultural experiences of native Chinese and native English speakers are associated with distinct patterns of brain activity during mathematical processing. Their results raise important questions about the cognitive and neural specificity of cultural influences on mathematical processes and the core nature of mathematical cognition. PMID- 17129751 TI - CLE peptide ligands and their roles in establishing meristems. AB - Research in the past decade revealed that peptide ligands, also called peptide hormones, play a crucial role in intercellular communication and defense response in plants. Recent studies demonstrated that a family of plant-specific genes, CLAVATA3 (CLV3)/ENDOSPERM SURROUNDING REGION (ESR) (CLE), which has at least 31 members in Arabidopsis genome, are able to generate extracellular peptides to regulate cell division and differentiation. A hydroxyl 12-amino acid peptide derived from the conserved CLE motif of CLV3 promotes cell differentiation, whereas another CLE-derived peptide suppresses the differentiation. These peptides probably interact with membrane-bound, leucine-rich repeat receptor-like kinases (LRR-RLKs) to execute the decision between cell proliferation and differentiation. PMID- 17129752 TI - Trichostrongylina (Nematoda) parasitic in Phyllotis sp. (Rodentia: Sigmodontinae) from Argentina, with description of three new species. AB - The Trichostrongylina parasitic in the leaf-eared mouse Phyllotis sp. (Sigmodontinae) from the Province of Catamarca, Argentina are studied. Three new species of Stilestrongylus Freitas, Lent and Almeida, 1937 (Heligmosomoidea: Heligmonellidae: Nippostrongylinae) are described. Stilestrongylus andalgala n. sp. is distinguished from the most closely related species S. moreli Diaw, 1976 parasitic in Auliscomys boliviensis from Bolivia, by the absence of a common trunk of right rays 2 and 3, by left ray 4 shorter than ray 5, and by the shape of the female tail, without a mucron. Stilestrongylus catamarca n. sp. is distinguished from all the other species in the genus by hypertrophied rays 2, larger than rays 3, and from the closely related species Stilestrongylus barusi Durette-Desset, 1971, parasitic in Sigmodontomys alfari from Colombia, by right ray 4 longer than ray 5 and left ray 4 shorter than ray 5, by a larger spicule length/body length ratio (22.6% vs. 7.2%), and by the presence of a mucron on the female tail. Stilestrongylus gracielae n. sp. most closely resembles Stilestrongylus azarai Durette-Desset and Sutton, 1985, parasitic in Akodon azarae and in Graomys griseoflavus, and Stilestrongylus franciscanus Digiani and Durette-Desset, 2003, also parasitic in G. griseoflavus, both from Argentina. These latter differ from the new species by the right lobe of the caudal bursa: in S. azarai rays 4, 5 and 6 arise at same level from their common trunk and in S. franciscanus right rays 4 and 5 diverge at their distal extremity. Lamanema chavezi Becklund, 1963 (Molineoidea: Molineinae), a parasite of South American camelids, is reported in rodents for the second time. PMID- 17129753 TI - Applications of diamond crystal ATR FTIR spectroscopy to the characterization of ambers. AB - Diamond crystal ATR FTIR spectroscopy is a rapid technique with virtually no sample preparation which requires small sample amounts and showed potential in the study of ambers. FTIR spectra of ambers present discriminating patterns and can be used to distinguish amber from immature resins as copal, to determine local or Baltic origin of archaeological ambers and to detect most of the falsifications encountered in the amber commercialisation. PMID- 17129754 TI - Low-temperature photoluminescence of p-nitroaniline and o-methyl-p-nitroaniline crystals. AB - Photoemission spectra of para-nitroaniline (p-NA) and ortho-methyl-para nitroaniline (MNA) single crystals measured between 5 and 250K revealed that the MNA crystals emit phosphorescence and fluorescence in contrast with the p-NA crystals which have only fluorescence. It is assumed that the fluorescence of p NA crystals at 5K originates from the doublet state of negatively charged polarons while the fluorescence of MNA crystals at 60K is due to trap states. The Stokes shift between absorption and emission onsets, measured at 5K, is much larger in the p-NA crystals than in the MNA and meta-nitroaniline (m-NA) crystals. This fact is rationalized by different geometrical changes caused by excitation. PMID- 17129755 TI - Electron paramagnetic resonance study on manganese doped tetrathioureacopper(I) chloride single crystal. AB - Electron paramagnetic resonance (EPR) spectrum of a NLO material, manganese doped tetrathioureacopper(I) chloride single crystal was investigated at room temperature. The spectra reveal that manganese exists in tetravalent state in the host lattice in interstitial position and the crystal becomes NLO material whereas the pure crystal does not show NLO property. The spin Hamiltonian parameters are evaluated as g=1.9872 and A=157G. The transmission spectrum and the second harmonic generation efficiency confirm that the manganese doped single crystal exhibit NLO property. PMID- 17129756 TI - Development of nickel-doped lithium formate as potential EPR dosimeter for low dose determination. AB - EPR dosimetry employing L-alpha-alanine has been in vogue during the past few years, due to its tissue equivalence and linear dose response. However, L-alpha alanine dosimetry has been improved during the past years, the sensitivity of this material is still too low for clinical applications. Polycrystalline lithium formate doped with NiCl2 was therefore examined for radiation response in the dose range of clinical interest (<5 Gy) using CW EPR and pulse EPR techniques. At equal and moderate settings of microwave power and modulation amplitude lithium formate doped with 1.6 wt% of NiCl2 was almost four times more sensitive compared to L-alpha-alanine, which is the most common EPR dosimeter standard. It was shown that the nickel-doped lithium formate has an excellent radiation response with a low limit of the measurable dose, and a linear dose response in the range 1-5 Gy. The relaxation and power saturation studies showed that high microwave power can be applied during measurements to improve the sensitivity of this material as an EPR dosimeter. These results show that lithium formate doped with Ni(II) exhibits promising properties required for further development of an EPR dosimeter in the dose range typical for clinical dosimetry. PMID- 17129757 TI - NMR structure determination of a synthetic analogue of bacillomycin Lc reveals the strategic role of L-Asn1 in the natural iturinic antibiotics. AB - Iturins are a group of antifungal produced by Bacillus subtilis. All are cyclic lipopeptides with seven alpha-amino acids of configuration LDDLLDL and one beta amino fatty acid. The bacillomycin L is a member of this family and its NMR structure was previously resolved using the sequence Asp-Tyr-Asn-Ser-Gln-Ser-Thr. In this work, we carefully examined the NMR spectra of this compound and detected an error in the sequence. In fact, Asp1 and Gln5 need to be changed into Asn1 and Glu5, which therefore makes it identical to bacillomycin Lc. As a consequence, it now appears that all iturinic peptides with antibiotic activity share the common beta-amino fatty acid 8-L-Asn1-D-Tyr2-D-Asn3 sequence. To better understand the conformational influence of the acidic residue L-Asp1, present, for example in the inactive iturin C, the NMR structure of the synthetic analogue SCP [cyclo (L Asp1-D-Tyr2-D-Asn3-L-Ser4-L-Gln5-D-Ser6-L-Thr7-beta-Ala8)] was determined and compared with bacillomycin Lc recalculated with the corrected sequence. In both cases, the conformers obtained were separated into two families of similar energy which essentially differ in the number and type of turns. A detailed analysis of both cyclopeptide structures is presented here. In addition, CD and FTIR spectra were performed and confirmed the conformational differences observed by NMR between both cyclopeptides. PMID- 17129758 TI - Human Papillomavirus testing for the management of low-grade cervical abnormalities in the UK--Influence of age and testing strategy. AB - BACKGROUND: The results of the UK pilot studies were encouraging with respect to the introduction of Human Papillomavirus (HPV) testing as a means to improve the management of low-grade cytological abnormalities. However, several important unresolved issues related to HPV triage remain, two of which are: what type of HPV test should be used and what age group should be targeted. OBJECTIVES: To perform an evaluation of two commercial HPV detection tests and to correlate disease persistence and clearance with age and HPV status by the two tests. STUDY DESIGN: We performed an evaluation of two commercial HPV tests in a cross sectional analysis of 322 cervical cytology specimens with low-grade abnormalities. A subset of these specimens were then examined longitudinally, in order to correlate disease persistence and clearance with age and HPV status by the two detection tests. RESULTS: The two tests performed similarly with respect to the longitudinal identification/prediction of high-grade cervical disease. Age did not appear to be a factor in determining which cases went on to manifest high grade disease within 3 years of a low-grade result (p=0.678). CONCLUSIONS: This study weakens the case for age-adjusted HPV triage of low-grade cervical abnormalities. PMID- 17129759 TI - Serological evidence for Borna disease virus infection in humans, wild rodents and other vertebrates in Finland. AB - BACKGROUND: Borna disease virus (BDV) can infect many vertebrate species, including humans. BDV infection may lead to meningoencephalomyelitis in animals. An association with human neuropsychiatric diseases has been reported, but the causal relationship between BDV and human disease remains unclear. OBJECTIVES AND STUDY DESIGN: To find out whether BDV is present in Finland and to look for a potential reservoir, we examined a large panel of blood samples from different vertebrate species with immunofluorescence assay. Samples from horses, cats, dogs, sheep, cattle, large predators, grouse, wild rodents and humans were included. Most positive results were confirmed by other specific methods and in other laboratories. RESULTS AND CONCLUSIONS: BDV-specific antibodies were detected in 10 horses, 2 cats, as well as 2 horses and 1 dog from farms housing a previously detected seropositive horse. Interestingly, BDV-specific antibodies were further detected in three wild rodents. In humans, BDV-specific antibodies were detected in a veterinarian and in two patients suspected to have a Puumala hantavirus infection. Our serological analysis suggests that BDV infects various vertebrates in Finland, including humans. Furthermore, our data indicate for the first time that BDV infects also wild rodents. PMID- 17129760 TI - Use of bioimpedance analysis in patients with chronic heart failure? PMID- 17129761 TI - The influence of training with heavy rugby balls on selected spin pass variables in youth rugby union players. AB - The purpose of this study was to investigate the influence of training with heavy rugby balls on selected spin pass variables in youth rugby union players. Pre training, 14 participants performed rapid spin passes (using right and left hands) at a wall-mounted target, from 7-m and 10-m distances. Horizontal (linear) and spin (longitudinal angular) velocities of the passes were recorded using a 25Hz video camera. Participants also performed right- and left-handed spin passes for maximal distance. The participants were then randomly divided into two equal groups, to train twice weekly for eight weeks, using either normal or heavy rugby balls. Each individual performed 50-90 rapid spin passes ranging from 5m to 12m with each hand per session. Following training, participants were reassessed using the same pre-training protocols. Significant changes were found pre-to-post training for both the normal ball, and heavy ball groups, for 7-m right-handed horizontal (linear) velocities (9.80+/-0.45 to 10.27+/-0.82ms(-1), and 9.42+/ 1.04 to 10.19+/-1.03ms(-1), respectively; p=0.029), 10-m left-handed spin (longitudinal angular) velocities (5.13+/-1.60 to 6.08+/-1.30revss(-1), and 4.39+/-1.62 to 5.81+/-0.65revss(-1), respectively; p=0.014), right-handed maximal distance passes (19.0+/-3.6 to 20.6+/-4.0m, and 18.6+/-3.4 to 20.7+/-4.3m, respectively; p=0.001) and left-handed maximal distance passes (15.3+/-3.2 to 18.1+/-2.5m, and 15.0+/-3.0 to 17.6+/-3.7m, respectively; p<0.000). No significant changes could be attributed to training exclusively with the heavy rugby balls. The results of this investigation suggest that intense repetitive passing practice increases a small number of selected spin passing variables in youth rugby union players. PMID- 17129762 TI - Prostaglandins and adenosine in the regulation of sleep and wakefulness. AB - Prostaglandin (PG) D2 and adenosine are potent humoral sleep-inducing factors that accumulate in the brain during prolonged wakefulness. PGD2 is produced in the brain by lipocalin-type PGD synthase, which is localized mainly in the leptomeninges, choroid plexus and oligodendrocytes, and circulates in the cerebrospinal fluid as a sleep hormone. It stimulates DP1 receptors on leptomeningeal cells of the basal forebrain to release adenosine as a paracrine signaling molecule to promote sleep. Adenosine activates adenosine A2A receptor expressing sleep-active neurons in the basal forebrain and the ventrolateral preoptic area. Sleep-promoting neurons in the ventrolateral preoptic area send inhibitory signals to suppress the histaminergic neurons in the tuberomammillary nucleus, which contribute to arousal through histamine H1 receptors. Increased knowledge of the molecular mechanisms by which PGD2 induces sleep through activation of adenosine A2A receptors and inhibition of the histaminergic arousal system will be useful both for a better understanding of sleep/wake regulation and for the development of novel types of sleeping pills or anti-doze drugs. PMID- 17129763 TI - Withdrawal symptoms on display: Bcl-2 members under investigation. AB - The balance between survival and apoptosis of lymphocytes is considered to be regulated by specific signals delivered through cell surface receptors binding either antigen (fragments) or homeostatic cytokines. Expanding lymphocytes must also compete for nutrients. For growth factors and nutrients, recent data indicate how these generic environmental signals couple to members of the apoptosis-regulating Bcl-2 family. The prosurvival molecule Mcl-1 is engaged by lethal BH3-only proteins Puma and Noxa under these circumstances. We propose that Puma and Noxa have specific roles in tipping the balance towards apoptosis after growth factor withdrawal and nutrient shortage, respectively. These complementary mechanisms tune survival in the various niches when lymphocytes compete for resources during selection and expansion. PMID- 17129764 TI - What is immune privilege (not)? AB - The 'immune privilege' of the central nervous system (CNS) is indispensable for damage limitation during inflammation in a sensitive organ with poor regenerative capacity. It is a longstanding notion which, over time, has acquired several misconceptions and a lack of precision in its definition. In this article, we address these issues and re-define CNS immune privilege in the light of recent data. We show how it is far from absolute, and how it varies with age and brain region. Immune privilege in the CNS is often mis-attributed wholly to the blood brain barrier. We discuss the pivotal role of the specialization of the afferent arm of adaptive immunity in the brain, which results in a lack of cell-mediated antigen drainage to the cervical lymph nodes although soluble drainage to these nodes is well described. It is now increasingly recognized how immune privilege is maintained actively as a result of the immunoregulatory characteristics of the CNS-resident cells and their microenvironment. PMID- 17129765 TI - Clinical and electrophysiological features of epilepsy in Italian patients with CLN8 mutations. AB - Neuronal ceroid lipofuscinoses (NCLs) are characterized by epilepsy, visual failure, psychomotor deterioration, and accumulation of autofluorescent lipopigment. CLN8 mutations result in Northern epilepsy and Turkish variant late infantile NCL. We describe the clinical and neurophysiological findings of three patients with CLN8 mutations from Italy. In these patients, the onset of epilepsy occurred between 3 and 6 years of age, with myoclonic, tonic-clonic, and atypical absence seizures. Electroencephalograms revealed focal and/or generalized abnormalities. In all cases, blindness and progressive attenuation of the electroretinogram were observed. Magnetic resonance imaging revealed cerebral and cerebellar atrophy, thinning of the corpus callosum, deep white matter hyperintensity, and hyperintensity of the posterior limb of internal capsules. Skin biopsy revealed lysosomal storage in the cytoplasm of fibroblasts. The clinical picture of our cases resembles that of the Turkish patients and clearly differs from that of Northern epilepsy, which is marked by a prolonged course without myoclonus and visual loss. Definition of the clinical spectrum of this condition will aid in its recognition and have implications for diagnosis and genetic counseling. PMID- 17129767 TI - Base damage and single-strand break repair: mechanisms and functional significance of short- and long-patch repair subpathways. AB - A large variety of DNA lesions induced by environmental agents or arising as an outcome of cellular metabolism are counteracted by a complex network of proteins that belong to the base excision repair/single strand break repair (BER/SSBR) processes. No matter whether the initial lesions are modified DNA bases or single strand breaks with non-conventional termini these processes are completed either by replacement of a single (short-patch, SP) or more (long-patch, LP) nucleotides by different arrays of proteins. Here, the factors that are involved in the selection between SP- and LP-BER/SSBR are reviewed. The biological significance of these alternative subpathways is also presented as inferred from mutant mouse/cell models. PMID- 17129766 TI - Developmental expression of mouse Follistatin-like 1 (Fstl1): Dynamic regulation during organogenesis of the kidney and lung. AB - Follistatin-like 1 (Fstl1) is a distantly related homolog of the Activin and Bone Morphogenetic Protein antagonist Follistatin. Interestingly, this molecule also has homology with the extracellular matrix modifying protein BM 40/SPARC/osteonectin. Previous studies in chick have identified Fstl1 as a regulator of early mesoderm patterning, somitogenesis, myogenesis and neural development. In this study, we determine the developmental expression pattern of Fstl1 in the mouse. We find that Fstl1 is ubiquitously expressed in the early embryo, and that expression becomes regionalized later during development. In the majority of tissues, Fstl1 is strongly expressed in the mesenchymal component and excluded from the epithelium. Notable exceptions include the central nervous system, in which Fstl1 expression is entirely absent with the exception of the choroid plexi and floor plate, the lung, in which Fstl1 expression can be seen in airway epithelia and the kidney, in which collecting ducts and nascent nephron epithelia express the highest levels of Fstl1. PMID- 17129768 TI - Crosstalk of DNA glycosylases with pathways other than base excision repair. AB - While a role for DNA glycosylase activity in base excision repair (BER) is well understood, there is mounting evidence to implicate cooperation of DNA glycosylases with components of repair pathways other than BER. The mechanisms by which DNA glycosylases interact with non-BER pathways are, in many cases, poorly understood. However, accumulating evidence indicates that crosstalk is common and may be as important in signaling repair as the canonical pathways themselves. PMID- 17129769 TI - Positive and negative electrospray LC-MS-MS methods for quantitation of the antiparasitic endectocide drugs, abamectin, doramectin, emamectin, eprinomectin, ivermectin, moxidectin and selamectin in milk. AB - Avermectin endectocides are used for the treatment of cattle against a variety of nematode and arthropod parasites, and consequently may appear in milk after normal or off-label use. The compounds abamectin, doramectin, and ivermectin, contain only C, H and O and may be expected to be detected by LC-MS in negative ion mode. The others contain nitrogen in addition and would be expected to be preferentially ionized in positive mode. The use of positive ion and negative ion methods with electrospray LC-MS-MS were compared. Using negative ion the compounds abamectin, doramectin, ivermectin, emamectin, eprinomectin, and moxidectin gave a curvilinear response and were quantified in raw milk by LC-MS MS with a triethylamine-acetonitrile buffer over the concentration range 1-60 ppb (microg/kg) using selamectin as the internal standard. The limits of detection (LOD) were between 0.19 ppb (doramectin) and 0.38 ppb (emamectin). The compounds gave maximum sensitivity with positive ionisation from a formic acid-ammonium formate-acetonitrile buffer and were detected in milk (LC-MS-MS) also with a curvilinear response over the range 0.5-60 ppb. Although the positive ion signals were larger, with somewhat lower limits of detection (LOD between 0.06 ppb (doramectin) and 0.32 ppb (moxidectin) the negative ion procedure gave a more linear response and more consistent results. Comparison of spiked samples in the range 2-50 ppb showed a high degree of correlation between the two methods. PMID- 17129770 TI - Seeking new antibiotics in nature's backyard. AB - As the pipeline of new antibiotics slows to a trickle, scientists are developing innovative strategies to unearth antibacterial compounds in unexpected places. PMID- 17129771 TI - Re-evaluating cell-cycle regulation by E2Fs. AB - Activation of E2F transcription factors is thought to drive the expression of genes essential for the transition of cells from G1 to S phase and for the initiation of DNA replication. However, this textbook view of E2Fs is increasingly under challenge. Here we discuss an alternative model for how E2Fs may work. PMID- 17129772 TI - Another job for the talented p120-catenin. AB - Given the complexity of signaling pathways in the cell, it is a mystery how these pathways communicate with one other. In this issue of Cell, Wildenberg et al. (2006) reveal that the key effector molecule p120-catenin can mediate crosstalk between the Rac and Rho signaling pathways. PMID- 17129773 TI - ER targeting signals: more than meets the eye? AB - The signal sequences that target newly synthesized proteins to the endoplasmic reticulum are highly variable; however, the functional significance of this diversity has remained obscure. In this issue, Kang et al. (2006) report that variability in signal sequences allows the cell to selectively regulate the translocation of proteins into the endoplasmic reticulum in a substrate-specific manner. PMID- 17129774 TI - A role for p130Cas in mechanotransduction. AB - Focal adhesions are sites of contact between cells and the extracellular matrix. Sawada et al. (2006) now report that the mechanical stretching of cells forces p130Cas, an adaptor protein at focal adhesions, to undergo a conformational change. This change promotes phosphorylation of p130Cas by Src family kinases and the transduction of integrin-mediated signaling. PMID- 17129775 TI - Follow your nose: axon pathfinding in olfactory map formation. AB - Two new studies report how discrete identities of olfactory sensory neurons are converted into a spatial map of axonal connections (Imai et al., 2006; Serizawa et al., 2006). They find that levels of cAMP signals derived from olfactory receptors (ORs) can direct targeting of axons along an axis, and that ORs and neural activity regulate expression of adhesion/guidance molecules in mosaic patterns that can sort axons into discrete locations. PMID- 17129776 TI - Proteins kinases: chromatin-associated enzymes? AB - Protein kinases contribute to the regulation of gene expression by interacting with transcription factors that are recruited to the regulatory regions of genes. Previous studies investigated the role of protein kinases in transcription initiation. Here, we discuss new insights gleaned from recent work showing that kinases can also interact with chromatin throughout the entire transcribed region of target genes (Pokholok et al., 2006; Proft et al., 2006). PMID- 17129777 TI - Pathway to totipotency: lessons from germ cells. AB - Oocytes and sperm are some of the most differentiated cells in our bodies, yet they generate all cell types after fertilization. Accumulating evidence suggests that this extraordinary potential is conferred to germ cells from the time of their formation during embryogenesis. In this Review, we describe common themes emerging from the study of germ cells in vertebrates and invertebrates. Transcriptional repression, chromatin remodeling, and an emphasis on posttranscriptional gene regulation preserve the totipotent genome of germ cells through generations. PMID- 17129778 TI - Tumor morphology and phenotypic evolution driven by selective pressure from the microenvironment. AB - Emergence of invasive behavior in cancer is life-threatening, yet ill-defined due to its multifactorial nature. We present a multiscale mathematical model of cancer invasion, which considers cellular and microenvironmental factors simultaneously and interactively. Unexpectedly, the model simulations predict that harsh tumor microenvironment conditions (e.g., hypoxia, heterogenous extracellular matrix) exert a dramatic selective force on the tumor, which grows as an invasive mass with fingering margins, dominated by a few clones with aggressive traits. In contrast, mild microenvironment conditions (e.g., normoxia, homogeneous matrix) allow clones with similar aggressive traits to coexist with less aggressive phenotypes in a heterogeneous tumor mass with smooth, noninvasive margins. Thus, the genetic make-up of a cancer cell may realize its invasive potential through a clonal evolution process driven by definable microenvironmental selective forces. Our mathematical model provides a theoretical/experimental framework to quantitatively characterize this selective pressure for invasion and test ways to eliminate it. PMID- 17129779 TI - Identification of an intestinal folate transporter and the molecular basis for hereditary folate malabsorption. AB - Folates are essential nutrients that are required for one-carbon biosynthetic and epigenetic processes. While folates are absorbed in the acidic milieu of the upper small intestine, the underlying absorption mechanism has not been defined. We now report the identification of a human proton-coupled, high-affinity folate transporter that recapitulates properties of folate transport and absorption in intestine and in various cell types at low pH. We demonstrate that a loss-of function mutation in this gene is the molecular basis for hereditary folate malabsorption in a family with this disease. This transporter was previously reported to be a lower-affinity, pH-independent heme carrier protein, HCP1. However, the current study establishes that a major function of this gene product is proton-coupled folate transport required for folate homeostasis in man, and we have thus amended the name to PCFT/HCP1. PMID- 17129780 TI - Deletion of DDB1 in mouse brain and lens leads to p53-dependent elimination of proliferating cells. AB - DDB1, a component of the Cul4 ubiquitin ligase complex, promotes protein ubiquitination in diverse cellular functions, including nuclear excision repair, regulation of the cell cycle, and DNA replication. To investigate its physiological significance, we generated mice with null and floxed alleles of the DDB1 gene. Here we report that null mutation of DDB1 caused early embryonic lethality, while conditional inactivation of the gene in brain and lens led to neuronal and lens degeneration, brain hemorrhages, and neonatal death. These defects stemmed from a selective elimination of nearly all proliferating neuronal progenitor cells and lens epithelial cells by apoptosis. The cell death was preceded by aberrant accumulation of cell cycle regulators and increased genomic instability and could be partially rescued by removal of the tumor suppressor protein p53. Our results indicate that DDB1 plays an essential role in maintaining viability and genomic integrity of dividing cells. PMID- 17129781 TI - Structural basis of transcription: role of the trigger loop in substrate specificity and catalysis. AB - New structures of RNA polymerase II (pol II) transcribing complexes reveal a likely key to transcription. The trigger loop swings beneath a correct nucleoside triphosphate (NTP) in the nucleotide addition site, closing off the active center and forming an extensive network of interactions with the NTP base, sugar, phosphates, and additional pol II residues. A histidine side chain in the trigger loop, precisely positioned by these interactions, may literally "trigger" phosphodiester bond formation. Recognition and catalysis are thus coupled, ensuring the fidelity of transcription. PMID- 17129782 TI - Kinetochore microtubule dynamics and attachment stability are regulated by Hec1. AB - Mitotic cells face the challenging tasks of linking kinetochores to growing and shortening microtubules and actively regulating these dynamic attachments to produce accurate chromosome segregation. We report here that Ndc80/Hec1 functions in regulating kinetochore microtubule plus-end dynamics and attachment stability. Microinjection of an antibody to the N terminus of Hec1 suppresses both microtubule detachment and microtubule plus-end polymerization and depolymerization at kinetochores of PtK1 cells. Centromeres become hyperstretched, kinetochore fibers shorten from spindle poles, kinetochore microtubule attachment errors increase, and chromosomes severely mis-segregate. The N terminus of Hec1 is phosphorylated by Aurora B kinase in vitro, and cells expressing N-terminal nonphosphorylatable mutants of Hec1 exhibit an increase in merotelic attachments, hyperstretching of centromeres, and errors in chromosome segregation. These findings reveal a key role for the Hec1 N terminus in controlling dynamic behavior of kinetochore microtubules. PMID- 17129783 TI - The conserved KMN network constitutes the core microtubule-binding site of the kinetochore. AB - The microtubule-binding interface of the kinetochore is of central importance in chromosome segregation. Although kinetochore components that stabilize, translocate on, and affect the polymerization state of microtubules have been identified, none have proven essential for kinetochore-microtubule interactions. Here, we examined the conserved KNL-1/Mis12 complex/Ndc80 complex (KMN) network, which is essential for kinetochore-microtubule interactions in vivo. We identified two distinct microtubule-binding activities within the KMN network: one associated with the Ndc80/Nuf2 subunits of the Ndc80 complex, and a second in KNL-1. Formation of the complete KMN network, which additionally requires the Mis12 complex and the Spc24/Spc25 subunits of the Ndc80 complex, synergistically enhances microtubule-binding activity. Phosphorylation by Aurora B, which corrects improper kinetochore-microtubule connections in vivo, reduces the affinity of the Ndc80 complex for microtubules in vitro. Based on these findings, we propose that the conserved KMN network constitutes the core microtubule binding site of the kinetochore. PMID- 17129784 TI - Substrate-specific translocational attenuation during ER stress defines a pre emptive quality control pathway. AB - Eukaryotic proteins entering the secretory pathway are translocated into the ER by signal sequences that vary widely in primary structure. We now provide a functional rationale for this long-observed sequence diversity by demonstrating that differences among signals facilitate substrate-selective modulation of protein translocation. We find that during acute ER stress, translocation of secretory and membrane proteins is rapidly and transiently attenuated in a signal sequence-selective manner. Their cotranslational rerouting to the cytosol for degradation reduces the burden of misfolded substrates entering the ER and represents a pathway for pre-emptive quality control (pQC). Bypassing the pQC pathway for the prion protein increases its rate of aggregation in the ER lumen during prolonged stress and renders cells less capable of viable recovery. Conversely, pharmacologically augmenting pQC during ER stress proved protective. Thus, protein translocation is a physiologically regulated process that is utilized for pQC as part of the ER stress response. PMID- 17129785 TI - Force sensing by mechanical extension of the Src family kinase substrate p130Cas. AB - How physical force is sensed by cells and transduced into cellular signaling pathways is poorly understood. Previously, we showed that tyrosine phosphorylation of p130Cas (Cas) in a cytoskeletal complex is involved in force dependent activation of the small GTPase Rap1. Here, we mechanically extended bacterially expressed Cas substrate domain protein (CasSD) in vitro and found a remarkable enhancement of phosphorylation by Src family kinases with no apparent change in kinase activity. Using an antibody that recognized extended CasSD in vitro, we observed Cas extension in intact cells in the peripheral regions of spreading cells, where higher traction forces are expected and where phosphorylated Cas was detected, suggesting that the in vitro extension and phosphorylation of CasSD are relevant to physiological force transduction. Thus, we propose that Cas acts as a primary force sensor, transducing force into mechanical extension and thereby priming phosphorylation and activation of downstream signaling. PMID- 17129786 TI - p120-catenin and p190RhoGAP regulate cell-cell adhesion by coordinating antagonism between Rac and Rho. AB - Integration of receptor tyrosine kinase, integrin, and cadherin activities is crucial for normal cell growth, motility, and adhesion. Here, we describe roles for p120-catenin (p120) and p190RhoGAP that coordinate crosstalk between these systems and regulate cadherin function. Surprisingly, PDGFR-induced actin remodeling in NIH3T3 cells is blocked in the absence of p120, and the cells are partially transformed via constitutive activation of Rho. We have traced the mechanism to unexpected codependent roles for p120 and p190RhoGAP in regulating Rac-dependent antagonism of Rho. Receptor-induced Rac activity causes translocation of p190RhoGAP to adherens junctions (AJs), where it couples to the cadherin complex via interaction with p120. AJ formation is dependent on this p120-p190RhoGAP interaction and fails altogether if either of these proteins are compromised. We propose that Rac activation links diverse signaling systems to AJ assembly by controlling transient p190RhoGAP interactions with p120 and localized inhibition of Rho. PMID- 17129787 TI - GATA-3 maintains the differentiation of the luminal cell fate in the mammary gland. AB - The GATA family of transcription factors plays fundamental roles in cell-fate specification. However, it is unclear if these genes are necessary for the maintenance of cellular differentiation after development. We identified GATA-3 as the most highly enriched transcription factor in the mammary epithelium of pubertal mice. GATA-3 was found in the luminal cells of mammary ducts and the body cells of terminal end buds (TEBs). Upon conditional deletion of GATA-3, mice exhibited severe defects in mammary development due to failure in TEB formation during puberty. After acute GATA-3 loss, adult mice exhibited undifferentiated luminal cell expansion with basement-membrane detachment, which led to caspase mediated cell death in the long term. Further, FOXA1 was identified as a downstream target of GATA-3 in the mammary gland. This suggests that GATA-3 actively maintains luminal epithelial differentiation in the adult mammary gland, which raises important implications for the pathogenesis of breast cancer. PMID- 17129788 TI - A neuronal identity code for the odorant receptor-specific and activity-dependent axon sorting. AB - In the mouse, olfactory sensory neurons (OSNs) expressing the same odorant receptor (OR) converge their axons to a specific set of glomeruli in the olfactory bulb. To study how OR-instructed axonal fasciculation is controlled, we searched for genes whose expression profiles are correlated with the expressed ORs. Using the transgenic mouse in which the majority of OSNs express a particular OR, we identified such genes coding for the homophilic adhesive molecules Kirrel2/Kirrel3 and repulsive molecules ephrin-A5/EphA5. In the CNGA2 knockout mouse, where the odor-evoked cation influx is disrupted, Kirrel2 and EphA5 were downregulated, while Kirrel3 and ephrin-A5 were upregulated, indicating that these genes are transcribed in an activity-dependent manner. Mosaic analysis demonstrated that gain of function of these genes generates duplicated glomeruli. We propose that a specific set of adhesive/repulsive molecules, whose expression levels are determined by OR molecules, regulate the axonal fasciculation of OSNs during the process of glomerular map formation. PMID- 17129789 TI - SnapShot: Cellular bodies. PMID- 17129792 TI - Eye-movement study and human performance using telepathology virtual slides: implications for medical education and differences with experience. AB - A core skill in diagnostic pathology is light microscopy. Remarkably little is known about human factors that affect the proficiency of pathologists as light microscopists. The cognitive skills of pathologists have received relatively little attention in comparison with the large literature on human performance studies in radiology. One reason for this lack of formal visual search studies in pathology has been the physical restrictions imposed by the close positioning of a microscope operator's head to the microscope's eyepieces. This blocks access to the operator's eyes and precludes assessment of the microscopist's eye movements. Virtual slide microscopy now removes this barrier and opens the door for studies on human factors and visual search strategies in light microscopy. The aim of this study was to assess eye movements of medical students, pathology residents, and practicing pathologists examining virtual slides on a digital display monitor. Whole histopathology glass slide digital images, so-called virtual slides, of 20 consecutive breast core biopsy cases were used in a retrospective study. These high-quality virtual slides were produced with an array-microscope equipped DMetrix DX-40 ultrarapid virtual slide processor (DMetrix, Tucson, Ariz). Using an eye-tracking device, we demonstrated for the first time that when a virtual slide reader initially looks at a virtual slide his or her eyes are very quickly attracted to regions of interest (ROIs) within the slide and that these ROIs are likely to contain diagnostic information. In a matter of seconds, critical decisions are made on the selection of ROIs for further examination at higher magnification. We recorded: (1) the time virtual slide readers spent fixating on self-selected locations on the video monitor; (2) the characteristics of the ways the eyes jumped between fixation locations; and (3) x and y coordinates for each virtual slide marking the sites the virtual slide readers manually selected for zooming to higher ROI magnifications. We correlated the locations of the visually selected fixation locations and the manually selected ROIs. Viewing profiles were identified for each group. Fully trained pathologists spent significantly less time (mean, 4.471 seconds) scanning virtual slides when compared to pathology residents (mean, 7.148 seconds) or medical students (mean, 11.861 seconds), but had relatively prolonged saccadic eye movements (P < .0001). Saccadic eye movements are defined as eye movements between fixation locations. On the other hand, the pathologists spent significantly more time than trainees dwelling on the 3 locations they subsequently chose for zooming. Unlike either the medical students or the residents, the pathologists frequently choose areas for viewing at higher magnification outside of areas of foveal (central) vision. Eye movement studies of scanning pathways (scan paths) may be useful for developing eye movement profiles for individuals and for understanding the difference in performances between novices and experts. They may also be useful for developing new visual search strategies for rendering diagnoses on telepathology virtual slides. PMID- 17129794 TI - Advanced methods for handling and preparation of stallion semen. AB - Clinical reproduction in the horse more closely parallels human clinical reproduction than in other domestic farm animals. Horse breeders rarely include fertility as a selection criterion when making mating decisions; in most breeds, there is no licensing or approval of stallions. This has led to a significant number of stallions in the breeding pool that possess desirable performance traits but are subfertile for a variety of reasons, some of them genetically transmitted between generations. Therefore, semen characteristics can vary greatly among stallions within the breeding population. A champion stallion is not gelded or culled for poor semen quality or the inability of his spermatozoa to withstand semen preservation techniques. Rather, equine theriogenologists go to great lengths to maximize reproductive performance using any and all means available. Therefore, advanced methods for processing and selecting stallion semen provide the clinician with valuable tools for handling poor-quality semen or for obtaining spermatozoa for assisted reproduction procedures. PMID- 17129795 TI - Collection and freezing of epididymal stallion sperm. AB - The ability to harvest and preserve epididymal sperm from a stallion after simple elective castration, a catastrophic injury, or severe acute illness and subsequent death has been realized, allowing for the preservation of genetics that would have been lost otherwise.Currently, the care taken to collect the testes and epididymides properly, coupled with proper packaging and shipping, could make the greatest contribution to salvaging viable sperm. As advances in assisted reproductive techniques continue, more offspring may be obtained from stored epididymal sperm from valuable stallions. PMID- 17129796 TI - Sperm morphology in stallions: ultrastructure as a functional and diagnostic tool. AB - Conventional light microscopic evaluation of a seminal ejaculate does not fully avail potential indicators of functional impairment in spermatozoal organelles. The technique of critical quantitative evaluation of morphologic features of individual structural components of spermatozoa at a light microscopic level in conjunction with critical qualitative evaluation of spermatozoal organelles at an ultrastructural level, as described in this article, is a valuable clinical tool. Compared with a battery of sperm function assays used in human andrology clinics, this relatively less expensive and simple technique is an efficient functional and diagnostic tool. PMID- 17129797 TI - Advanced insemination techniques in mares. AB - Advanced artificial insemination techniques, such as deep uterine,hysteroscopic, oviductal, and intrafollicular insemination, are described in the context of the different types of spermatozoa that are now available for insemination, including fresh, chilled, frozen,sex-sorted, and epididymal spermatozoa. The implementation of these new technologies answers and poses questions about the interactions of sperm and oocytes in vivo. PMID- 17129798 TI - Breeding-induced endometritis in mares. AB - Endometritis is a common cause of infertility in broodmares. In the past, the condition was believed to be exclusively the result of bacterial contamination of the uterus. Treatment strategies were focused on preventing bacteria from entering the uterus and on treating mares with signs of endometritis with antibiotics. More recent research on uterine defense mechanisms has increased our understanding of the pathophysiology of equine endometritis. Additional causative agents have been identified, and we have learned to separate uterine infections and a physiologic breeding-induced endometritis resulting from uterine exposure to semen.vi PMID- 17129799 TI - Management of postfixation twins in mares. AB - Methods to manage twins after fixation include natural reduction,dietary reduction, transvaginal ultrasound-guided aspiration,surgical removal, craniocervical dislocation, and transabdominal ultrasound-guided injection. Of these, results have been inconsistent with regard to producing a single healthy foal, except for craniocervical dislocation. This new technique enables the twin to be reduced before complete placenta formation has occurred, allowing the remaining fetus to use the entire endometrial surface and grow to its full potential. PMID- 17129800 TI - Hormone profiles and treatments in the late pregnant mare. AB - Evaluation of hormone profiles in late pregnancy is one of the major determinants of fetoplacental compromise in equine clinical practice. Use of hormone therapies is subjective and reflects, to a large extent, our lack of understanding about the endocrine relations between the mare, placenta, and fetus. This article describes the normal endocrine events in late gestation, the abnormal hormone patterns associated with fetoplacental dysfunction, and the hormone interventions that are currently used or could be used to improve pregnancy outcome. PMID- 17129801 TI - Diagnosis of the compromised equine pregnancy. AB - Identification of a compromised pregnancy in the mare requires the exhaustive collection of a database that includes past and recent reproductive and medical histories and a variety of parameters indicating fetal distress and possibly suggesting neonatal compromise.Judicious interpretation of findings and serial recording of data throughout gestation may help in the early detection of abnormal fetomaternal exchange pathways. Some sources of compromise may be identified, and the impact on fetomaternal well-being maybe calculated. Appropriate preventive or corrective measures may then be implemented to minimize the risks of an unfavorable outcome. PMID- 17129802 TI - Diagnosis and treatment of equine placentitis. AB - Equine placentitis is a complex disease that has devastating consequences for horse owners. Placentitis is a significant cause of foal loss annually. Prompt diagnosis and treatment of the disease are paramount for survival of the affected neonate. This article discusses current information on pathogenesis of the disease as well as diagnostic and therapeutic options. PMID- 17129803 TI - Laparoscopic cryptorchidectomy and ovariectomy in horses. AB - Laparoscopic surgery has become commonplace in the field of equine urogenital surgery. As with most surgical procedures, the limiting factors in developing new surgical techniques are limited to the patient size and demeanor, the skills of the surgeon, and the available equipment. Some of the greatest benefits of laparoscopic surgery in the horse include better visualization of the important structures; tension-free amputation of the testes or ovaries,which generally leads to less postoperative pain; and the ability to evaluate the transected stump carefully to make sure there is no hemorrhage. This article is limited to the use of laparoscopy for cryptorchidectomy and ovariectomy. PMID- 17129804 TI - Granulosa cell tumors of the equine ovary. AB - The granulosa cell tumor is the most common ovarian tumor in mares. A clinical diagnosis can be made based on the presence ofa unilaterally enlarged ovary and a small inactive contralateral ovary. Endocrine testing may be beneficial to confirm a diagnosis. Surgical removal of the tumor eliminates the adverse effect on pituitary function and results in resumption of follicular development and ovulation in the opposite ovary over time. PMID- 17129805 TI - Superovulation in mares. AB - Recently, a commercial product has been made available (equine follicle stimulating hormone [eFSH]) for superovulating mares. This has provided the practitioner with a hormonal product that is readily available for enhancing multiple ovulations. Additional benefits of stimulating multiple follicles include an increased number of follicles available for oocyte collection, availability of extra embryos for embryo freezing, enhancement of fertility in subfertile mares, and advancement of the first ovulation of the year. This article provides a short historical background, but most of it centers on the use of eFSH for stimulation of follicular development and ovulation in mares. PMID- 17129806 TI - Vitrification of equine embryos. AB - Vitrification can be used successfully to cryopreserve equine embryos. Embryos for vitrification should be collected from donor mares' uteri when they are 300 mm or less in diameter, however,and at the morula or early blastocyst stage of development. No special equipment is required for vitrification; the straw containing the embryo is exposed to vapor for 1 minute before plunging it into liquid nitrogen. Warming of the straw requires no special equipment,and the embryo can be transferred directly from the straw into a recipient's uterus. Vitrification has been repeatedly successful when the procedure is used with small embryos and provides a method for the rapid cryopreservation of equine embryos. PMID- 17129807 TI - Collection, evaluation, and use of oocytes in equine assisted reproduction. AB - Assisted reproductive techniques have been developed to obtain pregnancies from subfertile mares and stallions and to salvage gametes after death. In recent years, these procedures have been used for clinical cases with repeated success. Although new developments occur, the basis for the success and future development of assisted reproductive techniques is our ability to collect and handle the equine oocyte successfully. This article focuses on important clinical aspects of oocyte collection and evaluation and briefly discusses the clinical use of assisted reproductive procedures in the horse. PMID- 17129808 TI - Equine cloning. AB - Equine cloning is now in use as a clinical technique. It is available commercially, and its efficiency seems to be increasing. The foals produced by cloning may differ in some phenotypic and behavioral traits from the original animal but should produce offspring that reflect those that the original donor animal would have produced. This is especially true in the case of male animals, where the mitochondrial DNA is not passed to the progeny. Results of pregnancies due in 2006 should add significantly to our understanding of the factors affecting production of viable cloned foals and of the similarities and differences among cloned foals and between these foals and the donor animals. PMID- 17129810 TI - Biologics for inflammatory bowel disease: drug approval and monitoring in the United States. AB - Biologics are a primary focus for research and development of new treatments for IBD and other immune disorders. CDER, which is a branch of the FDA, oversees most of the biologics that are being used or tested for IBD, and it regulates nonbiologic medications for IBD. The approval process for most biologics for IBD is similar to the process for nonbiologics. FDA regulation of a new drug or biologic can be divided into three stages: the commercial IND, the NDA, and postmarketing (phase 4) studies. It is unclear if generic versions of biologics can be approved within the current legislation. PMID- 17129811 TI - Focus on mechanisms of inflammation in inflammatory bowel disease sites of inhibition: current and future therapies. AB - Anti-TNF antibodies were the first biologic agents registered to treat patients who have CD and, more recently, patients who have UC. The sequence of events underlying the inflammatory reaction in IBD is extremely complex, however, and involves both the innate and antigen-driven adaptive immune system. Novel therapies are directed at several key players of this cascade. Blockade of T-cell proliferation and activation and inhibition of T-cell cytokines has been most extensively targeted by clinical trials in humans. Inhibition of adhesion molecules and the use of selected growth factors seem to have therapeutic potential. Restoration of regulatory T-cell and dendritic-cell function is still waiting to be explored in clinical trials. Although an increasing number of biologic therapies for IBD are being developed, the discovery of the full spectrum of treatment modalities is only beginning. Often, however, the clinical efficacy of biologic agents is investigated, and for some molecules is established, before mechanisms of action are specifically explored. Eight years after the Food and Drug Administration approved infliximab for the treatment of luminal CD, it is not known how this anti-TNF antibody actually dampens inflammation in IBD. The advent of newer anti-TNF agents is only postponing the answer. PMID- 17129812 TI - General principles and pharmacology of biologics in inflammatory bowel disease. AB - The pharmacology of each biologic agent is important regarding the dose required to achieve benefits, duration of therapeutic effect, and the induction of immunogenicity. Comprehension of the individual pharmacology, pharmacodynamics, and pharmacokinetics, in addition to the impact of concomitant immunomodulation on immunogenicity allows optimization of each biologic agent in the appropriate inductive or maintenance setting of IBD. PMID- 17129813 TI - Infliximab use in luminal Crohn's disease. AB - Infliximab has been available in the United States and Europe for more than 6 years, and its use has revolutionized the care of patients who have CD. It is used effectively for both the induction and maintenance of remission in patients who have CD and is efficacious in patients who have steroid-dependent/refractory CD and those who have fistulizing CD. Clinical trials and practice have shown infliximab to be safe, effective, and generally well tolerated. The ACCENT I and ACCENT II trials defined the best dosing and schedule regimens for its administration. With up to 30% of patients not responding to infliximab therapy, much attention has been devoted to identifying risk factors that could allow optimization of response rates. Parsi and colleagues and Arnott and colleagues demonstrated that nonsmoking and the concurrent use of immunomodulators are predictors of response to infliximab. Research has also focused on identifying biologic and immunologic markers that may correlate with response to infliximab. To date, N0D2/CARD15, anti-Saccharomyces cerevisiae antibody (ASCA), and antineutrophil cytoplasmic antibody (ANCA) have not been shown to be predictive of outcome with infliximab treatment for CD. Gene polymorphisms also are being studies with the hope that knowing the patient's genotype may help predict the course or severity of the disease, including the presence of extraintestinal manifestations, response to treatments, and susceptibility to toxicities. No single variable, however, has been consistently demonstrated to be a predictor of response to infliximab. The formation of ATIs in a small number of patients creates a clinical dilemma. ATIs have been associated with an attenuated response or loss of response to the medication over time and the development of both acute and delayed infusion reactions that occasionally are severe enough to lead to discontinuation of the medication. In such patients physicians are often left to ponder what therapy to try next. Adalimumab, a fully human monoclonal antibody used for treating rheumatologic conditions, has been investigated as an alternate treatment for patients who have CD who, after initially responding to infliximab, experience intolerance or loss of efficacy. Two studies have examined the use of adalimumab in patients who have active CD who had lost response to or developed intolerance to infliximab. In both these studies adalimumab was well tolerated and seemed to be a clinically beneficial option for such patients. Confirmation of these findings with ongoing randomized, double-blind, placebo-controlled trials is needed, however. The limits of conventional treatment for CD can be seen as a positive evolutionary force favoring the development and use of advanced therapies. The acceptance of antimetabolites began with data published a quarter-century ago and became robust in the past 5 to 10 years. Biologic therapy has become the standard of care at a far faster rate. The success seen with infliximab has broadened the acceptance of biologic therapy among professional peers, patients, and pharmaceutical developers. The lessons learned in the years since infliximab's arrival show the importance of long-term data in revealing important toxicities and best practices for maintenance. Tempered by this experience, the short cycle from concept to drug production possible with biologic therapies should bring even more advanced treatments to patients quickly while investigators work to find a cure. PMID- 17129814 TI - Infliximab in fistulizing Crohn's disease. AB - The treatment of fistulizing CD has evolved greatly in the last 15 years, largely caused by improvements in medical therapy. Tables 2 and 3 summarize all published controlled and uncontrolled trials of immunomodulator and biologic therapy for the treatment of Crohn's fistulae. The advent of immunomodulators and anti-TNF alpha agents has transformed the treatment of Crohn's fistulae from almost exclusively surgical to placing a much larger emphasis on medical therapy, either as initial therapy alone, with surgery reserved for refractory cases, or in combination with surgery from the start. For this reason, surgeons and gastroenterologists must work in concert to provide the best care for each patient. Proper fistula management also relies heavily on accurate diagnosis, especially defining the anatomy of the fistula, ascertaining whether abscess formation is present, and determining the location and extent of intestinal inflammation. PMID- 17129815 TI - Infliximab in ulcerative colitis. AB - Infliximab is effective for treatment of moderate-to-severe UC and is recommended for patients who have had an inadequate response to medical therapy or who are intolerant of or do not desire to take the potential risk of using specific agents including immunomodulators (cyclosporine A, azathioprine, or 6 mercaptopurine), corticosteroids, and, potentially, mesalamine. Future trials are needed to assess the efficacy of infliximab with immunomodulators to see if additional benefit is achieved so that the risk-benefit ratio is positive. Based on the favorable efficacy of infliximab for UC therapy, the ground work has been established for evaluating infliximab and addressing some of the many unanswered questions and also for assessing other anti-TNF agents and streamlining the anti TNG antibody to improve efficacy, reduce side effects, and ease administration. PMID- 17129816 TI - Safety of infliximab and other biologic agents in the inflammatory bowel diseases. AB - In many ways, infliximab has drastically altered expectations for medical therapy in IBD, and it is expected that adalimumab and certolizumab pegol with ultimately have a similar role. Patients initiating such therapy should be made cognizant of the potential risks of serious infection including opportunistic ones, such as TB and histoplasmosis; demyelinating disorders; CHF; and lymphoma. Proper selection of candidates for anti-TNF-alpha therapy is critical in maintaining a proper benefit-to-risk ratio. PMID- 17129817 TI - Infusion reactions and their management. AB - Infliximab therapy should be optimized to minimize immunogenicity, to prevent infusion reactions, and to maintain clinical response. Based on best available evidence, strategies include minimizing the formation of ATI by administering infliximab as a multidose induction therapy followed by scheduled maintenance regiment, the use of concurrent immunomodulators, and possibly premedicating with steroids. Infusion reaction are common and they can be managed using specific protocols outlined in this article. PMID- 17129818 TI - Economics of the use of biologics in the treatment of inflammatory bowel disease. AB - The introduction of the biologic therapies into the therapeutic regiment for IBD, coupled with the vast changes seen in health care delivery within the past decade, have turned previous economic assumptions and models on their head, and opened up a new opportunity to redefine what is truly the cost of a disease, and of its' interventions. Pundits and pencil-pushers are quick to point to the high drug cost as a threat to the economic stability of the health care system, without first doing a carefully planned, balanced analysis of the overall impact that such a therapy will make on the entire cost-structure of the system. This is also true outside of the United States, where those who must budget and pay for the therapies do not pay for the use of health care services, and simply enter the drug costs on an accountant's debit sheet. Such analyses are complicated, because they must calculate the impact that such therapies have on the direct costs of health care and the indirect costs for both the patient and their families or health care support system. Realizing what impact these therapies might have on altering the natural history of these previously relentless chronic debilitating conditions, and redefining how normal quality of life can actually get on these therapies, should be the focus of future studies as clinicians try to calculate truly whether these agents are cost savings for society overall. PMID- 17129819 TI - Treatment of immune-mediated extraintestinal manifestations of inflammatory bowel disease with infliximab. AB - The introduction of infliximab into clinical practice is one of the most significant advances in the care of patients who have IBD. Infliximab has become an important part of the medical armamentarium to treat extraintestinal manifestations that often are refractory to other medications and are a significant cause of morbidity in these patients. Two other TNF inhibitors recently have demonstrated efficacy in CD: certolizumab pegol and adalimumab. The Food and Drug Administration has approved adalimumab for use in RA. One predicts that these agents also may have activity in the extraintestinal manifestation for IBD. To determine whether future biologics are effective in the EIM of IBD, one may need to look no further than the vast clinical trial experience in primary chronic inflammatory diseases of the joints and skin: RA and psoriasis. For example, the Food and DRug Administration recently has approved an anti-B-cell therapy, rituximab, and a T-cell costimulation modulator, abatacept, for use in RA. It certainly will be of interest to determine whether these biologic agents demonstrate efficacy in the intestinal and EIM of IBD. PMID- 17129820 TI - Hepatitis B: the pathway to recovery through treatment. PMID- 17129821 TI - Saving big pharma from drowning in the data pool. PMID- 17129822 TI - Similarity-based virtual screening using 2D fingerprints. AB - This paper summarizes recent work at the University of Sheffield on virtual screening methods that use 2D fingerprint measures of structural similarity. A detailed comparison of a large number of similarity coefficients demonstrates that the well-known Tanimoto coefficient remains the method of choice for the computation of fingerprint-based similarity, despite possessing some inherent biases related to the sizes of the molecules that are being sought. Group fusion involves combining the results of similarity searches based on multiple reference structures and a single similarity measure. We demonstrate the effectiveness of this approach to screening, and also describe an approximate form of group fusion, turbo similarity searching, that can be used when just a single reference structure is available. PMID- 17129823 TI - Novel fluorescent proteins for high-content screening. AB - The development of fast microscopic imaging devices has enabled the application of automated fluorescence microscopy to pharmaceutical high-throughput drug discovery assays, referred to as high-content screening (HCS). Initially, green fluorescent protein and its derivatives from Aequorea Victoria, and later anthozoan fluorescent proteins (FPs) have become potent tools as live-cell markers in HCS assays. We illustrate the broad applicability of classic and novel FPs to drug-discovery assays, giving example applications of the use of FPs in multiplexed imaging as fluorescent timers, photosensitizers and pulse-chase labels, and for robotically integrated compound testing. PMID- 17129824 TI - Optical biosensors: where next and how soon? AB - From a direct comparison of the technical benefits of labelled reporter assays with the benefits of label-free assays, label-free appears to have significant advantages. Faster assay development times; accurate, high information content data; and less interference from labels. However, optical label-free platforms have not yet made a major impact in the drug discovery technology markets; are often viewed as having poor throughput, limited application; and are difficult to learn and use effectively. PMID- 17129825 TI - Non-optical screening platforms: the next wave in label-free screening? AB - The use of optical biosensors for compound screening was first demonstrated in the mid-1990s, but there has been limited uptake in the market owing to issues of limited throughput and a lack of applications for key receptor classes. Recently, several start-up and established tools companies have exploited non-optical detection modalities that seek to address the shortcomings of more established optical approaches. Platforms based on acoustic resonance, electrical impedance, microcantilevers, nanowires and differential calorimetry are beginning to appear with commercially available products targeted at post-high-throughput screening hit confirmation and mode-of-action studies. This article highlights key advances in commercial label-free analysis platforms, which complement more traditional optical system and which also allow novel assay formats for the analysis of previously intractable targets. PMID- 17129826 TI - The role of the informatics framework in early lead discovery. AB - Recent developments in screening technologies and data analysis have been driven by promises that the numbers of new lead compounds will increase. Although many of these promises have become reality, the success of this strategy also depends on the information framework that ties the individual components together. In particular, high-content technologies represent a new force in challenging established informatics frameworks; largely because of their data volume, variety of assay parameters and increased scientific complexity. A successful informatics framework design can be regarded as crucial for new technologies, both in terms of scientific content and information, and process integration across large corporate networks. PMID- 17129827 TI - Systems biology, metabolic modelling and metabolomics in drug discovery and development. AB - Unlike signalling pathways, metabolic networks are subject to strict stoichiometric constraints. Metabolomics amplifies changes in the proteome, and represents more closely the phenotype of an organism. Recent advances enable the production (and computer-readable encoding as SBML) of metabolic network models reconstructed from genome sequences, as well as experimental measurements of much of the metabolome. There is increasing convergence between the number of human metabolites estimated via genomics ( approximately 3000) and the number measured experimentally. It is thus both timely, and now possible, to bring these two approaches together as an integrated (if distributed) whole to help understand the genesis of metabolic biomarkers, the progress of disease, and the modes of action, efficacy, off-target effects and toxicity of pharmaceutical drugs. PMID- 17129828 TI - Imaging Alzheimer's disease pathology: one target, many ligands. AB - Over the past five years there has been a surge of interest in using positron emission tomography (PET) to determine the in vivo density of the senile plaque, a key pathological feature of Alzheimer's disease. The development of the tracers [(11)C]-PIB, [(11)C]-SB13 and [(18)F]-FDDNP has coincided with drug strategies aimed at altering the brain metabolism of amyloid-beta peptides. The evolution of these novel ligands serves not only as an excellent example of how rapidly imaging technologies can progress but also as a reminder that the fundamental biological knowledge, which is necessary to fully interpret the PET data, can be left trailing behind. PMID- 17129829 TI - Therapeutic potential of interfering with apelin signalling. AB - The apelin receptor is a G protein-coupled receptor activated by several apelin fragments. Its tissue distribution suggests that apelin signalling is involved in a broad range of physiological functions. Endothelial cells, which express high levels of apelin receptors, respond to apelin through the phosphorylation of key intracellular effectors associated with cell proliferation and migration. In addition, apelin is a mitogen for endothelial cells and exhibits angiogenic properties in matrigel experiments. This review focuses on the therapeutic potential of apelin signalling, which is associated with pathologies that result from decreased vascularisation (ischemias) or neovascularisation (retinopathies and solid tumors). PMID- 17129830 TI - Knowledge-based chemoinformatic approaches to drug discovery. AB - The modern drug discovery process is steadily becoming more information driven. Structural, physicochemical and ADME-Tox property profiles of reference (successful) ligands, along with structural information of their target proteins, have been extremely useful for early-stage drug discovery. Recently, databases of known biologically active ligands (knowledge bases) have become more focused toward different protein-target classes. The number of new chemoinformatics tools used to analyze structures and properties of successful molecules has also increased enormously. Scientists in this area are exploring new physicochemical properties and appropriate drug sets to understand druglike properties. In this review, the various uses of the ligand knowledge bases in the drug discovery process have been critically reviewed. PMID- 17129831 TI - Food for thought. AB - Do certain kinds of food contain pharmacologically active substances in concentrations that are high enough to have druglike effects when consumed? Are biologically active compounds in food indicative of therapeutic value? Is traditional drug development suitable for testing the merits of food? Is it ethical to test food as a drug on patients? Will dietary disease management remain a pipedream? Is it a fact or fantasy that the Mediterranean diet is beneficial to health? Is a vegetarian diet an elimination therapy, or one of supplementation? What can be learned from animals? Are humans losing the capability of listening to their bodies? In this review, we will address these questions--providing food for thought. PMID- 17129833 TI - Vascular catheter-associated infections: a microbiological and therapeutic update. AB - The increasing incidence of central venous catheter (CVC)-related infections can be ascribed to the wider indications to central venous catheterization, to the higher attention to this issue paid by clinicians and microbiologists, and to the patient population referred to hospitals, increasingly characterized by different degrees of immunosuppression and often in critical clinical conditions. This phenomenon implies a higher health care burden and higher related costs, as well as a significant attributable mortality, that varies however according to the pathogen involved. The microorganisms most frequently involved in CVC-related infections are coagulase-negative staphylococci, Staphylococcus aureus, aerobic Gram-negative bacilli, and Candida albicans. In the management of suspected or proven central venous catheter-related infections, several issues need to be addressed: the need to remove the device or the possibility of salvage, the immediate start of calculated antibiotic therapy or the possibility of waiting for results of microbiological diagnostics and proceeding to etiologically-guided therapy. The preferred conservative method is the "Antibiotic-Lock technique" (ALT), based on the endoluminal application of antibacterials at extremely high concentrations in situ for a period of time long enough to ensure bactericidal activity. On the other hand, immediate catheter removal and initiation of appropriate calculated therapy immediately after an adequate diagnostic work-up are strongly recommended in a clinical setting of severe sepsis or septic shock. PMID- 17129834 TI - Alterations in the gyrA and parC genes in Salmonella spp. following in vitro exposure to fluoroquinolones. AB - We characterize alterations in the QRDR fragment of gyrA and parC in Salmonella spp. following repeated exposure to different quinolones in two in vitro models. Mutations in QRDR of gyrA were found in only 25% of the mutants. The most frequent mutations were Asp87(R)Asn and Asp87(R)Tyr. Strains with the former mutation had a higher ciprofloxacin MIC than those with the latter. We did not find mutations in parC. Our data confirm that mutation in the QRDR fragment of gyrA is the mechanism that produces the greatest decrease in fluoroquinolone susceptibility. There is a greater reduction in the ciprofloxacin susceptibility of strains that were originally nalidixic acid-resistant than in that of strains that were originally susceptible, and this confirms that there is a greater likelihood of therapeutic failures with such strains. PMID- 17129835 TI - Antimicrobial resistance of Escherichia coli strains causing community-acquired urinary tract infections among insured and uninsured populations in a large urban center. AB - We assessed the susceptibility of Escherichia coli strains causing communityacquired urinary tract infection (UTI) in a large urban center in Brazil, comparing two different populations (patients with health insurance vs. uninsured). 581 nonduplicate strains of E. coli were isolated. The prevalence of antimicrobial resistance was greater than 20% for ampicillin (51%), trimethoprim sulfamethoxazole (43%), tetracycline (41%) and chloramphenicol (22%). Overall, 12% of the E. coli isolates were resistant to ciprofloxacin. Resistance prevalences to most antimicrobials were similar in the two study populations. Our data provide much needed information on the prevalence of antimicrobial resistance among E. coli causing communityacquired UTI in Brazil. Antimicrobial resistance among strains of E. coli causing community-acquired UTIs was relatively high, particularly resistance to ciprofloxacin. PMID- 17129836 TI - Antimicrobial use and resistance among Gram-negative bacilli in an Italian intensive care unit (ICU). AB - Gram-negative bacilli antimicrobial resistance remains a significant problem for patients in the intensive care unit (ICU). We performed a retrospective analysis of microbiological data and antibiotic consumption over a 4-year period (2000 2003) in an Italian ICU. Pseudomonas aeruginosa and Klebsiella pneumoniae represented approximately 40% of all isolates. The most significant trend in antimicrobial use was an increase in use of 3(rd )generation cephalosporins, imipenem, and ciprofloxacin. A significant trend toward an increase in resistance rates to piperacillin, 3( rd )generation cephalosporins and ciprofloxacin was observed for K. pneumoniae and a positive correlation between resistance and drug usage was evident for K. pneumoniae and piperacillin, cefotaxime, ceftazidime, cefepime, and ciprofloxacin, but not for piperacillin/tazobactam. No statistically significant correlations were evidenced for P. aeruginosa. Trends in resistances were studied also for Serratia spp and Proteus spp. Isolation rates of extended-spectrum beta-lactamase (ESBL)-producing strains in pathogens studied were high, especially for K. pneumoniae (72%, 160/222) and Proteus spp (41%, 18/43). In conclusion, the study showed high resistance among Gram-negative organisms isolated in the ICU and significant ESBL production. A significant correlation between antibiotic consumption and increasing resistance was evident for K. pneumoniae. PMID- 17129837 TI - Combined therapy of teicoplanin and caffeic acid phenethyl ester (CAPE) in the treatment of experimental mediastinitis in the rat. AB - Post-sternotomy mediastinitis affects 1-3% of patients undergoing cardiac surgery and is lethal in 10-47% of these patients. We investigated the effect of an antioxidant/anti-inflammatory agent, caffeic acid phenethyl ester (CAPE), in the attenuation of inflammatory response induced by methicillin-resistant Staphylococcus aureus (MRSA) infection in a rat experimental mediastinitis model. Rats, divided into six equal groups, received MRSA precolonized stainless steel wire pieces implanted into their mediastinal spaces. Control group and CAPE control group received saline and CAPE 10 micromol/kg.day(-1 )respectively, where Group A received a single dose of teicoplanin 24 mg/kg i.m. for the first day and then 12 mg/kg.day(-1) . Group B received teicoplanin as in Group A plus CAPE 10 micromol/kg. day(-1 )intra-peritoneally. Group C received teicoplanin 60 mg/kg i.m. for the first day and then 30 mg/kg.day(-1 )and Group D received teicoplanin as in Group C plus CAPE 10 micromol/kg.day(-1) . By the end of 14 days rats were sacrificed and serum malondialdehyde (MDA), myeloperoxidase (MPO), nitric oxide (NO), urea and creatinine levels were evaluated. Mediastinal organ tissues were collected for histopathological analysis. Infection rates in all the drug-treated groups were lower than the control groups ( P=0.002) but statistical significance was attained only between the groups A and D ( P=0.018). In connective tissues and the peribronchial area polymorphonuclear leukocytic (PNL) infiltration in the treatment groups, although becoming very close, did not reach statistical significance (P =0.053, P=0.075, respectively). PNL infiltration especially in the peribronchial tissues of the Group B animals was found to be significantly less than the Control and CAPE Control groups with P values of 0.013 and 0.010, respectively. MDA and MPO levels were significantly lower in the treatment groups ( P<0.001 and P<0.001 respectively). Levels of the degradation products of NO were lower in treatment groups compared to two control groups (P=0.003, P= 0.005). NO levels in Group D were lowest among all treatment groups ( P=0.001). It has been demonstrated that although bacterial colonization can be controlled in mediastinitis, the inflammatory response persists. The combination of an antioxidant / anti-inflammatory agent, CAPE, added to standard antibiotic therapy might be effective in the treatment of post-sternotomy mediastinitis due to MRSA. PMID- 17129838 TI - Antibiotic prophylaxis in hernia repair and breast surgery: a prospective randomized study comparing piperacillin/tazobactam versus placebo. AB - Although antibiotic prophylaxis is not explicitly indicated for hernia repair and breast surgery, its use for these clean procedures is widely adopted, albeit to a different extent in different countries, often on the personal decision of the individual surgeon. The present study was carried out to compare the efficacy of a single pre-operative dose of piperacillin-tazobactam with placebo in preventing surgical wound infections and to determine the main risk factors associated with infections following two main elective surgical clean procedures such as hernia repair and breast surgery.A total of 501 patients undergoing elective inguinal/femoral hernia repair or breast surgery were enrolled in this prospective randomized clinical study. Patients were randomly assigned to receive preoperative antibiotic prophylaxis or placebo. One dose of piperacillin tazobactam 2.250 g or placebo was administered i.v. 30 minutes prior to the surgical procedure. Using statistical univariate analysis, the following variables were correlated with a higher infection risk: age >40 years, concomitant disease, WBC <3500, surgical wound size >9cm, use of drainages, non prophylaxis. Using multivariate analysis, no antibiotic pre-operative prophylaxis, concurrent chronic diseases, especially diabetes (risk 15 times higher), and length of intervention >45 min (risk 6 times higher) were independent predictors of infection. Finally, patients with postoperative infections had a significantly longer hospitalisation. One pre-operative dose of piperacillin-tazobactam 2.250 g is more effective than placebo in preventing postoperative infections in breast surgery and hernia repair. PMID- 17129839 TI - The influence of local instillation of fusidic acid on the development of microbial complications after lung resection. AB - The efficacy of local instillation of fusidic acid in the prevention of post surgical microbial complications during various types of lung resection was studied. Four hundred ninety two consecutive patients who underwent 504 thoracotomies for non-small cell lung carcinoma during April 1998-May 2004 were reviewed. The 290 patients of the first period who underwent 298 thoracotomies received a chemoprophylactic regimen of intravenous cefuroxime while the 202 patients of the second period who underwent 206 thoracotomies were additionally treated with fusidic acid, irrigated with local instillation into the pleural space, for the prevention of postoperative septic complications. Patients were followed postoperatively for development of septic complications (empyema and bronchopleural fistula) as well as of pneumonia and wound infection. Seventeen patients (5.7%) of the first period developed empyema and 13 fistula (4.4%), whereas only 2 patients (1.0%) of the second period developed empyema and fistula (OR = 5.876; 95% CI, 1.343- 25.716; P = 0.008 and OR = 4.193; 95% CI, 1.003 20.130; P = 0.034, respectively). Cases of pneumonia decreased, but not significantly, from 21 (7.0%) during the first period to 9 (4.4%) during the second period (OR = 1.613; 95% CI, 0.724-3.593; P = 0.257) while cases of wound infection decreased significantly from 19 (6.4%) to 2 (1.0%) (OR = 6.567; 95% CI, 1.513-28.510; P = 0.003). During the first period 23 pathogens were found from cases of empyema and 73 pathogens from cases of pneumonia and wound infection, whereas during the second period 3 and 18 pathogens were respectively found (OR = 5.3; 95% CI, 1.570-17.888; P = 0.003, and OR = 2.804; 95% CI, 1.628-4.838; P <0.001, respectively). These results indicate that local instillation of fusidic acid in the pleural space prior to lung resection seems effective in reducing the rate of septic complications as well as of wound infections. PMID- 17129840 TI - Antibiotic prophylaxis in surgery: an observational prospective study conducted in a large teaching hospital in Naples. AB - This was a 9-month observational prospective study conducted in two steps to evaluate surgical prophylaxis procedures used by surgeons in several departments of the Second University of Naples (SUN). In step 1 (4 months), we collected and analyzed data on surgical interventions and antibiotic prophylaxis. Surgeons were informed of the analysis outcome and were given an antibiotic prophylaxis protocol based on international guidelines. In step 2 (5 months), we collected data on surgical interventions and antibiotic prophylaxis, and compared them with step 1 data. The analysis of 354 forms (step 1) showed that third-generation cephalosporins were the preferred prophylactic antibiotics. The analysis of 369 forms (step 2) showed that ceftriaxone and ampicillin were the most frequently used antibiotics. Surgeons did not comply with guidelines for antibiotic prophylaxis as regards type of antibiotic and treatment duration but implementation of antibiotic prophylaxis protocols resulted in more appropriate and better timing of antibiotic prophylaxis. PMID- 17129841 TI - Antibiotic resistance of uropathogens isolated from Al-Shifa hospital in Gaza Strip in 2002. AB - A survey on clinical isolates from urine samples was conducted and the data obtained were analyzed to determine the most common bacterial causes of urinary tract infection in the Gaza Strip and to evaluate the sensitivity patterns to selected antimicrobials that are commonly used to treat such pathogens. Urine samples were taken for culture from inpatients and outpatients at Al-shifa hospital in Gaza. Urinary tract pathogens were identified and their susceptibility to antimicrobials was determined. Of the 4778 processed urine samples, only 1637 were recorded as positive (34%) and only 1283 were tested for antimicrobial susceptibility. Escherichia coli was the most frequent uropathogen (42%), followed by Klebsiella spp. (12.2%), Pseudomonas spp. < (10.2%), other Gram-negative bacilli (8%), b hemolytic streptococci (7.6%), Enterococcus spp. (6.9%), other Staphylococcus spp. (including S . saprophyticus) (6.5%), Proteus (4.5%), S. aureus (2.1%). Resistance to antimicrobials was extremely alarming. E. coli resistance to amoxicillin reached 97.9%, to piperacillin 78.3%, to doxycycline 90%, to sulfamethoxazole/trimethoprim 63.9% and to cefaclor 42.2%. PMID- 17129842 TI - Very high resistance to amoxicillin in Streptococcus pneumoniae: an epidemiological fact or a technical issue? AB - To explore reproducibility of high amoxicillin minimum inhibitory concentrations (MIC(AMX) ), isolates received during 2002 and 2003 in the National Reference Laboratory of Streptococcus pneumoniae with an amoxicillin MIC of 16 microg/ml (43 strains) and 8 g/ml (12 strains) when singly determined on a routine basis in this center by agar dilution, were retested 10 times by agar dilution and microdilution following NCCLS guidelines, not only using double dilutions but also dilution steps of 2 microg/ml (i.e, 2, 4, 6, 8, 10, 12, 14 and 16 microg/ml). A significant (p<0.05) shift to a higher MIC(AMX )was obtained with microdilution vs. agar dilution. Routine MIC(AMX )of 16 microg/ml were confirmed in 0 strains by agar dilution and in 6 by microdilution, when retested. These 6 strains presented a modal MIC(AMX )value of 10 microg/ml (5 cases) and of 14 micro g/ml (1 case) when using 2 microg/ml microdilution steps. There is low reproducibility of the highest MIC(AMX )values. PMID- 17129843 TI - A strain of Salmonella enterica serovar Virchow isolated in Turkey and carrying a CTX-M-3 extended-spectrum beta-lactamase. AB - A multiply resistant strain of Salmonella enterica subsp. enterica serovar Virchow was isolated in November 2002 from a catheterized patient admitted to the SSK Training Hospital in Ankara, Turkey. This isolate showed an antimicrobial susceptibility pattern compatible with the presence of a CTX-M-type ESBL, namely resistance to cefotaxime, aztreonam and cefepime, and intermediate susceptibility to ceftazidime. On checking for the presence of the bla(TEM), bla(SHV), and bla(CTX-M )resistance genes by PCR, negative results were obtained with the primers specific for SHV and TEM genes, while positive results were obtained with those specific for CTX-M-type genes. After sequencing, the beta-lactamase was identified as CTX-M-3. This is the first report of this enzyme in Salmonella Virchow and represents a further disquieting threat to the therapy of infections caused by Salmonella isolates. PMID- 17129844 TI - Efficacy of adjuvant chemotherapy using oral fluorinated pyrimidines for curatively resected gastric cancer: a meta-analysis of centrally randomized controlled clinical trials in Japan. AB - Adjuvant chemotherapy for gastric cancer has been extensively explored in Japan since the 1950s, and a combination of oral fluorinated pyrimidines (o-FP) and mitomycin C (MMC) has been mainly utilized for adjuvant chemotherapy. However, there is no sufficient evidence for the efficacy of adjuvant therapy. Therefore, we assessed the efficacy of o-FPs over surgery alone (control) by means of a meta analysis of Japanese centrally randomized controlled clinical trials conducted between 1980 and 2005. For inclusion in this study, studies had to compare adjuvant chemotherapy for curatively resected gastric cancer with surgery alone, mainly targeting o-FP, and central randomization designed to comply with contemporary standards for clinical trials in Japan. For the 4 trials that met the eligibility criteria, the estimated hazard ratio was 0.73 (95%CI=0.60-0.89). Our findings show that in Japan adjuvant chemotherapy using o-FP for long-term maintenance therapy appears to be effective for gastric cancer patients after curative resection. PMID- 17129845 TI - Factors influencing the use of antitumoral chemotherapy in the South East of England. AB - Influences on the use of chemotherapy for the treatment of cancer within the South East region of England for patients diagnosed with colorectal, lung, breast and prostate cancer were investigated. The variables investigated as possibly influencing the selection of chemotherapy were the sex of the patients, their age, the year of diagnosis, the cancer site, the cancer stage, the index of multiple deprivation (IMD) and the cancer network of residence. Logistic regression used to adjust the proportion receiving chemotherapy in relation to other variables considered showed significant differences in the proportion of patients receiving chemotherapy between different cancer sites and different networks. There was also a highly significant trend seen in use of chemotherapy over time; the adjusted proportion of patients receiving chemotherapy increasing from 10.6% in 1993 to 24.3% in 2002. Age, stage and cancer site seemed to have the most influence on the use of chemotherapy. PMID- 17129846 TI - A case of Guillain-Barre syndrome in a patient with non small cell lung cancer treated with chemotherapy. AB - Guillain-Barre Syndrome (GBS) is a demyelinating polyneuropathy of probable autoimmune pathogenesis characterized by rapidly progressive symmetric paralysis. In the literature some cases of GBS associated with anticancer chemotherapy are reported. We present a case of a 55-year old woman who complained of progressive motor deficit in four limbs, areflexia in lower limbs and facial nerve paralysis one week after beginning cisplatin-gemcitabine chemotherapy for metastatic lung cancer. The cerebrospinal fluid analysis showed a strong positive Pandy reaction with 435 mg/dl total protein. The electromyography and the electroneuronography established the diagnosis of inflammatory demyelinating polyneuropathy. Specific therapy with intravenous immunoglobulin 25 g/day in 5 administrations for 5 days was started with complete benefit. PMID- 17129847 TI - Antimicrobial resistance--fluoroquinolones and the tiger's tail. PMID- 17129848 TI - Listeria monocytogenes meningitis and multiple brain abscesses in an immunocompetent host. Favorable response to combination linezolid-meropenem treatment. PMID- 17129849 TI - Nosocomial strain of Serratia marcescens producing a new type of extended spectrum beta-lactamase transfers multidrug resistance. PMID- 17129850 TI - Identification of a new AT-rich-element binding factor PsATF1 and its combined effect with PsGBF on the activation of PsCHS1 promoter. AB - Chaltone synthase (CHS) is a key speed-limiting enzyme in the phenylpropanoid pathway which plays an important role in plant defense response against pathogens. In the PsCHS1 promoter, there is an AT-rich element (ATRE) which is required for the maximal elicitor-mediated activation. However, the transcription activator of the ATRE and its regulation mechanism in pea keep unclear. In this paper, a new ATRE-binding factor was isolated from an elicitor-induced pea cDNA expression library and was designated as PsATF1. Electrophoretic mobility shift assay (EMSA) indicated the ATRE-specific binding activity of PsATF1. Beta galactosidase assays in yeast cells suggested that PsATF1 possessed transcription activating activity because PsATF1 activated the expression of the reporter gene even without the GAL4 activation domain (AD). The current study also examined the co-activation effects of PsATF1 with another transcription factor PsGBF on ATRE or PsCHS1 promoter through a transient expression system. The present work reports that PsATF1 acts as a complete transcription activator and first indicates that there are combined effects of PsATF1 with PsGBF on the activation of PsCHS1 promoter. These results provide theoretical basis to the plant defense gene expression mechanism regulated by multiple activators. PMID- 17129851 TI - Ethnicity and risk factors in needle sharing among intravenous drug users in Sichuan Province, China. AB - Combining survey and ethnographic data, this research examined differences in the risk factors associated with needle sharing amongst intravenous drug users (IDUs) in the Sichuan Province of China. A comparison was made between the province's majority Han population and its Yi minority. We developed a theoretical framework consisting of risk factors at the individual level (including risk factors such as lack of AIDS knowledge, low self-efficacy, and economic pressure), interpersonal level (having an IDU primary partner and lack of family support), and community level (social discrimination). The findings suggested that the Yi minority group was more socially disadvantaged and had a higher risk of contracting HIV than the Han group. Furthermore, the factors that put them at risk were different to those which affected the Han group. OLS regression results showed that, for Han IDUs, needle sharing was positively associated with having an IDU primary partner and with economic pressure. On the other hand, for the minority group, needle sharing was significantly associated with being male, AIDS knowledge, the lack of family support, and social discrimination. These findings highlight the need for HIV prevention work to target marginalized populations in China, such as ethnic minorities, and to tailor appropriate prevention strategies to meet the specific needs of different groups. PMID- 17129852 TI - HIV and ethnicity in Canada: is the HIV risk-taking behaviour of young foreign born MSM similar to Canadian born MSM? AB - There is a dearth of information on the HIV risk-taking behaviour of foreign-born men who have sex with men (MSM) in Canada. This study focused on identifying sexual risk behaviour among MSM who immigrated to Canada and compared them to MSM who were born in Canada. Baseline data from the Omega Cohort in Montreal and the Vanguard Project in Vancouver were combined to form four ethnicity/race analytical categories (n = 1,148): White born in Canada (WBIC), White born outside of Canada, non-White born in Canada (NBIC) and non-White born outside of Canada (NBOC). Psychological, demographic and sexual behaviour characteristics of the groups were similar except: NBOC were more likely to be unemployed, less likely to be tattooed, had fewer bisexual experiences and less likely worried of insufficient funds. WBOC were more likely to report unprotected sex with seropositives and more likely to have had unprotected sex while travelling. NBIC were more likely to have ever sold sex and to have had body piercing. WBOC are at high risk of acquiring as well as transmitting HIV. It is important to consider place of birth in addition to ethnicity when developing programmes to prevent the transmission of HIV. PMID- 17129853 TI - Advantages and challenges of using census and multiplier methods to estimate the number of female sex workers in a Chinese city. AB - Using census and multiplier methods to estimate the size of the population of female sex workers (FSWs) in a small city in western China, this study compared the advantages and challenges of the two methods. It was estimated that there were about 1,500 FSWs within the urban area using the census method, which was significantly lower than that estimated by the multiplier method (2,500). Each method has advantages and limitations, and could be applied to different situations. The census method is less time and resource consuming in smaller regions and has a tendency to underestimate, and therefore, the result can be viewed as a low limit. It is useful in a local setting, for example, when estimations are needed for planning HIV/AIDS prevention programmes in a single city. Using existing information or resources, multiplier method could be used to produce estimates for a large geographic area or at a national level. PMID- 17129854 TI - Sexual assault, sexual risks and gender attitudes in a community sample of South African men. AB - Sexual assault against women and HIV infection are both prevalent and related social problems in South Africa. The current study examined hostile attitudes toward women, acceptance of violence against women and masculine ideological beliefs in relation to sexual assault history among men in a Cape Town township in South African. Men (n=435) completed anonymous surveys of sexual assault history, HIV risk history and gender-based attitudes. More than one in five men in this community sample reported that they had either threatened to use force or used force to gain sexual access to a woman in their lifetime. Men with a history of sexual assault were at significantly higher risk for HIV transmission than their non-sexually assaultive counterparts. Men with a history of sexual assault were also more likely to endorse hostile attitudes toward women and were more likely to accept violence against women, although these attitudes and beliefs were prevalent and pervasive across men with and without histories of sexual assault. These findings extend previous research to show that men who have a history of sexual assault also exhibit elevated risks for HIV infection and transmission. Interventions are needed to address hostile attitudes toward women, sexual assault and sexual risks for HIV among South African men. PMID- 17129855 TI - Stigma and social barriers to medication adherence with urban youth living with HIV. AB - Youth adherence to highly active anti-retroviral therapy (HAART) is poor, and little research exists that identifies the reasons youth have difficulty adhering to medications. Given that complete adherence is necessary for favourable health outcomes, it is essential to examine the obstacles youth face in adhering to HAART. The present investigation sought to identify these barriers and to systematically examine the experiences and attitudes youth have towards medications. Twenty-five adolescents and young adults presenting to a public primary care facility for treatment of HIV infection were asked to participate in focus groups which explored their attitudes and experiences around medication adherence. Participants provided richly detailed descriptions of the challenges of managing HIV stigma and their efforts to hide their status from friends, family, doctors, and even themselves. Fifty percent of respondents indicated that they skipped doses because they feared family or friends would discover their status. These results suggest that HIV stigma impacts treatment for youth on several levels, from the accuracy of communication with medical providers to medication adherence, subsequent health outcomes, and the emergence of treatment resistant strains. PMID- 17129856 TI - Establishing a workplace antiretroviral therapy programme in South Africa. AB - Ways to expand access to antiretroviral treatment (ART) in low income settings are being sought. We describe an HIV care programme including ART in an industrial setting in South Africa. The programme uses guidelines derived from local and international best practice. The training component aims to build capacity among health care staff. Nurses and doctors are supported by experienced HIV clinicians through telephone consultation and site visits. Patients undergo a three-stage counselling procedure prior to starting ART. Drug regimens and monitoring are standardised and prophylaxis against opportunistic infections (isoniazid and cotrimoxazole) is offered routinely. Laboratory and pharmacy services, using named-patient dispensing, are centralized. The programme is designed to ensure that data on clinical and economic outcomes will be available for programme evaluation. Between November 2002-December 2004, ART delivery has been established at 70 ART workplace ART sites. The sites range from 200 to 12000 employees, and from small occupational health clinics and general practitioner rooms to larger hospital clinics. During this period, 2456 patients began ART. Of those on treatment for at least three months, 1728 (78%) have been retained on the programme and only 38 (1.7%) patients have failed the first-line ART regimen. This model for delivery of ART is feasible and successful in an industrial setting. The model may be generalizable to other employment health services in settings of high HIV prevalence, and as a model for implementing ART in other types of health-care settings. PMID- 17129857 TI - HIV vaccine trial preparedness among Spanish-speaking Latinos in the US. AB - Latinos are under-represented in HIV/AIDS medical research in the US. Although they are disproportionately impacted by HIV/AIDS, Latinos may be reluctant to participate in HIV vaccine trials. Three focus groups were conducted with 32 Spanish-speaking Latinos recruited from two community-based healthcare organizations in Los Angeles, California. A qualitative focus group interview guide was developed to explore concerns, motivators and intentions in regard to participation in HIV vaccine trials. Mistrust and fear of government emerged as important themes related to reluctance to participate in an HIV vaccine trial. Specific concerns regarding trial participation included: (1) fear of vaccine induced HIV infection, (2) physical side effects, (3) stigma and (4) false induced HIV-positive test results and their social repercussions. Motivators for enrolling in an HIV vaccine trial included: (1) incentives, (2) convenience of participating in a study, (3) sufficient and appropriate study information, (4) personal benefits and (5) altruism. Interventions to facilitate participation by Latinos in HIV vaccine trials should address mistrust and fear of government sponsored HIV/AIDS medical research, increase access to and convenience of clinical trials, address fear of vaccine-induced infection, combat HIV/AIDS stigma and raise awareness of the relevance of HIV/AIDS to Latino communities. PMID- 17129858 TI - Injecting equipment sharing and perception of HIV and hepatitis risk among injecting drug users in Budapest. AB - In central European states, rates of HIV among injection drug users (IDUs) have been low although Hepatitis C (HCV) infection is widespread. The goal of our study was to assess HIV infection, risk perceptions and injecting equipment sharing among IDUs in Budapest, Hungary. Altogether 150 IDUs were interviewed (121 structured interviews between 1999 and 2000 and 29 ethnographic interviews between 2003 and 2004). The majority of them injected heroin (52% and 79%) and many injected amphetamines (51% and 35%). One person tested positive for HIV. Two thirds (68%) shared injecting equipment (syringes, cookers and filters). Some participants said they shared syringes because they were not carrying them for fear of police harassment and that they reused filters as a backup drug supply. In multivariate analysis, sharing of injecting equipment was associated with higher perceived susceptibility to HIV/AIDS, lower self-efficacy for sterile equipment use, higher motivation to comply with peer pressure to use dirty injecting equipment and with having a criminal record. The high levels of injecting risk-behaviors found in this study are a cause for serious concern. Interventions for HIV-prevention need to address not only sharing syringes but also sharing and reusing of other injecting equipment and drug filters. PMID- 17129859 TI - Factors associated with HIV/AIDS high-risk behaviours among female injection drug users. AB - This study examined factors associated with reporting engaging in HIV/AIDS high risk behaviours at two different time points spaced one year apart for a sample of 185 women who were active injection drug users (IDUs). The high-risk behaviours included injecting drugs in the past six months, having shared needles in the past six months and having engaged in unprotected sexual activity in the past thirty days. Through logistic regression modelling it was identified that living with a spouse at year one was significantly and positively associated with high-risk behaviours at both time points. Being prescribed medications for psychological or emotional problems as well as testing positive for the HIV/AIDS virus were significantly and negatively associated with reporting high-risk behaviours at both time points. These results suggest that spousal relationships may play an important role in HIV/AIDS high-risk behaviours of women drug users. An implication of this study is the need to focus on how spousal relationships and issues such as gender and empowerment should be incorporated into the design and implementation of HIV/AIDS prevention and treatment programs. The benefits of comprehensive mental health diagnosis and treatment services as well as HIV testing in reducing harm to female IDUs and their partners are also discussed. PMID- 17129860 TI - Risk factors and prevalence of HIV infection in people seeking health care in an STI clinic in Brazil. AB - Comprehensive care for people seeking assistance in sexually transmitted infections (STI) clinics provides access to a group of patients in need of HIV prevention interventions and represents appropriate locations for the implementation of these activities. The goal of this study was to determine the risk factors for HIV infection. A profile of patients seeking care in an STI clinic in Vitoria, Brazil; including the motives for this, the risk factors and the prevalence for HIV was carried out. Patients were systematically interviewed regarding demographic information, motivation for seeking assistance, risk exposures, and clinical data. Blood samples were collected to test HIV infection in 562 people enrolled in the study. Prevalence of HIV infection was 6.8 per cent (95 per cent CI 4.7-8.9). Prevention and STI symptoms were the main reasons for seeking care. Variables found to be independently associated with HIV infection were: age (> = 30 years) [OR = 3.24 (95 per cent CI 1.40-7.52)]; seeking medical assistance [OR = 3.88 (95 per cent CI 6.22-40.95)]; and illicit drug use [OR = 14.58 (95 per cent CI 6.11-34.72)]. These results found high prevalence of HIV infection in this population and reinforce the need for ongoing STI prevention activities to further decrease HIV and other STI. PMID- 17129861 TI - AIDS, mobility and commercial sex in Ethiopia: Implications for policy. AB - Since the emergence of the AIDS pandemic in sub-Saharan Africa, male mobility has been highlighted as one of the reasons for the spread of the disease with men employing the services of commercial sex workers while away from home. However, sex workers' mobility and the implications of this for their access to prevention services, has largely been ignored. This paper, based on multi-method qualitative research with 60 young sex workers in two Ethiopian towns, reveals that sex workers are highly mobile, moving in order to attract a wider or different client base, for adventure and to conceal illnesses which might be associated with AIDS. In addition, sex workers are affected by restrictions on their movements, with girls working in bars and red-light areas having little free time to access projects. This paper advocates that policy approaches need to take account of this mobility in three ways: first, by exploring ways for girls to access information and maintain contact with support structures while moving between places of work; second, by building the capacity of sex workers to take greater control over decision-making in their day-to-day lives and third, by developing outreach strategies for taking services into bars and red-light areas. PMID- 17129862 TI - Association of educational attainment with HIV risk in African American active injection drug users. AB - This study explored the association between educational attainment and HIV/AIDS risk among African American active injection drug users (IDUs) in Chicago, US. Using snowball sampling techniques, 813 African American active IDUs were recruited for semi-structured interviewing and HIV counseling, testing and partner notification. Logistic regression examined the relationship between level of education attained (three categories: less than high school; equivalent to high school; and greater than high school) and HIV risk behaviors (12 unsafe sex and drug-related practices) and HIV serostatus (positive or negative). Compared with the reference category (less than high school education), those with education equal to high school were less likely to share water, p = 0.044, OR = 0.70 (95%CI: 0.50-0.99). Compared with the reference category, those with education greater than high school were less likely to receive money for sex, p = 0.048, OR = 0.62 (95%CI: 0.38-0.99); share needles with person having HIV or AIDS, p = 0.015, OR = 0.58 (95%CI: 0.37-0.90); and test positive for HIV, p = 0.027, OR = 0.58 (95%CI: 0.36-0.94). The significant associations found between educational attainment and certain HIV risk behaviors and HIV serostatus have implications for tailoring HIV prevention efforts for less educated African American IDUs. PMID- 17129863 TI - Community attitudes towards individuals living with HIV in rural KwaZulu-Natal, South Africa. AB - Antenatal sero-prevalence rates of 30% and more have been reported in KwaZulu Natal since 1998 and over 50% of all adult deaths in 2000 were due to AIDS. Understanding the changing social and cultural attitudes to AIDS is important in providing contextual information to aid the design of interventions. This paper examines community attitudes towards individuals living with HIV. Eleven focus groups were conducted with men and women. Participants were asked to discuss how people living with HIV were treated within the family and community. The discussions were recorded and transcribed in isiZulu and then translated into English. These were coded using Nud*ist 6 software to identify key themes and sub themes using content analysis. Gender and area differences were investigated. Generally participants expressed positive attitudes to the treatment of AIDS patients and felt that people living with HIV were being cared for within families. However, they reported more negative attitudes to those living with HIV by the general community and suggested these attitudes and acts of discrimination influenced disclosure. Discrimination included physical isolation and symbolism such as referring to them using a 'three finger' gesture. Participants also reported mixed responses to known HIV-positive individuals, ranging from sympathy to a lack of care, on the grounds that the person is certain to die. There are gender differences in terms of the attitudes towards people living with HIV. Compassion and hopelessness seem to be more common among women than men. PMID- 17129864 TI - Why the(y) wait? Key informant understandings of factors contributing to late presentation and poor utilization of HIV health and social care services by African migrants in Britain. AB - The majority of new HIV diagnoses in the UK occur in people with heterosexually acquired HIV infection, the majority of whom are migrant Africans. In the UK HIV positive Africans access HIV services at a later stage of disease than non Africans (Burns et al., 2001; Sinka et al., 2003). Employing purposive sampling techniques, semi-structured interviews were conducted with key informants to identify the key issues affecting utilization of HIV services for Africans in Britain. Considerable agreement about the major issues influencing uptake of HIV services existed amongst the key informants. Respondents felt there was high HIV awareness but this did not translate into perception of individual risk. Home country experience and community mobilization was highly influential on HIV awareness, appreciation of risk, and attitudes to health services. Institutional barriers to care exist; these include lack of cultural understanding, lack of open access or community clinics, failure to integrate care with support organizations, and the inability of many General Practitioners to address HIV effectively. Community involvement should include input to ensure there is: better cultural understanding within the health care system; normalization of the HIV testing process; and a clear message on the effectiveness of therapy. PMID- 17129865 TI - Substance use and HIV risks among male heterosexual and 'money boy' migrants in Shanghai, China. AB - There is a growing awareness that internal migration in China might shift the HIV epidemic by broadening the social and sexual mixing of its population. However, little is known about how drug use/abuse might contribute to the spread of HIV. This qualitative study aims to elucidate factors for preventing substance abuse and HIV among two types of male migrants living in the Shanghai metropolitan area; the general migrant population and so-called 'money boys' (those who engaged in same-sex activities for money). Compared to most male migrants, the 'money boys' had a slightly better economic situation; rarely visited their hometowns; used alcohol less but drugs more; had more knowledge about HIV and sexually transmitted diseases; higher HIV/ STD testing rates and fewer HIV risk behaviors. The general male migrants had more misconceptions about HIV (e.g. the need to pay for HIV testing) than the 'money boys'. However, it was noted that 'money boys' who were new to the enterprise and men who have sex with men but did not engage in commercial sex often lacked HIV knowledge and protective skills. Given the needs of various sub-types of 'migrants', differential approaches to HIV prevention are needed. PMID- 17129866 TI - Mandatory pre-marital HIV testing in Nigeria: the public health and social implications. AB - The prevalence of HIV infection among individuals referred from faith-based organizations (FBOs) in south-eastern Nigeria for mandatory pre-marital HIV screening was determined. Of the total of 319 individuals (148 males, 171 females) screened, 25 (7.8%, 95%CI: 4.9-10.7%) were confirmed HIV-positive, comprising 13 (8.8%, 95%CI: 4.2-13.4%) males and 12 (7%, 95%CI: 3.2-10.8%) females. No significant difference was observed in the association between HIV infection and gender (chi2=0.58, df = 1, P < 0.05). The highest prevalence of HIV infection (8.9%) was recorded among individuals in the 21-30 years age category, while the least HIV infection prevalence (5.3%) was observed among persons above 40 years old. There was no significant difference in the association between HIV infection and age (chi2=0.68, df = 3, P < 0.05). Mandatory pre-marital HIV screening could generate social stigmatization and infringement of the fundamental human rights of infected individuals. Voluntary counselling and confidential HIV testing and especially pre- and post-test counselling as the basis of pre-marital HIV testing are more desirable. Guidelines for the management of test-positive individuals and non-concordant couples and the safeguarding of confidentiality should be developed. Training and capacity building for religious leaders, to appropriately manage social issues associated with HIV/AIDS as it affects their organizations, are recommended. PMID- 17129867 TI - Prevalence and correlates of risky sexual behaviors among injection drug users in Tashkent, Uzbekistan. AB - The objective of this paper is to describe prevalence and correlates of sexual risk behaviors among injection drug users (IDUs) in Tashkent, Uzbekistan. Participants in this cross-sectional study completed a questionnaire detailing sociodemographic, medical and drug and sexual risk behaviors and HIV antibody testing. Of 701 IDUs surveyed, only 20.5% reported consistent condom use, which was more likely for women. Prior sexually-transmitted infection (STI) diagnosis was reported by 36.2% of participants and was associated with early (6 months postoperatively) and no early complications. Thirty per cent of our patients developed an adverse event that could potentially lead to revision of the implanted hardware. In patients with Parkinson's disease most of the complications tend to occur during the first 6 months postoperatively, while in dystonic patients most occur between 12 and 24 months postoperatively. PMID- 17129877 TI - External ventricular drain infection: the effect of a strict protocol on infection rates and a review of the literature. AB - External ventricular drains (EVDs), like any surgically-implanted foreign body, are at risk of infection. We present the results of a completed audit loop following introduction of an evidence-based protocol for their insertion and management. There were two phases over a 2-year period. Phase 1 was a retrospective audit of our EVD infection rate. Phase 2 was a prospective audit of the infection rate subsequent to the introduction of a protocol for the insertion and management of EVDs. In phase 1, the infection rate was 27%. In phase 2, the infection rate was 12%. This was a statistically significant reduction (p < 0.05, Chi-squared test). EVD infection is unfortunately a common clinical problem and associated with potential morbidity and mortality. This study demonstrates that adherence to an evidence-based protocol for their insertion and management is associated with a significant reduction in the infection rate. PMID- 17129878 TI - Is the recommended target of 4 hours from head injury to emergency craniotomy achievable? AB - Guidelines recommend that head-injured patients who require life-saving decompressive surgery should undergo surgery within 4 h. To assess the compliance with this recommendation 100 consecutive head-injured patients admitted to a regional neurosurgical unit (RNU) were studied. Time points from head injury to craniotomy were documented and analysed. Twenty-four patients underwent emergency craniotomy, only one being operated on within 4 h. In this cohort of patients there was no relationship between timing of surgery and outcome. In order to investigate whether it is possible to reduce delays in transportation time, theoretical models were created to determine whether direct transfer to the RNU would be faster by land or air ambulance. PMID- 17129879 TI - Neurosurgical management of intracranial aneurysms following unsuccessful or incomplete endovascular therapy. AB - The publication of the ISAT trial in 2002 has resulted in increasing numbers of patients with aneurysmal subarachnoid haemorrhage undergoing endovascular coiling, as first line treatment for aneurysm occlusion. During the first 2-year period post-ISAT, in which all suitable intracranial aneurysms were coiled first, we had to perform surgery in seven patients out of 54 who underwent initial coiling. The reasons for subsequent surgery were failed coiling, residual or recurrent aneurysm necks and delayed aneurysmal rebleeding despite coiling. We reviewed our surgical experience with the repair of these previously coiled aneurysms to identify unique technical difficulties. In particular, we found that an aneurysm height to neck ratio of less than 2:1 after coiling, and the use of stent-assisted coiling presented formidable challenges to direct clipping. PMID- 17129880 TI - Cavernous malformation of the optic chiasm: case report. AB - We report a 14-year-old boy with cavernous malformation of the optic chiasm. He had a 2-year history of gradually worsening visual disturbance. Computed tomography (CT) and magnetic resonance imaging (MRI) revealed a suprasellar mass, findings compatible with craniopharyngioma. The mass was biopsied and histological examination confirmed cavernous malformation. On the second day after the biopsy, he suffered chiasmal apoplexy due to intratumoural haemorrhage, lost visual acuity and developed a field cut. Cavernous malformations arising from the optic nerve and chiasm are extremely rare; only 29 cases have been reported to date. Most patients manifested acute visual acuity and visual field disturbances. Although MRI findings of cavernous malformations in the brain parenchyma have been reported, MRI findings on the optic nerve and chiasm may not be completely diagnostic. Of the 29 documented patients, 16 underwent total resection of the lesion without exacerbation of their preoperative symptoms; in some cases, resection was complicated by risk of damage to the surrounding neural tissue. As patients may suffer intratumoural haemorrhage after biopsy or partial removal of the lesion, the advisability of surgical treatment of cavernous malformations of the optic nerve and chiasm must be considered carefully. PMID- 17129881 TI - Artefactual computed tomographic middle cerebral artery territory infarct. AB - We report an artefactual middle cerebral artery territory infarct on CT scan. This anomaly is suggested by the discrepancy between the clinical and radiological findings. Awareness of this possibility may avoid unnecessary further investigation or treatment. PMID- 17129882 TI - Recurrent cerebral metastasis from a cardiac myxoma: case report and review of literature. AB - We report the case of a 65-year-old patient who had complete excision of an atrial myxoma and subsequently presented to our unit with recurrent cerebral metastasis. The case demonstrates that cardiac myxoma despite benign histological appearance is capable of metastatic spread. PMID- 17129883 TI - Primary vertebral echinococcosis: four cases reports and review of literature. AB - Primary vertebral echinococcosis has been considered to be a hypothetical and exceptional possibility. It affects bones in 1% of reported cases. Spine is involved in 45-50% of these. The disease preferentially affects the thoracic spine. It begins as a single primary cyst containing daughter cysts. We report four cases of primary vertebral echinococcosis, extending into the extradural space without any other primary site of infection. We conclude that primary vertebral echinococcosis, although rare should be considered in destructive lesions of the vertebrae and spine in regions that the disease is endemic. Advanced imaging studies should be performed to diagnose the disease. Early decompressive surgery of the spine, with chemotherapy, is the treatment of choice for these patients. PMID- 17129884 TI - To assess the ability of MRI to predict consistency of pituitary macroadenomas. AB - We prospectively studied the ability of magnetic resonance imaging (MRI) to predict the consistency of pituitary adenomas. The hypothesis was that firm, fibrous tumours would appear homogeneously hypointense on T2-weighted images. Eighty patients who underwent surgery for pituitary macroadenomas were studied. MR images were evaluated preoperatively by the radiologist. The tumour consistency was reported by the surgeon as soft or firm. There were 68 soft tumours and 12 firm tumours. Of the firm tumours, four (33%) were homogeneously isointense, one (8%) homogeneously hyperintense and seven (59%) were heterogeneous in appearance in T2-weighted images. We conclude that the consistency of pituitary macroadenomas cannot be accurately predicted based on MRI signal intensities. PMID- 17129885 TI - Chronic subdural haematoma in patients with Huntington's disease. AB - We studied the frequency of patients who had chronic subdural haematomas (CSDH) and Huntington's disease (HD) in a 1-year study period. In our department a total of 58 patients with CSDH were treated. Four patients (6.9% of them) had HD. Surgical evacuation of the haematoma was performed in all four cases with the use of a twist drill trepanation without a drainage system. PMID- 17129886 TI - The danger of using metallic plate and screw fixation in the young. PMID- 17129887 TI - Posterior tarsal tunnel syndrome: an unusual unrelated cause of late pain after lumbar spine surgery. PMID- 17129888 TI - Abstracts of the 8th International Conference on Xenon CT and Related Cerebral Blood Flow Techniques: cerebral blood flow and brain metabolic imaging in clinical practice. Cambridge, United Kingdom, 10-12 July 2006. PMID- 17129890 TI - "I intend to donate but ...": non-donors' views of blood donation in the UK. AB - Although only 6% of the eligible United Kingdom population regularly donate blood, reasons for low donation rates remain poorly understood. In a grounded theory investigation, blood donors (n = 23) and non-donors (n = 27) completed a questionnaire that included 15 open-ended items. Semi-structured interviews were conducted with sub-samples of donors (n = 5) and non-donors (n = 7). Analysis of open-ended responses and interview transcripts identified five themes. Three themes, namely pro-social views, helping behaviour and awareness had similar relevance for donors and non-donors. Two themes, anxiety and practical difficulties, were used by non-donors to account for non-donation despite intentions to donate. Although non-donors' anxieties may be difficult to overcome, perceived practical difficulties provide scope for intervention. The removal of perceived barriers to donation offers one way of increasing donation rates. PMID- 17129891 TI - Depressive affect among health care seekers: how it is related to attachment style, emotional disclosure, and health complaints. AB - The present cross-sectional questionnaire survey investigated depressive affect among persons visiting their general practitioner (GP). It examined the impact of a number of factors likely to be associated with depressive affect, including demographic variables, severity of medical condition, and personal factors such as ease in disclosing personal information, and attachment style. Results showed that among the sample of patients (N = 198) visiting their GP, depressive affect was generally quite high. In particular, those reporting more depressive affect were the elderly, those with more severe health problems, more psychological problems, and a lower level of emotional disclosure frequency. As expected, attachment style was also related to lack of well-being among patients: those with a less avoidant but more anxious attachment style reported higher levels of depressive affect. In total, 38% of the variance of depressive affect was predicted by this model. Security of attachment was also related to reasons why patients consulted their GP, with avoidant attachment related to physical problems and anxious attachment to mental problems. Theoretical relevance of the findings and implications for medical help-seeking are discussed. PMID- 17129892 TI - Medication barriers and anti-hypertensive medication adherence: the moderating role of locus of control. AB - Locus of control as a moderator of the relationship between medication barriers (e.g., side-effects, forgetting to take medication, and keeping track of pills) and anti-hypertensive medication adherence was examined. Baseline data were obtained from 588 hypertensive veterans. In general, fewer medication barriers, higher internal locus of control and lower external locus of control was associated with better hypertensive medication adherence. Furthermore, internal locus of control served as a moderator (beta = -.74, p < .01) for the relationship between medication barriers and medication adherence; effect size was large. Decomposition of the interaction revealed that the relationship between medication barriers and medication adherence was strongest when internal control was high (b = -.24, p < .01). Higher internal locus of control was beneficial when barriers to medication adherence are low, but at high perceived barriers, locus of control plays less of a role in medication adherence. Future efforts to improve medication adherence should consider the patient's perceived level of medication barriers in conjunction with their locus of control. PMID- 17129893 TI - Predictors and outcomes of early motherhood in the Australian Longitudinal Study on Women's Health. AB - Early motherhood is identified as a social problem, and having children at an early age is assumed to lead to psychological distress, welfare dependence and socioeconomic disadvantage. Analysis of responses from 9,689 young participants in the Australian Longitudinal Study on Women's Health was used to examine predictors and outcomes of early motherhood in Australia. Survey 1 (1996, aged 18 - 23) and Survey 2 (2000, aged 22 - 27), were used to categorize women as Childless, Existing Mothers (before Survey 1) and New Mothers (became mothers before Survey 2). Multivariate logistic regressions provided comparisons on sociodemographics, gynaecological variables, psychological wellbeing and health behaviours. Survey 1 data show that Existing Mothers experience socioeconomic disadvantages and unhealthy lifestyles. However, those who will go on to become mothers earlier than their peers already experience similar disadvantages. Further, the Survey 2 data show that, when these pre-existing disadvantages are controlled for, the additional deficits experienced by early mothers are relatively minor. Social disadvantage predisposes women to become mothers early, and to adopt unhealthy behaviours. However, young Australian women cope well with the challenges of early motherhood. In the longer term, unhealthy lifestyles and low education may lead to ill health and disadvantage, but early motherhood is not the initiator of this trajectory. PMID- 17129894 TI - Factor structure of the Chinese version of the Geriatric Depression Scale. AB - Depression is common in patients following stroke and has been found to be related to the degree of functional disability, recovery and engagement in rehabilitation. Consequently, screening for depression is crucial in this group in order to facilitate the delivery of appropriate psychological support. The current study sought to determine key psychometric properties of the Chinese version of the Geriatric Depression Scale (GDS) in this group. Three versions of the GDS were evaluated, these being the 30-item original measure (GDS-30), the short-form version comprising 15 items (GDS-SF) and a recently developed innovative four-item version (GDS-4). Confirmatory factor analysis revealed the GDS-30 and GDS-SF to offer an acceptable fit to data suggesting utility of these measures for screening. However, the GDS-4 offered a poor fit to the data, suggesting this measure was an inadequate measure of depression in this clinical group. Further, though GDS-30 and GDS-SF measures revealed good internal consistency, the performance of the GDS-4 was marginal. However, all GDS-derived measures demonstrated excellent convergent and divergent validity. It is concluded that the GDS-30 is a useful and appropriate screening instrument in this group. Further, the GDS-SF shows promise in terms of development as a multidimensional measure of depression that may have predictive and outcome monitoring potential. The psychometric shortcomings of the GDS-4 strongly suggest that this measure is unsuitable for screening in this clinical group. PMID- 17129895 TI - Psychometric properties of the 12-item General Health Questionnaire (GHQ-12) in Chinese women during pregnancy and in the postnatal period. AB - The GHQ-12 is a widely used instrument designed to screen for psychological disturbance (defined as the inability to carry out normal psychological functions and/or the emergence of new distressing phenomena), in a wide variety of clinical groups. The Chinese version of the GHQ-12 is widely used in Hong Kong to screen women during pregnancy and in the postnatal period. The underlying factor structure of the GHQ-12 has been extensively investigated and there is strong evidence to suggest that the instrument is multi-dimensional. The current investigation explored the psychometric properties of this instrument using confirmatory factor analysis, logistic regression and reliability analysis in Chinese women during the third trimester of pregnancy and in the postnatal period. Two- and three-factor models offered superior fit to the data compared to the presumed uni-dimensional structure. However, a four-item sub-scale from an eight-item two-factor model was found to be a significant antenatal predictor of postnatal GHQ-12 assessed caseness. Further evaluation of the clinical utility of this specific two-factor model is recommended. PMID- 17129896 TI - Coping with condom embarrassment. AB - This study assesses the embarrassment associated with purchasing, carrying, storing, using and disposing of condoms. It incorporates coping theory into the investigation of embarrassment by analysing the strategies individuals use to cope with embarrassment during condom purchase. The results of a survey show that individuals are embarrassed at various stages related to condom use. Purchasing condoms elicits the most embarrassment, followed by carrying and disposing, while using and storing are the least embarrassing. To cope with their embarrassment while purchasing condoms, people use multiple cognitive and behavioural coping strategies, with embarrassed people using more strategies. Both embarrassment and the number of strategies used decrease with age and experience. It appears that embarrassment associated with condoms remains a barrier to condom acquisition and consistent condom use, particularly among young adult populations. Coping strategies help individuals to bridge the gap between embarrassment and use. PMID- 17129897 TI - A preliminary psychometric evaluation of the Hospital Anxiety and Depression Scale (HADS) in 963 people living with a spinal cord injury. AB - This study aimed to evaluate the preliminary psychometric properties of the Hospital Anxiety and Depression Scale (HADS) in a community sample of adults living with a spinal cord injury (SCI). A cross-sectional design was used with 963 people living in the community with an SCI. Participants were recruited via three spinal centres in the United Kingdom. They completed the HADS and the Life Satisfaction Questionnaire. Psychometric analyses were conducted for the whole sample, by gender and level of injury. The HADS demonstrated good internal consistency, with promising content validity. Two factors, approximating to anxiety and depression were extracted via factor analysis for the whole sample, by gender and level of injury. In line with existing literature, females scored significantly higher than males on the anxiety subscale. Item 7 ("I can sit at ease and feel relaxed") was found to be a complex item, with a potentially different meaning within this population. This study presents preliminary findings, which support the psychometric integrity of the HADS within an outpatient population with SCI. Items that included potential somatic components revealed a more complex factor loading profile. Recommendations are made to further investigate this measure with amendments to such items, incorporating inpatients and independent measures of anxiety and depression to address validity directly. PMID- 17129898 TI - A pilot investigation of a psychosocial activity course for people with spinal cord injuries. AB - This study investigated the general benefits and impact that sports activity courses organized by Back-Up, a charitable trust, have on quality of life, mood, self-efficacy and perceived manageability. Participants were 35 community-based individuals with spinal cord injuries (SCI). Questionnaires were completed at the start and end of the course. Fourteen participants also completed the follow-up questionnaire. Questionnaires included demographic questions, the Life Satisfaction Questionnaire, the Hospital Anxiety and Depression Scale, a measure of Perceived Manageability, the Generalised Self-Efficacy Scale and four questions about what participants had gained. Results found that participants' satisfaction with leisure, generalized self-efficacy and motivation to engage in activities was significantly increased between the start and end of the courses and anxiety significantly reduced. Such results were in line with participants' responses relating to overall gains from the course. At both the start and end of the course, higher Perceived Manageability scores correlated with greater self efficacy and higher Perceived Manageability scores and self-efficacy were correlated with lower depression and anxiety. The study provides evidence of the benefits of sports participation and teamwork for people with SCI, confirming the results of previous research. PMID- 17129899 TI - The importance of children's illness beliefs: the Children's Illness Perception Questionnaire (CIPQ) as a reliable assessment tool for eczema and asthma. AB - A lack of information about disease in children can lead to erroneous views such as children believing that hospital admittance or the presence of a disease is a punishment for a perceived wrong. There has thus far been no standard tool available to measure children's illness conceptualizations from a Leventhalian framework. Three groups of children with eczema, asthma and eczema and asthma between the ages of 7 and 12 years of age were recruited. Children were given the Children's Illness Perception Questionnaire (CIPQ), a 26-item instrument adapted from the Illness Perception Questionnaire for adults. A Kuder - Richardson 20 test of reliability for dichotomous data was performed allowing an estimate of the internal consistency of the measurement scales. It can be seen that, for all three illness groups, internal consistency is acceptable for the timeline and consequences scale. The cure/control scale, however, was not internally consistent for any illness group. As health professionals, we need to develop the means to further understand how paediatric illness beliefs relate to specific disease types, age and psychosocial factors and the utility of this instrument is discussed within this context. PMID- 17129900 TI - The impact of AIDS/sex education by schools or family doctors on Hong Kong Chinese adolescents. AB - Despite the rapid increases of HIV and sexually transmitted infections (STIs) in East Asia, the sex-related knowledge among Chinese adolescents is limited and the standard of sex education is both variable and inconsistent in Hong Kong. The aim of this study is to review the overall effectiveness of the school-based or doctor-based sexual health education currently provided for Hong Kong adolescents. In 1999 - 2000, a large survey on youth risk behaviours was carried out among 15 - 18-year-old students from 21 secondary schools. Of the 8,039 questionnaires, 377 students (4.7%) admitted sexual experience. Multiple logistic regression showed that students who had received school AIDS education were found to be less likely to have sexual intercourse in the past 3 months (odds ratio, OR 0.5; 95% confidence interval, CI 0.3 - 0.9) or use drugs or alcohol before the previous intercourse (OR 0.5; 95% CI 0.3 - 0.9). They were twice as likely to discuss emotional (95% CI 1.4 - 4.0) or puberty issues (95% CI 1.1 - 3.0). Although not statistically significant, sex education by family doctors showed a similar pattern. This result provides the strongest available evidence that school-based sex education programmes can modify sexual behaviour, which, in turn, reduces risk among Chinese students. PMID- 17129901 TI - Access to HIV care among migrant Africans in Britain. What are the issues? AB - In the UK, substantial numbers of new HIV diagnoses are within migrant communities, especially African communities. Current surveillance shows that despite health promotion efforts and advances in therapy these communities are accessing HIV care late. This paper explores the issues influencing the access and uptake of HIV care by migrant Africans in Britain. Using Kleinman's model of health care systems (Kleinman, 1980) as a theoretical framework, we highlight the importance of placing health within its broader context if we are to make significant improvement to the health of HIV-infected Africans in Britain. PMID- 17129903 TI - The relative influence of facial neoteny and waist-to-hip ratio on judgements of female attractiveness and fecundity. AB - Numerous studies have demonstrated that waist-to-hip ratio (WHR) affects perceptions of female physical attractiveness and fecundity. This study tested the assumption that facial attractiveness explained more variance in overall ratings and fecundity than WHR, when the latter is manipulated within the normal range (0.67 - 0.85). One hundred and sixty-one participants (mean age = 21.5) rated the attractiveness, youthfulness, fertility, healthiness and likelihood of being pregnant for 27 photographs of three females in which facial attractiveness in terms of neoteny (three levels) and WHR (three levels) had been systematically digitally manipulated. Facial attractiveness exerted a significant influence on judgements of attractiveness, youthfulness, fertility and healthiness, whereas WHR only affected likelihood of being pregnant. Results are interpreted in terms of neotenous facial attractiveness providing potential mates with information concerning phenotypic and genetic quality. PMID- 17129904 TI - HIV prevention with persons with mental health problems. AB - The HIV seroprevalence rate among persons with mental health problems (PMHP) is substantially higher than that of the general population in the United States. This study examines the efficacy over 12 months of an HIV prevention program with 99 individuals attending outpatient mental health clinics who were randomly assigned to receive either: (a) a seven-session, small-group intervention of Project LIGHT (Living in Good Health Together); or (b) a one-session video intervention. Regression analyses of data from 87% of the sample interviewed at 1 year follow-up revealed that intervention group membership was associated with significantly fewer sexual risk acts. A significant intervention effect for condom use was found for 72% of the sample who were African American, but not for Latino or Caucasian participants. Results from this study suggest that HIV risk reduction groups such as Project LIGHT may have utility in public mental health care settings. PMID- 17129905 TI - Adherence to HAART: perspectives from clients in treatment support programs. AB - The development of highly active antiretroviral therapy (HAART) has resulted in dramatic decreases in morbidity and mortality for HIV-infected individuals. Because the long-term efficacy of HAART is dependent on near perfect adherence, many providers offer services to support adherence, including education, regimen review, counseling, and reminder tools. Little is known regarding the utilization or efficacy of these services. In this paper, we report on data collected during 32 open-ended interviews conducted with individuals receiving services through the New York State Treatment Adherence Demonstration Project. Participants in the project reported deriving benefits from a broad range of project activities. Educational efforts helped clients to recognize HIV as a manageable, rather than terminal, illness. Information and tools focused on specific regimens provided clients with concrete knowledge and skills. Effective communication with medical providers and pharmacists was facilitated by adherence staff who proactively advocated for client needs. Program-based social support was particularly valuable for many participants, helping them to retain needed motivation and strength. Educational, practical, and supportive services were all considered valuable to clients participating in adherence support programs. A mix of services may best meet the range of needs found among persons taking HAART. PMID- 17129906 TI - Preferences for involvement in medical decision making: comparing British and German views. AB - Despite beneficial outcomes of patients' involvement in medical decision processes, it has been claimed that patients are generally not interested in medical decision making (MDM). Whereas current research focuses on actual MDM, this research explored the impact of nationality and perceptual processes of MDM. Preferences for involvement in decision making were examined in a sample of 204 German and 143 British university students. Hierarchical multiple regressions were employed to explore the link between socio-demographic information, individuals' perceived relationship with their GP, Health Locus of Control - Powerful Others, perceptions regarding the frequency and sufficiency of information provision and involvement in MDM, and individuals' preferred level of involvement and information. A significant amount of the variance in individuals preferences for involvement could be explained (Adjusted R2 = .59, p < .001). Independent t-test analyses showed that British and German perceptions of care differed significantly on a variety of different measures. Separate analyses for the German and British group highlighted cross-national differences in care and preferences for involvement. The study suggests that preferences to become involved might depend more on perceptual processes than actual involvement in decision making, and that communication and national health policy could play an important role. PMID- 17129907 TI - Factor structure and use of the hospital anxiety and depression scale in the homeless and socially marginalized. AB - The homeless have been identified as a group at risk of significant anxiety and depression and screening is therefore highly desirable to identify those needing psychological and psychiatric support. However, the psychometric properties of routine screening instruments have yet to be established in this group. This study sought to determine the psychometric properties of the widely used Hospital Anxiety and Depression Scale in this group. Using a cross-sectional design, 314 clients presenting at homelessness units (shelters) supported by The Salvation Army were administered the HADS. Confirmatory factor analysis revealed three factor models offered the best fit to the data (best-fit model CFI = .98, RMSEA = .06, WRMR = .87). This investigation confirms contemporary research findings that the HADS comprises an underlying tri-dimensional factor structure. However, the internal consistency of the HADS anxiety (alpha = .81) and depression (alpha = .90) sub-scales was excellent. The findings of the current study suggest that the HADS is a suitable screening tool in this group. PMID- 17129908 TI - Psychometric properties of the Chinese version of the diabetes empowerment scale. AB - Metabolic control is central to positive clinical outcome in patients with diabetes. Empowerment has been linked to metabolic control in this clinical group. The current study sought to determine key psychometric properties of the Chinese version of the Diabetes Empowerment Scale (C-DES) and to explore the relationship of the C-DES sub-scales to metabolic control in 189 patients with a diagnosis of diabetes. Confirmatory factor analysis established that the five sub scales of the C-DES offered a highly satisfactory fit to the data. Furthermore, C DES sub-scales were found to have generally acceptable internal consistency and divergent reliability. However, convergent reliability of C-DES sub-scales could not be established against metabolic control. It is concluded that future research needs to address ambiguities in the relationship between empowerment and metabolic control in order to afford patients an evidenced-based treatment package to assure optimal metabolic control. PMID- 17129909 TI - Counteracting stress through leisure coping: a prospective health study. AB - The purpose of this study was to examine stress-buffer or -counteracting effects of leisure coping, by taking into account several key axes of society (i.e., gender, social class, and age) that are essential to characterize the diverse nature of our society. A 1-year prospective survey of a representative sample (n = 938) from an urban Canadian city was conducted. In the total sample, long-term health protective benefits of leisure coping became evident when stress levels were higher than lower (i.e., support for buffer effects of leisure coping). However, a health-protective effect of leisure coping to counteract the impact of stress on health was found substantially stronger for people with lower social class than for those with higher social class. On the other hand, health protective stress-buffer effects of leisure coping were evident regardless of people's gender and age. The findings underscore the importance of giving greater attention to the role of leisure as a means of coping with stress in health practices, particularly among marginalized groups such as individuals with lower social class. PMID- 17129910 TI - Psychological distress, loneliness and disability in old age. AB - With the growth in the number of older people, an increase of mental health problems might be expected. Reducing the amount of suffering and dependence due to poor mental health in old age is a priority requiring a good understanding of the determinants of psychological distress. The objectives were: (a) to measure the prevalence of psychological distress and loneliness in old men and women, living in the community, (b) to clarify the association between psychological distress, health and other explanatory variables and (c) to determine relationship between loneliness and psychological distress. We performed a cross sectional study of 999 people aged 65+. The variables studied were psychological distress (GHQ-12) and self-reported loneliness. The prevalence of psychological distress was 20%. Illness and disability are related to psychological distress in old age; the feeling of loneliness is the single most important predictor of psychological distress, and not knowing neighbours increases the probability of depression. Attempts to improve detection and management of psychological distress in sick older people, in those who appear isolated and express loneliness should be evaluated. PMID- 17129911 TI - A comparison of health behaviours in lonely and non-lonely populations. AB - Loneliness can be defined as perceived social isolation and appears to be a relatively common experience in adults. It carries a significant health risk and has been associated with heart disease, depression and poor recovery after coronary heart surgery. The mechanisms that link loneliness and morbidity are unclear but one of the mechanisms may be through poor health beliefs and behaviours. The aims of this cross-sectional survey of 1289 adults were to investigate differences in health behaviours (smoking, overweight, BMI, sedentary, attitudes towards physical activity) in lonely and non-lonely groups. Lonely individuals were more likely to be smokers and more likely to be overweight - obese. The lonely group had higher body mass index scores controlling for age, annual income, gender, employment and marital status. Logistic regression revealed no differences in sedentary lifestyles. Lonely individuals were significantly less likely to believe it was desirable for them to lose weight by walking for recreation, leisure or transportation. The findings provide support for an association between health behaviours, loneliness and excess morbidity reported in previous studies. PMID- 17129912 TI - A preliminary study into stress in palliative care: optimism, self-efficacy and social support. AB - Caring for the terminally ill is a demanding, but rewarding area of health care. Stressors unique to this working environment--dealing with patient death rather than cure, and supporting entire family units, for example--put caregivers at risk from stress related illness. This study investigated the buffering effects of optimism, self-efficacy and social support against two measures of stress within the palliative setting. Comparisons were made between volunteer (n = 18) and professional (n = 18) caregivers across three caregiving settings. Data were collected retrospectively about personal control variables; use of and satisfaction with, social support; and, perceived general and occupational specific stress levels. Differences in reported stress levels were found both between settings (NHS, hospice and community-based) and between caregiver type (salary status and occupational category). Optimism was more strongly and consistently associated with low levels of perceived stress than was self efficacy. Satisfaction with social support was highest among paid workers, and in terms of buffering stress in the whole sample, appeared mediated by optimism. Worksite interventions targeting caregiver self-efficacy and optimism are recommended as a potential source of stress management within this population. PMID- 17129913 TI - Unexplained chest pain: a review of psychological conceptualizations and treatment efficacy. AB - This review critically considers psychological theories and models used to understand unexplained chest pain, and efficacy of treatment strategies. It discusses the strengths and limitations of current perspectives, and highlights implications for future research and interventions. A comprehensive range of literature examining unexplained chest pain, and published over the last three decades, was thus reviewed finding that, although unexplained chest pain has been examined as a psychological phenomenon for over 100 years, explanatory models have emerged only in the last two decades. Neither psychophysiological nor psychodynamic models have been significantly advanced. Only cognitive-behavioural models have been explicitly derived to explain and manage the condition, and require further refinement to address conceptual and methodological limitations. Studies assessing treatment efficacy suggest cognitive-behavioural therapy as a first-line therapy, but have failed to establish whether the approach is acceptable and effective in routine care. Comprehensive psychological understanding of unexplained chest pain, and its management, is therefore developing but is far from complete. Cognitive-behavioural interventions show promise but are likely to be enhanced by greater theoretical clarity and understanding of resistance to their implementation. PMID- 17129916 TI - The effects of childbirth-related post-traumatic stress disorder on women and their relationships: a qualitative study. AB - There is converging evidence that 1%-2% of women develop post-traumatic stress disorder (PTSD) as a result of childbirth. The current study aimed to explore the long-term effects of childbirth-related PTSD on women, their relationship with their partner and their relationship with their child. Semi-structured interviews were carried out with six women who reported clinically significant PTSD after birth, ranging from 7 months to 18 years beforehand. Interviews were transcribed and analysed using thematic analysis. Childbirth-related PTSD was found to have wide-ranging effects on women and their relationships. Women reported changes in physical well-being, mood and behaviour, social interaction, and fear of childbirth. Women reported negative effects on their relationship with their partner, including sexual dysfunction, disagreements and blame for events of birth. The mother-baby bond was also seriously affected. Nearly all women reported initial feelings of rejection towards the baby but this changed over time. Long-term, women seemed to have either avoidant or anxious attachments with their child. It is concluded that childbirth-related PTSD can have severe and lasting effects on women and their relationships with their partner and children. Further research is needed to compare this to normal difficulties experienced by women after having children. PMID- 17129917 TI - Effects of area and family deprivation on risk factors for teenage pregnancy among 13-15-year-old girls. AB - Information is needed about how the effects of socio-economic deprivation on teenage pregnancy are mediated by proximal risk factors, in order to target area wide and family interventions more effectively. Using a 2x2 factorial design, we tested the separate and interacting effects of area deprivation and family deprivation on six specific proximal risk factors for teenage pregnancy: early sexual activity, life expectations, knowledge and beliefs about contraceptives, attitude to abortion, beliefs about love, and use of local sexual health services. Data were collected from 201 13-15-year-old girls in deprived and non deprived families living in deprived and more affluent areas of the United Kingdom. Area deprivation significantly increased early sexual activity, and both area and family deprivation significantly reduced life expectations. Significant interactions between area and family deprivation showed that the impact of living in a deprived area depends to some extent on family circumstances, with implications for targeting different types of intervention. Living in a deprived area increased early sexual activity much more markedly among girls in deprived families, so interventions to reduce early sexual activity could target individually deprived girls living in deprived areas. Living in a more affluent area increased life expectations, but only among girls in non-deprived families, so both area-wide and individually targeted interventions would be needed to raise life expectations among girls most at risk of teenage pregnancy. PMID- 17129918 TI - Predictors and correlates of coping well with early motherhood in the Australian Longitudinal Study on Women's Health. AB - Women who become mothers at an early age are characterised by socio-economic disadvantage and unhealthy lifestyles; however, some cope extremely well. This paper describes Australian women who become mothers at an early age, in order to identify factors that predict coping. The younger cohort of the Australian Longitudinal Study of Women's Health was used to identify 1064 young women who became mothers between Survey 1 and Survey 2. These women were categorised on the basis of the Mental Health Index of the SF-36 as having High, Normal or Low mental health. Survey 1 data were used to examine predictors, and Survey 2 data to examine correlates, of mental health. Surprisingly, few socio-demographic or health-related variables predict level of coping with early motherhood. Women who would have High mental health as mothers were likely to be in paid work, had few symptoms, and had low levels of stress. They were least likely to have a history of miscarriage and most likely to use contraception. There were no significant effects for other socio-demographic factors, or health-related behaviours. In the longer term, however, all young mothers may suffer an increasing level of disadvantage and distress relative to their peers. PMID- 17129919 TI - Postnatal depression among mothers in the United Arab Emirates: socio-cultural and physical factors. AB - Postnatal depression (PND) has been found to affect women in cultures around the world. This study sought to further identify the prevalence and related socio cultural and physical factors in Arab women from the United Arab Emirates (UAE). The study involved a sample of Emirati women recruited in a government maternity hospital in Abu Dhabi who completed demographic questionnaires soon after giving birth (n=125) and the Edinburgh Postnatal Depression Scale (EPDS) at 3 months (n=86) and 6 months postpartum (n=56). Data are presented in three categories of: No Depression (scores of 0-9), Borderline Depression (scores of 10-12) and Depression (scores of 13+). It was found that at 3 months, this sample had 22% of mothers falling into the Depression category and another 22% falling in the Borderline Depression category. At 6 months, this fell to 12.5% Depression category and 19.6% Borderline Depression category. Relationships between higher depression scores and risk factors included; not breastfeeding, giving birth to the first child, poor self body image and view of weight, poor relationship with mother-in-law, and an older age at marriage. Results are discussed in relation to UAE and Islamic culture. PMID- 17129920 TI - Story-based scales: development and validation of questionnaires to measure subjective health status and cultural adherence in British Bangladeshis with diabetes. AB - Questionnaires that measure subjective health status are increasingly used in clinical trials. But scales based on the quantification of subjective traits ("rate your feelings on a scale of 1 to 5") and initially developed in western population samples may not be valid for use in minority ethnic groups, even if accurately translated. The measurement of cultural adaptation and assimilation in immigrant groups is important for health research but has well documented methodological challenges. The aim of this study was to develop valid and reliable questionnaires to measure subjective health status and cultural adherence in a minority ethnic group, using the story as the unit of inquiry. The design was a multi-phase study involving (a) narrative interview, (b) vignette construction, (c) questionnaire development, and (d) questionnaire validation in relation to two scales (well-being and cultural adherence) in British Bangladeshis with diabetes. Using data from in-depth narrative interviews (i.e., a non-directive research technique in which the participant is invited to "tell me the story about your diabetes, starting with when you first noticed anything wrong", and the only prompts used are "tell me more about that" or "what happened next?"; Greenhalgh, Helman, & Chowdhury, 1998; Muller, 1999), we constructed culturally congruent vignettes to depict different subjective health states and behaviours. We refined these items in focus group interviews and validated the instruments on 98 Bangladeshi participants, randomly sampled from GP diabetes registers in inner London and interviewed by a Bangladeshi anthropologist. We used factor analysis to explore the underlying structure in the responses to questionnaire items, plus Cronbach alpha tests to measure internal consistency of scales. The questionnaires were acceptable and credible to Bangladeshi participants with diabetes. Ninety of 98 participants were able and willing to complete them with interviewer assistance. Following factor analysis, we produced two definitive instruments. The well-being scale was a single-factor model with four story-based items (measuring depression, anxiety, physical energy, and social activities), with a Cronbach's alpha of .92. The cultural adherence scale was a single-factor model with five items (measuring religious restrictions, ethnic practices, and social ties), with a Cronbach's alpha of .83. In conclusion, this study has produced two important outputs: (a) easy-to administer, story-based questionnaires that measure well-being and cultural adherence, which are specific to British Bangladeshis with diabetes; (b) a general method for developing story-based instruments to quantify the subjective experience of illness and adherence to cultural norms, which potentially has applications beyond the study population. PMID- 17129921 TI - Incorporating adolescent females' perceptions of their partners' attitudes toward condoms into a model of female adolescent condom use. AB - The highest rates of sexually transmitted infections in the U.S. occur among adolescent females. One prevention strategy promoted for sexually active adolescents is condom use: therefore, influences on correct and consistent condom use are worth examining. Because interventions and observational research into predicting and increasing condom use have yielded mixed results, we hypothesized that a theoretically driven model incorporating female adolescents' perceptions about partner sentiments along with their own perceptions, intentions, and behaviours would improve condom use predictions. We also measured condom use errors and consistency for a more precise estimate of effective use than is common in the literature. In three structural equation models tested on a sample of 519 female adolescents, we found that intentions were associated with both correct and consistent condom use; that females' expectancy beliefs about condom use were associated with intentions; and that females' expectancy beliefs about partners' sentiments reduced the impact of their expectancy beliefs about condom use. The implications of these relations upon condom use correctness and consistency are discussed with respect to informing interventions, among other future research. PMID- 17129922 TI - The effect of hormone replacement therapy on mood and everyday memory in younger mid-life women. AB - Research on the effect of hormone replacement therapy (HRT) on both mood and memory indicates that oestrogen may enhance verbal memory in younger mid-aged women. This study examined the effect of HRT on everyday memory, while accounting for mood changes, in women between ages 40 and 60. A within-subjects comparison of 17 women, showed that mood, everyday memory, working memory, and delayed verbal memory improved after 3 months of HRT use. The improvement in memory was not mediated by mood, but changes in mood were moderated by exercise habits. The results suggest that verbal memory in particular may be enhanced by HRT in this age group, and everyday memory is an important construct to consider in future research. PMID- 17129923 TI - Impact of physical and psychological resources on functional autonomy in old age. AB - Functional autonomy is a core condition of successful ageing. To maximize autonomous functioning is not only a claim of social policy but also primarily an individual need. Part of the challenge of preventing unnecessary dependence in old age is to recognize the diversity among the elderly and the different availability of their physical, psychological and social resources. The aim of this study is to examine the age- and time-related changes in functional autonomy (activities of daily living, ADL, and instrumental activities of daily living, IADL) and the psychophysical resources correlated with these changes in a sample of 441 healthy elderly persons aged 65-95. Furthermore, we are interested in the long-term predictive power of physical resources (objective and subjective health, physical strength) and psychological resources (memory) on functional autonomy. Results show significant age- and time-related deteriorations in functional autonomy and in most psychophysical resources. Structural equation model analyses were performed to test the long-term predictive power of these resources. Results suggest that ADL is better predicted by physical resources than by psychological ones, but for IADL the reverse is the case; here memory variables play a dominant role. Physical and psychological variables are thus specifically related to different components of functional autonomy. This has to be taken into account in the development of preventive and intervention programmes. PMID- 17129924 TI - Tinnitus Stages of Change Questionnaire: psychometric development and validation. AB - This study describes the development of the Tinnitus Stages of Change Questionnaire (TSOCQ), which aims to assess the readiness of tinnitus patients to change their behaviors and attitudes in relation to tinnitus. Its ability to predict treatment outcome was also investigated. One hundred and fifty-one patients with tinnitus completed the questionnaire. Through repeated principal component analyses, together with judgments of the factors' theoretical relevance, empirically and clinically meaningful scales were derived. A factor solution in accordance with the transtheoretical model (TTM) and with acceptable psychometric properties was found with the five scales of the TSOCQ labeled Precontemplation (Medical Solution), Precontemplation (Helplessness), Contemplation, Preparation and Action/Maintenance. Participants scoring high on Precontemplation (Helplessness) and low on Action/Maintenance had better treatment outcomes. In conclusion, the TSOCQ had some ability to predict treatment success, but further research is required into the utility of the TTM for understanding the rehabilitation process in tinnitus patients. PMID- 17129925 TI - Differential efficacy of a cognitive-behavioral intervention versus pharmacological treatment in the management of fibromyalgic syndrome. AB - Given that studies about the differential efficacy of existing treatments in fibromyalgia syndrome are scarce, the aim of this study was to compare the differential efficacy of a cognitive-behavioral and a pharmacological therapy on fibromyalgia. Using a randomized controlled clinical trial, 28 fibromyalgic patients were assigned to one of following experimental conditions: (a) pharmacological treatment (i.e., cyclobenzaprine), (b) cognitive-behavioral intervention (i.e., stress inoculation training), (c) combined pharmacological and cognitive-behavioral treatment and (d) no treatment. The results show the superiority of cognitive-behavioral intervention to reduce the severity of fibromyalgia both at the end of the treatment and at follow-up. We conclude that cognitive-behavioral interventions must be considered a primary treatment of fibromyalgia syndrome. PMID- 17129926 TI - An examination of the psychometric properties of the Rosenberg Self-Esteem Scale (RSES) in Chinese acute coronary syndrome (ACS) patients. AB - The psychometric properties of the Rosenberg Self-Esteem Scale (RSES) as a clinical research instrument for acute coronary syndrome (ACS) patients were investigated in a translated Chinese version of the instrument. A confirmatory factor analysis was conducted on the RSES to establish its psychometric properties in 128 ACS patients over two observation points (within 1 week and 6 months post-admission for ACS). Internal and test-retest reliability of the RSES TOT (all-items) and RSES-POS sub-scale (positively valenced items) were found to be acceptable. The RSES-NEG sub-scale (negatively valenced items) lacked acceptable internal reliability. The underlying factor structure of the RSES comprised two distinct but related factors, though there was inconsistency in best model fit indices at the 1-week observation point. The use of the RSES as two sub-scales (RSES-POS and RSES-NEG) may be clinically useful in evaluating the influence of this important psychological construct on the health outcomes of patients with ACS. Directions for future research are indicated. PMID- 17129927 TI - Women coping with AIDS in Africa: contributions of a contextually grounded research methodology. AB - The primary objective of this report is to describe a contextually grounded approach to the investigation of coping and psychological functioning in AIDS sick mothers. Five AIDS-sick women on antiretroviral therapy who had at least one child under 6 years of age living in their households were interviewed. Among the issues that emerged was the complexity of the women's psychological functioning, the fact that they made strategic decisions about disclosure aimed at protecting important social support networks, and that their well-being was dynamically influenced by factors in the home and community. The study provides evidence for the value of a contextual approach when investigating the psychological functioning of HIV positive mothers living in poor high-HIV prevalence communities in developing countries. PMID- 17129928 TI - The role of social support in burnout among Dutch medical residents. AB - Burnout levels among medical residents are considered high. A lack of social support has shown to have a direct effect on emotional exhaustion, and depersonalization, two of the three burnout indicators. In this study, we examined the satisfaction of medical residents with social support (emotional, appreciative and informative) received from supervisors, fellow medical residents, nurses and patients. In addition, the correlation between social support and burnout was studied. Medical residents were significantly more dissatisfied with the emotional, appreciative and informative support received from their supervisors compared with fellow residents and nurses (respectively, 13.4+/-4.0 vs. 9.9+/-2.8 and 10.0+/-2.4; 10.0+/-2.9 vs. 7.4+/-2.0 and 7.3+/-1.8; and 7.2+/-2.3 vs. 5.4+/-1.6 and 5.3+/-1.5; p<.001). Significant independent effects were found on emotional exhaustion: from dissatisfaction with emotional support [Beta=.44, p<.001, total R2=.25] and dissatisfaction with appreciative support from supervisors [Beta=.30, p<.01, total R2=.11]. Moreover, dissatisfaction with emotional support from supervisors had an independent significant effect on depersonalization [Beta=.33, p=.001, total R2=.14]. The best predictor of burnout appeared to be dissatisfaction with emotional support received from supervisors. Our results suggest that intervention programs should not only focus on the medical residents, but also on the supervisors to improve their supportive skills. PMID- 17129929 TI - Knowledge and attitudes of Pakistani medical students towards HIV-positive and/or AIDS patients. AB - As the incidence of HIV increases, one of the major steps in preventing a widespread epidemic is to make certain that medical students are prepared to recognize and treat HIV infections and their related conditions, and to counsel patients about avoiding risks that might lead to infections. This cross-sectional study assessed the knowledge level of 357 medical students and their attitudes about AIDS and HIV enrolled in a Medical College in Karachi, Pakistan. Only 6% of the students had complete knowledge on symptoms of HIV/AIDS and 7% of the students had complete knowledge on the modes of transmission of HIV. Statistical analysis of demographic factors affecting knowledge was done. Linear regression and Maentel-Haenszel tests showed that older and clinical students were more knowledgeable of symptoms and modes of transmission of HIV/AIDS. Ten attitudes were correlated with knowledge and none of these showed an association. These results on knowledge indicate that education about HIV/AIDS should be incorporated in the curriculum and interventions must be taken by public health professionals to avoid poor treatment outcomes. PMID- 17129930 TI - Environmental stress, psychological stress and allostatic load. AB - The mechanism by which chronic caregiving stress results in poor health is not well understood. The objective was to determine whether such a mechanism may be allostatic load, a novel concept specifying physiological systems that may suffer cumulative wear and tear following chronic stress, leading collectively to poor health. The study examines the association of allostatic load with environmental and psychological stress in the contexts of dementia caregiving and relinquishment of care, and is a 2-year longitudinal comparison of three groups: 80 new dementia spouse caregivers, 120 veteran caregivers, and 60 non-caregivers. Data comprised allostatic load markers and environmental and psychological stress measures. Cross-lagged analyses produced a statistically significant association between psychological stress and one allostatic load component (primary mediators). Psychological stress was a better predictor of primary mediators than environmental stress. Primary mediators rose with time for caregivers, but not for non-caregivers. A greater rise was evident for caregivers who had relinquished their role by the second year, although the level of psychological stress actually declined. Primary mediators are a key component of the relationship between allostatic load and prior stress. When allostatic load is treated as an outcome of stress, it is important to distinguish environmental and psychological stress. PMID- 17129931 TI - Depression, anxiety, and perceived hassles among entering medical students. AB - This study investigated depression and anxiety among students who were entering medical school prior to the onset of their medical curriculum. Entering students reported financial, day-to-day, academic, and time hassles as concerns. Interestingly, the population characteristics of gender, marital status, and ethnicity impacted the type of self-reported hassles indicated by the students. Measurements of depression and anxiety indicated that entering medical school students' emotional status resembles that of the general population. The results suggest that it is the rigors of the medical curriculum that may play an important role in the increased prevalence of depression and anxiety for students during their medical education. Further, students who are entering medical field already have concerns about medical school and are in the process of anticipating the necessary adjustment to the challenges ahead of them. Preventative programming efforts should begin early in medical education and address a wide variety of concerns from academic, to interpersonal relationships and financial worries. PMID- 17129932 TI - Parental reports of emotional and behavioural difficulties on the SDQ for school age children with vertically acquired HIV infection living in London. AB - This study uses a screening questionnaire (the Strengths and Difficulties Questionnaire, SDQ) to indicate levels of parental or carer concern about behavioural or emotional difficulties in a representative group of school-age children with vertically acquired HIV attending London clinics. Fourteen per cent of the total 107 children screened were reported to have behavioural and emotional difficulties scoring in the "abnormal" range. Older children tended to have higher scores. Overall, levels of reported difficulties were found to be similar to those reported for other chronic childhood illnesses and slightly higher than in the general child population. PMID- 17129933 TI - Attributions of behavior in the pediatric mild closed head injury (CHI) population. AB - Forty-one children between the ages of 6 and 11 years with a history of a mild closed head injury and 23 age-, gender-, and IQ-matched typically developing control children participated. All of the children in the CHI sample were referred for a magnetic resonance imaging (MRI) scan at 3 months post-injury and children in the CHI sample were further divided into MRI-Negative (n=31) and MRI Positive (n=10) conditions according to the MRI results. Parents and teachers completed behavioral checklists at three points, including just prior to the MRI and 6 months post-injury. Prior to the MRI (Time 2), in both samples of children with a CHI, parents reported an increase in externalizing symptoms after the CHI. At 6 months post-injury (Time 3: post-MRI), parents of children in the MRI Positive group did not report any significant changes in their child's externalizing behaviors when compared with pre-MRI, yet parents of children in the MRI-Negative group reported a further increase in externalizing behaviors. Parent attributions also changed as a function of group membership; whereas no changes were noted in the other two groups, parents of children in the MRI Negative sample ascribed more controllability at Time 3 when compared with Time 2. PMID- 17129934 TI - The training & support programme for parents of children with ataxia: a pilot study. AB - The aim of the study was to evaluate the impact of the Training and Support Programme among parents of children with ataxia. A total of 39 parents expressed an interest in the TSP, 27 (mean age 41; range 25-58) returned baseline questionnaires and completed the study. Twenty-four children (mean age 12.5 years, SD=12.4) received the TSP. Data were collected by self-administered questionnaires mailed to parents immediately before attending the TSP and at 4 month follow-up. Interviews were conducted with 10 parents immediately following TSP completion. Comparisons over time revealed significant decreases in parental anxious mood (p=.011), depressed mood (p=.046) and perceived stress (p=.020) and significant improvements in generalized self-efficacy (p=.010), satisfaction with life (p=.045) and parental health status (p=.020). Parents reported improvements in children's mobility, "jumping legs", sleep patterns, energy and activity levels, relaxation, and happiness (e.g., more smiles). Parents felt closer to their children and one parent had become "more accepting" of their child's ataxia. Results indicate that the TSP may enhance the psychosocial well-being of parents of children with ataxia although a randomized controlled trial would be necessary to determine whether the changes reported here are in fact due to attending the TSP. PMID- 17129935 TI - Changes over time among homeless young people in drug dependency, mental illness and their co-morbidity. AB - A cross-national survey was conducted among 358 recently homeless young people in Melbourne and Los Angeles. Drug dependence and mental illness were assessed at baseline, 6 and 12 months. At each time point, participants were classified as no condition, drug dependent, having a mental illness or dual condition. Low levels of drug dependence or mental illness or both were reported at each data point. Most young people were classified as neither drug dependent nor having mental illness at baseline and remained so for the 12-month period and few remained drug dependent, or had continuing mental illness or both. Half to two-thirds of young people classified as having one or both conditions were subsequently reclassified as "no condition". There was no evidence of a consistent pathway from either drug dependence or mental illness to co-morbidity. Most young people reported an absence of mental illness and/or drug dependence at each time point. Improvement from one or both conditions at each transition suggests it may be more effective to provide interventions to address drug dependence and mental health problems to young people early in their experience of homelessness. PMID- 17129936 TI - Psychosocial mediators of group cohesion on physical activity intention of older adults. AB - Considerable evidence has indicated that group-based physical activity may be a promising approach to reducing and preventing age-related illness. However, this research has not examined the mechanisms by which cohesion may impact on behaviour. The purpose of the present research was to utilise the theory of planned behaviour to investigate the mechanism by which group cohesion may affect physical activity intention. Participants were recruited from an existing physical activity intervention studying the effects of group cohesion on physical activity behaviour. The outcomes of this intervention are reported elsewhere. This paper presents data from a sub-sample of the intervention population (N=74) that examined the mediating relationships between the theory of planned behaviour and group cohesion on physical activity intention. Analyses showed that attitude and perceived behavioural control mediated the relationship between specific group cohesion concepts and physical activity intention. The direct measure of subjective norm failed to display a mediating relationship. The mediating relationships displayed between attitude and perceived behavioural control and physical activity intention provide insight into potential mechanisms by which group cohesion may affect behaviour. PMID- 17129937 TI - How does psychological processing relate to compliance behaviour after lung transplantation? A content analytical study. AB - Non-compliance is one of the crucial problems impairing outcome after transplantation. Fourteen lung transplant recipients were interviewed about their thoughts regarding transplant-related topics. Compliance was assessed by doctors. The psychological processing was investigated by content analysis. Highly compliant patients perceived more advantages by transplantation. In contrast, low compliant patients reported either an emotional distance to the lung or a closer relationship to the donor. Furthermore, they showed a contradictory relationship to the medical staff. There are some indications that perception of advantages by transplantation is crucial to compliance. This experience takes place in the context of a good staff-patient relationship. Emotional distance to the lung or nearness to the donor are further contributing factors of non-compliance. PMID- 17129938 TI - Psychological factors and use of antiepileptic drugs: pilot work using an objective measure of adherence. AB - Given the current emphasis on the "concordance" prescribing model, a study was designed to determine the influence of patients' beliefs about epilepsy, beliefs about medication and a range of neuroepilepsy variables on drug adherence among a sample of epilepsy patients. A special feature of the study was the use of a credible objective measure of drug adherence. Psychological health was also assessed. Thirty-seven patients were recruited from a local epilepsy clinic. Beliefs about epilepsy (illness representations), beliefs about epilepsy medication, anxiety, depression, neuroepilepsy status and adherence were all measured. Data were collected via clinical interview and questionnaire methods. Adherence with drug treatment was determined by an objective measure using low dose phenobarbital as an indicator of adherence and, or, measurement of antiepileptic drug levels. Neither illness representations nor beliefs about epilepsy drugs were related to adherence. With the exception of time since last seizure, which was positively related to adherence, neuroepilepsy variables were unrelated to adherence. A number of significant associations between cognitive representations of epilepsy and mood were found. PMID- 17129941 TI - Effects of cadmium and tributyltin on development and reproduction of the non biting midge Chironomus riparius (Diptera): baseline experiments for future multi generation studies. AB - In this study the effects of tributyltin (TBT) and cadmium (Cd) were tested on the freshwater arthropod Chironomus riparius (Diptera) in life-cycle experiments. To this end, the OECD guideline 218 was extended with reproduction relevant parameters (e.g. number of fertile egg masses per female). Based on these reproduction data the number of larvae for next generation and the population growth rate were calculated. Experiments were performed using environmentally relevant concentrations of the test substances in the sediment (nominal ranges: 50-200 micro g Sn/kg and 0.2-2.1 mg Cd/kg, on a dry weight basis). Quartz sand was used as sediment in order to develop a test system for a following research project, focusing on the effects of TBT and Cd as model stressors on C. riparius populations in multi-generation studies. Both model stressors caused significant effects on development and reproduction of C. riparius. Larval mortality proved to be a sensitive parameter and a clear concentration-response relationship was observed with a significant increase (P < 0.001) at the highest TBT concentration and at a Cd concentration of 1.17 mg Cd/kg dw (P < 0.05). The two highest TBT concentrations emergence was significantly (P < 0.05) delayed compared to the solvent control. For Cd a clear prolonged average main emergence interval was observed, but not for TBT. TBT and Cd inhibited also oviposition. These effects resulted in a concentration-depended reduction of the population growth rate. PMID- 17129942 TI - A comparison of greenhouse gas emissions from inputs into farm enterprises in southeast Queensland, Australia. AB - One of the assumptions underlying efforts to convert cropping land, especially marginal crop land, to plantations is that there will be a net reduction in greenhouse gas emissions, with a gas "sink" replacing a high energy system in which the breakdown of biomass is routinely accelerated to prepare for new crops. This research, based on case studies in Kingaroy in southeast Queensland, compares the amount of greenhouse gas (GHGs) emissions from a peanut/maize crop rotation, a pasture system for beef production and a spotted gum (Corymbia citriodora) timber plantation. Three production inputs, fuel, farm machinery and agrochemicals (fertilizer, pesticides and herbicides) are considered. The study extends beyond the farm gate to include packing and transportation and the time period is 30 years. The results suggest that replacing the crops with plantations would indeed reduce emissions but that a pasture system would have even lower net emissions. These findings cast some doubt on the case for farm forestry as a relatively effective means of ameliorating greenhouse gas emissions. PMID- 17129943 TI - Prediction of tropospheric ozone concentrations by using the design system approach. AB - Data on the concentrations of non-methane hydrocarbons (NMHC), nitrogen oxide (NO), nitrogen dioxide (NO2), carbon monoxide (CO), and meteorological parameters (air temperature and solar radiation) were used to predict the concentration of tropospheric ozone using the Design-Ease software. These data were collected on hourly basis over a 12-month period. Sampling of the data was conducted automatically. The effect of the NMHC, NO, NO2,CO, temperature and solar radiation variables in predicting ozone concentrations was examined under two scenarios: (i) when NO is included with the absence of NO2; and (ii) when NO2 is addressed with the absence of NO. The results of these two scenarios were validated against ozone actual data. The predicted concentration of ozone in the second scenario (i.e., when NO2 is addressed) was in better agreement with the real observations. In addition, the paper indicated that statistical models of hourly surface ozone concentrations require interactions and non-linear relationships between predictor variables in order to accurately capture the ozone behavior. PMID- 17129944 TI - Toxicity and biouptake of lead and arsenic by Daphnia pulex. AB - Acute and chronic toxicity studies were conducted on Daphnia pulex using synthetic lead and arsenic water samples. For acute studies, solutions with 0.25, 0.5, 1.0, 2.0, 5.0 mg/L lead and arsenic along with a control were used. The chronic studies were conducted for 21 days using 0.25, 0.5, 1.0 mg/L lead and arsenic solutions along with a control. Results indicated that the LC50 (48 hour) was 4.0 and 3.4 mg/L for lead and arsenic, respectively. Results from chronic studies suggest that the exposure to lead solutions significantly (P < 0.05) impaired the reproduction rates of Daphnia at the 1 mg/L concentration. However, the reproduction rates were enhanced at low concentrations of arsenic (up to 0.5 mg/L). A second chronic study was conducted to confirm this finding. Results from the second study indicated that lead exhibited significantly higher (P < 0.05) toxicity at 0.5 mg/L concentration, while reproduction rates in all concentrations of arsenic solutions were not significantly different from the controls. Metal analysis on exposed Daphnia, following nitric acid digestion procedures, indicated that Daphnia bio-accumulated 75.3-97.2% of the lead added to the experimental containers. This high lead biouptake coupled with the fast growth, high reproduction rates, and short life cycle all suggest that a Daphnia based remediation (growth and partial harvest) may a viable treatment alternative that is worth considering. However, further field studies have to be conducted to verify this alternative. Biouptake or sequestration by Daphnia of arsenic at all tested concentrations was negligible, thereby, suggesting selective uptake or sequestration by daphnia under the tested pH and temperature conditions. PMID- 17129945 TI - Bio-toxicological assays to test water and sediment quality. AB - This study assessed the bio-toxicological assays efficacy in the control of effluent waters deriving from purification plants, such as industrial or wastewater discharge and marine sediment collected from the Adriatic sea on the Italian Coast. The analysis was performed using either Acute Toxicity Test containing bioluminescent bacteria or Algal Growth Inhibitory Test (used on sediments only). Furthermore, samples were also characterized by microbiological parameters to search for indicators of faecal contamination. From the 29 samples collected from effluent waters, 6 showed an inhibition of bacterial luminescence higher than 20% and the analysis of EC50 expressed a strong toxicity for 4 of them, whereas 9 of the remaining 23 samples exhibited a bioluminescence stimulation defined as "hormesi." No samples of marine sediments displayed a reduction of luminescence exceeding 20% with respect to the control. Toxicity tests can represent efficacious tools to detect the presence of pollutants for the preservation of the aquatic system and human health. PMID- 17129946 TI - The effects of container materials and buffer additives on decreasing the iodide concentration in a disposal vault for spent nuclear fuel. AB - To retard the migration of iodine released from a spent fuel after the break of a container, the reducing effects on the concentration of the iodide by container corrosion products and some buffer additives were examined in a solution with bentonite. Iron and copper, and their corrosion products scarcely reduced the iodide concentration. And kaolinite, chalcopyrite, pyrite, copper ore and galena, known as having a sorption property for iodine, did not noticeably sorb the iodide. However, palm active carbon, silver metal and Ag2O lowered the iodide concentration. Especially, Ag2O put into a disposal container would effectively hinder the migration of iodine to the outside of a disposal vault without a great loss if the pore size of the compacted buffer layer is maintained below 1 mu m. PMID- 17129947 TI - A comparison of anion concentration in surficial groundwater sampled from two types of water quality monitoring wells. AB - Groundwater sampling for monitoring the presence and concentration of contaminants can be done using either depth integrated monitoring wells (MW) or depth specific multi-level sampling (MLS) wells. Depth specific multi-level sampling wells (MLS) are cost-effective, easy to install, and provide very detailed information about the vertical gradient in contaminant concentration. In contrast the MW sampling provides information on the presence of contaminant over large representative area. This study was conducted in two 33 ha blocks of a commercial citrus grove (Valencia orange trees on rough lemon rootstock) in a well drained Ashtabula fine sand (hyperthermic, uncoated, Typic Quartzipsamments). The depth to surficial groundwater at monitoring locations varied from 1.4 to 5.6 m, and the lateral groundwater flow rate was approximately 0.08 m d(-1). Anions were measured in groundwater sampled at 3-week intervals from four pair of MW and MLS in each of the two blocks. Since the screened portion of the MW in this study was placed in the top 150 cm of the surficial aquifer, the sampling parts of the MLS within this depth (2nd and 3rd ports) were considered for comparison. The results showed that the concentration of NO(-/3) N, SO(2-/4) and Cl(-) in the MW samples were similar to the mean of the 2nd and 3rd port MLS sample concentrations over a one year period of sampling. Therefore, MLS sampling provides a technique to assess the groundwater quality very similar to that which can be obtainable by MW technique. In addition, the MLS provides useful information on the vertical gradient of solute concentrations thus allows evaluation of the short-term impacts of land management changes on solute concentrations in the very top layer of surficial aquifer in cost-effective manner. PMID- 17129948 TI - Aerosol penetration properties of an electret filter with submicron aerosols with various operating factors. AB - This study was undertaken to determine the effects of using an electret filter on aerosol penetration. Various factors, including particle size (0.05 to 0.5 micro m), aerosol charge state (neutral and single charge), face velocity (0.1, 0.3, 0.5 and 1.0 m/s), and relative humidity (RH 30% and RH 70%), were examined to assess their effects on aerosol collection characteristics. The results presented here demonstrate that the electric fields of the electret and discharged filter were -1.53 x 104 and -1.3 x 102 (V/m). The penetration through the electret filter with singly charged aerosol and neutral aerosol ranged from 0.4% to 13% and 14% to 29%, respectively. According to these results, the coulombic capture force was dominant for the smaller aerosol and the dielectrophoretic capture mechanism was considered important for the larger aerosol. The level of penetration through the electret filter increased with increasing face velocity and relative humidity. The temperature did not affect the penetration through the electret. Furthermore, from the regression analysis conducted during the operating conditions of this work, the aerosol charge was shown to exert the greatest influence on aerosol penetration. PMID- 17129949 TI - Sewage sludge nutrient solubilization using a single-stage microwave treatment. AB - The effects of an advanced oxidation process combining microwave, hydrogen peroxide and acid hydrolysis in a single stage (MW/H2O2/H+ -AOP) on the process efficiency of sewage sludge treatment and nutrient recovery were investigated. At lower temperature regimes (60-80 degrees C), the soluble phosphate was substantially higher in a two-stage process than in a single stage MW/H2O2/H+ AOP process. However, higher soluble phosphate concentration was obtained for single-stage treatment at the higher operating temperature regimes (100-120 degrees C). With the addition of an inorganic acid, a very high yield of soluble phosphate was obtained in the solution at 120 degrees C. In tests with acid addition, soluble ammonia increased as temperature increased. For single stage MW/H2O2/H+ -AOP, maximum soluble ammonia was obtained at 120 degrees C. Significant concentrations of soluble COD were also obtained in this treatment. A threshold temperature of 80 degrees C was observed, at which all of the COD could be solubilized. However, at higher temperatures (100-120 degrees C), further oxidation processes occurred to form carbon dioxide, resulting in decreased amounts of soluble COD in the solution. PMID- 17129950 TI - Influences of polycyclic aromatic hydrocarbons (PAHs) on soil microbial community composition with or without vegetation. AB - Uptake of three polycyclic aromatic hydrocarbons (PAHs), naphthalene (NAP), phenanthrene (PHN), and pyrene (PYR), from soils by ryegrass, white clover and soybean were investigated in an 8-week pot experiment. The microbial phospholipid fatty-acid (PLFA) patterns in PAHs-contaminated soil were analyzed. Contamination with PAHs inhibited the soil microbial activity. In non-vegetated soils, the total PLFA showed 87% reduction on the addition of PAHs after an 8-week incubation compared to the PAH-free soil; the concentrations of NAP, PHN, and PYR in soils showed 19.4%, 25.5% and 24.3% reduction, respectively, due mainly to the evaporative loss and microbial degradation. In vegetated soils, the reduction of added NAP and PHN levels was not related to the plant biomass nor to the total PLFA. However, the reduction in PYR concentrations in soil mediated by soybean growth was much pronounced than those by ryegrass or white clover growth, but thisis not related to the total microbial biomass as revealed by PLFAs. The principal-component analysis (PCA) of the PLFA signatures revealed significant changes in the PLFA pattern in PAH-spiked soils when different vegetations were planted. Using the PLFA as a biomarker, it was found that the Gram-negative bacteria were more sensitive to PAHs than Gram-positive bacteria and fungi. PMID- 17129951 TI - Physicochemical and surface-active properties of biosurfactant produced using molasses by a Pseudomonas aeruginosa mutant. AB - Production of a microbial surfactant was studied by growing Pseudomonas aeruginosa EBN-8 mutant on varying concentrations (on the basis of total sugars) of clarified blackstrap molasses as a sole carbon and energy source with or without auxiliary synthetic nitrogen source in 250 mL shake flasks. The progress of fermentation process was monitored by measuring the production of metabolites, and surface-active and emulsification properties of the cell-free culture broth. The biosurfactant was isolated from the supernatant by acid precipitation followed by solvent extraction. The amount of rhamnolipids produced was determined by the orcinol method. The highest dry cell biomass (1.67 g/L) and rhamnolipid (1.45 g/L) yields were observed, at 96 h of incubation on 2% total sugars-based molasses amended with sodium nitrate (at C:N, 20:1) with the product yield related to dry cell biomass (YP/X, g/g) of 0.869, specific product formation rate (V, h(-1)) of 0.295 and volumetric productivity rate (PV, g/L/h) of 0.015. The surface tension of this culture medium dropped to 28.0 from 50.0 mN/m. PMID- 17129952 TI - Adsorption kinetics of natural dissolved organic matter and its impact on arsenic(V) leachability from arsenic-loaded ferrihydrite and Al-ferrihydrite. AB - The present work was broadly divided into two parts: first, the efficiency of synthetically prepared ferrihydrite and Al-ferrihydrite to adsorb dissolved organic matter (DOM) was tested as a function of time, pH and ionic strength and its effects on specific surface area was found. In the second part, the effect of DOM concentration and solution pH was studied to elucidate the influence of this two parameters on As(V) leachability from As(V) loaded ferrihydrite and Al ferrihydrite. It was found that both pH and ionic strength had significant influence on DOM adsorption. The behavior and magnitude of adsorption was significantly different for both the materials. In general, the efficiency of Al ferrihydrite to adsorb DOM was better compared to ferrihydrite. Results indicated that ligand exchange was the dominant interaction mechanism for DOM adsorption on ferrihydrite and Al-ferrihydrite. The study also suggested a strong potential for DOM to mobilize As(V) from the studied materials. At pH 5.0, DOM concentrations of 2.5 and 5.0 mg C/L had negligible effect on As(V) leachability. With increasing DOM concentration from 10 to 30 mg C/L, increased the As(V) leachability into the solution. Net release from ferrihydrite was up to 1.07 mg/g (5.75%) of the total adsorbed arsenic. The corresponding figure for Al ferrihydrite was 1.47 mg/g (6.30%). As the primary mechanisms for the arsenic release from solid phases we identified two different types of mechanisms i.e., dissolution of the solid phase and competition between arsenic and organic anions for sorption sites. PMID- 17129953 TI - Arsenic concentrations and bacterial contamination in a pilot shallow dugwell program in West Bengal, India. AB - Project Well has developed a pilot self-supporting community-based mitigation program to provide arsenic-safe water to the villagers of North 24 Parganas, West Bengal, India. Shallow concrete dugwells, less than 25 feet deep, that tap into an unconfined aquifer are constructed following stipulated guidelines. The design differs from the traditional dugwell in two major ways: (i) there is a layer of coarse sand in the annular space enveloping the outer wall of the concrete cylinder; and (ii) handpumps are used for water extraction to reduce the potential for bacterial contamination. Monitoring programs for arsenic and coliform bacteria in selected dugwells have been completed. In summer, when the water levels were low, the arsenic concentrations were measured. In 11 wells, measured over three years, the average water arsenic concentration was 29 micro gL-1. Two dugwells had high concentrations of arsenic (average 152 micro gL-1 and 61 micro gL-1), but the remaining nine dugwells had an overall average of 11 micro gL-1. Seasonal variation was assessed in five wells with monthly measurements and there was a direct relationship between increases in arsenic concentrations and decreases in the volume of water in the dugwells in the dry summer season. To control bacterial contamination, sodium hypochlorite solution containing 5% chlorine was applied once a month. In 2005, fecal coliform was undetected in 65% (n = 13) of the dugwells but detected at high levels in 35% (n = 7) of the dugwells. The program clearly reduced exposure to arsenic, but we conclude that further study of increases in arsenic concentrations in the dry season are warranted, as well as assessment of ways to more effectively control bacterial contamination such as more frequent chlorination, perhaps with lower doses on each occasion. PMID- 17129954 TI - Removal of aqueous arsenic using iron attached to immobilized ligands (IMLIGs). AB - The present study describes the synthesis of the iron(III) salt of a commercially available immobilized ligand, IMLIG, Octolig-21 and its use to remove arsenic from aqueous solutions. The synthesis was accomplished by treating an aqueous suspension of Octolig-21 with an equal weight of ferrous sulfate heptahydrate under nitrogen. The ferrous salt was then collected by sieving and allowed to oxidize to the iron (III) salt by exposure to air, and treatment with dilute sodium hydroxide converted the composite to the hydroxide. Using standard test water containing 300 ppb As and column chromatography, reduction of the arsenic concentration to 3 ppb or less in the effluent was achieved, using a pair of columns (4.5 cm id; 1780 mL and 2019 mL, respectively) in tandem. Subsequently, the Fe(III)-Octolig composite was tested for capacity, and it was calculated that with an input of 50 ppb As, it could take up to a year for the effectiveness to be exhausted. PMID- 17129955 TI - Effect of carbon substrate on electron acceptor diauxic lag and anoxic maximum specific growth rate in species with and without periplasmic enzyme. AB - The effect of oxidation state of carbon substrate on the diauxic lag of facultative anaerobic denitrifying bacteria growing aerobically upon switching to anoxic growth was studied. Also studied was the effect on the anoxic maximum specific growth rate. Two pure bacteria cultures were used, Paracoccus pantotrophus, denitrifying bacteria containing a periplasmic nitrate reductase (Nap), and Pseudomonas denitrificans, denitrifying bacteria lacking the periplasmic nitrate reductase. The anoxic maximum specific growth rate of both cultures following a period of aerobic growth with identical dilution up to steady-state was indeed affected by the oxidation state of the carbon, with the most oxidized substrate yielding the highest anoxic maximum specific growth rate. The diauxic lags for Paracoccus pantotrophus were considerably shorter than those for Pseudomonas denitrificans, something expected due to the presence of Nap, an enzyme not affected by aerobiosis. Since the activity of Nap in Paracoccus pantotrophus under aerobic conditions has been shown to increase with the extent of reduction of the carbon substrate, it was also expected that the diauxic lag length for these bacteria would decrease as the reduction state of the carbon substrate increased. This could not be demonstrated, as no significant lags were observed for this species. Pseudomonas denitrificans exhibited a shorter diauxic lag with the more oxidized carbon source. PMID- 17129956 TI - A. Claude (1899-1983), C. de Duve (1917-) and G. E. Palade (1912-): Nobel Prize for discoveries in integrated cell physiology. Clarification of aetiology and pathogenesis of a great number of diseases. PMID- 17129957 TI - Kostmann syndrome or infantile genetic agranulocytosis, part one: celebrating 50 years of clinical and basic research on severe congenital neutropenia. AB - Congenital neutropenia in man was first reported 50 years ago by the Swedish paediatrician Rolf Kostmann. He coined the term "infantile genetic agranulocytosis" for this condition, which is now known as Kostmann syndrome. Recent studies have demonstrated a lack of antibacterial peptides and severe periodontitis in these patients despite recombinant growth factor treatment. Moreover, an increased degree of apoptosis of myeloid progenitor cells in the bone marrow has been shown. CONCLUSION: Future studies should aim to clarify the underlying molecular genetic defect in Kostmann syndrome. PMID- 17129958 TI - Are the symptoms and severity of head injury predictive of clinical findings three months later? AB - AIM: To describe symptoms and functional disabilities 3 mo after a minimal, mild or moderate head injury in children, and to analyse relationships between these and initial symptoms and management documented at the time of the injury. METHODS: The sample consisted of all children (0-15 y) admitted to an emergency department during a 1-mo period with a history of head injury. Data were collected from the medical records at the time of the injury, and questionnaires about symptoms/change and functional problems 3 mo after the injury. RESULTS: Symptoms and functional problems were reported in 35% of the children 3 mo after a head injury. Documented altered consciousness (in children less than 5 y) and disorientation (in children 5 y of age or older) at the emergency department were correlated with changes in behaviour at 3 mo. The initial duration of unconsciousness was not predictive of subsequent behaviour. CONCLUSION: The difficulties in predicting outcomes in head-injured children, especially younger ones, indicate the need for a follow-up appointment in order to identify children with special needs. Further studies are needed regarding the early signs and symptoms in different age groups. PMID- 17129959 TI - Glucose intolerance and insulin resistance in extremely premature newborns, and implications for nutritional management. AB - Glucose intolerance and postnatal growth retardation are commonly seen in low gestation newborns. In contrast to the nutrient compositions of parenteral and enteral nutrition, fetuses at equivalent gestational ages receive much more amino acids (protein) but less glucose and lipids through placental transfer. CONCLUSION: A nutrition regime that simulates placental nutrient delivery would potentially minimize glucose intolerance and facilitate early attainment of normal growth velocity with normal body composition after preterm birth. PMID- 17129960 TI - Preterm birth in Sweden: what are the average lengths of hospital stay and the associated inpatient costs? AB - AIM: To provide estimates of the first-year length of stay and inpatient costs of Swedish infants admitted for neonatal care by week of gestation and by birthweight; and to provide estimates of the length of stay and inpatient costs of delivering mothers during the ante- and postpartum period by week of gestation and birthweight of the infant. METHODS: Population-based registry study covering all live singleton deliveries in Sweden between 1998 and 2001 (n=336 136). First year hospitalizations of infants admitted for neonatal care 0-6 d after birth (n=24 583) were tracked, as were hospitalizations of mothers for whom the date of admission lay+/-1 mo from the date of delivery. Monetary values were assigned to each hospitalization using the Nord-DRG classification system. RESULTS: On average, preterm infants (GA < 37 wk) had first-year lengths of stay roughly four times as long as full-term infants admitted for neonatal care (30 d vs 8 d, p<0.0001). The average first-year length of stay of the extremely immature infants (GA 22-25 wk) was more than six times as long that of infants born at 34 36 wk (108 d vs 17 d, p<0.0001). Mothers delivering preterm had an average length of stay slightly more than twice as long (p<0.0001) as that of mothers of full term infants during the ante- and postpartum period. CONCLUSION: The estimated lengths of stay and costs may serve as reference values for a Swedish setting. PMID- 17129961 TI - Neonatal auditory function and depressed Apgar score: correlation of brainstem auditory response with Apgar score. AB - AIM: To examine the relationship between neonatal auditory function and Apgar score in term infants with depressed Apgar scores. METHODS: Brainstem auditory evoked response (BAER) was recorded on day 3 after birth in term infants who had Apgar scores < or = 7 at 1 and/or 5 min. Half of the infants also had Apgar scores at 10 min, with 17 < or = 7. RESULTS: No BAER variables correlated significantly with 1-min Apgar score. However, wave III and V latencies, and I-V, I-III and III-V intervals correlated significantly and negatively with 5-min Apgar score (p < 0.05-0.01). These BAER variables were significantly longer in infants with 5-min Apgar scores < or = 7 than those > 7. Wave V latency and all intervals also correlated negatively with 10-min Apgar score (p < 0.05-0.01). Compared to normal controls, all latencies were prolonged in infants with depressed Apgar scores (all p < 0.05-0.01). All intervals were also prolonged in those with 5-min scores < or = 7 (p<0.05-0.01). Similar results were found when defining the depression of Apgar score as < or = 6. CONCLUSION: A depressed 5- and/or 10-min Apgar score is an indicator associated with neonatal auditory, mainly central, impairment. Apgar score < or = 7 or 6 at 1 min alone is unlikely to be associated with central impairment. PMID- 17129962 TI - Comparative analysis of neonatal morbidity for vaginal and caesarean section deliveries using hospital charge. AB - AIM: To assess the neonatal morbidity of alternative modes of delivery using economic data. METHODS: Two groups of neonatal morbidity data were extracted according to mode of delivery from inpatient claims on National Health Insurance in Taiwan: uncomplicated vaginal and caesarean section deliveries. Outcome variables included number of infants treated as inpatients, duration of hospital stay, and hospital charge during both the first month and the first year of age. RESULTS: Uncomplicated caesarean section delivery of term infants carried a significant increase either in the first month of life or during the first year after birth in both duration of hospital stay and hospital discharge when compared with uncomplicated vaginal delivery. Neonatal jaundice accounted for the greatest morbidity among term newborns, irrespective of delivery mode, followed by infectious, gastrointestinal and respiratory morbidity. Logistic regression analysis demonstrated that uncomplicated vaginal delivery was associated with skin diseases. Uncomplicated caesarean section delivery was associated with infectious disease and disease of the respiratory, digestive and circulatory systems. CONCLUSION: The findings of this study may provide further information for clinicians and would be an important consideration when advising pregnant women on the preferred route of delivery. PMID- 17129963 TI - Rooming-in for preterm infants: how far should we go? Five-year experience at a tertiary hospital. AB - AIM: To determine the rate of rooming-in among preterm infants born in a tertiary hospital. METHODS: We reviewed the records of all preterm infants born at our hospital during a 5-y period, 2000 to 2004. RESULTS: Of the 18 953 neonates born at our institution during this time, 1356 (7.2%) were <37 wk gestational age. Considering only preterm infants with birthweight > or =1500 g, 806 (74.1%) stayed with their mothers and 282 (25.9%) were admitted to the NICU. Of all the preterm infants that initially stayed with their mothers, 42 (5.2%) needed to be transferred to the NICU. When we stratified these preterm infants according to birthweight, we found that 29% of those <1750 g were transferred to the NICU, compared to only 5% of those > or =1750 g. CONCLUSION: Our study supports the idea that the majority of preterm infants, especially those with birthweight > or =1750 g, can safely remain near their mothers at all times during hospital stay, with both clinical and financial benefits. Neonates with birthweight <1750 g should be evaluated carefully to decide whether rooming-in is the best option. Rooming-in should be encouraged in preterm infants. PMID- 17129964 TI - The role of procalcitonin as a predictor of nosocomial sepsis in preterm infants. AB - AIM: To assess the role of procalcitonin in detecting nosocomial sepsis in preterm infants, after the onset of clinical symptoms. SUBJECTS: 100 preterm infants, 24-36 wk of gestation, were followed from the age of 3 d until discharge. Procalcitonin and C-reactive protein (CRP) levels were measured within 3 d of sepsis workup events. RESULTS: 141 blood samples were drawn from 36 infants during 85 episodes of sepsis workup performed between 4 and 66 d of life. Of these episodes, 51 (60%) were not a result of documented sepsis and thereby served as the negative comparison group. Median procalcitonin levels were higher in the septic group compared with the non-septic group at the time of the sepsis workup (2.7 vs 0.5 ng/ml, p=0.003), at 1-24 h after the sepsis workup (4.6 vs 0.6 ng/ml, p=0.003), and at 25-48 h (6.9 vs 2.0 ng/ml, p=0.016). Using high cutoff levels, both procalcitonin (2.3 ng/ml) and CRP (30 mg/l) had high specificity and positive predictive value (97%, 91% and 96%, 87%, respectively) but low sensitivity (48% and 41%, respectively) to detect sepsis. Areas under the ROC curve for procalcitonin and CRP were 0.74 and 0.73, respectively. CONCLUSION: Procalcitonin >2.3 ng/ml or CRP >30 mg/l indicates a high likelihood for neonatal sepsis, and antibiotic therapy should be continued even in the presence of sterile cultures. PMID- 17129965 TI - Minor neurological dysfunction, cognitive development and somatic development at the age of 3 to 11 years in very-low-birthweight infants with transient periventricular echodensities. AB - AIM: To determine, using strict exclusion criteria, whether transient periventricular echodensities (TPE) in very-low-birthweight infants lead to minor neurological dysfunction and problems in cognitive and somatic development in children without major neurological impairments. METHODS: 23 children with TPE were matched to 23 children without TPE. Exclusion criteria were small for gestational age, microcephaly at birth, diplegia, asphyxia, psychomotor retardation, intraventricular haemorrhage grade III/IV, major surgical interventions and malformations. The Kaufman Assessment Battery for Children, Draw-a-Man Test and neuropaediatric examination were used for evaluation. RESULTS: There were no differences in demographic data, growth and socio-economic status. Significant differences with lower results in the TPE group were found in fine motor skills and in the Draw-a-Man Test. In the Kaufman Assessment Battery for Children, all subscales were below average in the TPE group, except the sequential processing scale. In the control group, all subscales were within the average range. CONCLUSION: By using strict exclusion criteria to eliminate other risk factors for minimal neurological dysfunction and poor cognitive development, we were able to focus on the effect of TPE. TPE seem to affect cognitive development and cause minor neurological dysfunction. PMID- 17129966 TI - Immunogenicity of hepatitis A-inactivated vaccine administered to seronegative infants, and serological follow-up 12 months after second dose. AB - AIM: To evaluate a) the safety and immunogenicity of anti-HAV-inactivated vaccine administered during the first year of life to anti-HAV seronegative babies, and b) the antibody persistence in a low/intermediate endemic area. METHODS: After having obtained informed written consent from mothers, 92 babies were vaccinated at 4 and 10 mo of age. All babies were seronegative at birth and did not present HAV-RNA shedding in three serial stool samples taken at 1, 2 and 3 mo of age. RESULTS: No general side effects (fever > 38 degrees C) were observed. After the first dose of vaccine, 70/82 (85.4%) babies developed anti-HAV > 10 mIU/ml and 36/82 (43.9%) > 20 mIU/ml. After the second dose of vaccine, all babies developed a titre > 20 mIU/ml, and GMT was 877 mIU/ml. After 1 y of follow-up, the decreasing rate was similar to that reported for adult populations. Furthermore, three babies doubled the titre observed 1 mo after the second dose, indicating the possible spread of HAV even in a low/intermediate endemic area. CONCLUSION: Anti-HAV vaccine is safe, immunogenic and able to induce immune memory, and can be integrated into the routine infant immunization schedule during the first year of life. PMID- 17129967 TI - School level at 10 years of age in children who required neonatal intensive care in 1980-1989. AB - School level at age 10 was studied in two cohorts of children who had required neonatal intensive care (NIC): cohort 1, children born 1980-1985 (n=310); and cohort 2, children born 1986-1989 (n=245); and two control groups. More than 80% of all NIC children of both cohorts attended the appropriate mainstream grade 3 or 4; 12.9% of cohort 1 and 6.8% of cohort 2 were in mainstream grade 2. Six per cent of both cohorts received special education. Among very preterm children (23 31 gestational weeks), 73.5% of cohort 1 and 80.3% of cohort 2 attended grades 3 and 4, while 22.9% and 12.1%, respectively, were in grade 2. Assistance (remedial teaching, personal assistant or special teaching group) was given to 42.4% of cohort 1 in the mainstream (grades 2, 3 and 4) and to 38.2% of cohort 2 in the mainstream. In cohort 2, more very preterm girls than matched controls received assistance (p<0.05); no corresponding difference was found in very preterm boys. Most children with congenital malformations received assistance in mainstream education or received special education. CONCLUSION: Most NIC children are in mainstream school classes at age 10. Twelve to 23% of very preterm children are 1 y behind. Many NIC children in the school mainstream need assistance at school, but the proportion of children in mainstream education increases markedly with time. PMID- 17129968 TI - Results of staged palliation for hypoplastic left heart syndrome: a complete population-based series. AB - AIM: To study the outcome of staged palliation for classic hypoplastic left heart syndrome. METHODS: Retrospective chart review. Risk factors for mortality were analysed using Cox's proportional hazard regression modelling. RESULTS: From 1993 to 2004, 55 infants underwent Norwood stage I procedure at a median age of 8 d (range 1-19 d). Hospital survival was 39/55 (71%), and there were six late deaths (before stage II). Birthweight, circulatory arrest time and cardiopulmonary bypass time were independent risk factors for stage I hospital mortality (p=0.029, p=0.001 and p=0.003, respectively). Poor right ventricular function prior to stage I was a significant predictor for interstage mortality (p=0.02). Thirty-two patients underwent bidirectional cavopulmonary anastomosis, at a median age of 6.5 mo (range 2.0-9.5 mo), with seven late deaths. Two patients had a heart transplant after stage II. Total cavopulmonary connection (TCPC) was performed in 13 patients, at a median age of 33 mo (range 21-45 mo), without mortality. Kaplan-Meier survival was 58%, 52% and 45% at 6, 12 and 48 mo, respectively. CONCLUSION: Low birthweight, long time on circulatory arrest and cardiopulmonary bypass were risk factors for stage I mortality. Poor right ventricular function was detrimental to intermediate outcome. PMID- 17129969 TI - Head circumference from birth to age 48 months for infants in Sweden. AB - AIM: To describe the development of head circumference, from birth to age 48 mo, of infants in Sweden. METHODS: Every infant born on the 15th of any month in the 1981 birth cohort, living in Sweden as of 31 December 1989, was studied from birth to age 48 mo (n=3158). By collecting data from child healthcare records and then analysing these data using piecewise linear regression, means and other statistics were produced for every age. Data were analysed for all and with exclusions for low birthweight and those born outside Sweden. RESULTS: From the sample of 3158, data for 3107 infants were collected, making the percentage of missing individuals 1.6%; thus, non-response bias is minimized and high national representativeness is achieved. Summary statistics for head circumference are presented. Comparisons are made to international reference materials, and to previous and current Swedish reference curves. CONCLUSION: For the first time in Sweden, and without selection bias, means and distribution of head circumference measurements are documented longitudinally for a nationally representative sample of infants. PMID- 17129970 TI - Extracurricular sports activity around growth spurt and improved tibial cortical bone properties in late adolescence. AB - AIM: To elucidate whether extracurricular sports activity during rapid growth correlates with improved bone properties in late adolescence, a longitudinal observation was performed among 96 high-school enrollments (46 boys and 50 girls, born in 1981-1982) in metropolitan Tokyo. METHODS: In each year of high school, tibial cortical speed of sound (TCSOS) was measured by quantitative ultrasonometry, and participation in extracurricular sports activity (ECSA) since primary school was examined by structured questionnaire. We calculated the number of years since peak height velocity (ysPHV) based on annual records of height from 6 to 18 y of age to indicate progression of puberty. RESULTS: The increase in TCSOS during high school in boys (32.5 m/s) was significantly greater than that in girls (5.4 m/s). The magnitude of positive association between ysPHV and TCSOS attenuated gradually over time. ECSA in grades 7-9 in boys and in grades 4 6 in girls were significant predictors of TCSOS throughout high school, independent of potential confounders. CONCLUSION: The bone benefits of ECSA around the growth spurt are maintainable in subsequent years. The importance of physical activities that are integrated into the ordinary lifestyle of children and adolescents during this crucial period is emphasized. PMID- 17129971 TI - Normative data for triceps and subscapular skinfold thicknesses of Chinese infants. AB - Skinfold thickness is a validated parameter of subcutaneous fat accumulation. Data on triceps and subscapular skinfold thicknesses were collected from 10,043 (5318 males, 4725 females) singletons at 12 hospitals with gestation 31-42 wk. The LMS method using maximum penalized likelihood was used to perform model fitting of the anthropometric centiles for these parameters. Both triceps and subscapular skinfold thicknesses were greater in girls than in boys. Racial differences were found when comparing with other populations. The triceps subscapular skinfold thicknesses of Chinese infants were significantly lower than those of US white infants but larger than those of African-American infants, Israeli infants, Spanish infants and Indian infants. CONCLUSION: This study provides the first set of references for the triceps and subscapular skinfold thicknesses of Chinese infants by gestation and gender. These data are helpful in determining the nutritional status of infants at birth and in assessing the postnatal growth of Chinese infants. PMID- 17129972 TI - Small and growth-restricted babies: drawing the distinction. AB - The terms "small for gestational age" and "intrauterine growth restriction" have been used interchangeably to denote an in utero growth-restricted neonate. However, the two terms are not synonymous; not all small babies are growth restricted and not all growth-restricted ones are small. Research evidence, extending back to the middle of the last century, reveals that the number of growth-restricted babies who escape attention is not negligible and that the postnatal outcome of these babies is not uneventful. This paper highlights this issue and further discusses the available diagnostic tools for the identification of in utero-restricted neonates, that is, clinical assessment, anthropometric indices and obstetric ultrasound. Each of these tools has strengths and limitations, but, if combined, each could complement the other and help differentiate well-grown babies from those who are growth restricted. CONCLUSION: Identification of growth-restricted neonates is feasible through the integrated use of diagnostic tools. PMID- 17129973 TI - Paediatric metabolic syndrome and associated anthropometric indices: the CASPIAN Study. AB - AIM: To determine the prevalence of paediatric metabolic syndrome (MetS) and its best predictive anthropometric index. METHODS: This national study was conducted among 4811 students (2248 boys and 2563 girls) aged 6-18 y. This is the first study of its kind in Iran and, to our knowledge, in Asia as well. Two definitions were used for the MetS: type A was defined based on criteria analogous to ATP III, and type B was defined according to the cut-offs obtained from NHANES III. Both types A and B define high fasting blood sugar as > 100 mg/dl and systolic/diastolic blood pressure as > 90th percentile. RESULTS: The mean (SD) age of students studied was 12.07+/-3.2 y. MetS type A was seven times more prevalent than type B (14% vs 2%, respectively, p<0.0001), and had no significant gender difference. The most frequent components of both definitions of the MetS were low high-density lipoprotein cholesterol (HDL-C) and high triglyceride (TG). Waist circumference (WC) and waist-to-hip ratio (WHR) had the strongest and weakest associations, respectively, with the MetS. CONCLUSION: Establishment of a uniform set of criteria for the MetS in children is needed. Routine WC measurement in the paediatric population may be clinically useful. PMID- 17129974 TI - Growth references for Turkish children aged 6 to 18 years. AB - AIM: To create up-to-date reference standards for Turkish children, and to compare these with growth standards for US children (CDC 2000 Growth Charts) and with previous local data. SUBJECTS AND METHODS: Height and weight measurements of 1100 boys and 1020 girls were obtained by biannual visits to six schools located in relatively well-off districts of Istanbul city. All children came from well-to do families and all were healthy. All measurements were made by two trained technicians. The LMS method was used in the analyses. RESULTS: Heights of the boys and girls in all age groups were close to the updated 2000 USA growth references and showed an increase from data on Turkish children born 30 y earlier. Weight values were high compared to reference data on US children and to the older data on Turkish children. CONCLUSION: These results indicate that height growth in Turkish school-age children of high socio-economic level conforms to the updated growth data on US children. The data also show a secular upward trend in Turkey. Weight-for-age values indicate an increase in obesity. The results also point to the value of collecting and evaluating local growth data periodically. PMID- 17129975 TI - Does bladder voiding during sleep and wakefulness change the behavioural state of infants? AB - AIM: To evaluate whether bladder voiding in healthy infants is accompanied by body movements or any changes in heart rate (HR), respiratory frequency (RF) or electroencephalogram (EEG) frequency during sleep and during wakefulness. METHODS: Polygraphic recordings were performed on 33 healthy infants (17 female) born at term. The infants' age at study entry was 41+/-10 d, and actual body weight was 4876+/-403 g (mean+/-SD). Bladder voiding was recorded by an adapted enuresis detector connected to the polygraphic computer unit. RESULTS: Awakening was observed in 12 (36%) infants 77+/-9 s before bladder voiding. Twenty-one infants (64%) continued sleeping during bladder voiding. In sleeping infants, bladder voiding occurred during non-REM sleep only, and was accompanied by a cortical arousal. During wakefulness, RF was lower, and HR and EEG frequency were higher, but stayed constant during bladder voiding. CONCLUSION: Our observations demonstrate that bladder voiding in healthy infants during sleep is accompanied by body movements and changes in HR and EEG frequency, indicating cortical arousals, whereas during wakefulness these changes cannot be observed. PMID- 17129976 TI - Quality of life in neurologically healthy children with urinary incontinence. AB - AIM: To bring forward the arguments for active treatment of urine incontinence in otherwise healthy children, a quality-of-life (QoL) study was performed. SUBJECTS AND METHODS: A self-rating QoL questionnaire, child-adjusted and validated, was completed by 120 neurologically healthy children, aged 6-16 y, with urinary incontinence. Another 239 age-matched children made up a control group. The two groups were compared both totally and in age-related subgroups (6-8, 9-12, >12 y) concerning the index for all questions, for universal parts (without questions dealing with incontinence) as well as for specific key domains. RESULTS: The patient group had a significantly lower index than the control group both with and without items related to incontinence (p<0.0001). Social situation, self esteem and self-confidence were most influenced, particularly in the youngest children. Thirty-one children (13%) of the control group reported incontinence and did not score their QoL as good as their continent peers but better than the study patients. CONCLUSION: From the quality-of-life aspects, the study supports active treatment of urinary incontinence in children already at younger ages. PMID- 17129977 TI - Is screening for vesicoureteral reflux mandatory in infants with antenatal renal pelvis dilatation? AB - AIM: To determine whether postnatal ultrasound (US) can guide the use of voiding cystourethrography (VCUG) in infants with antenatally detected renal pelvis dilatation (ARPD). METHODS: 14,000 pregnant women consecutively underwent routine US examination during the second trimester. US examinations later in pregnancy were performed as follow-up of previous anomalies or on obstetrical indications. One hundred and six fetuses with ARPD > or =5 mm were identified. Two postnatal US examinations were performed in the newborns: on the 5th to 7th day and during the 3rd week of life. The findings were considered normal when renal pelvis dilatation (RPD) was < or =7 mm on both US examinations, and no calyceal or ureteric dilatation or signs of renal dysplasia or other anomalies were present. VCUG was done 6 to 8 wk after birth. RESULTS: In 53 of 103 analysable infants, the postnatal ultrasonographic findings were normal. The VCUG was abnormal in three of these 53 infants, all with vesicoureteral reflux (VUR) grade I. Of 50 infants who had abnormal US examinations, six had VUR, four of which were grade IV and V reflux. CONCLUSION: In infants with ARPD who undergo two postnatal US examinations with RPD < or =7 mm and have no other abnormalities, VCUG is unnecessary. PMID- 17129978 TI - Autoantibodies linked to autoimmune polyendocrine syndrome type I are prevalent in Down syndrome. AB - BACKGROUND: Patients with Down syndrome are prone to autoimmune diseases which also occur in the recessive disease autoimmune polyendocrine syndrome type I (APS I). Since this disease is caused by mutations in the gene AIRE on chromosome 21, one might speculate that altered expression of AIRE contributes to autoimmune disease in Down syndrome. AIM: To study the prevalence of 11 well-defined autoantibodies, five of which are specific for APS I, associated with various manifestations of APS I in patients with Down syndrome. METHODS: Sera from 48 patients with Down syndrome were analysed. Autoantibodies against 21-hydroxylase, 17alpha-hydroxylase, side-chain cleavage enzyme, aromatic L-amino acid decarboxylase, cytochrome P4501A2, tyrosine hydroxylase, tryptophan hydroxylase, glutamic acid decarboxylase 65, tyrosine phosphatase IA-2 and transglutaminase were analysed using an immunoprecipitation assay, and thyroid peroxidase autoantibodies were measured using a haemagglutination assay. RESULTS: Seven of 48 patients had elevated titres of autoantibodies: one against 21-hydroxylase, three against aromatic L-amino acid decarboxylase, one against cytochrome P4501A2, one against glutamic acid decarboxylase 65 and one against tyrosine phosphatase IA-2. None of the patients had clinical or laboratory signs of disease coupled to the respective autoantibody. CONCLUSION: Four patients with Down syndrome had autoantibodies hitherto regarded as unique for APS I, which may suggest a dysregulation of AIRE. PMID- 17129979 TI - Analysis of Kawasaki disease showing elevated antibody titres of Yersinia pseudotuberculosis. AB - AIM: To elucidate a clinical difference between patients with Kawasaki disease documented with Yersinia pseudotuberculosis infection and patients with Kawasaki disease without Yersinia pseudotuberculosis infection. PATIENTS AND METHODS: From January 1985 to July 2004, 452 patients were diagnosed with Kawasaki disease. Forty-two patients had elevated antibody titres of Yersinia pseudotuberculosis and/or positive stool culture (Yersinia-positive group). Three hundred and thirty patients had no elevated antibody titres (Yersinia-negative group). We compared the clinical characteristics retrospectively. RESULTS: The age of onset in the Yersinia-positive group (3.05+/-2.20 y) was significantly higher than that in the Yersinia-negative group (2.31+/-2.05 y) (p=0.03). The age-adjusted statistical analysis demonstrated that the incidence of coronary artery lesions (dilatations plus aneurysms) in the Yersinia-positive group (22/42, 52.4%) was significantly higher than in the Yersinia-negative group (105/330, 31.8%) (p=0.001), and the incidence of additional administration of immunoglobulin in the Yersinia-positive group (13/36, 36.1%) was significantly higher than in the Yersinia-negative group (41/256, 16.0%) (p=0.004). CONCLUSION: Our data suggest that Yersinia pseudotuberculosis infection might play a role in the developing mechanism of poor response to therapy and the tendency to develop coronary artery lesions in Kawasaki disease patients. PMID- 17129980 TI - Genetic analyses of thiopurine methyltransferase polymorphisms in Greenlandic and Danish populations. AB - AIM: To determine the frequency of thiopurine methyltransferase (TPMT) low activity alleles in the Greenlandic and Danish populations. METHODS: 142 Greenlandic individuals and 200 Danish blood donors were screened for the TPMT G460A and A719G low-activity alleles. RESULTS: Thiopurine methyltransferase low activity alleles were significantly higher in the Greenlandic compared to the Danish population, being 8.1% (95% CI 4.9-11.3) and 3.5% (95% CI 1.7-5.3) (p<0.01), respectively. Except for one Danish patient with an A719G allele (TPMT*3C), all the aberrant alleles were compound G460A and A719G alleles (TPMT*3A). CONCLUSION: In the Danish population, the incidence of thiopurine methyltransferase low-activity alleles was found to be similar to other Caucasian populations previously described. In contrast, the Greenlandic population showed a significantly higher frequency of thiopurine methyltransferase low-activity alleles. PMID- 17129981 TI - Low-density lipoprotein (LDL) particle size in healthy prepubertal children: the STRIP study. AB - BACKGROUND: Small, dense low-density lipoprotein (LDL) particles (diameter < 255 Angstrom) are highly atherogenic, and in adults they are associated with an elevated risk of coronary heart disease (CHD). AIM: To examine possible contributors to LDL particle size distribution in healthy prepubertal children. METHODS: We determined LDL particle size in 176 7-y-old children by non denaturing polyacrylamide gel electrophoresis. Children were divided into two groups in two ways: by using the median of their average LDL particle diameter (261.2 Angstrom) or by using the LDL peak particle diameter (255 Angstrom) as the cut-off point. RESULTS: Children with smaller LDL particles did not significantly differ from children with larger LDL particles with respect to anthropometric variables, serum triglyceride concentration (in all children < 1.7 mmol/L), apoE phenotype, amount and quality of dietary fat, or child's family history of CHD. Furthermore, LDL particle size did not correlate with any of the anthropometric, lipid or nutrient variables. Children with average LDL particle size below the median had higher serum insulin and lower high-density lipoprotein (HDL) cholesterol and apoA-1 concentrations than those with LDL particle size above the median. CONCLUSION: Our data suggest that many factors related to the development of small, dense LDL particles might have their major impact after the onset of puberty. PMID- 17129982 TI - Determinants of compliance with methylphenidate therapy in children. AB - AIM: To get more insight into factors that influence compliance with the use of methylphenidate. METHODS: 22 pharmacies detected children who used methylphenidate. Their parents were sent a questionnaire on the use of methylphenidate. In a case-control study, the influence of patient- and drug related factors on compliance, as reported by parents, was determined. RESULTS: Parents returned 117 (75%) of 157 questionnaires sent out. Of these, 47.8% of the children missed a dose two times a month or more. Forgetting to take the medication (87.9%) was the most important reason. The case-control study showed that children of 16-18 y were more likely to miss doses. Other patient- and drug related factors did not influence the missing of doses. CONCLUSION: A number of known determinants for compliance do not influence the missing of doses of methylphenidate. With about 80% of the parents reporting that their child missed doses of methylphenidate less than 5 times per month, compliance was quite good for the majority of the children. PMID- 17129983 TI - The availability and accessibility of basic paediatric resuscitation equipment in primary healthcare centres: cause for concern? AB - BACKGROUND: Paediatric emergencies in primary healthcare centres are serious events that occur more commonly than envisaged. However, at present, these centres appear to lack the training and equipment to manage common paediatric emergencies. AIM: To determine the availability and accessibility of basic resuscitation equipment in primary healthcare centres. METHODS: A questionnaire survey of 27 primary healthcare centres within the Nottingham City region determined the availability and accessibility of basic paediatric resuscitation equipment and algorithms. RESULTS: No practice had all 21 basic resuscitation items, with 59% of practices having < or =10 of these items. Only 11% of practices had all seven basic airway and breathing resuscitation items, with 52% of practices having < or =4 items. No practice had all eight basic items for circulation management, with 82% of practices having < or =4 of these items. Only two practices had all six basic drug items, with 85% of practices having < or =3 of these items. Only 26% of practices had algorithms for paediatric basic life support and common emergencies, and only 30% of practices kept their resuscitation equipment together. In the last 5 y, less than a fifth of general practitioners were trained in paediatric resuscitation. CONCLUSION: Primary healthcare centres appear to lack the training and equipment to manage common paediatric emergencies. We recommend standardization of equipment and algorithms, training and assessment of key personnel, and critical incident reporting within primary healthcare centres. PMID- 17129984 TI - The role of maternal depressed mood and behavioural soothing on infant response to routine vaccination. AB - BACKGROUND: It has been hypothesized that maternal characteristics may affect infants' experience of pain during stressful medical procedures. AIM: To investigate the role of maternal depressed mood on infants' response to vaccination, and to determine the effectiveness of different soothing behaviours in reducing infant distress. METHODS: Twenty-eight mothers and their healthy, full-term infants participated in a prospective study. At infant age 2 mo, mothers completed the Edinburgh Postnatal Depression Scale (EPDS) and an adapted version of the Profile of Mood States (POMS); at infant age 4.5 mo, mothers were administered the mood scale and were observed with their infants during routine vaccination. RESULTS: Higher levels of maternal depressed mood were predictive of a stronger infant pain response at routine vaccination. Contingencies derived from sequential analyses revealed that the soothing behaviours most effective in reducing infant distress were holding and face-to-face contact, whereas looking at the child from a distance was significantly associated with an increase in infant distress. CONCLUSION: Our findings suggest that interventions to reduce infant pain during stressful medical procedures may include early detection and referral for maternal postpartum depression as well as the promotion of soothing behaviours involving close physical and emotional contact between mother and baby. PMID- 17129985 TI - Subclinical hyperthyroidism due to a thyrotropin receptor (TSHR) gene mutation (S505R). AB - AIM: To identify the molecular defect by which non-autoimmune subclinical hyperthyroidism was caused in a 6-mo-old infant who presented with weight loss. METHODS: Congenital non-autoimmune hyperthyroidism is caused by activating germline mutations in the thyrotropin receptor (TSHR) gene. Therefore, the TSHR gene was sequenced directly from the patient's genomic DNA. RESULTS: Molecular analysis revealed a heterozygous point mutation (S505R) in the TSHR gene as the underlying defect. CONCLUSION: A constitutively activating mutation in the TSHR gene has to be considered not only in patients with severe congenital non autoimmune hyperthyroidism, but also in children with subclinical non-autoimmune hyperthyroidism. PMID- 17129986 TI - Difficult intubation of a child through laryngeal mask airway with two tracheal tubes. AB - Difficult tracheal intubation occurs infrequently. It is estimated that difficult laryngoscopy occurs in 1-2% of patients. Tracheal intubation of especially small infants can be challenging. When faced with a difficult airway, intubation through a laryngeal mask airway is one method of obtaining a secure airway. Here, we report a 23-mo-old girl with chronic lung disease and severe pneumonia, who was admitted to our paediatric intensive care unit. Since the patient could not be intubated by the standard method, because her larynx was up and forward, she was intubated successfully with a laryngeal mask airway through which two consecutive tracheal tubes were inserted. CONCLUSION: Laryngeal mask airway has an important role as a back-up device in case direct visualization of the larynx is not possible. PMID- 17129987 TI - Sleep behaviour in preterm children from birth to age 10 years: a longitudinal study. AB - AIM: To study clinically relevant aspects of sleep behaviour in preterm children in comparison to term children. METHODS: Longitudinal sleep behaviour data were collected prospectively by structured interviews in 130 preterm and 75 control term children from birth to age 10 y. RESULTS: No significant differences in sleep duration (time in bed per 24 h), bedsharing, night wakings, bedtime resistance and sleep-onset difficulties were found between preterm and term children. CONCLUSION: Sleep behaviour does not differ between preterm and term children from birth to age 10 y, indicating that prematurity or neonatal intensive care experience does not significantly affect sleep in the first 10 y of life. PMID- 17129988 TI - Clarithromycin treatment and QT prolongation in childhood. AB - The effect of clarithromycin on the QT interval was studied in a group of 28 children treated for respiratory tract infections. QTc was measured before and following 24 h of treatment. A modest (average 22 ms, 95% CI 14-30 ms) but significant QTc prolongation (p<0.001) was observed, with seven cases having a QTc >440 ms during treatment (including a single case with QTc >460 ms). CONCLUSION: Serial QTc measurements are necessary for early detection of children at risk for drug-induced arrhythmias. PMID- 17129989 TI - Successful management of primary hypomagnesaemia with high-dose oral magnesium citrate: a case report. AB - We report a 40-d-old female infant who presented to our clinic with afebrile generalized convulsive episodes due to severe hypomagnesaemia with secondary hypocalcaemia. Laboratory investigations revealed mild hypoparathyroidism, which was to return normal following magnesium (Mg) treatment, and normal fractional renal excretion of Mg (0.8%). After the diagnosis of primary hypomagnesaemia was established, the patient was discharged with oral Mg subcarbonate and intramuscular Mg sulphate. On regular follow-up until the age of 4 y, the child was asymptomatic. Mean serum calcium levels remained normal, while mean serum Mg levels remained subnormal. During this 4-y period, oral Mg dose was gradually increased while the doses and frequency of administration of parenteral Mg were decreased. Additionally, oral Mg subcarbonate was switched to Mg citrate because of its side effects. Finally, parenteral Mg was discontinued as the dosage of oral Mg supplementation reached a level of 90 mg/kg/d elemental Mg citrate, without any gastrointestinal side effects. Some screening tests were performed to evaluate the complications of chronic hypomagnesaemia. Bone age, bone densitometry, ECG and renal sonography were all normal. Our patient is now 5 y old and symptom free. CONCLUSION: Treatment with high doses of oral Mg was successful in keeping our patient symptom free and normocalcaemic, but cannot fully normalize serum Mg concentrations. Thus, during the regular follow-up of patients with primary hypomagnesaemia, the main target must be to increase oral Mg supplementation to a dosage which can maintain normocalcaemia rather than normomagnesaemia. PMID- 17129990 TI - Familial idiopathic atrial fibrillation with fetal bradyarrhythmia. AB - A woman presented at 28 wk gestation with fetal bradycardia 50 bpm, which persisted until 42 wk when an asymptomatic male baby was delivered. Electrocardiograph at 3 wk of age documented an incessant atrial fibrillation with slow ventricular response. He continued to be asymptomatic, but on follow-up at 16 y of age, 24-h Holter monitor showed a heart rate of 23 bpm and pauses of up to 6 s when a VVIR programme endocardial pacing system was employed. ECG carried out on his asymptomatic father showed intermittent atrial fibrillation, again with a slow ventricular response. CONCLUSION: Atrial fibrillation is extremely rare in children with normal cardiac structure. Most instances of fetal bradycardia are caused by congenital complete heart block. Other rare causes such as atrial fibrillation with fetal bradycardia need to be considered. This case might be a familial disorder and looks to have a good prognosis. PMID- 17129991 TI - Epilepsy with a de novo missense mutation in the sodium channel a1 subunit: a case report. AB - Most epilepsies are characterized as "idiopathic" because of the lack of a known cause. Nevertheless, recently, there has been significant progress in the molecular genetics of idiopathic epilepsy. Mutations in gene-encoding ion channels were found to be the underlying disorder in all idiopathic epilepsies with a known molecular basis. Missense mutations in the voltage-gated sodium channel a1 subunit gene (SCN1A) were firstly identified in patients with generalized epilepsy with febrile seizures plus additional symptoms (GEFS + ). Subsequently, mutations of SCN1A were also found in patients with severe myoclonic epilepsy of infancy (SMEI) or Dravet syndrome, and in patients with borderline SMEI (SMEB), a milder form of Dravet syndrome. We describe a case of a new missense de novo mutation of SCN1A in a child with the clinical features of borderline SMEI syndrome. PMID- 17129992 TI - A four-year-old girl with cough, haemoptysis and anaemia. PMID- 17129993 TI - Viability and acrosome staining of stallion spermatozoa by Chicago sky blue and Giemsa. AB - A simple trypan blue-neutral red-Giemsa staining procedure for simultaneous evaluation of acrosome, sperm head, and tail membrane integrity and morphology has been used to evaluate equine spermatozoa. Some special characteristics and problems have arisen in evaluating stallion semen. One problem was the differentiation of intact vs. damaged sperm tails primarily in frozen and thawed samples. After freezing and thawing, a high percentage of spermatozoa with an unstained head and stained tail were observed. These cells are considered immotile. Therefore, unambiguous differentiation of intact vs. damaged sperm tail membrane is very important for evaluating semen quality. The aim of our study was to develop a method especially for stallion sperm to distinguish more accurately the different cell types. We compared Chicago sky blue 6B (CSB) to trypan blue (TB) for viability staining. CSB/Giemsa staining showed good repeatability and agreement with TB/Giemsa measurements. For densitometry analysis, individual digital images were taken from smears stained by CSB/Giemsa and by TB/Giemsa. A red-green-blue (RGB) histogram for each area of spermatozoa was drawn. Differences of means of RGB values of live vs. dead tails and separate live vs. dead heads from each photo were used to compare the two staining procedures. CSB produced similar live/dead sperm head differentiation and better tail differentiation. TB can be replaced by CSB and this results in more reliable evaluation. After staining with 0.16% CSB and 4 min fixation, 2-4 h Giemsa staining at 25-40 degrees C is recommended for stallion semen. PMID- 17129994 TI - Cellular immunohistochemical localization of the matricellular protein myocilin in the intervertebral disc. AB - Myocilin is a 55-57-kDa protein that is a member of the olfactomedin protein family. It is expressed in the cornea, sclera and trabecular network of the eye, myelinated peripheral nerves, heart, skeletal muscle, trachea and other tissues. Myocilin binds to a domain of fibronectin, type IV collagen and laminen in the trabecular meshwork of the eye, and its expression is influenced by transforming growth factor beta. Because these extracellular matrix components also are common in the intervertebral disc, the objective of our study was to determine whether the matricellular protein myocilin could be detected in the human or sand rat intervertebral disc using immunohistochemistry and to assess its localization. We investigated 16 specimens of human disc tissue and discs from six sand rats. Three human disc cell cultures grown in three-dimensional culture also were evaluated. Immunocytochemical annulus analysis showed the presence of myocilin within the disc cell cytoplasm in some, but not all, cells. Extracellular matrix in both the human and sand rat disc was negative for myocilin localization. Myocilin is believed to play a role in cell-cell adhesion and/or signaling. Myocilin may have such functions within the disc cell population in a manner similar to tenascin, SPARC and thrombospondin, which are other matricellular proteins recently shown to be present in the disc. PMID- 17129995 TI - Human disc degeneration is associated with increased MMP 7 expression. AB - During intervertebral disc (IVD) degeneration, normal matrix synthesis decreases and degradation of disc matrix increases. A number of proteases that are increased during disc degeneration are thought to be involved in its pathogenesis. Matrix metalloproteinase 7 (MMP 7) (Matrilysin, PUMP-1) is known to cleave the major matrix molecules found within the IVD, i.e., the proteoglycan aggrecan and collagen type II. To date, however, it is not known how its expression changes with degeneration or its exact location. We investigated the localization of MMP 7 in human, histologically graded, nondegenerate, degenerated and prolapsed discs to ascertain whether MMP 7 is up-regulated during disc degeneration. Samples of human IVD tissue were fixed in neutral buffered formalin, embedded in paraffin, and sections stained with hematoxylin and eosin to score the degree of morphological degeneration. Immunohistochemistry was performed to localize MMP 7 in 41 human IVDs with varying degrees of degeneration. We found that the chondrocyte-like cells of the nucleus pulposus and inner annulus fibrosus were MMP 7 immunopositive; little immunopositivity was observed in the outer annulus. Nondegenerate discs showed few immunopositive cells. A significant increase in the proportion of MMP 7 immunopositive cells was seen in the nucleus pulposus of discs classified as showing intermediate levels of degeneration and a further increase was seen in discs with severe degeneration. Prolapsed discs showed more MMP 7 immunopositive cells compared to nondegenerated discs, but fewer than those seen in cases of severe degeneration. PMID- 17129996 TI - A simple and inexpensive method for investigating microbiological, enzymatic, or inorganic catalysis using standard histology and microbiology laboratory equipment: assembly, mass transfer properties, hydrodynamic conditions and evaluation. AB - We introduce a generic, simple, and inexpensive method for performing microbiological, enzymatic, or inorganic catalysis with solids using standard histology and microbiology laboratory equipment. Histology cassettes were used to standardize hydrodynamic conditions and to protect the catalysts and their solid supports. Histology cassettes have the following advantages: they are readily available, inexpensive, solvent and acid resistant, automatable, and the slots in the cassette walls allow liquid to circulate freely. Standard Erlenmeyer flasks were used as reaction vessels. We developed a new camera to observe the movement and position of the histology cassettes as well as the liquid in the Erlenmeyer flasks. The camera produces a stable image of the rotating liquid in the Erlenmeyer flask. This visualization method revealed that in a 250 ml Erlenmeyer flask, stable operating conditions are achieved at a shaking frequency of 300 rpm and a fill volume of 30 ml. In vessels with vertical walls, such as beakers or laboratory bottles, the movement of the histology cassette is not reproducible. Mass transfer characterization using a biological model system and the chemical sulfite-oxidation method revealed that the histology cassette does not influence gas-liquid mass transfer. PMID- 17129997 TI - Histological and biochemical characterization of von Ebner's glands in the Syrian hamster; comparison with rat von Ebner's glands. AB - We report here for the first time a morphological description and observations on some of the secretory proteins of the von Ebner's lingual salivary glands (VEG) of the Syrian hamster. Hamster VEG were macroscopically less distinct, but histologically similar to rat VEG. VEG extracts of hamster and rat were assayed for lipase, alpha-amylase and peroxidase activities. Unlike rat VEG, which is rich in lipase activity, hamster VEG extract had no detectable lipase activity and did not react with antibodies to either rat lingual lipase or human gastric lipase in Western blots. Immunohistochemical reactions with the anti-rat lingual lipase antibody were very weak in hamster VEG and strong in rat VEG. Moderate alpha-amylase enzyme activities and immunohistochemical reactions were demonstrated in both hamster and rat VEG. Peroxidase activity was negligible in the VEG, unlike the high activity in the submandibular glands of both species. An 18 kDa von Ebner's gland protein (VEGP), a member of the lipocalin superfamily of hydrophobic ligandbinding proteins, was abundant in rat VEG, but not detected in hamster VEG. Thus, hamster VEG differs from rat VEG in macroscopic appearance and the absence of lipase and VEGP. It is similar to rat VEG histologically and with regard to the presence of alpha-amylase and absence of peroxidase. PMID- 17129998 TI - Detection of gunshot residues on cadaveric skin using sodium rhodizonate and a counterstain. AB - We report a staining method for cadaveric tissue using sodium rhodizonate as a skin marker for gunshot residues and a counterstain for the surrounding connective tissue. We studied six well preserved subjects who had died of close range gunshot injury. Skin fragments were removed from the bullet entrance hole including both the disrupted area and adjacent macroscopically intact tissue. Because microscopic examination of postmortem material is difficult after histomorphologic alterations already have occurred as a consequence of postmortem tissue changes, it is necessary to use a staining method that, while detecting gunshot residues, can also make skin cell constituents recognizable from both qualitative and quantitative perspectives. Triphenylmethane dyes (acid fuchsin, aniline blue WS, light green SF yellowish, brilliant green and ethyl green) have proven appropriate for the purpose. PMID- 17129999 TI - Fluorescence of GFP in sections of fixed tissue. PMID- 17130000 TI - Nail polish is not suitable for sealing coverslips for fluorescence microscopy, but the bottle and brush are useful. PMID- 17130003 TI - What's so special about adolescence? PMID- 17130004 TI - Attitudes that affect the ability of African American preadolescent girls and their mothers to talk openly about sex. AB - Approximately 50% of new HIV infections in the US occur in teenagers, with African American females making up a disproportionate number. The purpose of this study was to identify factors that could encourage delaying sexual debut. Twenty eight African American mothers and their early adolescent daughters were recruited from four inner-city community centers. Seven separate focus groups were conducted for mothers and daughters using principles of Participatory Action Research. Four codes emerged from the data: social norms regarding sex, males, self-esteem, and family structure. These codes support the importance of an intervention to facilitate sexual communication between a mother and her daughter. PMID- 17130005 TI - Relationships of family functioning, self-esteem, and resourceful coping of Thai adolescents with asthma. AB - Within the context of Rosenbaum's theory of learned resourcefulness, this correlational study examined the relationships among family functioning, self esteem, and resourceful coping in Thai adolescents with asthma. A convenience sample of 132 Thai adolescents (aged 12-17 years) with asthma was recruited from the outpatient asthma clinics of four hospitals in Bangkok. Self-administered questionnaires included an assessment of demographic information and asthma status, the revised Family APGAR, the Rosenberg Self-Esteem Scale, and the Children's Self-Control Scale. Hierarchical multiple regression analysis was used to examine the relationships among variables. Effective family functioning had a significant positive effect on self-esteem (beta = .27, p < .01) and resourceful coping (beta = .30, p < .01), controlling for gender and age. However, self esteem was not significantly correlated with resourceful coping (beta = .15, p = .08). The findings suggest that nursing interventions should take into account the role of family functioning in promoting self-esteem and resourceful coping in Thai adolescents with asthma. Recommendations for future research include replication of the study with a larger sample of adolescents with asthma and with adolescents with other chronic illnesses. PMID- 17130006 TI - Perceptions of mental health among recently immigrated Mexican adolescents. AB - Rates of anxiety, depression, and suicidal ideation are high among Latino adolescents in the U.S., many of whom are immigrants. Immigration during adolescence creates risk factors for mental health problems. The purpose of this study was to explore the health-related perceptions of Mexican-origin immigrant adolescents to inform the design of culturally and developmentally appropriate mental health services. This focused ethnography was guided by Bronfenbrenner's ecological framework and symbolic interactionism. Fourteen adolescents were recruited from two non-health-based community settings. Data from one-to-one semi structured interviews and a visual narrative project were coded and analyzed inductively. Three thematic patterns were identified: "mentally healthy," "mentally unhealthy," and "health promotion." Increased awareness of cultural influences and immigration on Latino adolescents' mental health is needed. Mental health nurses are in a unique position to educate and to influence accessibility of services. PMID- 17130007 TI - Negative thinking: a key factor in depressive symptoms in Thai adolescents. AB - Negative thinking, self-esteem, parental bonding, and everyday stressors are factors related to depressive symptoms in studies conducted in the United States, but they have been rarely explored in Thailand. An understanding of factors influencing depressive symptoms in Thai youth will lead to the development of interventions to decrease depressive symptoms among this age group. The purpose of this study was to examine the effects of parental bonding, everyday stressors, self-esteem, and negative thinking on depressive symptoms among Thai adolescents. A random sample of 812 high school students in Chon Buri, Thailand, participated in the study. The prevalence of depressive symptoms varied from 20-21% depending on the measures used. Negative thinking was the best predictor of depressive symptoms in Thai adolescents. Negative thinking also mediated the effects of parental bonding, everyday stressors, and self-esteem on depressive symptoms. PMID- 17130008 TI - Relationship of adolescent physical and sexual abuse to perimenstrual symptoms (PMS) in adulthood. AB - Research is lacking regarding adolescent physical and sexual abuse and perimenstrual symptoms (PMS). This study examined the relationship between adolescent physical and sexual abuse and PMS in adult women. Secondary data analysis of a longitudinal study of a community sample of 568 women (35% underrepresented ethnicities), using the database "Nursing Assessment of PMS: Neurometric Indices," was performed. History of both adolescent physical abuse and sexual abuse was significantly associated with PMS in adulthood. Women with a history of adolescent physical and sexual abuse had significantly more severe PMS patterns with more dysphoria than women without abuse. PMID- 17130009 TI - When breastfeeding may be a threat to adolescent mothers. AB - Sexually abused girls are at risk for premature motherhood. The adolescents who become mothers often do not breastfeed. This review examines the literature and discusses the mental health consequences of childhood sexual abuse that may influence the feeding decisions of adolescent mothers. Adolescents may be reluctant to breastfeed because it may trigger anxiety and lead to discomfort with intimacy, and emotional distancing from their infants. Nurses need to be sensitive to potential sexual abuse histories among adolescent mothers. They should provide balanced and unbiased information to their adolescent patients about feeding methods and encourage adolescents to select the methods best for them. PMID- 17130010 TI - College students' perspective on smoking cessation: "If the message doesn't speak to me, I don't hear it". AB - The purpose of this study was to document the smoking cessation experiences of three groups of 18-24-year-old college students and explore their ideas for cessation. Focus group interviews were conducted with former smokers (n = 7), smokers struggling to quit (n = 7), and smokers with no desire to quit (n = 5). Participants indicated a need for smoking cessation programs that would help the students understand the immediate physiological impact of smoking, addictive nature of nicotine, and benefits of using nicotine replacement therapy. They believed emotional and cognitive strategies for coping with cravings and environmental cues to smoking should be included in smoking cessation programs. Students indicated that cost and convenience were important considerations for smoking cessation programs. They stated that the media messages were for teens and older adults and did not relate to them as college students or young adults. PMID- 17130011 TI - Prescribing update: why choose duloxetine for depression? PMID- 17130012 TI - Cultural competence: what is it? PMID- 17130013 TI - "It's more than a shoe shine--you're touching lives". PMID- 17130015 TI - Male contraception: past, present and future. AB - Male contraception research has yielded a number of promising leads over the past 50 years. Yet, little is known by the public due to lack of institutional support and funding. This is unfortunate since, apart from condom and vasectomy, there are many male methods which may be safer, more effective and easier to use. This paper explores male contraception which has been used in the past and the present and discusses some of its potential developments. PMID- 17130016 TI - Placental dysfunction: pathophysiology and clinical considerations. PMID- 17130017 TI - A review of methodological quality of systematic reviews on multiple pregnancies. AB - We set out to determine the quality of existing systematic reviews on multiple pregnancies. We conducted an electronic search in MEDLINE (1951 - 2005), EMBASE (1974 - 2005) and the Cochrane Database for Systematic reviews (2005:2) and a hand-search of reference lists without any language restrictions to identify relevant reviews. Two reviewers independently selected review articles in which a publicly available database was searched for studies concerning multiple pregnancies, and assessed them for quality of methods of review. Information was extracted on framing of question, literature search and data synthesis. Of 342 citations 14 (4%) eligible reviews were identified. Only 8/14 reviews specified the review question. Adequate literature search without language restriction and the use of a reference list was found in 7/14 reviews, but the risk of missing studies was assessed in only 1/14 reviews. Quality assessment of included studies was reported in 7/14 and tabulation of their findings was reported in 8/14 reviews, but heterogeneity of results was evaluated in only 4/14 reviews. Meta analysis was employed in 3/14 reviews. Systematic reviews of existing studies on multiple pregnancies are infrequent and it is difficult to generate robust inferences from them as they lack good methodology. PMID- 17130018 TI - The 30 minutes decision-to-delivery interval for 'urgent' caesarean sections: an elusive target. AB - The aim of this audit was to evaluate whether the '30 minute decision-to-delivery interval' for 'urgent' caesarean section has shown consistent improvement with repeated audit within this unit. The audit dataset comprised a random sample of all urgent caesarean sections carried out in 2004 classified as 'urgent', i.e. to be completed in 30 min. Nearly one-third of caesarean sections recorded on the delivery suite database as 'urgent' were incorrectly coded. A personal review of case notes was undertaken to ensure accurate data capture. Delivery suite data was analysed by post-hoc modelling of a 'normal' (Gaussian) distribution. The proportion of true 'urgent' caesarean sections completed in 30 min was 50% and some 90% of women were delivered within 40 min. The data were normally distributed, with non-random events, accounted for 25% of the variability. A model for 'urgent' caesarean section, accommodating random and non-random factors closely matched the audit data. We conclude that non-random, institutional, factors reflecting overall delivery suite activity adversely effect the decision to-delivery interval regardless of the performance of personnel and processes within a delivery suite. PMID- 17130019 TI - Pre-term pre-labour rupture of membranes: effect of chorioamnionitis on overall neonatal outcome. AB - Our objective was to evaluate the association between clinical chorioamnionitis following preterm pre-labour rupture of membranes (PPROM) and adverse neonatal outcome. We compared retrospectively, adverse neonatal outcome of singleton pregnancies with documented PPROM who developed chorioamnionitis (cases) with those who did not (controls). Our result showed that poor neonatal outcome was significantly associated with chorioamnionitis (34% vs 13%; p = 0.008). This association was found on multiple logistic regression analysis to be independent (p < 0.05) of other risk factors for poor neonatal outcome, viz: latency period (p = 0.002) and gestational age at delivery (p < 0.001). We conclude that chorioamnionitis complicating PPROM worsen neonatal outcome. The implication of this on expectant management of PPROM is discussed. PMID- 17130020 TI - How useful is external cephalic version in clinical practice? AB - We reviewed retrospectively all women with a singleton breech presentation after 36 weeks' gestation who were referred for an external cephalic version (ECV) in the 3 years from 2001 - 2003. The number (n = 107) referred was low and the overall success rate of 24.5% was also low. Over the 3 years, our practice of ECV prevented only 15 caesarean sections and thus made little contribution to reducing the hospital's overall caesarean section rate. This audit highlights the need for a review of current practices in our hospital. ECV is recommended by the guidelines of the RCOG and it is a concern that the results achieved by enthusiasts of ECV may not be achieved in everyday obstetric practice. PMID- 17130021 TI - Blood transfusion and caesarean section in a developing country. AB - Our objective was to review blood transfusion practices during caesarean section in a developing country. An audit of 463 consecutive caesarean sections and blood transfusions over a 3-year period (2000 - 2002) was undertaken. The data were collected from the records department in a pre-designed proforma and analysed, using EPI - info Statistical Software version 6. A total of 117 out of 463 (25.2%) caesarean section cases were transfused. The rate of blood transfusion for the various indications were as follows: malpresentation (excluding breech), four out of six (66.7%); placenta praevia, 28 out of 49 (59.1%); uterine rupture, five out of nine (55.6%); breech delivery, eight out of 25 (32%); obstructed labour, 35 out of 124 (28.2%); precious baby, one out of four (25%); previous caesarean section, 24 out of 141 (17.0%); severe pre-eclampsia, five out of 45 (11.1%); fetal distress, three out of 28 (10.7%); and others, three out of 29 (10.3%). A total of 78 (67.2%) of caesarean section cases were emergency. A blood transfusion rate of 25.2% during caesarean section is high. The indications for the caesarean section, preoperative anaemia and quantity of blood loss during caesarean section were significant risk factor for blood transfusion. Efforts should be made to reduce the blood transfusion without increasing maternal morbidity and mortality. This is very important because of rising HIV infection in developing country and blood-borne disease. PMID- 17130022 TI - Pregnancy outcome following use of large doses of vitamin B6 in the first trimester. AB - Vitamin B6 is often prescribed for the treatment of nausea and vomiting of pregnancy (NVP), at much higher doses than initially recommended. Large doses of vitamin B6 have been associated with cases of neuropathy. We set out to assess whether higher than standard doses of vitamin B6 during the first trimester of pregnancy were associated with a risk of maternal adverse events, major malformations, miscarriages or low birth weight. This was a prospective comparative observational study. The study group included women who were exposed to >50 mg/day of vitamin B6 during the first trimester; the control group included pregnant women with a non-teratogen exposure. A total of 192 pregnancies were followed-up. The mean dose of B6 used in the study group was 132.3 mg/day (median 110 mg/day, range 50 - 510 mg/day), for a mean period of 9 +/- 4.2 weeks. In this group (n = 96), there were 91 live births, one major malformation and the mean birth weight was 3,542 +/- 512 g. There were no statistical differences in the study endpoints between the vitamin B6 and the control groups. Within the limits of our sample size, higher than standard doses of vitamin B6 do not appear to be associated with an increased risk for major malformations. PMID- 17130023 TI - Understanding operative intervention in childbirth: a patient perspective. AB - A study was undertaken to ascertain patients' understanding of the operative interventions in labour and to assess follow-up by the operator. A total of 200 consecutive women who had undergone caesarean section or instrumental delivery were selected. These women were questioned postoperatively. Questions were asked to ascertain the patients' understanding of the procedure and to assess follow-up by the operator. Seven patients had a forceps delivery, 64 had a ventouse delivery and 129 had a caesarean section. The majority of patients felt that the reason for the operative delivery had been explained to them at delivery and that they fully understood the need for this intervention. A total of 26 women were not seen postoperatively by the doctor who delivered them. Women who underwent forceps or ventouse delivery were less likely to be seen post-delivery, although this difference did not reach statistical significance. PMID- 17130024 TI - An evaluation of the effect of parity and age on malaria parasitaemia in pregnancy. AB - A higher prevalence of malaria infection (peripheral or placental) has been reported in the primigravidae and secondigravidae when compared with multigravidae. This study set out to determine the effect of parity and age on the prevalence of malaria parasitaemia in pregnancy at the booking antenatal visit at the Nnamdi Azikiwe University Teaching Hospital, Nnewi. Peripheral blood smears were examined in 420 pregnant women at their booking antenatal visit and in 200 control subjects attending the outpatient clinic on the same day for malaria parasites. These subjects (pregnant women and controls) met the inclusion criteria of being HIV sero-negative, not sickle-cell positive, did not have a history of recent blood transfusion and had been resident in Nnewi for 1 year. The result showed that there was a statistically significant difference between the prevalence rate of malaria parasitaemia in the primigravidae (87.9%: 109 of 124) and grand multigravidae (63.6%: 28 of 44); and the rates were found to decrease with increasing parity. The primigravidae had a higher mean parasite density (2,155/micro l) when compared with the multigravidae (1,950/micro l). This study also revealed that pregnant women <20 years had the highest prevalence rate of 86.4% (19 of 22). This study demonstrates the higher prevalence of malaria parasitaemia in pregnant women of lower parity, i.e. primigravidae and secondigravidae. Therefore, targeting malaria control efforts to women in their first and second pregnancy will be an important strategy to reach most infected women and minimise resource expenditure. These women should be motivated to use insecticide treated bed nets (ITBN) and other personal malarial control measures during pregnancy. PMID- 17130025 TI - Effect of inter-twin delivery interval on neonatal haemoglobin concentration. AB - The aim of this study was to examine the effect of the inter-twin delivery interval on neonatal haemoglobin concentration. We retrospectively analysed 108 twin deliveries over a 5-year period. Chorionicity was determined by first trimester ultrasound or placental histopathology. The mode of delivery, time of delivery and cord/neonatal blood counts were recorded. A total of 88 sets of dichorionic and 20 sets of monochorionic twins were studied. There was no correlation between inter-twin delivery interval and haemoglobin difference (rho = 0.020, p = 0.857). In monochorionic twin pregnancies, there was a trend towards increasing inter-twin hemoglobin differences with prolonged delivery intervals. However, this trend did not reach statistical significance (rho = -0.303, p = 0.193). In monochorionic twins, there is a trend towards a lower haemoglobin concentration in the twin delivered second. This haemoglobin deficit appears to be related to the inter-twin delivery interval. PMID- 17130026 TI - Haematological profile of healthy pregnant women in Ibadan, south-western Nigeria. AB - There is a dearth of information on the reference values for haematological indices particularly according to the relevant trimesters of pregnant women in Nigeria. The objective of this study was to provide reference values for Nigerian pregnant women. The study took place at the Adeoyo Maternity Hospital and the University College Hospital, both in Ibadan. This descriptive study was carried out over a period of 8 months. Subjects were apparently healthy pregnant women that satisfied the inclusion and exclusion criteria. The mean values (and 95% confidence intervals, CI) of haematological indices were as follows -- First trimester: Haemoglobin (Hb) 112.44 (101.64 - 123.25) g/l, haematocrit (hct) 35 (32 - 38)%, WBC 5.488 (4.025 - 6.950) x 10(9)/l and platelet counts 227.56 (165.21 - 289.90) x 10(9)/l;Second trimester: Hb 100.39 (97.85 - 102.92) g/l, hct 29.3 (28.5 - 30.1)%, WBC 6.57 (6.19 - 6.95) x 10(9)/l, platelet count 229.56 (211.86 - 247.26); and the Third trimester: Hb 98.06 (96.12 - 100.00) g/l, hct 29.4 (28.7 - 29.9)%, WBC 6.92 (6.53 - 7.30), platelet count 186.52 (177.67 - 195.38) x 10(9)/l. These results were compared with those of 52 non-pregnant age matched women volunteers as controls whose mean haematological indices and 95% CI were: Hb 120.51 (116.61 - 124.41) g/l, hct 36 (25 - 48)%, WBC 5.28 (2.9 - 8.7) x 10(9), platelet count 330.87 (176 - 538) x 10(9)/l. The following haematological indices: WBC, platelet counts, RBC, PCT, and PDW, of women between the trimesters showed statistical significance (p value < 0.001 in each case). The WBC is inversely proportional to the PCT and the MCV in the pregnant women was slightly raised. In this study, pregnancy is characterised by lowest values of haemoglobin parameters in trimester three and there are statistically significant differences between the WBC, platelet counts, RBC, PCT, and PDW of women between the three trimesters. PMID- 17130027 TI - Risk factors for hepatitis B infection during pregnancy in a Nigerian obstetric population. AB - We aimed to assess the risk factors for hepatitis B infection among pregnant women in Enugu, southeast Nigeria. This was a prospective case-control study of risk factors for hepatitis B surface antigen positivity among pregnant women seen in two tertiary health institutions in Enugu, southeast Nigeria. It was carried out over an 8-month period, January - August 2005. Of the 1,499 pregnant women attending the antenatal clinic, 4.6% (n = 69) were seropositive for hepatitis B surface antigen. A total of 35 (50.7%) HBsAg positive women and 71 negative controls were interviewed. The risk factors for HBsAg were present in 71% (n = 25) of the positive cases, while only 27% (n = 19) of the negative cases had risk factors. The significant risk factors for HBsAg positivity were higher mean parity, higher number of sexual partners since sexual debut, polygamy and previous positive history of sexually transmitted diseases (STD) (p < 0.05). The two groups did not differ with respect to presence of tattoo or scarification marks, positive history of jaundice or contact with a jaundiced patient, previous blood transfusion or contact with blood products, intravenous drug abuse or sharing personal instruments (p > 0.05). Screening pregnant women for hepatitis B infection on the basis of presence of risk factors may not effective. Universal antenatal screening for HBsAg, health education aimed at reducing risk factors and immunisation of all newborn and those at risk of hepatitis B is advocated. PMID- 17130028 TI - Sociodemographic factors in anaemia in pregnancy at booking in Lagos, Nigeria. AB - Anaemia in pregnancy is a common problem in many developing countries. In Nigeria, it is responsible for 11.0% of maternal deaths. This study examined the sociodemographic factors associated with anaemia in pregnancy. Anaemia was defined as a haemoglobin (Hb) level <11 g/dl. Proportions were compared using the chi2 test. A total of 35.3% of the 374 women studied were anaemic. Anaemia was more common in primigravidae than in parous women of parity 2 - 4 (chi2 = 6.87; p = 0.0087). Low socioeconomic status, (chi2 = 24.67, p = 0.00090), short interval between pregnancies (chi2 = 20.34, p = 0.000144), booking in the third trimester (chi2 = 15.4, p = 0.00045) and recent febrile illness (chi2 = 6.80; p = 0.0091) were associated with anaemia. Women who only used haematenics were more anaemic than those who also used anti-malarial chemo-prophylaxis (chi2 = 57.32, p = 0.00001). Anaemia in pregnancy is associated with several social and demographic factors. To reduce the prevalence of anaemia women need to have a good formal education, be economically empowered and good antenatal care must be made available, accessible and affordable to all women. PMID- 17130029 TI - Vaginal hysterectomy: results and complications of 886 patients. AB - The aim of this study was to determine the feasibility, results and complications of vaginal hysterectomy. A total of 886 consecutive patients who had undergone vaginal hysterectomy for benign gynecological diseases were retrospectively analysed. Vaginal hysterectomy was successfully performed in 96.1% of the nulliparous and 99.9% of the parous patients. The mean duration (min) of the operation was 89.1+/-29.1. The operation time (min) of the nulliparous women was significantly higher than that of the primiparous and multiparous women (109.3+/ 40.2 vs 81.1+/-33.2 and 85.1+/-28.3, respectively). The overall complication rate was 14.6%. The intraoperative and postoperative complication rates were 4.1% and 10.5%, respectively. The most common intraoperative complication was bladder injury (2.5%). Vaginal hysterectomy for benign gynaecological diseases has high feasibility with acceptable complication rates. PMID- 17130030 TI - Probability of early pregnancy loss in women with vaginal bleeding and a singleton live fetus at ultrasound scan. AB - Bleeding is a common feature of early pregnancy affecting about one-fifth of pregnant women in the first trimester. The chance of miscarriage after bleeding and a live fetus at scan has not previously been defined precisely. The purpose of this study was to evaluate the outcome of early pregnancies with a viable singleton fetus that had been complicated by bleeding. A prospective study was performed on 370 women with a singleton live fetus who had presented to the early pregnancy assessment clinic (EPAC) with vaginal bleeding. Women were grouped into light, moderate and heavy loss according to the self-assessed degree of vaginal bleeding. The women were also categorised according to the presence or absence of an intrauterine haematoma. The overall spontaneous miscarriage rate in the study was 11.1%; almost 90% of pregnancies continued to viability. Women with moderate or heavy bleeding had more than twice the rate of miscarriage compared with those with light bleeding. A total of 14% of the women had an intrauterine haematoma and those women were 2.6 times more likely to miscarry than those without (23% vs 9%). This relationship appeared to hold true even after controlling for blood loss. The data presented can be used to guide women with a live fetus about the chance of miscarriage after an episode of vaginal bleeding. We propose that a large multi-centre study should be undertaken to define precisely the risk miscarriage for each gestational week according to a range of clinical and ultrasound characteristics. PMID- 17130031 TI - The effect of diclofenac and paracetamol on pregnancy and implantation rates in infertile women undergoing IVF treatment. AB - We set out to assess the effect of diclofenac on implantation rates, when administered as analgesia following transvaginal oocyte recovery. This was a prospective study of infertile women undergoing IVF treatment in a University Hospital. Subjects (n = 74) were divided in two groups (A and B). Group A (n = 38) received 1 g paracetamol and 100 mg diclofenac and group B (n = 36) received 1 g paracetamol only. All the medication was administered rectally immediately after the oocyte retrieval. Pregnancy and implantation rates were compared between group A and B using the chi2 test. In groups A and B, the implantation rates were 12.4% and 9.6% (p = 0.5) and the pregnancy rates were 28.9% and 19.4%, respectively (p = 0.67). Neither pregnancy nor implantation rates differed significantly between the two groups. Administration of diclofenac to patients at the time of egg collection does not appear to affect implantation or pregnancy rates, while it could be effective in reducing discomfort and pain associated with oocyte retrieval. PMID- 17130032 TI - Saline sonohysterosalpingographic findings in infertile Nigerian women. AB - This paper describes the findings in the first 100 women who underwent saline sonohysterosalpingography (SHG) at two privately owned health facilities in Enugu, South East Nigeria. This was a prospective study of the first 100 consecutive women presenting with infertility to the authors at Mbanefo Hospital and Hansa Clinics, both in Enugu, South-East geopolitical zone of Nigeria from 1 May 2005 to 20 January 2006. Saline sonohysterosalpingography was carried out in the standard way in these women. The findings were analysed using simple percentages and means +/- SD. The procedure was completed in 98 women, while in two others it was not possible to cannulate the uterine cavity. A total of 61 of the women had normal uterine musculature, 37 had interstitial fibroids and two had sonographic features of adenomyosis. Regarding the endometrial cavity, 93 women had a normal endometrium, four had a submucosal fibroid, one had intrauterine adhesions, while the endometrial cavity could not be assessed in two women who could not be cannulated. No case of submucosal polyp or uterine septa or other congenital uterine anomalies was seen. As assessed sonologically, 77 of the study subjects had bilateral patent tubes, while five had unilateral tubal patency. In one woman, there was uncertainty about tubal patency or blockage; in two women, the tubes could not be assessed because of non-cannulation of the uterine cavity and in 15 women, both tubes were blocked. A total of 74 women had normal ovaries; 15 had polycystic ovaries; five had atrophic ovaries consistent with ovarian failure and six women had ovarian cysts. In 18 women, the findings at sonosalpingography (SSG) were confirmed at laparoscopy in 11 women or laparotomy (two women) or by the fact that the patients became pregnant (five women). In 15 (83.3%) of these 18 women, the findings at SSG and laparotomy/laparoscopy or of the woman becoming pregnant were compatible. SSG is a useful screening test for assessing endometrial, tubal and ovarian factors in infertile Nigerian women, thereby obviating the need for laparoscopy and hysteroscopy in the majority of cases. PMID- 17130033 TI - Termination of pregnancy in South Tyneside. AB - We evaluated prospectively, compliance with the Royal College of Obstetricians and Gynaecologists (RCOG) guidelines on termination of pregnancy (TOP) in a cohort of 340 women referred for termination of pregnancy in 2003 at South Tyneside Foundation Trust. The number of referrals represented one-fifth of all births in our unit during the study period. Teenagers were the largest single group of women requesting termination of pregnancy and the majority were nulliparous. There were 85 women who were seeking a repeat termination of pregnancy. The RCOG minimum referral standard was met in 80% of cases. A good number of women were unsure of their menstrual dates and only 5% had used emergency contraception. A total of 96% were either not using contraception, using condoms or taking oral contraceptives irregularly. A total of 50% of the women attended hospital without a Certificate A being completed by the referring practitioner. Surgical termination was preferred over medical termination in the cohort of women who could exercise a choice. It is possible to comply with the RCOG Termination of Pregnancy guidelines to a large extent in a District General Hospital, with some innovation. Close liaison between General Practitioners, Family Planning Clinics and Acute Hospitals is required. The adoption of agreed referral requirements and pathways would help in the delivery of a high quality service as advocated by the guideline. PMID- 17130034 TI - Juxtacervical vesicovaginal fistulae: outcome by route of repair. AB - Vesicovaginal fistula is a major public health problem in Nigeria with diverse medical, psychological and social consequences for the patient. This study compared the outcome of vaginal vs abdominal repair of juxtacervical vesicovaginal fistulae. It was a retrospective review undertaken at the University of Nigeria Teaching Hospital, Aghaeze Hospital and Mbanefo Hospital, all in Enugu, Nigeria, from 1 January 1992 to 31 December 2004. The outcome measures were primary repair success rate, blood transfusion, postoperative urinary tract infection rate and duration of hospital stay. Abdominal repair of juxtacervical vesicovaginal fistula was associated with a significantly higher need for blood transfusion when compared with vaginal repair. Both routes of repair had similar primary repair success rates, postoperative urinary tract infection rates and duration of hospital stay. It was concluded that the route of repair of juxtacervical vesicovaginal fistula should be determined by accessibility of the fistula and whenever possible, the vaginal route should be preferred. PMID- 17130035 TI - The prognostic importance of proliferative activity and oestrogen receptor expression in stage I endometrial carcinomas. AB - The purpose of this study was to evaluate the prognostic significance of steroid hormone receptor proliferation index in endometrial adenocarcinoma. In this study, the correlation between oestrogen receptor expression, proliferation index and FIGO grade, age, myometrial invasion, tumour size and menopause status was evaluated in 40 patients with endometrial carcinoma. For this purpose, all tumours were stained immunohistochemically with oestrogen receptor and Ki-67 monoclonal antibodies. Oestrogen receptor expression and proliferation indices were found to be statistically associated with grade, age, menopausal status, vascular invasion and tumour size ( p < 0.001). Quantitative assessment of tumour proliferation and expression of oestrogen receptor were found to be important prognostic indicators in endometrial adenocarcinoma. PMID- 17130036 TI - Vaginal tape erosion following transobturator tape (TOT) operation for stress urinary incontinence. AB - Inorganic sling materials when used for treatment of urinary stress incontinence have a high success rate of achieving continence. They unfortunately attract the complication of vaginal tape erosion. When used for Transobturator tape procedure (TOT) we found the Obtape sling (Mentor-Porges, Le Plessis-Robinson, France) attracted a high rate of erosion, likely due to the tape properties. We describe a series of tape erosions with the Obtape sling, the presentation, conservative and surgical management with favourable rates of maintenance of continence. We suggest the use of larger pore sized tapes to decrease the occurrence of vaginal tape erosion. PMID- 17130037 TI - Thermal balloon endometrial ablation (Cavaterm) in the management of menorrhagia. PMID- 17130038 TI - Spontaneous ovarian bleeding in patients with congenital coagulopathy: case series and review of the literature. PMID- 17130039 TI - Recurrent variegate porphyria in a pregnant woman. PMID- 17130040 TI - Dissociative disorder during pregnancy. PMID- 17130041 TI - Abducens nerve palsy complicating pregnancy. PMID- 17130042 TI - Pregnancy and galactosaemia. PMID- 17130043 TI - Breus' mole in pregnancy. PMID- 17130044 TI - Alcoholic pancreatitis in pregnancy. PMID- 17130045 TI - Acute vulval oedema with urinary retention in pregnancy. PMID- 17130046 TI - Partial molar pregnancy with severe pre-eclampsia at 19 weeks' gestation. PMID- 17130047 TI - Fetal akinesia deformation sequence. Pena-Shokeir type I syndrome: new features of an un-uncommon condition. PMID- 17130048 TI - Intrauterine pregnancy after microwave endometrial ablation. PMID- 17130049 TI - Extensive bilateral iliofemoral and inferior vena cava thrombosis following caesarean section. PMID- 17130050 TI - Subcutaneous emphysema and pneumomediastinum in labour. PMID- 17130051 TI - Rectus sheath haematoma in pregnancy: a clinical challenge. PMID- 17130052 TI - Prolapsed cervical fibroid in pregnancy: a challenging obstetric dilemma. PMID- 17130053 TI - Toxic shock syndrome: a silent killer. PMID- 17130054 TI - Vibrator in the peritoneal cavity: a case of post-hysterectomy vaginal vault evisceration. PMID- 17130055 TI - Uterine rupture following a second trimester medical termination of pregnancy in a woman with a previous caesarean section. PMID- 17130056 TI - How innocent is the vaginal pessary? Two cases of vaginal cancer associated with pessary use. PMID- 17130057 TI - Secondary amenorrhoea associated with an isolated elevated serum leuteinising hormone: an unusual presentation of a granulosa cell tumour. PMID- 17130058 TI - Endocervical stromal sarcoma. PMID- 17130059 TI - Haematometra following large loop excision of the transformation zone. PMID- 17130060 TI - Familial cystic hygroma with normal karyotype. PMID- 17130061 TI - Re: Ghaem-Maghami S, Brockbank E, Bridges J. 2006. Survey of surgical experience during training in obstetrics and gynaecology in the UK. Journal of Obstetrics and Gynaecology 26(4):297 - 301. PMID- 17130063 TI - Understanding and tackling health-related stigma. PMID- 17130064 TI - International research workshop on health-related stigma and discrimination. PMID- 17130065 TI - Health-related stigma: rethinking concepts and interventions. AB - As a feature of many chronic health problems, stigma contributes to a hidden burden of illness. Health-related stigma is typically characterized by social disqualification of individuals and populations who are identified with particular health problems. Another aspect is characterized by social disqualification targeting other features of a person's identity-such as ethnicity, sexual preferences or socio-economic status-which through limited access to services and other social disadvantages result in adverse effects on health. Health professionals therefore have substantial interests in recognizing and mitigating the impact of stigma as both a feature and a cause of many health problems. Rendering historical concepts of stigma as a discrediting physical attribute obsolete, two generations of Goffman-inspired sociological studies have redefined stigma as a socially discrediting situation of individuals. Based on that formulation and to specify health research interests, a working definition of health-related stigma is proposed. It emphasizes the particular features of target health problems and the role of particular social, cultural and economic settings in developing countries. As a practical matter, it relates to various strategies for intervention, which may focus on controlling or treating target health problems with informed health and social policies, countering the disposition of perpetrators to stigmatize, and supporting those who are stigmatized to limit their vulnerability and strengthen their resilience. Our suggestions for health studies of stigma highlight needs for disease- and culture specific research that serves the interests of international health. PMID- 17130066 TI - Sociology, social structure and health-related stigma. AB - There is a long and cross-disciplinary tradition of analysing chronic and disabling illness in terms of relations of stigma. The present paper offers a sociological approach which emphasizes: (a) the causal importance of social structures for grasping stigma relations; (b) the importance of understanding stigma relations in the context of wider societal change; and (c) the ways in which relations of stigma typically interact with other relations, such as those of class and command. It is suggested that consideration of specific and often condition-related strategies to reduce stigma might profitably be set in such a context. PMID- 17130067 TI - A psychological model of social control and stigmatization: evolutionary background and practical implications. AB - The strengths and limitations of three main theoretical approaches to social control and stigmatization are discussed, originating from social psychology, sociology and anthropology, and evolutionary psychology, respectively. A theoretical model is proposed that integrates universal (evolutionary derived) psychological aspects of social control and the great variation that can be observed in responding to deviance within and between cultures and historical periods. Practical implications of the model are discussed. PMID- 17130068 TI - Measuring health-related stigma--a literature review. AB - Stigma related to chronic health conditions such as HIV/AIDS, leprosy, tuberculosis, mental illness and epilepsy is a global phenomenon with a severe impact on individuals and their families, and on the effectiveness of public health programmes. To compare stigma measurement in different disciplines, a literature review was conducted. References were obtained through a search of literature databases and through examining relevant bibliographies. Sixty-three papers were selected that addressed the issue of measurement of stigma or related constructs and that contained a sample of the instrument or items used. Five unpublished studies were also included in the review. The aspects of health related stigma used for assessment can be grouped in five categories. First, the experience of actual discrimination and/or participation restrictions on the part of the person affected; second, attitudes towards the people affected; third, perceived or felt stigma; fourth, self or internalized stigma; and fifth, discriminatory and stigmatizing practices in (health) services, legislation, media and educational materials. Within each of these areas, different research methods have been used, including questionnaires, qualitative methods, indicators and scales. The characteristics of the instruments considered most promising are described and compared. The purpose of stigma assessment is to increase our understanding of stigma and its determinants and dynamics, to determine its extent or severity in a given setting or target group and to monitor changes in stigma over time. The conclusions from this review are that (a) the consequences of stigma are remarkably similar in different health conditions, cultures and public health programmes; (b) many instruments have been developed to assess the intensity and qualities of stigma, but often these have been condition-specific; and (c) development of generic instruments to assess health-related stigma may be possible. To achieve this aim, existing instruments should be further validated, developed or adapted for generic use, where possible. PMID- 17130069 TI - Measuring HIV stigma: existing knowledge and gaps. AB - The growing recognition of the reduction of HIV stigma as central to effective programs across the HIV/AIDS prevention to care and treatment continuum is leading to an increasing number of programs focused on stigma reduction. Correctly evaluating the impact of these programs depends on having a good set of measures that effectively capture and distinguish the complexities of HIV stigma. This paper reviews the existing literature on HIV stigma measurement and identifies key gaps that remain. There is a need for measures at the general population level that are unambiguous about the cause of the stigmatizing behavior, that capture enacted stigma (discrimination), and that can distinguish compound (layered) stigma. In addition, studies are needed in a wider variety of contexts and on a larger scale that include a comprehensive set of measures to capture the complexity of HIV-related stigma and ensure appropriate evaluation of stigma-reduction programs. PMID- 17130070 TI - Assessing the stigma of tuberculosis. AB - Tuberculosis (TB) stigma is considered a vague concept, because it is not easily assessed. The purpose of this article is to review methodological approaches for assessing TB stigma. We reviewed the published studies and the gray literature. We also present the main features of a qualitative and quantitative assessment of TB stigma, which has been done in one specific project in Nicaragua. As a whole, we illustrate the variety of TB stigma in terms of domains, consequences, determinants and methods used; some features emerge more frequently than others from the studies. However, results of the review show a relative scarcity of the TB stigma assessment experiences, mainly in looking at the consequences for the TB patient and in the use of quantitative methods and scales. Additional assessment studies in diverse contexts are needed so that stigma will be considered a priority in the organization of care for people affected by tuberculosis. PMID- 17130071 TI - The fight against stigma: an overview of stigma-reduction strategies and interventions. AB - In many health conditions, people are severely affected by health-related stigma and discrimination. A literature review was conducted to identify stigma reduction strategies and interventions in the field of HIV/AIDS, mental illness, leprosy, TB and epilepsy. The review identified several levels at which interventions and strategies are being implemented. These are the intrapersonal, interpersonal, organizational/institutional, community and governmental/structural level. Although a lot of work has been carried out on stigma and stigma reduction, far less work has been done on assessing the effectiveness of stigma-reduction strategies. The effective strategies identified mainly concentrated on the individual and the community level. In order to reduce health-related stigma and discrimination significantly, single-level and single target group approaches are not enough. What is required is a patient-centred approach, which starts with interventions targeting the intrapersonal level, to empower affected persons to assist in the development and implementation of stigma-reduction programmes at other levels. PMID- 17130072 TI - Interventions to reduce epilepsy-associated stigma. AB - Epilepsy-associated stigma has long been recognized as a significant cause of psychosocial morbidity for people with epilepsy. This paper reviews the available literature addressing interventions aimed at reducing epilepsy-associated stigma and briefly discusses future that may assist in the development of effective interventions. PMID- 17130073 TI - Interventions to address the stigma associated with leprosy: a perspective on the issues. AB - This paper presents a perspective on stigma as an effect of leprosy. It identifies some of the strengths and weaknesses of current approaches to stigma reduction in leprosy and presents a rationale for considering alternative strategies. It is suggested that models used to explain health behavior in developed societies are inappropriate for explaining leprosy stigma or for developing strategies to address it. The author recommends due consideration of the alternative logic that characterizes cultural belief systems in countries where leprosy is a challenge. Criticism of the common practice of information dissemination as a strategy to address leprosy stigma is defended and the merits and limitations of an integrated health service in India is discussed. The author defends the suggestion that the principal objective of stigma interventions should be "normalization". An example of a Nepalese project based on empowerment theory is given to demonstrate how the transformation of identity from outcast to positive change agent, can effect "normalization". PMID- 17130074 TI - Social interventions to moderate discriminatory attitudes: the case of the physically challenged in India. AB - Disability attitudes are major barriers in improving life conditions of physically challenged people in developing countries. Studies have shown that people, in general, harbour negative and paternalistic attitudes towards persons with disabilities. Myths, legends, scriptures and folklores are all part of the cultural belief system that shape such attitudes. The physically challenged frequently suffer more due to societal prejudices than due to their disabling physical conditions. The social and physical environment in which these physically challenged live is often designed without much consideration of their special needs. In this article, some social interventions aimed at changing disability attitudes of rural people in India are discussed. As part of a community-based rehabilitation programme, the main thrust of these interventions was to shift attention of local communities from disabilities to abilities of the physically challenged. The efficacy of these social interventions is discussed. PMID- 17130075 TI - Lessons from a 10-year global programme against stigma and discrimination because of an illness. AB - The World Psychiatric Association has been carrying out a major international programme aiming to reduce stigma and discrimination because of schizophrenia. The programme is under way in 19 countries. The paper gives a brief description of the programme and lists the lessons learned during its course. PMID- 17130076 TI - A young woman's distress. PMID- 17130077 TI - On Nachtraglichkeit: the modernity of an old concept. AB - Nachtraglichkeit provides the memory, not the event, with traumatic significance and signifies a circular complementarity of both directions of time. Conceived by Freud as early as 1895 in the Project for a scientific psychology, the concept remains in his work without official status but through its character of biphasic development and latency indispensable for understanding temporal connections and psychic causality. As an implicit principle it is linked with the postponement and biphasic onset of sexual life and retains its sometimes hidden importance until the late Moses study. Temporarily virtually forgotten, it was recalled to memory by Lacan in 1953. Translations into French as apres-coup and into English as 'deferred action' emphasized the two vectors (retroactivity and after-effect) separately which are united in the substantive form coined by Freud. Unnoticed, it played a part in many aspects of clinical practice, especially in Winnicott's Fear of breakdown and the subsequent (nachtraglich) working through of unconscious infantile and transgenerational conflicts. The author uses a clinical illustration to elucidate the belated understanding of the striving for non existence in Winnicott's sense. Wolfgang Loch extended Freud's concept of Nachtraglichkeit in a constructivist way, advocating an art of interpretation as an innovative enterprise through which connections are not only unmasked but also created, constituted by subsequent (nachtraglich) reinterpretation of a subjective past. Very briefly the author refers to the interdisciplinary reception of the concept of Nachtraglichkeit, especially in cultural studies. PMID- 17130078 TI - A shadow concept. AB - The author focuses on the significance of preconscious thinking, and its relationship to what we think of as unconscious fantasies. He reopens Freud's forgotten struggle with preconscious thinking, while he explores preconscious thinking as the basis for thinking about psychoanalytic treatment. This includes our goals in bringing an idea to the analysand's attention, and the role of transitional space where thoughts and feelings can be played with. PMID- 17130079 TI - After pluralism: towards a new, integrated psychoanalytic paradigm. AB - After a restatement of the isolationism of psychoanalysis from allied disciplines, and an examination of some of the reasons for the diversity of schools of thought and the fragmentation of psychoanalytic knowledge, the author suggests the need to adopt principles of correspondence or external coherence along with those of hermeneutic coherence to validate psychoanalytic hypotheses. Recent developments in neurocognitive science have come to the aid of psychoanalysis in this period of crisis, resulting in the proposition of integrating both areas to form a new paradigm for the construction of the theory of the mind. This emerging paradigm tries to integrate clinical knowledge with neurocognitive science, findings from studies on the process and outcome of psychotherapy, research into the early mother-infant relationship, and developmental psychopathology. The author examines theoretical- technical models based on the concept of drives and of relationships in the light of interdisciplinary findings. He concludes that the relational model has a broad empirical base, except when the concept of drives is discredited. Interdisciplinary findings have led to the positing of the replacement of the Freudian model of drives with a model of motivational systems centred on affective processes. He draws certain conclusions which have a bearing on the technique of psychoanalytic treatment. These arise from the adoption of the new integrated paradigm. PMID- 17130080 TI - Dream interpretation, affect, and the theory of neuronal group selection: Freud, Winnicott, Bion, and Modell. AB - The author uses a dream specimen as interpreted during psychoanalysis to illustrate Modell's hypothesis that Edelman's theory of neuronal group selection (TNGS) may provide a valuable neurobiological model for Freud's dynamic unconscious, imaginative processes in the mind, the retranscription of memory in psychoanalysis, and intersubjective processes in the analytic relationship. He draws parallels between the interpretation of the dream material with keen attention to affect-laden meanings in the evolving analytic relationship in the domain of psychoanalysis and the principles of Edelman's TNGS in the domain of neurobiology. The author notes how this correlation may underscore the importance of dream interpretation in psychoanalysis. He also suggests areas for further investigation in both realms based on study of their interplay. PMID- 17130081 TI - Attachment theory and psychoanalysis: some remarks from an epistemological and from a Freudian viewpoint. AB - The author examines Bowlby's attachment theory and more recent versions of it from an epistemological viewpoint and subjects it to questioning on whether they are in line with central concepts of Freudian psychoanalysis. He argues that Bowlby's basic tenets regarding attachment theory, which later attachment theorists never seriously questioned, do not conform to scientific standards, and that psychoanalytic issues such as the dynamic unconscious, internal conflicts, interaction of drive wishes and the role of defence in establishing substitutive formations are either ignored or not treated in sufficient depth. In the light of this, Fonagy's assertion that psychoanalytic criticism of attachment theory arose from mutual misunderstandings and ought nowadays to be seen as outdated is reversed: psychoanalytic criticism can only be regarded as outdated if either basic tenets of Freudian psychoanalysis, or attachment theory or both are misunderstood. PMID- 17130082 TI - A servant's bargain: perversion as survival. AB - The author views psychological perversion as a ubiquitous adaptation which preserves the person's capacity to apprehend reality while changing its meaning at will, in the service of staving off unbearable emotional pain. This is understood as derivative of the infant's horror of the nameless dread uncontained by the tantalizing environment. Whether perceived or actual, parental abdication of their protective responsibility (the meaning of the generational divide) can threaten the child's capacity to learn to contain his affects, and can bring forth the creation of a fetishistic artificial container as a substitute for a reliable connection. In this way, fantasied instant unison might take the place of genuine intersubjectivity. Using clinical examples from the analysis of Mr. A, and Bion's concept of reversible perspective, the author depicts the challenge presented by the patient's rigid and invisible commitment to turn the possibility of an interpersonal connection into yet another incarnation of the familiar script, in which decency and good will are silently and reflexively deformed. In this approach, perversion is seen not simply as the cynical game it frequently becomes, but as a person's last effort to protect himself from anticipated psychological breakdown, in which excitement serves as a smokescreen hiding the internal terror. The technical difficulties arise from the paradox of the patient's adamant refusal of a healthier relationship, which can be useful to see as an imperative form of survival, rather than merely an intrinsic malignancy. PMID- 17130083 TI - Julie's museum: the evolution of thinking, dreaming and historicization in the treatment of traumatized patients. AB - The author contends that, following Freud, trauma may be viewed as a disruption of the ego's 'protective shield' and that a central factor of this shield is an internalized relationship to a thinking-containing mother. Severe trauma destroy this inner connection, resulting in the reversal of a-function and the establishment of a rigid traumatic organization (ss-screen) that brings coherence to the shattered psyche. However, this is an 'organized chaos' in which concrete forms of thinking predominate. The patient's ability to think, dream and imagine is significantly curtailed and she is consequently locked in a traumatic world from which she is unable to evolve. He offers a detailed case history to illustrate these points and the vital role the analyst's imaginative capacities play in the analysis of such individuals. Finally, he addresses the development of the capacity to represent the trauma, starting with primitive, often somatically encoded experiences, and evolving toward the capacity for historicization. PMID- 17130084 TI - All holes are the same: emerging from the confusion. AB - The author describes a type of mental development in which the primitive libidinal value of faeces and urine is kept unaltered all through adulthood. In this instance, individuals harbour fantasies of projective identification with the internal parents, denying their real dependence, which leads to a pseudomaturity. These fantasies are usually accompanied by anal and genital masturbation and have intense aggressive content. Furthermore, there is a significant confusion among all body orifices, leading to an inability to distinguish the self from the object, the inner world from outside reality. Individuals with this type of development can only maintain limited object relations and have highly erotized excretory activities. Manipulation of faeces may occur. This structure works as a defence against breakdown, but may also provide pleasure as it gives the illusion of omnipotent control of the object, as typical in pathological organizations. The author presents clinical material and discusses the difficulties of the analysis, in which the patient tries to seduce the analyst into colluding with his mental functioning, primarily as a means of communication as well as a projection of his despair and his disbelief in the analyst's ability to help him. PMID- 17130085 TI - Where are you, my beloved? On absence, loss, and the enigma of telepathic dreams. AB - The subject of dream telepathy (especially patients' telepathic dreams) and related phenomena in the psychoanalytic context has been a controversial, disturbing 'foreign body' ever since it was introduced into psychoanalysis by Freud in 1921. Telepathy- suffering (or intense feeling) at a distance (Greek: pathos + tele)-is the transfer or communication of thoughts, impressions and information over distance between two people without the normal operation of the recognized sense organs. The author offers a comprehensive historical review of the psychoanalytic literature on this controversial issue, beginning with Freud's years-long struggles over the possibility of thoughttransference and dream telepathy. She then describes her own analytic encounter over the years with five patients' telepathic dreams-dreams involving precise details of the time, place, sensory impressions, and experiential states that the analyst was in at that time, which the patients could not have known through ordinary sensory perception and communication. The author's ensuing explanation combines contributory factors involving patient, archaic communication and analyst. Each of these patients, in early childhood, had a mother who was emotionally absent-within-absence, due to the absence of a significant figure in her own life. This primary traumatic loss was imprinted in their nascent selves and inchoate relating to others, with a fixation on a nonverbal, archaic mode of communication. The patient's telepathic dream is formed as a search engine when the analyst is suddenly emotionally absent, in order to find the analyst and thus halt the process of abandonment and prevent collapse into the despair of the early traumatization. Hence, the telepathic dream embodies an enigmatic 'impossible' extreme of patient-analyst deep-level interconnectedness and unconscious communication in the analytic process. This paper is part of the author's endeavour to grasp the true experiential scope and therapeutic significance of this dimension of fundamental patient-analyst interconnectedness. PMID- 17130086 TI - 'The newspaper reader': on the meaning of concrete objects in a psychoanalytic treatment. AB - The author investigates the meaning of concrete objects in the psychoanalytic treatment of a severely disturbed patient for the development of his inner world and the analytic process. She includes a survey of relevant theoretical concepts with an emphasis on Winnicott and Bion. It is shown that the objects served basic defensive functions both within the analytic relationship and for the precarious intrapsychic state of the patient. The author describes the technical dealing that led to a structural change. From the comparison of the initial dream and a later dream, Mr N's inner development from total inclusion in the object to triadic reality of separated, repaired objects becomes discernible. The author shows how this progress was facilitated by his use of concrete objects as links between his psychotic and non-psychotic parts, as well as by the specific way the analyst handled the paradoxical transference- countertransference. She also illustrates the thesis that the developmental steps described are crucial for the capability to digest psychic pain by symbolization instead of discharging it in a destructive-violent way. PMID- 17130087 TI - The coming changes in psychoanalytic education: part I. AB - The author presents a critical overview of the literature on psychoanalytic education, focusing on criticism regarding structural aspects of our educational institutions. He then presents arguments for the need of radical changes in the organizational structure of institutes, and focuses on the problems of the training analysis system. He proposes concrete solutions for these problems, in the form of changes both in the assignment of responsibilities for the personal analysis of candidates and in the selection and function of supervisors. PMID- 17130088 TI - Authority as paradox: the transformations of Don Quijote. AB - The author's contention is that the analysand's temporary attribution of authority to the analyst is inherent in the analytic situation; this is seen as a transitional and paradoxical form of authority pertaining neither to internal nor external reality, but dwelling in the analytic third. The author proposes a conceptualization of psychoanalytic authority as a form of aesthetic authority according to Gadamer's definitions. While the scientific and hermeneutic codes for the understanding of authority in psychoanalysis assume that the main issue at stake is the delimitation of the objectivity or the subjectivity of the analyst's knowledge, this aesthetic perspective centres on the analysand's attribution of a claim of truth to analytic interpretations, and on the experience of recognition. The experience of recognition of a possible truth is particular and context-bound, as well as self-transformational. A reading of three episodes from Cervantes's The history of Don Quixote de la Mancha illuminates the transitional and paradoxical character of aesthetic authority within a transformational dialogue. These episodes are read as dramatizations of different positions vis-a-vis the paradoxical authority that characterizes transformational dialogues. PMID- 17130089 TI - The use of the past and the present in the clinical setting: pasts and presents. AB - The author provides a brief summary of Latin American literature concerning temporality. She shows that a common theme throughout all these papers is that the analytical relationship is considered to be bipersonal and symmetrical, thus demonstrating a concern for establishing the basis by which social subjectivity can be reconsidered. This literature displaces the idea of linear time from its central position and introduces other measures of time. The analytical relationship takes place not only in the past but also in a newly created present. This is the ongoing present, of what is happening now, instantaneous and without a prior history attached to it. This leads the author to suggest that there is a present to one's history and a history to one's present. She then analyses the consequences of this proposition by examining some clinical material where she attempts to pinpoint those instances in which the analyst may have reacted defensively, tending to position himself in the analysand's past instead of being able to take action in the present. Clinical material from the IJP is used. PMID- 17130090 TI - Identity, destiny and terrorism: the effect of social terror on identity formation. PMID- 17130091 TI - Resistance as a response to trauma in the clinical moment: the approaches of a London Kleinian and American ego-psychologist. PMID- 17130092 TI - On: reading Loewald: Oedipus reconceived. PMID- 17130095 TI - Bicalutamide 150 mg in addition to standard care for patients with early non metastatic prostate cancer: updated results from the Scandinavian Prostate Cancer Period Group-6 Study after a median follow-up period of 7.1 years. AB - OBJECTIVE: The Early Prostate Cancer (EPC) programme is evaluating the efficacy and tolerability of bicalutamide following standard care (radiotherapy, radical prostatectomy or watchful waiting) in patients with localized (T1-2, N0/Nx) or locally advanced (T3-4, any N; or any T, N + ) non-metastatic prostate cancer. Herein we report the latest findings after a median follow-up period of 7.1 years from the Scandinavian Prostate Cancer Group (SPCG)-6 study, one of three trials in the EPC programme. MATERIAL AND METHODS: A total of 1218 patients were randomized on a 1:1 basis to either bicalutamide 150 mg/day (n=607) or placebo (n=611) following standard care; 81.4% were followed conservatively (watchful waiting). The primary endpoints were objective progression-free survival (PFS) and overall survival (OS). RESULTS: In patients with localized disease there was no significant difference in PFS [hazard ratio (HR) 0.85; 95% CI 0.69-1.06; p=0.15] and a trend towards decreased OS with bicalutamide plus standard care compared with standard care alone (HR 1.23; 95% CI 0.96-1.58; p=0.11). In patients with locally advanced disease, bicalutamide significantly improved PFS, reducing the risk of progression by 53% compared with standard care alone (HR 0.47; 95% CI 0.37-0.59; p<0.001). The median time to progression was 8.8 years for bicalutamide plus standard care and 7.1 years for standard care alone. There was a significant improvement in OS with bicalutamide plus standard care, with a reduction in the risk of death of 35% versus standard care alone (HR 0.65; 95% CI 0.50-0.85; p=0.001). CONCLUSION: This analysis of the SPCG-6 study showed that bicalutamide plus standard care offers significant PFS and OS benefits for patients with locally advanced disease, but not for those with localized disease. PMID- 17130096 TI - Karolinska prostatectomy: a robot-assisted laparoscopic radical prostatectomy technique. AB - OBJECTIVE: The last decade has witnessed an increasing trend towards minimally invasive management of prostate cancer, including laparoscopic and, more recently, robot-assisted laparoscopic prostatectomy. Several different laparoscopic approaches have been continuously developed during the last 5 years and it is still unclear which technique yields the best outcome. We present our current technique of robot-assisted laparoscopic radical prostatectomy. MATERIAL AND METHODS: The technique described has evolved during the course of >400 robotic prostatectomies performed by the robotic team since the robot-assisted laparoscopic radical prostatectomy program was introduced at Karolinska University Hospital in January 2002. SURGICAL PROCEDURE: Our procedure comprises several modifications of previously reported ones, and we utilize fewer robotic instruments to reduce costs. An extended posterior dissection is performed to aid in the bladder neck-sparing dissection. In nerve-sparing procedures the vesicles are divided to avoid damage to the erectile nerves. In order to preserve the apical anatomy the dorsal venous complex is incised sharply and is first over sewn after the apical dissection is completed. CONCLUSIONS: Our technique enables a more fluent dissection than previously described robotic techniques. Minimizing changes of instruments and the camera not only cuts costs but also reduces inefficient operating maneuvers, such as switching between 30 degrees and 0 degrees lenses during the procedure. We present a technique which in our hands has achieved excellent functional and oncological results. PMID- 17130097 TI - Prediction of extracapsular extension of prostate cancer based on systematic core biopsies. AB - OBJECTIVE: To evaluate the clinical utility of transrectal ultrasound-guided systematic sextant or octant biopsies for the prediction of extracapsular extension (ECE) at radical prostatectomy. MATERIAL AND METHODS: We performed a retrospective analysis of 84 patients who underwent preoperative staging and transrectal ultrasound-guided systematic sextant (n=60) or octant (n=24) biopsy. The presence of ECE was correlated with the number of positive biopsies on each side of the prostate by chi(2) analysis. Sensitivity, specificity and positive and negative predictive values were calculated for both positive (two or three positive biopsies per side) and negative (none or one positive biopsy per side) test results. The number of positive cores was thereafter combined with two other parameters: prostate-specific antigen (PSA) level and Gleason score. RESULTS: ECE was evidenced at radical prostatectomy in 24% of patients (20/84). chi(2) analysis demonstrated a significant correlation between the number of positive biopsies and the presence of ECE. Analysis of the 168 prostate sides and dominant sides revealed that systematic needle biopsies had positive predictive values of 46.7% and 37%, respectively and negative predictive values of 89% and 94%, respectively. Use of a combination of parameters (biopsy Gleason score > or =7 vs <7; PSA >10 vs < or = 10 ng/ml; and >1 positive core vs none or one positive cores) identified patients at high or low risk of ECE. At the extremes, none of the 10 patients in the low-risk group had ECE at radical prostatectomy, compared to 77% of those in the high-risk group. CONCLUSION: The probability of ECE at radical prostatectomy can be accurately predicted based on the number of positive sextant and octant biopsies, either alone or in combination with other parameters. PMID- 17130098 TI - Prediction of percent Gleason grade 4/5 by multiple core biopsies. AB - OBJECTIVE: To evaluate whether percent Gleason grade 4/5 (i.e. the proportion of a tumor occupied by high-grade cancer) can be predicted by multiple needle biopsies. MATERIAL AND METHODS: In 115 men, 8-14 (mean 10) biopsies were taken, including eight from standardized positions (apex, mid-medial, mid-lateral and base). Biopsies were reviewed and cancer lengths measured. All men underwent radical prostatectomy. The prostatectomy specimens were totally embedded and tumor volume measured planimetrically. Gleason scores and percent Gleason grade 4/5 were assessed for both biopsy and prostatectomy specimens. RESULTS: Percent Gleason grade 4/5 in prostatectomy specimens was predicted correctly in 34% of cases and within 10%, 20% and 30% in 55%, 64% and 73% of cases, respectively. Biopsies had a sensitivity, specificity and accuracy for Gleason grade 4/5 of 62%, 87% and 69%, respectively. Positive and negative predictive values were 93% and 45%, respectively. The weighted kappa value for agreement was slightly higher for Gleason score (0.685) than for percent Gleason grade 4/5 (0.573). The univariate correlation for percent Gleason grade 4/5 in biopsies and the main tumor was r=0.62, r(2)=0.39 (p<0.001). In univariate logistic regression, percent Gleason grade 4/5 on biopsies predicted the presence of any Gleason grade 4/5 cancer in the main tumor (p=0.009). CONCLUSIONS: Gleason grade 4/5 in prostatectomy specimens correlates with findings in preoperative biopsies. Whether this measure will be used in routine practice remains to be seen. PMID- 17130099 TI - Afferent intussuscepted antireflux nipple valve complications in the Kock pouch for continent urinary diversion Early results with a modified technique. AB - OBJECTIVES: To analyse the risk of afferent nipple valve stenosis and its consequences and management in patients with a Kock pouch for continent urinary diversion and to study the early results after using an alternative antireflux technique. MATERIAL AND METHODS: Sixty patients consecutively operated on with a conventional Kock reservoir for continent cutaneous urinary diversion between 1988 and 2001 were analysed with regard to the occurrence of afferent nipple valve stenosis and its clinical characteristics. Sixteen patients operated on for continent urinary diversion during the period 2002-04 had the antireflux valve constructed according to the serous-lined extramural ileal valve technique. RESULTS: Eight patients with a conventional Kock pouch developed true afferent nipple valve stenosis and the risk approached 30% after 15 years. Dilatation and stenting were usually successful. CONCLUSIONS: The high risk of afferent nipple valve stenosis when using the intussuscepted nipple valve in the construction of a Kock reservoir for continent cutaneous urinary diversion calls for an alternative method for anastomosing the ureters to the reservoir. Our early results with the combined Kock/T-pouch are promising. PMID- 17130100 TI - Impact of diagnostic and treatment delay on survival in patients with renal pelvic and ureteral cancer. AB - OBJECTIVE: To investigate the relationships between diagnostic and treatment delay and tumour stage and survival among patients with malignant tumours in the renal pelvis and ureter. MATERIAL AND METHODS: A clinical and histopathological review was performed on 943 patients with a primary malignant tumour in the renal pelvis and ureter. We selected 394 patients who had macrohaematuria as an initial symptom, had no previous history of bladder cancer, had undergone surgery and had adequate follow-up. We performed uni- and multivariate analyses of prognostic factors for disease-specific survival. RESULTS: The median number of days between the first occurrence of macrohaematuria and surgery was 83.5 days (range 4-1770 days). Patients with advanced tumours had the shortest median delay. Advanced tumour stage, a solid growth pattern and vascular invasion were of prognostic importance for disease-specific survival in the multivariate analysis, but diagnostic and treatment delay were not. CONCLUSIONS: Although the delay was unacceptably long it still had no impact on survival, probably because macroscopic haematuria is a late symptom, in particular in high-grade tumours. New screening methods for the early detection of cancer and new treatment modalities are needed to improve the poor prognosis in stage pT3-pT4 tumours. PMID- 17130101 TI - Impact on quality of life of different lower urinary tract symptoms in men measured by means of the SF 36 questionnaire. AB - OBJECTIVE: To describe how different lower urinary tract symptoms (LUTS) affect the quality of life (QOL) in men. MATERIAL AND METHODS: The study included 1008 men aged 40-80 years living in the community of Surahammar, Sweden who had answered a questionnaire concerning stress incontinence, urgency and post micturition dribbling 12 months earlier. The occurrence and severity/frequency of 12 specific LUTS were assessed using the Danish Prostatic Symptom Score questionnaire. QOL was evaluated using the Short Form 36 (SF-36) questionnaire. RESULTS: Post-micturition dribbling was the most frequently reported symptom (71%) and stress incontinence the least common (11%). Men who experienced urge, stress or "other incontinence" had lower mean scores for all of the eight dimensions measured by the SF-36 than men without such symptoms. Furthermore, men who experienced a moderate/severe degree of weak stream or nocturia reported a poorer QOL for all dimensions compared to men with a mild level of the same symptoms. QOL was found to decrease with increasing age. Men aged 66-80 years with "other incontinence" reported lower mean SF-36 scores for physical functioning, role physical, role emotional, social functioning and body pain than 40-65-year-old men. CONCLUSIONS: LUTS in men affect QOL dimensions differently. Storage symptoms appear to reduce QOL more than voiding and post-micturition symptoms. Urinary incontinence affected all eight of the dimensions evaluated. Elderly men with LUTS reported a lower QOL than younger men. PMID- 17130102 TI - Under what conditions is feedback microwave thermotherapy (ProstaLund Feedback Treatment) cost-effective in comparison with alpha-blockade in the treatment of benign prostatic hyperplasia and lower urinary tract symptoms? AB - OBJECTIVE: Lower urinary tract symptoms (LUTS) due to benign prostatic hyperplasia (BPH) are a common condition in men, and their incidence increases with age. The objective of this study was to evaluate the potential cost-utility of microwave thermotherapy, specifically the ProstaLund Feedback Treatment (PLFT), versus alpha-blockade in Swedish patients with LUTS due to BPH. MATERIAL AND METHODS: A health-economic simulation model, based on long-term disease progression and costs, was developed to analyse the cost-utility of PLFT in comparison with alpha-blockade over a 3-year period based on data from published literature, treatment programmes and official price lists. Outcome measures used in the analysis were quality of life, survival and reduction in International Prostate Symptom Score. Sensitivity analyses were performed for a number of essential variables. The perspective of the study is the healthcare sector. All costs are expressed as 2003 prices. RESULTS: Three years after an intervention with PLFT or initiation of drug treatment the cost-utility of PLFT was estimated at approximately euro 6600-9500 per quality-adjusted life-year gained. The cost utility was further improved over a longer time period, and PLFT appears to be cost-saving after 5 years. One important finding from the model simulation was that PLFT also seems to be favourable in patients with less pronounced symptoms. This result may be further validated when additional results from controlled clinical trials become available. CONCLUSIONS: The present model simulation indicates that treatment with PLFT seems to be cost-effective compared with drug therapy with alpha-blockade. The result shows that the time-frame of the analysis has a great impact on the cost-effectiveness ratio. PMID- 17130103 TI - Relationship between nocturnal penile tumescence parameters, International Index of Erectile Function symptom scores and sildenafil responses1This paper was presented at the 6th Congress of the International Society of Sexual and Impotence Research, Buenos Aires, Argentina, 17-21 October 2004. AB - OBJECTIVE: To evaluate the correlation between International Index of Erectile Function (IIEF) questionnaire scores, nocturnal penile tumescence parameters and sildenafil response in patients with erectile dysfunction using a minimal investigations approach. MATERIAL AND METHODS: A total of 97 patients without any risk factors for erectile dysfunction were included. After completing the IIEF questionnaire, nocturnal penile tumescence monitoring was performed in all patients for 1 or 2 nights. Subsequently, sildenafil was given at a dosage of 50 or 100 mg according to the response. The relationship between nocturnal penile tumescence results, the severity of erectile dysfunction according to the IIEF questionnaire and sildenafil response was evaluated using Pearson's chi2 test. RESULTS: Based on the questionnaire scores, 25 patients (25.8%) had mild, 44 (45.4%) had moderate and 28 (28.8%) had severe erectile dysfunction. Forty-four patients (45.4%) had normal and 53 (54.6%) had abnormal parameters after nocturnal penile tumescence recording. The sildenafil response was positive in 63 patients (64.9%) who were given 50 mg and in 26 (26.8%) who were given 100 mg. However, eight patients (8.2%) did not respond to sildenafil. While the severity of erectile dysfunction according to IIEF scores and the sildenafil response did not show any correlations with nocturnal penile tumescence results, there was a statistically significant relationship between sildenafil response and the severity of erectile dysfunction. CONCLUSION: Changing concepts in the evaluation of ED have resulted in the use of effective therapeutic applications consistent with the degree of symptoms of patients without the waste of time caused by performing expensive, invasive, ineffective and time-consuming diagnostic tests. PMID- 17130105 TI - Urinary excretion of metabolizable acid in response to non-metabolizable acid ingestion. PMID- 17130106 TI - Accuracy of uroflowmetric parameters and post-void residual volume. PMID- 17130107 TI - Resolution of proteinuria in a patient with X-linked Alport syndrome treated with cyclosporine. AB - We report a 9-year-old Icelandic male with Alport syndrome and nephrotic-range proteinuria who responded well to cyclosporine therapy. He presented at the age of 2 years with gross hematuria and proteinuria during an episode of upper respiratory tract infection. Three years later he had developed persistent proteinuria; kidney function was normal. A renal biopsy revealed marked irregularities in the glomerular basement membrane consistent with Alport syndrome. Mutation analysis revealed a single base insertion in COL4A5 which was predicted to cause a major structural defect in the collagen IV alpha5 chain. Despite angiotensin-converting enzyme inhibitor therapy his proteinuria progressed to the nephrotic range associated with edema. At the age of 7 years, cyclosporine therapy was instituted, which promptly resulted in almost complete resolution of proteinuria. Three years later his urinary protein excretion was close to the normal range and serum creatinine remained within normal limits. We conclude that closely monitored cyclosporine therapy may be a safe and effective treatment in patients with severe proteinuria and Alport syndrome. PMID- 17130109 TI - The relation between left ventricular diastolic indices and serum ferritin in thalassemia major. AB - Iron-induced heart failure is the primary cause of death in thalassemia major patients who receive continuous transfusions. Recent studies have suggested that diastolic function is impaired prior to systolic function in process of hemochromatosis, but they did not come to agreement on the first impaired diastolic index. Additionally, serum ferritin concentration is not a reliable indicator of body iron storage since it increases in any simple inflammation. Accordingly, the authors undertook this study to assess any association between left ventricular diastolic indices and serum ferritin in thalassemic patients with normal systolic function to estimate the true amount of body iron storage and correct it in earlier stages. Serum ferritin concentration and diastolic indices were measured in 29 patients with normal left ventricular systolic function. Linear regression test was used to find any association between hematological and cardiac factors. No significant association was found between diastolic indices and serum ferritin concentration. But the results were quite different in patients above and below 15 years of age; standardized coefficients (r) for peak of E and A were increased in patients above 15, and the significance was close to .05, unlike those of younger group. Although no correlation was found between serum ferritin and diastolic indices, the results were noteworthy in patients above 15. To appropriately judge this relation, the study must be continued with a bigger sample size and having patients' mean serum ferritin concentration during the 2 past years. PMID- 17130110 TI - Nutritional vitamin B12 deficiency in hospitalized young children. AB - The authors sought to determine prevalence, social, economic, and dietary patterns of young children (n = 20) identified as having vitamin B12 deficiency anemia after admission to their hospital in the last 3 years. The diagnosis of vitamin B12 deficiency was based on symptoms and clinical findings, findings on peripheral blood films and bone marrow aspirates, and serum levels of vitamin B12. The children had been exclusively breast-fed without any animal food supplementation. Serum vitamin B12 levels were also measured in the sera of mothers and found to be low. The authors concluded that vitamin B12 deficiency might be an important health problem among children of mothers who do not consume animal foods adequately. PMID- 17130111 TI - T-lymphocyte production, function, and death in children who recovered from opsoclonus-myoclonus syndrome. AB - Reduced CD4+ lymphocytes have been recently found in peripheral blood of children with active opsoclonus-myoclonus syndrome. The authors identified 2 children who recovered from this syndrome, one of whom showed reduced CD4+ lymphocytes 2 years after the disease onset. Except for a decrease of "naive" CD45RA+ CD4+ population and a mild restriction of T-cell heterogeneity in this patient, probably related to the immune response to viral infections, no alterations of T-cell homeostasis and function were found in either child. Therefore, the decrease of CD4+ cells may persist after clinical recovery, but the causes of this abnormality cannot be ascribed to intrinsic T-cell defects. PMID- 17130112 TI - Ovarian tissue cryostorage and grafting: an option to preserve fertility in pediatric patients with malignancies. AB - Fertility preservation in childhood cancer has become an important area of investigation due to increasing survival rates after cancer therapy. For these patients with an increased risk of infertility and premature ovarian failure, cryopreservation of ovarian tissue is a promising tool to preserve at least part of the reproductive potential. In recent years significant improvements have been achieved in this area, and 2 live births after autografting of frozen-thawed ovarian tissue have been reported. However, further research is needed to assess the clinical effectiveness of ovarian cryopreservation, to optimize the technique, and to limit the risk of reintroducing cancer cells in the patient with the graft. PMID- 17130113 TI - Nail involvement in langerhans cell histiocytosis. AB - Nail involvement, which is distinctly uncommon in Langerhans cell histiocytosis (LCH), is characterized by various features like longitudinal grooving, purpuric striae, hyperkeratosis, subungual pustules, deformity, loss of nail plate, paronychia, onycholysis, and pitting. Here the authors report the case of a child who presented with isolated nail unit changes due to Langerhans cell histiocytosis before the evolution of systemic features. The authors suggest that LCH should be considered in differential diagnoses of nail changes that are resistant to therapy. PMID- 17130114 TI - Primary intraspinal primitive neuroectodermal tumor treated with autologous stem cell transplantation: case report and review of the literature. AB - The authors describe the case of a 19-year-old female patient with a primary primitive neuroectodermal tumor (PNET) of the thoraco-lumbar spinal cord, who presented with acute urinary retention and back pain for 2 months. Magnetic resonance imaging revealed an intradural extramedullary tumor, 6.5 cm long, in the region of the conus medullaris. Histological examination disclosed a small round cell tumor with immunohistochemical characteristics of a peripheral PNET. Metastatic workup showed no evidence of an intracranial tumor or metastases outside the neuroaxis. The patient received multidisciplinary treatment, including surgical excision, irradiation of the entire cranio-spinal axis, and high-dose chemotherapy with autologous stem cell rescue. Presently, 24 months after diagnosis, the patient remains in complete remission. PMID- 17130115 TI - Invasive esophageal aspergillosis associated with acute myelogenous leukemia: successful therapy with combination caspofungin and liposomal amphotericin B. AB - Aspergillosis is one of the most common invasive fungal infections in patients with leukemia. In this patient group, this form of Aspergillus infection is a life-threatening condition with a mortality of 50-100%. The lungs are most often affected, but the esophagus can also be involved.The authors report the case of a child with leukemia who developed invasive esophageal aspergillosis. The condition was diagnosed by microscopic examination of endoscopic biopsy specimens. The patient was already receiving empirical liposomal amphotericin B when the diagnosis was made, so a second antifungal (caspofungin) was added to the regimen. This combination was successful. This case to demonstrates a case of successful treatment of invasive esophageal aspergillosis using combination therapy of liposomal amphotericin B and caspofungin. PMID- 17130116 TI - Use of recombinant erythropoietin for the management of severe hemolytic disease of the newborn of a K0 phenotype mother. AB - Very few people do not express any Kell antigens on their red blood cells (K0 phenotype). They can be immunized by transfusion or pregnancy and develop antibodies against Kell system antigens. These maternal antibodies can cause severe hemolytic disease of the fetus/newborn, as a result of the suppression of erythropoiesis and hemolysis. Multiple intrauterine transfusions in the management of severe hemolytic disease have been shown to cause erythropoietic suppression as well. Recombinant erythropoietin has been successfully used in the management of late anemia of infants with Rh hemolytic disease and in 1 case of KEL1 (Kell)-associated hemolytic disease. The authors present the case of severe hemolytic disease of a newborn due to KEL5 (Ku) isoimmunization of his K0 phenotype mother. Regular intrauterine transfusions were performed to manage the severe fetal anemia (Hb 3 g/dL). A male infant was born at the 36th week of gestation having normal hemoglobin (15.8 g/dL) and developed only mild hyperbilirubinemia. On the 15th day of life, the infant's hematocrit had fallen to 27.3%, with low reticulocyte count and low erythropoietin level. The infant was managed successfully with recombinant erythropoietin. PMID- 17130117 TI - Surviving with thalassemia major: the Italian experience. PMID- 17130118 TI - Progress in the treatment of childhood brain tumors: no room for complacency. PMID- 17130119 TI - Congenital neuroblastoma with multiple subcutaneous noduls. PMID- 17130120 TI - Phospholipid flippases. PMID- 17130121 TI - Myostatin induces cyclin D1 degradation to cause cell cycle arrest through a phosphatidylinositol 3-kinase/AKT/GSK-3 beta pathway and is antagonized by insulin-like growth factor 1. AB - Myostatin is a transforming growth factor beta superfamily member and is known as an inhibitor of skeletal muscle cell proliferation and differentiation. Exposure to myostatin induces G1 phase cell cycle arrest. In this study, we demonstrated that myostatin down-regulates Cdk4 activity via promotion of cyclin D1 degradation. Overexpression of cyclin D1 significantly blocked myostatin-induced proliferation inhibition. We further showed that phosphorylation at threonine 286 by GSK-3beta was required for myostatin-stimulated cyclin D1 nuclear export and degradation. This process is dependent upon the activin receptor IIB and the phosphatidylinositol 3-kinase/Akt pathway but not Smad3. Insulin-like growth factor 1 (IGF-1) treatment or Akt activation attenuated the myostatin-stimulated cyclin D1 degradation as well as the associated cell proliferation repression. In contrast, attenuation of IGF-1 signaling caused C2C12 cells to undergo apoptosis in response to myostatin treatment. The observation that IGF-1 treatment increases myostatin expression through a phosphatidylinositol 3-kinase pathway suggests a possible feedback regulation between IGF-1 and myostatin. These findings uncover a novel role for myostatin in the regulation of cell growth and cell death in concert with IGF-1. PMID- 17130122 TI - Mutations in the SF1-U2AF59-U2AF23 complex cause exon skipping in Schizosaccharomyces pombe. AB - To identify genes involved in the mechanism to ensure ordered 5' to 3' exon joining in constitutively spliced pre-mRNAs, we screened for mutants that cause exon skipping in the fission yeast Schizosaccharomyces pombe using a reporter plasmid, which contains the ura4+ gene with the nda3 intron 1-exon 2-intron 2 sequence. The reporter plasmid was designed to produce the functional ura4+ mRNA, when the central nda3 exon is skipped during the splicing reaction. We mutagenized cells harboring the plasmid by UV irradiation and isolated 34 ura+ mutants that grew on minimal medium. Of those, eight mutants were found to be temperature sensitive (ts) for growth. Complementation analyses revealed that the ts mutants belong to three distinct complementation groups named ods (ordered splicing) 1, 2, and 3. RT-PCR analyses showed that products of exon skipping were actually generated in the ods mutants. We cloned the genes responsible for the ods mutations, and found that ods1+, ods2+, and ods3+ encode splicing factors Prp2p/U2AF59, U2AF23, and SF1, respectively, which form a SF1-U2AF59-U2AF23 complex involved in recognition of the branch-point and 3' splice site sequences in a pre-mRNA. We also showed that mutations in the SF1-U2AF59-U2AF23 binding sequences in the reporter plasmid result in exon skipping in wild-type S. pombe cells. In addition, drugs that decrease the rate of transcription elongation were found to suppress the exon skipping in the ods mutants. These results suggest that co-transcriptional recognition of a nascent pre-mRNA by the SF1-U2AF59 U2AF23 complex is essential for ordered exon joining in constitutive splicing in S. pombe. PMID- 17130123 TI - Lysyl oxidase interacts with hormone placental lactogen and synergistically promotes breast epithelial cell proliferation and migration. AB - Lysyl oxidase (LOX), an extracellular amine oxidase, catalyzes the cross-linking of collagen and elastin. LOX has been also shown to play an essential role in promoting the invasive and metastatic potential of breast tumor cells. However, the LOX-interacting factors in these processes are not known. In this study, we identified placental lactogen (PL), a member of the growth hormone/prolactin hormone family, as a LOX-interacting partner using yeast two-hybrid screens. PL is normally only expressed in placental syncytiotrophoblasts, but PL genes are amplified and expressed in a high percentage of invasive ductal breast carcinomas. We confirmed LOX-PL interactions using far Western and solid phase binding assays. In activity assays, PL was not a substrate or inhibitor of LOX. We further demonstrated that PL is expressed in breast tumor epithelial cells and detected LOX-PL interactions by coimmunoprecipitation in invasive breast cancer cells. In MCF-10A normal breast epithelial cells stably expressing LOX, PL, or both, LOX had no effect on cell proliferation, PL alone increased proliferation by 49%, and coexpression of LOX and PL led to a 121% increase in cell proliferation. Unlike in tumor cells, LOX did not induce a more migratory phenotype in MCF-10A cells; nor did PL. However, their coexpression resulted in a 240% increase in cell migration, suggesting that these interactions may be highly relevant to the transition of epithelial cells toward a migratory phenotype during the development and progression of breast carcinoma and a significant role for LOX-PL interactions in epithelial cell behavior. PMID- 17130124 TI - A novel, non-immunogenic Fyn SH3-derived binding protein with tumor vascular targeting properties. AB - The generation of novel binding molecules based on protein frameworks ("scaffolds") represents an emerging field in protein engineering, with the potential to replace antibodies for many research and clinical applications. Here, we describe the design, construction, characterization, and use of a novel human Fyn SH3 phage library, containing 1.2 x 10(9) individual clone members. We also present the isolation and in vitro characterization of Fyn SH3-derived proteins binding to the extra-domain B of fibronectin, a marker of angiogenesis. One specific binding clone, named D3, was further evaluated and showed a remarkable ability to stain vascular structures in tumor sections. Furthermore, quantitative biodistribution studies in tumor-bearing mice revealed the ability of D3 to selectively accumulate in the tumor. In contrast to human scFv antibody fragments administered to mice, neither Fyn SH3 WT nor the D3 mutant was immunogenic in mice after four intravenous injections. The extra-domain B binding D3 protein opens new biomedical opportunities for the in vivo imaging of solid tumors and for the delivery of toxic agents to the tumoral vasculature. PMID- 17130125 TI - Structure of Aquifex aeolicus argonaute highlights conformational flexibility of the PAZ domain as a potential regulator of RNA-induced silencing complex function. AB - Gene silencing mediated by RNA interference requires the sequence-specific recognition of target mRNA by the endonuclease Argonaute, the primary enzymatic component of the RNA-induced silencing complex. We report the crystal structure of Aquifex aeolicus Argonaute, refined at 3.2A resolution. Relative to recent Argonaute structures, a 24 degrees reorientation of the PAZ domain in our structure opens a basic cleft between the N-terminal and PAZ domains, exposing the guide strand binding pocket of PAZ. This rearrangement leads to a branched, Y shaped system of grooves that extends through the molecule and merges in a central channel containing the catalytic residues. A 5.5-ns molecular dynamics simulation of Argonaute shows a strong tendency of the PAZ and N-terminal domains to be mobile. Binding of single-stranded DNA to Argonaute monitored by total internal reflection fluorescence spectroscopy shows biphasic kinetics, also indicative of domain rearrangement upon DNA binding. Conformational rearrangement of the PAZ domain may therefore be critical for the catalytic cycle of Argonaute and the RNA-induced silencing complex. PMID- 17130126 TI - Chaperone properties of mammalian mitochondrial translation elongation factor Tu. AB - The main function of the prokaryotic translation elongation factor Tu (EF-Tu) and its eukaryotic counterpart eEF1A is to deliver aminoacyl-tRNA to the A-site on the ribosome. In addition to this primary function, it has been reported that EF Tu from various sources has chaperone activity. At present, little information is available about the chaperone activity of mitochondrial EF-Tu. In the present study, we have examined the chaperone function of mammalian mitochondrial EF-Tu (EF-Tumt). We demonstrate that recombinant EF-Tumt prevents thermal aggregation of proteins and enhances protein refolding in vitro and that this EF-Tumt chaperone activity proceeds in a GTP-independent manner. We also demonstrate that, under heat stress, the newly synthesized peptides from the mitochondrial ribosome specifically co-immunoprecipitate with EF-Tumt and are destabilized in EF-Tumt-overexpressing cells. We show that most of the EF-Tumt localizes on the mitochondrial inner membrane where most mitochondrial ribosomes are found. We discuss the possible role of EF-Tumt chaperone activity in protein quality control in mitochondria, with regard to the recently reported in vivo chaperone function of eEF1A. PMID- 17130127 TI - Spectropotentiometric and structural analysis of the periplasmic nitrate reductase from Escherichia coli. AB - The Escherichia coli NapA (periplasmic nitrate reductase) contains a [4Fe-4S] cluster and a Mo-bis-molybdopterin guanine dinucleotide cofactor. The NapA holoenzyme associates with a di-heme c-type cytochrome redox partner (NapB). These proteins have been purified and studied by spectropotentiometry, and the structure of NapA has been determined. In contrast to the well characterized heterodimeric NapAB systems ofalpha-proteobacteria, such as Rhodobacter sphaeroides and Paracoccus pantotrophus, the gamma-proteobacterial E. coli NapA and NapB proteins purify independently and not as a tight heterodimeric complex. This relatively weak interaction is reflected in dissociation constants of 15 and 32 mum determined for oxidized and reduced NapAB complexes, respectively. The surface electrostatic potential of E. coli NapA in the apparent NapB binding region is markedly less polar and anionic than that of the alpha-proteobacterial NapA, which may underlie the weaker binding of NapB. The molybdenum ion coordination sphere of E. coli NapA includes two molybdopterin guanine dinucleotide dithiolenes, a protein-derived cysteinyl ligand and an oxygen atom. The Mo-O bond length is 2.6 A, which is indicative of a water ligand. The potential range over which the Mo(6+) state is reduced to the Mo(5+) state in either NapA (between +100 and -100 mV) or the NapAB complex (-150 to -350 mV) is much lower than that reported for R. sphaeroides NapA (midpoint potential Mo(6+/5+) > +350 mV), and the form of the Mo(5+) EPR signal is quite distinct. In E. coli NapA or NapAB, the Mo(5+) state could not be further reduced to Mo(4+). We then propose a catalytic cycle for E. coli NapA in which nitrate binds to the Mo(5+) ion and where a stable des-oxo Mo(6+) species may participate. PMID- 17130128 TI - Nutlin-3 protects kidney cells during cisplatin therapy by suppressing Bax/Bak activation. AB - Nutlins, the newly developed small molecule antagonists of MDM2, activate p53 and induce apoptosis in cancer cells, offering a novel strategy of chemotherapy. Recent studies have further suggested synergistic effects of nutlins with other chemotherapeutic drugs. However, it is unclear whether nutlins increase or decrease the side effects of these drugs in normal non-malignant cells or tissues. Cisplatin is a widely used chemotherapy drug, which has a major side effect of kidney injury. Here we show that Nutlin-3 protected kidney cells against cisplatin-induced apoptosis. The cytoprotective effects of Nutlin-3 were not related to its regulation of p53 or consequent gene expression during cisplatin treatment. Moreover, the protective effects were shown in MDM2-, MDM4-, or p53-deficient cells. On the other hand, Nutlin-3 suppressed mitochondrial events of apoptosis during cisplatin incubation, including Bax activation and cytochrome c release. Nutlin-3 attenuated cisplatin-induced oligomerization of Bax and Bak but not their interactions with Bcl-XL. In isolated mitochondria, Nutlin-3 inhibited cytochrome c release induced by Ca2+, Bim peptide, and recombinant tBid. Importantly, it blocked both Bax and Bak oligomerization under these conditions. Together, the results have uncovered a new pharmacological function of nutlins, i.e. suppression of Bax and Bak, two critical mediators of apoptosis. PMID- 17130129 TI - Thioredoxin-dependent enzymatic activation of mercaptopyruvate sulfurtransferase. An intersubunit disulfide bond serves as a redox switch for activation. AB - Rat 3-mercaptopyruvate sulfurtransferase (MST) contains three exposed cysteines as follows: a catalytic site cysteine, Cys(247), in the active site and Cys(154) and Cys(263) on the surface of MST. The corresponding cysteine to Cys(263) is conserved in mammalian MSTs, and Cys(154) is a unique cysteine. MST has monomer dimer equilibrium with the assistance of oxidants and reductants. The monomer to dimer ratio is maintained at approximately 92:8 in 0.2 m potassium phosphate buffer containing no reductants under air-saturated conditions; the dimer might be symmetrical via an intersubunit disulfide bond between Cys(154) and Cys(154) and between Cys(263) and Cys(263), or asymmetrical via an intersubunit disulfide bond between Cys(154) and Cys(263). Escherichia coli reduced thioredoxin (Trx) cleaved the intersubunit disulfide bond to activate MST to 2.3- and 4.9-fold the levels of activation of dithiothreitol (DTT)-treated and DTT-untreated MST, respectively. Rat Trx also activated MST. On the other hand, reduced glutathione did not affect MST activity. E. coli C35S Trx, in which Cys(35) was replaced with Ser, formed some adducts with MST and activated MST after treatment with DTT. Thus, Cys(32) of E. coli Trx reacted with the redox-active cysteines, Cys(154) and Cys(263), by forming an intersubunit disulfide bond and a sulfenyl Cys(247). A consecutively formed disulfide bond between Trx and MST must be cleaved for the activation. E. coli C32S Trx, however, did not activate MST. Reduced Trx turns on a redox switch for the enzymatic activation of MST, which contributes to the maintenance of cellular redox homeostasis. PMID- 17130130 TI - The role of activating protein 1 in the transcriptional regulation of the human FCGR2B promoter mediated by the -343 G -> C polymorphism associated with systemic lupus erythematosus. AB - The inhibitory receptor FcgammaRIIb is a negative regulator of antibody production and inflammatory responses. The -343 G --> C polymorphism in the human FCGR2B promoter is associated with systemic lupus erythematosus. The -343 C mutant promoter has decreased transcriptional activity. In the present study, we show that the transcriptional change correlates with quantitative differences in the interaction of the activating protein 1 complex with the mutant FCGR2B promoter. Promoter pulldown and chromatin immunoprecipitation assays demonstrated binding of c-Jun to the FCGR2B promoter. Phosphorylation of c-Jun was accompanied by transactivation of both FCGR2B promoter variants, whereas dephosphorylation of c-Jun by an inhibitor of c-Jun N-terminal kinase, markedly decreased the promoter activities. The -343 G --> C substitution enabled the specific interaction of the transcription factor Yin-Yang 1 with the mutant FCGR2B promoter. Yin-Yang 1 competed with activating protein 1 for binding at the -343 site, and contributed to the repression of the mutant FCGR2B promoter activity. This mechanism could be responsible for the decreased expression of FcgammaRIIb associated with the -343 C/C homozygous FCGR2B genotype in lupus patients. These findings provide a rationale for the transcriptional defect mediated by the -343 C/C FCGR2B promoter polymorphism associated with systemic lupus erythematosus, and add to our understanding of the complex transcriptional regulation of the human FCGR2B promoter. PMID- 17130131 TI - Cellular adhesion responses to the heparin-binding (HepII) domain of fibronectin require heparan sulfate with specific properties. AB - Cell surface heparan sulfate (HS) proteoglycans are required in development and postnatal repair. Important classes of ligands for HS include growth factors and extracellular matrix macromolecules. For example, the focal adhesion component syndecan-4 interacts with the III(12-14) region of fibronectin (HepII domain) through its HS chains. The fine structure of HS is critical to growth factor responses, and whether this extends to matrix ligands is unknown but is suggested from in vitro experiments. Cell attachment to HepII showed that heparin oligosaccharides of >or=14 sugar residues were required for optimal inhibition. The presence of N-sulfated glucosamine in the HS was essential, whereas 2-O sulfation of uronic acid or 6-O-sulfation of glucosamine had marginal effects. In the more complex response of focal adhesion formation through syndecan-4, N sulfates were again required and also glucosamine 6-O-sulfate. The significance of polymer N-sulfation and sulfated domains in HS was confirmed by studies with mutant Chinese hamster ovary cells where heparan sulfation was compromised. Finally, focal adhesion formation was absent in fibroblasts synthesizing short HS chains resulting from a gene trap mutation in one of the two major glucosaminoglycan polymerases (EXT1). Several separate, specific properties of cell surface HS are therefore required in cell adhesion responses to the fibronectin HepII domain. PMID- 17130132 TI - Functional analysis of individual binding activities of the scaffold protein eIF4G. AB - Eukaryotic initiation factor (eIF) 4G is an integral member of the translation initiation machinery. The molecule serves as a scaffold for several other initiation factors, including eIF4E, eIF4AI, the eIF3 complex, and poly(A) binding protein (PABP). Previous work indicates that complexes between these proteins exhibit enhanced mRNA cap-binding and RNA helicase activities relative to the respective individual proteins, eIF4E and eIF4A. The eIF4G-PABP interaction has been implicated in enhancing the formation of 48 S and 80 S initiation complexes and ribosome recycling through mRNA circularization. The eIF3-eIF4GI interaction is believed to forge the link between the 40 S subunit and the mRNA. Here we have investigated the behavior in vitro and in intact cells of eIF4GIf molecules lacking either the PABP-binding site, the eIF3-binding site, the middle domain eIF4A-binding site, or the C-terminal segment that includes the second eIF4A-binding site. Although in some cases the mutant forms were recruited more slowly, all of these eIF4G variants could form complexes with eIF4E, enter 48 S complexes and polysomes in vivo and in vitro, and partially rescue translation in cells targeted with eIF4GI short interfering RNA. In the reticulocyte lysate, eIF4G unable to interact directly with PABP showed little impairment in its ability to support translation, whereas loss of either of the eIF4A-binding sites or the eIF3-binding site resulted in a marked decrease in activity. We conclude that there is considerable redundancy in the mechanisms forming initiation complexes in mammalian cells, such that many individual interactions have regulatory rather than essential roles. PMID- 17130133 TI - The slow skeletal muscle isoform of myosin shows kinetic features common to smooth and non-muscle myosins. AB - Fast and slow mammalian muscle myosins differ in the heavy chain sequences (MHC 2, MHC-1) and muscles expressing the two isoforms contract at markedly different velocities. One role of slow skeletal muscles is to maintain posture with low ATP turnover, and MHC-1 expressed in these muscles is identical to heavy chain of the beta-myosin of cardiac muscle. Few studies have addressed the biochemical kinetic properties of the slow MHC-1 isoform. We report here a detailed analysis of the MHC-1 isoform of the rabbit compared with MHC-2 and focus on the mechanism of ADP release. We show that MHC-1, like some non-muscle myosins, shows a biphasic dissociation of actin-myosin by ATP. Most of the actin-myosin dissociates at up to approximately 1000 s(-1), a very similar rate constant to MHC-2, but 10-15% of the complex must go through a slow isomerization (approximately 20 s(-1)) before ATP can dissociate it. Similar slow isomerizations were seen in the displacement of ADP from actin-myosin.ADP and provide evidence of three closely related actin myosin.ADP complexes, a complex in rapid equilibrium with free ADP, a complex from which ADP is released at the rate required to define the maximum shortening velocity of slow muscle fibers (approximately 20 s(-1)), and a third complex that releases ADP too slowly (approximately 6 s(-1)) to be on the main ATPase pathway. The role of these actin-myosin.ADP complexes in the mechanochemistry of slow muscle contraction is discussed in relation to the load dependence of ADP release. PMID- 17130134 TI - Calmodulin-like protein increases filopodia-dependent cell motility via up regulation of myosin-10. AB - Human calmodulin-like protein (CLP) is an epithelial-specific protein that is expressed during cell differentiation but down-regulated in primary cancers and transformed cell lines. Using stably transfected and inducible HeLa cell lines, we found that CLP expression did not alter the proliferation rate and colony forming potential of these cells. However, remarkable phenotypic changes were observed in CLP-expressing compared with control cells. Soft agar colonies of CLP expressing cells had rough boundaries, with peripheral cells migrating away from the colony. Cells expressing CLP displayed a striking increase in the number and length of myosin-10-positive filopodia and showed increased mobility in a wound healing assay. This increase in wound healing capacity was prevented by small interference RNA-mediated down-regulation of myosin-10. Fluorescence microscopy and Western blotting revealed that CLP expression results in up-regulation of its target protein, myosin-10. This up-regulation occurs at the protein level by stabilization of myosin-10. Thus, CLP functions by increasing the stability of myosin-10, leading to enhanced myosin-10 function and a subsequent increase in filopodial dynamics and cell migration. In stratified epithelia, CLP may be required during terminal differentiation to increase myosin-10 function as cells migrate toward the upper layers and establish new adhesive contacts. PMID- 17130135 TI - MNK1 and MNK2 regulation in HER2-overexpressing breast cancer lines. AB - MAPK-interacting protein kinases 1 and 2 (MNK1 and MNK2) function downstream of p38 and ERK MAPK, but there are large gaps in our knowledge of how MNKs are regulated and function. As proteins activated in the HER2/Ras/Raf/ERK pathway, the MNKs are of potential interest in HER2-overexpressing cancers. We utilized a panel of breast cell lines (HCC1419, AU565, SKBR3, MCF7, and MCF10A), three of which overexpress HER2, to characterize the amounts and activation status of MNKs and other pathway enzymes (ERKs and RSKs) in these cells. We generated a phosphospecific antibody to Thr(P)-214 in the T-loop of MNKs and found that phosphorylations of both Thr-209 and Thr-214 in human MNK1 are required for activation. Increased phosphorylation and activity of the MNKs correlate with HER2 overexpression, and inhibition of the MNKs reduces colony formation in soft agar. Our work identifies the MNKs as potential therapeutic targets for breast cancer treatments. PMID- 17130136 TI - Regulation of human melanocortin 1 receptor signaling and trafficking by Thr-308 and Ser-316 and its alteration in variant alleles associated with red hair and skin cancer. AB - The melanocortin 1 receptor (MC1R), a G protein-coupled receptor (GPCR) positively coupled to adenylyl cyclase, is a key regulator of melanocyte proliferation and differentiation and a determinant of pigmentation, skin phototype, and skin cancer risk. MC1R activation stimulates melanogenesis and increases the ratio of black, strongly photoprotective eumelanins to yellowish and poorly photoprotective pheomelanin pigments. Desensitization and internalization are key regulatory mechanisms of GPCR signaling. Agonist-induced desensitization usually depends on phosphorylation by a GPCR kinase (GRK) followed by receptor internalization in endocytic vesicles. We have shown that MC1R desensitization is mediated by two GRKs expressed in melanocytes and melanoma cells, GRK2 and GRK6. Here we show that in contrast with this dual specificity for desensitization, GRK6 but not GRK2 mediated MC1R internalization. Mutagenesis studies suggested that the targets of GRK6 are two residues located in the MC1R cytosolic C terminus, Thr-308 and Ser-316. A T308D/S316D mutant mimicking their phosphorylated state was constitutively desensitized and associated with endosomes, whereas a T308A/S316A mutant was resistant to desensitization and internalization. We studied the desensitization and internalization of three variant MC1R forms associated with red hair and increased skin cancer risk: R151C, R160W, and D294H. These variants showed a less efficient desensitization. Moreover, D294H was resistant to internalization, thus accounting for its abnormally high surface expression. Co-expression of variant and wild type MC1R modified its desensitization and internalization behavior. These data suggest that MC1R might be regulated by novel mechanisms including differential effects of GRKs and altered desensitization rates of certain allelic combinations. PMID- 17130137 TI - Improved breast cancer prognosis through the combination of clinical and genetic markers. AB - MOTIVATION: Accurate prognosis of breast cancer can spare a significant number of breast cancer patients from receiving unnecessary adjuvant systemic treatment and its related expensive medical costs. Recent studies have demonstrated the potential value of gene expression signatures in assessing the risk of post surgical disease recurrence. However, these studies all attempt to develop genetic marker-based prognostic systems to replace the existing clinical criteria, while ignoring the rich information contained in established clinical markers. Given the complexity of breast cancer prognosis, a more practical strategy would be to utilize both clinical and genetic marker information that may be complementary. METHODS: A computational study is performed on publicly available microarray data, which has spawned a 70-gene prognostic signature. The recently proposed I-RELIEF algorithm is used to identify a hybrid signature through the combination of both genetic and clinical markers. A rigorous experimental protocol is used to estimate the prognostic performance of the hybrid signature and other prognostic approaches. Survival data analyses is performed to compare different prognostic approaches. RESULTS: The hybrid signature performs significantly better than other methods, including the 70-gene signature, clinical makers alone and the St. Gallen consensus criterion. At the 90% sensitivity level, the hybrid signature achieves 67% specificity, as compared to 47% for the 70-gene signature and 48% for the clinical makers. The odds ratio of the hybrid signature for developing distant metastases within five years between the patients with a good prognosis signature and the patients with a bad prognosis is 21.0 (95% CI:6.5-68.3), far higher than either genetic or clinical markers alone. AVAILABILITY: The breast cancer dataset is available at www.nature.com and Matlab codes are available upon request. PMID- 17130138 TI - Hydroxyethyl starch is superior to lactated Ringer as a replacement fluid in a pig model of acute normovolaemic haemodilution. AB - BACKGROUND: Tissue tolerance to oxygen privation during acute normovolaemic haemodilution with different fluids remains unclear. We tested the hypothesis that hydroxyethyl starch (HES) is superior to lactated Ringer's solution in pigs for preserving tissue perfusion during acute normovolaemic haemodilution. METHODS: Twenty-four animals were randomized into control, lactated Ringer's solution and HES groups. All groups, except the control, underwent acute normovolaemic haemodilution. Haemodynamics, oxygen parameter indices, global anaerobic metabolic markers, echocardiographic parameters, gastric tonometry and serum osmolarity were monitored at baseline, immediately after (0 min) and 60 and 120 min after the end of haemodilution. Myocardial, liver, stomach and intestine samples were collected for further evaluation. RESULTS: Cardiac and oxygen parameter index responses to acute normovolaemic haemodilution were comparable. However, the increment in cardiac index, stroke volume index, and left ventricular stroke work index were more sustained in the starch group. In the lactated Ringer's group, gastric pH decreased significantly and was accompanied by a significant increase in lactate. Myocardial ultrastructure was better preserved in the starch group. The other tissue samples presented no change. CONCLUSIONS: In this model of ANH, the starch group had a superior haemodynamic response. Minor loss of myocardial cellular integrity and preserved gastric pHi reinforce these findings. PMID- 17130139 TI - Screening and brief intervention online for college students: the ihealth study. AB - AIMS: To test the feasibility of online alcohol screening and brief intervention (BI) by comparing (i) two approaches to inviting all students to be screened, and (ii) a minimal versus a more extensive BI. METHODS: Freshmen students at one university were randomized to receive one of two types of email invitations to an online anonymous: (i) general health assessment, or (ii) alcohol-specific assessment. All were linked to the same alcohol screening survey. Those with unhealthy alcohol use (AUDIT >or=8) were randomly assigned to minimal or more extensive online alcohol BI. RESULTS: In both invitation groups (4008 students), 55% of students completed the online screening. Overall, 37% of men and 26% of women had unhealthy alcohol use. Compared to minimal BI, more extensive BI was associated with intention to seek help among men and with a greater increase in readiness to change among women. One month after BI, 75% of students completed another assessment, 33% of women and 15% of men with unhealthy alcohol use at baseline no longer had unhealthy alcohol use. There were no significant differences on drinking measures by BI randomization group. CONCLUSIONS: Over half of an entire freshman class of college students were reached by email and completed alcohol screening and brief intervention. Even an alcohol-specific invitation did not deter students. Although brief interventions that differed had some gender specific effects on readiness to change and intention, in general, unhealthy alcohol use decreased after brief intervention. Web screening and brief intervention show promise for addressing unhealthy alcohol use by college students. PMID- 17130140 TI - HIV outbreak among injecting drug users in the Helsinki region: social and geographical pockets. AB - BACKGROUND: Incidence of newly diagnosed HIV infections among injecting drug users (IDUs) in Helsinki rose from 0 per 100,000 inhabitants in 1997 to 2.9 in 1998 and to 11.1 in 1999. Thereafter incidence declined to 2.1 in 2003. METHODS: Data were collected from interviews with HIV-positive IDUs who attended the University Hospital in Helsinki from 1998 until 2003. We studied the sociodemographic profile and spatial distribution of IDUs who were diagnosed in the beginning of the outbreak and those diagnosed later. The indicator for the spatial differentiation within the metropolitan area is % employed males aged 25 64. RESULTS: The outbreak occurred among a marginalized population of IDUs characterized by a long history of injecting drug use (10.7 years), mean age 32 years, homelessness (66.3%), history of imprisonment (74.7%) and psychiatric hospital care (40.6%). Compared with 98 early cases diagnosed during the first 2 years until 2000, 47 recent cases diagnosed after 2001 were 4 years older, and as marginalized. Except for the city centre, both early and recent cases had been living or using drugs in the same deprived neighbourhoods with the highest unemployment rates. Up to 40% of cases in the two big geographical clusters did not have contact with the city centre, where the needle exchange services were available. CONCLUSIONS: The Finnish HIV outbreak is restricted socially to a very marginalized IDU population, and spatially to local pockets of poverty. In low prevalence countries, prevention programs should be targeted early at high-risk areas and populations. PMID- 17130141 TI - Socioeconomic status, psychosocial health and health behaviours among Hungarian adolescents. AB - BACKGROUND: While socioeconomic differences in health, morbidity, and disability are highest among middle-aged persons, there is a certain level of 'equalization' during adolescence and young adulthood. Despite this equalization, however, there still are differences in psychosocial variables or health-related behaviours, often very subtle and sometimes difficult to measure. METHODS: Using data (n = 1114) on high school students (aged between 14 and 21 years) from the Southern Plain Region, Hungary, the present study looks at the role of multiple SES indicators (objective and subjective; occupation and education; family structure) in adolescents' psychosocial health (self-perceived health, psychosomatic, and depressive symptomatology) and health behaviour (substance use and sports activity). RESULTS: Based on the results of multivariate logistic regression analyses, findings suggest the following: (i) SES self-assessment proved to be a significant predictor of adolescents' psychosocial health and health behaviours; (ii) family structure (that is, living in a non-intact family) also significantly influenced adolescents' psychosocial health and health behaviours; (iii) parents' employment status and schooling had a limited influence on their children's health outcomes; (iv) in a word, SES gradients in adolescents' psychosocial health and health behaviour were inconsistent and sometimes irregular (that is, inverse). The subjective SES measurement plays an important role (positive association), whereas certain types of parents' inactive status (in terms of labour market, that is, unemployment or retirement) seem to act in a predictable way (negative association). CONCLUSIONS: Our results indicate that despite certain level of equalization during adolescence, some important relationships between SES variables and health outcomes may occur. PMID- 17130142 TI - ForestTreeDB: a database dedicated to the mining of tree transcriptomes. AB - ForestTreeDB is intended as a resource that centralizes large-scale expressed sequence tag (EST) sequencing results from several tree species (http://foresttree.org/ftdb). It currently encompasses 344,878 quality sequences from 68 libraries, from diverse organs of conifer and hybrid poplar trees. It utilizes the Nimbus data model to provide a hosting system for multiple projects, and uses object-relational mapping APIs in Java and Perl for data accesses within an Oracle database designed to be scalable, maintainable and extendable. Transcriptome builds or unigene sets occupy the focal point of the system. Several of the five current species-specific unigenes were used to design microarrays and SNP resources. The ForestTreeDB web application provides the means for multiple combination database queries. It presents the user with a list of discrete queries to retrieve and download large EST datasets or sequences from precompiled unigene assemblies. Functional annotation assignment is not trivial in conifers which are distantly related to angiosperm model plants. Optimal annotations are achieved through database queries that integrate results from several procedures based open-source tools. ForestTreeDB aims to facilitate sequence mining of coherent annotations in multiple species to support comparative genomic approaches. We plan to continuously enrich ForestTreeDB with other resources through collaborations with other genomic projects. PMID- 17130143 TI - HepSEQ: International Public Health Repository for Hepatitis B. AB - HepSEQ is a repository for an extensive library of public health and molecular data relating to hepatitis B virus (HBV) infection collected from international sources. It is hosted by the Centre for Infections, Health Protection Agency (HPA), England, United Kingdom. This repository has been developed as a web enabled, quality-controlled database to act as a tool for surveillance, HBV case management and for research. The web front-end for the database system can be accessed from http://www.hpa-bioinfodatabases.org.uk/hepatitis_open/main.php. The format of the database system allows for comprehensive molecular, clinical and epidemiological data to be deposited into a functional database, to search and manipulate the stored data and to extract and visualize the information on epidemiological, virological, clinical, nucleotide sequence and mutational aspects of HBV infection through web front-end. Specific tools, built into the database, can be utilized to analyse deposited data and provide information on HBV genotype, identify mutations with known clinical significance (e.g. vaccine escape, precore and antiviral-resistant mutations) and carry out sequence homology searches against other deposited strains. Further mechanisms are also in place to allow specific tailored searches of the database to be undertaken. PMID- 17130144 TI - PANTHER version 6: protein sequence and function evolution data with expanded representation of biological pathways. AB - PANTHER is a freely available, comprehensive software system for relating protein sequence evolution to the evolution of specific protein functions and biological roles. Since 2005, there have been three main improvements to PANTHER. First, the sequences used to create evolutionary trees are carefully selected to provide coverage of phylogenetic as well as functional information. Second, PANTHER is now a member of the InterPro Consortium, and the PANTHER hidden markov Models (HMMs) are distributed as part of InterProScan. Third, we have dramatically expanded the number of pathways associated with subfamilies in PANTHER. Pathways provide a detailed, structured representation of protein function in the context of biological reaction networks. PANTHER pathways were generated using the emerging Systems Biology Markup Language (SBML) standard using pathway network editing software called CellDesigner. The pathway collection currently contains approximately 1500 reactions in 130 pathways, curated by expert biologists with authorship attribution. The curation environment is designed to be easy to use, and the number of pathways is growing steadily. Because the reaction participants are linked to subfamilies and corresponding HMMs, reactions can be inferred across numerous different organisms. The HMMs can be downloaded by FTP, and tools for analyzing data in the context of pathways and function ontologies are available at http://www.pantherdb.org. PMID- 17130145 TI - Michigan Molecular Interactions (MiMI): putting the jigsaw puzzle together. AB - Protein interaction data exists in a number of repositories. Each repository has its own data format, molecule identifier and supplementary information. Michigan Molecular Interactions (MiMI) assists scientists searching through this overwhelming amount of protein interaction data. MiMI gathers data from well known protein interaction databases and deep-merges the information. Utilizing an identity function, molecules that may have different identifiers but represent the same real-world object are merged. Thus, MiMI allows the users to retrieve information from many different databases at once, highlighting complementary and contradictory information. To help scientists judge the usefulness of a piece of data, MiMI tracks the provenance of all data. Finally, a simple yet powerful user interface aids users in their queries, and frees them from the onerous task of knowing the data format or learning a query language. MiMI allows scientists to query all data, whether corroborative or contradictory, and specify which sources to utilize. MiMI is part of the National Center for Integrative Biomedical Informatics (NCIBI) and is publicly available at: http://mimi.ncibi.org. PMID- 17130146 TI - SwissRegulon: a database of genome-wide annotations of regulatory sites. AB - SwissRegulon (http://www.swissregulon.unibas.ch) is a database containing genome wide annotations of regulatory sites in the intergenic regions of genomes. The regulatory site annotations are produced using a number of recently developed algorithms that operate on multiple alignments of orthologous intergenic regions from related genomes in combination with, whenever available, known sites from the literature, and ChIP-on-chip binding data. Currently SwissRegulon contains annotations for yeast and 17 prokaryotic genomes. The database provides information about the sequence, location, orientation, posterior probability and, whenever available, binding factor of each annotated site. To enable easy viewing of the regulatory site annotations in the context of other features annotated on the genomes, the sites are displayed using the GBrowse genome browser interface and can be queried based on any annotated genomic feature. The database can also be queried for regulons, i.e. sites bound by a common factor. PMID- 17130147 TI - H-DBAS: alternative splicing database of completely sequenced and manually annotated full-length cDNAs based on H-Invitational. AB - The Human-transcriptome DataBase for Alternative Splicing (H-DBAS) is a specialized database of alternatively spliced human transcripts. In this database, each of the alternative splicing (AS) variants corresponds to a completely sequenced and carefully annotated human full-length cDNA, one of those collected for the H-Invitational human-transcriptome annotation meeting. H-DBAS contains 38,664 representative alternative splicing variants (RASVs) in 11,744 loci, in total. The data is retrievable by various features of AS, which were annotated according to manual annotations, such as by patterns of ASs, consequently invoked alternations in the encoded amino acids and affected protein motifs, GO terms, predicted subcellular localization signals and transmembrane domains. The database also records recently identified very complex patterns of AS, in which two distinct genes seemed to be bridged, nested or degenerated (multiple CDS): in all three cases, completely unrelated proteins are encoded by a single locus. By using AS Viewer, each AS event can be analyzed in the context of full-length cDNAs, enabling the user's empirical understanding of the relation between AS event and the consequent alternations in the encoded amino acid sequences together with various kinds of affected protein motifs. H-DBAS is accessible at http://jbirc.jbic.or.jp/h-dbas/. PMID- 17130148 TI - NCBI reference sequences (RefSeq): a curated non-redundant sequence database of genomes, transcripts and proteins. AB - NCBI's reference sequence (RefSeq) database (http://www.ncbi.nlm.nih.gov/RefSeq/) is a curated non-redundant collection of sequences representing genomes, transcripts and proteins. The database includes 3774 organisms spanning prokaryotes, eukaryotes and viruses, and has records for 2,879,860 proteins (RefSeq release 19). RefSeq records integrate information from multiple sources, when additional data are available from those sources and therefore represent a current description of the sequence and its features. Annotations include coding regions, conserved domains, tRNAs, sequence tagged sites (STS), variation, references, gene and protein product names, and database cross-references. Sequence is reviewed and features are added using a combined approach of collaboration and other input from the scientific community, prediction, propagation from GenBank and curation by NCBI staff. The format of all RefSeq records is validated, and an increasing number of tests are being applied to evaluate the quality of sequence and annotation, especially in the context of complete genomic sequence. PMID- 17130149 TI - VISTA Enhancer Browser--a database of tissue-specific human enhancers. AB - Despite the known existence of distant-acting cis-regulatory elements in the human genome, only a small fraction of these elements has been identified and experimentally characterized in vivo. This paucity of enhancer collections with defined activities has thus hindered computational approaches for the genome-wide prediction of enhancers and their functions. To fill this void, we utilize comparative genome analysis to identify candidate enhancer elements in the human genome coupled with the experimental determination of their in vivo enhancer activity in transgenic mice [L. A. Pennacchio et al. (2006) Nature, in press]. These data are available through the VISTA Enhancer Browser (http://enhancer.lbl.gov). This growing database currently contains over 250 experimentally tested DNA fragments, of which more than 100 have been validated as tissue-specific enhancers. For each positive enhancer, we provide digital images of whole-mount embryo staining at embryonic day 11.5 and an anatomical description of the reporter gene expression pattern. Users can retrieve elements near single genes of interest, search for enhancers that target reporter gene expression to a particular tissue, or download entire collections of enhancers with a defined tissue specificity or conservation depth. These experimentally validated training sets are expected to provide a basis for a wide range of downstream computational and functional studies of enhancer function. PMID- 17130150 TI - ATTED-II: a database of co-expressed genes and cis elements for identifying co regulated gene groups in Arabidopsis. AB - Publicly available database of co-expressed gene sets would be a valuable tool for a wide variety of experimental designs, including targeting of genes for functional identification or for regulatory investigation. Here, we report the construction of an Arabidopsis thaliana trans-factor and cis-element prediction database (ATTED-II) that provides co-regulated gene relationships based on co expressed genes deduced from microarray data and the predicted cis elements. ATTED-II (http://www.atted.bio.titech.ac.jp) includes the following features: (i) lists and networks of co-expressed genes calculated from 58 publicly available experimental series, which are composed of 1388 GeneChip data in A.thaliana; (ii) prediction of cis-regulatory elements in the 200 bp region upstream of the transcription start site to predict co-regulated genes amongst the co-expressed genes; and (iii) visual representation of expression patterns for individual genes. ATTED-II can thus help researchers to clarify the function and regulation of particular genes and gene networks. PMID- 17130151 TI - The mouse Gene Expression Database (GXD): 2007 update. AB - The Gene Expression Database (GXD) provides the scientific community with an extensive and easily searchable database of gene expression information about the mouse. Its primary emphasis is on developmental studies. By integrating different types of expression data, GXD aims to provide comprehensive information about expression patterns of transcripts and proteins in wild-type and mutant mice. Integration with the other Mouse Genome Informatics (MGI) databases places the gene expression information in the context of genetic, sequence, functional and phenotypic information, enabling valuable insights into the molecular biology that underlies developmental and disease processes. In recent years the utility of GXD has been greatly enhanced by a large increase in data content, obtained from the literature and provided by researchers doing large-scale in situ and cDNA screens. In addition, we have continued to refine our query and display features to make it easier for users to interrogate the data. GXD is available through the MGI web site at http://www.informatics.jax.org/ or directly at http://www.informatics.jax.org/menus/expression_menu.shtml. PMID- 17130152 TI - Organelle DB: an updated resource of eukaryotic protein localization and function. AB - Organelle DB (http://organelledb.lsi.umich.edu) is a web-accessible relational database presenting a supplemented catalog of organelle-localized proteins and major protein complexes. Since its release in 2004, Organelle DB has grown by 20% to encompass over 30,000 proteins from 138 eukaryotic organisms. Each protein in Organelle DB is presented with its subcellular localization, primary sequence and a detailed description of its function, as available. All records in Organelle DB have been annotated using controlled vocabulary from the Gene Ontology consortium. Protein localization data are inherently visual, and Organelle DB is a significant repository of biological images, housing 1500 micrographs of yeast cells carrying stained proteins. Furthermore, we report here the development of Organelle View, an extension of Organelle DB for the interactive visualization of organelles and subcellular structures in the budding yeast Saccharomyces cerevisiae. Organelle View offers a dimensional representation of a yeast cell; users can search Organelle View for proteins of interest, and the organelles housing these proteins will be highlighted in the cell image. Among other applications, Organelle View may serve as an educational aid engaging introductory biology students through a visually 'fun' interface. Organelle View can be accessed from the Organelle DB home page or directly at http://organelleview.lsi.umich.edu. PMID- 17130153 TI - Switching base preferences of mismatch cleavage in endonuclease V: an improved method for scanning point mutations. AB - Endonuclease V (endo V) recognizes a broad range of aberrations in DNA such as deaminated bases or mismatches. It nicks DNA at the second phosphodiester bond 3' to a deaminated base or a mismatch. Endonuclease V obtained from Thermotoga maritima preferentially cleaves purine mismatches in certain sequence context. Endonuclease V has been combined with a high-fidelity DNA ligase to develop an enzymatic method for mutation scanning. A biochemical screening of site-directed mutants identified mutants in motifs III and IV that altered the base preferences in mismatch cleavage. Most profoundly, a single alanine substitution at Y80 position switched the enzyme to essentially a C-specific mismatch endonuclease, which recognized and cleaved A/C, C/A, T/C, C/T and even the previously refractory C/C mismatches. Y80A can also detect the G13D mutation in K-ras oncogene, an A/C mismatch embedded in a G/C rich sequence context that was previously inaccessible using the wild-type endo V. This investigation offers insights on base recognition and active site organization. Protein engineering in endo V may translate into better tools in mutation recognition and cancer mutation scanning. PMID- 17130154 TI - c-Fos is required for excision repair of UV-light induced DNA lesions by triggering the re-synthesis of XPF. AB - Cells deficient in c-Fos are hypersensitive to ultraviolet (UV-C) light. Here we demonstrate that mouse embryonic fibroblasts lacking c-Fos (fos-/-) are defective in the repair of UV-C induced DNA lesions. They show a decreased rate of sealing of repair-mediated DNA strand breaks and are unable to remove cyclobutane pyrimidine dimers from DNA. A search for genes responsible for the DNA repair defect revealed that upon UV-C treatment the level of xpf and xpg mRNA declined but, in contrast to the wild type (wt), did not recover in fos-/- cells. The observed decline in xpf and xpg mRNA is due to impaired re-synthesis, as shown by experiments using actinomycin D. Block of xpf transcription resulted in a lack of XPF protein after irradiation of fos-/- cells, whereas the XPF level normalized quickly in the wt. Although the xpg mRNA level was reduced, the amount of XPG protein was not altered in c-Fos-deficient cells after UV-C, due to higher stability of the XPG protein. The data suggest a new role for c-Fos in cells exposed to genotoxic stress. Being part of the transcription factor AP-1, c-Fos stimulates NER via the upregulation of xpf and thus plays a central role in the recovery of cells from UV light induced DNA damage. PMID- 17130155 TI - Fabrication and characterization of RNA aptamer microarrays for the study of protein-aptamer interactions with SPR imaging. AB - RNA microarrays were created on chemically modified gold surfaces using a novel surface ligation methodology and employed in a series of surface plasmon resonance imaging (SPRI) measurements of DNA-RNA hybridization and RNA aptamer protein binding. Various unmodified single-stranded RNA (ssRNA) oligonucleotides were ligated onto identical 5'-phosphate-terminated ssDNA microarray elements with a T4 RNA ligase surface reaction. A combination of ex situ polarization modulation FTIR measurements of the RNA monolayer and in situ SPRI measurements of DNA hybridization adsorption onto the surface were used to determine an ssRNA surface density of 4.0 x 10(12) molecules/cm2 and a surface ligation efficiency of 85 +/- 10%. The surface ligation methodology was then used to create a five component RNA microarray of potential aptamers for the protein factor IXa (fIXa). The relative surface coverages of the different aptamers were determined through a novel enzymatic method that employed SPRI measurements of a surface RNase H hydrolysis reaction. SPRI measurements were then used to correctly identify the best aptamer to fIXa, which was previously determined from SELEX measurements. A Langmuir adsorption coefficient of 1.6 x 10(7) M(-1) was determined for fIXa adsorption to this aptamer. Single-base variations from this sequence were shown to completely destroy the aptamer-fIXa binding interaction. PMID- 17130156 TI - Use of thermolytic protective groups to prevent G-tetrad formation in CpG ODN type D: structural studies and immunomodulatory activity in primates. AB - CpG oligodeoxynucleotides (ODN) show promise as immunoprotective agents and vaccine adjuvants. CpG ODN type D were shown to improve clinical outcome in rhesus macaques challenged with Leishmania major. These ODN have a self complementary core sequence and a 3' end poly(G) track that favors G-tetrad formation leading to multimerization. Although multimerization appears necessary for localization to early endosomes and signaling via Toll-like receptor 9 (TLR 9), it can result in product polymorphisms, aggregation and precipitation, thereby hampering their clinical applications. This study shows that functionalizing the poly(G) track of D ODN with thermolytic 2-(N-formyl-N methyl)aminoethyl (fma) phosphate/thiophosphate protecting groups (pro-D ODN) reduces G-tetrad formation in solution, while allowing tetrad formation inside the cell where the potassium concentration is higher. Temperature-dependent cleavage of the fma groups over time further promoted formation of stable G tetrads. Peripheral blood cells internalized pro-D ODN efficiently, inducing high levels of IFNalpha, IL-6, IFNgamma and IP-10 and triggering dendritic cell maturation. Administration of pro-D35 to macaques challenged with L.major significantly increased the number of antigen-specific IFNgamma-secreting PBMC and reduced the severity of the skin lesions demonstrating immunoprotective activity of pro-D ODN in vivo. This technology fosters the development of more efficient immunotherapeutic oligonucleotide formulations for the treatment of allergies, cancer and infectious diseases. PMID- 17130157 TI - Transcriptional regulation of the cyclin-dependent kinase inhibitor 1A (p21) gene by NFI in proliferating human cells. AB - The cyclin-dependent kinase inhibitor 1A (CDKN1A), also known as p21 (WAF1/CIP1) modulates cell cycle, apoptosis, senescence and differentiation via specific protein-protein interactions with the cyclins, cyclin-dependent kinase (Cdk), and many others. Expression of the p21 gene is mainly regulated at the transcriptional level. By conducting both ligation-mediated PCR (LMPCR) and chromatin immunoprecipitation (ChIP) in vivo, we identified a functional target site for the transcription factor, nuclear factor I (NFI), in the basal promoter from the p21 gene. Transfection of recombinant constructs bearing mutations in the p21 NFI site demonstrated that NFI acts as a repressor of p21 gene expression in various types of cultured cells. Inhibition of NFI in human skin fibroblasts through RNAi considerably increased p21 promoter activity suggesting that NFI is a key repressor of p21 transcription. Over-expression of each of the four NFI isoforms in HCT116 cells established that each of them contribute to various extend to the repression of the p21 gene. Most of all, over-expression of NFI-B in doxorubicin, growth-arrested HCT116 increased the proportion of cells in the S phase of the cell cycle whereas NFI-A and NFI-X reduced it, thereby establishing a role for NFI in the cell cycle dependent expression of p21. PMID- 17130158 TI - CoMoDis: composite motif discovery in mammalian genomes. AB - Specificity of mammalian gene regulatory regions is achieved to a large extent through the combinatorial binding of sets of transcription factors to distinct binding sites, discrete combinations of which are often referred to as regulatory modules. Identification and subsequent characterization of gene regulatory modules will be a key step in assembling transcriptional regulatory networks from gene expression profiling data, with the ultimate goal of unravelling the regulatory codes that govern gene expression in various cell types. Here we describe the new bioinformatics tool, Composite Motif Discovery (CoMoDis), which streamlines computational identification of novel regulatory modules starting from a single seed motif. Seed motifs represent binding sites conserved across mammalian species. CoMoDis facilitates novel motif discovery by automating the extraction of DNA sequences flanking seed motifs and streamlining downstream motif discovery using a variety of tools, including several that utilize phylogenetic conservation criteria. CoMoDis is available at http://hscl.cimr.cam.ac.uk/CoMoDis_portal.html. PMID- 17130159 TI - Self-splicing of a group IIC intron: 5' exon recognition and alternative 5' splicing events implicate the stem-loop motif of a transcriptional terminator. AB - Bacterial IIC introns are a newly recognized subclass of group II introns whose ribozyme properties have not been characterized in detail. IIC introns are typically located downstream of transcriptional terminator motifs (inverted repeat followed by T's) or other inverted repeats in bacterial genomes. Here we have characterized the self-splicing activity of a IIC intron, B.h.I1, from Bacillus halodurans. B.h.I1 self-splices in vitro through hydrolysis to produce linear intron, but interestingly, additional unexpected products were formed that were highly dependent on ionic conditions. These products were determined to represent alternative splicing events at the 5' junction and cleavages throughout the RNA transcript. The alternative splicing and cleavage events occurred at cryptic splice sites containing stem-loop and IBS1 motifs, suggesting that the 5' exon is recognized by both elements. These results provide the first example of a group II intron that uses 5' splice sites nonadjacent to the ribozyme structure. Furthermore, the data suggest that IIC introns differ from IIA and IIB introns with respect to 5' exon definition, and that the terminator stem-loop substitutes in part for the missing IBS2-EBS2 (intron and exon binding sites 2) interaction. PMID- 17130160 TI - Amino acid residue doublet propensity in the protein-RNA interface and its application to RNA interface prediction. AB - Protein-RNA interactions play essential roles in a number of regulatory mechanisms for gene expression such as RNA splicing, transport, translation and post-transcriptional control. As the number of available protein-RNA complex 3D structures has increased, it is now possible to statistically examine protein-RNA interactions based on 3D structures. We performed computational analyses of 86 representative protein-RNA complexes retrieved from the Protein Data Bank. Interface residue propensity, a measure of the relative importance of different amino acid residues in the RNA interface, was calculated for each amino acid residue type (residue singlet interface propensity). In addition to the residue singlet propensity, we introduce a new residue-based propensity, which gives a measure of residue pairing preferences in the RNA interface of a protein (residue doublet interface propensity). The residue doublet interface propensity contains much more information than the sum of two singlet propensities alone. The prediction of the RNA interface using the two types of propensities plus a position-specific multiple sequence profile can achieve a specificity of about 80%. The prediction method was then applied to the 3D structure of two mRNA export factors, TAP (Mex67) and UAP56 (Sub2). The prediction enables us to point out candidate RNA interfaces, part of which are consistent with previous experimental studies and may contribute to elucidation of atomic mechanisms of mRNA export. PMID- 17130161 TI - Rapid detection of similarity in protein structure and function through contact metric distances. AB - The characterization of biological function among newly determined protein structures is a central challenge in structural genomics. One class of computational solutions to this problem is based on the similarity of protein structure. Here, we implement a simple yet efficient measure of protein structure similarity, the contact metric. Even though its computation avoids structural alignments and is therefore nearly instantaneous, we find that small values correlate with geometrical root mean square deviations obtained from structural alignments. To test whether the contact metric detects functional similarity, as defined by Gene Ontology (GO) terms, it was compared in large-scale computational experiments to four other measures of structural similarity, including alignment algorithms as well as alignment independent approaches. The contact metric was the fastest method and its sensitivity, at any given specificity level, was a close second only to Fast Alignment and Search Tool--a structural alignment method that is slower by three orders of magnitude. Critically, nearly 40% of correct functional inferences by the contact metric were not identified by any other approach, which shows that the contact metric is complementary and computationally efficient in detecting functional relationships between proteins. A public 'Contact Metric Internet Server' is provided. PMID- 17130162 TI - Absolute enrichment: gene set enrichment analysis for homeostatic systems. AB - The Gene Set Enrichment Analysis (GSEA) identifies sets of genes that are differentially regulated in one direction. Many homeostatic systems will include one limb that is upregulated in response to a downregulation of another limb and vice versa. Such patterns are poorly captured by the standard formulation of GSEA. We describe a technique to identify groups of genes (which sometimes can be pathways) that include both up- and down-regulated components. This approach lends insights into the feedback mechanisms that may operate, especially when integrated with protein interaction databases. PMID- 17130163 TI - Template properties of mutagenic cytosine analogues in reverse transcription. AB - We have studied the mutagenic properties of ribonucleotide analogues by reverse transcription to understand their potential as antiretroviral agents by mutagenesis of the viral genome. The templating properties of nucleotide analogues including 6-(beta-D-ribofuranosyl)-3,4-dihydro-8H-pyrimido[4,5 c](1,2)oxazin-7-one, N4-hydroxycytidine, N4-methoxycytidine, N4-methylcytidine and 4-semicarbazidocytidine, which have been reported to exhibit ambiguous base pairing properties, were examined. We have synthesized RNA templates using T3 RNA polymerase, and investigated the specificity of the incorporation of deoxyribonucleoside triphosphates opposite these cytidine analogues in RNA by HIV and AMV reverse transcriptases. Except for N4-methylcytidine, both enzymes incorporated both dAMP and dGMP opposite these analogues in RNA. This indicates that they would be highly mutagenic if present in viral RNA. To study the basis of the differences among the analogues in the incorporation ratios of dAMP to dGMP, we have carried out kinetic analysis of incorporation opposite the analogues at a defined position in RNA templates. In addition, we examined whether the triphosphates of these analogues were incorporated competitively into RNA by human RNA polymerase II. Our present data supports the view that these cytidine analogues are mutagenic when incorporated into RNA, and that they may therefore be considered as candidates for antiviral agents by causing mutations to the retroviral genome. PMID- 17130164 TI - Sequence-dependent gating of an ion channel by DNA hairpin molecules. AB - DNA hairpins produce ionic current signatures when captured by the alpha hemolysin nano-scale pore under conditions of single molecule electrophoresis. Gating patterns produced by individual DNA hairpins when captured can be used to distinguish differences of a single base pair or even a single nucleotide [Vercoutere,W.A. et al. (2003) Nucleic Acids Res., 31, 1311-1318]. Here we investigate the mechanism(s) that may account for the ionic current gating signatures. The ionic current resistance profile of conductance states produced by DNA hairpin molecules with 3-12 bp stems showed a plateau in resistance between 10 and 12 bp, suggesting that hairpins with 10-12 bp stems span the pore vestibule. DNA hairpins with 9-12 bp stems produced gating signatures with the same relative conductance states. Systematic comparison of the conductance state dwell times and apparent activation energies for a series of 9-10 bp DNA hairpins suggest that the 3' and 5' ends interact at or near the limiting aperture within the vestibule of the alpha-hemolysin pore. The model presented may be useful in predicting and interpreting DNA detection using nanopore detectors. In addition, this well-defined molecular system may prove useful for investigating models of ligand-gated channels in biological membranes. PMID- 17130165 TI - Development of a novel site-specific mutagenesis assay using MALDI-ToF MS (SSMA MS). AB - We have developed and validated a novel site-specific mutagenesis assay, termed SSMA-MS, which incorporates MALDI-ToF mass spectrometry (MALDI-MS) analysis as a means of determining the mutations induced by a single DNA adduct. The assay involves ligating an adducted deoxyoligonucleotide into supF containing pSP189 plasmid. The plasmid is transfected into human Ad293 kidney cells allowing replication and therefore repair or a mutagenic event to occur. Escherichia coli indicator bacteria are transformed with recovered plasmid and plasmids containing the insert are identified colormetrically, as they behave as frameshift mutations. The plasmid is then amplified and digested using a restriction cocktail of Mbo11 and Mnl1 to yield 12 bp deoxyoligonucleotides, which are characterized by MALDI-MS. MALDI-MS takes advantage of the difference in molecular weight between bases to identify any induced mutations. This analysis method therefore provides qualitative and quantitative information regarding the type and frequency of mutations induced. This assay was developed and validated using an O(6)-methyl-2'-deoxyguanosine adduct, which induced the expected GC-->AT substitutions, when replicated in human or bacterial cells. This approach can be applied to the study of any DNA adduct in any biologically relevant gene sequence (e.g. p53) in human cells and would be particularly amenable to high-throughput analysis. PMID- 17130166 TI - Molecular dysfunction associated with the human mitochondrial 3302A>G mutation in the MTTL1 (mt-tRNALeu(UUR)) gene. AB - The gene encoding mt-tRNA(Leu(UUR)), MT-TL1, is a hotspot for pathogenic mtDNA mutations. Amongst the first to be described was the 3302A>G transition which resulted in a substantial accumulation in patient muscle of RNA19, an unprocessed RNA intermediate including mt-16S rRNA, mt-tRNA(Leu(UUR)) and MTND1. We have now been able to further assess the molecular aetiology associated with 3302A>G in transmitochondrial cybrids. Increased steady-state levels of RNA19 was confirmed, although not to the levels previously reported in muscle. This data was consistent with an increase in RNA19 stability. The mutation resulted in decreased mt-tRNA(Leu(UUR)) levels, but its stability was unchanged, consistent with a defect in RNA19 processing responsible for low tRNA levels. A partial defect in aminoacylation was also identified, potentially caused by an alteration in tRNA structure. These deficiencies lead to a severe defect in respiration in the transmitochondrial cybrids, consistent with the profound mitochondrial disorder originally associated with this mutation. PMID- 17130167 TI - Evidence of the neuron-restrictive silencer factor (NRSF) interaction with Sp3 and its synergic repression to the mu opioid receptor (MOR) gene. AB - Previously, we reported that the neuron-restrictive silencer element (NRSE) of mu opioid receptor (MOR) functions as a critical regulator to repress the MOR transcription in specific neuronal cells, depending on neuron-restriction silence factor (NRSF) expression levels [C.S.Kim, C.K.Hwang, H.S.Choi, K.Y.Song, P.Y.Law, L.N. Wei and H.H.Loh (2004) J. Biol. Chem., 279, 46464-46473]. Herein, we identify a conserved GC sequence next to NRSE region in the mouse MOR gene. The inhibition of Sp family factors binding to this GC box by mithramycin A led to a significant increase in the endogenous MOR transcription. In the co immunoprecipitation experiment, NRSF interacted with the full-length Sp3 factor, but not with Sp1 or two short Sp3 isoforms. The sequence specific and functional binding by Sp3 at this GC box was confirmed by in vitro gel-shift assays using either in vitro translated proteins or nuclear extract, and by in vivo chromatin immunoprecipitation assays. Transient transfection assays showed that Sp3-binding site of the MOR gene is a functionally synergic repressor element with NRSE in NS20Y cells, but not in the NRSF negative PC12 cells. The results suggest that the synergic interaction between NRSF and Sp3 is required to negatively regulate MOR gene transcription and that transcription of MOR gene would be governed by the context of available transcription factors rather than by a master regulator. PMID- 17130168 TI - A combinatorial approach to create artificial homing endonucleases cleaving chosen sequences. AB - Meganucleases, or homing endonucleases (HEs) are sequence-specific endonucleases with large (>14 bp) cleavage sites that can be used to induce efficient homologous gene targeting in cultured cells and plants. These findings have opened novel perspectives for genome engineering in a wide range of fields, including gene therapy. However, the number of identified HEs does not match the diversity of genomic sequences, and the probability of finding a homing site in a chosen gene is extremely low. Therefore, the design of artificial endonucleases with chosen specificities is under intense investigation. In this report, we describe the first artificial HEs whose specificity has been entirely redesigned to cleave a naturally occurring sequence. First, hundreds of novel endonucleases with locally altered substrate specificity were derived from I-CreI, a Chlamydomonas reinhardti protein belonging to the LAGLIDADG family of HEs. Second, distinct DNA-binding subdomains were identified within the protein. Third, we used these findings to assemble four sets of mutations into heterodimeric endonucleases cleaving a model target or a sequence from the human RAG1 gene. These results demonstrate that the plasticity of LAGLIDADG endonucleases allows extensive engineering, and provide a general method to create novel endonucleases with tailored specificities. PMID- 17130169 TI - Identification of degenerate motifs using position restricted selection and hybrid ranking combination. AB - The identification of regulatory elements recognized by transcription factors and chromatin remodeling factors is essential to studying the regulation of gene expression. When no auxiliary data, such as orthologous sequences or expression profiles, are used, the accuracy of most tools for motif discovery is strongly influenced by the motif degeneracy and the lengths of sequence. Since suitable auxiliary data may not always be available, more work must be conducted to enhance tool performance to identify transcription elements in the metazoan. A non-alignment-based algorithm, MotifSeeker, is proposed to enhance the accuracy of discovering degenerate motifs. MotifSeeker utilizes the property that variable sites of transcription elements are usually position-specific to reduce exposure to noise. Consequently, the efficiency and accuracy of motif identification are improved. Using data fusion, the ranking process integrates two measures of motif significance, resulting in a more robust significance measure. Testing results for the synthetic data reveal that the accuracy of MotifSeeker is less sensitive to the motif degeneracy and the length of input sequences. Furthermore, MotifSeeker has been tested on a well-known benchmark [M. Tompa, N. Li, T.L. Bailey, G.M. Church, B. De Moor, E. Eskin, A.V. Favorov, M.C. Frith, Y. Fu, W.J. Kent, et al. (2005) Nat. Biotechnol., 23, 137-144], yielding a correlation coefficient of 0.262, which compares favorably with those of other tools. The high applicability of MotifSeeker to biological data is further demonstrated experimentally on regulons of Saccharomyces cerevisiae and liver-specific genes with experimentally verified regulatory elements. PMID- 17130170 TI - Sequence-dependent formation of intrastrand crosslink products from the UVB irradiation of duplex DNA containing a 5-bromo-2'-deoxyuridine or 5-bromo-2' deoxycytidine. AB - The replacement of thymidine with 5-bromo-2'-deoxyuridine (BrdU) is well-known to sensitize cells to ionizing radiation and photoirradiation. We reported here the sequence-dependent formation of intrastrand crosslink products from the UVB irradiation of duplex oligodeoxynucleotides harboring a BrdU or its closely related 5-bromo-2'-deoxycytidine (BrdC). Our results showed that two types of crosslink products could be induced from d(BrCG), d(BrUG), d(GBrU), or d(ABrU); the C(5) of cytosine or uracil could be covalently bonded to the N(2) or C(8) of its neighboring guanine, and the C(5) of uracil could couple with the C(2) or C(8) of its neighboring adenine. By using those crosslink product-bearing dinucleoside monophosphates as standards, we demonstrated, by using liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS), that all the crosslink products described above except d(G[N(2)-5]U) and d(G[N(2)-5]C) could form in duplex DNA. In addition, LC-MS/MS quantification results revealed that both the nature of the halogenated pyrimidine base and its 5' flanking nucleoside affected markedly the generation of intrastrand crosslink products. The yields of crosslink products were much higher while the 5' neighboring nucleoside was a dG than while it was a dA, and BrdC induced the formation of crosslink products much more efficiently than BrdU. The formation of intrastrand crosslink products from these halopyrimidines in duplex DNA may account for the photosensitizing effects of these nucleosides. PMID- 17130171 TI - Activation of RegB endoribonuclease by S1 ribosomal protein requires an 11 nt conserved sequence. AB - The T4 RegB endoribonuclease cleaves specifically in the middle of the -GGAG- sequence, leading to inactivation and degradation of early phage mRNAs. In vitro, RegB activity is very weak but can be enhanced 10- to 100-fold by the Escherichia coli ribosomal protein S1. Not all RNAs carrying the GGAG motif are cleaved by RegB, suggesting that additional information is required to obtain a complete RegB target site. In this work, we find that in the presence of S1, the RegB target site is an 11 nt long single-stranded RNA carrying the 100% conserved GGA triplet at the 5' end and a degenerate, A-rich, consensus sequence immediately downstream. Our data support the notion that RegB alone recognizes only the trinucleotide GGA, which it cleaves very inefficiently, and that stimulation of RegB activity by S1 depends on the nucleotide immediately 3' to -GGA-. PMID- 17130172 TI - Sequence comparison by sequence harmony identifies subtype-specific functional sites. AB - Multiple sequence alignments are often used to reveal functionally important residues within a protein family. They can be particularly useful for the identification of key residues that determine functional differences between protein subfamilies. We present a new entropy-based method, Sequence Harmony (SH) that accurately detects subfamily-specific positions from a multiple sequence alignment. The SH algorithm implements a novel formula, able to score compositional differences between subfamilies, without imposing conservation, in a simple manner on an intuitive scale. We compare our method with the most important published methods, i.e. AMAS, TreeDet and SDP-pred, using three well studied protein families: the receptor-binding domain (MH2) of the Smad family of transcription factors, the Ras-superfamily of small GTPases and the MIP-family of integral membrane transporters. We demonstrate that SH accurately selects known functional sites with higher coverage than the other methods for these test cases. This shows that compositional differences between protein subfamilies provide sufficient basis for identification of functional sites. In addition, SH selects a number of sites of unknown function that could be interesting candidates for further experimental investigation. PMID- 17130173 TI - The natural history of the WRKY-GCM1 zinc fingers and the relationship between transcription factors and transposons. AB - WRKY and GCM1 are metal chelating DNA-binding domains (DBD) which share a four stranded fold. Using sensitive sequence searches, we show that this WRKY-GCM1 fold is also shared by the FLYWCH Zn-finger domain and the DBDs of two classes of Mutator-like element (MULE) transposases. We present evidence that they share a stabilizing core, which suggests a possible origin from a BED finger-like intermediate that was in turn ultimately derived from a C2H2 Zn-finger domain. Through a systematic study of the phyletic pattern, we show that this WRKY-GCM1 superfamily is a widespread eukaryote-specific group of transcription factors (TFs). We identified several new members across diverse eukaryotic lineages, including potential TFs in animals, fungi and Entamoeba. By integrating sequence, structure, gene expression and transcriptional network data, we present evidence that at least two major global regulators belonging to this superfamily in Saccharomyces cerevisiae (Rcs1p and Aft2p) have evolved from transposons, and attained the status of transcription regulatory hubs in recent course of ascomycete yeast evolution. In plants, we show that the lineage-specific expansion of WRKY-GCM1 domain proteins acquired functional diversity mainly through expression divergence rather than by protein sequence divergence. We also use the WRKY-GCM1 superfamily as an example to illustrate the importance of transposons in the emergence of new TFs in different lineages. PMID- 17130174 TI - The Agrobacterium VirE3 effector protein: a potential plant transcriptional activator. AB - During the infection of plants, Agrobacterium tumefaciens introduces several Virulence proteins including VirE2, VirF, VirD5 and VirE3 into plant cells in addition to the T-DNA. Here, we report that double mutation of virF and virE3 leads to strongly diminished tumor formation on tobacco, tomato and sunflower. The VirE3 protein is translated from a polycistronic mRNA containing the virE1, virE2 and virE3 genes, in Agrobacterium. The VirE3 protein has nuclear localization sequences, which suggests that it is transported into the plant cell nucleus upon translocation. Indeed we show here that VirE3 interacts in vitro with importin-alpha and that a VirE3-GFP fusion protein is localized in the nucleus. VirE3 also interacts with two other proteins, viz. pCsn5, a component of the COP9 signalosome and pBrp, a plant specific general transcription factor belonging to the TFIIB family. We found that VirE3 is able to induce transcription in yeast when bound to DNA through the GAL4-BD. Our data indicate that the translocated effector protein VirE3 is transported into the nucleus and there it may interact with the transcription factor pBrp to induce the expression of genes needed for tumor development. PMID- 17130175 TI - Defining and investigating occupational asthma: a consensus approach. AB - BACKGROUND: At present there is no internationally agreed definition of occupational asthma and there is a lack of guidance regarding the resources that should be readily available to physicians running specialist occupational asthma services. AIMS: To agree a working definition of occupational asthma and to develop a framework of resources necessary to run a specialist occupational asthma clinic. METHOD: A modified RAND appropriateness method was used to gain a consensus of opinion from an expert panel of clinicians running specialist occupational asthma clinics in the UK. RESULTS: Consensus was reached over 10 terms defining occupational asthma including: occupational asthma is defined as asthma induced by exposure in the working environment to airborne dusts vapours or fumes, with or without pre-existing asthma; occupational asthma encompasses the terms "sensitiser-induced asthma" and "acute irritant-induced asthma" (reactive airways dysfunction syndrome (RADS)); acute irritant-induced asthma is a type of occupational asthma where there is no latency and no immunological sensitisation and should only be used when a single high exposure has occurred; and the term "work-related asthma" can be used to include occupational asthma, acute irritant-induced asthma (RADS) and aggravation of pre-existing asthma. Disagreement arose on whether low dose irritant-induced asthma existed, but the panel agreed that if it did exist they would include it in the definition of "work-related asthma". The panel agreed on a set of 18 resources which should be available to a specialist occupational asthma service. These included pre bronchodilator FEV1 and FVC (% predicted); peak flow monitoring (and plotting of results, OASYS II analysis); non-specific provocation challenge in the laboratory and specific IgE to a wide variety of occupational agents. CONCLUSION: It is hoped that the outcome of this process will improve uniformity of definition and investigation of occupational asthma across the UK. PMID- 17130176 TI - Oxidative DNA damage induced by Ochratoxin A in the HK-2 human kidney cell line: evidence of the relationship with cytotoxicity. AB - Ochratoxin A (OTA) is a mycotoxin produced by species of the genera Aspergillus and Penicillium. The kidneys are the target organ of this mycotoxin and it is considered a potent renal carcinogen in male rats. The mechanisms of its genotoxicity and carcinogenicity have been studied thoroughly, but controversial results have been published. The aim of this study was to evaluate the ability of OTA to produce single-strand DNA breaks and oxidative DNA damage in the human renal proximal tubular epithelial cell line (HK-2), due to the fact that there is no study on human kidney cells as the toxic target. In addition, we attempted to determine if biotransformation processes mediate OTA genotoxicity. Therefore, single-cell gel electrophoresis assay (comet assay) was performed after 3h- and 6h-treatments using different OTA concentrations, both cytotoxic and non cytotoxic, in order to be able to distinguish a genotoxic effect of the mycotoxin from an indirect effect derived from its general cellular toxicity. No effect was shown where no cytotoxicity was found, both in the presence and in the absence of metabolic activation (10% rat liver S9-mix). However, oxidative DNA damage was shown at cytotoxic concentrations when formamidopyrimidine DNA glycosylase (FPG) and endonucleaseIII (EndoIII) were introduced in the assay with or without metabolic activation. Furthermore, at these concentrations, an elevation of reactive oxygen species was measured and pre-incubation with N-acetyl-L-cysteine was able to produce a slight protective effect on OTA-induced oxidative DNA damage as well as cytotoxicity. These data suggest that OTA is not acting as a direct genotoxic carcinogen and that oxidative stress is implicated in the genotoxicity and cytotoxicity observed in these human renal cells. PMID- 17130177 TI - Microchip capillary electrophoresis for point-of-care analysis of lithium. AB - BACKGROUND: Microchip capillary electrophoresis (CE) is a promising method for chemical analysis of complex samples such as whole blood. We evaluated the method for point-of-care testing of lithium. METHODS: Chemical separation was performed on standard glass microchip CE devices with a conductivity detector as described in previous work. Here we demonstrate a new sample-to-chip interface. Initially, we took a glass capillary as a sample collector for whole blood from a finger stick. In addition, we designed a novel disposable sample collector and tested it against the clinical standard at the hospital (Medisch Spectrum Twente). Both types of collectors require < 10 microL of test fluid. The collectors contain an integrated filter membrane, which prevents the transfer of blood cells into the microchip. The combination of such a sample collector with microchip CE allows point-of-care measurements without the need for off-chip sample treatment. This new on-chip protocol was verified against routine lithium testing of 5 patients in the hospital. RESULTS: Sodium, lithium, magnesium, and calcium were separated in < 20 s. The detection limit for lithium was 0.15 mmol/L. CONCLUSIONS: The new microchip CE system provides a convenient and rapid method for point-of-care testing of electrolytes in serum and whole blood. PMID- 17130178 TI - Potential utility of plasma fatty acid analysis in the diagnosis of cystic fibrosis. AB - BACKGROUND: An altered distribution of fatty acids in cells and tissues is found in patients with cystic fibrosis (CF). In this study, we assessed the potential role of plasma fatty acid analysis in the diagnosis of CF. METHODS: In this 2 part study, we first used gas chromatography-mass spectrometry to analyze fatty acids in plasma from 13 CF patients and 11 controls without CF. We then used the fatty acid distribution data to identify the fatty acids or multiple fatty acid calculations most effective in identifying CF patients. Part 2 of the study was a blinded analysis of 10 CF patients and 9 controls to directly test the effectiveness of the diagnostic parameters for CF identified from the plasma fatty acid analysis. RESULTS: In the nonblinded trial, the multiplication product of (18:2 n-6) x (22:6 n-3) (each as percentage of total plasma fatty acid) was the most effective indicator for distinguishing patients with CF from controls (P = 0.0003). In part 2 (the blinded trial), this multiplication product was also the most effective indicator for distinguishing CF patients from controls (P = 0.0008). CONCLUSIONS: The product of (18:2 n-6) x (22:6 n-3) is effective for distinguishing CF patients from persons without CF. This diagnostic marker may have value as an alternative to the sweat chloride test in selected patients being evaluated for CF. PMID- 17130179 TI - Changes in biochemical markers after lower limb fractures. AB - BACKGROUND: The bone remodeling sequence after bone fracture changes the concentrations of biochemical bone markers, but the relationships of fracture size and of healing time to changes in biomarkers are unclear. The present pilot study was undertaken to determine the changes found in serum bone markers after plate osteosynthesis of closed distal tibial and malleolar fractures during a study period of 24 weeks. METHODS: We measured tatrate-resistant acid phosphatase (TRACP 5b), collagen type I C-terminal telopeptide (ICTP), bone-specific alkaline phosphatase (bone ALP), osteocalcin (OC), procollagen type I C-terminal propeptide (PICP), procollagen type III N-terminal propeptide (PIIINP), and human cartilage glycoprotein 39 (YKL-40) in 20 patients with lower limb fractures (10 malleolar, 10 tibia). A physical examination and radiographs were completed to assess evidence of union. RESULTS: All malleolar fractures healed within 6 weeks, whereas 2 tibial fractures did not show complete bone healing after 24 weeks. Changes were comparable but more pronounced in the tibia group, and marker concentrations remained increased at the end of study (bone ALP, 86 vs 74 U/L; OC, 14.9 vs 7.7 microg/L; ICTP: 5.6 vs 3.3 microg/L at day 84 after osteosynthesis, P <0.05 in tibia; 80 vs 70 U/L, 8 vs 5.2 microg/L, and 3.5 vs 3.2 microg/L, respectively, in the malleolar fracture group). CONCLUSIONS: In normal bone healing, changes in bone turnover markers were primarily dependent on the fracture size. Delayed tibia fracture healing may involve a disturbance in bone remodeling. PMID- 17130180 TI - Candidate gene genotypes, along with conventional risk factor assessment, improve estimation of coronary heart disease risk in healthy UK men. AB - BACKGROUND: One of the aims of cardiovascular genetics is to test the efficacy of the use of genetic information to predict cardiovascular risk. We therefore investigated whether inclusion of a set of common variants in candidate genes along with conventional risk factor (CRF) assessment enhanced coronary heart disease (CHD)-risk algorithms. METHODS: We followed middle-aged men in the prospective Northwick Park Heart Study II (NPHSII) for 10.8 years and analyzed complete trait and genotype information available on 2057 men (183 CHD events). RESULTS: Of the 12 genes previously associated with CHD risk, in stepwise multivariate risk analysis, uncoupling protein 2 (UCP2; P = 0.0001), apolipoprotein E (APOE; P = 0.0003), lipoprotein lipase (LPL; P = 0.007), and apolipoprotein AIV (APOA4; P = 0.04) remained in the model. Their combined area under the ROC curve (A(ROC)) was 0.62 (0.58-0.66) [12.6% detection rate for a 5% false positive rate (DR(5))]. The A(ROC) for the CRFs age, triglyceride, cholesterol, systolic blood pressure, and smoking was 0.66 (0.61-0.70) (DR(5) = 14.2%). Combining CRFs and genotypes significantly improved discrimination (P = 0.001). Inclusion of previously demonstrated interactions of smoking with LPL, interleukin-6 (IL6), and platelet/endothelial cell adhesion molecule (PECAM1) genotypes increased the A(ROC) to 0.72 (0.68-0.76) for a DR(5) of 19.1% (P = 0.01 vs CRF combined with genotypes). CONCLUSIONS: For a modest panel of selected genotypes, CHD-risk estimates incorporating CRFs and genotype-risk factor interactions were more effective than risk estimates that used CRFs alone. PMID- 17130181 TI - Real-time RT-PCR quantification of human telomerase reverse transcriptase splice variants in tumor cell lines and non-small cell lung cancer. AB - BACKGROUND: We developed and validated a real-time reverse transcription (RT)-PCR for the quantification of 4 individual human telomerase reverse transcriptase (TERT) splice variants (alpha+beta+, alpha-beta+, alpha+beta-, alpha-beta-) in tumor cell lines and non-small cell lung cancer (NSCLC). METHODS: We used in silico designed primers and a common TaqMan probe for highly specific amplification of each TERT splice variant, PCR transcript-specific DNA external standards as calibrators, and the MCF-7 cell line for the development and validation of the method. We then quantified TERT splice variants in 6 tumor cell lines and telomerase activity and TERT splice variant expression in cancerous and paired noncancerous tissue samples from 28 NSCLC patients. RESULTS: In most tumor cell lines, we observed little variation in the proportion of TERT splice variants. The alpha+beta- splice variant showed the highest expression and alpha beta+ and alpha-beta- the lowest. Quantification of the 4 TERT splice variants in NSCLC and surrounding nonneoplastic tissues showed the highest expression percentage for the alpha+beta- variant in both NSCLC and adjacent nonneoplastic tissue samples, followed by alpha+beta+, with the alpha-beta+ and alpha-beta- splice variants having the lowest expression. In the NSCLC tumors, the alpha+beta+ variant had higher expression than other splice variants, and its expression correlated with telomerase activity, overall survival, and disease free survival. CONCLUSIONS: Real-time RT-PCR quantification is a specific, sensitive, and rapid method that can elucidate the biological role of TERT splice variants in tumor development and progression. Our results suggest that the expression of the TERT alpha+beta+ splice variant may be an independent negative prognostic factor for NSCLC patients. PMID- 17130182 TI - A systematic review on the efficacy of cytoreductive surgery combined with perioperative intraperitoneal chemotherapy for diffuse malignancy peritoneal mesothelioma. AB - BACKGROUND: In the past, diffuse malignant peritoneal mesothelioma (DMPM) was regarded as a preterminal condition. The length of survival was dependent upon the aggressive versus indolent biologic behavior of the neoplasm. The overall median survival was approximately 1 year after systemic chemotherapy. Cytoreductive surgery (CRS) combined with perioperative intraperitoneal chemotherapy (PIC) has been used as a treatment alternative, but the efficacy of this combined treatment remains to be established. PATIENTS AND METHODS: Searches for relevant studies published in peer-reviewed medical journals on CRS and PIC for DMPM before May 2006 were carried out on six databases. The reference lists of all retrieved articles were reviewed for further identification of potentially relevant studies. Expert academic surgeons in Washington, DC, USA were asked whether they knew about any important unpublished data. Two investigators independently evaluated each study according to predefined criteria. The quality of each study was assessed. Clinical effectiveness was synthesized through a narrative review with full tabulation of results of all included studies. RESULTS: Seven prospective observational studies from six tertiary institutions were available, allowing 240 DMPM patients for assessment. The median survival ranged from 34-92 months. The 1-, 3- and 5-year survival varied from 60% to 88%, 43% to 65% and 29% to 59%, respectively. The perioperative morbidity varied from 25% to 40% and mortality ranged from 0% to 8%. CONCLUSIONS: This systematic review evaluated the current evidence for CRS and PIC for DMPM. Seven observational studies were available for assessment, which demonstrated an improved overall survival, as compared to historical controls, using systemic chemotherapy and palliative surgery. PMID- 17130183 TI - The superantigen-induced polarization of T cells in rat peripheral lymph nodes is influenced by genetic polymorphisms in the IL-4 and IL-6 gene clusters. AB - In recent years, it has become clear that the polarization of T cells depends on the genetic background. However, due to the complexity of the genetic background of each animal, a direct comparison of the phenotype is difficult. In this study, a new rat strain LEW.BN-4-10 carrying the chromosomal regions on chromosomes 4 and 10, which harbor IL-6 and IL-4 gene clusters of BN, has been bred on the genetic background of LEW. It was asked whether these two gene clusters influence the polarization of T cell responses. As a model, the Mycoplasma arthritidis mitogen (MAM)-induced inflammation was used focusing on the microenvironment of the draining lymph node (LN). The effect of differences in these regions was tested by comparing LEW.BN-4-10 and LEW rats under steady-state conditions and upon injection of MAM into the forepaw. Under steady-state conditions, the two strains showed differences in the dendritic cell (DC) subset composition. When MAM was injected, the number of T cells in LEW.BN-4-10 rats producing T(h)2 cytokines such as IL-4 and IL-13 was significantly increased compared with LEW. The data suggest that these differences in the microenvironments in LN of LEW and LEW.BN-4-10 rats resulted in different susceptibility to the disease (increase of cells in LN and paw swelling). In addition, deviations in the distribution and function of injected effector T cells were found in the LN of LEW and LEW.BN-4-10 rats after MAM treatment. The data indicate that the IL-6 and IL-4 gene clusters are involved in polarizing T cell responses in vivo. PMID- 17130185 TI - Are source and sink strengths genetically linked in maize plants subjected to water deficit? A QTL study of the responses of leaf growth and of Anthesis Silking Interval to water deficit. AB - Leaf growth and Anthesis-Silking Interval (ASI) are the main determinants of source and sink strengths of maize via their relations with light interception and yield, respectively. They depend on the abilities of leaves and silks to expand under fluctuating environmental conditions, so the possibility is raised that they may have a partly common genetic determinism. This possibility was tested in a mapping population which segregates for ASI. Maximum leaf elongation rate per unit thermal time (parameter a) and the slopes of its responses to evaporative demand and soil water status (parameters b and c) were measured in greenhouse and growth chamber experiments, in two series of 120 recombinant inbred lines (RILs) studied in 2004 and 2005 with 33 RILs in common both years. ASI was measured in three and five fields under well-watered conditions and water deficit, respectively. For each RIL, the maximum elongation rate per unit thermal time was reproducible over several experiments in well-watered plants. It was accounted for by five QTLs, among which three co-localized with QTLs of ASI of well-watered plants. The alleles conferring high leaf elongation rate conferred a low ASI (high silk elongation rate). The responses of leaf elongation rate to evaporative demand and to predawn leaf water potential were linear, allowing each RIL to be characterized by the slopes of these response curves. These slopes had three QTLs in common with ASI of plants under water deficit. The allele for leaf growth maintenance was, in all cases, that for shorter ASI (maintained silk elongation rate). By contrast, other regions influencing ASI had no influence on leaf growth. These results may have profound consequences for modelling the genotype x environment interaction and for designing drought-tolerant ideotypes. PMID- 17130184 TI - Cofilin plays a critical role in IL-8-dependent chemotaxis of neutrophilic HL-60 cells through changes in phosphorylation. AB - Cofilin is a ubiquitous, actin-binding protein. Only unphosphorylated cofilin binds actin and severs or depolymerizes filamentous actin (F-actin), and the inactive form of cofilin is phosphorylated at Ser 3. We reported recently that cofilin plays a regulatory role in superoxide production and phagocytosis by leukocytes, and in the present study, we investigated the role of cofilin in the chemotaxis of neutrophilic HL-60 cells. IL-8 is a potent, physiological chemokine, and it triggers a rapid, transient increase in F-actin beneath the plasma membrane and rapid dephosphorylation and subsequent rephosphorylation of cofilin. In this study, cofilin phosphorylation was found to be inhibited by S3-R peptide, which consists of a peptide corresponding to part of the phosphorylation site of cofilin and a membrane-permeable arginine polymer. When S3-R peptide was introduced into the neutrophilic cells, their chemotactic activity was enhanced, whereas a control peptide that contained an inverted sequence of the phosphorylation site of cofilin had no enhancing effect. Cofilin small interfering RNA (siRNA) decreased cofilin expression by about half and inhibited chemotaxis. In IL-8-stimulated cells, unphosphorylated cofilin accumulated around F-actin, and colocalization of F-actin and phosphorylated cofilin was observed, but these changes in cofilin localization were less prominent in cofilin siRNA treated cells. The inhibitors of PI-3K wortmannin and LY294002 inhibited the chemotaxis and suppressed IL-8-evoked dephosphorylation and rephosphorylation of cofilin. These results suggested that unphosphorylated cofilin plays a critical role in leukocyte chemotaxis and that PI-3K is involved in the control of the phosphorylation/dephosphorylation cycle of cofilin. PMID- 17130186 TI - Endocrine-related resources from the national institutes of health. PMID- 17130187 TI - Accuracy and predictive value of classification schemes for ketosis-prone diabetes. AB - OBJECTIVE: Ketosis-prone diabetes (KPD) is an emerging, heterogeneous syndrome. A sound classification scheme for KPD is essential to guide clinical practice and pathophysiologic studies. Four schemes have been used and are based on immunologic criteria, immunologic criteria and insulin requirement, BMI, and immunologic criteria and beta-cell function (Abeta classification). The aim of the present study is to compare the four schemes for accuracy and predictive value in determining whether KPD patients have absent or preserved beta-cell function, which is a strong determinant of long-term insulin dependence and clinical phenotype. RESEARCH DESIGN AND METHODS: Consecutive patients (n = 294) presenting with diabetic ketoacidosis and followed for 12-60 months were classified according to all four schemes. They were evaluated longitudinally for beta-cell autoimmunity, clinical and biochemical features, beta-cell function, and insulin dependence. beta-Cell function was defined by peak plasma C-peptide response to glucagon >or=1.5 ng/ml. The accuracy of each scheme to predict absent or preserved beta-cell function after 12 months of follow-up was tested using multiple statistical analyses. RESULTS: The "Abeta" classification scheme was the most accurate overall, with a sensitivity and specificity of 99.4 and 95.9%, respectively, positive and negative likelihood ratios of 24.55 and 0.01, respectively, and an area under the receiver operator characteristic curve of 0.972. CONCLUSIONS: The Abeta scheme has the highest accuracy and predictive value in classifying KPD patients with regard to clinical outcomes and pathophysiologic subtypes. PMID- 17130188 TI - Action or inaction? Decision making in patients with diabetes and elevated blood pressure in primary care. AB - OBJECTIVE: Hypertension increases micro- and macrovascular complications of diabetes. The goal for blood pressure is <130/80 mmHg. In primary care, however, blood pressure in many patients exceeds this goal. In this study, we evaluated the clinical decision-making process when a patient with diabetes presents with elevated blood pressure. RESEARCH DESIGN AND METHODS: Twenty-six primary care practices in two practice-based research networks in Colorado participated. Questionnaires were completed after each encounter with an adult with type 2 diabetes. Data obtained from the survey included 1) demographic information, 2) blood pressure results, 3) action taken, 4) type of action if action was taken, and 5) reasons for inaction if action was not taken. Bivariate and multivariate analyses were performed to identify predictors of action. RESULTS: Completed surveys totaled 778. Blood pressure was 130/74 +/- 18.8/12.0 mmHg (mean +/- SD). Sixty-two percent of patients exceeded goals. Action was taken to lower blood pressure in 34.9% of those. Predictors of action were 1) blood pressure level, 2) total number of medicines the patient was taking, and 3) patient already taking medicines for blood pressure. As blood pressure rose, providers attributed inaction more often to "competing demands" and reasons other than "blood pressure being at or near goal." CONCLUSIONS: No evidence was found for patterns of poor care among primary care physicians. Providers balance the clinical circumstances, including how elevated the blood pressure is, and issues of polypharmacy, medication side effects, and costs when determining the best course of action. Knowledge deficit is not a common cause of inaction. PMID- 17130189 TI - Community center-based resistance training for the maintenance of glycemic control in adults with type 2 diabetes. AB - OBJECTIVE: The purpose of this study was to determine whether beneficial effects on glycemic control of an initial laboratory-supervised resistance training program could be sustained through a community center-based maintenance program. RESEARCH DESIGN AND METHODS: We studied 57 overweight (BMI >or=27 kg/m2) sedentary men and women aged 40-80 years with established (>6 months) type 2 diabetes. Initially, all participants attended a twice-weekly 2-month supervised resistance training program conducted in the exercise laboratory. Thereafter, participants undertook a resistance training maintenance program (2 times/week) for 12 months and were randomly assigned to carry this out either in a community fitness and recreation center (center) or in their domestic environment (home). Glycemic control (HbA1c [A1C]) was assessed at 0, 2, and 14 months. RESULTS: Pooling data from the two groups for the 2-month supervised resistance training program showed that compared with baseline, mean A1C fell by -0.4% [95% CI -0.6 to -0.2]. Within-group comparisons showed that A1C remained lower than baseline values at 14 months in the center group (-0.4% [-0.7 to -0.03]) but not in the home group (-0.1% [-0.4 to 0.3]). However, no between-group differences were observed at each time point. Changes in A1C during the maintenance period were positively associated with exercise adherence in the center group only. CONCLUSIONS: Center-based but not home-based resistance training was associated with the maintenance of modestly improved glycemic control from baseline, which was proportional to program adherence. Our findings emphasize the need to develop and test behavioral methods to promote healthy lifestyles including increased physical activity in adults with type 2 diabetes. PMID- 17130190 TI - Differences in glucose tolerance between fixed-dose antihypertensive drug combinations in people with metabolic syndrome. AB - OBJECTIVE: We sought to test the hypothesis that a fixed-dose combination of trandolapril/verapamil-SR (T/V) is superior to a fixed-dose combination of losartan/hydrochlorothiazide (L/H) on glucose tolerance in hypertensive patients with impaired glucose tolerance (IGT). RESEARCH DESIGN AND METHODS: A prospective, randomized, open-label, blinded-end points design was used to assess the effects of a T/V versus L/H combination in patients with IGT and hypertension (n = 240) followed for up to 1 year. Doses were titrated to a systolic blood pressure <130 mmHg. Primary outcome was change from baseline in a 2-h glucose on oral glucose tolerance test (OGTT) at study end (mean [+/-SD] at follow-up, 46.9 +/- 13.5 weeks). Secondary outcomes included changes in insulin sensitivity, office and 24-h ambulatory blood pressure, incidence of new-onset diabetes, lipids, and inflammatory markers. Data are expressed as means +/- SE unless otherwise noted. RESULTS: Changes at study end were noted in 2-h OGTT glucose (T/V -0.21 +/- 0.36 vs. L/H +1.44 +/- 0.36 mmol/l; P < 0.001) and insulin level ( 30.13 +/- 38.38 vs. +84.86 +/- 38.33 pmol/l, respectively; P = 0.025). Worsening of insulin resistance occurred by week 12 (T/V 0.000 +/- 0.001 vs. L/H -0.005 +/- 0.001; P = 0.016). A higher incidence of new-onset diabetes (T/V 11.0 vs. L/H 26.6%; P = 0.002) and HbA1c >7% (2.6 vs. 9.6%, respectively; P = 0.05) occurred at study end. CONCLUSIONS: In patients with IGT, normal kidney function, and hypertension, the fixed-dose combination of T/V reduces the risk of new-onset diabetes compared with an L/H-based therapy. PMID- 17130191 TI - Beneficial associations of physical activity with 2-h but not fasting blood glucose in Australian adults: the AusDiab study. AB - OBJECTIVE: We examined the associations of physical activity with fasting plasma glucose (FPG) and with 2-h postload plasma glucose (2-h PG) in men and women with low, moderate, and high waist circumference. RESEARCH DESIGN AND METHODS: The Australian Diabetes, Obesity and Lifestyle (AusDiab) study provided data on a population-based cross-sectional sample of 4,108 men and 5,106 women aged >or=25 years without known diabetes or health conditions that could affect physical activity. FPG and 2-h PG were obtained from an oral glucose tolerance test. Self reported physical activity level was defined according to the current public health guidelines as active (>or=150 min/week across five or more sessions) or inactive (<150 min/week and/or less than five sessions). Sex-specific quintiles of physical activity time were used to ascertain dose response. RESULTS: Being physically active and total physical activity time were independently and negatively associated with 2-h PG. When physical activity level was considered within each waist circumference category, 2-h PG was significantly lower in active high-waist circumference women (beta -0.30 [95% CI -0.59 to -0.01], P = 0.044) and active low-waist circumference men (beta -0.25 [-0.49 to -0.02], P = 0.036) compared with their inactive counterparts. Considered across physical activity and waist circumference categories, 2-h PG levels were not significantly different between active moderate-waist circumference participants and active low waist circumference participants. Associations between physical activity and FPG were nonsignificant. CONCLUSIONS: There are important differences between 2-h PG and FPG related to physical activity. It appears that 2-h PG is more sensitive to the beneficial effects of physical activity, and these benefits occur across the waist circumference spectrum. PMID- 17130192 TI - Age, sex, and ethnic variations in serum insulin concentrations among U.S. youth: findings from the National Health and Nutrition Examination Survey 1999-2002. AB - OBJECTIVE: Distributions of serum concentrations of insulin among adolescents and young adults are poorly understood in the U.S. The objective of this study was to describe the distribution of serum insulin across demographic characteristics of U.S. adolescents and young adults. RESEARCH DESIGN AND METHODS: A total of 1,791 male and female subjects aged 12-19 years who participated in the National Health and Nutrition Examination Surveys for 1999-2002 were included in the analyses. RESULTS: Among male participants, serum concentrations of insulin increased from age 12 to 14 years before decreasing. Among female participants, concentrations were highest at age 13 years before decreasing steadily through age 19 years. Among participants aged 12-17 years but not those aged 18-19 years, females had higher mean log-transformed concentrations than males (P, Wald, F = 0.038 and 0.125, respectively) after adjusting for age and ethnicity. After adjusting for age and BMI percentile, mean log-transformed concentrations were higher in African-American females aged 12-17 years than in white or Mexican-American participants. No significant ethnic differences were found among female participants aged 18-19 years or male participants aged 12-19 years. Concentrations of insulin increased strongly with increasing levels of BMI. CONCLUSIONS: These results provide detailed information about serum concentrations of insulin in a representative sample of U.S. adolescents and young adults and may be useful to monitor future trends of this risk factor for diabetes and cardiovascular disease. PMID- 17130193 TI - HbA1c in early diabetic pregnancy and pregnancy outcomes: a Danish population based cohort study of 573 pregnancies in women with type 1 diabetes. AB - OBJECTIVE: To assess the association between first-trimester HbA(1c) (A1C) and the risk of adverse pregnancy outcomes in type 1 diabetic pregnancies. RESEARCH DESIGN AND METHODS: We identified all pregnant diabetic women in a Danish county from 1985 to 2003. A1C values from first trimester were collected, and pregnancy outcome was dichotomized as good (i.e., babies surviving the 1st month of life without major congenital abnormalities) and adverse (i.e., spontaneous and therapeutic abortion, stillbirth, neonatal death, or major congenital abnormalities detected within the 1st month). The prevalence of adverse outcomes was calculated according to quintiles of A1C. We computed receiver operating characteristic and lowess curve estimates and fitted logistic regression models to calculate prevalence odds ratio while adjusting for confounding by White class and smoking status. RESULTS: Of 573 pregnancies, 165 (29%) terminated with adverse outcomes. The prevalence of adverse outcomes varied sixfold from 12% (95% CI 7.2-17) in the lowest to 79% (60-91) in the highest quintile of A1C exposure. From A1C levels >7%, we found an almost linear association between A1C and risk of adverse outcome, whereby a 1% increase in A1C corresponded to 5.5% (3.8-7.3) increased risk of adverse outcome. CONCLUSIONS: Starting from a first-trimester A1C level slightly <7%, there is a dose-dependent association between A1C and the risk of adverse pregnancy outcome without indication of a plateau, below which the association no longer exits. A1C, however, seems to be of limited value in predicting outcome in the individual pregnancy. PMID- 17130194 TI - Patient Interpretation of Neuropathy (PIN) questionnaire: an instrument for assessment of cognitive and emotional factors associated with foot self-care. AB - OBJECTIVE: Using the common-sense model of illness behavior, we developed and validated a self-report instrument for assessment of patients' cognitive and emotional representations of diabetic peripheral neuropathy (DPN) influencing foot self-care. RESEARCH DESIGN AND METHODS: The Patient Interpretation of Neuropathy (PIN) questionnaire, generated from discussions with clinicians and interviews with patients with DPN, was administered to patients with DPN attending U.K. (n = 325) and U.S. (n = 170) diabetes centers. Psychometric tests of the PIN questionnaire comprised factor analysis, internal consistency, and test-retest reliability. Partial correlations and multivariate regressions established construct and criterion-related validity. The associations of PIN scales to past foot ulceration and foot self-care behaviors were compared with those using a generic measure of illness perception and emotion, the Revised Illness Perception Questionnaire (IPQ-R), which was adapted to neuropathy. RESULTS: Factor analysis of the PIN questionnaire produced 11 scales, which explained 69% of item variance. Nine factors measured patients' common-sense beliefs about DPN and their levels of understanding of DPN-related medical information. Two factors assessed the emotions of worry about potential consequences and anger at practitioners. Most scales demonstrated adequate internal (Cronbach's alpha = 0.62-0.90) and test-retest reliability (Pearson's r = 0.51-0.64). Partial correlations between the PIN and IPQ-R scales in corresponding domains were significant but modest (rp = 0.15-0.26). Finally, PIN scales showed significant associations with past foot ulceration and foot self care behaviors, thereby confirming criterion validity. CONCLUSIONS: The 39-item PIN questionnaire is a reliable and valid measure of patients' cognitive and emotional representations of neuropathy affecting foot self-care. PMID- 17130195 TI - Long-term effect of the Internet-based glucose monitoring system on HbA1c reduction and glucose stability: a 30-month follow-up study for diabetes management with a ubiquitous medical care system. AB - OBJECTIVE: To investigate the long-term effectiveness of the Internet-based glucose monitoring system (IBGMS) on glucose control in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: We conducted a prospective, randomized, controlled trial in 80 patients with type 2 diabetes for 30 months. The intervention group was treated with the IBGMS, while the control group made conventional office visits only. HbA1c (A1C) was performed at 3-month intervals. For measuring of the stability of glucose control, the SD value of A1C levels for each subject was used as the A1C fluctuation index (HFI). RESULTS: The mean A1C and HFI were significantly lower in the intervention group (n = 40) than in the control group (n = 40). (A1C [mean +/- SD] 6.9 +/- 0.9 vs. 7.5 +/- 1.0%, P = 0.009; HFI 0.47 +/- 0.23 vs. 0.78 +/- 0.51, P = 0.001; intervention versus control groups, respectively). Patients in the intervention group with a basal A1C >or=7% (n = 27) had markedly lower A1C levels than corresponding patients in the control group during the first 3 months and maintained more stable levels throughout the study (P = 0.022). Control patients with a basal A1C <7% (n = 15) showed the characteristic bimodal distribution of A1C levels, whereas the A1C levels in the intervention group remained stable throughout the study with low HFI. CONCLUSIONS: Long-term use of the IBGMS has proven to be superior to conventional diabetes care systems based on office visits for controlling blood glucose and achieving glucose stability. PMID- 17130196 TI - Effect of the dipeptidyl peptidase-4 inhibitor sitagliptin as monotherapy on glycemic control in patients with type 2 diabetes. AB - OBJECTIVE: To examine the efficacy and safety of once-daily oral sitagliptin as monotherapy in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: In a randomized, double-blind, placebo-controlled study, 741 patients (baseline HbA(1c) [A1C] 8.0%) were randomized to sitagliptin 100 or 200 mg or placebo for 24 weeks. RESULTS: Sitagliptin 100 and 200 mg produced significant (P < 0.001) placebo-subtracted reductions in A1C (-0.79 and -0.94%, respectively) and fasting plasma glucose (-1.0 mmol/l [-17.1 mg/dl] and -1.2 mmol/l [-21.3 mg/dl], respectively). Patients with baseline A1C >or=9% had greater reductions in placebo-subtracted A1C with sitagliptin 100 and 200 mg (-1.52 and -1.50%, respectively) than those with baseline A1C <8% (-0.57 and -0.65%) or >or=8 to <9.0% (-0.80 and -1.13%, respectively). In a meal tolerance test, sitagliptin 100 and 200 mg significantly decreased 2-h postprandial glucose (PPG) (placebo subtracted PPG -2.6 mmol/l [-46.7 mg/dl] and -3.0 mmol/l [-54.1 mg/dl], respectively). Results for the above key efficacy parameters were not significantly different between sitagliptin doses. Homeostasis model assessment of beta-cell function and proinsulin-to-insulin ratio improved with sitagliptin. The incidence of hypoglycemia was similar, and overall gastrointestinal adverse experiences were slightly higher with sitagliptin. No meaningful body weight changes from baseline were observed with sitagliptin 100 (-0.2 kg) or 200 mg ( 0.1 kg). The body weight change with placebo (-1.1 kg) was significantly (P < 0.01) different from that observed with sitagliptin. CONCLUSIONS: In this 24-week study, once-daily sitagliptin monotherapy improved glycemic control in the fasting and postprandial states, improved measures of beta-cell function, and was well tolerated in patients with type 2 diabetes. PMID- 17130197 TI - Efficacy and safety of the dipeptidyl peptidase-4 inhibitor sitagliptin added to ongoing metformin therapy in patients with type 2 diabetes inadequately controlled with metformin alone. AB - OBJECTIVE: The efficacy and safety of the dipeptidyl peptidase-4 inhibitor, sitagliptin, added to ongoing metformin therapy, were assessed in patients with type 2 diabetes who had inadequate glycemic control (HbA(1c) [A1C] >or=7 and or=1,500 mg/day) were randomly assigned to receive the addition of placebo or sitagliptin 100 mg once-daily in a 1:2 ratio for 24 weeks. Patients exceeding specific glycemic limits were provided rescue therapy (pioglitazone) until the end of the study. The efficacy analyses were based on an all-patients-treated population using an ANCOVA and excluded data obtained after glycemic rescue. RESULTS: At week 24, sitagliptin treatment led to significant reductions compared with placebo in A1C (-0.65%), fasting plasma glucose, and 2-h postmeal glucose. Fasting insulin, fasting C-peptide, fasting proinsulin-to-insulin ratio, postmeal insulin and C-peptide areas under the curve (AUCs), postmeal insulin AUC-to glucose AUC ratio, homeostasis model assessment of beta-cell function, and quantitative insulin sensitivity check index were significantly improved with sitagliptin relative to placebo. A significantly greater proportion of patients achieved an A1C <7% with sitagliptin (47.0%) than with placebo (18.3%). There was no increased risk of hypoglycemia or gastrointestinal adverse experiences with sitagliptin compared with placebo. Body weight decreased similarly with sitagliptin and placebo. CONCLUSIONS: Sitagliptin 100 mg once-daily added to ongoing metformin therapy was efficacious and well tolerated in patients with type 2 diabetes who had inadequate glycemic control with metformin alone. PMID- 17130198 TI - Relationship of fasting and hourly blood glucose levels to HbA1c values: safety, accuracy, and improvements in glucose profiles obtained using a 7-day continuous glucose sensor. AB - OBJECTIVE: In this study, we evaluated the safety and efficacy of 7-day transcutaneous, real-time, continuous glucose monitoring (CGM) in subjects with insulin-requiring diabetes. RESEARCH DESIGN AND METHODS: Eighty-six subjects were enrolled at five U.S. centers. Subjects wore a sensor inserted under the skin of the abdomen for 7 days during each of three consecutive periods. Data were blinded during period 1 and unblinded during periods 2 and 3. RESULTS: Of the 6,811 matched self-monitoring of blood glucose to sensor values prospectively analyzed, 97.2% fell in the Clarke error grid zones A and B, and median absolute relative difference was 11.4%. After unblinding, subjects reduced time spent at <55 mg/dl by 0.3 h/day, reduced time spent at >240 mg/dl by 1.5 h/day, and increased time in the target zone (81-140 mg/dl) by 1.4 h/day (P < 0.05 for all three comparisons). Improvements were seen in both types 1 and 2 diabetes and with use of both multiple daily injections and continuous subcutaneous insulin infusion. Modal day graphs were generated in six groups of subjects based on HbA1c (A1C) (10%). Mean glucose levels from midnight to 7:00 a.m. (fasting and dawn phenomenon periods) were only normal for subjects with A1C or=1 microg/g creatinine (3.34 [1.17-9.53]); the OR of patients currently smoking was 3.51 (1.14-10.80) relative to that of those who had never smoked. CONCLUSIONS: This study proves that the presence of MT-Ab can potentiate tubular dysfunction among diabetic subjects and that patients with high MT-Ab levels are more prone to development of tubular damage. PMID- 17130206 TI - Hyperinsulinemia and cognitive decline in a middle-aged cohort. AB - OBJECTIVE: Determining modifiable risks factors for cognitive decline and dementia are a public health priority as we seek to prevent dementia. Type 2 diabetes and related disorders such as hyperinsulinemia increase with aging and are increasing in the U.S. population. Our objective was to determine whether hyperinsulinemia is associated with cognitive decline among middle-aged adults without type 2 diabetes, dementia, or stroke in the Atherosclerosis Risk in Communities (ARIC) cohort. RESEARCH DESIGN AND METHODS: Middle-aged adults (aged 45-64 years at baseline) in the ARIC cohort had fasting insulin and glucose assessed between 1987 and 1989. Subjects with dementia, type 2 diabetes, or stroke at baseline were excluded from analysis. Three tests of cognitive function available at baseline and 6 years later were delayed word recall (DWR), digit symbol subtest (DSS), and first letter word fluency (WF). Cross-sectional comparisons and linear regression models were computed for cognitive tests at baseline and change in cognitive test scores to determine whether cognitive function was associated with two measures of insulin resistance, fasting insulin and homeostasis model assessment (HOMA). Linear regression models controlled for age, sex, race, marital status, education level, smoking status, alcohol use, depression, hypertension, and hyperlipidemia. RESULTS: In unadjusted and adjusted analyses, hyperinsulinemia based on fasting insulin and HOMA at baseline was associated with significantly lower baseline DWR, DSS, and WF scores and a greater decline over 6 years in DWR and WF. CONCLUSIONS: Insulin resistance is a potentially modifiable midlife risk factor for cognitive decline and dementia. PMID- 17130207 TI - Household wealth and the metabolic syndrome in the Whitehall II study. AB - OBJECTIVE: The metabolic syndrome is more common in socially disadvantaged groups. Inequalities in household wealth are currently widening and may contribute to the increasing prevalence of the metabolic syndrome. RESEARCH DESIGN AND METHODS: This was a cross-sectional analysis of 1,509 women and 4,090 men (aged 45.2-68.9 years) of an occupational cohort study of 20 civil service departments located in London, U.K. Components of the metabolic syndrome were measured in 1997-1999 and defined using a modified World Health Organization definition. RESULTS: Own income, household income, and wealth were each strongly and inversely associated with the metabolic syndrome in both sexes (P(trend) < 0.001). Within each group of household wealth, the prevalence of the metabolic syndrome was higher in men than in women. Sex differences became smaller with decreasing household wealth, with the prevalence of the metabolic syndrome rising from 12.0 and 5.7% in the wealthiest men and women, respectively, to corresponding values of 23.6 and 20.1% in the poorest group. The odds ratio (95% CI) associated with each decrease of one category in household wealth was 1.25 (1.03-1.50) in men and 1.69 (1.18-2.41) in women, adjusting for age, household members, occupational grade, education, father's social class, personal and household income, ethnic group, smoking, alcohol intake, diet, and physical activity. CONCLUSIONS: Household wealth, a measure of assets accumulated over decades and generations, is strongly and inversely associated with the metabolic syndrome. Future research should explore the potential mechanisms by which wealth inequalities are associated with the metabolic syndrome. PMID- 17130208 TI - The metabolic syndrome is a risk indicator of microvascular and macrovascular complications in diabetes: results from Metascreen, a multicenter diabetes clinic based survey. AB - OBJECTIVE: We aimed at assessing the degree of association and the predictive power of the metabolic syndrome with regard to clinically detectable complications in patients with diabetes. RESEARCH DESIGN AND METHODS: Metascreen is a cross-sectional survey of metabolic syndrome and clinically detected diabetes complications performed in 8,497 patients (7,859 with type 2 diabetes and 638 with type 1 diabetes) randomly chosen in 176 diabetes outpatient clinics throughout Italy. The metabolic syndrome was defined according to either the American Heart Association/National Heart, Lung, and Blood Institute (AHA/NHLBI) or the International Diabetes Federation (IDF) diagnostic criteria. Multivariate analyses of the association(s) between either AHA/NHLBI or IDF metabolic syndrome and clinical complications were performed. Receiver-operator characteristic (ROC) curves were constructed to compare the predictive power of the two sets of diagnostic criteria of the metabolic syndrome. RESULTS: Either definition of the metabolic syndrome was an independent statistical indicator of the presence of nephropathy and neuropathy (P < 0.02-0.01) in type 1 diabetes and of all complications (P < 0.0001), including cardiovascular disease and retinopathy, in type 2 diabetes. For each complication, the ROC curves based on either AHA/NHLBI or IDF metabolic syndrome were similar to each other and to the ROC curves constructed with all continuous traits compounding the metabolic syndrome. CONCLUSIONS: The metabolic syndrome, defined according to AHA/NHLBI or IDF diagnostic criteria, is an independent clinical indicator and may be involved in the pathogenesis of both macro- and microvascular complications of diabetes. PMID- 17130209 TI - Prediction of postprandial glycemic exposure: utility of fasting and 2-h glucose measurements alone and in combination with assessment of body composition, fitness, and strength. AB - OBJECTIVE: To determine the best predictors of total postprandial glycemic exposure and peak glucose concentrations in nondiabetic humans. RESEARCH DESIGN AND METHODS: Data from 203 nondiabetic volunteers who ingested a carbohydrate containing mixed meal were analyzed. RESULTS: Fasting glucose and insulin concentrations were poor predictors of postprandial glucose area above basal (R2 = approximately 0.07, P < 0.001). The correlation was stronger for 2-h glucose concentration (R2 = 0.55, P < 0.001) and improved slightly but significantly (P < 0.001) with the addition of fasting glucose, insulin, age, sex, and body weight to the model (r2 = 0.58). The 2-h glucose concentration also predicted the peak glucose concentration (R2 = 0.37, P < 0.001) with strength of the prediction increasing (P < 0.001) modestly with the addition of fasting glucose, insulin, age, sex, and body weight to the model (R2 = 0.48, P < 0.001). On the other hand, addition of measures of body function and composition did not improve prediction of total glycemic exposure or peak glucose concentration. CONCLUSIONS: Isolated measures of fasting or 2-h glucose concentrations alone or in combination with more complex measures of body composition and function are poor predictors of postprandial glycemic exposure or peak glucose concentration. This may explain, at least in part, the weak and at times inconsistent relationship between these parameters and cardiovascular risk. PMID- 17130210 TI - Changes in inflammatory cytokines are related to impaired glucose tolerance in offspring of type 2 diabetic subjects. AB - OBJECTIVE: We sought to determine whether levels of inflammatory markers and different cytokines are abnormal in nondiabetic offspring of type 2 diabetic subjects. RESEARCH DESIGN AND METHODS: Cytokine levels were measured in 19 healthy control subjects and 129 offspring of patients with type 2 diabetes (109 with normal glucose tolerance [NGT] and 20 with impaired glucose tolerance [IGT]). Insulin sensitivity was determined with the hyperinsulinemic-euglycemic clamp, insulin secretion with the intravenous glucose tolerance test, and abdominal fat distribution with computed tomography. RESULTS: Levels of C reactive protein and inflammatory cytokines were elevated in nondiabetic offspring of type 2 diabetic subjects. Interleukin (IL)-1beta was increased in the NGT group and decreased in the IGT group. In contrast, levels of IL-1 receptor antagonist (IL-1Ra) were increased in both groups. IL-1beta and -Ra levels correlated inversely (P < 0.05) with rates of whole-body glucose uptake and IL-1beta positively with visceral fat mass (P < 0.05) in normoglycemic offspring. CONCLUSIONS: Nondiabetic offspring of type 2 diabetic subjects have changes in the levels of inflammatory cytokines. The level of IL-1Ra seems to be the most sensitive marker of cytokine response in the pre-diabetic state. PMID- 17130211 TI - Protein hydrolysate/leucine co-ingestion reduces the prevalence of hyperglycemia in type 2 diabetic patients. PMID- 17130212 TI - Identification of obesity in patients with type 2 diabetes from Australian primary care: the NEFRON-5 study. PMID- 17130213 TI - Underreporting of food intake in obese diabetic and nondiabetic patients. PMID- 17130214 TI - Risk of diabetes associated with prescribed glucocorticoids in a large population. PMID- 17130215 TI - Improved glycemic control in poorly controlled patients with type 1 diabetes using real-time continuous glucose monitoring. PMID- 17130216 TI - New insights on the simultaneous assessment of insulin sensitivity and beta-cell function with the HOMA2 method. PMID- 17130217 TI - Sympathetic overactivity, endothelial dysfunction, inflammation, and metabolic abnormalities cluster in grade III (World Health Organization) obesity: reversal through sustained weight loss obtained with laparoscopic adjustable gastric banding. PMID- 17130218 TI - Hyperglycemic crises in adult patients with diabetes: a consensus statement from the American Diabetes Association. PMID- 17130219 TI - Weight control in individuals with diabetes. PMID- 17130220 TI - Ketosis-prone type 2 diabetes: time to revise the classification of diabetes. PMID- 17130221 TI - Multifactorial interventions before laser photocoagulation improve outcome of diabetic macular edema. PMID- 17130222 TI - Type III allergy to insulin detemir: response to Darmon et al. PMID- 17130223 TI - Diabetes and driving. PMID- 17130224 TI - Continuous glucose monitoring system with an alarm: a tool to reduce hypoglycemic episodes in pregnancy with diabetes. PMID- 17130225 TI - Waist-to-height ratio and BMI predict different cardiovascular risk factors in Chinese children. PMID- 17130226 TI - Prevalence and associations of binge eating disorder in a multiethnic population with type 2 diabetes. PMID- 17130227 TI - Translating the chronic care model into the community: results from a randomized controlled trial of a multifaceted diabetes care intervention: response to Piatt et al. PMID- 17130228 TI - Intensive insulin therapy in the intensive care unit: assessment by continuous glucose monitoring: response to De Block et al. PMID- 17130232 TI - Chromium picolinate supplementation attenuates body weight gain and increases insulin sensitivity in subjects with type 2 diabetes: response to Martin et al. PMID- 17130233 TI - Second harmonic generation confocal microscopy of collagen type I from rat tendon cryosections. AB - We performed second harmonic generation (SHG) imaging of collagen in rat-tendon cryosections, using femtosecond laser scanning confocal microscopy, both in backscattering and transmission geometries. SHG transmission images of collagen fibers were spatially resolved due to a coherent, directional SHG component. This effect was enhanced with the use of an index-matching fluid (n(i) = 1.52). The average SHG intensity oscillated with wavelength in the backscattered geometry (isotropic SHG component), whereas the spectral profile was consistent with quasi phase-matching conditions in transmission geometry (forward propagating, coherent SHG component) around 440 nm (lambda(p) = 880 nm). Collagen type I from bovine Achilles tendon was imaged for SHG in the backscattered geometry and its first order effective nonlinear coefficient was determined (|d(eff)| approximately 0.085(+/-0.025)x10(-12)mV(-1)) by comparison to samples of inorganic materials with known effective nonlinear coefficients (LiNbO3 and LiIO3). The SHG spectral response of collagen type I from bovine Achilles tendon matched that of the rat tendon cryosections in backscattered geometry. Collagen types I, II, and VI powders (nonfibrous) did not show any detectable SHG, indicating a lack of noncentrosymmetric crystalline structure at the molecular level. The various stages of collagen thermal denaturation were investigated in rat-tendon cryosections using SHG and bright-field imaging. Thermal denaturation resulted in the gradual destruction of the SHG signal. PMID- 17130234 TI - Caspase-3 regulates catalytic activity and scaffolding functions of the protein tyrosine phosphatase PEST, a novel modulator of the apoptotic response. AB - The protein tyrosine phosphatase PEST (PTP-PEST) is involved in the regulation of the actin cytoskeleton. Despite the emerging functions attributed to both PTPs and the actin cytoskeleton in apoptosis, the involvement of PTP-PEST in apoptotic cell death remains to be established. Using several cell-based assays, we showed that PTP-PEST participates in the regulation of apoptosis. As apoptosis progressed, a pool of PTP-PEST localized to the edge of retracting lamellipodia. Expression of PTP-PEST also sensitized cells to receptor-mediated apoptosis. Concertedly, specific degradation of PTP-PEST was observed during apoptosis. Pharmacological inhibitors, immunodepletion experiments, and in vitro cleavage assays identified caspase-3 as the primary regulator of PTP-PEST processing during apoptosis. Caspase-3 specifically cleaved PTP-PEST at the (549)DSPD motif and generated fragments, some of which displayed increased catalytic activity. Moreover, caspase-3 regulated PTP-PEST interactions with paxillin, leupaxin, Shc, and PSTPIP. PTP-PEST acted as a scaffolding molecule connecting PSTPIP to additional partners: paxillin, Shc, Csk, and activation of caspase-3 correlated with the modulation of the PTP-PEST adaptor function. In addition, cleavage of PTP-PEST facilitated cellular detachment during apoptosis. Together, our data demonstrate that PTP-PEST actively contributes to the cellular apoptotic response and reveal the importance of caspases as regulators of PTPs in apoptosis. PMID- 17130235 TI - Suppression of beta-amyloid precursor protein signaling into the nucleus by estrogens mediated through complex formation between the estrogen receptor and Fe65. AB - The C-terminal fragment of the beta-amyloid precursor protein produced after cleavage by gamma-secretase, namely, APPct or AICD, has been shown to form a multimeric complex with the adaptor protein Fe65 and to regulate transcription through the recruitment of the histone acetyltransferase Tip60. The present study shows that 17beta-estradiol inhibits the transcriptional and apoptotic activities of the APPct complex by a process involving the interaction of estrogen receptor alpha (ERalpha) with Fe65. ERalpha-Fe65 complexes were detected both in vitro and in the mouse brain, and recruitment of ERalpha to the promoter of an APPct target gene (KAI1) was demonstrated. Our studies reveal a novel mechanism of estrogen action, which may explain the well-known neuroprotective functions of estrogens as well as the complex role of this female hormone in the pathogenesis of neuronal degeneration diseases. PMID- 17130236 TI - Targeted disruption of the murine retinal dehydrogenase gene Rdh12 does not limit visual cycle function. AB - RDH12 codes for a member of the family of short-chain alcohol dehydrogenases/reductases proposed to function in the visual cycle that supplies the chromophore 11-cis retinal to photoreceptor cells. Mutations in RDH12 cause severe and progressive childhood onset autosomal-recessive retinal dystrophy, including Leber congenital amaurosis. We generated Rdh12 knockout mice, which exhibited grossly normal retinal histology at 10 months of age. Levels of all trans and 11-cis retinoids in dark- and light-adapted animals and scotopic and photopic electroretinogram (ERG) responses were similar to those for the wild type, as was recovery of the ERG response following bleaching, for animals matched for an Rpe65 polymorphism (p.L450M). Lipid peroxidation products and other measures of oxidative stress did not appear to be elevated in Rdh12(-/-) animals. RDH12 was localized to photoreceptor inner segments and the outer nuclear layer in both mouse and human retinas by immunohistochemistry. The present findings, together with those of earlier studies showing only minor functional deficits in mice deficient for Rdh5, Rdh8, or Rdh11, suggest that the activity of any one isoform is not rate limiting in the visual response. PMID- 17130237 TI - Nucleolin is required for RNA polymerase I transcription in vivo. AB - Eukaryotic genomes are packaged with histones and accessory proteins in the form of chromatin. RNA polymerases and their accessory proteins are sufficient for transcription of naked DNA, but not of chromatin, templates in vitro. In this study, we purified and identified nucleolin as a protein that allows RNA polymerase II to transcribe nucleosomal templates in vitro. As immunofluorescence confirmed that nucleolin localizes primarily to nucleoli with RNA polymerase I, we demonstrated that nucleolin allows RNA polymerase I transcription of chromatin templates in vitro. The results of chromatin immunoprecipitation experiments established that nucleolin is associated with chromatin containing rRNA genes transcribed by RNA polymerase I but not with genes transcribed by RNA polymerase II or III. Knockdown of nucleolin by RNA interference resulted in specific inhibition of RNA polymerase I transcription. We therefore propose that an important function of nucleolin is to permit RNA polymerase I to transcribe nucleolar chromatin. PMID- 17130238 TI - Role of phosphocholine cytidylyltransferase alpha in lung development. AB - Lung development depends upon the differentiation and expansion of a variety of specialized epithelial cell types, including distal type I and type II pneumocytes in the late term. Previous studies have shown a strict dependence on the choline cytidylyltransferase alpha isoform (CCTalpha) to mediate membrane phospholipid formation in cultured cells and during preimplantation embryogenesis. CCTalpha expression is highest in lung, and there has long been speculation about its precise role, due to the dual requirement for phospholipid in proliferating cell membranes and for lung surfactant production from alveolar type II cells. We investigated the function of CCTalpha in lung development, using an inducible, epithelial cell-specific CCTalpha knockout mouse line. Deletion of CCTalpha beginning at embryonic day 7.5 did not restrict lung development but resulted in severe respiratory failure at birth. Alveolar lavage and lung lipid analyses showed significant decreases in the major surfactant phospholipid, dipalmitoyl-phosphatidylcholine. The fatty acids destined for the surfactant phospholipid were redirected to an expanded triglyceride pool. Transcripts encoding type II cell-specific markers were expressed in the knockout mice, indicating the expected progression of differentiation in lung epithelia. However, surfactant protein levels were reduced, with the exception of that for surfactant protein B, which was elevated. Ultrastructural analysis of the type II cells showed Golgi complex abnormalities and aberrant lamellar bodies, which deliver surfactant lipid and protein to the alveolar lumen. Thus, CCTalpha was not required for the proliferation or differentiation of lung epithelia but was essential for the secretory component of phospholipid synthesis and critical for the proper formation of lamellar bodies and surfactant protein homeostasis. PMID- 17130239 TI - Inactivation of CUG-BP1/CELF1 causes growth, viability, and spermatogenesis defects in mice. AB - CUG-BP1/CELF1 is a multifunctional RNA-binding protein involved in the regulation of alternative splicing and translation. To elucidate its role in mammalian development, we produced mice in which the Cugbp1 gene was inactivated by homologous recombination. These Cugbp1(-/-) mice were viable, although a significant portion of them did not survive after the first few days of life. They displayed growth retardation, and most Cugbp1(-/-) males and females exhibited impaired fertility. Male infertility was more thoroughly investigated. Histological examination of testes from Cugbp1(-/-) males showed an arrest of spermatogenesis that occurred at step 7 of spermiogenesis, before spermatid elongation begins, and an increased apoptosis. A quantitative reverse transcriptase PCR analysis showed a decrease of all the germ cell markers tested but not of Sertoli and Leydig markers, suggesting a general decrease in germ cell number. In wild-type testes, CUG-BP1 is expressed in germ cells from spermatogonia to round spermatids and also in Sertoli and Leydig cells. These findings demonstrate that CUG-BP1 is required for completion of spermatogenesis. PMID- 17130240 TI - The human SETMAR protein preserves most of the activities of the ancestral Hsmar1 transposase. AB - Transposons have contributed protein coding sequences to a unexpectedly large number of human genes. Except for the V(D)J recombinase and telomerase, all remain of unknown function. Here we investigate the activity of the human SETMAR protein, a highly expressed fusion between a histone H3 methylase and a mariner family transposase. Although SETMAR has demonstrated methylase activity and a DNA repair phenotype, its mode of action and the role of the transposase domain remain obscure. As a starting point to address this problem, we have dissected the activity of the transposase domain in the context of the full-length protein and the isolated transposase domain. Complete transposition of an engineered Hsmar1 transposon by the transposase domain was detected, although the extent of the reaction was limited by a severe defect for cleavage at the 3' ends of the element. Despite this problem, SETMAR retains robust activity for the other stages of the Hsmar1 transposition reaction, namely, site-specific DNA binding to the transposon ends, assembly of a paired-ends complex, cleavage of the 5' end of the element in Mn(2+), and integration at a TA dinucleotide target site. SETMAR is unlikely to catalyze transposition in the human genome, although the nicking activity may have a role in the DNA repair phenotype. The key activity for the mariner domain is therefore the robust DNA-binding and looping activity which has a high potential for targeting the histone methylase domain to the many thousands of specific binding sites in the human genome provided by copies of the Hsmar1 transposon. PMID- 17130241 TI - Mitotic Cdc6 stabilizes anaphase-promoting complex substrates by a partially Cdc28-independent mechanism, and this stabilization is suppressed by deletion of Cdc55. AB - Ectopic expression of Cdc6p results in mitotic delay, and this has been attributed to Cdc6p-mediated inhibition of Cdc28 protein kinase and failure to activate the anaphase-promoting complex (APC). Here we show that endogenous Cdc6p delays a specific subset of mitotic events and that Cdc28 inhibition is not sufficient to account for it. The depletion of Cdc6p in G(2)/M cells reveals that Cdc6p is rate limiting for the degradation of the APC/Cdc20 substrates Pds1p and Clb2p. Conversely, the premature expression of Cdc6p delays the degradation of APC/Cdc20 substrates. Abolishing Cdc6p/Cdc28p interaction does not eliminate the Cdc6-dependent delay of these anaphase events. To identify additional Cdc6 mediated, APC-inhibitory mechanisms, we looked for mutants that reversed the mitotic delay. The deletion of SWE1, RAD24, MAD2, or BUB2 had no effect. However, disrupting CDC55, a PP2A regulatory subunit, suppressed the Cdc6p-dependent delay of Pds1 and Clb2 destruction. A specific role for CDC55 was supported by demonstrating that the lethality of Cdc6 ectopic expression in a cdc16-264 mutant is suppressed by the deletion of CDC55, that endogenous Cdc6p coimmunoprecipitates with the Cdc55 and Tpd3 subunits of PP2A, that Cdc6p/Cdc55p/Tpd3 interaction occurs only during mitosis, and that Cdc6 affects PP2A-Cdc55 activity during anaphase. This demonstrates that the levels and timing of accumulation of Cdc6p in mitosis are appropriate for mediating the modulation of APC/Cdc20. PMID- 17130242 TI - Molecular analysis of fibulin-5 function during de novo synthesis of elastic fibers. AB - Elastic fibers contribute to the structural support of tissues and to the regulation of cellular behavior. Mice deficient for the fibulin-5 gene (fbln5(-/ )) were used to further elucidate the molecular mechanism of elastic fiber assembly. Major elastic fiber components were present in the skin of fbln5(-/-) mice despite a dramatic reduction of mature elastic fibers. We found that fibulin 5 preferentially bound the monomeric form of elastin through N-terminal and C terminal elastin-binding regions and to a preexisting matrix scaffold through calcium-binding epidermal growth factor (EGF)-like (CB-EGF) domains. We further showed that adenovirus-mediated gene transfer of fbln5 was sufficient to regenerate elastic fibers and increase elastic fiber-cell connections in vivo. A mutant fibulin-5 lacking the first 28 amino acids of the first CB-EGF domain, however, was unable to rescue elastic fiber defects. Fibulin-5 thus serves as an adaptor molecule between monomeric elastin and the matrix scaffold to aid in elastic fiber assembly. These results also support the potential use of fibulin-5 as a therapeutic agent for the treatment of elastinopathies. PMID- 17130243 TI - Murine CXCL14 is dispensable for dendritic cell function and localization within peripheral tissues. AB - Dendritic cells (DCs) have long been recognized as key regulators of immune responses. However, the process of their recruitment to peripheral tissues and turnover during homeostasis remains largely unknown. The chemokine CXCL14 (BRAK) is constitutively expressed in skin and other epithelial tissues. Recently, the human chemokine was proposed to play a role in the homeostatic recruitment of macrophage and/or DC precursors toward the periphery, such as skin. Although so far no physiological function could be demonstrated for the murine CXCL14, it shows a remarkable homology to the human chemokine. In order to elucidate the in vivo role of CXCL14, we generated a mouse defective for this chemokine. We studied various components of the immune system with emphasis on monocytes/macrophages and DC/Langerhans cell (LC) populations in different tissues during steady state but did not find a significant difference between knockout (CXCL14(-)(/)(-)) and control mice. Functionally, LCs were able to become activated, to migrate out of skin, and to elicit a delayed type of hypersensitivity reaction. Overall, our data indicate that murine CXCL14 is dispensable for the homeostatic recruitment of antigen-presenting cells toward the periphery and for LC functionality. PMID- 17130244 TI - Murine Pif1 interacts with telomerase and is dispensable for telomere function in vivo. AB - Pif1 is a 5'-to-3' DNA helicase critical to DNA replication and telomere length maintenance in the budding yeast Saccharomyces cerevisiae. ScPif1 is a negative regulator of telomeric repeat synthesis by telomerase, and recombinant ScPif1 promotes the dissociation of the telomerase RNA template from telomeric DNA in vitro. In order to dissect the role of mPif1 in mammals, we cloned and disrupted the mPif1 gene. In wild-type animals, mPif1 expression was detected only in embryonic and hematopoietic lineages. mPif1(-/-) mice were viable at expected frequencies, displayed no visible abnormalities, and showed no reproducible alteration in telomere length in two different null backgrounds, even after several generations. Spectral karyotyping of mPif1(-/-) fibroblasts and splenocytes revealed no significant change in chromosomal rearrangements. Furthermore, induction of apoptosis or DNA damage revealed no differences in cell viability compared to what was found for wild-type fibroblasts and splenocytes. Despite a novel association of mPif1 with telomerase, mPif1 did not affect the elongation activity of telomerase in vitro. Thus, in contrast to what occurs with ScPif1, murine telomere homeostasis or genetic stability does not depend on mPif1, perhaps due to fundamental differences in the regulation of telomerase and/or telomere length between mice and yeast or due to genetic redundancy with other DNA helicases. PMID- 17130245 TI - Recurrent compartment syndrome after fracture of a tibiofibular synostosis in a National Football League player. PMID- 17130246 TI - Arthroscopic distal clavicle resection in athletes: a prospective comparison of the direct and indirect approach. AB - BACKGROUND: The clinical success of arthroscopic distal clavicle resection for athletes has been well documented. There are, however, no published studies that prospectively compare the recovery rates in athletes as well as the outcomes of the indirect versus direct approaches. HYPOTHESIS: Both procedures are equally successful; however, the direct approach affords faster return to sports. STUDY DESIGN: Randomized controlled clinical trial; Level of evidence, 2. METHODS: Thirty-eight consecutive athletes with osteolysis of the distal clavicle or isolated posttraumatic arthrosis of the acromioclavicular joint without instability underwent arthroscopic distal clavicle resection. The patients were randomized into 2 groups: a direct superior approach and an indirect subacromial approach. American Shoulder and Elbow Surgeons and Athletic Shoulder Scoring System scores were measurable outcomes. RESULTS: Thirty-four athletes were available for a minimum 2-year follow-up. The 2 groups were similar, including preoperative American Shoulder and Elbow Surgeons and Athletic Shoulder Scoring System scores. Both groups demonstrated significant improvement in both scores at final follow-up when compared with preoperative scores (P < .001). The direct group demonstrated higher American Shoulder and Elbow Surgeons (82 vs 64) and Athletic Shoulder Scoring System (74 vs 56) scores at week 2 (P < .001) and week 6 (American Shoulder and Elbow Surgeons, 88 vs 77; Athletic Shoulder Scoring System, 87 vs 73) (P < .001). At final follow-up, both groups demonstrated excellent clinical outcomes, even though there was a statistical difference in scores, with the direct group scoring better (American Shoulder and Elbow Surgeons, 95.7 vs 91.2; Athletic Shoulder Scoring System -94.9 vs 88.3). The direct group demonstrated faster return to sports (mean, 21 days) than the indirect group (mean, 42 days) (P < .001). Radiographic analysis demonstrated an equivalent resection. One patient in each group had a clinically insignificant increase in coracoclavicular distance. CONCLUSIONS: Both the direct superior approach and the indirect subacromial approach to the arthroscopic distal clavicle resection result in successful clinical outcome with clinically insignificant difference at final follow-up. Athletes treated with the direct superior approach improved faster clinically and returned to sports earlier. PMID- 17130247 TI - Laxity testing of the shoulder: a review. AB - Laxity testing is an important part of the examination of any joint. In the shoulder, it presents unique challenges because of the complexity of the interactions of the glenohumeral and scapulothoracic joints. Many practitioners believe that laxity testing of the shoulder is difficult, and they are unclear about its role in evaluation of patients. The objectives of the various laxity and instability tests differ, but the clinical signs of such tests can provide helpful information about joint stability. This article summarizes the principles of shoulder laxity testing, reviews techniques for measuring shoulder laxity, and evaluates the clinical usefulness of the shoulder laxity tests. Shoulder laxity evaluation can be a valuable element of the shoulder examination in patients with shoulder pain and instability. PMID- 17130248 TI - Effects of racial discrimination and health behaviors on mental and physical health of middle-class African American men. AB - This research is an examination of the effects of racial discrimination and health-promoting behaviors on the physical and mental health of a sample of 399 well-educated African American men. One would think that the attainment of higher education would increase health-promoting behaviors and might decrease discriminatory experiences that impact health. However, regression analysis indicated a more complex picture. Health-promoting behaviors were positively related to mental health, whereas experiences of racial discrimination contributed to poorer mental health. Relationships between health-promoting behaviors and that of racial discrimination to physical health were found to be nonsignificant. In conclusion, the authors discuss the importance of culturally appropriate health-promotion efforts. PMID- 17130249 TI - Microplate orbital mixing improves high-throughput cell-based reporter assay readouts. AB - Reporter assays are commonly used for high-throughput cell-based screening of compounds, cDNAs, and siRNAs due to robust signal, ease of miniaturization, and simple detection and analysis. Among the most widely used reporter genes is the bioluminescent enzyme luciferase, which, when exposed to its substrate luciferin upon cell lysis, yields linear signal over a dynamic range of several orders of magnitude. Commercially available luciferase assay formulations have been developed permitting homogeneous, single-step cell lysis and reporter activity measurements. Assay conditions employed with these formulations are typically designed to minimize well-to-well luminescence variability due to variability in dispensing, evaporation, and incomplete sample mixing. The authors demonstrate that incorporating a microplate orbital mixing step into 96- and 384-well microplate cell-based luciferase reporter assays can greatly improve reporter readouts. They have found that orbital mixing using commercially available mixers facilitates maximal luciferase signal generation from high cell density containing samples while minimizing variability due to partial cell lysis, thereby improving assay precision. The authors fully expect that widespread availability of mixers with sufficiently small orbits and higher speed settings will permit gains in signal and precision in the 1536-well format as well. PMID- 17130250 TI - Optimizing classification of drug-drug interaction potential for CYP450 isoenzyme inhibition assays in early drug discovery. AB - In drug discovery, the potential of cytochrome P450 inhibition of new chemical entities is frequently quantified in terms of IC50 values. In early drug discovery, a risk classification into low, medium, or high potential inhibitors is often sufficient for ranking and prioritizing of compounds. Although often 6 or more inhibitor concentrations are used to determine the IC50 value, the question arises whether it is possible to predict the risk class based on fewer inhibitor concentrations with comparable reliability. In this article, the authors propose a new integrated 2-point method with inhibitor concentrations chosen in accordance with the risk classification. They analyze its predictive power and the feasibility of not only classifying the compounds into different risk classes but also ranking those compounds that have been binned into the middle risk class. The proposed integrated 2-point method is thus highly suitable for automation. Altogether, it maintains the quality of the prediction while considerably reducing time and cost. The proposed method is applicable to other IC50 assays and risk classifications. PMID- 17130251 TI - C57BL/6 and congenic interleukin-10-deficient mice can serve as models of Campylobacter jejuni colonization and enteritis. AB - Campylobacter jejuni is a globally distributed cause of human food-borne enteritis and has been linked to chronic joint and neurological diseases. We hypothesized that C. jejuni 11168 colonizes the gastrointestinal tract of both C57BL/6 mice and congenic C57BL/6 interleukin-10-deficient (IL-10(-/-)) mice and that C57BL/6 IL-10(-/-) mice experience C. jejuni 11168-mediated clinical signs and pathology. Individually housed mice were challenged orally with C. jejuni 11168, and the course of infection was monitored by clinical examination, bacterial culture, C. jejuni-specific PCR, gross pathology, histopathology, immunohistochemistry, and anti-C. jejuni-specific serology. Ceca of C. jejuni 11168-infected mice were colonized at high rates: ceca of 50/50 wild-type mice and 168/170 IL-10(-/-) mice were colonized. In a range from 2 to 35 days after infection with C. jejuni 11168, C57BL/6 IL-10(-/-) mice developed severe typhlocolitis best evaluated at the ileocecocolic junction. Rates of colonization and enteritis did not differ between male and female mice. A dose-response experiment showed that as little as 10(6) CFU produced significant disease and pathological lesions similar to responses seen in humans. Immunohistochemical staining demonstrated C. jejuni antigens within gastrointestinal tissues of infected mice. Significant anti-C. jejuni plasma immunoglobulin levels developed by day 28 after infection in both wild-type and IL-10-deficient animals; antibodies were predominantly T-helper-cell 1 (Th1)-associated subtypes. These results indicate that the colonization of the mouse gastrointestinal tract by C. jejuni 11168 is necessary but not sufficient for the development of enteritis and that C57BL/6 IL-10(-/-) mice can serve as models for the study of C. jejuni enteritis in humans. PMID- 17130252 TI - Gene expression pattern in human brain endothelial cells in response to Neisseria meningitidis. AB - To extend our knowledge of target proteins in endothelial cells infected with the meningitis-causing pathogen Neisseria meningitidis, we characterized the interaction between the bacterial and human brain microvascular endothelial cell (HBMEC) monolayers. By use of human cDNA microarrays, transcriptional analysis revealed distinct responses to 4 and 8 h of infection. We also addressed the question of whether the major virulence factor of meningococci, i.e., the capsule, influences the host cell response. Of the 1,493 (at 4 h postinfection) and 1,246 (at 8 h postinfection) genes with altered expression upon bacterial contact, about 49.4% and 45%, respectively, depended on capsule expression. In particular, we identified an increase of expression for genes encoding proteins involved in bacterial adhesion and invasion. High levels of apoptosis-related gene (bad, bak, asp, and immediate-early response gene 1) expression could also be detected in infected cells. Further analyses confirmed that HBMECs displayed several hallmarks of apoptosis in response to N. meningitidis infection, namely, phosphatidylserine translocation and activation of caspase 3 and AMP-activated protein kinase alpha. Moreover, several differentially regulated genes not previously known to respond to meningococcal infection were identified. Of these, genes encoding cell adhesion proteins (CD44, CD98, and CD99), genes involved in downstream signaling of integrins (integrin-linked kinase, mitogen-activated protein kinase kinase 1, and mitogen-activated protein kinase kinase kinase 10) as well as negative regulators of these pathways (dual-specificity phosphatases 1, 5, and 14 and G protein pathway suppressor 2), and genes involved in cytoskeleton reorganization (those encoding Arp2/3, p34-arc, actinin alpha 1, vasodilatator-stimulated protein, and Wiskott-Aldrich syndrome protein) were the most prominent. This global transcriptional analysis creates a new platform for further molecular and cellular analysis of the interaction between N. meningitidis and target cells. PMID- 17130254 TI - Hypertension and cardiovascular risk in chronic kidney disease patients. AB - This supplement of the Journal of American Society of Nephrology contains some of the proceedings of the Fifth International Conference on Hypertension and the Kidney. The Conference, held in Madrid, Spain, in February 2006, was organized by the Department of Nephrology of the Hospital General, Universitario Gregorio Maranon, under the sponsorship of the Universidad Complutense de Madrid, Spanish Society of Nephrology, Spanish Society of Hypertension, and European Renal Association-European Dialysis and Transplant Association. PMID- 17130253 TI - Phosphorylcholine decreases early inflammation and promotes the establishment of stable biofilm communities of nontypeable Haemophilus influenzae strain 86-028NP in a chinchilla model of otitis media. AB - Nontypeable Haemophilus influenzae (NTHi) is a leading causative agent of otitis media. Much of the inflammation occurring during NTHi disease is initiated by lipooligosaccharides (LOS) on the bacterial surface. Phosphorylcholine (PCho) is added to some LOS forms in a phase-variable manner, and these PCho(+) variants predominate in vivo. Thus, we asked whether this modification confers some advantage during infection. Virulence of an otitis media isolate (NTHi strain 86 028NP) was compared with that of an isogenic PCho transferase (licD) mutant using a chinchilla (Chinchilla lanigera) model of otitis media. Animals infected with NTHi 86-028NP licD demonstrated increased early inflammation and a delayed increase in bacterial counts compared to animals infected with NTHi 86-028NP. LOS purified from chinchilla-passed NTHi 86-028NP had increased PCho content compared to LOS purified from the inoculum. Both strains were recovered from middle ear fluids as long as 14 days postinfection. Biofilms were macroscopically visible in the middle ears of euthanized animals infected with NTHi 86-028NP 7 days and 14 days postchallenge. Conversely, less dense biofilms were observed in animals infected with NTHi 86-028NP licD 7 days postinfection, and none of the animals infected with NTHi 86-028NP licD had a visible biofilm by 14 days. Fluorescent antibody staining revealed PCho(+) variants within biofilms, similar to our prior results with tissue culture cells in vitro (S. L. West-Barnette, A. Rockel, and W. E. Swords, Infect. Immun. 74:1828-1836, 2006). Animals coinfected with equal proportions of both strains had equal persistence of each strain and somewhat greater severity of disease. We thus conclude that PCho promotes NTHi infection and persistence by reducing the host inflammatory response and by promoting formation of stable biofilm communities. PMID- 17130255 TI - Proteomic analysis of early left ventricular hypertrophy secondary to hypertension: modulation by antihypertensive therapies. AB - Untreated or poorly controlled arterial hypertension induced development of pathologic left ventricular hypertrophy (LVH), a common finding in hypertensive patients and a strong predictor of cardiovascular morbidity and mortality. The proteomic approach is a powerful technique to analyze a complex mixture of proteins in various settings. An experimental model of hypertension-induced early LVH was performed in spontaneously hypertensive rats, and the cardiac protein pattern compared with the normotensive Wistar Kyoto counterpart was analyzed. Fifteen altered protein spots were shown in the early stage of LVH. Compared with a previous animal model of established and regressed LVH, three protein spots were common in both models. These three altered protein spots corresponded to two unique proteins that were identified as Calsarcin-1 (CS-1) and ubiquinone biosynthesis protein COQ7 homolog. CS-1 is a negative regulator of the calcineurin/NF-AT pathway. Because upregulation in the expression levels of this protein was observed, the activation level of NF-kappaB by oxidative stress as an alternative pathway was investigated. It was found that antihypertensive therapies partially decreased oxidative stress and normalized the activation of NF-kappaB in the kidneys and aorta NF-kappaB activation but just moderately in the heart. This could be due to the interaction of any specific cardiac protein with any component of the NF-kappaB pathway. In this sense, CS-1 could be a good candidate because it is expressed preferentially in heart, to a lesser extent in smooth muscle cells, but not in kidney. Further investigations are necessary to elucidate the exact role of CS-1 and ubiquinone biosynthesis protein COQ7 in the setting of hypertension-induced LVH. PMID- 17130256 TI - Uric acid, the metabolic syndrome, and renal disease. AB - Metabolic syndrome, characterized by truncal obesity, hypertriglyceridemia, elevated BP, and insulin resistance, is recognized increasingly as a major risk factor for kidney disease and also is a common feature of patients who are on dialysis. One feature that is common to patients with metabolic syndrome is an elevated uric acid. Although often considered to be secondary to hyperinsulinemia, recent evidence supports a primary role for uric acid in mediating this syndrome. Specifically, fructose, which rapidly can cause metabolic syndrome in rats, also raises uric acid, and lowering uric acid in fructose-fed rats prevents features of the metabolic syndrome. Uric acid also can accelerate renal disease in experimental animals and epidemiologically is associated with progressive renal disease in humans. It is proposed that fructose and purine-rich foods that have in common the raising of uric acid may have a role in the epidemic of metabolic syndrome and renal disease that is occurring throughout the world. PMID- 17130257 TI - Dissecting inflammation in ESRD: do cytokines and C-reactive protein have a complementary prognostic value for mortality in dialysis patients? AB - Because atherogenesis represents a type of chronic inflammation that involves multiple elements of the inflammatory-immune response, the simultaneous prediction power for death of C-reactive protein (CRP) and proinflammatory cytokines (IL-1 beta, IL-6, IL-18, and TNF-alpha) was tested in a cohort of 217 patients with ESRD. During the follow-up period (average 41 mo), 112 patients died. In an analysis that was adjusted for other risk factors, the relative risks for death of patients who were exposed to high levels of one, two, three, and four or more inflammation biomarkers were 1.48, 1.64, 2.76, and 3.05 times higher, respectively, than that of patients in the reference category (no inflammation). In this model, the explained variation in mortality that was attributable to overall inflammation burden (+9.1%) was marginally higher (P = 0.06) than that provided by IL-6 alone (+6.1%). In an alternative analysis based on the Bayesian approach (receiver operating characteristic curves analysis), the prediction power of the combined inflammatory burden was identical to that provided by the sole IL-6 (0.59 +/- 0.04 versus 0.59 +/- 0.04). IL-6 captures almost entirely the prediction power of the overall inflammation burden in patients with ESRD. IL-6 seems to be an almost ideal indicator of the severity of inflammation. The use of this biomarker can be recommended in clinical studies that aim to better the understanding of inflammation or to modify it in this population. PMID- 17130258 TI - Oxidative stress in uremia: the role of anemia correction. AB - Patients with chronic kidney disease (CKD) are prone to develop cardiovascular disorders. Numerous reports have shown the association between uremia and oxidative stress, which increases patients' risk for cumulative injury to multiple organs. Anemia is a common and disabling feature of CKD and seems to be a main cause of oxidative stress; correction of anemia represents an effective approach to reduce oxidative stress and, consequently, cardiovascular risk. There is increasing evidence that correction of anemia with erythropoiesis-stimulating agents could protect from oxidative stress in patients with CKD and ESRD. However, iron deficiency frequently complicates anemia in patients with CKD, and ferrous iron cation is a co-factor that is needed for hydroxyl radical production, which can promote cytotoxicity and tissue injury. This has raised a justifiable concern that prescription of intravenous iron may exacerbate oxidative stress and, hence, endothelial dysfunction, inflammation, and progression of cardiovascular disease, which are widely known consequences of CKD. Correction of anemia represents an effective approach to reduce oxidative stress and, consequently, cardiovascular risk. Iron deficiency is a common cause of resistance to erythropoiesis-stimulating agents, and the overall risk-benefit ratio favors use of intravenous iron to treat iron deficiency in patients with CKD. Consecutive or combined treatment with intravenous iron and erythropoiesis stimulating agents clearly is beneficial for patients with CKD and iron deficiency, and anemia and could contribute to prevent the risk for cardiovascular events in these patients. PMID- 17130259 TI - Prevalence of abnormal urinary albumin excretion rate in hypertensive patients with impaired fasting glucose and its association with cardiovascular disease. AB - The prevalence and significance of microalbuminuria in hypertensive patients with impaired fasting glucose (IFG) has received very little attention. A total of 10,320 hypertensive patients who attended primary care centers were enrolled in this study, and the final analysis was done in 7625 patients: 1459 without IFG (plasma glucose <100 mg/dl), 3010 with IFG (plasma glucose > or =100 mg/dl and <126 mg/dl), and 3156 with type 2 diabetes (plasma glucose >126 mg/dl). Microalbuminuria was determined using the Micro Albustix reactive strip from Bayer (high urinary albumin excretion [UAE]: Albumin/creatinine ratio > or =3.4 mg/mmol). The proportion of patients with high UAE was 39.4, 48.3, and 65.6%, respectively, in the three groups (P < 0.01 for the trend). The differences in UAE between the group with IFG and the group with normal fasting glucose persisted after adjustment for age, gender, systolic BP, fasting plasma glucose, and cardiovascular comorbidity (odds ratio 1.74; 95% confidence interval 1.08 to 2.80). Hypertensive patients with IFG and high UAE showed a higher prevalence of ischemic heart disease, cardiac insufficiency, left ventricular hypertrophy, atrial fibrillation, and renal insufficiency than the group with normal UAE. Global prevalence of cardiovascular conditions was 30.4% in the group with high UAE compared with 21.4% in the group with normal UAE (odds ratio 1.60; 95% confidence interval 1.31 to 1.95). It is concluded that almost half of hypertensive patients with IFG have high UAE and a higher prevalence of associated cardiovascular involvement and renal insufficiency. PMID- 17130260 TI - Vascular protection of dual therapy (atorvastatin-amlodipine) in hypertensive patients. AB - Hypertension frequently coexists with other cardiovascular risk factors, such as hypercholesterolemia, and their combination is associated with a greater rate of cardiovascular events. Recent clinical data support that treatment of hypertensive patients with a combination of antihypertensive and lipid-lowering therapies leads to a higher reduction in the incidence of cardiovascular events. In the Anglo-Scandinavian Cardiac Outcomes Trial-Blood Pressure Lowering Arm (ASCOT-BPLA), an optimal prevention of cardiovascular events was reached in patients who were randomly assigned to atorvastatin and the amlodipine treatment. However, the potential underlying mechanisms of these vascular protective effects are not fully elucidated. Because experimental studies have shown that statins and calcium channel blockers have antiatherosclerotic effects, the effect of atorvastatin alone or in combination with amlodipine was analyzed in the protein secretion profile of atherosclerotic plaques that were cultured ex vivo. In this respect, the addition of atorvastatin and amlodipine to atherosclerotic plaques normalized the levels of the different released proteins to that obtained from healthy arteries. This review highlights recent clinical and experimental studies that support that a combined treatment of hypertensive patients with both statins and calcium channel blockers could promote a higher reduction in their global cardiovascular risk profile and associated mortality. As an example, the application of a proteomic approach to assess the modulation by atorvastatin alone or in combination with amlodipine on the proteins that are released by atherosclerotic plaques has allowed the identification of novel therapeutic targets by which these drugs could promote their additive/synergic effects. PMID- 17130261 TI - Prevalence of renal insufficiency in individuals with hypertension and obesity/overweight: the FATH study. AB - Overweight and obesity are associated with increased cardiovascular risk. Some studies have demonstrated that they also can result in renal damage. The aim of this study was to assess the prevalence of renal insufficiency (RI), defined as a GFR <60 ml/min per 1.73 m2, in a cohort of 4585 patients who attended primary care with essential hypertension and a body mass index > or =25 kg/m2. The patients were classified as overweight and obese according to body mass index (25 to 29.9 and > or =30 kg/m2, respectively). Abdominal obesity was defined as a waist circumference > or =88 and 102 cm in women and men, respectively. Both groups had a high prevalence of metabolic syndrome (Adult Treatment Panel III). The prevalence of RI was high in both the overweight group (22.7%; 95% confidence interval [CI] 20.6 to 24.9) and in the obese group (22.8%; 95% CI 21.0 to 24.7). The presence of diabetes increased the risk for RI (odds ratio 1.83; 95% CI 1.55 to 2.16). The prevalence of RI was greater in patients with abdominal obesity (23 versus 17%; P < 0.001). In the presence of abdominal obesity, cardiovascular risk factors and components of the metabolic syndrome also were more prevalent. The higher risk for RI with abdominal obesity persisted even after adjustment for dyslipidemia, elevated blood glucose levels, and other variables that are associated with RI (adjusted odds ratio 1.40; 95% CI 0.84 to 2.33). It was concluded that patients who have hypertension and visceral obesity and attend primary care present a higher prevalence of metabolic syndrome and RI. PMID- 17130262 TI - Relationship between ankle-brachial index and chronic kidney disease in hypertensive patients with no known cardiovascular disease. AB - Both decreased GFR and albuminuria are associated with an elevated prevalence of peripheral artery disease. However, the combined effects of these alterations previously were not evaluated. Patients with hypertension and with no known vascular disease (n = 955; mean age 66 yr; 56% male) were selected from internal medicine outpatient clinics throughout Spain. Cardiovascular risk factors, urinary albumin excretion, and the ankle-brachial index (ABI) were assessed in all participants. GFR was estimated according to the Cockroft-Gault equation. Of the study population, 62% had diabetes, 23.8% had a GFR <60 ml/min per 1.73 m2, and 43.8% had albuminuria. The prevalence of ABI <0.9 was greater in patients with a GFR <60 ml/min per 1.73 m2 (37.4 versus 24.3%; P < 0.0001) and in those who had albuminuria (32.2 versus 23.3%; P = 0.001). In patients with both alterations, the prevalence of ABI <0.9 was 45.7%. Multivariate analysis indicated that the factors that were associated independently with low ABI were age (odds ratio [OR] 1.06; 95% confidence interval [CI] 1.03 to 1.08; P < 0.0001), triglyceride concentration (OR 1.003; 95% CI 1.001 to 1.005; P = 0.001), presence of albuminuria (OR 1.61; 95% CI 1.18 to 2.20; P = 0.003), smoking habit (OR 1.72; 95% CI 1.13 to 2.63; P = 0.012), and a GFR <60 ml/min per 1.73 m2 (OR 1.47; 95% CI 1.01 to 2.17; P = 0.049). In patients with hypertension and without known vascular disease, reduced GFR and albuminuria are associated independently with an ABI <0.9. Their combined presence characterizes a subgroup of the population who have an elevated prevalence of peripheral artery disease and could benefit from early diagnosis and treatment. PMID- 17130263 TI - Insulin resistance, inflammatory biomarkers, and adipokines in patients with chronic kidney disease: effects of angiotensin II blockade. AB - Patients with chronic kidney disease (CKD) present a high prevalence of insulin resistance (IR). Some studies suggest that angiotensin II may influence some cellular pathways that contribute to the pathogenesis of IR and stimulate the release of proinflammatory cytokines. Fifty-two patients who had stages 3 and 4 CKD and no diabetes were administered an angiotensin receptor blocker (ARB), olmesartan (40 mg), for 16 wk. Before and after ARB treatment, metabolic and inflammatory parameters and adipokines were measured. IR was calculated by Homeostasis Model Assessment (HOMA) index. Baseline data were compared with data that were obtained from 25 healthy control individuals of similar age and normal renal function. Compared with control subjects, patients with CKD presented significantly higher BP and waist circumference, higher triglycerides and lower HDL levels, higher insulin levels, and higher mean HOMA index (6.0 +/- 2.7 versus 2.9 +/- 2.2 muU/ml x mmol/L; P < 0.001). In addition, patients with CKD had increased levels of high-sensitivity C-reactive protein, TNF-alpha, and IL-6. In patients with CKD, leptin was positively correlated to abdominal obesity, insulin levels, and IL-6, and adiponectin was inversely correlated to abdominal obesity and insulin levels. Olmesartan treatment resulted in a significant decrease of BP, urinary protein excretion, plasma glucose (99 +/- 16 versus 92 +/- 14 mg/dl; P < 0.05), insulin (23.1 +/- 8.8 versus 19.9 +/- 9; P < 0.05), HOMA index (6.0 +/ 2.7 versus 4.7 +/- 2.8; P < 0.05), and glycated hemoglobin (5.33 +/- 0.58 versus 4.85 +/- 0.81%; P < 0.01). At the same time, there was a significant reduction of high-sensitivity C-reactive protein levels, from 4.45 mg/L (2.45 to 9.00) to 3.55 mg/L (1.80 to 5.40; P < 0.05) and fibrinogen (412 +/- 100 versus 370 +/- 105 mg/dl; P < 0.05). There were no significant differences in adipokine levels after olmesartan treatment. These data demonstrate that patients with CKD have a high prevalence of IR, metabolic syndrome, and chronic inflammation and that the administration of the ARB olmesartan improves IR and inflammation markers in these patients. Plasma adipokine levels that are related to several metabolic risk factors in patients with CKD were not modified by ARB therapy. PMID- 17130264 TI - Effect of drastic weight loss after bariatric surgery on renal parameters in extremely obese patients: long-term follow-up. AB - Obesity is a health problem that is reaching epidemic proportions. Extreme obesity (body mass index [BMI] > or =40 kg/m2) is a type of obesity that usually does not respond to medical treatment, with surgery being the current treatment of choice. Extreme obesity is associated with cardiovascular disease, type 2 diabetes, dyslipidemia, and hypertension. Recently, obesity has been related with high rate of renal lesions, but renal function and renal parameters in extreme obesity scarcely are documented. The objective of this study was to evaluate the effect of weight loss after bariatric surgery (BS) on BP, renal parameters, and renal function in 61 extremely obese (EO) patients after 24 mo of follow-up. A total of 61 EO adults (37 women) were studied prospectively before and 24 mo after surgery. Control subjects were 24 healthy, normal-weight adults (15 women). Anthropometric, BP, and renal parameters were determined. Presurgery weight, BMI, GFR, 24-h proteinuria, and 24-h albuminuria were higher in the EO patients than in control subjects (P < 0.001). All parameters improved at 12 mo after BS. However, during the second year of follow-up, only 24-h albuminuria (P = 0.006) and BMI (P = 0.014) continued to improve. At 24 mo after BS, obesity-related renal alterations considerably improved. This improvement was observed mainly in the first year after surgery, when the majority of weight loss occurred. However, 24-h albuminuria still improves during the second year of follow-up. It is possible that this decrease in 24-h albuminuria is not GFR related but rather is attributable to the persistence of the decrease in BMI and to the improvement of other weight-related metabolic factors. PMID- 17130265 TI - Mycophenolate mofetil treatment improves hypertension in patients with psoriasis and rheumatoid arthritis. AB - Evidence that was obtained in several experimental models and in strains of hypertensive rats indicates that infiltration of inflammatory cells and oxidative stress in the kidney play a role in the induction and maintenance of hypertension. Similar evidence is lacking in human hypertension, at least in part, because immunosuppressive treatment is unjustified in patients with hypertension. For addressing this issue, patients who were prescribed by their private physicians mycophenolate mofetil (MMF) for the treatment of psoriasis or rheumatoid arthritis and had, in addition, grade I essential hypertension and normal renal function were studied. Eight patients were studied before MMF was started, during MMF treatment, and 1 mo after MMF treatment had been discontinued. Other treatments and diet were unchanged in the three phases of the study. MMF therapy was associated with a significant reduction in systolic, diastolic, and mean BP. Urinary excretion of TNF-alpha was reduced progressively by MMF treatment and increased after MMF was discontinued. Reduction of urinary malondialdehyde, TNF-alpha, and RANTES excretion during MMF administration did not reach statistical significance but had a direct positive correlation with the BP levels. These data are consistent with the hypothesis that renal immune cell infiltration and oxidative stress play a role in human hypertension. PMID- 17130266 TI - Lipid changes and statins in chronic renal insufficiency. AB - It has been known for a long time that chronic kidney disease (CKD) is associated with dyslipidemia, but the full extent of abnormalities has been appreciated only recently, because routine laboratory tests fail to disclose the entire spectrum of lipid abnormalities. Lipids, particularly HDL cholesterol, are predictive of cardiovascular events, but a paradoxic inverse relation between cholesterol concentration and cardiovascular death has been noted in uremic patients. This currently is thought to be explained by the confounding effect of microinflammation and possibly calcification, but this is not definitely proved. Several retrospective analyses that included patients with mild or moderate CKD documented benefit from lowering of cholesterol by statins. In contrast, the Die Deutsche Diabetes Dialyse (4D) study and a small Scandinavian study failed to show a benefit from lowering of cholesterol by statins in ESRD. Pathomechanistically, it is possible that nonclassical pathomechanisms override statin-sensitive mechanisms as also suggested by the observation that statins fail to reduce carotid intima-media thickening. Although, experimentally, exposure to lipids (particularly oxidized lipids) aggravates progression, data on the effect of statins on progression in patients with CKD are not definite. The most likely explanation is that the impact of numerous confounders obscures their effect on progression. The increase in urinary protein excretion of patients who are treated with statins had been a cause of concern, but the underlying mechanism (i.e. interference with proximal tubular reabsorption of protein) meanwhile has been well documented. PMID- 17130267 TI - Effects of atorvastatin on inflammatory and fibrinolytic parameters in patients with chronic kidney disease. AB - Although substantial evidence suggests that treatment of dyslipidemia with statins reduces mortality and morbidity that are associated with cardiovascular disease, only a few studies have examined the efficacy of statins on inflammatory and fibrinolytic status in patients with chronic kidney disease (CKD). A 6-mo, prospective, randomized study was designed to assess the efficacy of atorvastatin in reducing circulating inflammatory and fibrinolytic parameters in patients with CKD. Sixty-six patients with CKD (stages 2, 3, and 4) and LDL cholesterol levels > or =100 mg/dl were randomly assigned (2:1) to receive 20 mg/d atorvastatin (n = 44) or nonatorvastatin therapy (n = 22). Lipid profile, renal function, fibrinolytic balance (tissue plasminogen activator [t-PA] and plasminogen activator inhibitor-1), and inflammatory markers (C-reactive protein [CRP], IL-1 beta, IL-6, and TNF-alpha) were measured before and 6 mo after atorvastatin was added to the treatment. Twenty-five age-matched individuals with normal renal function (estimated GFR >90 ml/min) were used as healthy control subjects. Patients with CKD had higher CRP, IL-1 beta, TNF-alpha, and IL-6 levels than age matched population with normal renal function. t-PA concentration was higher in patients with CKD (P = 0.000). Plasminogen activator inhibitor-1 values were comparable in all patients. Total cholesterol and LDL cholesterol were significantly reduced only in patients who received atorvastatin. In addition to the hypolipidemic effect, atorvastatin treatment significantly reduced inflammatory parameters: CRP (median 4.1 to 2.9; P = 0.015), TNF-alpha (6.0 +/- 2.7 to 4.7 +/- 2.4; P = 0.046), and IL-1 beta levels (1.9 +/- 0.7 to 1.2 +/- 0.7; P = 0.001). These parameters remained unchanged in patients who were not treated with atorvastatin. Fibrinolytic parameters were not modified by atorvastatin treatment. Patients with CKD showed higher levels of inflammatory parameters and t-PA levels than age-matched healthy control subjects. Atorvastatin treatment, in addition to its beneficial effect on cholesterol levels, improved the inflammatory state of these patients without modifying fibrinolytic balance. PMID- 17130268 TI - Urinary albumin excretion and glomerular filtration rate across the spectrum of glucose abnormalities in essential hypertension. AB - The objective of this study was to assess the relationship between urinary albumin excretion (UAE) and GF across the spectrum of the glucose metabolism abnormalities in a large population of patients with hypertension. The Microaluminuria en Pacientes con Glucemia Basal Alterada (MAGAL) is a multicenter, cross-sectional study that was carried out by 1723 primary care physicians. A total of 6227 patients with essential hypertension (in three groups: [1] normal fasting glucose <100 mg/dl, [2] impaired fasting glucose > or =100 to 126 mg/dl, and [3] type 2 diabetes) were analyzed in this substudy. GFR was estimated by using the Modification of Diet in Renal Disease (MDRD) abbreviated equation. A single first-morning urine albumin/creatinine ratio was measured using Bayer reagent strip Microalbustix, a semiquantitative method. Abnormal UAE was defined as an albumin/creatinine ratio > or =3.4 mg/mmol (equivalent to > or =30 mg/g). The prevalence of abnormal UAE, > or =3.4 mg/mmol, increased across the spectrum of glucose abnormalities: 39.7, 46.2, 48.6, and 65.6% for normoglycemic, low-range, and high-range impaired fasting glucose and diabetes, respectively. UAE was positively related to SBP (P = 0.003) and inversely to GFR (P < 0.001). Renal insufficiency (GFR <60 ml/min per 1.73 m2) was present in 21.8% of the patients, more frequently older patients, women, and those with diabetes. The factors that were related to renal insufficiency were UAE > or =3.4 mg/mmol (odds ratio 1.86; 95% confidence interval 1.60 to 2.17) and diabetes (odds ratio 1.62; 95% confidence interval 1.29 to 2.04). There is a close relationship between abnormal UAE and renal insufficiency in essential hypertension. This is more marked in patients with diabetes and moderate in patients with high-range impaired fasting glucose. PMID- 17130269 TI - Role of pulse pressure on cardiovascular risk in chronic kidney disease patients. AB - Epidemiologic studies have emphasized the close relationship between high BP and cardiovascular disease (CVD). Recently published prospective studies have focus on systolic and pulse pressure (PP). Systolic BP seems to be a more important factor than diastolic BP on cardiovascular and all-cause mortality in older patients. PP reflects stiffness of the large arteries and increases with age. Increasingly, PP is recognized as an independent predictor of myocardial infarction, congestive heart failure, and cardiovascular death, even in hypertensive patients who undergo successful antihypertensive drug therapy, especially in older individuals. Chronic kidney disease (CKD) is a major public health problem. The progression of kidney disease and its associated cardiovascular complications are the major causes of morbidity and mortality. This holds true for all stages of kidney disease, including ESRD that requires renal replacement therapy. Most of the traditional CVD risk factors are highly prevalent in CKD, and several nontraditional factors also are associated with atherosclerosis in CKD. The burden of hypertension is present at all stages of CKD. Several studies have shown that PP is a reliable prognostic factor for mortality and CVD in patients who have CKD and are on hemodialysis and in renal transplant patients. The purpose of this review is to show the importance of PP on cardiovascular risk in patients with CKD, including kidney transplant recipients. PMID- 17130270 TI - Dual blockade of the renin-angiotensin system in the progression of renal disease: the need for more clinical trials. AB - There is clear evidence that pharmacologic blockade of the renin-angiotensin system (RAS) with angiotensin-converting enzyme inhibitors (ACEI) or angiotensin receptor blockers (ARB) reduces proteinuria and slows the progression of renal disease in diabetic and nondiabetic nephropathies, a beneficial effect that is not related to BP control. Some patients exhibit a significant beneficial response, whereas others do not. The absence of response may be explained by the incomplete blockade of the RAS obtained with ACEI. In the search of new alternatives that could improve the antiproteinuric and nephroprotective effects of RAS blockers, the association of ACEI and ARB might prove useful. ARB produces a complete blockade of the RAS. Several studies have shown a more marked antiproteinuric effect of the dual blockade of the RAS versus ACEI or ARB alone. A recent study also demonstrated that this more marked antiproteinuric effect is associated with less progression of renal disease in primary nondiabetic nephropathies despite a similar effect on BP. Until now, there has not been any reference to a beneficial effect on progression of the dual blockade in type 2 diabetic nephropathy, which is the most frequent cause of ESRD. A multicenter, prospective, open, active-controlled, and parallel-group trial was designed to compare the effects of an ACE inhibitor versus an ARB or its combination on renal disease progression, proteinuria, and cardiovascular events in type 2 diabetic nephropathy. PMID- 17130271 TI - Left ventricular hypertrophy: is hyperphosphatemia among dialysis patients a risk factor? AB - Cardiovascular disease occurs in ESRD patients at rates that are far higher than is seen in the general population, and cardiovascular deaths account for the majority of deaths among dialysis patients. Abnormal mineral metabolism is a novel cardiovascular risk factor among dialysis patients. Recently published results demonstrated that even with good control of BP and anemia, conventional hemodialysis is associated with significant left ventricular hypertrophy (LVH); however, daily hemodialysis was associated with a significant reduction in LV mass index (LVMI). Furthermore, it was shown that control of serum phosphorus correlates with the reduction in LVMI. These data suggest a novel mechanism for the deleterious effect of elevated serum phosphorus on cardiovascular outcomes among hemodialysis patients: LVH. Other investigators have noted an association of hyperphosphatemia and LVH; however, this study was the first to demonstrate that improvement in serum phosphorus is associated with reduction in LVM. In addition, it is shown that daily hemodialysis is an effective modality in improving serum phosphorus through significantly improved phosphorus removal. Elevated serum phosphorus leads to vascular calcification, which can lead to LVH by decreasing vascular compliance. However, our study showed an improvement in LVMI during a 12-mo period. Because vascular calcification is unlikely to remit over this time period, it is proposed that serum phosphorus has a reversible, cardiotoxic effect that leads to LVH that can be reversed successfully with good control of serum phosphorus. PMID- 17130272 TI - Anemia and cardiovascular risk: the lesson of the CREATE Trial. AB - Anemia has received increasing attention as an independent cardiovascular risk factor in patients with chronic kidney disease (CKD); a number of studies have highlighted its clear relationship with CKD mortality, because its impact on cardiac function leads to the development of left ventricular hypertrophy. However, despite the association between higher hemoglobin levels and better outcomes, a number of clinical studies have failed to demonstrate that fully correcting anemia has a positive effect on morbidity and mortality in patients with CKD. The Cardiovascular Reduction Early Anemia Treatment Epoetin beta (CREATE) study was designed from the hypothesis that, as anemia develops early in the course of CKD and nearly at the same time as cardiovascular disease, its earlier correction may provide better protection against the development of cardiovascular abnormalities. This randomized, multicenter, open-label, parallel group trial involved 603 patients who had moderate anemia (hemoglobin 11 to 12.5 g/dl) and stage 3 to 4 CKD (estimated GFR 15 to 35 ml/min) and were randomly assigned to attain complete or partial anemia correction. The final results are due to be published within a few months, but the preliminary analyses do not show that complete anemia correction leads to any cardiovascular advantage, although the cardiovascular event rate was half that expected, possibly as a result of patient selection, trial effect, and improved medical care. The baseline findings also indicated that the burden of cardiovascular disease already is very high even in relatively early stages of CKD. PMID- 17130273 TI - Vascular calcifications: pathogenesis, management, and impact on clinical outcomes. AB - The predisposition to vascular calcifications in patients with chronic kidney disease (CKD) has gained great interest in recent years as many studies have described its likely impact on morbidity and mortality. The mechanism by which the process of vascular calcification is produced is complex, and it does not consist in a simple precipitation of calcium and phosphate but is instead an active and modifiable process. Several "modifiable and nonmodifiable" factors that are able to promote vascular calcification are extremely frequent in patients with CKD. Most of the present strategies to decrease vascular calcifications are based in the control of the more prevalent modifiable risk factors. Unfortunately, the extremely important nonmodifiable risk factors, which are highly prevalent, such as older age, time on dialysis, and diabetes, are not under one's control. Recent studies also have shown that vascular calcifications in some localizations were associated with increased osteoporotic fractures not only in dialysis patients but also in the general population, and interestingly, mortality also was associated significantly and positively with vascular calcifications and nontraumatic bone fractures. Despite that new strategies may improve the management of vascular diseases and specifically have a positive impact on the high prevalence of vascular calcifications, still the best possible control of the bone metabolic and inflammatory parameters are in the primary line. The horizon of the coming decade looks promising, but solid clinical and epidemiologic data are needed to manage better the bone- and cardiovascular related disorders in patients with CKD. PMID- 17130274 TI - Inflammatory syndrome in patients on hemodialysis. AB - Mortality is markedly elevated in hemodialysis (HD) patients. Between 30 and 50% of prevalent patients have elevated serum levels of inflammatory markers such as C-reactive protein and IL-6. The presence of inflammation, chronic or episodic, has been found to be associated with increased mortality risk. The causes of inflammation are multifactorial and include patient-related factors, such as underlying disease, comorbidity, oxidative stress, infections, obesity, and genetic or immunologic factors, or on the other side, HD-related factors, mainly depending on the membrane biocompatibility and dialysate quality. The adequate knowledge of these causes and their prevention or treatment if possible may contribute to improving the inflammatory state of patients who are on HD and possibly their mortality. PMID- 17130275 TI - Impact of treatment with calcimimetics on hyperparathyroidism and vascular mineralization. AB - Soft tissue calcification that involves primarily the medial portion of the arterial vasculature is a widely recognized and common complication of chronic kidney disease Vascular calcification (VC) causes increased arterial stiffness and contributes to the high cardiovascular mortality and morbidity in dialysis patients. The pathogenesis of VC is complex and includes factors that promote calcification and others that inhibit calcification. Studies in dialysis patients have shown a correlation between VC and a number of uremia-related factors. Overall, abnormalities in calcium and phosphate metabolism, such as hyperphosphatemia and a raised serum calcium-phosphorus product traditionally have been thought of as important determinants in patients with chronic renal failure. Common therapeutic interventions in secondary hyperparathyroidism have come under scrutiny for associations with the development of VC. Calcimimetics provide a means of controlling serum levels of parathyroid hormone in secondary hyperparathyroidism without increasing the calcium-phosphorus product and, more important, may lower the risk for VC in these patients. PMID- 17130276 TI - Ischemic heart disease after renal transplantation in patients on cyclosporine in Spain. AB - Ischemic heart disease (IHD), more common among transplant recipients than in the general population, accounts for approximately 50% of cardiovascular deaths. Despite its importance, only a few publications have addressed the prevalence of and risk factors for this complication. This was a retrospective cohort study in 2382 cadaver renal transplant recipients who were treated with cyclosporine as initial immunosuppression. Two groups were formed. The first group consisted of 163 patients with IHD, and the second group consisted of 326 patients without IHD. The prevalence of IHD was 6.8%, and the incidence was 15.7/1000 patient years. Cardiac events presented during the first year in 62 (38%) patients. Multivariate analysis showed that the risk factors for IHD were age at transplant in years (relative risk [RR] 1.054; 95% confidence interval [CI] 1.033 to 1.075; P = 0.000), male gender (RR 1.940; 95% CI 1.221 to 3.081; P = 0.005), body weight at transplant in kg (RR 1.020; 95% CI 1.007 to 1.033; P = 0.002), pretransplantation cardiovascular disease (RR 2.150; 95% CI 1.733 to 3.359; P = 0.001), and a history of pretransplantation hypercholesterolemia (RR 2.032; 95% CI 1.378 to 2.998; P = 0.000). When only ischemic events that occurred 12 mo after transplantation were taken into consideration, the risk factors were age, male gender, body weight, smoking, and pretransplantation and posttransplantation hypercholesterolemia, whereas pretransplantation cardiovascular disease disappeared from the model. IHD affected nearly 7% of transplant recipients. Smoking, hypertension, and hypercholesterolemia constituted the treatable risk factors for IHD in this population. Emphasis should be placed on the need to stop smoking and to control hypertension and pre- and posttransplantation levels of serum cholesterol. PMID- 17130277 TI - New-onset diabetes after kidney transplantation: risk factors. AB - New-onset diabetes after transplantation (NODAT) contributes to the risk for cardiovascular disease and infection, reducing graft and patient survival. For improvement of the outcome of kidney transplant recipients, it is of great interest to know precisely the risk factors that contribute to NODAT development. Nonmodifiable risk factors for development of NODAT are age, race, genetic background, family history of diabetes, and previous glucose intolerance. Modifiable risk factors are obesity and overweight, hepatitis C virus and cytomegalovirus infections, and immunosuppressive drugs. Both steroids and calcineurin inhibitors influence the appearance of NODAT, whereas the role of sirolimus in glucose metabolism currently is controversial. PMID- 17130278 TI - Impact of tacrolimus and mycophenolate mofetil combination on cardiovascular risk profile after kidney transplantation. AB - Cardiovascular risk factors after kidney transplantation are enhanced as a result of the chronic use of immunosuppressants. Tacrolimus with mycophenolate mofetil has become the most commonly used combination after kidney transplantation. Cardiovascular risk factors that are related to the use of this combined therapy have been analyzed in various clinical trials in comparison with other immunosuppressive therapies. This review summarizes the main results of these studies regarding arterial hypertension, lipid profile, posttransplantation diabetes, renal function, and even acute rejection rate. The aim is to characterize the cardiovascular risk profile of tacrolimus and mycophenolate mofetil association when compared with older and newer immunosuppressive associations. PMID- 17130279 TI - The role of CYP2A6 in the emergence of nicotine dependence in adolescents. AB - OBJECTIVES: The objectives of our study were to evaluate whether genetic variation in nicotine metabolic inactivation accounted for the emergence of nicotine dependence from mid- to late adolescence and whether initial smoking experiences mediated this effect. METHODS: Participants were 222 adolescents of European ancestry who participated in a longitudinal cohort study of the biobehavioral determinants of adolescent smoking. Survey data were collected annually from grade 9 to the end of grade 12. Self-report measures included nicotine dependence, smoking, age first smoked, initial smoking experiences, peer and household member smoking, and alcohol and marijuana use. DNA collected via buccal swabs was assessed for CYP2A6 alleles that are common in white people and are demonstrated to decrease enzymatic function (CYP2A6*2, *4, *9, *12). RESULTS: Latent growth-curve modeling indicated that normal metabolizers (individuals with no detected CYP2A6 variants) progressed in nicotine dependence at a faster rate and that these increases in nicotine dependence leveled off more slowly compared with slower metabolizers (individuals with CYP2A6 variants). Initial smoking experiences did not account for how CYP2A6 genetic variation impacts nicotine dependence. CONCLUSIONS: These findings may help to promote a better understanding of the biology of smoking behavior and the emergence of nicotine dependence in adolescents and inform future work aimed at understanding the complex interplay between genetic, social, and psychological factors in adolescent smoking behavior. PMID- 17130280 TI - Clinical disease in children associated with newly described coronavirus subtypes. AB - OBJECTIVES: Coronaviruses cause upper respiratory illness and occasionally lower tract disease in susceptible populations. In this study we examined the prevalence of 4 human coronaviruses, including subtypes OC43, 229E, and the recently described NL63 and HKU1 in a pediatric population presenting to a children's hospital. PATIENTS AND METHODS: Specimens collected over a 1-year period from pediatric patients presenting with acute respiratory illness were analyzed for the presence of 4 coronavirus subtypes using consensus and subtype specific real-time reverse-transcription polymerase chain reaction assays. The demographic and clinical characteristics associated with coronavirus infection were examined retrospectively. RESULTS: Coronaviruses were detected in 66 of 1043 children. Eight, 11, 19, and 28 specimens were positive for subtypes 229E, NL63, OC43, and HKU1, respectively. Coronaviruses were detected throughout the study period; all 4 of the subtypes were present simultaneously in December. The acute clinical features were similar across subtypes. Of 32 children infected with a coronavirus as the sole respiratory pathogen, 13 had lower respiratory tract disease. Children whose only detectable respiratory virus was a coronavirus were more likely to have underlying chronic disease than were children coinfected with another respiratory virus. CONCLUSIONS: Although 4 subtypes of coronavirus were detected, the recently discovered coronavirus subtypes NL63 and HKU1 accounted for the majority of coronaviruses detected in our cohort of mostly hospitalized children with respiratory symptoms. New subtypes likely represent a substantial portion of previously unexplained respiratory illnesses. PMID- 17130281 TI - The effects of oxandrolone and exercise on muscle mass and function in children with severe burns. AB - OBJECTIVES: Severe burns are associated with a significant loss of muscle and strength. Studies have reported that oxandrolone improves lean body mass in muscle-wasting conditions. Also shown previously in burned children is that an exercise program increases lean body mass and muscle strength. We hypothesized that oxandrolone, in combination with exercise, would increase lean body mass and muscle strength in severely burned children more than oxandrolone alone or exercise alone. METHODS AND PATIENTS: Fifty-one burned children (> or = 40% total body surface area burned) were randomly assigned to receive oxandrolone alone (0.1 mg/kg per day orally; n = 9), oxandrolone and exercise (n = 14), placebo and no exercise (n = 11), or placebo and exercise (n = 17). Administration of oxandrolone was started at discharge and continued until 1 year after burn. The 12-week exercise training program was started 6 months after burn. Serum hormones, lean body mass, muscle strength, and peak cardiopulmonary capacity were assessed at 6 (baseline) and 9 months after burn. Data were analyzed using a 1 way analysis of variance, and significance was set at P < .05. RESULTS: The mean percentage of change or increase in weight and lean body mass in the oxandrolone and exercise group was significant compared with placebo and exercise, as well as with the oxandrolone alone group or placebo and no exercise group. Furthermore, lean body mass was significantly improved in the oxandrolone and exercise, oxandrolone alone, and placebo and exercise group compared with the group only receiving placebo. Muscle strength significantly increased in oxandrolone and exercise, placebo and exercise, and the oxandrolone alone group when compared with the placebo and no exercise group. The peak cardiopulmonary capacity was significantly higher in both exercise groups. Insulin-like growth factor 1 was significantly increased in the oxandrolone alone group compared with placebo and exercise and placebo and no exercise. Both exercise groups showed significant changes in insulin-like binding-protein-3 when compared with groups without exercise. CONCLUSIONS: Oxandrolone, in combination with exercise, is beneficial in severely burned children, thus improving their rehabilitation. PMID- 17130282 TI - The molecular mechanisms underlying the reduction of LDL apoB-100 by ezetimibe plus simvastatin. AB - The combination of ezetimibe, an inhibitor of Niemann-Pick C1-like 1 protein (NPC1L1), and an HMG-CoA reductase inhibitor decreases cholesterol absorption and synthesis. In clinical trials, ezetimibe plus simvastatin produces greater LDL cholesterol reductions than does monotherapy. The molecular mechanism for this enhanced efficacy has not been defined. Apolipoprotein B-100 (apoB-100) kinetics were determined in miniature pigs treated with ezetimibe (0.1 mg/kg/day), ezetimibe plus simvastatin (10 mg/kg/day), or placebo (n = 7/group). Ezetimibe decreased cholesterol absorption (-79%) and plasma phytosterols (-91%), which were not affected further by simvastatin. Ezetimibe increased plasma lathosterol (+65%), which was prevented by addition of simvastatin. The combination decreased total cholesterol (-35%) and LDL-cholesterol (-47%). VLDL apoB pool size decreased 26%, due to a 35% decrease in VLDL apoB production. LDL apoB pool size decreased 34% due to an 81% increase in the fractional catabolic rate, both of which were significantly greater than monotherapy. Combination treatment decreased hepatic microsomal cholesterol (-29%) and cholesteryl ester (-65%) and increased LDL receptor (LDLR) expression by 240%. The combination increased NPC1L1 expression in liver and intestine, consistent with increased SREBP2 expression. Ezetimibe plus simvastatin decreases VLDL and LDL apoB-100 concentrations through reduced VLDL production and upregulation of LDLR-mediated LDL clearance. PMID- 17130283 TI - Use of the GRP1 PH domain as a tool to measure the relative levels of PtdIns(3,4,5)P3 through a protein-lipid overlay approach. AB - We describe a novel approach to the relative quantification of phosphatidylinositol (3,4,5)-trisphosphate [PtdIns(3,4,5)P(3)] and its application to measure, in neutrophils, the activation of phosphoinositide 3 kinase (PI3K). This protein-lipid overlay-based assay allowed us to confirm and extend the observations, first, that N-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation of primed human neutrophils leads to a transient and biphasic increase in PtdIns(3,4,5)P(3) levels and, second, that the ability of fMLP to stimulate PtdIns(3,4,5)P(3) accumulation in neutrophils isolated from mice carrying a Ras-insensitive ('DASAA') knock-in of PI3Kgamma (p110gamma(DASAA/DASAA)) is substantially dependent on the Ras binding domain of PI3Kgamma. PMID- 17130284 TI - Checkpoint proteins control morphogenetic events during DNA replication stress in Saccharomyces cerevisiae. AB - In response to DNA replication stress in Saccharomyces cerevisiae, the DNA replication checkpoint maintains replication fork stability, prevents precocious chromosome segregation, and causes cells to arrest as large-budded cells. The checkpoint kinases Mec1 and Rad53 act in this checkpoint. Treatment of mec1 or rad53Delta mutants with replication inhibitors results in replication fork collapse and inappropriate partitioning of partially replicated chromosomes, leading to cell death. We describe a previously unappreciated function of various replication stress checkpoint proteins, including Rad53, in the control of cell morphology. Checkpoint mutants have aberrant cell morphology and cell walls, and show defective bud site selection. Rad53 shows genetic interactions with septin ring pathway components, and, along with other checkpoint proteins, controls the timely degradation of Swe1 during replication stress, thereby facilitating proper bud growth. Thus, checkpoint proteins play an important role in coordinating morphogenetic events with DNA replication during replication stress. PMID- 17130285 TI - An FHA domain-mediated protein interaction network of Rad53 reveals its role in polarized cell growth. AB - The DNA damage checkpoint kinase Rad53 is important for the survival of budding yeast under genotoxic stresses. We performed a biochemical screen to identify proteins with specific affinity for the two Forkhead associated (FHA) domains of Rad53. The N-terminal FHA1 domain was found to coordinate a complex protein interaction network, which includes nuclear proteins involved in DNA damage checkpoints and transcriptional regulation. Unexpectedly, cytosolic proteins involved in cytokinesis, including septins, were also found as FHA1 binding proteins. Consistent with this interaction, a Rad53 mutant defective in its nuclear localization was found to localize to the bud neck. Abnormal morphology was observed in cells overexpressing the FHA1 domain and in rad53Delta cells under DNA replication stress. Further, septin Shs1 appears to have an important role in the response to DNA replication stress. Collectively, the results suggest a novel function of Rad53 in the regulation of polarized cell growth in response to DNA replication stress. PMID- 17130286 TI - Inhibition of Rho A activity causes pemphigus skin blistering. AB - The autoimmune blistering skin diseases pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are mainly caused by autoantibodies against desmosomal cadherins. In this study, we provide evidence that PV-immunoglobulin G (IgG) and PF-IgG induce skin blistering by interference with Rho A signaling. In vitro, pemphigus IgG caused typical hallmarks of pemphigus pathogenesis such as epidermal blistering in human skin, cell dissociation, and loss of desmoglein 1 (Dsg 1) mediated binding probed by laser tweezers. These changes were accompanied by interference with Rho A activation and reduction of Rho A activity. Pemphigus IgG triggered keratinocyte dissociation and Rho A inactivation were p38 mitogen activated protein kinase dependent. Specific activation of Rho A by cytotoxic necrotizing factor-y abolished all pemphigus-triggered effects, including keratin retraction and release of Dsg 3 from the cytoskeleton. These data demonstrate that Rho A is involved in the regulation of desmosomal adhesion, at least in part by maintaining the cytoskeletal anchorage of desmosomal proteins. This may open the possibility of pemphigus treatment with the epidermal application of Rho A agonists. PMID- 17130287 TI - Wnt11 controls cell contact persistence by local accumulation of Frizzled 7 at the plasma membrane. AB - Wnt11 is a key signal, determining cell polarization and migration during vertebrate gastrulation. It is known that Wnt11 functionally interacts with several signaling components, the homologues of which control planar cell polarity in Drosophila melanogaster. Although in D. melanogaster these components are thought to polarize cells by asymmetrically localizing at the plasma membrane, it is not yet clear whether their subcellular localization plays a similarly important role in vertebrates. We show that in zebrafish embryonic cells, Wnt11 locally functions at the plasma membrane by accumulating its receptor, Frizzled 7, on adjacent sites of cell contacts. Wnt11-induced Frizzled 7 accumulations recruit the intracellular Wnt signaling mediator Dishevelled, as well as Wnt11 itself, and locally increase cell contact persistence. This increase in cell contact persistence is mediated by the local interaction of Wnt11, Frizzled 7, and the atypical cadherin Flamingo at the plasma membrane, and it does not require the activity of further downstream effectors of Wnt11 signaling, such as RhoA and Rok2. We propose that Wnt11, by interacting with Frizzled 7 and Flamingo, modulates local cell contact persistence to coordinate cell movements during gastrulation. PMID- 17130288 TI - Did2 coordinates Vps4-mediated dissociation of ESCRT-III from endosomes. AB - The sorting of transmembrane cargo proteins into the lumenal vesicles of multivesicular bodies (MVBs) depends on the recruitment of endosomal sorting complexes required for transport (ESCRTs) to the cytosolic face of endosomal membranes. The subsequent dissociation of ESCRT complexes from endosomes requires Vps4, a member of the AAA family of adenosine triphosphatases. We show that Did2 directs Vps4 activity to the dissociation of ESCRT-III but has no role in the dissociation of ESCRT-I or -II. Surprisingly, vesicle budding into the endosome lumen occurs in the absence of Did2 function even though Did2 is required for the efficient sorting of MVB cargo proteins into lumenal vesicles. This uncoupling of MVB cargo sorting and lumenal vesicle formation suggests that the Vps4-mediated dissociation of ESCRT-III is an essential step in the sorting of cargo proteins into MVB vesicles but is not a prerequisite for the budding of vesicles into the endosome lumen. PMID- 17130289 TI - Human SHPRH suppresses genomic instability through proliferating cell nuclear antigen polyubiquitination. AB - Differential modifications of proliferating cell nuclear antigen (PCNA) determine DNA repair pathways at stalled replication forks. In yeast, PCNA monoubiquitination by the ubiquitin ligase (E3) yRad18 promotes translesion synthesis (TLS), whereas the lysine-63-linked polyubiquitination of PCNA by yRad5 (E3) promotes the error-free mode of bypass. The yRad5-dependent pathway is important to prevent genomic instability during replication, although its exact molecular mechanism is poorly understood. This mechanism has remained totally elusive in mammals because of the lack of apparent RAD5 homologues. We report that a putative tumor suppressor gene, SHPRH, is a human orthologue of yeast RAD5. SHPRH associates with PCNA, RAD18, and the ubiquitin-conjugating enzyme UBC13 (E2) and promotes methyl methanesulfonate (MMS)-induced PCNA polyubiquitination. The reduction of SHPRH by stable short hairpin RNA increases sensitivity to MMS and enhances genomic instability. Therefore, the yRad5/SHPRH dependent pathway is a conserved and fundamental DNA repair mechanism that protects the genome from genotoxic stress. PMID- 17130290 TI - Requirement of biphasic calcium release from the endoplasmic reticulum for Fas mediated apoptosis. AB - Fas receptor is a member of the tumor necrosis factor-alpha family of death receptors that mediate physiologic apoptotic signaling. To investigate the molecular mechanisms regulating calcium mobilization during Fas-mediated apoptosis, we have analyzed the sequential steps leading to altered calcium homeostasis and cell death in response to activation of the Fas receptor. We show that Fas-mediated apoptosis requires endoplasmic reticulum-mediated calcium release in a mechanism dependent on phospholipase C-gamma1 (PLC-gamma1) activation and Ca2+ release from inositol 1,4,5-trisphosphate receptor (IP3R) channels. The kinetics of Ca2+ release were biphasic, demonstrating a rapid elevation caused by PLC-gamma1 activation and a delayed and sustained increase caused by cytochrome c binding to IP3R. Blocking either phase of Ca2+ mobilization was cytoprotective, highlighting PLC-gamma1 and IP3R as possible therapeutic targets for disorders associated with Fas signaling. PMID- 17130291 TI - Unassisted translocation of large polypeptide domains across phospholipid bilayers. AB - Although transmembrane proteins generally require membrane-embedded machinery for integration, a few can insert spontaneously into liposomes. Previously, we established that the tail-anchored (TA) protein cytochrome b(5) (b5) can posttranslationally translocate 28 residues downstream to its transmembrane domain (TMD) across protein-free bilayers (Brambillasca, S., M. Yabal, P. Soffientini, S. Stefanovic, M. Makarow, R.S. Hegde, and N. Borgese. 2005. EMBO J. 24:2533-2542). In the present study, we investigated the limits of this unassisted translocation and report that surprisingly long (85 residues) domains of different sequence and charge placed downstream of b5's TMD can posttranslationally translocate into mammalian microsomes and liposomes at nanomolar nucleotide concentrations. Furthermore, integration of these constructs occurred in vivo in translocon-defective yeast strains. Unassisted translocation was not unique to b5 but was also observed for another TA protein (protein tyrosine phosphatase 1B) whose TMD, like the one of b5, is only moderately hydrophobic. In contrast, more hydrophobic TMDs, like synaptobrevin's, were incapable of supporting unassisted integration, possibly because of their tendency to aggregate in aqueous solution. Our data resolve long-standing discrepancies on TA protein insertion and are relevant to membrane evolution, biogenesis, and physiology. PMID- 17130292 TI - Circadian oscillators of Drosophila and mammals. PMID- 17130293 TI - Differentiation of columnar epithelia: the hensin pathway. AB - Epithelia, the most common variety of cells in complex organisms exist in many shapes. They are sheets of polarized cells that separate two compartments and selectively transport materials from one to the other. After acquiring these general characteristics, they differentiate to become specialized types such as squamous columnar or transitional epithelia. High density seeding converts a kidney-derived cell line from flat ;generic' epithelial cells to columnar cells. The cells acquire all the characteristics of differentiated columnar cells, including microvilli, and the capacity for apical endocytosis. The high seeding density induces the deposition of a new protein termed hensin and polymerization of hensin is the crucial event that dictates changes in epithelial phenotype. Hensin is widely expressed in most epithelia. Its deletion in mice leads to embryonic lethality at the time of generation of the first columnar epithelium, the visceral endoderm. Moreover many human cancers have deletions in the hensin gene, which indicates that it is a tumor suppressor. PMID- 17130294 TI - All in the CCN family: essential matricellular signaling modulators emerge from the bunker. AB - The CCN family is a group of six secreted proteins that specifically associate with the extracellular matrix. Structurally, CCN proteins are modular, containing up to four distinct functional domains. CCN family members are induced by growth factors and cytokines such as TGFbeta and endothelin 1 and cellular stress such as hypoxia, and are overexpressed in pathological conditions that affect connective tissues, including scarring, fibrosis and cancer. Although CCN family members were discovered over a decade ago, the precise biological role, mechanism of action and physiological function of these proteins has remained elusive until recently, when several key mechanistic insights into the CCN family emerged. The CCNs have been shown to have key roles as matricellular proteins, serving as adaptor molecules connecting the cell surface and extracellular matrix (ECM). Although they appear not to have specific high-affinity receptors, they signal through integrins and proteoglycans. Furthermore, in addition to having inherent adhesive abilities that modulate focal adhesions and control cell attachment and migration, they execute their functions by modulating the activity of a variety of different growth factors, such as TGFbeta. CCN proteins not only regulate crucial biological processes including cell differentiation, proliferation, adhesion, migration, apoptosis, ECM production, chondrogenesis and angiogenesis, but also have more sinister roles promoting conditions such as fibrogenesis. PMID- 17130295 TI - Distinct subdomain organization and molecular composition of a tight junction with adherens junction features. AB - Most polarized epithelia constrain solute diffusion between luminal and interstitial compartments using tight junctions and generate mechanical strength using adherens junctions. These intercellular junctions are typically portrayed as incongruent macromolecular complexes with distinct protein components. Herein, we delineate the molecular composition and subdomain architecture of an intercellular junction between sensory and non-sensory cells of the inner ear. In this junction, claudins partition into claudin-14 and claudin-9/6 subdomains that are distinguishable by strand morphology, which contrasts with in vitro data that most claudins co-assemble into heteromeric strands. Surprisingly, canonical adherens junction proteins (p120ctn, alpha- and beta-catenins) colocalize with the claudin-9/6 subdomain and recruit a dense cytoskeletal network. We also find that catenins colocalize with claudin-9 and claudin-6, but not claudin-14, in a heterologous system. Together, our data demonstrate that canonical tight junction and adherens junction proteins can be recruited to a single junction in which claudins partition into subdomains and form a novel hybrid tight junction with adherens junction organization. PMID- 17130296 TI - Putative role of beta-1,3 glucans in Candida albicans biofilm resistance. AB - Biofilms are microbial communities, embedded in a polymeric matrix, growing attached to a surface. Nearly all device-associated infections involve growth in the biofilm life style. Biofilm communities have characteristic architecture and distinct phenotypic properties. The most clinically important phenotype involves extraordinary resistance to antimicrobial therapy, making biofilm infections very difficulty to cure without device removal. The current studies examine drug resistance in Candida albicans biofilms. Similar to previous reports, we observed marked fluconazole and amphotericin B resistance in a C. albicans biofilm both in vitro and in vivo. We identified biofilm-associated cell wall architectural changes and increased beta-1,3 glucan content in C. albicans cell walls from a biofilm compared to planktonic organisms. Elevated beta-1,3 glucan levels were also found in the surrounding biofilm milieu and as part of the matrix both from in vitro and in vivo biofilm models. We thus investigated the possible contribution of beta-glucans to antimicrobial resistance in Candida albicans biofilms. Initial studies examined the ability of cell wall and cell supernatant from biofilm and planktonic C. albicans to bind fluconazole. The cell walls from both environmental conditions bound fluconazole; however, four- to fivefold more compound was bound to the biofilm cell walls. Culture supernatant from the biofilm, but not planktonic cells, bound a measurable amount of this antifungal agent. We next investigated the effect of enzymatic modification of beta-1,3 glucans on biofilm cell viability and the susceptibility of biofilm cells to fluconazole and amphotericin B. We observed a dose-dependent killing of in vitro biofilm cells in the presence of three different beta-glucanase preparations. These same concentrations had no impact on planktonic cell viability. beta-1,3 Glucanase markedly enhanced the activity of both fluconazole and amphotericin B. These observations were corroborated with an in vivo biofilm model. Exogenous biofilm matrix and commercial beta-1,3 glucan reduced the activity of fluconazole against planktonic C. albicans in vitro. In sum, the current investigation identified glucan changes associated with C. albicans biofilm cells, demonstrated preferential binding of these biofilm cell components to antifungals, and showed a positive impact of the modification of biofilm beta-1,3 glucans on drug susceptibility. These results provide indirect evidence suggesting a role for glucans in biofilm resistance and present a strong rationale for further molecular dissection of this resistance mechanism to identify new drug targets to treat biofilm infections. PMID- 17130297 TI - Antiviral activities of novel 5-phosphono-pent-2-en-1-yl nucleosides and their alkoxyalkyl phosphonoesters. AB - Three acyclic nucleoside phosphonates are currently approved for clinical use against infections caused by cytomegalovirus (Vistide), hepatitis B virus (Hepsera), and human immunodeficiency virus type 1 (Viread). This important antiviral class inhibits viral polymerases after cellular uptake and conversion to their diphosphates, bypassing the first phosphorylation, which is required for conventional nucleoside antivirals. Small chemical alterations in the acyclic side chain lead to marked differences in antiviral activity and the spectrum of activity of acyclic nucleoside phosphonates against various classes of viral agents. We synthesized a new class of acyclic nucleoside phosphonates based on a 5-phosphono-pent-2-en-1-yl base motif in which the oxygen heteroatom usually present in acyclic nucleoside phosphonates has been replaced with a double bond. Since the intrinsic phosphonate moiety leads to low oral bioavailability and impaired cellular penetration, we also prepared the hexadecyloxypropyl esters of the 5-phosphono-pent-2-en-1-yl nucleosides. Our earlier work showed that this markedly increases antiviral activity and oral bioavailability. Although the 5 phosphono-pent-2-en-1-yl nucleosides themselves were not active, the hexadecyloxypropyl esters were active against DNA viruses and hepatitis B virus, in vitro. Notably, the hexadecyloxypropyl ester of 9-(5-phosphono-pent-2-en-1-yl) adenine was active against hepatitis B virus mutants resistant to lamivudine, emtricitabine, and adefovir. PMID- 17130298 TI - Vancomycin-intermediate Staphylococcus aureus strains have impaired acetate catabolism: implications for polysaccharide intercellular adhesin synthesis and autolysis. AB - The most common mechanism by which Staphylococcus aureus gains resistance to vancomycin is by adapting its physiology and metabolism to permit growth in the presence of vancomycin. Several studies have examined the adaptive changes occurring during the transition to vancomycin-intermediate resistance, leading to a model of vancomycin resistance in which decreased cell wall turnover and autolysis result in increased cell wall thickness and resistance to vancomycin. In the present study, we identified metabolic changes common to vancomycin intermediate S. aureus (VISA) strains by assessing the metabolic and growth characteristics of two VISA strains (vancomycin MICs of 8 microg/ml) and two isogenic derivative strains with vancomycin MICs of 32 microg/ml. Interestingly, we observed the parental strains had impaired catabolism of nonpreferred carbon sources (i.e., acetate), and this impairment became more pronounced as vancomycin resistance increased. To determine if acetate catabolism impairment is common to VISA strains, we assessed the ability of VISA and vancomycin-sensitive S. aureus (VSSA) clinical isolates to catabolize acetate. As expected, a significantly greater percentage of VISA strains (71%) had impaired acetate catabolism relative to VSSA (8%). This is an important observation because staphylococcal acetate catabolism is implicated in growth yield and antibiotic tolerance and in regulating cell death and polysaccharide intercellular adhesin synthesis. PMID- 17130299 TI - Deregulated Syk inhibits differentiation and induces growth factor-independent proliferation of pre-B cells. AB - The nonreceptor protein spleen tyrosine kinase (Syk) is a key mediator of signal transduction in a variety of cell types, including B lymphocytes. We show that deregulated Syk activity allows growth factor-independent proliferation and transforms bone marrow-derived pre-B cells that are then able to induce leukemia in mice. Syk-transformed pre-B cells show a characteristic pattern of tyrosine phosphorylation, increased c-Myc expression, and defective differentiation. Treatment of Syk-transformed pre-B cells with a novel Syk-specific inhibitor (R406) reduces tyrosine phosphorylation and c-Myc expression. In addition, R406 treatment removes the developmental block and allows the differentiation of the Syk-transformed pre-B cells into immature B cells. Because R406 treatment also prevents the proliferation of c-Myc-transformed pre-B cells, our data indicate that endogenous Syk kinase activity may be required for the survival of pre-B cells transformed by other oncogenes. Collectively, our data suggest that Syk is a protooncogene involved in the transformation of lymphocytes, thus making Syk a potential target for the treatment of leukemia. PMID- 17130300 TI - Role of IL-17 and regulatory T lymphocytes in a systemic autoimmune disease. AB - To explore the interactions between regulatory T cells and pathogenic effector cytokines, we have developed a model of a T cell-mediated systemic autoimmune disorder resembling graft-versus-host disease. The cytokine responsible for tissue inflammation in this disorder is interleukin (IL)-17, whereas interferon (IFN)-gamma produced by Th1 cells has a protective effect in this setting. Because of the interest in potential therapeutic approaches utilizing transfer of regulatory T cells and inhibition of the IL-2 pathway, we have explored the roles of these in the systemic disease. We demonstrate that the production of IL-17 and tissue infiltration by IL-17-producing cells occur and are even enhanced in the absence of IL-2. Regulatory T cells favor IL-17 production but prevent the disease when administered early in the course by suppressing expansion of T cells. Thus, the pathogenic or protective effects of cytokines and the therapeutic capacity of regulatory T cells are crucially dependent on the timing and the nature of the disease. PMID- 17130301 TI - Screening random peptide libraries with subacute sclerosing panencephalitis brain derived recombinant antibodies identifies multiple epitopes in the C-terminal region of the measles virus nucleocapsid protein. AB - Infectious and inflammatory diseases of the CNS are often characterized by a robust B-cell response that manifests as increased intrathecal immunoglobulin G (IgG) synthesis and the presence of oligoclonal bands. We previously used laser capture microdissection and single-cell PCR to analyze the IgG variable regions of plasma cells from the brain of a patient with subacute sclerosing panencephalitis (SSPE). Five of eight human IgG1 recombinant antibodies (rAbs) derived from SSPE brain plasma cell clones recognized the measles virus (MV) nucleocapsid protein, confirming that the antibody response in SSPE targets primarily the agent causing disease. In this study, as part of our work on antigen identification, we used four rAbs to probe a random phage-displayed peptide library to determine if epitopes within the MV nucleocapsid protein could be identified with SSPE brain rAbs. All four of the SSPE rAbs enriched phage displayed peptide sequences that reacted specifically to their panning rAb by enzyme-linked immunosorbent assay. BLASTP searches of the NCBI protein database revealed clear homologies in three peptides and different amino acid stretches within the 65 C-terminal amino acids of the MV nucleocapsid protein. The specificities of SSPE rAbs to these regions of the MV nucleocapsid protein were confirmed by binding to synthetic peptides or to short cDNA expression products. These results indicate the feasibility of using peptide screening for antigen discovery in central nervous system inflammatory diseases of unknown etiology, such as multiple sclerosis, neurosarcoidosis, or Behcet's syndrome. PMID- 17130302 TI - Expression of a sorcin missense mutation in the heart modulates excitation contraction coupling. AB - Sorcin is a Ca2+ binding protein implicated in the regulation of intracellular Ca2+ cycling and cardiac excitation-contraction coupling. Structural and human genetic studies suggest that a naturally occurring sequence variant encoding L112 sorcin disrupts an E-F hand Ca2+ binding domain and may be responsible for a heritable form of hypertension and hypertrophic heart disease. We generated transgenic mice overexpressing L112-sorcin in the heart and characterized the effects on Ca2+ regulation and cardiac function both in vivo and in dissociated cardiomyocytes. Hearts of sorcin(F112L) transgenic mice were mildly dilated but ventricular function was preserved and systemic blood pressure was normal. Sorcin(F112L) myocytes were smaller than control cells and displayed complex alterations in Ca2+ regulation and contractility, including a slowed inactivation of L-type Ca2+ current, enhanced Ca2+ spark width, duration, and frequency, and increased Na+-Ca2+ exchange activity. In contrast, mice with cardiac-specific overexpression of wild-type sorcin displayed directionally opposite effects on L type Ca2+ channel function and Ca2+ spark behavior. These data further define the role of sorcin in cardiac excitation-contraction coupling and highlight its negative regulation of SR calcium release. Our results also suggest that additional factors may be responsible for the development of cardiac hypertrophy and hypertension in humans expressing the L112-sorcin sequence variant. PMID- 17130303 TI - Relationship between low-normal blood pressure and kidney disease in type 1 diabetes. AB - Current recommendations, largely based on studies in type 2 diabetes, suggest lower target blood pressures (BPs) for individuals with diabetes than for the general population. However, the effect of lower BP on renal outcomes in type 1 diabetes is uncertain. In a population-based cohort of type 1 diabetes adults (mean age: 33.1 years) based in Wisconsin, of which the distribution of baseline BP was in the low-normal range, we examined the relationship between decreasing categories of systolic and diastolic BP and the 16-year incidence of proteinuria (n=232 of 604) and estimated glomerular filtration rate of <60 mL/min/1.73 m(2) (n=158 of 547). Decreasing BP categories had lower relative risk (RR) of developing incident proteinuria (RR comparing decreasing quartiles of systolic BP: 1.00, 0.76, 0.58, 0.73; P for trend=0.03; RR comparing decreasing quartiles of diastolic BP: 1.00, 0.81, 0.66, 0.42; P for trend <0.0001) and incident estimated glomerular filtration rate <60 mL/min/1.73 m(2) (RR comparing decreasing quartiles of systolic BP: 1.00, 0.83, 0.61, 0.65; P for trend=0.03; RR comparing decreasing quartiles of diastolic BP: 1.00, 0.84, 0.82, 0.43; P for trend=0.001). These associations were independent of glycemic control and several putative confounding factors. Subjects with either systolic BP <120 mm Hg or diastolic BP <70 mm Hg had significantly lower RR (95% confidence interval) of incident proteinuria (0.63 [0.48 to 0.82]) and incident estimated glomerular filtration rate <60 mL/min/1.73 m(2) (0.60 [0.43 to 0.82]); corresponding population-attributable risks for these outcomes were 26.7% and 29.5%, respectively. Our study suggests that lower BP levels, even below the accepted normal range, are protective against kidney disease in adults with type 1 diabetes. Interventional trials are desirable to clarify the clinical significance of this association. PMID- 17130304 TI - Contribution of endothelial nitric oxide to blood pressure in humans. AB - Impaired endothelial-derived NO (eNO) is invoked in the development of many pathological conditions. Systemic inhibition of NO synthesis, used to assess the importance of NO to blood pressure (BP) regulation, increases BP by approximately 15 mm Hg. This approach underestimates the importance of eNO, because BP is restrained by baroreflex mechanisms and does not account for a role of neurally derived NO. To overcome these limitations, we induced complete autonomic blockade with trimethaphan in 17 normotensive healthy control subjects to eliminate baroreflex mechanisms and contribution of neurally derived NO. Under these conditions, the increase in BP reflects mostly blockade of tonic eNO. N(G) Monomethyl-l-arginine (250 microg/kg per minute IV) increased mean BP by 6+/-3.7 mm Hg (from 77 to 82 mm Hg) in intact subjects and by 21+/-8.4 mm Hg (from 75 to 96 mm Hg) during autonomic blockade. We did not find a significant contribution of neurally derived NO to BP regulation after accounting for baroreflex buffering. To further validate this approach, we compared the effect of NOS inhibition during autonomic blockade in 10 normotensive individuals with that of 6 normotensive smokers known to have endothelial dysfunction but who were otherwise normal. As expected, normotensive smokers showed a significantly lower increase in systolic BP during selective eNO blockade (11+/-4.5 versus 30+/-2.3 mm Hg in normotensive individuals; P<0.005). Thus, we report a novel approach to preferentially evaluate the role of eNO on BP control in normal and disease states. Our results suggest that eNO is one of the most potent metabolic determinants of BP in humans, tonically restraining it by approximately 30 mm Hg. PMID- 17130305 TI - How do you define "hypertension" in a patient with type 1 diabetes? PMID- 17130306 TI - Effect of tryptophan hydroxylase 1 deficiency on the development of hypoxia induced pulmonary hypertension. AB - Tryptophan hydroxylase 1 catalyzes the rate-limiting step in the synthesis of serotonin in the periphery. Recently, it has been shown that expression of the tryptophan hydroxylase 1 gene is increased in lungs and pulmonary endothelial cells from patients with idiopathic pulmonary arterial hypertension. Here we investigated the effect of genetic deletion of tryptophan hydroxylase 1 on hypoxia-induced pulmonary arterial hypertension in mice by measuring pulmonary hemodynamics and pulmonary vascular remodeling before and after 2 weeks of hypoxia. In wild-type mice, hypoxia increased right ventricular pressure and pulmonary vascular remodeling. These effects of hypoxia were attenuated in the tryptophan hydroxylase 1-/-mice. Hypoxia increased right ventricular hypertrophy in both wild-type and tryptophan hydroxylase 1-/-mice suggesting that in vivo peripheral serotonin has a differential effect on the pulmonary vasculature and right ventricular hypertrophy. Contractile responses to serotonin were increased in pulmonary arteries from tryptophan hydroxylase 1-/-mice. Hypoxia increased serotonin-mediated contraction in vessels from the wild-type mice, but this was not further increased by hypoxia in the tryptophan hydroxylase 1-/-mice. In conclusion, these results indicate that tryptophan hydroxylase 1 and peripheral serotonin play an essential role in the development of hypoxia-induced elevations in pulmonary pressures and hypoxia-induced pulmonary vascular remodeling. In addition, the results suggest that, in mice, serotonin has differential effects on the pulmonary vasculature and right ventricular hypertrophy. PMID- 17130307 TI - Metabolic syndrome and early death: getting to the heart of the problem. PMID- 17130308 TI - Metabolic syndrome in the Pressioni Arteriose Monitorate E Loro Associazioni (PAMELA) study: daily life blood pressure, cardiac damage, and prognosis. AB - The prevalence of the metabolic syndrome (National Cholesterol Education Program Adult Treatment Panel III criteria) and its relationships with daily life blood pressures, cardiac damage, and prognosis were determined in 2013 subjects from a Northern Italian population aged 25 to 74 years. Home blood pressure, 24-hour blood pressure, and left ventricular mass index (echocardiography) were also measured. Cardiovascular and noncardiovascular deaths were registered over 148 months. Metabolic syndrome was found in 16.2% of the sample, an office blood pressure elevation being the most frequent (95.4%) and the blood glucose abnormality the least frequent (31.5%) component. There was in metabolic syndrome a frequent elevation in home and/or 24-hour average blood pressure, as well as a greater left ventricular mass index and prevalence of left ventricular hypertrophy, which was manifest even when data were adjusted for between-group differences, including blood pressure. The adjusted risk of cardiovascular and all-cause mortality was greater in metabolic syndrome subjects (+71.0% and +37.0%; P<0.05), a further marked increase being observed with left ventricular hypertrophy or "in-office" and "out-of-office" blood pressure elevations. The increased risk was related to the blood pressure and the blood glucose component of metabolic syndrome, with no contribution of the remaining components. Thus, metabolic syndrome is common in a Mediterranean population in which it significantly increases the long-term risk of death. Cardiac abnormalities and increases in home and 24-hour blood pressure are common in metabolic syndrome, and their occurrence further enhances the risk. The contribution of metabolic syndrome components to the risk, however, is unbalanced and mainly related to blood pressure and glucose abnormalities. PMID- 17130309 TI - Morbid obesity and left ventricular geometry. PMID- 17130310 TI - Left ventricular hypertrophy in severe obesity: interactions among blood pressure, nocturnal hypoxemia, and body mass. AB - Obese subjects have a high prevalence of left ventricular (LV) hypertrophy. It is unclear to what extent LV hypertrophy results directly from obesity or from associated conditions, such as hypertension, impaired glucose homeostasis, or obstructive sleep apnea. We tested the hypothesis that LV hypertrophy in severe obesity is associated with additive effects from each of the major comorbidities. Echocardiography and laboratory testing were performed in 455 severely obese subjects with body mass index 35 to 92 kg/m(2) and 59 nonobese reference subjects. LV hypertrophy, defined by allometrically corrected (LV mass/height(2.7)), gender-specific criteria, was present in 78% of the obese subjects. Multivariable regression analyses showed that average nocturnal oxygen saturation <85% was the strongest independent predictor of LV hypertrophy (P<0.001), followed by systolic blood pressure (P<0.015) and then body mass index (P<0.05). With regard to LV mass, there were synergistic effects between hypertension and body mass index (P interaction <0.001) and between hypertension and reduced nocturnal oxygen saturation. Severely obese subjects had normal LV endocardial fractional shortening (35+/-6% versus 35+/-6%) but mildly decreased midwall fractional shortening (15+/-2% versus 17+/-2%; P<0.001), indicating subtle myocardial dysfunction. In conclusion, more severe nocturnal hypoxemia, increasing systolic blood pressure, and body mass index are all independently associated with increased LV mass. The effects of increased blood pressure seem to amplify those of sleep apnea and more severe obesity. PMID- 17130311 TI - Treatment methods for spinal cord injury caused by acute type B aortic dissection. AB - Acute distal aortic dissection rarely causes spinal cord ischemia presenting with paraplegia or paraparesis. Spinal cord involvement has poor outcomes, and there is no established effective treatment for this disorder. In this report we describe the acute conservative treatment of two cases of paraplegia/paraparesis due to acute type B aortic dissection. Early reversal of lower-limb dysmobility was achieved. PMID- 17130312 TI - Thoracoscopic thymectomy in a previous sternotomy. AB - A video assisted thoracoscopic surgery (VATS) thymectomy is a viable alternative to conventional open thymectomy in myasthenia gravis (MG). A previous operation in the same anatomical region is generally considered a relative contraindication to any minimally invasive approach in the same area. Few cases of VATS on previously operated chests have been reported. We report a case of a VATS thymectomy in a patient who had undergone two previous sternotomies for cardiac disease. PMID- 17130313 TI - Acute fulminant myocarditis after diphtheria, polio, and tetanus vaccination. AB - We report an infant case of acute fulminant myocarditis which occurred after administration of a diphtheria, polio, and tetanus vaccination. Fever and dyspnea developed after the vaccination. Extracorporeal membrane oxygenation was used for intractable cardiogenic shock. The patient survived the extracorporeal support, but poor ventricular contractility recurred 2 months later and she died while waiting for heart transplantation. PMID- 17130314 TI - Pulmonary oncocytoma: a rare case. AB - Oncocytomas are rare tumors with characteristic histological features of large eosinophilic cells with prominent nucleoli and small round nuclei. They commonly occur in the kidney and salivary glands. Their occurrence in the lungs is very rare; fewer than 6 cases being reported so far in the available world literature. We encountered one such case and present the details of the management and its outcome. PMID- 17130315 TI - Infective endocarditis associated with Sjogren's syndrome. AB - Sjogren's syndrome is one of the major autoimmune diseases, however infective endocarditis associated with Sjogren's syndrome has not previously been reported. A patient with Sjogren's syndrome associated with aortic valve regurgitation due to infective endocarditis, underwent successful aortic valve replacement. Patients with Sjogren's syndrome tend to be at a higher risk of intraoral infections due to diminished secretion of saliva. Particular care must be taken of Sjogren's syndrome patients as they can also be at an increased risk of infective endocarditis. PMID- 17130316 TI - Totally endoscopic coronary artery bypass surgery. AB - Robotically enhanced surgery is a fast-developing technique that allows totally endoscopic cardiac surgery on both the beating and arrested heart. Between December 2002 and May 2005, 13 patients underwent totally endoscopic coronary bypass using the da Vinci system; 11 operations were on a beating heart and 2 on arrested hearts. The mean time for internal mammary artery mobilization was 42 min. The time for left internal mammary artery-to-left anterior descending artery anastomosis was 20-36 min for totally endoscopic cases. In one patient, the right internal mammary artery was anastomosed to the diagonal artery. No patient required conversion to a median sternotomy. Mean intensive care unit stay was 1.2 days and mean hospital stay was 4.5 days. There was no hospital mortality. All 13 patients had coronary angiography at 3-month intervals, which showed 100% patency in 12 patients while one had 50% anastomotic narrowing for which coronary angioplasty was performed. Using robotic technology, completely endoscopic anastomosis is possible in patients with single-vessel disease. Use of robotics is now extended to achieve complete myocardial revascularization by harvesting both internal mammary arteries in addition to making a small thoracotomy for direct anastomosis. PMID- 17130317 TI - Pneumatic pump reduces leg wound complications in cardiac patients. AB - Leg wound complications at the site of vein harvest for coronary artery bypass graft, although infrequent, cause significant morbidity. Pneumatic pressure therapy is valuable in venous and lymphatic diseases, but its usefulness after leg vein harvest has not been determined. A prospective randomized controlled trial was conducted on 200 patients, half of whom had sequential pneumatic leg pump therapy postoperatively. Wound healing, extent of lower limb edema, patient satisfaction, and the financial implications of pneumatic pressure therapy were assessed. In the study group, 71 patients had satisfactory wound healing vs. 23 in the control group. The leg wound infection rate in the study group was 3% vs. 15% in the control group ( p = 0.003). Lower limb edema was significantly reduced in the study group in the early postoperative period ( p < 0.05), and the mean postoperative length of hospital stay was reduced by 2.6 days in patients given pneumatic pressure therapy ( p = 0.003). The sequential pneumatic leg pump is an effective, inexpensive, and convenient device that reduces leg wound complications after coronary artery bypass grafting. PMID- 17130318 TI - Long-term outcome of intracardiac repair under simple deep hypothermia. AB - Development of cardiopulmonary bypass has contributed to pediatric cardiac surgery, but at the dawn of cardiac surgery, simple deep hypothermia was used to avoid the deleterious effect of cardiopulmonary bypass. Between 1981 and 1990, 45 patients with simple cardiac anomalies underwent definitive surgery under deep hypothermia. Age at operation was 35 days to 20 months, and body weight was 2.3 to 8.0 kg. Under morphine and ether anesthesia, a median sternotomy was performed when the esophageal temperature reached 26.3 degrees C +/- 1.3 degrees C by the application of surface cooling. At a minimum esophageal temperature of 19.6 degrees C +/- 2.3 degrees C, inflow occlusion and cold cardioplegia were applied to induce circulatory arrest for 32.4 +/- 10.2 min. Direct cardiac massage was used to restore cardiac activity during rewarming. All but one patient was in New York Heart Association functional class I postoperatively. The latest cardiothoracic ratio was 49.8% +/- 4.7%. All but 2 patients are free from medication. Five of 30 patients showed developmental delay in the long-term; 2 of these had a long circulatory arrest period, and 3 had prolonged heart failure postoperatively. The other 25 patients had excellent physiologic and mental development. The long-term outcome of perfusionless hypothermic cardiac surgery is satisfactory when applied appropriately. PMID- 17130319 TI - Mortality and morbidity after aortic root replacement: 10-year experience. AB - Aortic root reconstruction remains a challenging surgical procedure. This retrospective study was carried out to evaluate the early and long-term outcomes of aortic root replacement over a 10-year period. There were 83 patients with a mean age of 43.2 +/- 14 years (range, 10 to 78 years). Type A aortic dissection and Marfan syndrome were found in 28% and 24%, respectively. The most common technique used for repair of this condition was the Bentall operation. The mean duration of follow-up was 29.6 +/- 28 months, ranging from 1 to 120 months. Hospital (30-day) mortality was 13.3% (11 patients). Two patients died during the late follow-up. The mortality was significantly higher in patients presenting with cardiogenic shock, those with long cardiopulmonary bypass and crossclamp times, and the group who had concomitant coronary artery bypass grafting. Emergency operation was not a significant risk factor for early death in our patients. The most common complications were bleeding and neurological sequelae. Aortic root replacement can be achieved with acceptable mortality and morbidity in a high-risk group of patients. Improvements in the outcome may be achieved by faster transport of patients in cardiogenic shock, and by reducing the cardiopulmonary bypass and crossclamp times. PMID- 17130320 TI - Late re-operation for aortic and mitral Starr-Edwards ball valve prostheses. AB - Starr-Edwards ball valves removed more than 15 years after implantation were retrospectively investigated macroscopically. Eight patients required re operation. Valve models used in the initial operations were a non-cloth-covered valve in 2 patients and a cloth-covered valve in 6. Two patients had replacement of an aortic ball valve (model 1260 and model 2320) and 6 underwent mitral valve replacement (model 6120 in one, model 6320 in 5). The mean time to re-operation was 23.0 +/- 4.8 years after implantation. Cloth wear causing significant hemolysis was observed in all cloth-covered valves, regardless of valve position. Autologous tissue growth was noted on the orifice ring and struts in both aortic and mitral prostheses. Thrombus formation was not found in any of the valves. Ball variance in silicone rubber balls was mild in the non-cloth-covered valves, even in the aortic position. The most significant problem with the cloth-covered ball valve was cloth wear. Cloth wear should always be considered when 15 years or more have passed since valve implantation. Significant hemolysis, elevation of lactate dehydrogenase values, and echocardiographic detection of transvalvular regurgitation are diagnostic of cloth wear, and are indications for replacement of a cloth-covered ball valve. PMID- 17130321 TI - Modification of thoracoscopic surgery for spontaneous pneumothorax. AB - Morbidity, use of analgesics, postoperative drainage, and hospital stay are reduced after video-assisted thoracoscopic surgery for pneumothorax. However, some surgeons prefer a minithoracotomy because the rate of recurrence after thoracoscopic surgery is 5%-10%. A modified thoracoscopic bullectomy is described, which has the advantages of both conventional video-assisted thoracoscopic surgery and a minithoracotomy. Of 69 patients who underwent surgery for pneumothorax from January 2002 to February 2003, 13 were treated by conventional video-assisted thoracoscopic surgery and 21 by the modified thoracoscopic bullectomy. The mean ages were 20.6 years in the conventional group and 23.0 years in the modified group, with follow-up of 25.8 +/- 1.8 months in the conventional group and 20.6 +/- 1.3 months in the modified group. The duration of operation was similar in both groups (49.3 +/- 16.0 vs. 44.2 +/- 19.2 min). Significantly fewer staples were used in the modified group (1.62 +/- 0.74 vs. 2.92 +/- 1.19). The duration of chest tube drainage and postoperative hospital stay were significantly reduced in the modified group. The modified thoracoscopic bullectomy is an effective procedure for the treatment of primary spontaneous pneumothorax. PMID- 17130322 TI - Asymptomatic solitary mediastinal cystic lymphangioma: a rare entity. AB - Solitary mediastinal cystic lymphangioma is an extremely rare tumor-like abnormality of the lymphatic system, of congenital origin. From 1996 to 2003, 4 cases of solitary mediastinal cystic lymphangioma were managed in our department. The clinicopathologic characteristics were recorded and analyzed. The incidence proved to be 1.2% among all cases of mediastinal masses admitted to our department. All patients were asymptomatic, they shared common radiologic features, and all were managed by radical surgical excision. No recurrence or mortality were noted during follow-up ranging from 2 to 7 years. Complete excision of this benign entity has an excellent prognosis. PMID- 17130323 TI - Coronary artery bypass surgery in the elderly: experience of tertiary care hospital. AB - In the subcontinent, improving life expectancy may increase the number of elderly patients requiring coronary artery bypass. The elderly have a higher risk of mortality, and cost vs benefit is presumed to be unrewarding, leading to a disinclination to operate on such patients. We reviewed the records of patients >or= 70 years old undergoing coronary revascularization in a 10-year period. Between 1995 and 2004, 3,312 coronary bypass operations were performed, including 242 (7.3%) in patients >or= 70 years. Of these, 207 (86%) were male, 73 (30%) had non-elective surgery, and 177 (73%) had a high-risk EuroSCORE. Early postoperative mortality was 8.7% (21/242), and 33 (13.6%) patients experienced morbidity. One-year follow-up of 106 (44%) patients was possible. There were 4 (1.7%) late deaths from noncardiac causes. Death, morbidity, and postoperative functional class were significantly associated with preoperative functional class. Meticulous selection of elderly patients for coronary artery bypass may help maximize the benefits of this surgery. PMID- 17130324 TI - Postcardiotomy extracorporeal life support. AB - The need for postcardiotomy mechanical support is uncommon and likely to decline. A mixture of options is necessary to meet the diverse indications for cardiac support in a comprehensive heart failure program. Between January 1997 and December 2000, 29 adult, neonate, and infant cardiac surgical patients were supported on an extracorporeal life support system. Indications for cardiac assist included post-cardiotomy low cardiac output syndrome, and hyperacute rejection after cardiac transplantation. Data for analysis were collected prospectively. Survival on the life support system was 20/29 (69%) and 12 patients (41%) survived to discharge. The mean time to starting extracorporeal life support was longer in survivors than non-survivors. The extracorporeal life support system provides effective cardiopulmonary and end-organ support. PMID- 17130325 TI - Surgical outcome of heterotaxy syndrome in a single institution. AB - Heterotaxy syndrome encompasses a wide range of anatomical variants including poor pulmonary vascular development. Surgical outcome remains poor in this difficult subgroup. Between April 1996 and November 2004, 27 patients with visceral heterotaxy were enrolled in this study. The median age at presentation was 25 days. There were 11 patients with asplenia and 16 with polysplenia. There were 5 deaths (4 in patients with asplenia, 1 in a patient with polysplenia). All patients with asplenia were scheduled for single-ventricle repair, and 5 completed a modified Fontan procedure. Of the 16 patients with polysplenia, 8 completed 2-ventricle repair and 6 had a modified Fontan. In polysplenia, actuarial survival was 93.8% at 4 months, and remained constant thereafter. In asplenia, survival was 81.8% at 1 month, and 53.0% at 1 year. A significant correlation was found between nonconfluent pulmonary arteries and mortality. The medium-term result in polysplenia was satisfactory, but not in asplenia. Nonconfluent pulmonary arteries carry a high risk of mortality, and a strategy to create pulmonary vascular confluence and satisfactory pulmonary blood flow is mandatory. PMID- 17130326 TI - Effect of pH management on brain perfusion during retrograde cerebral perfusion. AB - This study was undertaken to determine the effects of different pH management strategies during retrograde cerebral perfusion on the relationship between retrograde perfusion pressure and brain tissue perfusion. Six pigs were subjected to an alpha-stat strategy and another 6 to a pH-stat strategy during hypothermic (15 degrees C) retrograde cerebral perfusion at perfusion pressures of 10 to 70 mm Hg, in increments of 10 mm Hg every 20 min. Regional cerebral blood flow was significantly higher in the pH-stat group than in the alpha-stat group. The cerebral blood flow peaked at perfusion pressures of 40-50 mm Hg (18.6% +/- 10.8% in the pH-stat group vs. 3.6% +/- 1.2% in the alpha-stat group). In both groups, the intracranial pressure remained below the critical level of 25 mm Hg, even at a retrograde perfusion pressure of 70 mm Hg. Cerebral lactate production was higher in the alpha-stat group than the pH-stat group during retrograde cerebral perfusion at pressures of 10-30 mm Hg. Compared to the alpha-stat strategy, the pH-stat strategy significantly improved brain tissue perfusion. With an open inferior vena cava, the optimal perfusion pressure seems to be 40-50 mm Hg. PMID- 17130327 TI - Valved patch for ventricular septal defect with pulmonary arterial hypertension. AB - From March 1998 to December 2004, 16 acyanotic patients aged 2 to 22 years (mean, 7 +/- 5.7 years) with a large ventricular septal defect and elevated pulmonary vascular resistance (9.6 +/- 3.8 Wood units) underwent surgery. A Gore-Tex patch with a 5-8 mm longitudinal slit in the center was used. A piece of pericardium was sewn around the slit on one side of the patch, except for the upper quarter. In all patients, the defect was closed with a trimmed patch and the pericardial aspect was placed on the left ventricular side to allow right-to-left shunting. Echocardiography on the day of operation revealed a right-to-left shunt in 6 cases. Two patients (12.5%) died in the early postoperative period due to frequent episodes of pulmonary hypertensive crisis and persistent severe pulmonary hypertension. In 3 years of follow-up, pulmonary vascular resistance gradually decreased in all but one patient in whom it increased with a right-to left shunt and cyanosis. Insertion of a valved patch seems to be a promising technique to decrease morbidity and mortality in severe pulmonary arterial hypertension. PMID- 17130328 TI - Minor infection encouraged by steroid administration during cardiac surgery. AB - The aim of this study was to investigate whether steroid administration would increase the risk of postoperative infection. Sixty adults who underwent elective cardiac surgery under cardiopulmonary bypass were prospectively randomized into two groups. Thirty-one patients received hydrocortisone (50 mg x kg(-1)) before and after cardiopulmonary bypass, the other 29 served as controls. Various hemodynamic and pulmonary measurements were obtained perioperatively, and the white blood cell counts and levels of C-reactive protein were checked up to the 14(th) postoperative day. Steroid administration did not have any favorable effects during the perioperative period. Re-administration of antibiotics was needed in 7 patients (22.6%) after the 7(th) postoperative day in the steroid group, and in 3 (10.3%) in the control group. The peak white cell counts and C reactive protein levels after the 7(th) postoperative day were significantly higher in the steroid group. Steroid administration offered no clinical benefit to patients undergoing cardiac surgery with cardiopulmonary bypass, and it may encourage minor infections in the late postoperative period. PMID- 17130329 TI - Poorly differentiated primary monophasic synovial sarcoma of the lung. AB - Primary monophasic synovial sarcoma of the lung is rarely seen in clinical practice. We report the case of a 60-year-old male who underwent a left lower lobectomy for lung sarcoma. The patient received adjuvant therapy after surgery. Diagnosis was confirmed with SYT-SSX2 translocation detection. PMID- 17130330 TI - Tracheo-innominate artery fistula in children with high-lying innominate artery. AB - Tracheo-innominate artery fistula (TIF) is an uncommon but frequently fatal complication of tracheostomy. Significant airway hemorrhage usually occurs after premonitory bleeding. When massive bleeding occurs, immediate control of arterial bleeding, control of the airway and subsequent definite treatment are the principles for saving lives. Without prompt surgical intervention, the outcome of this complication is grave. Physicians should maintain a high index of suspicion of TIF in any patient with a recent tracheostomy and subsequent tracheal hemorrhage. PMID- 17130331 TI - Disseminated intravascular coagulopathy caused by intracardiac mesothelioma. AB - We report a case of intracardiac mesothelioma complicated by chronic disseminated intravascular coagulopathy in a 50-year-old woman. Her symptoms were completely relieved by emergency resection of the tumor. Primary resection of the intracardiac mesothelioma is adequate treatment for this complicated surgical problem. PMID- 17130332 TI - Subcutaneous metastasis from esophageal cancer. AB - Cutaneous metastases from esophageal adenocarcinoma are exceedingly rare. With the institution of multimodality treatments and surgical resection for esophageal adenocarcinoma, loco-regional disease recurrence has been dramatically reduced. In this report, we present a 59-year-old white male with adenocarcinoma of the distal esophagus that was treated with neoadjuvant chemotherapy and radiation therapy, followed by surgical excision. After a disease free interval of two years the patient presented with cutaneous metastatic disease. PMID- 17130333 TI - Cardiac transplantation and aortic coarctation repair in severe heart failure. AB - A 37-year-old man presented with severe dilated cardiomyopathy secondary to occult aortic coarctation. He was successfully managed with combined orthotopic heart transplantation and aortic coarctation repair. PMID- 17130334 TI - Late presentation of congenital Bochdalek hernia: a case report. AB - A 23-year-old man presented with a long history of dyspnea and wheezing thought to be due to asthma. Abnormal appearance of the left hemithorax was an incidental finding on a chest X-Ray. On further investigations he was found to have congenital Bochdalek hernia which was repaired surgically. All his respiratory symptoms resolved and he was able to discontinue treatment for asthma. We want to emphasise that late presentations can be misleading even to an astute clinician. PMID- 17130335 TI - Effective bleeding control during resection of giant carotid body tumor. AB - Carotid body tumor is a rare neoplasm located at the carotid bifurcation. Ligation and excision of the external carotid artery together with the tumor is preferred in patients with transmural tumor invasion. In those without transmural tumor invasion, temporary occlusion of the external carotid artery at the bifurcation allows trouble-free tumor excision and keeps the external carotid artery intact. PMID- 17130336 TI - Plasma troponin levels after cardiac surgery vs after myocardial infarction. AB - Raised plasma troponin, a diagnostic marker for myocardial infarction, usually occurs after cardiac surgery, leading to difficulties in diagnosing postoperative myocardial infarction. To ascertain whether the same processes influence troponin elevation in both conditions, a literature search was performed for plasma troponin elimination curves after myocardial infarction, myocardial infarction with reperfusion, and cardiac surgery. From 70 studies, 11 curves using the Stratus immunoassay kit were analyzed: 5 post-cardiac surgery (412 patients), 2 after myocardial infarction with reperfusion (169 patients), and 4 after myocardial infarction (640 patients). For each group, a new plot was formulated from the mean troponin level at each time interval. While the up-slope of the cardiac surgery curve was much steeper than that of myocardial infarction, resembling that of myocardial infarction with reperfusion, its down-slope was significantly more gentle than that of both other groups (-0.91 vs -5.31, t = 3.47, df = 8, p < 0.01). This suggests that postoperative troponin elevation involves enhanced cell permeability as seen after ischemia reperfusion rather than permanent cellular damage. The gentler down-slope may point to surgery induced impaired troponin removal from the circulation. Due to the different mechanisms proposed, implications from post-myocardial infarction troponin levels may not be conferred on post-cardiac surgery patients. PMID- 17130337 TI - Enucleation of giant leiomyoma of the esophagus. PMID- 17130338 TI - Combined off-pump coronary artery surgery and lung resection through thoracotomy. PMID- 17130339 TI - Left ventricle rupture after mitral valve replacement. PMID- 17130340 TI - Cardiovascular risk reduction in high-risk pediatric patients: a scientific statement from the American Heart Association Expert Panel on Population and Prevention Science; the Councils on Cardiovascular Disease in the Young, Epidemiology and Prevention, Nutrition, Physical Activity and Metabolism, High Blood Pressure Research, Cardiovascular Nursing, and the Kidney in Heart Disease; and the Interdisciplinary Working Group on Quality of Care and Outcomes Research: endorsed by the American Academy of Pediatrics. AB - Although for most children the process of atherosclerosis is subclinical, dramatically accelerated atherosclerosis occurs in some pediatric disease states, with clinical coronary events occurring in childhood and very early adult life. As with most scientific statements about children and the future risk for cardiovascular disease, there are no randomized trials documenting the effects of risk reduction on hard clinical outcomes. A growing body of literature, however, identifies the importance of premature cardiovascular disease in the course of certain pediatric diagnoses and addresses the response to risk factor reduction. For this scientific statement, a panel of experts reviewed what is known about very premature cardiovascular disease in 8 high-risk pediatric diagnoses and, from the science base, developed practical recommendations for management of cardiovascular risk. PMID- 17130341 TI - Alcohol consumption and risk of heart failure in the Physicians' Health Study I. AB - BACKGROUND: Heart failure (HF) is the leading cause of hospitalization among the elderly, and 1 in 5 adults aged 40 years will develop HF in their lifetime. Data on the effects of moderate alcohol consumption on the risk of HF have been sparse and inconsistent. This study sought to evaluate the association between moderate alcohol consumption and incident HF. METHODS AND RESULTS: A total of 21,601 participants of the Physicians' Health Study I who were free of HF and provided data on alcohol intake at baseline were prospectively followed up from 1982 to 2005. Incident HF cases were ascertained through annual follow-up questionnaires and validated with the use of Framingham criteria. During an average follow-up of 18.4 years, 904 incident cases of HF occurred. The crude incidence rates of HF were 25.0, 20.0, 24.3, and 20.6 cases per 10,000 person-years for alcohol categories of <1, 1 to 4, 5 to 7, and >7 drinks per week, respectively. Corresponding hazard ratios (95% CI) were 1.0 (reference), 0.90 (0.76 to 1.07), 0.84 (0.71 to 0.99), and 0.62 (0.41 to 0.96), respectively, with P for trend=0.012 adjusted for age, body mass index, smoking, and history of valvular heart disease. There was no evidence for a strong association between moderate alcohol consumption and HF without antecedent coronary artery disease. CONCLUSIONS: Although heavy drinking should be discouraged, our data indicate that moderate drinking may lower the risk of HF. The lack of an association between moderate alcohol intake and HF without antecedent coronary artery disease suggests that possible benefits of moderate drinking on HF may be mediated through beneficial effects of alcohol on coronary artery disease. PMID- 17130342 TI - Theoretical impact of the injection of material into the myocardium: a finite element model simulation. AB - BACKGROUND: To treat cardiac injuries created by myocardial infarcts, current approaches seek to add cells and/or synthetic extracellular matrices to the damaged ventricle to restore function. Because definitive myocardial regeneration remains undemonstrated, we propose that cardiac changes observed from implanted materials may result from altered mechanisms of the ventricle. METHODS AND RESULTS: We exploited a validated finite element model of an ovine left ventricle with an anteroapical infarct to examine the short-term effect of injecting material to the left ventricular wall. The model's mesh and regional material properties were modified to simulate expected changes. Three sets of simulations were run: (1) single injection to the anterior border zone; (2) therapeutic multiple border zone injections; and (3) injection of material to the infarct region. Results indicate that additions to the border zone decrease end-systolic fiber stress proportionally to the fractional volume added, with stiffer materials improving this attenuation. As a potential therapy, small changes in wall volume (approximately 4.5%) reduce elevated border zone fiber stresses from mean end-systole levels of 28.2 kPa (control) to 23.3 kPa (treatment), similar to levels of 22.5 kPA computed in remote regions. In the infarct, injection improves ejection fraction and the stroke volume/end-diastolic volume relationship but has no effect on the stroke volume/end-diastolic pressure relationship. CONCLUSIONS: Simulations indicate that the addition of noncontractile material to a damaged left ventricular wall has important effects on cardiac mechanics, with potentially beneficial reduction of elevated myofiber stresses, as well as confounding changes to clinical left ventricular metrics. PMID- 17130343 TI - High-dose allopurinol improves endothelial function by profoundly reducing vascular oxidative stress and not by lowering uric acid. AB - BACKGROUND: Allopurinol has been shown to improve endothelial function in chronic heart failure. This study aimed to establish its mechanism of action and to construct a dose-response curve for the effect of allopurinol. METHODS AND RESULTS: Two randomized, placebo-controlled, double-blind, crossover studies were performed for 1 month on patients with New York Heart Association Class II-III chronic heart failure, comparing 300 mg allopurinol, 600 mg allopurinol, and placebo for the first study and 1000 mg probenecid versus placebo in the second study. Endothelial function was assessed by standard forearm venous occlusion plethysmography. Allopurinol 600 mg/d significantly increased forearm blood flow response to acetylcholine compared with both allopurinol 300 mg/d and placebo (% change in forearm blood flow [mean+/-SEM]: 240.31+/-38.19% versus 152.10+/-18.21% versus 73.96+/-10.29%, P<0.001). For similar levels of urate lowering, the uricosuric agent probenecid had no effect on endothelial function. Sodium nitroprusside response was unchanged by all treatments. Vitamin C and acetylcholine coinfusion data showed that 600 mg/d allopurinol completely abolished the oxidative stress that was sensitive to high-dose vitamin C. CONCLUSIONS: For the first time, we have shown that a steep dose-response relationship exists between allopurinol and its effect on endothelial function. We also showed that the mechanism of improvement in endothelial function with allopurinol lies in its ability to reduce vascular oxidative stress and not in urate reduction. The reduction in vascular oxidative stress was profound because high-dose allopurinol totally abolished the oxidative stress that was sensitive to the high-dose vitamin C that was used in this study. PMID- 17130344 TI - Ethnic and gender differences in ambulatory blood pressure trajectories: results from a 15-year longitudinal study in youth and young adults. AB - BACKGROUND: Cross-sectional studies demonstrated ethnic and gender differences in ambulatory blood pressure patterns, but little is known about the longitudinal development of these differences. METHODS AND RESULTS: Twenty-four-hour ambulatory blood pressure was measured up to 12 times (5 times on average) over a 15-year period in 312 African Americans (AAs) and 351 European Americans aged 7 to 30 years. Multivariate individual growth curves across age were created for daytime and nighttime blood pressure jointly. For both daytime and nighttime systolic blood pressure (SBP), AAs and males had higher levels (P<0.001) than European Americans and females. Males also showed a greater increase with age (P<0.001) than females. For nighttime SBP, a faster increase of SBP with age (P<0.01) in AAs was additionally observed. The ethnic difference in nighttime SBP levels and its increase with age were significantly larger than in daytime SBP. For daytime and nighttime diastolic blood pressure, AAs had higher levels than European Americans (P<0.001), and this difference was significantly larger at night. From late adolescence onward, males showed a greater increase in diastolic blood pressure with age than females. Ethnic and gender differences persisted after adjustment for height, body mass index, socioeconomic status, and stress related coping styles. Family history of essential hypertension explained ethnic differences in daytime SBP. CONCLUSIONS: We observed significant ethnic and gender differences in longitudinal trajectories of ambulatory blood pressure in youth and young adults. The blunted nocturnal decline and its exacerbation with age in AAs corroborate and extend findings of cross-sectional studies. PMID- 17130345 TI - ACC/AHA/HRS 2006 key data elements and definitions for electrophysiological studies and procedures: a report of the American College of Cardiology/American Heart Association Task Force on Clinical Data Standards (ACC/AHA/HRS Writing Committee to Develop Data Standards on Electrophysiology). PMID- 17130346 TI - Stent-based local delivery of nuclear factor-kappaB decoy attenuates in-stent restenosis in hypercholesterolemic rabbits. AB - BACKGROUND: Nuclear factor-kappaB (NF-kappaB) plays a critical role in the vascular response to injury. However, the role of NF-kappaB in the mechanism of in-stent restenosis remains unclear. We therefore tested the hypothesis that blockade of NF-kappaB by stent-based delivery of a cis-element "decoy" of NF kappaB reduces in-stent neointimal formation. METHODS AND RESULTS: Stents were coated with a polymer containing or not containing NF-kappaB decoy, which represented a fast-release formulation (<7 days). Bare, polymer-coated, and NF kappaB decoy-eluting stents were implanted in iliac arteries of hypercholesterolemic rabbits. Increased NF-kappaB activity was noted at early stages after stenting, which was suppressed by stent-based delivery of NF-kappaB decoy. NF-kappaB decoy-eluting stents also reduced monocyte infiltration and monocyte chemoattractant protein-1 expression and suppressed CD14 activation on circulating leukocytes. Importantly, NF-kappaB decoy-eluting stents attenuated neointimal formation on day 28. There was no evidence of an incomplete healing process (persistent inflammation, hemorrhage, fibrin deposition, impaired endothelial regeneration) at the site of NF-kappaB decoy-eluting stents. Transfection of NF-kappaB decoy suppressed proliferation of human coronary artery smooth muscle cells in vitro. No systemic adverse effects of NF-kappaB decoy were detected. CONCLUSIONS: Stent-based local delivery of NF-kappaB decoy reduced in stent neointimal formation with no evidence of incomplete healing. These data suggest that this strategy may be a practical and promising means for prevention of in-stent restenosis in humans. PMID- 17130347 TI - Clopidogrel response variability and future therapies: clopidogrel: does one size fit all? PMID- 17130348 TI - Images in cardiovascular medicine. Giant atrial septal aneurysm in a 25-year-old woman. PMID- 17130349 TI - Images in cardiovascular medicine. Anomalous origin and course of the right coronary artery. PMID- 17130350 TI - Images in cardiovascular medicine. Intermittent loss of second heart sound. PMID- 17130351 TI - Letter by Jankowski regarding article, "Atherosclerotic lesion size and vulnerability are determined by patterns of fluid shear stress". PMID- 17130352 TI - Pioneers in cardiology: Salvador Moncada, MD, PhD, FRS. PMID- 17130353 TI - Marathoner's heart? PMID- 17130354 TI - Cardiac nociception. PMID- 17130355 TI - The scientific community's quest to identify optimal targets for anticoagulant pharmacotherapy. PMID- 17130356 TI - Detection and treatment of vulnerable plaques and vulnerable patients: novel approaches to prevention of coronary events. PMID- 17130357 TI - Left ventricular outflow obstruction: subaortic stenosis, bicuspid aortic valve, supravalvar aortic stenosis, and coarctation of the aorta. PMID- 17130359 TI - Sri Lanka: brand launch runs into protest. PMID- 17130360 TI - Corporate social responsibility: serious cause for concern. PMID- 17130362 TI - France: key role of NGOs in enforcement. PMID- 17130363 TI - Argentina: down Mexico way? PMID- 17130364 TI - Israel: first FCTC court case. PMID- 17130369 TI - Sidestream cigarette smoke toxicity increases with aging and exposure duration. AB - OBJECTIVES: To determine the effects of aging on the toxicity of sidestream tobacco smoke, the complex chemical mixture that enters the air from the lit end of burning cigarettes and constitutes the vast bulk of secondhand smoke. DESIGN: Statistical analysis of data from controlled experimental exposures of Sprague Dawley rats to fresh and aged (for more than 30 minutes) sidestream smoke for up to 90 days followed by histological sectioning of the respiratory epithelium. The data were obtained from a series of experiments conducted at Philip Morris' formerly secret INBIFO (Institut fur Biologische Forschung) laboratory in Germany. RESULTS: Using total particulate material as the measure of smoke exposure, aging sidestream cigarette smoke for at least 30 minutes increases its toxicity fourfold for 21 day exposures and doubles the toxicity for 90 day exposures, relative to fresh sidestream smoke. CONCLUSIONS: These results help explain the relatively large biological effects of secondhand smoke compared to equivalent mass doses of mainstream smoke. PMID- 17130370 TI - Marlboro UltraSmooth: a potentially reduced exposure cigarette? AB - AIM: To compare relative toxic emissions scores (RTE) of the carbon filter cigarette Marlboro UltraSmooth (MUS), against regular Marlboro, Holiday, and British Columbian brands. METHOD: MUS cigarettes were purchased in Tampa, Florida; Marlboro regular and Holiday were purchased in Auckland, New Zealand, and all emissions tested by Labstat International Inc, Kitchener, Ontario under Health Canada Intensive (HCI) machine-smoking conditions (55 ml puff per 30 seconds, filter ventilation holes blocked) against: (1) previous same brand emissions tested under ISO (International Organization for Standardization) conditions; (2) ISO and HCI average emissions for 16 regular brands sold in British Columbia (BC), the reference standard. Toxicants, selected by toxicological risk assessment, enabled estimation of an RTE per brand, and RTE per mg of nicotine. RESULTS: The BC standard for RTE in both ISO and HCI test modes, including metals and nitrosamines, was set at 100. Hereafter excluding them, RTE in ISO mode for BC was 97, MUS 4, Marlboro 102, and Holiday regular 99; and in HCI test mode BC was 97, MUS 42, Marlboro regular 107, and Holiday 95. From ISO to HCI, MUS total puff volume increased 50%, from 252 ml to 380 ml; nicotine yield increased 2.6 fold. Normalising for nicotine (RTE per mg nicotine), in ISO test mode, the BC standard was 97, MUS 10, Marlboro regular 124, and Holiday regular 107. In HCI mode, however, MUS/nicotine at 104 exceeded the average BC standard of 97; Marlboro regular was 137, and Holiday regular 97; MUS ranked sixth highest among 18 regular brands. MUS contained 103 mg of carbon in its 304 mg filter, which was 55% ventilated. CONCLUSION: The combined acetate carbon filter of MUS performed best at low smoke volumes on ISO testing. Under more smoker-realistic intensive machine testing, and correcting for relative nicotine concentration and compensatory smoking, MUS increased the RTE, for all toxicants combined, for carcinogens, and for cardiovascular toxicants, compared with most regular brands. MUS was not a potentially reduced-exposure product (PREP) under smoker-realistic test conditions, and thus would not be expected to reduce overall harm. It is unrealistic to expect that even major design changes, as seen in MUS, or a regulatory framework to enforce such changes, could reduce cigarette smoking mortality risks to acceptable levels. PMID- 17130371 TI - The temporal relationship between advertising and sales of low-tar cigarettes. AB - OBJECTIVE AND HYPOTHESIS: To determine whether a temporal relationship exists between the advertising and sales of low-tar cigarettes. It was hypothesised that increases in the advertising of low-tar cigarettes would precede increases in sales for these cigarettes. METHODS: The themes of cigarette advertisements were reviewed and coded for 20 low-tar cigarette brands advertised in 13 widely read magazines in the US between 1960 and 1996. These 20 brands represented most of the low-tar cigarette advertisements and cigarette sales from 1967 to 1996. Cigarette sales data were obtained from the 1994 Maxwell report that summarises all cigarette sales from 1925 to 1990. If the advertisement referred to the low tar attributes of the cigarette advertised, the advertisement was coded as having a low-tar theme and was included in the analysis. RESULTS: Five different graphical presentations of the relationship between the advertising and sales of the 20 low-tar cigarette brands showed a temporal relationship between low-tar advertising and sales for these brands. This relationship was observed for brands that introduced a low-tar alternative into an existing brand family (eg, Marlboro Light) and for new exclusively low-tar brands (eg, Carlton). Despite large increases in the advertising for the exclusively low-tar brands, sales of these brands remained low relative to sales of the low-tar alternative brands. CONCLUSIONS: Increases in print advertising of 20 of the most popular low-tar cigarette brands were followed by increases in sales for these cigarettes. Despite increases in the advertising of exclusively low-tar brands in the mid 1970s and early 1980s, the sales of these brands never matched the sales of the low-tar alternative brands. This suggests that it may have been easier to get smokers to switch to low-tar brands within a brand family compared with entirely new low-tar brands. Over the past 30 years, the marketing of low-tar cigarettes as a healthier alternative to higher-tar cigarettes has resulted in these brands dominating the market, and may have kept concerned smokers from quitting. PMID- 17130372 TI - Prevalence of smoking among major movie characters: 1996-2004. AB - BACKGROUND: Reports of a relationship between watching smoking in movies and smoking among adolescents have prompted greater scrutiny of smoking in movies by the public health community. OBJECTIVE: To assess the smoking prevalence among adult and adolescent movie characters, examine trends in smoking in movies over time, and compare the data with actual smoking prevalence among US adults and adolescents. DESIGN AND METHODS: Smoking status of all major human adolescent and adult movie characters in the top 100 box office hits from 1996 to 2004 (900 movies) was assessed, and smoking prevalence was examined by Motion Picture Association of America (MPAA) rating and year of release. RESULTS: The movies contained 5944 major characters, of whom 4911 were adults and 466 were adolescents. Among adult movie characters, the overall smoking prevalence was 20.6%; smoking was more common in men than in women (22.6% v 16.1%, respectively, p<0.001), and was related to MPAA rating category (26.9% for movies rated R (restricted, people aged <17 years require accompanying adult), 17.9% for PG-13 (parents strongly cautioned that some material might be inappropriate for children) and 10.4% for G/PG (general audiences, all ages; parental guidance suggested for children), p<0.001). In 1996, the smoking prevalence for major adult movie characters (25.7%) was similar to that in the actual US population (24.7%). Smoking prevalence among adult movie characters declined to 18.4% in 2004 (p for trend <0.001), slightly below that for the US population for that year (20.9%). Examination of trends by MPAA rating showed that the downward trend in smoking among adult movie characters was statistically significant in movies rated G/PG and R, but not in those rated PG-13. A downward trend over time was also found for smoking among adolescent movie characters. There was no smoking among adult characters in 43.3% of the movies; however, in 39% of the movies, smoking prevalence among adult characters was higher than that in the US adult population in the year of release. CONCLUSIONS: Smoking prevalence among major adolescent and adult movie characters is declining, with the downward trend among adult characters weakest for PG-13-rated movies. Although many movies depict no adult smoking, more than one third depict smoking as more prevalent than that among US adults at the time of release. PMID- 17130373 TI - The US Public Health Service "treating tobacco use and dependence clinical practice guidelines" as a legal standard of care. AB - BACKGROUND: The important factors in evaluating the role of clinical practice guidelines (CPGs) in medical malpractice litigation have been discussed for several years, but have focused on broad policy implications rather than on a concrete example of how an actual guideline might be evaluated. There are four items that need to be considered in negligence torts: legal duty, a breach of that duty, causal relationship between breach and injury, and damages. OBJECTIVE: To identify the arguments related to legal duty. RESULTS: The Treating Tobacco Use and Dependence (revised 2000) CPG, sponsored by the US Public Health Service, recommends effective and inexpensive treatments for nicotine addiction, the largest preventable cause of death in the US, and can be used as an example to focus on important considerations about the appropriateness of CPGs in the judicial system. Furthermore, the failure of many doctors and hospitals to deal with tobacco use and dependence raises the question of whether this failure could be considered malpractice, given the Public Health Service guideline's straightforward recommendations, their efficacy in preventing serious disease and cost-effectiveness. CONCLUSION: Although each case of medical malpractice depends on a multitude of factors unique to individual cases, a court could have sufficient basis to find that the failure to adequately treat the main cause of preventable disease and death in the US qualifies as a violation of the legal duty that doctors and hospitals owe to patients habituated to tobacco use and dependence. PMID- 17130374 TI - Does maternal smoking during pregnancy predict the smoking patterns of young adult offspring? A birth cohort study. AB - OBJECTIVE: To examine the association between maternal smoking during pregnancy and the development of smoking behaviour patterns among young adult offspring. METHOD: Data were from the Mater-University of Queensland Study of Pregnancy (MUSP), a birth cohort of 7223 mothers and children enrolled in Brisbane, Australia, in 1981. The development of smoking behaviours (early or late onset, or combination of onset and prevalence patterns) among offspring at age 21 years with different patterns of maternal smoking (never smoked, smoked before or after pregnancy but not during pregnancy, or smoked during pregnancy) were compared. Maternal smoking information was derived from the prospectively collected data from the beginning of pregnancy until the child was 14 years of age. Analyses were restricted to the 3058 mothers and children whose smoking status was reported. RESULTS: The proportion of young adults who smoked regularly, either with early onset or late onset, was greater among those whose mothers had smoked during pregnancy compared with those whose mothers had never smoked. The smoking patterns among those adolescent offspring whose mothers stopped smoking during pregnancy, but who then smoked at other times during the child's life, were similar to those whose mothers had never smoked. This association was robust to adjustment for a variety of potential covariates. CONCLUSIONS: The findings provide some evidence for a direct effect of maternal smoking in utero on the development of smoking behaviour patterns of offspring and provide yet another incentive to persuade pregnant women not to smoke. PMID- 17130375 TI - The association of retail promotions for cigarettes with the Master Settlement Agreement, tobacco control programmes and cigarette excise taxes. AB - BACKGROUND: Retail stores are the primary medium for marketing cigarettes to smokers in the US. The prevalence and characteristics of cigarette retail advertising and promotions have been described by several investigators. Less is known about the proportion of cigarette sales occurring as part of a retail promotion and about the effects of tobacco control policies on cigarette promotions. OBJECTIVE: To estimate the effect of the Master Settlement Agreement (MSA), state tobacco control programme funding and cigarette taxes on retail promotions for cigarettes in supermarkets in the US. OUTCOME MEASURES: Proportion of cigarette sales occurring under a retail promotion and the value of multipack promotions (eg, buy one pack, get one pack free) and cents-off promotions, measured using scanner data in supermarkets from 50 retail market areas from 1994 to 2004. RESULTS: Promoted cigarette sales have increased significantly since the MSA (p<0.01), and are higher in market areas with high tobacco control programme funding (p<0.01) and high cigarette tax (p<0.01). The value of a multipack promotion is higher since the MSA (p<0.01) and in market areas with high cigarette tax (p<0.01). The value of a cents-off promotion is negatively related to the MSA (p<0.01), with mixed results for tobacco control programme funding (p<0.05), and is unassociated with tax. CONCLUSIONS: Higher promoted cigarette sales and increased promotional values in market areas with strong tobacco control policies, compared with market areas with weaker tobacco control policies, may partially offset the decline in smoking achieved in those areas. PMID- 17130376 TI - Mortality, morbidity and costs attributable to smoking in Germany: update and a 10-year comparison. AB - OBJECTIVE: To assess the negative health consequences and associated costs of cigarette smoking in Germany in 2003 and to compare them with the respective results from 1993. METHODS: The number of deaths, years of potential life lost (YPLL), direct medical and indirect costs caused by active cigarette smoking in Germany in 2003 is estimated from a societal perspective. The method is similar to that applied by Welte et al, who estimated the cost of smoking in Germany in 1993. Therefore, a direct comparison of the results was possible. Methodological and data differences between these two publications and their effect on the results are analysed. RESULTS: In 2003, 114,647 deaths and 1.6 million YPLL were attributable to smoking. Total costs were euro21.0 billion, with euro7.5 billion for acute hospital care, inpatient rehabilitation care, ambulatory care and prescribed drugs; euro4.7 billion for the indirect costs of mortality; and euro8.8 billion for costs due to work loss days and early retirement. From 1993 to 2003, the proportionate mortality attributable to smoking remained relatively stable, rising from 13.0% to 13.4%. The smoking-attributable deaths in men is lowered by 13.7% whereas that in women increased by 45.3%. Total real direct costs rose by 35.8%, and total real indirect costs declined by 7.1%, rendering an increase of 4.7% to real total costs. Accountable factors are changes in cigarette smoking prevalence and in disease-specific mortality and morbidity, as well as a rise in general healthcare expenditure. CONCLUSIONS: Despite the growing knowledge about the hazards of smoking, the smoking-attributable costs increased in Germany. Further, female mortality attributable to smoking is much higher than it was in 1993. PMID- 17130377 TI - Health consequences of reduced daily cigarette consumption. AB - OBJECTIVE: To determine the risk of dying from specified smoking-related diseases and from any cause in heavy smoking men and women (> or =15 cigarettes/day), who reduced their daily cigarette consumption by >50%. DESIGN: A prospective cohort study. SETTING: Three counties in Norway. PARTICIPANTS: 24,959 men and 26,251 women, aged 20-49 years, screened for risk factors of cardiovascular disease in the mid-1970s, screened again after 3-13 years, and followed up throughout 2003. OUTCOMES: Absolute mortality and relative risks adjusted for confounding variables, of dying from all causes, cardiovascular disease, ischaemic heart disease, all smoking-related cancer and lung cancer. RESULTS: With sustained heavy smokers as reference, the smokers of both sexes who reduced their daily consumption (reducers) had the following adjusted relative risks (95% confidence interval (CI)): of dying from any cause, 1.02 (0.84 to 1.22); cardiovascular disease, 1.02 (0.75 to 1.39); ischaemic heart disease, 0.96 (0.65 to 1.41); smoking-related cancer, 0.86 (0.57 to 1.29); and lung cancer, 0.66 (0.36 to 1.21). The difference in cigarette consumption between two examinations was not a significant predictor of death from any of the causes. A follow-up from a third screening of the subgroup who were reducers at both second and third examinations (sustained reducers) did not have a lower risk than those who were heavy smokers at all three examinations. CONCLUSIONS: Long-term follow-up provides no evidence that heavy smokers who cut down their daily cigarette consumption by >50% reduce their risk of premature death significantly. In health education and patient counselling, it may give people false expectations to advise that reduction in consumption is associated with reduction in harm. PMID- 17130378 TI - An uncontrolled trial of cytisine (Tabex) for smoking cessation. AB - OBJECTIVES: Cytisine (Tabex) has been licensed in Eastern Europe as an aid to smoking cessation for 40 years. Cytisine is a partial agonist with high affinity binding to the alpha4beta2 nicotinic acetylcholine receptor believed to be central to the rewarding effect of nicotine. There is insufficient information on effectiveness to warrant licensing by modern standards. To assess whether full scale controlled trials are warranted, this study sought to obtain an estimate of the 12-month continuous abstinence rates of smokers using cytisine with minimal behavioural support. DESIGN: An uncontrolled, open-label trial. SETTING: A smokers' clinic in an oncology centre in Warsaw, Poland. SUBJECTS: 436 consecutive attendees of the smokers' clinic of whom 191 were male. The mean dependence score (Fagerstrom Test for Nicotine Dependence) was 6.1. INTERVENTION: The standard regimen of Tabex (cytisine) was used, involving 25 days of treatment with minimal behavioural support. MAIN OUTCOME MEASURE: Self-reported continuous abstinence for 12 months; with abstinence verified by carbon monoxide at the final follow up (after 12 months). RESULTS: 60 participants (13.8% of the total sample) were abstinent for 12 months. Of the 315 subjects, who had taken the drug, 49 (15.5%) stopped cytisine because of adverse effects (mostly gastric disturbances and nausea), although they were not serious. The frequency of the minor adverse effects, primarily gastric disturbance, was similar to that observed in previous studies with the drug. CONCLUSIONS: The long-term abstinence rates were similar to those observed in smokers receiving nicotine replacement therapy. Full-scale randomised trials of cytisine (Tabex), conducted to the standards required by regulatory authorities, are warranted. PMID- 17130379 TI - Smokers living in deprived areas are less likely to quit: a longitudinal follow up. AB - OBJECTIVE: To follow up smokers to examine whether the likelihood of quitting smoking varied by area deprivation, and whether smoking history, health status, personality characteristics, social support and stressful situations contributed to differences in area deprivation in quit rates. DESIGN: Longitudinal data with a 6-year follow-up period were analysed using multilevel logistic regression. Area-level deprivation was characterised by a composite measure that was the sum of the proportion of unemployed residents, the percentage of residents in blue collar occupations and the proportion with only elementary-level education. Previously established predictors of smoking cessation, including education, age at smoking initiation, self-assessed health, chronic illness, locus of control, neuroticism, negative life events, longlasting relationship difficulties, emotional social support and negative neighbourhood conditions were examined separately and in a combined model to assess whether they contributed to neighbourhood deprivation differences in quitting. PARTICIPANTS: 404 participants (residing in 83 areas) identified as smokers at baseline and who did not change their residential address over the follow-up period. MAIN OUTCOME: Being a non smoker at follow-up. RESULTS: Odds ratios of quitting decreased with greater area level deprivation, but differences reached significance only between the most and least deprived quartiles. Smoking history, health status, personality characteristics, social support and stressful situations did not contribute to the lower quitting rates seen among smokers in deprived areas. CONCLUSIONS: Living in a deprived area seems to reduce the likelihood of quitting smoking; hence individual-level tobacco control efforts should be complemented with area based interventions. However, we need to identify and understand the underlying factors associated with living in a deprived area that contributes to lower quitting rates. PMID- 17130381 TI - Collaborative care models for depression: time to move from evidence to practice. PMID- 17130382 TI - Primary prevention of cardiovascular diseases with statin therapy: a meta analysis of randomized controlled trials. AB - BACKGROUND: While the role of hydroxymethyl glutaryl coenzyme A reductase inhibitors (statins) in secondary prevention of cardiovascular (CV) events and mortality is established, their value for primary prevention is less clear. To clarify the role of statins for patients without CV disease, we performed a meta analysis of randomized controlled trials (RCTs). METHODS: MEDLINE, EMBASE, Cochrane Collaboration, and American College of Physicians Journal Club databases were searched for RCTs published between 1966 and June 2005. We included RCTs with follow-up of 1 year or longer, more than 100 major CV events, and 80% or more of the population without CV disease. From each trial, demographic data, lipid profile, CV outcomes, mortality, and adverse outcomes were recorded. Summary relative risk (RR) ratios with 95% confidence intervals (CIs) were calculated using a random effects model. RESULTS: Seven trials with 42,848 patients were included. Ninety percent had no history of CV disease. Mean follow up was 4.3 years. Statin therapy reduced the RR of major coronary events, major cerebrovascular events, and revascularizations by 29.2% (95% CI, 16.7%-39.8%) (P<.001), 14.4% (95% CI, 2.8%-24.6%) (P = .02), and 33.8% (95% CI, 19.6%-45.5%) (P<.001), respectively. Statins produced a nonsignificant 22.6% RR reduction in coronary heart disease mortality (95% CI, 0.56-1.08) (P = .13). No significant reduction in overall mortality (RR, 0.92 [95% CI, 0.84-1.01]) (P = .09) or increases in cancer or levels of liver enzymes or creatine kinase were observed. CONCLUSION: In patients without CV disease, statin therapy decreases the incidence of major coronary and cerebrovascular events and revascularizations but not coronary heart disease or overall mortality. PMID- 17130383 TI - Collaborative care for depression: a cumulative meta-analysis and review of longer-term outcomes. AB - BACKGROUND: Depression is common in primary care but is suboptimally managed. Collaborative care, that is, structured care involving a greater role of nonmedical specialists to augment primary care, has emerged as a potentially effective candidate intervention to improve quality of primary care and patient outcomes. METHODS: To quantify the short-term and longer-term effectiveness of collaborative care compared with standard care and to understand mechanisms of action by exploring between-study heterogeneity, we conducted a systematic review of randomized controlled trials that compared collaborative care with usual primary care in patients with depression. We searched MEDLINE (from the beginning of 1966), EMBASE (from the beginning of 1980), CINAHL (from the beginning of 1980), PsycINFO (from the beginning of 1980), the Cochrane Library (from the beginning of 1966), and DARE (Database of Abstracts of Reviews of Effectiveness) (from the beginning of 1985) databases from study inception to February 6, 2006. RESULTS: We found 37 randomized studies including 12 355 patients with depression receiving primary care. Random effects meta-analysis showed that depression outcomes were improved at 6 months (standardized mean difference, 0.25; 95% confidence interval, 0.18-0.32), and evidence of longer-term benefit was found for up to 5 years (standardized mean difference, 0.15; 95% confidence interval, 0.001-0.31). When exploring determinants of effectiveness, effect size was directly related to medication compliance and to the professional background and method of supervision of case managers. The addition of brief psychotherapy did not substantially improve outcome, nor did increased numbers of sessions. Cumulative meta-analysis showed that sufficient evidence had emerged by 2000 to demonstrate the statistically significant benefit of collaborative care. CONCLUSIONS: Collaborative care is more effective than standard care in improving depression outcomes in the short and longer terms. Future research needs to address the implementation of collaborative care, particularly in settings other than the United States. PMID- 17130384 TI - Risk of new-onset atrial fibrillation in relation to body mass index. AB - BACKGROUND: Obesity is associated with increased risk of atrial fibrillation (AF), but it is unknown whether the association differs by duration or persistence of AF. It is also unknown to what extent cardiovascular risk factors may mediate this association. METHODS: This population-based case-control study included 425 subjects with new-onset AF and 707 controls. The AF cases were identified through International Classification of Diseases, Ninth Revision codes for inpatient and outpatient visits and verified by medical record review. Medical records provided data on height, weight, and cardiovascular risk factors. RESULTS: On average, AF risk was 3% higher (95% confidence interval [CI], 1%-5%) per unit increment in body mass index (BMI) (calculated as weight in kilograms divided by height in meters squared). For sustained AF (duration > or =6 months), risk was higher by 7% (95% CI, 3%-11%) per unit BMI increment; for intermittent AF (duration > or =8 days or recurrent), 4% (95% CI, 1%-6%); and for transitory AF (duration <8 days), 1% (95% CI, -1% to +4%). Compared with those with normal BMI, the odds ratios for overweight and obese subjects were as follows: overweight, 0.97 (95% CI, 0.68-1.38); obese class 1, 1.18 (95% CI, 0.80-1.73); obese class 2, 1.34 (95% CI, 0.82-2.18); and obese class 3, 2.31 (95% CI, 1.36 3.91) (P = .002 for trend). When diabetes mellitus, a possible mediator, was added to the model, the odds ratio per unit increment of BMI decreased from 1.034 to 1.028. Adjustment for other cardiovascular risk factors including hyperlipidemia and blood pressure did not attenuate the BMI-AF association. CONCLUSIONS: The association with BMI was stronger for sustained AF than for transitory or intermittent AF. The obesity-AF association appears to be partially mediated by diabetes mellitus but minimally through other cardiovascular risk factors. PMID- 17130385 TI - Safety of exercise training for cardiac patients: results of the French registry of complications during cardiac rehabilitation. AB - BACKGROUND: Cardiac rehabilitation is widely recognized as a medical management procedure that reduces mortality, but the cardiovascular safety of exercise training has not been clearly established. Published data are retrospective or outdated, as patient management has substantially progressed in recent years. The aim of this prospective registry was to determine the current complication rate during exercise performed in the course of cardiac rehabilitation. METHODS: This study was conducted by the Functional Evaluation and Cardiac Rehabilitation Working Group of the French Society of Cardiology. During a 1-year period, 65 cardiac rehabilitation centers reported that serious events had occurred during or 1 hour after an exercise stress test or a training session. Severe cardiovascular events were validated by a scientific committee. RESULTS: A total of 25,420 patients (78% men; mean age, 61.3 years) were included in the study. Initial indications for cardiac rehabilitation were post-cardiac surgery (coronary bypass, 34.3%; valvular surgery, 18.4%); recent percutaneous coronary intervention (21.6%); and other coronary (13.2%) and noncoronary (12.5%) conditions. The study population underwent 42,419 exercise stress tests and 743,471 patient-hours of exercise training. Twenty severe cardiac events were reported: 5 were related to exercise testing and 15 were related to exercise training. The event rate was 1 per 8484 exercise stress tests and 1 per 49,565 patient-hours of exercise training; the cardiac arrest rate was 1.3 per million patient-hours of exercise. Neither fatal complications nor emergency defibrillations were reported. CONCLUSION: The frequency of major cardiovascular complications during supervised exercise training in France is quite low. PMID- 17130386 TI - Low micronutrient levels as a predictor of incident disability in older women. AB - BACKGROUND: The role of nutritional status in the disablement process is still unclear. The objective of this study was to assess whether low concentrations of nutrients predict the development and course of disability. METHODS: Longitudinal study including community-dwelling women 65 years or older enrolled in the Women's Health and Aging Study I. In total, 643 women were assessed prospectively at 6-month intervals from 1992 to 1995. RESULTS: Incidence rates of disability in activities of daily living (ADLs) during 3 years of follow-up. Incidence rates in the lowest quartile of each selected nutrient were compared with those in the upper quartiles. The hazard ratios were estimated from Cox models adjusted for potential confounders. Women in the lowest quartile of serum concentrations of vitamin B(6) (hazard ratio [HR], 1.31; 95% confidence interval [CI], 1.03-1.67), vitamin B(12) (HR, 1.40; 95% CI, 1.12-1.74), and selenium (HR, 1.38; 95% CI, 1.12 1.71) had significantly higher risk of disability in ADLs during 3 years of follow-up compared with women in the upper 3 quartiles. CONCLUSIONS: Low serum concentrations of vitamins B(6) and B(12) and selenium predict subsequent disability in ADLs in older women living in the community. Nutritional status is one of the key factors to be considered in the development of strategies aimed at preventing or delaying the disablement process. PMID- 17130387 TI - Prediction of coronary artery calcium in young adults using the Pathobiological Determinants of Atherosclerosis in Youth (PDAY) risk score: the CARDIA study. AB - BACKGROUND: Using data from autopsied young people aged 15 to 34 years, the Pathobiological Determinants of Atherosclerosis in Youth (PDAY) study developed a risk score based on age, sex, smoking status, high-density lipoprotein and non high-density lipoprotein cholesterol levels, and the presence of obesity, hyperglycemia, and hypertension to predict advanced coronary artery atherosclerosis. METHODS: The Coronary Artery Risk Development in Young Adults (CARDIA) study assessed coronary artery calcium (CAC) by computed tomography in young adults participating in the 15-year examination. The PDAY risk score was calculated from risk factors measured at the CARDIA examinations at years 0, 5, 10, and 15. RESULTS: Odds ratios for amount of CAC (6 ordinal categories) for a 1 point increase in risk score computed from the modifiable risk factors ranged from 1.10 to 1.16 (all statistically significant). Odds ratios for presence of any amount of CAC ranged from 1.09 to 1.15 (all statistically significant), with the highest odds ratio for the risk score at year 0. An increase in risk score between years 0 and 15 increased the odds of CAC, and a decrease in risk score decreased the odds of CAC. A positive family history of cardiovascular disease increased the odds of CAC. The c statistics ranged from 0.752 to 0.770, with the highest discrimination based on the year 0 revised PDAY risk score that included family history and increased the points for the sex differential. CONCLUSION: The PDAY risk score predicts CAC up to 15 years before its assessment, and risk score change during 15 years affects the risk of CAC. PMID- 17130388 TI - Socioeconomic status and trends in disparities in 4 major risk factors for cardiovascular disease among US adults, 1971-2002. AB - BACKGROUND: It is unknown whether the previously recognized disparities in cardiovascular disease (CVD) risk factors related to annual income and educational level have diminished, persisted, or worsened in recent decades. The objective of this study was to examine 31-year trends in CVD risk factors by annual income and educational levels among US adults. METHODS: Four cross sectional national surveys were used: National Health and Nutrition Examination Survey I (1971-1974), II (1976-1980), III (1988-1994), and 1999-2002. The main outcome measure was prevalence of high cholesterol (> or =240 mg/dL [> or =6.2 mmol/L]), high blood pressure (140/90 mm Hg), smoking, and diabetes mellitus. RESULTS: Between 1971 and 2002, the prevalence of all CVD risk factors, except diabetes, decreased in all income and education groups, but there has been little reduction in income- and education-related disparities in CVD risk factors and few improvements during the past 10 years. The prevalence of high blood pressure declined by about half in all income and education groups, ranging from 30.3% to 40.6% in 1971-1974 and 16.4% in 1999-2002, with the greatest reduction among those in the lowest income quartile and those with less than a high school education (18.0 and 15.9 percentage points, respectively). High cholesterol prevalence also declined in all groups and ranged from 28.8% to 32.4% in 1971 1974 and 15.3% to 22.0% in 1999-2002, with the largest decline (15.9 percentage points) among people with the highest incomes. Education- and income-related disparities in smoking widened considerably, because there were large declines in smoking prevalence among people with high incomes and education (from about 33% in 1971-1974 to about 14%-17% in 1999-2002) but only marginal reductions among those with low incomes and education (about 6-percentage point decline). Diabetes prevalence increased most among persons with low incomes and education. CONCLUSIONS: Despite the general success in reducing CVD risk factors in the US population, not all segments of society are benefiting equally and improvements may have slowed. Education- and income-related disparities have worsened for smoking, and increases in diabetes prevalence have occurred primarily among persons with a lower socioeconomic status. Diabetes prevention and smoking prevention and cessation programs need to specifically target persons of lower income and education. PMID- 17130389 TI - Association of childhood socioeconomic status with subsequent coronary heart disease in physicians. AB - BACKGROUND: Adult socioeconomic status (SES) is an independent risk factor for the development of coronary heart disease (CHD), but whether low childhood SES has an effect in adults who have achieved high SES is unknown. METHODS: We examined the risk of CHD and mortality associated with low childhood SES in 1131 male medical students from The Johns Hopkins Precursors Study, a prospective cohort of graduates of The Johns Hopkins University School of Medicine from 1948 to 1964 with a median follow-up of 40 years. RESULTS: Of 1131 subjects, 216 (19.1%) were from low-SES families. Medical students from low-SES families were slightly older at graduation (26.8 vs 26.2 years; P = .004) and gained more weight over time (P = .01). Low childhood SES conferred a 2.40-fold increased hazard of developing CHD on or before age 50 years (95% confidence interval, 1.21 4.74) but not at older ages. The impact of low SES on early CHD was not reduced by adjusting for other CHD risk factors, including body mass index, cholesterol level, amount of exercise, depression, coffee drinking, smoking, hypertension, diabetes mellitus, and parental CHD history. Low childhood SES did not confer an increased risk of all-cause mortality. CONCLUSIONS: Low childhood SES is associated with an increased incidence of CHD before age 50 years among men with high adulthood SES. This risk is not mediated by traditional risk factors for CHD. These findings highlight the importance of childhood events on the development of CHD early in adulthood and the persistent effects of low SES. PMID- 17130390 TI - Ischemic mitral regurgitation and risk of heart failure after myocardial infarction. AB - BACKGROUND: The development of ischemic mitral regurgitation (MR) after myocardial infarction may impose hemodynamic load during a period of active left ventricular remodeling and promote heart failure (HF). However, few data are available on the relationship between ischemic MR and the long-term risk for HF. METHODS: We prospectively studied 1190 patients admitted for acute myocardial infarction. Mitral regurgitation was assessed by echocardiography and was considered mild, moderate, and severe when the regurgitant jet area occupied less than 20%, 20% to 40%, and greater than 40% of the left atrial area, respectively. The median duration of follow-up was 24 months (range, 6-48 months). RESULTS: Mild and moderate or severe ischemic MR was present in 39.7% and 6.3% of patients, respectively. After adjusting for ejection fraction and clinical variables (age, sex, Killip class, previous infarction, hypertension, diabetes mellitus, anterior infarction, ST-elevation infarction, and coronary revascularization), compared with patients without MR, the hazard ratios for HF were 2.8 (95% confidence interval [CI], 1.8-4.2; P<.001) and 3.6 (95% CI, 2.0 6.4; P<.001) in patients with mild and moderate or severe ischemic MR, respectively. The adjusted hazard ratios for death were 1.2 (95% CI, 0.8-1.8; P = .43) and 2.0 (95% CI, 1.2-3.4; P = .02) in patients with mild and moderate or severe MR, respectively. CONCLUSIONS: There is a graded independent association between the severity of ischemic MR and the development of HF after myocardial infarction. Even mild ischemic MR is associated with an increase in the risk of HF. PMID- 17130391 TI - Weight cycling and risk of gallstone disease in men. AB - BACKGROUND: The long-term effect of repeated intentional weight loss and weight regain on the risk of gallstone disease in men is not clear. METHODS: Participants in the Health Professionals Follow-up Study provided information on intentional weight loss during the previous 4 years in 1992. Weight cyclers were men who had intentional weight loss and weight regain. Men free of gallstone disease at baseline were followed from 1992 to 2002. On biennial questionnaires the participants reported newly diagnosed gallstone disease. RESULTS: During 264,760 person-years of follow-up we ascertained 1222 cases of symptomatic gallstones. We examined the effect of weight cycling on the risk of gallstone disease. The multivariate relative risk of weight cyclers, compared with weight maintainers, after adjusting for potential confounding variables, including body mass index, was 1.11 (95% confidence interval [CI], 0.94-1.31) in light cyclers, 1.18 (95% CI, 0.97-1.43) in moderate cyclers, and 1.42 (95% CI, 1.11-1.81) in severe cyclers. We further examined the effect of number of cycling episodes. Among weight cyclers, the relative risk associated with having more than 1 weight cycle, compared with weight maintainers, was 1.10 (95% CI, 0.88-1.37) in light cyclers, 1.28 (95% CI, 1.03-1.59) in moderate cyclers, and 1.51 (95% CI, 1.13 2.02) in severe cyclers. CONCLUSIONS: Our findings suggest that weight cycling, independent of body mass index, may increase the risk of gallstone disease in men. Larger weight fluctuation and more weight cycles are associated with greater risk. PMID- 17130392 TI - Quality of cardiopulmonary resuscitation among highly trained staff in an emergency department setting. AB - BACKGROUND: Recent reports have highlighted the poor standard of cardiopulmonary resuscitation (CPR) achieved by health care professionals in diverse situations. We explored what can be achieved in an emergency department by highly trained permanent staff. METHODS: In a prospective observational study conducted from June 1, 2002, to August 31, 2005, 80 of 213 patients requiring CPR and admitted to the emergency department of a tertiary care hospital were eligible for study participation. Owing to several logistic problems with CPR, 133 patients could not be studied. The CPR team consisted of emergency- and critical care-trained physicians with more than 10 years of acute care experience, most of whom were instructors of European Resuscitation Council courses in basic and advanced life support. A specially designed defibrillator was used to assess the quality of CPR. RESULTS: For 80 patients, 95 data sets were available for analysis, yielding a total of 1065 minutes of cardiac arrest time. Chest compressions were performed at a rate of 114 (95% confidence interval [CI], 112-116) per minute, resulting in a mean of 96 (95% CI, 93-99) delivered chest compressions per minute. We further observed a mean hands-off ratio of 12.7% (95% CI, 12.3%-13.1%), and the hands-off ratio was linearly associated with the duration of CPR (R(2) = 0.95; mean, 4.3% increments per 5-10 minutes; P<.001). Patients were hyperventilated with a median of 18 (interquartile range, 14-24) ventilations per minute. CONCLUSIONS: Highly trained professionals in an emergency department can achieve appropriate chest compression rates during CPR with a low hands-off ratio. Increased attention must be paid in all situations to the avoidance of hyperventilation. PMID- 17130393 TI - Prevalence of lower urinary tract symptoms and effect on quality of life in a racially and ethnically diverse random sample: the Boston Area Community Health (BACH) Survey. AB - BACKGROUND: Previous studies of lower urinary tract symptoms (LUTS) have focused on men, with few studies including minority populations. The Boston Area Community Health (BACH) Survey is designed to study the prevalence and impact of LUTS among both men and women in a racially, ethnically, and socioeconomically diverse population. METHODS: The BACH Survey used a stratified 2-stage cluster design to randomly sample 5506 adults aged 30 to 79 from the city of Boston, Mass (2301 men, 3205 women, 1770 blacks, 1877 Hispanics, and 1859 whites). Data were obtained using interviewer and self-administered questionnaires. The presence of LUTS was defined as an American Urological Association symptom index score of 8 or above. Quality of life was assessed using the Medical Outcomes Study 12-Item Short Form Survey (SF-12), and a measure of bother was based on the interference of urinary symptoms with various activities. Analyses were weighted to the Boston population using SUDAAN version 9.0 statistical software. RESULTS: The overall prevalence of LUTS was 18.7% and increased with age (10.5% at age 30-39 years to 25.5% at age 70-79 years) but did not differ by sex or race/ethnicity. Quality of life was significantly reduced among those with LUTS, as measured by the bother of symptoms and the SF-12 component scores. Prevalence of prescription medication use for urinary symptoms was low even among participants with LUTS, with more than 90% of participants reporting no medication use. CONCLUSIONS: In this population-based, racially and ethnically diverse random sample, LUTS were common among both men and women and increased substantially with age. Lower urinary tract symptoms had a negative impact on quality of life across age, sex, and race/ethnic groups. PMID- 17130394 TI - Quantitative retinal venular caliber and risk of cardiovascular disease in older persons: the cardiovascular health study. AB - BACKGROUND: Small vessel disease may contribute to the risk of cardiovascular disease in older persons. We describe the relation of retinal vascular caliber to incident coronary heart disease (CHD) and stroke in elderly persons. METHODS: Prospective population-based cohort study composed of 1992 men and women aged 69 to 97 years living in 4 US communities. Retinal arteriolar and venular calibers were measured from retinal photographs using a computer-assisted method. Incident CHD and stroke events were ascertained using standardized methods. RESULTS: After 5 years of follow-up, there were 115 incident CHD events and 113 incident stroke events. Participants with larger retinal venular caliber had a higher incidence of CHD (11.7%; 95% confidence interval [CI], 8.7%-15.8%, vs 8.1%; 95% CI, 5.7% 11.6%), comparing largest with smallest venular caliber quartiles, and stroke (8.4%; 95% CI, 6.0-11.7, vs 5.8%; 95% CI, 3.9-8.4). At multivariable analysis, controlling for age, sex, race, arteriolar caliber, systolic and diastolic blood pressure, diabetes, glucose concentration, cigarette smoking, pack-years of smoking, and high-density-lipoprotein and low-density lipoprotein cholesterol levels, larger retinal venular caliber was associated with incident CHD (rate ratio, 3.0; 95% CI, 1.6-5.7, comparing largest with smallest venular caliber quartiles; P(trend) = .001) and incident stroke (rate ratio, 2.2; 95% CI, 1.1 4.3; P(trend) = .02). Additional adjustment for C-reactive protein and common and internal carotid artery intimal-media thickness had minimal effect on these associations. At multivariable analysis, smaller retinal arteriolar caliber was associated with incident CHD (rate ratio, 2.0; 95% CI, 1.1-3.7, comparing largest with smallest arteriolar caliber quartiles; P = .03) but not stroke (rate ratio,1.1; 95% CI, 0.5-2.2; P = .73). CONCLUSION: Larger retinal venular caliber is independently associated with risk of cardiovascular disease in elderly persons. PMID- 17130395 TI - Longitudinal study on the role of body size in premenopausal breast cancer. AB - BACKGROUND: A high body mass index (BMI) has been related to a reduced risk of breast cancer in premenopausal women. The mechanisms underlying this association have not been elucidated. METHODS: We explored whether factors affecting ovulation may explain the inverse association between BMI (calculated as weight in kilograms divided by the square of height in meters) and breast cancer in 113,130 premenopausal participants in the Nurses' Health Study II. During 1,225,520 person-years of prospective follow-up between 1989 and 2003, 1398 incident cases of invasive breast cancer were diagnosed. Weight, height, ovulatory infertility, menstrual cycle patterns, and a multitude of covariates were assessed throughout follow-up. Cox proportional hazards regression was used to compute hazard ratios and 95% confidence intervals (CIs). RESULTS: We observed a significant linear inverse trend between current BMI and breast cancer incidence (P<.001) that was not explained by menstrual cycle characteristics or infertility due to an ovulatory disorder (covariate-adjusted hazard ratio for breast cancer in women with a BMI > or =30 vs 20.0-22.4, 0.81; 95% CI, 0.68 0.96). We found BMI at age 18 years to be the strongest predictor of breast cancer incidence (covariate-adjusted hazard ratio for breast cancer in women with a BMI at age 18 years > or =27.5 vs 20.0-22.4, 0.57; 95% CI, 0.41-0.81). CONCLUSIONS: Body size during the early phases of adult life seems to be particularly important in the development of premenopausal breast cancer. Factors other than anovulation are likely to mediate the protection conferred by a high BMI. PMID- 17130396 TI - Coffee and cirrhosis: active ingredients? PMID- 17130397 TI - Frameworks for approaching prescribing at the end of life. PMID- 17130398 TI - Sex differences in the benefits of the guidelines applied in practice project: are they due to sex? PMID- 17130399 TI - Risks of erasing wrinkles: buyer beware! PMID- 17130400 TI - False-negative diffusion-weighted imaging with lateral medullary infarction. PMID- 17130401 TI - Visualizing language deterioration in progressive aphasias. PMID- 17130402 TI - A new twist on the anatomy of dystonia: the basal ganglia and the cerebellum? PMID- 17130403 TI - The optic nerve: a window into diseases of the brain? PMID- 17130404 TI - Is the Canadian health care system better for neurologic care? PMID- 17130405 TI - Selecting promising ALS therapies in clinical trials. AB - Riluzole is the only approved medication that extends survival for patients with amyotrophic lateral sclerosis (ALS). While other potential neuroprotective agents have been evaluated in randomized clinical trials, none has shown unequivocal success and none has been approved by regulatory agencies. Few symptomatic therapies have been tested in ALS. Effectiveness for drugs with modest benefit can be established only through large phase III randomized clinical trials. With numerous potential agents but limited resources, priority should be given to agents that show promise in phase II trials before proceeding to evaluation in phase III trials. In this article, we review drug development in early phase ALS trials and introduce novel designs. First, to maximize the therapeutic potential of the test medication, we need to identify the highest dose that produces a tolerable level of side effects. Second, candidate treatments should be ranked by conducting randomized selection trials between competing new treatments. The selection paradigm adopts a statistical viewpoint different from the hypothesis testing framework in conventional trials. We exemplify this approach by describing a group-sequential selection design developed for a phase II, randomized, multicenter trial of two combination treatments in patients with ALS, and illustrate the sample size reduction from a conventional trial. PMID- 17130406 TI - Behavioral features in semantic dementia vs other forms of progressive aphasias. AB - OBJECTIVE: To compare the behavioral profiles in different variants of primary progressive aphasia (PPA). METHODS: We classified 67 patients with PPA into three clinical variants: semantic dementia (SEMD), progressive nonfluent aphasia (PNFA), and logopenic progressive aphasia (LPA), and we compared the severity of behavioral dysfunction, as measured by the Neuropsychiatric Inventory, in these groups and patients with frontotemporal dementia (FTD) and Alzheimer disease (AD). RESULTS: SEMD was associated with significantly more socioemotional behavioral dysfunction than the other two variants of PPA and than AD, specifically more disinhibition, aberrant motor behavior, and eating disorders behaviors that are typical of FTD. In contrast, PNFA and LPA did not differ from each other or from AD in the type or severity of behavioral dysfunction. Behavioral abnormalities increased in severity with disease duration in SEMD, but this association was not detected in PNFA or LPA. CONCLUSIONS: Semantic dementia is associated with significantly more behavioral dysfunction than other variants of primary progressive aphasia, specifically behavioral features typical of frontotemporal dementia. PMID- 17130407 TI - Mortality in amnestic mild cognitive impairment: a prospective community study. AB - OBJECTIVE: To assess the hazard of death in persons with and without amnestic mild cognitive impairment (aMCI). METHODS: From 1987 through 2003, persons with aMCI (n = 243) and an age- and gender-matched reference group of cognitively normal persons in Olmsted County, MN, were recruited through the Mayo Clinic Alzheimer's Disease Patient Registry and followed prospectively through 2004. Survival was estimated using Kaplan-Meier survival curves, and the hazard of death for the aMCI cohort vs the reference cohort was estimated using Cox proportional hazards models. RESULTS: Over a median follow-up of 5.7 years, persons with aMCI had increased mortality (hazard ratio [HR] = 1.7; 95% CI: 1.3 to 2.3) vs reference subjects. The hazard of death by aMCI subtype was 1.5 in persons with single-domain aMCI (95% CI: 1.1 to 2.1) and 2.9 in persons with multiple-domain aMCI (95% CI: 1.9 to 4.6) vs reference subjects. Analyses restricted to aMCI cases showed an interaction between aMCI subtype and APOE epsilon4 allele status (p = 0.003). Among aMCI cases with an APOE-epsilon4 allele, there was no difference in mortality between single- and multiple-domain aMCI (HR = 1.2; 95% CI: 0.6 to 2.3). However, among aMCI cases with no APOE epsilon4 allele, the hazard of death in multiple-domain aMCI was 4.6 (95% CI: 2.3 to 9.1) vs single-domain aMCI. CONCLUSIONS: Amnestic mild cognitive impairment is associated with increased mortality, which is greater in multiple-domain aMCI than in single-domain aMCI. Mortality in aMCI subtypes may vary by APOE-epsilon4 allele status. PMID- 17130408 TI - Predominant dystonia with marked cerebellar atrophy: a rare phenotype in familial dystonia. AB - BACKGROUND: Dystonia syndromes constitute a heterogeneous group of phenotypes that may be caused by different heredodegenerative, metabolic, or genetic diseases. OBJECTIVE: To describe the characteristics of an unusual dystonia-plus phenotype associated with cerebellar atrophy. METHODS: We selected patients with predominant dystonia and cerebellar atrophy among the 861 families referred to us for genetic testing from 1992 to 2003. The main secondary heredodegenerative causes and the major genes responsible for hereditary dystonias and autosomal dominant or recessive ataxias were excluded. RESULTS: We identified 12 patients in 8 families with an unusual dystonia-plus phenotype characterized by dystonia and cerebellar atrophy on brain MRI. The mean age at onset was 27.3 +/- 11.5 years (range: 9 to 42 years) and the mean disease duration 14.7 +/- 7.7 years (range: 4 to 30). At onset, dystonia was focal or multifocal, mainly affecting vocal cords (n = 8) and upper limbs (n = 2). During the disease course spasmodic dysphonia became severe in five patients, leading to complete aphonia in two. Dystonia became generalized in five. Cerebellar ataxia was limited to unsteadiness in most patients and progressed very slowly. The paucity of clinical cerebellar signs contrasted with the marked cerebellar atrophy on brain MRI in most patients. Four families with two affected sibs support the hypothesis of an autosomal recessive disorder. However, X-linked inheritance is possible since only men were affected. CONCLUSION: We have characterized an unusual familial phenotype associating dystonia and cerebellar atrophy in 12 male patients. PMID- 17130409 TI - Driving with distraction in Parkinson disease. AB - OBJECTIVE: To assess the effects of auditory-verbal distraction on driving performance in Parkinson disease (PD). METHODS: We tested licensed, currently active drivers with mild-to-moderate PD (n = 71) and elderly controls with no neurologic disease (n = 147) on a battery of cognitive, visual, and motor tests. While they drove on a four-lane interstate freeway in an instrumented vehicle, we determined at-fault safety errors and vehicle control measures during a distracter task (Paced Auditory Serial Addition Task [PASAT]) and on an uneventful baseline segment. RESULTS: Compared with controls, drivers with PD committed more errors during both baseline and distraction, and drove slower with higher speed variability during distraction. Although the average effect of distraction on driving performance compared with baseline was not different between the groups, the drivers with PD showed a more heterogeneous response to distraction (p < 0.001): the error count increased in 28.2% of drivers with PD (vs 15.8% in controls), decreased in 16.9% (vs 3.4%), and remained stable in 54.9% (vs 80.8%). The odds of increase in safety errors due to distraction was higher in the PD group even after adjusting for baseline errors, level of engagement in PASAT, sex, and education (odds ratio [95% CI] = 2.62 [1.19 to 5.74], p = 0.016). Decreased performance on tests of cognitive flexibility, verbal memory, postural control, and increased daytime sleepiness predicted worsening of driving performance due to distraction within the PD group. CONCLUSION: The quantitative effect of an auditory-verbal distracter task on driving performance was not significantly different between Parkinson disease (PD) and control groups. However, a significantly larger subset of drivers with PD had worsening of their driving safety errors during distraction. Measures of cognition, motor function, and sleepiness predicted effects of distraction on driving performance within the PD group. PMID- 17130410 TI - Deep brain stimulation improves orthostatic regulation of patients with Parkinson disease. AB - OBJECTIVE: To evaluate whether subthalamic nucleus (STN) stimulation has an effect on the orthostatic regulation of patients with Parkinson disease (PD), we studied cardiovascular regulation during on and off phases of STN stimulation. METHODS: We examined 14 patients with PD (mean age 58.1 +/- 5.8 years, 4 women, 10 men) with bilateral STN stimulators. Patients underwent 3 minutes of head-up tilt (HUT) testing during STN stimulation and after 90 minutes interruption of stimulation. We monitored arterial blood pressure (BP), RR intervals (RRI), respiration, and skin blood flow (SBF). Baroreflex sensitivity (BRS) was assessed as the square root of the ratio of low-frequency power of RRI to the low frequency power of systolic BP for coherences above 0.5. RESULTS: During the on phase of the STN stimulation, HUT induced no BP decrease, a significant tachycardia, and a significant decrease of SBF. During the off phase of stimulation, HUT resulted in significant decreases in BPsys and RRI and only a slight SBF decrease. HUT induced no change of BRS during stimulation, but lowered BRS when the stimulator was off (p < 0.05). CONCLUSIONS: STN stimulation of patients with PD increases peripheral vasoconstriction and BRS and stabilizes BP, thereby improving postural hypotension in patients with PD. The results indicate that STN stimulation not only alleviates motor deficits but also influences autonomic regulation in patients with PD. PMID- 17130411 TI - Pregabalin in central neuropathic pain associated with spinal cord injury: a placebo-controlled trial. AB - OBJECTIVE: To evaluate pregabalin in central neuropathic pain associated with spinal cord injury. METHODS: A 12-week, multicenter study of patients randomized to either flexible-dose pregabalin 150 to 600 mg/day (n = 70) or placebo (n = 67), administered BID. Patients were allowed to remain on existing, stable pain therapy. The primary efficacy variable was the endpoint mean pain score, derived from patients' last 7 days daily pain diary entries. Key secondary endpoints included pain responder rates, the SF-MPQ, sleep interference, mood, and the patient global measure of change. RESULTS: The mean baseline pain score was 6.54 in the pregabalin group and 6.73 in the placebo group. The mean endpoint pain score was lower in the pregabalin group (4.62) than the placebo group (6.27; p < 0.001), with efficacy observed as early as week 1 and maintained for the duration of the study. The average pregabalin dose after the 3-week stabilization phase was 460 mg/day. Pregabalin was significantly superior to placebo in endpoint assessments on the SF-MPQ. The > or =30% and > or =50% pain responder rates were higher with pregabalin than placebo (p < 0.05). Pregabalin was associated with improvements in disturbed sleep (p < 0.001) and anxiety (p < 0.05), and more patients reported global improvement at endpoint in the pregabalin group (p < 0.001). Mild or moderate, typically transient, somnolence and dizziness were the most common adverse events. CONCLUSIONS: Pregabalin 150 to 600 mg/day was effective in relieving central neuropathic pain, improving sleep, anxiety, and overall patient status in patients with spinal cord injury. PMID- 17130412 TI - Left size distortion (hyperschematia) after right brain damage. AB - OBJECTIVE: To quantitate a size distortion involving the side of space contralateral to the lesion (contralesional) in two right-brain-damaged patients. METHODS: We studied two right-brain-damaged patients with lesions sparing the occipital lobe and a mild left neglect on target cancellation or line bisection. The lesions involved the temporoparietal region (Patient 1) and the basal ganglia and the insula (Patients 1 and 2). Patients were given drawing tasks and tasks requiring perceptual and visuomotor judgments of horizontal extent. RESULTS: In drawing objects such as a daisy both from memory and by copying, patients exhibited a disproportionate enlargement of the left-hand side of objects and added more left-sided petals to the drawn daisy. This pathologic behavior persisted when the patients were blindfolded and was likely to reflect a perceptual, rather than premotor, size distortion. In a task requiring the perceptual matching of two rectangles, patients underestimated the left-sided stimulus. In a visuomotor task requiring the reproduction of the horizontal extent of a segment, patients exhibited a hyperextension, when a leftward movement was required. CONCLUSIONS: We showed a disordered representation of extrapersonal space, possibly involving a contralesional relaxation of the spatial medium. The deficit does not arise at the level of retinotopic coordinate frames and is independent of unilateral spatial neglect. PMID- 17130413 TI - Incidence and outcome of cervical artery dissection: a population-based study. AB - BACKGROUND: Incidence rates for internal carotid artery dissection (ICAD) have been reported to be 2.6 to 2.9 per 100,000, but reliable epidemiologic data for vertebral artery dissection (VAD) are not available. OBJECTIVE: To determine the incidence rate of cervical artery dissection (CAD) in a defined population. METHODS: With IRB approval, we used the medical record linkage system of the Rochester Epidemiology Project to identify all patients diagnosed with spontaneous ICAD and VAD for the period of 1987-2003 in Olmsted County, MN. RESULTS: Of 48 patients with CAD, there were 32 patients with ICAD and 18 patients with VAD. In Olmsted County, the average annual incidence rate for ICAD was 1.72 per 100,000 population (95% CI, 1.13 to 2.32) and for VAD 0.97 per 100,000 population (95% CI, 0.52 to 1.4). The average annual incidence rate for CAD was 2.6 per 100,000 population (95% CI, 1.86 to 3.33). The most frequently encountered symptoms in CAD were head or neck pain (80%), cerebral ischemia (TIA or infarct) (56%), and Horner syndrome (25%). Good outcome (defined as modified Rankin score of 0 to 2) was seen in 92% of patients. No recurrence of dissection was observed during a mean 7.8 years of follow-up. CONCLUSIONS: Internal carotid artery dissection was detected approximately twice as frequently as vertebral artery dissection in the overall study, but in the latter half of the study period, vertebral artery and internal carotid artery dissection incidence rates were equivalent. The majority of cervical artery dissection patients in the community have excellent outcome, and contrary to many tertiary referral series, re-dissection is rare. PMID- 17130414 TI - Relationship between language lateralization and handedness in left-hemispheric partial epilepsy. AB - OBJECTIVE: To investigate the relationship between language lateralization and handedness in patients with epilepsy and a left-sided seizure focus and in healthy control subjects. METHODS: We recruited a consecutive series of 74 patients and 70 control subjects. Functional MRI, using a noun-verb generation task, was performed to establish the language laterality index (LI). Handedness was quantified using the Edinburgh Handedness Inventory. RESULTS: Patients showed a shift toward atypical language lateralization (0.43 +/- 0.47; controls 0.57 +/- 034; p < or = 0.05) and left-handedness (55 +/- 57; controls 74 +/- 39; p < or = 0.05). The LI and handedness were correlated in patients (r = 0.54; F = 25.9; p < 0.001) but not in control subjects (r = 0.1; F = 0.64; NS). A combination of left handedness and atypical LI was more frequent in patients (12%) than control subjects (0%; p < or = 0.05). Crossed hemispheric specialization (e.g., right handedness associated with atypical LI) was equally frequent in patients (20%) and control subjects (16%; NS). CONCLUSION: In epilepsy patients with a left sided seizure focus, language lateralization is correlated to handedness. The increased frequency of left-handedness and associated atypical language lateralization is most likely related to the left-hemispheric seizure focus, influencing hemispheric specialization for both domains. PMID- 17130415 TI - Abnormal orbitofrontal development due to prematurity. AB - OBJECTIVE: To investigate the effects of prematurity on sulcal formation. METHODS: We evaluated the depth and volume of the primary olfactory sulcus (developed at 16 weeks' gestation) and the secondary orbital sulci (which start to develop at 28 weeks' gestation) in a sample of 22 adolescents with history of very-preterm birth (VPTB). We compared this preterm sample with a sample of subjects born at term and matched by age, gender, and sociocultural status. The Anatomist/BrainVISA 3.0.1 package was used to identify and quantify the sulci. In addition, voxel-based morphometry (VBM) was used to analyze possible reductions of gray and white matter in the orbitofrontal area. RESULTS: Compared with controls, we found a significant reduction in the secondary sulci depth but not in the primary sulcus in the VPTB. VBM analysis showed reduced gray-matter volume in VPTB in the orbital region. CONCLUSIONS: Premature birth affects cerebral gyrification, and this impairment is not reversible during childhood. Identification of the specific factors involved in abnormal brain maturation may lead to effective interventions. PMID- 17130416 TI - Mitochondrial disease criteria: diagnostic applications in children. AB - BACKGROUND: Based on a previous prospective clinical and biochemical study, a consensus mitochondrial disease scoring system was established to facilitate the diagnosis in patients with a suspected mitochondrial disorder. OBJECTIVE: To evaluate the specificity of the diagnostic system, we applied the mitochondrial disease score in 61 children with a multisystem disease and a suspected oxidative phosphorylation disorder who underwent a muscle biopsy and were consecutively diagnosed with a genetic mutation. METHODS: We evaluated data of 44 children diagnosed with a disorder in oxidative phosphorylation, carrying a mutation in the mitochondrial or nuclear DNA. We compared them with 17 children who, based on the clinical and metabolic features, also had a muscle biopsy but were finally diagnosed with a nonmitochondrial multisystem disorder by further genetic analysis. RESULTS: All children with a genetically established diagnosis of a primary oxidative phosphorylation disorder had a mitochondrial disease score above 6 (probable mitochondrial disorder), and 73% of the children had a score above 8 (definite mitochondrial disorder) at evaluation of the muscle biopsy. In the nonmitochondrial multisystem disorder group, the score was significantly lower, and no patients reached a score comparable with a definite respiratory chain disorder. CONCLUSIONS: The mitochondrial disease criteria system has a high specificity to distinguish between mitochondrial and other multisystem disorders. The method could also be applied in children with a suspected mitochondrial disorder, prior to performing a muscle biopsy. PMID- 17130417 TI - The value of database controls in pilot or futility studies in ALS. AB - OBJECTIVE: To evaluate the use and reliability of database controls in place of a placebo group in pilot or "futility" ALS trials. METHODS: We compared the rates of disease progression in the placebo arm of the clinical phase III US Insulin like Growth Factor-I Trial (n = 90) with the rates of disease progression of 207 patients with ALS selected from 1,600 ALS database patients using the same inclusion criteria. RESULTS: The mean rates of change in the Appel ALS (AALS) score were nearly identical in the placebo group (4.70 points/month) and in the database group (4.79 points/month). In addition, there was no significant difference in the median time to achieving a 20-point progression in AALS score: 143 days for database match vs 146 days for the placebo group (log rank p = 0.88). Furthermore, in the multivariate Cox analysis, both the rate at which the disease had progressed prior to first exam (preslope) (p < 0.001) and first exam AALS total score (p = 0.01) were shown to be covariates of subsequent rate of disease progression. CONCLUSION: The similarity in disease progression between placebo arm of clinical phase III trial and matched database group suggests the value of historical databases in futility trials. However, the proposed study design requires appropriate matching of study patients with database controls. Based on our results, we suggest matching by stage of the disease and rate of clinical decline in a contemporaneous ALS population. PMID- 17130418 TI - Elevated serum angiogenin levels in ALS. AB - BACKGROUND: The role of hypoxia responsive genes in the pathogenesis of ALS was first suggested when deletions of the hypoxia-responsive element of vascular endothelial growth factor (VEGF) promoter caused a motor neuron disease phenotype in mice. The discovery of ALS-associated mutations in ANG, a hypoxia responsive gene coding for the protein angiogenin, has further supported this pathogenic mechanism in human ALS. In endothelium, angiogenin can regulate expression of VEGF. To date, the patterns of serum angiogenin expression among patients with ALS have not been assessed. METHODS: Serum angiogenin and VEGF levels were quantified at diagnosis in 79 patients with definite or probable ALS and 72 healthy controls, using a quantitative sandwich enzyme-linked immunoassay. RESULTS: Patients with ALS exhibited higher serum angiogenin (p = 0.006) but not VEGF (p = 0.55) levels than matched control subjects. Subgroup analysis showed a greater elevation in angiogenin levels for spinal- (p < 0.001) than bulbar- (p = 0.11) onset ALS vs controls. At 12 months, angiogenin levels remained elevated. No correlation was noted between angiogenin and VEGF levels (r = -0.08, p = 0.49) in ALS patient serum. CONCLUSION: These data suggest a modest elevation in serum angiogenin in ALS at diagnosis. Further investigation will be required to assess the utility of serum angiogenin as a biomarker for ALS and as a predictor of disease progression. PMID- 17130419 TI - Relationship of bacterial strains to clinical syndromes of Campylobacter associated neuropathies. AB - BACKGROUND: Clinical and serologic studies suggest that Guillain-Barre syndrome (GBS) and atypical GBS with preserved muscle stretch reflexes (MSRs) form a continuous spectrum as well as do Fisher syndrome (FS), FS/GBS overlap, Bickerstaff brainstem encephalitis (BBE), BBE/GBS overlap, acute ophthalmoparesis (AO), ataxic GBS, and acute oropharyngeal palsy. OBJECTIVE: To clarify the spectrum of neurologic disorders that occur subsequent to Campylobacter jejuni enteritis. METHODS: We recruited patients with various neurologic conditions and from whom C jejuni was isolated. Bacterial features were investigated. RESULTS: Diagnoses for the patients from whom C jejuni was isolated were GBS (n = 90), FS (n = 22), MSR-preserved GBS (n = 10), FS/GBS (n = 6), BBE (n = 1), BBE/GBS (n = 2), AO (n = 3), ataxic GBS (n = 1), and acute oropharyngeal palsy (n = 3). Isolates from MSR-preserved GBS were similar to those of GBS in serotype (HS:19), genotype (lipo-oligosaccharide [LOS] locus class A or B, cst-II genotype [Thr51]), and GM1 or GD1a epitope expression on LOS. FS/GBS overlap, BBE, BBE/GBS overlap, AO, ataxic GBS, and acute oropharyngeal palsy isolates were similar to those of FS in serotype (HS:2 or HS:4-complex), genotype (LOS locus class A or B, cst-II genotype [Asn51]), and GQ1b epitope expression on LOS. CONCLUSIONS: The bacterial findings support the proposal that Guillain-Barre syndrome (GBS) and muscle stretch reflex-preserved GBS comprise a continuous spectrum as well as do Fisher syndrome (FS), FS/GBS overlap, Bickerstaff brainstem encephalitis (BBE), BBE/GBS overlap, acute ophthalmoparesis, ataxic GBS, and acute oropharyngeal palsy. PMID- 17130420 TI - Acute hypothalamic stimulation and ongoing cluster headache attacks. AB - Long-term hypothalamic stimulation is effective in improving drug-resistant chronic cluster headache (CH). We assessed acute hypothalamic stimulation to resolve ongoing CH attacks in 16 patients implanted to prevent chronic CH, investigating 136 attacks. A pain intensity reduction of > or =50% occurred in 25 of 108 evaluable attacks (23.1%). Acute hypothalamic stimulation is not effective in resolving ongoing CH attacks, suggesting that hypothalamic stimulation acts by complex mechanisms in CH prevention. PMID- 17130421 TI - Prediction of thrombolytic efficacy in acute ischemic stroke using thin-section noncontrast CT. AB - Thin-section noncontrast CT (NCT) can detect thrombi in large arteries and can provide a measure of thrombus composition based on Hounsfield Units (HU). A study using polyethylene tubes as a model of intracranial arteries concluded that the HUs of platelet-rich thrombi are lower than those of erythrocyte-rich thrombi. Thrombus HUs were measured by NCT in 34 patients with acute ischemic stroke before thrombolysis. Thrombi with lower HU counts were resistant to thrombolytics. PMID- 17130422 TI - Reduction of optic nerve fibers in patients with Alzheimer disease identified by laser imaging. AB - In this case-control study, we compared the optic nerves (ONs) by clinical examination and scanning laser ophthalmoscopy (SLO) of 40 patients with Alzheimer disease (AD) and 50 controls. There was a reduction in the number of ON fibers in patients with AD, with a threefold greater odds ratio for a larger optic cup-to disc ratio in patients with AD. PMID- 17130423 TI - Severe botulism after focal injection of botulinum toxin. AB - We report a 34-year-old woman who developed clinical botulism after the cosmetic use of an unapproved botulinum toxin type A. Electrophysiologic findings demonstrated complete denervation with complete electrical silence. She had a lengthy recovery but was able to ambulate by discharge. PMID- 17130424 TI - Strong genetic evidence for association of TOR1A/TOR1B with idiopathic dystonia. AB - Recently, association of a TOR1A(DYT1)/TOR1B risk haplotype with common forms of idiopathic dystonia has been reported in the Icelandic population. Here we report a strong association of two single nucleotide polymorphisms within or in close proximity to the TOR1A 3'UTR, with the lowest p value being 0.000008, in a larger cohort of German and Austrian patients with predominantly focal sporadic dystonia. PMID- 17130425 TI - Tumor suppressor in lung cancer-1 (TSLC1) functions as a glioma tumor suppressor. AB - Tumor suppressor in lung cancer-1 (TSLC1) loss is common in many human cancers, including meningioma. In this study, we demonstrate that TSLC1 protein and RNA expression is lost in 60% to 65% of high-grade gliomas, and that TSLC1 reintroduction into glioma cells results in growth suppression. Moreover, Tslc1 loss in mice results in increased astrocyte proliferation in vivo and in vitro. These data indicate that TSLC1 functions as a glioma tumor suppressor. PMID- 17130426 TI - Exploded drawing in posterior cortical atrophy. PMID- 17130427 TI - Diminished somatostatin gene expression in individuals with HIV and major depressive disorder. AB - In analyzing the expression of 15 candidate genes for HIV encephalitis (HIVE) by the presence or absence of major depressive disorder (MDD), we noted significant reductions in the expression of four cytoskeletal genes and somatostatin. Whereas disruption of cytoskeletal genes has been noted in HIVE, dysregulation of somatostatin has not, indicating that dysregulation of somatostatin is part of the molecular pathologic process of MDD in the setting of HIV. PMID- 17130428 TI - Low recurrence rate of vestibular neuritis: a long-term follow-up. AB - We examined 103 patients with vestibular neuritis (VN) in a follow-up study (5.7 to 20.5 years, mean 9.8 years). Two patients (1.9%) had developed a second occurrence of VN 29 to 39 months after the first. VN affected the contralateral ear in both and caused less severe distressing vertigo and postural imbalance. Unlike Bell's palsy and sudden hearing loss, a relapse in the same ear did not occur. PMID- 17130429 TI - Efficacy of carbamazepine and valproate as monotherapy for early epilepsy and single seizures. AB - In the Multi-Centre Study of Early Epilepsy and Single Seizures (MESS), patients were randomly allocated to immediate or delayed antiepileptic drug treatment. For time to first seizure recurrence, MESS provides strong evidence of an effect for carbamazepine as monotherapy but mixed evidence of an effect for valproate. PMID- 17130430 TI - Levetiracetam in pregnancy: preliminary experience from the UK Epilepsy and Pregnancy Register. AB - It is not known whether the antiepileptic drug (AED) levetiracetam can be used safely in human pregnancy. As part of a study to determine the risks of major congenital malformations (MCMs) for infants exposed to AEDs in utero, we identified all cases exposed to levetiracetam. Three of 117 exposed pregnancies had an MCM (2.7%; 95% CI 0.9% to 7.7%); all 3 were exposed to other AEDs. PMID- 17130431 TI - Blockade of chemokine signaling in patients with multiple sclerosis. AB - We assessed the safety and efficacy of orally administered CC chemokine receptor 1 (CCR1) antagonist in 105 patients with relapsing/remitting MS (RRMS) in a 16 week, randomized, double-blind, placebo-controlled trial. The primary endpoint was the cumulative number of newly active lesions on serial MRI scans. Other MRI, immunologic, and clinical outcomes were also explored. No significant treatment difference was observed for any tested MRI variable. CCR1 does not contribute to initial leukocyte infiltration in RRMS. PMID- 17130432 TI - Upper limb tremor induced by peripheral nerve injury. AB - We report a patient with proximal right upper limb tremor, secondary to direct peripheral nerve lesion caused by prior thoracic surgery. Electromyography demonstrated neurogenic abnormalities and tremor in muscles innervated by the thoracodorsal and long thoracic nerves. Somatosensory evoked potentials, transcranial magnetic stimulation, and MRI of the cervical and thoracic spine were normal. Tremor persisted in REM and non-REM sleep. These findings suggest a peripheral generator. PMID- 17130433 TI - Pregnancy and birth outcomes in women with facioscapulohumeral muscular dystrophy. AB - Obstetric risk in facioscapulohumeral muscular dystrophy (FSHD) is not known. We surveyed 38 women with FSHD reporting 105 gestations and 78 live births. Review of medical records showed that pregnancy outcomes were generally favorable. The rates for low birth weight and total operative deliveries were statistically higher than the national rates in the general population. Worsening of FSHD was reported in 24% of gestations and did not usually resolve after delivery. PMID- 17130434 TI - Locked-in syndrome resulting from bilateral cerebral peduncle infarctions. PMID- 17130435 TI - An adult case of leukoencephalopathy with intracranial calcifications and cysts. AB - We describe a 44-year-old woman with progressive headache, ataxia, and seizures in association with multifocal cerebral and cerebellar leukoencephalopathy, intracranial calcifications, and cysts. The cause of death was intracerebellar hemorrhage while taking warfarin. Pathologic features on biopsy included angiomatous-like blood vessels, intense gliosis, and Rosenthal fiber formation in the white matter. Genetic analyses did not identify any significant mutations in two candidate genes. PMID- 17130436 TI - Multiple myeloma presenting with acute disseminated encephalomyelitis: causal or chance link? PMID- 17130437 TI - Motor neuron disorder simulating ALS induced by chronic inhalation of pyrethroid insecticides. PMID- 17130438 TI - 3-Methylglutaconic aciduria type I causes leukoencephalopathy of adult onset. PMID- 17130439 TI - Sighs too deep for words. PMID- 17130440 TI - Treadmill training after spinal cord injury: good but not better. PMID- 17130441 TI - Weight-supported treadmill vs over-ground training for walking after acute incomplete SCI. PMID- 17130442 TI - Neuropsychological deficits in long-term frequent cannabis users. PMID- 17130443 TI - Disequilibrium and association in barley: thinking outside the glass. PMID- 17130444 TI - Revealing the ghost in the machine: using spectral analysis to understand the influence of noise on population dynamics. PMID- 17130445 TI - Four-dimensional ultrafast electron microscopy of phase transitions. AB - Reported here is direct imaging (and diffraction) by using 4D ultrafast electron microscopy (UEM) with combined spatial and temporal resolutions. In the first phase of UEM, it was possible to obtain snapshot images by using timed, single electron packets; each packet is free of space-charge effects. Here, we demonstrate the ability to obtain sequences of snapshots ("movies") with atomic scale spatial resolution and ultrashort temporal resolution. Specifically, it is shown that ultrafast metal-insulator phase transitions can be studied with these achieved spatial and temporal resolutions. The diffraction (atomic scale) and images (nanometer scale) we obtained manifest the structural phase transition with its characteristic hysteresis, and the time scale involved (100 fs) is now studied by directly monitoring coordinates of the atoms themselves. PMID- 17130446 TI - Unliganded and hormone-bound glucocorticoid receptors interact with distinct hydrophobic sites in the Hsp90 C-terminal domain. AB - Unlike most chaperones, heat-shock protein 90 (Hsp90) interacts with a select group of "client proteins" that regulate essential biological processes. Little is known about how Hsp90 recognizes and binds these proteins. The glucocorticoid receptor (GR) is a well characterized Hsp90 client protein, whose hormone binding, nuclear-cytoplasmic trafficking, and transcriptional activity are regulated by Hsp90. Here, we provide evidence that unliganded and hormone-bound GR interact with two distinct, solvent-exposed hydrophobic sites in the Hsp90 C terminal domain that contain the sequences "MxxIM" (HM10) and "L/MxxIL" (HM9). Our results indicate that binding of Hsp90 HM10 to unliganded GR stabilizes the unliganded ligand-binding pocket of GR indirectly by promoting an intramolecular interaction between the C-terminal alpha-helix (H12) and a solvent-exposed hydrophobic groove in the GR ligand binding domain. In the presence of hormone, Hsp90 appears to bind the hydrophobic groove of GR directly by mimicking the interactions of GR with transcriptional coactivators. The identified interactions provide insights into the mechanisms that enable Hsp90 to regulate the activity of both unliganded and hormone-bound GR and to sharpen the cellular response to hormone. PMID- 17130447 TI - Prevention and reversal of cardiac hypertrophy by soluble epoxide hydrolase inhibitors. AB - Sustained cardiac hypertrophy represents one of the most common causes leading to cardiac failure. There is emerging evidence to implicate the involvement of NF kappaB in the development of cardiac hypertrophy. However, several critical questions remain unanswered. We tested the use of soluble epoxide hydrolase (sEH) inhibitors as a means to enhance the biological activities of epoxyeicosatrienoic acids (EETs) to treat cardiac hypertrophy. sEH catalyzes the conversion of EETs to form the corresponding dihydroxyeicosatrienoic acids. Previous data have suggested that EETs may inhibit the activation of NF-kappaB-mediated gene transcription. We directly demonstrate the beneficial effects of several potent sEH inhibitors (sEHIs) in cardiac hypertrophy. Specifically, we show that sEHIs can prevent the development of cardiac hypertrophy using a murine model of pressure-induced cardiac hypertrophy. In addition, sEHIs reverse the preestablished cardiac hypertrophy caused by chronic pressure overload. We further demonstrate that these compounds potently block the NF-kappaB activation in cardiac myocytes. Moreover, by using in vivo electrophysiologic recordings, our study shows a beneficial effect of the compounds in the prevention of cardiac arrhythmias that occur in association with cardiac hypertrophy. We conclude that the use of sEHIs to increase the level of the endogenous lipid epoxides such as EETs may represent a viable and completely unexplored avenue to reduce cardiac hypertrophy by blocking NF-kappaB activation. PMID- 17130448 TI - Angiopoietin-like 4 prevents metastasis through inhibition of vascular permeability and tumor cell motility and invasiveness. AB - Angiopoietin-like 4 (ANGPTL4), a secreted protein of the angiopoietin-like family, is induced by hypoxia in both tumor and endothelial cells as well as in hypoxic perinecrotic areas of numerous cancers. Here, we investigated whether ANGPTL4 might affect tumor growth as well as metastasis. Metastatic 3LL cells were therefore xenografted into control mice and mice in which ANGPTL4 was expressed by using in vivo DNA electrotransfer. Whereas primary tumors grew at a similar rate in both groups, 3LL cells metastasized less efficiently to the lungs of mice that expressed ANGPTL4. Fewer 3LL emboli were observed in primary tumors, suggesting that intravasation of 3LL cells was inhibited by ANGPTL4. Furthermore, melanoma B16F0 cells injected into the retro-orbital sinus also metastasized less efficiently in mice expressing ANGPTL4. Although B16F0 cells were observed in lung vessels, they rarely invaded the parenchyma, suggesting that ANGPTL4 affects extravasation. In addition, recombinant B16F0 cells that overexpress ANGPTL4 were generated, showing a lower capacity for in vitro migration, invasion, and adhesion than control cells. Expression of ANGPTL4 induced reorganization of the actin cytoskeleton through inhibition of actin stress fiber formation and vinculin localization at focal contacts. Together, these results show that ANGPTL4, through its action on both vascular and tumor compartments, prevents the metastatic process by inhibiting vascular activity as well as tumor cell motility and invasiveness. PMID- 17130449 TI - A driving and coupling "Pac-Man" mechanism for chromosome poleward translocation in anaphase A. AB - During mitosis, chromatid harnesses its kinetochore translocation at the depolymerizing microtubule ends for its poleward movement in anaphase A. The force generation mechanism for such movement remains unknown. Analysis of the current experimental results shows that the bending energy release from the bound tubulin subunits alone cannot provide sufficient driving force. Additional contribution from effective electrostatic attractions between the kinetochore and the microtubule is needed for kinetochore translocation. Interestingly, as the kinetochore moves to inside the microtubule, the microtubule tip is free to bend outward so that the instantaneous distance between the kinetochore and the microtubule tip is much closer than the rest of the microtubule. This close contact yields much larger electrostatic attraction than that from the rest of the microtubule under physiological ionic conditions. As a result, the effective electrostatic interaction hinders the further kinetochore poleward translocation until the microtubule tip dissociates. Thus, the kinetochore translocation is strongly coupled at the depolymerizing microtubule end. This driving-coupling mechanism indicates that the kinetochore velocity is largely controlled by the microtubule dissociation rate, which explains the insensitivity of kinetochore velocity to its viscous drag and the large redundancy in its stalling force. PMID- 17130450 TI - Sharing of T cell receptors in antigen-specific responses is driven by convergent recombination. AB - Public responses where identical T cell receptors (TCRs) are clonally dominant and shared between different individuals are a common characteristic of CD8(+) T cell-mediated immunity. Focusing on TCR sharing, we analyzed approximately 3,400 TCR beta chains (TCRbetas) from mouse CD8(+) T cells responding to the influenza A virus D(b)NP(366) and D(b)PA(224) epitopes. Both the "public" D(b)NP(366) specific and "private" D(b)PA(224)-specific TCR repertoires contain a high proportion ( approximately 36%) of shared TCRbetas, although the numbers of mice sharing TCRbetas in each repertoire varies greatly. Sharing of both the TCRbeta amino acid and TCRbeta nucleotide sequence was negatively correlated with the prevalence of random nucleotide additions in the sequence. However, the extent of TCRbeta amino acid sequence sharing among mice was strongly correlated with the level of diversity in the encoding nucleotide sequences, suggesting that a key feature of public TCRs is that they can be made in a variety of ways. Using a computer simulation of random V(D)J recombination, we estimated the relative production frequencies and variety of production mechanisms for TCRbeta sequences and found strong correlations with the sharing of both TCRbeta amino acid sequences and TCRbeta nucleotide sequences. The overall conclusion is that "convergent recombination," rather than a bias in recombination or subsequent selection, provides the mechanistic basis for TCR sharing between individuals responding to identical peptide plus MHC class I glycoprotein complexes. PMID- 17130451 TI - Klf4 and corticosteroids activate an overlapping set of transcriptional targets to accelerate in utero epidermal barrier acquisition. AB - Premature infants are at an increased risk for infections and dehydration because of incomplete development of the epidermis, which attains its essential function as a barrier only during the last stages of in utero development. When a premature birth is anticipated, antenatal corticosteroids are administered to accelerate lung epithelium differentiation. One pleiotropic, but beneficial, effect of antenatal corticosteroids is acceleration of skin barrier establishment by an unknown mechanism. In mice, the transcription factor Klf4 is both necessary and sufficient, within a developmental field of competence, to establish this skin barrier, as demonstrated by targeted ablation and transgenic expression of Klf4, respectively. Here, we report that Klf4 and corticosteroid treatment coordinately accelerate barrier acquisition in vivo. Transcriptional profiling reveals that the genes regulated by corticosteroids and Klf4 during the critical window of epidermal development significantly overlap. KLF4 activates the proximal promoters of a significant subset of these genes. Dissecting the intersection of the genetic and pharmacological pathways, regulated by KLF4 and corticosteroids, respectively, leads to a mechanistic understanding of the normal process of epidermal development in utero. PMID- 17130452 TI - Two different domains of the luciferase gene in the heterotrophic dinoflagellate Noctiluca scintillans occur as two separate genes in photosynthetic species. AB - Noctiluca scintillans, a heterotrophic unarmored unicellular bioluminescent dinoflagellate, occurs widely in the oceans, often as a bloom. Molecular phylogenetic analysis based on 18S ribosomal DNA sequences consistently has placed this species on the basal branch of dinoflagellates. Here, we report that the structural organization of its luciferase gene is strikingly different from that of the seven luminous species previously characterized, all of which are photosynthetic. The Noctiluca gene codes for a polypeptide that consists of two distinct but contiguous domains. One, which is located in the N-terminal portion, is shorter than but similar in sequence to the individual domains of the three domain luciferases found in all other luminous dinoflagellates studied. The other, situated in the C-terminal part, has sequence similarity to the luciferin binding protein of the luminous dinoflagellate Lingulodinium polyedrum, encoded there by a separate gene. Western analysis shows that the native protein has the same size (approximately 100 kDa) as the heterologously expressed polypeptide, indicating that it is not a polyprotein. Thus, sequences found in two proteins in the L. polyedrum bioluminescence system are present in a single polypeptide in Noctiluca. PMID- 17130453 TI - Sensory integration does not lead to sensory calibration. AB - One generally has the impression that one feels one's hand at the same location as one sees it. However, because our brain deals with possibly conflicting visual and proprioceptive information about hand position by combining it into an optimal estimate of the hand's location, mutual calibration is not necessary to achieve such a coherent percept. Does sensory integration nevertheless entail sensory calibration? We asked subjects to move their hand between visual targets. Blocks of trials without any visual feedback about their hand's position were alternated with blocks with veridical visual feedback. Whenever vision was removed, individual subjects' hands slowly drifted toward the same position to which they had drifted on previous blocks without visual feedback. The time course of the observed drift depended in a predictable manner (assuming optimal sensory combination) on the variable errors in the blocks with and without visual feedback. We conclude that the optimal use of unaligned sensory information, rather than changes within either of the senses or an accumulation of execution errors, is the cause of the frequently observed movement drift. The conclusion that seeing one's hand does not lead to an alignment between vision and proprioception has important consequences for the interpretation of previous work on visuomotor adaptation. PMID- 17130454 TI - From the Academy: Colloquium perspective. Toward cropping systems that enhance productivity and sustainability. AB - The defining features of any cropping system are (i) the crop rotation and (ii) the kind or intensity of tillage. The trend worldwide starting in the late 20th century has been (i) to specialize competitively in the production of two, three, a single, or closely related crops such as different market classes of wheat and barley, and (ii) to use direct seeding, also known as no-till, to cut costs and save soil, time, and fuel. The availability of glyphosate- and insect-resistant varieties of soybeans, corn, cotton, and canola has helped greatly to address weed and insect pest pressures favored by direct seeding these crops. However, little has been done through genetics and breeding to address diseases caused by residue- and soil-inhabiting pathogens that remain major obstacles to wider adoption of these potentially more productive and sustainable systems. Instead, the gains have been due largely to innovations in management, including enhancement of root defense by antibiotic-producing rhizosphere-inhabiting bacteria inhibitory to root pathogens. Historically, new varieties have facilitated wider adoption of new management, and changes in management have facilitated wider adoption of new varieties. Although actual yields may be lower in direct-seed compared with conventional cropping systems, largely due to diseases, the yield potential is higher because of more available water and increases in soil organic matter. Achieving the full production potential of these more-sustainable cropping systems must now await the development of varieties adapted to or resistant to the hazards shown to account for the yield depressions associated with direct seeding. PMID- 17130455 TI - Intracellular protein interaction mapping with FRET hybrids. AB - A quantitative methodology was developed to identify protein interactions in a broad range of cell types by using FRET between fluorescent proteins. Genetic fusions of a target receptor to a FRET acceptor and a large library of candidate peptide ligands to a FRET donor enabled high-throughput optical screening for optimal interaction partners in the cytoplasm of Escherichia coli. Flow cytometric screening identified a panel of peptide ligands capable of recognizing the target receptors in the intracellular environment. For both SH3 and PDZ domain-type target receptors, physiologically meaningful consensus sequences were apparent among the isolated ligands. The relative dissociation constants of interacting partners could be measured directly by using a dilution series of cell lysates containing FRET hybrids, providing a previously undescribed high throughput approach to rank the affinity of many interaction partners. FRET hybrid interaction screening provides a powerful tool to discover protein ligands in the cellular context with potential applications to a wide variety of eukaryotic cell types. PMID- 17130456 TI - Neutron diffraction studies of Escherichia coli dihydrofolate reductase complexed with methotrexate. AB - Hydrogen atoms play a central role in many biochemical processes yet are difficult to visualize by x-ray crystallography. Spallation neutron sources provide a new arena for protein crystallography with TOF measurements enhancing data collection efficiency and allowing hydrogen atoms to be located in smaller crystals of larger biological macromolecules. Here we report a 2.2-A resolution neutron structure of Escherichia coli dihydrofolate reductase (DHFR) in complex with methotrexate (MTX). Neutron data were collected on a 0.3-mm(3) D(2)O-soaked crystal at the Los Alamos Neutron Scattering Center. This study provides an example of using spallation neutrons to study protein dynamics, to identify protonation states directly from nuclear density maps, and to analyze solvent structure. Our structure reveals that the occluded loop conformation [monomer (mon.) A] of the DHFR.MTX complex undergoes greater H/D exchange compared with the closed-loop conformer (mon. B), partly because the Met-20 and beta(F-G) loops readily exchange in mon. A. The eight-stranded beta sheet of both DHFR molecules resists H/D exchange more than the helices and loops. However, the C-terminal strand, betaH, in mon. A is almost fully exchanged. Several D(2)Os form hydrogen bonds with exchanged amides. At the active site, the N1 atom of MTX is protonated and thus charged when bound to DHFR. Several D(2)Os are observed at hydrophobic surfaces, including two pockets near the MTX-binding site. A previously unidentified D(2)O hydrogen bonds with the catalytic D27 in mon. B, stabilizing its negative charge. PMID- 17130457 TI - A BTB/POZ protein, NAC-1, is related to tumor recurrence and is essential for tumor growth and survival. AB - Recent studies have suggested an oncogenic role of the BTB/POZ-domain genes in hematopoietic malignancy. The aim of this study is to identify and characterize BTB/POZ-domain genes in the development of human epithelial cancers, i.e., carcinomas. In this study, we focused on ovarian carcinoma and analyzed gene expression levels using the serial analysis of gene expression (SAGE) data in all 130 deduced BTB/POZ genes. Our analysis reveals that NAC-1 is significantly overexpressed in ovarian serous carcinomas and several other types of carcinomas. Immunohistochemistry studies in ovarian serous carcinomas demonstrate that NAC-1 is localized in discrete nuclear bodies (tentatively named NAC-1 bodies), and the levels of NAC-1 expression correlate with tumor recurrence. Furthermore, intense NAC-1 immunoreactivity in primary tumors predicts early recurrence in ovarian cancer. Both coimmunoprecipitation and double immunofluorescence staining demonstrate that NAC-1 molecules homooligomerize through the BTB/POZ domain. Induced expression of the NAC-1 mutant containing only the BTB/POZ domain disrupts NAC-1 bodies, prevents tumor formation, and promotes tumor cell apoptosis in established tumors in a mouse xenograft model. Overexpression of full-length NAC-1 enhanced tumorigenicity of ovarian surface epithelial cells and NIH 3T3 cells in athymic nu/nu mice. In summary, NAC-1 is a tumor recurrence associated gene with oncogenic potential, and the interaction between BTB/POZ domains of NAC-1 proteins is critical to form the discrete NAC-1 nuclear bodies and essential for tumor cell proliferation and survival. PMID- 17130458 TI - Empirical Bayes hierarchical models for regularizing maximum likelihood estimation in the matrix Gaussian Procrustes problem. AB - Procrustes analysis involves finding the optimal superposition of two or more "forms" via rotations, translations, and scalings. Procrustes problems arise in a wide range of scientific disciplines, especially when the geometrical shapes of objects are compared, contrasted, and analyzed. Classically, the optimal transformations are found by minimizing the sum of the squared distances between corresponding points in the forms. Despite its widespread use, the ordinary unweighted least-squares (LS) criterion can give erroneous solutions when the errors have heterogeneous variances (heteroscedasticity) or the errors are correlated, both common occurrences with real data. In contrast, maximum likelihood (ML) estimation can provide accurate and consistent statistical estimates in the presence of both heteroscedasticity and correlation. Here we provide a complete solution to the nonisotropic ML Procrustes problem assuming a matrix Gaussian distribution with factored covariances. Our analysis generalizes, simplifies, and extends results from previous discussions of the ML Procrustes problem. An iterative algorithm is presented for the simultaneous, numerical determination of the ML solutions. PMID- 17130459 TI - Evidence for structural constraint on ovulin, a rapidly evolving Drosophila melanogaster seminal protein. AB - The egg-laying hormone ovulin (Acp26Aa) is among the most rapidly evolving proteins in the Drosophila genome. Against the background of ovulin's high sequence variability within and between species, we have identified highly conserved motifs that may play an important structural role. Using yeast two hybrid and GST-pull-down assays, we show that ovulin interacts with itself. The C terminus of ovulin is necessary and sufficient for self-interaction, with its C terminal 45 aa playing a major role. Under nonreducing conditions, ovulin participates in a high-molecular-mass complex, suggesting that it occurs in an oligomeric form. One or more of three predicted coiled-coil domains in the C terminus of ovulin may be involved in its self-interaction. These structural elements are conserved between species despite an overall rapid pace of evolution in ovulin's primary sequence. We therefore suggest that domains involved in ovulin's self-interaction form a conserved structural backbone for the protein, resulting in greater evolutionary flexibility at other sites. PMID- 17130460 TI - Time is brain. PMID- 17130462 TI - Comparative analysis of insulin gene promoters: implications for diabetes research. AB - DNA sequences that regulate expression of the insulin gene are located within a region spanning approximately 400 bp that flank the transcription start site. This region, the insulin promoter, contains a number of cis-acting elements that bind transcription factors, some of which are expressed only in the beta-cell and a few other endocrine or neural cell types, while others have a widespread tissue distribution. The sequencing of the genome of a number of species has allowed us to examine the manner in which the insulin promoter has evolved over a 450 million-year period. The major findings are that the A-box sites that bind PDX-1 are among the most highly conserved regulatory sequences, and that the conservation of the C1, E1, and CRE sequences emphasize the importance of MafA, E47/beta2, and cAMP-associated regulation. The review also reveals that of all the insulin gene promoters studied, the rodent insulin promoters are considerably dissimilar to the human, leading to the conclusion that extreme care should be taken when extrapolating rodent-based data on the insulin gene to humans. PMID- 17130463 TI - Probe-independent and direct quantification of insulin mRNA and growth hormone mRNA in enriched cell preparations. AB - Task division in multicellular organisms ensures that differentiated cell types produce cell-specific proteins that fulfill tasks for the whole organism. In some cases, the encoded mRNA species is so abundant that it represents a sizeable fraction of total mRNA in the cell. In this study, we have used a probe- and primer-free technique to quantify such abundant mRNA species in order to assess regulatory effects of in vitro and in vivo conditions. As a first example, we were able to quantify the regulation of proinsulin mRNA abundance in beta-cells by food intake or by the glucose concentration in tissue culture. The second example of application of this technique is the effect of corticosteroids on growth hormone mRNA in enriched somatrotrophs. It is anticipated that other examples exist in which measurement of very abundant mRNAs in dedicated cells will help to understand biological processes, monitor disease states, or assist biotechnological manufacturing procedures. PMID- 17130464 TI - Insulin-mediated phosphorylation of the proline-rich Akt substrate PRAS40 is impaired in insulin target tissues of high-fat diet-fed rats. AB - Clinical insulin resistance is associated with decreased activation of phosphatidylinositol 3'-kinase (PI3K) and its downstream substrate protein kinase B (PKB)/Akt. However, its physiological protein substrates remain poorly characterized. In the present study, the effect of in vivo insulin action on phosphorylation of the PKB/Akt substrate 40 (PRAS40) was examined. In rat and mice, insulin stimulated PRAS40-Thr246 phosphorylation in skeletal and cardiac muscle, the liver, and adipose tissue in vivo. Physiological hyperinsulinemia increased PRAS40-Thr246 phosphorylation in human skeletal muscle biopsies. In cultured cell lines, insulin-mediated PRAS40 phosphorylation was prevented by the PI3K inhibitors wortmannin and LY294002. Immunohistochemical and immunofluorescence studies showed that phosphorylated PRAS40 is predominantly localized to the nucleus. Finally, in rats fed a high-fat diet (HFD), phosphorylation of PRAS40 was markedly reduced compared with low-fat diet-fed animals in all tissues examined. In conclusion, the current study identifies PRAS40 as a physiological target of in vivo insulin action. Phosphorylation of PRAS40 is increased by insulin in human, rat, and mouse insulin target tissues. In rats, this response is reduced under conditions of HFD-induced insulin resistance. PMID- 17130465 TI - Fatty acid transport protein 1 is required for nonshivering thermogenesis in brown adipose tissue. AB - Nonshivering thermogenesis in brown adipose tissue (BAT) generates heat through the uncoupling of mitochondrial beta-oxidation from ATP production. The principal energy source for this process is fatty acids that are either synthesized de novo in BAT or are imported from circulation. How uptake of fatty acids is mediated and regulated has remained unclear. Here, we show that fatty acid transport protein (FATP)1 is expressed on the plasma membrane of BAT and is upregulated in response to cold stimuli, concomitant with an increase in the rate of fatty acid uptake. In FATP1-null animals, basal fatty acid uptake is reduced and remains unchanged following cold exposure. As a consequence, FATP1 knockout (KO) animals display smaller lipid droplets in BAT and fail to defend their core body temperature at 4 degrees C, despite elevated serum free fatty acid levels. Similarly, FATP1 is expressed by the BAT-derived cell line HIB-1B upon differentiation, and both fatty acid uptake and FATP1 protein levels are rapidly elevated following isoproterenol stimulation. Stimulation of fatty uptake by isoproterenol required both protein kinase A and mitogen-activated kinase signaling and is completely dependent on FATP1 expression, as small-hairpin RNA mediated knock down of FATP1 abrogated the effect. PMID- 17130466 TI - Effects of autoimmunity and immune therapy on beta-cell turnover in type 1 diabetes. AB - beta-Cell mass can expand in response to demand: during pregnancy, in the setting of insulin resistance, or after pancreatectomy. It is not known whether similar beta-cell hyperplasia occurs following immune therapy of autoimmune diabetes, but the clinical remission soon after diagnosis and the results of recent immune therapy studies suggest that beta-cell recovery is possible. We studied changes in beta-cell replication, mass, and apoptosis in NOD mice during progression to overt diabetes and following immune therapy with anti-CD3 monoclonal antibodies (mAbs) or immune regulatory T-cells (Tregs). beta-Cell replication increases in pre-diabetic mice, after adoptive transfer of diabetes with increasing islet inflammation but before an increase in blood glucose concentration or a significant decrease in beta-cell mass. The pathogenic cells are responsible for increasing beta-cell replication because replication was reduced during diabetes remission induced by anti-CD3 mAb or Tregs. beta-Cell replication stimulated by the initial inflammatory infiltrate results in increased production of new beta cells after immune therapy and increased beta-cell area, but the majority of this increased beta-cell area represents regranulated beta-cells rather than newly produced cells. We conclude that beta-cell replication is closely linked to the islet inflammatory process. A significant proportion of degranulated beta-cells remain, at the time of diagnosis of diabetes, that can recover after metabolic correction of hyperglycemia. Correction of the beta-cell loss in type 1 diabetes will, therefore, require strategies that target both the immunologic and cellular mechanisms that destroy and maintain beta-cell mass. PMID- 17130467 TI - Expression of IGF-I in pancreatic islets prevents lymphocytic infiltration and protects mice from type 1 diabetes. AB - Type 1 diabetic patients are diagnosed when beta-cell destruction is almost complete. Reversal of type 1 diabetes will require beta-cell regeneration from islet cell precursors and prevention of recurring autoimmunity. IGF-I expression in beta-cells of streptozotocin (STZ)-treated transgenic mice regenerates the endocrine pancreas by increasing beta-cell replication and neogenesis. Here, we examined whether IGF-I also protects islets from autoimmune destruction. Expression of interferon (IFN)-beta in beta-cells of transgenic mice led to islet beta(2)-microglobulin and Fas hyperexpression and increased lymphocytic infiltration. Pancreatic islets showed high insulitis, and these mice developed overt diabetes when treated with very-low doses of STZ, which did not affect control mice. IGF-I expression in IFN-beta-expressing beta-cells of double transgenic mice reduced beta(2)-microglobulin, blocked Fas expression, and counteracted islet infiltration. This was parallel to a decrease in beta-cell death by apoptosis in islets of STZ-treated IGF-I+IFN-beta-expressing mice. These mice were normoglycemic, normoinsulinemic, and showed normal glucose tolerance. They also presented similar pancreatic insulin content and beta-cell mass to healthy mice. Thus, local expression of IGF-I prevented islet infiltration and beta-cell death in mice with increased susceptibility to diabetes. These results indicate that pancreatic expression of IGF-I may regenerate and protect beta-cell mass in type 1 diabetes. PMID- 17130468 TI - Recovery of islet beta-cell function in streptozotocin- induced diabetic mice: an indirect role for the spleen. AB - Limitations in islet beta-cell transplantation as a therapeutic option for type 1 diabetes have prompted renewed interest in islet regeneration as a source of new islets. In this study we tested whether severely diabetic adult C57BL/6 mice can regenerate beta-cells. Diabetes was induced in C57BL/6 mice with high-dose streptozotocin (160-170 mg/kg). In the absence of islet transplantation, all diabetic mice remained diabetic (blood glucose >400 mg/dl), and no spontaneous reversal of diabetes was observed. When syngeneic islets (200/mouse) were transplanted into these diabetic mice under a single kidney capsule, stable restoration of euglycemia for >/=120 days was achieved. Removal of the kidney bearing the transplanted islets at 120 days posttransplantation revealed significant restoration of endogenous beta-cell function. This restoration of islet function was associated with increased beta-cell mass, as well as beta-cell hypertrophy and proliferation. The restoration of islet cell function was facilitated by the presence of a spleen; however, the facilitation was not due to the direct differentiation of spleen-derived cells into beta-cells. This study supports the possibility of restoring beta-cell function in diabetic individuals and points to a role for the spleen in facilitating this process. PMID- 17130469 TI - Maintenance of hepatic nuclear factor 6 in postnatal islets impairs terminal differentiation and function of beta-cells. AB - The Onecut homeodomain transcription factor hepatic nuclear factor 6 (Hnf6) is necessary for proper development of islet beta-cells. Hnf6 is initially expressed throughout the pancreatic epithelium but is downregulated in endocrine cells at late gestation and is not expressed in postnatal islets. Transgenic mice in which Hnf6 expression is maintained in postnatal islets (pdx1(PB)Hnf6) show overt diabetes and impaired glucose-stimulated insulin secretion (GSIS) at weaning. We now define the mechanism whereby maintenance of Hnf6 expression postnatally leads to beta-cell dysfunction. We provide evidence that continued expression of Hnf6 impairs GSIS by altering insulin granule biosynthesis, resulting in a reduced response to secretagogues. Sustained expression of Hnf6 also results in downregulation of the beta-cell-specific transcription factor MafA and a decrease in total pancreatic insulin. These results suggest that downregulation of Hnf6 expression in beta-cells during development is essential to achieve a mature, glucose-responsive beta-cell. PMID- 17130470 TI - The cell cycle inhibitory protein p21cip is not essential for maintaining beta cell cycle arrest or beta-cell function in vivo. AB - p21(cip1), a regulatory molecule upstream of the G(1/0) checkpoint, is increased in beta-cells in response to mitogenic stimulation. Whereas p21(cip1) can variably stimulate or inhibit cell cycle progression, in vitro studies suggest that p21(cip1) acts as an inhibitor in the pancreatic beta-cell. To determine the functional role of p21(cip1) in vivo, we studied p21-null mice. Surprisingly, islet mass, beta-cell replication rates, and function were normal in p21-null mice. We next attempted to drive beta-cell replication in p21-null mice by crossing them with rat insulin II promoter-murine PL-1 (islet-targeted placental lactogen transgenic) mice. Even with this added replicative stimulus of PL, p21 null islets showed no additional stimulation. A G(1/S) proteome scan demonstrated that p21(cip1) loss was not associated with compensatory increases in other cell cycle inhibitors (pRb, p107, p130, p16, p19, and p27), although mild increases in p57 were apparent. Surprisingly, p18, which had been anticipated to increase, was markedly decreased. In summary, isolated p21(cip1) loss, as for pRb, p53, p18, and p27 and other inhibitors, results in normal beta-cell development and function, either because it is not essential or because its function is subserved or complimented by another protein. These studies underscore marked inhibitory pressure and the complexity and plasticity of inhibitory pathways that restrain beta-cell replication. PMID- 17130471 TI - Protein inhibitor of neuronal nitric oxide synthase (PIN) is a new regulator of glucose-induced insulin secretion. AB - We previously showed that pancreatic beta-cells express neuronal nitric oxide synthase (nNOS) that controls insulin secretion through two catalytic activities: nitric oxide (NO) production and cytochrome c reductase activity. We now provide evidence that the endogenous protein inhibitor of nNOS (PIN) is expressed in rat pancreatic islets and INS-1 cells. Double-immunofluorescence studies showed a colocalization of PIN with both nNOS and myosin Va in insulin-secreting beta cells. Electron microscopy studies confirmed that PIN is mainly associated with insulin secretory granules and colocated with nNOS in the latter. In addition, PIN overexpression in INS-1 cells enhanced glucose-induced insulin secretion, which is only partly reversed by addition of an NO donor, sodium nitroprusside (SNP), and unaffected by the inhibitor of cytochrome c reductase activity, miconazole. In contrast, the pharmacological inhibitor of nNOS, Nomega-nitro-l arginine methyl ester, amplified glucose-induced insulin secretion, an effect insensitive to SNP but completely normalized by the addition of miconazole. Thus, PIN insulinotropic effect could be related to its colocalization with the actin based molecular motor myosin Va and as such be implicated in the physiological regulation of glucose-induced insulin secretion at the level of the exocytotic machinery. PMID- 17130472 TI - Regulation of pancreatic beta-cell regeneration in the normoglycemic 60% partial pancreatectomy mouse. AB - beta-Cell mass is determined by a dynamic balance of proliferation, neogenesis, and apoptosis. The precise mechanisms underlying compensatory beta-cell mass (BCM) homeostasis are not fully understood. To evaluate the processes that maintain normoglycemia and regulate BCM during pancreatic regeneration, C57BL/6 mice were analyzed for 15 days following 60% partial pancreatectomy (Px). BCM increased in Px mice from 2 days onwards and was approximately 68% of the shams by 15 days, partly due to enhanced beta-cell proliferation. A transient approximately 2.8-fold increase in the prevalence of beta-cell clusters/small islets at 2 days post-Px contributed substantially to BCM augmentation, followed by an increase in the number of larger islets at 15 days. To evaluate the signaling mechanisms that may regulate this compensatory growth, we examined key intermediates of the insulin signaling pathway. We found insulin receptor substrate (IRS)2 and enhanced-activated Akt immunoreactivity in islets and ducts that correlated with increased pancreatic duodenal homeobox (PDX)1 expression. In contrast, forkhead box O1 expression was decreased in islets but increased in ducts, suggesting distinct PDX1 regulatory mechanisms in these tissues. Px animals acutely administered insulin exhibited further enhancement in insulin signaling activity. These data suggest that the IRS2-Akt pathway mediates compensatory beta-cell growth by activating beta-cell proliferation with an increase in the number of beta-cell clusters/small islets. PMID- 17130473 TI - Downregulation of EGF receptor signaling in pancreatic islets causes diabetes due to impaired postnatal beta-cell growth. AB - Epidermal growth factor receptor (EGF-R) signaling is essential for proper fetal development and growth of pancreatic islets, and there is also evidence for its involvement in beta-cell signal transduction in the adult. To study the functional roles of EGF-R in beta-cell physiology in postnatal life, we have generated transgenic mice that carry a mutated EGF-R under the pancreatic duodenal homeobox-1 promoter (E1-DN mice). The transgene was expressed in islet beta- and delta-cells but not in alpha-cells, as expected, and it resulted in an approximately 40% reduction in pancreatic EGF-R, extracellular signal-related kinase, and Akt phosphorylation. Homozygous E1-DN mice were overtly diabetic after the age of 2 weeks. The hyperglycemia was more pronounced in male than in female mice. The relative beta-cell surface area of E1-DN mice was highly reduced at the age of 2 months, while alpha-cell surface area was not changed. This defect was essentially postnatal, since the differences in beta-cell area of newborn mice were much smaller. An apparent explanation for this is impaired postnatal beta-cell proliferation; the normal surge of beta-cell proliferation during 2 weeks after birth was totally abolished in the transgenic mice. Heterozygous E1-DN mice were glucose intolerant in intraperitoneal glucose tests. This was associated with a reduced insulin response. However, downregulation of EGF-R signaling had no influence on the insulinotropic effect of glucagon-like peptide-1 analog exendin-4. In summary, our results show that even a modest attenuation of EGF-R signaling leads to a severe defect in postnatal growth of the beta-cells, which leads to the development of diabetes. PMID- 17130474 TI - Skeletal muscle mitochondrial functions, mitochondrial DNA copy numbers, and gene transcript profiles in type 2 diabetic and nondiabetic subjects at equal levels of low or high insulin and euglycemia. AB - We investigated whether previously reported muscle mitochondrial dysfunction and altered gene transcript levels in type 2 diabetes might be secondary to abnormal blood glucose and insulin levels rather than an intrinsic defect of type 2 diabetes. A total of 13 type 2 diabetic and 17 nondiabetic subjects were studied on two separate occasions while maintaining similar insulin and glucose levels in both groups by 7-h infusions of somatostatin, low- or high-dose insulin (0.25 and 1.5 mU/kg of fat-free mass per min, respectively), and glucose. Muscle mitochondrial DNA abundance was not different between type 2 diabetic and nondiabetic subjects at both insulin levels, but the majority of transcripts in muscle that are involved mitochondrial functions were expressed at lower levels in type 2 diabetes at low levels of insulin. However, several gene transcripts that are specifically involved in the electron transport chain were expressed at higher levels in type 2 diabetic patients. After the low-dose insulin infusion, which achieved postabsorptive insulin levels, the muscle mitochondrial ATP production rate (MAPR) was not different between type 2 diabetic and nondiabetic subjects. However, increasing insulin to postprandial levels increased the MAPR in nondiabetic subjects but not in type 2 diabetic patients. The lack of MAPR increment in response to high-dose insulin in type 2 diabetic patients occurred in association with reduced glucose disposal and expression of peroxisome proliferator-activated receptor-gamma coactivator 1alpha, citrate synthase, and cytochrome c oxidase I. In conclusion, the current data supports that muscle mitochondrial dysfunction in type 2 diabetes is not an intrinsic defect, but instead a functional defect related to impaired response to insulin. PMID- 17130475 TI - Tau is hyperphosphorylated at multiple sites in mouse brain in vivo after streptozotocin-induced insulin deficiency. AB - Deficient signaling by insulin, as occurs in diabetes, is associated with impaired brain function, and diabetes is associated with an increased prevalence of Alzheimer's disease. One of the hallmark pathological characteristics of Alzheimer's disease is the presence of neurofibrillary tangles containing hyperphosphorylated tau, a microtubule-associated protein. Therefore, we tested the hypothesis that insulin depletion caused by administration of streptozotocin may cause tau hyperphosphorylation in mouse brain by using site-specific phosphorylation-dependent tau antibodies to obtain precise identification of the phosphorylation of tau on individual residues. A massive (fivefold average increase) and widespread at multiple residues (detected with eight different phosphorylation-dependent tau antibodies) increase in the phosphorylation of tau was found in mouse cerebral cortex and hippocampus within 3 days of insulin depletion by streptozotocin treatment. This hyperphosphorylation of tau at some sites was rapidly reversible by peripheral insulin administration. Examination of several kinases that phosphorylate tau indicated that they were unlikely to account for the widespread hyperphosphorylation of tau caused by streptozotocin treatment, but there was a large decrease in mouse brain protein phosphatase 2A activity, which is known to mediate tau phosphorylation. These results show that insulin deficiency causes rapid and large increases in tau phosphorylation, a condition that could prime tau for the neuropathology of Alzheimer's disease, thereby contributing to the increased susceptibility to Alzheimer's disease caused by diabetes. PMID- 17130476 TI - Hyperglycemia inhibits retinoic acid-induced activation of Rac1, prevents differentiation of cortical neurons, and causes oxidative stress in a rat model of diabetic pregnancy. AB - Diabetes is a risk factor for neuronal dysfunction. Impairment in signaling mechanisms that regulate differentiation of neurons is hypothesized to be one of the main causes of neuronal dysfunction. Retinoic acid, a physiologically active retinoid synthesized from vitamin A, regulates neuronal differentiation during embryonic development and is required for maintenance of plasticity in differentiated neurons. To date, little is known about the molecular events underlying hyperglycemia-induced complications in the central nervous system (CNS). Here, we provide evidence, in a diabetes rat model, of hyperglycemia induced oxidative stress along with apoptotic stress in developing cortical neurons isolated from 16-day-old rat embryos. We also demonstrate impaired retinoic acid signaling that is involved in neuronal differentiation. Retinoic acid-induced neurite outgrowth and expression of neuronal markers were reduced in this model. The activation of small-molecular weight G-protein, Rac1, that mediates these effects was also reduced. Retinoic acid applied at a physiological concentration significantly decreased hyperglycemia-induced oxidative stress and thus supported the antioxidant defense system. These results suggest that diabetes-induced neuronal complications during pregnancy might be due to impaired retinoic acid signaling, and exogenously administered retinoic acid may be useful against CNS complications associated with diabetes. PMID- 17130477 TI - The leptin-deficient (ob/ob) mouse: a new animal model of peripheral neuropathy of type 2 diabetes and obesity. AB - Whereas functional, metabolic, neurotrophic, and morphological abnormalities of peripheral diabetic neuropathy (PDN) have been extensively explored in streptozotocin-induced diabetic rats and mice (models of type 1 diabetes), insufficient information is available on manifestations and pathogenetic mechanisms of PDN in type 2 diabetic models. The latter could constitute a problem for clinical trial design because the vast majority of subjects with diabetes have type 2 (non-insulin dependent) diabetes. This study was aimed at characterization of PDN in leptin-deficient (ob/ob) mice, a model of type 2 diabetes with relatively mild hyperglycemia and obesity. ob/ob mice ( approximately 11 weeks old) clearly developed manifest sciatic motor nerve conduction velocity (MNCV) and hind-limb digital sensory nerve conduction velocity (SNCV) deficits, thermal hypoalgesia, tactile allodynia, and a remarkable ( approximately 78%) loss of intraepidermal nerve fibers. They also had increased sorbitol pathway activity in the sciatic nerve and increased nitrotyrosine and poly(ADP-ribose) immunofluorescence in the sciatic nerve, spinal cord, and dorsal root ganglion (DRG). Aldose reductase inhibition with fidarestat (16 mg . kg(-1) . d(-1)), administered to ob/ob mice for 6 weeks starting from 5 weeks of age, was associated with preservation of normal MNCV and SNCV and alleviation of thermal hypoalgesia and intraepidermal nerve fiber loss but not tactile allodynia. Sciatic nerve nitrotyrosine immunofluorescence and the number of poly(ADP-ribose)-positive nuclei in sciatic nerve, spinal cord, and DRGs of fidarestat-treated ob/ob mice did not differ from those in nondiabetic controls. In conclusion, the leptin-deficient ob/ob mouse is a new animal model that develops both large motor and sensory fiber and small sensory fiber PDN and responds to pathogenetic treatment. The results support the role for increased aldose reductase activity in functional and structural changes of PDN in type 2 diabetes. PMID- 17130478 TI - Feasibility of automating insulin delivery for the treatment of type 1 diabetes. AB - An automated closed-loop insulin delivery system based on subcutaneous glucose sensing and subcutaneous insulin delivery was evaluated in 10 subjects with type 1 diabetes (2 men, 8 women, mean [+/-SD] age 43.4 +/- 11.4 years, duration of diabetes 18.2 +/- 13.5 years). Closed-loop control was assessed over approximately 30 h and compared with open-loop control assessed over 3 days. Closed-loop insulin delivery was calculated using a model of the beta-cell's multiphasic insulin response to glucose. Plasma glucose was 160 +/- 66 mg/dl at the start of closed loop and was thereafter reduced to 71 +/- 19 by 1:00 p.m. (preprandial lunch). Fasting glucose the subsequent morning on closed loop was not different from target (124 +/- 25 vs. 120 mg/dl, respectively; P > 0.05). Mean glucose levels were not different between the open and closed loop (133 +/- 63 vs. 133 +/- 52 mg/dl, respectively; P > 0.65). However, glucose was within the range 70-180 mg/dl 75% of the time under closed loop versus 63% for open loop. Incidence of biochemical hypoglycemia (blood glucose <60 mg/dl) was similar under the two treatments. There were no episodes of severe hypoglycemia. The data provide proof of concept that glycemic control can be achieved by a completely automated external closed-loop insulin delivery system. PMID- 17130479 TI - Introgression of F344 rat genomic DNA on BB rat chromosome 4 generates diabetes resistant lymphopenic BB rats. AB - Failure to express the Gimap5 protein is associated with lymphopenia (lyp) and linked to spontaneous diabetes in the diabetes-prone BioBreeding (BBDP) rat. Gimap5 is a member of seven related genes located within 150 Kb on rat chromosome 4. Congenic DR.(lyp/lyp) rats, where BBDP lyp was introgressed onto the diabetes resistant BBDR background (BBDR.BBDP.(lyp/lyp)), all develop diabetes between 46 and 81 days of age (mean +/- SE, 61 +/- 1), whereas DR.(lyp/+) and DR.(+/+) rats are nonlymphopenic and diabetes resistant. In an intercross between F1(BBDP x F344) rats, we identified a rat with a recombination event on chromosome 4, allowing us to fix 33 Mb of F344 between D4Rat253 and D4Rhw6 in the congenic DR.lyp rat line. Gimap1 and Gimap5 were the only members of the Gimap family remaining homozygous for the BBDP allele. Offspring homozygous for the F344 allele (f/f) between D4Rat253 and D4Rhw6 were lymphopenic (85 of 85, 100%) but did not develop diabetes (0 of 85). During rescue of the recombination, 102 of 163 (63%) rats heterozygous (b/f) for the recombination developed diabetes between 52 and 222 days of age (88 +/- 3). Our data demonstrate that introgression of a 33-Mb region of the F344 genome, proximal to the mutated Gimap5 gene, renders the rat diabetes resistant despite being lymphopenic. Spontaneous diabetes in the BB rat may therefore be controlled, in part, by a diabetogenic factor(s), perhaps unrelated to the Gimap5 mutation on rat chromosome 4. PMID- 17130480 TI - A genome-wide linkage scan for genes controlling variation in renal function estimated by serum cystatin C levels in extended families with type 2 diabetes. AB - We performed a variance components linkage analysis of renal function, measured as glomerular filtration rate (GFR), in 63 extended families with multiple members with type 2 diabetes. GFR was estimated from serum concentrations of cystatin C and creatinine in 406 diabetic and 428 nondiabetic relatives. Results for cystatin C were summarized because they are superior to creatinine results. GFR aggregates in families with significant heritability (h(2)) in diabetic (h(2) = 0.45, P < 1 x 10(-5)) and nondiabetic (h(2) = 0.36, P < 1 x 10(-3)) relatives. Genetic correlation (r(G) = 0.35) between the GFR of diabetic and nondiabetic relatives was less than one (P = 0.01), suggesting that genes controlling GFR variation in these groups are different. Linkage results supported this interpretation. In diabetic relatives, linkage was strong on chromosome 2q (logarithm of odds [LOD] = 4.1) and suggestive on 10q (LOD = 3.1) and 18p (LOD = 2.2). In nondiabetic relatives, linkage was suggestive on 3q (LOD = 2.2) and 11p (LOD = 2.1). When diabetic and nondiabetic relatives were combined, strong evidence for linkage was found only on 7p (LOD = 4.0). In conclusion, partially distinct sets of genes control GFR variation in relatives with and without diabetes on chromosome 2q, possibly on 10q and 18p in the former, and on 7p in both. None of these genes overlaps with genes controlling variation in urinary albumin excretion. PMID- 17130481 TI - Hyperphagia, severe obesity, impaired cognitive function, and hyperactivity associated with functional loss of one copy of the brain-derived neurotrophic factor (BDNF) gene. AB - The neurotrophin brain-derived neurotrophic factor (BDNF) inhibits food intake, and rodent models of BDNF disruption all exhibit increased food intake and obesity, as well as hyperactivity. We report an 8-year-old girl with hyperphagia and severe obesity, impaired cognitive function, and hyperactivity who harbored a de novo chromosomal inversion, 46,XX,inv(11)(p13p15.3), a region encompassing the BDNF gene. We have identified the proximal inversion breakpoint that lies 850 kb telomeric of the 5' end of the BDNF gene. The patient's genomic DNA was heterozygous for a common coding polymorphism in BDNF, but monoallelic expression was seen in peripheral lymphocytes. Serum concentration of BDNF protein was reduced compared with age- and BMI-matched subjects. Haploinsufficiency for BDNF was associated with increased ad libitum food intake, severe early-onset obesity, hyperactivity, and cognitive impairment. These findings provide direct evidence for the role of the neurotrophin BDNF in human energy homeostasis, as well as in cognitive function, memory, and behavior. PMID- 17130482 TI - Decreased hepatic futile cycling compensates for increased glucose disposal in the Pten heterodeficient mouse. AB - Despite altered regulation of insulin signaling, Pten(+/-) heterodeficient standard diet-fed mice, approximately 4 months old, exhibit normal fasting glucose and insulin levels. We report here a stable isotope flux phenotyping study of this "silent" phenotype, in which tissue-specific insulin effects in whole-body Pten(+/-)-deficient mice were dissected in vivo. Flux phenotyping showed gain of function in Pten(+/-) mice, seen as increased peripheral glucose disposal, and compensation by a metabolic feedback mechanism that 1) decreases hepatic glucose recycling via suppression of glucokinase expression in the basal state to preserve hepatic glucose production and 2) increases hepatic responsiveness in the fasted-to-fed transition. In Pten(+/-) mice, hepatic gene expression of glucokinase was 10-fold less than wild-type (Pten(+/+)) mice in the fasted state and reached Pten(+/+) values in the fed state. Glucose-6-phosphatase expression was the same for Pten(+/-) and Pten(+/+) mice in the fasted state, and its expression for Pten(+/-) was 25% of Pten(+/+) in the fed state. This study demonstrates how intra- and interorgan flux compensations can preserve glucose homeostasis (despite a specific gene defect that accelerates glucose disposal) and how flux phenotyping can dissect these tissue-specific flux compensations in mice presenting with a "silent" phenotype. PMID- 17130483 TI - Sodium-coupled glucose cotransporters contribute to hypothalamic glucose sensing. AB - Specialized neurons within the hypothalamus have the ability to sense and respond to changes in ambient glucose concentrations. We investigated the mechanisms underlying glucose-triggered activity in glucose-excited neurons, using primary cultures of rat hypothalamic neurons monitored by fluorescence calcium imaging. We found that 35% (738 of 2,139) of the neurons were excited by increasing glucose from 3 to 15 mmol/l, but only 9% (6 of 64) of these glucose-excited neurons were activated by tolbutamide, suggesting the involvement of a ATP sensitive K(+) channel-independent mechanism. alpha-Methylglucopyranoside (alphaMDG; 12 mmol/l), a nonmetabolizable substrate of sodium glucose cotransporters (SGLTs), mimicked the effect of high glucose in 67% of glucose excited neurons, and both glucose- and alphaMDG-triggered excitation were blocked by Na(+) removal or by the SGLT inhibitor phloridzin (100 nmol/l). In the presence of 0.5 mmol/l glucose and tolbutamide, responses could also be triggered by 3.5 mmol/l alphaMDG, supporting a role for an SGLT-associated mechanism at low as well as high substrate concentrations. Using RT-PCR, we detected SGLT1, SGLT3a, and SGLT3b in both cultured neurons and adult rat hypothalamus. Our findings suggest a novel role for SGLTs in glucose sensing by hypothalamic glucose-excited neurons. PMID- 17130484 TI - Leptin regulation of the anorexic response to glucagon-like peptide-1 receptor stimulation. AB - Leptin reduces food intake in part by enhancing satiety responses to gastrointestinal signals produced in response to food consumption. Glucagon-like peptide 1 (GLP-1), secreted by the intestine when nutrients enter the gut, is one such putative satiety signal. To investigate whether leptin enhances the anorexic effects of GLP-1, rats received either saline or a subthreshold dose of leptin before intraperitoneal injection of either GLP-1 or Exendin-4 (Ex4; a GLP-1 receptor agonist). Leptin pretreatment strongly enhanced anorexia and weight loss induced by GLP-1 or Ex4 over 24 h. Conversely, fasting attenuated the anorexic response to GLP-1 or Ex4 treatment via a leptin-dependent mechanism, as demonstrated by our finding that the effect of fasting was reversed by physiological leptin replacement. As expected, Ex4 induced expression of c-Fos protein, a marker of neuronal activation, in hindbrain areas that process afferent input from satiety signals, including the nucleus of the solitary tract and area postrema. Unexpectedly, leptin pretreatment blocked this response. These findings identify physiological variation of plasma leptin levels as a potent regulator of GLP-1 receptor-mediated food intake suppression and suggest that the underlying mechanism is distinct from that which mediates interactions between leptin and other satiety signals. PMID- 17130485 TI - Endogenous ApoE expression modulates adipocyte triglyceride content and turnover. AB - Apolipoprotein E (apoE) is highly expressed in adipose tissue and adipocytes in which its expression is regulated by peroxisome proliferator-activated receptor (PPAR)-gamma agonists and tumor necrosis factor-alpha. There is, however, no information regarding a role for endogenous apoE in differentiated adipocyte function. In this report, we define a novel role for apoE in modulating adipocyte lipid metabolism. ApoE(-/-) mice have less body fat and smaller adipocytes compared with wild-type controls. Freshly isolated adipose tissue from apoE(-/-) mice contains lower levels of triglyceride and free fatty acid, and these differences are maintained in cultured adipocytes derived from preadipocytes. Adenoviral expression of apoE in apoE(-/-)-cultured adipocytes increases triglyceride and fatty acid content. During incubation with apoE-containing triglyceride-rich lipoproteins, apoE(-/-) adipose tissue accumulates less triglyceride than wild type. The absence of apoE expression in primary cultured adipocytes also leads to changes in the expression of genes involved in the metabolism/turnover of fatty acids and the triglyceride droplet. Markers of adipocyte differentiation were lower in freshly isolated and cultured apoE(-/-) adipocytes. Importantly, PPAR-gamma-mediated changes in lipid content and gene expression are markedly altered in cultured apoE(-/-) adipocytes. These results establish a novel role for endogenous apoE in adipocyte lipid metabolism and have implications for constructing an integrated model of adipocyte physiology in health and disease. PMID- 17130486 TI - Effects of rimonabant (SR141716) on fasting-induced hypothalamic-pituitary adrenal axis and neuronal activation in lean and obese Zucker rats. AB - The effects of the cannabinoid-1 receptor (CB(1)) antagonist rimonabant on energy metabolism and fasting-induced hypothalamic-pituitary-adrenal (HPA) axis and neuronal activation were investigated. Lean and obese Zucker rats were treated orally with a daily dose of 10 mg/kg rimonabant for 14 days. A comprehensive energy balance profile based on whole-carcass analyses further demonstrated the potential of CB(1) antagonists for decreasing energy gain through reducing food intake and potentially increasing brown adipose tissue thermogenesis. Rimonabant also reduced plasma glucose, insulin, and homeostasis model assessment of insulin resistance, which further confirms the ability of CB(1) antagonists to improve insulin sensitivity. To test the hypothesis that rimonabant attenuates the effect of fasting on HPA axis activation in the obese Zucker model, rats were either ad libitum-fed or food-deprived for 8 h. Contrary to expectation, rimonabant increased basal circulating corticosterone levels and enhanced the HPA axis response to food deprivation in obese rats. Rimonabant also exacerbated the neuronal activation seen in the arcuate nucleus (ARC) after short-term deprivation. In conclusion, the present study demonstrates that CB(1) blockade does not prevent the hypersensitivity to food deprivation occurring at the level of HPA axis and ARC activation in the obese Zucker rats. This, however, does not prevent CB(1) antagonism from exerting beneficial effects on energy and glucose metabolism. PMID- 17130487 TI - Genetic ablation of the c-Cbl ubiquitin ligase domain results in increased energy expenditure and improved insulin action. AB - Casitas b-lineage lymphoma (c-Cbl) is a multiadaptor protein with E3-ubiquitin ligase activity residing within its RING finger domain. We have previously reported that c-Cbl-deficient mice exhibit elevated energy expenditure, reduced adiposity, and improved insulin action. In this study, we examined mice expressing c-Cbl protein with a loss-of-function mutation within the RING finger domain (c-Cbl(A/-) mice). Compared with control animals, c-Cbl(A/-) mice display a phenotype that includes reduced adiposity, despite greater food intake; reduced circulating insulin, leptin, and triglyceride levels; and improved glucose tolerance. c-Cbl(A/-) mice also display elevated oxygen consumption (13%) and are protected against high-fat diet-induced obesity and insulin resistance. Unlike c Cbl(A/-) mice, mice expressing a mutant c-Cbl with the phosphatidylinositol (PI) 3-kinase binding domain ablated (c-Cbl(F/F) mice) exhibited an insulin sensitivity, body composition, and energy expenditure similar to that of wild type animals. These results indicate that c-Cbl ubiquitin ligase activity, but not c-Cbl-dependent activation of PI 3-kinase, plays a key role in the regulation of whole-body energy metabolism. PMID- 17130488 TI - OXPAT/PAT-1 is a PPAR-induced lipid droplet protein that promotes fatty acid utilization. AB - Lipid droplet proteins of the PAT (perilipin, adipophilin, and TIP47) family regulate cellular neutral lipid stores. We have studied a new member of this family, PAT-1, and found that it is expressed in highly oxidative tissues. We refer to this protein as "OXPAT." Physiologic lipid loading of mouse liver by fasting enriches OXPAT in the lipid droplet tissue fraction. OXPAT resides on lipid droplets with the PAT protein adipophilin in primary cardiomyocytes. Ectopic expression of OXPAT promotes fatty acid-induced triacylglycerol accumulation, long-chain fatty acid oxidation, and mRNAs associated with oxidative metabolism. Consistent with these observations, OXPAT is induced in mouse adipose tissue, striated muscle, and liver by physiological (fasting), pathophysiological (insulin deficiency), pharmacological (peroxisome proliferator activated receptor [PPAR] agonists), and genetic (muscle-specific PPARalpha overexpression) perturbations that increase fatty acid utilization. In humans with impaired glucose tolerance, PPARgamma agonist treatment induces adipose OXPAT mRNA. Further, adipose OXPAT mRNA negatively correlates with BMI in nondiabetic humans. Our collective data in cells, mice, and humans suggest that OXPAT is a marker for PPAR activation and fatty acid oxidation. OXPAT likely contributes to adaptive responses to the fatty acid burden that accompanies fasting, insulin deficiency, and overnutrition, responses that are defective in obesity and type 2 diabetes. PMID- 17130489 TI - Lipin deficiency impairs diurnal metabolic fuel switching. AB - Fatty liver is a common feature of both obesity and lipodystrophy, reflecting compromised adipose tissue function. The lipin-deficient fatty liver dystrophy (fld) mouse is an exception, as there is lipodystrophy without a fatty liver. Using a combination of indirect calorimetry and stable-isotope flux phenotyping, we determined that fld mice exhibit abnormal fuel utilization throughout the diurnal cycle, with increased glucose oxidation near the end of the fasting period and increased fatty acid oxidation during the feeding period. The mechanisms underlying these alterations include a twofold increase compared with wild-type mice in tissue glycogen storage during the fed state, a 40% reduction in hepatic glucose production in the fasted state, and a 27-fold increase in de novo fatty acid synthesis in liver during the fed state. Thus, the inability to store energy in adipose tissue in the fld mouse leads to a compensatory increase in glycogen storage for use during the fasting period and reliance upon hepatic fatty acid synthesis to provide fuel for peripheral tissues during the fed state. The increase in hepatic fatty acid synthesis and peripheral utilization provides a potential mechanism to ameliorate fatty liver in the fld that would otherwise occur as a consequence of adipose tissue dysfunction. PMID- 17130490 TI - Altered monocyte cyclooxygenase response to lipopolysaccharide in type 1 diabetes. AB - Type 1 diabetes is caused by adaptive immune responses, but innate immunity is important because monocytes infiltrate islets. Activated monocytes express cyclooxygenase (COX)-2, promoting prostaglandin-E(2) (PGE(2)) secretion, whereas COX-1 expression is constitutive. We aimed to define monocyte COX expression in type 1 diabetes basally and after lipopolysaccharide (LPS) stimulation. Isolated CD14(+) monocytes were analyzed for COX mRNA and protein expression from identical twins (discordant for type 1 diabetes) and control subjects. Basal monocyte COX mRNA, protein expression, and PGE(2) secretion were normal in type 1 diabetic subjects. After LPS, twins and control subjects showed a COX mRNA isoform switch with decreased COX-1 mRNA (P < 0.01), increased COX-2 mRNA (P < 0.01), and increased COX-2 protein expression (P < 0.01). Compared with control subjects, both diabetic and nondiabetic twins showed greater LPS-induced downregulation of monocyte COX-1 mRNA (P = 0.02), reduced upregulation of COX-2 mRNA and protein (P < 0.03), and greater inhibition by the COX-2 inhibitor di isopropylfluorophosphate (DFP) of monocyte PGE(2) (P < 0.007). We demonstrate an alteration in monocyte COX mRNA expression as well as monocyte COX-2 and PGE(2) production after LPS in type 1 diabetic patients and their nondiabetic twins. Because COX-2 response to LPS is proinflammatory, an inherited reduced response would predispose to chronic inflammatory diseases such as type 1 diabetes. PMID- 17130491 TI - Insulin treatment in patients with type 1 diabetes induces upregulation of regulatory T-cell markers in peripheral blood mononuclear cells stimulated with insulin in vitro. AB - Patients with type 1 diabetes are treated with daily injections of human insulin, an autoantigen expressed in thymus. Natural CD4(+)CD25(high) regulatory T-cells are derived from thymus, and accordingly human insulin-specific regulatory T cells should exist. We had a chance to study peripheral blood mononuclear cells (PBMCs) from children with type 1 diabetes both before and after starting insulin treatment, and thus we could analyze the effects of insulin treatment on regulatory T-cells in children with type 1 diabetes. PBMCs were stimulated for 72 h with bovine/human insulin. The mRNA expression of regulatory T-cell markers (transforming growth factor-beta, Foxp3, cytotoxic T-lymphocyte antigen-4 [CTLA 4], and inducible co-stimulator [ICOS]) or cytokines (gamma-interferon [IFN gamma], interleukin [IL]-5, IL-4) was measured by quantitative RT-PCR. The secretion of IFN-gamma, IL-2, IL-4, IL-5, and IL-10 was also studied. The expression of Foxp3, CTLA-4, and ICOS mRNAs in PBMCs stimulated with bovine or human insulin was higher in patients on insulin treatment than in patients studied before starting insulin treatment. The insulin-induced Foxp3 protein expression in CD4(+)CD25(high) cells was detectable in flow cytometry. No differences were seen in cytokine activation between the patient groups. Insulin stimulation in vitro induced increased expression of regulatory T-cell markers, Foxp3, CTLA-4, and ICOS only in patients treated with insulin, suggesting that treatment with human insulin activates insulin-specific regulatory T-cells in children with newly diagnosed type 1 diabetes. This effect of the exogenous autoantigen could explain the difficulties to detect in vitro T-cell proliferation responses to insulin in newly diagnosed patients. Furthermore, autoantigen treatment-induced activation of regulatory T-cells may contribute to the clinical remission of the disease. PMID- 17130492 TI - Modulatory role of DR4- to DQ8-restricted CD4 T-cell responses and type 1 diabetes susceptibility. AB - This study addressed an important biological question, namely how certain HLA molecules modulate the disease risk conferred by other HLA molecules. The HLA molecules under investigation were HLA-DQ8 and -DR4, the two most prevalent HLA class II alleles found in Caucasian type 1 diabetic patients. A panel of human GAD (hGAD65)-specific CD4 T-cell lines and hybridomas was generated to serve as detection reagents for evaluating the peptide occupancy of DQ8 and DR4. Results indicated that DQ8 and DR4 (0401) were able to bind the same hGAD65 peptides. The coexpression of DR4 (0401) diminished DQ8-restricted T-cell responses. In addition, we also demonstrated that the diminished T-cell response varied according to the specific DRB1*04 alleles. Taken together, this study provides evidence that DR4 is able to modulate DQ8-restricted T-cell responses, possibly by competing for peptides. Given that DQ8 is a primary genetic determinant of type 1 diabetes, the decreased DQ8-restricted CD4 T-cell activity due to peptide competition may be the mechanism explaining the modulation effect of DR4 to type 1 diabetes susceptibility. PMID- 17130493 TI - Glucagon receptor knockout mice display increased insulin sensitivity and impaired beta-cell function. AB - In previous studies, glucagon receptor knockout mice (Gcgr(-/-)) display reduced blood glucose and increased glucose tolerance, with hyperglucagonemia and increased levels of glucagon-like peptide (GLP)-1. However, the role of glucagon receptor signaling for the regulation of islet function and insulin sensitivity is unknown. We therefore explored beta-cell function and insulin sensitivity in Gcgr(-/-) and wild-type mice. The steady-state glucose infusion rate during hyperinsulinemic-euglycemic clamp was elevated in Gcgr(-/-) mice, indicating enhanced insulin sensitivity. Furthermore, the acute insulin response (AIR) to intravenous glucose was higher in Gcgr(-/-) mice. The augmented AIR to glucose was blunted by the GLP-1 receptor antagonist, exendin-3. In contrast, AIR to intravenous administration of other secretagogues was either not affected (carbachol) or significantly reduced (arginine, cholecystokinin octapeptide) in Gcgr(-/-) mice. In islets isolated from Gcgr(-/-) mice, the insulin responses to glucose and several insulin secretagogues were all significantly blunted compared with wild-type mice. Furthermore, glucose oxidation was reduced in islets from Gcgr(-/-) mice. In conclusion, the present study shows that glucagon signaling is required for normal beta-cell function and that insulin action is improved when disrupting the signal. In vivo, augmented GLP-1 levels compensate for the impaired beta-cell function in Gcgr(-/-) mice. PMID- 17130494 TI - Nutrient control of insulin secretion in isolated normal human islets. AB - Pancreatic islets were isolated from 16 nondiabetic organ donors and, after culture for approximately 2 days in 5 mmol/l glucose, were perifused to characterize nutrient-induced insulin secretion in human islets. Stepwise increases from 0 to 30 mmol/l glucose (eight 30-min steps) evoked concentration dependent insulin secretion with a threshold at 3-4 mmol/l glucose, K(m) at 6.5 mmol/l glucose, and V(max) at 15 mmol/l glucose. An increase from 1 to 15 mmol/l glucose induced biphasic insulin secretion with a prominent first phase (peak increase of approximately 18-fold) and a sustained, flat second phase ( approximately 10-fold increase), which were both potentiated by forskolin. The central role of ATP-sensitive K(+) channels in the response to glucose was established by abrogation of insulin secretion by diazoxide and reversible restoration by tolbutamide. Depolarization with tolbutamide or KCl (plus diazoxide) triggered rapid insulin secretion in 1 mmol/l glucose. Subsequent application of 15 mmol/l glucose further increased insulin secretion, showing that the amplifying pathway is operative. In control medium, glutamine alone was ineffective, but its combination with leucine or nonmetabolized 2-amino-bicyclo [2,2,1]-heptane-2-carboxylic acid (BCH) evoked rapid insulin secretion. The effect of BCH was larger in low glucose than in high glucose. In contrast, the insulin secretion response to arginine or a mixture of four amino acids was potentiated by glucose or tolbutamide. Palmitate slightly augmented insulin secretion only at the supraphysiological palmitate-to-albumin ratio of 5. Inosine and membrane-permeant analogs of pyruvate, glutamate, or succinate increased insulin secretion in 3 and 10 mmol/l glucose, whereas lactate and pyruvate had no effect. In conclusion, nutrient-induced insulin secretion in normal human islets is larger than often reported. Its characteristics are globally similar to those of insulin secretion by rodent islets, with both triggering and amplifying pathways. The pattern of the biphasic response to glucose is superimposable on that in mouse islets, but the concentration-response curve is shifted to the left, and various nutrients, in particular amino acids, influence insulin secretion within the physiological range of glucose concentrations. PMID- 17130495 TI - Modulation of insulin secretion by fatty acyl analogs. AB - The secretagogue, the incretin-like, and the suppressive activities of long-chain fatty acids (LCFAs) in modulating insulin secretion in vivo and in cultured islets were simulated here by beta,beta'-tetramethyl-hexadecanedioic acid (M16) and alpha,alpha'-tetrachloro-tetradecanedioic acid (Cl-DICA). M16, but not Cl DICA, serves as a substrate for ATP-dependent CoA thioesterification but is not further metabolized. M16, but not Cl-DICA, acted as a potent insulin secretagogue in islets cultured in basal but not high glucose. Short-term exposure to M16 or Cl-DICA resulted in activation of glucose- but not arginine-stimulated insulin secretion. Long-term exposure to M16, but not to Cl-DICA, resulted in suppression of glucose-, arginine-, and K(+)-stimulated insulin secretion; inhibition of glucose-induced proinsulin biosynthesis; and depletion of islets insulin. beta Cell mass and islet ATP content remained unaffected. Hence, nonmetabolizable LCFA analogs may highlight discrete LCFA metabolites and pathways involved in modulating insulin secretion, which could be overlooked due to the rapid turnover of natural LCFA. PMID- 17130496 TI - Blockade of pancreatic islet-derived ghrelin enhances insulin secretion to prevent high-fat diet-induced glucose intolerance. AB - The gastric hormone ghrelin and its receptor, growth hormone secretagogue receptor (GHSR), are expressed in pancreas. Here, we report that ghrelin is released from pancreatic islets to regulate glucose-induced insulin release. Plasma concentrations of ghrelin, as well as insulin, were higher in pancreatic veins than in arteries. GHSR antagonist and immunoneutralization of endogenous ghrelin enhanced glucose-induced insulin release from perfused pancreas, whereas exogenous ghrelin suppressed it. GHSR antagonist increased plasma insulin levels in gastrectomized and normal rats to a similar extent. Ghrelin knockout mice displayed enhanced glucose-induced insulin release from isolated islets, whereas islet density, size, insulin content, and insulin mRNA levels were unaltered. Glucose tolerance tests (GTTs) in ghrelin knockout mice showed increased insulin and decreased glucose responses. Treatment with high-fat diet produced glucose intolerance in GTTs in wild-type mice. In ghrelin knockout mice, the high-fat diet-induced glucose intolerance was largely prevented, whereas insulin responses to GTTs were markedly enhanced. These findings demonstrate that ghrelin originating from pancreatic islets is a physiological regulator of glucose induced insulin release. Antagonism of the ghrelin function can enhance insulin release to meet increased demand for insulin in high-fat diet-induced obesity and thereby normalize glycemic control, which may provide a potential therapeutic application to counteract the progression of type 2 diabetes. PMID- 17130497 TI - Separately inherited defects in insulin exocytosis and beta-cell glucose metabolism contribute to type 2 diabetes. AB - The effects of genetic variation on molecular functions predisposing to type 2 diabetes are still largely unknown. Here, in a specifically designed diabetes model, we couple separate gene loci to mechanisms of beta-cell pathology. Niddm1i is a major glucose-controlling 16-Mb region in the diabetic GK rat that causes defective insulin secretion and corresponds to loci in humans and mice associated with type 2 diabetes. Generation of a series of congenic rat strains harboring different parts of GK-derived Niddm1i enabled fine mapping of this locus. Congenic strains carrying the GK genotype distally in Niddm1i displayed reduced insulin secretion in response to both glucose and high potassium, as well as decreased single-cell exocytosis. By contrast, a strain carrying the GK genotype proximally in Niddm1i exhibited both intact insulin release in response to high potassium and intact single-cell exocytosis, but insulin secretion was suppressed when stimulated by glucose. Islets from this strain also failed to respond to glucose by increasing the cellular ATP-to-ADP ratio. Changes in beta-cell mass did not contribute to the secretory defects. We conclude that the failure of insulin secretion in type 2 diabetes includes distinct functional defects in glucose metabolism and insulin exocytosis of the beta-cell and that their genetic fundaments are encoded by different loci within Niddm1i. PMID- 17130498 TI - Peroxisome proliferator-activated receptor-alpha and glucocorticoids interactively regulate insulin secretion during pregnancy. AB - We evaluated the impact of peroxisome proliferator-activated receptor (PPAR)alpha activation and dexamethasone treatment on islet adaptations to the distinct metabolic challenges of fasting and pregnancy, situations where lipid handling is modified to conserve glucose. PPARalpha activation (24 h) in vivo did not affect glucose-stimulated insulin secretion (GSIS) in nonpregnant female rats in the fasted state, although fasting suppressed GSIS. Dexamethasone treatment (5 days) of nonpregnant rats lowered the glucose threshold and augmented GSIS at high glucose; the former effect was selectively opposed by PPARalpha activation. Pregnancy-induced changes in GSIS were opposed by PPARalpha activation at day 19 of pregnancy. Dexamethasone treatment from day 14 to 19 of pregnancy did not modify the GSIS profile of perifused islets from 19-day pregnant rats but rendered the islet GSIS profile refractory to PPARalpha activation. During sustained hyperglycemia in vivo, dexamethasone treatment augmented GSIS in nonpregnant rats but limited further modification of GSIS by pregnancy. We propose that the effect of PPARalpha activation to oppose lowering of the glucose threshold for GSIS by glucocorticoids is important as part of the fasting adaptation, and modulation of the islet GSIS profile by glucocorticoids toward term facilitates the transition of maternal islet function from the metabolic demands of pregnancy to those imposed after parturition. PMID- 17130499 TI - Contribution of calcium influx in mediating glucose-stimulated oxygen consumption in pancreatic islets. AB - In brain, muscle, and pancreatic islets, depolarization induces an increase in respiration, which is dependent on calcium influx. The goal of this study was to assess the quantitative significance of this effect in islets relative to glucose stimulated ATP turnover, to examine the molecular mechanism mediating the changes, and to investigate the functional implications with respect to insulin secretion. Glucose (3-20 mmol/l) increased steady-state levels of cytochrome c reduction (32-66%) in isolated rat islets, reflecting an increased production of NADH, and oxygen consumption rate (OCR) by 0.32 nmol/min/100 islets. Glucose stimulated OCR was inhibited 30% by inhibitors of calcium influx (diazoxide or nimodipine), whereas a protein synthesis inhibitor (emetine) decreased it by only 24%. None of the inhibitors affected cytochrome c reduction, suggesting that calcium's effect on steady-state OCR is mediated by changes in ATP usage rather than the rate of NADH generation. 3-isobutyl-1-methylxanthine increased insulin secretion but had little effect on OCR, indicating that the processes of movement and exocytosis of secretory granules do not significantly contribute to ATP turnover. At 20 mmol/l glucose, a blocker of sarcoendoplasmic reticulum calcium ATPase (SERCA) had little effect on OCR despite a large increase in cytosolic calcium, further supporting the notion that influx of calcium, not bulk cytosolic calcium, is associated with the increase in ATP turnover. The glucose dose response of calcium influx-dependent OCR showed a remarkable correlation with insulin secretion, suggesting that the process mediating the effect of calcium on ATP turnover has a role in the amplification pathway of insulin secretion. PMID- 17130500 TI - Differential effects of p27 in regulation of beta-cell mass during development, neonatal period, and adult life. AB - beta-Cell cycle progression and proliferation are critical to maintain beta-cell mass in adult mice. Of the cell cycle inhibitors, p27Kip1 is thought to be the primary modulator of the proliferative status in most cell types. p27 plays a role in beta-cell adaptation in genetic models of insulin resistance. To study the role of p27 in beta-cells during physiological conditions and at different stages of beta-cell differentiation, we studied mice deficient of or overexpressing p27. Experiments in p27-deficient mice showed improved glucose tolerance and hyperinsulinemia. These changes were associated with increased islet mass and proliferation. The experiments overexpressing p27 in beta-cells were performed using a doxycycline-inducible model. Interestingly, overexpression of p27 for 16 weeks in beta-cells from adult mice had no effect on glucose tolerance, beta-cell mass, or proliferation. In contrast, induction of p27 expression during beta-cell development or early neonatal period resulted in severe glucose intolerance and reduced beta-cell mass by decreased proliferation. These changes were reversible upon discontinuation of doxycycline. These experiments suggest that p27 is a critical molecule for beta-cell proliferation during beta-cell development and early postnatal life but not for maintenance of adult mass. PMID- 17130501 TI - Effects of sex and hormone replacement therapy use on the prevalence of isolated impaired fasting glucose and isolated impaired glucose tolerance in subjects with a family history of type 2 diabetes. AB - Impaired fasting glucose (IFG) is more prevalent in men and impaired glucose tolerance (IGT) more prevalent in women. To explore whether this sex difference is related to female sex hormones, we performed a cross-sectional analysis of data from 2,164 (1,329 women and 835 men) first-degree relatives of individuals with type 2 diabetes. Subjects were categorized based on a 75-g oral glucose tolerance test. Sex and hormone replacement therapy (HRT) effects on the distribution of glucose tolerance were assessed using multinomial logistic regression corrected for familial clustering. Compared with men, women were more likely to have isolated IGT (relative risk 1.8 [95% CI 1.3-2.5]) and less likely to have isolated IFG (0.5 [0.3-0.7]) adjusted for ethnicity, age, waist, fasting insulin, and early insulin release (DeltaI(0-30)/DeltaG(0-30)). To evaluate HRT effects, postmenopausal women using (n = 238) or not using (n = 378) HRT were compared. HRT users were more likely to have isolated IGT (2.2 [1.2-4.0]) after adjustment, but the prevalence of isolated IFG did not differ by HRT status. Based on the influence of sex and HRT on the prevalence of isolated IFG and isolated IGT, we conclude that female sex hormones may play an important role in the pathogenesis of IFG and IGT. PMID- 17130502 TI - Pathogenesis of pre-diabetes: mechanisms of fasting and postprandial hyperglycemia in people with impaired fasting glucose and/or impaired glucose tolerance. AB - Thirty-two subjects with impaired fasting glucose (IFG) and 28 subjects with normal fasting glucose (NFG) ingested a labeled meal and 75 g glucose (oral glucose tolerance test) on separate occasions. Fasting glucose, insulin, and C peptide were higher (P < 0.05) in subjects with IFG than in those with NFG, whereas endogenous glucose production (EGP) did not differ, indicating hepatic insulin resistance. EGP was promptly suppressed, and meal glucose appearance comparably increased following meal ingestion in both groups. In contrast, glucose disappearance (R(d)) immediately after meal ingestion was lower (P < 0.001) in subjects with IFG/impaired glucose tolerance (IGT) and IFG/diabetes but did not differ in subjects with IFG/normal glucose tolerance (NGT) or NFG/NGT. Net insulin action (S(i)) and insulin-stimulated glucose disposal (S(i)*) were reduced (P < 0.001, ANOVA) in subjects with NFG/IGT, IFG/IGT, and IFG/diabetes but did not differ in subjects with NFG/NGT or IFG/NGT. Defective insulin secretion also contributed to lower postprandial R(d) since disposition indexes were lower (P < 0.001, ANOVA) in subjects with NFG/IGT, IFG/IGT, and IFG/diabetes but did not differ in subjects with NFG/NGT and IFG/NGT. We conclude that postprandial hyperglycemia in individuals with early diabetes is due to lower rates of glucose disappearance rather than increased meal appearance or impaired suppression of EGP, regardless of their fasting glucose. In contrast, insulin secretion, action, and the pattern of postprandial turnover are essentially normal in individuals with isolated IFG. PMID- 17130503 TI - Irbesartan treatment reduces biomarkers of inflammatory activity in patients with type 2 diabetes and microalbuminuria: an IRMA 2 substudy. AB - The impact of irbesartan treatment on biomarkers of low-grade inflammation, endothelial dysfunction, growth factors, and advanced glycation end products (AGEs) during the Irbesartan in Patients with Type 2 Diabetes and Microalbuminuria (IRMA 2) study was evaluated. IRMA 2 was a 2-year multicenter, randomized, double-blind trial in patients comparing irbesartan (150 or 300 mg once daily) versus placebo. The primary end point was onset of overt nephropathy. A subgroup (n = 269, 68%) was analyzed for biomarkers at baseline and after 1 and 2 years. High-sensitivity C-reactive protein (hs-CRP), interleukin (IL)-6, fibrinogen, adhesion molecules, transforming growth factor-beta, and AGE peptides were assessed. Irbesartan treatment yielded significant changes in hs-CRP (based on generalized estimating equation regression coefficient) with a 5.4% decrease per year versus a 10% increase per year in the placebo group (P < 0.001). Fibrinogen decreased 0.059 g/l per year from baseline versus placebo's 0.059 g/l increase per year (P = 0.027). IL-6 showed a 1.8% increase per year compared with placebo's 6.5% increase per year (P = 0.005). Changes in IL-6 were associated with changes in albumin excretion (P = 0.04). There was no treatment effect on the other biomarkers. Irbesartan (300 mg once daily) reduces low-grade inflammation in this high-risk population, and this may reduce the risk of micro- and macrovascular disease. PMID- 17130504 TI - The effect of intensive glycemic treatment on coronary artery calcification in type 1 diabetic participants of the Diabetes Control and Complications Trial/Epidemiology of Diabetes Interventions and Complications (DCCT/EDIC) Study. AB - The Epidemiology of Diabetes Interventions and Complications (EDIC) study, an observational follow-up of the Diabetes Control and Complications Trial (DCCT) type 1 diabetes cohort, measured coronary artery calcification (CAC), an index of atherosclerosis, with computed tomography (CT) in 1,205 EDIC patients at approximately 7-9 years after the end of the DCCT. We examined the influence of the 6.5 years of prior conventional versus intensive diabetes treatment during the DCCT, as well as the effects of cardiovascular disease risk factors, on CAC. The prevalences of CAC >0 and >200 Agatston units were 31.0 and 8.5%, respectively. Compared with the conventional treatment group, the intensive group had significantly lower geometric mean CAC scores and a lower prevalence of CAC >0 in the primary retinopathy prevention cohort, but not in the secondary intervention cohort, and a lower prevalence of CAC >200 in the combined cohorts. Waist-to-hip ratio, smoking, hypertension, and hypercholesterolemia, before or at the time of CT, were significantly associated with CAC in univariate and multivariate analyses. CAC was associated with mean HbA(1c) (A1C) levels before enrollment, during the DCCT, and during the EDIC study. Prior intensive diabetes treatment during the DCCT was associated with less atherosclerosis, largely because of reduced levels of A1C during the DCCT. PMID- 17130505 TI - Effect of eucaloric high- and low-sucrose diets with identical macronutrient profile on insulin resistance and vascular risk: a randomized controlled trial. AB - The long-term impact of dietary carbohydrate type, in particular sucrose, on insulin resistance and the development of diabetes and atherosclerosis is not established. Current guidelines for the healthy population advise restriction of sucrose intake. We investigated the effect of high- versus low-sucrose diet (25 vs. 10%, respectively, of total energy intake) in 13 healthy subjects aged 33 +/- 3 years (mean +/- SE), BMI 26.6 +/- 0.9 kg/m(2), in a randomized crossover design with sequential 6-week dietary interventions separated by a 4-week washout. Weight maintenance, eucaloric diets with identical macronutrient profiles and fiber content were designed. All food was weighed and distributed. Insulin action was assessed using a two-step euglycemic clamp; glycemic profiles were assessed by the continuous glucose monitoring system and vascular compliance by pulse-wave analysis. There was no change in weight across the study. Peripheral glucose uptake and suppression of endogenous glucose production were similar after each diet. Glycemic profiles and measures of vascular compliance did not change. A rise in total and LDL cholesterol was observed. In this study, a high-sucrose intake as part of an eucaloric, weight-maintaining diet had no detrimental effect on insulin sensitivity, glycemic profiles, or measures of vascular compliance in healthy nondiabetic subjects. PMID- 17130506 TI - Diabetes alters sphingolipid metabolism in the retina: a potential mechanism of cell death in diabetic retinopathy. AB - Dysregulated sphingolipid metabolism causes neuronal cell death and is associated with insulin resistance and diseases. Thus, we hypothesized that diabetes-induced changes in retinal sphingolipid metabolism may contribute to neuronal pathologies in diabetic retinopathy. ESI-MS/MS was used to measure ceramide content and ceramide metabolites in whole retinas after 2, 4, and 8 weeks of streptozotocin induced diabetes. After 4 and 8 weeks of diabetes, a approximately 30% decrease in total ceramide content was observed, concomitant with a significant approximately 30% increase in glucosylceramide levels in fed diabetic rats compared with their age-matched controls. Acute insulin therapy as well as a short-term lowering of glucose via fasting did not affect the increase in glucosylceramide composition. To assess the putative biological consequences of the increase in glucosylceramide composition, R28 retinal neurons were treated with glucosylceramide synthase inhibitors. Inhibiting glycosphingolipid metabolism increased insulin sensitivity in retinal neurons. Glycosphingolipid inhibitors augmented insulin-stimulated p70 S6kinase activity in the presence of inhibitory concentrations of high glucose or glucosamine. Inhibition of glycosphingolipid synthesis also suppressed glucosamine- and interleukin-1beta induced death. Consistent with these inhibitor studies, pharmacological accumulation of glycosphingolipids increased activation of the endoplasmic reticulum stress response, a putative modulator of insulin resistance and neuronal apoptosis. It is speculated that an increase in glucosylceramide, and possibly higher-order glycosphingolipids, could contribute to the pathogenesis of diabetic retinopathy by contributing to local insulin resistance, resulting in neuronal cell death. Thus, dysfunctional glycosphingolipid metabolism may contribute to metabolic stress in diabetes, and therapeutic strategies to restore normal sphingolipid metabolism may be a viable approach for treatment of diabetic retinopathy. PMID- 17130507 TI - C-Peptide reverses nociceptive neuropathy in type 1 diabetes. AB - We examined the therapeutic effects of C-peptide on established nociceptive neuropathy in type 1 diabetic BB/Wor rats. Nociceptive nerve function, unmyelinated sural nerve fiber and dorsal root ganglion (DRG) cell morphometry, nociceptive peptide content, and the expression of neurotrophic factors and their receptors were investigated. C-peptide was administered either as a continuous subcutaneous replacement dose via osmopumps or a replacement dose given once daily by subcutaneous injection. Diabetic rats were treated from 4 to 7 months of diabetes and were compared with control and untreated diabetic rats of 4- and 7 month duration. Osmopump delivery but not subcutaneous injection improved hyperalgesia and restored the diabetes-induced reduction of unmyelinated fiber number (P < 0.01) and mean axonal size (P < 0.05) in the sural nerve. High affinity nerve growth factor (NGF) receptor (NGFR-TrkA) expression in DRGs was significantly reduced at 4 months (P < 0.01). Insulin receptor and IGF-I receptor (IGF-IR) expressions in DRGs and NGF content in sciatic nerve were significantly decreased in 7-month diabetic rats (P < 0.01, 0.05, and 0.005, respectively). Osmopump delivery prevented the decline of NGFR-TrkA, insulin receptor (P < 0.05), and IGF-IR (P < 0.005) expressions in DRGs and improved NGF content (P < 0.05) in sciatic nerve. However, subcutaneous injection had only marginal effects on morphometric and molecular changes in diabetic rats. We conclude that C peptide exerts beneficial therapeutic effects on diabetic nociceptive neuropathy and that optimal effects require maintenance of physiological C-peptide concentrations for a major proportion of the day. PMID- 17130508 TI - Reduced expression of G(i) in erythrocytes of humans with type 2 diabetes is associated with impairment of both cAMP generation and ATP release. AB - Human erythrocytes, by virtue of their ability to release ATP in response to physiological stimuli, have been proposed to participate in the regulation of local blood flow. A signal transduction pathway that relates these stimuli to ATP release has been described and includes the heterotrimeric G protein G(i) and adenylyl cyclase (AC). In this cell, G(i) activation results in increases in cAMP and, ultimately, ATP release. It has been reported that G(i) expression is decreased in animal models of diabetes and in platelets of humans with type 2 diabetes. Here, we report that G(i2) expression is selectively decreased in erythrocytes of humans with type 2 diabetes and that this defect is associated with reductions in cAMP accumulation and ATP release in response to incubation of erythrocytes with mastoparan 7 (10 micromol/l), an activator of G(i). Importantly, this defect in ATP release correlates inversely with the adequacy of glycemic control as determined by levels of HbA(1c) (A1C). These results demonstrate that in erythrocytes of humans with type 2 diabetes, both G(i) expression and ATP release in response to mastoparan 7 are impaired, which is consistent with the hypothesis that this defect in erythrocyte physiology could contribute to the vascular disease associated with this clinical condition. PMID- 17130509 TI - Treatment of spontaneously hypertensive rats with rosiglitazone and/or enalapril restores balance between vasodilator and vasoconstrictor actions of insulin with simultaneous improvement in hypertension and insulin resistance. AB - Spontaneously hypertensive rats (SHRs) exhibit endothelial dysfunction and insulin resistance. Reciprocal relationships between endothelial dysfunction and insulin resistance may contribute to hypertension by causing imbalanced regulation of endothelial-derived vasodilators (e.g., nitric oxide) and vasoconstrictors (e.g., endothelin-1 [ET-1]). Treatment of SHRs with rosiglitazone (insulin sensitizer) and/or enalapril (ACE inhibitor) may simultaneously improve hypertension, insulin resistance, and endothelial dysfunction by rebalancing insulin-stimulated production of vasoactive mediators. When compared with WKY control rats, 12-week-old vehicle-treated SHRs were hypertensive, overweight, and insulin resistant, with elevated fasting levels of insulin and ET-1 and reduced serum adiponectin levels. In mesenteric vascular beds (MVBs) isolated from vehicle-treated SHRs and preconstricted with norepinephrine (NE) ex vivo, vasodilator responses to insulin were significantly impaired, whereas the ability of insulin to oppose vasoconstrictor actions of NE was absent (versus WKY controls). Three-week treatment of SHRs with rosiglitazone and/or enalapril significantly reduced blood pressure, insulin resistance, fasting insulin, and ET-1 levels and increased adiponectin levels to values comparable with those observed in vehicle-treated WKY controls. By restoring phosphatidylinositol 3-kinase-dependent effects, rosiglitazone and/or enalapril therapy of SHRs also significantly improved vasodilator responses to insulin in MVB preconstricted with NE ex vivo. Taken together, our data provide strong support for the existence of reciprocal relationships between endothelial dysfunction and insulin resistance that may be relevant for developing novel therapeutic strategies for the metabolic syndrome. PMID- 17130510 TI - Inhalation of human insulin (exubera) augments the efficiency of muscle glucose uptake in vivo. AB - This study assessed the site of increased glucose uptake resulting from insulin inhalation, quantified its effect under steady-state glucose concentrations, and identified the time to onset of effect. Human insulin was administered to 13 beagles via inhalation (Exubera [insulin human (rDNA origin)] Inhalation Powder; n = 7) or infusion into the inferior vena cava (Humulin R; n = 6) using an algorithm to match plasma insulin levels and kinetics for both groups. Somatostatin and glucagon were infused. Glucose was delivered into the portal vein (4 mg x kg(-1) x min(-1)) and a peripheral vein, as needed, to maintain arterial plasma glucose levels at 180 mg/dl. Hepatic exposure to insulin and glucose and liver glucose uptake were similar in both groups. Despite comparable arterial insulin and glucose levels, hind-limb glucose uptake increased 2.4-fold after inhalation compared with infusion due to increased muscle glucose uptake. Glucose infusion rate, nonhepatic glucose uptake, and tracer-determined glucose disposal were about twice as great compared with intravenous insulin. The effect appeared after 1 h, persisting at least as long as arterial insulin levels remained above basal. Pulmonary administration of insulin increases nonhepatic glucose uptake compared with infusion, and skeletal muscle is the likely site of that effect. PMID- 17130511 TI - The idd4 locus displays sex-specific epistatic effects on type 1 diabetes susceptibility in nonobese diabetic mice. AB - The nonobese diabetic (NOD) mouse recapitulates many aspects of the pathogenesis of type 1 diabetes in humans, including inheritance as a complex trait. More than 20 Idd loci have been linked to type 1 diabetes susceptibility in NOD mice. Previously, we used linkage analysis of NOD crossed to the nonobese diabetes resistant (NOR) strain and NOD congenic strains to map susceptibility to both spontaneous and cyclophosphamide-accelerated type 1 diabetes to the Idd4 locus on chromosome 11 that displayed a sex-specific effect on diabetes susceptibility. Here, we elucidate the complex genetic architecture of Idd4 by analysis of congenic strains on the NOD and NOR backgrounds. We previously refined Idd4.1 to 1.4 Mb and demonstrated an impact of this interval on type 1 interferon pathways in antigen-presenting cells. Here, we identify a second subregion, the 0.92 Mb Idd4.2 locus located telomeric to Idd4.1. Strikingly, Idd4.2 displayed a sex specific, epistatic interaction with Idd4.1 in NOR.NOD congenic females that was not observed in syngenic males. Idd4.2 contains 29 genes, and promising candidates for the Idd4.2 effect on type 1 diabetes are described. These data demonstrate sex-dependent interaction effects on type 1 diabetes susceptibility and provide a framework for functional analysis of Idd4.2 candidate genes. PMID- 17130512 TI - The Kruppel-like factor 11 (KLF11) Q62R polymorphism is not associated with type 2 diabetes in 8,676 people. AB - Kruppel-like factor 11 is a pancreatic transcription factor whose activity induces the insulin gene. A common glutamine-to-arginine change at codon 62 (Q62R) in its gene KLF11 has been recently associated with type 2 diabetes in two independent samples. Q62R and two other rare missense variants (A347S and T220M) were also shown to affect the function of KLF11 in vitro, and insulin levels were lower in carriers of the minor allele at Q62R. We therefore examined their impact on common type 2 diabetes in several family-based and case-control samples of northern-European ancestry, totaling 8,676 individuals. We did not detect the rare A347S and T220M variants in our samples. With respect to Q62R, despite >99% power to detect an association of the previously published magnitude, Q62R was not associated with type 2 diabetes (pooled odds ratio 0.97 [95% CI 0.88-1.08], P = 0.63). In a subset of normoglycemic individuals, we did not observe significant differences in various insulin traits according to genotype at KLF11 Q62R. We conclude that the KLF11 A347S and T220M mutations do not contribute to increased risk of diabetes in European-derived populations and that the Q62R polymorphism has, at best, a minor effect on diabetes risk. PMID- 17130513 TI - CHRM3 gene variation is associated with decreased acute insulin secretion and increased risk for early-onset type 2 diabetes in Pima Indians. AB - The muscarinic acetylcholine receptor subtype M3 (CHRM3) gene is expressed in islet beta-cells and has a role in stimulating insulin secretion; therefore, CHRM3 was analyzed as a candidate gene for type 2 diabetes in Pima Indians. Ten variants were genotyped in a family-based sample (n = 1,037), and 1 variant (rs3738435) located in the 5' untranslated region of an alternative transcript was found to be modestly associated with both early-onset type 2 diabetes and the acute insulin response in a small subset of these subjects. To better assess whether this variant has a role in acute insulin secretion, which could affect risk for early-onset type 2 diabetes, rs3738435 was genotyped in a larger group of normal glucose-tolerant Pima Indians who had measures of acute insulin secretion (n = 282) and a larger case-control group of Pima Indians selected for early-onset type 2 diabetes (n = 348 case subjects with age of onset <25 years; n = 392 nondiabetic control subjects aged >45 years). Genotyping in these larger sets of subjects confirmed that the C allele of rs3738435 was associated with a reduced acute insulin response (adjusted P = 0.00006) and was also modestly associated with increased risk of early-onset type 2 diabetes (adjusted P = 0.02). PMID- 17130514 TI - Polymorphism in the transcription factor 7-like 2 (TCF7L2) gene is associated with reduced insulin secretion in nondiabetic women. AB - Recently, the transcription factor 7-like 2 (TCF7L2) gene on chromosome 10q25.2 has been linked with type 2 diabetes among Caucasians, with disease associations noted for single nucleotide polymorphisms (SNPs) rs12255372 and rs7903146. To investigate mechanisms by which TCF7L2 could contribute to type 2 diabetes, we examined the effects of these SNPs on clinical and metabolic traits affecting glucose homeostasis in 256 nondiabetic female subjects (138 European Americans and 118 African Americans) aged 7-57 years. Outcomes included BMI, percent body fat, insulin sensitivity (S(i)), acute insulin response to glucose (AIR(g)), and the disposition index (DI). Homozygosity for the minor allele (TT) of SNP rs12255372 occurred in 9% of individuals and was associated with a 31% reduction in DI values in a recessive model. The at-risk allele TT was also associated with lower AIR(g) adjusted for S(i) in both ethnic groups, whereas rs12255372 genotype was not associated with measures of adiposity or with S(i). The T allele of rs12255372 was also associated with increased prevalence of impaired fasting glucose. Genotypes at rs7903146 were not associated with any metabolic trait. Lower S(i) and higher AIR(g) observed in the African-American compared with the European-American subgroup could not be explained by the TCF7L2 genotype. Our data suggest that the TCF7L2 gene is an important factor regulating insulin secretion, which could explain its association with type 2 diabetes. PMID- 17130515 TI - Amenorrhea in premenopausal women after adjuvant chemotherapy for breast cancer. AB - Chemotherapy and ovarian ablation both independently improve survival in premenopausal women with hormone-sensitive breast cancer. Amenorrhea is a well recognized occurrence after chemotherapy. The rate of chemotherapy-induced amenorrhea varies with patient age and chemotherapy regimens administered. However, the impact of chemotherapy-induced amenorrhea on prognosis is still being defined. Older studies in premenopausal women argue that the benefit with chemotherapy is a result of direct cytotoxicity alone. However, studies that restrict outcome analysis to hormone receptor-positive tumors suggest that chemotherapy has a dual mechanism in women with hormone-responsive tumors; indirect endocrine manipulation secondary to chemotherapy-induced ovarian suppression and direct cytotoxicity. The significant health ramifications involved with the induction of premature menopause as well as potential benefits necessitate a comprehensive evaluation of chemotherapy-induced amenorrhea. This review will discuss the incidence of amenorrhea with commonly-used adjuvant chemotherapeutic regimens, the possible benefits of chemotherapy-induced amenorrhea, and the challenges of interpreting the existing data in breast cancer trials. PMID- 17130516 TI - Amyloid ion channels: a porous argument or a thin excuse? PMID- 17130517 TI - Extracellular blockade of potassium channels by TEA+: the tip of the iceberg? PMID- 17130518 TI - A cation-pi interaction between extracellular TEA and an aromatic residue in potassium channels. AB - Open-channel blockers such as tetraethylammonium (TEA) have a long history as probes of the permeation pathway of ion channels. High affinity blockade by extracellular TEA requires the presence of an aromatic amino acid at a position that sits at the external entrance of the permeation pathway (residue 449 in the eukaryotic voltage-gated potassium channel Shaker). We investigated whether a cation-pi interaction between TEA and such an aromatic residue contributes to TEA block using the in vivo nonsense suppression method to incorporate a series of increasingly fluorinated Phe side chains at position 449. Fluorination, which is known to decrease the cation-pi binding ability of an aromatic ring, progressively increased the inhibitory constant K(i) for the TEA block of Shaker. A larger increase in K(i) was observed when the benzene ring of Phe449 was substituted by nonaromatic cyclohexane. These results support a strong cation-pi component to the TEA block. The data provide an empirical basis for choosing between Shaker models that are based on two classes of reported crystal structures for the bacterial channel KcsA, showing residue Tyr82 in orientations either compatible or incompatible with a cation-pi mechanism. We propose that the aromatic residue at this position in Shaker is favorably oriented for a cation-pi interaction with the permeation pathway. This choice is supported by high level ab initio calculations of the predicted effects of Phe modifications on TEA binding energy. PMID- 17130519 TI - Dynamic inositol trisphosphate-mediated calcium signals within astrocytic endfeet underlie vasodilation of cerebral arterioles. AB - Active neurons communicate to intracerebral arterioles in part through an elevation of cytosolic Ca(2+) concentration ([Ca(2+)](i)) in astrocytes, leading to the generation of vasoactive signals involved in neurovascular coupling. In particular, [Ca(2+)](i) increases in astrocytic processes ("endfeet"), which encase cerebral arterioles, have been shown to result in vasodilation of arterioles in vivo. However, the spatial and temporal properties of endfoot [Ca(2+)](i) signals have not been characterized, and information regarding the mechanism by which these signals arise is lacking. [Ca(2+)](i) signaling in astrocytic endfeet was measured with high spatiotemporal resolution in cortical brain slices, using a fluorescent Ca(2+) indicator and confocal microscopy. Increases in endfoot [Ca(2+)](i) preceded vasodilation of arterioles within cortical slices, as detected by simultaneous measurement of endfoot [Ca(2+)](i) and vascular diameter. Neuronal activity-evoked elevation of endfoot [Ca(2+)](i) was reduced by inhibition of inositol 1,4,5-trisphosphate (InsP(3)) receptor Ca(2+) release channels and almost completely abolished by inhibition of endoplasmic reticulum Ca(2+) uptake. To probe the Ca(2+) release mechanisms present within endfeet, spatially restricted flash photolysis of caged InsP(3) was utilized to liberate InsP(3) directly within endfeet. This maneuver generated large amplitude [Ca(2+)](i) increases within endfeet that were spatially restricted to this region of the astrocyte. These InsP(3)-induced [Ca(2+)](i) increases were sensitive to depletion of the intracellular Ca(2+) store, but not to ryanodine, suggesting that Ca(2+)-induced Ca(2+) release from ryanodine receptors does not contribute to the generation of endfoot [Ca(2+)](i) signals. Neuronally evoked increases in astrocytic [Ca(2+)](i) propagated through perivascular astrocytic processes and endfeet as multiple, distinct [Ca(2+)](i) waves and exhibited a high degree of spatial heterogeneity. Regenerative Ca(2+) release processes within the endfeet were evident, as were localized regions of Ca(2+) release, and treatment of slices with the vasoactive neuropeptides somatostatin and vasoactive intestinal peptide was capable of inducing endfoot [Ca(2+)](i) increases, suggesting the potential for signaling between local interneurons and astrocytic endfeet in the cortex. Furthermore, photorelease of InsP(3) within individual endfeet resulted in a local vasodilation of adjacent arterioles, supporting the concept that astrocytic endfeet function as local "vasoregulatory units" by translating information from active neurons into complex InsP(3)-mediated Ca(2+) release signals that modulate arteriolar diameter. PMID- 17130520 TI - Conformational dynamics of hSGLT1 during Na+/glucose cotransport. AB - This study examines the conformations of the Na(+)/glucose cotransporter (SGLT1) during sugar transport using charge and fluorescence measurements on the human SGLT1 mutant G507C expressed in Xenopus oocytes. The mutant exhibited similar steady-state and presteady-state kinetics as wild-type SGLT1, and labeling of Cys507 by tetramethylrhodamine-6-maleimide had no effect on kinetics. Our strategy was to record changes in charge and fluorescence in response to rapid jumps in membrane potential in the presence and absence of sugar or the competitive inhibitor phlorizin. In Na(+) buffer, step jumps in membrane voltage elicited presteady-state currents (charge movements) that decay to the steady state with time constants tau(med) (3-20 ms, medium) and tau(slow) (15-70 ms, slow). Concurrently, SGLT1 rhodamine fluorescence intensity increased with depolarizing and decreased with hyperpolarizing voltages (DeltaF). The charge vs. voltage (Q-V) and fluorescence vs. voltage (DeltaF-V) relations (for medium and slow components) obeyed Boltzmann relations with similar parameters: zdelta (apparent valence of voltage sensor) approximately 1; and V(0.5) (midpoint voltage) between -15 and -40 mV. Sugar induced an inward current (Na(+)/glucose cotransport), and reduced maximal charge (Q(max)) and fluorescence (DeltaF(max)) with half-maximal concentrations (K(0.5)) of 1 mM. Increasing [alphaMDG](o) also shifted the V(0.5) for Q and DeltaF to more positive values, with K(0.5)'s approximately 1 mM. The major difference between Q and DeltaF was that at saturating [alphaMDG](o), the presteady-state current (and Q(max)) was totally abolished, whereas DeltaF(max) was only reduced 50%. Phlorizin reduced both Q(max) and DeltaF(max) (K(i) approximately 0.4 microM), with no changes in V(0.5)'s or relaxation time constants. Simulations using an eight-state kinetic model indicate that external sugar increases the occupancy probability of inward facing conformations at the expense of outward-facing conformations. The simulations predict, and we have observed experimentally, that presteady-state currents are blocked by saturating sugar, but not the changes in fluorescence. Thus we have isolated an electroneutral conformational change that has not been previously described. This rate-limiting step at maximal inward Na(+)/sugar cotransport (saturating voltage and external Na(+) and sugar concentrations) is the slow release of Na(+) from the internal surface of SGLT1. The high affinity blocker phlorizin locks the cotransporter in an inactive conformation. PMID- 17130521 TI - Secondary structure of a KCNE cytoplasmic domain. AB - Type I transmembrane KCNE peptides contain a conserved C-terminal cytoplasmic domain that abuts the transmembrane segment. In KCNE1, this region is required for modulation of KCNQ1 K(+) channels to afford the slowly activating cardiac I(Ks) current. We utilized alanine/leucine scanning to determine whether this region possesses any secondary structure and to identify the KCNE1 residues that face the KCNQ1 channel complex. Helical periodicity analysis of the mutation induced perturbations in voltage activation and deactivation kinetics of KCNQ1 KCNE1 complexes defined that the KCNE1 C terminus is alpha-helical when split in half at a conserved proline residue. This helical rendering assigns all known long QT mutations in the KCNE1 C-terminal domain as protein facing. The identification of a secondary structure within the KCNE1 C-terminal domain provides a structural scaffold to map protein-protein interactions with the pore forming KCNQ1 subunit as well as the cytoplasmic regulatory proteins anchored to KCNQ1-KCNE complexes. PMID- 17130522 TI - An S6 mutation in BK channels reveals beta1 subunit effects on intrinsic and voltage-dependent gating. AB - Large conductance, Ca(2+)- and voltage-activated K(+) (BK) channels are exquisitely regulated to suit their diverse roles in a large variety of physiological processes. BK channels are composed of pore-forming alpha subunits and a family of tissue-specific accessory beta subunits. The smooth muscle specific beta1 subunit has an essential role in regulating smooth muscle contraction and modulates BK channel steady-state open probability and gating kinetics. Effects of beta1 on channel's gating energetics are not completely understood. One of the difficulties is that it has not yet been possible to measure the effects of beta1 on channel's intrinsic closed-to-open transition (in the absence of voltage sensor activation and Ca(2+) binding) due to the very low open probability in the presence of beta1. In this study, we used a mutation of the alpha subunit (F315Y) that increases channel openings by greater than four orders of magnitude to directly compare channels' intrinsic open probabilities in the presence and absence of the beta1 subunit. Effects of beta1 on steady-state open probabilities of both wild-type alpha and the F315Y mutation were analyzed using the dual allosteric HA model. We found that mouse beta1 has two major effects on channel's gating energetics. beta1 reduces the intrinsic closed-to open equilibrium that underlies the inhibition of BK channel opening seen in submicromolar Ca(2+). Further, P(O) measurements at limiting slope allow us to infer that beta1 shifts open channel voltage sensor activation to negative membrane potentials, which contributes to enhanced channel opening seen at micromolar Ca(2+) concentrations. Using the F315Y alpha subunit with deletion mutants of beta1, we also demonstrate that the small N- and C-terminal intracellular domains of beta1 play important roles in altering channel's intrinsic opening and voltage sensor activation. In summary, these results demonstrate that beta1 has distinct effects on BK channel intrinsic gating and voltage sensor activation that can be functionally uncoupled by mutations in the intracellular domains. PMID- 17130523 TI - A dipeptidyl aminopeptidase-like protein remodels gating charge dynamics in Kv4.2 channels. AB - Dipeptidyl aminopeptidase-like proteins (DPLPs) interact with Kv4 channels and thereby induce a profound remodeling of activation and inactivation gating. DPLPs are constitutive components of the neuronal Kv4 channel complex, and recent observations have suggested the critical functional role of the single transmembrane segment of these proteins (Zagha, E., A. Ozaita, S.Y. Chang, M.S. Nadal, U. Lin, M.J. Saganich, T. McCormack, K.O. Akinsanya, S.Y. Qi, and B. Rudy. 2005. J. Biol. Chem. 280:18853-18861). However, the underlying mechanism of action is unknown. We hypothesized that a unique interaction between the Kv4.2 channel and a DPLP found in brain (DPPX-S) may remodel the channel's voltage sensing domain. To test this hypothesis, we implemented a robust experimental system to measure Kv4.2 gating currents and study gating charge dynamics in the absence and presence of DPPX-S. The results demonstrated that coexpression of Kv4.2 and DPPX-S causes a -26 mV parallel shift in the gating charge-voltage (Q V) relationship. This shift is associated with faster outward movements of the gating charge over a broad range of relevant membrane potentials and accelerated gating charge return upon repolarization. In sharp contrast, DPPX-S had no effect on gating charge movements of the Shaker B Kv channel. We propose that DPPX-S destabilizes resting and intermediate states in the voltage-dependent activation pathway, which promotes the outward gating charge movement. The remodeling of gating charge dynamics may involve specific protein-protein interactions of the DPPX-S's transmembrane segment with the voltage-sensing and pore domains of the Kv4.2 channel. This mechanism may determine the characteristic fast operation of neuronal Kv4 channels in the subthreshold range of membrane potentials. PMID- 17130524 TI - Overexpression of ER and VDR is not sufficient to make ER-negative MDA-MB231 breast cancer cells responsive to 1alpha-hydroxyvitamin D5. AB - 1alpha-hydroxyvitamin D(5) [1alpha(OH)D(5)] is an active vitamin D analog showing promising chemopreventive effect in breast carcinogenesis. We previously reported that estrogen receptor (ER)-positive breast cancer cells were sensitive, whereas ER-negative breast cancer cells were relatively resistant to their antiproliferative effects. In the present study, we used ER-negative MDA-MB231, ER-transfected MDA-MB231 (S30) and ER-positive BT474 cell lines to evaluate the possible association between ER status and cellular sensitivity to 1alpha(OH)D(5) treatment. Our results demonstrate that ER expression in ER-negative breast cancer cells (S30) did not increase the sensitivity to 1alpha(OH)D(5), whereas in ER-positive BT474 cells, the significant antiproliferative effect of 1alpha(OH)D(5) was correlated with the downregulation of ER and progesterone receptor expression. Further analysis indicated that both MDA-MB231 and S30 cells express low vitamin D receptor (VDR) at transcriptional level and protein level. However, transfection of VDR failed to restore the sensitivity to 1alpha(OH)D(5) in MDA-MB231 and S30 cells, although VDR direct target gene CYP24 was more responsive to 1alpha(OH)D(5) treatment in MDA-MB231 and S30 cells overexpressing VDR. In addition, nuclear receptor cofactors NCoR1 and SRC1 that could potentially affect VDR action were also low in both MDA-MB231 and S30 cells in comparison with ER-positive, vitamin D-sensitive BT474 cells. These results suggest that in addition to the increased ER and VDR expression, the intact VDR signaling machinery as present in ER-positive, vitamin D-sensitive cells is essential for the antiproliferative action of vitamin D, whereas the direct VDR target genes such as CYP24 can remain responsive to augmented VDR expression. PMID- 17130525 TI - The Type 1 Diabetes Genetics Consortium. AB - The Type 1 Diabetes Genetics Consortium (T1DGC) is an international, multicenter program organized to promote research to identify genes and their alleles that determine an individual's risk for type 1 diabetes (T1D). The primary goal of the T1DGC is to establish resources and data that can be used by, and that is fully accessible to, the research community in the study of T1D. All the information on T1DGC can be accessed at the following web address: http://www.t1dgc.org. A resource base of well-characterized families is being assembled that will facilitate the localization and characterization of T1D susceptibility genes. From these families, the T1DGC is establishing banks of DNA, serum, plasma, and cell lines, as well as useful databases. The T1DGC also sponsors training opportunities (bioinformatics) and technology transfer (HLA genotyping). PMID- 17130526 TI - A mini meta-analysis of studies on CD4+CD25+ T cells in human type 1 diabetes: report of the Immunology of Diabetes Society T Cell Workshop. AB - Type 1 diabetes mellitus (T1DM) is characterized by a loss of self-tolerance to islet antigens. In health, immunological tolerance is maintained by multiple central and peripheral mechanisms including the action of a specialized set of regulatory T cells characterized by expression of CD4 and CD25 (CD4+CD25+ Treg). It has been suggested that a defect in this cell population, either numerically or functionally, could contribute to the development of autoimmune diseases, such as T1DM. To investigate this possibility, several research groups have studied the frequency and suppressive capacity of this cell population in individuals with T1DM and, to date, there are four such studies published. We therefore performed a mini meta-analysis to compare the results in the four published studies, account for differences in their findings, and draw a consensus view on the role of this important cell subset in human T1DM. PMID- 17130527 TI - HLA class I epitope discovery in type 1 diabetes: independent and reproducible identification of proinsulin epitopes of CD8 T cells--report of the IDS T Cell Workshop Committee. AB - Islet autoreactive CD8 T cells are plausible candidates for direct beta cell toxicity in type 1 diabetes (T1DM). In 2005, cellular studies in the pathogenesis of this disease have reached a new milestone. Autoreactive CD8 T cells have been defined and several target islet epitopes of these have been discovered and validated simultaneously in three independent studies. The insulin B10-B18 peptide that displays exceptional binding affinity for HLA-A2 has been reported in all three studies, and its recognition shows an association with autoimmune beta cell destruction and T1DM. These studies imply that CD8 T cell-based HLA tetramers and ELISPOT analyses can be useful to monitor T1DM as well as islet transplantation, and may provide useful tools to assess immunological efficacy of immune intervention trials. PMID- 17130528 TI - Genetics of fulminant type 1 diabetes. AB - Fulminant type 1 diabetes exhibits distinct clinical futures from "classic" autoimmune type 1 diabetes. Although the etiology of fulminant type 1 diabetes is not fully elucidated, class II HLA could contribute to the development of fulminant type 1 diabetes. In Japanese patients with "classic" type 1 diabetes, DRB1*0405-DQB1*0401 and DRB1*0901-DQB1*0303 are major susceptible HLA-DR-DQ haplotypes, whereas DRB1*1502-DQB1*0601 and DRB1*1501-DQB1*0602 are protective. In contrast, only DRB1*0405-DQB1*0401, but not DRB1*0901-DQB1*0303, is a susceptible haplotype in fulminant type 1 diabetes. In addition, neither DRB1*1502-DQB1*0601 nor DRB1*1501-DQB1*0602 are protective haplotypes in fulminant type 1 diabetes. In genotypic combination analysis, the homozygotes of DRB1*0405-DQB1*0401 are associated with both fulminant type 1 diabetes and "classic" type 1 diabetes, whereas the homozygotes of DRB1*0901-DQB1*0303 are associated with only "classic" type 1 diabetes. These findings suggest a different contribution of class II HLA in the mechanisms of beta cell damage between fulminant and "classic" type 1 diabetes. To further address the pathogenesis of fulminant type 1 diabetes, HNF-1alpha gene mutation and mutation of the mitochondrial DNA were analyzed in patients with fulminant type 1 diabetes admitted to our department during the period from 1990 to 2000. Neither mutations of HNF-1alpha gene nor A-to-G mutation at nucleotide position 3,243 of the mitochondrial tRNA(LEU(UUR)) gene were identified in these patients. These results suggest that the HNF-1alpha gene mutation and mutation of the mitochondrial DNA are not likely associated with diabetic patients with fulminant clinical symptoms at disease onset. In this article we will summarize the current findings on the genetics of Japanese patients with fulminant type 1 diabetes. PMID- 17130529 TI - Why is type 1 diabetes uncommon in Asia? AB - T1D (type 1 diabetes) incidence rates are extremely low in Asian populations. The prevalences of islet-specific autoantibodies are reported to be low compared with Caucasians. Although the clinical and immunologic characteristics of T1D in Asians appear to be different from those of Caucasians, if we apply correct patient definition and standardized methods, the typical T1D patients are very similar, in the immunologic as well as genetic perspectives. Although the association of individual allele seems to be different between populations, if we compare the identical DR-DQ haplotypes, the association and transmission to diabetic offspring were similar for Asians and Caucasians. The high-risk HLA genotypes/haplotypes were found to be independent determinants of diabetes in the first-degree relatives of individuals with T1D, particularly in the presence of autoantibodies. A different genetic susceptibility including a low frequency of high-risk HLA alleles could explain the lower prevalence of islet-specific autoantibodies and the low incidence of T1D, or different genetic and environmental interactions might be involved in the etiology of T1D. It is certain that DR-DQ linkage disequilibrium (LD) is an important factor explaining the difference in T1D incidence in different countries. LD between highly susceptible DRB1 alleles and protective DQB1 alleles, and vice versa, is the major contributing factor to the low incidence of T1D in Asians. We also suggested that different genetic/environmental interactions might operate in the etiology of T1D between Caucasians and Asians. It would be of great help for primary prevention to investigate to what degree genetic determinants influence the well-known regional differences in incidences, since we can identify environmental risk factors that may either initiate the autoimmune process or promote already ongoing beta cell damage in different countries. For this, population-based epidemiological studies are necessary to identify risk determinants that may be useful for primary prevention strategies. PMID- 17130530 TI - Association of SUMO4, as a candidate gene for IDDM5, with susceptibility to type 1 diabetes in Asian populations. AB - Recent study demonstrated that M55V variant in SUMO4 at IDDM5 was associated with susceptibility to type 1 diabetes. Subsequent studies, however, showed inconsistency in the association. To clarify the population-wide effect on the association of SUMO4 with type 1 diabetes, we have performed meta-analysis including our own data in Asian populations, which confirmed a highly significant association in Asian populations (summary odds ratio [OR]: 1.29, P = 7.0 x 10( 6)), but indicated significant heterogeneity in the genetic effect of the SUMO4 gene on type 1 diabetes among diverse ethnic groups. These observations indicated the association of SUMO4 with type 1 diabetes in Asian populations. PMID- 17130531 TI - The gene for human IL-21 and genetic susceptibility to type 1 diabetes in the Japanese. AB - Type 1 diabetes is under polygenic control both in humans and the NOD mouse. Recently a possible role of IL-21 in the pathogenesis of type 1 diabetes was demonstrated in the NOD mouse. Furthermore, the murine IL-21 gene is mapped to the Idd3 interval, making the human IL-21 gene (IL21) a functional as well as positional candidate for susceptibility. We therefore screened sequence variants of IL21 and studied the association with type 1 diabetes. Preliminary data showed no association of IL21 polymorphisms with the disease, suggesting that IL21 plays little role in susceptibility to type 1 diabetes in Japanese. PMID- 17130532 TI - Genetics of type 1 diabetes: similarities and differences between Asian and Caucasian populations. AB - Transracial studies are a powerful tool for genetic association studies of multifactorial diseases, such as type 1 diabetes. We therefore studied the association of candidate genes, HLA, INS, CTLA4, PTPN22, and SUMO4, with type 1 diabetes in Asian populations in comparison with Caucasian populations. Class II HLA was strongly associated with type 1 diabetes in both Asian and Caucasian populations, but alleles associated with type 1 diabetes are different among different ethnic groups due to difference in allele distribution in general populations. INS was associated with type 1 diabetes in both Japanese and Caucasian populations, but frequency of disease-associated haplotype was markedly higher in Japanese than in Caucasian populations. CTLA4 association was reported for both type 1 diabetes and autoimmune thyroid diseases (AITD) in Caucasian populations, but the association with type 1 diabetes was concentrated in a subset of patients with AITD in Japanese. A variant (R620W) of PTPN22 was consistently associated with type 1 diabetes in Caucasian populations, but the variant was absent in Asian populations including Japanese. M55V variant of SUMO4 was significantly associated with type 1 diabetes in Asians, but genetic heterogeneity between Asian and Caucasian populations was suggested. These data indicate the importance of transracial studies with a large number of samples in each ethnic group in genetic dissection of type 1 diabetes. PMID- 17130533 TI - Immunopathological and genetic features in slowly progressive insulin-dependent diabetes mellitus and latent autoimmune diabetes in adults. AB - In 1982 we proposed the presence of a subtype of type 1 diabetes [slowly progressive insulin-dependent diabetes mellitus (SPIDDM)], which was characterized by persistently positive islet cell antibody, late age of onset, noninsulin-dependent diabetes, and slowly progressive beta cell failure. Since then many studies demonstrated that this subtype of type 1 diabetes is prevalent in many ethnic groups and was later called the latent autoimmune diabetes in adults (LADA). Recent epidemiological studies reported that about 10% of patients with apparent type 2 diabetes have at least one autoantibodies against islet specific antigen with high potential to progress to insulin-dependent state. Between SPIDDM and LADA some differences are reported in terms of some genetic predispositions including HLA class II and class I genes, vitamin D receptor gene, and CTLA4 genes. Common features in SPIDDM and LADA including preserved beta cells at the onset of diabetes and weak T cell response to residual beta cells suggest that these subtypes of type 1 diabetes are suitable candidates for prevention treatment for further progression of beta cell failure. PMID- 17130534 TI - Genes influencing innate and acquired immunity in type 1 diabetes and latent autoimmune diabetes in adults. AB - DQ8 and DQ2 are associated with susceptibility to and DQ6 with protection from type 1 diabetes mellitus (T1DM). A set of polymorphic genes, called MHC class I chain-related genes (MIC-A) in HLA class I region interact with NK cells. In Italians, MICA allele 5 increases T1DM risk by 6.1. Together with HLA-DQ8 and DQ2 the risk increases severalfold. HLA class I genes, also identified as susceptibility genes for T1DM, interact with polymorphic killer immunoglobulin like receptors (KIR) on NK cells. HLA-DQ8 and DQ2 and MICA-5 in Swedish and other populations also show positive association with disease. Studies on KIR in Latvian patients with T1DM also suggest a role for KIR in the etiology of T1DM. The results from MICA and KIR studies suggest that polymorphism of these genes of the innate immune system identify possible defects in the first line of antiviral defense in the etiology of T1DM. Screening for these genes could be important in the prediction strategies for T1DM. PMID- 17130535 TI - Progression of autoimmune diabetes: slowly progressive insulin-dependent diabetes mellitus or latent autoimmune diabetes of adult. AB - Autoimmune diabetes is due to destruction of insulin-secreting beta islet cells by an immune-mediated process, which is induced and promoted by the interaction of genetic and environmental factors. This form of diabetes is one of a group of autoimmune diseases that affect about 10% of the population in the developed world. The detection of diabetes-associated autoantibodies, including glutamic acid decarboxylase antibodies (GADA), islet cell antibodies (ICA), and insulinoma associated (IA-2) autoantibodies is widely held to reflect an underlying autoimmune pathology but the clinical features associated with the presence of these diabetes-associated autoantibodies is highly variable ranging from lack of symptoms with normal glucose tolerance to catastrophic and potentially fatal diabetic ketoacidosis. It is the purpose of this article to establish the range of metabolic features associated with diabetes-associated autoimmune changes and discuss how this metabolic spectrum itself reflects a spectrum of immune and clinical changes that cast light on the nature of autoimmune diabetes. PMID- 17130536 TI - Immunomodulation for the prevention of SPIDDM and LADA. AB - Type 1 diabetes may occur at any age, in young individuals before or after adolescence, during middle age life, or even in the elderly. When diagnosed in adults it is characterized by the presence of islet cell-related autoantibodies (ICA), in particular GAD and IA2 (less common) and very rarely insulin autoantibodies (IAA). Baseline C-peptide at diagnosis of type 1 diabetes can identify different patient populations according to when the disease is diagnosed depending on age. A key question is whether the process of beta cell destruction follows the same pattern in patients diagnosed in young age, soon after adolescence, or in adult age. The terms SPIDDM--slowly progressive insulin dependent diabetes mellitus, and LADA--latent autoimmune diabetes in adults, have been considered synonymous on most grounds based on the fact that with this form of diabetes we intend a form of diabetes that has an autoimmune basis that eventually will require insulin for its treatment sometime after diagnosis. Therapeutic approaches are similar for prevention and treatment of SPIDDM or LADA, including both specific and nonspecific immunomodulation. For specific immunomodulation the attention is focused on DiaPep277, GAD, and insulin, and for nonspecific immunomodulation on 1,25 dihydroxy-vitamin D3 (calcitriol) and thiazolidinediones. Current trials in SPIDDM/LADA with both specific and nonspecific immunomodulation seem promising. Response to therapy varies according to age and residual beta cell function at diagnosis of SPIDMM/LADA. Results in beta cell protection with different agents can also help to identify differences, if any, between SPIDMM and LADA. PMID- 17130537 TI - Heightened interferon-alpha/beta response causes myeloid cell dysfunction and promotes T1D pathogenesis in NOD mice. AB - Increasing attention is drawn to the contributions of abnormalities in both innate and acquired immune responses to the pathogenesis of autoimmune diseases, such as type 1 diabetes (T1D). Dendritic cells (DC) are critical immune cells linking innate and acquired immune responses and previous studies in NOD mice suggest abnormalities in these cells. To address DC dysregulation we examined kinetic global gene expression in NOD and B6 GM-CSF/IL-4-induced bone marrow derived DC following lipopolysaccharide (LPS)-stimulation. We identified expression differences in over 300 genes including a cluster of 16 interferon (IFN-alpha/beta) target genes overexpressed in NOD DC. Mechanistically, heightened IFN-alpha/beta responses were not due to increased production of this cytokine, IFN-gamma priming or increased Syk kinase activity. We found, however, heightened responses to IFN-alpha/beta in NOD versus B6 as demonstrated by increased type 1 IFN target gene expression, for example, IRF-7, in NOD DC and macrophages. Analysis of multiple congenic strains demonstrated that the Idd5 susceptibility region largely governed heightened IFN-alpha responses. Of interest, heightened IFN-alpha/beta response in NOD mice was not confined to hematopoietic cells but was also seen in the pancreas and beta cells. Compounding the IFN-alpha response defect, NOD mice harbor significantly more PDC in spleen in comparison to B6 and produce four- to sixfold more IFN-alpha when stimulated with CpG. Finally, treatment of NOD mice with IFN-alpha inducing agents, for example, high-dose poly I:C accelerates diabetes in both female and male mice. The abnormalities in the IFN-alpha/beta axis appear to play a significant role in T1D pathogenesis. PMID- 17130538 TI - Immunolocalization of monocyte chemoattractant protein-1 in islets of NOD mice during cyclophosphamide administration. AB - The molecular processes that initiate insulitis in type 1 diabetes remain unclear. Chemokines, such as monocyte chemoattractant protein-1 (MCP-1), mediate chemotaxis and leukocyte migration to inflammatory sites. Although MCP-1 mRNA has been shown in islets isolated from NOD mice at 2 weeks of age and at later stages, the cellular sources of this chemokine, at the protein level, and its role in insulitis are unclear. The aims of the present study were to employ immunohistochemical techniques to examine the expression of MCP-1 and quantify its cellular sources in islets of NOD mice both after cyclophosphamide (Cy) administration and in spontaneous diabetes. Tissues were examined at days 1 (=day 73, first day of Cy), 4, 7, 11, and 14 and in age-matched control NOD mice. Pancreatic sections from NOD mice without Cy administration were also studied between days 21-65 and at onset of diabetes and from adult CD-1 mice. In the Cy group, a small number of peri-islet macrophages were immunopositive for MCP-1 at day 1 whereas at day 4, the number declined but increased subsequently at day 7. In the same group, it increased markedly at days 11 and 14 compared with age matched control NOD mice. In young NOD mice, MCP-1 was present in selective macrophages in islets with early insulitis (day 45) but was absent at diabetes onset. MCP-1 was undetectable in beta cells and in most T cells. Islets from adult CD-1 mice did not show immunostaining for MCP-1. We conclude that MCP-1 is expressed in a proportion of islet and exocrine macrophages. This expression increases during the later stages of Cy-induced diabetes. Thus, MCP-1 positive macrophages that migrate to the islet periphery during the early stages of Cy induced diabetes and preceding spontaneous diabetes may augment insulitis by further attracting macrophages and T cells. PMID- 17130539 TI - Young NOD mice show increased diabetes sensitivity to low doses of streptozotocin. AB - In type 1 diabetes, environmentally induced early-limited beta cell damage may pre-empt the subsequent immune-mediated beta cell destruction. Low doses of streptozotocin (Stz), given early to diabetes-prone mice, may cause limited beta cell destruction during the early phase and precipitate diabetes. Here, we aimed to see if young NOD mice are more diabetes-sensitive to various multiple low doses of Stz than non-diabetes-prone mice. We also determined the molecular pathology of islets following administration of the diabetogen. Female NOD and CD 1 mice received 5 daily doses of Stz at day 21 (20, 30, and 40 mg/kg body weight; 18 mice per group) or diluent, and diabetes was monitored. Pancreas were studied histochemically and immunohistochemically at various time points after Stz administration. Following administration of Stz, NOD mice showed a much earlier onset and increased diabetes rate, at all three doses, than CD-1 mice. By day 80, the final diabetes rates following the 40, 30, and 20 mg dose in NOD mice were 95%, 85%, and 33%, respectively, compared with 33%, 28%, and 5.5%, respectively, in CD-1 mice. However, following the 20 mg dose, only 2 of the 12 remaining NOD mice developed the disease between 90 and 250 days compared with 19 of 24 NOD mice that did not receive Stz at day 21. Stz-administered NOD and CD-1 mice showed an initial loss of beta cells, with redistribution of islet endocrine cells, early macrophage infiltration, and increasing insulitis. PMID- 17130540 TI - Contribution of class III MHC to susceptibility to type 1 diabetes in the NOD mouse. AB - A recombinant major histocompatibility complex (MHC) with the same class III region as the NOD mouse, but different class II region from the NOD mouse was identified in the NON mouse, and NOD mice congenic for this recombinant MHC, NOD.NON-H2, was established. None of the congenic mice homozygous for the NON MHC developed type 1 diabetes, indicating that the NOD MHC is necessary for the development of type 1 diabetes. A small portion of MHC heterozygotes developed late-onset type 1 diabetes, suggesting the contribution of class III MHC to type 1 diabetes susceptibility. PMID- 17130541 TI - MHC-linked susceptibility to type 1 diabetes in the NOD mouse: further localization of Idd16 by subcongenic analysis. AB - Although major histocompatibility complex (MHC)-linked susceptibility is the strongest component, recent studies demonstrated that MHC-linked susceptibility to type 1 diabetes consists of multiple components both in humans and non-obese diabetic (NOD) mouse. In the NOD mouse, Idd16 has been mapped to the region adjacent to, but distinct from Idd1 in the MHC class II region. Establishment of subcongenic NOD.CTS-H2 lines that possess the same MHC class II as the NOD mouse but non-NOD-derived chromosomal region in its adjacent regions, would facilitate further narrowing down of the localization of Idd16. PMID- 17130542 TI - Long-term prevention of diabetes and marked suppression of insulin autoantibodies and insulitis in mice lacking native insulin B9-23 sequence. AB - We analyzed double native insulin gene knockout NOD mice with a mutated (B16:alanine) proinsulin transgene at multiple ages for the development of insulin autoantibodies, insulitis, and diabetes. In contrast to mice with at least one copy of a native insulin gene that expressed insulin antibodies, only 2 out of 21 (10%) double native insulin gene knockout mice with a mutated insulin transgene developed insulin autoantibodies. Of 21 double insulin knockout mice sacrificed between 10 to 48 weeks of age, only 5 showed minimal insulitis versus 100% of wild-type NOD and more than 90% of insulin 1 knockout mice. Consistent with robust suppression of insulin autoantibodies and insulitis, no double insulin knockout mice developed diabetes. In that the B9-23 peptide with B16A is an altered peptide ligand inducing Th2 responses, we analyzed transfer of splenocytes into NOD.SCID mice. There was no evidence for regulatory T cells able to inhibit transfer of diabetes by diabetogenic NOD splenocytes. Insulin peptide B9-23 is likely a crucial target for initiation of islet autoimmunity and further mutation of the sequence will be tested to attempt to eliminate all anti-islet autoimmunity. PMID- 17130543 TI - Eicosanoid imbalance in the NOD mouse is related to a dysregulation in soluble epoxide hydrolase and 15-PGDH expression. AB - Eicosanoids promote or resolve inflammation depending on the class produced. Macrophage from nonobese diabetic (NOD) mouse produce increased proinflammatory lipid mediators and low levels of antiinflammatory lipoxin A4 (LXA4). The enhanced proinflammatory eicosanoids is secondary to increased cyclooxygenase-2 (Cox-2) expression and low levels of prostaglandin/leukotriene catabolic enzyme, 15-hydroxyprostaglandin dehydrogenase (15-PGDH). Deficient LXA4 production is not due to deficient lipoxygenase (LO) activity, but is related to increased soluble epoxide hydrolase (sEH), involved in metabolism of anti-inflammatory epoxyeicosatrienoic acids (EET). These aberrations in eicosanoid biology suggest that inflammation in the NOD mouse is likely to be prolonged and robust and may contribute to type 1 diabetes (T1D) pathogenesis. PMID- 17130544 TI - Pancreatic autoimmunity induction with insulin B:9-23 peptide and viral mimics in the NZB mouse. AB - To create a new experimental model of diabetes, we used the New Zealand Black (NZB) mouse as a potential model. NZB mice were immunized with B:9-23 insulin peptide in IFA and the viral mimic, poly(A:U). No diabetes was observed but blood glucose was significantly higher in the B:9-23 peptide group compared to controls. Insulin autoantibodies (IAA) were only induced in groups given the B:9 23 peptide. B:9-23 alone induced peri-insulitis. We demonstrate insulin autoimmunity in the NZB mouse using the insulin peptide B:9-23 and viral mimics. The reason for the protection from diabetes despite the presence of autoimmunity is currently not established. PMID- 17130545 TI - Viruses cause type 1 diabetes in animals. AB - More than 10 viruses have been reported to be associated with the development of type 1 diabetes-like symptoms in animals, with the best evidence coming from studies on the D variant of encephalomyocarditis (EMC-D) virus in mice and Kilham rat virus (KRV) in rats. A high titer of EMC-D viral infection results in the development of diabetes within 3 days, primarily due to the rapid destruction of beta cells by viral replication within the cells. A low titer of EMC-D viral infection results in the recruitment of macrophages to the islets. Soluble mediators produced by activated macrophages play a critical role in the destruction of residual beta cells. A single amino acid at position 776 of the EMC viral genome controls the diabetogenicity of the virus. In contrast, KRV causes autoimmune type 1 diabetes in diabetes-resistant BioBreeding (DR-BB) rats without direct infection of beta cells. Macrophages play an important role in the development of diabetes in KRV-infected DR-BB rats. As well, KRV infection preferentially activates effector T cells, such as Th1-like CD45RC(+)CD4(+) T cells and CD8(+) T cells, and downregulates regulatory T cells, such as Th2-like CD45RC(-)CD4(+) T cells. This results in the breakdown of the immune balance, contributing to the development of diabetes in KRV-infected DR-BB rats. PMID- 17130546 TI - Characterization of PAF-AH Ib1 in NOD mice: PAF-AH may not be a candidate gene of the diabetes susceptibility Idd4.1 locus. AB - We recently mapped Idd4 to a 5.2 cM interval on chromosome 11 with two subloci, Idd4.1 and Idd4.2, in nonobese diabetic (NOD) mice. Based on the localization of platelet-activating factor acetylhydrolase Ib1 (PAF-AHIb1) and the decreased activity of PAF-AH in type 1 diabetes (T1D) patients, we hypothesized that PAF AHIb1 in Idd4.1 is a candidate gene. The PAF-AHIb1 gene in NOD mice was cloned and sequenced, and its expression and function were studied. No polymorphisms were detected in PAF-AHIb1 cDNA between NOD and B6 mice. The expression of PAF-AH Ib1 at the mRNA and protein levels was found to be similar in different tissues between NOD and B6 mice. PAF-AH activity does not differ in the pancreatic islets or spleen between NOD and B6 mice. Our findings suggest that PAF-AH Ib1 may not be a diabetes-susceptibility gene in the Idd4.1 sublocus. PMID- 17130547 TI - Peptide-pulsed immature dendritic cells reduce response to beta cell target antigens and protect NOD recipients from type I diabetes. AB - Our previous work demonstrated peptide-pulsed mature myeloid dendritic cells (DC) presenting beta cell antigens induce tolerance. Here we determine whether immature DC (iDC) presenting dominant (insulin beta9-23 chain, proinsulin C19-A3) or ignored (glutamic acid decarboxylase 65(78-97)) antigen determinants promote tolerance. Nonobese diabetic (NOD) mice were given injections of either unpulsed or peptide-pulsed myeloid iDC beginning at 9 weeks of age for 3 consecutive weeks. Diabetes incidence in recipients of unpulsed iDC was comparable to unmanipulated animals ( approximately 80%), whereas GAD65(78-97) pulsed iDC recipients were protected from the disease (P = 0.05). We also analyzed splenic T cell proliferation responses to the panel of studied peptides in diabetic and nondiabetic recipients. When stimulated with insulin or proinsulin peptide, nondiabetic mice receiving the peptide-pulsed iDC had a 21- to 31-fold or 3.9- to 9.0-fold reduction in T cell response, respectively, as compared to the response of diabetic unpulsed recipients. However, only a 2.6- to 3.1-fold reduction in response to beta chain peptide, and a 1.5- to 3.4-fold reduction in proinsulin response were observed in diabetic mice receiving peptide-pulsed iDC. The reduction was not specific to the immunizing peptide, as reduced proliferation was observed to other diabetes-target peptides. We conclude that protective iDC based therapies require target antigen presentation, and ignored determinants may be preferable perhaps due to an available naive T cell repertoire. In addition, iDC presenting peptides induce a nonspecific reduction in T cell responses to beta cell antigens, possibly through the induction of regulatory T cells. PMID- 17130548 TI - New members of the interleukin-12 family of cytokines: IL-23 and IL-27 modulate autoimmune diabetes. AB - Multiple low doses of streptozotocin (5 x 40 mg/kg) given to susceptible male C57BL6 mice induced delayed and sustained hyperglycemia accompanied by body weight loss, mononuclear cell infiltration in the islet, and apoptosis of beta cells. Shorter regimes (4 x 40 mg/kg) did not have such effect. Administration of IL-23 at a dose of 400 ng/mL for 10 consecutive days concomitantly with this subdiabetogenic regimen of STZ, however, induced significant hyperglycemia, weight loss, and mononuclear cellular infiltration. The same regimen of IL-27 induced milder effect on glycemia and no weight loss inspite of a massive peri islet and intra-islet infiltration of mononuclear cells. The molecular mechanisms underlying the actions of these cytokines on diabetogenesis is under study. PMID- 17130549 TI - Glutamic acid decarboxylase-specific CD4+ regulatory T cells. AB - It is known that CD4(+) regulatory T cells (Tr cells) play a central role in inducing immune tolerance in animals and humans. Compared to polyclonal Tr cells, autoantigen-specific Tr cells are more potent at blocking pathogenic immune responses. In order to better understand the role of Tr cells in controlling type 1 diabetes development and to help design effective antigen-specific cell-based therapeutic methods to treat the disease, it is necessary to: (a) determine the antigen specificity of Tr cells; (b) study how antigen-specific Tr cells behave in vivo; (c) investigate the interaction of Tr cells with pathogenic T cells (Tpath cells) and determine whether such interaction correlates with the progression or inhibition of diabetes; and (d) determine the cellular and molecular mechanisms underlying the regulation of diabetes by Tr cells. We have addressed these questions with a focus on the studies of glutamic acid decarboxylase (GAD)-specific T cells. Previous studies have suggested that GAD specific T cells play a key role in type 1 diabetes. Treatment of NOD mice with GAD or its peptides can prevent the progression toward overt disease. The preventive effect could be due to either the deletion of antigen-specific pathogenic T cells or the induction of potent antigen-specific Tr cells. Using antigen-specific I-Ag7 tetramers we have isolated several populations of GAD peptide-specific T cells from diabetes-prone NOD and diabetes-resistant NOR mice. Herein, we summarize our studies on the role of these GAD peptide-specific T cells in type 1 diabetes. We present evidence that supports the hypothesis that the repertoire of T cells specific for these GAD peptides is biased toward Tr cells that inhibit diabetes rather than toward pathogenic T cells that induce diabetes. PMID- 17130550 TI - Metabolism genes are among the differentially expressed ones observed in lymphomononuclear cells of recently diagnosed type 1 diabetes mellitus patients. AB - The large-scale differential gene expression in lymphomononuclear cells of six patients with recently diagnosed type), and six normal individuals matched to patients for sex and age were studied. Glass slides containing 4608 cDNAs from the IMAGE library were spotted using robotic technology. Statistical analysis was carried out by the SAM program, and gene function assessed by the FATIGO program. Thirty differentially expressed genes (21 induced and 9 repressed) were disclosed when DM-1 patients were compared with controls. Although presenting with distinct biological function, most of the induced or repressed genes were related with protein, phosphate, DNA, RNA, carboxylic acid, and fatty acid metabolism. Although some of these genes have been previously associated with the pathogenesis of T1DM, many other genes were identified for further studies. PMID- 17130551 TI - In vitro TNF-alpha and IL-6 production by adherent peripheral blood mononuclear cells obtained from type 1 and type 2 diabetic patients evaluated according to the metabolic control. AB - Tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) levels were evaluated in lipopolysaccharide (LPS)-stimulated cell cultured monocytes obtained from 24 type 1 and type 2 diabetic patients presenting inadequate (IN) or adequate (AD) metabolic control, and in 21 healthy individuals paired to patients for sex and age. The TNF-alpha levels in stimulated cultures of diabetic patients were similar to healthy individuals, and type 1 diabetic patients showed increased IL-6 supernatant levels. The tendency toward increased TNF-alpha and IL 6 levels was observed with metabolic control of type 1 and type 2 diabetic patients, suggesting that the control of diabetes improves the capacity of activation and maintenance of the proinflammatory immune response. PMID- 17130552 TI - T cell immunity to glutamic acid decarboxylase in fulminant type 1 diabetes without significant elevation of serum amylase. AB - We encountered three patients with fulminant type 1 diabetes whose serum amylase levels were not elevated and evaluated their immunological characteristics. Although all three patients had no antibodies to islet antigens including glutamic acid decarboxylase (GAD), GAD-reactive T lymphocytes were detected in two patients. Combined with the findings that human leukocyte antigen (HLA) class II haplotype associated with fulminant type 1 diabetes is the same as that of autoimmune type 1 diabetes, an immune process similar to autoimmune type 1 diabetes may be involved in at least a part of fulminant type 1 diabetes. PMID- 17130553 TI - Expression levels of CXC chemokine receptors 3 are associated with clinical phenotype of type 1 diabetes. AB - Type 1 diabetes is recognized as one of T helper 1 cell (Th1)-mediated diseases. The purpose of this article was to investigate the expression levels of CXC chemokine receptor 3 (CXCR3) and CC chemokine receptor 5 (CCR5) on CD4 T cells as Th1 markers in Japanese patients with type 1 diabetes and control subjects. A total of 72 patients with type 1 diabetes and 24 healthy subjects were enrolled. Their peripheral mononuclear cells were obtained and stained with anti-CXCR3, anti-CCR5, and anti-CD4 monoclonal antibodies. Flow-cytometric analysis was performed and patients were classified according to their onset pattern as fulminant, typical, or slow onset. Statistical analysis was performed using ANOVA. CXCR3 expression on CD4 T cells in patients with a fulminant pattern of onset was significantly lower than that in the other groups, and that in patients with a typical pattern of onset was significantly higher than that in the other groups. CCR5 expression on CD4 T cells was not different among the three clinical phenotypes. CXCR3 expression level is associated with the onset pattern of type 1 diabetes. Further studies are needed to clarify the role of chemokines in type 1 diabetes. PMID- 17130554 TI - HLA class I epitope discovery in type 1 diabetes. AB - Type 1 diabetes mellitus (T1DM) results from the destruction of beta cells by autoantigen-specific T cells. In the non-obese diabetic (NOD) mouse model, CD8+ T cells play an essential role in both the initial triggering of insulitis and its destructive phase, and proinsulin (PI) is one of the dominant target antigens (Ags). However, little is known about the beta cell epitopes presented by HLA class I molecules and recognized by human CD8+ T cells. We and other groups recently applied reverse immunology approaches to identify HLA class I-restricted PI epitopes. To establish an inventory of potential naturally processed epitopes, whole human PI or the transitional region between the B-chain and C-peptide were digested with purified proteasome complexes. By combining proteasome digestion data with epitope prediction algorithms, candidate epitopes restricted by HLA A2.1 and other HLA class I molecules were identified. We validated immunogenicity and natural processing of the identified PI epitopes in HLA-A2.1-transgenic mice, while others demonstrated recognition of multiple PI epitopes by CD8+ T cells from T1DM and healthy subjects in the context of different HLA class I molecules. These results demonstrate the power of reverse immunology strategies for epitope discovery. DNA vaccination of HLA-transgenic mice may be another rapid and efficient reverse immunology approach to map additional epitopes derived from other T1DM Ags, such as IA-2 and glutamic acid decarboxylase 65 (GAD 65). Transfer of this information to Elispot- and MHC tetramer-based assay formats should allow to reliably detect and characterize autoreactive CD8+ T cell responses in T1DM, and may open new avenues for early T1DM diagnosis and immune intervention. PMID- 17130555 TI - Upregulation of Foxp3 expression in mouse and human Treg is IL-2/STAT5 dependent: implications for the NOD STAT5B mutation in diabetes pathogenesis. AB - Regulatory T cells (Treg), characterized as CD4(+)/CD25(+hi) T cells, are critical for sustaining and promoting immune tolerance. Treg are highly dependent on IL-2 and IL-2 signaling to maintain their numbers and function and interruption of this pathway promotes autoimmunity. The transcription factor, Foxp3, is also required for Treg function as defective Foxp3 promotes autoimmunity in both mice and humans. We previously reported a point mutation in the DNA-binding domain of the NOD STAT5B gene that limits DNA binding when compared to wild-type STAT5 mice. Based on the presence of five STAT5B consensus sequences in the Foxp3 promotor, we hypothesized a critical linkage between IL-2 signaling/STAT5B and Foxp3 expression in Treg. Our data show IL-2 activates long form (LF) STAT5 and sustains Foxp3 expression in Treg. In contrast, CD4(+)/CD25( ) T cells do not active LF STAT5 and do not express Foxp3 under the same conditions. In addition, blocking LF STAT5 activation with a Jak inhibitor (AG 490) significantly reduced Foxp3 expression in Treg. Examination of human Treg using flow cytometry and intracellular staining for Foxp3 expression likewise demonstrates that IL-2 maintains Foxp3 expression through LF STAT5 signaling. These studies reveal a critical link between IL-2 mediated JAK-STAT5 signaling and the maintenance of Foxp3 expression in Treg of mice and humans. PMID- 17130556 TI - Normal T cell development in the absence of thymic insulin expression. AB - Ectopic expression of insulin in thymus has been suggested to be involved in tolerance induction against pancreatic beta cells and in type 1 diabetes (T1D) pathogenesis. However, it is not known whether thymic insulin expression would also influence thymocyte maturation and differentiation. To address these questions, we have used mice that are insulin deficient. Early fetal thymi were cultured in fetal thymic organ cultures (FTOCs) and the development of thymocytes was studied by flow cytometry. The results revealed no significant difference in thymocyte maturation in the absence of thymic insulin. Taken together, these data do not support a role for thymic insulin in thymocyte differentiation and growth. PMID- 17130557 TI - Antigenic determinants to GAD autoantibodies in patients with type 1 diabetes with and without autoimmune thyroid disease. AB - Type 1 diabetes (T1D) is frequently associated with other autoimmune diseases. Most T1D patients' sera contain two distinct glutamic acid decarboxylase (GAD) antibody specificities, of which one targets an epitope region in the middle third of GAD65 (amino acids 221-359) and the other targets the carboxy-third of GAD65 (amino acids 453-569). Using five chimeric GAD65/GAD67 proteins to maintain conformation-dependent epitopes of GAD65, we compared the humoral repertoire of antibodies from 127 T1D patients with and without autoimmune thyroid diseases (ATD). Thirty-one patients with T1D (24%) expressed antithyroid autoantibodies ATA and 22 patients (17%) had ATD in comparison to 6% of age-matched controls having ATA. GAD65-antibody-positive patients much more often (28% versus 5%, P < 0.0004) had ATD. Of 66 GAD65-autoantibody-positive T1D patients, 34 had autoantibodies reacting with both middle and carboxy epitopes. Autoantibodies of the other 32 reacted with middle, carboxy, or other epitopes but not with both middle- and carboxy-third. Those with GAD65 autoantibodies reacting with both middle- and carboxy-third had less ATD. Of 22 (23%) patients with ATD, 5 compared to 29 of 47 (62%) T1D patients without ATD had GAD65 autoantibodies reacting with both middle- and carboxy-third (relative risk = 0.2, P < 0.01). These results indicate that there are both similarities and differences in the humoral response to GAD65 in ATD and T1D, and expression of antibodies to middle- and carboxy third at the same time is a feature specific to T1D. PMID- 17130558 TI - TSH receptor antibodies in subjects with type 1 diabetes mellitus. AB - The research was undertaken to study the prevalence of TSH receptor antibody positivity in patients with type 1 diabetes. A total of 74 subjects with type 1 diabetes were enrolled in this cross-sectional study. Thyroid function test and assessment of thyroid autoimmunity with anti-TPO and TSH receptor antibody were done in all patients. A total of 33 males and 41 females with type 1 diabetes were studied. The prevalence of TSH receptor antibody positivity alone was 18%. The prevalence of thyroid autoimmunity with anti-TPO as a marker was 28%; the prevalence increased to 43% when TSH receptor antibody was also measured. Majority of the subjects with antithyroid antibody positivity were also positive for GAD65 antibodies. As a significant proportion of type 1 diabetic subjects have positivity to TSH receptor antibody, we suggest that larger studies should be conducted to study the benefits of TSH receptor antibody-based screening for thyroid dysfunction in type 1 diabetic subjects. As the TSH receptor antibodies could be of the stimulating or of the blocking type, subjects with antibody positivity could be at risk of developing hyperthyroidism or hypothyroidism. PMID- 17130559 TI - Time-resolved fluorescence imaging of islet cell autoantibodies. AB - Time-resolved fluorescence of lanthanide chelates has been widely used in bioanalytical assays. Long fluorescence time, large Stokes shift, and minute fading out of the fluorescence over years are major advantages of the lanthanides over the conventional fluorescent dyes. We have now applied time-resolved fluorescence imaging (TRFI) also for measurement of type 1 diabetes mellitus (T1DM)-related islet cell autoantibodies (ICA). Retaining the accuracy of conventional ICA, TRFI has over 10 times better signal-to-noise ratio than the conventional fluorochromes. The technology allows objective determination of fluorescence intensity with the camera and computer software, and serial dilutions for obtaining the antibody titer in autoantibody-positive samples are unnecessary. We now describe the TRFI as a method and its application for measurement of ICA. PMID- 17130560 TI - MHC class I chain-related gene-A is associated with IA2 and IAA but not GAD in Swedish type 1 diabetes mellitus. AB - In type 1 diabetes mellitus (T1DM), the frequency of antibodies against insulin (IAA), glutamic acid decarboxylase-65 (GAD65), ICA512/IA2 (IA2), and islet cell antigens (ICA) vary with human leukocyte antigen (HLA) composition of the patient. IAA, IA2 autoantibodies, and ICA are increased in DQ8 positives; GAD65 antibodies are increased in DQ2 positives. MHC class I chain-related gene-A (MICA) is another genetic marker that has been proposed to be associated with T1DM. In this article, we looked at microsatellite polymorphism of MICA and its association with autoantibodies (IAA, IA2, and GAD65) in Swedish T1DM patients and if the association explains its importance in early events in autoimmune response. We studied 635 T1DM patients between 0-35 years. Frequency of MICA5/5 was positively associated with the formation of IAA and IA2 antibodies considered individually or in combination (odds ratio [OR], 95% CI, Pc: [IAA+ versus IAA-]: 4.94, 2.09-11.62, <0.0005; [IA2+ versus IA2-]: 2.65, 1.52-4.59, 0.0015; [IAA and/or IA2+ versus rest]: 9.83, 2.37-40.78, <0.0015; [IAA and IA2+ versus rest]: 3.51, 2.01-6.15, <0.0015). Also, -5.1/5.1 was increased in IAA+ patients compared to IAA- patients (2.82, 1.64-4.83, <0.0005). All patients positive for -5/5 developed at least one of the three antibodies. Frequency of MICA5.1 was decreased in IAA+ (0.54, 0.36-0.81, 0.017), in IA2A+ (0.63, 0.45-0.88, 0.04), in IAA and/or IA2A+ (0.52, 0.33-0.84, 0.044), and in IAA and IA2A+ (0.55, 0.39-0.78, 0.0055) patients when compared with patients negative for corresponding antibodies. Frequency of MICA9, 5/5.1, and 5.1/9 was decreased in IAA+ compared to IAA- patients (0.51, 0.32-0.79, 0.021; 0.22, 0.11-0.44, <0.005; and 0.39, 0.22 0.69, 0.026, respectively). Frequency of MICA9 and -5.1/9 was also decreased in IAA and/or IA2 antibody-positive patients while MICA5/5.1 decreased in patients positive for IAA and IA2 antibody both together. IAA and IA2 antibodies are believed to appear early during the autoimmune reaction against beta cells. Thus, according to our data, MICA-5/5 and -5.1/5.1 is associated with early autoimmunity in T1DM patients. Our study suggests that MICA gene polymorphism is associated with autoantibody formation and that the polymorphism especially MICA5/5 and -5.1/5.1 are important in early events of autoimmune reaction. PMID- 17130561 TI - Predominance of the group A killer Ig-like receptor haplotypes in Korean patients with T1D. AB - Type 1 diabetes (T1D) is a T cell-mediated autoimmune disease in which pancreatic beta cells are selectively destroyed. Although autoimmune diseases are driven by inappropriate adaptive immunity, innate immunity may play a role in the development of T1D. To study the potential involvement of innate immunity in the pathogenesis of autoimmune disease, we investigated associations of the genes for 14 different killer Ig-like receptors (KIRs), the well-characterized receptors in natural killer cells, with Korean T1D patients. Genetic association analyses revealed that some of the KIR genes were associated with T1D. KIR2DL5 and 2DS2 genes were present at significantly low frequency in Korean T1D patients (P < 10( 4)). We did not detect any influence of ligand distribution on KIR association. With the haplotype assignments, 53% of the KIR haplotypes in the control are of type A. Compared with the control (P < 10(-3)) and autoantibody-negative patients (P < 10(-2)), the group A haplotype predominates in Korean patients with T1D. The KIR gene is associated with T1D and distribution differences between T1D and controls were not influenced by the HLA genes (DR-DQ-A-C). T1D, at least in Koreans, is associated with KIR genes, especially in the group A KIR haplotypes. There is a close relationship between innate and adaptive immunity. PMID- 17130562 TI - Frequency of CTLA-4 gene CT60 polymorphism may not be affected by vitamin D receptor gene Bsm I polymorphism or HLA DR9 in autoimmune-related type 1 diabetes in the Japanese. AB - One of the CTLA-4 SNPs, +6230G>A (CT60), has recently been reported to be related to susceptibility to type 1 diabetes and autoimmune thyroid disease. We have previously reported an association between acute-onset type 1 diabetes in Japanese and the Vitamin D receptor (VDR) gene Bsm I large B polymorphism, which is related to the Th1-type response. Moreover, we found a significant correlation between autoimmune-related type 1 diabetes with HLA DR9 and detection of GAD reactive Th1 (T helper 1)-type cells. In the present article, we tried to clarify whether the frequency of one of the CTLA-4 SNPs, +6230G>A (CT60), is affected by the VDR gene Bsm I polymorphism or by HLA DR9 in Japanese type 1 diabetics. The frequency of the CT60 GG genotype did not appear to be affected by either the VDR gene Bsm I large B polymorphism or HLA DR9. PMID- 17130563 TI - Genetic and functional evidence supporting SUMO4 as a type 1 diabetes susceptibility gene. AB - Genomewide linkage analyses since the early 1990s suggested over 20 genomic intervals that may contain susceptibility genes for type 1 diabetes. However, the identification of the specific genes in these intervals presents a formidable challenge due to a number of difficulties associated with genetic mapping and cloning of genes implicated in complex diseases. One of the difficulties is due to the presence of many weak and different susceptibility genes in different patients and populations, a phenomenon known as genetic heterogeneity. In 2004, we reported the cloning of a novel small ubiquitin-like modifier (SUMO) gene, SUMO4, in the IDDM5 interval on chromosome 6q25, and presented strong genetic and functional evidence suggesting that SUMO4 is a susceptibility gene for type 1 diabetes mellitus (T1DM). In this article, we will summarize genetic association data suggesting that SUMO4 is consistently associated with T1DM in the Asian populations while the association is more heterogeneous in the Caucasian populations. We will also discuss the possible molecular pathways through which sumoylation may regulate T1DM and autoimmunity. PMID- 17130564 TI - No association of TLR2 and TLR4 polymorphisms with type I diabetes mellitus in the Basque population. AB - The destruction of pancreatic beta cells that occurs in type 1 diabetes mellitus (T1DM) is mediated by the immune system, and evidence has accumulated supporting the implication of innate immune mediators. Toll-like receptors (TLRs) participate in the first line of immune defense through antigen recognition, and their ligands are mostly exogenous but can be host-derived as well. To test the possible role of TLRs in the development of T1DM, we studied different SNPs of TLR2 (N199N, S450S, R677W, and R753Q) and TLR4 (D299G, T399I, and S400N) in Basque families with T1DM. Several positions analyzed were not polymorphic in the Basque population. Genetic association analysis failed to demonstrate any association of these polymorphisms of TLR2 and TLR4 with T1DM in our population. The differences in TLR4 haplotype transmission to affected and unaffected offspring are indicative of a possible implication of TLR4 in disease risk but differences did not reach statistical significance. PMID- 17130565 TI - Association of SUMO4 M55V polymorphism with autoimmune diabetes in Latvian patients. AB - Small ubiquitin-related modifier (SUMO4), located in IDDM5, has been identified as a potential susceptibility gene for type 1 diabetes mellitus (T1DM). The novel polymorphism M55V, causing an amino acid change in the evolutionarily conserved met55 residue has been shown to activate the nuclear factor kappaB (NF-kappaB), hence the suspected role of SUMO4 in the pathogenicity of T1DM. The M55V polymorphism has been shown to be associated with susceptibility to T1DM in Asians, but not in Caucasians. Latent autoimmune diabetes in adults (LADA) is a slowly progressive form of T1DM and SUMO4 M55V has not been studied in LADA to date. The current study aims to test whether Latvians are similar to Caucasians in susceptibility to autoimmune diabetes (T1DM and LADA), with respect to SUMO4 M55V. We studied, age- and sex-matched, Latvian T1DM patients (n = 100) and healthy controls (n = 90) and LADA patients (n = 45) and healthy controls (n = 95). SUMO4 M55V polymorphism was analyzed using polymerase chain reaction (PCR) restriction fragment length polymorphism (RFLP). The allelic frequencies of the A and G alleles were compared with HLA DR3-DR4-positive and HLA DR3-DR4-negative patients to identify any potential relation between HLA DR3-DR4 and SUMO4 M55V. We found no significant association between SUMO4 M55V and T1DM susceptibility in Latvians, the results being in concurrence with the previous studies in Caucasians of British and Canadian origin. Comparison of the A and G alleles with HLA DR3-DR4 did not result in any significant P values. No significant association was found between SUMO4 M55V and LADA. SUMO4 M55V is not associated with susceptibility to T1DM and LADA in Latvians, and Latvians exhibit similarity to other Caucasians with respect to association of SUMO4 M55V with autoimmune diabetes. PMID- 17130566 TI - A second component of HLA-linked susceptibility to type 1 diabetes maps to class I region. AB - Type 1 diabetes is a polygenic disease with a major susceptibility locus, IDDM1, located in the human leukocyte antigen (HLA) region. Although class II loci, DR and DQ genes in particular, are major components of IDDM1, accumulating lines of evidence indicated that IDDM1 consists of multiple components and that non-class II genes in addition to class II genes contribute to susceptibility to and/or age at-onset of type 1 diabetes. To identify a second component of IDDM1, we investigated the association of a panel of polymorphisms in 2.2 Mb region of the HLA encompassing from class II to class I regions with type 1 diabetes. Polymorphisms types were: DRB1 and DQB1 in class II; two microsatellite markers, BAT2-GT and TNFa in class III; and, five microsatellite markers, STR-MICA, MIB, C1-3-1, C2-4-4, and C3-2-10 in class I region. A total of >200 Japanese patients and healthy control subjects were studied. Class II DRB1*0405 and DQB1*0401 were significantly associated with susceptibility to, but not with age-at-onset of, type 1 diabetes. C1-3-1, located near C locus, was significantly associated with not only susceptibility to, but also age-at-onset of type 1 diabetes. These data suggest that a second component of IDDM1 maps to the HLA class I region, contributing to susceptibility to as well as age-at-onset of type 1 diabetes. PMID- 17130567 TI - Molecular scanning of the gene for programmed cell death-1 (PDCD-1) as a candidate for type 1 diabetes susceptibility. AB - Multiple genes are involved in the susceptibility to autoimmune type 1 diabetes. The immunoreceptor programmed cell death-1 (PDCD-1), an inhibitory costimulatory molecule regulating peripheral tolerance, was reported to play a role in the development of type 1 diabetes, making the human PDCD-1 gene, PDCD1, as a candidate for disease susceptibility. In this article, we sequenced all 5 exons and exon-intron junctions of PDCD1 in Japanese subjects, and found 10 sequence variants. Preliminary data suggested no association of these polymorphisms with type 1 diabetes. These sequence variants are valuable for further studies to clarify contribution of PDCD1 to susceptibility to type 1 diabetes. PMID- 17130568 TI - Genetic determinants of type 1 diabetes across populations. AB - T1D results from autoimmune-mediated destruction of the pancreatic beta cells, a process that is conditioned by multiple genes and environmental factors. The main genetic determinants map to the major histocompatibility complex (MHC), and in particular DR and DQ, although, genes outside the MHC contribute, including the insulin gene, PTPN22, and CTLA-4. There are remarkable differences in genetic susceptibility to T1D between populations. We believe this variation reflects differing frequencies of diabetes causative and protective alleles and haplotypes, and thus remains a major genetic influence linked to the MHC region not accounted for by DR and DQ alleles. In this article, we discuss global variations in genetic susceptibility to T1D in view of current genetic understanding. PMID- 17130569 TI - TNFa-e microsatellite, HLA-DRB1 and -DQB1 alleles and haplotypes in Brazilian patients presenting recently diagnosed type 1 diabetes mellitus. AB - TNF microsatellite and HLA class II polymorphisms were studied in 28 recently diagnosed Brazilian patients presenting type 1 diabetes mellitus (T1DM) and in 120 healthy controls. TNFa-e and HLA-DRB1/DQB1 alleles were identified using sets of sequence-specific primers. Compared to controls, the DRB1*03 and DQB1*02 allele groups, TNFa1 allele, and the TNFa4-b5-c1-d4-e3 and TNFa10-b5-c1-d4-e3 haplotypes were overrepresented in patients. TNF microsatellite together with HLA polymorphisms is associated with type 1 diabetes in Brazilian patients, corroborating the participation of the MHC genes in disease susceptibility. PMID- 17130570 TI - Is HLA class II profile relevant for the study of large-scale differentially expressed genes in type 1 diabetes mellitus patients? AB - We have previously identified 30 differentially expressed genes when comparing recently diagnosed type 1 diabetes mellitus (DM-1) patients and controls paired for sex, age, and ethnic background. In this article we performed the hierarchical clustering of these genes taking into account the human-leukocyte antigen (HLA)-DRB1/DQB1 profile. The dendrogram obtained using the Cluster program grouped patients and controls into three clusters, one including individuals with no susceptibility alleles, another including individuals with at least three susceptibility alleles, and a third intermingling susceptibility/protective alleles. In addition to other variables, the results of the present article suggest that the major histocompatibility complex (MHC) class II profile may be of relevance for the study of a large-scale differentially expressed genes. PMID- 17130571 TI - Feasibility of a type 1 diabetes primary prevention trial using 2000 IU vitamin D3 in infants from the general population with increased HLA-associated risk. AB - Recent epidemiologic, immunologic, and NOD mouse studies suggest that intervention in the vitamin D system may be a successful method to prevent type 1 diabetes. Newborns at increased HLA-associated risk are randomized to receive either 400 or 2000 IU vitamin D3 by 1 month of age. We show that recruitment of babies from the general population for identification of HLA-associated risk status followed by enrollment to a randomized controlled prevention trial is feasible in Canada. PMID- 17130572 TI - Interleukin-10 plasmid construction and delivery for the prevention of type 1 diabetes. AB - Studies of animals with spontaneous autoimmune diabetes have revealed that autoreactive T cells that mediate islet beta cell destruction can be manipulated by the administration of Th(2) cytokines. In this article, the effect of interleukin-10 (IL-10) gene delivery was evaluated in vitro and in vivo with a novel IL-10 plasmid, pSI-IL-10-NFkappaB. In pSI-IL-10-NFkappaB, the expression of the IL-10 gene was driven by the SV40 promotor/enhancer. The nuclear factor kappaB (NFkappaB) binding sites were also introduced to facilitate nuclear transport of the plasmid in the cell. In vitro transfection assay with pSI-IL-10 NFkappaB showed a similar expression level of IL-10 to the plasmid without NFkappaB binding sites (pSI-IL-10). pSI-IL-10-NFkappaB and pSI-IL-10 were intravenously injected into 5-week-old nonobese diabetic (NOD) mice using polyethylenimine (PEI) as a gene carrier. Both groups had persistent gene expression, longer than 5 weeks, and secreted the similarly high IL-10 serum levels. Interestingly, the degree of insulitis in the pSI-IL-10-NFkappaB group was improved over the pSI-IL-10 group, PEI-only group, and noninjected controls. The serum glucose levels showed that single injection of pSI-IL-10-NFkappaB prevented the development of diabetes in 100% of the pSI-IL-10-NFkappaB-injected animals (5/5), while that of pSI-IL-10 prevented diabetes in 40% of the treated animals (2/5). These results suggest that pSI-IL-10-NFkappaB with PEI can effectively reduce the incidence of insulitis and type 1 diabetes in NOD mice. PMID- 17130573 TI - TEDDY--The Environmental Determinants of Diabetes in the Young: an observational clinical trial. AB - The aim of the TEDDY study is to identify infectious agents, dietary factors, or other environmental agents, including psychosocial factors, which may either trigger islet autoimmunity, type 1 diabetes mellitus (T1DM), or both. The study has two end points: (a) appearance of islet autoantibodies and (b) clinical diagnosis of T1DM. Six clinical centers screen newborns for high-risk HLA genotypes. As of December 2005 a total of 54,470 newborns have been screened. High-risk HLA genotypes among 53,560 general population (GP) infants were 2576 (4.8%) and among 910 newborns with a first-degree relative (FDR) were 194 (21%). A total of 1061 children have been enrolled. The initial enrollment results demonstrate the feasibility of this complex and demanding a prospective study. PMID- 17130574 TI - Protection from type 1 diabetes by vitamin D receptor haplotypes. AB - Vitamin D has been involved in the modulation of calcium and bone metabolism as well as in the immune system, where it suppresses the proliferation of activated T cells. These effects are exerted via the vitamin D receptor (VDR). Polymorphisms within this gene have been exhaustively studied in diverse autoimmune diseases but with inconsistent results. We previously reported a positive association of polymorphisms within the VDR gene (Apa I, Taq I, Bsm I, and Fok I). In the present article we extended our previous reports to seven additional polymorphisms (rs757343, rs9729, rs2853559, rs1989969, rs3847987, rs2238135, and rs4516035) in a larger set of German simplex type 1 diabetes families. Additionally we correlated serum levels of 25(OH)D(3) and 1,25(OH)(2)D(3) with VDR genotypes and haplotypes. The haplotypes "CG" (Taq I-Apa I), "CGG" (Taq I-Apa I-Tru I), "CGC" (Taq I-Apa I-Fok I), "GCTG" (rs9729-Taq I Apa I-Tru I), and "CGGC"(Taq I-Apa I, Tru I, Fok I) were less often transmitted, thus negatively associated with type 1 diabetes. Patients who carried the genotype "CC" of the rs3847987 polymorphism had higher median serum levels of 25(OH)D(3). Furthermore, the majority of patients with this genotype possessed normal serum levels of 25(OH)D(3). We conclude that variants of the VDR may confer a genetic protection from type 1 diabetes. Furthermore, normal serum levels of 25(OH)D(3) appear to correlate with a VDR genotype. This supports a role of vitamin D in the immune pathogenesis of type 1 diabetes. PMID- 17130575 TI - Living donor islet transplantation, the alternative approach to overcome the obstacles limiting transplant. AB - We performed the world's first successful living donor islet transplantation for unstable diabetes. A total of 408,114 islet equivalents were isolated from half a living pancreas and transplanted immediately to the recipient who was a 27-year old female. The donor was a 56-year-old female in good health, mother of the recipient. The islets functioned immediately, and the recipient was weaned completely from insulin on the 22nd posttransplant day, and has maintained excellent glycemic control since. The donor was discharged on the 18th postoperative day with normal oral glucose tolerance test and without complications. Living donor islet transplantation could cure one insulin dependent diabetes mellitus patients with a single donor. There are some advantages in the living donor islet transplantation: (a) living donor can alleviate the issue of donor shortage; (b) highly potent islets can be isolated from a living donor; and (c) the recipient can be treated with immunosuppressant and controlled blood glucose level tightly prior to the transplantation. These are important factors in overcoming the obstacles limiting islet transplantation. We believe that the living donor islet transplantation may become an additional option in treating insulin-dependent diabetes. PMID- 17130576 TI - DiaPep277 preserves endogenous insulin production by immunomodulation in type 1 diabetes. AB - DiaPep277 is an immunomodulatory peptide that arrests beta cell destruction in mouse models of type 1 diabetes mellitus (T1DM). This article extends an original pilot observation to two studies of 61 patients (age > 16 years), diagnosed with T1DM within 6 months, and with measurable beta cell function. Patients were treated with placebo (n = 27) or 1.0 mg DiaPep277 (n = 34). After 13 months, 1.0 mg Dia Pep277 treatment significantly (P = 0.02) preserved beta cell function as compared to the control with a trend for reduced HbA1c. This was achieved without an increase in insulin dose in the DiaPep277 group and with excellent safety. DiaPep277-treated patients also had fewer Th1 DiaPep277-specific T cells. PMID- 17130577 TI - Cord blood islet autoantibodies are related to stress in the mother during pregnancy. AB - A 2-month psychological questionnaire concerning pregnancy was answered by 20,920 nondiabetic mothers of singletons. Retrospective analysis showed increased levels of islet autoantibodies (IA) in 290 (1.4%) newborns. High IA levels in the child's cord blood correlated strongly with IA levels in the mother (GADA r = 0.91, P < 0.0001; IA-2A r = 0.75, P = 0.0001). High IA levels were found in newborns whose mothers during pregnancy had been more worried than usual (P = 0.04), had worried that the child would be sick (P = 0.01) or not survive (P = 0.002), or had experienced serious life events, like "serious accident in the family" (P < 0.0001) or "experienced violence" (P = 0.02). Associations with increased worries by the mother remained in newborns with high type 1 diabetes mellitus (T1DM)-human leukocyte antigen (HLA) risk, but not in non-HLA risk children. The prospective follow-up of these children will determine the importance of this early IA for postnatal islet autoimmunity, type 1 diabetes, or both. PMID- 17130578 TI - Is it dietary insulin? AB - In humans the primary trigger of insulin-specific immunity is a modified self antigen, that is, dietary bovine insulin, which breaks neonatal tolerance to self insulin. The immune response induced by bovine insulin spreads to react with human insulin. This primary immune response induced in the gut immune system is regulated by the mechanisms of oral tolerance. Genetic factors and environmental factors, such as the gut microflora, breast milk-derived factors, and enteral infections, control the development of oral tolerance. The age of host modifies the immune response to oral antigens because the permeability of the gut decreases with age and mucosal immune response, such as IgA response, develops with age. The factors that control the function of the gut immune system may either be protective from autoimmunity by supporting tolerance, or they may induce autoimmunity by abating tolerance to dietary insulin. There is accumulating evidence that the intestinal immune system is aberrant in children with type 1 diabetes (T1D). Intestinal immune activation and increased gut permeability are associated with T1D. These aberrancies may be responsible for the impaired control of tolerance to dietary insulin. Later in life, factors that activate insulin-specific immune cells derived from the gut may switch the response toward cytotoxic immunity. Viruses, which infect beta cells, may release autoantigens and potentiate their presentation by an infection-associated "danger signal." This kind of secondary immunization may cause functional changes in the dietary insulin primed immune cells, and lead to the infiltration of insulin reactive T cells to the pancreatic islets. PMID- 17130579 TI - Probiotics for the prevention of beta cell autoimmunity in children at genetic risk of type 1 diabetes--the PRODIA study. AB - The final aim of the PRODIA study is to determine whether the use of probiotics during the first 6 months of life decreases the appearance of type 1 diabetes mellitus (T1DM)-associated autoantibodies in children with genetic risk for T1DM. A pilot study including 200 subjects was planned to show whether the use of probiotics during the first 6 months of life is safe and feasible. The prevalence of autoantibodies among the study subjects at 6, 12, and 24 months of age was at levels close to the expected and the clinical follow-up did not either indicate problems in the feasibility of the study. PMID- 17130580 TI - Thiazolidinediones may not reduce diabetes incidence in type 1 diabetes. AB - Thiazolidinediones improve glycemic control by reducing insulin resistance. Some studies have demonstrated that troglitazone had a preventative effect on diabetes in NOD (non-obese diabetic) mice. One of the mechanisms proposed for the prevention of diabetes by thiazolidinediones is an effect on T-helper 1/T-helper 2 (Th1/Th2) balance. In this article, we attempted to clarify whether pioglitazone is also effective in preventing diabetes as compared to metformin, which has no immunological effect. Female NOD mice were administered pioglitazone or metformin orally, and the insulitis score, cytokines secreted from splenocytes, cytokine expression levels in the pancreas, and the incidence of diabetes after acceleration by cyclophosphamide were analyzed. We could not find any advantage of pioglitazone in preventing Th1 skewing and the development of diabetes over metformin. Therefore, further research should take place before the application of thiazolidinediones to human slowly progressive insulin-dependent diabetes mellitus (SPIDDM) patients. PMID- 17130581 TI - Mechanisms mediating anti-CD3 antibody efficacy: insights from a mathematical model of type 1 diabetes. AB - Anti-CD3 antibody therapy, a promising clinical approach for the treatment of type 1 diabetes (T1D), was investigated using a mathematical model of T1D in the female nonobese diabetic (NOD) mouse. Analyses of model simulation results indicate that, in addition to the known direct effects of anti-CD3 antibody on T lymphocytes, two additional mechanisms are required for sustained disease remission: (a) rapid regrowth of healthy beta cells following clearance of islet inflammation and (b) enhanced regulatory T cell activity and/or phenotypic changes in antigen presenting cells (APCs) that promote a stable regulatory environment in the pancreas. PMID- 17130582 TI - Why diabetes incidence increases--a unifying theory. AB - There is a wide spectrum within the diabetes syndrome. Type 1 diabetes may have a slow progression with good residual insulin secretion and without autoantibodies, while phenotypic type 2 diabetes may have autoantibodies. A single patient may have traits of both types of diabetes. Their incidence increases in parallel. The etiology is mainly unknown, but environmental factors play an important role in genetically predisposed individuals. The search for just one single cause of manifest diabetes may be confusing. Different mechanism may be important in different parts of the world. Furthermore, certain mechanisms may lead to islet inflammation while other/additional mechanisms may increase insulin demand and cause insulin deficiency with manifestation of clinical diabetes. Several hypothesis of etiology may fit different parts of the disease process. Thus, increased hygiene may contribute to an imbalance of the immune system, facilitating autoimmune reactions when virus infections, or proteins like cow's milk or gluten, provoke. Increased insulin demand because of rapid growth, or insulin resistance caused by stress, infections, puberty, etc., lead to beta cell stress, antigen presentation and may cause both an autoimmune reaction in genetically predisposed individuals, and insulin deficiency leading to manifest diabetes in individuals who have lost beta cell function. Vitamins may modulate the immune process, but we know too little to give vitamin substitution. However, we do know that low physical exercise, obesity, and stress, increases insulin demand resulting in insulin deficiency. Now we can therefore intervene to prevent the diabetic syndrome. PMID- 17130583 TI - Overcoming the challenges now limiting islet transplantation: a sequential, integrated approach. AB - Steady improvements in islet cell processing technology and immunosuppressive protocols have made pancreatic islet transplantation a clinical reality for the treatment of patients with Type 1 diabetes mellitus (T1DM). Recent trials are showing that improved glycemic metabolic control, prevention of severe hypoglycemia, and better quality of life can be reproducibly achieved after transplantation of allogeneic islets in patients with unstable T1DM. Despite these encouraging results, challenges ahead comprise obtaining adequate islet cells for transplant, enhancing islets engraftment, sustaining beta cell mass and function over time, and defining effective immune interventions, among others. In order to overcome the current hurdles to the widespread application of islet transplantation there is a need for implementation of integrated, sequential therapeutic approaches. PMID- 17130584 TI - Tendinitis: the Achilles heel of quinolones! AB - We present a case series of two patients who presented to the emergency department with spontaneous bilateral Achilles tendon rupture associated with the use of ciprofloxacin. Tendinitis and tendon rupture are now well recognised but rare complications of treatment with quinolone antimicrobials. The emergency department is an important setting for both surveillance and detection of adverse events associated with drug treatment. PMID- 17130585 TI - Brugada-like early repolarisation pattern associated with acute pericarditis. AB - Two cases of acute pericarditis presented with interesting electrocardiograms resembling Brugada-like or early repolarisation patterns. This report emphasises that proper interpretation of the electrocardiogram in patients with ST-segment elevation assists the clinician in arriving at the correct diagnosis in making appropriate diagnostic and therapeutic decisions, and also that the saddleback type ST-segment elevation cannot be a sensitive finding for the Brugada syndrome. PMID- 17130586 TI - Isolated Horner's syndrome caused by intraoral gunshot. AB - A 45-year-old man developed Horner's syndrome after sustaining an intraoral gunshot in a suicide attempt. Examination did not show any major vascular injury or other neurological symptoms. Radiograph of the skull and helical computed tomography angiography of the brain and neck detected a bullet embedded in the interior of the left internal carotid artery (ICA) sheath. Surgical examination showed an intact left ICA. Horner's syndrome resulting from intraoral trauma rarely presents as an isolated sign and is generally associated with carotid injuries. It may herald a life-threatening artery injury and needs urgent investigation. PMID- 17130587 TI - An eye for trouble: orbital cellulitis. AB - Patients with orbital cellulitis present to emergency departments occasionally. Symptoms usually develop rapidly, with patients being distressed by painful ocular movements and systemic upset. The case of a 24-year-old man who had a 1 month gradual history of intermittent periorbital swelling after a flu-like illness, and subsequently developed a large intracranial extradural abscess eroding through the temporal bone, ultimately requiring neurosurgical intervention is presented. Although orbital and periorbital swelling is common after acute sinusitis, cellulitis and intracranial abscess are rare but potentially life threatening and sight threatening. Accurate diagnosis is therefore of great importance. In this case, the patient presented with few clinical signs but relevant pathology. The importance of assessing ocular movement, which is a major clinical abnormality indicating orbital disorder and thus an aid to accurate diagnosis, should be highlighted. PMID- 17130588 TI - Blunt traumatic dissection of the proximal left anterior descending artery. AB - Coronary artery dissection secondary to blunt chest trauma is a rare occurrence recently reported in the Emergency Medicine Journal. The case here is reported of a left anterior descending artery dissection after blunt chest trauma to illustrate a different, more precipitous presentation of this condition after severe trauma. This case report highlights how clinical history can alert to the presence of this complication and how electrocardiography is fundamental to risk stratification after trauma, and discusses management alternatives for this condition in the setting of multi-trauma. PMID- 17130590 TI - A life in the day of a duty consultant. PMID- 17130591 TI - Consent for emergency care research: the Mental Capacity Act 2005. PMID- 17130592 TI - An evaluation of emergency medicine investigators' views on open access to medical literature. AB - BACKGROUND: Scientists and governmental agencies have called for free universal access to research publications via the internet--open access. OBJECTIVES: To examine the current medical literature reading practices of emergency medicine investigators (EMIs) and their views towards open access. METHODS: Surveys were mailed to the 212 corresponding authors of all original research articles published in years 2002 and 2003 in the Annals of Emergency Medicine, Academic Emergency Medicine and The Journal of Emergency Medicine. RESULTS: The most commonly read forms of medical literature reported by the 129 (61%) EMI respondents were hard-copy medical journals and online literature review services. 59% of EMIs were in favour of open access; 58% stated they would read a wider variety of medical literature; 21% believed open access would improve the quality of publications and 39% thought it would decrease the quality. When asked how a US 1500 dollars fee for open access would affect their ability to publish research, 69% said it would greatly impede and 19% said it would slightly impede their research. CONCLUSIONS: Despite concerns that open access may impede their ability to publish research and decrease the quality of publications, most EMIs surveyed favoured open access. They believed open access would increase and broaden their medical literature reading. PMID- 17130593 TI - Knowledge of and attitudes towards resuscitation in New Zealand high-school students. AB - BACKGROUND: Introducing cardiopulmonary resuscitation (CPR) training in the high school curriculum has been widely recommended as a long-term strategy to educate the wider community. Although CPR has been included in the New Zealand school curriculum, it is listed as an optional subject only. AIM: To assess the attitude towards and knowledge of CPR in 16-17-year-old high-school students in New Zealand. METHODS: Questionnaires were administered to 494 students aged 16-17 years across six high schools in Wellington, New Zealand. Both knowledge and attitude were evaluated in the questionnaire. RESULTS: Students showed poor theoretical knowledge, with a mean (SD) score of 5.61 (2.61) out of a maximum score of 18. Although there was no difference between male and female students, those who had received previous first-aid training (70%) showed greater knowledge (6.04 (2.56)) than their untrained counterparts (4.91 (2.24); p = 0.001). Those students with a positive attitude towards CPR and first-aid training (63%) acquired a higher knowledge score (6.12 (2.4)) than those with a negative attitude (17%; 4.65 (2.5); p = 0.001). Students with negative associations were also less likely to want to learn more about CPR and first aid (11%) when compared with those with positive associations (92%), and indicated less willingness to perform CPR on a stranger (negative v positive, 47% v 70%). CONCLUSIONS: These findings suggest that although most high-school students are willing and motivated to learn CPR, a smaller percentage of students had a negative attitude towards CPR that would act as a barrier to future learning or performance of resuscitation. Introducing CPR training to high schools is still recommended; however, this study shows the need to associate this training with positive references in an attempt to assist those for whom negative attitude may present as a barrier to learning and retaining CPR knowledge. PMID- 17130594 TI - Unilateral flail chest is seldom a lethal injury. AB - BACKGROUND: The chest cage is a common target for traumatic damage. Although relatively rare, it is considered to be a serious condition with significant reported mortalities. As most flail injuries are accompanied by severe extrathoracic injuries, it is often difficult to pinpoint a single injury responsible for the patient's death. AIM: To investigate the factors related to mortality when flail injury is diagnosed. METHODS: Data from the Israel National Trauma Registry between 1998 and 2003 included 11,966 chest injuries (262 flail chest injuries) out of a total of 118,211 trauma hospitalisations. Mortality figures were analysed to determine which factors, singly or in combination, influenced flail chest mortality. RESULTS: Road crashes accounted for most flail injuries (76%). The total mortality was 54 (20.6%) of 262 patients with flail chest injuries. 13 (20.4%) of the deaths occurred soon after admission to the emergency room and 37 (68.5%) within the first 24 h. Mortality in moderate to severe injuries (injury severity score (ISS) 9-24) was 3.6% and that in critical injuries 28.5% (ISS >24). Mortality increased with age: 17% in those aged <45 years, 22.1% in those between 45 and 64 years and 28.8% in those >65 years. Age remained a risk for inpatient death when adjusted for severity. Mortality in isolated unilateral flail injury was not more than 6%. Total mortality for traumatic brain injury (TBI) and flail was 34%. Flail, TBI and other major injuries increased the mortality to 61.1%. CONCLUSIONS: Advanced age is associated with higher mortality. Isolated unilateral bony cage instability infrequently leads to death in patients who make it to the emergency department but rather its combination with additional extrathoracic trauma. PMID- 17130595 TI - Validity of the Manchester Triage System in paediatric emergency care. AB - OBJECTIVE: To assess the validity of the Manchester Triage System (MTS) in paediatric emergency care, using information on vital signs, resource utilisation and hospitalisation. METHODS: Patients were eligible if they had attended the emergency department of a large inner-city hospital in The Netherlands from August 2003 to November 2004 and were <16 years of age. A representative sample of 1065 patients was drawn from 18,469 eligible patients. The originally assigned MTS urgency levels were compared with resource utilisation, hospitalisation and a predefined reference classification for true urgency, based on vital signs, resource utilisation and follow-up. Sensitivity, specificity and percentage of overtriage and undertriage of the MTS were calculated. RESULTS: The number of patients who used more than two resources increased with a higher level of MTS urgency. The percentage of hospital admissions increased with the increase in level of urgency, from 1% in the non-urgent patients to 54% in emergent patients. According to the reference classification, the sensitivity of the MTS to detect emergent/very urgent cases was 63%, and the specificity was 78%. Undertriage occurred in 15% of patients, of which 96% were by one urgency category lower than the reference classification. Overtriage occurred in 40%, mostly in lower MTS categories. In 36% of these cases, the MTS classified two or more urgency categories higher than the reference classification. CONCLUSIONS: The MTS has moderate sensitivity and specificity in paediatric emergency care. Specific modifications of the MTS should be considered in paediatric emergency care to reduce overtriage, while maintaining sensitivity in the highest urgency categories. PMID- 17130596 TI - Review of calls to NHS Direct related to attendance in the paediatric emergency department. AB - OBJECTIVE: To examine the outcomes of calls to NHS Direct (NHS-D) in relation to attendance at the accident and emergency (A&E) department. DESIGN: A prospective collection of data about consecutive calls to NHS-D North West Coast was matched with attendances at the A&E department over a period of 3 months. SETTING: NHS-D Regional Trust and a large urban paediatric A&E department. PATIENTS: Children and young adults aged <16 years living in local postal code areas. MAIN OUTCOME MEASURES: To examine (1) whether advice given by NHS-D was followed and (2) the differences in disease severity and necessity of attendance of patients referred by NHS-D and those referred by general practitioners and self-presenters. RESULTS: The relationship between the advice given and subsequent action is complex. Only 70% of calls advised to attend the A&E department did so. A further 1% (176) were advised not to attend the A&E department did in fact attend the department. Patients referred by NHS-D represented only 3.2% of department attendances. There was little difference in the triage categories of the presenting groups, but there were significantly less admissions (p<0.01) in the NHS-D group. CONCLUSIONS: Delivering telephone advice about illness severity in children is difficult as visual clues are so important. More collaborative prospective studies are needed, including with primary care, to understand families' choices, and to refine and assess NHS-D's ability to discriminate those requiring further clinical assessment. PMID- 17130597 TI - Ocular injuries in patients with major trauma. AB - AIM: To study the epidemiology of ocular injuries in patients with major trauma in the UK, determining the incidence and causes of ocular injuries, and their association with facial fractures. METHODS: A retrospective analysis of the Trauma Audit Research Network database from 1989 to 2004, looking at data from 39,073 patients with major trauma. RESULTS: Of the 39,073 patients with major trauma, 905 (2.3%) patients had associated ocular injuries and 4082 (10.4%) patients had a facial fracture (zygoma, orbit or maxilla). The risk of an eye injury for a patient with a facial fracture is 6.7 times as that for a patient with no facial fracture (95%, confidence interval 5.9 to 7.6). Of the patients with major trauma and an eye injury, 75.1% were men, and the median age was 31 years. 57.3% of ocular injuries were due to road traffic accidents (RTAs). CONCLUSION: The incidence of ocular injuries in patients with major trauma is low, but considerable association was found between eye injuries and facial fractures. Young adults have the highest incidence of ocular injury. RTAs are the leading cause of ocular injuries in patients with major trauma. It is vital that all patients with major trauma are examined specifically for an ocular injury. PMID- 17130598 TI - Use of the emergency department by Polish migrant workers. AB - Following the accession of Poland to the European Union in 2004, large numbers of Polish migrant workers have come to the UK. We describe how this migration has impacted on an emergency department in the West Midlands, and suggest advice that should be given to such visitors about how to use the NHS. PMID- 17130599 TI - Use of non-invasive ventilation in UK emergency departments. AB - AIM: To describe the current use of non-invasive ventilation in UK emergency departments. METHODS: A structured questionnaire was sent to all UK emergency departments assessing 25,000 new patients annually. RESULTS: 222 of 233 departments completed the questionnaire. 148 currently use non-invasive ventilation (NIV). Most used NIV for either cardiogenic pulmonary oedema (n = 128) or chronic obstructive pulmonary disease (n = 115). Only 49 departments have protocols for NIV use and 23 audited practice. CONCLUSION: NIV is commonly used in UK emergency departments. Practices vary significantly. One solution would be the development of guidelines on when and how to use NIV in emergency medicine practice. PMID- 17130600 TI - The safety of single-physician procedural sedation in the emergency department. AB - BACKGROUND: Previous research has shown the safety of procedural sedation in the emergency department in university settings involving multiple emergency physicians. OBJECTIVE: To examine sedation in the emergency department conducted by a single emergency physician with monitoring by the emergency nurse. METHODS: The Procedural Sedation in the Community Emergency Department Registry is a prospective observational database of procedural sedation cases directed by the emergency physicians. Among other parameters, the registry tracts whether emergency physicians or emergency nurses monitored patient sedation. The incidence of complications and outcomes were compared between these two monitoring groups. RESULTS: 1028 procedural sedations were performed on 977 patients at 14 sites. In 885 (86.1%) cases the emergency physician directed the sedation, and performed the procedure with monitoring by the emergency nurse. Complications occurred in 42 (4.1%) patients, 35 (4.0%) EN monitored patients and 6 (4.2%) EP monitored patients (p>0.7). Procedures were successful in 863 (97.5%) cases monitored by emergency nurses and in 140 (97.9%) patients monitored by emergency physicians (p>0.7). CONCLUSION: Procedural sedation in the emergency department performed by a single emergency physician is safe and effective. PMID- 17130601 TI - Faculty/College examinations: fitness for purpose. PMID- 17130602 TI - Diagnosis and management of common maxillofacial injuries in the emergency department. Part 2: mandibular fractures. AB - The majority of mandibular fractures occur in young males aged 16-30 years. As with all traumas, the history and examination cannot be over emphasised as diagnoses can be made with these tools. It is important when taking an accurate history to ascertain the mode and mechanism of injury (fall, punch, road traffic accident), and the magnitude and direction of the force involved (high or low velocity impact). PMID- 17130603 TI - Best evidence topic report. Silver sulphadiazine cream in burns. AB - A short-cut review was carried out to establish whether silver sulphadiazine cream is better than normal dressing in promoting healing without infection. A total of 410 papers were found using the reported searches, of which 12 showed the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. It is concluded that although there is evidence of antibacterial effect, there is no direct evidence of improved healing or reduced infection. PMID- 17130605 TI - Best evidence topic report. Procedural sedation for cardioversion. AB - A short-cut review was conducted to establish whether any of the available drugs used for procedural sedation in patients with tachydysrhythmias are safer or more effective than the alternatives. In all, 135 papers were found using the reported searches, of which 7 presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. It is concluded that propofol, methohexital, thiopentone and etomidate all seem to be good choices. PMID- 17130606 TI - Best evidence topic report. The relationship between post-traumatic stress disorder and affective disorder. AB - A short-cut review was carried out to establish whether the presence of affective disorder in victims of assault alters the chances of developing post-traumatic stress disorder (PTSD). In all, 133 papers were found using the reported searches, of which 8 presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. It is concluded that PTSD and affective disorder are related and often occur at the same time. PMID- 17130607 TI - Retrieval medicine: a review and guide for UK practitioners. Part 1: clinical guidelines and evidence base. AB - It has been proposed that formalisation of training to encompass prehospital and retrieval medicine should be considered in the UK, using those currently involved in immediate care as the core providers of these services.(1) Although there is an overlap in some aspects of "prehospital" and "retrieval" medicine, there are some distinct differences, both in terms of the skill base and service provision required. Retrieval medicine is the term used to indicate the use of an expert team to assess, stabilise and transport patients with severe injury or critical illness. Implicit in this process is the early provision of specialised advice to the health providers at the patient's side. In the UK, there is currently no national and often no regional strategy to coordinate the provision of secondary retrieval services for critically ill patients. International models do exist, which may be of help in this respect. PMID- 17130608 TI - Retrieval medicine: a review and guide for UK practitioners. Part 2: safety in patient retrieval systems. AB - Retrieval and transfer of critically ill and injured patients is a high risk activity. Risk can be minimised with robust safety and clinical governance systems in place. This article describes the various governance systems that can be employed to optimise safety and efficiency in retrieval services. These include operating procedure development, equipment management, communications procedures, crew resource management, significant event analysis, audit and training. PMID- 17130609 TI - Predictors of ambulance use in patients with acute myocardial infarction in Australia. AB - AIMS: To determine ambulance transport rates and investigate predictors for ambulance use by patients with acute myocardial infarction (AMI) in Australia. METHODS: A prospective, cross-sectional descriptive survey using structured interviews. It included patients who were admitted to two hospitals (Western, Bendigo, Melbourne, Victoria, Australia) with AMI between 1 October 2004 and 31 March 2005, and data were collected by semistructured interview and medical record review. Data were analysed by descriptive statistics, univariate and multivariate analysis using SPSS. RESULTS: 105 patients were interviewed. 48 (46%) participants called for an ambulance as their initial medical contact. Participants who called for an ambulance had a shorter interval between symptom onset and presentation to hospital than those who did not (non-ambulance participants)(median 2.1 v 7.8 h; p = 0.001). Predictors of ambulance transport were older age (p = 0.008), symptom onset on the weekend (p = 0.022), presence of sharp chest pain (p = 0.011), self-administered anginine (p = 0.007), symptom onset at home (p = 0.027) and having a lower income (37.5 degrees C and at least one constitutional symptom and one respiratory symptom, recruited during the survey period. Influenza and other respiratory viruses were identified using polymerase chain reaction (PCR) on throat swabs. Basic individual information was collected through a standard form. Of 173 ILI cases enrolled, 74 tested positive for one virus, and two tested positive for two viruses. Overall, 33.5% of the cases were positive for influenza viruses, 5.2% for adenoviruses, 3.5% for parainfluenza viruses, 1.7% for coronaviruses, and 1.2% for the respiratory syncitial virus. The proportion of influenza virus detection was higher in the 'high influenza activity' period. The distribution of viral agents varied across age groups, influenza viruses being more likely to be detected in younger patients. Viral pathogens were identified in less than 50% of ILI cases occurred during a high activity influenza season. The detection of other than influenza viruses was sporadic, without evidence of large outbreaks due to specific agents. PMID- 17130657 TI - Surveillance of influenza-like illness in England and Wales during 1966-2006. AB - We report surveillance data collected since 1966 from a general practice database in England and Wales. Incidence rates of influenza-like illness (ILI) peaked during the winter of 1969/70, and were then followed by a decade of heightened activity. There has since been a gradual downward trend of ILI, interspersed with winters of heightened activity; since 1999/2000, the incidence of ILI has been at its lowest for 40 years. We argue that the decade following the herald waves of the pandemic could be equally important for the planning of healthcare services in the community. PMID- 17130658 TI - Laboratory diagnosis of Lyme borreliosis at the Portuguese National Institute of Health (1990-2004). AB - Lyme borreliosis is considered to be an emerging infection in some regions of the world, including Portugal. The first Portuguese human case of Lyme borreliosis was identified in 1989. Since 1999, this disease is considered a notifiable disease (DDO) in Portugal, but only a few cases are reported each year, which does not allow consistent analysis of risk factors and the impact on public health. In this study the authors analyse the data available at the Centre for Vectors and Infectious Diseases Research (CEVDI) laboratory, at the Instituto Nacional de Saude Dr. Ricardo Jorge (National Institute of Health, INSA) during the past 15 years (1990-2004) and evaluate them against the registry of national reported cases (1999-2004). Serological tests were the basis for laboratory diagnosis. Data on year of diagnosis, sex, age, geographical origin and clinical signs are available for 628 well documented Portuguese positive cases. The number of cases per year varied between 2 and 78, with the highest number of cases reported in 1997. Of the positive cases, 53.5% were female and the age group most affected was 35-44 years old. Neuroborreliosis was the most common clinical manifestation (37.3%). Human cases were detected in 17 of the 20 regions of Portugal, and the highest number of laboratory confirmed cases were from the Lisbon district. The comparison of the number of notified cases and the number of positive cases confirmed by our laboratory show that Lyme borreliosis is clearly an underreported disease. Due to the scattered distribution of the positive cases and the low prevalence of the tick species Ixodes ricinus, the most effective prevention measure for Lyme borreliosis in Portugal is education of the risk groups on how to prevent tick bites. PMID- 17130659 TI - A measles outbreak in children under 15 months of age in La Rioja, Spain, 2005 2006. AB - This paper describes a measles outbreak in La Rioja, Spain, which began in December 2005 and mainly affected children under 15 months of age who were not yet immunised with MMR vaccine. The measles cases were detected by the mandatory reporting system, under which laboratories must report every confirmed measles case. Cases were classified in accordance with the National Measles Elimination Plan: suspected and laboratory-confirmed. In the period 14 December 2005 to 19 February 2006, 29 suspected cases of measles were investigated, and 18 were confirmed. The mean incubation period was 13.8 days (range: 9 to 18). Of the 18 confirmed cases, only two were in adults. MMR vaccination was recommended for all household contacts, as well as for children aged 6 to 14 months who attended the daycare centres where the cases had appeared. At these centres, the second dose of MMR was administered ahead of schedule for children under three years of age. It was recommended that the first dose of MMR vaccine be administered ahead of schedule for all children aged 9 to 14 months. During an outbreak of measles, children aged 6 months or older, who have not previously been vaccinated against measles, mumps and rubella, should receive a first dose as soon as possible, and those who have had a first dose should receive a second dose as soon as possible, provided that a minimum of one month has elapsed between the two doses. PMID- 17130660 TI - Measles elimination 2010 target: the need to meet the specific risk group. PMID- 17130661 TI - Atypical D-FISH patterns of BCR/ABL gene rearrangements in 169 chronic myeloid leukemia patients. AB - The Philadelphia (Ph) chromosome, a hallmark chromosomal anomaly observed in 95 percent of chronic myeloid leukemia (CML) cases, is known to involve the Abelson (ABL) proto-oncogene on chromosome 9 and the breakpoint cluster region (BCR) gene on chromosome 22, producing BCR/ABL mRNA encoding an abnormal tyrosine kinase protein. In the process of generating BCR-ABL fusion, the deletion of residual BCR or ABL occurs in 15-30 percent of CML patients. In addition, some rearrangements are complex, and do not yield the ABL/BCR fusion due to the involvement of a third chromosome in the rearrangement. The possible role of these deletions and complex rearrangements in disease outcome is an ongoing topic of research. We report our results of cytogenetic analysis with GTG banding and fluorescence in situ hybridization using dual color dual fusion probe (D-FISH) from Vysis Inc, USA in 169 (109 male and 60 female) CML patients registered at The Gujarat Cancer and Research Institute (GC and RI) from April 2004 to December 2005. GTG banding was carried out in 123 cases having analyzable metaphases. Of these 123 cases, D-FISH revealed atypical signal patterns in 57 patients (46%), and 12 cases revealed additional complex translocations indicative of disease progression. Out of 57 cases with atypical FISH patterns, 22 included metaphase FISH results, and the rest had only interphase FISH performed. In addition to the hallmark Philadelphia chromosome, other chromosomal aberrations in CML revealed heterogeneity of molecular events. Pooling of more data may lead to identification of new CML sub-groups and hence help in the analysis of clinical trials. Patients enrolled in our prospective study of prognostic significance will be followed up for disease free and overall survival in correlation with ABL BCR deletion status. PMID- 17130662 TI - CytoAccess, a relational laboratory information management system for a clinical cytogenetics laboratory. AB - We developed a CytoAccess laboratory management system based on the widely used Microsoft Access software to facilitate data processing, result reporting, and quality management for a full-service cytogenetics laboratory. The CytoAccess system consists of four functional modules. The data entry module is for logging in patient information. The result entry module is used to generate chromosome, fluorescent in situ hybridization (FISH), and array comparative genomic hybridization (aCGH) reports. The administrative module enables periodic monitoring of quality control and quality improvement (QA/QI) parameters and produces billing forms. The maintenance module allows users to update clinical demographics, report templates, code tables, and to refresh data links. We have integrated into this system over 15,000 chromosome and FISH results from prenatal, postnatal, and cancer cases for the past six years. This system is cost effective, user-friendly, flexible in updating, and potentially adaptable for data mining. PMID- 17130663 TI - Evidence for non-HFE linked hemochromatosis in Asian Indians. AB - BACKGROUND: Hereditary hemochromatosis is commonly due to two HFE1 (Histone Family E1) gene mutations - H63D and C282Y. Mutations in the Asian Indians are less well studied. AIMS: The aim of this preliminary study was to find out the prevalence of HFE gene mutations in nonviral liver cirrhosis patients. SETTINGS AND DESIGN: Unexplained liver cirrhosis cases with transferrin saturation> 45%, attending the gastroenterology clinic in the years 2004 and 2005 were subjects of the prospective study. Asymptomatic individuals with negative family history of hemolytic anemia or liver disease served as controls. MATERIALS AND METHODS: The clinical presentation was recorded in the patients. Transferrin saturation was estimated by standard colorimetric technique. The two common mutations in HFE1 gene and Y250X mutation of TFR (transferrin receptor) gene were studied by polymerase chain reaction based methods. RESULTS: A majority of the cases were sporadic, but family history was positive in four patients. In one family with multiple affected members, there was clear evidence of autosomal dominant inheritance. Seven out of 31 (22.6%) of unexplained cirrhosis cases were positive for mutations. One was homozygous for H63D. In healthy controls, prevalence was 8.1% (6/74). None of the patients or controls was positive for C282Y mutation of HFE1 or Y250X of TFR gene. CONCLUSIONS: Thus, in a number of cases of hemochromatosis in Indians, a gene with dominant inheritance may be involved in causation of the phenotype. The prevalence of HFE mutations in Indians is comparable to that reported from neighboring countries. It is worth studying other mutations in HFE gene and other iron overload genes in cryptogenic cirrhosis cases. PMID- 17130664 TI - Study of health problems and nutritional status of tea garden population of Assam. AB - BACKGROUND: Assam is the highest tea producer state in the country. There is scarcity of reliable information on health and nutritional status among tea garden population of Assam to enable initiating public health response to their health needs. AIMS: To describe health problems and nutritional status among tea garden population of Assam. SETTINGS AND DESIGN: Community-based cross-sectional survey in eight randomly selected tea gardens of Dibrugarh district of Assam. MATERIALS AND METHODS: Socio-demographic and behavioral characteristics of participants were recorded. Health problems and nutritional status were assessed through medical examination, evaluation of medical records, anthropometry and laboratory investigations. STATISTICAL ANALYSIS: Percentage prevalence; Chi square test was applied wherever applicable. RESULTS: Out of 4,016 participants, 1,863 were male and 2,153 were female. They were mostly illiterate and nearly 52.9% (1,197 of 2,264) of adults were manual workers in the garden. Alcohol and oral tobacco use were common. Prevalence of underweight among children was 59.9% (357 of 596) and thinness among adults was 69.9% (1,213 of 1,735). Anemia was widespread. Worm infection (65.4%, 217 of 332); skin problems; respiratory infections, including tuberculosis; filariasis were present in a significant way. Children suffered more in various diseases. Major noncommunicable diseases like hypertension, stroke were emerging in the community and were associated with modifiable risk factors like alcohol and tobacco use. CONCLUSION: Health status of the population can be ameliorated through better hygienic practices, environmental sanitation, creating health awareness, nutritional intervention and overall improvement of socioeconomic conditions of the population. PMID- 17130665 TI - Newborn care practices in an urban slum of Delhi. AB - BACKGROUND: Despite efforts by government and other agencies, neonatal morbidity and mortality continues to be high in India. Among other reasons, newborn care practices are major contributors for such high rates. AIMS: To find out the newborn care practices including delivery practices, immediate care given after birth and breast-feeding practices in an urban slum of Delhi. SETTINGS AND DESIGN: Community based, cross-sectional survey in a resettlement colony (a type of urban slum). MATERIALS AND METHODS: Semi-structured, pre-tested schedule was used to interview 82 mothers of newborns in the study area. STATISTICAL ANALYSIS: Data was analyzed using Epi - info version 6.04. Fischer exact test and chi2 test were applied. A P value of less than 0.05 was considered significant. RESULTS AND CONCLUSION: More than half i.e. 26 (56.1%) of home deliveries, which were mostly conducted by dais (24, 91.3%) or relatives in 4 (8.7%) of home deliveries. Bathing the baby immediately after birth was commonly practiced in 38 (82.6%) of home deliveries. Finger was used to clean the air passage in most of the home deliveries (29, 63%). About 61% (28) of home delivered newborns were not weighed at birth. Rooming in was practiced in majority of the cases. A few of home delivered neonates (12) were given injection tetanus toxoid by unqualified practitioners. Use of clip, band or sterile thread to tie the cord and no application to the cord was significantly higher in institutional deliveries. Breast milk as the first feed was significantly more in institutional deliveries. There is an urgent need to reorient health care providers and to educate mothers on clean delivery practices and early neonatal care. PMID- 17130666 TI - Resident psychiatrists as assessors for lectures in continued medical education in psychiatry. AB - OBJECTIVES: We examined the quality of instruction in a continued medical education course and the correspondence between the residents and lecturers evaluations of the program. MATERIALS AND METHODS: Resident psychiatrists and instructors completed structured evaluation forms immediately following each lecture in a psychiatry course for one academic year. RESULTS: Residents' and lecturers' evaluations of goal achievement, but not general ratings of lecture quality correlated positively. Instructors' enthusiasm, clarity and appropriateness of subject matter and encouragement of independent thinking, but not audio-visual aids significantly correlated with resident's positive evaluations. CONCLUSIONS: Ongoing evaluation of classroom lectures by resident psychiatrists may provide valuable feedback to instructors and impact the quality of medical education. The "classic" role of the teacher organization and enthusiasm are the most important components of quality education. PMID- 17130667 TI - An unusual case of systemic lupus erythematosus mimic: disseminated gastric signet ring cell carcinoma. PMID- 17130668 TI - Melatonin in pathogenesis and therapy of cancer. AB - Melatonin is a neuroendocrine hormone secreted by the pineal gland to transduce the body's circadian rhythms. An internal 24 hour time keeping system (biological clock) regulated by melatonin, controls the sleep-wake cycle. Melatonin production is a highly conserved evolutionary phenomenon. The indole hormone is synthesized in the pinealocytes derived from photoreceptors. Altered patterns and/or levels of melatonin secretion have been reported to coincide with sleep disorders, jetlag, depression, stress, reproductive activities, some forms of cancer and immunological disorders. Lately, the physiological and pathological role of melatonin has become a priority area of investigation, particularly in breast cancer, melanoma, colon cancer, lung cancer and leukemia. According to the 'melatonin hypothesis' of cancer, the exposure to light at night (LAN) and anthropogenic electric and magnetic fields (EMFs) is related to the increased incidence of breast cancer and childhood leukaemia via melatonin disruption. Melatonin's hypothermic, antioxidant and free radical scavenging properties, attribute it to an immunomodulator and an oncostatic agent as well. Many clinical studies have envisaged the potential therapeutic role of melatonin in various pathophysiological disorders, particularly cancer. A substantial reduction in risk of death and low adverse effects were reported from various randomized controlled trials of melatonin treatment in cancer patients. This review summarizes the physiological significance of melatonin and its potential role in cancer therapy. Furthermore, the article focuses on melatonin hypothesis to represent the cause-effect relationship of the three aspects: EMF, LAN and cancer. PMID- 17130669 TI - Reduction of disulfide bridges in the lumenal domain of ATF6 in response to glucose starvation. AB - Mammalian transcription factor ATF6 is constitutively synthesized as a type II transmembrane protein embedded in the endoplasmic reticulum (ER). Upon ER stress ATF6 is transported to the Golgi apparatus where it is cleaved to release its cytoplasmic domain. This is then translocated into the nucleus where it activates transcription of ER-localized molecular chaperones and folding enzymes to maintain the homeostasis of the ER. We recently found that, owing to the presence of intra- and intermolecular disulfide bridges, ATF6 occurs in unstressed ER in monomer, dimer and oligomer forms. Disulfide-bonded ATF6 is reduced on treatment of cells with various chemical ER stress inducers, and only the reduced monomer ATF6 reaches the Golgi apparatus. In this study, we evoked ER stress under more physiological conditions, namely, glucose starvation, and analyzed its consequence for ATF6 activation. Glucose starvation activated ATF6 and induced the ER chaperone BiP, albeit weakly. ATF6 was thus dissociated from BiP, transported to the Golgi apparatus, and cleaved. Glucose starvation enhanced the synthesis of ATF6 approximately two-fold, probably via transcriptional induction. Importantly, reduction of disulfide bridges and transport of reduced monomer occurred in response to glucose starvation. We conclude that ER stress-induced reduction of ATF6 represents a general feature of the ATF6 activation process. PMID- 17130670 TI - Lack of enhanced effect of antipsychotics combined with fluvoxamine on acetylcholine release in rat prefrontal cortex. AB - We have shown that coadministration of sulpiride and fluvoxamine preferentially increases the release of dopamine in the prefrontal cortex. To study the possible role of the cortical cholinergic system in this effect, we combined several other antipsychotic drugs with fluvoxamine and examined the effects on acetylcholine release in rat prefrontal cortex. Risperidone and clozapine significantly increased the release of acetylcholine but sulpiride did not, and fluvoxamine did not enhance the effects of the antipsychotics. These results further support the previous suggestion that the cortical dopamine system plays an important role in the effects of antipsychotic drugs administered in combination with fluvoxamine. PMID- 17130671 TI - Single-dose intravenous simvastatin treatment attenuates renal injury in an experimental model of ischemia-reperfusion in the rat. AB - The effect of acute pretreatment with a single dose of simvastatin (1 mg/kg, i.v.; 30 min before ischemia) on renal dysfunction caused by ischemia-reperfusion (I/R) injury in the rat was investigated. I/R injury was induced by clamping both renal vascular pedicles for 45 min, followed by 4 h of reperfusion with saline (2 ml/kg per hour). Simvastatin significantly improved both parameters of glomerular and tubular dysfunction (e.g., creatinine levels and fractional excretion of Na(+), respectively) and especially improved the histological score, compared to control I/R-injured rats treated with saline or 10% DMSO only. PMID- 17130672 TI - Comparison of four rate-correction algorithms for the ventricular repolarization period in assessing net effects of IKr blockers in dogs. AB - The utility of corrected and uncorrected QT interval changes for assessing net repolarization delay by I(Kr) (a rapid component of delayed rectifier K(+) currents) blockers was assessed in halothane-anesthetized dogs using the electrocardiogram and monophasic action potential (MAP) recordings with electrical ventricular pacing. Intravenous administration of dl-sotalol (0.2 - 2 mg/kg) prolonged the MAP duration and RR interval, while terfenadine (3 mg/kg) increased the MAP duration but transiently shortened RR interval. The order of correlation coefficient between the MAP duration at a pacing cycle length of 400 ms and MAP duration itself or that with arithmetical correction was uncorrected > Van de Water = Matsunaga > Fridericia > Bazett. These results suggest that Matsunaga's and Van de Water's formulae would better predict the net repolarization delay in the in vivo canine model. Also, the risk of drug candidates that may prolong the QT interval should be judged by change in uncorrected QT interval as well as corrected QT interval. PMID- 17130673 TI - Changes in emotional behavior of mice in the hole-board test after olfactory bulbectomy. AB - The most consistent behavioral change caused by olfactory bulbectomy (OBX) is a hyperemotional response to novel environmental stimuli. The aim of this study was to characterize the emotional behavior of OBX mice using the hole-board test. After the olfactory bulbs were lesioned, sham and OBX mice were housed in single cages for 14 days. The number of head-dips in the hole-board test in single housed OBX mice was significantly greater than that in single-housed sham mice. The head-dipping behaviors in single-housed sham and OBX mice were reversed by treatment with diazepam, a typical benzodiazepine anxiolytic. (+/-)-8-Hydroxy-2 (di-n-propylamino) tetraline hydrobromide (8-OH-DPAT), a selective 5-HT(1A) receptor agonist that has a non-benzodiazepine anxiolytic-like effect, and (+)-4 [(aR)-a-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N-diethyl benzamide (SNC80), a delta-opioid-receptor agonist, also significantly reversed the number of head-dips in single-housed sham and OBX mice. In conclusion, we suggest that the single-housed OBX mice showed heightened emotional behavior (e.g., increase in head-dipping behavior) in the hole-board test. In addition, we suggest that the hyperemotional behavior characterized by head-dipping behavior in OBX mice was selectively reversed by benzodiazepine and non-benzodiazepine anxiolytics. PMID- 17130674 TI - Contribution of extracellular signal-regulated kinase to UTP-induced interleukin 6 biosynthesis in HaCaT keratinocytes. AB - UTP causes interleukin (IL)-6 production via mRNA expression through P2Y(2)/P2Y(4) receptors in human HaCaT keratinocytes. In the present study, we analyzed the mechanism of UTP-induced IL-6 production in these cells. UTP, an agonist of P2Y(2)/P2Y(4) receptors, induced phosphorylation of extracellular signal-regulated kinase (ERK) in a concentration- and time-dependent manner. PD98059, a MEK (mitogen-activated protein kinase kinase) inhibitor, and BAPTA-AM [O,O'-bis(2-aminophenyl)ethyleneglycol-N,N,N',N'-tetraacetic acid, tetraacetoxymethyl ester], an intracellular Ca(2+) chelator, reduced UTP-induced ERK phosphorylation and IL-6 mRNA expression. 2-APB [(2 aminoethoxy)diphenylborane], an inositol 1,4,5-trisphosphate (IP(3))-receptor antagonist, inhibited UTP-induced IL-6 mRNA expression; and the action of A23187, a Ca(2+) ionophore, resembled the action of UTP. In contrast, protein kinase C (PKC) downregulation and pertussis toxin did not affect UTP-induced IL-6 mRNA expression, suggesting that PKC and G(i) are not involved in the UTP-induced IL-6 production. However, AG1478, an epidermal growth factor (EGF)-receptor inhibitor, partially decreased UTP-induced ERK phosphorylation and IL-6 expression. These results suggest that UTP-induced IL-6 production is in part mediated via phosphorylation of ERK through G(q/11)/IP(3)/[Ca(2+)](i) and transactivation of the EGF receptor. PMID- 17130675 TI - Short-term and long-term FK506 treatment alters the vascular reactivity of renal and mesenteric vascular beds. AB - The aims of this study were to investigate the role of endothelin-1 in FK506 induced hypertension and vascular dysfunction of rats treated with the drug for 8 (short-term) or 30 (long-term) days and to measure malondialdehyde levels in the kidneys. Kidney and mesentery of rats were perfused. In the short-term treated groups, there was no significant change in systolic blood pressure. The response to noradrenaline only in renal vascular beds was significantly increased by FK506 and this increase was prevented by Bosentan. FK506 had no significant effect on sodium nitroprusside-induced vasodilation in comparison with solvent in both vascular beds. Bosentan failed to prevent these responses. In the long-term treated groups, at the end of the treatment with FK506, there was a significant increase in blood pressure, but no change in the response to noradrenaline in either kidneys or mesentery. The increase in blood pressure was prevented by bosentan treatment. FK506 increased malondialdehyde levels in the kidneys of the rats from only the long-term treated groups. Bosentan did not change this increase. Our results indicated that endothelin-1 plays a key role in the FK506 induced change in vascular reactivity to noradrenaline in renal vascular beds and drug-induced hypertension in the rats. There was no relationship between oxidative stress and FK506-induced hypertension. PMID- 17130676 TI - Osteopontin: a potential biomarker for successful bee venom immunotherapy and a potential molecule for inhibiting IgE-mediated allergic responses. AB - Venom immunotherapy (VIT) is proven to be curative for insect allergy, but the mechanisms and the biomarkers associated with clinical efficacy remain elusive. We report herein the discovery of a leading candidate biomarker, osteopontin (OPN), for VIT. From cDNA microarray and clustering analyses, an increased expression of OPN was found in patients who completed 5-6 years of VIT and discontinued therapy for 3-6 years as compared with the untreated group. A significantly higher level of serum OPN was found in the completed treatment group as compared with the untreated group. Following VIT, kinetically increased levels of OPN associated with reduced venom specific IgE levels were noted in subjects with large local allergic reactions to venom. These findings together with the fact that OPN is involved in Th1-associated immune response strongly suggest a role of OPN as a functional biomarker for VIT. PMID- 17130677 TI - Microarray-based identification of novel biomarkers in asthma. AB - Bronchial asthma is a complicated and diverse disorder affected by genetic and environmental factors. It is widely accepted that it is a Th2-type inflammation originating in lung and caused by inhalation of ubiquitous allergens. The complicated and diverse pathogenesis of this disease yet to be clarified. Functional genomics is the analysis of whole gene expression profiling under given condition, and microarray technology is now the most powerful tool for functional genomics. Several attempts to clarify the pathogenesis of bronchial asthma have been carried out using microarray technology, providing us some novel biomarkers for diagnosis, therapeutic targets or understanding pathogenic mechanisms of bronchial asthma. In this article, we review the outcomes of these analyses by the microarray approach as applied to this disease by focusing on the identification of novel biomarkers. PMID- 17130678 TI - Respiratory viral infections and early asthma in childhood. AB - Respiratory viral infections profoundly influence the disease activity of wheezing illnesses and asthma in early childhood. Viral bronchiolitis shares many features with asthma and a subset of children develop recurrent wheezing after their initial illness. Recently mechanisms for virus-induced exacerbations of childhood asthma are beginning to be focused on and defined. Viruses cause systemic immune activation and also produce local inflammation. These factors are likely to affect airway pathogenesis leading to airway narrowing, an increase in mucus production, and eventually bronchospasm, and airway obstruction. These new insights related to the pathogenesis and disease activity are likely to provide new targets for the therapy and prevention of early asthma in childhood. PMID- 17130679 TI - Effect of antihistamine eye drops on the conjunctival provocation test with Japanese cedar pollen allergen. AB - BACKGROUND: Approximately 16.2% of the Japanese population suffer from cedar pollinosis, with various manifestations such as ophthalmic, laryngo-pharyngeal and skin symptoms in addition to nasal symptoms. Thus, the annual pollen season is an agonizing period for patients. No study has reported symptoms and their clinical courses after conjunctival provocation with purified cedar pollen allergen Cry j1 as well as suppression of these allergen-induced ocular symptoms by antihistamine eye drops. METHODS: Nine patients with Japanese cedar pollinosis who had no nasal or ocular symptoms were included in the present study, after obtaining informed consent in writing. 1) Purified cedar pollen allergen Cry j1 was instilled in the left eye and phosphate-buffered saline (PBS) in the right eye as a control. 2) Levocabastine hydrochloride ophthalmic suspension and ketotifen fumarate ophthalmic solution were respectively instilled in the left and right eyes, which were then challenged with the allergen. Ocular symptoms after provocation with the allergen were recorded through the clinical course. RESULTS: Pollen allergen-induced ocular symptoms were itching and hyperemia of the palpebral conjunctiva, and itching lasted for more than 5 hours. Moreover, preadministration of antihistamine eye drops suppressed the increases in the ocular symptom scores, eliminating itching within 1 hour. Allergen provoked not only ocular symptoms but also nasal symptoms in 77.8% of patients. CONCLUSIONS: Preadministration of antihistamine eye drops suppressed the symptoms induced by the allergen, which suggests that this is an effective early therapy for Japanese cedar pollinosis, if it is started before the pollen season. However, self protection by patients using a mask may not be effective enough to suppress nasal symptoms during the pollen season, requiring them to additionally wear glasses to avoid exposure to the allergen. PMID- 17130680 TI - Omalizumab is effective and safe in the treatment of Japanese cedar pollen induced seasonal allergic rhinitis. AB - BACKGROUND: Seasonal allergic rhinitis (SAR) induced by Japanese cedar pollen is a substantial problem in Japan. Omalizumab, a novel humanized monoclonal anti immunoglobulin E (IgE) antibody, has already been proven to reduce symptoms associated with SAR. We investigated the safety and efficacy of omalizumab in the treatment of patients with Japanese cedar pollen-induced SAR compared to placebo. METHODS: A randomized, placebo-controlled, double-blind study was conducted in 100 Japanese patients with a history of moderate-to-severe SAR induced by Japanese cedar pollens. Omalizumab (150, 225, 300, or 375mg) or placebo was administered subcutaneously every 2 or 4 weeks based on serum total IgE and body weight at baseline. The primary efficacy variable was the mean of daily nasal symptom medication scores (sum of the daily nasal symptom severity score and daily nasal rescue medication score) during the treatment period. Secondary efficacy variables included the daily ocular symptom medication score and related variables. RESULTS: Primary and all secondary efficacy variable scores were significantly lower in the omalizumab group than in the placebo group (P < .01). Serum free IgE levels markedly decreased in the omalizumab group and were associated with clinical efficacy. The overall incidence of injection site reactions was higher in the omalizumab group than in the placebo group; however, the adverse reaction profile was similar between the two groups when excluding injection site reactions. No anti-omalizumab antibodies were detected. CONCLUSIONS: Omalizumab was effective and safe in the treatment of SAR induced by Japanese cedar pollen. PMID- 17130681 TI - Effects of salmeterol xinafoate and fluticasone propionate on immunological activation of human cultured mast cells. AB - BACKGROUND: The clinical efficacy of combination therapy comprising a long acting beta(2)-agonist (LABA) and corticosteroid is widely recognized for the treatment of adult asthma. Here we examine the effect of salmeterol xinafoate (SX) and fluticasone propionate (FP) alone and in combination on the immunological activation of human cultured mast cells (HCMC)in vitro. METHODS: HCMC were passively sensitized with IgE antibody and then activated by challenging with anti-IgE antibody. The effect of drugs on the activation of mast cells was examined by measuring the amount of released chemical mediators (histamine, leukotrienes (LT) and prostaglandin D(2) (PGD(2))) and granulocyte macrophage colony stimulating factor (GM-CSF). RESULTS: The release of each chemical mediator was inhibited by 10-9-10-8M SX but not by 10-10-10-7M FP. The production of GM-CSF was inhibited by a concentration of 10-8M in both drugs and the inhibition was augmented by combined treatment with 10-11M of each drug. CONCLUSIONS: The immunological release of chemical mediators (histamine, LT, PGD(2)) from HCMC was inhibited by SX but not by FP. SX and FP inhibited the production of GM-CSF by HCMC and both drug showed synergistic inhibition in the production of GM-CSF. PMID- 17130682 TI - A prospective survey on safety of sustained-release theophylline in treatment of asthma and COPD. AB - BACKGROUND: Theophylline is a useful drug for the treatment of asthma. The Asthma Prevention and Management Guidelines (JGL) recommend use of sustained-release theophylline products as controllers and of injectable aminophylline products as relievers. The Global Initiative for Asthma: Global Strategy for Asthma Management and Prevention, the NHLB/WHP Workshop Report 1995 (GINA, 1995) and guidelines in Western countries recommend sustained-release theophylline, but not as positively as in the JGL. The aim of this survey was to determine the occurrence of serious adverse reactions. METHODS: The survey was conducted in 66 institutions staffed by physicians certified by the Japanese Society of Allergology (JSA). The target diseases were asthma and COPD including chronic bronchitis and pulmonary emphysema, which are indications for use of sustained release theophylline products in Japan. RESULTS: 3,921 patients were included in the safety evaluation. No serious adverse reactions were observed among the patients in this survey, although 54 patients (1.38%) exhibited non-serious adverse reactions. The incidence of adverse reactions was found to be high in patients who had begun use of sustained-release theophylline products at the time of registration in this survey, and in patients who were concomitantly taking macrolide antibiotics. CONCLUSIONS: The present survey demonstrates that sustained-release theophylline is safe, as long as used appropriately, although adverse reactions tend to develop early after initiation of administration. PMID- 17130683 TI - Effects of KP-496, a novel dual antagonist for leukotriene D4 and thromboxane A2 receptors, on contractions induced by various agonists in the guinea pig trachea. AB - BACKGROUND: A dry powder inhaler of KP-496 is currently in clinical development in Japan as an anti-asthmatic agent. The aim of this study was to evaluate the in vitro pharmacological profile of KP-496. METHODS: The antagonistic activities of KP-496 for leukotriene (LT) D(4) and thromboxane (TX) A(2) receptors were examined using the LTD(4)- and U46619-induced contractions of the isolated guinea pig trachea. The selectivity of KP-496 was examined using various agonist-induced contractions in the isolated guinea pig trachea. RESULTS: KP-496 produced parallel rightward shifts of the LTD(4) and U46619 concentration-response curves in a concentration-dependent manner. Schild plot analyses of the antagonistic activities of KP-496 demonstrated that it is a competitive antagonist for LTD(4) and TXA(2) receptors with pA(2) values of 8.64 and 8.23, respectively. The LTD(4) antagonistic activity of KP-496 was comparable to that of pranlukast and zafirlukast but was more potent than that of montelukast. The TXA(2) antagonistic activity of KP-496 was comparable to that of seratrodast. KP-496 and seratrodast also inhibited the prostaglandin (PG) D(2)- and PGF(2alpha)-induced contractions of the isolated guinea pig trachea. KP-496 had no effect on the histamine-, acetylcholine-, serotonin- and substance P-induced contractions of the isolated guinea pig trachea. CONCLUSIONS: These results indicate that KP-496 is a selective dual antagonist for LTD(4) and TXA(2) receptors. LTD(4) and TXA(2) play important roles in asthma, and antagonists for these mediators are being used for the treatment of asthma. Thus, KP-496 is expected to become a novel potent therapeutic agent for asthma. PMID- 17130684 TI - Urticarial reaction caused by ethanol. AB - BACKGROUND: We report a case of an urticarial reaction after drinking alcohol beverages. The patient was a 47-year-old man suffering urticarial and anaphylactoid reaction to alcohol for two years. These reactions were observed at every alcohol beverages intake. CASE SUMMARY: We performed a prick test with diluted ethanol, alcohol beverages and their metabolites (acetaldehyde, acetic acid). Only acetic acid showed a positive result. Oral challenge test with diluted-ethanol caused pruritus and swelling of his lips. An oral challenge test with 8% diluted Shochu (Japanese distilled alcohol from rice or wheat) caused wheals on his upper back. DISCUSSION: Only acetic acid, a metabolite of alcohol, induced a positive prick test in the patient with alcohol-induced urticaria. This result was not observed in normal volunteers. An oral challenge test with diluted alcohol or Shochu showed a positive wheal reaction in a dose dependent-manner which suggests that urticaria seen in this patient might be induced by alcohol intolerance. However possible allergic reaction to acetaldehyde could not be excluded. PMID- 17130685 TI - Physical activity and breast cancer: a systematic review. AB - BACKGROUND: Many epidemiologic studies have found an association between physical activity and breast cancer risk, although this has not been a consistent finding. METHODS: Studies were identified through a systematic review of literature available on PubMed through February 2006. We included all cohort and case control studies that assessed total or leisure time activities in relation to occurrence or mortality of breast cancer. The fully adjusted risk estimates and 95% confidence intervals for the highest versus lowest level of activity were documented for each study as well as evidence for a dose-response relationship. Methodologic quality was also assessed. Due to statistical and methodologic heterogeneity among studies, we did not carry out statistical pooling. To draw conclusions, we performed a best-evidence synthesis taking study quality into account. RESULTS: Nineteen cohort studies and 29 case-control studies were evaluated. There was strong evidence for an inverse association between physical activity and postmenopausal breast cancer with risk reductions ranging from 20% to 80%. For premenopausal breast cancer, however, the evidence was much weaker. For pre- and postmenopausal breast cancer combined, physical activity was associated with a modest (15-20%) decreased risk. Evidence for a dose-response relationship was observed in approximately half of the higher-quality studies that reported a decreased risk. A trend analysis indicated a 6% (95% confidence interval = 3% to 8%) decrease in breast cancer risk for each additional hour of physical activity per week assuming that the level of activity would be sustained. CONCLUSIONS: There is evidence for an inverse association between physical activity and breast cancer risk. The evidence is stronger for postmenopausal breast cancer than for premenopausal breast cancer. PMID- 17130686 TI - Impact of the specificity of the exposure metric on exposure-response relationships. AB - BACKGROUND: Exposure misclassification may occur when nonspecific exposure indicators are used. Developing estimates of more specific measures may be difficult due to sampling limitations or a paucity of historical measurements and, thus, often requires substantial effort. We examine the impact on exposure response relationships of moving from 2 measures of exposure mixtures (dust, chlorophenols) to more specific exposure indicators (wood dust, pentachlorophenol, tetrachlorophenol) in a retrospective cohort. METHODS: The study population consisted of 26,847 male sawmill workers (> or =1 year employment between 1950 and 1995) with linkage to national cancer registries. A subcohort (n = 11,273 employed more than 1 day between 1985 and 1995) was linked to hospital discharge records. We evaluated the shape (log-linear vs log-log models), goodness of fit, precision, and expected versus observed attenuation of the exposure-response relationships. RESULTS: The correlation between the cumulative exposure indices was moderately high (dust/wood dust, r = 0.68; total chlorophenol/pentachlorophenol, r = 0.88; total chlorophenol/tetrachlorophenol, r = 0.78). An increase in chronic obstructive pulmonary disease hospitalizations was found with wood dust but not with total dust. Stronger associations for non Hodgkin lymphoma and kidney cancer incidence were observed with pentachlorophenol than with total chlorophenol; no association was observed with tetrachlorophenol. We observed greater attenuation than expected using total dust, but less than expected using total chlorophenol. CONCLUSIONS: The relationships between health outcomes were substantially attenuated when nonspecific exposure indicators were used. This study demonstrates the importance of developing exposure metrics as specific to the disease-causing agent as possible, particularly when the composition of mixed exposures varies by work areas. PMID- 17130687 TI - Neighborhood socioeconomic deprivation and residential instability: effects on incidence of ischemic heart disease and survival after myocardial infarction. AB - BACKGROUND: Previous literature has shown that neighborhood socioeconomic position influences the risk of ischemic heart disease, but little is known about the mechanisms linking the residential context to ischemic heart disease incidence and mortality. We examined whether neighborhood socioeconomic position and neighborhood residential stability (as a determinant of social interaction patterns) have an influence on ischemic heart disease risk. Moreover, we investigated whether dissimilar contextual influences operate at different stages of the disease process, ie, on incidence, 1-day case-fatality, and long-term survival after acute myocardial infarction (MI). METHODS: Using a large 27-year longitudinal cohort (baseline: 1 January 1996) defined in the Scania region, Sweden, we estimated multilevel survival models adjusted for individual sociodemographic factors and previous diseases of the persons. RESULTS: After adjustment, multilevel survival models indicated that the incidence of ischemic heart disease increased with neighborhood socioeconomic deprivation but was only weakly associated with neighborhood residential instability (for high vs low residential instability, hazard ratio = 1.2; 95% credible interval = 1.0-1.4). Conversely, beyond effects of individual and contextual socioeconomic circumstances and distance to the hospital, we saw a markedly higher 1-day case fatality (4.9; 1.8-15) and shorter survival time after MI among individuals still alive 28 days after MI (4.3; 1.2-17) in neighborhoods with a high versus low residential instability. CONCLUSIONS: Effects of residential instability on post MI survival may be mediated by the lower availability of social support in residentially unstable neighborhoods, suggesting a new class of intermediate processes that should be taken into account when investigating contextual influences on ischemic heart disease. Moreover, dissimilar contextual effects may operate at various stages of the disease process (ie, on incidence, case fatality, and survival after MI). PMID- 17130688 TI - Cumulative lead exposure and cognitive performance among elderly men. AB - BACKGROUND: Recent evidence suggests that cumulative lead exposure among adults in nonoccupational settings can adversely affect cognitive function. Which cognitive domains are affected has not been explored in detail. METHODS: We used nonlinear spline regressions and linear repeated-measures analysis to assess the association between scores on a battery of cognitive tests over time and both blood and bone lead concentrations in the Normative Aging Study, a cohort of community-dwelling elderly men. Bone lead was measured from 1991 through 1999 with K-shell x-ray fluorescence. A total of 1089 men with a mean (+/-standard deviation) age of 68.7 (+/-7.4) years with blood lead measurements, 761 of whom also had valid bone lead measurements, completed at least one of a battery of cognitive tests. Approximately 3.5 years later, 69% of the men had at least one repeat test. Cognitive testing was performed from 1993 through 2001. RESULTS: On a cross-sectional basis, there was little association between blood or bone lead and cognitive test scores. Change in performance over time on virtually all tests worsened as bone lead increased, with the most robust effects on performance and reaction time scores on visuospatial/visuomotor tests. CONCLUSIONS: Low-level cumulative exposure to lead in nonoccupational settings may adversely affect cognitive function, particularly in the visuospatial/visuomotor domain. PMID- 17130689 TI - Plasma selenium over time and cognitive decline in the elderly. AB - BACKGROUND: Because brain oxidative stress is a cause of cognitive impairment, selenium, which is an antioxidant, may protect against cognitive decline. The aim of the study was to examine whether declining selenium levels over time are associated with cognitive decline in a cohort of community-dwelling French elderly. METHODS: During 1991-1993, 1389 subjects (age 60-71 years) were recruited into a 9-year longitudinal study with 6 waves of follow-up. Cognitive functions were evaluated by neuropsychologic tests. To take into account the entire set of cognitive measurements and the within-subject correlations between measures, we analyzed mixed linear and logistic models to study associations between selenium change and cognitive decline. RESULTS: After controlling for potential confounders, cognitive decline was associated with decreases of plasma selenium over time. Among subjects who had a decrease in their plasma selenium levels, the greater the decrease in plasma selenium, the higher the probability of cognitive decline. Among subjects who had an increase in their plasma selenium levels, cognitive decline was greater in subjects with the smallest selenium increase. There was no association between short-term (2-year) selenium change and cognitive changes. CONCLUSION: Selenium status decreases with age and may contribute to declines in neuropsychologic functions among aging people. PMID- 17130693 TI - A new scoring system to predict fast-track failure in cardiac surgery. PMID- 17130694 TI - Inhaled nitric oxide: should it be used in acute respiratory distress syndrome? PMID- 17130695 TI - Fresh water versus salt water: when will the seas meet! PMID- 17130696 TI - Hypertonic saline for the treatment of intracranial hypertension: worth its salt. PMID- 17130697 TI - Tracheostomy: it's time to move from art to science. PMID- 17130698 TI - Family presence at resuscitation: what if? PMID- 17130699 TI - The fat ones fare well--but is it fair to compare? PMID- 17130700 TI - Intrapulmonary percussive ventilation and noninvasive positive pressure ventilation in patients with chronic obstructive pulmonary disease: "Strength through unity"? PMID- 17130701 TI - Donation after cardiac death in the United States: how to move forward. PMID- 17130702 TI - Severe acute exacerbation of chronic obstructive pulmonary disease: when are multiple-drug-resistant bacteria the culprits? PMID- 17130703 TI - Moral distress and the intensive care unit. PMID- 17130704 TI - Implementing strict glucose control: it is not that simple... PMID- 17130705 TI - Off-pump cardiac surgery and acute kidney injury. PMID- 17130706 TI - A new diagnostic test for an old diagnostic challenge: thyrotoxic periodic paralysis. PMID- 17130707 TI - Ventilatory management of acute lung injury by neurally adjusted ventilatory assist and positive end-expiratory pressure. PMID- 17130708 TI - A shocking surprise: a novel defibrillation protocol to treat cardiac arrest. PMID- 17130709 TI - Hypertonic saline: a marker for discrimination between a disrupted and intact blood-brain barrier? PMID- 17130711 TI - Protective effects of adenosine A2A agonist during hemorrhagic shock: a simple intervention may result in a complex response. PMID- 17130712 TI - High positive end-expiratory pressure, low tidal volume ventilatory strategy in persistent acute respiratory distress syndrome. PMID- 17130715 TI - Death by total parenteral nutrition: part deaux. PMID- 17130716 TI - Identifying factors that could account for differences in duration of mechanical ventilation between intermittent lorazepam- and propofol-treated patients. PMID- 17130717 TI - Oxime use after deliberate release. PMID- 17130719 TI - Role of cytokines in the pathogenesis of transfusion-related pulmonary edema. PMID- 17130721 TI - Emergency medicine resident interest in critical care fellowship training increases if provided United States certification pathway. PMID- 17130723 TI - Toward an accurate assessment of the adverse effects of packed red blood cell transfusions in cardiac surgery. PMID- 17130726 TI - Transverse colon mass. PMID- 17130727 TI - Investigating the whole gut. PMID- 17130728 TI - Do bones crack under the effects of inflammatory bowel disease in children? PMID- 17130729 TI - Inflammatory bowel disease in infants: the other "end of the beginning"? PMID- 17130730 TI - Evaluation of oral rehydration solution by whole-gut perfusion in rats: effect of osmolarity, sodium concentration and resistant starch. AB - BACKGROUND: Reduced osmolarity oral rehydration solution (ORS) improved small bowel absorption of fluid and electrolytes in segmental perfusion in experimental animals; this was borne out in clinical practice. Adding amylase-resistant starch (RS) to ORS is expected to increase colonic fluid absorption. This study used combined small and large bowel perfusion to evaluate combinations of reduced osmolarity and starch in ORS. METHODS: Single-pass steady-state perfusions of the whole gut at 30 mL/h, using the nonabsorbable marker C-polyethylene glycol 4000, were performed in Wistar rats after exposure to cholera toxin or Escherichia coli heat-stable enterotoxin (STa). RESULTS: Steady state was established within 90 minutes after commencing perfusion. Net secretion of water, sodium and chloride induced by cholera toxin was partially reversed by standard glucose-ORS (G-ORS). Substituting glucose in G-ORS with RS (RS-ORS) substantially increased net water absorption (P < 0.001) as did reduced osmolarity ORS (RO-ORS) (P < 0.001); addition of RS to RO-ORS further increased water absorption (P < 0.001). In STa treated intestine, RO-ORS and RS-ORS significantly improved water absorption compared to G-ORS (P < 0.005). RO- and RS-RO-ORS did not significantly augment net electrolyte absorption compared with G-ORS. RS-ORS was associated with highest net absorption of sodium and chloride compared with all other groups. CONCLUSIONS: RS increased net water (and sodium) absorption from isosmolar and reduced osmolar ORS consistent with increased absorption by the colon. RS in reduced osmolar ORS may have advantages to reduce severity of diarrhea and prevent hyponatremia in severe diarrhea and may be applicable to diarrhea of different etiologies. PMID- 17130731 TI - Does small intestinal atresia affect epithelial protein expression in human newborns? AB - OBJECTIVES: Bowel segments distal to a congenital intestinal obstruction have been suggested to be immature. In other words, luminal components such as amniotic fluid (before birth) and/or enteral nutrition (after birth) may be required to activate intestinal epithelial protein expression, thereby influencing epithelial differentiation. We investigated cell-type-specific protein expression proximal and distal to jejunal and ileal atresias in human newborns. PATIENTS AND METHODS: We immunohistochemically studied intestinal tissue specimens of 16 newborns who had undergone surgery for jejunal or ileal atresia. Sections were taken from both the proximal and distal sides of the atresias. RESULTS: For all patients, the enterocyte-specific markers lactase, sucrase-isomaltase, sodium glucose cotransporter 1, glucose transporters 2 and 5, intestinal fatty acid-binding protein and alkaline phosphatase were expressed at a mean 3 +/- 1 days after birth, both proximal and distal to jejunal and ileal atresias. Expression of goblet cell-specific markers mucin 2 and trefoil factor 3 and that of the Paneth cell marker lysozyme was maintained at either side of the atretic segment. CONCLUSIONS: With respect to the markers used, the human small intestinal epithelium is already differentiated shortly after birth. The absence of intestinal continuity in case of a jejunal or ileal atresia does not affect epithelial protein expression. This would seem to indicate that the developing small intestinal epithelium matures independently of luminal components. PMID- 17130732 TI - Human faecal microbiota develops the ability to degrade type 3 resistant starch during weaning. AB - BACKGROUND: Colonisation of the human colon starts immediately after birth. Bacterial composition is substantially influenced by the type of feeding. During weaning, microbiota diversifies considerably to finally approach the composition of that of adults. The aim of this study was to investigate the ability of colonic microbiota obtained from different age groups to ferment resistant starch (RS). METHODS: Faecal samples of breast-fed and formula-fed infants, infants at weaning, adults and elderly subjects were used as inocula. Fermentation experiments were performed by applying a standardised in vitro batch method. Fermentability was established by measuring both metabolite production and substrate degradation. An RS type 3 (RS-3) was used as substrate; its behaviour was compared with that of lactulose (positive control), whereas inoculum without substrate was used as negative control. RESULTS: Overall fermentation patterns clearly showed that the human microbiota of all age groups is able to degrade lactulose. In contrast, RS-3 was found resistant to the attack by microbiota of both breast-fed and formula-fed infants. Bacteria collected from infants at weaning were able to degrade RS-3 completely, but slower compared with adults. With increasing age, RS-3 fermentation was observed to be slightly retarded again. CONCLUSIONS: Human faecal microbiota of all age groups is able to ferment lactulose in vitro quickly and completely. The ability to degrade RS-3, however, is only established during weaning. Whether fermentation-related production of short-chain fatty acids from RS-3 and concurrent modifications of the microbiota can result in potential health benefits to the host at this stage of life needs to be elucidated. PMID- 17130733 TI - Efficacy of granulocyte apheresis in pediatric patients with ulcerative colitis: a pilot study. AB - OBJECTIVES: Granulocyte apheresis (GCAP), involving the removal of granulocytes from the blood, may improve clinical symptoms and facilitate a reduction in the dose of steroids in adult patients with ulcerative colitis. As a preliminary trial, GCAP was used to taper the dose of steroids in 4 pediatric patients with ulcerative colitis. METHODS: Three males and 1 female ranging from 11 to 17 years old were treated with GCAP once per week for 5 consecutive weeks/course. The ages of patients at clinical onset ranged from 8 to 12 years and the length of time from the clinical onset to GCAP treatment ranged from 28 to 58 months (median, 38.5 months). RESULTS: In 2 patients, symptoms and signs indicating disease activity improved after 2 courses of GCAP. Laboratory data and endoscopic findings also improved after treatment and the clinical efficacy was judged to be excellent in these patients. In 1 patient, GCAP improved laboratory and endoscopic hallmarks, but bloody stools persisted. Finally, the treatment was ineffective in the fourth patient who eventually underwent surgery. CONCLUSIONS: GCAP is effective in improving clinical symptoms and may play an important role in converting steroid therapy to other treatments in children with steroid refractory or steroid-dependent ulcerative colitis. PMID- 17130734 TI - The prevalence of long bone fractures in pediatric inflammatory bowel disease. AB - INTRODUCTION: The association of inflammatory bowel disease (IBD) with decreased bone mineral density is well recognized. In the adult population, up to 50% of IBD patients are reported to have osteopenia, correlating with an increase in the incidence of fractures as compared with controls. The aim of this study was to determine the prevalence of fractures in a pediatric population with IBD as compared with healthy sibling controls (SC). PATIENTS AND METHODS: The families of 209 patients with IBD were sent a questionnaire asking them to compare their children with IBD to a healthy sibling (non-IBD). RESULTS: Surveys were returned by 132 of the 209 families (63%). The sample characteristics of this sample closely resembled the overall clinic population for age (mean 14.3 vs 14.7 years), gender (53% vs 59% male) and diagnosis (58.1 vs 57.8 Crohn disease). Completed surveys described 263 children. Of the 132 with IBD 73 (55%) had Crohn disease, 52 (39%) had ulcerative colitis and 7 (6%) had indeterminate colitis. There were 76/132 males (age range, 4-18 years) with IBD and 64/131 males (age range, 1-26 years) in the sibling controls. Mean ages of the IBD sample 14.3 +/ .3 was compared with 13.9 +/- in SC. Of the total group, 73/263 (28%) reported ever having a fracture, 44 (60%) were siblings (SC), and 29 (40%) had IBD. Of the 29 children with IBD, 17 (59%) reported having a fracture after diagnosis including 2 who had fractures both before and after diagnosis. The total number of fractures reported was 96 (55 SC:41 IBD). CONCLUSION: In this survey, we found no statistically significant difference in the prevalence of fracture in IBD patients compared with their normal siblings. PMID- 17130735 TI - Characteristics of inflammatory bowel disease with onset during the first year of life. AB - BACKGROUND: Inflammatory bowel disease (IBD) is recognized in young children, however, only rare data on onset and evolution are available in children younger than 1 year. In the present clinical study, we aimed to analyze characteristics and clinical course of children with very early onset IBD. We were particularly interested in the relationship between bacterial infections and the use of antibiotics before the onset of IBD. PATIENTS AND METHODS: The IBD database of Necker-Enfants-Malades-Hospital was screened for patients with IBD with disease onset during the first year of life and a follow-up of at least 2.5 years. Ten patients were identified during the period 1996-2002. RESULTS: All patients presented with rectal bleeding and had colonic involvement. Four patients had definitive diagnosis of Crohn disease; ulcerative or indeterminate colitis was seen in 2 and 4 children, respectively. Five of the patients had a positive history of neonatal or early-onset bacterial infection with use of antibiotics before onset of IBD, 4 patients were still breastfed and 3 just weaned when GI symptoms started. Seven patients had a severe onset of disease requiring bowel rest, parenteral nutrition and steroid medication, followed by azathioprine or cyclosporine medication. Surgery was necessary in 3 of 10 patients. Disease relapses were frequent and observed in 8 of 10 children. DISCUSSION: Very early onset IBD may reflect a subgroup of patients characterized by a particular sensitivity to modifications of the intestinal flora. Neonatal IBD was most often severe in presentation and evolution. PMID- 17130736 TI - Enterobius vermicularis and colitis in Children. AB - OBJECTIVES: We observed a cohort of children presenting with rectal bleeding that were identified as having Enterobius vermicularis at colonoscopy and questioned the reliability of conventional diagnostic methods of identifying E. vermicularis. PATIENTS AND METHODS: The study was retrospective in nature and subjects were investigated by colonoscopy between May 1997 and December 1999. Patients were identified as having E. vermicularis infestation by direct visualisation of the adult worms at colonoscopy. Patients were treated with mebendazole and a record of their clinical response documented. RESULTS: A total of 180 colonoscopic examinations were performed during the study period. E. vermicularis was identified macroscopically in 31 cases (17.2%). The symptom profile of patients with E. vermicularis were abdominal pain, 19 of 26 (73%); rectal bleeding, 16 of 26 (62%); chronic diarrhoea, 13 of 26 (50%) and weight loss, 11 of 26 (42%). Ova cysts and parasites were identified in none of the saline swabs analysed in 20 patients. Sellotape testing was performed in only 4 patients and was negative in all. Of the 26 children, 21 (81%) demonstrated histopathological features of nonspecific colitis. There was clinical resolution of symptoms in 19 of 23 patients. CONCLUSIONS: We suggest that in patients with symptoms suggestive of inflammatory bowel disease, E. vermicularis infestation must be excluded as a common cause of nonspecific colitis. We also suggest that diagnostic tests such as saline swabs and Sellotape testing may be lacking in sensitivity. PMID- 17130737 TI - A new indirect chemiluminescent immunoassay to measure anti-tissue transglutaminase antibodies. AB - OBJECTIVES: Anti-tissue transglutaminase antibody (anti-tTG) determination using second-generation (human antigen) enzyme-linked immunoassays (ELISAs) is a very accurate test to diagnose celiac disease (CD). In this study, we compared 2 second-generation ELISAs (Celikey tTG; Pharmacia Diagnostics GmbH & Co, Freiburg, Germany, and QuantaLite; Inova Diagnostics, San Diego, CA) and antiendomysial antibodies (EMAs) with a new indirect chemiluminescence immunoassay (LIAISON tTG; DiaSorin S.p.A., Saluggia, Italy) in diagnosing and monitoring CD in children. PATIENTS AND METHODS: Antiendomysial antibodies, anti-tTGs and total immunoglobulin A were measured in the sera of 103 control children, 101 children with histologically proven CD and 31 CD children on gluten-free diet (GFD). RESULTS: Anti-tissue transglutaminase antibody mean levels were significantly higher in CD with respect to control or GFD children. The sensitivity value of EMAs, LIAISON tTG, Celikey tTG and QuantaLite in diagnosing CD was 97.7%, 97.0%, 94.1% and 98.0%, respectively, and the corresponding specificity values were 91.1%, 98.1%, 97.1% and 96.1%, respectively. The degree of mucosal destruction (Marsh criteria) was correlated with EMA semiquantification (P < 0.01) and with the circulating levels of anti-tTGs measured using LIAISON (P < 0.05) or QuantaLite (P < 0.01). Twenty-six CD children were followed up from 5 to 25 months after GFD. The circulating levels of anti-tTGs measured with any of the 3 assays significantly dropped after GFD. CONCLUSIONS: Anti-tissue transglutaminase antibody determination with second-generation ELISAs is as effective as EMAs for CD diagnosis. The novel chemiluminescent method described in the present paper for the detection of anti-tTGs in the diagnosis of CD had the highest sensitivity and specificity values. The anti-tTG test correlates with the degree of mucosal destruction and is suitable for verifying patient compliance to dietary treatment. PMID- 17130738 TI - Epithelial electrical resistance as a measure of permeability changes in pediatric duodenal biopsies. AB - OBJECTIVES: Intestinal permeability measured with medium-sized oral probes is increased in cystic fibrosis (CF) and celiac disease (CD), probably reflecting reduced tight junction resistance. The aim of this study was to evaluate whether square-pulse analysis of duodenal biopsies from children can be used to determine electrical tight junction resistance. METHODS: Intestinal biopsies from children with different stages of CD and from patients with CF were studied in a modified Ussing chamber. The epithelium was assumed to act as an electrical circuit consisting of a current generator parallel with a resistance and a capacitance. Subepithelial and epithelial resistances were determined by square-pulse analysis, and the generated current was calculated. RESULTS: Confirming data using permeability probes, reduced epithelial electrical resistance was found both in patients with CF and CD. Only the CF patients had reduced resting current as well. The secretagogues prostaglandin E2, cyclic adenosine monophosphate and acetylcholine increased the current in both control biopsies and biopsies with villous atrophy but had no significant effect on epithelial resistance. CONCLUSIONS: Measurement of electrical resistance in duodenal biopsies can be used as an alternative method of quantifying permeability in pediatric biopsies. PMID- 17130739 TI - Percutaneous endoscopic gastrostomy in children: a safe technique with major symptom relief and high parental satisfaction. AB - BACKGROUND: Percutaneous endoscopic gastrostomy (PEG) is widely used for establishing enteral feeding. The aim of this study was to assess immediate and long-term results after PEG insertion. PATIENTS AND METHODS: A total of 121 children were retrospectively reviewed. Median age was 2.4 years (range, 4 months 13.2 years) at the time of PEG insertion. Patient morbidity, indications for PEG, preoperative findings and perioperative complications were registered retrospectively. Parents/caregivers of 85 children were interviewed for long-term results. RESULTS: Perioperative complications were seen in 12%. Twenty-four percent died at a median of 15 months (range, 1.5 months-8 years) after PEG. Eighty-five families were interviewed with a median follow-up time of 5.6 years (range, 1-10 years). A substantial majority (94%) of parents/caregivers reported that the PEG had a positive influence on their child's situation, and 98% would have chosen PEG insertion again. Vomiting/retching improved in 61% of the children, and oral intake enhanced in 43%. Stoma-related complications were frequent (73%). The gastrostomy tube was permanently removed at a median of 3 years (range, 7 months-7.3 years) after PEG placement in 25%. Delayed closure of the gastrocutaneous fistula after gastrostomy removal occurred in 48% of them. Time from insertion to removal was not predictive of delayed closure. CONCLUSION: PEG is a safe technique for establishing enteral feeding, even in very sick children. Major complications are rare, although most children experience minor stoma-related problems. Parents/caregivers report that the gastrostomy is of great help for themselves and their child. PMID- 17130740 TI - Prospective analysis of nonadherence in autoimmune hepatitis: a common problem. AB - OBJECTIVES: To prospectively assess nonadherence to medications, the relationship between nonadherence and medical outcome and the relationship between a psychiatric risk factor (posttraumatic stress) and nonadherence in patients with a diagnosis of autoimmune hepatitis. PATIENTS AND METHODS: Data were obtained in children with autoimmune hepatitis, who had consented to prospective monitoring of adherence, during 1 year of follow-up in our pediatric liver program. An electronic monitoring device as well as posttransplant trough blood levels of tacrolimus was used to evaluate adherence. A validated self-report questionnaire was used to assess posttraumatic stress. The medical outcome measure was the maximal alanine transaminase (ALT) for each monitored patient. RESULTS: Of 37 pediatric patients, 34 (15 posttransplant) enrolled. Fourteen (41%) used the monitoring device as directed. Monitor readings ranged between 28% and 94% of optimal adherence (100%). No patient took the medications exactly as prescribed. Electronic monitoring device readings correlated inversely with maximal ALT (P = 0.03, r = -0.59), and were also correlated with the tacrolimus level variability as a measure of adherence (P = 0.04, r = -0.72). Posttraumatic stress disorder questionnaire scores were correlated with both measures of adherence (for electronic monitoring, P = 0.02, r = -0.70, for tacrolimus levels, P = 0.03, r = 0.62). CONCLUSIONS: Nonadherence to immunosuppressants was common in this cohort, and it correlated with higher maximal ALT. Nonadherence is therefore an important risk factor for poor outcome in patients with autoimmune hepatitis. Posttraumatic stress symptoms, which were correlated with nonadherence, may serve as a focus for adherence-improving interventions. PMID- 17130741 TI - Follow-up of children with autoimmune hepatitis treated with cyclosporine. AB - OBJECTIVES: To evaluate the effectiveness of cyclosporine in inducing and maintaining remission of the inflammatory process in autoimmune hepatitis, when used in combination with low doses of prednisone and azathioprine and to identify the prognostic factors associated with sustained remission. METHODS: Eighty-four patients with autoimmune hepatitis were consecutively recruited from 5 centers between January 1994 and March 2001. Cyclosporine was administered during the first 6 months. Thereafter, in patients with aminotransferase levels of lower than twice the normal values, prednisone and azathioprine were initiated. RESULTS: Normal aminotransferase levels were observed in 94.05% (79/84) of the patients, 72% of them within the first 6 months of treatment. Total serum bilirubin level of greater than 1.2 mg/dL and portal hypertension at diagnosis jointly predicted a significant delay in remission. Adverse effects related to cyclosporine remained mild and transient. Low doses of prednisone and standard doses of azathioprine were not implicated in relapse of the disease during the follow-up of any patient. CONCLUSIONS: This protocol allowed control of the liver inflammatory process and was well tolerated. The response to this immunosuppressive therapy can be predicted with accuracy. Factors delaying remission can be identified early at diagnosis and may contribute to the development of more effective treatment policies for this condition. PMID- 17130742 TI - Prognostic value of portal pressure at the time of Kasai operation in patients with biliary atresia. AB - AIM: To examine the prognostic value of portal pressure at the time of Kasai operation in patients with biliary atresia (BA). PATIENTS AND METHODS: All 127 patients with BA, born between 1991 and 1996, who underwent a Kasai operation in Bicetre Hospital were studied. Intraoperatively, the umbilical vein was catheterised and a portal pressure index (PPI), defined by the height of the physiologic saline column above the liver surface level, was measured. Overall patient survival, survival with native liver (SNL) and, in patients living with their native liver, survival without significant portal hypertension (SwsPHT) were calculated by Kaplan-Meier method and compared by univariate and multivariate analyses. RESULTS: Measurement of PPI was successful in 113/127 patients (89%) without complication. Median PPI was 15 cm (range, 5-35). Portal pressure index was positively correlated with age at surgery and histological severity of liver fibrosis. Ten-year overall patient survival was 78.2%; 10-year SNL was 35.7%. In patients living with their native liver, 5- and 10-year SwsPHT were 65.3% and 43.6%, respectively. Portal pressure index >15 cm, complete extrahepatic BA and polysplenia were independently associated with a worse SNL. Portal pressure index >15 cm and the absence of postoperative normalisation of bilirubin were independently associated with a worse SwsPHT. CONCLUSION: Biliary atresia patients with elevated portal pressure at the time of Kasai operation have lower chances of success of this procedure and a higher risk of developing portal hypertension, even if bilirubin levels normalise after the operation. Measurement of PPI was simple and safe and better predicted the postoperative outcome than did the histological scores of liver fibrosis. PMID- 17130743 TI - Decreased coenzyme Q10 concentration in plasma of children with cystic fibrosis. AB - OBJECTIVES: Coenzyme Q10 (CoQ10) is an effective lipophilic antioxidant and protects against lipid peroxidation by scavenging radicals. Patients with cystic fibrosis generally have fat malabsorption; thus, we hypothesized that overall plasma CoQ10 concentration in pediatric patients with cystic fibrosis might be diminished. Because these patients have increased oxidative stress due to chronic pulmonary inflammation, we also assumed that the oxidized form of CoQ10 might be relatively increased. PATIENTS AND METHODS: The total plasma CoQ10 levels and the oxidized and reduced form were measured by high-performance liquid chromatography in 30 children with cystic fibrosis (mean FEV1 % predicted = 88.5% +/- 18.7%) and 30 age-matched controls. RESULTS: Total plasma CoQ10 levels were significantly lower in the cystic fibrosis group as compared with the control group (0.87 +/- 0.42 micromol/L and 1.35 +/- 0.39 micromol/L, respectively; P < 0.001). When correcting for the lower serum cholesterol level in patients with cystic fibrosis, this difference remained significant: the CoQ10/cholesterol ratio (micromol/mol) was 268.8 +/- 136.7 and 334.0 +/- 102.9 in patients and controls, respectively (P < 0.05). However, the CoQ10 redox status was identical in patients and controls (86.4% +/- 7.1% and 85.4% +/- 7.3%, respectively). CONCLUSIONS: We found that the overall plasma CoQ10 concentration is lower in patients with cystic fibrosis, probably because of fat malabsorption. The CoQ10 redox status was not disturbed, indicating that CoQ10 could still be adequately regenerated in this group of patients with cystic fibrosis with mild-to-moderate pulmonary disease. PMID- 17130744 TI - Meal patterns, dietary fat intake and pancreatic enzyme use in preadolescent children with cystic fibrosis. AB - OBJECTIVES: To describe the usual pattern of intake and adherence to cystic fibrosis (CF) food and pancreatic enzyme replacement therapy (PERT) recommendations in preadolescent children with CF and pancreatic insufficiency (PI). METHODS: Children, 8 to 11 years old, with CF and PI were assessed for PERT adherence ratio (number taken/number prescribed per day) and energy and fat intake for 7 days (weighed food records), adherence to PERT and dietary recommendations, pulmonary function, growth status and stool fat malabsorption. Using the PERT adherence ratio, children were divided into good (> or =80%), moderate (60% to 79%) and poor (<60%) adherence groups. RESULTS: Of the 75 children (age, 9.3 +/- 1.0 years; forced expiratory volume in 1 s, 95 +/- 14% predicted), 61% consumed less than 120% of energy intake, and 72% consumed less than 40% of calories as fat recommended for CF. Using the PERT adherence ratio (mean +/- SD, 75 +/- 14%), 29%, 61% and 9% of children had good, moderate and poor adherence, respectively. Better adherence to PERT was significantly associated with missing fewer snacks, higher energy intake, greater fat content of snacks and poorer growth status. About 85% of the children were in the 500 to 4000 lipase unit (LU) of PERT (LU/g fat per day) and 500 to 2500 LU/kg per body weight per meal guidelines, whereas only 58% to 68% were within the guidelines for snacks. Eleven percent of children always took PERT at an inappropriate time after meals. CONCLUSIONS: Most preadolescent children with CF and PI did not meet dietary recommendations. Future educational and behavioral interventions focused on increasing PERT adherence with snacks, fat content of meals and snacks, appropriate timing of enzymes and adjustment of PERT and meal fat content are suggested. PMID- 17130745 TI - Dietary and circulating polyunsaturated fatty acids in cystic fibrosis: are they related to clinical outcomes? AB - OBJECTIVE: To assess the relationship between dietary intakes, plasma phospholipid (PL) fatty acid profile and clinical parameters in children with cystic fibrosis (CF) in comparison to healthy controls. PATIENTS AND METHODS: A cross-sectional survey including 37 patients with CF (ages 8.0 +/- 2.9 yrs) and a reference group of 68 healthy children (ages 8.0 +/- 0.7 yrs) was carried out by means of a food-frequency questionnaire. At enrollment, all subjects underwent blood sampling for plasma PL fatty acids (FA). In CF patients, pulmonary function tests (forced expiratory volume in 1 second and forced vital capacity), anthropometric measurements and the Shwachman score were also determined. RESULTS: In CF patients, mean z score for weight and height (-0.35 +/- 1.16 and 0.28 +/- 0.99) were lower than controls (0.83 +/- 1.73 and 0.55 +/- 1.11, respectively). Patients with CF showed higher energy intakes (110 +/- 43 kcal/d) compared with controls (75 +/- 22 kcal/d; P < 0.0001), with higher intake of total (saturated and monounsaturated) fats and lower intake of polyunsaturated FA (3.9 +/- 1.0% of total macronutrient intake vs 4.3 +/- 1.2%, P = 0.05). In CF patients, plasma and PL levels of linoleic and docosahexaenoic acids were lower, whereas those of arachidonic acid were similar compared with controls. The Shwachman score showed significant positive associations with plasma PL levels of arachidonic acid and total n-6 long-chain FA (r = 0.32, P = 0.05, and r = 0.35, P = 0.03, respectively). CONCLUSIONS: The data give suggestions that fat intake and CF-associated biomechanisms are bound in a vicious circle, concurring to create the clinical and biochemical picture of CF. The quantity and quality of fat supplementation in CF need careful attention to balance the fat supply with polyunsaturated FA. PMID- 17130746 TI - Acetate correction for postabsorption metabolism does not improve the [13C]mixed triacylglycerol breath test. AB - OBJECTIVE: The [C]mixed triacylglycerol (MTG) breath test is a noninvasive measure of fat digestion. After absorption and oxidation, C appears in breath CO2. Recovery is no more than 50% in healthy subjects because of sequestration of acetate in intermediary metabolism. The aims of this study were to investigate interindividual variation and postabsorptive metabolism of MTG using oral [1 C]acetate and to examine the use of correction factors to account for the "missing" label. PATIENTS AND METHODS: [C]mixed triacylglycerol and [1-C]acetate breath tests were performed on 8 healthy adults, 9 healthy children and 3 children with cystic fibrosis. Breath was sampled for 6 hours on each occasion. The enrichment of CO2 in breath was measured by isotope ratio mass spectrometry. Cumulative percentage dose recovered CO2 during the MTG test was corrected for label absorbed, but not completely oxidized using the cumulative percentage dose recovered during the acetate test. RESULTS: Mean recovery of C during the [C]MTG test with an acetate correction was close to 100% in healthy subjects: 103.1% (SD, 11.6%) in adults and 98.9% (SD, 30.3%) in children, but the wide variance indicated that some of the assumptions governing the use of acetate corrections with oral tracers may not be valid. CONCLUSION: The need to perform 2 tests, variation in physical activity between tests and differences in intermediary metabolism preclude the use of acetate correction factors when using [C]MTG to assess intraluminal fat digestion, especially in children. PMID- 17130747 TI - Effects of bovine alpha-lactalbumin and casein glycomacropeptide-enriched infant formulae on faecal microbiota in healthy term infants. AB - OBJECTIVE: Certain milk factors may promote the growth of a host-friendly gastrointestinal microbiota, for example, one that is predominated by bifidobacteria, a perceived health-promoting genus. This may explain why breast fed infants experience fewer intestinal infections than their formula-fed counterparts who are believed to have a more diverse microbiota, which is similar to that of adults. The effects of formulas supplemented with 2 such ingredients from bovine milk, alpha-lactalbumin (alpha-lac) and casein glycomacropeptide (GMP), on gut flora were investigated in this study. PATIENTS AND METHODS: Six week-old (4-8 wk), healthy term infants were randomised to a standard infant formula or 1 of 2 test formulae enriched in alpha-lac with higher or lower GMP until 6 months. Faecal bacteriology was determined by the culture-independent procedure fluorescence in situ hybridisation. RESULTS: There was a large fluctuation of bacterial counts within groups with no statistically significant differences between groups. Although all groups showed a predominance of bifidobacteria, breast-fed infants had a small temporary increase in counts. Other bacterial levels varied in formula-fed groups, which overall showed an adult-like faecal microflora. CONCLUSIONS: It can be speculated that a prebiotic effect for alpha-lac and GMP is achieved only with low starting populations of beneficial microbiota (eg, infants not initially breast-fed. PMID- 17130749 TI - Varicella-associated colitis in a 6-month-old infant. PMID- 17130748 TI - Erythropoiesis and myocardial energy requirements contribute to the hypermetabolism of childhood sickle cell anemia. AB - OBJECTIVES: We hypothesized that an elevated hemoglobin synthesis rate (SynHb) and myocardial oxygen consumption (MVO2) contribute to the excess protein and energy metabolism reported in children with sickle cell anemia. PATIENTS AND METHODS: Twelve children (6-12 years old) with asymptomatic sickle cell and 9 healthy children matched for age and sex were studied. Measurements were whole body protein turnover by [1-C]leucine, SynHb by [N]glycine, resting energy expenditure by indirect calorimetry and the systolic blood pressure-heart rate product used as an index of MVO2. Protein energy cost was calculated from protein turnover. Statistical analysis included Spearman correlations and partial correlation analyses. RESULTS: Although body mass index was significantly lower for sickle cell versus controls (P < 0.02), children with asymptomatic sickle cell had 52% higher protein turnover (P < 0.0005). Proportional reticulocyte count, SynHb, MVO2 and resting energy expenditure were also significantly higher in children with sickle cell (P < 0.01). Protein turnover correlated significantly with both SynHb (r = 0.63, P < 0.01) and reticulocyte percentage (r = 0.83, P < 0.0001). Partial correlation of these 3 variables showed reticulocyte percentage as the only variable to be significantly associated with protein turnover, even after adjusting for sickle cell anemia (P = 0.03). Partial correlation of log resting energy expenditure on MVO2 was significant, controlling for protein energy cost, sex and age (P = 0.03). CONCLUSION: These results indicate that metabolic demands of increased erythropoiesis and cardiac energy consumption account for much of the excess protein and energy metabolism in children with sickle cell anemia. PMID- 17130750 TI - Eosinophilic gastroenteritis presenting as acute gastric perforation. PMID- 17130751 TI - Spontaneous perforation of the bile duct in a toddler: the role of endoscopic retrograde cholangiopancreatography in diagnosis and therapy. PMID- 17130752 TI - A case of an upside-down stomach. PMID- 17130753 TI - Acquisition of competence in paediatric ileocolonoscopy with virtual endoscopy training. AB - This study prospectively compared the rates of skill acquisition in ileocolonoscopy in 2 consecutive groups of trainees in paediatric gastroenterology, with 1 cohort exposed to virtual endoscopy. All paediatric gastroenterology trainees rotating through our department during a 7-year period between 1997 and 2004 were formally assessed while performing ileocolonoscopies using a trainer case-by-case method. Fourteen consecutive trainees with no previous experience of ileocolonoscopy were assessed. Comparison of rates of skill acquisition and lesion recognition using multiple linear regressions revealed a significant acceleration of achievement of endoscopic goals (P < 0.0001) in the group with prior exposure to virtual endoscopy. PMID- 17130756 TI - Handle with Care: the American Nurses Association's campaign to address work related musculoskeletal disorders. AB - In response to the significant number and severity of work-related back injuries and other musculoskeletal disorders among nurses, the American Nurses Association (ANA) has launched the Handle With Care(R) campaign. The campaign seeks to build a health care industry-wide effort to prevent back and other musculoskeletal injuries. This is being done through developing partnerships and coalitions, education and training, increasing use of assistive equipment and patient handling devices, reshaping nursing education to incorporate safe patient handling, and pursuing federal and state ergonomics policy by highlighting technology-oriented safe-patient handling benefits for patients and nurses. In the absence of ergonomics regulations at national or state levels that protect health care workers, ANA has taken on alternative approaches to encourage a movement to control ergonomic hazards in the health care workplace and prevent back injuries among the nation's nursing workforce. PMID- 17130757 TI - Update on Handle with Care: the American Nurses Association's campaign to address work-related musculoskeletal disorders. PMID- 17130758 TI - Evidence-based practices for safe patient handling and movement. AB - Efforts to reduce injuries associated with patient handling are often based on tradition and personal experience rather than scientific evidence. The purpose of this article is to summarize current evidence for interventions designed to reduce caregiver injuries, a significant problem for decades. Despite strong evidence, published over three decades, the most commonly used strategies have strong evidence that demonstrate they are ineffective. There is a growing body of evidence to support newer interventions that are effective or show promise in reducing musculoskeletal pain and injuries in care providers. The authors have organized potential solutions into three established ergonomic solution types: engineering based, administrative, and behavioral. For each intervention, the level of evidence to support its use is provided. PMID- 17130759 TI - Update on evidence-based practices for safe patient handling and movement. PMID- 17130760 TI - NIOSH research efforts to prevent musculoskeletal disorders in the healthcare industry. AB - Healthcare workers, including orthopaedic nurses, face a number of risk factors in the workplace for musculoskeletal disorders such as back and shoulder injuries. These disorders are associated with excessive back and shoulder loading due to manual patient handling, applying excessive forces during pushing and/or pulling of objects, required use of awkward postures during patient care, and working long hours and shiftwork. No healthcare workers are immune from injury because workers in all clinical areas are exposed to occupational risk factors, including hospitals, nursing homes, emergency services, critical care, operating rooms, orthopaedic units, and home healthcare environments. This article includes a summary of the scientific efforts of the researchers and their partners at the National Institute for Occupational Safety and Health (NIOSH) in evaluating and developing the best practice recommendations for reducing risk of these disorders for exposed workers. The studies conducted by NIOSH researchers and their partners approach the problem from a variety of perspectives, ranging from comprehensive epidemiological studies examining the effectiveness of implementation of a safe patient handling and movement program to laboratory studies evaluating the biomechanical stress associated with using patient handling equipment, and education training programs for use in schools of nursing to educate new workers about safe work practices. Results of these studies have provided scientific evidence that significant occupational risks for musculoskeletal disorders exist and that effective interventions are available to reduce the risk for these workers. PMID- 17130761 TI - Broken bones: common pediatric lower extremity fractures--Part III. AB - Lower extremity injuries and fractures occur frequently in young children and adolescents. Nurses are often one of the first healthcare providers to assess a child with an injury or fracture. Although basic fracture care and principles can be applied, nurses caring for these young patients must have a good understanding of normal bone growth and development as well as common mechanisms of injury and fracture patterns seen in children. Similar to many of the injuries in the upper extremity, fractures in the lower extremity in children often can be treated nonoperatively with closed reduction and casting. However, this article will also review several lower extremity fractures that frequently require surgical intervention to obtain a precise anatomical reduction. Common mechanisms of injury, fracture patterns, and current management techniques will be discussed. Teaching strategies and guidelines that will enable nurses and nurse practitioners to confidently educate parents, families, and other providers caring for these young patients will be reviewed. PMID- 17130763 TI - Complications. AB - In orthopaedics, there are many potential complications that may either be life threatening or limb-threatening. Fat embolism, pulmonary embolism, and even hemorrhage may be life-threatening. Acute compartment syndrome, infection, and malunion or nonunion of a fracture can lead to loss of a limb or impaired limb functioning. Pain, if unrelieved, may cause physical and psychosocial impairment and is a priority concern from the patient's perspective. This guide is designed for the orthopaedic care provider to serve as a quick reference when reviewing these complications. It may be used as an educational tool in a handout or an enlarged poster format as a reminder of what to assess, what diagnostic tests to anticipate, and what interventions are necessary in these situations. Promotion of understanding of orthopaedic complications is important for their prevention, early recognition, and effective treatment to promote positive patient outcomes. PMID- 17130765 TI - An overview of radiography, computed tomography, and magnetic resonance imaging in the diagnosis of lumbar spine pathology. AB - When patients present with symptoms associated with lumbar spine pathology, often a series of diagnostic examinations of escalating sophistication are utilized. To obtain a diagnosis, the initial study is usually done on lumbar spine radiographs, which demonstrate gross bony pathologies, spinal alignment, and bone density. Frequently, additional high-cost invasive or noninvasive procedures may be required. Myelography is used to examine the spinal cord, nerve root bundles, and possible intrusion of the vertebral disk into the spinal canal. Computed tomography is most useful for imaging small bony structures and, when coupled with myelography, can demonstrate soft tissue abnormalities in the spinal canal. Magnetic resonance imaging is, however, the preferred modality for imaging soft tissue. PMID- 17130767 TI - Genetic testing and counseling: selected ethical issues. AB - As science and technology in genetics continue to expand, complex ethical questions arise that require difficult decisions for all concerned. Rather than having ready-made answers, there may be only more challenging questions for patients, families, and healthcare professionals. These complicated questions may have no straightforward, correct answers, thus creating an ethical quandary. This article discusses some selected ethical issues that occur when patients are considering undergoing genetic testing and counseling, including respecting autonomy and the patient's right to decide, establishing patient-healthcare provider trust, and disclosing private information to others. Several strategies are presented that nurses can use as they assist patients and their families who are facing these challenging ethical dilemmas. PMID- 17130770 TI - Registry of randomized controlled trials in transplantation: July 1-December 31, 2005 [corrected]. PMID- 17130773 TI - Designer genes: Filling the gap in transplantation. AB - Accelerated evolution of the field of functional genomics has been greatly facilitated by high-throughput microarray-based gene function studies, relating to the parallel and serial expression measurements of genomes. Microarray experimentation is being applied for the study of basic research questions, drug target discovery, pharmacology, toxicogenomics, target selectivity, disease biomarker determination, development of prognostic tests, and disease subclass determination. This article will review the current applications of microarray technology in the field of organ transplantation and discuss the potential impact of this technology on transplantation medicine. PMID- 17130774 TI - Marginal lung donors: A diminishing margin of safety? AB - Lung donor shortages have resulted in the critical appraisal of cadaveric donor acceptability criteria and the gradual relaxation of once strict guidelines. Many centers have reported their results with these "extended criteria" donors and an increasing number of multicenter registry studies have also been published. The results have been contradictory and leave many questions unanswered. Important new data has however come to light since the last review of the subject by the International Society for Heart and Lung Transplantation Pulmonary Council. We review the current literature focusing on recent developments in the pursuit of an expanded lung donor pool with acceptable outcomes. PMID- 17130775 TI - Pharmacodynamic monitoring of cyclosporine a in renal allograft recipients shows a quantitative relationship between immunosuppression and the occurrence of recurrent infections and malignancies. AB - BACKGROUND: At present it is unclear which dose and consecutive blood levels of cyclosporine A (CsA) are optimal with respect to immunosuppressive efficacy and drug specific side effects at the level of individual patients. Several pharmacodynamic measures of CsA effects have been proposed, but have not become clinical routine yet. Besides the lack of practicability, the biological relevance of these assays has not been determined so far. METHODS: Residual expression of nuclear factor of activated T-cells (NFAT)-regulated genes two hours after drug intake was used as molecular pharmacodynamic marker to assess CsA effects on lymphocytes and correlated with the frequency of recurrent infections and malignancies in patients with five or more years of follow-up posttransplantation. RESULTS: Recurrent infectious complications were observed in 44% and malignancies in 20% of the 133 patients studied. Patients with a strong suppression of NFAT-regulated genes by CsA--as judged by a residual level of transcription of less than 15% after drug intake--develop more frequent infections (53% vs. 29%; P = 0.005) and malignancies (22% vs. 4%; P = 0.002). The lack of correlation between the incidence of these complications and CsA blood concentration might point to the interindividual differences in the sensitivity towards calcineurin inhibition. CONCLUSION: The data presented here reveal a clear relation between the frequency of infectious and malignant complications and the degree of suppression of NFAT-regulated genes by CsA in transplanted patients. Therefore, pharmacodynamic monitoring of CsA efficacy in transplanted patients might be a useful tool to adjust immunosuppressive therapy in individual patients. PMID- 17130776 TI - Islet isolation and transplantation outcomes of pancreas preserved with University of Wisconsin solution versus two-layer method using preoxygenated perfluorocarbon. AB - BACKGROUND: Previous small clinical trials indicate that the two-layer method (TLM) for pancreas preservation improves islet isolation outcome. However, the effect of TLM has not been evaluated in large-scale study. In addition, a direct benefit of TLM on islet transplantation outcome has not been addressed in the setting of any randomized controlled trials. METHODS: Between April 2003 and October 2005, human pancreata from brain-dead donors were preserved by TLM using preoxygenated perfluorocarbon (n = 75) or in University of Wisconsin (UW) solution (n = 91) prior to islet isolation. Islet isolation and transplantation outcomes were compared between the two groups. RESULTS: We did not find any significant differences in adenosine triphosphate content in pancreatic tissue after preservation, pre and postpurification islet yields, in vitro insulin secretory function, or utilization ratio of transplantation between the two groups. Transplanted mass and functional viability of islet isolated from TLM preserved pancreas were similar to those from UW-preserved pancreas. Patients receiving the TLM-islet or the UW-islet showed a marked decrease in insulin requirement after transplantation. However, no significant difference was observed in a decrease in insulin requirement between patients receiving the TLM islet and the UW-islet. CONCLUSIONS: No beneficial effect of TLM on islet isolation and transplantation outcomes was observed. Our findings bring into question the true merit of routine use of TLM prior to islet isolation. PMID- 17130777 TI - Psychosocial and physical impairment after mini-incision open and laparoscopic donor nephrectomy: A prospective study. AB - BACKGROUND: The aim of the present study was to prospectively investigate how mini-incision donor nephrectomy (MIDN) and laparoscopic donor nephrectomy (LDN) affected the donor's quality of life and fatigue. METHODS: Forty-five donors underwent MIDN and 55 donors underwent LDN. Quality of life and fatigue were recorded preoperatively and four times during one year follow-up on the Short Form 36 (SF-36) and Multidimensional Fatigue Inventory-20 (MFI-20), respectively. RESULTS: One-year response rates were 89% and 95% following MIDN and LDN, respectively. After MIDN, all dimensions of the SF-36 significantly declined. Most dimensions returned to preoperative values at three months except for "vitality" (six months) and "bodily pain" (12 months). After LDN, the scores of the SF-36 dimensions returned to preoperative values at three months, except for "vitality" and "role physical" (both six months). Between-groups analysis revealed significantly better scores of the SF-36 dimensions "physical function" (P = 0.03) and "bodily pain" (P = 0.04) following LDN at one month postoperatively. Fatigue scores did not significantly differ between the groups at any point in time. General and physical fatigue (MFI-20) remained affected up to one year after either type of surgery. After MIDN, 4% of the donors had returned to work at four weeks postoperatively versus 28% after LDN (P = 0.04). Return to preoperative activity level was not significantly different between groups. CONCLUSIONS: Both procedures clearly impact quality of life and fatigue. The beneficial effect on the quality of life and the earlier return to work encourage us to advocate LDN as the surgical approach to be preferred. PMID- 17130778 TI - Comparison of renal allograft outcomes in combined liver-kidney transplantation versus subsequent kidney transplantation in liver transplant recipients: Analysis of UNOS Database. AB - BACKGROUND: There may be an allograft-enhancing effect by the liver on the renal allograft in the setting of simultaneous combined liver-kidney transplantation (CLKT) from the same donor. This study was performed to investigate whether an existing liver allograft could protect a kidney allograft from immunologic injury due to histoincompatibility in liver transplant recipients who received sequential kidney transplantation (KALT). METHODS: Using the United Network for Organ Sharing database covering January 1996 to December 2003, outcomes of 352 KALT were compared to 1,136 CLKT. Incidence of acute and chronic rejection and rejection-free renal graft survival was compared between two groups. RESULTS: Renal half-life of KALT allografts was shorter than CLKT group (6.6+/-0.9 vs. 11.7+/-1.3 years, P < 0.001). Incidence of chronic rejection in KALT group was higher than CLKT group (4.6 vs. 1.2%, P < 0.001). One and three-year rejection free renal graft survival of KALT and CLKT groups were different (77% and 67% KALT vs. 85% and 78% CLKT, respectively; P < 0.001). Among human leukocyte antigen mismatched and sensitized patients, rejection-free renal graft survival of KALT group was inferior to the CLKT group (75% at 1 year and 61% 3 years vs. 86% at 1 year and 79% 3 years, P < 0.001). CONCLUSION: Liver allograft provided renal graft immunoprotection if both organs are transplanted simultaneously (immunogenetic identity), but not for kidneys transplanted subsequently. PMID- 17130779 TI - Arginine and urea metabolism in the liver graft: A study using microdialysis in human orthotopic liver transplantation. AB - BACKGROUND: Arginine is an amino acid having a central role in the metabolism of urea and nitric oxide in the liver. We present our findings of the behavior of these metabolites during the process of transplantation of the liver. METHODS: Urea, arginine, ornithine, citrulline, gamma-aminobutyric acid, glutamate, and glutamine levels in 15 livers were studied during the process of retrieval, following storage during the backtable procedure, and for 48 hours postreperfusion using microdialysis. Arginase levels in donor and recipient serum were also analyzed using an enzyme-linked immunosorbent assay specific for type I human arginase. Data was analyzed using one-way analysis of variance, with post hoc comparison to the value at two hours using Dunnett's test (P < 0.05 significant). RESULTS: Levels of metabolites measured in the donor liver were seen to decline significantly in the stored liver. Immediately postreperfusion, there was a significant rise in arginase I levels in the recipient serum with low arginine levels recorded in the liver. The high arginase I levels significantly reduced six hours postreperfusion with a corresponding rise in extracellular arginine levels. Urea levels in the graft increased significantly immediately postreperfusion. CONCLUSIONS: Arginine levels were found to be low with correspondingly high serum arginase I levels in the early postreperfusion phase. High serum arginase I levels in early postreperfusion may influence nitric oxide production in this phase since considering Vmax and Km values, arginase I could compete with inducible nitric oxide synthase for arginine. Urea metabolism in the liver recommences immediately postreperfusion. PMID- 17130780 TI - Porcine model of auxiliary partial orthotopic liver transplantation for acute liver failure. AB - BACKGROUND: Auxiliary partial orthotopic liver transplantation (APOLT) has been an effective alternative in acute liver failure (ALF), but clinically several problems remain to be resolved. Thus, we attempt to establish an APOLT model for ALF using a large animal and demonstrate the validity of our model. METHODS: In experiment 1, we created an animal model of ALF using pig. ALF was induced by resection of 70% of the whole liver under total hepatic vascular exclusion (THVE). The duration of ischemia was 90 minutes. In experiment 2, we tried to make an APOLT model by using this ALF model as a recipient. That is, during 90 minutes of THVE, 70% hepatectomy and subsequent partial orthotopic transplantation was completed. RESULTS: In experiment 1, six of seven pigs died within three days with jaundice and massive ascites. Based on microcirculatory disturbance of the remnant liver and hepatocellular necrosis, 70% hepatectomy with 90 minutes of THVE was considered a proper model of ALF. In experiment 2, six out of seven APOLT model animals survived more than four days. T. Bil levels in the APOLT model remained consistently within the normal range throughout the observation period. In immunohistochemistry, several labeled nuclei stained with Ki67 were identified in native liver of the APOLT model. CONCLUSIONS: This APOLT procedure provided temporary liver function support and enabled the recipient to survive until the failing native liver had regenerated. Our APOLT model could be suitable and useful for understanding the role of APOLT in ALF. PMID- 17130781 TI - Liver repopulation by transplanted hepatocytes and risk of hepatocellular carcinoma. AB - BACKGROUND: Transplantation of isolated hepatocytes in rats treated with retrorsine (RS) results in massive repopulation of the host liver. In this study, the long-term fate of hepatocytes transplanted into RS-treated recipients was followed for up to two years. METHODS: Dipeptidyl-peptidase type IV-deficient (DPPIV) Fischer 344 rats were given two injections of RS (30 mg/kg), followed by transplantation of 2 million hepatocytes, isolated from a syngenic, DPPIV donor. RESULTS: Extensive (91+/-7%) liver replacement by transplanted hepatocytes was observed in animals sacrificed 18 months posttransplantation. Similar levels of repopulation persisted at two years (87+/-5%). No evidence of preneoplastic and/or neoplastic evolution of the transplanted cell population was present in the RS-treated and repopulated livers at any time point considered. Furthermore, serum parameters related to hepatocyte function and integrity were in the normal range. In control groups given cell transplantation in the absence of prior treatment with RS, only small clusters of donor-derived, DPPIV hepatocytes were discerned. CONCLUSIONS: These results indicate that liver repopulation in this model is largely stable, persisting for up to two years and allowing for a normal liver function. In addition, no increased risk of neoplastic transformation appears to be associated with the process of liver repopulation for as long as over two thirds of the life span of the recipient animal. PMID- 17130782 TI - Interleukin-12p70 prolongs allograft survival by induction of interferon gamma and nitric oxide production. AB - BACKGROUND: Interleukin (IL)-12p70, a heterodimeric cytokine has been considered central to induction of Th1 responses with the assistance of IL-18 and IL-27. It was predicted IL-12p70 treatment would promote allograft rejection. In these studies, IL-12p70 delayed rejection. METHODS: We compared Piebald Virol Glaxo (PVG) neonatal heart graft survival in fully allogeneic Dark Agoutti (DA) rats treated with IL-12p70 alone or in combination with other cytokines. The mechanism by which IL-12p70 induced delayed rejection was examined by reverse transcription polymerase chain reaction of cytokine mRNA and studying the role of interferon (IFN)-gamma and inducible nitric oxide synthase (iNOS) that were induced by IL 12. RESULTS: IL-12p70 treatment significantly delayed PVG neonatal heart graft rejection compared to normal rejection control and other control groups treated with supernatant from Chinese hamster ovary (CHO)-K1 cells transfected with IL 12p35, IL-12p40, or no cytokine gene. IL-12p70 had no effect on alloantibody response. IFN-gamma and iNOS mRNA expression was increased in heart graft and regional lymph node compared to normal rejection and other treatment groups, consistent with Th1 response induction. IL-12p35 mRNA expression decreased in IL 12p70 treated rats but there was no difference in IL-12p40, Th2, or Tr1 cytokine mRNA expression. Coadministration of an iNOS inhibitor, L-NIL, or a monoclonal antibody (mAb) that blocks IFN-gamma, inhibited IL-12p70's ability to prolong allograft survival; as did co-treatment with IL-4 but not IL-13. CONCLUSIONS: IL 12p70 treatment may inhibit rejection by hyperinduction of Th1 responses, especially production of IFN-gamma and nitric oxide. These effects may be by enhancing regulatory T-cell responses or by the activation of iNOS in macrophages to produce excessive nitric oxide that in turn inhibits alloimmune responses. PMID- 17130783 TI - A negative regulatory role in mouse cardiac transplantation for a splice variant of CD80. AB - BACKGROUND: Members of the B7 costimulatory protein family (CD80 and CD86) play a determining role in allograft rejection. Both CD80 and CD86 have naturally occurring splice variants whose roles in transplantation are unknown. Full length CD80 has two immunoglobulin (Ig)-like domains in the extracellular portion, IgC and IgV. In mouse, the isoform IgV-CD80 lacks the IgC-like domain. Here we analyzed the role of mouse IgV-CD80 in heart allograft rejection and search for equivalent splice variants in human. METHODS: Mice made deficient for full-length CD80 but which retain expression of the shorter IgV-CD80 (CD80 mice) were used as donor or recipient of a heart allograft. Recipient animals were untreated or pretreated with alloantigen expressing cells and/or treated with CD80 and CTLA4 monoclonal antibodies (mAbs). RESULTS: Recipients expressing IgV-CD80 but not full length CD80 exhibited a slight prolongation in survival of either wild-type (Wt) or CD80 grafts. More dramatically, CD80 animals pretreated with donor alloantigen exhibited permanent graft survival, whereas their Wt counterparts rejected their grafts with a median survival of 24 days. This prolonged survival was due to the expression of IgV-CD80 in recipients since treatment with CD80 mAb abrogated the beneficial effect observed. We identified and report here a similar isoform of CD80 from human cDNA encoding a putative soluble, IgV-containing protein. CONCLUSIONS: IgV-CD80 bearing recipients show enhanced allograft survival especially after donor alloantigen pretreatment. This together with data from other species suggests that regulation delivered by splice variants of CD80 significantly modulates immunity and may be common across the species. PMID- 17130784 TI - Recipient memory-like lymphocytes remain unresponsive to graft antigens after CAMPATH-1H induction with reduced maintenance immunosuppression. AB - BACKGROUND: Treatment with CAMPATH-1H at the time of transplantation allows reduced maintenance immunosuppression. We hypothesized that CAMPATH-1H induction would modulate the response of repopulating leukocytes to donor alloantigens. METHODS: The phenotype, proliferative and stimulatory capacity of peripheral blood leukocytes from 14 renal transplant recipients treated with CAMPATH-1H and reduced immunosuppression with mycophenolate mofetil and tacrolimus were investigated for the first six months after transplantation. The impact of immunosuppressive drugs on leukocytes that escape depletion was also evaluated in vitro. RESULTS: CAMPATH-1H therapy caused a significant decrease in the number of B and T cells, with CD4 T central memory cells being the most resistant to depletion. The recovery of CD8 T cells was faster than that of CD4 T cells. Lymphopenia correlated with a decrease in both proliferative and effector responses, however, the recipient T cells remained responsive to third-party antigens. Dendritic cells (DC) were also depleted but to a lesser extent than lymphocytes; lymphoid DC were more resistant than myeloid DC; these changes correlated with decreased allostimulatory capacity. One of the patients experienced rejection that was treated successfully. The rejection was associated with a high proportion of CD4 T effector memory cells and myeloid DC, increased proliferation and enhanced effector activity to donor antigens. In vitro studies confirmed that the reduced dose of immunosuppressive drugs used could prevent activated T cells from switching to the effector compartment, suppressing both their proliferation and effector activity. CONCLUSIONS: CAMPATH-1H induction combined with reduced maintenance immunosuppression is sufficient to control the effector phase of immune response to donor antigens. PMID- 17130785 TI - Controlling the generation and function of human CD8+ memory T cells in vitro with immunosuppressants. AB - BACKGROUND: Memory T cells play a pivotal role in acute and chronic rejection of transplanted organs. Novel therapies such as alemtuzumab and the identification of heterologous memory have highlighted their importance following transplantation. Unlike naive T cells, the influence of immunosuppressive agents on the de novo generation of memory T cells and on the function of pre-existing memory T cells is poorly understood. METHOD: CD8 effector memory T cells (Tem, CD45RA, CCR7) were generated by stimulating peripheral blood mononuclear cells for five days with anti-CD3 and anti-CD28 monoclonal antibodies, followed by nine days of rest. The influence of immunosuppression on this was assessed by flow cytometry. Tem that had been naturally formed in vivo were used to study the influence of the same agents on the function (intracellular interferon-gamma production) of Tem cells. RESULTS: Whereas all the immunosuppressive agents tested inhibited the expansion of CD8 Tem population by influencing their proliferation and apoptosis, the calcineurin inhibitors were better at controlling their function. Furthermore, Tem generated despite the presence of tacrolimus were functional and did not exhibit drug resistance. CONCLUSIONS: Immunosuppressive agents differ in their ability to control the generation and function of CD8 Tem; the calcineurin inhibitors being the most efficacious. This may be of importance when tailoring immunosuppressants for transplant recipients, particularly in those where there is pre-existing immunological memory or in those receiving T-cell depleting therapies, which skews the immune repertoire towards memory. PMID- 17130786 TI - Cardiac and skin xenograft survival in different recipient mouse strains. AB - BACKGROUND: There are conflicting reports on the importance of antibody and cell mediated mechanisms and the influence of TH1 or TH2 cytokines on acute vascular xenograft rejection. We sought to resolve some of the recent discrepancies in the rat-to-mouse xenograft model where different recipient strains are used and investigated the TH1/TH2 influence on rejection. METHODS: Lewis rat heart xenograft survival was compared between BALB/c and C57BL/6 recipients. Antigraft antibody deposition, serum anti-rat antibody levels and B-cell deficient recipients were used to examine the contribution of antibody to rejection. To further investigate a TH1 or TH2 bias effect in vivo, we used BALB/c STAT4 knockout (KO) and STAT6 KO recipient mice. Experiments were repeated with rat skin xenografts to examine TH1/TH2 influences on cell-mediated rejection. RESULTS: The median survival (MS) of rat heart xenografts in BALB/c and C57BL/6 mice was five and eight days, respectively (P = 0.002). The MS in B-cell deficient mice was 16 days (P < 0.001). The MS in STAT4 KO and STAT6 KO mice was six and seven days respectively (P = 0.009). All non-B-cell deficient recipients showed strong IgM deposition and histological features of both cellular and antibody-mediated rejection. There was no correlation between serum anti-rat antibody levels and graft outcome or graft deposition. There was no survival difference of skin xenografts in BALB/c, C57BL/6, B-cell deficient, STAT6 KO, or STAT4 KO mice (8-9 days). CONCLUSIONS: Both humoral and cell-mediated immunity have significant roles in vascularized heart xenograft rejection. TH1/TH2 biases minimally affect rejection through humoral but not cellular immunity. PMID- 17130787 TI - Sensitive balance of suppressing and activating effects of mesenchymal stem cells on T-cell proliferation. AB - The human leukocyte antigen-independent immune-modulatory potential of human mesenchymal stem cells (hMSC) makes them a promising candidate for clinical cell therapy. A better understanding of their "immune-privileged" status is therefore of high priority. Here we used Ki-67-antigen staining to estimate T-cell alloreactivity in mixed lymphocyte cultures in the presence of hMSC as second or third party. We found that the allostimulatory activity of mesenchymal stem cells (MSC) leading to an increased T-cell proliferation and interleukin-2 secretion is measurable only at low MSC/effector ratios (< or =0.1:1). Moreover, this stimulating effect could be efficiently suppressed by MSC-conditioned medium. This suggests that the "immune-privileged" status of MSC exists only when MSC mediated downregulation of immune cell activation can overrule their own allostimulatory potential. Thus the "immune-privileged" state of MSC represents a sensitive balance of suppressing and activating effects, which should be considered in a clinical setting with limited cell amounts. PMID- 17130788 TI - Impact of mismatching CD1a, a dimorphic antigen-presenting molecule, on graft versus-host disease after hematopoietic stem cell transplantation. AB - CD1a, an antigen-presenting molecule related to major histocompatibility complex (MHC) class I, is frequently described as nonpolymorphic. In humans it is dimorphic, due to two linked amino acid substitutions in the alpha1 domain (Ile13Thr and Trp51Cys). The CD1a gene on chromosome 1 is not linked to MHC and may be mismatched between human leukocyte antigen-identical siblings. We analyzed 155 donor-recipient pairs of the Eurobank cohort, 141 matched for CD1a and 14 unmatched in the graft-versus-host disease (GVHD) direction. The burden of GVHD was not increased by CD1a mismatching. The incidence of GVHD in matched and unmatched groups was respectively: grade I-IV: 81% and 86% (P = 0.492); II-IV 61% and 57% (P = 0.495); III-IV 23% and 21% (P = 0.608). Adjusting for age, sex mismatch, GVHD prophylaxis, and conditioning did not reveal any significant difference. This suggests that, unlike conventional class I molecules, CD1a does not function as a transplantation antigen and does not require matching in hematopoietic stem cell transplantation. PMID- 17130789 TI - Inhibition of apoptosis reduces immunogeneic potential of adenoviral-treated syngeneic liver grafts. AB - Effects of adenoviral therapy and reduced apoptosis on immune response were investigated in a rat liver transplantation model after prolonged ischemia reperfusion. Liver donors were treated i.v. either with an adenoviral construct, expressing bcl-2, green-fluorescent-protein, or doxycyclin. Intrahepatic apoptosis was assessed by terminal transferase dUTP nick end labeling assay. The intrahepatic presence of CD4, CD8a, CD163, immunoglobulin (Ig)beta, tumor necrosis factor (TNF)-alpha and myeloperoxidase (MPO) was quantified by realtime polymerase chain reaction at 24 hours and seven days after transplantation. Bcl-2 expression abrogated the TNF-alpha elevation and reduced apoptosis of hepatocytes and sinusoidal endothelial cells as compared to advCMV green fluorescent protein. No effects on CD4, CD8a, CD163 and MPO expression were noticed in bcl-2 pretreated livers, whereas Igbeta was slightly enhanced compared to controls. Adenoviral infected liver grafts trigger an immune response but reduced apoptosis resulted in down-regulation of TNF-alpha. Thus, bcl-2 transfer might simultaneously reduce graft ischemia reperfusion injury and immunogenicity. PMID- 17130790 TI - Toll-like receptor-2 Arg753Gln polymorphism and herpes virus infections in transplantation. PMID- 17130791 TI - Need for a registry of living kidney donor outcomes. PMID- 17130792 TI - Antifungal alternatives for invasive candidiasis refractory to caspofungin in liver transplant recipients: A report of two cases. PMID- 17130793 TI - Mycophenolate mofetil and bronchiectasis in pediatric transplant patients. PMID- 17130794 TI - Standards for ethical publication. PMID- 17130796 TI - Tinnitus sensitization: Sensory and psychophysiological aspects of a new pathway of acquired centralization of chronic tinnitus. AB - OBJECTIVE: Acquired centralized tinnitus (ACT) is the most frequent form of chronic tinnitus. The proposed ACT sensitization (ACTS) assumes a peripheral initiation of tinnitus whereby sensitizing signals from the auditory system establish new neuronal connections in the brain. Consequently, permanent neurophysiological malfunction within the information-processing modules results. Successful treatment has to target these malfunctioning information processing. We present in this study the neurophysiological and psychophysiological aspects of a recently suggested neurophysiological model, which may explain the symptoms caused by central cognitive tinnitus sensitization. Although conditioned reflexes, as a causal agent of chronic tinnitus, respond to extinction procedures, sensitization may initiate a vicious circle of overexcitation of the auditory system, resisting extinction and habituation. DATA SOURCES: We used the literature database as indicated under "References" covering English and German works. STUDY SELECTION: For the ACTS model we extracted neurophysiological hypotheses of the auditory stimulus processing and the neuronal connections of the central auditory system with other brain regions to explain the malfunctions of auditory information processing. The model does not assume information processing changes specific for tinnitus but treats the processing of tinnitus signals comparable with the processing of other external stimuli. The model uses the extensive knowledge available on sensitization of perception and memory processes and highlights the similarities of tinnitus with central neuropathic pain. DATA EXTRACTION: Quality, validity, and comparability of the extracted data were evaluated by peer reviewing. DATA SYNTHESIS: Statistical techniques were not used. CONCLUSION: According to the tinnitus sensitization model, a tinnitus signal originates (as a type I-IV tinnitus) in the cochlea. In the brain, concerned with perception and cognition, the 1) conditioned associations, as postulated by the tinnitus model of Jastreboff, and the 2) unconditioned sensitized stimulus responses, as postulated in the present ACTS model, are actively connected with and attributed to the tinnitus signal. Attention to the tinnitus constitutes a typical undesired sensitized response. Some of the tinnitus-associated attributes may be called essential, unconditioned sensitization attributes. By a process called facilitation, the tinnitus' essential attributes are suggested to activate the tinnitus response. The result is an undesired increase in responsivity, such as an increase in attentional focus to the eliciting tinnitus stimulus. The mechanisms underlying sensitization are known as a specific nonassociative learning process producing a structural fixation of long-term facilitation at the synaptic level. This sensitization model may be important for the development of a sensitization-specific treatment if extinction procedures alone do not lead to satisfactory outcome. Inasmuch as this model considers sensitization as a nonassociative learning process based on cortical plasticity, it is reasonable to assume that this learning process can be altered by counteracting learning procedures. These counteracting learning procedures may consist of tinnitus-specific cognitive and behavioral procedures. PMID- 17130797 TI - Silicone allergy: A new cause for cochlear implant extrusion and its management. AB - OBJECTIVE: We introduce silicone allergy as a rare cause for cochlear implant extrusion and discuss its management. STUDY DESIGN: Retrospective case series and literature review. SETTING: Tertiary referral centers. PATIENTS: Primary eligibility criteria included patients who experienced a delayed extrusion of their cochlear implants with negative wound cultures and had a suspected or a test-proven allergy to silicone components of an implant. INTERVENTIONS: Silicone allergy testing, explantation of a cochlear implant containing allergenic silicone materials, reimplantation with a custom-made cochlear implant excluding an allergenic silicone component. OUTCOME MEASURES: Uneventful wound healing and extrusion-free long-term follow-up after the reimplantation with a custom-made cochlear implant excluding an allergenic silicone component. RESULTS: Three known cases of cochlear implant extrusion as a result of silicone allergy have been noted from 1991 through 2004 in three cochlear implant programs in the United States. All three devices extruded, resulting in explantation of the old device and reimplantation with a new custom-made device eliminating the allergenic silicone component. Wound cultures were negative in all cases. All three patients experienced a delayed extrusion of their devices. Two of these patients had a test-proven allergy to the implant's silicone components, whereas the third patient was presumed to have a hypersensitivity solely on the basis of a clinical presentation. CONCLUSION: We propose that silicone allergy is a rare cause of cochlear implant extrusion. Patients experiencing cochlear implant extrusion, particularly with a delayed onset and negative wound culture results, should be tested for silicone allergy. PMID- 17130798 TI - Conservation of residual acoustic hearing after cochlear implantation. AB - OBJECTIVE: This study was designed to test the hypothesis that partial hearing conservation is attainable after cochlear implantation with a long perimodiolar electrode. Surgical strategies for hearing conservation during cochlear implantation are described. STUDY DESIGN: Prospective, single-subject, repeated measures design. SETTING: Academic tertiary care center. PATIENTS: Twenty-eight severely to profoundly hearing-impaired adult cochlear implant recipients who had some measurable hearing preoperatively. INTERVENTION: Cochlear implantation using Nucleus Freedom Contour Advance electrode. MAIN OUTCOME MEASURES: Preimplant and postimplant pure-tone thresholds and speech recognition scores were obtained to determine the incidence and degree of conserved hearing at a mean interval of 9 (+/-3.9) months. RESULTS: Thirty-two percent of subjects experienced complete conservation of hearing (0- to 10-dB loss), and 57% experienced partial conservation of hearing (>11 dB) after implantation. However, open-set speech recognition was partially conserved in only one subject. Cochlear implant performance was not better in patients with conservation of residual hearing. CONCLUSION: Conservation of pure-tone hearing was possible in 89% of implanted patients; however, residual speech perception was not conserved with this long perimodiolar electrode. A ceiling effect tends to inflate the prevalence of hearing conservation in implantation studies of severely to profoundly hearing impaired patients. PMID- 17130799 TI - Management of lateral sinus thrombosis in chronic otitis media. AB - OBJECTIVE: To present our experience with the diagnosis and treatment of lateral sinus thrombosis (LST) secondary to otogenic disease. PATIENTS AND METHODS: We have retrospectively reviewed all the cases of LST caused by chronic otitis media in ten years. RESULTS: In our series of five patients, four patients were successfully treated by surgery without opening the sigmoid sinus. On the other hand, one of our patients developed hydrocephalus and ataxia during preoperative antibiotic treatment. CONCLUSION: Most patients with thrombotic/nonabscessed sinuses will have successful outcomes with this approach, but careful observation is needed, given the possibility of advancing disease. PMID- 17130800 TI - Study of vestibular evoked myogenic potentials in unilateral vestibulopathy: Otolithic versus canal function testing. AB - OBJECTIVE: The study provides a qualitative evaluation of unilateral vestibulopathy by comparing otolithic and canal function, to establish possible relationships between the type of dysfunction observed and the evolving clinical pictures associated with it. STUDY DESIGN: Retrospective study of a series of cases. SETTING: Department of Medical-Surgical Specialization, Otolaryngology and Cervicofacial Surgery Division, University of Perugia, Perugia, Italy. PATIENTS: Twenty patients whose medical history showed at least one episode corresponding to the clinical parameters of acute vestibulopathy. INTERVENTIONS: Study of vestibular function by recording VEMPs and repeating canal function testing at least 6 months after the first episode of vertigo. MAIN OUTCOME MEASURES: Relationship between the type of vestibulopathy (canal and otolithic) and the clinical pictures observed. RESULTS: Paroxysmal positional vertigo, observed in 4 patients, was correlated with the presence of vestibular evoked myogenic potentials (VEMPs) and the absence of an ipsilateral canal response in all cases (100%). Persistent dizziness was observed in nine patients, and VEMPs were absent in all of them (100%); three (33.3%) showed the recovery of previously absent canal function. Comparison of responses in six patients with recurrent acute vestibulopathy showed persistent and complete loss of canal function in five cases (83.3%), whereas impairment of otolithic response was less constant (40%). CONCLUSION: The combined VEMPs-canal test study shows predictive value regarding certain evolving clinical pictures of vestibulopathy. The absence of VEMPs confirms the role of otolithic dysfunction in the onset of dizziness. Likewise, it suggests that a vestibular origin of these disorders should be considered in cases that have shown aspecific symptoms since onset, without frank vertigo and with normal vestibular response to canal function testing. PMID- 17130801 TI - Alternobaric vertigo--really a hazard? AB - OBJECTIVE: To determine the prevalence of alternobaric vertigo (AV) in sport divers and to find out whether AV led to dangerous situations underwater. Furthermore, to examine whether objective neurootologic tests are associated with the manifestation of AV. DESIGN: Retrospective cohort study. PARTICIPANTS: Sixty three sport divers with an average diving experience of 10 years and 650 dives were questioned regarding their medical and diving history and the manifestation of vertigo during diving. METHODS: Microscopic otoscopy, tympanometry, stapedius reflexes, hearing threshold for air and bone conduction, caloric video oculography including analysis of the slow-phase velocity of the nystagmus, acoustic brain stem responses, and magnetic resonance imaging were performed to find possible differences between divers with and without AV. RESULTS: We found 17 divers with AV (27%). There was no significant difference in all measured parameters apart from sex and history of middle ear equalization difficulty in divers with AV. Ten (59%) of 17 female divers and 7 (15%) of 46 male divers experienced AV, representing a significant sex difference (p < 0.001). Correlation with our divers' outpatient clinic revealed that female divers had a significantly higher incidence of middle ear equalization disorders which could be an explanation for the predominance of female divers with symptoms of AV. None of the divers reported any dangerous or life-threatening situations following AV. Whether AV leads to dangerous situations underwater remains unclear, but this hypothesis is not supported by our data. CONCLUSION: Alternobaric vertigo is a common finding in divers. In our study group, female divers had a four-time higher risk to suffer AV. Our data do not support the thesis that AV is a life threatening condition. PMID- 17130802 TI - Preservation of conductive hearing in approaches to tumors of the jugular foramen. AB - OBJECTIVE: Surgical approaches to the jugular foramen, most often used for the resection of glomus jugulare tumors, may include removal of the external auditory canal wall and overclosure of the meatus, resulting in maximal conductive hearing loss. Modifications have been described that maintain hearing by preserving the canal wall at the price of decreased exposure and are, therefore, suitable only for small and favorably located tumors. Our technique for removal and then reconstruction of the canal wall with hydroxyapatite cement allows for complete anterior translocation of the facial nerve as far proximal as the geniculate ganglion, giving uncompromising exposure of even the most extensive tumors, with the potential for preservation of normal hearing. The purpose of this study was to describe and report our experience with this technique. STUDY DESIGN: : Retrospective review. SETTING: Private otology practice. PATIENTS: Between 2000 and 2005, seven patients between the ages of 34 and 77 years were identified who underwent procedures using this technique. INTERVENTION: Surgical management of jugular foramen tumors. MAIN OUTCOME MEASURES: Successful anatomical reconstruction of the external auditory canal and middle ear. Preoperative and postoperative audiograms are compared, and facial nerve function is reported. Complications are discussed. RESULTS: All seven patients had successful reconstruction of the external auditory canal. Complications were minor and did not require additional surgical intervention. Postoperative pure-tone average differed from the preoperative average by a mean of 7.5 dB. Facial nerve function ranged from House-Brackmann Grades I to III when checked at least 7 months after surgery. CONCLUSION: This study reveals that this technique of external auditory canal reconstruction using hydroxyapatite cement allows complete anterior translocation of the facial nerve, while safely and reliably preserving the potential for normal hearing, without any compromise in exposure of the jugular foramen in the setting of an infratemporal fossa approach. PMID- 17130803 TI - Intracranial schwannomas of the lower cranial nerves. AB - OBJECTIVE: To present our experience in the diagnosis and management of 39 patients with lower cranial nerve schwannomas of the posterior fossa. STUDY DESIGN: A retrospective chart review of patient medical records. SETTING: Tertiary care, academic medical center. PATIENTS: All patients with intracranial lower cranial nerve schwannomas treated surgically in our institution between July 1998 and July 2005. INTERVENTION: A retrosigmoid, transcondylar, or combined approach was used for tumor recurrence. RESULTS: Thirty-nine patients underwent surgical resection, with complete tumor removal in 32, near-total resection in 5 patients, and subtotal tumor excision in 2 patients. Long-term (mean, 8.2 years) magnetic resonance imaging surveillance demonstrated recurrent tumor in 2 of 32 complete resections and slow regrowth in 2 of 7 patients with known residual disease. Only one of these four patients required reoperation. DISCUSSION: Intracranial schwannomas of the lower cranial nerves are relatively uncommon and may present with subtle or no clinical symptoms. Successful surgical resection with low risk of tumor recurrence can be achieved with the retrosigmoid or transcondylar approach. Morbidity, in this series, was primarily related to lower cranial nerve deficits. PMID- 17130805 TI - Social and behavioral determinants of sexually transmitted disease: scientific and technologic advances, demography, and the global political economy. PMID- 17130806 TI - Syphilis returns to china ... With a vengeance. PMID- 17130807 TI - Editorial. PMID- 17130808 TI - Artificial beta-cell: clinical experience toward an implantable closed-loop insulin delivery system. AB - AIM: Restoration of long-term normal blood glucose control in diabetic patients supports the elaboration of an artificial beta cell. The possibility of implantation of the three crucial components of such a system (insulin delivery device, glucose sensor and controller) is analyzed. METHODS: The Long-Term Sensor System project, aiming at a fully implantable artificial beta cell, assessed the feasibility of glucose control by the combined implantation of a pump for peritoneal insulin delivery and a central intravenous glucose sensor close to the right atrium, connected via a subcutaneous lead. It was initiated in 10 Type 1 diabetic patients in our clinic from 2000. Data obtained during this experience are reviewed and confronted to reported closed-loop trials using other approaches. RESULTS: No significant complication related to prolonged implantation of intravenous sensors occurred and the combined implants were well tolerated. Glucose measurement by the intravenous sensors correlated well with meter values (r=0.83-0.93, with a mean absolute deviation of 16.5%) and accuracy has been sustained for an average duration of 9 months. Uploading of pump electronics by algorithms designed for closed-loop insulin delivery allowed in patient 48 hour-trials aiming at automated glucose control. Glucose control was similar to that reported by investigations combining subcutaneous sensors to wearable pumps for subcutaneous insulin infusion. The benefits of more physiological insulin kinetics due to intra-peritoneal delivery have been hampered by the slow response time of intravenous sensors. CONCLUSION: Although the concept of a fully implantable artificial beta cell has been validated as feasible, the limited performance in achieving glucose control requests improvements in the sensor structure to increase its longevity and decrease sensor delay. PMID- 17130809 TI - Monitoring of the islet graft. AB - The Edmonton trials have brought about a marked improvement in the short-term rate of success of islet transplantation with rates of insulin-independence of 80% at 1-year being reported by several institutions worldwide. Unfortunately, this rate consistently decreases to 10-15% by 5 years post-transplantation. Several mechanisms have been proposed to explain this apparent 'islet exhaustion', but are difficult to pinpoint in a given patient. Understanding the reasons for islet graft exhaustion and its kinetics is a prerequisite for the improvement of islet transplantation outcome. In this regard, efficient monitoring tools for the islet graft have been conspicuously lacking and are required to detect islet damage and diagnose its mechanisms in a timely fashion, so as to initiate salvage therapy such as antirejection treatment. Tools for the monitoring of the islet graft include follow-up of metabolic function but mostly indicate dysfunction when it is too late to take action. Progress is likely to arise in the fields of immune monitoring, molecular monitoring and islet imaging, notably thanks to magnetic resonance (MR) or positron emission tomography (PET) technologies. PMID- 17130810 TI - Alternatives routes of insulin delivery. AB - Optimal glycaemic control is necessary to prevent diabetes-related complications. An intensive treatment, which could mimic physiological insulin secretion, would be the best one. However subcutaneous insulin treatment is not physiologic and represents a heavy burden for patients with type 1 and type 2 diabetes mellitus. Consequently, more acceptable, at least as effective, alternative routes of insulin delivery have been developed over the past years. Up to now, only pulmonary administration of insulin (inhaled insulin) has become a feasible alternative to cover mealtime insulin requirements and one of the various administration systems was recently approved for clinical use in Europe and the United States. But, due to advances in technology, other routes, such as transdermal or oral (buccal and intestinal) insulin administration, could become feasible in a near future, and they could be combined together to offer non invasive, efficacious and more physiological way of insulin administration to patients with diabetes. PMID- 17130811 TI - Immunology of pancreatic islet transplantation. AB - Clinical protocols in type 1 diabetic patients to optimize islet survival and function post-transplantation improved dramatically in the last decade, but it is clear that this approach still has potential limitations to provide long term insulin independency. Islet allografts administered in the liver via the portal vein are exposed to several factors contributing to a rapid loss of function that may reach 50% of the initial beta cell mass. Allo- and auto-immune reactions - an unique situation in clinical transplantation - are partially overcome with immunosuppressive regimen. Serological markers and T cell reactivities may correlate with graft failure. Most of the drugs that are used, including rapamycin (sirolimus) or the calcineurin inhibitor tacrolimus (FK506), have deleterious effects on beta function and/or insulin sensitivity. Immediate factors that limit initial islet engraftment have been elucidated, including instant blood mediated inflammatory reaction and angiogenesis. Newer interventions designed to promote islet survival, to prevent apoptosis, to promote islet growth and to protect islets in the long run from immunological injury are rapidly approaching clinical trials. PMID- 17130812 TI - Innovative therapies: some ethical considerations. AB - The purpose of this text is to try to understand why certain innovations are not permitted, although they are possible. Our guiding thread will be the four principles of biomedical ethics defined by Beauchamp and Childress: beneficence, non-maleficence, justice and autonomy. We shall show how they can guide the ethical inquiry in the field of pancreas or islet transplantation, leading to an analysis of the risks and benefits of the innovations epistemologically taking into account their historical context. PMID- 17130813 TI - Development of the endocrine pancreas. AB - This short review presents the recent breakthroughts in our understanding of the important steps controlling pancreas morphogenesis and differentiation, and on the transcription factors regulating pancreas organogenesis and islet cell differentiation and involved in the specification of the beta and alpha cell lineages. All these studies should permit a comprehensive view of the full genetic program necessary to produce mature and functional beta cells and thus, should be instrumental to guide future strategies for cell replacement therapies in type 1 diabetes. PMID- 17130814 TI - Continuous glucose monitoring and external insulin pump: towards a subcutaneous closed loop. AB - The development of an artificial pancreas for the treatment of type 1 diabetes is a highly desired endeavour for type 1 diabetic patients, physicians, scientists and engineers. The development of the first miniaturized external pumps in the 70s and the pharmacokinetic properties of short acting insulin analogues, closer to physiology, have raised hopes for the elaboration of such a device. Recent technological progress in the development of continuous glucose sensors, have improved the reliability and accuracy of these devices. This has led to the development of prototypes of closed-loop system based on the combination of a continuous monitor, a control algorithm, and an insulin pump. This review focuses on the SC-SC approach, employing subcutaneous glucose monitoring and subcutaneous insulin delivery. The feasibility of this solution has been proven at a small scale, but remains to be confirmed in the home setting. Intermediate solutions, such as semi-automatic systems, might be immediately valuable. PMID- 17130815 TI - Immunosuppressive drug-induced diabetes. AB - Post-transplant diabetes mellitus (PTDM) has emerged as a major adverse effect of immunosuppressive drugs (ISD). As recipients of organ transplants survive longer, the complications of diabetes mellitus have assumed greater importance. The predominant factor for causing PTDM by corticosteroids seems to be the aggravation of insulin resistance, however several studies have displayed deleterious effects on insulin secretion and beta-cells. Calcineurin inhibitors induce PTDM by a number of mechanisms, including decreased insulin secretion and a direct toxic effect on the pancreatic beta-cells. Recent in vitro studies stress on the increased apoptosis of beta-cells when exposed to these drugs. Studies involving other immunosuppressive agents (mycophenolate mofetil [MMF], sirolimus) are scarcer and lead to conflicting results, while daclizumab seems to have a neutral effect. Clinical studies have consistently shown a greater potential of tacrolimus to induce PTDM compared with cyclosporine. Reducing PTDM incidence is a feasible goal while using corticosteroid-sparing regimens and/or lower tacrolimus trough levels. In patients developing PTDM, conversion from tacrolimus to cyclosporine could improve or reverse glucose tolerance abnormalities. In the absence of well-designed studies in this specific indication, treatment of PTDM is based on the same principles as type 2 diabetes mellitus. Thiazolidinediones do not display any pharmacological interaction with calcineurin inhibitors, but their safety and efficacy in PTDM need to be confirmed in large-scale randomized trials. Use of sulfonylureas has to be cautious regarding the suspected interaction of some of them with calcineurin inhibitors. If needed, insulin regimens have to be adapted in patients who display the particular glycaemic profile of corticosteroid-induced diabetes. Incretin-based therapies, due to their specific action on beta-cell apoptosis and proliferation, raise promises that have to be confirmed in clinical studies. Until methods for inducing specific graft tolerance become available, immunosuppressive regimens should be tailored to the individual patient on the basis of predictive criteria for the development of PTDM. PMID- 17130817 TI - Tumoral calcinosis in a peritoneal dialysis patient. PMID- 17130818 TI - Deferoxamine-related fatal nasal-orbital-cerebral mucormycosis. PMID- 17130819 TI - Nuclear receptors link gender dimorphism of renal disease progression. AB - Profound gender differences in the progression of chronic kidney disease (CKD) remain poorly understood. Differential expression of the genes for male- and female-specific proteins in the kidney has been proposed to account for this clinical phenomenon. Lu and colleagues provide evidence that the signaling pathways of the nuclear receptors ERalpha, AR and PPARalpha are associated with gender differences in CKD progression. PMID- 17130820 TI - 4-Hydroxyproline metabolism and glyoxylate production: A target for substrate depletion in primary hyperoxaluria? AB - The primary hyperoxalurias are diseases of overproduction of oxalate. The immediate precursor of oxalate is glyoxylate. Metabolism of hydroxyproline, derived from collagen turnover or the diet, appears to be a major source of glyoxylate, and a potential target for a therapeutic strategy of substrate depletion. PMID- 17130821 TI - The broader burden of end-stage renal disease on children and their families. AB - When a child has end-stage renal disease and requires dialysis, a heavy personal and financial toll can be extracted from the caregivers and the family. Tsai et al. have demonstrated an adverse effect on the psychosocial and socioeconomic well-being of caregivers of children on chronic peritoneal dialysis. These findings raise other questions and force us to think about support for the caregiver as well as the patient. PMID- 17130824 TI - Water-only pores and peritoneal dialysis. PMID- 17130826 TI - Could tubular interstitium be a source of adult epithelial stem cells? PMID- 17130827 TI - Sporadic human renal tumors display frequent allelic imbalances and novel mutations of the HRPT2 gene. AB - Inactivation of the HRPT2 gene encoding parafibromin was recently linked to the familial hyperparathyroidism-jaw tumor syndrome. Patients with this syndrome carry an increased risk of parathyroid and renal tumors. To determine the relevance of HRPT2 for sporadic renal tumors, clear cell, papillary and chromophobe renal cell carcinomas as well as oncocytomas and Wilms tumors were analysed for HRPT2 gene alterations. Loss of heterozygosity (LOH) of HRPT2 was found in seven of 56 (12.5%) clear cell, three of 14 (21%) papillary, six of 10 (60%) chromophobe renal cell carcinomas, three of eight (38%) oncocytomas and four of 10 (40%) Wilms tumors. In addition, two novel HRPT2 point mutations, causing K34Q and R292K changes in parafibromin, were detected in one clear cell carcinoma and one Wilms tumor, respectively. These tumors displayed LOH of the remaining wild-type allele, but interestingly no von Hippel-Lindau (VHL) mutation. Functional analysis revealed that the K34Q mutant species of parafibromin is, unlike wild-type protein, defective in suppressing cyclin D1 expression in vivo. Taken together, these results suggest that renal cancer associated mutations in parafibromin occur in the absence of VHL mutation, which in turn may contribute to constitutively elevated cyclin D1 expression and abnormal cell proliferation. PMID- 17130828 TI - HPV16 E6 confers p53-dependent and p53-independent phenotypes in the epidermis of mice deficient for E6AP. AB - High-risk human papillomaviruses are the causative agents of cervical and other anogenital cancers. In these cancers, two viral oncogenes, E6 and E7, are expressed. E6 is best known for its ability to inactivate the tumor suppressor p53, which is thought to arise through ubiquitin-mediated degradation of p53 and involve a ternary complex between E6, p53 and the E3 ligase, E6AP. In mice transgenic for wild-type HPV16 E6, its expression leads to epithelial hyperplasia and an abrogation of normal cellular responses to DNA damage. Whereas only the latter phenotype is dependent upon E6's inactivation of p53, both are reduced in transgenic mice expressing an E6 mutant severely reduced in its binding to E6AP and other cellular proteins that bind E6 through a shared alpha-helix motif. Here, we investigated whether E6AP is required for the induction of the above phenotypes through the use of both E6AP-mutant and E6AP-null mice. E6, in the absence of E6AP retains an ability to induce epithelial hyperplasia, abrogate DNA damage responses and inhibit the induction of p53 protein following exposure to ionizing radiation. We conclude that E6 is able to induce both p53-dependent and p53-independent phenotypes through E6AP-independent pathways in the mouse. PMID- 17130829 TI - Biochemical characterization of the zinc-finger protein 217 transcriptional repressor complex: identification of a ZNF217 consensus recognition sequence. AB - Zinc-finger protein 217 (ZNF217) is a Kruppel-like zinc-finger protein located at 20q13.2, within a region of recurrent maximal amplification. Here, we demonstrate that ZNF217 is a transcriptional repressor protein and report the purification and characterization of a ZNF217 complex. The purified ZNF217 complex consists of approximately six proteins and contains the transcriptional co-repressors CoREST, BHC110/LSD1, histone deacetylase (HDAC) 2 and C-terminal binding protein (CtBP1). The purified ZNF217 complex possesses deacetylase activity as well as lysine 4 histone H3-specific demethylase activity that is most likely mediated by the BHC110/LSD1 component. To determine if ZNF217 is a sequence-specific binding protein, we have made use of cyclic amplification and selection of targets (CAST) assay and identify for the first time a ZNF217 DNA consensus recognition sequence (CRS) that is highly conserved in the human E-cadherin promoter. Chromatin immunoprecipitation (ChIP) experiments demonstrate that ZNF217, as well as the other components of the ZNF217 complex, are found on the region of the proximal E cadherin promoter that contains the identified ZNF217 CRS in vivo. Using a combination of transient transfections and small interfering RNA, we demonstrate that ZNF217 represses the E-cadherin promoter. Collectively, our results implicate ZNF217 and its associated proteins in a novel pathway that may have profound effects on cancer progression. PMID- 17130830 TI - Combined effects of the two reciprocal t(4;11) fusion proteins MLL.AF4 and AF4.MLL confer resistance to apoptosis, cell cycling capacity and growth transformation. AB - The reciprocal chromosomal translocation t(4;11) is correlated with infant, childhood, adult and therapy-related high-risk acute leukemia. Here, we investigated the biological effects of MLL.AF4, AF4.MLL or the combination of both reciprocal fusion proteins in a conditional in vitro cell culture model system. Several parameters like cell growth, cell cycling capacity, apoptotic behavior and growth transformation were investigated under physiological and stress conditions. Co-transfected cells displayed the highest resistance against apoptotic triggers, cell cycling capacity and loss-of-contact inhibition. These analyses were complemented by gene expression profiling experiments and specific gene signatures were established for each of the three cell lines. Interestingly, co-transfected cells strongly upregulate the homeobox gene Nanog. In combination with Oct4, the Nanog homeoprotein is steering maintenance of pluripotency and self-renewal in embryonic stem cells. Transcription of Nanog and other stem cell factors, like Oct4 and Bmi1, was verified in biopsy material of t(4;11) patient cells which express both reciprocal t(4;11) fusion genes. In conclusion, the presence of both reciprocal MLL fusion proteins confers biological properties known from t(4;11) leukemia, suggesting that each of the two fusion proteins contribute specific properties and, in combination, also synergistic effects to the leukemic phenotype. PMID- 17130831 TI - Cell cycle regulation of the human Six1 homeoprotein is mediated by APC(Cdh1). AB - The Six1 homeoprotein is an important mediator of normal development, where it is critical for the proliferation of precursor cell populations that ultimately constitute the muscle, kidney and inner ear, among other organs. Interestingly, its overexpression has been observed in numerous cancers, where it contributes to the proliferative and metastatic ability of the cancer cells. Here we show that Six1 not only regulates the cell cycle, but is itself regulated throughout the cell cycle via ubiquitin-mediated proteolysis. The protein is present from the G(1)/S boundary until mitosis, when it is degraded via the anaphase-promoting complex (APC) with its activating subunit Cdh1. However, unlike most identified APC(Cdh1) targets, Six1 does not contain functional destruction or KEN box motifs that are necessary for its degradation. Instead, the Six1 protein contains multiple, as yet undefined, sequences within its N- and C-termini responsible for its degradation, including an N-terminal region that binds to Cdh1. Cell cycle regulation of Six1 occurs both transcriptionally and post-translationally via phosphorylation; therefore, this study demonstrates a third and novel mechanism of cell cycle-specific regulation of Six1, underscoring the importance of confining its activity to a defined cell cycle window from the G(1)/S boundary to early mitosis. PMID- 17130832 TI - PDGF essentially links TGF-beta signaling to nuclear beta-catenin accumulation in hepatocellular carcinoma progression. AB - The cooperation of Ras - extracellular signal-regulated kinase/mitogen-activated protein kinase and transforming growth factor (TGF)-beta signaling provokes an epithelial to mesenchymal transition (EMT) of differentiated p19(ARF) null hepatocytes, which is accompanied by a shift in malignancy and gain of metastatic properties. Upon EMT, TGF-beta induces the secretion and autocrine regulation of platelet-derived growth factor (PDGF) by upregulation of PDGF-A and both PDGF receptors. Here, we demonstrate by loss-of-function analyses that PDGF provides adhesive and migratory properties in vitro as well as proliferative stimuli during tumor formation. PDGF signaling resulted in the activation of phosphatidylinositol-3 kinase, and furthermore associated with nuclear beta catenin accumulation upon EMT. Hepatocytes expressing constitutively active beta catenin or its negative regulator Axin were employed to study the impact of nuclear beta-catenin. Unexpectedly, active beta-catenin failed to accelerate proliferation during tumor formation, but in contrast, correlated with growth arrest. Nuclear localization of beta-catenin was accompanied by strong expression of the Cdk inhibitor p16(INK4A) and the concomitant induction of the beta-catenin target genes cyclin D1 and c-myc. In addition, active beta-catenin revealed protection of malignant hepatocytes against anoikis, which provides a prerequisite for the dissemination of carcinoma. From these data, we conclude that TGF-beta acts tumor progressive by induction of PDGF signaling and subsequent activation of beta-catenin, which endows a subpopulation of neoplastic hepatocytes with features of cancer stem cells.. PMID- 17130833 TI - Negative regulation of chemokine receptor CXCR4 by tumor suppressor p53 in breast cancer cells: implications of p53 mutation or isoform expression on breast cancer cell invasion. AB - Chemokine receptor CXCR4 and its ligand CXCL12 are suggested to be involved in migration, invasion and metastasis of breast cancer cells. Mutation of the tumor suppressor gene p53 in breast cancer is associated with metastasis and aggressive clinical phenotype. In this report, we demonstrate that wild type but not the dominant-negative mutant (V143A) or cancer-specific mutants (R175H or R280K) of p53 repress CXCR4 expression. Recently described cancer-specific p53 isoform, Delta133p53, also failed to repress CXCR4 promoter activity. Short-interfering RNA-mediated depletion of p53 increased endogenous CXCR4 expression in MCF-7 breast cancer cells that contain wild-type p53. Basal CXCR4 promoter activity in HCT116 colon carcinoma cells deleted of p53 [HCT116(p53KO)] was 10-fold higher compared to that in parental HCT116 cells with functional wild-type p53. Deletion analysis of CXCR4 promoter identified a seven-base pair p53-repressor element homologous to cyclic AMP/AP-1 response (CRE/AP-1) element. Electrophoretic mobility shift and chromatin immunoprecipitation assays revealed binding of ATF-1 and cJun to the CRE/AP-1 element. The p53 rescue drug PRIMA-1 reduced CXCR4 mRNA and cell surface expression in MDA-MB-231 cells, which express R280K mutant p53. CP-31398, another p53 rescue drug, similarly reduced cell surface levels of CXCR4. PRIMA-1-mediated decrease in CXCR4 expression correlated with reduced invasion of MDA-MB-231 cells through matrigel. These results suggest a mechanism for elevated CXCR4 expression and metastasis of breast cancers with p53 mutations or isoform expression. We propose that p53 rescue drugs either alone or in combination with chemotherapeutic drugs may be effective in reducing CXCR4 mediated metastasis. PMID- 17130834 TI - Rottlerin synergistically enhances imatinib-induced apoptosis of BCR/ABL expressing cells through its mitochondrial uncoupling effect independent of protein kinase C-delta. AB - Although the BCR/ABL tyrosine kinase inhibitor imatinib is highly effective for treatment of chronic myeloid leukemia (CML) and Philadelphia-chromosome positive acute lymphoblastic leukemia (ALL), relapse with emerging imatinib-resistance mutations in the BCR/ABL kinase domain poses a significant problem. Here, we demonstrate that rottlerin, a putative protein kinase C-delta (PKCdelta)-specific inhibitor, acts synergistically with imatinib to induce apoptosis of BCR/ABL expressing K562 and Ton.B210 cells. However, rottlerin inhibited neither PKCdelta nor BCR/ABL in these cells. On the other hand, rottlerin, previously characterized also as a mitochondrial uncoupler, transiently but significantly reduced mitochondrial membrane potential and gradually induced mitochondrial membrane permeabilization. Moreover, two other mitochondrial uncouplers, FCCP and DNP, very similarly induced apoptosis of BCR/ABL-expressing cells in a synergistic manner with imatinib. Imatinib synergistically enhanced mitochondrial membrane permeabilization induced by mitochondrial uncouplers, which led to release of cytochrome c into the cytoplasm and activation of caspases-3 and -9. Rottlerin also enhanced the cytotoxic effect of imatinib in leukemic cells from patients with CML blast crisis and Ph-positive ALL or a cell line expressing the imatinib-resistant E255K BCR/ABL mutant. The present study indicates that rottlerin synergistically enhances imatinib-induced apoptosis through its mitochondrial uncoupling effect independent of PKCdelta and may contribute to the development of new treatment strategy to overcome the imatinib resistance and to cure the BCR/ABL expressing leukemias. PMID- 17130835 TI - PLC gamma contributes to metastasis of in situ-occurring mammary and prostate tumors. AB - Phospholipase C-gamma (PLCgamma) has been implicated in tumor cell motility required for invasiveness and metastasis. Diminished tumor dissemination has been demonstrated in xenograft models, but studies in naturally-occurring tumors are lacking, having been limited by the timing of the interventions. Therefore, we generated mice that express a doxycycline (DOX)-inducible dominant-negative fragment of PLCgamma, PLCz; this approach avoids the in utero lethality caused by the absence of PLCgamma. As we targeted two de novo-occurring carcinomas of the mammary (MMTV-driven polyoma middle T antigen model, PyVmT) and prostate (TRAMP model) glands, we limited expression to these epithelial cells by driving DOX transactivator from the prostatein C3 promoter. This avoids the confounding variable of potentially abrogating motility in stromal and endothelial cells. These mice developed normally in the presence of DOX, except for limited mammary development if treated before 6 weeks and immaturity of the prostate gland if treated before 2 weeks of age. DOX-mediated induction of PLCz from age 8 to 16 weeks in PyVmT mice decreased the number of lung metastases by >10-fold (P<0.06) without a detectable effect on in situ tumor cell proliferation or tumor size. Lung metastases were also significantly decreased in the TRAMP model in which the mice expressed the PLCz fragment (P<0.05). DOX treatment itself had no effect on tumor size or metastasis in control mice, nor did it affect tumor dissemination in nontransgenic littermates. In conclusion, abrogation of the PLCgamma signaling pathway can limit the metastatic potential of carcinomas. PMID- 17130836 TI - Chemokine-mediated migration of melanoma cells towards lymphatics--a mechanism contributing to metastasis. AB - The mechanisms that cause tumors such as melanomas to metastasize into peripheral lymphatic capillaries are poorly defined. Non-mutually-exclusive mechanisms are lymphatic endothelial cell (LEC) chemotaxis and proliferation in response to tumor cells (chemotaxis-lymphangiogenesis hypothesis) or LECs may secrete chemotactic agents that attract cancer cells (chemotactic metastasis hypothesis). Using migration assays, we found evidence supporting both hypotheses. Conditioned medium (CM) from metastatic malignant melanoma (MMM) cell lines attracted LEC migration, consistent with the lymphangiogenesis hypothesis. Conversely, CM from mixed endothelial cells or LECs, but not blood endothelial cells, attracted MMM cells but not non-metastatic melanoma cells, consistent with the chemotactic metastasis hypothesis. MMM cell lines expressed CCR7 receptors for the lymphatic chemokine CCL21 and CCL21 neutralizing antibodies prevented MMM chemotaxis in vitro. To test for chemotactic metastasis in vivo tumor cells were xenotransplanted into nude mice approximately 1 cm from an injected LEC depot. Two different MMM grew directionally towards the LECs, whereas non-metastatic melanomas did not. These observations support the hypothesis that MMM cells grow towards regions of high LEC density owing to chemotactic LEC secretions, including CCL21. This chemotactic metastasis may contribute to the close association between metastasizing tumor cells and peri-tumor lymphatic density and promote lymphatic invasion. PMID- 17130837 TI - Prostaglandin E2-EP4 receptor signalling promotes tumorigenic behaviour of HT-29 human colorectal cancer cells. AB - The predominant product of cyclooxygenase (COX) activity in the colon, prostaglandin (PG) E2 promotes intestinal tumorigenesis. Expression of the PGE2 receptor EP4 is upregulated during colorectal carcinogenesis. Therefore, we investigated the role of elevated PGE2-EP4 receptor signalling in the protumorigenic activity of PGE2 by increasing EP4 receptor expression in HT-29 human colorectal cancer (CRC) cells (HT-29-EP4) by stable transfection. Elevated PGE2-induced EP4 receptor activity in HT-29 cells increased resistance to spontaneous apoptosis and promoted anchorage-independent growth, but had no effect on proliferation of HT-29-EP4 cells. EP4 receptor activation by PGE2 in HT 29-EP4 cells also led to development of fluid-filled cysts, which was associated with increased tight junction protein (occludin and zonula occludens-1) expression. Overexpression of the EP4 receptor in HT-29 cells led to basal EP4 receptor signalling in the absence of exogenous PGE2, which was explained by autocrine activity of endogenous, COX-2-derived PGE2 and constitutive, ligand independent EP4 receptor activity. The predominant signalling pathway mediating antiapoptotic activity downstream of PGE2-EP4 receptor activation in HT-29-EP4 cells was elevation of cyclic adenosine monophosphate (cAMP) levels, which was associated with phosphorylation of cAMP-response element binding protein. EP4 receptor activation led to a small increase in phosphorylated extracellular signal-regulated kinase (ERK) 2 protein levels but inhibition of ERK phosphorylation did not abrogate the antiapoptotic activity of PGE2. However, PGE2-EP4 receptor signalling did not lead to trans-activation of the epidermal growth factor receptor in HT-29 cells. Inhibition of protumorigenic PGE2-EP4 receptor signalling represents a potential strategy for anti-CRC therapy that may avoid the toxicity associated with systemic COX inhibition. PMID- 17130838 TI - The codon 72 polymorphism-specific effects of human p53 are absent in mouse cells: implications on generation of mouse models. AB - Human p53, unlike mouse p53, contains a polymorphic site at codon 72 in exon 4 encoding either an arginine amino acid (72R) or a proline residue (72P). The 72R form was shown to induce apoptosis better than the 72P form, partly owing to its ability to efficiently bind to the nuclear-export protein CRM1 and localize to the mitochondria. This polymorphism has also been associated with cancer predisposition and chemo-sensitivity. Further understanding of the in vivo significance of this polymorphism in carcinogenesis requires the generation of mouse models. We have thus evaluated if the polymorphism-specific effects of human p53 are retained in mouse cells. Though being transcriptionally active, both the human polymorphs were found to have lost their ability to differentially suppress growth and bind to CRM1 or MDM2 in mouse cells. Moreover, chimaeric proteins containing mouse exons 2-3 and human exons 4-11 have also lost the polymorphism-specific effects in human cells, suggesting that human exons 2-3 are important in regulating the polymorphism-specific effects. Furthermore, human p53 and the various chimaeric proteins were generally less effective in inhibiting growth of mouse cells compared to mouse p53, suggesting that mouse p53 is more potent than human p53 in suppressing growth, partly due to enhanced binding of MDM2 to human p53. The data together suggest that mouse cells may not provide an appropriate environment for the manifestation of the polymorphism-specific functional differences of human p53, and hence, cautions against the expression of full-length or chimaeric p53 proteins in mice to study the effects of the polymorphism. PMID- 17130839 TI - Role of 2-5A-dependent RNase-L in senescence and longevity. AB - Senescence is a permanent growth arrest that restricts the lifespan of primary cells in culture, and represents an in vitro model for aging. Senescence functions as a tumor suppressor mechanism that can be induced independent of replicative crisis by diverse stress stimuli. RNase-L mediates antiproliferative activities and functions as a tumor suppressor in prostate cancer, therefore, we examined a role for RNase-L in cellular senescence and aging. Ectopic expression of RNase-L induced a senescent morphology, a decrease in DNA synthesis, an increase in senescence-associated beta-galactosidase activity, and accelerated replicative senescence. In contrast, senescence was retarded in RNase-L-null fibroblasts compared with wild-type fibroblasts. Activation of endogenous RNase-L by 2-5A transfection induced distinct senescent and apoptotic responses in parental and Simian virus 40-transformed WI38 fibroblasts, respectively, demonstrating cell type specific differences in the antiproliferative response to RNase-L activation. Replicative senescence is a model for in vivo aging; therefore, genetic disruption of senescence effectors may impact lifespan. RNase L-/- mice survived 31.7% (P<0.0001) longer than strain-matched RNase-L+/+ mice providing evidence for a physiological role for RNase-L in aging. These findings identify a novel role for RNase-L in senescence that may contribute to its tumor suppressive function and to the enhanced longevity of RNase-L-/- mice. PMID- 17130840 TI - DNA binding and selective gene induction by different forms of the p53 protein. AB - P53 is a tumor suppressor gene that plays a crucial role in suppressing tumorigenesis by inducing either cell cycle arrest or apoptosis in cells with DNA damage. In more than 50% of tumors p53 is inactivated by gene mutations. However, there have also been reports of tumor cells in which p53 remains wild type and is present in elevated concentrations. Here we utilized a set of mutant cell lines, which, unlike the parental A1-5 cell line, which expresses a mouse tsp53 and becomes growth arrested at 32 degrees C, are capable of growth at this same incubation temperature. We found that the tsp53 in the two cell lines, ALTR-17 and ALTR-24, was identical to the parental A1-5s and concentrated in the nucleus at 32 degrees C. Examination of both lines revealed that p21 was induced at 32 degrees C, although to a lesser extent than in parental cells and that the p21 genes were not mutated. Interestingly, evaluation of the conformation of tsp53 using conformation-specific antibodies showed that the protein existed in different forms, which were found to bind DNA using chromatin immunoprecipitation assays and which we showed could induce expression of a p21 reporter construct. We conclude that the tsp53 may exist in various forms capable of binding DNA. PMID- 17130841 TI - Oncogenic K-RAS subverts the antiapoptotic role of N-RAS and alters modulation of the N-RAS:gelsolin complex. AB - Activating mutations in members of the RAS family of genes are among the most common genetic events in human tumorigenesis. Once thought to be functionally interchangeable, it is increasingly recognized that the classical members of this protein family (H-RAS, N-RAS and K-RAS4B) exhibit unique and shared functions that are highly context-dependent. Herein, we demonstrate that the presence of an oncogenic KRAS allele results in elevated levels of GTP-bound N-RAS (N-RAS.GTP) in two human colorectal cancer cell lines, HCT 116 and DLD-1, compared to their isogenic counterparts in which the mutant KRAS allele has been disrupted by homologous recombination. N-RAS subserves an antiapoptotic role in cells expressing wild-type K-RAS; this function is compromised, however, by the presence of mutant K-RAS, and these cells display increased sensitivity to apoptotic stimuli. We additionally identify a physical interaction between N-RAS and gelsolin, a factor that has been shown to promote survival and show that the N-RAS:gelsolin complex is modulated differently in wild-type and mutant K-RAS environments following apoptotic challenge. These findings represent the first biochemical evidence of a functional relationship between endogenous RAS proteins and identify a dynamic physical interaction between endogenous N-RAS and gelsolin that correlates with survival. PMID- 17130842 TI - eEF1A2 activates Akt and stimulates Akt-dependent actin remodeling, invasion and migration. AB - eEF1A2 (eukaryotic protein elongation factor 1 alpha 2) is a protein translation factor that is likely a human oncogene by virtue of its capacity to transform mammalian cells and its high expression in tumors of the ovary, breast and lung. Here, we show that expression of eEF1A2 is sufficient to stimulate the formation of filopodia in BT549 human breast cancer cells and non-transformed Rat2 cells. Filopodia formation in eEF1A2-expressing cells is dependent on the activity of phosphatidylinositol-3 kinase (PI3K), and the ROCK and Akt kinases. Furthermore, eEF1A2 expression is sufficient to activate Akt in a PI3K-dependent fashion and inactivation of eEF1A2 by short interfering RNA reduces Akt activity. Using breast cancer cell line BT 549, we show that eEF1A2 expression stimulates cell migration and invasion in a largely PI3K- and Akt-dependent manner. These results suggest that eEF1A2 regulates oncogenesis through Akt and PI3K-dependent cytoskeletal remodeling. PMID- 17130843 TI - Lysophosphatidic acid downregulates tissue inhibitor of metalloproteinases, which are negatively involved in lysophosphatidic acid-induced cell invasion. AB - Ovarian cancer is a highly metastatic disease. Lysophosphatidic acid (LPA) levels are elevated in ascites from ovarian cancer patients, but its potential role in ovarian cancer metastasis has just begun to be revealed. In this work, we show that LPA stimulates invasion of primary ovarian cancer cells, but not ovarian epithelial or borderline ovarian tumor cells, although these benign cells indeed respond to LPA in cell migration. We have found that LPA downregulates tissue inhibitor of metalloproteinases (TIMPs). TIMP2 and TIMP3 play functional role in LPA-induced invasion as negative regulators. G(i) protein, phosphatidylinositol-3 kinase (PI3K), p38 mitogen-activated protein kinase (MAPK), cytosolic phospholipase A(2) and urokinase type plasminogen activator (uPA) are required for LPA-induced cells invasion. TIMP3 may affect two independent downstream targets, vascular endothelial growth factor receptor and p38 MAPK. In vivo, LPA stimulates tumor metastasis in an orthotopic ovarian tumor model, which can be inhibited by a PI3K inhibitor, LY294002. In summary, LPA is likely a key component for promoting ovarian metastasis in vivo. LPA downregulates TIMP3, which may have targets other than metalloproteinases. Our in vivo metastasis mouse model is useful for studying the efficacy of therapeutic regimes of ovarian cancer. PMID- 17130844 TI - Cross-talk between calpain and caspase-3/-7 in cisplatin-induced apoptosis of melanoma cells: a major role of calpain inhibition in cell death protection and p53 status. AB - The contribution of different proteolytic systems, in particular calpains and effector caspases, in apoptotic cell death is still controversial. In this paper, we show that during cisplatin-induced apoptosis of human metastatic melanoma cells, calpain activation, as measured in intact cells by two different fluorescent substrates, is an early event, taking place well before caspase-3/-7 activation, and progressively increasing during 48 h of treatment. Such activation appears to be independent from any intracellular calcium imbalance; in fact, an increase of cytosolic calcium along with emptying of the reticular stores occur only at very late stages, uniquely in frankly apoptotic, detached cells. Calpain activation proves to be an early and crucial event in the apoptotic machinery, as demonstrated by the significant protection of cell death in samples co-treated with the calpain inhibitors, MDL 28170, calpeptin and PD 150606, where a variable but significant reduction of both caspase-3/-7 activity and cell detachment is observed. Consistently, such a protective effect can be at least partially due to the impairment of cisplatin-induced p53 activation, occurring early in committed, preapoptotic cells. Furthermore, in late apoptotic cells, calpain activity is also responsible for the formation of a novel p53 proteolytic fragment (approximately 26 kDa), whose function is so far to be elucidated. PMID- 17130845 TI - Serine protease Omi/HtrA2 targets WARTS kinase to control cell proliferation. AB - The serine protease Omi/HtrA2 was initially regarded as a proapoptotic molecule that proteolyses several proteins to induce cell death. Recent studies, however, indicate that loss of Omi protease activity increases susceptibility to stress induced cell death. These complicated findings suggest that the protease activity of Omi is involved not only in apoptosis but also in cellular homeostasis. However, the targets which Omi uses to mediate this novel process are unknown. Previously, we showed that WARTS (WTS)/large tumor-suppressor 1 mitotic kinase interacts with the protein/discs-large protein/zonula (PDZ) domain of Omi and promotes its protease activity. We now report that WTS is a substrate for Omi protease activity, thus it is not only a regulator but also a downstream target of this protease. Interaction with Omi PDZ domain is required for WTS to be proteolysed. When caspase-9-deficient mouse embryonic fibroblasts (MEFs) were treated with staurosporine, WTS was proteolysed by activated endogenous Omi without induction of cell death. Therefore, protease activity of Omi and proteolysis of WTS are not necessarily required for cell death. We found that depletion of Omi from HeLa cells results in accelerated cell proliferation despite no significant change in the duration of mitosis. The depletion of WTS showed the same effect on S phase progression. Therefore, WTS proteolytic fragment(s) generated by Omi may act as an inhibitor of G1/S progression. Our data reveal a role for Omi-mediated processing of WTS in negative regulation of cell cycle progression at interphase, suggesting a novel function of Omi other than apoptosis. PMID- 17130846 TI - Evolution of plant microRNA gene families. AB - MicroRNAs (miRNAs) are important post-transcriptional regulators of their target genes in plants and animals. miRNAs are usually 20-24 nucleotides long. Despite their unusually small sizes, the evolutionary history of miRNA gene families seems to be similar to their protein-coding counterparts. In contrast to the small but abundant miRNA families in the animal genomes, plants have fewer but larger miRNA gene families. Members of plant miRNA gene families are often highly similar, suggesting recent expansion via tandem gene duplication and segmental duplication events. Although many miRNA genes are conserved across plant species, the same gene family varies significantly in size and genomic organization in different species, which may cause dosage effects and spatial and temporal differences in target gene regulations. In this review, we summarize the current progress in understanding the evolution of plant miRNA gene families. PMID- 17130847 TI - Prevention of high-fat diet-induced adipose tissue remodeling in obese diabetic mice by n-3 polyunsaturated fatty acids. AB - OBJECTIVE: Obesity is associated with reduced insulin sensitivity and extensive reorganization of adipose tissue. As polyunsaturated fatty acids (PUFA) appear to inhibit diabetes development, we investigated PUFA effects on markers of matrix remodeling in white adipose tissue. METHODS AND PROCEDURE: Male obese diabetic (db/db) mice were treated with either a low-fat standard diet (LF), or high-fat diets rich in saturated and monounsaturated fatty acids (HF/S), n-6 PUFA (HF/6) or the latter including marine n-3 PUFA (HF/3). White adipose tissue was analyzed for gene expression, fatty acid composition and by immunofluorescence. RESULTS: HF/S treatment increased adipose tissue expression of a number of genes involved in matrix degradation including matrix metalloproteinase (MMP)-12, -14 and cathepsin K, L and S compared with LF. MMP-12 gene was expressed in macrophages and adipocytes, and MMP-12 protein colocalized with both cell types. In addition, mean adipocyte area increased by 1.6-fold in HF/S-treated mice. Genes essential for collagen production, such as procollagen I, III, VI, tenascin C and biglycan were upregulated in HF/S-treated animals as well. N-3 PUFA supplementation resulted in enrichment of these fatty acids in adipose tissue. Moreover, n-3 PUFA inhibited the HF/S-induced upregulation of genes involved in matrix degradation and production I restored mean adipocyte area and prevented MMP-12 expression in macrophages and adipocytes. CONCLUSION: N-3 PUFA prevent high-fat diet-induced matrix remodeling and adipocyte enlargement in adipose tissue of obese diabetic mice. Such changes could contribute to diabetes prevention by n-3 PUFA in obese patients. PMID- 17130848 TI - Affordable measurement of human total energy expenditure and body composition using one-tenth dose doubly labelled water. AB - BACKGROUND: The doubly labelled water (DLW) method is the technique of choice for measurement of free-living total energy expenditure (TEE) in humans. A major constraint on the clinical applicability of the method has been the expense of the (18)O isotope. METHOD: We have used a reduced-dose (one-tenth of the currently recommended standard dose) of DLW for the measurement of TEE and body composition in nine healthy adult male volunteers. RESULTS: TEE measured by reduced-dose DLW was positively correlated with resting energy expenditure measured by metabolic cart (r=0.87, P<0.01). Isotope-derived fat mass and body mass index were strongly correlated (r=0.86, P<0.01). In four subjects in whom we performed a complementary evaluation using standard-dose isotope enrichment, the TEE measurements were satisfactorily comparable (mean+/-s.d.: reduced dose 2586+/ 155 kcal/day vs standard dose 2843+/-321 kcal/day; mean difference 257+/-265 kcal/day). CONCLUSION: These data indicate that DLW measurements of human energy expenditure and body composition can be performed at a substantially reduced dose (and cost) of isotope enrichment than is currently employed. PMID- 17130849 TI - Using intranasal lidocaine to reduce food intake. AB - OBJECTIVE: Develop a dose-response curve for the effect of intranasal lidocaine on food intake. DESIGN: Healthy obese subjects had food intake, ratings of hunger, desire to eat, craving and fullness measured at lunch after an overnight fast. Four treatments were given as nose drops (0.5-0.6 ml per nostril) 5 min before the meal in a double-blind manner with a four period crossover design including a 7-day washout between periods. The treatments were saline, 2.5, 10 and 25 mg lidocaine per nostril. The order of administration was randomly assigned to each subject. Electrocardiograms, vital signs, chemistry panels, complete blood counts (CBC) and nasal inspections were carried out before and after each dose. SUBJECTS: Forty-seven subjects were screened, 34 were randomized and 20 subjects completed all four study periods in the trial. The subjects were 39+/-12.5 (s.d) years of age, had a weight of 91+/-13.0 kg, a height of 167+/ 10.3 cm, 56% were women, 47% were African-American and 53% were Caucasian. MEASUREMENTS: Food intake, rating of hunger, desire to eat, craving and fullness are measures of efficacy. Adverse events, electrocardiograms, vital signs, chemistry panels, nasal inspections, CBC and physical exams are measures of safety. RESULTS: The mean reduction in food intake vs saline control in the 20 subjects completing all four study periods was 3.3+/-7% (s.d), 4.2+/-8.5% and 7.4+/-7.3% in the 2.5 mg, 10 and 25 mg per nostril groups, respectively (P=NS). Hunger and desire to eat in subjects who completed at least one study period decreased dose dependently (P<0.03, at the 25 mg per nostril dose). There were no clinically significant changes in safety measures, electrocardiograms, vital signs, chemistry panels, CBC or nasal inspections. CONCLUSION: Intranasal lidocaine reduced hunger and the desire to eat, but this did not translate into a significant reduction in food intake suggesting that intranasal lidocaine will not have value in treating obesity. PMID- 17130850 TI - Disordered eating behaviours and cognitions in young women with obesity: relationship with psychological status. AB - OBJECTIVE: To examine levels of eating disorder behaviours and cognitions of young women with obesity in the Australian Capital Territory, Australia and assess the impact upon psychological status. DESIGN: General population cross sectional survey. SUBJECTS: A total of 4891 young women from the community aged 18-42 years, of which 630 were in the obese weight range. MEASUREMENTS: Body mass index (BMI), eating disorder psychopathology (eating disorder examination questionnaire), and psychological distress (K-10). RESULTS: Women with obesity had significantly higher levels of dietary restraint, eating concern, weight concern, shape concern, binge eating, misuse of diuretics, use of diet pills and fasting compared to other women in the community. These eating disorder cognitions and behaviours were associated with increased levels of psychological distress. In women with obesity, eating concern, weight concern, shape concern, dietary restraint and decreased age predicted psychological distress in a multivariate model. Among other women in the community, behaviours such as laxative misuse, 'hard' exercise and subjective bulimic episodes also contributed to the model predicting psychological distress. CONCLUSION: As disordered eating psychopathology is high in young obese women and negatively impacts upon psychological status, obesity prevention and treatment should consider eating disorder psychopathology and mental health outcomes. PMID- 17130851 TI - The entero-insular axis and adipose tissue-related factors in the prediction of weight gain in humans. AB - Obesity has now reached epidemic proportions. Epidemiological studies in the past decades have shown that adults gain weight and adiposity from the early twenties until their sixties. In the paediatric population, growing numbers of children and adolescents put on unhealthy weight. Many environmental, socio-economical and biological determinants that predispose to weight gain have been identified thus far. The aim of the present review is to summarize the current knowledge on the role of the circulating levels of adipokines and other entero-insular hormones and biological markers of obesity to predict weight gain in humans. The review focuses on relationship between hormonal and biochemical markers (insulin, insulin-like growth factors, gastrointestinal hormones, leptin, adiponectin, resistin, inflammatory proteins and cytokines) and weight gain in prospective studies. The complex relationships displayed by these hormonal factors with future weight gain in humans are critically reviewed and integrative models are proposed. Overall, most of the studies reported to date made adjustments for baseline body mass index but failed to consider dietary intake and physical activity as confounding factors. Outstanding questions are raised and new directions for future prospective studies are proposed in order to improve our understanding of the role of biological determinants of energy balance and development of obesity in humans. PMID- 17130852 TI - Association of a beta-2 adrenoceptor (ADRB2) gene variant with a blunted in vivo lipolysis and fat oxidation. AB - BACKGROUND AND AIMS: Obesity is associated with a blunted beta-adrenoceptor mediated lipolysis and fat oxidation. We investigated whether polymorphisms in codon 16, 27 and 164 of the beta (2)-adrenoceptor gene (ADRB2) and exon 10 of the G protein beta (3)-subunit gene (GNB3) are associated with alterations in in vivo lipolysis and fat oxidation. DESIGN AND METHODS: Sixty-five male and 43 female overweight and obese subjects (body mass index (BMI) range: 26.1-48.4 kg/m(2)) were included. Energy expenditure (EE), respiratory quotient (RQ), circulating free fatty acid (FFA) and glycerol levels were determined after stepwise infusion of increasing doses of the non-selective beta-agonist isoprenaline (ISO). RESULTS: In women, the Arg16 allele of the ADRB2 gene was associated with a blunted increase in circulating FFA, glycerol and a decreased fat oxidation during ISO stimulation. In men, the Arg16 allele was significantly associated with a blunted increase in FFA but not in glycerol or fat oxidation. CONCLUSION: These results suggest that genetic variation in the ADRB2 gene is associated with disturbances in in vivo beta-adrenoceptor-mediated lipolysis and fat oxidation during beta-adrenergic stimulation in overweight and obese subjects; these effects are influenced by gene-gender interactions. PMID- 17130866 TI - Dynamics of single-cell gene expression. AB - Cellular behavior has traditionally been investigated by utilizing bulk-scale methods that measure average values for a population of cells. Such population wide studies mask the behavior of individual cells and are often insufficient for characterizing biological processes in which cellular heterogeneity plays a key role. A unifying theme of many recent studies has been a focus on the development and utilization of single-cell experimental techniques that are capable of probing key biological phenomena in individual living cells. Recently, novel information about gene expression dynamics has been obtained from single-cell experiments that draw upon the unique capabilities of fluorescent reporter proteins. PMID- 17130867 TI - Communication between levels of transcriptional control improves robustness and adaptivity. AB - Regulation of eukaryotic gene expression depends on groups of related proteins acting at the levels of chromatin organization, transcriptional initiation, RNA processing, and nuclear transport. However, a unified understanding of how these different levels of transcriptional control interact has been lacking. Here, we combine genome-wide protein-DNA binding data from multiple sources to infer the connections between functional groups of regulators in Saccharomyces cerevisiae. Our resulting transcriptional network uncovers novel biological relationships; supporting experiments confirm new associations between actively transcribed genes and Sir2 and Esc1, two proteins normally linked to silencing chromatin. Analysis of the regulatory network also reveals an elegant architecture for transcriptional control. Using communication theory, we show that most protein regulators prefer to form modules within their functional class, whereas essential proteins maintain the sparse connections between different classes. Moreover, we provide evidence that communication between different regulatory groups improves the robustness and adaptivity of the cell. PMID- 17130868 TI - Biological context networks: a mosaic view of the interactome. AB - Network models are a fundamental tool for the visualization and analysis of molecular interactions occurring in biological systems. While broadly illuminating the molecular machinery of the cell, graphical representations of protein interaction networks mask complex patterns of interaction that depend on temporal, spatial, or condition-specific contexts. In this paper, we introduce a novel graph construct called a biological context network that explicitly captures these changing patterns of interaction from one biological context to another. We consider known gene ontology biological process and cellular component annotations as a proxy for context, and show that aggregating small process-specific protein interaction sub-networks leads to the emergence of observed scale-free properties. The biological context model also provides the basis for characterizing proteins in terms of several context-specific measures, including 'interactive promiscuity,' which identifies proteins whose interacting partners vary from one context to another. We show that such context-sensitive measures are significantly better predictors of knockout lethality than node degree, reaching better than 70% accuracy among the top scoring proteins. PMID- 17130869 TI - A generic problem. PMID- 17130870 TI - What are the options for upgrading the quality of clinical assessment and research in Barrett's esophagus? PMID- 17130871 TI - Do PPIs diminish Barrett's esophagus length or cancer risk? PMID- 17130872 TI - What are the risk factors for surgery and their implications for management of children with Crohn's disease? PMID- 17130873 TI - Minilaparotomy cholecystectomy or laparoscopic cholecystectomy for symptomatic gallstones? PMID- 17130874 TI - Does mycophenolate mofetil reduce the risk of late acute rejection after liver transplantation? PMID- 17130875 TI - How useful is EUS in patients with gastric cancer? PMID- 17130876 TI - Laparoscopic or open appendectomy for complicated appendicitis? PMID- 17130877 TI - Genetic testing for colon cancer. AB - Colon cancer remains the third leading cause of death due to cancer in the US, where it affected more than 145,000 individuals in 2005. Up to 30% of these cases exhibit familial clustering, which means that tens of thousands of individuals have a disease with a potentially definable genetic component. Approximately 3-5% of colon cancers are associated with high-risk, inherited colon cancer syndromes. Identification of the genes that cause these colon cancer syndromes, coupled with additional insights into their clinical course, has led to the development of specific management guidelines--and genetic tests--that can diagnose these familial disorders. These guidelines can be life-saving, not only for the affected patient, but also for their family members. PMID- 17130878 TI - Post-ERCP pancreatitis and its prevention. AB - Pancreatitis remains the most common severe complication of endoscopic retrograde cholangiopancreatography (ERCP), and typically develops in 5-7% of patients. Although most post-ERCP pancreatitis (PEP) is mild, severe pancreatitis and its complications (including a systemic inflammatory response or the development of pseudocysts or pancreatic necrosis) can occur, and in rare cases death can result. A means of preventing PEP in all patients who undergo the procedure remains elusive. Proper patient selection for ERCP is critical to avoid unnecessary risk. Pharmacologic attempts to prevent PEP have been largely unsuccessful; encouraging results have been difficult to validate. Prophylactic stenting of the pancreatic duct and minimally traumatic cannulation techniques offer the most promise as a means of preventing PEP. This manuscript reviews risk factors for PEP as well as pharmacologic and procedural means that can be used to reduce its incidence. PMID- 17130879 TI - Vitamin D replacement for cirrhosis-related bone disease. AB - The osteoporotic fracture rate in patients with chronic liver disease is approximately twice that of age-matched, control individuals. About 66% of patients with moderately severe cirrhosis and 96% of patients awaiting liver transplantation have vitamin D deficiency. Studies have shown a strong correlation between vitamin D deficiency and bone density, particularly in the hip. Previous studies of vitamin D therapy in cirrhosis-related bone disease have had major design flaws. Most reports and guidelines on the treatment of hepatic bone disease have concluded that vitamin D deficiency does not have a significant pathogenetic role in the development of osteoporosis in cirrhosis, and that there is no evidence for a therapeutic effect of vitamin D supplementation. Conversely, it is generally recommended that patients with cirrhosis and low bone density should receive calcium and vitamin D supplementation; yet there is a paucity of reliable data on the optimal doses to use, as well as a lack of clearly demonstrated benefit. We believe that clinical trials of vitamin D therapy in these patients with liver disease are warranted. As low-dose oral supplementation often will not normalize vitamin D levels or suppress parathyroid hormone activity in cirrhotic patients, high-dose, parenteral vitamin D might be preferable, but further long-term studies are required to assess the benefits and safety of this approach. PMID- 17130880 TI - Schistosomal colitis without granuloma formation in a kidney transplant recipient. AB - BACKGROUND: A 40-year-old male from the White Nile region in Sudan, who had received a kidney transplant 6 years previously, presented with fever, lower abdominal pain and diarrhea stained with blood of 5 months duration. He was on immunosuppressive maintenance therapy, consisting of ciclosporin 75 mg twice daily, prednisolone 10 mg once daily, and azathioprine 75 mg once daily. INVESTIGATIONS: Laboratory investigations, liver function tests, renal function tests, stool microscopy, stool culture, abdominal ultrasound, and colonoscopy. DIAGNOSIS: Severe, left-sided colitis due to Schistosoma mansoni infection, without granuloma formation. MANAGEMENT: Oral antischistosomal therapy with praziquantel at a dose of 40 mg/kg body weight. PMID- 17130882 TI - Further evidence for association between ErbB4 and schizophrenia and influence on cognitive intermediate phenotypes in healthy controls. PMID- 17130883 TI - No association between dopaminergic polymorphisms and intelligence variability in attention-deficit/hyperactivity disorder. PMID- 17130884 TI - Expression of ribosomal subunit genes increased coordinately with postmortem interval in human brain. PMID- 17130885 TI - 150 years of Sigmund Freud: What would Freud have said about the obesity epidemic? AB - The 150th birthday of Sigmund Freud has triggered widespread interest and media coverage on his unique contribution and impact on society. Recent evidence from neuroscience and advanced imaging technology has provided support for some of his major concepts including the unconscious and the key role of early life events. In this perspective, we attempt to write on his behalf an updated version of a Freudian way of thinking focused on the current high rates of obesity and depression. PMID- 17130888 TI - Cervical spinal cord neurapraxia in the setting of Klippel-Feil anomaly: a diagnostic and therapeutic challenge. AB - STUDY DESIGN: Case report. SETTING: Temple University Children's Medical Center in USA. OBJECTIVES: To report a patient whose recurrent and transient episodes of quadriplegia mimicked cervical cord neurapraxia. Cervical spine neuroimaging revealed congenital intersegmental fusion of C5 through C7 (consistent with Klippel-Feil anomaly), corresponding abnormal spinal cord signals on T2-weighted images and enhancing focal lesion opposite the C4 vertebral body. A posterior cervical decompression at C4-C5 and lateral mass fixation was performed, and the episodic quadriplegia has not recurred. CONCLUSION: Understanding of the biomechanics of Klippel-Feil anomaly may facilitate prompt referral for surgical management and avoidance of permanent disability. PMID- 17130889 TI - Maximal static mouth respiratory pressure in spinal cord injured patients: correlation with motor level. AB - STUDY DESIGN: Transversal. OBJECTIVES: The few studies concerning maximal static mouth respiratory pressures in patients with spinal cord lesions suggest a marked reduction. We studied the correlation of these parameters with the motor level of injury. SETTING: Rehabilitation Center, Brasilia/DF, Brazil. METHODS: One hundred and thirty-one patients with traumatic spinal cord injury (C4-L3) were recruited. The participants were assessed by standard spirometry and maximal static mouth respiratory pressure. RESULTS: Forced vital capacity was most reduced in tetraplegics (subgroup C4-C5, 49%+/-25 predicted) and increase successively for each descending subgroup (C6-C8, 61%+/-22 predicted; T1-T6, 70%+/-15 predicted), becoming normal in low paraplegia (T7-L3, 84%+/-15 predicted). There is no evidence of an obstructive disturbance throughout all groups. The lowest average percent predicted of maximal static inspiratory pressure (MIP) was in the subgroup C4-C5 (50%+/-23). The average percent predicted of maximal static expiratory pressure (MEP) improved from 19%+/-14 in the C4-C5 subgroup to 51%+/ 19 for T7-L3 subgroup. The average percent predicted of all participants for MIP was 74%+/-30 and for MEP was 37%+/-21. In patients with complete motor lesion, the correlation with the level of injury was stronger for MEP (r=0.81, P<0.0001; r (2)=0.65) than for MIP (r=0.62, P=0.004; r (2)=0.38). No correlation was found among incomplete motor lesion patients. CONCLUSIONS: The linear regression equations for the relationship of percent predicted MIP or MEP to level of injury are applicable only to complete motor lesions and may be useful to establish normative association between them. PMID- 17130891 TI - Blunt cervical spine trauma as a cause of spinal cord injury and delayed cortical blindness. AB - STUDY DESIGN: Case report. OBJECTIVE: To present and discuss the case of a patient who sustained a significant flexion compression injury of the cervical spine with resulting tetraplegia and development of cortical blindness. SETTING: National Spinal Injuries Unit and Institute of Neurological Sciences, Southern General Hospital, Glasgow, Scotland, UK. METHODS: Clinical and radiological follow-up of the patient. RESULTS: Cortical blindness resulted from vertebral artery dissection associated with blunt cervical spine trauma. The patient is registered blind and is ventilator dependent. CONCLUSION: The potential complications of blunt vertebral artery injury remain poorly recognised. Screening is routinely not performed. Advances in noninvasive radiological techniques may result in recognition of asymptomatic disease and the potential for therapeutic intervention. PMID- 17130892 TI - Thermoregulation in tetraplegic patients. PMID- 17130896 TI - Autoimmunity: red line to arthritis. PMID- 17130897 TI - Host and viral factors in the immunopathogenesis of primary hepatitis C virus infection. AB - Individuals infected with hepatitis C virus (HCV) have two possible outcomes of infection, clearance or persistent infection. The focus of this review is the host mechanisms that facilitate clearance. The interaction between HCV viral components and the immune system ultimately determines the balance between the virus and host. Strong evidence points to the aspects of cellular immune response as the key determinants of outcome. The recent discovery of viral evasion strategies targeting innate immunity suggests that the interferon-alpha/beta induction pathways are also critical. A growing body of evidence has implicated polymorphisms in both innate and adaptive immune response genes as determinants of viral clearance in individuals infected with HCV. PMID- 17130898 TI - Immunopathogenesis of hepatitis B virus infection. AB - Hepatitis B virus (HBV) infection is a non-cytopathic hepatotropic virus that can lead to severe liver disease including acute hepatitis, cirrhosis and hepatocellular carcinoma. Successful clearance of the virus as well as the establishment of liver disease is largely driven by a complex interaction between the virus and the host immune response. In this review, the immunological events, including both the innate and adaptive immune response are discussed in the setting of both acute and chronic HBV infection and liver disease. PMID- 17130899 TI - Of cascades and perfect storms: the immunopathogenesis of dengue haemorrhagic fever-dengue shock syndrome (DHF/DSS). AB - The past four decades has witnessed a consolidation of the original observations made in the 1970s that dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS) have an immunological basis. Following reinfection with a dengue virus of different serotype, severe disease is linked to high levels of antibody-enhanced viral replication early in illness which is followed by a cascade of memory T cell activation and a 'storm' of inflammatory cytokines and other chemical mediators. These compounds are released mainly from T cells, monocytes/macrophages and endothelial cells, and ultimately cause an increase in vascular permeability. The consolidation of the evidence has been largely due to several important prospective sero-epidemiological studies in areas endemic for DHF/DSS, which have shown that risk of severe disease is significantly higher in secondary dengue infections. These advances have underscored the fact that DHF/DSS pathogenesis is a complex, multifactorial process involving cocirculation of various dengue virus serotypes and the interplay of host and viral factors that influence disease severity. The continued search to define risk factors in susceptible populations must be combined with the new techniques of molecular virology and innovative approaches in vaccine design to achieve the ultimate objective of developing a safe and effective vaccine. PMID- 17130900 TI - Immunoepigenetics: the unseen side of cancer immunoediting. AB - Cancer immunosurveillance representing, till recently, the explanatory framework relating cancer and the immune system, does not convincingly explain tumor escape. At the beginning of the decade, a new theory emerged, namely the immunoediting theory, and it comprehensively defines the role of the immune system in carcinogenesis. The core of this theory embraces the concept that the immune system on the one hand protects the body from cancer and on the other it shapes the immunogenicity of these cancers, thus presents a persuasive rationalization of the resistance of tumors against the immune response. With the immune system playing, in this context, such a pivotal role in shaping the tumor immune profile and in subsequent oncogenesis, it seems rather paradoxical to accept the immunocompetent host's immune system as a constant moiety. While DNA mutations of immune genes create a rather polymorphic condition, their frequency is much lower than that of other genetic events. Of these, epigenetic alterations give rise to new epialleles, which can reach up to 100% per locus. Bearing in mind that cancer is characterized by a tremendous amount of epigenetic aberrations, in both gene and global level, it is reasonable to postulate that, for the same unknown causes, analogous aberrations could affect the immune genes. Should this be the case, the relation between oncogenesis and the immune system appears much more dynamic and complex. Such an immunoepigenetic approach to carcinogenesis could improve our understanding of a series of common cancer related aspects, such as environmental risk factors, effectiveness of demethylating agents, failure of current immunotherapies, etc. Moreover, this immunoepigenetic paradigm will take the current perception of the immune system and cancer interrelation further and beyond, constituting that the immunoresistant cancer cell phenotype is not shaped by the immune system acting as a steady and rigid evolutionary pressure, but rather as an extremely dynamic variable. PMID- 17130901 TI - Rottlerin inhibits P2X(7) receptor-stimulated phospholipase D activity in chronic lymphocytic leukaemia B-lymphocytes. AB - Phospholipase D (PLD) is a ubiquitous enzyme that can be activated by extracellular adenosine 5'-triphosphate (ATP) or phorbol 12-myristate 13-acetate (PMA) in B-lymphocytes from subjects with chronic lymphocytic leukaemia (CLL). In this study, ATP- but not PMA-induced PLD stimulation in CLL B-lymphocytes was abolished in the presence of an anti-P2X(7) receptor monoclonal antibody, as well as in B-lymphocytes from CLL subjects homozygous for the Glu(496) to Ala loss-of function P2X(7) polymorphism. Rottlerin, an inhibitor of novel protein kinase C (PKC) isoforms, but not GF 109203X, an inhibitor of conventional PKC isoforms, impaired the ATP-stimulated PLD activity in CLL B-lymphocytes. In contrast, both inhibitors impaired PLD activity stimulated by PMA, a known mediator of PKC activation. The inhibition of P2X(7)-stimulated PLD activity by rottlerin was attributed to a target downstream of P2X(7) activation, as the ATP-mediated (86)Rb(+) efflux from CLL B-lymphocytes was not altered in the presence of rottlerin. Our results indicate a possible role for novel PKC isoforms in the regulation of P2X(7)-mediated PLD activity. PMID- 17130902 TI - Positive reactions on Western blots do not necessarily indicate the epitopes on antigens are continuous. AB - Epitope mapping (identification of an antigenic site recognized by an antibody) is an important component of vaccine development and immunological assays. It is widely accepted that in Western blots, antibodies react exclusively with continuous epitopes: discontinuous epitopes are assumed to be irreversibly destroyed by electrophoresis under the denaturing conditions used for sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Here, we demonstrate that the epitopes recognized by four different monoclonal antibodies were identified as discontinuous epitopes when characterized by radioimmunoprecipitation assays and enzyme-linked immunosorbent assays, yet each of these antibodies reacted with the corresponding antigen on Western blots. Reaction on Western blots may be due to epitope renaturation during or after the transfer of the protein to a membrane. Therefore, positive reactions on Western blots do not necessarily indicate that epitopes are continuous and this caveat should be kept in mind while characterizing them. PMID- 17130903 TI - Upregulation of MIP-2 (CXCL2) expression by 15-deoxy-Delta(12,14)-prostaglandin J(2) in mouse peritoneal macrophages. AB - A peroxisome proliferator-activated receptor gamma (PPARgamma) ligand, 15-deoxy Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), has been reported to possess anti inflammatory activity in activated monocytes/macrophages. In this study, we investigated the effect of 15d-PGJ(2) on the lipopolysaccharide (LPS)-induced expression of chemokine mRNAs, especially macrophage inhibitory protein (MIP)-2 (CXCL2), in mouse peritoneal macrophages. The inhibitory actions of the natural PPARgamma ligands, 15d-PGJ(2) and prostaglandin A1 (PGA1), on the expression of RANTES (regulated upon activation, normal T expressed and secreted; CCL5), MIP 1beta (CCL4), MIP-1alpha (CCL3), IFN-gamma-inducible protein 10 kilodaltons (IP 10; CXCL10) and monocyte chemoattractant protein-1 (MCP-1; CCL2) mRNA in LPS treated cells were stronger than those of the synthetic PPARgamma ligands troglitazone and ciglitazone. However, 15d-PGJ(2) enhanced the expression of LPS induced MIP-2 (CXCL2) mRNA. A specific PPARgamma antagonist (GW9662) had no effect on the inhibitory action of 15d-PGJ(2) and PGA1 in LPS-induced chemokine mRNA expression and on the synergistic action of 15d-PGJ(2) in LPS-induced MIP-2 (CXCL2) expression. Moreover, LPS itself reduced the expression of PPARgamma. Although the synergistic effect of 15d-PGJ(2) on LPS-induced MIP-2 (CXCL2) mRNA expression was remarkable, the production of MIP-2 (CXCL2) in cells treated with 15d-PGJ(2) and LPS did not increase compared to the production in cells treated with LPS alone. The synergistic action of 15d-PGJ(2) on LPS-induced MIP-2 (CXCL2) mRNA expression was dependent on the activation of nuclear factor-kappaB (NF kappaB), and 15d-PGJ(2) increased the phosphorylation of p38 and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) in cells stimulated with LPS. These results suggest that the synergistic effect of 15d-PGJ(2) on LPS-induced MIP-2 (CXCL2) expression is PPARgamma-independent, and is mediated by the p38 and SAPK/JNK pathway in mitogen-activated protein kinase signaling pathways, which activates NF-kappaB. Our data may give more insights into the different mechanisms contrary to the anti-inflammatory effect of 15d-PGJ(2) on the expression of chemokine genes. PMID- 17130904 TI - Cramer-Rao lower bound for the estimation of the degree of polarization in active coherent imagery at low photon levels. AB - The degree of polarization (DOP) is an important tool in many optical measurement and imaging applications. We address the problem of its estimation in images that are perturbed with both speckle and photon noise, by determining the Cramer-Rao lower bounds (CRLBs) when the illuminated materials are purely depolarizing. We demonstrate that the CRLBs are simply the sum of the CRLBs due to speckle noise and Poisson noise. We use this result to analyze the influence of different optical parameters on DOP estimation. PMID- 17130905 TI - Active Q switching of a fiber laser with a microsphere resonator. AB - We propose and demonstrate an active Q-switched fiber laser using a high-Q microsphere resonator as the Q-switching element. The laser cavity consists of an Er-doped fiber as the gain medium, a glass microsphere reflector (coupled through a fiber taper) at one end of the cavity, and a fiber Bragg grating reflector at the other end. The reflectivity of the microsphere is modulated by changing the gap between the microsphere and the fiber taper. Active Q switching is realized by oscillating the microsphere in and out of contact with the taper. Using this novel technique, we have obtained giant pulses (maximum peak power approximately 102 W, duration approximately 160 ns) at a low pump-power threshold (approximately 3 mW). PMID- 17130906 TI - Wavelength-spacing-tunable multichannel filter incorporating a sampled chirped fiber Bragg grating based on a symmetrical chirp-tuning technique without center wavelength shift. AB - We propose and experimentally demonstrate a simple and flexible scheme for a wavelength-spacing-tunable multichannel filter exploiting a sampled chirped fiber Bragg grating based on a symmetrical modification of the chirp ratio. Symmetrical bending along a sampled chirped fiber Bragg grating attached to a flexible cantilever beam induces a variation of the chirp ratio and a reflection chirp bandwidth of the grating without a center wavelength shift. Accordingly, the wavelength spacing of a sampled chirped fiber Bragg grating is continuously controlled by the reflection chirp bandwidth variation of the grating corresponding to the bending direction, which allows for realization of an effective wavelength-spacing-tunable multichannel filter. Based on the proposed technique, we achieve the continuous tunability of the wavelength spacing in a range from 1.51 to 6.11 nm, depending on the bending direction of the cantilever beam. PMID- 17130907 TI - Large-pitch kagome-structured hollow-core photonic crystal fiber. AB - We report the fabrication and characterization of a new type of hollow-core photonic crystal fiber based on large-pitch (approximately 12 microm) kagome lattice cladding. The optical characteristics of the 19-cell, 7-cell, and single cell core defect fibers include broad optical transmission bands covering the visible and near-IR parts of the spectrum with relatively low loss and low chromatic dispersion, no detectable surface modes and high confinement of light in the core. Various applications of such a novel fiber are also discussed, including gas sensing, quantum optics, and high harmonic generation. PMID- 17130909 TI - Asymmetric cryptography based on wavefront sensing. AB - A system of asymmetric cryptography based on wavefront sensing (ACWS) is proposed for the first time to our knowledge. One of the most significant features of the asymmetric cryptography is that a trapdoor one-way function is required and constructed by analogy to wavefront sensing, in which the public key may be derived from optical parameters, such as the wavelength or the focal length, while the private key may be obtained from a kind of regular point array. The ciphertext is generated by the encoded wavefront and represented with an irregular array. In such an ACWS system, the encryption key is not identical to the decryption key, which is another important feature of an asymmetric cryptographic system. The processes of asymmetric encryption and decryption are formulized mathematically and demonstrated with a set of numerical experiments. PMID- 17130910 TI - Fuzzy logic and maximum a posteriori-based image restoration for confocal microscopy. AB - We propose a maximum a posteriori image restoration approach to 3D confocal microscopy. The image field is suitably modeled as a Markov random field, resulting in a Gibbs distributed image. A fuzzy-logic-based potential is employed in the Gibbs prior. Unlike other potentials, the fuzzy potential distinguishes intensity variation due to genuine edges and noise. The proposed approach has generated artifact-free restored confocal microscopy images. PMID- 17130911 TI - Inverse scattering for high-resolution interferometric microscopy. AB - We extend the applicability of inverse scattering for optical coherence tomography (OCT) to the case of high numerical aperture focusing optics. We include the effects of tight focusing so that the approach is applicable to any interferometric microscopy method. The applicability to modalities, such as OCT and optical coherence microscopy, enables computed reconstruction of three dimensional volumes from en face temporal ranging data. Simulations show that the computed structure outside of the focal plane exhibits spatially invariant resolution on par with the resolution achieved at the focal plane. PMID- 17130912 TI - Least-squares fitting of carrier phase distribution by using a rational function in fringe projection profilometry [corrected]. AB - A least-squares fitting technique for the carrier phase component in fringe projection profilometry is presented. The carrier phase distribution for an arbitrary measurement system can be perfectly described with a rational function, whose coefficients can be estimated by least-squares fitting to the measured reference phases, so that the restrictions and limitations in the existing techniques are eliminated. PMID- 17130913 TI - Fabry-Perot etalons using colloidal photonic crystal mirrors. AB - Fabry-Perot etalons have been fabricated with 3D colloidal photonic crystal mirrors. The colloid films were optimized for high reflection and low loss to provide good finesse values in the 1200-1700 nm spectral range. A cavity quality factor of 2400 and a finesse of 8 together with sharp 0.5 nm wide resonance transmission peaks are reported that attest to the relatively good optical quality of the three-dimensionally structured films and the promise of self assembly colloidal crystal chemistry in providing novel microporous optical interferometers for potential applications such as environmental and biological sensing. PMID- 17130914 TI - Sweep optical frequency synthesizer with a distributed-Bragg-reflector laser injection locked by a single component of an optical frequency comb. AB - A sweep optical frequency synthesizer is demonstrated by using a frequency stabilized optical frequency comb and injection-locked distributed-Bragg reflector (DBR) laser diode. The injection-locked DBR laser acts as a single frequency filter and, simultaneously, a high-gain amplifier of the optical frequency comb. The frequency instability of the heterodyne beat signal between two independently injection-locked DBR lasers is measured to be 2.3 x 10(-16) at 1 s averaging time. The output frequency of the sweep optical frequency synthesizer can be precisely tuned over 1 GHz, and a saturated absorption spectrum of the Cs D2 line at 852 nm is recorded by the injected DBR laser. PMID- 17130915 TI - Phase-shifting point-diffraction interferometer developed by using the electro optic effect in ferroelectric crystals. AB - A novel and simple phase-shifting point-diffraction interferometer using a z-cut lithium niobate wafer is proposed. The pinhole is realized by an optical lithography process, aluminum deposition, and subsequent lift-off on the surface of the wafer. The phase shifting is obtained by inducing the electro-optic effect along the z crystal axis. We demonstrate experimentally the possibility of retrieving an aberrated wavefront. PMID- 17130916 TI - Lidar detection using a dual-frequency source. AB - A new technique of dual-frequency Doppler-lidar measurement is investigated. This technique is based on the use of a coherently locked, tunable, dual-frequency laser source and is shown to accurately measure velocities as small as 26 mum/s. It is generated by exploiting the nonlinear dynamics of a semiconductor laser through a proper combination of optical injection and operating conditions. PMID- 17130917 TI - Near common-path optical fiber interferometer for potentially fast on-line microscale-nanoscale surface measurement. AB - We introduce a new surface measurement method for potential online application. Compared with our previous research, the new design is a significant improvement. It also features high stability because it uses a near common-path configuration. The method should be of great benefit to advanced manufacturing, especially for quality and process control in ultraprecision manufacturing and on the production line. Proof-of-concept experiments have been successfully conducted by measuring the system repeatability and the displacements of a mirror surface. PMID- 17130918 TI - Low-loss subwavelength metal C-aperture waveguide. AB - We present a design of a linear optical waveguide that utilizes a C-shaped metallic nano-aperture that efficiently transports light while maintaining a spot size of lambda/10. The performance of a C-aperture waveguide is superior to both a regular ridge waveguide and other surface plasmon based metal nano-optical waveguides. The energy transport mechanisms are explained by the coupling of an aperture surface resonance and the thickness resonances inside the guide channel. Finite-difference time-domain simulations of gold C-aperture waveguides are performed for a 1.5 microm wavelength incident plane wave. The 1/e decay length in power transmission is predicted to be approximately 2.5 microm. The total power throughput is 1.66 for the 2.55 microm long guide, with an intensity 6 times that of the incident wave at a distance 120 nm from the exit plane, having a spot size of 150 nm. PMID- 17130919 TI - Modulation instability in silicon photonic nanowires. AB - We demonstrate that strong modulation instability (MI) of copropagating optical waves can be observed in Si photonic nanowires with a length of only a few millimeters. We consider two distinct cases, namely one in which one wave propagates in the normal group-velocity dispersion (GVD) region and the other one experiences anomalous GVD, and a second case in which both waves propagate in the anomalous GVD region. In both cases we show that, for comparable optical powers, the peak value of the MI gain spectrum is 2 to 3 orders of magnitude larger than that achieved in optical fibers. PMID- 17130920 TI - Multichip vertical-external-cavity surface-emitting lasers: a coherent power scaling scheme. AB - We propose an efficient coherent power scaling scheme, the multichip vertical external-cavity surface-emitting laser (VECSEL), in which the waste heat generated in the active region is distributed on multi-VECSEL chips such that the pump level at the thermal rollover is significantly increased. The advantages of this laser are discussed, and the development and demonstration of a two-chip VECSEL operating around 970 nm with over 19 W of output power is presented. PMID- 17130921 TI - Nanometer-scale ablation with a table-top soft x-ray laser. AB - Ablation of holes with diameters as small as 82 nm and very clean walls was obtained in poly(methyl methacrylate) focusing pulses from a Ne-like Ar 46.9 nm compact capillary-discharge laser with a freestanding Fresnel zone plate diffracting into third order. These results demonstrate the feasibility of using focused soft x-ray laser beams for the direct nanoscale patterning of materials and the development of new nanoprobes. PMID- 17130922 TI - 250 W end-pumped Nd:YAG laser with direct pumping into the upper laser level. AB - A high-power longitudinally pumped Nd:YAG laser using direct pumping into the upper laser level is demonstrated. With an absorbed pump power of 438 W an output power of 250 W was realized, which results in an optical-to-optical efficiency of 57%. To the best of our knowledge, this is the first demonstration of a high output power 885 nm pumped laser design. PMID- 17130923 TI - Unifying approach to left-handed material design. AB - We show that equivalent circuits offer a qualitative and even quantitative simple explanation for the behavior of various types of left-handed (or negative-index) metamaterials. This allows us to optimize design features and parameters while avoiding trial and error simulations or fabrications. In particular, we apply this unifying circuit approach in accounting for the features and in optimizing the structure employing parallel metallic bars on the two sides of a dielectric film. PMID- 17130924 TI - In vivo photoacoustic flow cytometry for monitoring of circulating single cancer cells and contrast agents. AB - A new photoacoustic flow cytometry was developed for real-time detection of circulating cells, nanoparticles, and contrast agents in vivo. Its capability, integrated with photothermal and optical clearing methods, was demonstrated using a near-infrared tunable laser to characterize the in vivo kinetics of Indocyanine Green alone and single cancer cells labeled with gold nanorods and Indocyanine Green in the vasculature of the mouse ear. In vivo applications are discussed, including selective nanophotothermolysis of metastatic squamous cells, label-free detection of melanoma cells, study of pharmokinetics, and immune response to apoptotic and necrotic cells, with potential translation to humans. The threshold sensitivity is estimated as one cancer cell in the background of 10(7) normal blood cells. PMID- 17130925 TI - Enhanced second-harmonic generation in AlGaAs/AlxOy tightly confining waveguides and resonant cavities. AB - We demonstrate second-harmonic generation (SHG) from sub-micrometer-sized AlGaAs/AlxOy artificially birefringent waveguides. The normalized conversion efficiency is the highest ever reported. We further enhanced the SHG using a waveguide-embedded cavity formed by dichroic mirrors. Resonant enhancements as high as approximately 10x were observed. Such devices could be potentially used as highly efficient, ultracompact frequency converters in integrated photonic circuits. PMID- 17130926 TI - 300 microJ noncollinear optical parametric amplifier in the visible at 1 kHz repetition rate. AB - We demonstrate an order-of-magnitude energy scaling of a white-light seeded noncollinear optical parametric amplifier in the visible. The generated pulses, tunable between 520 and 650 nm with sub-25-fs duration, had energies up to 310 microJ with 20% blue-pump-to-signal energy conversion efficiency at 540 nm. This new ultrafast source will make possible numerous extreme nonlinear optics applications. As a first application, we demonstrate the generation of tunable vacuum ultraviolet pulses. PMID- 17130927 TI - Green-pumped high-power optical parametric oscillator based on periodically poled MgO-doped stoichiometric LiTaO3. AB - A high-power 532 nm-pumped multikilohertz nanosecond optical parametric oscillator using a periodically poled 1.0 mol.% MgO-doped stoichiometric lithium tantalate crystal that could be operated from room temperature to 200 degrees C without damage is reported. A broad continuous tuning range from 855 to 1410 nm was achieved within a single domain period. Efficient operation of high peak power and watt level average power with a power conversion of 62.5% was measured. These results show that a high-resolution high average power visible tunable source can be realized. PMID- 17130928 TI - Design of achromatic phase shifters for spaceborne nulling interferometry. AB - The designs of various achromatic phase shifters (APSs) envisaged for nulling interferometry and the search for extrasolar planets or systems from space are discussed in this Letter. It is shown that APS designs can sensibly influence the performance achieved by a simple Bracewell interferometer if they introduce a field of view (FoV) reversal and a pupil flip along one interferometer arm. Analytical and numerical simulations are presented, showing severe signal-to noise ratio loss and a weak capacity for serendipitous discoveries. The use of FoV-reversal APS should therefore be restricted to the characterization of already known extrasolar planets rather than to their detection. PMID- 17130929 TI - Rigorous analysis of shadowing effects in blazed transmission gratings. AB - Blazed transmission gratings have become crucial components in many hybrid optical systems. Shadowing effects are known to occur at their passive blaze facets, which may impair the system's efficiency performance. For optical designs, it is desirable to have a simple but accurate description of this phenomenon. We show that the efficiency reduction in low diffraction orders is dominated by a linear dependence on the ratio of wavelength to grating period rather than a quadratic dependence as proposed in extended scalar theory. The strength of the electromagnetic shadowing will be determined using rigorous diffraction methods and discussed with respect to imaging optical components. Results are compared to existing ray-optical models. PMID- 17130931 TI - Gaussian beams and minimum diffraction. AB - I show that different legitimate measures of the amount of diffraction lead to contradictory conclusions concerning the beam profile that experiences minimum diffraction for a given beam width. In particular, when the Renyi entropy is used, the Gaussian beams no longer provide minimum diffraction, i.e., they do not have maximum quality. PMID- 17130930 TI - Permanent recording of light helicity on optically inactive metal surfaces. AB - We report on an unusual permanent recording of light helicity on optically achiral metals. Following a number of circularly polarized (CP) or elliptically polarized (EP) femtosecond laser pulses, well-defined periodic surface structures are found on metal surfaces. These surface structures show different orientation when formed by left CP/EP compared with right CP/EP light. The formation of these structures is attributed to the interference between the incident light and the excited surface plasmons. To our knowledge, this is the only phenomenon that can permanently record light helicity with an optically inactive material. PMID- 17130932 TI - Multi-dark-state resonances in cold multi-Zeeman-sublevel atoms. AB - We present our experimental and theoretical studies of multi-dark-state resonances (MDSRs) generated in a unique cold rubidium atomic system with only one coupling laser beam. Such MDSRs are caused by different transition strengths of the strong coupling beam connecting different Zeeman sublevels. Controlling the transparency windows in such an electromagnetically induced transparency system can have potential applications in multiwavelength optical communication and quantum information processing. PMID- 17130933 TI - Scattering of vacuum states by dynamic plasmon singularities: generating photons from vacuum. AB - A metallic nanoparticle surrounded by an amplifying medium results in a boundary condition that creates a singularity in the particle's dynamic polarizability at a critical value of the gain. When this boundary condition is time dependent, parametric excitation of the electromagnetic vacuum results in photon emission. Estimates of the vacuum emission from nanostructures embedded in high-gain laser dyes excited by femtosecond lasers predict energies nearly 2 orders of magnitude larger than the spontaneous emission background. PMID- 17130934 TI - Effects of surface roughness on reflection spectra obtained by terahertz time domain spectroscopy. AB - We present an analytical model that shows that reflection from a rough surface causes a Gaussian frequency roll-off for the spectral magnitude of a terahertz wave and reduces the signal-to-noise ratio of terahertz time-domain spectroscopy. The parameter that determines the width of the frequency roll-off is the standard deviation of the surface height distribution. Measurements of terahertz waves reflected from copper powder samples provide experimental evidence for this effect. PMID- 17130935 TI - Dual-pump coherent anti-Stokes-Raman scattering microscopy. AB - We introduce dual-pump coherent anti-Stokes-Raman scattering (dual-CARS) microscopy. This new technique permits simultaneous imaging of two species characterized by different molecular vibrations, as well as the removal of nonresonant background. This is achieved by using three synchronized laser pulses probing two different vibrations. We demonstrate the virtues of the method by imaging a mixture of nondeuterated and deuterated lipids, clearly distinguishing the individual components and their organization in the mixed arrangement. Further, dual-CARS images of lipid stores in living Caenorhabditis elegans nematodes show that the suppression of the nonresonant background results in significantly enhanced image contrast. PMID- 17130936 TI - Balanced optical-microwave phase detectors for optoelectronic phase-locked loops. AB - A balanced optical-microwave phase detector for the extraction of low-jitter, high-power, and drift-free microwave signals from optical pulse trains is presented. The phase detection is based on electro-optic sampling with a differentially biased Sagnac loop. Because the timing information is transferred in the optical domain, the regenerated microwave signal is robust against drifts and photodetector nonlinearities. In a first experimental implementation, 3 fs in loop relative timing jitter (integrated from 1 Hz to 10 MHz) between a 44 MHz optical pulse train and a 10.225 GHz microwave signal is demonstrated. PMID- 17130937 TI - Single attosecond pulses from high harmonics driven by self-compressed filaments. AB - We show that isolated subfemtosecond, extreme ultraviolet (XUV) pulses can be generated via harmonic generation in argon by few-cycle infrared pulses formed through filamentation-induced self-compression in neon. Our calculations show that the time structure of the XUV pulses depends sensitively on both the amplitude and the phase modulation that are induced in the driving pulse during the self-compression process. PMID- 17130938 TI - 35 J broadband femtosecond optical parametric chirped pulse amplification system. AB - We report on what is believed to be the first large-aperture and high-energy optical parametric chirped pulse amplification system. The system, based on a three-stage amplifier, shows 25% pump-to-signal conversion efficiency and amplification of the full 70 nm width of the seed spectrum. Pulse compression to 84 fs achieved after amplification indicates a potential of 300 TW pulse power for 35 J amplified pulse energy. PMID- 17130939 TI - Terahertz multiwavelength phase imaging without 2pi ambiguity. AB - We present a terahertz phase imaging method with multiwavelengths. This novel approach can image an object with a larger optical length compared with using the largest wavelength in the terahertz spectrum and does not involve the usual phase unwrapping in the detection of phase discontinuity. Furthermore, this technique can effectively reduce the noise background. We present two examples to demonstrate the validity of this new method. Both measurements show that multiwavelength phase imaging is a straightforward and efficient phase data processing method in terahertz imaging applications. PMID- 17130940 TI - [Epidemiological transition: the other side of the coin]. AB - The model of epidemiological transition proposed by Omram explains the changes in disease patterns in communities. In societies with a high level of development this model has been complemented with the study of the post transition process. In this context, the emergence of allergic diseases, asthma and subsequently, of autoimmune diseases, has reached worrisome proportions in some countries, and no model can explain these changes. The hygiene theory supported by Strachan gives a reasonable explanation to this phenomenon. It postulates that the reduction of early exposure to biological agents, along with an improvement of sanitation conditions, immunizations and medical therapies, causes an asymmetrical immunological response, favoring the expression of Th2 response. The hygiene theory does not fully explain by itself what is happening in developed countries and it is not universally accepted. Chile is experiencing an epidemiological transition from a high burden of infectious diseases to a growing prevalence of non communicable diseases. In a previous similar setting in developed countries, there is some evidence to suspect that asthma, allergic and autoimmune diseases are becoming part of the epidemiological situation of Chile. PMID- 17130941 TI - [Potential cost effectiveness of a rotavirus vaccine in Chile]. AB - BACKGROUND: Cost effectiveness studies are essential to assess the real value of interventions with preventive or therapeutic objectives. AIM: To assess the theoretical cost-effectiveness of a vaccine against rotavirus in Chilean children of less than five years of age. MATERIAL AND METHODS: An economic model was developed based on information on disease incidence, health care costs associated with treatment and the effectiveness and costs of vaccination. Net disease and vaccination costs were estimated from the health system perspective and were compared with life years and disability-adjusted life-years (DALYs) gained using a 3% discount rate. Local administrative and accounting hospital data and vaccine efficacy data were used to estimate healthcare costs and cost-effectiveness of vaccination. RESULTS: A rotavirus vaccination program would prevent 10 deaths due to rotavirus gastroenteritis, 6,245 related hospitalizations and 41,962 outpatient visits during the first five years of life, per vaccinated cohort. For every 1,000 children born, the healthcare service spends US$15,077 on treatment of gastroenteritis. From the healthcare perspective, vaccination would yield a cost-effectiveness ratio of US$11,261 per DALY when the price of the vaccine is US$24 per course. CONCLUSIONS: Rotavirus vaccine can effectively reduce the disease burden and healthcare costs of rotavirus gastroenteritis and can be a cost-effective investment compared to other options. PMID- 17130942 TI - [Innate immunity restoration in patients with HIV/AIDS infection associated with antiretroviral therapy]. AB - BACKGROUND: Highly active antiretroviral therapy (HAART) in HIV/AIDS infection induces an important reduction of the viral load (VL) and an immune system reconstitution. CD4+ T lymphocyte count is the immunological measurement commonly used for the follow up of HIV/AIDS patients. AIM: To study prospectively the restoration of the innate immune system in patients with HIV/AIDS infection during their first year on HAART. PATIENTS AND METHODS: 25 naive HIV/AIDS patients, from San Jose Hospital and University of Chile Clinical Hospital, Santiago, Chile, were studied between years 2002-2003. Every 4 months after HAART initiation, CD3+, CD4+, CD8+ T lymphocytes and CD16/56+ natural killer (NK) cells were quantified by flow cytometry. NK cell cytotoxicity was measured using radioactive chrome liberation (Cr51). Tumor necrosis factor alpha (TNF-alpha) and interleukin-10 (IL-10) were measured in peripheral blood mononuclear cells and viral load was determined using Amplicor HIV-1 from Roche Diagnostics Systems. RESULTS: Thirteen of the 25 patients continued in the study. They were all males, average age 35 years old (23-50). At baseline average CD4+ count was 146 cells/microL (31-362) and average viral load was 82.000 copies/mL (4.000 290.000). A raise in CD3+, CD4+, CD8+, and CD16/56 cells was noted at months 9-12 of therapy. Viral load became undetectable in the same period. NK cell function was decreased at the beginning of the therapy (1-4 months), reaching its highest values at months 9-12. There was no significant change in IL-10. TNF-alpha increased in six patients during the study. CONCLUSIONS: In this group of patients, innate immunity was restored during HAART. These results should be confirmed in studies with a longer follow up period and also measuring cytokines such as MIP-1a, MIP-1ss and RANTES. PMID- 17130943 TI - [Controlled trial of dexamethasone compared with droperidol and ondansetron for the treatment of postoperative nausea and vomiting]. AB - BACKGROUND: Dexamethasone is useful as prophylaxis for postoperative nausea and vomiting (PONV). AIM: To study the short term efficacy of dexamethasone to treat PONV in adults without prophylaxis, and compare its efficacy with that of droperidol and ondansetron. MATERIAL AND METHODS: A prospective study was performed with 120 consecutive adult patients presenting PONV in the postanesthesia care unit (PACU) at a University teaching hospital. During the occurrence of PONV, patients were randomized to receive in a double blind manner dexamethasone 8 mg IV (Group 1, n=40), droperidol 1.25 mg IV (Group 2, n=40), or ondansetron 2 mg IV (Group 3, n=40). Risk factors for PONV were recorded. Evaluations were made until discharge from the PACU and included presence of PONV, degree of sedation, and other potential adverse effects of the study drugs. Short term efficacy was defined as the percentage of patients free of PONV during all the stay in PACU after treatment. RESULTS: General data was similar for the 3 groups. Mean +/- SD stay in PACU after treatment was 101+/-34 minutes in Group 1, 93+/-33 minutes in Group 2, and 99+/-32 minutes in Group 3 (NS). Short term efficacy (CI 95%) was 55% (40-70%) in Group 1, 90% (81-99%) in Group 2, and 63% (48-78%) in Group 3 (p <0.05 for group 2 vs 1 and 3). There were no adverse effects attributable to the study drugs. CONCLUSIONS: Short term efficacy of dexamethasone to treat PONV was similar to ondansetron, but inferior to droperidol. Further studies are needed to define the duration of this effect of dexamethasone. PMID- 17130944 TI - [Changes in the patterns of disease after the epidemiological transition in health in Chile, 1950-2003]. AB - BACKGROUND: During the twentieth century there was a change in the pattern of diseases in Europe, with an increase in the incidence of allergies and autoimmune disorders, that paralleled a decrease of infectious conditions. The Hygiene hypothesis proposes that these phenomena are causally related. AIM: To evaluate the epidemiological changes of allergic, autoimmune, and infectious diseases in Chile between 1950 and 2003. MATERIAL AND METHODS: Search for the incidence and prevalence of these diseases in the national records published by the Ministry of Health, as well as through a systematic search of national literature using PubMed and Scielo as search engines. RESULTS: The annual incidence of tuberculosis, rheumatic fever, measles, and typhoid fever has progressively diminished in Chile since 1970. Figures for the national prevalence for asthma, rheumatoid arthritis, and type I diabetes are scarce and difficult to compare, but clearly show an increasing epidemiological trend in the last 20 years. CONCLUSIONS: The national figures suggest that, although the country has only recently gone through an epidemiological transition in health problems, there are detectable changes that show the same trends described in Europe. PMID- 17130946 TI - [Endoscopic biliary stents for the temporary management of choledocholithiasis]. AB - BACKGROUND: Endoscopic extraction of biliary tract stones is safe and effective. When the procedure is not successful, the use of a temporary stent can be a solution. AIM: To prospectively analyze the usefulness of endoscopic biliary stents in the temporary management of biliary obstruction due to choledocholithiasis. MATERIAL AND METHODS: Analysis of 51 consecutive patients (age range 21-88 years, 34 females) with common bile duct stones that, from January 1999 to December 2001, were subjected to an endoscopic insertion of a biliary stent. RESULTS: The indications for stent placement were a large stone in 40 patients (78%), the insecurity of a complete biliary tract cleaning in eight (16%) and technical difficulties in three (6%). Twenty seven patients (52.9%) were jaundiced and 17 (33.3%) had cholangitis. The prostheses remained in place until definitive resolution of the choledocholithiasis in 47 patients (92%) and migrated in 4 (8%). Bilirubin levels became normal in all cases with jaundice and infection resolved in all those with cholangitis. The definitive treatment of choledocholithiasis was done endoscopically in 28 patients (58%) and surgically in 20 (42%). Three patients were lost from follow up. Of these, one patient (2%) died 14 months later due to a recurrent cholangitis. The remaining two patients were asymptomatic and with the prostheses still in place 522 and 560 days post stent placement. CONCLUSIONS: When the immediate endoscopic resolution of choledocholithiasis is not possible, temporary stenting is a simple and safe therapeutic alternative that allows patients to be free of obstructive complications until the definitive treatment is carried out. PMID- 17130945 TI - [Quality of life of women depressed in the post-partum period]. AB - BACKGROUND: Depression is common among women, specially during breeding. AIM: To characterize post-partum depression in a group of women attending a primary health care clinic and its relationship to quality of life. MATERIAL AND METHODS: The sample included women meeting criteria for Major Depression, with a child of up to 11 months old. Exclusion criteria included the presence of psychosis, history of mania, alcohol abuse, illegal drug use, high suicide risk, and receiving mental health care in the last three months. Structured interviews used were the Edinburgh postpartum depression scale (EPDS), Mini International Neuropsychiatric Interview (MINI), Medical Outcomes Study Questionnaire (SF-36) and questions about incapacity and health care use. RESULTS: The sample included 159 women with a median age of 27 years (range: 16-43 years). Thirty three percent were married, 30.8% lived in common law marriage, 9.4% were divorced, and 26.4% were single. Most (89.3%) were housewives, 31% were students, and 6.9% were employed. The average score on the EPDS was 17 points (S.D. 4.2). The average SF 36 somatic score was 42.7 points (S.D. 8.2), and the emotional score was 30.3 (D.S. 0.3). The relation between the average score on the EPDS and the somatic and emotional scores was statistically significant (p=0.000-0.006). CONCLUSIONS: Most women were mildly to moderately depressed. Their depression was associated with a marked impairment of activities of daily living. These findings add to the evidence suggesting that depression is associated with marked disability cultures, even when the depression is mild to moderate. PMID- 17130947 TI - [Smoking interventions in primary health care: smoking profile of women and beliefs and attitudes of local health care teams]. AB - BACKGROUND: Chile has one of the highest prevalence rate of smoking in the world. Brief counseling interventions for smoking cessation at the primary health care level are effective. Compliance with counseling intervention is strongly associated with beliefs and attitudes of the primary health care team that deliver it. The effectiveness of these interventions improve if they are applied to smoking populations with higher motivation of change and high self-efficacy for quitting. AIM: To study the smoking profile of a group of smoking women in Santiago and to identify beliefs and attitudes of the primary health care team members to implement smoking cessation interventions. MATERIAL AND METHODS: A cross-sectional design that included 306 women smokers attending two primary health care clinics in Santiago. Perceptions, beliefs and attitudes of 34 primary care team members from three clinics in Santiago were explored using a qualitative methodology. RESULTS: The study identified a subgroup of 18% of women highly motivated to quit (decisional stage of change) and a 58% with a high self efficacy. Beliefs and attitudes of staff at the clinics were characterized by invisibility, ambivalence and fatalism regarding the effectiveness of smoking cessation interventions. CONCLUSIONS: There is a subgroup of smoking women with a high probability of quitting if they receive an appropriate counseling. Developing a systematic approach for smoking cessation intervention at the primary care setting in Chile should consider the invisibility, ambivalence and fatalism of primary health care team members towards this topic. PMID- 17130948 TI - [Differences among chronic restrained eaters: the influence of motivational systems]. AB - BACKGROUND: Restrained eaters (RE) are individuals who restrain their food intake on a regular basis as they are frightened to gain weight. However, they tend to overeat under conditions of anxiety. It has been shown that RE possess a behavioral inhibition system that is more active in tonic terms, which would partially explain their affective vulnerability. Even so, the influence of variations in the activation levels of the emotional systems on the eating behavior of a RE is still unknown. Our hypothesis is that variations of such systems will give place to two types of RE: a successful or a non-successful one. AIM: To assess the influence of variations on the activation of motivational systems in food intake of RE. MATERIALS AND METHODS: As part of a factorial experimental design, 105 undergraduate university students were part of an experimental test for inducting food intake. Then they reported their levels of dietary restraint and their emotional behavioral preferences. RESULTS: Differences in the activation of motivational systems were significantly related to differences in food intake (F= 7.210; p= 0.001). Additionally, food intake for those RE with a predominant inhibition system tended to be higher than for those with a more active approach system, though the latter did not reach a significant difference (F=0.718; p=0.399). CONCLUSIONS: Although more investigations are required, our data suggest that the success of retaining the diet among the RE would depend on their profile of affective reactivity (affective style). There are putative implications for research on anorexia and obesity. PMID- 17130949 TI - [Exercise-induced asthma as perceived by pediatric patients and their parents]. AB - BACKGROUND: Exercise is a frequent trigger of symptoms in asthmatic children and it worsens their quality of life. AIM: To compare the perception about exercise among asthmatic pediatric patients and their parents. MATERIAL AND METHODS: Asthmatic patients with symptoms related to exercise, were tested with an exercise challenge test following the Tal protocol. Before testing, a questionnaire about symptoms triggered by exercise was answered by children and their parents. The data was analyzed with a Kappa correlation test. RESULTS: Seventy five patients, aged 6 to 15 years, were studied. Forty one percent exercised less than one hour per week. Although 64% reported to experience respiratory difficulty and 80% cough during exertion, 87% were willing to perform more exercise. Forty percent of all patients had a positive challenge test for exercise-induced asthma. Correlation between patient's and parent's answers about the effect of physical activity exercise was low, with a kappa of 0.53. There was no correlation between exercise test and the answers to the questionnaire. CONCLUSIONS: Children with asthma frequently have exercise-associated symptoms and parental perception about this problem is very low. PMID- 17130950 TI - [Experience in the use of a new endocardial stimulation electrode]. AB - BACKGROUND: Since February 2002, Flextend electrode catheters are used at the Cardiovascular Unit of the Catholic University Clinical Hospital. These transvenous catheters have an IS-1 connector, silicone coating, active fixation and retractile helix with dexamethasone acetate. AIM: To report early and one year results using Flextend catheters. MATERIAL AND METHODS: Retrospective analysis of all patients that received an implant of a Flextend catheter in the Unit. RESULTS: Forty one Flextend catheters were implanted in 24 patients, without acute displacement or clinical pericarditis. In 18 electrode catheters located in the right atrium the mean values for p wave, stimulation threshold and impedance at the moment of placement, were 2.9+/-1.4 mV, 0.8+/-0.4 V and 522+/-86 Ohms, respectively. The figures 24 hours after placement were 3.6+/-2.1 mV, 0.8+/ 0.3 V and 612+/-69 Ohms, respectively. In 23 electrode catheters in the right ventricle, mean values at the moment of placement for R wave, stimulation threshold and impedance were 11.3+/-3.6 mV, 0.8+/-0.2 V and 756+/-108 Ohms, respectively. The figures 24 hours after placement were 3.6+/-2.1 mV, 0.8+/-0.3 V and 612+/-69 Ohms, respectively. In one patient, the stimulation threshold increased two months after placement and required a new intervention. CONCLUSIONS: Flextend catheters ca be placed successfully with a low rate of complications and stable function parameters on follow up. PMID- 17130951 TI - [Impact of air pollution by fine particulate matter (PM10) on daily mortality in Temuco, Chile]. AB - BACKGROUND: Temuco (304,000 inhabitants) has high levels of air pollution, mainly due to fine particulate matter of less than 10 microm (PM10). The effects of this pollution on population health have not been studied. AIM: To study the short term effects of PM10 on daily mortality in Temuco, in Southern Chile, due to respiratory and cardiovascular causes. MATERIAL AND METHODS: We followed the APHEA methodology (Air Pollution and Health European Approach) by estimating poisson multivariate regression models and controlling by trends, seasonality and meteorology. The PM10 variable was introduced after controlling by the confounders and checking by statistical adjustment and autocorrelation of errors. Mortality data was obtained from the Ministry of Health, registering age, gender, place of residence and cause of death. Cancer, respiratory and cardiovascular deaths, occurring between 1997 and 2002, were recorded for this study. RESULTS: There was a significant and positive association between PM10 concentration and daily mortality caused by respiratory disease (p-value=0.046, relative risk (RR) 1.236, 95% confidence interval (CI) 1.004-1.522) and cardiovascular diseases in people aged 65 years and more (p-value=0.042; RR 1.176 95% CI 1.006-1.374). CONCLUSIONS: There is a significant association between daily air pollution by PM10 particulate matter and mortality in Temuco, Chile. PMID- 17130952 TI - [White blood cell lysis syndrome after autologous peripheral blood stem cell transplantation in the treatment of renal AL amyloidosis. Case report]. AB - The treatment of AL amyloidosis was not successful until the advent of myeloablative chemotherapy consisting of high-dose intravenous melphalan followed by autologous peripheral blood stem cell transplantation. This new treatment has achieved better survival rates and, remarkably, it has obtained complete remission. Among patients with renal involvement, achievement of a complete hematological response was associated with a 50% reduction in proteinuria and stable creatinine clearance in more than 2/3 of patients. Despite of these excellent results, this new therapy is associated with significant toxicity, including the development of acute renal failure due to white blood cell lysis syndrome. We report a 59 year-old female with a nephrotic syndrome due to primary amyloidosis successfully treated autologous stem cell transplantation who developed acute renal failure caused by white blood cell lysis syndrome. The patient required treatment with granulocytic colony stimulating factor and intermittent hemofiltration and was discharged 23 days after melphalan administration with a satisfactory renal function and white blood cell count. After one year of follow up, she maintains a good glomerular filtration rate, a proteinuria of less than, 1 g/day and normal hematological values. PMID- 17130953 TI - [Permanent pacemaker implantation in patients with persistent left superior vena cava and absent right superior vena cava. Report of three cases]. AB - Persistent left superior vena cava and absent right superior vena cava is an uncommon anatomical association. This is a challenging situation for permanent pacemaker implantation. We report three patients with this anomaly and a permanent pacemaker successfully implanted through the left superior vena cava and coronary sinus, without acute or chronic complications. PMID- 17130954 TI - [Thyroid carcinoma as a cause of upper airway obstruction in children. Case report]. AB - Upper airway obstruction (UAO) can be a severe medical condition with a high mortality in children. We report a 10 year-old girl with UAO due to papillary thyroid carcinoma. The study confirmed a thyroid cancer. The patient was referred to our centre for the evaluation of dyspnea and hoarseness. She was admitted in severe respiratory distress. Her chest X-ray revealed a critical narrowing of the cervical trachea and extensive infiltration of the lung with a miliary pattern; CT scan revealed a thyroid mass with bilateral pulmonary dissemination. An early surgical approach with total thyroidectomy and tracheotomy was performed. The study revealed a thyroid carcinoma. The patient was then referred to a specialized centre to receive radioiodine treatment [corrected] Recognition of thyroid carcinoma in children requires a high suspicion index. An early CT scan and fiberoptic assessment could show UAO in many unsuspected lesions. PMID- 17130955 TI - [Hepatitis C in Chile: burden of the disease]. AB - Hepatitis C virus-associated chronic hepatitis is one of the most important causes of liver-related mortality and morbidity worldwide. This review analysis the available clinical and epidemiological information about this disease in Chile and compares it with data available from Latin America and other countries. Chronic hepatitis C seroprevalence in the general Chilean population is 1.15% by ELISA III and 0.85% by recombinant immunoblot assay (RIBA). Mortality due to cirrhosis (all causes) in Chile is one of the highest in the world. We show indirect evidence that chronic hepatitis C may account for a significant proportion of these deaths. The disease is the most common cause for liver transplantation in adults. Based on the available information, we conclude that chronic hepatitis C is an important cause of disease and mortality in Chile. PMID- 17130956 TI - [Treatment of neurocysticercosis: a review]. AB - Neurocysticercosis (NCC) is the most common parasitic disease of the central nervous system. Several drugs, such as drugs against tapeworms, praziquantel or albendazole associated to corticosteroids, have been tested for the treatment of this condition. Although some have claimed the reduction or involution of cystic or granulomatous lesions, there is no consensus about the efficacy of these treatments. The natural evolution of the disease is not clear and this hampers the assessment of treatment effects. Moreover, there are no good imaging or clinical indicators that can predict the progression or spontaneous resolution of lesions, specially at the meningeal or ventricular compartment. Therefore, evidence based medicine does not have a definitive answer about the treatment, neither of seizures, the most common manifestation of NCC, or the varied and complex meningeal and ventricular involvement. This review includes experts opinions to give the clinician some clues for decision making in the treatment of NCC. PMID- 17130957 TI - [Multiple pregnancies and folic acid]. PMID- 17130959 TI - [Conflicts of interest: an academic proposal]. PMID- 17130960 TI - [Antibiotic prophylaxis in neutropenic patients]. PMID- 17130961 TI - [What determines the time-lag for publications in Revista Medica de Chile?]. AB - The number of manuscripts submitted for publication in Revista Medica de Chile has increased steadily. Consequently, the number of external experts that generously contribute in the peer review process has also been enlarged. When the manuscripts return to the authors with specific requests to be improved, according to the criticisms raised by their reviewers or the editors, more time is added until a corrected version is received and accepted. After acceptance, the manuscripts are programmed by the editors to be included in future monthly issues of the Revista. The number of manuscripts that can be included in an issue is limited by the number of pages accorded between the publishers and the owner institution. Therefore, an additional time-lag occurs: research articles and case reports may be published 6-8 months after they were finally accepted, while reviews and special articles have to wait even more. As experienced in prestigious journals worldwide, a computational program to handle the editorial process through the Internet is needed, but it is not yet available in Spanish language. Although it should fasten communications and facilitate the authors a rapid insight in the stages of the editorial process where their manuscripts are, the increasing number of submissions, the current acceptance rate and the limitation in space will still retard the publications once they are accepted. PMID- 17130962 TI - [Self assessment of strengths, weaknesses and self confidence of primary care physicians taking care of rheumatic diseases]. AB - BACKGROUND: Rheumatologic diseases are common and frequently managed by primary care physicians. AIM: To assess strengths, weaknesses and self confidence of primary care physicians in the management of rheumatic diseases. MATERIAL AND METHODS: A self assessment and anonymous questionnaire was mailed to primary care physicians of two Chilean regions. Using a 10 points Likert scale, they were asked about personal interest, undergraduate training, continuous medical education, availability of medical literature, complementary laboratory tests and consultation with a rheumatologist. Medical skills, knowledge, therapeutic approach and performance of rheumatologic procedures were evaluated under the item confidence. RESULTS: Three hundred forty seven out of 763 physicians (45%) answered the questionnaire. Their age range extended from 25 to 75 years, 59% were male, 58% were Chilean and 74% worked in the Metropolitan region. The worst evaluated parameters were availability of literature with a score of 2.2+/-2, access to consultation with a rheumatologist (3.8+/-2.2) and to continuous medical education (4.3+/-2.7). Physicians had a better confidence in their knowledge (6.7+/-1.5) and in their therapeutic approach (6.1+/-1.5). The worst confidence score was for shoulder injection therapy (3.4+/-2.6). Continuous medical education correlated with knowledge but not with clinical skills. CONCLUSIONS: Primary care physicians perform a bad assessment of their skills in rheumatology. They have a low level of confidence in their clinical skills to perform rheumatologic procedures. Continuous medical education improves confidence in knowledge but not in skills. PMID- 17130963 TI - [Effectiveness of Leuprolide acetate therapy every three months in central precocious puberty]. AB - BACKGROUND: Precocious puberty may reduce final adult height, and affected children may suffer social and emotional problems. The efficacy of treatment with a long acting agonist analogue of the gonadotropin releasing hormone (aLHRH) has been well demonstrated. AIM: To evaluated the efficacy of a new formulation of aLHRH (leuprolide, Lupron) for the suppression of gonadotropin activation and clinical signs of puberty. MATERIAL AND METHODS: Eleven children (ten females) with idiopathic central precocious puberty, with a mean chronological age of 7.5+/-1.8 years and a bone age of 9.7+/-1.8 years were recruited. Testicular volume in the male was 15 ml. In females, Tanner stage for breast development was between 2-4 and mean ovarian volume was 2.3+/-0.8 ml. They were treated during 18 months with aLHRH, 11.25 mg administered intramuscularly every three months. RESULTS: Clinical, hormonal and ultrasonographic signs of puberty regressed in all patients. The degree of suppression of LH was 87.7+/-5.1% at the end of the 18 months. No significant changes in bone mineral content were observed during the treatment period. CONCLUSIONS: Leuprolide (aLHRH) 11.25 mg, injected every three months, is effective for the control of central precocious puberty and allows to reduce the number of yearly injections from 12 to 4. PMID- 17130964 TI - [Endovascular treatment of superior vena cava syndrome]. AB - BACKGROUND: Superior vena cava syndrome (SVCS) is caused by the obstruction of venous drainage from the upper portion of the body. Common clinical findings are headache and cervical, facial and upper limb edema. Occasionally, clouding of consciousness appears. AIM: to report our experience with endovascular treatment of SVCS. MATERIAL AND METHODS: Retrospective review of all patients with SVCS subjected to endovascular treatment between 1999 and 2005. RESULTS: Eight patients were treated, all of them with malignancies. Six had a benign obstruction due to the presence of a chemotherapy catheter located in the superior vena cava, one had obstruction secondary to radiation therapy and one a tumor compression of the superior vena cava. Two patients underwent thrombolytic therapy. Angioplasty and stenting was performed in all patients. The chemotherapy catheter was removed to all patients and installed again in one. One patient had a hemothorax secondary to a simultaneous needle lung biopsy under video thoracoscopy. No patient died in relation to the procedure. Congestive signs and symptoms subsided in all patients within 24 hours after the procedure. During follow up, only one patient had symptoms related to vena cava obstruction and three died due to their malignant tumor. CONCLUSIONS: Endovascular treatment of SVCS has a low rate of complications and provides immediate and mid-term symptom relief. PMID- 17130965 TI - [Molecular characterization of ABO*O alleles at the ABO group locus in three chilean populations]. AB - BACKGROUND: Among the allelic variants of blood groups, the molecular characterization of ABO blood group has clinical and anthropological importance. AIM: To perform a characterization of the molecular variants of the allele ABO*O of the ABO blood group. MATERIAL AND METHODS: Eighty four subjects of Aymara origin, living in Northern Chile, 75 individuals of Huilliche origin, living in Southern Chile and 82 subjects living in Santiago (Central Chile), were studied. All individuals were of group O, homozygotes for G261- deletion, that defines O1 alleles. Mutations G188A, G261-, G542A, T646A and C771T, described for alleles O1, O1variant and G542A were determined by PCR-RFLP (polymerase chain reaction restriction fragment lenght polymorphism). RESULTS: Allele O1variant has frequencies of 0.65, 0.81 and 0.6 in Aymara, Huilliche and Santiago subjects, respectively. The figures for allele O1 are 0.35, 0.19 and 0.4, respectively and those for the allele with G542A mutation are 0.119, 0.113 and 0.079, respectively. CONCLUSIONS: These results are concordant with the reported higher frequency of allele O1variant in South American aboriginal populations. The frequencies of G542A allele in these Chilean individuals are lower than those described for Amazon aborigines. PMID- 17130966 TI - [Hereditary colorectal cancer: Molecular analysis of APC and MLH1 genes]. AB - BACKGROUND: Among colorectal cancer hereditary variants, two syndromes show a predisposition to the disease based on germline mutations: Familial Adenomatous Polyposis (FAP) and Hereditary Nonpolyposis Colorectal Cancer (HNPCC). AIM: To screen mutations in FAP and HNPCC families in Chile. MATERIALS AND METHODS: Two FAP and one HNPCC families were studied. The APC gene (for FAP patients) and the MLH1 gene (for HNPCC patients), were screened for mutations on genomic DNA. The molecular analysis was performed through polymerase chain reaction, Single Strand Conformer Polymorphism (SSCP) and DNA sequencing. Mutations were defined as changes in the DNA sequence leading into a stop codon and a truncated protein. RESULTS: In the two FAP families the analysis revealed a mutation consisting in the deletion of five nucleotides named c.3927_3931delAAAGA. The genetic study of the HNPCC family demonstrated the insertion of one adenine in codon 168 of exon 6, named c.504insA. DISCUSSION: Germ-line mutations were identified in the three families. The relevance of these studies in a better knowledge of cancer susceptibility, and the possibility of identifying in relatives in risk by molecular diagnosis. PMID- 17130967 TI - [Perioperative risk among morbid obese patients subjected to gastric bypass]. AB - BACKGROUND: Bariatric surgery is a complex procedure not exempt of complications. AIM: To assess mortality and complications of excisional gastric bypass among morbidly obese subjects. MATERIAL AND METHODS: Prospective analysis of 684 morbid obese patients (age range 14-70 years, 525 females) subjected to an excisional gastric bypass. Major postoperative complications and mortality were registered. RESULTS: Mean body mass index (BMI) of the subjects was 43.7 kg/m2. One hundred sixty two patients had a BMI between 35 and 39.9 kg/m2, 419 had a BMI between 40 and 49.9 kg/m2 and 103 had a BMI over 50 kg/m2. Two patients with a BMI of 52 and 56 kg/m2 respectively, died in the postoperative period (0.3%). Thirty six patients had major complications. Anastomotic fistula was the most common complication in 12 patients (1.7%). Fourteen patients required a new operation due to complications. None of these died. The mean operative volume of the surgical team was 124 patients per year. CONCLUSIONS: Excisional gastric bypass has a low rate of mortality and complications, if the surgical team operates a large volume of patients. PMID- 17130968 TI - [Short-term Lycopersicum esculentum consumption may increase plasma high density lipoproteins and decrease oxidative stress]. AB - BACKGROUND: Tomato has a high antioxidant capacity due to its high content of vitamin C, vitamin E and lycopene that is a powerful free radical scavenger. However, the effects of tomato on plasma lipoproteins is not well known, and there is little evidence about the relationship between tomato consumption and oxidative state changes in humans. AIM: To assess in vivo the effects of dietary supplementation with pure concentrated tomato juice on short term changes in oxidative state and plasma lipoproteins in healthy volunteers. SUBJECTS AND METHODS: Seventeen healthy volunteers were studied. They received a supplement of pure tomato juice during 7 days. At baseline, at the end of the supplementarion period and eight days after the end of the supplementation, a blood sample was drawn to measure total antioxidant capacity (TRAP), enzymatic antioxidants (catalase and superoxide dismutase), non-enzimatic antioxidants (lycopene and a tocopherol) and plasma lipoproteins. RESULTS: Lycopene level increased early and significantly in comparison with basal levels (48%; p < 0.05). TRAP, catalase and superoxide dismutase did not change significantly. HDL cholesterol increased significantly in 5.6+/-4.3 mg/dL (p < 0.002) on the second sampling period, improving the ratio cholesterol/HDL. It returned to baseline in the third period. CONCLUSIONS: Dietary supplementation of concentrated tomato juice significantly increases lycopene levels and HDL cholesterol. Non significant changes observed in TRAP, catalase and superoxide dismutase were observed during the supplementation period. PMID- 17130969 TI - [Prevalence of Bartonella henselae antibodies in Chilean children, adolescents and veterinary workers]. AB - BACKGROUND: Bartonella henselae is the causative agent of cat-scratch disease. AIM: To study the seroepidemiology of Bartonella henselae in healthy Chilean children and in a population with occupational risk. MATERIAL AND METHODS: Serum IgG antibodies were determined by indirect fluorescence technique in 181 children and adolescents and in 107 technical and professional workers involved in the care of cats. Samples with titers equal to or greater than 64 were considered positive. RESULTS: Twenty four (13.3%) children and 11 (10.3%) occupational risk subjects were seropositive. No significant differences by age and gender were observed. CONCLUSIONS: Assuming that seroprevalence indicates level of exposure to Bartonella henselae, these results suggest that this infection is endemic in Chile and, for this reason, the best antibody titer to diagnose acute cat-scratch disease should be higher than the figure recommended by the Centers for Disease Control in the in United States. PMID- 17130970 TI - [Rapid identification of non tuberculous mycobacteria by restriction pattern analysis]. AB - BACKGROUND: The frequency of diseases caused by non tuberculous mycobacteria has increased in the last years. Their clinical diagnosis is difficult, mainly in immunocompromised patients. The identification of these mycobacteria by traditional methods is based on phenotypic characteristics and the results are obtained two to four weeks after their isolation in primary cultures. AIM: To report a new identification method for non tuberculous mycobacteria. MATERIAL AND METHODS: The restriction pattern analysis method was implemented. It is based on the amplification, using polymerase chain reaction (PCR), of a polymorphic region of 440 base pairs that codifies Hsp65 protein, followed by a digestion with BstE II and Hae III restriction enzymes. The results were compared with patterns established for each strain. RESULTS: Sixty four strains of mycobacteria obtained from clinical samples and seven reference mycobacteria, were identified using the traditional methods and restriction pattern analysis. The latter method identified the same strain as the former in 87.5% of cases. In the remainder 12.5% of cases there was no agreement between both methods. In these, the sequencing of a fragment of a gene that codifies 16S ribosomal RNA, confirmed the correct identification by restriction patterns. CONCLUSIONS: Restriction pattern analysis is a rapid identification method for non tuberculous mycobacterial strains. PMID- 17130971 TI - [Effectiveness of infliximab in patients with Behcet syndrome and severe uveoretinitis. Report of five cases]. AB - The efficacy and safety of repeated administration of infliximab was evaluated in five patients (two men, three women) with Behcet syndrome accompanied by severe uveoretinitis. Ocular and extra ocular inflammation was suppressed in all patients during the observation period without any serious adverse reactions. The results in these patients suggests that TNF-a blockade is effective in patients with severe ocular Behcet syndrome. PMID- 17130972 TI - [Diagnosis of acute disseminated encephalomyelitis using a stereotaxic biopsy: Report of one case]. AB - We report a 15 year-old female presenting with behavioral disturbances, headache, left hemiparesis and paresis of the vertical gaze. CAT scan and magnetic resonance showed an involvement of right thalamus, third ventricle and medial temporal lobe suggesting an encephalitis or lymphoma. 201Thalium SPECT suggested a lymphoma. A stereotaxic biopsy showed a subacute demyelinizing lesion, compatible with an acute disseminated encephalomyelitis. The patient was treated with Methylprednisolone with resolution of symptoms. She remains in good condition after one year of follow-up. PMID- 17130973 TI - [Loss of biventricular resynchronization due to an uncommon cause: Report of one case]. AB - Cardiac resynchronization therapy is a non-pharmacological treatment for patients with dilated cardiomyophaty and congestive heart failure. The success of this therapy depends of permanent biventricular stimulation. We report an 84 year-old man, with intermittent loss of biventricular pacemaker stimulation despite having adequate sensing and stimulation thresholds in the right atrium and both ventricles. The problem was solved after correcting some programming parameters. PMID- 17130974 TI - [Levodopa for Parkinson's disease: What have we learned?]. AB - Parkinson's disease (PD) is the second most common neurodegenerative disease in the world. Dopamine deficit is the cornerstone of its clinical manifestations. Levodopa, the main treatment for this condition, was first used for PD more than 40 years ago and today it still is the most powerful treatment for this disease. In recent years many advances have been made for understanding of the neurochemical mechanisms of this drug. Furthermore, new insights about the genesis of motor complications secondary to its use are known, specially related with the mode of its administration. This article updates the pharmacology of levodopa and its implications for the pathophysiology and treatment of PD. The new available presentations of levodopa are also reviewed. The implications of these advances for the treatment of this disease are commented. PMID- 17130975 TI - [Update in the diagnosis and therapy for pulmonary arterial hypertension]. AB - Pulmonary Arterial Hypertension includes a heterogeneous group of disorders with a common genetic, pathological and hemodinamyc origin. It is characterized by a high pulmonary artery pressure due to a primary vascular disease, as a consequence of genetic and environmental factors. The common pathway is a vascular imbalance towards vasoconstriction and proliferation inside the small vessels. According to the World Health Organization, 2003, Pulmonary Arterial Hypertension is classified as idiopathic, familiar or associated to connective tissue diseases, HIV, drugs, porto-pulmonary hypertension, congenital intracardiac shunts and others. The diagnosis is based in hemodynamics. Echocardiogram is a non invasive and right ventricular catheterization is an invasive diagnostic tool. Follow up is based on a clinical and functional assessment through functional class classification, dyspnea scores and 6-minute walking test. The prognosis is historically devastating but new therapies are changing the natural history of the disease. New treatments have demonstrated improvement in symptoms, hemodynamic profiles and survival. Intravenous, subcutaneous or inhaled prostanoids such as Epoprostenol, Treprostinil or Iloprost respectively have been approved for Pulmonary Arterial Hypertension treatment as well as oral endothelial receptor blockers. They are all considered first line treatments for arterial pulmonary hypertensive patients with even better benefits than lung transplantation. Phosphodiesterase inhibitors (Sildenafil), have been recently approved for the treatment of pulmonary arterial hypertension. PMID- 17130976 TI - [Role of 18Fluorine-deoxyglucose (FDG) positron emission tomography in the management of lymphomas]. AB - Positron emission tomography (PET) with fluorine-18 labeled deoxyglucose (FDG) is useful in the management of lymphomas. In this review, some general concepts of this metabolic test are defined. It has an excellent diagnostic yield in Hodgkin disease as well as in most non Hodgkin lymphomas. Staging, restaging residual mass evaluation and the control of therapy are the main indications for FDG-PET. Images with FDG have a high diagnostic and prognostic value, that is superior to anatomical images and conventional staging techniques. They are also helpful for the assessment of tumor activity in abnormal lymph nodes or large masses that have been treated and reduce their size slowly or show an incomplete resolution. Currently, the resolution of dedicated PET equipments is 6 mm and bigger lesions can easily be detected. The main differences and advantages of FDG versus gallium 67 in lymphoma are also discussed, as well as the initial local experience with the technique in lymphoma patients. PMID- 17130977 TI - [Methodological quality of an article on the treatment of gastric cancer adopted as protocol by some Chilean hospitals]. AB - BACKGROUND: Surgery is a curative treatment for gastric cancer (GC). As relapse is frequent, adjuvant therapies such as postoperative chemo radiotherapy have been tried. In Chile, some hospitals adopted Macdonald's study as a protocol for the treatment of GC. AIM: To determine methodological quality and internal and external validity of the Macdonald study. MATERIAL AND METHOD: Three instruments were applied that assess methodological quality. A critical appraisal was done and the internal and external validity of the methodological quality was analyzed with two scales: MINCIR (Methodology and Research in Surgery), valid for therapy studies and CONSORT (Consolidated Standards of Reporting Trials), valid for randomized controlled trials (RCT). Guides and scales were applied by 5 researchers with training in clinical epidemiology. RESULTS: The reader's guide verified that the Macdonald study was not directed to answer a clearly defined question. There was random assignment, but the method used is not described and the patients were not considered until the end of the study (36% of the group with surgery plus chemo radiotherapy did not complete treatment). MINCIR scale confirmed a multicentric RCT, not blinded, with an unclear randomized sequence, erroneous sample size estimation, vague objectives and no exclusion criteria. CONSORT system proved the lack of working hypothesis and specific objectives as well as an absence of exclusion criteria and identification of the primary variable, an imprecise estimation of sample size, ambiguities in the randomization process, no blinding, an absence of statistical adjustment and the omission of a subgroup analysis. CONCLUSION: The instruments applied demonstrated methodological shortcomings that compromise the internal and external validity of the. PMID- 17130978 TI - [Controversies in the editorial process and in scientific publications]. PMID- 17130980 TI - [Undergraduate medical students' expectative of their desired profile as medical doctors]. AB - BACKGROUND: During the last few years, multiple new medical schools have emerged in Chile, associated to the constant preoccupation to provide a good quality medical care. This created the need to evaluate medical training programs and to open a discussion about the attributes that a good physician should have. AIM: To evaluate the medical student's perception of the ideal medical doctor profile. MATERIAL AND METHODS: An analytical, descriptive and cross sectional study was designed. Eleven second year, 11 third year, nine fourth year, 13 fifth year, 6 sixth year and 8 seventh year students were studied. Data collection was gathered by focus groups. Codes and triangulation were used for data analysis. RESULTS: As attitudes and moral-ethical values, students valued the absence of discrimination a listening attitude and empathy. Among job related issues, they valued responsibility and punctuality. Emotional and legal self-care were valued as self related attitudes. Among skills, competences and capacities, a value was given to communicational skills, team work and professional easiness. Knowledge about medical and non medical topics was appraised. The valued attributes among duties and activities were patient diagnosis, treatment and education, team leadership and continuous medical training. CONCLUSIONS: These findings should help to design new curricula for medical schools. PMID- 17130981 TI - [Student patient relationship from the patient's point of view]. AB - BACKGROUND: Patients are becoming increasingly active in their relationship with medical professionals. Their relationship with medical students needing to learn clinical skills, may be specially problematic if patients are not willing to accept their involvement in the medical team. AIM: To examine patient's perceptions of their relation with medical students and their agreement to let students be part of the treating team. MATERIAL AND METHODS: Qualitative study using taped semi-structured interviews addressed to inpatients from one public and one private hospital in Chile. RESULTS: Both groups of patients acknowledged that students dedicated more time to them, but they expressed their preference to limit student's participation to clinical history taking and physical examination. They also expected them to be observers rather than actors. Patients from the private hospital emphasized that only one student per instructor should participate in their care. Patients from the public hospital were more compliant about student's participation. The right to refuse students' involvement in their care was clearly known by all patients from the private system and by most patients from the public hospital. CONCLUSIONS: Patients in Chilean public and private hospitals were in general positive regarding student's participation in their care. Students' clinical practice ought to strictly respect patients's rights, and patients should be considered volunteers who generously agree to cooperate with the education of medical students. PMID- 17130982 TI - [Presence of antineutrophil cytoplasmic antibodies (ANCA) and anti Saccharomyces cerevisiae antibodies (ASCA) among patients with ulcerative colitis]. AB - BACKGROUND: The diagnosis of inflammatory bowel disease is supported by clinical findings and complementary tests. The presence of specific serological markers could be helpful in the characterization of this condition. AIM: To assess the prevalence of ANCA and ASCA in a group of patients with ulcerative colitis (UC) and its association with clinical features. MATERIAL AND METHODS: Sixty four patients with UC in remission (age range 16-72 years, 33 males) were studied. In a venous blood sample ANCA were measured by indirect immunofluorescence and ASCA by enzyme immune assays for IgG and IgA. RESULTS: Forty four percent of patients were positive for ANCA, 9% for ASCA and 6% for both markers. There was a significant correlation between the presence of ANCA and duration of the UC (<5 years 50%, 5-10 years 42.9%, 15 years 30%) and the number of crises (one crises 31%, 2-5 crises 51.9% and >5 crises 87.5). The proportion of colectomized patients with positive ANCA was higher (57.1%). CONCLUSIONS: The prevalence of ANCA in the studied population is similar to the published data. The presence of ANCA was significantly higher in UC patients with shorter evolution, higher number of crises and in those with a history of colectomy. There was a low prevalence of ASCA positive patients. PMID- 17130983 TI - [Report of a new mutation in CYBB gene in two patients with X linked chronic granulomatous disease]. AB - BACKGROUND: The X-linked form of chronic granulomatous disease (CGD) is a primary immunodeficiency that affects phagocytes of the innate immune system and is characterized by an increased susceptibility to severe bacterial and fungal infections. It is caused by mutations in the CYBB gene, which encodes the 91-kD subunit of phagocyte NADPH oxidase. AIM: To identify the mutation in the CYBB gene in two unrelated patients from Chile with the diagnosis of X-linked CGD and their families. PATIENTS AND METHODS: The molecular genetic defects of two unrelated patients from Chile with X-linked CGD caused by defects in the CYBB gene were investigated. The underlying mutation was investigated by single strand conformation polymorphism (SSCP) analysis of PCR-amplified genomic DNA and by sequencing of the affected gene region. RESULTS: We found an insertion c.1267_1268insA in exon 10 leading to a frameshift mutation. This mutation is a novel report. We also identified a splice site mutation in the other patient, that presented a c.1326 +1 G>A substitution in intron 10. The mutation was also detectable in his heterozygous mother. CONCLUSIONS: This is the first report of the clinical and molecular characterization of Chilean patients with mutations in CYBB gene. PMID- 17130984 TI - [Program of eating disorders in Chilean adolescents and young adults: six months outcome]. AB - BACKGROUND: Eating disorders may have serious organic consequences derived from under nutrition, specific nutrient deficiencies and electrolytic disturbances and reach a mortality as high as 12%. AIM: To describe the features and outcome after six months of treatment of patients attending the Eating Disorders Unit at the Catholic University Hospital in Chile. MATERIAL AND METHODS: Review of medical records of patients with eating disorders that received a multidisciplinary treatment by a team of nutritionists, psychiatrists and psychologists and were seen at least twice in a period of six months. RESULTS: The records of 81 patients (mean age 16.3+/-3 years, only one male) were included in the review. Forty nine patients had Anorexia Nervosa of the restricting type, five of the purging type, 22 had Bulimia Nervosa of the purging type and two of the non purging type and three patients had an eating disorder not otherwise specified. Eighty percent consulted within the first year of symptom appearance. Sixty five percent came from families with both parents living together, 57% had a rigid and agglutinated family structure and 65% had occult crises. Depression or dysthymia was found in 45% of patients and drug therapy was required at the beginning of treatment in 25%. Obsessive traits (40.4%) were significantly related to restrictive eaters. A past history of obesity or overweight was common. After six months of treatment, body mass index increasing significantly in restrictive eaters with obsessive traits and occult family crises was noticed. CONCLUSIONS: A multidisciplinary therapeutic intervention improved body mass index and family symptoms in patients with restrictive eating disorders. PMID- 17130985 TI - [Genotypying of clinical isolates of Helicobacter pylori by cagA, vacA and babA2 virulence associated genes. First detection of a babA2 positive strain in Chilean patients]. AB - BACKGROUND: Helicobacter pylori-associated gastroduodenal diseases depends on host characteristics, environmental conditions and bacterial virulence factors, such as cagA, vacA y babA2 gene products. Moreover, peptic ulcer disease has been related with cagA+, vacAs1m1 strains, while metaplasia and gastric cancer has been associated to cagA+, vacAs1 and babA2+ H pylori strains. Gene babA2 has not yet been described in clinical isolates from Chilean patients. AIM: To investigate the presence of cagA, vacA (s and m) and babA2 genes in clinical isolates of H pylori from Chilean patients. MATERIAL AND METHODS: Sixty six isolates from 41 patients were genotyped by PCR, using primers for s1a, s1b, s2, m1, m2, cagA and babA2 genes as previously described. RESULTS: cagA gene was detected in 16 isolates (24.2%) while vacAs1a, vacAs1b, vacAs2, vacAm1 and vacAm2 were detected in 28 (42.4%), 14 (21.2%), 17 (25.8%), 21 (31.8%) and 29 isolates (43.9%), respectively. One isolate (1.5%) was babA2 positive, being the first isolate with this genotype described in Chile. Besides the babA2+ genotype this clinical isolate also presented cagA+ and vacAs1a which has been related with metaplasia or gastric cancer. Five isolates showed an ulcerogenic profile cagA+, vacAs1m1. CONCLUSIONS: The results presented indicate the prevalence of vacAs1m1 genotype among the clinical isolates analyzed, and a low frequency of babA2 genotype. PMID- 17130986 TI - [Results of treatment for acromegaly in 53 patients: it is time of intervention]. AB - BACKGROUND: The treatment of choice for acromegaly is surgery that, according to the literature, is curative in 91% of pituitary microadenomas and 73% of macroadenomas. AIM: To report the results of surgical treatment in 53 patients with acromegaly. MATERIAL AND METHODS: Retrospective review of medical records of all patients with acromegaly, operated between 1984 and 2004. When necessary, patients were contacted by telephone to complete information or to perform biochemical or imaging studies. A normal value of insulin like growth factor I (IGF-1) for age and sex, a growth hormone (GH) nadir of less than 1 ng/ml during a glucose tolerance test or a basal GH of less than 2.5 ng/ml, all assessed three months after surgery, were considered as criteria for cure. RESULTS: Biochemical cure was achieved in 67% of patients with pituitary microadenomas and 21% of patients with macroadenomas. In 47% of patients with neuro-ophtalmological involvement, a partial or total recovery in the visual field defect was achieved. The most common surgical complications were transient diabetes insipidus in 19%, persistent diabetes insipidus in 4% and cerebrospinal fluid fistula in 4%. A lower size of the tumor and lower preoperative growth hormone levels were associated with a better chance of cure. CONCLUSIONS: The cure rates obtained in this group of patients are clearly lower than those reported abroad. These results stress the importance of having a national registry of acromegaly and the need to train neurosurgeons in the treatment of pituitary tumors. PMID- 17130987 TI - [Screening colonoscopy among first degree relatives of patients with colorectal carcinoma]. AB - BACKGROUND: First degree relatives of patients with colorectal carcinoma are at a higher risk of having the disease than the general population. Therefore, they should be subjected to screening colonoscopy. AIM: To assess the effectiveness of colonoscopy among first degree relatives of patients with colorectal carcinoma. MATERIAL AND METHODS: A free colonoscopy was offered to first degree relatives of patients operated on for colorectal cancer between 1998 and 2000. As inclusion criteria, subjects had to be asymptomatic, older than 40 years or less than 10 years younger than the index case. Each subject was contacted twice, inviting him/her to have a colonoscopy performed. RESULTS: Two hundred forty three relatives were contacted for the study and in 76, a colonoscopy was performed. Among the latter, a neoplasm was found in 13 (17%): One adenocarcinoma and 12 adenomas. Three of these lesions were located in the right colon. The main reason given by the 176 subjects that did not agree to have a colonoscopy was lack of interest. CONCLUSIONS: Screening colonoscopy is effective to detect adenoma and adenocarcinoma among first degree relatives of patients with colorectal carcinoma, however only 31% of all potential relatives agreed to undergo a colonoscopy. PMID- 17130988 TI - [Immunohistochemical expression of the E-cadherin-catenin complex in gastric cancer]. AB - BACKGROUND: The E-cadherin/catenin complex plays an essential role in the control of epithelial differentiation. Abnormal expression in tumors correlates with histological grade, advanced stage and poor prognosis. AIM: To evaluate the expression pattern of E-cadherin/catenin complex in gastric carcinoma and analyze their association with tumor clinicopathological features and patient survival. MATERIAL AND METHODS: Inmunohistochemical staining of E-cadherin, alpha and ss catenin was performed from paraffin specimens of 65 gastric carcinomas. RESULTS: Abnormal expression of E-cadherin, alpha and ss-catenin was demonstrated in 82%, 85% and 88% of gastric carcinomas, respectively. There was a significant correlation between abnormal expression and Lauren pathological classification and depth of infiltration, but not with tumor stage, positive lymph node metastases and survival. CONCLUSION: Abnormal expression of E-cadherin, alpha and ss-catenin occurs frequently in gastric carcinoma and correlates with histological grade. PMID- 17130989 TI - [Brain and spinal cord magnetic resonance imaging in spastic paraparesis associated to human T-lymphotropic virus]. AB - BACKGROUND: The spastic paraparesis associated to HTLV-1 causes degenerative pyramidal tract lesions of the spinal cord and affects cortical-nuclear connections in the brain. AIM: To report the findings of magnetic resonance imaging in patients with spastic paraparesis. MATERIAL AND METHODS: A magnetic resonance imaging of the brain and spinal cord was performed in 30 patients (24 females), mean age and evolution of 56 and 12 years respectively, with a clinical and virological diagnosis of tropical spastic paraparesis/HTLV-1 associated myelopathy (TSP/HAM). RESULTS: No patient had abnormal signals in the spinal cord parenchyma. However, an atrophy of the dorsal segment was observed in 87% of patients. Patients with the highest degree of atrophy showed a higher degree of functional impairment. Eleven patients had spinal cord conus atrophy, associated to neurogenic bladder or impotency. In 80% of patients, hyperintense subcortical white matter images in DP, T2 and Flair, mostly bi frontal, were detected. In half of them, small rounded and isolated images were observed. In the other half, eight or more images, generally larger and occasionally confluent, were found. Ten of 12 patients with confluent brain lesions showed different degrees of cognitive impairment. No patient had lesions in the corpus callosus, periventricular white matter, pons, medulla oblongata or cerebellum. CONCLUSIONS: Most patients with tropical spastic paraparesis have alterations in brain or spinal cord magnetic resonance imaging. The magnetic resonance lesions are concordant with functional impairment. The characteristics of the imaging in TSP/HAM patients can be helpful in the differential diagnosis of patients with paraparesis. PMID- 17130990 TI - [Ventricular assist device as a bridge to transplant in patients with cardiogenic shock. Preliminary experience in Chile with ABIOMED BVS 5000]. AB - Hospitalization and death due to heart failure and cardiogenic shock is frequent and currently is increasing among the adult population. Although cardiac transplantation is the most effective treatment in patients with end-stage heart failure, its availability is limited. While waiting for transplantation, some patients become refractory to treatment and deteriorate progressively. Secondary multi-organ damage could highly compromise the transplant success and also could contraindicate it. Mechanical ventricular assist devices allow reestablishing normal cardiac output and they have been used as a bridge to recovery and transplantation. We report four patients that underwent mechanical ventricular support using the ABIOMED BVS 5000 system as a bridge for transplantation. Two patients were connected to biventricular assistance; a third patient was connected to a left ventricular support and the fourth to a right ventricular support. Three were successfully transplanted and one died of refractory non cardiogenic shock. There were no complications related to the support system, such as infection, hemorrhage or stroke. In our experience, the ABIOMED BVS 5000 was an effective strategy as a bridge to heart transplant in patients in cardiogenic shock. PMID- 17130991 TI - [Endoluminal stent-graft placement for acute dissection of the descending aorta complicated with rupture. Report of one case]. AB - Acute dissection of the aorta, although not common, has early and highly lethal complications. The type A dissection is treated with surgery. Patients with type B dissections are treated with surgery if they have complications like rupture, growth or visceral ischemia. Surgery, however, has complications such as spinal cord ischemia. Endovascular grafts have less mortality and complications. We report a 59 years old male patient with a type B dissection complicated with rupture. He was treated successfully with the placement of an endoluminal graft. He was discharged five days after the procedure in good conditions. After one year of follow up, the patient remains asymptomatic. PMID- 17130992 TI - [Streptococcus constellatusas a causative agent of empyema. Report of one case]. AB - Streptococcus constellatus is a commensal microorganism in man but may cause infections in different locations. We report a 59 years old male with severe sequelae of a previous cerebrovascular accident that consulted in the emergency room for fever of 15 days of evolution. A right empyema was diagnosed. The bacteriological culture of the effusion disclosed the presence of Streptococcus constellatus and two anaerobic strains (Prevotella intermedia and Fusobacterium urealyticus). The patient was treated with a pleural drainage and received ceftriaxone and clindamycin during six weeks. He was discharged in good conditions and is asymptomatic after eight months of follow up. PMID- 17130993 TI - [Gut barrier in the critically ill patient: facts and trends]. AB - The disturbances of gut barrier in critically ill patients may influence their outcome and prognosis. Experiments in animals show that fasting and stress collaborate to produce intestinal atrophy and translocation of microorganisms and toxins. This fact is one of the main arguments to promote the use of early enteral feeding in critically ill patients. However, the intestinal barrier behaves differently in humans than in animals. The human enteral cells have a good tolerance to fasting and stress, mucosal atrophy is mild and it is not always associated with changes in intestinal permeability. Moreover, the relationship between intestinal permeability with sepsis and bacterial translocation is controversial. This last phenomenon also happens in normal subjects and may be a mechanism to build immunological memory. One of the most important factors that influence bacterial translocation is the microorganism, that under stress conditions can adhere to the intestinal cell and penetrate the intestinal barrier. Splanchnic ischemia and reperfusion is one of the main pathogenic factors in the failure of intestinal barrier. Finally, the fact that the small bowel is an inflammatory target of extra intestinal injuries, explains several clinical situations. The pathophysiology of the intestinal barrier definitely requires more research. PMID- 17130994 TI - [Wine, fibrinolysis and health]. AB - Cardiovascular diseases are the leading cause of death in both men and women in the world. Epidemiological and experimental studies have associated moderate wine consumption (1 to 2 glasses/day) with a decrease in cardiovascular diseases. This decrease is probably due to the effect of ethanol and polyphenols present in the wine. The cardioprotective benefit of wine may be due, in part, to a modulation of the expression of proteins involved in fibrinolysis. Endothelial cells (ECs) play a major role in maintaining normal hemostasis, regulating the balance between the synthesis and interaction of proteins that promote clot formation (thrombosis) and fibrinolytic proteins that facilitate clot lysis. These cells are a major site of synthesis of fibrinolytic proteins, such as tissue type plasminogen activator (t-PA), urokinase-type plasminogen activator (u-PA) and the major inhibitor/regulator of fibrinolysis, PAI-1. EC-mediated fibrinolysis is regulated and localized to the EC surface through specific receptors for u-PA, t PA and plasminogen. Evidence indicates that ethanol and polyphenols present in wine increase EC localized fibrinolisis. Upregulation of t-PA and u-PA activity and downregulation of PAI-1 may account, at least in part, for this net increase in fibrinolytic activity. The purpose of this review is to cover the main molecular and physiological aspects of moderate wine consumption mediated increase in fibrinolysis and reduction in cardiovascular risk. PMID- 17130995 TI - [Recommendations for the nutritional assessment of critically ill patients]. AB - Undernutrition affects 30 to 50% of hospitalized patients. The identification of patients with undernutrition or in risk to become undernourished is of utmost importance to prescribe a timely nutritional support. This article performs a critical review of anthropometrical, biochemical and functional parameters used for nutritional assessment. Prognostic indexes and other parameters used for the assessment of nutritional risk are also discussed. Some tools are proposed for the diagnosis of hospital malnutrition. PMID- 17130996 TI - [Availability of physicians in Chile at the year 2004]. AB - BACKGROUND: The number of physicians available in a given country, their efficiency, quality and specialization is of utmost epidemiological importance. AIM: To evaluate the availability of physicians in Chile. MATERIAL AND METHODS: The information about the number of physicians in Chile up to the year 2004, was obtained from the Ministry of Health, national universities and the register of immigrant physicians since 1950. RESULTS: The total number of physicians licensed to practice was 25,542, of whom 2,700 are immigrants. The physician/inhabitant ratio increased from 1/921 in 1998 to 1/612 in 2004. The greater impact in the increment of available physicians was given by the immigration of professionals and by the increase in the number of physicians graduated from national universities, mainly from the new private universities. Forty two percent of physicians work at public services and 61% of these are certified specialists. The regional distribution of general practitioners and basic specialists is adequate. Along the country, the mean physician/beneficiary ratio is 8.45/10,000, the specialist/beneficiary ratio is 4.9/10,000 and the general practitioner/beneficiary ratio is 2.3/10,000. CONCLUSIONS: The national information of available physicians, especially in the private sector, should be improved. Immigration of physicians should be regulated, maintaining validation examinations and a National Medical Test to assess medical proficiency should be instituted. PMID- 17130997 TI - [Valsalva: much more than a simple maneuver. Antonio Maria Valsalva (1666-1723)]. AB - Valsalva is a term commonly used to designate a special maneuver, consisting in a forced expiratory effort against a closed airway. We use Valsalva's maneuver many times in our clinical practice and we usually do not think about its origin. Most people do not know that this was originally described by an outstanding anatomist, surgeon and pathologist, Antonio Maria Valsalva, the mentor of another great anatomist, Giovanni Battista Morgagni. Valsalva's contribution was vast and description of Valsalva's maneuver was only a small fraction of the legacy that he left us, almost three hundreds years ago. We render a tribute to a great man, and learn about his life and work. PMID- 17130998 TI - [Clinical guidelines in ischemic vascular strokes, in Chile]. PMID- 17131001 TI - Characteristics and etiology of conjunctivochalasis: historical perspective. PMID- 17130999 TI - [Preliminary validation of a smelling test in a sample of Chilean population and its role in the differential diagnosis of Parkinson s disease]. PMID- 17131002 TI - The corneal epithelial stem cell niche. AB - In recent years, it has become generally accepted that the corneal epithelial stem cells are localized in the basal cell layer of the limbal epithelium. However, a number of questions remain regarding the number, markers, generation, and maintenance of the corneal epithelial stem cells. One of the key questions concerns what makes up the microenvironment or niche that is responsible for allowing the stem cells to remain and function throughout the life of the tissue. This review will consider the unique aspects of the limbus and compare these to what is known about other stem cell niches. PMID- 17131003 TI - Effects of topical antiglaucoma medications on the ocular surface. AB - The side effects of topical antiglaucoma medications and their preservatives range from ocular discomfort to sight-threatening alterations of the ocular surface. Conjunctival hyperemia, decreased tear production and function, and superficial punctate keratitis are among the most common signs seen on routine clinical examination. Squamous cell metaplasia and changes in cell morphology have been demonstrated by impression cytology studies and evaluation of biopsy specimens, and inflammatory effects are documented by the presence of inflammatory markers. The adverse effects of topical antiglaucoma eyedrops interfere with the treatment of glaucoma on two levels: first, the discomfort produced by the eye drops discourages patient compliance; and, second, long-term treatment with eyedrops is associated with a higher failure of filtration surgery. The detailed mechanism of inflammatory response and/or direct toxicity of eye drops has yet to be determined, but it may vary with the different classes of eye drops, different preservatives, and durations of treatments. Upcoming multicenter trials for new antiglaucoma eye drops should specifically evaluate ocular surface effects. PMID- 17131004 TI - Ocular rosacea. AB - Rosacea is a common skin disease that frequently involves the eye. Although the pathogenesis of the disease remains undefined, recent findings suggest that an altered inflammatory response plays an important role in both cutaneous and ocular rosacea. Ocular manifestations include lid and ocular surface alterations. Chronic inflammation can lead to corneal vascularization, which may compromise vision. Treatment of ocular rosacea is aimed at preventing irritation of the ocular surface (e.g., lubricants, lid hygiene) and controlling inflammation with topical and systemic anti-inflammatory drugs. Systemic tetracyclines are the mainstay of treatment. These drugs act multifactorially by decreasing bacterial flora and the expression of matrix metalloproteinases, altering meibum secretion, inhibiting the production of bacterial lipases, and providing an immunomodulatory effect. PMID- 17131005 TI - Kentucky Lions Eye Center/University of Louisville: an ocular surface center in development. PMID- 17131006 TI - What does it mean when a company says that a product is "approvable"? PMID- 17131009 TI - Crocodile tears. PMID- 17131010 TI - Bacterial biofilms and ocular infections. AB - Serious bacterial infections of the eye are often associated with abiotic prosthetic materials, such as contact lenses, intraocular lenses, and scleral buckles. In recent years, microbiologists have recognized substantial differences between bacteria growing in a sessile community attached to a surface and free living or planktonic bacteria. These sessile surface-attached communities are known as biofilms, whose properties have important consequences for clinical medicine. As a population, bacteria in biofilms are more resistant to antimicrobial agents and the immune system, and they are more persistent than planktonic bacteria in hostile environments. These characteristics are likely the result of both biofilm-specific phenotypes and increased phenotypic diversity within biofilms as compared with planktonic communities of bacteria. Bacterial biofilm formation has been observed on human tissues, as well as on abiotic prosthetic devices. A better understanding of biofilm formation may lead to the development of novel antimicrobial agents as well as prosthetic devices that are resistant to bacterial colonization. PMID- 17131012 TI - Assessment of tear film dynamics: quantification approach. AB - The dynamics of the tear film are reviewed with specific reference to the biophysical aspects: distribution, turnover and elimination through evaporation, drainage, and absorption. The review concentrates on quantitative assessments and is confined to aspects of the dynamics that can be fully and directly measured. The techniques of fluorophotometry, fluorescein clearance, lacrimal scintigraphy, evaporimetry and osmometry are described. Reports in the literature for values of tear turnover (flow), evaporation and osmolarity for normal and dry eyes are collated. Indices of tear film dynamics based on these measurements, including tear function index, total tear flow, and osmolarity, are discussed in relation to their potential in the differential diagnosis of dry eye and new referent values for the disease suggested. The limitations of derivation and application of these indices are discussed. PMID- 17131013 TI - New insights into the diagnosis and treatment of neurotrophic keratopathy. AB - Abnormalities of the ocular surface can be acquired or inherited disorders of the central nervous system. Loss of neural stimulation from the sensory division of the trigeminal nerve or from the autonomic nervous system can have devastating consequences on corneal epithelial wound healing and the precorneal tear film, leading to decreased vision. The pathogenesis of neurotrophic keratopathy is reviewed, and treatment modalities are recommended. PMID- 17131014 TI - The Ocular Surface Center at Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, Paris. PMID- 17131017 TI - Pilocarpine and tear secretion. PMID- 17131018 TI - In vitro alternatives to the use of animals in ocular toxicology testing. AB - Chemical substances, including household products, industrial chemicals, and cosmetics, must be tested for ocular toxicity or irritancy so that the public can be assured of their safety or warned of dangers associated with their use. The in vivo Draize test is the standard method used to meet this requirement; however, this test is coming under increasing criticism on scientific and ethical grounds. This has led to the development of a large number of proposed in vitro tests, some of which are routinely used to screen chemicals in toxicology laboratories. This review addresses regulations governing ocular irritancy testing and the current status of the movement toward use of alternative methods. Such methods include the use of cultured cells, hen's eggs, isolated animal eyes and corneas, human corneal epithelial cell lines, and the recently developed in vitro corneal equivalent models. The protocols for these methods are outlined, and their endpoints are described with respect to prediction of in vivo responses. The tests are evaluated in the context of the outcomes of validation studies and acceptance by regulatory agencies. While several of these tests yield useful information concerning ocular irritancy, to date, no in vitro alternative test has been validated as a replacement for the Draize test. If the goal of replacing the in vivo test while protecting the public from chemical eye injury is to be achieved, further development and improvement of alternative tests, as well as establishment of a human ocular toxicity data base, are required. PMID- 17131019 TI - Methodologies for the study of ocular surface disease. AB - The ability to obtain reliable results from clinical trials of therapies for ocular allergic disease and dry eye disease is often limited because of inadequate control of variables, such as environment, patient life style, compliance, and individual fluctuations that occur from one assessment visit to another. The controlled allergen challenge (CAC) model of allergic conjunctivitis allows signs and symptoms of the disease to be elicited in a physiologically accurate and reproducible manner. The rigid criteria for subject selection, the controlled allergic reaction, and the standardized and quantified grading systems allow for a reproducible baseline from which statistically and clinically significant differences between formulations can be assessed. Similarly, the controlled adverse environment (CAE) model for dry eye mimics the environmental stimuli that lead to ocular surface drying. Preselected subjects have a reproducible, homogeneous baseline reaction from which the effects of various treatments can be significantly evaluated and compared. CAC and CAE provide accurate means to study highly variable and individual ocular surface disease. PMID- 17131021 TI - Should you be a clinical investigator for a new pharmaceutical? PMID- 17131020 TI - Genetics of corneal disease for the ocular surface clinician. AB - Advances in the understanding of inherited corneal and external diseases may allow interventions that prevent the substantial vision impairment currently caused by these diseases. The observant clinician may first recognize inherited corneal and external diseases based on clinical examination and a careful family history. Researchers using positional cloning and candidate gene techniques have identified several disease-causing genes. Identification of the genes responsible for inherited corneal and external diseases will lead to more definitive diagnoses and represent the first step in development of effective therapies. Future endeavors are directed toward identifying additional inherited corneal and external diseases, the genes that cause them, and possible gene therapies to improve visual outcomes. PMID- 17131023 TI - Peter R. Laibson, MD. PMID- 17131025 TI - Etymology of the term "tear". PMID- 17131026 TI - Mucosal immunity and self-tolerance in the ocular surface system. AB - This paper articulates a new working hypothesis that explains many of the pathophysiological conditions described under the common rubric "dry eye" as altered states of mucosal immune regulation. A central principle of mucosal immune physiology is that the parenchymal tissues at the effector sites, i.e., the sites at which secretory antibodies are produced, maintain local signaling milieus that support differentiation of IgA+ plasmablasts and survival of IgA+ plasmacytes. These local signaling milieus also support robust regulatory networks that maintain tolerance to commensual microbes, benign antigens, and parenchymal autoantigens. The regulatory networks are mediated by cycles of interactions between successive generations of dendritic cells, which normally mature with tolerogenic functions, and regulatory T cells, which normally reinforce the system's ability to generate new tolerogenic dendritic cells. The systemic endocrine environment controls expression of the local signaling milieu in the mammary gland and in the prostate and male urethral glands. Emerging evidence indicates that the local signaling milieu in the lacrimal gland also is determined, in part, by the systemic endocrine environment. This working hypothesis suggests explanations for the excess incidence of Sjogren syndrome among women and for the mechanisms of several different immunophysiological states in addition to Sjogren syndrome that, like Sjogren syndrome, are associated with the classical symptoms and signs of dry eye. It also comprises a promising rationale for specific new approaches to therapy. PMID- 17131028 TI - Neurotrophic mediators and corneal wound healing. AB - Neurotrophic keratopathy is an ocular pathological condition that remains difficult to treat. The loss of trigeminal nerve function and corneal sensation that underlies this condition can lead to the development of various disorders of the cornea. Substance P, a sensory neurotransmitter produced by the trigeminal nerve, has been investigated for its effect on corneal epithelial wound healing. Substance P by itself has no direct effect on corneal epithelial migration, but it manifests a synergistic action with insulin-like growth factor-1 (IGF-1) in both epithelial migration in vitro and corneal wound healing in vivo. The minimal amino acid sequences of both substance P and IGF-1 that are required for such effects have been determined. With use of these minimal amino acid sequences, the potential adverse consequences of treatment with the full-length polypeptides may be avoided. The application of eye drops containing a substance P-derived peptide and IGF-1 has proved clinically effective for the treatment of patients with persistent epithelial defects of the cornea. PMID- 17131029 TI - Chronic ocular surface disease after allogeneic bone marrow transplantation. AB - Graft-versus-host disease (GVHD) is a common complication of allogeneic bone marrow transplantation (allo-BMT). Ocular surface disease (OSD) is one of the most common manifestations of chronic ocular GVHD, yet little is known about it. In this article, we review the available literature on this condition and present results from our study of the manifestations of OSD in the chronic phase (>3 months duration) post allo-BMT. Our study consisted of a retrospective chart review of 62 allo-BMT patients with chronic OSD evaluated at our center between 1995 and 2002. The clinical features, systemic associations, treatment, and status of OSD at the last follow-up are presented and discussed in the context of other reports of OSD in GVHD. PMID- 17131030 TI - Is the world flat for pharmaceuticals? PMID- 17131031 TI - Young investigators. PMID- 17131032 TI - Resonance assignments for the two N-terminal RNA recognition motifs (RRM) of the S. cerevisiae pre-mRNA processing protein Prp24. PMID- 17131033 TI - 1H, 13C and 15N resonance assignments of the pyrazinamidase from Mycobacterium tuberculosis. PMID- 17131034 TI - NMR assignment of the dengue 4 virus envelope protein domain III. PMID- 17131035 TI - Backbone resonance assignments of Ezrin C ERMAD in a non-covalent complex with Ezrin N FERM. PMID- 17131037 TI - A case of limited cutaneous systemic sclerosis developing anti-mitochondria antibody positive primary biliary cirrhosis after acute myocardial infarction. AB - In this report, we present a 63-year-old woman who had limited cutaneous systemic sclerosis and subsequently developed typical primary biliary cirrhosis after an acute myocardial infarction. The patient initially developed Raynaud's phenomenon, and 4 years later visited the clinic in 1994 complaining of abdominal distress, xerostomia, and xerophthalmia. A diagnosis of limited cutaneous systemic sclerosis was based on Raynaud's phenomenon, sclerodactyly and anti centromere antibodies. She was also found to have anti-inositol 1,4,5 trisphosphate receptor 3 (IP(3)R3) antibodies, but anti-mitochondrial antibodies were only weakly positive. Seven years later, she developed vertigo and nausea, and was hospitalized due to complaints of an oppressive sensation of the anterior chest. Electrocardiogram results showed a reduction of R waves and ST segment elevation in II, III, and aVf leads. Coronary angiography showed 99% obstruction of the left anterior descending artery and 50% of stenosis of the right coronary artery. Three years later, the patient was noted to have anti-mitochondrial antibodies. Retrospective analysis of the patient's sera showed that IP(3)R3 antibodies were decreasing. Since myocardium is particularly rich in mitochondria, it is thought that myocardial infarction may have been the triggering event that initiated antigen-presenting cells to selectively induce an anti-mitochondrial antibody response. PMID- 17131038 TI - Polymorphisms of the DNA polymerase beta gene in breast cancer. AB - DNA polymerase beta (Polbeta) provides most of the gap-filling synthesis at apurinic/apyrimidine sites of damaged DNA in the base excision repair pathway. Mutations in the gene encoding DNA polbeta have been identified in various carcinomas. We performed a case-control study to test the association between two polymorphisms in the polbeta gene: a Pro --> Arg change at codon 242 (the Pro242Arg polymorphism) and a Lys --> Met change at codon 289 (the Lys289Met polymorphism) and breast cancer risk and cancer progression. Genotypes were determined in DNA from peripheral blood lymphocytes of 150 breast cancer patients and 150 cancer-free, age-matched women (controls) by PCR-RFLP. A strong association between breast cancer occurrence and the Met/Met phenotype of the Lys289Met polymorphism [odds ratio (OR) 3.67; 95% confidence interval (CI) 1.87 7.56] and the Pro/Arg phenotype of the Pro242Lys polymorphism (OR 1.96; 95% CI 1.15-3.34) was found. Polymorphism-polymorphism interaction between the Met/Met phenotype of the Lys289Met and the Pro/Arg phenotype of the Pro242Arg variants increased the risk of breast cancer (OR 3.05; 95% CI 1.31-7.09). We did not observe any correlation between studied polymorphisms and breast cancer progression evaluated by node-metastasis, tumor size and Bloom-Richardson grading. In conclusion, Polbeta may play a role in the breast carcinogenesis and the Lys289Met polymorphism of the polbeta gene may be considered as an independent, early, molecular diagnostic marker in breast cancer. The Pro242Arg polymorphism may contribute to the carcinogenesis through the interaction with the Lys289Met and therefore may be regarded as a dependent, auxiliary marker. PMID- 17131039 TI - A novel BRCA1 mutation in an Indian family with hereditary breast/ovarian cancer. AB - Germ-line mutations in BRCA1 gene contribute to a majority of familial breast and ovarian cancers. A group of 23 Tamil Nadu (south India) patients with positive family history for breast and ovarian cancer were screened for BRCA1 mutations by conformation sensitive gel electrophoresis (CSGE) followed by sequencing. In the present study, we report a novel 1307delT mutation in exon 11 of BRCA1 gene in a 43-year-old woman of Indian origin with breast cancer. This mutation gives rise to a premature stop codon at amino acid residue 409 and also creates a novel DdeI restriction site. The same mutation was also detected in the patient's maternal uncle and his son through extended family analysis. The 1307delT is a novel mutation that has not been documented in any population or published report to the best of our knowledge. Identification of this novel mutation stresses the need for developing a database of BRCA1 mutations, which will aid in breast cancer screening in this population. PMID- 17131040 TI - Alterations of beta3-adrenoceptors expression and their myocardial functional effects in physiological model of chronic exercise-induced cardiac hypertrophy. AB - Physical training induces cardiovascular autonomic nervous system regulation adaptations, which could result from beta adrenergic receptor (AR) modifications. Among them, beta(3 )AR alterations have been recently reported but their functional effect remained to discuss. To explain the beta(3) AR gene expression in relation to function, we simultaneously studied the left ventricle (LV) beta(3) AR mRNA and protein levels and the myocardial functional effects of a beta(3) AR agonist following physical training. Forty rats were assigned to either a control (C; N = 20) or a trained (T; N = 20) group. The treadmill running protocol was performed for 8 weeks. Histological measurements on LV slices were quantified. The beta(3) AR mRNA abundance was studied with RT-PCR and beta(3) AR protein density with Western-Blot analysis. Myocardial functional effects of a beta(3) AR agonist, BRL37344 (10(-8) M), were studied in Langendorff perfused hearts. Histological data confirmed the adapted patterns of the physiological cardiac hypertrophy observed in T (P < 0.01), with a significant increase in arteries density (P < 0.01) and an unchanged collagen concentration. The beta(3) AR protein density was increased in T (154 +/- 38% in T vs. 100 +/- 24% in C; P < 0.05), but no change was noted concerning the beta(3) AR mRNA level. After BRL37344 perfusion LVDP, +dP/dT and -dP/dT, in C (P < 0.01), and only +dP/dT in T (P < 0.05) were decreased. Moreover, all LV hemodynamic parameters were more altered after BRL37344 in C than in T (P < 0.01).Thus, in this physiological model of cardiac hypertrophy, an increase of beta(3) AR density without beta(3) AR mRNA alteration was observed. Classical negative myocardial lusitropic and inotropic effects induced by a specific agonist of beta(3) AR were diminished in trained rats. PMID- 17131041 TI - Arc1p is required for cytoplasmic confinement of synthetases and tRNA. AB - In yeast, Arc1p interacts with ScMetRS and ScGluRS and operates as a tRNA Interacting Factor (tIF) in trans of these two synthetases. Its N-terminal domain (N-Arc1p) binds the two synthetases and its C-terminal domain is an EMAPII-like domain organized around an OB-fold-based tIF. ARC1 is not an essential gene but its deletion (arc1- cells) is accompanied by a growth retardation phenotype. Here, we show that expression of N-Arc1p or of C-Arc1p alone palliates the growth defect of arc1- cells, and that bacterial Trbp111 or human p43, two proteins containing EMAPII-like domains, also improve the growth of an arc1- strain. The synthetic lethality of an arc1-los1- strain can be complemented with either ARC1 or LOS1. Expression of N-Arc1p or C-Arc1p alone does not complement an arc1-los1- phenotype, but coexpression of the two domains does. Our data demonstrate that Trbp111 or p43 may replace C-Arc1p to complement an arc1-los1- strain. The two functional domains of Arc1p (N-Arc1p and C-Arc1p) are required to get rid of the synthetic lethal phenotype but do not need to be physically linked. To get some clues to the discrete functions of N-Arc1p and C-Arc1p, we targeted ScMetRS or tIF domains to the nuclear compartment and analyzed their cellular localization by using GFP fusions, and their ability to sustain growth. Our results are consistent with a model according to which Arc1p is a bifunctional protein involved in the subcellular localization of ScMetRS and ScGluRS via its N terminal domain and of tRNA via its C-terminal domain. PMID- 17131042 TI - Curcumin attenuates glutamate-induced HT22 cell death by suppressing MAP kinase signaling. AB - Glutamate induces cell death by upsetting the cellular redox homeostasis, termed oxidative glutamate toxicity, in a mouse hippocampal cell line, HT22. Extracellular signal-regulated kinases (ERK) 1/2 are known key players in this process. Here we characterized the roles of both MAP kinases and cell cycle regulators in mediating oxidative glutamate toxicity and the neuroprotective mechanisms of curcumin in HT22 cells. c-Jun N-terminal kinase (JNK) and p38 kinase were activated during the glutamate-induced HT22 cell death, but at a later stage than ERK activation. Treatment with a JNK inhibitor, SP600125, or a p38 kinase inhibitor, SB203580, partly attenuated this cell death. Curcumin, a natural inhibitor of JNK signaling, protected the HT22 cells from glutamate induced death at nanomolar concentrations more efficiently than SP600125. These doses of curcumin affected neither the level of intracellular glutathione nor the level of reactive oxygen species, but inactivated JNK and p38 significantly. Moreover, curcumin markedly upregulated a cell-cycle inhibitory protein, p21cip1, and downregulated cyclin D1 levels, which might help the cell death prevention. Our results suggest that curcumin has a neuroprotective effect against oxidative glutamate toxicity by inhibiting MAP kinase signaling and influencing cell-cycle regulation. PMID- 17131043 TI - Progesterone-dependent and -independent expression of the multidrug resistance type I gene in porcine granulosa cells. AB - A primary role of plasma membrane P-glycoprotein (P-gp), encoded by multidrug resistance type I (MDR1), is to protect against naturally occurring xenotoxics. Progesterone (P(4)) profoundly influences MDR1 expression in granulosa cells and luteal cells. Here, P(4) regulation of MDR1 expression was investigated in porcine granulosa cells using the P(4)-mediated promoter activity assay and a P4 receptor (PR) antagonist (RU-486). The promoter activity was measured chronologically for 48 h in cells transfected with the PR response element containing pGL3. LH could stimulate the promoter activity through endogenous P4, with a maximum activity at 5 h. MDR1 mRNA level was highly maintained at 24-36 h. Conversely, exogenous P4 prolonged the promoter activity to further 10 h, and the high level of MDR1 mRNA was maintained even at 48 h. RU-486 completely inhibited the promoter activity, but the level of MDR1 mRNA rapidly increased in the presence of RU-486. The granulosa cells may become susceptible to RU-486 as a xenotoxic to rapidly express MDR1 for protection against it. These results indicate that MDR1 is expressed in porcine granulosa cells through P4-dependent and -independent regulations. PMID- 17131044 TI - Mechanism of reduced colonic contractility in experimental colitis: role of sarcoplasmic reticulum pump isoform-2. AB - Inflammatory bowel diseases (IBD) such as Crohn's disease and ulcerative colitis are inflammatory disorders associated with decreased colonic contractility. Here we show that, in experimental colitis in rat induced by trinitrobenzenesulfonic acid, there is a decrease in contraction in response to carbamoylcholine and the sarco/endoplasmic reticulum Ca(+2) (SERCA) pump inhibitor thapsigargin. However, the decrease in contractility may occur due to decrease in the SERCA pump levels or their inactivation. Therefore, we examined the protein and mRNA levels for SERCA2 isoform, which is predominant isoform in colonic smooth muscle. There was a decrease in the levels of SERCA2 protein and mRNA levels in inflamed colonic muscle. These findings suggest that decreased SERCA pump levels is responsible for a decrease in the Ca(+2) stores in the sarco/endoplasmic reticulum that causes a decrease in the contractility in colonic smooth muscle leading to poor bowel movements. PMID- 17131045 TI - Direct evidence that two cysteines in the dopamine transporter form a disulfide bond. AB - We have generated a fully functional dopamine transporter (DAT) mutant (dmDATx7) with all cysteines removed except the two cysteines in extracellular loop 2 (EL2). Random mutagenesis at either or both EL2 cysteines did not produce any functional transporter mutants, suggesting that the two cysteines cannot be replaced by any other amino acids. The cysteine-specific reagent MTSEA-biotin labeled dmDATx7 only after a DTT treatment which reduces disulfide bond. Since there are no other cysteines in dmDATx7, the MTSEA-biotin labeling must be on the EL2 cysteines made available by the DTT treatment. This result provides the first direct evidence that the EL2 cysteines form a disulfide bond. Interestingly, the DTT treatment had little effect on transport activity suggesting that the disulfide bond is not necessary for the uptake function of DAT. Our results and previous results are consistent with the notion that the disulfide bond between EL2 cysteines is required for DAT biosynthesis and/or its delivery to the cell surface. PMID- 17131046 TI - Protoplast formation of the coccolithophorid Pleurochrysis haptonemofera in hypoosmotic K+ solution: shedding of the coccosphere and regrowth of the protoplast in normal medium. AB - Both coccolith-bearing cells (C-cells) and naked cells (N-cells) of the coccolithophorid Pleurochrysis haptonemofera can grow in salinities of more than 7 per thousand (about 20% of a "normal" sea water salinity [35 per thousand]), with the highest growth rates in salinities of more than 14 per thousand. Microscopic observations of cells suspended in 100 mM NaCl (7 per thousand) showed that, while N-cells were swelling uniformly all over the cell surface, C cells were bulging the plasma membrane from the hole of the coccosphere at the apical (flagellar) pole of the cell. Effects of several cations and anions on the morphological change of C-cells under hypoosmotic pressure were investigated. When 100 mM K(+) was used, protoplasts were released from the coccosphere completely in almost all the cells. This phenomenon was shown with K(+) most effectively. The protoplasts could grow in the fresh medium and form the first coccolith within 9 h. PMID- 17131047 TI - High photobiological hydrogen production activity of a Nostoc sp. PCC 7422 uptake hydrogenase-deficient mutant with high nitrogenase activity. AB - We describe a strategy to establish cyanobacterial strains with high levels of H(2) production that involves the identification of promising wild-type strains followed by optimization of the selected strains using genetic engineering. Nostoc sp. PCC 7422 was chosen from 12 other heterocystous strains, because it has the highest nitrogenase activity. We sequenced the uptake hydrogenase (Hup) gene cluster as well as the bidirectional hydrogenase gene cluster from the strain, and constructed a mutant (Delta hupL) by insertional disruption of the hupL gene. The Delta hupL mutant produced H(2) at 100 mumoles mg chlorophyll a ( 1) h(-1), a rate three times that of the wild-type. The Delta hupL cells could accumulate H(2) to about 29% (v/v) accompanied by O(2) evolution in 6 days, under a starting gas phase of Ar + 5% CO(2). The presence of 20% O(2) in the initial gas phase inhibited H(2) accumulation of the Delta hupL cells by less than 20% until day 7. PMID- 17131048 TI - Upper GI foreign body: an adult urban emergency hospital experience. AB - BACKGROUND: The efficacy of flexible endoscopy by a single endoscopist in the therapy of foreign body ingestion was assessed at an adult urban emergency hospital. METHODS: Fifty-one adult patients with upper GI foreign body ingestion treated at Detroit Receiving Hospital from 1988 to 2004 were identified. Endoscopic and hospital medical records were reviewed to evaluate etiology, treatment, and outcomes for these patients. RESULTS: The etiology was related to eating in 38(75%) patients, most of whom were eating meat; phytobezoars were seen in four, often after previous upper GI surgery. True foreign bodies were found in 13 patients (25%) and included a screwdriver, a ballpoint pen, spoons, coat hanger pieces, batteries, and latex gloves. Dysphagia was the most common symptom (75%); pain was common in patients with true foreign bodies, and 62% of this group had psychiatric difficulties or problems with drug abuse. Nearly 80% of the food-related group had post-surgical or other upper GI pathology. One patient had an esophageal stricture secondary to previous Sengstaken-Blakemore tube insertion. Flexible endoscopy was successful in extracting the foreign body in almost all (49) patients, with snare extraction the most common therapeutic modality. Both failures were of true foreign bodies that could not be safely removed. In one of these cases, it became necessary to employ the gallstone lithotripter, and the overtube was required in patients with metallic or sharp foreign bodies to protect the upper aerodigestive structures. CONCLUSIONS: Most upper GI foreign bodies are related to food impaction, with meat most often found. Underlying pathology is the rule and should be dealt with immediately. Flexible endoscopy is the treatment of choice for upper GI foreign body removal with near perfect success. PMID- 17131049 TI - Long-term outcome after surgical treatment of nonparasitic splenic cysts. AB - BACKGROUND: The optimal treatment for patients with nonparasitic splenic cysts is controversial. This study aimed to evaluate the clinical outcome of patients treated for a symptomatic splenic cyst, and to define a surgical strategy. METHODS: Spleen-preserving surgery (9 laparotomies and 6 laparoscopies) was performed for a primary cyst in six patients and a secondary cyst in nine patients. The median follow-up time was 37.5 months. Partial splenic resection was performed for eight patients and cyst decapsulation for seven patients. RESULTS: Cyst recurrence was observed in four patients after decapsulation of a primary splenic cyst, as compared with none after resection. Postoperative complications were encountered only after laparotomy (5/9). The median hospital stay was 3.5 days (range, 2-5 days) after laparoscopy, as compared with 9 days (range, 5-14 days) after laparotomy. CONCLUSIONS: Symptomatic splenic cysts should be treated laparoscopically. For patients with recurrent or suspected primary splenic cysts, laparoscopic partial splenectomy is preferable. For other cases, a laparoscopic decapsulation is advocated. PMID- 17131050 TI - Child and adolescent psychiatry training in Europe: differences and challenges in harmonization. AB - OBJECTIVE: To investigate the current situation of Child and Adolescent Psychiatry (CAP) training in European countries. In addition, current agenda of different organizations in order to harmonize the training across Europe are reported. METHOD: In order to collect data for this descriptive documentation on CAP training in European countries, we have communicated with "European Union of Medical Specialists Section on Child and Adolescent Psychiatry" (UEMS-CAP) and "The European Federation of Psychiatric Trainees" (EFPT) representatives of each country via e-mail. In addition, we used UEMS and EFPT annual forum minutes and web sites of national CAP societies to validate the data. RESULTS: The structure of CAP training has many differences between 34 European countries. For instance, in 32.4% of the countries, CAP is not a specialty in its own right but is mostly linked to general psychiatry. After medical school, the minimum training duration to become a CAP specialist ranges between 12 and 96 (mean: 59.71 +/- 17.1) months. While, a trainee should pass an examination to begin CAP training in 37.9% of the countries, 64.7% have an examination to graduate and both is the case in 29.7% of the countries. DISCUSSION: By the year 2006, European countries still have large differences in the structure of CAP training. It is assumed that the same holds true for content of training, which was not the focus of this documentation. UEMS-CAP, ESCAP (European Society of Child and Adolescent Psychiatry) and EFPT are the major bodies that have to manage the harmonization of CAP training across Europe. The obligatory conditions of a fruitful training, the high quality of teachers and teaching facilities, essential ingredients of an efficient training programme, are prone to be determined by evidence-based evaluations in the process of harmonization. PMID- 17131051 TI - Improving the thermal stability of lactate oxidase by directed evolution. AB - Lactate oxidase is used in biosensors to measure the concentration of lactate in the blood and other body fluids. Increasing the thermostability of lactate oxidase can significantly prolong the lifetime of these biosensors. We have previously obtained a variant of lactate oxidase from Aerococcus viridans with two mutations (E160G/V198I) that is significantly more thermostable than the wild type enzyme. Here we have attempted to further improve the thermostability of E160G/V198I lactate oxidase using directed evolution. We made a mutant lactate oxidase gene library by applying error-prone PCR and DNA shuffling, and screened for thermostable mutant lactate oxidase using a plate-based assay. After three rounds of screening we obtained a thermostable mutant lactate oxidase, which has six mutations (E160G/V198I/G36S/T103S/A232S/F277Y). The half-life of this lactate oxidase at 70 degrees C was about 2 times that of E160G/V198I and about 36 times that of the wild-type enzyme. The amino acid mutation process suggests that the combined neutral mutations are important in protein evolution. PMID- 17131052 TI - Evaluation of the anti-angiogenic effect of aloe-emodin. AB - The present study identified aloe-emodin (AE, a hydroxyanthraquinone from Aloe vera and other plants) as a new anti-angiogenic compound with inhibitory effects in an in vivo angiogenesis assay and evaluates its effects on specific key steps of the angiogenic process. AE inhibits endothelial cell proliferation, but this effect is not cell specific, since AE also inhibits tumor cell proliferation. Cell migration and invasion are not remarkably affected by AE. On the other hand, AE has different effects on endothelial and tumor cell gelatinases. Two main targets of the pharmacological action of AE as an anti-angiogenic compound seem to be urokinase secretion and tubule formation of endothelial cells. Finally, AE produces a remarkable photocytotoxic effect on tumor cells. Taken together, our data indicate that AE can behave both as an anti-tumor and an anti-angiogenic compound and suggest that AE could be a candidate drug for photodynamic therapy. PMID- 17131053 TI - Human progeroid syndromes, aging and cancer: new genetic and epigenetic insights into old questions. AB - Disorders in which individuals exhibit certain features of aging early in life are referred to as segmental progeroid syndromes. With the progress that has been made in understanding the etiologies of these conditions in the past decade, potential therapeutic options have begun to move from the realm of improbability to initial stages of testing. Among these syndromes, relevant advances have recently been made in Werner syndrome, one of several progeroid syndromes characterized by defective DNA helicases, and Hutchinson-Gilford progeria syndrome, which is characterized by aberrant processing of the nuclear envelope protein lamin A. Although best known for their causative roles in these illnesses, Werner protein and lamin A have also recently emerged as key players vulnerable to epigenetic changes that contribute to tumorigenesis and aging. These advances further demonstrate that understanding progeroid syndromes and introducing adequate treatments will not only prove beneficial to patients suffering from these dramatic diseases, but will also provide new mechanistic insights into cancer and normal aging processes. PMID- 17131054 TI - Protective effects of the PARP-1 inhibitor PJ34 in hypoxic-reoxygenated cardiomyoblasts. AB - To clarify the role of poly(ADP-ribose)polymerase-1 (PARP-1) in myocardial ischemia-reperfusion injury, we explored some effects of PJ34, a highly specific inhibitor of this enzyme, in hypoxic-reoxygenated (HR) H9c2 cardiomyoblasts. Compared to the control, HR cells showed signs of oxidative stress, marked PARP-1 activation, NAD(+) and ATP depletion and impaired mitochondrial activity. HR cardiomyoblasts were affected by both necrosis and apoptosis, the latter involving the nuclear translocation of apoptosis-inducing factor. In HR cardiomyoblasts treated with PJ34, oxidative stress and PARP-1 activity were decreased, and NAD(+) and ATP depletion, as well as mitochondrial impairment, were attenuated. Above all, PJ34 treatment improved the survival of HR cells; not only was necrosis significantly diminished, but apoptosis was also reduced and shifted from a caspase-independent to a caspase-dependent pathway. These results suggest that PARP-1 modulation by a selective inhibitor such as PJ34 may represent a promising approach to limit myocardial damage due to post-ischemic reperfusion. PMID- 17131055 TI - Matriptase and its putative role in cancer. AB - Tumor progression and metastasis are the pathologic effects of uncontrolled or deregulated invasive growth, a process in which proteases play a fundamental role. They mediate the degradation of extracellular matrix components and intercellular cohesive structures to allow migration of the cells into the extracellular environment and activate growth and angiogenic factors. In addition to metalloproteases and the plasminogen activation system, another protease, matriptase, contributes substantially to these processes. Matriptase is a type II transmembrane trypsin-like serine protease that is expressed by cells of epithelial origin and is overexpressed in a variety of human cancers. It has been suggested that this protease not only facilitates cellular invasiveness but may also activate oncogenic pathways. This review summarizes current knowledge about matriptase, its putative role in tumor initiation and progression, and its potential as a novel target in anti-cancer therapy. PMID- 17131056 TI - Directional mRNA transport in eukaryotes: lessons from yeast. AB - In eukaryotes, developmental processes and cell differentiation, as well as basic cellular functions require the propagation of information in an asymmetric manner. Localization of mRNA is a key mechanism to establish asymmetric cell fate. The first part of this review provides an overview of our current knowledge of motor protein-dependent mRNA transport in eukaryotes. The second part provides a more detailed description of the most comprehensively studied mRNA translocation complex to date: the ASH1 messenger ribonucleoprotein particle (mRNP) from Saccharomyces cerevisiae. During budding of yeast, the ASH1 mRNP transports cell fate determinants exclusively into the daughter cell. The core factors of the ASH1 mRNP have been identified, their interactions have been studied in detail, and the three-dimensional structure of its mRNA-binding protein, She2p, has been determined. Because no other mRNP has been studied in such detail, the ASH1 mRNP could serve as a model for asymmetric segregation of cell fate determinants in higher eukaryotes. PMID- 17131057 TI - Molecular characterization and expression of the antimicrobial peptide defensin from the housefly (Musca domestica). AB - A 430-bp cDNA encoding the insect antimicrobial peptide defensin was cloned from the housefly, and designated Musca domestica defensin (Mdde). The open reading frame of the cDNA encoded a 92-amino acid peptide with an N-terminal signal sequence followed by a propeptide that is processed by cleavage to a 40-amino acid mature peptide. Northern analysis and in situ hybridization identified the corresponding mRNA in the fat body of bacterially challenged houseflies and in the epidermis of the body wall of naive and challenged houseflies. The Gram negative bacterium (Escherichia coli) is a strong inducer of the gene. By RT-PCR, Mdde mRNA was also detected in naive and challenged insects. These findings suggest that the defensin gene is constitutively expressed in the epidermis of the housefly body wall. The predicted mature form of Mdde was expressed as a recombinant peptide in E. coli and Pichia pastoris. The recombinant Mdde expressed in Pichia was active against Gram-positive and some Gram-negative bacteria. PMID- 17131058 TI - Epigenetic regulation for the induction of meiosis. AB - The germ cell lineage is the sole cell lineage through which genomic information is transmitted into successive generations. To this end, germ cells undergo various specific differentiation steps including meiosis, whose regulation seems to correlate closely with fundamental mechanisms that create differences between germ cells and somatic cells. In mammals, meiosis is triggered by extra-embryonic stimuli such as retinoic acid, which may induce various intracellular molecular cascades promoting meiosis. In addition, the specific epigenetic status arranged in germ cells before and after induction of meiosis, including meiosis-specific transcription control based on histone methylation by a novel histone methyltransferase Meisetz, is also critical for its proper progression. PMID- 17131059 TI - Oncogenic mechanisms in myeloproliferative disorders. AB - Myeloproliferative disorders (MPDs) are clonal haematopoietic malignancies involving the abnormal proliferation of myeloid lineages. The World Health Organisation (WHO) classification of haematopoietic malignancies distinguishes MPDs from myelodysplastic/ myeloproliferative disorders and systemic mastocytosis. These malignancies frequently involve constitutive tyrosine kinase activity, resulting from either oncogenic fusion protein production or from point mutations. Chronic myelogenous leukaemia is the model used for studies of the consequences of such molecular defects. However, the heterogeneity of the clinical course of MPDs should be seen in a more rationale conceptual framework, including the many molecular events associated with these diseases. This review focuses on the various tyrosine kinase-related molecular mechanisms underlying both MPDs and rare diseases with myeloproliferative features. We pay particular attention to the newly identified JAK2 V617F mutation in polycythaemia vera, essential thrombocythaemia and idiopathic myelofibrosis and deal with disease heterogeneity and putative additional molecular mechanisms. PMID- 17131060 TI - The cold case: are rhinoviruses perfectly adapted pathogens? AB - Rhinoviruses, which cause common cold, belong to the Picornaviridae family, small non-enveloped viruses (diameter 15-30 nm) containing a single-stranded RNA genome (about 7 kb). Over 100 different rhinoviral serotypes have been identified thus far, establishing rhinoviruses as the most diverse group of Picornaviridae. Based on receptor binding properties, rhinoviruses are divided into two classes: the major group binding to intracellular adhesion molecule-1 and the minor group binding to the very low density lipoprotein receptors. Interactions between virus and the receptor molecules cause a conformational change in the capsid, which is a prerequisite for viral uptake. Rhinoviruses trigger a chemokine response upon infection that may lead to exacerbation of the symptoms of common cold, i.e. asthma and inflammation. The following review aims to summarize the knowledge about rhinoviral infections and discusses therapeutical approaches against this almost perfectly adapted pathogen. PMID- 17131061 TI - Recent structural studies of carbohydrate-binding modules. AB - Carbohydrate-binding modules (CBMs) are found in many carbohydrate-active enzymes. CBMs bind to a range of polysaccharides, their primary function being to increase the catalytic efficiency of the carbohydrate-active enzymes against soluble and/or insoluble substrates. CBMs bind to their target ligands with high specificities and affinities. Thus, CBM systems are excellent models to study the mechanism of protein-carbohydrate interaction. To date, CBMs have been classified into 45 different families and many structural and functional studies have been reported. At present, three-dimensional structures of CBMs from 31 different families have been determined. These structures demonstrate that the fold most commonly found in CBMs is the beta-sandwich. In the past few years, about 10 new structures from different families have been reported. These enable detailed classification of CBM structures. This article reviews recent structural and functional studies of CBMs and discusses the sub-classification of beta-sandwich CBMs. PMID- 17131062 TI - Antiquitin, a relatively unexplored member in the superfamily of aldehyde dehydrogenases with diversified physiological functions. AB - Antiquitin is a member of the aldehyde dehydrogenase superfamily. Sequence analyses indicate that the protein is highly conserved from plants to animals. The plant antiquitins are generally believed to play a role in osmoregulation and/or detoxification. The physiological functions of animal antiquitins remain largely elusive, their involvement in a number of human diseases has been implicated. PMID- 17131063 TI - Binding activity of norovirus and sapovirus to histo-blood group antigens. AB - Noroviruses (NoVs) and sapoviruses (SaVs) are causative agents of human gastroenteritis. There is increasing evidence that certain human NoV strains bind to histo-blood group antigens (HBGAs). We found that several NoV virus-like particles (VLPs) showed binding activity to HBGAs, while neither SaV genogroup I (GI) VLP nor SaV GV VLP showed such activity. PMID- 17131064 TI - Identification of genetic diversity of hepatitis E virus (HEV) and determination of the seroprevalence of HEV in eastern China. AB - Hepatitis E, caused by the hepatitis E virus (HEV), is endemic in China. However, the molecular characteristics of HEV circulating in eastern China and the seroprevalence of HEV infection in eastern China are relatively unknown. In this study, 25 HEV strains, isolated from sporadic hepatitis E cases in eastern China, were sequenced in the RNA-dependent RNA polymerase region and subjected to phylogenetic analysis. These HEV strains were 74.6-98.7% identical in nucleotides and were all clustered into HEV genotype 4. Most of them formed new sub-genotypes and revealed a high degree of genetic variance. In addition, 12,052 serum samples were collected from people of different ages, living in urban or rural areas in eastern China. Anti-HEV IgG activity was detected in 2073 (17.20%). The prevalence of anti-HEV IgG significantly increased with age (P<0.0001), ranging from 7.92% in children (<10 years old) to 21.48% among older persons (>or=60 years old). Moreover, statistical analysis showed that there was a significant difference between rural and urban areas, with higher prevalence for people living in rural neighborhoods (P<0.001). PMID- 17131065 TI - Molecular epidemiology and phylogenetic analysis of a marine fish infectious spleen and kidney necrosis virus-like (ISKNV-like) virus. AB - Infectious spleen and kidney necrosis virus-like (ISKNV-like) virus causes a serious systemic disease with high morbidity and mortality of freshwater and marine fishes. Based on the ISKNV putative major capsid protein (MCP), the vascular endothelial growth factor (VEGF), the mRNA capping enzyme (Capping), and the tumor necrosis factor receptor-associated protein (TNFR) genes, primers were designed and used in PCR to determine the host range of ISKNV-like viruses. From the sampling of >1600 marine fishes representing 6 orders, 25 families, and 86 species collected in the South China Sea, 13 cultured fish species (141 fish) and 39 wild fish species (102 fish) were confirmed hosts of ISKNV-like viruses. The average percentage of infection of ISKNV-like viruses was 14.6%. The results from phylogenetic analysis of these genes revealed that ISKNV-like viruses could be placed into two clusters: cluster I was more related to ISKNV; cluster II included OSGIV (orange-spotted grouper iridovirus) and RBIV (rock bream iridovirus), and was quite different from ISKNV. The results of this study can contribute to the prediction and prevention of ISKNV disease outbreaks. PMID- 17131066 TI - Lung adenocarcinoma metastasizing to cerebellopontine angle schwannoma (collision tumor). AB - Collision tumors are composed of two histologically distinct neoplasms which present at one location. Collision of a malignant tumor originating outside the central nervous system with a primary intracranial neoplasm is a rare event. Here we report on a CPA schwannoma containing a metastasis of adenocarcinoma in a 57 years old woman. Searching for the primary tumor resulted in the detection of a carcinoma in the superior lobe of the left lung. PMID- 17131067 TI - Chiari I malformation associated with atlanto-axial dislocation: focussing on the anterior cervico-medullary compression. AB - BACKGROUND: Chiari I malformation with atlantoaxial dislocation may cause both posterior and anterior cervicomedullary compression. We studied the clinicoradiological features and surgical outcome in patients having Chiari I malformation with atlantoaxial dislocation. METHOD: Thirty-nine patients with Chiari I malformation with atlanto-axial dislocation underwent preoperative and follow-up neurological status assessment. In Chiari I malformation with reducible atlanto-axial dislocation (n = 11), a direct posterior stabilization was done. In Chiari I malformation with irreducible atlanto-axial dislocation (n = 28), a single stage transoral decompression with posterior stabilization and/or posterior decompression and duraplasty were done in 18 patients. In 10 patients, only posterior decompression and/or posterior stabilization was performed. Seven among the latter patients subsequently deteriorated and required transoral decompression. Comparison of mean neurological status scores of patients with Chiari I malformation with irreducible atlanto-axial dislocation who underwent single stage transoral decompression with posterior stabilization versus the posterior procedure alone was done using T-test and proportional significance also calculated. FINDINGS: Patients with Chiari I malformation with atlanto-axial dislocation have a high incidence of long tract signs and sphincteric disturbances with a decrease in the mean foramen magnum diameter. The mean neurological status scores of patients with Chiari I malformation with irreducible atlanto-axial dislocation who underwent single stage transoral decompression with posterior stabilization were significantly better than those patients who underwent the posterior procedure alone. The latter patients also showed significant clinical improvement following transoral decompression. In the presence of Chiari I malformation with reducible atlanto-axial dislocation, reduction and stabilization of atlanto-axial dislocation resulted in neurological improvement. The follow up neurological status scores of these patients improved after surgical intervention even in the presence of poor preoperative grades. CONCLUSIONS: Patients with Chiari I malformation should be investigated for the presence of atlanto-axial dislocation. In case atlantoaxial dislocation coexists, priority must be given to relieving anterior cervicomedullary compression. PMID- 17131068 TI - Neurosurgical management of inadequately embolized intracranial aneurysms: a series of 17 consecutive cases. AB - OBJECTIVE: Inadequately embolized aneurysms (IEA) are coiled aneurysms with a significant remnant (>5%), initially or after recanalisation, or with a coil extrusion deemed too thrombogenic or threatening the blood flow in the parent vessel. Our objective is to report our experience with the surgical clipping (SC) of a consecutive series of 17 IEA considered as not appropriate for an additional endovascular procedure. METHODS: Between February 1996 and April 2006, we evaluated 523 ICA in 380 patients of whom 192 underwent coil embolisation (CE), 117 with complete occlusion (61%), 47 with near complete occlusion (> or = 95%), 9 with partial occlusion (<95%), and 19 without any coil delivery (attempted embolisation). Of the 173 ICA embolized one or two times, at their radiological follow-up 15 (8.6%) were considered as IEA and not appropriate for an additional CE. Two IEA treated endovascularly before February 1996 were added to this series. The female/male ratio was 0.47 with an average age of 54 years (range, 37 65). All cases were located on the anterior circulation except the last one. The 17 IEA were treated by SC either because of an aneurysm remnant deemed not accessible to a further CE and large enough for direct clipping or because the risks of a thromboembolic event related to extruded coils was too high. RESULTS: SC was complete in all 17 cases, confirmed angiographically. Postoperatively, the clinical status of two patients deteriorated slightly but transiently. Our surgical experience with this series led us to classify IEA into five types, in three groups: group A (with one type: type A) was the most important group (n = 11) with IEA characterized by an aneurysm residue allowing direct SC, as assessed preoperatively; group B (n = 4) comprised aneurysms with a residue smaller than predicted and showing parent vessel stenosis when a clip was applied to the neck residue requiring the fundus full of coils to be removed followed by either clip application to the neck residue (type B1, n = 3) or suture if the remnant was too small (type B2, n = 1); and group C (n = 2) grouping cases requiring coil extraction through the parent vessel (type C2, n = 1) or through the fundus (type C2, n = 1). CONCLUSIONS: With this series of IEA, we observed that open surgery of type A and C aneurysms can be a straightforward procedure. Our experience with type B IEA encourages us to wait for a sufficient aneurysm residue before performing SC because of the potential difficulties that may be encountered by the surgeon, particularly in type B2. SC of IEA was very effective with complete occlusion and no permanent morbidity in all 17 cases. PMID- 17131069 TI - Intra-operative diagnosis and image-guided management of an intracerebral haemorrhage occurring during ultrasound-guided biopsy. AB - Haemorrhagic complications occurring after burr-hole procedures are diagnosed only in symptomatic patients or when postoperative imaging is performed routinely. We report the development of an intracerebral haematoma which occurred during ultrasound-guided burr-hole biopsy. Real-time ultrasound through the same burr-hole enabled us to determine the dynamics of the bleeding and its terminal volume. The operation was finished without further complications and the patient did not experience an impairment of her neurological state. Intra-operative ultrasound is capable of detecting "invisible" complications during burr-hole procedures. PMID- 17131070 TI - The single-staged approach to the surgical management of abdominal wall hernias in contaminated fields. AB - INTRODUCTION: The surgical treatment of large ventral hernias with accompanying contamination is challenging. We have reviewed our institution's experience with single-staged repair of complex ventral hernias in the setting of contamination. METHODS: We retrospectively reviewed the medical records of all patients who underwent ventral hernia repairs in the setting of a contaminated field. Pertinent details included baseline demographics, reason for contamination, operative technique and details, postoperative morbidity, mortality and recurrence rates. RESULTS: Between December 1999 and January 2006, 19 patients were identified with ventral hernia repairs performed in contaminated fields. There were 6 males and 13 females with a mean age of 61 years (40-82), ASA 3.2 (2 4), and BMI of 34 kg/m(2) (20-65). Fourteen patients had prior mesh: prolene (9), composix (3), goretex (1), and alloderm (1). Reasons for contamination included: mesh infection (14), enterocutaneous fistula (7), concomitant bowel resection (8), chronic non-healing wound (2), and necrotizing fasciitis (1). Operative approaches included primary repair (3), component separation without reinforcement (2), and with prosthetic reinforcement (9). In five patients the fascia could not be reapproximated in the midline and the defect was bridged with surgisis (1), Marlex (1), lightweight polypropylene (1) placed in the retrorectus space, and alloderm (2). Mean operative time was 260 min (90-600). Twelve postoperative complications occurred in nine (47%) patients and included wound infection (6), respiratory failure (1), ileus (2), postoperative hemorrhage (1), renal failure (1), and atrial fibrillation (1). One patient died in this series. During routine follow-up two recurrences were identified by physical exam. CONCLUSIONS: This study shows that single-stage treatment of ventral hernias in contaminated fields can be accomplished with a low recurrence rate and acceptable morbidity in these extremely challenging patients. PMID- 17131071 TI - Surgical technique and complications during laparoscopic repair of diaphragmatic hernias. AB - Diaphragmatic hernias can present as retrocostoxiphoid hernias (RCXH) or diaphragmatic dome hernias. The RCXH include the Larrey hernia (LH), the Morgagni hernia (MH), and the Larrey-Morgagni hernia (LMH). These congenital hernias are usually asymptomatic, and the diagnosis is simplified by two exams: chest X-ray, and thoraco-abdominal computed tomography (CT) scan. The potential risk in this condition is small-bowel incarceration in the hernia defect and subsequent obstruction. We report two cases of LH and one case of LMH treated by laparoscopy between February 2004 and October 2005, with a review of the surgical techniques. Two different laparoscopic techniques were used: the tension-free technique, and resection of the hernia sac with closure of the defect and reinforcement by prosthesis. One patient presented a postoperative cardiac tamponade due to a clip induced bleeding of an epicardial artery at the inferior surface of the heart. Treatment by laparoscopy is feasible, but a consensus regarding the best laparoscopic repair is needed. PMID- 17131072 TI - The comparison of laparoscopic and open ventral hernia repairs: a prospective randomized study. AB - BACKGROUND: The laparoscopic approach has emerged in the search for a surgical technique to decrease the morbidity associated with conventional repair of ventral hernias. In this study we aimed to compare the results of our open and laparoscopic ventral hernia repairs prospectively. METHODS: Between January 2001 and October 2005, a total of 46 patients diagnosed with ventral hernias (primary and incisional) who were admitted to our surgical unit and accepted to be included in this study group were examined. All patients were divided into laparoscopic repair (n = 23) and open repair (n = 23) subgroups in a randomized fashion. The patients' demographic characteristics, operation times, body mass indices, sizes of fascial defects, hernia locations, durations of hospital stay, presence and degrees of postoperative pain, and postoperative minor and major complications were analysed and compared. All the data were expressed as means +/ SDs. Chi-square and Wilcoxon tests were used for statistical analysis, and P < 0.05 was accepted as a significant statistical value (SPSS 11.0 for Windows). RESULTS: The demographic characteristics of both groups were similar. Women predominated, especially in the laparoscopy group (P < 0.05). The comparison of the results revealed that the major advantage of laparoscopy was the shortened postoperative hospital stay and the reduced incidence of mesh infection (P < 0.05, P < 0.05). On the other hand, operation time was significantly longer in the laparoscopy group (P < 0.05). The major complications encountered in the laparoscopy group were ileus and a missed enterotomy. The most frequent minor complication was seroma, which was significantly more frequent in the laparoscopy group (P < 0.05). Postoperative pain assessment revealed similar results in both groups (P > 0.05). CONCLUSIONS: The laparoscopic approach appears to be as effective as open repairs in the treatment of ventral hernias. Advanced surgical skill, laparoscopic experience and high technology are mandatory factors for successful ventral hernia repair. PMID- 17131073 TI - Increased prevalence of concentric left ventricular hypertrophy in African Americans: will an epidemic of heart failure follow? AB - Left ventricular hypertrophy (LVH), used in this review to denote abnormally increased left ventricular (LV) mass, is an important cardiac trait because of its association with numerous adverse cardiovascular outcomes including myocardial infarction and heart failure. LV mass is typically assessed by noninvasive cardiac imaging (echocardiography or MRI); electrocardiography is an insensitive measure. There are two predominant types of hypertrophy: concentric, where LV wall thickness is increased relative to cavity dimensions, and eccentric, where LV wall thickness is not increased relative to cavity dimensions. Several large studies indicate that the prevalence of concentric LVH is higher in African-Americans versus whites. Although there are data to suggest that concentric LVH results in systolic heart failure in animal models, such data are lacking in humans. How concentric LVH affects the prevalence of systolic and diastolic heart failure in African-Americans needs further study. Given the large burden of LVH among African-Americans, such data are needed to estimate the expected burden and type of heart failure which will occur in the future in this population. PMID- 17131074 TI - Atherosclerosis imaging and heart failure. AB - Coronary atherosclerosis is the most important primary etiologic factor predisposing to the development of heart failure. The mechanisms by which coronary atherosclerosis lead to heart failure likely involve the initial development of regional myocardial dysfunction, later progressing to global ventricular failure and symptomatic congestive disease. A variety of imaging strategies have been investigated for their value in identifying and characterizing markers of atherosclerosis in the effort to detect early cardiac disease. Non-invasive imaging techniques for assessing anatomic or functional manifestations of atherosclerosis include carotid ultrasonography, coronary computed tomography, cardiovascular magnetic resonance imaging, brachial artery reactivity testing, and the ankle-brachial index. Many of these imaging methods are shown to have accuracy, reliability, and the potential to add value to an office-based cardiovascular risk assessment. Further development of such imaging methods could facilitate early intervention in the development of myocardial dysfunction while enhancing our understanding of the natural course of atherosclerotic disease. PMID- 17131075 TI - Imaging of myocardial dyssynchrony in congestive heart failure. AB - Heart failure constitutes a major health problem in USA and Europe. Angiotensin converting enzyme inhibitors and _ blockers were shown to reduce morbidity and mortality in patients with CHF. Yet, their effectiveness is limited. A significant number of patients with heart failure manifest myocardial conduction abnormalities. Conduction abnormalities, especially in the form of left bundle branch block (LBBB) may be associated with abnormal mechanical function. Several studies demonstrated that these patients may gain benefit from biventricular (BiV) pacing in terms of improvement in exercise tolerance, heart failure morbidity and even decreased mortality. BiV pacing was also associated with improvement in ejection fraction, reduction in the extent of mitral regurgitation and a decrease in cardiac size (reverse remodeling). However, a significant number of patients do not gain benefit from biventricular pacing despite having conduction abnormalities. The underlying reason is that the electrical activity may not closely reflect mechanical activity. Several imaging modalities and techniques have been proposed to improve the selection of patients who may benefit from biventricular pacemakers. Of those, echo-Doppler, and especially, Tissue Doppler Imaging has been demonstrated as important tools for evaluating patients for cardiac resynchronization therapy (CRT) and following their response. The advantages of echo include accessibility, portability, its cost and a high temporal resolution. Yet, it is limited by its acoustic windows and scanning angles. MRI is a useful tool for evaluating patients for CRT by providing 3-D image of myocardial function. However, it is limited for follow-up after implantation due to its cost and a potential damage to the patients or pacemakers. Dyssnchrony imaging is a rapidly evolving field. New imaging techniques such as speckle tracking are promising and close update is needed to keep track of the developments and the changes in this exciting field. PMID- 17131076 TI - Imaging microvascular obstruction and its clinical significance following acute myocardial infarction. AB - Obstruction of the coronary microvasculature contributes to the pathophysiology of MI and adversely affects post-MI recovery. This "no-reflow" phenomenon resulting from microvascular obstruction is an indicator of lack of adequate tissue perfusion within the infarcted myocardium, even after restoration of epicardial blood flow. Regions of microvascular obstruction can be detected and quantifed because of rapid advances in and refinement of imaging technologies over the past decade. This article focuses on the non-invasive imaging modalities used to assess MO, discusses the prognostic implications of MO, and briefly addresses strategies for reducing MO. PMID- 17131077 TI - Magnetic resonance imaging in the evaluation of non-ischemic cardiomyopathies: current applications and future perspectives. AB - Patients with non-ischemic cardiomyopathy often represent a diagnostic challenge, and correct etiologic diagnosis may influence outcomes. Lately, delayed myocardial enhancement MR imaging has been developed and is currently being used for a growing number of clinical applications. On delayed enhancement MR images, scarring or fibrosis appears as an area of high signal intensity, and the pattern by which this enhancement occurs in the myocardium allows distinction of many different pathologies. In nonischemic cardiomyopathy, the delayed enhancement usually does not occur in a coronary artery distribution and is often midwall rather than subendocardial or transmural. It could also guide myocardial biopsy to an affected area, increasing its yield. Cardiac magnetic resonance imaging has now a definitive role in clinical practice, and its capability to non-invasively provide high resolution images of the heart with good tissue characterization is redefining the understanding of the conditions that can adversely affect the myocardium. PMID- 17131078 TI - Nuclear imaging in cardiac cell therapy. AB - Cardiac stem cell therapy is an innovative and promising therapeutic approach for heart failure. However, despite an increasing body of existing experimental and human data, it still presents a substantial challenge for basic scientists and clinical researchers. Several issues concerning biologic mechanisms of therapy remain to be answered, and unequivocal proof of clinical efficacy is needed. The variety of different available cell types and different methods for cell delivery to the myocardium raises further questions about the most useful therapeutic approach. Nuclear imaging not only provides accurate noninvasive information about myocardial perfusion, contractile function and viability, which enables assessment of clinical benefits of therapy. The rapidly developing field of molecular imaging has also brought up more specific tracers targeting cellular and subcellular biologic events, which are expected to shed more light upon mechanisms of cell therapy. Moreover, nuclear imaging is well suited for tracking of transplanted cells by use of direct radionuclide labeling or genetic labeling with reporter genes that can be targeted by radioactive reporter probes. Such a broad spectrum of available in vivo information is expected to significantly impact the future development of cell therapy towards a clinically accepted treatment. PMID- 17131080 TI - Imidacloprid residues in fruits, vegetables and water samples from Palestine. AB - The aim of this work was to report on imidacloprid [IUPAC name 1-(6-chloro-3 pyridylmethyl)-N-nitroimidazolidin-2-ylideneamine] residues in some vegetables, fruits, and water samples collected from the West Bank, Palestine, in 1998 and 1999. Imidacloprid and its derivatives in the study samples were extracted by methanol/water and oxidized into 6-chloronicotinic acid and subsequently derivatized into 6-chloronicotinic acid trimethylsilyl ester before being determined by GC/MS. Imidacloprid residues were detected in more than half of the analyzed samples. The highest and lowest imidacloprid concentrations were found in eggplant (0.46 mg/kg) and green beans (0.08 mg/kg), respectively. An increase of 11-120% in imidacloprid concentration in the 1999 samples was observed when compared with those of 1998. This may suggest imidacloprid accumulation in the soil and/or increased use by local farmers. The imidacloprid residue concentrations in several crops were found to exceed the CODEX maximum residue limit. PMID- 17131079 TI - Novel and potential future biomarkers for assessment of the severity and prognosis of chronic heart failure : a clinical review. AB - Over the last two decades, the pathophysiology and biomolecular basis of heart failure syndrome has reached sound and more comprehensive understanding. This knowledge has allowed expert researchers and clinicians to explore an entirely new spectrum of potential biochemical markers derived from different cellular and signaling pathways that lead to myocardial hypertrophy, chronic damage of the myocyte, apoptosis, and, ultimately, myocardial remodeling. Indeed, the link between myocardial remodeling and adverse outcomes, as well as the recognition of the myocardial interstitium as a multifunctional dynamic entity strongly influenced by systemic neurohormonal and inflammatory activation, has provided a solid ground for research of biomarkers that might correlate with severity and prognostication in chronic heart failure. This paper reviews and summarize recent literature on some of the most interesting circulating biomarkers with potential use for the stratification of patients with chronic heart failure. PMID- 17131081 TI - Using lichen chemistry to assess airborne tungsten and cobalt in Fallon, Nevada. AB - This paper describes the use of lichen chemistry to assess airborne tungsten and cobalt in Fallon, Nevada, where a cluster of childhood leukemia has been on going since 1997. Lichens and their rock substrates were collected from Rattlesnake Hill within Fallon as well as from four different rock outcrops located north, east, south, and west of Fallon and at least 20 km away from the town center. In the lichens themselves, W and Co are significantly higher within Fallon than in the combined control site outside of Fallon. In the rock substrates of the lichens, no differences exist in W and Co. The W and Co differences in lichens cannot be attributed to substrate geochemistry. Fallon is distinctive in west central Nevada for high airborne W and Co, and given its cluster of childhood leukemia, it stands to reason that additional biomedical research is in order to test directly the leukogenicity of combined airborne W and Co. PMID- 17131082 TI - Hydrodynamic simulation of river Yamuna for riverbed assessment: a case study of Delhi region. AB - A well known river hydrodynamic model RiverCAD has been used to simulate and visualize flood scenarios for different designated flood flows under complex riverbed geometry with several man made structures like bridges and barrages. The model applied successfully for the stretch of 23 km in the Yamuna floodplain of Delhi region from Wazirabad barrage in the upstream to Okhla barrage. Flood flows for various return periods namely once in 10, 25, 50 and 100 years were estimated based on recorded flow data for the period of 1963 to 2003 using standard flood frequency analysis techniques. The simulation results were compared and the model was calibrated with water surface elevation records of the previous floods at various barrage and bridge locations. Simulation results enabled prediction of maximum water levels, submergence scenarios and land availability under different designated flood flows for riverbed assessment, development and management. PMID- 17131083 TI - Deposition in boreal forests in relation to type, size, number and placement of collectors. AB - Open precipitation and throughfall was collected at a Norway spruce stand in Finland using funnel-type collectors and at a black spruce stand in Canada using trough-type collectors. The presence or absence of a rim on the funnel, funnel diameter (9, 14 and 20 cm) and length of sampling period (1, 2 and 4 weeks) on monthly values were evaluated at the Norway spruce stand, and the number of collectors required for defined levels of accuracy and precision of throughfall loads to be reached and the influence of the spatial arrangement of collectors on solute concentrations was studied at both stands. The presence of a rim had no significant effect on open precipitation and throughfall amounts, but did on throughfall DOC, Ca(2+), Mg(2+), K(+), Na(+) and Cl(-) ion loads. Deposition loads increased with decreasing funnel diameter; for open precipitation, this was due to increased catch efficiency while for throughfall the increase was attributed to canopy interaction and leaching of litter trapped in the collectors. Calculated monthly H(+) loads decreased and those for all other constituents increased with collection period length. Using 15 collectors at the Norway spruce stand would allow throughfall loads to be determined to within 20% of the true mean weekly value with a confidence level of 95% for most solute, but not for NH(4) (+)-N, NO(3) (-)-N, Mg(2+) and SO(4) (2-)-S. Using 15 trough collectors, the same confidence level at the more heterogeneous black spruce stand would only be achieved for H(+), Cl(-), DOC and SO(4) (2-)-S loads. In both stands, using either random or systematic placements of throughfall collectors gave similar results. PMID- 17131084 TI - Measurement of respiratory rate from the photoplethysmogram in chest clinic patients. AB - OBJECTIVE: We studied the application of our algorithm for the robust extraction of respiratory information from the pulse oximeter signal acquired from a selection of patients attending the chest clinic. METHODS: Photoplethysmograms were obtained from 16 individuals: 13 patients with various conditions in the respiratory ward and three healthy subjects. Wavelet transforms were generated from which respiratory information was extracted to obtain a measure of respiratory rate. This measured rate was compared with the respiratory rate determined by one of a variety of other means (a digital end tidal CO(2) signal, the output from a non-invasive ventilation device, or a switch actuated by the patient or observer.) RESULTS: Respiratory rates varied from 6.2 to 35.8 breaths per minute (bpm). The oximeter rate determined through our method matched the marker rate obtained for all patients to within 1 bpm. CONCLUSION: The technique allows the measurement of respiratory rate directly from the photoplethysmogram of a pulse oximeter, and leads the way for development of a simple non-invasive combined respiration and saturation monitor useful for patients with all forms of breathlessness. PMID- 17131085 TI - Asymmetric spatiotemporal evolution of prebiotic homochirality. AB - The role of asymmetry on the evolution of prebiotic homochirality is investigated in the context of autocatalytic polymerization reaction networks. A model featuring enantiometric cross-inhibition and chiral bias is used to study the diffusion equations controlling the spatiotemporal development of left and right handed domains. Bounds on the chiral bias are obtained based on present-day constraints on the emergence of life on early Earth. The viability of biasing mechanisms such as weak neutral currents and circularly polarized UV light is discussed. The results can be applied to any hypothetical planetary platform. PMID- 17131087 TI - Polyynes and cyanopolyynes: their synthesis with the carbon arc gives the same abundances occurring in carbon-rich stars. AB - Carbon vapour generated from a carbon arc or by laser ablation of graphite is reactive with simple molecules and atoms producing end-capped polyyne chains. With these techniques both hydrogen-terminated polyynes as well as monocyano- and dicyanopolyynes have been produced. Experiments based on arcing graphite electrodes can reproduce the molecular distribution of polyynes existing around carbon-rich AGB stars. In fact, it has been found that the relative abundances of the polyynes produced in carbon arc in vacuum decreases by a factor between 3 and 5 as the chain length increases by a C(2) unit. An analogous trend has been observed both for polyynes and cyanopolyynes in the circumstellar environment around carbon-rich stars. This fact suggests that the mechanism of formation of the polyynes in the carbon arc may be similar to that occurring in the surroundings of the carbon-rich stars. Polyynes and cyanopolyynes represent authentic prebiotic molecules which appear quite ubiquitous in the cosmos and should have played a role in the early organic chemistry preceding the appearance of life. PMID- 17131088 TI - Gas-phase reactions in extraterrestrial environments: laboratory investigations by crossed molecular beams. AB - We have investigated gas-phase reactions of N((2)D) with the most abundant hydrocarbons in the atmosphere of Titan by the crossed molecular beam technique. In all cases, molecular products containing a novel CN bond are formed, thus suggesting possible routes of formation of gas-phase nitriles in the atmosphere of Titan and primordial Earth. The same approach has been recently extended to the study of radical-radical reactions, such as the reaction of atomic oxygen with the CH(3) and C(3)H(5) radicals. Products other than those already considered in the modeling of planetary atmospheres and interstellar medium have been identified. PMID- 17131089 TI - Self-inflicted fear of evolution. AB - Commonly calculated zero probabilities for synthesis of a given protein sequence by chance are that small because the sizes of the proteins taken for the calculations are too large (over 100 residues). Same estimate for 20-30 residue chains makes the chance close to 1. PMID- 17131090 TI - Fresh new air in space? 'MoMa' on the A.S.I. launch pad. AB - The present project has been developed because of the desire to unify the research lines in the A.S.I. 'Medicine & Biotechnology' area into one research line that could satisfy the interests of all of the collaborative groups and at the same time could pursue a relevant social goal. A 6 month feasibility study (SF) called MoMa was carried out in the ASI framework. During the SF the know-how and the tools already available in the national scientific community have been assessed, selected and evaluated even with the important contribution of Small and Medium-size Enterprises (SME) and of Italian industries already involved in Space Research. As result of the SF MoMa, all of the participants decided to combine all the efforts together and define, with all the know-how and the available technologies, one strategic topic, the "Aging" with a special attention to the Quality of Life (QoL). The space environment is a unique laboratory to study the reaction of living organisms (especially humans) to microgravity and cosmic radiation. The study of the effects of these two variables at the molecular and cellular levels will shed light on the response of cells and living organisms to adverse stimulations that are always present even on Earth and will help us able to develop the best strategies to protect the organisms from the progressive structural and functional decline related to Aging. Relevant spin offs on Earth and also relevant industrial applications are the expected outputs of this project. PMID- 17131091 TI - Mars primordial crust: unique sites for investigating proto-biologic properties. AB - The Martian meteorite collection suggests that intact outcrops or boulder-scale fragments of the 4.5 Ga Martian crust exist within tens of meters of the present day surface of Mars. Mars may be the only planet where such primordial crust samples, representing the first 100 Ma of a planet's environment, are available. The primordial crust has been destroyed on Earth by plate tectonics and other geological phenomena and is buried on the Moon under hundreds or thousands of meters of megaregoltih. Early Mars appears to have been remarkably similar to early Earth, and samples of rock from the first few Ma or first 100 Ma may reveal "missing link" proto-biological forms that could shed light on the transition from abiotic organic chemistry to living cells. Such organic snapshots of nascent life are unlikely to be found on Earth. PMID- 17131092 TI - From Never Born Proteins to Minimal Living Cells: two projects in synthetic biology. AB - The Never Born Proteins (NBPs) and the Minimal Cell projects are two currently developed research lines belonging to the field of synthetic biology. The first deals with the investigation of structural and functional properties of de novo proteins with random sequences, selected and isolated using phage display methods. The minimal cell is the simplest cellular construct which displays living properties, such as self-maintenance, self-reproduction and evolvability. The semi-synthetic approach to minimal cells involves the use of extant genes and proteins in order to build a supramolecular construct based on lipid vesicles. Results and outlooks on these two research lines are shortly discussed, mainly focusing on their relevance to the origin of life studies. PMID- 17131093 TI - Gamma-radiation induced polymerization of a chiral monomer: a new way to produce chiral amplification. AB - The treatment of the terpene beta(-)pinene with gamma radiation (at dose level: 150, 300 and 600 kGy) causes its polymerization into a resin and into a dimer. The yield of the resin and of the dimer appears to be linearly dependent to the radiation dose. The structure of the products was studied by FT-IR spectroscopy also in comparison to a reference beta(-)pinene resin prepared by cationic polymerization. A highly ordered structure was found in the case of the radiopolymer in comparison to the resin from cationic polymerization. Polarimetric measurements have shown astonishing enhancement in the optical activity of the radiopolymer and radiodimer in comparison to the starting optical activity of the beta(-)pinene monomer. Also DSC (differential scanning calorimetry) data supports the unexpected highly ordered structure for the beta( )pinene radiopolymer in comparison to the resin prepared by cationic polymerization. The results have been discussed in terms of amplification of chirality caused by gamma radiation and the implications of this fact on the mechanism of chiral amplification on prebiotic molecules. PMID- 17131094 TI - Damage to the oxygen-evolving complex by superoxide anion, hydrogen peroxide, and hydroxyl radical in photoinhibition of photosystem II. AB - Under strong illumination of a photosystem II (PSII) membrane, endogenous superoxide anion, hydrogen peroxide, and hydroxyl radical were successively produced. These compounds then cooperatively resulted in a release of manganese from the oxygen-evolving complex (OEC) and an inhibition of oxygen evolution activity. The OEC inactivation was initiated by an acceptor-side generated superoxide anion, and hydrogen peroxide was most probably responsible for the transportation of reactive oxygen species (ROS) across the PSII membrane from the acceptor-side to the donor-side. Besides ROS being generated in the acceptor-side induced manganese loss; there may also be a ROS-independent manganese loss in the OEC of PSII. Both superoxide anion and hydroxyl radical located inside the PSII membrane were directly identified by a spin trapping-electron spin resonance (ESR) method in combination with a lipophilic spin trap, 5-(diethoxyphosphoryl)-5 phenethyl-1-pyrroline N-oxide (DEPPEPO). The endogenous hydrogen peroxide production was examined by oxidation of thiobenzamide. PMID- 17131095 TI - C3 photosynthesis in silico. AB - A computer model comprising light reactions, electron-proton transport, enzymatic reactions, and regulatory functions of C3 photosynthesis has been developed as a system of differential budget equations for intermediate compounds. The emphasis is on electron transport through PSII and PSI and on the modeling of Chl fluorescence and 810 nm absorptance signals. Non-photochemical quenching of PSII excitation is controlled by lumenal pH. Alternative electron transport is modeled as the Mehler type O2 reduction plus the malate-oxaloacetate shuttle based on the chloroplast malate dehydrogenase. Carbon reduction enzymes are redox-controlled by the ferredoxin-thioredoxin system, sucrose synthesis is controlled by the fructose 2,6-bisphosphate inhibition of cytosolic FBPase, and starch synthesis is controlled by ADP-glucose pyrophosphorylase. Photorespiratory glycolate pathway is included in an integrated way, sufficient to reproduce steady-state rates of photorespiration. Rate-equations are designed on principles of multisubstrate multiproduct enzyme kinetics. The parameters of the model were adopted from literature or were estimated from fitting the photosynthetic rate and pool sizes to experimental data. The model provided good simulations for steady-state photosynthesis, Chl fluorescence, and 810 nm transmittance signals under varying light, CO2 and O2 concentrations, as well as for the transients of post illumination CO2 uptake, Chl fluorescence induction and the 810 nm signal. The modeling shows that the present understanding of photosynthesis incorporated in the model is basically correct, but still insufficient to reproduce the dark light induction of photosynthesis, the time kinetics of non-photochemical quenching, 'photosynthetic control' of plastoquinone oxidation, cyclic electron flow around PSI, oscillations in photosynthesis. The model may find application for predicting the results of gene transformations, the analysis of kinetic experimental data, the training of students. PMID- 17131096 TI - Effect of methanol intoxication on specific immune functions of albino rats. AB - Our previous studies revealed that methanol intoxication significantly altered the non-specific immune functions in albino rats. The present investigation focuses on the effect of methanol on certain specific immune functions of cell mediated immunity such as footpad thickness, leukocyte migration inhibition test (LMI) and antibody levels. In addition, serum interleukins (IL-2, IL-4, TNF-alpha and IFN-gamma), and splenic lymphocyte subsets were measured after an immune challenge. The specific immune function tests were carried out in three different groups of albino rats, which include control, 15 and 30 days methanol intoxication. Our study reports that animal body weight, organ weight ratio, lymphoid cell counts, footpad thickness, antibody titer, IL-2, TNF-alpha, IFN gamma, Pan T cell, CD4, macrophages, MHC class II molecule expression, and B cell counts were significantly decreased compared to control animals nevertheless, LMI, IL-4, and DNA single strand breakage were increased significantly. Plasma corticosterone level was significantly increased in the 15 days group whereas the 30 days methanol intoxication group showed considerable decrease in corticosterone level compared with control animals. Therefore, our investigation concluded that repeated exposure of methanol profoundly suppressed the cell mediated and humoral immune functions in albino rats. PMID- 17131097 TI - DMPS reverts morphologic and mitochondrial damage in OK cells exposed to toxic concentrations of HgCl2. AB - Mercuric chloride (HgCl(2)) is a highly toxic compound, which can cause nephrotoxic damage. In the present study effects of HgCl(2) on mitochondria integrity and energy metabolism, as well as antidotal effects of 2,3 dimercaptopropane-1-sulfonate (DMPS) were investigated in the opossum kidney derived cell line (OK). OK cell monolayers were incubated during 0, 1, 3, 6, and 9 h in serum-free culture medium containing 15 microM HgCl(2), either in the absence or in the presence of 60 microM DMPS in a 1:4 ratio. Intracellular ATP content, MTT reduction, and HSP70/HSP90 induction were studied; confocal, transmission electron microscopy, and light microscopy studies were also performed. For confocal analysis, a mitochondrial selective probe (MitoTracker Red CMXH2Ros) was used. Antioxidant activity of DMPS was also studied by the scavenging of the free radical 2, 2-diphenyl-1-picrylhydrazyl (DPPH) technique. A decrease of ATP content, an impaired ability to reduce tetrazolium, and dramatic changes on cellular and mitochondrial morphology, and energetic levels were found after either 6 or 9 h of HgCl(2) exposure. Increased expression of HSP90 and HSP70 were also seen. When OK cells were co-incubated with HgCl(2) and DMPS, cellular morphology, viability, intracellular ATP, and mitochondrial membrane potential were partially restored; a protective effect on mitochondrial morphology was also seen. DMPS also showed potent antioxidant activity in vitro. Mitochondrial protection could be the cellular mechanism mediated by DMPS in OK cells exposed to a toxic concentration of HgCl(2). PMID- 17131098 TI - Functionality of the beta/six site-specific recombination system in tobacco and Arabidopsis: a novel tool for genetic engineering of plant genomes. AB - The beta recombinase is a member of the prokaryotic site-specific serine recombinases (invertase/resolvase family), which in the presence of a DNA bending cofactor can catalyse DNA deletions between two directly oriented 90-bp six recombination sites. We have examined here whether the beta recombinase can be expressed in plants and whether it displays in planta its specific catalytic activity excising DNA sequences that are flanked by six sites. In plant protoplasts, the enzyme could be expressed as a GFP-beta recombinase fusion which can localise to the cell nucleus. Beta recombinase stably expressed in tobacco plants can catalyse deletion of a spacer region that is flanked by directly oriented six sites and has been placed between promoter and a GUS reporter gene (preventing GUS expression). In transient transformation experiments, beta recombinase-mediated elimination of the spacer results in transcriptional induction of the GUS gene. Similarly, beta recombinase in stably double transformed Arabidopsis plants deletes specifically the spacer region of a reporter construct that has been incorporated into the genome. In the segregating T1 generation, plants were identified that contain exclusively the recombined reporter construct. In summary, our results demonstrate that functional / recombinase can be expressed in plants and that the enzyme is suitable to precisely eliminate undesired sequences from plant genomes. Therefore, the beta/six recombination system (and presumably related recombinases) may become an attractive tool for plant genetic engineering. PMID- 17131099 TI - [Short bowel syndrome]. AB - Short bowel syndrome (SBS) is defined in adults as a malabsorption disorder as a result of shortening the bowel to <200 cm. The severity of symptoms is less dependent on the amount of residual intestine than on the anatomical position of the resected bowel, the type of operative reconstruction, and the type and quality of nutritional, medical, and surgical treatment. Numerous complications and deficiency symptoms are associated with SBS. The extent of deficient nutrition should be determined. The need to create accesses for enteral and parenteral delivery, to supply supplementation as needed, perform pharmacological therapy, and in individual cases surgical treatment all necessitate a broad knowledge of nutritional medicine. The goals of therapy are correction and prevention of malnourishment, restoration of a normal nutritional status, and the normal thriving of children. Complications should be avoided, particularly those problems associated with parenteral nutrition. The frequency of diarrhea should be reduced. Overall, the aim is to achieve an optimized quality of life. PMID- 17131100 TI - [Shoulder prostheses]. AB - The design of shoulder prostheses has been developed through four generations which mirror adaptation to our increasing knowledge of the biomechanics of the shoulder joint. Modern shoulder prostheses are adapted to the size, inclination, posterior offset, and retrotorsion of the shoulder. The main reasons for implantation of a shoulder prosthesis are primary osteoarthritis, posttraumatic and rheumatoid arthritis, avascular necrosis, instability arthritis and cuff defect arthropathy. Typical implants are cup prostheses for surface replacement, anatomical stem prostheses, and reverse prostheses. Total prostheses are functionally better as soon as the arthritis involves the glenoid, whereas hemiprostheses should be preferred as long as the glenoid is intact. The stem is mostly cemented, whereas in younger patients with good bone quality a cementless stem may be used. Cemented glenoids may be considered as standard. PMID- 17131101 TI - [Neurological Reha-Score. An instrument to measure outcome and expenditure of neurologic rehabilitation]. AB - BACKGROUND: A new measurement instrument was developed to combine common scores with the parameter of effort for rehabilitation, the latter being a control instrument for medical professionals and rehabilitational health organisations. METHODS: The Neurological Rehabilitation Score (NRS) contains 61 items (including the ten from the Barthel Index) resulting in scores of 0 (worst patient condition) to 600 (best). The items are grouped into seven categories: activities of daily life (13), mobility (8), communication and social skills (7), arm and hand function (7), orientation and cognition skills (10), strength and pain (8), and coping and miscellaneous (8). We investigated 8,139 patients and determined reliability (kappa statistic) and responsivity (standardised response means). In 100 consecutive patients, we also measured the Functional Independence Measure (FIM) and Scandinavian Stroke Scale (SSS) to address validity. RESULTS: With an average kappa value of 0.86 (range 0.62-0.98), the NRS showed excellent inter rater reliability. There was a good correlation between NRS, FIM, and SSS. Response was good at 0.84. The ceiling effect was also less than 1%, compared to 17% in the Barthel Index. CONCLUSION: The NRS was shown to be effective for evaluation of outcome and expenditure in neurological rehabilitation. PMID- 17131102 TI - [Guillain-Barre syndrome and inadequate antidiuretic hormone secretion syndrome]. AB - Several neurologic disorders including Guillain-Barre syndrome (GBS) are associated with hyponatremia. Hyponatremia and its overly fast correction have major implications to the course of the underlying neurologic disease. We report a case of GBS complicated by hyponatremia secondary to the development of inadequate antidiuretic hormone secretion syndrome. Differential diagnosis, pathophysiology, and therapeutic approach of hyponatremia in association with GBS are discussed. PMID- 17131103 TI - Detection and quantitation of colored deposit-forming Meiothermus spp. in paper industry processes and end products. AB - Colored biofilms cause problems in paper industry. In this work we used real-time PCR to detect and to quantitate members of the genus Meiothermus from the process samples and end products from 24 machines manufacturing pulp, paper and board in four countries. The results obtained from 200 samples showed the importance of members of the genus Meiothermus as ubiquitous biofoulers in paper machines. This genus was the dominant biofouler in some mills. From < or =10(4) to 10(11) copies of Meiothermus 16S rRNA genes were found per gram of process deposit (wet weight). Meiothermus spp. were found in paper and board products with colored defects and connection between deposit-forming microbes and end-product spots was shown. 16S rRNA gene sequences of 29 biofilm producing bacterial isolates from different mills were determined. Based on sequence data, 25 of the isolates were assigned to the genus Meiothermus, with Meiothermus silvanus and M. ruber as the most frequent species. PMID- 17131104 TI - Improvement of sordarin production through process optimization: combining traditional approaches with DOE. AB - BMS-353645, also known as sordarin, was of interest based on its activity against pathogenic fungi. The objective of these studies was to provide high quality starting substrate for chemical modification aimed at further improving biological activity, with particular interest in the inhibition of Aspergillus. In the work presented here, Design of Experiments, or DOE, was successfully combined with traditional approaches to significantly improve sordarin yields in fermentation flasks. Overall, yields were increased 25-fold from <100 microg/g to as high as 2,609 microg/g in flasks through the use of various medium and conduction changes supplemented with DOE. The improved process was then successfully scaled to pilot plant tanks with the best batch producing 2,389 microg/g sordarin at the 250-l scale. PMID- 17131105 TI - Marker-assisted selection and evaluation of the QTL for stigma exsertion under japonica rice genetic background. AB - Stigma exsertion is one of the important traits which contribute to the efficient improvement of commercial seed production in hybrid rice. In order to understand the genetic factors involved in the stigma exsertion of an indica variety--IR24- a QTL analysis was conducted using the F2 population between a japonica variety- Koshihikari--and a breeding line showing exserted stigma selected from the backcross population between IR24 as a donor and japonica varieties. As a result, a highly significant QTL (qES3), which had been predicted in the recombinant inbred population of IR24, was confirmed at the centromeric region on chromosome 3. qES3 increases about 20% of the frequency of the exserted stigmas at the IR24 allele and explains about 32% of the total phenotypic variance. A QTL near isogenic line for qES3 increased the frequency of the exserted stigma by 36% compared to that of Koshihikari in a field evaluation, which suggests that qES3 is a promising QTL for the development of a maternal line for hybrid rice. PMID- 17131106 TI - Identification of QTL and association of a phytoene synthase gene with endosperm colour in durum wheat. AB - The yellow colour of durum wheat (Triticum turgidum L. var durum) semolina is due in part to the presence of carotenoid pigments found in the endosperm and is an important end-use quality trait. We hypothesized that variation in the genes coding for phytoene synthase (Psy), a critical enzyme in carotenoid biosynthesis, may partially explain the phenotypic variation in endosperm colour observed among durum cultivars. Using rice sequence information, primers were designed to PCR clone and sequence the Psy genes from Kofa (high colour) and W9262-260D3 (medium colour) durum cultivars. Sequencing confirmed the presence of four Psy genes in each parent, corresponding to a two member gene family designated as Psy1-1, Psy1 2 and Psy2-1 and Psy2-2. A genetic map was constructed using 155 F1-derived doubled haploid lines from the cross W9262-260D3/Kofa with 194 simple sequence repeat and DArT markers. Using Psy1-1 and Psy2-1 allele-specific markers and chromosome mapping, the Psy1 and Psy2 genes were located to the group 7 and 5 chromosomes, respectively. Four quantitative trait loci (QTL) underlying phenotypic variation in endosperm colour were identified on chromosomes 2A, 4B, 6B, and 7B. The Psy1-1 locus co-segregated with the 7B QTL, demonstrating an association of this gene with phenotypic variation for endosperm colour. This work is the first report of mapping Psy genes and supports the role of Psy1-1 in elevated levels of endosperm colour in durum wheat. This gene is a target for the further development of a molecular marker to enhance selection for endosperm colour in durum wheat breeding programs. PMID- 17131107 TI - Amperometric hydrogen peroxide sensor based on a sol-gel-derived ceramic carbon composite electrode with toluidine blue covalently immobilized using 3 aminopropyltrimethoxysilane. AB - A carbon composite amperometric hydrogen peroxide sensor has been developed using a sol-gel technique. Toluidine blue (TB), which acts as the redox mediator, was covalently immobilized via glutaraldehyde crosslinking with an organically modified silane, namely 3-aminopropyltrimethoxysilane (APTMOS). Methyltrimethoxysilane (MTMOS) was used as the additional monomer; this controls the hydrophobicity of the electrode surface, thus limiting the wettability. The immobilization of TB within the sol-gel matrix was confirmed with FTIR studies. The sol-gel mixture containing TB immobilized in APTMOS and MTMOS was mixed with graphite powder in order to prepare the carbon composite electrode. The electrode was characterized using voltammetric techniques and its electrocatalytic activity for the reduction of hydrogen peroxide was also studied. The carbon composite electrode has the advantage of sensing H(2)O(2) at a lower potential and with a higher sensitivity, and interferences due to ascorbic acid, uric acid and acetaminophen were greatly minimized. The linear range for the determination of H(2)O(2) extends from 5.37 x 10(-6) to 6.15 x 10(-3) M, with a correlation coefficient of 0.9981. The detection limit was found to be 2.15 x 10(-6) M. The covalent immobilization of TB effectively prevents the leakage of the water soluble mediator during measurements. The modified electrode, aside from electrocatalyzing the reduction of H(2)O(2), exhibits distinct advantages in terms of surface renewal in the event of surface fouling, as well as simple preparation, good chemical and mechanical stability, and good reproducibility. PMID- 17131108 TI - Beyond blood sugar: the potential of NMR-based metabonomics for type 2 human diabetes, and the horse as a possible model. AB - Metabonomic analysis is a powerful tool for identifying and characterizing metabolic disorders, for example type 2 diabetes and the metabolic syndrome. Nuclear magnetic resonance (NMR) spectroscopy is an essential tool for such analysis, with special benefits. The review assesses the current status and potential of NMR-based metabonomics of type 2 diabetes. The horse is proposed as a possible model for studying this condition and disease. Some examples are shown of horse blood analyses by NMR. PMID- 17131109 TI - On-line monitoring of pH junctions in capillary electrophoresis using Fourier transform infrared spectrometry. AB - Fourier transform mid-infrared spectroscopic detection is proposed as an on-line detection technique for the study of on-line preconcentration processes in capillary electrophoresis (CE). The molecule-specific information contained in mid-IR spectra can be used to directly determine the chemical compositions of individual zones and their boundaries. This paper reports on pH junctions employed in myoglobin analysis. On-line mid-IR detection allowed the shape of the sample peak to be monitored as well as the chemical compositions of the surrounding zones. From this information it was possible to obtain detailed insights into the actual chemical compositions of the individual zones governing the efficiency of the preconcentration technique applied. The principle of measurement outlined here can therefore also be regarded as a promising one for investigating other on-line preconcentration techniques, like stacking, sweeping, and pH junction-sweeping among others. PMID- 17131110 TI - Pharmaceuticals and personal care products in biosolids/sewage sludge: the interface between analytical chemistry and regulation. AB - Modern sanitary practices result in large volumes of human waste, as well as domestic and industrial sewage, being collected and treated at common collection points, wastewater treatment plants (WWTPs). In recognition of the growing use of sewage sludge as fertilizers and soil amendments, and the scarcity of current data regarding the chemical constituents in sewage sludge, the US National Research Council (NRC) in 2002 produced a report on sewage sludge. Among the NRC's recommendations was the need for investigating the occurrence of pharmaceuticals and personal care products (PPCPs) in sewage sludge. PPCPs are a diverse array of non-regulated contaminants that had not been studied in previous sewage sludge surveys but which are likely to be present. The focus of this paper will be to review the current analytical methodologies available for investigating whether pharmaceuticals are present in WWTP-produced sewage sludge, to summarize current regulatory practices regarding sewage sludge, and to report on the presence of pharmaceuticals in sewage sludge. PMID- 17131111 TI - A microfluidic system for evaluation of antioxidant capacity based on a peroxyoxalate chemiluminescence assay. AB - A microfluidic system incorporating chemiluminescence detection is reported as a new tool for measuring antioxidant capacity. The detection is based on a peroxyoxalate chemiluminescence (PO-CL) assay with 9,10-bis (phenylethynyl)anthracene (BPEA) as the fluorescent probe and hydrogen peroxide as the oxidant. Antioxidant plugs injected into the hydrogen peroxide stream result in inhibition of the CL emission which can be quantified and correlated with antioxidant capacity. The PO-CL assay is performed in 800-microm-wide and 800-microm-deep microchannels on a poly(dimethylsiloxane) (PDMS) microchip. Controlled injection of the antioxidant plugs is performed through an injection valve. Of the plant-food based antioxidants tested, beta-carotene was found to be the most efficient hydrogen peroxide scavenger (SAHP of 3.27x10(-3) micromol-1 L), followed by alpha-tocopherol (SAHP of 2.36x10(-3) micromol-1 L) and quercetin (SAHP of 0.31x10(-3) micromol-1 L). Although the method is inherently simple and rapid, excellent analytical performance is afforded in terms of sensitivity, dynamic range, and precision, with RSD values typically below 1.5%. We expect our microfluidic devices to be used for in-the-field antioxidant capacity screening of plant-sourced food and pharmaceutical supplements. PMID- 17131112 TI - Use of quantitative real-time RT-PCR to analyse the expression of some quorum sensing regulated genes in Pseudomonas aeruginosa. AB - P. aeruginosa living in biofilm populations sends out diffusive signalling molecules, called autoinducers, for example acylated homoserine lactone (AHL) or the P. aeruginosa quinolone signal (PQS). So far, two quorum-sensing systems, LasR and VsmR, have been identified in P. aeruginosa, both of which are required for all virulence determinants. The expression of specific genes involved in quorum-sensing regulatory mechanisms has been analysed with molecular biology methods. Real-time quantitative PCR is a highly sensitive and powerful technique for quantification of nucleic acids. Expression of the genes vsmR, lasI, and PA4296 was studied by use of reverse transcriptase and subsequent quantitative real-time PCR of the cDNAs. In parallel, expression of ribosomal 16S rRNA, used as a housekeeping gene that was constitutively expressed in all analyses, was also monitored. Biofilm was compared with planktonic bacteria, and in contrast to vsmR and Pa4296, the lasI gene was found to be down-regulated in biofilm. Extended experiments were run with synthetic signal molecules inducing regulated processes in bacterial populations. It was shown that the genes under investigation were up-regulated in mature biofilm in the presence of the signal molecule N-(3-oxododecanoyl)-L-homoserine lactone. PMID- 17131113 TI - Optimal inference explains dimension-specific contractions of spatial perception. AB - It is known that people misperceive scenes they see during rapid eye movements called saccades. It has been suggested that some of these misperceptions could be an artifact of neurophysiological processes related to the internal remapping of spatial coordinates during saccades. Alternatively, we have recently suggested, based on a computational model, that transsaccadic misperceptions result from optimal inference. As one of the properties of the model, sudden object displacements that occur in sync with a saccade should be perceived as contracted in a non-linear fashion. To explore this model property, here we use computer simulations and psychophysical methods first to test how robust the model is to close-to-optimal approximations and second to test two model predictions: (a) contracted transsaccadic perception should be dimension-specific with more contraction for jumps parallel to the saccade than orthogonal to it, and (b) contraction should rise as a function of visuomotor noise. Our results are consistent with these predictions. They support the idea that human transsaccadic integration is governed by close-to-optimal inference. PMID- 17131114 TI - Comparison of manual tracing versus a semiautomatic radial measurement method in temporal lobe MRI volumetry for pharmacoresistant epilepsy. AB - INTRODUCTION: The aim of this study was to test a modified radial semiautomated volumetry technique (radial divider technique, RDT) versus the manual volumetry technique (MVT) for proportionality of temporal subvolumes in 30 patients with drug-resistant temporal lobe epilepsy. METHODS: Included in the study were 30 patients (15 female, 15 male; mean age 39.6 years) with pharmacoresistant epilepsy (mean duration 26.6 years). MRI studies were performed preoperatively on a 1.5-T scanner. All image processing steps and volume measurements were performed using ANALYZE software. The volumes of six subregions were measured bilaterally; these included the superior temporal gyrus (STG), middle + inferior temporal gyrus (MITG), fusiform gyrus (FG), parahippocampal gyrus (PHG), amygdala (AM), and hippocampus (HP). Linear regression was used to investigate the relationship between the comparable subvolumes obtained with MVT and RDT. RESULTS: Very high correlations (R (2) >0.95) between RDT and MVT were observed for the STG + MITG and the STG + MITG + FG, but low correlations for the PHG subvolumes and the combined PHG + HP + AM subvolumes. These observations were independent of the side of the pathology and of hemisphere. CONCLUSION: The two measurement techniques provided highly reliable proportional results. This series in a homogeneous group of TLE patients suggests that the much quicker RDT is suitable for determining the volume of temporolateral and laterobasal temporal lobe compartments, of both the affected and the non-affected side and the right and left hemisphere. PMID- 17131115 TI - Stenting for vertebrobasilar dissection: a possible treatment option for nonhemorrhagic vertebrobasilar dissection. AB - INTRODUCTION: It has been reported that stent placement may improve compromised blood flow resulting from vertebrobasilar dissection. In this study the technical feasibility, safety, as well as short-term outcome of stent placement for the treatment of nonhemorrhagic vertebrobasilar dissection was retrospectively investigated. METHODS: Ten patients (eight men, two women; age range 36 to 45 years) with nonhemorrhagic vertebrobasilar dissection were treated by stenting. Nine lesions were located at the vertebral artery (VA) (one bilateral case) and two at the basilar artery. Seven patients presented with ischemic symptoms and three with headache. Among the nine VA dissections, eight lesions involved the posterior inferior cerebellar artery (PICA). Angiographic findings included abrupt or irregular vessel narrowing with aneurysmal dilatation in nine lesions and irregular bulbous aneurysmal dilatation in two lesions. RESULTS: Placement of a stent-within-a-stent was performed in six lesions and single stent in five lesions. Initial treatments were technically successful in all patients. Follow up was performed using digital subtraction angiography (six patients) or CT angiography (two patients). Successful occlusion or decreased contrast filling of the aneurysm sac was noted in six patients (seven lesions), increased aneurysm sac filling in one patient, and parent artery occlusion in one patient. PICA flow was preserved in all those with follow-up (1 week to 17 months). CONCLUSION: Stent placement is technically feasible and safe for the treatment of vertebrobasilar artery dissection, especially for preserving PICA and/or major perforating arteries. However, a study with a larger population and longer follow up is necessary for validation of the efficacy of this treatment modality. PMID- 17131116 TI - Diffusion-weighted imaging and proton MR spectroscopy in the characterization of acute disseminated encephalomyelitis. AB - INTRODUCTION: Acute disseminated encephalomyelitis (ADEM) is usually a monophasic illness characterized by multiple lesions involving gray and white matter. Quantitative MR techniques were used to characterize and stage these lesions. METHODS: Eight patients (seven males and one female; mean age 19 years, range 5 to 36 years) were studied using conventional MRI (T2- and T1-weighted and FLAIR sequences), diffusion-weighted imaging (DWI) and proton magnetic resonance spectroscopy (MRS). Apparent diffusion coefficient (ADC) values and MRS ratios were calculated for the lesion and for normal-appearing white matter (NAWM). Three patients were imaged in the acute stage (within 7 days of the onset of neurological symptoms) and five in the subacute stage (after 7 days from the onset of symptoms). RESULTS: ADC values in NAWM were in the range 0.7-1.24 x 10( 3) mm/s2 (mean 0.937 +/- 0.17 mm/s2). ADC values of ADEM lesions in the acute stage were in the range 0.37-0.68 x 10(-3) mm/s2 (mean 0.56 +/- 0.16 mm/s2) and 1.01-1.31 x 10(-3) mm/s2 (mean 1.24 +/- 0.13 mm/s2) in the subacute stage. MRS ratios were obtained for all patients. NAA/Cho ratios were in the range 1.1-3.5 (mean 1.93 +/- 0.86) in the NAWM. NAA/Cho ratios within ADEM lesions in the acute stage were in the range 0.63-1.48 (mean 1.18 +/- 0.48) and 0.29-0.84 (mean 0.49 +/- 0.22) in the subacute stage. The ADC values, NAA/Cho and Cho/Cr ratios were significantly different between lesions in the acute and subacute stages (P < 0.001, P < 0.027, P < 0.047, respectively). ADC values were significantly different between lesions in the acute (P < 0.009) and subacute stages (P < 0.005) with NAWM. In addition, NAA/Cho and Cho/Cr ratios were significantly different between lesions in the subacute stage and NAWM (P < 0.006, P < 0.007, respectively). CONCLUSION: ADEM lesions were characterized in the acute stage by restricted diffusion and in the subacute stage by free diffusion and a decrease in NAA/Cho ratios. Restricted diffusion and progressive decrease in NAA/Cho ratios may help in staging the disease. PMID- 17131117 TI - Computational study of coagulation factor VIIa's affinity for phospholipid membranes. AB - The interaction between the gamma-carboxyglutamic acid-rich domain of coagulation factor VIIa (FVIIa), a vitamin-K-dependent enzyme, and phospholipid membranes plays a major role in initiation of blood coagulation. However, despite a high sequence and structural similarity to the Gla domain of other vitamin-K-dependent enzymes with a high membrane affinity, its affinity for negatively charged phospholipids is poor. A few amino acid differences are responsible for this observation. Based on the X-ray structure of lysophosphatidylserine (lysoPS) bound to the Gla domain of bovine prothrombin (Prth), models of the Gla domain of wildtype FVIIa and mutated FVIIa Gla domains in complex with lysoPS were built. Molecular dynamics (MD) and steered molecular dynamics (SMD) simulations on the complexes were applied to investigate the significant difference in the binding affinity. The MD simulation approach provides a structural and dynamic support to the role of P10Q and K32E mutations in the improvement of the membrane contact. Hence, rotation of the Gly11 main chain generated during the MD simulation results in a hydrogen bond with Q10 side chain as well as the appearance of a hydrogen bond between E32 and Q10 forcing the loop harbouring Arg9 and Arg15 to shrink and thereby enhances the accessibility of the phospholipids to the calcium ions. Furthermore, the application of the SMD simulation method to dissociate C6 lysoPS from a series of Gla domain models exhibits a ranking of the rupture force that can be useful in the interpretation of the PS interaction with Gla domains. Finally, adiabatic mapping of Gla6 residue in FVIIa with or without insertion of Tyr4 confirms the critical role of the insertion on the conformation of the side chain Gla6 in FVIIa and the corresponding Gla7 in Prth. PMID- 17131118 TI - Superior anti-tumor protection and therapeutic efficacy of vaccination with allogeneic and semiallogeneic dendritic cell/tumor cell fusion hybrids for murine colon adenocarcinoma. AB - Cancer immunotherapy by dendritic cell (DC)/tumor cell fusion hybrids (DC/TC hybrids) has been shown to elicit potent anti-tumor effects via the induction of immune responses against multiple tumor-associated antigens. In the present study, we compared the anti-tumor effects of vaccinating Balb/c mice (H-2(d)) with CT26CL25 colon carcinoma cells that had been fused with either syngeneic DCs from Balb/c mice, allogeneic DCs from C57BL/6 mice (H-2(b)) or semiallogeneic DCs from B6D2F1 mice (H-2(b/d)). Preimmunization with either semiallogeneic or allogeneic DC/TC hybrids induced complete protection from tumor challenge, whereas mice preimmunized with syngeneic DC/TC hybrids were only partially protected (75% tumor rejection). The average number of pulmonary metastases after intravenous tumor injection decreased significantly following immunization with semiallogeneic or allogeneic DC/TC hybrids (8.3 +/- 7.9 or 16.3 +/- 3.5, mean +/- SD) relative to syngeneic DC/TC hybrids (67.8 +/- 6.3). These data demonstrate that vaccination with semiallogeneic DC/TC hybrids resulted in the greatest anti tumor efficacy. Anti-tumor effects showed by in vivo studies were virtually accomplished by the frequency of induced CTLs specific to both gp70 and beta galactosidase assessed by using pentameric assay. Among the fusion vaccines tested, semiallogeneic DC/TC hybrids induced the highest ratio of Th1 cytokine IFN-gamma to Th2 cytokine IL-10. In addition, allogeneic or semiallogeneic DC/TC hybrids elicited a significantly stronger NK activity than syngeneic DC/TC hybrids. These findings suggest that in clinical settings, DCs derived from a healthy donor (which are generally characterized as more semiallogeneic than allogeneic) may be more capable than autologous DCs of inducing promising anti tumor effects in vaccinations with DC/TC hybrids. PMID- 17131119 TI - International meeting "Immunotherapy of cancer: challenges and needs". AB - The main aims of the international meeting "Immunotherapy of Cancer: Challenges and Needs" were to review the state of the art of cancer immunotherapy and to identify critical issues which deserve special attention for promoting progress of research in this field, with a particular focus on the perspectives of clinical research. Novel concepts and strategies for identifying, monitoring and predicting effective responses to cancer immunotherapy protocols were presented, focused on the use of adjuvants (CpG oligonucleotides) or cytokines (IFN-alpha) to enhance the efficacy of cancer vaccines. Moreover, the possible advantages of using different types of dendritic cells (for active immunization strategies) or T cells (for adoptive immunotherapy protocols) were debated. A consensus was achieved on the need for enhancing the efficacy of cancer vaccines or adoptive cell immunotherapy by combining these strategies with other anti-cancer treatments, including chemotherapy. Finally, initiatives for promoting clinical research by establishing a strategic cooperation in the field of cancer immunotherapy based on the active participation of all the relevant actors, including public institutions responsible of Public Health, National Cancer Institutes, industry, representatives of regulatory bodies, and patients' organizations were proposed. PMID- 17131120 TI - Mannan-binding lectin (MBL) in women with tumours of the reproductive system. AB - Mannan-binding lectin (MBL) is an important factor of innate immunity contributing to the clearance of microorganisms. Recently, an antitumourigenic role of MBL has been suggested. We investigated mbl2 genotypes, MBL concentrations, and MBL-MASP-2 complex activity in patients with ovarian cancer. The expression of both mbl2 and masp-2 genes were investigated in ovarian tissue sections. Additionally, samples from patients with other malignant and benign tumours of the reproductive tract were tested. A significantly higher incidence of MBL deficiency/insufficiency-associated genotypes was found among patients with malignant disease compared to age-matched controls. Unexpectedly, no differences in median MBL level or MBL-MASP-2 complex activity were found between the groups. This was partly a reflection of higher MBL concentrations and MBL MASP-2 activity in cancer patients compared with healthy women carrying corresponding genotypes. MBL-specific mRNA expression was detected in several normal and malignant ovarian tissues, as well as in ovarian epithelial cell lines. Intracellular staining with MBL-specific antibodies demonstrated the presence of MBL in ovarian cell lines, and in normal as well as malignant ovarian tissue sections. In contrast, MASP-2-specific mRNA expression was detected only in the ovary tissues of patients with malignant disease. No significant changes in MBL concentration during 3 months of chemotherapy were noticed. MBL was detected in ascites and in the fluid of benign ovarian cysts. Our findings may reflect anti-tumourigenic activity of MBL protein which might suggest potential therapeutic application. However, it cannot be excluded that mbl-2 mutant alleles may be in linkage disequilibrium with an unidentified tumour susceptibility gene(s). PMID- 17131121 TI - In the FVB/N HER-2/neu transgenic mouse both peripheral and central tolerance limit the immune response targeting HER-2/neu induced by Listeria monocytogenes based vaccines. AB - Listeria monocytogenes-based vaccines for HER-2/neu are capable of breaking tolerance in FVB/N rat HER-2/neu transgenic mice. The growth of implanted NT-2 tumors, derived from a spontaneously occurring tumor in the FVB/N HER-2/neu transgenic mouse, was significantly slower in these mice following vaccination with a series of L. monocytogenes-based vaccines for HER-2/neu. Mechanisms of T cell tolerance that exist in these transgenic mice include the absence of functional high avidity anti-HER-2/neu CD8(+) T cells and the presence of CD4(+)CD25(+) regulatory T cells. The in vivo depletion of these regulatory T cells resulted in the slowing in growth of tumors even without the treatment of mice with an anti-HER-2/neu vaccine. The average avidities of responsive CD8(+) T cells to six of the nine epitopes in HER-2/neu we examined, four of which were identified in this study, are shown here to be of a lower average avidity in the transgenic mice versus wild type FVB/N mice. In contrast, the average avidity of CD8(+) T cells to three epitopes that showed the lowest avidity in the wild-type mice did not differ between wild type and transgenic mice. This study demonstrates the ability of L. monocytogenes-based vaccines to impact upon tolerance to HER-2/neu in FVB/N HER-2/neu transgenic mice and further defines some of the aspects of tolerance in these mice. PMID- 17131122 TI - Individual Vgamma2-Jgamma1.2+ T cells respond to both isopentenyl pyrophosphate and Daudi cell stimulation: generating tumor effectors with low molecular weight phosphoantigens. AB - Human Vgamma2Vdelta2 T cells exhibit T cell receptor-dependent, MHC-unrestricted recognition of antigen and play important roles in tumor and pathogen immunity. To characterize antigen recognition by the Vgamma2Vdelta2 TCR, we used the combined approach of spectratyping and CDR3 sequence analysis that measures changes in the TCR repertoire before and after stimulation with a phosphoantigen (isopentenyl pyrophosphate) or an irradiated tumor cell line (Daudi B lymphoma). Here we describe common Vgamma2 chains that are substantially involved in the response to both phosphoantigens and tumor cells. The recognition properties of common Vgamma2 chains explains the observation that Vgamma2Vdelta2 T cells expanded by phosphoantigen stimulation specifically recognize and kill some but not all tumor cell lines. Our studies further justify efforts to stimulate tumor immunity by administering low molecular weight phosphoantigens and boosting the frequency and tumor effector functions of circulating Vgamma2Vdelta2 T cells. PMID- 17131123 TI - Late onset of bortezomib-associated cutaneous reaction following herpes zoster. PMID- 17131124 TI - An update on the assessment of osteoporosis using radiologic techniques. AB - In this article, the currently available radiologic techniques for assessing osteoporosis are reviewed. Density measurements of the skeleton using dual X-ray absorptiometry (DXA) are clinically indicated for the assessment of osteoporosis and for the evaluation of therapies. DXA is the most widely used technique for identifying patients with osteoporosis. Quantitative computed tomography (QCT) is the only method, which provides a volumetric density. Unlike DXA, QCT allows for selective trabecular measurement and is less sensitive to degenerative diseases of the spine. The analysis of bone structure in conjunction with bone density is an exciting new field in the assessment of osteoporosis. High-resolution multi slice CT and micro-CT are useful tools for the assessment of bone microarchitecture. A growing literature indicates that quantitative ultrasound (QUS) techniques are capable of assessing fracture risk. Although the ease of use and the absence of ionizing radiation make QUS attractive, the specific role of QUS techniques in clinical practice needs further determination. Considerable progress has been made in the development of MR techniques for assessing osteoporosis during the last few years. In addition to relaxometry techniques, high-resolution MR imaging, diffusion MR imaging and in-vivo MR spectroscopy may be used to quantify trabecular bone architecture and mineral composition. PMID- 17131125 TI - Non-arteritic anterior ischaemic optic neuropathy: evaluation of the brain and optic pathway by conventional MRI and magnetisation transfer imaging. AB - The purpose of the study was to examine the brain and the visual pathway of patients with non-arteritic anterior ischaemic optic neuropathy (NAION) by using conventional MRI (cMRI) and volumetric magnetisation transfer imaging (MTI). Thirty NAION patients, aged 67.5 +/- 8.14 years, and 28 age- and gender-matched controls were studied. MTI was used to measure the magnetisation transfer ratio (MTR) of the chiasm and for MTR histograms of the brain. The presence of areas of white matter hyperintensity (WMH) was evaluated on fluid-attenuated inversion recovery (FLAIR) images. Area of the optic nerves (ONs) and volume of the chiasm were assessed, as were coronal short-tau inversion recovery (STIR) and MTI images, respectively. More areas of WMH were observed in patients (total 419; mean 14.4; SD 19) than in controls (total 127; mean 4.7; SD 5.7), P < 0.001. Area (in square millimetres) of the affected ONs, volume(in cubic millimetres) and MTR (in percent) of the chiasm (10.7 +/- 4.6), (75.8 +/- 20.2), (56.4 +/- 6.5), respectively, were lower in patients than in controls (13.6 +/- 4.3), (158.2 +/- 75.3) (62.1 +/- 6.2), respectively, P < 0.05. Mean MTR of brain histograms was lower in patients (53.0 +/- 8.0) than in controls (58.0 +/- 5.6), P < 0.05. NAION is characterised by decreased ON and chiasmatic size. The low MTR of the chiasm and brain associated with increased areas of WMH may be suggestive of demyelination and axonal damage due to generalised cerebral vascular disease. PMID- 17131126 TI - Prostate dynamic contrast-enhanced MRI with simple visual diagnostic criteria: is it reasonable? AB - The purpose of this study was to evaluate the accuracy of prostate cancer localization with simple visual diagnostic criteria using dynamic contrast enhanced (DCE) magnetic resonance imaging (MRI). A total of 46 consecutive patients with biopsy-proven prostate cancer underwent prostate 1.5 T MRI with pelvic phased-array coils before prostatectomy. Besides the usual T2-weighted sequences, a 30-s DCE sequence was acquired three times after gadoterate injection. On DCE images, all early enhancing lesions of the peripheral zone were considered malignant. In the central gland, only early enhancing lesions appearing homogeneous or invading the peripheral zone were considered malignant. Three readers specified the presence of cancer in 20 prostate sectors and the location of distinct tumors. Results were compared with histology; p < 0.05 was considered significant. For localization of cancer in the sectors, DCE imaging had a significantly higher sensitivity [logistic regression, odds ratio (OR): 3.9, p < 0.0001] and a slightly but significantly lower specificity (OR: 0.57, p < 0.0001). Of the tumors >0.3 cc, 50-60% and 78-81% were correctly depicted with T2-weighted and DCE imaging, respectively. For both techniques, the depiction rate of tumors >0.3 cc was significantly influenced by the Gleason score (most Gleason /=10 points or when a transfusion of at least three units of concentrated red blood cells is needed). The main outcome measures are: (1) indications for urgent surgery and results, (2) morbidity and mortality, and (3) relapse. RESULTS: There were 158 (92%) stable patients, and in 61% of these, the bleeding was identified via colonoscopy. Bleeding was identified using urgent colonoscopy in a higher percentage of patients compared to delayed colonoscopy (68% versus 14%; p < 0.001). Urgent surgery was indicated in 24 (14%) patients, and the approach was peri-anal in 5 (21%) patients and abdominal in the rest. Local intestinal resection was performed on the 15 patients in which bleeding was identified, whereas a subtotal colectomy was performed on the remaining 4 patients. The presence of hypotension (p = 0.001; 35 versus 10%) and etiology of LGIB (p < 0.001) are prognostic factors of urgent surgery. Morbidity was 6.4%, and mortality was 4.7%. The only morbidity or mortality risk factors detected were the presence of associated comorbidities (p = 0.008) and the need for urgent surgery (p = 0.002). The most frequent etiology was diverticulosis (25%). After a mean follow-up of 132 +/- 75 months, bleeding relapsed in 30% of patients. CONCLUSIONS: It is difficult to predict which patients are going to need urgent surgery in severe acute LGIB; only the presence of hypotension on arrival at the emergency ward would lead us to suspect a negative outcome for the hemorrhage. In severe acute LGIB, morbidity and mortality is high, and this is mainly due to the high level of associated comorbidity and the need for urgent surgery. It is necessary for strict hemodynamic monitoring of the patients at risk if we want to improve outcomes. The bleeding relapse rate is high in LGIB, although generally, it is not severe. PMID- 17131154 TI - New trends in the treatment of undifferentiated carcinomas of the thyroid. AB - INTRODUCTION: Malignant tumours of the thyroid are generally classified as either well-differentiated thyroid carcinoma, which is composed of papillary and follicular carcinoma, or undifferentiated/anaplastic thyroid carcinoma (ATC). ATC is not only the most lethal disease in the field of endocrine surgery but also one of the most aggressive tumours characterized by an almost invariable fatal outcome, which only very rarely exceeds a 1-year course. DISCUSSION: The impact of surgical resection in association with external beam radiation on ATC outcome has been extensively investigated also in studies based on multicentric database, and there is a general agreement on the significance of a complete resection of the tumour. It has been difficult up to now to collect data regarding chemotherapy adjuvant treatment. In spite of the lack of an extensive review about the results of this kind of treatment by itself or as part of a multimodal approach, it seems that among the several chemotherapy agents experienced, none proved to influence significantly ATC prognosis. Neither doxorubicin (the most commonly used) nor other drugs, such as cisplatin, bleomycin, fluorouracil or cyclophosphamide, showed any real efficacy in controlling the disease. CONCLUSION: The most recent development in this field seems to be represented by the possibility offered by PPARg agonists; even more promising might be the use of adenovirus-mediated p53 tumour suppressor gene therapy or BMP-7. All these new therapies need further confirmation coming from ongoing clinical trials such as those involving the use of vascular and growth factor-targeted agents. PMID- 17131155 TI - A clinical chameleon: postoperative hypoparathyroidism. AB - BACKGROUND: About 1,200 patients per year develop postoperative hypoparathyroidism alone in Germany. Many of those patients may be misdiagnosed as the symptoms of this disease may vary and can be atypical. PATIENT/RESULTS: As an example, we describe the first known case of an elderly patient with a long history of seizures as a complication of an undiagnosed chronic hypoparathyroidism after surgery of a pT4-esophageal carcinoma. The 63-year-old male patient underwent laryngo-hypopharyngo-esophagectomy with gastric transposition and partial thyroid resection for a proximal esophageal carcinoma in 1994. About half a year later, the patient developed for the first time a convulsive syncope. Misleading diagnoses were for years suspected metastasis formation and a dumping syndrome. The general physician of the patient called him the epilepsy man, while no cause of the seizures were found. More than a decade of years later, when the correct diagnosis was made, finally by determination of parathyroid hormone levels, the seizures of the patient were completely eliminated by calcium supplementation therapy. The patient's quality of life improved clearly in the following time. CONCLUSION: It is essential to consider chronic hypoparathyroidism in the differential diagnosis of patients with hypocalcemia who have undergone extended neck and proximal esophageal surgery before. In addition to that, it is mandatory to autotransplant parathyroids during the initial procedure which might be accidentally removed during surgery and to monitor parathyroid function in each patient in the further course postoperatively. PMID- 17131156 TI - Perioperative outcome in sarcoma surgery. AB - INTRODUCTION: Soft tissue sarcomas (STS) are rare tumors accounting for less than 1% of all malignancies. Although disease-specific surgical management is increasingly important, only few data are available for STS. Here, we analyze a single institution setting focusing on perioperative surgical and clinical parameters. METHODS: Prospectively gathered data of all adult patients undergoing surgery for STS including gastrointestinal stroma tumors (GIST) between October 2001 and October 2004. Patients undergoing only biopsy or ambulatory surgery were excluded. Statistical analysis was performed using SAS(R) software and patient's data from a computerized sarcoma registry. RESULTS: 159 patients with a median age of 60.2 years underwent a total of 179 operations. Three major sites of occurrence were notified: the visceral cavity (VIS) (36.3%), the retroperitoneum (RET) (31.3%), and the extremities (EXT) (27.4%). GIST (53.9%) were the most common type in the VIS, liposarcoma (62.5%) in the RET, and either liposarcoma (30.6%) or malignant fibrous histiocytoma (28.6%) in the EXT. Recurrence was treated in more than half of the patients with RET STS, and in almost one third of the EXT lesions, while primary occurrence dominated in the VIS. Median operation times in the VIS, RET, and EXT were 210, 240, and 120 min, respectively. Blood loss was 300, 500, and 50 ml for VIS, RET, and EXT operations. Morbidity was 26.2, 30.4, and 34.7% in VIS, RET, and EXT operations, respectively (reoperation rates were 9.4, 5.4, and 14.3%). Mortality was 1.5, 8.9, and 2.0% for VIS, RET, and EXT. Length of hospital stay in the groups was comparable. CONCLUSION: STS surgery of a single surgical unit contains predominantly VIS, RET, and EXT tumors. The STS subtype varies with location, as does length of operation, blood loss, morbidity, mortality, and reoperation rate. These data are helpful for planning the perioperative management of adult patients with STS and can be used for prognostic analyses. PMID- 17131157 TI - Rod in loop ileostomy: just an insignificant detail for ileostomy-related complications? AB - BACKGROUND AND AIMS: The aim of this prospective study was to validate a variant in the loop ileostomy construction to reduce peristomal pressure ulcers and, subsequently, the need of stoma therapist assistance and the frequency of changing the stoma appliance. PATIENTS AND METHODS: We have enrolled 33 consecutive patients who underwent two stage restorative proctocolectomies. The first consecutive 13 patients operated on had their ileostomies constructed with a standard rod. In the following 20 patients, we placed a 5.3-mm suction catheter tube closed with a stitch to form a "ring" and without any stitches fixing it to the skin. RESULTS: In the "ring" rod group 40% of patients did not report any complication compared to the 8% of patients in the standard rod group (p = 0.046). Pressure ulcers were absent in this group, while it affected 61% of the patients in the standard rod group (p < 0.001). Patients in the "ring" rod group needed significantly less assistance time by the stoma therapist (p < 0.01) and required significantly fewer stoma appliance changes (p < 0.01). In our institution, the overall cost for the complete management of a standard rod ileostomy was 73.16 (29.83-130.49) euro compared to 46.65 (23.15-93.48) euro for a "ring" rod ileostomy (p = 0.002). CONCLUSIONS: The adoption of a "ring" rod configuration led to an elimination of pressure ulcers due to the rigid rod, a shorter time requirement for stoma care and a decreased number of appliances required and was subsequently associated with lower costs of assistance. A tighter fitting around the ileostomy that avoided stool infiltration improved the practical management of the stoma with a "ring" rod. PMID- 17131158 TI - Identification of differentially expressed genes in the developing antler of red deer Cervus elaphus. AB - Understanding the molecular mechanisms underlying bone development is a fundamental and fascinating problem in developmental biology, with significant medical implications. Here, we have identified the expression patterns for 36 genes that were characteristic or dominant in the consecutive cell differentiation zones (mesenchyme, precartilage, cartilage) of the tip section of the developing velvet antler of red deer Cervus elaphus. Two major functional groups of these genes clearly outlined: six genes linked to high metabolic demand and other five to tumor biology. Our study demonstrates the advantages of the antler as a source of mesenchymal markers, for distinguishing precartilage and cartilage by different gene expression patterns and for identifying genes involved in the robust bone development, a striking feature of the growing antler. Putative roles for "antler" genes that encode alpha-tropomyosine (tpm1), transgelin (tagln), annexin 2 (anxa2), phosphatidylethanolamine-binding protein (pebp) and apolipoprotein D (apoD) in intense but still controlled tissue proliferation are discussed. PMID- 17131159 TI - Both sense and antisense strands of the LTR of the Schistosoma mansoni Pao-like retrotransposon Sinbad drive luciferase expression. AB - Long terminal repeat (LTR) retrotransposons, mobile genetic elements comprising substantial proportions of many eukaryotic genomes, are so named for the presence of LTRs, direct repeats about 250-600 bp in length flanking the open reading frames that encode the retrotransposon enzymes and structural proteins. LTRs include promotor functions as well as other roles in retrotransposition. LTR retrotransposons, including the Gypsy-like Boudicca and the Pao/BEL-like Sinbad elements, comprise a substantial proportion of the genome of the human blood fluke, Schistosoma mansoni. In order to deduce the capability of specific copies of Boudicca and Sinbad LTRs to function as promotors, these LTRs were investigated analytically and experimentally. Sequence analysis revealed the presence of TATA boxes, canonical polyadenylation signals, and direct inverted repeats within the LTRs of both the Boudicca and Sinbad retrotransposons. Inserted in the reporter plasmid pGL3, the LTR of Sinbad drove firefly luciferase activity in HeLa cells in its forward and inverted orientation. In contrast, the LTR of Boudicca did not drive luciferase activity in HeLa cells. The ability of the Sinbad LTR to transcribe in both its forward and inverted orientation represents one of few documented examples of bidirectional promotor function. PMID- 17131160 TI - Serum trace element concentrations in children with chronic renal failure. PMID- 17131161 TI - Growth impairment shows an age-dependent pattern in boys with chronic kidney disease. AB - The impact of chronological age on longitudinal body growth from early childhood through adolescence using detailed anthropometric methods has not yet been studied in children with chronic kidney disease (CKD). We have evaluated growth failure by measuring four components of linear growth: body height (HT), sitting height (SHT), arm length (AL) and leg length (LL). Data were prospectively collected for up to 7 years on 190 boys (3-21 years old) with congenital or hereditary CKD (all had developed at least stage 2 CKD by the age of 10 years). Patients showed the most severe growth failure in early childhood, followed by an acceleration in growth in pre-puberty, a slowing-down of growth at puberty, as expected, and thereafter a late speeding-up of growth until early adulthood. This pattern was observed irrespective of the degree of CKD and different treatment modalities, such as conservative treatment, recombinant human growth hormone (rhGH) therapy or transplantation. LL showed the most dynamic growth changes of all the parameters evaluated and emerged as the best indicator of statural growth in children with CKD. A specific age-dependent pattern of physical growth was identified in pediatric male CKD patients. This growth pattern should be considered in the evaluation of individual growth and the assessment of treatment efficacy such as rhGH therapy. PMID- 17131162 TI - Clinical outcomes of childhood lupus nephritis: a single center's experience. AB - This study retrospectively reviewed the medical records of children with lupus nephritis (LN) who were treated at Seoul National University Children's Hospital from 1986 to 2005 (mean duration 8.3+/-4.4 years). The records of 77 children (22 male and 55 female) were examined. The mean age at diagnosis was 11.9+/-3.0 years. The initial biopsy results revealed a WHO class IV classification for 60 (88.2%) of 68 biopsy proven cases. Of 77 patients, 67 (87.0%) responded initially to the high-dose corticosteroids with or without additional immunosuppressive therapy. Of the initial responders (67), 30 (44.8%) experienced at least one episode of proteinuric (24) or nephritic (6) flare. Thirteen patients (16.9%) progressed to either chronic renal failure (CRF) or end-stage renal disease (ESRD). Six (7.8%) patients died. A Kaplan-Meier estimate of patient survival and CRF-free survival rate was 95.4% and 88.7% at 5 years and 91.8% and 74.7% at 10 years, respectively. Multivariate analysis for class IV LN revealed male gender (P=0.029), initial hypertension (P=0.001) and absence of remission (P=0.002) to be prognostic factors predicting CRF. Glomerulosclerosis of 10% or more (P=0.005), nephritic flare (P=0.011), and presence of anti-phospholipid antibody (P=0.017) or syndrome (P=0.004) were also found to be independent risk factors for CRF. Cyclophosphamide pulse therapy failed to demonstrate superiority over other combined immunosuppressants used for the treatment of diffuse proliferative LN. PMID- 17131163 TI - Hybridization analysis of D4Z4 repeat arrays linked to FSHD. AB - Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant disease involving shortening of D4Z4, an array of tandem 3.3-kb repeat units on chromosome 4. These arrays are in subtelomeric regions of 4q and 10q and have 1 100 units. FSHD is associated with an array of 1-10 units at 4q35. Unambiguous clinical diagnosis of FSHD depends on determining the array length at 4q35, usually with the array-adjacent p13E-11 probe after pulsed-field or linear gel electrophoresis. Complicating factors for molecular diagnosis of FSHD are the phenotypically neutral 10q D4Z4 arrays, cross-hybridizing sequences elsewhere in the genome, deletions including the genomic p13E-11 sequence and part of D4Z4, translocations between 4q and 10q D4Z4 arrays, and the extremely high G + C content of D4Z4 arrays (73%). In this study, we optimized conditions for molecular diagnosis of FSHD with a 1-kb D4Z4 subfragment probe after hybridization with p13E-11. We demonstrate that these hybridization conditions allow the identification of FSHD alleles with deletions of the genomic p13E-11 sequence and aid in determination of the nonpathogenic D4Z4 arrays at 10q. Furthermore, we show that the D4Z4-like sequences present elsewhere in the genome are not tandemly arranged, like those at 4q35 and 10q26. PMID- 17131164 TI - A descriptive study of injuries in a pediatric population of North-Eastern Italy. AB - Childhood injuries are a major public health problem in Italy. From a study conducted in 1984, injury rates were found to be higher in Trieste, in the north east of the country, than in other Italian areas. We conducted a new study to evaluate whether injury rates and patterns have changed in Trieste. There are three emergency rooms (ER) in Trieste. We collected and analyzed information on all injured children 0-16 years of age attending these ER in 2003 (child population 0-16 years of age was 28,000). We calculated the annual injury risk overall and by age. We described characteristics of the children (age, sex) and injuries (place, cause, type, affected body part, severity). 5,928 injured children attended the ER, and the annual injury risk was 21.5%. The home was the most frequent place where injuries occurred, especially among the youngest children. The most commonly injured body parts were the limbs and, among the youngest children, the head and face. Approximately 20% of children had moderate to severe injuries (AIS>1), and less then 3% were admitted to the hospital. In comparison to the previous study, there have been no significant changes in the annual risk of childhood injury and in the injury patterns. On the contrary, we observed a dramatic reduction in the frequency of hospitalization, which is probably attributable to the recent implementation of short observation and to the improvement of diagnostic/therapeutic paths in the ER. In conclusion, childhood injuries are still a relevant public health problem in this Italian area and new efforts are needed to prevent them. PMID- 17131165 TI - Historical perspective of the word "sepsis". PMID- 17131167 TI - Academic medical centers, private industry, and clinical trials: how do we achieve fairness, objectivity, and balance? PMID- 17131168 TI - A comparative study of water perfusion catheters and microtip transducer catheters for urethral pressure measurements. AB - The aim of this study was to compare the maximum urethral closure pressure (MUCP) measures with two different techniques: water perfused catheter and microtip transducer catheters with respect to reproducibility and comparability for urethral pressure measurements. Eighteen women with stress urinary incontinence had repeat static urethral pressure profilometry on a different day using a dual microtip transducer and water perfused catheter (Brown and Wickham). The investigators were blinded to the results of the other. The microtip measurements were taken in the 45 degrees upright sitting position with the patient at rest at a bladder capacity of 250 ml using an 8 Fr Gaeltec double microtip transducer withdrawn at 1 mm/s, and the transducer was orientated in the three o'clock position. Three different measures were taken for each patient. Three water perfusion measurements were performed with the patient at rest in the 45 degrees upright position at a bladder capacity of 250 ml using an 8 Fr BARD dual lumen catheter withdrawn at 1 mm/s. The mean water perfusion MUCP measure was 26.1 cm H2O, significantly lower than the mean microtip measure of 35.7 cm H2O. The correlation coefficient comparing each water perfusion measurement with the other water perfusion measures in the same patient was excellent, at 0.95 (p = 0.01). Correlation coefficient comparing each microtip measure with the other microtip measure in the same patient was also good, ranging from 0.70 to 0.80. This study confirms that both water perfusion catheters and microtip transducers have excellent or very good reproducibility with an acceptable intraindividual variation for both methods. PMID- 17131169 TI - Nocturnal polyuria and nocturia relief in patients treated with solifenacin for overactive bladder symptoms. AB - The objectives of this study are to determine whether patients with overactive bladder (OAB) and nocturia achieved relief of their nighttime voiding symptoms when treated with solifenacin and whether having nocturnal polyuria (NP) affected that response. The patients pooled from four phase III clinical trials were evaluated for reductions in nocturia episodes after treatment with solifenacin (5 or 10 mg) or placebo. A second analysis was performed in patients with and without NP. The patients treated with solifenacin experienced statistically significant reductions in nocturia episodes; median reductions were -35.5% for 5 mg of solifenacin and -36.4% for 10 mg of solifenacin compared with -25.0% for placebo, and significantly more patients treated with solifenacin vs placebo achieved a mean nocturic frequency of Gln], Hoshida [Glu-43 (beta)-->Gln] and alpha(2)beta (2) (T87Q) mutations as the prototype of three distinct classes of contact sites of deoxy HbS fiber. Binary mixture experiments established that beta(A)-chain with the Thr-87 (beta)-->Gln mutation is as potent as the gamma chain of HbF (alpha(2)gamma(2)) in inhibiting polymerization. On combining the influence of Le Lamentin mutation with that of beta (2) (T87Q) mutations; the net influence is only partial additivity. On the other hand, in binary mixture studies, combined influence of Hoshida mutation with that of beta (2) (T87Q) mutations is synergistic. Besides, a significant level of synergistic complementation is also seen when the Le Lamentin and Hoshida mutations are combined in HbS (symmetrical tetramers). Le Lamentin and Hoshida mutation introduced into the cis-dimer of the asymmetric hybrid tetramer completely neutralizes the Val-6 (beta) dependent polymerization. Accordingly, we propose that combining the perturbation of intra-double strand contact site with that of an inter-double strand contact site exhibit synergy when they are present in two different chains of the alphabeta dimer. A comparison of the present results with that of the earlier studies suggest that when the two contact site perturbations are from the same sub-unit of the alphabeta dimer only partial additivity is observed. The map of interaction linkage of the contact site mutations exposes new strategies in the design of novel anti-sickling Hbs for the gene therapy of sickle cell disease. PMID- 17131195 TI - Differential scanning calorimetry of a metalloprotein under controlled metal-ion activity. AB - In order to investigate the thermodynamics of the unfolding of metalloproteins, the thermal denaturation of bovine alpha-lactalbumin (BLA), a typical calcium binding protein, was investigated under a wide variety of calcium ion activities by means of differential scanning calorimetry. The excess heat capacity obtained as above is composed of those of the following three reactions: (i) the release of a calcium ion from holo-BLA; (ii) the capture of the released calcium ion by the chelating reagent; and (iii) the denaturation of native apo-BLA. The results indicated that the presence of the chelating reagent had a remarkable effect on the apparent enthalpy change for the denaturation of holo-BLA. On the other hand, the influence of the chelator on the heat capacity change was shown to be negligible. Because the denaturation reaction of holo-BLA includes Reactions (i) and (iii), it had to be handled as a three-state reaction. Such an investigation of the unfolding has been scarcely found that the activity of the metal ion is controlled precisely in wide range. PMID- 17131196 TI - Geographic access to family planning facilities and the risk of unintended and teenage pregnancy. AB - OBJECTIVES: This study tested the hypotheses that greater geographic access to family planning facilities is associated with lower rates of unintended and teenage pregnancies. METHODS: State Pregnancy Risk Assessment Monitoring System (PRAMS) and natality files in four states were used to locate unintended and teenage births, respectively. Geographic availability was measured by cohort travel time to the nearest family planning facility, the presence of a family planning facility in a ZIP area, and the supply of primary care physicians and obstetric-gynecologists. RESULTS: 83% of the PRAMS cohort and 80% of teenagers lived within 15 min or less of a facility and virtually none lived more than 30 min. Adjusted odds ratios did not demonstrate a statistically significant trend to a higher risk of unintended pregnancies with longer travel time. Similarly there was no association with unintended pregnancy and the presence of a family planning facility within the ZIP area of maternal residence, or with the supply of physicians capable of providing family planning services. Both crude and adjusted relative rates of teenage pregnancies were significantly lower with further distance from family planning sites and with the absence of a facility in the ZIP area of residence. In adjusted models, the supply of obstetricians gynecologists and primary care physicians was not significantly associated with decreased teen pregnancies. CONCLUSIONS: This study found no relationship between greater geographic availability of family planning facilities and a risk of unintended pregnancies. Greater geographic availability of family planning services was associated with a higher risk of teenage pregnancy, although these results may be confounded by facilities locating in areas with greater family planning needs. PMID- 17131197 TI - Accessibility of family planning services: impact of structural and organizational factors. AB - OBJECTIVES: This study sought to determine whether selected structural and organizational characteristics of publicly available family planning facilities are associated with greater availability. METHODS: A survey was sent to 726 publicly available family planning facilities in four states. These included local health departments, federally qualified health centers (FQHC), Planned Parenthood sites, hospital outpatient departments, and freestanding women's health centers. Usable responses were obtained from 526 sites for a response rate of 72.5%. Availability variables included the provision of primary care services; the contraceptives offered; professional staffing; scheduling, waiting time, and transportation; and cultural congruence and competency. The structural and organizational variables were state, type of organization, and funding source. RESULTS: Some states were more likely to offer emergency contraception while others were more likely to have weekend hours. FQHCs were most likely to provide primary care and Planned Parenthood sites most likely to offer emergency contraception. Title X funding was associated with increased likelihood of providing emergency contraception and staffing by midlevel practitioners and registered nurses. CONCLUSIONS: This study found that availability varied by structural and organizational variables, many of which are determined by federal and state policies. Revising some of these policies might increase utilization of family planning facilities. PMID- 17131198 TI - The sex ratio of older siblings in non-right-handed homosexual men. AB - This study tested the prediction, based on prior research, that non-right-handed homosexual men will report fewer than expected older brothers. Participants were 2486 heterosexual and homosexual, right-handed and non-right-handed, male and female adults, representing five samples collected for various projects by the second author. Data on sibship composition, sexual orientation, and hand preference were gathered in the original research using on-line (Internet) or self-administered paper-and-pencil questionnaires. The non-right-handed homosexual men reported 83 older brothers per 100 older sisters, which was significantly lower than the human sex ratio of 106 live-born males per 100 live born females. In contrast, the right-handed homosexual men reported 125 older brothers per 100 older sisters, which was significantly higher than the expected ratio. One possible explanation of these results is that older brothers increase the odds of homosexuality in right-handed males but decrease the odds of homosexuality in non-right-handed males. A second possibility is that older brothers decrease the probability that non-right-handed homosexual males will be represented in survey research. The latter scenario could arise if the combination of some biological factor associated with older brothers and some biological factor associated with non-right-handedness is so toxic that it kills the fetus or predisposes the individual to a condition (e.g., mental retardation, major mental illness) that makes him less likely to be available for research recruitment at Gay Pride parades (etc.) than other members of the gay community. PMID- 17131199 TI - The association between conduct problems and the initiation and progression of marijuana use during adolescence: a genetic analysis across time. AB - The present study used a prospective, longitudinal design to investigate genetic and environmental influences on the association between earlier conduct problems and the initiation and progression of marijuana use during adolescence. Parent- and teacher-reported conduct problems assessed at Time 1 (1996) and self-reported marijuana use assessed at Time 2 (2004) were available for 1088 adolescent twin pairs participating in the Cardiff Study of All Wales and North West of England Twins (CaStANET). Using a novel approach to the modeling of initiation and progression dimensions in substance use, findings suggested that the initiation of marijuana use in adolescence was influenced by genetic, common and unique environmental factors. The progression (or frequency) of marijuana use was influenced by genetic and unique environmental factors. Findings for conduct problems indicated that while the presence or absence of conduct problems was largely heritable, the relative severity of conduct problems appeared to be more strongly environmentally influenced. Multivariate model fitting indicated that conduct problems in childhood and early adolescence made a small but significant contribution to the risk for marijuana use 8 years later. PMID- 17131200 TI - Genetic and structural analysis of the basolateral amygdala complex in BXD recombinant inbred mice. AB - The amygdala integrates and coordinates emotional and autonomic responses. The genetics that underlie variation in amygdala structure may be coupled to variation in levels of aggression, fear, anxiety, and affiliated behaviors. We systematically quantified the volume and cell populations of the basolateral amygdala complex (BLAc) across 35 BXD recombinant inbred (RI) lines, the parental strains--C57BL/6J (B6) and DBA/2J (D2)--and F1 hybrids (n cases=199, bilateral analysis). Neuron number and volume vary 1.7- to 2-fold among strains (e.g., neuron number ranged from 88,000 to 170,000). Glial and endothelial populations ranged more widely (5- to 8-fold), in part because of higher technical error. A quantitative trait locus (QTL) for the BLAc size is located on chromosome (Chr) 8 near the Large gene. This locus may also influence volume of other regions including hippocampus and cerebellum. Cell populations in the BLAc appear to be modulated more weakly by loci on Chrs 11 and 13. Candidate genes were selected on the basis of correlation with BLAc traits, chromosomal location, single nucleotide polymorphism (SNP) density, and expression patterns in the Allen Brain Atlas. Neurod2, a gene shown to be significant for the formation of the BLAc by knockout studies, is among the candidates genes. Other candidates include Large, and Thra. Responses to drugs of abuse and locomotor activity were the most notable behavioral correlates of the BLAc traits. PMID- 17131201 TI - Aspects of fluency in writing. AB - The notion of 'fluency' is most often associated with spoken-language phenomena such as stuttering. The present article investigates the relevance of considering fluency in writing. The basic argument for raising this question is empirical-it follows from a focus on difficulties in written and spoken language as manifestations of different problems which should be investigated separately on the basis of their symptoms. Key-logging instruments provide new possibilities for the study of writing. The obvious use of this new technology is to study writing as it unfolds in real time, instead of focusing only on aspects of the end product. A more sophisticated application is to exploit the key-logging instrument in order to test basic assumptions of contemporary theories of spelling. The present study is a dictation task involving words and non-words, intended to investigate spelling in nine-year-old pupils with regard to their mastery of the doubling of consonants in Norwegian. In this study, we report on differences with regard to temporal measures between a group of strong writers and a group of poor ones. On the basis of these pupils' writing behavior, the relevance of the concept of 'fluency' in writing is highlighted. The interpretation of the results questions basic assumptions of the cognitive hypothesis about spelling; the article concludes by hypothesizing a different conception of spelling. PMID- 17131202 TI - My mentor, George Albee. PMID- 17131203 TI - George Albee: a man with a better idea. PMID- 17131204 TI - George Wilson Albee (1921-2006), radical community psychologist: a critical obituary. PMID- 17131206 TI - Thrombosed portal vein aneurysm. AB - We describe two cases of thrombosed extrahepatic portal vein aneurysms diagnosed by sonography, computed tomography, and magnetic resonance imaging. Portal vein aneurysm is a rare clinical entity that has been described as a focal dilatation that can affect intra- and extrahepatic portal branches. Although usually asymptomatic, thrombosis can lead to portal hypertension. Clinical and imaging characteristics are discussed, in addition to a review of the literature. PMID- 17131205 TI - Adiponectin and leptin: potential tools in the differential diagnosis of pediatric diabetes? AB - The incidence of type 1 and type 2 diabetes mellitus in the pediatric population has increased over the past decade. The practitioner is often faced with the challenge of differentiating between type 1 and type 2 diabetes at the time of initial diagnosis because of the overlap of clinical and laboratory characteristics between these two entities. Adipokines are proteins secreted by the adipose tissue. Leptin and adiponectin are two adipokines that have been extensively studied in vitro, in animal studies, and in human subjects with type 1 and type 2 diabetes. Leptin and adiponectin play a significant role in the regulation of lipid and carbohydrate metabolism. Adiponectin increases insulin sensitivity in both the liver and skeletal muscle. Leptin decreases appetite, increases energy expenditure, suppresses insulin synthesis and secretion and increases insulin sensitivity. Changes in the secretion or sensitivity to leptin and adiponectin may contribute to the development of type 1 and type 2 diabetes. Adiponectin is higher in adult and pediatric patients with type 1 diabetes compared to those with type 2 diabetes. Data regarding leptin levels are contradictory. Most studies report decreased serum leptin at the time of diagnosis in type 1 diabetes compared to type 2 diabetes subjects and non diabetic controls. This paper will review basic research and clinical evidence supporting the role of adiponectin and leptin in the development of type 1 and type 2 diabetes and discuss their potential use as tools in the differential diagnosis of pediatric diabetes. PMID- 17131207 TI - Metastatic lymph nodes in urogenital cancers: contribution of imaging findings. AB - The presence of nodal metastasis in patients with urogenital malignancies is an important factor for prognosis, and radiologic identification of enlarged nodes greatly affects treatment choices. Radiologic evaluation for nodal metastases is usually performed with computed tomography, but magnetic resonance imaging is also useful in evaluating primary and nodal metastases in pelvic tumors. On these cross-sectional modalities, nodal metastases are usually suspected according to location and size criteria. Although there has been no general consensus on the criteria, a short axis diameter of 8 to 10 mm is generally applied. However, radiologic evaluation does not always provide sufficient accuracy for nodal staging because of an inability to diagnose smaller metastatic lymph nodes. The clinical significance of a radiologic recognition of enlarged nodes also differs by cancer type in relation to differences in staging systems and treatment. The presence of regional lymphadenopathy in patients with renal cell carcinoma often alters surgical methods, whereas the presence of regional lymphadenopathy is an indication of systemic chemotherapy in patients with cancers of the urinary tract, prostate, and testicles. In this report, preferential sites and staging of nodal metastasis and contributions of radiologic imaging are reviewed for each urogenital cancer. PMID- 17131208 TI - Gallbladder adenomyomatosis: imaging findings. AB - In this pictorial essay, we describe the imaging findings of adenomyomatosis of the gallbladder and emphasize high-resolution ultrasound and magnetic resonance cholangiopancreatography in its diagnosis. PMID- 17131209 TI - Percutaneous ultrasound-guided thrombin injection as first-line treatment of pancreatic pseudoaneurysm. AB - Pancreatic pseudoaneurysms are a rare but potentially fatal complication of pancreatitis. Surgical intervention and transcatheter embolization are not always feasible therapeutic options. In this report we present a case of a pseudoaneurysm secondary to pancreatitis which, despite being angiographically invisible, was successfully embolized with a single ultrasound-guided percutaneous injection of thrombin. PMID- 17131210 TI - Conversion of non-tunneled to tunneled hemodialysis catheters. AB - PURPOSE: To determine the safety and efficacy of conversion of non-tunneled (temporary) catheters to tunneled catheters in hemodialysis patients. METHODS: A retrospective review of 112 consecutive conversions in 111 patients was performed over a period of 4 years. Fourteen patients were lost to follow-up. The remaining 97 patients had clinical follow-up. Temporary catheters were converted to tunneled catheters utilizing the same internal jugular venotomy sites and a modified over-the-wire technique with use of a peel-away sheath . Follow-up clinical data were reviewed. RESULTS: Technical success was achieved in all 112 procedures. None of the 97 patients with follow-up suffered early infection within 30 days. The total number of follow-up catheter days was 13,659 (range 2 790). Cases of confirmed and suspected bacteremia requiring catheter removal occurred at a frequency of 0.10 per 100 catheter days. Suspected catheter infection treated with antibiotics but not requiring catheter intervention occurred at a frequency of 0.04 per 100 catheter days. Frequency of all suspected or confirmed infections was 0.14 per 100 catheter days. Catheter interventions as a result of poor blood flow, inadvertent removal, catheter fracture, or kinking occurred at a rate of 0.18 per 100 catheter days. Life table analysis revealed primary patency rates of 86%, 64%, and 39% at 30 days, 90 days, and 180 days, respectively. CONCLUSION: Conversion of temporary catheters to tunneled catheters using the pre-existing venotomy sites is safe and has low rates of infection and malfunction. These rates are comparable to previously published rates for tunneled catheters placed de novo and tunneled catheter exchanges. PMID- 17131211 TI - Application of a new guiding system in percutaneous biopsies. AB - We herein describe the application of a new guiding system designed for percutaneous biopsies. The guiding system set is composed of a 0.41 mm (27G) stainless steel guide stylet and a 22G Chiba needle. Following the initial insertion of the Chiba needle, the stylet is advanced via the needle toward the lesion. The stylet serves either as a guide for the Chiba needle or as an exchange wire for the introduction of larger or cutting biopsy needles. The stylet can also be curved prior to its insertion to facilitate access to lesions which require needle redirection. The technique was applied to 117 cases (54 thoracic, 31 abdominal, 21 pelvic, and 11 vertebral lesions.) The main advantage of the stylet is its small diameter, rendering it atraumatic and permitting multiple punctures for the successful final targeting of the lesion. With this guiding set we achieved targeting of difficult lesions. Furthermore, larger needles were more easily introduced in locations that posed technical difficulties. No major complications were observed. The complication rate was comparable to that of the conventional biopsy technique. The technique using the guide stylet was easily performed and could be applied to almost all organs. PMID- 17131212 TI - Evaluating terrestrial carbon sequestration options for Virginia. AB - Changes in forest and agricultural land management practices have the potential to increase carbon (C) storage by terrestrial systems, thus offsetting C emissions to the atmosphere from energy production. This study assesses that potential for three terrestrial management practices within the state of Virginia, USA: afforestation of marginal agricultural lands; afforestation of riparian agricultural lands; and changing tillage practices for row crops; each was evaluated on a statewide basis and for seven regions within the state. Lands eligible for each practice were identified, and the C storage potential of each practice on those lands was estimated through a modeling procedure that utilized land-resource characteristics represented in Geographic Information System databases. Marginal agricultural lands' afforestation was found to have the greatest potential (1.4 Tg C yr(-1), on average, over the first 20 years) if applied on all eligible lands, followed by riparian afforestation (0.2 Tg C yr( 1) over 20 years) and tillage conversion (0.1 Tg C yr(-1) over 14 years). The regions with the largest potentials are the Ridge and Valley of western Virginia (due to extensive areas of steep, shallow soils) and in the Mid-Atlantic Coastal Plain in eastern Virginia (wet soils). Although widespread and rapid implementation of the three modeled practices could be expected to offset only about 3.4% of Virginia's energy-related CO(2) emissions over the following 20 years (equivalent to about 8.5% of a Kyoto Treaty-based target), they could contribute to achievement of C-management goals if implemented along with other mitigation measures. PMID- 17131213 TI - Perceptual adaptation to spectrally shifted vowels: training with nonlexical labels. AB - Although normal-hearing (NH) and cochlear implant (CI) listeners are able to adapt to spectrally shifted speech to some degree, auditory training has been shown to provide more complete and/or accelerated adaptation. However, it is unclear whether listeners use auditory and visual feedback to improve discrimination of speech stimuli, or to learn the identity of speech stimuli. The present study investigated the effects of training with lexical and nonlexical labels on NH listeners' perceptual adaptation to spectrally degraded and spectrally shifted vowels. An eight-channel sine wave vocoder was used to simulate CI speech processing. Two degrees of spectral shift (moderate and severe shift) were studied with three training paradigms, including training with lexical labels (i.e., "hayed," "had," "who'd," etc.), training with nonlexical labels (i.e., randomly assigned letters "f," "b," "g," etc.), and repeated testing with lexical labels (i.e., "test-only" paradigm without feedback). All training and testing was conducted over 5 consecutive days, with two to four training exercises per day. Results showed that with the test-only paradigm, lexically labeled vowel recognition significantly improved for moderately shifted vowels; however, there was no significant improvement for severely shifted vowels. Training with nonlexical labels significantly improved the recognition of nonlexically labeled vowels for both shift conditions; however, this improvement failed to generalize to lexically labeled vowel recognition with severely shifted vowels. Training with lexical labels significantly improved lexically labeled vowel recognition with severely shifted vowels. These results suggest that storage and retrieval of speech patterns in the central nervous system is somewhat robust to tonotopic distortion and spectral degradation. Although training with nonlexical labels may improve discrimination of spectrally distorted peripheral patterns, lexically meaningful feedback is needed to identify these peripheral patterns. The results also suggest that training with lexically meaningful feedback may be beneficial to CI users, especially patients with shallow electrode insertion depths. PMID- 17131214 TI - Comorbid substance abuse and neurocognitive function in recent-onset schizophrenia. AB - Despite the high prevalence of comorbid substance use disorder (SUD) up to 65% in schizophrenia there is still few knowledge about the influence of substance abuse on neurocognitive function. In a prospective design we recruited 68 patients (aged 18-40 years) diagnosed as recent-onset schizophrenia or schizoaffective disorder consecutively admitted to hospital. The patients received standardized psychopathological evaluation of schizophrenic symptoms [Brief Psychiatric Rating Scale (BPRS), Scale for the Assessment of Negative Symptoms (SANS)], depressive symptoms [Montgomery Asberg Depression Rating Scale, (MADRS)] and global ratings [Clinical Global Impressions Scale (CGI), Global Assessment of Functioning Scale (GAF)]. Out of this sample 44 subjects underwent after stabilization (4-6 weeks after admission) neuropsychological investigation focusing on early information processing (Trail-Making-Test A, Digit Span), visuo-spatial ability (Corsi Block Tapping), verbal fluency (Verbal Fluency Test, semantic and letter category), and executive functioning and cognitive flexibility [Trail-Making-Test B, Wisconsin Card Sorting Test (WCST)]. About 36% of patients reported drug abuse [European Addiction Severity Index (EuropASI)] with a high prevalence for cannabis. Compared with nonabusers the sample of substance abusers was younger, predominantly male and had lower socioeconomic status. Attentional impairment according to the SANS subscale was less in abusers than in nonabusers on admission, no other psychopathological differences could be detected. Schizophrenic patients without substance abuse demonstrated significantly better performance only in a few neurocognitive tasks (Verbal Fluency, letter category and at trend level Digit Span, backwards), while there tended to be an advantage for substance abusers in executive functioning (WCST, not significant). These results are consistent with other studies of first-episode patients. The lack of higher cognitive disturbance in young schizophrenic patients with comorbid substance abuse may encourage clinicians to develop integrated treatment programmes using cognitive strategies of drug therapy. PMID- 17131215 TI - Sleep quality in adult patients with attention deficit hyperactivity disorder (ADHD). AB - Sleep disorders are common in adults with attention deficit/hyperactivity disorder (ADHD). Two sleep questionnaires; the Brown ADD Scale, the Symptom Checklist-90-R, and a self-developed symptom questionnaire were administered in this study. In a sample of 61 patients without comorbidity, current substance abuse and medication intake, sleep problems were reported more often than in healthy controls. A lack of being refreshed in the morning was very closely associated with ADHD symptomatology whereas insomnia was related to the presence of comorbidity and depressive symptoms. In the total sample (N = 120), medication intake (mainly stimulants) was not related to any of the sleep parameters. It seems important to screen adult patients with ADHD for the presence of sleep disorders, especially insomnia, restless legs syndrome, and sleep-related breathing disorders (the last in conjunction with the body mass index). Since primary sleep disorders are associated with cognitive impairment, one might expect that ADHD symptomatology may improve if comorbid sleep disorders are adequately treated in addition to the specific ADHD treatment. PMID- 17131216 TI - The spectra of neurasthenia and depression: course, stability and transitions. AB - BACKGROUND: Neurasthenia has had a chequered history, receiving changing labels such as chronic fatigue or Gulf war syndrome. Neurasthenia is recognized by ICD 10, but not by DSM-IV. Its course, longitudinal stability and relationship to depression is not well understood. METHODS: In a stratified community sample (n = 591), representative of 2600 persons of the canton of Zurich, Switzerland, neurasthenia and depression were assessed in six structured interviews between ages 20 and 41. Course, stability and comorbidity were examined. A severity spectrum of neurasthenia and depression from symptoms to diagnosis was taken into account. RESULTS: The annual prevalence of a neurasthenia diagnosis increased from 0.7% to 3.8% from age 22-41, while mere symptoms became less prevalent. Intraindividual courses improved in 40% and deteriorated in about 30% of symptomatic cases. The most frequent symptoms overall, besides criterial exhaustion, were increased need for sleep, over-sensitivity, nervousness and difficulty concentrating. Cross-sectional associations and overlap with depression were strong. Longitudinal stability of ICD-neurasthenia was low. CONCLUSIONS: Neurasthenia is intermittent, overlaps significantly with depression, and shows improvement and deterioration over time to roughly equal measures. PMID- 17131217 TI - Post-stroke affective or apathetic depression and lesion location: left frontal lobe and bilateral basal ganglia. AB - This study was designed to examine the correlation between damage to the basal ganglia or frontal lobe and depression status (both affective and apathetic dimensions) in 243 stroke patients. We assessed the affective dimension in post stroke depression (PSD) using the Zung Self-rating Depression Scale (SDS) and the apathetic dimension in PSD using the apathy scale (AS). We classified basal ganglia or frontal lobe damage into four groups: no damage, damage to the left side only, damage to the right side only, and damage to both sides. Affective and/or apathetic PSD was found in 126 patients (51.9%). The severity of affective depression (SDS score) was associated with left frontal lobe (but not basal ganglia) damage, and that of apathetic depression (AS score) was related to damage to the bilateral basal ganglia (but not to the frontal lobe). The anatomical correlates of PSD differ depending on the PSD dimension (affective or apathetic) and may explain interstudy differences regarding the association between lesion location and type of PSD. PMID- 17131218 TI - The current state of adult mental health care in France. AB - Since the 1960s, French public mental health services are organised in "sectors", each sector catering for a mean population of 54,000 inhabitants. Although this organisation was aimed at insuring equal access to care whatever the place of residence, there are still huge disparities in number of staff and bed resources from one sector to another. The reduction in the number of hospital beds started later in France than in most other European countries, and was really effective in the 1990s. In 2000, there were 9.4 beds for 10,000 inhabitants aged over 20 years. Hospital-based care has still an overwhelming importance, and is associated with a marked under-development of community services and lack of sheltered housing for the most disabled patients. With more than 13,000 registered psychiatrists in France, the density of psychiatrists is one of the highest in the world. However, French psychiatry has currently to face a structural crisis due to the reduction in public health budgets, as well as to the reduction of 30% in the number of French psychiatrists over the next two decades. The numerous national programmes aimed at renovating French mental health services, published over the last decade, have not yet kept their promises. PMID- 17131219 TI - Gonadal axis hormones in psychiatric male patients after a suicide attempt. AB - Epidemiological and clinical studies support the view that aggressive acts like suicidal and violent behaviors share a common substrate. Certain aspects of violence in males have been related to high testosterone levels, but the relation of testosterone to attempted suicide has not been studied until now. We estimated plasma levels of testosterone (T), LH, and FSH in 80 male subjects after a suicide attempt and in whom a psychiatric assessment was done during their hospitalization. Suicide intent was evaluated in 72 subjects. A group of 56 healthy males in the same age range served as control. As a group, attempters showed significantly lower T levels, marginally higher LH, and normal FSH compared to controls. The attempters who used violent methods (26 subjects) had T levels even lower than the non-violent (drug overdose) subgroup. Comparisons of T levels of subgroups according to the (main) drug ingested (analgesics, benzodiazepines, antidepressants, neuroleptics, or other drugs) did not reveal any significant drug effect. In relation to diagnosis, the lowest T levels were found in the subgroup with schizophrenia (29 subjects). The T levels of this subgroup were also significantly lower compared to those of a group of 31 male schizophrenic patients, hospitalized and treated with neuroleptics. If the influence of post-attempt stress and medical condition on plasma T could be ruled out, low plasma T may prove to be a biological predictor of attempt, at least in male schizophrenic patients. Nevertheless, the findings differentiate suicidal behavior from other aggressive/violent behaviors and do not support the notion that suicidal and aggressive behaviors are manifestations of the same impulse. PMID- 17131221 TI - Camptocormia in Parkinson's disease. AB - Camptocormia is defined as an abnormal, severe and involuntary forward flexion of the thoracolumbar spine, which becomes manifest during standing and walking and subsides in the recumbent position. It was originally described as a psychogenic disorder, particularly in soldiers involved in long-term trench service during World War 1. It is becoming increasingly recognized as a prominent and disabling phenomenon during the course of Parkinson's disease (PD). In our experience, there is no clear correlation between camptocormia and levodopa treatment. In a few patients, the abnormal posture improved and in others it was unaltered or even became worse following levodopa administration. In a minority of fluctuating patients, there was a temporary deterioration during the "off" periods, but in most, the severity of camptocormia was unchanged during the "on" and "off" phases. In some patients it is associated with back pains, whereas in others it is painless. It occurs in sporadic PD as well as in postencephalitic and parkin gene mutation PD and in other parkinsonian syndromes such as MSA. The pathogenesis of this striking clinical sign is unknown. It is definitely not due to a primary vertebral disease causing kyphosis such as ankylosing spondylitis, as the bent spine disappears when the patient lies on his back. The muscles involved may be the abdominal, paravertebral or both. It may by due to a peculiar dystonia or to an extreme form of rigidity. Local myopathic changes were suggested as a possible cause, but these may rather be a secondary phenomenon. Treatment is currently unsatisfactory in most cases. Occasional patients may benefit from intramuscular botulinum toxin injections or from deep brain stimulation. PMID- 17131222 TI - Lateral flexion in Parkinson's disease and Pisa syndrome. AB - Various types of abnormal posture are observed in Parkinson's disease (PD). Lateral flexion is very common and frequent among them. The clinical characteristics of lateral flexion in PD vary and are classified into two types, the chronic and subchronic types. The chronic type of lateral flexion in PD appears subclinically and worsens, which is related to the laterality of parkinsonian symptoms and the progression of the disease. The subchronic type of lateral flexion in PD develops subacutely and worsens rapidly in several months. An atypical and rare type of tonic truncal dystonia, Pisa syndrome, may be induced following the intake of neuroleptics. The clinical features of the subchronic type of lateral flexion in PD are similar to those of Pisa syndrome. Differences between lateral flexion in PD and Pisa syndrome are described. PMID- 17131223 TI - The natural history of Parkinson's disease. AB - Although Parkinson's disease (PD) is the only chronic neurodegenerative disorder for which there are effective symptomatic therapies no treatment has yet been identified that would significantly slow its natural progression. Studies into the natural history of Parkinson's disease have been complicated by issues of diagnostic accuracy, heterogeneity of different forms of the disease as well as confounding effects of age related comorbidities. Only recently results from placebo-controlled clinical trials have helped to define the rates of progression of motor dysfunction as assessed by the UPDRS in early PD. Studies in treated patients suggest that there may be different rates of progression in different phases of the disease with faster decline in earlier versus later stages. Measuring progression of motor symptoms, however, alone is insufficient to describe the natural history of PD and may also be inadequate to define clinically meaningful disease modification in intervention trials. Progression of global disability as captured in the Hoehn and Yahr scale is important to consider and progression to stage III with beginning postural impairment defines a major milestone in the natural history of this illness. In addition, non-motor symptoms have major impact on the natural history of PD. Dementia, sleep-wake cycle dysregulation and autonomic failure were present in 50-80% of patients in one recent 15 year-follow up study. Dementia and psychosis are also major risk factors for nursing home placement in PD. A prerequisite for effective interventions that would modify disease progression in Parkinson's disease is the identification of the earliest preclinical stages of illness. This is now become a realistic possibility with screening tests of olfactory function and subsequent functional imaging. Subjects at risk for Parkinson's disease should be the future focus of neuroprotective trials. PMID- 17131224 TI - Dropped head in Parkinson's disease. AB - "A propensity to bend the trunk forward" and "the chin is now almost immovably bent down upon the sternum" were described by James Parkinson in patients with Parkinson's disease (PD). The term "dropped head" was first reported in "Gerlier disease" in Switzerland and 'kubisagari' in Japan and since then also reported in myositis, myopathy, myasthenia gravis, amyotrophic lateral sclerosis, neuropathy, and hypothyroidism. Disproportionate antecollis occurs in about half cases of multiple system atrophy (MSA) and is considered dystonic in nature. Dropped head is considered rare in PD, both in advanced and early stages of PD. However, it is known to progress subacutely over a period of several days. In my experience, dropped head is relatively common in PD. The mechanism of dropped head in PD is either dystonia of flexor neck muscles or weakness of extensor neck muscles. The response of dropped head to various anti-parkinsonian medications is rather inconsistent. Levodopa is reported to induce amelioration in some patients while dopamine agonists can cause deterioration. Muscle afferent block with lidocaine and ethanol is reported to be effective, while the effect of botulinum toxin injection into the affected muscles is limited. The effect of stereotaxic neurosurgery on dropped head is controversial. Early diagnosis and prompt treatment is necessary to prevent muscle damage associated with longterm overstretch of extensor neck muscles. PMID- 17131225 TI - Freezing of gait in Parkinson's disease. AB - Freezing of Gait (FOG) is one of the most disabling and least understood symptoms in Parkinson's disease (PD), and is usually observed in the advanced stage of the disease. FOG can be experienced on turning, in narrow spaces, whilst reaching a destination, and in stressful situations. FOG is commonly observed in the "off" state, but it can also be observed in the "on" state. Dual tasking (cognitive load) aggravates FOG. Visual or auditory cues often resolve FOG. Analysis of gait revealed that the rhythm of stepping suddenly jumps into high frequency (4-5 Hz) in FOG (hastening), and that floor reaction forces are disregulated. Stride-to stride variability is increased in FOG. Hastening phenomenon was reported not only in PD patients but also in patients with striatal lesions. The basal ganglia and its frontal projections may be one of the essential lesion sites for FOG.A recent study using single-photon emission tomography (SPECT) revealed enhanced lateral premotor cortex (PMC) activity during paradoxical gait in PD, suggesting that PMC can compensate for the impaired function of the medial frontal cortex when cued by visual input. Treatment of FOG includes behavioural, medical, and surgical approaches. Tricks of all kinds (including external cues) are effective therapeutic approaches. If FOG occurs predominantly in the "off" state, dopaminergic therapy can be increased. For "on" freezing or if "on" response is otherwise optimised, the dose of the dopaminergic agent may be manipulated, but it could lead to the deterioration of parkinsonism. Deep brain stimulation of the STN often alleviates FOG in the "off" state. PMID- 17131226 TI - Restless legs syndrome and Parkinson's disease: is there an etiologic link? AB - The etiologic link between restless legs syndrome (RLS) and Parkinson's disease (PD) has been debated. Since dopaminergic dysfunction and response to dopaminergic agents are consistent features in RLS and PD, some authors have suggested that these two diseases may share common pathophysiology. However, presently there is not enough evidence to suggest that the actual pathophysiologic mechanism in both diseases is identical. The nigrostriatal dopaminergic system is primarily involved in PD and it is possible that the extrastriatal dopaminergic system may be variably involved in those PD patients with RLS symptoms. Further clinical, imaging, pharmacologic, and genetic studies will be needed to address the many unanswered questions related to the link between RLS and PD. PMID- 17131227 TI - Weight loss in Parkinson's disease. AB - Weight loss is frequent in patients with Parkinson's disease (PD). Reduced energy intake and/or increased energy expenditure have been postulated as the cause. Dysphagia, anorexia, and gastrointestinal dysfunction may be possible causes of reduced energy intake; whereas, rigidity, tremor, and levodopa-induced dyskinesia may increase energy expenditure. Levodopa may enhance glucose metabolism resulting in enhanced energy expenditure. Depression, anti-parkinsonian drugs, and medical complications such as pneumonia and malignancies also may facilitate weight loss in PD. Combinations of various degrees of these factors, especially in advanced PD, may produce weight loss. Such weight loss is associated with malnutrition which may precipitate infection and decubitis; accelerate motor, behavioral, and autonomic impairment; consequently spoiling one's quality of life. Attention must be paid as well to motor symptoms to prevent or reverse weight loss in PD patients. PMID- 17131228 TI - Sweating dysfunctions in Parkinson's disease. AB - There is a high frequency of dyshidrosis in Parkinson's disease. Daily use of an antiparkinsonian drug does not affect sweating. Mental sweating relates to the contraction period and seriousness. However, hyperthermic sweating does not necessarily relate to the contraction period or seriousness. Abnormalities in mental sweating are not necessarily correlated with cardiovascular autonomic disturbances. As the autonomic disturbance becomes more advanced, dyshidrosis becomes more common. Hyperhidrosis may develop with dyskinesia. Hyperhidrosis may be improved by the temporally administration of levodopa. Dyshidrosis might be caused by a centrally-acting abnormality during its early stage. However, postganglional abnormalities as well as central changes may increase as the disease progresses. The presence of dyshidrosis affects the QOL and depression in patients with Parkinson's disease. The only therapy for hyperhydrotic sweating disorders is the administration of levodopa or dopaminagonist. Additional studies including therapy for sweating disorders are necessary. PMID- 17131229 TI - Parkinson's disease and fatigue. AB - Fatigue is one of the most common symptoms in patients with Parkinson's disease (PD), and its impact on the quality of life is substantial. However, its cause and treatment are not established. Fatigue in PD has two components, peripheral and central, which may be related to each other, but are more likely independent. Fatigue is partially associated with depression or sleep disorders, but patients with fatigue are not always depressed and do not necessarily have sleep problems. Anti-PD drugs may exacerbate or reduce fatigue. The impact of fatigue in PD is often underestimated by health-care providers. PMID- 17131230 TI - Apathy and the basal ganglia. AB - We should like to emphasize the following points: 1. Apathy is defined here as a quantified and observable behavioral syndrome consisting in a quantitative reduction of voluntary (or goal-directed) behaviors; 2. Therefore, apathy occurs when the systems that generate and control voluntary actions are altered; 3. These systems are mostly represented by the different subregions embedded in the Prefrontal cortex (PFC) and in the basal ganglia regions that are closely connected with the PFC; 4. In consequence, clinically, apathy is a prefrontal syndrome either due to direct lesions of the PFC or to lesions of basal ganglia areas that are closely related to the PFC; 5. Apathy is not a single entity but rather heterogeneous. Several different mechanisms may lead to apathy; Because there are several anatomical-functional prefrontal-basal ganglia circuits, the underlying mechanisms responsible for apathy may differ according to which prefrontal-basal ganglia circuit is affected; 6. In this context, apathy is the macroscopic results of the disruption of one or several elementary steps necessary for goal-directed behavior that are subserved by different prefrontal basal ganglia circuits; 7. Intense apathy is related to caudate nucleus and GPi, disrupting associative and limbic pathways from/to the PFC; 8. in progressive supranuclear palsy (PSP) and focal lesions (caudate nuclei, GPi), apathy may be due to a loss of PFC activation; 9. In Parkinson's disease (PD), apathy may be due to a loss of signal focalization; 10. More globally, we propose that apathy may be explained by the impact of lesions or dysfunctions of the BG, because these lesions or dysfunctions lead to a loss of amplification of the relevant signal and/or to a loss of temporal and spatial focalization, both of which result in a diminished extraction of the relevant signal within the frontal cortex, thereby inhibiting the capacity of the frontal cortex to select, initiate, maintain and shift programs of action. PMID- 17131231 TI - Dystonia in Parkinson's disease. AB - Dystonia can occasionally be found in idiopathic Parkinson's disease. It is very uncommon in untreated patients and is more frequently seen as a complication of its treatment. In this review, the various types of dystonia occurring in PD, the differential diagnosis with other parkinsonian syndromes associated with dystonia and treatments available are revised. Dystonia unrelated to treatment can be typical (blepharospasm, torticollis), atypical (parkinsonian writer's cramp, camptocormia, anismus), or occurring in earlyonset Parkinson disease (the so called kinesigenic foot dystonia, considered a hallmark of early-onset Parkinson's disease). Early and prominent dystonia in untreated patients with parkinsonism should raise the suspicion of other entities other than Parkinson's disease, such as progressive supranuclear palsy, multiple system atrophy or corticobasal degeneration. In patients on chronic dopaminergic treatment, peak dose dystonia, diphasic dystonia and off-dystonia can be seen. The later constitutes the major dystonic feature of chronic levodopa therapy, and a wide variety of strategies are available to manage this complication. Among them, deep brain stimulation of the subthalamic nucleus has proved to be the most effective one. Dystonic reactions (mainly involving oculomotor cranial nerves and limbs) in operated patients (especially carriers of deep brain stimulation (DBS) devices) are increasingly being reported, constituting a new type of dystonia in patients with Parkinson's disease: dystonia linked to surgical treatment. PMID- 17131232 TI - The role of phospholipase A2 in neuronal homeostasis and memory formation: implications for the pathogenesis of Alzheimer's disease. AB - Phospholipase A(2) (PLA(2)) is a key enzyme in cerebral phospholipid metabolism. Preliminary post-mortem studies have shown that PLA(2) activity is decreased in frontal and parietal areas of the AD brain, which is in accordance with recent (31)P-Magnetic Resonance Spectroscopy evidence of reduced phospholipid turnover in the pre-frontal cortex of moderately demented AD patients. Such abnormality may also be observed in peripheral cells, and reduced PLA(2) activity in platelet membranes of AD patients, and correlates with the severity of dementia. In rat hippocampal slices, PLA(2) has been implicated in mechanisms of synaptic plasticity. In adult rats, the stereotaxic injection of PLA(2) inhibitors in the CA1 area of hippocampus impaired, in a dose-dependent manner, the formation of short- and long-term memory. Additionally, such inhibition resulted in a reduction of the fluidity of hippocampal membranes. In primary cultures of cortical and hippocampal neurons, the inhibition of PLA(2) precluded neurite outgrowth, and the sustained inhibition of the enzyme in mature cultures lead to loss of viability. Taken together, these findings reinforce the involvement of PLA(2) enzymes in neurodevelopment and neurodegeneration processes, and further suggest that reduced PLA(2) activity, probably reducing membrane phospholipids breakdown, may contribute to the memory impairment in AD. PMID- 17131233 TI - Proceedings of the 39th Meeting of the Austrian Society of Tropical Medicine and Parasitology, 17-19 November 2005, Vienna, Austria. PMID- 17131234 TI - Vaccines against traveler's diarrhoea and rotavirus disease - a review. AB - Diarrheal diseases constitute one of the most important health problems worldwide, preferentially in developing countries with a morbidity of estimated 5 billion and a mortality of 5 million cases per year. Children less than 5 years are particularly in danger with respect to the incidence and severity of the gastrointestinal symptoms. Travelers to developing countries are also at risk to develop diarrheal disorders; around 30-50% of them acquire so called "travelers's diarrhea" caused by bacteria, viruses or protozoa. It has been estimated that approximately 30-70% of diarrhea are due to bacteria, of which the most frequently detected enteric pathogens are non-invasive, enterotoxigenic Escherichia coli (ETEC). Their exotoxins, the heat stabile (ST) and the heat labile (LT) toxins are in large part responsible for the pathogenicity of the bacteria. About 20% of cases of traveler's diarrhea are caused by LT producing ETEC. This heat labile toxin exhibits a 80% sequence homology with cholera toxin. The presently available vaccine against cholera (Dukoral) contains inactivated Vibrio cholerae bacteria and the recombinant non-toxic B subunit of cholera toxin. Consequently, this vaccine displays also some efficacy against traveler's diarrhoea with up to 25% of travelers being protected against this disease. Rotaviruses are the leading recognized cause of diarrhoea-related illness and deaths among infants worldwide in developing and industrialized countries. Based on the high incidence of this disease two oral vaccines have been developed and will be available in Europe in 2006. Due to the impact of rotavirus diseases also in Austria vaccination against this disease has been already suggested in the Austrian vaccination schedules for infants from 6-24 weeks of age. One of the two vaccines, Rotarix, is an attenuated monovalent vaccine with a broad cross reactivity against the most frequent serotypes. The second one, RotaTeq, is a pentavalent attenuated vaccine containing 5 human-bovine reassortants. Both vaccines display 85-98% efficacy against severe rotavirus disease and an excellent tolerability with no difference in side reactions to the placebo controls, particularly with respect to intussusceptions. With respect to increasing travel habits with infants and small children, particularly when visiting friends and relatives, vaccination against rotavirus infections will also be important in international travel. PMID- 17131235 TI - Genetic vaccination approaches against malaria based on the circumsporozoite protein. AB - Malaria is the world's major parasitic disease, for which effective control measures are urgently needed. Despite considerable efforts, no successful vaccine against malaria has been developed so far. The method of DNA-based immunization offers the possibility to induce both antibody- and cell-mediated immune responses to a variety of antigens. The flexibility of the DNA vaccine technology permits the combination of several antigens from different developmental stages of the parasite's complicated life cycle. This review covers the development of DNA-based immunization against malaria from initial experiments in small animals to recently conducted clinical studies. Focusing on one of the best characterized malaria vaccine candidate antigens, the circumsporozoite protein, an overview of strategies to enhance vaccine efficacy is provided. Advanced application methods such as the gene gun technology or the needle-less jet injection device are described. As DNA vaccination represents a relatively new methodology, safety concerns associated with planned clinical applications are discussed. In summary, this novel type of vaccine has to be considered as a promising tool for future malaria vaccination strategies. PMID- 17131236 TI - [Relevance of parasitological examinations for the clinical course, epidemiology and prevention of alveolar echinococcosis - experiences of more than two decades in Austria]. AB - Alveolar echinococcosis has been known in Austria since 1854. At the beginning of the 20th century the Austrian researcher on echinococcosis, Adolf Posselt, has realized already the enormous medical relevance of serological tests for the diagnosis of alveolar echinococcosis as well as for preventive purposes. Since the beginning of the 1980ies a long-term study about the epidemiology and serodiagnosis of alveolar echinococcosis has started in the Department of Medical Parasitology of the Clinical Institute of Hygiene and Medical Microbiology of the University of Vienna. Today a highly sensitive enzyme-linked immunosorbent assay as well as a highly specific westernblot system are available for the serological diagnosis of human alveolar echinococcosis cases, for the serological monitoring of the disease as well as for seroepidemiological and preventive medical studies. In addition, molecular biological tools for the detection of E. multilocularis (as well as E. granulosus strain) specific DNA in surgical resected and bioptic materials complete our diagnostic spectrum of methods. PMID- 17131237 TI - [Mites as a cause of zoonoses in human beings]. AB - Different mite species occurring in animals may infest humans temporarily. Such agents should be considered a possible cause of erythemateous and sometimes pruritic skin reactions of unclear origin. Pseudoscabies is a common problem in occupationally exposed humans, e.g. farmers, veterinarians or pet owners. Those selflimiting dermatoses may often be misdiagnosed. Several species including Sarcoptes scabiei, Notoedres cati, Cheyletiella spp., Dermanyssus gallinae, Ornithonyssus bacoti, Ophionyssus natricis and Neotrombicula autumnalis may infest human skin, causing symptoms. PMID- 17131238 TI - [An introduction into bioethics and parasitology]. AB - Bioethical considerations increasingly affect the process of integrating scientific and medical findings into the moral conceivabilities of a society and, consequentially, into the stock of laws. In a natural science like parasitology a social consensus must be reached in the areas of species protection, of the red biotechnology, of human prenatal diagnosis, and of animal experiments. The crucial problem is the social value of an individual's dignity and the conjoined question of the admissibility of an exploitation of a creature. The responsibility of man for other organisms, in that particular case parasites and their hosts, is brought up for discussion in a bioethic debate. PMID- 17131239 TI - Comparison of the proteome profiles of Entamoeba histolytica and its close but non-pathogenic relative Entamoeba dispar. AB - The microaerophilic protist Entamoeba histolytica is a source of considerable morbidity and mortality in many countries of the world by acting as a causative agent of amoebic dysentery and liver abscess. Its close relative, Entamoeba dispar, also colonises the human colon but is non-pathogenic. In the present study, we wanted to assess if the close relatedness of these two organisms could be used to identify virulence factors in E. histolytica through a comparison of the protein expression profiles of the pathogenic and the non-pathogenic species. We applied two-dimensional polyacrylamide gel electrophoresis in order to identify proteins which are specifically expressed in E. histolytica and which therefore could constitute candidate proteins potentially involved in E. histolytica pathogenicity. The evaluation of overall protein expression profiles derived from whole cell extracts, however, revealed that, in spite of the close relatedness at the genetic level, the disparity of the proteomic profiles of these two Entamoeba species is far too great in order to pinpoint proteins whose expression might render E. histolytica pathogenic. Taken together, our study performed at the proteomic level clearly supports the notion of E. histolytica and E. dispar as two distinct species. PMID- 17131240 TI - Development of a pharmacodynamic screening model with Crithidia fasciculata. AB - The genus Crithidia is a member of the family Trypanosomatidae and is related to the genera Leishmania and Trypanosoma with which it shares a variety of biochemical mechanisms, such as polyamine synthesis and methionin salvage. In consequence, a screening system for antiparasitic candidate material has been developed with Crithidia fasciculata, a parasite naturally occurring in insects and amphibians, but devoid of pathogenicity for humans. Initially a variety of culture media were evaluated of which TPS was best suited for the maintenance of stock cultures, and E-medium - a newly developed formula - for sensitivity testing. Optimal growth of C. fasciculata was observed under microaerophilic conditions. A system for sensitivity testing was developed and applied to the investigation of extracts from higher tropical plants of the genera Stemona and Aglaia for anticrithidial activity. Extracts with significant anti-crithidial activity were scheduled for chromatographic fractionation and the subsequent isolation, purification and structural identification of individual compounds for further sensitivity testing. Encouraging results were obtained with extracts from Aglaia odorata leaves, A. elaeagnoidea stem bark and A. edulis leaves, with EC(90) values of 1213 ng/ml, 1606 ng/ml, and 1462 ng/ml, respectively. PMID- 17131241 TI - Optimized Malaria-antigens delivered by immunostimulating reconstituted influenza virosomes. AB - Malaria remains a serious cause of morbidity and mortality in millions of individuals each year. The development of widespread resistance of the parasite to drugs as well as resistance of the transmitting mosquito-vector to insecticides in combination with the poor economic situation of many malaria endemic countries make the development of an effective and inexpensive treatment and prevention a main focus of research. Vaccines remain to be one of the most cost effective and feasible means of disease control and have remarkable success in the control of many infectious disease: eradication of small pox, virtual eradication of polio and the reduction of measles and diphtheria. Next generation vaccines should focus on specific antigens rather than whole inactivated or attenuated pathogens, since the requirements by regulatory authorities concerning safety are becoming more stringent over time. But sub-unit and in particular peptide-based vaccines are poorly immunogenic themselves, and alum represents only a sub-optimal adjuvant for recombinant proteins and synthetic peptides. This emphasizes the need for suitable carrier- and adjuvant systems promoting protective immune responses by delivering protein and peptide antigens in an appropriate conformation. Here, we review the development of a new approach combining peptide-based malaria vaccine candidate antigens with an immune stimulatory carrier-system based on influenza virosomes focusing on the induction of protective antibodies. PMID- 17131242 TI - [In vitro antimalarial drug resistance in Southeastern Bangladesh]. AB - Particularly in Southeast Asia drug resistance has become a major constraint in the treatment of falciparum malaria. So far relatively little is known about the current status of drug resistance in Bangladesh. The aim of this study was therefore to determine the in vitro drug susceptibility of Plasmodium falciparum in south-eastern Bangladesh. In the HRP2 in vitro drug sensitivity assay the tested isolates demonstrated a relatively high sensitivity to dihydroartemisinine (IC50 = 1.33 nM; 95% CI: 1.08-1.63; IC90 = 2.65 nM; 95% CI: 2.13-3.29), mefloquine (IC50 = 11.26 nM, 95% CI: 9.75-13.0; IC90 = 19.55 nM, 95% CI: 15.73 24.29) and quinine (IC50 = 73.24 nM, 95% CI: 65.26-82.21; IC90 = 157.75 nM, (95% CI: 134.16-185.5) thus being significantly more sensitive to mefloquine and quinine than isolates from Thailand. Chloroquine (IC50 = 93.06 nM, 95% CI: 80.38 107.76; IC90 = 214.76 nM, 95% CI: 175.64-262.62) sensitivity was highly compromised with inhibitory concentrations reaching levels comparable to Thailand. Therefore this drug should not be used in the treatment of falciparum malaria in this region. Despite compromised in vitro drug sensitivity to sulfadoxine/pyrimethamine, in clinical studies the combination of sulfadoxine (IC50 = 40.46 microM, 95% CI: 31.15-51.97; IC90 = 173.48 microM, 95% CI: 120.78 249.17) and pyrimethamine (IC50 = 1.7 microM, 95% CI: 1.25-2.3; IC90 = 4.83 microM, 95% CI: 3.17-7.37) with quinine proved to be an interesting option for treating uncomplicated falciparum malaria in Bangladesh. PMID- 17131243 TI - In vitro interaction between artemisinin and chloroquine as well as desbutyl benflumetol in Plasmodium vivax. AB - Malaria resulting from infection with Plasmodium vivax rarely causes death, however, patients usually suffer acute debilitating clinical symptoms and the recovery is slow. This study had the objective of assessing the pharmacodynamic interaction between artimisinin and chloroquine with a view of a potential acceleration of the clinicalparasitological response, and the investigation of therapeutic alternatives in the event of chloroquine resistance in Plasmodium vivax. Tests were based on the growth inhibition of Plasmodium vivax, determined by morphological differential counts of 200 asexual parasites. In total 45 isolates were evaluated successfully with parallel tests for artemisinin, chloroquine and desbutylbenflumetol (DBB) alone and combinations of artemisinin + chloroquine and artemisinin + DBB. Total inhibition was reached at a mean concentration of 1274.8 nM (95% CI 898.5 to 1808.7 nM), and 1852.2 nM (95% CI 1539.5 to 2228.6 nM) for artemisinin, and chloroquine respectively, whilst the 1:1 (m/m) combination of artemisinin and chloroquine was 1860.2 nM (95% CI 1454.4 to 2379.3 nM). EC(50) and EC(90) were 129.9 nM and 1058.5 nM for chloroquine, 32.6 nM and 735.5 nM for artemisinin, and 73.6 nM and 1103.0 nM for the 1:1 combination of both drugs. Interaction analysis according to Berenbaum yielded for the artemisinin + chloroquine combination at the EC(50) a mean SigmaFIC of 1.1126, at the EC(90) a mean SigmaFIC of 1.0331, and at the EC(99) a mean SigmaFIC of 1.1857. These results revealed marked additive interaction. For desbutylbenflumetol (DBB) the EC(50) and EC(90) were 1.5 nM and 28.8 nM, complete growth inhibition was observed at 90.4 nM (95% CI 75.1 to 108.7 nM). Interaction analysis indicated moderate antagonism at the lower concentration ranges, at the EC(90) additive interaction with a mean SigmaFIC of 1.0300, and synergism at the therapeutically most important EC(99) with a mean SigmaFIC of 0.5990. PMID- 17131244 TI - Pharmacodynamic interaction between atovaquone and other antimalarial compounds against Plasmodium falciparum in vitro. AB - Atovaquone, a 2-hydroxy-1,4-naphthoquinone, was first introduced as a drug against opportunistic infections in immuno-compromised patients. Early clinical parasitological experiences in the treatment of malaria were disappointing due to highly variable and poor absorption, a phenomenon typical for naphthoquinones. Proguanil was found to potentiate the activity of atovaquone and the combination of the two drugs was introduced as an antimalarial drug with blood schizontocidal and causal prophylactic activity. It is now widely used in therapy and prophylaxis. Despite the enhanced activity, the combination does not always overcome the problem of poor absorption of atovaquone, especially in the presence of gastro-intestinal disorders. Therefore, further combination with a fast-acting blood schizontocide, e.g. one of the artemisinins, could accelerate clinical improvement and normalization of absorption. The interaction between artemisinin and atovaquone and that of artemisinin and atovaquone + proguanil has been investigated in 37 fresh isolates of Plasmodium falciparum from northwestern Thailand, an area with high prevalence of multi-drug resistance. Interaction between atovaquone and artemisinin was synergistic above the EC(30), with mean SigmaFIC (Berenbaum) values of 0.9679 at the EC(50), 0.4014 at the EC(90) and 0.2214 at the EC(99). Synergism was more pronounced with the triple combination, i.e. atovaquone + proguanil and artemisinin, starting at the EC(10) level. The mean SigmaFIC values were 0.7626 at the EC(50), 0.2939 at the EC(90), and 0.1527 at the EC(99). The strong synergism at the therapeutically relevant effective concentrations suggests that the therapeutic efficacy of atovaquone-proguanil can be considerably enhanced by the additional administration of a suitable artemisinin derivative, e.g. artesunate. PMID- 17131245 TI - Cercarial dermatitis in Austria: questionnaires as useful tools to estimate risk factors? AB - Cercarial dermatitis is a worldwide occurring skin disease characterized by itching and skin papulation. It is caused by cercariae of the fluke family Schistosomatidae. In the tropics and subtropics species of the genus Schistosoma can cause severe diseases of man. However, several genera (e.g. Trichobilharzia, Bilharziella) of medical significance are prevalent in Europe as well; they are also known as "bird schistosomes", because waterfowl is the final host. Pulmonate snails act as intermediate hosts. Humans are accidental hosts; they get infested by penetration of the cercariae into the skin of swimmers/bathers in ponds and lakes. They can not mature in humans, but die shortly after penetration. Cercarial dermatitis is known in Austria since 1969, with regularly occurrences nearly every summer. In early 2003 we created a homepage to provide information about the causative agents for the public/patients, to document the occurrences and to get data about the distribution of this parasitic disease. We therefore created a questionnaire and asked people for the following parameters: personal data, information about the waters, activity in the water and details about the dermatitis itself. A total of 34 questionnaires were returned. The results will be discussed according to their relevance as risk factors. The way how people interact with the water seems to be essential, but not demographic features. In addition, this approach revealed a new segment of the public that is at risk - owners/users of swimming ponds. PMID- 17131246 TI - [The German Ophthalmological Society (DOG) during the Period of National Socialism]. AB - Sixty-one years after the end of the Hitler dictatorship, the history of the German Ophthalmological Society (DOG) has still hardly been investigated. According to different sources, especially the reports of the DOG congresses 1934, 1936, 1938, and 1940, the following picture can be drawn: 1. The seizure of power ("Machtergreifung") of Adolf Hitler was appreciated by most of the DOG members. 2. After a change of the constitution the DOG came under the control of the "Reichsinnenministerium". However, it escaped the egalitarianism ("Gleichschaltung") and remained relatively independent. 3. Approximately 40 % of the heads of the German university eye clinics who were the most influential DOG representatives were members of the national socialistic German working party (NSDAP). Almost all of these joined the party in 1933 or later. 4. Up to the last congress in Dresden, 1940, the DOG activities were quite extensive. After that time the activities strongly declined. 5. The "Law for the prevention of genetically disabled offspring" ("Gesetz zur Verhutung erbkranken Nachwuchses") from January 1st 1934 was intensely discussed by the DOG. Some prominent ophthalmologists and DOG members were at least in part responsible for the sterilisations because of "congenital blindness". However, as far as it is known, the DOG itself did not intervene directly concerning the practice of sterilisation. 6. Between 1932 and 1940, the DOG lost approximately 12 % of its members. Many of these stemmed from foreign countries, and many were German Jews. The latter left the DOG, as Walther Lohlein stated after the end of the war, "voluntarily". However, a main reason for leaving the DOG was very likely the feeling of being unwanted. The national socialism had several disastrous effects on ophthalmology. Although single DOG members participated in the excesses, the DOG as an organization was not directly involved. However, taking into consideration that more than 10 % of the members of the pre-Hitler era left their scientific society it is a matter of interpretation whether the DOG remained completely innocent between 1933 and 1945. PMID- 17131247 TI - [How to live with keratoconus]. AB - BACKGROUND: The article is based on an undergraduate thesis that was written within the framework of investigating problems of rehabilitation in the humanities department of Berlin's Humboldt University. It deals with the issue of whether and, if so, in what spheres of life, people suffering from keratoconus may have to expect changes in lifestyle and what possibilities exist for compensation. The paper is based on the "construct of compensating impairment of sight" according to Nater, a fundamentally resource-oriented approach. Impaired vision has restrictive effects on the quality of life, which can, however, be minimised by appropriate measures of compensation so that, at best, normality can be achieved or felt. MATERIALS AND METHODS: In a Germany-wide written survey by questionnaire. The changes in lifestyle arising from the disease were registered and evaluated (n = 192). RESULTS: Ophthalmologists, optometrists and opticians attending to keratoconus patients generally emphasise the medical or optical care, respectively. The results arrived at in this study extend the medically or optically oriented view and give insights into other spheres of life of those affected, their problems and their strategies of compensation. People suffering from keratoconus occupy an intermediate position between severe dysopia and normality. The most important means of compensation for people with KC is at present the rigid gas permeable contact lens. With its help, most KC patients enjoy normal eyesight, which has a favourable effect on all the results. If keratoconus develops vehemently, restrictions in the quality of life are more severe than in the case of a slow development of the disease. Contrary to the hypothesis proposed in the paper, the existence of a tight social network does not have much of an alleviating effect on the restrictions felt. A sympathetic communication of the diagnosis, however, will have a favourable effect on the patient in accepting the condition and integrating KC into his or her life. CONCLUSIONS: In order to improve the situation of people suffering from KC, the best possible medical/optical care, the interdisciplinarity of all professions involved, and an early, sympathetic and medically well established communication of the diagnosis are essential as well as publicising the disease. PMID- 17131248 TI - [About the frequency of excessive epithelial basement membrane thickening in keratoconus]. AB - BACKGROUND: In spite of a huge amount of morphological and biochemical findings the aetiology of keratoconus (KC) is still unclear. While the primary changes are seen and found mainly within the anterior corneal stroma, some authors believe that the corneal epithelium plays the major role in KC formation. Consequently, the epithelial basement membrane (BM) has become a matter of interest in KC research. MATERIAL AND METHODS: The study included 55 consecutive patients who had a perforating keratoplasty performed because of KC. The corneal explants were processed for light microscopy in the usual manner. Slides were stained with PAS. The whole explant was investigated although the focus of interest was put on the epithelial BM. 10 normal corneas of eyes which had to be enucleated because of a choroidal melanoma served as a control. RESULTS: In 3 patients (5.5 %) there was a significant linear thickening of the BM (thickness of the BM more than 5 % but less than 30 % of the thickness of Bowman's layer). Only in one patient (1.8 %) there was an excessive BM thickening (thickness of the BM more than 30 % of that of Bowman's layer) with focal (central) BM warts directed against the epithelium ("inverse cornea guttata"). In general, thickening of the BM was most pronounced paracentrally. None of the control corneas had a significantly thickened BM. CONCLUSIONS: A massively thickened epithelial BM can be occasionally observed in KC but is rare on the whole. It is most likely an unspecific secondary phenomenon and not a prerequisite for KC development. The "inverse cornea guttata" was a unique finding. PMID- 17131249 TI - [Effect of timolol on central corneal thickness and endothelial cell density]. AB - BACKGROUND: The measurement of corneal thickness plays an increasing role in glaucoma screening and diagnosis. The influence of a variety of drugs on corneal thickness is well established. Especially for antiglaucomateous drugs this effect seems to be important. However, little is known about the influence of beta receptor antagonists on corneal thickness. The aim of this study was to provide evidence of the effect of timolol on central corneal thickness and endothelial cell density. MATERIALS AND METHODS: Ten healthy volunteers (five women and five men) with a mean age of 29 years (range 25 to 56 years) were examined in a double blind, prospective and randomised pilot study. Intraocular pressure, corneal thickness and endothelial cell density was estimated before as well as fifteen minutes, 24, 48, 72 and 96 hours after application of timolol 0.5 % eye drops twice daily. The partner eye received sodium hyaluronate eye drops twice daily and served as a control. RESULTS: The application of timolol showed a decrease of intraocular pressure from initially 12 mmHg to 9 mmHg after four days (p = 0,0188) as well as an increase of corneal thickness from 537 microm to 557 microm after four days (p = 0,0659). There was no change of intraocular pressure (p = 0,9935) or corneal thickness (p = 0,9998) in the control eyes. There was also no effect of timolol (p = 0,2782) or sodium hyaluronate (p = 0,1940) on endothelial cell density. CONCLUSIONS: The study provides evidence of the influence of beta receptor antagonists on corneal thickness. This effect may be caused by receptor mediated influences on corneal ion and fluid transport. Further studies are needed to show if the increase of corneal thickness after application of topical timolol has clinical importance. PMID- 17131250 TI - [Intraocular pressure measurements with the Proview self-tonometer in comparison of Goldmann applanation tonometry in healthy and glaucomatous eyes]. AB - BACKGROUND: The aim of this study was to compare the results of intraocular pressure (IOP) measurements obtained using the pressure phosphene tonometer Proview with those from Goldmann applanation tonometer (GAT) in normal and glaucomatous eyes. PATIENTS AND METHODS: The IOP in 150 eyes of 62 healthy volunteers and 88 patients with glaucoma or ocular hypertension was measured prospectively in a sitting position. After being trained to use the Proview device, Goldmann applanation tonometry was performed first. Then the patient took a reading with the Proview self-tonometer. RESULTS: For all investigated eyes the measurements with the Proview were on average 5.5 mmHg higher than those by GAT. Only 34 % of the readings from the two devices were within a difference range of +/- 3 mmHg. On comparing the group of glaucomatous patients with slight visual field defects with the group of healthy subjects and patients with ocular hypertension without visual field defects we determined almost the same mean difference between the Proview and GAT (mean difference in the group with visual field defects = 4.7 +/- 4.1 mmHg; without defects = 4.8 +/- 2.9 mmHg). CONCLUSIONS: The tonometer Proview did not show a close agreement to GAT. Therefore, the PPT does not offer an alternative method for measuring IOP. We do not recommend the Proview for self-tonometry at home or for clinical management of patients with glaucoma. Slight visual field defects seem to have no influence on intraocular pressure measurement with the self-tonometer. PMID- 17131251 TI - [Comparisons of manual and automatic refractometry with subjective results]. AB - BACKGROUND: Refractometry is very important in everyday clinical practice. The aim of this study is to compare the precision of three objective methods of refractometry with subjective dioptometry (Phoropter). The objective methods with the smallest deviation to subjective refractometry results are evaluated. MATERIALS AND METHODS: The objective methods/instruments used were retinoscopy, Prism Refractometer PR 60 (Rodenstock) and Auto Refractometer RM-A 7000 (Topcon). The results of monocular dioptometry (sphere, cylinder and axis) of each objective method were compared to the results of the subjective method. The examination was carried out on 178 eyes, which were divided into 3 age-related groups: 6 - 12 years (103 eyes), 13 - 18 years (38 eyes) and older than 18 years (37 eyes). All measurements were made in cycloplegia. RESULTS: The smallest standard deviation of the measurement error was found for the Auto Refractometer RM-A 7000. Both the PR 60 and retinoscopy had a clearly higher standard deviation. Furthermore, the RM-A 7000 showed in three and retinoscopy in four of the nine comparisons a significant bias in the measurement error. CONCLUSIONS: The Auto Refractometer provides measurements with the smallest deviation compared to the subjective method. Here it has to be taken into account that the measurements for the sphere have an average deviation of + 0.2 dpt. In comparison to retinoscopy the examination of children with the RM-A 7000 is difficult. An advantage of the Auto Refractometer is the fast and easy handling, so that measurements can be performed by medical staff. PMID- 17131252 TI - [Arterial ocular occlusion associated with systemic venous thrombophilia--2 case reports]. AB - BACKGROUND: Arterial ocular occlusions in apparently healthy young men are unusual. Because of inconsistent study results there is a great amount of insecurity with respect to the reasonable diagnostic approach in such a scenario. We report on the process of diagnostic steps in two young men with arterial ocular occlusion such as non-arteritic optic neuropathy (NAION) and branch retinal arterial occlusion (BRAO). CASE REPORT: Here, we report on two hitherto healthy young men who suffered from ocular arterial occlusions. Because of the apparent arterial perfusion deficit at first thrombocyte aggregation-inhibiting substances were recommended. On searching for the underlying embolic source in both men a significant arterial-venous cardiac and pulmonary shunt volume was identified. Additional screening for risk factors for venous thrombosis resulted in the identification of Factor V Leiden as well as hyperhomocysteinemia. CONCLUSION: These two cases stress the importance to exclude possible arterial venous shunts and, if identified, the need to search for additional risk factors for venous thrombosis in patients with arterial embolic ocular disease. PMID- 17131253 TI - [Changes of retinal light sensitivity after YAG-laser capsulotomy]. AB - BACKGROUND: The aim of this prospective study was firstly to investigate the changes of retinal light sensitivity of the central visual field in patients with posterior capsule opacification (PCO) after YAG-laser capsulotomy and secondly to determine the correlation between changes in retinal light sensitivity and patient's age and the diameter of posterior laser capsulotomy. PATIENTS AND METHODS: Our study includes 25 eyes (25 patients) with PCO after phacoemulsification and intraocular lens implantation. In all patients YAG-laser capsulotomy was performed. In all patients, a threshold visual field analysis was carried out with the C 30 - 2 programme of the automated Humphrey Field Analyzer before and one month after the procedure. RESULTS: In all patients a significant improvement of visual acuity was observed one month after capsulotomy (p = 0,00003). One month after YAG-laser treatment, a significant improvement of retinal light sensitivity in the central visual field was also observed. The average MD (mean deviation) before the procedure was - 6.05 db (+/- 3.2, max. - 11.7, min. + 2.17) and after the procedure - 3.61 db (+/- 3.7, max. - 10.87, min. + 0.71). Before laser capsulotomy obvious areas of reduced retinal sensitivity in 19 of 25 eyes (76 %) were found with MD p values of less than 5 %. One month after the procedure the MD p values were less than 5 % only in 8 eyes (32 %). The improvement in MD was statistically significant (p = 0,0002). No correlation was established between the improvement of retinal light sensitivity and patient's age or the size of the capsulotomy. CONCLUSIONS: Our study shows that the improvement of retinal light sensitivity is significant after YAG-laser capsulotomy and not depends on patient age or capsulotomy size. PMID- 17131254 TI - [Regression of lid-parallel conjunctival folds (LIPCOF) on topical treatment with Liposic Eye Gel--a pilot study]. AB - INTRODUCTION: The aim of this observation study was to determine whether regression of lid-parallel conjunctival folds (LIPCOF) can be induced by repeated and intensive treatment with a gel-forming tear substitute. METHODS: In cooperation with the referring ophthalmologists, patients suffering from advanced dry eye condition were recruited. LIPCOF grades from 1 to 3 were included. Tear film break-up time (BUT) had to be less than 10 sec and the Schirmer-I test less than 10 mm. All patients recruited had been using watery tear substitutes. Examinations including visual acuity, slit lamp, fluorescein test, Schirmer-I test, BUT, LIPCOF were performed at baseline and after 2 weeks and 2 months. All examinations were done by the same physician at the Dietrich-Bonhoeffer Eye Hospital of Neubrandenburg. Exclusion criteria were severe eye diseases such as glaucoma, cataract, retinal detachment, history of intraocular or conjunctival surgery and disorders of the eye lids. Pretreatment with gels was not allowed. In the study, patients were treated with Liposic Eye Gel at least three times a day for a period of two months. RESULTS: LIPCOFs regressed in 10 eyes of 5 patients (1 x male, 4 x female) from a mean grade of 2.58 at baseline to 2.29 after 2 months under Liposic treatment. This improvement is statistically significant (n = 10, p = 0.04). Schirmer-I test results had improved in the same time period from a mean of 6.4 mm at baseline to a mean of 8.1 mm after 2 months (n. s., n = 10, p = 0.73). BUT improved from a mean of 12.5 sec at baseline to a mean of 17.5 sec at the 2-month follow-up (n. s., n = 10, p = 0,085). In one patient the cornea was fluorescein-positive at baseline and stayed so throughout the follow up. Liposic Gel was used by the patients between 3 to 8 times per day. Tolerance was excellent. DISCUSSION: The study results show that the morphological signs of the dry-eye condition like LIPCOFs can be reversed by intensive treatment with Liposic Gel. The improvements in BUT and Schirmer-I test results confirm the adherence of the patients to the treatment schedule. CONCLUSION: This pilot study provides the statistical data necessary for designing a major treatment study to prove not only the reversibility of LIPCOF, but also the dependence of the possible improvement on the grade of LIPCOF stage on the age of the patient and on other criteria. PMID- 17131255 TI - [Papilloedema and brain abscess associated with isolated left superior vena cava draining in to the left atrium]. AB - BACKGROUND: Papilloedema and brain abscess associated with persistent left superior vena cava draining into the left atrium without sinus coronarius and atrium septum defect is rare and until now in only few cases have been reported. PATIENT: A 12 year-old girl was admitted because she had suffered from headache and blurred vision for 2 weeks. Fundus examination revealed papilloedema with peripapillary intraretinal haemorrhages. Pupillary reaction to light was normal. An afferent pupillary defect was negative. Neurological examination was without any focal neurological signs. Cerebral magnetic resonance imaging showed the characteristic features of a brain abscess in the left frontal lobe (3.5 x 3.0 x 2.8). Cardiological examination showed the presence of an isolated left superior vena cava draining into the left atrium in the absence of coronary sinus and atrial septum defect. CONCLUSION: Congenital heart disease such as LSVC is rare. The association of LSVC with papilloedema and brain abscess is very rarely seen. However, the complications of LSVC can be life-threatening. Therefore in children with papilloedema, intraocular and intraorbital reasons as well as congenital heart disease as the underlying reason should be included in the differential diagnosis. PMID- 17131256 TI - [Measurement of skull thickness by ultrasound]. AB - BACKGROUND: The presurgical diagnostic for measurement of skull thickness for implantations of epithesis or bone anchored hearing aids is limited on computed tomography (CT) of petrosal bone up to now. Testing of ultrasound for measurement of skull thickness is the aim of our study . MATERIAL AND METHODS: 20 patients has been included, which had got a computed tomography of nasal sinus, skull or petrosal bone. The potential localisations of implantations have been measured by 3D-reconstruction of computed tomography. The results have been statistical compared by sonographic investigations (7.5 MHz Soundhead) at the same localisation. RESULTS: The mean skull thickness has been 5,2 mm in computed tomography. Comparison has described a mean skull thickness of 4,2 mm in sonography. Statistical comparison has shown that ultrasound has been measured statistical significant too small. CONCLUSIONS: The skull thickness can be described and measured by sonography. However mean skull thickness has been measured statistical significant too small. In spite of this, conclusions relating to presurgical diagnostic, for instance for the implantation of epithesis and bone anchored hearing aids, can be drawn by sonographic data. PMID- 17131257 TI - [Temporary sensory hearing deficits after ear surgery--a retrospective analysis]. AB - BACKGROUND: Temporary and persistent threshold shifts can occur after ear surgery. So far, only few studies deal with temporary sensory hearing deficits after ear surgery. PATIENTS AND METHODS: In a retrospective study, thresholds at 500 Hz, 1000 Hz, 2000 Hz and 4000 Hz were analysed in 393 patients (125 x mesotympanal chronic otitis media, 164 x cholesteatoma, 44 x tympanosclerosis, 60 x otosclerosis) before, the first 4 days and 3 weeks after ear surgery to evaluate possible temporary threshold shifts. RESULTS: Analysis of all patients in total proved statistically significant slight temporary threshold shifts of up to 7 dB at 2000 Hz (postoperative day 1 to 3) and at 4000 Hz (postoperative day 1 and 2). Use of a drill caused a statistically slight temporary threshold shift only at 2000 Hz at the first postoperative day (4.2 dB). In cholesteatoma surgery with preparations at the ossicular chain a statistically significant slightly higher threshold level was found in all postsurgical evaluations at 500 Hz. Removal of scars or granulation tissue attached to the ossicular chain was not associated with an elevation of threshold levels. CONCLUSION: Slight temporary threshold shifts can be observed at 2000 Hz and 4000 Hz after ear surgery. Use of the drill and preparation at the ossicular chain usually results in no significant sensory hearing deficit. PMID- 17131258 TI - [Craniofacial fibrous dysplasia: scan policy or surgery?]. AB - BACKGROUND: The aetiologic correlations of fibrous dysplasia (FD) are more and more decoded by molecular biology, improved imaging procedures, and the use of computer assisted surgery--thus a review of present diagnostics and therapy methods is evaluated. METHOD: The valid methods of diagnostic and therapy procedures of craniofacial FD were retrospectively analysed in a collective of 9 patients in consideration of literature. The criteria of the decision for diagnosis and surgical procedures were evaluated. RESULTS: According to the literature, diagnosis was ascertained with modern CT and MRI scans. Bone scintigraphy was only used additionally in particular questions. In case of unclear radiological findings histomorphological procedures were used complementarily to distinguish FD from other bone tumors. The aim of surgical intervention was to reduce pain, to restore the function in compression symptoms, to recover original ostia, or to restore the natural geometry of the face. CONCLUSIONS: Current imaging procedures allow differential diagnosis from other benign bone tumors but also from malignancies. The therapy of FD is conservative (wait and scan) or operative in dependence on the localisation, the extension and the clinical manifestations of the disease. In the future molecular biological methods could function as supporting instrument for diagnosis if histomorphological results are not meaningful. PMID- 17131259 TI - [Volume reduction of tonsil hyperplasia in childhood with a surgical ultrasound device]. AB - BACKGROUND: We present the results of a prospective study concerning the use of a surgical-ultrasound device (use of ultrasound mechanical vibrations in the tonsil tissue with denaturizing effect on collagen and proteins for tonsil ablation, coablation) for the therapy of non-infectious tonsillar hyperplasia resulting in upper airway obstruction in childhood. METHOD: 51 patients with a median age of 6.9 years (3.5-12 years) were treated with surgical ultrasound, under general anesthesia. The intervention did not include tissue excision, but the denaturation of hyperplastic tonsil tissue. Intra- and postoperative bleeding, pain (by visual analog scale), tonsil-to-tonsil diameter and swallowing difficulties were under consideration. Also an bacteriological examination of tonsils was performed. RESULTS: The procedure was generally well tolerated. There was no intra- or postoperative bleeding. Swallowing difficulties were described as not very disturbing. Median follow up was 2.1 years. The postoperative "wound healing" lasted 7-9 days. A reduction of tonsil's size was visible since the first post treatment week with the final results occurring after 3-6 months. Intertonsillar distance was 9 mm preoperatively and 22.14 mm after coablation. A higher incidence of bacterial tonsillitis or development of quinsies were not reported. CONCLUSION: The treatment of tonsilar hyperplasia by means of surgical ultrasound is highly effective and causes relatively mild post surgical complaints without intra- or postoperative bleeding. PMID- 17131260 TI - [Evidence of a novel gene for the LAV-syndrome]. AB - BACKGROUND: Both LAV- (large or enlarged vestibular aqueduct) and Pendred syndrome are autosomal recessive diseases. In contrast to Pendred-syndrome, LAV syndrome is characterised only by an enlarged vestibular aqueduct. Pendred syndrome is a more complex disease. Classically it is characterised by sensorineural hearing loss and enlargement of the thyroid gland. Up to now, only mutations in SLC26A4 gene are known as being responsible for both syndromes. The gene for Pendred-syndrome (SLC26A4) has been localised by linkage analysis of chromosome 7q31. This protein is expressed in the inner ear, thyroid gland, kidney, and placenta. Functional analysis of the gene product (pendrin) in Xenopus laevis oocytes revealed that pendrin acts as an iodide/chloride and chloride/formate exchanger. METHOD: Each of the exons and flanking splice regions of the SLC26A4 gene were analysed by direct sequencing. Haplotype analysis was undertaken with microsatellite markers spanning a 5 Mbp area around the localisation of the SLC26A4 gene. RESULTS: In sequence analysis of 42 patients with bilateral enlargement of the vestibular aqueduct, no mutation could be identified in 30 % of cases. In some of these cases, a linkage to the gene localisation on chromosome 7q31 could not be detected. CONCLUSION: Our results indicate evidence for a second gene involved in the development of LAV-syndrome. PMID- 17131261 TI - [Arrosion bleeding of the brachiocephalic trunc - a late complication after percutaneous dilatative tracheostomy in a patient with a cervical rib abnormality]. AB - BACKGROUND: In the last years percutaneous dilatative tracheostomy was increasingly established in many intensive care units, mostly performed by the anesthesiologists. This method becomes problematic, when it is used in patients requiring long time intubation. It must always be kept in mind that in the course of further rehabilitation the patients may be treated by persons, who are not familiar with the altered anatomy of the upper airways. This bears the risk for severy complications, like bleeding from major vessels. CASE REPORT: Letal arrosion bleeding of the brachiocephalic trunc was observed in a patient, 3 weeks after percutaneous dilatative tracheostomy (technique by Griggs). As an abnormality the patient had a cervical rib. The wrong position of the tracheostomy tube was not noticed because the patient had no dyspnoea, spontaneously breathing through the nose and mouth instead of through the tracheostomy tube. CONCLUSIONS: Particularly in patients requiring long-time intubation it is essential that the tube can be changed safely. We advocate an "epithelialized" tracheostoma with tracheocutaneous anastomosis for these patients. If the position of the tracheostomy tube is not clear, endoscopic control has to be performed. PMID- 17131263 TI - Septal deviation 1882: the origins of systematic submucous resection of the nasal septum. 1982. PMID- 17131264 TI - Surgery of the nasal septum. AB - Long-term results after septoplasty are not satisfactory. Apart from a recurrence of deviation, there are various reasons for this: false preoperative analysis, complete straightening of the septum, and a disturbed nasal cycle. Preoperative functional diagnostics with a combination of rhinoresistometry, acoustic rhinometry, and long-term rhinoflowmetry are necessary for differentiating between "physiological" and "pathological" septal deviations and recognizing other causes for obstruction. The surgical procedure of septoplasty includes approach, mobilization, resection, reposition, and finally reconstruction of all three layers. The goal of the operation should not be complete straightening of the nasal septum. The space between the septum and turbinates is of utmost importance. It should not be enlarged as much as possible; rather, it must be shaped in such a way as to allow freely congestion and decongestion reciprocal on the two sides during the nasal cycle. PMID- 17131265 TI - Twenty-five years experience with extracorporeal septoplasty. AB - Even today, the difficult septum presents a surgical problem. A severe septum deformity is caused mostly by an accident or is seen in patients with malformations such as cleft lip and palate deformity. It is characterized by a massive malformation in all levels with consecutive blocking of one or both airways. Such marked septal deformities cannot be corrected properly by standard septoplasty techniques. Therefore, in such cases we suggest an extracorporeal septoplasty, where we take out the whole septum and reconstruct a new septal plate by different techniques followed by replantation and reconstruction of the cartilaginous dorsum. From January 1, 1981 to December 31, 2005, we operated on 2301 patients and improved this method constantly, especially the safe septal fixation and rebuilding of the cartilaginous dorsum. Follow-ups showed that even in severe deformities a revision rate of only 5% respectively 7% was found. Therefore, we conclude that the extracorporeal septoplasty with its refinements and the way we are doing it today can also be recommended to less experienced rhinosurgeons. PMID- 17131266 TI - The severely deviated septum--the way I solve the problem. AB - Surgical correction of severely deviated or "saddle" noses poses specific problems that are often difficult to resolve because of the complexity of the deformities, particularly those of the septum. Four steps are necessary: (1) the deformations must be skeletonized and visualized by a perfect extramucosal dissection and maxillary-premaxillary approaches; (2) the septum must be mobilized by resection of the osteocartilaginous chondrovomerine callus, after which it is possible to carry out the septum as long as the mucosa is not torn; (3) the angulations must be corrected carrying out discontinuous cartilaginous incisions; and (4) the structures must be stiffened. Polydioxanone (PDS) struts provide a neat solution to the issue of the reconstruction of a plane and sufficiently stiff septum. PMID- 17131267 TI - Reconstructive septal surgery. AB - To attain without compromise the surgical goal, a straight and concurrently solid septum, especially in cases of marked deviations or severe post-traumatic deformities (sometimes combined with external deformities), an excellent proven and well-known technique is extracorporal or external septoplasty. Surgery consists of excision of the quadrilateral cartilage and its division into straight fragments. To avoid postoperative sequelae such as saddling, the pieces of cartilage must be reconnected meticulously to form a straight and at the same time solid plate. This difficult and time-consuming procedure can be assisted by the use of a connecting material. Since 1996 we have been using a polydioxanone foil (PDS, Ethicon Norderstedt Germany) as the supporting material for this purpose. The PDS as well as the suture material is completely resorbed within 5 months, excluding long-term complications of other artificial implants. PMID- 17131268 TI - Evolution of the septal crossbar graft technique. AB - The septal crossbar graft is a surgical technique used to correct crooked nose and solve the associated functional and aesthetic problems. Described for the first time in 2003, it combines staggered septal incisions with a spreader graft in the dorsal septum on the concave side of the deviation. The method has proved particularly useful in straightening the septum and ensuring postoperative results of lasting stability. Clinical experience over the last few years and the identification of some snags in the procedure have prompted modifications of the technique that should be regarded essentially as evolutionary stages. This article provides a detailed description of all the surgical phases of the technique in the light of these developments and discusses its strengths with respect to the specific problems of crooked nose. Attention is drawn in this connection to both the functional effect on the internal nasal valve and the aesthetic effect of reshaping the upper lateral cartilage. PMID- 17131269 TI - Nasal valves--importance and surgical procedures. AB - One of the most difficult aspects in rhinoplasty is resolving and preventing functional compromise of the nasal valve area reliably. The nasal valves are crucial for the individual breathing competence of the nose. Structural and functional elements contribute to this complex system: the nasolabial angle, the configuration and stability of the alae, the function of the internal nasal valve, the anterior septum symmetrically separating the bilateral airways and giving structural and functional support to the alar cartilage complex and to their junction with the upper lateral cartilages, the scroll area. Subsequently, the open angle between septum and sidewalls is important for sufficient airflow as well as the position and function of the head of the turbinates. The clinical examination of these elements is described. Surgical techniques are more or less well known and demonstrated with patient examples and drawings: anterior septoplasty, reconstruction of tip and dorsum support by septal extension grafts and septal replacement, tip suspension and lateral crural sliding technique, spreader grafts and suture techniques, splay grafts, alar batten grafts, lateral crural extension grafts, and lateral alar suspension. The numerous literature is reviewed. PMID- 17131270 TI - Combining rhinoplasty with septal perforation repair. AB - A combined septal perforation repair and rhinoplasty was performed in 80 patients presenting with septal perforations (size 1 to 5 cm) and external nasal deformities. The external rhinoplasty approach was used for all cases and the perforation was repaired using bilateral intranasal mucosal advancement flaps with a connective tissue interposition graft in between. Complete closure of the perforation was achieved in 90% of perforations of size up to 3.5 cm and in only 70% of perforations that were larger than 3.5 cm. Cosmetically, 95% were very satisfied with their aesthetic result. The external rhinoplasty approach proved to be very helpful in the process of septal perforation repair especially in large and posteriorly located perforations and in cases where the caudal septal cartilage was previously resected. Our results show that septal perforation repair can be safely combined with rhinoplasty and that some of the routine rhinoplasty maneuvers, such as medial osteotomies and dorsal lowering, could even facilitate the process of septal perforation repair. PMID- 17131271 TI - Complications in septoplasty. AB - The most frequent complications of septoplasty are deformities, infections, and perforations. The effects of each of these complications, however, can be very different. Dislocations and deformities of the septum may result not only in an impaired airway but also in visible deformities of the entire nasal base and dorsum. A patient who underwent septoplasty can be "stigmatized." Infections may lead not only to septal abscess but also to endocranial complications such as meningitis or septicemia with endocarditis. Permanent perforations of the nasal septum can result in significant symptoms if they are located in the anterior part of the nose. Surgical closure is the treatment of choice, with a high success rate if the patients are selected properly. Besides these three major types of complications there are many others, from smell disturbances to blindness. Causes, prevention, and correction of selected complications are presented and data of the recent literature reported. PMID- 17131273 TI - A comparison of pressure ulcer prevalence rates in nursing homes in the Netherlands and Germany, adjusted for population characteristics. AB - Annual pressure ulcer surveys in the Netherlands and Germany have shown remarkable differences in prevalence rates. We explored the differences between the two populations, and the degree to which these differences were associated with differences in prevalence. To this end, data from 48 Dutch and 45 German facilities (n = 9772) from 2003 were analyzed. The prevalence of pressure ulcers (excluding grade 1) was 12.5% in the Netherlands and 4.3% in Germany. After adjusting for age, sex, and other risk factors, the probability of developing a pressure ulcer of stage 2 or higher in Dutch nursing homes was three times greater than in German homes. PMID- 17131274 TI - Addressing health disparities. PMID- 17131275 TI - Characteristics associated with discharge to home following prolonged mechanical ventilation: a signal detection analysis. AB - The objective of study was to identify characteristics associated with being home at 6 months in 80 patients on prolonged mechanical ventilation (PMV) (> or =7 days). At 6 months, 47.5% were home, 13.8% institutionalized, and 38.8% deceased and classified "not home." Using signal detection methodology (SDM), four mutually exclusive groups at high and low probability of being home were identified. The best outcome (94.4% home) was achieved by patients with an admission Charlson Comorbidity Score < or =3 and an Acute Physiology Score (APS) < or =21 and the worst outcome (23.4% home) by patients with an admission Charlson Comorbidity Score >3 and Health Assessment Questionnaire score >2.7. SDM provided an effective means of identifying subgroups likely to be discharged home using available information. PMID- 17131276 TI - Medication adherence patterns in adult renal transplant recipients. AB - Patient adherence to immunosuppressive medications adherence is crucial to survival of the patient and a transplanted kidney, yet adherence is variable. Using a prospective, descriptive design, immunosuppressive medication adherence of 44 renal transplant recipients was followed for 6 months at a Midwestern transplant center using electronic monitoring. Four medication adherence patterns emerged from a hierarchical cluster analysis: those who took medications on time, those who took medications on time with late/missed doses, those who rarely took medications on time and who were late with morning and/or evening doses, and those who missed doses. This study is a step toward developing and implementing interventions targeted to specific patterns of poor adherence. PMID- 17131277 TI - Thinking and/or doing as strategies for resisting smoking. AB - The effects of behavioral strategies and cognitive strategies, individually or in combination, on the likelihood of lapsing during smoking cessation were examined by random effects regression analyses of 1,499 temptations reported by 61 smokers during the first 2 weeks of cessation. Compared to using no strategies, using either type exclusively or in combination was significantly protective from lapsing. The combination was not significantly better than using multiple cognitive strategies, but was superior to using a single behavioral strategy, a single cognitive strategy, or multiple behavioral strategies. Use of coping strategies during temptation episodes was highly related to resisting smoking. Maximum benefit accrued to using more than one strategy of which at least one was a cognitive strategy. PMID- 17131278 TI - Symptoms, self-efficacy, and physical activity among individuals with multiple sclerosis. AB - Using social-cognitive theory, we examined the associations among symptoms, self efficacy, and physical activity in 196 individuals with multiple sclerosis (MS). Participants completed symptoms, self-efficacy, and physical activity measures and wore an accelerometer for 7 days. There was a moderate inverse correlation between symptoms and physical activity (varphi = -.42). Subsequent analyses indicated that symptoms had direct negative relationships with self-efficacy (gamma = -.32) and physical activity (gamma = -.24), and self-efficacy had a direct positive relationship with physical activity (beta = .57). This study provides initial support for symptoms as a correlate of physical activity in individuals with MS. Monitoring symptoms during a physical activity program and altering the exercise prescription during symptom exacerbation might be a method for increasing exercise adherence. PMID- 17131279 TI - Environmental factors as predictors of smoking among ninth-grade adolescents in Pitkaranta (Russian Karelia) and in eastern Finland. AB - The purpose of this study was to assess whether similar environmental factors predict adolescents' smoking in two different cultures: in the Pitkaranta district in Russian Karelia and in eastern Finland. The data were gathered by self-administered questionnaires from ninth-grade students in 10 comprehensive schools in Pitkaranta (n = 385) and from age-matched students in 24 schools in eastern Finland (n = 2,098). Structural equation modeling (SEM) was used to test whether similar path structures fit for boys and for girls in Pitkaranta and in eastern Finland, and to test whether regression coefficients were similar between the cultures by sex. Smoking by family members and best friends was positively related to adolescents' smoking both directly and indirectly. Environmental factors were similar predictors of smoking between the cultures for boys. For girls, different regression coefficients in Pitkaranta and in eastern Finland were found. Best friend's smoking was the most important predictor of adolescents' own smoking in every sub-sample. When indirect relationships were identified, the significance of parents' and siblings' smoking, in addition to smoking by best friends, was strongly supported. PMID- 17131280 TI - Health-related research on older inmates: an integrative review. AB - The literature on older inmates' health is fragmented and insufficiently developed. In this integrative review, 21 research articles on health and older inmates were identified, critiqued, and synthesized to determine: the minimum age criterion most commonly used; health-related variables explored; health status; the health impact of incarceration; and aging-specific policies, programs, and facilities. Age 50 and older was used most often. The top three health variables were psychiatric conditions, physical illnesses, and substance abuse. Self reports of health status varied across studies; however, inmates consistently reported health declines since incarceration. Older inmates' health needs appear often to be left unmet. Nursing investigations are needed leading to practice innovations to enhance prisoners' self-management to reduce disease burden and fiscal and societal costs. PMID- 17131281 TI - A bone health intervention for older adults living in residential settings. AB - Osteoporosis, a major health problem that increases with age, may lead to fractures and loss of independence. This study focused on testing a bone health intervention aimed at assisting older adults living independently in residential settings to maintain healthy bones with the long-term goals of fracture prevention and maintenance of independent function. Forty-seven participants (M (age) = 79.36 years) completed six 1-hour educational sessions. Pre-intervention testing of osteoporosis knowledge and healthy bone behaviors was followed by posttests at 1 week, 6 weeks, and 7(1/2) months. Findings revealed significant increases in knowledge and behaviors from baseline to 9(1/2) months, although a slight decline during the last 6 months when there was no reinforcement of learning suggests the need for periodic follow-up. PMID- 17131282 TI - The Discomfort Behavior Scale: a measure of discomfort in the cognitively impaired based on the Minimum Data Set 2.0. AB - Because persons with cognitive impairment often cannot self-report pain, it is imperative to develop instruments that use observable indicators. The purpose of this study was to develop and test the psychometric properties of the Discomfort Behavior Scale (DBS), which is comprised of 17 items from the Minimum Data Set (MDS). MDS data from 29,120 cognitively impaired nursing home residents were used for psychometric testing. Factor analyses of the DBS indicated that it reflects a single discomfort dimension. The items are tau equivalent, allowing unity weighting and simple summation to create scale scores, and the internal consistency was good. The DBS has potential to be useful in studies of efforts to improve pain management in cognitively impaired residents of nursing homes. PMID- 17131283 TI - Early predictors of post-hospital adjustment problems in critically ill young children. AB - Toddlers and preschool children have been identified as being at risk for post hospital behavioral sequelae, especially when confronted with an unanticipated intensive care unit (ICU) hospitalization. The purpose of this predictive secondary analysis was to determine demographic and clinical variables that could be assessed early during hospitalization to predict internalizing and externalizing behaviors and negative behavioral change of 2- to 7-year-old children at 3 and 6 months following an unanticipated critical care hospitalization (n = 163 mother-child dyads). The children's behavioral predilections prior to hospitalization (i.e., internalizing or externalizing behaviors) and sex, as well as elevated maternal state anxiety, marital status, and level of emotional care giving by mothers, significantly predicted child internalizing and externalizing behavior and post-hospitalization behavior changes. Results suggest several risk factors that predispose children to poor outcomes following an unanticipated hospitalization that could be targeted for intervention to improve children's emotional and behavioral health. PMID- 17131284 TI - Risk and protection in the development of problem behaviors in adolescents. AB - The development of problem behaviors among adolescents is affected by complex interactions between risk and protective factors. This study was designed to determine whether selected risk and protective factors described among participants in the National Longitudinal Study of Adolescent Health predicted problem behavior cluster membership. Approximately, 13,000 adolescents from the Add Health study were examined. Three clusters of adolescents (exhibiting normal, problem, and deviant behaviors) and changes in cluster membership over 1 year were examined for relationships to specific risk and protective factors. Findings revealed that factors for current behavior problems differ from those for changes in cluster membership. These results suggest that approaches to preventing problems may differ from those required to help adolescents who are already manifesting problems. PMID- 17131285 TI - Synthesis, conformation, and bioactivity of novel analogues of the antiviral lipopeptide halovir A. AB - We synthesized by solution-phase methods three analogues, [L-Leu(6)-OMe], [L (alphaMe)Leu(3), L-Leu(6)-OMe], and [L-(alphaMe)Val(4), L-Leu(6)-OMe] of halovir A. The [L-Leu(6)-OMe] analogue is known to be biologically equipotent to its naturally occurring, antiviral, lipopentapeptide amide parent compound. The preferred conformations of the L-(alphaMe)Leu- and L-(alphaMe)Val-containing analogues, with a potentially reinforced helicity, were compared with those of [L Leu(6)-OMe] halovir A and the natural peptide itself by use of a combination of FT-IR absorption and NMR techniques. Measurements of the antiviral activities against herpes simplex virus type-1 (HSV-1) of halovir A and its three analogues were also carried out. Interestingly, the [L-(alphaMe)Val(4), L-Leu(6)-OMe] analogue exhibits the most significant activity in reducing HSV-1 infectivity, notably higher than that of halovir A itself. PMID- 17131286 TI - Limited tendency of alpha-helical residues to form disulfide bridges: a structural explanation. AB - Disulfide bridges have an enormous impact on the structure of a large number of proteins and polypeptides. Understanding the structural basis that regulates their formation may be important for the design of novel peptide-based molecules with a specific fold and stability. Here we report a statistical analysis of the relationships between secondary structure and disulfide bond formation, carried out using a large database of protein structures. Our analyses confirm the observation sporadically reported in previous investigations that cysteine residues located in alpha-helices display a limited tendency to form disulfide bridges. The very low occurrence of the disulfide bond in all alpha-chains compared to all beta-chains indicates that this property is also evident when proteins with different topologies are investigated. Taking advantage of the large database that endorsed the analysis on relatively rare motifs, we demonstrate that cysteine residues embedded in 3(10) helices present a good tendency to form disulfide bonds. This result is somewhat surprising since 3(10) helices are commonly assimilated into alpha-helices. A plausible structural explanation for the observed data has been derived combining analyses of disulfide bond sequence separation and of the length of the different secondary structure elements. PMID- 17131287 TI - Interactions of amphipathic carrier peptides with membrane components in relation with their ability to deliver therapeutics. AB - To identify rules for the design of efficient CPPs that can deliver therapeutic agents such as nucleic acids (DNAs, siRNAs) or proteins and PNAs into subcellular compartments, we compared the properties of several primary and secondary amphipathic CPPs. Studies performed with lipid monolayers at the air-water interface have enabled identification of the nature of the lipid-peptide interactions and characterization of the influence of phospholipids on the ability of these peptides to penetrate into lipidic media. Penetration and compression experiments reveal that both peptides interact strongly with phospholipids, and observations on Langmuir-Blodgett transfers indicate that they can modify the lipid organization. Conformational investigations indicate that the lipid-peptide interactions govern the conformational state(s) of the peptides. On the basis of the ability of both peptides to promote ion permeation through both natural and artificial membranes, models illustrating the translocation processes have been proposed. One is based on the formation of a beta-barrel pore-like structure while another is based on the association of helices. PMID- 17131288 TI - Exploring interaction of beta-amyloid segment (25-35) with membrane models through paramagnetic probes. AB - The accumulation of beta-amyloid peptides into senile plaques is one of the hallmarks of Alzheimer's disease (AD). There is mounting evidence that the lipid matrix of neuronal cell membranes plays an important role in the beta-sheet oligomerization process of beta-amyloid. Abeta(25-35), the sequence of which is GSNKGAIIGLM, is a highly toxic segment of amyloid beta (Abeta)-peptides, which forms fibrillary aggregates. In the present work, two spin-labelled Abeta(25-35) analogues containing the nitroxide group of the amino acid TOAC (2,2,6,6 tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid) as a paramagnetic probe at the N- or the C-terminus of the peptide sequence, respectively, were synthesized in order to investigate the peptide-membrane interaction. The orientation and associated changes of the peptide conformation in the presence of different artificial membrane models (micelles, liposomes) were evaluated by electron paramagnetic resonance and circular dichroism techniques. The results of this study allowed us to propose a model in which the C-terminal portion of the peptide is highly associated to the membrane, while the N-terminal part extends into the aqueous phase with occasional contacts with the lipid head-group region. Interestingly, the interaction of the C-terminal portion of the peptide is particularly enhanced in the presence of sodium dodecyl sulfate (SDS) molecules. PMID- 17131289 TI - New bradykinin analogues substituted in positions 7 and 8 with sterically restricted 1-aminocyclopentane-1-carboxylic acid. AB - A sterically constrained non-coded amino acid, 1-aminocyclopentane-1-carboxylic acid (Apc), was introduced in position 7 or 8 of the bradykinin (BK) B(2) receptor antagonist, [D-Arg(0), Hyp(3), Thi(5, 8), D-Phe(7)]BK, previously synthesized by Stewart's group. This modification is believed to reduce the flexibility of the peptides, thereby forcing the peptide backbone and side chains to adopt specific orientations. Apc substitution was combined with acylation of the N-terminus with 1-adamantaneacetic acid (Aaa). The activity of four new analogues was assayed in isolated rat uterus and in rat blood pressure tests. The results clearly demonstrated that the Apc residue inserted in position 7 led to a reduction of antagonistic properties in the rat uterus assay or even restored the agonism in the blood pressure test, whereas Apc at position 8 enhanced antagonistic potency in both the tests. In both cases, acylation of the N terminus led to the enhancement of the antagonistic potency. On the basis of these findings, new potent and selective B(2) blockers might be designed. PMID- 17131290 TI - Molecular dynamics study of amyloid formation of two Abl-SH3 domain peptides. AB - Molecular dynamics (MD) simulations were carried out for two-strand and ten strand beta-sheets constructed from two peptides corresponding to the diverging turn of two homologous Abl-SH3 domains, DLSFMKGE (MK; from Drosophila) and DLSFKKGE (KK; from man), in explicit water at the temperatures of 30, 170/190 and 300 K. It was found that the 2 x MK beta-sheet is more stable than the 2 x KK beta-sheet, and that the 10 x MK beta-sheet is more stable than the 10 x KK beta sheet; this suggests that the MK systems are fibril-creating and the KK systems are not. These results might explain why most SH3 domains possess two conserved basic residues at the diverging turn, which may act as gatekeepers in order to avoid aggregation. PMID- 17131291 TI - Comparison of the stability of glutathione and related synthetic tetrapeptides by HPLC and capillary electrophoresis. AB - Glutathione and related peptides are interesting targets as protectors of biological systems against an oxidative injury. Two novel glutathione analogues, UPF1 and UPF15, have been designed and synthesised. As a result of different reactions taking place, the thiol-containing compounds oxidise to disulfides. In this study, the stability of UPF1, UPF15 and glutathione in various solutions was investigated by using HPLC and CE. The results showed that UPF1 and UPF15 are powerful hydroxyl radical scavengers and their dimerisation process velocity is higher than that of glutathione. PMID- 17131292 TI - Modulation of lung cancer growth arrest and apoptosis by Phellinus Linteus. AB - The Phellinus Linteus (PL) mushroom has been shown to possess anti-tumor properties. Through influencing lymphocytes, PL indirectly augments the host's immune system against cancer cells. PL has also been demonstrated to reduce tumor proliferation. However, the mechanisms of PL against malignant growth have not yet been fully explored. In this study, we report that PL mediates the following two activities in mouse and human lung cancer cells: cell-cycle arrest at a low concentration of PL and apoptosis in response to a high dose of PL. After exposure to a low dose of PL, G(1) growth arrest occurred in the lung cancer cells. The negative growth control mediated by PL is evidenced by the decrease of the activities of cyclin-dependent kinases CDK2, 4, and 6. In contrast, at high doses, PL-induced lung cancer cells to undergo apoptosis in a dose-dependent fashion. This was evidenced by DNA fragmentation, caspase activation, and loss of clonogenecity in the lung cancer cells, all of which were lacking in the lung cancer cells treated with low concentrations of PL as well as the normal mouse lung epithelial cells exposed to either low or high concentrations of PL. The addition of the caspase inhibitor Z-VADfmk completely suppressed PL-induced apoptosis. Furthermore, the low dose of PL was able to synergize with doxorubicin to induce apoptosis in the lung cancer cells. Thus, our findings suggest that PL regulates two responses in the lung cancer cells: cell-cycle arrest and apoptosis. PMID- 17131293 TI - Method for the synthesis of highly pure vaccines using the lipid core peptide system. AB - Traditional vaccines consisting of whole attenuated microorganisms, killed microorganisms, or microbial components, administered with an adjuvant (e.g. alum), have been proved to be extremely successful. However, to develop new vaccines, or to improve upon current vaccines, new vaccine development techniques are required. Peptide vaccines offer the capacity to administer only the minimal microbial components necessary to elicit appropriate immune responses, minimizing the risk of vaccination associated adverse effects, and focusing the immune response toward important antigens. Peptide vaccines, however, are generally poorly immunogenic, necessitating administration with powerful, and potentially toxic adjuvants. The attachment of lipids to peptide antigens has been demonstrated as a potentially safe method for adjuvanting peptide epitopes. The lipid core peptide (LCP) system, which incorporates a lipidic adjuvant, carrier, and peptide epitopes into a single molecular entity, has been demonstrated to boost immunogenicity of attached peptide epitopes without the need for additional adjuvants. The synthesis of LCP systems normally yields a product that cannot be purified to homogeneity. The current study describes the development of methods for the synthesis of highly pure LCP analogs using native chemical ligation. Because of the highly lipophilic nature of the LCP lipid adjuvant, difficulties (e.g. poor solubility) were experienced with the ligation reactions. The addition of organic solvents to the ligation buffer solubilized lipidic species, but did not result in successful ligation reactions. In comparison, the addition of approximately 1% (w/v) sodium dodecyl sulfate (SDS) proved successful, enabling the synthesis of two highly pure, tri-epitopic Streptococcus pyogenes LCP analogs. Subcutaneous immunization of B10.BR (H-2(k)) mice with one of these vaccines, without the addition of any adjuvant, elicited high levels of systemic IgG antibodies against each of the incorporated peptides. PMID- 17131294 TI - Effects of N- and C-terminal addition of oligolysines or native loop residues on the biophysical properties of transmembrane domain peptides from a G-protein coupled receptor. AB - Transmembrane domains (TMDs) of G-protein coupled receptors (GPCRs) have very low water solubility and often aggregate during purification and biophysical investigations. To circumvent this problem many laboratories add oligolysines to the N- and C-termini of peptides that correspond to a TMD. To systematically evaluate the effect of the oligolysines on the biophysical properties of a TMD we synthesized 21 peptides corresponding to either the second (TPIFIINQVSLFLIILHSALYFKY) or sixth (SFHILLIMSSQSLLVPSIIFILAYSLK) TMD of Ste2p, a GPCR from Saccharomyces cerevisiae. Added to the termini of these peptides were either Lys(n) (n = 1,2,3) or the corresponding native loop residues. The biophysical properties of the peptides were investigated by circular dichroism (CD) spectroscopy in trifluoroethanol-water mixtures, sodium dodecyl sulfate (SDS) micelles and dimyristoylphosphocholine (DMPC)-dimyristoylphosphoglycerol (DMPG) vesicles, and by attenuated total reflection Fourier transform infrared (ATR-FTIR) in DMPC/DMPG multilayers. The results show that the conformation assumed depends on the number of lysine residues and the sequence of the TMD. Identical peptides with native or an equal number of lysine residues exhibited different biophysical properties and structural tendencies. PMID- 17131295 TI - 'Click peptide': a novel 'O-acyl isopeptide method' for peptide synthesis and chemical biology-oriented synthesis of amyloid beta peptide analogues. AB - After over a decade of studies on aspartic protease inhibitors and water-soluble prodrugs, we have been developing a novel method, since 2003, called 'O-acyl isopeptide method', for the synthesis of peptides containing difficult sequences. With our recent discoveries of 'O-acyl isodipeptide unit' and the 'racemization free segment condensation method', this method has further evolved as a general synthetic method for peptides. Moreover, 'Click Peptide', which could be a powerful tool for identifying the pathological functions of amyloid beta peptides in Alzheimer's disease, represents a valuable use of the isopeptide method in Chemical Biology-oriented research. PMID- 17131296 TI - Proceedings from the 10th Naples Workshop on Bioactive Peptides, June 11-14, 2006. PMID- 17131297 TI - Synthesis, characterization and hybridization studies of new nucleo-gamma peptides based on diaminobutyric acid. AB - In the present work, we report the synthesis and the characterization of a new chiral nucleoaminoacid, in which a diaminobutyric moiety is connected to the DNA nucleobase by an amidic bond, and its oligomerization to give the corresponding nucleo-gamma-peptide. The ability of this synthetic polymer to bind complementary DNA was studied in order to explore its possible use in antigene/antisense or diagnostic applications. Our interest in the presented DNA analogue was also supported by the importance of gamma-aminoacid-containing compounds in natural products of biological activity and by the known stability of gamma-peptides to enzymatic degradation. Furthermore, our work could contribute to the study of the role of nucleopeptides as prebiotic material in a PNA world that could successively lead to the actual DNA/RNA/protein world, as recently assumed. PMID- 17131298 TI - Effect of salts on the structural behavior of hPrP alpha2-helix-derived analogues: the counterion perspective. AB - Both theoretical studies and direct experimental evidence have emphasized the importance of electrostatic interactions in the general phenomenon of spontaneous amyloid fibril formation. A number of observations have recently spurred interest in the contribution of these interactions to the conformational behavior of the prion protein. In this paper, we show how salt addition and pH change can modify the conformation of two peptide analogues derived from the human prion protein helix 2 according to a Hofmeister-series-type dependence. Employment of various sodium salts allowed us to highlight the fact that chaotropic anions favor unstructured conformation, whereas kosmotropic anions promote the formation of compact structures like alpha-helix and beta-sheet, which may ultimately facilitate fibril formation. This finding should warn people engaged in ion-based research on prion and derived peptides about cation-bound effects, which have been almost exclusively investigated to date, being easily confounded with modifications that are actually caused by anion activity, thus leading researchers into misunderstand ion-specific effects. To avoid the common complication of ion confounding, it is highly desirable that experiments be designed so that the species causing the modification can be unequivocally perceived. PMID- 17131299 TI - Probing DNA-peptide interaction forces at the single-molecule level. AB - The versatility of chemical peptide synthesis combined with the high sensitivity of AFM single-molecule force spectroscopy allows us to investigate, quantify, and control molecular recognition processes (molecular nanotechnology), offering a tremendous potential in chemical biology.Single-molecule force spectroscopy experiments are able to detect fast intermediate transition states, details of the energy landscape, and structural changes, while avoiding multiple binding events that can occur under ensemble conditions. Dynamic force spectroscopy (DFS) is even able to provide data on the complex lifetime. This minireview outlines the biophysical methodology, discusses different experimental set-ups, and presents representative results in the form of two case studies, both dealing with DNA-binding peptides. They may serve as model systems, e.g., for transcription factors or gene transfection agents. PMID- 17131300 TI - Aldehyde modification of peptide immunogen enhances protein-reactive antibody response to toxic shock syndrome toxin-1. AB - Introduction of aldehyde groups into protein conjugates enhanced the immune response to a coupled peptide without the use of strong adjuvants. Synthetic peptides representing the N-terminal (residues 1-16) and internal (residues 53 65) epitopes of toxic shock syndrome toxin-1 (TSST-1) were coupled to carrier protein, and carbonyl tags were introduced by Amadori reaction with glycolaldehyde. Modified and unmodified antigens in alum were used to immunize rabbits and the reactivities of antisera were compared. Aldehyde modification augmented the response detected by ELISA, which included enhanced binding to peptides and to native TSST-1. In western blot, TSST-1 was detected by antiserum elicited to the N-terminal peptide, but not that generated to the peptide representing the internal sequence. The same antiserum also neutralized TSST-1 activity in a lymphocyte proliferation assay. The circular dichroism spectrum of the N-terminal peptide indicated a propensity for helical conformation, similar to the structure at the corresponding sequence of the native protein. These data suggest that aldehyde modification can boost immunogenicity of peptide-based vaccines, generating epitope-specific immune responses against the cognate protein antigens without using potent adjuvants. PMID- 17131301 TI - Factors influencing Hong Kong Chinese patients' decision-making in seeking early treatment for acute myocardial infarction. AB - The purpose of this study was to identify, through in-depth interview, factors that influenced 27 Hong Kong Chinese patients' decision-making in seeking early treatment for acute myocardial infarction (AMI). The median delay time from the onset of symptoms to arrival at the hospital was 15.6 hours for men and 53.7 hours for women. Three major categories emerged from the data: (a) becoming aware of the threat, (b) maintaining a sense of normality, and (c) struggling to mobilize resources. A variety of decisions were made by patients from the onset of chest pain to seeking help. These decisions were heavily influenced by healthcare factors (access to emergency medical service (EMS) and treatment), personal factors (cognitive interpretations of symptoms), sociocultural factors (family situation, cultural beliefs, and practices), and coping strategies. PMID- 17131302 TI - Does adding psychotherapy to pharmacotherapy improve social functioning in the treatment of outpatient depression? AB - The existence of an overall association between severity of depression and level of social functioning is well documented. To increase the probability of a long term recovery, a normal level of social functioning is essential. It is currently unknown whether combined therapy has a better outcome than pharmacotherapy with regard to social functioning. In a 6-month randomized clinical trial in outpatients with major depression, all patients studied had a baseline score of at least 14 points on the 17-item Hamilton Depression Rating Scale (HDRS). The two conditions consist of pharmacotherapy (PhT) (N=84) and combined therapy (CoT), pharmacotherapy plus 16 sessions of short psychodynamic supportive psychotherapy (N=83). Efficacy was assessed using the 17-item HDRS, the Clinical Global Impression (CGI) Severity and Improvement scales, the Depression subscale of the Symptom Checklist-90 (SCL-90), the Quality of Life Depression Scale (QLDS), more the Groningen Social Disability Schedule (GSDS). Severity of depression decreased significantly (on the SCL-90 Depression subscale and the QLDS) more in the CoT condition. A larger improvement in social functioning was demonstrated for remitted patients than for nonremitted patients. The number of dimensions of social functioning that had improved significantly was higher in CoT than in PhT. There was a moderate advantage of the CoT condition on both depressive symptoms and level of social functioning in comparison with PhT. We also found a positive association between depression severity and level of social functioning. PMID- 17131303 TI - Are there gender differences in catastrophic appraisals in panic disorder with agoraphobia? AB - Our aim in this study was to compare panic-related catastrophic appraisals between women and men with panic disorder with agoraphobia (PDA). One hundred two outpatients with PDA (75 women and 27 men) participated. Two instruments for the assessment of catastrophic appraisals, Agoraphobic Cognitions Questionnaire and Panic Appraisal Inventory, were administered before and after cognitive behavioral therapy (CBT) that also included pharmacotherapy in three-fourths of the patients. Female and male patients did not differ significantly in terms of their tendency to anticipate catastrophic consequences of panic, before or after CBT-based treatment. For both females and males, the tendency to make catastrophic appraisals decreased significantly with treatment. We conclude that among patients with PDA there are no gender differences in catastrophic appraisals of panic sensations and symptoms. The apparently higher risk of panic recurrence in women does not seem to be related to their panic-related catastrophic appraisals. These findings also support a notion that there is no gender difference in response to CBT-based treatment of PDA. PMID- 17131304 TI - Prominent expression of metalloproteinases in early stages of ovarian tumorigenesis. AB - The role for matrix metalloproteinases (MMPs) in tumor cells invasion and metastasis is well established, and expression of MMPs is recognized as an indication of tumor cell malignancy. Previous studies suggest that the degradation of the basement membrane is a crucial early step in epithelial transformation and ovarian tumorigenesis. Thus, MMPs may also express and exert a role in preneoplastic lesions of ovarian tissues. We investigated the expression of the major metalloproteinases, gelatinase A, 72 kDa type IV collagenase (MMP 2), and gelatinase B, 92 kDa type IV collagenase (MMP-9), and the presence of basement membrane in ovarian tumors and tissues from prophylactic oophorectomies using immunostaining. MMP expression was also characterized in a panel of ovarian cancer cell lines and several nontumorigenic ovarian surface epithelial primary cells by zymography, Northern, and Western blots. We found, surprisingly, that MMP-2 and MMP-9 are expressed more frequently in early lesions than in established carcinomas. No correlation was found between the expression of MMPs and tumor grades or stages. In preneoplastic lesions, MMP-2 or MMP-9 expression often associates with the absence of basement membrane and morphological alterations. MMP-2 is often expressed in nontumorigenic ovarian surface epithelial cells but reduced or absent in cancer cells. Thus, we conclude that MMPs expression does not correlate with the malignancy of ovarian epithelial cells as generally thought. Rather, increased metalloproteinase expression is an early event in ovarian tumorigenesis and associates with the loss of epithelial basement membrane and morphological transformation. We propose that the increased MMP activity is an etiological factor for ovarian cancer risk. We found that MMPs expression does not correlate with the malignancy of ovarian epithelial cells as generally thought. Rather, increased metalloproteinase expression is an early event in ovarian tumorigenesis. The finding suggests roles of MMP in tumor initiation in addition to invasion, and may impact on the strategy for use of MMP inhibitors in cancer prevention. PMID- 17131305 TI - 17beta-Estradiol differentially regulates androgen-responsive genes through estrogen receptor-beta- and extracellular-signal regulated kinase-dependent pathways in LNCaP human prostate cancer cells. AB - The molecular mechanism underlying the actions of estrogens in normal prostate physiology and prostate cancer development remains unclear. In the present study we tested the hypothesis that estrogens modulate androgen-dependent events in prostate cells by examining the effects of 17beta-estradiol (E2) on androgen responsive genes (ARGs) in the androgenresponsive LNCaP cells. We found that LNCaP cells express estrogen receptor-beta (ER-beta) as the major form of ER and ER treatment with E2 led to an increase in cell growth. The proliferative effect of E2 correlated with induction of several ARGs by E2. Interestingly, some other ARGs did not respond to E2. Consistent with involvement of ER-beta, the induction of both cell growth and ARG mRNA levels by E2 was attenuated by the pure antiestrogen ICI 182,780. Moreover, we found ER-beta small interfering RNA attenuated induction of ARG mRNAs by E2. However, the effect of E2 on ARG mRNA appeared also to require the androgen receptor and to be mediated through activation of the extracellular-signal regulated kinase (ERK) pathway. These results provide mechanistic evidence supporting a direct effect of estrogen, mediated through ER-beta- and ERK-dependent pathways, on specific molecular targets in human prostate cancer cells. PMID- 17131306 TI - Prevention of hyperthermia with silk of the oriental hornet, Vespa orientalis: a hypothesis. AB - Wasps apparently develop normally even under extreme thermal conditions, including deserts. We deemed it worthwhile to set up an experiment wherein wasp brood combs containing a full gamut of brood ranging from eggs up to pupae and a few adults were kept in an incubator whose temperature was gradually raised to 45 degrees C, and the response of the disparate brood to such warming was photographed via Infra Red camera. The finding of this experiment showed that for open brood (i.e., eggs, larvae at various instars, and empty cells) the temperature was close to the ambient temperature, but in the silk coated pupae, the temperature was lower than the ambient by up to 4 degrees C. This lower temperature was retained for at least 90 min of incubation. For comparison we evaluated the relative contribution of the pupae to the phenomenon, by warming also a vacant, (i.e., a broodless and silkless comb) in parallel to a comb from which the pupae had been extricated but the silk weave retained and left behind. We found that the totally empty comb heated up under these conditions to nearly 110 degrees C, whereas the silk-containing vacant cells only heated up to about 40 degrees C. These finding are discussed from two aspects, namely the importance for wasps to maintain a constant temperature throughout the pupating process, and the manner in which the silk weave contributes to such a goal. PMID- 17131307 TI - Risk-based selection from the general population in a screening trial: selection criteria, recruitment and power for the Dutch-Belgian randomised lung cancer multi-slice CT screening trial (NELSON). AB - A method to obtain the optimal selection criteria, taking into account available resources and capacity and the impact on power, is presented for the Dutch Belgian randomised lung cancer screening trial (NELSON). NELSON investigates whether 16-detector multi-slice computed tomography screening will decrease lung cancer mortality compared to no screening. A questionnaire was sent to 335,441 (mainly) men, aged 50-75. Smoking exposure (years smoked, cigarettes/day, years quit) was determined, and expected lung cancer mortality was estimated for different selection scenarios for the 106,931 respondents, using lung cancer mortality data by level of smoking exposure (US Cancer Prevention Study I and II). Selection criteria were chosen so that the required response among eligible subjects to reach sufficient sample size was minimised and the required sample size was within our capacity. Inviting current and former smokers (quit 15 cigarettes/day during >25 years or >10 cigarettes/day during >30 years was most optimal. With a power of 80%, 17,300-27,900 participants are needed to show a 20-25% lung cancer mortality reduction 10 years after randomisation. Until October 18, 2005 11,103 (first recruitment round) and 4,325 (second recruitment round) (total = 15,428) participants have been randomised. Selecting participants for lung cancer screening trials based on risk estimates is feasible and helpful to minimize sample size and costs. When pooling with Danish trial data (n = +/-4,000) NELSON is the only trial without screening in controls that is expected to have 80% power to show a lung cancer mortality reduction of at least 25% 10 years after randomisation. PMID- 17131308 TI - Doxycycline induces caspase-dependent apoptosis in human pancreatic cancer cells. AB - Doxycycline (DC) belongs to the tetracycline family of antibiotics and has been used clinically for over 5 decades. Despite advances in understanding the molecular pathogenesis of pancreatic cancer, no chemotherapy course has shown significant effectiveness. Hence new treatments are needed. In this study we report the pro-apoptotic effects of DC in 2 pancreatic adenocarcinoma cell lines, T3M4 and GER. Cell proliferation was measured using the SRB protein dye. Induction of apoptosis was detected using ELISA. Caspase activation was detected using either immunoblotting or a colorimetric assay based on cleavage of caspase associated substrates. Expression of proteins and post-translational modifications were determined using immunoblotting. Treatment of pancreatic cancer cells with DC reduces their proliferation. This reduction is, at least partly, due to increased caspase-dependent apoptosis involving activation of caspase3, caspase7, caspase8, caspase9, caspase10 and increased levels of FADD. Inhibition of caspase8 or caspase10 but not caspase9 significantly decreases DC induced apoptosis in both cell lines. Furthermore treatment of pancreatic cancer cells with DC increases protein levels of Bax and phosphorylation of members of the p38MAPK pathway such as p38MAPK, MKK3/6 and MAPKAPK2. These results provide an insight into mechanisms behind the pro-apoptotic effects of DC in pancreatic cancer cells. PMID- 17131309 TI - Dietary polyphenolic phytochemicals--promising cancer chemopreventive agents in humans? A review of their clinical properties. AB - Epidemiological and preclinical evidence suggests that polyphenolic phytochemicals exemplified by epigallocatechin gallate from tea, curcumin from curry and soya isoflavones possess cancer chemopreventive properties. Whilst such naturally occurring polyphenols have been the subject of numerous mechanistic studies in cells, information on their clinical properties, which might help assess their promise as human cancer chemopreventive agents, is scarce. Therefore, we present a review of pilot studies and trials with a cancer chemoprevention-related rationale, in which either healthy individuals or patients with premalignant conditions or cancer received polyphenolic phytochemicals. The review identifies trial design elements specifically applicable to polyphenolic phytochemicals. The available evidence for tea polyphenols tentatively supports their advancement into phase III clinical intervention trials aimed at the prevention of progression of prostate intraepithelial neoplasia, leukoplakia or premalignant cervical disease. In the case of curcumin and soya isoflavones more studies in premalignacies seem appropriate to optimise the nature and design of suitable phase III trials. The abundance of flavonoids and related polyphenols in the plant kingdom makes it possible that several hitherto uncharacterised agents with chemopreventive efficacy are still to be identified, which may constitute attractive alternatives to currently used chemopreventive drugs. PMID- 17131310 TI - In vitro and in vivo molecular evidence of genistein action in augmenting the efficacy of cisplatin in pancreatic cancer. AB - We recently reported the potential of genistein in augmenting gemcitabine-induced killing of pancreatic cancer (Banerjee, S. et al., Cancer Research 2005;65:9064 72). Since cis-diaminedichloroplatinum (II) (cisplatin) is widely used against solid tumors, we further investigated whether genistein pretreatment could be used as a novel strategy for cisplatin-induced killing of pancreatic cancer cells in vitro and enhanced antitumor activity in vivo. Our in vitro results showed that pretreatment of cells with genistein followed by cisplatin resulted in significant loss of cell viability and potentiated apoptosis irrespective of the metastatic ability of cells. Mechanistically, genistein augmented cisplatin induced killing by down regulating transcription factor-NF-kappaB and anti apoptotic Akt expression. NF-kappaB was found upregulated when pancreatic cancer cells were exposed to cisplatin, suggesting the potential mechanism of acquired chemo-resistance. In addition, we also showed, for the first time, that genistein in combination with cisplatin is more effective antitumor agent in our orthotopic tumor model. But most importantly, our data also showed that a specific target, such as NF-kappaB, was inactivated in animal tumors treated with genistein and cisplatin. Immunohistochemical data showed reduced staining for phospho-p65, Bcl xL and MMP-9 in treated tumors compared to control tumors, but the lowest activity was seen in the combination group. These results provide strong molecular in vivo evidence in support of our hypothesis that inactivation of the NF-kappaB signaling pathway by genistein results in the chemo-sensitization of pancreatic tumors to cisplatin, which is likely to be an important and novel strategy for the treatment of pancreatic cancer. PMID- 17131311 TI - Monitoring a national cancer prevention program: successful changes in cervical cancer screening in the Netherlands. AB - The success of screening, an important cancer prevention tool, depends on the quality and efficiency of protocols and guidelines for screening and follow-up. However, even centrally organized screening programs such as the Dutch cervical screening program occasionally show problems in performance. To improve this program, the screening scheme, follow-up, administration and financing protocols and guidelines were thoroughly changed in 1996. This study evaluates the consequences for the performance of the national program. Five-year coverage rate, the proportion of screened women sent to follow-up, follow-up compliance and duration, and the yearly number of Pap smears before and after the changes in 1996 were compared. Five-year coverage increased substantially in the added target age groups (30-34, and 54-60 years); in the old target age group (35-53 years) it remained around 80%. The percentage of screened women sent to follow-up decreased from almost 19-3% per screening round, due to a more restrictive use of the Pap 2 classification, and an evidence-based cessation of follow-up of negative smears without endocervical cells. Follow-up compliance has improved, and the average time until a woman is either referred or rejoins the regular screening schedule, has become shorter. The total number of smears, a strong determinant of screening costs, has decreased by 20% primarily due to the changed follow-up recommendations. In conclusion, the 1996 changes in protocols and guidelines, and their implementation have increased coverage and efficiency, and decreased the screening-induced negative side effects. PMID- 17131312 TI - Human papillomavirus seropositivity and risks of head and neck cancer. AB - We examined antibody response to VLP HPV-16, HPV-16 E6 and E7 antibodies as potential seromarkers of HPV-related head and neck cancer (HNC). The study included 204 HNC cases and 326 controls evaluated for HPV presence in sera using ELISAs for anti-HPV VLP antibodies and HPV-16 E6 and/or E7 antibodies, and in tumor tissue using PCR and DNA sequencing. Anti-HPV-16 VLP was detected in 33.8% of cases and 22.4% of controls, anti-E6 in 20.6% of cases and 0.9% of controls and anti-E7 in 18.6% of cases and 0.6% of controls. HPV-16 DNA was detected in 26.1% of tumors. The adjusted risk of HNC was elevated among those seropositive for HPV-16 VLP (odds ratio (OR) = 1.7, 1.1-2.5), E6 (OR = 32.8, 9.7-110.8) or E7 (OR = 37.5, 8.7-161.2). Compared to HPV DNA-negative/seronegative cases, tumor HPV-16 cases had increased risk of detection with anti-VLP antibodies (OR = 6.8, 3.1-14.9). The odds were more pronounced among cases seropositive for E6 (OR = 69.0, 19.3-247) or E7 (OR = 50.1, 14.7-171). Antibodies against E6 or E7 were associated with risk of cancer in the oral cavity (OR = 5.1, 1.2-22.4) and oropharynx (OR = 72.8, 16.0-330), and with disease characteristics: stage, grade and nodal status. Anti-E6 and/or E7 antibodies were found in 74% of tumor HPV-16 positive cases but in only 5% of tumor HPV-negative cases (K =0.7, 0.6-0.8) suggesting good correlation between the serologic marker and HPV tumor status. Antibodies to HPV-16 E6 and/or E7 represent a more specific biomarker than anti HPV-16 VLP of an HPV-related HNC. Because of the survival advantage of HPV related HNC, HPV-16 E6/E7 detection may be useful in therapy targeted for HPV related tumors. PMID- 17131313 TI - Statins and risk of cancer: a systematic review and metaanalysis. AB - We conducted a systematic review of the association between HMG-CoA reductase inhibitor (statin) use and cancer risk. We searched MEDLINE, EMBASE, Web of Science, ISI Proceedings and BIOSIS Previews bibliographic databases, electronic trials registers and reference lists for potentially eligible randomized trials and observational studies. Thirty-eight individual studies (26 randomized trials involving 103,573 participants and 12 observational studies with 826,854 participants) were included. Median follow-up was 3.6 and 6.2 years for trials and observational studies, respectively. In metaanalyses of randomized trials, there was no evidence that statin therapy was associated with incidence of all cancers (26 trials; pooled risk ratio = 1.00; 95% CI 0.95-1.05; I(2) = 0%) or the following site-specific cancers: breast (7 trials; risk ratio = 1.01; 0.79-1.30; I(2) = 43%), prostate (4 trials; risk ratio = 1.00; 0.85-1.17; I(2) = 0%), colorectum (9 trials; risk ratio = 1.02; 0.89-1.16; I(2) = 0%), lung (9 trials; risk ratio = 0.96; 0.84-1.09; I(2) = 0%), genito-urinary (5 trials; risk ratio = 0.95; 0.83-1.09; I(2) = 0%), melanoma (4 trials; risk ratio = 0.86; 0.62-1.20; I(2) = 17%) or gastric (1 trial; risk ratio = 1.00; 0.35-2.85). There was no evidence of differential effects by length of follow-up, statin type (lipophilic vs. lipophobic) or potency. Trial results were generally consistent with observational studies. We conclude that statin use is not associated with short term cancer risk, but longer-latency effects remain possible. PMID- 17131314 TI - Dimeric ansamycins--a new class of antitumor Hsp90 modulators with prolonged inhibitory activity. AB - The geldanamycin derivative 17-allyamino-17-demethoxygeldanamycin (17-AAG) is a clinical stage ATP-competitive HSP90 inhibitor that induces degradation of HSP90 client proteins. 17-AAG contains 1 ansamycin moiety and is highly potent in conventional cell killing assays. Since active Hsp90 exists as a dimer, we hypothesized that dimeric compounds containing 2 ansamycin pharmacophores might inhibit Hsp90 function more efficiently than 17-AAG. Here, we show that monomeric and dimeric ansamycins exert their activity in distinct ways. Under conditions of continuous exposure, 17-AAG induced client degradation and cell growth inhibition more readily than the dimeric drugs CF237 and CF483. By contrast, 24 hr treatment of various tumor cells with 17-AAG followed by drug washout caused temporary client degradation and cell cycle arrest but minimal cell death, whereas both dimers induced massive apoptosis. CF237 remained bound to Hsp90 for days after drug withdrawal and, while both monomeric and dimeric compounds caused accumulation of the inactive intermediate Hsp90 complex, this effect disappeared following washout of 17-AAG but not CF237. The dimer was also retained for longer in tumor xenografts and displayed superior antitumor activity in vivo. These results indicate that monomeric and dimeric Hsp90 inhibitors have distinct biological profiles and work differentially toward target inhibition. PMID- 17131315 TI - Body size, hormone therapy and risk of breast cancer in Asian-American women. AB - Historically, breast cancer rates have been low in Asia but rates have increased substantially in Asian-Americans for reasons that are not well understood. The authors conducted a population-based case-control study of breast cancer in Los Angeles County, which included 1,277 (450 Chinese, 352 Japanese, 475 Filipinos) women with incident, histologically confirmed breast cancer and 1,160 control subjects (486 Chinese, 311 Japanese, 363 Filipinos). A detailed in-person interview was conducted, which included questions on menopausal hormone therapy (HT) use, height, weight in each decade of life and reproductive factors. Breast cancer risk increased with increasing recent weight in postmenopausal women (p trend = 0.015). There was a significant 16% (95% CI = 2-35%) increase in risk per 10 kg of body weight in postmenopausal women. In both premenopausal and postmenopausal women, risk increased with increasing waist to hip ratio; this remained statistically significant after adjustment for recent weight in all subjects combined (p trend = 0.042). The increased risk associated with high recent weight in postmenopausal women was more apparent for women with high waist to hip ratio (p trend = 0.013). Use of HT was a significant risk factor; risk increased 26% per 5 years of current use of estrogen and progestin therapy (p trend = 0.017). The increased risk associated with high body weight was observed irrespective of HT use. Use of HT and high body size might have contributed to the rapid increase of breast cancer in Asian-Americans. PMID- 17131316 TI - Time trends of esophageal cancer in Hong Kong: age, period and birth cohort analyses. AB - This study was to examine the time trend of the incidence rates of esophageal cancer during the period 1979-2003 in Hong Kong and to identify the effects of year of diagnosis (period) and year of birth (cohort) on the observed time trends using regression models. Cancer incidence data were obtained from Hong Kong Cancer Registry and population data were from the Census and Statistics Department. Age-standardized incidence rates were computed by the direct method using the World population of 1966. Annual percentage change (APC) in incidence rate was calculated using nonlinear regression. Period and cohort effects were assessed using 2 separate Poisson regression models after adjusting for age. During the period 1979-2003, a steady decrease in the age-standardized incidence rate was observed for both males (APC = -3.38%, 95% confidence interval [CI]: 2.89%, -3.86%) and females (APC = -3.92%, 95% CI: -3.15%, -4.69%). The incidence rates were consistently higher among males than females. After the adjustment for age and with the period 1989-1993 or birth cohort of 1934-1938 as reference, the relative risk of more recent periods or birth cohorts significantly decreased. The age-cohort model provided a better description of the data than the age period model. Given reasonable latency between exposures and esophageal cancer incidence, the declining birth cohort effects in the recent generations were in line with the increased intakes of fresh vegetables and decreased consumptions of alcohol drinking, tobacco smoking, and preserved foods observed in population, thus supported their importance in influencing the burden of esophageal cancer. PMID- 17131317 TI - CagA+ Helicobacter pylori infection and gastric cancer risk in the EPIC-EURGAST study. AB - Helicobacter pylori (H. pylori), atrophic gastritis, dietary and life-style factors have been associated with gastric cancer (GC). These factors have been evaluated in a large case-control study nested in the European Prospective Investigation into Cancer and Nutrition carried out in 9 countries, including the Mediterranean area. Participants, enrolled in 1992-1998, provided life-style and dietary information and a blood sample (360,000; mean follow-up: 6.1 years). For 233 GC cases diagnosed after enrolment and their 910 controls individually matched by center, gender, age and blood donation date H. pylori antibodies (antilysate and antiCagA) and plasma Pepsinogen A (PGA) were measured by ELISA methods. Severe chronic atrophic gastritis (SCAG) was defined as PGA circulating levels <22 microg/l. Overall, in a conditional logistic regression analysis adjusted for education, smoke, weight and consumption of total vegetables, fruit, red and preserved meat, H. pylori seropositivity was associated with GC risk. Subjects showing only antibodies anti-H. pylori lysate, however, were not at increased risk, while those with antiCagA antibodies had a 3.4-fold increased risk. Overall, the odds ratio associated with SCAG was 3.3 (95% CI 2.2-5.2). According to site, the risk of noncardia GC associated with CagA seropositivity showed a further increase (OR 6.5; 95% CI 3.3-12.6); on the other hand, a ten fold increased risk of cardia GC was associated with SCAG (OR 11.0; 95% CI 3.0 40.9). These results support the causal relationship between H. pylori CagA+ strains infection, and GC in these European populations even after taking into account dietary habits. This association was limited to distal GC, while serologically defined SCAG was strongly associated with cardia GC, thus suggesting a divergent risk pattern for these 2 sites. PMID- 17131318 TI - Breast cancer guidelines--Physicians' intentions and behaviors. AB - Guidelines are written to define what a physician should do, and networks set up to provide every patient with good practice. However, is willingness to treat according norms enough to actually implement it? Between 1997 and 2003, 4,533 women with invasive, noninflammatory, nonmetastatic breast cancer have been treated within the framework of a regional network (R2C). The rate of implementation of 5 consensual norms was assessed. The rate of "abnormal" management regarding surgical re-excision for inadequate margin was found to be 12.6%. The main explanatory variable was patient age >70 years (OR = 4.05). For nodal exploration, the sampling quality threshold was set at 10. Mean rate of lack of compliance was 25.2%. The 2 main explicative factors were surgeon's experience and women's age. The observed rate of "insufficient" irradiation dose was 18.2%. The main explanatory variables were age (with a gradient) and a negative nodal status. Concerning adjuvant chemotherapy, the rate of no treatment (despite consensual indication) was 16.0%. Again, the main explicative factor was age (with a gradient). Women's age appears to be a major explanatory variable predicting lack of physician's compliance with consensual norms. Besides the age of the women, a "better" prognosis (negative nodal status and pT < or = 20 mn) is often associated with lack of compliance. It is not clear, however, if it's the rules that do not fit the clinical situation of aging patients or the physicians who are not aware of the benefit of consensual disease management for aging patients. PMID- 17131319 TI - The effects of aging on tumor growth and angiogenesis are tumor-cell dependent. AB - It is generally accepted that histologically similar tumors grow more slowly, with less angiogenesis, in aged mice relative to young mice. We subcutaneously implanted TRAMP-C2 tumor cells, a prostate cancer cell line not previously examined in aging, into syngeneic C57/Bl6 young (4 month) and aged (20 month) mice and compared tumor growth and angiogenesis. Unexpectedly, the prostate tumors grew as fast in aged as in young mice. Angiogenesis in TRAMP-C2 tumors was robust, with no differences between the young and aged mice in the number of vessels, distribution of vessel sizes or features of vessel maturation. Aged mice had lower levels of serum testosterone than the young mice. VEGF levels were similar in the tumors and sera of the young and aged mice. Comparison with B16/F10 melanoma, a cancer cell line that is representative of previous studies in aged mice, showed that B16/F10 tumors grew minimally in the aged mice. In contrast to the B16/F10, TRAMP-C2 tumors had an extracellular matrix with significantly higher levels of MMP2 and MMP9 expression and activity. These unique results demonstrate that tumor progression can be as robust in aged tissues as young tissues. The ability of aged mice to grow large, vascularized prostate tumors is associated with high levels of MMP2/9 activity that may produce a permissive environment for tumor growth and angiogenesis. These data demonstrate that tumor-cell specific features determine the effect of aging on tumor growth and angiogenesis. PMID- 17131320 TI - Socioeconomic inequality and short-term outcome in Hodgkin's lymphoma. AB - Socioeconomic status (SES) is a determinant of outcome in various types of cancer. The aim of this study is to analyze the impact of the SES in Hodgkin's lymphoma (HL). From 2001 to 2005, 194 consecutive patients were prospectively followed in 5 institutions. Patients answered a questionnaire with a set of items used to determine the SES, and were then divided in 2 groups according to their SES score. There were 151 patients (78%) with a higher SES and 43 patients (22%) with a lower SES. The complete remission (CR) rate was 82%. Patients with a higher SES had a higher CR rate than those with a lower SES (85 vs. 72%, crude odds ratio = 2.27, p = 0.046). A lower SES and the performance status >1 were independently associated with a trend towards a lower CR, even when controlled for the other covariables of interest. Ten patients (5%) died during treatment. Death during treatment was associated with a lower SES (16 vs. 2%, p = 0.001), a performance status >1 (p < 0.0001), a lower lymphocyte count (p = 0.012) and weakly with a lower albumin level (p = 0.065). With a median follow-up of 1.7 years, a higher SES was associated with a better 2-year overall survival (93 vs. 79%, p = 0.01). In underprivileged countries, patients with a lower SES require a more careful monitoring during treatment, possibly with specific support measures. Regimens more intense than doxorubicin, bleomycin, vinblastine and dacarbazine could pose a prohibitive risk of complications in this group of patients. (c) 2006 Wiley-Liss, Inc. PMID- 17131321 TI - Up-regulation of Mac-2 binding protein by hTERT in gastric cancer. AB - Mac-2 binding protein (Mac-2BP) is a secreted tumor antigen that is elevated in many cancers and implicated in tumor metastasis, as well as cell adhesion and immune functions. We focused on the human telomerase reverse transcriptase (hTERT) induced Mac-2BP expression and the relationship between Mac-2BP expression and the progression of gastric cancer. A cDNA expression array analysis was performed on the telomerase-negative cell line, SW13, which was engineered to overexpress hTERT when compared with the parental SW13 cell. hTERT induced Mac-2BP expression was confirmed via RT-PCR and Northern blotting. ELISA and flow cytometric analyses revealed that Mac-2BP protein was increased by 2- to 4-fold in hTERT-overexpressing cells compared with the mock control. Mac-2BP expression was significantly reduced when the overexpressed hTERT was neutralized by the introduction of hTERT-specific siRNA. These results suggest that Mac-2BP expression is modulated by hTERT. Mac-2BP levels in both gastric cancer cells and tumor tissues were determined via Northern blot analysis and immunohistochemistry. Mac-2BP protein was highly expressed in most gastric cancer cell lines, and gastric tumor tissues were stained more densely than normal tissues. The intracellular and secreted Mac-2BP levels were also evaluated via ELISA, indicating that Mac-2BP was expressed and secreted more abundantly in gastric cancer patients than in healthy donors. The elevated serum Mac-2BP level in gastric tumor patients was also significantly associated with distant metastasis (p = 0.05) and higher tumor stage (p = 0.04). Our findings suggest that Mac-2BP is induced by hTERT, and that it may prove to be a useful prognostic marker for the detection of malignant progression of metastatic stomach cancers. PMID- 17131322 TI - Emmprin in epithelioid sarcoma: expression in tumor cell membrane and stimulation of MMP-2 production in tumor-associated fibroblasts. AB - Emmprin is a transmembrane glycoprotein on tumor cells that stimulates peritumoral fibroblasts to produce matrix metalloproteinases (MMPs). Emmprin and the induced MMPs play a crucial role in tumor progression, invasion and metastasis of human carcinomas (epithelial malignancies). However, only a few reports have addressed its role in soft tissue sarcomas. This study investigated the expression and role of emmprin in epithelioid sarcoma (ES). Immunoblot studies of 2 ES cell lines showed that they express emmprin, and co-culture of these ES cells with dermal fibroblasts resulted in upregulation of gelatinase A (MMP-2) in fibroblasts, as shown by zymography, immunoblotting and enzyme immunoassay. This stimulation was inhibited by an activity-blocking peptide against emmprin and by antiemmprin antibody. In addition, in vivo, immunohistochemical analysis of 5 ES patient cases demonstrated diffuse emmprin expression in ES cells and MMP-2 expression in both ES cells and peritumoral fibroblasts. The histopathological findings that peritumoral fibroblasts that were not in direct contact with emmprin-expressing ES cells exhibit upregulated MMP-2 prompted us to look for a soluble form of emmprin. Soluble full-length emmprin released from ES cells was detected in conditioned medium and shown to stimulate MMP-2 production by fibroblasts. In conclusion, emmprin is expressed in ES in both membrane and soluble forms and stimulates MMP-2 production via interactions with fibroblasts, which could play a role in ES cell stromal invasion and vascular involvement. PMID- 17131323 TI - Comparison of risk factors for invasive squamous cell carcinoma and adenocarcinoma of the cervix: collaborative reanalysis of individual data on 8,097 women with squamous cell carcinoma and 1,374 women with adenocarcinoma from 12 epidemiological studies. AB - Squamous cell carcinomas account for about 80% of cancers of the uterine cervix, and the majority of the remainder are adenocarcinomas. There is limited evidence on the extent to which these histological types share a common etiology. The International Collaboration of Epidemiological Studies of Cervical Cancer has brought together and combined individual data on 8,097 women with invasive squamous cell carcinoma, 1,374 women with invasive adenocarcinoma and 26,445 women without cervical cancer (controls) from 12 epidemiological studies. Compared to controls, the relative risk of each histological type of invasive cervical cancer was increased with increasing number of sexual partners, younger age at first intercourse, increasing parity, younger age at first full-term pregnancy and increasing duration of oral contraceptive use. Current smoking was associated with a significantly increased risk of squamous cell carcinoma (RR = 1.50, 95% CI: 1.35-1.66) but not of adenocarcinoma (RR = 0.86 (0.70-1.05)), and the difference between the two histological types was statistically significant (case-case comparison p < 0.001). A history of screening (assessed as having had at least one previous nondiagnostic cervical smear) was associated with a reduced risk of both histological types, but the reduction was significantly greater for squamous cell carcinoma than for adenocarcinoma (RR = 0.46 (0.42-0.50) and 0.68 (0.56-0.82), respectively; case-case comparison, p = 0.002). A positive test for cervical high-risk HPV-DNA was a strong risk factor for each histological type, with 74% of squamous cell carcinomas and 78% of adenocarcinomas testing positive for HPV types 16 or 18. Squamous cell and adenocarcinoma of the cervix share most risk factors, with the exception of smoking. PMID- 17131324 TI - Different types of postmenopausal hormone therapy and mammographic density in Norwegian women. AB - Postmenopausal hormone therapy (HT) is associated with increased risk of breast cancer. The HTs used in Scandinavia is associated with higher risk estimates than those used in most other countries. Mammographic density is one of the strongest risk factors for breast cancer, and possibly an intermediate marker for breast cancer. We decided to examine the relationship between use of different types of HT and mammographic density in Norwegian women. Altogether, 1,007 postmenopausal participants in the governmental mammographic screening program were asked about current and previous HT use. Mammograms were classified according to percent and absolute mammographic density. Overall, current users of HT had on average 3.6% higher mean percent mammographic density when compared with never users (p < 0.001). After adjustment for age at screening, number of children and BMI in a multivariate model, women using the continuous estradiol (E(2)) plus norethisterone acetate (NETA) combination had a mean percent mammographic density significantly higher than never users (6.1% absolute difference). Those using the continuous E(2) plus NETA combination had an 4.8% (absolute difference) higher mean percent mammographic density after <5 years of use when compared with never users, while the corresponding number for >or=5 years of use was 7% (p-trend < 0.001). We found similar associations when absolute mammographic density was used as the outcome variable. In summary, our study shows a statistical significant positive dose-response association between current use of the continuous E(2) plus NETA combination and both measures of mammographic density. PMID- 17131325 TI - Quantitative 19F MR spectroscopy at 3 T to detect heterogeneous capecitabine metabolism in human liver. AB - Chemotherapy in non-responding cancer patients leads to unnecessary toxicity. A marker is therefore required that can predict the sensitivity of a specific tumour to chemotherapy, which would enable individualisation of therapy. 19F MR spectroscopy (19F MRS) can be used to monitor the metabolism of fluorinated drugs. The aim of this study was to develop a method for quantified localised detection of fluorinated compounds in human liver. For this purpose, sensitivity optimised localised 19F MRS methods at 3 T were used to detect MR signals from capecitabine, 5'DFUR, 5'DFCR and FBAL after oral intake of capecitabine. As the radio-frequency (rf) coil is made tuneable to 19F and 1H, the same localisation method is applied to obtain 1H MR signals of water and of the 19F metabolites. In addition, T1 measurements have been performed to correct for measurement-induced saturation effects. Finally, absolute tissue concentrations of capecitabine metabolites were obtained in vivo, which revealed a substantial spatial heterogeneity of these metabolites in human liver after chemotherapy. PMID- 17131326 TI - Relation between goiter and autoimmune thyroid disease, and gastric cancer. PMID- 17131327 TI - Reproductive risk factors for incident bladder cancer: Iowa Women's Health Study. AB - We studied the association between reproductive factors and bladder cancer incidence in a prospective cohort study of 37,459 Iowa women aged 55-69 years and initially free from cancer in 1986. Women reported reproductive history and were followed prospectively through 2003. After adjusting for age and smoking, there was an inverse association between age at menopause and incident bladder cancer (n = 192). Compared with menopause at age > or =48, the hazard ratio (HR) of bladder cancer was 1.32 (95% CI; 0.90-1.94) for menopause at 43-47, and 1.60 (95% CI; 1.06-2.39) for < or =42 (p-trend = 0.02). The associations were similar for ages at natural and surgical menopause. In addition, women with a history of bilateral oophorectomy had an increased risk of bladder cancer compared with those who did not undergo bilateral oophorectomy: HR = 1.58 (95% CI; 1.12, 2.22). Finally, there was an indication of a positive association between bladder cancer and shorter lifetime years of ovulation (p-trend = 0.09). There were no associations between incident bladder cancer and age at first birth, number of births, age at menarche, use of hormone replacement therapy or any other reproductive characteristics. This study provides evidence that increased risk of bladder cancer is associated with earlier age at menopause in postmenopausal women. PMID- 17131328 TI - Involvement of ribonucleotide reductase M1 subunit overexpression in gemcitabine resistance of human pancreatic cancer. AB - Pancreatic cancer is the most lethal of all solid tumors partially because of its chemoresistance. Although gemcitabine is widely used as a first selected agent for the treatment of this disease despite low response rate, molecular mechanisms of gemcitabine resistance in pancreatic cancer still remain obscure. The aim of this study is to elucidate the mechanisms of gemcitabine resistance. The 81-fold gemcitabine resistant variant MiaPaCa2-RG was selected from pancreatic cancer cell line MiaPaCa2. By microarray analysis between MiaPaCa2 and MiaPaCa2-RG, 43 genes (0.04%) were altered expression of more than 2-fold. The most upregulated gene in MiaPaCa2-RG was ribonucleotide reductase M1 subunit (RRM1) with 4.5-fold up-regulation. Transfection with RRM1-specific RNAi suppressed more than 90% of RRM1 mRNA and protein expression. After RRM1-specific RNAi transfection, gemcitabine chemoresistance of MiaPaCa2-RG was reduced to the same level of MiaPaCa2. The 18 recurrent pancreatic cancer patients treated by gemcitabine were divided into 2 groups by RRM1 levels. There was a significant association between gemcitabine response and RRM1 expression (p = 0.018). Patients with high RRM1 levels had poor survival after gemcitabine treatment than those with low RRM1 levels (p = 0.016). RRM1 should be a key molecule in gemcitabine resistance in human pancreatic cancer through both in vitro and clinical models. RRM1 may have the potential as predictor and modulator of gemcitabine treatment. PMID- 17131329 TI - Loss of chromosomal integrity drives rat mammary tumorigenesis. AB - Breast cancer incidence varies with diet and other environmental influences, including carcinogen exposure. However, the effects of carcinogens on cell growth control pathways are poorly understood. Here, we have examined processes that are activated in the mammary glands of rats treated with 1-methyl-1-nitrosourea (MNU). This synthetic carcinogen was used to study events occurring during mammary tumor initiation and development. In female Wistar-Furth rats, given 1 dose of MNU beginning at 50 days of age, 84% of the rats developed tumors by 46 weeks of age (latency 13-15 weeks). Changes in the gland occurred as early as 1 day post-MNU. Cells exhibited DNA damage, leading to chromosomal instability, supernumerary centrosomes and higher levels of Aurora A; these events correlated with the appearance of preneoplasia in the glands. In mammary tumors, elevated numbers of centrosomes coincided with genomic instability. Tumors were transplanted into syngeneic hosts and subsequent tumor generations displayed the same marker chromosomes in mostly aneuploid metaphases with hyperdiploid numbers of chromosomes, suggesting that clonality and aneuploidy were passed on from one generation to the next. Collectively, these data suggest that the carcinogen MNU induces changes resulting in genetic instability detectable before hyperplasia and tumors develop in the rat mammary gland. PMID- 17131330 TI - Risk of second malignant neoplasms among lymphoma patients with a family history of cancer. AB - Radiotherapy and chemotherapy are known risk factors for second cancers after lymphoma. The role of genetic influences, however, remains largely unknown. We assessed risk of second cancers associated with family history of any cancer in 41,181 patients with Hodgkin lymphoma (HL) (n = 7,476), non-Hodgkin lymphoma (NHL) (n = 25,941), or chronic lymphocytic leukemia (CLL) (n = 7,764), using a large population-based database. Family history of cancer was based on a diagnosis of any cancer in 110,862 first-degree relatives. We found increased relative risk (RR) (1.81, 95% confidence interval (CI): 1.04-3.16) of breast cancer among HL patient with positive (vs. negative) family history of cancer. Among CLL patients with positive (vs. negative) family history of cancer, we observed elevated risks of bladder (RR = 3.53, 95% CI: 1.31-9.55) and prostate cancer (RR = 2.15, 95% CI: 1.17-3.94). For NHL patients with positive (vs. negative) family history of cancer, we observed non-significantly increased risk of non-melanoma skin cancer (RR = 1.94, 95% CI: 0.86-4.38) and lung cancer (RR = 1.99, 95% CI: 0.73-5.39). Our observations suggest that genetic factors, as measured by positive family history of cancer, may be influential risk-factors for selected second tumors following lymphoproliferative disorders. PMID- 17131331 TI - Carbohydrate intake, glycemic index and glycemic load in relation to risk of endometrial cancer: A prospective study of Swedish women. AB - The associations of carbohydrate intake, glycemic index and glycemic load with endometrial cancer risk were examined among 61,226 participants of the Swedish Mammography Cohort who were cancer-free at enrollment between 1987 and 1990 and completed a food frequency questionnaire. During a mean follow-up of 15.6 years, through June 2005, 608 incident cases of endometrial adenocarcinoma were diagnosed. We observed no overall association between carbohydrate intake, glycemic index or glycemic load and incidence of endometrial cancer; the rate ratios (RRs) for the highest versus the lowest quintile were 1.12 (95% CI, 0.85 1.47) for carbohydrate intake, 1.00 (95% CI, 0.77-1.30) for glycemic index and 1.15 (95% CI, 0.88-1.51) for glycemic load. However, among obese women (body mass index, BMI > or =30 kg/m2), endometrial cancer incidence was nonsignificantly elevated in the top versus bottom quintiles of carbohydrate intake (RR, 1.68; 95% CI, 0.86-3.29) and glycemic load (RR, 1.57; 95% CI, 0.82-2.99). In a subanalysis of women who completed a follow-up questionnaire in 1997, which collected information on physical activity, carbohydrate intake and glycemic load were positively related to endometrial cancer risk among overweight women (BMI > or =25 kg/m2) with low physical activity. In this subgroup, the multivariate RRs comparing extreme quartiles were 1.90 (95% CI, 0.84-4.31) for carbohydrate intake and 2.99 (95% CI, 1.17-7.67) for glycemic load. Results from this cohort study suggest that a high carbohydrate intake and a high glycemic load may increase the risk of endometrial cancer among overweight women with low physical activity. PMID- 17131332 TI - Elucidation of a protein signature discriminating six common types of adenocarcinoma. AB - Pathologists are commonly facing the problem of attempting to identify the site of origin of a metastatic cancer when no primary tumor has been identified, yet few markers have been identified to date. Multitumor classifiers based on microarray based RNA expression have recently been described. Here we describe the first approximation of a tumor classifier based entirely on protein expression quantified by two-dimensional gel electrophoresis (2DE). The 2DE was used to analyze the proteomic expression pattern of 77 similarly appearing (using histomorphology) adenocarcinomas encompassing 6 types or sites of origin: ovary, colon, kidney, breast, lung and stomach. Discriminating sets of proteins were identified and used to train an artificial neural network (ANN). A leave-one-out cross validation (LOOCV) method was used to test the ability of the constructed network to predict the single held out sample from each iteration with a maximum predictive accuracy of 87% and an average predictive accuracy of 82% over the range of proteins chosen for its construction. These findings demonstrate the use of proteomics to construct a highly accurate ANN-based classifier for the detection of an individual tumor type, as well as distinguishing between 6 common tumor types in an unknown primary diagnosis setting. PMID- 17131333 TI - SV40 and human cancer: a review of recent data. AB - An unknown proportion of formalin-inactivated poliovirus vaccine lots administered to millions of US residents between 1955 and 1963 was contaminated with small amounts of infectious simian virus 40 (SV40), a polyomavirus of the rhesus macaque. It has been reported that mesothelioma, brain tumors, osteosarcoma and non-Hodgkin lymphoma (NHL) contain SV40 DNA sequences and that SV40 infection introduced into humans by the vaccine probably contributed to the development of these cancers. The Immunization Safety Review Committee of the Institute of Medicine (IOM) reviewed this topic in 2002. The present review of recent studies showed that the earlier results describing the recovery of SV40 DNA sequences from a large proportion of the above tumors were not reproducible and that most studies were negative. Contamination with laboratory plasmids was identified as a possible source of false positive results in some previous studies. The low-level immunoreactivity of human sera to SV40 was very likely the result of cross-reactivity with antibodies to the SV40-related human polyomaviruses BKV and JCV, rather than of authentic SV40 infection. SV40 sero reactivity in patients with the suspect tumors was no greater than that in controls. In epidemiologic studies, the increased incidence of some of the suspect tumors in the 1970s to 1980s was not related to the risk of exposure to SV40-contaminated vaccines. In summary, the most recent evidence does not support the notion that SV40 contributed to the development of human cancers. PMID- 17131334 TI - Dietary zinc, copper and selenium, and risk of lung cancer. AB - Zinc, copper and selenium are important cofactors for several enzymes that play a role in maintaining DNA integrity. However, limited epidemiologic research on these dietary trace metals and lung cancer risk is available. In an ongoing study of 1,676 incident lung cancer cases and 1,676 matched healthy controls, we studied the associations between dietary zinc, copper and selenium and lung cancer risk. Using multiple logistic regression analysis, the odds ratios (OR) and 95% confidence intervals (CI) of lung cancer for all subjects by increasing quartiles of dietary zinc intake were 1.0, 0.80 (0.65-0.99), 0.64 (0.51-0.81), 0.57 (0.42-0.75), respectively (p trend = 0.0004); similar results were found for men. For dietary copper, the ORs and 95% CI for all subjects were 1.0, 0.59 (0.49 0.73), 0.51 (0.41-0.64), 0.34 (0.26-0.45), respectively (p trend < 0.0001); similar reductions in risk and trend were observed by gender. Dietary selenium intake was not associated with risk, except for a significant inverse trend (p = 0.04) in men. Protective trends (p < 0.05) against lung cancer with increased dietary zinc intake were also found for all ages, BMI > 25, current smokers, pack years < or =30, light drinkers and participants without emphysema. Increased dietary copper intake was associated with protective trends (p < 0.05) across all ages, BMI, smoking and vitamin/mineral supplement categories, pack-years < or =30 and 30.1-51.75 and participants without emphysema. Our results suggest that dietary zinc and copper intakes are associated with reduced risk of lung cancer. Given the known limitations of case-control studies, these findings must be interpreted with caution and warrant further investigation. PMID- 17131335 TI - The association of use of sunbeds with cutaneous malignant melanoma and other skin cancers: A systematic review. AB - Exposure to solar ultraviolet (UV) radiation is a known cause of skin cancer. Sunbed use represents an increasingly frequent source of artificial UV exposure in light-skinned populations. To assess the available evidence of the association between sunbed use and cutaneous malignant melanoma (melanoma) and other skin cancers, a systematic review of the literature till March 2006 on epidemiological and biological studies on sunbed use was performed in Pubmed, ISI Web of Science, Embase, Pascal, Cochrane library, Lilacs and Medcarib. Search for keywords in the title and in the abstract was done systematically and supplemented by manual searches. Only case-control, cohort or cross-sectional studies were selected. Data were abstracted by means of a standardized data-collection protocol. Based on 19 informative studies, ever-use of sunbeds was positively associated with melanoma (summary relative risk, 1.15; 95% CI, 1.00-1.31), although there was no consistent evidence of a dose-response relationship. First exposure to sunbeds before 35 years of age significantly increased the risk of melanoma, based on 7 informative studies (summary relative risk, 1.75; 95% CI, 1.35-2.26). The summary relative risk of 3 studies of squamous cell carcinoma showed an increased risk. For basal cell carcinoma, the studies did not support an association. The evidence does not support a protective effect of the use of sunbeds against damage to the skin from subsequent sun exposure. Young adults should be discouraged from using indoor tanning equipment and restricted access to sunbeds by minors should be strongly considered. PMID- 17131336 TI - Mitofilin and titin as target antigens in melanoma-associated retinopathy. AB - Melanoma-associated retinopathy (MAR) is a rare paraneoplastic syndrome in patients with melanoma. Since the onset of MAR symptoms is often associated with tumor progression or recrudescence of metastases, MAR-related symptoms are prognostic relevant. The pathomechanism underlying MAR is supposed to result from antibody production against yet unknown melanoma-associated antigens that are also expressed in retinal tissue, leading to the destruction of retinal cells and resulting in defective signal transduction. Only a 35 kDa protein in Muller glial cells, a 22 kDa neuronal antigen and retinal transducin have been identified as MAR-associated antigens to date. To identify additional antigens potentially involved in the pathogenesis of MAR, we screened a retina cDNA phage library for reactivity with antibodies in the sera from 9 patients with MAR or subclinical MAR using the serological analysis of recombinantly expressed clones (SEREX) approach. Six sera from melanoma patients without evidence of MAR and 10 sera from healthy donors served as controls. Mitofilin and titin were identified as antigens against which antibodies were found exclusively in sera of MAR patients, but not in the sera of MM patients without MAR or healthy donors. This is the first study to demonstrate that titin is highly expressed from retinal tissue and melanoma. The fact that none of the MAR-associated antigens detected to date by their capacity to elicit a humoral immune response is located on the cell surface questions a major pathogenetic role of the respective antibodies and suggests that cellular, rather than humoral mechanisms are operative in the primary immune attack against the retina in MAR. PMID- 17131337 TI - MTHFR C677T and colorectal cancer risk: A meta-analysis of 25 populations. AB - The common functional methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism may influence the risk of colorectal cancer (CRC), but data from published studies with individually low statistical power are conflicting. To clarify the role of MTHFR C677T genotype in CRC, we considered all available studies in a meta-analysis. Studies reporting on MTHFR C677T genotype and CRC were searched in PubMed up to April 2006. The principle prior hypothesis was that homozygosity for MTHFR 677TT would be associated with reduced risk of CRC. Data were available for 29,931 subjects, including 12,243 with CRC, from 25 independent populations. Compared to the homozygous CC genotype, the MTHFR 677TT genotype was associated with a reduced risk of CRC (odds ratio (OR): 0.83; 95% confidence interval (CI): 0.75-0.93; p = 0.001). There was some heterogeneity among the results of individual studies, but this was not statistically significant (heterogeneity p = 0.12; I2 = 25.8%). Heterozygosity for MTHFR 677 did not influence CRC risk (OR: 0.99; 95% CI: 0.94-1.04). These findings indicate that individuals homozygous for the MTHFR 677TT genotype are at moderately reduced risk of CRC, and support the proposal that common genetic variation in the MTHFR gene contributes to CRC susceptibility, probably accounting for at least 9% of the total incidence. PMID- 17131338 TI - The 6-maleimidocaproyl hydrazone derivative of doxorubicin (DOXO-EMCH) is superior to free doxorubicin with respect to cardiotoxicity and mitochondrial damage. AB - Doxorubicin causes a chronic cardiomyopathy in which genetic and functional lesions of mitochondria accumulate in the long-term and explain in part the delayed onset of heart dysfunction. DOXO-EMCH a 6-maleimidocaproyl hydrazone derivative of doxorubicin, is an albumin binding prodrug which has entered clinical trials because of its superior antitumor and toxicological profile. In the present work, we examined the chronic cardiotoxicity of DOXO-EMCH in direct comparison with doxorubicin. Rats (11 weeks of age) were treated with intravenous doxorubicin (0.8 mg/kg weekly for 7 weeks), an equimolar dose of DOXO-EMCH (1.1 mg/kg), or with 3.3 mg/kg of DOXO-EMCH. Controls received saline. Animals were euthanized at 48th week. Rats exposed to doxorubicin had a severe clinical, and histopathological cardiomyopathy with depressed myocardial activity of cytochrome c-oxidase (COX, 26% of controls), reduced expression of the mtDNA-encoded COX II subunit, decreased mtDNA copy numbers (46% of controls), and high levels of malondialdehyde and superoxide (787% of controls). All parameters were highly correlated with myocardial damage. Both DOXO-EMCH groups did not differ from controls with regard to clinical symptomatology, mortality and mitochondrial enzymes, although the myocardia of the high-dose group had slightly increased histopathological abnormalities, depressed mtDNA copies (74% of controls) and elevated superoxide levels (347% of controls). Doxorubicin-exposed hearts and to a lesser extent the myocardia of both DOXO-EMCH groups contained mtDNA-deletions. In summary both DOXO-EMCH doses were superior over doxorubicin with respect to clinical and histopathological evidence of cardiomyopathy, myocardial COX activity, COX II expression, mtDNA-content, mtDNA mutation loads and superoxide production in rats. PMID- 17131339 TI - Prediction of metastasis from low-malignant breast cancer by gene expression profiling. AB - Promising results for prediction of outcome in breast cancer have been obtained by genome wide gene expression profiling. Some studies have suggested that an extensive overtreatment of breast cancer patients might be reduced by risk assessment with gene expression profiling. A patient group hardly examined in these studies is the low-risk patients for whom outcome is very difficult to predict with currently used methods. These patients do not receive adjuvant treatment according to the guidelines of the Danish Breast Cancer Cooperative Group (DBCG). In this study, 26 tumors from low-risk patients were examined with gene expression profiling. An intermediate risk group of 34 low-malignant T2 tumors that fulfilled all other low-risk criteria than tumor size was included to increase statistical power. A 32-gene classifier, HUMAC32, was identified and it predicted metastases with 80% sensitivity and 77% specificity. The classifier was also validated in an independent group of high-risk tumors resulting in comparable performance of HUMAC32 and a 70-gene classifier developed for this group. Furthermore, the 70-gene signature was tested in our low- and intermediate risk samples. The results demonstrated high cross-platform consistency of the classifiers. Higher performance of HUMAC32 was demonstrated among the low malignant cancers compared with the 70-gene classifier. This suggests that although the metastatic potential to some extend is determined by the same genes in groups of tumors with different characteristics and risk, expression-based classification specifically developed in low-risk patients have higher predictive power in this group. PMID- 17131340 TI - A novel copper complex of 3-benzoyl-alpha methyl benzene acetic acid with antitumor activity mediated via cyclooxygenase pathway. AB - Pancreatic cancer (PC) is characterized as one of the deadliest malignancies and its treatment is a great challenge to clinical oncologists. Expression of COX-2 is detectable in 75% of PCs among which 50% showed overexpression, suggesting the importance of COX-2 enzyme and its metabolic product prostaglandin E2 (PGE(2)) in PC. Here the authors report the synthesis and biological activity of a novel COX 2 inhibitor, FPA-306, and its effects on PC cells with different levels of COX-2 expression. Using MTT assay, the authors found a significant growth inhibition of BxPC-3 cells treated by FPA-306 with an IC(50) of 10 micromol/L, which was lower than that of ketoprofen (IC(50) = 35.4 micromol/L) and celecoxib (IC(50) > 100 micromol/L). There was no such effect found in MIAPaCa cell line, which does not express COX-2. The authors also found dose dependent reduction in cell survival and induction of apoptosis by FPA-306 treatment in BxPC-3 cells but not in MIAPaCa cells. These results were correlated with apoptosis data and secreted PGE(2) levels. The molecular modeling of FPA-306 in the COX-2 active site showed that FPA-306 is potentially able to inhibit the activity of enzyme by blocking the active site, thereby resulting in decreased PGE(2) production. The authors also found a significant reduction of COX-2 at the mRNA and protein levels together with downregulation of NF-kappaB DNA binding activity and its downstream genes, Bcl-2 and survivin. These results suggest that FPA-306 is an effective and potent agent in inhibiting the growth of PC cells. PMID- 17131341 TI - Combining high selectivity of replication via CXCR4 promoter with fiber chimerism for effective adenoviral oncolysis in breast cancer. AB - Conditionally replicative adenoviruses (CRAds) represent novel therapeutic agents that have been recently applied in the context of breast cancer therapy. However, deficiencies in the ability of the adenovirus to infect target tumor cells and to specifically replicate within the tumor target represent key deficiencies preventing the realization of the full potential of this therapeutic approach. Minimal expression of the adenovirus serotype 5 (Ad5) receptor CAR (coxsackie and adenovirus receptor) on breast cancer cells represents a major limitation for Ad5 based virotherapy. Genetic fiber chimerism is a method to alter the tropism of Ad5-based CRAds to achieve CAR-independent infectivity of tumor cells. Here, we describe the use of a CRAd with cancer specific transcriptional control of the essential Ad5 E1A gene using the human CXCR4 gene promoter. We further modified the fiber protein of this agent by switching the knob domain with that of the adenovirus serotype 3. The oncolytic activity of this 5/3 fiber-modified CRAd was studied in breast cancer cell lines, primary breast cancer and human liver tissue slices from patients, and in a xenograft breast cancer mouse model. This infectivity enhanced CRAd agent showed improved replication and killing in breast cancer cells in vitro and in vivo with a remarkable specificity profile that was strongly attenuated in nonbreast cancer cells, as well as in normal human breast and liver tissues. In conclusion, utilization of a CRAd that combined infectivity enhancement strategies and transcriptional targeting improved the CRAd-based antineoplastic effects for breast cancer therapy. PMID- 17131342 TI - Identification of HLA-A24 restricted shared antigen recognized by autologous cytotoxic T lymphocytes from a patient with large cell carcinoma of the lung. AB - The aim of the present study was to elucidate the tumor-specific cellular immunological responses occurring in a patient with large cell carcinoma of the lung who had no evidence of recurrence following surgical resections of both a primary lung lesion and a metastatic adrenal lesion. We analyzed an autologous tumor-specific cytotoxic T lymphocytes (CTL clone F2b), which were HLA-A*2402 restricted from regional lymph node lymphocytes. The F2b possessed T cell receptor (TCR) using the Valpha5 and Vbeta7 gene segment. The existence of precursor CTL (pCTL) against autologous tumor cells (A904L) was analyzed using CTL clone-specific PCR. Lymphocytes with the same TCR as F2b were detected in the primary tumor tissue, regional lymph node and the peripheral blood collected from the patient 3 years after the operation. Using the F2b, we identified a cDNA clone encoding the tumor antigen using cDNA expression cloning method. The gene was found to encode splicing variant of the Tara gene. Finally, we identified the 9-mer Ag peptide, using constructions of mini-genes. The F2b recognized 3 out of 7 HLA-A24 positive allogeneic tumor cell lines and in 1 out of 7 HLA-A24 negative allogeneic tumor cell lines when transfected with HLA-A24. This peptide is therefore considered to be potentially useful for performing specific immunotherapy in a significant proportion of lung cancer patients bearing HLA A24. PMID- 17131343 TI - Pyrazolo-pyrimidine-derived c-Src inhibitor reduces angiogenesis and survival of squamous carcinoma cells by suppressing vascular endothelial growth factor production and signaling. AB - Src tyrosine kinase family cooperates with activated growth factor receptors to regulate growth, invasion and metastasis. The authors examined the influence of a novel c-Src inhibitor, 1l, derived from 4-amino-substituted-pyrazolo-pyrimidines, on tumor angiogenesis and on the angiogenic output of squamous carcinoma cells, A431 and SCC-4. The effect of 1l was assessed on growth and microvessel density in A431 tumors and its effect compared with the established c-Src inhibitor PP-1. The effects of c-Src inhibition were investigated on vascular endothelial growth factor (VEGF) expression and activity in tumor cells grown in vivo and in vitro, as well as on VEGF mediated signaling and on endothelial cell functions. Nanomolar concentrations of 1l decreased tumor volume promoted by A431 implanted in nude mice, without affecting in vitro cell tumor survival. This effect was related to 1l inhibition of VEGF production, and secondary to an effect on tumor microvessel density. The rabbit cornea assay confirmed that 1l markedly decreased neovessel growth induced by VEGF. In cultured endothelial cells, 1l inhibited the VEGF-induced phosphorylation on tyr416 of c-Src, resulting in a reduced cell proliferation and invasion. Consistently, 1l dowregulated endothelial nitric oxide synthase, MAPK-extracellular receptor kinase 1-2 (ERK1-2) activity and matrix metalloproteinases (MMP-2/MMP-9), while the tissue inhibitors of metalloproteinases (TIMP2/TIMP-1) were upregulated. These results demonstrate that nM concentrations of c-Src kinase inhibitors (1l and PP-1), by reducing the production of VEGF released by tumor cell and its endothelial cell responses, have a highly selective antiangiogenesis effect, which might be useful in combination therapies. PMID- 17131344 TI - Disease-associated casein kinase I delta mutation may promote adenomatous polyps formation via a Wnt/beta-catenin independent mechanism. AB - The Wnt signaling pathway is critical for embryonic development and is dysregulated in multiple cancers. Two closely related isoforms of casein kinase I (CKIdelta and epsilon) are positive regulators of this pathway. We speculated that mutations in the autoinhibitory domain of CKIdelta/epsilon might upregulate CKIdelta/epsilon activity and hence Wnt signaling and increase the risk of adenomatous polyps and colon cancer. Exons encoding the CKIepsilon and CKIdelta regulatory domains were sequenced from DNA obtained from individuals with adenomatous polyps and a family history of colon cancer unaffected by familial adenomatous polyposis or hereditary nonpolyposis colorectal cancer (HNPCC). A CKIdelta missense mutation, changing a highly conserved residue, Arg324, to His (R324H), was found in an individual with large and multiple polyps diagnosed at a relatively young age. Two findings indicate that this mutation is biologically active. First, ectopic ventral expression of CKIdelta(R324H) in Xenopus embryos results in secondary axis formation with an additional distinctive phenotype (altered morphological movements) similar to that seen with unregulated CKIepsilon. Second, CKIdelta(R324H) is more potent than wildtype CKIdelta in transformation of RKO colon cancer cells. Although the R324H mutation does not significantly change CKIdelta kinase activity in an in vitro kinase assay or Wnt/beta-catenin signal transduction as assessed by a beta-catenin reporter assay, it alters morphogenetic movements via a beta-catenin-independent mechanism in early Xenopus development. This novel human CKIdelta mutation may alter the physiological role and enhance the transforming ability of CKIdelta through a Wnt/beta-catenin independent mechanism and thereby influence colonic adenoma development. PMID- 17131345 TI - Polymorphisms in the genes ERCC2, XRCC3 and CD3EAP influence treatment outcome in multiple myeloma patients undergoing autologous bone marrow transplantation. AB - Individual variations in the ability to cope with DNA damage by DNA repair may be essential for the response to chemotherapy, since cancer cells from patients with an effective DNA repair may survive treatment. We have studied the effect on time to treatment failure (TTF) and overall survival (OS) of polymorphism in the DNA repair genes ERCC1, ERCC2 and XRCC3, and in the apoptotic genes PPP1R13L and CD3EAP in 348 patients with multiple myeloma undergoing autologous bone marrow transplantation. Carriers of the variant C-allele of ERCC2 K751Q, the variant T allele of XRCC3 T241M and the variant A-allele of CD3EAP G-21A had a 1.3-fold, 1.8-fold and 1.9-fold longer TTF, respectively, than homozygous wild type carriers (p = 0.006, p = 0.004, p < 0.001). The polymorphism CD3EAP G-21A also had significant effect on OS (p < 0.045). The polymorphism ERCC2 K751Q may to be related to sex, since the prolonged TTF was only seen in women (p = 0.001). Carriers of the combination of variant alleles of ERCC2 K751Q and XRCC3 T241M had 2.8-fold longer TTF (p = 0.0002). This indicates that suboptimal repair of both DNA mechanisms favors prolonged TTF and that polymorphism in ERCC2, XRCC3 and CD3EAP predicts the outcome for patients treated with autologous stem cell transplantation. PMID- 17131346 TI - Imatinib mesylate potentiates topotecan antitumor activity in rhabdomyosarcoma preclinical models. AB - High levels of PDGFR expression in primary rhabdomyosarcoma (RMS) have been associated with disease progression. To date however, there are no reports on the activity of imatinib mesylate, a selective PDGFR inhibitor, in RMS preclinical models. A panel of 5 RMS cell lines was used to investigate the expression of PDGFRalpha and PDGFRbeta, c-Kit and the multidrug transporter ABCG2 (also inhibited by imatinib). In vitro and in vivo experiments were performed using RD (embryonal) and RH30 (alveolar) cell lines to determine the efficacy of imatinib as single agent and in combination with topotecan (TPT). PDGFRbeta was significantly expressed in all cell lines, with the highest levels in RD, while PDGFR alpha and ABCG2 were significantly expressed only in RH30 and RMZ-RC2. c Kit was not detected. PDGFRbeta signaling was active in RD but not in RH30, whilst PDGFRalpha signaling was not active in either cell lines. Significant ABCG2-mediated extrusion of Hoechst 33342 was demonstrated in RH30 but not in RD, and was inhibited by imatinib and the specific ABCG2 inhibitor Ko143. In vitro, imatinib was not active as a single agent at therapeutic concentrations, but significantly potentiated TPT antitumor activity in both cell lines. In vivo experiments using tumor xenografts confirmed the synergistic interaction in both cell lines. These results suggest that at least 2 different mechanisms- inhibition of ABCG2 and/or PDGFRbeta--are involved in the synergistic interaction between imatinib and TPT, and support the use of this combination for the treatment of high-risk RMS patients. PMID- 17131347 TI - Micronutrients and the risk of renal cell cancer: a case-control study from Italy. AB - The role of various micronutrients on the risk of renal cell cancer (RCC) was examined in a multicentric case-control study from Italy, in which information on dietary habits were collected using a validated food-frequency questionnaire. Cases were 767 patients (494 men and 273 women) with incident, histologically confirmed RCC; controls were 1,534 subjects (988 men and 546 women) admitted to the same hospitals as cases for a wide spectrum of acute, nonneoplastic conditions. After allowing for energy and other major covariates, a significant inverse association was found for vitamin E (odds ratio, OR, for the highest quintile of intake versus the lowest one 0.56, 95% confidence interval, CI 0.41 0.75), and vitamin C (OR = 0.72, 95% CI = 0.54-0.96), although the trend in risk for vitamin C was of borderline significance. No significant trend of decreasing risk was found for other micronutrients analyzed, although for most of them the risk estimates were below unity for intakes above the lowest. The ORs for the upper quintile of intake when compared with the lowest one were 0.80 (95% confidence interval, CI = 0.59-1.08) for retinol, 0.82 (95% CI = 0.61-1.10) for alpha-carotene, 0.90 (95% CI = 0.68-1.20) for beta-carotene, 0.94 (95% CI = 0.73 1.21) for beta-criptoxanthin, 0.85 (95% CI = 0.63-1.14) for lutein/zeaxanthin, 0.76 (95% CI = 0.57-1.01) for vitamin D, 0.75 (95% CI = 0.55-1.01) for thiamine, 0.88 (95% CI = 0.66-1.19) for riboflavin, 0.85 for vitamin B6 (95% CI = 0.64 1.13), 0.85 (95% CI = 0.64-1.12) for folate and 0.80 (95% CI = 0.60-1.07) for niacin. No meaningful associations emerged for lycopene (OR = 1.11). The present findings support a possible beneficial effect of vitamin E and C on RCC. PMID- 17131348 TI - Effects of prayer and religious expression within computer support groups on women with breast cancer. AB - Research indicates that two common ways breast cancer patients or women with breast cancer cope with their diagnosis and subsequent treatments are participating in computer support groups and turning to religion. This study is the first we are aware of to examine how prayer and religious expression within computer support groups can contribute to improved psychosocial outcomes for this population. Surveys were administered before group access and then 4 months later. Message transcripts were analyzed using a word counting program that noted the percentage of words related to religious expression. Finally, messages were qualitatively reviewed to better understand results generated from the word counting program. As hypothesized, writing a higher percentage of religion words was associated with lower levels of negative emotions and higher levels of health self-efficacy and functional well-being, after controlling for patients' levels of religious beliefs. Given the proposed mechanisms for how these benefits occurred and a review of the support group transcripts, it appeared that several different religious coping methods were used such as putting trust in God about the course of their illness, believing in an afterlife and therefore being less afraid of death, finding blessings in their lives and appraising their cancer experience in a more constructive religious light. PMID- 17131349 TI - Smaller regional volumes of brain gray and white matter demonstrated in breast cancer survivors exposed to adjuvant chemotherapy. AB - BACKGROUND: Previous studies have shown cognitive impairment in breast cancer survivors who were exposed to adjuvant chemotherapy. Neural damage by chemotherapy might have played some part in these findings. The current study explored the regional brain volume difference between breast cancer survivors exposed to adjuvant chemotherapy (C+) and those unexposed (C-). METHODS: High resolution 1.5-tesla brain magnetic resonance imaging (MRI) databases of breast cancer survivors and healthy controls were used. Brain images were preprocessed for optimal voxel-based morphometry. Comparisons of gray matter and white matter were performed between the C+ and the C- groups, by using MRI scans from within 1 year (the 1-year study, n = 51 and n = 55, respectively) or 3 years after their cancer surgery (the 3-year study, n = 73 and n = 59, respectively). As exploratory analyses, correlation analyses were performed between indices of the Wechsler Memory Scale-Revised and regional brain volume where the volume were significantly smaller. As a reference, MRI scans of cancer survivors were compared with those of healthy controls (n = 55 for the 1-year study and n = 37 for the 3-year study). RESULTS: The C+ patients had smaller gray matter and white matter including prefrontal, parahippocampal, and cingulate gyrus, and precuneus in the 1-year study. However, no difference was observed in the 3-year study. The volumes of the prefrontal, parahippocampal gyrus, and precuneus were significantly correlated with indices of attention/concentration and/or visual memory. Comparisons with healthy controls did not show any significant differences. CONCLUSIONS: Adjuvant chemotherapy might have an influence on brain structure, which may account for previously observed cognitive impairments. PMID- 17131350 TI - Second solid cancers after allogeneic hematopoietic stem cell transplantation. AB - BACKGROUND: The objective of this study was to establish the incidence and risk factors for the development of second solid cancers after allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: The authors reviewed the case files of 926 consecutive patients who underwent allo-HSCT at their institution between 1985 and 2003. RESULTS: Twenty-eight patients developed 30 solid malignancies at a median of 6.8 years after allo-HSCT (range, 0.12-17.3 years) for a 10-year cumulative incidence of 3.1% (95% confidence interval [95% CI], 2-5%; all solid tumors) and 2.3% (95% CI 1-4%; excluding basal cell carcinoma and carcinoma in situ). The risk ratio of developing a second solid malignancy after allografting, compared with the general population of British Columbia adjusted for age and sex, was 1.85 (95% CI, 1.04-3.06; P = .019). In multivariate analysis, recipient age at allo-HSCT >40 years (P = .005) and having a woman donor (P = .0008) were associated with a greater risk of developing a second solid cancer. CONCLUSIONS: The authors concluded that patients undergoing allografting are at increased risk of developing a second solid cancer compared with the general population, particularly those of advanced age at the time of allograft. It is noteworthy that patients who had women as graft donors had an increased risk for developing a second solid cancer. This unexpected finding is a new observation and has not been reported previously. Extended follow-up will be needed to assess more fully the incidence and risk factors for the development of solid cancers, because the latency can be prolonged. PMID- 17131351 TI - Psychobiological effects observed in obese men experiencing body weight loss plateau. AB - Our objective was to investigate the psychobiological impact associated with resistance to further weight loss in obese men. Anthropometric and body composition measurements, resting metabolic rate (RMR) measurement, appetite sensation markers, and three questionnaires [Short Form-36 Health Survey (SF-36), Three-Factor Eating Questionnaire (TFEQ), and Beck Depression Inventory (BDI)] were assessed at baseline and after 1 month of body weight loss plateau induced by a supervised diet and exercise clinical intervention in 11 obese men. The weight loss plateau corresponded to 11.2% of initial body weight (93.9% from fat stores). However, this amount of weight loss induced a significant decrease in RMR (P <.05) and a significant increase in hunger (P <.05) and desire to eat (P <.05). Using the SF-36 Health Survey, physical and mental health were shown to be unchanged at plateau as compared to baseline. The TFEQ showed that cognitive dietary restraint increased (P <.001) compared to baseline. Finally, depression risk as measured by the BDI significantly increased at plateau (P <.01) compared to baseline. Weight loss until resistance to further weight loss may be detrimental for some psychobiological variables including depression, which emphasizes the relevance of caution and reasonable objectives when prescribing a weight reduction program for obese individuals. PMID- 17131352 TI - Longitudinal assessment of symptom and subtype categories in obsessive-compulsive disorder. AB - Although it has been postulated that symptom subtypes are potential predictors of treatment response, few data exist on the longitudinal course of symptom and subtype categories in obsessive-compulsive disorder (OCD). Putative subtypes of OCD have gradually gained more recognition, but as yet there is no generally accepted subtype discrimination. Subtypes, it has been suggested, could perhaps be discriminated based on autogenous versus reactive obsessions stemming from different cognitive processes. In this study, our aim was to assess whether symptom and subtype categories change over time. Using the Yale-Brown Obsessive Compulsive Symptom Checklist (Y-BOCS-SC), we assessed 109 patients who met DSM-IV criteria for OCD to establish baseline values, then reassessed 91 (83%) of the initial group after 36+/-8.2 months. Upon reassessment, we found significant changes from baseline within aggressive, contamination, religious, symmetry and miscellaneous obsessions and within checking, washing, repeating, counting and ordering compulsion categories. Sexual, hoarding, and somatic obsessions, and hoarding and miscellaneous compulsions, did not change significantly. In accordance with the relevant literature, we also assigned patients to one of three subtypes--autogenous, reactive, or mixed groups. Though some changes in subtype categories were found, no subtype shifts (e.g., autogenous to reactive or reactive to autogenous) were observed during the course of the study. Significantly more patients in the autogenous group did not meet OCD criteria at follow-up than did patients in the other groups. Our results suggest that the discrimination between these two types of obsession might be highly valid, because autogenous and reactive obsessions are quite different, both in the development and maintenance of their cognitive mechanisms, and in their outcome. PMID- 17131353 TI - Patients with a major depressive episode responding to treatment with repetitive transcranial magnetic stimulation (rTMS) are resistant to the effects of rapid tryptophan depletion. AB - Repetitive transcranial magnetic stimulation (rTMS) appears to be efficacious in the treatment of major depression based on the results of controlled studies, but little is known about its antidepressant mechanism of action. Mood sensitivity following rapid tryptophan depletion (RTD) has been demonstrated in depressed patients responding to SSRI antidepressants and phototherapy, but not in responders to electroconvulsive therapy (ECT). We sought to study the effects of RTD in patients with major depression responding to a course of treatment with rTMS. Twelve subjects treated successfully with rTMS monotherapy underwent both RTD and sham depletion in a double-blind crossover design. Depressive symptoms were assessed using both a modified Hamilton Depression Rating Scale (HDRS) and Beck Depression Inventory (BDI). The differential change in depression scores across the procedures was compared. No significant difference in mood symptoms was noted between RTD and the sham-depletion procedure on either continuous measures of depression, or in the proportions of subjects that met predefined criteria for a significant degree of mood worsening. Responders to rTMS are resistant to the mood perturbing effects of RTD. This suggests that rTMS does not depend on the central availability of serotonin to exert antidepressant effects in major depression. PMID- 17131355 TI - 2,2'-thiodiethanol: a new water soluble mounting medium for high resolution optical microscopy. AB - The use of high numerical aperture immersion lenses in optical microscopy is compromised by spherical aberrations induced by the refractive index mismatch between the immersion system and the embedding medium of the sample. Especially when imaging >10 micro m deep into the specimen, the refractive index mismatch results in a noticeable loss of image brightness and resolution. A solution to this problem is to adapt the index of the embedding medium to that of the immersion system. Unfortunately, not many mounting media are known that are both index tunable as well as compatible with fluorescence imaging. Here we introduce a nontoxic embedding medium, 2,2'-thiodiethanol (TDE), which, by being miscible with water at any ratio, allows fine adjustment of the average refractive index of the sample ranging from that of water (1.33) to that of immersion oil (1.52). TDE thus enables high resolution imaging deep inside fixed specimens with objective lenses of the highest available aperture angles and has the potential to render glycerol embedding redundant. The refractive index changes due to larger cellular structures, such as nuclei, are largely compensated. Additionally, as an antioxidant, TDE preserves the fluorescence quantum yield of most of the fluorophores. We present the optical and chemical properties of this new medium as well as its application to a variety of differently stained cells and cellular substructures. PMID- 17131356 TI - Quantization of widefield fluorescence images using structured illumination and image analysis software. AB - It is difficult to obtain precise quantitative measurements from fluorescent images captured from widefield microscopes. We wished to ascertain if reliable quantitative measurements of both biological and nonbiological specimens were possible using a widefield microscope equipped with a structured illumination system and image analysis software. In a nonbiological specimen, images were obtained from fluorescent beads of known intensity. For a biologically relevant model we used the uptake of fluorescently labeled transferrin by HeLa cells in culture. In the bead sample, the mean intensity of beads acquired with the structured illumination system showed relative intensities near to the predicted values without any further manipulation of data. In the transferrin uptake experiments, uptake and subsequent recycling of the labeled transferrin could be accurately and reproducibly measured. We conclude that using a combination of structured illumination and image processing algorithms accurate quantization of fluorescent images can be achieved in widefield microscopy. PMID- 17131357 TI - Monitoring of the heavy-metal hyperaccumulation in vegetal tissues by X-ray radiography and by femto-second laser induced breakdown spectroscopy. AB - This article reports on the utilization of X-ray microradiography and laser induced breakdown spectroscopy (LIBS) techniques for investigation of the metal accumulation in different part of leaf samples. The potential of the LIBS analysis for finding the proper plant species for phytoremediation is compared with the results of microradiography measurements at the HERCULES source at ENEA, Rome (Italy) and X-ray microradiography experiments at the ELETTRA Synchrotron, Trieste (Italy). PMID- 17131358 TI - X-ray microanalysis of biological material in the frozen-hydrated state by PIXE. AB - Elemental microanalysis of biological material in the frozen-hydrated state using in-vacuum proton induced X-ray emission is described for the first time. For this purpose, a commercially available cryotransfer system was modified and coupled to the experimental chamber of the nuclear microprobe (NMP). The analyzed material was frozen in propane cooled by liquid nitrogen, fractured, carbon coated, and transferred onto the cold stage (100 K) of the nuclear microprobe chamber. Micro PIXE and simultaneous proton backscattering was performed using a 3 MeV proton beam. Quantitative results were obtained by the standardless method, and tested using 20% gelatin standards. Monitoring of the gas composition inside the system by means of mass spectrometry performed before, during, and after proton bombardment showed good stability of the analyzed material for proton currents not exceeding 150 pA. Average concentrations of light elements (C, N, O, and indirectly H) were also obtained by the proton backscattering technique. No losses of elements measurable by particle-induced X-ray emission (PIXE) during proton irradiation were found during repetitive, short analyses of the same micro areas of gelatin standards. Measurements of thick sections of selected plant and animal material in the frozen-hydrated state-leaf sections of the plant Senecio anomalochrous Hilliard (Asteraceae) and larvae of Chysolina pardalina Fabricius (Chrysomelidae)-showed very good preservation of morphology and elemental distribution. Limits of detection of the order of a few micro g g(-1) were obtained for most elements. PMID- 17131359 TI - Mitochondrial ribosomal protein S36 delays cell cycle progression in association with p53 modification and p21(WAF1/CIP1) expression. AB - Ribosomal biogenesis is correlated with cell cycle, cell proliferation, cell growth and tumorigenesis. Some oncogenes and tumor suppressors are involved in regulating the formation of mature ribosome and affecting the ribosomal biogenesis. In previous studies, the mitochondrial ribosomal protein L41 was reported to be involved in cell proliferation regulating through p21(WAF1/CIP1) and p53 pathway. In this report, we have identified a mitochondrial ribosomal protein S36 (mMRPS36), which is localized in the mitochondria, and demonstrated that overexpression of mMRPS36 in cells retards the cell proliferation and delays cell cycle progression. In addition, the mMRPS36 overexpression induces p21(WAF1/CIP1) expression, and regulates the expression and phosphorylation of p53. Our result also indicate that overexpression of mMRPS36 affects the mitochondrial function. These results suggest that mMRPS36 plays an important role in mitochondrial ribosomal biogenesis, which may cause nucleolar stress, thereby leading to cell cycle delay. PMID- 17131361 TI - Sphingosylphosphorylcholine induces apoptosis of endothelial cells through reactive oxygen species-mediated activation of ERK. AB - Sphingosylphosphorylcholine (SPC) produces reactive oxygen species (ROS) in MS1 pancreatic islet endothelial cells. In the present study, we explored the physiological significance of the SPC-induced ROS generation in endothelial cells. SPC induced cell death of MS1 cells at higher than 10 microM concentration through a caspase-3-dependent pathway. SPC treatment induced sustained activation of an extracellular signal-regulated kinase (ERK), in contrast to transient activation of ERK in response to platelet-derived growth factor (PDGF)-BB, which stimulated proliferation of MS1 cells. Both the SPC-induced cell death and ERK activation were abolished by pretreatment of the cells with the MEK inhibitor U0126 or by overexpression of a dominant negative mutant of MEK1 (DN-MEK1). Pretreatment of the cells with N-acetylcysteine, an antioxidant, completely prevented the SPC-induced ROS generation, apoptosis, and ERK activation, whereas the ROS generation was not abrogated by treatment with U0126. Consistent with these results, SPC induced cell death of human umbilical vein endothelial cells (HUVECs) through ROS-mediated activation of ERK. These results suggest that the SPC-induced generation of ROS plays a crucial role in the cell death of endothelial cells through ERK-dependent pathway. PMID- 17131360 TI - Curcumin prevents lipopolysaccharide-induced atrogin-1/MAFbx upregulation and muscle mass loss. AB - Because elevated ubiquitin ligase atrogin-1/MAFbx and MuRF1 mediate skeletal muscle wasting associated with various catabolic conditions, the signaling pathways involved in the upregulation of these genes under pathological conditions are considered therapeutic targets. AKT and NF-kappaB have been previously shown to regulate the expression of atrogin-1/MAFbx or MuRF1, respectively. In addition, we recently found that p38 MAPK mediates TNF-alpha upregulation of atrogin-1/MAFbx expression, suggesting that multiple signaling pathways mediate muscle wasting in inflammatory diseases. To date, however, these advances have not resulted in a practical clinical intervention for disease induced muscle wasting. In the present study, we tested the effect of curcumin--a non-toxic anti-inflammatory reagent that inhibits p38 and NF-kappaB--on lipopolysaccharide (LPS)-induced muscle wasting in mice. Daily intraperitoneal (i.p.) injection of curcumin (10-60 micro g/kg) for 4 days inhibited, in a dose dependent manner, the LPS-stimulated (1 mg/kg, i.p.) increase of atrogin-1/MAFbx expression in gastrocnemius and extensor digitorum longus (EDL) muscles, resulting in the attenuation of muscle protein loss. It should also be noted that curcumin administration did not alter the basal expression of atrogin-1/MAFbx, nor did it affect LPS-stimulated MuRF1 and polyubiquitin expression. LPS activated p38 and NF-kappaB, while inhibiting AKT; whereas, curcumin administration inhibited LPS-stimulated p38 activation, without altering the effect of LPS on NF-kappaB and AKT. These results indicate that curcumin is effective in blocking LPS-induced loss of muscle mass through the inhibition of p38-mediated upregulation of atrogin-1/MAFbx. PMID- 17131362 TI - In vitro and in vivo osteoblastic differentiation of BMP-2- and Runx2-engineered skeletal myoblasts. AB - Genetic engineering with osteogenic factors is a promising approach for cell based therapeutics and orthopedic regeneration. However, the relative efficacy of different strategies for inducing osteoblastic differentiation remains unclear and is further complicated by varied delivery vehicles, cell types, and evaluation criteria. In order to elucidate the effects of distinct gene-based strategies, we quantitatively evaluated osteoblastic differentiation and mineralization of primary skeletal myoblasts overexpressing either the BMP-2 growth factor or Runx2 transcription factor. Retroviral delivery of BMP-2 or Runx2 stimulated differentiation into an osteoblastic phenotype, as demonstrated by the induction of osteogenic gene expression, alkaline phosphatase activity, and matrix mineralization in monolayer culture and on collagen scaffolds both in vitro and in an intramuscular site in vivo. In general, BMP-2 stimulated osteoblastic markers faster and to a greater extent than Runx2, although we also identified experimental conditions under which these two factors produced similar effects. Additionally, Runx2-engineered cells did not utilize paracrine signaling via secreted osteogenic factors, in contrast to cells overexpressing BMP-2, as demonstrated by conditioned media studies and activation of Smad signaling. These results emphasize the complexity of gene therapy-based orthopedic therapeutics as an integrated relationship of differentiation state, construct maturation, and paracrine signaling of osteogenic cells. This study is significant in evaluating proposed therapeutic systems and defining a successful strategy for integrating gene medicine and orthopedic regeneration. PMID- 17131363 TI - Basic fibroblast growth factor (FGF-2): the high molecular weight forms come of age. AB - After over thirty years from its discovery, research on basic fibroblast growth factor (FGF-2) keeps revealing new aspects of the complexity of its gene expression as it evolved in the eukaryotic organisms. The discovery of multiple forms of FGF-2 generated by alternative translation from AUG and non-canonical CUG codons on the same mRNA transcript has led to the characterization of a low molecular weight (LMW) FGF-2 form and various high molecular weight (HMW) forms (four in humans). In this review, we discuss the biochemical features and biological activities of the different FGF-2 forms. In particular, we focus on the properties that are unique to the HMW forms and its biological functions. PMID- 17131364 TI - Ovariectomy-induced bone loss occurs independently of B cells. AB - Estrogen withdrawal is associated with a significant expansion in B cell precursor and mature B cell populations. However, despite significant circumstantial evidence the role of B lineage cells in ovariectomy-induced bone loss in vivo is unclear. In vitro studies have demonstrated that mature B cells have the potential to both positively and negatively impact osteoclastogenesis by virtue of their capacity to secrete pro-osteoclastogenic cytokines including receptor activator of NFkappaB ligand (RANKL), as well as anti-osteoclastogenic cytokines such as osteoprotegerin (OPG) and transforming growth factor beta (TGFbeta). Although several studies have suggested that expansion of the B lineage following ovariectomy may play a key role in the etiology of ovariectomy induced bone loss, in vivo studies to directly test this notion have yet to be conducted. In this study, we performed ovariectomy on microMT(-/-) mice which are specifically deficient in mature B cells. Analysis of bone mineral density (BMD) by dual-energy X-ray absorptiometry (DXA) and micro-computed tomography (CT) demonstrate that mature B cell-deficient mice undergo an identical loss of bone mass relative to wild-type (WT) control mice. Our data demonstrate that mature B cells are not central mediators of ovariectomy-induced bone loss in vivo. PMID- 17131365 TI - Biophysical analysis of prototype microbicidal gels. AB - The objective of this study was to evaluate the distribution and retention (deployment) of four prototype vehicles for delivery of prophylactic microbicides against vaginal HIV transmission. Study gels were created with different molecular compositions, producing different biophysical properties governing vaginal deployment. The study employed three techniques: direct rheological measurement of gel properties, direct observation of gel surface coating erosion, and dissolution by a vaginal fluid simulant, and mathematical modeling of gel squeezing flow processes. Results suggest significant differences in extent of vaginal coating after gel application and in erosion of these gel layers due to contact with ambient vaginal fluid and shearing. The relationships between gel rheological properties, coating flow and erosion of coating were not always anticipated from differences in gel molecular composition. PMID- 17131366 TI - Expression of RBM5-related factors in primary breast tissue. AB - The aim of this study was to examine the expression of the RBM5 tumor suppressor, in relation to RBM6 and RBM10, to obtain a better understanding of the potential role played by these RBM5-related factors in the regulation of RBM5 tumor suppressor activity. Paired non-tumor and tumor samples were obtained from 73 breast cancer patients. RNA and protein expression were examined by semi quantitative reverse transcription-polymerase chain reaction and immunoblot, respectively. Data were analyzed using various statistical methods to test for correlations amongst the RBM5-related factors, and between the factors and various pathological parameters. Most notably, RBM5, RBM10v1, and HER2 protein expression levels were elevated in tumor tissue (P < 0.0001). RBM5 and RBM10v1 protein expression were significantly positively correlated (P < 0.001), as were RBM5 and HER2 protein expression (P < 0.01), in both non-tumor and tumor tissue, whereas RBM10v1 and HER2 protein expression were only marginally correlated, in non-tumor tissue (P < 0.05). Interestingly, RBM5 and RBM10v1 protein expression were both deregulated in relation to RNA expression in tumor tissue. RBM10v2 and RBM6 RNA were highly significantly positively correlated in relation to various factors relating to poor prognosis (P < 0.0001). To our knowledge, this study is the first to examine RBM5 expression at both the RNA and protein level in primary breast tumor tissue, and the first to examine expression of all RBM5-related factors in a comprehensive manner. The results provide a graphic illustration that RBM5-related factors are significantly differentially expressed in breast cancer, and suggest complex inter-related regulatory networks involving alternative splicing, oncogenic expression, and tissue-specific function. PMID- 17131370 TI - Diarylprolinol ethers: expanding the potential of enamine/iminium-ion catalysis. PMID- 17131371 TI - Gold catalysis. AB - Catalysis by gold has rapidly become a hot topic in chemistry, with a new discovery being made almost every week. Gold is equally effective as a heterogeneous or a homogeneous catalyst and in this Review we attempt to marry these two facets to demonstrate this new found and general efficacy of gold. The latest discoveries are placed within a historical context, but the main thrust is to highlight the new catalytic possibilities that gold-catalyzed reactions currently offer the synthetic chemist, in particular in redox reactions and nucleophilic additions to pi systems. Indeed gold has proved to be an effective catalyst for many reactions for which a catalyst had not been previously identified, and many new discoveries are still expected. PMID- 17131373 TI - Incorporation of unnatural amino acids into cytochrome c3 and specific viologen binding to the unnatural amino acid. PMID- 17131376 TI - Impact of silencing HO-2 on EC-SOD and the mitochondrial signaling pathway. AB - The contribution of heme oxygenase HO-2, the primary source of bilirubin and carbon monoxide (CO) under physiological conditions, to the regulation of vascular function has remained largely unexplored. Using siRNA HO-2, we examined the effect of suppressed levels of HO-2 on vascular antioxidant and survival proteins. In vivo HO-2 siRNA treatment decreased the basal levels of EC-SOD, pAKT proteins (serine-473 and threonine-308), without changing Akt protein expression. HO-2 siRNA treatment increased 3-nitrotyrosine (3-NT) and apoptotic signaling kinase-1 (ASK-1) (P < 0.01). HO activity was decreased by the use of siRNA HO-2. We extended these studies to the mitochondria, examining for the presence of HO-1 and its role in the regulation of pro- and anti-apoptotic proteins. HO activity was increased by the administration of CoPP resulting in the translocation of HO 1 into the mitochondria, mainly to the inner face of the mitochondrial inner membrane. These findings suggest that HO-2 is critical in the maintenance of heme homeostasis and also the regulation of apoptosis by controlling levels of EC-SOD, Akt, 3-NT, and ASK-1. In addition, localization of HO-1 in the mitochondrial compartment plays a critical role in mitochondria-mediated apoptosis. PMID- 17131377 TI - Role of ROS and MAPK in TPA-induced ICAM-1 expression in the myeloid ML-1 cell line. AB - Intercellular adhesion molecule 1 (ICAM-1) has been implicated in playing a key role in the mechanism of inflammatory process initiated in response to environmental agents, and during normal hematopoietic cell differentiation. Though induction of ICAM-1 by 12-O-tetradecanoyl-phorbol-13-acetate (TPA) in myeloid cells has been reported, the molecular mechanism by which TPA upregulates ICAM-1 expression remains unclear. In the present study, we investigated the signaling mechanism associated with TPA-induced ICAM-1 expression in ML-1 cells. Herein, our microarray, flow cytometry, and Western blot analysis indicated that ICAM-1 was constitutively expressed at a low level in ML-1 cells, but its expression was further upregulated at both the mRNA and protein levels in response to TPA. ICAM-1 expression in response to TPA was inhibited by pretreatment with GF109203X [a specific inhibitor of protein kinase C (PKC)], or with PD98059 and U0126 (specific inhibitors of MEK), suggesting the importance of PKC, and Erk1/2 signaling cascades in this response. Interestingly, ICAM-1 expression in response to TPA-induced PKC activation was linked to the generation of reactive oxygen species (ROS), as pretreatment with NAC (an ROS scavenger) blocked both ErK1/2 activation and ICAM-1 expression induced by TPA. In addition, TPA-induced ICAM-1 expression was blocked by inhibition of nuclear factor-kappaB (NF-kappaB) activation following pretreatment with BAY11-7085 (a specific inhibitor of NF-kappaB activation). TPA-induced NF-kappaB activation was shown by increased degradation of IkB (NF-kappaB specific inhibitory protein). Together, these observations demonstrated that TPA, a potent activator of PKC, induces ICAM 1 expression via a ROS- and ERK1/2-dependent signaling mechanism in ML-1 cells. PMID- 17131378 TI - The transcription factor KLF11 can induce gamma-globin gene expression in the setting of in vivo adult erythropoiesis. AB - Previous studies in a fetal erythroid cell line demonstrated that the transcription factor, Kruppel-like factor 11 (KLF11), could specifically induce transcription from a gamma-globin gene promoter, and that this induction was mediated through a specific canonical CACCC cis-DNA binding motif. We report here that ectopic expression of KLF11 can also induce fetal gamma-globin gene expression in the setting of adult erythropoiesis both in vitro and in vivo. Studies in an adult-stage murine erythroleukemia (MEL) cell line demonstrated that retrovirus vector-mediated transduction of KLF11 could increase both the amount of expression from a basally active, but not from a overtly silenced, recombinant gamma-globin transgene, as well as the frequency of cells expressing this transgene. A similar pattern of gamma-globin gene induction was also observed both in vitro and in vivo following KLF11 transduction of bone marrow from mice containing a basally active gamma-globin transgene. These studies provide the first evidence that ectopic expression of a transcription factor can induce gamma-globin gene expression in vivo during adult erythropoiesis. PMID- 17131379 TI - Gene structure and seasonal expression of carp fish prolactin short receptor isoforms. AB - The complex adaptive mechanisms that eurythermal fish have evolved in response to the seasonal changes of the environment include the transduction of the physical parameter variations into neuroendocrine signals. Studies in carp (Cyprinus carpio) have indicated that prolactin (PRL) and growth hormone (GH) expression is associated with acclimatization, suggesting that the pituitary gland is a relevant physiological node in this adaptive process. Also, the distinctive pattern of expression that carp prolactin receptor (PRLr) protein depicts upon seasonal acclimatization supports the hypothesis that PRL and its receptor clearly are involved in the new homeostatic stage that the eurythermal fish needs to survive during the cyclical changes of its habitat. Here, we characterize the first prolactin receptor gene in a teleost and show that its expression is not associated with alternative promoters, unlike in humans and rodents. Using the regulatory region to direct the transcription of green fluorescent protein (GFP) in zebrafish embryos, we mapped the appearance of this hormone receptor during fish development. This is the first report identifying a fish prolactin receptor gene expressing transcript isoforms encoding for short forms of the protein (45 kDa). These have been found in osmoregulatory tissues of the carp and are regulated in connection with the seasonal acclimatization of the fish. PMID- 17131380 TI - Suppression effect of seminal vesicle autoantigen on platelet-activating factor induced mouse sperm capacitation. AB - Mammalian sperm gain the ability to fertilize an egg successfully by the capacitation process. An unregulated capacitation process causes sperm to undergo a spontaneous acrosome reaction (AR) and resulting in loss of their fertilization activity. Thus, functional sperm activation is tightly regulated by a capacitation and suppression (decapacitation) mechanism. Factors, such as platelet-activating factor (PAF) present in both sperm and the female genital tract, are able to stimulate sperm capacitation. Seminal plasma is thought to have the ability to suppress sperm capacitation; however, the regulatory mechanisms of seminal plasma protein on sperm capacitation are not well understood. Recently, we demonstrated that seminal vesicle autoantigen (SVA), a major seminal vesicle secretory protein, is able to suppress mouse sperm capacitation. To further study the suppression spectra of SVA on sperm capacitation, we investigated the effect of SVA on PAF-induced mouse sperm capacitation-related signals. Here, we demonstrate that SVA decreases the [Ca(2+)](i) to suppress the PAF's effects on [Ca(2+)](i), the cAMP level, protein tyrosine phosphorylation, and capacitation. The inhibition of PAF-induced protein tyrosine phosphorylation and capacitation by SVA can be reversed by cAMP agonists. Characterization of the interactions of SVA with PAF by TLC overlay and tryptophan fluorescence spectrum analyses indicates that SVA is capable of binding PAF with an apparent dissociation constant K(d) > 50 microM. Together with these results, we demonstrate that SVA deceases [Ca(2+)](i) and cross-talks with PAF-induced intracellular signals to regulate mouse sperm capacitation. PMID- 17131381 TI - GCIP/CCNDBP1, a helix-loop-helix protein, suppresses tumorigenesis. AB - Deletions and/or loss of heterozygosity (LOH) on chromosome 15 (15q15 and 15q21) have been found in several human tumors, including carcinomas of the colorectum, breast, lung, prostate, and bladder, suggesting the presence of potential tumor suppressor gene(s) in this particular region of chromosome 15. GCIP also called CCNDBP1, DIP1, or HHM, localized at chromosome 15q15, is a recently identified helix-loop-helix leucine zipper (HLH-ZIP) protein without a basic region like the Id family of proteins. In this study, we reported that the expression of GCIP was significantly downregulated in several different human tumors, including breast tumor, prostate tumor, and colon tumors. In human colon tumors, both mRNA and protein expression levels of GCIP were decreased significantly compared to the normal tissues. Treatment of colon cancer cells SW480 with sodium butyrate (NaB), which induces colon cancer cell differentiation, can induce the upregulation of GCIP expression, suggesting that the protein functions as a negative regulator in cell proliferation. Overexpression of GCIP in SW480 colon cancer cell line resulted in a significant inhibition on tumor cell colony formation, while silencing of GCIP expression by siRNA can promote cell colony formation. Furthermore, overexpression of GCIP inhibited the transcriptional activity of cyclin D1 promoter and the expression of cyclin D1 protein in the cell. Finally, we demonstrate that GCIP specifically interacts with one of the class III HDAC proteins, SirT6, which is important for maintaining genome stability. Together, our data suggest a possible function of GCIP in tumor suppression. PMID- 17131382 TI - Animal models for chronic lymphocytic leukemia. AB - B-cell chronic lymphocytic leukemia (B-CLL), the most common leukemia in the Western world, results from an expansion of a rare population of CD5+ mature B lymphocytes. Although clinical features and genomic abnormalities in B-CLL have been studied in considerable detail, the molecular mechanisms underlying disease development has remained unclear until recently. In the last 4 years, several transgenic mouse models for B-CLL were generated. Investigations of these mouse models revealed that deregulation of three pathways, Tcl1-Akt pathway, TNF-NF-kB pathway, and Bcl2-mediated anti-apoptotic pathway, result in the development of B CLL. While deregulation of TCL1 alone caused a B-CLL phenotype in mice, overexpression of Bcl2 required aberrantly activated TNF-NF-kB pathway signaling to yield the disease phenotype. In this article, we present what has been learned from mice with B-CLL phenotype and how these mouse models of B-CLL were used to test therapeutic treatments for this common leukemia. PMID- 17131383 TI - Nucleolar localization of phosphatidylinositol 4-kinase PI4K230 in various mammalian cells. AB - BACKGROUND: Previous immunohistochemical investigations could not detect PI4K230, an isoform of mammalian phosphatidylinositol 4-kinases (also called type III alpha), in the nucleus and nucleolus of cells in spite of its predicted nuclear localization signals. METHODS: Immunofluorescent detection of PI4K230 and other PI4K isoforms was performed on formaldehyde (PFA) or ethanol fixed cells and rat brain cryosections. Costaining with nucleolin and the effect of siRNA, Triton X 100, DNase, and RNase treatments were also tested to determine the localization of PI4K230. RESULTS: PI4K230 gives a prominent signal in the nucleolus of ethanol fixed rat brain cryosections and of several cell types in addition to its presence in the nucleus and cytoplasm. The PI4K230 immunoreactivity of the nucleolus is masked in PFA fixed cells, but it can be restored by treatment of PFA fixed cells with hot wet citrate buffer or by washing the cryosections with PBS prior to PFA fixation. Nucleolar PI4K230 occurs in a Triton X-100 resistant complex. Treatment of COS-7 cells with siRNA targeting PI4K230 and permeabilized B50 cells with DNase or RNase results in the loss of PI4K230 signal from the nucleolus. CONCLUSION: These experiments suggest the participation of PI4K230 in a DNase and RNase sensitive complex with a unique localization and function in the nucleolus. PMID- 17131384 TI - ERK1/2-driven and MKP-mediated inhibition of EGF-induced ERK5 signaling in human proximal tubular cells. AB - The MEK1-ERK1/2 signaling pathway has been implicated in the regulation of renal epithelial cell proliferation, epithelial-to-mesenchymal transition and the induction of an invasive cell phenotype. Much less information is available about the MEK5-ERK5 module and its role in renal epithelial cell proliferation and differentiation. In the present study we have investigated the regulation of these two families of extracellular signal-regulated kinases in epidermal growth factor (EGF)-stimulated human kidney-2 (HK-2) cells and a possible interaction between ERK1/2 and ERK5. Here we report that 5 ng/ml EGF led to a strong stimulation of HK-2 cell proliferation, which was largely U0126-sensitive. Both synthetic MEK1/2 inhibitors U0126 and Cl-1040, when used at 10 and 1 microM, respectively, inhibited basal and EGF-induced ERK1/2 phosphorylation but not ERK5 phosphorylation. Long-term inhibition of MEK1/2-ERK1/2 signaling and/or vanadate sensitive protein phosphatases enhanced and prolonged EGF-induced ERK5 phosphorylation, while transient expression of an adenoviral constitutively active MEK1 (Ad-caMEK1) construct completely blocked EGF-induced ERK5 phosphorylation. Expression of Ad-caMEK1 in HK-2 cells resulted in the upregulation of the dual-specificity phosphatases MKP-3/DUSP6, MKP-1/DUSP1, and DUSP5. The EGF-mediated time-dependent induction of MKP-3, MKP-1 and DUSP5 mRNA levels was U0126-sensitive at a concentration, which blocked EGF-mediated ERK1/2 phosphorylation but not ERK5 phosphorylation. Furthermore, U0126 inhibited EGF induced MKP-3 and MKP-1 protein expression. Both MKP-3 and MKP-1 co immunoprecipitated with ERK5 in unstimulated as well as in EGF-stimulated HK-2 cells. These results suggest the existence of an ERK1/2-driven negative feed-back regulation of ERK5 signaling in EGF-stimulated HK-2 cells, which is mediated by MKP-3, DUSP5 and/or MKP-1. PMID- 17131385 TI - Exposure to azide markedly decreases the abundance of mRNAs encoding cholesterol synthetic enzymes and inhibits cholesterol synthesis. AB - This study was performed to identify genes that are regulated in the adaptive response to prolonged inhibition of oxidative phosphorylation. Gene microarray analysis in control Clone 9 cells and Clone 9 cells exposed to 5 mM azide for 24 h was carried out as a condition of "Chemical hypoxia." Among several hundred mRNAs whose abundances were either increased or decreased, we noted that the abundance of mRNAs encoding enzymes that catalyze the sequential steps of cholesterol synthesis was decreased; this finding was verified by real-time PCR. Exposure to azide for 24 h markedly inhibited the biosynthesis of cholesterol by approximately 90% and decreased the cellular content of cholesterol by 30%, similar results were observed in HepG2 cells. The abundance of sterol regulatory element binding protein (SREBP)-2 mRNA decreased to 0.37 and 0.25 that of controls after 2 and 24 h exposure, respectively. After 24 h of exposure to azide the precursor and nuclear forms of SREBP-2 protein decreased by approximately 80% and approximately 50%, respectively. Stimulation of AMP-activated protein kinase (AMPK) by AICAR in Clone 9 cells increased the abundance of mRNAs encoding cholesterol biosynthetic enzymes and that of SREBP-1c, and had no effect on SREBP 2 mRNA abundance. We conclude that the decrease in the abundance of multiple mRNAs encoding cholesterol biosynthetic enzymes may be mediated by decreased expression of SREBP-2 mRNA and protein and does not involve stimulation of AMPK. The decrease in SREBP-2 mRNA and protein abundance in the face of decreased cell cholesterol content raises the possibility of a novel regulatory pathway. PMID- 17131386 TI - Apoptotic signaling in methylglyoxal-treated human osteoblasts involves oxidative stress, c-Jun N-terminal kinase, caspase-3, and p21-activated kinase 2. AB - Methylglyoxal (MG) is a reactive dicarbonyl compound endogenously produced mainly from glycolytic intermediates. MG is cytotoxic through induction of cell death, and elevated MG levels in diabetes patients are believed to contribute to diabetic complications. In this report, we show for the first time that MG treatment triggers apoptosis in human osteoblasts. We further show that MG induced apoptosis of osteoblasts involves specific apoptotic biochemical changes, including oxidative stress, c-Jun N-terminal kinase (JNK) activation, mitochondrial membrane potential changes, cytochrome C release, increased Bax/Bcl 2 protein ratios, and activation of caspases (caspase-9, caspase-3) and p21 activated protein kinase 2 (PAK2). Treatment of osteoblasts with SP600125, a JNK specific inhibitor, led to a reduction in MG-induced apoptosis and decreased activation of caspase-3 and PAK2, indicating that JNK activity is upstream of these events. Experiments using anti-sense oligonucleotides against PAK2 further showed that PAK2 activation is required for MG-induced apoptosis in osteoblasts. Interestingly, we also found that MG treatment triggered nuclear translocation of NF-kappaB, although the precise regulatory role of NF-kappaB activation in MG induced apoptosis remains unclear. Lastly, we examined the effect of MG on osteoblasts in vivo, and found that exposure of rats to dietary water containing 100-200 microM MG caused bone mineral density (BMD) loss. Collectively, these results reveal for the first time that MG treatment triggers apoptosis in osteoblasts via specific apoptotic signaling, and causes BMD loss in vivo. PMID- 17131387 TI - Trisomy 12 in HESC leads to no selective in vivo growth advantage in teratomas, but induces an increased abundance of renal development. AB - The aim of this investigation was to examine the impact of chromosome 12 amplification (tri-12 cells) in human embryonic stem cells (HESC), following in vivo engraftment to an immunodeficient xeno-model. For this we used sublines from the HESC line HS181, spontaneously exhibiting either low or high frequencies of tri-12 cells. Fluorescent in situ hybridization (FISH) analysis revealed a random distribution of tri-12 cells in the HS181 colonies in vitro. Similarly, the contribution of tri-12 cells to the development of various tissues in teratomas in vivo seemed to be fully random with no particular preference regarding in vivo differentiation pathway of tri-12 HS181 cells compared to HS181 cells with disomy 12 (di-12 cells). On the other hand, following in vivo transplantation the ratio of tri-12/di-12 cells was significantly reduced (P < 0.001), indicating a negative selection for this trisomy in vivo. Moreover, injection of HS181 cultures containing tri-12 cells resulted in a significantly increased abundance of areas compatible with renal formation (P < 0.001), relative teratomas derived from injection of di-12 HS181 cells. However, such areas included no increased relative frequency of tri-12 cells, suggesting indirect mechanism(s) for the increased abundance of renal development. The reasons for such developmental bias are unknown and warrant further investigation. PMID- 17131388 TI - Ubiquitin-dependent proteolysis of trihydrophobin 1 (TH1) by the human papilloma virus E6-associated protein (E6-AP). AB - Human Papilloma virus E6-associated protein (E6-AP), which is known as an E3 ubiquitin ligase, mediates ubiquitination and subsequent degradation of a series of cellular proteins. In this paper, we identify here trihydrophobin 1 (TH1), an integral subunit of the human negative transcription elongation factor (NELF) complex, as a novel E6-AP interaction protein and a target of E6-AP-mediated degradation. Overexpression of E6-AP results in degradation of TH1 in a dose dependent manner, whereas knock-down of endogenous E6-AP elevates the TH1 protein level. TH1 protein turnover is substantially faster, compared to controls, in cells that overexpressed E6-AP. Wild-type E6-AP promotes the ubiquitination of TH1, while a catalytically inactive point mutant of E6-AP abolishes its ubiquitination. Furthermore, in vitro ubiquitination assay also demonstrates that TH1 can be ubiquitinated by E6-AP. The degradation is blocked by treatment with proteasome inhibitor MG132. Herein, we provide strong evidence that TH1 is a specific substrate that is targeted for degradation through E6-AP-catalyzed polyubiquitination. PMID- 17131389 TI - Oxidative stress in the aging murine olfactory bulb: redox proteomics and cellular localization. AB - A recent proteomics analysis from our laboratory demonstrated that several oxidative stress response proteins showed significant changes in steady-state levels in olfactory bulbs (OBs) of 20- vs. 1.5-month-old mice. Oxidative stress may result in protein oxidation. In this study, we investigated two forms of protein oxidative modification in murine OBs: carbonylation and nitration. Redox proteomics with two-dimensional gel electrophoresis, Western blotting, protein digestion, and mass spectrometry was used to quantify total and specific protein carbonylation and to identify differentially carbonylated proteins and determine the carbonylation status of previously identified proteins in OBs of 1.5- and 20 month-old mice. Immunohistochemistry was used to demonstrate the relative intensity and localization of protein nitration in OBs of 1.5-, 6-, and 20-month old mice. Total protein carbonylation was significantly greater in OBs of 20- vs. 1.5-month-old mice. Aldolase 1 (ALDO1) showed significantly more carbonylation in OBs from 20- vs. 1.5-month-old mice; heat shock protein 9A and dihydropyrimidinase-like 2 showed significantly less. Several previously investigated proteins were also carbonylated, including ferritin heavy chain (FTH). Nitration, identified by 3-nitrotyrosine immunoreactivity, was least abundant at 1.5 months, intermediate at 6 months, and greatest at 20 months and was localized primarily in blood vessels. Proteins that were specific targets of oxidation were also localized: ALDO1 in astrocytes of the granule cell layer and FTH in mitral/tufted cells. These results indicate that specific carbonylated proteins, including those in astrocytes and mitral/tufted neurons, and nitrated proteins in the vasculature are molecular substrates of age-related olfactory dysfunction. PMID- 17131390 TI - Manipulation of proliferation and differentiation of human bone marrow-derived neural stem cells in vitro and in vivo. AB - Recent evidence has demonstrated that neural stem cells (NSC) can be expanded from a variety of sources, including embryos, fetuses, and adult bone marrow and brain tissue. We have previously reported the generation of adult rat bone marrow derived cellular spheres that are morphologically and phenotypically similar to neurospheres derived from brain NSC. Here we show that adult human bone marrow derived neural stem cells (HBM-NSC) are capable of generating spheres that are similar to brain neural-derived neurospheres. Additionally, we sought to promote proliferation and differentiation of HBM-NSC through transduction with nonreplicative recombinant adenovirus encoding the cDNA sequence for Gli, rADV Gli-1; sonic hedgehog, rADV-Shh; or Nurr1, rADV-Nurr1. Immunocytochemistry and RT PCR analysis showed that HBM-NSC could be efficiently expanded and differentiated in vitro and that HBM-NSC transduced with rADV-Gli-1 or rADV-Shh dramatically increased NSC time-related proliferation; however, Nurr1 had no effect on proliferation. We also transplanted HBM-NSC into chicken embryos to examine their potential function in vivo. We found that transduction of HBM-NSC with rADV-Gli-1 or rADV-Shh and subsequent transplantation into chicken embryos increased HBM-NSC proliferation, whereas rADV-Nurr1 promoted migration and differentiation in vivo. Our findings suggest that HBM-NSC can be efficiently expanded and differentiated in vitro and in vivo by overexpressing Gli-1, Shh or Nurr1. PMID- 17131391 TI - Abeta(25-35) and its C- and/or N-blocked derivatives: copper driven structural features and neurotoxicity. AB - The toxic properties of beta-amyloid protein, Abeta(1-42), the major component of senile plaques in Alzheimer's disease, depend on nucleation-dependent oligomerization and aggregation. In addition, Abeta(1-42) toxicity is favored by the presence of trace metals, which affect the secondary structure of the peptide. A peptide comprising 11 residues within Abeta(1-42) [Abeta(25-35)] aggregates and retains the neurotoxic activity of Abeta(1-42). We have used both Abeta(25-35) and its C-amidated or N-acetylated/C-amidated derivatives to investigate the role of copper(II) in modulating the conformation and aggregation state as well as the neurotoxic properties of amyloid peptides. Electrospray ionization mass spectrometry (ESI-MS) and electron paramagnetic resonance (EPR) measurements were performed to verify the formation of copper(II)/Abeta(25-35) complexes and to determine the coordination mode, respectively. Abeta(25-35) and its derivatives were analyzed by circular dichroism spectroscopy to assess their secondary structure, subjected to thioflavine-T (Th-T) binding assay to reveal beta-sheet structured aggregates formation, and imaged by scanning force microscopy. Toxicity was assessed on mature cultures of rat cortical neurons. We found that beta-sheet-structured species of Abeta(25-35) were neurotoxic, whereas the random-coil-structured derivatives were devoid of effect. Interestingly, copper promoted the random-coil/beta-sheet transition of Abeta(25-35), with ensuing peptide toxicity, but it induced the toxicity of the N-acetylated/C amidated derivative without affecting peptide folding. Moreover, copper did not influence either the folding or the activity of the C-amidated Abeta(25-35), suggesting that blockade of the C-terminus of Abeta peptides might be sufficient to prevent Abeta toxicity. PMID- 17131392 TI - Changes of amino acid concentrations in the rat vestibular nuclei after inferior cerebellar peduncle transection. AB - Although there is a close relationship between the vestibular nuclear complex (VNC) and the cerebellum, little is known about the contribution of cerebellar inputs to amino acid neurotransmission in the VNC. Microdissection of freeze dried brain sections and high-performance liquid chromatography (HPLC) were combined to measure changes of amino acid concentrations within the VNC of rats following transection of the cerebellovestibular connections in the inferior cerebellar peduncle. Distributions of 12 amino acids within the VNC at 2, 4, 7, and 30 days after surgery were compared with those for control and sham-lesioned rats. Concentrations of gamma-aminobutyric acid (GABA) decreased by 2 days after unilateral peduncle transection in nearly all VNC regions on the lesioned side and to lesser extents on the unlesioned side and showed partial recovery up to 30 days postsurgery. Asymmetries between the two sides of the VNC were maintained through 30 days. Glutamate concentrations were reduced bilaterally in virtually all regions of the VNC by 2 days and showed complete recovery in most VNC regions by 30 days. Glutamine concentrations increased, starting 2 days after surgery, especially on the lesioned side, so that there was asymmetry generally opposite that of glutamate. Concentrations of taurine, aspartate, and glycine also underwent partially reversible changes after peduncle transection. The results suggest that GABA and glutamate are prominent neurotransmitters in bilateral projections from the cerebellum to the VNC and that amino acid metabolism in the VNC is strongly influenced by its cerebellar connections. PMID- 17131393 TI - Total thyroidectomy for differentiated thyroid cancer. AB - There has been a long debate about the optimal surgical management of differentiated thyroid cancer. It has focused on the extent of thyroidectomy, with recommendations ranging from thyroid lobectomy to total thyroidectomy. There is no randomized prospective trial addressing this issue; such a trial would be prohibitive, since differentiated thyroid cancer has a good prognosis and a long natural history. Instead, there is heavy reliance on retrospective analyses, as well as consensus expert opinion and experience. We review this evidence, along with recent recommendations from several professional associations. We believe that total or near-total thyroidectomy followed by (131)I ablation and thyroid hormone suppression are most appropriate for the majority of patients with differentiated thyroid cancer, as retrospective analyses have shown that they reduce the risk of cancer recurrence, address the chance of multifocal intrathyroidal cancer, and facilitate use of surveillance scans and thyroglobulin monitoring for post-operative recurrence. This recommendation comes with the caveat that total thyroidectomy must be performed safely, since there is evidence that surgeon volume is associated with patient outcomes. PMID- 17131394 TI - Seminars: local and regional anesthesia for thyroid surgery. AB - BACKGROUND: Thyroid surgery is performed by a large number of surgeons with varying experience in thyroidectomy. The standard technique involves the use of general anesthesia, which provides patient comfort and virtually unlimited time to conduct the operation. Historically, thyroid surgery was conducted under local anesthesia by surgeons with significant expertise in the treatment of thyroid diseases. Over the past decade, there has been a renewed interest in the art of performing thyroidectomy under local/regional anesthesia in some specialized high volume endocrine surgery centers. METHODS: Here we review the indications and contraindications and technical considerations for performing thyroidectomy under local or regional anesthesia. RESULTS AND CONCLUSION: Local and regional anesthesia is safe and well tolerated for the majority of thyroid surgery. PMID- 17131395 TI - Re-operative neck surgery for well-differentiated thyroid cancer of follicular origin. AB - This review focuses on the pathologic criteria for completion thyroidectomy in well differentiated thyroid cancer as well the diagnosis and treatment of recurrent disease. The roles of ultrasound in the diagnosis of a cervical recurrence, its value in determining the extent of lymph node dissection in the lateral compartment, and the importance of intra-operative ultrasound in re operative thyroid surgery are discussed. PMID- 17131396 TI - Inherited aspects of papillary thyroid carcinoma. AB - The presence of papillary carcinoma of the thyroid in multiple generations of one kindred is a statistical impossibility as an occurrence of chance. However, traditional and molecular genetic analyses to date have failed to support the notion of a single gene mutation or identify one, in distinct contrast to medullary carcinoma of the thyroid. Findings to date, outside of distinct multicancer syndromes, suggest the interplay of inherited susceptibility and other factors, such as environmental exposures. It is possible that the main identifiable genetic risk factors at this time are the presence of multinodular goiter or Hashimoto's thyroiditis within the family. PMID- 17131397 TI - Anaplastic thyroid cancer and primary thyroid lymphoma: a review of these rare thyroid malignancies. AB - BACKGROUND: To review the current literature on the treatment of anaplastic thyroid cancer (ATC) and thyroid lymphoma (TL). RESULTS: Both anaplastic carcinoma (ATC) and TL represent rare forms of thyroid cancer. ATC behaves in a highly aggressive manner, resulting in significant morbidity and mortality. Multimodality therapy consisting of both radiotherapy (RT) and chemotherapy is essential in obtaining local/regional control. Although ATC has been relatively chemo resistant, newer agents such like taxotere show promise. The role of surgery in the treatment of ATC continues to evolve, presently it should be reserved for patients who have shown an initial response to multimodality therapy and in patients in whom a complete macroscopic resection can be achieved with minimal morbidity. The successful treatment of TL currently lies in accurately diagnosing the histological subtype. Both large B-cell and mixed lymphomas are best treated with multimodality therapy consisting of CHOP combined with hyper fractioned RT. MALT lymphomas with there more indolent course may be amenable to single modality RT or total thyroidectomy if diagnosed at an early stage IE. DISCUSSION: Although both ATC and TL are rare, it is important for surgeons to be aware of the need for multimodality therapy when treating these patients and to understand the limited role surgery plays in diagnosis and treatment. PMID- 17131398 TI - Enhanced autophagic cell death in expanded polyhistidine variants of HOXA1 reduces PBX1-coupled transcriptional activity and inhibits neuronal differentiation. AB - HOXA1 is a member of the homeobox gene family and is involved in early brain development. In our previous study, we identified novel variants of polyhistidine repeat tract in HOXA1 gene and showed that ectopic expression of expanded variants led to enhanced intranuclear aggregation and accelerated cell death in a time-dependent manner. Here, we further investigate the implications of polyhistidine variants on HOXA1 function. Aside from intranuclear aggregation, we observed cytosolic aggregates during the early stages of expression. Rapamycin, an autophagy inducer, resulted in decreased protein aggregation and cell death. Here, we also show an interaction between variants of HOXA1 and one of the HOX protein known cofactors, PBX1. Expanded HOXA1 variants exhibited reduced PBX1 coupled transcriptional activity through a regulatory enhancer of HOXB1. Moreover, we demonstrate that both deleted and expanded variants inhibited neurite outgrowth in retinoic acid-induced neuronal differentiation in neuroblastoma cells. These results provide further evidence that expanded polyhistidine repeats in HOXA1 enhance aggregation and cell death, resulting in impaired neuronal differentiation and cooperative binding with PBX1. PMID- 17131399 TI - Compensation by reduced L-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor responses in a mouse model with reduced gamma-aminobutyric acid type A receptor-mediated synaptic inhibition. AB - L-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor antagonists increase the threshold for electroshock-induced convulsions. Here, we show that a transgenic mouse line overexpressing cerebellum-restricted gamma aminobutyric acid type A (GABA(A)) receptor alpha6 subunit in the hippocampal CA1 pyramidal cells (Thy1alpha6 mouse line) exhibits about a 20% increase in the electroshock current intensity inducing tonic hindlimb extension convulsion in 50% of the mice compared with that of their wild-type controls. AMPA receptor mediated miniature excitatory postsynaptic currents (mEPSCs) in patch clamp recordings of CA1 pyramidal neurons in hippocampal slices had decreased amplitudes (8.4 +/- 2.2 pA) in the transgenics compared with the wild types (10.3 +/- 2.5 pA) but showed no change in current decay or frequency. Our results suggest that decreased AMPA-mediated neurotransmission might explain the increased threshold for electroconvulsions and warrant further studies on the regulation between various components of inhibition and excitation in neurons. PMID- 17131400 TI - Calcific myonecrosis and tenosynovitis: sonographic findings with correlative imaging. AB - We report the sonographic appearance of a case of calcific myonecrosis and tenosynovitis with correlative plain radiography and CT findings. Calcific myonecrosis is a rare complication of previous trauma that presents as a tumor like calcified mass years after an initial injury, most commonly affecting the anterior compartment of the lower leg. Although the clinical, radiographic, and MRI features of calcific myonecrosis have been well described, to our knowledge the sonographic appearance has not been reported previously in the literature. PMID- 17131401 TI - Drosophila melanogaster p24 genes have developmental, tissue-specific, and sex specific expression patterns and functions. AB - Genes encoding members of the p24 family of intracellular trafficking proteins are present throughout animal and plant lineages. However, very little is known about p24 developmental, spatial, or sex-specific expression patterns or how localized expression affects function. We investigated these problems in Drosophila melanogaster, which contains nine genes encoding p24 proteins. One of these genes, logjam (loj), is expressed in the adult female nervous system and ovaries and is essential for oviposition. Nervous system-specific expression of loj, but not ovary-specific expression, rescues the behavioral defect of mutants. The Loj protein localizes to punctate structures in the cellular cytoplasm. These structures colocalize with a marker specific to the intermediate compartment and cis-Golgi, consistent with experimental evidence from other systems suggesting that p24 proteins function in intracellular transport between the endoplasmic reticulum and Golgi. Our findings reveal that Drosophila p24 transcripts are developmentally and tissue-specifically expressed. CG31787 is male-specifically expressed gene that is present during the larval, pupal, and adult stages. Female CG9053 mRNA is limited to the head, whereas males express this gene widely. Together, our studies provide experimental evidence indicating that some p24 genes have sex-specific expression patterns and tissue- and sex-limited functions. PMID- 17131402 TI - Preoperative Doppler sonography for prevention of perioperative stroke in head and neck cancer patients undergoing neck dissection: is it beneficial? PMID- 17131403 TI - Temporal and spatial expression profiles of the Fat3 protein, a giant cadherin molecule, during mouse development. AB - Cadherins constitute a superfamily of cell-cell interaction molecules that participate in morphogenetic processes of animal development. Fat cadherins are the largest members of this superfamily, with 34 extracellular cadherin repeats. Classic Fat, identified in Drosophila, is known to regulate cell proliferation and planar cell polarity. Although 4 subtypes of Fat cadherin, Fat1, Fat2, Fat3, and Fat4/Fat-J, have been identified in vertebrates, their protein localization remains largely unknown. Here we describe the mRNA and protein distributions of Fat3 during mouse development. We found that Fat3 expression was restricted to the nervous system. In the brain, Fat3 was expressed in a variety of regions and axon fascicles. However, its strongest expression was observed in the olfactory bulb and retina. Detailed analysis of Fat3 in the developing olfactory bulb revealed that Fat3 mRNA was mainly expressed by mitral cells and that its proteins were densely localized along the dendrites of these cells as well as in their axons to some extent. Fat3 transcripts in the retina were expressed by amacrine and ganglion cells, and its proteins were concentrated in the inner plexiform layer throughout development. Based on these observations, we suggest that Fat3 plays a role in the interactions between neurites derived from specific subsets of neurons during development. PMID- 17131404 TI - Current approaches to medullary thyroid carcinoma, sporadic and familial. AB - Medullary thyroid carcinoma (MTC) is a rare malignancy of the thyroid C cells. It occurs in hereditary (25% of cases) and sporadic forms, and aggressiveness is related to the clinical presentation (hereditary vs. sporadic) and the type of RET mutation present. In hereditary cases, early diagnosis makes preventative surgery possible. In established cases, thorough surgical extirpation of the primary tumor and nodal metastases has been the mainstay of treatment. Radioactive iodine, external beam radiation therapy (EBRT), and conventional chemotherapy have not been effective. Newer systemic treatments, with agents that target abnormal RET proteins, hold promise and are being tested in clinical trials for patients with metastatic disease. PMID- 17131405 TI - Seminars: controversies in the management of pediatric thyroid malignancy. AB - Thyroid cancer in children is a rare malignancy with unusual biological behavior. It often presents at advanced stages, yet behaves in a benign manner, when compared to its adult counterpart. Surgeons have debated the risks and benefits of aggressive surgical procedures for over a decade. A consensus treatment plan for childhood thyroid cancer has not been achieved, however radical surgical procedures have, in general, fallen out of favor. The best operative and adjuvant strategy for these children can only be determined when a better understanding of the tumor cell biology and genetics of this cancer is known. This review discusses the current controversies in the management of differentiated childhood thyroid malignancy. PMID- 17131406 TI - Genetic screen for mutations affecting development and function of the enteric nervous system. AB - An intact enteric nervous system is required for normal gastrointestinal tract function. Several human conditions result from decreased innervation by enteric neurons; however, the genetic basis of enteric nervous system development and function is incompletely understood. In an effort to increase our understanding of the mechanisms underlying enteric nervous system development, we screened mutagenized zebrafish for changes in the number or distribution of enteric neurons. We also established a motility assay and rescreened mutants to learn whether enteric neuron number is correlated with gastrointestinal motility in zebrafish. We describe mutations isolated in our screen that affect enteric neurons specifically, as well as mutations that affect other neural crest derivatives or have pleiotropic effects. We show a correlation between the severity of enteric neuron loss and gastrointestinal motility defects. This screen provides biological tools that serve as the basis for future mechanistic studies. PMID- 17131407 TI - Reduced endothelin converting enzyme-1 and endothelin-3 mRNA in the developing bowel of male mice may increase expressivity and penetrance of Hirschsprung disease-like distal intestinal aganglionosis. AB - Hirschsprung disease (distal intestinal aganglionosis, HSCR) is a multigenic disorder with incomplete penetrance, variable expressivity, and a strong male gender bias. Recent studies demonstrated that these genetic patterns arise because gene interactions determine whether enteric nervous system (ENS) precursors successfully proliferate and migrate into the distal bowel. We now demonstrate that male gender bias in the extent of distal intestinal aganglionosis occurs in mice with Ret dominant-negative mutations (RetDN) that mimic human HSCR. We hypothesized that male gender bias could result from reduced expression of a gene already known to be essential for ENS development. Using quantitative real-time polymerase chain reaction (PCR) we demonstrated reduced levels of endothelin converting enzyme-1 and endothelin-3 mRNA in the male mouse bowel at the time that ENS precursors migrate into the colon. Other HSCR associated genes are expressed at comparable levels in male and female mice. Testosterone and Mullerian inhibiting substance had no deleterious effect on ENS precursor development, but adding EDN3 peptide to E11.5 male RetDN heterozygous mouse gut explants in organ culture significantly increased the rate of ENS precursor migration through the bowel. PMID- 17131408 TI - Seminars in surgical oncology on thyroid cancer. PMID- 17131409 TI - Contemporary diagnostic approach to the thyroid nodule. AB - Thyroid nodules are common, with an estimated incidence of 5%-10% in the United States. The current gold standard for diagnosis is fine needle aspiration biopsy (FNAB). The incidence of indeterminate diagnoses varies from 10% to 25%. Surgical resection is usually indicated to exclude the diagnosis of cancer in these patients. However, only a minority (about 20%) of indeterminate thyroid nodules actually harbor a malignancy, resulting in surgery for diagnostic purposes alone in many patients. The increased detection of benign nodules and microcarcinomas reinforces the need for improved non-operative methods to differentiate benign from malignant disease and discriminate low-risk from high-risk cancers. In this article we present a current, rational diagnostic approach to the patient with a thyroid nodule, evaluate new advances including thyroid genomic and predictor models, and propose the development of prospective trials to incorporate these new additions into clinical decision making. Given how many questions still exist for patients with thyroid nodules, partnership and collaboration, or the "bench to bedside" concept should find its way into most every thyroid surgeon and endocrinologist's lexicon. PMID- 17131410 TI - Surgeon, scientist, and sailor--a tribute to Blake Cady, MD. PMID- 17131411 TI - Pathology and genetics of thyroid carcinoma. AB - Carcinomas of the thyroid comprise a heterogeneous group of neoplasms with distinctive clinical and pathological characteristics. Over the past 15 years, the application of molecular technologies to the study of these neoplasms has elucidated critical genetic pathways associated with the development of specific thyroid tumor types. In papillary thyroid carcinoma (PTC), genetic events involve RET and TRK (rearrangements) and BRAF and RAS (mutations), although RAS mutations are uncommon except in the follicular variant of PTC. These genetic alterations, which rarely overlap in the same tumor, result in signaling abnormalities in the mitogen-activated protein kinase pathway. In contrast, genetic alterations in follicular carcinomas include PAX8-PPARgamma translocations and RAS mutations while mutations of CTNNB1 and p53 have been implicated in the development and progression of poorly differentiated and undifferentiated (anaplastic) thyroid carcinomas. Germline mutations of RET are responsible for the development of heritable forms of medullary thyroid carcinoma (MTC) while somatic mutations of this oncogene are found in a significant proportion of sporadic MTCs. The results of these studies not only have provided additional approaches to thyroid tumor classification, but also have stimulated the development of novel approaches to tumor diagnosis and additional parameters for prognostic assessment and potential biologic therapeutic strategies. PMID- 17131412 TI - In vitro characterization of a human neural progenitor cell coexpressing SSEA4 and CD133. AB - The stage-specific embryonic antigen 4 (SSEA4) is commonly used as a cell surface marker to identify the pluripotent human embryonic stem (ES) cells. Immunohistochemistry on human embryonic central nervous system revealed that SSEA4 is detectable in the early neuroepithelium, and its expression decreases as development proceeds. Flow cytometry analysis of forebrain-derived cells demonstrated that the SSEA4-expressing cells are enriched in the neural stem/progenitor cell fraction (CD133(+)), but are rarely codetected with the neural stem cell (NSC) marker CD15. Using a sphere-forming assay, we showed that both subfractions CD133(+)/SSEA4(+) and CD133(+)/CD15(+) isolated from the embryonic forebrain are enriched in neurosphere-initiating cells. In addition CD133, SSEA4, and CD15 expression is sustained in the expanded neurosphere cells and also mark subfractions of neurosphere-initiating cells. Therefore, we propose that SSEA4 associated with CD133 can be used for both the positive selection and the enrichment of neural stem/progenitor cells from human embryonic forebrain. PMID- 17131413 TI - Chernobyl and thyroid cancer. AB - The most dramatic effect of exposure to fallout from the Chernobyl accident on physical health has been the increase in thyroid cancer. When this was first reported 1,2, there was scepticism in Europe and the US, as it was not thought plausible that exposure to radio-isotopes of iodine in fallout could lead to such an increase in thyroid cancer with such a short latency. This review will discuss the lessons to be learnt from a re-examination of the reasons for the original scepticism, together with what is currently known about the factors that have influenced the development of excess thyroid cancers in those exposed, the number and type of cancers that have occurred, and the interaction between tumor morphology, molecular pathology, clinical behavior, and latent period. PMID- 17131414 TI - Selective surgery and adjuvant therapy based on risk classifications of well differentiated thyroid cancer. AB - BACKGROUND: The prognosis of well-differentiated thyroid cancer has been stratified into low- and high-risk groups. These risk groups can be used to predict prognosis and to guide treatment. METHODS: Retrospective study of 962 patients with well-differentiated thyroid cancer treated from 1940 to 1998. Stratification into low- and high-risk groups based on age, metastases, extent, and size (AMES). Effects on survival of surgery, lymph node dissection, and radiation therapy were examined. RESULTS: Seven hundred twenty-eight cases were papillary and 234 were follicular carcinoma. Seven hundred-fifty cases were low risk and 207 high risk. Twenty-year survival was 97.4% in the low-risk patients and 54.0% in high-risk patients (P < 0.001); it was 63.2% in the younger high risk group and 41.0% in the older high-risk group (P < 0.001). Older high-risk patients had a survival advantage with bilateral thyroidectomy. Extent of surgery did not change survival in either the younger high-risk group or the low-risk group. Lymph node dissection and radioactive iodine ablation did not have an impact on survival. DISCUSSION: Well-differentiated thyroid cancer in low-risk patients has a favorable outcome regardless of treatment. Low-risk patients can be safely treated with unilateral thyroidectomy alone. Risk stratification with a modification of the AMES criteria can be used to guide treatment. PMID- 17131415 TI - Excitotoxic protection by polyanionic polysaccharide: evidence of a cell survival pathway involving AMPA receptor-MAPK Interactions. AB - Growing numbers of studies indicate that polysaccharides influence signaling events important for brain function. It has been speculated that such polysaccharide modulation of neuronal signals can promote synaptogenesis and cell maintenance. Here, we tested whether dextran sulfate, a polyanion that mimics natural mucopolysaccharides, protects hippocampal neurons against excitotoxic insults. An excitotoxin was applied to primary hippocampal cultures in the absence or presence of a large 500-kDa dextran sulfate (DS-L), a smaller 5-8-kDa species (DS-S), or sulfate-free dextran of 500 kDa. Only DS-L prevented neuronal damage as determined by a membrane permeability assay and phase contrast morphology. The sulfate and size dependence is also characteristic of DS-L's modulatory action on the channel activity of alpha-amino-3-hydroxy-5-methyl-4 isoxazole propionic acid (AMPA)-type glutamate receptors. The extent of neuroprotection correlates with the level of modulation of AMPA responses, and DS L exhibits comparable EC(50) values for the two effects (3-7 nM). DS-L also modulates the link between AMPA receptors and mitogen-activated protein kinase (MAPK) involving extracellular signal-regulated protein kinase (ERK), well known for its involvement in cell survival and repair. Correspondingly, protection against N-methyl-D-aspartate (NMDA) excitotoxicity was evident in hippocampal slice cultures when DS-L was applied 30 min postinsult. These findings suggest that polysaccharides elicit neuroprotection in the brain, including enhanced repair responses through the AMPA receptor-MAPK axis. PMID- 17131416 TI - Developmental abnormalities in the nerves of peripheral myelin protein 22 deficient mice. AB - Peripheral myelin protein 22 (PMP22) is a tetraspan glycoprotein whose misexpression is associated with a family of hereditary peripheral neuropathies. In a recent report, we have characterized a novel PMP22-deficient mouse model in which the first two coding exons were replaced by the lacZ reporter. To investigate further the myelin abnormalities in the absence of PMP22, sciatic nerves and dorsal root ganglion (DRG) neuron explant cultures from PMP22 deficient mice were studied at various stages of myelination. Throughout the first 3 months of postnatal development, myelin protein and beta4 integrin levels are dramatically reduced, whereas p75 and beta1 integrin remain elevated. By immunostaining, the distributions of several glial proteins, including beta4 integrin, the voltage-gated potassium channel Kv1.1, and E-cadherin, are altered. Schwann cells from PMP22-deficient mice are able to produce limited amounts of myelin in DRG explant cultures, yet the internodal segments are dramatically fewer and shorter. The comparison of PMP22-deficient mice with other PMP22 mutant models reveals that the decrease in beta4 integrin is specific to an absence of PMP22. Furthermore, whereas lysosome-associated membrane protein 1 and ubiquitin are notably up-regulated in nerves of PMP22-deficient mice, heat shock protein 70 levels remain constant or decrease compared with wild-type or PMP22 mutant samples. Together these results support a role for PMP22 in the early events of peripheral nerve myelination. Additionally, although myelin abnormalities are a commonality among PMP22 neuropathic models, the underlying subcellular mechanisms are distinct and depend on the specific genetic abnormality. PMID- 17131417 TI - Injury-induced Janus kinase/protein kinase C-dependent phosphorylation of growth associated protein 43 and signal transducer and activator of transcription 3 for neurite growth in dorsal root ganglion. AB - Elevation of corticosteroids and excessive glutamate release are the two major stress responses that occur sequentially during traumatic CNS injury. We have previously reported that sequential application of corticosterone and kainic acid (CORT + KA) mimicking the nerve injury condition results in synergistic enhancement of neurite outgrowth and expression of growth-associated protein 43 (GAP-43) in cultured dorsal root ganglion (DRG). GAP-43 is known to promote neurite extension when phosphorylated by protein kinase C (PKC). In addition, PKC can phosphorylate the signal transducer and activator of transcription 3 (STAT3) at Ser727, which is phosphorylated primarily by Janus kinase (JAK) at Tyr705. In this study, we further examine the role of PKC in this stress-induced growth promoting effect. In the cultured DRG neurons, the JAK inhibitor AG-490 and the PKC inhibitor Ro-318220 reduced the CORT + KA-enhanced neurite growth effect when applied prior to CORT and KA treatment, respectively. Both AG-490 and Ro-318220 diminished the CORT + KA-enhanced GAP-43 expression, phosphorylation, and axonal localization. Furthermore, CORT + KA treatment synergistically phosphorylated STAT3 at Ser727 but not at Tyr705. Similar phenomena were observed in an animal model of acute spinal cord injury (SCI), in which phosphorylation of GAP-43 and phospho-Ser727-STAT3 was elevated in the injured DRG 4 hr after the impact injury. Further treatment with the therapeutic glucocorticoid methylprednisolone enhanced the phosphorylation of GAP-43 in both the DRG and the spinal cord of SCI rats. These results suggest that elevated glucocorticoids and overexcitation following CNS injury contribute to nerve regeneration via induction of JAK/PKC mediated GAP-43 and STAT3 activities. PMID- 17131418 TI - Role of p53 in neurotoxicity induced by the endoplasmic reticulum stress agent tunicamycin in organotypic slice cultures of rat spinal cord. AB - The endoplasmic reticulum (ER) is important for maintaining the quality of cellular proteins. Various stimuli can disrupt ER homeostasis and cause the accumulation of unfolded or misfolded proteins, i.e., a state of ER stress. Recently, ER stress has been reported to play an important role in the pathogenesis of neurological disorders such as cerebral ischemia and neurodegenerative diseases, but its involvement in the spinal cord diseases has not been fully discussed. We conducted this study using tunicamycin (Tm) as an ER stress inducer for rat spinal cord in organotypic slice culture, a system that we have recently established. Tm was shown to induce ER stress by increased expression of GRP78. The viability rate of spinal cord neurons decreased in a dose-dependent manner with Tm treatment, and dorsal horn interneurons were more vulnerable to Tm-induced neurotoxicity. A p53 inhibitor significantly increased the viability of dorsal horn interneurons, and immunofluorescence studies showed nuclear accumulation of p53 in the dorsal horns of Tm-treated spinal cord slices. These findings suggest that p53 plays an important role in the killing of dorsal horn interneurons by Tm. In contrast, motor neurons were not protected by the p53 inhibitor, suggesting that the role of p53 may vary between different cell types. This difference might be a clue to the mechanism of the stress-response pathway and might also contribute to the potential application of p53 inhibitors for the treatment of spinal cord diseases, including amyotrophic lateral sclerosis. PMID- 17131419 TI - Androgen regulates trkB immunolabeling in spinal motoneurons. AB - Neurotrophic factors and steroid hormones have been shown to have neuroprotective/neurotherapeutic effects, and it has been shown previously that brain-derived neurotrophic factor (BDNF) and testosterone have a combinatorial effect in the maintenance of motoneurons. Given that gonadal hormones regulate the BDNF receptor, tyrosine receptor kinase B (trkB), we hypothesized that such a regulatory effect could mediate the interactive effects of BDNF and testosterone. Using immunohistochemical methods, we examined the frequency of cells immunolabeled for trkB receptors in two populations of spinal motoneurons, the hormone-sensitive, sexually dimorphic motoneurons of the spinal nucleus of the bulbocavernosus (SNB) and the nondimorphic motoneurons innervating the muscles of the quadriceps. In both the highly androgen-sensitive SNB motoneurons and the more typical somatic motoneurons innervating the quadriceps, the frequency of motoneurons intensely immunolabeled for trkB receptors was regulated by the presence of testosterone. Castrated animals deprived of testosterone showed a reduced frequency of intensely labeled motoneurons compared with intact animals or castrated animals given testosterone replacement. This finding suggests that the combinatorial effect of BDNF and testosterone in the maintenance of motoneurons could occur at least in part through an androgen-mediated expression of the BDNF receptor. PMID- 17131420 TI - Growth and pathfinding of regenerating axons in the optic projection of adult fish. AB - In contrast to mammals, teleost fish are able to regrow severed long-range projection axons in the central nervous system (CNS), leading to recovery of function. The optic projection in teleost fish is used to study neuron-intrinsic and environmental molecular factors that determine successful axon regrowth and navigation through a complex CNS pathway back to original targets. Here we review evidence for regeneration-specific regulation and robust expression of growth- and pathfinding-associated genes in regenerating retinal ganglion cell (RGC) axons of adult fish. The environment of the CNS in fish appears to contain few inhibitory molecules and at the same time a number of promoting molecules for axon regrowth. Finally, some environmental cues that are used as guidance cues for developing RGC axons are also present in continuously growing adult animals. These molecules may serve as guidance cues for the precise navigation of axons from newly generated RGCs in adult animals as well as of regenerating RGC axons after a lesion. The application of new molecular techniques especially to adult zebrafish, is likely to produce new insights into successful axonal regeneration in the CNS of fish and the absence thereof in mammals. PMID- 17131421 TI - RNA interference-mediated knockdown of alpha-synuclein protects human dopaminergic neuroblastoma cells from MPP(+) toxicity and reduces dopamine transport. AB - The critical observation in the pathology of Parkinson's disease (PD) is that neurodegeneration is largely restricted to dopaminergic neurons that develop cytoplasmic inclusions called Lewy bodies. These aggregations contain the protein alpha-synuclein. Furthermore, it is becoming apparent that alpha-synuclein expression levels are a major factor in PD pathogenesis. Patients with additional copies of the alpha-synuclein gene develop PD with a severity proportional to levels of alpha-synuclein overexpression. Similarly, overexpression of alpha synuclein in in vitro and in vivo models has been shown to be toxic. However, little is known about the effects of reducing alpha-synuclein expression in human neurons. To investigate this, we have developed a system in which levels of alpha synuclein can be acutely suppressed by using RNA interference (RNAi) in a physiologically relevant human dopaminergic cellular model. By using small interfering RNA (siRNA) molecules targeted to endogenous alpha-synuclein, we achieved 80% protein knockdown. We show that alpha-synuclein knockdown has no effect on cellular survival either under normal growth conditions over 5 days or in the presence of the mitochondrial inhibitor rotenone. Knockdown does, however, confer resistance to the dopamine transporter (DAT)-dependent neurotoxin N-methyl 4-phenylpyridinium (MPP(+)). We then demonstrate for the first time that alpha synuclein suppression decreases dopamine transport in human cells, reducing the maximal uptake velocity (V(max)) of dopamine and the surface density of its transporter by up to 50%. These results show that RNAi-mediated alpha-synuclein knockdown alters cellular dopamine homeostasis in human cells and may suggest a mechanism for the increased survival in the presence of MPP(+), a toxin used extensively to model Parkinson's disease. PMID- 17131422 TI - Treatment of thyroid cancer based on risk groups. AB - The understanding of the prognostic factors and risk groups is crucial in the management of well differentiated thyroid cancer. The important prognostic factors are age, grade of tumor, extrathyroidal extension, size of tumor, and distant metastases. Based on these prognostic factors, patients can be divided into low-, intermediate-, and high-risk groups. The treatment decisions should be based on risk group analysis. Extent of thyroidectomy and adjuvant therapy can be based on the risk group analysis. The histopathologic studies are very important to distinguish well differentiated from the tall cell, insular, and poorly differentiated thyroid cancer. PMID- 17131423 TI - Diverging mechanisms for TNF-alpha receptors in normal mouse brains and in functional recovery after injury: From gene to behavior. AB - Cytokines, such as tumour necrosis factor (TNF)-alpha and lymphotoxin-alpha, have been described widely to play important roles in the brain in physiologic conditions and after traumatic injury. However, the exact mechanisms involved in their function have not been fully elucidated. We give some insight on their role by using animals lacking either Type 1 receptor (TNFR1KO) or Type 2 (TNFR2KO) and their controls (C57Bl/6). Both TNFR1KO and to a greater extent TNFR2KO mice showed increased exploration/activity neurobehavioral traits in the hole board test, such as rearings, head dippings, and ambulations, compared with wild-type mice, suggesting an inhibitory role of TNFR1/TNFR2 signaling. In contrast, no significant differences were observed in the elevated plus maze test, ruling out a major role of these receptors in the control of anxiety. We next evaluated the response to a freeze injury to the somatosensorial cortex. The effect of the cryolesion on motor function was evaluated with the horizontal ladder beam test, and the results showed that both TNFR1KO and TNFR2KO mice made fewer errors, suggesting a detrimental role for TNFR1/TNFR2 signaling for coping with brain damage. Expression of approximately 22600 genes was analyzed using an Affymetrix chip (MOE430A) at 0 (unlesioned), 1, or 4 days post-lesion in the three strains. The results show a unique and major role of both TNF receptors on the pattern of gene expression elicited by the injury but also in normal conditions, and suggest that blocking of TNFR1/TNFR2 receptors may be beneficial after a traumatic brain injury. PMID- 17131424 TI - Neuroprotective effect of 17beta-estradiol against N-methyl-D-aspartate-induced retinal neurotoxicity via p-ERK induction. AB - We investigated whether the neuroprotective effect of estrogen is mediated by the estrogen receptor (ER) and whether extracellular signal-regulated kinase (ERK) is involved in the protective effect of estrogen against N-methyl-D-aspartate (NMDA) induced retinal neurotoxicity. Retrograde labeling of retinal ganglion cells (RGCs) showed that pretreatment with 17beta-estradiol (E2) using a silastic implant significantly attenuated the loss of RGCs induced by intravitreal injection of NMDA. Simultaneous administration of U0126, an ERK inhibitor, with NMDA completely abolished the protective effect of E2. Moreover, ICI182,780, an ER antagonist, also significantly diminished the protective effect of E2. Pretreatment with E2 significantly reduced the number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells in the retinal ganglion cell layer (RGCL) and the inner nuclear layer (INL) 12 hr after NMDA injection. Moreover, ICI182,780 inhibited the ameliorative effect of E2 on TUNEL-positive cells in a dose-dependent manner. Immunostaining of anti-ERalpha monoclonal antibody was observed mainly in the RGCL and the INL. Western blot analysis showed a significant increase in the level of phosphorylated ERK (p-ERK) 6 hr after NMDA injection. However, NMDA did not increase the level of p-ERK protein 7 days after injection. Pretreatment of E2 induced further increases of p-ERK expression 6 hr and 7 days after NMDA injection, and U0126 and ICI182,780 significantly inhibited E2-induced p-ERK expression after 6 hr. These results suggest that E2 has an ER-mediated neuroprotective effect against NMDA-induced retinal neurotoxicity and that this effect may be associated with induction of p-ERK in the retina. PMID- 17131425 TI - Upregulation of intracellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 after unilateral nerve injury in the peripheral taste system. AB - In the peripheral taste system, activated macrophages are recruited to both sides of the tongue after unilateral sectioning of the chorda tympani nerve (CT). Neural degeneration elicits macrophage entry in other systems by upregulating vascular adhesion molecules. We hypothesized that CT sectioning leads to a bilateral increase in intracellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 expression on lingual vessels. To test this hypothesis, rats were euthanized at time points from 6 hr to 7 days post sectioning. Frozen sections of tongue were processed for immunohistochemical staining for ICAM-1 and VCAM-1. Tongue homogenates from additional rats were analyzed with ELISA. ICAM-1 expression increases first on the denervated side of the tongue at 24 hr post-section and then on the uninjured side at 48 hr post section. ICAM-1 remains elevated through Day 7 post-sectioning on both sides of the tongue. Dietary sodium restriction, which prevents the macrophage response to nerve sectioning, had no effect on ICAM-1 levels. VCAM-1+ vessels are increased on the denervated side of the tongue at 24-48 hr post-section in control-fed rats. However, dietary sodium restriction prevents the increase. These results indicate that vascular adhesion molecules are differentially regulated by CT sectioning. We suggest that macrophage entry, migration, and modulation of taste function are downstream of dynamic expression of adhesion molecules. PMID- 17131426 TI - Osmotic regulation of neuronal nitric oxide synthase expression in the rat amygdala: functional role for nitric oxide in adaptive responses? AB - Water-deprivation-induced osmotic stress leads to activation of a number of adaptive responses. Nitric oxide (NO) has been implicated in the modulation of these responses, as the amygdala has been implicated in ingestive behavior and modulation of autonomic homeostatic functions. Here we investigated the effects of water deprivation on neuronal nitric oxide synthase (nNOS) expression within the rat amygdala; a brain area involved in modulating ingestive behavior and autonomic function. Water deprivation resulted in significant increases in nNOS immunoreactivity (-ir) within different regions of the amygdala compared with euhydrated rats. Maximal increases were observed in the anteroventral (118 +/- 9 vs. 47 +/- 3 neurons), anteriodorsal (133 +/- 9 vs. 77 +/- 3), and posterioventral (175 +/- 5 vs. 71 +/- 5) parts of the medial amygdala. The basomedial nucleus (65 +/- 4 vs. 39 +/- 3) and posterior basolateral nucleus (19 +/- 2 vs. 5 +/- 1) of the amygdala and the capsular (21 +/- 2 vs. 6 +/- 1) and medial (44 +/- 6 vs. 22 +/- 3) parts of the central nucleus of the amygdala also showed increased nNOS-ir in dehydrated rats. Water deprivation had no effect on nNOS-ir in areas such as the cortical, anterior basolateral, and intercalated nuclei of the amygdala. Microinjection of an NO donor, DEA-NONOate, into the central amygdala resulted in a pressor and tachycardic response that was attenuated by a soluble guanylate cyclase inhibitor. These observations suggest that activation of the nitrergic system is prevalent throughout the amygdala following water deprivation and suggest that the up-reguation of nNOS could play a significant role in the integrative response to osmotic stress. PMID- 17131427 TI - Charged amino acid motifs flanking each extreme of the alphaM4 transmembrane domain are involved in assembly and cell-surface targeting of the muscle nicotinic acetylcholine receptor. AB - The alphaM4 transmembrane domain of the nicotinic acetylcholine receptor (AChR) is flanked by two basic amino acids (His(408) and Arg(429)) located at its cytoplasmic- and extracellular-facing extremes, respectively, at the level of the phospholipid polar head regions of the postsynaptic membrane. A series of single and double alphaM4 mutants (His(408)Ala, Arg(429)Ala, Arg(429)Glu, His(408)Ala/Arg(429)Ala, and His(408)Ala/Arg(429)Glu) of the adult muscle-type AChR were produced and coexpressed with wild-type beta, delta, and epsilon subunits as stable clones in a mammalian heterologous expression system (CHO-K1 cells). The mutants were studied by alpha-bungarotoxin ([(125)I]alpha-BTX) binding, fluorescence microscopy, and equilibrium sucrose gradient centrifugation. Cell-surface [(125)I]alpha-BTX binding diminished approximately 40% in His(408)Ala and as much as 95% in the Arg(429)Ala mutant. Reversing the amino acid charge (e.g., Arg(429)Glu) abolished cell-surface expression of AChR. Fluorescence microscopy disclosed that AChR was retained at the endoplasmic reticulum, with an enhanced occurrence of unassembled AChR species in the mutant clones. Centrifugation analysis confirmed the lack of fully assembled AChR pentamers in all mutants with the exception of His(408)Ala. We conclude that His(408) and Arg(429) in alphaM4 are involved in assembly and cell-surface targeting of muscle AChR. Arg(429) plays a more decisive role in these two processes, suggesting an asymmetric weight of the charged motifs at each extreme of the alpha subunit M4 transmembrane segment. (c) 2006 Wiley-Liss, Inc. PMID- 17131428 TI - Purification and spectroscopic characterization of the recombinant BG21 isoform of murine golli myelin basic protein. AB - A recombinant form of the murine Golli-myelin basic protein (MBP) isoform BG21 (rmBG21) has been expressed in E. coli, and isolated to 96% purity via metal chelation chromatography. Characteristic yields were 6-8 mg protein per liter of culture in either minimal M9 or standard Luria-Bertani media. Circular dichroism spectroscopy showed that rmBG21 had a large proportion of random coil in aqueous solution, but gained alpha-helix in the presence of monosialoganglioside G(M1) and PI(4)P, as well as in the membrane-mimetic solvent trifluoroethanol. Bioinformatics analyses of the amino acid sequence of rmBG21 predicted an N terminal calmodulin (CaM)-binding site. It was determined by fluorescence spectroscopy and dynamic light scattering that rmBG21 and CaM interacted weakly in a 1:1 ratio in a Ca(2+)-dependent manner. Solution NMR spectra of uniformly [(13)C(15)N]-labeled protein in aqueous buffer were consistent with it being an extended protein; spectral quality was independent of temperature. Thus, like "classic" MBP and the Golli-MBP isoform J37, rmBG21 is intrinsically disordered, implying multi functionality, and that its conformation depends on its environment and bound ligands. PMID- 17131429 TI - Selective use of radioactive iodine in the postoperative management of patients with papillary and follicular thyroid carcinoma. AB - Radioiodine remnant ablation (RRA) was developed in the 1960s to "complete a thyroidectomy" in the initial management of papillary and follicular thyroid cancer. By the 1990s, it was claimed that RRA diminished recurrence rates in follicular cell-derived cancer (FCDC) patients and decreased the cause-specific mortality (CSM) in patients more than 40 years old at initial surgery. The international trend for the past decade has been towards routine RRA in most FCDC patients. Clinical guidelines have been produced by many societies, promoting such an aggressive stance. Since 1997, many papers have reported improved outcome in FCDC, when patients were subjected to RRA after bilateral lobar resection. However, during the same time-period, it has been recognized that most FCDC patients are truly at "low-risk" of developing life-threatening recurrences. Accordingly, it has been suggested that rational therapy selection should lead to restricting aggressive therapy to those "high-risk" FCDC patients, more predisposed to CSM. To date, no prospective controlled trials exist. Presently available outcome data is based on single institutional or multicenter retrospective studies. This article summarizes the available relevant reported data, and concludes that a selective use of RRA in the postoperative management of FCDC patients is rational, and should actually be encouraged. PMID- 17131430 TI - Protein dynamics tightly connected to the dynamics of surrounding and internal water molecules. AB - Proteins are key components of biological cells. For example, enzymes catalyze biochemical reactions, membrane transporters are responsible for uptake and release of critical and superfluous components from the cell environment, and structural proteins are responsible for the stability of the cell wall and cytoskeleton. Many of the diverse protein functions involve dynamic transitions ranging from small local atomic displacements up to large allosteric conformational changes. In any conformation, proteins are in contact with the universal solvent medium of cells, water. Water not only surrounds proteins but is often an integral part of proteins and also is involved in key mechanistic steps. This Minireview discusses recent experimental and theoretical results on the role of water for protein dynamics and function. PMID- 17131431 TI - The mechanism of the molecular reorientation in water. PMID- 17131432 TI - Vertically aligned antimony nanowires as solid-state pH sensors. PMID- 17131433 TI - Size-dependent structural evolution and chemical reactivity of gold clusters. AB - Ground-state structures and other experimentally relevant isomers of Au(15) (-) to Au(24) (-) clusters are determined through joint first-principles density functional theory and photoelectron spectroscopy measurements. Subsequent calculations of molecular O(2) adsorption to the optimal cluster structures reveal a size-dependent reactivity pattern that agrees well with earlier experiments. A detailed analysis of the underlying electronic structure shows that the chemical reactivity of the gold cluster anions can be elucidated in terms of a partial-jellium picture, where delocalized electrons occupying electronic shells move over the ionic skeleton, whose geometric structure is strongly influenced by the directional bonding associated with the highly localized "d-band" electrons. PMID- 17131434 TI - A comparison of methods to compute the potential of mean force. AB - Most processes occurring in a system are determined by the relative free energy between two or more states because the free energy is a measure of the probability of finding the system in a given state. When the two states of interest are connected by a pathway, usually called reaction coordinate, along which the free-energy profile is determined, this profile or potential of mean force (PMF) will also yield the relative free energy of the two states. Twelve different methods to compute a PMF are reviewed and compared, with regard to their precision, for a system consisting of a pair of methane molecules in aqueous solution. We analyze all combinations of the type of sampling (unbiased, umbrella-biased or constraint-biased), how to compute free energies (from density of states or force averaging) and the type of coordinate system (internal or Cartesian) used for the PMF degree of freedom. The method of choice is constraint bias simulation combined with force averaging for either an internal or a Cartesian PMF degree of freedom. PMID- 17131435 TI - On the polymorphism of aspirin. PMID- 17131436 TI - Substrate modulation of the activity of an artificial nanoesterase made of peptide-functionalized gold nanoparticles. PMID- 17131437 TI - Threading cesium ions: metal, host, and ligand control in supramolecular coordination chemistry. PMID- 17131438 TI - Acid initiation of ammonia-borane dehydrogenation for hydrogen storage. PMID- 17131439 TI - [Mn(tmc)(O2)]+: a side-on peroxido manganese(III) complex bearing a non-heme ligand. PMID- 17131440 TI - Revisiting the electronic ground state of copper corroles. PMID- 17131441 TI - Formation and structure of a sterically protected molybdenum hydride complex with a 15-electron configuration: [(1,2,4-C5H2tBu3)Mo(PMe3)2H]+. PMID- 17131442 TI - An aqueous rechargeable lithium battery with good cycling performance. PMID- 17131443 TI - Functional architecture of HCV IRES domain II stabilized by divalent metal ions in the crystal and in solution. PMID- 17131445 TI - Shaping mesoporous silica nanoparticles by disassembly of hierarchically porous structures. PMID- 17131444 TI - A highly regio- and stereoselective vinylogous Horner-Wadsworth-Emmons route to densely substituted 1,3-butadienes. PMID- 17131446 TI - Rate and mechanism of the reaction of alkenes with aryl palladium complexes ligated by a bidentate P,P ligand in Heck reactions. AB - The regioselectivity of the Heck reaction is supposed to be highly affected by the electronic properties of the alkene and the ionic or neutral character of the aryl palladium(II) complexes involved in the reaction with alkenes. In Heck reactions performed in dmf, [Pd(dppp){dppp(O)}Ph](+) (dppp=1,2 bis(diphenylphosphino)propane) is generated in the oxidative addition of PhI with [Pd(0)(dppp)(OAc)](-) formed in situ from Pd(OAc)(2) associated to two equivalents of dppp. [Pd(dppp){dppp(O)}Ph](+) is not very reactive with alkenes (styrene or methyl acrylate); however, it reacts with iodide ions (released in the catalytic reactions) to give [Pd(dppp)IPh] and with acetate ions (used as base) to give [Pd(dppp)(OAc)Ph]. [Pd(dppp)(OAc)Ph] reacts with styrene and methyl acrylate exclusively by an ionic mechanism, that is, via the cationic complex [Pd(dppp)(dmf)Ph](+) formed by dissociation of the acetate ion. The reaction of [Pd(dppp)IPh] is more complex and substrate dependent. It reacts with styrene exclusively by the ionic mechanism via [Pd(dppp)(dmf)Ph](+). [Pd(dppp)IPh] (neutral mechanism) and [Pd(dppp)(dmf)Ph](+) (ionic mechanism) react in parallel with methyl acrylate. [Pd(dppp)(dmf)Ph](+) is more reactive than [Pd(dppp)IPh] but is always generated at lower concentration. PMID- 17131447 TI - An easy-to-use, regioselective, and robust bis(amidate) titanium hydroamination precatalyst: mechanistic and synthetic investigations toward the preparation of tetrahydroisoquinolines and benzoquinolizine alkaloids. AB - Amidate-supported titanium amido complexes are efficient and regioselective precatalysts for intermolecular hydroamination of terminal alkynes with primary amines. The synthesis and characterization of the first bis(amidate)-supported titanium-imido complex is reported. Its role as the active catalytic species is suggested in the course of product distribution studies using deuterated substrates. The bis(amidate)-supported precatalysts exhibit good functional-group tolerance, even performing hydroaminations in the presence of ester and amide groups. This functional-group tolerance was exploited in the synthesis of a variety of 1-substituted tetrahydroisoquinoline alkaloids and a one-pot hydroaminative procedure for the high yielding preparation of the benzo[a]quinolizine skeleton. PMID- 17131448 TI - DNA-based phosphane ligands. AB - In order to expand the repertoire of DNA sequences specifically interacting with transition metals, we report here the first examples of DNA sequences carrying mono- and bidentate phosphane ligands as well as P,N-ligands. Aminoalkyl-modified oligonucleotides have been reacted at predetermined internal sites with carboxylate derivatives of pyrphos, BINAP and phosphinooxazoline (PHOX) 2 b-d. Carbodiimide coupling in the presence of N-hydroxysuccinimide provided the DNA ligand conjugates in 38-78 % yield. Phosphane-containing oligonucleotides and their phosphane sulfide analogues were characterized by mass spectrometry (MALDI TOF and FT-ICR-ESI) and their stability after purification and isolation was systematically investigated. While DNA-appended pyrphos ligand was quickly oxidized, BINAP and PHOX conjugates showed high stabilities, making them useful precursors for incorporation of transition metals into DNA. PMID- 17131449 TI - Porous cobalt(II)-organic frameworks with corrugated walls: Structurally robust gas-sorption materials. PMID- 17131450 TI - Molecular dynamics simulation of benzene diffusion in MOF-5: importance of lattice dynamics. PMID- 17131455 TI - Toxicity of the AGEs generated from the Maillard reaction: on the relationship of food-AGEs and biological-AGEs. AB - Advanced glycation end products (AGEs) are generated in the late stages of Maillard reaction in foods and biological systems. These products are mostly formed by the reactions of reducing sugar or degradation products of carbohydrates, lipids, and ascorbic acid. AGEs exist in high concentration in foods, but in relatively low concentrations in most of the biological systems. Recently, some AGEs have been reported to be toxic, and were proposed to be causative factors for various kinds of diseases, especially diabetes and kidney disorder, through the association with receptor of AGE (RAGE). It has also been reported that food-derived AGEs (food-AGEs) may not be a causative factor for pro oxidation. However, the relationship of food-AGEs and biological-derived AGEs (biological-AGEs) is not clear. In this review, the following issues are discussed: the formation of AGEs in foods and biological systems; identification of the main AGEs in foods and biological systems; absorption of food-AGEs; the effects of AGEs in vivo; relationship between food-AGEs and biological-AGEs; possible defense mechanism against AGEs in vivo and finally, the problems to be solved concerning the toxicity of AGEs. PMID- 17131456 TI - Heterocyclic amines: chemistry and health. AB - Heterocyclic amines (HAs) occur at the ppb range in foods. Most of them demonstrate potent mutagenicity in bacteria mutagenicity test, and some of them have been classified by the International Agency for Research on Cancer as probable/possible human carcinogens. Their capability of formation even during ordinary cooking practices implies frequent exposure by the general public. Over the past 30 years, numerous studies have been stimulated aiming to alleviate human health risk associated with HAs. These studies contribute to the understanding of their formation, characterization, and quantification in foods; their mutagenesis/carcinogenesis, mechanisms of antimutagenesis by chemical or phytogenic modulators; and strategies to inhibit their formation. The chemistry of HAs, their implications in human health, factors influencing their formation, and feasible ways of suppression will be briefly reviewed. Their occurrence in trace amounts in foods necessitates continuous development and amelioration of analytical techniques. Various inhibitory strategies, ranging from modifying cooking conditions to incorporation of different modulators, have been developed. This will remain one of the foremost areas of research in the field of food chemistry and safety. PMID- 17131457 TI - Chemistry and some biological effects of model melanoidins and pigments as Maillard intermediates. AB - Various pigments were formed in the D-xylose-glycine reaction system. Blue pigments (Blue-M1 and Blue-M2) and red pigments (Red-M1 and Red-M2) were generated in the Maillard reaction. Blue-M2 is presented to have been generated by the reaction between Blue-M1, which involved two pyrrolopyrrole structures as the major blue pigment, and di-D-xyluloseglycine. We identified red pigments as the isomers of addition compounds of D-xyluloseglycine to condensated compound between pyrroropyrrole-2-carbaldehyde and pyrrole-2-carbaldehyde compounds. These pigments have polymerizing activities, suggesting that they are important Maillard reaction intermediates through the formation of melanoidins. Blue-M1 as well as melanoidins effectively suppressed the peroxidation of linoleic acid. The scavenging activity toward Blue-M1 on hydroxyl and DPPH radicals was also as strong as that of melanoidins. Furthermore, Blue-M1 prevents the oxidative cell injury. Therefore, Blue-M1 will be an antioxidant which protects against the oxidative stress in biological systems. Melanoidins induced IFN-gamma mRNA and IL 12 mRNA expressions in spleen cells exposed to allergen and in macrophage-like J774.1 cells, respectively. These findings suggest that melanoidins have suppressive effect on allergic reaction as a novel physiological effect. PMID- 17131458 TI - Intervention with polyphenol-rich fruit juices results in an elevation of glutathione S-transferase P1 (hGSTP1) protein expression in human leucocytes of healthy volunteers. AB - Polyphenols are probably antigenotoxic on account of their antioxidant activities and might alter phase I and II enzymes in a way that results in chemoprotection. We investigated the hypothesis that polyphenols enhance expression of glutathione S-transferases (GSTs), which increases carcinogen detoxification and thereby provides protection against oxidative stress. HGSTP1 protein expression and GST polymorphisms were determined in leucocytes obtained during an intervention study with healthy subjects consuming two fruit juices in an 8 wk trial (polyphenol free run in phase, juice intervention phase, washout phase, second juice intervention phase, each treatment regime lasted for 2 wk). The study had originally shown that juice intervention significantly reduced oxidative DNA damage in leucocytes at week 8 (Bub, A., Watzl, B., Blockhaus, M., Briviba, K. et al., J. Nutr. Biochem. 2003, 14, 90-98). We reanalysed the levels of DNA damage based on GST genotypes. We also treated leucocytes in vitro with mixtures of polyphenols and determined cytotoxicity and expression of 96 genes related to drug metabolism. Key results with leucocytes of the intervention study were that the initial content of hGSTP1 protein was first suppressed at weeks 4 and 6. At week 8, however, hGSTP1 protein expression was significantly increased. HGSTP1 protein levels and DNA damage were inversely correlated (p = 0.005), but there was no difference for cells obtained from subjects with hGSTM1*1 and hGSTM1*0 genotypes, nor was there any difference between cells from subjects consuming the two different juices. The treatment of leucocytes with polyphenol mixtures in vitro did not result in modulated GST gene expression or total GST activity, but in an up-regulation of other biotransformation enzymes (e. g., members of the cytochrome P450 and the sulphotransferase family). In conclusion, in vitro treatment of leucocytes led to a modulated mRNA expression of selected genes, not directly related to oxidative defence systems. In vivo, however, we observed a delayed enhancement of hGSTP1, which could be associated with an initial repression of oxidative DNA damage in leucocytes from human subjects, consuming juices with high levels of polyphenols. PMID- 17131459 TI - The potential involvement of glutamate ingestion in chronic neurodegenerative diseases. Opinion of the Senate Commission on Food Safety (SKLM) of the German Research Foundation (DFG)-(shortened version). PMID- 17131461 TI - Benzo[d]isothiazol-3-yl-benzamidines: a class of protective agents on culture of human cartilage and chondrocytes stimulated by IL-1beta. AB - New derivatives of N-benzo[d]isothiazol-3-yl-benzamidine 6 a were synthesized as nonacidic anti-inflammatory/antidegenerative agents. We investigated the influence of the amidines 6 a-j on the production of NO, PGE(2), MMP-3, COX-2, ROS, and GAGs, key molecules involved in cartilage destruction in osteoarthritic diseases. The antidegenerative properties of the novel designed derivatives 6 b-j were improved with respect to N-benzo[d]isothiazol-3-yl-benzamidine 6 a. All of the compounds 6 a-j promoted the reduction of most of the IL-1beta-induced harmful effects. Derivatives 6 d, 6 h, and 6 j were the most potent of all the tested compounds, particularly in the human chondrocyte culture model. PMID- 17131462 TI - Design, syntheses, biological evaluation, and docking studies of 2-substituted 5 methylsulfonyl-1-phenyl-1H-indoles: potent and selective in vitro cyclooxygenase 2 inhibitors. AB - Four series of 5-methylsulfonyl-1-phenyl-1H-indole-2-carboxylic acid alkyl esters (family A), -2-carbonitriles (family B), -2-carboxamides (family C), and 2 benzoyl-5-methylsulfonyl-1-phenyl-1H-indoles (family D) were prepared and evaluated for their ability to inhibit purified cyclooxygenase-2 (COX-2) and cyclooxygenase-1 (COX-1). Family D compounds have the best COX-1/COX-2 inhibition ratios and potencies. According to docking studies, these molecules appear to bind the COX-2 binding site differently than indomethacin, with the insertion of the substituent at the 2-position in the hydrophobic pocket of the enzyme and the 1-position phenyl ring in the trifluoromethyl zone. Among the group of compounds evaluated, 2-(4-chlorobenzoyl)-1-(4-chlorophenyl)-5-methylsulfonyl-1H-indole and 2-(4-chlorophenyl)-5-methylsulfonyl-1-(4-trifluoromethylphenyl)-1H-indole emerged as the most potent (respective IC(50) values: 46 and 43 nM), and selective (respective selectivity indexes: >2163 and >2331) COX-2 inhibitors. PMID- 17131463 TI - Macrocyclic aminopyrimidines as multitarget CDK and VEGF-R inhibitors with potent antiproliferative activities. AB - X-ray structures from CDK2-aminopyrimidine inhibitor complexes led to the idea to stabilize the active conformation of aminopyrimidine inhibitors by incorporating the recognition site into a macrocyclic framework. A modular synthesis approach that relies on a new late-stage macrocyclization protocol that enables fast and efficient synthesis of macrocyclic aminopyrimidines was developed. A set of structurally diverse derivatives was prepared. Macrocyclic aminopyrimidines were shown to be multitarget inhibitors of CDK1/2 and VEGF-RTKs. In addition, potent antiproliferative activities toward various human tumor cells and a human tumor xenograft model were demonstrated. PMID- 17131464 TI - Identification of endogenous resolvin E1 and other lipid mediators derived from eicosapentaenoic acid via electrospray low-energy tandem mass spectrometry: spectra and fragmentation mechanisms. AB - Resolvin E1 (RvE1, 5S,12R,18R-trihydroxy-6Z,8E,10E,14Z,16E-eicosapentaenoic acid) is a novel anti-inflammatory lipid mediator recently found in humans, mice, and fish in vivo. To identify endogenous RvE1 and other eicosapentaenoic acid (EPA) derived lipid mediators using electrospray low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), the MS/MS product ion spectra of these compounds were correlated with their structures, and the MS/MS fragmentation mechanisms were studied. Deuterium labeling confirmed the proposed correlations and the fragmentation mechanisms. beta-cleavage was observed for RvE1, and beta and gamma cleavages were seen for leukotriene B5; however, alpha cleavage was more common. The positions and numbers of hydroxyls and double bonds of these lipid mediators can be deduced from the MS/MS spectra. The MS/MS fragmentation generating chain-cut ions involved beta-ene, gamma-ene, or alpha-H beta-ene rearrangement, depending on the specific structure. The m/z value of a detected chain-cut ion from RvE1 or from an EPA-derived product is equal to the corresponding hypothetical homolytic segment (cc, cm, mc, or mm) with the addition or extraction of up to two hydrogen atoms (H) from hydroxyls or an alpha carbon; namely, the m/z value of an alpha-cleavage-generated ion is equal to [cc+H], [cm-2H], [mc-H], or [mm]. Wideband activation increased the signal intensities of chain-cut ions, and therefore was better for trace analysis of RvE1 in biological samples. RvE1, LTB5, PGE3, and other EPA-derived lipid mediators were found in trout brain or head-kidney via this approach on the basis of MS/MS spectra and fragmentation mechanisms. Negative ion electrospray low collision-energy MS/MS spectra provide adequate data to elucidate and identify the structures of RvE1 and other EPA-derived lipid mediators at levels below a few picomoles in trout samples. PMID- 17131465 TI - A novel method for the study of the biophysical interface in soils using nano scale secondary ion mass spectrometry. AB - The spatial location of microorganisms and their activity within the soil matrix have major impacts on biological processes such as nutrient cycling. However, characterizing the biophysical interface in soils is hampered by a lack of techniques at relevant scales. A novel method for studying the distribution of microorganisms that have incorporated isotopically labelled substrate ('active' microorganisms) in relation to the soil microbial habitat is provided by nano scale secondary ion mass spectrometry (NanoSIMS). Pseudomonas fluorescens are ubiquitous in soil and were therefore used as a model for 'active' microorganisms in soil. Batch cultures (NCTC 10038) were grown in a minimal salt medium containing 15N-ammonium sulphate (15/14N ratio of 1.174), added to quartz-based white sand or soil (coarse textured sand), embedded in Araldite 502 resin and sectioned for NanoSIMS analysis. The 15N-enriched P. fluorescens could be identified within the soil structure, demonstrating that the NanoSIMS technique enables the study of spatial location of microbial activity in relation to the heterogeneous soil matrix. This technique is complementary to the existing techniques of digital imaging analysis of soil thin sections and scanning electron microscopy. Together with advanced computer-aided tomography of soils and mathematical modelling of soil heterogeneity, NanoSIMS may be a powerful tool for studying physical and biological interactions, thereby furthering our understanding of the biophysical interface in soils. PMID- 17131466 TI - Biofeedback therapy for dyssynergic defecation. AB - Dyssynergic defecation is one of the most common forms of functional constipation both in children and adults; it is defined by incomplete evacuation of fecal material from the rectum due to paradoxical contraction or failure to relax pelvic floor muscles when straining to defecate. This is believed to be a behavioral disorder because there are no associated morphological or neurological abnormalities, and consequently biofeedback training has been recommended for treatment. Biofeedback involves the use of pressure measurements or averaged electromyographic activity within the anal canal to teach patients how to relax pelvic floor muscles when straining to defecate. This is often combined with teaching the patient more appropriate techniques for straining (increasing intra abdominal pressure) and having the patient practice defecating a water filled balloon. In adults, randomized controlled trials show that this form of biofeedback is more effective than laxatives, general muscle relaxation exercises (described as sham biofeedback), and drugs to relax skeletal muscles. Moreover, its effectiveness is specific to patients who have dyssynergic defecation and not slow transit constipation. However, in children, no clear superiority for biofeedback compared to laxatives has been demonstrated. Based on three randomized controlled studies in the last two years, biofeedback appears to be the preferred treatment for dyssynergic defecation in adults. PMID- 17131467 TI - Insulin resistance and hepatitis C. AB - Insulin resistance is the major feature of the metabolic syndrome and depends on insulin secretion and insulin sensitivity. In chronic hepatitis C, insulin resistance and type 2 diabetes mellitus are more often seen than in healthy controls or chronic hepatitis B patients. Hepatitis C virus (HCV) infection promotes insulin resistance, mainly by increased TNF production together with enhancement of suppressor of cytokine (SOC-3); both events block PI3K and Akt phosphorylation. Two types of insulin resistance could be found in chronic hepatitis C patients: "viral" and "metabolic" insulin resistance. Insulin resistance in chronic hepatitis C is relevant because it promotes steatosis and fibrosis. The mechanisms by which insulin resistance promotes fibrosis progression include: (1) steatosis, (2) hyperleptinemia, (3) increased TNF production, (4) impaired expression of PPARgamma receptors. Lastly, insulin resistance has been found as a common denominator in patients difficult-to-treat like cirrhotics, overweight, HIV coinfected and Afro-American. Insulin resistance together with fibrosis and genotype has been found to be independently associated with impaired response rate to peginterferon plus ribavirin. Indeed, in genotype 1, the sustained response rate was twice (60%) in patients with HOMA < or = 2 than patients with HOMA > 2. In experiments carried out on Huh-7 cells transfected by full length HCVRNA, interferon alpha blocks HCV replication. However, when insulin (at doses of 128 microU/mL, similar that seen in the hyperinsulinemic state) was added to interferon, the ability to block HCV replication disappeared, and the PKR synthesis was abolished. In summary, hepatitis C promotes insulin resistance and insulin resistance induces interferon resistance, steatosis and fibrosis progression. PMID- 17131468 TI - The "return" of hepatitis B. AB - There has been a significant advance in the treatment of chronic Hepatitis B virus (HBV) infection and the following drugs were approved for therapy: Conventional interferon (IFN), pegylated interferon alfa-2a (PEG IFN alpha2a), lamivudine, adefovir and entecavir. Compared to nucleoside analogues IFN induces higher rates of sustained remission and HBsAg loss. Conventional IFN in lower doses (1, 5-3 MIU) tiw for 4-6 mo has similar efficacy in comparison to "standard IFN therapy". Longer IFN treatment is a significant factor for long-term remission in HBeAg-negative CHB, but the higher actual IFN dose is not such a factor. PEG IFN is superior to conventional IFN. There is no significant difference between PEG IFN alpha2a at doses 90 mcg/wk and 180 mcg/wk in HBeAg positive patients. These results provide a rational for further clinical trials with lower doses PEG IFN alpha2a given in prolonged course as maintenance or intermittent treatment. Serious new problems arose after the introduction of nucleoside/nucleotide analogues in clinical practice. The most important ones are drug-resistance and the high rates of relapse after treatment discontinuation. Therapy should only be recommended if the expected benefit exceeds significantly the abstain from treatment. The choice of therapy should take into account the patient's age, co-morbidity, severity of liver disease and the risk of drug resistance. New antivirals significantly suppress HBV-replication, but have no effect on cccDNA in hepatocytes, and after the treatment discontinuation viral relapses occurs. At the present level of knowledge it is impossible "to eradicate the virus" The realistic treatment goal is to achieve durable response by clearance of HBeAg, sustained decrease of serum HBV DNA levels, normalization of ALT, improvement of liver histology and stopping of liver fibrogenesis. The competition between IFN based therapy and nucleoside or nucleotide analogues still remains. IFN can cure the liver disease while nucleotide analogues only suppress the viral replication during therapy and can reduce the liver fibrosis. Treatment should be prolonged for 24-mo or longer by using maintenance or intermittent treatment course with the lowest effective IFN and PEG IFN doses. Nucleoside/nucleotide analogues are a promising treatment option, but additional data for treatment duration and long-term post-treatment outcome are necessary. PMID- 17131469 TI - Pathophysiology of pulmonary complications of acute pancreatitis. AB - Acute pancreatitis in its severe form is complicated by multiple organ system dysfunction, most importantly by pulmonary complications which include hypoxia, acute respiratory distress syndrome, atelectasis, and pleural effusion. The pathogenesis of some of the above complications is attributed to the production of noxious cytokines. Clinically significant is the early onset of pleural effusion, which heralds a poor outcome of acute pancreatitis. The role of circulating trypsin, phospholipase A2, platelet activating factor, release of free fatty acids, chemoattractants such as tumor necrosis factor (TNF)-alpha, interleukin (IL)-1, IL-6, IL-8, fMet-leu-phe (a bacterial wall product), nitric oxide, substance P, and macrophage inhibitor factor is currently studied. The hope is that future management of acute pancreatitis with a better understanding of the pathogenesis of lung injury will be directed against the production of noxious cytokines. PMID- 17131470 TI - Associations between NOD2/CARD15 genotype and phenotype in Crohn's disease--Are we there yet? AB - There have been multiple NOD2/CARD15 genotype-phenotype analyses undertaken in patients with Crohn's disease since the gene's discovery in 2001. This review focuses on the major published series based upon their size and on the presence of specific clinical and genetic information provided in the published material from 2001 to 2005. Twelve studies provided raw data to carry out comparisons of disease location while ten studies included analysis of NOD2/CARD15 genotypes. NOD2/CARD15 variant frequency in ileal disease did not differ significantly among studies, whereas a comparison of disease location demonstrated highly significant differences among studies. Meta-analysis confirmed significant associations between NOD2/CARD15 variants and both ileal and ileocolonic disease locations, and with both stricturing and penetrating forms of disease behavior. This review underlines the significant phenotypic differences that exist among populations, including similar ethnic groups, and has demonstrated the need for further studies of patients with long-term "inflammatory" Crohn's disease. PMID- 17131471 TI - Identification of squamous cell carcinoma associated proteins by proteomics and loss of beta tropomyosin expression in esophageal cancer. AB - AIM: To assess the proteome of normal versus tumor tissue in squamous cell carcinoma of the esophagus (SCCE) in Iranian patients and compare our results with former reports by using proteomics. METHODS: Protein was extracted from normal and tumor tissues. Two dimensional electrophoresis was carried out and spots with differential expression were identified with mass spectrometry. RNA extraction and RT-PCR along with immunodetection were performed. RESULTS: Fourteen proteins were found whose expression levels differed in tumor compared to normal tissues. Mass spectrometric analysis resulted in the identification of beta-tropomyosin (TMbeta), myosin light chain 2 (and its isoform), myosin regulatory light chain 2, peroxyredoxin 2, annexin I and an unknown polypeptide as the down regulated polypeptides in tumor tissue. Heat shock protein 70 (HSP70), TPM4-ALK fusion oncoprotein 2, myosin light polypeptide 6, keratin I, GH16431p and calreticulin were the up-regulated polypeptides found in tumor tissue. Several of these proteins, such as TMbeta, HSP70, annexin I, calreticulin, TPM4-ALK and isoforms of myosins, have been well recognized in tumorigenesis of esophageal or other types of cancers. CONCLUSION: Our study not only supports the involvement of some of the formerly reported proteins in SCCE but also introduces additional proteins found to be lost in SCCE, including TMbeta. PMID- 17131472 TI - Hypermethylation and expression regulation of secreted frizzled-related protein genes in colorectal tumor. AB - AIM: To investigate the functions of promoter hyper-methylation of secreted frizzled-related proteins (sFRPs) genes in colorectal tumorigenesis and progression. METHODS: The promoter hypermethylation and expression of sFRP genes in 72 sporadic colorectal carcinomas, 33 adenomas, 18 aberrant crypt foci (ACF) and colorectal cancer cell lines RKO, HCT116 and SW480 were detected by methylation-specific PCR and reverse transcription PCR, respectively. RESULTS: None of the normal colorectal mucosa tissues showed methylated bands of any of four sFRP genes. sFRP1, 2, 4 and 5 were frequently methylated in colorectal carcinoma, adenoma and ACF (sFRP1 > 85%, sFRP2 > 75%, sFRP5 > 50%), and the differences between three colorectal tissues were not significant (P > 0.05). Methylation in colorectal tumors was more frequent than in normal mucosa and adjacent normal mucosa. The mRNA of sFRP1-5 genes was expressed in all normal colorectal mucosa samples. Expression of sFRP1, 2, 4 and 5 and sFRP1, 2 and 5 was downregulated in carcinoma and adenoma, respectively. The downregulation of sFRP2, 4 and 5 was more frequent in carcinoma than in adenoma. Expression of sFRP3 which promoter has no CpG island was downregulated in only a few of colorectal tumor samples (7/105). The downregulation of sFRP1, 2, 4 and 5 expression was significantly associated with promoter hypermethylation in colorectal tumor. After cells were treated by DAC/TSA combination, the silenced sFRP mRNA expression could be effectively re-expressed in colorectal cancer cell lines. CONCLUSION: Hypermethylation of sFRP genes is a common early event in the evolution of colorectal tumor, occurring frequently in ACF, which is regarded as the earliest lesion of multistage colorectal carcinogenesis. It appears to functionally silence sFRP genes expression. Methylation of sFRP1, 2 and 5 genes might serve as indicators for colorectal tumor. PMID- 17131473 TI - Genetic vaccination with Flt3-L and GM-CSF as adjuvants: Enhancement of cellular and humoral immune responses that results in protective immunity in a murine model of hepatitis C virus infection. AB - AIM: To investigate whether transfection of plasmid DNA encoding these cytokines enhances both humoral and cellular immune responses to hepatitis C virus (HCV) in a murine model. METHODS: We established a tumor model of HCV infection using syngenic mouse myeloma cells stably transfected with NS5. Co-vaccination of DNA encoding granulocyte macrophage colony-stimulating factor (GM-CSF) and Flt-3 ligand together with a plasmid encoding for the HCV NS5 protein was carried out. Mice were sacrificed 14 d after the last immunization event with collection of spleen cells and serum to determine humoral and cellular immune responses. RESULTS: Co-vaccination of DNA encoding GM-CSF and Flt-3 ligand together with a plasmid encoding for the HCV NS5 protein induced increased antibody responses and CD4+ T cell proliferation to this protein. Vaccination with DNA encoding GM-CSF and Flt-3L promoted protection against tumor formation and/or reduction in mice co-immunized with cytokine-encoding DNA constructs. This suggests this strategy is capable of generating cytotoxic T lymphocyte activity in vivo. Following inoculation with plasmid DNA encoding Flt-3L, no increase in spleen size or in dendritic cell (DC) and natural killer cell numbers was observed. This was in contrast to a dramatic increase of both cell types after administration of recombinant Flt3-L in vivo. This suggests that vaccination with plasmid DNA encoding cytokines that regulate DC generation and mobilization may not promote unwanted side effects, such as autoimmunity, splenic fibrosis or hematopoietic malignancies that may occur with administration of recombinant forms of these proteins. CONCLUSION: Our data support the view that plasmid DNA vaccination is a promising approach for HCV immunization, and may provide a general adjuvant vaccination strategy against malignancies and other pathogens. PMID- 17131475 TI - Antimicrobial activity of Sapindus mukorossi and Rheum emodi extracts against H pylori: In vitro and in vivo studies. AB - AIM: to evaluate the antibacterial activity of Sapindus mukorossi (S. mukorossi) and Rheum emodi (R. emodi). METHODS: Powders of S. mukorossi and R. emodi were extracted successively with petroleum ether, benzene, chloroform and ethanol and were concentrated in vacuum. The disk diffusion method was used for in vitro studies and in vivo studies were performed on male Wister rats. Thirty resistant clinical isolates of H pylori, as determined by their antibiotic sensitivity patterns by E-test, along with two Gram +ve (S. aureus, B. subtilis) and two Gram -ve (E. coli, P. vugaris) organisms were screened for their susceptibility patterns against these extracts. RESULTS: In our screening, all 30 resistant isolates and the other four organisms (two Gram +ve S. aureus, B. subtilis and two Gram -ve, E. coli, P. vugaris) were sensitive to the test compounds. It was found that ethanol and chloroform extracts of S. mukorossi and ethanol and benzene extracts of R. emodi inhibited H pylori at very low concentrations. In the in vitro study, the isolates showed a considerable zone of inhibition at very low concentrations (10 mug/mL) for both the extracts. In the in vivo study, the H pylori infection was cleared with minimal doses of extracts of S. mukorossi (2.5 mg/mL) and R. emodi (3.0 mg/mL) given orally for seven days. CONCLUSION: We can conclude from this study that the extracts of S. mukorossi and R. emodi inhibited the growth of pylori in vitro and, in in vivo studies, the H pylori infection cleared within seven days at very low concentrations. We also found that H pylori did not acquire resistance against these herbal extracts even after 10 consecutive passages. PMID- 17131474 TI - Elicitation of strong immune responses by a DNA vaccine expressing a secreted form of hepatitis C virus envelope protein E2 in murine and porcine animal models. AB - AIM: To characterize the immunogenicity of a hepatitis C virus (HCV) E2 DNA vaccine alone or with a protein vaccine boost in murine and porcine animal models. METHODS: A DNA vaccine expressing a secreted form of HCV E2 protein was constructed and used to vaccinate mice and piglets with or without boosting with a recombinant E2 protein vaccine formulated with CpG ODN and 10% Emulsigen. The immunogenicity of HCV E2 vaccines was analyzed by ELISA for antibody responses, MTT assay for lymphocyte proliferation, ELISPOT for the number of interferon gamma secreting cells, and cytotoxic T lymphocyte assays. RESULTS: Intradermal injection of E2 DNA vaccine induced strong Th1-like immune responses in mice. In piglets, E2 DNA vaccine elicited moderate and more balanced immune responses. A DNA vaccine prime and protein boost vaccination strategy induced significantly higher E2-specific antibody levels and shifted the immune response towards Th2 like ones in piglets. CONCLUSION: A DNA vaccine expressing a secreted form of HCV E2 protein elicited E2-specific immune responses in mice and piglets. Recombinant E2 protein vaccination following DNA immunization significantly increased the antibody response in piglets. These HCV E2 vaccines may represent promising hepatitis C vaccine candidates for further investigations. PMID- 17131476 TI - Elenoside increases intestinal motility. AB - AIM: To study the effects of elenoside, an arylnaph-thalene lignan from Justicia hyssopifolia, on gastro-intestinal motility in vivo and in vitro in rats. METHODS: Routine in vivo experimental assessments were catharsis index, water percentage of boluses, intestinal transit, and codeine antagonism. The groups included were vehicle control (propylene glycol-ethanol-plant oil-tween 80), elenoside (i.p. 25 and 50 mg/kg), cisapride (i.p. 10 mg/kg), and codeine phosphate (intragastric route, 50 mg/kg). In vitro approaches used isolated rat intestinal tissues (duodenum, jejunum, and ileum). The effects of elenoside at concentrations of 3.2 x 10(-4), 6.4 x 10(-4) and 1.2 x 10(-3) mol/L, and cisapride at 10(-6) mol/L were investigated. RESULTS: Elenoside in vivo produced an increase in the catharsis index and water percentage of boluses and in the percentage of distance traveled by a suspension of activated charcoal. Codeine phosphate antagonized the effect of 25 mg/kg of elenoside. In vitro, elenoside in duodenum, jejunum and ileum produced an initial decrease in the contraction force followed by an increase. Elenoside resulted in decreased intestinal frequency in duodenum, jejunum, and ileum. The in vitro and in vivo effects of elenoside were similar to those produced by cisapride. CONCLUSION: Elenoside is a lignan with an action similar to that of purgative and prokinetics drugs. Elenoside, could be an alternative to cisapride in treatment of gastrointestinal diseases as well as a preventive therapy for the undesirable gastrointestinal effects produced by opioids used for mild to moderate pain. PMID- 17131477 TI - Effect of Kaiyu Qingwei Jianji on the morphometry and residual strain distribution of small intestine in experimental diabetic rats. AB - AIM: To investigate the effect of a Chinese medicine, Kaiyu Qingwei Jianji (KYQWJJ) used for diabetic treatment, on the morphometry and residual strain distribution of the small intestine in streptozotocin (STZ) -induced diabetic rats. Correlation analysis was also performed between the opening angle and residual strain with the blood glucose level. METHODS: Forty-two male Wistar rats weighing 220-240 g were included in this study. Thirty-two STZ-induced diabetic rats were subdivided into four groups (n = 8 in each group), i.e. diabetic control group (DM); high dose of KYQWJJ (T1, 36 g/kg per day); low dose of KYQWJJ (T2, 17 g/kg per day) and Gliclazide (T3, 50 mg/kg per day). Another ten rats were used as non-diabetic control (CON). The medicines were poured directly into stomach lumen by gastric lavage twice daily. The rats of CON and DM groups were only poured the physiological saline. Blood glucose and plasma insulin levels were measured. Experimental period was 35 d. At the end of experiment, three 5-cm long segments were harvested from the duodenum, jejunum and ileum. Three rings of 1-2 mm in length for no-load and zero-stress state tests were cut from the middle of different segments. The morphometric data, such as the circumferential length, the wall thickness and the opening angle were measured from the digitized images of intestinal segments in the no-load state and zero-stress state. The residual strain was computed from the morphometry data. Furthermore, the linear regression analysis was performed between blood glucose level with morphometric and biomechanical data in the different intestinal segments. RESULTS: The blood glucose level of DM group was consistent 4-fold to 5-fold higher than those in CON group during the experiment (16.89+/-1.11 vs 3.44+/-0.15 mmol/L, P < 0.001). The blood glucose level in the T1 (16.89+/-1.11 vs 11.08+/-2.67 mmol/L, P < 0.01) and T3 groups (16.89+/-1.11 vs 13.54+/-1.73 mmol/L, P < 0.05), but not in T2 group (P > 0.05) was significantly lower than those in DM group. The plasma insulin levels of DM, T1, T2 and T3 groups were significantly lower than those in CON group (10.98+/-1.02, 12.52+/-1.42,13.54+/-1.56,10.96+/-0.96 vs 17.84+/-2.34 pmol/L respectively, P < 0.05), but no significantly difference among the groups with exception of CON group. The wet weight/cm and total wall thickness of duodenum, jejunum and ileum in DM group were significantly higher than those in CON group (wet weight (g/cm): duodenum 0.209+/-0.012 vs 0.166+/-0.010, jejunum 0.149+/-0.008 vs 0.121+/-0.004, ileum 0.134+/-0.013 vs 0.112+/-0.007; Wall thickness (mm): duodenum 0.849+/-0.027 vs 0.710+/-0.026, jejunum 0.7259+/-0.034 vs 0.627+/-0.025, ileum 0.532+/-0.023 vs 0.470+/-0.010, all P < 0.05), T1 and T3 treatment could partly restore change of wall thickness, but T2 could not. The opening angle and absolute value of inner and outer residual stain were significantly smaller in duodenal segment (188+/-11 degrees, -0.31+/-0.02 and 0.35+/-0.03 vs 259+/-15 degrees, -0.40+/-0.02 and 0.43+/-0.05) and larger in jejunal (215+/-20 degrees, -0.30+/-0.03 and 0.36+/-0.06 vs 172+/-19 degrees, 0.25+/-0.02 and 0.27+/-0.02) and ileal segments (183+/-20 degrees, -0.28+/-0.01 and 0.34+/-0.05 vs 153+/-14 degrees, -0.23+/-0.03 and 0.29+/-0.04) in DM group than in CON group (P < 0.01). T1 and T3 treatment could partly restore this biomechanical alteration, but strong effect was found in T1 treatment (duodenum 243+/-14 degrees, -0.36+/-0.02 and 0.42+/-0.06, jejunum 180+/-15 degrees, -0.26+/ 0.03 and 0.30+/-0.06 and ileum 163+/-17 degrees, -0.23+/-0.03 and 0.30+/-0.05, compared with DM, P < 0.05). The linear association was found between the glucose level with most morphometric and biomechanical data. CONCLUSION: KYQWJJ (high dose) treatment could partly restore the changes of blood glucose level and the remodeling of morphometry and residual strain of small intestine in diabetic rats. The linear regression analysis demonstrated that the effect of KYQWJJ on intestinal opening angle and residual strain is partially through its effect on the blood glucose level. PMID- 17131478 TI - Noninvasive evaluation of hepatic fibrosis in children with infant hepatitis syndrome. AB - AIM: To elucidate the impact of hemodynamic para-meters on ultrasonography and serum fibrosis markers for the assessment of liver fibrosis in the children with infant hepatitis syndrome (IHS). METHODS: Forty-one children with IHS and 46 healthy infants were examined by ultrasonography, and several hemodynamic indices such as peak systolic velocity (PSV) and resistant index (RI) of proper hepatic artery (PHA) were measured. Serum fibrosis markers including hyaluronic acid (HA), pre-collagen type-III (PC-III), collagen type IV (C-IV), and laminin (LN) were assayed by radioimmunoassays. In children with IHS, liver tissues were obtained either by ultrasound-guided liver biopsy (n = 35) or in the course of operation (n = 6). The stages of hepatic fibrosis were scored as mild (S1 and S2), moderate (S3), or severe (S4) according to liver histological diagnosis. Multiple groups comparative and Spearman correlative analyses were carried out. RESULTS: Histopathologically, 39 children (95.1%) were found to have hepatic fibrosis, 12 of them stage S1 or S2, 12 stage S3, and 15 stage S4. PSV, RI of the PHA, and serum HA showed a consecutive increase from mild to severe hepatic fibrosis and a close positive correlation with hepatic fibrosis in IHS group (r = 0.717, 0.745 and 0.712, respectively, P = 0.001). The Doppler waveform of HV was also positively correlated with the degree of hepatic fibrosis in IHS group (r = 0.783, P < 0.001). CONCLUSION: Combination of ultrasonic studies on the hepatic hemodynamics with the evaluation of serum HA may provide an indicator for hepatic fibrosis in patients with IHS. This may be a useful noninvasive method for the diagnosis and evaluation of the prognosis of IHS. PMID- 17131479 TI - Relationship between Crohn's disease, infection with Mycobacterium avium subspecies paratuberculosis and SLC11A1 gene polymorphisms in Sardinian patients. AB - AIM: To study the association between Crohn's disease (CD), Mycobacterium avium subspecies paratuberculosis (MAP), and genetic factors by examining the role of natural resistance-associated macrophage protein 1 (NRAMP1) gene polymorphisms (now SLC11A1) in Sardinian patients with CD and controls. METHODS: Thirty-seven CD patients and 34 controls with no inflammatory bowel disease (IBD) were recruited at the University of Sassari after giving written consent. Six SCL11A1 polymorphisms previously reported to be the most significantly associated with IBD were searched. M. paratuberculosis was identified by IS900 PCR and sequencing. Logistic regression was used to calculate odds ratios (OR) for the associations among CD, presence of MAP, and 6 loci described above. RESULTS: For the first time, a strong association was observed between polymorphisms at NRAMP1 locus 823C/T and CD. While CD was strongly associated with both NRAMP1 and MAP, NRAMP1 polymorphisms and MAP themselves were not correlated. CONCLUSION: Combined with previous work on the NOD2/CARD15 gene, it is clear that the interplay of genetic, infectious, and immunologic factors in the etiology of CD is complex. PMID- 17131480 TI - Trocar-related abdominal wall bleeding in 200 patients after laparoscopic cholecistectomy: Personal experience. AB - AIM: To determine the complications and incidence of the first and second access related vascular injuries induced by videolaparoscopic cholecistectomy. METHODS: We retrospectively reviewed vascular injuries in 200 consecutive patients who underwent videolaparoscopic cholecistectomy from 2003 to 2005. One hundred and one patients with placement of radial expanding trocars were assigned into group A and 99 patients with placement of pyramidal tipped trocars into group B. All the patients were submitted to open access according to Hasson for the first trocar. RESULTS: Bleeding did not occur at the intraoperative cannula-site in group A. However, it occurred at the intraoperative cannula-site of 7 patients (7.1%) in group B, with a statistically significant difference (P < 0.01). No mortality was registered. More vascular lesions were found in group B. CONCLUSION: The advantage of Hasson technique is that peritoneal cavity access is gained under direct vision, preventing most severe injuries. The open technique with radial expanding trocars is recommended for secure access to the abdominal cavity in videolaparoscopy. Great care should be taken to avoid major complications and understanding the abdominal wall anatomy is important for reducing bleeding during or after s placement of trocars. PMID- 17131481 TI - Protective effects of amphetamine on gastric ulcerations induced by indomethacin in rats. AB - AIM: To study the effects of amphetamine, an indirect-acting adrenomimetic compound on the indomethacin-induced gastric ulcerations in rats. METHODS: Male Wistar-Bratislava rats were randomly divided into four groups: Group 1 (control), received an ulcerogenic dose of indomethacin (50 micromol/kg) and Groups 2, 3 and 4, treated with amphetamine (10, 25 and 50 micromol/kg). The drug was administered simultaneously with indomethacin and once again 4 h later. The animals were sacrificed 8 h after indomethacin treatment. The stomachs were opened and the incidence, the number of lesions and their severity were evaluated. The results were expressed as percentage and as mean+/-standard error (mean+/-SE). RESULTS: The incidence of ulceration in the control group was 100%. Amphetamine, at doses of 10, 25 and 50 micromol/kg, lowered the incidence to 88.89%, 77.78% and 37.5% respectively. The protection ratio was positive: 24.14%, 55.17% and 80.6% respectively. The total number of ulcerations/rat was 12.44+/ 3.69 in the control group. It decreased to 7.33+/-1.89, 5.33+/-2.38 and 2.25+/ 1.97 under the effects of the above-mentioned doses of amphetamine. CONCLUSION: Amphetamine affords a significant dose-dependent protection against the indomethacin-induced gastric ulcerations in rats. It is suggested that the adrenergic system is involved in the gastric mucosa protection. PMID- 17131482 TI - Non-invasive diagnosis of gastric mucosal atrophy in an asymptomatic population with high prevalence of gastric cancer. AB - AIM: To validate a non-invasive method to detect gastric mucosal atrophy in a Chilean population with high prevalence of gastric cancer and a poor survival rate. METHODS: We first determined the optimal cut-off level of serum pepsinogen (PG)-1, PG-1/PG-2 ratio and 17-gastrin in 31 voluntary symptomatic patients (mean age: 66.1 years), of them 61% had histologically confirmed gastric atrophy. Then, in a population-based sample of 536 healthy individuals (209 residents in counties with higher relative risk and 327 residents in counties with lower relative risk for gastric cancer), we measured serum anti-H pylori antibodies, PG and 17-gastrin and estimated their risk of gastric cancer. RESULTS: We found that serum PG-1 < 61.5 microg/L, PG-1/PG-2 ratio < 2.2 and 17-gastrin > 13.3 pmol/L had a high specificity (91%-100%) and a fair sensitivity (56%-78%) to detect corpus-predominant atrophy. Based on low serum PG-1 and PG-1/PG-2 ratio together as diagnostic criteria, 12.5% of the asymptomatic subjects had corpus-predominant atrophy (0% of those under 25 years and 20.2% over 65 years old). The frequency of gastric atrophy was similar (12% vs 13%) but H pylori infection rate was slightly higher (77% vs 71%) in the high-risk compared to the low-risk counties. Based on their estimated gastric cancer risk, individuals were classified as: low risk group (no H pylori infection and no atrophy; n = 115; 21.4%); moderate-risk group (H pylori infection but no atrophy; n = 354, 66.0%); and high-risk group (gastric atrophy, with or without H pylori infection; n = 67, 12.5%). The high risk group was significantly older (mean age: 61.9+/-13.3 years), more frequently men and less educated as compared with the low-risk group. CONCLUSION: We propose to concentrate on an upper gastrointestinal endoscopy for detection of early gastric cancer in the high-risk group. This intervention model could improve the poor prognosis of gastric cancer in Chile. PMID- 17131483 TI - Ultrasound guided percutaneous cholecystostomy in high-risk patients for surgical intervention. AB - AIM: To assess the efficacy and safety of ultrasound guided percutaneous cholecystostomy (PC) in the treatment of acute cholecystitis in a well-defined high risk patients under general anesthesia. METHODS: The data of 27 consecutive patients who underwent percutaneous transhepatic cholecystostomy for the management of acute cholecystitis from January 1999 to June 2003 was retrospectively evaluated. All of the patients had both clinical and sonographic signs of acute cholecystitis and had comorbid diseases. RESULTS: Ultrasound revealed gallbladder stones in 25 patients and acalculous cholecystitis in two patients. Cholecystostomy catheters were removed 14-32 d (mean 23 d) after the procedure in cases where complete regression of all symptoms was achieved. There were statistically significant reductions in leukocytosis, (13.7 x 10(3)+/-1.3 x 10(3) microg/L vs 13 x 10(3)+/-1 x 10(3) microg/L, P < 0.05 for 24 h after PC; 13.7 x 10(3)+/-1.3 x 10(3) microg/L vs 8.3 x 10(3)+/-1.2 x 10(3) microg/L, P < 0.0001 for 72 h after PC), C -reactive protein (51.2+/-18.5 mg/L vs 27.3+/-10.4 mg/L, P < 0.05 for 24 h after PC; 51.2+/-18.5 mg/L vs 5.4+/-1.5 mg/L, P < 0.0001 for 72 h after PC), and fever (38+/-0.35 centigrade vs 37.3+/-0.32 centigrade, P < 0.05 for 24 h after PC; 38+/-0.35 centigrade vs 36.9+/-0.15 centigrade, P < 0.0001 for 72 h after PC). Sphincterotomy and stone extraction was performed successfully with endoscopic retrograde cholangio-pancreatography (ERCP) in three patients. After cholecystostomy, 5 (18%) patients underwent delayed cholecystectomy without any complications. Three out of 22 patients were admitted with recurrent acute cholecystitis during the follow-up and recovered with medical treatment. Catheter dislodgement occurred in three patients spontaneously, and two of them were managed by reinsertion of the catheter. CONCLUSION: As an alternative to surgery, percutaneous cholecystostomy seems to be a safe method in critically ill patients with acute cholecystitis and can be performed with low mortality and morbidity. Delayed cholecystectomy and ERCP, if needed, can be performed after the acute period has been resolved by percutaneous cholecystostomy. PMID- 17131484 TI - Efficacy of leukocyte esterase dipstick test as a rapid test in diagnosis of spontaneous bacterial peritonitis. AB - AIM: To evaluate the efficacy of dipstick test in diagnosis of spontaneous bacterial peritonitis (SBP) in cirrhotic patients who underwent abdominal paracentesis based on the locally available dipstick test. METHODS: There were 200 consecutive samples from cirrhotic patients who underwent abdominal paracentesis. Urine dipstick (Combur10 TestM, Roche, Mannheim, Germany) was used as a screening test. A manual cell count with differential study was done in all samples by experienced technicians. The polymorphonuclear (PMN) cell count more than 250 cells/mm3 was used as a diagnostic cut off level. One to three plus dipstick results were used as cut off levels for a positive result. The dipstick test results had to be agreed by three experienced readers. The sensitivity, specificity, positive and negative predictive values and accuracy of two different colorimetric cut off scales (1+ and 2+) were calculated and compared. RESULTS: The prevalence of SBP diagnosed by manual cell count was 21.0%. There were 128 specimens that had a true negative result by dipstick. The sensitivity, specificity, positive and negative predictive values and accuracy of 1+ and 2+ cut off scale to diagnose SBP were 88%, 81%, 55%, 96% and 83% respectively, and 63%, 96%, 82%, 81% and 89% respectively. CONCLUSION: Dipstick test can be used as a rapid test for screening of SBP. The higher cut off colorimetric scale has a better specificity and positive predictive value but a lower sensitivity. PMID- 17131485 TI - Expression of altered retinoblastoma protein inversely correlates with tumor invasion in gastric carcinoma. AB - AIM: To investigate the clinical and pathological significance of altered retinoblastoma (Rb) encoding protein (pRb) in gastric carcinoma. METHODS: Expression of altered pRb was analyzed in 91 patients with gastric adenocarcinoma by immunohistochemistry. RESULTS: Sixty-five percent (59/91) of the tumors were positively stained and the staining in tumor nuclei of gastric carcinoma ranged 0%-90%. Moreover, strong expression of altered pRb was found in 35% (6/17), 24% (5/21), 17% (8/46) and 0% (0/7) of T1, T2, T3 and T4 gastric carcinomas, respectively. Altered pRb expression was inversely correlated with the depth of tumor invasion (P = 0.047). Degree of immunoreactivity had no significant correlation with tumor grade, node metastasis and distant metastasis. In terms of prognostic significance, univariate analysis showed that poor differentiation [41 (66.1%) vs 34 (42.5%) P = 0.051], advanced tumor stage (P < 0.001) and weakly altered pRb expression [17 (80.5%) vs 58 (49.6%) P=0.044] were associated with worse prognosis in these patients. However, multivariate analysis revealed that advanced tumor stage was the only independent poor prognostic factor (P < 0.001). CONCLUSION: the mutation of Rb gene is frequent in gastric carcinoma. The expression of altered pRb inversely correlates with tumor invasion and is not an independent prognostic marker in gastric adenocarcinoma. PMID- 17131486 TI - Comparison of amplicon-sequencing, pyrosequencing and real-time PCR for detection of YMDD mutants in patients with chronic hepatitis B. AB - AIM: To compare the sequencing of PCR products, pyrosequencing, and real-time PCR for detection of Tyrosine-methionine-aspartate-aspartate (YMDD) mutants in patients with chronic hepatitis B. METHODS: Mixtures of plasmids and serum samples from 69 chronic hepatitis B patients treated with lamivudine were tested for YMDD mutations by sequencing of PCR products, pyrosequencing, and real-time PCR, respectively. Time required and reagent costs of the three assays were evaluated. RESULTS: Real-time PCR detected 100%, 50%, 10%, 1% and 0.1% of YVDD plasmid in mixtures with 10(6) copies/mL of YMDD plasmid, whereas sequencing and pyrosequencing only detected 100% and 50% of YVDD plasmid in aliquots of the corresponding mixtures. Completely concordant results were obtained from 60 (87%) out of the 69 clinical serum samples by the three assays. Mutants were detected by real-time PCR in less than 20% of the total virus population, but no mutant was detected by sequencing and pyrosequencing. In addition, real-time PCR required less time and was more cost-effective than the other two assays. However, throughput of pyrosequencing was the highest. CONCLUSION: Among the three assays compared, real-time PCR is the most sensitive, cost-effective, and time saving for monitoring YMDD mutants in patients with chronic hepatitis B on lamivudine therapy. PMID- 17131487 TI - Hypertriglyceridemia-induced pancreatitis: A case-based review. AB - Hypertriglyceridemia is an established cause of pancreatitis. In a case-based approach, we present a review of hypertriglyceridemia and how it can cause pancreatitis. We outline how to investigate and manage such patients. A 35 year old man presented to the emergency department with abdominal pain and biochemical evidence of acute pancreatitis. There was no history of alcohol consumption and biliary imaging was normal. The only relevant past medical history was that of mild hyperlipidemia, treated with diet alone. Physical exam revealed epigastric tenderness, right lateral rectus palsy, lipemia retinalis, bitemporal hemianopsia and a delay in the relaxation phase of his ankle reflexes. Subsequent laboratory investigation revealed marked hypertriglyceridemia and panhypopituarism. An enhanced CT scan of the head revealed a large suprasellar mass impinging on the optic chiasm and hypothalamus. The patient was treated supportively; thyroid replacement and lipid lowering agents were started. He underwent a successful resection of a craniopharyngioma. Post-operatively, the patient did well on hormone replacement therapy. He has had no further attacks of pancreatitis. This case highlights many of the factors involved in the regulation of triglyceride metabolism. We review the common causes of hypertriglyceridemia and the proposed mechanisms resulting in pancreatitis. The incidence and management of hypertriglyceridemia-induced pancreatitis are also discussed. PMID- 17131488 TI - Endoscopic naso-pancreatic stent-guided single-branch resection of the pancreas for multiple intraductal papillary mucinous adenomas. AB - In benign or low-grade malignant pancreatic tumors, complete removal of the lesion is sufficient for a cure, and thus minimal resection techniques with preservation of the pancreatic functional reserve have advantages over more extended pancreatic resections. However, a high incidence of postoperative pancreatic fistula in such procedures has been reported. Moreover, branch-type intraductal papillary mucinous neoplasms of the pancreas tend to locate in the head of the pancreas, and show less malignant potential. We describe an endoscopic naso-pancreatic stent-guided single-branch resection of the pancreas for branch-type multiple intraductal papillary mucinous adenomas, along with a gastric wall-covering method for the prevention of pancreatic leakage. PMID- 17131489 TI - Thrombotic microangiopathy involving the gallbladder as an unusual manifestation of systemic lupus erythematosus and antiphospholipid syndrome: Case report and review of the literature. AB - Gallbladder disease is no more common in patients with systemic lupus erythematosus (SLE) than in the general population. We describe a 17-year-old patient with SLE, who developed nephritis that was well controlled with medications. Initial treatment consisted of azathioprine, aspirin and prednisone with stable control of her symptoms. Two years later she developed a right quadrant abdominal pain, and an abdominal ultrasound revealed microlithiasic cholecystitis. Open cholecystectomy was performed and the histopathological findings revealed vasculitis with thrombotic microangiopathy in the gallbladder. This case presentation illustrates that calculous or acalculous cholecystitis should be considered as a manifestation of active SLE and APS. PMID- 17131490 TI - Subcutaneous extension of a large diaphragmatic hydatid cyst. AB - A 53-year-old male patient with a large hydatid cyst of the left hemidiaphragm and smaller secondary cysts located in the left thoracic cavity and upper left abdominal quadrant presented with two progressively enlarging lipoma-like masses in the left hypochondrium and under the left scapulae respectively. Total excision of all the cysts was performed through a bilateral subcostal incision, with the left hemidiaphragm near totally excised and replaced by a synthetic bilayer mesh. PMID- 17131491 TI - Esophageal perforation caused by fish vertebra ingestion in a seven-month-old infant demanded surgical intervention: A case report. AB - A seven-month-old infant was admitted to our hospital with a 1-wk history of shortness of breath, dysphagia, and fever. Diagnosis of esophageal perforation following fish vertebra ingestion was made by history review, pneumomediastinum and an irregular hyperdense lesion noted in initial chest radiogram. Neck computed tomography (CT) confirmed that the foreign body located at the cricopharyngeal level and a small esophageal tracheal fistula was shown by esophagogram. The initial response to treatment of fish bone removal guided by panendoscopy and antibiotics administration was poor since pneumothorax plus empyema developed. Fortunately, the patient's condition finally improved after decortication, mediastinotomy and perforated esophagus repair. To our knowledge, this is the first case report of esophageal perforation due to fish bone ingestion in infancy. In addition to particular caution that has to be taken when feeding the innocent, young victim, it may indicate the importance of surgical intervention for complicated esophageal perforation in infancy. PMID- 17131492 TI - Synchronous occurrence of carcinoid, signet-ring cell carcinoma and heterotopic pancreatic tissue in stomach: A case report and literature review. AB - We presented an unusual case with coexistence of carcinoid, signet-ring cell carcinoma (SRC) and heterotopic pancreatic tissue in stomach. Gastroscopic examination of this 63-year-old male patient showed multiple protrusions in gastric corpus near the greater curvature, identified by subsequent biopsy as carcinoid. Distal subtotal gastrectomy was performed. Histological and immunohistochemical examinations showed a carcinoid tumor in gastric corpus near the greater curvature, an intramucosal SRC at the lesser curvature of corpus and heterotopic pancreatic tissue in muscularis propria of the antrum at the lesser curvature with hyperplasia of peripheral endocrine cells producing multiple pancreatic hormones. We reviewed the literatures on clinicopathological characteristics and the differential diagnosis of the above three abnormalities, and concluded that the carcinoid in corpus near the greater curvature and SRC in the lesser curvature are independent lesions; the foci of endocrine cells in the muscularis propria and serosa are hyperplastic lesions from the heterotopic pancreatic tissue, rather than dissemination of carcinoid in corpus. PMID- 17131493 TI - Hyperinsulinemic hypoglycemia due to adult nesidioblastosis in insulin-dependent diabetes. AB - In neonates, persistent hyperinsulinemic hypoglycemia (PHH) is associated with nesidioblastosis. In adults, PHH is usually caused by solitary benign insulinomas. We report on an adult patient who suffered from insulin-dependent diabetes mellitus, and subsequently developed PHH caused by diffuse nesidioblastosis. Mutations of the MEN1 and Mody (2/3) genes were ruled out. Preoperative diagnostic procedures, the histopathological criteria and the surgical treatment options of adult nesidioblastosis are discussed. So far only one similar case of adult nesidioblastosis subsequent to diabetes mellitus II has been reported in the literature. In case of conversion of diabetes into hyperinsulinemic hypoglycemia syndrome, nesidioblastosis in addition to insulinoma should be considered. PMID- 17131494 TI - Targeting intestinal microflora in inflammatory bowel disease. PMID- 17131495 TI - Premature chromosome condensation technique: A very promising approach to radiotherapy for digestive system cancers. PMID- 17131496 TI - Recent case law interpreting the Medicare Anti-Kickback Act. PMID- 17131497 TI - AHA adds Medicare, Medicaid. Move differs from CHA's on uncompensated care. PMID- 17131498 TI - "Texans are everywhere!". PMID- 17131505 TI - [Iscador and Helixor are not drugs!]. PMID- 17131506 TI - [Encourage the natural longing to give birth]. PMID- 17131507 TI - Last-minute battle. Election puts more uncertainty in fight against cuts. PMID- 17131509 TI - Termination of resuscitation in out-of-hospital cardiac arrest. PMID- 17131510 TI - Termination of resuscitation in out-of-hospital cardiac arrest. PMID- 17131511 TI - Termination of resuscitation in out-of-hospital cardiac arrest. PMID- 17131512 TI - Termination of resuscitation in out-of-hospital cardiac arrest. PMID- 17131513 TI - Termination of resuscitation in out-of-hospital cardiac arrest. PMID- 17131514 TI - Judging the safety of aprotinin. PMID- 17131515 TI - Injectable sustained-release risperidone: recurrence of delirium and treatment failure. PMID- 17131516 TI - Appetite suppressants and pulmonary hypertension: a long-term risk. PMID- 17131517 TI - [Medicine without scientific reasoning]. PMID- 17131518 TI - [Molecular genetics of myeloproliferative diseases]. PMID- 17131519 TI - [Medical treatment of Alzheimer's disease]. PMID- 17131520 TI - [Back to 1892?]. PMID- 17131521 TI - Correction for "Delayed brainstem auditory evoked potential latencies in 14-year old children exposed to methylmercury" by Murata et al. PMID- 17131522 TI - Minister pledges cash boost to Poland's health sector. PMID- 17131523 TI - China's cities get a little healthier. PMID- 17131524 TI - Bringing basic science back to centre stage to fight HIV/AIDS. PMID- 17131525 TI - Anger: heartbreaking at any age. PMID- 17131526 TI - Good nutrition: should guidelines differ for men and women? PMID- 17131527 TI - Massage therapy. PMID- 17131528 TI - On call. When I saw my doctor on my 73rd birthday, my only complaint was that I've been having trouble with my erections over the past year. I was surprised that my doctor didn't do any blood tests beyond the usual battery, and he didn't refer me to a specialist. Instead, he gave me a prescription for Viagra. The pills are working fine, but should I have tests to find out what's wrong? PMID- 17131529 TI - Refining the prescription for fish oil. Fish oil may not help those with implanted defibrillators. PMID- 17131530 TI - Central treatment for peripheral artery disease. Heart care is important for people with narrowed leg arteries. PMID- 17131531 TI - High blood pressure: common, commonly uncontrolled, and dangerous. 10 steps for fighting the good fight against high blood pressure. PMID- 17131532 TI - Policosanol: a sweet nothing for high cholesterol. PMID- 17131533 TI - Ask the doctor. Three years ago I had coronary bypass surgery. How long will my grafts last? PMID- 17131534 TI - Improving care for depression. New approaches to managing a chronic disease. PMID- 17131535 TI - Solution-focused therapy. PMID- 17131536 TI - Intermittent explosive disorder. PMID- 17131537 TI - The genetics of autism. PMID- 17131538 TI - Why drug addicts die. PMID- 17131539 TI - Questions & answers. Is anorexia ever a problem for men? PMID- 17131540 TI - [Eating disorders in children]. PMID- 17131541 TI - [History of social phobia and social anxiety disorder]. PMID- 17131542 TI - [Morita therapy for social anxiety disorders]. PMID- 17131543 TI - [Therapy for social anxiety spectrum]. PMID- 17131545 TI - Foie gras and contemporary animal welfare. PMID- 17131544 TI - Molecular and genetic markers in thoracic surgery. PMID- 17131546 TI - Abstracts of the 16th Annual Meeting of the German Society for Cytometry, 18-21 October 2006, Leipzig, Germany. PMID- 17131547 TI - [Cognitive disorders]. PMID- 17131548 TI - Selected clinical trials in breast cancer. PMID- 17131549 TI - [Is it necessary to elimate the voices?]. PMID- 17131550 TI - [Is it necessary to eliminate the voices? That is not the question! Response by a former hearer of voices]. PMID- 17131551 TI - [Is it necessary to elimate the voices? A response]. PMID- 17131552 TI - [Interior prison or phantom discourse?]. PMID- 17131553 TI - [Voices and hallucinations: some lessons and pitfalls]. PMID- 17131554 TI - [The risk of losing one's "voice" or the dangers of loss of the mind in psychiatry]. PMID- 17131555 TI - [Voices, symptoms to interpret before deciding to treat them]. PMID- 17131556 TI - [Hallucinatory perceptions]. PMID- 17131558 TI - Notice of duplicate publication. PMID- 17131557 TI - [Proceedings of the 28th International Congress of the International Commission on Occupational Health, Milan, Italy, 11-16 June 2006]. PMID- 17131559 TI - Relatives' perspective on the terminally ill patients who died after euthanasia or physician-assisted suicide: a retrospective cross-sectional interview study in the Netherlands. AB - This study used retrospective interviews with 87 relatives to describe the experiences of patients who died by euthanasia or physician-assisted suicide (EAS) in the Netherlands. Most of the patients suffered from cancer (85%). The relatives were most often a partner (63%) or a child (28%) of the patient. Before explicitly requesting EAS most patients (79%) had spoken about their wishes concerning medical end-of-life decisions to be made at a later date. Hopeless suffering, loss of dignity, and no prospect of recovery were the most prevalent reasons for explicitly requesting EAS. According to the relative, in 92% of patients EAS had contributed favourably to the quality of the end of life, mainly by preventing or ending suffering. PMID- 17131560 TI - The influence of maternal loss on young women's experience of identity development in emerging adulthood. AB - A qualitative approach was used to study the influence of adolescent maternal loss on identity development in 6 young women. In-depth, semi-structured interviews were audiotaped, transcribed, and analyzed for recurrent themes. Three metathemes emerged from the narratives: Loss of Mother Impacts Identity, Relatedness on the Path of Identity Development, and Integration of Loss into Emergent Identity. Findings indicate that the experience of maternal loss influenced aspects of the women's identity development and initiated a need to seek new connections and redefine aspects of self in order to integrate the loss. Practical implications of these findings are presented for clinicians who work with bereaved young women. PMID- 17131561 TI - Suicidality and migration among adolescents in Hong Kong. AB - Suicide as a cause of death among adolescents and migration as a component of population have been growing in importance. Very little research has been conducted on the connections between migration and suicidality among adolescents in Hong Kong, and so is the aim of this article. It uses census and registration data to study suicide mortality, and sample survey data collected for this purpose to investigate suicide attempt, suicide ideation, and self-injurious behavior. Relations between suicidality and socio-demographic/psychological factors replicated those found in the literature. Duration of residence was found important for the study of suicide among migrants. In both the bivariate and multivariate analyses, although the suicidality levels for short-duration (less than 10 years) adolescent migrants were very much lower than the local-born counterparts, those for the long-duration (10 years or more) migrants were very much higher. The findings support the Healthy Migrant Hypothesis and other related hypotheses in migrant mortality studies. They also reveal, in the light of the Integration Theory of Suicide, the problem of migrant integration into the host culture and society, an important social problem for the government to solve. PMID- 17131562 TI - South African Zulu widows in a time of poverty and social change. AB - Interviews were carried out with 16 South African Zulu widows. Much of what the widows had to say seemed like what one might hear from widows in economically developed countries, but there were also striking differences. All the widows lived in poverty, and for some their grief seemed much more about the poverty than about the husband's death. Most widows observed a year of traditional ukuzila mourning practices, which made them a threat to others and which is difficult to carry out in the social upheaval of modern South Africa. Widowhood was in some cases a struggle with witchcraft--as cause of the death or as an accusation directed at the widow. PMID- 17131563 TI - Children who lost a parent as a result of the terrorist attacks of September 11, 2001: registry construction and population description. AB - Children who experience traumatic bereavement in the context of catastrophic disasters are at increased risk for developing post-disaster problems. Despite massive loss of life on September 11th, 2001, no public data were collected on those children who lost a parent in the multiple terrorist attacks. Such a registry would be an important public health tool to help mitigate the consequences of traumatic bereavement and to guide health surveillance efforts. As information about affected individuals was fragmented and difficult to congregate, we constructed our registry by conducting systematic public record searches and data exchanges with key stakeholders to identify, locate, and characterize all the 9/11 bereaved children. We identified 1,363 victims who were parents of minor children, and 2,752 bereaved children below age 18. In addition to reporting important identifying data on these children, this study also demonstrates the feasibility and the potential usefulness of such a registry to support post-disaster service provision and advocacy efforts. The construction of registries of severely affected people post-disaster should become government policy to obviate the need to use ad hoc methods to construct lists of high-risk individuals. PMID- 17131564 TI - Innovation: the classic traps. AB - Never a fad, but always in or out of fashion, innovation gets rediscovered as a growth enabler every half dozen years. Too often, though, grand declarations about innovation are followed by mediocre execution that produces anemic results, and innovation groups are quietly disbanded in cost-cutting drives. Each managerial generation embarks on the same enthusiastic quest for the next new thing. And each generation faces the same vexing challenges- most of which stem from the tensions between protecting existing revenue streams critical to current success and supporting new concepts that may be crucial to future success. In this article, Harvard Business School professor Rosabeth Moss Kanter reflects on the four major waves of innovation enthusiasm she's observed over the past 25 years. She describes the classic mistakes companies make in innovation strategy, process, structure, and skills assessment, illustrating her points with a plethora of real-world examples--including AT&T Worldnet, Timberland, and Ocean Spray. A typical strategic blunder is when managers set their hurdles too high or limit the scope of their innovation efforts. Quaker Oats, for instance, was so busy in the 1990s making minor tweaks to its product formulas that it missed larger opportunities in distribution. A common process mistake is when managers strangle innovation efforts with the same rigid planning, budgeting, and reviewing approaches they use in their existing businesses--thereby discouraging people from adapting as circumstances warrant. Companies must be careful how they structure fledgling entities alongside existing ones, Kanter says, to avoid a clash of cultures and agendas--which Arrow Electronics experienced in its attempts to create an online venture. Finally, companies commonly undervalue and underinvest in the human side of innovation--for instance, promoting individuals out of innovation teams long before their efforts can pay off. Kanter offers practical advice for avoiding these traps. PMID- 17131565 TI - Managing multicultural teams. AB - Multicultural teams offer a number of advantages to international firms, including deep knowledge of different product markets, culturally sensitive customer service, and 24-hour work rotations. But those advantages may be outweighed by problems stemming from cultural differences, which can seriously impair the effectiveness of a team or even bring itto a stalemate. How can managers best cope with culture-based challenges? The authors conducted in-depth interviews with managers and members of multicultural teams from all over the world. Drawing on their extensive research on dispute resolution and teamwork and those interviews, they identify four problem categories that can create barriers to a team's success: direct versus indirect communication, trouble with accents and fluency, differing attitudes toward hierarchy and authority, and conflicting norms for decision making. If a manager--or a team member--can pinpoint the root cause of the problem, he or she is likelier to select an appropriate strategy for solving it. The most successful teams and managers, the authors found, dealt with multicultural challenges in one of four ways: adaptation (acknowledging cultural gaps openly and working around them), structural intervention (changing the shape or makeup of the team), managerial intervention (setting norms early or bringing in a higher-level manager), and exit (removing a team member when other options have failed). Which strategy is best depends on the particular circumstances--and each has potential complications. In general, though, managers who intervene early and set norms; teams and managers who try to engage everyone on the team; and teams that can see challenges as stemming from culture, not personality, succeed in solving culture-based problems with good humor and creativity. They are the likeliest to harvest the benefits inherent in multicultural teams. PMID- 17131566 TI - Breaking the trade-off between efficiency and service. AB - For manufacturers, customers are the open wallets at the end of the supply chain. But for most service businesses, they are key inputs to the production process. Customers introduce tremendous variability to that process, but they also complain about any lack of consistency and don't care about the company's profit agenda. Managing customer-introduced variability, the author argues, is a central challenge for service companies. The first step is to diagnose which type of variability is causing mischief: Customers may arrive at different times, request different kinds of service, possess different capabilities, make varying degrees of effort, and have different personal preferences. Should companies accommodate variability or reduce it? Accommodation often involves asking employees to compensate for the variations among customers--a potentially costly solution. Reduction often means offering a limited menu of options, which may drive customers away. Some companies have learned to deal with customer-introduced variability without damaging either their operating environments or customers' service experiences. Starbucks, for example, handles capability variability among its customers by teaching them the correct ordering protocol. Dell deals with arrival and request variability in its high-end server business by outsourcing customer service while staying in close touch with customers to discuss their needs and assess their experiences with third-party providers. The effective management of variability often requires a company to influence customers' behavior. Managers attempting that kind of intervention can follow a three-step process: diagnosing the behavioral problem, designing an operating role for customers that creates new value for both parties, and testing and refining approaches for influencing behavior. PMID- 17131567 TI - Facing ambiguous threats. AB - On February 1, 2003, the world watched in horror as the Columbia space shuttle broke apart while reentering the earth's atmosphere, killing all seven astronauts. Some have argued that NASA's failure to respond with appropriate intensity to the so-called foam strike that led to the accident was evidence of irresponsible or incompetent management. The authors' research, however, suggests that NASA was exhibiting a natural, albeit unfortunate, pattern of behavior common in many organizations. The foam strike is a prime example of what the authors call an ambiguous threat-a signal that may or may not portend future harm. Ambiguous threats differ from threats with obvious causes-say, a fire in the building-for which the response is clear. They also differ from unmistakable threats that may lack straightforward response paths (such as the frightening oxygen-tank explosion aboard Apollo 13). However, when the warning sign is ambiguous and the threat's potential effect is unclear, managers may choose to ignore or discount the risk. Such an approach can be catastrophic. Firms that do a good job of dealing with ambiguous threats do not improvise during a crisis; rather, they apply a rigorous set of detection and response capabilities that they have developed and practiced beforehand. In this article, the authors outline how to put such capabilities in place long before a crisis strikes. First, companies need to hone their teamwork and rapid problem-solving skills through practice. Second, they must learn to recognize weak signals, amplify the threat, and encourage employees to ask disconcerting "what if" questions in a safe environment. Finally, they should explore possible responses to threats through quick, low-cost experimentation. PMID- 17131568 TI - Disaster relief, inc. AB - When disaster strikes, many corporations respond generously. After the 2004 tsunami, for instance, U.S. firms alone contributed more than half a billion dollars in cash and in-kind donations. But a host of reactive efforts don't produce the best results-and may even get in the way. To make the most of their humanitarian efforts, companies need to address two fundamental questions: What kind of aid do we want to contribute--philanthropic (money and in-kind donations) or integrative (backroom, operational assistance)? And how do we want to contribute it--by working one-on-one with a single agency or by joining a consortium? The permutations of those two decisions lead to four different approaches, each with its own strengths and challenges. Single-company philanthropic partnerships work well when there's a good match between what a company wants to contribute and what an agency needs, as with Coca-Cola's donations of water to the Red Cross. More diffuse, but also potentially more effective, are the benefits of joining a multicompany philanthropic partnership, which enables the resources of many firms to be matched to the missions of many agencies. More difficult to establish but more fundamental in its impact is a single-company integrative partnership, in which a corporation works to improve the way an aid agency operates, as the logistics giant TNT has done to help the distribution efforts of the World Food Programme. And most difficult to implement -but potentially most effective-is a multicompany integrative partnership, which brings to bear the collective best practices of many companies to improve the response capabilities of multiple agencies. It's easy to see why the image of a relief worker carrying a sack of grain delivers an emotional wallop, but the behind-the-scenes work of process enhancement is just as crucial to humanitarian efforts. The sooner executives realize this, the better positioned the world will be to respond to global disasters. PMID- 17131569 TI - How well-run boards make decisions. AB - In the aftermath of seismic debacles like those that toppled Enron and WorldCom, corporate boards have been shaken up and made over. More directors are independent these days, for instance, and corporations now disclose directors' salaries and committee members' names. Research shows that most of the changes are having a positive effect on companies' performance. They are primarily structural, though, and don't go to the heart of a board's work: making the choices that shape a firm's future. Which decisions boards own and how those calls are made are largely hidden from the public. As a result, boards are often unable to learn from the best governance practices of their counterparts at other companies. This article pulls back the curtain and provides an inside look. Drawing on interviews with board members and executives at 31 companies, along with a close examination of three boardroom decisions, the author identifies several formal processes that can help companies improve their decision making: creating calendars that specify when the board and the standing committees will consider key items; drafting charters that define the decisions committees are responsible for; and developing decision protocols that divvy up responsibilities between directors and executives. The author also identifies a number of informal decision-making principles: Items that are strategically significant and touch on the firm's core values should go to the board. Large decisions should be divided into small pieces, so the board can devote sufficient attention to each one. Directors must remain vigilant to ensure that their decisions are effectively implemented. The CEO and either the nonexecutive chair or the lead director should engage in ongoing dialogue regarding which decisions to take to the full board and when. And directors should challenge assumptions before making yes-or no decisions on management proposals. PMID- 17131570 TI - From Kerala to Kyrgyzstan. PMID- 17131571 TI - This life. PMID- 17131572 TI - Housing and community care. PMID- 17131573 TI - Made to measure? PMID- 17131574 TI - Woman on the verge. PMID- 17131575 TI - Internet on psychiatric wards. PMID- 17131576 TI - Avoiding protests and picket lines: staying union free. PMID- 17131577 TI - GHA should lead the way in transparency efforts. PMID- 17131578 TI - Georgia medical malpractice release authorization preempted by HIPAA. PMID- 17131579 TI - Putting an end to judicial activism. PMID- 17131580 TI - "A rough guide to the mind" a Festschrift in honour of Prof. John C. Marshall. PMID- 17131581 TI - Now you see it, now you don't: on turning semantic interference into facilitation in a Stroop-like task. AB - We use a masked priming procedure to test two accounts of the picture-word interference (PWI) effect: the lexical selection by competition account (Levelt et al., 1999; Roelofs, 1992) and the response selection account (Lupker, 1979; Miozzo and Caramazza, 2003). In the visible (standard) condition, we replicated the often-observed semantic interference effect. In the masked condition, we observed semantic facilitation. We take the polarity shift as a function of masking to mean that the semantic interference and semantic facilitation in the PWI task should be attributed to two qualitatively different processes. We argue that this conclusion follows naturally from the response selection account, but only with great difficulty from the lexical selection by competition account. PMID- 17131582 TI - Aphasic deficits in syntactic processing. AB - Two main classes of theories of deficits in syntactic processing in aphasia have been suggested: impairments affecting specific syntactic operations and reductions in resources available for syntactic processing. This paper reviews the data upon which such theories are based and argues that they may support only the latter type of model. PMID- 17131583 TI - A linguist's views on progressive anomia: evidence for Delbruck (1886) in modern neurolinguistic research. AB - In his short paper of 1886, the neogrammarian linguist Delbruck sketches his views on normal language processing and their relevance for the interpretation of some of the symptoms of progressive anomic aphasia. In particular, he discusses proper name impairments, verb and abstract noun superiority and the predominance of semantically related errors. Furthermore, he suggests that part of speech, morphology and word order may be preserved in this condition. This historical document has been lost in oblivion but the original ideas and their relevance for contemporary discussions merit a revival. PMID- 17131584 TI - Meynert on Wernicke's aphasia. AB - This paper examines Meynert's contribution to aphasia, in particular the suggestion that Meynert already had described the syndrome of sensory aphasia. I examine Meynert's own writings on this subject, Wernicke's statements on this issue and biographies of Meynert and Wernicke. I argue that Meynert did not describe sensory aphasia, nor is there convincing evidence that he stimulated Wernicke in this direction. Meynert was primarily interested in the global neuroanatomical organization of the brain and in particular the special role of the frontal lobes. PMID- 17131585 TI - Word finding in the damaged brain: probing Marshall's caveat. AB - Marshall (1977) constructed a plausible simulation of "anomic" speech out of the 100 most common words in the English language. He suggested that impaired access to lower frequency vocabulary might underlie anomic word finding difficulties. But he also noted that another factor, age of acquisition, may exert an influence, with anomic patients experiencing particular difficulty with later acquired vocabulary. A review of research on word-finding in aphasia and other neuropsychological conditions suggests that Marshall (1977) may have been right on both counts, and that in many patients both frequency of use and age of acquisition influence the likelihood that a given word will be able to be accessed and used. Theoretical accounts of why the age of acquisition of words might affect their retention or loss following brain injury in adulthood are considered. PMID- 17131586 TI - Emotional words: what's so different from just words? AB - Aphasic patients, in particular global aphasics, may still swear and produce emotional utterances with ease. Based on these clinical observations we investigated emotional word "reading" in a series of different experiments over 25 years, not only in aphasic patients, but also in the left (LVF) and right (RVF) visual fields of healthy subjects, and in a depth-recorded epileptic patient. Across these experiments we found: i) behaviorally a strong emotional word effect in the left visual field (right hemisphere - RH) of normals, correlating well with the emotional word performance of aphasic patients, pointing to a specific role of the right hemisphere; ii) electro-physiologically a specific early (100-140 msec) brain response to emotional words during scalp recordings in healthy subjects subsequent to right visual field (left hemisphere LH) stimulation, that source localization procedures project to posterior areas of the right hemisphere; iii) preliminary data of a very early (60 msec) activation of the left amygdala in a depth-recorded epileptic patient when the same emotional words were presented to the right visual field (left hemisphere); and iv) a consistent gender difference showing that the above findings might be relevant for men only. Both hemispheres therefore appear to be implicated in emotional word "reading" but in different ways. We propose that the left amygdala via extrastriate connections acts as a detector of emotional word content at a very early stage of processing; that this amygdala response subsequently modulates the cortical response to emotional words asymmetrically, rendering the left visual cortex less sensitive to emotional words than that of the right hemisphere; and that this modulation is gender dependent. PMID- 17131587 TI - Jumping to conclusions: "language loss" versus "language impairment". AB - This paper discusses the impact of different metaphors of language dysfunction, with particular notice taken of the contribution of Marshall's work. This is especially the case regarding the notable paper on semantic error patterns (Marshall and Newcombe, 1973), and his later commentary on Chomsky's Rules and Representations (1980) with his elaboration of the conceptual commitments of The New Organology (1980). Examples of language performance provided by two sets of studies are used to illustrate the view that talk of language "loss" is hard to differentiate from the effects of concomitant processing mechanisms such as working memory. These examples are taken from studies of English and Turkish dementia of the Alzheimer type (DAT) patients (March, 2004), and temporal lobe epilepsy (TLE) patients (Field, 2001). Much contemporary neuropsychological research seems focussed on issues which assume that we already know how to describe the behaviours and processes we are interested in, and thus we can now concentrate on how and where in the brain these processes are handled. This perspective is examined from the points of view both that we do not yet have a sufficiently precise application of descriptions which utilise Linguistic constructs, and also that the available models of such processes as memory do not yet allow us to interpret task effects independently of the theoretician's intuitions. PMID- 17131588 TI - The cognitive genetics of attention deficit hyperactivity disorder (ADHD): sustained attention as a candidate phenotype. AB - Here we describe the application of cognitive genetics to the study of attention deficit hyperactivity disorder (ADHD). Cognitive genetics owes much to the pioneering work of cognitive neuropsychologists such as John Marshall, whose careful observations of cognitive dissociations between brain-lesioned patients greatly advanced the theoretical understanding of normal cognitive function. These theories have in turn helped to constrain linkages between candidate genes and cognitive processes and thus help to drive the relatively new field of cognitive genetics in a hypothesis-driven fashion. We examined the relationship between sustained attention deficits in ADHD and genetic variation in a catecholamine-related gene, dopamine beta hydroxylase (DbetaH). DBH encodes the enzyme that converts dopamine to noradrenaline and is crucial to catecholamine regulation. A polymorphism with the DBH gene has been associated with ADHD. In fifty-two children with ADHD, we examined whether variation in the Taq I DBH gene polymorphism was related to sustained attention performance. Participants performed the Sustained Attention to Response Test (SART). Performance on the SART discriminates ADHD from control children, and in imaging work, is associated with right frontoparietal activation. A significant effect of DBH genotype was found on SART performance measures. Children possessing two copies of the ADHD associated risk allele (A2) had significantly poorer sustained attention than those ADHD children who did not possess this allele or a non-genotyped control group. The DBH gene may contribute to the susceptibility for ADHD, in part because of its varying effects on the development of brain mechanisms mediating sustained attention. PMID- 17131589 TI - The "soul" of language does not use statistics: reflections on vowels and consonants. AB - This paper reviews studies of language processing with the aim of establishing whether any type of statistical information embedded in linguistic signals can be exploited by the language learner. The constraints as to the information that can be so used, we will argue, should be used to inform theories of language acquisition. We present two experiments with their respective controls. Both show that consonants (Cs) are much more suitable than vowels (Vs) to parse speech streams using statistical dependencies. These experiments use streams composed of items in which statistical information is carried either by the sequence of consonants or by the sequence of vowels. Both kinds of items are simultaneously present is the speech stream but, crucially, their overlap is only partial. Since the location of dips in transitional probabilities (TPs) between adjacent syllables differ for the first and the second type of sequences, we can explore whether consonants and vowels are equally efficient segments to parse signals. Our results show that "consonant words" (CW) are significantly preferred over "vowel words" (VW). We discuss the implication of our results for models of language acquisition. PMID- 17131590 TI - John Marshall and the cognitive neuropsychology of reading. AB - Contemporary cognitive neuropsychology owes a great debt to John Marshall. His pioneering work on acquired and developmental disorders of reading established the principles by which cognitive-neuropsychological theorizing operates as well as leading to a profound understanding of the nature of reading disorders which has subsequently influenced their assessment and treatment. PMID- 17131591 TI - Unilateral neglect is not unilateral: evidence for additional neglect of extreme right space. AB - Six patients with visuospatial neglect following right hemisphere lesions were given three tasks that assessed performance in areas of space ranging from extreme left to extreme right. A line bisection task required the patients to detect and bisect lines of four different lengths at seven left-right spatial locations, a number report task required the patients to name 11 two-digit numbers in a left-right array, and a tiling task required patients to place small black tiles over the black squares of a grid that stretched from 65 degrees left to 65 degrees right. Performance was compared with that of 20 age-matched controls. The patients showed the characteristic signs of left-side neglect in left space, extending to the central midline. Performance was relatively normal in centre-right space but all 6 patients showed signs of neglect of extreme right space (60 degrees to the right of the midline and beyond). We propose that neglect is best characterised as a bilateral, asymmetrical compression of experienced space in which the constriction extends further from the left than from the right but nevertheless affects both sides of space. PMID- 17131592 TI - From unilateral neglect to the brain basis of consciousness. AB - According to the dominant focus model, the differential activation of the thalamocortical network is a sufficient condition for the diversity of states of consciousness to occur, and coherent patterns of interactive forebrain activation peaks are sufficient to generate specific conscious figures against ground. These minimal assumptions are consonant with the known properties of unilateral neglect. It becomes unnecessary to suppose that consciousness is an independent entity, occupies a dedicated location in the brain, is contingent on a minimal level of neural complexity, plays a causal role in enabling perception or action, or evolved because it plays any such role. PMID- 17131593 TI - Verb-noun double dissociation in aphasia: theoretical and neuroanatomical foundations. AB - This paper reports the results of several studies on the mechanisms underlying Verb-Noun (V-N) dissociation. The objectives of the studies were to ascertain the location of the lesions causing predominant V or N impairment and to shed light on the different mental representations of these word classes through analyses of the data from neuropsychological patients. With regard to lesion sites, results obtained through an anatomo-correlative study on 15 V-impaired and 5 N-impaired aphasic patients indicate that lesions causing predominant N impairment were mostly located in the middle and inferior left temporal area. Three alternative lesion sites were associated with a V deficit (left posterior temporo-parietal lesions; large left fronto-temporal perisylvian lesions; deep lesions of the insula and/or the basal ganglia). In contrast to the results obtained from several neuropsychological and neuroimaging studies, none of the V-impaired patients had an isolated frontal lesion. The second aim is to discuss grammatical class interaction with semantic factors such as actionality or imageability (said to be the real cause of V-N dissociation). The retrieval of Ns and Vs in a sentence context was tested on 16 V-impaired aphasic patients and the resulting data indicate that imageability interacts with the retrieval of Ns and Vs, but cannot completely account for their dissociation. PMID- 17131594 TI - Retrieval pathways for common and proper names. AB - Paradigmatic cases of proper name anomia and proper name selective sparing are reviewed from relevant neuropsychological literature. Available evidence supports the existence of functionally and anatomically distinct retrieval pathways for the categories of proper and common names. An information processing model whose main feature lies in the relative independence within the semantic-conceptual system of information concerning individual entities may account for most of the observed phenomena. Localization studies seem to indicate that a complex neural network sustains various tasks implied in proper name processing. A dedicated module dealing with proper name retrieval is probably there, but its location within the left hemisphere is not at present fully understood. The proper name specific retrieval process is shown, in keeping with current philosophical and linguistic theories, to be intrinsically fragile and source-consuming. PMID- 17131595 TI - Patterns of peripheral paralexia: pure alexia and the forgotten visual dyslexia? AB - The concept of visual dyslexia put forward by Marshall and Newcombe (1973) is assessed. After a long period of neglect it was resurrected in the late 1990s in a narrow form. In the current paper it is proposed that a wider form of the functional syndrome is useful to include amongst other conditions attentional dyslexia and neglect dyslexia. The variety of sub-forms would correspond to the behavioural effects of the different ways in which the orthographic processing systems can be impaired. What distinguishes the broader form from pure alexia is that the patient lacks the capacity to use a serial letter processing strategy, and so interpretation of visual dyslexia in terms of the impairment to the orthographic processing systems is not contaminated by the use of a compensatory strategy that results in processing operations which are qualitatively very different from the normal and highly opaque. The lack of a serial letter processing strategy makes visual dyslexia a much more transparent functional syndrome. PMID- 17131596 TI - Developmental and acquired dyslexias. AB - Marshall (1984) highlighted potential parallels between children with developmental disorders of reading and adults who had acquired reading disorders. He advocated the use of a cognitive neuropsychological framework in the investigation of children with developmental abnormalities of cognition, including those with developmental dyslexias. Developmental phonological dyslexia has been extensively described and is a pervasive disorder. The relationship between reading difficulty and phonological difficulties evident in explicit oral phonological tasks continues to be a focus for debate. Clear cases of developmental deep dyslexia have now been described and the syndrome has also been described as characterising early reading development in Williams syndrome (WS), where there are also semantic errors in other domains, including naming and receptive vocabulary and there may be a generalised difficulty with the activation of fine grain semantic specifications. In the domain of number, highly selective reading disorders characterised by high rates of semantic errors have been documented, indicating that semantic reading errors can be domain-specific. They can occur to number words despite intact ability to read Arabic numbers and they can occur to Arabic numbers and number words despite intact ability to read words in other domains. Current models of reading written words do not allow for such material-specific dissociation. Developmental surface dyslexia has also been described in a range of countries, languages and orthographies. Descriptions of cases for whom there is no phonological impairment in reading have generated contrary evidence for theories suggesting that phonological impairment underlies all developmental dyslexia. As reading develops in Williams Syndrome, phonological reading skills may improve with over-reliance on these leading to surface dyslexia. Surface dyslexia has also been reported in cases of developmental amnesia in which there are semantic memory impairments. Hyperlexia can take several forms including broad hyperdevelopment with elevated phonological reading abilities, lexico-semantic reading abilities and reading comprehension as in Turner's syndrome (TS). This advantage has early onset in school-starters. These specific modular effects do not have pervasive impact across systems but demonstrate the limitations of functional plasticity in developmental and genetic disorders. The framework Marshall (1984) outlined has provided a foundation for the development of systematic investigation of developmental disorders. PMID- 17131597 TI - Productive and defective impairments in the neglect syndrome: graphic perseveration, drawing productions and optic prism exposure. AB - The effects of adaptation to prisms displacing rightwards the field of vision on omission errors, and on perseveration and other graphic productions in a line cancellation task, were assessed in nine right-brain-damaged patients with left unilateral spatial neglect. Prism adaptation improved both neglect, as indexed by omission errors, and perseveration behaviour, up to a delay of 60 min. No correlation was found between omission and perseveration errors in all assessments. The suggestion is made that perseveration and other complex graphic productions made by right brain-damaged-patients with left spatial unilateral neglect is due to a defective monitoring of complex motor behaviour, frequently associated to cerebral damage involving the right frontal lobe. Interpretations of perseveration behaviour in terms of allochiria and directional hypokinesia are considered, and their limits discussed. PMID- 17131598 TI - A method for studies of madness. AB - In these series of articles honouring John Marshall for his contributions to psychology, linguistics, the history of ideas and many other fields we should not forget his contribution to psychiatry. This has been mostly from the point of view of a critical observer, but also someone deeply interested in the application of cognitive neuropsychology to a broader range of disorders than the traditional ones. This "cognitive neuropsychiatric approach" is discussed in relation to attempts to understand thought disorder, delusions and lack of insight. PMID- 17131599 TI - When Ciarli Ciaplin drives a Pejio' in Itali: writing loan words in a shallow language. AB - With increasing frequency the lexicon of every language incorporates words taken from other languages (loan words) that become of common use, but whose orthography does not necessarily conform to the rules of the native language. This is particularly true for languages like Italian, where most words can be correctly written by the application of a sublexical phonology-to-orthography conversion procedure. Here we report the case of a neurologically unimpaired, highly educated Italian person, with a specific deficit in writing loan words. The aim of the study is to clarify the nature of her dysgraphic impairment, if linked to a semantic deficit, or alternatively reflecting an acquisition deficit, present both in the phonological and orthographic output lexicons, or specific only for the orthographic lexicon. A qualitative analysis of her errors and a comparison with the performance of a group of age and education matched subjects showed that her error pattern was not an amplification of the task difficulty effect observed in the control subjects, but rather an expression of an impairment in acquiring specific writing procedures. PMID- 17131600 TI - Hand preference and performance in 20 pairs of monozygotic twins with discordant handedness. AB - The differences between right (RH) and left (LH) handers reported in the literature on fine motor tasks, has traditionally been interpreted relative to purported functioning of the cerebral hemispheres. However, conclusive evidence for performance differences which are intrinsic to handedness per se is difficult to obtain unless left and right handers are compared who are similar in their genetic and environmental background. The present study therefore, employed a monozygotic (MZ) twin design which minimizes differences in genetic variation between the two groups. Forty female monozygotic twins (20 pairs) were selected on the basis of discordance of writing hand. Their laterality preferences were assessed and they were tested for differences on hand performance tasks (dot filling, finger tapping, and peg moving). The results revealed that on the hand and foot preference inventories, the right-handers were more strongly lateralized than their left-handed sisters, and that the left-handers had greater variation in their laterality scores. There were substantial correlations between preference and performance scores. The analyses not only revealed the obvious strong main effects of writing hand on performance tasks, but interaction effects of handedness on the peg-moving task. The dot filling task differentiated the writing versus non-writing hand considerably better than either of the other two performance tasks. However, no evidence was found to indicate that twins who wrote with their left hands showed poorer performance than their right-handed twin sisters. PMID- 17131601 TI - Re-visiting "semantic facilitation" of word retrieval for people with aphasia: facilitation yes but semantic no. AB - Previous research has shown that word-to-picture matching for targets that cannot be named at pre-test results in improved naming relative to untreated control items for people with aphasia. This paper replicates and extends this finding and investigates its source. Is the effect a result of priming of semantic representations, or of post-semantic mechanisms in word retrieval? The first experiment shows that word-to-picture matching with unrelated distractors improves naming at short (2-3 minutes) and long (up to 25 minute) lags. There was no effect of being made aware of the relationship between word-to-picture matching and picture naming. People who have a semantic impairment improve only with a short lag between word-to-picture matching and naming. Participants with less semantic impairment show larger priming effects that are equal at short and long lags between word-to-picture matching and naming. The second experiment shows that the facilitation effect is just as large for word-to-picture matching with unrelated distractors as with semantically-related distractors. Furthermore, overall there was no difference between matching with coordinate items and with associated items. The results of these experiments show that facilitation of naming by word-to-picture matching in people with aphasia cannot be a result of the priming of semantic representations. Instead they are consistent with two effects: word-to-picture matching results in priming at a lemma level for aphasic people with a semantic impairment that is only found with a short lag between word-to-picture matching and naming. Word-to-picture matching causes priming of the lemma to output lexicon entry mapping that benefits participants with less semantic impairment that is evident at both a short and long lag between word-to picture matching and naming. These findings fit well with previous research on repetition priming of naming with normal subjects. PMID- 17131602 TI - Sex differences in neural mechanisms underlying implicit symmetry processing. AB - Some species, including humans, prefer symmetrical to asymmetrical patterns. Preferences for symmetrical stimuli could have arisen in both sexes because specialized systems developed to detect and identify symmetrical stimuli, which are more likely to be figure than background. Females, it has been suggested, prefer symmetry because it may provide a cue to biological fitness when choosing a mate. Using functional magnetic resonance imaging, we found sex differences in neural activity when subjects distinguished figure from ground in abstract patterns whose component shapes differed in colour, brightness, and symmetry/asymmetry. Although no participant explicitly reported using symmetry as a decision criterion, women showed more neural activity than men in visual processing areas when the display contained a symmetrical shape. This enhanced activity in occipital cortex (middle and superior occipital gyrus) occurred bilaterally in women irrespective of whether they chose the symmetrical element in the display as the figure. Contrariwise, men showed a significant neural response in right temporal (superior temporal gyrus) and left parietal cortex (inferior parietal cortex/temporo-parietal junction) only when they chose a symmetrical element as the figure. The female brain thus appears to register symmetry automatically as a stimulus attribute, while the differential neurophysiological response of the male brain is dependent upon an explicit behavioural response to symmetry (choosing the symmetrical part of the display as figure), even if the criterion for choice (symmetry) is not reported verbally. PMID- 17131603 TI - Have we achieved gender equality? PMID- 17131604 TI - A gendered study of young adult contraceptive use at one university in KwaZulu Natal. AB - This study explores contraceptive use among young adult male and female students (aged 18-25) who visit the campus clinic at a university in KwaZulu-Natal. Both a descriptive survey and face to face interviews were used for data collection. In this study, it is affirmed that gender stratification, societal attitudes, and misconceptions about contraceptive use play an important role in the attitudes of young adults, male and female towards contraception and its use. Evidence of this is the high use of condoms amongst both male and female students' compared with other available methods. Among female students this was highly attributed to personal convenience and comfort with condom use as an unmarried young woman. It was clear from the data collected that respondents themselves attached some stigma to being associated with the use of contraceptive pills or having to visit the clinic regularly for injections as young unmarried women. Male respondents affirmed the use of the condom, although this was hardly with the view of taking reproductive/contraceptive responsibility, but rather, it was attributed to the function of the condom as a safe sex method that offered protection against sexually transmitted diseases and infections. Also evident from the study was the fact that male respondents felt more comfortable with their sexual functioning than the female respondents. This was easily attributed to the role of societal gender stratification in an individual's life. PMID- 17131605 TI - Western health practitioners' view about African traditional health practitioners' treatment and care of people living with HIV/AIDS. AB - African traditional health practitioners are an important source of health care for many South Africans. Thus, they are a health resource in this society. However, the integration of traditional health practitioners into the mainstream of health care is a complex process. Various factors contribute to this complexity, including the skepticism and reservation with which some western health practitioners view traditional health practitioners. This paper highlights the perceived strengths and weaknesses of the traditional healing system for people living with HIV/AIDS, as perceived by western health practitioners. The use of traditional practitioners as a choice of health care is attributed to both the strengths and weaknesses of this system of health care. The strength of the traditional healing system is in its sharing of the worldview and belief system of its users, it being an alternative to an inefficient western health care system (official system), privacy and absence of time limitations per consultation, treating patients psychologically, and scientifically unexplained physiological relief of the symptoms of specific illnesses. The perceived weaknesses of the traditional healing system include harmful treatment regimens, especially for people living with HIV/AIDS; prolonging the seeking of appropriate health care when traditional remedies fail to produce the desired effect; destroying interpersonal relationships of people living with HIV/AIDS through witchcraft accusations; psychological torment caused by the belief that HIV/AIDS can be cured by traditional remedies/intervention; and increasing the workload of western practitioners who are requested by patients to conduct multiple HIV tests after undergoing various traditional treatment regimens to cure HIV/AIDS. It is recommended that traditional practitioners be encouraged to adapt harmful traditional healing practices to the benefit of their patients in a non judgemental and non-critical manner. In addition, joint workshops should be conducted with traditional and western practitioners to demystify traditional healing practices. PMID- 17131606 TI - The leadership characteristics of the preceptor in selected clinical practice settings in Botswana. AB - A non-experimental, explorative, descriptive, quantitative study was undertaken. The purpose was to explore and describe the views of preceptors and preceptees regarding the fulfilment of the role of the preceptor in selected clinical nursing practice settings in the Botswana context. The study included 72 preceptors and 200 nursing students/preceptees who voluntary agreed voluntarily to participate in the study. A questionnaire was used to collect data, which was analyzed by using descriptive and inferential statistics. The findings of this study indicated that the preceptor lacked leadership characteristics in the accompaniment of the preceptee. These constraints included the lack of desirable characteristics such as intellectual, emotional, physical and other traits that are common to all good leaders. Recommendations were stated for improvements in selecting preceptors with certain leadership skills for the clinical practice settings. The limitations of this study were highlighted. PMID- 17131607 TI - Primigravidae's knowledge about obstetric complications in an urban health centre in Malawi. AB - Pregnant women in Malawi receive information about pregnancy, labour and delivery during routine antenatal visits. This study aimed to explore knowledge of obstetric complications amongst primigravidae attending an urban health centre in Blantyre, Malawi. A descriptive study design was used. Recognition of obstetric complications in pregnancy, during labour and after delivery and actions that participants would take if they developed any complications in pregnancy and after delivery were explored. Actions that women would take for complications that occur during labour were not probed, as women have little control over actions taken when complications arise during labour. METHODS: Participants were selected by means of purposive sampling from a population of pregnant women who fitted defined criteria and who were attending antenatal clinic at a health centre. Forty-five primigravidae from the urban setting with a gestation period between 28 and 42 weeks were interviewed. Data were analysed manually. RESULTS: The findings showed that participants were more aware of obstetric complications that could occur in pregnancy than of complications that may occur during and after delivery. Sixty percent of the participants were knowledgeable about obstetric complications in pregnancy. The majority of the participants, 73% and 82.2% did not know of any problems that could occur during and after the birth of the baby respectively. Participants had limited knowledge of complications that may need immediate treatment during all three periods. Fifty-eight percent (95% ci: 43; 73) of the primigravidae had some knowledge and could make an informed decision to go to a health facility with pregnancy complications. However, only 24% (95% ci: 11; 38) of the primigravidae had some knowledge and could make an informed decision to go to a health facility with complications after delivery. These findings suggest a critical need for provision of information on obstetric complications especially those that may occur during and after birth with emphasis on those obstetric complications that require immediate treatment. PMID- 17131608 TI - Differences regarding job satisfaction and job involvement of psychologists with different dominant career anchors. AB - In order to contribute to higher levels of job satisfaction, job involvement and productivity, a match or fit should be established between the dominant career anchor associated with a specific occupation and that of the employee. A career anchor is an individual's set of self-perceived talents, abilities, motives, needs and values that form the nucleus of one's occupational self-concept. Psychologists have always been part of the service orientated careers and therefore one would expect that it is likely that their dominant career anchor would be service orientation. If this is the case, psychologists with service as their dominant career anchor are supposed to have greater job satisfaction and job involvement compared to those with different career anchors. However, according to literature, this assumption is not necessarily correct. The primary goals of the current study were to determine whether in fact service is the dominant career anchor of psychologists in the Free State and whether there are significant differences regarding job satisfaction and job involvement between psychologists with and without service as their dominant career anchor. A third goal was to determine whether psychologists with different dominant career anchors differ significantly from one another regarding job satisfaction and job involvement. Questionnaires measuring career orientations, job satisfaction and job involvement were sent to 165 of the 171 registered psychologists in the Free State region. Only 75 psychologists (45,5%) responded which exceeded the traditional return rate of 20 to 30%. Due to the small sample of respondents, a nonparametric statistical test, namely the Mann Whitney U test was conducted to determine possible differences. An analysis of the data showed that 21 respondents had entrepreneurship as their dominant career orientation while 12 fell in the technical/functional, 12 in the challenging, 9 in the service and 8 in the autonomy categories of dominant career anchors. No significant differences regarding job satisfaction between psychologists with and without service as dominant career orientation could be determined. Both groups experienced a fairly high degree of job satisfaction and a higher level of intrinsic job satisfaction occurred compared to extrinsic job satisfaction. A significant difference between the two groups in terms of job involvement occurred. Psychologists with service as dominant career orientation showed a higher level of job involvement, although the degree of job involvement for both groups was fairly low. No significant differences regarding job satisfaction and job involvement among psychologists with different career orientations could be found. PMID- 17131609 TI - Exploration of the views of traditional healers regarding the termination of pregnancy (TOP) law. AB - The North-West Province is predominantly a rural area, and traditional healers remain the most important and influential members of the rural communities. A qualitative, explorative, descriptive and contextual research design was used. In depth, individual focused and interactive interviews were held with eight traditional healers from the rural areas of Mmabatho-Mafikeng. In addition, field notes and observations were utilised. The objective of this article is to explore the views of the traditional healers regarding termination of pregnancy (TOP) law. The results reflected the following themes: termination of pregnancy is killing; a child is a precious gift from God and the ancestors; there are alternatives to TOP; people who had any type of abortion should be cleansed with "dipitsa" or herbs; TOP may be allowed only in case of rape and incest, rape and incest offenders should be severely punished; and the traditional healers were not consulted during formulation of the TOP Law. It is therefore recommended that traditional should be involved in TOP workshops and educational programmes to enable them to provide counselling before and after abortion. PMID- 17131610 TI - Facilitation as a teaching strategy : experiences of facilitators. AB - Changes in nursing education involve the move from traditional teaching approaches that are teacher-centred to facilitation, a student centred approach. The student-centred approach is based on a philosophy of teaching and learning that puts the learner on centre-stage. The aim of this study was to identify the challenges of facilitators of learning using facilitation as a teaching method and recommend strategies for their (facilitators) development and support. A qualitative, explorative and contextual design was used. Four (4) universities in South Africa which utilize facilitation as a teaching/ learning process were identified and the facilitators were selected to be the sample of the study. The main question posed during in-depth group interviews was: How do you experience facilitation as a teaching/learning method?. Facilitators indicated different experiences and emotions when they first had to facilitate learning. All of them indicated that it was difficult to facilitate at the beginning as they were trained to lecture and that no format for facilitation was available. They experienced frustrations and anxieties as a result. The lack of knowledge of facilitation instilled fear in them. However they indicated that facilitation had many benefits for them and for the students. Amongst the ones mentioned were personal and professional growth. Challenges mentioned were the fear that they waste time and that they do not cover the content. It is therefore important that facilitation be included in the training of nurse educators. PMID- 17131611 TI - Knowledge of breast cancer in women in Sierra Leone. AB - Breast cancer has been described as one of the life-threatening diseases affecting women and is a major problem in women's health issues. The unrecorded number of cases of breast lumps and breast cancer observed in women in Sierra Leone prompted the researcher to organize a "Breast Week" during which 1,200 women were educated on breast cancer and the importance of breast health. This research is a follow up of the "Breast Week" which was organized in Freetown, Sierra Leone The specific objective of this study was to assess whether the knowledge and teachings given to the women who participated in this project was fully understood. A sample size of 120 women (10%) who participated in the "Breast Week" was obtained through systematic sampling. A quantitative approach was adopted and a structured interview schedule guided the data collection process. The data were processed through use of SPSS and Microsoft Excel. Texts from open ended questions were categorized and frequency counts were applied to the data. It was found that the majority (96.6%) of the women had some knowledge of breast cancer. They linked breast cancer to the signs and symptoms associated with it and were able to describe the disease as one that kills women if not promptly detected and/or treated appropriately. Findings indicate that the majority of the women are aware of the dangers of the disease and had knowledge of someone who had died of breast cancer (59.2%). An assessment of the effectiveness of knowledge on breast cancer showed that these women could identify breast cancer as a disease that affects women and may cause death if not detected on time. PMID- 17131612 TI - Higher order thinking skills competencies required by outcomes-based education from learners. AB - Outcomes-Based Education (OBE) brought about a significant paradigm shift in the education and training of learners in South Africa. OBE requires a shift from focusing on the teacher input (instruction offerings or syllabuses expressed in terms of content), to focusing on learner outcomes. OBE is moving away from 'transmission' models to constructivistic, learner-centered models that put emphasis on learning as an active process (Nieburh, 1996:30). Teachers act as facilitators and mediators of learning (Norms and Standards, Government Gazette vol 415, no 20844 of 2000). Facilitators are responsible to create the environment that is conducive for learners to construct their own knowledge, skills and values through interaction (Peters, 2000). The first critical cross field outcome accepted by the South African Qualification Framework (SAQA) is that learners should be able to identify and solve problems by using critical and creative thinking skills. This paper seeks to explore some higher order thinking skills competencies required by OBE from learners such as critical thinking, reflective thinking, creative thinking, dialogic / dialectic thinking, decision making, problem solving and emotional intelligence and their implications in facilitating teaching and learning from the theoretical perspective. The philosophical underpinning of these higher order thinking skills is described to give direction to the study. It is recommended that a study focusing on the assessment of these intellectual concepts be made. The study may be qualitative, quantitative or mixed methods in nature (Creswell 2005). PMID- 17131613 TI - The views of undergraduate nursing students on caring for patients with HIV/AIDS. AB - A qualitative, exploratory and descriptive study was conducted with the purpose of exploring and describing the views of undergraduate nursing students on caring for HIV/AIDS patients. The study population consisted of BCur III nursing students studying at the Medical University of Southern Africa (Medunsa). Participants were purposively selected. Focus group interviews were used as a data collection instrument. Guided by a group moderator and responding to a central research question, participants shared their views about caring for HIV/AIDS patients. Tesch's qualitative method of data analysis, as described by Cresswell (1994:155), was used to analyse the data. Caring for HIV/AIDS patients evoked emotions such as fear, anger and frustration among undergraduate nursing students. Students expressed needs such as the acquisition of knowledge and a reduction in the stigmatisation of patients with HIV/ AIDS, while the data analysis revealed demands such as more intensive clinical accompaniment by lecturers and antiretroviral therapy delivery by government. Suggested solutions included student participation in HIV/AIDS prevention campaigns and the upholding of patients' rights. Curriculum innovation was recommended to improve students' knowledge of HIV/AIDS and to ensure the provision of quality care for these patients. PMID- 17131614 TI - Does a 'shadow workforce' of inactive nurses exist? AB - The entire population of inactive nurses in Vermont was surveyed to determine if a "shadow workforce" exists. The notion that large numbers of nurses are available to return to work is not supported by this study. Desirable benefits for those wishing to return are discussed. PMID- 17131615 TI - Hospital nursing costs, billing, and reimbursement. AB - Nursing intensity, estimated direct nursing costs, and daily billing were compared for 12 adult medical or surgical units at an academic medical center from January 1 to May 31, 2005 (22,649 patient days). Two main findings, nursing intensity and direct nursing costs, were highly variable within and across each of the study nursing units (mean 429 dollars, SD 160 dollars); direct costs of nursing care were significantly higher for private room rates compared to intermediate room per diem charges billed at a higher rate (441 dollars vs. 426 dollars, F 37.77, p < 0.001). The results demonstrate that the direct costs of nursing care are not aligned with current billing practices at this university hospital. The use of fixed room and board charges to account for nursing care in U.S. hospitals may be obsolete and an alternative nurse-centric costing, billing, and reimbursement model is proposed. PMID- 17131616 TI - Influence of hospital context on cost and length of stay following cardiac catheterization. AB - This study attempted to link health care activities and outcomes, taking into account characteristics of the health care system, the work group, and the individual. Results indicated that organizational structure and work group process had a greater influence on cost and derived length of stay (a measure of efficiency) than did the individual-level variables. The findings of this study add to the body of knowledge that explains how the processes of nurses' work influence the outcomes of that work. PMID- 17131617 TI - A dramaturgical analysis of shift report patterns with cost implications: a case study. AB - This secondary analysis uses Goffman's stage drama to explore the roles of registered nurses in videotaped reports in a hospital. Each report was a performance by the charge nurse which affected the climate, interactions, and duration of the report. PMID- 17131618 TI - Evaluating recruitment process through 'Mystery Shops'. PMID- 17131619 TI - High-performance health care: how do we get there? PMID- 17131620 TI - Improving clinical communication through structured conversation. AB - Communication failure plays an important role in causing medical errors. Clinical communication is highly complex and prone to error especially during transitions of patient care and emergent situations. Standardized approaches and tools may provide potential solutions to improve the quality of communication and prevent subsequent patient harm. The National Patient Safety Foundation, through its Stand Up for Patient Safety Program, is committed to assisting organizations improve clinical communication. PMID- 17131621 TI - Innovation lies awaiting. PMID- 17131622 TI - Reliability between nurse managers: the key to the high-reliability organization. AB - Flawless execution rests in the hands of nurse managers. No one can work alone in health care any more. We are interdependent and know that the best outcomes happen when practices are organized around collegial supportive structures rather than autonomous competitive units. We are only as strong as our weakest link. If all managers see the big picture and look beyond their units for what is right for the common good, we will achieve high-reliability organizations in health care. In turn health care organizations will become very safe places to operate. Shared governance structures for nurse managers are the perfect vehicle to develop collaborative organizations and flawless execution, and to adopt high reliability organization principles. PMID- 17131623 TI - Tumor markers in detection of lung cancer. AB - The expression of biomarkers by lung cancers is useful in the diagnosis and clinical management of patients with lung cancer. Biomarkers provide insight into histogenesis, interrelationships, and biological behavior of lung tumors. This chapter presents data on lung cancer detection, involving some of the most studied and interesting lung cancer biomarkers to date-CYFRA 21-1, NSE, ProGRP, SCC, CEA, Tumor M2-PK, as well as markers in clinical application such as CRP, LDH, tumor-suppressor genes and oncogenes, CA125, CgA, NCAM, and TPA. Biomarker profiles in combination with fuzzy logic techniques have also been addressed. Serum markers used alone or in combination with other indices might play an important role in monitoring response to therapy in early detection of tumor reactivation in new treatment strategies as well as in secondary prevention. PMID- 17131624 TI - Advances in serum protein electrophoresis. AB - A major advance in serum protein electrophoresis in the last decade has been the introduction of capillary zone electrophoresis (CZE). Two dedicated automated multichannel instruments for serum protein separation by CZE in clinical laboratories are available, the Paragon CZE 2000 (Beckman Coulter, CA) and the Capillarys (Sebia, France). This chapter focuses on the performance of these commercial multichannel CZE systems. Following topics are addressed: precision, comparison of CZE with gel-based methods, dysproteinemia analysis by CZE, detection and identification of monoclonal proteins by CZE, quality control, and interferences. Examples of electrophoretic patterns are given. PMID- 17131625 TI - Aspirin resistance: a review of diagnostic methodology, mechanisms, and clinical utility. AB - Ingestion of a daily aspirin in patients with coronary artery disease decreases the rate of occlusive atherosclerotic events by about 25 percent. Some patients whose platelets are minimally inhibited by aspirin are categorized as aspirin resistant. Three reports document an increased risk for future vascular events in aspirin resistant patients. Aspirin's platelet inhibitory effect is measured using a variety of techniques. The demarcation between minimal and expected aspirin inhibition of platelets is arbitrarily determined by each investigator which leads to confusion in translating these reports to patient care. The focus of this report is the relative merits of the different techniques and their utility for defining patients with minimal aspirin induced platelet inhibition. The clinically useful mechanisms underlying decreased aspirin induced platelet inhibition include failure of a patient to take their daily aspirin, poor compliance, and nonsteroidal anti-inflammatory drugs (NSAIDs) interference with aspirin's ability to get to its binding site on the cyclooxygenase enzyme-1 (COX 1)]. Compliance is best assessed by comparing the results obtained with arachidonic acid (AA) stimulated light aggregation at two time points. The first time point is while the patient is supposedly taking their usual daily aspirin and the second time point is 2 hours after the patient is observed to ingest 325 mg of aspirin. After observed ingestion of aspirin, those patients with minimal aspirin inhibition of platelets are best detected using light aggregation stimulated by a new platelet agonist platelet prostaglandin agonist (PPA). In order for the results of a particular technique to be clinically meaningful it must be shown that those patients with minimal aspirin inhibition of platelets have an increased risk for a future vascular event that is independent from known major cardiovascular risk factors. PMID- 17131626 TI - Vitreous fluid biomarkers. AB - Interactions between cells and the network of secreted proteins are associated with the ocular disease. In most cases, clinical appearcance is sufficiently diagnostic. However, in cases of nonspecific or atypical clinical presentation, diagnostic sampling of vitreous fluid can aid diagnosis and treatment for ocular disease. Progresses in the basic sciences, particularly molecular biology, and advances in surgical instrumentation have greatly enhanced the diagnostic armamentarium. These developments also have led to a better understanding of the pathophysiological processes involved in ocular diseases and have prompted evolution of new therapeutic modalities. In this chapter, we review techniques for vitreous fluid sampling and biomarker quantitation thereof. The molecular biology of bioactive vitreous fluid factors is also discussed with respect to their clinical involvement in the development of ocular disease. PMID- 17131627 TI - Tumor-associated carbonic anhydrases and their clinical significance. AB - Carbonic anhydrases (CAs) are physiologically important enzymes that catalyze a reversible conversion of carbon dioxide to bicarbonate and participate in ion transport and pH control. Two human isoenzymes, CA IX and CA XII, are overexpressed in cancer and contribute to tumor physiology. Particularly CA IX is confined to only few normal tissues but is ectopically induced in many tumor types mainly due to its strong transcriptional activation by hypoxia accomplished via HIF-1 transcription factor. Therefore, CA IX can serve as a surrogate marker of hypoxia and a prognostic indicator. CA IX appears implicated in cell adhesion and in balance of pH disturbances caused by tumor metabolism. Both tumor-related expression pattern and functional involvement in tumor progression make it a suitable target for anticancer treatment. Here we summarize a current knowledge on CA IX and CA XII, and discuss possibilities of their exploitation for cancer detection, diagnostics, and therapy. PMID- 17131628 TI - Mitochondrial disease: maintenance of mitochondrial genome and molecular diagnostics. AB - Mitochondrial DNA (mtDNA) is essential for the aerobic ATP synthesis system that is responsible for about 80% of normal cellular energy demands. In addition to rare genetic disorders causing neuromyopathy, alterations of mtDNA have been found also in so-called common diseases such as heart failure, diabetes, and cancer. Although some of these alterations are inherited, some are considered to be generated and/or accumulated in somatic cells with age. One reason for the somatic mutations is that mtDNA is more vulnerable than is nuclear DNA. For example, mitochondrial respiratory chain produces a large amount of reactive oxygen species as inevitable byproducts of oxidative phosphorylation. However, the molecular mechanisms for maintenance of mitochondrial genome are much less elucidated than those for nuclear genome. In spite of its increasing importance, the molecular diagnosis of mitochondrial DNA-related diseases is well done only in very limited expert laboratories. In this chapter, we focus on maintenance of mtDNA in somatic cells, its clinical importance, and recent developments of molecular tests. PMID- 17131629 TI - Microfluidic enzyme-linked immunosorbent assay technology. AB - In this chapter, we have presented an overview of microfluidic enzyme-linked immunosorbent assay (ELISA) by first introducing the principle of immunoassay, ELISA, and microfabricated devices, followed by a discussion of microfabrication technology and the characterization of microfluidic components. Significant advances in laboratory technology are contributing to the further understanding of microfluidic function, surface modification and immobilization, which lead to the development of improved biomolecule detection methods and prospective applications. For the future, the exploitation of more robust-manufacturing processes and integrated assay systems in an automatic fashion with much reduced assay time and reagent consumption will allow for the effective detection and quantification of biological agents that are of interest in medical diagnostics, food safety surveillance, and environmental monitoring. PMID- 17131630 TI - Pathogenic mechanisms of anti-endothelial cell antibodies (AECA): their prevalence and clinical relevance. AB - Anti-endothelial cell antibodies (AECA) represent a heterogeneous family of autoantibodies directed against structural endothelial proteins, as well as antigens adhering to endothelial cells. Although AECA immunoassays still show a high-interlaboratory variability, several findings suggest a pathogenic role of these autoantibodies in diseases characterized by endothelial damage. In this chapter, we analyze the knowledge about AECA prevalence, clinical relevance, and their pathogenic role in autoimmune diseases focusing in particular on systemic lupus erythematosus, antiphospholipid syndrome, systemic sclerosis (SSc), and systemic vasculitis. PMID- 17131631 TI - Prioritising postnatal care. PMID- 17131632 TI - Don't demonise abuse survivors who really need caesareans consider the amazing possibilities for you. PMID- 17131634 TI - Celebrations as Stroud Maternity Unit is saved. PMID- 17131633 TI - Health Minister criticises RCM. PMID- 17131635 TI - Infant feeding trends: early indicators. PMID- 17131636 TI - Best buddies during birth and breastfeeding. PMID- 17131637 TI - La Leche League: the ultimate mother's help. PMID- 17131638 TI - Nurturing new mothers as well as babies. PMID- 17131639 TI - Vaginal birth after caesarean (part 2). PMID- 17131640 TI - Diet in pregnancy, labour and beyond. PMID- 17131641 TI - A healthy preconception. PMID- 17131642 TI - Midwifery basics: women's health needs (2). Young women. PMID- 17131643 TI - Music for pain relief. PMID- 17131644 TI - Holistic therapies, proof and plausibility. PMID- 17131645 TI - Who and what is a midwife? PMID- 17131646 TI - Detection of irregular spatial structures. AB - Wilson et al.'s (1997) study on Glass patterns suggested that the integration of stimulus features into a linear shape occurs quite locally, whereas curved structures--such as circular--require global summation. Their conclusion was based on experiments in which they varied the size of the signal area containing a spatial structure. In the present study, we tested the integration of constant sized linear and curved Glass patterns by varying their global irregularity. If the mechanisms underlying the detection of a Glass pattern pool features globally throughout the stimulus, the irregularity should have a strong effect on detection performance. The irregular Glass patterns were composed of a variable number of sub-areas, each of which contained its own linear or curved structure. The structural irregularity impaired the detection of the curved patterns, whereas the thresholds for the linear patterns were not affected. Thus, our results are in line with the notion that the integration of curved Glass patterns occurs more globally than the integration of linear patterns. PMID- 17131647 TI - Texture discrimination asymmetries across the visual field. AB - Texture discrimination is sometimes asymmetrical; texture A embedded in texture B is more easily detected than texture B embedded in texture A. Furthermore, texture discrimination often improves as the disparate texture is moved into the periphery; this has been referred to as the central performance drop (CPD). The interaction of these interesting and counter-intuitive aspects of texture discrimination has received very little attention. Using four stimulus pattern pairs that were previously shown to elicit asymmetrical texture discrimination, we examined texture discrimination asymmetries as a function of eccentricity. We found three patterns of results; (i) both texture arrangements (A in B, and B in A) elicit a CPD but do not show an asymmetry, (ii) both texture arrangements elicit a monotonic decrease in performance with eccentricity (i.e. no CPD) but an asymmetry is seen at each eccentricity and (iii) discrimination asymmetries are minimal at fixation and in the far periphery and maximal about 3 degrees from fixation with a CPD generally shown for the 'stronger' member of the pair. These results emphasize that one cannot talk about the 'discriminability' of a particular texture pair without reference to the arrangement of the two textures and the eccentricity of presentation. PMID- 17131648 TI - Psychometric curves of lateral facilitation. AB - Visibility of an oriented stimulus may be enhanced by nearby stimuli that are co aligned with the target. However, the underlying mechanism governing this facilitation is controversial. Here we measured the dependence of percent correct detection on the target's contrast (psychometric curve) with and without flankers, where flankers were either collinear with or orthogonal to the target. We find that the effect of collinear flankers can be described as a translation of the psychometric function along the linear contrast axis. This behavior is consistent, within experimental error, with two types of models: (1) non-linear transduction of target contrast with collinear flankers having additive effects on contrast, and (2) uncertainty reduction (Pelli, 1985) by collinear flankers. We discuss properties of collinear facilitation that can help deciding between these two models. PMID- 17131649 TI - A 'first stage' central performance drop in a Gabor luminance-modulation detection task. AB - In an experiment, 20 participants had to detect a backward masked Gabor luminance modulation target imposed on a field of uniform luminance at varying eccentricities along the horizontal meridian. Different spatial frequencies were used as target modulations. Results for a 7.0 c/deg target patch showed peak detection performance at the center of the visual field and a steady decrease toward the periphery. For 1.0 c/deg, 0.75 c/deg, and 0.5 c/deg target patches, in contrast, the peak was several degrees off retinal center and decreased steadily toward the center. Findings not only confirmed the familiar sensitivity loss toward peripheral areas for high spatial frequencies, but also indicated a sensitivity loss toward central areas for low spatial frequencies. It is concluded that they further support Gurnsey et al.'s (1996) 'mismatch hypothesis' extending its scope to also include 'first-stage' stimuli. PMID- 17131650 TI - Perceptual distance and the constancy of size and stereoscopic depth. AB - The relationship between distance and size perception is unclear because of conflicting results of tests investigating the size-distance invariance hypothesis (SDIH), according to which perceived size is proportional to perceived distance. We propose that response bias with regard to measures of perceived distance is at the root of the conflict. Rather than employ the usual method of magnitude estimation, the bias-free two-alternative forced choice (2AFC) method was used to determine the precision (1/sigma) of discriminating depth at different distances. The results led us to define perceptual distance as a bias free power function of physical distance, with an exponent of approximately 0.5. Similar measures involving size differences among stimuli of equal angular size yield the same power function of distance. In addition, size discrimination is noisier than depth discrimination, suggesting that distance information is processed prior to angular size. Size constancy implies that the perceived size is proportional to perceptual distance. Moreover, given a constant relative disparity, depth constancy implies that perceived depth is proportional to the square of perceptual distance. However, the function relating the uncertainties of depth and of size discrimination to distance is the same. Hence, depth and size constancy may be accounted for by the same underlying law. PMID- 17131651 TI - Cueing of object orientation facilitates attentional selection of relevant objects. AB - Three experiments investigated the effects of advance information about orientation on the processing of relevant and irrelevant objects, as indicated by Stroop effects from color words located in either object. Four results were obtained. First, participants showed the expected modulation of the Stroop effect: words in the relevant object produced much larger Stroop effects than words in the irrelevant object or words in the background. Second, blocking of object orientation had no effects. Third, informative orientation cues facilitated processing of the relevant object, but cueing did not affect processing of the irrelevant object. Fourth, effects of informative orientation cues were restricted to the first part of each experiment. Results suggest that observers can use advance information about object orientation for improving attentional selection of a visual object. In addition, the results revealed some constraints for the effective use of orientation cues, and discard possible explanations for the observed modulation of Stroop effects. PMID- 17131652 TI - Evaluation of neural network robust reliability using information-gap theory. AB - A novel technique for the evaluation of neural network robustness against uncertainty using a nonprobabilistic approach is presented. Conventional optimization techniques were employed to train multilayer perceptron (MLP) networks, which were then probed with an uncertainty analysis using an information-gap model to quantify the network response to uncertainty in the input data. It is demonstrated that the best performing network on data with low uncertainty is not in general the optimal network on data with a higher degree of input uncertainty. Using the concepts of information-gap theory, this paper develops a theoretical framework for information-gap uncertainty applied to neural networks, and explores the practical application of the procedure to three sample cases. The first consists of a simple two-dimensional (2-D) classification network operating on a known Gaussian distribution, the second a nine-lass vibration classification problem from an aircraft wing, and the third a two-class example from a database of breast cancer incidence. PMID- 17131653 TI - Global convergence of decomposition learning methods for support vector machines. AB - Decomposition methods are well-known techniques for solving quadratic programming (QP) problems arising in support vector machines (SVMs). In each iteration of a decomposition method, a small number of variables are selected and a QP problem with only the selected variables is solved. Since large matrix computations are not required, decomposition methods are applicable to large QP problems. In this paper, we will make a rigorous analysis of the global convergence of general decomposition methods for SVMs. We first introduce a relaxed version of the optimality condition for the QP problems and then prove that a decomposition method reaches a solution satisfying this relaxed optimality condition within a finite number of iterations under a very mild condition on how to select variables. PMID- 17131654 TI - The fastICA algorithm revisited: convergence analysis. AB - The fast independent component analysis (FastICA) algorithm is one of the most popular methods to solve problems in ICA and blind source separation. It has been shown experimentally that it outperforms most of the commonly used ICA algorithms in convergence speed. A rigorous local convergence analysis has been presented only for the so-called one-unit case, in which just one of the rows of the separating matrix is considered. However, in the FastICA algorithm, there is also an explicit normalization step, and it may be questioned whether the extra rotation caused by the normalization will affect the convergence speed. The purpose of this paper is to show that this is not the case and the good convergence properties of the one-unit case are also shared by the full algorithm with symmetrical normalization. A local convergence analysis is given for the general case, and the global behavior is illustrated numerically for two sources and two mixtures in several typical cases. PMID- 17131655 TI - Locally weighted interpolating growing neural gas. AB - In this paper, we propose a new approach to function approximation based on a growing neural gas (GNG), a self-organizing map (SOM) which is able to adapt to the local dimension of a possible high-dimensional input distribution. Local models are built interpolating between values associated with the map's neurons. These models are combined using a weighted sum to yield the final approximation value. The values, the positions, and the "local ranges" of the neurons are adapted to improve the approximation quality. The method is able to adapt to changing target functions and to follow nonstationary input distributions. The new approach is compared to the radial basis function (RBF) extension of the growing neural gas and to locally weighted projection regression (LWPR), a state of-the-art algorithm for incremental nonlinear function approximation. PMID- 17131656 TI - FCMAC-Yager: a novel Yager-inference-scheme-based fuzzy CMAC. AB - The cerebellum is a brain region important for a number of motor and cognitive functions. It is able to generate error correction signals to drive learning and for the acquisition of memory skills. The cerebellar model articulation controller (CMAC) is a neural network inspired by the neurophysiologic theory of the cerebellum and is recognized for its localized generalization and rapid algorithmic computation capabilities. The main deficiencies in the basic CMAC structure are: (1) it is difficult to interpret the internal operations of the CMAC network and (2) the resolution (quantization) problem arising from the partitioning of the input training space. These limitations lead to the synthesis of a fuzzy quantization technique and the mapping of a fuzzy inference scheme onto the CMAC structure. The discrete incremental clustering (DIC) technique is employed to alleviate the quantization problem in the CMAC structure, resulting in the fuzzy CMAC (FCMAC) network. The Yager inference scheme (Yager), which possesses firm fuzzy logic foundation and maps closely to the logical implication operations in the classical (binary) logic framework, is subsequently mapped onto the FCMAC structure. This results in a novel fuzzy neural architecture known as thefuzzy cerebellar model articulation controller-Yager (FCMAC-Yager) system. The proposed FCMAC-Yager network exhibits learning and memory capabilities of the cerebellum through the CMAC structure while emulating the human way of reasoning through the Yager. The new FCMAC-Yager network employs a two-phase training algorithm consisting of structural learning based on the DIC technique and parameter learning using hebbian learning (associative long-term potentiation). The proposed FCMAC-Yager architecture is evaluated using an extensive suite of real-life applications such as highway traffic-trend modeling and prediction and performing as an early warning system for bank failure classification and medical diagnosis of breast cancer. The experimental results are encouraging. PMID- 17131657 TI - A fast and accurate online sequential learning algorithm for feedforward networks. AB - In this paper, we develop an online sequential learning algorithm for single hidden layer feedforward networks (SLFNs) with additive or radial basis function (RBF) hidden nodes in a unified framework. The algorithm is referred to as online sequential extreme learning machine (OS-ELM) and can learn data one-by-one or chunk-by-chunk (a block of data) with fixed or varying chunk size. The activation functions for additive nodes in OS-ELM can be any bounded nonconstant piecewise continuous functions and the activation functions for RBF nodes can be any integrable piecewise continuous functions. In OS-ELM, the parameters of hidden nodes (the input weights and biases of additive nodes or the centers and impact factors of RBF nodes) are randomly selected and the output weights are analytically determined based on the sequentially arriving data. The algorithm uses the ideas of ELM of Huang et al. developed for batch learning which has been shown to be extremely fast with generalization performance better than other batch training methods. Apart from selecting the number of hidden nodes, no other control parameters have to be manually chosen. Detailed performance comparison of OS-ELM is done with other popular sequential learning algorithms on benchmark problems drawn from the regression, classification and time series prediction areas. The results show that the OS-ELM is faster than the other sequential algorithms and produces better generalization performance. PMID- 17131658 TI - Parameter incremental learning algorithm for neural networks. AB - In this paper, a novel stochastic (or online) training algorithm for neural networks, named parameter incremental learning (PIL) algorithm, is proposed and developed. The main idea of the PIL strategy is that the learning algorithm should not only adapt to the newly presented input-output training pattern by adjusting parameters, but also preserve the prior results. A general PIL algorithm for feedforward neural networks is accordingly presented as the first order approximate solution to an optimization problem, where the performance index is the combination of proper measures of preservation and adaptation. The PIL algorithms for the multilayer perceptron (MLP) are subsequently derived. Numerical studies show that for all the three benchmark problems used in this paper the PIL algorithm for MLP is measurably superior to the standard online backpropagation (BP) algorithm and the stochastic diagonal Levenberg-Marquardt (SDLM) algorithm in terms of the convergence speed and accuracy. Other appealing features of the PIL algorithm are that it is computationally as simple as the BP algorithm, and as easy to use as the BP algorithm. It, therefore, can be applied, with better performance, to any situations where the standard online BP algorithm is applicable. PMID- 17131659 TI - A hybrid forward algorithm for RBF neural network construction. AB - This paper proposes a novel hybrid forward algorithm (HFA) for the construction of radial basis function (RBF) neural networks with tunable nodes. The main objective is to efficiently and effectively produce a parsimonious RBF neural network that generalizes well. In this study, it is achieved through simultaneous network structure determination and parameter optimization on the continuous parameter space. This is a mixed integer hard problem and the proposed HFA tackles this problem using an integrated analytic framework, leading to significantly improved network performance and reduced memory usage for the network construction. The computational complexity analysis confirms the efficiency of the proposed algorithm, and the simulation results demonstrate its effectiveness. PMID- 17131660 TI - An optimization methodology for neural network weights and architectures. AB - This paper introduces a methodology for neural network global optimization. The aim is the simultaneous optimization of multilayer perceptron (MLP) network weights and architectures, in order to generate topologies with few connections and high classification performance for any data sets. The approach combines the advantages of simulated annealing, tabu search and the backpropagation training algorithm in order to generate an automatic process for producing networks with high classification performance and low complexity. Experimental results obtained with four classification problems and one prediction problem has shown to be better than those obtained by the most commonly used optimization techniques. PMID- 17131661 TI - Associative learning in hierarchical self-organizing learning arrays. AB - In this paper, we introduce feedback-based associative learning in self-organized learning arrays (SOLAR). SOLAR structures are hierarchically organized networks of sparsely connected neurons that define their own functions and select their interconnections locally. This paper provides a description of neuron self organization and signal processing. Feedforward processing is used to make necessary correlations and learn the input patterns. Discovered associations between neuron inputs are used to generate feedback signals. These feedback signals, when propagated to the primary inputs, can establish the expected input values. This can be used for heteroassociative (HA) and autoassociative (AA) learning and pattern recognition. Example applications in HA learning are given. PMID- 17131662 TI - Convergence of neural networks for programming problems via a nonsmooth Lojasiewicz inequality. AB - This paper considers a class of neural networks (NNs) for solving linear programming (LP) problems, convex quadratic programming (QP) problems, and nonconvex QP problems where an indefinite quadratic objective function is subject to a set of affine constraints. The NNs are characterized by constraint neurons modeled by ideal diodes with vertical segments in their characteristic, which enable to implement an exact penalty method. A new method is exploited to address convergence of trajectories, which is based on a nonsmooth Lojasiewicz inequality for the generalized gradient vector field describing the NN dynamics. The method permits to prove that each forward trajectory of the NN has finite length, and as a consequence it converges toward a singleton. Furthermore, by means of a quantitative evaluation of the Lojasiewicz exponent at the equilibrium points, the following results on convergence rate of trajectories are established: (1) for nonconvex QP problems, each trajectory is either exponentially convergent, or convergent in finite time, toward a singleton belonging to the set of constrained critical points; (2) for convex QP problems, the same result as in (1) holds; moreover, the singleton belongs to the set of global minimizers; and (3) for LP problems, each trajectory converges in finite time to a singleton belonging to the set of global minimizers. These results, which improve previous results obtained via the Lyapunov approach, are true independently of the nature of the set of equilibrium points, and in particular they hold even when the NN possesses infinitely many nonisolated equilibrium points. PMID- 17131663 TI - Solving pseudomonotone variational inequalities and pseudoconvex optimization problems using the projection neural network. AB - In recent years, a recurrent neural network called projection neural network was proposed for solving monotone variational inequalities and related convex optimization problems. In this paper, we show that the projection neural network can also be used to solve pseudomonotone variational inequalities and related pseudoconvex optimization problems. Under various pseudomonotonicity conditions and other conditions, the projection neural network is proved to be stable in the sense of Lyapunov and globally convergent, globally asymptotically stable, and globally exponentially stable. Since monotonicity is a special case of pseudomononicity, the projection neural network can be applied to solve a broader class of constrained optimization problems related to variational inequalities. Moreover, a new concept, called componentwise pseudomononicity, different from pseudomononicity in general, is introduced. Under this new concept, two stability results of the projection neural network for solving variational inequalities are also obtained. Finally, numerical examples show the effectiveness and performance of the projection neural network. PMID- 17131664 TI - A simplified dual neural network for quadratic programming with its KWTA application. AB - The design, analysis, and application of a new recurrent neural network for quadratic programming, called simplified dual neural network, are discussed. The analysis mainly concentrates on the convergence property and the computational complexity of the neural network. The simplified dual neural network is shown to be globally convergent to the exact optimal solution. The complexity of the neural network architecture is reduced with the number of neurons equal to the number of inequality constraints. Its application to k-winners-take-all (KWTA) operation is discussed to demonstrate how to solve problems with this neural network. PMID- 17131665 TI - Neural networks for continuous online learning and control. AB - This paper proposes a new hybrid neural network (NN) model that employs a multistage online learning process to solve the distributed control problem with an infinite horizon. Various techniques such as reinforcement learning and evolutionary algorithm are used to design the multistage online learning process. For this paper, the infinite horizon distributed control problem is implemented in the form of real-time distributed traffic signal control for intersections in a large-scale traffic network. The hybrid neural network model is used to design each of the local traffic signal controllers at the respective intersections. As the state of the traffic network changes due to random fluctuation of traffic volumes, the NN-based local controllers will need to adapt to the changing dynamics in order to provide effective traffic signal control and to prevent the traffic network from becoming overcongested. Such a problem is especially challenging if the local controllers are used for an infinite horizon problem where online learning has to take place continuously once the controllers are implemented into the traffic network. A comprehensive simulation model of a section of the Central Business District (CBD) of Singapore has been developed using PARAMICS microscopic simulation program. As the complexity of the simulation increases, results show that the hybrid NN model provides significant improvement in traffic conditions when evaluated against an existing traffic signal control algorithm as well as a new, continuously updated simultaneous perturbation stochastic approximation-based neural network (SPSA-NN). Using the hybrid NN model, the total mean delay of each vehicle has been reduced by 78% and the total mean stoppage time of each vehicle has been reduced by 84% compared to the existing traffic signal control algorithm. This shows the efficacy of the hybrid NN model in solving large-scale traffic signal control problem in a distributed manner. Also, it indicates the possibility of using the hybrid NN model for other applications that are similar in nature as the infinite horizon distributed control problem. PMID- 17131666 TI - Developmental learning with behavioral mode tuning by carrier-frequency modulation in coherent neural networks. AB - We propose a developmental learning architecture with which a motion-control system learns multiple tasks similar to each other or advanced ones incrementally and efficiently by tuning its behavioral mode. The system is based on a coherent neural network whose carrier frequency works as a mode-tuning parameter. In our experiments, we consider two tasks related to bicycle riding. The first is to ride as temporally long as the system can before it falls down (task 1). The second is an advanced one, i.e., to ride as far as possible in a certain direction (task 2). We compare developmental learning to learn task 2 after task 1 with the direct learning of task 2. We also examine the effect of the mode tuning by comparing variable-mode learning (VML), where the carrier frequency is set free to move, with fixed-mode learning (FML), where the frequency is unchanged. We find that VML developmental learning results in the most efficient learning among the possible combinations. We discuss the effects of the incremental task assignment as well as the behavioral mode tuning in developmental learning. PMID- 17131667 TI - One-class-at-a-time removal sequence planning method for multiclass classification problems. AB - Using dynamic programming, this work develops a one-class-at-a-time removal sequence planning method to decompose a multiclass classification problem into a series of two-class problems. Compared with previous decomposition methods, the approach has the following distinct features. First, under the one-class-at-a time framework, the approach guarantees the optimality of the decomposition. Second, for a K-class problem, the number of binary classifiers required by the method is only K-1. Third, to achieve higher classification accuracy, the approach can easily be adapted to form a committee machine. A drawback of the approach is that its computational burden increases rapidly with the number of classes. To resolve this difficulty, a partial decomposition technique is introduced that reduces the computational cost by generating a suboptimal solution. Experimental results demonstrate that the proposed approach consistently outperforms two conventional decomposition methods. PMID- 17131668 TI - Discriminative common vector method with kernels. AB - In some pattern recognition tasks, the dimension of the sample space is larger than the number of samples in the training set. This is known as the "small sample size problem". Linear discriminant analysis (LDA) techniques cannot be applied directly to the small sample size case. The small sample size problem is also encountered when kernel approaches are used for recognition. In this paper, we attempt to answer the question of "How should one choose the optimal projection vectors for feature extraction in the small sample size case?" Based on our findings, we propose a new method called the kernel discriminative common vector method. In this method, we first nonlinearly map the original input space to an implicit higher dimensional feature space, in which the data are hoped to be linearly separable. Then, the optimal projection vectors are computed in this transformed space. The proposed method yields an optimal solution for maximizing a modified Fisher's linear discriminant criterion, discussed in the paper. Thus, under certain conditions, a 100% recognition rate is guaranteed for the training set samples. Experiments on test data also show that, in many situations, the generalization performance of the proposed method compares favorably with other kernel approaches. PMID- 17131669 TI - Permutation coding technique for image recognition systems. AB - A feature extractor and neural classifier for image recognition systems are proposed. The proposed feature extractor is based on the concept of random local descriptors (RLDs). It is followed by the encoder that is based on the permutation coding technique that allows to take into account not only detected features but also the position of each feature on the image and to make the recognition process invariant to small displacements. The combination of RLDs and permutation coding permits us to obtain a sufficiently general description of the image to be recognized. The code generated by the encoder is used as an input data for the neural classifier. Different types of images were used to test the proposed image recognition system. It was tested in the handwritten digit recognition problem, the face recognition problem, and the microobject shape recognition problem. The results of testing are very promising. The error rate for the Modified National Institute of Standards and Technology (MNIST) database is 0.44% and for the Olivetti Research Laboratory (ORL) database it is 0.1%. PMID- 17131670 TI - A new adaptive backpropagation algorithm based on Lyapunov stability theory for neural networks. AB - A new adaptive backpropagation (BP) algorithm based on Lyapunov stability theory for neural networks is developed in this paper. It is shown that the candidate of a Lyapunov function V(k) of the tracking error between the output of a neural network and the desired reference signal is chosen first, and the weights of the neural network are then updated, from the output layer to the input layer, in the sense that deltaV(k) = V(k) - V(k - 1) < 0. The output tracking error can then asymptotically converge to zero according to Lyapunov stability theory. Unlike gradient-based BP training algorithms, the new Lyapunov adaptive BP algorithm in this paper is not used for searching the global minimum point along the cost function surface in the weight space, but it is aimed at constructing an energy surface with a single global minimum point through the adaptive adjustment of the weights as the time goes to infinity. Although a neural network may have bounded input disturbances, the effects of the disturbances can be eliminated, and asymptotic error convergence can be obtained. The new Lyapunov adaptive BP algorithm is then applied to the design of an adaptive filter in the simulation example to show the fast error convergence and strong robustness with respect to large bounded input disturbances. PMID- 17131671 TI - Multiuser receiver for DS-CDMA signals in multipath channels: an enhanced multisurface method. AB - This paper deals with the problem of multiuser detection in direct-sequence code division multiple-access (DS-CDMA) systems in multipath environments. The existing multiuser detectors can be divided into two categories: (1) low complexity poor-performance linear detectors and (2) high-complexity good performance nonlinear detectors. In particular, in channels where the orthogonality of the code sequences is destroyed by multipath, detectors with linear complexity perform much worse than the nonlinear detectors. In this paper, we propose an enhanced multisurface method (EMSM) for multiuser detection in multipath channels. EMSM is an intermediate piecewise linear detection scheme with a run-time complexity linear in the number of users. Its bit error rate performance is compared with existing linear detectors, a nonlinear radial basis function detector trained by the new support vector learning algorithm, and Verdu's optimal detector. Simulations in multipath channels, for both synchronous and asynchronous cases, indicate that it always outperforms all other linear detectors, performing nearly as well as nonlinear detectors. PMID- 17131672 TI - Training recurrent neurocontrollers for robustness with derivative-free Kalman filter. AB - We are interested in training neurocontrollers for robustness on discrete-time models of physical systems. Our neurocontrollers are implemented as recurrent neural networks (RNNs). A model of the system to be controlled is known to the extent of parameters and/or signal uncertainties. Parameter values are drawn from a known distribution. For each instance of the model with specified parameters, a recurrent neurocontroller is trained by evaluating sensitivities of the model outputs to perturbations of the neurocontroller weights and incrementally updating the weights. Our training process strives to minimize a quadratic cost function averaged over many different models. In the end, the process yields a robust recurrent neurocontroller, which is ready for deployment with fixed weights. We employ a derivative-free Kalman filter algorithm proposed by Norgaard et al. and extended by Feldkamp et al. (2001) and Feldkamp et al. (2002) to neural network training. Our training algorithm combines effectiveness of a second-order training method with universal applicability to both differentiable and nondifferentiable systems. Our approach is that of model reference control, and it extends significantly the capabilities proposed by Prokhorov et al. (2001). We illustrate it with two examples. PMID- 17131673 TI - Upport vector machines for nonlinear kernel ARMA system identification. AB - Nonlinear system identification based on support vector machines (SVM) has been usually addressed by means of the standard SVM regression (SVR), which can be seen as an implicit nonlinear autoregressive and moving average (ARMA) model in some reproducing kernel Hilbert space (RKHS). The proposal of this letter is twofold. First, the explicit consideration of an ARMA model in an RKHS (SVM ARMA2K) is proposed. We show that stating the ARMA equations in an RKHS leads to solving the regularized normal equations in that RKHS, in terms of the autocorrelation and cross correlation of the (nonlinearly) transformed input and output discrete time processes. Second, a general class of SVM-based system identification nonlinear models is presented, based on the use of composite Mercer's kernels. This general class can improve model flexibility by emphasizing the input-output cross information (SVM-ARMA4K), which leads to straightforward and natural combinations of implicit and explicit ARMA models (SVR-ARMA2K and SVR ARMA4K). Capabilities of these different SVM-based system identification schemes are illustrated with two benchmark problems. PMID- 17131674 TI - Estimating the number of hidden neurons in a feedforward network using the singular value decomposition. AB - In this letter, we attempt to quantify the significance of increasing the number of neurons in the hidden layer of a feedforward neural network architecture using the singular value decomposition (SVD). Through this, we extend some well-known properties of the SVD in evaluating the generalizability of single hidden layer feedforward networks (SLFNs) with respect to the number of hidden layer neurons. The generalization capability of the SLFN is measured by the degree of linear independency of the patterns in hidden layer space, which can be indirectly quantified from the singular values obtained from the SVD, in a postlearning step. A pruning/growing technique based on these singular values is then used to estimate the necessary number of neurons in the hidden layer. More importantly, we describe in detail properties of the SVD in determining the structure of a neural network particularly with respect to the robustness of the selected model. PMID- 17131675 TI - Solving quadratic programming problems by delayed projection neural network. AB - In this letter, the delayed projection neural network for solving convex quadratic programming problems is proposed. The neural network is proved to be globally exponentially stable and can converge to an optimal solution of the optimization problem. Three examples show the effectiveness of the proposed network. PMID- 17131676 TI - A class of self-stabilizing MCA learning algorithms. AB - In this letter, we propose a class of self-stabilizing learning algorithms for minor component analysis (MCA), which includes a few well-known MCA learning algorithms. Self-stabilizing means that the sign of the weight vector length change is independent of the presented input vector. For these algorithms, rigorous global convergence proof is given and the convergence rate is also discussed. By combining the positive properties of these algorithms, a new learning algorithm is proposed which can improve the performance. Simulations are employed to confirm our theoretical results. PMID- 17131677 TI - Cellular neural field and its convergence analysis. AB - A new continuum model complementary to traditional cellular neural networks is introduced in this note. We consider a cellular neural field formed by infinitely many cellular neurons and modelled by an integrodifferential equation. Then, using LaSalle's invariance principle on Banach space, we show that the field quantity will asymptotically converge to an equilibrium state under the condition that all equilibria of the system are isolated. From the practical sense, the convergence indicates the essential capability of retrieving message from original raw data. PMID- 17131678 TI - An improved neural network model for the two-page crossing number problem. AB - The simplest graph drawing method is that of putting the vertices of a graph on a line and drawing the edges as half-circles either above or below the line. Such drawings are called two-page book drawings. The smallest number of crossings over all two-page drawings of a graph G is called the two-page crossing number of G. Cimikowski and Shope have solved the two-page crossing number problem for an n vertex and m-edge graph by using a Hopfield network with 2 m neurons. We present here an improved Hopfield model with m neurons. The new model achieves much better performance in the quality of solutions and is more efficient than the model of Cimikowski and Shope for all graphs tested. The parallel time complexity of the algorithm, without considering the crossing number calculations, is O(m) for the new Hopfield model with m processors clearly outperforming the previous algorithm. PMID- 17131679 TI - Global asymptotical stability of recurrent neural networks with multiple discrete delays and distributed delays. AB - By employing the Lyapunov-Krasovskii functional and linear matrix inequality (LMI) approach, the problem of global asymptotical stability is studied for recurrent neural networks with both discrete time-varying delays and distributed time-varying delays. Some sufficient conditions are given for checking the global asymptotical stability of recurrent neural networks with mixed time-varying delay. The proposed LMI result is computationally efficient as it can be solved numerically using standard commercial software. Two examples are given to show the usefulness of the results. PMID- 17131680 TI - Kernel classifier construction using orthogonal forward selection and boosting with Fisher ratio class separability measure. AB - A greedy technique is proposed to construct parsimonious kernel classifiers using the orthogonal forward selection method and boosting based on Fisher ratio for class separability measure. Unlike most kernel classification methods, which restrict kernel means to the training input data and use a fixed common variance for all the kernel terms, the proposed technique can tune both the mean vector and diagonal covariance matrix of individual kernel by incrementally maximizing Fisher ratio for class separability measure. An efficient weighted optimization method is developed based on boosting to append kernels one by one in an orthogonal forward selection procedure. Experimental results obtained using this construction technique demonstrate that it offers a viable alternative to the existing state-of-the-art kernel modeling methods for constructing sparse Gaussian radial basis function network classifiers that generalize well. PMID- 17131681 TI - O(log2 M) self-organizing map algorithm without learning of neighborhood vectors. AB - In this letter, a new self-organizing map (SOM) algorithm with computational cost O(log2 M) is proposed where M2 is the size of a feature map. The first SOM algorithm with O(M2) was originally proposed by Kohonen. The proposed algorithm is composed of the subdividing method and the binary search method. The proposed algorithm does not need the neighborhood functions so that it eliminates the computational cost in learning of neighborhood vectors and the labor of adjusting the parameters of neighborhood functions. The effectiveness of the proposed algorithm was examined by an analysis of codon frequencies of Escherichia coli (E. coli) K12 genes. These drastic computational reduction and accessible application that requires no adjusting of the neighborhood function will be able to contribute to many scientific areas. PMID- 17131682 TI - A cross-layer adaptation scheme for improving IEEE 802.11e QoS by learning. AB - In this letter, we propose a cross-layer adaptation scheme which improves IEEE 802.11e quality of service (QoS) by online adapting multidimensional medium access control (MAC)-layer parameters depending on the application-layer QoS requirements and physical layer (PHY) channel conditions. Our solution is based on an optimization approach which utilizes neural networks (NNs) to learn the cross-layer function. Simulations results demonstrate the effectiveness of our adaptation scheme. PMID- 17131683 TI - A low-power current mode fuzzy-ART cell. AB - This paper presents a very large scale integration (VLSI) implementation of a low power current-mode fuzzy-adaptive resonance theory (ART) cell. The cell is based on a compact new current source multibit memory cell with online learning capability. A small prototype of the designed cell and its peripheral block has been fabricated in the AustriaMicroSystems (AMS)-0.35-microm technology. The cell occupies a total area of 44 x 34 microm2 and consumes a maximum current of 22 nA. PMID- 17131684 TI - [Influenza vaccination: NOW!]. PMID- 17131685 TI - [Entecavir. Treatment of chronic hepatitis b virus infections]. PMID- 17131686 TI - [Endocannabinoids as molecular instruments of health promotion]. AB - Endocannabinoids may be a physiological model for our self-healin capacities, since they are part of a complex system of natural auto-regulatory processes. This system has been examined via neurobiology, where the experimental invertebrate model is useful. Endocannabinoids, as well as endogenous morphine, activate constitutive nitric oxide (NO) release, which exerts a variety of positive physiological effects. By doing so, we surmise endogenous stress reduction emerges. Therefore, in the context of endocannabinoid auto-regulation, it seems adequate to speak of "health promotion on a molecular level". The convergence of endogenous auto-regulation on NO pathways critically relies upon common or overlapping neurobiological molecular components, as they are represented by limbic reward and motivation mechanisms. To our knowledge, endogenous auto-regulation--involving deep limbic brain activities--plays a crucial role in successful modern strategies of applied and integrative health promotion. More research, however, is necessary before the different aspects of neurobiological science and clinical medicine in the field of prevention may be integrated extensively and with profound reason. CONCLUSIONS: Successful preventive programs, such as integrative medical stress management, include auto regulative mechanisms on the physiological level. This leads to an interesting research potential, particularly when one considers the long-term effects of applied health promotion and its coupling to motivational neurobiological phenomena. PMID- 17131687 TI - [Sleep disturbances--aspects of obstructive and sleep-related breathing disorders]. AB - Obstructive sleep apnea (OSA) is a wide spread affliction in industrialised countries. OSA has to be distinguished from simple snoring which is socially annoying but physically harmless, since it does not harm the physical health. Patients with OSA complain about disruptive snoring, daytime sleepiness, and loss of intellectual power. OSA is associated with increased mortality. It is possible to distinguish between complete respiratory pauses or apnoea and periods of too shallow breathing or hypopnoea. History taking, clinical examination, recording of daytime sleepiness, and cardio-respiratory sleep studies are necessary to make a proper diagnosis. OSA should primarily be treated, not so the simple snoring. In sleep apneics the treatment focuses on the removal of the sleep-related breathing disturbances and their health-related consequences. Therefore adjuvants or minimally invasive surgical techniques are favoured. Gold standard treatment of OSA is nasally applied continuous positive airway pressure (nCPAP). Alternative treatment modalities are also used. As conservative approaches oral appliances and conditioning (to avoid supine sleeping position) show promising results. Surgical treatment complies with the severity of the disease and with the anatomical findings. A variety of surgical approaches to the soft palate, the base of tongue, the tonsils, and the upper and lower jaws have been described. PMID- 17131688 TI - [Lutein as nutritional supplement]. PMID- 17131689 TI - [Attention deficit disorder with hyperactivity (ADHD). Continuing symptom control with medium term methylphenidate treatment]. PMID- 17131690 TI - ["New" coronaviruses are the cause of respiratory diseases]. PMID- 17131691 TI - [Prevention of atherothrombosis with low doses of acetylsalicylic acid]. PMID- 17131692 TI - [Combination of acetylsalicylic acid with dipyridamole is better than acetylsalicylic acid monotherapy]. PMID- 17131693 TI - [Cardiovascular and respiratory complaints from fine particles]. PMID- 17131694 TI - [Treatment of phantom pains]. PMID- 17131695 TI - [New mechanism of action RANKL-antibodies reduce bone loss]. PMID- 17131696 TI - [Symptomatic gallstone disease: an indication for surgery]. AB - Three patients, men in the ages of 58, 66 and 56 years, respectively, had experienced 'warning colics' a considerable time before gallstone complications or severe recurrent colic. Ultrasonographically proven gallstones had not led to cholecystectomy. The 58-year-old man died of sepsis due to infected pancreatic necrosis; the other men underwent laparoscopic cholecystectomy, after which they recovered fully. Approximately 10-5% of the adult Dutch population have gallstones, but only 10% will develop symptoms. The annual risk for developing complicated gallstone disease is 1-2% in asymptomatic gallstone carriers. Of patients admitted with complicated gallstone disease, 58% have had prior 'warning colics'. Complicated gallstone disease can be prevented by timely treatment after recognition of warning colics. Cholecystectomy is indicated in patients with intermittent upper-abdominal pain and proven gallstones or sludge. PMID- 17131697 TI - [Freezing umbilical-cord blood and bone marrow for one's own use: present-day quackery?]. AB - In the Netherlands, the practice of private freezing and banking of umbilical cord blood is increasing. In a questionnaire, Dutch midwives and gynaecologists were asked about their attitude towards cord-blood collection if asked to perform this after delivery. The response rate was 35% (125/356) and 71% (71/100), respectively. Two-thirds of those asked responded that they would comply. The most common application of cord blood is in the treatment of (malignant) blood disorders. The use of autologous cord blood is, however, often not the best choice for treating leukaemia in young children and the number of stem cells is often too low in a single-cord blood sample to treat older children and adults. Although frequently suggested in the lay press, there is no proven effect in other indications, such as amyotrophic lateral sclerosis, multiple sclerosis and myocardial infarction. Information on therapeutic applications of cord blood from companies with commercial interests is leading to the exploitation ofpregnantwomen. The government should consider limiting this practice and prohibiting the activities of these companies in the Netherlands pending scientific evidence for their claims. PMID- 17131698 TI - [Urinary and faecal incontinence in the elderly at home]. AB - Urinary and faecal incontinence are a growing problem in the Dutch elderly population. Recent figures from general practice suggest an average prevalence in the over-60-year-olds of 19 and 6%, respectively. Due to an accurate system of continuous monitoring and a very high response on a non-validated but clearly worded and short questionnaire, the reliability of these figures is most probably high. The research article is one in a series on incontinence from the same research group that studied the practical implications of urinary incontinence in the elderly. Topics published so far concerned - among others - a systematic review of the efficacy of available interventions for urinary incontinence and an in-depth study of the barriers felt by GPs regarding their familiarity with and frequency of use of these interventions. The results of these studies can easily be translated for postgraduate education. Sadly, the magnitude and quality of the knowledge base for the treatment of faecal incontinence still lags behind. PMID- 17131699 TI - [Preventive resection of hereditary diffuse gastric cancer]. AB - Hereditary diffuse gastric cancers are rare, accounting for at most 1-3% of gastric cancers. It can be caused by a mutation in the tumour-suppressor gene CDH1. A healthy person carrying a CDH1 mutation has a cumulative risk of developing gastric cancer of 70-80%. In most cases, gastric cancer is detected before the age of 40 years. The effectiveness of screening for hereditary diffuse gastric cancer or early detection with twice-yearly upper GI endoscopy with blind biopsies is highly questionable. Given the poor prognosis of patients with hereditary diffuse gastric cancer, prophylactic gastrectomy can be considered an option for patients with a CDH1 mutation. It is recommended that the supervision, screening and possible preventative gastrectomy for hereditary diffuse gastric cancers are handled by a multidisciplinary team in a specialised centre. PMID- 17131700 TI - [The practice guideline 'Volume suppletion in critically-ill neonates and children up to the age of 18 years' of the Dutch Paediatric Association]. AB - Hypovolaemia is the most common cause of circulatory failure in children. Treatment consists of volume suppletion with a crystalloid or colloid solution; which agent is the best in children is not clear. This evidence-based practice guideline formulates recommendations as to which fluid should be used for volume suppletion in critically-ill neonates and children up to the age of 18 years with hypovolaemia. Before the guideline development first-choice fluid for volume resuscitation was in 50% a colloid and in 50% a crystalloid solution for both neonatologists and paediatric intensivists. The neonatologists used human albumin as a priority, and the paeditric intensivists predominantly used a synthetic colloid. The guideline was developed on the basis of a comprehensive search and analysis of the literature according to the principles of evidence-based guideline development. The recommendations were formulated by a committee based on evidence from the literature and, when evidence from the literature was insufficient, on consensus after discussion in the committee. Since colloids are much more expensive than crystalloids and can give an anaphylactic reaction, their added value over crystalloids must be proven. In sick neonates and children, insufficient clinical trials have been done to reach the conclusion that colloids are more effective than crystalloids in hypovolaemia. A number of meta-analyses in adults revealed excess mortality in the group treated with albumin, but one recent, large, randomised study showed no difference in mortality. No added value could be demonstrated for the administration of synthetic colloids. On the basis of data from the literature and considerations regarding the applicability of evidence in adults to children and neonates, the side effects of resuscitation fluids, pathophysiology and costs, the first-choice fluid for neonates and children with hypovolaemia is isotonic saline. Albumin should not be used for the treatment of hypovolaemia. The volume to be administered and the infusion rate depend on the severity of the hypovolaemia and should be determined on an individual basis. PMID- 17131701 TI - [New medications; ziconotide]. AB - Ziconotide is a synthetic analogue of a peptide found in the poison of the marine snail Conus magus. Ziconotide blocks N-type calcium channels, which play an important role in the transmission of pain signals in the dorsal ganglia of the spinal cord. The drug is indicated for 'severe chronic pain' and is administered intrathecally. PMID- 17131702 TI - [Diagnostic image (298). A woman with a gingival swelling]. AB - A woman aged about 25 years had a local swelling of the gums of her teeth during pregnancy: epulis gravidarum. PMID- 17131703 TI - [Prevalence of urinary and faecal incontinence among community-dwelling elderly patients in Nijmegen, The Netherlands, January 1999-July 2001]. AB - OBJECTIVE: To evaluate the prevalence of urinary, faecal and double incontinence in community-dwelling elderly patients. DESIGN: Cross-sectional population-based survey. METHOD: By means of a postal questionnaire, data were collected in the period January 1999-July 2001 from patients aged 60 and over from 9 general practices associated with the academic general practitioner registration network of the St Radboud University Medical Centre (the Nijmegen Monitoring Project). Excluded were patients living in a home for the elderly, as well as patients with dementia, patients who were too ill to participate and patients with a catheter. RESULTS: Of the 5278 patients who received a questionnaire, 4650 (88%) returned it. 885 (19%) respondents had involuntary loss of urine twice a month or more, 299 (6%) had involuntary loss of faeces and 153 (3%) had both. The prevalence of urinary, faecal and double incontinence increased with age in both men and women, in men especially in the age group > 80 years. Urinary incontinence was more prevalent in women (29%) than in men (9%). The prevalence of faecal incontinence showed no sex differences (women: 6%; men: 7%), but the loss of slimy faeces occurred twice as often in men as in women (60% versus 29%). Double incontinence was also more or less equally prevalent in men and women (men: 2%; women: 4%), except in the age group 65-74 years (men: 1%; women: 4%). CONCLUSION: Especially urinary, but also faecal incontinence was common in the community-dwelling elderly. The prevalence increased with age. Because of ageing of the population and the increasing life expectancy in the next decennia, the prevalence of incontinence can be expected to increase considerably. PMID- 17131704 TI - [Protons and ions in the treatment of cancer; a systematic review of the literature]. AB - OBJECTIVE: To provide an overview of the present role of proton and ion therapy, also referred to as 'charged particle therapy', in the treatment of cancer. DESIGN: Systematic literature study. METHOD: Systematic electronic searches were carried out in 12 databases according to the Cochrane Collaboration criteria, without restriction as to year of publication or study design. Manual searches of bibliographies and journals were also performed. The inclusion criteria were: at least 20 patients and a follow-up of at least 2 years. In addition, experts on the subject were consulted by correspondence for their opinion. RESULTS: The search identified 36 relevant articles on proton therapy and 15 on ion therapy. Based on prospective and retrospective studies, proton irradiation emerged as the treatment of choice for ocular tumours, chordomas and skull-base tumours. For prostate cancer, the results were comparable with the best results of photon therapy. Ion therapy was still in an experimental phase. CONCLUSION: According to the current literature, proton therapy is looked upon as the preferred treatment modality for certain rare tumours, such as ocular tumours, chordoma, and skull base tumours. However, charged particle therapy as a whole, and especially ion therapy, is not supported as the treatment of choice for cancer by published evidence. Nevertheless, the potential theoretical benefit of this treatment is great. PMID- 17131705 TI - [Endocarditis due to meticillin-resistant Staphylococcus aureus originating from pigs]. AB - A 63-year-old woman with a kidney transplant was admitted with endocarditis caused by meticillin-resistant Staphylococcus aureus (MRSA). Once her antibiotic therapy had been adjusted to the sensitivity-pattern of the bacterial strain she recovered, without the need for surgical intervention. The isolated S. aureus was typed by multi-locus sequence typing as sequence type 398, a MRSA-strain that has recently been isolated from a high percentage of Dutch pigs. This is the first report of a life-threatening infection with this pig MRSA. This strain is genetically different from the globally dispersed nosocomial MRSA-strains, and also from the strains that have been epidemic for several years in the USA as the causative agent ofcommunity-acquired skin infections. The Dutch Working Group on Infection Prevention (WIP) has recently adjusted its guidelines to halt further spread of this strain, and advises that the population at risk (pig breeders, slaughterhouse personnel and veterinarians) be held in isolation when hospitalised until MRSA colonisation has been excluded. The patient described here, however, did not belong to this population at risk. PMID- 17131706 TI - [Rapid prenatal diagnosis of chromosomal abnormalities; limitations and possibilities]. PMID- 17131707 TI - [Rapid prenatal diagnosis of chromosomal abnormalities; limitations and possibilities]. PMID- 17131708 TI - [Legionella outbreak in Amsterdam: a cooling tower as the source]. PMID- 17131709 TI - Tooth surface loss--a challenge to oral health. PMID- 17131710 TI - The role of erosion, abrasion and attrition in tooth wear. AB - There is increasing clinical awareness of erosion of enamel and dentine by dietary acids and the consequent increased susceptibility to physical wear. Enamel erosion is characterized by acid-mediated surface softening that, if unchecked, will progress to irreversible loss of surface tissue, potentially exposing the underlying dentine. In comparison, dentine erosion is less well understood as the composition and microstructure are more heterogeneous. Factors which affect the erosive potential of a solution include pH, titratable acidity, common ion concentrations, and frequency and method of exposure. Abrasion and attrition are sources of physical wear and are commonly associated with tooth brushing and tooth-to-tooth contact, respectively. A combination of erosion and abrasion or attrition exacerbates wear; however, further research is required to understand the role of fluoride in protecting mineralized tissues from such processes. Abrasive wear may be seen in a wide range of patients, whereas attritive loss is usually seen in individuals with bruxism. Wear processes are implicated in the development of dentine hypersensitivity. Saliva confers the major protective function against wear due to its role in pellicle formation, buffering, acid clearance, and hard tissue remineralization. This review focuses on the physiochemical factors impacting tooth wear. PMID- 17131711 TI - Surface microhardness changes, enamel fluoride uptake, and fluoride availability from commercial toothpastes. AB - OBJECTIVE: The aim of the present study was to evaluate the ability of a new fluoride-containing dentifrice to protect surface-softened enamel against further erosive challenges in an in vitro cycling model, and to relate any effects to enamel fluoride uptake (EFU) and free fluoride. METHODOLOGY: Human enamel specimens were subjected to a daily cycling regimen comprising: three two-minute treatments; five two-minute challenges using 1% citric acid pH 3.8; and remineralization in a mixture of human saliva and mucin-containing artificial saliva. Surface microhardness (SMH) was measured at baseline, 10, and 20 days, and the fluoride content of biopsied specimens determined at 20 days. EFU studies were based on method #40 described in the United States Food and Drug Administration (FDA) testing procedures. Free-fluoride availability was determined from slurries of one part toothpaste plus three parts deionized water. RESULTS: SMH showed that a 1150 ppm NaF test dentifrice protected enamel specimens greater than Crest Cavity Protection (1100 ppm NaF) and a fluoride-free placebo at both 10 days and 20 days (p < 0.05). The fluoride content of specimens treated with this prototype was higher than either Crest or the placebo. SMH for a 1450 ppm NaF test dentifrice was greater than for Elmex Sensitive (1450 ppm amine F) and placebo at 10 days, while both products were greater than the placebo at 20 days. The fluoride content of specimens treated with this test dentifrice was higher than Elmex Sensitive, which was higher than placebo. The fluoride uptake seen in the cycling model correlated for the NaF dentifrices with a standard EFU procedure. Different EFU results for a series of commercial dentifrices demonstrated that EFU is not necessarily a function of free-fluoride availability. CONCLUSION: This study demonstrated that fluoride dentifrices can increase the protection of enamel against an erosive challenge in vitro, and that the increased protection correlated with fluoride uptake. The fluoride uptake seen in the cycling model correlated with a standard FDA EFU procedure for the NaF dentifrices. The present studies demonstrate the importance of formulation effects on driving performance in in vitro models. PMID- 17131712 TI - In vitro microhardness studies on a new anti-erosion desensitizing toothpaste. AB - OBJECTIVE: To evaluate in vitro the efficacy of a new anti-erosion desensitizing toothpaste to inhibit enamel surface softening by a dietary acid, and promote re hardening of artificial erosive lesions. METHODOLOGY: The ability of the toothpaste to inhibit formation and promote repair of erosive lesions in human enamel has been investigated. In an enamel surface softening study, sound human enamel was pre-treated with one of four toothpaste slurries for two minutes, before exposure to 1.0% citric acid, pH 3.8, for a total of 30 minutes. The surface microhardness (SMH) of the specimens was determined at baseline and at 10 minute intervals using a Struers Duramin-1 microindentor. In an enamel re hardening study, the erosive lesions were prepared by exposure of the specimens to 1.0% citric acid, pH 3.8, for 30 minutes. After two minutes treatment with a toothpaste slurry, lesion repair was monitored by SMH after 4, 24, and 48 hours incubation in artificial saliva. This remineralizing phase was modified by the addition of an aliquot of the relevant toothpaste slurry, to mimic in vivo carryover of the formulation. RESULTS: The new test formulation, Elmex Sensitive, and Colgate Sensitive exhibited statistically significant inhibition of citric acid-mediated enamel surface softening versus a fluoride-free placebo at all time points. The test toothpaste gave statistically superior protection against the erosive challenge compared to Elmex Sensitive and Colgate Sensitive after 20- and 30-minute exposures. In the remineralization studies, erosive lesions treated with the test toothpaste exhibited statistically superior re-hardening versus lesions treated with Elmex Sensitive and Colgate Sensitive after 24- and 48-hour incubation in the artificial saliva. Lesions treated with Elmex Sensitive re hardened to a statistically significant extent versus the fluoride-free placebo toothpaste. A re-hardening study, in which a series of the new toothpaste-base formulations containing increasing concentrations of NaF were evaluated, showed a clear fluoride dose response. CONCLUSION: The present microhardness studies show that treatment with fluoride-containing toothpastes helps protect sound enamel from acid-mediated surface softening, and promotes re-hardening of erosive lesions. The new test toothpaste exhibited statistically superior efficacy to Elmex Sensitive and Colgate Sensitive in both in vitro models. PMID- 17131713 TI - The stain removal performance of a new anti-hypersensitivity dentifrice. AB - OBJECTIVE: To investigate the stain removal potential in vitro of a new anti hypersensitivity dentifrice. The dentifrice contains a low level of abrasive, the zwitterionic surfactant cocamidopropyl betaine, and potassium nitrate. It has been developed to be as gentle as possible to tooth surfaces and oral soft tissues, while effectively treating dentinal hypersensitivity. METHODOLOGY: The Relative Dentine Abrasivity (RDA) method was used to measure abrasivity. The Pellicle Cleaning Ratio (PCR) and the Natural Extrinsic Stain Removal (NESR) methods were used to test stain removal performance against suitable controls. RESULTS: The RDA value for the formulation was 34 +/- 2 (Mean +/- S.E.). The PCR value was 46 +/- 4, comparable with Elmex Sensitive. The NESR test, which has previously been shown to give better clinical correlation than the PCR, demonstrated that the new formulation gave superior stain removal performance compared with both Sensodyne MultiCare and a conventional nonsensitivity formulation, and similar stain removal performance to Elmex Sensitive. CONCLUSION: The new formulation has been shown, in these studies, to combine low abrasivity with an in vitro stain removal performance comparable with that of benchmark marketed pastes. PMID- 17131714 TI - Evaluation of a desensitizing test dentifrice using an in situ erosion remineralization model. AB - OBJECTIVE: To evaluate, in an in situ remineralization model, the ability of a low abrasion fluoride dentifrice containing potassium nitrate to enhance the remineralization of enamel that was previously subjected to an in vitro dietary erosion challenge. METHODOLOGY: Thirteen subjects completed a single-blind (to specimen analyst) crossover design study with four randomly assigned dentifrice treatments: placebo dentifrice (0 ppm F; PD); dose response control dentifrice (250 ppm F; DD); clinically tested fluoride dentifrice (1100 ppm F; FD); and test dentifrice (1150 ppm F + 5% KNO3; TD). Each subject wore a palatal appliance holding eight bovine enamel blocks that were previously exposed for 25 minutes to an in vitro erosive challenge with grapefruit juice. Surface microhardness (SMH) was determined prior to the erosive challenge (baseline), after the in vitro erosive challenge, after in situ remineralization, and after a second in vitro erosive challenge. Statistical analyses included ANOVA and pair-wise comparisons between treatments, testing at a 5% significance level. RESULTS: The mean percent SMH recovery (SD) was 20.4 (7.7)a for PD; 27.4 (5.9)b for DD; 29.5 (8.9)bc for FD; 33.4 (6.7)c for TD. The mean percent net erosion resistance (SD) was -57.7 (10.8)a for PD; -41.3 (11.6)b for DD; -28.7 (9.7)c for FD; -23.6 (7.3)c for TD. Different superscript letters following the means imply statistically significant differences between groups for each response. CONCLUSION: The test dentifrice was shown to significantly enhance the remineralization of enamel previously subjected to an erosion challenge. PMID- 17131715 TI - Preventing medication errors in hospitals through a systems approach and technological innovation: a prescription for 2010. AB - Medication errors in hospital settings are considered both widespread and costly to the American healthcare system; yet, it is tractable to available solutions. This article offers a novel prescription for the problem that could be implemented by 2010. It consists of a systems approach--failure mode effects analysis (FMEA)--in combination with emerging technologies, such as a decision support system (DDS) with integrated real-time medical informatics, electronic medical records (EMR), computer physician order entry (CPOE), bar coding, automated dispensing machines (ADM), and robotics. Cost and benefit analysis reveals that this proposed integrated solution will radically reduce medication errors in hospitals and save the lives of thousands of Americans who frequent such facilities on an annual basis, as well as reduce healthcare costs. PMID- 17131716 TI - Strategic human resource management issues in hospitals: a study of a university and a community hospital. AB - The human factor is central to healthcare, yet its proper management has remained beyond the reach of healthcare organizations. This qualitative study examines strategic human resource management (HRM) issues in a university and a community hospital. The findings indicate that the two hospitals lacked a clear understanding of their strategic intent and objectives; as a result, their human resource (HR) practices lacked coherence and direction. Whereas the community hospital understood the interrelationship between culture and HRM, the university hospital did not. Moreover, the university hospital showed only a modest understanding of competencies needed in managing HR function, which hampered its ability to identify competent HR managers and employees. The community hospital made significant gains in the past few years in managing its culture and people by recruiting a competent HR manager. The relationship between HR practices and clinical outcomes was much less clear in the university hospital than it was in the community hospital. PMID- 17131718 TI - Virtue ethics: worth another look. PMID- 17131717 TI - Perceptions of hospital CEOs about the effects of CEO turnover. AB - Empirical evidence is scarce on chief executive officer (CEO) turnover in U.S. hospitals, with potentially serious implications for many of these organizations. This study, based on a nationwide survey of CEOs at non-federal general surgical and medical community hospitals conducted in the spring of 2004, reports the perceptions of hospital CEOs regarding the circumstances and impact of CEO turnover on U.S. hospitals. In the opinion of the respondents, the impact includes competitors taking advantage of turnover by luring employees and physicians away from the target hospital, significantly increasing the likelihood of other senior executives leaving the hospital, and many of the important strategic activities being delayed or cancelled altogether. Interestingly, the perceptions of CEOs regarding the effects of turnover do not seem to differ regardless of voluntary or involuntary circumstances of turnover. However, there is a notable bias in emphasizing the perceived negative implications of respondents' own departures and allegedly positive effects of their predecessors' departures. PMID- 17131719 TI - Overall outpatient satisfaction and its components: perceived changes at the Huntington VA Medical Center over five years. AB - The number of veterans obtaining healthcare services at Huntington Veteran's Administration Medical Center (HVAMC) had significantly increased over the five year span from fiscal year 2000 through fiscal year 2004. The authors' purpose in this study was to determine changes in outpatient satisfaction levels at HVAMC over that five-year period. The authors assessed the following specific measures of patient satisfaction: courtesy, access, patient preferences, coordination of care, education and information, emotional support, overall quality, pharmacy services, and overall satisfaction. Of the 17,000 patients meeting the criteria, 630 were randomly selected; 195 patients completed surveys, resulting in a 31% response rate. Results demonstrated an overall significantly higher level of patient satisfaction than five years previously. On the basis of these results, the medical center administration can probably conclude that any process he improvements made over the five-year period have been successful. PMID- 17131720 TI - [Epidemiological analysis of posttraumatic cervical spine injury]. AB - The purpose of this study was epidemiological analysis of trauma causes and trauma effects including age and sex. 112 patients after cervical spine trauma, admitted in Emergency Room of Self-Financing Public District Hospital of Trauma Surgery at Piekary Slaskie were studied retrospectively. The following parameters were estimated: age, sex, cause of injury, type and localization of pathology. The mean age was 35.4 years. The largest age group consisted of patients 25-45 years old. Males predominated over females in all age groups. The most common cause of cervical spine injury was traffic accident with a patient as a car driver or as a car passenger (45.5%) The most common pathology was luxation and subluxation (36.6%). Luxations and subluxations were most often localized on C1 C2 level (50.6% of all luxations), and on C5-C6 level (19.8% of all luxations). Fractures were most often localized in C5 vertebra (24.8% of all fractures). These results suggest that causes and results of trauma and participation of both sex have changed. PMID- 17131721 TI - [The results of decompression and anterior lumbar interbody fusion with the use of interbody cages for the treatment of degenerative lumbar spondylolisthesis]. AB - In this paper we reviewed 28 patients who had been treated surgically for lumbar degenerative spondylolisthesis. They were operated between 1998-2003. The decompression and anterior lumbar interbody fusion with the use of interbody cages, was performed in all of them. The outcome was assessed using rating system of Prolo and VAPS. The disc height, degree of slippage and segmental lordosis were measured, on the radiographs, before surgery, after 6 weeks and at the time of final follow-up. In all cases spinal fusion was achieved. The disc height, degree of slipage and segmental lordosis were improved and these results were stable in time. A significant decrease in radicular pain and low back pain were seen but the relation between clinical and radiological autcomes was not observed. PMID- 17131722 TI - [The prognostic factors in perthes disease]. AB - The purpose of this study was to evaluate the role of selected factors which may play role in the estimation of late result during Perthes disease and which one can be eliminated during treatment period. The study population consisted of 311 patients (50 patients with bilateral disease, 361 hips joint) who had reached skeletal maturity at last follow up. All hips were treated by containment methods (bed rest and traction in abduction, Petri cast, brace, varus osteotomy, Salter osteotomy and shelf acetabuloplasty). Both clinical and radiological parameters (taken during fragmentation stage) were included in our study: gender, age at the onset, hip joint abduction, type of treatment, extend of the femoral head necrosis according to the Herring and Catterall classification, LLD, premature growth plate arrest, ATD and ATD index, lateral acetabulum shape - type I - normal, concave lateral acetabulum margin, type II--flat, horizontal and type III -convex, sloping, femoral head subluxation, femoral head sphericity disturbance according to the Mose, risk factors according to the Catterall, Wiberg angle, Eyre-Brook index. Long-term results were evaluated according to the Stulberg classification and were divided into two groups: good result--Stulberg group 1 or 2 and poor and bad result--Stulberg group 3, 4 or 5. The unilateral and bilateral Perthes disease was estimated separately. The statistical analysis revealed in unilateral Perthes disease that next risk factors may lead to poor or bad late result: age at the onset 9 and more years, group 3 or 4 according to Catterall and group C according to Herring classification, type III lateral acetabulum shape, hip joint abduction less than 10 degrees, premature growth plate arrest, ATD index less than 0.8, femoral head subluxation more than 20%, femoral head sphericity disturbance more than 5mm, Gage sign, calcification laterally to the femoral head (sensitivity 85.0%, specificity 98.7%). Bilateral Perthes disease: age at the onset over 6 years, group 3 or 4 according to Catterall and group C according to Herring classification, hip joint abduction less than 10 degrees and calcification laterally to the femoral head (sensitivity 81.8%, specificity 92.3%). The statistical analysis showed which clinical and radiological factors may play an important role in estimation of late results during Perthes disease. Additionally during fragmentation stage of Perthes disease we have possibility to correct or eliminate some of the risk factors - femoral head subluxation, lateral acetabulum shape and hip joint abduction and improve the late result. PMID- 17131723 TI - [Chiari pelvic osteotomy in the treatment of hip dysplasia in adults]. AB - INTRODUCTION: [corrected] The aim of the study is early results evaluation of Chiari pelvic osteotomy. MATERIALS AND METHODS: In the years 1997-2003 36 patients (26 women and 10 men) in the age from 16 to 50 years old were operated on because of hip joint dysplasia. The indications for operative treatment were: pain, age less than 50 years, insufficient femoral head coverage without evidence of arthritic changes on X-ray. RESULTS: There were no intraoperative and postoperative complications. Harris Hip Score improved from 85 points before operation (from 82 to 90 pts.) to 95 points (from 92 to 100 pts.) after operative treatment. There was pain relief in most of the patients after osteotomy. Osteotomy healing was seen on X-ray examination usually after 6-12 weeks in 33 patients. Delayed osteotomy union till 6th postoperative month without influence on clinical hip improvement was seen in 3 patients. The increase in Wiberg CE angle from average 17.2 degrees (from 3 to 33 degrees) before operation to 44.9 degrees (from 19 to 78 degrees) after operation was statistically significant p< 0.00045. There was no loss in osteotomy correction. CONCLUSION: Supraacetabular osteotomy decreases pain, and increases hip function. Improvement in hip biomechanics and increase in femoral head coverage may lead to decrease in development of hip joint arthritic changes and time preserve before total hip arthroplasty. PMID- 17131724 TI - [Dislocations hip arthroplasty]. AB - The authors present analysis of dislocation following hip arthroplasty based on their own clinical material of 1985-2005 year. It contain 2712 cases of total hip arthroplasty, 2171 (80.1%) cases were cemented including 64 (2.4%) cases of revised arthroplasty, 541 (19.9%) cases were cement-free stabilized including 7 (0.2%) cases of revised arthroplasty. Obtained outcomes of analysis permit to make assertion that dislocation following hip arthroplasty occurred during first three month after surgery, more frequently in case of revised arthroplasty and following fractures of femoral neck. Requirement of success is appropriate steady of implant and right done supervision of rehabilitation with learned necessary motoric behaviors. PMID- 17131725 TI - [Wear and periprosthetic osteolysis of modular, hemispheric acetabular cups]. AB - Osteolysis associated with polyethylene wear has become one of the most prevalent complications associated with uncemented modular, hemispherical cups. Sixty-five consecutive total hip arthroplasties (ABG i, Howmedica, Stryker) were followed 6 12 years. Cumulative survivorship for the cup was 55.7% after 10 years. There were 20 cups revisions because of polyethylene wear and periprosthetic osteolysis (14 cups) or cup loosening (6 cups). Stability was assessed intraoperatively, 14 cups were revised, whereas 6 new polyethylene cups were cemented into stable metal-back of acetabular component. The average annual wear of revised cups was 0.32 mm. The annual wear of not revised cups was 0.12 mm. The differences were statistically significant (p = 0.002). The mean area of osteolysis was 472 mm2 (SD 257 mm2). There was no significant correlation between wear and area of osteolysis. There was significant correlation R = 0.54, p = 0.014) between time to revision and area of osteolysis. There were no significant differences of wear of polyethylene inlay or area of osteolysis between stable and unstable acetabular cups. PMID- 17131726 TI - [Transdifferentiation of chondrocytes into osteogenic cells]. AB - Hypertrophic chondrocytes, commonly considered as terminal cells designated to apoptotic elimination in the model of endochondral osteogenesis, are accordingly to the new concept based on histochemical, immunohistochemical, biochemical and cytological analysis, able to switch their metabolism and enter the osteoblastic differentiation path. According to this concept, some osteocytes in model of endochondral osteogenesis are derivative of hypertrophic chondrocytes. Also non hypertrophic chondrocytes are able to transdifferentiate toward osteogenic cells, and the bone formed by such mechanism is termed "transchondroid bone". PMID- 17131727 TI - [Management of soft tissue defects in the region of finger PIP joints by means of pedicular skin flaps taken from abdomen]. AB - Management of soft tissue defects in the region of finger PIP joints makes a serious operative problem. Commonly recommended skin plastics require a wide experience in hand surgery. There is also lack of a comprehensive doctrine of surgical therapy depending on the extent of the mutilations. The paper presents the results of treatment of 17 mutilated fingers of 11 patients (8 men, 3 females) treated over the period of 2 years (2002-2004). The method applied was pediculated skin flaps taken from an abdomen. According to the extent of lesions three groups of patients were created. Eight patients were treated as emergencies, three appeared from 4 to 11 weeks after the accident. Flaps were cut off after 20.4 days on average. The secondary plastic was performed after the following 9.5 weeks. In 2 patients tendon plastics were performed. Fingers were mobilized during the whole period of the therapy. No complications in healing of the flaps were observed. The functional results were assessed 6 months on average after the primary operation. The average TAM and TPM were evaluated as follows: group I (no, or minor tendon lesions) TAM--186 degrees, group II. PMID- 17131728 TI - [Intramedullary locked nailing in the treatment of proximal femoral fractures]. AB - INTRODUCTION: The aim of the study is evaluation of results of operative treatment the proximal femoral fractures with intramedullary locked nailing. MATERIAL AND METHOD: In years 1996-2004 44 patients were treated because of proximal femoral fractures with closed reduction and stabilization with intramedullary locked nailing. There were 15 women and 29 men in average age 60 years (18-95 years). There were 2 femoral neck fractures and 42 peritrochanteric fractures. The fracture of femoral shaft in 2 patients accompanied the fracture of proximal part of femur. The morphology of fractures was estimated according to AO classification. Clinical results were evaluated with Harris hip score (HHS). RESULTS: The average follow up is 8 months (from 6 to 24 months). There was good reduction of 27 fractures on postoperative radiograms. The average 15 degrees of varus lack of reduction was noted in 17 peritrochanteric fractures (from l0 to 35 degrees). There were mainly 31.A.3 unstable fractures according to AO classification. 3 patients died during three postoperative months from causes not connected with operative treatment. 39 fractures united in the period from 10 to 16 weeks. Delayed union--after 6 months occurred in 2 persons. Fatigue fracture of intramedullary rods occurred to both of this patients. The average Harris Hip Score was 86 points (from 70 to 100 points) after union of fractures. CONCLUSIONS: (1) The use of intramedullary locked nailing in the treatment of fractures of proximal part of femur leads to union without additional immobilization. (2) Closed intramedullary locked nailing in the treatment of unstable intertrochanteric fractures is connected with the risk of varus malalignment of fragments, which does not delay union, but decreases functional outcome. (3) Proper operative technique decreases the number of intra- and postoperative complications. PMID- 17131729 TI - [Radiometric assessment of wrist angle values, linear parameters of the forearm and wrist ratios]. AB - With the device of our own invention suitable for static X-ray examination of the wrist 12 radiographic parameters were evaluated. 100 radiograms regarded normal were analyzed, obtained as a comparative in unilateral wrist trauma patients group. Age of the patients ranged from 18 to 60 years. No comparative studies in regard to sex were performed. Obtained data were statistically analyzed. Subsequent values of the radiometric parameters were obtained: scaphoid-lunate angle (SL) 48.61 degrees, radio-lunate angle (RL): -0.83 degrees, palmar inclination of the distal radius metaphysis (RI): 25.96. The most significant linear parameters measured: ulnar length 0.18 mm, ulnar transposition (UT): 0.339 mm. PMID- 17131730 TI - [Proximal humerus fractures analysis of treatment and rehabilitation outcomes]. AB - The authors make evaluation of functional outcomes proximal humerus fractures depending on applied method using Constant-Murley score in own modification. The clinical material of 1980-2004 years were 93 cases, 41 male (44.1%) and 52 female (55.9%) aged between 28-81 years. The operative treatment was employed in 52 cases (45.1%), nonoperativ in 18 cases (19.4%) and functional treatment using direct traction by olecranion in 33 cases (35.5%). Obtained outcomes based on Constant-Murley score in own modification notice limitation range of motion injured joint, reduction of muscle streught in shoulder rim and decrease of physical activity independently used method of treatment. PMID- 17131732 TI - Business challenges to our professionalism. PMID- 17131731 TI - [Nonunion of the lateral humeral condyle--operative treatment, case report]. AB - The nonunion of the lateral humeral condyle is the most important complication in course of the treatment of the fractures lateral humeral condyle. The cause of the occurrence is the articular liquid penetration to the fracture site, minor blood supply of the fractured fragment based only on the vessels penetrating from the metaphysis, insufficient immobilization period and fragment relocation. The operative treatment of a condyle nonunion previously was performed cautiously because of the high risk of the condyle necrosis and the mild nature of the deformity. The elbow with nonunion condyle is usefull and satisfactory even after bringing increasing valgus deformity and a high risk of the ulnar nerve neuropathy into consideration. A 7-YEAR-old boy with a condyle nonunion was treated operationaly. Operative procedure shouldn,t be prolonged for more than a year after the trauma because of the increased deformity and condyle remodelling. A Tahdjian technique was used. The operation focused on an intraarticular approach with an olecranon ostotomy for the avoidance of the vessels penetrating from the condyle metaphysis. After debridgement and cortical grafts filling of the nonunion site a Kirschner fixation combined with olecranon wire loop fixation were performed. Satisfactory results were achieved in a form of fragments union within six weeks of the surgery and the total range of motion of the elbow joint within 6 months of the operation. During the treatment no signs of the neurovascular complications were observed. PMID- 17131733 TI - Coaptation splinting for humeral shaft fractures in adults and children: a modified method. AB - Coaptation splinting is an accepted form of treatment for humeral shaft fractures in both children and adults. However, application may be difficult when working alone or with an uncooperative patient. We describe a modified method, involving supplies easily found in most emergency departments and cast rooms, that mitigates these problems. PMID- 17131734 TI - A simple and accurate method for determining leg length in primary total hip arthroplasty. AB - Reconstruction of appropriate leg length is an important part of soft-tissue balance in total hip arthroplasty (THA). Leg length discrepancy (LLD) is one of the more common reasons for litigation after otherwise successful THA. The purpose of the study reported here was to analyze the accuracy of using preoperative templating and intraoperative referencing of the well leg to determine postoperative leg length in unilateral primary THA. Seven-hundred primary THAs performed at an institution by 3 surgeons were randomly selected from a computerized database. Cases with significant bilateral disease, congenital dysplasia, acute fracture, or previous surgery or without complete preoperative and postoperative radiographs were excluded. Three reviewers used a standardized method to measure preoperative and postoperative LLD. Included in the review were 410 THAs. Mean postoperative LLD was 3.9 mm lengthening (SD, 7.5 mm). In 20 THAs (4.9%), lengthening was more than 15 mm. Lengthening was more than 20 mm (maximum, 22 mm) in 4 THAs (1%). Of the 20 THAs with LLD of more than 15 mm, 14 involved hips that were longer preoperatively. Thirteen of these hips were reconstructed to within 10 mm of preoperative LLD. Only 2 patients with radiographic LLD of more than 15 mm perceived LLD. There were no differences in gender, height, weight, or body mass index. This method of preoperative templating and referencing the well leg intraoperatively is an inexpensive, reliable, and accurate method for determining leg length in primary THA and has few significant radiographic or clinical outliers. PMID- 17131735 TI - Blood management in total joint arthroplasty. AB - Blood loss associated with total joint arthroplasty can be substantial. Various techniques for dealing with such blood loss include allogeneic blood programs, preadmission donation programs, pharmacologic agents, hemodilution, and perioperative blood salvage. This article reviews these techniques as well as the consequences of perioperative anemia. Problems associated with blood transfusions are also outlined. PMID- 17131736 TI - Airport detection of implanted orthopedic devices. AB - Since the attacks of September 11, 2001, the United States and other countries have increased airport security. Metal detectors (archway and handheld wand) are used to detect potentially dangerous instruments during airport screening. One consequence is that orthopedic implants are sometimes detected, and patients are detained and searched. The orthopedic literature is inconsistent in its reports on airport screening of patients with orthopedic implants. In this article, I review that literature and examine the factors that make detection of orthopedic implants a less than consistent science. PMID- 17131737 TI - Postthrombotic syndrome after asymptomatic deep vein thrombosis following total knee and hip arthroplasty. AB - Lower extremity deep vein thrombosis (DVT) is relatively common after total knee arthroplasty (TKA) and total hip arthroplasty (THA). In this study, we sought to identify whether post-thrombotic syndrome is a significant risk in patients with asymptomatic DVT after TKA and THA. Included in the study were 186 patients (203 lower extremities) who had undergone routine lower extremity venography after TKA or THA between 1989 and 1993. Patients were contacted a minimum of 7 years after surgery. Postoperative varicose veins occurred in 8% of limbs without DVT compared with 12% of limbs with DVT. Hyperpigmentation occurred in 13% of limbs without DVT and in 16% and 18% of limbs with proximal and distal DVT, respectively. Swelling occurred in 6% of limbs without DVT, in 8% of limbs with proximal DVT, and in 12% of limbs with distal DVT. None of these differences was statistically significant, and there were no differences for proximal versus distal DVT. There was no significant increase in risk for postthrombotic syndrome after asymptomatic proximal or distal DVT after TKA or THA. PMID- 17131738 TI - Characteristics and incidence of fibromyalgia in patients who receive worker's compensation. AB - The aim of our study was to investigate the incidence of fibromyalgia syndrome (FMS) in patients with work-related injuries, the potential risk factors for and causes of FMS, and the disabilities associated with FMS. PMID- 17131739 TI - Arterial occlusion of vascular graft after total knee arthroplasty performed without tourniquet: a case report. PMID- 17131740 TI - Complete traumatic extrusion of the distal ulna through the hypothenar eminence: a case report. AB - We present a case of complete traumatic extrusion of the distal 40% of the ulna through the hypothenar eminence in the context of an open, both-bone forearm fracture and spinal cord transection. To the best of our knowledge, this specific trauma is unique to the literature, and, therefore, there is no standard treatment approach. Our patient was managed with irrigation and debridement along with open reduction and internal fixation of the radius fracture. No reconstruction of the ulna was performed. The patient reported excellent functional results at 13-month follow-up. PMID- 17131741 TI - Beyond mechanics. Advanced medicine for the moment of death. PMID- 17131742 TI - The pregnant MVA victim. An accident takes an ominous turn for a patient who's 38 weeks pregnant. PMID- 17131743 TI - Listening later: can some complaints really wait? PMID- 17131744 TI - Victim rescue drill: lessons learned. PMID- 17131745 TI - AHA guidelines say CPR is back in the spotlight ... for ZOLL, it never left. PMID- 17131746 TI - Setting the (tactical) scene. PMID- 17131747 TI - The tactical medic: not just for SWAT ops. PMID- 17131748 TI - Caring for the caregiver. PMID- 17131749 TI - Beyond the basics: obstetric emergencies. PMID- 17131750 TI - Paying it forward. The importance of being a mentor. PMID- 17131751 TI - Making the transition to filmless radiology. PMID- 17131752 TI - No single role for nurses in I.T. PMID- 17131753 TI - Is now the time for speech recognition? PMID- 17131754 TI - Readers' perspectives. Federal agencies have moved too slow to create policies that support President Bush's goal of electronic medical records. PMID- 17131755 TI - [The effect of the time of death on the reactivity of rat caudal artery regulated by Bay-K 8644 (an agonist directly affecting the Ca channel)]. AB - The research was conducted on Bay-K 8644-perfused rat caudal arteries in four different time groups. Investigations using the direct Ca channel agonist demonstrated a time-dependent decrease of arterial reactivity. The authors determined that the contractions of the smooth muscle coat preceded by an increased cytoplasm Ca++ level were significantly prolonged by the activation of direct Ca++ inflow from the extracellular space. PMID- 17131756 TI - [The effect of the time of death on the reactivity of rat caudal artery regulated by NA with a simultaneous use of NOS, CG and COX inhibitors. Part I]. AB - The investigations were carried out on NA-perfused rat caudal arteries in four different time groups. Inhibitors of NO synthase, CG and COX were used simultaneously to explain the effect of the time of death on biochemical mechanisms regulating the function of metabotropic receptors. The study demonstrated that using the NO synthase inhibitor improved post mortem arterial reactivity, what indicated maintained endogenous NO synthesis after death. The CG inhibitor was also found to improve arterial reactivity and intracellular Ca++ ion concentration. No improvement in arterial reactivity was observed when COX was used in the experiment. PMID- 17131758 TI - [Problems associated with chronological age estimation of children exploited in child pornography production]. AB - Chronological age assessment of young persons featuring in pornographic pictures and videos is crucial to prove a violation of law. The paper discusses possibilities of and difficulties inherent in age estimation in cases of production and distribution of child pornography. The presented problems were divided into technical and individual development-associated issues. Technical difficulties included lack of a reference system for biological features reconstruction, poor quality and resolution of pictures or movies, pictures retouching and photomontage. The author stressed that biological hindrances in age assessment were the consequences of interpersonal variation of developmental patterns, namely overlapping of ranges typical for feature values at particular ages and biological variations between different human populations. The described problems can render age estimation impossible or far from precise. Developing more accurate methods of estimating age from pictures and videos requires the collaboration of specialists in the field of auxology, anthropology, pediatrics, as well as experts in photography and video techniques. PMID- 17131757 TI - [The effect of the time of death on the reactivity of rat caudal artery regulated by PHE with a simultaneous use of NOS, CG and COX inhibitors. Part II]. AB - The investigations were conducted on PHE-perfused rat caudal arteries in four different time groups. Inhibitors of NO synthase, CG and COX were used simultaneously to explain the effect of the time of death on biochemical mechanisms controlling the functioning of metabotropic receptors. The authors demonstrated that using the NO synthase inhibitor improved post mortem arterial reactivity, what indicated maintained endogenous NO synthesis after death. The CG inhibitor also improved arterial reactivity and intracellular Ca++ ion concentration. No improvement was seen in arterial reactivity when COX was employed in the study. PMID- 17131760 TI - [Myocardial infarction as an occupational injury as represented in the materials collected at the Department of Forensic Medicine in Bydgoszcz in the years between 2000 and 2004]. AB - Myocardial infarction is defined as coagulational necrosis of the cardiac muscle caused by ischaemia. In the majority of cases, myocardial infarction is a consequence of atherosclerosis of the coronary arteries. Excessive physical or mental effort can lead to a sudden increase in the required supply of oxygen necessary for the proper functioning of the cardiac muscle and in consequence to cardiac muscle necrosis. The objective of the present study was to analyze cases of myocardial infarctions occurring at work opinionated at the Medico-Legal Institute in the years 2000-2004. The analysis included 12 civil court cases where the plaintiffs demanded compensation for falling victim to myocardial infarction in a work setting. In the majority of cases (11), it was concluded that professional tasks performed during work-time did not meet the criteria of an occupational accident and the most prominent factor that caused cardiac muscle necrosis was internal rather than external in character. In one case, where the plaintiff's professional duties were not only associated with mental stress and a prolonged lack of rest, but were also performed in adverse weather conditions and required sustained and considerable physical effort, it was established that working conditions exerted a significant effect on the occurrence of a myocardial infarction. Opinionating in the above presented cases posed a significant problem due to the lack of legal criteria that would define the term of an "occupational accident". PMID- 17131759 TI - [Correlations between haplogroup membership and Y-STR haplotype as a potential measure of quality control in forensic examinations]. AB - A correlation between particular Y-STR alleles from the so-called "minimal haplotype" and haplogroup membership of the Y chromosome was tested. We collected 146 Y chromosomes from haplogroups R1*, R1a1* and 1* and estimated the frequency of Y-STR alleles in each haplogroup. We then used different algorithms to assign a haplogroup to a haplotype, and tested their accuracy. Generally, a method based on calculation of haplotype similarity using the highest allele frequencies as modal values and assigning a score to each locus based on a ratio of allele frequencies turned out to give the most precise matches. However, using the same rules for Y chromosomes from other populations did not allow for precise estimation of their Y chromosome haplogroup frequencies. Possible explanations for this failure include interpopulation differences in haplotypes correlated with particular haplogroups, as well as a relatively small number of chromosomes analyzed. Potential uses for the presented method in forensics were also described. PMID- 17131761 TI - [Cases of homicide-suicide (post-aggression suicide) in the material of the Chair and Department of Forensic Medicine in Bydgoszcz]. AB - Two cases of homicide-suicide (post-aggression suicide) are presented in this report. In the first case, a young man had killed his wife and mother-in-law inflicting numerous stab wounds to the chest and neck of both victims, subsequently strangled his daughter with his bare hands and then committed suicide by cutting his veins. In the second instance, a woman had killed one of her sons and tried to murder another by giving both victims psychostimulants and medicines against worms, as well as stabbing them repeatedly. She then committed suicide by hanging. PMID- 17131762 TI - [Opinionating in cases of assessing the subject's ability to undergo imprisonment in materials collected by the Department of Forensic Medicine in Bydgoszcz in the years 1998-2003]. AB - Estimating the subject's ability to undergo imprisonment and determining whether there are any reasons for delaying or interrupting the execution of the penalty has become a serious problem. Currently, the Polish system lacks clear and uniform opinionating criteria, and the available sources provide only general guidelines. The present report outlines the legal criteria in force in such cases and describes the opinionating criteria used in the Department of Forensic Medicine in Bydgoszcz. The authors have also conducted an analysis of 87 opinions issued at the Department in the years 1998-2003 that evaluated the subject's ability to undergo imprisonment. The most frequent reasons for an individual seeking recognition as being unable to undergo imprisonment were as follows: internal diseases (41), neurological diseases (38), mental disorders (28), orthopedic problems (10) and other types of diseases in isolated cases. In nine instances, the subjects' health status was deemed to constitute a contraindication for imprisonment or detention. In 13 cases, in which the subject's files included an opinion previously formulated by a clinician and stating that the given individual was unable to undergo imprisonment, an analysis of the available medical records, as well as a thorough examination of the patient proved that there were no reasons that would contraindicate imprisonment. PMID- 17131763 TI - [Analysis of deaths caused by rail-vehicles in the materials collected by the Department of Forensic Medicine in Bydgoszcz in the years 1992-2002]. AB - The analysis focused on cases of death following accidents involving rail vehicles, in which autopsies were performed at the Department of Forensic Medicine in Bydgoszcz in the years 1992-2002. Sixty-five individuals died in train accidents (80%), 16 people (20%) were victims of tram accidents (20%). Males constituted 86% of the material, while females accounted for 14%. The highest number of rail-vehicle accidents was observed in 1992. The most common reason of death was multiorgan injury. In the subgroup of tram accidents, the authors noted no injuries of the highest severity, such as limb amputations, amputations or crushing of the head, or fragmentation or rupturing of the trunk. On the other hand, in railroad accidents, the percentage of the above injuries was 26%, 44% and 24.5%, respectively. The most severe injuries were noted in the group of people hit by a train while in prone position. PMID- 17131764 TI - [Determination of haloperidol in fingernail/toenails by LC-ESI-Ms]. AB - The report presents the possibility of using fingernails/ toenails to determine haloperidol levels. The described determinations were performed using the method of liquid chromatography coupled with electrospray-ionization mass spectrophotometry (LC-ESI-MS). In the course of the investigation, the authors developed a method for isolating haloperidol from nails and its identification. Determinations were performed in fingernail/toenail samples originating from individuals who had been administered haloperidol at least 6 months prior to sample collection. The materials demonstrated the presence of haloperidol in the following amount: fingernails - 67.3 +/- 6.49 pg/mg, toenails- 98.9 +/- 9.14 pg/mg. PMID- 17131765 TI - [Phylogeographic approach in the interpretation of mitochondrial DNA sequencing results in forensics]. AB - In recent years, forensic mitochondrial DNA analysis has been undertaken from an evolutionary perspective. In particular, the phylogeographic approach based on a phylogenetic analysis of the spatial distribution of mitochondrial haplotypes and haplogroups appears to be a useful tool in the interpretation of identification cases. In this study, the phylogeographic approach has been employed in the analysis of three difficult forensic cases, where single nucleotide, homoplasmic differences were found between the reference and evidentiary haplotypes. mtDNA sequence variation has been examined by the control region (HVS I and HVS II) direct sequencing. Additionally, in order to clarify the subhaplogroup status of the selected haplotypes, DNA sequences of entire mitochondrial genomes obtained from two samples representing J1b subclade have been analyzed. PMID- 17131766 TI - [Problems associated with medico-legal opinionating in spine injuries--a case report]. AB - The paper presents a case of a 44-year old male with a compression fracture of the thoracic spine. In view of incomplete medical records and divergent results of medical examinations, the case gave rise to considerable difficulties in medico-legal opinionating. PMID- 17131767 TI - [A rare case of suicide by a self-inflicted head injury]. AB - The report presents a rare case of suicide committed by the subject by dropping a tree trunk on his head using a suspended pulley system. The corpse of a 60-year old man was found in the cellar, his head encircled by a blood puddle. A razor was placed on the floor near his right hand. A large fragment of a tree trunk and a system of hooks and pulleys fastened to the ceiling and the floor were found at the place of the accident. The autopsy revealed extensive traumatic lesions of the skull and brain. PMID- 17131768 TI - Elderly patient assaults: empirical data from the Assaulted Staff Action Program (ASAP) with risk management implications for EMS personnel. AB - Some elderly patients may become violent in the course of receiving health care services. This paper reviews the largely cross-sectional research on elder assaultive patients in health care settings and patient assaults, including those by the elderly, in EMS services. Data on the assault characteristics of elderly patients are presented from a 15-year, retrospective, longitudinal study of the Assaulted Staff Action Program (ASAP). The findings and their implications for the delivery of health care services are discussed and a detailed series of risk management strategies to reduce assaults in EMS services are presented. PMID- 17131769 TI - Police trauma and cardiovascular disease: association between PTSD symptoms and metabolic syndrome. AB - Although prior evidence exists concerning the association between posttraumatic stress disorder (PTSD) and cardiovascular disease, few studies have examined associations of PTSD symptomatology and the metabolic syndrome in the high stress occupation of police work. The metabolic syndrome is a clustering of cardiovascular disease risk factors that have also been independently associated with psychological conditions. The aim of this study was to examine associations between the PTSD symptoms and metabolic syndrome in police officers. A stratified sample of 115 police officers was randomly selected from the Buffalo, NY Police Department. PTSD symptoms were measured with the Impact of Event scale (IES), divided into categories of subclinical, mild, moderate and severe symptom levels. The metabolic syndrome was considered present if three or more of its component parameters (obesity, elevated blood pressure, reduced high density lipoprotein (HDL) cholesterol, elevated triglycerides, and abnormal glucose levels) were present in each officer. Results indicated a significantly increased prevalence of the metabolic syndrome among those officers in the severe PTSD symptom category compared with the lowest PTSD severity category (prevalence ratio (PR) = 3.31, 95% C.I. = 1.19 - 9.22). Adjustment for age did not alter the association appreciably (PR = 3.12, 95% C.I. = 1.15 - 8.50). Adjustment for several demographic and lifestyle factors (age, education, smoking, alcohol intake) reduced the magnitude of the prevalence ratio slightly for the severe versus subclinical PTSD category (PR = 2.69, 95% C.I. = 0. 79 - 9.13), with adjustment for age and education accounting for most of the attenuation (PR = 2.71, 95% C.I. = 0.99 - 7.37). Thus, officers with severe PTSD symptoms were approximately three times more likely to have the metabolic syndrome and education may account for some of this association. PMID- 17131770 TI - Officer-involved shooting: reaction patterns, response protocols, and psychological intervention strategies. AB - Psychologists who work with law enforcement agencies may be called upon to respond to an officer-involved shooting (OIS). These need not be the most traumatic critical incidents in policing, but when they are, the reasons usually involve a mix of incident characteristics, officer response styles, and departmental handling. This article describes some of the psychological reactions experienced by officers during and following an OIS and provides a model of administrative, legal, mental health, and peer support services for officers in need. Finally, the article discusses several key roles that the police psychologist can play in the process of managing an OIS. PMID- 17131771 TI - Juvenile offender suicide: prevalence, risk factors, assessment, and crisis intervention protocols. AB - Suicide is the leading cause of death in juvenile detention and correctional facilities. Moreover incarcerated juvenile offenders have an estimated suicide risk four times greater than adolescents in the general population. To address the problem of juvenile offender suicide, this article describes the extent of the problem and protocols for intervening with this high-risk population. Procedures for crisis intervention are recommended, and we delineate and discuss the step-by-step sequential Roberts' 7 stage crisis assessment and crisis intervention model. The article concludes with an examination of the extent to which suicide assessment and prevention protocols are currently being implemented in juvenile justice facilities. The implementation of suicide prevention programs has been shown to decrease risk of suicide in juvenile offenders; however, much work is still needed to encourage facilities to implement comprehensive assessment and prevention programs nationwide. PMID- 17131772 TI - Incidence of male childhood sexual abuse and psychological sequelae in disaster workers exposed to a terrorist attack. AB - This study documents the prevalence of male childhood sexual abuse (CSA) and psychological sequelae in a sample of disaster workers deployed to the World Trade Center (WTC) site following the September 11, 2001 terrorist attack. There are limited data on male CSA and its psychological impact, especially on a large non-treatment seeking sample. As part of a mandatory medical screening program, workers were assessed with well-validated and widely used clinician interview and self-report measures following their involvement in the restoration of services to Ground Zero and surrounding areas of lower Manhattan. Frequency of CSA measured by the Traumatic Events Interview (TEI) was 4.3% (n = 92). Clinician interview and self-report data were analyzed using t-tests, revealing statistically significant relationships (but not clinically meaningful scores) between CSA and scores on the CAPS, PCL, BDI, STAXI, and SDS. Further analyses revealed that individuals endorsing CSA were three-times more likely to score high (vs. low) on the BDI and CAPS. Since disaster workers traditionally summon images of strength and mastery, professionals may overlook CSA and symptoms of depression and PTSD in this population. PMID- 17131773 TI - The impact of posttraumatic sress on Iraqi police. AB - This paper explores the psychological impact of posttraumatic stress on a sample of the Iraqi Police Service (IPS). Four separate surveys of lPS members were administered utilizing a 17-item National Center for PTSD checklist. In total, 231 of the 520 IPS (44%) achieved scores indicating that they had met the criteria for the diagnosis of PTSD. As a result, a two-hour lecture dealing with critical incidents, stress, cumulative stress, vicarious traumatization, posttraumatic stress disorder reactions to terrorism, compassion fatigue, burnout, and acute traumatic stress management (ATSM--which helps manage individual responses during traumatic events) was developed. This intervention was translated and conducted in Arabic for the IPS. It was noted that the majority of IPS surveyed never had been given information regarding traumatic stress symptoms or reactions, nor were they aware that they themselves might have traumatic stress. In addition, there is very little access to appropriate psychological assistance, either on the job or in the community. PMID- 17131774 TI - XDR-TB: entering the post-antibiotic era? PMID- 17131775 TI - PAL: a new and practical approach to lung health. PMID- 17131776 TI - False-positive tuberculin skin tests: what is the absolute effect of BCG and non tuberculous mycobacteria? AB - BACKGROUND: Despite certain drawbacks, the tuberculin skin test (TST) remains in widespread use. Important advantages of the TST are its low cost, simplicity and interpretation based on extensive published literature. However, TST specificity is reduced by bacille Calmette-Guerin (BCG) vaccination and exposure to non tuberculous mycobacteria (NTM). METHODS: To estimate TST specificity, we reviewed the published literature since 1966 regarding the effect of BCG vaccination and NTM infection on TST. Studies selected included healthy subjects with documented BCG vaccination status, including age at vaccination. Studies of NTM effect had used standardised NTM antigens in healthy subjects. RESULTS: In 24 studies involving 240,203 subjects BCG-vaccinated as infants, 20,406 (8.5%) had a TST of 10+ mm attributable to BCG, but only 56/5639 (1%) were TST-positive if tested > or =10 years after BCG. In 12 studies of 12,728 subjects vaccinated after their first birthday, 5314 (41.8%) had a false-positive TST of 10+ mm, and 191/898 (21.2%) after 10 years. Type of tuberculin test did not modify these results. In 18 studies involving 1,169,105 subjects, the absolute prevalence of false positive TST from NTM cross-reactivity ranged from 0.1% to 2.3% in different regions. CONCLUSIONS: The effect on TST of BCG received in infancy is minimal, especially > or =10 years after vaccination. BCG received after infancy produces more frequent, more persistent and larger TST reactions. NTM is not a clinically important cause of false-positive TST, except in populations with a high prevalence of NTM sensitisation and a very low prevalence of TB infection. PMID- 17131777 TI - Guidance for national tuberculosis programmes on the management of tuberculosis in children. Chapter 2: anti-tuberculosis treatment in children. AB - The management of children with TB should be in line with the Stop TB Strategy, taking into consideration the particular epidemiology and clinical presentation of TB in children. Obtaining good treatment outcomes depends on the application of standardised treatment regimens according to the relevant diagnostic category, with support for the child and carer that maximises adherence to treatment. A recent development in treatment recommendations is that, following a comprehensive literature review, ethambutol is now considered safe in children at a dose of 20 mg/kg (range 15-25 mg/kg) daily. Critical areas for further research include the optimal formulations and dosing of first- and second-line TB drugs and new drug development. PMID- 17131778 TI - Wilhelm Conrad Rotgen and the advent of thoracic radiology. PMID- 17131779 TI - Tuberculosis and tuberculosis control in European prisons. AB - OBJECTIVE: Correctional facilities have often been cited as reservoirs for tuberculosis (TB), presenting a potential threat to the general population. Although correctional facilities are recognised as ideal settings for interventions, little is known about the TB epidemiology within them. The purpose of our survey was to collect data on TB in prisons of the WHO European Region and on existing control measures. DESIGN: A questionnaire was sent to 52 EuroTB correspondents asking for 2002 data on the total number of inmates, number of prisoners with TB, resistance rates, screening strategies, monitoring and responsibilities. RESULTS: Twenty-two (42.3%) countries completed the questionnaire. The median TB notification rate was 232 per 100,000 inmates (0 17,808). Prisoners had up to 83.6 times more TB than civilians. The majority (90.9%) of the participating countries reported performing active screening for TB on entry into prison, with a median detection rate of 393/100,000 (42-2362). Of the respondent countries, 81.8% claimed to perform contact investigations and 86.4% to house infectious TB patients separately. CONCLUSION: Although response to this survey was only 42.3% and might be biased by a country's engagement in TB control in prisons, the results highlight the vulnerability of prisoners to TB and emphasise the need for adequate case-finding and containment strategies in prison. PMID- 17131780 TI - Risk factors for mortality among adult patients with newly diagnosed tuberculosis in Samara, Russia. AB - SETTING: Five out-patient tuberculosis (TB) clinics and one in-patient TB hospital in Samara, Russia. OBJECTIVE: To identify predictors of all-cause mortality among patients with newly diagnosed TB. DESIGN: A retrospective case control study of adult TB cases registered over a 5-year period from 1999 to 2003 using multivariate logistic regression modeling. Cases were defined as patients with TB who died within 12 months of diagnosis. Controls were patients with TB who survived at least 12 months. RESULTS: The case fatality rate was 3.6%. A total of 92 cases and 368 controls were identified. Mean age was 43 years; 71% were male; 4% were human immunodeficiency virus (HIV) seropositive. The median survival time for cases was 39.5 days. In multivariate analysis, independent predictors of mortality included bilateral lung involvement (OR 3.65), cavitary lesions on chest radiograph (CXR) (OR 2.45), symptoms >4 weeks at the time of diagnosis (OR 2.62), anemia (OR 5.24), and injection drug use (IDU) (OR 4.45), controlling for age. CONCLUSION: Advanced TB disease at the time of diagnosis (as demonstrated by having bilateral lung involvement, cavitary disease, symptoms >1 month, and anemia) and IDU were associated with increased TB mortality in Samara. Interventions targeted at improving earlier TB case diagnosis and treatment may help to reduce mortality among patients with tuberculosis in this region. PMID- 17131781 TI - Tuberculosis risks and socio-economic level: a case study of a city in the Brazilian south-east, 1998-2004. AB - OBJECTIVES: To explore tuberculosis (TB) risks in relation to potential determinants in the city of Sao Jose do Rio Preto, Sao Paulo State, Brazil; to analyse morbidity and mortality indicators in Sao Jose do Rio Preto, and to determine the relationship between the risk of TB and socio-economic level (SEL) using a geo-referenced information system (GIS) and the national census for 2000. METHOD: Standardised incidence rates and TB incidence and mortality rates were calculated. Socio-economic variables were determined using the statistical technique of principal component analysis. Data sources were the Sao Paulo State Data Analysis System (SEADE), the TB Notification Database (EPI-TB), the Information Department of the Brazilian Health Ministry (DATASUS), and the Brazilian Institute of Geography and Statistics (IBGE). New cases reported in 1998-1999 and 2003-2004 in the urban area of the city were geo-referenced and analysed. RESULTS: TB risk in the city is twice as high in areas of lower SEL than in areas with higher SEL. CONCLUSION: The identification of areas with different levels of risk enables the Municipal Health Department to propose innovative interventions to minimise the risk of disease at both individual and population level. PMID- 17131782 TI - Administrative monitoring of tuberculosis treatment in Switzerland. AB - SETTING: Treatment of tuberculosis (TB) is critically dependent on adherence. Directly observed treatment (DOT) has been shown to be effective. OBJECTIVE: To determine operational treatment outcome using administrative treatment monitoring (ATM) to assess the need for more vigorous promotion of DOT. DESIGN: Cohort study in eastern Switzerland, where ATM was started in 2002. Bi-monthly progress forms and a treatment outcome form (after 6 months) were sent to the treating doctors. Forms not returned within 6 weeks were followed up with phone calls. RESULTS: Between 2002 and 2004, 98 (87.5%) of 112 new TB patients completed a 6-month treatment course. Eight elderly patients died of causes other than TB while on treatment, four travelled out of the region and two were lost to follow-up. Treating doctors opted for DOT in only seven cases. CONCLUSION: Given the high success rate of 87.5% in our cohort, more vigorous promotion of DOT is not a priority for TB case management in eastern Switzerland. In our setting, ATM in collaboration with the family doctors offers a valuable alternative to the more time-consuming universal DOT. PMID- 17131783 TI - An Internet-based surveillance system for tuberculosis in Korea. AB - SETTING: The Korea Tuberculosis Surveillance (KTBS) network includes 248 health centres throughout the country, as well as other public and private health institutions. OBJECTIVE: To develop a web-based surveillance system for tuberculosis (TB) and to monitor implementation of the National TB Control Programme (NTP) on an ongoing basis. DESIGN: A TB notification form was developed with new case definitions, and standardised to obtain uniform essential information of the cases with ease and speed. Data collection, compilation, analysis and feedback were made available at every level of the health authority via the Internet without restrictions of time and space. RESULTS: The Internet based surveillance system was successfully implemented across the country, providing real-time national figures of TB using different variables-patient, time, area, site and type of disease--and facilitating on-line evaluation of NTP implementation. CONCLUSION: The web-based surveillance system has been well established within the existing health infrastructure, providing real-time figures on the TB burden. However, it requires continued improvement of the quality of information and of case reporting activities. PMID- 17131784 TI - Early and rapid microscopy-based diagnosis of true treatment failure and MDR-TB. AB - SETTING: Damien Foundation tuberculosis (TB) control project in Bangladesh. OBJECTIVE: Early diagnosis of true TB treatment failure and multidrug resistance (MDR) for more efficient DOTS-Plus. DESIGN: Prospective comparison of performance on smear-positive sputum of fluorescein diacetate (FDA) vital staining vs. culture, and of slide drug susceptibility testing (slide DST) vs. the Lowenstein Jensen (LJ) proportion method. RESULTS: FDA reached 92% positive and 97% negative predictive value directly on fresh sputum, but only 94% and 62%, respectively, on transported smears. Accuracy on washed cetylpyridinium chloride transported sputum was similar to that on fresh sputum. Slide DST on fresh smear-positive sputum failed less often than LJ DST, with 96% accurate results for rifampicin and MDR-TB diagnosis. Good results were obtained for isoniazid (90% accuracy), but not for ethambutol or streptomycin. CONCLUSIONS: We can confirm that FDA staining allows rapid screening for viable acid-fast bacilli and true treatment failure in delayed smear converters or smear-defined failures, while slide DST assures fast and accurate confirmation of MDR-TB in selected populations. The tests can be applied safely in resource-poor settings. Their successive use could be an efficient strategy for screening and an early start on standardised regimens of DOTS-Plus candidates. PMID- 17131785 TI - Extension of the intensive phase reduces unfavourable outcomes with the 8-month thioacetazone regimen. AB - SETTING: Damien Foundation tuberculosis (TB) control projects in Bangladesh. OBJECTIVE: To assess the effectiveness of a 1-month extension of the intensive phase for smear-positives at 2 months of an 8-month regimen with a continuation phase consisting of isoniazid (INH) and thioacetazone (Th). DESIGN: A prospective study of two cohorts of newly registered smear-positive cases, with extension of the intensive phase for the control cohort, but not for the study cohort. Culture and drug susceptibility testing (DST) of smear-defined failures and relapses and of random samples of new cases. RESULTS: Among 8230 study patients (86.7% 2-month conversion) and 7206 controls (83.4% conversion), smear-defined failure or relapse outcome was 3.0% for 2-month smear-negatives vs. 3.1% for 2-month smear positives with extension (non-significant, NS), and 8.2% for 2-month smear positives with no extension (P < 0.00001). Culture-confirmed failure and relapse reached 1.9% in 2-month smear-negatives and 1.6% (NS) in 2-month smear-positives with vs. 3.7% (P < 0.001) in 2-month smear-positives with no extension. The relative risk (RR) of non-extension in 2-month smear-positives was 2.4 (cultures) to 2.7 (smears). The same RR and borderline significance was found for non extension of patients with pan-susceptible strains. CONCLUSIONS: Extension of the intensive phase considerably reduces failures and relapses with a weaker regimen in patients smear-positive at 2 months. Its effectiveness may vary with extent of initial drug resistance vs. power of the regimen. PMID- 17131786 TI - Introduction of an in-house PCR for routine identification of M. tuberculosis in a low-income country. AB - SETTING: National Tuberculosis (TB) Treatment Centre, Makerere University Medical School and Joint Clinical Research Centre, Kampala, Uganda. OBJECTIVE: To evaluate the introduction of a polymerase chain reaction (PCR) based assay for identification of the Mycobacterium tuberculosis complex (MTC) into routine practice. DESIGN: Routine diagnostic specimens were processed and inoculated into Bactec 12B vials and monitored daily. At a growth index (GI) > or =10, 0.5 ml of the 12B broth was removed and assayed with PCR. The same 12B vial was analyzed using the Bactec NAP method at GI > or =500. Vials at various levels of GI were included. Recurrent cost and time required to perform PCR and NAP were compared. RESULTS: Initially, 71 specimens were analyzed; of these, 68 were NAP-positive while 69 were PCR-positive for MTC. PCR resulted in a 75% reduction in cost for a single test compared with Bactec NAP. PCR has been successfully incorporated into routine practice, and 432 samples have been analyzed. In addition, isolates from solid media were also well identified by PCR. With PCR, more samples can be analyzed at a time, it is faster and is less labor intensive. CONCLUSION: PCR is a reliable and cheaper alternative for the identification of MTC. PMID- 17131787 TI - Findings on the atopic triad from a Danish twin registry. AB - OBJECTIVE: To estimate to what extent the same genetic and environmental risk factors influence asthma, hay fever and eczema. DESIGN: From the nationwide Danish Twin Registry, twin cohorts born between 1953 and 1982 were contacted for a questionnaire survey, and a total of 29 183 twin individuals (86%) responded. Subjects were classified as cases when responding affirmatively to three questions about the lifetime occurrence of asthma, hay fever and eczema. Variance components twin analysis was conducted using maximum likelihood methods. RESULTS: The phenotypic (within-subject) correlations in liability between the different diseases were 0.57 (95% CI 0.54-0.59) for asthma and hay fever, 0.40 (95% CI 0.36 0.42) for asthma and eczema, and 0.33 (95% CI 0.29-0.36) for hay fever and eczema. Decomposition of these correlations into their genetic and environmental contributions showed that shared genes explained between 70% and 85% of the correlation between the different diseases. The remaining parts were explained by environmental factors shared between the diseases. CONCLUSION: To a large extent, atopic diseases share a common genetic background, although disease-specific genes also play a considerable role. These results can prove informative when counselling families with atopy, and may furthermore be used to guide the search for pleiotropic genes of importance for these diseases. PMID- 17131788 TI - Validation of the St. George's Respiratory Questionnaire in patients with COPD or asthma in Morocco. AB - SETTING: Chronic obstructive pulmonary disease (COPD) and asthma are a public health problem in Morocco. Their evaluation should be supplemented by quality of life measurement, but there is no specific instrument available in local Moroccan Arabic. OBJECTIVE: To validate a Moroccan Arabic version of the St George's Respiratory Questionnaire (SGRQ) in patients with COPD or asthma in Morocco. DESIGN: After a rigorous translation process of the SGRQ into local Moroccan Arabic (SGRQm), the SGRQm, spirometry, Fletcher scale and a visual analogical scale (VAS) were administered to each patient at recruitment. Data were used to examine the construct validity and reliability of the SGRQm. To examine the test retest reliability, patients completed the SGRQm a second time 10 days later. RESULTS: A total of 131 patients with a mean age of 52 years were recruited into the study. Patients had a mean forced expiratory volume in one second (FEV1) of 1.70 1 and a mean VAS of 51.5 mm. Internal consistency of symptoms, activity and impact components was assessed using Cronbach's alpha (a) reliability coefficient; they were 0.94, 0.91 and 0.90, respectively. The test-retest reliability of components scores ranged from 0.70 to 0.87. The Fletcher scale correlated with all SGRQm scores, while the VAS had a similar correlation except with symptoms (P > 0.05). FEV1 values were negatively correlated with all SGRQm scores. CONCLUSION: The SGRQm yielded satisfactory psychometric properties. PMID- 17131789 TI - Human immunodeficiency virus, smoking and self-rated health in Harare, Zimbabwe. AB - SETTING: Twenty-two urban factories in Harare. OBJECTIVE: To determine the relationship between the human immunodeficiency virus (HIV), smoking and self rated health in a high HIV prevalence urban workforce. DESIGN: Cross-sectional survey. RESULTS: Of 7482 employees, 6111 (82%) consented to interview and anonymous HIV serology; 88% were male; median age was 34 years. HIV prevalence was 19%. Current (median 6 cigarettes per day) and former smoking were reported by 17% and 7%, respectively. Smoking (current or former) was more common among HIV-positive (27%) than -negative participants (17%; P < 0.001). Factors significantly associated with being a smoker on multivariate analysis were being HIV-infected (OR 1.5, 95% CI 1.4-1.7), older age (P < 0.001), non-Christian (OR 1.6, 95% CI 1.2-2.2) and manual job (OR 1.4, 95% CI 1.2-1.6). Women (OR 0.05, 95% CI 0.03-0.11) and the better educated (OR 0.7, 95% CI 0.5-0.9) were significantly less likely to smoke. HIV-positive smokers had the highest risk of reporting poor health (adjusted OR compared to HIV-negative non-smokers 3.4, 95% CI 2.3-5.0). CONCLUSIONS: Smoking was significantly more common among HIV-positive than negative employees in this predominantly male workforce. There was evidence of a combined effect on self-rated poor health, a variable shown to be a strong independent predictor of mortality in industrialised countries. Interventions to encourage smoking cessation may be an important component of HIV care in Southern Africa. PMID- 17131790 TI - Pneumocystis jirovecii and microbiological findings in children with severe pneumonia in Nairobi, Kenya. AB - OBJECTIVE: To determine the significance of Pneumocystis jirovecii infection in the Kenyan paediatric population. DESIGN: Sixty samples of induced sputum from children aged < or =23 months, half of whom were human immunodeficiency virus (HIV) positive, admitted with severe pneumonia in Nairobi were subjected to immunofluorescent staining for detection of P. jirovecii and microbiological culture. RESULTS: P. jirovecii was detected in 8/60 (13%) as a copathogen with other respiratory pathogens. Five of eight samples with >5 oocysts were from HIV positive children aged < or =6 months, while equivocally scored samples (< or =5 oocysts) were from HIV-negative children aged >6 months. Klebsiella pneumoniae was significantly recovered in 26/ 60 (43%), followed by Escherichia coli 11/60 (18%) and Staphylococcus aureus 8/60 (13%). Streptococcus pneumoniae, Haemophilus influenzae and Pseudomonas aeruginosa were isolated infrequently. Candida albicans was recovered from 27/60 (45%), while the frequency of C. tropicalis, C. glabrata and C. parapsilosis was 7%, 5% and 3% respectively. Multidrug resistance among E. coli and K. pneumoniae were: sulphamethoxazoletrimethoprim 100% vs. 69%, chloramphenicol 55% vs. 73% and ampicillin 100% vs. 89%. CONCLUSION: Paediatricians in Kenya should be aware of Pneumocystis pneumonia, irrespective of the patient's HIV status. PMID- 17131791 TI - Tuberculosis control in Bolivia, Chile, Colombia and Peru: why does incidence vary so much between neighbors? AB - SETTING: In 2003, Peru and Bolivia reported the highest annual tuberculosis (TB) incidence rates in the Americas. Neighboring Colombia and Chile had lower annual incidence rates despite their proximity. OBJECTIVE: To determine what factors contribute to differences in TB incidence rates among Chile, Colombia, Bolivia and Peru. DESIGN: Multiple sources of literature dating between 1990 and 2005 were used and World Health Organization TB control guidelines were consulted for policy level comparisons. RESULTS: Comprehensive implementation of the DOTS strategy is the main factor explaining the differences in TB incidence rates, even after considering socio-economic factors. CONCLUSION: Cross-national comparisons suggest ways to improve regional DOTS implementation. PMID- 17131792 TI - Repeated sputum microscopy is not essential for monitoring tuberculosis treatment response. AB - SETTING: This study was conducted in two districts in India where DOTS has been implemented. There are 39 microscopy centres in Anantpur district and 34 in Nellore district (one per 100,000 population), each with a trained microscopist. Periodic follow-up sputum microscopy is performed for all tuberculosis (TB) patients on treatment, with two sputum specimens examined on each follow-up. Results are recorded in a laboratory register. OBJECTIVE: To determine whether examining two sputum specimens for follow-up is useful for assessing treatment outcome. DESIGN: A retrospective study using data from laboratory registers of all microscopy centres for 2002 in Anantpur and 2003 (January-June) in Nellore. RESULTS: Of 5086 follow-up examinations done in Anantpur and 1028 in Nellore, 8% were acid-fast bacilli positive. One additional positive result was obtained on examination of a second sputum specimen. This result did not significantly add to the assessment of treatment outcome. CONCLUSION: The yield of a second on-the spot sputum specimen is negligible, provided that the first smear is from an overnight specimen. From the data available, it is evident that repeated follow up smears are not essential for documenting treatment outcome. PMID- 17131793 TI - Contamination of stored sputum AFB smears with environmental mycobacteria. AB - To check the quality of stains and the acid-fast bacilli (AFB) staining procedure, a sample of 73 AFB-negative sputum smears selected systematically from a microscopy centre was rechecked before and after restaining with the same AFB staining method at a reference laboratory. Respectively 0 and 30 showed AFB before and after restaining. Detection of AFB after restaining in 30 of 73 negative smears triggered suspicion of contamination with environmental mycobacteria, which was confirmed by reexamination of these slides. Restaining before rechecking is a major limiting factor, and the results of external quality assessment should be ignored when contamination of slides is suspected. PMID- 17131794 TI - Saquinavir and rifampicin for tuberculosis and AIDS: new considerations. PMID- 17131795 TI - Multidrug-resistant tuberculosis in India. PMID- 17131796 TI - [Headache: diagnosis and treatment]. AB - Headache is the most common reason why patients visit a neurologist. Approximately 70% of the population suffer from headache. The symptoms in this condition are rather simple and a well-controlled therapy is mostly available, but there are still frequent failures in the diagnosis and therapy of headaches. Failures in the treatment are caused mainly by the fact the doctors are not able to set the appropriate diagnosis. It is recommended that patients with persisting headache should turn to headache centers. The therapy should be determined by the frequency and duration of the headache, its severity, the accompanying symptoms and preceding therapeutic history. This survey may be of assistance on the state of-the-art diagnostics and therapy of headaches in neurological practice. PMID- 17131797 TI - [25 years experiences on diabetic ketoacidotic patients]. AB - INTRODUCTION: In the past 25 years the authors regularly assessed the clinical characteristics of patients with diabetic ketoacidosis who were admitted to their diabetes unit. AIM: The aim was to examine the possible changes in the incidence, causes, treatment and mortality of diabetic metabolic disorders. METHODS: Retrospective analysis of the data obtained by the assessment of case histories. RESULTS: Between 1981 and 2005, 288 patients were admitted with 364 ketoacidotic episodes. The mean age of the patients was 42 +/-16.9 -- 53.2 +/- 12.6 years. Approximately 60% of them had type 1 diabetes and the proportion of the male patients was almost the same. The mean duration of diabetes was 7.8 +/- 8.1 -- 10 +/- 7.1 years. The ratio of manifest comas was approximately 10%. Among the precipitating factors the dominance of infections decreased (from 77.3% to 50.4%). During the treatment the aim of the authors was to supply fluids and electrolytes properly and to administer insulin for a prolonged period of time and in a dose as small as possible. The dose of the insulin bolus significantly decreased in the last period (8.29 +/- 4.31 U; p < 0.05). In the observed period 43 patients died (14.93% of the patients), the mortality rate decreased to 7.2% during the last period. CONCLUSIONS: Despite the intensive education, care and treatment of diabetic patients, the number of hospitalizations due to diabetic ketoacidosis has not decreased. In order to prevent ketoacidotic episodes proper team work is needed in the management of diabetic patients. PMID- 17131798 TI - [Radioiodine treatment of benign thyroid diseases]. AB - Radioiodine treatment of benign thyroid disease has a 60 year history. Among the radioactive isotopes (131)-I can be used successfully. This method has been spread at different rate over the world. Radioiodine is used in Hungary since 1959, however, incidence of this type of therapy does not reach an optimal proportion. The aim of therapy is to decrease the function and/or the volume of thyroid gland. This study considers the reasons of radioiodine treatment in Graves' disease and in different forms of autonomously functioning nodular thyroid disorders, moreover, in cases of compressive euthyroid multinodular goiter when the thyroid volume is to be diminished. There are controversies in terms of indications of radioiodine treatment, however, contraindications are generally well accepted. Sometimes radioiodine therapy could exert an unfavourable effect on ophthalmopathy but this can be prevented with steroids. In cases of suitable administration of thyrostatic treatment before (131)-I therapy the outcome of which does not change significantly though propylthiouracil has really some radioprotection effect. Excellent cure rate can be reached in Graves' disease with radioiodine but hypothyroidism after the treatment may be frequent. In cases of autonomously functioning nodular thyroid disorders (i.e., toxic uninodular or multinodular goiter) cure rate is as high as in Graves' disease but the incidence of thyroid hypofunction is much lower. Thyrotoxic crisis after the treatment is extremely rare. In some cases thyroiditis can be observed after (131)-I therapy but this condition can be easily managed. Thyroid radioiodine uptake and the effectiveness of (131)-I can be increased with the administration of recombinant human thyrotropin in cases of euthyroid multinodular goiter. Recently radioiodine is recommended to young patients having Graves' disease (over 15 years) in special cases. The risk of malignancy after isotope therapy is not significant. Radioiodine treatment of benign thyroid disorders is a simple, safe and cost-effective method, the propagation of which can be suggested. PMID- 17131799 TI - [Polymyalgia rheumatica]. AB - Polymyalgia rheumatica is a disorder that affects people over 50 years of age. The etiology of the disease has not been hitherto clarified exactly. Its incidence among people over 50 is in the range of 0.1-0.5%. The incidence rate peaks in the age group of 60-70 years. It is also found in younger people, but far less frequently. The diagnosis is based primarily on locomotor complains- namely on pronounced pain, morning stiffness of the shoulder girdle, pelvic girdle and neck. Complaints relating to the arms and legs (such as muscular weakness, oedema, tendonitis etc.) are also observed, however, in one third of the cases. The diagnostic criteria are defined empirically. Polymyalgia rheumatica was formerly considered to be a form of elderly onset rheumatoid arthritis. The progressive erosion process is absent in the case of polymyalgia rheumatica unlike in the case of rheumatoid arthritis. Numerous factors are known, which point to a link between polymyalgia rheumatica and giant cell vasculitis, arthritis, but the precise nature of this relationship remains unknown. Both conditions affect the same age group in the general population and they are even found--not infrequently--in the same patient. Polymyalgia rheumatica can be found in 40% of the patients suffering from arthritis while the histological examination detected mild vasculitis in approximately 10% of the patients suffering for "isolated" polymyalgia rheumatica. The response to be given to the acute phase is similar in both disorders. Scandinavian authors consider polymyalgia rheumatica as the appearance of generalised arthritis. Arthroscopic, nuclear magnetic resonance imaging as well as isotopic studies show unequivocally, that in the background of the osteo-muscular symptoms, complaints, inflammation is to be found partly of the joints but primarily that of the periarticular synovial structures. The above mentioned--dominant--proximal symptoms can often mask the distal locomotor disorders (pitting oedema of the hands and feet, tendonitis, tendosynovitis, carpal tunnel syndrome). The disorder may be accompanied by atypical generalised symptoms (loss of appetite, weight loss, fever, fatigue). An excellent indicators of the acute phase reactions are erythrocyte sedimentation rate, C-reactive protein and interleukin-6. These are suitable for monitoring the effectiveness of the therapy, for indicating a relapse/recurrence. It should be noted, that polymyalgia rheumatica may also be present if the erythrocyte sedimentation rate and C-reactive protein values are low. This disorder is also characterised by fast and effective response to corticosteroid, which should be administered for 1-2 years. In some individual cases a different dosage regime may be necessary: steroid administered in low dosage over a longer period of time. Administration of methotrexate and anti tumor necrotic factor-alpha may also be considered as alternative or adjuvant therapy for lowering the quantity of corticosteroid. Further multicenter, double blind studies should, however, be performed on large number of patients in this regard. PMID- 17131800 TI - [Incomplete androgen insensitivity]. AB - In the androgen insensitivity syndrome (AIS) the androgen effect is decreased in the fetus and the youth despite the adequate testosterone production. Usually the mutation of the androgen receptor is responsible for the disease. In the presented case the external genitalia were similar to female genitalia but masses were palpable in the labioscrotal fold. The karyotype was 46,XY. There was no increase in the testosterone level during the first three months of life. The stimulation test by stanazolol and the androgen receptor gene analysis verified the androgen insensitivity. The mutation was absent in the mother's leukocytes. This fact makes the genetic advising difficult in this family. PMID- 17131801 TI - A universal concept. PMID- 17131802 TI - A loving challenge. PMID- 17131803 TI - A shift of balance. PMID- 17131804 TI - Therapeutic electricity. PMID- 17131805 TI - Empowering mobility. PMID- 17131806 TI - Stimulating therapy. PMID- 17131807 TI - [Hi-tech trends in future dentistry--Part B: diagnostic and therapeutic technologies]. AB - The technology advance and the growing amount of knowledge had a great impact on dental practice for the last decade. The prominent change began with digital revolution presenting new computed technologies and accessible communicational means for sharing literal and imaging information. Following toward the coming biotechnological revolution, dentistry will be even further changed. This article presents dental innovations of Israeli Hi-Tech companies, sorted into two groups. Part A of the article discussed the area of computed imaging systems for educational purposes, diagnostic and treatment. While part B presents other diagnostic or therapeutic technologies. However, because some of the described technologies are still on their R&D (Research and Development) phases, they are not commercialized yet in the market. PMID- 17131808 TI - [Intra-oral digital photography with the non professional camera--simplicity and effectiveness at a low price]. AB - Over the last five years digital photography has become ubiquitous. For the family photo album, a 4 or 5 megapixel camera costing about 2000 NIS will produce satisfactory results for most people. However, for intra-oral photography the common wisdom holds that only professional photographic equipment is up to the task. Such equipment typically costs around 12,000 NIS and includes the camera body, an attachable macro lens and a ringflash. The following article challenges this conception. Although professional equipment does produce the most exemplary results, a highly effective database of clinical pictures can be compiled even with a "non-professional" digital camera. Since the year 2002, my clinical work has been routinely documented with digital cameras of the Nikon CoolPix series. The advantages are that these digicams are economical both in price and in size and allow easy transport and operation when compared to their expensive and bulky professional counterparts. The details of how to use a non-professional digicam to produce and maintain an effective clinical picture database, for documentation, monitoring, demonstration and professional fulfillment, are described below. PMID- 17131809 TI - [Gerodontology teaching program at the geriatric dental clinic in Yad Sarah]. AB - OBJECTIVES: World demographic changes show an increase in the elderly population worldwide. Due to increase in life span, the continuously growing elderly population requires extra attention. This has a great effect on private medical practice. A long term result is the necessity of training dentists in the practice of geriatric dentistry. Geriatric patients have problems that are age dependent, general health problems and specific dental problems. These problems require special skills on the part of the dentist. Yad Sarah has undertaken responsibility for this important task and provides dentists with a geriatric dental (gerodontology) training program. Elderly patients in need of geriatric dental treatment are served in both stationary and mobile clinics. The program includes both theoretical and practical studies. For this purpose, the Hebrew University-Hadassah School of Dental Medicine and Yad Sarah have created a three year program at the end of which the dentists, working voluntarily, receive a diploma in geriatric dentistry. The purpose of the present study is to evaluate the teaching program in the past few years. METHODS: All the lectures given in the program from 2001 to 2005, both theoretical and practical, have been summarized and categorized by subject. RESULTS: In 2001-2002, the teaching program was theoretical only. Starting from 2003 practical studies were added to the program so that now it includes both theoretical and practical studies. The results presented do not include the many clinical teaching hours during supervised treatment that are given privately to the dentists, from the preparation of the dental treatment plan until the conclusion of treatment. The result shows a steady increase in teaching hours during the years 2001-2005. Not only has the quantity increased; the content has changed too, because of the changes in the needs and the character of work in the geriatric dentistry field. The increasing activity through the mobile dental clinic for homebound patients has led to stepped up teaching in subjects that are related to this kind of treatment, such as oral medicine, systemic disease and oral rehabilitation relating to at-home treatment. The increase in implant treatments created the need to train the dentists in the surgical and rehabilitative aspects of care. The area of oral medicine (26%) and oral rehabilitation (24%) received the most teaching hours, while endodontics (4%) received the least. CONCLUSIONS: The teaching program at Yad Sarah is need-dependent--needs of the patients and of the volunteer dentists as well. At Yad Sarah the dentists come to volunteer, to give needy people quality dental treatment that they cannot afford themselves at regular clinics. An additional motive of these dentists is to advance themselves professionally. The Yad Sarah Dental Clinic offers a unique teaching program that contributes to enhancing knowledge in gerodontology. As the needs at the clinic change, so does the teaching program. PMID- 17131810 TI - Water fluoridation, again... PMID- 17131811 TI - [Urocortin decreases phosphorylation of MYPT1 and increases the myosin phosphatase activity via elevation of the intracellular level of cAMP]. AB - Urocortin, a peptide hormone related to the corticotropin releasing factor, is suggested to be involved in blood pressure regulation by dilating the peripheral blood vessels. In rat tail arteries, urocortin-induced vasodilation is due to a decrease in myofilament Ca2+ sensitivity the mechanism of which is still unclear. In this study, the hypothesis was tested that the decrease in Ca2+ sensitivity in mouse tail arteries results from the activation of myosin light chain phosphatase. The relaxation of KCl-precontracted (42 mM) intact mouse tail arteries by urocortin (1 nM and 10 nM) was significantly inhibited by 1 microM antisauvagine30, a CRF-2 receptor antagonist (p < 0.05, n = 3). The addition of 1 microM KT 5720, an inhibitor of PKA, to intact rat tail arteries did not affect the KCl-induced force but significantly attenuated the urocortin-induced relaxation (n = 5). In alpha-toxin permeabilized mouse tail arteries, urocortin relaxed submaximally activated preparations at constant pCa 6.1 by 37.6 +/- 8.2% (n = 5) as compared to control vessels (n = 5, p < 0.001). The relaxation in permeabilized vessels was inhibited by pre-treatment with 30 microM Rp-8-CPT cAMPS, an inactive analogue of cAMP. In permeabilized mouse tail arteries, treatment with 100 nM urocortin was associated with dephosphorylation of MLC20(Ser19) and MYPT1(Thr696/Thr850). The effect of urocortin on MYPTI dephosphorylation was completely abolished by 30 M Rp-8-CPT-cAMPS and mimicked by the cAMP analogue Sp-5,6-DCI-cBiMPS. Based on these findings, we propose that the urocortin-induced relaxation is due to a decrease in calcium sensitivity mediated by a cAMP-dependent increase in the activity of MLCP. PMID- 17131812 TI - [The in vitro motility assay to study the calcium-mechanical relationship in skeletal and cardiac muscles]. AB - In a series of experiments on regulated contractile systems (i.e., in vitro mobile systems with reconstructed thin filaments), the velocities of the movement of a thin filament on the surface covered by either rabbit skeletal or rat cardiac myosin at various concentrations of calcium ions in solution (in the pCa range from 4 to 8) were assessed. The corresponding "pCa-velocity" relationships were plotted, which proved to be of the sigmoid form. It was found that, at a saturating calcium concentration (pCa 4), the velocity of regulated thin filaments was 65% higher than for unregulated ones in the case of skeletal myosin and 87% higher than for unregulated thin filaments in the case of cardiac myosin. It was also found that the Hill coefficient was 1.95 and 2.5 for skeletal and cardiac myosins, respectively. The difference in the Hill coefficients for skeletal and cardiac myosins is discussed in terms of the difference in contribution of cooperativity mechanisms of contractile and regulatory proteins in the regulation of contraction in these types of muscles. PMID- 17131813 TI - [Thin filament elasticity and its role in the muscle contraction]. AB - The available experimental methods do not allow one to establish unambiguously the molecular structural events during muscle contraction. To resolve the existing controversies, I have devised an unconventional original computer program. The new approach allows the reconstruction of the hexagonal lattice of the sarcomere for different muscle states and verification of the structure by comparison of the calculated Fourier spectra with the real diffraction patterns. Previously, by the use of this approach, the real structure of a myosin filament from vertebrate striated muscle has been reconstructed (http://zope.ibib.waw.pl/pspk). In this work, a reconstruction for the thin filament is presented for three states: relaxed, after activation, and during contraction. Good consistency of the calculated Fourier spectra with the real diffraction patterns available in the literature suggests that the thin filament, due to flexibility, plays an active part in muscle contraction, as myosin cross bridges do. PMID- 17131814 TI - [Electron microscopic study of the influence of fullerene on the formation of amyloid fibrils by A beta(25-35) peptide]. AB - The anti-amyloidogenic capacity of hydrated fullerene C60 HyFn was revealed by the use of electron microscopy. We first showed that when, connecting with growing amyloid fibrils formed by A beta(25-35)-peptide, fullerene prevented their subsequent growth and interfered with the formation of new fibrils. Instead of long helically twisted ribbons formed by A beta(25-35)-peptide in the absence of fullerene, short narrow protofibrils were found in the presence of fullerene . These results allow one to suppose that fullerene can be useful for the therapy of Alzheimer's disease. PMID- 17131815 TI - [Cytotoxicity of amyloid fibrils of X-protein]. AB - It is known that amyloid oligomers, protofibrils, and fibrils induce cell death, and antibiotic tetracycline inhibits the fibrillization of beta amyloid peptides and other amyloidogenic proteins and disassembles their pre-formed fibrils. Earlier we have demonstrated that sarcomeric cytoskeletal proteins of the titin family (X-, C-, and H-proteins) are capable to form in vitro amyloid fibrils, and tetracycline effectively destroys these fibrils. Here we show that the viability of polymorphonuclear leukocytes in the presence of X-protein amyloids depends on the concentration of amyloid fibrils of X-protein and the time of incubation. In addition to the disaggregation of X-protein fibrils, tetracycline eliminated the cytotoxic effect of the protein. The antibiotic itself did not show a toxic effect, and the cell viability in its presence even increased. Our results evidence the potential of this approach for evaluating the effectiveness of drugs preventing or treating amyloidoses. PMID- 17131816 TI - [The C-terminal fragment of thymopoietin forms F-actin bundles: electron microscopic data]. AB - It was shown by electron microscopy that PEP33 a synthetic C-terminal peptide of the thymus hormone thymopoietin, formed bundles of actin 'filaments in the presence of 0.1 M KCl. The structure of PEP33 aggregates localizated in the bundles between actin filaments is very similar to that of aggregates observed in samples of pure PEP33. No changes were revealed in the structure of G-actin in the presence of PEP33. A similar, but a weaker bundling effect of thymopentin (PEP5) was also found. It forms bundles of actin filaments of small size. Further studies can shed light on the physiological importance of actin filament aggregation with the peptides of thymopoietin, the systematic release of which from the thymus produces the phenomena characteristic for the serious neuromuscular disease myasthenia gravis. PMID- 17131817 TI - [Thermostable extracellular cyclic nucleotide phosphodiesterase from Physarum polycephalum plasmodium]. AB - The cyclic nucleotide phosphodiesterase secreted by Physarum polycephalum plasmodium into extracellular medium has been partially purified by DEAE cellulose chromatography, ultrafiltration, and HPLC. The results obtained by gel filtration, HPLC, electrophoresis, and isoelectric focusing suggest that, the native enzyme in solution is a monomer with a molecular mass of about 90 kDa and pI in the range 3.6 - 4.0. The Km values were estimated to be about 0.9 mM and 7.7 mM, respectively, and Vm for both substrates were similar (up to several thousand micromoles of cAMP hydrolyzed/hour per mg of enzyme). The partially purified enzyme was shown to be extremely stable. It did not lose the activity after heat treatment at 100 degrees C during 30 min. The enzyme was active in the presence of 1% SDS, but it was fully inactivated under the same conditions in the presence of beta-mercaptoethanol. The properties of the phosphodiesterase from Physarum polycephalum are discussed. PMID- 17131818 TI - [Calcium-dependent interaction of transducin with calmodulin-sepharose]. AB - It has been shown by affinity chromatography on calmodulin-sepharose that transducin, a G protein of bovine retinal rod outer segments interacts with Ca(2+)-calmodulin. This result assumes that the main part of calmodulin in dark retinal rod outer segments is associated with transducin. It has been suggested that photoactivation of retinal rods induces changes in intracellular calmodulin concentration, which may be one of the steps involved in the light adaptation of photoreceptor. PMID- 17131819 TI - [Solubility and phase transitions in the water-protein-salt system]. AB - The previously derived formulas for the curves corresponding to sol-gel, liquid liquid, and liquid-solid phase transitions, which correlate the critical molar composition of the water-protein-salt system with individual characteristic features of its component (protein charge z, the number of ions adsorbed v, the function of electrolyte activity A) are presented as curves in ordinary coordinates of protein solubility logS against salt concentration m3. Tendencies in changes in phase transition lines versus the v, z, and v/z ratio have been determined. Correlations of the salting-out curve and the salting-out coefficient with phase transitions are discussed. PMID- 17131820 TI - [The influence of hindlimb unloading on the effectiveness of modulation of the mediator secretion via the autoreceptor system]. AB - In experiments on neuromuscular synapses of rat fast (m. Extensor digitorum longus, EDL) and slow (m. soleus) skeletal muscles, changes in the intensity of spontaneous quantal mediator secretion in response to the activation of presynaptic cholinoreceptors by the nonhydrolyzable acetylcholine analogue carbachol and to an increase in K+ concentration in the control group of animals and in animals subjected to different terms of unloading of hindlimbs have been compared. The intensity of spontaneous secretion of mediator quanta was evaluated from the mean frequency of miniature endplate potentials. In the control group of animals, the frequency of miniature endplate potentials by the action of carbachol increased by 363% in m. EDL and by 62% in m. soleus. The frequency of miniature endplate potentials in the synapses of m. EDL was more sensitive to K(+)-induced depolarization too. The bearing unloading of hindlimbs abolished the sensitivity of spontaneous secretion to carbachol in the synapses of m. EDL, whereas in m. soleus it was unchanged. However, the preservation of sensitivity of nerve endings of fast muscle to K(+)-induced depolarization allows one to assume that the hindlimb unloading leads to a decrease in the number of functioning presynaptic receptors. PMID- 17131821 TI - [Reduction of erythrocyte deformability in Krushinskii-Molodkina rats in acute hemorrhagic cerebrovascular circulation disorders]. AB - The ability of erythrocytes to change their shapes in the shear flow under acute strokes of hemorrhagic type in rats of the Krushinsky-Molodkina line was studied. The rigidity of membranes and the internal viscosity of erythrocytes were investigated by the laser diffraction method. The method consists in obtaining diffraction images from a thin layer of a dilute suspension of erythrocytes moving in the shear flow and subsequent computer processing of these images. It was shown that strokes of hemorrhagical type in rats of the Krushinsky-Molodkina line cause a reduction in the ability of erythrocytes to change theirs shapes. PMID- 17131822 TI - [Expression of presenilin 1 on the cell surface in motile polarized cells]. AB - Most cases of familial early-onset Alzheimer's disease are caused by mutations in the presenilin 1 (PS1) gene. However, the cellular functions of PS1 are not yet completely understood. We showed that endogenous PS1 and the adhesion protein CD44 are redistributed on the surface of cell projections (lamellipodia) in polarized T- lymphocytes (Jurkat cells) after the adhesion to a collagen matrix. This effect was not observed for another surface protein of T lymphocytes, which is not involved in cell adhesion processes, the T cell receptor. In primary cultures of mouse cortical neurons, PS1 was concentrated at the surface of extended growth cones and at the sites of neurite contacts. The concentration of PS1 at the surface of cellular structures that promote cell motility and cell contacts suggests an important role of PSI in cell adhesion in motile polarized cells. PMID- 17131823 TI - [Measuring real-time sarcoplasmic reticulum calcium pumping in skinned muscle fibers]. AB - A method and a device for direct measurements of accumulation of calcium in the sarcoplasmic reticulum (SR) and its release from SR as a function of free Ca2+ in bath have been developed. About 30% of the volume inside muscle fibers of swimbladder of Opsanus tau is occupied by SR. A set of solutions was prepared for fiber dissection and making holes in outer membrane without destruction of membranes of the sarcoplasmic reticulum. Calcium was unloaded from SR using EGTA as a pCa buffer. Then solutions with 50-100 microM CaFURA2 or CabisFURA2 were used as pCa-buffer and fluorescent Ca-indicators for measurement of Ca exchange between a fiber with a volume of approximately 10 nl and a solution in the cuvette with a volume of 5 microl. An increase in fluorescence signified an increase in unbound FURA in the bath since Ca2+ pumped into the SR was removed from the bath. The slope represented the rate of Ca2+ uptake by the SR in the muscle fiber, the maximum being about 1.6 M/s per liter of solution in bath or 2.6 mM/s per liter of SR volume. In solutions without oxalate and Ruthenium Red, more Ca2+ was taken up by the SR, and oscillations of the bath free FURA level were often observed, which can be explained by calcium-induced calcium release. PMID- 17131824 TI - [Cytoplasmic K/Na balance in heart muscle cells of young and old rats under oxygen and substrate deficiency]. AB - Age-related changes in the content of the major cellular cations of potassium and sodium in heart muscle cells of Wistar rats have been studied. The cytoplasmic concentration of potassium and sodium was determined by the electron probe microanalysis. The results revealed differences in both the concentration of the elements in young and old control animals and the responses of a cardiomyocyte to the state of acute hypoxic deenergization modelled on a perfused heart. The data are consistent with the hypothesis about the presence of genetically-related age changes in the conductance of potassium channels, which in old animals are realized against the background of deficient supply of tissues with oxygen and substrate. PMID- 17131825 TI - [Chemotaxis as a method for testing of the biological effects of silver nanoparticles]. AB - A method for assessing the abiotic efficiency of water-dispersed nano-sized silver particles is suggested. Nanoparticles were obtained by the method of biochemical synthesis in reversed micelles made from anionic surfactant bis-(2 ethylhexyl) sodium sulfosuccinate (AOT). A water dispersion of nanoparticles was prepared from the micellar solution in organic solvent by means of a special procedure. The abiotic efficiency assay is based on the capacity of cells to respond by the negative chemotaxis to chemical agents with harmful metabolic action. In plasmodium of myxomicete Physarum polycephalum, the biocide and repellent effects of silver nanoparticles, Ag+ ions, and AOT were tested in order to compare the abiotic efficiency of these substances in water solution and after introduction into agar substrate. The increase in the oscillation period, the reduction in the area of spreading, and the avoidance reaction in the spatial test, taken together, revealed a much higher repellent efficiency of silver nanoparticles as compared with that of Ag+ ions and AOT. The NSP concentrations lethal for Physarum were similar to those found earlier for bacteria and viruses. The chemotaxis-based tests applied in this study allow one to quantitatively assess cell reactions and monitor their time course. Besides, they have a much higher resolving capacity than the tests based on lethal effects of abiotic substances. PMID- 17131826 TI - [KRP/telokin differentially regulates filament assembly and phosphorylation of light chains of non-muscle myosin II in fibroblasts]. AB - Transgenic 3T3 fibroblasts have been generated that express either the wild-type KRP or its truncated mutant lacking the C-terminal domain, which primarily contributes to myosin binding of KRP. It was found that KRP-expressing cells display a significantly increased content of myosin filaments and a reduced level of rMLC phosphorylation, whereas the mock transfected cells or cells expressing the C-terminally truncated KRP do not. Our results suggest that (1) KRP promotes the polymerization of myosin II and reduces the rMLC phosphorylation level in cells, (2) KRP acts through direct binding to myosin II, and (3) transgenic 3T3 fibroblasts stably expressing KRP represent a useful and versatile model to study the role of myosin II filament dynamics in cell motility. PMID- 17131827 TI - [Antimicrotubule agents can activate different apoptotic pathways]. AB - The effect of agents (taxol, vincristine, and nocodazole) disturbing the microtubule network in MCF-7 human breast carcinoma cells has been examined. The aim of the study was to determine the subtypes of mitotic catastrophe and the dependence of cell death on the status of protein p53. Antimicrotubule agents can not only induce mitotic catastrophe, that is, cell death during mitosis and the death of micronucleated cells, but also activate apoptosis in interphase cells. We assume that the G1 checkpoint activation in this case occurs as a result of microtubule disruption. Apoptosis can be activated in a p53-independent manner in K-mitotic cells and after the complete disruption of the microtubule network. PMID- 17131828 TI - [Stochastic computer model of cellular microtubule dynamics]. AB - A computer model of the system of microtubules has been developed to study the mechanisms of action of various factors on this system. The model describes the process of polymerization/depolymerization of microtubules as a set of chemical reactions with certain rate constants using a stochastic approach. Microtubules are visualized in the program field, which makes the model visual. The program imitates the dynamics and structure of the system of cellular microtubules with great, reliability. The parameters generated by the model correlate with the corresponding parameters of microtubules in living cells. We are going to develop this approach to modeling microtubules and similar structures to bring them into a better accord with living systems and to study the influence of various factors on these systems. PMID- 17131829 TI - [Dynamic instabilities in microtubule cytoskeleton. A phase diagram]. AB - Instabilities in the growth and depolymerization of microtubules are considered in the framework of self-organization theory. An extended reaction-diffusion model for the microtubule dynamics has been formulated. A phase diagram of microtubule cytoskeleton has been constructed, which determines the regions of stability for steady and nonstationary solutions of the model. It is shown that the instabilities in microtubule dynamics result from kinetic nonequilibrium phase transitions. On the basis of phase diagram structure, a general classification of the microtubule cytostatic regulatory factors is suggested. The problem of mutual amplification of the activity of cytostatic agents is discussed. PMID- 17131830 TI - [Phalloidin suppresses the force in nebulin-rich lamprey cardiac muscle]. AB - The effect of phalloidin, an agent detaching nebulin from actin in skeletal muscle, on the isometric force in lamprey skinned cardiac muscle, which has nebulin in amounts comparable to that in skeletal muscle, has been studied. We found that, unlike mammalian cardiac muscle expressing nebulin less abundantly and responding to phalloidin by a force increase, lamprey cardiac muscle responds to phalloidin by a force decrease (approximately 50% decrease), thereby resembling the response of skeletal muscle. These results support our hypothesis that nebulin detachment from actin underlies phalloidin-induced force loss and suggest a role of actin-nebulin interaction in contractile function. PMID- 17131831 TI - [Effects of agonists and antagonists of rhyanodine receptors on potassium contractures in twitch and tonic frog skeletal muscle fibers]. AB - A comparative analysis of the effects of the concentrations of Ca2+ in external medium and the inhibitor (dantrolene) and activator (4-chloro-m-cresol) of rhyanodine-sensitive Ca2+ channels of carcoplasmic reticulum on the characteristics of potassium contracture in frog twitch and tonic skeletal muscles has been performed. It was shown that the duration of contracture in tonic muscles is not restricted by the presence of Ca2+, as distinct from twitch muscles. Dandrolene does not practically affect the contractile responses of tonic fibres, and the concentration of cresol eliciting the contracture for tonic fibres is substantially higher (1 mM) than for twitch fibers (0.25 mM). In twitch fibers, the potassium contracture activated in the presence of cresol is comparable in amplitude and dynamics with the contracture under control conditions, and in tonic fibers a summing of responses without relaxation after the washing of excessive potassium is observed. This suggests that, in twitch fibers, the influx of Ca2+ can directly create the concentration sufficient for the maintenance of contraction, and in tonic fibers its involvement is mediated through the Ca(2+)-dependent activation of the beta-isoform of rhyanodine sensitive channels. PMID- 17131832 TI - [Role of the alpha2-isoform of Na+, K(+)-ATPase in the positive inotropic effect of ouabain and marinobufagenin in the rat diaphragm]. AB - It was shown that the specific inhibitors of Na+, K(+)-ATPase ouabain and marinobufagenin increased the contraction of an isolated rat diaphragm (positive inotropic effect) by up to approximately 15% in a dose-dependent manner with EC50 = 1.2 +/- 0.3 and 0.3 +/- 0.1 nM, respectively. The results indicate the involvement of the ouabain-sensitive alpha 2 isoform of Na+, K(+)-ATPase. The analysis of ouabain-resting membrane potential dose-response relationships in the presence and absence of hyperpolarizing concentration of acetylcholine (100 nM) suggests the existence of two pools of alpha 2 Na+, K(+)-ATPase with different affinities for ouabain. The pool with a higher ouabain affinity is involved in the hyperpolarizing effect of acetylcholine and, presumably, in the positive inotropic effect of ouabain, which might be a mechanism of regulation of muscle efficiency by circulating endogenous inhibitors of Na+, K(+)-ATPase. PMID- 17131833 TI - [Elevation of the vascular smooth muscle sensitivity to effects of constrictors after denervation and under decreased blood pressure]. AB - Changes in contractile activity of saphenous artery in normotensive rats and in rats with regional hypotension have been investigated. The abdominal aorta was partially occluded in Wistar rats distally to the renal arteries. Four weeks later, a 5-7-mm segment of the femoral nerve in one hindlimb was resected to denervate the saphenous artery. After two weeks, the isometric contraction of innervated and denervated saphenous artery segments was studied. In normotensive rats, the denervation augmented vessel sensitivity to noradrenaline, phenylephrine, serotonin, and KCl (in the presence of phentolamine). Chronic hypotension also augmented vessel sensitivity to constrictor agonists, whereas denervation did not result in further increase of sensitivity. In glyoxilic acid stained preparations obtained from hypotensive rats, a reduced intensity of fluorescence of adrenergic fibers was observed. It was assumed that the higher sensitivity of vascular smooth muscle in hypotensive rats is due to functional disturbances of sympathetic innervation. PMID- 17131834 TI - [NFATc1 and slow-to-fast shift of myosin heavy chain isoforms under functional unloading of the rat m. soleus]. AB - It was determined whether the content of NFATc1 (nuclear factor of activation of T cells) in the nuclear and cytoplasmic extracts is related to an increase in the content of fibers containing type IIa myosin heavy chains under gravitational unloading of m. soleus. It was found that three isoforms of NFATc1 with molecular masses of 140, 110, and 86 kDa are present in m. soleus. Under unloading, the translocation of 140 kDa NFATc1 into the nucleus, a decrease in the content of 110 kDa NFATc1 in the cytoplasmic extract of m. soleus, and an increase in the content of 86 kDa NFATc1 in the nuclear extract of m. soleus take place. The content of fibers containing type IIa myosin heavy chains under gravitational unloading increases. The increase in the level of 140 and 86 kDa NFATc1 in the nucleus is accompanied by a decrease in the percentage of fibers containing type I myosin heavy chains and an increase in the percentage of muscle fibers containing type IIa myosin heavy chains. PMID- 17131835 TI - [Content of myosin-activating protein kinases in myocardium of patients with dilated cardiomyopathy and in the animal heart]. AB - The skeletal myosin light chain kinase (skMLCK) was identified in human and chicken embryo myocardium but not in embryo and adult rat heart using western blotting. The content of skMLCK and myosin-activating protein kinases: RhaA activated protein kinase (ROCK), integrin-linked protein kinase (ILK), and zipper interacting protein kinase (ZIPK) was compared in normal human myocardium and the hearts of patients with dilated cardiomyopathy (DCM). It was demonstrated that the content of skMLCK, ROCK and ILK increases in DCM whereas the content of ZIPK decreases. The results obtained may reflect compensatory processes in cardiomyocytes in DMC, which are aimed at increasing their viability and contractility. PMID- 17131836 TI - [Seasonal changes in phosphorylation of myosin regulatory light chains and C protein in myocardium of hibernating ground squirrel Citellus undulatus]. AB - A comparative study concerning the extent of phosphorylation of myosin regulatory light chains and C-protein from the left ventricle of hibernating ground squirrel Citellus undulatus during the periods of hibernation and activity was carried out. During hibernation, regulatory light chains of ground squirrel were found to be completely dephosphorylated. In active animals, the share of phosphorylated light chains averages 40-45% of their total amount. The extent of phosphorylation of the cardiac C-protein during hibernation is about two times higher than that in the active state. Seasonal differences in phosphorylation of the two proteins of ground squirrel myocardium are discussed in the context of adaptation to hibernation. PMID- 17131837 TI - [Plasticity of allogenic muscle tissue transplanted into a traumatized rat muscle subjected He-Ne laser radiation]. AB - The effect of muscle tissue alloplasty and laser radiation on the post-traumatic regeneration of skeletal muscles was studied. The implantation of an unirradiated minced allogenic muscle tissue into the area of trauma of a previously laser irradiated muscle had a positive effect. The allogenic muscle tissue underwent active regeneration. The traumatized muscle fibers in the rat-recipient muscle grew into the alloplant. The implantation of a previously laser-irradiated muscle tissue into the area of trauma of an unirradiated muscle was less effective. The allogenic muscle tissue prevented the growth of rat-recipient myofibers and underwent lysis in general. The allotransplantation of a laser-irradiated muscle tissue into an x-irradiated (20 Gy) traumatized muscle increased the destruction of rat-recipient muscle tissue. PMID- 17131838 TI - [The absence of support loading on the rat m. soleus leads to an increase in the number of intrafusal muscle fibres in muscle spindles]. AB - The ultrastructure of muscle spindles (incapsulated mechanoreceptors of stretch of extrafusal muscle fibres) of m. soleus in adult Wistar rats after repeated unloading of support on hind limbs with preservation of support loading on fore limbs has been studied by transmissing electron microscopy. It was shown that, along with muscle spindles with the ordinary number of intrafusal muscle fibres (four), m. soleus contains spindles with an increased number of intrafusal fibers (five to six). It was assumed that the increase in the number of intrafusal muscle fibers is due to the proliferation of their satellite cells. PMID- 17131839 TI - [Biological activity of electrochemically activated solutions obtained in a diaphragm electrolyser]. AB - The biological activity of the catholyte and anolyte of bidistilled water in experiments with the germination of wheat grains in the period from March to May has been studied. The activity of solutions, which was characterized by the grain germination index, was high at the beginning of the period, then it gradually decreased and was equal to zero at the middle of the period; at the end of the period it gradually increased almost to initial values. It has been established that the effectiveness of bidistilled water anolyte was as a rule higher than that of catholyte throughout the observation period. At the beginning and end, the stimulating effect of anolyte was 5-5.5 times greater than that of catholyte. The seasonal changes in the biological activity of M 9 medium catholyte were compared with those of bidistilled water anolyte and catholyte. The stimulating effect of M 9 catholyte was estimated by changes in the growth of E. coli cells. The stimulating effect, which was estimated from an increase in the optical density of cell suspension in the initial period at a cultivation temperature of 20 degrees C was 55-60% relative to control (untreated medium). Then it decreased almost to zero in the middle of the period to increase again approximately to the initial values. The assumption has been made that the physicochemical causes of the influence of catholyte and anolyte of bidistilled water on wheat grains and of the culture medium catholyte on E. coli cells are of different nature. PMID- 17131840 TI - [Titin isoforms]. AB - The titin isoform composition in skeletal and cardiac muscles of humans and animals has been studied using SDS elecrtophoresis in agarose-strengthened 1.3 2.3% polyacrylamide gels modified by us and immunoblot analysis in order to reveal new titin isoforms with molecular weights of more than 3700 kDa. The experimental data obtained have provided a basis for the suggestion that the titin bands of high molecular weights discovered by us are intact titin isoforms, while the titin bands designated on electrophoregrams as N2A, N2B and N2BA isoforms are their fragments. PMID- 17131841 TI - [International Symposium "Biological Motility: Basic and Application Research", Pushchino, Moscow Region, May 11-15, 2006]. PMID- 17131842 TI - [Atherogenic trans isomers of fatty acids in some food products]. AB - OBJECTIVE: Trans isomers of fatty acids (TFA) contain double bonds in their molecular structure with the trans configuration. Food products prominent for their TFA content include plant oils (margarine), mayonnaise, products baked with plant oils, and ready-made glazes and icings. The so-called fast foods and snacks (bars, chips) represent a significant source of TFA, particularly for children and youth. Considering the potentially atherogenic properties of TFA, this work was undertaken to determine the content of the principal monounsaturated TFA in some food products ubiquitous on the market, i.e. margarines and snacks. MATERIAL AND METHODS: We studied bars (Snickers, Mars, Twix), chips (Lay's, Snack, Cheetos), and margarines (Rama Classic, Rama Olivio, Vita, Flora, Finea, Delma, Masmix, Krowka, Smakowita with butter, Extra Pomorskie). The content of TFA was determined with gas chromatography. RESULTS: The highest content ofmonounsatured TFA among sweets was disclosed for the Twix bar (2.94 microg/g), while the lowest for Snickers (0.3 microg/g). Among margarines, the highest content of TFA was found for Smakowita with butter (87.6 microg/g), Krowka (46 microg/g), and Vita (38.7 microg/g), whereas the content was lowest in Finea (0.12 microg/g) and Delma (0.14 microg/g). In the case of chips, the content was highest for Cheetos (3.5 microg/g) and lowest for Snack (0.09 microg/g). CONCLUSIONS: The content of atherogenic TFA in common food products varies widely. In some cases, the content of TFA is considerable and unjustly undisclosed by manufacturers. Promotion of these products should include information in respect of TFA. This conclusion is of particular importance considering the significance of these products in the diet of children and youth. PMID- 17131843 TI - [The influence of the Metarhizium fungi on the embryonic development of Ascaris suum]. AB - PURPOSE: The present study focused on the effect of two fungal species Metarhizium flavoviride and Metarhizium anisopliae on the embryonic development and viability of Ascaris suum larvae. RESULTS: Microscope examination revealed that the presence of the mycelium inhibited the egg development, compared to control. The fungus-exposed eggs featured zygote vacuolisation, irregular blastomere cleavage, and morphological disturbances in the stages of blastula, gastrula, and larva. The embryopathies were significantly more frequent in the eggs incubated with M. flavoviride. Also the mortality of the developed larvae was significantly higher after 60-day culture with M. flavoviride. PMID- 17131844 TI - [Optimized diagnostic performance of brain magnetic resonance imaging in children with idiopathic growth hormone deficiency]. AB - PURPOSE: The aim of this study was to search for correlations between anatomic changes in the pituitary gland and hormonal disturbances in children with short stature. MATERIAL AND METHODS: Children with short stature were enrolled when criteria of pituitary growth hormone deficiency were partly or completely met. Magnetic resonance imaging was performed in 87 children and particular attention was given to the pituitary gland. Measurements were compared with pituitary dimensions accepted as normal in the literature. Contrast with GdDTPA was used to visualize the pituitary gland and associated structures (stalk, infundibulum). T1 weighted images in the sagittal and coronal planes were obtained. The results were statistically analyzed with non-parametric tests. CONCLUSIONS: 1. Magnetic resonance imaging is a very sensitive method for detecting changes in the pituitary gland and may well be recommended as a method of choice even though the percentage of changes detected with it is rather small. 2. The use of contrast agent may be abandoned to limit costs when searching for cause of growth deficit in children with idiopathic growth hormone deficiency, save for the following cases: hypoplasia or aplasia of the pituitary gland, transection of the stalk, empty sella syndrome or tumor in the central nervous system. 3. Pituitary volume and height appear to be of greatest diagnostic significance, while width (which varies little) can serve as an auxiliary parameter. PMID- 17131845 TI - [Contemporary applications of infrared imaging in medical diagnostics]. AB - INTRODUCTION: Thermal imaging is a non-contact, non-invasive diagnostic method for study human body temperature. Therefore infra red thermal imaging finds increasing application in clinical medicine. PURPOSE: The aim of this paper was to present and discuss the history and applications of thermal imaging in medicine. MATERIAL AND METHODS: The literature dealing with the history and applications of thermal imaging in medicine has been reviewed. RESULTS: Medical thermography was born in 1957 when a surgeon, Dr. R. Lawson discovered that his breast cancer patients had higher skin temperature over the cancer area. Since the 1970's thermography has been used in many areas of medicine. Early problems such as low detector sensitivity, but most significantly, poor training ofthermography technicians was the source of error in thermography and retarded the acceptance of this technique until 1990. Since that time, thermographic equipment has evolved significantly. Modern thermal imaging systems comprise technically advanced thermal cameras coupled to computers with sophisticated software solutions. The recorded images are now of good quality and may be further processed to obtain reliable information. Thermography can be applied as a diagnostic tool in oncology, allergic diseases, angiology, plastic surgery, rheumatology, and elsewhere. Contemporary thermal imaging must be per formed according to certain principles aimed at reliability and reproducibility of results. CONCLUSIONS: 1. Thermography is a safe, accurate and, most importantly, a non-invasive diagnostic method in clinical medicine. 2. Ignoring any of the principles worked out by the European Association of Thermology leaves thermography open to error and thus reduces acceptance of this technique in medical diagnostics. PMID- 17131846 TI - [Aplication of magnetostimulation in JPS system of chronic periapical tooth inflamation visible in the RTG picture]. AB - PURPOSE: The aim of the work was to use the magneto-stimulation in JPS system as a therapy aiding the treatment of chronic inflammation of periapical tissues of a tooth and to evaluate its effectiveness in RTG in Digora 2.1 digital system and in conventional RTG. MATERIAL AND METHODS: The research was based on 62 teeth of 56 people (some treated for the first time). There was also a comparative group. In the work the magnetic field was used since the first day ofendodental treatment. Every patient had RTG done before and after the magnetostimula-tion. The qualitative and quantitative evaluations were done. The optical density both in the area of the osteolisis and in the laudable bone was presented in tables and graphs. The statistical analysis was also done. Conventional RTG was also used to evaluate the research. CONCLUSION: It was ascertained that magnetic field of low induction has a positive effect on the regeneration processes in chronic inflammations of periapical tissues of the tooth, visible in RTG. PMID- 17131847 TI - [The hemostasis parameters and ACE gene polymorphism in patients with essential hypertension treated with perindopril]. AB - PURPOSE: 1. The assessment in haemostasis system, fibrinogen, tissue plasminogen activator (t-PA) and its inhibitor type-1, von Willebrand factor and beta thromboglobulin (beta-TG) levels and the changes in renin-angiotensin system (RAS), by determined angiotensin converting enzyme I (ACE) activity, in patients with essential hypertension (in relation to left ventricular hypertrophy) in comparison with normotension subjects. 2. The analysis of changes in haemostasis and RAS during treatment with perindopril in relation to left ventricular hypertrophy (group LVH+ and LVH-). 3. The assessment of haemostasis parameters levels and ACE activity in effect of treatment with perindopril in relation to ACE gene I/D polymorphism. MATERIAL AND METHODS: The study involved 78 male divided on two groups: cases and controls. The cases contained 44 male outpatients (25-38 years old) with untreated essential hypertension without clinical feature of coronary heart disease and 34 age- and gender- matched healthy controls. CONCLUSION: 1. Untreated essential hypertension predisposes to the procoagulant state characterised by increased Fb level and suppressed of fibrinolysis. These changes not depend on ACE gene I/D polymorphism. The left ventricular hypertrophy not intensificates the procoagulant state. 2. The treatment with perindopril decreases blood pressure effectively independent of ACE gene I/D genotype and left ventricular hypertrophy. Perindopril impairs RAS activity, most in patients with ACE gene II genotype. The treatment with perindopril is only partially effective in alleviating the procoagulant state, by reducing fibrinogen level in II homozygotes due to its more potent inhibitory action on the RAS in this group. The treatment makes fibrinolysis better mainly to high t-PA levels, independently of ACE gene I/D polymorphism. 3. Left ventricular hypertrophy not differentiates the response on treatment in relation to haemostasis parameters and RAS, however there was a slight decrease in ACE activity in patients with left ventricular hypertrophy. 4. The changes in haemostasis system and ACE activity, observed during treatment, suggest the appearance of positive relation between haemostasis and RAS. Probably the antithrombotic act of perindopril is more pronounced in II genotype group. PMID- 17131848 TI - The influence of low birth weight on the prevalence of refractive errors among schoolchildren. AB - PURPOSE: The aim of the study was to determine in a large population whether low birth weight has an influence on the prevalence of refractive errors among schoolchildren. MATERIAL AND METHODS: 3663 schoolchildren were examined (1738 boys and 1925 girls, aged 6-17 years, mean age 11.1, SD = 3.2). The weight at birth was 1500-2500 grams (mean 2184, SD = 271) in 254 and more than 2500 grams (mean 3398, SD = 441) in the remaining 3409 children. Skiascopy with cycloplegia was done and refractive error readings were reported as the spherical equivalent (SE). Myopia was defined as SE < or = -0.5 D, hyperopia as SE > or = +1.5 D. Anisometropia was diagnosed when the difference in the refraction of both eyes was > 1.0 D. The parents completed a questionnaire on the child's weight and term of birth. Data analysis was performed using chi-square test. P values of less than 0.05 were considered statistically significant. RESULTS: Hyperopia was observed more frequently in 6-7 year-old children whose birth weight was > 2500 grams (p < 0.05) - table 1. Additionally, it was found that anisometropia was less frequent in 10-11 year-old children whose birth weight was > 2500 g (p < 0.05) - table 2. CONCLUSION: Low birth weight may have an effect on the prevalence of refractive errors among schoolchidren. PMID- 17131849 TI - [Retinopathy of prematurity--pathogenesis and prevalence]. AB - INTRODUCTION: Retinopathy of prematurity (ROP) is a retinal disease in premature newborns. The etiology of this disease is complex and not fully understood. For many years, ROP was the main cause of blindness and impairment of visual acuity in prematurely born children in Poland. Progress in neonatal medicine helps save the life of increasingly more neonates with extremely low birthweight but at the same time ROP is observed more often. Today, ROP has grown to the chief problem in contemporary pediatric ophthalmology. Therefore, it is crucial from the medical and economic point of view to understand, prevent, and manage ROP. CONCLUSIONS: The present study focuses on the epidemiology, etiology, and pathogenesis of ROP. Information on ophthalmologic screening in Poland is also provided. PMID- 17131850 TI - [Retinopathy of prematurity: clinical findings and current opinions on diagnosis and treatment]. AB - INTRODUCTION: Retinopathy of prematurity (ROP) is an ophthalmologic condition which may lead to blindness if compounded by other risk factors. Since many years, ROP is among the main causes of impaired visual acuity and blindness in Polish children. Therefore, the understanding of this disease, progress in diagnostics, and improved methods of treatment will contribute to a better prognosis in ROP. Nowadays, diode laser photocoagulation and cryotherapy are modalities used in ophthalmology centers at specific clinical stages of ROP. When performed at the right time, these procedures offer promising results in the re attachment of the retina and restoration of vision. Interventions for the most advanced vaso-retinal changes are rare because of a poor outcome. Early detection of vascular and retinal changes during ROP is burdened by some technical difficulties. Therefore, classification of findings in ROP should be done by ophthalmologists specialized in this area. The role of the neonatologist is also important, particularly when it comes to awareness of the parents as to the treatment needed by their prematurely born child. Laser photocoagulation, cryotherapy, and surgery undertaken in ROP may lead to ophthalmologic complications, with refraction errors and eyeball mobility disorders being the most common. Consequently, it is important for the child to undergo regular ophthalmologic examinations. PMID- 17131851 TI - [The impact of education in birthing schools on the course of pregnancy, labor, puerperium, and neonatal care]. AB - INTRODUCTION: Pregnancy represents a special period in the life of a woman. It is the time of many joyous, sublime, and touching moments experienced by the family although not free from anxiety, uncertainty and doubt in the minds of the future parents. It is also the time of an intense quest for answers to annoying questions and a motivating factor to search for information on the physiology of pregnancy, as well as on labor, puerperium and child care. A birthing school is nowadays the most important institution which prepares for parenting in a most professional way. This form of education is gaining widespread acceptance. This study was designed to: 1. Study the effects of participation in birthing school classes on the course of pregnancy, labor, and puerperium. 2. Search for relationships between the birthing school program and motivation of mothers to breast feeding. 3. Determine the influence of education in a birthing school on the ability of mothers to cope with problems during puerperium. MATERIAL AND METHODS: The study was done in 294 women hospitalized at the Obstetrics Ward of the Chair and Department of Obstetrics and Perinatology, Pomeranian Medical University in Szczecin, between June 2001 and December 2002. Two groups were formed: group I--"study group"--comprised 129 gravida who participated in birthing school classes during at least one pregnancy but not earlier than two years before the present study; group II--"control group"--included 165 gravida who never participated in any organized form of birthing education. A questionnaire was developed consisting of 105 questions. Answers were recorded by the researcher and the results in both groups were compared statistically. CONCLUSIONS: Participation in birthing school classes helps develop healthy attitudes during pregnancy, reduced the feeling of pain and blood loss during labor, but does not alter the duration of labor in primipara nor has any effect on problems related to puerperium. Graduates of birthing schools are better motivated to start breast feeding, less often switch to artificial feeding during puerperium, and cope better with problems encountered during puerperium. PMID- 17131852 TI - [Medical-social aspects pregnancy, labor and puerperium in patients the solitary mother home in Karwowo near Szczecin]. AB - PURPOSE: Solitary maternity is a significant problem of modern times requiring socio-economic and psychological, as well as perinatological studies. The twentieth anniversary of the Maternity Home managed by the Benedictine Samaritan Sisters motivated the present study. MATERIAL AND METHODS: Hospital discharge summaries of 429 women (group DSM) who lived at the Maternity Home between 26 July 1983 and 26 February 2002 were analyzed. This documentation was supplemented with hospital data on 177 women who gave birth at two Szczecin hospitals and on 13 who left the Home before delivery. RESULTS: Residents of the Home came from all regions of Poland and from urban and rural societies. They were between 14 and 43 years of age (mean 22.6 +/- 5.7). 87.4% of them were single and 63.7% were primipara. 50.4% of them failed to complete primary education while 3.4% had university-level education. 6% were mentally and/or psychologically handicapped and 42.7% had no profession. The major causes of admission into the Home were: desire to hide the pregnancy, family conflicts, homelessness, difficult material situation, mental or physical disability, violence or alcoholism in the family. The control group comprised 400 women (group K) who gave birth at the Department of Obstetrics and Perinatology, Pomeranian Medical University in Szczecin. 51.5% of them were primipara. The mean age in this group was 27.4 +/- 5.4 (17-44). 71.3% were married. The Majority had secondary (41.9%) or university (32.5%) education. Most of them worked as artisans, teachers or economists. Neurological or psychiatric diseases dominated in group DSM but otherwise no severe disease at all could be disclosed. Satisfactory prenatal care was provided to 33.1% of DSM patients and to 78% of group K. No prenatal care was provided to 9.5% of DSM patients and to 1.7% of group K. EPH gestosis was more often observed in DSM patients and imminent abortion, premature birth, PROM, urinary tract infection, and anaemia were more frequent in controls. More DSM patients had no treatment during pregnancy than controls. The use of tocolytics was significantly more frequent in DSM patients. Antibiotics and drugs accelerating maturation of the respiratory system were applied with similar frequency in both groups. Frequency of immature and mature births was similar in both groups. 84.2% of DSM patients and 68.8% of controls had natural delivery. Cesarian section was twice more frequent in group K. Immediate indications dominated in both groups (74.1% in DSM; 57.4% in K). No medication was used during delivery in 30% of patients. DSM patients more often required strong analgesics. Antibiotics and agents increasing uterine tension more often were used in K patients. Deliveries in both groups were usually without complications. Female genital tract injury was more frequent in DSM patients (20.9% in DSM; 14.8% in K). Delivery duration was similar in both groups, but the second part of delivery was longer in K primipara. Mean neonatal weight in the 1000-2500 g range was similar in both groups but was significantly higher in controls for 2501-4000 g neonates. The difference for newborns weighing more than 4000 g was not significant. There were no neonates weighing less than 1000 g. The clinical condition of newborns measured with the Apgar scale was worse in the DSM group after the 1st min, as well as after the 3rd and 5th min and the difference in each case was significant. Puerperium was usually uncomplicated. However, urinary tract infection was more frequent in DSM patients and anaemia was more frequent in controls. The duration of hospitalisation was similar in both groups (mean = 6 days). 120 of 177 DSM mothers left hospital together with their children, as compared to 99.3% in the control group. 87 children of mothers from the Maternity Home were offered for adoption. CONCLUSION: By studying unwed maternity it was observed that a holistic approach is the only way to disclose all the problems which are often unusually complex and concealed but which determine the normal development of an individual in the society. Analysis of the factors involved may help in attaining goals that serve the good of the society. PMID- 17131853 TI - [The dilatation of auditive tube (Bougirage tubaire) by dilators in XIX century]. AB - The treatment of auditiory tube chronic catarrh in XIX century by pharmagological drugs, by special apparatuses of surprising technical sophistication, by electrisation and faradisation is discussed first of all. The dilatation of auditory tube by dilators was the diagnostical and therapeutical methods, especially preferenced by Victor Urbantschitsch, Hermann Schwartze, Jean P. Bonnafonte, Albert Calmettes and Bronislaw Taczanowski, Teodor Heiman and Samuel Meyerson. The dilators were building of celluloid, whale-bone, silver and were absorbed by argentum nitrate, vaseline. The technique, difficulties and effects of dilatation are described in more detail. The complications of this operation is presented finally. PMID- 17131854 TI - [Influence of psychological therapy on duration of survival in patients with metastatic cancer]. AB - The work of A. J. Cunningham has created new perspectives for extension of survival through psychological intervention in patients with advanced cancer. Therapeutic techniques exploiting unique subjective psychic and social experiences may be particularly helpful at the time when life of the patient is threatened by an incurable disease. Psychological therapy should be based on an individual plan for living with cancer. Novel qualitative research and therapeutic methods need to be developed. PMID- 17131855 TI - [Explosion of the subjective world]. AB - PURPOSE: This text was inspired by an article in psycho-oncology: "The influence of psychotherapy on the extension of life in people with advanced cancer". I intend to show that cancer causes disintegration ("explosion") of a person's subjective world. MATERIAL AND METHODS: (philosophical discourse): Categories of the personal world and the subjective world are introduced and the reality of the subjective world is accepted. The following connotations are essential to further discussion: The mind acts on the body. The psychic state (therefore the state of the brain) is reflected in states and processes localized outside the brain. The epistemologist desires to discover the causal mechanism that explains these effects. The body may act on the mind. Overwhelming evidence has accumulated that disorders of the mind may be caused by disturbances of processes in the nervous system. I reject the view that "the brain is the cause of the mind". CONCLUSIONS: Every person lives in his or her own, private, subjective world. It is a relatively stable world of personal memories, observations and imaginations. This world extends in subjective time and space. Its content is composed of conscious psychic experiences, one after another. Experiences create the stream of consciousness. Serious illness leads to the explosion or disintegration of subjective world, destruction of one's sense of personal identity, impossibility of autoreflection, and loss of sense of living. In such cases, individualized psychotherapeutic intervention proves to be irreplaceable. PMID- 17131856 TI - [Morphology of the glottis after supracricoid laryngectomy with CHP or CHEP and restoration of basic laryngeal function with special attention to quality of phonation]. AB - PURPOSE: The aim of this work was to describe the morphology of the reconstructed larynx after supracricoid horizontal laryngectomy with simultaneous cricohyopexy (CHP) or cricohyoepiglottopexy (CEHP) preserving one or both arytenoid cartilages, to study the protective and respiratory functions of the larynx, to perform phoniatric evaluation of phonation quality, to analyze voice and speech phonetic-acoustic parameters, and to search for correlations between reconstruction type and phonetic-acoustic characteristics of voice and speech. MATERIAL AND METHODS: The material comprised 58 patients (32 after CHP and 26 after CHEP). CONCLUSION: Oval shape of the neoglottis dominated. Swallowing was normal in 46% of patients and did not depend on reconstruction type. Decannulation was possible in 66% of patients. The results were inferior after CHEP with both arytenoid cartilages intact. Socially efficient speech was found in 74% of patients and the results were better with CHEP. The phonetic-acoustic structure of voice and resonant speech was considerably different from the phonetic-acoustic structure of voice and speech under physiologic conditions. These differences applied to segmental (formant structure frequencies, noise range), as well as suprasegmental voice features. PMID- 17131857 TI - Intracerebral haematoma in hydrocephalic shunted patients not related to ventricular cannula. Report of three cases and review of the literature. AB - Intracerebral haematoma (ICH) distantly from the site of insertion of a ventriculoperitoneal shunt or external drain is an extremely rare complication. In contrast to ICH caused by disruption of small cerebral vessels by the catheter, the cause of haematoma distantly from the shunt is unclear. We report three cases of ICH occurring distantly from the tubing path. We also discuss possible mechanisms and provide a literature review of this complication. PMID- 17131858 TI - [Indication and limitation of office surgery: Office surgery of the ear- indication and limitation]. PMID- 17131859 TI - [Clinical investigation using a self-rating depression scale in subjects with psychosomatic taste disorders]. AB - In recent years, the number of patients with psychologenesis taste disorders has increased. In this study, related psychological factors were examined in 258 patients (85 men, 173 women) using a self-rating depression scale (SDS). The results were as follows: 1) 189 patients reported normal psychological factors, 2) 173 patients reported neurotic sates, and 3) 40 patients reported positive correlations with a depressive sates, Most of the patients, self-reported results were not correlated with taste function tests. On the other hand, the patients with depression had reduced salivary secretion tests results. Only 50% of the patients with depression recovered their taste functions. These results suggest that the SDS test may be useful for diagnosing taste disorders and monitoring effects of treatment. PMID- 17131860 TI - [Epidemiological survey of allergic diseases in first-year junior high school students in Wakayama Prefecture in 2003]. AB - Allergic diseases have been increasing in prevalence in developed countries, including Japan. The aim of the present report was to determine the prevalence of allergies in Wakayama prefecture using an epidemiological study. In total, 759 first-year students attending junior high school in Hidaka country, Wakayama prefecture, were surveyed. The results for 699 cases were then analyzed. A questionnaire regarding allergic diseases, specific IgE measurements performed using a MAST26 system (Hitachi Co., Ltd.), and total serum IgE levels measured using RIST (Pharmacia Co., Ltd.) were performed. The prevalence (present + past) of various allergic diseases was 37.9%. The prevalence of rhinitis, including pollinosis, was 31.0%, while that of atopic dermatitis was 26.2% and bronchial asthma was 11.3%. The positive rates for specific IgE antibodies against Japanese cedar pollen was 48.6%. The positive rate for Dermatophagoides farinae, timothy and housedust II were 44.2%, 29.6%, and 28.9%, respectively. Statistically significant differences were recognized between the students with allergic rhinitis, atopic dermatitis, allergic conjunctivitis or bronchial asthma and positive results for D. farinae, housedust II, cedar pollen, Penicillium, Cladosporium, or Aspergillus-specific antibodies. Regarding family history (two generations), a statistical significant difference between family history and positivity for specific IgE antibodies like D. farinae, housedust II, ragweed, cedar pollen or Cladosporium was observed. The total IgE titer was correlated with the number of positive allergen items. The increasing prevalence of various allergic diseases in developed countries remains a mystery, but the hygienehypothesis has attracted some attention. The findings of this epidemiologic study will contribute to basic data on the increasing prevalence of various allergic diseases. PMID- 17131861 TI - [Open adrenalectomy for adrenal metastases from lung cancer--usefulness and problem of laparoscopic adrenalectomy]. AB - We examined the outcome of open adrenalectomy performed at our hospital to determine the effectiveness and problems of laparoscopic adrenalectomy for adrenal metastases from lung cancer. Between January 2001 and June 2004, eight open adrenalectomies were performed on six patients with adrenal metastases from lung cancer. Surrounding adhesion was observed in five of the eight cases (63%). The recurrence period was 17.8 months and 5.3 months in the cases with and without adhesion, respectively. Laparoscopic adrenalectomy for adrenal metastases may be safe and effective. However, potential surrounding adhesions meed to be adequately considered, if the primary lesion is the lung. PMID- 17131862 TI - [Malignant fibrous histiocytoma of the retroperitoneum--review of clinical course and histopathology]. AB - The definition of malignant fibrous histiocytoma (MFH) was modified and simplified in the new WHO-Soft Tissue and Bone Tumor Classification published in 2002. We reviewed the clinical courses of 7 patients with MFH of the retroperitoneum treated in our hospital from 1985 to 2005 and reexamined their pathological diagnoses according to the new classification. All pathological specimens were reviewed again by an expert pathologist (TH) and were confirmed as MFH. Median follow-up was 5 months (0.5 to 44 months). Five patients were diagnosed as having the pleomorphic type, and 2, the inflammatory type. The tumor was located in the retroperitoneal space in 6 patients and around the left spermatic cord in 1. Radical surgery was the primary treatment for 6 patients. The disease recurred locally in 5 patients and distant metastasis developed in 3. The lung and liver were the principal locations of distant metastases. The 2-year disease-specific survival rate was 30% and 2 patients have been alive for more than 3 years. Our experience and those of others reported in the literature suggested that the factors for long-term survival in MFH are pleomorphic-type histology, superficial location, small tumor size and intensive wide excision, although most patients with MFH might have a poor prognosis. Because the tumor size of MFH tended to be large, especially that in the retroperitoneal cavity, wide excision with sufficient tumor-free margin is hard to guarantee. PMID- 17131863 TI - [Hydrodistention of the bladder in patients with interstitial cystitis--clinical efficacy and its association with immunohistochemical findings for bladder tissues]. AB - We retrospectively analyzed the clinical relevance of hydrodistention under anesthesia for patients having urgency and/or lower abdominal pain who were clinically diagnosed as having interstitial cystitis (IC) from May 1996 to May 2005. Their symptoms were refractory to anticholinergic or antiinflammatory agents. Hydrodistention was performed under general or spinal anesthesia with direct vision by cystoscopy and irrigation fluid was instilled into the bladder at a pressure of 80 cmH2O. Cystoscopic findings revealed glomerulation in 26 patients (96%), cracking in 10 (37%) and Hunner's ulcer in 3. Twenty-four patients (89%) obtained improvement of the objective symptoms after treatment. However, symptoms soon deteriorated in 16 patients, and the average duration of efficacy was only 4.7 months (SD; +/-3.7). There were two episodes of complication in this treatment. Bladder rupture occurred during hydrodistention, but was successfully managed with simple percutaneous perivesical drainage. One patient with acute pyelonephritis was treated with an antimicrobial agent without any additional treatment. Although bladder specimens were examined by immunohistochemistry, tryptase and c-kit were not linked with the mast cell count, severity of symptoms or treatment efficacy. Hydrodistention of the bladder may be recommended as the first treatment choice for patients with IC because it provides relatively high efficacy. However, the short duration of the efficacy requires a second-line treatment option for better management of patients with IC. PMID- 17131864 TI - [Clinical study of 18 vesicointestinal fistulas]. AB - We studied 18 cases of vesicointestinal fistula surgically treated between January 2001 and July 2005. The underlying'cause was an inflammatory disease in 12 cases, a carcinoma in 5 and injury (post-radiation therapy) in 1 case. The fistula was visualized by cystography in 2 cases and enterography in 4. Surgical procedures were cystectomy with enterectomy in 2 cases, partial cystectomy with enterectomy in 3, bladder wall overlay-suture with enterectomy in 6 and enterectomy alone in 4. In 3 cases, colostomy without enterectomy was performed for palliative surgery. In all cases the postoperative course was good and surgical treatment was effective. Surgical procedures varied in each case depending on the etiology and the patient's condition. PMID- 17131865 TI - [Significance of re-measurement of prostate specific antigen before prostate biopsy]. AB - To examine the natural change of prostate-specific antigen (PSA), we compared two PSA values, the first value with which we decided to perform prostate biopsy and the value obtained by remeasurement just before biopsy. To exclude cases with extremely high PSA, we examined 288 cases in which PSA was less than 50 ng/ml for comparison. Of the 288 cases, 93 were diagnosed with prostate cancer (CaP). The interval between the two PSA measurements was 1-90 days (average of 31.4 days). The first and second values were an average of 13.0 and 11.7 ng/ml, respectively, and the second value was significantly lower than the first value. When we divided them into CaP cases and non-CaPthe two cases, a significant difference between PSA values was found only in the non-CaP cases. Moreover, in the non-CaP cases with some clinical symptoms, the difference in PSA was marked between the first and second values, which averaged 11.2 and 9.2 ng/ml, respectively. When we decide to perform a biopsy, we should recognize that PSA sometimes is lower on re measurement. Particularly in symptomatic cases, it is worth re-measuring PSA, which may save unnecessary biopsies. PMID- 17131866 TI - [Initial results of holmium laser enucleation of prostate for benign prostate hypertrophy patients]. AB - We have performed holmium laser enucleation of prostate (HoLEP) in 14 cases since February 2005. The patient's age ranged from 62 to 84 years old (average: 75). The preoperative results of International Prostate Sympton Score (IPSS), quality of life index, maximum urinary flow rate and residual urine volume were compared with those 3 months after surgery. The mean operation time, the mean estimated prostate volume and the mean enucleated prostate volume were 143 min, 50 cm3 and 35 g, retrospectively. All cases showed improvement in each parameter, and all but one case recovered continence in 3 months after surgery. A periurethral abscess had developed in the patient, suffering from incontinence. We concluded that HoLEP is an effective treatment. The relatively higher incidence of urinary incontinence in HoLEP remains to be resolved in the future. PMID- 17131867 TI - [Histological investigation of prostate cancer treated with hormonal agents]. AB - The histological diagnosis of prostate cancer treated with hormonal agents is often difficult because of various morphological changes induced by androgen ablation. Immunostaining of cytokeratins may be useful to prevent the underdetection of cancer cells. We examined prostatic specimens with histological diagnosis of 11 pTO patients who had undergone neoadjuvant endocrine therapy followed by radical prostatectomy. Anti-cytokeratin antibody, AE1/AE3 was used to detect the prostatic epithelial cells. Anti-cytokeratin antibody, 34/3 E12 was used to detect the prostatic basal cells. The loss of basal cells indicates the acini to be cancer. The immunostaining with these antibodies revealed that 2 out of 11 cases had residual cancer and were not pTO. The immunostaining of cytokeratins was useful to detect the residual prostatic cancer after endocrine therapy. PMID- 17131868 TI - [Hemorrhagic adrenocortical adenoma with myelolipoma: a case report]. AB - We present a case of hemorrhagic adrenocortical adenoma with myelolipoma. A 66 year-old woman was admitted to our hospital for left retroperitoneal mass. Based on abdominal computed tomography, magnetic resonance imaging and blood tests, preoperative diagnosis was a sarcoma of renal capsule origin. En bloc resection of adrenal gland, tumor, and the kidney with lymph node dissection was performed. Histologically, the mass was diagnosed as hemorrhagic adrenocortical adenoma with myelolipomatous foci. PMID- 17131869 TI - Immune thrombocytopenia during interferon-alfa therapy for renal cell carcinoma. AB - Immune thrombocytopenia is a rare complication of interferon-alfa (IFN-alpha). A patient with renal cell carcinoma developed severe thrombocytopenia during therapy with purified IFN-alpha. The patient's exposure to IFN, exclusion of other causes, and bone marrow biopsy were consistent with drug-induced immune thrombocytopenia. Cessation of IFN and corticosteroid administration resulted in the prompt recovery of platelets. The patient was re-challenged with recombinant IFN-alpha-2b under careful observation; there was no occurrence of severe thrombocytopenia. It was suggested that the difference of the subtypes composing IFN-alpha resulted in the lack of cross reactivity. PMID- 17131870 TI - [Ileovesical fistula caused by bladder cancer with bladder stone]. AB - Enterovesical fistula is a very rare complication of primary urological malignancies. A case of ileovesical fistula caused by a bladder carcinoma is presented. A 66-year-old male was referred with complaints of urinary pain. On admission, fecaluria and urinary tract infection with bladder stone were detected. Cystography revealed the passage of contrast medium into the small bowel. Under the diagnosis ofileovesical fistula due to suspected inflammatory disease, sigmoidectomy and segmental small bowel resection with partial cystectomy were performed. Histological evaluation revealed a poorly differentiated urothelial carcinoma. Without further treatment, the patient died from cancer five months after operation. However, it is hard to assess the effect of fistulas on prognosis. Since it has been reported that about 40% of the patients with T4 bladder tumors could be potentially cured with radical resection, we recommend a thorough examination to confirm the diagnosis of primary disease to obtain the best results. PMID- 17131871 TI - [A case of transitional cell carcinoma of the bladder in a juvenile patient]. AB - A 17-year-old male was referred to our hospital with the chief complaint of right back pain. Cystoscopic examination revealed a papillary tumor on the posterior wall following detection on screening ultrasound examination revealed a tumor in the bladder. Transurethral resection of the bladder tumor was performed. Histological examination of the excised tumor revealed transitional cell carcinoma, grade 1, pTa. No recurrence has been observed for about 1 year postoperatively. PMID- 17131872 TI - [Pelvic lymph node metastasis from bladder cancer markedly responsive to methotrexate vinblastine doxorubicin and cisplatin (M-VAC) therapy followed by radiotherapy: a case report]. AB - We report a case of bladder cancer with pelvic lymph node metastasis effectively treated by chemotherapy followed by radiotherapy. The patient was a 65-year-old man who had undergone radical cystectomy. Histological findings showed urothelial carcinoma, G3 > G2, pT1b. After 31 months, computerized tomography (CT) revealed a bulky tumor (7.0 x 5.6 cm) along the left pelvic wall, indicating pelvic lymph node metastasis. Five courses of chemotherapy consisting of M-VAC (methotrexate, vinblastine, doxorubicin and cisplatin) was performed. The size of the tumor was reduced to 1.5 x 1.0 cm. Then, external beam radiotherapy (50 Gy) was added to the residual tumor. He has been alive with no evidence of disease progression for 31 months since the radiotherapy. PMID- 17131873 TI - Bladder perforation of the tension-free vaginal tape detected with a flexible cystoscope. AB - A 67-year-old woman who presented with stress urinary incontinence underwent a tension-free vaginal tape (TVT) operation. Postoperatively, she complained of urgency, gross hematuria and dysuria, which persisted for one week. Cystoscopy with a flexible cytoscope revealed penetration of the bladder wall by the tape, which had been missed during the TVT procedure by rigid cystoscopy. After the penetrating portion of the tape was excised, her symptoms were resolved and continence was preserved. This case suggests that employment of a flexible cytoscope is useful to detect the placement of tape in the bladder wall, which may be missed with a rigid cytoscope. PMID- 17131874 TI - [Small cell carcinoma of the prostate: a report of three patients and a prognostic analysis of cases reported in Japan]. AB - Small cell carcinoma (SCC) originating from the prostate is rare. We report three cases of SCC of the prostate. Case 1: A 29-year-old man with large pelvic mass and pelvic lymph node metastases was diagnosed as having pure SCC of the prostate. Chemo-radiotherapy resulted in a great reduction of the tumor volume. However, the disease recurred immediately, and he died of disease 17 months after diagnosis. Case 2: A 65-year-old man presented with pure prostatic SCC with lung metastases. Although cystoprostatectomy combined with pre- and post-operative chemotherapy ended with no evidence of disease, he died after 16 months because of multiple metastases and local recurrence. Case 3: A 73-year-old man was diagnosed as having SCC and poorly differentiated adenocarcinoma of the prostate simultaneously. Chemo-endocrine therapy and pelvic irradiation were performed, achieving partial remission. However, he developed multiple distant metastases, and died of disease 15 months after diagnosis. We reviewed 82 cases previously reported in Japan. Patient's ages ranged from 24 to 86 years (mean 68.7 years). Many patients had lymph node or distant metastases (stage D, 73%). Thirty-seven (45%) were pure SCCs and 45 (55%) were associated with adenocarcinoma. The prognosis after the recognition of SCC is very poor, and the 1- and 2-year survival rates were 27% and 10%, respectively. Survival did not differ in patients with pure SCC or mixed glandular and small cell carcinoma. Higher elevation of pretreatment serum NSE value was associated with the poor prognosis. PMID- 17131875 TI - [A case of granulomatous orchitis]. AB - A 66-year-old man came to us with a chief complaint of painful right scrotal swelling. The right testis was enlarged on the ultrasound gray scale, but the inside was homogeneous. A power Doppler ultrasound showed slightly increased vascular flow in the periphery of the right testis. As the soluble IL-2 receptor was 527 U/ml (normal: 220-530 U/ml), we considered malignant lymphoma and performed high orchiectomy. On pathological examination, the tumor was diagnosed as granulomatous orchitis. To our knowledge, this is the 21st case in Japan. PMID- 17131876 TI - [Synaptic plasticity regulates alternative 3'-end processing that modulates the production of alternatively spliced RNA]. PMID- 17131877 TI - [Local translational control at synapse]. PMID- 17131878 TI - [Novel mechanisms in signal transduction and cellular response through neural adaptor proteins]. PMID- 17131879 TI - [Physiological roles of nuclear CaM kinase II delta in the central nervous system]. PMID- 17131880 TI - [Transcriptional regulation of synaptic proteins by presenilin/gamma-secretase]. PMID- 17131881 TI - [Neuronal cell death and stress response]. PMID- 17131882 TI - [Structure and function of the presynaptic active zones]. PMID- 17131883 TI - [RPEL family proteins: transcriptional co-activators and mediators for actin based structural alteration]. PMID- 17131884 TI - [Role of activity-dependent gene expression of BDNF gene in synaptic plasticity]. PMID- 17131885 TI - [Diagnostic imaging in the field of anesthesiology: preface and comments]. AB - In this special issue, the diagnostic imaging in the field of anesthesiology is featured. Four topics including preoperative upper airway radiograph, perioperative bronchofiberscopy, transesophageal echocardiography (TEE), and perioperative diagnosis of deep vein thrombosis are discussed. Diagnostic imaging can be a powerful tool to improve the quality of perioperative management of surgical patients. PMID- 17131886 TI - [Recent advances in TEE during cardiac surgery]. AB - Transesophageal echocardiography (TEE) has opened a new window to the heart not only in adult cardiac surgery but in pediatric cardiac surgery. Its success rate in clinical decision making is impressive. It has become an important part of pediatric and adult cardiology. The combination of unique morphological and hemodynamic information of exceptional diagnostic quality and the possibility to use this diagnostic method as a bedside tool in the intensive care unit and the operating room guarantee that this diagnostic modality will maintain its firm place in the diagnostic armamentarium of cardiologists and anesthesiologists. TEE can provide an immediate gauge of surgical results and can be of help to avoid suboptimal surgical outcomes. We showed a recent advance with TEE modality including 3D TEE. PMID- 17131887 TI - [Potential roles of preoperative upper airway radiographs in planning of perioperative airway management]. AB - Majority of severe anesthetic complications result from airway difficulties. Among the airway difficulties, difficult mask ventilation is a key critical condition in the currently proposed difficult airway algorithms, and is probably the most life-threatening event. Presence of severe obstructive sleep apnea may indicate potential difficulty in mask ventilation during anesthesia induction. Lateral neck radiographs for patients with suspected airway difficulties may provide useful information for identification of potential patients with difficult mask ventilation. Among the parameters measured by the radiographs, I recommend anesthesiologists to measure distance between the hyoid bone and mandibular plane, which possibly reflects anatomical balance of the upper airway maintenance. PMID- 17131888 TI - [Diagnosis of the laryngeal or tracheobronchial lesions by fiberoptic bronchoscopy for anesthesiologists]. AB - Flexible fiberoptic endoscopic equipment is a useful visualization aid for diagnosis of laryngeal or tracheobronchial lesions and for intubation in patients with difficult airway. It also determines double-lumen tube position. In an intensive care unit, fiberoptic bronchoscopy is the cornerstone of the causal diagnosis in acute respiratory failure and laryngo-tracheobronchial trauma. However, for many anesthesiologists, its use tends to be limited to fiberoptic intubation and anesthetic management of thoracic surgery. Therefore, this review focuses on diagnostic strategies of laryngeal or tracheobronchial lesions by fiberoptic bronchoscopy for anesthesiologists. It also refers to the equipment and the strategy in performing fiberoptic bronchoscopy for anesthesiologist. We anesthesiologists need to attain proficiency in diagnostic skill in fiberoptic bronchoscopy as specialists of airway managent in acute settings as well as the operating room. PMID- 17131889 TI - [Diagnosis of deep vein thrombosis in the perioperative period]. AB - Deep vein thrombosis and pulmonary thromboembolism have been common illness even in Japan and are receiving increased social as well as medical attention, especially during postoperative period. Vital Statistics of Japan compiled by Ministry of Health, Labor and Welfare indicated that the number of patients who died of pulmonary thromboembolism had increased about ten-fold in this half a century. The most effective way of reducing unexpected death from venous thromboembolism is to institute a comprehensive institutional policy of primary prophylaxis and early diagnosis in patients at risk. The diagnostic strategy has changed along with the improvement especially of venous ultrasound sonography and multidetector-raw CT. The diagnosis of venous thromboembolism is now more noninvasive and accurate. However, we still have no screening test, although symptoms and signs of venous thromboembolism are more complex in perioperative period. Therefore, we need to observe more carefully. We have to find venous thromboembolism among patients with single leg swelling and pain, or sudden dyspnea and transient loss of consciousness, and link to this finding diagnostic algorithm. PMID- 17131890 TI - [Plasma levobupivacaine concentrations following epidural administration of levobupivacaine conjugated with or without maltosyl-beta-cyclodextrin]. AB - BACKGROUND: To investigate the pharmacokinetics of complexation of levobupivacaine with maltosyl-beta-cyclodextrin, the plasma concentrations of levobupivacaine were measured following epidural administration of levobupivacaine conjugated with maltosyl-beta-cyclodextrin or levobupivacaine alone in a rabbit model. METHODS: Rabbits were randomly divided into two groups, levobupivacaine (1%) group (n = 6) and levobupivacaine (1%) conjugated with maltosyl-beta-cyclodextrin (100 mM) group (n = 6). One ml of each solution was randomly injected through an epidural catheter placed at L5-6. The plasma levobupivacaine concentrations were measured before and 5, 10, 15, 30, 60, 120, 240, 360, and 480 min after injection. RESULTS: The plasma levobupivacaine concentrations were significantly higher in the levobupivacaine conjugated with maltosyl-beta-cyclodextrin group than in levobupivacaine group at 5 min (1,465 +/ 311, 1,033 +/- 347 ng x ml(-1)), 10 min (1,068 +/- 237, 731 +/- 191), and 15 min (958 +/- 311, 605 +/- 118). There were no differences in area under the curve (1,551 +/- 387, 1,176 +/- 154 ng x hr x ml(-1)) and elimination half life (100 +/ 54, 78 +/- 37 min) between the two groups. CONCLUSIONS: The results of this study indicated that the absorption of levobupivacaine conjugated with maltosyl beta-cyclodextrin from the epidural space and the elimination from the blood were similar to plain levobupivacaine. PMID- 17131891 TI - [General anesthesia for thyroplasty employing voice test]. AB - BACKGROUND: We reported an anesthetic technique for thyroplasty employing voice test. Thyroplasty is performed to improve voice quality in patients with unilateral vocal cord paralysis. Correct displacement of the vocal cord is assessed by asking the patient to phonate. At this point all patients should have recovered from general anesthesia and cooperate to phonation, facilitating correct displacement of the vocal cords. METHODS: Anesthesia was induced with i.v. pentazocine 15-30 mg and continuous propofol infusion 10 mg x kg(-1) x hr( 1). The patients received propofol infusion 4-6 mg x kg(-1) x hr(-1) with spontaneous ventilation. At the point of correct displacement of the vocal cords, we stopped propofol infusion and all the patients woke up immediately and cooperated. After determining the voice propofol was given at a rate of 4-6 mg x kg(-1) x hr(-1). RESULTS: The average times of surgery and anesthesia were 98 +/- 23 min and 139 +/- 22 min, respectively. At the point of correct displacement of the vocal cords, the average time from the end of propofol infusion until awakening with good quality of recovery was 313 +/- 93 sec. No patients complained of pain during or after CONCLUSIONS: This anesthetic technique provided both optimal operating conditions and patient comfort without serious complications. PMID- 17131892 TI - [Fentanyl shows different effects by administration routes on bispectral index during spinal anesthesia in patients undergoing cesarean section]. AB - BACKGROUND: Spinal anesthesia using a local anesthetic with fentanyl has been reported to induce sedation. We previously reported that the bispectral index (BIS) value was significantly decreased by spinal anesthesia using only bupivacaine and fetanyl after cesarean delivery. In the present study, we studied the effect of different fentanyl administration routes on BIS values during spinal anesthesia for cesarean section. METHODS: Forty-six women scheduled for cesarean section were allocated into five-groups according to the route of fentanyl administration and amount of local anesthetic: intrathecal 0.5% isobaric bupivacaine 2.5 ml plus fentanyl 20 microg (n = 11), intrathecal 0.5% isobaric bupivacaine 2.5 ml plus intravenous fentanyl 100 microg (n = 12), intrathecal 0.5% isobaric bupivacaine 2.5 ml plus epidural fentanyl 100 microg (n = 8), intrathecal 0.5% isobaric bupivacaine 2.5 ml (n = 8), and intrathecal 0.5% isobaric bupivacaine 3.0 ml (n = 7). BIS values were recorded during anesthesia. RESULTS: BIS values in intrathecal fentanyl group were lower than those of other groups (P = 0.03). The cumulative duration of BIS values 80 and below 80 was longer in the intrathecal fentanyl group than those of other groups (P = 0.004). CONCLUSION: The BIS value was significantly decreased only by intrathecal fentanyl for cesarean section. PMID- 17131893 TI - [Anesthetic management using vasopressin for a patient complicated with septic shock]. AB - We reported anesthetic management of a patient complicated with septic shock. Catecolamines were not effective to improve severe hypotension. Therefore, vasopressin was used and this improved the severe hypotension. Vasopressin may be useful for a patient complicated with septic shock and severe hypotension. PMID- 17131894 TI - [Bradycardia and cardiac asystole immediately after abdominal incision for removal of a huge pyometra]. AB - We report a patient who developed cardiac asystole, which may have been caused by Bezold-Jarisch reflex as a result of hypovolemia and compression of the inferior vena cava by a huge pyometra. A 61-year-old woman with a huge pyometra with occasional supine hypotension, tachycardia, and oliguria was scheduled for removal of the tumor. The systolic blood pressure decreased from 80 mmHg to 55 mmHg with simultaneous development of bradycardia 5 minutes after incision of the abdominal wall. Atropine was given but cardiac asystole occurred. Intravenous epinephrine restored systemic blood pressure and heart beats. There was no postoperative cardiorespiratory complication. PMID- 17131895 TI - [Aggravation of congestive heart failure after cesarean section in a patient with undiscovered hyperthyroidism]. AB - A 27-year-old woman with toxicity of pregnancy and congestive heart failure underwent an emergency cesarean section. In spite of antihypertensive drugs, blood pressure and heart rate were high preoperatively. She coughed many times and SpO2 was 88%. Circulatory state became stable without antihypertensive drugs under combined spinal-epidural anesthesia and continuous intravenous infusion of propofol intraoperatively. After surgery, blood pressure and heart rate became elevated again and congestive heart failure became worse. She was diagnosed as hyperthyroidism by postoperative laboratory examination. If we had discovered her hyperthyroidism, her symptoms of congestive heart failure would not have become worse. We should be more careful about hyperthyroidism in pregnancy because it is not rare. PMID- 17131896 TI - [Successful epidural anesthesia using 120 mm epidural needle for cesarean section in a morbidly obese parturient with body mass index 50.2 kg x m(-2)]. AB - In a morbidly obese parturient, epidural anesthesia is occasionally difficult because of great distance from the skin to the epidural space, and difficulty in identification of appropriate landmarks. We successfully managed cesarean section in a morbidly obese parturient with body mass index of 50.2 kg x m(-2) under epidural anesthesia. We used a 17 G custom-made epidural needle 120 mm long (Hakko, Tokyo, Japan), and the depth from the skin to the epidural space was 95 mm. We conclude that an extremely long epidural needle was useful in a morbidly obese parturient for overcoming the difficulties in epidural puncture and avoiding general anesthesia-related complications. PMID- 17131897 TI - [Acute popliteal arterial occlusion during extracorporeal circulation]. AB - A 79-year-old man underwent aortic arch replacement for thoracic aortic aneurysm. He had a history of smoking, coronary stenting for ischemic heart disease and replacement with artificial blood vessel for abdominal aortic aneurysm. Anesthesia was induced and maintained with midazolam, fentanyl, sevoflurane, and vecuronium. A 20 gage catheter was placed in the right radial artery and a 22 gage catheter in the left posterior tibial artery. Total circulatory arrest under profound hypothermia and retrograde cerebral perfusion were performed using extracorporeal circulation. After finishing anastomosis with artificial blood vessel, he was weaned from extracorporeal circulation. The pressure in the left posterior tibial artery was maintained at 15 mmHg, although the blood pressure in the right radial artery increased gradually. Then, the pressure in the left femoral artery in the operative field was the same as the pressure in the right radial artery. Therefore, we suspected the arterial line occlusion of the left posterior tibial artery. After the operation, we found the left leg and foot pale and cold with no pulsation on the left popliteal, dorsal pedis, and posterior tibial arteries. Further, acute left popliteal arterial occlusion was assessed by means of Doppler and left lower extremity angiography. We immediately performed the balloon-catheter embolectomy. However, as he developed compartment syndrome on the left lower limb due to reperfusion injury postoperatively, fascitomy was performed. On the 58th postoperative day, he was discharged from our hospital. Measurement by Doppler is useful for the early diagnosis of the lower leg arterial occlusion. PMID- 17131898 TI - [A case of craniofacial injury by javelin]. AB - We report a case of 16-year-old woman with craniofacial impalement injury by javelin (used by athletic sports) thrown from a distance of 30 meters. When the patient was admitted to the emergency department, the javelin had been in the orbita, and the patient was assessed by trauma team including an anesthesiologist. The patient was alert and complained of severe pain. Head CT and X-ray indicated the javelin had entered though the orbita into the maxillary sinus. Sphenoidal sinus was also partly destroyed by the impact. Anesthesia was soon induced in the emergency room to release the patient from the unbearable pain. To avoid tension cephalus, awake fiberscopic endtracheal intubation was performed. Anesthesia was maintained with propofol, vecuronium and fentanyl before transporting the patient to an operating room. The javelin and her head were strongly fixed with fiber cast to prevent from moving while she was transported to the operating room. The javelin was successfully removed during the operation. The fractures were fixed, and lacerations were sutured. The most important principle of management with craniofacial impalement injury is that the impaling object should remain in situ while patient is rapidly transported to an operating room, since it could extend tissue injury, and have a tamponade-like effect on damaged vascular structure. PMID- 17131899 TI - [Ventricular fibrillation during induction of anesthesia in a patient with polycythemia vera]. AB - A 53-year-old male patient complained of the pain with bilateral hip area and right hip joint and underwent emergency arthroscopy and drainage. Twenty-eight years before, he had suffered from gout and from his abnormal increase of blood cells was diagnosed as polycythemia vera. The laboratory examination at admission showed a marked increase of hemoglobin (17.7 g x dl(-1)) and hematocrit (69.5%). Immediately before induction of anesthesia, 1000 ml of phlebotomy was performed with large fluid infusion. After induction of anesthesia and oro-tracheal intubation, electrocardiogram (ECG) suddenly showed ventricular fibrillation (Vf). Defibrillation was applied and the ECG recovered to sinus rhythm, but 30 min later, ECG showed Vf, again. The increased blood viscosity with polycythemia might have induced coronary ischemia and fatal arrhythmia in the patient. Vigourous hemodilution before surgery should have been performed as prophylactic management of the cardiac episodes in this patient. PMID- 17131900 TI - [OSCE improves medical student's BLS knowledge, but its long-term memory is not well maintained]. AB - BACKGROUND: Objective structured clinical examination: OSCE is now widely used in educational scene in medical schools before medical students start their clinical training. However, the utility of OSCE on BLS education has not been confirmed yet. The effect of OSCE on BLS education was examined using questionnaires. METHODS: Questionnaires on knowledge of resuscitation were distributed to 166 medical school students (100 students in 1999 semester without OSCE, 66 in 2004 semester with OSCE). The questionnaire included 13 questions based on the 1992 or 2000 guidelines for cardiopulmonary resuscitation. Six out of 12 questions were defined as "skill related questions" and the student who correctly answered all these "skill related questions" were thought to be competent for providing adequate BLS. RESULTS: Competent student ratio was significantly higher in students with OSCE (48%, vs 20% without OSCE, P < 0.001). Thus OSCE was useful for medical student to establish BLS knowledge when compared with students without OSCE. However, this result also revealed that the acquired knowledge through OSCE did not last long. Although 98% of students actually passed OSCE, only half of them were competent on paper test during clinical training. CONCLUSIONS: OSCE improved medical students' knowledge of BLS, but did not contribute to prevent forgetting for a long time. Thus, reeducation in BLS is still necessary for students after passing OSCE. PMID- 17131901 TI - Challenges in ambulatory resident education: medication knowledge in disadvantaged patients. AB - CONTEXT: Medication mismanagement results in 10% of hospitalizations and 50% of therapeutic failures. Little is known about low-income patients' medication knowledge, a group disproportionately affected by chronic disease and often cared for by residents. This study assesses patients' medication knowledge, discrepancies between the patient and chart, and preferences for medication information in the training setting. DESIGN: This cross-sectional study evaluated adults receiving care in two teaching clinics. Patients were asked their medications' names, doses, frequencies, indications, and side effects (SE) and what information they needed to take medications safely. Charts served as a medication reference standard. RESULTS: Most patients were female, nonwhite, had > or = one chronic condition, took a median of five drugs, and saw three different residents in the same clinic. Despite six clinic visits annually, patients could cite only 50% of their medication names, frequencies, and indications, 25% of doses, and 0% of SE, yet they paradoxically responded that SE knowledge was essential for safety. Forty-three percent reported > or = one medication that was undocumented in the chart. There were 84 total discrepant medications of which 63% were prescription only. CONCLUSION: Low-income patients had poor medication knowledge despite good clinic access. Discontinuity of resident providers may contribute to this phenomenon. Poor global knowledge, the large discrepancy between reported and charted medications, and lack of SE knowledge threaten medication adherence, achievement of treatment goals, and patient safety. PMID- 17131902 TI - A New England bonesetter dynasty: the Porters of Farmington, Connecticut--Part I of II. AB - Daniel Porter (d. 1690), a bonesetter/ chirurgeon, one of the early settlers of the Connecticut Colony, was the first and only doctor serving the Farmington valley during much of the second half of the 17th century. More than 40 of his descendants, spanning eight generations and almost three centuries, practiced medicine and surgery, moving with the migration of the expanding population to towns throughout New England, New York State, and ultimately the midwest and west coast. Their notable accomplishments, apart from their consistent reputation for exceptional skill and endurance, include the training and independent practice as bonesetters of two of their wives. In 1796, Samuel Lee, Jr., (1773-1814), a fifth generation descendant, received the first U.S. patent for a medication, "Samuel Lee's Genuine Windham Bilious Pills." The last Porter doctor to be identified, Charles Allen Porter (1866-1931), a pioneer Professor of Surgery at Harvard Medical School, was among the first to recognize the carcinogenic effects of radiation. In the absence of a comprehensive published genealogy of the Daniel Porter family, this essay identifies the relationships among his many descendants. PMID- 17131903 TI - Credentialing oversight equals hospital negligence? PMID- 17131904 TI - Some problems of medical education. 1956. PMID- 17131905 TI - Mind and brain: together again. PMID- 17131906 TI - Suggested additions to the patient information booklet--prior to colonoscopy. PMID- 17131907 TI - Is the anti-sarcoma and anti-viral cytokine "plasma factor" a novel chicken Y-box protein? AB - A line of research beginning in the early 1960s with the observation that West Nile virus and, later, several strains of rabies virus could inhibit the development of the Rous sarcoma virus-induced tumor in the wing-web of chicken (a "sarcoma-blockade") eventually culminated in the characterization of a 14-kDa circulating anti-sarcoma and anti-viral activity christened "plasma factor" (PF) which, unlike the interferons, inhibited the replication of diverse RNA containing viruses, but not of any DNA-containing viruses. The possibility that this 14 kDa protein represented a novel antiviral cytokine has been strengthened by analysis of partial amino acid sequencing data which suggest that this 14-kDa cytokine may correspond to the 127-amino acid-long chicken YB2-like protein (Locus: XP_423576) deduced very recently from the genomic sequencing of chicken. Biologically, proteins of the Y-box family (such as chicken YB1 and YB2) not only bind DNA and thus regulate transcription but also bind single-stranded RNA in a sequence-specific and reversible manner, repress viral RNA translation, inhibit retroviral transformation of chicken fibroblasts, and are known to regulate transcription of human immunodeficiency virus and hepatitis B virus. Taken together, the available data point to a novel anti-viral cytokine with a novel mechanism of action. PMID- 17131908 TI - Binding characteristics and distribution of lactoferrin receptors in the gut and choroid plexus in newborn calves. AB - Lactoferrin (Lf), an iron-binding multifunctional glycoprotein, abundantly present in colostrum and milk of different species such as humans, bovines, and mice has been shown that bovine colostral Lf is transported into the CSF via plasma in newborn calves. Specific Lf-receptors (Lf-R) are present in different cells of different species. In the present study, we report for the first time, the presence and distribution of Lf-R in the intestine and choroid plexus in newborn calves. Brush-border membrane vesicles (BBMV) were prepared from the mucosa of duodenum, jejunum, ileum, colon, epithelium overlying Peyer's patches (EOPP) in jejunum (EOPPJ) and ileum (EOPPI), and choroid plexus membranes. Receptor binding assays were carried out using 125I labeled bovine Lf. Specific and saturable Lf-R were found in BBMV of all the intestinal segments and choroid plexus examined. Nonlinear regression and Scatchard plot analyses clearly revealed that EOPP had the highest binding maximal (Bmax), and lowest in colon. The maximum dissociation constant (Kd) 0.7 microM was in colon while, Bmax and Kd in choroid plexus membrane were 16.87 nmol/mg protein and 0.34 microM, respectively. All these findings together strongly suggested that Lf was transported into CSF via plasma through receptor mediated transcytosis. PMID- 17131909 TI - Modulation of lecithin activity by vitamin-B complex to treat long term consumption of ethanol induced oxidative stress in liver. AB - Alcoholic liver disease (ALD) develops as a consequence of priming and sensitizing mechanisms rendered by cross-interactions of primary mechanistic factors and secondary risk factors. Chronic alcohol abuse and its progression to ALD are associated with abnormal metabolism and low tissue or plasma levels, or both, of many micronutrients. Glutathione depletion is considered the most important sensitizing mechanism. In the present study efficacy of lecithin with vitamin-B complex to treat ethanol induced oxidative stress was compared with the effect of lecithin alone, tocopheryl acetate (vitamin E), as well as capacity of hepatic regeneration during abstention. Ethanol (1.6g / kg body weight/ day for 4 weeks) affects body weight in 16-18 week old male albino rats of Wistar strain weighing 200-220 g. Thiobarbituric acid reactive substance level, nitrite content, protein carbonyl group level, redox ratio (oxidized to reduced glutathione ratio), superoxide dismutase activity, and glutathione s-transferase activity significantly increased on ethanol exposure. Whereas reduced glutathione content, and activities of catalase, glutathione reductase and glutathione peroxidase significantly reduced due to ethanol exposure. These changes were reversed by different treatment. The results suggest that tocopheryl acetate (vitamin E) could partially reverse these changes and act as a potential therapeutic agent. However, lecithin with vitamin-B complex treatment is a promising therapeutic approach. Furthermore, preventive measures were more effective than curative treatment. Prevention of oxidative and nitrosative stress along with correction of nutritional deficiency is one of the proposed mechanisms for the therapeutic approach. PMID- 17131910 TI - Heat stress induced apoptosis in BC-8 cells derived from AK-5 tumor involves downregulation of Bcl-2 and generation of reactive oxygen species. AB - BC-8, a rat histiocytoma undergoes apoptosis after heat shock, which is due to lack of an effective heat shock response. Heat shock induced generation of free radicals, which in turn are involved in the induction of apoptotic death in BC-8 cells. Treatment of BC-8 cells with N-acetylcysteine partially inhibited the heat induced apoptosis. Introduction of Bcl-2 gene in these cells did not protect them from apoptotic death, whereas transfection with hsp-70 gene did render these cells resistant to heat induced apoptosis transiently. Heat shock also downregulated the expression of Bcl-2 and p53 in these cells. These observations suggested that the heat shock induced apoptosis was mediated through reactive oxygen species and controlled upstream of Bcl-2 check point. PMID- 17131911 TI - Involvement of nitric oxide (NO) in the regulation of stress susceptibility and adaptation in rats. AB - The present study evaluated the regulatory role of nitric oxide (NO) in stress susceptibility and adaptation in rats. Acute restraint stress (RS x1) reduced the number of entries and time spent in the open arms in the elevated plus maze (EPM) test and raised plasma corticosterone levels. RS (x1)-induced neurobehavioral suppression and raised corticosterone levels were attenuated by pretreatment with the NO precursor, L-arginine (500 and 1000 mg/kg)and unaffected or further aggravated by NO synthase inhibitor, L-NAME or 7-nitroindazole (10 and 50 mg/kg). Biochemical assay of plasma and brain homogenates showed that these RS - induced behavioral and neuroendocrinal changes were associated with lowered levels of plasma and brain total nitrates/nitrites (NOx). L-Arginine attenuated the RS induced suppression of NOx levels in plasma and brain, whereas, the NO synthase inhibitors tended to produce reverse effects. In the experiments involving repeated stress i.e. RS (x5), exposure resulted in attenuation/reversal of (a) neurobehavioral suppression in the EPM test and (b) lowered brain NOx, that was seen after RS (x1). The RS (x5)-induced changes in EPM parameters and brain Nox were further potentiated after L-arginine pretreatment, whereas, the NO synthase inhibitors were less effective. Rats were screened as high and low emotional in the open-field test, and high emotional rats showed greater(a) behavioral suppression in the EPM, (b) corticosterone responses (c) brain NOx suppression, and (d) cold-restraint stress (CRS) induced gastric mucosal lesions as compared to their low emotional counterparts. L-Arginine pretreatment was more effective in modulating the above RS induced stress responses/markers in the high emotional group of rats. Our data suggest that NO plays a differential role during exposure to acute and repeated stress situations, and that the relationship between stress and emotionality status may be under the regulatory influence of NO. PMID- 17131912 TI - Role of free radicals in stress-induced neurobehavioural changes in rats. AB - Effect of restraint stress (RS) and its modulation by antioxidants were evaluated on elevated plus maze (EPM) and open field (OF) tests in rats. Restraint stress (RS for 1 hr) reduced the number of open arm entries, as also the time spent on open arms indicating enhanced anxiogenic response in the EPM test as compared to normal non RS group of rats. Pretreatment with ascorbic acid (100 and 200 mg/kg) and alpha-tocopherol (30 and 60 mg/kg) attenuated these RS-induced effects. In the OF test, RS-reduced (a) ambulations; and (b) rearings, whereas an increase was seen in (a) latency of entry and (b) number of fecal boluses. The RS-induced changes in OF parameters were reversed after pretreatment with the antioxidants, (ascorbic acid and alpha tocopherol). Biochemical data showed that RS enhanced MDA levels in both serum and brain, and these were attenuated after pretreatment with the antioxidants. The pharmacological and biochemical results indicate that free radicals might be involved in such stress-induced neurobehavioural effects. PMID- 17131913 TI - Protective effect of ethanolic and water extracts of sea buckthorn (Hippophae rhamnoides L.) against the toxic effects of mustard gas. AB - Ethanolic extract of H. rhamnoides L. leaf (HL-EOH), water and ethanolic extract of H. rhamnoides fruit (HF-W and HF-EOH), and H. rhamnoides flavone from fruit (HR-flavone) were evaluated against percutaneously administered sulphur mustard (SM), a chemical warfare agent. The animals administered with SM (9.7, 19.3 and 38.7 mg/kg) died at various days depending upon the dose and there was a significant reduction in the body weight. The H. rhamnoides extracts (1 g/kg; 3 doses; po) significantly protected the lethality, with a protective index of 2.4, 1.7, 1.7 and 2.2 for HL-EOH, HF-W, HF-EOH and HR-flavone respectively. Reduced glutathione (GSH) and oxidized glutalthione (GSSG) levels were reduced, and malondialdehyde (MDA) was elevated after percutaneous administration of SM. Oral administration of HL-EOH and HR-flavone significantly protected the body weight loss. Recovery in the levels of GSH, GSSG and MDA were also observed following oral administration of HL-EOH and HR-flavone. All the extracts were non-toxic and the LD50 was more than 5 g/kg. The present study shows that percutaneous administration of SM induces oxidative stress and ethanolic extract of leaf of H. rhamnoides and H. rhamnoides flavone from fruit can significantly protect it. PMID- 17131914 TI - Antioxidant property of Decalepis hamiltonii Wight & Arn. AB - Aromatic edible root of D. hamiltonii was subjected to the extraction of the antioxidant rich fraction. Different parts of root namely whole tuber, peel, tuber without peel and medullary portion were extracted with dichloromethane (European Patent No. W02005063272). The extract was found to contain flavor compound 2-hydroxy-4-methoxybenzaldehyde (2H4MB), which was identified by TLC and GC. Medullary portion was found to be rich in 2H4MB, (73.73 mg g(-1) dry tissue) followed by peel, containing 68.34 mg g(-1) 2H4MB. Different concentration of dichloromethane extracts were subjected for antioxidant assay by DPPH (1,1 dihydroxy 2-picryl hydrazyl) method, this has shown 44, 46.7% radical scavenging activity in case of medullary, peel extracts and 67.3% in case of pure 2-hydroxy 4-methoxybenzaldehyde at 100 ppm concentration, whereas ascorbic acid used as standard showed 94.3% activity. In beta-carotene linoleate model system (b-CLAMS) 43.46 and 45.7% antioxidant activity was observed in medullary and peel extracts at 100 ppm concentrations respectively, whereas standard 2-hydroxy-4 methoxybenzaldehyde exhibited 69.64% at 100 ppm and BHA (butylated hydroxyl anisole) 90.1% activity also at 100-ppm level. Similarly hydroxyl radical scavenging activity was found to be 48.36, 46.86, 48.26 and 73.60% in whole tuber, medullary, peel and standard 2-hydroxy-4-methoxy benzaldehyde respectively at 100 ppm levels. This is the first report on the antioxidant activity of D. hamiltonii. Results have shown that 2H4MB is one of the major constituents responsible for antioxidant activity. Hence the extract of D. hamiltonii can be utilized for the production of antioxidant rich fractions required for various health benefits. PMID- 17131915 TI - Ginseng extract exhibits antimutagenic activity against induced mutagenesis in various strains of Salmonella typhimurium. AB - Ginseng has been reported to exhibit antioxidant and antimutagenic activity. The present study was undertaken with a view to confirm whether the antioxidant activity of Ginseng is responsible for its antimutagenic action. The concentrated root extract of Panax ginseng (Ginseng extract I) and its lyophilized powder (Ginseng extract II) obtained from two different manufacturing houses, were tested against mutagenesis using the well-standardized Ames microsomal test system. The extracts exhibited antimutagenic effect against hydrogen peroxide induced mutagenesis in TA100 strain, and against mutagenesis produced by 4 nitroquinoline-N-oxide in both TA98 and TA100 strains of Salmonella typhimurium. Both the extracts failed to show any antimutagenic potential against tert-butyl hydroperoxide (an oxidative mutagen) in TA102 strain, a strain highly sensitive to active oxygen species. The extracts also indicated a weak antioxidant activity in a series of in vitro test systems viz., 1,1-diphenyl picryl hydrazyl (DPPH) assay, hydrogen peroxide scavenging and superoxide anion scavenging. The results indicate that the protective effects shown by ginseng extract(s) against 4 nitroquinoline-n-oxide and hydrogen peroxide induced mutagenesis in TA98 and TA100 could mainly be due to its property to initiate and promote DNA repair rather than free radical scavenging action. PMID- 17131916 TI - Optimization of nutritional requirements for gentamicin production by Micromonospora echinospora. AB - Effect of various fermentation media, carbon sources, nitrogen sources, phosphate concentration and culture requirements includes inoculum levels and age were determined on gentamicin production and biomass dry weight production for Micromonospora echinospora, a gentamicin producing strain. Of the substrates tested, starch as a sole carbon source promoted maximal gentamicin production, while maltose promoted maximal growth. Yeast extract as a sole nitrogen source promoted maximal growth, while soyabean meal for gentamicin production. Increasing phosphate concentration enhanced gentamicin production and observed optimum production at 1.2 g/1 (6% v/v) of phosphate having 72 h old inoculum in the medium. Highest gentamicin production was obtained after cultivation with shaking for 120 h in a medium containing starch 0.75% (w/v), soyabean meal 0.5%, K2HPO4 0.12%, CaCO3 0.4%, FeSO4 0.003% and CoCl2 0.0001%. The gentamicin production was 1.2-fold in this medium as compared to basal medium. PMID- 17131917 TI - Iron induced metabolic changes in the diazotrophic cyanobacterium Anabaena PCC 7120. AB - Iron induced changes in growth, N2-fixation, CO2 fixation and photosynthetic activity were studied in a diazotrophic cyanobacterium Anabaena PCC 7120. Iron at 50 microM concentration supported the maximum growth, heterocyst frequency, CO2 fixation, photosystem I (PS I), photosystem II (PS II) and nitrogenase activities in the organism. Higher concentration of iron inhibited these processes. Chl a and PS II activities were more sensitive to iron than the protein and PS I activity. PMID- 17131918 TI - DNA isolation from goat blood using different brands of household detergents and its downstream application. AB - Rapid isolation of DNA from goat blood using different brands of detergents available in Indian market, is reported. The integrity and efficiency of these DNA preparations were compared with genomic DNA isolated by a standard kit (Flexi gene DNA kit), using amplification of exon 2 of CYP19 (aromatase) gene. The similar and significant amplification of this gene was obtained using genomic DNA isolated by kit and various detergents. However, among the detergents used, the Rin and Ezee were found to be the best to get DNA of high purity comparable to that obtained by kit. PMID- 17131919 TI - [OSCAR (one-stop clinic for assessment of fetal risk): our experience with first trimester screening for chromosomal abnormalities]. AB - OBJECTIVES: In recent years has been intensified an effort to develop early more exact non-invasive methods for prenatal diagnosis. We report our experience to screen trisomy 21 and other chromosomal abnormalities in the first trimester in so-called OSCAR (One-Stop Clinic for Assessment of Fetal Risk). DESIGN AND SETTING: It is a retrospective study held at the Department of Medical Genetic and Fetal Medicine, Teaching Hospital, Olomouc. METHODS: A group of 2110 pregnant women has been screened by maternal age as a background risk, fetal nuchal translucency, nasal bone as ultrasound markers as well as maternal serum level of free beta-subunit human chorionic gonadotropin (beta-hCG) and pregnancy associated plasma protein-A (PAPP-A) as a biochemical markers between 11 - 13+6 weeks of pregnancy from January 2004 to December 2005. The Fetal Medicine Foundation Guidelines and software were the methods and tools used for screening. Karyotyping by chorionic villous sampling or amniocentesis was offered to women with risks > or = 1 in 300. RESULTS: We reported 100% sensitivity for this method for a 4, 6% false positive. There was detected 21 chromosomal abnormalities in which 10 were trisomy 21. This is the first result to be published for this screening method in the Czech Republic. CONCLUSION: In our experience first trimester screening for trisomy 21 and other aneuploidies has a high sensitivity with a low false positive rate and can be delivered in an efficient manner in a one-stop multidisciplinary clinic. PMID- 17131920 TI - [Insertion-deletion polymorphism in the gene for angiotensin-converning enzyme (I/D ACE) in pregnant women with gestational diabetes]. AB - AIM OF THE STUDY: The aim of the study was to investigate possible associations of the Angiotensin-converting enzyme insertion/deletion (I/D ACE) polymorphism with gestational diabetes mellitus occurrence. METHODS: Number of 53 healthy pregnant women (controls) and 48 women with gestational diabetes mellitus were included in the study. The woman patients were hospitalised in the Clinic of Obstetrics and Gynaecology, Masaryk University Affiliated hospital Brno during the period 10/2004 - 10/2005. In all cases there was the spontaneous one baby pregnancy. The procedure was obtaining the peripheral blood samples from women of those two groups and isolation of their DNA using standard technique with the use of proteinase K. I/D ACE genotype of each person was determined using allele specific primers. RESULTS: We did not prove significant association neither of the genotype distribution (P(g)) P(g) = 0.115 nor allele frequency (P(a)) P(a) = 0.873 between controls and women with gestational diabetes mellitus. The significant differences of genotype distributions (P(g) = 0.03) were observed in women with two and more labours in anamnesis. Borderline significant differences of genotype distributions (P(g) = 0.08) were found in women with one and more abortions in anamnesis. CONCLUSIONS: According to our results the variability in ACE gene can be taken as factor contributing to manifestation of gestational diabetes mellitus. According to results of our study I/D ACE polymorphism can't be taken as genetic marker of gestational diabetes mellitus. PMID- 17131921 TI - [A role of prenatal diagnostics in birth defects occurrence in the Czech Republic in 2004]. AB - OBJECTIVE: An analysis of prenatal diagnostics of birth defects in the Czech Republic in 2004 and a comparison to data from 1996 - 2003. TYPE OF STUDY: A retrospective analysis of data from a Register of Prenatal Diagnostics of the Czech Republic. METHODOLOGY: Data on birth defects in the Czech Republic from particular departments of medical genetics and prenatal diagnostics from 1996 to 2004 time period was used. An analysis of data on prenatal diagnostics of selected type of birth defects was performed. RESULTS: During 1996 to 2004 period, a total of 4,126 prenatally diagnosed and terminated cases were reported. In 2004, 597 cases were reported, which presents 61.1 cases per 10 000 live births. (The mean rate from the total period under the study was 49.8 per 10 000 live births.) CONCLUSIONS: Current data on birth defects in a 9-year period in the Czech Republic are presented. The role of prenatal diagnostics in decrease of some severe types of birth defects is underlined. PMID- 17131922 TI - [Infant mortality due to birth defects in the Czech Republic in 1994-2004]. AB - AIM AND TYPE OF STUDY: A retrospective data study with an analysis of pre- and postnatal occurrence of selected types of birth defects in the Czech Republic during 1994 to 2004 period. An analysis of infant mortality rate in children with selected types of birth defects during their first year of life is described. METHODOLOGY: Data on prenatal diagnostics were obtained from particular departments of medical genetics. Data on birth defects incidences were obtained from National Health registers - Institute of Health Information and Statistics (National Register of Congenital Anomalies and National Newborns Register) from the 1996-2004 period were used. A case analysis of incidences of 14 types of pre- and postnatally diagnosed birth defects was performed for the period 1994-2004. In postnatally diagnosed cases, an analysis of survival and extinction of particular types of defects during the first year of life was also performed. RESULTS: During 1994-2004, there were 1 030 069 children born in the Czech Republic. Including prenatally diagnosed cases, we registered 305 cases of anencephaly, 454 spina bifida, 156 encephalocele, 528 cases of congenital hydrocephalus, 304 omphalocele and 302 cases of gastroschisis, 241 oesophageal defects, 302 anorectal malformations, 312 cases of diaphragmatic hernia, 518 renal agenesis/hypoplasia cases, 529 cases of cystic kidneys and 1764 Down syndrome cases. Secondary prevention rate was high in neural tube defects (61 100%), abdominal wall defects (60-79%) and Down syndrome (63%). In operable defects, a first-year survival was lowest in congenital hydrocephalus (63%) and neural tube defects -excluding anencephaly - (82%). In abdominal wall defects, diaphragmatic hernia, oesophageal and anorectal malformations, the survival rate was, however, 76-91% of children. CONCLUSIONS: Birth defects present a major contribution to infant mortality and morbidity. Prematurity in a combination with a birth defect prolongs inpatient days and increases a need of a total parenteral feeding in a newborn and it also makes a worse prognosis for the infants with birth defects. An improvement of prenatal diagnostics with an early detection of the most severe types along with the associate ones could decrease incidences of such cases. A progress in surgical techniques and a special neonatal and infant intensive health care would also contribute an improvement of survival. PMID- 17131923 TI - [Pregnancy after laparoscopic myomectomy--long-term follow up]. AB - OBJECTIVE: Analysis of pregnancy related complications in infertile patient after laparoscopic myometomy (LM). DESIGN: Retrospective analytic study. SETTING: Institute for the Care of Mother and Child, Prague; Institute for Postgraduate Medical Education, Prague. METHODS: The study group were infertile patients referred to our department with the finding of uterine fibroid(s). In all patients LM was the method of choice. Thorough information and results from all infertile patient after LM were assessed and analysed with 18 months follow-up after surgery. RESULTS: The aim of our study was to evaluate pregnancy rate and the possible risks during pregnancy and delivery following LM. We analysed 69 patients after LM. Mean size of a fibroid was 3.3 cm and the number of removed fibroids in one patient was 1.8. The conception rate after LM was 56.5%. We didn't observe any increased incidence of fetomaternal morbidity or severe pregnancy and labour related complications. There was no uterine rupture after LM in our group. The cesarean section was rate 44.8%. CONCLUSIONS: The impact of fiborids on infertility is still controversial. LM in infertile patient is one of the most common surgical procedure. The appropriate surgical management of uterine scar is mandatory. This operation must be performed by skilled reproductive surgeon. Thorough information to the patient before and after LM is necessary. The pregnancy following LM is at high risk with increased caesarean section rate. PMID- 17131924 TI - [Preoperative assessment of myometrial invasion in endometrial cancer patients by ultrasonography and magnetic resonance imaging (MRI)]. AB - OBJECTIVE: To compare the accuracy of transvaginal sonography and magnetic rezonance imaging (MRI) in determining the depth of myometrial invasion in proven cases of endometrial cancer. DESIGN: A prospective study. SETTING: Department of Obstetrics and Gynecology, Department of Medical Genetics and Fetal Medicine, Department of Radiology, University Hospital, Olomouc. METHODS: Fifty five patients diagnosed as having FIGO stage I endometrial carcinoma were evaluated preoperatively by transvaginal ultrasound; 44 cases of the same group were also evaluated by magnetic resonance imaging (MRI). The degree of invasion evaluated by transvaginal sonography and by MRI was compared to the pathological specimens. RESULTS: Transvaginal sonography was successful in evaluating myometrial invasion in 44 of 55 cases (accuracy 80%, sensitivity 80%, specificity 88%, positive predictive value 77%, negative predictive value 87%). Evaluation with MRI was accurate in 37 of 44 cases (accuracy 84%, sensitivity 84%, specificity 91%, positive predictive value 81%, negative predictive value 91%). CONCLUSION: Although MRI is superior to transvaginal sonography in evaluating myometrial invasion, it is expensive and time consuming, and would not be suitable as a screening test for depth of invasion. On the other hand, transvaginal sonography is relatively low-cost technique, which can be easily performed and repeated. However, it requires more operator experience than MRI in order to achieve high accuracy. PMID- 17131925 TI - [Possibilities of using ultrasound in the diagnosis of descended posterior compartment of female pelvic floor]. AB - OBJECTIVE: Find out the features of descending posterior vaginal wall using ultrasonography and set the objective diagnostic criteria. DESIGN: Prospective comparative study. SETTING: Department of Obstetrics and Gynaecology, Teaching Hospital Bulovka, First Medical Faculty, Charles University in Prague. METHODS: We included 39 attendants, 19 with clinicaly proven descent of posterior vaginal wall; 20 as a negative control group. We observed the ultrasonographical features of descending posterior vaginal wall according to the horisontal line crossing the inferior margin of pubic bone (PM) and central anorectal angle (PARA) at rest and during Valsalva manoevre with and without intrarectal application of sonographic yelly. Student's t-Test was used for statistical evaluation. RESULTS: We proved the statisticaly significant increase in the distances PM and PM' in the group of females suffering from the descent compared to the group of healthy women. Values of PARA were also signifinatly hightened in the group of patients with the descent compared to healthy females. CONCLUSIONS: The ultrasonographical evaluation of descending posterior vaginal wall appears to be promising chance in diagnostics of female's pelvic floor pathology. PMID- 17131926 TI - [Delayed interval delivery in twins pregnancy--the case report]. AB - OBJECTIVE: The autors demonstrate the case report of the 70 days delayed interval delivery in twins pregnancy in 24 g.w. DESIGN: Case report. SETTING: Department of Gyneacology and Obstetrics Most. METHOD: 31-years old primipara after IVF pregnancy with preterm rupture of membranes of fetus A, after induced uterine contractions terminated the abortion of fetus A. The contractions stopped with tokolysis and after one month of expectation interval was performed cerclage suture of cervix. CONCLUSION: After the abortion of twin A, we succesfully delayed interval delivery for 70 days. The borderline mature neonate was born in good clinical condition and survive. PMID- 17131927 TI - Transvaginal hydrolaparoscopy (THL) in diagnosing infertility. AB - INTRODUCTION: In recent years novel gynecological diagnostic methods have emerged. One of them is transvaginal hydrolaparoscopy (THL). This method allows endoscopic visualization of the organs within the women pelvis accessed through the posterior fornix of the vagina (Veress's needle was used) and with peritoneal distension by saline. OBJECTIVE: To evaluate a THL as a new technique for diagnosing pelvic pathology. METHODS AND RESULTS: In years 2002-2004, 56 patients underwent THL due to primary (40 cases) or secondary (16 cases) infertility (age 20-40). None of the patients had any history of previous pelvic surgery and all had normal findings on gynecological examination and vaginal sonography. The quality of the pelvic pictures was satisfactory in all cases. In the infertile group we observed in 42.8% cases of no macroscopic changes, in 57.2% some form of pelvic pathology ; 28.6% of demonstrated endometriosis, and 28.6% had tubal occlusion. There were no complications during or after the operation. CONCLUSIONS: THL is well-tolerated and accepted by the patients. THL is a safe, simple and inexpensive diagnostic method for pelvic pathology and infertility. PMID- 17131928 TI - [Lymphatic mapping and detection of sentinel node in early stages of cervical carcinoma]. AB - OBJECTIVE: The aim of the study was to analyse two methods of intraoperative sentinel node detection using blue dye and blue dye with Tc99 labeled tracer in early stage of the cervical cancer. DESIGN: Prospective case observational study. SETTING: Department of Obstetrics and Gynecology, University Hospital Ostrava. METHODS: From May 2004 to September 2005, 49 patients with cervical cancer who underwent a radical surgery were included into the study. Sentinel lymph node was detected using blue dye in the first group of 23 patients and by blue dye with Tc99 in the second group of 26 patients. Intraoperative sentinel node detection was performed by visual aspection in the first group, and by visual aspection and by hand-held gamma-probe in the second group. RESULTS: Patients were divided according to stage of the disease into three subgroups FIGO IA2, FIGO IB1 and FIGO IB2. A total number of 1561 lymph node with an average 32 and 94 SLN with an average 1.9 were identified. The specific detection rate per site was 63% in the first group and 80.8% in the second group respectively. Metastatic disease was detected in 26 patients (41%) and metastatic involvement of SLN only in 11 patients (17.4%). Sensitivity and negative predictive value were 100% in both groups, false negativity was 0%. CONCLUSION: Detection of SLN by combination of blue dye and Tc99 labeled tracer has a higher detection rate of SLN than detection by blue dye alone. PMID- 17131929 TI - [Effectivity and complications rate of surgical treatment of stress urinary incontinence with tension-free vaginal tape--Uretex]. AB - OBJECTIVE: To evaluate the safety and efficacy of the Uretex tension-free vaginal tape procedure in the treatment of female stress urinary incontinence. DESIGN: Retrospective multicenter clinical trial. SETTING: Obstetrics and Gynecology Department, The Charles University 2nd Medical School and Teaching Hospital Motol, Prague. METHODS: A group of 145 patients with stress urinary incontinence who underwent Uretex tension-free vaginal tape procedure was studied. Procedures were performed from vaginal retropubic route. Patients were operated on in one urological and five gynecological departments in Czech Republic. Preoperative evaluation consisted of complete urogynecological examination, age, parity, body mass index, history of previous pelvic surgery and hormonal status. The cure rate and complications were determined in short-term postoperative follow-up. RESULTS: The mean age was 56 (25-80) years, the median parity was 2.25 (1-4), 113 (78%) patients were slightly overweight (BMI 25-30 kg/m2) and others had normal weight (BMI 20-25 kg/m2). The cure rate after surgery was 903% (131 women). Complications occured in 22 (15.2%) cases. In two (1.4%) patients perforation of the urinary bladder wall occurred. In two (1.4%) women nonperforated injury of bladder wall was postoperatively diagnosed. Retropubic haematoma occurred in another two patients (1.4%). 16 (11%) patients had mild early postoperative complications (eight times short-term urinary retention, six lower urinary tract infections, two urgent symptomatology). Erosion of vaginal wall did not occur postoperatively. All complications were resolved and the patients were free of negative postoperative symptoms 6 month after the procedure. CONCLUSION: This study shows that Uretex tension-free vaginal tape procedure is an effective and safe minimally invasive surgical procedure in the treatment of stress urinary incontinence. PMID- 17131930 TI - [Fetal expression by squeezing the uterine fundus]. PMID- 17131931 TI - [Guideline recommendations for gestagen contraception. 2006]. PMID- 17131932 TI - [Recommendations for diagnosis and therapy of premenstrual syndrome (PMS). 2006]. PMID- 17131933 TI - The role of cytokines in the immune response to influenza A virus infection. AB - Influenza A virus is one of the most important causes of respiratory tract diseases. It replicates in epithelial cells and leukocytes resulting in the production of immune mediators--cytokines, substances with various biological effects. Cytokines, as a part of innate immunity, favor the development of antiviral and TH 1-type immune responses. Cytokines also affect the adaptive immune response and disease manifestation. In the organism, the virus infection results in the production of chemotactic [a regulated upon activation, normal T cell-expressed and -secreted cytokine (RANTES), monocyte chemoattractant proteins (MCP) MCP-1, MCP-3, macrophage inflammatory protein 1 alpha (MIP- 1 alpha), interferon gamma-induced protein 10 (IP-10), and interleukin 8 (IL-8)], pro inflammatory [IL- 1beta, IL-6, IL-18, and tumor necrosis factor alpha(TNF-alpha)] and antiviral [interferon (IFN) alpha/beta] cytokines. Whilst knowledge of the mechanisms underlying host and tissue specificity has advanced significantly, we still know relatively little about the function of cytokines released from different cells following influenza infection. In this review we deal with the role and mode of possible impact of cytokines on the disease pathogenesis and host immune response. PMID- 17131934 TI - Biological and molecular characterization of Avian influenza virus (H9N2) isolates from Iran. AB - Three Influenza A virus (H9N2) isolates obtained from three separate broiler flocks with variable mortality rates were cloned twice in embryonated SPF chicken eggs by limiting dilution. Biological properties of these isolates were examined in 4-week-old SPF chickens and chick embryo fibroblast (CEF) cultures. The isolates neither caused mortality in the inoculated chickens nor produced CPE in cell cultures, indicating low pathogenicity. PCR products of 486 bp containing the sequences for hemagglutinin (HA) cleavage site, which were generated from the isolates, were subjected to nucleotide sequencing. Sequence analysis of the HA region containing the cleavage site of the isolates showed a similar sequence motif (PARSSRG) but different flanking regions. Phylogenetic analysis of deduced amino acid sequences revealed that the isolates were closely related to those isolated earlier, indicating a common source. Moreover. the amino acid sequences of the recent isolates were very similar to those from Saudi Arabia, Germany and Pakistan. It is postulated that, except for some Chinese isolates, the pathogenicity of Iranian isolates seems to be similar to that of other Eurasian isolates. It is possible that an elevation in mortality rate under field condition could be caused by co-infection of recent isolates with the bacteria such as mycoplasma, Escherichia coli, and Ornithobacterium rhinotracheale rather than by an emerging a pathogenic H9N2 subtype of the virus. PMID- 17131935 TI - Formation of Pseudorabies virus glycoprotein E/I complex in baculovirus recombinant system. AB - Glycoproteins E (gE) and I (gI) of Pseudorabies virus (PRV) form a non-covalently bound complex to which a number of functions have been attributed. The gE/gI complex formation was studied using a series of full-length and truncated forms of gE and gI expressed in baculovirus recombinant system. Both glycoproteins were truncated by stepwise removal of their C-terminal parts and their ability to form the complex was studied by radioimmunoprecipitation. It was found that N-terminal domains of gE and gI containing first 122 and 106 aa, respectively, were sufficient for the complex formation. PMID- 17131936 TI - Experimental infection of Hyalomma marginatum ticks with West Nile virus. AB - To define the possible role of Hyalomma marginatum ticks in the transmission of West Nile virus (WNV) in Portugal an experimental infection was established. Ticks were fed on viremic rabbits previously infected with WNV. In different developmental stage of H. marginatum virus isolation and detection of viral antigen and viral RNA were attempted. The oral infection rates were 3%, 33% and 75% for engorged larvae, nymphs and females after oviposition, respectively. Transstadial transmission rates for nymphs exposed to virus as larvae, for adults exposed as larvae, and for adults exposed as nymphs were 33%, 11% and 46%, respectively. No evidence of transovarial transmission was obtained. Ticks in the stages of nymphs and adults were able to transmit the infection to uninfected hosts. This study demonstrated that H. marginatum could be involved in the natural circulation of WNV in Portugal. PMID- 17131937 TI - Evaluation of clinical specimens for influenza A virus positivity using various diagnostic methods. AB - The diagnostic method for Influenza A virus, utilizing the SERION ELISA Antigen kit (SERION EIA), if results were evaluated according to the manufacturer's instructions, has repeatedly failed to detect a great number of clinical samples positive by virus isolation and RT-PCR. Therefore we compared the SERION EIA with the one-step 44/107L-Px immunocapture enzyme immunoassay (44/107L-Px EIA), developed in our laboratory (Tkacova and Vareckova, J. Virol. Methods 60, 65-71, 1996). Seventy-three clinical specimens, of which 65 were positive by virus isolation (used as reference method), were tested by both EIAs. By the SERION EIA, out of the 65 reference-positive samples only 8 (12%) were positive, 5 (8%) were ambiguous, and 52 (80%) were negative, which corresponded to the sensitivity of 12%. On the contrary, the sensitivity of the 44/107L-Px EIA was 74%. However, the calculation of cut-off values for the evaluation of positivity of clinical specimens in these two assays were not the same. If the evaluation procedure used for the 44/107L-Px EIA was applied to the SERION EIA, the sensitivity and the specificity of both EIAs became comparable, namely 71% and 100% for the SERION EIA and 74% and 100% for the 44/107L-Px EIA, respectively. From these results it follows that not the detection ability of the SERION EIA, but the evaluation procedure recommended by its manufacturer led to a loss of large number of positive specimens. PMID- 17131938 TI - pH modulating activity of ion channels of influenza A, B, and C viruses. AB - The BM2 and NB proteins of Influenza B virus (the B virus) and the CM2 protein of Influenza C virus (the C virus) are structural homologs of the M2 protein of Influenza A virus (the A virus). It was shown recently that CM2 in vitro forms a voltage-activated ion channel permeable to chloride ion (Hongo et al., Arch. Virol. 149, 35-50, 2004). To demonstrate a possible pH modulating activity of BM2, NB and CM2, the latters were co-expressed with a pH-sensitive hemagglutinin (HA) of the A virus. BM2 was able to replace functionally M2 and prevented the A virus HA from adopting its low-pH conformation during transport to the cell surface. In contrast, NB had a negative effect on the quality of the co-expressed HA and was unable to modulate the pH in the trans-Golgi network (TGN) and to protect HA. A pH modulating activity was also demonstrated for CM2, but it was much lower than that of M2. PMID- 17131939 TI - Cloning, expression and identification of the gene of human single-chain variable fragment antibody against Hepatitis B virus surface antigen. AB - Expression of single-chain variable fragment (scFv) antibodies on the surface of bacteriophage is widely used to prepare antibodies with pre-defined specificities. A phage antibody library containing the gene for scFv antibody against Hepatitis B virus surface antigen (HBsAg) was panned with HBsAg immobilized on microtiter plate wells. After five rounds of panning 30 phage clones specific to HBsAg were obtained and one selected clone was sequenced. It was found to consist of 789 bp and its amino acid sequence and specifically detected the respective antigen in the patients but not in healthy persons. PMID- 17131940 TI - Partial molecular characterization of a mild isolate of Grapevine fanleaf virus from South Moravia, Czech Republic. AB - An atypical mild isolate HV5 of Grapevine fanleaf virus (GFLV) was found in a South Moravian viticulture region in Czech Republic. Partial sequence of its RNA2 was determined and compared with available sequences of typical GFLV isolates. Two genomic regions, namely a 814 nt-long one spanning the movement protein (MP) gene and a 5'-part of the coat protein (CP) gene. and a 1426 nt-long one covering a part of the CP gene and the adjacent 3'-non-coding region (3'-NCR) were analyzed. Although no HV5-specific molecular features could be found in the two regions, marked differences were observed in the 3'-NCR. There was a 54 nt-long portion in which the sequence identity of some compared isolates was only 54.7%. Moreover, an unique one-nucleotide deletion occurred in the HV5 3'-NCR. These changes were also reflected in the predicted RNA secondary structure of this region. Particular biological behavior of GFLV HV5 isolate, namely a symptomless infection, could be related to the observed molecular differences. PMID- 17131941 TI - Molecular characterization of Czech and Chinese leek yellow stripe virus isolates from garlic. AB - Using an RT-PCR specific for nuclear inclusion b (NIb) and coat protein (CP) genes of Leek yellow stripe virus (LYSV) we detected two Czech (LYSV-5CZ and LYSV 22CZ) and one Chinese (LYSV-16) isolate of LYSV in garlic plants. The RT-PCR products were cloned and sequenced. Phylogenetic analysis based on deduced amino acid sequence of NIb-CP region placed the Czech isolates in the group I and the Chinese isolate in the group II of LYSV. PMID- 17131942 TI - The possible origin of recent human SARS coronavirus isolate from China. PMID- 17131943 TI - Vitamin D status and the metabolic syndrome. AB - The identification of vitamin D receptor expression in different tissues suggests a widespread role for vitamin D action beyond its classical function in bone and mineral metabolism. Recently, the importance of vitamin D status as a risk factor in the development of metabolic syndrome has been the focus of several studies. PMID- 17131944 TI - Building bones in babies: can and should we exceed the human milk-fed infant's rate of bone calcium accretion? AB - Increasing calcium absorption and bone calcium accretion to levels above those achieved by human milk-fed, full-term infants is possible with infant formulas. However, no data support such a goal or suggest that it is beneficial to short- or long-term bone health. Small differences in the bioavailability of calcium between infant formulas are unlikely to have long-term consequences. Long-term studies of the effects of infant feeding type on ultimate bone mass are needed. For now, the vitamin-replete breast-fed infant's rate of calcium accretion during the first year of life should be the standard targeted for infant formulas. PMID- 17131945 TI - Medical foods: products for the management of chronic diseases. AB - Medical foods are a specific category of therapeutic agents created under the Orphan Drug Act of 1988, which separated medical foods from drugs for regulatory purposes. Products in this category share the requirements that they are intended for the nutritional management of a specific disease, are used under the guidance of a physician, and contain ingredients that are generally recognized as safe (GRAS). An example of medical foods are formulations intended to manage patients with inborn errors in amino acid metabolism. Newer medical foods are designed to manage hyperhomocysteinemia, pancreatic exocrine insufficiency, inflammatory conditions, cancer cachexia, and other diseases. PMID- 17131946 TI - Effects of intended weight loss on morbidity and mortality: possible explanations of controversial results. AB - Long-term, population-based, observational studies have shown that intended weight loss does not always reduce the mortality associated with obesity. The effects of weight loss on mortality may be a balance between the effects of the loss of harmful abdominal and ectopic fat mass and the loss of beneficial peripheral subcutaneous fat mass and lean body mass. PMID- 17131947 TI - Catching up with our neighbors. PMID- 17131948 TI - The Upper Midwest Health Study: a case-control study of primary intracranial gliomas in farm and rural residents. AB - Since several studies indicated that farmers and agricultural workers had an excess risk of brain cancer, the National Institute for Occupational Safety and Health initiated the Upper Midwest Health Study to examine risk of intracranial glioma in the non-metropolitan population. This population-based, case-control study evaluated associations between gliomas and rural and farm exposures among adults (ages 18 to 80) in four upper midwestern states (Iowa, Michigan, Minnesota, Wisconsin). At diagnosis/selection, participants lived in non metropolitan counties where the largest population center had fewer than 250,000 residents. Cases were diagnosed 1 January 1995 through 31 January 1997. Over 90% of 873 eligible ascertained cases and over 70% of 1670 eligible controls consented to participate. Participants and nonparticipants, evaluated for "critical questions" on main and refusant questionnaires, differed significantly in farming and occupational experience, ethnicity, education, and lifestyle. The 1,175 controls were more likely than the 798 cases to have reported ever drinking alcohol (77% vs. 73%, adjusted odds ratio (OR) 0. 73, 95% confidence interval (CI) 0.59-0.92) and having had panoramic dental x-rays (34% vs. 29%, OR 0. 75, CI 0.61-0.92). Controls spent a greater percentage of their lives in non metropolitan counties (78% vs. 75%, OR 0.81, CI 0.67-1.09). Among ever-farmers, controls were more likely to have had exposure to farm insecticides (57% vs. 50%, OR 0.75, CI 0.59-0.95) and farm animals (96% vs. 91%, OR 0.48, CI 0.25-0.90). Moving to a farm as an adolescent (ages 11 to 20) vs. as an adult was associated with a greater risk of glioma. In our study sample, farm or rural residence and summary farm exposures were associated with decreased glioma risk. However, nonparticipation by never-farming eligible controls could have affected results. Comparisons of farm chemical exposures may clarify associations between farming and glioma that others have reported. PMID- 17131949 TI - National AgrAbility Project impact on farmers and ranchers with disabilities. AB - The impact of AgrAbility was evaluated through a survey of farmers and ranchers with disabilities who have been served by AgrAbility. The general demographics of the client population and assistance received were evaluated. Other information gathered included client ability pre- and post-onset of a disability and implications of self-reported outlook for the future. Eight states with AgrAbility programs participated in this cooperative survey with the National AgrAbility Project, with a 58.7% response rate (N = 618). The client population was mostly male (85.2%) with an average age of 53.3 with many working full-time (42.4%), part-time (27.6%), only off-farm (3%), or both off and on the farm (27%) in predominately row-crop (58.2%), cattle (not dairy) (46.6%), and hay or forage (41.4%) operations. Nearly half (48.2%) of the clients reported that the origin of the disability was due to a chronic health condition, as opposed to an injury. The majority of clients reported receiving information referring them to a funding source (42.0%) and receiving technical assistance with modifications around the farm or ranch (41.3%). Only two areas of farm operation were reported to have increased after the onset of disability (farm office from 43.8% to 61.2% and household chores from 30.9% to 36.0%). Field machinery operation continues to be the most common activity on the farm, with 73.3% reporting operating field machinery after the onset of disability. The present sample was more optimistic than expected. From a simultaneous multiple linear regression analysis, the factors contributing to positive future outlook include: ability to manage one's chores, machinery, and farm, F (5, 387) = 34.91, p < 0.001). Implications for safety professionals are included. PMID- 17131950 TI - Shock and vibration data acquisition system for off-road vehicle operator health and safety assessment. AB - A data acquisition software and hardware system was developed for acquiring geo referenced shock and vibration data using National Instruments' LabView graphical programming language. This was used in conjunction with a modular data acquisition and signal conditioning system and a Differential Global Positioning System (DGPS) receiver. A prototype vehicle obstacle course, which introduced spatially varying shock events to vehicles as they traversed the course, was constructed. Obstacles consisted of both repetitious and single discrete events. A series of investigations was conducted on the obstacle course to evaluate the performance and characteristics of the DAQ system and the tractor when exposed to shock and vibration events. Spectral and time domain plots obtained from the geo referenced data acquisition system (GDAQ) system under static, highway, and off road obstacle course conditions were evaluated to demonstrate that the system performed as expected. The migration of experiments from laboratory to field gave confidence that this system could be used to collect shock and vibration data over a wide range of frequencies. The use of geo-referenced data records proved beneficial in isolating and extracting data segments of interest from a continuous data record. PMID- 17131951 TI - Nonfatal injuries to household youth on racial minority-operated farms in the U.S., 2000. AB - The National Institute for Occupational Safety and Health (NIOSH) estimated that 32,808 nonfatal injuries occurred to youth less than 20 years of age on U.S. farms during 1998. These data, however, do not allow for the identification of minority farm operators. The Minority Farm Operator Childhood Agricultural Injury Survey (M-CAIS) was conducted to provide an overview of the number of youth on minority-operated farms and their associated farm-related injuries during 2000. M CAIS was conducted by the USDA for NIOSH through a telephone survey of 49,270 minority-operated farms identified in the 1997 Census of Agriculture. These minority-operated farms included four racial categories (black, Asian, Native American, and other) and operators of Hispanic ethnicity. This study included only racial minority-operated farms for analysis, white Hispanic farms were excluded. In 2000, there were an estimated 28,577 youth living on U.S. farms operated by racial minorities. In that year, these youth sustained an estimated 348 nonfatal injuries. Males accounted for 245 (70%) of the injuries to household youth. The majority of all injuries to household youth (247, 71%) occurred on livestock operations. Native American household youth accounted for both the largest number of injuries (177) and the highest rate of injury (24.0/1,000 household youth) on these farms. M-CAIS data indicated significant variation in injury rates among specific racial categories. Results of the M-CAIS suggest the need for prevention strategies to address issues found within these specific sub populations of the agricultural community. PMID- 17131952 TI - A survey of tractors and rollover protective structures in Washington State. AB - A survey of farms in Washington State was conducted to determine tractor characteristics and the presence of rollover protective structures (ROPS) in a state with more inclusive rules on tractor retrofitting than federal regulations. A total of 544 valid surveys were completed from a proportional random sample across different types of farms. Responders indicated that 58% of tractors overall were equipped with ROPS, and 42% of the tractors without ROPS were exempt from the state rules. Seatbelts on tractors equipped with ROPS were reportedly used "sometimes" or more 30% of the time, and 17% of these tractors had no seatbelt installed. Tractors used for row crop farming were significantly more likely to be equipped with ROPS than those used for tree, vine, or hops farming. Older tractors were used for fewer hours, were less likely to be ROPS-equipped, and were less likely to be operated while wearing a seatbelt. The results were consistent with a positive effect of the Washington State ROPS requirements, demonstrated by the increased percentage of ROPS-equipped pre-1976 tractors, as compared to other states, and by the difference between ROPS-equipped tractors in exempt and non-exempt types of farming. The results point to the need for prevention activities to increase seatbelt use on ROPS-equipped tractors, and for further development of practical protection for tractors operating under overhead obstacles. PMID- 17131953 TI - The community benefits of farm safety day camps. AB - In addition to the direct impact of a farm safety day camp on its participants' knowledge and safety awareness, there are extended and indirect effects that occur through the wider dispersion of information and the involvement and cooperation of community members. Reports completed by 228 coordinators of farm safety day camps, report forms completed by 5,037 volunteers at farm safety day camps, and telephone interviews with 924 parents of farm safety day camp participants were analyzed for evidence of the impact of the camps beyond the immediate knowledge gained by the children who participated. These data indicate that the indirect benefits to a community include enhanced safety awareness within the wider community as children and adult volunteers disseminate the information they learned, as well as enhanced community strength and cohesiveness resulting from the cooperation of many individuals and organizations in achieving a common goal. PMID- 17131954 TI - The ethics of medicine and nursing. PMID- 17131955 TI - Overview of hepatitis C and skin. AB - Hepatitis C (HCV) is the most common cause of chronic liver disease and hepatocellular carcinoma, as well as the leading indication for liver transplantation in the Western world. For many patients, cutaneous manifestations may be the only, the earliest, or the most apparent sign of the underlying infection. The dermatologic manifestations of HCV infection are reviewed. PMID- 17131956 TI - Allergic contact dermatitis: early recognition and diagnosis of important allergens. AB - Allergic contact dermatitis (ACD) is an important disease with high quality of life and economic impact. Patch testing is the procedure by which identification of the cause of ACD can be elicited. Proper performance of the test, from taking an appropriate patient history to placing the correct allergens to evaluating and educating the patient, is of utmost importance. The purpose of this article is to highlight common allergens encountered in our environment, to increase awareness for this important disease, and to underscore the importance of this testing modality. An early index of suspicion can lead to appropriate testing, diagnosis, avoidance, and cure. PMID- 17131957 TI - What's your assessment? Pernio. PMID- 17131959 TI - Cutaneous cryosurgery. AB - Cryosurgery is a therapeutic modality useful for treating cutaneous benign, premalignant, and malignant lesions. It consists of lowering lesional temperature below a critical level to cause its destruction. Lowering skin temperatures without the intent to directly cause lesional destruction (to trigger an immune mechanism, for example) is termed cryotherapy. PMID- 17131960 TI - Case study on linear focal elastosis. AB - Linear focal elastosis is an uncommon dermal elastic disorder, characterized by palpable, linear bands clinically and an increase in abnormal elastic tissue histologically. Two similar cases are presented and discussed. PMID- 17131961 TI - Where did all of the windows go? PMID- 17131962 TI - The right staff: a primer of practice for medical assistants and aestheticians. PMID- 17131963 TI - Cutaneous manifestations of systemic disease: leukemia/lymphoma, systemic malignancy, hypothyroidism, diabetes mellitus, and neurofibromatosis. PMID- 17131964 TI - 'It hurts when I walk:' venous stasis disease--etiology and assessment. PMID- 17131965 TI - Pyoderma gangrenosum. PMID- 17131966 TI - Erythema multiforme. PMID- 17131967 TI - Impact of a mass immunization campaign to control an outbreak of severe respiratory infections in Nunavik, northern Canada. AB - OBJECTIVES: In spring 2002, a mass immunization campaign using a 23-valent pneumococcal polysaccharide vaccine (23-PPV) was launched in order to control an outbreak of severe pneumonia caused by a virulent clone of serotype 1 Streptococcus pneumoniae in Nunavik, Quebec, Canada. The objective of the study was to evaluate the impact of this campaign on the incidence of invasive pneumococcal disease (IPD) and hospitalizations possibly associated with pneumococcal infections (HPAPI) in the mostly Inuit population aged 10 to 64 years. STUDY DESIGN: Retrospective analysis of surveillance and administrative data. METHODS: Multivariate Poisson model comparing the frequency rates of selected outcomes before the outbreak, during the outbreak, and after implementation of the mass immunization program. RESULTS: The reported incidence of serotype 1 IPD decreased markedly after the implementation of the vaccination campaign (rate ratio = 0.16; p < 0.002). The frequency of HPAPI and the mean duration of hospital stay also decreased. However, vaccine failures were documented and the HPAPI rate remained higher than in the period prior to the outbreak. CONCLUSIONS: Although 23-PPV contributed to control the outbreak, better vaccines are needed for the prevention of infections caused by serotype 1 S. pneumoniae. PMID- 17131968 TI - Bullying among Greenlandic schoolchildren: development since 1994 and relations to health and health behaviour. AB - OBJECTIVES: The objective was to examine the development in the prevalence of bullying behaviours among Greenlandic schoolchildren and the association with health outcome and health behaviour. STUDY DESIGN: The study was based on three school surveys among Greenlandic schoolchildren contributing to the Health Behaviour in School-aged Children (HBSC) survey, a WHO collaborative study. The surveys were carried out in Greenland in 1994, 1998 and 2002, with respective participations of 1322, 1648 and 891 pupils of 11, 13 and 15 years of age. METHODS: The trends in bullying behaviours from 1994 to 2002 was analysed by means of descriptive statistics. The strengths of associations in the patterning of the bullying behaviours in their relation to health indicators (physical symptoms, psychological well-being and smoking and alcohol use) were analysed by means of logistic regression. RESULTS: There has been an increase in the occurrence of bullying among Greenlandic schoolchildren since 1994, and significant changes have occurred in the different types of bullying behaviours. Consistent patterns were observed between types of bullying behaviours, and health behaviour, since pupils engaged in bullying were more likely to be smokers and to have been drunk several times. Strong associations were observed between disliking school and being engaged in bullying, whether this was as a victim, a bully, or both. There was no clear patterning of associations when it came to health indicators, except for significantly higher odds of stomach ache for the bullies, and sleeping difficulties and low self-rated health for pupils both being bullied and bullying others. CONCLUSIONS: Being engaged in bullying is widespread among Greenlandic schoolchildren and is found to be associated with disliking school and detrimental health behaviours. PMID- 17131969 TI - HIV/AIDS: testing and risk behaviors among British Columbia's rural Aboriginal population. AB - OBJECTIVES: Over the last decade, there has been growing concern in the public health sector over the spread of Human Immunodeficiency Virus (HIV) in Canada's Aboriginal population. However, there continues to be a general lack of HIV awareness and its risk factors in Aboriginal communities. STUDY DESIGN AND METHODS: This study investigated HIV testing patterns, perceptions and risk factors within 7 community organizations through the use of face-to-face interviews. The objectives of this research project were to 1) describe the prevalence of HIV testing; 2) describe issues concerning confidentiality related to the HIV testing; 3) identify the prevalence of risk factors for HIV transmission; and, 4) build research capacity in the Aboriginal community. RESULTS: Two hundred and nineteen Aboriginal persons participated in interviews. Off-reserve residents (pOR: 0.48, 95% CI: 0.29 - 0.80) were significantly more likely to use illicit drugs than reserve residents. Sixty percent of participants had reported undergoing HIV testing at some point. Two-spirited participants (pOR: 16.1, 95% CI: 2.13-121.06), those who previously tested for a Sexually Transmitted Disease (pOR: 2.94, 95% CI: 1.73-4.98), those currently using cocaine (pOR: 3.88, 95% CI: 1.25 - 12.0), and those who reported to "never", or "some of the time" use clean needles (pOR: 11.0, 95% CI: 1.36- 88.66) were significantly more likely to undergoing HIV testing. On-reserve residents (pOR: 0.55, 95% CI: 0.35-0.87) and respondents older than 40 years of age (pOR: 0.40, 95% CI: 0.19 - 0.86) were less likely to undergo HIV testing. CONCLUSIONS: Participants with HIV risk factors are more likely to be tested for HIV in rural areas and confidentiality issues were not a barrier to testing for most participants. Off reserve residents were more likely to undergo HIV testing, the reasons for which require additional research. Finally, public health units are often under utilized as locations to seek testing. PMID- 17131970 TI - Obesity, adiposity, physical fitness and activity levels in Cree children. AB - OBJECTIVES: To describe the levels of obesity, adiposity measures, physical activity and fitness in Cree children aged 9-12 years. STUDY DESIGN: Cross sectional survey. METHODS: The study took place in northern Quebec, Canada. Height, body mass, waist circumference and five skinfold thicknesses were measured. Physical activity was assessed by having children wear a pedometer for two days. Children performed the 20-metre shuttle run test (SRT) to determine their physical fitness level. RESULTS: Of 82 participating children, 33% were overweight (but not obese) and 38% were obese according to an international reference. The mean sum of five skinfold measures exceeded the 95th percentile of Canadian children. Compared with the Third National Health and Nutrition Examination Survey, the majority of children exceeded the 85th percentile for waist circumference (62%) and at the suprailiac (80%), subscapular (72%), and triceps (54%) skinfold sites. 90% of children scored below the 20th percentile in the SRT compared with normative data from Quebec children. Based on pedometer scores, only 49% of children were sufficiently active. CONCLUSIONS: There is a high prevalence of overweight and central adiposity in this population, with low physical activity and fitness levels. This profile may result in adverse health outcomes. PMID- 17131971 TI - Traditional and market food access in Arctic Canada is affected by economic factors. AB - OBJECTIVES: This study aimed to evaluate the access that Indigenous women have to traditional and market foods in 44 communities across Arctic Canada. STUDY DESIGN: This secondary data analysis used a cross-sectional survey of 1771 Yukon First Nations, Dene/Metis and Inuit women stratified by age. METHODS: Socio cultural questionnaires were used to investigate food access and chi-square testing was used to ascertain the distribution of subject responses by age and region. RESULTS: There was considerable regional variation in the ability to afford adequate food, with between 40% and 70% saying they could afford enough food. Similarly, regional variation was reflected in the percentage of the population who could afford, or had access to, hunting or fishing equipment. Up to 50% of the responses indicated inadequate access to fishing and hunting equipment, and up to 46% of participants said they could not afford to go hunting or fishing. CONCLUSIONS: Affordability of market food and accessibility to hunting and fishing in Arctic Canada were major barriers to Indigenous women's food security. PMID- 17131972 TI - Unchanged asthma prevalence during 1990-1999 in rural Alaska Natives. AB - OBJECTIVES: The prevalence of asthma has been increasing throughout the world, but the reasons for the increase are unclear. Some have hypothesized that the increase is due to industrial and agricultural pollutants in urban and rural areas, respectively. The objective of this research was to determine if the prevalence of asthma has increased in a remote area of Alaska where the population lives a subsistence lifestyle and is not exposed to such pollution. STUDY DESIGN: Retrospective review of medical records to determine the prevalence of asthma. METHODS: We reviewed medical records of 1200 children, aged 0-10 years, who lived in the Yukon-Kuskokwum Delta (YKD) region of western Alaska between 1990 and 1999. The entire YKD population receives health care from a single medical system, so records provide a complete picture of each patient's health care. Data collected from the medical records included demographics, and the presence or absence of a diagnosis of asthma or reactive airway disease (RAD). RESULTS: Over the 10-year period from 1990-1999, there was no significant change in the percentage of children who had a diagnosis of asthma (2.0% in 1990 and 3% in 1999), or RAD (9.6% in 1990 and 9.6% in 1999). CONCLUSIONS: In a population of children not exposed to urban industrial, or rural agricultural pollutants, there was no change in the prevalence of wheezing or asthma between 1990 and 1999. PMID- 17131973 TI - The role played by a former federal government residential school in a First Nation community's alcohol abuse and impaired driving: results of a talking circle. AB - OBJECTIVES: The study's objective was to better understand alcohol abuse and impaired driving behaviors in a First Nations community as it reflects systemic issues linked to historical, family and community experiences. STUDY DESIGN AND METHODS: Fifteen 18- to 29-year-old drivers participated in an exploratory eight hour Talking Circle held according to traditional cultural practice. Four First Nations researchers, trained in Talking Circle protocol, and a Band Elder facilitated the data collection, data analysis according to emerging themes, and data verification. RESULTS: Federal government residential schools contribute to intergenerational effects which impact impaired driving in a northern First Nations community. Traditional parental role modeling has changed dramatically. Rather than guide children through a communally shared development process, many parents now expect their children to assume adult roles by expecting them to take care of their guardians when they drink excessive amounts of alcohol. Because a wall of silence exists between the young and old, many young people seek refuge with friends and peers, who subsequently influence them to abuse alcohol and engage in impaired driving. Many older Band members no longer serve as leaders for young people. Instead, they behave like peers and engage in activities that facilitate alcohol abuse and impaired driving. CONCLUSIONS: Historical institutions like federal government residential schools have contributed to systemic socio cultural problems which influence alcohol abuse and impaired driving. Hence there is a need for community-based intervention strategies that promote cultural healing. The healing journey can start with First Nations communities providing their people opportunities to share their stresses and traumas in supporting and nurturing environments. PMID- 17131974 TI - Changes in reindeer herding work and their effect on occupational accidents. AB - OBJECTIVES: The aims of the study were to investigate the risk of occupational accidents in reindeer herding work in the long run, and to find out if the changes of work processes caused by joining the European Union in 1995 can be seen in accident statistics. STUDY DESIGN AND METHODS: The information on reindeer herders' occupational accidents was obtained from the Farmers' Social Insurance Institution. The development of accidents, reindeer herders and handled animals was studied in 1991-2004. The work phase and contact-mode of the compensated accidents during the periods of 1991-1994 (N = 514) and 2001-2004 (N = 411) were compared. RESULTS: About 100 accidents of reindeer herders are compensated by the Farmers' Social Insurance Institution annually. The number of accidents much better with the number of reindeer handled than with that of insured reindeer herders. The changes in work processes could be seen as trends in the classification of accidents, but the differences were not statistically significant. In feeding tasks, both on the terrain and on farms, accidents had slightly increased, whereas in slaughtering they had decreased. Reindeer-caused accidents are now a bit more frequent than earlier. CONCLUSIONS: Attention should now be paid to the use of personal protective equipment when driving all-terrain vehicles and to safe working habits when in close contact with reindeer. PMID- 17131975 TI - Environmental medicine in an arctic perspective: "The Arctic Dilemma" revisited. PMID- 17131976 TI - Notes from Nepal: reflections of a medical student on shamans, lamas, serpents, and fortunes. PMID- 17131977 TI - Holistic medicine in Nepal. PMID- 17131978 TI - Practiced by millions, prayer is worthy of more study. PMID- 17131979 TI - A home study-based spirituality education program decreases emotional distress and increases quality of life--a randomized, controlled trial. AB - CONTEXT: Although epidemiological studies have reported protective effects of religion and spirituality on mental health, it is unknown whether spirituality can be used as an intervention to improve psychological well-being. OBJECTIVE: To evaluate the efficacy of a home study-based spirituality program on mood disturbance in emotionally distressed patients. DESIGN, SETTING, AND PARTICIPANTS: A non-blinded, randomized, wait list-controlled trial of 165 individuals with mood disturbance [score of >40 on the Profile of Mood States (POMS)] were recruited from primary care clinics in a Canadian city between August 2000 and March 2001. INTERVENTIONS: Participants were randomized to a spirituality group (an 8-week audiotaped spirituality home-study program), a mindfulness meditation-based stress reduction group (attendance at facilitated classes for 8 weeks), or a wait-list control group (no intervention for 12 weeks). MAIN OUTCOME MEASURES: Primary outcomes were mood disturbance, measured using POMS, and quality of life, measured using the SF-36, a short-form health survey with 36 questions. The POMS and the SF-36 were completed at baseline, at 8 weeks, and at 12 weeks. RESULTS: At the end of the 8-week intervention period, the mean POMS score improvement was -43.1 (-45.7%) for the spirituality group, 22.6 (-26.3%) for the meditation group, and -10.3 (11.3%) for the control group (P<.001 for spirituality vs control group; P=.034 for spirituality vs meditation group). Mean improvement in the SF-36 mental component summary score was 14.4 (48.6%) for the spirituality group, 7.1 (22.3%) for the meditation group, and 4.7 (16.1%) for the control group (P<.001 for spirituality vs control group; P=.029 for spirituality vs meditation group). At 12 weeks, POMS and SF-36 scores remained significantly different from baseline for the spirituality group. PMID- 17131980 TI - The efficacy of distant healing for human immunodeficiency virus--results of a randomized trial. AB - BACKGROUND: While data are conflicting, studies have appeared in the literature suggesting that mental intentions sent from a distance (eg, intercessory prayer, spiritual healing) can possibly influence clinical outcomes in patients suffering from an array of medical conditions. The purpose of this study was to examine the potential efficacy of distant healing in a population of patients with human immunodeficiency virus (HIV)/aquired immune deficiency syndrome (AIDS). METHODS: One hundred fifty-six patients with a history of AIDS category C and at least one AIDS-defining opportunistic infection were randomized to 1 of 3 study arms: (1) 10 weeks of prayer/distant healing from professional healers, (2) 10 weeks of prayer/distant healing from nurses with no prior training or experience in distant healing, or, (3) no distant healing. RESULTS: No significant treatment effects of distant healing were observed for either professional healers or nurses on any of the primary or secondary outcomes. Despite being blind to group assignment, subjects receiving distant healing (from healers or nurses) were significantly more likely to guess that they had been receiving healing than were subjects randomized to the no-treatment control group. CONCLUSIONS: Distant healing or prayer from a distance does not appear to improve selected clinical outcomes in HIV patients who are on a combination antiretroviral therapy. PMID- 17131981 TI - The effect of intercessory prayer on wound healing in nonhuman primates. AB - OBJECTIVES: This study was performed to examine the effects of intercessory prayer (IP) on wound healing and related physiological and behavioral factors in nonhuman primates. DESIGN: Twenty-two bush babies (Otolemur garnettii) with chronic self-injurious behavior (SIB) were stratified by wound severity and matched by total wound area. The animals were then randomized to IP and L tryptophan or L-tryptophan only for treatment of SIB and related wounds. The IP intervention was conducted in a double-blind, randomized manner. Prayer was conducted daily for 4 weeks. Initiation of prayer was coincident with the first day of L-tryptophan administration. Physiological and behavioral variables were assessed at baseline and end of study. RESULTS: Following IP/L-tryptophan treatment, prayer-group animals had a reduction in wound size compared to non prayer animals (P=.028). Prayer-group animals had a greater increase in red blood cells (P=.006), hemoglobin (P=.01), and hematocrit (P=.018); a greater reduction in both mean corpuscular hemoglobin (P=.023) and corpuscular volume (P=.008); and a reduction in wound grooming (P=.01) and total grooming behaviors (P=.04) than non-prayer-group animals. CONCLUSIONS: The results of this study are consistent with prior human trials of IP effectiveness, but suggest IP-induced health improvements may be independent of confounds associated with human participants. Findings may provide direction for study of the mechanisms of IP-induced health improvements in both human and animal models. PMID- 17131983 TI - Consciousness and nonlocality. PMID- 17131984 TI - Control of antigen presentation with a photoreleasable agonist peptide. AB - The immunological synapse is a specialized intercellular junction between a T cell and a target cell that orchestrates the engagement of receptors and ligands in space and time as a means of regulating function. Here we introduce a reagent for controlling the spatial and temporal presentation of natural antigen to T cells. Moth cytochrome c (88-103) peptide (MCC), an agonist to the murine T cell receptor AND when presented in the context of H2 IEk major histocompatibility complex (IEk), was synthesized with the side-chain amine of Lys99 conjugated to a photosensitive protecting group, 6-nitroveratryloxycarbonyl (NVOC). Cells plated on supported bilayers displaying mobile intercellular adhesion molecule-1 (ICAM 1) and NVOC-MCC loaded IEk did not form immunological synapses and exhibited low intracellular calcium levels, similar to cells presented with self-peptide. Irradiation with UV light was sufficient to restore agonist activity in situ. PMID- 17131985 TI - Activity-based substrate profiling for Gcn5-related N-acetyltransferases: the use of chloroacetyl-coenzyme A to identify protein substrates. AB - The Gcn5-related N-acetyltransferases (GNAT) comprise one of the largest enzyme superfamilies, with over 10 000 known members represented in all kingdoms of life. ChloroacetylCoenzymeA was prepared and demonstrated to be a substrate for several GNAT members. ChloroacetylCoA (ClAcCoA) is used by the Hat1 histone acetyltransferase to correctly acetylate histone H4 in a mixture of histone proteins. Chloroacetylation can be assessed by the subsequent reaction of the chloroacetylated product with thiol-containing compounds, including those with fluorescent or affinity (His8) tags. The bacterial RimL N-acetyltransferase also uses ClAcCoA to chloroacetyl the alpha-amino group of its cognate substrate, the ribosomal L12 protein, and this reaction can be observed in crude extracts. ChloroacetylCoA is a reagent that can be used to identify the unknown substrate(s) for this large family of functionally uncharacterized enzymes. PMID- 17131986 TI - Template-directed one-step synthesis of cyclic trimers by ADMET. AB - A trifurcated template, containing three secondary dialkylammonium ion recognition sites for encirclement by a dibenzo [24]crown-8-containing acyclic diene, is used to promote acyclic diene metatheses (ADMET) catalyzed by ruthenium alkylidene complexes, affording a cyclic trimer in 55% yield. Following this one step, threefold ADMET reaction, the resulting cyclic trimer was isolated by preparative HPLC and characterized by NMR spectroscopy and mass spectrometry. PMID- 17131987 TI - Critical guest concentration and complete tuning pattern appearing in the binary clathrate hydrates. AB - The concept of tuning phenomenon in binary hydrate systems has been suggested to enhance the gas storage capacity through molecular interactions. In this report, the existence of critical guest concentration (CGC) is investigated by means of spectroscopic methods. The existence of the critical guest concentration can act as a limiting factor in the application areas of binary hydrates. Therefore, it should be taken into account before applying this concept to application fields. In addition, further research on this concept using other hydrate systems is required to clarify the present findings. PMID- 17131988 TI - Hybrid gold/silica/nanocrystal-quantum-dot superstructures: synthesis and analysis of semiconductor-metal interactions. AB - We report on the synthesis and spectroscopic characterization of well-defined hybrid structures that consist of a gold core overcoated with a silica shell, followed by a dense monolayer of CdSe nanocrystal quantum dots (QDs). The dielectric silica spacer of a controlled thickness provides a simple means for tuning interactions between the QD emitters and the metal core. To illustrate this tunability, we demonstrate switching between QD emission quenching and enhancement by varying the silica shell thickness. Synthetic procedures developed here employ a final step of self-assembly of QDs onto the silica shell performed via simple titration of the QD solution with prefabricated core/shell Au/SiO2 particles. This approach allows us to perform an accurate quantitative analysis of the effect of the metal on the QD emission intensity. One important result of this analysis is that nonuniformity of nonradiative rates across the QD ensemble has a significant effect on both the magnitude and the shell-thickness dependence of the emission enhancement/quenching factors. PMID- 17131990 TI - A synthetic mimic of protein inner space: buried polar interactions in a deep water-soluble host. AB - A deep water-soluble cavitand was functionalized with a carboxylic acid directed toward the hydrophobic interior of the host. The buried salt-bridge interaction formed with a quinuclidium cationic guest was determined to be worth -3 kcal/mol using a free energy cycle. The strength of the interaction correlates well with buried salt bridges in proteins, indicating that the cavitand interior mimics the hydrophobic inner space of proteins. PMID- 17131989 TI - Unexpected acetylcholinesterase activity of cocaine esterases. AB - A series of proteins with cocaine esterase ability have been shown to hydrolyze acetylcholine with similar rate enhancements. Docking studies revealed that acetylcholine binds in a similar orientation as cocaine. These results suggest that acetylcholinesterase activity may be inherent to cocaine esterases. PMID- 17131991 TI - Azotolane liquid-crystalline polymers: huge change in birefringence by photoinduced alignment change. AB - Novel liquid-crystalline polymers (LCPs) containing a long azotolane moiety were synthesized, and the relationship between the chemical structures and the photoresponsive behavior was investigated. All homogeneously aligned LCP films exhibited extremely high values of birefringence (Deltan) in the wide wavelength range. Specifically, the azotolane LCP with two azobenzene units showed the most efficient change in the alignment of the azotolane moiety with a huge change in Deltan (>/=0.65). This means that the high value of Deltan in the homogeneously aligned state can be converted to the change in Deltan effectively. PMID- 17131992 TI - Self-assembly of coordination polymers: evidence for dynamic exchange between oligomers in solution and the isolation of a homochiral decagold(I) oligomer. AB - Reactions of the precursor molecules [Au2(mu-BINAP)(O2CCF3)2], 1a, racemic BINAP, 1b, S-BINAP (BINAP = 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl) with the easily exchanged linear bis(pyridine) ligand 1,2-trans-bis(4-pyridyl)ethylene (bipyen) gave the polymeric complex [{Au2(mu-R-BINAP)0.5(mu-S-BINAP)0.5(mu bipyen)}n](CF3CO2)2n, 2a, but either the polymer [{Au2(mu-S-BINAP)(mu bipyen)}n](CF3CO2)2n, 2b, or the remarkable oligomeric [Au10(mu-S-BINAP)5(mu bipyen)4(kappa1-bipyen)2](CF3CO2)10, 3, respectively. The type of oligomer 3 is a missing link in the ring-opening polymerization of macrocyclic coordination compounds. PMID- 17131993 TI - FEP-guided selection of bicyclic heterocycles in lead optimization for non nucleoside inhibitors of HIV-1 reverse transcriptase. AB - Monte Carlo simulations using free energy perturbation theory have been used to guide the selection of bicyclic heterocycles in the lead optimization of non nucleoside inhibitors of HIV-1 reverse transcriptase (NNRTIs). Good correlation is found between predicted and observed activities. Six compounds are reported with EC50 values below 20 nM for protection of human MT-2 cells against the cytopathogenicity of HIV-1. Striking variation in activity is found and analyzed for an isomeric pyrrolopyrimidine and pyrrolopyrazine pair. PMID- 17131994 TI - Homo- and hetero-[3]rotaxanes with two pi-systems clasped in a single macrocycle. AB - Here we present the first synthesis of a [3]rotaxane with two dumbbell components threaded through a single gamma-cyclodextrin macrocycle. This synthesis is carried out in two steps: first one dumbbell is synthesized threaded through the macrocycle to give a [2]rotaxane, then a second dumbbell is synthesized through the remaining cavity of the [2]rotaxane. We have synthesized a hetero- [3]rotaxane with one stilbene and one cyanine dye threaded through gamma cyclodextrin, which exhibits quantitative energy transfer between the two encapsulated dyes. The stilbene [2]rotaxane intermediate in this synthesis has a remarkably high affinity for suitably shaped hydrophobic guests in aqueous solution, facilitating the synthesis of [3]rotaxanes and suggesting possible applications in sensors. PMID- 17131995 TI - Probing biocatalytic transformations with CdSe-ZnS QDs. AB - CdSe/ZnS QDs enable the optical probing of the biocatalytic oxidation of tyrosine derivatives and of the scission of peptides by thrombin. CdSe/ZnS QDs were modified with tyrosine methyl ester or with a tyrosine-containing peptide. The tyrosine units were reacted with tyrosinase/O2 to yield the respective l-DOPA and quinone derivatives. The luminescence of QDs modified by the enzyme-generated quinone units is quenched. The quinone-functionalized peptide associated with the QDs was cleaved by thrombin, a process that restored the luminescence of the QDs. PMID- 17131996 TI - Asymmetric modular synthesis of highly functionalized medium-sized carbocycles and lactones via ring-closing metathesis of sulfoximine-substituted trienes. AB - A modular asymmetric synthesis of medium-sized carbocycles and lactones has been developed that affords highly substituted 7-, 9-, and 11-membered rings. The key steps are (1) the highly diastereoselective synthesis of sulfoximine-substituted homoallylic alcohols from allylic sulfoximines and unsaturated as well as saturated aldehydes, (2) an E-stereoselective alkylation and hydroxyalkylation of sulfoximine-substituted alkenyllithium derivatives, (3) the esterification of sulfoximine-substituted homoallylic alcohols, and (4) the ring-closing metathesis reaction of sulfoximine-substituted trienes with the ruthenium catalyst 8. Two examples for the further synthetic elaboration of the sulfoximine-substituted carbocycles are provided. The selective cleavage of the tert-butyldimethylsilyl group of 12 in the presence of the triethylsilyl group afforded the allylic alcohol 18 which was oxidized to enone 19. A cross-coupling reaction of the sulfoximine-substituted carbocycle 9 with LiCuMe2 furnished the methyl substituted derivative 20. PMID- 17131997 TI - Ethidium and proflavine binding to a 2',5'-linked RNA duplex. AB - Despite over 40 years of physical investigations, fundamental questions persist regarding the energetics of RNA and DNA intercalation. The dramatic unwinding of a nucleic acid duplex upon intercalation immediately suggests that the nucleic acid backbone should play a significant role in dictating the free energy of intercalation. However, the contribution of the backbone to intercalation free energy is difficult to appreciate given the intertwined energetics associated with intercalation (e.g., pi-pi stacking and solvent effects). Fluorescence titrations were used to determine the association constants of two known intercalators, proflavine and ethidium, for duplex 2',5'-linked RNA. Proflavine was found to bind 2',5' RNA with an association constant 25-fold greater than that measured for standard, 3',5'-linked RNA. In contrast, ethidium binds 2',5' RNA less favorably than standard RNA. PMID- 17131998 TI - Stereoselective Lewis acid-catalyzed alpha-acylvinyl additions. AB - Silyloxyallenes derived from alpha-hydroxypropargylsilanes undergo efficient addition to aldehydes with catalytic amounts of Lewis acids. The allenes are accessed from the corresponding propargylsilanes in a base-catalyzed 1,2-Brook rearrangement/SE2' process. Enantioenriched propargylsilanes are synthesized by a new zinc-promoted addition of alkynes to acylsilanes in up to 74% ee. This work demonstrates that conversion to the silyloxyallenes occurs with minimal erosion in optical activity. The use of a chiral chromium(III) Lewis acid effects the catalytic asymmetric addition of racemic silyloxyallenes to aromatic aldehydes in up to 92% ee. The overall reaction is broad in scope and accommodates a wide variety of aromatic and aliphatic substituents on both the propargylsilane and aldehyde. PMID- 17131999 TI - Extraordinary atomic mobility of Au{111} at 80 Kelvin: effect of styrene adsorption. AB - We describe how the presence of styrene, a weakly adsorbed molecule, dramatically restructures the Au{111} surface at temperatures as low as 80 K. The restructuring manifests itself both in mobility of step-edge atoms, as well as changes in the position of the herringbone reconstruction over time. These effects are explained in terms of the preferential adsorption sites of styrene allowing it to assist in atom detachment from step edges, as well as lowering of the energetic barrier for movement of the herringbone reconstruction. This work has important consequences for studies in which Au is used as a support for or as an electrical contact to molecules. PMID- 17132000 TI - Enhanced oxide ion conductivity in stabilized delta-Bi2O3. AB - The substitution of Re into Bi2O3 allows stabilization of the delta-Bi2O3 structure by additional substitution of any lanthanide ion to give, for example, phases of composition Bi12.5La1.5ReO24.5. Some of these phases have been found to show exceptionally high oxide ion conductivity at low temperatures, ca 10-3 S cm 1 at 300 degrees C. The phases show a significant structural difference from other delta-Bi2O3 phases previously reported, with interstitial anion sites displaced further from the ideal fluorite position, (1/4,1/4,1/4). PMID- 17132001 TI - Unambiguous determination of the ionization state of a glycoside hydrolase active site lysine by 1H-15N heteronuclear correlation spectroscopy. AB - We have investigated the lysine side chain amines in the 34 kDa catalytic domain from Cellulomonas fimi beta-(1,4)-glycosidase Cex (or CfXyn10A) using 1H-detected 15N heteronuclear correlation NMR spectroscopy. Signals from the 1Hzeta ( approximately 8 ppm) and 15Nzeta ( approximately 35 ppm) of Lys302 in the unmodified enzyme and Lys47 in a trapped cellobiosyl-enzyme intermediate were detected in a 1H-15N HMQC spectrum (pH 6.5 and 30 degrees C). The amine of Lys302 forms a buried ion pair, and that of Lys47 is hydrogen bonded to the cellobioside. Both lysines are positively charged, as unambiguously demonstrated by the splitting of their 15Nzeta signals into quartets (|1JNH| approximately 75 Hz) in a 1H-15N HSQC spectrum recorded without 1H decoupling during 15N evolution. Qualitative insights into the dynamic properties of these lysines are also provided by the deviations of their quartet intensity ratios from that of approximately 3:1:1:3 expected for a highly mobile amine. On the basis of the observed ratios of approximately 1:1:1:1 for the quartet of Lys302 and approximately 0.5:1:1:0.5 for Lys47, the amine of the latter active site residue is most rigidly positioned. Signals from at least 8 and 10 additional positively charged, mobile amines in Cex were observed at 10 degrees C and pH 6.5 and 5.6, respectively. By using conditions of reduced temperature, slightly acidic pH, and low general base concentrations, as well as water flipback pulses to minimize the effects of hydrogen exchange, 1H-15N correlation experiments provide a sensitive route to directly investigate the charge states and dynamic properties of the N terminal and side chain amines in proteins and protein complexes. PMID- 17132002 TI - Metal-ligand cooperation in C-H and H2 activation by an electron-rich PNP Ir(I) system: facile ligand dearomatization-aromatization as key steps. AB - Unusual reactions are reported, in which the aromatic PNP ligand (PNP = 2,6-bis (di-tert-butylphosphinomethyl)pyridine) acts in concert with the metal in the activation of H2 and benzene, via facile aromatization/dearomatization processes of the ligand. A new, dearomatized electron-rich (PNP*)Ir(I) complex 2 (PNP* = deprotonated PNP) activates benzene to form the aromatic (PNP)Ir(I)Ph 4, which upon treatment with CO undergoes a surprising oxidation process to form (PNP*)Ir(III)(H)CO 6, involving proton migration from the ligand "arm" to the metal, with concomitant dearomatization. 4 undergoes stereoselective activation of H2 to exclusively form the trans-dihydride 7, rather than the expected cis dihydride complex. Our evidence, including D-labeling, suggests the possibility that the Ir(I)-Ph complex is transformed to the dearomatized Ir(III)(Ph)(H) (independently prepared at low temperature), which may be the actual intermediate undergoing H2 activation. PMID- 17132003 TI - Solid-phase total synthesis and structure proof of callipeltin B. AB - The cytotoxic, cyclic heptadepsipeptide, natural product callipeltin B was synthesized on a solid-phase support in 15% overall yield. Comparison of the 1H NMR spectra of three synthetic isomers with those of callipeltin B confirmed the configurational reassignment of its threonine residues as d-allothreonine and the assignment of the configuration of its beta-methoxytyrosine residue as (2R,3R). PMID- 17132004 TI - Stereochemical diversity in asymmetric cyclization via memory of chirality. AB - An enantiodivergent asymmetric cyclization of N-Boc-N-omega-bromoalkyl-alpha amino acid derivatives has been developed. With potassium amide bases in DMF, cyclization proceeds with retention of configuration, while inversion of configuration was observed with lithium amide bases in THF. Chirality of the parent amino acids was preserved during enolate formation and cyclization to give aza-cyclic amino acids in up to 98% ee with retention of configuration or inversion of configuration, depending on the reaction conditions. Thus, both enantiomers of cyclic amino acids with a tetrasubstituted stereocenter were prepared in high enantiomeric purity from readily available l-alpha-amino acids. This protocol is also applicable to a spirocyclization and an intramolecular conjugate addition of alpha-amino acid derivatives, giving either of the enantiomers of a diazaspiro compound and a tetrahydroisoquinoline derivative, respectively, in up to 99% ee. PMID- 17132005 TI - Controlling regiochemistry in Negishi carboaluminations. Fine tuning the ligand on zirconium. AB - The species on the zirconocene catalyst is changed from two Cp's to the Brintzinger ligand and catalytic amounts of MAO are used to usually effect a >99% regiocontrol of Negishi carboaluminations of 1-alkynes in toluene. PMID- 17132006 TI - Lithium diisopropylamide solvated by hexamethylphosphoramide: substrate-dependent mechanisms for dehydrobrominations. AB - Lithium diisopropylamide-mediated dehydrobrominations of exo-2-bromonorbornane, 1 bromocyclooctene, and cis-4-bromo-tert-butylcyclohexane were studied in THF solutions and THF solutions with added hexamethylphosphoramide (HMPA). Rate studies reveal a diverse array of mechanisms based on mono-, di-, and trisolvated monomers as well as triple ions. The results are contrasted with analogous eliminations in THF in the absence of HMPA. PMID- 17132007 TI - Particle-wire-tube mechanism for carbon nanotube evolution. AB - The synthesis of carbon nanotubes (CNTs) has been proved to be greatly promoted by vapor metal catalysts, but the fast reaction feature and the required high temperature environment involved in CNT evolution usually make it difficult for an insight into the evolution mechanism. Here, we successfully freeze the synthetic reaction at intermediary stages and observe the detailed morphologies and structures of the obtained intermediates and various objects related to carbon nanotubes. It is unveiled that there is a kindred evolution linkage among carbon nanoparticles, nanowires, and nanotubes in the vapor catalyst-involved synthetic processes: tiny carbon nanoparticles first form from a condensation of gaseous carbon species and then self-assemble into nanowires driven by an anisotropic interaction, and the nanowires finally develop into nanotubes, as a consequence of particle coalescence and structural crystallization. The function of metals is to promote the anisotropic interactions between the nanoparticles and the structural crystallization. An annealing transformation of carbon nanoparticles into nanotubes is also achieved, which gives further evidence for the evolution mechanism. PMID- 17132008 TI - Synthesis and reactivity of azapalladacyclobutanes. AB - N-Sulfonyl aziridines undergo oxidative addition to palladium(0) complexes generated in situ from mixtures of Pd2(dba)3 and 1,10-phenanthroline. The resulting azapalladacyclobutane complexes undergo intramolecular carbopalladation in the presence of copper(I) iodide to afford azapalladabicyclo[3.2.1]octanes. A deuterium-labeling experiment indicates that the oxidative addition proceeds via SN2-type attack of palladium(0) on the less-hindered carbon of the aziridine ring and that alkene insertion occurs in a syn fashion. The azapalladabicyclo[3.2.1]octane complexes undergo oxidative palladium-carbon bond functionalization in the presence of copper(II) bromide. PMID- 17132009 TI - Radical/Ion pair formation in the electrochemical reduction of arene sulfenyl chlorides. AB - Important aspects of the electrochemical reduction of a series of substituted arene sulfenyl chlorides are investigated. A striking change is observed in the reductive cleavage mechanism as a function of the substituent on the aryl ring of the arene sulfenyl chloride. With p-substituted phenyl chlorides a "sticky" dissociative ET mechanism takes place where a concerted ET mechanism leads to the formation of a radical/anion cluster before decomposition. With o-nitropheyl sulfenyl substituted chlorides a stepwise mechanism is observed where through space S...O interactions play an important role stabilizing both the neutral molecules and their reduced forms. Disulfides are generated through a nucleophilic reaction of the two-electron reduction produced anion (arenethiolate) on the parent molecule. The dissociative electron transfer theory, as well as its extension to the case of strong in-cage interactions between the produced fragments, along with the gas phase chemical quantum calculations results helped rationalize both the observed change in the ET mechanism and the occurrence of the "sticky dissociative" ET mechanism. The radical/anion pair interactions have been determined both in solution as well as in gas phase. This study shows that despite the low magnitude of in-cage interactions in acetonitrile as compared to in the gas phase, their existence strongly affects the kinetics of the involved reactions. It also shows that, as expected, these interactions are reinforced by the existence of strong electron withdrawing substituents. PMID- 17132010 TI - An efficient computational method for predicting rotational diffusion tensors of globular proteins using an ellipsoid representation. AB - We propose a new computational method for predicting rotational diffusion properties of proteins in solution. The method is based on the idea of representing protein surface as an ellipsoid shell. In contrast to other existing approaches this method uses principal component analysis of protein surface coordinates, which results in a substantial increase in the computational efficiency of the method. Direct comparison with the experimental data as well as with the recent computational approach (Garcia de la Torre; et al. J. Magn. Reson. 2000, B147, 138-146), based on representation of protein surface as a set of small spherical friction elements, shows that the method proposed here reproduces experimental data with at least the same level of accuracy and precision as the other approach, while being approximately 500 times faster. Using the new method we investigated the effect of hydration layer and protein surface topography on the rotational diffusion properties of a protein. We found that a hydration layer constructed of approximately one monolayer of water molecules smoothens the protein surface and effectively doubles the overall tumbling time. We also calculated the rotational diffusion tensors for a set of 841 protein structures representing the known protein folds. Our analysis suggests that an anisotropic rotational diffusion model is generally required for NMR relaxation data analysis in single-domain proteins, and that the axially symmetric model could be sufficient for these purposes in approximately half of the proteins. PMID- 17132011 TI - A general polymer-based process to prepare mixed metal oxides: the case of Zn1 xMgxO nanoparticles. AB - Nanometer-sized mixed metal oxide (MMO) particles (Zn1-xMgxO) with very precise stoichiometry are prepared employing a polymer-based method. The precursor is formed by loading a polyacrylate with metal ions followed by purification of the polymer metal ion complex via repeated precipitation/redissolution cycles. Calcination of the polymer precursor at 550 degrees C gives particles of the metastable solid solution of the ZnO/MgO system in the composition range (x<0.2 and x>or=0.82). The MMO crystal particles are typically 20-50 nm in diameter. Doping of the ZnO by Mg2+ causes a shrinkage of lattice parameter c. Effects of band gap engineering on the optical band gap are reported. The photoluminescence in the visible is also affected, and its maximum shifts from 2.12 eV (pure ZnO) to 2.32 eV at x=0.21. The crystalline MMO particles start to undergo segregation into hexagonal and cubic phases upon annealing at 800 degrees C. PMID- 17132012 TI - Characterization of manganese(V)-oxo polyoxometalate intermediates and their properties in oxygen-transfer reactions. AB - A manganese(III)-substituted polyoxometalate, [alpha2-P2MnIII(L)W17O61]7- (P2W17MnIII), was studied as an oxidation catalyst using iodopentafluorobenzene bis(tifluoroacetate) (F5PhI(TFAc)2) as a monooxygen donor. Pink P2W17MnIII turns green upon addition of F5PhI(TFAc)2. The 19F NMR spectrum of F5PhI(TFAc)2 with excess P2W17MnIII at -50 degrees C showed the formation of an intermediate attributed to P2W17MnIII-F5PhI(TFAc)2 that disappeared upon warming. The 31P NMR spectra of P2W17MnIII with excess F5PhI(TFAc)2 at -50 and -20 degrees C showed a pair of narrow peaks attributed to a diamagnetic, singlet manganese(V)-oxo species, P2W17MnV=O. An additional broad peak at -10.6 ppm was attributed to both the P2W17MnIII-F5PhI(TFAc)2 complex and a paramagnetic, triplet manganese(V)-oxo species. The electronic structure and reactivity of P2W17MnV=O were modeled by DFT calculations using the analogous Keggin compound, [PMnV=OW11O39]4-. Calculations with a pure functional, UBLYP, showed singlet and triplet ground states of similar energy. Further calculations using both the UBLYP and UB3LYP functionals for epoxidation and hydroxylation of propene showed lowest lying triplet transition states for both transformations, while singlet and quintet transition states were of higher energy. The calculations especially after corrections for the solvent effect indicate that [PMnV=OW11O39]4- should be highly reactive, even more reactive than analogous MnV=O porphyrin species. Kinetic measurements of the reaction of P2W17MnV=O with 1-octene indicated, however, that P2W17MnV=O was less reactive than a MnV=O porphyrin. The experimental enthalpy of activation confirmed that the energy barrier for epoxidation is low, but the highly negative entropy of activation leads to a high free energy of activation. This result originates in our view from the strong solvation of the highly charged polyoxometalate by the polar solvent used and adventitious water. The higher negative charge of the polyoxometalate in the transition versus ground state leads to electrostriction of the solvent molecules and to a loss of degrees of freedom, resulting in a highly negative entropy of activation and slower reactions. PMID- 17132013 TI - Characterization of structure and stability of long telomeric DNA G-quadruplexes. AB - In the current study, we used a combination of gel electrophoresis, circular dichroism, and UV melting analysis to investigate the structure and stability of G-quadruplexes formed by long telomeric DNAs from Oxytricha and human, where the length of the repeat (n)=4 to 12. We found that the Oxytricha telomeric DNAs, which have the sequence (TTTTGGGG)n, folded into intramolecular and intermolecular G-quadruplexes depending on the ionic conditions, whereas human telomeric DNAs, which have the sequence (TTAGGG)n, formed only intramolecular G quadruplexes in all the tested conditions. We further estimated the thermodynamic parameters of the intramolecular G-quadruplex. We found that thermodynamic stabilities of G-quadruplex structures of long telomeric DNAs (n=5 to 12) are mostly independent of sequence length, although telomeric DNAs are more stable when n=4 than when n>or=5. Most importantly, when n is a multiple of four, the change in enthalpy and entropy for G-quadruplex formation increased gradually, demonstrating that the individual G-quadruplex units are composed of four repeats and that the individual units do not interact. Therefore, we propose that the G quadruplexes formed by long telomeric DNAs (n>or=8) are bead-on-a-string structures in which the G-quadruplex units are connected by one TTTT (Oxytricha) or TTA (human) linker. These results should be useful for understanding the structure and function of telomeres and for developing improved therapeutic agents targeting telomeric DNAs. PMID- 17132014 TI - Crystal structure and magnetic properties of FeTe2O5X (X=Cl, Br): a frustrated spin cluster compound with a new Te(IV) coordination polyhedron. AB - A new layered transition metal oxohalide, FeTe2O5ClxBr1-x, has been identified. It crystallizes in the monoclinic space group P21/c. The unit cell for FeTe2O5Br is a = 13.3964(8), b = 6.5966(4), c = 14.2897(6) A, beta=108.118(6) degrees, and Z=8. The layers are built of edge sharing [FeO6] octahedra forming [Fe4O16]20- units that are linked by [Te4O10X2]6- groups. The layers have no net charge and are only weakly connected via van der Waals forces to adjacent layers. There are four crystallographically different Te atoms, and one of them displays a unique [TeO2X] coordination polyhedron (X=Cl, Br). Magnetic susceptibility measurements show a broad maximum typical for 4-spin clusters of coupled Fe(III) ions in the high-spin state. Evidence for magnetic instabilities exists at low temperatures, which have been confirmed with specific heat experiments. A theoretical modeling of the susceptibility concludes a frustration of the intra-tetramer anti ferromagnetic exchange interaction. PMID- 17132015 TI - A modular approach toward regulating the secondary coordination sphere of metal ions: differential dioxygen activation assisted by intramolecular hydrogen bonds. AB - Metal ion function depends on the regulation of properties within the primary and second coordination spheres. An approach toward studying the structure-function relationships within the secondary coordination sphere is to construct a series of synthetic complexes having constant primary spheres but structurally tunable secondary spheres. This was accomplished through the development of hybrid urea carboxamide ligands that provide varying intramolecular hydrogen bond (H-bond) networks proximal to a metal center. Convergent syntheses prepared ligands [(N' tert-butylureayl)-N-ethyl]-bis(N' '-R-carbamoylmethyl)amine (H(4)1R) and bis[(N' tert-butylureayl)-N-ethyl]-(N' '-R-carbamoylmethyl)amine (H(5)2R), where R=isopropyl, cyclopentyl, and (S)-(-)-alpha-methylbenzyl. The ligands with isopropyl groups H(4)1iPr and H(5)2iPr were combined with tris[(N'-tert butylureayl)-N-ethyl]amine (H6buea) and bis(N-isopropylcarbamoylmethyl)amine (H(3)0iPr) to prepare a series of Co(II) complexes with varying H-bond donors. [CoIIH(2)2iPr]- (two H-bond donors), [CoIIH1iPr]- (one H-bond donor), and [CoII0iPr]- (no H-bond donors) have trigonal monopyramidal primary coordination spheres as determined by X-ray diffraction methods. In addition, these complexes have nearly identical optical and EPR properties that are consistent with S=3/2 ground states. Electrochemical studies show a linear spread of 0.23 V in anodic potentials (Epa) with [CoIIH(2)2iPr]- being the most negative at -0.385 V vs [Cp2Fe]+/[Cp2Fe]. The properties of [CoIIH3buea]- (H3buea, tris[(N'-tert butylureaylato)-N-ethyl]aminato that has three H-bond donors) appears to be similar to that of the other complexes based on spectroscopic data. [CoIIH3buea]- and [CoIIH(2)2iPr]- react with 0.5 equiv of dioxygen to afford [CoIIIH3buea(OH)]- and [CoIIIH(2)2iPr(OH)]-. Isotopic labeling studies confirm that dioxygen is the source of the oxygen atom in the hydroxo ligands: [CoIIIH3buea(16OH)]- has a -(O H) band at 3589 cm-1 that shifts to 3579 cm-1 in [CoIIIH3buea(18OH)]-; [CoIIIH(2)2iPr(OH)]- has -(16O-H)=3661 and -(18O-H)=3650 cm-1. [CoIIH1iPr]- does not react with 0.5 equiv of O2; however, treating [CoIIH1iPr]- with excess dioxygen initially produces a species with an X-band EPR signal at g=2.0 that is assigned to a Co-O2 adduct, which is not stable and converts to a species having properties similar to those of the CoIII-OH complexes. Isolation of this hydroxo complex in pure form was complicated by its instability in solution (kint=2.5x10 7 M min-1). Moreover, the stability of the CoIII-OH complexes is correlated with the number of H-bond donors within the secondary coordination sphere; [CoIIIH3buea(OH)]- is stable in solution for days, whereas [CoIIIH(2)2iPr(OH)]- decays with a kint=5.9x10-8 M min-1. The system without any intramolecular H-bond donors [CoII0iPr]- does not react with dioxygen, even when O2 is in excess. These findings indicate a correlation between dioxygen binding/activation and the number of H-bond donors within the secondary coordination sphere of the cobalt complexes. Moreover, the properties of the secondary coordination sphere affect the stability of the CoIII-OH complexes with [CoIIIH3buea(OH)]- being the most stable. We suggest that the greater number of intramolecular H-bonds involving the hydroxo ligand reduces the nucleophilicity of the CoIII-OH unit and reinforces the cavity structure, producing a more constrained microenvironment around the cobalt ion. PMID- 17132016 TI - Short-lived quinonoid species from 5,6-dihydroxyindole dimers en route to eumelanin polymers: integrated chemical, pulse radiolytic, and quantum mechanical investigation. AB - The transient species formed by oxidation of three dimers of 5,6-dihydroxyindole (1), a major building block of the natural biopolymer eumelanin, have been investigated. Pulse radiolytic oxidation of 5,5',6,6'-tetrahydroxy-2,4'-biindolyl (3) and 5,5',6,6'-tetrahydroxy-2,7'-biindolyl (4) led to semiquinones absorbing around 450 nm, which decayed with second-order kinetics (2k=2.8x10(9) and 1.4x10(9) M-1 s-1, respectively) to give the corresponding quinones (500-550 nm). 5,5',6, 6'-Tetrahydroxy-2,2'-biindolyl (2), on the other hand, furnished a semiquinone (lamdamax=480 nm) which disproportionated at a comparable rate (2k=3x10(9) M-1 s-1) to give a relatively stable quinone (lamdamax=570 nm). A quantum mechanical investigation of o-quinone, quinonimine, and quinone methide structures of 2-4 suggested that oxidized 2-4 exist mainly as 2-substituted extended quinone methide tautomers. Finally, an oxidation product of 3 was isolated for the first time and was formulated as the hydroxylated derivative 5 arising conceivably by the addition of water to the quinone methide intermediate predicted by theoretical analysis. Overall, these results suggest that the oxidation chemistry of biindolyls 2-4 differs significantly from that of the parent 1, whereby caution must be exercised before concepts that apply strictly to the mode of coupling of 1 are extended to higher oligomers. PMID- 17132017 TI - Energetic determinants of oligomeric state specificity in coiled coils. AB - The coiled coil is one of the simplest and best-studied protein structural motifs, consisting of two to five helices wound around each other. Empirical rules have been established on the tendency of different core sequences to form a certain oligomeric state but the physical forces behind this specificity are unclear. In this work, we model four sequences onto the structures of dimeric, trimeric, tetrameric, and pentameric coiled coils. We first examine the ability of an effective energy function (EEF1.1) to discriminate the correct oligomeric state for a given sequence. We find that inclusion of the translational, rotational, and side-chain conformational entropy is necessary for discriminating the native structures from their misassembled counterparts. The decomposition of the effective energy into residue contributions yields theoretical values for the oligomeric propensity of different residue types at different heptad positions. We find that certain calculated residue propensities are general and consistent with existing rules. For example, leucine at d favors dimers, leucine at a favors tetramers or pentamers, and isoleucine at a favors trimers. Other residue propensities are sequence context dependent. For example, glutamine at d favors trimers in one context and pentamers in another. Charged residues at e and g positions usually destabilize higher oligomers due to higher desolvation. Nonpolar residues at these positions confer pentamer specificity when combined with certain residues at positions a and d. Specifically, the pair Leua-Alag' or the inverse was found to stabilize the pentamer. The small energy gap between the native and misfolded counterparts explains why a few mutations at the core sites are sufficient to induce a change in the oligomeric state of these peptides. A large number of possible experiments are suggested by these results. PMID- 17132018 TI - (n,m) Abundance evaluation of single-walled carbon nanotubes by fluorescence and absorption spectroscopy. AB - A methodology that takes into account the (n,m) structure of single-walled carbon nanotubes (SWNTs), through an exciton-exciton resonance model and an electron phonon interaction model, was employed in order to evaluate the semiconducting (n,m) abundance of two SWNT samples (i.e., Co-MCM-41 and HiPco). This was based on photoluminescence and near-infrared absorption data obtained on aqueous suspensions of individually dispersed SWNTs. In the absence of known (n,m) abundance SWNT samples, we resorted to determining the diameter distribution curves for both samples, which were found to obey an unsymmetrical log-normal distribution, typical for vapor-phase particle growth. Using this log-normal function, we reconstructed the near-infrared E S11 absorption spectrum of the narrow diameter distribution Co-MCM-41 SWNT sample, which in turn enabled us to assess the predictions of these two theoretical models. High spectral reconstruction accuracy was obtained from the electron-phonon interaction model when considering (11,0) and (10,0) zigzag nanotubes, along with (n,m) line widths inversely proportional to their extinction coefficients. PMID- 17132019 TI - Zinspy sensors with enhanced dynamic range for imaging neuronal cell zinc uptake and mobilization. AB - Thiophene moieties were incorporated into previously described Zinspy (ZS) fluorescent Zn(II) sensor motifs (Nolan, E. M.; Lippard, S. J. Inorg. Chem. 2004, 43, 8310-8317) to provide enhanced fluorescence properties, low-micromolar dissociation constants for Zn(II), and improved Zn(II) selectivity. Halogenation of the xanthenone and benzoate moieties of the fluorescein platform systematically modulates the excitation and emission profiles, pH-dependent fluorescence, Zn(II) affinity, and Zn(II) complexation rates, offering a general strategy for tuning multiple properties of xanthenone-based metal ion sensors. Extensive biological studies in cultured cells and primary neuronal cultures demonstrate 2-{6-hydroxy-3-oxo-4,5-bis[(pyridin-2-ylmethylthiophen-2 ylmethylamino)methyl]-3H-xanthen-9-yl}benzoic acid (ZS5) to be a versatile imaging tool for detecting Zn(II) in vivo. ZS5 localizes to the mitochondria of HeLa cells and allows visualization of glutamate-mediated Zn(II) uptake in dendrites and Zn(II) release resulting from nitrosative stress in neurons. PMID- 17132020 TI - Novobiocin: redesigning a DNA gyrase inhibitor for selective inhibition of hsp90. AB - Novobiocin is a member of the coumermycin family of antibiotics and is a well established inhibitor of DNA gyrase. Recent studies have shown that novobiocin binds to a previously unrecognized ATP-binding site at the C-terminus of Hsp90 and induces degradation of Hsp90-dependent client proteins at approximately 700 microM. In an effort to develop more efficacious inhibitors of the C-terminal binding site, a library of novobiocin analogues was prepared and initial structure-activity relationships revealed. These data suggested that the 4 hydroxy moiety of the coumarin ring and the 3'-carbamate of the noviose appendage were detrimental to Hsp90 inhibitory activity. In an effort to confirm these findings, 4-deshydroxy novobiocin (DHN1) and 3'-descarbamoyl-4 deshydroxynovobiocin (DHN2) were prepared and evaluated against Hsp90. Both compounds were significantly more potent than the natural product, and DHN2 proved to be more active than DHN1. In an effort to determine whether these moieties are important for DNA gyrase inhibition, these compounds were tested for their ability to inhibit DNA gyrase and found to exhibit significant reduction in gyrase activity. Thus, we have established the first set of compounds that clearly differentiate between the C-terminus of Hsp90 and DNA gyrase, converted a well-established gyrase inhibitor into a selective Hsp90 inhibitor, and confirmed essential structure-activity relationships for the coumermycin family of antibiotics. PMID- 17132021 TI - Self-organized monolayer of meso-tetradodecylporphyrin coordinated to Au(111). AB - The structure of molecular monolayers formed at the interface between atomically flat surfaces and a solution of free-base meso-tetradodecylporphyrins (H2Ps) was examined by scanning tunneling microscopy (STM) at the liquid/solid interface. On the surface of graphite (HOPG), H2Ps form a well-ordered monolayer characterized by an oblique unit cell. On Au(111), H2Ps form a self-organized monolayer comprised of two distinct domain types. In both types of domains, the density of the porphyrin cores is increased in comparison to the arrangement observed on HOPG. Also, high-resolution STM images reveal that, in contrast to what is observed on HOPG, physisorption on Au(111) induces a distortion of the porphyrin macrocycle out of planarity. By using X-ray photoelectron spectroscopy, we demonstrate that this is likely to be due to the coordination of the lone pairs of the iminic (-C=N-) nitrogen atoms of the porphyrin macrocycle to Au(111). PMID- 17132022 TI - Immobilization of ligands with precise control of density to electroactive surfaces. AB - We report a broadly applicable surface chemistry methodology to immobilize ligands, proteins, and cells to an electroactive substrate with precise control of ligand density. This strategy is based on the coupling of soluble aminooxy terminated ligands with an electroactive quinone terminated monolayer. The surface chemistry product oxime is also redox active but at a different potential and therefore allows for real-time monitoring of the immobilization reaction. Only the quinone form of the immobilized redox pair is reactive with soluble aminooxy groups, which allows for the determination of the yield of reaction, the ability to immobilize multiple ligands at controlled densities, and the in-situ modulation of ligand activity. We demonstrate this methodology by using cyclic voltammetry to characterize the kinetics of a model interfacial reaction with aminooxy acetic acid. We also demonstrate the synthetic flexibility and utility of this method for biospecific interactions by installing aminooxy terminated FLAG peptides and characterizing their binding to soluble anti-FLAG with surface plasmon resonance spectroscopy. We further show this methodology is compatible with microarray technology by printing rhodamine-oxyamine in various size spots and characterizing the yield within the spots by cyclic voltammetry. We also show this methodology is compatible with cell culture conditions and fluorescent microscopy technology for cell biological studies. Arraying RGD-oxyamine peptides on these substrates allows for bio-specific adhesion of Swiss 3T3 Fibroblasts. PMID- 17132029 TI - To retain or remove user fees?: reflections on the current debate in low- and middle-income countries. AB - Many low- and middle-income countries continue to search for better ways of financing their health systems. Common to many of these systems are problems of inadequate resource mobilisation, as well as inefficient and inequitable use of existing resources. The poor and other vulnerable groups who need healthcare the most are also the most affected by these shortcomings. In particular, these groups have a high reliance on user fees and other out-of-pocket expenditures on health which are both impoverishing and provide a financial barrier to care. It is within this context, and in light of recent policy initiatives on user fee removal, that a debate on the role of user fees in health financing systems has recently returned. This paper provides some reflections on the recent user fees debate, drawing from the evidence presented and subsequent discussions at a recent UNICEF consultation on user fees in the health sector, and relates the debate to the wider issue of access to adequate healthcare. It is argued that, from the wealth of evidence on user fees and other health system reforms, a broad consensus is emerging. First, user fees are an important barrier to accessing health services, especially for poor people. They also negatively impact on adherence to long-term expensive treatments. However, this is offset to some extent by potentially positive impacts on quality. Secondly, user fees are not the only barrier that the poor face. As well as other cost barriers, a number of quality, information and cultural barriers must also be overcome before the poor can access adequate health services. Thirdly, initial evidence on fee abolition in Uganda suggests that this policy has improved access to outpatient services for the poor. For this to be sustainable and effective in reaching the poor, fee removal needs to be part of a broader package of reforms that includes increased budgets to offset lost fee revenue (as was the case in Uganda). Fourthly, implementation matters: if fees are to be abolished, this needs clear communication with a broad stakeholder buy-in, careful monitoring to ensure that official fees are not replaced by informal fees, and appropriate management of the alternative financing mechanisms that are replacing user fees. Fifthly, context is crucial. For instance, immediate fee removal in Cambodia would be inappropriate, given that fees replaced irregular and often high informal fees. In this context, equity funds and eventual expansion of health insurance are perhaps more viable policy options. Conversely, in countries where user fees have had significant adverse effects on access and generated only limited benefits, fee abolition is probably a more attractive policy option. Removing user fees has the potential to improve access to health services, especially for the poor, but it is not appropriate in all contexts. Analysis should move on from broad evaluations of user fees towards exploring how best to dismantle the multiple barriers to access in specific contexts. PMID- 17132030 TI - Modelling the costs and outcomes of changing rates of screening for alcohol misuse by GPs in the Australian context. AB - AIM: To assess the relative cost effectiveness of four strategies (academic detailing, computerised reminder systems, target payments and interactive continuing medical education) to increase the provision of screening and brief interventions by Australian GPs with the ultimate goal of decreasing risky alcohol consumption among their patients. METHODS: This project used a modelling approach to combine information on the effectiveness and costs of four separate strategies to change GP behaviours to estimate their relative cost effectiveness. RESULTS: The computerised reminder system and academic detailing appear most effective in achieving a decrease in the number of standard drinks consumed by risky drinkers. CONCLUSION: Regardless of the assumptions made, the targeted payment strategy appeared to be the least cost-effective method to achieve a decrease in risky alcohol consumption while the other three strategies appear reasonably comparable. PMID- 17132031 TI - Community-based youth tobacco control interventions: cost effectiveness of the Full Court Press project. AB - BACKGROUND: We evaluated the impact of a community-based tobacco control project that was implemented in the city of Tucson, Arizona, USA, between 1996 and 2001. AIM: The project's goal was to reduce the prevalence of youth smoking through change in social norms at schools and in communities and workplaces. As is often the case, these community-based health promotion interventions were implemented in conjunction with other broader programmes, in this case implemented on the state level. METHOD: Taking into account state level interventions as well as changes in sociodemographic and economic environment over the course of the project (e.g. increases in cigarette prices), we measure the net effect of the intervention in terms of the number of people who quit or did not initiate smoking and by the discounted life-years gained. To establish the value of investing into community-based intervention, we calculated the real discounted cost per quit and per life-year gained of 3789 US dollars and 3942 US dollars, respectively. These compare favourably with the real cost per quit of 4270 US dollars when implementing the 1996 US Clinical Practice Guideline for smoking cessation but exceed the real cost of 2923 US dollars per discounted life-year gained when following the guideline. RESULTS: A sensitivity analysis that assumed 5% programme persistence (i.e. 5% of the programme's impact would last forever in the absence of future funding for the programme), one-third would relapse and that one-third of those who quit may have quit smoking even without the programme, suggested a lower cost per discounted life-year saved of 3476 US dollars. The cost effectiveness of this project compares favourably with other tobacco control interventions. CONCLUSION: Despite its relatively small target group, this community-based intervention was cost effective. PMID- 17132032 TI - Comparing patient access to pharmaceuticals in the UK and US. AB - BACKGROUND: The debate on access to new drugs has focused on the time lag between applications for approval and granting of marketing authorisation. This delay was identified as the first barrier with respect to patient access to new drugs, encompassing the hurdles of safety, efficacy and quality. Additional barriers have since been identified. These pertain to reimbursement and pricing of approved drugs, the so-called fourth and fifth hurdles. METHODS: We reviewed 38 National Institute for Health and Clinical Excellence (NICE) guidance appraisals carried out between April 1999 and April 2005. These appraisals included 71 recently approved drugs considered to have either high clinical or cost impact. For each drug we first determined its marketing approval date by the British Medicines Healthcare Products Agency (MHRA) or European Medicines Evaluation Agency (EMEA). Secondly, we determined if each drug was approved by the US FDA for marketing and, if so, the date when it was approved. Thirdly, we considered whether and when each drug was recommended for reimbursement and use by NICE, and whether conditions of reimbursement applied. Fourthly, for the subset of FDA approved drugs, we examined formulary placement, cost sharing and conditions of reimbursement on three-tier formularies used by seven leading US third-party payers serving Medicare beneficiaries. Fifthly, we reviewed each NICE recommendation to determine if cost-effectiveness data were referred to either in the appraisal documentation or in the final recommendation. Sixthly, we asked a spokesperson from each US payer whether cost-effectiveness assessments or rebates played a role in determining formulary placement of drugs in our sample, and whether there was a lag between marketing approval and reimbursement for any of the covered drugs. RESULTS: Of the 71 drugs contained in 38 NICE guidance appraisals, the US FDA approved 64. On average, the subset of 64 drugs received marketing authorisation in the US prior to the UK. On average, US plans covered 87% of the 64 drugs, the same percentage of drugs recommended for NHS reimbursement and use. Cost sharing in the US was significantly higher than in the UK, with wider variation across plans. On average, drugs covered in the US had fewer conditions of reimbursement (15%) than the percentage of drugs given conditions by NICE (46%). US plans were quicker to decide to reimburse drugs following marketing approval than NICE. CONCLUSIONS: The US provides faster, more flexible access to most, but not all, of the UK-approved pharmaceuticals in our sample. However, US patients have higher cost sharing than the UK and coverage is less evenly spread across the population. From a policy perspective, our study findings confirm the need to bolster the NICE fast-track initiative to decrease the amount of time it takes to appraise certain new pharmaceuticals. Also, the study findings point to the need in the US for careful monitoring of plan compliance with regulations pertaining to the Medicare drug benefit, particularly with respect to formulary restrictions and limits on cost sharing. PMID- 17132033 TI - Patients' preferences for healthcare system reforms in Hungary: a conjoint analysis. AB - OBJECTIVES: To illustrate how conjoint analysis can be used to identify patient preferences for healthcare policies, and to measure preferences for healthcare reforms in Hungary. DATA SOURCE/STUDY SETTING: Data was collected via a mail based survey and a direct survey administered in a rheumatology out-patient centre in Flor Ferenc County Hospital, Budapest, Hungary (n = 86). STUDY DESIGN: We designed and administered a conjoint analysis to the study population. Attributes and attribute levels were developed on the basis of key informant interviews and a literature review. Additional demographic, occupation and healthcare utilisation data were also collected using surveys. A mixed effects linear probability model was estimated holding respondent characteristics constant and correcting for clustering. DATA COLLECTION: Conjoint analysis questionnaires were administered by a physician to 50 consecutive rheumatology patients in a clinic and an additional 36 were mailed by post. PRINCIPAL FINDINGS: The response rate for the physician-administered survey was 98% (but 18% of these were excluded for inconsistent preferences) and 53% for the mail survey, leaving a final sample of 59. Regression results (R2 = 56.8%) indicated that patients preferred a health system that was not cost constrained (p = 0.003), was based on solidarity (p < 0.001) and where patients were empowered (p = 0.024). Further, they would choose a system with no choice of provider to avoid co-payments (p = 0.005). CONCLUSIONS: This study demonstrates that patients have clear preferences for healthcare system policy. In order to develop evidence based healthcare policy and to empower patients in the healthcare system, methods such as conjoint analysis offer a simple yet theoretically grounded basis for policy making. PMID- 17132034 TI - Juggling and struggling: a preliminary work-life study of mothers with adolescents who have developmental disabilities. AB - A focus group study was conducted to develop an understanding of the experiences of mothers who are trying to balance employment with caring for an adolescent with developmental disabilities. Mothers reported facing considerable difficulties balancing work and caregiving responsibilities because support services rapidly declined when their child reached adolescence. Service cuts were related to the fact that adolescents are expected to be able to care for themselves, despite the fact that for many adolescents with disabilities, this is not possible. The mothers also reported that the preponderance of the responsibility for arranging care for their children was theirs and was not shouldered by their partners. Policy implications are discussed. PMID- 17132035 TI - Consumer-directed supports: economic, health, and social outcomes for families. AB - The impact of a consumer-directed support program on family caregivers of adults with developmental disabilities was explored. Economic, health, and social outcomes were compared between families in the program and families on the waiting list for the program. Caregivers of adults in the program reported fewer out-of-pocket disability expenses, greater access to health care, engagement in more social activities, and greater leisure satisfaction. There also appeared to be greater impacts on lower income families; these caregivers reported better mental health and access to health care than did similar caregivers on the waiting list. PMID- 17132036 TI - Longitudinal frequency and stability of family contact in institutional and community living. AB - We examined the frequency and stability of family contact with long-term institutional residents during a major deinstitutionalization project. Movers relocated to community accommodation between Assessments 1 (baseline) and 2. Stayers remained institutionalized. We investigated family contact longitudinally over four annual assessments. There was no significant between-group difference in frequency of family contact at baseline, but, subsequently, movers had more frequent family contact than did stayers. There was a significant decline over time in the proportion of stayers with family contact and a significant increase in movers with family contact, with a marked resumption of contact by movers during the first year of community living. Carefully supporting family (re)involvement during the relocation process may be associated with stable, long term family contact. PMID- 17132037 TI - The "big bang" theory and Down syndrome. PMID- 17132038 TI - "Uncle Sam needs you" or does he? Intellectual disabilities and lessons from the "great wars". PMID- 17132039 TI - Incident management, organizational culture, and honest communications. PMID- 17132040 TI - Community literacy and friendship model for people with intellectual disabilities. PMID- 17132041 TI - Twenty-year retrospective on proposals to eliminate the "institutional bias" in Medicaid for persons with ID/DD. PMID- 17132042 TI - Dosimetric and spectroradiometric investigations of glass filtered solar UV. AB - The aims of this paper were to investigate how glass-filtered UV irradiances vary with glass thickness, lamination of the glass and the effect of solar zenith angle (SZA), and to measure the glass-filtered UV exposures to different receiving planes with a newly developed UVA dosimeter. Spectroradiometric and dosimetric techniques were employed in the experimental approach. The percentage of the glass-filtered solar UV compared to the unfiltered UV ranged from 59% to 70% and was influenced to a small extent by the glass thickness and the SZA. The laminated glass transmitted 11 to 12% and the windscreen glass transmitted 2.5 2.6%. The influence of the SZA was less for the thicker glass than it was for the thinner glass. The change in transmission was less than 14% for the SZA between 48 degrees and 71 degrees. There was negligible influence due to the SZA on the glass-transmitted UV of the laminated and windscreen glass. The influence of the glass thickness in the range of 2-6 mm on the percentage transmission was less than 16%. The influences of the glass thickness and the SZA on the glass transmitted UV have been incorporated in the use of a UVA dosimeter for the glass transmitted UV exposures. The UVA dosimeter was employed in the field to measure the glass-filtered UV exposures to different receiving planes. The UVA dosimeter reported has the potential for personal solar UVA exposure measurements. PMID- 17132043 TI - Integrated optical switching based on the protein bacteriorhodopsin. AB - According to our earlier pioneering study, a dry film containing native bacteriorhodopsin (bR) shows unique nonlinear optical properties (refractive index change, controllable by light of different colors, greater than 2 x 10(-3)) that are in many respects superior to those of the materials presently applied in integrated optics. Here, we report on the first integrated optical application based on a miniature Mach-Zehnder interferometer (see Figs. 1 and 2) demonstrating a real switching effect by bR (efficiency higher than 90%) due to the M-state. Our results also imply that the refractive index change of the K state (9 x 10(-4)) is high enough for fast switching. PMID- 17132044 TI - N-terminal and C-terminal domains of arrestin both contribute in binding to rhodopsin. AB - Visual arrestin terminates the signal amplification cascade in photoreceptor cells by blocking the interaction of light activated phosphorylated rhodopsin with the G-protein transducin. Although crystal structures of arrestin and rhodopsin are available, it is still unknown how the complex of the two proteins is formed. To investigate the interaction sites of arrestin with rhodopsin various surface regions of recombinant arrestin were sterically blocked by different numbers of fluorophores (Alexa 633). The binding was recorded by time resolved light scattering. To accomplish site-specific shielding of protein regions, in a first step all three wild-type cysteines were replaced by alanines. Nevertheless, regarding the magnitude and specificity of rhodopsin binding, the protein is still fully active. In a second step, new cysteines were introduced at selected sites to allow covalent binding of fluorophores. Upon attachment of Alexa 633 to the recombinant cysteines we observed that these bulky labels residing in the concave area of either the N- or the C-terminal domain do not perturb the activity of arrestin. By simultaneously modifying both domains with one Alexa 633 the binding capacity was reduced. The presence of two Alexa 633 molecules in each domain prevented binding of rhodopsin to arrestin. This observation indicates that both concave sites participate in binding. PMID- 17132045 TI - How behavioral constraints may determine optimal sensory representations. AB - The sensory-triggered activity of a neuron is typically characterized in terms of a tuning curve, which describes the neuron's average response as a function of a parameter that characterizes a physical stimulus. What determines the shapes of tuning curves in a neuronal population? Previous theoretical studies and related experiments suggest that many response characteristics of sensory neurons are optimal for encoding stimulus-related information. This notion, however, does not explain the two general types of tuning profiles that are commonly observed: unimodal and monotonic. Here I quantify the efficacy of a set of tuning curves according to the possible downstream motor responses that can be constructed from them. Curves that are optimal in this sense may have monotonic or nonmonotonic profiles, where the proportion of monotonic curves and the optimal tuning-curve width depend on the general properties of the target downstream functions. This dependence explains intriguing features of visual cells that are sensitive to binocular disparity and of neurons tuned to echo delay in bats. The numerical results suggest that optimal sensory tuning curves are shaped not only by stimulus statistics and signal-to-noise properties but also according to their impact on downstream neural circuits and, ultimately, on behavior. PMID- 17132046 TI - Genomic and metabolic studies of the impact of probiotics on a model gut symbiont and host. AB - Probiotics are deliberately ingested preparations of live bacterial species that confer health benefits on the host. Many of these species are associated with the fermentation of dairy products. Despite their increasing use, the molecular details of the impact of various probiotic preparations on resident members of the gut microbiota and the host are generally lacking. To address this issue, we colonized germ-free mice with Bacteroides thetaiotaomicron, a prominent component of the adult human gut microbiota, and Bifidobacterium longum, a minor member but a commonly used probiotic. Simultaneous whole genome transcriptional profiling of both bacterial species in their gut habitat and of the intestinal epithelium, combined with mass-spectrometric analysis of habitat-associated carbohydrates, revealed that the presence of B. longum elicits an expansion in the diversity of polysaccharides targeted for degradation by B. thetaiotaomicron (e.g., mannose- and xylose-containing glycans), and induces host genes involved in innate immunity. Although the overall transcriptome expressed by B. thetaiotaomicron when it encounters B. longum in the cecum is dependent upon the genetic background of the mouse (as assessed by a mixed analysis of variance [ANOVA] model of co-colonization experiments performed in NMRI and C57BL/6J animals), B. thetaiotaomicron's expanded capacity to utilize polysaccharides occurs independently of host genotype, and is also observed with a fermented dairy product-associated strain, Lactobacillus casei. This gnotobiotic mouse model provides a controlled case study of how a resident symbiont and a probiotic species adapt their substrate utilization in response to one another, and illustrates both the generality and specificity of the relationship between a host, a component of its microbiota, and intentionally consumed microbial species. PMID- 17132047 TI - Rarity value and species extinction: the anthropogenic Allee effect. AB - Standard economic theory predicts that exploitation alone is unlikely to result in species extinction because of the escalating costs of finding the last individuals of a declining species. We argue that the human predisposition to place exaggerated value on rarity fuels disproportionate exploitation of rare species, rendering them even rarer and thus more desirable, ultimately leading them into an extinction vortex. Here we present a simple mathematical model and various empirical examples to show how the value attributed to rarity in some human activities could precipitate the extinction of rare species-a concept that we term the anthropogenic Allee effect. The alarming finding that human perception of rarity can precipitate species extinction has serious implications for the conservation of species that are rare or that may become so, be they charismatic and emblematic or simply likely to become fashionable for certain activities. PMID- 17132048 TI - Drosophila NMNAT maintains neural integrity independent of its NAD synthesis activity. AB - Wallerian degeneration refers to a loss of the distal part of an axon after nerve injury. Wallerian degeneration slow (Wld(s)) mice overexpress a chimeric protein containing the NAD synthase NMNAT (nicotinamide mononucleotide adenylyltransferase 1) and exhibit a delay in axonal degeneration. Currently, conflicting evidence raises questions as to whether NMNAT is the protecting factor and whether its enzymatic activity is required for such a possible function. Importantly, the link between nmnat and axon degeneration is at present solely based on overexpression studies of enzymatically active protein. Here we use the visual system of Drosophila as a model system to address these issues. We have isolated the first nmnat mutations in a multicellular organism in a forward genetic screen for synapse malfunction in Drosophila. Loss of nmnat causes a rapid and severe neurodegeneration that can be attenuated by blocking neuronal activity. Furthermore, in vivo neuronal expression of mutated nmnat shows that enzymatically inactive NMNAT protein retains strong neuroprotective effects and rescues the degeneration phenotype caused by loss of nmnat. Our data indicate an NAD-independent requirement of NMNAT for maintaining neuronal integrity that can be exploited to protect neurons from neuronal activity-induced degeneration by overexpression of the protein. PMID- 17132049 TI - Autophagy counterbalances endoplasmic reticulum expansion during the unfolded protein response. AB - The protein folding capacity of the endoplasmic reticulum (ER) is regulated by the unfolded protein response (UPR). The UPR senses unfolded proteins in the ER lumen and transmits that information to the cell nucleus, where it drives a transcriptional program that is tailored to re-establish homeostasis. Using thin section electron microscopy, we found that yeast cells expand their ER volume at least 5-fold under UPR-inducing conditions. Surprisingly, we discovered that ER proliferation is accompanied by the formation of autophagosome-like structures that are densely and selectively packed with membrane stacks derived from the UPR expanded ER. In analogy to pexophagy and mitophagy, which are autophagic processes that selectively sequester and degrade peroxisomes and mitochondria, the ER-specific autophagic process described utilizes several autophagy genes: they are induced by the UPR and are essential for the survival of cells subjected to severe ER stress. Intriguingly, cell survival does not require vacuolar proteases, indicating that ER sequestration into autophagosome-like structures, rather than their degradation, is the important step. Selective ER sequestration may help cells to maintain a new steady-state level of ER abundance even in the face of continuously accumulating unfolded proteins. PMID- 17132050 TI - An unexpected sequence of events: mismatch detection in the human hippocampus. AB - The ability to identify and react to novelty within the environment is fundamental to survival. Computational models emphasize the potential role of the hippocampus in novelty detection, its unique anatomical circuitry making it ideally suited to act as a comparator between past and present experience. The hippocampus, therefore, is viewed to detect associative mismatches between what is expected based on retrieval of past experience and current sensory input. However, direct evidence that the human hippocampus performs such operations is lacking. We explored brain responses to novel sequences of objects using functional magnetic resonance imaging (fMRI), while subjects performed an incidental target detection task. Our results demonstrate that hippocampal activation was maximal when prior predictions concerning which object would appear next in a sequence were violated by sensory reality. In so doing, we establish the biological reality of associative match-mismatch computations within the human hippocampus, a process widely held to play a cardinal role in novelty detection. Our results also suggest that the hippocampus may generate predictions about how future events will unfold, and critically detect when these expectancies are violated, even when task demands do not require it. The present study also offers broader insights into the nature of essential computations carried out by the hippocampus, which may also underpin its unique contribution to episodic memory. PMID- 17132051 TI - Widespread discordance of gene trees with species tree in Drosophila: evidence for incomplete lineage sorting. AB - The phylogenetic relationship of the now fully sequenced species Drosophila erecta and D. yakuba with respect to the D. melanogaster species complex has been a subject of controversy. All three possible groupings of the species have been reported in the past, though recent multi-gene studies suggest that D. erecta and D. yakuba are sister species. Using the whole genomes of each of these species as well as the four other fully sequenced species in the subgenus Sophophora, we set out to investigate the placement of D. erecta and D. yakuba in the D. melanogaster species group and to understand the cause of the past incongruence. Though we find that the phylogeny grouping D. erecta and D. yakuba together is the best supported, we also find widespread incongruence in nucleotide and amino acid substitutions, insertions and deletions, and gene trees. The time inferred to span the two key speciation events is short enough that under the coalescent model, the incongruence could be the result of incomplete lineage sorting. Consistent with the lineage-sorting hypothesis, substitutions supporting the same tree were spatially clustered. Support for the different trees was found to be linked to recombination such that adjacent genes support the same tree most often in regions of low recombination and substitutions supporting the same tree are most enriched roughly on the same scale as linkage disequilibrium, also consistent with lineage sorting. The incongruence was found to be statistically significant and robust to model and species choice. No systematic biases were found. We conclude that phylogenetic incongruence in the D. melanogaster species complex is the result, at least in part, of incomplete lineage sorting. Incomplete lineage sorting will likely cause phylogenetic incongruence in many comparative genomics datasets. Methods to infer the correct species tree, the history of every base in the genome, and comparative methods that control for and/or utilize this information will be valuable advancements for the field of comparative genomics. PMID- 17132052 TI - Projections of global mortality and burden of disease from 2002 to 2030. AB - BACKGROUND: Global and regional projections of mortality and burden of disease by cause for the years 2000, 2010, and 2030 were published by Murray and Lopez in 1996 as part of the Global Burden of Disease project. These projections, which are based on 1990 data, continue to be widely quoted, although they are substantially outdated; in particular, they substantially underestimated the spread of HIV/AIDS. To address the widespread demand for information on likely future trends in global health, and thereby to support international health policy and priority setting, we have prepared new projections of mortality and burden of disease to 2030 starting from World Health Organization estimates of mortality and burden of disease for 2002. This paper describes the methods, assumptions, input data, and results. METHODS AND FINDINGS: Relatively simple models were used to project future health trends under three scenarios-baseline, optimistic, and pessimistic-based largely on projections of economic and social development, and using the historically observed relationships of these with cause-specific mortality rates. Data inputs have been updated to take account of the greater availability of death registration data and the latest available projections for HIV/AIDS, income, human capital, tobacco smoking, body mass index, and other inputs. In all three scenarios there is a dramatic shift in the distribution of deaths from younger to older ages and from communicable, maternal, perinatal, and nutritional causes to noncommunicable disease causes. The risk of death for children younger than 5 y is projected to fall by nearly 50% in the baseline scenario between 2002 and 2030. The proportion of deaths due to noncommunicable disease is projected to rise from 59% in 2002 to 69% in 2030. Global HIV/AIDS deaths are projected to rise from 2.8 million in 2002 to 6.5 million in 2030 under the baseline scenario, which assumes coverage with antiretroviral drugs reaches 80% by 2012. Under the optimistic scenario, which also assumes increased prevention activity, HIV/AIDS deaths are projected to drop to 3.7 million in 2030. Total tobacco-attributable deaths are projected to rise from 5.4 million in 2005 to 6.4 million in 2015 and 8.3 million in 2030 under our baseline scenario. Tobacco is projected to kill 50% more people in 2015 than HIV/AIDS, and to be responsible for 10% of all deaths globally. The three leading causes of burden of disease in 2030 are projected to include HIV/AIDS, unipolar depressive disorders, and ischaemic heart disease in the baseline and pessimistic scenarios. Road traffic accidents are the fourth leading cause in the baseline scenario, and the third leading cause ahead of ischaemic heart disease in the optimistic scenario. Under the baseline scenario, HIV/AIDS becomes the leading cause of burden of disease in middle- and low-income countries by 2015. CONCLUSIONS: These projections represent a set of three visions of the future for population health, based on certain explicit assumptions. Despite the wide uncertainty ranges around future projections, they enable us to appreciate better the implications for health and health policy of currently observed trends, and the likely impact of fairly certain future trends, such as the ageing of the population, the continued spread of HIV/AIDS in many regions, and the continuation of the epidemiological transition in developing countries. The results depend strongly on the assumption that future mortality trends in poor countries will have a relationship to economic and social development similar to those that have occurred in the higher-income countries. PMID- 17132053 TI - Population pharmacokinetics of artesunate and dihydroartemisinin following intra rectal dosing of artesunate in malaria patients. AB - BACKGROUND: Intra-rectal artesunate has been developed as a potentially life saving treatment of severe malaria in rural village settings where administration of parenteral antimalarial drugs is not possible. We studied the population pharmacokinetics of intra-rectal artesunate and the relationship with parasitological responses in patients with moderately severe falciparum malaria. METHODS AND FINDINGS: Adults and children in Africa and Southeast Asia with moderately severe malaria were recruited in two Phase II studies (12 adults from Southeast Asia and 11 children from Africa) with intensive sampling protocols, and three Phase III studies (44 children from Southeast Asia, and 86 children and 26 adults from Africa) with sparse sampling. All patients received 10 mg/kg artesunate as a single intra-rectal dose of suppositories. Venous blood samples were taken during a period of 24 h following dosing. Plasma artesunate and dihydroartemisinin (DHA, the main biologically active metabolite) concentrations were measured by high-performance liquid chromatography with electrochemical detection. The pharmacokinetic properties of DHA were determined using nonlinear mixed-effects modelling. Artesunate is rapidly hydrolysed in vivo to DHA, and this contributes the majority of antimalarial activity. For DHA, a one compartment model assuming complete conversion from artesunate and first-order appearance and elimination kinetics gave the best fit to the data. The mean population estimate of apparent clearance (CL/F) was 2.64 (l/kg/h) with 66% inter individual variability. The apparent volume of distribution (V/F) was 2.75 (l/kg) with 96% inter-individual variability. The estimated DHA population mean elimination half-life was 43 min. Gender was associated with increased mean CL/F by 1.14 (95% CI: 0.36-1.92) (l/kg/h) for a male compared with a female, and weight was positively associated with V/F. Larger V/Fs were observed for the patients requiring early rescue treatment compared with the remainder, independent of any confounders. No associations between the parasitological responses and the posterior individual estimates of V/F, CL/F, and AUC0-6h were observed. CONCLUSIONS: The pharmacokinetic properties of DHA were affected only by gender and body weight. Patients with the lowest area under the DHA concentration curve did not have slower parasite clearance, suggesting that rectal artesunate is well absorbed in most patients with moderately severe malaria. However, a number of modelling assumptions were required due to the large intra- and inter-individual variability of the DHA concentrations. PMID- 17132054 TI - Putting it together: AIDS and the millennium development goals. PMID- 17132055 TI - Fluoxetine and suicide rates: suicide and the economy. PMID- 17132057 TI - Air travel and the spread of influenza: important caveats. PMID- 17132059 TI - City initiative: baby steps to a better future. PMID- 17132060 TI - Conjunctival FOXP3 in trachoma: T cells not specified. PMID- 17132061 TI - Health development versus medical relief: the illusion of aid. PMID- 17132062 TI - Intellectual property and access to ART: unwise choice of terminology. PMID- 17132064 TI - Spinal delivery of p38: TNF-alpha inhibitors. PMID- 17132065 TI - Are we publishing "the right stuff"? PMID- 17132066 TI - Photophysics and photochemical studies of 1,4-dihydropyridine derivatives. AB - The absorption and fluorescence properties of nifedipine (NPDHP), felodipine (CPDHP) and a series of structurally related 1,4-dihydropyridines were studied in aqueous solution and organic solvents of different properties. The absorption and fluorescence spectra were found to depend on the chemical nature of the substituents at the position 4 of the 1,4-dihydropyridine ring (DHP) and on solvent properties. In aqueous solution, the fluorescence spectra of 4-phenyl substituted compounds are blue-shifted with respect to the alkyl substituted compounds. The more fluorescent compound is CPDHP. Nifedipine is not fluorescent. All compounds, with the exception of CPDHP, present monoexponential fluorescence decay with very short lifetime (0.2-0.4 ns). CPDHP showed a biexponential emission decay with a long-lived component of 1.7 ns; this behavior is explained in terms of different conformers because of the hindered rotation of the phenyl group by the ortho-substitution. Analysis of the solvent effect on the maximum of the absorption spectrum by using the linear solvent-energy relation solvato chromic equation indicates the redshifts are influenced by the polarizability, hydrogen bonding ability and the hydrogen bond acceptance of the solvent. Whereas, the fluorescence characteristics (spectra, quantum yields and lifetimes) are sensitive to the polarizabilty and hydrogen bond ability of the solvents. Photo-decomposition of nifedipine is dependent on the solvent properties. Faster decomposition rates were obtained in nonprotic solvents. The 4-carboxylic derivative goes to decarboxylation. Under similar conditions, the other DHP compounds did not show appreciable photodecomposition. PMID- 17132067 TI - Sub-5-fs real-time spectroscopy of transition states in bacteriorhodopsin during retinal isomerization. AB - By using a sub-5-fs visible laser pulse, we have made the first observation of the vibrational spectra of the transition state during trans-cis isomerization in the retinal chromophore of bacteriorhodopsin (bR(S68). No instant isomerization of the retinal occurs in spite of electron promotion from the bonding pi-orbital to the anti-bonding pi*-orbital. The difference between the in-plane and out-of plane vibrational frequencies (about 1150-1250 and 900-1000 cm(-1), respectively) is reduced during the first time period. The vibrational spectra after this period became very broad and weak and are ascribed to a "silent state." The silent state lasts for 700-900 fs until the chromophore isomerizes to the cis-C13 = C14 conformation. The frequency of the C = C stretching mode was modulated by the torsion mode of the C13 = C14 double bond with a period of 200 fs. The modulation was clearly observed for four to five periods. Using the empirical equation for the relation between bond length and stretching frequency, we determined the transitional C = C bond length with about 0.01 angstroms accuracy during the torsion motion around the double bond with 1-fs time resolution. PMID- 17132068 TI - Structure and medium effects on the photochemical behavior of nonfluorinated quinolone antibiotics. AB - The photophysical behavior of the quinolone antibiotics, oxolinic (OX), cinoxacin (CNX) and pipemidic (PM) acids was studied as a function of pH and solvent properties. The ground state of these compounds exhibits different protonated forms, which also exist in the first excited states. Theoretical calculations of the Fukui indexes allowed to assigning the different protonation equilibria. The pK values indicate that the acidity of the 3-carboxylic and 4-carbonyl groups increases with the N-atom at position 2 in CNX. It has been found that fluorescence properties are strongly affected by pH, the more fluorescent species is that with protonated carboxylic acid, protonated species at the carbonyl group and the totally deprotonated form present very low fluorescence. The fluorescence behavior also depends on the chemical structure of the quinolone and on the solvent properties. The analysis of the solvent effect on the maximum and the width of the fluorescence band of OX, using the linear solvent-energy relation solvatochromic equation, indicates that the polarizability and hydrogen bond donor ability are the parameters that condition the spectral changes. The hydrogen bond acceptor ability of the solvents also contributes to the spectral shifts of CNX. The compound bearing the piperazinyl group at the position 7, PM only is fluorescent in high protic solvents. These results are discussed in terms of the competition between the intra- and intermolecular hydrogen bonds. The irradiation of OX, CNX and PM using 300 nm UV light led to a very low photodecomposition rate. Under the same conditions the nalidixic acid (NA), a structurally related quinolone, photodecomposes two orders of magnitude faster. PMID- 17132069 TI - OPC-67683, a nitro-dihydro-imidazooxazole derivative with promising action against tuberculosis in vitro and in mice. AB - BACKGROUND: Tuberculosis (TB) is still a leading cause of death worldwide. Almost a third of the world's population is infected with TB bacilli, and each year approximately 8 million people develop active TB and 2 million die as a result. Today's TB treatment, which dates back to the 1970s, is long and burdensome, requiring at least 6 mo of multidrug chemotherapy. The situation is further compounded by the emergence of multidrug-resistant TB (MDR-TB) and by the infection's lethal synergy with HIV/AIDS. Global health and philanthropic organizations are now pleading for new drug interventions that can address these unmet needs in TB treatment. METHODS AND FINDINGS: Here we report OPC-67683, a nitro-dihydro-imidazooxazole derivative that was screened to help combat the unmet needs in TB treatment. The compound is a mycolic acid biosynthesis inhibitor found to be free of mutagenicity and to possess highly potent activity against TB, including MDR-TB, as shown by its exceptionally low minimum inhibitory concentration (MIC) range of 0.006-0.024 microg/ml in vitro and highly effective therapeutic activity at low doses in vivo. Additionally, the results of the post-antibiotic effect of OPC-67683 on intracellular Mycobacterium tuberculosis showed the agent to be highly and dose-dependently active also against intracellular M. tuberculosis H37Rv after a 4-h pulsed exposure, and this activity at a concentration of 0.1 microg/ml was similar to that of the first line drug rifampicin (RFP) at a concentration of 3 microg/ml. The combination of OPC-67683 with RFP and pyrazinamide (PZA) exhibited a remarkably quicker eradication (by at least 2 mo) of viable TB bacilli in the lung in comparison with the standard regimen consisting of RFP, isoniazid (INH), ethambutol (EB), and PZA. Furthermore, OPC-67683 was not affected by nor did it affect the activity of liver microsome enzymes, suggesting the possibility for OPC-67683 to be used in combination with drugs, including anti-retrovirals, that induce or are metabolized by cytochrome P450 enzymes. CONCLUSIONS: We concluded that based on these properties OPC-67683 has the potential to be used as a TB drug to help combat the unmet needs in TB treatment. PMID- 17132070 TI - Does HIV cause cardiovascular disease? PMID- 17132071 TI - Health development versus medical relief: not a turf battle. PMID- 17132072 TI - 2,4-dichlorophenol degradation by an integrated process: photoelectrocatalytic oxidation and E-Fenton oxidation. AB - A new reactor system was designed for an integrated process involving photoelectrocatalytic oxidation (PECO) and electro-Fenton (E-Fenton) oxidation. Its efficiency was evaluated in terms of 2,4-dichlorophenol (2,4-DCP) degradation in aqueous solution. In this process, a TiO2 electrode and an iron (Fe) electrode were used as anodes in parallel, while graphite felt (GF) was used as a cathode. When an electrical current is applied between the anodes and the cathode, the iron anode can release Fe2+ and the GF cathode can generate H2O2 continuously in the reaction solution. Under UV-A illumination, while a H2O2-assisted PECO reaction occurs on the surface of the TiO2 photo anode, an E-Fenton reaction takes place in the solution. The experimental results demonstrated that 2,4-DCP degradation in aqueous solution was greatly enhanced because of the interaction between the two types of reactions. Moreover, the effect of pH as an important factor was investigated. It was found that the combined reaction becomes less pH sensitive than the typical E-Fenton reaction and may be suitable for application in a wide pH range. PMID- 17132073 TI - Animals roll around the clock: the rotation invariance of ultrarapid visual processing. AB - The processing required to categorize faces and animals is not only rapid but also remarkably resistant to inversion. It has been suggested that this sort of categorization performance could be achieved using the global distribution of orientations within the image, which interestingly is unchanged by inversion. Here, we presented subjects with two natural scenes at 16 different orientations that were simultaneously flashed in the left and right hemifield and we asked them to make a saccade to the side containing an animal. We report that human performance is surprisingly rotation invariant as reaction times were similar and accuracy remarkably stable across orientations. The results imply that this form of rapid object detection could not depend on the global distribution of orientations within the image. One alternative is that subjects are instead using local combinations of features that are diagnostic for the presence of an animal. PMID- 17132074 TI - Reversed phi revisited. AB - Two briefly flashed lines shown in rapid succession evoke a sensation of motion in human observers. This is examined quantitatively such that line separation, temporal offset, and contrast polarity are varied. Line pairs are presented on a gray background and are either both bright or both dark (equal contrast polarity) or one line is bright and the other is dark (opposite contrast polarity). Observers are instructed to indicate the perceived direction of motion. With foveal viewing, perceived direction is veridical for line pairs with equal contrast polarity but is reversed for line pairs with opposite contrast polarity, with spatial separations between 0 and 12 arcmin, and with temporal offsets between 8 and 33 ms. When separations, temporal offsets, or both are further increased, perception reverts to veridical. When lines of opposite contrast polarity are presented in the parafovea, reversal of perceived direction is also observed. The psychophysical results correlate well with those of recent intracellular recordings from directionally selective simple cells in the primary visual cortex of anesthetized cats (N. J. Priebe & D. Ferster, 2005). PMID- 17132075 TI - Forty-four years of studying light adaptation using the probed-sinewave paradigm. AB - Here we examine results from 44 years of probed-sinewave experiments investigating the dynamics of light adaptation. We also briefly examine four models that have been tested against the results. In these experiments, detection threshold is measured for a test stimulus superimposed at various times (phases) on a sinusoidally flickering homogeneous background. The results can be plotted as probe-threshold versus phase curves. Overall, the curves from different laboratories are remarkably similar given the substantial differences in experimental parameters. However, at medium frequencies of background flicker, there are some differences between the majority of the studies and a minority of two. An examination of the full set of results suggests that the differences are not as significant as they first appear and that the experimental condition leading to the differences is the use of long wavelength light in the two minority studies. Of the four models that have been tested, two fail to predict important features of the results, another is critically dependent on a mechanism unlikely to exist in the appropriate physiology, and the last seems quite promising. PMID- 17132076 TI - Kin recognition and the perceived facial similarity of children. AB - We examine the connection between a hypothetical kin recognition signal available in visual perception and the perceived facial similarity of children. One group of observers rated the facial similarity of pairs of children portrayed in photographs. Half of the pairs were siblings but the observers were not told this. A second group classified the pairs as siblings or nonsiblings. An optimal Bayesian classifier, given the similarity ratings of the first group, was as accurate in judging siblings as the second group. Mean rated similarity was also an accurate linear predictor (R2 = .96) of the log-odds that the rated pair portrayed were, in fact, siblings. Surprisingly, mean rated similarity did not vary with the age difference or gender difference of the pairs, both of which were counterbalanced across the stimuli. We conclude that the perceived facial similarity of children is little more than a graded kin recognition signal and that this kin recognition signal is effectively an estimate of the probability that two children are close genetic relatives. PMID- 17132077 TI - Crystalline lens radii of curvature from Purkinje and Scheimpflug imaging. AB - We present a comparison between measurements of the radius of the anterior and posterior lens surface, which was performed using corrected Scheimpflug imaging and Purkinje imaging in the same group of participants (46 for the anterior lens, and 34 for the posterior lens). Comparisons were also made as a function of accommodation (0 to 7 D) in a subset of 11 eyes. Data were captured and processed using laboratory prototypes and custom processing algorithms [for optical and geometrical distortion correction in the Scheimpflug system and using either equivalent mirror (EM) or merit function (MF) methods for Purkinje]. We found statistically significant differences in 4 of 46 eyes for the anterior lens radius, and 10 of 34 eyes for the posterior radius (using the MF and individual biometric data to process the Purkinje images). For the anterior lens, the agreement increases using individual biometry as opposed to biometric data from a model eye. For the posterior lens, the agreement increases using the MF as opposed to the EM method. For the changes during accommodation, no significant difference between the two techniques was found. In conclusion, the results of the cross-validation using the Scheimpflug and Purkinje imaging technique show that both techniques provide comparable lens radii and similar changes with accommodation. Purkinje tends to overestimate posterior lens radius, whereas pupil size limits the acquisition of posterior lens data with the Scheimpflug camera. Computer simulations using the Scheimpflug data as input show that the consistent slight overestimation of the posterior lens radius using Purkinje imaging can be partly attributed to the asphericity of the lens surface. PMID- 17132078 TI - Depth of interocular suppression associated with continuous flash suppression, flash suppression, and binocular rivalry. AB - When conflicting images are presented to the corresponding regions of the two eyes, only one image may be consciously perceived. In binocular rivalry (BR), two images alternate in phenomenal visibility; even a salient image is eventually suppressed by an image of low saliency. Recently, N. Tsuchiya and C. Koch (2005) reported a technique called continuous flash suppression (CFS), extending the suppression duration more than 10-fold. Here, we investigated the depth of this prolonged form of interocular suppression as well as conventional BR and flash suppression (FS) using a probe detection task. Compared to monocular viewing condition, CFS elevated detection thresholds more than 20-fold, whereas BR did so by 3-fold. In subsequent experiments, we dissected CFS into several components. By manipulating the number and timing of flashes with respect to the probe, we found that the stronger suppression in CFS is not due to summation between BR and FS but is caused by the summation of the suppression due to multiple flashes. Our results support the view that CFS is not a stronger version of BR but is due to the accumulated suppressive effects of multiple flashes. PMID- 17132079 TI - The flight path of the phoenix--the visible trace of invisible elements in human vision. AB - How features are attributed to objects is one of the most puzzling issues in the neurosciences. A deeply entrenched view is that features are perceived at the locations where they are presented. Here, we show that features in motion displays can be systematically attributed from one location to another although the elements possessing the features are invisible. Furthermore, features can be integrated across locations. Feature mislocalizations are usually treated as errors and limits of the visual system. On the contrary, we show that the nonretinotopic feature attributions, reported herein, follow rules of grouping precisely suggesting that they reflect a fundamental computational strategy and not errors of visual processing. PMID- 17132080 TI - Induced movement: the flying bluebottle illusion. AB - Two small objects (flies) followed identical circular orbits. However, a large background that circled around behind them in different phases made one orbit look twice as large as the other (size illusion) or made the circles look like very thin horizontal or vertical ellipses with aspect ratios of 7.5:1 or more (shape illusion). The nature of the perceptual distortion depended upon the relative phase between the movements of the background and those of the flies. Brief snatches of the moving background that added up to a circular motion were also effective. PMID- 17132081 TI - Induced visual fading of complex images. AB - Visual stimuli fade from awareness under retinal stabilization or careful fixation, a phenomenon documented by Troxler more than 200 years ago. Research on visual fading during normal visual fixation typically has been restricted to discrete, simple, low-contrast shapes presented peripherally against a uniform or textured background. In four experiments, we document a striking new visual fading effect in which entire photographs of scenes fade to a uniform luminance and hue during normal visual fixation. Critically, this "scene fading" can be induced almost instantaneously by some types of visual transients but not by others. These induced fading effects are sufficiently robust that they can be experienced by most observers in a single trial. Taken as a whole, the effects are inconsistent with simple contrast adaptation, gradual Troxler fading, or transient-induced fading. They are, however, consistent with the idea that small contrast decrements can induce fading of entire scenes. The methods provide a robust tool for the exploration of visual fading, and the results could have important implications for the role of filling-in and neural adaptation in our visual awareness of natural scenes and other complex stimuli. PMID- 17132082 TI - Color constancy and hue scaling. AB - In this study, we used a hue scaling technique to examine human color constancy performance in simulated three-dimensional scenes. These scenes contained objects of various shapes and materials and a matte test patch at the center of the scene. Hue scaling settings were made for test patches under five different illuminations. Results show that subjects had nearly stable hue scalings for a given test surface across different illuminants. In a control experiment, only the test surfaces that belonged to one illumination condition were presented, blocked in front of a black background. Surprisingly, the hue scalings of the subjects in the blocked control experiment were not simply determined by the color codes of the test surface. Rather, they depended on the sequence of previously presented test stimuli. In contrast, subjects' hue scalings in a second control experiment (with order of presentations randomized) were completely determined by the color codes of the test surface. Our results show that hue scaling is a useful technique to investigate color constancy in a more phenomenological sense. Furthermore, the results from the blocked control experiment underline the important role of slow chromatic adaptation for color constancy. PMID- 17132083 TI - Spatial summation of face information. AB - Do all parts of the face contribute equally to face detection or are some parts more detectable than others? To evaluate this issue, we studied detection of the presence of normalized frontal-face images within aperture windows of varying extent. We performed a face summation study using two-alternative forced-choice psychophysics. The face stimuli were scaled to equal eye-to-chin distance, centered on the bridge of the nose, and windowed by fourth-power Gaussian envelopes of various sizes. The faces were intermixed with control stimuli consisting inverted faces to test for configuration effects, split-half inverted faces to perturb the symmetry, and phase-scrambled versions of the faces with equal Fourier energy. Face detectability improved rapidly at first, then at a progressively shallower rate for larger window sizes, in a similar fashion for the three face-based stimulus types. The spectrally equated noise stimuli were less detectable than the face stimuli for all except the smallest apertures. The results were fit with a model incorporating global face-specific and local nonspecific spatial integration mechanisms. Detection of the noise images was consistent with local detection mechanisms accessed through a wide-field attention mechanism. The data for face detection implied detection mechanisms that integrated linearly up to some small size, integrated more slowly up to an intermediate size, and failed to gain any improvement for information beyond some larger size. This performance supports the concept of a specialized face configuration mechanism operating at detection threshold, similar in extent among the observers. PMID- 17132084 TI - Edge integration and the perception of brightness and darkness. AB - How do induced brightness and darkness signals from local and remote surfaces interact to determine the final achromatic color percept of a target surface? An emerging theory of achromatic color perception posits that brightness and darkness percepts are computed by weighting and summing the induction signals generated at edges in a scene. This theory also characterizes how neighboring edges interact to modulate the gain of brightness and darkness signals induced from one another. Here we assess evidence for this edge integration theory by means of computational modeling and a psychophysical experiment. We quantitatively show how local and remote edge induction signals in disk-ring displays give rise to either contrast or assimilation effects. Spatial integration of same-polarity edge signals supports a contrast effect, whereas integration of opposite-polarity signals supports an assimilation effect, particularly when the remote induction signal is much stronger than the local induction signal. The results confirm a key prediction of edge integration theory, namely, that strong assimilation effects can lead subjects to ignore the polarity of local edge information when setting achromatic color matches. The conditions necessary for strong assimilation effects are also associated with greater difficulty in setting matches, suggesting that caution is required when interpreting matching data in terms of gain control. We describe several avenues for further study of contrast, assimilation, and gain control. PMID- 17132085 TI - Structure of interphase chromosomes in the nuclei of Drosophila cells. AB - Fluorescent images of interphase chromatin structures and chromosome structures isolated from reversibly permeable Drosophila cells were analyzed. Decondensed chromatin in early S phase (2.0-2.5 C-value) consisted of a veil-like fibrillary network. Fibrillar chromatin formed rodlets later in the early S phase (2.5-2.75 C). Drosophila chromosomes contain several smaller subunits called rodlets. Fibrillar chromatin turned to chromatin ribbon and the early mid-S-phase globular chromosomes (2.75-3.0 C), then to opened fibrous globular forms later in the mid S-phase (3.0-3.25 C), to late-S-phase supercoiled ribbons (3.25-3.5 C), end-S phase elongated prechromosomes (3.5-3.75 C), bent and linear chromosomes (3.75 4.0 C). Early-S phase chromatin fibrils in the nuclei of Drosophila cells are thinner than the veil-like structures in mammalian cells. The connectivity of chromosomes shows linear arrangement (3, 1, 2, 4), with larger chromosomes (1 and 2) inside and smaller chromosomes (3, 4) at the two ends in the chromosomal chain. PMID- 17132086 TI - Characterization of the structures involved in localization of the SUN proteins to the nuclear envelope and the centrosome. AB - The nuclear envelope forms a selective barrier that separates the cytoplasm from the nucleus. During mitosis the nuclear envelope breaks down so that the microtubule network can form contacts with the kinetochore and guide chromosome segregation. Previous studies have suggested a model in which the centrosome and the microtubule network may play a role in nuclear envelope breakdown through as yet unidentified interactions with proteins localized to the nuclear envelope. In the current study we characterized a nuclear envelope protein SUN2 and identified a substructure involved in its localization to the nuclear envelope. We found that a structurally related protein, SUN1, may be localized to the nuclear envelope through a different mechanism. Furthermore, the SUN2 protein can form different assemblies, including homodimers and heterodimers with SUN1. Finally, we provide evidence indicating that SUN1 and SUN2 may form a physical interaction between the nuclear envelope and the centrosome. PMID- 17132087 TI - Autoantibodies closely relate to the elevation level of in vivo hydrogen peroxide and tissue damage in systemic lupus erythematosus. AB - Recent studies have shown that antibodies efficiently catalyze the conversion of molecular singlet oxygen (1O2) plus water to hydrogen peroxide (H2O2). H2O2 is toxic to cells and is a cause of further free radical generation that are implicated in the pathogenesis of a variety of diseases. Systemic lupus erythematosus (SLE) is one of the most serious autoimmune diseases which are characterized by the production of various autoantibodies and subsequent tissue damage. However, the correlation of autoantibodies, H2O2, and tissue lesion in SLE has not been yet investigated. To address this issue, in the present study, we induced autoantibodies and kidney tissue damage by using SLE animal model as described previously. We detected the level of H2O2 in SLE mice and found the increase of in vivo H2O2 was accompanied and closely correlated with the production of anti-dsDNA and antihistone antibodies. Importantly, there was onefold increase of H2O2 in the mice kidneys with apparent glomerulonephritis and IgG deposits. These results suggest that the induced autoantibodies possess catalytic activity. The produced autoantibodies lead to the production and elevation of H2O2, which results in subsequent renal damage and the pathogenesis of SLE. Our findings provide an insight into the understanding of SLE mechanism and provide a potential approach for therapeutic intervention of SLE. PMID- 17132089 TI - Human retinal and brain cell lines: A model of HCMV retinitis and encephalitis. AB - Although HIV is accepted as the etiologic agent in AIDS, other factors have been implicated in accelerating the disease. Human cytomegalovirus (HCMV) in particular has been implicated as a cofactor in the progression from AIDS-related complex (ARC) to AIDS. HCMV infection of the central nervous system (CNS) (brain, retina) has been reported in at least 50% of AIDS patients, and has been implicated in producing encephalitis and sight-threatening retinitis. HCMV exhibits strict species specificity and animal models for human HCMV are conspicuous by their absence. We have developed a human brain cell line (mixed glial/neuronal) and a multipotential human retinal precursor cell line (neuronal in nature). We have tested the suitability of these cell lines as models for the study of HCMV infectibility. In this study, we report that these cell lines are optimal for the study of HCMV infectibility and pathogenesis in tissues of neural origin and appropriate to study HIV-HCMV interaction. Immortalized human brain and retinal cell lines were infected with a laboratory strain of HCMV (AD 169, Towne) at a multiplicity of infection moi (1-5) and viral infectibility and cell specificity monitored by: (a) phenotypic analysis (multinucleate cells, syncytium formation, etc.), (b) antigen expression (IE, E, late) by immunohistochemistry, Western blot analysis, (c) presence of viral particles by TEM, and (d) expression of indicator plasmids (HIV-LTR-CAT). We report that both human retinal and brain cell lines are permissive for HCMV infectibility. Cell specificity was not seen; both cells expressing glial/neuronal cell markers were positive for the presence of HCMV early/late antigens. Formation of multinucleate giant cells with nuclear inclusion bodies and syncytia were seen. Productive viral infection was confirmed by the ability of cell-free supernatant from the third passage of infected cells to produce pathogenicity and express viral particles, when added to fresh cultures. Using indicator plasmids, HIV-LTR, and CAT, we have shown that HIV and HCMV interact at the cellular level. We have also shown that HIV production in retinal and brain cell lines transfected with cloned HIV was enhanced by HCMV-IE genes. We did not see any differences in HCMV. AD 169, Towne isolate, and data from both strains is presented in this paper. This model could prove extremely useful for the study of cell specificity/cellular and molecular interaction between HIV/HCMV and to test antiviral therapies. PMID- 17132088 TI - Abeta-immunotherapy for Alzheimer's disease using mannan-amyloid-Beta peptide immunoconjugates. AB - In Alzheimer's disease (AD) the accumulation of pathological forms of the beta amyloid (Abeta) peptide are believed to be causal factors in the neurodegeneration that results in the loss of cognitive function in patients. Anti-Abeta antibodies have been shown to reduce Abeta levels in transgenic mouse models of AD and in AN-1792 clinical trial on AD patients; however, the clinical trial was halted when some patients developed meningoencephalitis. Theories on the cause of the adverse events include proinflammatory "primed patients," a Th1 inducing adjuvant, and Abeta autoreactive T cells. New immunotherapy approaches are being developed to eliminate these putative risk factors. Mannan, which is recognized by pattern recognition receptors of the innate immune system, can be utilized as a molecular adjuvant to promote a Th2-mediated immune response to conjugated B cell epitopes. The N-terminus of Abeta was conjugated to mannan, and used to immunize mice with low concentrations of immunoconjugate, without a conventional adjuvant. Mannan induced a significant and highly polarized toward Th2 phenotype anti-Abeta antibody response not only in BALB/c, but also in B6SJL F1 mice. New preclinical trials in AD mouse models may help to develop novel immunogen-adjuvant configurations with the potential to avoid the adverse immune response that occurred in the first clinical trial. PMID- 17132090 TI - Characterization of frequently deleted 6q locus in prostate cancer. AB - The long arm of chromosome 6 is frequently deleted in diverse human neoplasms. Our previous study showed a minimum deletion region between markers D6S1056 and D6S300 on chromosome 6q in primary prostate cancer (CaP). In this study, we further refined a 200-kb minimal region of deletion (6qTSG1) centered around D6S1013 marker. The 6qTSG1 transcripts contained complex multiple splicing variants with low or absent expression in CaP cells. None of the transcripts identified contained open reading frames that code for a protein in the NCBI database. The expression of 6qTSG transcripts revealed interesting hormonal regulation relevant to CaP biology. Expression of 6q TSG transcript was induced in LNCaP cells that were cultured in charcoal-stripped serum medium suggesting an upregulation of 6qTSG transcript by androgen ablation and cell growth inhibition/apoptosis. Induction of 6qTSG1 expression in response to androgen ablation was abrogated in androgen-independent derivatives of LNCaP cells. In summary, we have defined a candidate CaP suppressor locus on chromosome 6q16.1, and deletions of this locus are frequently associated with prostate tumorigenesis. In the light of emerging role of noncoding RNAs in cancer biology including CaP, future investigations of 6qTSG11 locus is warranted. PMID- 17132091 TI - Expression profiling of Yersinia pestis during mouse pulmonary infection. AB - Yersinia pestis, the causative agent of plague, can be transmitted by infected flea bite or inhaled aerosol. Both routes of infection have a high mortality rate, and pneumonic infections of Y. pestis represent a significant concern as a tool of bioterrorism. Understanding the transcriptional program of this pathogen during pulmonary infection should be valuable in understanding plague pathogenesis, as well as potentially offering insights into new vaccines and therapeutics. Toward this goal we developed a long oligonucleotide microarray to the plague bacillus and evaluated the expression profiles of Y. pestis in vitro and in the mouse pulmonary infection model in vivo. The in vitro analysis compared expression patterns at 27 versus 37 degrees C, as a surrogate of the transition from the flea to the mammalian host. The in vivo analysis used intranasal challenge to the mouse lung. By amplifying the Y. pestis RNA from individual mouse lungs we were able to map the transcriptional profile of plague at postinfection days 1 to 3. Our data present a very different transcriptional profile between in vivo and in vitro expression, suggesting Y. pestis responds to a variety of host signals during infection. Of note was the number of genes found in genomic regions with altered %GC content that are upregulated within the mouse lung environment. These data suggest these regions may provide particularly promising targets for both vaccines and therapeutics. PMID- 17132092 TI - Partitioning of aminoacyl-tRNA synthetases in two classes could have been encoded in a strand-symmetric RNA world. AB - The "chicken-or-egg" dilemma dictates that archaic tRNAs be aminoacylated by ribozymic aminoacyl-tRNA synthetases, rAARSs, with protein synthetases (pAARSs) emerging later and, strikingly in two versions. However, the distribution of these two versions among the codons also suggests their involvement in development of the genetic code. Here we propose a solution to this controversy, which relies on a primordial complementarity hypothesis that in a strand symmetric RNA world both complementary replicas of many genes could encode the first proteins. Accordingly, if one rearranges the code table in a manner that puts complementary codons directly against each other, an almost perfect mirror symmetry in tRNA aminoacylation by the two groups of synthetases is revealed. Specifically, the pairs of complementary anticodons from the same pAARS class tend to contain RR and YY dinucleotides at first and second versus third and second positions, whereas in pairs of pAARSs from the different classes these positions are occupied by YR and RY, including CG, GC, UA, and AU palindromes. The latter are indistinguishable in complementary anticodons, thus leading to erroneous aminoacylation (note that there is no such problem for RR- and YY containing complementary anticodons). This can be averted by "spreading out" tRNA recognition by two rAARSs away from the anticodons in the opposite directions, giving two complementary rAARSs. The principle of evolutionary continuity suggests that their protein successors also arose on complementary strands. Our analyses support this hypothesis. PMID- 17132093 TI - Amplification of sense-stranded prokaryotic RNA. AB - Microarray expression analysis has proven to be a valuable methodology. In eukaryotic systems where RNA is limiting, established protocols for amplification of mRNA, which rely on the poly(A) tails, are well established. In contrast, the difficulty in amplifying prokaryotic mRNA has limited the application of microarrays to microbiology. Here we present a method for the Linear Amplification of Prokaryotic Transcripts (LAPT) that is efficient and unbiased. The overhang tailing activity of Moloney murine leukemia virus reverse transcriptase is used to add the T7 promoter to cDNAs during reverse transcription. The promoter addition is uncoupled from the initial priming event allowing the promoter to be attached to the 5' end of the RNA transcript. This enables the amplification of sense-stranded RNA that is representative of the complexity and distribution of the original transcript pool. In microarray assays amplified prokaryotic RNA (10 ng total RNA starting material) showed good Spearman correlations to an unamplified control sample. Using genome-directed primers to bias addition of a T7-promoter to bacterial transcripts allowed amplification of prokaryotic transcripts in the presence of mammalian RNA (at a eukaryotic/prokaryotic RNA ratio of 500 to 1). This technology should facilitate the study of prokaryotic transcriptomes in situations, such as in vivo studies or mixed microbial populations, where the prokaryotic RNA amount is limited and/or the nontarget/target RNA ratios is high. PMID- 17132094 TI - Quantification of fetal and total circulatory DNA in maternal plasma samples before and after size fractionation by agarose gel electrophoresis. AB - Fetal extracellular DNA is mainly derived from apoptotic bodies of trophoblast. Recent studies have shown size differences between fetal and maternal extracellular DNA. We have examined the quantification of fetal (SRY gene) and total (GLO gene) extracellular DNA in maternal plasma in different fractions (100 300, 300-500, 500-700, 700-900, and >900 bp) after size fractionation by agarose gel electrophoresis. DNA was extracted from maternal plasma samples from 11 pregnant women carrying male foetuses at the 16th week of gestation. Fetal circulatory DNA was mainly detected in the 100-300 bp fraction with the median concentration being 14.4 GE/ml. A lower median amount of 4.9 GE/ml was also found in the 300-500 bp fraction. Circulatory DNA extracted from the 100-300 bp fraction contained 4.2 times enriched fetal DNA when compared with unseparated DNA sample. Fetal DNA within the 300-500 bp fraction was 2.5 times enriched. Circulatory fetal DNA is predominantly present in a fraction with molecular size <500 bp, which can be used for the detection of paternally inherited alleles. However, the usage of size-separated DNA is not suitable for routine clinical applications because of risk of contamination. PMID- 17132095 TI - Condensation of interphase chromosomes in nuclei of synchronized CHO cells. AB - The escape of individual interphase chromosomes from nuclei of reversibly permeabilized Chinese hamster ovary (CHO) cells was utilized for the visualization of condensing interphase chromosomes in a cell cycle-dependent manner in synchronized cells. Major interphase chromosomal forms include: (a) mid S-phase globular chromosomes at 3.0 C-value, (b) late mid-S-phase fibrous hemicircular forms (3.3 C), (c) late-S-phase supercoiled ribbons (3.7 C), and (d) end-S-phase elongated, bent prechromosomal structures (4.0 C). PMID- 17132096 TI - In vitro expression and monoclonal antibody of RNA-dependent RNA polymerase for infectious bursal disease virus. AB - VP1, the RNA-dependent RNA polymerase of infectious bursal disease virus (IBDV), has been suggested to play an essential role in the replication and translation of viral RNAs. In this study, we first expressed the complete VP1 protein gene in Escherichia coli (E. coli), and then the produced polyclonal antibody and four monoclonal antibodies (mAbs) to recombinant VP1 protein (rVP1) were shown to bind the IBDV particles in chicken embryo fibroblast and Vero cells. The epitopic analysis showed that mAbs 1D4 and 3C7 recognized respectively two distinct antigenic epitopes on the rVP1 protein, but two pair of mAbs 1A2/2A12 and 1E1/1H3 potentially recognized another two topologically related epitopes. Immunocytochemical stainings showed that VP1 protein formed irregularly shaped particles in the cytoplasm of the IBDV-infected cells. These results demonstrated that the mAbs to rVP1 protein could bind the epitopes of IBDV particles, indicating that the rVP1 protein expressed in E. coli was suitable for producing the mAb to VP1 protein of IBDV, and that the cytoplasm could be the crucial site for viral genome replication of IBDV. PMID- 17132097 TI - Janus-faced role of endothelial NO synthase in vascular disease: uncoupling of oxygen reduction from NO synthesis and its pharmacological reversal. AB - Endothelial NO synthase (eNOS) is the predominant enzyme responsible for vascular NO synthesis. A functional eNOS transfers electrons from NADPH to its heme center, where L-arginine is oxidized to L-citrulline and NO. Common conditions predisposing to atherosclerosis, such as hypertension, hypercholesterolemia, diabetes mellitus and smoking, are associated with enhanced production of reactive oxygen species (ROS) and reduced amounts of bioactive NO in the vessel wall. NADPH oxidases represent major sources of ROS in cardiovascular pathophysiology. NADPH oxidase-derived superoxide avidly interacts with eNOS derived NO to form peroxynitrite (ONOO(-)), which oxidizes the essential NOS cofactor (6R-)5,6,7,8-tetrahydrobiopterin (BH(4)). As a consequence, oxygen reduction uncouples from NO synthesis, thereby rendering NOS to a superoxide producing pro-atherosclerotic enzyme. Supplementation with BH(4) corrects eNOS dysfunction in several animal models and in patients. Administration of high local doses of the antioxidant L-ascorbic acid (vitamin C) improves endothelial function, whereas large-scale clinical trials do not support a strong role for oral vitamin C and/or E in reducing cardiovascular disease. Statins, angiotensin converting enzyme inhibitors and AT1 receptor blockers have the potential of reducing vascular oxidative stress. Finally, novel approaches are being tested to block pathways leading to oxidative stress (e.g. protein kinase C) or to upregulate antioxidant enzymes. PMID- 17132098 TI - Leucine aminopeptidases: diversity in structure and function. AB - Leucine aminopeptidases (LAPs) are metallopeptidases that cleave N-terminal residues from proteins and peptides. While hydrolyzing Leu substrates, LAPs often have a broader specificity. LAPs are members of the M1 or M17 peptidase families, and therefore the LAP nomenclature is complex. LAPs are often viewed as cell maintenance enzymes with critical roles in turnover of peptides. In mammals, the M17 and M1 enzymes with LAP activity contribute to processing peptides for MHC I antigen presentation, processing of bioactive peptides (oxytocin, vasopressin, enkephalins), and vesicle trafficking to the plasma membrane. In microbes, the M17 LAPs have a role in proteolysis and have also acquired the ability to bind DNA. This property enables LAPs to serve as transcriptional repressors to control pyrimidine, alginate and cholera toxin biosynthesis, as well as mediate site specific recombination events in plasmids and phages. In plants the roles of the M17 LAPs and the peptidases related to M1 LAPs are being elucidated. Roles in defense, membrane transport of auxin receptors, and meiosis have been implicated. PMID- 17132099 TI - Endogenous anti-inflammatory substances, inter-alpha-inhibitor and bikunin. AB - There have been new developments in the elucidation of the biological functions of the inter-alpha-inhibitor (IalphaI) family. The anti-proteolytic activity of the IalphaI family originates from bikunin (also known as urinary trypsin inhibitor). Growing evidence indicates that bikunin is not just an anti proteolytic agent, but can also be considered an anti-inflammatory agent that suppresses lipopolysaccharide (LPS)-induced cytokine synthesis. Bikunin functions to inhibit calcium influx and extracellular signal-regulated kinase (ERK) signaling via LPS receptors and/or as yet unidentified bikunin signaling receptors. By signaling via the LPS receptor, LPS increases calcium influx and yields phosphorylated ERK, which activates multiple transcription factors, such as nuclear factor kappaB (NF-kappaB) or early growth response-1 (Egr-1), which in turn promote cytokine expression. Deficits in the signaling cascades caused by free or cell-bound bikunin are predicted to down-regulate cytokine expression, render macrophages/neutrophils more inactive, and impair inflammatory processes. This brief review largely focuses on our current understanding of the apparent functions of bikunin, its ligands, the effector molecules with which it interacts, and its regulation. PMID- 17132100 TI - Mitochondrial morphology and distribution in mammalian cells. AB - It is now appreciated that mitochondria form tubular networks that adapt to the requirements of the cell by undergoing changes in their shape through fission and fusion. Proper mitochondrial distribution also appears to be required for ATP delivery and calcium regulation, and, in some cases, for cell development. While we now realise the great importance of mitochondria for the cell, we are only beginning to work out how these organelles undergo the drastic morphological changes that are essential for cellular function. Of the few known components involved in shaping mitochondria, some have been found to be essential to life and their gene mutations are linked to neurological disorders, while others appear to be recruited in the activation of cell death pathways. Here we review our current understanding of the functions of the main players involved in mitochondrial fission, fusion and distribution in mammalian cells. PMID- 17132101 TI - Heterogeneity in the cysteine protease inhibitor clitocypin gene family. AB - Clitocypin from the basidiomycete Clitocybe nebularis is the first fungal protein cysteine protease inhibitor to be characterised in detail, yet no information on its molecular genetics is available. Owing to its unique characteristics, it was assigned as the only member of a new family of cysteine protease inhibitors in the MEROPS inhibitor classification. Here we describe the full-length sequence of the clitocypin gene. A BLAST search confirmed its lack of significant sequence similarity to any other gene. The gene is composed of four exons and three short introns and belongs to a small family of closely related genes with more than 90% identity. Sequence variability is evenly distributed in introns and exons and deduced amino acid substitutions are distributed throughout the protein sequence. Basidiocarps collected at two distant locations were examined and the level of heterogeneity found in one basidiocarp is similar to that between the two. Sequencing of the ribosomal DNA spacers from the two basidiocarps confirmed that the heterogeneity observed in the clitocypin gene is not due to evolutionary divergence of the two specimens caused by geographic separation. Clitocypin is expressed in different parts of the basidiocarp and in cultured mycelia in a manner suggesting regulation by developmental and/or environmental factors. PMID- 17132102 TI - Mutations in the inter-SH2 domain of the regulatory subunit of phosphoinositide 3 kinase: effects on catalytic subunit binding and holoenzyme function. AB - Class IA phosphoinositide 3-kinases (PI3Ks) represent a group of heterodimeric lipid kinases with important functions in cellular signal transduction. The regulatory p85 subunit constitutively binds to the catalytic p110 subunit and mediates the recruitment of the heterodimer to various membrane-localized proteins upon activation by a vast array of stimuli. The functional characterization of protein domains that mediate p85 function has been hampered by a lack of structural data. Therefore, we investigated a 35-aa region in the inter-SH2 domain of p85, reported to be necessary for binding of p110, by site directed mutagenesis and evaluated the importance of individual amino acids for PI3K heterodimer formation. This approach led to the identification of an 11-aa region required for p110 binding in vitro and mesoderm induction during early Xenopus development in vivo. Further analyses revealed two pairs of hydrophobic amino acids within this region, which are particularly important for high affinity intersubunit interaction. Thus, our data provide further insight into the molecular mechanisms of PI3K intersubunit interaction and led to the identification of new p85 mutant proteins with varying degrees of dominant negative effects that will be helpful for future PI3K-related research. PMID- 17132103 TI - Evaluation of Bacillus anthracis thymidine kinase as a potential target for the development of antibacterial nucleoside analogs. AB - Bacillus anthracis, which causes anthrax, has attracted attention because of its potential use as a biological weapon. The risk of multidrug resistance against B. anthracis increases the need for antibiotics with new molecular targets. Nucleoside analogs are well-known antiviral and anticancer prodrugs, and thymidine kinase catalyzes the rate-limiting step in the activation of pyrimidine nucleoside analogs used in chemotherapy. The thymidine kinase gene from B. anthracis Sterne strain (34F2) (Ba-TK) was cloned and expressed in E. coli, and the product was purified and characterized regarding its substrate specificity. Ba-TK phosphorylated pyrimidine nucleosides and all natural nucleoside triphosphates served as phosphate donors. Size exclusion chromatography indicated a dimeric form of Ba-TK, regardless of the presence of ATP. Thymidine was the most efficient substrate with a low K(m) value (0.6 microM) and a V(max) of 3.3 micromol dTMP mg(-1) min(-1), but deoxyuridine (K(m)=4.2 microM, V(max)=4.1 micromol dUMP mg(-1) min(-1)) was also a good substrate. Several pyrimidine analogs were also tested and analogs with 5-position modifications showed higher activities compared to analogs with 3'- and N3-position modifications. Deoxyuridine analogs were the most potent inhibitors of B. anthracis growth in vitro. These results may be used to guide future development of nucleoside analogs against B. anthracis. PMID- 17132104 TI - The human malaria parasite Plasmodium falciparum expresses an atypical N terminally extended pyrophosphokinase with specificity for thiamine. AB - Vitamin B(1) is an essential cofactor for key enzymes such as 2-oxoglutarate dehydrogenase and pyruvate dehydrogenase. Plants, bacteria and fungi, as well as Plasmodium falciparum, are capable of synthesising vitamin B(1)de novo, whereas mammals have to take up this cofactor from their diet. Thiamine, a B(1) vitamer, has to be pyrophosphorylated by thiamine pyrophosphokinase (TPK) to the active form. The human malaria parasite P. falciparum expresses an N-terminally extended pyrophosphokinase throughout the entire erythrocytic life cycle, which was analysed by Northern and Western blotting. The recombinant enzyme shows a specific activity of 27 nmol min(-1) mg(-1) protein and specificity for thiamine with a K(m) value of 73 microM, while thiamine monophosphate is not accepted. Mutational analysis of the N-terminal extension of the plasmodial TPK showed that it influences thiamine binding as well as metal dependence, which suggests N terminal participation in the conformation of the active site. Protein sequences of various plasmodial TPKs were analysed for their phylogeny, which classified the Plasmodium TPKs to a group distinct from the mammalian TPKs. To verify the location of the parasite TPK within the cell, immunofluorescence analyses were performed. Co-staining of PfTPK with a GFP marker visualised its cytosolic localisation. PMID- 17132105 TI - Fes1p acts as a nucleotide exchange factor for the ribosome-associated molecular chaperone Ssb1p. AB - The HspBP1 homolog Fes1p was recently identified as a nucleotide exchange factor (NEF) of Ssa1p, a canonical Hsp70 molecular chaperone in the cytosol of Saccharomyces cerevisiae. Besides the Ssa-type Hsp70s, the yeast cytosol contains three additional classes of Hsp70, termed Ssb, Sse and Ssz. Here, we show that Fes1p also functions as NEF for the ribosome-bound Ssb Hsp70s. Sequence analysis indicated that residues important for interaction with Fes1p are highly conserved in Ssa1p and Ssb1p, but not in Sse1p and Ssz1p. Indeed, Fes1p interacts with Ssa1p and Ssb1p with similar affinity, but does not form a complex with Sse1p. Functional analysis showed that Fes1p accelerates the release of the nucleotide analog MABA-ADP from Ssb1p by a factor of 35. In contrast to the interaction between mammalian HspBP1 and Hsp70, however, addition of ATP only moderately decreases the affinity of Fes1p for Ssb1p. Point mutations in Fes1p abolishing complex formation with Ssa1p also prevent the interaction with Ssb1p. The ATPase activity of Ssb1p is stimulated by the ribosome-associated complex of Zuotin and Ssz1p (RAC). Interestingly, Fes1p inhibits the stimulation of Ssb1p ATPase by RAC, suggesting a complex regulatory role of Fes1p in modulating the function of Ssb Hsp70s in co-translational protein folding. PMID- 17132106 TI - Detailed analysis of MIA protein by mutagenesis. AB - MIA (melanoma inhibitory activity) has been identified as a small protein secreted by malignant melanoma cells that interacts with extracellular matrix proteins including fibronectin. These findings suggest that MIA may play a role in tumor progression and the spread of malignant melanomas by mediating detachment of cells from extracellular matrix molecules. Here, we present a detailed study on functionally important MIA domains. Using site-directed mutagenesis, amino acids important for MIA structure and/or function were determined. Amino acids conserved in SH3 domains were shown to be important for structural integrity. In addition, amino acid residues necessary for MIA function were identified. Interestingly, not all of them are conserved with respect to other members of the MIA protein family. In summary, our results lead to a better understanding of MIA function. Regulating MIA functions in vivo may provide a novel therapeutic strategy for metastatic melanoma disease. PMID- 17132107 TI - The role of human tissue kallikreins 7 and 8 in intracranial malignancies. AB - Recent evidence suggests that many tissue kallikreins are implicated in carcinogenesis. Kallikrein 8 (KLK8) plays a role in the physiology of the central nervous system. Kallikrein 7 (KLK7) takes part in skin desquamation. Both show altered expression in ovarian and breast cancer. In this study, we examined the level of mRNA expression of the KLK7 and KLK8 genes in 73 intracranial tumors using qualitative RT-PCR. The results were correlated with clinical and histomorphological variables and patient outcome. The expression of both genes was also examined in the brain cancer cell lines U-251 MG, D54 and SH-SY5Y and the invasive capacity of glioblastoma cells U-251 MG overexpressing hK7 or hK8 was also investigated in an in vitro Matrigel assay. Follow-up analysis revealed that expression of KLK7 mRNA was associated with shorter overall survival (OS) compared to patients with no KLK7 expression, as determined by Cox proportional hazard regression analysis. Overexpression of hK7 protein by cultivated brain tumor cells significantly enhanced the invasive potential in the Matrigel invasion assay, in contrast to cells overexpressing hK8 protein. Our data suggest that hK7 protein overexpression is associated with a more aggressive phenotype in brain cancer cells. PMID- 17132108 TI - Prognostic significance of the expression of SR-A1, encoding a novel SR-related CTD-associated factor, in breast cancer. AB - SR-A1 is a human high-molecular-weight SR-related CTD-associated factor that links the machineries of transcription and mRNA splicing. In this study we examined the prognostic value of SR-A1 gene expression using a semi-quantitative RT-PCR method. High SR-A1 expression was observed in 31/81 (38.3%) breast cancer tissues and was found to be more frequent in patients with tumors of large size (p=0.027), as well as in lymph node-positive patients (p=0.035). Follow-up analysis revealed that low SR-A1 expression increases the probability of both overall and disease-free survival of patients. Our results suggest that SR-A1 may possibly be characterized as a new marker of unfavorable prognosis for breast cancer. PMID- 17132109 TI - Suppression of TNF-alpha production by S-adenosylmethionine in human mononuclear leukocytes is not mediated by polyamines. AB - Endotoxin-induced cytokine production is an important mechanism in the development of several types of liver damage. Methionine, some of its precursors and metabolites were reported to have protective effects against such injury. The aim of this study was to investigate whether methionine, its precursors or metabolites [phosphatidylcholine, choline, betaine, S-adenosylmethionine (SAM)] have a modulating effect on tumor necrosis factor alpha (TNF-alpha) production by endotoxin-stimulated human mononuclear leukocytes and whether SAM-dependent polyamines (spermidine, spermine) are mediators of SAM-induced inhibition of TNF alpha synthesis. Methionine and betaine had a moderate stimulatory effect on TNF alpha production, whereas phosphatidylcholine (ID(50) 5.4 mM), SAM (ID(50) 131 microM), spermidine (ID(50) 4.5 microM) and spermine (ID(50) 3.9 microM) had a predominantly inhibitory effect. Putrescine did not alter TNF-alpha release. Inhibitors of polyamine synthesis that blocked either putrescine (difluoromethylornithine) or spermine (CGP48664A) production did not affect TNF alpha synthesis. Endotoxin stimulation of leukocytes did not alter the intracellular levels of polyamines. In addition, supplementation with SAM did not change the intracellular concentration of either polyamine measured. We conclude that phosphatidylcholine-induced immunosuppression is not caused by methionine and polyamines are not involved in SAM-induced inhibition of TNF-alpha production. The limitation of TNF-alpha release by spermidine is specific and is not due to its conversion into spermine. PMID- 17132110 TI - Topotecan and methotrexate alter expression of the apoptosis-related genes BCL2, FAS and BCL2L12 in leukemic HL-60 cells. AB - The BCL2 family of genes (B-cell CLL/lymphoma 2; Bcl-2) plays a pivotal role in the highly regulated process of apoptosis. We have recently cloned a newly identified member of this family, BCL2L12, which was found to be differentially expressed in many tumors. It is known that topotecan and methotrexate act through induction of apoptosis in cancer cells. In the present study we investigated the expression profile of the novel apoptotic gene BCL2L12 in relation to other apoptotic genes in the human leukemic cell line HL-60, after treatment with topotecan or methotrexate. The kinetics of apoptosis induction and cell toxicity were investigated by DNA laddering and the MTT method, respectively. Gene expression levels were analyzed by RT-PCR using gene-specific primers. Downregulation of BCL2L12, BCL2 and FAS was observed after treatment of HL-60 cells with topotecan, while treatment with methotrexate led to downregulation of BCL2 and FAS, with no change in BCL2L12 expression. Our results support the significance of mRNA modulations in the expression of apoptosis-related genes during treatment of human leukemic cells with anticancer drugs. PMID- 17132111 TI - Two secreted cystatins of the soft tick Ornithodoros moubata: differential expression pattern and inhibitory specificity. AB - Two genes coding for cysteine peptidase inhibitors of the cystatin family (Om cystatin 1 and 2) were isolated from a gut-specific cDNA library of the soft tick Ornithodoros moubata. Both cystatins were clearly down-regulated after a blood meal. Om-cystatin 1 is mainly expressed in the tick gut, while Om-cystatin 2 mRNA was also found in other tick tissues. Authentic Om-cystatin 2 was significantly more abundant than Om-cystatin 1 in the gut contents of fasting ticks and was associated with hemosome-derived residual bodies accumulated in the gut lumen. Om cystatin 2 was also expressed by type 2 secretory cells in the salivary glands of unfed ticks. The inhibitory specificity of recombinant Om-cystatins 1 and 2 was tested with mammalian cysteine peptidases, as well as endogenous cysteine peptidases present in the tick gut. Both cystatins efficiently inhibited papain like peptidases, including cathepsin B and H, but differed significantly in their affinity towards cathepsin C and failed to block asparaginyl endopeptidase. Our results suggest that the secreted cystatin isoinhibitors are involved in the regulation of multiple proteolytic targets in the tick digestive system and tick host interaction. PMID- 17132116 TI - Looking back, looking forward: achievements and future directions of physiotherapy in Australia. PMID- 17132117 TI - Prospective registration of clinical trials. PMID- 17132118 TI - Strengthening interventions increase strength and improve activity after stroke: a systematic review. AB - QUESTION: Is strength training after stroke effective (ie, does it increase strength), is it harmful (ie, does it increase spasticity), and is it worthwhile (ie, does it improve activity)? DESIGN: Systematic review with meta-analysis of randomised trials. PARTICIPANTS: Stroke participants were categorised as (i) acute, very weak, (ii) acute, weak, (iii) chronic, very weak, or (iv) chronic, weak. INTERVENTION: Strengthening interventions were defined as interventions that involved attempts at repetitive, effortful muscle contractions and included biofeedback, electrical stimulation, muscle re-education, progressive resistance exercise, and mental practice. OUTCOME MEASURES: Strength was measured as continuous measures of force or torque or ordinal measures such as manual muscle tests. Spasticity was measured using the modified Ashworth Scale, a custom made scale, or the Pendulum Test. Activity was measured directly, eg, 10-m Walk Test, or the Box and Block Test, or with scales that measured dependence such as the Barthel Index. RESULTS: 21 trials were identified and 15 had data that could be included in a meta-analysis. Effect sizes were calculated as standardised mean differences since various muscles were studied and different outcome measures were used. Across all stroke participants, strengthening interventions had a small positive effect on both strength (SMD 0.33, 95% CI 0.13 to 0.54) and activity (SMD 0.32, 95% CI 0.11 to 0.53). There was very little effect on spasticity (SMD -0.13, 95% CI -0.75 to 0.50). CONCLUSION: Strengthening interventions increase strength, improve activity, and do not increase spasticity. These findings suggest that strengthening programs should be part of rehabilitation after stroke. PMID- 17132119 TI - Twelve weeks of nightly stretch does not reduce thumb web-space contractures in people with a neurological condition: a randomised controlled trial. AB - QUESTION: What is the effectiveness of 12 weeks of nightly stretch in reducing thumb web-space contracture in people with neurological conditions? DESIGN: Assessor-blinded, randomised controlled trial. PARTICIPANTS: Forty-four (one dropout)community-dwelling patients with a neurological condition (14 stroke, 7 traumatic brain injury, 23 spinal cord injury) who had uni or bilateral thumb web space contractures (60 thumbs). INTERVENTION: The experimental thumbs were splinted into a stretched,abducted position each night for 12 weeks. The control thumbs were not splinted. OUTCOME MEASURES: Thumb web-space was measured as the carpometacarpal angle during the application of a 0.9 Nm abduction torque before and after intervention. RESULTS: The mean increase in thumb web-space after 12 weeks was 1 deg (95% CI, -1 to 2). CONCLUSION: Intensive stretch administered regularly over three months does not reduce thumb web-space contractures in neurological conditions. PMID- 17132120 TI - Six sessions of manual therapy increase knee flexion and improve activity in people with anterior knee pain: a randomised controlled trial. AB - QUESTION: What are the effects of manual therapy on pain, range of motion, and activity in patients with anterior knee pain? DESIGN: Randomised controlled trial. PARTICIPANTS: Thirty-eight ambulatory care patients (one dropout) with anterior knee pain. INTERVENTION: The experimental intervention consisted of six sessions of manual therapy, while the control intervention was to remain on the waiting list for two weeks. OUTCOME MEASURES: Pain was measured using the Patellofemoral Pain Severity Questionnaire. Active knee flexion and extension was measured from photographs. Activity was measured by having the participants step up and down a 15 cm step, leading with the painful leg as many times as they could in a 60 second period. Measurements were taken before and after intervention by a blinded assessor. RESULTS: The experimental group decreased their pain by -8 mm (95% CI to 1 p =0.08) and pain on stairs by-10, (95% CI -22 to 2 p = 0.10) compared with the control group. They increased their active knee flexion by 10 deg (95% CI TO 16, p = 0.004) and and the number of steps in 60 seconds by 5 (95% CI 2 TO 8, p = 0.001) compared with the control group. CONCLUSION: Manual therapy is effective improving knee flexion and stair climbing i patients with anterior knee pain. There is a trend towards a small improvement in pain. PMID- 17132121 TI - Manual vibration increases expiratory flow rate via increased intrapleural pressure in healthy adults: an experimental study. AB - QUESTION: What is the relationship between vibration of the chest wall and the resulting chest wall force, chest wall circumference,intrapleural pressure, and expiratory flow rate? Is the change in intrapleural pressure during vibration the sum of the intrapleural pressure due to recoil of the lung, chest wall compression, and chest wall oscillation? DESIGN: Randomised, within subject,experimental study. PARTICIPANTS: Seven experienced cardiopulmonary physiotherapists and three healthy adults. INTERVENTION: Vibration (compression + oscillation), compression alone, and oscillation alone were applied manually to the chest walls of healthy participants during passive exertion and compared with passive expiration alone. OUTCOME MEASURES: Chest wall force, chest wall circumference, intrapleural pressure, and expiratory flow rate. RESULTS: During vibration, coherence was high(r2 > 0.97) between external chest wall force, chest wall circumference, intrapleural pressure, and expiratory flow. The mean change in intrapleural pressure during vibration was 9.55 cmH2O (SD 1.66), during chest compression alone was 8.06 cmH2O(SD 1.65), during oscillation alone was 7.93 cmH2O (SD 1.57), and during passive expiration alone was 6.82 cmH2O (SD 1.51). During vibration, compression contributed 13% of the change in intrapleural pressure, oscillation contributed 12%, and lung recoil contributed the remaining 75%. CONCLUSIONS: During vibration the chest behaves as a highly linear system. Changes in intrapleural pressure occurring during vibration appear to be the sum of changes in pressure due to lung recoil and the compressive and oscillatory components of the technique, which suggests that all three components are required to optimise expiratory flow. PMID- 17132122 TI - Active involvement and long-term goals influence long-term adherence to behavioural graded activity in patients with osteoarthritis: a qualitative study. AB - QUESTION: Why do some patients who have received a behavioural graded activity program successfully integrate the activities into their daily lives and others do not? DESIGN: Qualitative study. PARTICIPANTS: 12 patients were selected according to the model of deliberate sampling for heterogeneity, based on their success with the intervention as assessed on the Patient Global Assessment. INTERVENTION: Behavioural graded activity. OUTCOME MEASURES: Data from 12 interviews were coded and analysed using the methods developed in grounded theory. The interviews covered three main themes: aspects related to the content of behavioural graded activity, aspects related to experience with the physiotherapist, and aspects related to characteristics of the participant. RESULTS: Interview responses suggest that two factors influence long-term adherence to exercise and activity.First, initial long-term goals rather than short-term goals seem to relate to greater adherence to performing activities in the long term. Second, active involvement by participants in the intervention process seems to relate to greater adherence to performing activities in the long term. CONCLUSION: Although involvement of patients in the intervention process is already part of behavioural graded activity, it would be beneficial to emphasise the importance of active involvement by patients right from the start of the intervention. Furthermore, to increase the success of behavioural graded activity, physiotherapists should gain a clear understanding of the patient's initial motives in undergoing intervention. PMID- 17132123 TI - Change in impairments in the first two treatments predicts outcome in impairments, but not in activity limitations, in subacute neck pain: an observational study. AB - QUESTION: Does change in impairments within and between the first two manual therapy treatments predict change in activity limitations by the end of treatment in patients with subacute neck symptoms? DESIGN: Longitudinal, observational study. PARTICIPANTS: 29 people with neck pain for more than two weeks who subsequently received >or= three treatments. OUTCOME MEASURES: Impairments measured were active neck ROM in six directions (total ROM), most limited direction of ROM (limited ROM), pain intensity, and pain location. Activity limitations were measured using the Neck Disability Index and the Patient Specific Functional Scale. Patients' perceptions of change were measured using the Global Perceived Effect Scale. Impairments and patients' perceptions were measured before and after the first two treatments and before the final treatment whereas activity limitations were measured only before the first and last treatments. RESULTS: All measures improved by the end of treatment.Between treatment change in limited ROM predicted change in limited ROM (rs2 = 0.53 and 0.57) and total ROM (rs2 = 0.26) by the end of treatment. Within- and between treatment change in pain location predicted change in pain location (rs2 = 0.24, 0.27,0.28, and 0.57) by the end of treatment. No significant relationships were found between change in any impairments in the first two treatments and change in activity limitations by the end of treatment. CONCLUSIONS: Change in impairments predicts change in the same impairment by the end of treatment, but not in other impairments or activity limitations. It is recommended that there assessments used to guide and refine treatment be individualised and related to the specific goals for that patient. PMID- 17132124 TI - The prevalence of thumb problems in Australian physiotherapists is high: an observational study. AB - QUESTION: What is the lifetime and current prevalence of thumb problems in Australian physiotherapists and what are the factors associated with thumb problems? DESIGN: Survey of a random cross-section of physiotherapists. PARTICIPANTS: 1562(approximately 10% of the total) registered Australian physiotherapists. OUTCOME MEASURES: General questions covered demographic information, area of practice, hours worked per week, and years worked as a physiotherapist. Specific questions about thumb problems covered thumb affected, symptoms, onset of symptoms, treatment sought, relevance of work-related factors, and joint hypermobility. RESULTS: 1102 (71%) questionnaires were returned and 961 (68%) completed. The lifetime prevalence of thumb problems was 65% and the current prevalence was 41%. Factors that were significantly associated with thumb problems included: working in orthopaedic outpatients (OR 3.2, 95% CI 1.8 to 5.8); using manual therapy (OR 2.3 to 3.4, 95% CI 1.7 to 5.1), trigger point therapy (OR 2.3, 95% CI 1.7 to 3.0) and massage (OR 2.1, 95% CI 1.6 to 2.8); having thumb joint hypermobility (OR 2.2 to 2.6, 95% CI 1.4 to 4.5); or an inability to stabilise the joints of the thumb whilst performing physiotherapy techniques (OR 4.2, 95% CI 2.9 to 5.9). Of those respondents who reported thumb problems, 19% had changed their area of practice and 4% had left the profession as a result of their thumb problems. CONCLUSION: The prevalence of thumb problems in Australian physiotherapists appears to be high and can be of sufficient severity to impact on careers. PMID- 17132125 TI - Compliance with the guidelines for acute ankle sprain for physiotherapists is moderate in the Netherlands: an observational study. AB - QUESTION: What is the compliance with guidelines for acute ankle sprain for physiotherapists? DESIGN: Survey of random sample of physiotherapists. PARTICIPANTS: 400 physiotherapists working in extramural health care in the Netherlands. OUTCOME MEASURES: Questions covered attitude towards guidelines in general, familiarity with the guidelines for acute ankle sprain,compliance with the guidelines, advantages and disadvantages of the guidelines, and factors relating to compliance with the guidelines. RESULTS: The majority of the physiotherapists were familiar with the content of the guidelines to some degree and 66%applied it to more than half of their patients with acute ankle sprain. The recommendations to determine both the prognosis and the necessity of treatment by using the function score were the least followed. Some physiotherapists thought the function score was not completely clear, which may have been a barrier for implementation. Factors relating positively to compliance were a positive attitude towards guidelines in general, and having colleagues who implemented the guidelines for acute ankle sprain. CONCLUSION: Although compliance with the guidelines for acute ankle sprain was fair/moderate, compliance may be enhanced by improving clarity of the function score, including it in the short version and improving the attitude of physiotherapists towards guidelines in general. PMID- 17132126 TI - Weight training does not promote lymphoedema in breast cancer survivors. PMID- 17132127 TI - The effect of 300 mW, 830 nm laser on chronic neck pain. PMID- 17132128 TI - The presence of four simple history features can diagnose migraine accurately. PMID- 17132129 TI - In addition to usual care, pelvic floor exercises commenced preoperatively reduce incontinence after prostatectomy. PMID- 17132130 TI - Spinal manipulation and exercise was better than ultrasound and exercise for patients with chronic low back pain. PMID- 17132131 TI - Pelvic floor muscle strength testing. PMID- 17132132 TI - Clinical diagnostic tests for the sacroiliac joint: motion and palpation tests. PMID- 17132137 TI - (Peri)vascular production and action of pro-inflammatory cytokines in brain pathology. AB - In response to tissue injury or infection, the peripheral tissue macrophage induces an inflammatory response through the release of IL-1beta (interleukin 1beta) and TNFalpha (tumour necrosis factor alpha). These cytokines stimulate macrophages and endothelial cells to express chemokines and adhesion molecules that attract leucocytes into the peripheral site of injury or infection. The aims of the present review are to (i) discuss the relevance of brain (peri)vascular cells and compartments to bacterial meningitis, HIV-1-associated dementia, multiple sclerosis, ischaemic and traumatic brain injury, and Alzheimer's disease, and (ii) to provide an overview of the production and action of pro inflammatory cytokines by (peri)vascular cells in these pathologies of the CNS (central nervous system). The brain (peri)vascular compartments are highly relevant to pathologies affecting the CNS, as infections are almost exclusively blood-borne. Insults disrupt blood and energy flow to neurons, and active brain to-blood transport mechanisms, which are the bottleneck in the clearance of unwanted molecules from the brain. Perivascular macrophages are the most reactive cell type and produce IL-1beta and TNFalpha after infection or injury to the CNS. The main cellular target for IL-1beta and TNFalpha produced in the brain (peri)vascular compartment is the endothelium, where these cytokines induce the expression of adhesion molecules and promote leucocyte infiltration. Whether this and other effects of IL-1 and TNF in the brain (peri)vascular compartments are detrimental or beneficial in neuropathology remains to be shown and requires a clear understanding of the role of these cytokines in both damaging and repair processes in the CNS. PMID- 17132138 TI - Life and death decisions of the pancreatic beta-cell: the role of fatty acids. AB - Both stimulatory and detrimental effects of NEFAs (non-esterified fatty acids) on pancreatic beta-cells have been recognized. Acute exposure of the pancreatic beta cell to high glucose concentrations and/or saturated NEFAs results in a substantial increase in insulin release, whereas chronic exposure results in desensitization and suppression of secretion, followed by induction of apoptosis. Some unsaturated NEFAs also promote insulin release acutely, but they are less toxic to beta-cells during chronic exposure and can even exert positive protective effects. Therefore changes in the levels of NEFAs are likely to be important for the regulation of beta-cell function and viability under physiological conditions. In addition, the switching between endogenous fatty acid synthesis or oxidation in the beta-cell, together with alterations in neutral lipid accumulation, may have critical implications for beta-cell function and integrity. Long-chain acyl-CoA (formed from either endogenously synthesized or exogenous fatty acids) controls several aspects of beta-cell function, including activation of specific isoenzymes of PKC (protein kinase C), modulation of ion channels, protein acylation, ceramide formation and/or NO-mediated apoptosis, and transcription factor activity. In this review, we describe the effects of exogenous and endogenous fatty acids on beta-cell metabolism and gene and protein expression, and have explored the outcomes with respect to insulin secretion and beta-cell integrity. PMID- 17132139 TI - Endoplasmic reticulum chaperones inhibit the production of amyloid-beta peptides. AB - Abeta (amyloid-beta peptides) generated by proteolysis of APP (beta-amyloid precursor protein), play an important role in the pathogenesis of AD (Alzheimer's disease). ER (endoplasmic reticulum) chaperones, such as GRP78 (glucose-regulated protein 78), make a major contribution to protein quality control in the ER. In the present study, we examined the effect of overexpression of various ER chaperones on the production of Abeta in cultured cells, which produce a mutant type of APP (APPsw). Overexpression of GRP78 or inhibition of its basal expression, decreased and increased respectively the level of Abeta40 and Abeta42 in conditioned medium. Co-expression of GRP78's co-chaperones ERdj3 or ERdj4 stimulated this inhibitory effect of GRP78. In the case of the other ER chaperones, overexpression of some (150 kDa oxygen-regulated protein and calnexin) but not others (GRP94 and calreticulin) suppressed the production of Abeta. These results indicate that certain ER chaperones are effective suppressors of Abeta production and that non-toxic inducers of ER chaperones may be therapeutically beneficial for AD treatment. GRP78 was co-immunoprecipitated with APP and overexpression of GRP78 inhibited the maturation of APP, suggesting that GRP78 binds directly to APP and inhibits its maturation, resulting in suppression of the proteolysis of APP. On the other hand, overproduction of APPsw or addition of synthetic Abeta42 caused up-regulation of the mRNA of various ER chaperones in cells. Furthermore, in the cortex and hippocampus of transgenic mice expressing APPsw, the mRNA of some ER chaperones was up-regulated in comparison with wild-type mice. We consider that this up-regulation is a cellular protective response against Abeta. PMID- 17132140 TI - Antimicrobial peptides and proteins, exercise and innate mucosal immunity. AB - This review examines the question of whether exercise can be used as an experimental model to further our understanding of innate antimicrobial peptides and proteins (AMPs) and their role in susceptibility to infection at mucosal surfaces. There is strong evidence to suggest that AMPs, in combination with cellular and physical factors, play an important role in preventing infection. Although AMPs act directly on microorganisms, there is increasing recognition that they also exert their protective effect via immunomodulatory mechanisms, especially in noninflammatory conditions. Further studies that manipulate physiologically relevant concentrations of AMPs are required to shed light on the role they play in reducing susceptibility to infection. Evidence shows that in various form prolonged and/or exhaustive exercise is a potent modulator of the immune system, which can either sharpen or blunt the immune response to pathogens. The intensity and duration of exercise can be readily controlled in experimental settings to manipulate the degree of physical stress. This would allow for an investigation into a potential dose-response effect between exercise and AMPs. In addition, the use of controlled exercise could provide an experimental model by which to examine whether changes in the concentration of AMPs alters susceptibility to illness. PMID- 17132141 TI - Detection of four lymphotropic herpesviruses in Hungarian patients with multiple myeloma and lymphoma. AB - It has been suggested that human herpesvirus 8 (HHV-8), also known as KSHV (Kaposi's sarcoma-associated human herpesvirus), might possess a promoting effect in the development and progression of monoclonal gammopathies. In this study, the presence of Epstein-Barr virus (EBV), human cytomegalovirus (CMV), human herpesvirus 6 (HHV-6) and human herpesvirus 8 (HHV-8) were tested in patients with multiple myeloma (MM) using both serologic and nucleic acid amplification techniques. The transient reactivation or continuous presence of EBV, CMV, HHV-6 and HHV-8 could be detected in, respectively, 36, eight, 13 and 29 of 69 MM patients; nine, one, four and six of 16 monoclonal gammopathy of unknown significance patients; and seven, four, zero and five of 10 Waldenstrom's macroglobulinemia patients. The total number of MM patients was 95. HHV-8 PCR positivity was significantly more frequent in the MM group than in the control group of patients with non-Hodgkin's lymphoma (NHL). However, serologic testing did not reveal significant differences between the two patient groups. The number of MM patients with concomitant herpesvirus infections as detected by PCR was as follows: 15 double, seven triple and two quadruple virus nucleic acid positive. In 13/95 MM patients, the simultaneous presence of acute EBV infection and HHV-8 PCR-positivity was detected compared with none of the control group (P=0.009). These results indicate that in addition to HHV-8, the transitional reactivation of EBV may also play a role in the pathogenesis of MM. PMID- 17132142 TI - Metabolic flux analysis of a glycerol-overproducing Saccharomyces cerevisiae strain based on GC-MS, LC-MS and NMR-derived C-labelling data. AB - This study focuses on unravelling the carbon and redox metabolism of a previously developed glycerol-overproducing Saccharomyces cerevisiae strain with deletions in the structural genes encoding triosephosphate isomerase (TPI1), the external mitochondrial NADH dehydrogenases (NDE1 and NDE2) and the respiratory chain linked glycerol-3-phosphate dehydrogenase (GUT2). Two methods were used for analysis of metabolic fluxes: metabolite balancing and (13)C-labelling-based metabolic flux analysis. The isotopic enrichment of intracellular primary metabolites was measured both directly (liquid chromatography-MS) and indirectly through proteinogenic amino acids (nuclear magnetic resonance and gas chromatography-MS). Because flux sensitivity around several important metabolic nodes proved to be dependent on the applied technique, the combination of the three (13)C quantification techniques generated the most accurate overall flux pattern. When combined, the measured conversion rates and (13)C-labelling data provided evidence that a combination of assimilatory metabolism and pentose phosphate pathway activity diverted some of the carbon away from glycerol formation. Metabolite balancing indicated that this results in excess cytosolic NADH, suggesting the presence of a cytosolic NADH sink in addition to those that were deleted. The exchange flux of four-carbon dicarboxylic acids across the mitochondrial membrane, as measured by the (13)C-labelling data, supports a possible role of a malate/aspartate or malate/oxaloacetate redox shuttle in the transfer of these redox equivalents from the cytosol to the mitochondrial matrix. PMID- 17132143 TI - Early transcriptional response of wine yeast after rehydration: osmotic shock and metabolic activation. AB - The inoculation of active dry wine yeast (ADWY) is one of the most common practices in winemaking. We have used DNA microarray technology to examine the genetic expression patterns of a commercial ADWY strain after rehydration. After rehydration of ADWY for 30 min, a further hour in water after rehydration did not lead to any relevant changes in global gene expression. Expression changes in rehydrated cells upon incubation in a sorbitol solution at the same osmotic pressure as in complete must were rather limited, whereas the presence of fermentable carbon sources or the complete medium (synthetic must) produced very similar transcriptional responses. The main responses were the activation of some genes of the fermentation pathway and of the nonoxidative branch of the pentose phosphate pathway, and the induction of a huge cluster of genes related to ribosomal biogenesis and protein synthesis. The presence of cycloheximide in fermentable medium produced a similar but stronger transcriptional response. Whereas the viabilities of rehydrated cells incubated for 1 h in these different media were similar, yeast vitality, which represents the fermentative capacity of the yeast, showed a positive correlation with the availability of a fermentable carbon source. PMID- 17132144 TI - Targeting of TMV movement protein to plasmodesmata requires the actin/ER network: evidence from FRAP. AB - Fluorescence recovery after photobleaching (FRAP) was used to study the mechanism by which fluorescent-protein-tagged movement protein (MP) of tobacco mosaic virus (TMV) is targeted to plasmodesmata (PD). The data show that fluorescence recovery in PD at the leading edge of an infection requires elements of the cortical actin/endoplasmic reticulum (ER) network and can occur in the absence of an intact microtubule (MT) cytoskeleton. Inhibitors of the actin cytoskeleton (latrunculin and cytochalasin) significantly inhibited MP targeting, while MT inhibitors (colchicine and oryzalin) did not. Application of sodium azide to infected cells implicated an active component of MP transfer to PD. Treatment of cells with Brefeldin A (BFA) at a concentration that caused reabsorption of the Golgi bodies into the ER (precluding secretion of viral MP) had no effect on MP targeting, while disruption of the cortical ER with higher concentrations of BFA caused significant inhibition. Our results support a model of TMV MP function in which targeting of MP to PD during infection is mediated by the actin/ER network. PMID- 17132145 TI - Nuclear export of influenza A virus mRNAs requires ongoing RNA polymerase II activity. AB - Influenza A virus transcribes its segmented negative sense RNA genome in the nuclei of infected cells in a process long known to require host RNA polymerase II (RNAP-II). RNA polymerase II synthesizes pre-mRNAs whose 5'-cap structures are scavenged by the viral RNA-dependent RNA polymerase during synthesis of viral mRNAs. Drugs that inhibit RNAP-II therefore block viral replication, but not necessarily solely by denying the viral polymerase a source of cap-donor molecules. We show here that 5,6-dichloro-1-beta-D-ribofuranosyl-benzimidazole (DRB), a compound that prevents processive transcription by RNAP-II, inhibits expression of the viral HA, M1 and NS1 genes at the post-transcriptional level. Abundant quantities of functionally and structurally intact viral mRNAs are made in the presence of DRB but with the exception of NP and NS2 mRNAs, are not efficiently translated. Taking M1 and NP mRNAs as representatives of DRB sensitive and insensitive mRNAs, respectively, we found that the block to translation operates at the level of nuclear export. Furthermore, removal of DRB reversed this block unless a variety of chemically and mechanistically distinct RNAP-II inhibitors were added instead. We conclude that influenza A virus replication requires RNAP-II activity not just to provide capped mRNA substrates but also to facilitate nuclear export of selected viral mRNAs. PMID- 17132146 TI - Rab10 is involved in basolateral transport in polarized Madin-Darby canine kidney cells. AB - The sorting of newly synthesized membrane proteins to the cell surface is an important mechanism of cell polarity. To identify more of the molecular machinery involved, we investigated the function of the small GTPase Rab10 in polarized epithelial Madin-Darby canine kidney cells. We find that GFP-tagged Rab10 localizes primarily to the Golgi during early cell polarization. Expression of an activated Rab10 mutant inhibits biosynthetic transport from the Golgi and missorts basolateral cargo to the apical membrane. Depletion of Rab10 by RNA interference has only mild effects on biosynthetic transport and epithelial polarization, but simultaneous inhibition of Rab10 and Rab8a more strongly impairs basolateral sorting. These results indicate that Rab10 functions in trafficking from the Golgi at early stages of epithelial polarization, is involved in biosynthetic transport to the basolateral membrane and may co-operate with Rab8. PMID- 17132147 TI - Patients with a non-dysferlin Miyoshi myopathy have a novel membrane repair defect. AB - Two autosomal recessive muscle diseases, limb girdle muscular dystrophy type 2B (LGMD2B) and Miyoshi myopathy (MM), are caused by mutations in the dysferlin gene. These mutations result in poor ability to repair cell membrane damage, which is suggested to be the cause for this disease. However, many patients who share clinical features with MM-type muscular dystrophy do not carry mutations in dysferlin gene. To understand the basis of MM that is not due to mutations in dysferlin gene, we analyzed cells from patients in one such family. In these patients, we found no defects in several potential candidates - annexin A2, caveolin-3, myoferlin and the MMD2 locus on chromosome 10p. Similar to dysferlinopathy, these cells also exhibit membrane repair defects and the severity of the defect correlated with severity of their disease. However, unlike dysferlinopathy, none of the conventional membrane repair pathways are defective in these patient cells. These results add to the existing evidence that cell membrane repair defect may be responsible for MM-type muscular dystrophy and indicate that a previously unsuspected genetic lesion that affects cell membrane repair pathway is responsible for the disease in the non-dysferlin MM patients. PMID- 17132148 TI - A CDPK isoform participates in the regulation of nodule number in Medicago truncatula. AB - Medicago spp. are able to develop root nodules via symbiotic interaction with Sinorhizobium meliloti. Calcium-dependent protein kinases (CDPKs) are involved in various signalling pathways in plants, and we found that expression of MtCPK3, a CDPK isoform present in roots of the model legume Medicago truncatula, is regulated during the nodulation process. Early inductions were detected 15 min and 3-4 days post-inoculation (dpi). The very early induction of CPK3 messengers was also present in inoculated M. truncatula dmi mutants and in wild-type roots subjected to salt stress, indicating that this rapid response is probably stress related. In contrast, the later response was concomitant with cortical cell division and the formation of nodule primordia, and was not observed in wild-type roots inoculated with nod (-) strains. This late induction correlated with a change in the subcellular distribution of CDPK activities. Accordingly, an anti MtCPK3 antibody detected two bands in soluble root extracts and one in the particulate fraction. CPK3::GFP fusions are targeted to the plasma membrane in epidermal onion cells, a localization that depends on myristoylation and palmitoylation sites of the protein, suggesting a dual subcellular localization. MtCPK3 mRNA and protein were also up-regulated by cytokinin treatment, a hormone linked to the regulation of cortical cell division and other nodulation-related responses. An RNAi-CDPK construction was used to silence CPK3 in Agrobacterium rhizogenes-transformed roots. Although no major phenotype was detected in these roots, when infected with rhizobia, the total number of nodules was, on average, twofold higher than in controls. This correlates with the lack of MtCPK3 induction in the inoculated super-nodulator sunn mutant. Our results suggest that CPK3 participates in the regulation of the symbiotic interaction. PMID- 17132149 TI - Impact of conjugal transfer on the stability of IncP-1 plasmid pKJK5 in bacterial populations. AB - The intrinsic stability of IncP-1 plasmid pKJK5 was assessed in both an Escherichia coli and a Kluyvera sp. population maintained in bacterial mats and in liquid nutrient broth without selective pressure. A fluorescence tagging/flow cytometry approach was used to detect and quantify plasmid loss from populations harboring either conjugation-proficient or -deficient pKJK5 derivatives. The results show that the plasmid's ability to conjugate plays an important role in its stable maintenance in populations of both species. This effect was most pronounced in dense bacterial populations and to a far lesser extent during growth in liquid broth. Furthermore, conjugation-proficient plasmids were able to spread infectiously in the bacterial mats initiated with various ratios of plasmid-harboring cells, resulting in a nearly exclusively plasmid-harboring population. PMID- 17132150 TI - Binding study of AfsK, a Ser/Thr kinase from Streptomyces coelicolor A3(2) and S adenosyl-L-methionine. AB - Streptomyces coelicolor A3(2) produces an antibiotic, actinorhodin, which belongs to the aromatic polyketides and which can function as an acid/base indicator. Its production results in the death of microorganisms in the vicinity of S. coelicolor A3(2), and this phenomenon can be used in concert with biopesticides. The exogenous addition of S-adenosyl-L-methionine (SAM) to S. coelicolor A3(2) enhances its actinorhodin production and may initiate actinorhodin biosynthesis, with at least four genes being involved. Of these (because afsK initiates the others), AfsK, the protein expressed from afsK, may be interacting with SAM. Although the three-dimensional structure of AfsK has not been determined, the differences between nuclear magnetic resonance (NMR) signals obtained from the free form of SAM and those from a SAM-protein complex can help us to determine whether SAM binds to the C-terminal of AfsK or not. In the present study, NMR data analysis strongly supported the idea that SAM binds to AfsK. PMID- 17132151 TI - Interaction of Bacillus thuringiensis svar. israelensis Cry toxins with binding sites from Aedes aegypti (Diptera: Culicidae) larvae midgut. AB - This work shows in vitro processing of Bacillus thuringiensis svar. isralensis Cry toxins and the capacity of the active fragments to bind the midgut microvilli of Aedes aegypti larvae. Processing of Cry11Aa, Cry4Aa and Cry4Ba yielded double fragments of 38-30, 45-20 and 45-18 kDa, respectively. Competition assays showed that all active (125)I-Cry toxins are able to specifically bind to brush border membrane fractions and they might share a common class of binding sites. The values of IC(50) suggested that toxins do not display high affinity for the receptors from brush border membrane fractions, while dissociation assays showed that binding was irreversible, indicating the insertion of toxins in the cell membrane. PMID- 17132152 TI - Selective impairment of noradrenergic transmission in the bisected rat vas deferens following photochemically-induced cerebral ischaemia. AB - BACKGROUND: Cerebrovascular disease may impair the autonomic control of peripheral organs including the male urogenital tract. This study investigates the effect of cortico-parietal focal ischaemia on the adrenergic and purinergic transmission in isolated epididymal and prostatic portion of rat vas deferens. METHODS: Focal brain ischaemia was induced in male rats by photochemical activation following rose bengal intravenous injection. Twenty-four hours following brain ischaemia, cumulative and non-cumulative concentration-response curves were obtained for noradrenaline and alpha,beta-methylene ATP in the right and left epididymal and prostatic portions of the vas deferens. Both portions were also stimulated by single-pulse or pulse trains at 2-30 Hz to produce isometric contractions. RESULTS: In both portions from ischaemic rats the response to exogenous noradrenaline was markedly depressed compared with controls. Acute cortico-parietal ischaemia almost completely abolished the adrenergic phase of the response to single-pulse stimulation in the epididymal portion of the vas deferens. In addition, brain ischaemia deeply depressed phasic and tonic contractions of the frequency-response curve in both portions of bisected vas deferens. CONCLUSIONS: Cortico-parietal ischaemia produces a selective noradrenergic impairment at the level of male sexual secondary organs that may contribute to sexual dysfunction after stroke. PMID- 17132153 TI - Male serum Chlamydia trachomatis IgA and IgG, but not heat shock protein 60 IgG, correlates with negatively affected semen characteristics and lower pregnancy rates in the infertile couple. AB - BACKGROUND: The objective of this study was to evaluate whether serum Chlamydia trachomatis immunoglobulin-A (IgA), IgM and C. trachomatis heat shock protein 60 (CHSP60) IgG are of additional value to C. trachomatis IgG regarding the impact on fecundity in infertile couples, and to relate C. trachomatis serum antibodies to semen characteristics, diagnoses and pregnancy outcome. METHODS: A total of 226 infertile couples, previously tested for C. trachomatis IgG, were tested for C. trachomatis IgA, IgM and CHSP60 IgG, and semen samples from all men were analysed. RESULTS: Chlamydia trachomatis serum IgA in men (but not in women) correlated with reduced chances of achieving pregnancy [p = 0.021, relative risk (RR) =0.65, 95% confidence interval (CI) 0.42-1.005] and in combination with C. trachomatis IgG the chance was further reduced (p =0.001, RR = 0.35, 95% CI 0.15 0.84). Chlamydia trachomatis serum IgA was also significantly correlated with reduced motility of the spermatozoa (-8.7%, p = 0.023), increased number of dead spermatozoa (+10.5%, p = 0.014) and higher prevalence of leucocytes in semen (+122%, p = 0.005), and in combination with C. trachomatis IgG positivity, there was also a decrease in sperm concentration (-35%, p = 0.033), the number of progressive spermatozoa (-14.8%, p = 0.029) and a rise in the teratozoospermia index (+4.4%, p = 0.010). CHSP60 IgG correlated with reduced motility (-5.6%, p = 0.033), and in the women to tubal factor infertility (p = 0.033), but no correlations of C. trachomatis serum IgM or CHSP60 IgG with pregnancy rates were found. CONCLUSIONS: Chlamydia trachomatis serum IgA in the male partner of the infertile couple has an additive value to IgG in predicting pregnancy chances, and serum IgA and IgG are associated with subtle negative changes in semen characteristics. PMID- 17132154 TI - Modifications of surface carbohydrates on bovine spermatozoa mediated by oviductal fluid: a flow cytometric study using lectins. AB - The objective of the present study was to characterize and quantify changes in exposed saccharide residues of bovine sperm during capacitation in oviductal fluid (ODF) using flow cytometry (FC). Bovine sperm were incubated with 0% or 50% non-luteal ODF for 30 min or 3.5 h. After incubation, sperm were labelled with 11 fluorescein isothiocyanate-labelled lectins and evaluated for lectin binding with FC. Furthermore, inhibiting sugars were used to determine specificity of lectin binding to oligosaccharides on the sperm surface. After 30 min incubation, there was a 91% decrease in fluorescence intensity of labelled sperm incubated in WGA, a 76% decline for Con A, 75% decline for BS-I and a 36% decline for DBA. These differences remained approximately the same over the 3.5-h incubation. Interestingly, although there was no reduction in UEA-I binding at 30 min, a significant reduction (23%) was observed at 3.5 h. Con A fluorescence was mostly inhibited with either alpha-d-glucose or alpha-d-mannose (86% and 90% respectively). BS-I fluorescence was reduced after prior incubation of the control samples with N-acetyl-galactosamine and galactose by 74% and 80% respectively. After prior incubation with N-acetyl-galactosamine DBA fluorescence reduced by 18% in the control samples. With UEA-I no fluorescence reduction was observed after prior incubation with l-fucose. We have demonstrated that capacitation of bovine sperm in ODF is accompanied by a quantitative reduction in individual lectin binding sites. These modifications may be crucial to the subsequent signalling events involved with sperm-zona binding, zona penetration or interaction with the oolema. PMID- 17132155 TI - Expression of galectin-3 and its regulation in the testes. AB - Although spermatogenesis is a complex process under hormonal control, which includes mainly follicle stimulating hormone (FSH) and androgens, little is known about the intra-testicular mediators of these hormones. In the present study, galectin-3 (Gal-3) expression has been identified in human, rat and porcine testes where it is under hormonal control. Gal-3 is present in Sertoli cells and appears to be absent in human and (probably) in rat germ cells. Gal-3 expression was evidenced in the testes, in terms of both mRNA and protein (31 kDa). Gal-3 expression in cultured porcine Sertoli cells was shown to be under the positive control of FSH as well as of two cytokines epidermal growth factor (EGF) and tumour necrosis factor-alpha (TNF-alpha). Gal-3 expression in Sertoli cells is also potentially under the control of mature germ cells as an increased expression was observed in adult rat testes depleted in spermatocytes or spermatids. Although the function of testicular Gal-3 remains to be investigated, a potential role of Gal-3 in germ cell survival/regeneration is suggested based on its increased expression 1 month after a transient germ cell death process triggered by 10 days of treatment with the antiandrogen flutamide. Finally, although in the normal human testes, Gal-3 is exclusively located in the Sertoli cell cytoplasm, a nuclear localization is observed in the infertile testes. Together, the present findings have shown that (i) Gal-3 is expressed in the porcine, rat and human Sertoli cells; (ii) Gal-3 is under the positive control of FSH as well as of EGF and TNF-alpha and possibly of adult germ cells. These observations are compatible with a potential pro-survival role of Gal-3 in the testes. PMID- 17132156 TI - Evaluation of selective calculus removal by a fluorescence feedback-controlled Er:YAG laser in vitro. AB - OBJECTIVES: To evaluate the removal of subgingival calculus and dental hard tissues depending on the threshold level of a fluorescence feedback-controlled Er:YAG laser. MATERIAL AND METHODS: Twenty teeth with calculus on the root surface were treated with an Er:YAG laser. Laser settings were 140 mJ and 10 Hz. The initial fluorescence threshold level of 5 [U] was reduced at intervals of 1 [U] for every laser treatment. Areas of residual calculus (RC) were evaluated using a surface analysis software. Loss of dental hard tissues was assessed by histomorphometric analysis of undecalcified ground sections. RESULTS: Using a threshold value of 5 [U], the median amount of RC was 11% (0-78%). By lowering the threshold levels, the amount of RC decreased [level 1 [U]: 0% (0-26%)]. The laser-treated root surfaces revealed a statistically significant reduction of the cementum thickness [median: 80 microm (0-250)] compared with the non-treated opposite side [median: 90 microm (30-250)] (p<0.05). CONCLUSION: The amount of RC following laser irradiation depends on the fluorescence threshold level for a feedback-controlled Er:YAG laser. It might be suggested that this laser system may be used with a threshold level even lower than 5 [U] without removing a clinically relevant amount of root cementum. PMID- 17132157 TI - Allochthonous inputs of riverine picocyanobacteria to coastal waters in the Arctic Ocean. AB - The observed onset of climate change at high northern latitudes has highlighted the need to establish current baseline conditions in the Arctic Ocean, and has raised concern about the potential for the invasion and growth of biota that have warm temperature optima, such as cyanobacteria. In this study, we used 16S rRNA gene sequences as a molecular marker to evaluate the hypothesis that Arctic rivers provide a major inoculum of cyanobacteria into the coastal Arctic Ocean. Surface samples were collected along a transect extending from the Mackenzie River (Northwest Territories, Canada), across its estuary, to 200 km offshore at the edge of the perennial Arctic pack ice (Beaufort Sea). The highest picocyanobacteria concentrations occurred in the river, with concentrations an order of magnitude lower at offshore marine stations. The 16S rRNA gene clone libraries of five surface samples and five strains along this gradient showed that the cyanobacterial sequences were divided into eight operational taxonomic units (OTUs), six OTUs closely related to freshwater and brackish Synechococcus and two OTUs of filamentous cyanobacteria. No typically marine Synechococcus sequences and no Prochlorococcus sequences were recovered. These results are consistent with the hypothesis of an allochthonous origin of picocyanobacteria in the coastal Arctic Ocean, and imply survival but little net growth of picocyanobacteria under the present conditions in northern high-latitude seas. PMID- 17132158 TI - Diffusion tensor imaging of frontal lobe white matter tracts in schizophrenia. AB - We acquired diffusion tensor and structural MRI images on 103 patients with schizophrenia and 41 age-matched normal controls. The vector data was used to trace tracts from a region of interest in the anterior limb of the internal capsule to the prefrontal cortex. Patients with schizophrenia had tract paths that were significantly shorter in length from the center of internal capsule to prefrontal white matter. These tracts, the anterior thalamic radiations, are important in frontal-striatal-thalamic pathways. These results are consistent with findings of smaller size of the anterior limb of the internal capsule in patients with schizophrenia, diffusion tensor anisotropy decreases in frontal white matter in schizophrenia and hypothesized disruption of the frontal-striatal thalamic pathway system. PMID- 17132160 TI - Selection for early and late adult emergence alters the rate of pre-adult development in Drosophila melanogaster. AB - BACKGROUND: Circadian clocks have been implicated in the regulation of pre-adult development of fruit flies Drosophila melanogaster. It is believed that faster clocks speed up development and slower clocks slow it down. We established three sets of D. melanogaster populations (early, control and late). The early and late populations were raised by selecting for flies that emerged either in the morning or in the evening under 12:12 hr light/dark (LD) cycles. After 75 generations of selection, the time course and waveform of the adult emergence and activity rhythms of the early and the late populations diverged from each other as well as from the controls. In this paper, we report the consequence of this selection on the rate of pre-adult development. RESULTS: We assayed the pre-adult development time of the selected and control populations under 12:12 hr LD cycles and constant darkness (DD). Under LD cycles, the early populations develop faster than the controls, while the late populations develop slower than the controls. Although flies take longer to develop under DD than in LD, the relative differences between the mean development times of the selected and control populations remain unaltered in DD. In a separate experiment designed to investigate the effect of time of egg collection and experimental conditions on the duration of pre-adult stage, we assayed the development time of the selected and control populations by collecting eggs at different times of the day (morning and evening) and by assaying their pre-adult development time under constant light (LL), LD, and DD conditions. Irrespective of the time of egg collection and assay light regime, the late flies continue to develop slower than the early flies. CONCLUSION: The results of our study clearly indicate that selection on the timing of adult emergence alters the rate of pre-adult development in D. melanogaster. The timing of egg collection as well as assay light regime does not have any measurable effect on the relative differences between the developmental rates of the early and the late flies. Taken together these results appear to suggest that pleiotropic effects of clock genes mediate correlated changes in the timing of adult emergence and the rate of pre-adult development in D. melanogaster. PMID- 17132159 TI - Comprehensive analysis of the ATM, CHEK2 and ERBB2 genes in relation to breast tumour characteristics and survival: a population-based case-control and follow up study. AB - BACKGROUND: Mutations in the ataxia-telangiectasia mutated (ATM) and checkpoint kinase 2 (CHEK2) genes and amplification of the v-erb-b2 avian erythroblastic leukemia viral oncogene homolog 2 (ERBB2) gene have been suggested to have an important role in breast cancer aetiology. However, whether common variation in these genes has a role in the development of breast cancer or breast cancer survival in humans is still not clear. METHODS: We performed a comprehensive haplotype analysis of the ATM, CHEK2 and ERBB2 genes in a Swedish population based study, which included 1,579 breast cancer cases and 1,516 controls. We followed the cases for 8.5 years, on average, and retrieved information on the date and cause of death during that period from the nationwide Swedish causes of death registry. We selected seven haplotype-tagging SNPs (tagSNPs) in the ATM gene, six tagSNPs in the CHEK2 gene and seven tagSNPs in the ERBB2 gene that predicted both haplotypic and single locus variations in the respective genes with R2 values > or = 0.8. These tagSNPs were genotyped in the complete set of cases and controls. We computed expected haplotype dosages of the tagSNP haplotypes and included the dosages as explanatory variables in Cox proportional hazards or logistic regression models. RESULTS: We found no association between any genetic variation in the ATM, CHEK2 or ERBB2 genes and breast cancer survival or the risk of developing tumours with certain characteristics. CONCLUSION: Our results indicate that common variants in the ATM, CHEK2 or ERBB2 genes are not involved in modifying breast cancer survival or the risk of tumour-characteristic defined breast cancer. PMID- 17132161 TI - The relationship between parental socio-economic status and episodes of drunkenness among adolescents: findings from a cross-national survey. AB - BACKGROUND: Behavioral factors such as (excessive) alcohol consumption play a major role in the explanation of social inequalities in health. The unequal distribution of health risk behaviors among socio-economic groups has important consequences for both the current and future health status of the younger generation. However, little is known about socio-economic differences in unhealthy lifestyles during adolescence. The purpose of the present study is to investigate socio-economic differences in adolescent drinking behaviour among 11 15 year old adolescents in Europe and North America. METHODS: Data was obtained from the Health Behaviour in School-aged Children (HBSC) study 2001/02, a cross national survey conducted in collaboration with the World Health Organization. The present analysis is based on 69249 male and 73619 female students from 28 countries. The effect of parental occupation and family affluence on episodes of drunkenness was assessed using separate logistic regression models controlling for age. RESULTS: Socio-economic circumstances of the family had only a limited effect on repeated drunkenness in adolescence. For girls only in one out of 28 countries a significant association between family affluence and repeated drunkenness was observed, while boys from low and/or medium affluent families in nine countries faced a lower risk of drunkenness than boys from more affluent families. Regarding parental occupation, significant differences in episodes of drunkenness were found in nine countries for boys and in six countries for girls. Compared to family affluence, which was positively related to risk of drunkenness, a decreasing occupational status predicted an increasing risk of drunkenness. This pattern was identified within a number of countries, most noticeably for boys. CONCLUSION: Parental socio-economic status is only of limited importance for episodes of drunkenness in early adolescence, and this very limited role seems to apply for girls more than for boys and for parental occupation more than family affluence. For future studies it might be important to look at the effects of socio-economic status within the context of other peer, family and school related factors in order to assess to what extent those factors might mediate the effects of social class background. PMID- 17132162 TI - DNA unwinding assay using streptavidin-bound oligonucleotides. AB - BACKGROUND: Helicases play essential roles in many cellular processes including replication, transcription and translation. Most helicases translocate along one strand of the duplex while displacing the complementary strand (of either DNA or RNA). Thus, helicases have directionality. They move along nucleic acids in either the 3'--> 5' or 5'--> 3' direction. The directionality of helicases with low activity or of those that cannot initiate duplex unwinding from a substrate that contains only one single-stranded overhang region is difficult to determine. RESULTS: An improved assay to determine helicase directionality was developed that uses a substrate containing biotinylated oligonucleotides. As a proof of concept, it was shown that the substrates substantially improve helicase activity and directionality determination for several DNA helicases in comparison to more traditional substrates. In addition, a universal substrate that can be used to determine the directionality of both 3'--> 5' and 5'--> 3' helicases was developed. CONCLUSION: It is shown here that the use of a biotin-streptavidin complex as a helicase substrate improves helicase activity and the determination of helicase directionality. The method described is simpler that the currently available techniques. PMID- 17132163 TI - Biochemical characterisation of LigN, an NAD+-dependent DNA ligase from the halophilic euryarchaeon Haloferax volcanii that displays maximal in vitro activity at high salt concentrations. AB - BACKGROUND: DNA ligases are required for DNA strand joining in all forms of cellular life. NAD+-dependent DNA ligases are found primarily in eubacteria but also in some eukaryotic viruses, bacteriophage and archaea. Among the archaeal NAD+-dependent DNA ligases is the LigN enzyme of the halophilic euryarchaeon Haloferax volcanii, the gene for which was apparently acquired by Hfx. volcanii through lateral gene transfer (LGT) from a halophilic eubacterium. Genetic studies show that the LGT-acquired LigN enzyme shares an essential function with the native Hfx. volcanii ATP-dependent DNA ligase protein LigA. RESULTS: To characterise the enzymatic properties of the LigN protein, wild-type and three mutant forms of the LigN protein were separately expressed in recombinant form in E.coli and purified to apparent homogeneity by immobilised metal ion affinity chromatography (IMAC). Non-isotopic DNA ligase activity assays using lambda DNA restriction fragments with 12 bp cos cohesive ends were used to show that LigN activity was dependent on addition of divalent cations and salt. No activity was detected in the absence of KCl, whereas maximum activity could be detected at 3.2 M KCl, close to the intracellular KCl concentration of Hfx. volcanii cells. CONCLUSION: LigN is unique amongst characterised DNA ligase enzymes in displaying maximal DNA strand joining activity at high (> 3 M) salt levels. As such the LigN enzyme has potential both as a novel tool for biotechnology and as a model enzyme for studying the adaptation of proteins to high intracellular salt levels. PMID- 17132165 TI - Correcting ligands, metabolites, and pathways. AB - BACKGROUND: A wide range of research areas in bioinformatics, molecular biology and medicinal chemistry require precise chemical structure information about molecules and reactions, e.g. drug design, ligand docking, metabolic network reconstruction, and systems biology. Most available databases, however, treat chemical structures more as illustrations than as a datafield in its own right. Lack of chemical accuracy impedes progress in the areas mentioned above. We present a database of metabolites called BioMeta that augments the existing pathway databases by explicitly assessing the validity, correctness, and completeness of chemical structure and reaction information. DESCRIPTION: The main bulk of the data in BioMeta were obtained from the KEGG Ligand database. We developed a tool for chemical structure validation which assesses the chemical validity and stereochemical completeness of a molecule description. The validation tool was used to examine the compounds in BioMeta, showing that a relatively small number of compounds had an incorrect constitution (connectivity only, not considering stereochemistry) and that a considerable number (about one third) had incomplete or even incorrect stereochemistry. We made a large effort to correct the errors and to complete the structural descriptions. A total of 1468 structures were corrected and/or completed. We also established the reaction balance of the reactions in BioMeta and corrected 55% of the unbalanced (stoichiometrically incorrect) reactions in an automatic procedure. The BioMeta database was implemented in PostgreSQL and provided with a web-based interface. CONCLUSION: We demonstrate that the validation of metabolite structures and reactions is a feasible and worthwhile undertaking, and that the validation results can be used to trigger corrections and improvements to BioMeta, our metabolite database. BioMeta provides some tools for rational drug design, reaction searches, and visualization. It is freely available at http://www.cmbi.ru.nl/biometa/ provided that the copyright notice of all original data is cited. The database will be useful for querying and browsing biochemical pathways, and to obtain reference information for identifying compounds. However, these applications require that the underlying data be correct, and that is the focus of BioMeta. PMID- 17132166 TI - High yield recombinant penicillin G amidase production and export into the growth medium using Bacillus megaterium. AB - BACKGROUND: During the last years B. megaterium was continuously developed as production host for the secretion of proteins into the growth medium. Here, recombinant production and export of B. megaterium ATCC14945 penicillin G amidase (PGA) which is used in the reverse synthesis of beta-lactam antibiotics were systematically improved. RESULTS: For this purpose, the PGA leader peptide was replaced by the B. megaterium LipA counterpart. A production strain deficient in the extracellular protease NprM and in xylose utilization to prevent gene inducer deprivation was constructed and employed. A buffered mineral medium containing calcium ions and defined amino acid supplements for optimal PGA production was developed in microscale cultivations and scaled up to a 2 Liter bioreactor. Productivities of up to 40 mg PGA per L growth medium were reached. CONCLUSION: The combination of genetic and medium optimization led to an overall 7-fold improvement of PGA production and export in B. megaterium. The exclusion of certain amino acids from the minimal medium led for the first time to higher volumetric PGA activities than obtained for complex medium cultivations. PMID- 17132167 TI - Salivary flow rate, pH, and concentrations of calcium, phosphate, and sIgA in Brazilian pregnant and non-pregnant women. AB - BACKGROUND: Studies on salivary variables and pregnancy in Latin America are scarce. This study aimed to compare salivary flow rate, pH, and concentrations of calcium, phosphate, and sIgA of unstimulated whole saliva in pregnant and non pregnant Brazilians. METHODS: Cross-sectional study. Sample was composed by 22 pregnant and 22 non-pregnant women attending the Obstetrics and Gynecology Clinics, Sao Lucas Hospital, in Porto Alegre city, South region of Brazil. Unstimulated whole saliva was collected to determine salivary flow rate, pH, and biochemical composition. Data were analyzed by Student t test and ANCOVA (two tailed alpha = 0.05). RESULTS: No difference was found for salivary flow rates and concentrations of total calcium and phosphate between pregnant and non pregnant women (p > 0.05). Pregnant women had lower pH (6.7) than non-pregnant women (7.5) (p < 0.001), but higher sIgA level (118.9 mg/L) than the latter (90.1 mg/L) (p = 0.026). CONCLUSION: Some of the tested variables of unstimulated whole saliva were different between pregnant and non-pregnant Brazilians in this sample. Overall, the values of the tested salivary parameters were within the range of international references of normality. PMID- 17132168 TI - Longitudinal changes in HIV-specific IFN-gamma secretion in subjects who received Remune vaccination prior to treatment interruption. AB - BACKGROUND: Despite the benefits of highly active antiretroviral therapy (HAART) for suppressing viral replication in HIV infection, virus persists and rebounds during treatment interruption (TI). This study explored whether HAART intensification with Remune vaccination before TI can boost HIV-1-specific immunity, leading to improved control of viremia off HAART. METHODS: Ten chronically HIV-infected adults were enrolled in this proof of concept study. After a 6-month HAART intensification phase with didanosine, hydroxyurea, granulocyte-macrophage colony-stimulating factor, (GM-CSF), and a first dose of Remune (HIV-1 Immunogen), HAART was discontinued. Patients continued to receive Remune every 3 months until the end of study. HAART was restarted if viral load did not fall below 50,000 copies/ml of plasma within 3 months or if CD4+ counts decreased to <200 cells/mm3. HIV-specific immunity was monitored with the interferon-gamma (IFN-gamma) ELISPOT assay. RESULTS: All subjects experienced viral rebound during TIs. Although the magnitude and breadth of HIV-specific responses to HLA-restricted optimal peptide panels and Gag p55 peptide pools increased and viral load decreased by 0.44 log10 units from TI#1 to TI#2, no significant correlations between these parameters were observed. The patients spent 50.4% of their 36 months follow up off HAART. CONCLUSION: Stopping HAART in this vaccinated population induced immune responses that persisted after therapy was restarted. Induction of HIV-specific immunity beyond IFN-gamma secretion may be contributing to better control of viremia during subsequent TIs allowing for long periods off HAART. PMID- 17132169 TI - Gender plays no role in student ability to perform on computer-based examinations. AB - BACKGROUND: To see if there is a difference in performance when students switch from traditional paper-and-pencil examinations to computer-based examinations, and to determine whether there are gender differences in student performance in these two examination formats. METHODS: This study involved first year medical students at the University of Illinois at Urbana-Champaign over three Academic Years 2002-03/2003-04 and 2003-05. Comparisons of student performance by overall class and gender were made. Specific comparisons within courses that utilized both the paper-and-pencil and computer formats were analyzed. RESULTS: Overall performance scores for students among the various Academic Years revealed no differences between exams given in the traditional pen-and-paper and computer formats. Further, when we looked specifically for gender differences in performance between these two testing formats, we found none. CONCLUSION: The format for examinations in the courses analyzed does not affect student performance. We find no evidence for gender differences in performance on exams on pen-and-paper or computer-based exams. PMID- 17132170 TI - Post-exposure prophylaxis for SIV revisited: animal model for HIV prevention. AB - BACKGROUND: A 4-week, uninterrupted treatment with 9-(2-phosphonyl methoxypropyly)adenine (PMPA, commonly called tenofovir) completely prevents simian immunodeficiency virus (SIVmne) infection in cynomolgus macaques if treatment begins within 24 hours after SIVmne inoculation, but is less effective if treatment is delayed or duration of treatment is shortened. Critical factors for efficacy include timing and duration of treatment, potency of antiretroviral drug and a contribution from antiviral immune responses. Therefore, we evaluated the impact of one or more treatment interruptions plus SIVmne re-exposures on efficacy of PMPA treatment to prevent SIVmne infection in cynomolgus macaques. We also evaluated whether macaques with pre-existing SIV immune responses show increased efficacy of treatment. Eight PMPA-treated, virus-negative and seronegative macaques, and five PMPA-treated, virus-negative but weakly or strongly seropositive macaques were re-inoculated with SIVmne and treated with PMPA starting 24 hr post inoculation. Thereafter, they received either a 5-week treatment involving one interruption plus one SIVmne challenge or a 10-week treatment involving six interruptions plus six SIVmne challenges early during treatment. Parameters measured were plasma SIV RNA, SIV-antibody response, CD4+ T lymphocyte subsets and in vivo CD8+ cell-suppression of virus infection. RESULTS: All seronegative macaques developed persistent antibody response beginning 4 to 8 weeks after stopping PMPA-treatment in absence of viremia in a majority of macaques and coinciding with onset of intermittent viremia in other macaques. In contrast, all weakly or strongly seropositive macaques showed immediate increase in titers (> 1600) of SIV antibodies, even before the end of PMPA-treatment, and in absence of detectable viremia. However, in vivo CD8+-cell depletion revealed CD8 cell-suppression of viremia and persistence of virus in the macaques as long as 2 years after PMPA-treatment, even in aviremic macaques. Unlike untreated macaques, a treated macaque controlled viral replication and blocked CD4+ T cell depletion when challenged with a heterologous chimeric SIV/HIV-1 virus called SHIV89.6P. CONCLUSION: A single interruption plus one SIVmne challenge was as sufficient as six interruptions plus six SIVmne challenges in reducing efficacy of PMPA, but results in long-term persistence of virus infection suppressed by CD8+ cells. Efficacy of PMPA treatment was highest in macaques with pre-existing SIV immune responses. PMID- 17132171 TI - Quantitative phase imaging with scanning holographic microscopy: an experimental assessment. AB - This paper demonstrates experimentally how quantitative phase information can be obtained in scanning holographic microscopy. Scanning holography can operate in both coherent and incoherent modes, simultaneously if desired, with different detector geometries. A spatially integrating detector provides an incoherent hologram of the object's intensity distribution (absorption and/or fluorescence, for example), while a point detector in a conjugate plane of the pupil provides a coherent hologram of the object's complex amplitude, from which a quantitative measure of its phase distribution can be extracted. The possibility of capturing simultaneously holograms of three-dimensional specimens, leading to three dimensional reconstructions with absorption contrast, reflectance contrast, fluorescence contrast, as was previously demonstrated, and quantitative phase contrast, as shown here for the first time, opens up new avenues for multimodal imaging in biological studies. PMID- 17132172 TI - Work factors and psychological distress in nurses' aides: a prospective cohort study. AB - BACKGROUND: Nurses' aides (assistant nurses), the main providers of practical patient care in many countries, are doing both emotional and heavy physical work, and are exposed to frequent social encounters in their job. There is scarce knowledge, though, of how working conditions are related to psychological distress in this occupational group. The aim of this study was to identify work factors that predict the level of psychological distress in nurses' aides. METHODS: The sample of this prospective study comprised 5076 Norwegian nurses' aides, not on leave when they completed a mailed questionnaire in 1999. Of these, 4076 (80.3 %) completed a second questionnaire 15 months later. A wide spectrum of physical, psychological, social, and organisational work factors were measured at baseline. Psychological distress (anxiety and depression) was assessed at baseline and follow-up by the SCL-5, a short version of Hopkins Symptom Checklist 25. RESULTS: In a linear regression model of the level of psychological distress at follow-up, with baseline level of psychological distress, work factors, and background factors as independent variables, work factors explained 2 % and baseline psychological distress explained 34 % of the variance. Exposures to role conflicts, exposures to threats and violence, working in apartment units for the aged, and changes in the work situation between baseline and follow-up that were reported to result in less support and encouragement were positively associated with the level of psychological distress. Working in psychiatric departments, and changes in the work situation between baseline and follow-up that gave lower work pace were negatively associated with psychological distress. CONCLUSION: The study suggests that work factors explain only a modest part of the psychological distress in nurses' aides. Exposures to role conflicts and threats and violence at work may contribute to psychological distress in nurses' aides. It is important that protective measures against violent patients are implemented, and that occupational health officers offer victims of violence appropriate support or therapy. It is also important that health service organisations focus on reducing role conflicts, and that leaders listen to and consider the views of the staff. PMID- 17132173 TI - Diagnostic value of blood inflammatory markers for detection of acute appendicitis in children. AB - BACKGROUND: Acute appendicitis (AA) is a common surgical problem that is associated with an acute-phase reaction. Previous studies have shown that cytokines and acute-phase proteins are activated and may serve as indicators for the severity of appendicitis. The aim of this study was to compare diagnostic value of different serum inflammatory markers in detection of phlegmonous or perforated appendicitis in children. METHODS: Data were collected prospectively on 211 consecutive children. Laparotomy was performed for suspected AA for 189 patients. Patients were subdivided into groups: nonsurgical abdominal pain, early appendicitis, phlegmonous or gangrenous appendicitis, perforated appendicitis. White blood cell count (WBC), serum C-reactive protein (CRP), interleukin-6 (IL 6), tumor necrosis factor alpha (TNF-alpha), acid alpha1-glycoprotein (alpha1GP), endotoxin, and erythrocyte sedimentation reaction (ESR) were estimated ad the time of admission. The diagnostic performance was analyzed using receiver operating characteristic (ROC) curves. RESULTS: WBC count, CRP and IL-6 correlated significantly with the severity of appendiceal inflammation. Identification of children with severe appendicitis was supported by IL-6 or CRP but not WBC. Between IL-6 and CRP, there were no significant differences in diagnostic use. CONCLUSION: Laboratory results should be considered to be integrated within the clinical assessment. If used critically, CRP and IL-6 equally provide surgeons with complementary information in discerning the necessity for urgent operation. PMID- 17132174 TI - Post traumatic intra thoracic spleen presenting with upper GI bleed!--a case report. AB - BACKGROUND: Isolated splenic vein thrombosis with left sided portal hypertension is a rare cause of upper gastrointestinal bleed. Diagnosis is difficult and requires a high index of suspicion, especially in patients presenting with gastrointestinal bleed in the presence of splenomegaly and normal liver function tests. CASE PRESENTATION: A 64 year old male presented with haematemesis and melaena. An upper gastrointestinal endoscopy revealed the presence of antral erosions in the stomach and fundal varices. A computerised tomography scan of abdomen confirmed the presence of a diaphragmatic tear and the spleen to be lying in the left hemi thorax. The appearances of the splenic vein on the scan were consistent with thrombosis. CONCLUSION: Left sided portal hypertension as a result of isolated splenic vein thrombosis secondary to trauma is rare. The unusual presentation of our case, splenic herniation into the left hemithorax, causing fundal varices leading to upper gastrointestinal bleed 28 years after the penetrating injury, makes this case most interesting. We believe that this has not been reported in literature before. PMID- 17132175 TI - Inland hypersaline lakes and the brine shrimp Artemia as simple models for biodiversity analysis at the population level. AB - Biodiversity can be measured at different hierarchical levels, from genetic diversity within species to diversity of ecosystems, though policy-makers tend to use species richness. The 2010 goal of reducing biodiversity loss, agreed by the subscribers to the Convention on Biological Diversity, requires simple and reliable protocols to evaluate biodiversity at any level in a given ecosystem. Stakeholders, particularly policy makers, need to understand how ecosystem components interact to produce social and economic benefits on the long run, whilst scientists are expected to fulfil this demand by testing and modelling ideally simple (low diversity) ecosystems, and by monitoring key species. This work emphasizes the unique opportunity offered by inland, isolated salt lakes and the brine shrimp Artemia, an example of biodiversity contained at the intra specific level, as simple models to understand and monitor biodiversity, as well as to assess its predicted positive association with ecosystem stability. In addition to having well identified species and strains and even clones, that allow to test reproductive effects (sexual versus asexual), Artemia benefits from the possibility to set up experimental testing at both laboratory scale and outdoor pond systems, for which a comprehensive cyst bank with sufficient amount of samples from all over the world is available. PMID- 17132176 TI - Haloperidol treatment induces tissue- and sex-specific changes in DNA methylation: a control study using rats. AB - BACKGROUND: We previously found that there is a subtle difference in the global methylation state of blood leukocyte DNA between male subjects with and without schizophrenia. The aim of the current study was to determine whether this difference was a primary effect of the disease state, or a secondary effect of antipsychotics administered to these patients. METHODS: We examined the methyl cytosine (mC) content of DNA from the leukocytes, brain, and liver of rats using high performance liquid chromatography. A total of 40 male and female rats received for 21 days daily injection of haloperidol or vehicle solution alone. RESULTS: In control rats injected with buffer only, there was a sex-dependent difference in mC content in leukocyte DNA (male > female; P = 0.028, n = 10), similar to our previous observations in human peripheral leukocytes. No difference in mC content between the sexes was observed in the brain or liver in buffer-treated animals. Haloperidol treatment slightly decreased the mC content of leukocytes in male rats, but unexpectedly, increased the mC content of leukocytes in females. We observed a trend toward a higher level of mC in the liver in both sexes following haloperidol treatment, compared to buffer-treated animals. In contrast, haloperidol treatment resulted in a decrease in mC content in the brain in females, and this difference was statistically significant (P = 0.026). CONCLUSION: These results indicate that haloperidol can affect DNA methylation states in the brain, as well as in certain other tissues, and raise the possibility that antipsychotic drugs play a role in the observed disparity in mC content in male subjects with and without schizophrenia. PMID- 17132177 TI - Optimization of extracranial stereotactic radiation therapy of small lung lesions using accurate dose calculation algorithms. AB - BACKGROUND: The aim of this study was to compare and to validate different dose calculation algorithms for the use in radiation therapy of small lung lesions and to optimize the treatment planning using accurate dose calculation algorithms. METHODS: A 9-field conformal treatment plan was generated on an inhomogeneous phantom with lung mimics and a soft tissue equivalent insert, mimicking a lung tumor. The dose distribution was calculated with the Pencil Beam and Collapsed Cone algorithms implemented in Masterplan (Nucletron) and the Monte Carlo system XVMC and validated using Gafchromic EBT films. Differences in dose distribution were evaluated. The plans were then optimized by adding segments to the outer shell of the target in order to increase the dose near the interface to the lung. RESULTS: The Pencil Beam algorithm overestimated the dose by up to 15% compared to the measurements. Collapsed Cone and Monte Carlo predicted the dose more accurately with a maximum difference of -8% and -3% respectively compared to the film. Plan optimization by adding small segments to the peripheral parts of the target, creating a 2-step fluence modulation, allowed to increase target coverage and homogeneity as compared to the uncorrected 9 field plan. CONCLUSION: The use of forward 2-step fluence modulation in radiotherapy of small lung lesions allows the improvement of tumor coverage and dose homogeneity as compared to non modulated treatment plans and may thus help to increase the local tumor control probability. While the Collapsed Cone algorithm is closer to measurements than the Pencil Beam algorithm, both algorithms are limited at tissue/lung interfaces, leaving Monte-Carlo the most accurate algorithm for dose prediction. PMID- 17132178 TI - What makes us human? A biased view from the perspective of comparative embryology and mouse genetics. AB - For a neurobiologist, the core of human nature is the human cerebral cortex, especially the prefrontal areas, and the question "what makes us human?" translates into studies of the development and evolution of the human cerebral cortex, a clear oversimplification. In this comment, after pointing out this oversimplification, I would like to show that it is impossible to understand our cerebral cortex if we focus too narrowly on it. Like other organs, our cortex evolved from that in stem amniotes, and it still bears marks of that ancestry. More comparative studies of brain development are clearly needed if we want to understand our brain in its historical context. Similarly, comparative genomics is a superb tool to help us understand evolution, but again, studies should not be limited to mammals or to comparisons between human and chimpanzee, and more resources should be invested in investigation of many vertebrate phyla. Finally, the most widely used rodent models for studies of cortical development are of obvious interest but they cannot be considered models of a "stem cortex" from which the human type evolved. It remains of paramount importance to study cortical development directly in other species, particularly in primate models, and, whenever ethically justifiable, in human. PMID- 17132179 TI - Transcranial magnetic stimulation: a stimulating new method for treating depression, but saddled with the same old problems. PMID- 17132180 TI - Healing bodies, saving souls. Medical missions in Asia and Africa. PMID- 17132197 TI - Organizational assessment in paediatric primary care: development and initial validation of the primary care organizational questionnaire. AB - Primary care in the United States is undergoing many changes. Reliable and valid instruments are needed to assess the effects of these changes. The Primary Care Organizational Questionnaire (PCOQ), a 56-item 5-point Likert scale survey that evaluates interactions among members of the clinic/practice and job-related attributes, was administered to clinicians and staff in 36 primary care practices serving paediatric populations in Connecticut. A priori scales were reliable (Cronbach alpha > or = 0.7). Analysis of variance (ANOVA) showed greater heterogeneity across clinics than within clinics for 13 of 15 a priori scales, which were then included in a principal component factor analysis with varimax rotation. Eigenvalue analysis showed nine significant factors, largely similar to the a priori scales, indicating concurrent construct validity. Further research will ascertain the utility of the PCOQ in predicting the effectiveness of primary care practices in implementing disease management programmes. PMID- 17132198 TI - Patient reciprocity and physician burnout: what do patients bring to the patient physician relationship? AB - Despite its criticality to the provision of health care, little is known about how the patient-physician relationship influences burnout. This article seeks to understand how patient performance (e.g. being informative about needs) during office visits is associated with perceived reciprocity in the patient-physician relationship, which is in turn associated with physician burnout. To that end, we report the results of a cross-sectional survey of 252 matched pairs of patients and their primary care physicians about a recent office visit. The findings support a social exchange model of burnout that suggests that patient stressors and patient performance predict perceived reciprocity and subsequent burnout. Interestingly, patients' perceptions of their performance differed from physicians' perceptions; physicians' perceptions of performance fit the social exchange model better than patients' perceptions of performance.The present work suggests that while they are a source of demands, patients also provide resources that are critical to the patient-physician relationship. To the extent that we can encourage these resources, we can improve perceived reciprocity and reduce burnout in physicians. PMID- 17132199 TI - Proposed hub-network model for tissue procurement: the case of Arizona. AB - A single institution is not well positioned to manage the process of tissue donations to maximize utility and minimize the costs associated with such transactions. The proposed creation of a 'hub' organization to service the needs and demands associated with tissue procurement and utilization is outlined. The case of Arizona is used as a case study for the proposed model building on existing network in tissue donation and research. The proposed model would consequently be viewed as legitimate by various network stakeholders, including the general public. Finally, the proposed hub-network would be better able to manage the donation, procurement, storage, preservation, distribution, and use of human tissues for all of the potential uses in a single state. PMID- 17132200 TI - Consumer determinants of the use of health plan information in plan selection. AB - One of the major issues in achieving optimum levels of performance in health-care markets is to enhance consumer understanding of their health plan choices in order to facilitate the expansion of 'high-value' health plans at the expense of 'low-value' health plans. The Federal government offers employees many choices of health plans and provides large amounts of information on all of these options through (1) comparative written health plan information, (2) information from the health plans themselves, and (3) comparative health plan information on the Internet. The present study examines the degree to which 1722 Federal employees in the Department of Health and Human Services utilized health plan information from the above three sources in making their annual health plan selection. Results indicate that most employees (64%) used at least one information source, with written information from health plans the most common (53%), followed by comparative written information in The Guide (32%) and the Internet (16%). Those employees who regularly search for information prior to making an important purchase, those with a short time in their current plan, those with family coverage, Whites, African-Americans, and men were all more likely to use health plan information to make their annual choice. The Internet was accessed more often by younger and higher paid employees. Implications for policy and future research are discussed. PMID- 17132201 TI - An analysis of the strategies for public hospitals. AB - Today, more than ever, the variables that define the health-care environment (demand, costs, system deregulation) are undergoing rapid change, thus obliging hospital administrators to develop and implement competitive strategies in order to survive in the increasingly competitive environment of hospital management. The primary objective of this paper is to identify strategies in a sample of Spanish public hospitals. Following this, we try to detect groups of organizations using similar strategies and ultimately analyse their influence on hospital performance. PMID- 17132202 TI - The utilization of systematic outcome mapping to improve performance management in health care. AB - Performance management is an important mechanism for ensuring accountability and improving the quality of health-care services. The last decade has witnessed a proliferation in the development of performance measurement systems for assessing health-care processes and outcomes at the program, hospital, district, system and national level. This has allowed for comparison and benchmarking between similar aspects of care at each of these levels. Unfortunately, most performance systems are devoid of clear mechanisms for translating feedback from measures into strategies for action, thus leaving largely unfulfilled the quality and management aspect necessary to improve health-care services. Therefore, the thinking that goes into designing these systems must change. This article outlines a management framework called systematic outcome mapping that provides for performance management rather than just performance measurement by allowing for quality improvement to be built into performance indicator development. It utilizes evidence-based medicine and expert consensus opinion to establish linkages between processes of care and their outcomes with the clear intent that feedback from information provided by performance indicators can be used to modify health-care activities so as to improve health outcomes. This fulfils the quality improvement aspect of performance measurement and makes it an integral part of a performance management framework that reinforces organizational learning through feedback from outcomes and the assessment of organizational routines. PMID- 17132203 TI - NSAID injury to the gastrointestinal tract: evidence that NSAIDs interact with phospholipids to weaken the hydrophobic surface barrier and induce the formation of unstable pores in membranes. AB - In this review, we have discussed our current understanding of the barrier properties that are in place to protect the upper gastrointestinal mucosa from luminal acid, and the pathogenic mechanism by which nonsteroidal anti inflammatory drugs (NSAIDs) induce injury to the gastrointestinal tract. The changes in our view of the importance of NSAID-induced cyclo-oxygenase (COX) inhibition on the pathogenesis and prevention of NSAID-induced gastrointestinal injury is presented. The focus of this paper has been placed on the effects of NSAIDs on the mucosal surface, and specifically the effect of these powerful drugs in inducing changes in the hydrophobicity, fluidity, biomechanical and permeability properties of extracellular and membrane phospholipids. Lastly, recent evidence is presented that salicylic acid and related NSAIDs may alter the stability of membranes, inducing the formation of unstable pores that may lead to back-diffusion of luminal acid and membrane rupture. This understanding of the interaction of NSAIDs with membrane phospholipids may prove valuable in the design of novel NSAID formulations with reduced gastrointestinal side-effects. PMID- 17132204 TI - Formulation and evaluation of nimodipine-loaded lipid microspheres. AB - The purpose of this study was to develop an alternative, improved and better tolerated formulation and investigate the pharmacokinetic profile of the new formulation of nimodipine (NM) compared with nimodipine ethanol solutions. Lipid microspheres (LMs) prepared using lecithin and vegetable oils have attracted a lot of interest owing to their versatile properties, such as non-immunogenicity, being easily biodegradable and exhibiting high entrapment efficiency. NM incorporated in LMs could reduce irritation by avoiding the use of ethanol as a solubilizer. The solubility of NM was also increased by dissolving it in the oil phase. The particle size distribution, zeta potential, entrapment efficacy and assay of the NM-loaded LMs were found to be 188.2+/-5.4 nm, -31.6 mV, 94.2% and 1.04 mg mL(-1), respectively. The preparation was stable for 1 year at 4-10 degrees C. The formulation and some physicochemical properties of NM-loaded LMs were investigated. The pharmacokinetic and biodistribution studies were performed in rats at a dose of 1.2 mg kg(-1). From the observed data, there is no obvious retention of NM-loaded LMs in plasma. Moreover, incorporation of NM in LMs did not alter the tissue distribution significantly except for the relatively greater drug accumulation in the liver and spleen. The stimulation studies demonstrate that LMs of NM reduce irritation markedly compared with NM solutions. These results suggest that the LM system is a promising option to replace NM ethanol solutions as an intravenous treatment. PMID- 17132205 TI - Improvement of the agitation granulation method to prepare granules containing a high content of a very hygroscopic drug. AB - This study describes a new approach to the preparation of a granulate with a high content of a very hygroscopic powder or drug, using the agitation granulation method, and the development of a tablet formulation using these granulates. A Chinese medicine extract, Hatimi-zio-gan, was used as the model of a very hygroscopic drug. Among the several excipients tested, only porous calcium silicate could be used to prepare granules, with a mixing ratio (extract to porous calcium silicate) from 2:1 to 20:1. With other excipients, very large lumps were formed during the granulation process. The best mixing ratio of extract to porous calcium silicate was 6:1. For preparation of the granules, water could be added to the mixed powder within a range of 1- to 4-times the amount of porous calcium silicate. From these results, it was concluded that the ability of porous calcium silicate to hold large amounts of water in its numerous pores may allow for the preparation of granulates with a high content of very hygroscopic drugs. Starch with partial alpha-links, carboxymethyl starch sodium salt and crospovidone were used for selection of the disintegration agent. When crospovidone was used as a disintegration agent, tablets containing about 70% of the Chinese medicine extract disintegrated in less than 7 min, with good dissolution rates. The same process was applied to extracts of Hotyu-ekki-to, Syo seiryu-to, Boi-ogi-to and Bohu-tusyo-san. The absorption of paeoniflorin, a characteristic monoterpene glucoside contained in Hatimi-zio-gan extract, was evaluated in beagle dogs after oral administration of the Hatimi-zio-gan tablets prepared in this study. The values of C(max) and AUC obtained after administration of the tablets prepared in this study were significantly greater than those obtained for commercial tablets. PMID- 17132206 TI - Pharmacokinetics of selected stilbenes: rhapontigenin, piceatannol and pinosylvin in rats. AB - The pharmacokinetics of piceatannol, pinosylvin and rhapontigenin were characterized in male Sprague-Dawley rats after single intravenous doses of 10 mg kg(-1) of each stilbene. Serial blood samples were collected via a catheter inserted into the right jugular vein and plasma samples were analysed for the selected stilbenes concentrations using reverse phase HPLC methods. After an acute intravenous dose of piceatannol, plasma AUC, urine t(1/2), CL and V(d) were 8.48+/-2.48 micro g h mL(-1), 19.88+/-5.66 h, 2.13+/-0.92 Lh(-1) kg(-1) and 10.76+/-2.88 L kg(-1)(mean+/-s.e.m.), respectively. The acute intravenous dose of pinosylvin yielded the plasma AUC, urine t(1/2), CL and V(d) values of 5.23+/ 1.20 micro g h mL(-1), 13.13+/-2.05 h, 1.84+/-0.44 Lh(-1) kg(-1) and 2.29+/-0.56 L kg(-1)(mean+/-s.e.m.), respectively. Rhapontigenin intravenous dosing yielded the plasma AUC, urine t(1/2), CL and V(d) values of 8.39+/-0.10 micro g h mL(-1), 25.31+/-1.46 h, 1.18+/-0.035 Lh(-1) kg(-1) and 11.05+/-0.17 L kg(-1)(mean+/ s.e.m.), respectively. Each stilbene was extensively glucuronidated. These stilbenes were predominantly eliminated via non-urinary routes. All three stilbenes were highly distributed into tissues and were highly extracted by the liver. The detectable plasma half-lives of these xenobiotics appear to be relatively short. However, utilizing urinary concentration-time data, much longer elimination half-lives were evident. The estimates of oral bioavailability characterize these stilbenes as poorly bioavailable compounds. PMID- 17132207 TI - Distribution of lovastatin to bone and its effect on bone turnover in rats. AB - Statins, the widely used lipid-lowering drugs, are inhibitors of 3-hydroxy-3 methyl-glutaryl coenzyme A (HMG-CoA) reductase, which catalyses a rate-limiting step in the biosynthesis of cholesterol. Many previous reports show that statins can act both as bone anabolic and as anti-resorptive agents but their beneficial effects on bone turnover are still controversial. Considering their high liver specificity and low oral bioavailability, the distribution of statins to the bone microenvironment is questionable. In this study, the distribution of lovastatin and its active metabolites to bone, with respect to plasma and liver compartments, was examined after oral and intravenous administration in female rats. As compared with oral administration, the distribution of lovastatin to the bone compartment was significantly enhanced after intravenous administration. Further, the effect of lovastatin on bone turnover was studied in-vitro and in vivo to assess its anti-osteoporotic potential. Lovastatin acid but not lovastatin was found to inhibit parathyroid-hormone-induced bone resorption in an in-vitro chick embryo bone assay. Oral, as well as intravenous, short-term lovastatin treatment significantly reduced the serum total cholesterol, serum total alkaline phosphatase and urinary crosslinks in ovariectomized rats. In accordance with its increased distribution to the bone compartment, intravenously administered lovastatin was more effective in reducing the ovariectomy-induced increase in markers of bone metabolism, especially urinary crosslinks. The findings of this study suggest that statins inhibit bone resorption and that their anti-resorptive efficacy can be increased by administering them by routes other than oral so as to achieve their enhanced concentration in bone. PMID- 17132208 TI - Permeation of WIN 55,212-2, a potent cannabinoid receptor agonist, across human tracheo-bronchial tissue in vitro and rat nasal epithelium in vivo. AB - The aim of this study was to investigate the intranasal absorption of R-(+)-WIN 55,212-2 mesylate in vivo and in vitro. Permeation experiments of R-(+)-WIN 55,212-2 formulations with 2% dimethyl-beta-cyclodextrin (DMbetaCD), 2% trimethyl beta-cyclodextrin (TMbetaCD) or 2% randomly methylated-beta-cyclodextrin (RAMbetaCD) in 1:1 propylene glycol/saline and 1.5% propylene glycol +3% Tween 80 in saline were conducted using EpiAirway tissue and an anesthetized rat nasal absorption model, respectively. Samples were analysed by liquid chromatography mass spectrometry. Mucosal tolerance was screened using paracellular marker permeation and tissue viability as indices. Nasal absorption of WIN 55,212-2 was rapid, with a t(max) (time of peak concentration) of 0.17 to 0.35 h in vivo. Relative to 1.5% propylene glycol +3% Tween 80 (control), 1:1 propylene glycol/saline, RAMbetaCD, DMbetaCD and TMbetaCD resulted in 24-, 20-, 17- and 10 fold WIN 55,212-2 permeation increases in vitro, respectively. The in vivo absolute bioavailabilities were also increased with 1:1 propylene glycol/saline, RAMbetaCD, DMbetaCD and TMbetaCD compared to 1.5% propylene glycol +3% Tween 80 (0.15 vs. 0.66-0.77). The viability of the EpiAirway tissues was significantly reduced by DMbetaCD and TMbetaCD formulations. This study showed that WIN 55,212 2 mesylate can be delivered via the nasal route. Absorption of R-(+)-WIN 55,212-2 was rapid and bioavailability was significantly improved using methylated cyclodextrins and propylene glycol-based cosolvent. PMID- 17132209 TI - In-vitro study on the competitive binding of diflunisal and uraemic toxins to serum albumin and human plasma using a potentiometric ion-probe technique. AB - The competitive binding of diflunisal and three well-known uraemic toxins (3 indoxyl sulfate, indole-3-acetic acid and hippuric acid) to bovine serum albumin (BSA), human serum albumin (HSA) and human plasma was studied by direct potentiometry. The method used the potentiometric drug ion-probe technique with a home-made ion sensor (electrode) selective to the drug anion. The site-oriented Scatchard model was used to describe the binding of diflunisal to BSA, HSA and human plasma, while the general competitive binding model was used to calculate the binding parameters of the three uraemic toxins to BSA. Diflunisal binding parameters, number of binding sites, n(i) and association constants for each class of binding site, K(i), were calculated in the absence and presence of uraemic toxins. Although diflunisal exhibits high binding affinity for site I of HSA and the three uraemic toxins bind primarily to site II, strong interaction was observed between the drug and the three toxins, which were found to affect the binding of diflunisal on its primary class of binding sites on both BSA and HSA molecules and on human plasma. These results are strong evidence that the decreased binding of diflunisal that occurs in uraemic plasma may not be solely attributed to the lower albumin concentration observed in many patients with renal failure. The uraemic toxins that accumulate in uraemic plasma may displace the drug from its specific binding sites on plasma proteins, resulting in increased free drug plasma concentration in uraemic patients. PMID- 17132210 TI - Classification of torasemide based on the Biopharmaceutics Classification System and evaluation of the FDA biowaiver provision for generic products of CLASS I drugs. AB - The biopharmaceutical properties of an in-house developed new crystal modification of torasemide (Torasemide N) were investigated in comparison with the most well known crystal modification form of torasemide (Torasemide I) in order to classify the drug according to the Biopharmaceutics Classification System (BCS), and to evaluate the data in line with current US Food and Drug Administration (FDA) guidance (with biowaiver provision for Class I drugs) to determine if the biowaiver provision could be improved. The solubility profiles of Torasemide I and Torasemide N were determined, and tablets prepared from both forms of the drug were studied for in-vitro release characteristics in media recommended by the current FDA guidance for biowaiver of generic products, and in other media considered more appropriate for the purpose than the ones recommended by the FDA. Two separate bioequivalence studies in healthy humans (following oral administration) were performed with two test products (both prepared from Torasemide I) against a single reference product (prepared from Torasemide N). The absorption profiles of the drug from the tablets were determined by deconvolution for comparison with the in-vitro release profiles to determine the appropriateness of some dissolution media for predicting in-vivo performance and to determine the comparative rate and extent of absorption. The drug was absorbed from the tested products quickly and almost completely (about 95% within 3.5 h of administration). However, one test product failed to meet the bioequivalence criteria and had a significant initial lower absorption rate profile compared with the reference product (P< or =0.05), whereas the other product was bioequivalent and had a similar absorption profile to the reference product. A dissolution medium at pH 5.0, in which torasemide has minimum solubility, was found to be more discriminatory than the media recommended by the FDA. Torasemide has been classified as a Class I drug according to the BCS up to a maximum dose of 40 mg and the data suggest that the current FDA guidance could be improved by giving more emphasis to selection of appropriate dissolution media than is given in its current form for approving biowaiver to generic products of Class I drugs. PMID- 17132211 TI - The antioxidant role of pterostilbene in streptozotocin-nicotinamide-induced type 2 diabetes mellitus in Wistar rats. AB - The antioxidant effect of pterostilbene on streptozotocin-nicotinamide-induced diabetic rats has been assessed. The activity of superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase and reduced glutathione was significantly decreased in liver and kidney of diabetic animals when compared with normal control. There were significant improvements in these activities after treatment with pterostilbene at a dose of 40 mg kg(-1) for six weeks. The increased levels of lipid peroxidation measured as thiobarbituric acid reactive substances (TBARS) in liver and kidney of diabetic rats were also normalized by treatment with pterostilbene. Chronic treatment of pterostilbene remarkably reduced the pathological changes observed in liver and kidney of diabetic rats. These results indicated the antioxidant property of pterostilbene. PMID- 17132212 TI - Investigations on the toxicological profile of functionalized fifth-generation poly (propylene imine) dendrimer. AB - Dendrimers have generated tremendous interest in the field of drug delivery. Despite indications of their utility as drug carriers, the inherent cytotoxicity associated with polycationic dendrimers acts as a limiting factor to their clinical applications. Many functionalization strategies have been adopted to mask peripheral amines in order to overcome this limitation. The object of the present investigation was to evaluate the effect of functionalization on the toxicological profile of fifth-generation poly(propylene imine) dendrimer (PPI 5.0G). Four forms of functionalized dendrimers, including protected glycine and phenylalanine, and mannose and lactose functionalized poly(propylene imine) (PPI) dendrimer, were synthesized as prospective drug carriers. These dendrimeric systems were evaluated for haemolytic toxicity, cytotoxicity, immunogenicity and haematological parameters. PPI-5.0G demonstrated a positive charge-based time- and concentration-dependent toxicity profile. Functionalization greatly improved the toxicity profile of the parent dendrimer. Hence it is proposed that these functionalized forms of PPI dendrimer have great potential as bio-compatible drug vehicles. PMID- 17132213 TI - Inhibitory effects of angiotensin II receptor antagonists and leukotriene receptor antagonists on the transport of human organic anion transporter 4. AB - Human organic anion transporter 4 (OAT4) is the only member of the OAT family that is expressed in the placenta and also expressed in kidney. Although OAT4 has been shown to transport certain organic anions as well as other members of the OAT family, fewer numbers of substrates have been identified for OAT4 compared with OAT1 and OAT3, suggesting that the substrate specificity of OAT4 is greater than other OAT members. However, the substrate specificity of OAT4 remains to be investigated in detail. The aim of this study was to examine the effects of various drugs on the OAT4-mediated transport of estrone-3-sulfate, a typical substrate of OAT4, by using human embryonic kidney cells stably transfected with OAT4 (HEK-OAT4). HEK-OAT4 cells exhibited concentration-dependent uptake of estrone-3-sulfate, with a K(m) value of 20.9+/-3.53 microM. Dehydroepiandrosterone sulfate and probenecid potently inhibited estrone-3 sulfate uptake. We also searched for the potential inhibitors of OAT4 and identified candesartan, candesartan cilexetil, losartan, losartan carboxyl (EXP3174) and valsartan as inhibitors of OAT4, with K(i) values of 88.9, 135.2, 24.8, 13.8 and 19.6 microM, respectively. The above angiotensin II receptor antagonists and leukotriene receptor antagonists share a common structural feature, that is the tetrazole group. Although pranlukast is devoid of anionic motifs other than the tetrazole group, it potently inhibited the OAT4-mediated uptake of estrone-3-sulfate, indicating that a tetrazole group may be one important structural feature in substrate recognition by OAT4. PMID- 17132214 TI - Gastroprotective activity and cytotoxic effect of cyperenoic acid derivatives. AB - The gastroprotective effect of the sesquiterpene cyperenoic acid and seven semi synthetic derivatives was assessed in the HCl/ethanol-induced gastric ulcer model in mice. At doses of 25, 50 and 100 mg kg(-1), cyperenoic acid showed a dose dependent gastroprotective effect reducing the lesions by 45 and 75% at 50 and 100 mg kg(-1), respectively. Seven derivatives of the sesquiterpene were prepared and their gastroprotective activity compared at 50 mg kg(-1). The cytotoxicity of the compounds was evaluated in fibroblasts and AGS cells. At 50 mg kg(-1), patchoulan-15-oic acid (compound 8) presented the best gastroprotective effect, reducing the gastric lesions by 86%, with a similar effect to lansoprazole at 20 mg kg(-1). The gastroprotective effect of cyperenol, cyperenoic acid methyl ester and the ethylamide and butylamide from cyperenoic acid were in the same range, reducing the gastric lesions by 72-77%. Cyperenol and cyperenoic acid methyl ester, however, were more cytotoxic with IC50 (concentration that produces a 50% inhibitory effect) values of 44 and 75, 48 and 75 microM against AGS cells and fibroblasts, respectively. The best gastroprotective effect with lower cytotoxicity was found for the compound 8, cyperenoic acid and the p-anisidyl derivative 7. PMID- 17132215 TI - Protective role of gamma-aminobutyric acid against chronic renal failure in rats. AB - The protective effect of gamma-aminobutyric acid (GABA) against chronic renal failure (CRF) was investigated using a remnant kidney model with 5/6 nephrectomized rats. Nephrectomy led to renal dysfunction, which was evaluated via several parameters including serum urea nitrogen, creatinine (Cr) and Cr clearance. However, the administration of GABA ameliorated renal dysfunction, and a longer administration period of GABA increased its protective effect. In addition, nephrectomized control rats showed an elevation in the fractional excretion of sodium (FE(Na)) with an increase in urinary sodium, while GABA led to a significant decline in FE(Na). Moreover, nephrectomy resulted in a decrease of serum albumin and an increase of urinary protein with a change in the urinary protein pattern, whereas the rats administered GABA showed improvement in these changes associated with CRF caused by nephrectomy. This suggests that GABA would inhibit the disease progression and have a protective role against CRF. As one of the risk factors for CRF progression, hypertension was also regulated by GABA. The results also indicate that GABA may play a protective role against CRF through improvement of the serum lipid profile, with reductions in triglyceride and total cholesterol. Furthermore, nephrectomy led to renal oxidative stress with a decrease in the activity of antioxidative enzymes and elevation of lipid peroxidation. The administration of GABA attenuated oxidative stress induced by nephrectomy through an increase in superoxide dismutase and catalase, and decrease in lipid peroxidation. The histopathological lesions, including glomerular, tubular and interstitial lesions, under nephrectomy were also improved by GABA with the inhibition of fibronectin expression. This study demonstrated that GABA attenuated renal dysfunction via regulation of blood pressure and lipid profile, and it also ameliorated the oxidative stress induced by nephrectomy, suggesting the promising potential of GABA in protecting against renal failure progression. PMID- 17132216 TI - D-chiro-inositol found in Cucurbita ficifolia (Cucurbitaceae) fruit extracts plays the hypoglycaemic role in streptozocin-diabetic rats. AB - Cucurbita ficifolia is commonly used as an antihyperglycaemic agent in Asia. However, the mechanism of its action is unknown. Chemically synthesized D-chiro inositol (D-CI), a component of an insulin mediator, has been demonstrated to have antihyperglycaemic effects in rats. In this study, we found that C. ficifolia contained fairly high levels of D-CI, thus, C. ficifolia may be a natural source of D-CI for reducing blood glucose concentrations in diabetics. We evaluated C. ficifolia fruit extract, containing D-CI, for its antihyperglycaemic effect in streptozotocin-induced diabetic rats. Oral administration of C. ficifolia fruit extract containing 10 or 20 mg D-CI kg(-1) body weight for 30 days resulted in significantly lowered levels of blood glucose, and increased levels of hepatic glycogen, total haemoglobin and plasma insulin. An oral glucose tolerance test was performed in fasted diabetic and normal rats, in which there was a significant improvement in blood glucose tolerance in the diabetic rats treated with C. ficifolia fruit extract. The effects were compared with 20 mg kg( 1) body weight chemically synthesized D-CI. Findings from this study demonstrated that C. ficifolia fruit extract was an effective source of D-CI for its hypoglycaemic effects in rats, and therefore may be useful in the treatment of diabetes. PMID- 17132217 TI - Reduction of ciclosporin and tacrolimus nephrotoxicity by plant polyphenols. AB - The immunosuppressants ciclosporin (cyclosporin A, CsA) and tacrolimus can cause severe nephrotoxicity. Since CsA increases free radical formation, this study investigated whether an extract from Camellia sinensis, which contains several polyphenolic free radical scavengers, could prevent nephrotoxicity caused by CsA and tacrolimus. Rats were fed powdered diet containing polyphenolic extract (0 0.1%) starting 3 days before CsA or tacrolimus. Free radicals were trapped with alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone (POBN) and measured using an electron spin resonance spectrometer. Both CsA and tacrolimus decreased glomerular filtration rates (GFR) and caused tubular atrophy, vacuolization and calcification and arteriolar hyalinosis, effects that were blunted by treatment with dietary polyphenols. Moreover, CsA and tacrolimus increased POBN/radical adducts in urine nearly 3.5 fold. Hydroxyl radicals attack dimethyl sulfoxide (DMSO) to produce a methyl radical fragment. Administration of CsA or tacrolimus with (12)C-DMSO produced a 6-line spectrum, while CsA or tacrolimus given with (13)C-DMSO produced a 12-line ESR spectrum, confirming formation of hydroxyl radicals. 4-Hydroxynonenal (4-HNE), a product of lipid peroxidation, accumulated in proximal and distal tubules after CsA or tacrolimus treatment. ESR changes and 4-HNE formation were largely blocked by polyphenols. Taken together, these results demonstrate that both CsA and tacrolimus stimulate free radical production in the kidney, most likely in tubular cells, and that polyphenols minimize nephrotoxicity by scavenging free radicals. PMID- 17132218 TI - In-vitro mutagenic potential and effect on permeability of co-administered drugs across Caco-2 cell monolayers of Rubus idaeus and its fortified fractions. AB - This study investigated the mutagenic, anti-mutagenic and cytotoxic effects of acetone extract of raspberry, Rubus idaeus L. (v. Ottawa) Rosaceae, and the isolated and characterized ellagitannin and anthocyanin fractions thereof, suitable for food applications. The studied raspberry extract and fractions did not show any mutagenic effects determined in the miniaturized Ames test and were not cytotoxic to Caco-2 cells at the used concentrations. However, the anti mutagenic properties were changed (i.e. decreased mutagenicity of 2-nitrofluorene in strain TA98, and slightly increased mutagenicity of 2-aminoanthracene in strain TA100) with metabolic activation. Further, their influence on the permeability of co-administered common drugs (ketoprofen, paracetamol, metoprolol and verapamil) across Caco-2 monolayers was evaluated. The apical-to-basolateral permeability of highly permeable verapamil was mostly affected (decreased) during co-administration of the raspberry extract or the ellagitannin fraction. Ketoprofen permeability was decreased by the ellagitannin fraction. Consumption of food rich in phytochemicals, as demonstrated here with chemically characterized raspberry extract and fractions, with well-absorbing drugs would seem to affect the permeability of some of these drugs depending on the components. Thus their effects on the absorption of drugs in-vivo cannot be excluded. PMID- 17132219 TI - Enhanced delivery of nebulised salbutamol during non-invasive ventilation. AB - Non-invasive ventilation (NIV) is used to treat acute respiratory failure. Nebulised drugs can be delivered concurrently with NIV or during breaks from ventilatory support. We hypothesised that the amount of nebulised salbutamol inhaled when delivered via bi-level ventilation would be no different to the amount available directly from the same nebuliser. A standard bi-level ventilation circuit was attached to a lung model simulating adult respiration. Drug delivery was compared when salbutamol (5 mg) was nebulised at different positions in the circuit and separately, with no ventilator. The amount of salbutamol contained in various particle size fractions was also determined. Nebuliser position within the NIV circuit was critically important for drug delivery. Optimal delivery of salbutamol occurred with the expiration port between the facemask and nebuliser (647+/-67 micro g). This was significantly better than nebulisation without the ventilator (424+/-61 micro g; P < 0.01). Delivery when the nebuliser was positioned between the facemask and expiration port was 544+/-85 micro g. The amount of salbutamol contained in particles < 5 micro m was significantly increased when the nebuliser was used in conjunction with bi-level ventilation (576+/-60 micro g vs 300+/-43 micro g, P < 0.001). We conclude that nebulised bronchodilator therapy, using a Cirrus jet nebuliser, during bi-level ventilation increases respirable particles likely to be inhaled when the nebuliser is optimally positioned within the circuit. PMID- 17132220 TI - HIV protease inhibitors decrease VEGF/HIF-1alpha expression and angiogenesis in glioblastoma cells. AB - Glioblastomas are malignant brain tumors that are rarely curable, even with aggressive therapy (surgery, chemotherapy, and radiation). Glioblastomas frequently display loss of PTEN and/or epidermal growth factor receptor activation, both of which activate the PI3K pathway. This pathway can increase vascular endothelial growth factor (VEGF) and hypoxia-inducible factor (HIF) 1alpha expression. We examined the effects of two human immunodeficiency virus protease inhibitors, nelfinavir and amprenavir, which inhibit Akt signaling, on VEGF and HIF-1alpha expression and on angiogenesis. Nelfinavir decreased VEGF mRNA expression and VEGF secretion under normoxia. Downregulation of P-Akt decreased VEGF secretion in a manner similar to that of nelfinavir, but the combination of the two had no greater effect, consistent with the idea that nelfinavir decreases VEGF through the PI3K/Akt pathway. Nelfinavir also decreased the hypoxic induction of VEGF and the hypoxic induction of HIF-1alpha, which regulates VEGF promoter. The effect of nelfinavir on HIF-1alpha was most likely mediated by decreased protein translation. Nelfinavir's effect on VEGF expression had the functional consequence of decreasing angiogenesis in in vivo Matrigel plug assays. Similar effects on VEGF and HIF-1alpha expression were seen with a different protease inhibitor, amprenavir. Our results support further research into these protease inhibitors for use in future clinical trials for patients with glioblastoma multiformes. PMID- 17132221 TI - Apigenin suppresses cancer cell growth through ERbeta. AB - Two flavonoids, genistein and apigenin, have been implicated as chemopreventive agents against prostate and breast cancers. However, the mechanisms behind their respective cancer-protective effects may vary significantly. The goal of this study was to determine whether the antiproliferative action of these flavonoids on prostate (DU-145) and breast (MDA-MB-231) cancer cells expressing only estrogen receptor (ER) beta is mediated by this ER subtype. It was found that both genistein and apigenin, although not 17beta-estradiol, exhibited antiproliferative effects and proapoptotic activities through caspase-3 activation in these two cell lines. In yeast transcription assays, both flavonoids displayed high specificity toward ERbeta transactivation, particularly at lower concentrations. However, in mammalian assay, apigenin was found to be more ERbeta-selective than genistein, which has equal potency in inducing transactivation through ERalpha and ERbeta. Small interfering RNA-mediated downregulation of ERbeta abrogated the antiproliferative effect of apigenin in both cancer cells but did not reverse that of genistein. Our data unveil, for the first time, that the anticancer action of apigenin is mediated, in part, by ERbeta. The differential use of ERalpha and ERbeta signaling for transaction between genistein and apigenin demonstrates the complexity of phytoestrogen action in the context of their anticancer properties. PMID- 17132222 TI - Increases in c-Src expression level and activity do not promote the growth of human colorectal carcinoma cells in vitro and in vivo. AB - The levels and activity of c-Src in colorectal cancer cells increase steadily during the course of colorectal carcinogenesis and are most highly elevated in advanced metastatic disease. However, the effects of increases in c-Src activity on the proliferation of colorectal cancer cells during early and late stages of tumorigenesis remain elusive. To study the consequences of increases in c-Src levels and activity on the growth of colorectal cancer cells in later stages of colorectal carcinogenesis, we developed human colorectal cancer cell lines in which c-Src levels and activity could be inducibly increased by a tightly controlled expression of wild-type c-Src or of the constitutively active mutant of c-Src, c-SrcY527F. Src induction activated multiple signaling pathways (often associated with a proliferative response) but promoted neither cell proliferation in vitro nor tumor growth in a xenograft model in vivo. These results indicate that, in more advanced stages of colorectal carcinogenesis, increases in c-Src levels and activity are likely to have functions other than the direct promotion of tumor growth. PMID- 17132223 TI - Lentivirus-mediated silencing of Tiam1 gene influences multiple functions of a human colorectal cancer cell line. AB - T lymphoma invasion and metastasis 1 (Tiam1) is a metastasis-related gene of T lymphoma that is also involved in the metastasis of a variety of other cancers. In this study, we tested the hypothesis that Tiam1 is a determinant of proliferation and metastasis in colorectal cancer, and we examined the effect of the inhibition of Tiam1 expression on proliferation and metastasis. We succeeded in establishing the Tiam1 knockdown colorectal cancer cell line using human immunodeficiency virus lentivirus-mediated RNA interference (RNAi) and found that the silencing of Tiam1 resulted in the effective inhibition of in vitro cell growth and of the invasive ability of colorectal cancer cells. Using an orthotopic xenograft model in nude mice, we confirmed that Tiam1 silencing could reduce tumor growth by subcutaneous injection and could suppress lung and liver metastases of colorectal cancer cells. Our results suggest that Tiam1 truly plays a causal role in the metastasis of colorectal cancer and that RNAi-mediated silencing of Tiam1 may provide an opportunity to develop a new treatment strategy for colorectal cancer. PMID- 17132224 TI - Global expression-based classification of lymph node metastasis and extracapsular spread of oral tongue squamous cell carcinoma. AB - Regional lymph node metastasis is a critical event in oral tongue squamous cell carcinoma (OTSCC) progression. The identification of biomarkers associated with the metastatic process would provide critical prognostic information to facilitate clinical decision making for improved management of OTSCC patients. Global expressional profiles were obtained for 25 primary OTSCCs, where 11 cases showed lymph node metastasis (pN+) histologically and 14 cases were nonmetastatic (pN-). Seven of pN+ cases also exhibited extracapsular spread (ECS) of metastatic nodes. Multiple expression indices were used to generate signature gene sets for pN+/- and ECS+/- cases. Selected genes from signature gene sets were validated using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The classification powers of these genes were then evaluated using a logistic model, receiver operating characteristic curve analysis, and leave-one-out cross validation. qRT-PCR validation data showed that differences at RNA levels are either statistically significant (P < .05) or suggestive (P < .1) for six of eight genes tested (BMP2, CTTN, EEF1A1, GTSE1, MMP9, and EGFR) for pN+/- cases, and for five of eight genes tested (BMP2, CTTN, EEF1A1, MMP9, and EGFR) for ECS+/ cases. Logistic models with specific combinations of genes (CTTN+MMP9+EGFR for pN and CTTN+EEF1A1+MMP9 for ECS) achieved perfect specificity and sensitivity. Leave-one-out cross-validation showed overall accuracy rates of 85% for both pN and ECS prediction models. Our results demonstrated that the pN and the ECS of OTSCCs can be predicted by gene expression analyses of primary tumors. PMID- 17132225 TI - Immune escape for renal cell carcinoma: CD70 mediates apoptosis in lymphocytes. AB - Tumors can escape immune recognition and destruction through the induction of apoptosis in lymphocytes. Although renal cell carcinoma (RCC) is able to prevent immune recognition, only a few genes (such as FasL) that are relevant for RCC immune escape have been identified so far. We have previously shown that some apoptosis-inducing genes are overexpressed in RCC. We hypothesized that these genes could be part of the immune-escape strategy of these tumors. Here we report that CD70, a cytokine overexpressed in RCC, promotes lymphocyte apoptosis through interaction with its receptor CD27 and with the intracellular receptor-binding protein SIVA. Apoptosis increased after cocultivating lymphocytes with the RCC cell lines A498 and CAKI2. The addition of recombinant soluble CD70 to both native lymphocytes and a T-cell cell line resulted in increased lymphocyte apoptosis as well. Furthermore, induced apoptosis could be partially blocked with anti-CD27 and anti-CD70 antibodies. Our results strongly indicate a role for CD70 and CD27 receptor in lymphocyte apoptosis within the tumor environment. Apoptosis mediated by exposure to the CD70 secreted by tumor cells may contribute to the failure of RCC patients to develop an effective lymphocyte-mediated antitumor response. PMID- 17132226 TI - Inhibition of spontaneous breast cancer metastasis by anti-Thomsen-Friedenreich antigen monoclonal antibody JAA-F11. AB - Thomsen-Friedenreich antigen (TF-Ag) is expressed in many carcinomas, including those of the breast, colon, bladder, and prostate. TF-Ag is important in adhesion and metastasis and as a potential immunotherapy target. We hypothesized that passive transfer of JAA-F11, an anti-TF-Ag monoclonal antibody, may create a survival advantage for patients with TF-Ag-expressing tumors by cytotoxicity, blocking of tumor cell adhesion, and inhibition of metastasis. This was tested using in vitro models of tumor cell growth; cytotoxicity assays; in vitro, ex vivo, and in vivo models of cancer metastasis; and, finally, in vivo effects in mice with metastatic breast cancer. Unlike some anti-TF-Ag antibodies, JAA-F11 did not enhance breast carcinoma cell growth. JAA-F11 did not induce the killing of 4T1 tumor cells through complement-dependent cytotoxicity or apoptotic mechanisms. However, JAA-F11 blocked the stages of metastasis that involve the adhesion of human breast carcinoma cells to human endothelial cells (human umbilical vein endothelial cells and human bone marrow endothelial cells 60) in in vitro static adhesion models, in a perfused ex vivo model, and in murine lung vasculature in an in vivo metastatic deposit formation assay. JAA-F11 significantly extended the median survival time of animals bearing metastatic 4T1 breast tumors and caused a > 50% inhibition of lung metastasis. PMID- 17132227 TI - Inhibition of c-Met as a therapeutic strategy for esophageal adenocarcinoma. AB - The hepatocyte growth factor (HGF) receptor c-Met is a tyrosine kinase receptor with established oncogenic properties. We have previously shown that c-Met is usually overexpressed in esophageal adenocarcinoma (EA), yet the implications of c-Met inhibition in EA remain unknown. Three c-Met-overexpressing EA cell lines (Seg-1, Bic-1, and Flo-1) were used to examine the effects of a c-Met-specific small molecule inhibitor (PHA665752) on cell viability, apoptosis, motility, invasion, and downstream signaling pathways. PHA665752 demonstrated dose dependent inhibition of constitutive and/or HGF-induced phosphorylation of c-Met, which correlated with reduced cell viability and inhibition of extracellular regulated kinase 1/2 phosphorylation in all three EA cell lines. In contrast, PHA665752 induced apoptosis and reduced motility and invasion in only one EA cell line, Flo-1. Interestingly, Flo-1 was the only cell line in which phosphatidylinositol 3-kinase (PI3K)/Akt was induced following HGF stimulation. The PI3K inhibitor LY294002 produced effects equivalent to those of PHA665752 in these cells. We conclude that inhibition of c-Met may be a useful therapeutic strategy for EA. Factors other than receptor overexpression, such as c-Met dependent PI3K/Akt signaling, may be predictive of an individual tumor's response to c-Met inhibition. PMID- 17132228 TI - Role of the NEDD8 modification of Cul2 in the sequential activation of ECV complex. AB - ECV is an E3 ubiquitin ligase complex, which is composed of elongins B and C, Rbx1, Cul2, and the substrate-conferring von Hippel-Lindau (VHL) tumor-suppressor protein that targets the catalytic alpha subunit of hypoxia-inducible factor (HIF) for oxygen-dependent ubiquitin-mediated destruction. Mutations in VHL that compromise proper HIFalpha regulation through ECV have been documented in the majority of renal cell carcinomas, underscoring the significance of the VHL-HIF pathway in renal epithelial oncogenesis. Recent evidence has shown that the modification of Cul2 by the ubiquitin-like molecule NEDD8 increases the activity of ECV to ubiquitylate HIFalpha. However, the underlying mechanism responsible for the NEDD8-mediated induction of ECV function is unknown. Here, we demonstrate that oxygen-dependent recognition of HIFalpha by VHL triggers Rbx1-dependent neddylation of Cul2, which preferentially engages the E2 ubiquitin-conjugating enzyme UbcH5a. These events establish a central role for the neddylation of Cul2 in a previously unrecognized, temporally coordinated activation of ECV with the recruitment of its substrate HIFalpha. PMID- 17132229 TI - Signaling from p53 to NF-kappaB determines the chemotherapy responsiveness of neuroblastoma. AB - Neuroblastic (N) type neuroblastoma (NB) is the predominant cell type in NB tumors. Previously, we determined that activated nuclear factor kappaB (NF kappaB) is required for doxorubicin and etoposide to kill N-type NB cells. This study was undertaken to determine how NF-kappaB is activated by these agents. The results show that p53 protein levels increase within 15 to 30 minutes of treatment. This increase occurs before the degradation of inhibitor of NF-kappaB (I-kappaB) alpha and the NF-kappaB-dependent activation of gene transcription. Moreover, p53 is necessary for NF-kappaB activation because cells with inactive p53 were resistant to NF-kappaB-mediated cell death. This pathway was further defined to show that p53 leads to the activation of MAPK/ERK activity kinase (MEK) 1 through a process that depends on protein synthesis and H-Ras. MEK1, in turn, mediates I-kappaB kinase activation. Together, these results demonstrate for the first time how NF-kappaB is activated in NB cells in response to conventional drugs. Furthermore, these findings provide an explanation as to why H-Ras expression correlates with a favorable prognosis in NB and identify intermediary signaling molecules that are targets for discovering treatments for NB that is resistant to conventional agents. PMID- 17132242 TI - External contamination of hair with cocaine: evaluation of external cocaine contamination and development of performance-testing materials. AB - The National Laboratory Certification Program undertook an evaluation of the dynamics of external contamination of hair with cocaine (COC) while developing performance testing materials for Federal Drug-Free Workplace Programs. This characterization was necessary to develop performance materials that could evaluate the efficacy of hair testing industry's decontamination procedures. Hair locks (blonde to dark brown/black) from five different individuals were contaminated with cocaine HCl. Hair locks were then treated with a synthetic sweat solution and hygienic treatments to model real-life conditions. Hair locks were shampooed daily (Monday through Friday) for 10 weeks, and samples of the hair locks were analyzed for COC, benzoylecgonine (BE), cocaethylene (CE), and norcocaine (NCOC). Three commercial analytical laboratories analyzed samples under three protocols: no decontamination procedure, individual laboratory decontamination, or decontamination by an extended buffer procedure at RTI International. Results indicated substantial and persistent association of all four compounds with all hair types. Hair that was not decontaminated had significantly greater quantities of COC and BE than did hair that was decontaminated. The only hair samples below detection limits for all four compounds were those decontaminated 1 h after contamination. Additionally, BE/COC ratios increased significantly over the 10-week study (regardless of decontamination treatment). From 21 days postcontamination until the end of the study, the mean BE/COC ratio for all hair types exceeded 0.05, the proposed Federal Mandatory Guidelines requirement. The largest variability in results was observed for samples decontaminated by participant laboratories. This suggests that current laboratory decontamination strategies will increase variability of performance testing sample results. None of the decontamination strategies used in the study were effective at removing all contamination, and some of the contaminated hair in this study would have been reported as positive for cocaine use based on the proposed Federal Mandatory Guidelines. PMID- 17132243 TI - Major and minor metabolites of cocaine in human plasma following controlled subcutaneous cocaine administration. AB - Cocaine is rapidly metabolized to major metabolites, benzoylecgonine (BE) and ecgonine methyl ester (EME), and minor metabolites, norcocaine, p-hydroxycocaine, m-hydroxycocaine, p-hydroxybenzoylecgonine (pOHBE), and m-hydroxybenzoylecgonine. This IRB-approved study examined cocaine and metabolite plasma concentrations in 18 healthy humans who provided written informed consent to receive low (75 mg/70 kg) and high (150 mg/70 kg) subcutaneous cocaine hydrochloride doses. Plasma specimens, collected prior to and up to 48 h after dosing, were analyzed by gas chromatography-mass spectrometry (2.5 ng/mL limits of quantification). Cocaine was detected within 5 min, with mean+/-SE peak concentrations of 300.4+/-24.6 ng/mL (low) and 639.1+/-56.8 ng/mL (high) 30-40 min after dosing. BE and EME generally were first detected in plasma 5-15 min post-dose; 2-4 h after dosing, BE and EME reached mean maximum concentrations of 321.3+/-18.4 (low) and 614.7+/ 46.0 ng/mL (high) and 47.4+/-3.0 (low) and 124.4+/-18.2 ng/mL (high), respectively. Times of last detection were BE>EME>cocaine. Minor metabolites were detected much less frequently for up to 32 h, with peak concentrations 90% of morphine and hydromorphone from their glucuronide standards but enzyme hydrolysis had lower and variable efficiency, depending on the opiate type and the enzyme source. In patient specimens, much higher concentrations of free codeine, morphine, hydromorphone, and oxymorphone were obtained with acid hydrolysis than with various enzyme methods. Incomplete hydrolysis using beta glucuronidase could lead to false-negative results for many opioids when urine is tested for drugs of abuse. We conclude that acid hydrolysis is the method of choice for GC-MS confirmation of urine opioids. PMID- 17132255 TI - A first look at duloxetine (Cymbalta) in a postmortem laboratory. AB - Duloxetine (Cymbalta) is manufactured by Eli Lilly and Company and is the newest antidepressant to be approved by the Food and Drug Administration (FDA). Duloxetine is a potent serotonin and norepinephrine reuptake inhibitor that is also used for the management of pain associated with diabetic peripheral neuropathy. With the introduction of any new drug, toxicology laboratories around the nation experience the same problems: lack of information about the chemical and physical properties of the new drug, detection methodologies from biological specimens, and interpretation of quantitative values. Since its FDA approval in 2002, the Los Angeles County Department of Coroner Toxicology Laboratory has detected and quantitated duloxetine in 12 postmortem cases. The isolation of duloxetine from postmortem specimens consisted of a basic, liquid-liquid (n butylchloride) extraction procedure. Duloxetine was detected in our general, pharmaceutical, basic drugs screen that utilizes gas chromatography-nitrogen phosphorus detection (GC-NPD) and GC-mass spectrometry (MS), and the quantitation was specifically by GC-MS. Linearity was achieved from 0.05 to 3.0 mg/L with the limit of detection at 0.03 mg/L. Presented are the case histories, demographics, cause/manner of death, and the postmortem tissue distribution ranges of duloxetine: central blood, not detected (ND)-0.59 mg/L (12 cases); femoral blood, ND-0.26 mg/L (9 cases); vitreous humor, ND-0.23 mg/L (4 cases); liver, 0.28-22 mg/kg (8 cases); gastric contents, 0.08-86 mg total (6 cases); bile, 0.57-3.1 mg/L (7 cases); and urine, 0.07-0.47 mg/L (6 cases). The detection and quantitation of duloxetine in these 12 case studies are considered the first to be reported in the literature; all are designed to aid the forensic toxicologist with the interpretation of his/her own casework. PMID- 17132256 TI - Performance evaluation of thermal desorption system (TDS) for detection of basic drugs in forensic samples by GC-MS. AB - Stir bar sorptive extraction is an innovative sample extraction technique that can be used to process blood, urine, and tissue samples for routine drug screening in the forensic toxicology laboratory. The Gerstel Twister desorption unit (TDU) system is a multifunctional desorption unit capable of determining the presence of analytes from liquid samples after extraction using the Twister stir bar. The TDU desorption system was evaluated for use in combination with gas chromatography-mass spectrometry (GC-MS) for determining the presence of basic drugs in forensic samples. Human blood fortified with known quantities of drugs was used to evaluate sample diluents, extraction time, injection parameters and recovery. Case specimens containing drugs typically encountered in forensic samples were evaluated using the desorption method and compared with a liquid liquid extraction method followed by GC-MS analysis. This evaluation demonstrated that the TDU desorptive method worked equally as well as the routine extraction method for the detection of basic drugs in screening forensic samples. In addition, the described technique avoids the use of extraction solvents and the subsequent centrifugation, transfer, and concentration steps required of liquid liquid and solid-phase extraction methods. PMID- 17132257 TI - Liquid chromatography-tandem mass spectrometry for the determination of colchicine in postmortem body fluids. Case report of two fatalities and review of the literature. AB - Poisoning by colchicine may occur following ingestion of this alkaloid used for the treatment of acute gouty arthritis. The authors report two fatalities and describe a liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS) triple-quadrupole method for the determination of colchicine in autopsy samples. One milliliter of heart blood, femoral blood, urine, bile, gastric, and vitreous each were extracted with saturated NH4Cl at pH 9.6 and dichloromethane/5% isopropanol. Separation was achieved on a C18-Xterra column with a mobile phase consisting of 2 mM ammonium formate buffer (pH 3)/acetonitrile in a gradient mode. Four product ions of the protonated molecule were monitored. The method was fully validated in whole blood (1 mL) and was linear in the range of 0.5-50 ng/mL (r2>0.99). The limit of detection was 0.1 ng/mL (50 times S/N), and the limit of quantitation was 0.5 ng/mL with RSDs<11.8% intraday (n=6), <18.7% interday (n=18), and accuracy<3% (n=18). Case #1: a 33 year-old nurse committed suicide by the ingestion of 80 colchicine 1-mg tablets. She died 61 h later after resuscitation procedures. Colchicine was found in heart blood at 5.2 ng/mL, femoral blood at 17.4 ng/mL, urine at 19.4 ng/mL, bile at 42.8 ng/mL, gastric at 348 ng/mL, and vitreous at 3 ng/mL. Case #2: a 57-year-old man with gout was found dead at home. Colchicine was found in heart blood at 22.8 ng/mL, femoral blood at 21.9 ng/mL, lung blood at 45.2 ng/mL, urine at 148.5 ng/mL, bile at 1818.5 ng/mL, gastric at 219.8 ng/mL, and vitreous at 0.5 ng/mL. These results were consistent with death. Because of its good sensitivity, this LC-ESI-MS-MS triple-quadrupole method is suitable for the determination of colchicine not only in fatalities but also for pharmacokinetic studies. PMID- 17132258 TI - Topiramate-positive death-investigation and impaired-driving cases in Washington State. AB - Topiramate (Topamax), an effective seizure disorder treatment, received additional FDA approval for prevention of migraine headaches in August 2004 and has gained attention for its off-label uses, including psychiatric and eating disorders, neuropathic pain, and alcohol and drug dependency. Side effects of sedation, dizziness, ataxia, speech difficulty, nystagmus, paresthesia, and metabolic acidosis are described. The manufacturer reports that tolerance to the antiseizure properties does not develop. With its established efficacy for epilepsy treatment and its increased use for other disorders, topiramate-positive findings are more common in death-investigation and human-performance casework. To evaluate the role of topiramate, we reviewed all topiramate-positive cases from our laboratory between 1998 and 2004, which constituted 132 cases (63 death investigations, 68 suspected impaired drivers, and 1 sexual assault case). The subjects were predominantly female (69%) with a mean and median age of 42. Blood topiramate concentrations ranged from 1 to 180 mg/L (median 6.4 mg/L, mean 8.4 mg/L), and 94% were positive for at least one additional drug. There was evidence of psychomotor impairment in some drivers with blood concentrations within the normal therapeutic range, and deaths attributed to topiramate alone occurred at concentrations as low as 50 mg/L. PMID- 17132259 TI - Fentanyl-related deaths in Ontario, Canada: toxicological findings and circumstances of death in 112 cases (2002-2004). AB - In order to characterize fentanyl-related deaths in the province of Ontario, Canada, a retrospective study of all cases in which fentanyl was quantitated in blood was conducted for the time period between 2002 and 2004. A total of 112 fentanyl-related deaths were identified. Decedents ranged in age from 4 to 93 years and comprised 63 men and 49 women. A variety of routes of administration of the drug were identified: transdermal application of Duragesic patches, intravenous injection of patch contents or fentanyl citrate solution, oral/transmucosal administration, and volatilization and inhalation of Duragesic systems. Blood fentanyl concentrations were determined for all modes of drug administration and are provided. There were 54 cases in which death was attributed solely to fentanyl intoxication; the mean blood concentration was 25 microg/L (range: 3.0-383 microg/L). This concentration range overlapped with blood fentanyl concentrations measured among cases where the presence of the drug was considered incidental. For example, a mean blood concentration of 12 microg/L was observed among 12 cases of natural death (range: 2.7-33 microg/L). Detailed case reports of six individuals are also included and provide additional insight into the use of this drug for both therapeutic and illicit means. PMID- 17132260 TI - Caffeine content of decaffeinated coffee. AB - Caffeine is the most widely consumed drug in the world with coffee representing a major source of intake. Despite widespread availability, various medical conditions necessitate caffeine-restricted diets. Patients on certain prescription medications are advised to discontinue caffeine intake. Such admonition has implications for certain psychiatric patients because of pharmacokinetic interactions between caffeine and certain anti-anxiety drugs. In an effort to abstain from caffeine, patients may substitute decaffeinated for caffeinated coffee. However, decaffeinated beverages are known to contain caffeine in varying amounts. The present study determined the caffeine content in a variety of decaffeinated coffee drinks. In phase 1 of the study, 10 decaffeinated samples were collected from different coffee establishments. In phase 2 of the study, Starbucks espresso decaffeinated (N=6) and Starbucks brewed decaffeinated coffee (N=6) samples were collected from the same outlet to evaluate variability of caffeine content of the same drink. The 10 decaffeinated coffee samples from different outlets contained caffeine in the range of 0-13.9 mg/16-oz serving. The caffeine content for the Starbucks espresso and the Starbucks brewed samples collected from the same outlet were 3.0-15.8 mg/shot and 12.0-13.4 mg/16-oz serving, respectively. Patients vulnerable to caffeine effects should be advised that caffeine may be present in coffees purported to be decaffeinated. Further research is warranted on the potential deleterious effects of consumption of "decaffeinated" coffee that contains caffeine on caffeine restricted patients. Additionally, further exploration is merited for the possible physical dependence potential of low doses of caffeine such as those concentrations found in decaffeinated coffee. PMID- 17132261 TI - Recovery of drugs of abuse from the Immunalysis Quantisal oral fluid collection device. AB - Drug recovery from a new oral fluid collection device was assessed. The evaluation was performed in vitro at three physiologically relevant concentrations for the following substances: amphetamine, methamphetamine, morphine, codeine, cocaine, benzoylecgonine, methadone, oxazepam, and Delta(9) tetrahydrocannabinol (THC). Drug-free and drug-fortified controls were prepared and their concentration verified by liquid chromatography-tandem mass spectrometry. Aliquots of the controls were then "collected" with the device (n=3) using the manufacturer's recommended procedure. Collected samples were stored for 12 h to simulate shipping before analysis. Fresh, non-"collected" aliquots of each pool (n=3) were concurrently analyzed. The drug recoveries from the Quantisal were expressed as a mean percentage of the concurrently analyzed aliquots that were not subjected to device collection. Recoveries for oxazepam exceeded 97%, for amphetamine and methamphetamine exceeded 93%, and for opioids all exceeded 91%. The recoveries of cocaine were >91% and >82% for its polar metabolite, benzoylecgonine. Especially noteworthy was the recovery of THC from the Quantisal collector (81.3-91.4%). When compared with recoveries reported from other collection devices, the Quantisal was clearly superior. PMID- 17132262 TI - Validation of an ELISA method for screening methadone in postmortem blood. AB - In this study, we evaluate Venture Labs' enzyme-linked immunosorbent assay (ELISA) for the detection of methadone in postmortem specimens. Sixty-one postmortem specimens that previously screened positive for methadone along with 59 specimens which screened negative for methadone were included. All specimens were screened using the Venture Labs methadone assay in conjunction with a liquid liquid basic extraction and gas chromatographic-mass spectrometric (GC-MS) analysis. All cases screening positive by either method were confirmed for methadone and its metabolite 2-ethylidene-1,5-dimethyl- 3,3-diphenylpyrrolidine by a solid-phase extraction utilizing deuterated internal standards and GC-MS SIM. Twenty-four postmortem samples that screened negative by both methods were also extracted and analyzed using the confirmation method to demonstrate the validity of both screening methods. The intra- and interassay precision for the ELISA method was evaluated at the cut-off concentration used for the analysis (50 ng/mL). True positives, true negatives, false positives, and false negatives were calculated for the ELISA results as compared to the GC-MS screening data. The Venture Labs methadone assay demonstrated a sensitivity of 96.7%+/-2.3% and a specificity of 98.3%+/-1.7% relative to the GC-MS method. PMID- 17132263 TI - Multi-element screening by ICP-MS of two specimens of Napoleon's hair. AB - Since 1960, it has been demonstrated by various analytical procedures that high concentrations of arsenic were present in Napoleon's hair. Various authors, indicating that the detected arsenic levels are a consequence of external contamination, have challenged the results of these examinations. In order to shed more light on this historical controversy, we have tested two samples of Napoleon's hair by inductively coupled plasma-mass spectrometry (ICP-MS). The samples of hair were decontaminated with acetone and were cut into small segments. For multi-element screening, hair samples were mineralized in concentrated nitric acid for 1 h at 70 degrees C, diluted 1:40 in specific solution with rhodium as an internal standard, and finally analyzed by ICP-MS on a Thermo Electron ICP/MS X7. Multi-element analysis of Napoleon's hair samples revealed massive amounts of arsenic (42.1 and 37.4 ng/mg), antimony (2.1 and 1.8 ng/mg) and elevated levels of mercury (3.3 and 4.7 ng/mg) and lead (229 and 112 ng/mg). In the case of arsenic, these concentrations, 40 times higher than the normal values, confirm the hypothesis of a significant exposure to arsenic. The concentrations of the other elements, in particular antimony and mercury, are in agreement with the data already known about the therapeutic treatments given to Napoleon, which were based on calomel (salt of mercury) and tartar emetic (antimony). PMID- 17132264 TI - Investigation of a fatality due to diesel fuel No. 2 ingestion. AB - This paper presents a simple, rapid, reliable, and validated analytical method suited for forensic examination of diesel fuel No. 2 in biological specimens. The proposed methodology has been applied to the investigation of a forensic case with diesel fuel No. 2 ingestion. Case history and pathological and toxicological findings are described here to illustrate the toxicity of this complex hydrocarbon mixture. The toxicological significance and the possible mechanisms leading to death are also discussed. The toxicological initial screening and quantitation were performed by means of gas chromatography with flame-ionization detection and confirmation was performed using gas chromatography-mass spectrometry in total ion chromatogram mode. n-Tetradecane peak was selected to estimate diesel fuel No. 2 in all biological samples. Diesel fuel No. 2 analytical methodology was validated at five concentration levels from 5 to 400 mg/L. The method provided extraction recoveries between 89.0% and 97.9%. The limit of detection was 1 mg/L and the limit of quantitation was 5 mg/L. The linearity of the blood calibration curves was excellent with r2 values of >0.999. Intraday and interday precisions had a coefficient of variation 50 microM), as ascorbate has been shown to regenerate tocopherol from its oxidised tocopheroxyl radical form in vitro. Healthy male subjects received alpha-tocopherol supplements (400 IU RRR-alpha-tocopherol/day for 6 weeks) in a placebo-controlled, double-blind intervention study. There were no significant differences in monocyte CD11b expression, monocyte adhesion to endothelial cells, plasma C-reactive protein or sICAM-1 concentrations post-supplementation. There was no evidence for nuclear translocation of NF-kappaB in isolated resting monocytes, nor any effect of alpha-tocopherol supplementation. However, post supplementation, sVCAM-1 levels were decreased in all subjects and sE-selectin levels were increased in the vitamin C-replete group only; a weak positive correlation was observed between sE-selectin and alpha-tocopherol concentration. In conclusion, alpha-tocopherol supplementation had little effect on cardiovascular disease risk factors in healthy subjects and the effects of tocopherol were not consistently affected by plasma vitamin C concentration. PMID- 17132271 TI - Differential regulation of intracellular redox state by extracellular matrix proteins in glomerular mesangial cells: potential role in diabetic nephropathy. AB - Advanced diabetic nephropathy is characterized by abnormal synthesis of extracellular matrix (ECM) proteins, such as collagen I (COL I). The present experiments were designed to test the hypothesis that the presence of abnormal ECM proteins may be responsible for increased generation of reactive oxygen species (ROS) that are thought to have an important role in the pathogenesis of diabetic nephropathy. SV40 MES 13 murine mesangial cells were plated on COL I or collagen IV (COL IV) for 3 h at 5.5 or 25 mM D-glucose concentration. Increased intracellular ROS generation and reduced intracellular nitric oxide (NO) production was measured in cells attached to COL I compared with cells attached to COL IV. Treatment with N(omega)-nitro-L-arginine methyl ester hydrochloride (L NAME), an inhibitor of NO synthase, reduced this difference in ROS generation between cells attached to either COL I or IV. The results using antibodies against integrins also indicated that an alpha(2) integrin-mediated pathway was involved in the different response in ROS generation caused by ECM proteins. These results suggest that contact between altered ECM proteins that are present in advanced diabetic nephropathy and mesangial cells has the potential to increase intracellular oxidative stress, leading to progressive glomerular damage. PMID- 17132272 TI - Yeast flavohemoglobin protects against nitrosative stress and controls ferric reductase activity. AB - The role of Saccharomyces cerevisiae flavohemoglobin (Yhb1) is controversial and far from understood. This study compares the effects of nitrosative and oxidative challenge on the yeast mutant lacking the YHB1 gene. Growth of the mutant was impaired by nitrosoglutathione and peroxynitrite, whereas increased sensitivity to reactive oxygen species was not observed. Increased levels of intracellular NO(*) after incubation with NO(*) donors were found in the mutants cells as compared to the wild-type cells. Deletion of the YHB1 gene was found to augment the reduction of Fe(3+) by yeast cells which suggests that flavohemoglobin participates in regulation of the activity of plasma membrane ferric reductase(s). PMID- 17132273 TI - The Human Tissue Act: implications for clinical biochemistry. AB - The Human Tissue Act sets out a new legal framework for the use and retention of tissues from living persons as well as the removal, retention and use of tissue and organs from the deceased. The Human Tissue Authority (HTA) has been established as the regulatory body in relation to the Act and also to give advice and guidance initially through the publication of Codes of Practice. The HTA will also license activities within the remit of the Act and will inspect to ensure compliance with the Act and licence conditions. The Act establishes consent as the essential requirement for the lawful retention and use of tissues and organs; failure to comply could result in penalties that include a custodial sentence. In addition to solid organs and tissues, the Act applies to the use of other specimens including body fluids if they contain cells. Tissue can be stored and used without consent for a number of purposes on the basis that these are integral to the general provision of clinical and diagnostic services. These include clinical audit, education or training relating to human health, performance assessment and quality assurance. In the case of research, the Act allows tissue to be used without consent, provided that the tissue is anonymized so that the researcher cannot identify from whom the material came. Linking with medical records is allowed, provided patient-identifying information is not obtained. There is a requirement to respect the wishes of any patient who specifies that they do not wish diagnostic or therapeutic samples to be kept or used for additional purposes. PMID- 17132274 TI - Thyroid hormone resistance. AB - Resistance to thyroid hormone (RTH) is a rare autosomal dominant inherited syndrome of reduced end-organ responsiveness to thyroid hormone. Patients with RTH have elevated serum free thyroxine (FT4) and free triiodothyronine (FT3) concentrations and normal or slightly elevated serum thyroid stimulating hormone (TSH) level. Despite a variable clinical presentation, the common characteristic clinical features are goitre but an absence of the usual symptoms and metabolic consequences of thyroid hormone excess. Patients with RTH can be classified on clinical grounds alone into either generalized resistance (GRTH), pituitary resistance (PRTH) or combined. Mutations in the thyroid hormone receptor (TR) beta gene are responsible for RTH and 122 different mutations have now been identified belonging to 300 families. With the exception of one family found to have complete deletion of the TRbeta gene, all others have been demonstrated to have minor alterations at the DNA level. The differential diagnosis includes a TSH-secreting pituitary adenoma and the presence of endogenous antibodies directed against thyroxine (T4) and triiodothyronine (T3). Failure to differentiate RTH from primary thyrotoxicosis has resulted in the inappropriate treatment of nearly one-third of patients. Although occasionally desirable, no specific treatment is available for RTH; however, the diagnosis allows appropriate genetic counselling. PMID- 17132275 TI - Diagnosis and screening for familial hypercholesterolaemia: finding the patients, finding the genes. AB - Familial hypercholesterolaemia (FH) is a genetic disorder in which the concentration of serum cholesterol is elevated from birth and leads to premature coronary heart disease. FH is commonly caused by a mutation in the LDL receptor, but mutations in other genes can lead to a phenotype similar to FH. FH exhibits marked phenotypic variability due to genetic, metabolic and environmental factors. The presence of tendon xanthomata is the characteristic clinical sign seen in many patients with FH, but they may also have other non-specific signs of lipid disorders such as corneal arcus and xanthelasmata. Premature vascular disease is apparent in many patients. The wide variety of mutations and phenotypic variability have made it difficult to establish definite diagnostic criteria, but three sets of clinical criteria commonly used are the Simon Broome criteria, the Dutch Lipid Clinic criteria and the American criteria. FH screening fits the Wilson and Jungner recommendations for validity of a screening programme. Screening could be carried out on a population basis, in a clinical setting or by application to relatives of probands. This latter approach, termed cascade testing, appears to be the more cost-effective compared with population screening and can be carried out using clinical criteria or genetic testing, or by a combination of both methods. Clinicians need to be made more aware of the clinical features of FH and how to diagnose it in order to increase the index of suspicion and instigate appropriate treatment early, with the aim of preventing premature coronary heart disease. PMID- 17132276 TI - Current perspectives in protein array technology. AB - This article reviews post-2000 trends in the development of two-dimensional protein microarrays and nanoarrays. Progress in array manufacture, assay design and applications are considered, with an emphasis on issues surrounding the implementation of arrays in clinical diagnostics. These include the effect of factors in the pre-analytical phase (quality of the reagents, sample integrity, etc.), and those in the analytical phase that contribute to inaccuracy and imprecision of an array-based assay. Important requirements for the quality control and quality assurance of protein microarray assays as they move from the research environment into routine clinical application are also discussed. PMID- 17132277 TI - Vitamin D concentrations in an UK inner-city multicultural outpatient population. AB - BACKGROUND: Vitamin D deficiency is still thought to be widespread in the UK and in recent years the number of cases of rickets reported in children has increased. In this study, the distribution of vitamin D and the prevalence of vitamin D deficiency have been determined for a multi-ethnic population from the inner-city area of Birmingham, UK, where a vitamin D testing service has been readily available for over 10 years. METHODS: Serum 25-hydroxyvitamin D concentration was determined using an automated platform (Nichol's Advantage Speciality System) for 830 outpatient samples collected randomly at the end of summer (September). RESULTS: In our total study population, prevalence of vitamin D deficiency, defined as a 25-hydroxyvitamin D concentration < 10 microg/L, was high (24%): one in eight Caucasians, one in four Black Afro-Caribbeans and one in three Asians were found to be deficient. Levels of deficiency were much higher in Asian women, with almost one in two individuals (43%) found to have a vitamin D level below 10 microg/L. CONCLUSION: Our study has shown that widespread vitamin D deficiency in a UK inner-city population remains an issue. In concordance with other studies, we found a high prevalence of vitamin D deficiency in Afro Caribbean and Asians, and, in particular, women. It is clear that more routine screening of vitamin D is needed. PMID- 17132278 TI - Simultaneous quantitation of homocysteine, cysteine and methionine in plasma and urine by liquid chromatography-tandem mass spectrometry. AB - BACKGROUND: A method utilizing liquid chromatography-electrospray tandem mass spectrometry (LC-MS/MS) has been developed and evaluated for the determination of total homocysteine, cysteine and methionine in plasma and urine. The simultaneous measurement of homocysteine and methionine concentrations may help explain the underlying mechanism responsible for hyperhomo-cysteinaemia. METHODS: Samples were prepared by simple protein precipitation after reduction of disulphides by dithiothreitol. Reduced analyte signal caused by ionization suppression effects, seen with plasma samples, was compensated for with matrix-matched standards, and the use of isotopically labelled internal standards. Recovery for each analyte was better than 94%. RESULTS: Concentrations of plasma homocysteine determined by LC-MS/MS were compared with those obtained by two automated commercially available FDA-approved procedures: (i) high-performance liquid chromatography (HPLC) with pre-column derivatization and fluorescence detection and (ii) by fluorescence polarization immunoassay (FPIA). Agreement with the LC-MS/MS method is given by the Deming regression equations LC-MS/MS = 1.062 HPLC-0.01 and LC MS/MS = 1.104 FPIA-0.43. CONCLUSION: Low reagent costs together with the relative simplicity of sample preparation make the LC-MS/MS method well suited, not only for research work but also in those laboratories with a tandem mass spectrometer, for the measurement of routine clinical samples. PMID- 17132279 TI - Comparison of Bayer Advia Centaur immunoassay results obtained on samples collected in four different Becton Dickinson Vacutainer tubes. AB - BACKGROUND: Medicines and Healthcare products Regulatory Agency's (MHRA's) Medical Device Alert MDA/2004/048 described bias in some endocrine test results obtained on a few immunoassay platforms, particularly the Bayer Advia Centaur instrument, when using blood specimens collected into Becton Dickinson (BD) Vacutainer SSTII Advance tubes. As users of BD tubes and the Advia Centaur instrument, we addressed our concerns about the quality of the results that we had previously reported by undertaking an independent study. METHOD: We compared the results of 15 immunoassays performed on Bayer Advia Centaur using blood specimens collected into four different BD Vacutainer tubes (plain, old and newly released BD SSTII Advance, and BD PSTII). RESULTS: Compared with plain tubes, old SSTII Advance tube results showed no bias for testosterone, CA15-3, follicle stimulating hormone and folate assays, but gave a positive bias for cortisol and a negative bias for vitamin-B12. Compared with plain tubes, BD PSTII tubes gave no significant bias for thyroid function tests, prolactin, parathyroid hormone, and CA125, but gave a negative bias for steroid assays, and a positive bias for gonadotrophins. The results obtained using new BD SSTII Advance tubes were generally comparable with those on plain tubes. CONCLUSIONS: Only for cortisol did our findings support the bias described by MHRA. Based on our results, apart from vitamin-B12 and possibly cortisol, there may have been no significant influence on clinical decisions as a result of using the old BD SSTII Advance specimen tubes. New BD SSTII Advance tubes and plain tubes give generally comparable results. BD PSTII tubes should not be used for steroid hormone measurements on the Bayer Advia Centaur instrument. PMID- 17132280 TI - Sodium measurement: effects of differing sampling and analytical methods. AB - BACKGROUND: It is recommended that children receiving intravenous fluids should have frequent biochemical monitoring, in some situations 4-6 hourly. Small changes in sodium must be detected, requiring very high precision from sodium analyses. Some children are monitored using venous blood analysed by indirect ion selective electrode (ISE) interchangeably with capillary blood analysed by direct ISE. Our aim was to determine whether variability in sample collection together with variability in sodium measurement would lead to results which were unacceptable in the clinical setting. METHODS: Fifty-seven adults had capillary and venous blood analysed for sodium using direct ISE and venous plasma analysed for sodium using indirect ISE. RESULTS: Comparison of capillary blood analysed by direct ISE with venous plasma analysed by indirect ISE demonstrated wide scatter and poor correlation of results: r = 0.36, standard deviation (SD) of the differences 2.7 mmol/L and range of limits of agreement 10.6 mmol/L. Significant biases were observed comparing capillary blood sodium with venous plasma sodium (P < 0.001), and comparing direct ISE with indirect ISE (P < 0.001). CONCLUSIONS: Venous plasma using indirect ISE and capillary blood with direct ISE cannot be used interchangeably to detect small changes in plasma sodium concentrations. To avoid misinterpretation of results when monitoring sodium over short time periods, the use of single methods of sampling and analysis must be strongly encouraged. PMID- 17132281 TI - Evaluation of the albumin cobalt binding (ACB) assay for measurement of ischaemia modified albumin (IMA) on the Beckman Coulter LX-20. AB - BACKGROUND: In the presence of ischaemia, albumin undergoes changes resulting in the formation of ischaemia-modified albumin (IMA). Increased serum concentrations of IMA have been found in patients with myocardial ischaemia. The purpose of this study was threefold: to evaluate the albumin cobalt binding (ACB) assay for measurement of IMA on the Beckman Coulter LX-20; to establish a reference range for IMA; and to investigate the relationship between IMA and total albumin concentrations. METHODS: The ACB assay was evaluated under the following headings: imprecision, accuracy and reliability. A reference range was established on a population of 81 healthy subjects. RESULTS: The within-batch coefficient of variation (CV) at IMA concentrations of 88, 99 and 120 KU/L were 1.4, 2.0 and 2.5%, respectively. The between-batch CVs at 74, 84 and 123 KU/L were 3.4, 3.3 and 3.0%, respectively. Comparison with the Cobas Mira Plus showed a mean negative bias of 7 KU/L. The 97.5th percentile established on our reference population was 110 KU/L. A significant inverse relationship was found between total serum albumin and IMA concentrations (r = -0.66, P < 0.0001). Correcting the IMA concentrations for total albumin in our reference population, using a formula devised in this study, yielded a range similar to that of uncorrected IMA. CONCLUSIONS: The ACB assay was found to have acceptable precision and performed very satisfactorily on the Beckman Coulter LX-20. A correction to measured IMA concentrations, to take into account total albumin concentrations, may need to be applied for the proper interpretation of IMA results. PMID- 17132282 TI - Effects of storage at -20 degrees C on ischaemia-modified albumin results. AB - BACKGROUND: Ischaemia-modified albumin (IMA) is being studied as a new marker for reversible ischaemia in patients presenting with possible cardiac chest pain. The conditions under which samples are stored prior to analysis may be critical in influencing the analytical result and hence the cut-off used in any particular study. METHODS: Sixty-eight samples taken during a study assessing the performance of IMA for risk stratification in patients presenting with possible cardiac chest pain were analysed both within 2.5 h of collection and after periods of storage at -20 degrees C. RESULTS: Samples stored at -20 degrees C yielded IMA values on average 3 units higher than those analysed within 2.5 h (mean 90.5 vs. 87.5; P < 0.00001). A Bland-Altman plot showed that the difference was not concentration dependent. CONCLUSIONS: These results indicate that decision cut-offs will be influenced by conditions of sample storage prior to IMA analysis, and that these should be stated in detail for each study. PMID- 17132283 TI - Serum calcium, urine calcium and polymorphisms of the calcium sensing receptor gene. AB - BACKGROUND: Recent studies have suggested a correlation between the A986S polymorphism of the calcium sensing receptor (CASR), and serum total and ionized calcium. This study aimed to assess the prevalence of three CASR polymorphisms in a West of Scotland population and relate genotype to serum and urine calcium levels. METHODS: Fasting blood and urine samples were obtained from 121 healthy male and female volunteers aged 20-60 years. Volunteers were genotyped for the A986S, Q1011E and R990G polymorphisms using allele-specific amplification and amplification-created restriction site techniques. Total calcium, ionized calcium and urine calcium excretion were measured using automated clinical chemistry analysers. RESULTS: Genotype frequencies for the A986S polymorphism were: AA, 74.4%; AS, 24.8%; SS, 0.8%. There was a small but statistically significant (P < 0.01) increase in ionized calcium concentration in AS individuals compared with the wild type (1.22 versus 1.20 mmol/L). No statistical difference was detected in serum total calcium or parameters of urine calcium excretion. Genotype frequencies for the remaining polymorphisms were: RR, 82.6%; RG, 16.5%; GG, 0.8% and QQ, 93.4%; QE, 6.6%; EE, 0%. Biochemical parameters in these individuals were not statistically different from the wild type. CONCLUSION: The increase in serum ionized calcium in the AS group was small and, therefore, unlikely to be of clinical significance. PMID- 17132284 TI - Effect of variability in assay bias and imprecision on external quality assessment bias and imprecision measures. AB - BACKGROUND: External quality assessment/proficiency testing programmes that report consensus means and coefficients of variations (CVs) are a potential source of information on assay bias and imprecision. This study examined the effect of variability in assay bias and imprecision on consensus means and CVs, using a computerized spreadsheet model. METHODS: A model with varying assay bias (mean and standard deviation) and assay imprecision (mean and standard deviation) was developed using a MS Excel 2003 spreadsheet with a macro to generate pseudo random numbers from a Gaussian distribution. The means, standard deviations and CVs of these data points were considered to simulate the consensus measures reported in external quality assessment testing programmes. RESULTS: The simulated consensus CV was very sensitive to increases in assay bias variability, with greater effects at low levels of mean assay imprecision. Changes in assay imprecision variability had a much smaller effect on the simulated consensus CV, while the simulated consensus mean was relatively insensitive to any change in the other variables. CONCLUSION: Consensus imprecision measures from external quality assessment programmes are unreliable measures of assay imprecision. Consensus means are relatively unaffected by variability in assay bias and imprecision, and can be used as reliable measures of assay bias from a statistical standpoint. PMID- 17132285 TI - Cellular content in plasma of Becton-Dickinson lithium-heparin tubes: cause of unreliable results in the IFCC-recommended lactate dehydrogenase method. AB - BACKGROUND: Measurement of lactate dehydrogenase (LDH) with the International Federation of Clinical Chemistry (IFCC)-recommended method in plasma was reported unreliable with primary evacuated lithium-heparin tubes from Becton-Dickinson, despite adequate removal of platelets from the plasma by optimized sample preparation. In this study, the composition of the top layer of the heparin plasma was investigated in relation to these duplicate errors. METHOD: Microscopic evaluation of cytospin preparations of the top layer of the plasma was compared with the difference between duplicates in the IFCC-recommended LDH assay using a Hitachi 911 analyser. RESULTS: Duplicate errors could not be explained by the presence of one of the individual cell types. In 16 samples with cellular aggregates absent, three did show duplicate errors; all three had a moderate amount of red blood cells present. CONCLUSION: The presence of cellular aggregates in the top layer of the plasma was the main determinant for the occurrence of the high frequency of duplicate errors in the IFCC-recommended LDH method. PMID- 17132286 TI - Assessment of the practicability and analytical performance of a point-of-care affinity chromatography haemoglobin A1c analyser for use in the non-laboratory setting. AB - BACKGROUND: Haemoglobin A(1c) (HbA(1c)) is a pivotal pathology test used around the world for the long-term management of patients with diabetes. Point-of-care testing (POCT) provides a convenient means for conducting HbA(1c) testing outside the laboratory. METHODS: The practicability and analytical performance of the Micromat II POCT HbA(1c) analyser (Bio-Rad Laboratories, USA), which has affinity chromatography as its methods principle, was evaluated in Australia and compared with the DCA 2000 POCT device (Bayer Australia) and a laboratory-based high performance liquid chromatography (HPLC) method. RESULTS: Overall between-day imprecision over 10 days was 1.9% for the laboratory HPLC method, 2.2% for the DCA 2000 and 7.0% for the Micromat II. In a second study over the same time period, the Micromat II's imprecision was 6.4%. The mean difference between the Micromat II and the laboratory method in a patient comparison (n = 100) was 0.25% (lower and upper limits of agreement -1.79 to 1.30). CONCLUSIONS: The imprecision obtained with the Micromat II was inferior to both the DCA 2000 and laboratory methods and did not meet current internationally accepted precision goals for this analyte. The Micromat II's poor imprecision can be explained by the high degree of technical expertise needed to perform the test; its use by non laboratory health professionals such as nurses and Aboriginal health workers in rural and remote Australia cannot be recommended. PMID- 17132287 TI - A mild case of abetalipoproteinaemia in association with subclinical hypothyroidism. AB - Abetalipoproteinaemia (ABL), an extremely rare recessive disorder, is characterized by exceptionally low or undetectable concentrations of apolipoprotein (apo) B-containing lipoproteins. ABL results from mutations in the gene encoding microsomal triglyceride transfer protein (MTP), a chaperone that facilitates the transfer of lipids onto apoB. Patients with ABL often present in childhood with a range of symptoms including fat malabsorption and manifestations of fat-soluble vitamin deficiencies. We describe a patient with sub-clinical hypothyroidism and ABL found to be compound heterozygous for a novel splice site mutation of intron 1 (c.61 + 2T > C) and a single adenine insertion in MTP exon 4 (c.419-420insA) that results in a frameshift and a protein truncated at 140 amino acids. PMID- 17132291 TI - The relationship between birthweight, 25-hydroxyvitamin D concentrations and bone mineral status in neonates. AB - BACKGROUND: Calcium (Ca), phosphorus (P) and 25-hydroxyvitamin D (25-OHD) are the major micronutrients for fetal skeletal development. AIMS: To compare whole body bone mineral density (WB BMD) and bone mineral content (WB BMC) in different birthweights of term neonates and to determine correlations of biological criteria of bone health between neonates and their mothers. SUBJECTS AND METHODS: Serum Ca, P, alkaline phosphatase (ALP) and 25-OHD levels were measured in 30 small-for-gestational-age (SGA, group 1), 40 appropriate-for-gestational-age (AGA, group 2) and 30 large-for-gestational-age (LGA, group 3) neonates and their mothers in winter. WB BMD and WB BMC of neonates were estimated by dual-energy X ray absorptiometry (DEXA) in the 1st 24 hrs after delivery. RESULTS: Mean (SD) serum 25-OHD levels in the mothers [8.7 (3.0), 8.6 (3.0) and 7.7 (2.8) microg/L, respectively] and their infants [6.3 (2.5), 6.0 (2.2) and 5.7 (1.8) microg/L, respectively] in groups 1, 2 and 3 were similar. Compared with the mothers, the mean 25-OHD levels of the neonates in all groups were significantly lower (p<0.05), and they were highly correlated (r=0.755, p<0.05). Ninety-three per cent of the neonates and 82% of their mothers had 25-OHD levels <10 microg/L, the lowest limit of normal. Mean (SD) WB BMD and WB BMC were higher in LGA infants [0.442 (0.025) g/cm(2), 71.6 (9.0) g, p<0.01, p<0.001, respectively] but lower in SGA [0.381 (0.027) g/cm(2), 29.1 (9.1) g, p<0.001, p<0.001, respectively] than in AGA infants [0.426 (0.022) g/cm(2), 53.7 (9.6) g, respectively]. The percentage of WB BMC was lower in SGA than in AGA and LGA infants. WB BMC and WB BMD were positively correlated with birthweight (r=0.910, p<0.05) and gestational age (r=0.707, p<0.05) but not with serum 25-OHD. CONCLUSIONS: The neonates' bone indices increased significantly with gestational age and birthweight but this was not related to serum 25-OHD levels in the infants and their mothers. PMID- 17132292 TI - Hypoxaemia among children in rural hospitals in Papua New Guinea: epidemiology and resource availability--a study to support a national oxygen programme. AB - AIMS: To support a national approach to oxygen systems in Papua New Guinea, we conducted a study to document the incidence of hypoxaemia, its geographical distribution, epidemiological determinants and resource availability in several regions of the country. We also established baseline mortality rate data for all children admitted to five hospitals, for children with a diagnosis of pneumonia and for neonates to evaluate a future intervention. METHODS: Data were collected prospectively from over 1300 hospital admissions in five hospitals in 2004. To establish the baseline case fatality rates, data on outcome were collected retrospectively over 3 years (2001-2003) for over 20,000 children admitted to five hospitals. RESULTS: A total of 1313 admissions were studied prospectively in the five hospitals. Altogether, 384 (29.25%, 95% CI 26.8-31.8) had hypoxaemia, defined as SpO(2) <90%. The incidence of hypoxaemia was much greater in highland hospitals (40% of all admissions) than on the coast (10% of all admissions). This large difference in incidence persisted when the uniform definition of hypoxaemia was adjusted for altitude, and was largely because of differences in the incidence of acute respiratory tract infection. Oxygen was not available on the day of admission for 22% of children (range between hospitals, 3-38), including 13% of all children with hypoxaemia. Oxygen was less available in remote rural district hospitals than in provincial hospitals in regional towns. Clinical signs proposed by WHO as indicators for oxygen would have missed 29% of children with hypoxaemia and, if these clinical signs were used, 30% of children without hypoxaemia would have been considered in need of supplemental oxygen. CONCLUSIONS: Based on this study, an approach to improving the detection of hypoxaemia and the availability of oxygen has been trialled in these five hospitals where a programme of clinical and technical training in the use and maintenance of pulse oximetry and oxygen concentrators has been introduced. PMID- 17132293 TI - Aetiological agents, interleukin-6, interleukin-8 and CRP concentrations in children with community- and hospital-acquired pneumonia. AB - OBJECTIVE: To determine the pathogens causing pneumonia in community-acquired pneumonia (CAP) and hospital-acquired pneumonia (HAP) and to investigate serum levels of interleukin-6 (IL-6), interleukin-8 (IL-8) and CRP in pneumonia caused by different aetiological agents. STUDY DESIGN: Eighty-seven children (mostly < 5 years of age) were recruited in a prospective study, 55 of them with CAP without prior antibiotic treatment and 32 with HAP. Thirty healthy outpatient children served as controls. RESULTS: The causative micro-organisms were determined by serological and microbiological methods in 40 cases with CAP (72.7%) and 30 with HAP (93.7%). In CAP, M. pneumoniae was the most common causative agent (43.6%), followed by S. pneumoniae (20%) and C. pneumoniae (18.1%). Bacteria alone were the sole causative agents in only 21.8% of cases with HAP. Pseudomonas aeruginosa (34.3%) and K. pneumoniae (32.5%) were the most frequently isolated. Although IL 6 and IL-8 levels were raised, there was no statistical difference between the CAP and HAP groups, or between bacterial and mycoplasma infections; neither was there a difference in CRP levels between these two groups. CONCLUSION: The causes of pneumonia differ between CAP and HAP. Levels of IL-6, IL-8 and CRP are raised in pneumonia but are unhelpful in differentiating the various aetiologies. PMID- 17132294 TI - Clinico-laboratory profile and outcome of Japanese encephalitis in Nepali children. AB - BACKGROUND: Japanese encephalitis (JE) is associated with high mortality and neurological sequelae. The unpredictable course and lack of specific treatment pose major challenges in management. The tropical climate and paddy ecosystem in Nepal provide a suitable setting. AIMS: To determine the aetiology of febrile encephalopathy and describe the clinico-laboratory profile and outcome of JE in Nepali children. METHODS: A hospital-based prospective and observational study was conducted over a 1-year period (2000-2001). Children aged from >1 month to 14 years with fever >38 degrees C for <2 weeks and altered sensorium were recruited. JE was confirmed by anti-JE IgM in cerebrospinal fluid and/or serum. RESULTS: Of 117 consecutively enrolled patients, 58 had JE. Ten patients had concomitant infections, four with malaria and six with bacterial meningitis, and were excluded from analysis. Clinical findings were as follows: boys, 69%; age 4-14 years, 71%; presentation during summer and autumn, 83%; fever >3 days, 69%; altered sensorium <2 days, 50%; Glasgow coma score 8-12, 63%; seizures, 58%. Four (8.3%) died. At discharge, neurological sequelae were detected in 24 (50%) and hemiparesis was the most common form. Longer duration of vomiting, altered sensorium and focal neurological deficit on admission were independently associated with sequelae at discharge. Sequelae persisted in nine (18.8%) at 6 weeks follow-up. Long duration of altered sensorium (beta co-efficient 0.35, odds ratio 1.4, p=0.042) and presence of focal neurological deficit on admission (beta co-efficient 1.6, odds ratio 5.2, p=0.049) were independent predictors of sequelae at 6 weeks. CONCLUSION: JE was the commonest cause of febrile encephalopathy. Neurological sequelae were common but resolved in two-thirds of cases. PMID- 17132295 TI - Prevalence and determinants of the practice of genital mutilation of girls in Khartoum, Sudan. AB - BACKGROUND: Female genital mutilation (FGM) is widely practised in Sudan, despite many decades of attempts to prevent it. AIMS: To estimate the prevalence of FGM, identify the types performed in Khartoum and investigate whether FGM is associated with various social factors. METHODS: Girls aged 4-9 years (n=255) presenting to a paediatric emergency ward were recruited. A detailed history was obtained and full examination, including inspection of the genitalia, was performed to verify the type and extent of FGM. RESULTS: Twenty per cent of the study group had undergone FGM, 50% of guardians indicated that it would be done later, and 29% stated that the child would not undergo FGM. In 66% of those who had undergone FGM, it was WHO type III. All operations had been performed by health professionals, mainly midwives. Those who had allowed or intended to allow their daughters to undergo FGM were of significantly lower socio-economic status (p=0.0008) and had spent significantly fewer years in school (both mothers, p=0.0015, and fathers, p=0.0266) than those who had not/would not. All who had undergone FGM were Muslims. None of the 16 Christians had undergone FGM. In girls over 7 years of age, there was a higher risk of having FGM in those who attended school than in those who did not. CONCLUSION: FGM is still practised widely in Khartoum and probably in many parts of northern Sudan and the type undertaken is often the most severe. Parental education, socio-economic level and religion are important determinants of the practice, but social pressure on parents and girls seems to play an important role. PMID- 17132296 TI - Significance of coagulase-negative staphylococci isolated from a single blood culture from neonates in intensive care. AB - BACKGROUND: Coagulase-negative staphylococci (CoNS) are the most common pathogens of late-onset bacteraemia in neonatal intensive care units (NICUs). Discriminating true infection from contamination is difficult. METHODS: To evaluate the significance of a single blood culture yielding CoNS from only one blood culture obtained from NICU infants between July 1999 and November 2000, paired blood cultures were obtained simultaneously from two peripheral sites from infants hospitalised in two NICUs with clinically suspected late-onset sepsis, and a single blood culture was obtained peripherally from infants hospitalised in a third NICU. Demographic data, predisposing factors, clinical manifestations, laboratory data, management and outcome of infants with either paired or single blood cultures yielding CoNS were analysed and compared. RESULTS: Both blood cultures yielded CoNS during 26 episodes in 24 infants. A single blood culture which grew CoNS from one blood culture was identified during 24 episodes in 23 infants. Except for prior use of an umbilical venous catheter, there was no significant difference between the infants with CoNS isolated from a paired or a single blood culture in terms of demographic characteristics, predisposing factors, clinical manifestations, laboratory findings, management and outcome. CONCLUSION: There was no difference in the clinical features of CoNS infection between single and paired CoNS-positive blood cultures. PMID- 17132297 TI - Bacteraemia in severely malnourished children in an HIV-endemic setting. AB - BACKGROUND: HIV infection predisposes children with malnutrition to recurrent bacterial infections and a high risk of bacteraemia. METHODS: A cross-sectional descriptive study to determine the prevalence, causative organisms, antibiotic sensitivity and factors associated with bacteraemia in malnourished children was undertaken at Mulago Hospital, Kampala. The prevalence of HIV infection was also determined. A total of 134 children aged 6-59 months with severe malnutrition were recruited. RESULTS: Sixty-one (45.5%) had oedematous malnutrition and 73 (54.5%) had severe wasting. Fifty-nine (44.0%) were HIV-infected. The prevalence of bacteraemia was 22%. The predominant organisms isolated were gram-negative enteric bacilli (77%) with Salmonella species and E. coli contributing 67% of the isolates. Hypoglycaemia was significantly associated with bacteraemia (p=0.007). Most organisms were resistant to cotrimaxazole (93.3%), ampicillin (76.7%), gentamicin (66.7%) and chloramphenicol (60%). All isolates were sensitive to ceftriaxone. Sensitivity to ciprofloxacin was 97%. There was no strong association between HIV infection and bacteraemia. The relative risk of death in malnourished children with bacteraemia was ten times higher than in those without bacteraemia. CONCLUSIONS: Nearly a quarter (22%) of children admitted with severe malnutrition had bacteraemia and gram-negative organisms were the predominant cause. Forty-four per cent were HIV-infected. Most of the bacteria were sensitive to ceftriaxone and ciprofloxacin and resistant to commonly used antibiotics. In the absence of culture and sensitivity, ciprofloxacin or ceftriaxone should be considered as first-line antibiotics for severely malnourished children. PMID- 17132298 TI - Detection of antigenuria for diagnosis of invasive Haemophilus influenzae type b disease. AB - BACKGROUND: Haemophilus influenzae type b (Hib) diseases are responsible for an estimated 400,000 childhood deaths, mostly in developing countries. OBJECTIVES: To determine the value of the Wellcogen quantitative latex agglutination test (LA) in urine for the diagnosis of Hib pneumonia and meningitis. METHODS: Healthy and sick children aged <5 y were enrolled in Dhaka Shishu (Children's) Hospital. Boiled and non-concentrated urine specimens underwent LA testing. In vaccinated subjects, urine was tested by LA at 24 h, 4-6 and 7-10 d after vaccination. RESULTS: Of 1302 enrolled cases, 201 were healthy (90 Hib vaccine recipients and 111 provided NP) and 1101 were sick with either pneumonia (n=974) or meningitis (n=127). Among the healthy children enrolled, 41 (41/111, 37%) were colonised with Hib and two (2/41, 5%) were positive by LA test. Hib antigenuria among the children who had received Hib vaccination was mainly detected only on day 1 (7/90, 8%) of vaccination. Among the sick children, LA test for Hib antigen was positive for all confirmed cases of Hib pneumonia (10) and meningitis (35). In contrast, none of the urine specimens from the cases with a known aetiology other than Hib (n=104) was positive. Quantitative analysis of antigenuria of sick children showed that it is positive at least up to 1:8 and 1:16 dilutions for pneumonia and meningitis, respectively, in contrast with or=1:8 dilutions. CONCLUSIONS: The Wellcogen LA test for Hib using boiled and non-concentrated urine is more sensitive than blood culture alone and is highly specific. PMID- 17132299 TI - Rectal bleeding in Egyptian children. AB - AIM: In a prospective study to outline the aetiology of bleeding per rectum (BPR) in Egyptian infants and children, a subsidiary aim was to define some of the clinical characteristics of the different aetiologies. SUBJECTS AND METHODS: 194 children with BPR are described. The diagnostic work-up included laboratory investigations, radiological and endoscopic assessment, radio-isotope scanning, angiography and histopathological examination of mucosal biopsies, as appropriate. RESULTS: Ages ranged from 3 to 192 months with a mean (SD) of 49.8 (43.5). Infectious enterocolitis was the most common cause (37.1%). Others included colorectal polyps (21.1%), chronic colitis (16%) including inflammatory bowel diseases (5.2%), allergic colitis (2.6%), solitary rectal ulcer syndrome (1.5%) and non-specific colitis (6.7%). Intussusception and Meckel's diverticulae were the cause in 7.3% and 2.6%, respectively, while other aetiologies included vascular (6.2%), systemic (3.6%), local anal (3.1%) and upper gastro-intestinal causes (1.5%). In 1.5% of cases, the cause remained 'obscure'. CONCLUSION: In Egyptian children, infectious enterocolitis followed by colorectal polyps and chronic colitis are major causes of BPR. PMID- 17132300 TI - Incidental raised transaminases: a clue to muscle disease. AB - Twenty-one patients with incidental hypertransaminasaemia who were eventually diagnosed as muscular dystrophy are described. There were two females and 19 males aged between 2 and 11 years [mean (SD) 6.7 (3.4) y]. Serum alanine and aspartate transaminase levels were between 73 and 595 IU/L (30-35) and 68 and 550 IU/L (30-35), respectively. Muscle disease was suspected when creatine phosphokinase levels were elevated and confirmed in each patient by muscle biopsy. The time interval between incidental hypertransaminasemia and the diagnosis of muscle disease was between 3 and 12 months. Eleven patients were diagnosed as Becker's muscle dystrophy, eight as Duchenne muscle dystrophy and two had sarcoglycanopathy. Long-term elevation of transaminase levels might be a sign of occult muscle disease. Invasive tests such as liver biopsy should not be performed in patients with unexplained hypertransaminasaemia without first determining creatinine phosphokinase levels. PMID- 17132301 TI - Childhood rhabdomyosarcoma in Ibadan, Nigeria: 1984-2003. AB - BACKGROUND: Rhabdomyosarcoma is the most common soft tissue sarcoma in children under 15 years of age. There is, however, a paucity of reports on the pattern of its occurrence in Nigeria and other parts of Africa. AIM: To describe the pattern of childhood rhabdomyosarcoma in Ibadan with respect to demography, morphology and tumour site. METHODS: This retrospective study was based on data obtained from the Ibadan cancer registry, College of Medicine, University of Ibadan. All histologically confirmed cases of rhabdomyosarcoma in children under 15 years of age seen at the University College Hospital Ibadan between 1984 and 2003 were included. Information obtained includes age, gender, morphology and sites of the tumours. RESULTS: Ninety-one children were seen with a male:female ratio of 1.5:1. Ages ranged from <1 to 14 years with a mean (SD) of 6.2 (4.1) years. The embryonal variant was the most common histological subtype (61.5%). Other subtypes were alveolar (13.2%), pleomorphic (4.4%) and rhabdomyosarcoma 'not otherwise specified' (20.9%). The majority (50.6%) of tumours were in the head and neck region and the common primary sites were soft tissue of the head, face (24.2%) and orbit (14.3%). Other sites included soft tissue of the pelvis (11.0%), genito-urinary tract (9.9%) and abdomen (9.9%). CONCLUSION: The pattern of rhabdomyosarcoma in Nigeria is similar to that in the United States and Europe, except for the rarity of parameningeal sites and extremities. There is a need for larger descriptive studies on childhood rhabdomyosarcoma in Africa. PMID- 17132302 TI - Safety and immunogenicity of a candidate vaccine for visceral leishmaniasis (Alum precipitated autoclaved Leishmania major + BCG) in children: an extended phase II study. AB - BACKGROUND: Untreated visceral leishmaniasis (VL) is an inevitably fatal childhood disease. First-generation candidate vaccines for VL [autoclaved Leishmania major (ALM) + BCG] have been found to be safe and immunogenic but not superior to BCG alone. Modulation of ALM by adsorption to Alum significantly increases the immunogenicity. The Alum-adsorbed ALM vaccine was found to be safe and strongly immunogenic in healthy adult volunteers in a non-VL-endemic area. This study aimed at establishing the safety and immunogenicity of Alum precipitated autoclaved L. major + BCG vaccine in children under field conditions. METHODS: A total of 544 healthy, leishmanin non-reactive children (<15 y) were randomly allocated to receive either a single intradermal injection of Alum/ALM + BCG or vaccine diluent (placebo). Volunteers were closely followed for 2 years at 6-month intervals for vaccine safety and immunogenicity. RESULTS: The vaccine was well tolerated with minimal side-effects. Leishmanin skin test conversion (>or=5 mm) was seen in 56%, 50%, 25% and 31% at 6, 12, 18 and 24 months post-vaccination, respectively; conversion in the placebo group was 4%, 12%, 3% and 13% at the same follow-up visits. There was no significant increase in anti-leishmanial antibodies in either study arm at any of the follow-up visits. During the study, four patients in the placebo arm developed parasitologically confirmed VL. CONCLUSION: Alum/ALM + BCG vaccine is safe and immunogenic in children under field conditions. Multiple injections might be needed to obtain results similar to those obtained in healthy volunteers. PMID- 17132303 TI - Acute muscle weakness owing to severe hypokalaemia resulting from dietary insufficiency: case report. AB - Acute muscle weakness with severe hypokalaemia is not uncommon in adults but is rare in children. An 11-month-old girl presented with hypokalaemic paralysis following a 1-month insufficiency of dietary potassium. PMID- 17132304 TI - Brucellosis with pulmonary involvement in a premature infant. AB - Congenital brucellosis is rare. A premature infant with transplacentally acquired congenital brucellosis and pulmonary involvement is described. PMID- 17132305 TI - Meconium peritonitis and parvovirus B19 infection associated with hydrops fetalis. AB - Two preterm infants with non-immune hydrops fetalis associated with meconium peritonitis are reported. The first presented with a cystic abdominal mass and the second had positive parvovirus B19 serology. The association of meconium peritonitis with hydrops fetalis was through different mechanisms in each patient. PMID- 17132306 TI - The current standard of care in the periprocedural management of the patient with obstructive jaundice. AB - This review provides a literature-based guide to the optimal management of the patient with obstructive jaundice with emphasis placed on prevention of complications. PMID- 17132307 TI - Sacral nerve neuromodulation for the treatment of lower bowel motility disorders. AB - INTRODUCTION: Incontinence and constipation are common and cause a high degree of physical, social and psychological impairment. Maximal conservative therapy may improve some patients but many remain symptomatic. Surgical options are often unsatisfactory, with variable result and further options are limited. Sacral nerve stimulation uses electrical stimulation applied to the sacral nerves, eliciting a physiological effect on the lower bowel, anal sphincter and pelvic floor, resulting in clinical benefit. The objective of this study was to investigate whether sacral nerve neuromodulation can improve patients with disorders of bowel motility, when current maximal treatment has failed and to investigate the underlying physiological mechanism of action. RESULTS: Incontinence: Nineteen patients, age 58 years (range, 37-71 years), with resistant incontinence for 6 years (range, 2-21 years) underwent stimulation. Continence improved in all at 24 months (range, 3-60 months), fourteen fully continent. Incontinent episodes decreased; 12 (range, 2-30) versus 0 (range, 0 4), P < 0.001. Urgency (P < 0.01) and quality of life improved (P < 0.05). Anal squeeze pressure (P = 0.001) and rectal sensation (P < 0.01) improved. Constipation: Four women, (aged 27-36 years) with resistant idiopathic constipation for 8-32 years underwent the first worldwide implants. Symptoms improved in all with temporary, and in three with permanent, stimulation at 8 months (range, 1-11 months). Bowel frequency increased: 1-5 versus 6-28 evacuations/3-weeks. Symptom scores and quality of life improved. Placebo effect: A double-blind, cross-over study was performed to examine placebo effect and efficacy. Once stimulation was removed, in a blinded manner, symptoms, physiological parameters and quality of life measures rapidly returned to baseline levels. Autonomic neuromodulation: Sixteen patients, median age 59 years (range, 38-71 years), were studied at 27 months (range, 2-62 years) using laser Doppler flowmetry. Chronic stimulation was at 2.8 V (range, 0.3-3.9 V). Median flux differed between none and chronic stimulation (P = 0.001). Step-wise increments caused an immediate, dose-dependent rise in flux (P < 0.0001) up to 1.0 V. CONCLUSIONS: This research provides strong evidence that sacral nerve stimulation can improve patients with resistant incontinence and shows proof-of concept for the treatment of constipation. The effect is unlikely to be due to placebo and the mechanism is rapidly reversible and involves a dose-dependent effect on the autonomic nerves. PMID- 17132309 TI - Corress feedback. PMID- 17132308 TI - Aggressive management of surgical emergencies. AB - Increasing evidence suggests that two factors significantly influence outcome in a surgical emergency - premorbid health and the degree of inflammation during the first 24 h following trauma. Repeat observations suggest that the depth of post trauma immunoparalysis reflects the height of early inflammatory response. Administration to surgical emergencies, as was routine in the past, of larger amounts of fluid and electrolytes, fat, sugar and nutrients seems counterproductive as it increases immune dysfunction, impairs resistance to disease and, in fact, increases morbidity. Instead, strong efforts should be made to limit the obvious superinflammation, which occurs during the first 24 h after trauma and, thereby, reduce the subsequent immunoparalysis. Several approaches show efficacy in limiting early superinflammation such as strict control of blood glucose, avoidance of stored blood when possible, supply of antioxidants, live lactic acid bacteria and plant fibres. This review focuses mainly on use of live lactic acid bacteria and plant fibres, often called synbiotics. Encouraging experience is reported from clinical trials in liver transplantation, severe pancreatitis and extensive trauma. Immediate control of inflammation by enteral nutrition and supply of antioxidants, lactic acid bacteria and fibres is facilitated by feeding tubes, introduced as early as possible on arrival at the hospital. PMID- 17132310 TI - Lymphoscintigraphy in detection of the regional lymph node involvement in gastric cancer. AB - INTRODUCTION: Involvement of regional lymph node is a critical sign in prognosis of gastric cancer. Radiological techniques are commonly used to evaluate the extension of gastric cancer. But their sensitivity and specificity are low especially in the early stage. Our aim was to assess the value of gastric lymphoscintigraphy in identifying regional lymph node involvement in patients with gastric cancer, as compared to the abdominal ultrasonography, computed tomography and postoperative histopathological evaluation. PATIENTS AND METHODS: 50 patients (12 females) with a median age of 61 years (range, 35-73 years) were included in the study. Pre-operative staging in all cases included upper gastrointestinal endoscopy and biopsy, followed by ultrasound, computed tomography and lymphoscintigraphy. 148 MBq Technetium-99m lymphoscint was injected around the tumour during endoscopy and immediately after injection, anterior, lateral and posterior images were taken in 5-min intervals using a gamma camera. Findings were compared to the findings of other tests. The sensitivity, specificity, positive predictive value, and negative predictive value of each test were calculated and compared. RESULTS: Histologically, 68% of cases (34/50) had metastasis in regional lymph nodes and all cases were accurately diagnosed by lymphoscintigraphy. Lymphoscintigraphy was significantly more sensitive for detecting lymph node involvement (P < 0.01). Both abdominal ultrasonography and CT had very low sensitivity in identifying lymph nodes. CONCLUSIONS: Lymphoscintigraphy is a promising test in the identification of regional lymph nodes pre-operatively in patients with gastric cancer. It might help the surgeon to plan the extent of dissection before surgery which may decrease postoperative complications related to unnecessary extensive lymph node dissection. PMID- 17132311 TI - Radiofrequency ablation (RFA) as a cytoreductive strategy for hepatic metastasis from breast cancer. AB - INTRODUCTION: Patients with liver metastasis from breast cancer have a poor prognosis, although this may be improved by hepatectomy in a selected group with disease confined to the liver. We evaluate the effectiveness of radiofrequency ablation (RFA) as a cytoreductive strategy in the management of liver metastasis from primary breast cancer. PATIENTS AND METHODS: Nineteen patients with hepatic metastasis from primary breast cancer underwent RFA of their liver lesions between April 1998 and August 2004. RESULTS: The median age of the patients was 52 years (range, 32-69 years), 8 had disease confined to the liver, with 11 having stable extrahepatic disease in addition. Seven patients with disease confined to the liver at presentation are alive, as are 6 with extrahepatic disease, median follow-up after RFA was 15 months (range, 0-77 months). Survival at 30 months was 41.6%. In addition, 7 patients followed up for a median of 14 months (range, 2-29 months) remain alive and disease-free. RFA failed to control hepatic disease in 3 patients. RFA was not associated with any mortality or major morbidity. CONCLUSIONS: Control of hepatic metastasis from breast cancer is possible using RFA and may lead to a survival benefit, particularly in those patients with disease confined to the liver. PMID- 17132312 TI - Should we apply suction during fine needle cytology of thyroid lesions? A systematic review and meta-analysis. AB - INTRODUCTION: Fine needle aspiration cytology (FNAC) is a well-established investigation in thyroid disease. Fine needle sampling without aspiration (FNS) is less commonly used but often easier to perform. Both methods have advantages and disadvantages but, as yet, there is no agreement on which method produces better specimens for cytological diagnosis. MATERIALS AND METHODS: We undertook a review of the literature and performed a meta-analysis of the results of four cross-over trials. RESULTS: The resulting odds ratio favoured FNS (OR = 0.99; 95% CI 0.88-1.11) but was not statistically significant. A fifth paper not included in the meta-analysis reported results in favour of FNS (P = 0.003). CONCLUSIONS: There is no evidence from the meta-analysis that one method is superior to the other; however, taking into consideration all available evidence, it seems that FNS may be easier to perform and may produce better samples. PMID- 17132313 TI - Vascular surgical emergencies: how will future surgeons be trained? AB - INTRODUCTION: We wished to assess whether pattern and impact of emergency vascular surgical referrals has altered since a previous study in 1990. Following introduction of shift working patterns, we wished to assess how these changes may affect vascular training and vascular on-call cover. PATIENTS AND METHODS: Prospective survey of emergency vascular referrals at two district general hospitals (DGH-R and DGH-L) in 2003. DGH-R received only regional referrals whereas DGH-L also received 'next day' referrals from a smaller hospital. Results were compared between centres and with a previous study undertaken at DGH-R in 1990. RESULTS: From 1990 to 2003 emergency vascular referrals at DGH-R increased by 51% (53 to 80). The number seen at DGH-R and DGH-L were similar in 2003. There were significantly more out-of-hours referrals in DGH-R than DGH-L (59% versus 35%; P = 0.0123). Referrals were more likely to be seen initially by the vascular team at DGH-L than DGH-R (80% versus 47%, P < 0.0001). CONCLUSIONS: Vascular emergency referrals have increased. A trainee was likely to see more emergency referrals at DGH-L than DGH-R. This may impact on future training. PMID- 17132314 TI - A waiting list to go home: an analysis of delayed discharges from surgical beds. AB - INTRODUCTION: The aim of this paper was to analyse patients who were unable to be discharged from a surgical ward despite being surgically fit to leave. PATIENTS AND METHODS: Data were collected on all surgical in-patients on a single day. Patients who were surgically fit for discharge but whose discharge was delayed were identified. Demographic data and reasons for delay were noted. RESULTS: Nine of 75 patients (12%) were surgical bed blockers. These patients were more likely to have been admitted as emergencies (P = 0.035) and were older (P < 0.01) than the remaining patients. They occupied 35% of the total 'bed-days' of the group as a whole with a median in-patient stay of 41 days compared with 2 days for the other patients. Trust-collected data, based on UK Government guidelines, showed only one surgical delayed discharge patient on the day studied. CONCLUSIONS: Due to problems in defining delayed discharge Government figures probably underestimate the true numbers. Lack of intermediate care and social service provision are a major cause of bed blocking. PMID- 17132315 TI - The extended scope physiotherapist in orthopaedic out-patients - an audit. AB - INTRODUCTION: We undertook an audit of the activity of the extended scope physiotherapists (ESPs) in our unit. We assessed their activity against three benchmark data: (i) independent assessment and management by the ESP of 85% of patients seen by them; (ii) no patient to be re-referred to a surgeon with the same problem; and (iii) patient satisfaction rate of 89%. PATIENTS AND METHODS: All new referrals seen by the ESPs over the 6-month period between July 2002 and December 2002 were included in the audit. Patient medical records were reviewed retrospectively. Patients were contacted by telephone 12 months after their initial clinic appointment to obtain patient satisfaction scores. RESULTS: In the 6-month period, 150 patients (75 male, 75 female) were seen. Their median age was 43.5 years (range, 17-85 years). Their main complaints related to the spine (42%), knee (33%), shoulder (18%), or other site (7%). The ESPs saw and managed 82/150 patients (55%) independently. Consultant review was required for 81% of shoulder cases, 34% of knee cases and 11% of back cases. GPs re-referred 4/150 patients (3%). We successfully contacted 126 patients by telephone: 97 (77%) were satisfied with their management by ESPs. Of patients who were dissatisfied, 76% did not see a consultant at any stage in their management. CONCLUSIONS: ESPs fulfilled a useful role in our orthopaedic out-patient clinic particularly in the back clinic. The percentage of independently managed patients was much lower than the figure quoted in the literature. We suspect that the published literature does not reflect modern referral patterns and recommend that time for review of new patients seen by ESPs should be factored into consultant's clinic time. Patients' expectations may be a barrier to achieving greater levels of patient satisfaction. PMID- 17132316 TI - Day-care surgery for pilonidal sinus. AB - INTRODUCTION: The surgical approach to symptomatic pilonidal sinus is open to debate. Many techniques have been described and no single technique fulfils all the requirements of an ideal treatment. Ambulatory treatment with minimal morbidity and rapid return to activity is desirable. The aim of this work was to study the feasibility of day-care surgery for excision and primary asymmetric closure of symptomatic pilonidal sinus. PATIENTS AND METHODS: All patients referred electively over 2 years were assessed in a single-consultant, colorectal clinic and booked for day-care surgery. All patients had excision and primary asymmetric closure under general anaesthesia in the left lateral position. Whenever possible, they were discharged on the same day according to the day surgery protocol. Patients were subsequently seen in the out-patient clinic for removal of stitches and were followed up further if there was any wound breakdown. RESULTS: Fifty-one patients were operated on electively for pilonidal sinus over the 2 years. Two patients were excluded as the final diagnosis was not pilonidal sinus. At 4 weeks following operation, 43 (88%) had complete healing and 6 (12%) had dehiscence of the wound. Recurrence rate was 8% (4 patients) for follow-up of 12-38 months. There was no admission from the day-surgery unit and no unplanned re-admissions. The cost for day-care pilonidal sinus surgery was estimated to be 672.00 pounds per patient compared with in-patient cost of 2405.00 pounds. CONCLUSIONS: Excision and primary asymmetric closure for pilonidal sinus is safe and feasible as day-care surgery and is associated with potential cost saving. PMID- 17132317 TI - Interpreter services in an inner city teaching hospital: a 6-year experience. AB - INTRODUCTION: Being able to communicate effectively with patients is essential not only from a medicolegal standpoint but more importantly from clinical governance perspectives. Issues such as informed consent and patient choice within the NHS are currently being highlighted; for these to be available to patients, their language requirements are paramount. PATIENTS AND METHODS: An audit was performed by the Linkworkers office at the Central Manchester & Manchester Children's Hospital NHS (CMMC) Trust on the total number of attendances and refusals per language in the period 1998-2003. RESULTS: In the CMMC Trust, Urdu/Punjabi, Bengali, Cantonese, Somali, Arabic and French represent the majority of the workload, comprising almost 80% of cases in 2003. In the same year, an increase in demand for languages of Eastern European countries became evident. Finding interpreters for these languages even via agencies can be extremely difficult. CONCLUSIONS: If the current trend continues, requirement for these services will increase exponentially. For this demand to be met adequately these issues must be kept at the forefront of NHS planning. PMID- 17132318 TI - Broken bones and fractures - an audit of patients' perceptions. AB - INTRODUCTION: 'Have I got a fracture or a break doctor?' remains a commonly posed question in fracture clinics, suggesting that patients frequently feel a 'fracture' and a 'break' are two separate entities. This apparent misconception amongst fracture clinic patients may result in confusion and occasionally anger that doctors appear to have inconsistent views on the severity of their injury. Compliance and outcome of patient care can also be affected by poor communication. PATIENTS AND METHODS: Our questionnaire-based study was conducted in two stages. The initial objective was to establish whether this misconception surrounding the words 'fracture' and 'break' is commonly held amongst our out patient trauma patients. The second stage of the audit was to determine whether a patient information leaflet on fractures/broken bones could help reduce this misconception. RESULTS: The preliminary audit involving 50 new patients attending our fracture clinic showed that 84% thought there was a difference between a 'fracture' and a 'break', with 68% believing a 'break' to be worse than a 'fracture'. Following the introduction of an information leaflet, a re-audit of 61 new patients took place. This time 67% felt there was a difference between a 'fracture' and a 'break', with 65% believing a 'break' to be worse than a 'fracture'. Only 21% had read the supplied information leaflet, and 69% of those still believed there was a difference between a 'fracture' and a 'break'. CONCLUSIONS: The majority of patients believed that there was a difference between a 'fracture' and a 'break'. Access to information leaflets did not appear to alter this misconception. Verbal communication and explanation may be more beneficial and practical than visual aids and leaflets in overcoming this problem. PMID- 17132319 TI - Impact of changing trend in cold ischaemic time on operating times in renal transplantation. AB - INTRODUCTION: The European Working Time Directive (EWTD) is calling for reduction in the working hours of doctors. Renal transplantation is well-recognised as an out-of-hours specialty. Our study looks at whether our renal transplant centre's attempt to reduce cold ischaemic time (CIT) has impacted on the pattern of operating times since this may have implications on the surgeons' working hours. PATIENTS AND METHODS: We studied 883 adult cadaver kidney transplants performed between 1 January 1992 and 31 December 2002. CIT and time of surgery was obtained from a local audit database () and that held by UK Transplant. Time of surgery was divided into 07:30-17:59 (day),18:00-23:59 (evening) and 00:00-07:29 (night). RESULTS: CIT has decreased since 1992, with a significant reduction after 1998 (P = 0.0001). There was, however, no difference in the percentage of operations performed during the three time periods before and after 1998. Between 1999-2002, 51.4% of transplants took place out of hours (i.e. 18:00 07:29). CONCLUSIONS: Reduction in CIT does not appear to have impacted on our operating times. The large amount of out-of-hours work is, however, not compliant with EWTD requirements. PMID- 17132320 TI - Inflamed solitary caecal diverticulum - it is not appendicitis, what should I do? AB - We describe three cases that presented with symptoms suggestive of appendicitis but were found at operation to have an inflamed solitary caecal diverticulum. All were treated successfully with diverticulectomy or inversion of the diverticulum. We wish to highlight this diagnosis and its surgical management so that informed decisions can be made if this is first encountered in the operating theatre. PMID- 17132321 TI - Methicillin-resistant Staphylococcus aureus infection following arthroscopy of the knee joint. AB - Arthroscopic surgery of the knee is considered to be a safe procedure. We had a microbiologically confirmed infection of methicillin-resistant Staphylococcus aureus (MRSA). Although various rare infective cases are reported following arthroscopy of the knee joint, to the best of our knowledge there is no previous report of MRSA infection following arthroscopy of the knee joint. PMID- 17132324 TI - Perineural fibrosis of the paravasal nerves: a possible cause of orchalgia. PMID- 17132325 TI - A surgeon's or an athlete's foot. PMID- 17132327 TI - A better way to insert urethral catheter with guide-wire. PMID- 17132329 TI - Ethical Aspects of Avian Influenza Pandemic Preparedness. Proceedings of a symposium, 2005. PMID- 17132330 TI - Ethical issues in international vaccine research and development. PMID- 17132331 TI - An ethics framework for public health and avian influenza pandemic preparedness. PMID- 17132332 TI - The United States vaccine supply: challenges in preparing for an avian influenza pandemic. PMID- 17132333 TI - Monitoring vaccine safety during an influenza pandemic. AB - In the event that a vaccine is available during an influenza pandemic, vaccine safety monitoring will occur as part of comprehensive public health surveillance of the vaccination campaign. Though inactivated influenza vaccines have been widely used in the United States and much is known about their safety profile, attention will need to be paid to both common self-limited adverse reactions and rarer, more serious events that may or may not be causally related to vaccination. The primary surveillance systems used to generate and test hypotheses about vaccine safety concerns are the Vaccine Adverse Event Reporting System (VAERS) and the Vaccine Safety Datalink (VSD), respectively. Examples of recent use of these systems to investigate influenza vaccine safety and enhancements planned for use during a pandemic are presented. Ethical issues that will need to be addressed as part of an overall vaccine safety response include risk communication and injury compensation. Advance planning and the use of available technologic solutions are needed to respond to the scientific and logistic challenges involved in safely implementing mass vaccination during a pandemic. PMID- 17132334 TI - Public health strategies for distribution of influenza vaccine during an influenza pandemic. AB - In order to consider the ethical issues around vaccine distribution during an influenza pandemic, it is critical to have an understanding of the role of influenza vaccine in a pandemic, the rate at which vaccine is likely to be come available, who will likely produce and "own" the vaccine, how vaccine distribution and administration might be accomplished, and which are the groups that might be deemed highest priority to be vaccinated against influenza. The United States and Connecticut have been considering the more challenging of these issues and have learned from Canada, which previously discussed and made decisions on the challenges related to vaccine distribution. Although there is still some critical advance thinking that needs to be done, planning for the response to an influenza pandemic is now at an advanced stage. The keys to preparedness at this stage are to be aware of the vaccine distribution options, to know the benefits and limitations of each option, and to be flexible but nimble in dealing with a real pandemic. PMID- 17132335 TI - Rationing vaccine during an avian influenza pandemic: why it won't be easy. PMID- 17132336 TI - What about the ducks? An alternative vaccination strategy. AB - Like most emerging disease threats, avian influenza is a zoonotic disease maintained in nature by wildlife. In this case, the reservoir of infection is migratory waterfowl, primarily ducks. Rather than trying to vaccinate most of the world's human population in response to the threat of an avian influenza pandemic, it might be more prudent to vaccinate key reservoir wildlife species from which pandemic strains evolve. This strategy would require a much more intensive research effort to understand the evolution of avian influenza viruses in nature, but it would be far less costly than any of the alternatives. Research priorities for emerging zoonoses, such as new strains of avian influenza viruses, should be re-evaluated with an emphasis on ways to intervene at their source, the natural reservoir hosts from which they originate, rather than focusing up on human-based interventions, which are too often too late. PMID- 17132337 TI - Panglobalism and pandemics: ecological and ethical concerns. AB - A pandemic is a human medical problem but must be understood at multiple levels. Analysis of social and commercial forces is vital, and, more comprehensively, an ecological framework is necessary for an inclusive picture. Ecological health webworked with political and social determinants surrounds issues of human health. In this constellation of both natural and social factors, ethical concerns will arise at these multiple levels, from human health to the conservation and health of wild nature. PMID- 17132338 TI - Pandemic influenza: overview of vaccines and antiviral drugs. AB - Pandemic influenza has become a high priority item for all public health authorities. An influenza pandemic is believed to be imminent, and scientists agree that it will be a matter of when, where, and what will be the causative agent. Recently, most attention has been directed to human cases of avian influenza caused by a H5N1 avian influenza virus. An effective vaccine will be needed to substantially reduce the impact of an influenza pandemic. Current influenza vaccine manufacturing technology is not adequate to support vaccine production in the event of an avian influenza outbreak, and it has now become clear that new innovative production technology is required. Antiviral drugs, on the other hand, can play a very important role in slowing the disease spread but are in short supply and resistance has been a major issue. Here, we provide an update on the status of pandemic vaccine development and antiviral drugs. Finally, we conclude with some proposed areas of focus in pandemic vaccine preparedness. PMID- 17132339 TI - Public health and ethical considerations in planning for quarantine. PMID- 17132340 TI - Enhancing the legitimacy of public health response in pandemic influenza planning: lessons from SARS. PMID- 17132341 TI - Emerging diseases at the interface of people, domestic animals and wildlife. The role of wildlife in our understanding of highly pathogenic avian influenza. PMID- 17132342 TI - The next pandemic: anticipating an overwhelmed health care system. AB - INTRODUCTION: In September 2005, an overview of current health care system planning efforts was presented to the audience at the Yale University Ethics Symposium on Avian and Pandemic Influenza. The speaker, also the author of this article, provided the audience with a summary of what was being undertaken with the use of federal preparedness funds to improve the overall infrastructure of the health care system. All of Connecticut's 31 acute care hospitals, the Veteran's Administration Hospital in West Haven, Hospital for Special Care, Gaylord Rehabilitation Hospital, Natchaug Psychiatric Hospital, and the state's 13 Community Health Centers are currently recipients of federal preparedness funds. Federal funding for this planning comes from Health Resources and Services Administration, Department of Health and Human Service's National Bioterrorism Hospital Preparedness Program. OBJECTIVES: This article outlines the planning activities around pandemic influenza that the state's health care system partners started in 2004-2005 and also those they are currently participating in or will be participating in the next 12 to 15 months. The article highlights the key objectives and strategies that health care facilities will be using in this planning. There are four major objectives that each health care facility's Emergency Operations Plan must address. They are: increasing bed availability, developing strategies to deal with the potential staffing shortages, developing strategies for dealing with potential critical equipment and pharmaceutical shortages, and, lastly, the implementation of education, training and communication strategies for their health care workers and the public they serve. These plans, and all the activities needed to operationalize the plans, such as education, training, drills, and exercises, will include their key partners, i.e., local health departments, local emergency management, police, fire, and Emergency Medical Services. This article will describe this work plan in detail. METHODS: Descriptive information was obtained through the author's observations and personal experiences, in addition to governmental guidance, reports, and plans. CONCLUSION: The "all-hazards" planning currently being undertaken by the key health care system partners in Connecticut as a result of federal funding for preparedness post 9/11 has fostered great working relationships between these entities and their local, regional, and statewide planning counterparts. Many of the specific grant dollars being provided to these facilities can assist in the planning that must be done for pandemic flu. PMID- 17132343 TI - Of epidemic proportions: physicians, personal risk, and public trust. PMID- 17132344 TI - Talking to the public about a pandemic: some applications of the WHO outbreak communication guidelines. PMID- 17132347 TI - Changing disease incidence: the consulting room perspective. PMID- 17132348 TI - Primary care funding, contract status, and outcomes: an observational study. AB - BACKGROUND: The introduction of the Quality and Outcomes Framework (QOF) provides a quantitative way of assessing quality of care in general practice. We explore the achievements of general practice in the first year of the QOF, with specific reference to practice funding and contract status. AIM: To determine the extent to which differences in funding and contract status affect quality in primary care. DESIGN OF STUDY: Cross-sectional observational study using practice data obtained under the Freedom of Information Act 2000. SETTING: One hundred and sixty-four practices from six primary care trusts (PCTs) in England. METHOD: Practice data for all 164 practices were collated for income and contract status. The outcome measure was QOF score for the year 2004-2005. All data were analysed statistically. RESULTS: Contract status has an impact on practice funding, with Employed Medical Services (EMS) and Personal Medical Services (PMS) practices receiving higher levels of funding than General Medical Services (GMS) practices (P<0.001). QOF scores also vary according to contract status. Higher funding levels in EMS practices are associated with lower QOF scores (P=0.04); while GMS practices exhibited the opposite trend, with higher-funded practices achieving better quality scores (P<0.001). CONCLUSION: GMS practices are the most efficient contract status, achieving high quality scores for an average of pound 62.51 per patient per year. By contrast, EMS practices are underperforming, achieving low quality scores for an average of pound 105.37 per patient per year. Funding and contract status are therefore important factors in determining achievement in the QOF. PMID- 17132349 TI - Practice size and quality attainment under the new GMS contract: a cross sectional analysis. AB - BACKGROUND: The Quality and Outcomes Framework (QOF) of the new General Medical Services contract, for the first time, incentivises certain areas of general practice workload over others. The ability of practices to deliver high quality care may be related to the size of the practice itself. AIM: To explore the relationship between practice size and points attained in the QOF. DESIGN OF STUDY: Cross-sectional analyses of routinely available data. SETTING: Urban general practice in mainland Scotland. METHOD: QOF points and disease prevalence were obtained for all urban general practices in Scotland (n = 638) and linked to data on the practice, GP and patient population. The relationship between QOF point attainment, disease prevalence and practice size was examined using univariate statistical analyses. RESULTS: Smaller practices were more likely to be located in areas of socioeconomic deprivation; had patients with poorer health; and were less likely to participate in voluntary practice-based quality schemes. Overall, smaller practices received fewer QOF points compared to larger practices (P = 0.003), due to lower point attainment in the organisational domain (P = 0.002). There were no differences across practice size in the other domains of the QOF, including clinical care. Smaller practices reported higher levels of chronic obstructive pulmonary disease (COPD) and mental health conditions and lower levels of asthma, epilepsy and hypothyroidism. There was no difference in the reported prevalence of hypertension or coronary heart disease (CHD) across practices, in contrast to CHD mortality for patients aged under 70 years, where the mortality rate was 40% greater for single-handed practices compared with large practices. CONCLUSIONS: Although smaller practices obtained fewer points than larger practices under the QOF, this was due to lower scores in the organisational domain of the contract rather than to lower scores for clinical care. Single-handed practices, in common with larger practices serving more deprived populations, reported lower than expected CHD prevalence in their practice populations. Our results suggest that smaller practices continue to provide clinical care of comparable quality to larger practices but that they may need increased resources or support, particularly in the organisational domain, to address unmet need or more demanding QOF criteria. PMID- 17132350 TI - Workload and reward in the Quality and Outcomes Framework of the 2004 general practice contract. AB - BACKGROUND: The Quality and Outcomes Framework (QOF) of the 2004 UK General Medical Services (GMS) contract links up to 20% of practice income to performance measured against 146 quality indicators. AIM: To examine the distribution of workload and payment in the clinical domains of the QOF, and to compare payment based on true prevalence to the implemented system applying an adjusted prevalence factor. We aimed also to assess the performance of the implemented payment system against its three stated objectives: to reduce variation in payment compared to a system based on true prevalence, to fairly link reward to workload, and finally, to help tackle health inequalities. DESIGN OF STUDY: Retrospective analysis of publicly available QOF data. SETTING: Nine hundred and three GMS general practices in Scotland. METHOD: Comparison of payment under the implemented Adjusted Disease Prevalence Factor, and under an alternative True Disease Prevalence Factor. RESULTS: Variation in total clinical QOF payment per 1000 patients registered is significantly reduced compared to a payment system based on true prevalence. Payment is poorly related to workload in terms of the number of patients on the disease register, with up to 44 fold variation in payment per patient on the disease register for practices delivering the same quality of care. Practices serving deprived populations are systematically penalized under the implemented payment system, compared to one based on true prevalence. CONCLUSIONS: The implemented adjustment for prevalence succeeds in its aim of reducing variation in practice income, but at the cost of making the relationship between workload and reward highly inequitable and perpetuating the inverse care law. PMID- 17132351 TI - Impact on hypertension control of a patient-held guideline: a randomised controlled trial. AB - BACKGROUND: Hypertension is generally poorly controlled in primary care. One possible intervention for improving control is the harnessing of patient expertise through education and encouragement to challenge their care. AIM: To determine whether encouraging patients to manage their hypertension in an 'expert' manner, by providing them with information in a clear clinical guideline, coupled with an explicit exhortation to become involved in and to challenge their own care if appropriate, would improve their care. DESIGN OF STUDY: Single blind randomised controlled trial of detailed guideline versus standard information. SETTING: Single urban general practice over 1 year. METHOD: Patient-held guideline with written explicit exhortation to challenge care when appropriate. Two hundred and ninety-four of 536 eligible patients on the practice hypertension register were recruited, all of whom were randomised into one of two groups. Two hundred and thirty-six patients completed the study. RESULTS: PRIMARY OUTCOME: average systolic blood pressure. SECONDARY OUTCOMES: proportion of patients with blood pressure < 150 mmHg systolic and < 90 mmHg diastolic, average cholesterol, proportion of patients prescribed statins and aspirin according to guideline, hospital anxiety and depression score. No clinically, or statistically significant differences were found between intervention and control with respect to all parameters or in anxiety and depression levels. Statin and aspirin use improved throughout the course of the study in both groups. Statin use showed a trend (P = 0.02) in favour of control. CONCLUSION: In this study there was no clinically significant perceived benefit to patients as a result of providing them with a hypertension guideline. Patient guidelines are currently planned for many chronic illnesses. It is important to determine the utility of such interventions before scarce resources are applied to them. PMID- 17132352 TI - Hepatitis C infection among injecting drug users in general practice: a cluster randomised controlled trial of clinical guidelines' implementation. AB - BACKGROUND: Hepatitis C is a common infection among injecting drug users and has important implications for general practice. Although several clinical guidelines concerning the infection have been published, their effectiveness has yet to be tested. AIM: To assess the effectiveness of a general practice-based complex intervention to support the implementation of clinical guidelines for hepatitis C management among current or former drug users attending general practice. DESIGN OF STUDY: Cluster randomised controlled trial. SETTING: General practices in the Eastern Regional Health Authority area of Ireland. METHOD: Twenty-six practices were randomly allocated within strata to receive the intervention under study or to provide care as usual for a period of 6 months. There was screening for patients attending general practice for methadone maintenance treatment for hepatitis C and referral of anti-HCV antibody positive patients to a specialist hepatology department for assessment. RESULTS: At study completion, patients in the intervention group were significantly more likely to have been screened for hepatitis C than those in the control group, odds ratio adjusted for clustering 3.76 (95% confidence interval [CI] = 1.3 to 11.3) and this association remained significant after adjusting for other potentially confounding variables, using multiple logistic regression, with the odds ratio adjusted for clustering 4.53 (95% CI = 1.39 to 14.78). Although anti-HCV antibody positive patients in the intervention group were more likely to have been referred to a hepatology clinic, this was not statistically significant (P = 0.06). CONCLUSION: General practice has an important role in the care of people at risk of hepatitis C and when appropriately supported can effectively implement current best practice. PMID- 17132353 TI - Involve the patient and pass the MRCGP: investigating shared decision making in a consulting skills examination using a validated instrument. AB - BACKGROUND: Shared decision making is an important aspect of patient centredness. Lack of this consulting behaviour is a common reason for failure in the Membership of the Royal College of General Practitioners (MRCGP) consulting skills examination. AIM: To investigate candidates' performance in shared decision making and overall performance in the MRCGP consulting skills assessment compared with an independently validated measure, the OPTION ('observing patient involvement') scale. DESIGN: Cross-sectional study. SETTING: MRCGP examination, UK. PARTICIPANTS: Two hundred and fifty-two consultations submitted by 36 GPs submitting seven consultations per videotape. METHOD: A stratified sample of 63 candidates, 21 each from fail, pass and merit selected from candidates in the MRCGP consulting skills examination, were approached for participation. Participants' examination videotapes were independently assessed for shared decision making using the OPTION scale by two non-clinical raters. RESULTS: Thirty-six candidates (of 63; 57%) who participated were no different from non participants. Candidates who passed the 'sharing management options' in the MRCGP had significantly higher OPTION scores than those who did not (35.4 versus 27.3; mean difference = 8.1, P = 0.044). There was a significant difference between OPTION scores of MRCGP candidates with 'fail' and 'pass' (including pass with merit): 28.6 versus 36.1, 95% confidence interval CI = 1.13 to 13.87. Scores decreased as clinician age increased but were not significantly associated with sex of GP, age or sex of patient or consultation duration. The probability of passing the MRGCP increased as OPTION scores increased. CONCLUSION: This study demonstrated concurrent validity of the MRCGP consulting skills assessment of sharing management options against an independent validated instrument for shared decision making, the OPTION scale. Candidates who performed best in the MRCGP exhibited high scores with OPTION. This study provides the basis for further work to demonstrate evidence for the potential of training for professional assessment to improve consulting competence. PMID- 17132354 TI - Delay in diagnosis of lung cancer in general practice. AB - BACKGROUND: Lung cancer is a common cancer disease; nevertheless, in Denmark a GP only sees one new case of lung cancer per year. The core symptoms of lung cancer, cough and dyspnoea, are on the other hand very common in general practice. This represents a challenge to the diagnostic process and increases the risk of diagnostic delay. AIM: To explore diagnostic delay in primary health care among patients with lung cancer. DESIGN OF STUDY: A population based observational case series of 84 lung cancer patients' delay. SETTING: The County of Aarhus, Denmark. METHOD: From county-based registers of all histological and cytological tests we identified all patients in the County of Aarhus with lung cancer diagnosed during a 6 month period in 2003. Inclusion was verified by contacting the departments involved in the treatment of lung cancer. Data were based on telephone interviews with patients' GPs. A review of delay was made starting with the first symptom until referral to secondary care. RESULTS: The overall median delay in primary health care was 32.5 days (interquartile interval (IQI) = 12-68 days). One-third of the patients were referred to the diagnostic investigation after their first consultation. Important reasons for delay in primary care were: symptoms not related to the lungs, chest X-ray without suspicion of cancer, comorbidity, waiting times for investigations and lack of explicit follow-up appointment. Prolonged delay was often due to a combination of these factors. CONCLUSION: When diagnosing lung cancer, it is important that GPs are aware of the low sensitivity of chest X-ray. Shorter waiting times for investigations could reduce delay considerably. PMID- 17132355 TI - Communication between South Asian patients and GPs: comparative study using the Roter Interactional Analysis System. AB - BACKGROUND: The UK South Asian population has poorer health outcomes. Little is known about their process of care in general practice, or in particular the process of communication with GPs. AIM: To compare the ways in which white and South Asian patients communicate with white GPs. DESIGN OF STUDY: Observational study of video-recorded consultations using the Roter Interactional Analysis System (RIAS). SETTING: West Yorkshire, UK. METHOD: One hundred and eighty-three consultations with 11 GPs in West Yorkshire, UK were video-recorded and analysed. RESULTS: Main outcome measures were consultation length, verbal domination, 16 individual abridged RIAS categories, and three composite RIAS categories; with comparisons between white patients, South Asian patients fluent in English and South Asian patients nonfluent in English. South Asians fluent in English had the shortest consultations and South Asians non-fluent in English the longest consultations (one-way ANOVA F = 7.173, P = 0.001). There were no significant differences in verbal domination scores between the three groups. White patients had more affective (emotional) consultations than South Asian patients, and played a more active role in their consultations, as did their GPs. GPs spent less time giving information to South Asian patients who were not fluent in English and more time asking questions. GPs spent less time giving information to South Asian patients fluent in English compared with white patients. CONCLUSIONS: These findings were expected between patients fluent and non-fluent in English but do demonstrate their nature. The differences between white patients and South Asian patients fluent in English warrant further explanation. How much of this was due to systematic differences in behaviour by the GPs, or was in response to patients' differing needs and expectations is unknown. These differences may contribute to differences in health outcomes. PMID- 17132356 TI - The effect of interventions to alter the consultation length of family physicians: a systematic review. AB - BACKGROUND: Observational studies have shown differences in process and outcome between the consultations of primary care physicians whose average length of consultation differs, however, these differences may be due to self selection. AIM: To assess the effectiveness and cost effectiveness of interventions to alter primary care physicians' consultation length. DESIGN OF STUDY: Systematic review with narrative analysis. METHOD: Data sources included Medline, EMBASE, the Cochrane Controlled Trials Register and Effective Practice and Organisation of Care Group specialised register, the NHS National Research Register and author contacts. To be eligible, studies had to be controlled trials. They had to evaluate interventions to alter the consultation length of primary care physicians, and provide objectively measured process or outcome data. Data were extracted independently using agreed criteria and disagreements resolved by discussion. RESULTS: Six articles describing four trials were included. All took place in the UK and tested short term changes in the time allocated to each patient, and all had methodological weaknesses, particularly due to non random allocation of patients. Altering appointment length resulted in modest changes in average consultation length. There were no consistent differences in problem recognition, examination, prescribing, referral or investigation rates. There was some evidence that blood pressure was checked more frequently and smoking discussed more often when more time was available. None of the interventions were associated with differences in patient satisfaction. No trials examined cost effectiveness. CONCLUSIONS: Our findings do not provide sufficient evidence to support or resist a policy of altering consultation lengths of primary care physicians. Further trials are needed, focussing on health outcomes and cost effectiveness. PMID- 17132357 TI - The use of cardiovascular risk factor information in practice databases. PMID- 17132358 TI - Letter from a Dutch uncle. PMID- 17132360 TI - Privatising primary care. PMID- 17132359 TI - Domestic violence in practice. PMID- 17132361 TI - Ever been HAD? PMID- 17132362 TI - Chronic kidney disease. PMID- 17132363 TI - Warfarin for atrial fibrillation. PMID- 17132364 TI - Malignancy and deep vein thrombosis. PMID- 17132366 TI - Physical activity may be good for you but we are not the key players. PMID- 17132368 TI - Setting up a primary care STI detection and treatment service in Dili, East Timor. PMID- 17132369 TI - With appropriate incentives, general practice can improve the coverage of the National Chlamydia Screening Programme. PMID- 17132370 TI - How to die--a physician's last lesson. PMID- 17132371 TI - Foundation Year 2: changing attitudes towards general practice. PMID- 17132376 TI - Self-care--has DIY gone too far? PMID- 17132377 TI - Chronic kidney disease: a new priority for primary care. PMID- 17132378 TI - Preferences for self-care or professional advice for minor illness: a discrete choice experiment. AB - AIM: To determine the relative importance of factors that influence decision making in the management of minor illness, and how people trade between these factors. DESIGN OF STUDY: Discrete choice experiment. SETTING: Scottish electoral roll. METHOD: Six hundred and fifty-two responders of a previous national survey were invited to complete a discrete choice experiment questionnaire. This was used to measure relative preferences for managing symptoms of minor illness often associated with analgesic use. Three attributes were identified as important to participants: type of management, availability, and cost of managing symptoms. Trade-offs between these attributes were examined. RESULTS: A 57% response rate was achieved (51% valid response rate). People preferred to manage symptoms by self-care and were willing to pay almost pounds 23 to do so. Community pharmacy was the preferred source of advice. Responders preferred less waiting time and paying less money when managing symptoms, and were willing to trade between factors. A less preferred type of management became more attractive when waiting times and cost were reduced. CONCLUSION: Findings suggest that self-care is the preferred method of managing symptoms of minor illness. When developing services to support self-care, policy makers should invest in services that reduce waiting times and incur least cost to users. PMID- 17132379 TI - Implementation of Advanced Access in general practice: postal survey of practices. AB - BACKGROUND: Advanced Access has been strongly promoted as a means of improving access to general practice. Key principles include measuring demand, matching capacity to demand, managing demand in different ways and having contingency plans. Although not advocated by Advanced Access, some practices have also restricted availability of pre-booked appointments. AIM: This study compares the strategies used to improve access by practices which do or do not operate Advanced Access. DESIGN OF STUDY: Postal survey of practices. SETTING: Three hundred and ninety-one practices in 12 primary care trusts. METHOD: Questionnaires were posted to practice managers to collect data on practice characteristics, supply and demand of appointments, strategies employed to manage demand, and use of Advanced Access. RESULTS: Two hundred and forty-five from 391 (63%) practices returned a questionnaire and 162/241(67%) claimed to be using Advanced Access. There were few differences between characteristics of practices operating Advanced Access or not. Both types of practice had introduced a wide range of measures to improve access. The proportion of doctors' appointments only available for booking on the same day was higher in Advanced Access practices (40 versus 16%, difference = 24%, 95% CI = 16% to 32%). Less than half the practices claiming to operate Advanced Access ((63/140; 45%) used all four of this model's key principles. CONCLUSION: The majority of practices in this sample claim to have introduced Advanced Access, but the degree of implementation is very variable. Advanced Access practices use more initiatives to measure and improve access than non-Advanced Access practices. PMID- 17132380 TI - Young adults' perceptions of GPs as a help source for mental distress: a qualitative study. AB - BACKGROUND: Few young adults with mental disorder seek help from a GP. AIM: To explore young adults' perceptions of GPs as a source of help for mental distress. DESIGN OF STUDY: Qualitative interviews. SETTING: Bristol and surrounding areas, UK. METHOD: Males and females aged 16-24 years screened as 'cases' with probable mental disorder (GHQ [General Health Questionnaire]-12 score> or =4) or describing past episodes of mental disorder (n = 23) were sampled purposively according to help-seeking behaviour. Semi-structured interviews explored help seeking choices. Transcripts were analysed using thematic, constant comparison and case study analysis. RESULTS: Most young adults did not value or recognise GPs as a source of help for mental disorder or distress. They thought that GPs deal exclusively with physical illness, lack training in mental health, are unable to provide 'talking' therapy, and may be dismissive of those consulting with mental distress. A prescription for antidepressants was seen as the most likely outcome of a consultation, but young adults wished to avoid this and so rarely consulted. Encounters with GPs could challenge or reinforce these perceptions. CONCLUSION: Negative perceptions about the value of consulting a GP for mental distress may explain low rates of help-seeking among young adults, including those with severe distress. Young people require a better understanding of GPs' role. It is also necessary to address evidence reported elsewhere that some GPs also experience uncertainties about what they can offer within the constraints of primary care. PMID- 17132381 TI - Symptomatic peripheral arterial disease: the value of a validated questionnaire and a clinical decision rule. AB - BACKGROUND: If a validated questionnaire, when applied to patients reporting with symptoms of intermittent claudication, could adequately discriminate between those with and without peripheral arterial disease, GPs could avoid the diagnostic measurement of the ankle brachial index. AIM: To investigate the Edinburgh Claudication Questionnaire (ECQ) in general practice and to develop a clinical decision rule based on risk factors to enable GPs to easily assess the likelihood of peripheral arterial disease. DESIGN OF STUDY: An observational study. SETTING: General practice in The Netherlands. METHOD: This observational study included patients of > or =55 years visiting their GP for symptoms suggestive of intermittent claudication or with one risk factor. The ECQ and the ankle brachial index were performed. The prevalence of peripheral arterial disease, defined as an ankle brachial index <0.9, was related to risk factors using logistic regression analyses, on which a clinical decision rule was developed and related to the presence of peripheral arterial disease. RESULTS: Of the 4790 included patients visiting their GP with symptoms suggestive of intermittent claudication, 4527 were eligible for analyses. The prevalence of peripheral arterial disease in this group was 48.3%. The sensitivity of the ECQ was only 56.2%. The prevalence of peripheral arterial disease in a clinical decision rule that included age, male sex, smoking, hypertension, hypercholesterolemia, and a positive ECQ, increased from 14% in the lowest to 76% in the highest category. CONCLUSION: This study indicates that the ECQ alone has an inadequate diagnostic value in detecting patients with peripheral arterial disease. The ankle brachial index should be performed to diagnose peripheral arterial disease in patients with complaints suggestive of intermittent claudication, although our clinical decision rule could help to differentiate between extremely high and lower prevalence of peripheral arterial disease. PMID- 17132382 TI - The effectiveness of a treatment protocol for male lower urinary tract symptoms in general practice: a practical randomised controlled trial. AB - BACKGROUND: Randomised controlled trials have shown the efficacy of several treatment modalities for lower urinary tract symptoms (LUTS) in selected populations. The effectiveness in daily practice has hardly been investigated, especially in primary care and is dependent on choices between all possible treatment options and best investigated in a comprehensive study, including all treatment modalities (watchful waiting, alpha-blockers, 5-alpha-reductase inhibitors, and surgery). AIM: Assessment of the effectiveness of a comprehensive treatment protocol for LUTS in primary care. DESIGN OF STUDY: Randomised controlled trial. SETTING: Fourteen general practices in the Netherlands. METHOD: INTERVENTION: treatment protocol based on a formalised expert opinion. Control condition: usual care. STUDY POPULATION: 208 subjects with moderate to severe LUTS (IPSS > or =8, median = 13). OUTCOME MEASURES: symptom severity (IPSS [International Prostate Symptom Score]), bother score (Dan-PSS [Danish Prostate Symptom Score]), and maximum urinary flow (Q(max)); incidence of acute urinary retention and urinary tract infections. RESULTS: In the intervention group markedly more subjects used an alpha-blocker at end of follow-up than in the usual care group (24% versus 6%). No significant differences were found between intervention and control group in IPSS, Q(max) or Dan-PSS. CONCLUSION: alpha blockers and watchful waiting are the most frequent treatment modalities for LUTS in primary care. Our study showed no evidence that a protocol using well-defined indications for all possible treatment modalities based on a formalised expert opinion procedure has added value. Based on our results, we cannot recommend a broadening of the indication for alpha-blockers, which, however, seems to be the current trend. PMID- 17132383 TI - Urinary incontinence in Moroccan and Turkish women: a qualitative study on impact and preferences for treatment. AB - AIM: To gain insight into the impact of urinary incontinence on the daily lives of Moroccan and Turkish women and their preferences for treatment. DESIGN OF STUDY: A qualitative analysis of data from semi-structured in depth interviews with 30 Moroccan and Turkish migrant women with urinary incontinence. SETTING: Six general practices with large numbers of immigrant families in four cities in the Netherlands. METHOD: Semi-structured in depth interviews were conducted with Moroccan and Turkish migrant women, who presented with complaints of urinary incontinence in six general practices with large numbers of immigrant families on the practice list in four different cities in the Netherlands. RESULTS: All the women adhered closely to bodily cleanliness and considered incontinence to be dirty. As Muslims, they were obliged to perform ritual prayers preceded by ablution five times per day and the urinary incontinence breached their status of ritual purity. Therefore, they had to wash more often and experienced this as a heavy burden. In a number of the women, shame formed a reason why they could not talk to anybody about the incontinence, not even with the doctor. One-third of the women felt that their GP had not taken them seriously. Knowledge about anatomy, physiology and available treatments was mostly lacking. In addition, the women did not understand the aim of the exercises from the physiotherapist. The majority of women gave preference to help from a female doctor. CONCLUSION: Urinary incontinence in Moroccan and Turkish migrant women formed a considerable problem in their daily following of the Islam faith. Shame on the part of the patient and miscommunication at the doctor's surgery led to inadequate care. PMID- 17132384 TI - Menstrual symptoms: the importance of social factors in women's experiences. AB - BACKGROUND: Menstrual disorders are a common presentation in primary care. Heavy menstrual bleeding is the most common concern, and is often treated by medical and surgical means despite lack of pathology. AIM: To explore women's experiences of menstrual disorders. DESIGN OF STUDY: Two qualitative studies using semi structured interviews. SETTING: Inner-city London. METHOD: An initial study recruited women with heavy menstrual bleeding via their GPs. A follow-up study recruited women with a variety of menstrual problems via general practice and the community. Interviews were taped and transcribed then analysed using the constant comparative method. RESULTS: Management of menstruation was a prominent theme in interviews. Women acted to comply with a strong social message that menstruation should be concealed, although this behaviour was often 'taken for granted.' The need to conceal evidence or reminders of menstrual bleeding was particularly important. Onset of menstrual symptoms often challenged established strategies for menstrual management. Menstrual management then became a conscious problem and a source of continuing stress. The breakdown of management strategies, by real or threatened episodes of leaking or staining, influenced consultation behaviour and decisions about treatment. CONCLUSION: The social pressure to maintain concealment of menstruation is a strong influence on women's health related behaviour in response to menstrual concerns. Women's choices may be better understood if attention is paid to the social context in which they live. PMID- 17132385 TI - Withdrawal from long-term benzodiazepine use: randomised trial in family practice. AB - BACKGROUND: The long-term use of benzodiazepines is highly prevalent in developed societies and is not devoid of risks. Withdrawing patients from these drugs is often difficult. Tapering off benzodiazepines has been shown to be a good strategy for discontinuing their long-term use. AIM: To establish the efficacy of an intervention programme for reducing the chronic use of benzodiazepines. DESIGN OF STUDY: Randomised, two-arm, parallel, non-blinded controlled trial. SETTING: Three urban healthcare centres covering a population of 50,000 inhabitants (Mallorca, Spain). METHOD: Adult patients (n = 139) taking benzodiazepines daily for more than a year and visited by their family physician were randomised into an intervention group (n = 73) that received standardised advice and a tapering off schedule with biweekly follow-up visits, or into a control group (n = 66), that was managed following routine clinical practice. Both were followed for a year. RESULTS: Patients achieved withdrawal or reduced their dose by at least 50% after 6 and 12 months. Abstinence and withdrawal symptoms were also measured. Both groups were homogeneous for personal, clinical and psychological characteristics and for benzodiazepine use. Only two patients from each group were lost to follow-up. After 12 months, 33 (45.2%) patients in the intervention group and six (9.1%) in the control group had discontinued benzodiazepine use; relative risk = 4.97 (95% confidence interval [CI] = 2.2 to 11.1), absolute risk reduction = 0.36 (95% CI = 0.22 to 0.50). For every three interventions, one patient achieved withdrawal. Sixteen (21.9%) subjects from the intervention group and 11 (16.7%) controls reduced their initial dose by more than 50%. CONCLUSION: Standardised advice given by the family physician, together with a tapering off schedule, is effective for withdrawing patients from long-term benzodiazepine use and is feasible in primary care. PMID- 17132386 TI - GPs' attitudes to benzodiazepine and 'Z-drug' prescribing: a barrier to implementation of evidence and guidance on hypnotics. AB - Zaleplon, zolpidem, and zopiclone ('Z-drugs') prescribing is gradually rising in the UK, while that of benzodiazepine hypnotics is falling. This situation is contrary to current evidence and guidance on hypnotic prescribing. The aim of this study was to determine and compare primary care physicians' perceptions of benefits and risks of benzodiazepine and Z-drug use, and physicians' prescribing behaviour in relation to hypnotics using a cross-sectional survey. In 2005 a self administered postal questionnaire was sent to all GPs in West Lincolnshire Primary Care Trust. The questionnaire investigated perceptions of benefits and disadvantages of benzodiazepines and Z-drugs. Of the 107 questionnaires sent to GPs, 84 (78.5%) analysable responses were received. Responders believed that Z drugs were more effective than benzodiazepines in terms of patients feeling rested on waking (P<0.001), daytime functioning (P<0.001), and total sleep time (P = 0.03). Z-drugs were also thought to be safer in terms of tolerance (P<0.001), addiction (P<0.001), dependence (P<0.001), daytime sleepiness (P<0.001), and road traffic accidents (P = 0.018), and were thought to be safer for older people (P<0.001). There were significant differences between GPs' perceptions of the relative benefits and risk of Z-drugs compared with benzodiazepines. The majority of practitioners attributed greater efficacy and lower side effects to Z-drugs. GPs' beliefs about effectiveness and safety are not determined by current evidence or national (NICE) guidance which may explain the increase in Z-drug prescribing relative to benzodiazepine prescribing. PMID- 17132387 TI - Using common ophthalmologic jargon in correspondence can lead to miscommunication. PMID- 17132388 TI - Appropriate postgraduate training. PMID- 17132389 TI - Early diagnosis of COPD. PMID- 17132390 TI - Questions about COPD. PMID- 17132391 TI - The support of obese patients in primary care. PMID- 17132392 TI - The good lie? PMID- 17132393 TI - Childhood obesity. PMID- 17132394 TI - 15,000 times 15 minutes. PMID- 17132395 TI - Not tonight Josephine: on private incursions into primary care. PMID- 17132396 TI - The 'Mensch' factor in general practice: a role to demonstrate professionalism to students. PMID- 17132398 TI - NICE as oxymoron. PMID- 17132397 TI - Nature therapy. PMID- 17132403 TI - The Future of the International Federation for the Surgery of Obesity (IFSO). PMID- 17132404 TI - Nonalcoholic fatty liver disease of obesity. AB - Nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) are part of the same continuum. They are a major, under-recognized cause of chronic liver disease. Good medical treatment options do not exist to date. The mainstay of treatment is weight loss. Bariatric surgery offers weight loss and improvement of NAFLD and NASH. PMID- 17132405 TI - Revision of failed vertical banded gastroplasty to non-resectional Scopinaro biliopancreatic diversion: early experience. AB - BACKGROUND: The most commonly performed revision operation following failed vertical banded gastroplasty (VBG) is Roux-en-Y gastric bypass, although revision to biliopancreatic diversion (BPD) with duodenal switch is now another common option. We describe the surgical technique for revision of a failed VBG to a non resectional Scopinaro BPD in a series of patients, as well as the outcome in terms of complications and mean % excess weight loss (%EWL). METHODS: A retrospective review was conducted on all patients who underwent revision to BPD at Mercy Bariatrics, Western Australia, between June 2001 and April 2005. This yielded 20 patients who had revision to BPD, 9 of whom had VBG as their initial operation. The mean %EWL was measured at regular intervals postoperatively (3, 6, 12, and 24 months). RESULTS: Mean %EWL at 12 and 24 months was 69.5 and 76.7, respectively. These results are comparable to %EWL after a primary BPD. Nutritional manifestations were found to be the most common of the minor complications. CONCLUSION: Our technique for revision of a failed restrictive operation to a non-resectional Scopinaro BPD is described. The preliminary results in terms of %EWL and complications are comparable to other revisional malabsorptive operations. Prospective randomized controlled trials are needed to further evaluate effects of revision to a non-resectional Scopinaro BPD and to ensure that the results (in terms of %EWL) are reproducible. PMID- 17132406 TI - Serum ghrelin, leptin and adiponectin levels before and after weight loss: comparison of three methods of treatment--a prospective study. AB - BACKGROUND: Ghrelin is a peptide hormone with orexigenic properties, primarily produced by the stomach. Leptin and adiponectin are the two adiposity products that participate in body weight control. Leptin always decreases and adiponectin increases after weight loss. Different changes in fasting ghrelin levels have been reported following bariatric surgery. In this study, we compare the changes in fasting ghrelin, leptin and adiponectin levels in 3 groups of patients who achieved weight loss by either diet, MacLean vertical banded gastroplasty (VBG) or biliopancreatic diversion with duodenal switch (BPD-DS). METHODS: Serum fasting ghrelin, leptin and adiponectin concentration was measured in 40 obese patients who achieved weight loss by either diet (n=14), VBG (n=13) or BPD-DS (n=13), before and after weight loss. The follow-up period was 18 months for BPD DS and VBG and 6 months for diet. Serum ghrelin level was measured by ELISA. RESULTS: BMI was significantly decreased in all 3 groups: 9.2+/-2.4% (P<0.01) following diet, 38.47+/-7.26% (P<0.01) after VBG, and 42.88+/-9.09% after BPD-DS (P<0.01). Serum fasting ghrelin level increased after diet (110.45+/-117.84%, P=0.002) and VBG (65.48+/-92.93%, P=0.001),but decreased after BPD-DS (-21.63+/ 28.63%, P=0.019). Leptin concentration decreased and adiponectin increased in all groups. CONCLUSIONS: Unlike after diet or gastric restrictive surgery, BPD-DS is associated with markedly suppressed ghrelin levels, possibly contributing to the weight-reducing effect of this operation. Sleeve gastrectomy seems to be the main cause of this reduction. PMID- 17132407 TI - Effectiveness of biliopancreatic diversion in the patients with bulimia nervosa. AB - BACKGROUND: Bulimia nervosa (BN) is an eating disorder, characterized by consumption of huge amounts of food during discrete periods. Unlike patients with binge-eating disorder (BED), patients with BN demonstrate elements of compensatory "purging" behavior to prevent weight gain and obesity: i.e. self induced vomiting, use of laxatives and enemas. These habits may prevent patients from attaining morbid obesity (MO), but may seriously affect life-style and become an excruciating, sometimes life-threatening condition. METHODS: 6 of 108 patients (5.6%) who underwent BPD in our clinic (laparoscopic Scopinaro BPD - 1, open BPD/DS - 4, Lap. BPD/DS - 1) suffered from BN preoperatively. Their preoperative weight was 68-117 kg and BMI 27.6-41.9 kg/m(2). 4 of 6 patients had BMI <40 kg/m(2) before BPD but were MO in the past. The patient with the lowest weight had repeated gastroesophageal bleeding during self-induced vomiting. 3 of the 6 patients had previously failed intragastric balloon or Lap-Band. RESULTS: All 6 patients were cured or significantly improved of bulimic symptoms soon after BPD. Weight loss was very good and never reached an undesirably low level. Patient satisfaction was high. CONCLUSION: Severe BN may be considered as a latent and potentially malignant MO. BPD (or BPD/DS) may be an effective solution for some patients with severe BN, as a final decision after unsuccessful organized conservative attempts. All candidates for bariatric surgery should be screened for BN, because it may influence choice of procedure in favor to BPD or BPD/DS. PMID- 17132408 TI - Type 2 diabetes and weight loss following biliopancreatic diversion for obesity. AB - BACKGROUND: The authors investigated the weight loss and maintenance in type 2 diabetic obese patients undergoing biliopancreatic diversion (BPD). METHODS: Two series of diabetic and non-diabetic obese patients matched for gender, age and baseline body mass index (BMI) were evaluated prior to BPD, on the occasion of the regular follow-up visit at 1, 2 and 3 years following the operation, and at the fifth postoperative year. At each follow-up point, body weight (BW), BMI, and serum glucose concentration were measured. RESULTS: In all type 2 diabetic patients, the serum glucose level fell to within the normal range at the first postoperative year and remained within normal limits without any medication throughout all the follow-up period. In preoperatively diabetic subjects, mean values of BW and BMI were closely similar to those of non-diabetic subjects at all follow-up points, and the stabilization weight was independently related to age and to initial BW values. CONCLUSIONS: In obese patients with type 2 diabetes, the glucose level steadily normalized in every case following BPD, and values remained unchanged throughout the follow-up period. After the operation, the type 2 diabetic obese patients experienced the same stable weight reduction as their non-diabetic counterparts. PMID- 17132409 TI - Sleeve gastrectomy in the high-risk patient. AB - BACKGROUND: One of the surgical options available for the super-obese patient is the sleeve gastrectomy. We present results of this operation in a series of 118 patients. METHODS: The charts of all patients who have had the sleeve gastrectomy performed were reviewed for demographic data, complications, weight, and nutritional parameters. RESULTS: Median age was 47 years (16-70). Median BMI was 55 kg/m(2) (37-108), with 73% of patients having a BMI > or =50 kg/m(2). 41% of the patients were male. The operation was performed by laparotomy in all but three cases, which were performed laparoscopically. Median hospital stay was 6 days (3-59). There was one perioperative death (0.85%). 18 patients (15.3%) had postoperative complications. Median percent excess weight loss was 37.8% at 6 months, 49.4% at 12 months, and 47.3% at 24 months. Median follow-up was 13 months (1-66). At 1 year postoperatively, the percentage of patients with normal serum levels of albumin was 100%, hemoglobin 86.1%, and calcium 87.2%, compared to 98.1%, 85.6%, and 94.3% preoperatively. 6 patients requested conversion to a duodenal switch during the follow-up period; all left the hospital in 4-6 days without major complication. CONCLUSIONS: Although the sleeve gastrectomy does not result in as much weight loss as the duodenal switch or gastric bypass, it can be used as a stand-alone operation or as a bridge to more complex procedures in the high-risk super-obese patient. PMID- 17132410 TI - A prospective randomized study between laparoscopic gastric banding and laparoscopic isolated sleeve gastrectomy: results after 1 and 3 years. AB - BACKGROUND: Laparoscopic adjustable gastric banding (GB) is the most popular restrictive procedure for obesity in Europe. Isolated sleeve gastrectomy (SG), is less common, but more invasive and with a higher learning curve. The aim of this prospective randomized study was to compare the results of GB and SG after 1 and 3 years of surgery. METHODS: 80 patient candidates for laparoscopic restrictive surgery were operated consecutively and randomly, between January and December 31, 2002, by GB (7M, 33F) or by SG (9M, 31F) (NS). Median age was 36 (20-61) for GB versus 40 (22-65) for SG (NS). Median BMI was 37 (30-47) for GB versus 39 (30 53) for SG (NS). After 1 and 3 years: weight loss, feeling of hunger, sweet eating, gastroesophageal reflux disease (GERD), complications and re-operations were recorded in both groups. RESULTS: Median weight loss after 1 year was 14 kg (-5 to +38) for GB and 26 kg (0 to 46) for SG (P<0.0001); and after 3 years was 17 kg (0 to 40) for GB and 29.5 kg (1 to 48) for SG (P<0.0001). Median decrease in BMI after 1 year was 15.5 kg/m 2 (5 to 39) after GB and 25 kg/m(2) (0 to 45) after SG (P<0.0001); and after 3 years was 18 kg/m(2) (0 to 39) after GB and 27.5 kg/m 2 (0 to 48) after SG (P=0.0004). Median %EWL at 1 year was 41.4% (-11.8 to +130.5) after GB and 57.7% (0 to 125.5) after SG (P=0.0004); and at 3 years was 48% (0 to 124.8) after GB and 66% (-3.1 to +152.4) after SG (P=0.0025). Loss of feeling of hunger after 1 year was registered in 42.5% of patients with GB and in 75% of patients with SG (P=0.003); and after 3 years in 2.9% of patients with GB and 46.7% of patients with SG (P<0.0001). Loss of craving for sweets after 1 year was achieved in 35% of patients with GB and 50% of patients with SG (NS); and after 3 years in 2.9% of patients with GB and 23% of patients with SG (NS). GERD appeared de novo after 1 year in 8.8% of patients with GB and 21.8% of patients with SG (NS); and after 3 years in 20.5% of patients with GB and 3.1% of patients with SG (NS). Postoperative complications requiring re-operation were necessary for 2 patients after SG. Late complications requiring re-operation after GB included 3 pouch dilations treated by band removal in 2 and 1 laparoscopic conversion to Roux-en-Y gastric bypass (RYGBP), 1 gastric erosion treated by conversion to RYGBP, and 3 disconnections of the system treated by reconnection. Inefficacy affected 2 patients after GB, treated by conversion into RYGBP and 2 patients after SG treated by conversion to duodenal switch. CONCLUSION: Weight loss and loss of feeling of hunger after 1 year and 3 years are better after SG than GB. GERD is more frequent at 1 year after SG and at 3 years after GB. The number of re-operations is important in both groups, but the severity of complications appears higher in SG. PMID- 17132411 TI - The impact of bariatric surgery on menstrual patterns. AB - BACKGROUND: Obesity and anovulation are common medical problems in the United States. Anovulation in obese patients primarily manifests with irregular, sporadic or absent menstrual bleeding. Weight loss of at least 5% has been shown to reverse obesity-related anovulation. The aim of this study was to assess the impact of bariatric surgery on infertility in morbidly obese women and to identify factors associated with return of normal menses following bariatric surgery. METHODS: A survey of patients was collected from the bariatric surgery data-base at the Hospital of the University of Pennsylvania. 410 women under the age of 40 were sent questionnaires. 195 patients completed the questionnaire, and 29 patients had incorrect addresses without a forwarding address, resulting in a 51.2% response rate. Patients who reported menstrual cycle lengths >35 days were considered abnormal. 92 of the 195 responders were considered anovulatory preoperatively, based on menstrual history. RESULTS: There was no significant difference in postoperative BMI, BMI decrease or age at surgery between the survey responders and non-responders. There was a significant difference between these 2 groups in time since surgery (P=.01). Both groups had a decrease in BMI of >18 kg/m(2). The mean menstrual cycle length preoperatively among those categorized as ovulatory and anovulatory was 27.3 and 127.5 days, respectively. Of the 98 patients who were anovulatory preoperatively, 70 patients (71.4%) regained normal menstrual cycles after surgery. Those patients who regained ovulation had greater weight loss than those who remained anovulatory (61.4 kg vs 49.9 kg, P=0.02). CONCLUSIONS: Anovulation resulting in abnormal menses is a common problem in morbidly obese premenopausal women. The menstrual cycle disorders may completely resolve after bariatric surgery. Thus, infertility due to anovulation among morbidly obese women could potentially be viewed as an additional indication for bariatric surgery. PMID- 17132412 TI - Attitude of prospective surgical residents regarding surgery for morbid obesity. AB - BACKGROUND: Obesity and associated co-morbidities have become an epidemic in the United States. As surgery for obesity becomes more common, surgical training programs need to address this growing demand. We conducted this study to assess prospective surgery trainees' attitudes and knowledge regarding surgery for morbid obesity. METHODS: An anonymous and voluntary questionnaire was given to prospective surgical residency applicants to complete during their interview. The questionnaire included basic demographic information and addressed the applicants' attitudes and basic knowledge about surgery for obesity. RESULTS: 57 applicants to the surgical residency program completed the survey. Demographic information included: 51% male, 36% from the Northeast, 32% with obese family members, and 93% applying for a categorical surgery position. 81% of applicants had been exposed to bariatric surgery. Although 70% of applicants would perform bariatric surgery as part of their practice, only 44% would make this their career. Reasons for reluctance to treat bariatric surgery patients included: more complications (46%), non-compliant patients (33%), and technically demanding surgery (18%). 89% responded that they would recommend bariatric surgery to a family member, but only 77% would consider it for themselves. Overall correct answers regarding bariatric surgery knowledge were 74%. CONCLUSIONS: Attitudes and knowledge about surgery for morbid obesity were generally positive in prospective surgical trainees. Medical school curriculum and surgical training programs should continue to expose trainees to information from this ever-growing field. PMID- 17132413 TI - Does gastric banding for morbid obesity reduce or increase gastroesophageal reflux? AB - BACKGROUND: Conflicting results regarding the influence of laparoscopic adjustable gastric banding (LAGB) on gastroesophageal reflux disease (GERD) have been published. METHODS: A prospective follow-up study was conducted in 31 patients (male/female 5/26, mean age 44 +/- 11 SD years) with 24-hour pH and manometry recordings, symptom assessment, and upper GI endoscopy. RESULTS: Total number of reflux episodes decreased from a mean value of 44.6 +/- 23.7 SD preoperatively to 22.9 +/- 17.1 postoperatively (P=0.0006), after a median follow up time of 19 months (range 7-32 months). Total reflux time decreased from 9.5% +/- 6.2% to 3.5% +/- 3.7%, P=0.0009, and DeMeester score decreased from 38.5 +/- 24.9 to 18.6 +/- 20.4, P=0.03. Symptomatic patients decreased from 48.4% preoperatively to 16.1% postoperatively (P=0.01), medication for GERD decreased from 35.5% to 12.9% (P=0.05), and the diagnosis of GERD on 24-hour pH recordings decreased from 77.4% to 37.5% (P=0.01). There were no pouch enlargements seen on upper GI endoscopy. Esophageal motility was unchanged, but 36% of the patients had incomplete relaxation of the lower esophageal sphincter following the operation (P<0.0001). Mean BMI decreased from 46.0 +/- 5.46 to 38.4 +/- 6.45 (P<0.0001), excess weight from 60.0 kg +/- 18.58 kg, 44.9% +/- 6.56% to 38.4 kg +/- 20.27 kg, 28.4% +/- 10.97% (P<0.0001). No association between the postoperative diagnosis of GERD and the amount of weight loss could be found. CONCLUSIONS: The correctly placed gastric band is an effective anti-reflux barrier in the short term. Long-term results have to be awaited. PMID- 17132414 TI - Serum amyloid A and obstructive sleep apnea syndrome before and after surgically induced weight loss in morbidly obese subjects. AB - BACKGROUND: Serum amyloid A (SAA) is an inflammatory marker associated with cardiovascular disease (CVD) and found to be increased in obesity. Obstructive sleep apnea (OSA) syndrome, a frequent complication of obesity also associated with CVD risk, is improved after surgically-induced weight loss. To explore the potential role of SAA in the relation between OSA and CVD, we investigated relationships between changes in SAA concentrations and nocturnal respiratory events in obese subjects undergoing bariatric surgery. METHODS: We measured plasma SAA and used nocturnal respiratory polygraphy to assess the apneahypopnea index (AHI), the oxygen desaturation index (ODI) and the mean and lowest O(2) saturation (SaO(2) ) in 61 morbidly obese patients before either adjustable gastric banding or gastric bypass. For 35 subjects with OSA, the same data were obtained 1 year after the surgery. RESULTS: Before surgery, SAA concentrations were significantly higher in patients with severe OSA (56.2+/-6.4 microg/ml) compared to subjects with moderate OSA (22.9+/-3.2 microg/ml) or without OSA (16.2+/-2.2 microg/ml). Plasma SAA correlated positively with AHI and ODI, and negatively with mean and lowest SaO(2). After surgery, plasma SAA decreased significantly by 41.7%, and changes in plasma SAA correlated with variations in OSA parameters. In multivariate analyses, AHI was a predictor of plasma SAA, independent of BMI, both at baseline and during weight loss. CONCLUSION: The improvement of OSA after bariatric surgery is associated with a decrease in SAA, independent of the change in BMI. SAA may represent a marker of the improvement in CVD risk profile after surgically-induced weight loss in patients with OSA. PMID- 17132415 TI - Internal hernia after laparoscopic Roux-en-Y gastric bypass for morbid obesity: a continuous challenge in bariatric surgery. AB - BACKGROUND: Roux-en-Y gastric bypass (RYGBP) has long been associated with the possible development of internal hernias, with a reported incidence of 1-5%. Because it induces fewer adhesions than laparotomy, the laparoscopic approach to this operation appears to increase the rate of this complication, which can present dramatically. METHODS: Data from all patients undergoing bariatric surgery are introduced prospectively in a data-base. Patients who were reoperated for symptoms or signs suggestive of an internal hernia were reviewed retrospectively, with special emphasis on clinical and radiological findings, and surgical management. RESULTS: Of 607 patients who underwent laparoscopic primary or reoperative RYGBP in our two hospitals between June 1999 and January 2006, 25 developed symptoms suggestive of an internal hernia, 2 in the immediate postoperative period, and 23 later on, after a mean of 29 months and a mean loss of 14.5 BMI units. 9 of the latter presented with an acute bowel obstruction, of which 1 required small bowel resection for necrosis. Recurrent colicky abdominal pain was the leading symptom in the others. Reoperation confirmed the diagnosis of internal hernia in all but 1 patient. The most common location was the meso jejunal mesenteric window (16 patients, 56%), followed by Petersen's window (8 patients, 27%), and the mesocolic window (5 patients, 17%). Patients in whom the mesenteric windows had been closed using running non-absorbable sutures had fewer hernias than patients treated with absorbable sutures at the primary procedure (1.3% versus 5.6%, P=0.03). Except in the acute setting, clinical and radiological findings were of little help in the diagnosis. CONCLUSIONS: Except in the setting of acute obstruction, clinical and radiological findings usually do not help in the diagnosis of internal hernia. A high index of suspicion, based mainly on the clinical history of recurrent colicky abdominal pain, is the only means to reduce the number of acute complications leading to bowel resection by offering the patient an elective laparoscopic exploration with repair of all the defects. Prevention by carefully closing all potential mesenteric defects with running non-absorbable sutures during laparoscopic RYGBP, which we consider mandatory, seems appropriate in reducing the incidence of this complication. PMID- 17132416 TI - Cost-effectiveness and budget impact of obesity surgery in patients with type-2 diabetes in three European countries. AB - BACKGROUND: We aimed to establish a payer-perspective cost-effectiveness and budget impact (BI) model of adjustable gastric banding (AGB) and gastric bypass (GBP) vs conventional treatment (CT) in patients with BMI > or =35 kg/m(2) and type-2 diabetes T2DM, in Germany, UK and France. METHODS: Clinical evidence was obtained from literature and patient-reported EQ-5D scores given BMI and T2DM status from HODaR. Resource utilization data in AGB, GBP and CT were obtained from quoted publications so as to reflect practice in 2005. CT in each country was based on descriptions in HTA reports or based on co-authors' experience of current practice. Unit costs were obtained from published sources when available, or from co-authors' institutions. A deterministic algorithm with cost and utility discounting, enabled selection of inputs independently throughout the time scope for each of the 3 treatments, and included mean BMI, amounts of resources and unit costs. RESULTS: The base case time-scope was 5 years, and the annual discount rate for utilities and costs was 3.5%. Compared to CT, GBP yielded +80.8 kg/m(2).years, +2.6 T2DM-free-years and +1.34 QALYs. AGB yielded +57.8 kg/m(2).years, +2.5 T2DM-free-years and +1.03 QALYs. In Germany and France, both GBP and AGB yielded a cost decrease, and were thus dominant in terms of ICER compared to CT. In the UK, GBP and AGB yielded a cost increase, but were cost effective. CONCLUSION: In patients with T2DM and BMI > or =35 kg/m(2), AGB and GBP are effective at 5-year follow-up in cost-saving in Germany and France, and are cost-effective in the UK with a moderate BI vs CT. PMID- 17132417 TI - Use of PlasmaJet system in patients undergoing abdominal lipectomy following massive weight loss resulting from bariatric surgery: early experience. AB - BACKGROUND: Abdominoplasty has become a popular operation among patients seeking body contouring surgery due to the recent development of laparoscopic procedures in bariatric surgery and the epidemic diffusion of obesity. The wide surface of dissection is responsible for common postoperative complications such as seroma and hematoma. METHODS: PlasmaJet system (PJS), a high energy flow of ionized gas, can be used to stop capillary bleeding from blood and lymph vessels. We tested the PJS in a prospective series of 15 consecutive patients undergoing abdominoplasty after bariatric surgery-induced weight loss. RESULTS: 14 women underwent abdominoplasty with the PlasmaJet system after a mean weight loss of 48 kg (range 37-53). Mean operative time was 73 min (range 60-87). There was no postoperative complication. Mean fluid output from drains was 351.1 ml/patient (range 60 to 568), and drains were removed at a mean time of 4.8 days (range 3 to 6). CONCLUSION: These results are in favor of the efficacy of the PJS in reducing the amount of fluid production, and the rate of postoperative complications. However, this should be confirmed in a randomized trial comparing the PJS with standard technique. PMID- 17132418 TI - Long-term weight loss after bariatric surgery in patients visited at home outside the study environment. AB - BACKGROUND: Bariatric surgery results in sustained weight loss in the majority of patients. The controlled setting of a trial, however, does not necessarily reflect the everyday routine practice. Therefore, to investigate the results of bariatric surgery in a more natural setting, patients were visited at home, outside a study and hospital environment. METHODS: Patients who underwent a vertical banded gastroplasty or a gastric bypass between 1980 and 1997 were eligible. Body weight and height were measured. Patients filled out general and health status questionnaires (Nottingham Health Profile (NHP)). RESULTS: Responses were obtained from 236/313 subjects (75%) with a mean +/- SD age of 42.9 +/- 10.2 years and a mean current body weight of 100.3 +/- 20.7 kg. A maximum weight loss of 48.2 +/- 18.4 kg or 70.8 +/- 22.4% excess weight loss was obtained after 17 +/- 15 months, of which 32.1 +/- 22.6 kg or 45.2 +/- 29.3%, respectively, was maintained at 8.2 +/- 4.5 years after the intervention. Males and females did not differ in weight loss. The type of operation had no influence. Age >50 years and a BMI >50 kg/m(2) were not related to a poor outcome, but a time lapse of >5 years since the operation resulted in a less well sustained weight loss. The subjective health status improved considerably, but less so with a smaller weight loss and longer lapse of time since the operation. Especially in females, the NHP still deviated substantially from Dutch norm values. CONCLUSIONS: Surgically obtained weight loss is satisfactory in patients outside a strictly controlled study setting. Health benefits are substantial, but are adversely affected by weight gain and time elapsed since the operation. PMID- 17132419 TI - Changes in insulin sensitivity in morbidly obese patients with or without metabolic syndrome after gastric bypass. AB - BACKGROUND: We evaluated the medium term changes in insulin sensitivity in morbidly obese patients with and without metabolic syndrome before and after Roux en-Y gastric bypass (RYGBP) with silastic ring (Capella-Fobi). METHODS: A longitudinal, clinical intervention study was conducted in 40 patients between 18 and 65 years old, with obesity class II and III (BMI > or =35-52 kg/m(2)), divided into 2 groups: no metabolic syndrome (NMS, n=21) and metabolic syndrome (MS, n=19). Anthropometric measurements, biochemical tests and classification of MS according to the NCEP criteria, were performed pre-operatively and at 3 and 6 months postoperatively. RESULTS: In the preoperative period, 87% of the patients presented obesity class III (BMI 47+/-5 kg/m(2)) while 13% of the patients had obesity class II (37+/-2 kg/m(2)), and 19 patients (47.5%) presented MS. In the preoperative period, there were no differences among patients with MS and NMS in relation to the anthropometrics and body composition measurements. However, triglyceridemia, glycemia and insulinemia were higher in the MS group compared to the NMS group (P<0.05), although there was no difference in HOMA between the groups. HDL-cholesterol was lower in the MS group (p<0.05). In both postoperative study periods, all patients had significant reduction of anthropometric variables, body composition and biochemical variables. There were no differences between MS and NMS (p>0.05) groups. However, insulinemia decreased more in the postoperative period in the MS group compared to the NMS group (p<0.05). MS frequency in the MS group diminished to 26% after 3 postoperative months and no patient presented features of MS after 6 months postoperatively. CONCLUSIONS: Based on these observation: 1) patients of class II and III obesity present peripheral resistance to hyperinsulinemia without hyperglycemia; 2) RYGBP is able to reduce anthropometric measurements and body composition in a similar way for patients who have, or have not, MS; 3) there is rapid normalization of biochemistry of carbohydrates and lipids; 4) patients with previous MS lose the criteria needed for this diagnosis after 6 postoperative months. PMID- 17132420 TI - Effect of a high fat diet on quantitative features of adipocytes in the omentum: an experimental, stereological and ultrastructural study. AB - BACKGROUND: Omental adipose tissue specimens of female rats that were fed a high fat (HF) diet were evaluated stereologically and histopathologically. To our knowledge, there is no stereological study on numerical density, nuclear height and volume of adipocytes in omental adipose tissue in the female rat fed a HF diet in the literature. METHOD: 20 female Spraque Dawley rats were used in the study. 10 of the animals were fed HF diet consisting of 30% of calories from fat for 3 months. The remaining 10 rats, the control group, were fed a normal diet. After the experimental procedure, all animals were anesthetized and omental adipose tissues in the same area were dissected and fixed for the histochemical process using a mixture of 3% glutaraldehyde and 1% osmium tetraoxide in 0.1 M phosphate buffer. After embedding of tissues in araldite CY 212, semi-thin and thin sections were cut. The semi-thin sections were stained with toluidine blue. The physical dissector counting method was used for estimation of numerical density and nuclear height of adipocytes. Cavalieri principle was used for the estimation of adipocyte volume; volume fraction approach was applied to find the volume fraction of adipose tissue components. RESULTS: The mean numerical density of adipocytes in the HF diet group was significantly higher than the control. The mean nuclear height of adipocytes was also very high in the HF diet group. The volume fraction of adipose mass was increased whereas the extracellular matrix volume fraction was reduced in the HF diet group compared to the controls. The mean volume of adipocytes in the HF diet group was also significantly higher than in the control group. At the light microscopy level, it was found that adipocytes were enlarged and gaining irregular shape in the HF diet group. Thicker basal lamina and electron dense lipid content were also found in this group at the electron microscopy level. CONCLUSION: Lipid content and number of adipocytes in the adipose tissue of HF diet rats were higher than in the controls. Thus, HF diet induces increase in body weight via both hypertrophy and hyperplasia of adipocytes. PMID- 17132421 TI - Re-sleeve gastrectomy. AB - BACKGROUND: Laparoscopic sleeve gastrectomy (LSG) started as the restrictive part of the more complex laparoscopic duodenal switch (LDS) operation. There is no long-term experience with the isolated LSG. The main concern about the isolated LSG is the possibility of dilatation of the gastric pouch, long-term loss of restrictive function and weight regain. Laparoscopic re-sleeve gastrectomy (LRSG) has been used sparingly, but it also may become a possibility if more patients have the isolated LSG. METHODS: 2 patients with BMI 58 and 65 respectively, underwent LSG as the first stage of the LDS. Later, when the patients regained some weight and their gastric pouch was found to be too large, the LRSG/DS was done. RESULTS: The patient with BMI 58 had an initial drop to BMI 34 and regained weight to BMI 46, but after the LRSG/DS her BMI is 36 at 4 months. The BMI patient with BMI 65 had a drop to BMI 42, and after the LRSG/DS his BMI is 33 at 3 months later. CONCLUSION: LRSG may become necessary after gastric tube dilatation or insufficient original gastric volume reduction. LRSG is feasible, available and easy to perform when the resulting gastric pouch is too large or dilates after the original LSG. PMID- 17132422 TI - Laparoscopic latero-lateral jejuno-jejunostomy as a rescue procedure after complicated mini-gastric bypass. AB - Acute obstruction of jejunal limbs after gastric bypass surgery is rare but can result in a catastrophic scenario if the diagnosis is delayed. We report a 31 year-old female who developed acute efferent limb obstruction after a laparoscopic mini-gastric bypass (MGB), manifested as recurrent episodes of epigastric discomfort and bile-stained vomiting. The diagnosis was evident on oral contrast studies. She was successfully treated by a salvage laparoscopic side-to-side anastomosis between the efferent limb and the afferent limb 4 cm distal to the gastro-jejunostomy. Acute obstruction of the efferent limb after a MGB operation can be easily diagnosed and effectively treated by laparoscopic latero-lateral jejuno-jejunostomy. PMID- 17132423 TI - Gastric bypass for insulin resistance due to lipodystrophy. AB - We report a 41-year-old woman with severe insulin resistance due to partial lipodystrophy, who was successfully treated with gastric bypass surgery. PMID- 17132424 TI - Perforated ulcer at the gastrojejunostomy: laparoscopic repair after Roux-en-Y gastric bypass. AB - A 44-year-old woman was admitted from the emergency department with severe acute upper abdominal pain. The patient had undergone a laparoscopic Roux-en-Y gastric bypass (RYGBP) operation 16 months previously. CT scan showed intraabdominal free air. At emergency laparoscopic reoperation, a perforated ulcer at the gastrojejunostomy was found. This late complication of RYGBP can be a rapidly progressing life-threatening situation, and requires prompt treatment. Closure and omental patch were successful laparoscopically. PMID- 17132425 TI - Gastrogastric fistula after gastric bypass--is surgery always needed? AB - Because of regain of weight to BMI 37.1 kg/m(2) 6 years after a VBG, a 41-year old female now underwent revision to divided Roux-en-Y gastric bypass, performed laparoscopically. 12 days postoperatively, she started bleeding from the main stomach, and CT scan revealed that the bypassed stomach was distended with clot. She was treated conservatively and stopped bleeding. Upper GI series 2 weeks postoperatively revealed a large gastrogastric fistula between the tiny pouch and the bypassed stomach. We initially planned to close the fistula. However, upper GI series 2 months and 4 months postoperatively showed no sign of gastrogastric fistula, and proton pump inhibitors were stopped. At 1 year after gastric bypass, our patient has had good weight loss. PMID- 17132426 TI - Gastric cancer and Roux-en-Y gastric bypass. PMID- 17132427 TI - Update on punctal plugs. AB - Punctal plugs have offered a safe and often reversible treatment for aqueous deficient dry eye for over three decades. However, they represent only one tool in our armamentrarium to help patients with dry eyes, and plugs do have limitations. This article briefly reviews the history of occlusive treatments for aqueous tear deficiency and provides an update of recent advancements in punctal and canalicular occlusive materials and techniques. PMID- 17132429 TI - Sixth nerve palsy. AB - The diagnosis and etiologies of sixth cranial nerve palsies are reviewed along with nonsurgical and surgical treatment approaches. Surgical options depend on the function of the paretic muscle, the field of greatest symptoms, and the likelihood of inducing diplopia in additional fields by a given procedure. PMID- 17132431 TI - The neuro-ophthalmology of pituitary tumors. AB - The management of pituitary tumors requires a multidisciplinary approach involving neurosurgeons, endocrinologists, and ophthalmologists. Patients symptomatic from a pituitary tumor often are first evaluated by an ophthalmologist. This review discusses key issues regarding both the topographic diagnosis as well as the neuro-ophthalmologic manifestations of pituitary disease. Knowledge of the medical, surgical, and radiation treatment options for pituitary tumors is also important in coordinating patient care. The goals of such treatment include the control of tumor growth, normalization of pituitary function, and preservation or restoration of visual function. PMID- 17132433 TI - Refractive power of the cornea. AB - Corneal refractive surgeries, such as laser in situ keratomileusis and photorefractive keratectomy, have become some of the most commonly performed elective surgical procedures today. Many of the patients undergoing these surgeries are beginning to show signs of cataract formation and are in need of surgical correction. A common problem in the postrefractive patient is accurate prediction of the corneal power for use in intraocular lens calculation. The purpose of this article is to review the literature and to discuss why it is difficult to determine the power of the postkeratorefractive cornea, and to describe the multiple techniques used to assist in calculation of the power of the cornea. We will also examine some of the current technological advances that may aid in power calculation. With proper patient history, examination, and careful calculation(s), it is possible to closely estimate the refractive power of the postkeratorefractive cornea. PMID- 17132435 TI - Maculopathy in a child. PMID- 17132436 TI - The Eye Bank Association of America. PMID- 17132439 TI - Absorption spectrometry by narrowband light in optically saturated and optically pumped collision and Doppler broadened gaseous media under arbitrary optical thickness conditions. AB - This work examines absorption spectrometry by narrowband light in gaseous media with arbitrary optical thickness when the light induces optical saturation or optical pumping. Two quantities are defined: the observed absorbance, A(obs), and the true absorbance, A(true). The former is the absorbance that is measured under the existing conditions, whereas the latter represents the absorbance one would measure if the light acted solely as a probe of the populations of the various levels, and it is therefore directly proportional to the concentration or density of absorbers. A general integral equation for the propagation of light in media of arbitrary optical thickness in which the light influences the populations of the levels involved is derived. This expression is transcendental in the observed absorbance and cannot be solved analytically. It is shown that an analytical expression can be derived by investigating the inverse relationship, i.e., A(true) = f(A(obs)). Inasmuch as collision and Doppler broadened media react differently to optical saturation, they are considered separately. It is shown that a nonlinear response results if the medium is optically saturated (or pumped) and not optically thin. Expressions for the error introduced if the technique of standard additions is uncritically applied to such a system are derived. PMID- 17132440 TI - Fiber-optic surface plasmon resonance sensors in the near-infrared spectral region. AB - The sensitivity of fiber-optic surface plasmon resonance (SPR) sensors was improved by a factor of at least thirteen for aqueous solutions by modifying the tip geometry to allow interrogation of the surface plasmon (SP) band in the near infrared (NIR) region. This was achieved by tuning the angle at the distal end of the SPR sensor to a dual taper of 71 degrees and 19 degrees . Using a low numerical aperture (NA) fiber-optic sensor, NA = 0.12, is necessary to obtain a functional SPR sensor working in the NIR region. Theoretical simulations using the Maxwell equations demonstrated that even higher enhancement is theoretically possible while maintaining a narrow spectral feature upon the excitation of the SP bands on gold surfaces. The manufacture of the SPR sensors yields good agreement between theoretical simulations and experimental observations. To investigate the properties of these fiber-optic SPR-NIR sensors, sucrose solutions ranging from 0 to 15 x 10(-3) in mole fraction were utilized. The increased sensitivity of the fiber-optic SPR sensors, when used to monitor biomarkers, would yield lower detection limits. The smaller sensing area, compared to planar or other fiber-optic SPR sensors, combined with an improvement of the sensitivity, would yield a dramatic reduction of the absolute amount detected by biosensors. PMID- 17132441 TI - Drug characterization in low dosage pharmaceutical tablets using Raman microscopic mapping. AB - Raman micro-spectroscopic mapping is utilized to analyze pharmaceutical tablets containing a low concentration (0.5% w/w) of active pharmaceutical ingredient (API). The domain sizes and spatial distributions of the API and the major excipients are obtained. Domain size of the API is found to be dependent upon the particle size distribution of the ingoing API material, making the Raman maps good indicators of the source of API used in tablet manufacturing. Multivariate classification was performed to simultaneously check for the presence of two undesired API polymorphs within tablets. Raman mapping was demonstrated capable of detecting in the tablet matrix as little as 10% form conversion of the low dosage (0.5% w/w) API, which is equivalent to detection of a 0.05% w/w polymorphic impurity. Overall, the information provided by Raman micro spectroscopic mapping was found to have potential utility for manufacturing process optimization or predictive stability assessments. PMID- 17132442 TI - Attenuated total internal reflection infrared microspectroscopic imaging using a large-radius germanium internal reflection element and a linear array detector. AB - The number of techniques and instruments available for Fourier transform infrared (FT-IR) microspectroscopic imaging has grown significantly over the past few years. Attenuated total internal reflectance (ATR) FT-IR microspectroscopy reduces sample preparation time and has simplified the analysis of many difficult samples. FT-IR imaging has become a powerful analytical tool using either a focal plane array or a linear array detector, especially when coupled with a chemometric analysis package. The field of view of the ATR-IR microspectroscopic imaging area can be greatly increased from 300 x 300 microm to 2500 x 2500 microm using a larger internal reflection element of 12.5 mm radius instead of the typical 1.5 mm radius. This gives an area increase of 70x before aberrant effects become too great. Parameters evaluated include the change in penetration depth as a function of beam displacement, measurements of the active area, magnification factor, and change in spatial resolution over the imaging area. Drawbacks such as large file size will also be discussed. This technique has been successfully applied to the FT-IR imaging of polydimethylsiloxane foam cross-sections, latent human fingerprints, and a model inorganic mixture, which demonstrates the usefulness of the method for pharmaceuticals. PMID- 17132443 TI - Determination of soil nitrate and water content using attenuated total reflectance spectroscopy. AB - Direct determination of nitrate and soil moisture can significantly improve N application management and thus reduce N-derived environmental pollution related to agriculture. Several studies have shown that Fourier transform infrared attenuated total reflectance (FT-IR/ATR) spectroscopy could be used to estimate the nitrate content of standardized soil pastes. Paste standardization appeared to be the main obstacle to in situ application of this approach, and the present study shows how FT-IR/ATR can be used to estimate both water content and nitrate concentration of field soil samples. Water content and nitrate concentration are determined sequentially using two subsamples of the initial soil sample. An a priori determined amount of highly concentrated nitrate solution is added to the first subsample and the ATR spectrum of this paste is used to estimate the sample water content. It is then possible to calculate the amount of water that should be added to the second subsample so that the resulting paste is very close to the ideal standard paste. Nitrate concentration, mg [N]/kg [dry soil], is estimated using the FT-IR/ATR spectrum of this second paste. Results are presented for a laboratory experiment with four agricultural soils, as well as for a field trial with a calcareous soil. For water content, the determination errors range from 0.01 to 0.02 g [water]/g [dry soil]. For nitrate concentration, the errors for three of the soils range from 5.9 to 8.4 mg [N]/kg [dry soil], while for the fourth, calcareous clay soil, the determination error is 13.6 mg [N]/kg [dry soil]. The determination errors obtained for the field trial are similar to the ones obtained for a similar soil under laboratory conditions, which shows the potential usefulness of the approach for improving N-application management and reducing environmental pollution. PMID- 17132444 TI - Time-resolved Fourier transform infrared spectroscopy of chemical reactions in solution using a focal plane array detector. AB - A Fourier transform infrared (FT-IR) microscope equipped with a single as well as a 64 x 64 element focal plane array MCT detector was used to measure chemical reaction taking place in a microstructured flow cell designed for time-resolved FT-IR spectroscopy. The flow cell allows transmission measurements through aqueous solutions and incorporates a microstructured mixing unit. This unit achieves lamination of the two input streams with a cross-section of 300 x 5 microm each, resulting in fast diffusion-controlled mixing of the two input streams. Microscopic measurement at defined positions along the outlet channel allows time-resolved information of the reaction taking place in the flow cell to be obtained. In this paper we show experimental results on the model reaction between formaldehyde and sulfite. Using the single-point MCT detector, high quality FT-IR spectra could be obtained from a spot size of 80 x 200 microm whereas the FPA detector allowed recording light from an area of 260 x 260 microm focused on its 64 x 64 detector elements. Therefore, more closely spaced features could be discerned at the expense of a significantly lower signal-to-noise (S/N) ratio per spectrum. Multivariate curve resolution-alternating least squares was used to extract concentration profiles of the reacting species along the outlet channel axis. PMID- 17132446 TI - Photooxidative degradation and conformational changes in syndiotactic polypropylene/clay nanocomposites. AB - Syndiotactic polypropylene nanocomposites based on layered silicates in various proportions were subjected to prolonged (246 h) ultraviolet (UV)-c irradiation. Fourier transform infrared (FT-IR) spectroscopy was used in order to investigate the molecular alterations of the polymeric matrix during the UV exposure relative to the concentration of nanoclay. It was found that a significant increase of the helical conformation upon the irradiation took place as a result of scissions of the polymeric chains. In addition, a simultaneous increase in the crystallinity was verified by X-ray diffraction (XRD) measurements. Furthermore, a variety of photooxidation products were detected, among them carboxylic acids, ketones, gamma-lactones, and esters. We report in this paper the impact of the clay on the degradation mechanism of syndiotactic polypropylene mostly by the production of additional free radicals. Therefore, the relative intensities of the produced photooxidative species are affected drastically as a function of the concentration of the layered silicate present. Finally, an interaction of the carbonyl groups formed in the polymer upon irradiation with the polar groups of the layered silicates was confirmed both from infrared and UV-visible spectroscopic studies. PMID- 17132447 TI - Nondestructive identification of arsenic and cobalt minerals from Cobalt city, Ontario, Canada: arsenolite, erythrite, and spherocobaltite on pararammelsbergite. AB - A Ni-Co-As ore sample from Cobalt City, Ontario, Canada, was examined with scanning electron microscopy and energy dispersive X-ray analysis. In addition to cobaltian pararammelsbergite with variable cobalt content, for which Cobalt City is the type locality, and erythrite, one new mineral was observed for this locality. Well-formed crystals of arsenolite, As(2)O(3), were found embedded in what appears to be fibrous spherocobaltite, CoCO(3). Additional information was obtained by Raman microscopy, confirming the identification of the arsenolite. Both are considered to be secondary minerals formed by exposure to air resulting in oxidation and the formation of secondary carbonates. PMID- 17132448 TI - Application of Fourier transform Raman spectroscopy for prediction of bitterness of peptides. AB - The potential application of Fourier transform (FT) Raman spectroscopy to predict the bitterness of peptides was investigated. FT-Raman spectra were measured for the amino acid Phe and 9 synthetic di-, tri-, and tetra peptides composed of Phe, Gly, and Pro. Partial least squares regression (PLS)-1 analysis was applied to correlate the FT-Raman spectra with bitterness intensity values (R(caf) and log 1/T) reported in the literature. Using full cross-validation, Model 1 based on the single spectral data set for the nine peptides yielded a high correlation coefficient for calibration (R = 0.99), but a low correlation coefficient for prediction (R = 0.56). Two models were constructed using the data sets including replicate spectra for the calibrations and were validated using full cross validation. Using leave-one-sample-set-out calibrations, Model 2, which was developed with the data for the peptides as well as Phe, yielded a low correlation coefficient (R = 0.533) for the prediction of the bitterness, while Model 3 developed with only the peptide data provided better correlation coefficients (R = 0.807 and 0.724 for R(caf) and log 1/T values, respectively). The correlation coefficients for prediction were 0.975 (R(caf) values) and 0.874 (log 1/T values) for Model 4, which was developed using subtracted spectral data (spectra of peptides with higher R(caf) values minus spectra of peptides with lower R(caf) values). Examination of the PLS regression coefficients at wavenumbers most highly correlated with bitterness revealed the importance of hydrophobicity and peptide length on bitterness. This study indicates the potential of FT-Raman spectroscopy as a useful tool for predicting bitterness of peptides and amino acids. PMID- 17132449 TI - Spectrofluorimetric characterization of mixed nanoaggregates comprising a double chain cationic surfactant and a cationic or non-ionic single-chain surfactant. AB - A series of mixed vesicle and pre-vesicle nanoaggregates, comprising a cationic double-chain surfactant (di-decyldimethylammonium bromide, di-C(10)DMAB, or di dodecyldimethylammonium bromide, di-C(12)DMAB), and a cationic (dodecylethyldimethylammonium bromide, C(12)EDMAB) or non-ionic (octyl-beta-D glucopyranoside, OBG) single-chain surfactant have been characterized by means of steady-state fluorescence spectroscopy. For that purpose, the fluorescent emission of two probes, one anionic (TNS) and the other non-ionic (PRODAN), which is known to be sensitive to the polarity, rigidity, and/or microviscosity of the environment within which the probes are housed, has been measured in the presence of the above-mentioned mixed aggregates. The results of this analysis yield interesting information about the characteristics of the vesicle surface and bilayer, as well as about the existence of clusters and/or nanoaggregates prior to the formation of vesicles. PMID- 17132450 TI - Multivariate curve resolution of synchronous fluorescence spectra matrices of fulvic acids obtained as a function of pH. AB - Synchronous fluorescence spectra (excitation wavelength range between 280 and 510 nm and wavelength interval of 25 nm) of three samples of fulvic acids (FA) were obtained as a function of the pH, in the range from 2.0 to 10.5, and as a function of the FA concentration, in the range from 20 to 180 mg/L. FA were obtained from composted livestock materials (lsFA), composted sewage sludge (csFA), and Laurentian soil (laFA). Three-dimensional spectral matrices were obtained (wavelength, pH, and FA concentration) and multivariate curve resolution (MCR) was used to calculate spectra and fluorescence intensity profiles for the detected components. Cluster analysis of the calculated spectra showed the existence of similar and unique fluorescent properties in the three FA samples. Some of the calculated fluorescence intensity profiles have a shape compatible with acid-base species distribution diagrams, which allowed pKa values to be estimated, namely, a well-defined acid-base equilibrium with pKa 5.7 +/- 0.2 (lsFA), 6.9 +/- 0.4 (csFA), and 5.5 +/- 0.2 (laFA); and other acid-base systems not well defined with pKa at about 3.0 and 8.6. Other spectral variations revealed the existence of inner-filter effects or self-quenching as the concentration of FA increases. PMID- 17132451 TI - Detection and mapping of latent fingerprints by laser-induced breakdown spectroscopy. AB - Detection of latent fingerprints on a Si wafer by laser-induced breakdown spectroscopy (LIBS) is demonstrated using approximately 120 fs pulses at 400 nm with energies of 84 +/- 7 microJ. The presence of a fingerprint ridge is found by observing the Na emission lines from the transferred skin oil. The presence of the thin layer of transferred oil was also found to be sufficient to suppress the LIBS signal from the Si substrate, giving an alternative method of mapping the latent fingerprint using the Si emission. A two-dimensional image of a latent fingerprint can be successfully collected using these techniques. PMID- 17132452 TI - Fatty acid profiling of soybean cotyledons by near-infrared spectroscopy. AB - Genetically improved soybean grain often contains altered fatty acid profiles. Such alterations can have deleterious effects on seed germination and seedling development, making it necessary to monitor fatty acid profiles in follow-up physiological studies. The objective of this research was to quantify the five fatty acids in soybean (Glycine max) cotyledons using near-infrared (NIR) spectroscopy. Soybean cotyledon samples were dried, ground, and scanned with visible and NIR radiation from 400 to 2500 nm, and reflectance was recorded. Samples were also analyzed by gas chromatography (GC) for palmitic, stearic, oleic, linoleic, and linolenic acids and total oil; GC data, expressed as actual concentration and proportion of total oil, were regressed against spectral data to develop calibration equations. Equation statistics indicated that four of the five fatty acids could be predicted accurately by NIR spectroscopy; the fifth fatty acid could be determined by subtraction. Principal component analysis revealed that most of the spectral variation in this population was due to chlorophyll absorbance in the visible region. Therefore, the spectra were trimmed to include the NIR region only (1100-2500 nm), and a second set of equations was developed. Equations based exclusively on NIR spectra had equal or greater precision than equations based on visible and NIR spectra. Principal component analysis and partial least squares analysis revealed that even after trimming, at least 90% of the spectral variation was unrelated to fatty acid, though variation from fatty acid was identified in the second and third principal components. This research provides an NIR method for complete fatty acid profiling of soybean cotyledons. Equations were achieved with NIR spectra only, so spectrophotometers that analyze both the visible and NIR regions are not needed for this analysis. In addition, equations were possible with a 250 mg sample, which is one-tenth the normal sample size for this analysis. PMID- 17132453 TI - Spectroscopic assessment of Australian cotton waxes. AB - An investigation into the spectroscopic analysis of cotton waxes on Australian cottons was undertaken. The chemical composition of cotton wax is complex and contains a number of lipid classes. Infrared transmission spectroscopy coupled with principal component analysis was found to be capable of discriminating between solvent-extracted cotton waxes with differences in their alkyl functionality. Based on high-performance thin-layer chromatography (HPTLC) results, these differences were associated with an increase in levels of the alkane wax component. On the basis of these results, a photo-acoustic spectroscopic method was developed that could be used to distinguish raw cottons on the basis of these differences. This method was utilized to screen cottons from the Cotton Seed Distributors 2001 seed trial. A preliminary assessment of the scouring and dyeing properties of the various cottons, identified using the photo-acoustic method, was carried out. The results tended to confirm that cottons with increased alkyl functionality, most likely associated with alkane wax, were more difficult to remove and residual wax on the fiber acted as a barrier to dyestuff penetration, thus lowering color yield. PMID- 17132454 TI - Inverse spatially offset Raman spectroscopy for deep noninvasive probing of turbid media. AB - A new type of highly sensitive spatially offset Raman spectroscopy (SORS) developed for deep noninvasive probing of stratified turbid media is described. The technique, termed inverse SORS, permits much greater depths to be interrogated than those accessible with the conventional SORS approach. This is achieved by enhancing the sensitivity of the technique through the elimination of spectral distortions inherent to the conventional SORS methodology. The method also permits the use of higher laser powers in applications where intensity limits exist, such as when probing human tissue in vivo. In addition, the new approach possesses a much higher degree of flexibility, enabling on-the-spot tailoring of experimental conditions such as the magnitude and number of spatial offsets to individual samples. The scheme uses a reverse SORS geometry whereby Raman light is collected through fibers at the center of the probe and laser radiation is delivered to the sample through a beam in the shape of a ring. The method is demonstrated on a layered powder sample and several practical examples of its uses, presented for the first time, are also given. Potential applications include disease diagnosis, noninvasive probing of pharmaceutical products and chemicals through packaging, probing of polymers, biofilms or paints, and homeland security screening. PMID- 17132455 TI - Comments on the use of Raman spectroscopy for the in vivo measurement of human macular pigment. PMID- 17132456 TI - Lab tests online--an untapped resource. AB - Since its launch in 2001, Lab Tests Online has evolved into the go-to site for consumer information about laboratory testing. In some circles, however, it still remains underutilized as an educational tool for patients. This month, editor-in chief Anthony S. Kurec reviews the growth of Lab Tests Online and details its ongoing metamorphosis. PMID- 17132457 TI - Pro: benchmarking is the absolute prerequisite for timely and significant business process improvement. AB - Benchmarking in industry has been around for nearly a century, helping companies in nearly every sector imaginable improve their overall performance. Benchmarking's importance in health care, and specifically the clinical laboratory, can be summed up in one simple phrase--"If you cannot measure it, you cannot improve it." Here is why. PMID- 17132458 TI - Con: current laboratory benchmarking options are not good enough. AB - In an ideal world, benchmarking performance in the clinical laboratory would improve performance, quality, and overall patient satisfaction. However, there is a reason why laboratory managers continue to be on the lookout for the perfect benchmarking product--it doesn't exist. As a result, benchmarking performance in the laboratory is inherently flawed. Here is why. PMID- 17132459 TI - How to utilize benchmarking in the clinical laboratory. AB - Benchmarking of clinical laboratory activities has become a tool used increasingly to enable administrators and managers to obtain an independent evaluation of the performance of the laboratory and identify opportunities for improvement. Benchmarking is particularly important because of the diversity and complexity of the various sections of the laboratory. The critical component of laboratory benchmarking is peer comparison, as solutions to shortcomings or problems can be titrated and planned through this process. The reliability of benchmarking must be supplemented and modified by the input of the manager's detailed understanding of local circumstances. At this critical moment, the changes in peer review strategies instituted by JCAHO, CAP, CLIA, and individual states create an urgent opportunity to assist medical directors and laboratory managers in maintaining an overview of the performance and quality of laboratory operations. Unannounced site visits will require prompt reports and alerts of undesirable changes in performance. The future goals of benchmarking must expand to include surveys of laboratory test utilization and patient outcomes as ultimate measures of test utility in the clinical process and important assessments of the quality of patient care. PMID- 17132460 TI - Laboratory benchmarking: the College of American Pathologists' experience. AB - Benchmarking is an important part of performance evaluation in the clinical laboratory. When used effectively, benchmarking can lead to significant changes and performance improvement. This article reviews the experience of the College of American Pathologists (CAP) with benchmarking clinical laboratory expenses and presents data that summarizes recent laboratory trends identified through CAP's benchmark data. PMID- 17132461 TI - Setting up annual OSHA training--bloodborne pathogen standard. AB - While monitoring actual performance will always be of paramount importance, laboratory managers must also be vigilant about protecting their employees. In Part One of this two-part series discussing OSHA training procedures, columnist Judith O'Brien examines the necessary precautions related to bloodborne pathogens. PMID- 17132462 TI - Design your own laboratory disaster drills: a do-it-yourself guide. AB - You don't need to bring in a high-priced consultant to run a drill that gets your staff up to speed on emergency procedures. Follow along with Judy Bernice as she walks you through a step-by-step guide that will prepare your staff for any situation. PMID- 17132463 TI - There is no such thing as free pizza. AB - For many years, it has been standard practice in the United States to allow pharmaceutical representatives to provide drug samples, pens, note pads, visual aids, t-shirts, etc., and pay for attendee meals in conjunction with teaching conferences for hospital physicians. The "gifts" typically aren't as luxurious in the clinical laboratory, but even so, is any vendor freebie too much? PMID- 17132464 TI - [Times and spaces of care: home, hospital]. AB - In this article times and spaces of care offer the opportunity to reflect on nursing moving from an hermeneutic perspective. Our focus has been placed on what always undergo the tie that exists between every doing and the context in which it happens, rather that on the differences of practices in the various places and times of nursing activities--home care, hospital, the outpatients' department, and so on. According to the hermeneutic perspective, the outcome of nursing relation is not exclusively the re-solution of the nursing problem of the patient, but also--together with all of that--the possible experience of identity new construction (and re-birth) that every experience of disease represents. PMID- 17132465 TI - [The relative's need of participation in the care plan in a general medicine ward]. AB - All the scientific literature agrees on the fact that the shelter in hospital is a delicate moment for the patient. Also for relatives the shelter in hospital of their dear one is not of easy management, often they are excluded, insecure, alone and with a frankly uncertain role. The purpose of this study is to explore the role and the needs perceived from the relatives of an in-patient in a general medicine unit and to explore which role and which needs of the relatives are perceived from the nurses of a same ward. The sample of the study consisted of 49 relatives of in-patients in the ward of Medicina III dell'Azienda Ospedaliera di Reggio Emilia and of 18 nurses of the same ward. It was found that information is the most important need expressed by the relatives and that for the nurses is hard to satisfy it. Moreover, the nurses haven't a clear idea of the relative's role and they are inclined to exclude them from the care project. PMID- 17132466 TI - [Fatigue: cancer related problems and holistic approach]. AB - People suffering from cancer-related problems do not only need specific therapies, but also an adequate psychological support. Due to their specific function, nurses are in a position to comprehend these needs, whether they are expressed or not: this is a result not only of their technical and theoretical preparation, but also of their predisposition to a global approach to patients. This leads to a trusting relationship that allows the nurse to be supportive in dealing with one of the most frequent problems affecting cancer patients since better control of nausea and pain has been achieved: fatigue. For this purpose, research has begun in the Division of Medical Oncology Zone 3 of Fano composed of two steps: a cognitive survey and planning of suitable solutions. The results of the first step are described. PMID- 17132467 TI - [Nurses and burnout: a survey in an Emergency Department in the Lazio Region]. AB - Nowadays, burnout represents a growing phenomenon in technologically advanced countries, so much so that there is now abundant literature available on this topic. The aim of our study was to assess the level of burnout in a group of 108 nurses working in an Emergency Department by means of the Maslach Burnout Inventory. This instrument is based on the concept that burnout presents 3 basic features: Emotional Exhaustion (EE), Depersonalization (DP) and Personal Achievement (PA). Results showed that a significant relationship exists between EE and DP, the working environment and the same social-demographic features, while the PA sub-scale was not found to be dependent on any of the social demographic characteristics. PMID- 17132468 TI - [Female genital modification: survey of new phenomenon for nurses today]. AB - The immigration that there was in Italy in the last years has provoked a big cultural and social change for the country. Nurses that belong to social sector cannot escape from that change, and they have got to admit those new reality and develop new assistential and analysis methodologies. The immigrants that arrive in Italy came from more than 150 counties in the world, poor countries of the third world. The object of this research is to find out how nursing profession confronts itself with a new multiculture situation, the way to conciliate the ethical principles of the profession with the different cultures and learning that medical operators of an Italian hospital got about the immigrants: MGF (feminine genital modifications). The research wants to understand this phenomenon that is very diffuse, so the meeting with the differences is a reason to develop and not a imposition of a value on the other value. PMID- 17132469 TI - [Social-health operator: survey on the knowledges of nurses about this operator]. AB - The nurse is a full member of the multidisciplinary equipe and he is taken on the responsibility for operational correctness in the first person, acting am individually that in with the others sanitary operating collaboration and social and using, where necessary, of the support personnel, but when are cognizant of the responsibility from the attribution of an activity to a figure of support? Purpose of the search's that to understand that knowledge type had the nurses in worth to the figure of the operative sanitary partner and of the activity that compete him, and if were cognizant of the real responsibility on the attribution of the activity to this figures; besides the investigation contemplated to investigate if this knowledge/awareness they were influenced from the diverged working reality or from the share to courses of updating/formation. The approach has stated systematic, it has been considered a population composed by the nurses of three Italian hospitals. The surveys have stated effected across anonymous self-compiled questionnaires. From the results it is emersed that play a determinate role in the knowledge of the subject the frequented have updating courses and work in a working reality when this figure of support is present. PMID- 17132470 TI - The immune response of tilapia Oreochromis mossambicus and its susceptibility to Streptococcus iniae under stress in low and high temperatures. AB - Mozambique tilapia Oreochromis mossambicus acclimated to 27 degrees C were then held at 19, 23, 27 (control), 31 and 35 degrees C, and were examined for non specific cellular and humoral responses after 12-96 h. Total leucocyte count decreased significantly when fish were transferred to 19 and 23 degrees C after 48 and 96 h, and when transferred to 35 degrees C over 12-96 h, respectively. Respiratory burst decreased significantly when fish were transferred to 19, 31 and 35 degrees C over 24-96 h, whereas phagocytic activity and phagocytic index decreased significantly when fish were transferred to low temperatures (19 and 23 degrees C) and high temperatures (31 and 35 degrees C) over 12-96 h. Lysozyme activity decreased significantly when fish were transferred to 19 degrees C after 12-96 h, but increased significantly when transferred to 31 and 35 degrees C over 48-96 h. Alternative complement pathway (ACH(50)) also decreased significantly when transferred to 19 and 23 degrees C after 12h, but increased significantly when transferred to 31 and 35 degrees C after 24h. In another experiment, tilapia reared at 27 degrees C were injected intraperitoneally with Streptococcus iniae at a dose of 1 x 10(7)colony-forming units (cfu)fish(-1), and then reared onward at water temperatures of 19, 23, 27 (control), 31 and 35 degrees C. Over 48-168 h, the cumulative mortality of S. iniae-injected fish held in 19 and 35 degrees C was significantly higher than that of injected-fish held in 23, 27 and 31 degrees C. It is concluded that transfer of tilapia O. mossambicus from 27 degrees C to low temperatures (19 and 23 degrees C) after 12h, and transfer of fish from 27 degrees C to high temperatures (31 and 35 degrees C) reduced their immune capability. Moreover, tilapia under temperature stress at 19 and 35 degrees C from 27 degrees C decreased its resistance against S. iniae. PMID- 17132471 TI - Stress and immune response in the mussel Mytilus galloprovincialis. AB - The present study investigates the effects on immune-related parameters of various stress factors (air exposure, mechanical stress, high temperature and extreme salinity conditions) faced by the bivalve mollusc Mytilus galloprovincialis during marketing procedures. We observed that some stress typologies increase phagocytosis and the number of circulating immunocytes, while others can modify immunocyte response towards a further perturbation, i.e. the marine algal toxin yessotoxin. Our results suggest that non-lethal stress can be counteracted for sometime by increasing the level of some defence parameters. Moreover, our data indicate that fishing and transport procedures could interfere with mussel immunosurveillance. PMID- 17132472 TI - The integration of genomics into obstetrics and gynecology: a HuGE challenge. PMID- 17132473 TI - Preterm labor is induced by intraamniotic infusions of interleukin-1beta and tumor necrosis factor-alpha but not by interleukin-6 or interleukin-8 in a nonhuman primate model. AB - OBJECTIVES: The purpose of this study was to determine the relative contributions of individual proinflammatory cytokines and chemokines to the triggering of preterm labor. STUDY DESIGN: Eighteen chronically instrumented pregnant rhesus monkeys at 135 +/- 3 days gestation (term = 167 days) received 1 of 5 intraamniotic infusions: (1) interleukin-1beta (IL-1beta) (10 microg; n = 5), (2) tumor necrosis factor-alpha (TNF-alpha) (10-100 microg; n = 5), (3) IL-6 (20 microg twice a day; n = 2), (4) IL-8 (20 microg twice a day; n = 2), and (5) saline control (n = 4). Primary study outcomes were the mean uterine hourly contraction area (mm Hg x s/h) in 24 hours during peak response to cytokine infusion (all groups) and the interval from cytokine infusion until labor onset (IL-1beta, IL-6, and IL-8 groups). Secondary outcomes were quantities of amniotic fluid cytokines and chemokines (IL-1beta, TNF-alpha, IL-6, and IL-8), prostaglandins E2 and F2alpha, leukocytes, and matrix metalloproteinase-9 (MMP 9). Histopathology of fetal lungs and placental membranes was assessed. RESULTS: IL-1beta stimulated the most intense contraction patterns, resulting in preterm labor in all cases. TNF-alpha induced a variable degree of uterine activity among individual animals stimulating either preterm labor (n = 2) or a uterine contraction pattern of moderate intensity (n = 3). Despite prolonged elevations in amniotic fluid levels, neither IL-6 nor IL-8 induced preterm labor or an increase in uterine activity until near term. The mean interval from the initiation of IL-6 and IL-8 infusion to the onset of labor was significantly longer than after IL-1beta (21.9 vs 1.1 days; P < .01), and did not differ from the saline control group (27.6 days; P = NS). Intraamniotic infusion of IL-1beta or TNF-alpha was associated with significant elevations in all tested amniotic fluid cytokines, IL-8, prostaglandins, MMP-9 and leukocytes compared with gestational age-matched saline controls. IL-6 and IL-8 infusions were not associated with increases in IL-1beta or TNF-alpha and only produced a moderate increase in amniotic fluid prostaglandins. All cytokine infusions induced histologic chorioamnionitis and an accumulation of neutrophils in fetal lungs. CONCLUSION: Preterm labor was induced by intraamniotic infusions of IL-1beta and TNF-alpha, but not by IL-6 or IL-8 although inflammatory changes in fetal membranes and lungs were uniformly present. Our results indicate a primary role for IL-1beta and TNF-alpha in the triggering of preterm labor associated with inflammation or infection. PMID- 17132474 TI - The impact of the overactive bladder syndrome on sexual function: a preliminary report from the Multicenter Assessment of Transdermal Therapy in Overactive Bladder with Oxybutynin trial. AB - OBJECTIVE: The objective of the study was to determine the impact of overactive bladder on sexual function from a preliminary analysis of the Multicenter Assessment of Transdermal Therapy in Overactive Bladder with Oxybutynin study. STUDY DESIGN: The Multicenter Assessment of Transdermal Therapy in Overactive Bladder with Oxybutynin study was an open-label, prospective trial of 2878 subjects with overactive bladder, treated with transdermal oxybutynin for 6 months or less. The impact of overactive bladder on sexual function before and after treatment was assessed via item responses from the King's Health Questionnaire and Beck Depression Inventory-II (kappa-test). RESULTS: At baseline, 586 (23.1%) reported that overactive bladder had an impact on their sex life. Coital incontinence in 569 (22.8%) decreased after treatment to 438 (19.3%). Effects of overactive bladder on subjects' sex lives improved in 19.1% (worsened in 11.2%), and the effect on relationships with partners improved in 19.6% (worsened in 11.9%). Reduced interest in sex, reported by 52.1% at baseline, improved significantly. (all P < .0001). CONCLUSION: Overactive bladder negatively affects sexual function. Treatment with transdermal oxybutynin improved sexual function and marital relationships. PMID- 17132475 TI - Vascular anatomy of the presacral space in unembalmed female cadavers. AB - OBJECTIVE: The purpose of this study was to characterize the vascular anatomy of the female presacral space (PSS) and to correlate findings to the abdominal sacrocolpopexy. STUDY DESIGN: Detailed dissections of the PSS were performed in 52 unembalmed female cadavers. RESULTS: The closest cephalad vessel to the mid sacral promontory (MSP) was the left common iliac vein (LCIV), mean distance 27 mm (9-52 mm). The closest vessel lateral to MSP was also the LCIV, mean distance 22 mm (9-35 mm). The average distance of the middle sacral artery and vein to the MSP was 4 mm (0-15 mm) and 7 mm (0-17 mm), respectively. The mean distance of the sacral venous plexuses to the MSP was 34 mm (4-86 mm). CONCLUSION: Anatomic location of the vascular boundaries and contents of the PSS is highly variable. Careful dissection and exposure of the anterior longitudinal ligament of the sacrum before suture placement should minimize potentially life-threatening vascular complications. PMID- 17132476 TI - Posterior vaginal wall prolapse does not correlate with fecal symptoms or objective measures of anorectal function. AB - OBJECTIVE: The purpose of this study was to evaluate the relationship among the degree of posterior vaginal wall prolapse, anorectal symptoms, and physiology. STUDY DESIGN: This was a prospective study that included patients with fecal dysfunction and prolapse/urinary symptoms. A validated instrument for fecal incontinence and the ROME II criteria were used. Anal physiologic testing was performed selectively on the basis of bowel symptoms. Patients were divided into 2 groups by pelvic organ prolapse quantification score clinical examination: group I: Ap/Bp < -1, and group II: Ap/Bp > or = -1. The association among the degree of prolapse, bowel symptoms, and physiologic findings was examined with the use of Fisher's exact test and logistic regression models. RESULTS: One hundred thirty-two patients with a mean age of 63 years (range, 24-90 years) were evaluated. There were 62 patients (47%) in group I and 70 patients (53%) in group II. Overall, 40.9% of the patients had constipation/obstructed defecation, and 25% of the patients had fecal incontinence. Ninety-seven patients underwent physiologic testing. At manometry, both resting and squeeze pressures were significantly higher in patients in group II. Overall, patients with a higher resting pressure (P = .001) and increased rectal capacity (P = .008) were more likely to be continent, and patients with a lower squeeze pressure were more likely to be incontinent (P = .001). Ultrasonography demonstrated anterior sphincter defect in 21.9% of the patients and a perineal body of <10 mm in 35.6% of the patients, with no correlation with the degree of prolapse. Patients with sphincter defects were 3 times more likely to have fecal incontinence (95% CI, 1.03, 8.75; P = .04). There was no association between electromyography and pudendal nerve terminal latencies and the degree of prolapse. Patients with prolonged pudendal nerve terminal latencies were more likely to be incontinent (P = .033). On defecography, 28% of the patients had concomitant enterocele, and 21.3% of the patients had intussusception, which was not detected by physical examination. CONCLUSION: Anorectal symptoms do not correlate with the degree of posterior vaginal wall prolapse, nor does the presence of prolapse equate to abnormal physiologic test results. Bowel symptoms may result from primary anorectal abnormalities, which are demonstrated by physiologic studies. PMID- 17132477 TI - Repair of obstetric vesicovaginal fistulas in Africa. AB - OBJECTIVE: The purpose of this study was to show the outcomes of primary surgical repair of obstetric vesicovaginal fistula repairs in Niger, Africa. STUDY DESIGN: From October 2003 to April 2005, 90 patients were examined with vesicovaginal fistulas and no previous repair. Fistulas were variable in location and degree of scarring and ranged up to 7 cm in size. All patients were offered surgical repair. Primary repair was performed in 73 women. RESULTS: Successful primary closure was achieved in 41 patients (56%); 19 patients were lost to follow up, and 13 patients had a persistent fistula. Incontinence, despite fistula closure, was present in 9 patients. Common features of failure and/or incontinence included larger size, involvement of the urethrovesical junction, and scarring. CONCLUSION: The initial repair of vesicovaginal fistulas has the highest probability of success. The successful closure is dependent on size, site, and associated scarring. PMID- 17132478 TI - Sexual function, quality of life, and severity of anal incontinence after anal sphincteroplasty. AB - OBJECTIVE: The purpose of this study was to determine the severity of anal incontinence and its impact on quality of life and sexual function in women after anal sphincteroplasty. STUDY DESIGN: Eighty-six women who underwent anal sphincteroplasty during the years 1993 to 2004 were mailed validated survey instruments to evaluate continence status, health-related quality of life, and sexual functioning. Demographic and perioperative data were obtained from patient charts. RESULTS: At a mean follow-up time of 5.6 +/- 3.0 years, 6 women (11%) were totally continent; 8 women (15%) were incontinent of flatus only, and 41 women (75%) were incontinent of liquid and/or solid stool. Sexual function scores were not correlated with continence scores; 24% vs 4% of subjects who had undergone an overlapping sphincteroplasty versus an end-to-end sphincteroplasty reported pain during intercourse (P = .04). CONCLUSION: Anal continence rates 5 years after anal sphincteroplasty are disappointing, adversely impact quality of life, yet do not appear to relate to sexual function. PMID- 17132479 TI - Rectocele repair: a randomized trial of three surgical techniques including graft augmentation. AB - OBJECTIVE: This study was undertaken to compare outcomes of 3 different rectocele repair techniques. STUDY DESIGN: One hundred six women with stage II or greater posterior vaginal wall prolapse were randomly assigned to either posterior colporrhaphy (n = 37), site-specific rectocele repair (n = 37), or site-specific rectocele repair augmented with a porcine small intestinal submucosa graft (Fortagen, Organogenesis, Inc, Canton, MA; n = 32). Subjects underwent a physical examination and completed 3 validated pelvic floor instruments at baseline and 6 months, 1 year, and 2 years after surgery. Anatomic failure was defined as pelvic organ prolapse quantitation system (POPQ) point Bp > or = -2 at 1 year. RESULTS: Of 106 subjects who enrolled, 105 underwent surgery and of those 105, 98 subjects returned (93%) with a mean follow-up of 17.5 +/- 7 months. After 1 year, those subjects who received graft augmentation had a significantly greater anatomic failure rate (12/26; 46%) than those who received site-specific repair alone (6/27; 22%) or posterior colporraphy (4/28; 14%), P = .02. There was a significant improvement in prolapse and colorectal scales and overall summary scores of the Pelvic Floor Distress Inventory short form 20 (PFDI-20), the Pelvic Floor Impact Questionnaire short form 7 (PFIQ-7) after surgery in all groups (P < .001 for each) with no differences between groups. The proportion of subjects with functional failures was 15% overall, and not significantly different between groups. There was no significant change in the rate of dyspareunia 1 year after surgery and there were no differences between groups. Overall sexual function as measured by the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire short form (PISQ-12) improved significantly in all groups postoperatively (P < . 001), with no differences between groups. CONCLUSION: Posterior colporraphy and site-specific rectocele repair result in similar anatomic and functional outcomes. The addition of a porcine-derived graft does not improve anatomic outcomes. All 3 methods of rectocele repair result in significant improvements in symptoms, quality of life, and sexual function. PMID- 17132480 TI - Glycodelin mediates the increase in vascular endothelial growth factor in response to oxidative stress in the endometrium. AB - OBJECTIVE: We hypothesized that glycodelin stimulates vascular endothelial growth factor (VEGF) expression in response to oxidative stress. STUDY DESIGN: EM42 (human endometrial epithelial cell line) and primary endometrial epithelial cells were subjected to oxidative stress with minimally oxidized low density lipoprotein (mLDL). Cells were also incubated with no LDL (control) or native LDL (nLDL). Each condition was incubated with and without glycodelin antibody. Glycodelin and VEGF protein and messenger RNA (mRNA) levels were analyzed. Primary cells were cultured with glycodelin peptide to evaluate the effect on VEGF protein and mRNA. RESULTS: Glycodelin and VEGF protein and mRNA were higher for cells grown with mLDL (P < .05), while glycodelin antibody attenuated the increase in VEGF protein (P < .01). Glycodelin peptide increased VEGF mRNA and protein (P < .05). CONCLUSION: Glycodelin may act as an autocrine factor within endometriotic implants to increase VEGF expression during oxidative stress. PMID- 17132481 TI - Quality of life and continence 1 year after the tension-free vaginal tape operation. AB - OBJECTIVE: The purpose of this study was to evaluate changes in condition specific and generic quality of life (QOL) and continence rates 1 year after the tension-free vaginal tape (TVT) operation for stress urinary incontinence. STUDY DESIGN: A total of 99 patients undergoing the TVT operation underwent clinical and urodynamic assessment and completed the German language King's Health Questionnaire (KHQ) and the Short Form 36 (SF-36) before and 3 and 12 months after surgery. Overall, 72 TVTs were performed as isolated procedure and 27 in combination with other operations. Data were analyzed with the Chi-square test, analyses of variance, and the Tukey test. RESULTS: Overall, the objective continence rate was 80% at 1 year and did not differ significantly between patients with or without concomitant surgery. Significant and clinically relevant (>10 points) improvements were seen in all domains of the KHQ except General Health perception. The SF-36 showed statistically significant but not clinically significant improvement. CONCLUSION: The TVT operation is associated with improved condition-specific QOL in women with stress urinary incontinence. The condition-specific KHQ is more appropriate than the generic SF-36 for evaluating treatment results in women treated for incontinence. PMID- 17132482 TI - Clinical anatomy and surgical skills training (CASST): development of a multicenter, multidisciplinary program. AB - OBJECTIVE: The aim of this program was to develop a multicenter, multidisciplinary anatomy and surgical skills training program for junior residents in obstetrics and gynecology and urology. STUDY DESIGN: After administering a needs assessment, we developed a collaborative clinical anatomy and surgical skills training program for junior residents in obstetrics and gynecology and urology at 3 academic medical centers in Chicago. RESULTS: Thirty two residents participated in the program. Needs assessment results indicated that all residents felt they could benefit by more formal training in basic surgical skills. Learning objectives were developed for each of the 5 3-hour sessions that dealt with basic surgical skills, anterior abdominal wall anatomy, opening and closing the abdomen, female pelvic anatomy, and perineal anatomy and laceration repair. The cost of training each of the residents was approximately 600 dollars. Forty-five percent of the costs were one-time "start-up" costs for abdominal trainers and surgical instruments. CONCLUSION: By including multiple centers and disciplines, we were able to reduce costs of teaching basic surgical skills and anatomy and maximize faculty teaching time and effort. PMID- 17132483 TI - A randomized trial comparing methods of vaginal cuff closure at vaginal hysterectomy and the effect on vaginal length. AB - OBJECTIVE: The objective of the study was to compare the effect of horizontal versus vertical closure of the vaginal cuff during vaginal hysterectomy on vaginal length. STUDY DESIGN: Forty-three women were randomized to undergo horizontal (n = 23) or vertical (n = 20) vaginal cuff closure during vaginal hysterectomy at a community hospital. The primary outcome of vaginal length before and after surgery was compared by the Student t test and the paired t test. RESULTS: Preoperatively mean vaginal lengths in the horizontal and vertical groups were statistically similar (7.76 +/- 1.23 cm versus 8.28 +/- 1.39 cm, respectively; P = .21). Postoperatively the groups statistically differed (6.63 +/- 1.02 cm versus 7.93 +/- 1.18 cm, P < .001). The mean change in vaginal length was -1.13 +/- 1.15 cm and -0.35 +/- 0.91 cm, respectively (P = .01). Within-group comparisons revealed a statistical difference between pre- versus postmean vaginal length in the horizontal group (7.76 +/- 1.23 cm versus 6.63 +/- 1.02 cm; P < .001) and no difference within the vertical group (8.28 +/- 1.39 cm versus 7.93 +/- 1.18 cm; P = .11). CONCLUSION: Closing the vaginal cuff vertically is superior to horizontal closure for the purpose of preserving vaginal length. PMID- 17132484 TI - Anatomical path of the tension-free vaginal tape: reassessing current teachings. AB - OBJECTIVE: The objective of the study was to revisit the anatomical path of the tension-free vaginal tape and better describe its relationship to the perineal membrane and other important anatomic landmarks. STUDY DESIGN: Dissections of the anterior perineal triangle, periurethral, and retropubic spaces were performed in 24 unembalmed female cadavers following placement of the tension-free vaginal tape to identify the sling's relationship to the perineal membrane, periurethral muscles, and the arcus tendineus fascia pelvis. RESULTS: In 100% of specimens, the device passed cephalad to the perineal membrane. The urethrovaginal sphincter muscle was perforated in 2 of the specimens. The sling passed lateral to the arcus tendineus and perforated the pubococcygeus muscle in 6 (25%) of the cadavers. In the remaining 18 (75%) specimens, the mesh was medial to the arcus tendineus and penetrated the periurethral connective tissue. CONCLUSION: The assertion that the tension-free vaginal tape perforates the perineal membrane is incorrect. PMID- 17132485 TI - Advanced anterior vaginal wall prolapse is highly correlated with apical prolapse. AB - OBJECTIVE: The purpose of this study was to determine the relationship between the most prolapsed portion of the anterior and posterior vaginal walls and the apex. STUDY DESIGN: After obtaining institutional review board approval, demographic data and pelvic organ prolapse quantification findings from consecutive new patients who were seen at our urogynecologic practice between January 2004 and February 2005 were reviewed. RESULTS: Three hundred twenty-five women were included in this cohort. The support of the vaginal apex (pelvic organ prolapse quantification point C) correlated strongly with the most prolapsed portion of the anterior vaginal wall, Ba (Spearman's rho = 0.835; P < .001) and correlated moderately with the most prolapsed portion of the posterior vaginal wall, Bp (Spearman's rho = 0.556; P < .001). A strong linear relationship was found between C and Ba, which is best modeled by the following regression equation: C = Ba(1.4) - 4.4 (r = .869). CONCLUSION: Anterior vaginal wall prolapse is associated strongly with apical prolapse. Anterior vaginal wall defects that are surgically repaired usually require a concomitant repair of the apex. PMID- 17132486 TI - Neuropathic injury to the levator ani occurs in 1 in 4 primiparous women. AB - OBJECTIVE: We measured levator ani neuromuscular function before and after first delivery to identify the location, timing, and mechanism of injury. STUDY DESIGN: Fifty-eight primiparous women underwent electromyographic examination of the levator ani antepartum at 6 weeks and 6 months after the delivery. Antepartum turns/amplitude data were pooled to create a normal range. We calculated each woman's percentage of outliers from this range and assessed relationships between delivery and extent of injury. RESULTS: At 6 weeks, 14 of 58 women (24.1%) had neuropathy, with 9 of those 14 women recovering by 6 months. At 6 months, 17 of 58 women (29.3%) were neuropathic, which included 12 new injuries. Women who had elective cesarean delivery had virtually no injury, but all other modes of delivery had similar injury rates. CONCLUSION: Obstetric delivery is associated frequently with electromyographic evidence of neuropathic injury to the levator ani. The entire levator complex is at risk, and cesarean delivery while in labor is not protective. PMID- 17132487 TI - Outcome of tension-free vaginal tape procedure when complicated by intraoperative cystotomy. AB - OBJECTIVE: The objective of this study was to determine risk factors for intra operative cystotomy during TVT and whether this affects surgical outcome and morbidity. STUDY DESIGN: Charts of 340 women who underwent TVT were reviewed. Those who had a cystotomy during the procedure (cases) were compared with those without (controls) for the variables: age, race, parity, body mass index (BMI), TVT with or without other procedures, previous pelvic surgery, blood loss, length of catheter drainage, UTI, and voiding dysfunction. The t-test and chi-squared test were used and differences resulting in P < .05 were statistically significant. RESULTS: Cystotomy occurred in 49/340 TVT procedures (14.4%). Only those who had a history of abdominal hysterectomy and BMI greater than 26.5 were more likely to have a cystotomy (P = .05 and P = .001, respectively). Cases were more likely to go home with catheter drainage (P = .005). There was no increased risk of UTI or voiding dysfunction, nor was there a difference in surgical cure rate. CONCLUSION: Cystotomy does not seem to negatively affect the outcome of the TVT procedure. PMID- 17132488 TI - Role of lymphadenectomy in the management of grossly apparent advanced stage epithelial ovarian cancer. AB - OBJECTIVE: The purpose of this study was to determine the factors that are related to the performance of lymph node assessment and its impact on prognosis in ovarian cancer. STUDY DESIGN: This was a retrospective analysis of stage IIIC/IV epithelial ovarian cancer in patients who had undergone primary surgery between 1994 and 1998. Simple statistics and univariate and multivariable analysis were performed. RESULTS: Two hundred nineteen patients met the inclusion criteria; lymph node assessment was performed for 93 of these patients (41%). Sixty-one patients (65.5%) underwent complete pelvic and para-aortic lymphadenectomy, and 32 patients (34.5%) underwent a more limited lymph node sampling. In patients with residual disease >1 cm, lymph node assessment was an independent predictor of outcome. In this same subgroup, lymphadenectomy appeared to be superior to lymph node sampling (5-year overall survival, 50% (lymphadenectomy) vs 33% (lymph node sampling) vs 29% (no lymph node assessment); P = .01). Considering survival of the subgroup who underwent lymph node assessment, we observed a significantly worse outcome for those with lymphatic involvement (5-year overall survival, 31.5% [positive for nodal metastases] vs 54% [negative for nodal metastases]; P = .003). Although multiple factors were correlated with the decision to perform lymph node assessment in univariate analysis, only the surgeon (P < .001), low residual disease (P = .004), American Society of Anesthesiology 1 or 2 (P = .004), and the absence of carcinomatosis (P = .0002) were independent factors in the multivariable analysis. Further, if lymph node assessment was performed, the decision to do lymphadenectomy versus lymph node sampling was associated independently with the surgeon (P < .001), low residual disease (P < .001), and patient age of <65 years (P < .001). CONCLUSION: Removal of obviously involved lymph nodes in patients with residual disease near 1 cm and lymphadenectomy for patients with complete or near complete resection of abdominal disease appears to be justified. A lack of standard recommendation in advanced ovarian cancer results in wide variations that are based on individual preference in addition to logical factors. PMID- 17132489 TI - Interpretation of the Term Breech Trial findings. PMID- 17132490 TI - Extirpative or conservative management for placenta percreta? PMID- 17132493 TI - Cytokine sensitivity and N-glycan processing mutations. AB - The EGF and TGF-beta families of cytokines are critical regulators of cell proliferation, morphogenesis, and tissue repair. The signaling pathways downstream of EGF and TGF-beta receptors also contribute to cancer growth and metastasis. Cytokine receptors are glycoproteins, and we have recently shown that GlcNAc-branching of N-glycans enhances their cell surface residency and contributes to the growth autonomy of cancer cells. Ligand-induced dimerization of EGF receptors leads to phosphorylation of Erk1/2, whereas TGF-beta binding to its receptors stimulates phosphorylation of Smad2/3. Activated Erk1/2 and Smad2/3 translocate independently into the nucleus and regulate gene expression. Here we describe a sensitive and robust method to quantify TGF-beta and EGF signaling in cancer cells and primary cells from mice by quantitative fluorescence imaging. PMID- 17132494 TI - Phenotype changes of Fut8 knockout mouse: core fucosylation is crucial for the function of growth factor receptor(s). AB - Alpha1,6-fucosyltransferase (Fut8) catalyzes the transfer of a fucose residue to N-linked oligosaccharides on glycoproteins by means of an alpha1,6-linkage to form core fucosylation in mammals. In mice, disruption of Fut8 induces severe growth retardation, early death during postnatal development, and emphysema-like changes in the lung. A marked dysregulation of TGF-beta1 receptor activation and signaling in Fut8-null mice lung results in overexpression of matrix metalloproteinases (MMPs), such as MMP12 and MMP13, and a down-regulation of extracellular matrix (ECM) proteins such as elastin, which contributes to the destructive emphysema-like phenotype observed in Fut8-null mice. Furthermore, therapeutic administration of exogenous TGF-beta1 rescued the null mice from the emphysema-like phenotype. On the other hand, absence of Fut8 on EGF or PDGF receptor results in down-regulation of the receptor-mediated signaling, which is a plausible factor that may be responsible for the growth retardation. Reintroduction of the Fut8 gene to Fut8-null cells potentially rescued these receptor-mediated signaling impaired in null cells. Collectively, these results suggest that core fucosylation is crucial for growth factor receptors such as TGF beta1 and EGF receptor-mediated biological functions. PMID- 17132495 TI - Cellular and molecular analysis of neural development of glycosyltransferase gene knockout mice. AB - Recent studies demonstrate that carbohydrates synthesized by specific glycosyltransferases play important roles in the development of the central nervous system. Among these carbohydrates, polysialic acid is a unique glycan that modulates functions of the neural cell adhesion molecule (NCAM) by attenuating NCAM-mediated interaction between neural cells. During brain development, polysialic acid is synthesized in a specific spatiotemporal pattern by two polysialyltransferases, ST8SiaII and ST8SiaIV. To study in vivo the roles of polysialic acid synthesized by each respective enzyme, we generated ST8SiaII and ST8SiaIV knockout mice. Single knockout ST8SiaII or ST8SiaIV mice show polysialic acid expression patterns differing from wild type, and those patterns indicate different roles of each gene during neural development. In this chapter, we discuss methods used to analyze polysialyltransferase knockout mice using immunohistochemistry of brain and primary cultures of neurons. PMID- 17132496 TI - Roles of glycolipids in the development and maintenance of nervous tissues. AB - Glycoshingolipids are involved in a wide variety of biological events, including cell proliferation, differentiation, development, regeneration, and apoptosis in vertebrates. Expression profiles of glycolipids during the development and cell differentiation or transformation suggest that glycolipids are largely implicated in the determination of cell fates by directly transducing biosignals as receptors and/or modulating receptors' function. Despite of a number of efforts to clarify the molecular functions of glycolipids, no unambiguous results have been obtained until genetic modification of glycolipids became possible. Recent progress in the isolation of cDNAs of glycosphingolipid synthase genes has enabled us to examine roles of glycosphingolipids and strongly promoted further understanding of significances of glycosphingolipids. In particular, knock-out mice of glycosyltransferases showed quite novel aspects of glycolipid function and also redundancy among similar enzymes and glycolipid structures. Here, we summarize analytical methods with which roles of glycolipids in the development and maintenance of nervous tissues, including techniques to establish transgenic mice and gene knock-out mice, to survey fundamental behavior abnormalities, and to examine fine morphological changes lying under abnormal phenotypes of the glycolipids-modified cells and glycolipid-lacking mutant mice. PMID- 17132497 TI - Analysis of neural cell functions in gene knockout mice: electrophysiological investigation of synaptic plasticity in acute hippocampal slices. AB - Several knockout mice deficient in transferases, required for glycosylation of cell adhesion and extracellular matrix molecules, have recently been produced. Extracellular recordings of field excitatory postsynaptic potentials in acute hippocampal slices prepared from these mutant mice proved to be a highly sensitive method to reveal the roles of transferases and related carbohydrates in synaptic transmission and plasticity. Although most available data have been collected for synaptic connections between CA3 and CA1 pyramidal cells, several other synapses are assessable for extracellular recording in the hippocampus, including connections between mossy fibers and CA3 pyramidal cells. Analysis of distinct forms of short- and long-term plasticity in these connections may be instrumental for dissection of mechanisms by which carbohydrates affect synaptic functions. PMID- 17132498 TI - Analysis of neural cell function in gene knockout mice: behavior. AB - A powerful tool to investigative gene function is the ability to create mice with targeted gene mutations. Analysis of the resulting phenotype is sometimes difficult, however, because individual genes have more than one function, and observed effects on complex behaviors are often a result of abnormalities of any of a number of individual processes. One way to address this issue is by examining mice in a battery of behavioral tests to assess the specificity of any observed differences among genotypes. This chapter describes a test battery used to examine metabolic and behavioral phenotypes in mice with mutations in specific glycan-binding proteins and glycosyltransferases genes. Because the potential consequences of these genetic deletions are varied, a large number of assays across a variety of domains was included in the battery. The power and usefulness of this approach is in discovering areas for more detailed investigation. PMID- 17132499 TI - Electrophysiological analysis of interactions between carbohydrates and transmitter receptors reconstituted in lipid bilayers. AB - The investigation of functional interactions between carbohydrates and neurotransmitter receptors is a challenging task. The presence of a wide variety of carbohydrates in the nervous system may preclude electrophysiological analysis using intact brain slice preparations or isolated neurons. The purification of transmitter receptors and their subsequent reconstitution into an artificial lipid bilayer can serve as a valuable tool to study carbohydrate and transmitter receptor interaction in a controlled environment. The "tip-dip" bilayer technique along with patch clamp electronics provides a unique means to explore carbohydrate interactions with a single transmitter receptor channel. This technique is also helpful in analyzing the interaction of carbohydrates with synaptic transmitter receptors using isolated synaptosomal preparations. Here, we illustrate the methods involved in reconstituting transmitter receptors in tip dip bilayers and the subsequent study of carbohydrate interaction with the receptors. PMID- 17132500 TI - Methods for analysis of O-linked modifications on epidermal growth factor-like and thrombospondin type 1 repeats. AB - The identification of novel forms of O-linked glycosylation on epidermal growth factor and thrombospondin type 1 repeats, and their emerging functional significance, require the development of new methods for their analysis. This chapter describes detailed methods to analyze both the structure and the site of modification of O-fucose and O-glucose glycans on proteins. These methods use both traditional biochemical methods of carbohydrate composition analysis and electrospray ionization-mass spectrometry of glycopeptides. PMID- 17132501 TI - Roles of O-fucosyltransferase 1 and O-linked fucose in notch receptor function. AB - O-fucosyltransferase 1 (Ofut1) is a soluble endoplasmic reticulum protein that directly transfers fucose onto serine or threonine residues of EGF domain containing proteins such as Notch receptors. Genetic analysis indicates that Ofut1 is essential for Notch receptor activation. To explore the molecular basis for the absolute requirement of Ofut1 for Notch function, biochemical and cell biological approaches were used. Ligand-binding assay revealed that Ofut1 is essential for Notch receptors to physically interact with their ligands. In addition, secretion assay and cell surface staining showed that secretion of Notch receptors is impaired in OFUT1-depleted cells, indicating that the structure of Notch receptors is altered. Interestingly, promotion of Notch secretion by OFUT1 does not require its enzyme activity. Together with the fact that OFUT1 physically associates with Notch and that OFUT1 prevents misfolding of Notch mutants, it is proposed that OFUT1 acts as a chaperone that promotes the folding of the EGF repeat of Notch receptors. This chapter focuses on the methods used to analyze the roles of Ofut1 in Notch receptor structure and function. PMID- 17132502 TI - Roles of O-fucose glycans in notch signaling revealed by mutant mice. AB - Notch receptor signaling is important for many developmental processes in the metazoa. Insights into how Notch receptor signaling is regulated have been obtained from the characterization of mutants of model organisms in which Notch signaling is perturbed. Here we describe the effects of mutations that alter the glycosylation of Notch receptors and Notch ligands in the mouse. The extracellular domain of Notch receptors and Notch ligands carries N-glycans and O glycans, including O-fucose and O-glucose glycans. Mutations in several genes that inhibit the synthesis of O-fucose glycans, and one that also affects the maturation of N-glycans, cause Notch signaling defects and disrupt development. PMID- 17132503 TI - Defect in glycosylation that causes muscular dystrophy. AB - Muscular dystrophies are a diverse group of inherited disorders characterized by progressive muscle weakness and wasting. The dystrophin-glycoprotein complex is composed of alpha-, beta-dystroglycan (DG), dystrophin and some other molecules. alpha- and beta-DG stabilize the sarcolemma by acting as an axis through which the extracellular matrix is tightly linked to the cytoskeleton. The relative molecular weights of alpha-DG differ in different tissues as a result of differential glycosylation. New findings indicate that disrupted glycosylation of alpha-DG results in a loss of ligand binding, giving rise to both progressive muscle degeneration and abnormal neuronal migration in the brain. This article discusses methods, including purification of alpha-DG and glycosyltransferase assays involved in alpha-DG glycosylation. PMID- 17132504 TI - Identification and analysis of novel glycolipids in vertebrate brains by HPLC/mass spectrometry. AB - Glycosphingolipids are a major component of microdomains or lipid rafts in biological membranes. A new member of raft glycolipids, phosphatidylglucoside (PtdGlc), as well as 6-O-Ac-PtdGlc, a form of PtdGlc O-acetylated at position 6 of its glucopyranose ring, is present in central nervous system tissues. Because the glycolipids represent a minor constituent of lipid rafts and because their mass numbers are the same as that of phosphatidylinositol (PI), the glycolipids are difficult to detect and purify. Here we describe methods to purify and identify glycolipids from rodent brain and methods to discriminate PtdGlc from PI in chick spinal cord using HPLC/electrospray ionization ion-trap mass spectrometry. PMID- 17132505 TI - Modulation of growth factor signaling by gangliosides: positive or negative? AB - Increasing evidence has implicated gangliosides, sialic acid-containing cell surface glycosphingolipids, in the biological and clinical behavior of many types of human tumors. Gangliosides are overexpressed and actively shed by tumor cells, can bind to normal cells in the tumor microenvironment, and have a number of biological properties that could conceivably alter tumor-host interactions to influence the survival of the malignant cells that carry these molecules. One major area of investigation is the modulation of cell signaling by gangliosides. Published studies have demonstrated modulation of growth factor signaling through the epidermal growth factor (EGF), fibroblast growth factor (FGF), platelet derived growth factor (PDGF), Trk family, and insulin receptors. Studies conducted over the past 10 y have demonstrated either inhibition or enhancement of signaling by gangliosides, depending on cell type, ganglioside species, and experimental conditions. Of particular concern are conflicting studies that demonstrate opposite effects of gangliosides on the same growth factor receptor. This chapter discusses a methodological approach to addressing this apparent conflict. PMID- 17132506 TI - Activation of natural killer T cells by glycolipids. AB - Natural killer T (NKT) cells are a distinct T-cell sublineage, originally named because of their coexpression of an alphabeta T cell antigen receptor (TCR) characteristic of T lymphocytes, and NK1.1, a C-type lectin expressed by natural killer (NK) cells. NKT cells use their TCR to recognize glycolipids bound to or presented by CD1d. Until recently, most studies used the synthetic glycolipid alpha-galactosylceramide (alphaGalCer) to activate these lymphocytes, and very little was known about the natural antigens recognized by NKT cells. Given the pivotal role played by the NKT cells in many immune responses, including antimicrobial responses, tumor rejection, and the development of autoimmune diseases, the identification of the natural antigens recognized by these cells, and analogs that may alter their cytokine production, are goals of primary importance. This chapter discusses methods that can be used to assess the potency of potential glycolipid antigens for this unique population of T lymphocytes, including methods for in vitro NKT cell activation and expansion, in vivo activation, and measurement of their avidity for different antigens. PMID- 17132507 TI - Determination of glycolipid-protein interaction specificity. AB - Glycolipids are found on all eukaryotic cells. Their expression varies among tissues, with the highest density found in the brain, where glycolipids are the most abundant of all glycoconjugate classes. In addition to playing roles in membrane structure, glycolipids also act as cell surface recognition molecules, mediating cell-cell interactions, as well as binding certain pathogens and toxins. Because of their amphipathic nature, underivatized glycolipids are amenable to immobilization on hydrophobic surfaces, where they can be probed with lectins, antibodies, pathogens, toxins, and intact cells to reveal their binding specificities and affinities. Three particularly useful methods to probe specific glycolipid-mediated recognition events are microwell adsorption (ELISA), thin layer chromatography overlay, and surface plasmon resonance (SPR) spectroscopy. PMID- 17132508 TI - Analysis of carbohydrate-carbohydrate interactions using gold glyconanoparticles and oligosaccharide self-assembling monolayers. AB - Carbohydrates are the most extended structures exposed at the surface of most cells. These carbohydrate chains, when arranged in polyvalent clusters, offer a rich supply of low-affinity binding sites, making them a reliable and flexible system to regulate cell adhesion and recognition. The very first model system for cell-cell recognition by means of carbohydrate-carbohydrate interactions in the animal kingdom came from a primitive invertebrate animal: the marine sponge. During the past 50 years, studies have shown that highly repetitive carbohydrate motives on extracellular proteoglycan supramolecular complexes of marine sponge cells are involved in the species-specific adhesion. In this chapter, some glyconanotechnology procedures are described for the detailed investigation of the role of a carbohydrate epitope in the marine sponge cell recognition. The various protocols are generally applicable in other areas of glycoscience. PMID- 17132509 TI - Galectin interactions with extracellular matrix and effects on cellular function. AB - In vivo, cells exist within a complex mixture of glycoproteins and proteoglycans termed the extracellular matrix (ECM). The components of the ECM are secreted by the cells in that site, and the ECM provides not only a physical support but also outside-in signals that regulate many cellular functions, including cell proliferation, differentiation, migration, and survival. Altering the composition of the ECM can thus significantly alter cell behavior. Many types of cells, including normal and malignant epithelial and mesenchymal cells, secrete galectin 1, a member of the galectin family of lectins, into the ECM surrounding the cells. Galectin-1 is known to regulate many of these same cellular functions (i.e., proliferation, differentiation, migration, and death), so that the presence of galectin-1 in ECM will modify the effects of ECM on cells. In this chapter, we present three types of assays that allow interrogation of the effects of galectin-1 on cell adhesion to ECM, cell migration through ECM, and cell death in ECM, using T-cell lines as a model cell type. PMID- 17132510 TI - Galectins in apoptosis. AB - Galectins are a family of animal lectins with affinity for beta-galactosides. By using recombinant proteins, a number of galectins have been shown to interact with cell-surface and extracellular matrix glycoconjugates through lectin carbohydrate interactions. Through this action, they can affect a variety of cellular processes, and the most extensively documented function is induction of apoptosis. By using gene transfection approaches, galectins have been shown to regulate various cellular processes, including apoptosis. Evidence has been provided that some of these functions involve binding to cytoplasmic and nuclear proteins, through protein-protein interactions, and modulation of intracellular signaling pathways. Thus, some galectins are pro-apoptotic, whereas others are anti-apoptotic; some galectins induce apoptosis by binding to cell surface glycoproteins, whereas others regulate apoptosis through interactions with intracellular proteins. This review describes involvement of galectin-1, -2, -3, 7, -8, -9, and -12 in apoptosis. PMID- 17132511 TI - On the role of galectins in signal transduction. AB - Galectins are a family of proteins that bind to beta-galactoside carbohydrate structure through their carbohydrate recognition domains (CRDs). These proteins have been shown to be involved in multiple biological functions such as cell matrix and cell-cell interactions, cell proliferation, cell differentiation, cellular transformation, or apoptosis mainly through their binding properties to specific ligands. Signal transduction should be dramatically affected and changed in the process of those biological functions; namely, galectins can also be involved in several signal transduction pathways. This chapter discusses the role of galectins in signal transduction, dividing into extracellular and intracellular galectins. In addition, we will indicate the methods to identify the interactions of galectins in signal transduction. PMID- 17132513 TI - Caenorhabditis elegans carbohydrates in bacterial toxin resistance. AB - The major virulence factor produced by the bacterium Bacillus thuringiensis (Bt) is a pore-forming toxin called crystal (Cry) toxin, which targets and kills insects and nematodes. To understand how this bacterial toxin interacts with its invertebrate hosts, a genetic screen in C. elegans for nematodes resistant to Bt toxin was carried out. Four of the five genes that mutated to toxin resistance encode glycosyltransferases. These genes were found to participate in the biosynthesis of C. elegans glycosphingolipids. These glycolipids in turn were shown to directly bind Bt toxin. Thus, resistance to Bt toxin in C. elegans can develop as a result of loss of glycolipid receptors for the toxin. Here we describe the isolation of Bt toxin resistance mutants in C. elegans, isolation of C. elegans glycolipids, and their separation by thin-layer chromatography, overlay assays to demonstrate direct binding of Bt toxin to glycolipids, and the purification of specific C. elegans glycolipid species. PMID- 17132512 TI - Helicobacter pylori adhesion to carbohydrates. AB - Adherence of bacterial pathogens to host tissues contributes to colonization and virulence and typically involves specific interactions between bacterial proteins called adhesins and cognate oligosaccharide (glycan) or protein motifs in the host that are used as receptors. A given pathogen may have multiple adhesins, each specific for a different set of receptors and, potentially, with different roles in infection and disease. This chapter provides strategies for identifying and analyzing host glycan receptors and the bacterial adhesins that exploit them as receptors, with particular reference to adherence of the gastric pathogen Helicobacter pylori. PMID- 17132514 TI - N-glycans are involved in the response of Caenorhabditis elegans to bacterial pathogens. AB - Caenorhabditis elegans is becoming a popular tool for the study of glycan function particularly as it applies to development. More than 150 C. elegans genes have been identified as homologs of vertebrate genes involved in glycan metabolism. However, only a relatively small number of these genes have been expressed and studied in any detail. Oligomannose N-glycans (Man5-9GlcNAc2Asn), major components of the N-glycans of all eukaryotes including C. elegans, are essential, at least in part, for eukaryote survival, because they play an important role in protein quality control. In addition, vertebrates make hybrid (GlcNAcMan3-5GlcNAc2Asn) and complex (XGlcNAc2-6Man3GlcNAc2Asn) but little or no paucimannose (Man3-4GlcNAc2Asn)N-glycans, whereas plants, insects, and C. elegans make paucimannose but little or no hybrid nor complex N-glycans. UDP GlcNAc:alpha3-D-mannoside beta1,2-N-acetylglucosaminyltransferase I (encoded by the gene Mgat1) controls the synthesis of hybrid, complex, and paucimannose N glycans in all eukaryotes. C. elegans has three genes encoding beta1,2-N acetylglucosaminyltransferase I (gly-12, gly-13, gly-14). To determine the functional requirement for this enzyme in worms, we generated seven worm strains with mutations in these three genes (gly-12, dpy-6 gly-13, gly-14, gly-12 gly-13, gly-14;gly-12, gly-14;dpy-6 gly-13 and gly-14;gly-12 gly-13). Whereas mice and Drosophila melanogaster with null mutations in Mgat1 suffer severe developmental abnormalities, all seven C. elegans strains with null mutations in the genes encoding beta1,2-N-acetylglucosaminyltransferase I develop normally and seem to have a wild-type phenotype. We now present evidence that beta1,2-N acetylglucosaminyltransferase I-dependent N-glycans (consisting mainly of paucimannose N-glycans) play a role in the interaction of C. elegans with pathogenic bacteria, suggesting that these N-glycans are components of the worm's innate immune system. PMID- 17132515 TI - Detection of cytoplasmic glycosylation associated with hydroxyproline. AB - A special class of glycosylation occurs on a proline residue of the cytoplasmic/nuclear protein Skp1 in the social amoeba Dictyostelium. For this glycosylation to occur, the proline must first be hydroxylated by the action of a soluble prolyl 4-hydroxylase acting on the protein. Cytoplasmic prolyl 4 hydroxylases are dioxygen-dependent enzymes that have low affinity for their O2 substrate and, therefore, have been implicated in O2-sensing in Dictyostelium, as well as in vertebrates and invertebrates. The sugar-hydroxyproline linkage has low abundance, is resistant to alkali cleavage and known glycosidases, and does not bind known lectins. However, initial screens for this modification can be made by assessing changes in electrophoretic mobility of candidate proteins after treatment of cells with prolyl hydroxylase inhibitors, and/or by metabolic labeling with [3H]sugar precursors. In addition, cytoplasmic hydroxylation/glycosylation can be assessed by assaying for cytoplasmic glycosyltransferases. Here we describe these methods and examples of their use in analyzing Skp1 glycosylation in Dictyostelium and the apicomplexan Toxoplasma gondii, the causative agent of toxoplasmosis in humans. PMID- 17132517 TI - The resuscitation greats. Dr. William Cullen and Lord Cathcart. PMID- 17132519 TI - Purchasing health insurance coverage for smoking cessation treatment: employers describe the most influential information in this decision. AB - Employer provision of insurance coverage for smoking cessation treatment (SCT) remains spotty despite a body of treatment efficacy and cost-effectiveness evidence available to inform and support this health care purchasing decision. This qualitative study examined the information on which this coverage decision is made. In this study, state employers describe the content and sources of the most influential information in their decision to provide insurance coverage for SCT as well as a second health benefit for comparative purposes. We provide insight into the extent to which SCT evidence informs the SCT coverage decision and suggest topics and targets for research dissemination. We interviewed 55 employee benefit staff in 35 states. Responses were compared from states with and without SCT coverage to explore the types of information that may be more effective at promoting coverage. The content and sources of the information employers judged most useful varied notably between states with and without SCT coverage. Compelling evidence of the efficacy of SCT and its cost-effectiveness did not appear to play an influential role in the SCT decision among states without SCT coverage relative to states with SCT coverage. States with SCT coverage relied significantly on benefit consultants and actuaries for the information they described as most influential; in comparison, noncovered states reported service providers, staff, and the Internet as major information sources. To foster employers' provision of SCT coverage, research dissemination efforts should emphasize SCT efficacy and cost-effectiveness information and tailor communication to benefit consultants and actuaries in addition to employers themselves. PMID- 17132520 TI - Clinical laboratory evaluation of potential reduced exposure products for smokers. AB - Smoking-related cancer and other disease account for more than 400,000 U.S. deaths annually. Smoking cessation reduces smoking-related disease rates, but relapse rates are high. Thus, interest in reducing the harm of continued smoking is growing. Potential reduced exposure products (PREPs) are marketed to reduce smokers' exposure to smoke toxicants such as carbon monoxide (CO) and carcinogens and may be harm reduction tools. New PREPs are proliferating, but past experience with "low-yield" cigarettes that failed to reduce smokers' toxicant exposure suggests that comprehensive evaluation is necessary to predict if these new products are likely to alter the harm caused by smoking. The purpose of the study was to develop clinical laboratory methods for PREP evaluation. Smokers (N = 35) completed four, 5-day conditions that differed by product used: Advance, Eclipse, own brand cigarettes, or no cigarettes. Carcinogen (as assessed by one nitrosamine and one polycyclic aromatic hydrocarbon biomarker) and nicotine exposure were assessed via thrice-weekly urine sampling. Withdrawal symptoms were measured daily, and smoking behavior was assessed on the first and last day of each condition. Relative to own brand, Advance reduced exposure to the nitrosamine NNK and CO, and Eclipse reduced exposure to nicotine and the nitrosamine NNK, increased exposure to CO, and resulted in larger, longer, and more frequent puffs. No smoking reduced exposure to the nitrosamine NNK, CO, and nicotine, whereas withdrawal was elevated (all p values <.05). Clinical laboratory evaluation of PREPs for smokers is valuable for measuring users' smoke toxicant exposure, withdrawal, and smoking behavior and should be incorporated into a comprehensive PREP evaluation strategy. PMID- 17132521 TI - Does smoking reduction increase future cessation and decrease disease risk? A qualitative review. AB - This review examines whether reduction in smoking among smokers not currently interested in quitting (a) undermines or promotes future smoking cessation or (b) decreases the risks of developing smoking-related diseases. Systematic computer searches and other methods located 19 studies examining reduction and subsequent cessation and 10 studies examining reduction and disease risk. Because of the heterogeneity of methods and results, a meta-analysis could not be undertaken. None of 19 studies found that reduction undermined future cessation, and 16 found that reduction was associated with greater future cessation, including the two randomized trials of reduction versus nonreduction. The 10 trials of disease risk found conflicting results, and none was an adequate test. We conclude that (a) smoking reduction increases the probability of future cessation and (b) whether smoking reduction decreases the risks of smoking-related diseases has not been adequately tested. PMID- 17132522 TI - An evaluation of four measures of adolescents' exposure to cigarette marketing in stores. AB - This study evaluates four measures of exposure to retail cigarette marketing in relation to adolescent smoking behavior. The measures are (a) shopping frequency in types of stores known to carry more cigarette advertising than other store types, (b) shopping frequency in specific stores that sell cigarettes in the study community, (c) the amount of exposure to cigarette brand impressions in stores where students shopped, and (d) perceived exposure to cigarette advertising. The study combined data from classroom surveys administered to 6th-, 7th-, and 8th-grade students in three California middle schools, and direct store observations quantifying cigarette marketing materials and product placement in stores where students shopped. Logistic regression models were used to examine how each exposure measure related to the odds of ever smoking and susceptibility to smoke, controlling for grade, gender, ethnicity, school performance, unsupervised time, and exposure to household and friend smoking. Frequent exposure to retail cigarette marketing as defined by each of the four measures was independently associated with a significant increase in the odds of ever smoking. All but the measure of exposure to store types was associated with a significant increase in the odds of susceptibility to smoke. Four measures of exposure to retail cigarette marketing may serve equally well to predict adolescent smoking but may vary in cost, complexity, and meaning. Depending on the outcomes of interest, the most useful measure may be a combination of self reported exposure to types of stores that contain cigarette marketing and perceived exposure to such messages. PMID- 17132523 TI - Perspectives on the role of the Society for Research on Nicotine and Tobacco in promoting global tobacco research and reducing tobacco harm. AB - The Society for Research on Nicotine and Tobacco (SRNT) has a critical role to play in advancing tobacco research globally. To date SRNT has sponsored a number of successful initiatives including meetings held by the society's European affiliate, establishment of the Global Network Committee in 1999, and the 2003 and 2005 global tobacco pre-conferences. The most recent SRNT conference was held for the first time outside of North America and attracted over 900 participants from 50 countries. SRNT also has sponsored workshops, provided member scholarships to researchers from low- and middle-income countries, and partnered with the World Health Organization to host www.treatobacco.net. Although these initiatives are impressive, SRNT can further support global tobacco research by (a) working with other organizations to support global tobacco science and policy (this is especially timely given the recent coming into force of the Framework Convention on Tobacco Control), (b) providing more resources including increased funding and access to online journals for those in low- and middle-income countries, (c) increasing the representation of science at world and national tobacco conferences, and (d) expanding the role of the Training Committee to place increased emphasis on mentoring tobacco scientists from low- and middle income countries. Science will be crucial in addressing the global tobacco pandemic, and SRNT is uniquely positioned to further tobacco science and research capacity on a global basis. PMID- 17132524 TI - Complementary treatments for tobacco cessation: a survey. AB - Little information is available regarding the prevalence of use and interest in future use of complementary and alternative medicine (CAM) for tobacco cessation among tobacco users. We conducted a self-administered anonymous survey among 1,175 patients seen at a midwestern outpatient tobacco treatment specialty clinic between November 2003 and July 2005. Patient use of CAM for tobacco cessation, perceived efficacy of these treatments, and interest in future use of CAM were ascertained. Data were summarized using descriptive statistics, and logistic regression models were used to determine the characteristics associated with past CAM use or interest in future use of CAM for tobacco cessation. All of the patients who received the survey completed it. A total of 27% of patients reported previous use of CAM for tobacco cessation. The interventions most commonly used were hypnosis, relaxation, acupuncture, and meditation. CAM treatments most commonly perceived to be efficacious were yoga, relaxation, meditation, and massage therapy. A total of 67% of the patients reported interest in future use of CAM for tobacco cessation. The treatments of greatest interest for use in the future were hypnosis, herbal products, acupuncture, relaxation, and massage therapy. Female gender, previous use of conventional tobacco cessation products, previous use of CAM treatments, and a higher level of education were significantly associated with interest in future CAM use. The high level of interest in CAM among tobacco users underscores the need to conduct further research in this field. PMID- 17132525 TI - Changes in markers of epithelial permeability and inflammation in chronic smokers switching to a nonburning tobacco device (Eclipse). AB - Eclipse, produced by R. J. Reynolds Tobacco Company, is a potential reduced exposure product (PREP) that heats rather than burns tobacco. We hypothesized that switching to Eclipse would result in relative normalization of pulmonary epithelial permeability, airway inflammation, and blood leukocyte activation in current smokers. We assessed 10 healthy smokers (aged 21-50 years, 19+/-8 pack years) at baseline and after 2 and 4 weeks of switching to Eclipse, for symptoms, pulmonary function, airway inflammation, lung clearance of (99m)technicium diethylenetriaminepentaacetic acid, and blood leukocyte activation and production of reactive oxygen species. Values were compared before and after Eclipse use and with those of healthy, lifetime nonsmokers (aged 18-53 years). Compared with baseline values before switching to Eclipse, lung permeability half-time increased from 33+/-3 to 43+/-6 min (p = .017) after 2 weeks and to 44+/-7 min (p = .10) after 4 weeks of Eclipse use. Carboxyhemoglobin levels increased from 5%+/ 2% to 7%+/-2% (p<.01) at 4 weeks. Compared with smoking the usual brand of cigarettes, after smoking Eclipse the percentage of natural killer cells, the expression of intercellular adhesion molecule-1 on monocytes, and the expression of CD45RO on T cells showed significant improvement. However, expression of other surface markers, notably CD23 on monocytes, became more abnormal. Production of reactive oxygen species by smokers' neutrophils and monocytes increased further with Eclipse use. We found no significant effects on pulmonary function, cells in induced sputum, or exhaled nitric oxide. Switching to Eclipse reduces alveolar epithelial injury in some smokers but may increase carboxyhemoglobin levels and oxidative stress. PMID- 17132526 TI - Smoking cessation in general practice: the effects of a quitline. AB - This cross-sectional study assessed changes between 1999 and 2003 in attitudes among Swedish general practitioners (GPs) toward smoking cessation activities and also assessed the effect of a nationwide quitline on GPs' smoking cessation activities. A random sample of 621 Swedish GPs answered a questionnaire mailed to their home addresses in spring 2003. When possible, the results of the present study were compared with results from a similar study conducted in 1999. Main outcome measures were GPs' self-reported activities, perceived barriers to engaging in smoking cessation, and referrals to the quitline. Between 1999 and 2003, GPs increased their overall smoking cessation activities and were more aware of the complexity of smoking cessation support. Significantly more GPs experienced smoking cessation support as "too time consuming" and preferred to refer smokers to counselors specializing in smoking cessation. GPs referring patients to the quitline were more likely to be active in other smoking cessation activities. One out of five GPs had advised their patients to use oral smokeless tobacco as a means to stop smoking. A paradigm shift regarding awareness of the complexity of smoking cessation support may be ongoing amongst Swedish GPs. The nationwide smoking cessation quitline appears to have had a positive effect on GPs' engagement in smoking cessation. PMID- 17132527 TI - Findings and implications from a national study on potential reduced exposure products (PREPs). AB - Tobacco companies have recently introduced products that they claim have reduced toxins and carcinogens, and that they say may be less harmful to smokers. These are potential reduced exposure products, or PREPs. This study measured smokers' awareness of PREPs, use of PREPs, interest in trying PREPs, and beliefs about the regulation of PREPs. This study was based on nationally representative data collected in 2002 and 2003 through the American Smoking and Health Survey. The final sample included 1,174 adult smokers. Descriptive and multivariate analyses were conducted to produce estimates and explore potential correlates of the outcomes. A total of 41.9% of adult smokers reported having heard of at least one of the PREPs measured, and 11.0% reported having tried one of these products. Half of adult smokers (49.9%) said they would like to try PREPs. Interest in trying PREPs was associated with having made a quit attempt, being concerned about the effect of smoking on one's health, and having a household income of less than US dollars 20,000. About half of adult smokers (49.1%) incorrectly believed that PREPs are evaluated for safety by the government before being placed on the market, and 84.2% believed that the government should evaluate the safety of PREPs before they are sold to consumers. This study provides new and timely information on the use of, interest in trying, and beliefs about the regulation of PREPs among a nationally representative sample of adult smokers. With half of adult smokers interested in trying PREPs, the need for concrete scientific evidence on the potential impact of these products is critical. PMID- 17132528 TI - Biomechanics of changes in ACL and PCL material properties or prestrains in flexion under muscle force-implications in ligament reconstruction. AB - The effects of changes in cruciate ligament material and prestrain on knee joint biomechanics following ligament reconstruction surgery by a tendon are not adequately known. A 3D nonlinear finite element model of the entire knee joint was used to investigate the joint response at different flexion angles under a quadriceps force while varying ACL and PCL initial strains or material properties. The ACL and PCL forces as well as tibiofemoral contact forces/areas substantially increased with greater ACL or PCL initial strains or stiffness. The patellofemoral contact force slightly increased whereas the tibial extensor moment slightly decreased with tenser or stiffer ACL. Reverse trends were predicted with slacker ACL. Results confirm the hypotheses that changes in the prestrain of one cruciate ligament substantially influence the force in the other cruciate ligament and the entire joint and that the use of the patellar tendon (PT) as a replacement for cruciate ligaments markedly alters the joint biomechanics with trends similar to those predicted when increasing prestrains. Forces in both ACL and PCL ligaments increased as one of them became tenser or stiffer and diminished as it became slacker. These results have important consequences in joint biomechanics following ligament injuries or replacement and tend to recommend the use of grafts with smaller prestrains (i.e. slacker than intact) when using the PT as the replacement material with stiffness greater than that of replaced ligament itself. PMID- 17132529 TI - Modeling investigation of learning a fast elbow flexion in the horizontal plane- prediction of muscle forces and motor units action. AB - Experimental investigation of practicing a dynamic, goal-directed movement reveals significant changes in kinematics. Modeling can provide insight into the alterations in muscle activity, associated with the kinematic adaptations, and reveal the potential motor unit (MU) firing patterns that underlie those changes. In this paper, a previously developed muscle model and software (Raikova and Aladjov, Journal of Biomechanics, 35, 2002) have been used to investigate changes in MU control, while practicing fast elbow flexion to a target in the horizontal plane. The first trial (before practice) and the last trial (after extensive practice) of two subjects have been simulated. The inputs for the simulation were the calculated external moments at the elbow joint. The external moments were countered by the action of three flexor muscles and two extensor ones. The muscles have been modeled as a mixture of MUs of different types. The software has chosen the MU firing times necessary to accomplish the movement. The muscle forces and MUs firing statistics were then calculated. Three hypotheses were tested and confirmed: (1) peak muscle forces and antagonist co-contraction increase during training; (2) there is an increase in the firing frequency and the synchronization between MUs; and (3) the recruitment of fast-twitch MUs dominates the action. PMID- 17132530 TI - Density-based load estimation using two-dimensional finite element models: a parametric study. AB - A parametric investigation was conducted to determine the effects on the load estimation method of varying: (1) the thickness of back-plates used in the two dimensional finite element models of long bones, (2) the number of columns of nodes in the outer medial and lateral sections of the diaphysis to which the back plate multipoint constraints are applied and (3) the region of bone used in the optimization procedure of the density-based load estimation technique. The study is performed using two-dimensional finite element models of the proximal femora of a chimpanzee, gorilla, lion and grizzly bear. It is shown that the density based load estimation can be made more efficient and accurate by restricting the stimulus optimization region to the metaphysis/epiphysis. In addition, a simple method, based on the variation of diaphyseal cortical thickness, is developed for assigning the thickness to the back-plate. It is also shown that the number of columns of nodes used as multipoint constraints does not have a significant effect on the method. PMID- 17132531 TI - Determination of elastomeric foam parameters for simulations of complex loading. AB - BACKGROUND: Finite element (FE) analysis has shown promise for the evaluation of elastomeric foam personal protection devices. Although appropriate representation of foam materials is necessary in order to obtain realistic simulation results, material definitions used in the literature vary widely and often fail to account for the multi-mode loading experienced by these devices. This study aims to provide a library of elastomeric foam material parameters that can be used in FE simulations of complex loading scenarios. METHOD OF APPROACH: Twelve foam materials used in footwear were tested in uni-axial compression, simple shear and volumetric compression. For each material, parameters for a common compressible hyperelastic material model used in FE analysis were determined using: (a) compression; (b) compression and shear data; and (c) data from all three tests. RESULTS: Material parameters and Drucker stability limits for the best fits are provided with their associated errors. The material model was able to reproduce deformation modes for which data was provided during parameter determination but was unable to predict behavior in other deformation modes. CONCLUSIONS: Simulation results were found to be highly dependent on the extent of the test data used to determine the parameters in the material definition. This finding calls into question the many published results of simulations of complex loading that use foam material parameters obtained from a single mode of testing. The library of foam parameters developed here presents associated errors in three deformation modes that should provide for a more informed selection of material parameters. PMID- 17132532 TI - Application of the finite element method in dental implant research. AB - This article provides a review of the achievements and advancements in dental technology brought about by computer-aided design and the all powerful finite element method (FEM) of analysis. The scope of the review covers dental implants, jawbone surrounding the implant and the biomechanical implant and jawbone interaction. Prevailing assumptions made in the published finite element analysis (FEA) and their limitations are discussed in some detail which helps identify the gaps in research as well as future research direction. PMID- 17132533 TI - Antiendothelial cell antibody levels in healthy normotensives with high normal blood pressure. AB - High normal blood pressure (HNBP) seems to be related to an increased cardiovascular risk in healthy normotensive subjects. According to the literature, elevated levels of antibodies against endothelial cell surface antigen antiendothelial cell antibodies (AECA) play an important role in the early stages of atherosclerosis process and in borderline hypertension. The aim of this study was to compare AECA levels of healthy normotensives with HNBP to those of healthy normotensives with normal blood pressure (NBP), matched for age, sex, and body mass index (BMI). METHODS: Ninety healthy normotensives with HNBP (43M, 47F; mean age, 48 +/- 2.6 yrs; BMI 23.6 +/- 1.5 Kg/m2) (Group A) and 80 healthy normotensives with NBP (41M, 39F; mean age, 46 +/- 3 yrs; BMI 24 +/- 1.8 Kg/m2) (Group B) were studied. Both group subjects were matched for sex, age, and BMI. AECA levels were determined in each subject using an enzyme-linked immunosorbent assay (ELISA). AECA levels were expressed as mean values. RESULTS: Twenty-five subjects from group A (28%) showed elevated IgG antiendothelial cell antibodies levels vs. three from group B (3.75%, p < 0.001). IgM AECA levels were elevated in 18 subjects from group A (20%) vs. two from group B (1.5%, p < 0.001). CONCLUSIONS: The present findings suggest that healthy normotensives with HNBP have significantly higher AECA levels of both classes (IgG, IgM) compared to healthy normotensives with NBP. This may have prognostic significance for the future development of essential hypertension in this group of healthy subjects. PMID- 17132534 TI - Effect of sex hormones on non-esterified fatty acids, intra-abdominal fat accumulation, and hypertension induced by sucrose diet in male rats. AB - Sucrose-fed rats (1) had higher intra-abdominal fat mass and plasma non esterified fatty acids and lower testosterone levels, (2) were hypertensive, and (3) had lower plasma NO metabolites than controls. The lack of testosterone by castration of sucrose-fed rats decreased high blood pressure and circulating non esterified fatty acids and increased NO metabolites. The administration of testosterone to castrated sucrose-fed rats restored hypertension, fat accumulation, and high-circulating non-esterified fatty acids, and lowered NO metabolite levels whereas estradiol treatment did not significantly affect these variables in castrated animals. This study proposes that the low levels of testosterone found in sucrose-fed rats are sufficient to maintain central obesity and increased circulating non-esterified fatty acids, which contribute to the development of hypertension in sucrose-fed rats by modulating the biosynthesis of NO. PMID- 17132535 TI - Awareness and control of hypertension among the elderly in a university hospital. AB - Because medically useful information could also be gathered through hospital based studies, the aim of this study was to assess the prevalence, awareness, treatment, and control of HT among the elderly at a university hospital in Turkey. A total of 3,038 elderly were enrolled. Overall, 75.1% of the patients had HT. Among the hypertensive elderly, 88.9% were aware that they had HT, and only 20.9% had BPs that were under control. The low prevalence of control, even among individuals aware of their HT, suggests that the education of health care providers at all stages of professional training is especially important. PMID- 17132536 TI - Defective renal dopamine D1 receptor function contributes to hyperinsulinemia mediated hypertension. AB - Hyperinsulinemia is reported to play a role in hypertension, as abnormalities in blood pressure regulation and sodium handling exist in diabetes mellitus. Kidney dopamine promotes sodium excretion via the activation of renal D1 receptors. Because there is a close relationship between renal D1 receptor function and sodium excretion, it is hypothesized that a defect in this mechanism may contribute to decreased sodium excretion and hypertension during hyperinsulinemia. Renal D1 receptor function was studied in insulin-induced hypertension in male Sprague Dawley rats. Insulin pellets were implanted subcutaneously for controlled insulin release for three weeks; sham rats served as a control. Compared to control rats, insulin pellets increased plasma insulin levels by eight fold and decreased blood glucose by 40%. Insulin also caused a 22 mmHg increase in mean arterial blood pressure compared to control animals. The intravenous infusion of SKF-38393, a D1 receptor agonist, increased sodium excretion in control rats, but SKF-38393 failed to produce natriuresis in hyperinsulinemic animals. Renal proximal tubules from hyperinsulinemic rats had a reduced D1 receptor number, defective receptor-G protein coupling, and blunted SKF-38393 induced Na, K-ATPase inhibition. Insulin seems to reduce D1 receptor expression and coupling to the G-protein, leading to a reduced D1 receptor mediated Na, K-ATPase inhibition, and a diminished natriuretic response to SKF 38393. These phenomena could account for sodium retention and hypertension associated with hyperinsulinemia. PMID- 17132537 TI - Effect of synthetic corticosteroids on vascular reactivity in the human forearm. AB - Exogenous cortisol raises blood pressure (BP) and suppresses acetylcholine (ACh) induced vasodilatation in healthy male volunteers. This study tests the hypothesis that the activation of either classical type I or II corticosteroid receptors by synthetic corticosteroids induces endothelial dysfunction. In two separate studies, dexamethasone or fludrocortisone was administered to healthy male subjects over five days. BP, metabolic parameters, and forearm blood flow (FBF) responses to intra-arterial ACh and nitroprusside (SNP) were measured on day 5 of treatment. Fludrocortisone (800 microg/day) and dexamethasone (3 mg/day) increased BP from control measurements, but not when compared with placebo. Metabolic effects of the steroids were consistent with their known actions. Endothelium-dependent vasodilatation was enhanced by fludrocortisone, most obviously in the presence of nitric oxide (NO) synthase inhibition with NG-mono methyl-L-arginine (LNMMA). Dexamethasone did not suppress endothelium dependent or independent vasodilatation. Non-NO-mediated endothelium-dependent vasodilatation was increased by systemic mineralocorticoid excess but unaffected by glucocorticoid excess. These results do not support the notion that cortisol induced vascular effects are mediated through classical corticosteroid receptors. PMID- 17132538 TI - Effect of the 2004 Mid-Niigata Prefecture earthquake on home blood pressure measurement in the morning in type 2 diabetic patients. AB - A major earthquake struck the Niigata Prefecture, Japan, on October 23, 2004. This study investigated the effect of the earthquake on morning home blood pressure (MHBP) measurements, as well as clinic blood pressure (CBP) and associated complications, in 222 type 2 diabetic patients who measured MHBP and CBP before the earthquake. Physical and laboratory examinations were assessed at every three months. Each patient completed a questionnaire on MHBP measurement, Japanese intensity grade (JIS), patient's lifestyle and psychological impact using posttraumatic stress disorder (PTSD) symptom score. Median JIS showed all patients lived in areas affected by strong aftershocks. Most patients stayed in their own houses, while one-third of patients sought refuge in other houses. No new clinical manifestations of disease were noted. Median PTSD score was low. Patients in public refuge houses had daytime blood pressure, but MHBP was not measured. In the first month, the number of patients who continued MHBP measurements decreased to 27% of pre-shock level. Many patients were unable to measure MHBP for several reasons, including losing MHBP equipment, having equipment destroyed, or suffering from anxiety due to the extensive devastation. Mean systolic MHBP and median urinary albumin excretion rate (UAER) increased significantly within three months and returned to pre-earthquake level at six months. On multiple regression analysis, increased systolic and diastolic MHBPs were significantly associated with UAER elevation. In type 2 diabetic patients following an earthquake, it is important to develop a device of MHBP measurement for maintaining control of MHBP to prevent vascular complications. PMID- 17132539 TI - Mechanisms underlying regulation of the expression and activities of the mammalian pyruvate dehydrogenase kinases. AB - The mechanisms that control mammalian pyruvate dehydrogenase complex (PDC) activity include its phosphorylation (inactivation) by a family of pyruvate dehydrogenase kinases (PDKs 1 - 4). Here we review new developments in the regulation of the activities and expression of the PDKs, in particular PDK2 and PDK4, in relation to glucose and lipid homeostasis. This review describes recent advances relating to the acute and long-term modes of regulation of the PDKs, with particular emphasis on the regulatory roles of nuclear receptors including peroxisome proliferator-activated receptor (PPAR) alpha and Liver X receptor (LXR), PPAR gamma coactivator alpha (PGC-1alpha) and insulin, and the impact of changes in PDK activity and expression in glucose and lipid homeostasis. Since PDK4 may assist in lipid clearance when there is an imbalance between lipid delivery and oxidation, it may represent an attractive target for interventions aimed at rectifying abnormal lipid as well as glucose homeostasis in disease states. PMID- 17132540 TI - Insulin-status-dependent modulation of FoF1 ATPase activity in rat kidney mitochondria. AB - The early and late effects of alloxan-diabetes and insulin treatment on kinetic properties of mitochondrial FoF1 ATPase were examined. Diabetic state resulted in significant decrease in the activity while insulin treatment caused hyper stimulation. In control animals the enzyme activity resolved in three kinetic components. In diabetic condition only component I and II were present. With insulin treatment component III was restored but component II was abolished. Diabetic state and insulin treatment had varied effects on Km values of the three components, whereas the Vmax values were generally on the higher side. Evaluation of the AppKcat/Km values revealed that diabetic state resulted in increased catalytic efficiency; insulin treatment brought back these values to normality. Temperature kinetics studies indicated that the phase transition temperature decreased significantly in the diabetic and insulin-treated diabetic animals. The energy of activation in low temperature range increased in the diabetic animals. Insulin treatment corrected the Arrhenius pattern at early stage of diabetes; at late stage the pattern was reversed. The results are suggestive of subtle insulin status-dependent alterations in membrane structure - function relationships. PMID- 17132541 TI - Development of impaired glucose tolerance and diabetes in follow-up offspring of Caribbean patients with type 2 diabetes: analysis of 5-year follow-up study. AB - Several reports agreed that the antecedent markers for developing diabetes in offspring of type 2 diabetic patients involve excess body weight and insulin resistance. This study examined the pattern of changes in anthropometric and biochemical risk factors for developing diabetes in a follow-up offspring of Caribbean type 2 diabetic patients. Results of 46 offspring of type 2 diabetic patients who had received one-to-one individualized diet and exercise counseling for 5 years in our laboratory were analyzed. Changes in anthropometric (body weight, waist circumference) and biochemical (insulin, glucose, lipids, HOMA insulin resistance, HOMA-percent beta-cell function) parameters over the 5-year period were analyzed using ANOVA tests. Of the 46 offspring, 10.9 and 2.2%, respectively, developed impaired glucose tolerance (IGT) and diabetes. Over the years, IGT offspring had a significant step-wise increase and decrease in fasting and 2-h postprandial plasma glucose levels (P < 0.05) and percent B-cell function (P < 0.001), respectively. Again, a non-significant step-wise increase was observed in body mass index, waist circumference and HOMA-insulin resistance levels (P > 0.05). While we await the results of medication-based intervention studies in different populations, exercise and diet counseling will remain the only available lifestyle intervention strategy for slowing IGT progression to diabetes. PMID- 17132542 TI - Identification of a second-site suppressor mutation of the GTPase defect associated with McCune-Albright syndrome: a model using the yeast heterotrimeric G protein, GPA1. AB - McCune-Albright syndrome (MAS) causes a variety of bone and endocrine abnormalities due to the post-zygotic mutation of the alpha subunit of the stimulatory G-protein Gsalpha. This mutation causes signal-independent activity of the G-protein in the affected cells. We report the development of a system to study the effects of MAS mutations using Saccharomyces cerevisiae, wherein activation of the yeast G-protein pathway results in growth arrest in a genetically recessive fashion. We introduced the MAS mutation into the analogous site in the yeast Galpha gene, GPA1 and randomly mutated the gene to produce intragenic suppressors. Yeast with normal and mutated G-protein genes were induced to lose the normal gene, and mutations able to intragenically suppress the constitutive activity of the MAS mutation were identified based on their ability to form colonies. We report one mutation in GPA1, also in the active site, that is an intragenic suppressor of the MAS defect. PMID- 17132543 TI - Effect of N-benzoyl-D-phenylalanine and metformin on insulin receptors in neonatal streptozotocin-induced diabetic rats: studies on insulin binding to erythrocytes. AB - In the present study, we focused on the insulin-receptor binding in circulating erythrocytes of N-benzoyl-D-phenylalanine (NBDP) and metformin in neonatal streptozotocin (nSTZ)-induced male Wistar rats. We measured blood levels of glucose and plasma insulin and the binding of insulin to cell-membrane ER receptors in NBDP and metformin-treated diabetic rats. The mean specific binding of insulin to ER was significantly lower in diabetic control rats (DC) (53.0 +/- 3.1%) than in NBDP (62.0 +/- 3.1%), metformin (66.0 +/- 3.3%) and NBDP and metformin combination-treated (72.0 +/- 4.2%) diabetic rats, resulting in a significant decrease in plasma insulin. Scatchard plot analysis demonstrated that the decrease in insulin binding was accounted for by a lower number of insulin receptor sites per cell in DC rats when compared with NBDP and metformin-treated rats. High-affinity (Kd1), low-affinity (Kd2), and kinetic analysis revealed an increase in the average receptor affinity in ER from NBDP and metformin-treated diabetic rats having NBDP 2.0 +/- 0.10 x 10(-10) M(-1) (Kd1); 12.0 +/- 0.85 x 10( 8) M(-1) (Kd2), Metformin 2.1 +/- 0.15 x 10(-10) M(-1) (Kd1); 15.0 +/- 0.80 x 10( 8) M(-1) (Kd2), NBDP and metformin 2.7 +/- 0.10 x 10(-10) M(-1) (Kd1); 20.0 +/- 1.2 x 10(-8) M(-1) (Kd2) compared with 0.9 +/- 0.06 x 10(-10) M(-1) (Kd1); 6.0 +/ 0.30 x 10(-8) M(-1) (Kd2) in DC rats. The results suggest an acute alteration in the number of insulin receptors on ER membranes in nSTZ induced diabetic control rats. Treatment with NBDP along with metformin significantly improved specific insulin binding, with receptor number and affinity binding reaching almost normal non-diabetic levels. The data presented here show that NBDP along with metformin increase total ER membrane insulin binding sites with a concomitant significant increase in plasma insulin. PMID- 17132544 TI - A pathophysiological approach to metabolic syndrome using factor analysis in an adult Romanian population. AB - The aim of the study was to examine the role of insulin resistance in etiopathogenesis of metabolic syndrome in an adult Romanian population using exploratory factor analysis. We analyzed 228 non-diabetic subjects randomized in respect to the age and sex distribution of the general population. For each patient, age, sex, body mass index (BMI), systolic and diastolic blood pressure (SBP, DBP), HDL-cholesterol (HDL), plasma triglycerides (TG), fasting plasma glucose (FPG) and fasting insulin were obtained. Factor analysis was performed using principal component analysis, with Varimax rotation of the major determinants of metabolic syndrome. Mean age was 48.9 +/- 12.7 years; 107 (46.9%) were men and 121 (53.1%) women. We found three major factors, which are correlated with metabolic syndrome and may explain its variance. Factor 1 comprises SBP and DBP in men and SBP, DBP and BMI in women. Factor 2 comprises BMI, HDL, TG and FPG in men and BMI, TG and FPG in women. Factor 3 comprises fasting insulin in men and fasting insulin, TG and HDL in women. The finding of more than one factor suggests that insulin resistance is not the only pathophysiological mechanism involved. These factors appear to work independently of each other in men, but they intersect in women, suggesting that the pathophysiology of metabolic syndrome may be different in women compared with men. PMID- 17132545 TI - Factors triggering relatives' judgements of personality change after traumatic brain injury. AB - OBJECTIVE: This study assesses neuropsychological and psychosocial determinants of judgements of personality change (PC) after traumatic brain injury (TBI). DESIGN: Cross-sectional data from 87 participants with TBI analysed using hierarchical binary regression. METHODS: Interview of participants assessed, injury severity (post-traumatic amnesia), degree of cognitive impairment (IQ and memory), presence of orbitofrontal and/or medial temporal damage (olfaction) and emotional reactions (anger, depression and anxiety questionnaires). Separate interview of relatives assessed their own response to the injury (emotional reactions and critical comments from the Expressed Emotion index) and their judgements of participants' behavioural change. RESULTS: As expected, memory loss and impaired olfaction predicted relatives' PC judgements. However, the most striking and novel result was that social-emotional factors (participants' and relatives' reactions) predicted PC judgements best. CONCLUSIONS: This study emphasizes the need (a) for future studies measuring social behaviours (e.g. impulsivity and emotional perception) (b) to explore the differing determinants of relatives' and participants' PC judgements and (c) to develop and to tailor clinical responses to the various psychosocial determinants of PC. PMID- 17132546 TI - The factor structure of the Hospital Anxiety and Depression Scale in acquired brain injury. AB - PRIMARY OBJECTIVE: To evaluate the factor structure of the Hospital Anxiety and Depression Scale (HADS) as a mood assessment in a sample of individuals with acquired brain injury (ABI). RESEARCH DESIGN: Cohort study. METHODS AND PROCEDURES: HADS data from 140 people with ABI was subjected to principle components analysis (PCA). EXPERIMENTAL INTERVENTIONS: None. MAIN OUTCOMES AND RESULTS: A three-factor solution emerged. Items loading on the first two factors map on to the anxiety and depression scales. However, two items from the depression sub-scale did not load significantly on either of these two main factors. Possible reasons are discussed. CONCLUSIONS: This study suggests that the HADS is a useful tool for examining depression and anxiety in a brain injured population, but interpretation of responses to some items should be cautious. PMID- 17132547 TI - Recovery of components of memory in post-traumatic amnesia. AB - PRIMARY OBJECTIVE: Post-traumatic amnesia by definition indicates significant impairment of new learning ability, however very few studies have, examined the natural history and resolution of memory and new learning during PTA. Those studies which have, tended to examine orientation separately from the memory processes required to achieve orientation. Analysis of the order of recovery of the items of the Westmead PTA scale was used to examine recovery of memory and new learning capacity. METHODS: The results of daily assessment of 34 patients with traumatic brain injury (TBI) on the Westmead PTA scale were analysed for order of recovery. RESULTS: The pattern of rank order of item recovery indicated that Date of Birth recovered consistently first. There was variability in the remaining items, however items reflecting long-term memory tended to recover second and items reflecting simple new learning followed. Recall of all three pictures reflecting complex new learning recovered last. CONCLUSION: The pattern of recovery of memory and new learning during PTA reflects a number of complex, inter-related variables including; the familiarity with the information, amount of rehearsal both before and since the accident and the number of cues available in the environment. PMID- 17132548 TI - The rapid screen of concussion: an evaluation of the non-word repetition test for use in mTBI research. AB - PRIMARY OBJECTIVES: (1) To investigate the Nonword Repetition test (NWR) as an index of sub-vocal rehearsal deficits after mild traumatic brain injury (mTBI); (2) to assess the reliability, validity and sensitivity of the NWR; and (3) to compare the NWR to more sensitive tests of verbal memory. RESEARCH DESIGN: An independent groups design. METHODS AND PROCEDURES: Study 1 administered the NWR to 46 mTBI and 61 uninjured controls with the Rapid Screen of Concussion (RSC). Study 2 compared mTBI, orthopaedic and uninjured participants on the NWR and the Hopkins Verbal Learning Test (HVLT-R). MAIN OUTCOMES AND RESULTS: The NWR did not improve the diagnostic accuracy of the RSC. However, it is reliable and indexes sub-vocal rehearsal speed. These findings provide evidence that although the current form of the NWR lacks sensitivity to the impact of mTBI, the development of a more sensitive test of sub-vocal rehearsal deficits following mTBI is warranted. PMID- 17132549 TI - Assessment of intra-cranial pressure after severe traumatic brain injury by transcranial Doppler ultrasonography. AB - PRIMARY OBJECTIVE: To investigate the potential of transcranial Doppler ultrasonography in estimating post-traumatic intra-cranial pressure early after severe traumatic brain injury. RESEARCH DESIGN: The group of 24 patients was analysed for the observation of an early post-traumatic cerebral haemodynamic by middle cerebral artery blood velocity measuring. METHODS AND PROCEDURES: The standard method of measuring the mean blood middle cerebral artery velocity by transcranial Doppler ultrasonic device was performed. MAIN OUTCOMES AND RESULTS: The increased duration of intra-cranial hypertension correlated to the middle cerebral artery low blood velocity (p = 0.042; r = -0.498) (n = 17) and to elevated pulsatility indices (p = 0.007; r = 0.753) (n = 11) significantly. The increased duration of lowered cerebral perfusion pressure correlated to the middle cerebral artery low blood velocity significantly (p = 0.001; r = -0.619) (n = 24). CONCLUSIONS: The significance of transcranial Doppler ultrasonography as a method to estimate an early post-traumatic intra-cranial pressure after severe brain injury was confirmed. This simple and non-invasive technique could be easily used in daily clinical practice and precede intra-cranial pressure monitoring in selected patients. PMID- 17132550 TI - Subjective outcome of brain injury rehabilitation in relation to the therapeutic working alliance, client compliance and awareness. AB - OBJECTIVE: To investigate the relationship between working alliance, compliance, awareness and subjective outcome of brain injury rehabilitation. Subjects were 86 patients who were clients in an holistic neuropsychological outpatient rehabilitation programme. They had suffered a traumatic brain injury (n = 27), a cerebrovascular accident (n = 49) or some other neurological insult (n = 10). MEASURES: The therapeutic alliance, clients' awareness and their compliance were rated four times during the 14-week rehabilitation programme. The therapeutic alliance was rated by both clients and therapist using the Working Alliance Inventory (WAI), awareness and compliance were rated by the therapists. Clients completed the European Brain Injury Questionnaire (EBIQ) at programme start and end. Clients and therapists rated the overall success of their collaboration at programme end. RESULTS: Clients' experience of a good emotional bond between themselves and therapists in mid-therapy was predictive for the reduction of clients' report of depressive symptoms on the EBIQ depression sub-scale (R = 0.68, n = 43, p < 0.001). Good compliance early in the programme was predictive of changes on the EBIQ. Improvement of awareness was related to the amplification of depressive symptoms (r = -0.27, n = 56, p < 0.05). CONCLUSIONS: Brain injury rehabilitation should be seen as a dynamic process that develops between clients and therapists. Future research should further investigate the relationship between process and outcome and how the therapeutic process can be optimized. PMID- 17132551 TI - Email-facilitated qualitative interviews with traumatic brain injury survivors: a new and accessible method. AB - PRIMARY OBJECTIVE: To trial the method of email-facilitated qualitative interviewing with people with traumatic brain injury (TBI). RESEARCH DESIGN: Qualitative semi-structured email-facilitated interviews. PROCEDURES: Nineteen people (17 severe diagnosis) with a TBI participated in email interviews. MAIN OUTCOMES AND RESULTS: Findings indicate that this method facilitates the participation of people with TBI in qualitative interviews. Advantages include increased time for reflection, composing answers and greater control of the interview setting. In addition, the data indicates that people with a TBI are capable of greater insight, reflection and humour than indicated by previous research. CONCLUSION: Findings indicate that new technologies may advance data collection methods for people with cognitive-linguistic impairments who face participation barriers in face-to-face interviews. PMID- 17132552 TI - The influence of neuropsychological rehabilitation on symptomatology and quality of life following brain injury: a controlled long-term follow-up. AB - PRIMARY OBJECTIVE: To establish whether, following acquired brain injury, intensive post-acute neuropsychological rehabilitation could have long-term beneficial effects. METHODS AND PROCEDURES: A group of 37 adults who had suffered cerebrovascular accidents or traumatic brain injuries and who had undergone a rehabilitation programme were followed up 12-22 years post-injury, together with a non-rehabilitated control group of 13 adults, matched for brain-injury and demographics characteristics. Both groups completed a set of questionnaires concerning broad aspects of psychological well-being. Significant others completed similar questionnaires. MAIN OUTCOMES AND RESULTS: The rehabilitation group showed significantly lower levels of brain injury symptoms and higher levels of competency at follow-up. They also rated internal locus of control and general self-efficacy as significantly higher than the control group. Anxiety and depression levels were significantly lower and quality of life significantly higher in the rehabilitation group for both the subjects themselves and for their significant others. CONCLUSIONS: Within methodological limitations this study suggests that post-acute neuropsychological rehabilitation can have long-term beneficial effects. PMID- 17132553 TI - Treating dysarthria following traumatic brain injury: investigating the benefits of commencing treatment during post-traumatic amnesia in two participants. AB - PRIMARY OBJECTIVE: The aims of this preliminary study were to explore the suitability for and benefits of commencing dysarthria treatment for people with traumatic brain injury (TBI) while in post-traumatic amnesia (PTA). It was hypothesized that behaviours in PTA don't preclude participation and dysarthria characteristics would improve post-treatment. RESEARCH DESIGN: A series of comprehensive case analyses. METHODS AND PROCEDURES: Two participants with severe TBI received dysarthria treatment focused on motor speech deficits until emergence from PTA. A checklist of neurobehavioural sequelae of TBI was rated during therapy and perceptual and motor speech assessments were administered before and after therapy. MAIN OUTCOMES AND RESULTS: Results revealed that certain behaviours affected the quality of therapy but didn't preclude the provision of therapy. Treatment resulted in physiological improvements in some speech sub-systems for both participants, with varying functional speech outcomes. CONCLUSIONS: These findings suggest that dysarthria treatment can begin and provide short-term benefits to speech production during the late stages of PTA post-TBI. PMID- 17132556 TI - Preliminary evidence of asymmetry in uncomfortable loudness levels after unilateral hearing aid experience: evidence of functional plasticity in the adult auditory system. AB - The aim of the study was to compare uncomfortable loudness levels (ULLs) in a group of adults before and after unilateral hearing aid experience. Twelve participants presented with a symmetrical hearing loss consistent with natural ageing. Pure tones were presented to each ear separately, commencing at 60 dB HL and increased in 5-dB step sizes until ULL was reached. The post-fitting ULLs were typically measured three years after fitting. Hearing thresholds were symmetrical and remained unchanged after fitting. Mean ULL values were symmetrical before fitting. The mean ULL values increased (i.e. greater tolerance) in both ears after fitting; however, the increase was greatest in the fitted ear: 14.5 and 7 dB at 2000-4000 Hz in the fitted and not-fitted ear, respectively. A separate two-factor repeated ANOVA (ear and frequency) was performed on the pre and post-fitting ULL data. There was no statistically significant difference for ear (p > 0.05) when comparing the pre-fitting ULLs. However, there was a statistically significant difference for ear (p < 0.01) when comparing post-fitting ULLs. The underlying mechanism for the asymmetry is unknown but it is consistent with learning induced reorganization within the auditory system. PMID- 17132557 TI - The effect of intravenously administered mexiletine on tinnitus - a pilot study. AB - The effect of intravenously administered mexiletine on subjective tinnitus and hearing was studied in six patients, who initially responded positively to lidocaine. Distinct mexiletine-induced decreases in tinnitus loudness were demonstrated in three subjects, as reflected by maximum VAS (visual analogue scale) level reduction of 34%, 95%, and 100%, respectively. One subject reported change in tinnitus pitch, another one showed a slight (18% on VAS) tinnitus reduction, and one subject disclosed no effect. Side effects were seen only during one of seven infusions. Mexiletine induced shifts in pure-tone threshold, transient evoked otoacoustic emission, and acoustic reflex threshold, probably reflecting a reversible interference in the function of organ of Corti. The concentration effect relationship remained unclear and no general 'therapeutic' level could be identified. This study confirms the effect of mexiletine on the auditory function and its potential as a possible therapeutic agent or a model for further development in tinnitus pharmacotherapy. PMID- 17132558 TI - Providing an internet-based audiological counselling programme to new hearing aid users: a qualitative study. AB - People with an acquired hearing loss often have difficulty adjusting to a first hearing aid. Studies have shown that audiological counselling can facilitate adjustment to a first hearing aid. Because of its interactive nature, the internet could be a valuable tool to gain information about the experiences of the new hearing aid user and to address the needs for audiological counselling. An internet-based audiological counselling programme in the form of daily e-mails during the first month after the hearing aid fitting was offered to three new hearing aid users. The data, qualitative in nature, were comprised of the content of the e-mails and of in-depth interviews with the participants and their audiologist, and were analysed according to grounded theory. Overall, the internet-based audiological counselling programme provided rich descriptions of the experiences of the participants and reinforced positive adjustment behaviours experienced by them. PMID- 17132559 TI - The effect of a hearing aid noise reduction algorithm on the acquisition of novel speech contrasts. AB - Audiologists are reluctant to prescribe digital hearing aids with active digital noise reduction (DNR) to pre-verbal children due to their potential for an adverse effect on the acquisition of language. The present study investigated the relation between DNR and language acquisition by modeling pre-verbal language acquisition using adult listeners presented with a non-native speech contrast. Two groups of normal-hearing, monolingual Anglophone subjects were trained over four testing sessions to discriminate novel, difficult to discriminate, non native Hindi speech contrasts in continuous noise, where one group listened to both speech items and noise processed with DNR, and where the other group listened to unprocessed speech in noise. Results did not reveal a significant difference in performance between groups across testing sessions. A significant learning effect was noted for both groups between the first and second testing sessions only. Overall, DNR does not appear to enhance or impair the acquisition of novel speech contrasts by adult listeners. PMID- 17132560 TI - Acquired profound hearing loss: mental health and other characteristics of a large sample. AB - The study investigated the mental health and other characteristics of people with acquired profound hearing loss (APHL) and contrasted this group with acquired hearing loss (AHL) in general. A survey was completed over the internet by 95 adults and by 27 people who had attended a one-week course of rehabilitation. The latter group completed questionnaires of anxiety and depression, post-traumatic stress, and hearing handicap. The survey covered a wide range of factors associated with the history of hearing loss and evaluated previous contacts with professional services. The data from the two samples were very similar and were combined. The results indicated the existence of sudden and progressive onset groups, reliance on lip-reading, a severe effect of tinnitus, and some support for the conclusion that the psychosocial impact was greater in APHL than in AHL. A subgroup of APHL was severely distressed and handicapped. Respondents valued medical and audiological services but there was little evidence that previous counselling and support had been helpful. Recommendations for rehabilitation are briefly discussed. PMID- 17132561 TI - Attendance at religious services and physical functioning after stroke among older Mexican Americans. AB - The purpose of this study was to examine the association of attendance at religious services and change in physical functioning among older Mexican Americans who report residual physical limitations due to stroke. Using data from the Hispanic Established Population for the Epidemiological Study of the Elderly (H-EPESE), generalized linear models were used to evaluate change in physical function over 3 years in persons with stroke aged 65 and older, controlling for demographics, medical conditions, health behaviors, and physical mobility. The results showed frequent attendees at religious services had significantly fewer declines in activities of daily living (ADLs) disability compared to infrequent attendees. The frequent attendance group also showed less decline in lower body function compared to the infrequent attendees. Findings are indicative that church attendance prestroke is associated with better physical function poststroke in older Mexican Americans. PMID- 17132562 TI - Genetic and environmental influences on intraindividual variability in reaction time. AB - Reaction time (RT) data from the Minnesota Twin Study of Adult Development and Aging were used to investigate genetic and environmental influences on intraindividual variability in RT. The sample was 738 individuals (aged 27 to 95 years), including 185 monozygotic (MZ) and 131 dizygotic (DZ) twin pairs. Two RT factors were created: decision time and movement time. Age-corrected heritability estimates were 40% for mean RT; 21% for intraindividual variability in movement time; and 0% for intraindividual variability in decision time. Multivariate analysis indicated that all genetic variance in the RT measures was shared among measures; environmental variance was specific to each RT measure. PMID- 17132563 TI - Estimated age effects in athletic events and chess. AB - Rates of decline are estimated using record bests by age for chess and for various track and field, road running, and swimming events. Using a fairly flexible functional form, the estimates show linear percent decline between age 35 and about age 70 and then quadratic decline after that. Chess shows much less decline than the physical activities. Rates of decline are generally larger for the longer distances, and for swimming they are larger for women than for men. An advantage of using best-performance records to estimate rates of decline is that the records are generally based on very large samples. In addition, the age range is large. In this study the age range is 35 to 100 for swimming, 35 to 98 for track and field and running, and 35 to 94 for chess. The estimates also do not suffer from traditional forms of selection bias. PMID- 17132564 TI - Parkinsonian signs and functional disability in old age. AB - Little is known about the association between parkinsonian signs and functional disability. The authors examined the associations between four parkinsonian signs (gait, rigidity, bradykinesia, tremor) and the ability to perform physical and instrumental activities of daily living, in a large cohort of older Rush Memory and Aging Project participants who were free of dementia, Parkinson's disease, and antipsychotic drug use. In a series of logistic regression equations that controlled for age, sex, education, depressive symptoms, cognitive ability, chronic medical conditions, and psychotropic drug use, higher levels of parkinsonian signs were associated with higher levels of disability in performing both physical and instrumental activities of daily living. PMID- 17132565 TI - Correlates of life space in a volunteer cohort of older adults. AB - The authors measured the spatial extent of movement of older persons (i.e., life space) and examined factors that are related to life space. A larger life space was positively correlated with self-report measures of disability. In generalized logit models adjusted for demographics and time of year, a larger life space was associated with less visual impairment, higher levels of lower extremity motor performance, global cognition, and social involvement, and with personality and purpose in life. The results suggest that the range of environmental movement in older adults is a useful indicator of health in old age and may complement measures of disability. PMID- 17132566 TI - Age differences in the bases for social judgments: tests of a social expertise perspective. AB - Research indicates that increasing age is associated with greater use of trait diagnostic behavioral information in making social judgments. These effects may reflect an aging-related increase in social expertise, indicative of more powerful and accessible knowledge structures. The current work is an attempt to provide further evidence in support of the social expertise view and a test of an alternative hypothesis. Results of this work indicate that age differences in the use of trait-diagnostic information were moderated by factors thought to affect the accessibility of relevant knowledge structures. PMID- 17132568 TI - Drug and alcohol policy at the global level. PMID- 17132569 TI - Global use of alcohol, drugs and tobacco. AB - Humans have always used drugs, probably as part of their evolutionary and nutritional heritage. However, this previous biological adaptation is unlikely to be so in the modern world, in which 2 billion adults (48% of the adult population) are current users of alcohol, 1.1 billion adults (29% of the adult population) are current smokers of cigarettes and 185 million adults (4.5% of the adult population) are current users of illicit drugs. The use of drugs is determined largely by market forces, with increases in affordability and availability increasing use. People with socio-economic deprivation, however measured, are at increased risk of harmful drug use, as are those with a disadvantaged family environment, and those who live in a community with higher levels of substance use. Substance use is on the increase in low-income countries which, in the coming decades, will bear a disproportionate burden of substance related disability and premature death. PMID- 17132570 TI - Global burden of disease from alcohol, illicit drugs and tobacco. AB - The use of alcohol, tobacco and illicit drugs entails considerable burden of disease: in 2000, about 4% of the global burden as measured in disability adjusted life years was attributable to each alcohol and tobacco, and 0.8% to illicit drugs. The burden of alcohol in the above statistic was calculated as net burden, i.e. incorporating the protective health effects. Tobacco use was found to be the most important of 25 risk factors for developed countries in the comparative risk assessment underlying the data. It had the highest mortality risk of all the substance use categories, especially for the elderly. Alcohol use was also important in developed countries, but constituted the most important of all risk factors in emerging economies. Alcohol use affected younger people than tobacco, both in terms of disability and mortality. The burden of disease attributable to the use of legal substances clearly outweighed the use of illegal drugs. A large part of the substance-attributable burden would be avoidable if known effective interventions were implemented. PMID- 17132571 TI - Mental health and alcohol, drugs and tobacco: a review of the comorbidity between mental disorders and the use of alcohol, tobacco and illicit drugs. AB - This paper reviews some major epidemiological studies undertaken in high-income countries during the last 15 years which have reported the prevalence of mental disorders and substance use disorders and their relationship. Comorbidity between mental and substance use disorders is highly prevalent across countries. In general, people with a substance use disorder had higher comorbid rates of mental disorders than vice versa, and people with illicit drug disorders had the highest rates of comorbid mental disorders. There is a strong direct association between the magnitude of comorbidity and the severity of substance use disorders. While causal pathways differ across substances and disorders, there is evidence that alcohol is a casual factor for depression, in some European countries up to 10% of male depression. Policies that reduce the use of substances are likely to reduce the prevalence of mental disorders. Treatment should be available in an integrated fashion for both mental and substance use disorders. There is a need to expand the evidence base on comorbidity, particularly in low-income countries. PMID- 17132572 TI - The global economic burden of alcohol: a review and some suggestions. AB - Economic arguments for acting for health are increasingly important for policymakers, yet to date there has been no consideration of the likely economic burden of alcohol on the global level. A review of existing cost estimates was conducted, with each study disaggregated into different cost areas and the methodology of each element evaluated. The range of figures produced from more robust studies was then applied tentatively on the global level. The reviewed studies suggested a range of estimates of 1.3-3.3% of total health costs, 6.4 14.4% of total public order and safety costs, 0.3 - 1.4 per thousand of GDP for criminal damage costs, 1.0-1.7 per thousand of GDP for drink-driving costs, and 2.7-10.9 per thousand of GDP for work-place costs (absenteeism, unemployment and premature mortality). On a global level, this suggests costs in the range of US dollars 210-665 billion in 2002. These figures cannot be understood without considering simultaneously six key problems: (i) the methods used by each study; (ii) who pays these costs; (iii) the 'economic benefits' of premature deaths; (iv) establishing causality; (v) omitted costs; and (vi) the applicability of developed country estimates to developing countries. Alcohol exerts a considerable economic burden worldwide, although the exact level of this burden is a matter of debate and further research. Policymakers should consider economic issues alongside evidence of the cost-effectiveness of particular policy options in improving health, such as in the WHO's CHOICE project. PMID- 17132573 TI - Comparative cost-effectiveness of policy instruments for reducing the global burden of alcohol, tobacco and illicit drug use. AB - Alcohol, tobacco and illicit drug use together pose a formidable challenge to international public health. Building on earlier estimates of the demonstrated burden of alcohol, tobacco and illicit drug use at the global level, this review aims to consider the comparative cost-effectiveness of evidence-based interventions for reducing the global burden of disease from these three risk factors. Although the number of published cost-effectiveness studies in the addictions field is now extensive (reviewed briefly here) there are a series of practical problems in using them for sector-wide decision making, including methodological heterogeneity, differences in analytical reference point and the specificity of findings to a particular context. In response to these limitations, a more generalised form of cost-effectiveness analysis (CEA) is proposed, which enables like-with-like comparisons of the relative efficiency of preventive or individual-based strategies to be made, not only within but also across diseases or their risk factors. The application of generalised CEA to a range of personal and non-personal interventions for reducing the burden of addictive substances is described. While such a development avoids many of the obstacles that have plagued earlier attempts and in so doing opens up new opportunities to address important policy questions, there remain a number of caveats to population-level analysis of this kind, particularly when conducted at the global level. These issues are the subject of the final section of this review. PMID- 17132574 TI - International trade agreements challenge tobacco and alcohol control policies. AB - This report reviews aspects of trade agreements that challenge tobacco and alcohol control policies. Trade agreements reduce barriers, increase competition, lower prices and promote consumption. Conversely, tobacco and alcohol control measures seek to reduce access and consumption, raise prices and restrict advertising and promotion in order to reduce health and social problems. However, under current and pending international agreements, negotiated by trade experts without public health input, governments and corporations may challenge these protections as constraints on trade. Advocates must recognise the inherent conflicts between free trade and public health and work to exclude alcohol and tobacco from trade agreements. The Framework Convention on Tobacco Control has potential to protect tobacco policies and serve as a model for alcohol control. PMID- 17132575 TI - International control of alcohol: alternative paths forward. AB - Alcohol was the first psychoactive substance to be subject to international control, but these agreements between colonial powers have long since fallen away. In the wake of the entry into force of the Framework Convention on Tobacco Control (FCTC), paths forward for international control of alcohol are considered. The choices embedded in the FCTC are discussed, and the justifications for a parallel convention for alcohol considered. An alternative would be scheduling alcohol under one or more of the international drug conventions, most probably the 1971 convention, although the convention would have to be amended to allow use outside 'medical and scientific purposes' and without a prescription regime. In considering potential contents of an alcohol convention, it is noted that both the FCTC and the drug conventions are concerned at least as much with domestic markets as with international trade. PMID- 17132576 TI - Reducing the burden of smoking world-wide: effectiveness of interventions and their coverage. AB - Cigarette smoking and other tobacco use imposes a huge and growing public health burden globally. Currently, approximately 5 million people are killed annually by tobacco use; by 2030, estimates based on current trends indicate that this number will increase to 10 million, with 70% of deaths occurring in low- and middle income countries. Numerous studies from high-income countries, and a growing number from low- and middle-income countries, provide strong evidence that tobacco tax increases, dissemination of information about health risks from smoking, restrictions on smoking in public places and in work-places, comprehensive bans on advertising and promotion and increased access to cessation therapies are all effective in reducing tobacco use and its consequences. Despite this evidence, tobacco control policies have been unevenly applied--due partly to political constraints. This paper provides a summary of these issues, beginning with an overview of trends in global tobacco use and its consequences and followed by a review of the evidence on the effectiveness of tobacco control policies in reducing tobacco use. A description of the types and comprehensiveness of policies currently in place and a discussion of some of the factors correlated with the strength and comprehensive of these policies follows. PMID- 17132577 TI - A review of the efficacy and effectiveness of harm reduction strategies for alcohol, tobacco and illicit drugs. AB - Harm reduction is both a policy approach and used to describe a specific set of interventions. These interventions aim to reduce the harms associated with drug use. Employing a strict definition of harm reduction, evidence for the efficacy and effectiveness of alcohol, tobacco and illicit drug harm reduction interventions were reviewed. Systematic searches of the published literature were undertaken. Studies were included if they provided evaluation data (pre-post, or control group comparisons). More than 650 articles were included in the review. The majority of the literature concerned illicit drugs. For alcohol, harm reduction interventions to reduce road trauma are well-founded in evidence. Otherwise, there is limited research to support the efficacy and effectiveness of other alcohol harm reduction interventions. For tobacco, the area is controversial but promising new products that reduce the harms associated with smoking are being developed. In the area of illicit drugs there is solid efficacy, effectiveness and economic data to support needle syringe programmes and outreach programmes. There is limited published evidence to date for other harm reduction interventions such as non-injecting routes of administration, brief interventions and emerging positive evidence for supervised injecting facilities. There is sufficient evidence to support the wide-spread adoption of harm reduction interventions and to use harm reduction as an overarching policy approach in relation to illicit drugs. The same cannot be concluded for alcohol or tobacco. Research at a broad policy level is required, especially in light of the failure by many policy makers to adopt cost-effective harm reduction interventions. PMID- 17132578 TI - Civil society-a leader in HIV prevention and tobacco control. AB - Many civil society organisations (CSOs) have been at the forefront of identifying new ideas and implementing innovative models regarding health and health systems around the world. Their activities become highly charged, however, when they engage in advocacy efforts designed to influence change in policies and systems linked with more controversial or complicated public health issues. Policies, laws and regulations regarding illicit drugs and tobacco fall directly into that category. There is no doubt that the use of both kinds of substances can have the same health consequences-including ill health and death-yet they are approached in widely different ways. Smoking is legal to some extent in every country in the world, and is generally considered a matter of personal choice. Many people believe that efforts to limit tobacco use are coercive and impede on individual rights. Those who use illicit drugs such as heroin, meanwhile, are with few exceptions considered social deviants, misfits and lawbreakers. Many CSOs support comprehensive, government-funded prevention strategies coupled with non-punitive, non-judgmental programmes designed to help users change behaviour. Such strategies are designed to reflect and respond to the medically addictive nature of both tobacco and many illegal drugs. Proponents argue that not only are the public health benefits of expansive, well-conceived interventions potentially vast, but so too are the social and economic benefits accruing from lower rates of debilitating disease and premature death. To that end, many international, national and local CSOs are identifying the direct and indirect health consequences of tobacco and illegal drug use; proposing and advocating for strategies to limit their impact; and sharing information and resources with like minded organisations elsewhere. This leadership role has helped influence and shape policy, especially in recent years. This paper examines civil society's involvement in efforts to change drug and tobacco policy in selected countries in Central and Eastern Europe and the former Soviet Union (CEE/FSU). It concludes that in Poland and Kazakhstan, in terms of tobacco control, and increasingly in Ukraine and parts of Central Asia in terms of harm reduction, multi-sectoral approaches are the most effective way to engage citizens and to implement comprehensive strategies to change behaviour by supportive measures, not punitive ones. PMID- 17132579 TI - Community-based interventions and alcohol, tobacco and other drugs: foci, outcomes and implications. AB - The social, health and economic burdens from alcohol, tobacco and other drugs have impacts globally, national and locally. Effective interventions are needed at each level in order to reduce the extensive harm and attendant costs. This paper examines four topics: options available to the local community, evidence of effectiveness, links between local experiences and national and regional initiatives and implications for future research and intervention. It appears that there are a substantial number of options available at the local level. However, evaluation of them is not standard practice, and the results of the higher quality evaluations indicate that many, but not all, interventions have modest or equivocal impact. There is also not a consistent relationship between local and national interventions, although some themes are apparent: in tobacco control there may be good synergy across jurisdictional levels, for alcohol there is evidence that as national control measures are eroded local communities are encouraged or required to take up these agendas, and with regard to illicit drugs there may be tension between law enforcement priorities at the national level and harm reduction orientations locally. Future initiatives need to have appropriate evaluations as a standardised part of prevention initiatives, and include the development of national databases of what is going on locally. These initiatives should promote national policies that include setting parameters and guidelines, but nevertheless do not dictate specific steps and strategies how to achieve local goals in reducing risk and harm. PMID- 17132580 TI - Drug production, trafficking and trade in Asia and Pacific Island countries. AB - We report here on illicit drug production, trafficking and transit routes found in the Asia Pacific region. The report is based on the 'Situational analysis of illicit drug issues and responses in Asia and the Pacific', commissioned by the Australian National Council on Drugs Asia Pacific Drug Issues Committee. The situational analysis was a comprehensive desk based review; data sources included published and unpublished literature and key informant reports. It was found that Myanmar was the main producer of opium, heroin and amphetamine-type stimulants (ATS) in the Asia-Pacific region. China is now considered a major producer of methamphetamines, but other Asia-Pacific nations are also involved in production. Cannabis production was found throughout most of the Asia-Pacific region, in particular Cambodia and the Philippines. Drug trafficking and transit routes of Asia and the Pacific were proliferating and dynamic. The Pacific is mainly known as a trans-shipment point for drugs entering other countries in the region. Drug cultivation and production in Asia is substantial. The expansion of ATS production in the Asia Pacific region is causing much concern. Most drug traffickers change routes and tactics to exploit available vulnerable points along international borders. Responding effectively to the complexity and scale of drug production and trafficking in the Asia-Pacific region will remain a major challenge. PMID- 17132582 TI - The need for change in UK mental health services: South Asian service users' views. AB - OBJECTIVES: UK literature on mental health services for ethnic minority service users relies heavily on perceptions of professionals, carers and community representatives. This research investigates the views of South Asian service users themselves about experiences of mental health services and how they might be improved. DESIGN: Thematic analysis of material from focus groups and individual interviews with Asian mental health service users within one local area. RESULTS: South Asian service users clearly identify the impact of socio economic exclusion upon their mental health. Cultural and institutional exclusion compound this, leading to continuing insensitivity towards their particular needs within hospital and community-based services. Asian service users feel unsafe to share their particular concerns within many service settings. They see advocacy that recognises their experience of exclusion as a significant resource for mental health improvement. They want sounder financing of culturally appropriate services for recovery; further development of the cultural competence of staff within mainstream services; and educational programmes about mental health directed at minority communities. CONCLUSION: UK mental health services remain unresponsive to the consistently expressed views of South Asian service users. A major cultural change is required if the UK Government initiative Delivering Race Equality is to impact successfully at the local level. It will have a greater chance of success if the rhetoric of user involvement is matched by systematic consultation with South Asian service users. PMID- 17132583 TI - Acculturation and body image perception among Latino youth. AB - OBJECTIVES: Examine the prevalence and predictors of body image dissatisfaction and disordered eating among Latino youth. DESIGN: One hundred and sixty-seven Latino youth were surveyed as part of a tailored nutrition communication intervention. The youth's mean age was 12.81 years (SD = 2.74) and 54% were female. Mean self-rated health was 2.59 (SD = 1.02; range: 1 = good to 5 = poor), despite 70% reporting a desire to be thinner. Using age and gender-specific growth charts, 16% of the youth were classified as at risk for overweight and 34% were classified as overweight. RESULTS: Among adolescents, girls (p < or = 0.001), youth who were classified as at risk for or being overweight (p < or = 0.001), and who more strongly recognized and agreed with socially sanctioned standards of appearance as represented in the media (Standardized beta St. beta = 1.86, p < or = 0.001) were more dissatisfied with their body image (R(2) = 0.57). Among children, being at risk for or overweight (p < or = 0.001), reporting a stronger affiliation with the Mexican culture (St. beta = 0.84, p < or = 0.01) and stronger expectations that a healthy diet was associated with improved appearance (St. beta = 0.63, p < or = 0.05) predicted greater body image dissatisfaction (R(2) = 0.55). CONCLUSION: Interventions that address sociocultural attitudes toward appearance may be effective at reducing both the prevalence of body image dissatisfaction and disordered eating. PMID- 17132584 TI - An empirical evaluation of social support and psychological well-being in older Chinese and Korean immigrants. AB - OBJECTIVES: To examine, among older Chinese and Korean immigrants: (1) the concept and measurement of perceived social support; (2) levels of social support and psychological well-being by living arrangement; and (3) whether social support is positively related to psychological well-being. DESIGN: A convenience sample of 200 self-identified Chinese and Koreans, aged 65 years and older, were interviewed. Psychometric analysis was conducted to examine the reliability and validity of a new social support measure. Bivariate and multivariate analyses were used to examine the relationship between social support and psychological well-being. RESULTS: Factor analysis revealed a four-factor solution of social support and adequate psychometrics of all social support scales was found. Multivariate results suggest that Koreans have more depression than Chinese (p < 0.01). Those who lived with their spouse and adult children had lower overall psychological well-being (p < 0.05) and lower positive affect (p < 0.05) compared to those who lived alone. Having more emotional/companionship support significantly (p < 0.001) contributed to better overall psychological well-being, having less depression and higher positive affect. CONCLUSIONS: A new multidimensional social support measure for use with older Chinese and Koreans could be useful upon further validation. These results suggest that older Chinese and Koreans' psychological well-being may be negatively affected when they live with their adult children. More depression in Koreans may be related to difficulties in expanding their social network beyond Korean-speaking people. PMID- 17132585 TI - A cross-national comparison of youth risk behaviors in Latino secondary school students living in El Salvador and the USA. AB - OBJECTIVES: As Latin Americans' exposure to the USA increases through migration patterns and US political and economic ties to their countries of origin, they become susceptible to adopting not only the cultural expressions of the USA such as fashion, but also the health-related behaviors of the US population. In assessing potential health risks for Salvadoran youth that may result from the connection between Latin Americans and the USA, this study compared the prevalence of health risk behaviors from four behavior domains (aggression and victimization, depression and suicidal ideation, substance use, and sexual behavior) between Salvadoran and US Latino secondary school students aged 14-17 years. DESIGN: A secondary analysis was performed on two 1999 cross-sectional survey data. In the USA, results were based on 1,063 Latino high school students who answered the nationally representative Youth Risk Behavior Survey (YRBS) conducted by the Centers for Disease Control and Prevention. In El Salvador, results were based on 793 public secondary school students who answered a local YRBS survey conducted in coordination with the Ministry of Education of El Salvador. RESULTS: The prevalence rates for aggression/victimization and for depression and suicidal ideation behaviors were similar between Salvadoran and US Latino adolescents. Substance use prevalence, however, was 10-40% higher for US Latino adolescents. While the prevalence of sexual intercourse was higher among US Latino youth (between 13 and 27% higher, depending on age), the prevalence of condom use was lower among sexually active Salvadoran youth (between 11 and 42% lower, depending on age). CONCLUSIONS: In the context of the transnationalization of the Salvadoran population, with potential for increased influence of the USA in Salvadoran culture, these differences in risk behavior are important for targeting effective interventions for Latino adolescents in El Salvador and in the USA. PMID- 17132587 TI - The need for a European strategy on global health. PMID- 17132586 TI - Indigenous and non-indigenous child oral health in three Australian states and territories. AB - OBJECTIVES: To explore the prevalence and severity of Indigenous and non Indigenous child dental disease in relation to age, sex, residential location and socio-economic status in three Australian states and territories. DESIGN: Children aged 4-14 years who were enrolled in a school dental or screening service in New South Wales, South Australia and the Northern Territory, Australia, were randomly selected to take part in this cross-sectional study. Bivariate and multivariate analyses were used to assess outcomes. RESULTS: A total of 328,042 children were included, of which 10,517 (3.2%) were Indigenous. Some 67.1% of Indigenous children lived in rural areas and 47.3% lived in areas of high disadvantage. About 37.5% of 4- to 10-year-old Indigenous children had no experience of dental disease in the primary dentition while 70.7% of 6- to 14 year-old Indigenous children had caries-free permanent dentitions. The mean number of decayed, missing and filled primary teeth (dmft) of Indigenous 4- to 10 year-old children was 2.9 (SD; 3.4) while the mean DMFT of Indigenous 6- to 14 year-old children was 0.8 (SD; 1.6). Across all age-groups, Indigenous children living in the most deprived areas had higher dmft and DMFT levels than their more socially advantaged counterparts, while rural-dwelling Indigenous children had higher levels of dental disease experience than metropolitan-dwelling Indigenous children. After adjusting for potential confounding, Indigenous children aged 4 10 years were over twice as likely to have caries in the deciduous dentition than similarly aged non-Indigenous children (OR: 2.25, CI: 2.14-2.36), and 6- to 14 year-old Indigenous children were over one and a half times more likely to have decay in the permanent dentition (OR: 1.68, CI: 1.60-1.77) than their non Indigenous counterparts. CONCLUSION: Indigenous children experienced higher caries prevalence and severity than non-Indigenous children, irrespective of other socio-demographic factors. Factors concerning Indigenous social capital may have influenced our findings. PMID- 17132588 TI - Assessing the quality of Swedish health economics research. PMID- 17132589 TI - Effects of body fatness and physical activity on cardiovascular risk: risk prediction using the bioelectrical impedance method. AB - AIMS: To explore the effects of body fat percentage (BF%) on incidence of and mortality from cardiovascular disease (CVD) and to study the cardio-protective effect of physical activity in relation to BF%. METHODS: A total of 26,942 men and women, aged 45-73 years, without history of CVD were followed up for incidence of coronary events (CE), ischaemic stroke, and CVD mortality over seven years in relation to sex-specific quartiles (Q1-Q4) of BF%. The cardio-protective effect of leisure-time physical activity was studied in relation to BF%. RESULTS: In men, the relative risk (RR) for CE and CVD mortality increased progressively with BF%. RR for CE in Q4 was 1.37 (95% confidence interval: 1.07-1.74), adjusted for age, height, smoking, high alcohol intake, and physical activity, compared with Q1. In women, BF% was significantly associated with incidence of CE and stroke. BF% was more strongly correlated to body mass index (BMI) (r = 0.83) and waist circumference (r = 0.76) in women than in men (r = 0.59 and r = 0.66, respectively). BF% was a stronger risk factor than BMI in women, and equally strong as waist circumference. A significant interaction (p = 0.013 for incidence of CE, p = 0.026 for ischaemic stroke) was found between BF% and sex. The raised cardiovascular risk was reduced by physical activity in subjects with high BF%. CONCLUSIONS: BF% is a risk factor for CE, ischaemic stroke, and CVD mortality. An interaction between BF% and sex suggests that BF% is a stronger CVD risk factor in women. The raised cardiovascular risk associated with high BF% is reduced by physical activity. PMID- 17132590 TI - Contribution of Swedish moist snuff to the metabolic syndrome: a wolf in sheep's clothing? AB - AIM: Combined effects of genetic and environmental factors underlie the clustering of cardiovascular risk factors in the metabolic syndrome (MetSy). The aim was to investigate associations between several lifestyle factors and MetSy, with a focus on the possible role of smokeless tobacco in the form of Swedish moist snuff (snus). METHODS: A population-based longitudinal cohort study within the Vasterbotten Intervention Programme in Northern Sweden. All inhabitants at the ages of 30, 40, 50, and 60 are invited to participate in a health survey that includes a questionnaire on psychosocial conditions and lifestyle and measurement of biological variables. Individuals examined in 1990-94 (n = 24,230) and who also returned for follow-up after 10 years were included (total of 16,492 individuals: 46.6% men and 53.4% women). Regression analyses were performed. MetSy was the outcome and analyses were adjusted for age, sex, alcohol abuse, and family history of CVD and diabetes. RESULTS: Ten-year development of MetSy was associated with high-dose consumption of snus at baseline (OR 1.6 [95% CI 1.26 2.15]), low education (2.2 [1.92-2.63]), physical inactivity (1.5 [1.22-1.73]) and former smoking (1.2 [1.06-1.38]). Snus was associated with separate components of MetSy, including triglycerides (1.6, 1.30-1.95), obesity (1.7 [1.36 2.18]) but not hypertension, dysglycemia and low HDL cholesterol. CONCLUSIONS: MetSy is independently associated with high consumption of snus, even when controlling for smoking status. The finding is of public health interest in societies with widespread use of snus. More research is needed to better understand the mechanisms underlying this effect. PMID- 17132591 TI - More illness and less disease? A 20-year perspective on chronic disease and medication. AB - BACKGROUND: The implication of medication is that a drug is given against an illness. Over the last few decades an expanding number of drugs have appeared that focus on reducing risk factors and lifestyle conditions. AIMS: To investigate the apprehension in respect of chronic disease among the working population in a Swedish community in 2000 compared with 1980. METHODS: In 1980 and 2000 an analogous questionnaire assessing chronic diseases and medication was mailed to 250 randomly selected persons from a local population between 25 and 70 years of age. RESULTS: Some 80% of the persons (n = 201) replied in 1980 and 78% (n = 195) in 2000. Individuals declaring a chronic disease increased from 23% (46/201) in 1980 to 39.5% in 2000 (p = 0.0005). Corresponding figures for men were 20.6% (20/97) in 1980 and 40.8% (40/98) in 2000 (p = 0,004) and for women 25.0% (26/104) in 1980 and 38.1% (37/97) in 2000 (p = 0.064). Persons who regularly see a doctor increased from 13% to 26% (p = 0.002) and the use of drugs for chronic diseases increased from 19% to 33% (p = 0.002). In 2000 an average of 2.3 drugs per person were used among those with a chronic disease, an increase of 53% since 1980. CONCLUSION: Stated chronic diseases and use of drugs for such diseases increased greatly between1980 and 2000. Prescribing drugs for a "risk" with no apparent illness may be confused with the remedy for an illness. PMID- 17132592 TI - Risk-factor profile for the incidence of subarachnoid and intracerebral haemorrhage, cerebral infarction, and unspecified stroke during 21 years' follow up in men. AB - AIMS: To study the risk-factor profile for the incidence of non-fatal and fatal stroke among middle-aged men according to the stroke subtypes subarachnoid or intracerebral haemorrhage, cerebral infarction, and unspecified stroke. METHODS: The study design is a prospective cohort study. A total of 16,209 men aged 40-49 years resident in Oslo were screened for cardiovascular disease risk factors in 1972-73. Of these, 14,403 men had no cardiovascular symptoms or diseases or diabetes. The incidence of stroke after 21 years of follow-up of all men was extracted from hospital records and linkage to Statistics Norway. RESULTS: A total of 429 non-fatal and 107 fatal stroke events were registered. Case fatality within 28 days (number and percentage of cases) was 51% (41, 7.7%) for subarachnoid haemorrhage, 39% (67, 12.6%) for cerebral haemorrhage, 10% (246, 46.3%) for cerebral infarct, and 19% (177, 33.4%) for unspecified stroke. Risk of stroke (not subarachnoid haemorrhage) increased with the presence of symptoms or a history of cardiovascular disease or diabetes. In multivariate analysis of men without CVD or diabetes, high blood pressure was a risk factor for all subtypes of stroke; furthermore, daily smoking was a risk factor for all subtypes except subarachnoid haemorrhage. Serum cholesterol and glucose concentrations and height (inverse association) were independently associated with cerebral infarction. Smoking was a significantly stronger predictor of fatal than non-fatal events. CONCLUSIONS: The risk-factor profile differed according to the underlying subtype of stroke. Cerebral infarction clearly shared with myocardial infarction the classical risk factors, including non-fasting glucose concentration. PMID- 17132593 TI - Client perceptions of the performance of public and independent not-for-profit primary healthcare. AB - AIMS: To compare primary healthcare (PHC) provided by an independent not-for profit organization (INPO) with that provided by two public municipal organizations (MO1 and MO2), in terms of clients' perceptions of performance, acceptance, and trust. METHODS: A survey using a pre-tested questionnaire to all clients visiting a health centre (HC) doctor or nurse during one week in 2000 (n = 511, 51% response rate) and 2002 (n = 275, 47%). The data were analysed by descriptive statistics and cumulative logistic regression analysis. RESULTS: The INPO differed from both publicly provided services in accessibility, consistency of service, and outcomes. Clients reported lower trust in HC provided by public organizations compared with the INPO. Trust was higher if clients also reported experiencing "very good" or "moderate" organizational access--or if general satisfaction was "very high" or "moderate" or if they experienced outcomes as "very good" or "moderate" compared with the "very poor or low" situation. Women reported lower trust in HC than men. When the family doctor was included in the same logistic regression model with the service provider, only the family doctor was a significant explanatory variable. Reported acceptance of private alternative service providers among clients was similar between the study organizations. CONCLUSIONS: Clients of the INPO generally rated the service more positively than clients of publicly provided services. The results indicate that trust in HC depends more on a family doctor system than a service provider. PMID- 17132594 TI - COHb% as a marker of cardiovascular risk in never smokers: results from a population-based cohort study. AB - AIM: Carbon monoxide (CO) in blood as assessed by the COHb% is a marker of the cardiovascular (CV) risk in smokers. Non-smokers exposed to tobacco smoke similarly inhale and absorb CO. The objective in this population-based cohort study has been to describe inter-individual differences in COHb% in never smokers and to estimate the associated cardiovascular risk. METHODS: Of the 8,333 men, aged 34-49 years, from the city of Malmo, Sweden, 4,111 were smokers, 1,229 ex smokers, and 2,893 were never smokers. Incidence of CV disease was monitored over 19 years of follow up. RESULTS: COHb% in never smokers ranged from 0.13% to 5.47%. Never smokers with COHb% in the top quartile (above 0.67%) had a significantly higher incidence of cardiac events and deaths; relative risk 3.7 (95% CI 2.0-7.0) and 2.2 (1.4-3.5), respectively, compared with those with COHb% in the lowest quartile (below 0.50%). This risk remained after adjustment for confounding factors. CONCLUSION: COHb% varied widely between never-smoking men in this urban population. Incidence of CV disease and death in non-smokers was related to COHb%. It is suggested that measurement of COHb% could be part of the risk assessment in non-smoking patients considered at risk of cardiac disease. In random samples from the general population COHb% could be used to assess the size of the population exposed to second-hand smoke. PMID- 17132595 TI - Association between fruit and vegetable consumption and birth weight: a prospective study among 43,585 Danish women. AB - OBJECTIVE: To examine whether fruit and vegetable consumption in pregnancy is associated with birth weight in a Western population. DESIGN: Prospective cohort study based on telephone interviews, a food frequency questionnaire (FFQ), and extractions of birth characteristics from national health registries. SUBJECTS AND SETTING: The 43,585 Danish women from the Danish National Birth Cohort who had completed the FFQ in mid-pregnancy and on whom information about birth outcome was available. The exposures were frequency of green leafy vegetable (GLV) intake and quantified intake of fruit, fruit and vegetables, and fruit and vegetables and juice. The outcomes were birth weight and z-score for expected birth weight adjusted for sex and gestation week. Information on maternal height, weight, smoking, and other potential confounders was obtained through telephone interviews. RESULTS: Significant associations were found for all exposures to fruit and vegetable intake with birth weight and most with z-score. The strongest association was found for fruit intake in which case birth weight increased by 10.7 g (95% CI 7.3-14.2) per quintile. All associations were stronger among lean women (BMI<20, n = 7,169), whose children's birth weight increased by 14.6 g (95% CI 6.4-22.9) per quintile increase in fruit intake. For GLV the results were more inconclusive. When adjusted for confounders, but not for energy, the association between GLV and birth weight was significant, but the same was not the case for z score. CONCLUSION: Fruit and vegetable consumption in pregnancy is positively associated with birth weight in well-nourished Danish women, especially among lean women. PMID- 17132596 TI - Representativeness in population-based studies: a detailed description of non response in a Danish cohort study. AB - BACKGROUND: Decreasing rates of participation in population-based studies increasingly challenge the interpretation of study results, in both analytic and descriptive epidemiology. Consequently, estimates of possible differences between participants and non-participants are increasingly important for the interpretation of study results and generalization to the background population. METHODS: An age-specific, population-based cohort of 1,198 individuals was examined at age 40, 45, 51, and 60. Participants were compared with non participants and when possible also with the background population using a wide range of detailed information on somatic and mental health collected at each examination, including data from a clinical examination, biochemical measurements, questionnaires, interviews, and public registers. RESULTS: Participation rates were higher than 80% at examinations at age 40, 45, and 51, but decreased to 65% at age 60. At the baseline investigation at age 40, analyses indicated that participants were representative of the cohort as well as the background population. However, the mortality rate was higher among non participants in the succeeding 20 years. Among living cohort members at the 60 year examination, non-participants had lower socioeconomic status, higher hospitalization rate, and a worse overall health profile than participants. CONCLUSIONS: The detailed data presented reinforce the contention that the health profile of non-participants is typically worse than that of participants. The results also indicate that while data from public registers give easily accessible information about non-participants, these crude proxy measures of health may not be enough to document representativeness. PMID- 17132597 TI - Rural-urban differences in health and health behaviour: a baseline description of a community health-promotion programme for the elderly. AB - STUDY OBJECTIVE: To (1) describe the setting and design of the Good Ageing in Lahti Region (GOAL) programme; (2) by using the baseline results of the GOAL cohort study, to examine whether living in urban, semi-urban, or rural communities is related to risk factors for chronic diseases and functional disability in ageing individuals. DESIGN: The baseline data of a cohort study of ageing individuals living in three community types (urban, semi-urban, rural). Data were collected by two questionnaires and laboratory assessments. SETTING: Fourteen municipalities in the Lahti region (Paijat-Hame County) in Finland. PARTICIPANTS: A regionally and locally stratified random sample of men and women born in 1946-50, 1936-40, and 1926-30. A total of 4,272 were invited and 2,815 (66%) participated. MAIN RESULTS: Elevated serum cholesterol, obesity, disability, sedentary lifestyle (<2 times/week walking), and high fat intake were more prevalent in rural vs. urban and semi-urban communities. After adjustment for sex, age, education, obesity, diet, physical activity, smoking, and alcohol use, rural communities remained the only community type with increased (p<0.05) probability for high BMI (OR 1.33) and high waist circumference (OR 1.43). CONCLUSIONS: The unfavourable health and lifestyle profile, together with an old population, makes health promotion for elderly citizens a special challenge for rural communities such as those in Paijat-Hame County, Finland. Most, if not all, of the differences in health between the three community types were explained by educational background, physical activity, and smoking. PMID- 17132598 TI - Cost-effectiveness of healthcare-based interventions aimed at improving physical activity. AB - AIM: This article aims to review current knowledge concerning the cost effectiveness of healthcare-based interventions aimed at improving physical activity. METHOD: A search was performed for economic evaluations containing the terms "physical activity", "exercise", or "fitness". Cost-effectiveness for the articles found was described based on a model for evaluating interventions intended to promote physical activity. RESULTS: A total of 26 articles were found in the search. Nine of them concern a general population, 7 evaluated older people, and 10 studied disease-specific populations. A preventive perspective is most common, but some have a treatment perspective. Around 20 of the interventions studied were cost-effective according to their authors, but all analyses had some shortcomings in their evaluation methods. CONCLUSION: This review found many examples of cost-effective interventions. There is a lack of evidence for the cost-effectiveness of interventions aimed at those whose only risk factor for illness is a sedentary lifestyle. There is more evidence, although it is limited, for the cost-effectiveness of interventions aimed at high risk groups or those who manifest poor health related to physical inactivity. Most of the evidence for cost-effectiveness is for older people and those with heart failure. Promotion of physical activity can be cost-effective with different methods and in different settings, but there remains a lack of evidence for specific methods in specific populations. PMID- 17132599 TI - Physicians' opinions and experiences of the Pharmaceutical Benefits Reform. AB - AIM: To investigate opinions on and experiences of the new Pharmaceutical Benefits Reform (PBR) among physicians working in the Swedish county council of Region Vastra Gotaland. METHODS: Questionnaires were sent to all private practitioners who had contracts with the county council of Region Vastra Gotaland (n = 320) and a random sample of 25% of the doctors employed by the same county council (n = 1,068). The questionnaire comprised questions on how the physicians had received information on the PBR and sought opinions on the introduction of generic substitution in pharmacies, workplace codes, individual prescriber codes, and the Pharmaceutical Benefits Board. Analyses were performed with logistic regression. RESULTS: The response rate was 65%. The majority of doctors had received sufficient information about the reform, most often from their employer, followed by the Drug and Therapeutics Committees. More than half of the respondents were positive about the introduction of generic substitution, which was associated with the respondent's age, experiences, and employer. Most of the doctors thought that generic substitution could reduce the health system's pharmaceutical expenditure, which was associated with the employer. One-third reported that generic substitution had caused problems for their patients in the past month. The probability of being mainly positive towards the newly created Pharmaceutical Benefits Board, which decides on reimbursement, increased with increasing age. CONCLUSIONS: Most participating doctors had received sufficient information about the reform and thought that generic substitution could save money for society. Age, employer, and experiences appeared to influence opinions on several issues. PMID- 17132600 TI - Smoking in pregnancy in relation to gender and adult mortality risk in offspring: the Helsingborg Birth Cohort Study. AB - BACKGROUND: Smoking in pregnancy is a well-documented risk factor for fetal growth impairment and poor perinatal outcomes. Less is known about the long-term effects of maternal smoking on offspring mortality. METHODS: A follow-up study in national registers on total mortality and cancer based on a birth cohort from Helsingborg, Sweden, including data on 2,010 sons and 1,982 daughters born to mothers for whom the smoking habits during pregnancy (50% smokers) have been recorded. RESULTS: A total of 92 offspring deaths were recorded (54 men, 38 women) during follow-up. Of these deaths, 43 deaths were related to trauma, 6 to circulatory disease, and 2 to endocrine disorders. In men, an elevated mortality risk was associated with increasing maternal smoking habits (p for trend 0.011), but in women with low birth weight (p for trend 0.006). A total of 47 incident offspring cancers were registered (18 in men and 29 in women). No significant relation was noted for maternal smoking habits and cancer in the offspring. CONCLUSIONS: Maternal smoking during pregnancy is associated with an increased mortality risk in early adult life for male offspring but not for female offspring. This could represent the possible consequence of an increased susceptibility in male fetuses. PMID- 17132601 TI - The magnitude of bias in a cross-sectional study on lifestyle factors in relation to Type 2 diabetes. AB - AIM: In cross-sectional studies it may be difficult to ascertain the temporal order of exposure and disease, which may have consequences for causal inference. The authors aimed to illustrate the possible magnitude of this potential bias using data from a cross-sectional study on coffee consumption and work stress in relation to type 2 diabetes. METHODS: By a series of computer simulations the authors examined to what extent the observed negative association between type 2 diabetes and high coffee consumption and positive association between type 2 diabetes and high work stress could be due to reverse causality, by assuming that cases changed their exposures in response to development of the disease. RESULTS: If the negative association between coffee and type 2 diabetes was a consequence of reversed causality, 30-40% of the cases would have to decrease their coffee consumption from 5 cups of coffee per day to 3-4 cups per day and from 3-4 cups per day to 2 cups of coffee per day. Moreover, approximately 60% of the cases would have to increase their work stress from low to medium work stress and from medium to high work stress, in order to produce the positive association with diabetes that was observed. CONCLUSION: Even if the type 2 diabetic patients to some extent may have changed their exposure in response to disease development, it seems unlikely that the associations observed between type 2 diabetes, coffee consumption, and work stress are due to this bias. PMID- 17132602 TI - beta2 adrenoceptor (ADRB2) pharmacogenetics and cardiovascular phenotypes during laryngoscopy and tracheal intubation. AB - Pharmacogenetics is the study of genetic variations that cause a variable drug response characterized by alteration in drug metabolism or in pharmacodynamics. The polymorphisms in genes encoding receptors relevant to treatment cause variation in sensitivity to many drugs. beta2 Adrenoceptor genetic variation contributes to regulation of blood pressure and hemodynamic changes by mediating peripheral vasodilatation. Laryngoscopy and tracheal intubation associated with hemodynamic changes. Although there are four nonsynomic single-nucleotide polymorphisms (SNPs) of beta2 adrenoceptor gene, codon 16 (Arg16Gly) and codon 27 (Gln27Glu) SNPs are both common and functionally important. In this paper, the authors investigated the beta2 adrenoceptor Gly16 and Glu27 SNPs in response to drugs relevant to anesthesia and how these SNPs impacted upon the cardiovascular phenotypes. The authors measured arterial systolic and diastolic blood pressure, heart rate, and rate-pressure product before induction of anesthesia and 1 min following laryngoscopy and tracheal intubation. Genomic DNA was amplified and genotyped using allele-specific polymerase chain reaction (ASPCR) and restriction fragment length polymorphism (RFLP) assays, respectively. When the authors compared hemodynamic results according to genotypes, the patients with Gln homozygote allele at codon 27 exhibited significant increase of heart rate than patients with Glu allele after laryngoscope and tracheal intubation. PMID- 17132603 TI - Toxicity of cerium oxide nanoparticles in human lung cancer cells. AB - With the fast development of nanotechnology, the nanomaterials start to cause people's attention for potential toxic effect. In this paper, the cytotoxicity and oxidative stress caused by 20-nm cerium oxide (CeO2) nanoparticles in cultured human lung cancer cells was investigated. The sulforhodamine B method was employed to assess cell viability after exposure to 3.5, 10.5, and 23.3 microg/ml of CeO2 nanoparticles for 24, 48, and 72 h. Cell viability decreased significantly as a function of nanoparticle dose and exposure time. Indicators of oxidative stress and cytotoxicity, including total reactive oxygen species, glutathione, malondialdehyde, alpha-tocopherol, and lactate dehydrogenase, were quantitatively assessed. It is concluded from the results that free radicals generated by exposure to 3.5 to 23.3 microg/ml CeO2 nanoparticles produce significant oxidative stress in the cells, as reflected by reduced glutathione and alpha-tocopherol levels; the toxic effects of CeO2 nanoparticles are dose dependent and time dependent; elevated oxidative stress increases the production of malondialdehyde and lactate dehydrogenase, which are indicators of lipid peroxidation and cell membrane damage, respectively. PMID- 17132604 TI - Melatonin protection against lead-induced changes in human neuroblastoma cell cultures. AB - The nervous system is the primary target for low-levels of lead (Pb) exposure and the developing brain appears to be especially vulnerable to Pb neurotoxicity. Chronic low-level Pb exposure causes growth retardation and intellectual impairment. In the present study the protective effect of melatonin during exposure to low-levels of Pb in human SH-SY5Y neuroblastoma cell cultures was assessed. The cells were exposed to Pb (0.01 to 10 microM) for 48 h. Pb inhibited the proliferation of neuroblastoma cells significantly in a concentration dependent manner. A 50% inhibition (IC50) of cell proliferation was observed at about 5 microM Pb. Pb decreased (16% to 62%) the levels of total cellular glutathione (GSH) in a concentration (0.1 to 10 microM)-dependent manner. Exposure of cells to Pb (5 microM) for 48 h resulted in an eightfold increase in caspase-3 activity and prostaglandin E2 (PGE2) level. Pretreatment with melatonin (10 microM) blocked the effects of Pb on GSH content and caspase-3 activity, and showed significant improvement in reducing the level of PGE2. The results suggest that some of the neurotoxic effects of Pb may be partly mediated by apoptosis and pretreatment with melatonin can prevent these effects. The present study asserts the neuroprotective effect of melatonin in conditions of Pb-induced toxicity in neuroblastoma cell cultures. PMID- 17132605 TI - Acute and subchronic toxicity studies on Sel-Plex, a standardized, registered high-selenium yeast. AB - Selenium has been recognized as an essential nutrient for human health; however, its bioavailability is primarily dependent upon the type of selenium, elemental versus organic. In geographic areas low in selenium, there is the potential for animals (including humans) to become selenium deficient and this potential deficiency can be remedied by consumption of exogenous selenium, including selenium-enriched yeast (Saccharomyces cerevisiae) that contains high levels of organic selenium (e.g., selenized yeast). The present studies were conducted to investigate potential oral toxicity of a unique selenized yeast preparation (Sel Plex) when administered to (1) adult female CHS Swiss mice ICo:OFI (IOPS Caw); (2) adult female CHS Sprague-Dawley rats; and (3) adult male and female Sprague Dawley CD rats. For the 28- and 90-day toxicity studies, (1) adult male and female Sprague-Dawley CRL:CD(R)(SD) IGS BR strain rats and (2) adult male and female 6- to 7-month-old Beagle dogs were used. The LD50 for mice was >or=2000 mg Sel-Plex/kg (>or=4.06 mg Se/kg) and for rats, was greater than >or=2000 mg Sel Plex/kg (>or=4.06 mg Se/kg). In the two 28-day studies, for rats, the no observed adverse effects level (NOAEL) was 50 mg Sel-Plex/kg/day (0.1 mg Se/kg/day), and for the dogs, the NOAEL was 22.5 mg Sel-Plex/kg/day (0.045 mg Se/kg/day). For the two 90-day studies, for rats the NOAEL for Sel-Plex was 114 mg/kg/day (0.23 mg Se/kg/day), and for dogs, the NOAEL was 30 mg Sel-Plex/kg/day (0.06 mg Se/kg/day): the latter being the NOAEL in the most sensitive species. PMID- 17132606 TI - Genotoxicity studies on Sel-Plex, a standardized, registered high-selenium yeast. AB - Selenium, recognized as an essential nutrient for human health, is a component of proteins and enzymes required for various biological functions and is currently being used as a feed supplement for livestock in geographical areas that are naturally low in selenium. Selenium is structurally similar to sulfur, replacing the sulfur atom in stoichiometric amounts and thus functions through an association with proteins, termed selenoproteins. In geographic areas low in selenium, there is the potential for animals (including humans) to become selenium deficient and this potential deficiency can be remedied by consumption of exogenous selenium, including selenium-enriched yeast (Saccharomyces cerevisiae) that contains high levels of organic selenium (e.g., selenized yeast). A unique, standardized, registered high selenium food-grade baker's yeast (S. cerevisiae; Sel-Plex), was tested in the following battery of Genotoxicity assays; (1) a bacterial reverse mutation test (Ames test); (2) an in vitro mammalian chromosome aberration test; and (3) a mouse micronucleus test. Under the conditions of this assay, Sel-Plex showed no evidence of mutagenic activity in Salmonella typhimurium, in the bacterial reverse mutation test. Sel-Plex did not induce significant chromosomal aberrations in cultured human lymphocytes in the in vitro mammalian chromosome aberration test. Sel-Plex did not statistically increase the frequency or proportion of micronucleated immature erythrocytes in the mouse micronucleus test. Thus, from the studies presented here, the authors conclude that Sel-Plex is nongenotoxic. PMID- 17132607 TI - Comparative immunotoxicity evaluation of amphotericin B and ABELCET, an amphotericin B lipid complex (ABLC). AB - ABELCET, an amphotericin B lipid complex formulation (ABLC) and an aqueous, non lipid-containing formulation with sodium deoxycholate (AmBd), were evaluated for their potential to induce immunotoxicity in B6C3F1 female mice. ABLC was administered intravenously at doses of 1, 3, and 10 mg/kg daily for 28 days, while AmBd at 1 mg/kg was administered by the same route and duration. The effect of ABLC and AmBd on clinical signs, body weight, and spleen weight was determined. Peritoneal macrophage function was measured by phagocytosis of 51Cr labeled chicken red blood cells and generation of hydrogen peroxide during respiratory burst. The ability of natural killer cells to lyse radiolabeled tumor target cells was evaluated in a short-term chromium-release assay. The ability of splenic T and B cells to undergo blastogenesis and of splenic T cells to recognize alloantigens present on foreign cells was assessed in a splenic lymphocyte assay and the ability of mice to generate antibody-forming cells following immunization with sheep red blood cells was measured. Neither ABLC nor AmBd affected the metabolic or functional activity of murine phagocytic cells. These agents also did not cause any biologically significant or dose-related changes in B- or T-cell responses to mitogens, T-cell responses to allogeneic cells in the mixed lymphocyte culture assay, or natural killer cell function. The ability to generate a primary antibody response to a T cell-dependent antigen was also unimpaired. Based on the results of this study, it was concluded that neither ABLC at dose up to 10 mg/kg nor AmBd at dose up to 1.0 mg/kg produce biologically significant immunologic changes in B6C3F1 mice. PMID- 17132608 TI - Chronic contamination with 137cesium in rat: effect on liver cholesterol metabolism. AB - After the Chernobyl nuclear accident, epidemiological studies on human populations living in 137Cs-contaminated areas revealed the increase frequencies of thyroid cancer and evoked the apparition of cardiovascular diseases, hormonal effect, liver alteration, and lipid disorder. Actually, it raises a problem of public safety for the populations living on these territories that are exposed to low levels of 137Cs during a long period through food. Then it is necessary to study potential effect of this chronic contamination. To mimic this situation, the authors investigate the potential biological effects of chronic exposure to 137Cs at a postaccidental dose (150 Bq/rat/day) on hepatic metabolism of cholesterol in rat. Plasma lipid level, gene expression and activity were analyzed. It was observed that in 137Cs-exposed rats, gene expression of low density lipoprotein receptor (LDLr), apolipoprotein B (apoB), and liver X receptor alpha (LXRalpha) are increased (95%, p < .05; 34%, p < .05; 20%, p < 0.05, respectively), whereas transporter adenosine triphosphate-binding cassette transporter G5 (ABCG5) is decreased (42%, p < .05). In addition, cytochrome P450 27A1 (CYP27A1) activity is increased (34%, p < .05) in contaminated rat liver. In conclusion, the results suggest that 137Cs contamination at low-level induces molecular modifications of the liver cholesterol metabolism without leading to a dysregulation of its homeostasis. These results suggest that chronic long term exposure at low-level of 137Cs may evolve to lipid disorder. PMID- 17132609 TI - Nonclinical vehicle use in studies by multiple routes in multiple species. AB - The laboratory toxicologist is frequently faced with the challenge of selecting appropriate vehicles or developing utilitarian formulations for use in in vivo nonclinical safety assessment studies. Although there are many vehicles available that may meet physical and chemical requirements for chemical or pharmaceutical formulation, there are wide differences in species and route of administration specific to tolerances to these vehicles. In current practice, these differences are largely approached on a basis of individual experience as there is only scattered literature on individual vehicles and no comprehensive treatment or information source. This approach leads to excessive animal use and unplanned delays in testing and development. To address this need, a consulting firm and three contract research organizations conducted a rigorous data mining operation of control (vehicle) data from studies dating from 1991 to present. The results identified 65 single component vehicles used in 368 studies across multiple species (dog, primate, rat, mouse, rabbit, guinea pig, minipig, chick embryo, and cat) by multiple routes. Reported here are the results of this effort, including maximum tolerated use levels by species, route, and duration of study, with accompanying dose limiting toxicity. Also included are basic chemical information and a review of available literature on each vehicle, as well as guidance on volume limits and pH by route and some basic guidance on nonclinical formulation development. PMID- 17132610 TI - The protective effect of melatonin against fumonisin-induced renal damage in rats. AB - The present study was designed to investigate the potential protective effect of melatonin against the renal toxicity of fumonisin in female rats. Six groups of animals were used in this study. The first group served as control. The second group was given melatonin only at a dose level of 10 mg/kg. The third group was fed ration contaminated with fumonisin (100 mg/kg diet). The fourth group was fed ration contaminated with fumonisin (200 mg/kg diet). The fifth group was given daily interperitoneal injection (IP) 10 mg/kg melatonin and fed ration contaminated with fumonisin (100 mg/kg diet). The sixth group was given daily interperitoneal injection of 10 mg/kg melatonin and fed ration contaminated with fumonisin (200 mg/kg diet). The rats were treated for 1 month. Histopathological and histochemical changes in the kidney were investigated. In addition, DNA ploidy was measured in the kidney. Fumonisin administration (100 or 200 mg/Kg diet) to unpretreated control rats caused extensive renal damage as evaluated by histopathology, histochemistry, and/or DNA ploidy measurement. No apparent changes following administration of melatonin. Melatonin coadministration to the fumonisin-administered rats reduced kidney damage and the tissues appeared more or less like the normal. The present study indicates that melatonin has a protective effect in fumonisin-induced renal damage. PMID- 17132611 TI - Lack of trigeminal nerve toxicity in rats exposed to trichloroethylene vapor for 13 weeks. AB - Male and female Fischer-344 rats were exposed to 1,1,2-trichloroethylene (TCE) at 250, 800, or 2500 ppm for 6 h/day, 5 days/week, for 13 weeks. Weekly body weights and daily clinical observations were recorded and a functional observational battery (FOB) was performed monthly. Postexposure neurotoxicological evaluations included an electrodiagnostic evaluation of auditory function, the trigeminal nerve, and a comprehensive neuropathological examination. After 8 weeks of exposure, female, but not male, rats exposed to 2500 ppm were slightly more reactive to handling than the controls but not after 13 weeks of exposure. After 13 weeks, female rats exposed to 2500 ppm TCE were slightly more active during the 1-min observation period than the controls. There were no treatment-related differences in grip performance, landing foot splay, or on the trigeminal nerve evoked potential at any dose. At 2500 ppm TCE, mild frequency-specific hearing deficits were observed, including elevated tone-pip auditory brainstem response thresholds. Focal loss of hair cells in the upper basal turn of the cochlea was observed in 2500 ppm-exposed rats. Except for the cochleas of 2500 ppm-exposed rats, no treatment-related lesions were noted during the neuro-histopathologic examination. The no-observable-adverse-effect level for this study was 800 ppm based on ototoxicity at 2500 ppm. PMID- 17132614 TI - Numerical modeling of 1D arterial networks coupled with a lumped parameters description of the heart. AB - The investigations on the pressure wave propagation along the arterial network and its relationships with vascular physiopathologies can be supported nowadays by numerical simulations. One dimensional (1D) mathematical models, based on systems of two partial differential equations for each arterial segment suitably matched at bifurcations, can be simulated with low computational costs and provide useful insights into the role of wave reflections. Some recent works have indeed moved in this direction. The specific contribution of the present paper is to illustrate a 1D numerical model numerically coupled with a model for the heart action. Typically, the action of the heart on the arterial system is modelled as a boundary condition at the entrance of the aorta. However, the left ventricle (LV) and the vascular network are a strongly coupled single mechanical system. This coupling can be relevant in the numerical description of pressure waves propagation, particularly when dealing with pathological situations. In this work, we propose a simple lumped parameter model for the heart and show how it can be coupled numerically with a 1D model for the arteries. Numerical results actually confirm the relevant impact of the heart-arteries coupling in realistic simulations. PMID- 17132615 TI - Simulation of deformable models with the Poisson equation. AB - In this paper, we present a new methodology for the deformation of soft objects by drawing an analogy between the Poisson equation and elastic deformation from the viewpoint of energy propagation. The potential energy stored due to a deformation caused by an external force is calculated and treated as the source injected into the Poisson system, as described by the law of conservation of energy. An improved Poisson model is developed for propagating the energy generated by the external force in a natural manner. An autonomous cellular neural network (CNN) model is established by using the analogy between the Poisson equation and CNN to solve the Poisson model for the real-time requirement of soft object deformation. A method is presented to derive the internal forces from the potential energy distribution. The proposed methodology models non linear materials with the non-linear Poisson equation and thus non-linear CNN, rather than geometric non-linearity. It not only deals with large-range deformations, but also accommodates isotropic, anisotropic and inhomogeneous materials by simply modifying constitutive coefficients. A haptic virtual reality system has been developed for deformation simulation with force feedback. Examples are presented to demonstrate the efficiency of the proposed methodology. PMID- 17132616 TI - Development and validation of a generic 3D model of the distal femur. AB - The development and validation of a virtual generic 3D model of the distal femur using computer graphical methods is presented. The synthesis of the generic model requires the following steps: acquisition of bony 3D morphology using standard computed tomography (CT) imaging; alignment of 3D models reconstructed from CT images with a common coordinate system; computer graphical sectioning of the models; extraction of bone contours from the image sections; combining and averaging of extracted contours; and 3D reconstruction of the averaged contours. The generic models reconstructed from the averaged contours of six cadaver femora were validated by comparing their surface geometry on a point to point basis with that of the CT reconstructed reference models. The mean errors ranged from 0.99 to 2.5 mm and were in agreement with the qualitative assessment of the models. PMID- 17132617 TI - A generic 3-dimensional system to mimic trabecular bone surface adaptation. AB - The paper presents the structure optimizing system based on surface remodelling. The grounds for algorithm formulation are given by the bone remodelling phenomenon leading to optimization of trabecular network in the bone. The assumptions, algorithms and limitations of the own mesh generator Cosmoprojector are described. Unlike other approaches, the system is able to mimic real bone evolution including tissue consolidation and separation. The article presents a closed system consisting of finite element mesh generation, decision criteria for structure adaptation and finite element analysis in parallel environment. It also provides some computation results obtained by using specially designed software. PMID- 17132618 TI - A finite element model of the human left ventricular systole. AB - Local wall stress is the pivotal determinant of the heart muscle's systolic function. Under in vivo conditions, however, such stresses cannot be measured systematically and quantitatively. In contrast, imaging techniques based on magnetic resonance (MR) allow the determination of the deformation pattern of the left ventricle (LV) in vivo with high accuracy. The question arises to what extent deformation measurements are significant and might provide a possibility for future diagnostic purposes. The contractile forces cause deformation of LV myocardial tissue in terms of wall thickening, longitudinal shortening, twisting rotation and radial constriction. The myocardium is thereby understood to act as a densely interlaced mesh. Yet, whole cycle image sequences display a distribution of wall strains as function of space and time heralding a significant amount of inhomogeneity even under healthy conditions. We made similar observations previously by direct measurement of local contractile activity. The major reasons for these inhomogeneities derive from regional deviations of the ventricular walls from an ideal spheroidal shape along with marked disparities in focal fibre orientation. In response to a lack of diagnostic tools able to measure wall stress in clinical routine, this communication is aimed at an analysis and functional interpretation of the deformation pattern of an exemplary human heart at end-systole. To this end, the finite element (FE) method was used to simulate the three-dimensional deformations of the left ventricular myocardium due to contractile fibre forces at end-systole. The anisotropy associated with the fibre structure of the myocardial tissue was included in the form of a fibre orientation vector field which was reconstructed from the measured fibre trajectories in a post mortem human heart. Contraction was modelled by an additive second Piola-Kirchhoff active stress tensor. As a first conclusion, it became evident that longitudinal fibre forces, cross-fibre forces and shear along with systolic fibre rearrangement have to be taken into account for a useful modelling of systolic deformation. Second, a realistic geometry and fibre architecture lead to typical and substantially inhomogeneous deformation patterns as they are recorded in real hearts. We therefore, expect that the measurement of systolic deformation might provide useful diagnostic information. PMID- 17132620 TI - Proteomics identification of differentially expressed proteins associated with pollen germination and tube growth reveals characteristics of germinated Oryza sativa pollen. AB - Mature pollen from most plant species is metabolically quiescent; however, after pollination, it germinates quickly and gives rise to a pollen tube to transport sperms into the embryo sac. Because methods for collecting a large amount of in vitro germinated pollen grains for transcriptomics and proteomics studies from model plants of Arabidopsis and rice are not available, molecular information about the germination developmental process is lacking. Here we describe a method for obtaining a large quantity of in vitro germinating rice pollen for proteomics study. Two-dimensional electrophoresis of approximately 2300 protein spots revealed 186 that were differentially expressed in mature and germinated pollen. Most showed a changed level of expression, and only 66 appeared to be specific to developmental stages. Furthermore 160 differentially expressed protein spots were identified on mass spectrometry to match 120 diverse protein species. These proteins involve different cellular and metabolic processes with obvious functional skew toward wall metabolism, protein synthesis and degradation, cytoskeleton dynamics, and carbohydrate/energy metabolism. Wall metabolism related proteins are prominently featured in the differentially expressed proteins and the pollen proteome as compared with rice sporophytic proteomes. Our study also revealed multiple isoforms and differential expression patterns between isoforms of a protein. These results provide novel insights into pollen function specialization. PMID- 17132621 TI - Understanding the computational methodologies of systems biology. PMID- 17132622 TI - Representing, storing and accessing molecular interaction data: a review of models and tools. AB - One important aim within systems biology is to integrate disparate pieces of information, leading to discovery of higher-level knowledge about important functionality within living organisms. This makes standards for representation of data and technology for exchange and integration of data important key points for development within the area. In this article, we focus on the recent developments within the field. We compare the recent updates to the three standard representations for exchange of data SBML, PSI MI and BioPAX. In addition, we give an overview of available tools for these three standards and a discussion on how these developments support possibilities for data exchange and integration. PMID- 17132623 TI - The crystal structure and mutational binding analysis of the extracellular domain of the platelet-activating receptor CLEC-2. AB - The human C-type lectin-like molecule CLEC-2 is expressed on the surface of platelets and signaling through CLEC-2 causes platelet activation and aggregation. CLEC-2 is a receptor for the platelet-aggregating snake venom protein rhodocytin. It is also a newly identified co-receptor for human immunodeficiency virus type 1 (HIV-1). An endogenous ligand has not yet been identified. We have solved the crystal structure of the extracellular domain of CLEC-2 to 1.6-A resolution, and identified the key structural features involved in ligand binding. A semi-helical loop region and flanking residues dominate the surface that is available for ligand binding. The precise distribution of hydrophobic and electrostatic features in this loop will determine the nature of any endogenous ligand with which it can interact. Major ligand-induced conformational change in CLEC-2 is unlikely as its overall fold is compact and robust. However, ligand binding could induce a tilt of a 3-10 helical portion of the long loop region. Mutational analysis and surface plasmon resonance binding studies support these observations. This study provides a framework for understanding the effects of rhodocytin venom binding on CLEC-2 and for understanding the nature of likely endogenous ligands and will provide a basis for rational design of drugs to block ligand binding. PMID- 17132624 TI - Crystal structure of the tyrosine kinase domain of colony-stimulating factor-1 receptor (cFMS) in complex with two inhibitors. AB - The cFMS proto-oncogene encodes for the colony-stimulating factor-1 receptor, a receptor-tyrosine kinase responsible for the differentiation and maturation of certain macrophages. Upon binding its ligand colony-stimulating factor-1 cFMS autophosphorylates, dimerizes, and induces phosphorylation of downstream targets. We report the novel crystal structure of unphosphorylated cFMS in complex with two members of different classes of drug-like protein kinase inhibitors. cFMS exhibits a typical bi-lobal kinase fold, and its activation loop and DFG motif are found to be in the canonical inactive conformation. Both ATP competitive inhibitors are bound in the active site and demonstrate a binding mode similar to that of STI-571 bound to cABL. The DFG motif is prevented from switching into the catalytically competent conformation through interactions with the inhibitors. Activation of cFMS is also inhibited by the juxtamembrane domain, which interacts with residues of the active site and prevents formation of the activated kinase. Together the structures of cFMS provide further insight into the autoinhibition of receptor-tyrosine kinases via their respective juxtamembrane domains; additionally the binding mode of two novel classes of kinase inhibitors will guide the design of novel molecules targeting macrophage-related diseases. PMID- 17132625 TI - Protein engineering of the colony-stimulating factor-1 receptor kinase domain for structural studies. AB - A parallel approach to designing crystallization constructs for the c-FMS kinase domain was implemented, resulting in proteins suitable for structural studies. Sequence alignment and limited proteolysis were used to identify and eliminate unstructured and surface-exposed domains. A small library of chimeras was prepared in which the kinase insert domain of FMS was replaced with the kinase insert domain of previously crystallized receptor-tyrosine kinases. Characterization of the newly generated FMS constructs by enzymology and thermoshift assays demonstrated similar activities and compound binding to the FMS full-length cytoplasmic domain. Two chimeras were evaluated for crystallization in the presence and absence of a variety of ligands resulting in crystal structures, and leading to a successful structure-based drug design project for this important inflammation target. PMID- 17132626 TI - ATP activates a reactive oxygen species-dependent oxidative stress response and secretion of proinflammatory cytokines in macrophages. AB - Secretion of the proinflammatory cytokines, interleukin (IL)-1beta and IL-18, usually requires two signals. The first, due to microbial products such as lipopolysaccharide, initiates transcription of the cytokine genes and accumulation of the precursor proteins. Cleavage and secretion of the cytokines is mediated by caspase-1, in association with an inflammasome containing Nalp3, which can be activated by binding of extracellular ATP to purinergic receptors. We show that treatment of macrophages with ATP results in production of reactive oxygen species (ROS), which stimulate the phosphatidylinositol 3-kinase (PI3K) pathway and subsequent Akt and ERK1/2 activation. ROS exerts its effect through glutathionylation of PTEN (phosphatase and tensin homologue deleted from chromosome 10), whose inactivation would shift the equilibrium in favor of PI3K. ATP-dependent ROS production and PI3K activation also stimulate transcription of genes required for an oxidative stress response. In parallel, ATP-mediated ROS dependent PI3K is required for activation of caspase-1 and secretion of IL-1beta and IL-18. Thus, an increase in ROS levels in ATP-treated macrophages results in activation of a single pathway that promotes both adaptation to subsequent exposure to oxidants or inflammation, and processing and secretion of proinflammatory cytokines. PMID- 17132627 TI - Preservation of antimicrobial properties of complement peptide C3a, from invertebrates to humans. AB - The human anaphylatoxin peptide C3a, generated during complement activation, exerts antimicrobial effects. Phylogenetic analysis, sequence analyses, and structural modeling studies paired with antimicrobial assays of peptides from known C3a sequences showed that, in particular in vertebrate C3a, crucial structural determinants governing antimicrobial activity have been conserved during the evolution of C3a. Thus, regions of the ancient C3a from Carcinoscorpius rotundicauda as well as corresponding parts of human C3a exhibited helical structures upon binding to bacterial lipopolysaccharide permeabilized liposomes and were antimicrobial against gram-negative and gram positive bacteria. Human C3a and C4a (but not C5a) were antimicrobial, in concert with the separate evolutionary development of the chemotactic C5a. Thus, the results demonstrate that, notwithstanding a significant sequence variation, functional and structural constraints imposed on C3a during evolution have preserved critical properties governing antimicrobial activity. PMID- 17132628 TI - FOXO3a induces differentiation of Bcr-Abl-transformed cells through transcriptional down-regulation of Id1. AB - Leukemic transformation often requires activation of protein kinase B (PKB/c-Akt) and is characterized by increased proliferation, decreased apoptosis, and a differentiation block. PKB phosphorylates and inactivates members of the FOXO subfamily of Forkhead transcription factors. It has been suggested that hyperactivation of PKB maintains the leukemic phenotype through actively repressing FOXO-mediated regulation of specific genes. We have found expression of the transcriptional repressor Id1 (inhibitor of DNA binding 1) to be abrogated by FOXO3a activation. Inhibition of PKB activation or growth factor deprivation also resulted in strong down-regulation of Id1 promoter activity, Id1 mRNA, and protein expression. Id1 is highly expressed in Bcr-Abl-transformed K562 cells, correlating with high PKB activation and FOXO3a phosphorylation. Inhibition of Bcr-Abl by the chemical inhibitor STI571 resulted in activation of FOXO3a and down-regulation of Id1 expression. By performing chromatin immunoprecipitation assays and promoter-mutation analysis, we demonstrate that FOXO3a acts as a transcriptional repressor by directly binding to the Id1 promoter. STI571 treatment, or expression of constitutively active FOXO3a, resulted in erythroid differentiation of K562 cells, which was inhibited by ectopic expression of Id1. Taken together our data strongly suggest that high expression of Id1, through PKB mediated inhibition of FOXO3a, is critical for maintenance of the leukemic phenotype. PMID- 17132629 TI - Evolutionary processes during the formation of the plant-specific Dof transcription factor family. AB - We found 19 putative genes for plant-specific Dof transcription factors in the moss Physcomitrella patens and one Dof gene in the green alga Chlamydomonas reinhardtii, but no identifiable Dof gene in the red alga Cyanidioschyzon merolae and the diatom Thalassiosira pseudonana, suggesting that the origin of the Dof transcription factors pre-dates the divergence of the green algae and the ancestors of terrestrial plants. The phylogenetic analyses contended that the Dof family in angiosperms formed through a series of evolutionary processes, including intensive duplications of a specific ancestral gene after the divergence of the moss and the angiosperm lineages. PMID- 17132630 TI - Comparative overviews of clock-associated genes of Arabidopsis thaliana and Oryza sativa. AB - In higher plants, circadian rhythms are highly relevant to a wide range of biological processes. To such circadian rhythms, the clock (oscillator) is central, and recent intensive studies on the model higher plant Arabidopsis thaliana have begun to shed light on the molecular mechanisms underlying the functions of the central clock. Such representative clock-associated genes of A. thaliana are the homologous CCA1 and LHY genes, and five PRR genes that belong to a small family of pseudo-response regulators including TOC1. Others are GI, ZTL, ELF3, ELF4, LUX/PCL1, etc. In this context, a simple question arose as to whether or not the molecular picture of the model Arabidopsis clock is conserved in other higher plants. Here we made an effort to answer the question with special reference to Oryza sativa, providing experimental evidence that this model monocot also has a set of highly conserved clock-associated genes, such as those designated as OsCCA1, OsPRR-series including OsTOC1/OsPRR1, OsZTLs, OsPCL1 as well as OsGI. These results will provide us with insight into the general roles of plant circadian clocks, such as those for the photoperiodic control of flowering time that has a strong impact on the reproduction and yield in many higher plants. PMID- 17132631 TI - Identification of a WRKY protein as a transcriptional regulator of benzylisoquinoline alkaloid biosynthesis in Coptis japonica. AB - Selected cultured Coptis japonica cells produce a large amount of the benzylisoquinoline alkaloid berberine. Previous studies have suggested that berberine productivity is controlled at the transcript level of biosynthetic genes. We have identified a regulator of transcription in berberine biosynthesis using functional genomics with a transient RNA interference (RNAi) and overexpression of the candidate gene. The 24 primary candidate clones were selected from 1,014 expressed sequence tags (ESTs) that were obtained from a C. japonica cell line producing high levels of berberine. Further characterization of the expression profiles of these ESTs suggested that five ESTs would be good candidates as regulators of berberine production. A newly developed transient RNAi system with C. japonica protoplasts indicated that double-stranded RNA of an EST clone significantly reduced the level of transcripts of 3'-hydroxy N methylcoclaurine 4'-O-methyltransferase. Sequence analysis showed that this EST encoded a group-II WRKY, and we named it CjWRKY1. When the effects of double stranded RNA of the CjWRKY1 gene were examined in detail, a marked reduction in the transcripts of all genes involved in berberine biosynthesis was detected, whereas little effect was found in the transcript levels of glyceraldehyde-3 phosphate dehydrogenase (GAPDH) and chorismate mutase (CM) that are associated with primary metabolism. Ectopic expression of CjWRKY1 cDNA in C. japonica protoplasts clearly increased the level of transcripts of all berberine biosynthetic genes examined compared with control treatment, whereas the levels of GAPDH and CM were not affected. The functional role of CjWRKY1 as a specific and comprehensive regulator of berberine biosynthesis is discussed. PMID- 17132632 TI - Type-B ARR transcription factors, ARR10 and ARR12, are implicated in cytokinin mediated regulation of protoxylem differentiation in roots of Arabidopsis thaliana. AB - In the phosphorelay-mediated cytokinin signal transduction of Arabidopsis thaliana, certain members of the type-B authentic response regulator (ARR) family are implicated in the regulatory networks that are primarily propagated by the cytokinin-receptors [authentic histidine kinases (AHKs)]. Clarification of the involvement of each type-B ARR transcription factor in cytokinin-responsive phenomena is still at a very early stage. Here we analyzed the redundant function of two type-B ARR genes, ARR10 and ARR12, by constructing an arr10/arr12 double mutant. The resulting mutant plants showed stronger phenotypes with special reference to the cytokinin action in roots (e.g. inhibition of root elongation, green callus formation from root explants) than those for each single mutant, suggesting that ARR10 and ARR12 redundantly play an important role in the cytokinin signaling in roots. This idea was further supported by results from root-specific microarray analyses with the double mutant plant. We also showed that ARR10 and ARR12 are involved in the AHK-dependent signaling pathway that negatively regulates protoxylem specification in root vascular tissues. When the double mutant is combined with an arr1 allele, the resultant arr1/arr10/arr12 triple mutant showed phenotypes displaying a very poor growth, quite similar to those of the wooden leg (wol) mutant that virtually lacks cytokinin receptor activities in plants. In this triple arr mutant, the specification of root vascular tissues is also affected as severely as in wol. Taken together, we propose that ARR10 and ARR12, together with ARR1, redundantly play pivotal roles in the AHK-dependent phosphorelay signaling in response to cytokinin in roots. PMID- 17132633 TI - Co-regulation of brassinosteroid biosynthesis-related genes during xylem cell differentiation. AB - To understand the regulatory mechanisms of brassinosteroid (BR) biosynthesis in specific plant developmental processes, we first investigated the accumulation profiles of BRs and sterols in xylem differentiation in a Zinnia culture. The amounts of many substances in the late C28 sterol biosynthetic pathway to campesterol (CR), such as episterol and 24-methylenecholesterol, as well as those in the BR-specific biosynthetic pathway from CR to brassinolide (BL), were elevated in close association with tracheary element differentiation. Among them, 6-deoxotyphasterol (6-deoxoTY) accumulated to unusually high levels within cells cultured in tracheary element-inductive medium, while castasterone (CS) was not elevated either within or outside cells. To identify the molecular basis of this co-up-regulation of BRs and C28 sterols, we isolated Zinnia genes for the key enzymes of BR biosynthesis, ZeSTE1, ZeDIM, ZeDWF4, ZeCPD1 and ZeCPD2. RNA gel blot analysis of these genes indicated a coordinated increase in transcripts for ZeSTE1, ZeDIM, ZeDWF4 and ZeCPD1, and a tracheary element differentiation specific increase in transcripts for ZeDWF4 and ZeCPD1. In situ hybridization experiments of ZeDWF4 and ZeCPD1 mRNAs revealed their preferential accumulation in procambium cells, immature xylem cells and xylem parenchyma cells. These results suggest that BR biosynthesis during tracheary element differentiation may be regulated by the coordinated regulation of broad sterol biosynthesis and specific regulation of BR biosynthesis, which occurs in part by elevated transcript levels of genes encoding BR biosynthetic enzymes, specifically ZeDWF4 and ZeCPD1. These data provide new insights into the regulation of BR biosynthesis and BR signaling during plant development. PMID- 17132634 TI - Magnesium enhances aluminum-induced citrate secretion in rice bean roots (Vigna umbellata) by restoring plasma membrane H+-ATPase activity. AB - We demonstrated that magnesium (Mg) can alleviate aluminum (Al) toxicity in rice bean [Vigna umbellata (Thunb.) Ohwi & Ohashi] more effectively than is expected from a non-specific cation response. Micromolar concentrations of Mg alleviated the inhibition of root growth by Al but not by lanthanum, and neither strontium nor barium at the micromolar level alleviates Al toxicity. Aluminum also induced citrate efflux from rice bean roots, and this response was stimulated by inclusion of 10 microM Mg in the treatment solution. The increase in the Al induced citrate efflux by Mg paralleled the improvement in root growth, suggesting that the ameliorative effect of Mg might be related to greater citrate efflux. Vanadate (an effective H+-ATPase inhibitor) decreased the Al-induced citrate efflux, while addition of Mg partly restored the efflux. Mg addition also increased the activity of Al-reduced plasma membrane H+-ATPase, as well as helping to maintain the Mg and calcium contents in root apices. We propose that the addition of Mg to the toxic Al treatment helps maintain the tissue Mg content and the activity of the plasma membrane H+-ATPase. These changes enhanced the Al dependent efflux of citrate which provided extra protection from Al stress. PMID- 17132635 TI - Age at disability milestones in multiple sclerosis and history of multiple sclerosis: a unifying concept. PMID- 17132638 TI - Deep brain stimulation in Parkinson's disease can mimic the 300 Hz subthalamic rhythm. PMID- 17132640 TI - Independent patterns of damage within magno-, parvo- and koniocellular pathways in Parkinson's disease (Silva MF et al. Brain 2005; 128: 2260-2271). PMID- 17132643 TI - Axonal protection in multiple sclerosis--a particular need during remyelination? PMID- 17132644 TI - Predictive performance of 'Servin's formula' during BIS-guided propofol remifentanil target-controlled infusion in morbidly obese patients. AB - BACKGROUND: The aim of this study was to assess the predictive performance of 'Servin's formula' for bispectral index (BIS)-guided propofol-remifentanil target controlled infusion (TCI) in morbidly obese patients. METHODS: Twenty patients (ASA physical status II-III, age 32-64 yr) undergoing bilio-intestinal bypass surgery, were recruited. Anaesthesia was induced by using a TCI of propofol with an initial target plasma concentration of 6 microg ml(-1), then adapted to maintain stable BIS values ranging between 40 and 50. A TCI of remifentanil was added to achieve pain control and haemodynamic stability. For propofol, weight was corrected as suggested by Servin and colleagues. With ideal body weight (IBW) corrected according to formula suggested by Lemmens and colleagues. For remifentanil, weight was corrected according to IBW. Arterial blood samples for the determination of blood propofol concentrations were collected at different surgical times. The predictive performance of propofol TCI was evaluated by examining performance accuracy. RESULTS: Median prediction error and median absolute prediction error were -32.6% (range -53.4%; -2.5%) and 33.1% (10.8%; 53.4%), respectively. Wobble median value was 5.9% (2.5%; 25.2%) while divergence median value was -1.5% h(-1) (-7.7; 33.8% h(-1)). CONCLUSION: Significant bias between predicted and measured plasma propofol concentrations was found while the low wobble values suggest that propofol TCI system is able to maintain stable drug concentrations over time. As already suggested before, a computer simulation confirmed that the TCI system performance could be significantly improved when total body weight is used. PMID- 17132645 TI - Improving tomorrow's health care with today's tools: a conversation with Larry C. Glasscock. Interview by John K. Iglehart. AB - The chairman and CEO of WellPoint Inc. provides some factual background on one of the nation's largest health insurers, which covers 34.2 million lives. Its own employees are offered the opportunity to join a consumer-directed health plan, which gives the company some real-life data on which to base its strategies in the marketplace. Glasscock believes that future reform efforts in his company and at the national level must focus on improving quality and affordability, reducing the number of uninsured Americans, and improving the health of those insured by WellPoint and everyone else. PMID- 17132646 TI - Albuminuria as risk factor for initiation and progression of carotid atherosclerosis in non-diabetic persons: the Tromso Study. AB - AIMS: High levels of microalbuminuria have been associated with severe atherosclerosis. In this prospective, population-based study, we examined whether urinary albumin-to-creatinine-ratios (ACR) in the lower range were associated with the initiation and progression of atherosclerosis. METHODS AND RESULTS: Carotid ultrasonography and measurements of ACR, fibrinogen, monocytes, white cell count, and well-established cardiovascular risk factors were performed in 4037 non-diabetic subjects, 2203 without, and 1834 with pre-existing plaques at baseline. After 7 years new ultrasound measurements were performed. In subjects without pre-existing plaques, 884 had developed at least one plaque during follow up. Baseline ACR was significantly related to the area of the novel plaques (P for linear trend = 0.009 over the baseline ACR quartiles, after multiple adjustments). The relationship with ACR was clearly modified by fibrinogen (P = 0.001, for the interaction ACR x fibrinogen). Subjects with high levels of both ACR and fibrinogen developed plaques with the largest area. In subjects with pre existing plaques, ACR was related to plaque-progression (P for linear trend = 0.026, after multiple adjustments). In these individuals, the interaction between fibrinogen and ACR on plaque-growth appeared only in those with minimal atherosclerosis at baseline. CONCLUSION: ACR is positively related to plaque initiation and plaque-growth. This relationship is substantially modified by fibrinogen in previously plaque-free subjects. PMID- 17132647 TI - Carbohydrate nutrition, glycaemic load, and plasma lipids: the Insulin Resistance Atherosclerosis Study. AB - AIMS: We evaluated the relationship of carbohydrate nutrition and selected food groups with lipids using data from the Insulin Resistance Atherosclerosis Study (IRAS Exam I, 1992-1994). METHODS AND RESULTS: A total of 1026 middle-aged adults with normal or impaired glucose tolerance had complete data on fasting lipids and usual dietary intake from an interviewer-administered, validated food frequency questionnaire. Published glycaemic index (GI) values were assigned to food items and average dietary GI and glycaemic load (GL) were calculated per participant. Intake of carbohydrates differed by gender, men consuming more absolute digestible carbohydrates with higher GI and GL than women. In multivariate models adjusting for energy intake, in men, GL and carbohydrates were associated positively with total and LDL cholesterol, and inversely with HDL. In women, associations were limited to triglycerides. We estimated that a 100 g higher intake in GL or carbohydrates was associated with a 7-8 mg/dL higher total or LDL cholesterol level in men, and a 13-17 mg/dL higher triglyceride level in women. In the combined sample, GL and carbohydrates were consistently associated with all lipid levels and GI was inversely associated with HDL cholesterol. CONCLUSION: Our findings underscore the importance of carbohydrate nutrition for plasma lipids. PMID- 17132648 TI - Renal function and cardiovascular mortality in elderly men: the role of inflammatory, procoagulant, and endothelial biomarkers. AB - AIMS: To assess the extent to which inflammatory, procoagulant, and endothelial biomarkers modify the relationship between diminished renal function and cardiovascular mortality. METHODS AND RESULTS: Prospective study of 4029 men aged 60-79 years followed up for a mean period of 6 years, during which 304 cardiovascular deaths occurred. Predicted estimated glomerular filtration rate (eGFR) was used as a measure of renal function. Reduced eGFR was associated with increased prevalence of established cardiovascular risk factors [cardiovascular disease, diabetes, hypertension, left ventricular (LV) hypertrophy, and dyslipidaemia] and higher levels of inflammatory markers [interleukin 6 (IL-6), C reactive protein], endothelial markers [von Willebrand factor (vWF) and tissue plasminogen activator], activated coagulation markers (fibrin D-dimer), and blood viscosity. Cardiovascular mortality risk increased with decreasing levels of eGFR, particularly among men with eGFR <60 mL/min per 1.73 m(2) even after adjustment for established risk factors (adjusted RR 1.49, 95% CI 1.10, 2.03; <60 vs. > or =70 mL/min per 1.73 m(2)). The association was attenuated after further adjustment for vWF, D-dimer, and IL-6 (adjusted RR 1.34, 95% CI 0.98-1.82). CONCLUSION: Mild-to-moderate renal insufficiency is associated with significantly increased cardiovascular mortality in elderly men, which is partly explained by the increased prevalence of established risk factors, markers of coagulation, endothelium, and inflammation. PMID- 17132649 TI - Impact of C-reactive protein and fibrinogen on cardiovascular prognosis in patients with stable angina pectoris: the AtheroGene study. AB - AIMS: C-reactive protein and fibrinogen have been extensively studied and shown to be predictive for a first cardiovascular event in healthy individuals. We evaluated the potential clinical use of C-reactive protein and fibrinogen in patients already suffering from coronary artery disease (CAD). METHODS AND RESULTS: In a substudy of the prospective AtheroGene registry, we assessed in 1806 patients with documented CAD and stable angina pectoris, the risk of cardiovascular death and non-fatal myocardial infarction (n=183) over a median follow-up of 3.5 (maximum 7.7) years according to baseline levels of C-reactive protein and fibrinogen. C-reactive protein and fibrinogen were associated with future cardiovascular events, such as an increment in one standard deviation of C reactive protein is associated with a 1.15-fold (95% CI 1.05-1.27, P=0.002) increase, an increment of one standard deviation of fibrinogen with a 1.27-fold (95% CI 1.12-1.43, P<0.0005) increase in hazard risk in the models adjusted for age and sex. Adjustment for traditional risk factors and clinical confounders did not significantly attenuate this relationship. In a comparison of a basic model (traditional risk factors; AUC=0.68) with models additionally including either C reactive protein (AUC=0.69) or fibrinogen (AUC=0.70), only little additional predictive information over that obtained from assessment of traditional risk factors was obtained. CONCLUSION: In patients with documented CAD, C-reactive protein and fibrinogen were predictive for future cardiovascular risk, but did not provide further information on top of that obtained from models including traditional risk factors. Our data emphasize the clinical importance of traditional risk factors in patients with CAD. PMID- 17132650 TI - Pretreatment with intracoronary adenosine reduces the incidence of myonecrosis after non-urgent percutaneous coronary intervention: a prospective randomized study. AB - AIMS: We sought to investigate the effect of adenosine pretreatment on the incidence of myonecrosis after non-urgent percutaneous coronary intervention (PCI). METHODS AND RESULTS: This was a prospective, randomized, open-label study. Patients who were scheduled for non-urgent PCI in de novo native coronary arteries were eligible. All patients were pretreated with aspirin and clopidogrel. Myonecrosis was measured by creatine kinase-myocardial band (CK-MB) elevation after PCI. A total of 62 patients were randomized into the adenosine (n = 31) or standard (n = 31) group. The adenosine group received 50 microg adenosine bolus before wiring of each lesion, whereas the standard group did not. Post-PCI myonecrosis occurred more frequently in the standard group (39 vs. 13%, OR 0.23, 95% CI 0.05-0.95, P = 0.020). After adjustment for drug-eluting stent implantation, multi-vessel stenting, and elevated baseline troponin, the OR was 0.19 (95% CI 0.05-0.72, P = 0.017). The median peak values of CK-MB in the adenosine and standard groups were 2 and 4 microg/L, respectively (P = 0.033). The adjusted difference was 1.95 microg/L (95% CI 0.13-3.77, P = 0.037). The incidences of myocardial infarction (>3 x CK-MB) were 6 and 16% in the adenosine and standard groups, respectively (OR 0.36; 95% CI 0.03-2.46, P = 0.229). CONCLUSION: Pretreatment with 50 microg of adenosine decreases the incidence of myonecrosis after non-urgent PCI compared with that without pretreatment. PMID- 17132651 TI - Molecular signature of recovery following combination left ventricular assist device (LVAD) support and pharmacologic therapy. AB - AIMS: A novel combination therapy consisting of a left ventricular assist device (LVAD) combined with pharmacologic therapy including the selective beta(2) agonist, clenbuterol, has shown promise in restoring ventricular function in patients with heart failure. The aim of this study was to identify common genes and signalling pathways whose expression was associated with reversal of heart failure and restoration of ventricular function. METHODS AND RESULTS: Microarray analysis was performed on six paired human heart samples harvested at the time of LVAD implant and at the time of LVAD explant for recovery of ventricular function (post). Follow-up data shows that the improvements in ventricular function have been maintained for an average of 3.8 years post-explant. Analysis of the gene expression data revealed: (i) a significant association of integrin pathway signalling with recovery and (ii) the identification of several novel targets including, EPAC2, in the well-described cAMP pathway whose expression was down regulated with recovery, and was associated with improvements in cardiac contractility, metabolism, and function. CONCLUSION: This data set represents the first description of signalling pathways associated with the functional recovery of end-stage human heart failure and the identification of new targets in the human heart that are modified by this combination therapy. PMID- 17132652 TI - Infarct-related left ventricular diverticulum. PMID- 17132653 TI - A clinical survival score predicts the likelihood to benefit from preoperative thallium scanning and coronary revascularization before major vascular surgery. AB - AIMS: Guidelines advocate selective non-invasive testing before major non-cardiac surgery, yet data defining who may benefit from such tests is lacking. We aimed to find the predictors that define patients who are most likely to benefit from preoperative cardiac testing and coronary revascularization (CR). METHODS AND RESULTS: In 624 consecutive major vascular surgery patients, the preoperative thallium scanning (PTS) results and subsequent CRs were correlated with long-term (3-15 years) survival. Of all patients, 510 (80.6%) had PTS, 154 (24.7%) had moderate-severe ischaemia on PTS, and 96 (15.4%) underwent CR. Seven predictors: age >or=65, diabetes, cerebrovascular disease, ischaemic heart disease, congestive heart failure, ST-depression on preoperative ECG, and renal insufficiency, independently determined long-term survival. A long-term survival score (LTSS) comprised of these predictors, divided all patients into low, intermediate, and high-risk groups (0-1, 2-3, >or=4 predictors, respectively) with a 5-year survival of 83 +/- 2%, 60 +/- 3%, and 34 +/- 6%, respectively. Compared with low-risk patients, intermediate and high-risk patients had worse survival [HR (CI) = 2.6 (2.0-3.4) and 5.9 (4.1-8.9), respectively, P < 0.001]. Yet, only the intermediate-risk group had better long-term survival following preoperative CR [HR = 0.48 (0.31-0.75), P = 0.001]. The low-risk groups' favourable survival and high-risk groups' poor survival were not significantly affected by CR. CONCLUSION: Intermediate-risk patients (LTSS 2-3) are most likely to have a long-term survival benefit from PTS and CR. PMID- 17132654 TI - Outcomes after normal dobutamine stress echocardiography and predictors of adverse events: long-term follow-up of 3014 patients. AB - AIMS: Normal exercise echocardiography predicts a good prognosis. Dobutamine stress echocardiography (DSE) is generally reserved for patients with comorbidities which preclude exercise testing. We evaluated predictors of adverse events after normal DSE. METHODS AND RESULTS: We studied 3014 patients (1200 males, 68+/-12 years) with normal DSE, defined as the absence of wall motion abnormality at rest or with stress. During median follow-up of 6.3 years, all cause mortality and cardiac events, defined as myocardial infarction and coronary revascularization, occurred in 920 (31%) and 231 (7.7%) patients, respectively. Survival and cardiac event-free probabilities were 95 and 98% at 1 year, 78 and 93% at 5 years, and 56 and 89% at 10 years, respectively. Age, diabetes mellitus, and failure to achieve 85% age-predicted maximal heart rate were independent predictors of mortality and cardiac events. Patients with all three of these characteristics had a 13% probability of cardiac events within the first year and higher risk throughout follow-up. CONCLUSION: Prognosis after normal DSE is not necessarily benign, but depends on patient and stress test characteristics. Careful evaluation, using clinical and stress data, is required to identify patients with normal DSE who are at increased risk of adverse outcomes during long-term follow-up. PMID- 17132655 TI - IMBA Professional Plus: a flexible approach to internal dosimetry. AB - IMBA (Integrated Modules for Bioassay Analysis) is a suite of software modules that implement the current ICRP biokinetic and dosimetric models for estimation of intakes and doses. The IMBA modules have gone through extensive quality assurance, and are now used for routine formal dose assessment by Approved Dosimetry Services throughout the UK. HPA has continued to develop the IMBA modules. In addition, several projects, sponsored by organisations both in the USA and in Canada, have resulted in the development of customised user-friendly interfaces (IMBA Expert 'editions'). These enable users not only to use the standard ICRP models, but also to change many of the parameter values from ICRP defaults, and to apply sophisticated data handling techniques to internal dose calculations. These include: fitting measurement data with the maximum likelihood method; using multiple chronic and acute intakes; and dealing with different data types, such as urine, faces and whole body simultaneously. These interfaces were improved further as a result of user-feedback, and a general 'off-the-shelf' product, IMBA Professional, was developed and made available in January 2004. A new version, IMBA Professional Plus, was released in April 2005, which is both faster and more powerful than previous software. The aim of this paper is to describe the capabilities of IMBA Professional Plus, and the mathematical methods used. PMID- 17132656 TI - Development of a calculation method for estimating specific energy distribution in complex radiation fields. AB - Estimation of the specific energy distribution in a human body exposed to complex radiation fields is of great importance in the planning of long-term space missions and heavy ion cancer therapies. With the aim of developing a tool for this estimation, the specific energy distributions in liquid water around the tracks of several HZE particles with energies up to 100 GeV n(-1) were calculated by performing track structure simulation with the Monte Carlo technique. In the simulation, the targets were assumed to be spherical sites with diameters from 1 nm to 1 microm. An analytical function to reproduce the simulation results was developed in order to predict the distributions of all kinds of heavy ions over a wide energy range. The incorporation of this function into the Particle and Heavy Ion Transport code System (PHITS) enables us to calculate the specific energy distributions in complex radiation fields in a short computational time. PMID- 17132657 TI - DNA damage induced by the direct effect of He ion particles. AB - We present here evidence showing that the yields of DNA lesions induced by He(2+) ions strongly depend on Linear energy transfer (LET). In this study, hydrated plasmid DNA was irradiated with He(2+) ions with LET values of 19, 63 and 95 keVmicrom(-1). The yields of prompt single-strand breaks (SSBs) are very similar at the varying LET values, whereas the yields of prompt double-strand breaks (DSBs) increase with increasing LET. Further, base lesions were revealed as additional strand breaks by post-irradiation treatment of the DNA with endonuclease III (Nth) and formamidopyrimidine-DNA glycosylase (Fpg). The reduction in the yield of these enzymatically induced SSBs and DSBs becomes significant as the LET increases. These results suggest that the clustering of DNA lesions becomes more probable in regions of high LET. PMID- 17132658 TI - Radiation quality of tritium. AB - Tritium occurs from both natural and manufactured processes. In the environment, tritium can exist in the form of tritiated water (HTO) and in an organic form known as organically-bound tritium (OBT). Although, the concentrations of environmental OBT are relatively low, there is concern that current risk factors may underestimate the risk from OBT. Because tritium poses an internal hazard at cellular levels, microdosimetric techniques provide suitable tools for the study of radiation quality of tritium. In this study, microdosimetric simulations are performed for tritium uniformly distributed in a medium, and for tritium bound to biologically critical sites of dimensions from 10 nm to 2 microm. Results of local energy density are different for these two cases in microscopic regions. Based on the spatial distribution of energy deposition, dose mean lineal energies are calculated for tritium in forms of HTO and OBT. The dose mean lineal energies of OBT are about a factor of 1.7 higher than those of HTO in a wide range of target dimensions of biological interest. The results are consistent with radiobiological findings that OBT is about twice as effective as that of HTO. PMID- 17132659 TI - Monte Carlo modelling of energy deposition in trabecular bone. AB - This study includes the design and testing of a program that creates quadric based geometric models of the trabecular region, designed specifically for use with the 2005 version of the Monte Carlo radiation transport code PENELOPE. Our model was tested, by comparison with published data, in two aspects: the distributions of path lengths throughout the geometry and absorbed fraction values from the monoenergetic emission of electrons from within our geometry. In both comparisons, our results show a close agreement with published methods. PMID- 17132660 TI - A comparative review of charged particle microbeam facilities. AB - At the low doses (and low dose rates) relevant to environmental and occupational radiation exposure (0-50 mSv), which are of practical concern for radiation protection, very few cells in the human organism experience more than one traversal by densely ionising particles in their lifetime, the intervals between the tracks, if any, typically being months or years. The biological effects of exactly one particle are not well known and cannot be simulated in vitro by conventional broad-beam exposures, due to the random Poisson distribution of tracks. Charged particle microbeam facilities are a unique tool that allows targeting of single cells and analysis of the induced damage on a cell-by-cell basis. In the past few years, many charged particle microbeam facilities for radiobiology have come into operation or are under development worldwide. Different experimental designs have been adopted at various laboratories regarding the achievement of micrometre (or sub-micrometre) ion beam size, by mechanical collimation or focusing, particle detection, and cell recognition and positioning systems. The different approaches are reviewed and discussed in this paper. PMID- 17132661 TI - DNA-repair protein distribution along the tracks of energetic ions. AB - A simple model of homogenous chromatin distribution in HeLa-cell nuclei suggests that the track of an energetic ion hits 30 nm chromatin fibers with a mean distance of 0.55 mum. To test this assumption, living HeLa-cells were irradiated at the irradiation setup of the ion microprobe SNAKE using the ion beams provided by the Munich 14 MV tandem accelerator. After irradiation, the distribution of 53BP1 protein foci was studied by immunofluorescence. The observed 53BP1 distribution along the tracks of 29 MeV (7)Li ions and 24 MeV (12)C ions differed significantly from the expectations resulting from the simple chromatin model, suggesting that the biological track structure is determined by cell nuclear architecture with higher order organisation of chromatin. PMID- 17132662 TI - Alpha-particle radiobiological experiments using thin CR-39 detectors. AB - The present paper studied the feasibility of applying comet assay to evaluate the DNA damage in individual HeLa cervix cancer cells after alpha-particle irradiation. We prepared thin CR-39 detectors (<20 microm) as cell-culture substrates, with UV irradiation to shorten the track formation time. After irradiation of the HeLa cells by alpha particles, the tracks on the underside of the CR-39 detector were developed by chemical etching in (while floating on) a 14 N KOH solution at 37 degrees C. Comet assay was then applied. Diffusion of DNA out of the cells could be generally observed from the images of stained DNA. The alpha-particle tracks corresponding to the comets developed on the underside of the CR-39 detectors could also be observed by just changing the focal plane of the confocal microscope. PMID- 17132663 TI - A comparison of cellular irradiation techniques with alpha particles using the Geant4 Monte Carlo simulation toolkit. AB - A comparison of three cellular irradiation techniques using the Monte Carlo simulation toolkit Geant4 is presented in this paper. They involve electrodeposited source of alpha particle-emitting radionuclides, random classical alpha beam irradiation and single cell targeted irradiation using a focused alpha microbeam line. The simulation allows the calculation of hit distributions among the cellular population as well as the absorbed dose for two typical cellular geometries. PMID- 17132664 TI - Yields of strand breaks and base lesions induced by soft X-rays in plasmid DNA. AB - The yields of soft-X-ray-induced DNA damages have been measured by using closed circular plasmid DNA. Several DNA solutions with three kinds of radical scavenger capacity and also fully hydrated DNA samples were irradiated to compare the contribution by indirect reaction of diffusible water radicals, such as OH*, with those by direct action of secondary electrons. The yields of prompt single- (SSBs) and double-strand breaks (DSBs) decrease with increasing scavenging capacity. The SSB yields for soft X-rays are approximately midway those between gamma-ray and ultrasoft X-ray data previously reported. Heat labile sites are observed only in the low scavenger condition. The yields of the base lesions revealed by post irradiation treatment with base excision repair enzymes showed a similar value for Nth and Fpg protein except in the hydrated sample. These results indicate that the direct effect of soft X-rays induces the damages with different efficiency from those by indirect effect. PMID- 17132665 TI - Calculation of DNA strand breakage by neutralisation effect after 125I decays in a synthetic oligodeoxynucleotide using charge transfer theory. AB - The decay of the radioisotope (125)I into (125)Te is typically followed by the emission of two groups of approximately 10 electrons each via Auger processes. In deoxyribonucleic acid (DNA) with (125)I incorporated, these electrons produce various types of damage to DNA, e.g. single strand breaks (SSBs) and double strand breaks (DSBs) through direct actions of physical tracks, or indirect actions of radicals produced in water. Among the direct actions one should consider not only the excitation and ionisation of DNA by Auger electrons, but also the neutralisation of highly charged (125 m)Te ions with electrons from neighbouring molecules. Comparison between experiment and simulation done recently revealed that without including neutralisation effect the simulated yield of SSBs was 50% less than the measured result. In the present work a calculation of DNA strand breakage by the neutralization effect in a 41-mer synthetic oligodeoxynucleotide (oligoDNA) model was done using the charge transfer theory. Calculation based on transfer rate using the newly evaluated electronic coupling of DNA bases showed that the positive charge (hole) transfer rate is of the order of magnitudes of several 10(13) s(-1), implying that a charge higher than 10 units might not build on a (125 m)Te atom. The potential energy accumulated on the decay base is transferred to bases along the DNA chain nearby and destroys those bases and ionises the sugar-phosphate group, leading a DNA SSB with a frequency of 0.2% per eV in average. PMID- 17132666 TI - Charge transfer and ionisation by intermediate-energy heavy ions. AB - The use of heavy ion beams for microbeam studies of mammalian cell response leads to a need to better understand interaction cross sections for collisions of heavy ions with tissue constituents. For ion energies of a few MeV u(-1) or less, ions capture electrons from the media in which they travel and undergo subsequent interactions as partially 'dressed' ions. For example, 16 MeV fluorine ions have an equilibrium charge of 7(+), 32 MeV sulphur ions have an equilibrium charge of approximately 11(+), and as the ion energies decrease the equilibrium charge decreases dramatically. Data for interactions of partially dressed ions are extremely rare, making it difficult to estimate microscopic patterns of energy deposition leading to damage to cellular components. Such estimates, normally obtained by Monte Carlo track structure simulations, require a comprehensive database of differential and total ionisation cross sections as well as charge transfer cross sections. To provide information for track simulation, measurement of total ionisation cross sections have been initiated at East Carolina University using the recoil ion time-of-flight method that also yields cross sections for multiple ionisation processes and charge transfer cross sections; multiple ionisation is prevalent for heavy ion interactions. In addition, measurements of differential ionisation cross sections needed for Monte Carlo simulation of detailed event-by-event particle tracks are under way. Differential, total and multiple ionisation cross sections and electron capture and loss cross sections measured for C(+) ions with energies of 100 and 200 keV u(-1) are described. PMID- 17132667 TI - CFD as a tool in risk assessment of inhaled radon progenies. AB - During the last decade, computational fluid dynamics techniques proved to be a powerful tool in the modelling of biological processes and the design of biomedical devices. In this work, a computational fluid dynamics method was applied to model the transport of inhaled air and radioactive particles within the human respiratory tract. A finite volume numerical approach was used to compute the flow field characteristics and particle trajectories in the lumen of the first five airway generations of the human tracheobronchial tree, leading to the right upper lobe. The computations were performed for breathing and exposure conditions characteristic of uranium mines and homes. Primary radon daughter deposition patterns and energy distributions were computed, exhibiting highly inhomogeneous particle and energy deposition patterns. The results of the present modelling effort can serve as input data in lung cancer risk analysis. PMID- 17132668 TI - Type testing of an extremity finger stall dosemeter based on Harshaw TLD EXTRAD technology. AB - A new type of extremity dosemeter, which incorporates the Harshaw TLD EXTRAD dosemeter element into a PVC finger stall, has been developed. The dosemeter uses high-sensitivity lithium fluoride, (7)LiF:Mg,Cu,P (TLD-700H) in a thin 7 mg cm( 2) layer, with alternative coverings of PVC at 10 mg cm(-2) and aluminised polyester at 3.2 mg cm(-2). Results are presented of the type testing of both versions of the finger stall dosemeter against published standards. PMID- 17132669 TI - Proximity functions for electrons from 100 eV to 10 MeV. AB - The objective of this study is to provide a set of proximity functions for electrons from 100 eV to 10 MeV. Numerical results of differential proximity functions are given graphically. The complete data set is available electronically upon request from the authors. The results can serve as a convenient database for anyone performing microdosimetric calculations in radiation fields of electrons. For mixed fields of electrons, the proximity functions can easily be derived from the proximity functions of monoenergetic electrons presented here. PMID- 17132670 TI - Keeping up with the neighbours--measuring the bystander response. AB - Ionising radiation can induce responses within non-exposed neighbouring (bystander) cells, which potentially have important implications on the estimates of risk at environmentally relevant doses. Using human skin fibroblasts (AG1522), a range of methods were used to investigate the nature of the signal(s) arising from the exposed cells. The signal(s) can be transmitted by direct cell-cell communication (investigated by using partial dish irradiations) or by medium borne factors (a co-culture system where two monolayers share the same medium but only one monolayer is exposed to ionising radiation). CDKN1A was found to be up regulated in both directly exposed and non-exposed cells. The data suggest that direct cell-cell communication dominates for these confluent cells, with medium borne factors also contributing. PMID- 17132671 TI - Auger electrons--a nanoprobe for structural, molecular and cellular processes. AB - This paper provides a brief review of recently published work on biophysical and biological aspects of Auger processes. Three specific questions have been considered. (1) Does charge neutralisation contribute to molecular damage such as DNA strand breaks? (2) How many DNA double strand breaks are produced by a single decay of DNA bound (125)I? (3) What is the correlation between number of gammaH2AX foci and number of double strand breaks (DSB)? The paper also gives preliminary reports on two new calculations: (a) calculation of the spectrum of Auger electrons released during decay of (124)I and (b) the use of Auger electrons in the decay of (125)I as a probing agent of novel DNA structures. PMID- 17132672 TI - Heavy ion track structure simulations in liquid water at relativistic energies. AB - Interaction cross sections for bare heavy charged (HZE) particles are obtained from proton interaction cross sections by scaling laws. Proton interaction cross sections are calculated within the (relativistic) plane wave Born approximation and the modelled dielectric response function of liquid water. Relativistic polarisation effects (Fermi density effect) are discussed. The interaction model is implemented into the biophysical track structure simulation code PARTRAC. PMID- 17132673 TI - Single-ion dosemeter based on floating gate memories. AB - Floating Gate (FG) nonvolatile memories are based on a tiny polysilicon layer (the FG) which can be permanently charged with electrons or holes, thus changing the threshold voltage of a MOSFET. Every time a FG is hit by a high energy ion, it experiences a charge loss, depending on the ion linear energy transfer (LET) and on the transistor geometrical and electrical characteristics. This paper discusses the opportunities to use this devices as single an ion dosemeter with sub-micrometer spatial resolution and capable of distinguish the impinging ion LET. PMID- 17132674 TI - Dose-rate dependent effects of ionizing radiation on vascular reactivity. AB - This study was designed to investigate the dose-rate dependent effects of ionising radiation on endothelium- and NO-mediated reactivity of aorta and coronary vessels. Rats were exposed to acute ((137)Cs, 9 x 10(-4) Gy s(-1), 18 min) and chronic ((137)Cs, 2.8 x 10(-7) Gy s(-1), 41 days) radiation in 1 Gy dose. Acute irradiation transiently increased coronary flow in eNOS-activity dependent manner on day 3 after exposure. In striking contrast, chronic irradiation caused a significant depression of coronary flow even on day 90 after irradiation and abolished the effects of NO-synthase inhibitor N-nitro-L-arginine methyl ester (10 micromol l(-1)). Furthermore, low intensity radiation strongly diminished the vasodilator properties of NO-donor sodium nitroprusside (5 micromol l(-1)). A similar pattern was observed in aortic rings. Endothelium dependent vasodilation was increased on days 3 and 10 after acute irradiation, but strongly inhibited following chronic exposure for the entire post-radiation period. This was accompanied by a diminished vasodilator response to NO-donor on days 3, 10 and 30 of post-radiation but not on day 90. The data suggest that ionising radiation in 1 Gy induces changes of aortic and coronary vessels reactivity depending on the dose-rate and the interval after exposure. PMID- 17132675 TI - Evidence of low-dose hyper-radiosensitivity in normal cells of cervix cancer patients? AB - The aim of the present study was to examine the low-dose radiation response of human normal cells using the micronucleus assay. Skin fibroblasts and keratinocytes derived from 40 cervix cancer patients were studied. After in vitro gamma irradiation with single doses ranging from 0.05 to 4 Gy, the fraction of binucleated cells with micronuclei was assessed. For each patient, the Linear Quadratic (LQ) model and the Induced-Repair (IR) model were fitted over the whole data set (0.05-4 Gy). In conclusion, the present study showed some evidence of low-dose hypersensitivity in the fibroblasts of two patients and in the keratinocytes of four of the forty patients studied. PMID- 17132676 TI - Prediction of dose response for radiation induced exchange aberrations taking cell cycle delays into account. AB - Chromosomal aberrations (CAs) are regarded as one of the most sensitive biological indicators of genetic alterations. The aberration frequency is routinely determined in the first metaphase. Yet, the data interpretation can be complicated due to radiation induced mitotic delays. To investigate the effect of delays on CA frequency in the first mitosis, human lymphocytes were irradiated with X rays and Giemsa detectable CAs were measured at different sampling times. Besides, a computer simulation was performed reproducing the main effects under investigation, that is, CA induction and cell progression through the mitotic cycle. The CA formation model takes into account the structural organisation of interphase chromosomes in a lymphocyte nucleus, DNA double-strand break (DSB) induction and their rejoining/misrejoining. Lymphocyte transition through the cell cycle was simulated by a Monte Carlo technique. The delay was proposed to result from DNA DSBs. The predicted ratios of first/second/third cycle metaphases agree with the experimental data for control and irradiated samples. Both experimental and calculated CA frequencies in the first mitosis were nearly time independent. This was proposed to result from de-synchronisation of the lymphocyte population. PMID- 17132677 TI - ESHRE PGD Consortium data collection VI: cycles from January to December 2003 with pregnancy follow-up to October 2004. AB - The sixth report of the ESHRE PGD Consortium is presented, relating to cycles collected for the calendar year 2003 and follow-up of the pregnancies and babies born up to October 2004. Since the beginning of the data collections, there has been a steady rise in the number of cycles, pregnancies and babies reported. For this report, 50 centres participated, reporting on 2984 cycles, 501 pregnancies and 373 babies born. Five hundred and twenty-nine cycles were reported for chromosomal abnormalities, 516 cycles were reported for monogenic diseases, 137 cycles were reported for sexing for X-linked diseases, 1722 cycles were reported for preimplantation genetic screening (PGS) and 80 cycles were reported for social sexing. Data VI is compared to the cumulative data for data collections I V. PMID- 17132678 TI - Effects of fortified milk on morbidity in young children in north India: community based, randomised, double masked placebo controlled trial. AB - OBJECTIVE: To evaluate the efficacy of milk fortified with specific multiple micronutrients on morbidity in children compared with the same milk without fortification. DESIGN: Community based, double masked, individually randomised trial. SETTING: Peri-urban settlement in north India. PARTICIPANTS: Children (n=633) aged 1-3 randomly allocated to receive fortified milk (n=316) or control milk (n=317). INTERVENTION: One year of fortified milk providing additional 7.8 mg zinc, 9.6 mg iron, 4.2 microg selenium, 0.27 mg copper, 156 microg vitamin A, 40.2 mg vitamin C, 7.5 mg vitamin E per day (three feeds). MAIN OUTCOME MEASURES: Days with severe illnesses, incidence and prevalence of diarrhoea, and acute lower respiratory illness. RESULTS: Study groups were comparable at baseline; compliance in the groups was similar. Mean number of episodes of diarrhoea per child was 4.46 (SD 3.8) in the intervention (fortified milk) group and 5.36 (SD 4.1) in the control group. Mean number of episodes of acute lower respiratory illness was 0.62 (SD 1.1) and 0.83 (SD 1.4), respectively. The fortified milk reduced the odds for days with severe illnesses by 15% (95% confidence interval 5% to 24%), the incidence of diarrhoea by 18% (7% to 27%), and the incidence of acute lower respiratory illness by 26% (3% to 43%). Consistently greater beneficial effects were observed in children aged < or =24 months than in older children. CONCLUSION: Milk is well accepted as a means of delivery of micronutrients. Consumption of milk fortified with specific micronutrients can significantly reduce the burden of common morbidities among preschool children, especially in the first two years of life. TRIAL REGISTRATION: NCT00255385 [ClinicalTrials.gov]. PMID- 17132679 TI - Effect of a centralised transfer service on characteristics of inter-hospital neonatal transfers. AB - OBJECTIVE: To determine the effect of a centralised neonatal transfer service on numbers of neonatal transfers and the time taken for teams to reach the baby. DESIGN: Prospective census of neonatal inter-hospital transfers between May and July 2004. Comparison with a previous census undertaken before introduction of the service. Analysis of requests for antenatal in-utero transfer to the regional emergency bed service. SETTING: Geographically defined area in London and southeast England. PATIENTS: Babies transferred to or from a neonatal unit. INTERVENTIONS: Introduction of a centralised neonatal transfer service. MAIN OUTCOME MEASURES: Numbers of transfers, time taken for teams to arrive to the baby (response time). RESULTS: During the census there were 835 transfers with an increase of 34% from the previous census (n = 619). Most of the increase was in urgent transfers for neonatal intensive care. There was a mean of 4.4 urgent transfers a day, with 3.9 elective and 0.8 short-term transfers. Over the same period in-utero transfers decreased. Response times improved from a median of 2 h in 2001 to 1.45 h in 2004 (p<0.05). The 90th centile fell from 6 h to 4.9 h. CONCLUSION: Following the introduction of a centralised neonatal transfer service, response times improved significantly. An increase in the numbers of transfers for medical intensive care was associated with a reduced number of in utero transfers. To balance the improved safety and accessibility of neonatal transfer, similar developments may be needed to facilitate in-utero transfer. PMID- 17132680 TI - Persistent lactic acidosis in neonatal hypoxic-ischaemic encephalopathy correlates with EEG grade and electrographic seizure burden. AB - BACKGROUND: Predicting at birth which infants with perinatal hypoxic-ischaemic injury will progress to significant encephalopathy remains a challenge. OBJECTIVE: To determine whether lactic acidosis at birth in asphyxiated neonates could predict the grade of EEG encephalopathy by examining the relationship between time taken for the normalisation of lactate, severity of encephalopathy and seizure burden. METHODS: Continuous early video-EEG monitoring was performed in babies at risk for hypoxic-ischaemic encephalopathy. Encephalopathy was graded from the EEG data. Total seizure burden (seconds) was calculated for each baby. Initial blood gas measurements of pH, base deficit and lactate were taken within 30 minutes of delivery. Time to normal serum lactate was determined in hours from birth for each infant. RESULTS: All 50 term infants had raised initial serum lactate (median (lower, upper quartiles) 11.7 (10.2, 14.9)). There were no significant differences between the initial serum lactate, pH and base deficit in infants with normal/mildly abnormal (n = 24), moderately abnormal (n = 14), severely abnormal (n = 5) and inactive EEGs (n = 7). Time to normal lactate varied significantly with EEG grade (median (lower, upper quartile) 6.0 (4.1, 9.5) for mild/normal EEG, 13.5 (6.8, 23.5) moderate EEG, 41.5 (30.0, 55.5) severe group, 12.0 (8.1, 21.5) inactive group; p<0.001). Time to normal lactate correlated significantly with EEG seizure burden (seconds; R = 0.446, p = 0.002). Mean (SD) time to normal lactate was 10.0 (7.2) hours in infants who did not have seizures and 27.3 (19.0) hours in the 13 infants with electrographic seizures (p = 0.002). CONCLUSIONS: Serum lactate levels in the first 30 minutes of life do not predict the severity of the ensuing encephalopathy. In contrast, sustained lactic acidosis is associated with severe encephalopathy on EEG and correlates with seizure burden. PMID- 17132681 TI - Mathematical analysis of a free-boundary model for lung branching morphogenesis. AB - Lung branching morphogenesis has been widely studied in the field of developmental biology. Lung airway trees consist of relatively regular-sized distal branches, but how this regular branched pattern is formed is not well understood. In the present study, we undertake a detailed mathematical analysis of the model proposed in (Hartmann & Miura (2006), which numerically captures branching morphogenesis of the simplest possible experimental system in vitro. We investigate analytically the stability of 1D travelling waves with respect to periodic perturbations in two dimensions. This linear stability analysis leads to the so-called dispersion relations, predicting that a certain representative length dominates in this model. As the analytical analysis is restricted to travelling waves, we generalize the linear analysis to any 1D solution by numerical simulations. Both results predict how the representative lengths will change by experimentally changing specific parameters. Finally, we discuss the importance of the analytical results from a biological point of view and propose an experimental scheme for a quantitative comparison between experiments and theory. PMID- 17132682 TI - Spectral analysis of two-signed microarray expression data. AB - We give a simple and informative derivation of a spectral algorithm for clustering and reordering complementary DNA microarray expression data. Here, expression levels of a set of genes are recorded simultaneously across a number of samples, with a positive weight reflecting up-regulation and a negative weight reflecting down-regulation. We give theoretical support for the algorithm based on a biologically justified hypothesis about the structure of the data, and illustrate its use on public domain data in the context of unsupervised tumour classification. The algorithm is derived by considering a discrete optimization problem and then relaxing to the continuous realm. We prove that in the case where the data have an inherent 'checkerboard' sign pattern, the algorithm will automatically reveal that pattern. Further, our derivation shows that the algorithm may be regarded as imposing a random graph model on the expression levels and then clustering from a maximum likelihood perspective. This indicates that the output will be tolerant to perturbations and will reveal 'near checkerboard' patterns when these are present in the data. It is interesting to note that the checkerboard structure is revealed by the first (dominant) singular vectors--previous work on spectral methods has focussed on the case of nonnegative edge weights, where only the second and higher singular vectors are relevant. We illustrate the algorithm on real and synthetic data, and then use it in a tumour classification context on three different cancer data sets. Our results show that respecting the two-signed nature of the data (thereby distinguishing between up-regulation and down-regulation) reveals structures that cannot be gleaned from the absolute value data (where up- and down-regulation are both regarded as 'changes'). PMID- 17132683 TI - Why is the distribution of HTLV-I carriers geographically biased? An answer through a mathematical epidemic model. AB - Human T-cell leukemia virus type I (HTLV-I) is a retrovirus that causes adult T cell leukemia, and the distribution of HTLV-I carriers is endemically biased, e.g. in Japan the density of carriers is high in Kyushu and Okinawa. In order to consider population dynamics over long times taking account of an increase and a decrease of a population we propose a continuous-time HTLV-I model. The model describes population dynamics of carrier numbers and that of carrier proportions, and theoretical results about an increase and a decrease of carriers are obtained. The present approach derives an explanation of the biased distribution of carriers in Japan. PMID- 17132684 TI - Novel G423S mutation of human alpha7 nicotinic receptor promotes agonist-induced desensitization by a protein kinase C-dependent mechanism. AB - The alpha7 nicotinic acetylcholine receptor subunit (CHRNA7) gene harbors a high degree of polymorphism. In this study, we found a novel variant (1267 G to A) in exon 10 of the CHRNA7 gene in a Japanese population. This variant results in glycine-to-serine substitution at position 423 (G423S) located in the large cytoplasmic loop of the protein. To clarify the possibility that the G423S mutation alters the pharmacological properties of alpha7 receptors, acetylcholine (ACh)-elicited current through alpha7-G423S mutant receptors expressed in Xenopus laevis oocytes was measured using the two-electrode voltage-clamp technique. We found that the current elicited by ACh (1 mM, 5 s) through alpha7-G423S receptors, but not through alpha7 receptors, was significantly decreased by treatment with a protein kinase C activator, phorbol-12-myristate-13-acetate (PMA, 10-30 nM). In addition, PMA (10 nM) selectively promoted a progressive decrease in alpha7-G423S current induced by repetitive application of ACh pulses (1 mM, 0.1 s, 0.17-0.33 Hz) compared with alpha7 current. PMA also enhanced the inactivation of alpha7-G423S mutant receptors induced by a prolonged application of choline (30 microM) without affecting alpha7 receptor responses. Western blot analysis showed that the treatment with PMA (30 nM) increased the serine phosphorylation level of the alpha7-G423S mutant receptors but not that of the wild-type receptors. These findings demonstrate that the G423S mutation promotes receptor desensitization by a protein kinase C-dependent mechanism. Thus, we provide the first evidence that a variant in the human CHRNA7 gene alters the function of alpha7 nicotinic receptors. PMID- 17132685 TI - Ca2+ permeability of the (alpha4)3(beta2)2 stoichiometry greatly exceeds that of (alpha4)2(beta2)3 human acetylcholine receptors. AB - Human alpha4beta2 nicotinic acetylcholine receptors (AChRs) expressed in Xenopus laevis oocytes or transfected cell lines are present as a mixture of two stoichiometries, (alpha4)2(beta2)3 and (alpha4)3(beta2)2, which differ depending on whether a beta2 or alpha4 subunit occupies the accessory subunit position corresponding to beta1 subunits of muscle AChRs. Pure populations of each stoichiometry can be expressed in oocytes by combining a linked pair of alpha4 and beta2 with free beta2 to produce the (alpha4)2(beta2)3 stoichiometry or with free alpha4 to produce the (alpha4)3(beta2)2 stoichiometry. We show that the (alpha4)3(beta2)2 stoichiometry and the (alpha4)2(beta2)2beta3 and (alpha4)2(beta2)2alpha5 subtypes in which beta3 or alpha5occupy the accessory positions have much higher permeability to Ca2+ than does (alpha4)2(beta2)3 and suggest that this could be physiologically significant in triggering signaling cascades if this stoichiometry or these subtypes were found in vivo. We show that Ca2+ permeability is determined by charged amino acids at the extracellular end of the M2 transmembrane domain, which could form a ring of amino acids at the outer end of the cation channel. Alpha4, alpha5, and beta3 subunits all have a homologous glutamate in M2 that contributes to high Ca2+ permeability, whereas beta2 has a lysine at this position. Subunit combinations or single amino acids changes at this ring that have all negative charges or a mixture of positive and negative charged amino acids are permeable to Ca2+. All positive charges in the ring prevent Ca2+ permeability. Increasing the proportion of negative charges is associated with increasing permeability to Ca2+. PMID- 17132686 TI - Tumor necrosis factor alpha and endothelin-1 increase P-glycoprotein expression and transport activity at the blood-brain barrier. AB - The ATP-driven drug efflux pump, P-glycoprotein, is a critical and selective element of the blood-brain barrier and a primary impediment to pharmacotherapy of central nervous system (CNS) disorders. Thus, an understanding of how P glycoprotein function is regulated has the potential to improve CNS therapy. We recently demonstrated rapid (minutes) and reversible inactivation of P glycoprotein in rat brain capillaries signaled through tumor necrosis factor alpha (TNF-alpha) and endothelin-1 (ET-1), components of the brain's innate immune response. In this study, we examined the longer-term consequences of continuous exposure of rat brain capillaries to low levels of TNF-alpha and ET-1. Exposing brain capillaries to TNF-alpha or ET-1 caused a rapid decrease in P glycoprotein transport activity with no change in transporter protein expression. This was followed by a 2- to 3-h plateau at the low activity level and then by a sharp increase in both transport activity and protein expression. After 6 h, transport activity and transporter protein expression was double that of control samples. TNF-alpha signaled through TNF-R1, which in turn caused ET release and action through ETA and ETB receptors, nitric-oxide synthase, protein kinase C and nuclear factor-kappaB (NF-kappaB) and finally increased P-glycoprotein expression and transport activity. Assuming similar effects occur in vivo, the present results imply a tightening of the selective blood-brain barrier with chronic inflammation and thus reduced efficacy of CNS-acting drugs that are P glycoprotein substrates. Moreover, involvement of NF-kappaB raises the possibility that other effectors acting through this transcription factor may have similar effects on this key blood-brain barrier transporter. PMID- 17132687 TI - Mutational disruption of a conserved disulfide bond in muscarinic acetylcholine receptors attenuates positive homotropic cooperativity between multiple allosteric sites and has subtype-dependent effects on the affinities of muscarinic allosteric ligands. AB - The 2nd outer loop (o2) of muscarinic acetylcholine receptors (mAChRs) contains a highly conserved cysteine residue that is believed to participate in a disulfide bond and is flanked on either side by epitopes that are critical to the binding of many muscarinic allosteric modulators. We determined the allosteric binding parameters of the modulators gallamine, W84, and tetrahydroaminoacridine (THA) at M2 and M3 mAChRs in which these cysteine residues had been mutated to alanines. THA is known to bind to mAChRs with a strong positive homotropic cooperativity (a Hill slope of approximately 2) that implies that it must interact with multiple allosteric sites. The disulfide cysteine mutations in M2 receptors reduced the allosteric potencies of the tested modulators as if the critical adjacent residue (Tyr177) itself had been mutated. However, in M3 receptors, the disulfide cysteine mutations had no effect on the potencies of gallamine or W84 and even increased the potency of THA. It was most interesting that the strong, positive, homotropic interactions of THA at both M2 and M3 receptors were markedly reduced by the cysteine mutations. In addition, gallamine also displayed positive homotropic cooperativity in its interactions with M3 receptors (but not M2 receptors), and this cooperativity was not evident in the cysteine mutants. Thus, it seems that these cysteine residues play a role in linking cooperating allosteric sites, although it is not currently possible to say whether these multiple sites lie within one receptor or on two linked receptors of a dimer or higher order oligomer. PMID- 17132688 TI - Modulating the folding of P-glycoprotein and cystic fibrosis transmembrane conductance regulator truncation mutants with pharmacological chaperones. AB - Cystic fibrosis transmembrane conductance regulator (CFTR) and P-glycoprotein (P gp) are ATP-binding cassette (ABC) transporters that have two transmembrane domains (TMDs) and two nucleotide-binding domains (NBDs). Defective folding of CFTR lacking phenylalanine 508 (DeltaPhe508) in NBD1 is the most common cause of cystic fibrosis. The Phe508 position seems to be universally important in ABC transporters because deletion of the equivalent residue (Tyr490) in P-gp also inhibits maturation of the protein. The pharmacological chaperone VRT-325 can repair the DeltaPhe508-type folding defects in P-gp or CFTR. VRT-325 may repair the folding defects by promoting dimerization of the two NBDs or by promoting folding of the TMDs. To distinguish between these two mechanisms, we tested the ability of VRT-325 to promote folding of truncation mutants lacking one or both NBDs. Sensitivity to glycosidases was used as an indirect indicator of folding. It was found that VRT-325 could promote maturation of truncation mutants lacking NBD2. Truncation mutants of CFTR or P-gp lacking both NBDs showed deficiencies in core-glycosylation that could be partially reversed by carrying out expression in the presence of VRT-325. The results show that dimerization of the two NBDs to form a "nucleotide-sandwich" structure or NBD interactions with the TMDs are not essential for VRT-325 enhancement of folding. Instead, VRT-325 can promote folding of the TMDs alone. The ability of VRT-325 to promote core-glycosylation of the NBD-less truncation mutants suggests that one mechanism whereby the compound enhances folding is by promoting proper insertion of TM segments attached to the glycosylated loops so that they adopt an orientation favorable for glycosylation. PMID- 17132689 TI - Inhibition of the c-Jun N-terminal kinase-mediated mitochondrial cell death pathway restores auditory function in sound-exposed animals. AB - We tested and characterized the therapeutic value of round window membrane delivered (RWM) d-JNKI-1 peptide (Bonny et al., 2001) against sound trauma induced hearing loss. Morphological characteristics of sound-damaged hair cell nuclei labeled by Hoechst staining show that apoptosis is the predominant mode of cell death after sound trauma. Analysis of the events occurring after sound trauma demonstrates that c-Jun N-terminal kinase (JNK)/stress-activated protein kinase activates a mitochondrial cell death pathway (i.e., activation of Bax, release of cytochrome c, activation of procaspases, and cleavage of fodrin). Fluorescein isothiocyanate (FITC)-conjugated d-JNKI-1 peptide applied onto an intact cochlear RWM diffuses through this membrane and penetrates cochlear tissues with the exception of the stria vascularis. A time sequence of fluorescence measurements demonstrates that FITC-labeled d-JNKI-1 remains in cochlear tissues for as long as 3 weeks. In addition to blocking JNK-mediated activation of a mitochondrial cell death pathway, RWM-delivered d-JNKI-1 prevents hair cell death and development of a permanent shift in hearing threshold that is caused by sound trauma in a dose-dependent manner (EC50 = 2.05 microM). The therapeutic window for protection of the cochlea from sound trauma with RWM delivery of d-JNKI-1 extended out to 12 h after sound exposure. These results show that the mitogen-activated protein kinase/JNK signaling pathway plays a crucial role in sound trauma-initiated hair cell death. Blocking this signaling pathway with RWM delivery of d-JNKI-1 may have significant therapeutic value as a therapeutic intervention to protect the human cochlea from the effects of sound trauma. PMID- 17132690 TI - Celecoxib and CVS risk--lessons from the APC and PreSAP studies. PMID- 17132691 TI - Cavitating pneumonia, a severe complication of leflunomide therapy in chronic polyarthritis. PMID- 17132692 TI - CD95-Mediated control of anti-citrullinated protein/peptides antibodies (ACPA) producing plasma cells occurring in rheumatoid arthritis inflamed joints. AB - OBJECTIVE: Serum anti-citrullinated protein/peptides antibodies (ACPA) are a valuable diagnostic parameter that might be involved in rheumatoid arthritis (RA) pathogenesis. CD95-dependent apoptosis is defective in RA synovium. The present study explores the occurrence of ACPA IgG, and the CD95-mediated control of ACPA IgG-secreting plasma cells (PC) in RA patients. METHODS: Mononuclear cells (MC) were purified from synovial fluid (SF) and peripheral blood (PB) of 15 RA patients. PC capable of secreting ACPA IgG were detected in MC cultures. ACPA IgG present in serum and SF, and PB and SF MC culture supernatant was measured by ELISA. CD95, CD27 and CD138 expression was examined on RA PC identified as CD19(low) CD38(high) cells by flow cytometry. CD95-ligation was obtained by treatment of cultured MC with the anti-CD95 Ab CH11. Apoptotic PC were identified as Annexin-V+. RESULTS: ACPA IgG level was found higher in patients' SF than in their serum. PC were detectable in SF and PB, and exhibited high CD95 and CD27 expression. In contrast, SF, but not PB, PC expressed elevated levels of CD138. SF, but not PB, PC actively secreted ACPA IgG in cultures, in a linear fashion for at least 14 days, and CD95-ligation markedly reduced this activity and provoked PC apoptosis. CONCLUSIONS: The results suggest that RA synovium is a prominent site for ACPA IgG formation and for the accumulation of ACPA IgG secreting PC exhibiting prolonged survival, probably due to RA defective CD95 mediated control. PMID- 17132694 TI - Systemic lupus erythematosus in a multiethnic US cohort (LUMINA) XL II: factors predictive of new or worsening proteinuria. AB - OBJECTIVES: To determine the factors predictive of new or worsening proteinuria in a large multiethnic cohort of patients with systemic lupus erythematosus (SLE). METHODS: Five hundred and twenty-nine SLE patients from a multiethnic US cohort [LUpus in MInorities: NAture versus Nurture (LUMINA)] were evaluated for new or worsening proteinuria using the categories of the Systemic Lupus Activity Measure-Revised: (1), normal; (2), trace or 1+ proteinuria on the dipstick; (3), 2-3+ proteinuria and (4), > or =4+ proteinuria. A rise in urinary protein was considered a positive event visit. Basic demographic and socioeconomic variables were assessed at baseline (T0). Clinical and immunological variables including disease features, activity, duration, comorbidities (such as hypertension and diabetes), medications and autoantibodies were assessed at the visit preceding a positive event visit. Selected HLA-DR and HLA-DQ alleles, and FCGR receptor polymorphisms were assessed. Data were analysed using logistic regression analyses and generalized estimating equations. RESULTS: There were 243 patients (59.1% of 93 Texan Hispanics, 37.0% of 100 Puerto Rican Hispanics, 58.0% of 181 African Americans and 29.7% of 155 Caucasians) with new or worsening proteinuria, and 364 positive events in 2801 visits. Younger age [Odds ratio (OR) = 1.013, 95% confidence limits (CL) = 1.001-1.024, P < 0.0334], anti-dsDNA (OR = 1.554, CL = 1.149-2.100, P < 0.0042), and HLA-DRB1*1503 (OR = 1.746, 95% CL = 1.573-2.2673, P < 0.0103) were found to independently predict the occurrence of new or worsening proteinuria. CONCLUSION: The factors predictive of new or worsening proteinuria include traditional factors associated with lupus nephritis, such as age and anti dsDNA, as well as HLA-DRB1*1503, which has not been previously described in association with lupus nephritis, new or worsening proteinuria. PMID- 17132695 TI - Breast cancer survival and tumor characteristics in premenopausal women carrying the CHEK2*1100delC germline mutation. AB - PURPOSE: Women carrying a CHEK2*1100delC germline mutation have an increased risk of developing breast cancer. This study aims to determine the proportion of CHEK2*1100delC carriers in a premenopausal breast cancer population, unselected for family history of breast cancer, and to investigate tumor characteristics and disease outcome with sufficient follow-up. PATIENTS AND METHODS: We identified a retrospective cohort of 1,479 patients, who received surgery for invasive breast cancer between 1970 and 1994. All patients were diagnosed before age 50. Paraffin embedded tissue blocks were collected for DNA isolation (normal tissue), subsequent CHEK2*1100delC analysis, and tumor revision. Median follow-up was 10.1 years. RESULTS: We detected a CHEK2*1100delC germline mutation in 54 patients (3.7%). Tumor characteristics of CHEK2*1100delC carriers did not differ significantly from those of noncarriers. CHEK2*1100delC carriers had a two-fold increased risk (hazard ratio [HR], 2.1; 95% CI, 1.0 to 4.3; P = .049) of developing a second breast cancer and they had worse recurrence-free survival (HR, 1.7; 95% CI, 1.2 to 2.4; P = .006) and worse breast cancer-specific survival (HR, 1.4; 95% CI, 1.0 to 2.1; P = .072) compared with noncarriers. The poorer disease outcome of CHEK2*1100delC carriers could not be explained by the increased risk of second breast cancer. CONCLUSION: Our study, which is representative for the premenopausal breast cancer population, reveals approximately 4% CHEK2*1100delC carriers have an increased risk of second breast cancer and a worse long-term recurrence-free survival rate. Their identification at time of diagnosis and prolonged intensive follow-up should be considered to optimize clinical management. PMID- 17132696 TI - CHEK2 mutation and hereditary breast cancer. PMID- 17132697 TI - Use of oral conjugated estrogen alone and risk of breast cancer. AB - The authors conducted a prospective cohort analysis in the Women's Health Study (1992-2004), a completed randomized trial assessing aspirin and vitamin E in the primary prevention of cancer and cardiovascular disease, to evaluate use of oral conjugated estrogen alone (0.625 mg/day) and breast cancer risk in a time-varying fashion. Over an average of 10 years of follow-up, 305 incident cases of breast cancer (258 invasive and 47 in situ cancers) were documented among 12,718 women aged 45 years or more who were either consistent current users of oral conjugated estrogen alone (0.625 mg/day) or never users of any type of postmenopausal hormones at baseline and during follow-up. The multivariable hazard ratios comparing "consistent current users" with "never users" were 1.11 (95% confidence interval: 0.79, 1.56) for total breast cancer and 1.13 (95% confidence interval: 0.77, 1.64) for invasive cases. No significant associations were observed for use of less than 8 and 8 years or more. Restricting the analyses to women with prior hysterectomy somewhat strengthened the associations, albeit still not significantly. These data, like those from the Women's Health Initiative, show no significant increase in breast cancer risk with use of oral conjugated estrogen alone (0.625 mg/day), but a small increase or decrease in risk cannot be excluded. PMID- 17132698 TI - Factors associated with prevalent and incident urinary incontinence in a cohort of midlife women: a longitudinal analysis of data: study of women's health across the nation. AB - To compare the characteristics of and baseline factors associated with prevalent and incident urinary incontinence in a diverse cohort of midlife women, the authors analyzed the baseline and first five annual follow-up visits of the Study of Women's Health Across the Nation (SWAN), 1995-2001. From responses to annual questionnaires, the authors defined prevalent incontinence as at least monthly incontinence reported at baseline and incident incontinence as at least monthly incontinence first reported over follow-up. They used multiple logistic regression for their comparison. The mean age of their cohort at baseline was 45.8 (standard deviation: 2.7) years. Prevalent incontinence was 46.7%, and the average incidence was 11.1% per year. Most women reported stress, but a higher proportion developed urge incontinence (15.9% vs. 7.6% at baseline). African Americans (29.5%) and Hispanics (27.5%) had the lowest prevalence of incontinence; African Americans (11.6%) and Caucasians (13.4%) had the highest average annual incidence. Parity, diabetes, fibroids, and poor social support were associated with prevalent incontinence, while high body mass index, high symptom sensitivity, and poor health were associated with incident incontinence. In midlife women, incident incontinence is mild with different characteristics and baseline risk factors; overweight women have a higher risk of developing incontinence. PMID- 17132699 TI - Re: risk of second primary malignancies in women with papillary thyroid cancer. PMID- 17132701 TI - Tetramethylpyrazine protects rat renal tubular cell apoptosis induced by gentamicin. AB - BACKGROUND: Gentamicin, a widely used antibiotic for the treatment of bacterial infection, can cause nephrotoxicity. Tetramethylpyrazine (TMP) is a compound purified from the rhizome of Ligusticum wallichi (Chuanxiong) and has been found to protect against ischaemia-reperfusion injury, nephritis and alcohol-induced toxicity in rat kidneys. METHODS: We used rat renal tubular cells (RTCs), NRK 52E, in this study. The cytotoxicity of gentamicin was checked with transferase mediated deoxyuridine triphosphate nick end-labeling (TUNEL) staining, and the generation of reactive oxygen species was measured using the fluorescent probe 2,7-dichlorofluorescein. We evaluated several apoptotic parameters: cleaved caspase levels, tumour necrosis factor (TNF-alpha) excretion and nuclear factor Kappa B (NF-kappaB) activity. We also examined the TMP protective effect on gentamicin-induced apoptosis in rat kidneys. RESULTS: The results of this study showed that gentamicin was found to markedly induce apoptosis in NRK-52E cells in a dose-dependent manner; that TMP expressed a dose-dependent protective effect against gentamicin-induced apoptosis; that pre-treatment of the cells with 50 or 100 microM of TMP effectively decreased the reactive oxygen species formation induced by gentamicin; that TMP was found to inactivate the gentamicin-stimulated activities of caspase-3, caspase-8 and caspase-9, to inhibit gentamicin-induced release of cytochrome c, as well as to raise the expression of Bcl-x(L); that TMP inhibited the gentamicin-induced TNF-alpha excretion, and inactivated the transcription factor NF-kappaB; and that the TMP treatment significantly reduced apoptotic injury in rat RTCs. CONCLUSIONS: Based on the results of this study, we suggest that TMP can attenuate gentamicin-induced oxidative stress and apoptotic injury in rat RTCs, and that its character may have therapeutic potential for patients with renal diseases. PMID- 17132702 TI - The effect of pH-neutral peritoneal dialysis fluids on adipokine secretion from cultured adipocytes. AB - BACKGROUND: Adipokines are a group of fat-secreted hormones and cytokines, including leptin and adiponectin, with important functions in humans. Peritoneal dialysis (PD) is associated with markedly raised plasma adipokines, suggesting increased production in this setting. We have shown that low pH down-regulates leptin production. The current study was designed to test if novel pH-neutral PD fluids may regulate leptin and adiponectin secretion in vitro. METHODS: We exposed 3T3-L1 adipocytes to a 50 : 50 mixture of dialysate and M199 containing 10% serum for upto 48 h. Dialysates were commercial PD fluids, i.e. conventional acidic, lactate-buffered solutions (PD-acid) and pH-neutral lactate-buffered (PD Bal) or bicarbonate-buffered solutions (PD-Bic). Leptin and adiponectin concentrations in culture-cell media were measured by ELISA. RESULTS: Compared with PD-acid, PD-Bal and PD-Bic produced a 25 and 43% increase, respectively, in leptin secretion at 48 h (P < 0.05). In contrast, adiponectin secretion was not affected. High glucose PD fluids (4.25%) specifically inhibited leptin secretion vs 1.5% glucose, buffer-matched solutions (P < 0.05). However, differences in leptin secretion due to pH and type of buffer remained significant. In further experiments, the pH of test media were extensively varied without the presence of dialysates. Leptin secretion was shown to increase in a parallel to pH, whereas large changes in pH did not affect adiponectin secretion. CONCLUSION: The pH neutral PD solutions specifically induce leptin, but not adiponectin secretion from 3T3-L1 adipocytes. PD-Bic produced a greater leptin stimulation than PD-Bal, but this difference was attributable to pH per se, rather than the type of buffer. PMID- 17132703 TI - Human urotensin II in the plasma of anephric subjects. PMID- 17132704 TI - Successful living donor kidney transplantation across HLA and ABO incompatibilities. PMID- 17132705 TI - Impact of calcium and vitamin D therapy on arterial and cardiac disease in young adults with childhood-onset end stage renal disease. PMID- 17132706 TI - New horizons in prevention and treatment of ischaemic injury to kidney transplants. PMID- 17132707 TI - Influence of haemodialysis and left ventricular failure on peripheral A(2A) adenosine receptor expression. AB - BACKGROUND: Haemodialysis (HD) sometimes accelerates left ventricular failure (LVF). As adenosine (ADO) is strongly implicated in cardiovascular functions, particularly via A(2A) receptor activation and as changes of peripheral A(2A) receptors mirror changes occurring in the cardiovascular system, we examined the influence of HD and LVF on both ADO plasma concentration and the expression of A(2A) receptors (i.e. Bmax, K(D) and mRNA amount) of peripheral blood mononuclear cells. METHODS: This cross-sectional study included 61 chronic renal failure (CRF) patients: 41 without LVF (24 haemodialysed and 17 undialysed) and 20 with LVF (9 haemodialysed and 11 undialysed). Ten LVF patients without CRF and 10 healthy subjects were also examined. RESULTS: (i) Bmax values of CRF patients without LVF were significantly decreased in undialysed patients compared with haemodialysed patients, and compared with controls (69 +/- 25 vs 98 +/- 33 vs 180 +/- 60 fmol/mg of protein, P < 0.05). Bmax values of CRF patients with LVF were lower in undialysed patients than in haemodialysed patients (60 +/- 27 vs 101 +/- 27 fmol/mg of protein, P < 0.05). Bmax values of LVF patients without CRF were lower than in controls (51 +/- 19 vs 180 +/- 60 fmol/mg of protein). (ii) A(2A) mRNA expression was increased in haemodialysed patients compared with controls (20.2 +/- 0.75 vs 17.6 +/- 1.3, P < 0.05). (iii) ADO plasma levels were high in haemodialysed patients and further increased during the HD sessions. CONCLUSION: The number of A(2A) receptors was decreased by CRF with or without LVF. However, this decrease was less important in haemodialysed patients. The changes in peripheral A(2A) receptor expression suggest a significant inflammatory response to HD and heart or kidney failure. Whether these changes do reflect alterations in cardiomyocytes needs further investigation. PMID- 17132708 TI - Multiple infections after commercial renal transplantation in India. PMID- 17132709 TI - Isolated hydatid disease of the native kidney in a renal transplant recipient. PMID- 17132710 TI - Effect of L-carnitine administration on erythropoietin use in thalassemic minor haemodialysis patients. PMID- 17132711 TI - Insights on the development, kinetics, and variation of photoinhibition using chlorophyll fluorescence imaging of a chilled, variegated leaf. AB - The effect of chilling on photosystem II (PSII) efficiency was studied in the variegated leaves of Calathea makoyana, in order to gain insight into the causes of chilling-induced photoinhibition. Additionally, a relationship was revealed between (chilling) stress and variation in photosynthesis. Chilling treatments (5 degrees C and 10 degrees C) were performed for different durations (1-7 d) under a moderate irradiance (120 micromol m-2 s-1). The individual leaves were divided into a shaded zone and two illuminated, chilled zones. The leaf tip and sometimes the leaf base were not chilled. Measurements of the dark-adapted Fv/Fm were made on the different leaf zones at the end of the chilling treatment, and then for several days thereafter to monitor recovery. Chilling up to 7 d in the dark did not affect PSII efficiency and visual appearance, whereas chilling in the light caused severe photoinhibition, sometimes followed by leaf necrosis. Photoinhibition increased with the duration of the chilling period, whereas, remarkably, chilling temperature had no effect. In the unchilled leaf tip, photoinhibition also occurred, whereas in the unchilled leaf base it did not. Whatever the leaf zone, photoinhibition became permanent if the mean value dropped below 0.4, although chlorosis and necrosis were associated solely with chilled illuminated tissue. Starch accumulated in the unchilled leaf tip, in contrast to the adjacent chilled irradiated zone. This suggests that photoinhibition was due to a secondary effect in the unchilled leaf tip (sink limitation), whereas it was a direct effect of chilling and irradiance in the chilled illuminated zones. The PSII efficiency and its coefficient of variation showed a unique negative linearity across all leaf zones and different tissue types. The slope of this curve was steeper for chilled leaves than it was for healthy, non-stressed leaves, suggesting that the coefficient of variation may be an important tool for assessing stress in leaves. PMID- 17132712 TI - Identification of stress-responsive genes in an indica rice (Oryza sativa L.) using ESTs generated from drought-stressed seedlings. AB - The impacts of drought on plant growth and development limit cereal crop production worldwide. Rice (Oryza sativa) productivity and production is severely affected due to recurrent droughts in almost all agroecological zones. With the advent of molecular and genomic technologies, emphasis is now placed on understanding the mechanisms of genetic control of the drought-stress response. In order to identify genes associated with water-stress response in rice, ESTs generated from a normalized cDNA library, constructed from drought-stressed leaf tissue of an indica cultivar, Nagina 22 were used. Analysis of 7794 cDNA sequences led to the identification of 5815 rice ESTs. Of these, 334 exhibited no significant sequence homology with any rice ESTs or full-length cDNAs in public databases, indicating that these transcripts are enriched during drought stress. Analysis of these 5815 ESTs led to the identification of 1677 unique sequences. To characterize this drought transcriptome further and to identify candidate genes associated with the drought-stress response, the rice data were compared with those for abiotic stress-induced sequences obtained from expression profiling studies in Arabidopsis, barley, maize, and rice. This comparative analysis identified 589 putative stress-responsive genes (SRGs) that are shared by these diverse plant species. Further, the identified leaf SRGs were compared to expression profiles for a drought-stressed rice panicle library to identify common sequences. Significantly, 125 genes were found to be expressed under drought stress in both tissues. The functional classification of these 125 genes showed that a majority of them are associated with cellular metabolism, signal transduction, and transcriptional regulation. PMID- 17132713 TI - Subchronic urinary bladder effects of muraglitazar in male rats. AB - Muraglitazar, a PPARalpha/gamma dual agonist, was dosed orally to rats once daily for 13 weeks to evaluate urinary and urothelial changes of potential relevance to urinary bladder tumorigenesis. Groups of 17 young or aged rats per sex were fed a normal or 1% NH4Cl-supplemented diet and were dosed with 0, 1, or 50 mg/kg muraglitazar. Lithogenic ions and sediment were profiled from freshly voided urine samples collected 24 h after dosing, and drug exposures were measured. Urinary citrate, oxalate, and epidermal growth factor (EGF) were assayed from 18 h urine collections. Urothelium was assessed by light microscopy, scanning electron microscopy, and BrdU and TUNEL immunohistochemistry. When fed a normal diet, urine pH was higher in males (above 6.5). Urine volume/body weight was greater in females. Urine soluble/total calcium and magnesium and phosphorus/creatinine ratios were lower in male rats fed a normal diet. Urine citrate levels were decreased and oxalate was increased in young male rats treated with 50 mg/kg muraglitazar compared to age/sex/diet-matched controls. No changes in urine sediment were detected 24 h after dosing. In young male rats treated with 50 mg/kg on normal diet, multifocal urothelial necrosis and proliferation were observed, whereas urothelial apoptosis and urine EGF levels were unchanged compared to age/sex/diet-matched controls. Urothelial necrosis and proliferation were not correlated to systemic or urinary drug exposures and were prevented by dietary acidification. These data suggest that muraglitazar associated changes in urine composition predispose to urothelial cytotoxicity and proliferation in the urinary bladder of young male rats and that urine sediment must be profiled at multiple daily timepoints to fully qualify drug-induced changes in urine composition. PMID- 17132714 TI - Gestational PFOA exposure of mice is associated with altered mammary gland development in dams and female offspring. AB - Perfluorooctanoic acid (PFOA), with diverse and widespread commercial and industrial applications, has been detected in human and wildlife sera. Previous mouse studies linked prenatal PFOA exposure to decreased neonatal body weights (BWs) and survival in a dose-dependent manner. To determine whether effects were linked to gestational time of exposure or to subsequent lactational changes, timed-pregnant CD-1 mice were orally dosed with 5 mg PFOA/kg on gestation days (GD) 1-17, 8-17, 12-17, or vehicle on GD 1-17. PFOA exposure had no effect on maternal weight gain or number of live pups born. Mean pup BWs on postnatal day (PND) 1 in all PFOA-exposed groups were significantly reduced and decrements persisted until weaning. Mammary glands from lactating dams and female pups on PND 10 and 20 were scored based on differentiation or developmental stages. A significant reduction in mammary differentiation among dams exposed GD 1-17 or 8 17 was evident on PND 10. On PND 20, delays in normal epithelial involution and alterations in milk protein gene expression were observed. All exposed female pups displayed stunted mammary epithelial branching and growth at PND 10 and 20. While control litters at PND 10 and 20 had average scores of 3.1 and 3.3, respectively, all treated litters had scores of 1.7 or less, with no progression of duct epithelial growth evident over time. BW was an insignificant covariate for these effects. These findings suggest that in addition to gestational exposure, abnormal lactational development of dams may play a role in early growth retardation of developmentally exposed offspring. PMID- 17132715 TI - Comparison of the effects of antibody-coated liposomes, IVIG, and anti-RBC immunotherapy in a murine model of passive chronic immune thrombocytopenia. AB - The present work evaluated antibody-coated liposomes as a new treatment strategy for immune thrombocytopenic purpura (ITP) through the use of a mouse model of the disease. Effects of antimethotrexate antibody (AMI)-coated liposomes and intravenous immunoglobulin (IVIG)-coated liposomes (15, 30, 60 micromol lipid/kg) were compared with the effects of IVIG (0.4, 1, 2 g/kg) and anti-red blood cell (anti-RBC) monoclonal antibody immunotherapy (TER119, 5, 15, 25, and 50 microg/mouse) on MWReg30-induced thrombocytopenia. Each treatment was found to attenuate thrombocytopenia in a dose-dependent manner and, consistent with previous work, IVIG was found to increase antiplatelet antibody clearance in a dose-dependent manner. TER119 demonstrated greater effects on thrombocytopenia relative to other therapies (peak platelet counts: 224% +/- 34% of initial platelet counts for 50 microg TER119/mouse versus 160% +/- 34% for 2 g/kg IVIG, 88% +/- 36% for 60 micromol lipid/kg AMI-coated liposomes, and 80% +/- 25% for 60 micromol lipid/kg IVIG-coated liposomes). However, the effects of TER119 were associated with severe hemolysis, as TER119 decreased RBC counts by approximately 50%. The present work demonstrated that antibody-coated liposomes attenuated thrombocytopenia in this model at a much lower immunoglobulin dose than that required for IVIG effects and, in contrast with TER119, antibody-coated liposomes increased platelet counts without altering RBC counts. PMID- 17132716 TI - Spi-1/PU.1 participates in erythroleukemogenesis by inhibiting apoptosis in cooperation with Epo signaling and by blocking erythroid differentiation. AB - Overexpression of the transcription factor Spi-1/PU.1 in mice leads to acute erythroleukemia characterized by a differentiation block at the proerythroblastic stage. In this study, we made use of a new cellular system allowing us to reach graded expression of Spi-1 in preleukemic cells to dissect mechanisms of Spi-1/ PU-1 in erythroleukemogenesis. This system is based on conditional production of 1 or 2 spi-1-interfering RNAs stably inserted into spi-1 transgenic proerythroblasts. We show that Spi-1 knock-down was sufficient to reinstate the erythroid differentiation program. This differentiation process was associated with an exit from the cell cycle. Evidence is provided that in the presence of erythropoietin (Epo), Spi-1 displays an antiapoptotic role that is independent of its function in blocking erythroid differentiation. Apoptosis inhibited by Spi-1 did not involve activation of the Fas/FasL signaling pathway nor a failure to activate Epo receptor (EpoR). Furthermore, we found that reducing the Spi-1 level yields to ERK dephosphorylation and increased phosphorylation of AKT and STAT5, suggesting that Spi-1 may affect major signaling pathways downstream of the EpoR in erythroid cells. These findings reveal 2 distinct roles for Spi-1 during erythroleukemogenesis: Spi-1 blocks the erythroid differentiation program and acts to impair apoptotic death in cooperation with an Epo signaling. PMID- 17132717 TI - Induction of Th2 type immunity in a mouse system reveals a novel immunoregulatory role of basophils. AB - While production of cytokines such as IL-12 by activated dendritic cells supports development of Th1 type immunity, a source of early IL-4 that is responsible for Th2 immunity is not well understood. We now show that coculture of basophils could promote a robust Th2 differentiation upon stimulation of naive CD4 T cells primarily via IL-4. Th2 promotion by basophils was also observed even when naive CD4 T cells were stimulated in a Th1-promoting condition or when fully differentiated Th1 phenotype effector CD4 T cells were restimulated. IL-4 deficient basophils failed to induce Th2 differentiation but suppressed Th1 differentiation. It was subsequently revealed that the IL-4-deficient basophils must engage cell-to-cell contact to exert the inhibitory effect on Th1 differentiation. Stimulation of naive CD4 T cells within an in vivo environment of increased basophil generation supported development of Th2 type immunity. Taken together, our results suggest that basophils may provide an important link for the development of Th2 immunity. PMID- 17132718 TI - A novel human B cell subpopulation representing the initial germinal center population to express AID. AB - We have identified a novel mature human B-cell subpopulation in the human tonsil that has characteristics of both naive B cells and germinal center B cells including the expression of activation-induced cytidine deaminase (AID), which is essential for the process of immunoglobulin somatic hypermutation and class switch recombination. These cells are clearly somatically hypermutated, albeit modestly. Their phenotype (IgD(+)CD38(-)CD23(-)FSC(hi)CD71(+)) is unique and suggests they may be intermediate between both naive and germinal center cells. Morphologically they are also distinct from other B-cell subpopulations. The evidence presented suggests these cells may be the founder cells of the germinal center reaction (a pro-GC cell) and may be the normal counterpart of the mantle cell lymphoma cell. PMID- 17132719 TI - Results of the randomized international FAB/LMB96 trial for intermediate risk B cell non-Hodgkin lymphoma in children and adolescents: it is possible to reduce treatment for the early responding patients. AB - A previous study (LMB89) of the French Society of Pediatric Oncology for childhood mature B-cell lymphoma (B-NHL) demonstrated a 92% 3-year event-free survival (EFS) for intermediate-risk group B defined as "non-resected" stage II/I and CNS-negative advanced-stage IIV/IV (70% of cases). We performed the FAB/LMB96 trial to assess the possibility of reducing treatment in children/adolescents with intermediate-risk B-NHL without jeopardizing survival. "Early responding" patients (tumor response > 20% at day 7) were randomized in a factorial design between 4 arms, 2 receiving half-dose of cyclophosphamide in the second induction course with cyclophosphamide, Oncovin (vincristine), prednisone, Adriamycin (doxorubicin), methotrexate (COPADM) and 2 not receiving the maintenance course M1. A total of 657 patients were randomized (May 1996 to June 2001) and 637 were analyzed. The analysis showed no significant effect of any of the treatment reductions on EFS and survival. The 4-year EFS was 93.4% and 90.9% in the groups with full-dose and half-dose of cyclophosphamide (RR = 1.3, P = .40) and 91.9% and 92.5% in the groups with and without M1 (RR = 1.01, P = .98). There was no interaction between the 2 treatment reductions or between each treatment reduction and LDH level or histologic subtypes (Burkitt/Burkitt-like or large B cell). Children/adolescents with intermediate-risk B-NHL who have an early response and achieve a complete remission after the first consolidation course can be cured with a 4-course treatment with a total dose of only 3.3 g/m2 cyclophosphamide and 120 mg/m2 doxorubicin. PMID- 17132720 TI - Intensified 12-week CHOP (I-CHOP) plus G-CSF compared with standard 24-week CHOP (CHOP-21) for patients with intermediate-risk aggressive non-Hodgkin lymphoma: a phase 3 trial of the Dutch-Belgian Hemato-Oncology Cooperative Group (HOVON). AB - Optimal dose and timing of CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) chemotherapy for aggressive non-Hodgkin lymphoma (NHL) is still an unresolved issue. We assessed whether dose intensifications with cyclophosphamide and doxorubicin might improve outcome in younger patients with intermediate-risk aggressive NHL. Previously untreated patients were assigned to receive either 8 courses of standard CHOP (n = 239) or 6 courses of intensified (I)-CHOP (n = 238). Although there was a tendency in favor of I-CHOP for overall survival (OS), disease-free survival (DFS), and event-free survival (EFS), the differences were not significant. However, although these analyses were not planned, when the intermediate-risk group was divided into low-intermediate- and high-intermediate risk patients according to the International Prognostic Index (IPI), low intermediate-risk patients had improved 6-year OS (67% vs 52%; P = .05), DFS (58% vs 45%; P = .06), and EFS (41% vs 30%; P = .21) when they were treated with I CHOP compared with standard CHOP. On the other hand, high-intermediate-risk patients seem to have no benefit from I-CHOP. Although clinically relevant side effects occurred more often in the I-CHOP arm, treatment-related mortality was similar. These data suggest that I-CHOP might be preferable to standard CHOP in younger patients with low-intermediate-risk aggressive NHL. PMID- 17132721 TI - Pharmacodynamics and safety of intravenous pegaspargase during remission induction in adults aged 55 years or younger with newly diagnosed acute lymphoblastic leukemia. AB - In contrast to that in children, pharmacokinetic, pharmacodynamic, and safety information on pegaspargase in adults is very limited. We administered a single intravenous dose of pegaspargase (2000 IU/m2) as part of a standard frontline induction regimen to 25 adults with newly diagnosed acute lymphoblastic leukemia (ALL), and obtained serum samples on several time points. The population mean peak serum concentration of asparaginase enzymatic activity was 1 IU/mL, the elimination half-life was 7 days, and the volume of distribution was 2.43 L/m2. After the single dose, asparagine deamination was complete in all patients after 2 hours, and in 100%, 81%, and 44% on days 14, 21, and 28, respectively. A pharmocodynamic correlation model showed minimal enzymatic activity of 0.2 IU/mL for optimal asparagine depletion. The kinetic posthoc analyses demonstrated enzymatic activity for 3 weeks or more. One patient developed neutralizing antiasparaginase antibodies on day 22 after administration. Pegaspargase was well tolerated, with few grade 3/4 side effects. No allergic reactions or pancreatitis were observed. In adults aged 55 years or younger, pegaspargase produces a long duration of asparagine depletion and can be given intravenously, with a safety profile that is similar to equivalent multiple doses of intramuscular Escherichia coli asparaginase. PMID- 17132722 TI - Immunostimulatory RNA oligonucleotides trigger an antigen-specific cytotoxic T cell and IgG2a response. AB - Single-stranded RNA oligonucleotides containing an immunostimulatory motif (immunostimulatory RNA [isRNA]) are potent inducers of interferon-alpha via the Toll-like receptor 7. We investigated the effect of isRNA on the development of an immune response. We show that isRNA activates dendritic cells and induces production of Th1-type cytokines both in vitro and in vivo. Cytokine production led to bystander activation of T and B cells. We further demonstrate that isRNA triggers the generation of antigen-specific cytotoxic T cells and of an IgG2a biased antibody response to antigen in a sequence-dependent manner. In summary, we provide evidence for the first time that isRNA oligonucleotides can simultaneously activate the innate and adaptive arms of the immune system. PMID- 17132723 TI - HIV-1 infection and pathogenesis in a novel humanized mouse model. AB - The Rag2-gammaC double-knockout (DKO) mouse lacks T, B, and natural killer (NK) cells, and allows development of a functional human immune system with human CD34+ hematopoietic stem/progenitor cells (DKO-hu HSCs). Normal human T, B, and dendritic cells are present in peripheral blood, thymus, spleen, and lymph nodes. We report that both CCR5 and CXCR4 are expressed on human immature and mature T cells. DKO-hu HSC mice allow efficient HIV-1 infection with plasma high viremia. High levels of productive infection occur in the thymus, spleen, and lymph nodes. Human CD4+ T cells are gradually depleted by HIV-1 in a dose-dependent manner. In addition, HIV-1 infection persists in infected DKO-hu HSC mice for at least 19 weeks, with infectious HIV-1 in lymphoid tissues. Thus, the DKO-hu HSC mouse can serve as a relevant in vivo model to investigate mechanisms of HIV-1 infection and immunopathogenesis as well as to develop anti-HIV-1 therapeutics. PMID- 17132724 TI - Donor cell leukemia: insight into cancer stem cells and the stem cell niche. AB - Donor cell leukemia (DCL) is a rare complication of hematopoietic cell transplantation (HCT). Its incidence has been reported between 0.12% and 5%, although the majority of cases are anecdotal. The mechanisms of leukemogenesis in DCL may be distinct from other types of leukemia. Possible causes of DCL include oncogenic alteration or premature aging of transplanted donor cells in an immunosuppressed person. Although many studies have recently better characterized leukemic stem cells, it is important to also consider that both intrinsic cell factors and external signals from the hematopoietic microenvironment govern the developmental fate of hematopoietic stem cells (HSCs). Therefore, in cases of DCL, alteration of the microenvironment after HCT may increase the likelihood that some progeny of normal HSCs become leukemic. This complex intercommunication between cells, growth factors, and cytokines in the hematopoietic microenvironment are critical to balance HSC self-renewal, proliferation, and differentiation. However, this homeostasis is likely perturbed in the development of DCL, allowing unique insight into the stimuli that regulate normal and potentially abnormal hematopoietic development. In this article, we discuss the possible pathogenesis of DCL, its association with stem cells, and its likely dependence on a less-supportive stem cell niche. PMID- 17132725 TI - Involvement of the fractalkine pathway in the pathogenesis of childhood hemolytic uremic syndrome. AB - Thrombotic microangiopathy and acute renal failure are cardinal features of postdiarrheal hemolytic uremic syndrome (HUS). These conditions are related to endothelial and epithelial cell damage induced by Shiga toxin (Stx) through the interaction with its globotriaosyl ceramide receptor. However, inflammatory processes contribute to the pathogenesis of HUS by sensitizing cells to Stx fractalkine (FKN), a CX(3)C transmembrane chemokine expressed on epithelial and endothelial cells upon activation, is involved in the selective migration and adhesion of specific leukocyte subsets to tissues. Here, we demonstrated a selective depletion of circulating mononuclear leukocytes expressing the receptor for FKN (CX(3)CR1) in patients with HUS. We found a unique phenotype in children with HUS distinct from that seen in healthy, uremic, or infected controls, in which monocytes lost CX(3)CR1, down-modulated CD62L, and increased CD16. In addition, the CD56(dim) natural killer (NK) subpopulation was decreased, leading to an altered peripheral CD56(dim)/CD56(bright) ratio from 10.0 to 4.5. It is noteworthy that a negative correlation existed between the percentage of circulating CX(3)CR1(+) leukocytes and the severity of renal failure. Finally, CX(3)CR1(+) leukocytes were observed in renal biopsies from patients with HUS. We suggest that the interaction of CX(3)CR1(+) cells with FKN present on activated endothelial cells may contribute to renal injury in HUS. PMID- 17132726 TI - Staphylococcal superantigen-like 5 binds PSGL-1 and inhibits P-selectin-mediated neutrophil rolling. AB - Staphylococcus aureus secretes several virulence factors interfering with host cell functions. Staphylococcal superantigen-like (SSL) proteins are a family of 11 exotoxins with structural homology to superantigens but with generally unknown functions. Recently, we described that chemotaxis inhibitory protein of Staphylococcus aureus (CHIPS(31-121)), a potent inhibitor of C5a-induced responses, is structurally homologous to the C-terminal domain of SSL5. Here, we identify P-selectin glycoprotein ligand-1 (PSGL-1), involved in the initial rolling of neutrophils along the endothelium, as a target for SSL5. SSL5 specifically bound to Chinese hamster ovary cells stably expressing PSGL-1 (CHO PSGL-1), which was dependent of sulfation and sialylation. Furthermore, SSL5 bound to PSGL-1/Ig fusion protein immobilized on a biosensor chip. SSL5 affected binding of soluble P-selectin/Fc chimera, the principle ligand of PSGL-1, to CHO PSGL-1 cells and inhibited adhesion of neutrophils to immobilized P-selectin under static conditions. Under flow conditions SSL5 strongly decreased neutrophil rolling on immobilized P-selectin/Fc and activated human endothelial cells. In conclusion, SSL5 interferes with the interaction between PSGL-1 and P-selectin, suggesting that S aureus uses SSL5 to prevent neutrophil extravasation toward the site of infection. This makes SSL5 a potential lead for the development of new anti-inflammatory compounds for disorders characterized by excessive recruitment of leukocytes. PMID- 17132727 TI - Alpha-defensins block the early steps of HIV-1 infection: interference with the binding of gp120 to CD4. AB - Alpha-defensins are antibiotic peptides that act as natural inhibitors of HIV-1 infection. However, the mechanisms of such inhibition are still unclear. Here we demonstrate that alpha-defensins block the earliest steps in the viral infectious cycle, as documented using an HIV-1 envelope-mediated cell-fusion assay. A broad spectrum inhibitory activity was observed on primary and laboratory-adapted HIV-1 isolates irrespective of their coreceptor specificity and genetic subtype. A primary mechanism of such inhibition was identified as the ability of alpha defensins to bind specifically both to the primary HIV-1 cellular receptor, CD4, and to the viral envelope glycoprotein, gp120. Moreover, treatment of CD4+ T cells with alpha-defensins caused a dramatic downmodulation of CD4 expression. By monoclonal antibody competition, the regions of interaction with alpha-defensins were mapped to the D1 domain of CD4 and to a surface contiguous to the CD4- and coreceptor-binding sites of gp120. Consistent with these findings, alpha defensins inhibited the binding of gp120 to CD4. These data demonstrate that alpha-defensins specifically block the initial phase of the HIV infectious cycle and modulate the expression of CD4, a critical receptor in the physiology of T cell activation. PMID- 17132728 TI - Entamoeba histolytica cell movement: a central role for self-generated chemokines and chemorepellents. AB - Entamoeba histolytica cells, the cause of amoebic dysentery, are highly motile, and this motility is an essential feature of the pathogenesis and morbidity of amoebiasis. However, the control of E. histolytica motility within the gut and during invasion is poorly understood. We have used an improved chemotaxis assay to identify the key extracellular signals mediating Entamoeba chemotaxis. The dominant responses we observe are caused by factors generated by E. histolytica cells themselves. Medium that has been conditioned by E. histolytica growth causes both chemokinesis and negative chemotaxis. The speed of random movement is more than doubled in conditioned compared with fresh medium, and cells move efficiently away from conditioned medium by negative chemotaxis. Ethanol, the product of Entamoeba glucose metabolism, is the principal component of the chemokinetic response. The closely related but nonpathogenic Entamoeba dispar shows no change in motility in response to conditioned medium implying that these responses are central to E. histolytica pathogenesis. PMID- 17132729 TI - Smad4 cooperates with lymphoid enhancer-binding factor 1/T cell-specific factor to increase c-myc expression in the absence of TGF-beta signaling. AB - The c-myc protooncogene is a key regulator of cell proliferation whose expression is reduced in normal epithelial cells in response to the growth inhibitory cytokine TGF-beta. Smad4 mediates this inhibitory effect of TGF-beta by forming a complex with Smad3, E2F4/5, and p107 at the TGF-beta inhibitory element (TIE) element on the c-myc promoter. In contrast, cell proliferation and c-myc expression are increased in response to Wnt ligands; this effect is mediated through the lymphoid enhancer-binding factor 1/T cell-specific factor (LEF/TCF) family of transcription factors on the c-myc promoter LEF/TCF-binding elements (TBE1 and TBE2). We report that a peptide aptamer designed to inhibit the binding between Smad4 and LEF/TCF reduced c-myc expression and the growth rate of HepG2 cells. Further analysis demonstrated that, in the absence of TGF-beta, Smad4 was bound to the positive regulatory element TBE1 from the c-myc promoter and activated c-myc promoter activity. Smad4 binding to the positive TBE1 c-myc element was reduced by TGF-beta, consistent with Smad4's inhibitory role on c-myc expression in response to TGF-beta. Reduction of Smad4 levels by RNAi knockdown also reduced c-myc expression levels and sensitized hepatocytes to cell death by serum deprivation. Two tumor-derived mutant Smad4 proteins that fail to mediate TGF-beta responses were still competent to cooperate with LEF1 to activate the c myc promoter. These results support a previously unreported TGF-beta-independent function for Smad4 in cooperating with LEF/TCF to activate c-myc expression. PMID- 17132730 TI - The mediator complex functions as a coactivator for GATA-1 in erythropoiesis via subunit Med1/TRAP220. AB - The Mediator complex forms the bridge between transcriptional activators and RNA polymerase II. Mediator subunit Med1/TRAP220 is a key component of Mediator originally found to associate with nuclear hormone receptors. Med1 deficiency causes lethality at embryonic day 11.5 because of defects in heart and placenta development. Here we show that Med1-deficient 10.5 days postcoitum embryos are anemic but have normal numbers of hematopoietic progenitor cells. Med1-deficient progenitor cells have a defect in forming erythroid burst-forming units (BFU-E) and colony-forming units (CFU-E), but not in forming myeloid colonies. At the molecular level, we demonstrate that Med1 interacts physically with the erythroid master regulator GATA-1. In transcription assays, Med1 deficiency leads to a defect in GATA-1-mediated transactivation. In chromatin immunoprecipitation experiments, we find Mediator components at GATA-1-occupied enhancer sites. Thus, we conclude that Mediator subunit Med1 acts as a pivotal coactivator for GATA-1 in erythroid development. PMID- 17132731 TI - Zinc binding to the HCCH motif of HIV-1 virion infectivity factor induces a conformational change that mediates protein-protein interactions. AB - Virion infectivity factor (Vif) is an accessory protein encoded by HIV-1 and is critical for viral infection of the host CD4(+) T cell population. Vif induces ubiquitination and subsequent degradation of Apo3G, a cytosolic cytidine deaminase that otherwise targets the retroviral genome. Interaction of Vif with the cellular Cullin5-based E3 ubiquitin ligase requires a conserved BC box and upstream residues that are part of the conserved H-(Xaa)(5)-C-(Xaa)(17-18)-C (Xaa)(3-5)-H (HCCH) motif. The HCCH motif is involved in stabilizing the Vif Cullin 5 interaction, but the exact role of the conserved His and Cys residues remains elusive. In this report, we find that full-length HIV-1 Vif, as well as a HCCH peptide, is capable of binding to zinc with high specificity. Zinc binding induces a conformational change that leads to the formation of large protein aggregates. EDTA reversed aggregation and regenerated the apoprotein conformation. Cysteine modification studies with the HCCH peptide suggest that C114 is critical for stabilizing the fold of the apopeptide, and that C133 is located in a solvent-exposed region with no definite secondary structure. Selective alkylation of C133 reduced metal-binding specificity of the HCCH peptide, allowing cobalt to bind with rates comparable to that with zinc. This study demonstrates that the HCCH motif of HIV-1 Vif is a unique metal-binding domain capable of mediating protein-protein interactions in the presence of zinc and adds to a growing list of examples in which metal ion binding induces protein misfolding and/or aggregation. PMID- 17132732 TI - Metabolic syndrome and obesity in an insect. AB - Dragonflies infected with noninvasive gregarine gut parasites (Apicomplexa: Eugregarinorida) [corrected] have reduced flight-muscle performance, an inability to metabolize lipid in their muscles, twofold-elevated hemolymph carbohydrate concentrations, and they accumulate fat in their thorax in a manner analogous to mammalian obesity. Gregarine infection is associated with inappropriate responses of hemolymph carbohydrate concentration to insulin and with chronic activation in the flight muscles of p38 MAP kinase, a signaling molecule involved in immune and stress responses. Short-term exposure to gregarine excretory/secretory products caused elevated blood carbohydrate and p38 MAPK activation in healthy individuals. These characteristics comprise a set of symptoms and processes that are known in mammals as metabolic syndrome but which have not previously been described in other animal taxa. In addition to expanding the known taxonomic breadth of metabolic disease, these results indicate that insects may be useful experimental models for studying its underlying biology and mechanisms. PMID- 17132733 TI - Flexible bilayers with spontaneous curvature lead to lamellar gels and spontaneous vesicles. PMID- 17132734 TI - Vesicle transport, cilium formation, and membrane specialization: the origins of a sensory organelle. PMID- 17132735 TI - Distant touch hydrodynamic imaging with an artificial lateral line. AB - Nearly all underwater vehicles and surface ships today use sonar and vision for imaging and navigation. However, sonar and vision systems face various limitations, e.g., sonar blind zones, dark or murky environments, etc. Evolved over millions of years, fish use the lateral line, a distributed linear array of flow sensing organs, for underwater hydrodynamic imaging and information extraction. We demonstrate here a proof-of-concept artificial lateral line system. It enables a distant touch hydrodynamic imaging capability to critically augment sonar and vision systems. We show that the artificial lateral line can successfully perform dipole source localization and hydrodynamic wake detection. The development of the artificial lateral line is aimed at fundamentally enhancing human ability to detect, navigate, and survive in the underwater environment. PMID- 17132736 TI - Dendritic spines linearize the summation of excitatory potentials. AB - In mammalian cortex, most excitatory inputs occur on dendritic spines, avoiding dendritic shafts. Although spines biochemically isolate inputs, nonspiny neurons can also implement biochemical compartmentalization; so, it is possible that spines have an additional function. We have recently shown that the spine neck can filter membrane potentials going into and out of the spine. To investigate the potential function of this electrical filtering, we used two-photon uncaging of glutamate and compared the integration of electrical signals in spines vs. dendritic shafts from basal dendrites of mouse layer 5 pyramidal neurons. Uncaging potentials onto spines summed linearly, whereas potentials on dendritic shafts reduced each other's effect. Linear integration of spines was maintained regardless of the amplitude of the response, distance between spines (as close as < 2 microm), distance of the spines to the soma, dendritic diameter, or spine neck length. Our findings indicate that spines serve as electrical isolators to prevent input interaction, and thus generate a linear arithmetic of excitatory inputs. Linear integration could be an essential feature of cortical and other spine-laden circuits. PMID- 17132737 TI - Attenuation of signaling pathways stimulated by pathologically activated FGF receptor 2 mutants prevents craniosynostosis. AB - Craniosynostosis, the fusion of one or more of the sutures of the skull vault before the brain completes its growth, is a common (1 in 2,500 births) craniofacial abnormality, approximately 20% of which occurrences are caused by gain-of-function mutations in FGF receptors (FGFRs). We describe a genetic and pharmacological approach for the treatment of a murine model system of Crouzon like craniosynostosis induced by a dominant mutation in Fgfr2c. Using genetically modified mice, we demonstrate that premature fusion of sutures mediated by Crouzon-like activated Fgfr2c mutant is prevented by attenuation of signaling pathways by selective uncoupling between the docking protein Frs2alpha and activated Fgfr2c, resulting in normal skull development. We also demonstrate that attenuation of Fgfr signaling in a calvaria organ culture with an Fgfr inhibitor prevents premature fusion of sutures without adversely affecting calvaria development. These experiments show that attenuation of FGFR signaling by pharmacological intervention could be applied for the treatment of craniosynostosis or other severe bone disorders caused by mutations in FGFRs that currently have no treatment. PMID- 17132738 TI - Prediction of active nodes in the transcriptional network of neural tube patterning. AB - A transcriptional network governs patterning in the developing spinal cord. As the developmental program runs, the levels of sequence-specific DNA-binding transcription factors (SSTFs) in each progenitor cell type change to ultimately define a set of postmitotic populations with combinatorial codes of expressed SSTFs. A network description of the neural tube (NT) transcriptional patterning process will require definition of nodes (SSTFs and target enhancers) and edges (interactions between nodes). There are 1,600 SSTF nodes in a given mammalian genome. To limit the complexity of a network description, it will be useful to discriminate between active and passive SSTF nodes. We define active SSTF nodes as those that are differentially expressed within the system. Our system, the developing NT, was partitioned into two pools of genetically defined populations by using flow sorting. Microarray comparisons across the partition led to an estimate of 500-700 active SSTF nodes in the transcriptional network of the developing NT. These included most of the 66 known SSTFs assembled from review articles and recent reports on NT patterning. Empirical cutoffs based on the performance of knowns were used to identify 188 further active SSTFs nodes that performed similarly. The general utility and limitations of the population partitioning paradigm are discussed. PMID- 17132739 TI - Topographical complexity of multidimensional energy landscapes. AB - A scheme for visualizing and quantifying the complexity of multidimensional energy landscapes and multiple pathways is presented employing principal component-based disconnectivity graphs and the Shannon entropy of relative "sizes" of superbasins. The principal component-based disconnectivity graphs incorporate a metric relationship between the stationary points of the system, which enable us to capture not only the actual assignment of the superbasins but also the size of each superbasin in the multidimensional configuration space. The landscape complexity measure quantifies the degree of topographical complexity of a multidimensional energy landscape and tells us at which energy regime branching of the main path becomes significant, making the system more likely to be kinetically trapped in local minima. The path complexity measure quantifies the difficulty encountered by the system to reach a connected local minimum by the path in question, implying that the more significant the branching points along the path the more difficult it is to end up in the desired local minimum. As an illustrative example, we apply this analysis to two kinds of small model protein systems exhibiting a highly frustrated and an ideal funnel-like energy landscape. PMID- 17132740 TI - Anthrax pathogen evades the mammalian immune system through stealth siderophore production. AB - Systemic anthrax, caused by inhalation or ingestion of Bacillus anthracis spores, is characterized by rapid microbial growth stages that require iron. Tightly bound and highly regulated in a mammalian host, iron is scarce during an infection. To scavenge iron from its environment, B. anthracis synthesizes by independent pathways two small molecules, the siderophores bacillibactin (BB) and petrobactin (PB). Despite the great efficiency of BB at chelating iron, PB may be the only siderophore necessary to ensure full virulence of the pathogen. In the present work, we show that BB is specifically bound by siderocalin, a recently discovered innate immune protein that is part of an antibacterial iron-depletion defense. In contrast, neither PB nor its ferric complex is bound by siderocalin. Although BB incorporates the common 2,3-dihydroxybenzoyl iron-chelating subunit, PB is novel in that it incorporates the very unusual 3,4-dihydroxybenzoyl chelating subunit. This structural variation results in a large change in the shape of both the iron complex and the free siderophore that precludes siderocalin binding, a stealthy evasion of the immune system. Our results indicate that the blockade of bacterial siderophore-mediated iron acquisition by siderocalin is not restricted to enteric pathogenic organisms and may be a general defense mechanism against several different bacterial species. Significantly, to evade this innate immune response, B. anthracis produces PB, which plays a key role in virulence of the organism. This analysis argues for antianthrax strategies targeting siderophore synthesis and uptake. PMID- 17132741 TI - Premature arrest of the male flower meristem precedes sexual dimorphism in the dioecious plant Silene latifolia. AB - Most dioecious plant species are believed to derive from hermaphrodite ancestors. The regulatory pathways that have been modified during evolution of the hermaphrodite ancestors and led to the emergence of dioecious species still remain unknown. Silene latifolia is a dioecious plant species harboring XY sex chromosomes. To identify the molecular mechanisms involved in female organ suppression in male flowers of S. latifolia, we looked for genes potentially involved in the establishment of floral organ and whorl boundaries. We identified homologs of Arabidopsis thaliana SHOOTMERISTEMLESS (STM) and CUP SHAPED COTYLEDON (CUC) 1 and CUC2 genes in S. latifolia. Our phylogenetic analyses suggest that we identified true orthologs for both types of genes. Detailed expression analyses showed a conserved expression pattern for these genes between S. latifolia and A. thaliana, suggesting a conserved function of the corresponding proteins. Comparative in situ hybridization experiments between male, female, and hermaphrodite individuals reveal that these genes show a male-specific pattern of expression before any morphological difference become apparent. Our results make SlSTM and SlCUC strong candidates for being involved in sex determination in S. latifolia. PMID- 17132742 TI - Silencing leaf sorbitol synthesis alters long-distance partitioning and apple fruit quality. AB - Sorbitol and sucrose are major products of photosynthesis distributed in apple trees (Malus domestica Borkh. cv. "Greensleeves") that affect quality in fruit. Transgenic apple plants were silenced or up-regulated for sorbitol-6-phosphate dehydrogenase by using the CaMV35S promoter to define the role of sorbitol distribution in fruit development. Transgenic plants with suppressed sorbitol-6 phosphate dehydrogenase compensated by accumulating sucrose and starch in leaves, and morning and midday net carbon assimilation rates were significantly lower. The sorbitol to sucrose ratio in leaves was reduced by approximately 90% and in phloem exudates by approximately 75%. The fruit accumulated more glucose and less fructose, starch, and malic acid, with no overall differences in weight and firmness. Sorbitol dehydrogenase activity was reduced in silenced fruit, but activities of neutral invertase, vacuolar invertase, cell wall-bound invertase, fructose kinase, and hexokinase were unaffected. Analyses of transcript levels and activity of enzymes involved in carbohydrate metabolism throughout fruit development revealed significant differences in pathways related to sorbitol transport and breakdown. Together, these results suggest that sorbitol distribution plays a key role in fruit carbon metabolism and affects quality attributes such as sugar-acid balance and starch accumulation. PMID- 17132743 TI - West Nile virus nonstructural protein NS1 inhibits complement activation by binding the regulatory protein factor H. AB - The complement system, by virtue of its dual effector and priming functions, is a major host defense against pathogens. Flavivirus nonstructural protein (NS)-1 has been speculated to have immune evasion activity, because it is a secreted glycoprotein, binds back to cell surfaces, and accumulates to high levels in the serum of infected patients. Herein, we demonstrate an immunomodulatory function of West Nile virus NS1. Soluble and cell-surface-associated NS1 binds to and recruits the complement regulatory protein factor H, resulting in decreased complement activation in solution and attenuated deposition of C3 fragments and C5b-9 membrane attack complexes on cell surfaces. Accordingly, extracellular NS1 may function to minimize immune system targeting of West Nile virus by decreasing complement recognition of infected cells. PMID- 17132744 TI - Testosterone and cognitive function: current clinical evidence of a relationship. AB - BACKGROUND: Testosterone levels decline as men age, as does cognitive function. Whether there is more than a temporal relationship between testosterone and cognitive function is unclear. Chemical castration studies in men with prostate cancer suggest that low serum testosterone may be associated with cognitive dysfunction. Low testosterone levels have also been observed in patients with Alzheimer's disease (AD) and mild cognitive impairment (MCI). This paper reviews the current clinical evidence of the relationship between serum testosterone levels and cognitive function in older men. METHODS: A systematic literature search was conducted using PubMed and EMBASE to identify clinical studies and relevant reviews that evaluated cognitive function and endogenous testosterone levels or the effects of testosterone substitution in older men. RESULTS: Low levels of endogenous testosterone in healthy older men may be associated with poor performance on at least some cognitive tests. The results of randomized, placebo-controlled studies have been mixed, but generally indicate that testosterone substitution may have moderate positive effects on selective cognitive domains (e.g. spatial ability) in older men with and without hypogonadism. Similar results have been found in studies in patients with existing AD or MCI. CONCLUSIONS: Low endogenous levels of testosterone may be related to reduced cognitive ability, and testosterone substitution may improve some aspects of cognitive ability. Measurement of serum testosterone should be considered in older men with cognitive dysfunction. For men with both cognitive impairment and low testosterone, testosterone substitution may be considered. Large, long-term studies evaluating the effects of testosterone substitution on cognitive function in older men are warranted. PMID- 17132745 TI - Remission of Graves' disease during anti-thyroid drug therapy. Time to reconsider the mechanism? AB - Therapy of Graves' hyperthyroidism with thionamide anti-thyroid drugs is accompanied by a gradual remission of the autoimmune aberration in the majority of patients. The most likely mechanism behind this remission has been considered to be a direct immunosuppressive effect of thionamide drugs. However, a number of findings in clinical studies of patients with Graves' disease indicate that remission is probably not caused by a special effect of thionamide drugs. Many studies have shown that remission is linked to restoration of the euthyroid state, and that it is independent of drug dose and type. Moreover, similar remission is observed when patients become euthyroid after thyroid surgery. In an explanatory model described, it is assumed that the autoimmune aberration of Graves' disease is often basically quit mild and self-limiting. Patients may become ill by the running of a vicious cycle of hyperthyroidism worsening the autoimmunity, and autoimmunity worsening the hyperthyroidism. Once patients are made euthyroid by one or the other drug or by thyroid surgery, the majority of patients will gradually enter remission of the disease. The conclusion that remission is associated with restoration of the euthyroid state, and that it is not a special drug effect, highlights the importance of making and keeping patients with Graves' disease euthyroid. PMID- 17132746 TI - Thiamine transporter mutation: an example of monogenic diabetes mellitus. AB - OBJECTIVE: Thiamine-responsive megaloblastic anemia (TRMA) is a rare syndrome characterized by diabetes mellitus (DM), anemia, and sensorineural deafness. We describe the clinical course and the molecular defect of a young woman who was diagnosed to have this syndrome. CASE: The patient is an 18-year-old girl who was born to non-consanguous parents. She was noted to be deaf-mute in the first year of life. She was diagnosed with DM at the age of 9 months and with severe anemia at the age of 2 years. An extensive work up could not identify the cause. She was treated with blood transfusions every 3-4 weeks for the past 16 years. A diagnosis of TRMA was suspected and the patient was treated with thiamine hydrochloride. Hemoglobin and platelets increased to normal values after a few weeks of thiamine therapy. Diabetic control significantly improved but she had no noticeable changes in the deafness. METHODS: Peripheral blood DNA was extracted from the patient, her mother, aunt, and a healthy sister. Exons and exon-intron boundaries of the thiamine transporter gene SLC19A2 were PCR amplified and directly sequenced. RESULTS: A G515C homozygous mutation was identified in the SLC19A2 gene of the patient. This mutation changes Gly to Arg at codon 172 (G172R). The mother, an aunt, and a sister had a heterozygous G172R mutation. CONCLUSIONS: Mutations in thiamine transporter gene, SLC19A2, causes a rare form of monogenic diabetes, anemia, and sensorineural deafness. Thiamine induces a remarkable hematological response and improvement in the diabetic control but has no effect on deafness. PMID- 17132747 TI - Evolution of gonadotropin deficiency in a patient with type II autosomal dominant GH deficiency. AB - BACKGROUND: Type II isolated GH deficiency (IGHD type II) is caused by dominant negative splicing or point mutations of the GH-1 gene. Studies have suggested that dominant mutant GH forms prevent the secretion of wild-type GH, resulting in eventual cell death; surprisingly, some patients with these GH mutations develop other hormonal deficiencies (ACTH, TSH). SUBJECTS: The proband presented at the age of 2.3 years with IGHD. His father, also known to have been treated for IGHD as a child, had subsequently been lost to follow-up, having remained without treatment during this time. At re-evaluation at the age of 38 years, he complained of lack of stamina and poor libido. Clinical and biochemical assessment confirmed severe GHD, borderline ACTH insufficiency, suboptimal basal and stimulated gonadotropins, and a poor prolactin response to provocation. The basal testosterone concentration was low, and he complained of secondary infertility. Magnetic resonance imaging revealed anterior pituitary hypoplasia in both patients. Genetic testing revealed a heterozygous splicing mutation in GH-1 (intervening sequence-3 + 1G>A) in both patients, known to cause IGHD type II. INTERVENTIONS: The proband showed an excellent growth response to recombinant human GH (rhGH). His father, also treated with rhGH, showed improved quality of life on rhGH, but testosterone concentrations continued to decline, necessitating treatment with testosterone with symptomatic benefit but no improvement in semen quality. CONCLUSIONS: This case supports recent experimental and clinical observations suggesting that the cytotoxicity associated with accumulation of dominant negative mutant 17.5 kDa GH causes a form of GHD that can evolve into multiple hormone deficiencies. Hence, patients diagnosed initially with IGHD type II require continued long-term clinical follow-up. PMID- 17132748 TI - The incidence of second primary tumors in thyroid cancer patients is increased, but not related to treatment of thyroid cancer. AB - OBJECTIVE: The aim of the present study is to assess the prevalence of second primary tumors in patients treated for thyroid cancer. Furthermore, we wanted to assess the standardized risk rates for all second primary tumors, but especially for breast cancer, as data in the literature indicate an excessive risk in differentiated thyroid cancer (DTC) patients for this tumor. MATERIALS AND METHODS: We included consecutive patients, who received ablation treatment with I 131 at the Leiden University Medical Center between January 1985 and December 1999 (n = 282). The mean period of follow-up was 10.6 +/- 4.1 years. RESULTS: Thirty-five of the 282 patients (12.4%) had a second primary tumor (SPT), either preceding or following the diagnosis of thyroid cancer. Five other patients had three primary tumors, including DTC. As a result, 40 additional tumors were found in this group, revealing an overall prevalence of 14.2%. Twenty tumors (7.1%) preceded the thyroid cancer with a mean interval of 5.7 years (range: 0.5-22.0 years), whereas 20 tumors (7.1%) occurred after this tumor with a mean interval of 6.7 years (range: 1.0-15.0 years). In 13 female patients, breast cancer was found as SPT. The standardized incidence rate (SIR) for all cancers after the diagnosis of DTC in this study population was not increased (1.13; confidence interval (CI): 0.68-1.69). However, we found an increased SIR of 2.26 (CI: 1.60 3.03) for all cancers either following or preceding DTC, which is mainly caused by a SIR of 3.95 (CI: 2.06-6.45) for breast cancer. CONCLUSION: Patients with DTC have an overall increased standardized incidence rate for second primary tumors, but not for second primary tumors following I-131 therapy. These findings suggest a common etiologic and/or genetic mechanism instead of a causal relation. PMID- 17132749 TI - Selenium and goiter prevalence in borderline iodine sufficiency. AB - DESIGN: Selenium (Se) is required for the biosynthesis of selenocysteine containing proteins. Several selenoenzymes, e.g. glutathione peroxidases and thioredoxin reductases, are expressed in the thyroid. Selenoenzymes of the deiodinase family regulate the levels of thyroid hormones. For clinical investigators, it is difficult to determine the role of Se in the etiology of (nodular-)goiter, because there are considerable variations of Se concentrations in different populations as reflected by dietary habits, bioavailability of Se compounds, and racial differences. Moreover, most previous clinical trials which investigated the influence of Se on thyroid volume harbored a bias due to the coexistence of severe iodine deficiency in the study populations. METHODS: Therefore, we investigated the influence of Se on thyroid volume in an area with borderline iodine sufficiency. First, we investigated randomly selected probands for urinary iodine (UI) and creatinine excretion in spot urine samples and determined the prevalence of goiter and thyroid nodules by high-resolution ultrasonography. After this, we determined urinary Se excretion (USe) in probands with goiter as well as in matched probands without goiter. Adjustments between the two compared groups were made for age, gender, history of thyroid disorders, smoking, and UI excretion. RESULTS: The mean USe and UI rates of all 172 probands were 24 micro g Se/l or 27 micro g Se/g creatinine and 96 micro g I/l or 113 micro g I/g creatinine indicating borderline selenium (20-200 micro g/l) and iodine (100-200 micro g/l) sufficiency of the study population. Probands with goiter (n=89) showed significantly higher USe levels than probands with normal thyroid volume (n=83; P < 0.05). USe rates were not influenced by present smoking or pregnancy. CONCLUSIONS: In our investigation, USe was not an independent risk factor for the development of goiter. The higher USe in probands with goiter in comparison with probands with normal thyroid volume is most likely a coincidence. Se does not significantly influence thyroid volume in borderline iodine sufficiency because the iodine status is most likely the more important determinant. PMID- 17132750 TI - Selective parathyroid venous sampling in patients with complicated hyperparathyroidism. AB - OBJECTIVE: The role of preoperative localisation of abnormal parathyroid glands remains controversial but is particularly relevant to the management of patients with recurrent or persistent hyperparathyroidism and familial syndromes. We report our experience of the use of selective parathyroid venous sampling (PVS) in the localisation of parathyroid disease in such patients. DESIGN: We report a retrospective 10-year experience (n = 27) of the use of PVS in complicated primary hyperparathyroidism and contrast the use of PVS with neck ultrasound, magnetic resonance imaging (MRI), computed tomography (CT) and sestamibi imaging modalities. RESULTS: In 14 out of 25 patients who underwent surgery PVS results were completely concordant with surgical and histological findings and 88% of patients achieved post-operative cure. Out of 13 patients referred after previous failed surgery, 12 underwent further surgery which was curative in 9. In total PVS yielded useful positive (n = 13) and/or negative information (n = 6) in 19 out of 25 patients undergoing surgery. Using histology as the gold standard, 59% of PVS studies were entirely consistent with histology, as compared with 39% of ultrasound scans, 36% of sestamibi scans and 17% of MRI/CT scans. CONCLUSIONS: PVS is a valuable adjunct to MRI/CT and sestamibi scanning in selected patients with complicated hyperparathyroidism when performed in an experienced unit. PMID- 17132751 TI - Non-functioning pituitary adenoma database: a useful resource to improve the clinical management of pituitary tumors. AB - OBJECTIVE: The long-term outcome of non-functioning pituitary adenoma (NFPA) patients is not clearly established, probably due to the low annual incidence and prolonged natural history of these rare tumors. The aim of this study was to evaluate clinical data at presentation and long-term post-surgery and radiotherapy outcome in a cohort of patients with NFPA. DESIGN AND METHODS: A computerized database was developed using Access 2000 software (Microsoft Corporation, 1999). Retrospective registration of 295 NFPA patients was performed in seven Endocrinological Centers of North West Italy. Data were analyzed by STATA software. RESULTS: The main presenting symptoms were visual defects (67.8%) and headache (41.4%) and the most frequent pituitary deficit was hypogonadism (43.3%), since almost all tumors were macroadenomas (96.5%). Surgery was the first choice treatment (98% of patients) and total debulking was achieved in 35.5%. Radiotherapy was performed as adjuvant therapy after surgery in 41% of patients. At the follow-up, recurrence occurred in 19.2% of patients without post surgical residual tumor after 7.5 +/- 2.6 years, regrowth in 58.4% of patients with post-surgical remnant after 5.3 +/- 4.0 years and residue enlargement in 18.4% of patients post-surgically treated with radiotherapy after 8.1 +/- 7.3 years. CONCLUSIONS: Our database indicates that the goal of a definitive surgical cure has been achieved during the last decade in a low percentage of patients with NFPA. This tumor database may help to reduce the delay between symptom onset and diagnosis, to assess prognostic parameters for the follow-up of patients with different risk of recurrence and to define the efficacy and safety of different treatments and their association with mortality/morbidity. PMID- 17132752 TI - Associations of remission status and lanreotide treatment with quality of life in patients with treated acromegaly. AB - OBJECTIVE: Acromegaly has an important impact on quality of life (QOL). The aim of this study was to evaluate the associations of remission status and lanreotide treatment with QOL in patients with treated acromegaly, by the newly developed disease-generated acromegaly QOL questionnaire (ACROQOL). DESIGN: Cross-sectional study. METHODS: Fifty-two patients with treated acromegaly were recruited to complete the Chinese version of the ACROQOL translated and validated from the English version. These patients were divided into controlled and uncontrolled groups based on the latest remission criteria and further subdivided into four groups according to the present treatment with lanreotide or not. Comparisons between groups were analyzed. RESULTS: There was no difference between controlled and uncontrolled groups in the ACROQOL scores of total score, both scales and psychological subscales. However, in the controlled group, present treatment with lanreotide, in comparison with no treatment, showed worse ACROQOL scores in total score (P = 0.021), psychological scale (P = 0.011), psychological subscale 'appearance' (P = 0.032) and 'personal relations' (P = 0.010). CONCLUSIONS: The lanreotide treatment was negatively associated with QOL in biochemically controlled acromegalic patients, especially in the psychological aspect. PMID- 17132753 TI - Searching for somatic mutations in McCune-Albright syndrome: a comparative study of the peptidic nucleic acid versus the nested PCR method based on 148 DNA samples. AB - BACKGROUND: Activating mutations of the Gsalpha gene (GNAS), which encodes for the alpha-subunit of the stimulatory G protein, have been identified in patients with McCune-Albright syndrome (MAS). Accuracy and sensitivity in the molecular diagnosis of MAS is mandatory for optimal therapeutic strategy and adapted follow up, especially for incomplete clinical forms of MAS. To date, the highly sensitive nested PCR method with intermediary digestion by a restriction enzyme at the mutation site is one of the most widely used techniques. This study evaluated a new diagnostic method using a peptidic nucleic acid (PNA) and compared it with the nested PCR method. MATERIAL AND METHODS: One hundred and forty-eight DNA samples from eighty-eight patients presenting clinical symptoms compatible with MAS were included. The DNA samples were mainly obtained from peripheral blood, ovarian tissue or cyst liquid, and bone lesions. The nested PCR method required 4 days. PNA clamping required 1.5 days and utilized the higher thermal stability and specificity of PNA-DNA coupling to inhibit PCR product formation. Direct sequencing was subsequently performed in all cases. RESULTS: The sensitivity of mutation detection was 54% (n = 80) for nested PCR and 46.6% (n = 69) for PNA (P > 0.05). The 11 cases where PNA failed to detect the mutation were mainly incomplete and atypical clinical forms of MAS (n = 10/11). The cost per sample was 50 Euros for PNA clamping versus 136 Euros for nested PCR. CONCLUSION: PNA clamping is a rapid, reliable, and economical method to diagnose MAS. It should be the first-line diagnostic method, although negative results, especially for incomplete clinical forms of MAS, should be confirmed by nested PCR. PMID- 17132754 TI - Effect of exogenous glucocorticoid on osmotically stimulated antidiuretic hormone secretion and on water reabsorption in man. AB - OBJECTIVE: Glucocorticoids exert tonic suppression of antidiuretic hormone (ADH) secretion. Hypocortisolism in secondary adrenocortical insufficiency can result in a clinical picture similar to the syndrome of inappropriate ADH secretion. On the other hand, in vitro and in vivo results provide evidence for ADH suppression in states of hypercortisolism. To test the hypothesis that ADH suppression is of relevance during glucocorticoid therapy, we investigated the influence of prednisolone on the osmotic stimulation of ADH. DESIGN AND METHODS: Seven healthy men were subjected to water deprivation tests with the measurement of plasma ADH (pADH) and osmolality (posmol) before and after glucocorticoid treatment (5 days 30 mg prednisolone per day). RESULTS: Before glucocorticoid treatment, the volunteers showed a normal test with an adequate increase of pADH (basal 0.54 +/- 0.2 to 1.9 +/- 0.72 pg/ml (mean +/- S.D.)) in relation to posmol(basal 283.3 +/- 8.5 to 293.7 +/- 6 mosmol/kg). After prednisolone intake, pADH was attenuated (<0.4 pg/ml) in spite of an increase of posmol from 289.3 +/- 3.6 to 297.0 +/- 5.5 mosmol/kg. However, urine osmolar concentration increased normally during water deprivation after prednisolone. Urinary cAMP excretion increased during water deprivation without glucocorticoid treatment from 3.56 +/- 0.55 to 6.07 +/- 0.76 micro mol/l, reflecting the increased pADH levels. The rise in cAMP excretion was completely blunted by prednisolone treatment. CONCLUSIONS: We speculate that there may be an ADH-independent stimulation of the formation or function of aquaporin-2 channels by prednisolone and/or a direct osmotic stimulation of water reabsorption independent of ADH and glucocorticoid control. PMID- 17132755 TI - The analysis of quantitative expression of somatostatin and dopamine receptors in gastro-entero-pancreatic tumours opens new therapeutic strategies. AB - OBJECTIVE: Somatostatin (sst) are present in the majority of gastro-entero pancreatic (GEP) tumours. Effects of somatostatin receptor (sst) analogues are partial and of limited duration. Cell lines derived from GEP express dopaminergic receptors D(2). New chimeric analogues simultaneously recognising sst(2) and sst(5) or sst(2) and D(2) have additive effects in inhibition of GH and prolactin secretion in pituitary adenomas. Our aim was to quantify the expression of sst and D(2) mRNA in human GEP tumours. DESIGN AND METHODS: mRNA expression of sst(1), sst(2), sst(3) and sst(5) as well as D(2), was analysed using real-time PCR (TaqMan probe) in a series of 35 patients with GEP tumours (pancreas (n = 19) and intestinal (n = 16)). Levels of expression were compared with a group of 13 somatotroph adenomas. RESULTS: All GEP tumours express sst(1), sst(2) and D(2). Expression of sst(3) and sst(5) was observed in 89 and 76% of tumours respectively with highly variable levels. sst(2) mRNA expression was higher in nonfunctional tumours (P < 0.009) and sst5 was higher in pancreatic than in intestinal tumours (P < 0.02). Whereas sst(2) levels were similar between GEP and somatotroph tumours, levels of sst(5) and D(2) were higher in the former (394.9 +/- 156.1 x 10(-2) vs 69.7 +/- 19.5 x 10(-2) copy/copy beta-Gus (P < 0.0036) and 519.6 +/- 121.2 x 10(-2) vs 50.0 +/- 21.6 x 10(-2) copy/copy beta-Gus (P < 0.0001) respectively). In small tumours ( < 30 mm), sst(2) density appeared as a crucial parameter in somatostatin receptor scintigraphy results, whereas in big tumours, a consistent bias in SRS results was introduced by the size. In pancreatic GEP, high-level sst(3) expression was found in tumours with more active angiogenesis (higher microvessel density and vascular endothelial growth factor expression (P < 0.03)). CONCLUSIONS: GEP tumours co-express sst(2) and D(2) in 100% of cases and sst(5) in 89% thus supporting the testing of bi specific agonists (sst(2)/sst(5) or sst(2)/D(2)) in these tumours. PMID- 17132756 TI - Lower insulin sensitivity differentiates hirsute from non-hirsute Sicilian women with polycystic ovary syndrome. AB - OBJECTIVE: It is well known that hyperandrogenism and insulin-resistance with or without compensatory hyperinsulinism are closely associated, but the Rotterdam Consensus has concluded that principally obese women with polycystic ovary syndrome (PCOS) should be evaluated for the metabolic syndrome. Our aim was to study insulin sensitivity in PCOS women with hirsutism regardless of obesity. METHODS: Clinical characteristics, sex hormones and fasting- and after OGTT glycemia and insulinemia, homeostatic model of insulin resistance (HOMA IR), and Matsuda index of insulin sensitivity were analyzed in 130 women with PCOS. Hirsutism has been evaluated through the Ferriman-Gallwey (FG) map scoring system. RESULTS: PCOS women with hirsutism (57.7% of participants) showed significant higher values of total testosterone levels (P = 0.016), free testosterone (P = 0.027), DHEA sulfate (P = 0.017), and Delta4androstenedione (P = 0.018). They had similar body mass index (BMI) (P = 0.073) and were significantly less insulin sensitive (P = 0.002) than those without hirsutism (42.3% of participants). In women with PCOS and hirsutism, there was a significant correlation between FG score and insulin-sensitivity indexes (HOMA IR, rho = 0.33, P = 0.005; Matsuda index, rho = -0.34, P = 0.003) but not with the androgen levels. Moreover, women with hirsutism showed a significantly greater insulin (P = 0.019), C-peptide (P = 0.002), and glucose (P = 0.024) areas under the curve (auc2h). CONCLUSIONS: Our study suggests that the increased responsiveness of the pilo-sebaceous unit to androgens seems to be influenced by insulin sensitivity and that insulin resistance should be assessed in all hirsute women with PCOS regardless of their BMI, as insulin resistance was found in hirsute women irrespective of whether they were overweight or obese. PMID- 17132757 TI - Effects of modest testosterone supplementation and exercise for 12 weeks on body composition and quality of life in elderly men. AB - OBJECTIVE: One of the factors that may promote deterioration in quality of life and body composition in elderly men is the relative decline in serum testosterone levels with aging. In this study, we assessed the effects of modest doses of testosterone and a home-based strengthening program on quality of life and body composition in elderly men with relative testosterone insufficiency. DESIGN: Double-blind, placebo-controlled randomized study (testosterone), and additional randomization to a resistance exercise program or no additional exercise for 12 weeks in men between ages of 65 and 85 years with relative testosterone insufficiency. METHODS: Seventy sedentary, community dwelling men were randomized to a 5 mg testoderm transdermal system applied daily vs placebo system, and additionally randomized to a home-based resistance exercise program. Subjects were randomized to Group 1 (testosterone plus exercise), Group 2 (testosterone plus no exercise), Group 3 (placebo plus exercise), and Group 4 (placebo plus no exercise). Endpoints included quality of life (assessed by the short form-36 questionnaire) and body composition (measured by dual x-ray absorptiometry scan). RESULTS: Serum testosterone increased by a mean of 10.0 +/- 1.9, 6.6 +/- 1.6, 0.52 +/- 0.6, and 0.5 +/- 0.6 nmol/l in Groups 1, 2, 3, and 4 respectively. There was a significant interaction of testosterone and exercise on quality of life in the domains of physical functioning (P = 0.03), role physical (P = 0.01), general health (P = 0.049), and social functioning (P = 0.04). There were no effects of testosterone or exercise on quality of life alone, nor in body composition parameters. CONCLUSIONS: Modest testosterone supplementation to elderly men with relative testosterone insufficiency improved quality of life when accompanied by an exercise program. The combination of testosterone and exercise may be an important strategy in the elderly, though further studies are necessary to determine the long-term impact on body composition and function and for analysis of risk/benefit ratios as well. PMID- 17132758 TI - Quality of life in 70 women with disorders of sex development. AB - OBJECTIVE: The aim of this study was to assess the quality of life and psychosocial well-being in women with disorders of sex development (DSD). DESIGN: An open case-control study. METHODS: Social and psychiatric information was collected via a structured interview from 70 Danish women diagnosed with DSD, 70 controls matched on sex, age, and school education, and six women with isolated genital malformations. Quality of life and mental distress were assessed by 'Quality of Life-Assessment of Growth Hormone Deficiency in Adults' (QoL-AGHDA) and three symptom scales from the 'Hopkins Symptom Checklist' (SCL-90-R; i.e. somatization, depression, and anxiety) respectively. For both measures, higher scores reflected poorer outcomes. RESULTS: Present relationships and having children were less frequent in patients than in controls (P = 0.02 and P < 0.001 respectively). Previous suicidal thoughts (P = 0.002) and a higher frequency of psychological/psychiatric counseling for severe problems (P = 0.06) were more frequently reported in patients than in controls. The mean QoL-AGHDA score was significantly higher in patients than in controls (5.5 vs 2.9; P = 0.002), especially for congenital adrenal hyperplasia (CAH) females (P = 0.01) and virilized 46,XX and 46,XY females (P = 0.04). The total SCL score was higher in patients than in controls (mean 23.2 vs 20.0), reaching significance for anxiety (mean 6.3 vs 4.3, P = 0.03) with highest score in CAH (P = 0.01). CONCLUSION: An impaired quality of life and more affective distress were observed especially in CAH patients and virilized 46,XX and 46,XY females. This may be caused by trauma from distressing diagnostic procedures, the chronic illnesses per se, and psychosocial consequences of the disorders. PMID- 17132759 TI - Serum sex hormone and plasma homocysteine levels in middle-aged and elderly men. AB - OBJECTIVE: To investigate whether circulating levels of testosterone (total, bioavailable), estradiol (total, bioavailable), and DHEA sulfate (DHEAS) are associated with fasting plasma homocysteine (tHcy) levels in middle-aged and elderly men. DESIGN: A population-based sample of 400 independently living men between 40 and 80 years of age in a cross-sectional study. METHODS: Total testosterone, sex hormone binding globulin (SHBG), and total estradiol were measured by RIA methods and bioavailable testosterone and estradiol were calculated. DHEAS was measured using an immunometric technique. Fasting homocysteine was measured by fluorescence polarization immunoassay. Anthropometric characteristics were also measured and two standardized questionnaires completed, including life-style factors and diet. Linear regression analysis adjusted for age, body mass index (BMI), creatinine clearance, and mean visceral fat was used to assess the association of endogenous sex hormones and fasting plasma homocysteine levels. RESULTS: After adjustment for age, BMI, creatinine clearance, and mean visceral fat no statistically significant association was observed between testosterone (total, bioavailable), DHEAS, and estradiol (total, bioavailable)levels with natural log tHcy (beta = -2 x 10(-3); 95% confidence intervals (CI) -9 x 10(-3); 5 x 10(-3)), (beta = -4 x 10(-3); 95% CI -18 x 10(-3); 9 x 10(-3)), (beta = 3 x 10(-3); 95% CI -6 x 10(-3); 12 x 10(-3)), (beta = -9.3 x 10(-5); 95% CI -1 x 10(-3); 1 x 10(-3)), and (beta = 0.00; 95% CI -3 x 10(-3); 2 x 10(-3)) respectively. Additional adjustment for smoking, alcohol intake, daily physical activity, diabetes mellitus, and hypertension did not change these findings. CONCLUSION: The results of our study do not support a direct role for circulating sex hormone levels in the regulation of fasting plasma tHcy concentrations in middle-aged and elderly men. PMID- 17132760 TI - The Relationship between PON1 phenotype and PON1-192 genotype in detoxification of three oxons by human liver. AB - Phosphorothioate pesticides (OP) such as diazinon, chlorpyrifos, and parathion are activated to highly toxic oxon metabolites by the cytochromes P450 (P450s), mainly in the liver. Simultaneously, the P450s catalyze detoxification of OP to nontoxic dearylated metabolites. The oxon is then detoxified to the dearylated metabolite by PON1, an A-esterase present in the liver and blood serum. The aims of this study were to define the influence of PON1-192 genotype and phenotype on the capacity of human liver microsomes (n = 27) to detoxify the oxons diazoxon, chlorpyrifos-oxon, and paraoxon. Near physiological assay conditions were used to reflect as closely as possible metabolism in vivo and because the hydrolytic activity of the allelic variants of PON1-192 are differentially affected by a number of conditions. The rates of hydrolysis of diazoxon, chlorpyrifos-oxon, and paraoxon varied 5.7-, 16-, and 56-fold, respectively, regardless of PON1-192 genotype. Individuals with the PON1-192RR genotype preferentially hydrolyzed paraoxon (p < 0.01), and the R allele was associated with higher hydrolytic activity toward chlorpyrifos-oxon, but not diazoxon. There were strongly significant relationships between phenylacetate and paraoxon hydrolysis (p < 0.001) and phenylacetate and chlorpyrifos-oxon hydrolysis (p < 0.001), but not between phenylacetate and diazoxon hydrolysis. These data highlight the importance of PON1 phenotype for efficient hydrolysis of paraoxon and chlorpyrifos-oxon, but environmental and yet unknown genetic factors are more important than PON1-192 genotype in determining capacity to hydrolyze diazoxon. PMID- 17132761 TI - Pharmacokinetics, tissue distribution, metabolism, and excretion of depside salts from Salvia miltiorrhiza in rats. AB - Salviae miltiorrhiza, a traditional Chinese medical herb known as "Danshen," has been widely used in clinics to improve blood circulation, relieve blood stasis, and treat coronary heart disease. Depside salts from S. miltiorrhiza are a novel drug in which magnesium lithospermate B and its analogs are the active components. The pharmacokinetics, tissue distribution, metabolism, and excretion of three of the major components, lithospermic acid B, rosmarinic acid (RA), and lithospermic acid (LA), were studied by liquid chromatography-tandem mass spectrometry following intravenous administration in Sprague-Dawley rats. The elimination half-lives for LSB, RA, and LA were 1.04, 0.75, and 2.0 h, respectively, when 60 mg/kg S. miltiorrhiza depside salts were administrated. The areas under the curve for LSB, RA, and LA were 51.6, 6.6, and 25.2 mg . h/l, respectively, and the values decreased in the individual tissues in the following order: kidney > lung > liver > heart > spleen > brain for LSB; kidney > lung > heart > liver > spleen > brain for RA; and heart > lung > kidney > liver > spleen > brain for LA. After intravenous administration of 60 mg/kg S. miltiorrhiza depside salts, 86% of the LSB was excreted in the bile within 6 h. The main metabolites M1 and M2 were found in the serum. Overall, the results show that depside salts from S. miltiorrhiza are rapidly and widely distributed to tissues after intravenous administration in rats but that they are also rapidly cleared and excreted. PMID- 17132762 TI - Induction of glutathione synthesis explains pharmacodynamics of high-dose busulfan in mice and highlights putative mechanisms of drug interaction. AB - Busulfan is an example of a drug eliminated through glutathione S-transferase (GST)-catalyzed conjugation with reduced glutathione (GSH). We studied the pharmacokinetics and toxicity of busulfan in C57BL6 mice in correlation with liver GST activity and GSH synthesis by accurate determination of precursors, namely, gamma-glutamyl-cysteine and cysteine. A significantly lower incidence of acute toxicity was observed in mice receiving busulfan 16.5 mg/kg twice a day compared with animals receiving 33 mg/kg once a day. In both cases, a total dose of 132 mg/kg was administered over 4 days. The difference in toxicity was explained by pharmacokinetics since a strong induction of clearance was observed only in animals treated twice daily. Induction of metabolism was correlated with an increase in liver cysteine content and enhanced glutathione synthesis rate, whereas GST activity was unchanged. To our knowledge, this is the first time that in vivo flux of GSH synthesis has been shown to be closely related to a drug plasma clearance and toxicity. These results allow hypothesizing that GSH liver synthesis may directly influence busulfan clearance in humans with possible implications in the occurrence of hepatic veno-occlusive disease. PMID- 17132763 TI - Effects of pomegranate juice on human cytochrome P450 2C9 and tolbutamide pharmacokinetics in rats. AB - In this study, we investigated whether pomegranate juice could inhibit CYP2C9 activity. The ability of pomegranate juice to inhibit the diclofenac 4' hydroxylase activity of human CYP2C9 was examined using human liver microsomes. Pomegranate juice was shown to be a potent inhibitor of human CYP2C9. The addition of 25 microl (5% v/v) of pomegranate juice resulted in almost complete inhibition of human CYP2C9 activity. In addition, we investigated the effect of pomegranate juice on the pharmacokinetics of tolbutamide (substrate for CYP2C9) in rats. Relative to the control group, the area under the concentration-time curve was approximately 1.2-fold greater when pomegranate juice (3 ml) was injected p.o. 1 h before the p.o. administration of the tolbutamide (20 mg/kg). The elimination half-life of tolbutamide was not altered by pomegranate juice administration. These results suggest pomegranate juice ingestion inhibits the intestinal metabolism of tolbutamide without inhibiting the hepatic metabolism in rats. Thus, we discovered that pomegranate juice inhibited human CYP2C9 activity and furthermore increased tolbutamide bioavailability in rats. PMID- 17132764 TI - Evaluation of cryopreserved human hepatocytes as an alternative in vitro system to microsomes for the prediction of metabolic clearance. AB - Human liver microsomes have typically resulted in marked underprediction of in vivo human intrinsic clearance (CL(int)); therefore, the utility of cryopreserved hepatocytes as an alternative in vitro system has become an important issue. In this study, 10 compounds (tolbutamide, diclofenac, S-warfarin, S-mephenytoin, dextromethorphan, bufuralol, quinidine, nifedipine, testosterone, and terfenadine) were selected as substrate probes for CYP2C9, 2C19, 2D6, and 3A4, and the kinetics of metabolite formation (n = 14 pathways) were investigated in three individual lots of cryopreserved hepatocytes and in a pool of human liver microsomes. For the majority of the compounds, lower unbound K(M) or S(50) values were observed in hepatocytes compared with microsomes, on average by 50% over a 200-fold range (0.5-140 microM). Expressed on an equivalent liver weight basis, a good correlation between microsomal and hepatocyte V(max) values was observed for most pathways greater than 5 orders of magnitude (0.16-216 nmol/min/g liver). Unbound hepatocyte CL(int) (CL(int,u)) values, when scaled to the whole liver (range 0.38-4000 ml/min/kg), were on average 2.5-fold higher than microsomal CL(int,u) values, with the exception of tolbutamide and diclofenac, for which lower hepatocellular CL(int,u) values were observed. Hepatocyte predicted CL(int) values were compared with human in vivo CL(int) values, and to supplement our data, in vitro data from cryopreserved hepatocytes were collated from four other published sources. These data show that for 37 drugs, there is, on average, a 4.5 fold under-prediction of the in vivo CL(int) using cryopreserved hepatocytes, representing a significant reduction in prediction bias compared with human microsomes. PMID- 17132765 TI - Metabolism of MK-0524, a prostaglandin D2 receptor 1 antagonist, in microsomes and hepatocytes from preclinical species and humans. AB - (3R)-4-(4-Chlorobenzyl)-7-fluoro-5-(methylsulfonyl)-1,2,3,4 tetrahydrocyclopenta[b]indol-3-yl acetic acid (MK-0524) is a potent orally active human prostaglandin D(2) receptor 1 antagonist that is currently under development for the prevention of niacin-induced flushing. The major in vitro and in vivo metabolite of MK-0524 is the acyl glucuronic acid conjugate of the parent compound, M2. To compare metabolism of MK-0524 across preclinical species and humans, studies were undertaken to determine the in vitro kinetic parameters (K(m) and V(max)) for the glucuronidation of MK-0524 in Sprague-Dawley rat, beagle dog, cynomolgus monkey, and human liver microsomes, human intestinal microsomes, and in recombinant human UDP glucuronosyltransferases (UGT). A comparison of K(m) values indicated that UGT1A9 has the potential to catalyze the glucuronidation of MK-0524 in the liver, whereas UGT1A3 and UGT2B7 have the potential to catalyze the glucuronidation in the intestine. MK-0524 also was subject to phase I oxidative metabolism; however, the rate was significantly lower than that of glucuronidation. The rate of phase I metabolism was ranked as follows: rat approximately monkey > human intestine > dog > human liver with qualitatively similar metabolite profiles across species. In all the cases, the major metabolites were the monohydroxylated epimers (M1 and M4) and the keto metabolite, M3. Use of inhibitory monoclonal antibodies and recombinant human cytochromes P450 suggested that CYP3A4 was the major isozyme involved in the oxidative metabolism of MK-0524, with a minor contribution from CYP2C9. The major metabolite in hepatocyte preparations was the acyl glucuronide, M2, with minor amounts of M1, M3, M4, and their corresponding glucuronides. Overall, the in vivo metabolism of MK-0524 is expected to proceed via glucuronidation, with minor contributions from oxidative pathways. PMID- 17132766 TI - Insulin-like growth factor I in pregnancy and maternal risk of breast cancer. AB - BACKGROUND: The role of insulin-like growth factor (IGF)-I in breast cancer remains controversial, despite numerous reports on the association of the hormone with breast cancer or high-risk mammographic densities. We hypothesized that exposure to elevated IGF-I during early pregnancy, a period characterized by intense cell proliferation in the breasts and in the presence of high concentrations of sex steroids, will be associated with increased maternal risk to develop a breast malignancy. METHODS: The Northern Sweden Maternity Cohort is an ongoing prospective study, collecting blood samples from first-trimester pregnant women since 1975 as part of screening for infectious diseases. A case control study (212 cases and 369 controls) was nested among Northern Sweden Maternity Cohort members who delivered singleton babies. RIA was used to measure IGF-I and IGF-II levels. Conditional logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (95% CI). RESULTS: Breast cancer risk increased with increasing IGF-I (top tertile OR, 1.7; 95% CI, 1.1-2.7). The association was stronger among the primiparous (OR, 2.2; 95% CI, 1.1-4.4) than in the nonprimiparous women (OR, 1.4; 95% CI, 0.7-2.8). Upper-tertile risks seemed to decrease within the <28-, 28 to 33, and >33-year groups of age at sampling, from 2.5 (0.9-7.6) to 2.1 (0.9-5.0) and 1.2 (0.5-2.5), respectively. There was no association of breast cancer with first-trimester-pregnancy IGF-II. CONCLUSIONS: The study offers further evidence that IGF-I is important in breast cancer. Our findings suggest that the adverse effect of IGF-I on the breast may be stronger before the remodeling of the gland induced by a first pregnancy. PMID- 17132767 TI - Smoking relapse during the first year after treatment for early-stage non-small cell lung cancer. AB - BACKGROUND: Non-small-cell lung cancer patients who continue to smoke after cancer diagnosis are more likely to experience disease recurrence, decreased treatment efficacy, and treatment complications. Despite this, many continue to smoke, with estimates ranging from 13% to approximately 60%. METHODS: Participants were 154 early-stage, non-small-cell lung cancer patients who had smoked within 3 months before surgery. Patients were followed for 12 months after surgery to assess smoking status and duration of continuous abstinence after surgery. Predictors included medical, smoking history, psychosocial, and demographic characteristics. RESULTS: At some point after surgery, 42.9% of patients smoked; at 12 months after surgery, 36.9% were smoking. Sixty percent of patients who lapsed did so during the first 2 months after surgery. Smoking at follow-up was predicted by shorter quit duration before surgery, more intense Appetitive cravings (expectation of pleasure from smoking), lower income, and having a higher level of education. Time until the first smoking lapse was predicted by shorter quit duration before surgery, more intense Appetitive cravings to smoke, and lower income. Among those who lapsed, greater delay before the lapse was associated with abstinence at the 12-month follow-up assessment. CONCLUSIONS: Nearly half of non-small-cell lung cancer patients return to smoking after surgery if they have recent smoking histories. Most initial lapses happen within 2 months and occur in response to more recent smoking and more intense cravings. Findings suggest that interventions to prevent relapse should target those who wait until cancer surgery to quit smoking and should be started as soon as possible after treatment. PMID- 17132768 TI - Cancer incidence and mortality and vitamin D in black and white male health professionals. AB - Blacks have been documented to have low vitamin D levels. We thus examined whether total cancer incidence and mortality rates differ between Blacks and Whites in a population of male health professionals, and particularly for digestive system cancers (oral, esophagus, stomach, pancreas, and colorectum), which have been most consistently linked to poor vitamin D status. Second, we examined whether Blacks might be more susceptible to these cancers if they concurrently had other risk factors for hypovitaminosis D. In the Health Professionals Follow-up Study, from 1986 to 2002, 99 of 481 Black men and 7,019 of 43,468 White men were diagnosed with cancer. Adjusting for multiple dietary, lifestyle, and medical risk factors, using Cox modeling, Black men were at higher risk of total cancer incidence [relative risk (RR), 1.32; 95% confidence interval (95% CI), 1.08-1.61; P = 0.007] and total cancer mortality (RR, 1.89; 95% CI, 1.40-2.56; P < 0.0001) and especially digestive system cancer mortality (RR, 2.24; 95% CI, 1.35-3.70). Compared with Whites with relatively few risk factors for hypovitaminosis D, Blacks also with few risk factors for hypovitaminosis D were not at appreciably higher risk of total cancer incidence (RR, 0.95; 95% CI, 0.60-1.51) or mortality (RR, 1.55; 95% CI, 0.91-2.62), but Black men with additional risk factors for poorer vitamin D status had a much higher cancer incidence (RR, 1.57; 95% CI, 1.16-2.11) and mortality risk (RR, 2.27; 95% CI, 1.57-3.28). This pattern was even more pronounced for digestive system cancer. Our results suggest that the high frequency of hypovitaminosis D in Blacks may be an important, and easily modifiable, contributor to their higher risk of cancer incidence and mortality. PMID- 17132769 TI - Prolonged prevention of squamous cell carcinoma of the skin by regular sunscreen use. AB - Half of all cancers in the United States are skin cancers. We have previously shown in a 4.5-year randomized controlled trial in an Australian community that squamous cell carcinomas (SCC) but not basal cell carcinomas (BCC) can be prevented by regular sunscreen application to the head, neck, hands, and forearms. Since cessation of the trial, we have followed participants for a further 8 years to evaluate possible latency of preventive effect on BCCs and SCCs. After prolonged follow-up, BCC tumor rates tended to decrease but not significantly in people formerly randomized to daily sunscreen use compared with those not applying sunscreen daily. By contrast, corresponding SCC tumor rates were significantly decreased by almost 40% during the entire follow-up period (rate ratio, 0.62; 95% confidence interval, 0.38-0.99). Regular application of sunscreen has prolonged preventive effects on SCC but with no clear benefit in reducing BCC. PMID- 17132770 TI - Risk of multiple myeloma following medication use and medical conditions: a case control study in Connecticut women. AB - BACKGROUND: Certain commonly used drugs and medical conditions characterized by chronic immune dysfunction and/or antigen stimulation have been suggested to affect important pathways in multiple myeloma tumor cell growth and survival. We conducted a population-based case-control study to investigate the role of medical history in the etiology of multiple myeloma among Connecticut women. METHODS: A total of 179 incident multiple myeloma cases (21-84 years, diagnosed 1996-2002) and 691 population-based controls was included in this study. Information on medical conditions, medications, and medical radiation was obtained by in-person interviews. We calculated odds ratios (OR) as measures of relative risks using logistic regression models. RESULTS: A reduced multiple myeloma risk was found among women who had used antilipid statin therapy [OR, 0.4; 95% confidence interval (95% CI), 0.2-0.8] or estrogen replacement therapy (OR, 0.6; 95% CI, 0.4-0.99) or who had a medical history of allergy (OR, 0.4; 95% CI, 0.3-0.7), scarlet fever (OR, 0.5; 95% CI, 0.2-0.9), or bursitis (OR, 0.4; 95% CI, 0.2-0.7). An increased risk of multiple myeloma was found among women who used prednisone (OR, 5.1; 95% CI, 1.8-14.4), insulin (OR, 3.1; 95% CI, 1.1-9.0), or gout medication (OR, 6.7; 95% CI, 1.2-38.0). CONCLUSIONS: If our results are confirmed, mechanistic studies examining how prior use of insulin, prednisone, and, perhaps, gout medication might promote increased occurrence of multiple myeloma and how antilipid statins, estrogen replacement therapy, and certain medical conditions might protect against multiple myeloma may provide insights to the as yet unknown etiology of multiple myeloma. PMID- 17132771 TI - Functional studies of individual myosin molecules. AB - The "conventional" isoform of myosin that polymerizes into filaments (myosin II) is the molecular motor powering contraction in all three types of muscle. Considerable attention has been paid to the developmental progression, isoform distribution, and mutations that affect myocardial development, function, and adaptation. Optical trap (laser tweezer) experiments and various types of high resolution fluorescence microscopy, capable of interrogating individual protein motors, are revealing novel and detailed information about their functionally relevant nanometer motions and pico-Newton forces. Single-molecule laser tweezer studies of cardiac myosin isoforms and their mutants have helped to elucidate the pathogenesis of familial hypertrophic cardiomyopathies. Surprisingly, some disease mutations seem to enhance myosin function. More broadly, the myosin superfamily includes more than 20 nonfilamentous members with myriad cellular functions, including targeted organelle transport, endocytosis, chemotaxis, cytokinesis, modulation of sensory systems, and signal transduction. Widely varying genetic, developmental and functional disorders of the nervous, pigmentation, and immune systems have been described in accordance with these many roles. Compared to the collective nature of myosin II, some myosin family members operate with only a few partners or even alone. Individual myosin V and VI molecules can carry cellular vesicular cargoes much farther distances than their own size. Laser tweezer mechanics, single-molecule fluorescence polarization, and imaging with nanometer precision have elucidated the very different mechano-chemical properties of these isoforms. Critical contributions of nonsarcomeric myosins to myocardial development and adaptation are likely to be discovered in future studies, so these techniques and concepts may become important in cardiovascular research. PMID- 17132772 TI - Recruitment of new cells into the postnatal heart: potential modification of phenotype by periostin. AB - Establishment of the circulatory system occurs very early in development to support the rapid growth of the embryo. Therefore, the heart is the first functional organ to be formed during both avian and mammalian development. Historically, cardiac development has been considered to occur only during embryogenesis from cell sources located within the primordial structures that generate the myocardium and associated coronary vascular endothelium and smooth muscle and cardiac fibroblasts. Recently, however, contribution to the cardiac structures has been demonstrated to occur during embryonic development from extracardiac sources, like the anterior heart field, raising questions as to whether cardiogenesis may be an ongoing process that extends into adult life. In this brief article, we describe the contribution of circulating adult bone marrow hematopoietic stem cells to the cardiac cell populations and the potential regulation of their differentiation by the extracellular matrix protein, periostin. PMID- 17132773 TI - Cell-based approaches for cardiac repair. AB - Many forms of cardiovascular disease are associated with cardiomyocyte loss via necrosis and/or apoptosis. The cumulative loss of contractile cells ultimately results in diminished cardiac function. Numerous approaches have been employed to reduce the rate of cardiomyocyte loss, or alternatively, to repopulate the heart with new cardiomyocytes. Strategies aimed at repopulating the heart include cardiomyocyte cell therapy, myogenic stem cell therapy, and cell cycle activation therapy. All three approaches are based on the assumption that the de novo cardiomyocytes will participate in a functional syncytium with the surviving myocardium. This review will discuss the current status of interventions aimed at repopulating the heart with functional cardiomyocytes. PMID- 17132774 TI - The unstoppable connexin43 carboxyl-terminus: new roles in gap junction organization and wound healing. AB - Intercellular connectivity mediated by gap junctions (GJs) composed of connexin43 (Cx43) is critical to the function of excitable tissues such as the heart and brain. Disruptions to Cx43 GJ organization are thought to be a factor in cardiac arrhythmias and are also implicated in epilepsy. This article is based on a presentation to the 4th Larry and Horti Fairberg Workshop on Interactive and Integrative Cardiology and summarizes the work of Gourdie and his lab on Cx43 GJs in the heart. Background and perspective of recently published studies on the function of Cx43-interacting protein zonula occludens-(ZO)-1 in determining the organization of GJ plaques are provided. In addition how a peptide containing a PDZ-binding sequence of Cx43, developed as part of the work on cardiac GJ organization is also described, which has led to evidence for novel and unexpected roles for Cx43 in modulating healing following tissue injury. PMID- 17132775 TI - The developing cardiac myocyte: maturation of excitability and excitation contraction coupling. AB - The study of cardiac myocyte (CM) differentiation, development, and maturation is of interest for several compelling reasons. First, mechanisms of development are of fundamental biological interest. Second, congenital malformation of the heart may be related to CM dysfunction during embryonic/fetal development. Third, adult myocardium in a variety of diseased states re-expresses a fetal-like gene program. Fourth, the mature heart cannot readily regenerate itself. Thus, cell replacement therapy is an emerging treatment paradigm. Among the obstacles for the realization of cell replacement therapy is our incomplete understanding of the function during CM maturation. This is crucial in the potential use of embryonic stem (ES) cell-derived CMs as a cell source. Although much progress has been realized with mouse ES-CMs, our understanding of human counterparts is scant. Here we discuss key molecular underpinnings of excitability and excitation contraction coupling in developing mouse heart. We focus on the Ca channel multimeric complex and Ca handling. We compare mouse embryonic physiology to that previously described in mouse ES-CMs and draw parallels and highlight distinctions to human ES-CMs. During mouse embryonic and fetal maturation, the L type Ca channel current (I(Ca,L)) predominates, but embryonic/fetal I(Ca,L) has distinct properties from mature I(Ca,L). In addition T-type Ca current (I(Ca,T)) present in the fetus is not present in the adult. It is neither ethical nor practical to experiment with live human embryonic/fetal CMs for I(Ca) and Ca handling studies, but we can draw inferences from human heart cell function based on studies of human ES-CMs, using the parallels noted between mouse embryonic heart cells and mouse ES-CMs. PMID- 17132776 TI - Dynamic interactions between myocytes, fibroblasts, and extracellular matrix. AB - Cardiac function is determined by the coordinated and dynamic interaction of several cell types together with components of the extracellular matrix (ECM). This interaction is regulated by mechanical, chemical, and electrical signals between the cellular and noncellular components of the heart. Recent studies using fluorescence-activated cell sorting indicate that the number of myocytes remains relatively constant during development and disease, whereas the number of fibroblasts and other cell types can change dramatically. Cardiac fibroblasts appear to have different origins at different stages of development and fluctuate in response to a variety of physiological signals. Fibroblasts form a network of cells that are connected to each other via specific cadherins and connexins, to the ECM via integrins, and to myocytes by a variety of receptors, including connexins. Examples of the integration of signals include the role of angiotensin II (Ang II), which stimulates mechanical contraction of fibroblasts, as well as cytokine signaling. Cytokine signaling alters connexin and K(+) channel activation, which in turn is regulated by Ang II, essentially forming a feedback loop. Quantitative changes in mechanical, chemical, and electrical signals that can alter the overall cardiac form and function will be discussed here. PMID- 17132777 TI - Embryonic heart induction. AB - We have characterized two signaling pathways that induce heart tissue during embryonic development. The first is initiated by the Wnt antagonist Dickkopf1 (Dkk1) and involves the homeodomain transcription factor Hex. Other Wnt antagonists are less effective and the potency of Dkk1 might be due to synergy between Wnt antagonizing and another, novel activity emanating from its amino terminal cysteine-rich domain. The second signal is initiated by Nodal and its co receptor Cripto. Importantly, both the Dkk1/Wnt antagonism and Nodal pathways act on the endoderm that underlies the future heart to control secretion of diffusible factors that induce cardiogenesis in adjacent mesoderm. In this article, we summarize data that Dkk1 induces cardiogenic differentiation cell non autonomously through the action of the homeodomain transcription factor Hex. We also discuss recent data showing that Nodal also acts indirectly through stimulation of the secreted protein Cerberus, which is a member of the differential-screening selected aberrant in neuroblastoma (DAN) family of secreted proteins. Finally, we present the model that signaling from Dkk1 regulates novel activities, in addition to Wnt antagonism, which are essential for progression beyond initiation of cardiogenesis to control later stages of cardiomyocyte differentiation and myocardial tissue organization. PMID- 17132778 TI - The role of basic leucine zipper protein-mediated transcription in physiological and pathological myocardial hypertrophy. AB - Accumulating evidence suggests that nuclear transcription factors from the basic leucine zipper (bZIP) family play an important role in cardiac development and function. This class includes the CREB/ATF family of transcription factors, namely CREB, cAMP response element modulator (CREM), ATF, and the related AP-1 and C/EBP families. An effort has been made to elucidate the role of specific bZIP members in the heart. Unfortunately, little insight could be gained from knockout experiments, either due to embryonic lethal phenotypes or functional compensation by other bZIP family members. Surprisingly, cardiac overexpression of several inhibitory transcription factors from the bZIP family, such as a nonphosphorylatable form of CREB (CREB(ser133)), a nonfunctional isoform of CREM, or ATF3 resulted in massive atrial dilatation. In order to try and characterize this pathway we have expressed the potent bZIP inhibitory protein, Jun dimerization protein 2 (JDP2), specifically in the mouse heart in a temporally controlled manner. Expression of JDP2 resulted in massive biatrial dilatation; loss of connexin 40 (Cx40), connexin43 (Cx43), and myosin light chain 2 (MLC2a) expression; atrioventricular defects in conduction; and a lethal phenotype. All these effects were independent of any developmental events acquired during adulthood, and were totally reversible upon abolishing the bZIP inhibition. The results of this article suggest that bZIP inhibition is sufficient to cause atrial dilation, that this dilatation is acquired postnatally, and that it is reversible upon the relief of inhibition. Thus, bZIP repressors may serve as novel drug targets for the prevention of atrial dilatation a major risk of atrial fibrillation (AF). PMID- 17132779 TI - Hypertrophy and atrophy of the heart: the other side of remodeling. AB - The size of a cardiomyocyte is determined by relative rates of protein synthesis and degradation. Signaling pathways regulating myocardial protein synthesis have been extensively investigated, not the least because in patients hypertrophy increases cardiovascular morbidity and mortality. Until now strategies to reverse hypertrophy have relied on the inhibition of prohypertrophic signaling pathways. Here we review signaling pathways of atrophy in the heart and we present evidence in support of the idea that activating proatrophic signaling pathways in the presence of prohypertrophic signaling may be an attractive strategy to reverse hypertrophy. PMID- 17132780 TI - Role of cellular compartmentation in the metabolic response to stress: mechanistic insights from computational models. AB - The mechanisms controlling ATP generation in the transition from normal resting conditions to either high work states or ischemia are poorly understood. ATP generation depends upon compartmentation between the mitochondria and cytosol of metabolic pathways and key energy transfer species that cannot be easily assessed experimentally. We developed a multicompartment mathematical model of cardiac metabolism to simulate the metabolic responses to ischemia and increased workload. The model is based on mass balances, transport, and metabolic processes in cardiac tissue, and has three distinct compartments (blood, cytosol, and mitochondria). In addition to distinguishing between cytosol and mitochondria, the model includes a cytosolic subcompartment for glycolytic metabolic channeling. The model simulations predict the rapid activation of glycogenolysis and lactate production at the onset of ischemia, and support the concept of localization of glycolysis to a cytosolic subcompartment. In addition, simulations show that mitochondrial NADH/NAD(+) is primarily determined by oxygen consumption during ischemia, while cytosolic NADH/NAD(+) and lactate production are largely a function of glycolytic flux during the initial phase, and is controlled by mitochondrial NADH/NAD(+) and the malate-aspartate shuttle during the steady state. Finally, the model predicts that metabolic activation with an abrupt increase in workload requires parallel activation of ATP hydrolysis, glycolysis, mitochondrial dehydrogenases, the electron transport chain, and ADP phosphorylation. Taken together, these studies demonstrate the importance of metabolic compartmentation in the regulation of cardiac energetics in response to acute stress, and they highlight the usefulness of computational models in this line of investigation. PMID- 17132781 TI - Maintenance of the metabolic homeostasis of the heart: developing a systems analysis approach. AB - The heart is almost unique in the body with a constant requirement to conduct work well beyond the normal maintenance of cellular integrity. With this constant workload, it is not surprising that cardiac energy conversion is highly specialized to maintain a constant supply of energy. This maintenance of cellular metabolites during alterations in workload has been termed metabolic homeostasis. Here we discuss our efforts to understand the cellular and mitochondrial control network that orchestrates the metabolic homeostasis of the heart. This begins with a better definition of the metabolic pathways, acute posttranslational control sites, and proper kinetic evaluation of the reaction steps in the intact mitochondrial environment. First, a quantitative model of mitochondrial energy conversion is presented and demonstrates several serious gaps in our knowledge of this process. Toward filling these gaps, screens of the entire mitochondrial proteome have been conducted to establish the metabolic pathways that need to be considered. In addition, the dynamic phosphoproteome of intact mitochondria, using 2D gel electrophoresis coupled to (32)P labeling, has revealed a remarkably extensive protein phosphorylation network throughout the mitochondrial metabolic network that has essentially been overlooked. Initial studies on evaluating the functional significance of these protein phosphorylations and the kinase phosphatase system involved will be reviewed. One of the major deficits in the consensus quantitative model of oxidative phosphorylation to explain intact mitochondria activities is in complex I, where even the initiation of Nicotinamide Adenine Dinucleotide (reduced) (NADH) oxidation is problematical using in vitro kinetic data. Studies will be described where the NADH binding and oxidation kinetics at complex I in the intact mitochondria were determined using fluorescence lifetime and enzyme dependent-fluorescence recovery after photo oxidation (ED-FRAP) techniques. These later studies suggest that matrix NADH binding characteristics are much different (>10(3) binding constant errors) than isolated proteins. In addition, complex I is far from equilibrium and may play an important role in regulating the rate of reducing equivalent delivery to the cytochromes. PMID- 17132782 TI - Diversity of Ca2+ signaling in developing cardiac cells. AB - During embryonic and postnatal development, the mammalian heart undergoes rapid morphological changes with cellular differentiation that at the ultrastructural level encompasses altered expression and organization of the proteins and organelles associated with Ca(2+) signaling. Here the development and roles of the releasable Ca(2+) stores located within the sarco/endoplasmic reticulum and possibly within the nuclear envelopes are addressed. Confocal Ca(2+) imaging experiments were carried out on (i) neonatal rat cardiomyocytes, (ii) pluripotent P19 stem cells, differentiated to a cardiac phenotype by culturing with 1% dimethylsulfoxide (DMSO) in hanging droplets, and (iii) mouse embryonic cardiomyocytes isolated for short-time culture at embryonic day 9-18. The Ca(2+) release channels in neonatal and "cardiac" P19 cell were activated versus inhibited by targeting ryanodine (Ry) receptors with caffeine versus Ry and IP(3) receptors with adenosine 5'-triphosphate (ATP) or histamine versus U-73122, a phospholipase c (PLC) inhibitor. The neonatal cells displayed four recognizable phenotypes, of which two had specialized Ca(2+) stores releasable via either Ry or IP(3) receptors, and two had both types of receptors, either controlling functionally separate stores or with some degree of overlap, so that caffeine could deplete the stores releasable by ATP. The P19 cells showed variable presence of IP(3)-mediated Ca(2+) stores, and caffeine releasable stores that gained prominence in the "cardiac" phenotype, but were absent in a "neuronal" phenotype. The different roles of Ca(2+) stores were seen clearly in the mouse embryonic cells. Some cells from early stages of development (E 9-10) had Ca(2+) waves that increased in intensity during the diastolic interval and could trigger synchronous electrical excitation (via Na-Ca exchanger [NCX] and excitatory Ca(2+) and Na(+) channels). At later stages of development (E 18) we observed diastolic Ca(2+) sparks that appeared to originate from the nuclear envelope, while the Ca(2+) signals during excitation were faster and stronger in the nuclear region than in the surrounding cytoplasmic regions. However, we also found cells where the nuclear Ca(2+) signals were weaker and showed afterglow compared to the cytosolic Ca(2+) transients. We conclude that the Ca(2+) stores in cardiac cells during embryogenesis and postnatal development, that is, before the maturation of the t-tubular system and in stem cells with cardiac phenotype, show considerable diversity with respect to the pharmacology of the release channels and that regional differences in Ca(2+) signaling are observed centered in, at, and around the nucleus. We suggest that the causal relationship excitation and subcellular Ca(2+) signals in developing cardiac cells is different from that of adult cells and that the developing cardiomyocytes show a diversity that in later stages of development may be reflected in the different properties of atrial, ventricular, and pacemaker cells. PMID- 17132783 TI - Regulation of Ca2+ and Na+ in normal and failing cardiac myocytes. AB - Ca(2+) in cardiac myocytes regulates contractility and relaxation, and Ca(2+) and Na (+)regulation are linked via Na(+)/Ca(2+) exchange (NCX). Heart failure (HF) is accompanied by contractile dysfunction and arrhythmias, both of which may be due to altered cellular Ca(2+) handling. Smaller Ca(2+) transient and sarcoplasmic reticulum (SR) Ca(2+) content cause systolic dysfunction in HF. The reduced SR Ca(2+) content is due to: (a) reduced SR Ca(2+)-ATPase function (which also contributes to diastolic dysfunction), (b) increased expression and function of NCX (which competes with SR Ca(2+)-ATPase during relaxation, but preserves diastolic function), and (c) enhanced diastolic SR Ca(2+) leak. Relative contributions of these may vary with HF etiology and stage. Triggered arrhythmias (e.g., delayed afterdepolarizations [DADs]) are prominent in HF. DADs are due to spontaneous SR Ca(2+) release and consequent activation of transient inward NCX current, which in HF allows DADs to more readily trigger arrhythmogenic action potentials. Thus NCX and Na(+) are critical in systolic and diastolic function and arrhythmias. [Na(+)](i) is elevated in HF, which may limit SR unloading and provide some Ca(2+) influx during the HF action potential, thus limiting the depression of systolic function. High [Na(+)](i) in HF is due to enhanced Na(+) influx. Cellular Na(+)/K(+)-ATPase (NKA) function appears unaltered, despite reduced NKA expression. This dichotomy led us to test NKA regulation by phospholemman (PLM). We find that PLM regulates NKA in a manner analogous to phospholamban regulation of SR Ca(2+)-ATPase (i.e., inhibition that is relieved by PLM phosphorylation). We measured intermolecular FRET between PLM and NKA, which is reduced upon PLM phosphorylation. The lower expression level of more phosphorylated PLM in HF may explain the above dichotomy. Thus, altered Ca(2+) and Na(+) handling contributes to altered contractile function and arrhythmogenesis in HF. PMID- 17132784 TI - The integration of spontaneous intracellular Ca2+ cycling and surface membrane ion channel activation entrains normal automaticity in cells of the heart's pacemaker. AB - Although the ensemble of voltage- and time-dependent rhythms of surface membrane ion channels, the membrane "Clock", is the immediate cause of a sinoatrial nodal cell (SANC) action potential (AP), it does not necessarily follow that this ion channel ensemble is the formal cause of spontaneous, rhythmic APs. SANC also generates intracellular oscillatory spontaneous Ca(2+) releases that ignite excitation (SCaRIE) of the surface membrane via Na(+)/Ca(2+) exchanger activation. The idea that a rhythmic intracellular Ca(2+) Clock might keep time for normal automaticity of SANC, however, has not been assimilated into mainstream pacemaker dogma. Recent experimental evidence, derived from simultaneous, confocal imaging of submembrane Ca(2+) and membrane potential of SANC, and supported by numerical modeling, indicates that normal automaticity of SANC is entrained and stabilized by the tight integration of the SR Ca(2+) Clock that generates rhythmic SCaRIE, and the surface membrane Clock that responds to SCaRIE to immediately produce APs of an adequate shape. Thus, tightly controlled, rhythmic SCaRIE does not merely fine tune SANC AP firing, but is the formal cause of the basal and reserve rhythms, insuring pacemaker stability by rhythmically integrating multiple Ca(2+)-dependent functions, and effects normal automaticity by rhythmic ignition of the surface membrane Clock. PMID- 17132785 TI - Calcium handling in embryonic stem cell-derived cardiac myocytes: of mice and men. AB - Excitation-contraction (EC) coupling is fundamental to the function of cardiac myocytes (CMs). In mature myocytes plasma membrane (PM) L-type Ca(2+) channels function in close juxtaposition to ryanodine receptors (RyR) on the sarcoplasmic reticulum (SR) membrane. Action potentials (APs) cause the opening of PM L-type Ca(2+) channels, which in turn provide trigger Ca(2+) for a larger RyR-mediated SR Ca(2+) release. In contrast, developing myocytes have a less well-developed SR. This incomplete development is observed in early stage and mid-maturation stages of murine embryonic stem cell-derived cardiac myocytes (ESC-CMs). Despite the absence of a well-developed t-tubule system, murine ESC-CMs use internal Ca(2+) stores for EC coupling. Direct measures of Ca(2+) handling, including pharmacological studies and investigation of genetically modified mouse ESC-CMs, established an important contribution of RyR-mediated internal Ca(2+) store to cell function. Similarly, early-stage human ESC-CMs use internal Ca(2+) store and partially share Ca(2+) handling characteristics with murine ESC-CMs. For example, elementary Ca(2+) release events are present in both murine and human ESC-CMs, and it is likely that Ca(2+) handling contributes to automatic rhythm generation in these cells. However, in human ESC-CMs, a unique voltage-gated Na(+) channel window current is critical for spontaneous, rhythmic depolarization. The advent of the murine and human ES cardiomyocyte differentiating systems has provided initial insights into the early steps of development of excitability and electromechanical coupling in the mammalian heart, including patterns of gene expression, myofibrillogenesis, ion channel development and function, and Ca(2+) handling. Here we discuss the information gained from these models to describe the nexus of voltage-gated channel currents and Ca(2+) handling on rhythmic activity. PMID- 17132786 TI - Cellular alternans: a mechanism linking calcium cycling proteins to cardiac arrhythmogenesis. AB - Essentially all previous research on alternans has been restricted to normal myocardium, whereas sudden cardiac death (SCD) occurs most commonly in patients with ventricular dysfunction (i.e., heart failure), which is associated with marked disruption of proteins responsible for normal calcium cycling in myocytes. Several lines of evidence from studies in normal hearts suggest a link between impaired calcium cycling which characterizes ventricular mechanical dysfunction and impaired calcium cycling that is responsible for alternans. In normal myocardium, cells which exhibit the slowest calcium cycling, and not the slowest repolarization, are most susceptible to alternans. Decreased expression of key calcium cycling proteins is observed in alternans-prone cells. Sarcoplasmic reticulum ATPase (SERCA2a) expression is decreased, suggesting a mechanism for the slower sarcoplasmic reticulum (SR) calcium reuptake observed in alternans prone cells. In addition, diminished ryanodine receptor (RyR) function leading to abnormal calcium release from the SR is also linked to cellular alternans. Although impaired contractile function clearly predisposes to SCD, the mechanisms linking mechanical to electrophysiological dysfunction in the heart are unclear. We propose that cellular calcium alternans may be an important mechanism linking mechanical dysfunction to cardiac arrhythmogenesis. PMID- 17132787 TI - The mechanoelectric feedback: a novel "calcium clamp" method, using tetanic contraction, for testing the role of the intracellular free calcium. AB - Mechanical perturbations affect the membrane action potential, a phenomenon denoted as the mechanoelectric feedback (MEF), and may elicit cardiac arrhythmias. Two plausible mechanisms were suggested to explain this phenomenon: (i) stretch-activated channels (SACs) within the cell membrane and (ii) modulation of the action potential by the intracellular Ca(2+) (the Calcium hypothesis). The intracellular Ca(2+) varies mainly due to the effects of the mechanical perturbations on the affinity of troponin for calcium. The present study concentrates on the unique experimental methods that allow differentiating between the effects of SAC and Ca(2+) on the action potential. This is achieved by controlling the sarcomere lengths (SLs) independently of the intracellular Ca(2+) concentration, in the intact fiber. A dedicated experimental setup allowed simultaneous measurements of the membrane potential and the mechanical performance (Force and SL). The action potential was measured by voltage sensitive dye (Di-4-ANEPPS). The SL was measured by laser diffraction technique and was controlled by a fast servomotor. The intracellular Ca(2+) was controlled (calcium clamp) by imposing stable tetanic contractions at various extracellular calcium concentrations ([Ca(2+)](0)s). Tetanus was obtained by 8 Hz stimulation in the presence of cyclopiazonic acid (CPA) (30 muM). Isolated trabeculae from a rat's right ventricle were studied at different SLs and [Ca(2+)](0)s. The experimental data strongly support the calcium hypothesis. Although the action potential duration (APD) decreases at longer SL, the [Ca(2+)](0) has a significantly larger effect on the APD. The APD decreases as the [Ca(2+)](0) increases. Understanding the underlying mechanism opens new research avenues for the development of therapeutic modalities for cardiac arrhythmias. PMID- 17132788 TI - Role of sarcomere mechanics and Ca2+ overload in Ca2+ waves and arrhythmias in rat cardiac muscle. AB - Ca(2+) release from the sarcoplasmic reticulum (SR) depends on the sarcoplasmic reticulum (SR) Ca(2+) load and the cytosolic Ca(2+) level. Arrhythmogenic Ca(2+) waves underlying triggered propagated contractions arise from Ca(2+) overloaded regions near damaged areas in the cardiac muscle. Ca(2+) waves can also be induced in undamaged muscle, in regions with nonuniform excitation-contraction (EC) coupling by the cycle of stretch and release in the border zone between the damaged and intact regions. We hypothesize that rapid shortening of sarcomeres in the border zone during relaxation causes Ca(2+) release from troponin C (TnC) on thin filaments and initiates Ca(2+) waves. Elimination of this shortening will inhibit the initiation of Ca(2+) waves, while SR Ca(2+) overload will enhance the waves. Force, sarcomere length (SL), and [Ca(2+)](i) were measured and muscle length was controlled. A small jet of Hepes solution with an extracellular [Ca(2+)] 10 mM (HC), or HC containing BDM, was used to weaken a 300 mum long muscle segment. Trains of electrical stimuli were used to induce Ca(2+) waves. The effects of small exponential stretches on triggered propagatory contraction (TPC) amplitude and propagation velocity of Ca(2+) waves (V(prop)) were studied. Sarcomere shortening was uniform prior to activation. HC induced spontaneous diastolic sarcomere contractions in the jet region and attenuated twitch sarcomere shortening; HC+ butanedione monoxime (BDM) caused stretch only in the jet region. Stimulus trains induced Ca(2+) waves, which started inside the HC jet region during twitch relaxation. Ca(2+) waves started in the border zone of the BDM jet. The initial local [Ca(2+)](i) rise of the waves by HC was twice that by BDM. The waves propagated at a V(prop) of 2.0 +/- 0.2 mm/sec. Arrhythmias occurred frequently in trabeculae following exposure to the HC jet. Stretch early during relaxation, which reduced sarcomere shortening in the weakened regions, substantially decreased force of the TPC (F(TPC)) and delayed Ca(2+) waves, and reduced V(prop) commensurate with the reduction F(TPC). These results are consistent with the hypothesis that Ca(2+) release from the myofilaments initiates arrhythmogenic propagating Ca(2+) release. Prevention of sarcomere shortening, by itself, did not inhibit Ca(2+) wave generation. SR Ca(2+) overload potentiated initiation and propagation of Ca(2+) waves. PMID- 17132789 TI - Cellular basis for the repolarization waves of the ECG. AB - One hundred years after Willem Einthoven first recorded the electrocardiogram (ECG), physicians and scientists are still debating the cellular basis for the various waves of the ECG. In this review, our focus is on the cellular basis for the J, T, and U waves of the ECG. The J wave and T wave are thought to arise as a consequence of voltage gradients that develop as a result of the electrical heterogeneities that exist within the ventricular myocardium. The presence of a prominent action potential notch in epicardium but not endocardium gives rise to a voltage gradient during ventricular activation that inscribes the J wave. Transmural and apico-basal voltage gradients developing as a result of difference in the time course of repolarization of the epicardial, M, and endocardial cell action potentials, and the more positive plateau potential of the M cell contribute to inscription of the T wave. Amplification of these heterogeneities results in abnormalities of the J wave and T wave, leading to the development of the Brugada, long QT, and short QT syndromes. The basis for the U wave has long been a matter of debate. One theory attributes the U wave to mechanoelectrical feedback. A second theory ascribes it to voltage gradients within ventricular myocardium and a third to voltage gradients between the ventricular myocardium and the His-Purkinje system. Although direct evidence in support of any of these three hypotheses is lacking, recent studies involving the short QT syndrome have generated renewed interest in the mechanoelectrical hypothesis. PMID- 17132790 TI - Mechanosensitive-mediated interaction, integration, and cardiac control. AB - This review covers aspects of the cardiac mechanotransduction field at different levels, and advocates the possibility that mechanoelectro-chemical transduction forms part of a network of mechanically linked integration in heart-mechanically mediated integration (MMI). It assembles evidence and observations in the literature to promote this hypothesis. Mechanical components can provide the bond between interactions at molecular, cellular, and macro levels to enable the integration. Stretch-activated channels (SACs) exist in the heart, but stresses and strains can affect other membrane channels or receptors. A cellular mechanical change can thus promote several ionic or downstream changes. Cell signal cascades have been implicated and can affect membrane electrophysiology. MMI could shape intracellular and downstream signals using the cytoskeleton and intracellular Ca(2+). MMI also spans other regulatory systems and processes such as the autonomic nervous system (ANS) and operates throughout the whole heart as an integrative system. Finally, supporting the hypothesis, if elements of the normal integration become deranged it contributes to cardiovascular disease and, potentially, lethal arrhythmia. PMID- 17132791 TI - Three-dimensional models of individual cardiac histoanatomy: tools and challenges. AB - There is a need for, and utility in, the acquisition of data sets of cardiac histoanatomy, with the vision of reconstructing individual hearts on the basis of noninvasive imaging, such as MRI, enriched by reference to detailed atlases of serial histology obtained from representative samples. These data sets would be useful not only as a repository of knowledge regarding the specifics of cardiac histoanatomy, but could form the basis for generation of individualized high resolution cardiac structure-function models. The current article presents a step in this general direction: it illustrates how whole-heart noninvasive imaging can be combined with whole-heart histology in an approach to achieve automated construction of histoanatomically detailed models of cardiac 3D structure and function at hitherto unprecedented resolution and accuracy (based on 26.4 x 26.4 x 24.4 microm MRI voxel size, and enriched by histological detail). It provides an overview of the tools used in this quest and outlines challenges posed by the approach in the light of applications that may benefit from the availability of such data and tools. PMID- 17132792 TI - Cardiac defibrillation and the role of mechanoelectric feedback in postshock arrhythmogenesis. AB - Ventricular dilatation increases the defibrillation threshold (DFT). In order to elucidate the mechanisms responsible for this increase, the present article investigates changes in the postshock behavior of the myocardium upon stretch. A two-dimensional electro-mechanical model of cardiac tissue incorporating heterogeneous fiber orientation was used to explore the effect of sustained stretch on postshock behavior via (a) recruitment of mechanosensitive channels (MSC) and (b) tissue deformation and concomitant changes in tissue conductivities. Recruitment of MSC had no influence on vulnerability to electric shocks as compared to control, but increased the complexity of postshock VF patterns. Stretch-induced deformation and changes in tissue conductivities resulted in a decrease in vulnerability to electric shocks. PMID- 17132793 TI - Modeling cardiac electrical activity at the cell and tissue levels. AB - Significant tissue structures exist in cardiac ventricular tissue, which are of supracellular dimension. It is hypothesized that these tissue structures contribute to the discontinuous spread of electrical activation, may contribute to arrhythmogenesis, and also provide a substrate for effective cardioversion. However, the influences of these mesoscale tissue structures in intact ventricular tissue are difficult to understand solely on the basis of experimental measurement. Current measurement technology is able to record at both the macroscale tissue level and the microscale cellular or subcellular level, but to date it has not been possible to obtain large volume, direct measurements at the mesoscales. To bridge this scale gap in experimental measurements, we use tissue-specific structure and mathematical modeling. Our models, which can incorporate ion channel models at the cell level into the reaction-diffusion equations at the tissue level, have enabled us to consider key hypotheses regarding discontinuous activation. PMID- 17132794 TI - Cardiac beta-adrenergic signaling: from subcellular microdomains to heart failure. AB - beta-adrenergic signaling plays a central role in the neurohumoral regulation of the heart and the progression of heart failure. Initially thought to be a simple linear cascade, this complex network is now recognized to utilize cross-talk with numerous other pathways, spatial compartmentation, and feedback control to coordinate cardiac electrophysiology, contractility, and adaptive remodeling. Here, we review recent basic insights and novel quantitative approaches that are leading to a more comprehensive understanding of beta-adrenergic signaling and thus motivate new therapeutic strategies for cardiac disease. PMID- 17132795 TI - Multiscale modeling of calcium signaling in the cardiac dyad. AB - Calcium (Ca(2+))-induced Ca(2+)-release (CICR) takes place in spatially restricted microdomains known as dyads. The length scale over which CICR occurs is on the order of nanometers and relevant time scales range from micro- to milliseconds. Quantitative understanding of CICR therefore requires development of models that are applicable over a range of spatio-temporal scales. We will present several new approaches for multiscale modeling of CICR. First, we present a model of dyad Ca(2+) dynamics in which the Fokker-Planck equation (FPE) is solved for the probability P(x, t) that a Ca(2+) ion is located at dyad position x at time t. Using this model, we demonstrate that (a) Ca(2+) signaling in the dyad is mediated by approximately tens of Ca(2+) ions; (b) these signaling events are noisy due to the small number of ions involved; and (c) the geometry of the RyR (ryanodine receptors) protein may function to restrict the diffusion of and to "funnel" Ca(2+) ions to activation-binding sites on the RyR, thus increasing RyR open probability and excitation-contraction (EC) coupling gain. Simplification of this model to one in which the dyadic space is represented using a single compartment yields the stochastic local-control model of CICR developed previously. We have shown that this model captures fundamental properties of CICR, such as graded release and voltage-dependent gain, may be integrated within a model of the myocyte and may be simulated in reasonable times using a combination of efficient numerical methods and parallel computing, but remains too complex for general use in cell simulations. To address this problem, we show how separation of time scales may be used to formulate a model in which nearby L-type Ca(2+) channels (LCCs) and RyRs gate as a coupled system that may be described using low-dimensional systems of ordinary differential equations, thus reducing computational complexity while capturing fundamentally important properties of CICR. The simplified model may be solved many orders of magnitude faster than can either of the more detailed models, thus enabling incorporation into tissue-level simulations. PMID- 17132796 TI - Nonlinear dynamics of paced cardiac cells. AB - When a cardiac cell is rapidly paced it can exhibit a beat-to-beat alternation in the action potential duration (APD) and the intracellular calcium transient. This dynamical instability at the cellular level has been shown to correlate with the genesis of cardiac arrhythmias and has motivated the application of nonlinear dynamics in cardiology. In this article, we review mathematical approaches to describe the underlying mechanisms for alternans using beat-to-beat iterated maps. We explain the development and properties of these maps, and show that they provide a fruitful framework to understand dynamical instabilities of voltage and calcium in paced cardiac cells. PMID- 17132797 TI - Modeling cardiac ischemia. AB - Myocardial ischemia is one of the main causes of sudden cardiac death, with 80% of victims suffering from coronary heart disease. In acute myocardial ischemia, the obstruction of coronary flow leads to the interruption of oxygen flow, glucose, and washout in the affected tissue. Cellular metabolism is impaired and severe electrophysiological changes in ionic currents and concentrations ensue, which favor the development of lethal cardiac arrhythmias such as ventricular fibrillation. Due to the burden imposed by ischemia in our societies, a large body of research has attempted to unravel the mechanisms of initiation, sustenance, and termination of cardiac arrhythmias in acute ischemia, but the rapidity and complexity of ischemia-induced changes as well as the limitations in current experimental techniques have hampered evaluation of ischemia-induced alterations in cardiac electrical activity and understanding of the underlying mechanisms. Over the last decade, computer simulations have demonstrated the ability to provide insight, with high spatiotemporal resolution, into ischemic abnormalities in cardiac electrophysiological behavior from the ionic channel to the whole organ. This article aims to review and summarize the results of these studies and to emphasize the role of computer simulations in improving the understanding of ischemia-related arrhythmias and how to efficiently terminate them. PMID- 17132798 TI - Cardiovascular therapeutic aspects of cell therapy and stem cells. AB - The recent advancements in stem cell biology, molecular and cell biology, and tissue engineering have paved the way to the development of a new biomedical discipline: regenerative medicine. The heart represents an attractive candidate for this emerging discipline since these emerging technologies could be used to potentially treat a variety of myocardial disorders. Here we describe our efforts in using stem cell and cell therapy strategies to restore the myocardial electromechanical properties. Specifically, our research has focused on the potential role of human embryonic stem cells (hESC) for myocardial regeneration (for the treatment of heart failure) and on using genetically engineered cell grafts to modify the myocardial electrophysiological properties (for the treatment of cardiac arrhythmias). The recently described hESC lines are unique pluripotent cell lines that can be propagated in the undifferentiated state in culture and coaxed to differentiate into cell derivatives of all three germ layers, including cardiomyocytes. The current article describes this unique cardiomyocyte differentiating system and details the molecular, ultrastructural, and functional properties of the generated hESC-derived cardiomyocytes (hESC CMs). The ability of the hESC-CMs to integrate structurally and functionally with host cardiomyocytes in both in vitro and in vivo studies will be described as well as their ability to restore the myocardial electromechanical function in animal models of diseased hearts. We will next present detailed in vitro, in vivo, and computer simulation studies performed in our laboratory testing the hypothesis that cell grafts, engineered to express specific ion channels, can be used to modify the myocardial electrophysiological properties of cardiac tissue. The potential and drawbacks of this novel approach for the treatment of both tachyarrhythmias (using cell grafts expressing potassium channels) and bradyarrhythmias (using hESC coaxed to differentiate into pacemaking cells or conducting tissue) will be described. PMID- 17132799 TI - Adenylyl cyclase gene transfer in heart failure. AB - The rationale for gene transfer of adenylyl cyclase type VI (AC(VI)) for clinical congestive heart failure (CHF) is based on recent experimental studies that have extended from cultured cardiac myocytes to preclinical studies in animal models of CHF. Over the past several years substantial data have indicated an unexpected and pronounced favorable effect of AC(VI) expression in cardiovascular disease. Preclinical studies have shown that increased cardiac AC content improves left ventricular function and attenuates deleterious remodeling in the failing heart, and reduces mortality in heart failure and in acute myocardial infarction. A brief review of the preclinical studies that have examined changes associated with increased AC expression in the heart is presented here. PMID- 17132800 TI - Genetic engineering and therapy for inherited and acquired cardiomyopathies. AB - The cardiac myofilaments consist of a highly ordered assembly of proteins that collectively generate force in a calcium-dependent manner. Defects in myofilament function and its regulation have been implicated in various forms of acquired and inherited human heart disease. For example, during cardiac ischemia, cardiac myocyte contractile performance is dramatically downregulated due in part to a reduced sensitivity of the myofilaments to calcium under acidic pH conditions. Over the last several years, the thin filament regulatory protein, troponin I, has been identified as an important mediator of this response. Mutations in troponin I and other sarcomere genes are also linked to several distinct inherited cardiomyopathic phenotypes, including hypertrophic, dilated, and restrictive cardiomyopathies. With the cardiac sarcomere emerging as a central player for such a diverse array of human heart diseases, genetic-based strategies that target the myofilament will likely have broad therapeutic potential. The development of safe vector systems for efficient gene delivery will be a critical hurdle to overcome before these types of therapies can be successfully applied. Nonetheless, studies focusing on the principles of acute genetic engineering of the sarcomere hold value as they lay the essential foundation on which to build potential gene-based therapies for heart disease. PMID- 17132801 TI - Nanomedicine opportunities in cardiology. AB - Despite myriad advances, cardiovascular-related diseases continue to remain our greatest health problem. In more than half of patients with atherosclerotic disease, their first presentation to medical attention becomes their last. Patients often survive their first cardiac event through acute revascularization and placement of drug-eluting stents (DES), but only select coronary lesions are amenable to DES placement, resulting in the use of bare metal or no stent, both of which lack the benefit of antirestenotic therapy. In other patients, transient ischemic attacks (TIAs) and stroke constitute the initial presentation of disease. In these patients, the diagnostic and therapeutic options are woefully inadequate. Nanomedicine offers options to each of these challenges. Antiangiogenic paramagnetic nanoparticles may be used to serially assess the severity of atherosclerotic disease in asymptomatic, high-risk patients by detecting the development of plaque neovasculature, which reflects the underlying lesion activity and vulnerability to rupture. The nanoparticles can locally deliver antiangiogenic therapy, which may acutely retard plaque progression, allowing aggressive statin therapy to become effective. Moreover, these agents may be useful as a quantitative marker to guide atherosclerotic management in an asymptomatic patient. In those cases proceeding to the catheterization laboratory for revascularization, nanoparticles incorporating antirestenotic drugs can be delivered directly into the wall of lesions not amenable to DES placement. Targeted nanoparticles could help ensure that antirestenotic drugs are available for all lesions. Moreover, displacement of antiproliferative agents from the intimal surface into the vascular wall is likely to improve rehealing of the endothelium, improving postprocedural management of these patients. PMID- 17132802 TI - Effects of synchronized cardiac assist device on cardiac energetics. AB - A novel physiological cardiac assist device (PCAD), the LEV RAM assist device, which is synchronized with the failing heart ejection, was developed to improve the failing heart systolic and diastolic functions and cardiac energetics. The PCAD uses a single short cannula, which is inserted into the beating left ventricle (LV) by means of a specially designed device. Blood is ejected from the PCAD into the LV after the opening of the aortic valve and augments the cardiac stroke work. The same amount of blood is withdrawn from the LV into the PCAD, through the same cannula, during the diastole. The study aims to test the effects of the PCAD on cardiac energetics and coronary blood flow. Adult normal sheep were anesthetized and the heart was exposed by left thoracotomy. Pressures transducers (Millar Instruments, Inc., Houston, TX) were inserted into the LV and aorta. LV volume was measured by sonocrystals (Sonometrics Corp., London, Ontario, Canada) and impedance catheter (CD Lycom, Argonstrat 116 Zoetermeer, 2718 SP The Netherlands). Flowmeters (transonic) measured the cardiac output (CO) and the coronary arteries (left anterior descending (LAD) and circumflex) flows. A thin cannula was inserted into the coronary sinus and the oxygen content of the LV and the coronary sinus were determined (AVOXimeter-1000). Pressure-volume loops, myocardial energetics, and coronary flow were measured. The displaced PCAD volume was 11 mL. Four different levels of assist were studied by changing the frequency of the assist: (1) assist beat after three successive regular beats [1:4], (2) assist every third beat [1:3], (3) alternate assist and normal beat [1:2], and (4) continuous assist [1:1]. Cardiac output (CO) and stroke volume (SV) increased proportionally with increasing frequency of assist. Systolic mechanical efficiency of the PCAD was above 90%. Simultaneously, the PCAD decreased the end-diastolic volume (EDV; diastolic unloading). The PCAD increased coronary flow and decreased cardiac arterial-venous O(2) difference. We conclude that the PCAD efficiently augments CO and stroke work, decreases preload, and decreases the coronary arterial-venous O(2) difference; all these may expedite cardiac reverse remodeling, and promote recovery of function and eventual easy explanation of the device. PMID- 17132803 TI - Statins and progression of calcified aortic stenosis. AB - OBJECTIVE: To review the evidence evaluating the efficacy of statins in reducing the progression of calcified aortic stenosis (AS). DATA SOURCES: MEDLINE, EMBASE, and PubMed were searched (all up to November 2006) for studies evaluating the use of statins to reduce the progression of calcified AS. Search terms included statin, HMG CoA reductase inhibitor, calcified AS, valve stenosis, and calcified stenosis. Additional primary trials were located by searching references noted in review articles. STUDY SELECTION AND DATA EXTRACTION: Clinical trials published in the English language were selected for review. Primary efficacy outcomes evaluated were changes in aortic valve measurements, hemodynamic measures of AS, and change in measures of AS severity. DATA SYNTHESIS: Two prospective clinical trials and 5 retrospective studies were included in this review. All of the retrospective studies demonstrated that statin use was associated with a statistically significant delay in the progression of AS. One prospective observation trial showed benefit of statin use; however, a large, randomized, double-blind, prospective trial showed no benefit of statin use in decreasing the progression of AS. CONCLUSIONS: An association between statin use and a delay in AS progression has been observed in retrospective studies; however, prospective trials showed conflicting results. Currently, statins cannot be recommended for medical treatment of AS until larger trials are conducted. PMID- 17132804 TI - Dosage adjustments according to renal function at discharge: a comparison of 3 hospitals. PMID- 17132805 TI - Improving oral medicine administration in patients with swallowing problems and feeding tubes. AB - BACKGROUND: Correct administration of oral medicines in patients with swallowing problems and feeding tubes remains a challenge. OBJECTIVE: To improve drug administration in patients with swallowing problems and feeding tubes in a 1600 bed tertiary referral center in the State of Qatar. METHODS: A questionnaire was used to evaluate the knowledge and practice of nursing staff from 6 different intensive care units (ICU). Questions assessed the respondents' knowledge of the purpose of controlled-release (CR) preparations, codes used for CR medication, the consequences of crushing these preparations, and their interactions with enteral feeds and feeding tubes. Based on the results of the questionnaire, a 2 day training program was conducted for a core group of 34 staff nurses from all units (ICU and non-ICU) and 3 nursing instructors. Following the principle of "training the trainers," the core group and the nursing instructors would widen the scope of education by running programs in the future. Lecture sessions for pharmacy staff and a monitoring tool to evaluate the practice were also developed. RESULTS: Overall knowledge of CR codes increased from 0% to 40%, correct crushing of solid preparations from 35% to 90%, and knowledge of possible interactions with the enteral feed or feeding tubes from 51% to 88%. Correct administration of medication in patients with feeding tubes improved from 32% to 83%. The quality and value of the 2 day training course received a mean score of 96%. CONCLUSIONS: While most nurses were aware of the purpose of CR formulations, little was known about the different codes used by drug companies and the consequences of crushing these preparations. Interactions with enteral feeds and feeding tubes were mostly overlooked. A 2 day training course followed by continuous in-service training, a lecture to pharmacy staff, and provision of supportive educational material greatly improved the overall medicine administration process in patients with swallowing problems and feeding tubes. PMID- 17132806 TI - Clozapine effective in olanzapine-induced Pisa syndrome. AB - OBJECTIVE: To report a case of olanzapine-induced Pisa syndrome that improved after treatment with clozapine. CASE SUMMARY: A 22-year-old male with paranoid schizophrenia presented with insidious onset tonic truncal flexion with axial rotation and difficulty in walking after exposure to olanzapine in doses up to 15 mg/day for 9 months. An objective causality assessment suggested that Pisa syndrome was probably related to olanzapine. There was improvement in his symptoms after 6 weeks of treatment with clozapine in doses gradually titrated to 350 mg/day. DISCUSSION: Pisa syndrome is a type of dystonia that has been associated with both typical and atypical antipsychotics. Both acute and insidious onset cases have been described in the literature, which have different course and treatment response. Clozapine was found to be effective in reducing the severity of olanzapine-induced Pisa syndrome. CONCLUSION: Clozapine may be a useful treatment option for Pisa syndrome that has been caused by olanzapine. PMID- 17132807 TI - Androgen deprivation in veterans with prostate cancer: implications for skeletal health. AB - BACKGROUND: Common risk factors for osteoporosis in older men include smoking, heavy use of alcohol, propensity to falls, and use of bone-toxic medications such as prednisone. There is also increasing appreciation of the skeletal risk faced by men receiving androgen deprivation therapy (ADT) for prostate cancer. Measures to prevent bone loss in such patients are available. OBJECTIVE: To test the following hypotheses in a population of veterans receiving ADT for prostate cancer: (1) fracture risk factors in addition to androgen deprivation would be found in most patients, (2) bone mass measurements would be assessed in a minority of patients, and (3) a minority of the subjects would receive bisphosphonate therapy or have contraindications for such treatment. METHODS: We conducted a retrospective chart review of male veterans receiving ADT from 1993 through 2001, at the Veterans Affairs Medical Center, Madison, WI. RESULTS: One hundred and seventy-four subjects met study criteria, with a mean age of 76 years and median duration of 21 months of ADT. Eighty-one percent had risk factors in addition to ADT. Only 13% underwent bone density measurement by dual energy X-ray absorptiometry (DXA) and, of those measured, more than half had osteoporosis. Only 19% of the men received both calcium and vitamin D supplements. Antiresorptive therapy was provided to 11% of men, although more than two-thirds had no contraindications to therapy. A total of 24 men sustained a fracture after starting ADT. For men who did undergo bone density measurement, 77% received antiresorptive therapy. Of those who exhibited osteoporosis by DXA scan, 85% received antiresorptive therapy. CONCLUSIONS: Male veterans receiving ADT for prostate cancer received inadequate evaluation and treatment for osteoporosis. Based on our data, a simple and practical strategy to prompt further evaluation and improved care may be to undertake bone density measurements in men prior to or soon after commencing ADT. PMID- 17132808 TI - Extracorporeal removal of vancomycin by plasmapheresis. PMID- 17132809 TI - Breast-feeding during maternal use of azathioprine. AB - OBJECTIVE: To report the clinical outcome of infants whose mothers were taking azathioprine while nursing and to quantify the transfer of 6-mercaptopurine (6 MP), its active metabolite, into breast milk. CASE SUMMARY: We report on a series of 4 patients treated with azathioprine while lactating. Breast milk samples were analyzed for 6-MP in 2 of the mothers. Several milk samples per patient were analyzed; levels of 6-MP were undetectable by high performance liquid chromatography (limit of detection 5 ng/mL). Therefore, the absolute relative infant dose would have been less than 0.09% of the maternal weight-adjusted dose. No adverse effects were encountered in any of the 4 infants. DISCUSSION: A large number of women of reproductive age are treated with azathioprine for a range of chronic conditions that require immunosuppression, such as systemic lupus erythematosus or solid organ transplants. Similar to other antimetabolites, the drug has generally been contraindicated for use during breast-feeding because of the theoretical concern for toxicity in the nursing infant. The available literature in this area is sparse and dated. The data presented here confirm published reports of minimal 6-MP excretion into milk, suggesting that significant systemic adverse effects in the infant are unlikely. CONCLUSIONS: Maternal azathioprine use during lactation does not appear to pose a significant immediate clinical risk to the suckling infant. Continued monitoring and long term assessment of these infants are warranted. PMID- 17132810 TI - Febuxostat: a selective xanthine oxidase inhibitor for the treatment of hyperuricemia and gout. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, clinical trial data, safety profile, precautions, and place in therapy of febuxostat, a novel nonpurine xanthine oxidase inhibitor in development for the treatment of hyperuricemia and gout. DATA SOURCES: Available studies and abstracts were identified through MEDLINE (1990-November 2006), Science Citation Index, International Pharmaceutical Abstracts, Cochrane Databases, and the American College of Rheumatology and European League Against Rheumatism Web sites. Key search terms were febuxostat, TMX-67, TEI-6720, hyperuricemia, and gout. STUDY SELECTION AND DATA EXTRACTION: All available studies describing the pharmacology of febuxostat were included. Human studies formed the basis for discussion of clinical parameters, including pharmacokinetics, pharmacodynamics, efficacy, and safety of febuxostat. DATA SYNTHESIS: Febuxostat significantly reduces uric acid levels within 2 weeks after initiation of therapy and up to 48% by the end of 104 weeks of therapy. Approximately 60% of patients achieved the primary goal of serum uric acid less than 6 mg/dL during the last 3 months following once-daily administration of febuxostat 80 mg or 120 mg for at least 52 weeks. The most common adverse reactions to febuxostat were abnormal results from liver function tests, diarrhea, headache, arthralgias, and musculoskeletal symptoms. Due to its potency, patients are at an increased risk of experiencing gout flares for at least the first year of therapy. Up to 70% of patients in clinical trials experienced gout flares despite concomitant prophylactic treatment with colchicine or naproxen. Additional clinical trial evidence supports the efficacy and safety of febuxostat in the treatment of hyperuricemia and gout. CONCLUSIONS: Febuxostat is a promising alternative to allopurinol for the treatment of gout and hyperuricemia. The optimal length of prophylactic therapy, clinical significance of abnormal liver function tests results during therapy, and safety in patients with moderate or severe renal insufficiency warrant further investigation. PMID- 17132811 TI - Relationship between pharmaceutical company user fees and drug approvals in Canada and Australia: a hypothesis-generating study. AB - BACKGROUND: Since the early- to mid-1990s, drug companies have paid fees for a variety of activities carried out by the Therapeutic Products Directorate in Canada and the Therapeutic Goods Administration in Australia. OBJECTIVE: To explore whether changes in approval times for new active substances and in the percentage of new drug submissions receiving positive decisions coincided with the level of user fees. METHODS: Data were collected from a range of Canadian and Australian government publications on the following topics: total funding for and workload of the regulatory agencies, the percentage of income that came from tax revenue and user fees, the percentage of new drug submissions that received a positive decision, and-for Canada only-the percent of submissions that were approved on first review. RESULTS: In both countries, there was a moderate-to strong positive association between the level of industry funding and the percent of submissions that received a positive decision and a moderate-to-strong (Canada) and moderate (Australia) negative association between the level of industry funding and approval times. CONCLUSIONS: Changes observed in both countries are favorable to the pharmaceutical industry. Other than user fees leading to a pro-industry bias in the regulatory authorities, other possible explanations include a more efficient use of resources, a smaller workload (Canada), an improvement in the quality of drug submissions (Canada), and more resources (Australia). Further research strategies are needed to either confirm or refute the hypothesis that the level of industry funding affects decisions made in drug regulatory systems. PMID- 17132812 TI - Comment: paroxetine use during pregnancy: is it safe? PMID- 17132813 TI - Role of adenosine receptors in the regulation of angiogenic factors and neovascularization in hypoxia. AB - Because hypoxia increases extracellular adenosine levels and stimulates angiogenesis, we evaluated the relative roles of reduced oxygen concentrations and adenosine receptor activation in the production of angiogenic factors. In vitro, we analyzed the effects of hypoxia and adenosine on the secretion of angiogenic factors from human microvascular endothelial cells (HMEC-1). To study the effects of hypoxia alone, we scavenged adenosine from the hypoxic medium with adenosine deaminase, and we used the stable adenosine analog 5'-N ethylcarboxamidoadenosine (NECA) to study the effects of stimulation of adenosine receptors. In the absence of adenosine, hypoxia stimulated vascular endothelial growth factor (VEGF) but not interleukin-8 (IL-8) secretion from HMEC-1. In contrast, NECA stimulated both VEGF and IL-8 secretion. VEGF secretion was increased 1.9 +/- 0.04-fold with NECA (10 microM) and 1.7 +/- 0.1-fold with hypoxia (5% O(2)) but 3.8 +/- 0.1-fold when these two stimuli were combined. Thus, adenosine receptors act in a cooperative fashion with hypoxia to stimulate VEGF and induce IL-8 secretion not stimulated by hypoxia alone. In vivo, antagonism of adenosine receptors with caffeine abrogated VEGF up-regulation induced by local injection of NECA into the mouse hind limb and produced a 46% reduction of neovascularization in a mouse ischemic hind limb model. Our study suggests that adenosine actions are not redundant but rather are complementary to the direct effects of hypoxia. Stimulation of adenosine receptors not only contributes to the overall effect of hypoxia but also has additional actions in the regulation of angiogenic factors. Thus, adenosine receptors represent a potential therapeutic target for regulation of neovascularization. PMID- 17132814 TI - Secoisolariciresinol diglucoside: relevance to angiogenesis and cardioprotection against ischemia-reperfusion injury. AB - Therapeutic angiogenesis represents a novel approach for the prevention and treatment of ischemic heart disease. This study examined a novel method of stimulating myocardial angiogenesis using secoisolariciresinol diglucoside (SDG), a plant lignan isolated from flaxseed. SDG has been shown to decrease serum cholesterol and reduce the extent of atherosclerosis. In the present study, the angiogenic properties of SDG were investigated in three different models. First, in the in vitro model, human coronary arteriolar endothelial cells (HCAEC) treated with SDG (50 and 100 microM) showed a significant increase in tubular morphogenesis compared with control. Western blot analysis indicated an increased expression of vascular endothelial growth factor (VEGF), kinase insert domain containing receptor (KDR), Flt-1, angiopoietin-1 (Ang-1), Tie-1, and phosphorylated endothelial nitric oxide synthase (p-eNOS) in the SDG-treated cells. Second, in the ex vivo ischemia/reperfusion model, SDG-treated rats (20 mg/kg b.wt./day for 2 weeks orally) showed an increased level of aortic flow and functional recovery after 2 h of reperfusion following 30 min of ischemia compared with the control group [dP/dt (mm Hg/s) of 2110 +/- 35 versus 1752 +/- 62]. SDG reduced infarct size compared with the control group by 32% (38 versus 26%) and also decreased cardiomyocyte apoptosis. Increased protein expression of VEGF, Ang-1, and p-eNOS was also observed in the SDG-treated group. Third, in the in vivo myocardial infarction model, SDG increased capillary density and myocardial function as evidenced by increased fractional shortening and ejection fraction. In conclusion, these results suggest that SDG has potent angiogenic and antiapoptotic properties that may contribute to its cardioprotective effect in ischemic models. PMID- 17132815 TI - SR 16435 [1-(1-(bicyclo[3.3.1]nonan-9-yl)piperidin-4-yl)indolin-2-one], a novel mixed nociceptin/orphanin FQ/mu-opioid receptor partial agonist: analgesic and rewarding properties in mice. AB - We identified a novel nociceptin/orphanin FQ (NOP)/mu-opioid receptor agonist, SR 16435 [1-(1-(bicyclo[3.3.1]nonan-9-yl)piperidin-4-yl)indolin-2-one], with high binding affinity and partial agonist activity at both receptors. It was hypothesized that SR 16435 would produce antinociception and yet, unlike morphine, would have diminished rewarding properties and tolerance development. Antinociception was assessed in mice using the tail-flick assay, whereas behavioral and rewarding effects were assessed using the place conditioning (PC) paradigm. PC was established by pairing drug injections with a distinct compartment. Behavioral effects were measured after acute and repeated drug administration, and the test for PC was carried out 24 h after four drug- and vehicle-pairing sessions. SR 16435 produced an increase in tail-flick latency, but SR 16435-induced antinociception was lower than that observed with morphine. Given that naloxone blocked SR 16435-induced antinociception, it is highly likely that this effect was mediated by mu-opioid receptors. Compared with morphine, chronic SR 16435 treatment resulted in reduced development of tolerance to its antinociceptive effects. SR 16435-induced conditioned place preference (CPP) was evident, an effect that was probably mediated via mu-opioid receptors, as it was reversed by coadministration of naloxone. NOP agonist activity was also present, given that SR 16435 decreased global activity, and this effect was partially reversed with the selective NOP antagonist, SR 16430 [1-(cyclooctylmethyl)-4-(3 (trifluoromethyl)phenyl)piperidin-4-ol]. Naloxone, however, also reversed the SR 16435-induced decrease in activity, indicating that both opioid and NOP receptors mediate this behavior. In summary, the mixed NOP/mu-opioid partial agonist SR 16435 exhibited both NOP and mu-opioid receptor-mediated behaviors. PMID- 17132816 TI - Potent inhibition of arterial smooth muscle tonic contractions by the selective myosin II inhibitor, blebbistatin. AB - Blebbistatin is reported to be a selective and specific small molecule inhibitor of the myosin II isoforms expressed by striated muscles and nonmuscle (IC(50) = 0.5-5 microM) but is a poor inhibitor of purified turkey smooth muscle myosin II (IC(50) approximately 80 microM). We found that blebbistatin potently (IC(50) approximately 3 microM) inhibited the actomyosin ATPase activities of expressed "slow" [smooth muscle myosin IIA (SMA)] and "fast" [smooth muscle myosin IIB (SMB)] smooth muscle myosin II heavy-chain isoforms. Blebbistatin also inhibited the KCl-induced tonic contractions produced by rabbit femoral and renal arteries that express primarily SMA and the weaker tonic contraction produced by the saphenous artery that expresses primarily SMB, with an equivalent potency comparable with that identified for nonmuscle myosin IIA (IC(50) approximately 5 microM). In femoral and saphenous arteries, blebbistatin had no effect on unloaded shortening velocity or the tonic increase in myosin light-chain phosphorylation produced by KCl but potently inhibited beta-escin permeabilized artery contracted with calcium at pCa 5, suggesting that cell signaling events upstream from KCl-induced activation of cross-bridges were unaffected by blebbistatin. It is noteworthy that KCl-induced contractions of chicken gizzard were less potently inhibited (IC(50) approximately 20 microM). Adult femoral, renal, and saphenous arteries did not express significant levels of nonmuscle myosin. These data together indicate that blebbistatin is a potent inhibitor of smooth muscle myosin II, supporting the hypothesis that the force-bearing structure responsible for tonic force maintenance in adult mammalian vascular smooth muscle is the cross-bridge formed from the blebbistatin-dependent interaction between actin and smooth muscle myosin II. PMID- 17132817 TI - Characterization of circulatory disorders in beta-thalassemic mice by noninvasive ultrasound biomicroscopy. AB - Beta-thalassemia is an inherited hematological disease caused by a decrease or absence of production of beta-globin that requires chronic therapeutic interventions. This condition leads to important arterial and venous thromboembolic events, transitory ischemic attacks, and microcirculatory obstructions, indicative of circulatory disturbances. To investigate the presence of microcirculatory disorders without the confounding effect of treatments, we used beta-thalassemic mice with typical clinical characteristics of human beta thalassemia major. One impediment to the understanding of microcirculatory physiology, in particular for beta-thalassemic mice, has been the lack of an appropriate noninvasive imaging approach. We thus developed a novel noninvasive high-frequency ultrasound imaging method to evaluate murine vascular hemodynamic properties. In our beta-thalassemic mice, total peripheral vascular resistance was significantly increased (P < 0.01) compared with wildtype littermates, whereas mean blood pressure, heart rate, and cardiac output were similar (P = nonsignificant). Importantly, the vascular hemodynamics in beta-thalassemic mice were significantly affected according to the Pourcelot indexes measured in the common carotid artery and abdominal aorta (P < 0.01 and P < 0.05, respectively). Hence, our beta-thalassemia characterization of vascular hemodynamics by noninvasive ultrasonic approaches proves the existence and provides unique quantitative assessment of microcirculatory flow disturbances in those mice. PMID- 17132818 TI - Population and systems genetics analyses of cortisol in pigs divergently selected for stress. AB - This study presents a systems genetic analysis on the physiology of cortisol in mice and pigs with an aim to show the potential of a comprehensive computational approach to quickly identify candidate genes and avoid a costly whole-genome quantitative trait locus (QTL) mapping. Population genetics analyses were performed on measurements of cortisol from a pig selection experiment. Expression QTL were mapped and gene networks were built using gene expressions for Crhr1 (corticotrophin-releasing hormone receptor) gene and single nucleotide polymorphisms from public mouse data. Results from mouse data were used to infer potential candidate regulatory genes involved in pig cortisol regulation, using a comparative or translational systems genetics approach. The pig data used were from a 10-yr divergent genetic selection experiment, providing data on 417 individuals. Population genetics analysis showed that cortisol is highly genetically determined with heritabilities of 0.40-0.70. Furthermore, a major gene with an additive effect of 86 ng/ml is segregating. Genetical-genomics investigations revealed two trans-acting eQTL for Crhr1 gene expression on chromosomes 2 and 13. Candidate gene search under trans-eQTL peaks yielded 63 genes for Crhr1 expression phenotypes. Functional links for Crhr1 genes with other genes/proteins in the gene network using mouse data were shown for the first 10 statistically significant genes involved. Results show translational or comparative systems genetics approaches reduce costs and time in large-scale genetics and "-omics" investigations. This is the first study to report a strong genetic basis for cortisol physiology using a systems approach. PMID- 17132819 TI - Aspirin activates the NF-kappaB signalling pathway and induces apoptosis in intestinal neoplasia in two in vivo models of human colorectal cancer. AB - Substantial evidence indicates that aspirin has antitumour activity against large bowel cancer and modulation of the NF-kappaB (NF-kappaB) signalling pathway has been identified as a key mechanism in this effect. However, studies examining how aspirin affects the NF-kappaB pathway to promote apoptosis have been restricted to in vitro analysis in tissue culture systems and have produced contrasting results. Here, we employed two animal models of human colorectal cancer to determine aspirin effects on the NF-kappaB pathway in colorectal neoplasia in vivo, and the relationship of such effects to the induction of apoptosis. We demonstrate that aspirin induces phosphorylation and degradation of cytoplasmic IkappaBalpha in xenografted HT-29 tumours and in adenomas from APC(Min+/-) mice. Furthermore, we show that this response occurs in a time-dependent manner and is paralleled by nuclear translocation of p65 and caspase activation. Using high performance liquid chromatography analysis, we demonstrate that >0.5 mM salicylate levels are achievable in xenografted tumours after low-dose aspirin (40 mg/kg) treatment and that these levels, which are pharmacologically relevant to humans, are sufficient to stimulate an NF-kappaB and apoptotic response. We demonstrate that activation of the NF-kappaB pathway is associated with increased apoptosis in neoplastic epithelial cells, but found that aspirin has a minimal effect on nuclear p65 and apoptosis in normal intestinal mucosa from APC(Min+/-) mice. These in vivo findings further establish that aspirin induces activation of the NF-kappaB pathway in neoplastic epithelial cells and provide further support that this effect is important for the antitumour activity of the agent. These data have considerable relevance to cancer prevention and therapy. PMID- 17132820 TI - The inositol 1,4,5-trisphosphate receptor (IP3R) and its regulators: sometimes good and sometimes bad teamwork. AB - In both nonexcitable and excitable cells, the inositol 1,4,5-trisphosphate receptor (IP(3)R) is the primary cytosolic target responsible for the initiation of intracellular calcium (Ca(2+)) signaling. To fulfill this function, the IP(3)R depends on interaction with accessory subunits and regulatory proteins. These include proteins that reside in the lumen of the endoplasmic reticulum (ER), such as chromogranin A and B and ERp44, and cytosolic proteins, such as neuronal Ca(2+) sensor 1, huntingtin, cytochrome c, IP(3)R-binding protein released with inositol 1,4,5-trisphosphate, Homer, and 4.1N. Specific interactions between these modulatory proteins and the IP(3)R have been described, making it clear that the controlled modulation of the IP(3)R by its binding partners is necessary for physiological cell regulation. The functional coupling of these modulators with the IP(3)R can control apoptosis, intracellular pH, the initiation and regulation of neuronal Ca(2+) signaling, exocytosis, and gene expression. The pathophysiological relevance of IP(3)R modulation is apparent when the functional interaction of these proteins is enhanced or abolished by mutation or overexpression. The subsequent deregulation of the IP(3)R leads to pathological changes in Ca(2+) signaling, signal initiation, the amplitude and frequency of Ca(2+) signals, and the duration of the Ca(2+) elevation. Consequences of this deregulation include abnormal growth and apoptosis. Complex regulation of Ca(2+) signaling is required for the cell to live and function, and this difficult task can only be managed when the IP(3)R teams up and acts properly with its numerous binding partners. PMID- 17132821 TI - Transgenic overexpression of protein targeting to glycogen markedly increases adipocytic glycogen storage in mice. AB - Adipocytes express the rate-limiting enzymes required for glycogen metabolism and increase glycogen synthesis in response to insulin. However, the physiological function of adipocytic glycogen in vivo is unclear, due in part to the low absolute levels and the apparent biophysical constraints of adipocyte morphology on glycogen accumulation. To further study the regulation of glycogen metabolism in adipose tissue, transgenic mice were generated that overexpressed the protein phosphatase-1 (PP1) glycogen-targeting subunit (PTG) driven by the adipocyte fatty acid binding protein (aP2) promoter. Exogenous PTG was detected in gonadal, perirenal, and brown fat depots, but it was not detected in any other tissue examined. PTG overexpression resulted in a modest redistribution of PP1 to glycogen particles, corresponding to a threefold increase in the glycogen synthase activity ratio. Glycogen synthase protein levels were also increased twofold, resulting in a combined greater than sixfold enhancement of basal glycogen synthase specific activity. Adipocytic glycogen levels were increased 200- to 400-fold in transgenic animals, and this increase was maintained to 1 yr of age. In contrast, lipid metabolism in transgenic adipose tissue was not significantly altered, as assessed by lipogenic rates, weight gain on normal or high-fat diets, or circulating free fatty acid levels after a fast. However, circulating and adipocytic leptin levels were doubled in transgenic animals, whereas adiponectin expression was unchanged. Cumulatively, these data indicate that murine adipocytes are capable of storing far higher levels of glycogen than previously reported. Furthermore, these results were obtained by overexpression of an endogenous adipocytic protein, suggesting that mechanisms may exist in vivo to maintain adipocytic glycogen storage at a physiological set point. PMID- 17132822 TI - Human brain glycogen content and metabolism: implications on its role in brain energy metabolism. AB - The adult brain relies on glucose for its energy needs and stores it in the form of glycogen, primarily in astrocytes. Animal and culture studies indicate that brain glycogen may support neuronal function when the glucose supply from the blood is inadequate and/or during neuronal activation. However, the concentration of glycogen and rates of its metabolism in the human brain are unknown. We used in vivo localized 13C-NMR spectroscopy to measure glycogen content and turnover in the human brain. Nine healthy volunteers received intravenous infusions of [1 (13)C]glucose for durations ranging from 6 to 50 h, and brain glycogen labeling and washout were measured in the occipital lobe for up to 84 h. The labeling kinetics suggest that turnover is the main mechanism of label incorporation into brain glycogen. Upon fitting a model of glycogen metabolism to the time courses of newly synthesized glycogen, human brain glycogen content was estimated at approximately 3.5 micromol/g, i.e., three- to fourfold higher than free glucose at euglycemia. Turnover of bulk brain glycogen occurred at a rate of 0.16 micromol.g-1.h-1, implying that complete turnover requires 3-5 days. Twenty minutes of visual stimulation (n=5) did not result in detectable glycogen utilization in the visual cortex, as judged from similar [13C]glycogen levels before and after stimulation. We conclude that the brain stores a substantial amount of glycogen relative to free glucose and metabolizes this store very slowly under normal physiology. PMID- 17132823 TI - Growth hormone-induced insulin resistance is associated with increased intramyocellular triglyceride content but unaltered VLDL-triglyceride kinetics. AB - The ability of growth hormone (GH) to stimulate lipolysis and cause insulin resistance in skeletal muscle may be causally linked, but the mechanisms remain obscure. We investigated the impact of GH on the turnover of FFA and VLDL-TG, intramuscular triglyceride content (IMTG), and insulin sensitivity (euglycemic clamp) in nine healthy men in a randomized double-blind placebo-controlled crossover study after 8 days treatment with (A) Placebo+Placebo, (B) GH (2 mg daily)+Placebo, and (C) GH (2 mg daily)+Acipimox (250 mgx3 daily). In the basal state, GH (B) increased FFA levels (P<0.05), palmitate turnover (P<0.05), and lipid oxidation (P=0.05), but VLDL-TG kinetics were unaffected. Administration of acipimox (C) suppressed basal lipolysis but did not influence VLDL-TG kinetics. In the basal state, IMTG content increased after GH (B; P=0.03). Insulin resistance was induced by GH irrespective of concomitant acipimox (P<0.001). The turnover of FFA and VLDL-TG was suppressed by hyperinsulinemia during placebo and GH, whereas coadministration of acipimox induced a rebound increase FFA turnover and VLDL-TG clearance. We conclude that these results show that GH-induced insulin resistance is associated with increased IMTG and unaltered VLDL-TG kinetics; we hypothesize that fat oxidation in muscle tissue is an important primary effect of GH and that circulating FFA rather than VLDL-TG constitute the major source for this process; and the role of IMTG in the development of GH induced insulin resistance merits future research. PMID- 17132824 TI - Abdominal obesity in BTBR male mice is associated with peripheral but not hepatic insulin resistance. AB - Insulin resistance is a common feature of obesity. BTBR mice have more fat mass than most other inbred mouse strains. On a chow diet, BTBR mice have elevated insulin levels relative to the C57BL/6J (B6) strain. Male F1 progeny of a B6 x BTBR cross are insulin resistant. Previously, we reported insulin resistance in isolated muscle and in isolated adipocytes in this strain. Whereas the muscle insulin resistance was observed only in male F1 mice, adipocyte insulin resistance was also present in male BTBR mice. We examined in vivo mechanisms of insulin resistance with the hyperinsulinemic euglycemic clamp technique. At 10 wk of age, BTBR and F1 mice had a >30% reduction in whole body glucose disposal primarily due to insulin resistance in heart, soleus muscle, and adipose tissue. The increased adipose tissue mass and decreased muscle mass in BTBR and F1 mice were negatively and positively correlated with whole body glucose disposal, respectively. Genes involved in focal adhesion, actin cytoskeleton, and inflammation were more highly expressed in BTBR and F1 than in B6 adipose tissue. The BTBR and F1 mice have higher levels of testosterone, which may be related to the pathological changes in adipose tissue that lead to systemic insulin resistance. Despite profound peripheral insulin resistance, BTBR and F1 mice retained hepatic insulin sensitivity. These studies reveal a genetic difference in body composition that correlates with large differences in peripheral insulin sensitivity. PMID- 17132825 TI - Hypothalamic mapping of orexigenic action and Fos-like immunoreactivity following relaxin-3 administration in male Wistar rats. AB - The insulin superfamily, characterized by common disulphide bonds, includes not only insulin but also insulin-like peptides such as relaxin-1 and relaxin-3. The actions of relaxin-3 are largely unknown, but recent work suggests a role in regulation of food intake. Relaxin-3 mRNA is highly expressed in the nucleus incertus, which has extensive projections to the hypothalamus, and relaxin immunoreactivity is present in several hypothalamic nuclei. In the rat, relaxin-3 binds and activates both relaxin family peptide receptor 1, which also binds relaxin-1, and a previously orphaned G protein-coupled receptor, RXFP3. These receptors are extensively expressed in the hypothalamus. The aims of these studies were twofold: 1) map the hypothalamic site(s) of the orexigenic action of relaxin-3 and 2) examine the site(s) of neuronal activation following central relaxin-3 administration. After microinjection into hypothalamic sites, human relaxin-3 (H3; 180 pmol) significantly stimulated 0- to 1-h food intake in the supraoptic nucleus (SON), arcuate nucleus (ARC), and the anterior preoptic area (APOA) [SON 0.4+/-0.2 (vehicle) vs. 2.9+/-0.5 g (H3), P<0.001; ARC 0.7+/-0.3 (vehicle) vs. 2.7+/-0.2 g (H3), P<0.05; and APOA 0.8+/-0.1 (vehicle) vs. 2.2+/ 0.2 g (H3), P<0.05]. Cumulative food intake was significantly increased4 mg/kg) at 35-40% in 2 wk. Animals appeared healthy during treatment and regained their normal body weights and growth trajectories upon clearance of the antibodies from the bloodstream. Measurements of food consumption and energy expenditure indicated that the rapid weight loss was induced primarily by decreased energy intake and not by increased energy expenditure or cachexia and was accompanied by a greater reduction in fat than lean body mass. Hypophagia was not caused through malaise or illness, as indicated by absence of conditioned taste aversion, pica behavior, and decreased need-induced salt intake in rats. In support of a hypothalamic site of action, we found that, after intraperitoneal injections, anti-FGFR1c (IMC-A1), but not a control antibody, accumulated in the median eminence and adjacent mediobasal hypothalamus and that FGFR1c is enriched in the hypothalamus of mice. Furthermore, a single intracerebroventricular administration of 3 microg of IMC A1 via the 3rd ventricle to mice caused an approximately 36% reduction in food intake and an approximately 6% weight loss within the ensuing 24 h. Our data suggest that FGF signaling through FGFR1c may play a physiological role in hypothalamic feeding circuit and that blocking it leads to hypophagia and weight loss. PMID- 17132827 TI - Glucagon chronically impairs hepatic and muscle glucose disposal. AB - Defects in insulin secretion and/or action contribute to the hyperglycemia of stressed and diabetic patients, and we hypothesize that failure to suppress glucagon also plays a role. We examined the chronic impact of glucagon on glucose uptake in chronically catheterized conscious depancreatized dogs placed on 5 days of nutritional support (NS). For 3 days of NS, a variable intraportal infusion of insulin was given to maintain isoglycemia (approximately 120 mg/dl). On day 3 of NS, animals received a constant low infusion of insulin (0.4 mU.kg-1.min-1) and either no glucagon (CONT), basal glucagon (0.7 ng.kg-1.min-1; BasG), or elevated glucagon (2.4 ng.kg-1.min-1; HiG) for the remaining 2 days. Glucose in NS was varied to maintain isoglycemia. An additional group (HiG+I) received elevated insulin (1 mU.kg-1.min-1) to maintain glucose requirements in the presence of elevated glucagon. On day 5 of NS, hepatic substrate balance was assessed. Insulin and glucagon levels were 10+/-2, 9+/-1, 7+/-1, and 24+/-4 microU/ml, and 24+/-5, 39+/-3, 80+/-11, and 79+/-5 pg/ml, CONT, BasG, HiG, and HiG+I, respectively. Glucagon infusion decreased the glucose requirements (9.3+/-0.1, 4.6+/-1.2, 0.9+/-0.4, and 11.3+/-1.0 mg.kg-1.min-1). Glucose uptake by both hepatic (5.1+/-0.4, 1.7+/-0.9, -1.0+/-0.4, and 1.2+/-0.4 mg.kg-1.min-1) and nonhepatic (4.2+/-0.3, 2.9+/-0.7, 1.9+/-0.3, and 10.2+/-1.0 mg.kg-1.min-1) tissues decreased. Additional insulin augmented nonhepatic glucose uptake and only partially improved hepatic glucose uptake. Thus, glucagon impaired glucose uptake by hepatic and nonhepatic tissues. Compensatory hyperinsulinemia restored nonhepatic glucose uptake and partially corrected hepatic metabolism. Thus, persistent inappropriate secretion of glucagon likely contributes to the insulin resistance and glucose intolerance observed in obese and diabetic individuals. PMID- 17132828 TI - ArrayExpress--a public database of microarray experiments and gene expression profiles. AB - ArrayExpress is a public database for high throughput functional genomics data. ArrayExpress consists of two parts--the ArrayExpress Repository, which is a MIAME supportive public archive of microarray data, and the ArrayExpress Data Warehouse, which is a database of gene expression profiles selected from the repository and consistently re-annotated. Archived experiments can be queried by experiment attributes, such as keywords, species, array platform, authors, journals or accession numbers. Gene expression profiles can be queried by gene names and properties, such as Gene Ontology terms and gene expression profiles can be visualized. ArrayExpress is a rapidly growing database, currently it contains data from >50,000 hybridizations and >1,500,000 individual expression profiles. ArrayExpress supports community standards, including MIAME, MAGE-ML and more recently the proposal for a spreadsheet based data exchange format: MAGE TAB. AVAILABILITY: www.ebi.ac.uk/arrayexpress. PMID- 17132829 TI - ECgene: an alternative splicing database update. AB - ECgene (http://genome.ewha.ac.kr/ECgene) was developed to provide functional annotation for alternatively spliced genes. The applications encompass the genome based transcript modeling for alternative splicing (AS), domain analysis with Gene Ontology (GO) annotation and expression analysis based on the EST and SAGE data. We have expanded the ECgene's AS modeling and EST clustering to nine organisms for which sufficient EST data are available in the GenBank. As for the human genome, we have also introduced several new applications to analyze differential expression. ECprofiler is an ontology-based candidate gene search system that allows users to select an arbitrary combination of gene expression pattern and GO functional categories. DEGEST is a database of differentially expressed genes and isoforms based on the EST information. Importantly, gene expression is analyzed at three distinctive levels-gene, isoform and exon levels. The user interfaces for functional and expression analyses have been substantially improved. ASviewer is a dedicated java application that visualizes the transcript structure and functional features of alternatively spliced variants. The SAGE part of the expression module provides many additional features including SNP, differential expression and alternative tag positions. PMID- 17132830 TI - GeneSpeed: protein domain organization of the transcriptome. AB - The GeneSpeed database (http://genespeed.uchsc.edu/) is an online database and resource tool facilitating the detailed study of protein domain homology in the transcriptomes of Homo sapiens, Mus musculus, Drosophila melanogaster and Caenorhabditis elegans. The population schema for the GeneSpeed database takes advantage of HOWARD parallel cluster technology (http://www.massivelyparallel.com/) and performs exhaustive tBLASTn searches covering all pre-assigned PFAM domain classes in all species (currently 7973 domain families) against the respective Unigene EST databases of the selected four transcriptomes. The resulting database provides a complete annotation of presumed protein domain presence for each Unigene cluster. To complement this domain annotation we have also performed a custom transcription factor-family curation of all Pfam domains, incorporated the Gene Ontology classifications for these domains as well as integrated the Novartis SymAtlas2 dataset for both human and mouse which provides rapid and easy access to tissue-based expression analysis. Consequently, the GeneSpeed database provides the user with the capability to browse or search the database by any of these specialized criteria as well as more traditional means (gene identifier, gene symbol, etc.), thereby enabling a supervised analysis of gene families through a top-down hierarchical basis defined by domain content, all directly linked to an optimized gene expression dataset. PMID- 17132831 TI - PlasmID: a centralized repository for plasmid clone information and distribution. AB - The Plasmid Information Database (PlasmID; http://plasmid.hms.harvard.edu) was developed as a community-based resource portal to facilitate search and request of plasmid clones shared with the Dana-Farber/Harvard Cancer Center (DF/HCC) DNA Resource Core. PlasmID serves as a central data repository and enables researchers to search the collection online using common gene names and identifiers, keywords, vector features, author names and PubMed IDs. As of October 2006, the repository contains >46 000 plasmids in 98 different vectors, including cloned cDNA and genomic fragments from 26 different species. Moreover, the clones include plasmid vectors useful for routine and cutting-edge techniques; functionally related sets of human cDNA clones; and genome-scale gene collections for Saccharomyces cerevisiae, Pseudomonas aeruginosa, Yersinia pestis, Francisella tularensis, Bacillus anthracis and Vibrio cholerae. Information about the plasmids has been fully annotated in adherence with a high quality standard, and clone samples are stored as glycerol stocks in a state-of the-art automated -80 degrees C freezer storage system. Clone replication and distribution is highly automated to minimize human error. Infor-mation about vectors and plasmid clones, including downloadable maps and sequence data, is freely available online. Researchers interested in requesting clone samples or sharing their own plasmids with the repository can visit the PlasmID website for more information. PMID- 17132832 TI - FireDB--a database of functionally important residues from proteins of known structure. AB - The FireDB database is a databank for functional information relating to proteins with known structures. It contains the most comprehensive and detailed repository of known functionally important residues, bringing together both ligand binding and catalytic residues in one site. The platform integrates biologically relevant data filtered from the close atomic contacts in Protein Data Bank crystal structures and reliably annotated catalytic residues from the Catalytic Site Atlas. The interface allows users to make queries by protein, ligand or keyword. Relevant biologically important residues are displayed in a simple and easy to read manner that allows users to assess binding site similarity across homologous proteins. Binding site residue variations can also be viewed with molecular visualization tools. The database is available at http://firedb.bioinfo.cnio.es. PMID- 17132834 TI - Psychosocial benefits of three formats of a standardized behavioral stress management program. AB - OBJECTIVE: Psychosocial factors are associated with increased morbidity and mortality in healthy and clinical populations. Behavioral interventions are needed to train the large number of people in the community setting who are affected by stressors to use coping skills that will reduce these risk factors. The aim of the current study was to evaluate the efficacy of three forms of delivery of a standardized, behavioral intervention-the Williams LifeSkills program-designed to reduce levels of psychosocial risk factors in nonclinical populations. METHODS: One hundred ninety-six participants screening positive for elevated psychosocial distress were randomized to either a waitlist control group or one of three intervention groups: the LifeSkills Workshop, the LifeSkills Video, or the LifeSkills Video and Workshop combined. Psychosocial risk factors were evaluated at baseline and at 10 days, 2 months, and 6 months after the training/wait period. RESULTS: At 10 days follow up, the workshop + video and video-only groups showed significant improvements over control subjects in trait anxiety and perceived stress. Moreover, the workshop + video group maintained benefit over control subjects throughout 6 months follow up in both of these measures, whereas the video-only group maintained benefit in trait anxiety. CONCLUSIONS: Because the psychosocial well-being of two of the treated groups improved over that of the control group, it appears that the Williams LifeSkills program accelerates and maintains a normal return to low distress after a stressful time. This is the first study to show that a commercially available, facilitator- or self-administered behavioral training product can have significant beneficial effects on psychosocial well-being in a healthy community sample. PMID- 17132835 TI - Predicting blood pressure and heart rate change with cardiovascular reactivity and recovery: results from 3-year and 10-year follow up. AB - OBJECTIVES: We examined whether cardiovascular reactivity and recovery after laboratory-induced stress is useful in predicting 3-year and 10-year ambulatory blood pressure (BP) and heart rate (HR) among 330 initially normotensive individuals. METHODS: At baseline, BP and HR measurements were recorded during three 5-minute laboratory challenges and three 5-minute recovery periods after each challenge. Measurements of systolic BP, diastolic BP, and HR were collected throughout this baseline protocol and also at 3-year and 10-year follow up by ambulatory monitoring. RESULTS: After adjustment for traditional biologic predictors, reactivity was found to explain significant variance in follow-up data across all 3-year indices and two of the 10-year indices. Recovery, entered in a following step after reactivity, was found to explain additional significant variance across all 3-year indices but none of the 10-year indices. Family hypertension history data were not found to be significantly associated with reactivity or recovery nor were they predictive of longitudinal ambulatory data after adjustment for initial resting cardiovascular levels. CONCLUSION: Overall, from a hierarchical regression model perspective, the data support the use of both reactivity and recovery in clinical predictions of proximal BP and HR and generally support the use of reactivity (but not recovery) in long-term BP predictions. PMID- 17132836 TI - Anticipatory cognitive stress appraisal and the acute procoagulant stress response in men. AB - OBJECTIVE: Acute mental stress elicits blood hypercoagulability. Following a transactional stress model, we investigated whether individuals who anticipate stress as more threatening, challenging, and as exceeding their coping skills show greater stress reactivity of the coagulation activation marker D-dimer, indicating fibrin generation in plasma. METHODS: Forty-seven men (mean age 44 +/- 14 years; mean blood pressure [MBP] 101 +/- 12 mm Hg; mean body mass index [BMI] 26 +/- 3 kg/m(2)) completed the Primary Appraisal Secondary Appraisal (PASA) scale before undergoing the Trier Social Stress Test (combination of mock job interview and mental arithmetic task). Heart rate, blood pressure, plasma catecholamines, and D-dimer levels were measured before and after stress, and during recovery up to 60 minutes poststress. RESULTS: Hemodynamic measures, catecholamines, and D-dimer changed across all time points (p values <.001). The PASA "Stress Index" (integrated measure of transactional stress perception) correlated with total D-dimer area under the curve (AUC) between rest and 60 minutes poststress (r = 0.30, p = .050) and with D-dimer change from rest to immediately poststress (r = 0.29, p = .046). Primary appraisal (combined "threat" and "challenge") correlated with total D-dimer AUC (r = 0.37, p = .017), D-dimer stress change (r = 0.41, p = .004), and D-dimer recovery (r = 0.32, p = .042). "Challenge" correlated more strongly with D-dimer stress change than "threat" (p = .020). Primary appraisal (DeltaR(2) = 0.098, beta = 0.37, p = .019), and particularly its subscale "challenge" (DeltaR(2) = 0.138, beta = 0.40, p = .005), predicted D-dimer stress change independently of age, BP, BMI, and catecholamine change. CONCLUSIONS: Anticipatory cognitive appraisal determined the extent of coagulation activation to and recovery from stress in men. Particularly individuals who anticipated the stressor as more challenging and also more threatening had a greater fibrin stress response. PMID- 17132837 TI - Burnout and risk of type 2 diabetes: a prospective study of apparently healthy employed persons. AB - OBJECTIVE: This prospective study was designed to test the extent to which the onset of type 2 diabetes in apparently healthy individuals was predicted by burnout, a unique affective response to combined exposure to chronic stressors. METHODS: The study participants were 677 employed men and women who were followed up for 3 to 5 years (mean = 3.6 years) for the onset of diagnosed type 2 diabetes. Burnout was assessed by the Shirom-Melamed Burnout Measure with its three subscales: emotional exhaustion, physical fatigue, and cognitive weariness. RESULTS: The burnout symptoms were remarkably consistent over the follow-up period irrespective of changes in place of work and in employment status. During the follow-up period, 17 workers developed type 2 diabetes. Logistic regression results indicated that burnout was associated with a 1.84-fold increased risk of diabetes (95% confidence interval [CI] = 1.19-2.85) even after adjusting for age, sex, body mass index, smoking, alcohol use, leisure time physical activity, initial job category, and follow-up duration. In a subsample of 507 workers, the relative risk of diabetes was found to be much higher after additional control for blood pressure levels (odds ratio = 4.32, 95% CI = 1.75-10.67), available only for this subsample. CONCLUSIONS: These findings suggest that chronic burnout might be a risk factor for the onset of type 2 diabetes in apparently healthy individuals. PMID- 17132838 TI - Daily goal pursuits predict cortisol secretion and mood states in employed parents with preschool children. AB - OBJECTIVE: This study examined the relationship between the personal relevance of daily activities with respect to self-set work and family goals and affective and neuroendocrine stress reactions. METHODS: A total of 53 dual-earner couples with preschool children participated in a 1-week interval-sampling study. At the beginning, participants reported their personal work and family goals. During the time-sampling phase, both partners reported the goal relevance of their daily activities, current mood, and provided saliva samples for cortisol estimation every 3 hours. RESULTS: Hierarchical linear models show that the performance of goal-furthering activities is associated with more positive mood and decreased secretion of cortisol. The relationship between the goal relevance of daily activities and cortisol was partially mediated by affect quality. CONCLUSIONS: These findings speak to a person-centered approach in research on stress by showing that knowledge of individual goals is important for an understanding of affective and neuroendocrine stress reactions in employed parents with preschool children. PMID- 17132839 TI - Interleukin-6 covaries inversely with cognitive performance among middle-aged community volunteers. AB - OBJECTIVE: Recent evidence suggests that higher peripheral levels of interleukin 6 (IL-6) are associated with poorer cognitive function and predict future cognitive decline among the elderly. The current investigation extends the study of relationships between plasma IL-6 and cognitive performance to healthy middle aged adults and to an examination of more specific cognitive domains. METHODS: Five hundred relatively healthy community volunteers aged 30 to 54 had blood drawn for the determination of plasma IL-6 levels and completed a battery of neuropsychological tests evaluating memory and executive function. RESULTS: After controlling for age, gender, race, and education, hierarchical regression analyses revealed an inverse relationship between circulating levels of IL-6 and performance on clusters of tests assessing auditory recognition memory, attention/working memory, and executive function. In contrast, there was no association between IL-6 and performance on tests of general memory. Secondary analyses demonstrated that relationships between IL-6 and auditory recognition and working memory and executive function were independent of a number of health factors, including body mass index, smoking, and hypertension. CONCLUSIONS: These findings contribute to a growing body of evidence linking chronic inflammation to poorer cognitive functioning and extend these findings to a midlife community sample, raising the possibility that IL-6 may represent a biomarker for risk of future cognitive decline. PMID- 17132840 TI - Are stress-induced cortisol changes during pregnancy associated with postpartum depressive symptoms? AB - OBJECTIVE: The purpose of this study was to examine the association between psychobiological stress reactivity during healthy pregnancy and depressive symptoms in the early puerperium. METHODS: A sample of healthy nulliparous pregnant women (N = 57) between the ages of 21 and 35 years underwent a standardized psychosocial stress test during pregnancy. Within an average of 13 days after delivery, postpartum depressive symptoms were assessed using the German version of the Edinburgh postnatal depression scale (EPDS). The sample was divided into a group with probable cases (EPDS score >9, N = 16) and a group with probable noncases (EPDS score < or =9, N = 41). RESULTS: The probable case group showed significantly higher cortisol responses to the stress test compared with the probable noncase group, whereas baseline levels did not differ. Additionally, women in the probable case group showed significantly higher state anxiety and lower mood state throughout the experiment. Furthermore, the probable case group showed higher stress susceptibility, higher trait anxiety, and higher levels in the Symptom Checklist. No differences were found for prior episodes of psychiatric disorders, obstetrical complications, birth weight, or mode of delivery. CONCLUSIONS: Our data provide evidence that healthy pregnant women developing postpartum depressive symptoms might already be identified during pregnancy by means of their higher cortisol reactivity and their higher psychological reactivity in response to psychosocial stress. Further investigations are required to explore whether higher psychobiological stress responses not only precede depressive symptoms within 2 weeks after birth, but might also predict postpartum major depression. PMID- 17132841 TI - The asthma trigger inventory: validation of a questionnaire for perceived triggers of asthma. AB - BACKGROUND: Asthma patients' perceptions of triggers have been explored in a largely unstructured fashion in the past. We therefore developed and validated a questionnaire of commonly perceived asthma triggers. METHODS: Two hundred forty seven primary care patients with asthma filled in an asthma trigger survey together with questionnaires on demographics, asthma-relevant information, perceived control of asthma, and general health status. Factor structure of the item pool and psychometric properties of trigger subscales were evaluated. We also investigated the relationship between allergen or psychological trigger reports and allergy skin test response or respiratory impedance during emotional film viewing, respectively. RESULTS: Principal component analysis yielded six factors that were thematically associated with psychology, animal allergens, pollen allergens, physical activity, infection, and air pollution/irritants. Subscales showed good internal consistencies and low to moderately positive intercorrelations. Psychological triggers were consistently associated with less favorable health status, a reduced perception of asthma control, and greater medical treatment utilization. Animal allergen scores correlated positively with skin test responses to animal allergens. Respiratory impedance increases during emotional film clips were positively correlated with the psychological trigger subscale. CONCLUSION: The questionnaire is a reliable measure of commonly perceived asthma triggers. Aspects of patients' trigger reports reflect actual reactivity to specific trigger factors. PMID- 17132842 TI - Temporal factors alter effects of social housing conditions on responses to chemotherapy and hormone levels in a Shionogi mammary tumor model. AB - OBJECTIVE: To identify possible hormonal factors involved in the differential responses to chemotherapy observed in our tumor model, we investigated if the timing among tumor cell injection, rehousing, and chemotherapy administration differentially affects levels of corticosterone (CORT), growth hormone (GH), and testosterone and tumor and host responses to chemotherapy. METHODS: Mice were reared either individually (I) or in groups (G). At 2 to 4 months, mice were injected with tumor cells and retained in their original housing conditions or rehoused into different experimental groups (GG, IG, II, GI) either immediately (experiment 1) or 14 days later (experiment 2); chemotherapy was administered when tumors weighed approximately 0.8 g. RESULTS: In experiment 1, IG and GG mice had better responses to chemotherapy than GI mice. Chemotherapy increased CORT levels in II mice and decreased GH levels in GI mice compared with those of their drug vehicle-treated counterparts. Under the temporal conditions of experiment 2, IG and GG mice lost the advantage seen in experiment 1 in terms of tumor and host responses to chemotherapy. Before chemotherapy administration, CORT levels in IG mice and GH levels in GI mice were higher than those in mice in all other housing conditions. At 1 day after chemotherapy, CORT levels were higher for chemotherapy treated than for drug vehicle-treated IG mice, and at 5 days post chemotherapy, GH levels were higher in GI than in IG mice. CONCLUSIONS: Temporal relationships among tumor cell injection, rehousing, and chemotherapy administration critically influence responses to chemotherapy; these effects may be mediated, in part, by alterations in hormone levels. PMID- 17132843 TI - Health problems and medical utilization associated with gambling disorders: results from the National Epidemiologic Survey on Alcohol and Related Conditions. AB - OBJECTIVE: Pathologic gambling is believed to be associated with adverse health consequences, but no prior studies have rigorously evaluated these relationships. We sought to examine medical disorders and health service utilization associated with problem and pathologic gambling. METHOD: A total of 43,093 adults aged 18 years and older were evaluated in the 2001 to 2002 National Epidemiologic Survey on Alcohol and Related Conditions. Self-reported medical diagnoses and past-year medical services used were assessed. RESULTS: Pathologic gamblers were more likely than low-risk individuals to have been diagnosed with tachycardia (odds ratio [OR] = 1.77; 95% confidence interval [CI] = 1.05-2.97), angina (OR = 2.35; 95% CI = 1.33-4.15), cirrhosis (OR = 3.90; 95% CI = 1.11-13.72), and other liver disease (OR = 2.98; 95% CI = 1.07-8.26). Gambling severity was also associated with higher rates of medical utilization with pathologic gamblers more likely than low-risk individuals to have been treated in the emergency room in the year before the survey (OR = 1.98; 95% CI = 1.27-3.09). Significant effects of gambling severity remained even after controlling for demographic characteristics (age, gender, ethnicity, marital status, education, income, and region of the country) and behavioral risk factors such as body mass index, alcohol abuse and dependence, nicotine dependence, and mood and anxiety disorders. CONCLUSIONS: A lifetime diagnosis of pathologic gambling is associated with several medical disorders and increased medical utilization, perhaps leading to a burden on healthcare costs in the United States. PMID- 17132844 TI - Coping and burden among informal HIV caregivers. AB - OBJECTIVE: The aim of this study was to examine the role of coping on caregiver burden among a heterogeneous group of caregivers of persons living with HIV during the era of highly active antiretroviral therapy. METHODS: Burden and coping were examined among 176 caregivers of persons living with HIV. Three styles of coping were examined using a 7-item scale: active-approach (task), blame-withdrawal (emotion), and distancing (avoidance). RESULTS: A total of 58.8% of the caregivers were women. They had a mean age of 42 years; 61.9% cohabited with the persons living with HIV who had a mean CD4 count of 401. All three styles of coping were significantly positively correlated with caregiver burden. After controlling for demographic variables and caregiver depression, active approach coping and distancing coping independently moderated the relationship between perceived severity of HIV-related symptoms (stress) and caregiver burden; however, some caregivers experienced burden even at low levels of stress. CONCLUSIONS: These results indicate that in the era of highly active antiretroviral therapy, coping mitigates the effect of stress on burden. PMID- 17132845 TI - Patients do not demand and doctors do not misunderstand--how medically unexplained symptoms become medicalized. PMID- 17132848 TI - Overview of nomenclature of nuclear receptors. AB - Nuclear receptor pharmacology has, to a certain extent, led the way, compared with other receptor systems, in the appreciation that ligands may exert very diverse pharmacology, based on their individual chemical structure and the allosteric changes induced in the receptor/accessory protein complex. This can lead to very selective pharmacological effects, which may not necessarily be predicted from the experience with other agonists/partial agonists/antagonists. If this is the case, then drug discovery may be back to drug-specific pharmacology (where each drug may have an original profile), rather than specific drug pharmacology (where agents specific for a receptor have a distinct profile). As functional selectivity is indeed a crucial mechanism to be considered when going through the drug discovery development process, then initial screens using reconstituted systems may not show the appropriate pharmacology, simply because the required stoichiometry of corepressors and coactivators may not be present to select the best compounds; therefore, multiple effector systems are necessary to screen for differential activation, and, even then, screening with in vivo pathophysiological models may ultimately be required for the selection process-a massive but necessary task for pharmacologists. Thus, the characterization of nuclear receptors and their associated proteins and the ligands that interact with them will remain a challenge to pharmacologists. PMID- 17132849 TI - International Union of Pharmacology. LIX. The pharmacology and classification of the nuclear receptor superfamily: thyroid hormone receptors. PMID- 17132850 TI - International Union of Pharmacology. LX. Retinoic acid receptors. AB - Retinoid is a term for compounds that bind to and activate retinoic acid receptors (RARalpha, RARbeta, and RARgamma), members of the nuclear hormone receptor superfamily. The most important endogenous retinoid is all-trans retinoic acid. Retinoids regulate a wide variety of essential biological processes, such as vertebrate embryonic morphogenesis and organogenesis, cell growth arrest, differentiation and apoptosis, and homeostasis, as well as their disorders. This review summarizes the considerable amount of knowledge generated on these receptors. PMID- 17132851 TI - International Union of Pharmacology. LXI. Peroxisome proliferator-activated receptors. AB - The three peroxisome proliferator-activated receptors (PPARs) are ligand activated transcription factors of the nuclear hormone receptor superfamily. They share a high degree of structural homology with all members of the superfamily, particularly in the DNA-binding domain and ligand- and cofactor-binding domain. Many cellular and systemic roles have been attributed to these receptors, reaching far beyond the stimulation of peroxisome proliferation in rodents after which they were initially named. PPARs exhibit broad, isotype-specific tissue expression patterns. PPARalpha is expressed at high levels in organs with significant catabolism of fatty acids. PPARbeta/delta has the broadest expression pattern, and the levels of expression in certain tissues depend on the extent of cell proliferation and differentiation. PPARgamma is expressed as two isoforms, of which PPARgamma2 is found at high levels in the adipose tissues, whereas PPARgamma1 has a broader expression pattern. Transcriptional regulation by PPARs requires heterodimerization with the retinoid X receptor (RXR). When activated by a ligand, the dimer modulates transcription via binding to a specific DNA sequence element called a peroxisome proliferator response element (PPRE) in the promoter region of target genes. A wide variety of natural or synthetic compounds was identified as PPAR ligands. Among the synthetic ligands, the lipid-lowering drugs, fibrates, and the insulin sensitizers, thiazolidinediones, are PPARalpha and PPARgamma agonists, respectively, which underscores the important role of PPARs as therapeutic targets. Transcriptional control by PPAR/RXR heterodimers also requires interaction with coregulator complexes. Thus, selective action of PPARs in vivo results from the interplay at a given time point between expression levels of each of the three PPAR and RXR isotypes, affinity for a specific promoter PPRE, and ligand and cofactor availabilities. PMID- 17132852 TI - International Union of Pharmacology. LXII. The NR1H and NR1I receptors: constitutive androstane receptor, pregnene X receptor, farnesoid X receptor alpha, farnesoid X receptor beta, liver X receptor alpha, liver X receptor beta, and vitamin D receptor. AB - The nuclear receptors of the NR1H and NR1I subgroups include the constitutive androstane receptor, pregnane X receptor, farnesoid X receptors, liver X receptors, and vitamin D receptor. The newly emerging functions of these related receptors are under the control of metabolic pathways, including metabolism of xenobiotics, bile acids, cholesterol, and calcium. This review summarizes results of structural, pharmacologic, and genetic studies of these receptors. PMID- 17132853 TI - International Union of Pharmacology. LXIII. Retinoid X receptors. AB - The physiological effects of retinoic acids (RAs) are mediated by members of two families of nuclear receptors, the retinoic acid receptors (RARs) and the retinoid X receptors (RXRs), which are encoded by three distinct human genes, RXRalpha, RXRbeta, and RXRgamma. RARs bind both all-trans- and 9-cis-RA, whereas only the 9-cis-RA stereoisomer binds to RXRs. As RXR/RAR heterodimers, these receptors control the transcription of RA target genes through binding to RA response elements. This review is focused on the structure, mode of action, ligands, expression, and pharmacology of RXRs. Given their role as common partners to many other members of the nuclear receptor superfamily, these receptors have been the subject of intense scrutiny. Moreover, and despite numerous studies since their initial discovery, RXRs remain enigmatic nuclear receptors, and there is still no consensus regarding their role. Indeed, multiple questions about the actual biological role of RXRs and the existence of an endogenous ligand have still to be answered. PMID- 17132854 TI - International Union of Pharmacology. LXIV. Estrogen receptors. PMID- 17132855 TI - International Union of Pharmacology. LXV. The pharmacology and classification of the nuclear receptor superfamily: glucocorticoid, mineralocorticoid, progesterone, and androgen receptors. PMID- 17132856 TI - International Union of Pharmacology. LXVI. Orphan nuclear receptors. AB - Half of the members of the nuclear receptors superfamily are so-called "orphan" receptors because the identity of their ligand, if any, is unknown. Because of their important biological roles, the study of orphan receptors has attracted much attention recently and has resulted in rapid advances that have helped in the discovery of novel signaling pathways. In this review we present the main features of orphan receptors, discuss the structure of their ligand-binding domains and their biological functions. The paradoxical existence of a pharmacology of orphan receptors, a rapidly growing and innovative field, is highlighted. PMID- 17132857 TI - Direct G protein modulation of Cav2 calcium channels. AB - The regulation of presynaptic, voltage-gated calcium channels by activation of heptahelical G protein-coupled receptors exerts a crucial influence on presynaptic calcium entry and hence on neurotransmitter release. Receptor activation subjects presynaptic N- and P/Q-type calcium channels to a rapid, membrane-delimited inhibition-mediated by direct, voltage-dependent interactions between G protein betagamma subunits and the channels-and to a slower, voltage independent modulation involving soluble second messenger molecules. In turn, the direct inhibition of the channels is regulated as a function of many factors, including channel subtype, ancillary calcium channel subunits, and the types of G proteins and G protein regulatory factors involved. Twenty-five years after this mode of physiological regulation was first described, we review the investigations that have led to our current understanding of its molecular mechanisms. PMID- 17132858 TI - When worlds colloid. PMID- 17132859 TI - Identifying protein construct variants with increased crystallization propensity- a case study. AB - This study describes an efficient multiparallel automated workflow of cloning, expression, purification, and crystallization of a large set of construct variants for isolated protein domains aimed at structure determination by X-ray crystallography. This methodology is applied to MAPKAP kinase 2, a key enzyme in the inflammation pathway and thus an attractive drug target. The study reveals a distinct subset of truncation variants with improved crystallization properties. These constructs distinguish themselves by increased solubility and stability during a parallel automated multistep purification process including removal of the recombinant tag. High-throughput protein melting point analysis characterizes this subset of constructs as particularly thermostable. Both parallel purification screening and melting point determination clearly identify residue 364 as the optimal C terminus for the kinase domain. Moreover, all three constructs that ultimately crystallized feature this C terminus. At the N terminus, only three amino acids differentiate a noncrystallizing from a crystallizing construct. This study addresses the very common issues associated with difficult to crystallize proteins, those of solubility and stability, and the crucial importance of particular residues in the formation of crystal contacts. A methodology is suggested that includes biophysical measurements to efficiently identify and produce construct variants of isolated protein domains which exhibit higher crystallization propensity. PMID- 17132860 TI - Site-directed mutagenesis indicates an important role of cysteines 76 and 181 in the catalysis of hydantoin racemase from Sinorhizobium meliloti. AB - Hydantoin racemase enzyme plays a crucial role in the reaction cascade known as "hydantoinase process." In conjunction with a stereoselective hydantoinase and a stereospecific carbamoylase, it allows the total conversion from D,L-5 monosubstituted hydantoins, with a low rate of racemization, to optically pure D- or L-amino acids. Residues Cys76 and Cys181 belonging to hydantoin racemase from Sinorhizobium meliloti (SmeHyuA) have been proved to be involved in catalysis. Here, we report biophysical data of SmeHyuA Cys76 and Cys181 to alanine mutants, which point toward a two-base mechanism for the racemization of 5-monosubstituted hydantoins. The secondary and the tertiary structure of the mutants were not significantly affected, as shown by circular dichroism. Calorimetric and fluorescence experiments have shown that Cys76 is responsible for recognition and proton retrieval of D-isomers, while Cys181 is responsible for L-isomer recognition and racemization. This recognition process is further supported by measurements of protein stability followed by chemical denaturation in the presence of the corresponding compound. PMID- 17132861 TI - Critical assessment of quantum mechanics based energy restraints in protein crystal structure refinement. AB - A critical evaluation of the performance of X-ray refinement protocols using various energy functions is presented using the bovine pancreatic trypsin inhibitor (BPTI) protein. The four potential energy functions we explored include: (1) fully quantum mechanical calculations; (2) one based on an incomplete molecular mechanics (MM) energy function employed in the Crystallography and NMR System (CNS) with empirical parameters developed by Engh and Huber (EH), which lacks electrostatic and attractive van der Waals terms; (3) one based on a complete MM energy function (AMBER ff99 parameter set); and (4) the same as 3, with the addition of a Generalized Born (GB) implicit solvation term. The R, R (free), real space R values of the refined structures and deviations from the original experimental structure were used to assess the relative performance. It was found that at 1 Angstrom resolution the physically based energy functions 1, 3, and 4 performed better than energy function 2, which we attribute to the better representation of key interactions, particularly electrostatics. The observed departures from the experimental structure were similar for the refinements with physically based energy functions and were smaller than the structure refined with EH. A test refinement was also performed with the reflections truncated at a high-resolution cutoff of 2.5 Angstrom and with random perturbations introduced into the initial coordinates, which showed that low-resolution refinements with physically based energy functions held the structure closer to the experimental structure solved at 1 Angstrom resolution than the EH-based refinements. PMID- 17132862 TI - New algorithms and an in silico benchmark for computational enzyme design. AB - The creation of novel enzymes capable of catalyzing any desired chemical reaction is a grand challenge for computational protein design. Here we describe two new algorithms for enzyme design that employ hashing techniques to allow searching through large numbers of protein scaffolds for optimal catalytic site placement. We also describe an in silico benchmark, based on the recapitulation of the active sites of native enzymes, that allows rapid evaluation and testing of enzyme design methodologies. In the benchmark test, which consists of designing sites for each of 10 different chemical reactions in backbone scaffolds derived from 10 enzymes catalyzing the reactions, the new methods succeed in identifying the native site in the native scaffold and ranking it within the top five designs for six of the 10 reactions. The new methods can be directly applied to the design of new enzymes, and the benchmark provides a powerful in silico test for guiding improvements in computational enzyme design. PMID- 17132863 TI - Specific interactions by the N-terminal arm inhibit self-association of the AraC dimerization domain. AB - Deletion of the regulatory N-terminal arms of the AraC protein from its dimerization domain fragments increases the susceptibility of the dimerization domain to form a series of higher order polymers by indefinite self-association. We investigated how the normal presence of the arm inhibits this self association. One possibility is that arms can act as an entropic bristles to interfere with the approach of other macromolecules, thereby decreasing collision frequencies. We examined the repulsive effect of flexible arms by measuring the rate of trypsin cleavage of a specially constructed ubiquitin-arm protein. Adding an arm to ubiquitin or increasing its length produced only a modest repulsive effect. This suggests that arms such as the N-terminal arm of AraC do not reduce self-association by entropic exclusion. We consequently tested the hypothesis that the arm on AraC reduces self-association by binding to the core of the dimerization domain even in the absence of arabinose. The behaviors of dimerization domain mutants containing deletions or alterations in the N-terminal arms substantiate this hypothesis. Apparently, interactions between the N terminal arm and the dimerization domain core position the arm to interfere with the protein-protein contacts necessary for self-association. PMID- 17132864 TI - Enhanced cell-free protein expression by fusion with immunoglobulin Ckappa domain. AB - While cell-free systems are increasingly used for protein expression in structural and functional studies, several proteins are difficult to express or expressed only at low levels in cell-free lysates. Here, we report that fusion of the human immunoglobulin kappa light chain constant domain (Ckappa) at the C terminus of four representative proteins dramatically improved their production in the Escherichia coli S30 system, suggesting that enhancement of cell-free protein expression by Ckappa fusion will be widely applicable. PMID- 17132865 TI - Identification and characterization of human ethanolaminephosphotransferase1. AB - CDP-ethanolamine:diacylglycerol ethanolaminephosphotransferase (EPT) catalyzes the transfer of phosphoethanolamine from CDP-ethanolamine to diacylglycerol to produce phosphatidylethanolamine (PE). To date, the dual specificity of choline/ethanolaminephosphotransferase (CEPT) has been recognized as the total activity responsible for the synthesis of PE via the CDP-ethanolamine pathway in human. We report here the identification and characterization of another human cDNA that encodes CDP-ethanolamine-specific human EPT (hEPT1). Through homology search, we found that human selenoprotein I contained the CDP-alcohol phosphatidyltransferase signature, a common motif conserved in phospholipid synthases. Bacterial expression of the cDNA in Escherichia coli demonstrated that the product specifically used CDP-ethanolamine as the phosphobase donor to produce PE with the activation by both Mn(2+) and Mg(2+). RT-PCR and Northern blot analysis revealed that hEPT1 was ubiquitously expressed in multiple tissues, but in brain it was highly expressed in cerebellum. Here, we propose that in addition to previously identified CEPT, hEPT1 is involved in the biosynthesis of PE via the Kennedy pathway. PMID- 17132866 TI - Glomerular and tubular function during AT1 receptor blockade in pigs with neonatal induced partial ureteropelvic obstruction. AB - Previously, we showed that neonatal induced chronic partial unilateral ureteral obstruction (PUUO) of the multipapillary pig kidney decreased glomerular filtration rate (GFR) of the obstructed kidney. We hypothesized that ANG II and nitric oxide (NO) are important for the changes in renal function and in the present study we examined the effects of chronic AT1 receptor blockade using CV 11974 (0.12 mg/h candesartan from age 23 to 30 days) on kidney function development after PUUO was induced in 2-day-old piglets. Moreover, the effect of superimposed acute NO inhibition using N(G)-nitro-l-arginine methyl ester (l NAME; 15 mg/kg) was examined to identify if this has diagnostic potential. PUUO significantly increased GFR in the nonobstructed contralateral kidney independent of candesartan. In candesartan-treated piglets, the l-NAME-induced GFR reduction seen in normal and nonobstructed kidneys was absent in the partial obstructed kidneys. Urine output and fractional excretion of water were increased from the partial obstructed kidneys. Consistent with this immunohistochemical analyses showed a reduced aquaporin-2 labeling in the collecting duct principal cells. Moreover, renal sodium handling was compromised by PUUO evidenced by an increased fractional excretion of sodium which was enhanced by candesartan treatment. In conclusion, our findings suggest that the counterbalance between AT1 receptor mediated vasoconstriction and NO-mediated vasodilatation which maintain GFR in normal young porcine kidneys is changed by neonatal induced chronic PUUO. This may have diagnostic potential in children with suspected congenital obstruction. Our results also demonstrate compromised tubular functions in response to chronic PUUO despite preservation of glomerular function. PMID- 17132867 TI - Flow increases superoxide production by NADPH oxidase via activation of Na-K-2Cl cotransport and mechanical stress in thick ascending limbs. AB - Superoxide (O(2)(-)) regulates renal function and is implicated in hypertension. O(2)(-) production increases in response to increased ion delivery in thick ascending limbs (TALs) and macula densa and mechanical strain in other cell types. Tubular flow in the kidney acutely varies causing changes in ion delivery and mechanical stress. We hypothesized that increasing luminal flow stimulates O(2)(-) production by NADPH oxidase in TALs via activation of Na-K-2Cl cotransport. We measured intracellular O(2)(-) in isolated rat TALs using dihydroethidium in the presence and absence of luminal flow and inhibitors of NADPH oxidase, Na-K-2Cl cotransport, and Na/H exchange. In the absence of flow, the rate of O(2)(-) production was 5.8 +/- 1.4 AU/s. After flow was initiated, it increased to 29.7 +/- 4.3 AU/s (P < 0.001). O(2)(-) production was linearly related to flow. Tempol alone and apocynin alone blocked the flow-induced increase in O(2)(-) production (3.5 +/- 1.7 vs. 4.5 +/- 2.8 AU/s and 8.2 +/- 2.1 vs. 10.6 +/- 2.8 AU/s, respectively). Furosemide decreased flow-induced O(2)(-) production by 55% (37.3 +/- 5.2 to 16.8 +/- 2.8 AU/s; P < 0.002); however, dimethylamiloride had no effect. Finally, we examined whether changes in mechanical forces are involved in flow-induced O(2)(-) production by using a Na free solution to perfuse TALs. In the absence of NaCl, luminal flow enhanced O(2)(-) production (1.5 +/- 0.5 to 13.5 +/- 1.1 AU/s; P < 0.001), approximately 50% less stimulation than when flow was increased in the presence of luminal NaCl. We conclude that flow stimulates O(2)(-) production in TALs via activation of NADPH oxidase and that NaCl absorption due to Na-K-2Cl cotransport and flow associated mechanical factors contribute equally to this process. PMID- 17132869 TI - On experimental design and discourse in plasticity research. AB - Communication in the stem cell field requires a common understanding of terminology and that "plasticity" phenomena are model- and, perhaps, species dependent. Plasticity has generally been applied to unexpected differentiative events; will the term cease being useful when these unexpected pathways become recognized as normative? Four pathways of cell plasticity have now been recognized: (1) facultative, intraorgan self-renewing stem cells; (2) reversion of differentiated cells to blastema-like appearances, common in amphibians, perhaps restricted to neoplasia in mammals; (3) cells of one lineage directly changing to differentiation of another lineage cued by microenvironemental signals; (4) cell-cell fusion leading to changes in differentiation of the "incoming" cell in response to cytoplasmic and perhaps nuclear cues. In all of these, "differentiation" must be understood as a reflection of gene expression that is a highly intricate system of parallel, i.e., nonlinear molecular interactions. Present controversies regarding the plasticity of adult stem cells may be explained both by differences in experimental variables and techniques as well as by differing nonscientific, political, and/or polemical needs of investigators and commentators. Some of the variables in transplantation experiments, which are likely to be important in experimental outcome, but rarely addressed in interpretation of data, are the age of the cell donor and of the strain of mice or species used, the isolation technique used to obtain the putative stem cells, and the inherent effects of transgenic markers used to identify the donor or host cells. Also of great importance, but rarely controlled for in experimental design and interpretation, are the reproducibility and sensitivity of methods used to detect the markers of donor origin, the capacity of differentiated tissue to silence transgenes or alter marker expression, and- finally and most importantly--the different signals that influence plasticity phenomena in very different types of injury and regeneration. In different models of injury there are likely to be significant differences in promoting cell localization, proliferation, and predominance of "plasticity pathway," if any are involved, in determining outcome. PMID- 17132868 TI - Adult stem cell plasticity: introduction to the first issue of stem cell reviews. PMID- 17132870 TI - Bone marrow and brain: unexpected allies or accidental acquaintances? AB - Almost 10 yr have passed since we first entertained the idea that circulating blood cells originating in the bone marrow might contribute to central nervous system (CNS) cell lineages. Initially, we showed that microgalia are derived from bone marrow stem cells (BMSCs). Later, we found that BMSCs might also give rise to other neural cells--even neurons. We made our first observations in rodents, and then we examined human samples. In the brains of female patients who received bone marrow from male donors there are cells with neurons-specific markers together with the Y chromosome suggesting that they are the progeny donor bone marrow cells. All of our data and numerous recent experiments in the field suggest that circulating BM cells are quite plastic. Thus, they might be used in regenerative medicine to help heal or replace lost tissue in many different organs. PMID- 17132871 TI - Engraftment of bone marrow-derived epithelial cells. AB - The long-held concept that transplanted bone marrow (BM)-derived cells contribute only to cells of the hematopoietic system was challenged by data from our laboratory showing that a single male BM-derived cell could not only reconstitute the hematopoietic system of an irradiated female recipient, but could also lead to the generation of mature BM-derived epithelial cells in the liver, lung, skin, and gastrointestinal tract. Careful costaining and single-cell analyses have been used to rule out false positive cells due to inadequate detection techniques in microscopy or cell overlay. Since this initial discovery, we have sought to understand the mechanisms underlying the formation of BM-derived epithelial cells, and to evaluate their therapeutic use for gene therapy and/or tissue regeneration. Several reports have shown that donor BM-derived cells, possibly macrophages, can fuse with existing host epithelial cells to form heterokaryons that express both donor and tissue-specific markers. While this is certainly true for murine tyrosinemia models, we have used a Cre-lox system to demonstrate that fusion is not a requirement for the generation of BM-derived epithelial cells and is likely not responsible for the BM-derived epithelial cells generated after standard BM transplantation. In a proof of principal experiment for potential gene therapy applications, we have shown that autologous BM-derived cells transfected with a transgene prior to BM transplantation are able to develop into mature type-II pneumocytes that express the transgene. We also discuss future research directions in the field and the therapeutic potential of BM-derived epithelia, including ongoing work to test whether combined cell and gene therapy can be used therapeutically in preclinical mouse models of human disease. PMID- 17132872 TI - The stem cell continuum: considerations on the heterogeneity and plasticity of marrow stem cells. AB - Traditional models of hematopoiesis have been hierarchical. Recent evidence showing that marrow stem cells are a cycling population and that the hematopoietic phenotype of these cells reversibly changes with cycle transit have suggested a continuum model of stem cell regulators. Studies on marrow cell conversion to lung cells have extended this continuum to cycle-related differentiation into nonhematopoietic stem cells. We postulate that stem cells transiting cell cycle continually change their chromatin structure, thus providing different windows of transcriptional opportunity and a continually changing phenotype. Final outcomes with this continuum model would be determined by the specific chromatin state of the cell and the presence of specific differentiation inducers. PMID- 17132874 TI - Stem cell plasticity: a rare cell, not a rare event. AB - Purification to homogeneity for a rare stem cell (SC) population by both function and phenotype is a prerequisite to determine if SCs can change their fate (plasticity). Since cell fate determination has been suggested by both external environmental cues and intrinsic gene regulation, plasticity should be studied using both influences. Different frequencies of marrow SC plasticity may be attributed to either different isolation technologies or different developmental stage SCs with more or less multipotentiality. Tissue-specific SCs may reside in marrow, or alternatively, primitive marrow SC may respond directly to regenerative signals by migration to injury sites and repairing the damaged tissue. It is important to dissect the relationship between primitive/tissue specific SCs and regenerative signals. PMID- 17132873 TI - Stem cell plasticity: the debate begins to clarify. AB - The stem cell story begins with the recognition of the regenerative powers of the head of the Lernean Hydra and the human liver (Prometheus) by the ancient Greeks. In modern times, the adult human stem cell has been epitomized by the hematopoietic stem cell in the bone marrow. More recently, bone marrow derived cells were reported to contribute to nonhematopoietic organs, suggesting a level of plasticity not previously expected. However, other reports failed to repeat some of these results, resulting in a heated debate on the plasticity of adult stem cells that has crossed over into the public domain and become a matter of political impact on the use of embryonic vs adult stem cells for organ regeneration or gene therapy. This review discusses the current status of the "plasticity" debate and presents existing data on detection methodology, underlying mechanisms, physiological implications, and clinical significance. PMID- 17132875 TI - Multipotent adult progenitor cell and stem cell plasticity. AB - Stem cells are defined by their biological function. A stem cell is an undifferentiated cell that self-renews to maintain the stem cell pool and at the single-cell level differentiates into more than one mature, functional cell. In addition, when transplanted, a stem cell should be capable of replacing a damaged organ or tissue for the lifetime of the recipient. Some would argue that stem cells should also be capable of functionally integrating into nondamaged tissues. Stem cells are critical to both embryogenesis and postnatal life. PMID- 17132876 TI - Liver repair by intra- and extrahepatic progenitors. AB - Despite its remarkable capacity for endogenous regeneration, the mammalian liver is vulnerable to a number of chronic or acute conditions that exceed or circumvent the proliferative capabilities of its mature cell complement. Bipotential hepatic progenitors, or "oval cells," have been shown to contribute to organ regeneration under such circumstances, both in human patients and in animal models. These progenitors are attractive agents for cell therapy, but have thus far proven challenging to isolate and manipulate. New reports indicating that transplanted bone marrow cells (BMCs) can also generate hepatocytes and contribute to liver repair have attracted considerable attention, because these cells are familiar and accessible to both clinicians and scientists. Recently, the issue of whether nuclear transfer (via cell fusion between donor BMC and recipient hepatocyte) or previously unrecognized differentiation potential (i.e., plasticity/transdifferentiation of BMC) is the primary origin of donor-derived hepatocytes has generated considerable controversy. In the liver, most evidence supports cell fusion as the key agent in the reversal of hepatopathology. However, regardless of their origin, the frequency of hepatocyte correction events is low. As is the case for the delivery of intrahepatic progenitors, substantial improvements in the understanding of this process will be needed before clinical application becomes practical. PMID- 17132877 TI - Plasticity and tissue regenerative potential of bone marrow-derived cells. AB - Diverse in vivo studies have suggested that adult stem cells might have the ability to differentiate into cell types other than those of the tissues in which they reside or derive during embryonic development. This idea of stem cell "plasticity" has led investigators to hypothesize that, similar to embryonic stem cells, adult stem cells might have unlimited tissue regenerative potential in vivo, and therefore, broad and novel therapeutic applications. Since the beginning of these observations, our group has critically examined these exciting possibilities for mouse bone marrow-derived cells by taking advantage of well characterized models of tissue regeneration, Cre/lox technology, and novel stem cell isolation protocols. Our experimental evidence does not support plasticity of hematopoietic stem cells as a frequent physiological event, but rather indicates that cell fusion could account for reported cases of hematopoietic stem cell plasticity or "transdifferentiation" in vivo. Our studies highlight the need for meticulous technical controls during the isolation, transplantation, tracking, and analysis of bone marrow-derived cells during in vivo studies on plasticity. Further studies will be necessary to better define experimental conditions and criteria to unequivocally prove or reject plasticity in vivo. In this review, we focus on results from several studies from our laboratory, and discuss their conclusions and implications. PMID- 17132878 TI - Plasticity of epidermal stem cells: survival in various environments. AB - The keratinocyte cell compartment in the continuously renewing epidermis of the skin is maintained by undifferentiated, self-renewing stem cells. We show that a small subpopulation of epidermal stem cells (EpiSCs) have the capacity to integrate into multiple tissues. These EpiSCs can change their phenotype in direct response of changes in cytokines in vitro, changes in cocultured cells, after injection into damaged environments in vivo. These changes appear to be unrelated to the age of the EpiSC. Even though we can isolate these cells and show that the age of thses cells appears to be irrelevant to this multipotent function, we still do not know how such cells are defined within a tissue or what the life span of a multipotent stem cell is. PMID- 17132879 TI - Fasting insulin, adiponectin, hs-CRP levels, and the components of metabolic syndrome. AB - AIM: in this study we report the relationship between fasting plasma insulin, C reactive protein, adiponectin levels and the components of metabolic syndrome. METHODS: for the diagnosis of metabolic syndrome we used the modified NCEP ATP criteria for Asian people. Complete routine physical examinations were performed to all subjects including blood pressure, waist circumference. After 12-hours fasting, blood sample was taken for fasting plasma glucose, fasting insulin, and lipid profiles. Fasting insulin was determined by RIA, adiponectin by ELISA, and hs-CRP by sensitive immunometric assay. According to NCEP ATP III, the metabolic syndrome consists of five components. Subjects fulfilling the studied criteria were divided into five groups according to the component/components they had. RESULTS: during the study, 118 adult individuals can be covered, including 88 patients (68,8%) with metabolic syndrome. They consist of 19 subjects who had only 1 component, 21 subjects had 2 components, 31 subjects with 3 components, 34 subjects with 4 components, and 23 subjects had 5 component. All subjects were overweight/obese (BMI > 23 kg/m), but patients with metabolic syndrome were significantly more obese compared to the non-metabolic syndrome (p < 0.05). There was no significant difference in age among all individuals. Fasting insulin, hs CRP levels is increasing with the increasing number of components of metabolic syndrome, being higher among those with 3-5 components, while adiponectin is decreasing with the number of the components. Fasting insulin 3,5+1,1 uU/ml, 3,6+1,4 uU/ml, 5,9+1,8 uU/ml, 7,8+2,1 uU/ml, 7,9+2,3 uU/ml respectively, hs-CRP 2,8+1,2 mg/L, 5.6+3.4 mg/L, 7.4+4.4 mg/L, 9.0+4.7 mg/L, 9.5+3,9 mg/L, and adiponectin 9.1+3.5 ng/ml, 8.6+1.6 ng/ml, 3.4+1.2 ng/ml, 3.2+1.3 ng/ml, 2.8+0.9 ng/ml in 3,4,5 components respectively. CONCLUSION: there is a relationship between the number of components of metabolic syndrome and the increasing levels of fasting insulin, and hs-CRP, and low levels of adiponectin. These may explain in part the risk of cardiovascular events among individuals with metabolic syndrome. PMID- 17132880 TI - The effect of aerobic exercise on blood and plasma viscosity on cardiac health club participants. AB - AIM: to analyze the effect of SJS aerobic exercise on blood and plasma viscosity. METHODS: the study was performed on 30 subjects with an age span of 40 to 60 years. Subjects participated in SJS aerobic exercise of moderate intensity of 40 to 45 minutes duration, three times a week for 9 to 12 weeks. Five milliliters of blood were collected into K3EDTA container to assess blood and plasma viscosity prior to the program and following the completion of the SJS program. Blood and plasma viscosity was measured using Brookfield LVDV-III viscometer using rotational method principle. RESULTS: this study demonstrated a significant decrease in blood viscosity (2.94%, p = 0.03) and insignificant decrease in plasma viscosity in subjects following SJS aerobic exercise compared to prior exercise. CONCLUSION: this study proved that SJS aerobic exercise of moderate intensity of 40 to 45 minutes duration times a week for 9 to 12 weeks gave the benefit of lowering blood viscosity, which contributes to reducing the risk of coronary heart disease. PMID- 17132881 TI - Estimating the annual cost of overactive bladder in Indonesia. AB - AIM: to estimate the annual economic cost and quality of life related to OAB in Indonesia population by taking into account the direct cost, value of lost productivity and cost consequences associated with OAB. METHODS: cross sectional study was done in Geriatric Clinic and Urogynaecology Clinic in Dr. Cipto Mangunkusumo Hospital Jakarta from July 2005 to March 2006. Primary outcome of this study was annual cost of OAB, symptom and quality of life of OAB patients. Cost data related to personal routine care of OAB and transportation expenditures were obtained by using questionnaires modified from Dowell Bryant incontinence cost index validated questioner. Data related to quality of life were obtained by OAB-q questioner. Subjects included 30 male and female OAB patients aged 18-100 years. RESULTS: most of patients were female (96.7%). Median of age was 62.5 (30 93) years old, 56.7% patients were elderly (age more than 60 years). Median of total annual cost of OAB was Rp. 2,850,000,-. Median of total personal cost which consist of routine personal care costs and treatment costs were Rp. 2,850,000. Median of total cost which is expended by government for routine personal and treatment of OAB was Rp.2,500,000,- . Median score of symptom severity was 62.5. Quality of life score was divided into coping, concern, sleep, and social item. Median of coping score was 50.0, concern score was 43.1, sleep score was 50.8, and social score was 38.8. Median of total quality of life score was 44.7. Maximum score of symptom severity and quality of life should be 98.8. The higher the score, the more severe the symptom, but the better is the quality of life. CONCLUSION: total annual cost of OAB was Rp. 2,850,000, Quality of life of OAB patient was somewhat poor. PMID- 17132882 TI - The correlation between free thyroxine levels and left ventricular mass in Graves' disease. AB - AIM: to determine the correlation between free thyroxine level and left ventricular mass in newly diagnoses Graves' disease. METHODS: seventeen patients with newly diagnosed Graves' disease were studied. Inclusion criteria was new case of Graves' disease, no previous history or clinical evidence of coronary artery disease, cardiac valve disease, diabetes mellitus, chronic kidney disease, not taking antithyroid drugs and any drugs that could affect the heart (beta blockers, ACE inhibitor). Echocardiographic indices of left ventricular mass was obtained. RESULTS: there were no correlation between free thyroxine levels and left ventricular mass, systolic and diastolic blood pressure. There was a correlation between pulse rate and left ventricular mass in Graves' disease. CONCLUSION: there was no correlation between free thyroxine level and left ventricular mass in Graves' disease. PMID- 17132883 TI - Lipid profiles of acute coronary syndrome patients hospitalized in ICCU of Cipto Mangunkusumo Hospital. AB - AIM: to see which component of the triad lipid that has more important role and frequently found in patients with acute coronary syndrome, focusing on HDL cholesterol. METHODS: a lipid profile study has been conducted in 391 patients with acute coronary syndrome (ACS), who have been hospitalized in ICCU of Cipto Mangunkusumo Hospital since January 1st, 2001 - December 31st, 2005. RESULTS: there were 294 male patients (75.2%) and 97 female patients (24.8%), from 25-89 years age group, with mean value 57.35 +/- 11.05. The mean value of total cholesterol level was 205.23 mg/dl +/- 54.84. The LDL cholesterol level was 136.16 mg/dl +/- 47.29. The mean HDL cholesterol level was 42.84 mg/dl +/- 10.28, mean triglycerides level was 157.25 mg/dl +/- 100.16. There were 82 patients (21%) with high total cholesterol (> 240 mg/dl), 102 patients (26.1%) with high and very high LDL cholesterol level (> 160 mg/dl), 152 (38.6%) patients with low HDL cholesterol level (< 40 mg/dl), consisted of 126 male patients or 42.9% of total male patients, 26 female patients or 26.8% of total female patients. The number of patients with high / very high triglycerides level (> 200 mg/dl or 500 mg/dl) was 84 patients (21.5%). The number of patients with optimal lipid level, total cholesterol level < 200 mg/dl was 191 patients (48.8%); 82 patients (21%) had LDL cholesterol level < 100 mg/dl, 23 patients (5.9%) had HDL cholesterol level > 60 mg/dl, 226 patients (57.8%) had triglycerides level < 150 mg/dl. There were 260 patients with atherogenic lipid profile (64%), 135 patients with borderline lipid profile (34.5%), and optimal lipid profile was found only in 6 patients (1,5%). The mean value of HDL cholesterol level in male patients was 41.75 mg/dl +/- 9.9, while HDL cholesterol level in female patients was 46.16 mg/dl +/- 10.74. Principally, lipid profiles of all age groups were evenly distributed. ACS patients with history of family heart disease had a more atherogenic lipid profile compared to lipid profile of patients with hypertension, diabetes mellitus, and cigarette smoking. CONCLUSION: atherogenic lipid profile is more common risk factor in ACS patients than in other non-lipid risk factors. This study also demonstrated that the atherogenic lipid is most commonly found in patient with low HDL cholesterol level and it is least frequently found in patient with optimal lipid profile. PMID- 17132884 TI - The prevalence of hypokalemia in hospitalized patients with infectious diseases problem at Cipto Mangunkusumo Hospital, Jakarta. AB - AIM: to know the prevalence of hypokalemia that occurs in hospitalized patients with infectious diseases. METHODS: a cross sectional study was carried out in the internal ward Cipto Mangunkusumo General Hospital in Jakarta from December 2005 until June 2006. All hospitalized patients with infectious diseases receiving "replacement solution" were included in this study. We collected the blood sample to perform the serum potassium level at the time of admission and discharge. RESULTS: one hundred and five patients were enrolled in this study; consisting of 44 males and 61 females. The age ranged from 14 to 70 years old. The most common infectious diseases were dengue fever, while the underlying diseases were hepatobiliary disorders. "Replacement solutions" which were given, were ringer's lactate 91%, normal saline 8%, and ringer's acetate 1%. Prevalence of hypokalemia among the hospitalized patients, on admission was 24 patients (23%) and during hospitalization was 39 patients (37%). The mean level of hypokalemia on admission was 3.11 + SD 0.37 mEq/L (range from 1.7 to 3.4 mEq/L) and during hospitalization was 3.13 + SD 0.25 mEq/L (range from 2.5 to 3.4 mEq/L). On admission, the ratio of mild : moderate : severe hypokalemia is 22 : 2 : 1. And at discharge the ratio of mild to moderate hypokalemia becomes 19 : 6. CONCLUSION: the prevalence of hypokalemia in hospitalized patients with infectious disease in Cipto Mangkunkusumo's Hospital, Jakarta is 23%. Further studies are needed to know the contributing factors including the usage of 'intravenous fluid' in relations to hypokalemic state during hospitalization. PMID- 17132885 TI - A study on immunopathogenetic mechanisms of atherosclerotic process caused by chronic infection of Chlamydia pneumoniae in rats (Ratus novergicus). AB - AIM: this study was aimed to determine the correlations between duration of Chlamydia pneumoniae infection and the development of atherosclerotic process in white-rats' (Ratus novergicus) aorta. METHODS: this is an experimental study which examined the expression of TNFa, IL-1b, IL-8, adhesion molecule of VCAM-1 and the development of foam cells associated with atherosclerotic process in white-rats' aorta. There were 32 male rats, +/- 6 weeks of age, divided into 4 groups: control group (K) without infection, and 3 groups with infection through nasal and oral inoculation of Chlamydia pneumoniae by single dose of 5 x 105 in amount of 35 microl. The first group (P1) was preserved for 5(1/2) months period, the second group (P2) was preserved for 7(1/2) months and the third group (P3) was preserved for 9(1/2) months. At the end of study, histological slides were made from aortic tissues in order to study the development of atherosclerotic process by examining foam cells and cytokines expression of TNFa, IL-1b, IL-8 and VCAM-1. Foam cells examination was performed by Hematoxcillin-eosin staining, while indirect immunohistochemistry staining was used to examine the expression of TNFa, IL-1b, IL-8 and VCAM-1. Afterward, the amount of foam cells and cytokines expression was measured. The study result was analyzed by ANOVA. RESULTS: there was increased expression of TNFa, IL-1b, IL-8, VCAM-1and increased foam cells formation (extended atherosclerosis area) in the aortic tissues infected by Chlamydia pneumoniae (5(1/2) months, 7(1/2) months and 9(1/2) months), which was significantly different compared to the control group. The result of ANOVA revealed that the most important factor in tissue injury is foam cells development induced by VCAM-1 and IL-8 in all of phases (characterized by most abundant neutrophil infiltration). It indicated the infection caused by extracellular pathogenic agent, which established the fatty streak (acute phase) in 5(1/2) months period. In the group with 7(1/2) months infection period, TNFa also had important roles (characterized by increased monocytes and lymphocytes infiltration), indicating that there was negative-gram pathogenic agent with intracellular infection, which caused a progressive atherosclerotic process, and development of fibrosis / atherosclerotic plaque (sub acute phase). In 9(1/2) months infection period, there was large thrombus containing a lot of leukocytes in the aorta (chronic phase). CONCLUSION: based on the result of this study, it may be concluded that Chlamydia pneumoniae may cause atherosclerotic process in aorta. Extracellular infection of Chlamydia pneumoniae occurs in all of phases and intracellular infection begins in sub-acute phase. On 5(1/2) months period, fatty streak is developed (acute phase); on 7(1/2) months period, there is atherosclerotic plaque (sub acute phase); and on 9(1/2) period, there is large thrombus containing a lot of leukocytes (a progressive chronic phase). PMID- 17132886 TI - Scleroderma in a young man. AB - Scleroderma is a chronic disorder, slowly progressing characterized by diffuse fibrosis of the skin and internal organs. There are functional and structural abnormalities small blood vessels, fibrosis of the skin and internal organs, immune system activation and autoimmunity. Female predominance is most pronounced during mid and late childbearing age, women to men ratio is 7-12 : 1. In this case, we found a diffuse cutaneus scleroderma. Skin biopsy result showed hyperkeratosis, increased dermis thickness with deposition of collagen. The antigenic specificities of antibodies were related to distinct connective tissue disease. The diffuse cutaneus subset was associated with antibodies to topoisomerase I, formerly described as Scl-70 or Scl-86. Anti Scl-70 antibodies were shown in many studies as a marker antibodies for SSC. Colchicine was given in this case, which has the effects of microtubule assembly inhibition and interference with transcellular movement of collagen. During colchicine treatment serum hydroxyproline increased and serum proline decreased, suggesting increased collagen degradation. PMID- 17132887 TI - Hypertension: diagnostic problem, challenge and dilemmas. AB - An elevated arterial pressure is probably the most important public health problem. The prevalence of hypertension depends on both the racial composition of the population studied and the criteria used to define the condition. Patients with hypertension die prematurely, the most common cause of death is heart disease, stroke, and renal failure. The JNC-7 report has introduced a new classification that includes term "pre-hypertension". The new classification may make a new dilemma in the management since the main treatment of pre-hypertension is the lifestyle changes. What do we recommend to lean patients with pre hypertension who are already following a prudent lifestyle? The ultimate public health goal of antihypertensive therapy is the reduction of cardiovascular and renal morbidity and mortality. The question is, what should the blood pressure goal be? For patients without any compiling condition, it was assumed that 140/90 mmHg was desired treatment target level. For diabetic patient, it is reasonable to target a blood pressure well within the normal range, at most 130/80 mmHg. Some newer studies suggest that the target of treatment may force the recommended goal even lower, even for the patient without any compiling condition. For elderly individuals, a goal of 140/90 mmHg is appropriate. Hypertension during pregnancy is one of important aspects, since blood pressure during pregnancy may change related to the gestational age. This is always subject of discussion because there is no evidence that pharmacologic treatment results in improved neonatal outcomes, lower blood pressure may, in fact uteroplacental perfusion and thereby jeopardize fetal development. It means that more attention is needed for hypertensive patients not only for treatment regimens choice but also the blood pressure target. The accurate measurement of blood pressure is the sine qua non for successful management of hypertension. The operator should be trained and regularly retrained in the standardized technique, to avoid the chance of mismanagement. PMID- 17132888 TI - Severe ankylosing spondylitis. PMID- 17132889 TI - The competency of internists in holistic global care to support healthy Indonesia 2010. AB - All patients are entitled to good standards of practice and care from their doctors. The improved health of all peoples is the main goal of medical education, including for the education of internal medicine registrars. The future development of the direction and goal of education, the level of competence of internal medicine specialists, and the process of learning will be related to preparing the internal medicine specialist to have global competitive advantage. Identification of general competencies is the first step in a long term effort designed to emphasize educational outcome, for assessment in residency programs, and in the accreditation process. To achieve that competence, a variety of learning opportunities need to be provided in order that the resident can achieve the necessary knowledge, skills, attitude, and behaviors. Identification of the role and function of internal medicine specialists is needed prior to the development of the general competencies. As educational objectives, the competencies fall into two main categories: knowledge-based and performance-based. Knowledge-based competency has two components, medical knowledge (bio-science and clinical medicine) and contextual knowledge (epidemiology, health service organization, and human behavior). The performance base has two components, intellectual skills and the interpersonal skills. Besides the two main categories of educational objectives, there are behavioral objectives that residents must achieve through the educational program, to ensure that residents are able to deal with a range of prescribed clinical situations effectively, safely, humanely, and economically. The achievement of behavioral objectives will ensure, at least in part, that the doctor will implement good medical practice. The index clinical/community situations (ICS) on which the educational objectives will be based are taken from diseases and illnesses that occur in clinical and community settings. No resident can master all medicine there is to know, as there are no limits to what can be known about medicine. It is important to make choices in selecting what residents should learn by analyzing the ICS. PMID- 17132890 TI - Update in the management of obesity. AB - Significant increase of obesity prevalence in almost all countries in the world recently has had obesity as a global health problem, and WHO in 1998 defined it as "the global epidemic". Simply, obesity is defined as an excessive fat accumulation in fat tissue due to imbalance of energy intake and expenditure. Body mass index is a simple method for defining the degree of overweight and obesity, however, waist circumference is the preferred measure of abdominal obesity because it has greater relationship with the risk of metabolic and cardiovascular diseases. Body fatness reflects the interactions of development, environment and genetic factors. The role of genetic factors has already existed, nevertheless, environment factors are likely more important in developing obesity. Increased mortality among the obese is evident for several life threatening diseases including type 2 diabetes, cardiovascular disease, gallbladder disease, and hormone-sensitive and gastrointestinal cancers. Risks are also higher for some non-fatal conditions such as back pain, arthritis, infertility and, in many westernized countries, poor psychosocial functioning. Obesity is not only threatening health, also impacts on high economic and social cost. Effective prevention of obesity should be focused to high risk individuals or groups. Individuals who have some existing weight-related problems and those with a high risk of developing obesity co-morbidity such as cardiovascular disease and type 2 diabetes should be a key priority in this prevention strategy. Although weight loss in obese persons of any age can improve obesity-related medical complications, physical function, and quality of life, the primary purpose for weigh-loss therapy may differ across age group. The current therapeutic tools available for weight management are: (1) lifestyle intervention involving diet, physical activity, and behavior modification; (2) pharmacotherapy; and (3) surgery. Moderate weight loss (5-10% of initial weight) by any programs is a realistic target in management of obesity associated with improvement of risk factors of metabolic and cardiovascular diseases. PMID- 17132891 TI - Malnutrition-inflammation syndrome in a hemodialysis population: the influence of polymorphic IL-6-174 and IL-10-1082 genes. AB - AIM: To find out inflammation prevalence in hemodialysis patients and whether polymorphic gene IL-6-174 and IL-10-1082 had prominent factors in malnutrition inflammation syndrome. METHODS: A study on 81 patients who were on hemodialysis twice a week, 5 hours each session has been conducted. The subjects had no other co-morbidities and all of them used reprocessed diacetate cellulose dialyzers. RESULTS: It was obtained that CRP blood level (6.23+/- 5.57 mg/L), inflammation prevalence (23.5%), and malnutrition inflammation score (6.7) were lower compared with the data from Europe and the United States. Out of 64 patients examined, IL 6-174GG 95.31%, CC 3.13% and GC 1.56%, IL-1082AA 89.06%, GA 10.94%, but absence of GG genotype. Considering the scanty amount of allele C in IL-6-174 gene and G allele in IL-10-1082 gene, based on the statistic analysis performed, it did not reveal the influence of the difference in allele on the clinical manifestation. The proportion of these alleles were almost similar to that obtained in Korea, Japan and China, but it was different from that obtained in the US and in Europe. A very resolute impression was obtained in HD patients in Jakarta that IL-6-174GG gene was protective in nature whereas IL-10-1082 AA gene had a less considerable role. CONCLUSION: The prevalence of inflammation and malnutrition-inflammation parameters (CRP, malnutrition Inflammation Score) in our HD patients were lower than that reported in Western countries. This might be related to the low prevalence of IL-6-174C allele in our population. PMID- 17132892 TI - Unintended consequences of plant transformation: a molecular insight. AB - Plant genomes are dynamic structures having both the system to maintain and accurately reproduce the information encoded therein and the ability to accept more or less random changes, which is one of the foundations of evolution. Crop improvement and various uncontrolled stress factors can induce unintended genetic and epigenetic variations. In this review it is attempted to summarize factors causing such changes and the molecular nature of these variations in transgenic plants. Unintended effects in transgenic plants can be divided into three main groups: first, pleiotropic effects of integrated DNA on the host plant genome; second, the influence of the integration site and transgene architecture on transgene expression level and stability; and third, the effect of various stresses related to tissue handling, regeneration and clonal propagation. Many of these factors are recently being redefined due to new researches, which apply modern highly sensitive analytical techniques and sequenced model organisms. The ability to inspect large portions of genomes clearly shows that tissue culture contributes to a vast majority of observed genetic and epigenetic changes. Nevertheless, monitoring of thousands transcripts, proteins and metabolites reveals that unintended variation most often falls in the range of natural differences between landraces or varieties. We expect that an increasing amount of evidence on many important crop species will support these observations in the nearest future. PMID- 17132893 TI - Analysis of the wheat endosperm transcriptome. AB - Among the cereals, wheat is the most widely grown geographically and is part of the staple diet in much of the world. Understanding how the cereal endosperm develops and functions will help generate better tools to manipulate grain qualities important to end-users. We used a genomics approach to identify and characterize genes that are expressed in the wheat endosperm. We analyzed the 17,949 publicly available wheat endosperm EST sequences to identify genes involved in the biological processes that occur within this tissue. Clustering and assembly of the ESTs resulted in the identification of 6,187 tentative unique genes, 2,358 of which formed contigs and 3,829 remained as singletons. A BLAST similarity search against the NCBI non-redundant sequence database revealed abundant messages for storage proteins, putative defense proteins, and proteins involved in starch and sucrose metabolism. The level of abundance of the putatively identified genes reflects the physiology of the developing endosperm. Half of the identified genes have unknown functions. Approximately 61% of the endosperm ESTs has been tentatively mapped in the hexaploid wheat genome. Using microarrays for global RNA profiling, we identified endosperm genes that are specifically up regulated in the developing grain. PMID- 17132894 TI - Development of loop-mediated isothermal amplification (LAMP)-based SNP markers for shelf-life in melon (Cucumis melo L.). AB - In this study, LAMP markers linked to shelf-life in melon (Cucumis melo L.) were developed by converting a cleaved amplified polymorphic sequences (CAPS) marker (C2). The CAPS-PCR fragments from the long-shelf-life melon (O-3) and short-shelf life melon (Nat-2) were cloned and sequenced to construct LAMP primers. A single nucleotide polymorphism (SNP) was identified between O-3 and Nat-2. LAMP primers were designed to detect the SNP. In the LAMP reaction to detect long-shelf-life melon, the turbidity of the templates using O-3, F1, homozygous long-shelf-life F2 lines and heterozygous long-shelf-life F2 lines started to increase after 40 min. In contrast, the turbidity of Nat-2 and homozygous short-shelf-life F2 lines did not increase even after 90 min. In the LAMP reaction to detect short-shelf life melon, the turbidity of the templates using Nat-2, F1, homozygous short shelf-life F2 lines and heterozygous long-shelf-life F2 lines started to increase after 40 min. But the turbidity of O-3 and homozygous long-shelf-life F2 lines did not increase after 90 min. This attests to the high reliability and usefulness of LAMP for marker-assisted selection. PMID- 17132895 TI - Agrobacterium-mediated transformation of yellow lupin to generate callus tissue producing HBV surface antigen in a long-term culture. AB - The idea of an oral vaccine administered as a portion of plant tissue requires a high level of antigen production. An improved protocol for the induction of transgenic yellow lupin calli or tumours, reaching 44% of transformation rate, is presented here. It has been developed by using the nptII marker gene and the uidA reporter gene as well as various Agrobacterium strains and plant explants. This method of seedling and hypocotyl transformation was applied to raise calli or tumours producing a small surface antigen of Hepatitis B Virus (S-HBsAg). Lupin tissue lines were long-term cultured on selection media maintaining the growth rate and high expression level of the native form of S-HBs, up to 6 microg per g of fresh tissue. PMID- 17132896 TI - Effects of interaction between pollen coat eluates and pistil at the molecular level in self-compatible and self-incompatible plants of Lolium multiflorum Lam. AB - Two-dimensional electrophoresis (2-DE) of soluble proteins and enzymes was performed and specific activities of 5 enzymes (esterase, pectinesterase, acid phosphatase, protease and diaphorase) were determined in stigmas of Lolium multiflorum (Italian ryegrass) treated with self or foreign pollen coat eluates (pc). Also, a low-molecular-weight fraction of the treated self-compatible (SC) and self-incompatible (SI) stigmas was analyzed by high-pressure liquid chromatography (HPLC). The treatment of stigmas with foreign pollen induced the loss of 42% of the control sample proteins in SC plants but only of 5.5% in SI plants. In contrast, the treatment of stigmas with foreign pollen induced the loss of 15% proteins in SC plants and of 29% in SI plants. Specific activities of esterase, pectinesterase and diaphorase were higher in SC than in SI stigmas. The 2-DE enzyme patterns indicated qualitative relationships between the presence of some isoforms of acid phosphatase or protease and the treatment with self or foreign pc in SC and SI stigmas. No changes were observed in HPLC profiles of the low-molecular-weight fraction from SC and SI stigmas treated or not with pc. The presented results revealed different reactions of SC and SI stigmas to the treatment with self or foreign pc. Further investigations may explain if any of the observed reactions represent specific reorientations in the style, facilitating cross- or self-pollination. PMID- 17132897 TI - Xenia effect on seed and embryo size in cotton (Gossypium hirsutum L.). AB - The term xenia was coined to describe the effect of foreign pollen on the development and characters of the seed. To study its importance and consequences for various seed traits in cotton (Gossypium hirsutum L.), the effect of pollen genotype on seed and embryo weight was studied with seeds from 15 F1 hybrids. Cross-fertilization changed seed weight by up to 7.0% in relation to self fertilization. Xenia effect significantly increased embryo weight of cross fertilized seeds, by up to 14.4% in comparison to self-fertilized seeds. Seeds of some crosses had a lower hull content than corresponding selfed seeds. On average, the xenia effect was greater for embryo weight than for seed weight. However, in some crosses there was no difference between cross- and self fertilized seeds for seed weight, embryo weight, moisture content and hull content. Positive xenia effects for seed weight and embryo weight may help us to establish uniform stands of vigorous hybrid seedlings, especially under unfavourable conditions. Also, larger seed and embryo weight, along with lower hull content, could result in higher oil yield. Therefore, careful choosing of genotypes as parents and of cross direction in the production of hybrid seed is very important in cotton. PMID- 17132898 TI - Investigation of major gene for milk yield, milking speed, dry matter intake, and body weight in dairy cattle. AB - The main aim of this study was to determine if there exist any major gene for milk yield (MY), milking speed (MS), dry matter intake (DMI), and body weight (BW) recorded at various stages of lactation in first-lactation dairy cows (2543 observations from 320 cows) kept at the research farm of the Swiss Federal Institute of Technology between April 1994 and April 2004. Data were modelled based a simple repeatability covariance structure and analysed by using Bayesian segregation analyses. Gibbs sampling was used to make statistical inferences on posterior distributions; inferences were based on a single run of the Markov chain for each trait with 500,000 samples, with each 10th sample collected because of the high correlation among the samples. The posterior mean (+/-SD) of major gene variance was 2.61 (+/-2.46) for MY, 0.83 (+/-1.26) for MS, 4.37 (+/ 2.34) for DMI, and 2056.43 (+/-665.67) for BW. Highest posterior density regions for 3 of the 4 traits did not include 0 (except MS), which supported the evidence for major gene. With additional tests for agreement with Mendelian transmission probabilities, we could only confirm the existence of a major gene for MY, but not for MS, DMI, and BW. Expected Mendelian transmission probabilities and their model fits were also compared. PMID- 17132899 TI - Relative transcript abundance of oxytocin receptor gene in porcine uterus during luteolysis and early pregnancy. AB - The aim of the present study was to investigate transcript localization of the oxytocin receptor (OTR) gene in different cells of the porcine uterus during luteolysis and early pregnancy (days 14-16) using in situ hybridization (ISH). OTR mRNA was localized in the uterine luminal epithelium (LEC), glandular epithelium (GEC), stromal cells (SC) of the endometrium, in the longitudinal muscle layer (LM) and circular muscle layer (CM) of the myometrium. The OTR transcript was quantified by optical density units of silver grains. The OTR transcript levels in the endometrium and myometrium were statistically higher during luteolysis than during early pregnancy (P<0.05). Besides, during luteolysis, the mRNA level was higher in the LEC, GEC of the endometrium and LM of the myometrium compared to that observed in the SC of endometrium and CM of the myometrium, respectively (P<0.05). In summary: 1) the level of OTR mRNA in uterine tissues is higher during luteolysis compared to early pregnancy, 2) the OTR transcript level in endometrial cells did not correspond to the sensitivity of these cells to oxytocin (OT), 3) the myometrial expression of the OTR gene is appropriate to control contractile activity and secretion of PG during luteolysis. PMID- 17132900 TI - Genetic structure and phylogenetic relationships of the Polish Heavy horse. AB - In this study a wide range of genetic markers (12 microsatellites, 7 blood-group loci, 10 blood-protein loci) and mitochondrial DNA (mtDNA) were used to assess genetic diversity in Polish Heavy horses. Three random samples were sequenced for 421 bp of the mitochondrial D-loop region, but no clear phylogenetic patterns were seen in mtDNA variation. Both heterozygosity and diversity levels are fairly high in Polish Heavy horses. In phylogenetic analysis the draught horses form a distinct cluster that pairs with the true pony breeds. Within this 'cold-blooded' group, the Polish Heavy Horse clusters most closely with the Posavina breed from Croatia and the Breton breed from France. From the standpoint of genetic conservation, the Polish Heavy Horse does not appear to be in jeopardy. PMID- 17132901 TI - The retinal fascin gene 2 (FSCN2)--partial structural analysis and polymorphism detection in dogs with progressive retinal atrophy (PRA). AB - The retinal fascin 2 gene (FSCN2) underwent a molecular analysis, a search for polymorphisms and an evaluation as a candidate gene for retinopathies in dogs. Specific fragments of the gene encompassing partial exon 1 and intron 1, and exons 2-5 with respective introns were sequenced and these data were deposited in the GenBank database. Three distinct polymorphic sites detectable with PCR-RFLP were found--AM050719: g.237G>A, AM050719: g.525A>G, and AM050720: g.1071A>G. No positive associations between these polymorphisms and the PRA-clinical status were observed in the investigated population consisting of Poodles, American Cocker Spaniels, and English Cocker Spaniels. In spite of that, the FSCN2 gene remains an excellent candidate gene for retinopathies in dogs and the results can contribute to further research in this field. PMID- 17132902 TI - Epigenetic diagnostics of cancer--the application of DNA methylation markers. AB - In recent years it has become apparent that epigenetic events are potentially equally responsible for cancer initiation and progression as genetic abnormalities. DNA methylation is the main epigenetic modification in humans. Two DNA methylation lesions coexist in human neoplasms: hypermethylation of promoter regions of specific genes within a context of genomic hypomethylation. Aberrant methylation is found at early stages of carcinogenesis and distinct types of cancer exhibit specific patterns of methylation changes. Tumor specific DNA is readily obtainable from different clinical samples and methylation status analysis often permits sensitive disease detection. Methylation markers may also serve for prognostic and predictive purposes as they often reflect the metastatic potential and sensitivity to therapy. As current findings show a great potential of recently characterised methylation markers, more studies in the field are needed in the future. Large clinical studies of newly developed markers are especially needed. The review describes the diagnostic potential of DNA methylation markers. PMID- 17132903 TI - DNA damage in mouse lymphocytes exposed to curcumin and copper. AB - Dietary polyphenolics, such as curcumin, have shown antioxidant and anti inflammatory effects. Some antioxidants cause DNA strand breaks in excess of transition metal ions, such as copper. The aim of this study was to evaluate the in vitro effect of curcumin in the presence of increasing concentrations of copper to induce DNA damage in murine leukocytes by the comet assay. Balb-C mouse lymphocytes were exposed to 50 microM curcumin and various concentrations of copper (10 microM, 100 microM and 200 microM). Cellular DNA damage was detected by means of the alkaline comet assay. Our results show that 50 microM curcumin in the presence of 100-200 microM copper induced DNA damage in murine lymphocytes. Curcumin did not inhibit the oxidative DNA damage caused by 50 microM H2O2 in mouse lymphocytes. Moreover, 50 microM curcumin alone was capable of inducing DNA strand breaks under the tested conditions. The increased DNA damage by 50 mM curcumin was observed in the presence of various concentrations of copper, as detected by the alkaline comet assay. PMID- 17132904 TI - DNA microsatellite analysis in families with autosomal dominant polycystic kidney disease (ADPKD): the first Polish study. AB - BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common inherited renal disorders with genetic heterogeneity. Mutations of two known genes are responsible for this disease: PKD1 at 16p13.3 and PKD2 at 4q21-23. A majority of cases (85%) are caused by mutations in PKD1. Because direct mutation screening remains complex, we describe here the application of an efficient approach to studies based on highly informative dinucleotide and tetranucleotide repeats flanking genes PKD1 and PKD2. METHODS: For this study a series of microsatellites closely linked to locus PKD1 (D16S291, D16S663, D16S665, D16S283, D16S407, D16S475) and to locus PKD2 (D4S1563, D4S2929, D4S414, D4S1534, D4S423) were selected. Short (81-242 bp) DNA fragments containing the tandem repeats were amplified by polymerase chain reaction (PCR). The number of repeat units of microsatelite markers was determined by fluorescent capillary electrophoresis. RESULTS: DNA microsatellite analysis was performed in 25 Polish ADPKD families and established the type of disease (21 families PKD1-type, 1 family PKD2-type). CONCLUSIONS: While a disease-causing mutation in the PKD1 and PKD2 genes cannot be identified, DNA microsatellite analysis provided an early diagnosis and may be considered in ADPKD families. PMID- 17132905 TI - Genotyping of alpha-thalassemia in microcytic hypochromic anemia patients from North India. AB - Microcytic hypochromic anemia is a common condition in clinical practice and alpha-thalassemia has to be considered as a differential diagnosis. Molecular diagnosis of alpha-thalassemia is possible by polymerase chain reaction. The aim of this study was to evaluate the frequency of alpha-gene numbers in subjects with microcytosis. In total, 276 subjects with microcytic hypochromic anemia [MCV<80fl; MCH<27pg] were studied. These include 125 with thalassemia trait, 48 with thalassemia major, 26 with sickle-cell thalassemia, 15 with E beta thalassemia, 40 with iron-deficiency anemia, 8 with another hemolytic anemia, and 14 patients with no definite diagnosis. Genotyping for -alpha3.7 deletion, alpha4.2 deletion, Hb Constant Spring, and a-triplications was done with polymerase chain reaction. The overall frequency of -alpha3.7 deletion in 276 individuals is 12.7%. The calculated allele frequency for a-thalassemia is 0.09. The subgroup analysis showed that co-inheritance of a-deletion is more frequent with the sickle-cell mutation than in other groups. We were able to diagnose 1/3 of unexplained cases of microcytosis as a-thalassemia carriers. The a-gene mutation is quite common in the Indian subcontinent. Molecular genotyping of a thalassemia helps to diagnose unexplained microcytosis, and thus prevents unnecessary iron supplementation. PMID- 17132906 TI - De novo isochromosome 18p in a female dysmorphic child. AB - Isochromosome 18p results in tetrasomy 18p. Most of the i(18p) cases reported so far in the literature are sporadic due to de novo formation, while familial and mosaic cases are infrequent. It is a rare chromosomal abnormality, occurring once in every 140,000 livebirths, affecting males and females equally. In the present investigation, we report a de novo i(18p) in a female dysmorphic child. The small metacentric marker chromosome was confirmed as i(18p) in the proband by cytogenetic and FISH analysis [47,XX+i(18p)]. Cytogenetic investigations in the family members revealed normal chromosome numbers, indicating the case as a de novo event of i(18p) formation. It could be due to the somewhat advanced maternal age (32 years) and/or expression of recessive genes in the proband, who is the progeny of consanguineous marriage, which could have led to misdivision and nondisjunction of chromosome 18 in meiosis I, followed by failure in the chromatid separation of 18p in meiosis II and by inverted duplication. PMID- 17132907 TI - Stepwise in-straw dilution and direct transfer using open pulled straws (OPS) in the mouse: a potential model for field manipulation of vitrified embryos. AB - In the present study, mouse blastocysts were employed to investigate the feasibility and efficiency of stepwise in-straw dilution and direct transfer using the open pulled straw (OPS) method. In experiment I, the effects of various vitrification solutions (VS) on embryo survival were examined. After thawing, the expanded blastocyst rates (97.59 and 95.05%) and hatching rates (80.48 and 78.95%) achieved in the EDFS30 [15% ethylene glycol (EG), 15% dimethyl sulfoxide (DMSO), Ficoll, and sucrose] and EFS40 [40% EG, Ficoll, and sucrose] groups were no different from those (96.15% and 83.33%) of the control group. However, the rates in the EFS30 [30% EG, Ficoll, and sucrose] (87.80 and 55.43%) and EDFS40 [20% EG, 20% DMSO, Ficoll, and sucrose] (95.69 and 70.97%) groups were significantly lower than those (96.15 and 83.33%) of the control group (P<0.05). In the experiment II, the effects of the volume of VS in the OPS on the survival of embryos after in-straw thawing were investigated. When the length of the VS in the column was less than 1 cm, the in vitro viability of embryos thawed by stepwise in-straw dilution was no different among the experimental and control groups. The embryos could be successfully thawed by immersing the OPS in 0.5 M sucrose for 3 min and then 0.25 M sucrose for 2 min. In experiment III, the effect of immersion time of the OPS in diluent (PBS) on the viability of vitrified embryos was investigated. After in-straw thawing, OPSs were immersed immediately in 1 ml PBS for 0 to 30 min. When the immersion time of the OPSs in PBS was less than 12 min, in vitro development of the in-straw thawed embryos was no different from that of the controls. In experiment IV, in-straw thawed blastocysts were directly transferred to pseudopregnant mice to examine their in vivo developmental viability. The pregnancy (91.67%) and birth rates (42.42%) of embryos in-straw thawed and directly transferred were no different from those of the unvitrified controls (90.90 and 40%) and embryos thawed by the conventional method (84.61 and 46.94%). These results demonstrate that mouse embryos vitrified with OPS could be successfully thawed by stepwise in-straw dilution and transferred directly to a recipient and that this method might be a model for field manipulation of vitrified embryos in farm animals. PMID- 17132908 TI - Increased Th1/Th2 (IFN-gamma/IL-4) Cytokine mRNA ratio of rat embryos in the pregnant mouse uterus. AB - Somatic cell nuclei can be dedifferentiated in ooplasm from another species, and interspecies cloned embryos can be implanted into the uteri of surrogates. However, no full pregnancies have been achieved through interspecific mammalian cloning. Rat blastocysts transferred into mouse uteri provide a unique model for studying the causes of interspecific pregnancy failure. In this study, intraspecific pregnancy (mouse-mouse) and interspecific pregnancy (rat-mouse) models were established. On Day 9 of pregnancy, the fetoplacental units were separated from the uterine implantation sites and the expression of messenger (m)RNA was quantitated by real-time PCR. We compared the mRNA expression levels of type-1 T helper (Th1) and type-2 T helper (Th2) cytokines, interferon-gamma (IFN-gamma), and interleukin-4 (IL-4) in fetoplacental units between intraspecific and interspecific pregnancy groups. The mRNA expression of IFN gamma in the fetoplacental units of the interspecific pregnancy group was significantly higher than that of the intraspecific pregnancy group (P<0.05). The mRNA expression of IL-4 in the interspecific pregnancy group was significantly lower than that in the intraspecific pregnancy group (P<0.05). We also analyzed the ratio of IFN-gamma/IL-4 mRNA, and an increased IFN-gamma/IL-4 mRNA ratio was observed in the interspecific pregnancy compared with that in the intraspecific pregnancy group. The IFN-gamma and IL-4 mRNA expressions indicate that there is a Th1/Th2 imbalance in the feto-maternal interface of interspecific pregnancies. Bias of Th1 cytokine dominance may be a barrier to reproductive success between species. PMID- 17132909 TI - Expression of porcine FSHbeta subunit promoter-driven herpes simplex virus thymidine kinase gene in transgenic rats. AB - A transgenic rat was established using the construct of porcine FSHbeta subunit promoter, the -852/+10 bp region, fused to a Herpes simplex virus thymidine kinase (HSV-TK) gene. Integration of the transgene was confirmed by PCR of tail DNA. RT-PCR of total RNAs of the pituitary, gonad, cerebellum, liver, kidney, adrenal gland, prostate, and uterus revealed that FSHbeta was only expressed in the pituitary. Analysis of the expression of reporter gene, HSV-TK, using two specific primer sets revealed that different transcripts were present in the pituitary and testis. The transcript initiated at the transcription initiation site of the porcine FSHbeta gene was detected in the pituitary, and another within the TK gene was found in the testis, indicating ectopic testis-specific expression. Immunohistochemistry of the pituitary glands of the transgenic rats for FSH and HSV-TK demonstrated that the FSH-producing cells also produced HSV TK. The results indicated that the -852/+10 bp region of the FSHbeta promoter contains an element(s) that determines the tissue-specific expression. We succeeded in producing FSHbeta promoter-driven HSV-TK transgenic rats and were the first time to do so using an animal other than the mouse. The transgenic rats show male infertility that involves abnormal spermatogenesis. We also observed a decrease in the weight of the testis and epididymis, and both motile and living spermatozoa were absent in the epididymis. Consequently, the FSHbeta-HSV-TK transgenic rat will provide a useful model for studies on FSH function and male infertility. PMID- 17132910 TI - Cellular Localization of Inhibin alpha-subunit, PKB/Akt and FoxO3a proteins in the ovaries of minipigs. AB - Experiments were conducted to examine the cellular localization of inhibin alpha subunit, protein kinase B (PKB/Akt), and FoxO3a proteins in the ovaries of minipigs, Chinese Xiang pigs, by immunohistochemistry. The results indicated that inhibin alpha-subunits were localized in the granulosa cells of follicles at all stages but were not localized in corpora lutea. PKB was localized in the granulosa cells of primordial follicles and in the basal layers of the granulosa cells of preantral and antral follicles, but were not localized in atretic follicles and corpora lutea. FoxO3a was localized in the granulosa cells of follicles at all stages and was extensively localized in the cytoplasma of the luteinized granulosa cells of corpora lutea. Together, the stage- and cell specific expression patterns of inhibin alpha-subunit, FoxO3a, and PKB suggest that these proteins might play potential roles in follicular development, atresia, and luteinization in the minipig. PMID- 17132911 TI - Effects of synchronization of donor cell cycle on embryonic development and DNA synthesis in porcine nuclear transfer embryos. AB - The relationship between donor cell cycle and the developmental ability of somatic cell nuclear transfer (SCNT) embryos has not fully been elucidated. Donor cells that are usually prepared by serum starvation or confluent-cell culture for SCNT represent a heterogeneous population that includes mainly G0 phase cells, other cells in different phases of the cell cycle and apoptotic cells. In this study, we compared the developmental ability of porcine SCNT embryos reconstructed from G0 phase cells (G0-SCNT embryos) and strictly synchronized-G1 phase cells (G1-SCNT embryos), and examined the developmental rates and timing of first DNA synthesis. The G0 phase cells were synchronized by confluent culture, and the G1 phase cells were prepared from actively dividing M phase cells. The G1 SCNT embryos showed a significantly higher (P<0.05) developmental rate to the blastocyst stage per cleaved embryo (59%) than the G0-SCNT embryos (43%). Moreover, initiation of first DNA synthesis and cleavage occurred significantly earlier in the G1-SCNT embryos than in the G0-SCNT embryos. Delay of initiation of first DNA synthesis in the SCNT embryos by aphidicolin resulted in decreased developmental rates to the blastocyst stage without any effect on cleavage rates. Our data demonstrates that synchronized-G1 phase cells can be used as donor cells for SCNT embryos and that earlier initiation of first DNA synthesis may be important for subsequent development of SCNT embryos. The SCNT system using G1 synchronized cells, in terms of their highly uniform and viable cell states, can be useful for studying the reprogramming processes and embryonic development of SCNT embryos. PMID- 17132912 TI - Changes in plasma metabolic hormone concentrations during the ovarian cycles of Japanese black and holstein cattle. AB - The aim of the present study was to investigate the changing profiles of plasma metabolic hormones during the ovarian cycles of beef and dairy cattle. We used 16 non-pregnant, non-lactating Japanese Black beef cattle (6 heifers and 10 cows; parity=2.3 +/- 0.8) and 12 multiparous Holstein dairy cows (parity=3.0 +/- 0.3). Blood samples for hormonal analysis (growth hormone, GH; insulin-like growth factor-I, IGF-1; insulin; and progesterone, P4) were obtained twice weekly for 40 days before artificial insemination for Japanese Black cattle and from 50 to 100 days postpartum for Holstein cows. Luteal phases were considered normal if the P4 concentrations for at least 3 time points over the course of 7 days remained above 1 ng/ml and at least 2 of the time points were above 2 ng/ml. The patterns of the ovarian cycles were classified into two types (normal or abnormal, such as having prolonged luteal phase and cessation of cyclicity) on the basis of the plasma P4 profiles. The plasma concentrations of IGF-1 in both breeds increased transiently during the preovulatory period when the P4 levels were low and decreased to lower levels during the luteal phase when the P4 levels were high. The plasma concentrations of insulin in the 3(rd) week of normal ovarian cycles when the plasma P4 concentration dropped to less than 1 ng/ml were higher than those at other time points in the Japanese Black cattle, but not in the Holstein cows. The plasma concentrations of GH did not change during the ovarian cycle in either breed. In conclusion, the present study indicates that the plasma IGF-1 concentration increases during the follicular phase (low P4 levels) and decreases during the luteal phase (high P4 levels) in non-lactating Japanese Black and lactating Holstein cattle. The results suggest that ovarian steroids, rather than nutrient status, may be related to the cyclic changes in IGF-1 secretion from the liver in cattle. PMID- 17132913 TI - Mutation induction in cultured human cells after low-dose and low-dose-rate gamma ray irradiation: detection by LOH analysis. AB - To study the genetic effects of low-doses and low-dose-rate ionizing radiation (IR), human lymphoblastoid TK6 cells were exposed to 30 mGy of gamma-rays at a dose-rate of 1.2 mGy/hr. The frequency of early mutations (EMs) in the thymidine kinase (TK) gene locus was determined to be 1.7 x 10(-6), or 1.9-fold higher than the level seen in unirradated controls. These mutations were analyzed with a loss of heterozygosity (LOH) detection system, a methodology which has been shown to be sensitive to the effects of radiation. Among the 15 EMs observed after IR exposure, 8 were small interstitial-deletion events restricted to the TK gene locus. However, this specific type of event was not found in unirradiated controls. Although these results were observed under the limited conditions, they strongly suggest that the LOH detection system can be used for estimating the genetic effects of a low-dose IR exposure delivered at a low-dose-rate. PMID- 17132914 TI - Repair of potentially lethal damage in normal cells and ataxia telangiectasia cells; consideration of non-homologous end-joining. AB - When cell lines are held in a quiescent state after irradiation, survival rates are greater than those from cells that are stimulated to grow immediately after irradiation. These differences in survival rates correspond to rates of potentially lethal damage repair. The effects of confluent holding recovery after gamma-irradiation were investigated using normal human fibroblasts (AG1522) and ataxia telangiectasia fibroblasts (GM02052). Calyculin-A-induced premature chromosome condensation and fluorescent in situ hybridization were applied to study G2/M chromosomal aberrations. Survival results indicated normal capacity for PLDR in AG1522 cells but that PLDR was extremely compromised in GM02052 cells. The chromosomal aberration frequency decreased when AG1522 cells were allowed to repair for 24-h, whereas 24-hour incubation had little effect on the aberration frequency in GM02052 cells. Since the main mechanism for dsbs repair during G0/G1 phases of the cells cycle involve the non-homologous end-joining (NHEJ) process, our study indicates that for AG1522 cells the NHEJ repair process is more likely to induce accurate chromosome repair under quiescent G0 conditions than proliferating G1 phase, while in GM02052 cells the fidelity of NHEJ is similarly defective at either cell cycle phase. Reduced fidelity of NHEJ may be responsible for PLDR defect and its hyper-radiosensitivity in A-T cells. PMID- 17132915 TI - The chemopreventive flavonoid apigenin confers radiosensitizing effect in human tumor cells grown as monolayers and spheroids. AB - Apigenin, a common dietary flavonoid present in many fruits and vegetables, is a nonmutagenic chemopreventive agent. In the present study, we investigated the effect of apigenin on the radiosensitivity of SQ-5 cells, which are derived from a human lung carcinoma. Actively growing cells were incubated for 16 h at 37 degrees C in medium containing 40 muM apigenin. The cells were then irradiated with X-rays and incubated with apigenin for a further 8 h. Radiosensitivity was assessed using a clonogenic assay. Apoptosis and necrosis were assessed using acridine orange/ethidium bromide double staining. Cells incubated with apigenin exhibited significantly greater radiosensitivity and apoptosis levels than cells not incubated with apigenin. Protein levels were measured by Western blotting. Incubation with apigenin increased protein expression of WAF1/p21 and decreased protein expression of Bcl-2. Furthermore, apigenin sensitized SQ-5 spheroids (cell aggregates growing in a three-dimensional structure that simulate the growth and microenvironmental conditions of in vivo tumors) to radiation. Thus, apigenin appears to be a promising radiosensitizing agent for use against human carcinomas. PMID- 17132916 TI - [A case of colonic stricture induced by ulcerative colitis]. PMID- 17132917 TI - [Hepatic fibrogenesis]. AB - In acute injury, liver recovers completely without any scarring change or complication. However, large portion of liver is changed into fibrotic state by excessive production of extracellular matrix (ECM) under chronic injury. Excessive production of ECM results in hepatic fibrosis and repeated process of hepatic fibrosis progress into liver cirrhosis. Liver cirrhosis is an irreversible and terminal state of chronic liver disease and one of the major causes of death in Korea. To block the progression to liver cirrhosis, various studies in the field of virology and immunology have been proceeded. Recently, studies on the hepatic fibrogenesis have progressed with the development of molecular biology. Hepatic stellate cells (HSC) play a key role in the pathogenesis of hepatic fibrosis by producing ECM. The degree of hepatic fibrosis depends on the proliferation and activation of HSC and increased net production of collagen. Therefore, inhibition of HSC activation is one of the main ways to block the progression of hepatic fibrosis. Many kinds of factors such as oxidative stress, acetaldehyde, ascorbic acid, transforming growth factor-beta (TGF-beta) and carbon tetrachloride (CCl4) have been reported to activate HSC and stimulate collagen gene expression. Although there are no definite and effective antifibrogenic agents, possible candidates are antioxidants, interferon, retinoids such as beta-carotene, flavonoids, renin-angiotensin system inhibitors and peroxisome proliferator activated receptor-gamma (PPAR-gamma) agonists. We tried to evaluate the charateristics of HSC in order to develop agents that inhibit hepatic fibrogenesis. PMID- 17132918 TI - [Risk of postoperative infection in patients with inflammatory bowel disease]. AB - BACKGROUND/AIMS: The clinical course of patients with inflammatory bowel disease (IBD) frequently leads to the use of immunosuppressants and immunomodulators. We investigated the risk of postoperative infection in patients with IBD undergoing elective bowel surgery and whether the use of corticosteroid (CS) and/or 6 mercaptopurine/ azathioprine (6-MP/AZA) before surgery was associated with the increased risk of postoperative infection. METHODS: Patients who were diagnosed as Crohn's disease (n=25) or ulcerative colitis (n=19) and underwent elective bowel surgery between 1986 and 2005 were identified. Medical records were retrospectively analyzed including age, sex, duration of disease, indication for surgery, duration of surgery, type of surgery, type of postoperative infection, admission period, usage of CS and 6-MP/AZA, and preoperative laboratory values. There were 27 patients receiving CS alone, 6 patients receiving 6-MP/AZA alone or with CS, and 16 patients receiving neither CS nor 6-MP/AZA. RESULTS: There were 17 postoperative infections (38.6%) among IBD patients who had undergone surgery and wound infection was the most common type of infection (76.5%). In IBD patients, patients receiving CS had higher postoperative infection rate than those patients receiving neither CS nor 6-MP/AZA (p=0.039). Patients receiving CS in conjunction with 6-MP/AZA did not have significantly higher postoperative infection rate than those with CS only (p=0.415). CONCLUSIONS: Preoperative use of CS in patients with IBD is associated with the increased risk of postoperative infections. Addition of 6-MP/AZA in patients receiving CS does not increase the risk of postoperative infections. PMID- 17132919 TI - [Comparative usefulness of erythrocyte sedimentation rate and C-reactive protein in assessing the severity of ulcerative colitis]. AB - BACKGROUND/AIMS: Although erythrocyte sedimentation rate (ESR) is included as a laboratory parameter in Truelove and Witts' classification, C-reactive protein (CRP) is also used for severity assessment in ulcerative colitis (UC). Frequently, the discordance between ESR and CRP is observed in clinical practice. The aim of this study was to determine which parameter is more related with clinical activity in UC patients. METHODS: A total of 155 patients with UC were identified from January 2004 to March 2005. Their medical records were reviewed within these patients, a total of 541 assessments of disease activity were made. Correlation of clinical activity and laboratory tests were evaluated by Pearson's correlation coefficient. RESULTS: Pearson's correlation coefficients of ESR and CRP with clinical symptoms were 0.376 and 0.258, respectively. The correlation coefficient between ESR and CRP was 0.403 (p=0.000). A total of 131 (24.2%) assessments revealed discordance between ESR and CRP. When discordance occurred, the correlation coefficients with clinical symptoms were 0.338 for ESR (p=0.000) and 0.034 for CRP (p>0.01). Dividing discordant patients into high ESR/low CRP group and low ESR/high CRP group, the coefficients were 0.420 for ESR and 0.226 for CRP in high ESR/low CRP group, and 0.333 for ESR and 0.068 for CRP in low ESR/high CRP group. CONCLUSIONS: The correlation analysis indicates that ESR appears to be a more reliable laboratory parameter of disease activity than CRP in assessing the severity of UC. In particular, when the level of ESR and CRP is discordant, ESR is more useful in assessing the disease activity in UC patients. PMID- 17132920 TI - [Usefulness of serum alpha-fetoprotein (AFP) as a marker for hepatocellular carcinoma (HCC) in hepatitis C virus related cirrhosis: analysis of the factors influencing AFP elevation without HCC development]. AB - BACKGROUND/AIMS: Serum alpha-fetoprotein (AFP) is frequently used for the diagnosis of hepatocellular carcinoma (HCC). Most available data concerning AFP came from studies of patients with chronic hepatitis B or mixed etiologies. Studies concerning the diagnostic value of AFP for HCV-related liver cirrhosis (LC) are limited. We evaluated the factors influencing AFP elevation in the absence of HCC and analyzed the diagnostic value of serum AFP in HCC surveillance of HCV-related LC patients. METHODS: We enrolled 55 patients of HCV-related LC with HCC and 62 patients without HCC as a case-control study were analyzed. The sensitivity and specificity were calculated and the clinical and biochemical factors influencing serum AFP levels. RESULTS: The sensitivities and specificities of serum AFP for the detection of HCC in HCV-related LC were 72.7% and 59.7% for AFP>or=20 ng/mL, and 47.3% and 92.5% for AFP>or=100 ng/mL, respectively. Elevated serum AST was independently associated with elevated serum AFP level in HCV-related LC. In cases of ASTor=100 ng/mL for the diagnosis of HCC was 100%. However, in case of AST>2 ULN, the specificity was 85.0% for AFP>or=100 ng/mL and 95.0% for AFP>or=200 ng/mL. CONCLUSIONS: Serum AST levels influence serum AFP level in HCV-related LC. In cases of AST2 ULN. PMID- 17132921 TI - [Methylation pattern of DNA repair genes and microsatellite instability in hepatocelluar carcinoma]. AB - BACKGROUND/AIMS: Epigenetic silencing of DNA repair genes, O6-methylguanine-DNA methyltransferase (MGMT), hMLH1 and hMSH3, by promoter hypermethylation have been observed in various cancers. However, the relationship between hypermethylation of DNA mismatch repair genes and microsatellite instability (MSI) has not been studied in hepatocellular carcinoma (HCC) associated with cirrhosis. METHODS: We investigated the methylation pattern of CpG islands of 3 genes using methylation specific PCR (MSP) and MSI in 40 patients with paired hepatocellular carcinoma and associated cirrhosis. RESULTS: hMSH3 and MGMT were the most methylated genes in both cirrhosis (70% and 68%, respectively) and HCC (75% and 73%, respectively). The methylation of hMLH1 was rarely found in both cirrhosis (8%) and HCC (5%). Gene promoters methylated in cirrhosis were also methylated in HCC with the exception of 9 cases found to be methylated either in cirrhosis or HCC. Of 40 cases of HCC associated with cirrhosis, three had MSI-positive phenotype in which two were MSI-low and one was MSI-high. One MSI-positive phenotype was present both in cirrhosis and in HCC, while two were only in HCC. There was no significant correlation between aberrant DNA methylation of mismatch repair genes and MSI status in HCC associated with cirrhosis. Immunohistochemical expressions of hMLH1, MGMT, and hMSH3 proteins were present in 16 (40%), 6 (15%), and 11 (28%) of 40 cases of HCC respectively. There was no significant correlaton between the aberrant DNA methylation of mismatch repair genes and clinical characteristics such as histological differentiation, postoperative recurrence and mortality. CONCLUSIONS: The methylation of MGMT and hMSH3 among DNA repair genes are frequent, but those of hMLH1 and MSI is very rare in both cirrhosis and HCC. There is no significant correlation between the methylation of DNA repair genes and clinical characteristics of HCC. PMID- 17132922 TI - [Outcome of intensive medical treatments in patients with infected severe necrotizing pancreatitis]. AB - BACKGROUND/AIMS: Infection of pancreatic necrosis is one of the leading cause of death in patients with severe necrotizing pancreatits. Because of high mortality rate up to 50%, immediate surgical debridement including pancreatectomy is recommended. However, early surgical treatment still showed high mortality rate and better treatment strategy is required. This study was conducted to evaluate the outcomes of early intensive non-surgical treatments in patients with infected necrotizing pancreatitis. METHODS: This study was based on retrospective analysis of 71 patients with acute severe necrotizing pancreatitis (APACHE II score>or=8, or Ranson's score>or=3, and pancreatic necrosis on CT scan), who were admitted to medical center during past 16 years. Infection of pancreatic necrosis was confirmed by fine needle aspiration, and early intensive medical treatments comprised of prophylactic antibiotics coverage, fluid resuscitation, organ preserving supportive measures, and percutaneous catheter drainage were carried out. RESULTS: Among the enrolled patients, infections were suspected in 46 patients, but fine needle aspirations were done only in 32 patients. In 21 patients, infections of necrotic tissue were confirmed by bacteriology, while other 11 patients showed no evidence of bacterial growth. Of 21 patients with infected necrosis, initial surgical interventions were performed in 2 patients, while initial medical treatments were performed in 19 patients. The success rate of medical treatment group in infected necrotizing pancreatitis was 79% (15/19). The mortality rate of medical treatment group and surgical treatment group was 5% (1/19) and 50% (1/2). CONCLUSIONS: Early intensive medical treatment seems to be a good therapeutic strategy, even if the infection has developed in pancreatic necrosis. Further prospective randomized studies are required to confirm this finding. PMID- 17132923 TI - [Prevalence and risk factors of gallbladder polyp in adults living in Daegu and Gyeongbuk provinces]. AB - BACKGROUND/AIMS: Although there are various reports on the prevalence of gallbladder (GB) polyp according to the race or area, there is no report on Daegu and Gyeongbuk provinces. Therefore, we investigated the prevalence and related risk factors of GB polyp in health screening population of Daegu and Gyeongbuk provinces. METHODS: Of 28,911 adult health screening examinees from January 1997 to May 2003, age and gender-matched 8,008 (3,896 men, 4,112 women) were randomly selected to evaluate the prevalence. Independent risk factors were analyzed by comparing various variables between the GB polyp positive and negative groups. RESULTS: The prevalence of GB polyp was 2.2% (175/8,008). Those were 2.7% (104/3,896) among men and 1.7% (71/4,112) among women. Independent risk factors related with GB polyp were male gender (OR 1.434, p<0.001), obesity (OR 1.237, p=0.024), and hepatitis B carrier (OR 1.888, p<0.001) CONCLUSIONS: Prevalence of GB polyp in Daegu and Gyeongbuk provinces is 2.2%. Male, obesity, and hepatitis B carrier may be the risk factors related with GB polyp in this region. PMID- 17132924 TI - [A case of pancreatic somatostatinoma]. AB - Somatostatinoma is a rare neoplasm usually arising from the pancreas and duodenum which typically presents with indolent, nonspecific symptoms in the absence of systemic neuroendocrine manifestations that characterize somatostatinoma syndrome. It accounts for less than 1% of all gastrointestinal endocrine tumors with an annual incidence of 1 per 40 million. It is often associated with regional and/or portal metastasis at the time of diagnosis, and complete tumor resection is possible only in 60% to 70% of cases. We experienced a case of pancreatic somatostatinoma recently. A 51-year-old woman presented with right upper quadrant abdominal pain and loose stool for one month. A hypermetabolic lesion in the pancreatic head was detected on positron emission tomography-CT (PET-CT) scan. The tumor was resected by pylorus preserving pancreaticoduodenectomy. Immunohistochemical staining of the tumor tissue exhibited diffuse positivity for somatostatin, but was negative for insulin and glucagon. Herein, we report a case of pancreatic somatostatinoma diagnosed postoperatively. PMID- 17132925 TI - [Undifferentiated carcinoma with osteoclast-like giant cells of the pancreas]. AB - Undifferentiated carcinoma with osteoclast-like giant cells is a rare neoplasm of exocrine pancreas. Till recently, some cases have been reported, however histogenesis of the tumors are controversial and their characteristic findings have not been described yet. Thirty five-year-old men and 75-year-old men were presented with upper abdominal pain and a palpable mass. On computed tomography, one case showed a well enhancing solid tumor with low density and the other was showed a mainly cystic tumor with peripheral enhancement in the body and tail of the pancreas. One case accompanied multiple metastatic liver masses with subhepatic lymph node enlargement. Tumor staining was seen on angiography. Biopsy and pancreatectomy were performed. Pathological findings revealed tumors composed of neoplastic spindle shaped or pleomorphic large cells with scattered non neoplastic osteoclast-like giant cells. In one case, there were small foci of adenocarcinoma components in the periphery of the tumor. On immunohistochemical stain, neoplastic cells showed focal positivity for epithelial membrane antigen and vimentin. Tumors were diagnosed as undifferentiated carcinoma with osteoclast like giant cells. We report these rare cases with a review of literature. PMID- 17132926 TI - [A case of primary adenosquamous carcinoma of the liver with formation of colonic fistula]. AB - Primary adenosquamous carcinoma of the liver is generally considered as an extremely rare subtype of cholangiocarcinoma. It has been reported mostly in a form of case studies. As far as we know, there was only one case report on tumor related with biliary fistula. Recently, we experienced a case of primary adenosquamous carcinoma of liver with a formation of tumor-colonic fistula. A 54 year-old man was transferred to our hospital due to liver mass detected by abdominal ultrasonogram. Dynamic computed tomogram of liver showed a large irregular hypodense mass without rim enhancement in right lobe of liver and also suggested a fistula formation between the tumor and hepatic flexure of right colon. Colonoscopic examination showed a large colonic wall defect in hepatic flexure and a friable, nodular mucosa around the defected colonic wall. Extended right lobectomy and right hemicolectomy were done. Microscopically, the tumor was composed of squamous cell carcinoma mainly with foci of the adenocarcinoma component. PMID- 17132927 TI - [A case of primary jejunal adenocarcinoma diagnosed by enteroscopy using pediatric colonoscope]. AB - A 39-year-old man presented with dizziness and melena for 2 months. Abdominal CT scan showed constrictive wall thickening with enhancement and proximal loop dilatation of the jejunum. On endoscopic examination, there was large amount of bile stained fluid in duodenum. Enteroscopy using pediatric colonoscope demonstrated an encircling mass with obstruction approximately 20 cm distal to the ligament of Treitz. Endoscopic jejunal biopsy showed moderately differentiated adenocarcinoma. Small intestinal adenocarcinoma is uncommonly encountered in clinical practice. Because small intestine is relatively inaccessible via routine endoscopy, diagnosis of small intestinal neoplasm is often delayed for several months after the onset of symptoms. Most of the patients are diagnosed in advanced stage. Therefore, when a small bowel neoplasm is suspected, enteroscopy is the most useful study. If enteroscope is not available, enteroscopy using pediatric colonoscope may permit earlier preoperative diagnosis. We report a case of primary jejunal adenocarcinoma diagnosed by endoscopic biopsy using pediatric colonoscope. PMID- 17132928 TI - [Intensive medical treatments of infected severe necrotizing pancreatitis]. PMID- 17132930 TI - RNA repair--the latest addition to the toolbox for macromolecular maintenance. AB - All organisms must keep their DNA intact and free of lesions to obtain faithful transmission of genetic information, and many cellular proteins are involved in genome maintenance. However, also other macromolecules may be subject to repair. Deleterious lesions in proteins can be reversed by several mechanisms, and this appears to be particularly important in cell types where protein turnover is low. In addition, recent studies indicate that RNA is also repaired. AlkB proteins, which were initially found to repair chemically alkylated DNA, can also remove alkyl lesions from RNA, and, moreover, provide functional reactivation of damaged tRNA and mRNA. Here, recent findings on RNA repair are discussed. PMID- 17132931 TI - A unified mode of epigenetic gene silencing: RNA meets polycomb group proteins. AB - Recently, an essential role for RNA in the epigenetic silencing of genes packaged within heterochromatin in animals has been recognized. The RNA appears to be involved in targeting chromatin remodeling activity to a specific locus and in later maintaining the repressed state of the gene. Epigenetic silencing of Hox cluster genes by the Polycomb group proteins also involves the formation of a stably inherited repressive chromatin structure. Recent studies of the C. elegans PcG gene sop-2 reveled an evolutionarily conserved property of PcG proteins in the binding of RNA, suggesting an important role for RNA in PcG-mediated Hox gene repression. PMID- 17132932 TI - HuR: post-transcriptional paths to malignancy. AB - The RNA-binding protein HuR regulates the stability and translation of target mRNAs. While no HuR mutations have been found in cancer, a link between HuR and malignant transformation has been suggested in cancers of the breast, colon, lung and ovary. We describe a paradigm consistent with a central role of HuR in oncogenesis. PMID- 17132933 TI - Selection by phage display of peptides targeting the HIV-1 TAR element. AB - As transcription regulatory element, the HIV-1 TAR RNA element is a promising target to inhibit viral replication; indeed, ligands of TAR RNA could prevent the transcription trans-activation process. Phage display in vitro selection was undertaken to select peptidic ligands of TAR RNA. In preliminary experiments, the selection was performed in a magnesium rich buffer (3 mM), but only phages targeted to plastic wells or streptavidin emerged; in addition, a "super infectious" phage present in the New England Biolabs library (SVSVGMKPSPRP) selected by others with different targets was cloned, due to a high amplification potential. In contrast, the absence of magnesium or an increasing magnesium concentration (0 to 0.5 mM) led to phage selection with 57 amino acid peptides. K(D)s of 420-550 nM were measured by filter binding assays; a significant specificity was obtained when TAR target was compared with unrelated RNA targets. Surprisingly, the binding of selected peptides does not depend on the magnesium concentration. PMID- 17132934 TI - RNA damage in human atherosclerosis: pathophysiological significance and implications for gene expression studies. AB - RNA damage is a poorly examined field in biomedical research. Potential triggers of RNA damage as well as its pathophysiological implications remain largely unknown. Here we summarize recent evidence that loss of RNA integrity and 7,8 dihydro-8-oxo-2'-guanosine (8-oxoG) oxidative RNA modifications frequently occur in advanced human atherosclerotic plaques. At least two features of advanced human plaques, namely oxidative stress and intraplaque hemorrhage followed by iron deposition, may be involved in the process of RNA degradation. Although speculative, RNA damage may lead to destabilization and rupture of atherosclerotic plaques by interfering with protein synthesis and stimulation of cell death. Moreover, RNA damage may affect in vitro transcript quantification, thereby influencing data from gene expression studies. PMID- 17132935 TI - Inhibition of HIV-1 replication by siRNA targeting conserved regions of gag/pol. AB - Short interfering RNAs (siRNAs) targeting HIV-1gag, vif, tat, rev and host CD4 and CCR5 have been reported to inhibit HIV replication. However, the sequence divergence of HIV and the concentration dependence of siRNA activity represent significant challenges to RNAi mediated inhibition. To determine the parameters of RNAi in suppression of HIV-1 we screened seven siRNA candidates targeting highly conserved regions of gag/pol, based on target site GC content, for antiviral activity at varying concentrations. Only two of these inhibited CA-p24 production more than 50%, 2064 and 2161. Activity varied with concentration, with 100 nM producing optimal suppression. Requirements for target sequence conservation and activity over a range of concentrations may severely limit the number of siRNA candidates for therapeutic development. PMID- 17132936 TI - Selecting effective siRNA target sequences for mammalian genes. AB - Short interfering RNA (siRNA) has been widely used for studying gene functions in mammalian cells but varies markedly in its gene-silencing efficacy in mammalian genes. The recently reported guidelines for selecting effective siRNA target sequences are not always useful for selecting highly effective siRNA sequences for many other mammalian genes because there are only a few consistencies among them. Hypothesizing that the positional nucleotide occurrence trends play an important role in effective gene-silencing, we examined 361 effective siRNA sequences from 227 different mammalian cDNAs in the literature and found got several nucleotide features different from the ones used in the previous guidelines. Here we first explain the problems concerning the previous guidelines from the qualitative and quantitative points of view. Then after clarifying the requirements for effective siRNA designs, we describe a new method based on a gene degradation measure defined by positional features of specific significant nucleotides. Testing the method on human cyclin B1 confirmed that it selected highly effective gene-silencing sequences and also indicated that it would be useful for other genes. It will therefore be useful for selecting new siRNA target sequences for mammalian genes. PMID- 17132937 TI - Lost once, the Fragile X Mental Retardation protein is now back onto brain polyribosomes. AB - The Fragile X Mental Retardation protein (FMRP) is an RNA-binding protein and its absence leads to the Fragile X syndrome, the most common form of inherited mental retardation. Because it has been acknowledged for a long time that FMRP is associated with polyribosomal mRNPs in all non-neuronal cellular systems studied so far, it is thought that it regulates translation in neurons also; however, its exact function remains elusive. Recently, it has been reported that, contrary to non-neuronal cells, brain FMRP is not associated with the translation machinery, but is part of repressed small RNP complexes excluded from polyribosomes.(27) To elucidate this puzzling result, Stefani et al.(17) and Khandjian et al.(32) have optimized methods to analyze brain polyribosomes and now provide definitive evidence for the association of FMRP with brain polyribosomes. In addition, the data presented in these two reports clearly indicate that FMRP's function resides at the translation control level. PMID- 17132938 TI - Natural antisense transcripts. AB - The sequencing of whole genomes and the subsequent annotation of cDNAs revealed that about 20% of human and mouse genes overlap resulting in potential pairs of sense and antisense transcripts. An increasing number of experimentally identified antisense transcripts concur with this predication. Characterization of overlapping transcripts in various species indicates that this form of RNA mediated gene regulation represents a widespread phenomenon. However, the physiological relevance of natural antisense transcripts remains obscure. Genomic studies suggest that duplex formation between sense and antisense is required for biological function. Antisense transcripts play an established role in imprinting and X-chromosome inactivation and genomic rearrangements as observed in B and T leukocytes. Only a relatively small percentage of the predicted antisense transcripts are related to these biological phenomena that are also related to mono-allelic expression. Consequently, there are at least two categories of natural antisense transcripts that show significant differences with regard to their biological function as well as the potential mechanisms involved. PMID- 17132939 TI - A link between transcription and mRNP quality in Saccharomyces cerevisiae. AB - The transcription machinery plays a direct role in the assembly of messenger ribonucleoprotein particles (mRNPs), contributing to the loading of proteins onto nascent transcripts. Such mRNP biogenesis is linked to the THO complex that operates at the boundary between transcription and nuclear export. Early mRNP assembly events are subject to surveillance by the nuclear exosome that retains, and degrades, aberrant mRNAs. A yeast strain that carries deletions of Hpr1p and Rrp6p of the THO complex and the nuclear exosome, respectively, grows slowly, possibly due to lack of Rrp6p-dependent mRNA quality control. We selected a number of spontaneous revertant strains from the slow growth phenotype. Interestingly, transcriptional activity was reduced in all revertants. These data corroborate earlier findings that transcriptional down regulation improves growth of cells containing a crippled mRNP formation/surveillance system and illustrates the impact transcriptional activity can have on early mRNP assembly. PMID- 17132940 TI - QUAKING KH domain proteins as regulators of glial cell fate and myelination. AB - The quaking viable (qk(v)) mice have attracted attention because of their characteristic tremor caused by their dysmyelination. In the central nervous system, qk(v) mice fail to develop mature myelinating oligodendrocytes and display uncompacted myelin. The genetic defect in the qk(v) mice prevents the proper expression of alternatively spliced KH-type QKI RNA binding proteins. Thus qk(v) mice provide a unique animal model linking RNA binding proteins to defects in oligodendrocyte cell fate and myelination. The fact that QKI proteins are modified post-translationally makes them Signal Transduction Activiators of RNA (STAR) proteins. We have used a gain-of-function approach with the ectopic expression of the separate QKI isoforms using adenoviruses and retroviruses to determine their separate roles in cell fate and myelination. Herein, we discuss the recent advances in characterizing the QKI KH-type proteins as glial cell fate and myelin egulators. PMID- 17132941 TI - Human mitochondrial diseases associated with tRNA wobble modification deficiency. AB - A growing number of mutations in mitochondrial (mt) tRNA genes have been found to associate with human mitochondrial diseases. Our previous analysis of mutant mt tRNAs isolated from cells derived from patients with mitochondrial diseases revealed the lack of a post-transcriptional taurine-modification at the anticodon wobble uridine in two mt tRNAs bearing typical pathogenic mutations: mt tRNA(Leu(UUR)) with either the MELAS 3243 or 3271 mutation and mt tRNA(Lys) with the MERRF 8344 mutation. We here summarize our recent studies that clarify the molecular basis of the defective mitochondrial translation caused by this wobble modification deficiency. The MERRF mt tRNA(Lys) lacking the wobble modification cannot translate either of its codons (AAA and AAG), while the translational activity of MELAS mt tRNA(Leu(UUR)) lacking wobble modification is more depressed in decoding of UUG codon than UUA codon. These findings suggest that the wobble modification deficiency plays a primary role in the molecular pathogenesis of the MELAS and MERRF mitochondrial diseases. PMID- 17132942 TI - Molecular genetics of spinocerebellar ataxia type 8 (SCA8). AB - Spinocerebellar ataxias (SCAs) belong to a group of autosomal dominant, late onset neurodegenerative disorders characterized by slowly progressive ataxia that eventually leads to severe gait, speech, coordination and sensory loss. The majority of these diseases result from expanded polyglutamine tracts in the encoded protein as seen in SCA1, SCA2, SCA3, SCA6, SCA7 and Dentatorubral Pallidoluysian Atrophy (DRPLA). However, two novel forms of SCAs, SCA8 and SCA12, are associated with trinucleotide repeat expansions in non-translated regions of the genes. In the case of SCA8, the CUG expansion occurs at the 3' end of a processed non-coding RNA. While understanding of how expanded polyglutamine tracts compromise or alter protein function has advanced rapidly in the last five years, understanding of how trinucleotide repeat expansions alter the function of the non-coding SCA8 RNA and lead to human disease remains quite limited. Encouragingly, as discussed in this review, recent studies from murine and Drosophila models have provided new insights into both the cellular context in which SCA8 normally operates and the potential role of CTG expansion in the disease. Continued exploration of these genetically tractable model systems will further illuminate the biology underlying SCA8 disease, ultimately providing the necessary foundation on which to develop effective therapeutic interventions. PMID- 17132943 TI - tmRNA decreases the bactericidal activity of aminoglycosides and the susceptibility to inhibitors of cell wall synthesis. AB - Trans-translation is a process that recycles ribosomes stalled on problematic mRNAs. tmRNA, coded by the DeltassrA gene, is a major component of trans translation. Bacteria lacking tmRNA are more sensitive to several inhibitors of protein synthesis when compared to a wild type strain. We measured bacterial growth of the DeltassrA and wild type strains in Escherichia coli in the presence of 14 antibiotics including some that do not target protein synthesis. Both the optical density of the bacterial cultures and the number of viable cells were monitored. For the ribosome-targeted antibiotics, sensitization was observed on erythromycin, chloramphenicol, kanamycin, puromycin and streptomycin. Minor or no effects were observed with clindamycin, tetracycline and spectinomycin. Surprisingly, the DeltassrA strain is more sensitive than wild type to inhibitors of cell wall synthesis: fosfomycin and ampicillin. No growth difference was observed on drugs with other target sites: ofloxacin, norfloxacin, rifampicin and trimethoprim. Sensitization to antibiotics having target sites other than the ribosome suggests that trans-translation could influence antibiotic-induced stress responses. In trans-translation-deficient bacteria, cell death is significantly enhanced by the two aminoglycosides that induce translational misreading, streptomycin and kanamycin. PMID- 17132944 TI - Inhibition of HIV-1 replication by an improved hairpin ribozyme that includes an RNA decoy. AB - An anti-Tat hairpin ribozyme and a TAR RNA decoy were combined in one molecule. The chimeric molecule strongly inhibited HIV-1 replication (measured as changes in p24 levels in viral replication assays). The inhibitory action of the ribodecozyme (85%) was significantly greater than that shown by ribozyme and a non-catalytic variant carrying the functional decoy RNA domain (55% and 35%, respectively). This represents a significant improvement of the inhibitory efficiency of the ribozyme, suggesting there is an additive inhibitory effect on HIV-1 replication by the catalytic and decoy domains. This strategy could be used to create new inhibitor RNAs with enhanced in vivo performance. PMID- 17132945 TI - Arrangement of mRNA 3' of the A site codon on the human 80S ribosome. AB - Positioning of mRNA 3' of the A site codon was studied with the use of short mRNA analogues carrying a UUU triplet at the 5'-termini and a perfluorophenylazide group at either the N7 atom of the guanosine or the C5 atom of the uridine 3' of the triplet. Modified nucleotides were directed to positions +7, +9 or +12 with respect to the first nucleotide of the P site codon by tRNA(Phe) cognate to the UUU triplet targeted to the P site. Mild UV-irradiation resulted in cross-linking of the mRNA analogues to the 18S rRNA and to 40S proteins, the yield of cross linking depending on the nature of the mRNA nucleotide bearing the modified group and its position on the ribosome. In addition, the yield of cross-linking to the 18S rRNA decreased strongly when the modified nucleotide was moved from position +7 to position +12. All the mRNA analogues cross-linked to the 18S rRNA nucleotides that had been found earlier at the decoding site, namely, to the invariant dinucleotide A1824/A1825 and the variable A1823 in the 3'-minidomain of the 18S rRNA, and also to the invariant nucleotide C1698 in the 3'-minidomain and the conserved region 605-620 in the apical region of helix 18 in the 5'-domain. The results indicate that (1) the mRNA makes a sharp turn between positions +6 and +7, (2) the triplet immediately 3' of the A site codon neighbors the 18S rRNA and proteins, and (3) the codon 3' of the triplet mentioned is surrounded mainly by proteins. PMID- 17132946 TI - Silver: an old wine in a new bottle. PMID- 17132950 TI - Buruli ulcers: an emerging health problem in Ghana. PMID- 17132951 TI - Support service initiative: terms, definitions, and patient care. PMID- 17132952 TI - Revised coding guidelines for surgical dressings. PMID- 17132953 TI - Never give up on your dreams. PMID- 17132955 TI - Use of high-resolution, high-frequency diagnostic ultrasound to investigate the pathogenesis of pressure ulcer development. AB - OBJECTIVES: To investigate the pathogenesis of pressure ulcers utilizing high resolution ultrasound and to explore the utility of this technology for the detection of incipient pressure ulcers prior to visual clinical signs. DESIGN: An observational prospective study comparing high-resolution ultrasound images obtained from 119 long-term-care facility residents determined to be at risk for pressure ulcer development (Braden Scale score of 18 or less) with images obtained from 15 healthy volunteers (medical students and medical residents). Common pressure ulcer sites were scanned, including the heels, sacrum, and ischial tuberosity. SETTING: A medical center and a long-term-care facility. INTERVENTION: Anatomic sites universally accepted as at risk for pressure ulcer development were scanned using high-resolution ultrasound; the sites did not have visual evidence of skin breakdown. The images obtained from the long-term-care facility residents were compared with images considered normal that were obtained from healthy volunteers. In addition, documentation of the clinical assessment finding for erythema was reviewed, recorded, and compared with the high resolution ultrasound finding for each specific site. MEASUREMENTS: The images obtained were classified as not readable, normal, or abnormal. The images classified as abnormal were further classified by depth of abnormal finding: pattern 1 (deep) or pattern 2 (superficial). The images classified with the abnormal finding pattern 1 (deep) were further classified and subdivided by anatomic location of abnormal finding(s): subgroup 1, abnormal findings in the subdermal area only; subgroup 2, subdermal and dermal abnormal findings; and subgroup 3, subdermal, dermal, and subepidermal edema. Pattern 2 (superficial) included images with abnormal findings limited to the dermal/epidermal junction. RESULTS: 630 (55.3%) of the images obtained from the long-term-care residents were different from the images obtained from the healthy volunteers. The healthy volunteers' images classified as normal had the expected ultrasound findings for homogeneous pattern of ultrasound reflections, allowing for visualization of various skin layers (epidermis, superficial papillary dermis, deep reticular dermis, and hypodermis) and subcutaneous tissue (subdermal). However, many images (55.3%) obtained from the residents at risk for pressure ulcer development had patterns where areas within the various skin layers were not visible, interrupted by areas indicative of fluid or edema. Moreover, most images (79.7%) with abnormal ultrasound patterns did not have documentation of erythema. CONCLUSION: High-resolution ultrasound is an effective tool for the investigation of skin and soft tissue changes consistent with the documented pathogenesis of pressure ulcers. A progressive process for pressure ulcer development from deep subdermal layers to superficial dermal then epidermal layers can be inferred. Dermal edema was only present with subdermal edema. In other words, there was never evidence of dermal edema in the absence of subdermal edema. A better understanding of the pathogenesis of pressure ulcers through the use of high-resolution ultrasound to detect soft tissue damage and edema before visible clinical signs could lead to earlier and more focused pressure ulcer prevention programs, resulting in reduced pain and suffering for improved patient quality of life and wound care cost savings. PMID- 17132956 TI - Debridement: a vital component of wound bed preparation. AB - PURPOSE: To provide practitioners with an overview of debridement and its role in wound healing. TARGET AUDIENCE: This continuing education activity is intended for physicians and nurses who assess and treat wounds. OBJECTIVES: After reading the article and taking the test, the reader should be able to: 1. Discuss the physiologic process of wound healing and the rationale for debridement. 2. Describe evidence-based indications for and methods of debridement. PMID- 17132959 TI - The end of AD. Part 3. PMID- 17132961 TI - In search of the molecular basis of memory loss in Alzheimer disease. PMID- 17132962 TI - Normal and abnormal tau neurobiology. PMID- 17132963 TI - The natural history of Alzheimer disease dissected through multiphoton imaging of transgenic mice. PMID- 17132964 TI - The Uniform Data Set (UDS): clinical and cognitive variables and descriptive data from Alzheimer Disease Centers. AB - A Clinical Task Force, composed of clinical leaders from Alzheimer's Disease Centers (ADC), was convened by the National Institute on Aging to develop a uniform set of assessment procedures to characterize individuals with mild Alzheimer disease and mild cognitive impairment in comparison with nondemented aging. The resulting Uniform Data Set (UDS) defines a common set of clinical observations to be collected longitudinally on ADC participants in accordance with standard methods. The UDS was implemented at all ADCs on September 1, 2005. Data obtained with the UDS are submitted to the National Alzheimer's Coordinating Center and represent a unique and valuable source of data to support and stimulate collaborative research. PMID- 17132965 TI - MCI is associated with deficits in everyday functioning. AB - The purpose of the present study was to examine the types of impairments in everyday function that accompany mild cognitive impairment (MCI). Data for this study was collected from 434 individuals consecutively evaluated at a university based Alzheimer's Center. A total of 96 participants were diagnosed with MCI, 105 were cognitively normal, and 233 had dementia. Informant ratings of participants' abilities were obtained across different functional domains reflecting everyday abilities related to memory, language, visual spatial abilities, planning, organization, and divided attention. As expected, the demented group was significantly more impaired than the healthy control and MCI groups across all of the functional domains. The MCI group also showed significantly more functional impairment relative to healthy controls in all of the functional domains. Examination of the effect sizes as a measure of the magnitude of functional impairment in the MCI groups relative to controls showed that the greatest degree of impairment occurred within the Everyday Memory domain. The current findings suggest that individuals with MCI demonstrate deficits in a wide range of everyday functions but that the magnitude of these changes is greatest for those functional abilities that rely heavily on memory. PMID- 17132966 TI - 3D mapping of mini-mental state examination performance in clinical and preclinical Alzheimer disease. AB - The Mini-mental State Examination (MMSE) is a brief cognitive screening instrument frequently used to track Alzheimer disease (AD) progression. We investigated the structural neuroimaging correlates of MMSE performance in patients with clinical and preclinical AD. We analyzed structural magnetic resonance imaging data from 29 probable AD and 5 MCI patients who later converted to probable AD using an advanced 3D cortical mapping technique. MMSE scores were entered as covariates in a general linear model that predicted the gray matter density at each cortical surface point. The results were corrected for multiple comparisons by permutation testing. The global permutation-corrected significance for the maps linking gray matter loss and cognitive decline was P=0.005 for the left and P=0.012 for the right hemisphere. Strongest correlations between MMSE score and gray matter integrity were seen in the entorhinal, parahippocampal, precuneus, superior parietal, and subgenual cingulate/orbitofrontal cortices. Significant correlations were also seen bilaterally in the temporal, the middle frontal and the left angular and supramarginal gyri. As a global cognitive measure, MMSE depends on the integrity of widely distributed cortical areas in both brain hemispheres with left-sided predominance. PMID- 17132968 TI - Thyroid function in patients with Alzheimer disease: implications on response to anticholinesterase treatment. AB - Increasing evidence supports an extensive interrelationship between thyroid hormones and the cholinergic system, which is selectively and early affected in Alzheimer disease (AD). The aim of the present study was to explore thyroid function in patients with AD before and after acetylcholinesterase inhibition treatment to possibly identify variances in response. Thyroid function tests were evaluated in 28 AD patients and 24 age and sex-matched controls. Nineteen of the patients were reevaluated after (4 mo) treatment with donepezil. Serum thyrotropin (TSH), triiodothyronine (T3), thyroxine (T4), the free fractions (fT3, fT4) and thyroid autoantibodies were determined using standard methods. All subjects were clinically euthyroid. Patients presented with higher fT4 and anti thyroperoxidase levels, as compared with the controls. Significant reduction in T4, fT3, fT4, and anti-thyroperoxidase levels were observed 4 months after treatment. Responders had higher T4 and fT4, than nonresponders, followed by significant reductions after treatment. The above, within the normal range alterations, may represent a direct effect on hormone release from the thyroid gland and/or increased conversion of T4 to T3 within the brain. Higher T4 and fT4 levels before treatment might predict a favorable response to donepezil treatment. PMID- 17132967 TI - Spanish instrument protocol: new treatment efficacy instruments for Spanish speaking patients in Alzheimer disease clinical trials. AB - OBJECTIVE: To evaluate the feasibility of longitudinal assessment and the psychometric properties of both established and new outcome measures used in clinical trials of patients with dementia in a cohort of Spanish-speaking elders in the United States. METHODS: This is a prospectively collected multicenter study comparing patients with Alzheimer disease (AD) (N=77) and elderly controls (N=17) who are primary Spanish speakers. Spanish-speaking individuals with AD (SSI AD) were selected to represent predefined categories of impairment as determined by a Mini-Mental State Examination score. Controls were selected to approximately match by age and education (SSI C). Subjects were administered a series of Spanish translations of established outcome measures (Mini-Mental State Examination, Clinical Dementia Rating, Geriatric Dementia Scale), and Functional Assessment Staging (FAST)] and new outcome measures developed for United States in clinical trials to assess cognition, function, behavioral disturbance, and clinical global change. Half of the subjects were assessed at 1 and 2 months to evaluate reliability; all subjects were assessed at 6 and 12 months. Comparisons were made between patients and controls and between the Spanish-speaking cohort and a similar English-speaking cohort. RESULTS: The 12-month completion rate was 77%, with a trend toward greater impairment in those with full retention. Both established and new measures demonstrated good internal consistency and test retest reliability in this cohort. All but one measure of cognition demonstrated excellent discriminability between AD subjects and controls. The SSI AD cohort declined significantly on measures of cognition, function, and clinical global change over the 12-month assessment period. The SSI AD and English AD (ESI AD) cohorts declined equivalently on the most common outcomes in clinical trials of AD (delayed recall, clinical global change). Likewise, the most common behavioral changes were also similar in the ESI and SSI groups. However, the annual change was lower in SSI AD than in the ESI AD on several other measures of cognition and function. CONCLUSIONS: These results support the recruitment of Spanish-speaking patients and the use of Spanish language translations for use in the clinical trials for AD. PMID- 17132969 TI - Relationship between the efficacy of rivastigmine and apolipoprotein E (epsilon4) in patients with mild to moderately severe Alzheimer disease. AB - Alzheimer disease is the most common form of dementia in Western countries and the leading cause of disability in the over-65 population. Apolipoprotein E (APOE) is a multifunctional protein implied in lipid metabolism and neurobiology. Polymorphisms of the APOE gene have been associated with a variety of medical disorders, from arteriosclerosis to AD. A high frequency of the APOE epsilon4 allele has been found in patients with AD and they seem to have a higher risk of developing the disease. Various authors have suggested a possible relationship between the efficacy of cholinesterase inhibitors and the presence of the APOE epsilon4 allele. The purpose of the present study was to compare prospectively the efficacy of rivastigmine in patients with mild to moderately severe AD presenting different polymorphisms of the APOE gene on chromosome 19 and to determine if there was a difference in the response to rivastigmine treatment in AD patients with the APOE epsilon4 allele (heterozygous or homozygous) versus patients who had other forms of APOE, such as epsilon2 and epsilon3. This was an open-label, nonrandomized, multicenter study in patients over 50 years of age diagnosed with mild to moderately severe AD. The results of the analysis of this study indicate that the presence of at least one APOE epsilon4 allele does not determine a difference in the response to treatment with rivastigmine. The data indicate that knowledge of the patient's genotype is not necessary for treatment with rivastigmine. It would be interesting in the future to analyze the interaction between these 2 factors using other available anticholinesterase drugs. PMID- 17132970 TI - Cognitive response to memantine in moderate to severe Alzheimer disease patients already receiving donepezil: an exploratory reanalysis. AB - OBJECTIVE: To investigate the cognitive effects of the N-methyl-D-aspartate receptor antagonist, memantine, with a post-hoc exploratory reanalysis of a 24 week randomized, double-blind, placebo-controlled, parallel group clinical trial comparing memantine (20 mg per day) to placebo in patients with moderate to severe Alzheimer disease (AD) receiving treatment with the cholinesterase inhibitor, donepezil. METHODS: The effects of memantine on individual items of the Severe Impairment Battery (SIB), subscale performance, and 3 post-hoc-derived aggregate subscales were investigated. Analyses were based on the intention-to treat population using last observation carried forward and observed cases approaches. The SIB components were assessed at baseline, weeks 4, 8, 12, 18, and 24. RESULTS: The mean change from baseline by visit and at study end point on the SIB showed statistically significant differences between the memantine and placebo groups at all visits beginning at week 8 (last observation carried forward and observed cases). The SIB subscale analysis showed statistically significantly greater effects of memantine than placebo on memory, language, and praxis. When the SIB domains were aggregated using a face valid approach to create 3 higher-order subscales, memantine treatment resulted in statistically significant differences on memory, language, and praxis compared with placebo. CONCLUSIONS: These post-hoc analyses support the beneficial effects of memantine on cognition observed in a previously reported clinical trial. The results presented here suggest an effect of memantine on memory, language, and praxis in patients with moderate to severe AD and support the efficacy of memantine for the treatment of cognitive deficits in AD. PMID- 17132971 TI - Activities of daily living in moderate-to-severe Alzheimer disease: an analysis of the treatment effects of memantine in patients receiving stable donepezil treatment. AB - In moderate-to-severe Alzheimer disease (AD), there are significant losses of activities of daily living (ADL). In a recent prospective, randomized, placebo controlled trial, memantine treatment lessened the overall functional decline in AD patients already on stable donepezil therapy. In this trial, patients (n=404) with Mini-Mental State Examination scores of 5 to 14 receiving stable donepezil treatment were randomized to double-blind treatment with memantine (10 mg b.i.d.; n=203) or placebo (n=201). A primary outcome measure was the 19-item Alzheimer's Disease Cooperative Study--Activities of Daily Living Inventory (ADCS-ADL(19)). To further evaluate the treatment effects of memantine on function, we performed post hoc analyses of ADCS-ADL(19) data from this trial, including ADL items and new subscales derived from factor analysis. Using mixed model analyses, patients receiving memantine had statistically significant less decline in total ADCS ADL(19) scores compared with placebo. An item analysis revealed statistically significant benefits of memantine on grooming, toileting, conversing, watching television, and being left alone. Statistically significant improvements were noted in subscales evaluating higher-level functions and connectedness/autonomy with memantine compared with placebo. These post hoc analyses in moderate-to severe AD patients receiving stable donepezil treatment suggest that memantine may impact overall functional levels, and some of the cognitive processing underlying ADL performance. PMID- 17132972 TI - A phase 2 study of tramiprosate for cerebral amyloid angiopathy. AB - BACKGROUND AND PURPOSE: No treatments have been identified to lower the risk of intracerebral hemorrhage due to cerebral amyloid angiopathy (CAA). A potential approach to prevention is the use of agents that interfere with the pathogenic cascade initiated by the beta-amyloid peptide (Abeta). Tramiprosate (3-amino-1 propanesulfonic acid) is a candidate molecule shown in preclinical studies to reduce CAA in a transgenic mouse model. METHODS: We performed a 5-center phase 2 double-blinded trial to evaluate the safety, tolerability, and pharmacokinetics of tramiprosate in subjects with lobar intracerebral hemorrhage. Twenty-four subjects age > or =55 years with possible or probable CAA were randomized to receive 12 weeks of tramiprosate at 1 of 3 oral doses (50, 100, or 150 mg twice daily). Subjects were followed for clinical adverse effects, laboratory, vital sign, electrocardiogram, cognitive, or functional changes, appearance of new symptomatic or asymptomatic hemorrhages, and pharmacokinetic parameters. RESULTS: Enrolled subjects were younger (mean age 70.8+/-5.4, range 61 to 78) and had more advanced baseline disease (measured by number of previous hemorrhages) than consecutive subjects in a CAA natural history cohort. No concerning safety issues were encountered with treatment. Nausea and vomiting were the most common adverse events and were more frequent at high doses. Nine subjects had new symptomatic or asymptomatic hemorrhages during treatment; all occurred in subjects with advanced baseline disease, with no apparent effect of drug dosing assignment. CONCLUSIONS: These data suggest that tramiprosate can be given safely to subjects with suspected CAA and support future efficacy trials. PMID- 17132973 TI - Efficacy of recreational and occupational activities associated to psychologic support in mild to moderate Alzheimer disease: a multicenter controlled study. AB - We evaluated the efficacy of a stimulation program mainly based on recreational and occupational activities, associated with a brief cycle of support psychotherapy for patients and caregivers, in mild to moderate Alzheimer Disease (AD) associated or not with cerebrovascular lesions. Sixty-seven patients and 31 controls from 2 Italian towns entered the study. The control group was comprised of AD subjects who voluntarily declined to participate in the program for practical reasons. Patients were divided in groups of 4 subjects: treatment was delivered for 6 weeks. Multidimensional efficacy assessment of functional, behavioral, and neuropsychologic aspects was performed. When comparing baseline with posttraining condition, patients displayed a substantial reduction in disruptive behavior, and a tendency to a general reduction of behavioral symptoms compared with controls (Revised Memory and Behavior Problems Checklist--RMBPC- symptoms frequency-total P=0.07; frequency of disruptive behavior P=0.008). This reduction was mirrored by a significant reduction of caregiver reaction to behavioral disturbances (RMBPC caregiver reaction-total P=0.035; reaction to disruptive behavior, P=0.011). At 3 months follow-up, the reduction of caregiver reaction to behavioral symptoms results was confirmed (RMBPC caregiver reaction total P=0.014, caregiver reaction to disruptive behavior P=0.028). No other significant difference was detected. These results partially confirm findings of previous studies, showing that AD patients treated with similar techniques demonstrated an improvement in behavioral disturbances. PMID- 17132974 TI - Health problems and correlates of pain in nursing home residents with advanced dementia. AB - This study describes the health problems and comorbid illnesses of nursing home (NH) residents with advanced dementia (n=123) and identifies correlates of staff identified pain. Study participants were residents of 3 NHs in Maryland, their surrogate decision makers and their physicians. Residents' cognitive function was assessed at study enrollment, and their medical records were reviewed to identify all health problems/illnesses and use of pain medications during the 6 months before their enrollment. The most prevalent health problems were skin problems (95%), nutrition/hydration problems (85%), psychiatric/behavioral problems (85%), gastrointestinal problems (81%), and infections (80%). Sixty-three percent of residents had recognized pain, and 95% of those residents received pain medications. In a multivariate regression analysis, staff-identified pain was associated with aspiration (P=0.008), peripheral vascular disease (P=0.021), musculoskeletal disorders (P=0.032), higher cognitive function (P=0.013), and use of pain medications, including non-opiates (P=0.004) and the combination of opiates and non-opiates (P=0.001). NH residents with advanced dementia experience a complex mixture of multiple chronic and acute comorbidities. These results suggest the need for clinicians in long-term care facilities to be vigilant in assessing and treating pain, particularly as cognitive function declines in those with advanced dementia. PMID- 17132975 TI - Characterization of the kindred of Alois Alzheimer's patient with plaque-only dementia. AB - We describe the kindred of Alois Alzheimer's second published patient (Johann F.) with the brain pathology typical of a subgroup of Alzheimer disease called "plaque-only type." The genealogic records of the kindred extend back to 1670. We constructed a family tree of 1403 individuals and identified 4 living demented members of the Johann F. kindred. The pedigree is consistent with an autosomal dominant trait. The analyses of known dominant dementia genes (APP, PS1, PS2, PRNP, and BRI) failed to reveal mutations in the proband. Further examination of this family might yield new insights into the genetics of Alzheimer disease. PMID- 17132976 TI - Systematic review of the optimal frequency of follow-up in persons with mild dementia who continue to drive. AB - Fitness-to-drive guidelines commonly indicate that persons with mild dementia may be safe to drive but that periodic reevaluation is required. This paper presents the findings of a systematic review of primary evidence regarding the optimal timing of follow-up in persons with mild dementia who continue to drive. A search of Medline, CINAHL, PsychInfo, AARP Ageline, and Sociofile from 1984 to 2005 was performed. No published studies focus primarily on the timing of follow-up of drivers with mild dementia. Three studies present longitudinal data that the authors reference when recommending periodicity of follow-up. This study identifies a concerning research gap in the field of dementia and driving. To provide better evidence to guide recommendations for periodicity of follow-up, 3 recommendations are proposed: (1) that prospective cohort driving research be undertaken to follow patients with mild dementia who continue to drive, (2) that data from such longitudinal research be presented as survival analyses, and (3) that existing research on the progression rates of Alzheimer disease be employed as a default until the first 2 recommendations are realized. PMID- 17132977 TI - Insulin resistance syndrome and Alzheimer disease: pathophysiologic mechanisms and therapeutic implications. AB - Insulin plays a key role in cognition and other aspects of normal brain function. Insulin resistance induces chronic peripheral insulin elevations, reduces insulin activity, and reduces brain insulin levels. The insulin resistance syndrome and associated conditions such as type 2 diabetes mellitus and hypertension, are associated with age-related memory impairment and Alzheimer disease. Our work has focused on potential mechanisms through which this association is forged, including the effects of peripheral hyperinsulinemia on memory, inflammation, and regulation of the beta-amyloid peptide. We have shown that raising plasma insulin to levels that characterize patients with insulin resistance invokes synchronous increases in levels of beta-amyloid and inflammatory agents. These convergent effects may impair memory and induce AD pathology. Therapeutic strategies focused on preventing or correcting insulin abnormalities may thus benefit adults with age-related memory impairment and AD. PMID- 17132978 TI - Cognitive change in Parkinson disease. AB - Parkinson disease (PD) is the most common neurodegenerative movement disorder, affecting 1 in 100 individuals over the age of 60. Dementia in the setting of PD (PDD) may be among the most debilitating symptoms associated with disease progression. Estimates of cognitive decline and dementia in PD suggest that up to 14% per year of patients over age 65 with PD will develop some cognitive impairment. Unfortunately, PDD is not well characterized and the relationship of PDD to Alzheimer disease remains unclear. PDD has been proposed as part of a spectrum with dementia with Lewy bodies, and PDD and dementia with Lewy bodies frequently coexist with Alzheimer disease. It is uncertain, however, whether there is a meaningful distinction between the different disorders. It has also been difficult to gain understanding of the interaction of motor and non-motor symptoms that affect quality of life in PD and confound cognitive and psychomotor performance. This review will examine the clinical, cognitive, neuropsychiatric features of cognitive deficits associated with PD, discuss their pathologic basis and propose avenues for future research. PMID- 17132979 TI - Third Asia-pacific regional meeting of the international working group on harmonization of dementia drug guidelines: meeting report summary. AB - The International Working Group on Harmonization of Dementia Drug Guidelines has held the third Asia-Pacific Chapter meeting in 2004. The committee concludes that the Neuropsychiatric Inventory and the Clinical Dementia Rating are culturally acceptable assessment tools. The epidemiology studies of mild cognitive impairment and dementia in Asia were discussed at the meeting. The committee encouraged a collaborative research in the region in the future. PMID- 17132980 TI - Workshop reports from the third Asia-pacific regional meeting of the international working group on harmonization of dementia drug guidelines. PMID- 17132981 TI - Neuropsychiatric inventory workshop: behavioral and psychologic symptoms of dementia in Asia. AB - The Neuropsychiatric Inventory (NPI) was introduced in 1994 and has since become a standard instrument for clinical trials and other types of behavioral research in dementing disorders. Its reliability and validity have been confirmed. The NPI was the subject of a workshop in Asia in conjunction with the International Workgroup on Dementia Drug Guidelines (IWG). Investigators using the NPI from 4 Asian areas--Taiwan, Hong Kong, Japan, Thailand--presented conclusions from their research. A high prevalence of behavioral disturbances across Asian countries was found and the rates are similar to those observed in Western countries. Apathy is more difficult to detect and characterize in Asian populations. Neurobiologic studies show an excess of some serotonin receptor gene polymorphisms in patients without behavioral disturbances and positron emission tomography reveals reductions in frontal lobe metabolism in patients manifesting depression as measured by the NPI. Studies in Thailand show relationships among verbal fluency, activities of daily living, and neuropsychiatric symptoms particularly agitation, apathy, and disinhibition. This suggests a triad of symptoms of behavioral abnormalities, executive dysfunction, and abnormalities of activities of daily living that impugn frontal lobe function. The NPI is a reliable and useful instrument to characterize behavioral changes in Asian and Western populations. PMID- 17132982 TI - Clinical dementia rating workshop: the Asian experience. PMID- 17132983 TI - Estrogen receptor beta gene (ESRbeta) 3'-UTR variants in Alzheimer disease. PMID- 17132984 TI - New insight into Alzheimer disease: demonstration of fibrin(ogen)-serum albumin insoluble deposits in brain tissue. PMID- 17132985 TI - Recurrence of upper gastrointestinal bleeding after donepezil administration. PMID- 17132999 TI - Pay for performance. PMID- 17133001 TI - On the cover. PMID- 17133003 TI - Alzheimer disease and genetics: anticipating the questions. AB - Three genes with autosomal dominant mutations have been identified that may lead to Alzheimer symptoms in carriers before they reach age 60. Genetic tests exist for Alzheimer disease, but they are considered useful only for the small number of families with a history of early-onset illness. As researchers continue to uncover evidence of genetic links to Alzheimer disease, nurses can expect to field questions from family members about genetic testing. The article presents a variety of questions nurses may be asked, as well as possible answers. PMID- 17133005 TI - High school students take classes in a nursing home. PMID- 17133007 TI - The local view abroad. PMID- 17133008 TI - International recruitment. PMID- 17133009 TI - Methamphetamine abuse. PMID- 17133010 TI - Silver-containing dressings and the need for evidence. AB - The field of wound care has been inundated with "active" dressings--those that deliver biologically active substances to the wound site--and much attention has been given to those containing silver. Silver is a broad-spectrum agent: it is bactericidal to a large number of Gram-positive and Gram-negative microorganisms, many aerobes and anaerobes, and several antibiotic-resistant strains. More than 10 dressings containing pure silver are currently available; all claim to enhance wound healing through the antimicrobial activity of silver. But research findings on silver's antimicrobial efficacy are often inconclusive or contradictory. How, therefore, can clinicians decide which dressing is most appropriate for each patient? A grading system for reviewing the literature is presented. PMID- 17133012 TI - Erectile dysfunction and its discontents. PMID- 17133017 TI - HIV risk reduction among African American teenage girls. PMID- 17133021 TI - Postmortem ophthalmologic examination by endoscopy. AB - Abnormal intraocular findings detected during forensic autopsies have generally been ignored. In this study, we assessed the usefulness of postmortem ophthalmologic examination in 46 cases using an endoscope system with a tip diameter of 0.9 mm. This procedure was performed using a portable system, and only 1 assistant was needed. Furthermore, the procedure required only about 5 minutes for both eyes. Abnormal intraocular findings were seen in 22 (47.8%) of 46 cases. Fundus hemorrhages were observed in 4, and papilledema was observed in 3 of 5 cases of death due to head trauma; besides, it was possible to estimate the interval between head trauma and death based on intraocular findings. Seven of 16 cases of death from asphyxia presented abnormal intraocular findings, including retinal splinter hemorrhages. Fundus hemorrhages were observed in 2 of 3 cases of death by drowning, which, to our knowledge, had never been reported to date.Thus, this procedure proved easy to perform, and it may be useful to detect significant abnormal intraocular findings related to the presumed cause of death during autopsies. PMID- 17133022 TI - Mechanisms of aortic blunt rupture in fatally injured front-seat passengers in frontal car collisions: an autopsy study. AB - We tried to explain the mechanisms of the aortic blunt ruptures in fatally injured drivers and front passengers, unrestrained by seatbelts, by analyzing the frequencies of both aortic ruptures and concomitant injuries to 12 organs and body regions. The sample consisted of 393 subjects: 251 drivers and 142 front passengers (325 male and 68 female passengers, the mean age 41.0 +/- 15.5). The total number of the complete blunt aortic ruptures in the sample was 116 (80 in the drivers and 36 in the front passengers). The weakest part of the aorta seems to be the isthmus (47 isthmus ruptures in the drivers and 27 in the front passengers). The statistically significant concomitant injured organs and body regions with the aortic ruptures were the liver, the sternum, and the diaphragm in the car drivers and the head and the neck in the front passengers. According to these results, the mechanisms of thoracic aorta rupture are different for fatally injured drivers and front passengers. For car drivers, they are associated and simultaneous with both thoracic and abdominal compression due to deceleration of the body at the moment when the driver's body slides forward and flexes across and against the steering wheel. For the front passengers, the mechanism is the caudorostral hyperextension of the thoracic aorta at the moment when the body is stopped by a dashboard, but the head continues forward with great velocity: the carotid vessels pull the aortic arch forward at the same time as the intercostal arteries fix the thoracic part of the aorta and pull it downwards. PMID- 17133023 TI - Image analysis as an adjunct to sodium rhodizonate test in the evaluation of gunshot residues: an experimental study. AB - It is important to investigate the clothing, as well as the body, to determine the range of fire of entry wounds in firearm injuries. Clothing can affect the amount of gunshot residues (GSR) reaching the body and their distribution. The amount and distribution of the GSR also vary according to the distance between the firearm and the target. Sodium rhodizonate test provides valuable data when clothing is available for examination. In the absence of clothing, light microscopic examinations may add additional information regarding the range of fire. In this study, a sodium rhodizonate test was done on 80 garment samples containing the bullet entrance. The 80 calfskin samples were processed histologically and were stained using Alizarin Red S. These were also evaluated with computer-assisted image analysis. Gross residues were seen on military camouflage clothing in samples from < or = 45-cm group. White flannel undershirts under the military camouflage contained rhodizonate-positive particles only around the contact wounds. With image analysis, however, the residues could be detected also in the skin samples of the 2.5-cm- and 5-cm-range groups. We suggest that the image analysis can be combined with other techniques and it can provide valuable data in the determination of entry wounds and also in the estimation of firing distance. PMID- 17133024 TI - Death caused by myocarditis in Wayne County, Michigan: a 9-year retrospective study. AB - This paper reviews 72 cases of death caused by myocarditis between the years 1996 and 2004, autopsied at the Office of the Wayne County Medical Examiner in Michigan. Myocarditis as a cause of sudden and unexpected death represented 1.3% of all natural deaths in Wayne County during said period. The year 1999 contained the highest number of deaths of this cause (18), where the average number of myocarditis deaths was 8 per year for this 9-year span. In this study, each case was reviewed based on information gathered from investigative, autopsy, and toxicology reports. Significantly, 58% of these cases were male, and 63.4% were African American. Myocarditis caused death in every age group between 7 months and 67 years, but adults between the ages of 19 and 67 were most significantly affected (75%). Flu and/or cold were the most common symptoms experienced in the days directly proceeding death (28%), followed by shortness of breath (17%) and sudden collapse (15%). Sixty-nine percent of these 72 cases were pronounced dead after ACLS (advanced cardiac life support) protocol by emergency medical services or hospital attendants. Cardiomegaly was observed in 24 cases of adults aged 19 or older (54%), and flabby/soft myocardial tissue was observed grossly in 16% of all 72 cases. PMID- 17133025 TI - The deadly broomstick: an unusual missile injury to the neck. AB - A 51-year-old man was struck by the tip of a broomstick weighing 1000 g at the left side of the neck, upon which he collapsed. Intense but delayed cardiopulmonary resuscitation restored the circulation roughly 30 minutes after the incident. Upon admittance to a nearby hospital, an extensive hypoxic cerebral damage was diagnosed. Death due to the severe cerebral damage occurred 5 hours after the incident. An autopsy demonstrated a severe subcutaneous traumatization of the left side of the neck, with a hemorrhage compressing the carotid bifurcation. A prolonged excitation due to this ongoing compression of the baroreceptors in the carotid sinus was assumed to have led to a cardiac arrest. In this case report, the authors discuss the underlying pathophysiology of this potentially lethal and rare reflexogenic incident also known as the Hering reflex and discuss possible therapeutic measures. PMID- 17133026 TI - Dismemberment: a review of the literature and description of 3 cases. AB - The authors describe 3 cases of dismemberment. Numerous methods of hiding a body and thus erasing proof of murder have been devised. Those most frequently described in the literature include: burial of the body in an unusual or impervious place such as a wood, grotto or mountain; charring the body by wrapping it in a tire, for example, to fuel the fire until all traces of the crime have been erased and identification of the victim is difficult; and dropping the weighted-down body in the open sea or in a well in the hope that it will never be found. Dismemberment is in reality a relatively rare method whereby, after killing the victim, the murderer uses a very sharp cutting weapon (a saw, axe, etc.) to sever the limbs and cut the body into small pieces. The operation is generally carried out immediately after the crime, although more rarely a long time may pass between the 2 events. PMID- 17133027 TI - Pink teeth in a series of bodies recovered from a single shipwreck. AB - Pink teeth have most often been observed in victims of drowning but have also been reported in subjects who died suddenly and unnaturally. There is general agreement that there is no obvious connection between the occurrence of pink teeth and the cause of death, but the condition of the surroundings (especially humidity) must certainly play an important role in the development of the pink tooth phenomenon. The frequency and distribution of postmortem pink coloration of the teeth have been studied among a representative sample of 52 cadavers. All the bodies were victims of a single shipwreck that occurred on March 13, 1997, in the middle of the Otranto Canal (Mediterranean Sea). The bodies were recovered from the seawater after approximately 7 months. A distinct pink coloration of the teeth was found in only 18 cadavers (13 females and 5 males) of ages ranging between 13 and 60 years. The phenomenon was more pronounced in younger individuals due to age-related changes of the root canal, less penetrable by the pigment responsible for the postmortem pink staining. By histochemical methods and autofluorescence, hemoglobin and its derivatives have been identified as the most likely pigments responsible for this postmortem process that can be considered analogous to postmortem lividity. These data are consistent with previous reports on pink teeth, indicating that the diffusion of the blood in the pulp into the dentinal tubules causes the red discoloration of the teeth. Based on the results, the pigmentation is more prominent on the teeth with single roots rather than in the posterior teeth with multiple roots. PMID- 17133028 TI - Sudden infant death due to neurofibromatosis type I. AB - Sudden infant death syndrome (SIDS) is the unexpected death of an infant under the age of 1 year, where a complete autopsy, including scene investigation, fails to reveal a cause of death. Although the frequency of SIDS has decreased almost 50% over the past 10 years, it remains the leading cause of death in infants aged 1 to 6 months. SIDS is a diagnosis of exclusion and requires the elimination of a wide range of possible causes, including asphyxia, poisoning, abuse, occult heart disease, and other natural disease processes. In this report, we describe the case of an infant death initially suspected to be a SIDS death in which autopsy revealed an optic pathway glioma (optic glioma or hypothalamic glioma) and other stigmata of neurofibromatosis type I. PMID- 17133029 TI - Chronic hydrocephalus and alcohol abuse in a young male suicide. AB - The paper describes a case of suicide in a young man affected by compensated chronic hydrocephalus who was subject to alcohol abuse. The case was studied by means of a complex set of analyses, including circumstantial and clinical data, anatomohistopathological findings, and chemicotoxicologic tests. What clearly emerges in the case is the importance of a continuing neuropsychological follow up in patients with shunted hydrocephalus. The forensic interest in the case is due to the peculiar autopsy findings discussed in relation to the possible causes of sudden death in subjects with hydrocephalus. PMID- 17133030 TI - Heat stroke deaths caused by electric blankets: case report and review of the literature. AB - Heat stroke is the most serious and potentially life-threatening condition of the heat-related illnesses. Heat stroke deaths caused by electric blanket are rarely reported. In this paper, we report 2 cases of fatal heat stroke caused by overheating from electric blankets in winter. One was a 41-year-old man who was found unresponsive in bed on an electric blanket. His wife shared the same bed with him and was found unconscious. The wife's axillary temperature was 40 degrees C (104 degrees C) when she was admitted to the hospital. She fully recovered after medical treatment. The husband was pronounced dead at scene, with rectal temperature at 41.2 degrees C (106.2 degrees C). The other was a 13-year old girl who was found dead in bed on an electric blanket, with rectal temperature at 41 degrees C (105.8 degrees F). The literature is reviewed, and the pertinent findings, including scene investigations, postmortem examination, the risk and mechanism of fatal heat stroke caused by using electric blanket, are discussed. PMID- 17133031 TI - Blood levels of 3-methylfentanyl in 3 fatal poisoning cases. AB - Three fatal poisoning cases due to 3-methylfentanyl are described. In each case, the death was accidental and occurred after injection of the opioid combined with amphetamine, heroin, or other drugs. The victims' ages, ranging from 30 to 41 years, were higher than those typically found in heroin poisonings in Finland. The blood concentrations of cis-3-methylfentanyl, measured here for the first time by a specific tandem mass spectrometric method, ranged from 0.3 to 0.9 microg/L (mean 0.5 microg/L). These values are significantly lower than the levels reported for alpha-methylfentanyl and fentanyl in fatal poisonings. Repeated seizures of fentanyl and its analogs have been reported in Europe close to the Russian border. PMID- 17133032 TI - Fatalities due to intoxicated arrestees jumping out of moving police vehicles. AB - Fatalities resulting from emergency vehicle crashes are relatively rare. Mortality, particularly in the case of unbelted occupants, is relatively high. Data on drunken (handcuffed) arrestees jumping out of a moving police vehicle or the circumstances of such events have not been published. Two cases of such fatalities are described in this paper. Since these cases should be considered as "death in custody," may give rise to significant covering and speculation in the media, and may raise liability questions, they require an in-depth medicolegal investigation, including investigation of the crime scene, complete medicolegal autopsy/toxicology, and reconstruction of the event in the presence of an experienced forensic pathologist. From the 2 cases described in this paper, it appears that mechanical malfunctioning of the locking device of the door of the police vehicle and lack of controlling the actual lock-tied closing of the vehicle door (instead just assuming that it happened) may precipitate such cases. Auditory control by the police officers of the arrestee tampering with or manipulating the car door may be hampered by the background noise of the police vehicle, the on-board radio, and the use of a siren. Regular visual control and adequate control of the actual locking of the vehicle door are of paramount importance to prevent such mishaps. Reconstruction of the event in the presence of the forensic experts is mandatory to test the different hypotheses of the fatal event. PMID- 17133033 TI - TB-related sudden death (TBRSD) due to myocarditis complicating miliary TB: a case report and review of the literature. AB - TB-related sudden death (TBRSD) is rarely reported in the literature and in the majority of cases is due to bronchopneumonia and hemoptysis. Cardiac complications of tuberculosis causing sudden death can take many forms and are rarer still, with only a handful of cases reported. We describe a case of a previously fit and healthy 20-year-old Asian female who, after returning from a holiday in India, collapsed while getting off a bus. At postmortem, the only macroscopic finding of note was a localized area of fibrosis on the anterior wall of the left ventricle. Microscopic examination of this area showed Langhans giant cells; noncaseating epithelioid granulomas and acid-fast bacilli were demonstrated on Ziehl Nielsen staining. In addition, the lungs, liver, and kidneys contained multiple noncaseating granulomas. The case serves to highlight the protean nature in the presentation of this disease and the importance of postmortem histology in autopsy work. PMID- 17133034 TI - Avulsion of the distal tibial shaft in aircraft crashes: a pathological feature of extreme decelerative injury. AB - A pilot and his passenger were killed on impact with the ground when their light aircraft crashed. Both deaths were caused by extensive injuries involving severe craniocerebral, skeletal, soft tissue, and organ trauma. In both victims, the legs were shortened, with stripping of muscle and soft tissues from the shafts of the lower limb long bones. In addition, fragments of distal tibial shaft had been forced through the soles of the victims' shoes. This sign indicated a fall from height and showed that the direction of the decelerative forces had been along the axis of the legs and that the force of impact was severe enough to cause fracturing of the lower limb bones, with subsequent extrusion of bone fragments downwards through the shoes. When present, this observation represents another feature at autopsy that can add to the understanding of the circumstances of a fatal air crash and the position of the victims immediately prior to impact. PMID- 17133035 TI - The role of beta-amyloid precursor protein (beta-APP) staining in the neuropathologic evaluation of sudden infant death and in the initiation of clinical investigations of subsequent siblings. AB - This report highlights the importance of undertaking immunohistochemical staining of the brains of infants who die unexpectedly, as it may not only assist with the evaluation of the cause of death in an individual infant but may also help with the clinical management of subsequent siblings. A 5-month-old male infant who died suddenly was found to have diffuse beta-amyloid precursor protein (beta-APP) staining in the brain, with no unusual features in his history, death scene examination, routine autopsy dissection, and ancillary tests to suggest any definite cause of death. Due to the beta-APP staining, the possibility of previous episodes of occult trauma, apparent life threatening events (ALTEs), and accidental or inflicted suffocation was raised in the autopsy report. As detailed analyses and investigations provided no supportive evidence for trauma or inflicted injury, hypoxia was clinically considered the most likely cause. Because of these concerns, sleeping oxygen saturation levels were monitored following the birth of a subsequent sibling who had normal APGAR scores and no evidence of any health problems. Oxygen desaturation to 70% occurred in association with a color change while on the postnatal ward, and a subsequent polysomnogram showed multiple episodic significant desaturations to around 80% in association with central apnea. Other testing was unremarkable. These cases demonstrate that beta-APP staining of the brain may not only provide clues as to possible mechanisms of death in pediatric forensic cases but may indicate a need for careful clinical evaluation of subsequent siblings for possible central apnea requiring oxygen therapy. PMID- 17133036 TI - Amended forensic autopsy legal procedures in Turkey during integration with the European Union (EU). AB - European Union (EU) is expanding its territories, and Turkey has been making adaptive changes in many aspects for an integration with EU. In this regard, amendments on forensic autopsy procedures have been effective from June 1, 2005. This article delineates these changes, emphasizing the differences between the previous and the current procedures. PMID- 17133037 TI - Highly sensitive HLA-DNA typing from formalin-fixed and paraffin-embedded tissue samples. AB - Nucleotide sequences have been determined for more than 1700 different alleles at the core of the human leukocyte antigen (HLA) system. The highly polymorphic character of these genes affects adaptive immune response and is also useful for forensic applications. HLA typing from formalin-fixed and paraffin-embedded tissue provides abundant useful information for both clinical settings and forensic investigations. This study, which investigated the potential use of DNA from formalin-fixed and paraffin-embedded tissue samples in an HLA PCR sequence specific primer and probe (SPP) system, showed that tissue fixed in formalin for less than 3 days and embedded in paraffin can serve as a useful source of DNA for PCR-SPP typing kits. PMID- 17133038 TI - Mind your manners: quality of manner of death certification among medical examiners and coroners in Taiwan. AB - To assess the quality of manner of death (MOD) certification among medical examiners/coroners (ME/Cs) in Taiwan, death certificates issued in 2002 for which the final MOD was suicide or undetermined were extracted for analysis. Indicators of the quality of MOD certification included (1) MOD not given by the ME/Cs; (2) MOD assigned by the ME/Cs was changed by the coder; (3) ratio between undetermined and suicide deaths (U/S ratio). There were 450 death certificates for which the ME/Cs did not assign the MOD in the original certificate. Three fifths (285/450) of them were issued by 4 ME/Cs. The same 4 ME/Cs also had extremely high U/S ratios (1.25-1.84) compared with the average (0.31). The overall quality of MOD certification among ME/Cs in Taiwan was fair; only a small number of ME/Cs had poor quality in MOD certification. The high U/S ratio among the 4 ME/Cs would certainly affect the suicide mortality rates of the counties the 4 ME/Cs were in charge of. Actions should be taken to improve the certification quality of these 4 ME/Cs. PMID- 17133039 TI - Evaluation of certifier practices regarding alcohol-related deaths: Fulton County Medical Examiner's Center, Atlanta, Georgia, 2004. AB - CONTEXT: Alcohol can contribute to various manners of death by acute intoxication that places a person at risk for fatal injury, acute fatal alcohol poisoning, or the various fatal complications of chronic abuse with or without superimposed acute intoxication. The reporting of alcohol use on the death certificate may vary with office policy or procedure, certifier judgment, and the timing of information received during investigation. OBJECTIVE: To determine the number of deaths including mention of alcohol use in the investigative case file, the number of death certificates on which alcohol use is reported, the number of discrepancies between the 2, and the possible reasons for observed discrepancies. DESIGN, SETTING, AND PARTICIPANTS: Retrospective case review of all deaths where alcohol use was mentioned in the investigative case file and/or on the death certificate for deaths investigated by the Fulton County Medical Examiner in Atlanta, Georgia, during a 1-year period between January 1, 2004, and December 31, 2004. MAIN OUTCOME MEASURES: Percentage of deaths with alcohol use reported on the death certificate, tabulation of where and how alcohol use is reported on the death certificate, and tabulation of the differences between the investigative case file and death certificate regarding alcohol's possible role in causing death. RESULTS: Among the 1324 deaths certified by the office, 105 (8%) had alcohol use reported on the death certificate. The majority (67%) of these cases were natural deaths. Sixty-nine (5%) deaths had mention of alcohol use in the investigative case notes but did not include it on the death certificate. Twenty-five (2%) deaths had mention of alcohol on the death certificate but did not have mention of it in the investigative case file based on our search criteria. However, subsequent review of additional case follow-up information disclosed a history of alcohol use or acute intoxication in each case. CONCLUSIONS: The data show that more natural deaths are considered to be directly caused by alcohol than other manners of death. For the unnatural manners of death (excluding acute alcohol poisoning), alcohol use is often viewed by medical examiners as an incidental, associated finding or risk factor surrounding the circumstances of death rather than being an actual cause of death. In such cases, alcohol use is often omitted from the death certificate. For deaths directly caused by alcohol, the proportion of cases involving possible underreporting or overreporting of alcohol involvement was relatively small and usually involved the omission of chronic alcohol use from the death certificate. Researchers need to be aware of potential limitations of death certificate data for studying alcohol-related deaths. PMID- 17133040 TI - Postmortem urine immunoassay showing false-positive phencyclidine reactivity in a case of fatal tramadol overdose. AB - This is a report of postmortem false-positive reactivity using an enzyme multiplied urine phencyclidine (PCP) immunoassay (EMIT II+) due to a single-agent fatal tramadol overdose. An autopsy of a 42-year-old male who died alone at home revealed no identifiable lethal anatomic abnormalities, thus leading to toxicologic analysis. Femoral blood was obtained for drug testing by high performance liquid chromatography (HPLC) and showed a tramadol level of 14.0 mg/L, 2 orders of magnitude greater than the therapeutic range (0.1 to 0.3 mg/L). Urine was also obtained and EMIT II+ immunoassay revealed positivity for PCP at 88 mAU/min. However, confirmatory testing by HPLC failed to identify PCP in either the urine or serum. To verify the suspicion that this was a false-positive PCP result, stock solutions of tramadol and its major metabolite (O desmethyltramadol) at concentrations of 100 mg/L in 10% methanol/H2O were compared with a blank solution (10% methanol/H2O) for EMIT II+ PCP reactivity and demonstrated reactivities of 44 mAU/min and 27 mAU/min, respectively. While these individual results were below the cutoff reactivity for a positive EMIT II+ PCP result (ca. 85 mAU/min), they were much more reactive than the blank calibrator (set at 0 mAU/min). Therefore, we conclude that the immunoreactivity of tramadol and its metabolites in aggregate is responsible for the PCP immunoassay interference and false-positive result. PMID- 17133042 TI - Case analysis of brain-injured admittedly shaken infants: fifty-four cases, 1969 2001: a reply. PMID- 17133043 TI - Pseudophakic corneal edema: A review of mechanisms and treatments. AB - PURPOSE: To review the pathological mechanisms and treatments for pseudophakic corneal edema (PCE), one of the most common indications for penetrating keratoplasty. METHODS: The literature was examined for the molecular biology associated with PCE and for the surgical and medical treatments for this disorder. RESULTS: The incidence of PCE has recently been decreasing because of improved surgical instrumentation, including improvements in intraocular lens designs that cause less trauma to the corneal endothelium. Extracellular matrix and growth factor abnormalities occur in PCE corneas and recently, the role of aquaporins, which are involved in the regulation of fluid movement across cells, has been investigated. CONCLUSIONS: Although newer treatment options have been suggested, penetrating keratoplasty still remains the most definitive treatment and has the highest success rate. PMID- 17133044 TI - Risk factors for the first episode of corneal graft rejection in keratoconus. AB - PURPOSE: To evaluate the relationship between topical corticosteroids and other variables and the risk for rejection after penetrating keratoplasty for keratoconus. METHODS: The records of all keratoconus patients who, after their first penetrating keratoplasty in that eye, experienced a first episode of corneal graft rejection during a specific 3-year period were retrospectively reviewed in a case-control fashion. Twenty-three cases were identified, and they were matched with 3 controls each, for a total of 69 controls and 92 total patients. Multiple variables including steroid potency, recent steroid tapering, and length of time on the current level of steroids were analyzed to see whether there were any significant relationships between postoperative changes in steroid management and rejection. In addition, other variables such as graft size, suture technique, recent suture removal, suture status at the time of the rejection episode, and prior grafting in the fellow eye were examined to determine if any of these factors were associated with a higher risk of graft rejection. RESULTS: Most of the proposed risk factors, including steroid dose and tapering, differing suturing techniques, loose and/or broken sutures at the time of rejection, percentage of sutures remaining at the time of rejection, and prior grafting in the fellow eye, did not correlate with the risk of rejection. Only graft size had a correlation, with host trephination size > or = 8.25 mm having a nearly sixfold increased risk of rejection (P = 0.015). Most patients (70%) were diagnosed with rejection at a scheduled office visit rather than at an emergency visit, and correspondingly, nearly one half (43%) had no symptoms when rejection was identified. There was no significant difference in final best-corrected visual acuities between the cases and controls, and 91% of the corneas that underwent rejection did not progress to graft failure, remaining centrally clear at most recent follow-up. CONCLUSION: In this study, the most important risk factor for rejection after corneal transplantation for keratoconus was the size of the graft. Physician detection of rejection is paramount, because a graft rejection episode is more often diagnosed at a scheduled office visit than at an emergency visit. Fortunately, progression to graft failure can usually be prevented if treatment is started promptly and intensively. PMID- 17133045 TI - Effect of antibacterial honey on the ocular flora in tear deficiency and meibomian gland disease. AB - PURPOSE: To assess for differences in the ocular flora of patients with dry eye caused by tear deficiency and/or meibomian gland disease and to assess the effect of antibacterial honey on the ocular flora in these forms of dry eye. METHODS: In this prospective, open-label pilot study, bacteria isolated from the eyelid margin and conjunctiva were identified and quantified before and at 1 and 3 months after initiation of treatment with topical application of antibacterial honey 3 times daily. Subjects had non-Sjogren tear deficiency (n = 20), Sjogren syndrome tear deficiency (n = 11), meibomian gland disease (n = 15), and non Sjogren tear deficiency with meibomian gland disease (n = 20), and there were 18 non-dry eye subjects. RESULTS: The total colony-forming units (CFUs) isolated from each of the dry eye subgroups before antibacterial honey use was significantly greater than the total CFU isolated from the non-dry eye group. Antibacterial honey use significantly reduced total CFUs for the eyelids and the conjunctiva of dry eye subjects from baseline at month 1 (eyelids: P = 0.0177, conjunctiva: P = 0.0022) and month 3 (eyelids: P < 0.0001, conjunctiva: P < 0.0001). At month 3, there were reductions in total CFUs for all dry eye subgroups such that the CFUs were not significantly different from those of the non-dry eye group. CONCLUSION: From these results, there is sufficient preliminary data to warrant further study of the effects of antibacterial honey in chronic ocular surface diseases. PMID- 17133046 TI - Simultaneous versus sequential penetrating keratoplasty and cataract surgery. AB - PURPOSE: To compare the surgical outcomes of simultaneous penetrating keratoplasty and cataract surgery with those of sequential surgery. METHODS: Thirty-nine eyes of 39 patients scheduled for simultaneous keratoplasty and cataract surgery and 23 eyes of 23 patients scheduled for sequential keratoplasty and secondary phacoemulsification surgery were recruited. Refractive error, regular and irregular corneal astigmatism determined by Fourier analysis, and endothelial cell loss were studied at 1 week and 3, 6, and 12 months after combined surgery in the simultaneous surgery group or after subsequent phacoemulsification surgery in the sequential surgery group. RESULTS: At 3 and more months after surgery, mean refractive error was significantly greater in the simultaneous surgery group than in the sequential surgery group, although no difference was seen at 1 week. The refractive error at 12 months was within 2 D of that targeted in 15 eyes (39%) in the simultaneous surgery group and within 2 D in 16 eyes (70%) in the sequential surgery group; the incidence was significantly greater in the sequential group (P = 0.0344). The regular and irregular astigmatism was not significantly different between the groups at 3 and more months after surgery. No significant difference was also found in the percentage of endothelial cell loss between the groups. CONCLUSION: Although corneal astigmatism and endothelial cell loss were not different, refractive error from target refraction was greater after simultaneous keratoplasty and cataract surgery than after sequential surgery, indicating a better outcome after sequential surgery than after simultaneous surgery. PMID- 17133047 TI - Long-term graft survival in patients with Down syndrome after penetrating keratoplasty. AB - PURPOSE: To determine graft survival and long-term visual outcome after penetrating keratoplasty (PK) for keratoconus in patients with Down syndrome. METHODS: The records of all patients with Down syndrome who received PK by the same provider were reviewed. A retrospective analysis was performed to determine long-term graft survival, incidence of graft failure, and complication rate. RESULTS: Twenty-one PKs were performed on 18 eyes of 13 patients with Down syndrome with keratoconus. Three repeat PKs were performed for secondary graft failure. All 18 eyes had clear grafts at the most recent examination. Follow-up ranged from 4 to 88 months, with a mean of 34.9 months. The average age of patients was 42 years, with a range of 20 to 63 years. Preoperative visual acuity ranged from 20/160 to count fingers. Postoperatively, visual acuity was objectively measurable in 12 eyes of 8 patients and ranged from 20/30 to 20/200, with a mean of 20/60. Broken sutures and difficulties with unsedated suture removal complicated postoperative care in some patients. CONCLUSION: Clear grafts and improvements in visual acuity can be obtained after PK in patients with Down syndrome, but consideration must be given to careful postoperative care by health care providers and home support personnel. PMID- 17133048 TI - Comparison of the effect of three suturing techniques on postkeratoplasty astigmatism in keratoconus. AB - PURPOSE: To compare the effect of 3 common suturing techniques on postkeratoplasty astigmatism and final best corrected visual acuity (BCVA) in patients with keratoconus. METHODS: In this randomized clinical trial, 103 eyes of 103 patients with advanced keratoconus, who were contact lens intolerant, or with contact lens-corrected visual acuity less than 20/80, underwent penetrating keratoplasty with 3 suturing techniques: interrupted (IR), single running (SR), and combined interrupted and running (CIR). Postkeratoplasty astigmatism and BCVA were evaluated during regular examinations 1.5, 3, 6, and 12 months postoperatively and 2 months after complete suture removal. Suture adjustment and selective suture removal were performed 2 to 6 weeks and after 3 months in eyes with more than 4 D of corneal astigmatism in the SR and IR/CIR groups, respectively. RESULTS: Of 87 patients who completed follow-up, 26 eyes underwent PK with interrupted suturing technique (IR), 26 eyes had single running sutures (SR), and in 35 eyes, the suturing technique was combined (interrupted + running sutures; CIR). Mean age was 27.2 +/- 8.4, 28.9 +/- 8.7, and 30.3 +/- 8.7 years, and postoperative astigmatism 1.5 months after surgery was 3.77 +/- 1.68, 5.48 +/ 2.09, and 4.10 +/- 1.79 D in the 3 groups, respectively (P = 0.015). However, 2 months after complete suture removal, final postoperative astigmatism was 3.83 +/ 1.65, 3.37 +/- 1.9, and 3.88 +/- 2.79 D (P = 0.851) and BCVA (log MAR) was 0.08 +/- 0.14, 0.13 +/- 0.23, and 0.09 +/- 0.16, respectively (P = 0.53). Immunologic endothelial rejection reactions were seen in 5 eyes (19.2%) in the IR group, 3 eyes (11.72%) in the SR group, and 6 eyes (17.64%) in the CIR group (P = 0.44). There was no case of graft failure during the follow-up period. CONCLUSION: Postkeratoplasty astigmatism and BCVA are comparable with the 3 common suturing techniques (IR, SR, and CIR) in patients with keratoconus, provided that regular postoperative examinations and topography-guided suture adjustment and/or removal are performed. PMID- 17133049 TI - AlphaCor: Clinical outcomes. AB - PURPOSE: To study the outcomes of AlphaCor implantation. METHODS: : The AlphaCor artificial cornea is indicated for corneal blindness not treatable by donor grafting. Prospective preoperative and follow-up data were collected. Data were evaluated using SPSS for statistical analysis of outcomes, trends, and associations. RESULTS: This report includes data returned through February 28, 2006, for all 322 devices implanted, with mean follow-up in situ of 15.5 months and a maximum of 7.4 years. The probability of AlphaCor retention at 6 months and 1 and 2 years for protocol cases was 92%, 80%, and 62%, respectively, and off label cases were at higher risk (P = 0.010), as were cases not prescribed medroxyprogesterone (MPG; P = 0.001). Currently, the most common complications were stromal melting, fibrous reclosure of the posterior lamellar opening, and white intraoptic deposits, with incidences in 2005 of 11.4%, 5.1%, and 2.6%, respectively. MPG seems to protect against melts, and eyes with a history of herpetic keratitis were not at increased risk. A history of glaucoma or the presence of tubes did not affect device retention. Complications culminated in loss of an eye in 1.3%. Mean preoperative visual acuity (VA) was hand movements. The VA achieved postoperatively (light perception to 20/20) was affected by previous pathology and postoperative course, with a mean improvement of 2 lines. CONCLUSION: AlphaCor provides a treatment option where a donor tissue graft would not succeed in severe corneal conditions, while being reversible to a donor graft in the event of complications for anatomic integrity. Surgical technique and adjunctive therapies are evolving with experience. Continued data collection is important for a fuller understanding of AlphaCor's role. PMID- 17133050 TI - Randomized, double-masked clinical trial evaluating corneal endothelial cell loss after cataract extraction and intraocular lens implantation: Fluid-based system versus ultrasound phacoemulsification. AB - PURPOSE: To assess the endothelial cell loss after cataract extraction using a fluid-based system and compare it with conventional ultrasound. METHODS: A total of 46 eyes of 23 patients with up to 3+ bilateral cataract were included. The first eye was randomized to either fluid-based system or conventional ultrasound, and the fellow eye received the alternate system. Visual acuity and ultrasound pachymetry were measured preoperatively and at 1 day, 1 week, and 1 and 3 months postoperatively. Specular microscopy was performed preoperatively and 3 months after the surgery. Data was analyzed using the paired t test and Wilcoxon signed rank test. A P value less than 0.05 was considered statistically significant. RESULTS: Postoperatively, no significant differences were found among the groups. However, there was a significant difference (P < 0.01, analysis of variance with Tukey as posttest) in corneal pachymetry at day 1 after the surgery compared with baseline in both groups. There was a 6.3% and 7.9% loss of endothelial cells in the fluid-based system and ultrasound group, respectively. CONCLUSION: Visual acuity, pachymetry, and endothelial cell loss after cataract extraction and intraocular lens implantation using a fluid-based system is similar to conventional ultrasound. PMID- 17133051 TI - Effects of glaucoma medications on corneal endothelium, keratocytes, and subbasal nerves among participants in the ocular hypertension treatment study. AB - PURPOSE: To compare subbasal corneal nerve and keratocyte density and endothelial characteristics of ocular hypertensive patients treated with medications or observation. METHODS: Participants in the Ocular Hypertensive Treatment Study (OHTS) randomized at Mayo Clinic to medication or observation were evaluated with specular microscopy annually for 6 years. Confocal microscopy was performed 78 to 108 months after enrollment. Subbasal nerve density was calculated by manual tracing and digital image analysis. Keratocyte density was determined by manual counting methods. Data were compared using a t test and a rank sum test. RESULTS: After 6 years, corneal endothelial cell density, percent hexagonal cells, and coefficient of variation of cell area for the observation (n = 21) and medication groups (n = 26) were similar (2415 +/- 300 vs. 2331 +/- 239 cells/mm; 63% +/- 11% vs. 65% +/- 10%; and 0.32 +/- 0.07 vs. 0.30 +/- 0.06, respectively). Of 38 participants undergoing confocal examination, the medication group (n = 19) had fewer nerves (3.8 +/- 2.1 vs. 5.9 +/- 2.0 nerves/frame; P = 0.02) and a lower nerve density (5643 +/- 2861 vs. 9314 +/- 3743 mum/mm; P = 0.007) than the observation patients (n = 10). An additional 9 patients in the observation group, who began medication before confocal scanning, had intermediate nerve densities. Full-thickness keratocyte density was similar, with 22,257 +/- 2419 and 23,430 +/ 3285 cell/mm in the observation and medication groups, respectively. CONCLUSIONS: Chronic administration of glaucoma medications causes a decrease in the number and density of corneal subbasal nerve fiber bundles but does not affect keratocyte density or corneal endothelial characteristics. PMID- 17133052 TI - Efficacy of nonsimultaneous bilateral LASIK after nonsimultaneous bilateral penetrating keratoplasty. AB - PURPOSE: To show if nonsimultaneous bilateral laser in situ keratomileusis (LASIK) is a safe and effective procedure for patients with bilateral penetrating keratoplasty (PKP). METHODS: Five patients (10 eyes), with keratoconus, underwent PKP separately in each eye. After an average PKP follow-up of 45.8 months per eye, (range, 19-92 months), each eye underwent 1-stage LASIK using the Visx Star S3 laser to correct its residual refractive error. We used the cycloplegic refraction as the target for the LASIK surgery. Each patient had his or her eyes done separately 3 weeks apart. RESULTS: Pre-LASIK myopia averaged -3.93 +/- 2.9 D, ranging from -10.00 to +0.75 D. Pre-LASIK astigmatism averaged -3.25 +/- 0.80 D, ranging from -1.75 to -4.50 D. Mean pre-LASIK keratometry was 45.4 +/- 2.6, ranging from 42.2 to 50.5. Uncorrected visual acuity pre-LASIK averaged 20/220, ranging from 20/60 to 20/400. Best-corrected spectacle visual acuity (BCSVA) pre LASIK averaged 20/22, ranging from 20/20 to 20/30. Nine eyes had no complications. One eye had a flap buttonhole during LASIK, and surgery was aborted. No vision was lost in this eye. Mean follow-up after LASIK in the 9 eyes was 17 +/- 15.2 months, (range, 4-56 months). Mean post-LASIK ametropia in these 9 eyes was +0.25 +/- 0.45 D, ranging from -0.50 to +0.75 D. Average post-LASIK astigmatism was -0.33 +/- 0.38 D, ranging from 0 to -0.75 D. In the 9 treated eyes, uncorrected vision post-LASIK averaged 20/25, ranging from 20/20 to 20/30. BCSVA post-LASIK averaged 20/21, ranging from 20/20 to 20/25. No lines of visual acuity were lost in any of the eyes. CONCLUSION: Young patients who have had bilateral PKP, with good postoperative vision and low levels of myopia, astigmatism, and minimal wound override, are good candidates for bilateral nonsimultaneous LASIK. Further studies can now be done on the performance of bilateral simultaneous LASIK in patients who have had corneal transplant surgery in both of their eyes. PMID- 17133053 TI - Corneal cross-linking-induced stromal demarcation line. AB - PURPOSE: Corneal collagen cross-linking by UVA/riboflavin (X-linking) represents a new method for the treatment of progressive keratoconus and currently is under clinical study. To avoid UVA irradiation damage to the corneal endothelium, the parameters for X-linking are set in a way that effective treatment occurs only in the first 300 microm of the corneal stroma. Here, X-linking not only strengthens the biomechanical properties of the cornea but also induces keratocyte apoptosis. To date, the effectiveness of treatment could be monitored only indirectly by postoperative follow-up corneal topographies or using corneal confocal microscopy. Here we describe a corneal stromal demarcation line indicating the transition zone between cross-linked anterior corneal stroma and untreated posterior corneal stroma. The demarcation line is biomicroscopically detectable in slit-lamp examination as early as 2 weeks after treatment. METHODS: X-linking was performed in 16 cases of progressive keratoconus, and corneas were examined biomicroscopically and by means of corneal topography and pachymetry before and after treatment. RESULTS: In 14 of 16 cases, a thin stromal demarcation line was visible at a depth of approximately 300 microm over the whole cornea after X linking treatment. CONCLUSION: This newly observed demarcation line may result from differences in the refractive index and/or reflection properties of untreated versus X-linked corneal stroma and represents an effective tool to biomicroscopically easily monitor the depth of effective X-linking treatment in keratoconus. PMID- 17133054 TI - Modification in descemet-stripping automated endothelial keratoplasty: "Hitch suture" technique. AB - PURPOSE: To describe a new modification in Descemet-stripping automated endothelial keratoplasty, using a "hitch suture" to unfold the donor lenticule inside the recipient's eye. METHODS: The technique was used in 2 eyes of 2 patients with bullous keratopathy after complicated cataract surgery. A hitch suture was created in both the donor lenticules before inserting the folded graft in the recipient's anterior chamber. The hitch suture was pulled through a limbal stab incision to unfold the donor lenticule in both cases. RESULTS: No intraoperative problems were encountered. The grafts were well centered with corneal edema. The central part of the graft cleared over a period of 10 to 14 days. CONCLUSION: The technique of using a hitch suture to unfold the graft eliminates the problems associated with this difficult step of the surgery. The hitch loop can be easily made at the edge of the graft and can be pulled using a Sinskey hook through 1 of the stab incisions. We believe that this simple modification in this surgery would help simplify the learning curve of the procedure and increase its chances of success. PMID- 17133055 TI - Effect of hyperosmolality on beta-defensin gene expression by human corneal epithelial cells. AB - PURPOSE: As human beta-defensins (hBD) are important antimicrobial peptides at epithelial surfaces, including the ocular surface, we tested the effect of hyperosmolar conditions on the expression of these peptides by human corneal epithelial cells (HCECs). METHODS: Simian virus 40-transformed HCECs (n = 5) or primary cultured HCECs (n = 5) were treated with serum-free media or serum-free hyperosmolar (400-500 mOsm/kg) media for 24 hours or serum-free 500 mOsm/kg media for 12 to 48 hours. The effect of hyperosmolality on interleukin-1beta (IL-1beta) induced hBD-2 expression was also tested. IL-6 expression was studied as a marker of IL-1beta function. Expression of hBD-1, -2, and -3 and IL-6 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). The levels of active IL-1beta (culture supernatants and cell lysates) and pro-IL-1beta (cell lysates) were detected by enzyme-linked immunosorbent assay. RESULTS: HCECs constitutively expressed hBD-1 and -3 but not hBD-2. Hyperosmolar media had no effect on the basal expression of hBD-1 or -3 and did not induce the expression of hBD-2. Treatment with 500 mOsm/kg media for 24 hours decreased the ability of IL-1beta to upregulate hBD-2 and IL-6 expression. Active or pro-IL-1beta was not detected in any cell culture sample. CONCLUSION: Our results suggest that the hyperosmolar environment observed in diseases such as dry eye does not alter defensin expression. However, a hyperosmolar environment may influence cytokine function in ocular surface cells and thus affect their response to injury and inflammation. PMID- 17133056 TI - Immunoglobulin gene implicated in murine herpes stromal keratitis is not associated with the human disease. AB - PURPOSE: Susceptibility to herpes stromal keratitis (HSK) is strongly influenced by genetic factors, as shown by multiple rodent models using human herpes simplex virus. A single gene, encoding the immunoglobulin G (IgG) 2a heavy chain protein, confers susceptibility or resistance through a mechanism involving molecular mimicry in one mouse model. However, other rodent studies have produced contradictory results. This study tested the hypothesis that the GM23 gene (the human IgG2a homolog) influences susceptibility to HSK in humans. METHODS: The study population consisted of all consenting patients diagnosed with HSK (25 whites, 2 African Americans) at the Medical University of South Carolina Storm Eye Institute Clinic in Charleston, SC, between August 2000 and June 2004. Healthy controls (23 white adults with no history of HSK) were recruited from the same local population. Genomic DNA from subjects was genotyped at the GM23 locus, which has been implicated as an HSK resistance gene in animal models, by polymerase chain reaction-restriction fragment length polymorphism (RFLP) analysis. RESULTS: No difference in GM23 genotype frequency was observed between patients with HSK and controls. CONCLUSION: Susceptibility to HSK in whites is not predicted by GM23 genotype. PMID- 17133057 TI - Ethanol treatment induces significant cell death in porcine corneal fibroblasts. AB - PURPOSE: To explore the effect of ethanol treatment on corneal stromal cells. METHODS: Primary porcine corneal fibroblasts from passages 3 to 5 were treated with ethanol at concentrations of 10%, 15%, 20%, and 50% for 30 seconds. A control group was treated with phosphate-buffered saline (PBS) for 30 seconds. Morphologic changes were documented with phase-contrast microscopy, and the growth curves were examined with a PicoGreen assay. Cellular viability was examined with an ethidium homodimer and calcein-AM stain, whereas cellular apoptosis and/or necrosis were analyzed by a YO-PRO-1 dye/propidium iodide apoptosis assay coupled with flow cytometry and further confirmed with a genomic DNA pattern assay. Cellular toxicity was examined with a lactate dehydrogenase (LDH) assay. RESULTS: Significant cell rounding and detachment from the culture dish were noticed after 20% ethanol treatment of 30 seconds, despite that the cell morphology remained unchanged in the PBS and 10% and 15% ethanol groups. Twenty percent ethanol induced significant cellular toxicity, causing cell death as shown by ethidium homodimer and calcein-AM stain, YO-PRO-1 dye/propidium iodide apoptosis assay, and LDH assay, although 10% and 15% ethanol caused minimal changes to corneal fibroblasts. Cellular death was most significant 6 hours after the 20% ethanol treatment. The genomic DNA pattern revealed intact DNA in the control, 10% ethanol, and 15% ethanol groups at all times, whereas DNA smearing was noticed at 48 hours after the 20% ethanol treatment. However, none of the DNA examined revealed significant DNA laddering patterns of apoptosis. Fifty percent ethanol treatment of 30 seconds resulted in cell fixation and cell death. CONCLUSION: Treatment with 20% ethanol for 30 seconds induced significant porcine corneal fibroblast cell death, whereas 10% and 15% ethanol treatment of 30 seconds caused minimal changes. We propose that, when applied for 30 seconds, 20% ethanol is the threshold level that causes cell death in cultured porcine corneal fibroblasts. PMID- 17133058 TI - Measurement of light transmittance of cryopreserved human amniotic membrane. AB - PURPOSE: To measure the light transmission of human amniotic membrane (AM) and compare it with that of therapeutic soft contact lens (TSCL). METHODS: Total light transmittance of each sample (4 AMs and 2 TSCLs) was measured in the 250- to 800-nm range by using a spectrophotometer. RESULTS: The percent light transmission of TSCL was nearly 100% at wavelengths greater than 450 nm, decreased gradually in the sub-450-nm range, and reached 78.4 +/- 3.7 at 250 nm. In contrast, the percent transmission of the 4 AMs decreased gradually and continually throughout the spectrum as wavelength shortened. The transmission of light through AM dropped to less than 20% at 250 nm but remained 90% or more at wavelengths more than 600 nm. CONCLUSION: Light transmittance properties of cryopreserved human AM and human cornea are relatively similar, but AM (and not cornea) allows transmission of 24.6% of UVC. PMID- 17133059 TI - Efficacy and safety of moxifloxacin as an additive in Optisol-GS a preservation medium for corneal donor tissue. AB - PURPOSE: To assess the endothelial toxicity and the microbiological efficacy of moxifloxacin (250 microg/mL) as an additive to Optisol-GS. METHODS: Five hundred nine donor rims were studied. One half of each donor rim was placed in standard Optisol-GS and the other half of the rim in Optisol-GS fortified with moxifloxacin (250 microg/mL). All rims were refrigerated for 24 hours at 3 degrees C and placed in thioglycolate broth and incubated at 37 degrees C for 7 days. One pair of donor buttons not used in transplantation stored in each solution was examined for endothelial changes by using electron microscopy (EM). A second pair of cornea buttons was examined for toxicity by endothelial staining with 0.3% trypan blue and 0.2% alizarin red. All endothelial cells that stained (nonviable cells) and nonstained cells (viable cells) were counted, and the ratio of nonviable cells was calculated. RESULTS: The rate of culture-positive donor rims in the Optisol-GS group was 11.9% (61/509) and in the moxifloxacin-fortified Optisol-GS media was 2.5% (13/509). The difference was statistically significant (P < 0.01; chi test). There was no difference in the cellular morphology of the button stored in moxifloxacin-fortified Optisol-GS compared with Optisol-GS using EM. In the bioassay, the rate of nonviable cells in the moxifloxacin-fortified media compared with the control media was nonsignificant (P > 0.05). CONCLUSION: Moxifloxacin (250 microg/mL) seems to be safe as an additive agent for cornea storage media. It significantly reduces the rate of positive rim cultures and shows no signs of endothelial cytotoxicity as viewed by EM and by a bioassay of trypan blue and alizarin red. PMID- 17133060 TI - Corneal trephination with the femtosecond laser. AB - PURPOSE: To evaluate the feasibility and cut quality of corneal trephination in human donor corneal tissue with the femtosecond laser. METHODS: Twelve human corneoscleral discs were inserted in an artificial anterior chamber. After corneal thickness measurement and tonometry, the cornea was mounted on a femtosecond laser (FEMTEC; 20/10 Perfect Vision, Heidelberg, Germany) through a contact lens (patient interface). Trephination was performed with diameters of 7.0, 7.5, 8.0, and 8.5 mm in 3 corneas each. The corneal button was removed from the corneoscleral disc in 2 of the 3 corneas in each case. The cut was not manipulated in the remaining corneas to enable histologic detection of possible tissue bridges. The cut edges were macroscopically and light-microscopically examined for quality. RESULTS: Corneal buttons and corneoscleral discs could be separated by blunt dissection in all cases. Tissue bridges were more common in thicker edematous corneas than in thinner ones. Both the macro- and microscopic examination disclosed smooth rectilinear cut margins with a perpendicular cut edge. CONCLUSION: This feasibility study shows that the femtosecond laser enables sufficient trephination of human donor corneas. PMID- 17133061 TI - Histology of posterior lamellar keratoplasty. AB - PURPOSE: To report histology of a posterior lamellar graft performed to treat clinically significant endothelial disease in a patient with Fuchs dystrophy. METHODS: A 78-year-old patient with Fuchs dystrophy underwent posterior lamellar keratoplasty (PLK), the original form of endothelial keratoplasty performed with manual lamellar dissection of the recipient and donor corneas. Eighteen months later, the patient had herpes simplex virus keratitis and graft rejection episodes. One year after the infection resolved (2.5 years after PLK), penetrating keratoplasty was performed, and the excised corneal button was examined. RESULTS: Histopathologic evaluation of the corneal button showed no discernible interface, opacity, or gapping between the anterior host stromal tissue and PLK donor tissue or any signs of significant migration of donor endothelial cells onto recipient tissue. CONCLUSION: This PLK histology showed firm attachment with no visible interface between the posterior donor and anterior host stromal tissue. Surprisingly, no significant donor endothelial cell migration was detected on adjacent recipient tissue where Descemet membrane had been excised or where endothelial cells were lost as a result of the procedure. PMID- 17133062 TI - Superficial hypertrophic dendritic epitheliopathy occurring before and after penetrating keratoplasty in a patient with psoriasis. AB - PURPOSE: To report a patient with superficial hypertrophic dendritic epitheliopathy that occurred in 1 eye after keratoplasty and in the other eye that had not undergone previous corneal transplantation. In both eyes, the epitheliopathy recurred even after subsequent penetrating keratoplasties. METHODS: Case report. RESULTS: A patient with chronic eyelid and ocular surface disease secondary to psoriasis was monitored over a 17-year period. During this time, the patient developed a superficial hypertrophic dendritic epitheliopathy of the left eye 3 years after corneal transplantation. In the right eye, which had not undergone a penetrating keratoplasty, the patient was also noted to have a superficial hypertrophic dendritic epitheliopathy that changed morphologies over the ensuing 5.5 years. Both eyes subsequently required penetrating keratoplasties for decreased vision secondary to subepithelial scarring. After surgery, both corneas suffered recurrences of the superficial hypertrophic epitheliopathy. CONCLUSION: In addition to the original findings that superficial hypertrophic dendritic epitheliopathy occurs in postkeratoplasty eyes, this condition can also occur in corneas that have not been transplanted previously. The morphology of these lesions can vary. PMID- 17133063 TI - Corneal graft rejection precipitated by uveitis secondary to alendronate sodium therapy. AB - PURPOSE: To report a case of corneal graft rejection precipitated by severe uveitis secondary to alendronate therapy and to review the literature of relevance to this case. METHODS: A 77-year-old woman with a hypopyon and corneal graft rejection was studied for possible precipitants, including herpes viral and bacterial infection. Results were negative. She was treated unsuccessfully with systemic and topical steroids, systemic antivirals, and intraocular antibiotic therapy. RESULTS: Withdrawal of alendronate resulted in rapid resolution of intraocular inflammation and corneal edema. CONCLUSION: We recommend vigilance in corneal transplant patients on simultaneous bisphosphonate therapy. Caution is advised in the extension to human trials of animal studies investigating the use of bisphosphonates in corneal transplantation. PMID- 17133064 TI - Confocal microscopy of subepithelial infiltrates occurring after epidemic keratoconjunctivitis. AB - PURPOSE: To describe the confocal microscopic findings in subepithelial infiltrates associated with epidemic keratoconjunctivitis (EKC). METHODS: Observational case report. RESULTS: A 14-year-old Saudi girl with keratoconus developed subepithelial infiltrates (SEIs) after the onset of bilateral EKC. Confocal microscopy of the left cornea 8 weeks after the onset of EKC showed many highly reflective dendritic cells at the level of basal epithelium, epithelial basement membrane zone, and anterior stroma, as well as many highly reflective fusiform and round cells within the anterior stroma, with deceasing density in progressively deeper layers of the stroma. These findings were not present on confocal microscopy that had been performed 2 weeks before the onset of EKC. CONCLUSION: Confocal microscopic examination of SEIs after EKC provides evidence of an inflammatory response localized to the basal epithelium and anterior stroma of the central cornea. PMID- 17133065 TI - Filamentary keratopathy caused by corneal occlusion by large-angle paralytic strabismus. AB - PURPOSE: To report a case of filamentary keratopathy in a patient with large angle paralytic strabismus. METHODS: A 31-year-old man who had a traffic accident was diagnosed to have third cranial nerve palsy and a large-angle exotropia in his right eye. The patient complained of foreign body sensation in his right eye 2 months after the accident. RESULTS: Slit-lamp examination showed multiple filaments on the right cornea. Debridement of the filaments and artificial tears relieved the symptoms and were used until the paralysis resolved. CONCLUSION: Large-angle paralytic strabismus with ptosis and poor elevation and depression of the eye may occlude the cornea and lead to filamentary keratopathy. PMID- 17133066 TI - Late-onset traumatic dislocation with central tissue loss of laser in situ keratomileusis flap. AB - PURPOSE: We report the occurrence and outcome of a severe late-onset traumatic dislocation of a laser in situ keratomileusis (LASIK) flap with loss of central flap tissue. METHODS: Case report. RESULTS: A 40-year-old woman underwent uncomplicated bilateral LASIK surgery, followed 5 years later by an enhancement procedure in both eyes. Ocular trauma with a power sander occurred 6 years after LASIK and 1 year after the enhancement procedure. The flap was found to have an almost complete tear from the nasal hinge and a central tissue defect. After irrigating, repositioning, and stabilizing the flap with 2 nylon sutures, a bandage contact lens was placed. The patient was treated with topical antibiotic and steroid drops. Stage 2 diffuse lamellar keratitis developed, which responded to topical treatment. Ten weeks after injury, the patient regained an uncorrected visual acuity of 20/20-1. CONCLUSION: Late-onset trauma to the LASIK flap can result in flap dehiscence and tissue loss. Prompt and appropriate management can lead to an excellent visual outcome even in severely traumatized dislocated LASIK flaps. PMID- 17133067 TI - Corneal myxoma arising in a patient with repeated phototherapeutic keratectomies. AB - PURPOSE: We report the clinical and pathologic findings of a corneal myxoma that was diagnosed in a cornea that had undergone phototherapeutic keratectomy (PTK). METHODS: A corneal myxoma was identified in a full-thickness keratoplasty specimen obtained from a 51-year-old man with a history of a forceps injury to his right eye at birth. The patient had presented with corneal edema and bullous keratopathy, which was treated on 3 occasions with PTK, resulting in progressive, persistent milky-white clouding of the cornea. This represents the eighth case of corneal myxoma in the reviewed literature and the first described after PTK. Thorough histopathologic and ultrastructural analyses were conducted. RESULTS: The histochemical, immunohistochemical, and electron microscopic findings were consistent with previously reported corneal myxomas. Most of the reported myxomas arising in the cornea presented with a history of prior corneal disease or injury and showed absence or disruption of Bowman layer histopathologically, as was found in our patient. CONCLUSION: The pathogenesis of corneal myxomas involves a reactive process that requires an antecedent affliction, in contrast to the neoplastic mesenchymal histogenesis of myxomas characterized in other parts of the body. PMID- 17133068 TI - Treatment of epithelial ingrowth after LASIK enhancement with a combined technique of mechanical debridement, flap suturing, and fibrin glue application. AB - PURPOSE: To report a case of clinically significant post-laser in situ keratomileusis (LASIK) epithelial ingrowth successfully treated with a combined technique of mechanical debridement, flap suturing, and fibrin glue application. METHODS: A retrospective case report. RESULTS: A 42-year-old female patient underwent LASIK and an enhancement procedure in 1998 and 2002, respectively. Two years after her enhancement, she developed severe, visually significant epithelial ingrowth. Treatment was undertaken using a combination of mechanical debridement, flap suturing, and fibrin glue application. No recurrence was found during a 15-month follow-up period. No adverse effects were seen with this approach. CONCLUSION: Severe progressive epithelial ingrowth may be treated successfully with a combination of mechanical debridement, flap suturing, and fibrin glue application. PMID- 17133069 TI - Mycobacterium tuberculosis presenting as chronic red eye. AB - PURPOSE: To report a case of tuberculosis of the conjunctiva. METHODS: Case report with pathologic correlation. A 17-year-old man who had relocated to Australia from Liberia presented with chronic unilateral conjunctivitis. RESULTS: The diagnosis was not initially evident, despite 2 separate biopsy specimens, conjunctival cultures, and polymerase chain reaction testing. Definitive diagnosis was made after repeated histologic examination. Antituberculous treatment resulted in prompt resolution of all ocular signs. CONCLUSION: Tuberculous conjunctivitis is now a very rare condition in the developed world. Definitive diagnosis requires the identification of Mycobacterium tuberculosis organisms in conjunctival biopsy specimens-either through microscopic detection of acid-fast bacilli or more sensitive culture techniques. PMID- 17133070 TI - Conjunctival malignant melanoma-associated pseudomelanoma of the apposing tarsal conjunctiva. AB - PURPOSE: To describe a case of conjunctival malignant melanoma associated with pseudomelanomatous alteration of the apposing tarsal conjunctiva. METHODS: A 93 year-old woman presented with an elevated, pigmented mass on her superior bulbar conjunctiva. The tumor was associated with increased pigmentation of the apposing superior tarsal conjunctiva. An excisional biopsy of the epibulbar melanoma and pigmented tarsal conjunctiva was performed. RESULTS: Histopathologic evaluation of the epibulbar tumor revealed epithelioid melanocytes diagnostic of malignant melanoma. Histopathologic evaluation of the pigmented tarsal lesion showed large areas of ulceration and foci of granulation tissue composed of neovascular sprouts arising in the background of a loose connective tissue, with a moderate chronic inflammatory infiltrate. The infiltrate was composed of mature lymphocytes, plasma cells, and scattered histiocytes. Densely packed intracytoplasmic, brown pigment granules that stained positive with HMB 45 were found, attesting to adjacent melanocytes' releasing melanin-laden granules. There was no evidence of malignancy in the pigmented tarsal specimen. CONCLUSION: Noncontiguous pigmented conjunctival tumor can be found in apposition to an epibulbar melanoma. Although a limited biopsy could be considered, only a complete resection and histopathologic evaluation can determine whether the entire lesion is free of malignant melanoma. PMID- 17133071 TI - Diagnosis of epithelial ingrowth after penetrating keratoplasty with confocal microscopy. AB - PURPOSE: To report confocal microscopy use in the clinical diagnosis of epithelial ingrowth after penetrating keratoplasty (PKP). METHODS: A 36-year-old female patient with keratoconus developed a well-delimited posterior hazy membrane covering the inferior two thirds of the cornea 3 months after an uneventful PKP. A posterior corneal line was present resembling an endothelial graft rejection line, but with no keratic precipitates or corneal edema. Ocular hypertension was not observed. Confocal microscopy was performed to elucidate the diagnosis. RESULTS: Confocal microscopy showed epithelium and stroma with normal findings. Two distinct cellular types were presented at the endothelium layer. Enlarged endothelial cells were observed in the superior part of the cornea up to the leading edge of the hazy membrane. In the middle and inferior part of the graft, the cells were larger, with polygonal shape and easily recognizable hyperreflective nuclei, suggestive of epithelial cells. With these confocal microscopy findings, the patient was promptly submitted to another PKP. Histologic analysis confirmed the diagnosis of epithelial ingrowth. CONCLUSION: Confocal microscopy imaging technique seems to be a useful tool in the early diagnosis of epithelial ingrowth after PKP. PMID- 17133072 TI - Rothia dentocariosa isolated from a corneal ulcer. AB - PURPOSE: Rothia dentocariosa is a common commensal in the oropharyngeal cavity but a rare human pathogen. Ocular culture has been documented only twice previously, both from vitreous samples taken in the context of endophthalmitis. These cases, and other reports of human Rothia infection, have proposed hematogenous spread from the oropharynx as the mode of transmission. METHODS: A case report of an 11-year-old boy with a progressive right corneal abscess that required penetrating keratoplasty because of corneal perforation is detailed. The keratitis recurred in the graft, leading to an almost total epithelial defect, hypopyon, and descemetocele within 3 months. RESULTS: R. dentocariosa was eventually isolated from the cornea, and the patient made a rapid recovery once topical medication was altered accordingly. Microbial identification was confirmed at a reference laboratory by using partial sequencing of 16s rDNA. The father later described his son's habit of wetting the fingertip with saliva before eyelid rubbing as a means of reducing ocular discomfort. CONCLUSION: This is the first reported case of corneal isolation of R. dentocariosa. It also suggests a direct mode of transmission of the organism to the eye by contaminated saliva. PMID- 17133074 TI - Effect of the fourth-generation fluoroquinolones on corneal reepithelialization after penetrating keratoplasty. PMID- 17133076 TI - Gut microbes, the innate immune system and inflammatory bowel disease: location, location, location. PMID- 17133077 TI - Update on Clostridium difficile associated disease. AB - PURPOSE OF REVIEW: The aim of this article is to report recent changes in the epidemiology of Clostridium difficile associated disease. RECENT FINDINGS: An epidemic of Clostridium difficile associated disease in Quebec was associated with a threefold increase in incidence and a sharp increase in fatalities. Strain typing of C. difficile isolates from the involved hospitals revealed that 82% were of a single strain (NAP1/027). This strain was found to produce greater than 10 times as much toxin A and toxin B as historic isolates and has been identified in many institutions throughout North America and Europe. Frequent nosocomial use of fluoroquinolones may encourage the spread of this strain as it is fluoroquinolone resistant. An increased rate of community-acquired Clostridium difficile-associated disease has also been noted and, in some cases, without prior antibiotic exposure. Although some studies have suggested an increased failure rate of metronidazole in Clostridium difficile associated disease, it remains the recommended first line treatment for uncomplicated cases. Other antibiotics, a toxin binder, probiotic agents and a vaccine are being tested in clinical trials for efficacy in prevention and treatment of Clostridium difficile associated disease. SUMMARY: The recent increase in the incidence and severity of Clostridium difficile associated disease may be related, at least in part, to the emergence of a highly virulent, fluoroquinolone-resistant, NAP1/027 strain. PMID- 17133078 TI - Quorum sensing by enteric pathogens. AB - PURPOSE OF REVIEW: This review presents advances in our understanding of how pathogenic, enteric bacteria use quorum sensing to regulate several traits that allow them to establish and maintain infection in their host, including motility, biofilm formation, and virulence-specific genes. RECENT FINDINGS: Quorum sensing in enteric bacteria has been elusive for a long time. Recent data indicate that enteric bacteria use several quorum-sensing mechanisms including the LuxR-I quorum-sensing system, the LuxS/AI-2 system, and the AI 3/epinephrine/norepinephrine system to assess their environment and to recognize the host environment. These systems allow bacteria to communicate across species boundaries, and the AI-3/epinephrine/norepinephrine system is involved in interkingdom signaling. SUMMARY: Recent developments in our understanding of the molecular and biochemical mechanisms involved in quorum sensing as well as the chemical signal(s) to which bacteria respond and their corresponding physiological responses will improve our understanding of bacterial pathogenesis and microbial flora-host interactions, and potentially lead to novel strategies for combating infection. PMID- 17133079 TI - Adherent-invasive Escherichia coli and Crohn's disease. AB - PURPOSE OF REVIEW: The identification of mutations in the nucleotide oligomerization domain 2-encoding gene in patients with Crohn's disease suggests a link between the innate immune response to invasive bacteria and the development of Crohn's disease. Herein, we review reports concerning the association of pathogenic Escherichia coli with the intestinal mucosa of Crohn's disease patients. RECENT FINDINGS: Adherent-invasive E. coli were isolated from ileal biopsies of 36.4% of patients with ileal Crohn's disease. Adherent-invasive E. coli colonize the intestinal mucosa by adhering to intestinal epithelial cells. They are also true invasive pathogens, able to invade intestinal epithelial cells and to replicate intracellularly. Adherent-invasive E. coli strains replicate extensively into macrophages inducing the secretion of very large amounts of tumor necrosis factor-alpha. Similar pathogenic E. coli strains were recently associated with granulomatous colitis of Boxer dogs. Interestingly, high levels of E. coli outer membrane protein C antibodies are present in 37-55% of Crohn's disease patients and reactivity to outer membrane protein C is associated with increased severity of Crohn's disease. SUMMARY: As the infection cycle of adherent-invasive E. coli could depend upon the ability of these pathogenic bacteria to colonize the gastrointestinal tract of genetically predisposed Crohn's disease patients, antibiotics which could eradicate the bacteria, or probiotics which could substitute them in the gastrointestinal tract, could be of therapeutic value in ileal Crohn's disease. PMID- 17133080 TI - The role of altered microbial signaling via mutant NODs in intestinal inflammation. AB - PURPOSE OF REVIEW: Recently, a new class of intracellular pattern recognition receptors belonging to the family of nucleotide binding and oligomerization domain (NOD)-like receptors that includes NOD1, NOD2 and IPAF has been described. These proteins are involved in recognizing bacterial components or their degradation constituents that are delivered within the cytoplasm. In this review we will analyze the role of NOD proteins in regulating immune homeostasis. RECENT FINDINGS: After an initial description of advances in our understanding of the function of these proteins, this review will focus on the contradictory finding that even though mutations in NOD2 proteins lead to a loss of function phenotype, the outcome is an increased inflammatory response. Different hypotheses to reconcile this observation will be proposed. SUMMARY: The cellular and tissue distribution of NOD molecules as well as their role in regulating inflammatory cytokine release renders these proteins particularly important in controlling the development of inflammatory reactions. This is confirmed by the discovery that mutations in the genes that code for NOD1 and NOD2 confer increased susceptibility to inflammatory bowel disease. We will discuss NOD2 involvement in the development of Crohn's disease. PMID- 17133081 TI - Toll-like receptor signaling in intestinal epithelial cells contributes to colonic homoeostasis. AB - PURPOSE OF REVIEW: Since intestinal epithelium expresses Toll-like receptors, it was suggested that the intestinal epithelium is actively involved in the maintenance of colonic homeostasis. Here we describe our recent findings, which support an active contribution of colonic epithelium to intestinal homeostasis via a unique activation of epithelial TLR9. RECENT FINDINGS: Recent data indicate that stimulation of Toll-like receptors by intestinal microbiota supports colonic homeostasis. Several Toll-like receptors are expressed in intestinal epithelium. TLR9, an intracellular protein in immune cells, is expressed on the cell surfaces of intestinal epithelium, both on the apical and the basolateral membrane. TLR9 signaling varies in a domain-specific manner; whereas JNK is activated by TLR9 ligand both apically or basolaterally, NF-kappaB is activated only via basolateral stimulation. In apical TLR9 stimulation, IkappaB is phosphorylated and ubiquitinated but is not degraded, and NF-kappaB-dependent inflammatory signals are not transduced. Stimulation of apical TLR9 compromises the inflammatory cascade induced basolaterally by several other Toll-like receptor ligands, suggesting that apical exposure to luminal microbial DNA restrains intestinal inflammation. SUMMARY: These data indicate that certain luminal bacterial products support colonic homeostasis via activation of epithelial Toll like receptors. The role of epithelial Toll-like receptor expression and activation in the pathogenesis of human inflammatory bowel disease is yet to be explored. PMID- 17133082 TI - Defensins and cathelicidins in gastrointestinal infections. AB - PURPOSE OF REVIEW: To review recently published studies presenting novel and relevant information on antimicrobial peptides in gastrointestinal infections. RECENT FINDINGS: Defensins and cathelicidins are important antimicrobial peptides expressed by the gastrointestinal epithelium. Their localization and regulation have been the focus of current research establishing the relevance of these peptides both in counteracting an attack by pathogens as well as in controlling the endogenous bacterial flora. In the small intestine, Paneth cell alpha defensins maintain a low level of microorganisms and regulate the composition of the bacterial flora. In contrast, a constitutive beta-defensin can be found in nearly all gastrointestinal tissues. Other relevant beta-defensins as well as human cathelicidin are inducible by inflammation or infections. Thus Helicobacter pylori enhances defensin expression in the gastric mucosa and Campylobacter jejuni and Salmonella provoke a similar response in the colon. Other pathogenic bacteria may suppress the antimicrobial peptide response as an escape strategy. Notably, the therapeutic induction of cathelicidins alleviates experimental shigellosis, suggesting a future role of endogenous antibiotics in medical therapy. SUMMARY: These recent findings together with a better understanding of underlying mechanisms involved in the regulation and biology of antimicrobial peptides will open up new therapeutic avenues to battle infections. PMID- 17133083 TI - Rotavirus: to the gut and beyond! AB - PURPOSE OF REVIEW: Rotavirus causes severe gastroenteritis in children. A principle of rotavirus pathogenesis has been that the infection remains localized to epithelial cells in the small intestine. This dogma was challenged by recent findings of rotavirus in the serum of experimentally infected animals and children with diarrhea. Repeated associations of rotavirus infections with a wide range of nongastroenteric clinical manifestations in humans were considered spurious because of lack of proof that rotavirus escaped the intestine. New data outlined in this review, however, show that rotavirus routinely infects systemically and highlight controversies and future research questions. RECENT FINDINGS: Rotavirus antigens (antigenemia), RNA, or infectious virus (viremia) has been demonstrated in the serum and many extraintestinal tissues in all experimental animal models. Rotavirus antigens and RNA have been detected in the sera of children with rotavirus diarrhea. The tissues and cell types that support rotavirus replication outside the intestine and the consequences of extraintestinal reservoirs of infection are beginning to be examined. SUMMARY: Rotavirus infection is systemic, with an acute active viremia and extraintestinal replication. The impact of systemic rotavirus on disease burden remains to be determined. PMID- 17133084 TI - Chemoprevention of colorectal neoplasia: advances and controversies (the COX-2 story). AB - PURPOSE OF REVIEW: Colorectal cancer is prevalent, deadly and expensive to treat. Chemoprevention provides an opportunity to prevent colorectal cancer through prevention of precursor adenomas. RECENT FINDINGS: Three prospective randomized trials demonstrate significant reductions in new sporadic adenoma formation in individuals taking COX-2 selective inhibitors compared to placebo, but with an associated increase in cardiovascular risk. SUMMARY: Chemoprevention of colorectal adenomas with COX-2 selective nonsteroidal anti-inflammatory drugs is feasible, but these agents are unlikely to be used routinely for prevention of sporadic adenomas due to associated cardiovascular risk. Their use in other groups will depend on the answers to several questions regarding mechanisms of action and risk versus benefit. PMID- 17133085 TI - Colon cancer, fatty acids and anti-inflammatory compounds. AB - PURPOSE OF REVIEW: To outline recent findings on the efficacy of n-3 polyunsaturated fatty acids in the prevention/treatment of inflammatory bowel disease and colorectal cancer. RECENT FINDINGS: Compelling data indicate a functional link between chronic inflammation and colon cancer. With respect to environmental risk factors, there is growing evidence that long-chain n-3 polyunsaturated fatty acids found in fish oil suppress inflammatory bowel diseases and colon cancer risk in humans. Unfortunately, the molecular basis of the effect of n-3 polyunsaturated fatty acids on inflammation/colitis-associated colon cancer risk is still largely obscure. In this review, we focus on recent studies which address three emerging mechanisms of n-3 polyunsaturated fatty acids action: (1) metabolic interconversion into bioactive eicosanoids, (2) modulation of nuclear receptor activation, and (3) alteration of membrane phospholipid composition and functionality of lipid microdomains. SUMMARY: The consumption of dietary fish oil may prove to be an effective adjuvant therapy in colon cancer. Therefore, it is both appropriate and timely to determine precisely how n-3 polyunsaturated fatty acids modulate cell signaling networks, and reduce the risk of developing colon cancer and inflammatory disorders of the intestine. PMID- 17133086 TI - The use of nitric oxide-donating nonsteroidal anti-inflammatory drugs in the chemoprevention of colorectal neoplasia. AB - PURPOSE OF REVIEW: Nitric oxide-donating nonsteroidal anti-inflammatory drugs are emerging as a promising class of compounds for the chemoprevention of colon cancer. Recent progress in their preclinical and mechanistic evaluation is reviewed. RECENT FINDINGS: Compared to their parent compounds, nitric oxide donating nonsteroidal anti-inflammatory drugs are up to several hundred times more potent in inhibiting the growth of colon cancer cell lines and also quite effective in preventing colon cancer in various tumor animal models. Their chemopreventive effect is brought about through a strong cell kinetic effect, including inhibition of proliferation, induction of cell death and inhibition of cell cycle phase transitions. The induction of oxidative stress appears mechanistically crucial. Pleiotropic effects on cell signaling have been identified including Wnt, NOS2, mitogen-activated protein kinase and Nrf2 signaling. Nitric oxide-donating nonsteroidal anti-inflammatory drugs, particularly nitric oxide-donating aspirin, appear to be very safe compounds, as evidenced from many animal and early human studies. SUMMARY: Nitric oxide donating nonsteroidal anti-inflammatory drugs hold the promise of being safe and effective chemopreventive agents against colon cancer. Clinical trials are needed to determine whether these drugs can be applied clinically. PMID- 17133087 TI - Pharmacogenetics and diseases of the colon. AB - PURPOSE OF REVIEW: The deciphering of the human genome sequence has enabled the identification of genetic polymorphisms that are responsible for inter-individual variation in the response to drug therapy. This field is referred to as pharmacogenetics. We review the impact of pharmacogenetics on therapy in diseases of the colon using three common variant enzyme systems as examples. RECENT FINDINGS: Many enzyme systems impact the treatment of diseases of the colon. Examples include thiopurine S-methyltransferase, dihydropyrimidine dehydrogenase and flavin monooxygenase 3. They affect the management of inflammatory bowel disease, colorectal cancer and the chemoprevention of colorectal adenoma by influencing the metabolism of their respective substrates, azathioprine/6 mercaptopurine, 5-fluorouracil and sulindac. Recent studies have implicated the significance of genetic polymorphisms in each of the three drug-metabolizing enzymes, which impacts on the therapeutic outcome of the stated diseases. These studies highlight the potential role of pharmacogenetics in the design of a therapeutic plan which would increase efficacy and limit toxicity. SUMMARY: Pharmacogenetics of drug-metabolizing systems continues to gain significance in the drug therapy of a variety of disease states including those of the gastrointestinal tract. PMID- 17133088 TI - Recent developments in the role of endoscopic ultrasonography in diseases of the colon and rectum. AB - PURPOSE OF REVIEW: Endoscopic ultrasound has evolved as a useful technique for imaging and intervention in a variety of gastrointestinal and extraintestinal diseases including diseases of the colon and rectum. This paper will review recent developments in endoscopic ultrasound for colorectal diseases. RECENT FINDINGS: Recent studies have shown significant clinical impact of endoscopic ultrasound in rectal cancer staging. Iliac lymph node evaluation by endoscopic ultrasound-guided fine needle aspiration may further expand the role of endoscopic ultrasound in rectal cancer. Three-dimensional endoscopic ultrasound may help decrease some of the errors of staging with two-dimensional endoscopic ultrasound and may further improve staging accuracy. Recent studies have confirmed continued problems with re-staging rectal cancer after chemoradiation. Endoscopic ultrasound-fine needle aspiration can be helpful in detecting local recurrence of rectal cancer and has been shown to be useful in evaluation of subepithelial masses of the colon and rectum and evaluation of rectosigmoid endometriosis. SUMMARY: Endoscopic ultrasound continues to be useful for a variety of conditions of the colon and rectum with recent studies confirming its clinical impact as well as expanding its role into newer indications. Assessment for residual cancer after chemoradiation is still problematic and hopefully technological developments in ultrasound in the future may help in improving the accuracy of endoscopic ultrasound in this situation. PMID- 17133089 TI - Large bowel carcinoid tumors. AB - PURPOSE OF REVIEW: Gastrointestinal carcinoids comprise 90% of all carcinoid tumors and all carcinoids have malignant potential. This review focuses on the morphology, prognosis, detection and treatment for appendiceal, colonic and rectal carcinoids. RECENT FINDINGS: Computed tomography exists as an initial examination for the primary carcinoid tumor as well as for metastases. There have, however, been recent developments of newer and more accurate modalities. Octreotide scanning has a sensitivity of primary tumor detection of 90%; additionally, I- or I-meta-iodobenzylguanidine scanning and tracer-specific positron emission tomographic scans have demonstrated encouraging results. Further advancements in treatment with tumor-targeted therapy and biochemical evaluation of carcinoids have shown promise. SUMMARY: The recent progress with scintigraphic and radiologic modalities has provided better means for diagnosis of primary and metastatic carcinoid tumors. These newly discovered diagnostic modalities have been more encouraging than the recent treatment approaches that have been studied with regard to metastatic carcinoids. While surgery remains the mainstay of treatment of nonmetastatic carcinoid, there have been studies for various medical treatments of metastatic disease. Unfortunately, there have been disappointing results with regard to improvement of tumor response and patient survival, but a foundation has been established for future trials employing alternative agents and exploration of combination therapies. PMID- 17133090 TI - Bibliography. Current world literature. Gastrointestinal infections. PMID- 17133093 TI - Regulation of neutrophil homeostasis. AB - PURPOSE OF REVIEW: Neutrophils are an essential component of the innate immune response and a major contributor to inflammation. Consequently, neutrophil number in the blood is tightly regulated. Herein, we review recent studies that have greatly advanced our understanding of the mechanisms controlling neutrophil homeostasis. RECENT FINDINGS: Accumulating evidence shows that stromal derived factor-1 (CXCL12) through interaction with its major receptor CXCR4 provides a key retention signal for neutrophils in the bone marrow. Granulocyte colony stimulating factor induces neutrophil release from the bone marrow, in major part, by disrupting stromal derived factor-1/CXCR4 signaling. Granulocyte colony stimulating factor expression is regulated by a novel feedback loop that senses neutrophil emigration into tissues. Specifically, engulfment of apoptotic neutrophils by tissue phagocytes initiates a cytokine cascade that includes interleukin-23, interleukin-17, and ultimately granulocyte colony-stimulating factor. SUMMARY: Granulocyte colony-stimulating factor plays a central role in the dynamic regulation of neutrophil production and release from the bone marrow in response to environmental stresses. Recent studies have begun to elucidate both the pathways linking neutrophil clearance to granulocyte colony-stimulating factor expression and the mechanisms by which the factor induces neutrophil release from the bone marrow. These studies may lead to novel strategies to modulate neutrophil responses in host defense and inflammation. PMID- 17133094 TI - The negative regulatory roles of suppressor of cytokine signaling proteins in myeloid signaling pathways. AB - PURPOSE OF REVIEW: Suppressor of cytokine signaling proteins are key regulators of the response of myeloid cells to cytokines and other extracellular stimuli. This review explores recent developments that have shed light on how two of the best-characterized suppressor of cytokine signaling proteins, SOCS1 and SOCS3, attenuate myeloid signaling pathways that lead to inflammation, hematological malignancy, and related disorders. RECENT FINDINGS: In addition to its 'classic' role as an inhibitor of IFNgamma signaling, a number of new regulatory roles in IFNalpha and toll-like receptor signaling have been defined for SOCS1, and substantial progress has been made in identifying the factors that give rise to lethal inflammation in Socs1 neonatal mice. The aberrant transcriptional regulation of suppressor of cytokine signaling genes in myeloid leukemia and related proliferative disorders has also been further defined. Finally, positive signs have emerged in mice that exogenous delivery of SOCS3 may be of therapeutic value. SUMMARY: Suppressor of cytokine signaling proteins have pivotal roles in attenuating cytokine and toll-like receptor signaling in myeloid cells. Understanding how defective suppressor of cytokine signaling activity contributes to inflammatory and malignant disease promises to create significant new therapeutic opportunities. PMID- 17133095 TI - Multispecific myeloid defensins. AB - PURPOSE OF REVIEW: This review describes recent progress in our understanding of defensins and their contributions to innate immunity. Defensins are small, cysteine-rich endogenous antibiotic peptides. Human neutrophils contain large amounts of three alpha-defensins (HNP-1-HNP-3), and smaller amounts of a fourth, HNP-4. Monocytes and macrophages generally lack defensins, but they release messengers that induce the synthesis of beta-defensins in epithelial cells. RECENT FINDINGS: In addition to their antimicrobial and immunomodulatory effects, HNP-1-HNP-3 possess antiviral and toxin-neutralizing properties. Induction of beta-defensins in epithelial cells is mediated by cell-surface Toll-like receptors or cytoplasmic peptidoglycan receptors that can recognize pathogen associated molecules. Mutations in Nod2, a cytoplasmic peptidoglycan receptor, are associated with reduced levels of intestinal alpha-defensins and ileal Crohn's disease. Human defensin genes show marked copy-number polymorphism. High level constitutive expression of defensins may afford protection against HIV-1 and other defensin-sensitive pathogens. Theta-defensins (cyclic octadecapeptides found in nonhuman primates) have impressive antiviral and antitoxic properties. SUMMARY: The multiple properties of defensins contribute to human innate immunity against bacteria, bacterial toxins, and viruses. PMID- 17133096 TI - Severe congenital neutropenia: inheritance and pathophysiology. AB - PURPOSE OF REVIEW: Severe congenital neutropenia is a heterogeneous disorder of hematopoiesis characterized by a maturation arrest of granulopoiesis at the level of promyelocytes with peripheral blood absolute neutrophil counts below 0.5 x 10/l. In this review we summarize our current knowledge on inheritance and pathophysiolgy of congenital neutropenia. RECENT FINDINGS: There are two major subtypes of congenital neutropenia as judged by inheritance: autosomal dominant trait defined by neutrophil elastase mutations consisting of 60% of patients and autosomal recessive trait comprising approximately 30% of patients. This genetic heterogeneity suggests that several pathologic mechanisms may lead to the same phenotype due to downregulation of common myeloid transcription factors. Lymphoid enhancer-binding factor 1 is the most promising candidate, as its abrogation together with downregulation of lymphoid enhancer-binding factor 1 target genes is compatible with this phenotype. Congenital neutropenia is considered as a preleukemic syndrome, since after 10 years of observation the cumulative incidence for leukemia is 21%. Acquired granulocyte colony-stimulating factor receptor mutations are detected in approximately 80% of congenital neutropenia patients who developed acute myeloid leukemia. SUMMARY: Congenital neutropenia is a congenital disorder of hematopoiesis inherited by autosomal dominant or recessive traits. Downregulation of lymphoid enhancer-binding factor 1 is involved in the pathophysiology of all congenital neutropenia patients. Congenital neutropenia patients with acquired granulocyte colony-stimulating factor receptor mutations define a group with high risk for development of leukemia. PMID- 17133097 TI - Genetics, biology and clinical management of myeloid cell primary immune deficiencies: chronic granulomatous disease and leukocyte adhesion deficiency. AB - PURPOSE OF REVIEW: Chronic granulomatous disease and leukocyte adhesion deficiency are the major primary immune deficiencies affecting phagocytic blood cells. Major advances in clinical diagnosis and development of novel treatments for these disorders merit review. RECENT FINDINGS: Clinically beneficial gene therapy correction of X-linked chronic granulomatous disease in two adult patients was reported. Nonmyeloablative busulfan conditioning before administration of gene corrected autologous hematopoietic stem cells was likely an essential maneuver to achieve successful gene therapy. There is an increased association of autoimmune disorders with chronic granulomatous disease. Preimplantation genetic diagnosis of leukocyte adhesion deficiency-I led to the birth of a normal child. A canine model of leukocyte adhesion deficiency-I facilitated development of new nonmyeloablative hematopoietic stem cell transplant and gene therapy approaches to leukocyte adhesion deficiency. Nonmyeloablative transplantation may provide an effective, but less toxic approach for leukocyte adhesion deficiency in children. There have been advances in understanding the basis of leukocyte adhesion deficiency-II and III. SUMMARY: The most important subjects reviewed in this chapter include new advances in development of gene therapy for chronic granulomatous disease and leukocyte adhesion deficiency-I; transplantation for leukocyte adhesion deficiency-I; prenatal diagnosis of leukocyte adhesion deficiency-I; and association of autoimmune diseases with chronic granulomatous disease. PMID- 17133098 TI - Diagnosis and treatment of hypereosinophilic syndromes. AB - PURPOSE OF REVIEW: The aim of this article is to provide an update of causes of hypereosinophilia, including advances in knowledge of eosinophilic leukemia, and to outline an approach to investigation. We also aim to discuss in more detail the diagnosis and management of various hypereosinophilic syndromes including the clonal eosinophilias and those driven by abnormal cytokine-secreting T cells. RECENT FINDINGS: Our understanding of the causative genetic abnormalities in eosinophilic leukemia is increasing, as is the repertoire of techniques available to detect them. New treatments on the horizon include further tyrosine kinase inhibitors for use in eosinophilic leukemia, which should provide an alternative to imatinib for those patients who develop resistance. These may also prove useful for other hypereosinophilic syndromes without PDGFRA or PDGFRB rearrangements. Other new therapies including anti-IL5 monoclonal antibodies are proving beneficial for some patients, especially those with abnormal T-cell populations. SUMMARY: As our understanding of the various hypereosinophilic syndromes increases, and we are able to characterize many of the causative genetic lesions in the clonal eosinophilias, we are increasingly able to select appropriate therapy for an individual patient. New therapies based on this knowledge should serve to further improve the prognosis for many patients with hypereosinophilia. PMID- 17133099 TI - JAK-2 mutations and their relevance to myeloproliferative disease. AB - PURPOSE OF REVIEW: The identification of the JAK2V617F allele greatly improved our understanding of the molecular pathogenesis of myeloproliferative disorders. This review focuses on recent studies offering new genetic, biochemical, and functional insight into the role of JAK2V617F in the pathogenesis of these disorders. RECENT FINDINGS: JAK2V617F mutations are present in almost all patients with polycythemia vera, and in approximately half of those with essential thrombocytosis and myelofibrosis. JAK2V617F has constitutive tyrosine kinase activity, and is able to transform hematopoietic cells and activate JAK STAT signaling when co-expressed with homodimeric type I cytokine receptors. Murine bone marrow transplant experiments demonstrate that JAK2V617F is sufficient for the development of polycythemia vera in recipient mice. These results suggested that JAK-STAT pathway activation might contribute to the pathogenesis of JAK2-negative myeloproliferative disorders, and led to the discovery of an activating mutation in the thrombopoietin receptor in JAK2 negative myelofibrosis and essential thrombocytosis. SUMMARY: The discovery of the JAK2V617F allele represents an important advance in our understanding of the molecular pathogenesis of myeloproliferative disorders, though many questions remain regarding the role of a single allele in three clinically distinct disorders, the mechanism of activation of JAK2V617F, and the pathogenesis of JAK2 negative myeloproliferative disorders. PMID- 17133100 TI - Toll-like receptors and innate immunity in gut homeostasis and pathology. AB - PURPOSE OF REVIEW: Immune responses to commensal bacteria and resulting chronic inflammation are hallmarks of inflammatory bowel disease. Considerable evidence has accumulated to suggest that innate immune defense mechanisms interact with pro-inflammatory pathways and exacerbate or perhaps even initiate development of inflammatory bowel disease. RECENT FINDINGS: Recent cellular and mouse studies now show Toll-like receptor responses in intestinal epithelial cells are polarized and their activation by commensal bacteria plays an essential role in maintaining colonic homeostasis and controlling tolerance in the gut. Recent data also implicate signaling by Toll-like receptors and the common adaptor molecule MyD88 in intestinal epithelial homeostasis as well as the pathogenesis of inflammatory bowel disease, establishing a key link between the innate and adaptive immune defenses. SUMMARY: We review recent progresses in the understanding of the role of Toll-like receptors and other pattern recognition receptors in the host defense against gastrointestinal pathogens and maintenance of immune tolerance to commensal bacteria, highlighting areas that should provide fertile ground for future studies aimed at a more comprehensive understanding of the interplay between innate and adaptive immune defense mechanisms and inflammatory bowel disease. PMID- 17133101 TI - Intertwining of thrombosis and inflammation in atherosclerosis. AB - PURPOSE OF REVIEW: The aim of this article is to highlight the importance of thrombotic processes in the development and complications of atherosclerotic vascular disease. RECENT FINDINGS: Thrombin generated at sites of vascular inflammation activates major atheroma-associated cells including endothelial cells, platelets, smooth muscle cells, monocytes, and macrophages. Thrombin activated cells produce a plethora of inflammatory mediators, such as regulated upon activation normal T cell expressed presumed secreted, macrophage migration inhibitory factor, and CD40 ligand, that promote atherosclerotic lesion formation and atherothrombotic complications of vascular disease. Additionally, thrombin induced inflammatory mediators stimulate tissue factor procoagulant activity within atheroma to initiate a positive feedback loop where thrombin activation launches inflammatory signals that lead to further thrombin activation. Platelets, the main cellular effectors of the thrombotic system, also play a central role in the biology of atherosclerosis by producing inflammatory mediators and directing leukocyte incorporation into plaques through platelet mediated leukocyte adhesion. SUMMARY: New research has identified signaling pathways that intertwine thrombotic and inflammatory pathways with the development and progression of atherosclerosis. These signaling pathways contain positive feedback loops that propagate atherogenesis. Targeting molecular regulators at the interface of thrombosis and inflammation simultaneously may reduce thrombosis and inflammation, thus breaking pathological cycles that promote atherosclerosis and associated thrombotic complications. PMID- 17133102 TI - Bibliography. Current world literature. Myeloid biology. PMID- 17133104 TI - Psychiatric and cognitive aspects of primary hyperparathyroidism. AB - PURPOSE OF REVIEW: Clinical guidelines for the treatment of primary hyperparathyroidism have been established by the 2002 NIH workshop on asymptomatic primary hyperparathyroidism. The panel called for further study of the ill-defined psychiatric and cognitive changes often seen in patients with primary hyperparathyroidism. The present paper provides a rigorous, updated review of the most recent advances and studies that have measured health-related quality of life, neurocognitive and psychiatric changes, as well as neurophysiologic imaging in patients with primary hyperparathyroidism undergoing parathyroidectomy. RECENT FINDINGS: In studies conducted pre and postparathyroidectomy, six recent articles have described improvements in health related quality of life. Five studies included evaluations with validated psychiatric and cognitive tests in prospective case-control trials, and showed varied improvements in depression, memory and concentration after parathyroidectomy. Two studies evaluated in a preliminary fashion the brains of patients with primary hyperparathyroidism with functional imaging studies, showing regional cerebral blood flow changes and prefrontal cortical activation with sleep improvement in postsurgical patients. SUMMARY: The studies described in this paper underline the benefits of surgical treatment on nontraditional symptoms in patients with primary hyperparathyroidism, and open the door to the continued study of the endocrine effects of primary hyperparathyroidism on brain function. PMID- 17133105 TI - Controversies in the surveillance of patients with well differentiated thyroid cancer. AB - PURPOSE OF REVIEW: Thyroid cancer incidence is rising in the USA, likely due to increased surveillance. Over the past several years, new approaches have developed for the long-term monitoring of differentiated thyroid cancer patients. The most recent developments in this area, as well as interesting data on molecular genetics of thyroid cancer will be discussed. RECENT FINDINGS: The introduction of 18F-fluorodeoxyglucose positron emission tomography (18F-FDG PET) imaging in the follow-up of patients with differentiated thyroid cancer is an important development in the last few years. Recent research has helped improve understanding of how 18F-FDG PET can best be applied. There is now improved understanding of the use of serum thyroglobulin levels to predict future risk of recurrence of differentiated thyroid cancer. Guidelines from the American Thyroid Association for management of thyroid neoplasia were updated in 2006 for the first time in a decade, helping physicians navigate the published data and provide evidence-based care. Ongoing advances in thyroid cancer genetics may help predict aggressiveness of individual thyroid cancers. SUMMARY: Better tools are becoming available for physicians caring for thyroid cancer patients. Recent developments in thyroid cancer research will help physicians better anticipate a patient's future disease course and select appropriate surveillance testing. PMID- 17133106 TI - Molecular mechanisms involved in differentiated thyroid cancer invasion and metastasis. AB - PURPOSE OF REVIEW: The majority of patients with thyroid cancer have an excellent prognosis, however patients with extensive local invasion and distant metastasis frequently do not respond to standard treatments and have worsened prognosis. Understanding the specific mechanisms involved in thyroid cancer invasion and metastasis is critical in order to develop new treatments specifically targeted for these patients. RECENT FINDINGS: The genetic basis for thyroid cancer initiation and development is well characterized, with the majority of studies implicating activation of the RAS-RAF-ERK and PI3K/PDK1/Akt signaling pathways. Over the last several years, data from a concerted effort to define the pathways involved in invasion and metastasis suggest that reactivation of embryonic pathways involved in cell movement, to include epithelial to mesenchymal transition and collective cell migration, may be involved in cancer cell migration and invasion. The previously identified thyroid oncogenes, BRAF, RET/PTC and Ras, appear to be important regulators of this process. SUMMARY: The molecular mechanisms that control cell migration during embryological development, such as epithelial to mesenchymal transition, appear to be reactivated in invading thyroid cancer cells. Elucidation of the signal transduction networks and molecules that are involved in thyroid cancer invasion may lead to novel therapeutic targets. PMID- 17133107 TI - Medullary thyroid cancer: therapeutic targets and molecular markers. AB - PURPOSE OF REVIEW: The present review will provide an update of important studies in medullary thyroid cancer (MTC) with an emphasis on targeted preclinical and translational research studies published over the past 2 years. RECENT FINDINGS: Recent advances in the biology of MTC, particularly in RET proto-oncogene signaling, are now being translated into promising new therapies and biomarkers. Multifunction tyrosine kinase inhibitors that target RET, plus vascular endothelial growth factor receptors and additional kinases, are now being evaluated in Phase II clinical trials in MTC. Important unanswered questions include the optimal means for selecting high-risk patients, appropriate biomarkers for monitoring kinase inhibitor trials, and trial endpoints. Similar to ABL, epidermal growth factor receptors and other kinases, individual mutant RET forms have differential sensitivity to different inhibitors. In addition to RET, an old marker, calcitonin, has assumed increasing importance, but may not adequately reflect changes in tumor burden in RET inhibitor trials. A number of new therapeutic strategies are being developed that could be appropriate for the approximately 50% of patients who lack RET mutations in their tumors. SUMMARY: Progress is being made toward effective targeted MTC therapy. Patients with advanced, progressive MTC should be considered for enrollment in clinical trials. PMID- 17133108 TI - Carney complex: the first 20 years. AB - PURPOSE OF REVIEW: The purpose of this review is to comment on the current findings on Carney complex, a dominantly inherited disease and a unique multiple endocrine neoplasia syndrome. RECENT FINDINGS: Sequencing of the PRKAR1A gene in more than 150 kindreds has revealed a number of pathogenic mutations; in more than 90% of the cases, the sequence change was predicted to lead to a premature stop codon and, thus, mutant mRNAs were subject to nonsense-mediated mRNA decay. In Carney complex syndrome cells carrying these mutations, protein kinase A activity is irregularly stimulated by cAMP. Mutations that did not lead to a premature stop codon have also been described; these were also associated with abnormal protein kinase A activity. Animal models of the disease have been recently developed; they reproduced some of the stigmata of Carney complex syndrome but not all. Genetic testing of patients' family members has been introduced in recent years, leading to early detection and a better overall prognosis. SUMMARY: New treatments have yet to be applied; the elucidation of the molecular pathways regulated by PRKAR1A holds the promise of leading to molecularly designed therapies. PMID- 17133109 TI - Functioning and nonfunctioning neuroendocrine tumors of the pancreas. AB - PURPOSE OF REVIEW: Neuroendocrine tumors of the pancreas are a small subgroup of tumors characterized by a variety of biological behaviors. Recent changes in their classification should help better define the prognosis of this diverse group of tumors. With recent advances in diagnosis and staging, the treatment options for all neuroendocrine tumors have evolved. Presented here is a review of the current-day knowledge for neuroendocrine tumors of the pancreas. RECENT FINDINGS: A consensus by leading experts in the neuroendocrine tumors field has proposed an algorithm for the diagnosis, treatment and follow-up of these rare tumors. Surgical resection remains the first-line therapy. Alternative forms of cytoreduction such as radiofrequency ablation and embolization, have increased the ability of the surgeon to debulk these tumors, resulting in improved survival and better palliation. Contrary to adenocarcinoma of the pancreas, hormonal and biotherapy offer unique treatment strategies for these rare tumors. Very recent developments utilizing radionuclide therapy hold promise for not only palliation, but may prove to be a beneficial form of adjuvant therapy. SUMMARY: Presented here is a summary of the recent literature on the diagnosis and treatment of neuroendocrine tumors of the pancreas. PMID- 17133110 TI - Digital quantification of mutant DNA in cancer patients. AB - PURPOSE OF REVIEW: The accumulation of somatic mutations is the major driving force for tumorigenesis. These mutations uniquely differentiate tumor cells from their normal counterparts. Mutations within tumor cells and mutant DNA released by tumor cells into blood, lymph, stool, tissues and other bodily compartments can thereby be used for cancer detection. Here we discuss technologies available for the detection and quantification of mutant DNA in clinical samples and the value of such measurements for patient management. RECENT FINDINGS: Conventional mutation detection technologies are either qualitative or only roughly estimate the abundance of mutant DNA molecules. Recently-developed approaches, however, use single molecule counting to determine the genotype of each individual member of a DNA population, providing a more accurate and precise digital output. SUMMARY: In this review, we discuss the clinical utility of mutant DNA quantification in cancer patients in the context of recent technical advances made in digital mutation detection. PMID- 17133111 TI - Single nucleotide polymorphism array analysis of cancer. AB - PURPOSE OF REVIEW: Classifying tumors and identifying therapeutic targets requires a description of the genetic changes underlying cancer. Single nucleotide polymorphism (SNP) arrays provide a high-resolution platform for describing several types of genetic changes simultaneously. With the resolution of these arrays increasing exponentially, they are becoming increasingly powerful tools for describing the genetic events underlying cancer. RECENT FINDINGS: The ability to map loss of heterozygosity (LOH) and overall copy number variations using SNP arrays is known. Techniques have recently been developed to map LOH at high resolution in the absence of paired normal data. Copy number variations described by SNP array studies are now reaching resolutions enabling the identification of novel oncogenes and tumor suppressor genes. The ability to determine allele-specific copy number changes has only recently been described. Moreover, SNP arrays offer a high-throughput platform for large-scale association studies that are likely to lead to the identification of multiple germline variants that predispose to cancer. SUMMARY: SNP arrays are an ideal platform for identifying both somatic and germline genetic variants that lead to cancer. They provide a basis for DNA-based cancer classification and help to define the genes being modulated, improving understanding of cancer genesis and potential therapeutic targets. PMID- 17133112 TI - RNA interference technologies and their use in cancer research. AB - PURPOSE OF REVIEW: Recently, RNA interference has evolved into a powerful research tool to functionally characterize genes. Genome-wide RNA interference reagents can study the loss-of-function phenotypes of candidate genes in the context of various disease model systems. In this review, we discuss the data from the most recent studies using RNA interference reagents with a focus on RNA interference-based genomic screening as a tool to expand our knowledge about the molecular basis of cancer. RECENT FINDINGS: Tumorigenesis is the result of the progressive accumulation of mutations in genes controlling cell proliferation and death. Various genes carrying these alterations are known to be directly linked to tumor growth; however, how to translate this knowledge into effective chemotherapeutics, nontoxic to normal cells, is still a subject of intensive research. SUMMARY: Loss-of-function studies offer a potential for validation of known and unrecognized tumor-associated targets. RNA interference-mediated gene knockdown can be exploited to study the reprogrammed circuitry of genes, discover gene interactions restricted to cancer cells and identify mechanisms of chemoresistance in cancer cells. In addition, the simultaneous use of cancer drugs and RNA interference also provides a paradigm to develop strategies to inactivate essential genes promoting neoplastic growth. PMID- 17133113 TI - Ba/F3 cells and their use in kinase drug discovery. AB - PURPOSE OF REVIEW: Due to their ability to function as dominant oncogenes, protein kinases have become favored targets in the quest for 'molecularly targeted' cancer chemotherapeutics. The discovery of a large number of cancer associated mutations in the kinome, and the progress in developing specific small molecule kinase inhibitors has increased the need for accurate, reproducible, and efficient kinase activity-dependent cellular assay systems. RECENT FINDINGS: Ba/F3, a murine interleukin-3 dependent pro-B cell line is increasingly popular as a model system for assessing both the potency and downstream signaling of kinase oncogenes, and the ability of small-molecule kinase inhibitors to block kinase activity. Facilitated by their growth properties, Ba/F3 cells have recently been adapted to high-throughput assay formats for compound profiling. Further, several published approaches show promise in predicting resistance to small-molecule kinase inhibitors elicited by point mutations interfering with inhibitor binding. SUMMARY: Ba/F3 cells are an increasingly popular tool in kinase drug discovery. The ability to test the transforming capacity of newly identified kinase mutations, and to profile drug candidates and compound libraries in high-throughput fashion, combined with the use of Ba/F3 cells to predict clinical resistance will greatly facilitate developments in this field. PMID- 17133114 TI - Cancer stem cells and oncology therapeutics. AB - PURPOSE OF REVIEW: An overview of the latest developments in the cancer stem cells field and their potential use in the oncology drug discovery process. RECENT FINDINGS: Recent studies provided evidence of the existence of a subpopulation of cells within a variety of tumor types with a tumorigenic potential that is lacking in the rest of the cells within these tumors. There is mounting evidence that such cells exist in almost all tumor types. Work on the characterization of these cells suggests that deregulation of pathways responsible for stem cell self-renewal is a likely requirement for carcinogenesis and targeting such pathways might be curative. Progress has been made to develop more relevant in-vitro and in-vivo models that incorporate these findings. SUMMARY: Cancer stem cells have been identified in a variety of tumors. Characterization of these cells, determining how they originate and developing relevant assays is a work in progress. Incorporating these findings in the cancer drug discovery process might lead to better therapeutics. PMID- 17133115 TI - Bibliography. Current world literature. Endocrine tumours. PMID- 17133117 TI - Acute severe asthma: recent advances. AB - PURPOSE OF REVIEW: Acute severe asthma is challenging to the clinician both in terms of recognition and appropriate treatment. About 30% of these episodes need admission to the medical intensive care unit with a mortality of 8%. Relapse rates vary from 7 to 15% depending on how well the patient is managed. The purpose of this review is to discuss recent advances in identification of risk factors, pathophysiology and management of acute severe asthma. RECENT FINDINGS: Although the exact mechanism for acute severe asthma is unclear, some that are implicated include inflammation, airway remodeling and downregulation of beta receptors. None of the environmental factors have been clearly related to the development of near fatal attacks. Genetic polymorphisms have been associated with severe asthma. Lack of steroid responsiveness has been linked to severe asthma attacks. Chemokines and basement membrane changes characteristic of severe asthma are reported in a few studies. Lack of symptom perception in a certain group of patients with acute severe asthma leads to delayed interventions. Specific treatment modalities and ventilator management is reviewed. SUMMARY: Severe asthma is a phenotype of asthma with variable pathology and clinical presentation. Early recognition and timely intervention is needed to prevent significant mortality and morbidity. PMID- 17133118 TI - Bronchial asthma: the Indian scene. AB - PURPOSE OF REVIEW: Although asthma is a global disease, there are important differences in epidemiology, clinical spectrum and management practices in India. Some of these issues have been reviewed in this article. RECENT FINDINGS: The prevalence of 'ever asthma' was reported in 2.4% in a population study on 73,605 individuals conducted simultaneously at four major centres in India with the use of a single definition and uniform methodology employing a validated questionnaire. Of the several risk factors which were found to be significant, exposure to environmental tobacco smoke during childhood alone or both during childhood and adulthood was important in the development of and in increasing morbidity from asthma. Many other triggers and risk factors which include local aeroallergens and air pollutants have been identified. Allergic bronchopulmonary aspergillosis is an important cause of difficult to treat asthma and almost half of these patients receive antitubercular treatment at some stage. Simplified consensus guidelines based on international guidelines and local practices have been developed for use at the primary and secondary levels of healthcare. SUMMARY: The prevalence of asthma in India is somewhat similar to that seen in other Asian countries. Consensus management guidelines adapted from standard international guidelines adequately address the local concerns and issues. PMID- 17133119 TI - The connection between allergic rhinitis and bronchial asthma. AB - PURPOSE OF REVIEW: The relationship between allergic rhinitis and asthma has intrigued practicing physicians since the turn of the century. Investigations during the past two decades have led to an increased understanding of this relationship. RECENT FINDINGS: A growing body of evidence clearly demonstrates that patients with allergic rhinitis, in the absence of asthma, have distinct abnormalities of lower airway function, including alterations in physiology, histology and biochemistry. In addition, epidemiologic surveys have consistently shown allergic rhinitis as an independent risk factor for developing asthma, and that preexisting abnormalities in lung function may predispose to lower airway disease development. Clinical trials show that specific allergen immunotherapy for children and adults with seasonal allergic rhinitis reduces the risk of developing asthma. In patients with established asthma and concomitant allergic rhinitis, several medications have significant effects upon the upper and lower airways, including intranasal corticosteroids, oral antihistamines, and leukotriene receptor antagonists. SUMMARY: Our understanding of the natural history of allergic rhinitis could lead to improvements in early intervention, potentially preventing the progression of allergic rhinitis to asthma. Aggressive treatment of rhinitis in patients with concomitant asthma may enhance asthma outcomes and quality of life for all patients with these chronic diseases. PMID- 17133120 TI - Asthma phenotypes. AB - PURPOSE OF REVIEW: Asthma is a heterogeneous disorder presenting with many phenotypes. Precise phenotypic definition has eluded the medical research community for years, despite recognition of different disease subtypes. Improved phenotypic characterization and knowledge of underlying pathobiology is necessary for linkage of specific genotypes with clinical disease manifestations. RECENT FINDINGS: Phenotyping has been difficult because asthma is likely to be comprised of overlapping syndromes with varying origins and heterogeneous pathobiology. Currently, the field is too reliant on classification by trigger or symptoms. Since genotypic and phenotypic heterogeneity are inherent in asthma, patients presenting with different asthma phenotypes may need tailored therapies. Studies have begun to link genetics with disease mechanism and therapeutic response. As disease etiology, onset, progression and severity vary greatly among patients, however, the relative contribution of genetic factors may be difficult to ascertain. Definition of the full array of complex biological consequences of molecular target modulation is a prerequisite for therapies based on this concept. SUMMARY: The advent of targeted therapies for asthma and clinical trials based on phenotype and genotype have raised interest in more accurate description of asthma phenotypes. Therapies based on phenotypic and genotypic characteristics may be useful in asthma management. A variety of factors, however, must be addressed before such approaches become standard. PMID- 17133121 TI - Critical appraisal of antileukotriene use in asthma management. AB - PURPOSE OF REVIEW: Recognition of the importance of leukotrienes in the pathogenesis of asthma has led to the development of leukotriene modifiers, the first new class of drugs for asthma treatment to become available since the introduction of inhaled corticosteroids. Nevertheless, despite their widespread use in clinical practice, the role of leukotriene modifiers in the management of asthma remains controversial. In the present article the clinical applications of this class of drugs have been critically reviewed based on recent evidence. RECENT FINDINGS: In an effort to try and establish the proper place of antileukotrienes in the management of asthma, important systematic reviews have been carried out over these recent years in three critical areas: antileukotrienes as second choice first line agents after inhaled corticosteroids; antileukotrienes as add-on therapy to inhaled corticosteroids; add-on antileukotrienes versus long acting beta-agonists to patients not controlled by inhaled corticosteroids. In addition, novel and useful clinical targets for this class of drugs have been recently explored and include: patients with severe asthma; aspirin-intolerant asthma; asthmatic patients with allergic rhinitis. SUMMARY: Use of antileukotrienes is not recommended as first-line monotherapy in patients with asthma, except those who have aspirin intolerant asthma. Patients with concomitant allergic rhinitis may be a good target population for therapy with antileukotrienes. Addition of leukotriene modifiers to inhaled corticosteroids produces only a modest improvement in the clinical response, and is not greater to that of add-on long acting beta-agonists. The exact role of antileukotrienes in asthma management guidelines still continues to evolve. PMID- 17133122 TI - The role of chest radiography and computed tomography in the diagnosis and management of asthma. AB - PURPOSE OF REVIEW: The management of asthma is guided by clinical symptoms, physiological measurements, and response to therapy. Recent advances in computed tomography imaging promise to add a new dimension to our diagnostic armamentarium. Accurate representation of airway pathology, visualized by high resolution chest computed tomography scan, helps to improve the understanding of the pathophysiology of asthma. In addition, findings on computed tomography may help to guide therapies for asthma. As radiologists provide us with sophisticated modalities that may also have a bearing on treatment, clinicians should stay abreast of this evolving noninvasive technology. RECENT FINDINGS: This review focuses on the findings seen on computed tomography imaging as related to asthma. Airway wall thickness is discussed and how it relates to disease progression and pulmonary function test. In addition, indirect findings such as bronchial dilatation and mosaic attenuation, both consequences of air-trapping, are discussed. Other investigational tools, such as endobronchial ultrasound and positron emission tomography, are described. SUMMARY: New modalities in radiology hold promise to aid in the understanding and treatment of small-airway disease. Although still considered investigational modalities, research evidence is fast accumulating. It behooves the clinician to have a heightened awareness regarding further advancements in this field. PMID- 17133123 TI - Asthma: new developments concerning immune mechanisms, diagnosis and treatment. AB - PURPOSE OF REVIEW: This brief review discusses how recent research may modify our understanding of the immunology of asthma. Consideration is given to the possible impact that these observations may have upon diagnostic and therapeutic strategies. RECENT FINDINGS: New studies indicate that current conceptions regarding the balance between Th1 and Th2 systems may need modification. The relationship between infection and the development of asthma in children has proven to be much more complex than originally suggested by the 'hygiene hypothesis'. In addition, important genetic differences have been found in the response of asthmatic patients to therapeutic agents. SUMMARY: Greater insight into the mechanisms responsible for asthma and the introduction of new drugs will depend in part upon the development of reliable and simple methods for detecting airway inflammation. As the immunologic aspects of asthma are dissected, we can expect that many more potential targets for treatment will be discovered, but treatment may have to be individualized for genetic differences between different individuals. PMID- 17133124 TI - Cough and asthma. AB - PURPOSE OF REVIEW: The intention of this article is to discuss and place into perspective recent articles on cough and asthma. RECENT FINDINGS: Asthma continues to be a major diagnosis in most studies of cough. The first prospective study of sub-acute cough demonstrated an asthma incidence lower than that for chronic cough, a logical finding; upper airway cough syndrome often causes cough in the postinfectious state. The first prospective study of cough in infants suggested asthma to be a minor cause of cough in infants, but methodological flaws make the conclusions uncertain. Efforts to separate cough-variant asthma from classic asthma continue. One group has demonstrated that the maximal bronchoconstrictor response in cough-variant asthma is blunted when compared with classic asthma, a possible explanation for the absence of wheeze and dyspnea in cough-variant asthma. Another look at airway resistance showed a less rapid rate of rise in resistance in cough-variant asthma with increasing methacholine dosing than in classic asthma. On the biochemical front, a group has demonstrated differences in vascular endothelial growth factor, which may be the underpinnings of differences between cough-variant asthma and classic asthma. SUMMARY: Recent data suggest that cough-variant asthma is part of a continuum in the expression of asthma symptoms and in the asthmatic inflammatory response. PMID- 17133125 TI - Nocturnal asthma. AB - PURPOSE OF REVIEW: To examine recent advancements of the epidemiology, pathophysiology, and treatment of nocturnal asthma. RECENT FINDINGS: In a large cross-sectional survey, the prevalence of nocturnal symptoms was 60%. Forty-two percent of patients who completed the study did not declare nocturnal symptoms that were detected by physicians. A recent study including 47 individuals with nocturnal and nonnocturnal asthma showed that allele genotype Gly16 was significantly higher in the nocturnal asthma group (72%) compared with the nonnocturnal asthma group (54%). A randomized double-blind study of nocturnal asthma with esomeprazole improved the peak expiratory flow rate in individuals with both nocturnal asthma and gastroesophageal reflux disease. Of patients with nocturnal asthma and snoring, 48% had obstructive sleep apnea. After continuous positive airway pressure treatment, there was significant improvement of asthma nighttime symptom scores; however, no significant difference in lung functions was observed. SUMMARY: Nocturnal symptoms in asthma are frequently underdiagnosed. Obstructive sleep apnea may be prevalent in nocturnal asthma. The Gly16 polymorphism of the beta2-receptor is common in nocturnal asthma. Gastric suppression treatment in patients with both gastroesophageal reflux disease and nocturnal asthma improved the peak expiratory flow rate. Continuous positive airway pressure improves nocturnal symptoms but not lung function tests. PMID- 17133126 TI - Obesity and asthma: implications for treatment. AB - PURPOSE OF REVIEW: Epidemiological data as well as data from mouse models of asthma indicate a relationship between obesity and asthma. The purpose of this review is to evaluate recent data addressing this relationship and its biological basis, and to evaluate the implications of these data for treatment. RECENT FINDINGS: Obesity increases the prevalence, incidence, and possibly severity of asthma, while weight loss in the obese improves asthma outcomes. Obesity also influences asthma control and the response to standard asthma therapeutics. Moreover, obese mice exhibit innate airway hyperresponsiveness and increased responses to common asthma triggers. The biological basis for the relationship between obesity and asthma may be the result of common etiologies, comorbidities, effects of obesity on lung volume, or adipokines such as tumor necrosis factor alpha, leptin, and adiponectin. SUMMARY: Understanding the mechanistic basis for the relationship between obesity and asthma may lead to new therapeutic strategies for treatment of this susceptible population. PMID- 17133127 TI - Asthma and air quality. AB - PURPOSE OF REVIEW: There is evidence for an association between asthma and air pollutants, including ozone, NO2 and particulate matter. Since these pollutants are ubiquitous in the urban atmosphere and typically correlated with each other it has been difficult to ascertain the specific sources of air pollution responsible for the observed effects. Similarly, uncertainty in determining a causal agent, or multiple agents, has complicated efforts to identify the mechanisms involved in pollution-mediated asthma events and whether air pollution may cause asthma as well as exacerbate preexisting cases. RECENT FINDINGS: Numerous studies have examined specific sources of air pollution and their relationship to asthma. This review summarizes recent work conducted, specifically, on traffic pollution and presents results that elucidate several plausible biological mechanisms for the observed effects. Of note are studies linking susceptibility to several genetic polymorphisms. Together, these studies suggest that remaining uncertainties in the asthma-air pollution association may be addressed through enhanced assessment of both exposures and outcomes. SUMMARY: Air-pollution research is evolving rapidly; in the near future, clinicians and public health agencies may be able to use this new information to provide recommendations for asthmatics that go beyond only paying attention to the air pollution forecast. PMID- 17133128 TI - Allergic bronchopulmonary aspergillosis: a US perspective. AB - PURPOSE OF REVIEW: The present article is an update of allergic bronchopulmonary aspergillosis. Although a rare condition, allergic bronchopulmonary aspergillosis does affect a number of patients with asthma and cystic fibrosis. Prompt recognition and treatment of the disease is critical to improving patient outcomes. RECENT FINDINGS: There is currently much active research being performed in the area of allergic bronchopulmonary aspergillosis. Fascinating insights are being made into the pathophysiology and genetics of the disease. Additionally, research is ongoing on the use of recombinant Aspergillus allergens as an aid to the diagnosis of allergic bronchopulmonary aspergillosis. SUMMARY: These new insights into the genetics and pathophysiology of allergic bronchopulmonary aspergillosis and the development of these new diagnostic techniques could ultimately lead to improved patient treatment. These areas form an important basis for further research. PMID- 17133129 TI - Allergic bronchopulmonary aspergillosis: an Indian perspective. AB - PURPOSE OF REVIEW: Allergic bronchopulmonary aspergillosis is an immunologically mediated lung disease that is caused by hypersensitivity to antigens of the genus Aspergillus. This review summarizes the clinical presentation, radiologic profile, lung functions and immunologic studies on allergic bronchopulmonary aspergillosis from India. Data regarding Aspergillus sensitization in asthmatics are presented. The association of allergic bronchopulmonary aspergillosis with allergic Aspergillus sinusitis and aspergilloma is also highlighted. RECENT FINDINGS: Allergic bronchopulmonary aspergillosis is now an emerging disease in India. Sensitization to Aspergillus antigens is not uncommon in our patients with asthma. Although asthma commenced in these subjects in their early 20s, allergic bronchopulmonary aspergillosis was recognized more than a decade later. Allergic bronchopulmonary aspergillosis can also occur in patients without clinical asthma. Radiology is crucial to the diagnosis of allergic bronchopulmonary aspergillosis. The remarkable radiological similarity to pulmonary tuberculosis has important clinical implications in our country as patients with allergic bronchopulmonary aspergillosis often receive antituberculous therapy for a long time. Although oral corticosteroids still remain the cornerstone for management, itraconazole has emerged as an adjunct therapy in appropriate situations. Concomitant occurrence of allergic bronchopulmonary aspergillosis and allergic Aspergillus sinusitis is now being increasingly recognized. SUMMARY: All asthmatic subjects with a positive skin prick test to Aspergillus antigens must be evaluated for allergic bronchopulmonary aspergillosis and allergic Aspergillus sinusitis should be excluded. PMID- 17133130 TI - Current world literature. Asthma. PMID- 17133132 TI - Be aware of compassion fatigue. PMID- 17133133 TI - Are silver products safe and effective for chronic wound management? PMID- 17133134 TI - Analyzing focus group interviews. PMID- 17133135 TI - Relative delivery efficiency and convenience of spray and ointment formulations of papain/urea/chlorophyllin enzymatic wound therapies. AB - OBJECTIVE: To determine the relative delivery efficiency and user preference for spray and ointment formulations containing papain, urea, and chlorophyllin copper complex sodium (papain/urea/chlorophyllin copper complex). METHODS: Participants applied both a spray and 3 ointment formulations of papain/urea/chlorophyllin copper complex to identical wound models. The average amount of product used per application was determined by weighing the bottle or tube before the study and after completion of the study. Participants were also asked to fill out a questionnaire regarding their preferences for the two formulations. RESULTS: The amount of product used per wound application was 30% less with the spray formulation; resulting in 29% more wound applications per container. Over 80% of the study participants found the spray easier and quicker to use than the ointments. CONCLUSIONS: The spray formulation of papain/urea/chlorophyllin copper complex resulted in less product use per wound application than did 3 different ointment formulations. Participants expressed a favorable impression of practical benefits of the spray formulation including a reduced risk of wound contamination. PMID- 17133136 TI - When trauma means a stoma. AB - Trauma is a leading cause of death and disability. When traumatic injuries require ostomy surgery, the wound, ostomy, and continence nurse acts as a crucial part of the trauma team. This literature review describes mechanisms of injury associated with creation of a stoma, key aspects of wound, ostomy, and continence nursing care in trauma populations and presents suggestions for future research. PMID- 17133138 TI - Quality of life and coping of persons with temporary and permanent stomas. AB - PURPOSE: To evaluate and compare the use of coping strategies and quality of life (QoL) among individuals with temporary and permanent stomas. DESIGN: Descriptive, cross-sectional. METHODS: After consideration of ethical issues, 42 subjects with temporary stomas and 72 subjects with permanent stomas were interviewed. The Coping Strategies Inventory of Folkman and Lazarus, and Ferrans and Powers QoL Index, both validated for the Brazilian culture, were administered. RESULTS: Subjects from both groups used all coping strategies queried in the Coping Strategies Index and QoL index, but significant differences (P<0.05) were found in use of confrontive, escape-avoidance, and positive reappraisal factors, which were more frequently employed by patients with temporary stoma. QoL scores did not differ between the groups. Significant correlations (P<.001) among subjects with temporary stomas were observed between the family QoL subscale and (a) distancing, (b) self-control, (c) accepting responsibility, (d) escape-avoidance, and (e) positive reappraisal coping factors. Significant correlations were also found between the psychological/spiritual QoL subscale and positive reappraisal factor; between the health/functioning QoL subscale and planful problem solving and positive reappraisal for patients with temporary stomas. Among subjects with permanent stomas, only significant correlations existed between the psychological/spiritual QoL subscale and self-control and social support coping factors. CONCLUSION: Individuals with stomas tend to show positive QoL scores. However, different coping strategies were used by persons with temporary versus permanent stomas. PMID- 17133139 TI - Development of methods for skin barrier peeling tests. AB - OBJECTIVE: We sought to develop a more effective method to evaluate the adhesive properties of skin barriers. DESIGN: The experimental design used was based on 3 principles: partial control, randomization, and repetition. Using these principles, the 180-degree peeling tests were conducted as specified in a standardized methodology (JIS Z0297) to the extent possible. However, the use of a stainless steel plate as a proxy for skin barrier application may result in the stretching and breaking of the skin barrier, making it impossible to obtain suitable measurements. INSTRUMENTS: Tests were conducted in constant temperature/ humidity chambers using a Tensilon Automatic Elongation Tester, where a sample was fixed on the side of a sample immobilization device, a sturdy metal (aluminum) box from which the air in the box was drawn off with a vacuum pump. METHODS: A fluorocarbon polymer film was applied to the adhesive surface of a sample skin barrier. The film was peeled off in the volte-face (180-degree) direction in order to measure adhesive strengths. RESULTS: The films exhibit such properties as (a) ease of removal from the adhesive surface, (b) no resistance to a 180-degree fold back due to the thinness and flexibility of the material, and (c) tolerance of elongation. The adhesive properties of skin barriers were measured by peeling the fluorocarbon polymers in a 180-degree direction. Twelve specimen skin barrier products were selected for measurement, providing results with satisfactory reproducibility. Results based on the conventional stainless steel plate-based testing method acted as a control. CONCLUSION: The newly developed testing method enables chronological measurement results for skin barriers applied to fluorocarbon polymer films after 24 hours, 48 hours, and longer period. PMID- 17133140 TI - The experience of flatus incontinence from a bowel ostomy: a hermeneutic phenomenology. AB - OBJECTIVE: To interpret and present possible meanings in the stories of people with bowel ostomies about their experience of impact of flatus incontinence on their life and being. DESIGN: Hermeneutic phenomenology guided by a Gadamerian perspective. SETTING AND SUBJECTS: Six people with a bowel ostomy were recruited from a city in Australia. METHODS: In-depth, nonstructured interviews generated rich text. Interviews were videotaped. A variety of interpretive, hermeneutic techniques were applied for text interpretation RESULTS: Nine existential themes of meaning emerged: I am undignified, I am a secret, I am always with gas, I am not myself alone, I am without choice, I am a seeker of control, I am the smell, I am not normal, and I am living a life-sort of. Through symbiotic interpretation, writing, and re-writing, themes were encompassed in a short story: a creative synthesis of actual events and interpreted understandings for ostomates about possible meanings of experiencing flatus incontinence. CONCLUSION: Flatus incontinence for people with bowel ostomies can be quite discommoding and impact on their interactions, self-image, sexuality, social activity, and psychological well-being. Nurses need to understand this for empathetic interaction, patient assessment, intervention selection, research planning, and pertinent education. PMID- 17133141 TI - Perioperative factors that affect long-term adjustment to an incontinent ostomy. AB - PURPOSE: We sought to identify perioperative factors associated with long-term adjustment to an incontinent ostomy. METHODOLOGY: The Ostomy Adjustment Scale (OAS), a demographic form, and a $5 gift certificate were mailed to 200 ostomates. Associations between items from the Survey of Perioperative Factors of Ostomy Adjustment, demographic forms, and total OAS scores were assessed using bivariate and multiple linear regression analysis. RESULTS: The survey response rate was 74%. About half (70/146, 49%) of the respondents were women, 99% (144/145) were Caucasian, and 74% (109/146) were married. Their mean age was 65+/ 14 years. Fifty-six percent (79/141) of the respondents had an ileostomy, 28% (39/141) had a colostomy, and 15% (21/141) had a urostomy. The mean OAS score for all the respondents was 159.5+/-26.2, indicating a good overall adjustment to an incontinent ostomy. In bivariate analyses, the factor associated with a lower OAS score indicating poorer adjustment was distress related to affording and obtaining ostomy supplies. The more helpful the ostomates found the preoperative education provided by a wound, ostomy, and continence specialist nurse, the better was their adjustment as indicated by a higher OAS score. Having ongoing/recurrent illness and having an ostomy created by a specialty surgeon were also associated with better adjustment. When multiple factors were considered in a stepwise regression analysis, adjustment was lowered by distress about obtaining ostomy supplies. Adjustment was improved when preoperative education provided by a wound, ostomy, and continence specialist nurse was considered helpful for the ongoing/ recurrent illness. CONCLUSION: Many ostomates have a good long-term adjustment to ostomy surgery and value education provided by a wound, ostomy, and continence nurse. Perioperative patient education and addressing patient distress over obtaining supplies are areas that can benefit from wound, ostomy, and continence nurse intervention. PMID- 17133142 TI - Privacy for defecation and fecal incontinence in older adults. AB - INTRODUCTION: Privacy during defecation is important to individuals and society at large and it has not been studied in older people with fecal incontinence. METHODS: One hundred twenty adults aged 65 years and with fecal incontinence who were either living in their own homes or in a nursing home or receiving care in an acute or rehabilitation elderly care ward were surveyed with a questionnaire that included questions on privacy during defecation. RESULTS: Privacy while defecating was often least achieved in the patients with fecal incontinence living in nursing homes (NH) (n=7, 23%) but usually was achieved in those living at home (H) (n=28, 93%) and by some being cared for in rehabilitation wards (R) (n=16, 53%) or in acute wards (AC) (n=15, 50%; P<.001). Very few participants with fecal incontinence were aware of leakage (NH: n=3, 10%; R: n=9, 30%; AC: n=9, 30%; H: n=15, 50%), able to clean themselves (NH: n=0, 0%; R: n=2, 7%; AC: n=5, 17%; H: n=24, 80%) or had access to patient information leaflets about fecal incontinence (NH: n=3, 10%; R: n=4, 13%; AC: n=1, 3%; H: n=16, 53%). CONCLUSION: Older people, especially those who are dependent, lack privacy during bowel movements. They are usually unaware of being incontinent of stool and are unable to clean themselves afterwards. Access to information about fecal incontinence is poor. PMID- 17133143 TI - Wet colostomy and peristomal skin breakdown. PMID- 17133144 TI - Fitting in or standing out: nursing workgroup diversity and unit-level outcomes. AB - BACKGROUND: Despite growing diversity in the nursing workforce, there has been limited investigation of the implications of diversity to the performance of nursing workgroups. OBJECTIVES: The aim of this study was to examine relationships among two categories of nursing workgroup diversity (highly visible and job related), intervening workgroup processes (workgroup cohesion and initiative), and workgroup performance (affective and cognitive). METHODS: A descriptive design with path analysis was used to test two models derived from Pelled's intervening process theory. Registered nurse (RN) and patient data from 222 nursing units at 126 hospitals, collected as part of the Outcomes Research in Nursing Administration Project (ORNA-II), were used. Registered nurses provided demographic data used to estimate workgroup diversity (age, race, unit tenure, educational background, and RN experience) and completed questionnaires measuring workgroup cohesion and initiative. Patients completed a satisfaction survey, and these data, along with unit-level patient falls and medication errors, were used to measure workgroup performance. RESULTS: Our findings failed to support the intervening process theory. No direct effects between diversity and intervening process variables were found. Cohesion and initiative did not mediate the relationships between diversity and workgroup performance. However, workgroup cohesion predicted greater patient satisfaction (p < .01) and higher levels of met expectations for symptom management (p < .01), whereas workgroup initiative predicted fewer patient falls (p < .01). DISCUSSION: Diversity may have little, if any, effect on nursing workgroup performance. Cohesion and initiative warrant further research as possible mediators of the relationship between patient outcomes and variables like nurse staffing, experience, and expertise. PMID- 17133145 TI - The anniversary of birth trauma: failure to rescue. AB - BACKGROUND: The reported prevalence of posttraumatic stress disorder secondary to birth trauma ranges from 1.5% to 5.6%. Serious ramifications of birth trauma are beginning to be recognized, such as impaired mother-infant interaction. OBJECTIVE: The aim of this study was to determine the essence of mothers' experiences regarding the anniversary of their birth trauma. METHODS: Colaizzi's method of phenomenology was used to guide the study. Participants were recruited via the Internet through a charitable trust located in New Zealand called Trauma and Birth Stress. Thirty-seven women sent attachment stories describing their experiences of the anniversary of their traumatic childbirths. RESULTS: Four themes revealed the essence of women's experiences of the anniversary of their birth trauma: (a) The prologue: An agonizing time; (b) The actual day: A celebration of a birthday or the torment of an anniversary; (c) The epilogue: A fragile state; and (d) Subsequent anniversaries: For better or worse. DISCUSSION: Based on the findings of this study on the anniversary of traumatic childbirths, the time seems right to broaden the use of the term failure to rescue to these childbearing women. Not only clinicians but also family and friends failed to rescue mothers during the period surrounding the anniversary of their birth trauma. PMID- 17133146 TI - Ethical conflict in nurse practitioners and physician assistants in managed care. AB - BACKGROUND: More patients are receiving healthcare services from nurse practitioners (NPs) and physician assistants (PAs). These providers are likely to be confronted with a variety of ethical issues as they balance quality care with their patients' rising cost concerns. However, very little is known about the ethical conflicts and causes of these conflicts experienced by these clinicians in their daily practice. OBJECTIVE: To identify ethical concerns and conflicts NPs and PAs encounter related to managed care in the delivery of primary care to patients and the factors that influence ethical conflict. METHODS: A cross sectional self-administered mailed questionnaire was sent to 3,900 NPs and PAs randomly selected from primary care and primary care subspecialties in the United States (adjusted response rate, 50.6%). Respondents were surveyed on ethical issues and concerns, ethics preparedness, and ethical conflict. Bivariate and multiple linear regression analyses were used to evaluate predictors of ethical conflict. RESULTS: Insurance constraints were reported to have interfered with the ability to provide quality patient care by 72% of respondents, with 55.3% reporting daily to weekly interferences. Nearly half of respondents (47%) have been asked by a patient to mislead insurers to assist them in receiving care. A perceived obligation to advocate for patients, even if it means exaggerating the severity of a patient's condition, was the single most significant predictor of ethical conflict, explaining 25% of the variance. DISCUSSION: NPs and PAs are experiencing ethical conflict often associated with their perceived professional obligations to advocate for patients. Being well-prepared in ethics and having sufficient professional independence help clinicians balance the ethical complexities and demands of meeting patients' needs within a constrained healthcare system. PMID- 17133147 TI - A randomized controlled trial of a smoking cessation intervention for pregnant adolescents. AB - BACKGROUND: The smoking prevalence rate among pregnant adolescents has been estimated at 59-62%, and 60-80% of these adolescents continue to smoke throughout their pregnancies. OBJECTIVES: The aim of this study was to evaluate the short- and long-term effects of smoking cessation strategies tailored to the pregnant adolescent to attain and maintain abstinence. The specific aim was to examine differences in short- and long-term smoking behaviors among three groups: Teen FreshStart (TFS), Teen FreshStart Plus Buddy (TFS-B), and Usual Care (UC) control. METHODS: In this randomized controlled intervention study, a 3-group (TFS, TFS-B, and UC) by 3-occasion (baseline, 8 weeks postrandomization, and 1 year following study entry) design was used. The study included 142 pregnant adolescents who were aged 14 to 19 years. Both self-reported smoking status collected on the Smoking History Questionnaire and saliva cotinine levels were used to identify smoking behaviors. RESULTS: There were no significant differences among the three treatment groups at baseline in terms of the racial distribution, age, gestational age, age of menses initiation, number in family household, number of family members who smoked, or tobacco use. A significant difference between the UC group and the TFS-B group (p = .010) was seen in smoking behaviors measured 8 weeks following treatment initiation. At 1 year following study entry, however, there were no differences between the groups in smoking behaviors. DISCUSSION: The TFS-B intervention was more effective in attaining short-term smoking cessation in the pregnant adolescent than TFS or UC. Findings suggest that the peer-enhanced programming had a limited effect but could not sustain the participant beyond postpartum (1 year following study entry). Future studies should include relapse prevention to sustain smoking abstinence into the postpartum period. PMID- 17133148 TI - Psychometric testing of the social support survey on a Taiwanese sample. AB - BACKGROUND: The Medical Outcomes Study Social Support Survey (MOS-SS) is a multidimensional, self-administered instrument used to assess various functional dimensions of social support. Less is known regarding its value for a Chinese population. OBJECTIVE: The aim of this study was to assess the psychometric properties of the MOS-SS on an adult sample in Taiwan. METHODS: A total of 265 adult family caregivers of patients with cancer from four different hospitals in the north, middle, and south regions of Taiwan completed the MOS-SS. RESULTS: In the original five-factor model, unsatisfactory item discriminant validity was found in almost half of the items; the item-own subscale correlation was lower than the item-other subscale correlation. A two-factor model accounting for 68.98% of the variance was found using exploratory factor analysis. The first factor (emotional support) accounted for 62.28% of the total variance, whereas the second factor (tangible support) accounted for 6.7%; the interfactor correlation was .71. The two-factor model seemed to have satisfactory reliability and validity and better discrimination between different subscales than did the original five-factor model. DISCUSSION: Good reliability and validity were demonstrated in the MOS-SS when applied to an adult sample in Taiwan. A two factor model, instead of a five-factor model as found in the Western countries, was found for this sample. Confirmation of the two-factor model and exploration of the two-factor model in related concepts are suggested for future studies. PMID- 17133149 TI - C-reactive protein in nipple aspirate fluid: relation to women's health factors. AB - BACKGROUND: Proteins expressed in nipple aspirate fluid (NAF) have been associated with breast cancer risk and may serve to detect inflammatory or premalignant states. Obesity, hyperlipidemia, and metabolic syndrome are associated with increased rates of breast cancer and are systemic markers of chronic inflammation. OBJECTIVE: The aim of this study was to determine if one NAF protein, C-reactive protein (CRP), associated elsewhere in the human with cancer, relates to reproductive, nutrition, and body composition and activity factors. METHODS: Women (n = 59) aged 30 to 64 years from the midwestern United States and Kenya were evaluated regarding the relation between reproductive (age, pregnancy, and breast-feeding indices), nutrition (serum lipids, serum, or NAF carotenoid), body composition and activity (activity, waist-hip ratio, fat percentage (% fat), and body mass index [BMI]) factors and CRP in NAF. RESULTS: Age at first pregnancy (p < .05), gravidity (p < .05), wean time from breast feeding last baby (p < .05), serum triglycerides (p = .01), % fat (p < .05), and BMI (p < .05) were related significantly to CRP levels in NAF. A model derived from selective women's reproductive, nutritional, and body composition and activity factors significantly (p < .05) accounted for the variance in breast microenvironment inflammation as measured by CRP in NAFs. DISCUSSION: Detecting CRP in NAF may indicate local mammary inflammation, which has been associated with carcinogenesis. Specific NAF risk modeling that addresses reproductive, nutritional, and body composition and activity factors may be used for further advances in the prevention of breast cancer and the early detection and treatment of the disease. PMID- 17133150 TI - Correlates of caregiver burden after coronary artery bypass surgery. AB - BACKGROUND: Coronary artery bypass (CAB) patients often rely on spouses for care and assistance during recovery after surgery. Caregiving can be stressful and meaningful depending upon the interplay of many factors not completely understood. These factors may affect the spouse caregiver's health-related quality of life (HRQL), which may impact ability to care for the CAB patient. OBJECTIVE: To investigate patient-spouse caregiver relationship and role variables associated with caregiver burden during the first year after CAB surgery. METHODS: Using a cross-sectional design, a convenience sample of CAB spouses was recruited at 3, 6, or 12 months. Spouses (n = 166) completed a survey that included perceptions of patient health status, caregiver burden and other caregiving variables, and caregiver HRQL. RESULTS: Higher burden scores were associated with patient's gender (female), poorer patient health status, lower caregiver mental HRQL, increased personal gain, and increased caregiver competence. These correlates explained 38% of the variance in caregiver burden. DISCUSSION: Only poorer patient health status and lower caregiver mental HRQL were supported by previous research in this population. Positive relationships between burden and caregiver competence and personal gain may be a reflection that spouse caregivers were invested and working hard. They felt satisfied from enhancement of self but were burdened from their caregiving role, providing support for a previously described two-domain caregiving model. Caregiver screening is essential to identify spouses at high risk for negative outcomes. Longitudinal research is needed to identify the correlates and predictors most likely to influence burden and caregiver gain over time, and to more fully understand caregiving in the CAB population. PMID- 17133151 TI - Mediators of the relationship between social support and positive health practices in pregnant women. AB - BACKGROUND: Literature suggests that there is a need to study a broadly conceived variable of positive health practices in pregnant women and to examine theoretically related variables that help to explain positive health practices in pregnant women. OBJECTIVES: The aims of this study were to examine the association between social support and positive health practices in pregnant women and to further test this relationship by examining two mediational variables: perceived health status and optimism. METHOD: A convenience sample of 152 pregnant women attending childbirth education classes completed the four study instruments and a demographic data sheet. RESULTS: Social support was related positively to positive health practices; this relationship was basic to both mediational models. In the first mediational model, social support was related positively to perceived health status, and perceived health status was related positively to positive health practices, but perceived health status was not a complete mediator of the social support-positive health practices relationship, as determined by regression analyses. In the second mediational model, social support was related positively to optimism, and optimism was related positively to positive health practices, but optimism was not a complete mediator of the social support-positive health practices relationship, as determined by regression analyses. DISCUSSION: Positive health practices in pregnant women are related positively to social support, perceived health status, and optimism. In terms of theory building, neither perceived health status nor optimism help to explain the social support-positive health practices relationship. Replication of this study with a more diverse sample is one recommendation for future study. PMID- 17133153 TI - The role of measles elimination in development of a national immunization program. AB - The U.S. Immunization Program has been one of the most successful efforts in preventive medicine. Since its beginning with passage of the Vaccination Assistance Act in 1962, polio, measles and rubella have been eliminated and many other vaccine-preventable diseases are at record or near record lows. In 1966, 3 years after licensure of the first measles vaccines, the Centers for Disease Control and Prevention began an effort to eliminate measles within the United States, an on-and-off effort that was to last more than 30 years. With measles elimination as the primary driver, fundamental components of today's immunization program were built that affected not only measles, but all of the vaccines and vaccine-preventable diseases of childhood. Some of the major contributions were the enactment and enforcement of immunization requirements for school attendance in all 50 states, enactment of an entitlement program for vaccine purchase, the Vaccines for Children Program, support for health services research to determine reasons for nonimmunization and interventions to improve coverage, development of standards for immunization practices and the measurement system for immunization coverage in all 50 states and 28 major urban areas. Key lessons have been: (1) the program must rest on a sound base of vaccine science and health services science; (2) having a limited number of measurable goals allows program focus, but consider strategies that have crosscutting impact; (3) accountability is critical to program performance at all levels-state, local and individual practice; and (4) establishing and maintaining political support is essential. PMID- 17133154 TI - An open-label, double tympanocentesis study of levofloxacin therapy in children with, or at high risk for, recurrent or persistent acute otitis media. AB - BACKGROUND: Levofloxacin has excellent activity against common respiratory pathogens and therefore is likely to be effective in treating children with persistent or recurrent otitis media. OBJECTIVE: The objective of this study was to assess the efficacy and safety of levofloxacin treatment in the eradication of bacterial pathogens from the middle ear fluid (MEF) of children with, or at high risk for, persistent or recurrent otitis media. METHODS: An open-label multicenter trial was conducted that involved tympanocentesis at entry and selectively 3 to 5 days after starting levofloxacin (10 mg/kg twice a day for 10 days). RESULTS: : Two hundred five children (80% < or =2 years) were enrolled. One child did not have a confirmed diagnosis of acute otitis media and did not return for follow-up visits. Of the remaining 204 children, 94 (46%) had bilateral infection and 63 (31%) were receiving antimicrobials immediately before entry. One hundred five isolates of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Streptococcus. pyogenes were recovered in pure or mixed cultures. All isolates were susceptible to levofloxacin. During treatment bacterial eradication from MEF occurred in 88% (78 of 89) of bacteriologically evaluable patients, including 90% (65 of 72) of children < or =24 months of age. Bacteria initially isolated from MEF were eradicated in 31 of 37 (84%) children infected with S. pneumoniae and in 54 of 54 (100%) children infected with H. influenzae. Overall, clinical success rate after therapy was 94% for the total study population and 92% for the bacteriologically evaluable population. Levofloxacin was well tolerated. Vomiting (4%) was the most common treatment-limiting adverse event. CONCLUSIONS: Levofloxacin was safe and effective in treating and eradicating common bacterial pathogens from MEF in children with, or at risk for, recurrent or persistent otitis media. PMID- 17133155 TI - The pharmacokinetics and safety of micafungin, a novel echinocandin, in premature infants. AB - BACKGROUND: : Candidal fungal infection rates in neonates are increasing and are a significant cause of mortality, especially in low birth weight infants. Micafungin is an echinocandin that works by inhibiting 1,3-beta-D-glucan synthase, an enzyme responsible for fungal cell wall synthesis. The objective of this study was to determine the safety and pharmacokinetics of micafungin in premature infants. METHODS: : This was a phase I, single-dose, multicenter, open label, sequential-dose trial of intravenous micafungin investigating 3 doses (0.75 mg/kg, 1.5 mg/kg and 3.0 mg/kg) in 18 premature infants weighing >1000 g (n = 6 in each dosage group). A further 5 infants (500-1000 g) were enrolled in the 0.75 mg/kg dosage group only. RESULTS: : The mean +/- standard deviation gestational age in the >1000 g dosage group was 26.4 +/- 2.4 weeks and, on entry, patients had one or more of a variety of underlying conditions, including sepsis, pneumonia and other infections caused by Candida or other species. Micafungin pharmacokinetics in preterm infants appears linear. However, premature infants >1000 g on average displayed a shorter half-life (8 hours) and a more rapid rate of clearance (approximately 39 mL/h per kg) compared with published data in older children and adults. All doses of micafungin were well tolerated and no serious drug-related adverse events were observed. CONCLUSIONS: : Single doses of micafungin, ranging up to 3.0 mg/kg, appear well tolerated in premature infants weighing >1000 g. The drug's elimination half-life and total plasma clearance in preterm infants appear dissimilar to published values for these parameters in older children and adults. The reason(s) for this apparent difference remain to be investigated. PMID- 17133156 TI - Decline in respiratory syncytial virus hospitalizations in a region with high hospitalization rates and prolonged season. AB - BACKGROUND: During 1993 to 1996, Alaska Native infants <1 year of age from the Yukon Kuskokwim (YK) Delta in Alaska experienced a respiratory syncytial virus (RSV) hospitalization rate 5 times the U.S. general infant population rate. We describe trends in lower respiratory tract infection (LRTI) and RSV hospitalizations in YK children from 1994 to 2004. METHODS: We abstracted hospital dates, RSV test results and clinical information from the hospital records for YK children <3 years of age hospitalized between July 1994 and June 2004. RESULTS: : The RSV hospitalization rate in YK Delta children <1 year of age decreased from 178 per 1000 infants per year (1994-1997) to 104 per 1000 infants per year (2001-2004) (P < 0.001), and the RSV hospitalization rate for premature infants decreased from 317 to 123 per 1000 infants per year (P < 0.001). The risk reduction for RSV hospitalization was greater in premature (relative risk, 0.39) than in term infants (relative risk, 0.60; P = 0.04). The rate of non-RSV LRTI hospitalizations increased from 153 to 215 per 1000 infants per year (P < 0.001). The median RSV season length was 30.5 weeks. Pneumonia was diagnosed in more than half of RSV admissions. CONCLUSIONS: In YK infants, the RSV hospitalization rate decreased by one-third between 1994 and 2004; however, the overall LRTI hospitalization rate did not change. The median RSV season was twice as long as for the U.S. population. Palivizumab prophylaxis may be responsible for the larger decrease in the RSV hospitalization rate among premature infants; however, the 2001-2004 RSV hospitalization rate among YK infants remained 3 times higher than the U.S. infant rate. PMID- 17133157 TI - A case-control study to determine risk factors for hospitalization for rotavirus gastroenteritis in U.S. children. AB - OBJECTIVE: The objective of this case-control study nested within a surveillance study conducted at 3 hospitals (Cincinnati Children's Hospital Medical Center, Cincinnati, OH; Children's Hospital of New Orleans, New Orleans, LA; and Hasbro Children's Hospital, Providence, RI) was to identify risk factors for rotavirus gastroenteritis requiring hospitalization. PATIENTS: Cases were children < or =59 months of age who were admitted with acute gastroenteritis (AGE) and found to have rotavirus infection. Controls were selected from a birth certificate registry (Cincinnati and Providence) or a registry of patients from a large practice consortium in 11 locations (New Orleans). RESULTS: Three hundred forty nine rotavirus-infected cases and 1242 control subjects were enrolled. Breast feeding was protective against hospitalization for rotavirus AGE for infants <6 months of age. (odds ratio [OR], 5.1; 95% confidence interval [CI], 1.2-13.2). Low-birth-weight (<2500 g) infants had increased risk for hospitalization even beyond the first few months of life (OR, 2.8; 95% CI, 1.6-5.0). Children in child care were more likely to be hospitalized for rotavirus AGE than those cared for at home, particularly those > or =24 months of age (OR, 3.0; 95% CI, 1.8-5.3). Other characteristics associated with rotavirus AGE hospitalization were children <24 months of age covered by Medicaid or without insurance (OR, 2.1; 95% CI, 1.4 3.2) and having another child in the house <24 months of age (OR, 1.6; 95% CI, 1.1-2.3). The data suggest that maternal age <25 years (OR, 1.4; 95% CI, 1.0-2.0) and a mother with less than a high school education (OR, 1.5; 95% CI, 1.0-2.3) may also increase risk of rotavirus hospitalization. CONCLUSION: There are socioeconomic and environmental factors and aspects of the child's medical and dietary history that identify children at risk for hospitalization with rotavirus AGE. PMID- 17133158 TI - Case-control study of vaccination history in relation to pertussis risk during an outbreak among school students. AB - BACKGROUND: Between September and December 2003, an outbreak of pertussis occurred in Cass County, MO, mostly among adolescent school children. METHODS: We conducted a 1:2 matched case-control study among school children and used conditional logistic regression to evaluate risk factors for pertussis, including the total number of vaccine doses received, age at administration of each dose of vaccine and the type of vaccine (whole cell or acellular). RESULTS: Of all 127 pertussis cases reported in this outbreak, the majority were adolescents (10-19 years of age, 50%) and adults (20 years or older, 22%); only 10% were infants and children less than 5 years of age. Because the focus of our investigation was on school-aged children, we enrolled 237 students (79 cases and 158 controls) in our study. Students missing at least one dose of the vaccine had higher risk for pertussis than those who received all 5 doses (odds ratio [OR], 2.36; 95% confidence interval [CI], 1.17-4.77). Early administration of the fifth dose of the vaccine at age 4 years was significantly associated with risk for pertussis compared with vaccination at age 5 years (adjusted OR, 2.45; 95% CI, 1.16-5.16). A short time interval (<36 months) between the fourth and fifth doses of the vaccine also tended to increase the risk for pertussis, although this association was not statistically significant. The type of vaccine was not a significant risk factor. CONCLUSION: Administering all 5 doses of pertussis vaccine and the fifth dose at age 5 years with at least 36 months between the fourth and fifth doses provided the best protection against pertussis among children and adolescents in this outbreak. PMID- 17133159 TI - Ciprofloxacin treatment in preterm neonates in Bangladesh: lack of effects on growth and development. AB - BACKGROUND: Quinolone-induced arthropathic toxicity in weight-bearing joints observed in juvenile animals during preclinical testing has largely restricted the routine use of ciprofloxacin in the pediatric age group. As histopathologic, radiologic and magnetic resonance imaging monitoring evidence has gathered supporting the safety of fluoroquinolones in children, many pediatricians have started to prescribe quinolones to some patients on a compassionate basis. OBJECTIVE: The objective of this study was to ascertain the safety of ciprofloxacin in preterm neonates <33 weeks gestational age treated at Dhaka Shishu (Children) Hospital in Bangladesh. METHODS: Long-term follow up was done to monitor the growth and development of preterm infants who were administered intravenous ciprofloxacin in the neonatal period. Ciprofloxacin was used only as a life-saving therapy in cases of sepsis produced by bacterial agents resistant to other antibiotics. Another group of preterm neonates with septicemia who were not exposed to ciprofloxacin, but effectively treated with other antibiotics and followed up, were matched with cases for gender, gestational age and birth weight and included as a comparison group. Forty-eight patients in the ciprofloxacin group and 66 patients in the comparison group were followed up for a mean of 24.7 +/- 18.5 months and 21.6 +/- 18.8 months, respectively. RESULTS: No osteoarticular problems or joint deformities were observed in the ciprofloxacin group during treatment or follow up. No differences in growth and development between the groups were found. CONCLUSIONS: Ciprofloxacin is a safe therapeutic option for newborns with sepsis produced by multiply resistant organisms. PMID- 17133160 TI - Salvage therapy with abacavir and other reverse transcriptase inhibitors for human immunodeficiency-associated encephalopathy. AB - BACKGROUND: HIV-associated encephalopathy (HIV-AE) is a severe neurologic condition that affects HIV-infected children. The potential benefit of antiretroviral (ARV) agents with good cerebrospinal fluid (CSF) penetration remains to be defined. Abacavir (ABC) achieves good CSF concentrations and studies of high-dose ABC showed benefit in adults with HIV dementia. The present study evaluated the safety and virologic, immunologic and neuropsychological responses of an ARV regimen including high-dose ABC in children with HIV-AE. METHODS: Children between 3 months and 18 years old and abacavir-naive with HIV AE and virologic failure were eligible. RESULTS: : Seventeen children (16 ARV experienced) were enrolled and 14 children completed 48 weeks of therapy. The overall tolerability was good; 2 children had a possible hypersensitivity reaction. At week 48, 53% and 59% of the children achieved HIV RNA levels below the limit of quantitation in plasma and CSF, respectively. The median (25%-75% range) change of HIV RNA from baseline to week 48 was -2.29 (-0.81 to -2.47) log10 copies/mL in plasma and -0.94 (0 to -1.13) log10 copies/mL in CSF. The mean increases in CD4 (+/-standard error of mean) cell count and CD4% were 427 (+/ 169) cells/mm and 8% (+/-2), respectively. Concentrations of soluble tumor necrosis factor receptor II were reduced in plasma and CSF. Children less than 6 years of age demonstrated significant neuropsychological improvement at week 48. CONCLUSIONS: In the present study with a limited number of children, highly active ARV therapy including high-dose ABC showed a safety profile similar to standard dose ABC and provided clinical, immunologic and virologic response in children with HIV-AE at week 48. Children less than 6 years of age also demonstrated significant neuropsychological improvement. PMID- 17133161 TI - Impact of rapid viral testing for influenza A and B viruses on management of febrile infants without signs of focal infection. AB - BACKGROUND: The objective of this study was to assess the effect of rapid testing for influenza virus on management of febrile young infants. METHODS: During 2 influenza seasons (November to December 2003 and December 2004 to February 2005), we studied prospectively infants who were 0 to 36 months of age who presented to the emergency department (ED) with fever in the absence of signs of focal infection. The Directigen Flu A+B test was used to determine infection with influenza virus types A or B. Confirmatory viral cultures were not done. RESULTS: Rapid influenza testing was performed in 206 infants and 84 (40.7%) of them were influenza-positive. Infants with a positive and a negative influenza test showed a similar mean (standard deviation) age (6.86 [6.3] versus 6.55 [6.8] months) and mean temperature (39.38 degrees C [0.6] versus 39.32 degrees C [0.8]), but there were significant differences (P < 0.01) in the percentage of patients undergoing blood tests (33.3% versus 100%), urinalysis (80.9% versus 100%), chest roentgenogram (14.2% versus 32%), cerebrospinal fluid analysis (1.33% versus 21.3%), mean length of stay in the ED (116.2 [75.5] versus 192.9 [76.3] minutes), admission to the ED observation ward (8.3% versus 21.3%), inpatient care (2.3% versus 16.4%) and antibiotic treatment (0% versus 38.5%). All positive bacterial cultures occurred among influenza-negative patients. CONCLUSIONS: The inclusion of rapid influenza testing for the evaluation of febrile young infants without signs of focal infection during influenza season decreases the need for additional studies and reduces the length of stay in the ED, the use of antibiotic treatment and unnecessary hospitalizations. PMID- 17133162 TI - Human papilloma virus in hyperplastic tonsillar and adenoid tissues in children. AB - BACKGROUND: Human papillomavirus (HPV) in childhood causes skin and anogenital warts as well as the recurrent respiratory papillomatosis, a life-threatening cause of upper airway obstruction in children. To date, the information on HPV infection in tonsillar and adenoid hyperplasia in children is limited. PURPOSE: The purpose of this study was to investigate the presence of HPV DNA in children with benign tonsillar and/or adenoid hyperplasia. METHODS: One hundred six samples of paraffin-embedded adenoid and/or tonsillar tissues from 102 children, 57 girls and 45 boys (age range, 2-14 years), were tested for the presence of HPV DNA using polymerase chain reaction (PCR) with general primers GP5+/GP6+. HPV typing was performed by PCR with specific primers for HPV-16, -18, -33 and -11. RESULTS: HPV DNA was detected in 9 (8.5%) of the 106 collected specimens. The frequencies of HPV typing were 6 of 9 (66.7%) for HPV-16, 2 of 9 (22.2%) for HPV 11, zero of 9 (0%) for HPV-33 and HPV-18, whereas one HPV-positive sample remained untyped. No multiple HPV infection was detected. HPV was detected in 6 (9.4%) children with tonsillar hyperplasia and in 3 (7.1%) with adenoid hyperplasia. The mean age of children with HPV-positive specimens was lower than that of HPV-negative children (P = 0.006). No statistical correlation in the prevalence of HPV infection was observed according to children's sex, origin or residence (urban or rural). CONCLUSION: Although the significance of the presence of HPV DNA in tonsillar and adenoid hyperplasias remains obscure, the PCR detection of high-risk HPV DNA should be evaluated cautiously. PMID- 17133163 TI - Breast milk reduces the risk of illness in children of mothers with cholera: observations from an epidemic of cholera in Guinea-Bissau. AB - BACKGROUND: A protective effect of breastfeeding against cholera has been demonstrated in areas endemic of cholera. To assess the protection offered by breast milk from mothers living in an area that had been free from cholera for 7 years, we investigated mothers with cholera and their children during an epidemic with Vibrio cholerae El Tor in the capital of Guinea-Bissau. METHODS: Eighty mothers with clinical cholera and their children were identified, and interviewed. Blood samples for vibriocidal and antitoxin antibodies were collected from mother-and-child pairs. Breast milk samples were collected from lactating mothers. Cholera was defined as acute watery diarrhea during the epidemic and a vibriocidal reciprocal titer of 20 or above. RESULTS: Three (7%) of 42 breastfed children had cholera as defined above compared with 9 (24%) of 38 nonbreastfed children (RR for breastfed children, 0.19; 95% CI, 0.04-0.91, adjusted for age). The 3 breastfed children who developed cholera received milk containing lower concentrations of anticholera toxin IgA/total IgA (median, 2.0 units/mL) than 14 children who had serologic signs of colonization but did not develop the disease (median, 17.4 units/mL). CONCLUSIONS: The protective effect of breast milk against cholera is not confined to endemic areas. Lactating mothers with cholera should receive supportive care to continue breastfeeding. PMID- 17133164 TI - The impact of a change in bacille Calmette-Guerin vaccine policy on tuberculosis incidence in children in Cape Town, South Africa. AB - BACKGROUND: A decision by the South African National Department of Health to change the route of administration and strain of bacille Calmette-Guerin (BCG) vaccine was implemented in Cape Town, South Africa, between July and December 2000. This provided an opportunity to compare the incidence of tuberculosis and proportion with disseminated disease in children less than 2 years old before and after the changeover from percutaneous (PC) Tokyo 172 BCG to intradermal (ID) 1331 Danish BCG immunization. METHODS: Clinical records of all tuberculosis patients aged less than 2 years at diagnosis and born between January 1, 1999, and June 30, 2000 (PC cohort) and between January 1, 2001, and June 30, 2002 (ID cohort) were collected. All cases were reviewed for likelihood of TB, its severity and disease dissemination. RESULTS: The number of reported patients with tuberculosis in the PC cohort was 1369 and in the ID cohort 1397, giving incidence rates of 866 (95% confidence interval [CI], 821-913) and 858 (95% CI, 814-904) per 100,000 person-years, respectively. The proportion who had disseminated disease (meningitis and/or miliary spread) was significantly lower in the ID cohort (4.7%) than in the PC cohort (8.6%) (relative risk, 0.54; 95% CI, 0.40-0.72). Those not vaccinated had a significantly higher proportion of disseminated disease cases (29.2%) than the PC and ID groups combined (6.6%) (relative risk, 4.4; 95% CI, 2.7-6.7). CONCLUSION: A program using Danish 1331 BCG given intradermally did not prevent more tuberculosis cases in children overall as compared with a program using Tokyo 172 BCG given percutaneously but reduced the proportion with disseminated disease. PMID- 17133165 TI - Prevention of malaria in children. PMID- 17133166 TI - Transient paresis associated with cat-scratch disease: case report and literature review of vertebral osteomyelitis caused by Bartonella henselae. AB - Cat-scratch disease (CSD) rarely presents as vertebral osteomyelitis. We describe a case with paresis of the arm with total recovery after antibiotic and neurosurgical therapy. We reviewed 20 other cases of CSD vertebral osteomyelitis in the literature. This diagnosis should be considered in patients with systemic symptoms, back pain, and cat contact. The prognosis is generally good. PMID- 17133167 TI - Vaccination usage among an old-order Amish community in Illinois. AB - The Old-Order Amish have low rates of vaccination and are at increased risk for vaccine-preventable diseases. A written survey was mailed to all Amish households in the largest Amish community in Illinois inquiring about their vaccination status and that of their children. In this survey, the Amish do not universally reject vaccines, adequate vaccination coverage in Amish communities can be achieved, and Amish objections to vaccines might not be for religious reasons. PMID- 17133168 TI - Comparison of trip characteristics of children and adults with travel-acquired hepatitis A infection. AB - We compared the trip characteristics of 84 child and 99 adult cases with travel acquired hepatitis A (HA). Most pediatric cases had traveled in Asia for more than 30 days and had stayed and eaten most of their meals in the homes of friends and relatives in a country where they had not been born. In contrast, the adults with travel-acquired HA had visited Latin America or the Caribbean for 14 days or less and had stayed primarily in hotels. Specific public health interventions should be undertaken to prevent HA in traveling children. PMID- 17133169 TI - Efficacy and safety of caspofungin therapy in children with invasive fungal infections. AB - Twenty children with proven (n = 12) or probable (n = 8) invasive fungal infections received caspofungin treatment either as first-line (n = 7) or as salvage (n = 13) therapy and as monotherapy (n = 5) or in combination (n = 15). Eleven had aspergillosis, 7 had candidiasis, and 2 had Rhodotorula infections. Caspofungin was well tolerated. Nine patients experienced 11 drug-related adverse events, none were severe, and none led to drug discontinuation. Caspofungin as a first-line treatment was successful in 5 of the 7 children (these 5 patients survived the infectious episode, with a follow-up of 147 days), and salvage therapy rescued 8 of 13 children, but only 5 of them survived. PMID- 17133170 TI - The risk of respiratory syncytial virus-related hospitalizations in preterm infants of 29 to 35 weeks' gestational age. AB - Among 1158 preterm infants of 29-35 weeks' gestational age, respiratory syncytial virus (RSV) -related hospitalizations (RSV-H) occurred in 4.2% during the first year of life. Four independent factors influenced the risk for RSV-H: neurologic problems (odds ratio [OR], 3.6), male gender (OR, 2.8), presence of an older sibling (OR, 1.7) and discharge from October to December (OR, 1.7). The estimated risk of RSV-H varied between 1% (no risk factor present) and 30% (4 risk factors present). PMID- 17133171 TI - Lobar pneumonia caused by nontyphoidal Salmonella in a Malawian child. AB - Nontyphoidal Salmonella (NTS) is recognized as a common cause of bacteremia in malaria-endemic Africa but its importance as a cause of pneumonia is uncertain. We report a case of pneumonia caused by NTS confirmed by culture of lung aspirate from a consolidated left lung in a 16 month-old HIV-uninfected girl who had been admitted to the hospital 1 month previously with severe malaria. She did not respond to first-line antibiotic therapy for benzylpenicillin and gentamicin but improved with ceftriaxone therapy. PMID- 17133172 TI - Human immunodeficiency virus infection and amebiasis. AB - Pediatric human immunodeficiency virus (HIV) infection is a major and increasing burden worldwide, but particularly in sub-Saharan Africa. Coinfection with other pathogens increases the likelihood of progression of HIV/acquired immunodeficiency syndrome (AIDS), and the immunosuppressive consequences of the disease predispose to opportunistic infections that can run a fulminant course. Despite high prevalence, amebiasis has not appeared as a major source of morbidity during the HIV/AIDS pandemic. Information from recent sources, however, appears to suggest that amebiasis may indeed be a risk for individuals living with HIV/AIDS. PMID- 17133173 TI - Detection of human influenza virus in the stool of children. PMID- 17133174 TI - Mean peroxidase index as a potential marker for neonatal infection. PMID- 17133175 TI - Streptococcus pneumoniae clinical isolate highly resistant to fluoroquinolones in a child. PMID- 17133176 TI - Apolipoprotein E4 polymorphism as a genetic predisposition to delirium in critically ill patients. AB - OBJECTIVE: To test for an association between apolipoprotein E (APOE) genotypes and duration of intensive care unit delirium. DESIGN: Prospective, observational cohort study. SETTING: A 541-bed, community-based teaching hospital. PATIENTS: Fifty-three mechanically ventilated intensive care unit patients. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: All patients were managed with standardized sedation and ventilator weaning protocols as part of an ongoing clinical trial and were evaluated prospectively for delirium with the Confusion Assessment Method for the Intensive Care Unit (CAM-ICU). DNA was extracted from whole blood samples obtained on enrollment, and APOE genotype was determined using polymerase chain reaction followed by restriction enzyme digestion by investigators blinded to the clinical information. Delirium occurred in 47 (89%) patients at some point during the intensive care unit stay. Of the 53 patients, 12 (23%) had an APOE4 allele (APOE4+) and 41 (77%) had only APOE2 or APOE3 alleles (APOE4-). APOE4+ patients were younger (53.2 +/- 21.9 vs. 65.4 +/- 13.4, p = .08) and less often admitted for pneumonia (0% vs. 29.3%, p = .05) compared with APOE4- patients, yet they had a duration of delirium that was twice as long: median (interquartile range), 4 (3, 4.5) vs. 2 (1, 4) days (p = .05). No other clinical outcomes were significantly different between the APOE4+ and APOE4- patients. Using multivariable regression analysis to adjust for age, admission diagnosis of sepsis or acute respiratory distress syndrome or pneumonia, severity of illness, and duration of coma, the presence of APOE4 allele was the strongest predictor of delirium duration (odds ratio, 7.32; 95% confidence interval, 1.82-29.51, p = .005). CONCLUSIONS: APOE4 allele represents the first demonstrated genetic predisposition to longer duration of delirium in humans. PMID- 17133177 TI - A multiple-center survey on the use in clinical practice of noninvasive ventilation as a first-line intervention for acute respiratory distress syndrome. AB - OBJECTIVE: In randomized studies of heterogeneous patients with hypoxemic acute respiratory failure, noninvasive positive pressure ventilation (NPPV) was associated with a significant reduction in endotracheal intubation. The role of NPPV in patients with acute respiratory distress syndrome (ARDS) is still unclear. The objective was to investigate the application of NPPV as a first-line intervention in patients with early ARDS, describing what happens in everyday clinical practice in centers having expertise with NPPV. DESIGN: Prospective, multiple-center cohort study. SETTING: Three European intensive care units having expertise with NPPV. PATIENTS: Between March 2002 and April 2004, 479 patients with ARDS were admitted to the intensive care units. Three hundred and thirty-two ARDS patients were already intubated, so 147 were eligible for the study. INTERVENTIONS: Application of NPPV. MEASUREMENTS AND MAIN RESULTS: NPPV improved gas exchange and avoided intubation in 79 patients (54%). Avoidance of intubation was associated with less ventilator-associated pneumonia (2% vs. 20%; p < .001) and a lower intensive care unit mortality rate (6% vs. 53%; p < .001). Intubation was more likely in patients who were older (p = .02), had a higher Simplified Acute Physiology Score (SAPS) II (p < .001), or needed a higher level of positive end-expiratory pressure (p = .03) and pressure support ventilation (p = .02). Only SAPS II >34 and a Pao2/Fio2 < or =175 after 1 hr of NPPV were independently associated with NPPV failure and need for endotracheal intubation. CONCLUSIONS: In expert centers, NPPV applied as first-line intervention in ARDS avoided intubation in 54% of treated patients. A SAPS II >34 and the inability to improve Pao2/Fio2 after 1 hr of NPPV were predictors of failure. PMID- 17133178 TI - Advanced age exacerbates the pulmonary inflammatory response after lipopolysaccharide exposure. AB - OBJECTIVE: The aged population is at a higher risk of mortality as a result of complications of injury or infection, such as acute lung injury. The objective of this study was to analyze pulmonary inflammatory responses in young and aged mice after administration of lipopolysaccharide. DESIGN: Prospective, controlled laboratory study. SETTING: Animal resource facilities and research laboratory. SUBJECTS: Young (2-3 months old) and aged (18-20 months old) female BALB/c mice. INTERVENTIONS: Animals received intraperitoneal injection of lipopolysaccharide derived from Pseudomonas aeruginosa. Control mice received saline alone. After 24 hrs, mice were killed. Pulmonary neutrophil infiltration was assessed histologically and by myeloperoxidase activity. Pulmonary levels of the CXC chemokines, monocyte inflammatory protein-2 and KC, and cytokines, tumor necrosis factor-alpha and interleukin-1beta, were assessed by enzyme-linked immunosorbent assay. MEASUREMENTS AND MAIN RESULTS: Lungs of aged mice given lipopolysaccharide showed a six-fold higher neutrophil infiltration and three-fold higher level of myeloperoxidase activity than lungs of young mice given lipopolysaccharide. Pulmonary levels of monocyte inflammatory protein-2 and KC were significantly higher in the lungs of aged mice given lipopolysaccharide, compared with younger mice. Levels of tumor necrosis factor-alpha and interleukin-1beta in the lung were analyzed as well. After lipopolysaccharide treatment, there was no difference in the level of tumor necrosis factor-alpha in lungs of young and aged animals, but interleukin-1beta was two-fold higher in the lungs of the aged group. These data suggest that at this time point, interleukin-1beta may contribute to the higher production of CXC chemokines observed in lungs of aged mice vs. young mice receiving lipopolysaccharide. CONCLUSIONS: The hyperreactive systemic inflammatory response seen in aged individuals after lipopolysaccharide administration is accompanied by an exacerbated pulmonary inflammatory response, which may contribute to the higher mortality seen in the aged given an inflammatory insult. PMID- 17133179 TI - Protective effects of atorvastatin in rat models of acute pulmonary embolism: involvement of matrix metalloproteinase-9. AB - OBJECTIVE: Matrix metalloproteinases (MMPs) have been implicated in the pathophysiology of acute pulmonary embolism (APE)-induced pulmonary hypertension. Here, we evaluate the effects of atorvastatin pretreatment on APE-induced pulmonary hypertension, 24-hr mortality rate, and changes in plasma and lung MMP 2 and MMP-9 activities. DESIGN: Controlled animal study. SETTING: University research laboratory. SUBJECTS: Male Wistar rats. INTERVENTIONS: Rats received atorvastatin (30 mg/kg/day orally) or tap water for 2 wks. In study 1, we examined whether atorvastatin affected APE-induced pulmonary hypertension by using a rat isolated lung perfusion model of APE. In study 2, we examined whether atorvastatin affects the survival rate after APE, which was induced by rapid intravenous injection of 14 mg/kg of a suspension of microspheres (or saline) into the tail vein. MEASUREMENTS AND MAIN RESULTS: Plasma nitrite/nitrate concentrations were measured by chemiluminescence. Pretreatment with atorvastatin was associated with 49% higher nitrite/nitrate levels compared with controls (p < .05). In study 1, whereas APE increased mean pulmonary artery pressure (MPAP) by 13.0 +/- 1.6 mm Hg in perfused lungs isolated from rats pretreated with water, pretreatment with atorvastatin attenuated by 27% the increases in MPAP after APE. In study 2, pretreatment with atorvastatin was associated with a significant increase in 24-hr survival rate after APE, which was 48% in embolized rats pretreated with water and 64% in rats pretreated with atorvastatin (p < .05). Gelatin zymography of lung and plasma MMP-2 and MMP-9 was performed. Lungs and plasma from embolized rats showed higher levels of both pro- and activated forms of MMP-9 compared with those from nonembolized animals (all p < .05). However, pretreatment with atorvastatin attenuated by 32% the increases in lung-activated MMP-9 levels after APE (p < .05). CONCLUSIONS: These results suggest that pretreatment with atorvastatin attenuates APE-induced pulmonary hypertension and increases 24-hr survival rate by mechanisms that result in attenuated increases in lung activated MMP-9 after APE. PMID- 17133180 TI - Tracheostomy does not improve the outcome of patients requiring prolonged mechanical ventilation: a propensity analysis. AB - OBJECTIVE: To examine the association between the performance of a tracheostomy and intensive care unit and postintensive care unit mortality, controlling for treatment selection bias and confounding variables. DESIGN: Prospective, observational, cohort study. SETTING: Twelve French medical or surgical intensive care units. PATIENTS: Unselected patients requiring mechanical ventilation for > or =48 hrs enrolled between 1997 and 2004. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Two models of propensity scores for tracheostomy were built using multivariate logistic regression. After matching on these propensity scores, the association of tracheostomy with outcomes was assessed using multivariate conditional logistic regression. Results obtained with the two models were compared. Of the 2,186 patients included, 177 (8.1%) received a tracheostomy. Both models led to similar results. Tracheostomy did not improve intensive care unit survival (model 1: odds ratio, 0.94; 95% confidence interval, 0.63-1.39; p = .74; model 2: odds ratio, 1.12; 95% confidence interval, 0.75-1.67; p = .59). There was no difference whether tracheostomy was performed early (within 7 days of ventilation) or late (after 7 days of ventilation). In patients discharged free from mechanical ventilation, tracheostomy was associated with increased postintensive care unit mortality when the tracheostomy tube was left in place (model 1: odds ratio, 3.73; 95% confidence interval, 1.41-9.83; p = .008; model 2: odds ratio, 4.63; 95% confidence interval, 1.68-12.72, p = .003). CONCLUSIONS: Tracheostomy does not seem to reduce intensive care unit mortality when performed in unselected patients but may represent a burden after intensive care unit discharge. PMID- 17133181 TI - Influence of drotrecogin alpha (activated) infusion on the variation of Bax/Bcl-2 and Bax/Bcl-xl ratios in circulating mononuclear cells: a cohort study in septic shock patients. AB - OBJECTIVE: Drotrecogin alpha (activated) (DAA), or recombinant human activated protein C, is a new treatment in sepsis-induced multiple organ failure, leading to significant reduction in the mortality rate, thanks to its anticoagulant properties. It has been suggested that DAA has anti-inflammatory and antiapoptotic effects in sepsis animal models. This study investigates the potential actions of DAA on circulating mononuclear cells apoptosis in human septic shock. DESIGN: Prospective, cohort study. SETTING: Two intensive care wards and two research laboratories in a university hospital. PATIENTS: Twenty two septic shock patients with DAA treatment (DAA+), 19 septic shock patients without DAA treatment (DAA-), and 14 healthy controls were successively enrolled, but only 20 DAA+ and 16 DAA- patients fulfilled criteria for statistical analysis. INTERVENTIONS: Blood samples were collected at inclusion and 24 hrs later. MEASUREMENTS AND MAIN RESULTS: Circulating mononuclear cell apoptosis levels were assessed by flow cytometry with annexin V, and variations of the apoptotic rheostats (Bax/Bcl-2 and Bax/Bcl-xl ratios) were analyzed by real-time reverse transcription-polymerase chain reaction. Apoptosis was significantly increased in septic shock patients (DAA+, 12 +/- 6.4%; DAA-, 10.4 +/- 5%) vs. healthy patients (3.4 +/- 2.1%, p < .001). Twenty-four hours after DAA infusion, apoptosis was significantly lower in the DAA+ group compared with DAA- ones (respectively, 11.7 +/- 5.3% and 16.2 +/- 7.6%, p < .001). At inclusion, DAA+ and DAA- groups showed comparable Bax/Bcl-2 ratio (DAA+, 0.92 +/- 0.9; DAA-, 1.32 +/- 0.87) and Bax/Bcl-xl ratio (DAA+, 2 +/- 1.04; DAA-, 1.31 +/- 0.93). In contrast, 24 hrs later we observed a significant decrease in these ratios, indicating an antiapoptotic effect in the DAA+ group (Bax/Bcl-2, 0.39 +/- 0.27; Bax/Bcl-xl, 0.68 +/- 0.35) compared with the DAA- group (Bax/Bcl-2, 1.81 +/- 1.1; Bax/Bcl-xl, 1.22 +/- 0.92, p = .001 and p = .039, respectively). CONCLUSIONS: In vivo, in human septic shock, DAA has antiapoptotic effects on circulating mononuclear cells, assessed by a significant decrease of both the Bax/Bcl-2 and Bax/Bcl-xl ratios. PMID- 17133182 TI - Polymorphisms in the mannose binding lectin-2 gene and acute respiratory distress syndrome. AB - OBJECTIVE: The variant alleles in the mannose binding lectin-2 (MBL-2) gene have been associated with MBL deficiency and increased susceptibility to sepsis. We postulate that the variant MBL-2 genotypes are associated with increased susceptibility to and mortality in acute respiratory distress syndrome (ARDS). DESIGN: Nested case-control study. SETTING: Tertiary academic medical center. PATIENTS: Two hundred and twelve Caucasians with ARDS and 442 controls genotyped for the variant X, D, B, and C alleles of codon -221, 52, 54, and 57, respectively. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients homozygous for the variant codon 54B allele (54BB) had worse severity of illness on admission (p = .007), greater likelihood of septic shock (p = .04), and increased odds of ARDS (adjusted odds ratio, 6.7; 95% confidence interval, 1.5 31) when compared with heterozygotes and homozygotes for the wild-type allele. This association with ARDS was especially strong among the 311 patients with septic shock (adjusted odds ratio, 12.0; 95% confidence interval, 1.9-74). Among the patients with ARDS, the 54BB genotype was associated with more daily organ dysfunction (p = .01) and higher mortality (adjusted hazard rate, 4.0; 95% confidence interval, 1.6-10). Development of ARDS and outcomes in ARDS did not vary significantly with variant alleles of codon -221, 52, and 57, but the power to detect an effect was limited secondary to the low allele frequencies. CONCLUSIONS: The MBL-2 codon 54BB genotype may be important in ARDS susceptibility and outcome. Additional studies are needed to confirm these findings in other populations. PMID- 17133183 TI - Early activity is feasible and safe in respiratory failure patients. AB - OBJECTIVE: To determine whether early activity is feasible and safe in respiratory failure patients. DESIGN: Prospective cohort study. SETTING: From June 1, 2003, through December 31, 2003, we assessed safety and feasibility of early activity in all consecutive respiratory failure patients who required mechanical ventilation for >4 days admitted to our respiratory intensive care unit (RICU). A majority of patients were treated in another intensive care unit (ICU) before RICU admission. We excluded patients who required mechanical ventilation for < or =4 days. PATIENTS: Eight-bed RICU at LDS Hospital. INTERVENTIONS: We assessed patients for early activity as part of routine respiratory ICU care. We prospectively recorded activity events and adverse events. We defined three activity events as sit on bed, sit in chair, and ambulate. We defined six activity-related adverse events as fall to knees, tube removal, systolic blood pressure >200 mm Hg, systolic blood pressure <90 mm Hg, oxygen desaturation <80%, and extubation. MEASUREMENTS AND MAIN RESULTS: During the study period, we conducted a total of 1,449 activity events in 103 patients. The activity events included 233 (16%) sit on bed, 454 (31%) sit in chair, and 762 (53%) ambulate. In patients with an endotracheal tube in place, there were a total of 593 activity events, of which 249 (42%) were ambulation. There were <1% activity-related adverse events, including fall to the knees without injury, feeding tube removal, systolic blood pressure >200 mm Hg, systolic blood pressure <90 mm Hg, and desaturation <80%. No patient was extubated during activity. CONCLUSIONS: We conclude that early activity is feasible and safe in respiratory failure patients. A majority of survivors (69%) were able to ambulate >100 feet at RICU discharge. Early activity is a candidate therapy to prevent or treat the neuromuscular complications of critical illness. PMID- 17133184 TI - Severe stunned myocardium after lightning strike. AB - OBJECTIVE: To report the development of myocardial stunning and severe heart failure after lightning strike with total recovery of function. DESIGN: Case report. SETTING: Coronary care unit at Medica Sur Clinic, Mexico. PATIENT: A 42 yr-old woman who was hit by lightning developed rapid and progressive hemodynamic deterioration manifested by cardiogenic shock that required invasive monitoring. Twenty-four hours after the strike, intravenous levosimendan and intra-aortic balloon pump were initiated because the patient demonstrated no significant response to management with conventional inotropic agents. Two days later, echocardiographic signs of systolic and diastolic dysfunction improved markedly. Dual-isotope-imaging myocardial perfusion testing with technetium-99m-sestamibi and thallium-201, performed 9 days after admission, showed normal perfusion and normal left ventricular systolic function. The patient exhibited complete recovery of function. The exact mechanism of abnormal contractility in the absence of direct electrofulguration is unknown but may be explained by release of oxygen free radicals, proteolysis of the contractile apparatus, and cytosolic overload of intracellular calcium, followed by reduced myofilament sensitivity to calcium. These abnormalities are consistent with stunned myocardium. CONCLUSIONS: Lightning strike may cause serious contractile dysfunction in the absence of irreversible myocardial injury, but the exact mechanism of this phenomenon remains unknown. We propose that lighting strike can cause myocardial stunning resulting in severe but reversible left ventricular dysfunction. The patient's recovery was facilitated by support treatment including administration of levosimendan, which increases the intracellular sensitivity to calcium, a mechanism disturbed in patients with myocardial stunning. PMID- 17133185 TI - High-frequency oscillatory ventilation following prone positioning prevents a further impairment in oxygenation. AB - OBJECTIVE: The improvement in oxygenation with prone positioning is not persistent when patients with acute respiratory distress syndrome (ARDS) are turned supine. High-frequency oscillatory ventilation (HFOV) aims to maintain an open lung volume by the application of a constant mean airway pressure. The aim of this study was to show that HFOV is able to prevent the impairment in oxygenation when ARDS patients are turned back from the prone to the supine position. DESIGN: Prospective, comparative randomized study. SETTING: A medical intensive care unit. PATIENTS: Forty-three ARDS patients with a Pao2/Fio2 ratio <150 at positive end-expiratory pressure > or =5 cm H2O. INTERVENTIONS: After an optimization period, the patients were assigned to one of three groups: a) conventional lung-protective mechanical ventilation in the prone position (12 hrs) followed by a 12-hr period of conventional lung-protective mechanical ventilation in the supine position (CV(prone)-CV(supine)); b) conventional lung protective mechanical ventilation in the supine position (12 hrs) followed by HFOV in the supine position (12 hrs) (CV(supine)-HFOV(supine)); or c) conventional lung-protective mechanical ventilation in the prone position (12 hrs) followed by HFOV in the supine position (CV(prone)-HFOV(supine) group). MEASUREMENTS AND MAIN RESULTS: Pao2/Fio2 ratio was higher at the end of the study period in the CV(prone)-HFOV(supine) group than in the CV(prone)-CV(supine) group (p < .02). Venous admixture at the end of the study period was lower in the CV(prone)-HFOV(supine) group than in the two other groups. CONCLUSIONS: HFOV maintained the improvement in oxygenation related to prone positioning when ARDS patients were returned to the supine position. PMID- 17133186 TI - Vasopressin: mechanisms of action on the vasculature in health and in septic shock. AB - BACKGROUND: Vasopressin is essential for cardiovascular homeostasis, acting via the kidney to regulate water resorption, on the vasculature to regulate smooth muscle tone, and as a central neurotransmitter, modulating brainstem autonomic function. Although it is released in response to stress or shock states, a relative deficiency of vasopressin has been found in prolonged vasodilatory shock, such as is seen in severe sepsis. In this circumstance, exogenous vasopressin has marked vasopressor effects, even at doses that would not affect blood pressure in healthy individuals. These two findings provide the rationale for the use of vasopressin in the treatment of septic shock. However, despite considerable research attention, the mechanisms for vasopressin deficiency and hypersensitivity in vasodilatory shock remain unclear. OBJECTIVE: To summarize vasopressin's synthesis, physiologic roles, and regulation and then review the literature describing its vascular receptors and downstream signaling pathways. A discussion of potential mechanisms underlying vasopressin hypersensitivity in septic shock follows, with reference to relevant clinical, in vivo, and in vitro experimental evidence. DATA SOURCE: Search of the PubMed database (keywords: vasopressin and receptors and/or sepsis or septic shock) for articles published in English before May 2006 and manual review of article bibliographies. DATA SYNTHESIS AND CONCLUSIONS: The pathophysiologic mechanism underlying vasopressin hypersensitivity in septic shock is probably multifactorial. It is doubtful that this phenomenon is merely the consequence of replacing a deficiency. Changes in vascular receptors or their signaling and/or interactions between vasopressin, nitric oxide, and adenosine triphosphate-dependent potassium channels are likely to be relevant. Further translational research is required to improve our understanding and direct appropriate educated clinical use of vasopressin. PMID- 17133187 TI - Open and closed endotracheal suction systems in mechanically ventilated intensive care patients: a meta-analysis. AB - BACKGROUND: Closed suction systems (CSS) are increasingly replacing open suction systems (OSS) to perform endotracheal toilet in mechanically ventilated intensive care unit patients. Yet effectiveness regarding patient safety and costs of these systems has not been carefully analyzed. OBJECTIVE: To review effectiveness of CSS and OSS, with respect to patient outcome, bacterial contamination, and costs in adult intensive care unit patients. DATA SOURCE: Search of MEDLINE, CINAHL, EMBASE, and Cochrane databases and a manual review of article bibliographies. STUDY SELECTION: Randomized controlled trials comparing CSS and OSS in adult intensive care unit patients were retrieved. DATA EXTRACTION/SYNTHESIS: Assessment of abstracts and study quality was performed by two reviewers. Data were combined in meta-analyses by random effect models. Fifteen trials were identified. No significant differences were found in incidences of ventilator associated pneumonia (eight studies, 1,272 patients) and mortality (four studies, 1,062 patients). No conclusions could be drawn with respect to arterial oxygen saturation (five studies, 109 patients), arterial oxygen tension (two studies, 19 patients), and secretion removal (two studies, 37 patients). Compared with OSS, endotracheal suctioning with CSS significantly reduced changes in heart rate (four studies, 85 patients; weighted mean difference, -6.33; 95% confidence interval, -10.80 to -1.87) and changes in mean arterial pressure (three studies, 59 patients; standardized mean difference, -0.43; 95% confidence interval, -0.87 to 0.00) but increased colonization (two studies, 126 patients; relative risk, 1.51; 95% confidence interval, 1.12-2.04). CSS seems to be more expensive than OSS. CONCLUSIONS: Based on the results of this meta-analysis, there is no evidence to prefer CSS more than OSS. PMID- 17133188 TI - Mechanisms of a reduced cardiac output and the effects of milrinone and levosimendan in a model of infant cardiopulmonary bypass. AB - OBJECTIVES: A low cardiac output state is an important cause of morbidity after pediatric cardiopulmonary bypass. The objectives of our study were to define the early precipitants of the reduced cardiac output and to investigate the effects on these of milrinone and levosimendan in a model of pediatric cardiopulmonary bypass. DESIGN: Experimental study. SETTING: : Research laboratory at a university-affiliated, tertiary pediatric center. SUBJECTS: Eighteen piglets. INTERVENTIONS: Piglets, instrumented with systemic, pulmonary arterial, and coronary sinus catheters, pulmonary and circumflex arterial flow probes, and a left ventricular conductance-micromanometer-tipped catheter, underwent cardiopulmonary bypass with aortic cross-clamp and cardioplegic arrest. At 120 mins, they were assigned to control, milrinone, or levosimendan groups and studied for a further 120 mins. MEASUREMENTS AND MAIN RESULTS: In controls, between 120 and 240 mins, cardiac output decreased by 15%. Systemic vascular resistance was unchanged, but pulmonary vascular resistance increased by 19%. Systemic arterial elastance increased by 17%, indicating increased afterload. End systolic elastance was unchanged, and coronary sinus oxygen tension decreased by 4.0 +/- 1.7 mm Hg. In animals receiving milrinone cardiac output was preserved, and in animals receiving levosimendan cardiac output increased by 14%. Both drugs prevented an increase in arterial elastance and pulmonary vascular resistance after cardiopulmonary bypass. Systemic vascular resistance decreased by 31% after levosimendan, and end-systolic elastance increased by 48%, indicating improved contractility. Both agents prevented a decrease in coronary sinus oxygen tension. CONCLUSIONS: Increased afterload, which is not matched by an equivalent elevation in contractility, contributes to the reduced cardiac output early after pediatric cardiopulmonary bypass in this model. This increase is prevented by milrinone and levosimendan. Both agents exert additional beneficial effects on pulmonary vascular resistance and myocardial oxygen balance, although levosimendan has greater inotropic properties. PMID- 17133189 TI - Refinement, scoring, and validation of the Family Satisfaction in the Intensive Care Unit (FS-ICU) survey. AB - OBJECTIVES: To refine the Family Satisfaction in the Intensive Care Unit (FS-ICU) survey and develop a validated method for scoring the instrument. DESIGN: Instrument development study, using data from two prospective cohort studies. SETTING: Intensive care units in seven university-affiliated hospitals (six Canadian, one United States). SUBJECTS: Family members of ICU patients. INTERVENTIONS: Based on a priori criteria, items were tagged for potential removal and discussed with the FS-ICU developers. Factor analysis was used to test the conceptual structure of the instrument and develop a scoring method based on scales and subscales. The new scoring method was validated in the U.S. cohort using the Quality of Dying and Death (QODD) instrument and nurse-assessed quality indicators. MEASUREMENTS AND MAIN RESULTS: A total of 1,038 family members completed the FS-ICU across seven sites. Fifteen items were initially tagged for possible removal. After consensus with the developers, ten items were dropped (and 24 were retained in the final instrument). Factor analysis explained 61.3% of the total variance using a two-factor model. The first factor pertained to satisfaction with care (14 items). The second factor encompassed satisfaction with decision making (10 items). A scoring method was developed based on this conceptual model. In validity testing, the FS-ICU was significantly correlated with the Family-QODD total score (Spearman's .56, p < .001) as well as individual QODD items such as quality of care by all providers (.64, p < .001). The FS-ICU also correlated significantly with multiple nurse-assessed quality indicators. CONCLUSIONS: The shortened FS-ICU measures two main conceptual domains satisfaction with care and satisfaction with decision making. Scores on the FS ICU show good validity against other indicators of ICU quality. The instrument holds promise as a useful outcome measure in studies that attempt to improve this component of ICU care. PMID- 17133190 TI - Enhancing Adherence: Proceedings of the state of science meeting on intervention research to improve ARV adherence, November 11-12, 2005, New Haven, Connecticut, USA. PMID- 17133191 TI - Involving behavioral scientists, health care providers, and HIV-infected patients as collaborators in theory-based HIV prevention and antiretroviral adherence interventions. AB - Health care providers are often hesitant to attempt health behavior change interventions with patients, although such interventions are frequently needed. When provider-initiated health behavior change interventions are attempted, they are often based on intuition or consist solely of delivering information and are insufficient to change behavior, rather than being based on well-validated and effective behavior change models. We argue that provider-initiated health behavior change interventions are effective and efficient if they are based on appropriate empirically validated theoretical models and developed in collaboration with behavioral scientists and patients. We present a new model for developing such collaborative interventions and initial evidence for its success. PMID- 17133192 TI - A practical method to calibrate self-reported adherence to antiretroviral therapy. AB - OBJECTIVE: Self-report of antiretroviral medications adherence is inexpensive and simple to use in clinical settings but grossly overestimates adherence. We investigated methods to calibrate patients' self-reported adherence to match objectively measured adherence more closely for the purpose of developing a practical and more accurate self-reported adherence measure. DESIGN: Longitudinal cohort design. METHODS: Using data from 2 prospective longitudinal clinical investigations conducted at 5 HIV clinics, we examined the discrepancy between self-reported adherence and objectively measured adherence. We evaluated the relation between attitudinal measures and the degree of discrepancy and used a cross-validation approach to propose candidate items to improve adherence survey methodology. RESULTS: Among 330 patients, self-reported adherence was consistently higher than objectively measured adherence. The best calibration models included the patient's self-reported adherence, duration of the antiretroviral regimen, and attitudinal measures (ability to take medication as instructed, believing medication can help one to live longer, whether or not it is too troublesome to take antiretrovirals, and feeling things are going the right way). CONCLUSION: The method efficiently identified survey items to improve self-reported adherence measurement. The calibrated measure more closely approximates objectively measured adherence and is more sensitive for detecting nonadherence. These models merit evaluation in other settings. PMID- 17133193 TI - Cost-effectiveness of an intervention to improve adherence to antiretroviral therapy in HIV-infected patients. AB - Adherence to antiretroviral medications has been shown to be an important factor in predicting viral suppression and clinical outcomes. The objective of this analysis was to assess the cost-effectiveness of a nursing intervention on antiretroviral adherence using data from a randomized controlled clinical trial as input to a computer-based simulation model of HIV disease. For a cohort of HIV infected patients similar to those in the clinical trial (mean initial CD4 count of 319 cells/mm), implementing the nursing intervention in addition to standard care yielded a 63% increase in virologic suppression at 48 weeks. This produced increases in expected survival (from 94.5 to 100.9 quality-adjusted life months) and estimated discounted direct lifetime medical costs ($253,800 to $261,300). The incremental cost-effectiveness ratio for the intervention was $14,100 per quality-adjusted life year gained compared with standard care. Adherence interventions with modest effectiveness are likely to provide long-term survival benefit to patients and to be cost-effective compared with other uses of HIV care funds. PMID- 17133194 TI - Adherence to antiretroviral medication among HIV-positive patients in Thailand. AB - BACKGROUND: The use of antiretroviral (ARV) medications is expanding rapidly in Thailand. The determinants of optimal adherence for HIV-positive patients in Thailand are unknown. METHODS: A sample of 149 Thai patients receiving ARV therapy at Bhumrasnaradura Infectious Disease Institute located near Bangkok completed a structured questionnaire and reported medication adherence on a 30 day visual analog scale. HIV RNA test results were abstracted from the medical record. RESULTS: Adherence ranged from 25% to 100%. The median was 100% and the mean was 96%. The majority of subjects (114, 77%) had an HIV RNA /=95% (odds ratio [OR] = 3.0; 95% confidence interval [CI] 1.3 to 7.1; P = 0.02) and with a lower mean number of months on ARV therapy (22 versus 32 months; P = 0.03). Gender, educational level, method of payment, use of GPO-VIR, and whether or not the patient was on his or her initial ARV regimen were not associated with an undetectable viral load. In the multivariate analysis, only length of time in months was associated with an undetectable viral load. For each additional month, the odds of being undetectable were 0.975. (OR = 0.975; 95% CI 0.954 to .996; P = 0.02). CONCLUSIONS: Adherence was high in this cohort and was associated with HIV RNA levels. However, these data confirm that adherence is only 1 factor that determines the effectiveness of ARV treatment. Duration of treatment was associated with virologic failure, controlling for adherence. PMID- 17133195 TI - Antiretroviral therapy in resource-poor settings. Decreasing barriers to access and promoting adherence. AB - Since 2002, the HIV Equity Initiative of the nongovernmental organization Partners in Health has been expanded in conjunction with the Haitian MOH to cover 7 public clinics. More than 8000 HIV-positive persons, 2300 of whom are on antiretroviral therapy (ART) are now followed. This article describes the interventions to promote access to care and adherence to ART developed in reference to the specific context of poverty in rural Haiti. User fees for clinic attendance have been waived for all patients with HIV and tuberculosis and for women presenting for prenatal services. Additionally, HIV testing has been integrated into the provision of primary care services to increase HIV case finding among those presenting to clinic because of illness, rather than solely focusing on those who present for voluntary counseling and testing (VCT). Once a patient is diagnosed with HIV, medications and monitoring tests are provided free of charge and transportation costs for follow-up appointments are covered to defray patients' out-of-pocket expenses. Patients are given home-based adherence support from a network of health workers who provide psychosocial support and directly observed therapy. In addition, the neediest patients receive nutritional support. Following the description of the program is an approximation of the costs of these interventions and a discussion of their impact. PMID- 17133196 TI - Treatment supporter to improve adherence to antiretroviral therapy in HIV infected South African adults. A qualitative study. AB - Little is known about the patient characteristics, social support networks, and relationship factors associated with excellent adherence in resource-limited settings, even though these can be important clues that inform the identification of targets and the approaches of individual- and community-based antiretroviral therapy adherence interventions. In this study, we aimed to understand how patient-selected treatment supports might affect antiretroviral treatment outcomes and to identify key components of support, including the social and material resources necessary for promoting high adherence in South Africa. Data were collected from the proceedings of 2 focus groups with 6 HIV-infected adults each and from 7 in-depth interviews with health care providers in the Western Cape Province of South Africa. The patients and health care workers identified individuals-usually a mother, daughter, sister, brother, or partner-who were confidantes and had moral authority with them. These individuals command respect, and patients allow them to influence health-related decision making, both of which are necessary if they are to be effective treatment supporters. Barriers to adherence identified by study participants include alcohol abuse, stigma related to disclosure of HIV status, and lack of financial resources and food. These are critically important challenges to address if high adherence is to be achieved in this setting. In addition, our results suggest that interventions tailored to treatment supporter characteristics and relationship factors may be effective in influencing patients' antiretroviral therapy adherence. PMID- 17133197 TI - Modified directly observed therapy to facilitate highly active antiretroviral therapy adherence in Beira, Mozambique. Development and implementation. AB - As resource-limited countries expand access to highly active antiretroviral therapy (HAART) treatment, innovative programs are needed to support adherence in the context of significant health system barriers. Modified directly observed therapy (mDOT) is one such strategy, but little is known about the process of designing and implementing mDOT programs for HAART in resource-limited settings. In this descriptive study, we used a mixed-methods approach to describe the process of implementing mDOT for an ongoing randomized control trial (RCT) in Beira, Mozambique. Interviews with clinic staff, mDOT peers, and participants provided information on design elements, problems with implementation, satisfaction, and benefits. Acceptability and feasibility measures were obtained from the RCT. Most (81%, N = 350) eligible persons agreed to participate, and of those randomized to mDOT (n = 174), 95% reported that their time with peers was beneficial. On average, participants kept 93% of the 30 required daily mDOT visits. Key components of the intervention's success included using peers who were well accepted by clinic staff, adequate training and retention of peers, adapting daily visit requirements to participants' work schedules and physical conditions, and reimbursing costs of transportation. This study identified aspects of mDOT that are effective and can be adopted by other clinics treating HIV patients. PMID- 17133198 TI - Development and assessment of an innovative culturally sensitive educational videotape to improve adherence to highly active antiretroviral therapy in Soweto, South Africa. AB - The increasing availability of antiretroviral medication (ARV) therapy in the face of limited chronic medication-taking experience among resource-poor South Africans has raised concerns about adequate adherence to these medications. We hypothesized that a culturally sensitive audiovisual patient education program would be of substantial and measurable benefit in increasing patient understanding of the concepts of ARV resistance risk and medication-taking skills. To identify potential barriers to adherence and successful strategies to promote adherence, 6 focus groups with health care providers and HIV-positive adherence counselors were held, resulting in the production of a 17-minute culturally sensitive educational videotape. Basic drug-taking concepts and practical advice on how to improve adherence were presented in the videotape. Thirty-four HIV-positive patients (including 11 ARV-naive patients and 23 ARV experienced patients) were shown the educational videotape, and their knowledge about medication taking was evaluated by a 24-point pre- and postvideotape questionnaire. On average, the 34 patients gained 2.2 knowledge points (P = 0.021). ARV-naive patients had an average improvement of 3.0 points (P = 0.0028), with most significant gains in the areas of understanding medication-taking strategies and side effects. These preliminary findings indicate that a culturally sensitive educational videotape can improve medication-taking knowledge in South Africa and that further study of the potential efficacy of using media technology to improve individuals' adherence to ARV therapy is warranted. PMID- 17133199 TI - The elusive gold standard. Future perspectives for HIV adherence assessment and intervention. AB - There is no "gold standard" for the assessment of adherence to HIV/AIDS medications. Similarly, there is no single optimal tool that enhances adherence to HIV/AIDS treatment regimens. This article presents a model that provides a heuristic for selecting adherence assessment approaches and intervention strategies based on the purpose for which each is to be used. First, a broad distinction is made between research and clinical settings. Second, with each of these settings, the selection of assessments and interventions is based on the extent to which the focus is on HIV/AIDS in general or on adherence in particular. Examples applying the model are provided. Finally, new dimensions are discussed for expanding the model, with particular attention to applying the model to the resource-limited settings that are so important in efforts to reduce the morbidity and mortality associated with the global threat of HIV/AIDS. PMID- 17133200 TI - Examining theoretic models of adherence for validity in resource-limited settings. A heuristic approach. AB - Mortality and morbidity for HIV-infected patients in resource-limited settings have declined markedly with increased access to combination antiretroviral therapy. One explanation for this is the high rates of adherence that patients in these settings have achieved. To help patients sustain early successes, detailed understanding of adherence processes in these locales is needed. This understanding is best arrived at using socioculturally valid theoretic models as a guide. This article introduces a heuristic schema for examining the validity of conceptual models of adherence in resource-limited settings. The schema consists of 4 analytic questions to be asked of a model being considered for use in a sociocultural context other than the one in which it was developed. To demonstrate use of the schema, an illustrative validation exercise in which the 4 questions are applied to the information-motivation-behavioral skills model of behavioral change is carried out. Examples for the illustrative exercise come from preliminary qualitative research in HIV/AIDS treatment settings in Mbarara, Uganda and Jos, Nigeria. PMID- 17133201 TI - Efficacy of interventions in improving highly active antiretroviral therapy adherence and HIV-1 RNA viral load. A meta-analytic review of randomized controlled trials. AB - Adherence to highly active antiretroviral therapy (HAART) is generally suboptimal, limiting the effectiveness of HAART. This meta-analytic review examined whether behavioral interventions addressing HAART adherence are successful in increasing the likelihood of a patient attaining 95% adherence or an undetectable HIV-1 RNA viral load (VL). We searched electronic databases from January 1996 to September 2005, consulted with experts in the field, and hand searched reference sections from relevant articles. Nineteen studies (with a total of 1839 participants) met the selection criteria of describing a randomized controlled trial among adults evaluating a behavioral intervention with HAART adherence or VL as an outcome. Random-effects models indicated that across studies, participants in the intervention arm were more likely than those in the control arm to achieve 95% adherence (odds ratio [OR] = 1.50, 95% confidence interval [CI]: 1.16 to 1.94); the effect was nearly significant for undetectable VL (OR = 1.25; 95% CI: 0.99 to 1.59). The intervention effect for 95% adherence was significantly stronger in studies that used recall periods of 2 weeks or 1 month (vs. /=1.0 log10 reduction in HIV-1 RNA level. A second analysis consisted of linear regression assessing the effect of covariates on log10 HIV-1 RNA reduction from baseline to 6 months. RESULTS: In multivariate analyses, achieving virologic success at 6 months was associated with high medical services utilization [adjusted odds ratio [AOR] = 10.0 (1.4, 73.9); P = 0.02] and with the use of case management services [AOR = 5.8 (1.1, 30.5); P = 0.04]. Both services resulted in a larger reduction in log10 HIV-1 RNA from baseline (difference in slopes: -0.9 and -1.0, respectively; P = 0.02 for both). Referral to off-site substance abuse services treatment did not significantly predict either virologic outcome. CONCLUSIONS: Among individuals who receive DAART, the utilization of on-site medical and case management services was independently associated with improved virologic outcomes. These results suggest the potential utility of integrating these services into DAART interventions (DAART-Plus) targeting HIV-infected drug users with problematic adherence. PMID- 17133205 TI - How qualitative methods contribute to understanding combination antiretroviral therapy adherence. AB - Strict adherence to medication regimens is generally required to obtain optimal response to combination antiretroviral therapy (ART). Yet, we have made limited progress in developing strategies to decrease the prevalence of nonadherence. As we work to understand adherence in developed countries, the introduction of ART in resource-poor settings raises novel challenges. Qualitative research is a scientific approach that uses methods such as observation, interviews, and verbal interactions to gather rich in-depth information about how something is experienced. It seeks to understand the beliefs, values, and processes underlying behavioral patterns. Qualitative methods provide powerful tools for understanding adherence. Culture-specific influences, medication beliefs, access, stigma, reasons for nonadherence, patterns of medication taking, and intervention fidelity and measurement development are areas ripe for qualitative inquiry. A disregard for the social and cultural context of adherence or the imposition of adherence models inconsistent with local values and practices is likely to produce irrelevant or ineffective interventions. Qualitative methods remain underused in adherence research. We review appropriate qualitative methods for and provide an overview of the qualitative research on ART nonadherence. We discuss the rationales for using qualitative methods, present 2 case examples illustrating their use, and discuss possible institutional barriers to their acceptance. PMID- 17133206 TI - Moving from theory to research to practice. Implementing an effective dyadic intervention to improve antiretroviral adherence for clinic patients. AB - There is a dearth of evidence on the relative efficacy of intervention modalities to improve and maintain patient adherence to antiretroviral medications. Although empiric findings from research on HIV/AIDS, other diseases, and chronic medical conditions consistently demonstrate that social support plays an important role in facilitating adherence, few HIV/AIDS interventions have directly targeted this factor. Ewart's social action theory emphasizes the role of social relationships in behavior change and provides a comprehensive and useful guide to the development of interventions for adherence. We describe the development, content, and testing of SMART Couples, an effective antiretroviral adherence intervention that is grounded in social action theory and designed to enhance social support for ART adherence. Finally, we discuss some of the challenges of translating findings from the randomized clinical trial of this intervention into clinical practice and offer recommendations for integration of lessons learned into ongoing clinical care. PMID- 17133207 TI - Practical and conceptual challenges in measuring antiretroviral adherence. AB - Accurate measurement of antiretroviral adherence is essential for targeting and rigorously evaluating interventions to improve adherence and prevent viral resistance. Across diseases, medication adherence is an individual, complex, and dynamic human behavior that presents unique measurement challenges. Measurement of medication adherence is further complicated by the diversity of available measures, which have different utility in clinical and research settings. Limited understanding of how to optimize existing adherence measures has hindered progress in adherence research in HIV and other diseases. Although self-report is the most widely used adherence measure and the most promising for use in clinical care and resource-limited settings, adherence researchers have yet to develop evidence-based standards for self-reported adherence. In addition, the use of objective measures, such as electronic drug monitoring or pill counts, is limited by poor understanding of the source and magnitude of error biasing these measures. To address these limitations, research is needed to evaluate methods of combining information from different measures. The goals of this review are to describe the state of the science of adherence measurement, to discuss the advantages and disadvantages of common adherence measurement methods, and to recommend directions for improving antiretroviral adherence measurement in research and clinical care. PMID- 17133208 TI - Adjusting and censoring electronic monitoring device data. Implications for study outcomes. AB - Electronic monitoring device (EMD) data are widely used to measure adherence in HIV medication adherence research. EMD data represent an objective measure of adherence and arguably provide more valid data than other methods such as self reported measures, pill counts, and drug level concentration. Moreover, EMD data are longitudinal, include many measurements, and yield a rich data set. This article illustrates potential pitfalls associated with this measurement technique, including lack of clarity associated with EMD data, and the extent to which adherence outcomes are affected by data management decisions. Recommendations are given regarding what information should be included in publications that report results based on EMD data so as to facilitate comparisons between studies. PMID- 17133209 TI - Assessing the effectiveness of antiretroviral adherence interventions. Using marginal structural models to replicate the findings of randomized controlled trials. AB - Randomized controlled trials of interventions to improve adherence to antiretroviral medications are not always feasible. Marginal structural models (MSM) are a statistical methodology that aims to replicate the findings of randomized controlled trials using observational data. Under the assumption of no unmeasured confounders, 3 MSM estimators are available to estimate the causal effect of an intervention. Two of these estimators, G-computation and inverse probability of treatment weighted (IPTW), can be implemented using standard software. G-computation relies on fitting a multivariable regression of adherence on the intervention and confounders. Thus, it is related to the standard multivariable regression approach to estimating causal effects. In contrast, IPTW relies on fitting a multivariable logistic regression of the intervention on confounders. This article reviews the implementation of these methods, the assumptions underlying them, and interpretation of results. Findings are illustrated with a theoretic data example in which MSM are used to estimate the effect of a behavioral intervention on adherence to antiretroviral therapy. PMID- 17133210 TI - Covering all the bases: targeting HIV-1 integrase. PMID- 17133211 TI - Antiretroviral activity, pharmacokinetics, and tolerability of MK-0518, a novel inhibitor of HIV-1 integrase, dosed as monotherapy for 10 days in treatment-naive HIV-1-infected individuals. AB - BACKGROUND: MK-0518 is a novel HIV-1 integrase strand transfer inhibitor with potent in vitro activity against HIV-1 (95% inhibitory concentration [IC95] = 33 nM in 50% human serum) and good bioavailability in uninfected subjects. This study explored the antiretroviral activity and safety of MK-0518 versus placebo for 10 days as monotherapy in antiretroviral therapy-naive HIV-1-infected patients with plasma HIV-1 RNA levels of at least 5000 copies/mL and CD4 T-cell counts of at least 100 cells/mm. METHODS: This was a multicenter, double-blind, randomized, placebo-controlled 2-part study, with the first part using MK-0518 in 1 of 4 doses (100, 200, 400, and 600 mg) versus placebo (randomized 1:1:1:1:1) given twice daily for 10 days of monotherapy. Patients were monitored for safety, pharmacokinetic parameters, and antiretroviral effect. RESULTS: Thirty-five patients were enrolled (6-8 patients per treatment group) and completed 10 days of therapy; the mean baseline log10 HIV RNA level ranged from 4.5 to 5.0 copies/mL in each group. On day 10, the mean decrease from baseline in the log10 HIV RNA level was -0.2 copies/mL for the placebo group and -1.9, -2.0, -1.7 and 2.2 log10 copies/mL for the MK-0518 100-, 200-, 400-, and 600-mg treatment groups, respectively. All dose groups had superior antiretroviral activity compared with placebo (P < 0.001 for comparison of each dose with placebo). At least 50% of patients in each MK-0518 dose group achieved an HIV RNA level <400 copies/mL by day 10. Mean trough MK-0518 concentrations at each dose exceeded the IC95 of 33 nM. Study therapy was generally well tolerated. The most common adverse experiences were headache and dizziness; these were similar between active and control groups. There were no discontinuations because of adverse experiences and no serious adverse experiences. CONCLUSIONS: MK-0518 showed potent antiretroviral activity as short-term monotherapy and was generally well tolerated at all doses. Based on these results, part 2 of the study, a dose ranging 48-week trial of MK-0518 versus efavirenz in a combination regimen, has been initiated. PMID- 17133212 TI - Frequency of gynecologic follow-up and cervical cancer screening in the Swiss HIV cohort study. AB - BACKGROUND: According to current recommendations, HIV-infected women should have at least 1 gynecologic examination per year. OBJECTIVES: To analyze factors associated with frequency of gynecologic follow-up and cervical cancer screening among HIV-infected women followed in the Swiss HIV Cohort Study (SHCS). METHODS: Half-yearly questionnaires between April 2001 and December 2004. At every follow up visit, the women were asked if they had had a gynecologic examination and a cervical smear since their last visit. Longitudinal models were fitted with these variables as outcomes. RESULTS: A total of 2186 women were included in the analysis. Of the 1146 women with complete follow-up in the SHCS, 35.3% had a gynecologic examination in each time period, whereas 7.4% had never gone to a gynecologist. Factors associated with a poor gynecologic follow-up were older age, nonwhite ethnicity, less education, underweight, obesity, being sexually inactive, intravenous drug use, smoking, having a private infectious disease specialist as a care provider, HIV viral load <400 copies/mL, and no previous cervical dysplasia. No association was seen for living alone, CD4 cell count, and positive serology for syphilis. CONCLUSIONS: Gynecologic care among well-followed HIV-positive women is poor and needs to be improved. PMID- 17133213 TI - Tipranavir without ritonavir does not acutely induce peripheral insulin resistance in a rodent model. PMID- 17133214 TI - Scaling-up highly active antiretroviral therapy (HAART) in Peru: problems on the horizon. PMID- 17133215 TI - Renal function in patients receiving tenofovir with ritonavir/lopinavir or ritonavir/atazanavir in the HIV Outpatient Study (HOPS) cohort. PMID- 17133216 TI - Between genotype and phenotype. AB - Genome-wide profiling of cytosine methylation for three human chromosomes in a variety of normal tissues provides a reference resource to begin to decode the epigenetic regulation of gene expression in normal development and to understand gene silencing in cancer. PMID- 17133218 TI - Toward a human epigenome. PMID- 17133219 TI - A new piece in the nephrotic puzzle. PMID- 17133220 TI - Making the commitment to meiosis. PMID- 17133221 TI - New insights into the biological basis of genomic disorders. PMID- 17133224 TI - The cadherin superfamily in neuronal connections and interactions. AB - Neural development and the organization of complex neuronal circuits involve a number of processes that require cell-cell interaction. During these processes, axons choose specific partners for synapse formation and dendrites elaborate arborizations by interacting with other dendrites. The cadherin superfamily is a group of cell surface receptors that is comprised of more than 100 members. The molecular structures and diversity within this family suggest that these molecules regulate the contacts or signalling between neurons in a variety of ways. In this review I discuss the roles of three subfamilies - classic cadherins, Flamingo/CELSRs and protocadherins - in the regulation of neuronal recognition and connectivity. PMID- 17133225 TI - High glycogen levels in the hippocampus of patients with epilepsy. AB - During intense cerebral activation approximately half of the glucose plus lactate taken up by the human brain is not oxidized and could replenish glycogen deposits, but the human brain glycogen concentration is unknown. In patients with temporal lobe epilepsy, undergoing curative surgery, brain biopsies were obtained from pathologic hippocampus (n=19) and from apparently 'normal' cortical grey and white matter. We determined the in vivo brain glycogen level and the activity of glycogen phosphorylase and synthase. Regional differences in glycogen concentration were examined similarly in healthy pigs (n=5). In the patients, the glycogen concentration in 'normal' grey and white matter was 5 to 6 mmol/L, but much higher in the hippocampus, 13.1+/-4.3 mmol/L (mean+/-s.d.; P<0.001); the activities of glycogen phosphorylase and synthase displayed the same pattern. In normal hippocampus from pigs, glycogen was similarly higher than in grey and white matter. Consequently, in human grey and white matter and, particularly, in the hippocampus of patients with temporal lope epilepsy, glycogen constitutes a large, active energy reserve, which may be of importance for energy provision during sustained synaptic activity as epileptic seizures. PMID- 17133226 TI - Detection of the ischemic penumbra using pH-weighted MRI. AB - The classic definition of the ischemic penumbra is a hypoperfused region in which metabolism is impaired, but still sufficient to maintain cellular polarization. Perfusion- and diffusion-weighted MRI (PWI, DWI) can identify regions of reduced perfusion and cellular depolarization, respectively, but it often remains unclear whether a PWI-DWI mismatch corresponds to benign oligemia or a true penumbra. We hypothesized that pH-weighted MRI (pHWI) can subdivide the PWI-DWI mismatch into these regions. Twenty-one rats underwent permanent middle cerebral artery occlusion and ischemic evolution over the first 3.5 h post-occlusion was studied using multiparametric MRI. End point was the stroke area defined by T(2) hyperintensity at 24 h. In the acute phase, areas of reduced pH were always larger than or equal to DWI deficits and smaller than or equal to PWI deficits. Group analysis showed that pHWI deficits during this phase coincided with the resulting infarct area at endpoint. Final infarcts were smaller than PWI deficits (range 65% to 90%, depending on the severity of the occlusion) and much larger than acute DWI deficits. These data suggest that the outer boundary of the hypoperfused area showing a decrease in pH without DWI abnormality may correspond to the outer boundary of the ischemic penumbra, while the hypoperfused region at normal pH may correspond to benign oligemia. These first results show that pHWI can provide information complementary to PWI and DWI in the delineation of ischemic tissue. PMID- 17133227 TI - Activin is a neuronal survival factor that is rapidly increased after transient cerebral ischemia and hypoxia in mice. AB - One approach for developing targeted stroke therapies is to identify the neuronal protective and destructive signaling pathways and gene expression that follow ischemic insult. In some neural injury models, the transforming growth factor beta family member activin can provide neuroprotective effects in vivo and promote neuronal survival. This study tests if activin supports cortical neurons after ischemic challenge in vitro and if signals after cerebral ischemia involve activin in vivo. In a defined cell culture model that uses hydrogen peroxide (H(2)O(2))-free radical stress, activin addition maintained neuronal survival. H(2)O(2) treatment increased activin mRNA twofold in surviving cortical neurons, and inhibition of activin with neutralizing antibodies caused neuronal death. These data identify activin gene changes as a rapid response to oxidative stress, and indicate that endogenous activin acts as a protective factor for cortical neurons in vitro. Similarly, after transient focal cerebral ischemia in adult mice, activin mRNA increased at 1 and 4 h ipsilateral to the infarct but returned to control values at 24 h after reperfusion. Intracellular activated smad signals were detected in neurons adjacent to the infarct. Activin was also increased after 2 h of 11% hypoxia. Activin mRNA increased at 1 h but not 4 or 24 h after hypoxia, similar to the time course of erythropoietin and vascular endothelial growth factor induction. These findings identify activin as an early-regulated gene response to transient ischemia and hypoxia, and its function in cortical neuron survival during oxidative challenge provides a basis to test activin as a potential therapeutic in stroke injury. PMID- 17133228 TI - Noninvasive and three-dimensional imaging of CMRO(2) in rats at 9.4 T: reproducibility test and normothermia/hypothermia comparison study. AB - Ability to image cerebral metabolic rate of oxygen (CMRO(2)) is essential for studying the fundamental role of oxidative metabolism in brain function and disease. We have demonstrated recently that three-dimensional (3D) CMRO(2) images can be obtained in the rat brain during a 2-min (17)O(2) inhalation using the (17)O MR spectroscopic imaging (MRSI) approach at high field. The feasibility for establishing a completely noninvasive approach for imaging CMRO(2) has also been demonstrated. In this study, we further explored the feasibility of (17)O MRSI approach for performing repeated CMRO(2) measurements within a short period of time and evaluated the reproducibility of the repeated measurements. Subsequently, we applied the (17)O MRSI approach to measure CMRO(2) and cerebral blood flow (CBF) values at two brain temperatures in the alpha-chloralose anesthetized rat brain at 9.4 T. Finally, we tested the validity of simplified model for noninvasively determining CMRO(2) in normothermic and hypothermic rat brain. The results show (i) an excellent reproducibility among repeated measurements of 3D CMRO(2) images under the same physiologic condition; (ii) a 44% decrease of CMRO(2) across the rat brain at mild hypothermic (32 degrees C) condition as compared with normothermic (37 degrees C) condition; and (iii) a close correlation between CMRO(2) and CBF within a relatively wide physiologic range. This study demonstrates the capability of (17)O MRSI approach for noninvasively imaging CMRO(2) and its changes caused by physiologic perturbation. This approach, thus, should provide a promising neuroimaging modality for studying oxidative metabolism and bioenergetics associated with brain functions and diseases. PMID- 17133229 TI - Propellers of growth trajectories to obesity and the metabolic syndrome. PMID- 17133230 TI - Early adiposity rebound: causes and consequences for obesity in children and adults. AB - Childhood obesity is an important public health problem, with a rapidly increasing frequency worldwide. Identification of critical periods for the development of childhood and adolescent obesity could be very useful for targeting prevention measures. Weight status in early childhood is a poor predictor of adult adiposity status, and most obese adults were not obese as children. We first proposed to use the body mass index (BMI) charts to monitor individual BMI development. The adiposity rebound (AR) corresponds to the second rise in BMI curve that occurs between ages 5 and 7 years. It is not as direct a measure as BMI at any age, but because it involves the examination of several points during growth, and because it is identified at a time when adiposity level clearly change directions, this method provides information that can help us understand individual changes and the development of health risks. An early AR is associated with an increased risk of overweight. It is inversely associated with bone age, and reflects accelerated growth. The early AR recorded in most obese subjects and the striking difference in the mean age at AR between obese subjects (3 years) and non-obese subjects (6 years) suggest that factors have operated very early in life. The typical pattern associated with an early AR is a low BMI followed by increased BMI level after the rebound. This pattern is recorded in children of recent generations as compared to those of previous generations. This is owing to the trend of a steeper increase of height as compared to weight in the first years of life. This typical BMI pattern (low, followed by high body fatness level) is associated with metabolic diseases such as diabetes and coronary heart diseases. Low body fatness before the AR suggests that an energy deficit had occurred at an early stage of growth. It can be attributable to the high-protein, low-fat diet fed to infants at a time of high energy needs, the former triggering height velocity and the latter decreasing the energy density of the diet and then reducing energy intake. The high-fat, low-protein content of human milk may contribute to its beneficial effects on growth processes. Early (pre- and postnatal) life is a critical period during which environmental factors may programme adaptive mechanisms that will persist in adulthood. Under-nutrition in fetal life or during the first years after birth may programme a thrifty metabolism that will exert adverse effects later in life, especially if the growing child is exposed to overnutrition. These observations stress the importance of an adequate nutritional status in childhood and the necessity to provide nutritional intakes adapted to nutritional needs at various stages of growth. Because the AR reflects particular BMI patterns, it is a useful tool for the paediatrician to monitor the child's adiposity development and for researchers to investigate the different developmental patterns leading to overweight. It contributes to the understanding of chronic disease programming and suggests new approaches to obesity prevention. PMID- 17133231 TI - Early growth, and coronary heart disease and type 2 diabetes: experiences from the Helsinki Birth Cohort Studies. AB - Experiences during critical periods of development through the mechanisms of programming have consequences on later health outcomes. Observations worldwide linking a small body size at birth with adult health outcomes have greatly added to our understanding of the early origins of several non-communicable diseases like coronary heart disease (CHD) and type 2 diabetes. The pattern of growth predisposing to CHD is characterized by a small body size at birth and thinness through infancy up to 2 years of age, followed by accelerated gain in weight and body mass index (BMI) later in childhood. The early growth patterns of individuals who later develop type 2 diabetes very much resemble the growth patterns of CHD, that is, a small body size at birth and thinness at 1 year of age followed by higher body mass indices later in childhood. Numerous studies support the importance of events during critical periods of growth in the pathogenesis of many non-communicable diseases like CHD and type 2 diabetes. There are several possible mechanisms through which a non-optimal early growth associated with accelerated weight gain in childhood could lead to these diseases. To get a better understanding of the patterns of growth affecting adult health outcomes, a life-course approach to the development of chronic diseases needs to be taken. PMID- 17133232 TI - The thrifty 'catch-up fat' phenotype: its impact on insulin sensitivity during growth trajectories to obesity and metabolic syndrome. AB - The analyses of large epidemiological databases have suggested that infants and children who show catch-up growth, or adiposity rebound at a younger age, are predisposed to the development of obesity, type 2 diabetes and cardiovascular diseases later in life. The pathophysiological mechanisms by which these growth trajectories confer increased risks for these diseases are obscure, but there is compelling evidence that the dynamic process of catch-up growth per se, which often overlaps with adiposity rebound at a younger age, is characterized by hyperinsulinemia and by a disproportionately higher rate in the recovery of body fat than lean tissue (i.e. preferential 'catch-up fat'). This paper first focuses upon the almost ubiquitous nature of this preferential 'catch-up fat' phenotype across the life cycle as a risk factor for obesity and insulin-related complications - not only in infants and children who experienced catch-up growth after earlier fetal or neonatal growth retardation, or after preterm birth, but also in adults who show weight recovery after substantial weight loss owing to famine, disease-cachexia or periodic dieting. It subsequently reviews the evidence indicating that such preferential catch-up fat is primarily driven by energy conservation (thrifty) mechanisms operating via suppressed thermogenesis, with glucose thus spared from oxidation in skeletal muscle being directed towards de novo lipogenesis and storage in white adipose tissue. A molecular physiological framework is presented which integrates emerging insights into the mechanisms by which this thrifty 'catch-up fat' phenotype crosslinks with early development of insulin and leptin resistance. In the complex interactions between genetic constitution of the individual, programming earlier in life, and a subsequent lifestyle of energy dense foods and low physical activity, this thrifty 'catch-up fat' phenotype--which probably evolved to increase survival capacity in a hunter-gatherer lifestyle of periodic food shortages--is a central event in growth trajectories to obesity and to diseases that cluster into the insulin resistance (metabolic) syndrome. PMID- 17133233 TI - Adiposity in children born small for gestational age. AB - Epidemiological studies indicate that children born small for gestational age (SGA) have an increased risk of metabolic and cardiovascular disorders as adults. This suggests that foetal undernutrition leads to permanent metabolic alterations, which predispose to metabolic abnormalities upon exposure to environmental factors such as low physical activity and/or high-energy intake in later life (thrifty phenotype hypothesis). However, this relationship is not restricted to foetal undernutrition or intrauterine growth retardation, but is also found for children born premature, or for high birth weight children. Furthermore, early post-natal nutrition, and more specifically catch-up growth, appear to modulate cardiovascular risk as well. Intrauterine growth retardation can be induced in animal models by energy/protein restriction, or ligation of uterine arteries. In such models, altered glucose homeostasis, including low beta cell mass, low insulin secretion and insulin resistance is observed after a few weeks of age. In humans, several studies have confirmed that children born SGA have insulin resistance as adolescents and young adults. Alterations of glucose homeostasis and increased lipid oxidation can indeed be observed already in non diabetic children born SGA at early pubertal stages. These children also have alterations of stature and changes in body composition (increased fat mass), which may contribute to the pathogenesis of insulin resistance. Permanent metabolic changes induced by foetal/early neonatal nutrition (metabolic inprinting) may involve modulation of gene expression through DNA methylation, or alterations of organ structure. It is also possible that events occurring during foetal/neonatal development lead to long-lasting alterations of the hypothalamo pituitary-adrenal axis or the hypothalamo-pituitary-insulin-like growth factor-1 axis. PMID- 17133234 TI - Obesity during growth in Switzerland: role of early socio-cultural factors favouring sedentary activities. AB - Depending upon age, gender and geographical area, 3-20% of the children and young adolescents in Switzerland are overweight and 0-6% obese, using the criteria of the International Obesity Task Force. The most likely explanation for this increasing prevalence of overweight and obesity is a decline in physical activity, and hence diminished energy expenditure that is not matched by a corresponding reduction in energy intake. In this paper, we first review the epidemiological situation in Switzerland regarding the rising prevalence of obesity and the decline in physical activity, and then focus upon the environmental, social and cultural factors that predispose children to sedentary behaviours. Several of these socio-cultural factors and in particular television viewing and poor-parental model, confer early 'learned behaviours' for low physical activity which track throughout growth into adulthood, and which also predispose to the overconsumption of less healthy foods. It is time to focus on preventive strategies directed at curtailing these 'learned behaviours', that we have acquired during the transition from the stone-age to the chip-age, if we want to hold the current pandemic of obesity. PMID- 17133235 TI - Developmental origins of obesity: a sympathoadrenal perspective. AB - Environmental exposures at crucial points in development permanently alter sympathoadrenal function in mammals. Both the sympathetic innervation of peripheral tissues and the responsiveness of sympathetic nerves and adrenal medulla to standard stimuli are susceptible to modification by exposures in early life. Several conditions studied in the laboratory, including environmental temperature, litter size and maternal nutrition, in addition to affecting sympathoadrenal function also produce larger, fatter offspring, raising the possibility that developmental programming of the sympathetic nervous system (SNS) may contribute to acquisition of an obese phenotype. The specific changes noted in all three circumstances include evidence of an increase in sympathetic innervation in pancreas and retroperitoneal fat. By contrast, SNS development is impaired in experimental models of intrauterine growth retardation. Although the physiological implications of increased sympathetic innervation in pancreas and retroperitoneal fat are not fully understood, these changes seen in animals reared at cool temperatures, in small litters or by mothers fed refined carbohydrate diets likely reflect an early enhancement of the offspring's capacity to take up and store glucose. If so, the tendency of these animals to gain weight and accumulate fat may represent an adaptive response to 'over nutrition' in early life. PMID- 17133236 TI - Dietary fat and fat types as early determinants of childhood obesity: a reappraisal. AB - BACKGROUND: The growing prevalence of childhood overweight and obesity has renewed interest in determining the influence of the maternal and infant diet on the risk of developing excess fat mass later in life. APPROACH: Review of available human and animal data reporting the effects of dietary fat and fat types early in life on adipose development. RESULTS: Rodent studies tend to show that maternal high-fat feeding during pregnancy and lactation results in increased adiposity of the offspring. Nevertheless, today there is a lack of population-based studies investigating this potential detrimental effect of maternal high-fat intake. Most epidemiological studies, performed so far, do not find any association between the level of dietary fat intake of infants and children and body weight and/or fatness. Regarding fat types exposure to high levels of dietary n-6 fatty acids during gestation and post-natal life, has been shown to promote obesity in mice. Nevertheless, other rodent studies do not demonstrate such an effect. CONCLUSION: There is no evidence supporting a restriction of fat intake during the first two post-natal years but the potential detrimental effects of maternal high-fat intake during gestation should be further investigated. The role of dietary fat types as early determinants of childhood obesity has so far been poorly studied. Robust evidence to support the adipogenic effects of n-6 fatty acids enriched-diets is currently lacking but this hypothesis is of importance and should be further evaluated in different animal models as well as in longitudinal human studies. PMID- 17133237 TI - Weight cycling during growth and beyond as a risk factor for later cardiovascular diseases: the 'repeated overshoot' theory. AB - In people trying to lose weight, there are often repeated cycles of weight loss and regain. Weight cycling is, however, not limited to obese adults but affects people of normal weight, particularly young women, who are unhappy with their appearance. Furthermore, the onset of a pattern of weight cycling is shifting towards younger ages, owing to the increasing prevalence of overweight and obesity in children and adolescents, and the pressure from the media and society for a slim image even for normal weight children. Although there is still controversy whether weight cycling promotes body fat accumulation and obesity, there is mounting evidence from large population studies for increased cardiovascular risks in response to a behavior of weight cycling. Potential mechanisms by which weight cycling contributes to cardiovascular morbidity include hypertension, visceral fat accumulation, changes in adipose tissue fatty acid composition, insulin resistance and dyslipidemia. Moreover, fluctuations in blood pressure, heart rate, sympathetic activity, glomerular filtration rate, blood glucose and lipids that may occur during weight cycling--with overshoots above normal values during weight regain periods--put an additional load on the cardiovascular system, and may be easily overlooked if humans or animals are studied during a state of relatively stable weight. Overshoot of those risks factors, when repeated over time, will stress the cardiovascular system and probably contribute to the overall cardiovascular morbidity of weight cycling. PMID- 17133239 TI - Should clinicians worry about vancomycin-resistant Enterococcus bloodstream infections? PMID- 17133240 TI - Pilot study to test the efficacy and safety of activated recombinant factor VII (NovoSeven) in the treatment of refractory hemorrhagic cystitis following high dose chemotherapy. PMID- 17133241 TI - Overcoming scientific barriers and human prudence. PMID- 17133242 TI - Reply to Drs Caocci and Pisu. PMID- 17133243 TI - A biomarker by any other name... PMID- 17133244 TI - Opioid therapy for the treatment of refractory pain in children with juvenile rheumatoid arthritis. PMID- 17133245 TI - New immune modulatory drugs for systemic lupus erythematosus--what can we expect? PMID- 17133246 TI - Anakinra for the treatment of neonatal-onset multisystem inflammatory disease. PMID- 17133247 TI - Do genetic variations in the adenosine pathway affect patient response to methotrexate? PMID- 17133248 TI - Does diuretic use increase the risk of recurrent gout flares? PMID- 17133249 TI - Antibodies against citrullinated proteins: which test is best? PMID- 17133250 TI - Drug Insight: abatacept for the treatment of rheumatoid arthritis. AB - Although the development of tumor necrosis factor antagonists has improved the clinical outcome of many patients with rheumatoid arthritis (RA), some patients fail to respond to these drugs or have contraindications preventing their use. Other approaches for treating this disease are, therefore, keenly sought. As T cells promote numerous disease pathways in RA, these cells are a logical target for anti-inflammatory therapy. One of the approaches being investigated involves targeting the co-stimulatory signals that accompany antigen-derived signals involved in the activation of T cells. Abatacept is a recombinant fusion protein that interrupts the T-cell co-stimulatory signal mediated through the CD28 CD80/CD86 pathway. Several clinical trials have now confirmed the efficacy of this compound in the treatment of RA. This article discusses the proposed mechanism of action of abatacept and reviews the data from phase II and phase III clinical trials on the safety and efficacy of this drug in RA. PMID- 17133251 TI - Mechanisms of Disease: pathogenesis and treatment of ANCA-associated vasculitides. AB - Wegener's granulomatosis and microscopic polyangiitis are idiopathic systemic vasculitides strongly associated with antineutrophil cytoplasmic autoantibodies (ANCA). In Wegener's granulomatosis, ANCA are mostly directed against proteinase 3 (PR3), whereas in microscopic polyangiitis ANCA are directed against myeloperoxidase; increases in levels of these autoantibodies precede or coincide with clinical relapses in many cases. In vitro, ANCA can further activate primed neutrophils to release reactive oxygen species and lytic enzymes, and, in conjunction with neutrophils, can damage and lyse endothelial cells. Patients with Wegener's granulomatosis or microscopic polyangiitis have an increased percentage of neutrophils that constitutively express PR3 on their membrane. These neutrophils can be stimulated by ANCA, without priming. In vivo, transfer of splenocytes from myeloperoxidase-deficient mice immunized with mouse myeloperoxidase into wild-type mice resulted in pauci-immune systemic vasculitis. A similar experiment in PR3-deficient mice did not cause significant vasculitic lesions. Together, clinical, in vitro and in vivo experimental data support a pathogenic role for ANCA in Wegener's granulomatosis and microscopic polyangiitis, although this role is more evident for myeloperoxidase-specific ANCA than for PR3-specific ANCA. Several controlled trials have led to an evidence-based approach for the treatment of ANCA-associated vasculitis, and further studies, based on new insights into pathogenesis, are in progress. PMID- 17133252 TI - Mechanisms of Disease: genetics of fibromyalgia. AB - Fibromyalgia is characterized by widespread pain and tenderness, and has a significant familial component. The etiology of fibromyalgia remains unclear, but genetic factors seem to have a significant role, and are influenced by environmental factors. Research over the past two decades has demonstrated that genetic polymorphisms in the serotoninergic, dopaminergic and catecholaminergic systems of pain transmission and processing are involved in the etiology of fibromyalgia, but additional candidates continue to emerge. Fibromyalgia is thought to belong to the group of affective spectrum disorders, which include related psychiatric and medical disorders. As the concept of affective spectrum disorders continues to evolve, progress in the understanding of the genetic basis of related functional disorders, such as irritable bowel syndrome and post traumatic-stress disorder, is aiding our understanding of the genetic basis of fibromyalgia. PMID- 17133253 TI - Mechanisms of Disease: the role of immune cells in the pathogenesis of systemic sclerosis. AB - Systemic sclerosis is characterized by extensive fibrosis, microvascular stenosis and autoantibody production. All three characteristics can be accounted for by activation of cells of the immune system. Activation of T cells is antigen-driven and occurs early in the course of the disease, before microscopic evidence of fibrosis. Activated T cells are predominantly of the type 2 T-helper lineage, and produce interleukin-4 and interleukin-13, which induce fibrosis. B cells are also activated early in the course of the disease and, through the production of autoantibodies, cause fibroblasts to adopt a profibrotic phenotype. Macrophages in perivascular infiltrates are activated and produce CC-chemokine ligand 2, transforming growth factor beta and platelet derived growth factor, all of which promote fibrosis and fibroproliferation. These new insights have direct impact on the treatment of patients with systemic sclerosis; therapies that target T cells, B cells and their harmful mediators are a logical approach, and preliminary data are promising. PMID- 17133254 TI - A case of polymyalgia rheumatica, microscopic polyangiitis, and B-cell lymphoma. AB - BACKGROUND: A 73-year-old, previously well woman was admitted to an emergency department because of a 3-month history of severe proximal girdle pain and stiffness with loss of appetite and weight. She was referred to a rheumatologist 10 days after her initial presentation. Within 4 weeks she presented to an outpatient clinic with nausea, vomiting, shortness of breath, painful mouth ulcers, rash on her legs and a further decline in appetite; she was readmitted to hospital. Within 4 months of initial presentation she became jaundiced. INVESTIGATIONS: At initial presentation, physical examination, biochemical, hematological and autoimmune screening, radiography of the pelvis, an abdominal ultrasound, and electromyography were conducted. At referral to a rheumatologist similar tests were repeated. At presentation to the outpatient clinic, hematological and biochemical screening, and a urine dipstick test were conducted. At readmittance to hospital, infectious and autoimmune screening, radiography of the chest, electrocardiogram, ultrasound of the abdomen, and renal biopsy were conducted. At the time of development of jaundice, biochemical and hematological screening, CT of the abdomen and ultrasound-guided biopsy of a pancreatic mass were conducted. DIAGNOSIS: Polymyalgia rheumatica, antineutrophil cytoplasmic-antibody-positive microscopic polyangiitis with renal involvement and B-cell lymphoma of the head of the pancreas. MANAGEMENT: The patient received oral prednisolone 15 mg daily for polymyalgia rheumatica along with alendronate 70 mg weekly. The patient received intravenous cyclophosphamide 500 mg and methylprednisolone 500 mg every 2 weeks for her microscopic polyangiitis with renal involvement. For B-cell lymphoma of the head of the pancreas, the patient received cyclophosphamide, doxorubicin, vincristine and prednisolone once monthly. PMID- 17133256 TI - Preselection of cases through expert clinical and radiological review significantly increases mutation detection rate in multiple epiphyseal dysplasia. AB - Skeletal dysplasias are difficult to diagnose for the nonexpert. In a previous study of patients with multiple epiphyseal dysplasia (MED), we identified cartilage oligomeric matrix protein (COMP) mutations in only 36% of cases and suspected that the low-mutation detection rate was partially due to misdiagnosis. We therefore instituted a clinical-radiographic review system, whereby all cases were evaluated by a panel of skeletal dysplasia experts (European Skeletal Dysplasia Network). Only those patients in whom the diagnosis of MED was confirmed by the panel were screened for mutations. Under this regimen the mutation detection rate increased to 81%. When clinical-radiological diagnostic criteria were relaxed the mutation rate dropped to 67%. We conclude that expert clinical-radiological review can significantly enhance mutation detection rates and should be part of any diagnostic mutation screening protocol for skeletal dysplasias. PMID- 17133257 TI - Using HapMap data: a cautionary note. AB - The HapMap data are being widely used in human genetic studies. We show by direct resequencing of a approximately 6-kb region of chromosome 1 that the HapMap data are unreliable for this region. This region contains a recent mitochondrial (mt) DNA insertion. The HapMap data report the corresponding mtDNA variation and not the nuclear DNA variation. In view of mtDNA insertions of varying lengths throughout the human genome and considerable segmental duplications, it is necessary to use the HapMap data cautiously. PMID- 17133259 TI - Stability of BAT26 in Lynch syndrome colorectal tumours. PMID- 17133260 TI - Multiple QTL influence the serum Lp(a) concentration: a genome-wide linkage screen in the PROCARDIS study. AB - The serum concentration of lipoprotein Lp (a) is known to be highly heritable and associated with cardiovascular risk. A genome-wide variance component linkage analysis was performed to localise quantitative trait loci (QTLs) influencing Lp(a) levels in a large cohort collected in the PROCARDIS coronary heart disease study. Highly significant linkage was detected at the previously described LP(a) locus on chromosome 6q27 (LOD 108). Taking into account the effect of the locus detected on chromosome 6, a highly significant LOD score was detected on chromosome 13q22-31 (LOD 7.0). Another significant region of linkage was observed on chromosomes 11p14-15 (LOD 3.5). The significant peak at 13q22-31 shows an essential overlap with a locus modulating cholesterol in familial hypercholesterolemia. If the gene underlying these loci is the same, it will be a promising candidate target for manipulating LDL-cholesterol and Lp(a). We also detected linkage at a previously identified locus influencing Lp(a) on chromosome 1q23 (LOD 1.5). Our findings provide new and confirmatory information about genomic regions involved in the quantitative variation of Lp(a) and serve as a basis for further studies of candidate genes in these regions. PMID- 17133261 TI - Mental performance in old age dependent on cortisol and genetic variance in the mineralocorticoid and glucocorticoid receptors. AB - Depression and cognitive decline have been associated with changes in circulating cortisol concentrations. Cortisol exerts its functions through mineralocorticoid (MR) and glucocorticoid (GR) receptors. However, data on the influence of variations in the MR and GR genes on depressive symptoms and cognitive functioning in older adults are scarce. Therefore, we explored the impact of MR 215G/C, MR-I180V, GR-ER22/23EK, GR-N363S, and GR-BclI polymorphisms on these end points in the population-based Leiden 85-plus Study. This prospective study includes 563 participants aged 85 years and older, with a mean follow-up of 4.2 years. In this study, high morning cortisol levels (per 1 SD cortisol) associated with impairments in global cognitive functioning (p=0.002) at baseline (age 85). These impairments were mainly attributable to lower attention (p=0.057) and slower processing speed (p=0.014). Similar effects were also observed during follow-up (age 85-90), where participants with higher cortisol levels (per 1 SD cortisol) had impaired global cognitive functioning (p=0.003), as well as impairments in attention (p=0.034) and processing speed (p=0.013). Changes in depressive symptoms were observed for the MR-I180V single-nucleotide polymorphism (SNP), where during follow-up the prevalence of depressive symptoms was higher in the 180V-allele carriers (p=0.049) compared to noncarriers. Dependent on these polymorphisms, no differences in overall and in specific domains of cognitive functioning were observed. In conclusion, the MR-I180V SNP has a specific effect on depressive symptoms, independent from cognitive functioning, and other polymorphisms in the MR and GR genes. In contrast, these genetic variants in the MR and GR genes do not influence cognitive functioning in old age. PMID- 17133262 TI - The effect of intrahippocampal injection of group II and III metobotropic glutamate receptor agonists on anxiety; the role of neuropeptide Y. AB - Earlier studies conducted by our group and by other authors indicated that metabotropic glutamatergic receptor (mGluR) ligands might have anxiolytic activity and that amygdalar neuropeptide Y (NPY) neurons were engaged in that effect. Apart from the amygdala, the hippocampus, another limbic structure, also seems to be engaged in regulation of anxiety. It is rich in mGluRs and contains numerous NPY interneurons. In the present study, we investigated the anxiolytic activity of group II and III mGluR agonists after injection into the hippocampus, and attempted to establish whether hippocampal NPY neurons and receptors were engaged in the observed effects. Male Wistar rats were bilaterally microinjected with the group II mGluR agonist (2S,1'S,2'S)-2-(carboxycyclopropyl)glycine (L-CCG I), group III mGluR agonist O-Phospho-L-serine (L-SOP), NPY, the Y1 receptor antagonist BIBO 3304, and the Y2 receptor antagonist BIIE 0246 into the CA1 or dentate area (DG). The effect of those compounds on anxiety was tested in the elevated plus-maze. Moreover, the effects of L-CCG-I and L-SOP on the expression of NPYmRNA in the hippocampus were studied using in situ hybridization method. It was found that a significant anxiolytic effect was induced by L-SOP injection into the CA1 region or by L-CCG-I injection into the DG. The former effect was inhibited by BIBO 3304, the latter by BIIE 0246. NPY itself showed antianxiety action after injection into both structures. In the CA1 area, the effect of NPY was prevented by BIBO 3304, whereas in the DG by BIIE 0246. Both the mGluR agonists L-CCG-I and L-SOP induced a potent increase in NPYmRNA expression in the DG region of the hippocampal formation. The obtained results indicate that group II and III mGluR agonists, L-CCG-I and L-SOP, as well as NPY display anxiolytic activity in the hippocampus, but act differently in the CA1 and DG. It was observed that group III mGluRs and Y1 receptors were engaged in the response in the CA1 area, whereas group II mGluRs and Y2 receptors played a pivotal role in the DG region. PMID- 17133263 TI - The allosteric potentiation of nicotinic acetylcholine receptors by galantamine ameliorates the cognitive dysfunction in beta amyloid25-35 i.c.v.-injected mice: involvement of dopaminergic systems. AB - Galantamine, a drug for Alzheimer's disease, is a novel cholinergic agent with a dual mode of action, which inhibits acetylcholinesterase and allosterically modulates nicotinic acetylcholine receptors (nAChRs), as a result stimulates catecholamine neurotransmission. In the present study, we investigated whether galantamine exerts cognitive improving effects through the allosteric modulation of nAChR in the intracerebroventricular beta amyloid (Abeta)(25-35)-injected animal model of Alzheimer's disease. Galantamine (3 mg/kg p.o.) significantly increased the extracellular dopamine release in the hippocampus of saline- and Abeta(25-35)-injected mice. The effects of nicotine on the extracellular dopamine release were potentiated by galantamine, but antagonized by mecamylamine, a nAChR antagonist. Abeta(25-35)-injected mice, compared with saline-injected mice, could not discriminate between new and familiar objects in the novel object recognition test and exhibited less freezing response in the fear-conditioning tasks, suggesting Abeta(25-35) induced cognitive impairment. Galantamine improved the Abeta(25-35)-induced cognitive impairment in the novel object recognition and fear-conditioning tasks. These improving effects of galantamine were blocked by the treatment with mecamylamine, SCH-23390, a dopamine-D1 receptor antagonist, and sulpiride, a dopamine-D2 receptor antagonist, but not by scopolamine, a muscarinic acetylcholine receptor antagonist. This study provides the first in vivo evidence that galantamine augments dopaminergic neurotransmission within the hippocampus through the allosteric potentiation of nAChRs. The improving-effects of galantamine on the Abeta(25-35)-induced cognitive impairment may be mediated through the activation of, at least in part, dopaminergic systems, and the enhancement of dopamine release may be one of multiple mechanisms underlying the therapeutic benefit of galantamine. PMID- 17133264 TI - Estrogen receptor beta, but not alpha, mediates estrogen's effect on cocaine induced reinstatement of extinguished cocaine-seeking behavior in ovariectomized female rats. AB - Preclinical and clinical studies indicate that females are more vulnerable to relapse than males, and the neurobiological effects of estrogen are thought to mediate, in part, the sex differences in cocaine-taking behavior. The goal of the present study was to investigate the involvement of estrogen receptor alpha (ERalpha) and beta (ERbeta) in estrogen-mediated increases in cocaine-induced reinstatement of extinguished cocaine-seeking behavior in ovariectomized (OVX) female rats. Rats were initially trained to self-administer cocaine (0.4 mg/kg/inf, i.v.) under a fixed-ratio 1 (FR 1) schedule of reinforcement during daily 2-h sessions. After a 10-day maintenance period, cocaine solutions were replaced with saline, and self-administration was extinguished over a 14-day period. OVX rats were then treated with either the mixed ERalpha/beta agonist estradiol benzoate (EB), the ERalpha-selective agonist, propyl-pyrazole-triol (PPT), the ERbeta-selective agonist, diarylpropionitrile (DPN), or a vehicle control (dimethyl sulfoxide, DMSO). Treatment lasted a total of 9 days, and during this time, rats were assessed for nonreinforced reinstatement of extinguished cocaine-seeking behavior after priming injections of saline or cocaine (5, 10, or 15 mg/kg, i.p.). OVX rats showed no differences in self administration during maintenance or extinction. OVX rats treated with EB exhibited greater responding for cocaine during reinstatement compared to OVX+DMSO controls. Selective activation of ERbeta with DPN also increased cocaine induced reinstatement responding, whereas selective activation of ERalpha with PPT did not affect cocaine-seeking behavior. These results indicate that estrogen influences the propensity for reinstatement of extinguished cocaine-seeking behavior, and that estrogen-mediated enhancement of cocaine-induced reinstatement responding involves the activation of ERbeta. PMID- 17133265 TI - Are some breast cancers sexually transmitted? PMID- 17133266 TI - Three-week schedule of irinotecan plus cisplatin in patients with previously untreated extensive-stage small-cell lung cancer. AB - Irinotecan and cisplatin demonstrated promising outcomes in extensive-stage small cell lung cancer. According to the dosage and schedule of irinotecan, efficacy and toxicity profiles showed subtle differences. This study was designed to evaluate efficacy and toxicity of 3-week schedule of irinotecan/cisplatin in patients with previously untreated extensive-stage small-cell lung cancer. The primary objective was to evaluate response rate and secondary objectives were overall survival and progression-free survival. Patients with previously untreated extensive-stage small-cell lung cancer were enrolled. Irinotecan 65 mg m-2 was administered on days 1 and 8 and cisplatin 60 mg m-2 on day 1. Treatment was repeated every 3 weeks. Seven out of 54 patients (13.0%) had complete response, and partial response was observed in 33 (61.1%). The overall response rate was 74.1% (95% CI; 62.0-82.2%). Stable disease was observed in eight (14.8%) and no progressive disease was observed. After a median follow-up duration of 28.7 months, the median overall survival and progressive-free survival were 13.6 and 6.5 months, respectively. Major grade 3/4 toxicities were neutropenia (50.0%), anorexia (42.6%), diarrhoea (29.6%), fatigue (29.6%) and vomiting (13.0%). There was one treatment-related death owing to pneumonia. Three-week schedule of irinotecan/cisplatin showed effective antitumour activity and moderate toxicities in patients with previously untreated extensive-stage small cell lung cancer. PMID- 17133267 TI - Stage, grade and morphology of tumours of the colon and rectum recorded in the Oxford Cancer Registry, 1995-2003. AB - Data on stage, grade and morphology of 12,761 colorectal cancers registered between 1995 and 2003 by Oxford Cancer Registry are reviewed. Dukes stage is recorded for 81% of colon cancers and for 69% of rectal cancers. Incomplete registry data and changing recording practices may affect future evaluation of bowel cancer screening. PMID- 17133268 TI - Phase I/II study of S-1 combined with paclitaxel in patients with unresectable and/or recurrent advanced gastric cancer. AB - Both paclitaxel and S-1 are effective against gastric cancer, but the optimal regimen for combined chemotherapy with these drugs remains unclear. This phase I/II study was designed to determine the maximum tolerated dose (MTD), recommended dose (RD), dose-limiting toxicity (DLT), and objective response rate of paclitaxel in combination with S-1. S-1 was administered orally at a fixed dose of 80 mg m-2 day-1 from days 1 to 14 of a 28-day cycle. Paclitaxel was given intravenously on days 1, 8, and 15, starting with a dose of 40 mg m-2 day-1. The dose was increased in a stepwise manner to 70 mg m-2. Treatment was repeated every 4 weeks unless disease progression was confirmed. In the phase I portion, 17 patients were enrolled. The MTD of paclitaxel was estimated to be 70 mg m-2 because 40% of the patients given this dose level (two of five) had DLT. The RD was determined to be 60 mg m-2. In the phase II portion, 24 patients, including five with assessable disease who received the RD in the phase I portion, were evaluated. The median number of treatment courses was six (range: 1-17). The incidence of the worst-grade toxicity in patients given the RD was 28 and 8%, respectively. All toxic effects were manageable. The response rate was 54.1%, and the median survival time was 15.5 months. Our phase I/II trial showed that S-1 combined with paclitaxel is effective and well tolerated in patients with advanced gastric cancer. PMID- 17133269 TI - Mutations in BHD and TP53 genes, but not in HNF1beta gene, in a large series of sporadic chromophobe renal cell carcinoma. AB - BHD, TP53, and HNF1beta on chromosome 17 were studied in 92 cases of renal cell carcinoma (46 chromophobe, 19 clear cell, 18 oncocytoma, and nine papillary). Six, thirteen, and zero cases had, respectively BHD, TP53, and HNF1beta mutations, (84% mutations involved chromophobe), suggesting a role for BHD and TP53 in chromophobe subtype. PMID- 17133270 TI - Microarray-based comparative genomic hybridisation of breast cancer patients receiving neoadjuvant chemotherapy. AB - We analysed the molecular genetic profiles of breast cancer samples before and after neoadjuvant chemotherapy with combination doxorubicin and cyclophosphamide (AC). DNA was obtained from microdissected frozen breast core biopsies from 44 patients before chemotherapy. Additional samples were obtained before the second course of chemotherapy (D21) and after the completion of the treatment (surgical specimens) in 17 and 21 patients, respectively. Microarray-based comparative genome hybridisation was performed using a platform containing approximately 5800 bacterial artificial chromosome clones (genome-wide resolution: 0.9 Mb). Analysis of the 44 pretreatment biopsies revealed that losses of 4p, 4q, 5q, 12q13.11 12q13.12, 17p11.2 and 17q11.2; and gains of 1p, 2p, 7q, 9p, 11q, 19p and 19q were significantly associated with oestrogen receptor negativity. 16q21-q22.1 losses were associated with lobular and 8q24 gains with ductal types. Losses of 5q33.3 q4 and 18p11.31 and gains of 6p25.1-p25.2 and Xp11.4 were associated with HER2 amplification. No correlations between DNA copy number changes and clinical response to AC were found. Microarray-based comparative genome hybridisation analysis of matched pretreatment and D21 biopsies failed to identify statistically significant differences, whereas a comparison between matched pretreatment and surgical samples revealed a statistically significant acquired copy number gain on 11p15.2-11p15.5. The modest chemotherapy-driven genomic changes, despite profound loss of cell numbers, suggest that there is little therapeutic selection of resistant non-modal cell lineages. PMID- 17133271 TI - Methylation of tumour suppressor genes APAF-1 and DAPK-1 and in vitro effects of demethylating agents in bladder and kidney cancer. AB - To examine the significance of the methylation level of the p53 target and tumour suppressor genes apoptotic protease activating factor-1 (APAF-1) and death associated protein kinase-1 (DAPK-1) in 80 microdissected tumour samples from transitional cell carcinoma (TCC) of the bladder and 80 tumour samples from clear cell renal cell carcinoma (RCC) as well as from non-tumourous bladder and kidney tissue. Growth-inhibitory effects of the demethylating agents 5-Aza-2' deoxycytidine (5-Aza-CdR) and zebularine were investigated in TCC and RCC cell lines. The methylation frequency of APAF-1 (DAPK-1) was 100% (77%) in TCC and 100% (33%) in RCC. The methylation levels of APAF-1 could differentiate between the individual tumour stages in TCC as well as in RCC. The APAF-1 methylation levels in RCC were significantly higher in tumours larger than 4 cm and in high grade tumours. The methylation frequencies in normal tissue for APAF-1 (DAPK-1) were 11% (8%) in bladder tissue and 9% (5%) in kidney tissue. The growth inhibitory effect of the demethylating agents in TCC (RT4, T24) and RCC (A498, ClearCa-5) cell lines resulted in a 17-132% prolongation of the doubling time (DT). In RCC cell lines, zebularine was superior to 5-Aza-CdR in achieving a DT prolongation. Quantitative real time RT-PCR detected a re-expression of mRNA transcripts of APAF-1 or DAPK-1. In conclusion, demethylating agents effectively retard growth of TCC and RCC cell lines. Methylation level analysis of specific genes has the potential for further tumour characterisation in TCC and RCC. PMID- 17133275 TI - Is there a need to re-examine the approval of bismuth shotshell as a non-toxic alternative to lead based on the precautionary principle? AB - In 1997, the Canadian Wildlife Service (CWS) approved bismuth shotshell as an alternative to lead. We assessed whether there is a need to re-examine the toxicity of bismuth issue using the initial CWS toxicity guideline criteria. Review of the literature suggests that in the context of the precautionary principle, a re-examination of the approval of bismuth shotshell should be undertaken based on animal health issues, associated deficiencies in the analytical data used in the original assessment, and to a lesser extent, human health concerns. PMID- 17133276 TI - An observational case study of ozone and precursors inflow to South East England during an anticyclone. AB - Summertime photochemical air pollution episodes within the United Kingdom have been proposed via modelling studies to be strongly influenced by regional scale inflow of air from the continental European boundary layer. We present a vertically resolved case study using measurements made from the NERC/Met Office BAe 146 research aircraft on 18th August 2005 over the South East of England and the North Sea during a weak anticyclone centred over Northern Europe. The vertical distribution of ozone, CO, NO(x), peroxyacetyl nitrate (PAN) and a wide range of both nonmethane hydrocarbons (NMHC) and oxygenated volatile organic compounds (OVOCs) were determined between 500 ft (approximately 152 m) and 7000 ft (approximately 2134 m) over the East Anglia coastline and 50 km inland. In excess of 80 ppbV ozone was observed within inflowing boundary layer air over the North Sea coast in a broad N-S sloping feature around 60 km wide. The inflowing feature of European origin was also observed further inland within the boundary layer albeit with lower, more variable, ozone mixing ratios. The increased variability in ozone over land was a product of titration by fresh surface emissions of NO via rapid upward transport in thermals, a hypothesis supported by the observed vertical wind speed component. Fast boundary layer mixing over land was further illustrated by a uniform distribution in reactive alkenes. A comparison between aircraft and surface O(3) UK AUN (Automatic Urban Network) measurements showed good agreement with the inland site, Sibton, but marked differences with the coastal monitoring site at Weybourne, potentially due to gradients established by ocean deposition in stably stratified marine air. PMID- 17133272 TI - NFV, an HIV-1 protease inhibitor, induces growth arrest, reduced Akt signalling, apoptosis and docetaxel sensitisation in NSCLC cell lines. AB - HIV-1 protease inhibitor (PI), nelfinavir (NFV) induced growth arrest and apoptosis of NCI-H460 and -H520, A549, EBC-1 and ABC-1 non-small-cell lung cancer (NSCLC) cells in association with upregulation of p21waf1, p27kip1 and p53, and downregulation of Bcl-2 and matrix metalloproteinase (MMP)-2 proteins. We found that NFV blocked Akt signalling in these cells as measured by Akt kinase assay with glycogen synthase kinase-3alpha/beta (GSK-3alpha/beta) as a substrate. To explore the role of Akt signalling in NFV-mediated growth inhibition of NSCLC cells, we blocked this signal pathway by transfection of Akt small interfering RNA (siRNA) in these cells; transient transfection of Akt siRNA in NCI-H460 cells decreased the level of Bcl-2 protein and slowed their proliferation compared to the nonspecific siRNA-transfected cells. Conversely, forced-expression of Akt partially reversed NFV-mediated growth inhibition of these cells, suggesting that Akt may be a molecular target of NFV in NSCLC cells. Also, we found that inhibition of Akt signalling by NFV enhanced the ability of docetaxel to inhibit the growth of NCI-H460 and -H520 cells, as measured by MTT assay. Importantly, NFV slowed the proliferation and induced apoptosis of NCI-H460 cells present as tumour xenografts in nude mice without adverse systemic effects. Taken together, this family of compounds might be useful for the treatment of individuals with NSCLC. PMID- 17133277 TI - Observations of urban airborne particle number concentrations during rush-hour conditions: analysis of the number based size distributions and modal parameters. AB - A summertime study of the number concentration and the size distribution of combustion derived nanometre sized particles (termed nanoparticles) from diesel and spark-ignition (SI) engine emissions were made under rush-hour and free-flow traffic conditions at an urban roadside location in Leeds, UK in July 2003. The measured total particle number concentrations (N(TOTAL)) were of the order 1.8 x 10(4) to 3.4 x 10(4) cm(-3), and tended to follow the diurnal traffic flow patterns. The N(TOTAL) was dominated by particles < or =100 nm in diameter which accounted for between 89-93% of the measured particle number. By use of a log normal fitting procedure, the modal parameters of the number based particle size distribution of urban airborne particulates were derived from the roadside measurements. Four component modes were identified. Two nucleation modes were found, with a smaller, more minor, mode composed principally of sub-11 nm particles, believed to be derived from particles formed from the nucleation of gaseous species in the atmosphere. A second mode, much larger in terms of number, was composed of particles within the size range of 10-20 nm. This second mode was believed to be principally derived from the condensation of the unburned fuel and lube oil (the solvent organic fraction or SOF) as it cooled on leaving the engine exhaust. Third and fourth modes were noted within the size ranges of 28-65 nm and 100-160 nm, respectively. The third mode was believed to be representative of internally mixed Aitken mode particles composed of a soot/ash core with an adsorbed layer of readily volatilisable material. The fourth mode was believed to be composed of chemically aged, secondary particles. The larger nucleation and Aitken modes accounted for between 80-90% of the measured N(TOTAL), and the particles in these modes were believed to be derived from SI and diesel engine emissions. The overall size distribution, particularly in modes II-IV, was observed to be strongly related to the number of primary particle emissions, with larger count median diameters observed under conditions where low numbers of primary soot based particles were present. PMID- 17133278 TI - Rapid analysis of tile industry gaseous emissions by ion mobility spectrometry and comparison with solid phase micro-extraction/gas chromatography/mass spectrometry. AB - The present paper reports on a rapid method for the analysis of gaseous emissions from ceramic industry, based on ion mobility spectrometry (IMS) as a means for on site monitoring of volatile organic compounds (VOCs) produced during tile baking. IMS was calibrated with a set of reference compounds (i.e. ethyl acetate, ethanol, ethylene glycol, diethylene glycol, acetaldehyde, formaldehyde, 2-methyl 1,3-dioxolane, 2,2-dimethyl-1,3-dioxolane, 1,3-dioxolane, 1,4-dioxane, benzene, toluene, cyclohexane, acetone, acetic acid) via air-flow permeation. The technique was tested on a laboratory-scale kiln and tiles prepared with selected glycol- and resin-based additives. Finally, the analytical method was applied to emissions from two industries in the Modena (Italy) ceramic area. The results of all experimental phases were compared to those obtained by solid phase micro extraction/gas chromatography/mass spectrometry (SPME/GC/MS). IMS showed potential as a real-time monitoring device for quality assessment in ceramic industry emissions. IMS spectra, SPME/GC/MS data, relationship between additives/baking conditions and produced VOCs and advantages and limitations of both techniques will be discussed. PMID- 17133279 TI - Microcosm studies on the air-soil exchange of hexachlorobenzene and polychlorinated biphenyls. AB - Soils have an important role to play in supplying and receiving contaminants from the atmosphere, and in the global cycling of certain persistent organic pollutants (POPs). A microcosm study was conducted, in which some aspects of air soil exchange likely to be important for the global cycling of POPs were simulated. HCB and PCBs were introduced into a sealed chamber, which contained soils of varying organic matter content, and in which soil mixing was a variable. Uptake of these compounds on to the soils was monitored over time. Subsequently, the primary chemical source was removed and further changes in the residues retained on the soils were monitored. The rates of uptake onto the different soil treatments are compared and the influence of the soil organic matter content and mixing evaluated. The implications for the global cycling of POPs are discussed. PMID- 17133280 TI - Real-time PCR detection of toxigenic Fusarium in airborne and settled grain dust and associations with trichothecene mycotoxins. AB - Inhalation of immunomodulating mycotoxins produced by Fusarium spp. that are commonly found in grain dust may imply health risks for grain farmers. Airborne Fusarium and mycotoxin exposure levels are mainly unknown due to difficulties in identifying Fusarium and mycotoxins in personal aerosol samples. We used a novel real-time PCR method to quantify the fungal trichodiene synthase gene (tri5) and DNA specific to F. langsethiae and F. avenaceum in airborne and settled grain dust, determined the personal inhalant exposure level to toxigenic Fusarium during various activities, and evaluated whether quantitative measurements of Fusarium-DNA could predict trichothecene levels in grain dust. Airborne Fusarium DNA was detected in personal samples even from short tasks (10-60 min). The median Fusarium-DNA level was significantly higher in settled than in airborne grain dust (p < 0.001), and only the F. langsethiae-DNA levels correlated significantly in settled and airborne dust (r(s) = 0.20, p = 0.003). Both F. langsethiae-DNA and tri5-DNA were associated with HT-2 and T-2 toxins (r(s) = 0.24-0.71, p < 0.05 to p < 00.01) in settled dust, and could thus be suitable as indicators for HT-2 and T-2. The median personal inhalant exposure to specific toxigenic Fusarium spp. was less than 1 genome m(-3), but the exposure ranged from 0-10(5) genomes m(-3). This study is the first to apply real-time PCR on personal samples of inhalable grain dust for the quantification of tri5 and species-specific Fusarium-DNA, which may have potential for risk assessments of inhaled trichothecenes. PMID- 17133281 TI - Measurements of dissolved methylmercury in natural waters using diffusive gradients in thin film (DGT). AB - A diffusive gradient in thin films (DGT) technique for measuring methylmercury (MeHg) concentrations in natural waters was developed using 3-mercaptopropyl functionalized silica gel to preconcentrate the methylmercury. The new resin was characterized and calibrated. Methylmercury is efficiently accumulated at a pH range of 3-9. Basic performance tests of the new DGT device confirmed the applicability of Fick's first law for such DGT measurements. The diffusion coefficient of methylmercury in polyacrylamide gel was 5 x 10(-6) cm(2) s(-1). Methylmercury concentrations determined by DGT deployed for different time periods in the field are statistically not different from results obtained through direct measurements. The DGT technique represents therefore an alternative in situ sampling method for methylmercury. The detection limit of the overall method is 1 pg of MeHg, which correspond to approximately 30 pg L(-1) of MeHg in a water sample, when deploying a typical DGT device for 24 hours. Lower MeHg concentrations are measurable using longer deployment times or thinner diffusive gel layers. PMID- 17133282 TI - Development of a simple extraction cell with bi-directional continuous flow coupled on-line to ICP-MS for assessment of elemental associations in solid samples. AB - A continuous-flow system comprising a novel, custom-built extraction module and hyphenated with inductively coupled plasma-mass spectrometric (ICP-MS) detection is proposed for assessing metal mobilities and geochemical associations in soil compartments as based on using the three step BCR (now the Measurements and Testing Programme of the European Commission) sequential extraction scheme. Employing a peristaltic pump as liquid driver, alternate directional flows of the extractants are used to overcome compression of the solid particles within the extraction unit to ensure a steady partitioning flow rate and thus to maintain constant operationally defined extraction conditions. The proposed flow set-up is proven to allow for trouble-free handling of soil samples up to 1 g and flow rates < or =10 mL min(-1). The miniaturized extraction system was coupled to ICP MS through a flow injection interface in order to discretely introduce appropriate extract volumes to the detector at a given time and with a given dilution factor. The proposed hyphenated method demonstrates excellent performance for on-line monitoring of major and trace elements (Ca, Mn, Fe, Ni, Pb, Zn and Cd) released when applying the various extracting reagents as addressed in the BCR scheme, that is, 0.11 M CH(3)COOH, 0.1 NH(2)OH.HCl and 30% H(2)O(2), even when a well recognized matrix-sensitive detector, such as ICP-MS, is used. As a result of the enhanced temporal resolution of the ongoing extraction, insights into the breaking down of phases and into the kinetics of the metal release are obtained. With the simultaneous multielement detection capability of ICP-MS, the dynamic fractionation system presents itself as an efficient front-end for evaluation of actual elemental association by interelement comparison of metals leached concurrently during the extraction time. Thus, the intimate elemental association between Cd and Zn in contaminated soils could be assessed. PMID- 17133283 TI - [Infectious mononucleosis.]. AB - OBJECTIVE: To present updated aspects of the Infectious Mononucleosis caused by the Epstein-Barr Virus. MATERIALS AND METHODS: Research of bibliographic references through the Medline and direct research of selected papers. RESULTS: A concise approach to some aspects related to the epidemiology of the virus, namely the two types that are presently known,EBVtypeAandEBVtype B, and some differences that they present. It was possible to establish a relationship between what one sees in the clinical picture and the immunological changes that occur at the same time. The author also describes the physiopathology, clinical features, complications and other syndromes associated to the EBV. As far as the laboratory workup is concerned, it is important to have a complete blood count as the first step, followed by the quantitative exam of the heterophile antibodies and antibodies antiEBV analysis. CONCLUSION: The pathologies related to the EBV are important, and certainly fascinating from the immunological point of view. Among these, the Infectious Mononucleosis caused by the EBV has shown some interesting clinical and laboratorial aspects. PMID- 17133284 TI - [The ti-epiplating system in bone anchored hearing AIDS]. AB - The advent of bone-anchored hearing aids with titanium implants has markedly improved patient management. There are two systems on the market for performing the requisite osseointegration. The aim of our study was to evaluate the Ti Epiplating system, since no data have been published on this in the literature thus far. Between November 2002 and March 2005, we anchored the BAHA snap coupling via the Ti-Epiplating system in 8 patients in a single session. The patients were followed-up at regular intervals to assess clinical and audiometric results and subjective satisfaction. The postoperative hearing threshold with the BAHA was in the former bone conduction range. In the Freiburg speech test, a significant improvement was found for monosyllables (free-field), and the hearing loss for numbers showed a mean reduction from 75 to 30 dB. There has been no implant loss in our collective thus far. Good to very good results were evaluated for the quality of life in the "International Inventory of evaluation of hearing aids" (IIEH). Clinics that use Medicon's osteosynthesis system can safely and reliably achieve inexpensive and audiometrically successful BAHA management by a comparatively simple surgical procedure. PMID- 17133285 TI - A comparison of organ-tissue level body composition between college-age male athletes and nonathletes. AB - The purpose of this study was to compare the characteristics of skeletal muscle (SM) mass and internal organ (liver and kidney) mass in resistance and/or high intensity trained collegiate athletes with nonathletes, and to examine the relationships between fat-free mass (FFM) and its major components of SM, liver mass, and kidney mass. Fifteen athletes and seventeen nonathletes volunteered for the study. FFM was measured by two-compartment densitometry. Contiguous magnetic resonance imaging was used to obtain images from the first cervical vertebrae to the ankle joint (no inter-slice gap) for each subject, and SM, liver and kidney cross-sectional areas and organ-tissue volumes were determined. Organ-tissue volumes (cm (3)) were converted to mass (kg) by multiplying the volumes by the assumed constant density of the tissues. On average, athletes had greater FFM (69.1 kg) than the nonathletes (52.6 kg). SM, liver, and kidney masses in athletes (33.0 kg, 1.84 kg and 0.39 kg, respectively) were higher compared with nonathletes (23.5 kg, 1.39 kg and 0.31 kg, respectively). When the various determinants of FFM were expressed as ratios, it was determined that the ratio of SM mass to FFM was higher in athletes (47.7 %) than nonathletes (44.7 %), and the ratios of liver and kidney to FFM were similar for the two groups (2.6 % and 0.6 %, respectively, for athletes; and 2.7 % and 0.6 %, respectively, for nonathletes). A strong correlation between FFM and SM mass was observed in athletes and nonathletes and the slopes of these regression lines were almost identical and parallel. FFM was also significantly correlated with liver and kidney mass for both athletes and nonathletes. This study suggests that SM, liver, and kidney masses are increased by FFM accumulation in resistance and/or high intensity trained athletes. PMID- 17133286 TI - Effect of individual time to peak power output on the expression of peak power output in the 30-s Wingate Anaerobic Test. AB - The objective of the present investigation was to examine a proposal which stated that individual power values should be aligned according to peak power output (PPO) before calculating the mean value of PPO. This procedure removes the variation in time it takes for individuals to reach PPO. Participants were forty one University Rugby Union Football players of mean age 21.7 +/- 2.6 years, height 181.4 +/- 6.9 cm and body mass 88.9 +/- 12.7 kg. Data were collected using a friction-belt cycle ergometer (Monark 864, Varberg, Sweden). A significantly larger mean value for PPO was found when results were calculated from time aligned rather than cross-sectional data (1154 +/- 246 vs. 1121 +/- 254 W, p < 0.0001); the mean difference was approximately 3 %. Additionally, the average profile of the power output curve was more reflective of individual power curves. The negative correlation between PPO and the time taken to reach PPO was - 0.32 (p < 0.05), confirming the view that the earlier the time taken to reach PPO the larger the PPO. It was concluded that the mean value of PPO and the corresponding profile for power output curves are best represented by the analysis of time aligned rather than cross-sectional data. PMID- 17133287 TI - Acceptability of power variation during a simulated hilly time trial. AB - We investigated the acceptability of power variation during a cycling time trial (TT) with simulated uphill and downhill sections. Seven cyclists first completed an 800-kJ self-paced TT on a simulated flat course. An 800-kJ TT course with four sections of uphill/downhill was then modeled. Each section involved 100 kJ of cycling up a simulated gradient of 5 % followed by 100 kJ of riding down a simulated gradient of - 5 %. Participants were required to complete this simulated course using two pacing strategies; (i) at a constant power equivalent to the mean power achieved during the initial TT, and (ii) increasing power by 5 % of mean power when traveling uphill (mean duration of each climb = 714 s) and decreasing power in the downhill sections (mean duration of each descent = 190 s), so that overall mean power was equivalent to that in (i). All participants maintained this variable power strategy during the first half of the TT, but two riders could not adhere to the power variations during the final 400 kJ. Nevertheless, mean +/- SD finish time for the variable power trial (3670 +/- 589 s) was significantly faster than that for the constant power TT (3758 +/- 645 s), the 95 % confidence interval for the percentage improvement being 0.4 to 4.3 %. Heart rate and lactate responses were highest in the initial self-paced TT and did not differ between the subsequent constant and variable power trials. Ratings of perceived exertion were also similar between trials. In our externally-valid TT, we found that some cyclists cannot fully adhere to a pacing strategy involving an approximate +/- 5 % variation in mean power in parallel with gradient variation. Nevertheless, an important time saving can still result even if a variable pacing strategy is only partially adopted during a hilly time trial, so that no additional physiological strain is incurred. PMID- 17133288 TI - Adiponectin oligomers in human serum during acute and chronic exercise: relation to lipid metabolism and insulin sensitivity. AB - Beneficial effects of physical exercise include improved insulin sensitivity, which may be affected by a modulated release of adiponectin, which is exclusively synthesized in white adipose tissue and mediates insulin sensitivity. Adiponectin circulates in three different oligomers, which also have a distinct biological function. We therefore aimed to investigate the distribution of adiponectin oligomers in human serum in relation to physical activity. Thirty-eight lean and healthy individuals were investigated. Seven healthy women and 8 healthy men volunteered to investigate the effect of chronic exercise, at 3 different time points with different training intensities. These individuals were all highly trained and were compared to a control group with low physical activity (n = 15). For studying acute exercise effects, 8 healthy men participated in a bicycle test. Adiponectin was determined by ELISA, oligomers were detected by non denaturating western blot. Total adiponectin and oligomers were unchanged by acute exercise. LDL cholesterol was significantly lower in the chronic exercise group (p = 0.03). Total adiponectin levels and oligomers were not different between these two groups and were unaltered by different training intensities. However, total adiponectin and specifically HMW oligomers correlated with HDL cholesterol (r = 0.459; p = 0.009). We conclude that acute and chronic exercise does not directly affect circulating adiponectin or oligomer distribution in lean and healthy individuals. Whether such regulation is relevant in individuals with a metabolic disorder remains to be determined. However, our data suggest that adiponectin oligomers have distinct physiological functions IN VIVO, and specifically HMW adiponectin is closely correlated with HDL cholesterol. PMID- 17133291 TI - [Renal transplant: color duplex ultrasound and contrast-enhanced ultrasound in the evaluation of the early postoperative phase and surgical complications]. AB - Contrast ultrasound is a promising and straightforward method that is superior to established sonographic techniques such as conventional B-mode scanning which is used for volume measurement and hematoma demonstration. Color Doppler is important for the evaluation of rejection, the detection of perfusion defects, and complete vascularization in the diagnostic evaluation of kidney grafts. Moreover, contrast US has the potential for tumor characterization in transplanted kidneys. A single examination by contrast ultrasound can answer a variety of questions with respect to the early postoperative phase and chronic damage. New applications of contrast US will arise from the further technical development of ultrasound equipment. The rapid technical advances seen in recent years have been followed by the introduction of new software tools for the analysis of raw datasets or the improved visualization of microbubbles at very low energy. Initial studies show that efficient and early diagnosis of rejection is possible. Surgical complications like perfusion defects or hematoma can also be identified. PMID- 17133292 TI - [The untellable patient's history]. AB - An essay with personal touch on the interaction between patients and professionals seen from the perspective of an anonymous writer that is presumably preoccupied with midlife reflections. PMID- 17133293 TI - [Chronic disease management]. AB - Chronic disease management by primary care physician is one of the outstanding challenges in family medicine. Regular checking of possible interactions between different diagnoses and treatments is of great opportunity. Optimal communication and cooperation between doctors in hospital and primary care is a continuous commitment. PMID- 17133294 TI - [More haste less speed]. AB - A 36 year old Portuguese man was admitted to our hospital with diffuse malaise, 25 kg weight loss in the past 6 months and night sweats. The diagnostic workup revealed hyperthyroidism due to graves disease and a mass lesion in the upper anterior mediastinum. Because of the uncertain dignity of the tumor a thoracoscopic thymectomy was performed. Histologically we found a thymic hyperplasia without any signs of malignancy. If systematically searched, thymic hyperplasia is often found with graves disease, however, this association is not widely appreciated and is not mentioned in general text books. The pathophysiology of thymic hyperplasia in graves disease is not fully understood. Immunological mechanisms seem to play an important role, since TSH receptors have been found on human thymic epithelial cells. Thyreostatic therapy leeds to spontaneous shrinkage of the thymic gland. Awareness of thymic hyperplasia in the differential diagnosis of an anterior mediastinal mass lesion is essential, since it prevents further invasive diagnostic procedures that might cause unnecessary morbidity. PMID- 17133295 TI - [Breaking bad news]. AB - Breaking bad news is a difficult task of the physician. With three short records of cancer patients frequent difficulties and errors in the disclosure of diagnosis and prognosis are highlighted. In addition to its medical definition "cancer" has also a metaphoric meaning, which may substantially compromise the communication between patient and physician. Therefore the term cancer has to be explicitly used in talking to the patient and its current meaning for the patient has to be clarified. In estimating the prognosis with the patient precaution is essential since wrong predictions may geopardize the confidence of the patient in this physician substantially and even compromise further treatment. Breaking bad news is not a one way communication. Patient reactions have to be acknowledged, understood and catched up in a supportive way. PMID- 17133296 TI - [From starving to life-threatening nutrition--the history of an intestinal pseudoobstruction]. AB - It is the story of a 70-year-old lady, who suffered from chronic intestinal pseudoobstruction since her adolescence. In the early 90ies progressive cachexia developed. In 1994 parenteral nutrition was begun via a port-a-cath-system with good success in the first years. Later, various complications occurred: thrombotic events, several catheter-related infections with various bacterial strains, an endocarditis of the aortic valve, which was replaced by a bioprosthesis, and finally a relapsing endocarditis of this artificial valve with a life-threatening paravalvular abscess and regurgitation. She also survived this second heart surgery and is currently under parenteral nutrition again, with a more than uncertain future. PMID- 17133297 TI - [Maxillary sinus infection with emphysema of the head as a "red herring" in steroid associated colon perforation]. AB - Steroid therapy increases the risk of bowel perforation. Bowel perforation may occur at any time of steroid therapy, but the first weeks appear to hold the greatest potential for perforation. However, clinical findings after perforation may be misleading under steroids, and peritonitis may be absent. It is known that bowel perforation can lead to subcutaneous emphysema at various sites. Thus, in any patient with emphysema, bowel perforation must be included in the differential diagnosis, especially in patients receiving steroids. Missing knowledge of this entity may lead to marked delay between onset of initial signs and diagnosis, and hence worsen the survival rate. In this report we present a case of chronic steroid use, where asymptomatic sigma perforation led to a generalized emphysema, which was initially attributed to a maxillary sinus infection due to Aspergillus and anaerobic bacteria. PMID- 17133298 TI - [Calcific uremic arteriolopathy]. AB - The history of a patient with calcific uremic arteriolopathy is discussed. The hypothetical pathogenesis and the therapeutic approach is revised. PMID- 17133299 TI - [Somnolence of unknown origin--illness or drug problem?]. AB - A 64 year old female patient with previously known temporal lobe epilepsy (TLE) was admitted to the emergency department (ED) for further investigation of somnolence of unknown origin. Differential diagnosis remained puzzling after exclusion of an intracranial process and status epilepticus. Quantitative and qualitative fluctuations of consciousness are well known to take place in TLE as well as that these patients often are receiving several drugs for their illness. Drug history and measuring serum drug levels are helpful. Adverse Drug Events (ADE) are not rare in EDs: approximately 5% of ED patients are suffering an ADE and about 3% of patients admitted to hospitals are admitted because of an ADE. It is important to realize that three out of four ADEs are estimated to be preventable. PMID- 17133300 TI - [Individualising the evidence--single case randomized trial]. AB - A 64-year old patient with well controlled essential hypertension reported convincingly that oral yohimbine had beneficial acute effects in treating his erectile dysfunction. However, when asking for a prescription of the compound his physician was not very responsive because acute effects of this treatment are uncommon and a2-adrenoceptor antagonism was known to increase blood pressure. They decided to convince each other and to objectively document yohimbine's efficacy or failure using a single case randomized trial design, i.e. an experimental approach to perform a controlled study in a single individual and to collect data in a double-blind fashion. The results of 10 comparisons between placebo and oral yohimbine revealed no difference and prevented exposure of the patient with a potentially harmful drug combination. PMID- 17133301 TI - [In Medea's gardens--two tales of colchicine]. AB - Colchicine is a highly active alkaloid used in the treatment of acute inflammatory syndromes such as Mediterranean fever, M. Behcet or gouty arthritis. The two cases we present here illustrate exemplarily the pros and contras of colchicine therapy. In the first case, colchicine was successfully given for recurrent febrile attacks due to acute rheumatic fever. The second patient unfortunately had a fatal colchicine intoxication. The pharmacology of colchicine, the clinical features associated with overdose and the options for treatment are discussed. Colchicine should not be given in combination with macrolides, especially in patients with renal insufficiency. PMID- 17133302 TI - [Plaque epidemy in the valley of Grindelwald 1669]. AB - Plague epidemy in 1669 in the valley of Grindelwald (Bernese Oberland/ Switzerland) has been described with acrimony by the priest of the village. Despite of success of modern medicine, numerous problems stay practically unchanged for centuries and history will be an important teacher even in the future. PMID- 17133304 TI - [Pain prevalence and patient preferences concerning pain management in the emergency department]. AB - Pain is one of the most common problems for patients who present to the Emergency Department (ED), thus a timely and effective pain management intervention is essential for quality patient care. A comprehensive literature review was undertaken in an effort to document the prevalence of pain and to increase knowledge about patients' preferences regarding pain management in emergency situations. Results indicated that the prevalence of pain is high and pain management, including treatment, is often unsatisfactory in the ED. Common procedures, such as offering pain medication after medical consultation or when diagnostic intervention is completed, were shown not be effective in meeting patient needs. Patients prefer that their pain is treated upon admission to the ED; their need for effective treatment increases with pain intensity. Reasons that patients may refuse pain medication include fear of addiction and side effects. While most patients do not expect being to be pain-free upon discharge, most do expect more effective pain relief than they are currently receiving. Relationships between sociodemographic factors and patient preferences could not clearly be elicited from the literature. A weakness is that the reviewed studies were descriptive and published primarily in the United States in the last five years. There is a need for further research in this area, particularly studies that investigate patients' preferences regarding pain management in European EDs. PMID- 17133305 TI - [Identification of pain in the case of patients under sedation and artificial ventilation: A systematic literature review]. AB - In intensive care units (ICU), patients in life-threatening situations are hospitalized. While the health status of these patients is often severely compromised, therapeutic interventions include endo-tracheal intubation with artificial ventilation, sedation, and analgesia which restrict their ability to communicate. These patients become additional vulnerable since their pains may not been adequately identified. Uncontrolled pain in sedated and artificial ventilated patients lead to increased number of respiration days and increased length of stay in hospital, which, may result in increased morbidity. This situation appears precarious since pain assessment of these patients in daily clinical practice seems often based on subjective criteria by the health care staff in charge. The purpose of this study was to describe pain-assessment instruments for sedated and artificial ventilated ICU-patients in view of validity, reliability and clinical application. With a systematic literature review, a total of 61 articles were identified in the databases of PUB-MED (National Library of Medicine) and CINAHL (Cumulative Index to Nursing and Allied Health Literature ) and additional bibliographic sources. Indicators for pain in sedated and artificial ventilated ICU-patients were physiological and behavior related, without biochemical parameters. In most of the instruments, the validity was tested only superficially and no detailed reports regarding the application in everyday practice were given. However, the studied instruments includes algorithm as a significant component which enable decision making of the health care providers in order to execute stepwise pain management interventions. In conclusion, a need for further research to disclose pain indicators in sedated and artificial ventilated ICU-patients, continuous development of instruments and their validation in clinical practice is obvious. PMID- 17133306 TI - [The meaning of caring from the viewpoint of patients with wounds due to peripheral vascular disease]. AB - This qualitative study describes the meaning of Caring from the viewpoint of the patient with chronic wounds due to peripheral vascular disease (PVD). Because the meaning of Caring can not be understood independent from the life context of the persons involved, the following questions have been formulated: 1) What influence does living with chronic wounds have on the everyday life of the patient? 2) What is the meaning of Caring for patients with chronic wounds? To answer these questions, qualitative research methods according to Mayring were chosen. A convenience sample of twelve patients, seven women and five men, with ages ranging from 69 to 86 years (median 77 years), were interviewed from April to November 2002. Data were analyzed according to Mayring (2000) through content analysis. The results show that the everyday life of the patient is strongly influenced by the effects of having chronic wounds. Patients view their lives with chronic wounds, the effects on their day to day life and the support from health professionals and family caregivers as a whole. Patients differentiate between Caring from Health professionals and Caring from family caregivers. Caring from health professionals is described primarily in the context of treatments and support situations. Caring from family caregivers is experienced as a part of the everyday life. Patients assume that healthcare professionals have the needed expertise, knowledge and skills. They hope, however, to be valued and respected and describe these characteristics as "kind and nice". When patients experience these attitudes from the nurses they feel they are taken seriously and are cared for. This is a requirement to enable or empower patients to understand and implement their treatment regimen. Caring must always be an element of nursing care and determines the standard of the quality of the patient nurse relationship. PMID- 17133307 TI - [How does executive staff of home care nursing services estimate the situation of palliative care?]. AB - Experts criticise heavily the current situation of palliative care in the home care setting in Germany. They claim that the education in palliative care of all involved professional groups should be improved and specialized services should be adjusted. Only few is known in this context about the perspectives and attitudes of nursing staff, who is significantly involved in palliative care. Therefore, interviews with executive stuff of home care nursing services in the Federal State of Brandenburg were performed in the context of an evaluative opinion survey. 88 of 100 planned interviews took place. The nursing services estimated the current situation predominantly good. This does not support the experts' opinions found in the literature. Nevertheless, the respondents saw serious needs for improvements, in particular concerning psycho-social issues. Although palliative patients numerically seem to be in the background of every day work life, great interest in further qualification and new concepts of care was shown. So far, only few home care nursing services employ palliative care trained staff. PMID- 17133308 TI - ["Getting through"--a difficult nursing concept]. AB - This analysis of the research literature related to the nursing concept "enduring" was carried out on the background of the demand for evidence based nursing. The aim was to test with an exemplary concept to what extent this request is feasible and meaningful, if related to a concept describing patients' subjective experience. The state of the art of research literature on enduring contains a variety of problematic aspects: conceptual, theoretical, linguistic as well as problems originating in nursing's moral tradition. These have important implications for the application of the concept in nursing practice. PMID- 17133309 TI - [Visit to the British Center of Nursing History]. PMID- 17133312 TI - Changes in growth hormone (GH) messenger RNA (GH mRNA) expression in the rat anterior pituitary after single interferon (IFN) alpha administration. AB - INTRODUCTION: Interferon alpha (IFN-alpha) is a cytokine with pleiotropic effects which, via different pathways, influences the secretion of certain cytokines and hormones. Growth hormone (GH) secreted from the pituitary has physiological effects on various target tissues. The question is how IFN-alpha administered in various types of disease influences GH secretion. This study investigated the acute effect of IFN-alpha on GH mRNA expression in the rat anterior pituitary. OBJECTIVE: The aim of the study was to measure the cellular expression of GH mRNA by in situ hybridisation in the anterior pituitary after a single administration of IFN-alpha. MATERIAL AND METHODS: Rats were administered an intraperitoneal injection of IFN-alpha or saline. The rat pituitaries were taken 2 and 4 hours after IFN/saline administration and kept frozen until in situ hybridisation histochemistry. A 31-base(35)S-labelled oligonucleotide probe complementary to part of the exonic mRNA sequence coding for GH mRNA was used. All control and experimental sections were hybridised in the same hybridisation reaction. RESULTS: Acute administration of interferon alpha increased GH mRNA expression in the anterior pituitary in the 4-hour group in comparison with the control group, and there was no difference between the control group and the 2-hour rats. CONCLUSION: A single IFN-alpha administration was found to exert an influence on anterior pituitary GH mRNA expression. These observations may pave the way for presenting a possible new action of IFN-alpha. PMID- 17133313 TI - The effect of weight loss on serum concentrations of nitric oxide, TNF-alpha and soluble TNF-alpha receptors. AB - INTRODUCTION: The aims of the present study were to evaluate the effect of weight loss treatment on serum concentrations of NO and TNF-alpha and to examine whether there is an association between TNF-system activity and serum concentrations of NO after weight loss. MATERIAL AND METHODS: The study group involved 43 obese women (aged 41.8 +/- 11.9 years, weight 95.2 +/- 15.0 kg, BMI 36.5 +/- 4.6 kg/m(2)). The women were subjected to three-month complex weight-loss treatment. Patients were advised to keep to a 1000-1200 kcal diet and to exercise regularly. Pharmacological treatment was not administered. Serum concentrations of nitric oxide metabolites, TNF-alpha and its soluble receptors (sTNFR1, sTNFR2) were measured by ELISA kits; insulin was measured by RIA and glucose, cholesterol, HDL cholesterol and triglicerydes by an enzymatic procedure before and after weight loss. Body composition was determined by impedance analysis using Bodystat. RESULTS: The mean weight loss during treatment was 8.3 +/- 4.3 kg. The serum concentrations of TNF-alpha decreased significantly (p < 0.000) and both receptors sTNFR1 and sTNFR2 increased significantly (p < 0.000) after weight loss. No significant changes in serum concentrations of NO were observed after weight loss. A multiple regression analysis was performed using DeltaTNF-alpha, DeltasTNFR1, DeltaTNFR2 and DeltaNO as dependent variables. A significant correlation was observed between DNO and initial plasma concentrations of TNF alpha, sTNFR1 and sTNFR2. CONCLUSIONS: This study demonstrates a decrease in serum TNF-alpha concentration as well as an increase in plasma concentration of both TNF receptors but does not show any change in serum concentrations of NO after weight-loss treatment in obese women. It seems that changes in TNF-system activity may be a counter-regulating mechanism, which inhibits further body mass loss. We did not observe any association between changes in TNF-system activity and serum concentrations of NO after weight loss. PMID- 17133314 TI - Evaluation of bone mineral density on the basis of the results of studies of selected skeleton populations from the microregion of Brzesc Kujawski. AB - INTRODUCTION: Osteoporosis is a systemic disease of the skeletal system characterised by reduced bone strength leading to increased risk of fracture. Bone strength is a combined derivative of bone mineral density (BMD) and of bone tissue quality. Osteoporosis is a serious medical problem because of its complications, most frequently manifesting itself in spine fractures, fractures of distal sections of the forearm and, in later periods of life, hip fractures. Reduced BMD is an independent risk factor of osteoporosis. The goal of the study was an evaluation of bone mineral density of the population inhabiting the micro region of Brzesc Kujawski from the 11(th) century until the beginning of the 19th century. MATERIAL AND METHODS: Human bones obtained from archaeological excavations at four archaeological sites: Kolonia (11th-13th centuries), SBK-4 (12th-16th centuries), Fara (14th-17th centuries) and Swiety Duch (16th-19th centuries) and from the collections of the Katedra of Anthropology of the University of Lodz were subjected to study. Bone mineral densities of the skeleton populations were compared with those of the control group, namely the present living population of the Lodz Province. The incidence of osteoporosis was evaluated by densitometric assessment, which was performed by dual energy X-ray absorptiometry (DXA) on a DPX device (LUNAR, USA). RESULTS: The densitometric measurements performed enabled differences to be identified in the mineral density of the osseous tissue. The skeletal groups studied were characterised by a significantly higher mean BMD than the contemporary living population. Changes in BMD indicative of osteopenia prevailed in women. CONCLUSIONS: On the basis of the results obtained it cannot definitively be stated that the skeletal groups studied, despite their significantly higher BMD, were affected by a lower risk of osteoporosis and its complications. A continuation of studies on the mineral content of bones and on the relationship between the mineral content and bone mineral density is required. PMID- 17133315 TI - [Hypoparathyroidism after surgery on thyroid cancer: is there a delayed chance for recovery after a prolonged period of substitutive therapy?]. AB - INTRODUCTION: Transient and persistent hypoparathyroidism (HPT) belong to the well known complications of total thyroidectomy performed because of thyroid carcinoma. The true frequency of persistent hypoparathyroidism is often higher than estimated in the reports published by the specialized centers with low rate of complications. THE AIM OF THE STUDY: Investigation whether the repeated check up, performed over 2 years post thyroidectomy, reveals some cases of recovery in patients diagnosed with persistent HPT post thyroid cancer surgery. MATERIAL AND METHODS: In total, 115 patients were included into the study, all of them treated with vitamin D derivatives and calcium supplementation. In 17 of them a diagnosis of transient hypoparathyroidism was made on the basis of evaluation performed 6 months after surgery, the remaining 98 were diagnosed with persistent HPT. Parathyroid (PTH) function was reevaluated after withdrawal of active vitamin D derivatives for 10 days and of calcium carbonate for two days during the hospital stay in patients admitted for radioiodine scan, thus after thyroxine withdrawal. The control group consisted of 123 DTC (differentiated thyroid carcinoma) patients without parathyroid dysfunction. On the basis of intact PTH serum level and calcium and phosphorus estimations HPT was unequivocally confirmed in 49 patients (50%). The remaining 49 patients exhibited normal PTH level and in 43 (86%) of them Ca(2+) level was also within normal range, thus delayed, recovery from HPT was stated. RESULTS: Our results indicate that reevaluation of hypoparathyroidism post total thyreoidectomy is necessary, as delayed recover of parathyroid dysfunction is a frequent phenomenon. We also propose criteria of reevaluation of HTP in patients on chronic substitutive therapy. PMID- 17133316 TI - [Transferase S-glutathione class pi gene (GSTP1) polymorphism in thyroid cancer patients]. AB - INTRODUCTION: One of the potential genes which can increase the risk of cancer is GSTP1 gene. It encodes enzyme called glutathione S-transferase pi class, which is involved in the detoxification of a variety of potential carcinogenic compounds. Polymorphism in this gene can cause the amino acid substitution. This substitution, close to the substrate binding site, changes the enzymatic activity for particular substrates and subsequently increases the risk of carcinogenesis. The aim of this study was to evaluate the function of GSTP1 polymorphism in thyroid cancer and possible association between GSTP1 polymorphism and age at diagnosis. MATERIAL AND METHODS: 103 patients with differentiated thyroid cancer (DTC) and 53 individuals from control group were examined using PCR-RFLP. RESULTS: Statistically insignificant association of studied polymorphisms with thyroid cancer was observed. Comparison of allele frequency between cases and control groups revealed the presence of risk alleles. For the first polymorphism Ile OR = 1.257; 95% CI [0.792-1.997] (p = 0.332), and for the second one Val OR = 1.283; 95% CI [0.6260-2.631] (p = 0.495). The presence of Val/Val (c.313A>G) led to a significant earlier age of onset as compared with other genotypes (p < 0.05). Mean age at diagnosis for Val/Val genotype was 41.1 +/- 15.2, and for Ile/Val + Ile/Ile reached 48.9 +/- 13.2. There was no association between age and genotype for c.341C>T polymorphism. CONCLUSIONS: Statistically insignificant association of GSTP1 gene polymorphism with thyroid cancer was observed in studied group of patients. The Val/Val genotype for c.313A>G polymorphism led to earlier age of tumour diagnosis as compared with other genotypes. PMID- 17133317 TI - [Atypical forms of hyperthyroidism]. AB - Except from well-known the most frequent reasons of the hyperthyroidism such as the Graves-Basedow disease, multinodular goitre and the autonomous adenoma we should also remember the other rarer illnesses leading to the excess of thyroid hormones in the serum. Authors presented the problem of atypical forms of thyrotoxicosis which run without the overproduction of thyroid hormones and are characterized by low 24 h 131 J uptake by the thyroid gland. So called masks of hyperthyroidism such as apathetic, neuromuscular, gastrointestinal and cardiovascular forms were also discussed in the article. PMID- 17133318 TI - [Menopause - facts and controversies]. AB - Menopausal period induces some hormonal changes in female organism. These alterations are responsible for many illnesses and ailments - e.g. disorders of menstrual cycle, climacteric symptoms, urine incontinence, connective tissue lesions, postmenopausal osteopenia and osteoporosis. All the changes negatively influence quality of life of women. The principal element of menopausal care in this period is hormone therapy (HT), alleviating many of the climacteric aliments. This article is focused also on some controversial topics concerning the HT action on breast, coronary system and brain. PMID- 17133319 TI - [A special case of thyroid associated ophthalmopathy in the course of Graves Basedow disease]. AB - INTRODUCTION: The exact pathogenesis of Graves' ophthalmopathy and the possibility of causal treatment of this disease still remain unclear. Currently no standard treatment guidelines have been accepted. While treatment procedures have been established in specialized centres, management of complicated and long lasting cases is always individual. We present an unusual case of Graves' ophthalmopathy accompanied by other autoimmune diseases. CASE REPORT: Our patient, MB, female, born in 1961, was diagnosed with Graves' disease 13 years ago. Recurrent hyperthyroidism and large goitre qualified her for strumectomy (performed twice) and long-term antithyroid treatment. Four years after her initial diagnosis, relapsing severe (ophthalmopathy index: 9 points, CAS: 7 points) occurred which persisted despite continuous administration of glucocorticoids. Due to imminent blindness, orbital decompression had to be performed, three times since. Concurrent autoimmune diseases: ulcerative colitis and seronegative rheumatoid arthritis were also stated. Two years ago, due to loss of vision acuity, rapid progression of exophthalmos and recurrence of hyperthyroidism, immunosuppressive treatment with azathioprine was undertaken over a period of 12 months. The present condition of the patient is satisfactory. CONCLUSION: Judging from the discussed course of treatment, in rare and difficult cases of proliferative ophthalmopathy, early immunosuppressive treatment other than glucocorticoids, should be considered. PMID- 17133320 TI - [Graves-Basedow disease after allogeneic bone marrow transplantation]. AB - One severe aplastic anaemia case who presented autoimmune thyroid disease after allogeneic bone marrow transplantation (alloBMT) is described. A 19 year old Polish girl developed Graves' hyperthyroidisms 19 months after allogeneic BMT for severe aplastic anaemia (SAA) donated from her brother. Her serum was positive for thyroid stimulating antibody (TSAb) and anti-thyroid peroxidase autoantibodies (aTPO) while her brother remained euthyroid, seronegative for TSAb, and showed no clinical signs of thyroid pathology. The genetic studies of lymphocytes FISH (fluorescence in situ hybridization) and analysis of STR (short tandem repeated) fragments suggested, that lymphocytes responsible for hyperthyroidisms were of donor origin. PMID- 17133321 TI - [Diagnosis of Cushing syndrome - contemporary views]. PMID- 17133322 TI - Abstracts of the 2nd general polish conference on neuroendocrine tumors of gastrointestinal system, 2006. PMID- 17133323 TI - [The prevalence and pharmacological cost of Parkinson's disease in Spain]. AB - INTRODUCTION: Parkinson's disease is the second most frequent neurodegenerative disease. AIM: To analyze the dispensation of antiparkinsonian agents in Spain and to estimate the Parkinson's disease prevalence. MATERIALS AND METHODS: Dispensation of antiparkinsonian agents were studied in Spain during two years (October 1998 to September 2000). Results were expressed in defined daily dosages per 1,000 inhabitants per day (DID). Levodopa's DID was used to estimate the prevalence of the disease. The cost per 1,000 inhabitants per day (CID) and the daily treatment cost was also valued. RESULTS: The most frequently used drugs are levodopa, biperiden and selegiline. The total cost reached values of 116,346,589.30 euros during the study period. The CID was 4,14 euros. It was very high the daily treatment cost of pramipexol and entacapone. The prevalence of Parkinson's disease is considered in 1.7 per 1,000 inhabitants in Spain. There is an important geographical variability; regions as Castilla-Leon, Galicia and La Rioja have a higher prevalence than Andalucia or Murcia. The number of patients in Spain can be considered in 69,571 people. CONCLUSION: There are some differences between the autonomous communities in the antiparkinsonian drugs' utilization. PMID- 17133324 TI - [Neuroepidemiology profile of the central zone of the department of Caldas (Colombia), years 2004-2005]. AB - INTRODUCTION: The neurological disease (ND) is a challenge for the medical community, cause of a high degree of morbility, mortality, deterioration in the quality of life of the patient and its family. AIM. To determine the prevalence of ND in the population of the department of Caldas (Colombia): migraine, movement disorders, cerebrovascular diseases, peripheral neuropathy, epilepsy, dementia and insomnia. This last one is innovating aspect in the published Colombian studies has not considered and therefore its prevalence with World Health Organization (WHO) methodology is not known. PATIENTS AND METHODS: This is a population, descriptive study, of cross section. The neuroepidemiological protocol developed by the WHO, modified for Colombia was used. The suspicious cases to suffer ND were evaluated by the neurologist. The information was processed in the statistical package Epi Info 6.04d. RESULTS: Of 787 interviews, 360 were suspicious of ND; additionally 20 non suspicious subjects were evaluated like controls. The global prevalence of ND, excluding insomnia was 13.4%, while including insomnia was 60.4%. The prevalence diseases in sequence (by 1,000 inhabitants) were: insomnia 468 (CI 95%: 433.4-504.4), migraine 278 (CI 95%: 246.3-310.2), dementia (in older 50 years) 51 (CI 95%: 15.8-87), peripheral neuropathy 48 (CI 95%: 32.7-63.4), movement disorders 25 (CI 95%: 13.8-37.0), epilepsy 24 (CI 95%: 12.8-35.5) and cerebrovascular disease 10 (CI 95%: 2.5 17.8). CONCLUSIONS: These results show high prevalence of migraine in our community. Also, the peripheral neuropathy, movement disorders and epilepsy were more frequent than in previous investigations. Dementia and stroke presented prevalence similar to the reported one. The insomnia showed very high prevalence, but by the non possibility of comparison with other local studies, it is need conduced others studies to confirm these findings. PMID- 17133325 TI - [Communication based on the P300 component of event-related potentials: a proposal for a matrix with images]. AB - INTRODUCTION: For more than two decades, several research groups have tried to build a device called "brain computer interface" (BCI) to make it available for people having several disabilities such as the locked in syndrome through the use of the recording of electroencephalography activity while the patients are being visually stimulated. AIM. To obtain a P300 component elicited by intensifications of images arranged in a matrix in an oddball paradigm in two selection modes: assigned and free. SUBJECTS AND METHODS: A 5 x 5 matrix for communication purposes was used to visually stimulate 12 volunteers while their event related potentials were recorded in three leads (Fz, Cz and Pz). Off-line analyses were performed to obtain the P300 component which was elicited by targets images intensified randomly by rows or columns. RESULTS: In both modalities assigned and free, all volunteers generated a reliable P300 component. Confirmation of the selected images was made through a comparison of the P300 when each target, row and column matched. In the free selection mode, higher amplitude and a broader activation including frontal leads was observed. No significant differences in the P300 latency were found. CONCLUSION: In all volunteers the elicited P300 allows the identification of the selected images in the 5 x 5 matrix. In the present study the use of drawings representing ideas instead of letters might increase the communication rate in a P300-based BCI. PMID- 17133326 TI - [Multiple mononeuropathy associated to treatment with pravastatin]. AB - INTRODUCTION: The first reports of neuropathy due to treatment with statins appeared in 1994, although it is an infrequent complication. It usually consists of an axonal polyneuropathy, which is predominantly sensory, distal and symmetric, and may be subacute or chronic. We present here the second case reported in the literature of multiple mononeuropathy associated to the use of statins. CASE REPORT: A 51-year-old female patient who, after beginning therapy with pravastatin, presented with progressive, asymmetrically distributed, distal paresthesias in the limbs and an unstable gait. An electromyographic study was compatible with multiple mononeuritis. Results of complementary tests that were carried out to preclude other causes of multiple mononeuropathy were normal. The patient's condition improved on withdrawing treatment with the drug and it became worse again when therapy was restarted. When pravastatin therapy was stopped for good, the patient's condition progressively improved until she was practically free of symptoms. CONCLUSIONS: The relationship between treatment with statins and the appearance of polyneuropathy has been proved in different epidemiological case-control studies. It does not only appear as the classical distal symmetrical polyneuropathy, but has also been reported as taking on atypical clinical forms including a few cases, like ours, of multiple mononeuropathy. The risk of developing this complication is low and is offset by the cardiovascular benefits offered by statins, although it may become more common in the future due to the increasing rate of use of these agents. It is important to bear this cause of neuropathy in mind, given the fact that it is potentially reversible. PMID- 17133327 TI - [Induction of pseudoseizures by placing inactive electrodes in the malar regions]. AB - INTRODUCTION: Pseudoseizures are paroxysmal disorders of a psychogenic origin with manifestations that are similar to epileptic seizures. They constitute a pathology that is found relatively often in the clinical practice of neurology. In many cases the diagnostic suspicion of their existence is difficult to confirm. The complementary examination that proves to be most useful in diagnosing them is video electroencephalogram (video-EEG). CASE REPORTS: We report the cases of seven patients who were referred to our service with a clinical suspicion of pseudoseizures. Placing electrodes with an inactive surface coating over the two malar regions during a video-EEG study enabled us to trigger in all patients (by means of suggestion) the appearance of a clinical picture that was similar to those that initially gave rise to the request for an examination. Suggestion was also employed to limit the time they lasted by getting the patient to resolve the episode. In all the cases the trace of the simultaneous electroencephalogram showed no pathological findings. CONCLUSIONS: Placing inactive electrodes on malar regions is a method for inducing pseudoseizures that is harmless, easily applicable, readily available and economical, and which has not been reported in the literature. Video-EEG monitoring time is reduced using this approach. It provides evidence that a specific seizure phenomenon can be induced and resolved by suggestion techniques, which is a fact that is very useful in the diagnosis of pseudoseizures. PMID- 17133328 TI - [From energetics to neurotransmission: adenosine triphosphate and its receptors]. AB - INTRODUCTION AND AIMS: Adenosine triphosphate (ATP) is a substance that has stood out for being an energy donor, but today we know that it also has other functions. By activating a large variety of receptors that are widely distributed throughout the nervous system, ATP plays a role in neurotransmission, neuromodulation, apoptosis, cell proliferation and differentiation, pain, and so forth. AIMS: The aim of this study is to carry out a review of ATP receptors, their contribution to the correct functioning of neuronal circuits, and their involvement in the development of certain diseases. DEVELOPMENT: The issue is approached from different perspectives, such as the structure of ATP, its metabolism through ecto-ATPases and their main metabolite, adenosine, in addition to a detailed description of each of the purinergic receptors whose existence has been internationally accepted. We also examine how ATP and its receptors are involved in the structures of the nervous system, and more especially those that correspond to the sensory systems. CONCLUSIONS: The fact that purinoceptors are so widely distributed makes them worthy of a detailed examination. This new family of receptors promises to be a new diagnostic and therapeutic tool that enables us to approach certain aspects of the neurosciences from a new angle, by taking into account a new protagonist, ATP, in the generation of events that upset health. PMID- 17133329 TI - [Executive dysfunctions in adults with attention deficit hyperactivity disorder]. AB - INTRODUCTION: Several different follow-up studies have shown that attention deficit hyperactivity disorder (ADHD) can persist into adulthood. AIM: To review the findings in adults with ADHD related to alterations in the executive functions. DEVELOPMENT: Research conducted among children with ADHD has revealed the existence of alterations in different tasks that evaluate the executive functions, such as the planning test, sustained attention tasks, cognitive flexibility, verbal fluency and working memory tasks, as well as several inhibition response tasks. In adults with ADHD, despite the lower number of reports in the literature and the methodological shortcomings that exist in some studies, analogous results have also been described with respect to executive functioning, namely, disorders affecting inhibition response, the capacity for planning, difficulties in cognitive flexibility and verbal fluency, and problems with working memory, which include aspects of spatial working memory, logical or visual memory. CONCLUSIONS: The findings we have available at present enable us to confirm the persistence of executive dysfunctions in adult patients with ADHD that are similar to those observed in children with ADHD. PMID- 17133330 TI - [Semantic dementia, a many-worded disorder]. AB - INTRODUCTION: Semantic dementia is a progressive, relatively selective disorder affecting the semantic system with involvement of the verbal and non-verbal functions. The clinical picture is well characterised despite the confusion that may be generated by the different ways of classifying it. AIM. To determine the clinical, neurolinguistic, imaging and pathological features of this progressive language disorder. DEVELOPMENT: Evaluation of language reveals above all the existence of semantic paraphasias, disorders affecting the comprehension of isolated words and surface dyslexia. Flow of speech, complex syntactic comprehension and grammar are preserved. Both episodic and autobiographic memory are close to normality. Both the clinical signs and symptoms and imaging studies agree on the fact that the most heavily affected area is the anteroinferomedial region of the temporal lobe on a bilateral scale but with predominance of the left-hand side. Pathologically, in most cases positive intraneuronal ubiquitin inclusions are observed like those described in motor neuron diseases. CONCLUSIONS: Semantic dementia constitutes a diagnosis challenge, mainly from the neuropsychological point of view. Further advances towards reaching a diagnosis would allow us to determine which area is mainly affected and, in the future, to find an effective treatment for this progressive, degenerative disorder. PMID- 17133331 TI - [The origins of language: an analysis from the aphasia perspective]. AB - INTRODUCTION: Different areas of knowledge have contributed to a better understanding of the origins of human language. AIM. To relate our current knowledge about the origins of language with the language pathology found in the case of brain injuries (aphasia). DEVELOPMENT: There are two fundamental forms of aphasia, which linked to defects in the lexico-semantic and grammatical systems of language (Wernicke-type aphasia and Broca-type aphasia, respectively). From observations made on children's development of language and experiments with primates, it has been shown that language initially appears as a lexico-semantic system. Grammar correlates with the ability to represent actions (verbs) and depends on what is known as Broca's area and its related brain circuits, but it is also related to the ability to quickly carry out the sequencing of the articulatory movements required for speaking (speech praxis). CONCLUSIONS: Language may have appeared as a lexico-semantic system much earlier than language as a syntactic system. The former may have developed around 200,000-300,000 years ago, coinciding with the increase in the temporal lobe, and would have existed in other hominids. Language as a grammatical system appeared perhaps as recently as 50,000 years ago and seems to be exclusive to Homo sapiens. PMID- 17133332 TI - [Diagnosis and treatment in a case of respiratory myoclonus]. PMID- 17133333 TI - [Carbamazepine-associated pseudolymphoma]. PMID- 17133334 TI - [Persistent neutrophilic meningitis]. PMID- 17133335 TI - [Embryopathy due to maternal phenylketonuria: a cause of mental retardation]. PMID- 17133336 TI - Optimization of microfabricated nanoliter-scale solid-phase extraction device for detection of gel-separated proteins in low abundance by matrix-assisted laser desorption/ionization mass spectrometry. AB - A nano-scale solid-phase extraction (SPE) device was developed for the detection of gel-separated proteins in low abundance by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) with a simplified microfabrication technology. By using SU-8 photoresist instead of epoxy glue to connect the microchannel and transfer capillary, polymeric contaminant signals in MS analysis were significantly reduced. Micro SPE columns with different capacities and geometric characteristics were investigated in order to increase the detection sensitivity and decrease spot size for MALDI-TOF MS analysis. It is shown that enhancements in sensitivities for the detection of proteins in low abundance were correlated with the reduction in column capacity and increase in column aspect ratio. Fifty nanoliters of matrix solution were sufficient to elute the sample completely from the optimized micro SPE column with 3.5 nL capacity. The mass spectrum of a 5 fmol in-gel tryptic digest of bovine serum albumin (BSA), processed by the micro SPE column, demonstrated that 29 peptides matched the protein giving a sequence coverage of 51%, which was better than that obtained from analysis of 25 fmol of the same sample prepared by the dried-droplet method. With the micro SPE column treatment of 2 microL of digestion supernatant of a gel spot of the IQGAP1 protein, 15 peptides were detected from the mass spectrum with the highest individual score of 111, while, with a ZipTip procedure, only nine peaks were detected with the highest individual score of 71. Analytical results demonstrated that this approach greatly improved the sequence coverage and identification specificity for the tested protein. It can serve as a very useful tool in proteomics studies, especially for low abundance proteins. PMID- 17133337 TI - Analysis of succinylacetone, as a Girard T derivative, in urine and dried bloodspots by flow injection electrospray ionization tandem mass spectrometry. AB - Flow injection electrospray ionization tandem mass spectrometric methods for succinylacetone (SA) in 250 microL urine, using d5-SA as internal standard, and in 3 mm dried bloodspots, using 13C4-SA as internal standard, are described. Selectivity and sensitivity of analysis is achieved by the use of a mono-Girard T derivative. Measured SA infant urine normal range (n=20) is 0.013-0.27 micromol/mmol creatinine. Measured SA newborn bloodspot normal range (n=152) is 0 0.30 micromol/L. Bloodspots from children with hepatorenal tyrosinemia type 1, and kept at room temperature for up to 7 years, afforded SA concentrations of 0.9 5.7 micromol/L. PMID- 17133338 TI - Restituting intestinal epithelial cells exhibit increased transducibility by adenoviral vectors. AB - BACKGROUND AND AIMS: While mature enterocytes are resistant to transduction by adenovirus type 5 (Ad5) vectors, undifferentiated cells are transduced much more efficiently. Our purpose was to study enterocyte transduction in models of intestinal wound healing. METHODS: Transduction was studied ex vivo using cultures of endoscopic biopsies and in vitro utilizing Caco-2 cells in models of mucosal wound healing. Vectors carried either the LacZ or the luciferase gene. CAR (coxsackievirus and adenovirus receptor) and integrins were studied with transduction inhibition and immunofluorescent staining. RESULTS: Increased transduction efficiency was observed for a subset of enterocytes with a flattened de-differentiated phenotype present at the edge of cultured biopsies. In the in vitro systems, expanding Caco-2 cell monolayers exhibited increased transducibility that was time- and dose-dependent, reaching virtually 100% in cells along the leading edge at high viral load. Bioluminescence activity of transduced expanding monolayers was up to 3-fold greater than that of non expanding monolayers ('fence' system, 48 h, MOI 1000, p < 0.05). Mitomycin C pre treatment did not affect levels of transduction in expanding monolayers. At the highest viral load tested, CAR or integrin blocking prior to virus application resulted in 39.4% and 45.4% reduction in transduction levels (p < 0.05). Immunofluorescence revealed altered expression of CAR on the migrating edge of the Caco-2 cultures and the expression of CAR on the apical membrane of biopsy enterocytes. CONCLUSIONS: Increased CAR and integrin accessibility in migrating enterocytes mediates increased transduction by Ad5 vectors. This subset of enterocytes provides a target for the delivery of genes of interest for both research and gene therapy applications. PMID- 17133339 TI - A branched histidine/lysine peptide, H2K4b, in complex with plasmids encoding antitumor proteins inhibits tumor xenografts. AB - BACKGROUND: In this study we investigated whether a particular branched HK polymer, H2K4b, was an effective in vivo carrier of plasmids expressing the antiangiogenic kringle 1-5 or the tumor suppressor p53. METHODS: H2K4b was synthesized on a solid-phase peptide synthesizer. Distribution, optimization and time course studies were done in tumor-bearing nude mice by systemically administering H2K4b in complex with a luciferase-expressing plasmid. We examined the amount of tumor angiogenesis in C6 with MDA-MB-435 xenografts utilizing the carmine dye. The ability of H2K4b to carry luciferase plasmids to different tissues was compared with several liposomal carriers. Medium from cells transfected with mKr1-5 was tested for its capacity to inhibit angiogenesis with an in vivo Matrigel assay. We then determined if systemically delivered H2K4b in complex with plasmid encoding mKr1-5 inhibited tumor growth; we also compared the antitumor activity of HK polyplexes containing hKr1-5, mKr1-5, and p53 plasmids. RESULTS: H2K4b carried the luciferase-expressing plasmid in order of descending efficacy to these tissues: lung, spleen, tumor, and liver. Compared to DOTAP containing liposomes, H2K4b was a more effective carrier of a luciferase containing plasmid to extrapulmonary tissues. We then determined that mKr1-5 in complex with H2K4b reduced MDA-MB-435 tumor growth by approximately 50% compared to the control group (P < 0.01). Similarly, H2K4b/mKr1-5 polyplexes reduced the growth of C6 xenografts. In MDA-MB-435 xenografts, p53- and Kr1-5-expressing plasmids in complex with H2K4b had comparable antitumor activity. CONCLUSION: H2K4b demonstrates potential as a carrier of plasmids encoding antiangiogenic and/or tumor suppressor proteins in a tumor-bearing mouse model. PMID- 17133340 TI - A new inducible adenoviral expression system that responds to inflammatory stimuli in vivo. AB - BACKGROUND: Gene transfer using inducible promoters, which control expression of transgenic proteins in response to physiological conditions, may have significant advantages. In this study, we tried to achieve an inducible adenoviral expression system for physiologically responsive gene therapy of autoimmune or inflammatory diseases. METHODS: A luciferase reporter vector with a hybrid promoter containing the human IL-1beta enhancer region (-3690 to - 2720) and the human CIITA promoter IV (-399 to + 2) was constructed. A replication-deficient adenovirus was engineered with luciferase controlled by the IL1beta/CIITApIV promoter (Ad IL1beta/CIITApIV-Luc). The reporter vector or adenovirus was transfected to C57Bl/6 myeloid dendritic cells (DCs), RAW264.7, and Hep G2 to study the in vitro characteristics of this hybrid promoter. An inflammation model was prepared by injecting lipopolysaccharide (LPS) into Balb/c mice intraperitoneally (i.p.), and infected with Ad-IL1beta/CIITApIV-Luc or Ad-CMV-Luc to study the in vivo characteristics of the IL1beta/CIITApIV promoter. RESULTS: The IL1beta/CIITApIV hybrid promoter has pronounced promoter activity, broad-range responsiveness to cytokines or LPS, and can be rechallenged after first induction. In the inflammation model, IL1beta/CIITApIV could drive hepatic luciferase expression increasedly rapidly after LPS challenge and in a LPS dose-dependent manner. CONCLUSIONS: Using the IL1beta/CIITApIV hybrid promoter in gene transfer vectors may make it possible to produce transgenic proteins in vivo in direct relationship with the intensity and duration of an individual's status. By providing endogenously controlled production of transgenic proteins, this approach might limit the severity of autoimmune or inflammatory response without interfering with the beneficial components of host defense and immunity. PMID- 17133341 TI - One of three successfully treated CGD patients in a Swiss-German gene therapy trial died due to his underlying disease: A position statement from the European Society of Gene Therapy (ESGT). PMID- 17133344 TI - Multiple binding of methyl-CpG and polycomb proteins in long-term gene silencing events. AB - Epigenetic regulation is involved in the maintenance of long-term silencing phenomena, such as X-inactivation and genomic imprinting in mammals. Gene repression is mediated by several mechanisms, such as histone modifications, DNA methylation, and recruitment of Polycomb proteins. To understand the mechanistic relationships between these mechanisms for stable gene silencing, we analyzed the mechanisms of X- and Y-inactivation of the PAR2 gene SYBL1, previously showed to be regulated by concerted epigenetic mechanisms. Maintenance of stable repression occurs via the recruitment of both MBDPs and PRC2 complexes to SYBL1 promoter. Their binding is equally sensitive to defective DNA methylation seen in cells derived from ICF syndrome patients. Multiple occupancy is a feature shared within long-term repressed genes, such as the X-inactivated PGK1 and the imprinted IGF2. MBD2, MBD3, and MeCP2 occupy SYBL1 promoter simultaneously, as revealed by sequential ChIP. We did not find this co-occurring binding when looked for members of PRC2 complex together with any of the methyl-binding proteins. Furthermore, in co-transfection assays, MECP2 can silence methylated SYBL1 promoter, whereas the mutated protein fails. However, RNA interference of endogenous MECP2 does not induce the expression of the inactive SYBL1 alleles, suggesting that its silencing activity can be replaced by the other methyl binding proteins. Our data suggest that maintenance of long-term silencing involves multiple layers of epigenetic control functionally redundant. PRC2 and MBD proteins could collaborate to different phases of this process, the former possibly recruiting DNMTs to the silenced promoters, the latter dictating the lock of the transcription. PMID- 17133345 TI - Differential signal transduction of alternatively spliced FGFR2 variants expressed in human mammary epithelial cells. AB - Gene amplification and protein overexpression of fibroblast growth factor receptor 2 (FGFR2) characterize the SUM-52 breast cancer cell line developed in our laboratory. SUM-52 cells express nine distinct alternatively spliced isoforms of FGFR2. Among these isoforms are two otherwise identical FGFR2 variants that express either the C1 or C3 carboxyl terminus. FGFR2-C3 variants are not normally expressed by human mammary epithelial (HME) cells, and we have shown that overexpression of FGFR2-C3 in HME cells results in potent transformation. In particular, FGFR2-C3 expression leads to robust levels of constitutively tyrosine phosphorylated FRS2 in the absence of ligand stimulation. In contrast, overexpressed FGFR2-C1 requires constant stimulation with exogenous keratinocyte growth factor (KGF) to mimic the signaling capability of FGFR2-C3. However, activation of FRS2 that results from KGF-stimulated FGFR2-C1 signaling is transient and is associated with a mobility shift of FRS2 not observed when this signaling molecule is activated by the C3 isoform of FGFR2. Mutation of the only tyrosine phosphorylated site present in the C1 terminus and absent from C3, Tyr769, did not yield a receptor that rivaled the potent signaling of FGFR2-C3. We therefore conclude that aberrant expression of alternatively spliced isoforms of FGFR2 with the C3 carboxyl terminus in the SUM-52 breast cancer cells results in sustained activation of signal transduction leading to transformation. PMID- 17133346 TI - Molecular targets for retinal vascular diseases. AB - The elucidation of the molecular pathogenesis of a disease in animal models provides candidate targets for treatment. As specific antagonists for a target are developed and tested in clinical trials, if benefit is achieved, the candidate becomes a validated target. Validated targets stimulate additional research to identify optimal ways of attacking the target and studies in related disease processes to determine if the molecule is also a target in that context. Vascular endothelial growth factor (VEGF) has been identified as a validated target for several retinal vascular diseases. This has led to a flurry of activity resulting in beneficial treatments for patients and intensification of the search for other targets. This review summarizes preclinical and clinical trial results obtained with VEGF antagonists and describes evidence supporting the candidacy of other molecules currently being tested or soon to be tested for target status. PMID- 17133347 TI - Tristetraprolin recruits functional mRNA decay complexes to ARE sequences. AB - AU-rich elements (AREs) in the 3' untranslated region (UTR) of numerous mammalian transcripts function as instability elements that promote rapid mRNA degradation. Tristetraprolin (TTP) is an ARE-binding protein that promotes rapid mRNA decay through mechanisms that are poorly understood. A 31 nucleotide ARE sequences from the TNF-alpha 3' UTR promoted TTP-dependent mRNA decay when it was inserted into the 3' UTR of a beta-globin reporter transcript, indicating that this short sequence was sufficient for TTP function. We used a gel shift assay to identify a TTP-containing complex in cytoplasmic extracts from TTP-transfected HeLa cells that bound specifically to short ARE sequences. This TTP-containing complex also contained the 5'-3' exonuclease Xrn1 and the exosome component PM-scl75 because it was super-shifted with anti-Xrn1 or anti-PMscl75 antibodies. RNA affinity purification verified that these proteins associated specifically with ARE sequences in a TTP-dependent manner. Using a competition binding assay, we found that the TTP-containing complex bound with high affinity to short ARE sequences from GM-CSF, IL-3, TNF-alpha, IL-2, and c-fos, but did not bind to a U-rich sequence from c-myc, a 22 nucleotide poly U sequence or a mutated GM-CSF control sequence. High affinity binding by the TTP-containing complex correlated with TTP dependent deadenylation and decay of capped, polyadenylated transcripts in a cell free mRNA decay assay, suggesting that the TTP-containing complex was functional. These data support a model whereby TTP functions to enhance mRNA decay by recruiting components of the cellular mRNA decay machinery to the transcript. PMID- 17133348 TI - Granulocyte colony-stimulating factor promotes the translocation of protein kinase Ciota in neutrophilic differentiation cells. AB - Previously, we suggested that the phosphatidylinositol 3-kinase (PI3K)-p70 S6 kinase (p70 S6K) pathway plays an important role in granulocyte colony stimulating factor (G-CSF)-dependent enhancement of the neutrophilic differentiation and proliferation of HL-60 cells. While atypical protein kinase C (PKC) has been reported to be a regulator of p70 S6K, abundant expression of PKCiota was observed in myeloid and lymphoid cells. Therefore, we analyzed the participation of PKCiota in G-CSF-dependent proliferation. The maximum stimulation of PKCiota was observed from 15 to 30 min after the addition of G CSF. From 5 to 15 min into this lag time, PKCiota was found to translocate from the nucleus to the membrane. At 30 min it re-translocated to the cytosol. This dynamic translocation of PKCiota was also observed in G-CSF-stimulated myeloperoxidase-positive cells differentiated from cord blood cells. Small interfering RNA for PKCiota inhibited G-CSF-induced proliferation and the promotion of neutrophilic differentiation of HL-60 cells. These data indicate that the G-CSF-induced dynamic translocation and activation processes of PKCiota are important to neutrophilic proliferation. PMID- 17133349 TI - Lamins of ocular lens epithelial cells. AB - Experiments were performed to characterize a prominent nuclear matrix (NM) protein isolated from tissue cultured mouse lens epithelial cells. This NM protein was separated by SDS-PAGE and the stained gel band was analyzed by mass spectroscopy. Blast analysis of the amino acid sequence derived by mass spectroscopy revealed the presence of Lamin C in the NM of the mouse lens epithelial cells. We also examined nuclear proteins of adult and fetal human lenses. Data collected from these experiments showed the presence of Lamin C in both adult and fetal lens cells. However fetal lens cells only show Lamin C dimers, whereas adult human lens contained dimers, monomers and degraded Lamin C. Early and late passaged tissue cultured mouse lens epithelial cells also contained Lamin C in the nucleus with a preponderance of the dimer in the early passaged cells. The biological significance of the presence of dimers in human fetal lens cells and early passaged mouse lens cells is not known. However, it could suggest an enhanced docking capability of Lamin C dimers for other physiologically important nuclear proteins. PMID- 17133350 TI - Contraction-induced interleukin-6 transcription in rat slow-type muscle is partly dependent on calcineurin activation. AB - The present work aimed at determining whether interleukin-6 (IL-6) produced by skeletal muscle during exercise is related, at least partly, to calcineurin activity. Rats were treated with two specific calcineurin inhibitors, cyclosporin A (CsA) and FK506, or vehicle (Vhl); they were then subjected to exhaustive treadmill running. Modulatory Calcineurin-Interacting Protein-1 (MCIP-1) mRNA levels, a reliable indicator of calcineurin activity, and IL-6 mRNA levels were measured by real-time RT-PCR in soleus muscles, and IL-6 protein concentration was measured in the plasma. Because low carbohydrates availability enhances IL-6 transcription through p38 Mitogen Activated Protein Kinase (MAPK) pathway, muscle glycogen content and glycaemia were measured and p38 MAPK phosphorylation was determined in skeletal muscle by western blotting. As expected, exercise induced an increase in IL-6 (P < 0.01) and MCIP-1 mRNA (P < 0.01) in soleus muscle of Vhl rats, and enhanced p38 phosphorylation and plasmatic IL-6 protein (P < 0.05). Calcineurin inhibition did not affect running time, glycemia or soleus glycogen content. CsA administration totally inhibited the exercise-induced increase in MCIP-1 mRNA (P < 0.01), blunted the IL-6 gene transcription related to muscle activity, and suppressed the changes in IL-6 protein in plasma. In addition to its inhibition of calcineurin activity, FK506 administration totally suppressed the exercise-induced IL-6 gene transcription, likely by an inhibition of p38 activation. Taken together, these results demonstrate that in addition to p38 MAPK, increased calcineurin activity is one of the signalling events involved in IL-6 gene transcription. PMID- 17133351 TI - Oncogenic KRAS provides a uniquely powerful and variable oncogenic contribution among RAS family members in the colonic epithelium. AB - Activating mutations of the RAS family of small GTPases are among the most common genetic events in human tumorigenesis. Constitutive activation of the three canonical family members, KRAS, NRAS, and HRAS segregate strongly by tissue type. Of these, KRAS mutations predominate in human tumors, including those arising from the colon and lung. We sought to compare the oncogenic contributions of different RAS isoforms in a comparable genetic setting and to explore downstream molecular changes that may explain the apparent differential oncogenic effects of the various RAS family members. We utilized colorectal cancer cell lines characterized by oncogenic KRAS in parallel with isogenically derived lines in which the mutant allele has been disrupted. We additionally attempted to reconstitute the isogenic derivatives with oncogenic forms of other RAS family members and analyze them in parallel. Pairwise analysis of HCT 116 and DLD-1 cell lines as well as their isogenic derivatives reveals distinct K-RAS(G13D) signatures despite the genetic similarities of these cell lines. In DLD-1, for example, oncogenic K-RAS enhances the motility of these cells by downregulation of Rap1 activity, yet is not associated with increased ERK1/2 phosphorylation. In HCT 116, however, ERK1/2 phosphorylation is elevated relative to the isogenic derivative, but Rap1 activity is unchanged. K-RAS is uniquely oncogenic in the colonic epithelium, though the molecular aspects of its oncogenic contribution are not necessarily conserved across cell lines. We therefore conclude that the oncogenic contribution of K-RAS is a function of its multifaceted functionality and is highly context-dependent. PMID- 17133352 TI - Connective tissue growth factor (CTGF/CCN2) is a downstream mediator for TGF beta1-induced extracellular matrix production in osteoblasts. AB - Connective tissue growth factor (CTGF/CCN2) is a cysteine-rich, extracellular matrix (ECM) protein that acts as an anabolic growth factor to regulate osteoblast differentiation and function. Recent studies have identified CTGF as a downstream effector of transforming growth factor-beta1 (TGF-beta1) for certain functions in specific cell types. In this study, we examined the role of CTGF as a downstream mediator of TGF-beta1-induced ECM production and cell growth in osteoblasts. Using primary cultures, we demonstrated that TGF-beta1 is a potent inducer of CTGF expression in osteoblasts, and that this induction occurred at all stages of osteoblast differentiation from the proliferative through mineralization stages. TGF-beta1 treatment of osteoblasts increased the expression and synthesis of the ECM components, collagen and fibronectin. When CTGF-specific siRNA was used to prevent TGF-beta1 induction of CTGF expression, it also inhibited collagen and fibronectin production, thereby demonstrating the requirement of CTGF for their up-regulation. To examine the effects of TGF-beta1 on osteoblast cell growth, cultures were treated with TGF-beta1 during the proliferative stage. Cell number was significantly reduced and the cells exhibited a decrease in G1 cyclin expression, consistent with TGF-beta1-induced cell-cycle arrest. Cultures transfected with CTGF siRNA prior to TGF-beta1 treatment showed an even greater reduction in cell number, suggesting that TGF beta1-induced growth arrest is independent of CTGF in osteoblasts. Collectively, these data demonstrate for the first time that CTGF is an essential downstream mediator for TGF-beta1-induced ECM production in osteoblasts, but these two growth factors function independently regarding their opposing effects on osteoblast proliferation. PMID- 17133353 TI - The role of noggin in human mesenchymal stem cell differentiation. AB - Noggin is a secreted protein that inhibits the binding of bone morphogenetic proteins (BMPs) to their cognate receptor. Its role in human mesenchymal stem cell differentiation has not been well studied. Here, we studied the effect of noggin on human mesenchymal stem cell differentiation induced by inflammatory cytokines (activated T-cell conditioned medium (ACTTCM) or the combination of four T-cell cytokines, TNF-alpha, TGF-beta, IFN-gamma, and IL-17 (TTII)), BMPs, or dexamthasone (DEX). HMSC treated with TTII alone rapidly induced alkaline phosphatase (AlkP) activity. Inclusion of noggin resulted in an additive effect. Noggin acted additively with DEX to induce a significantly higher level of AlkP induction than either noggin or DEX alone. Noggin was examined for its ability to inhibit mineralization in long-term cultures of HMSC stimulated with BMP-2, BMP 6, BMP-7, DEX, or TTII. Surprisingly, noggin alone induced mineralization while it did not inhibit mineralization induced by TTII or BMP-2, BMP-6, or BMP-7. Interestingly, when HMSC were treated with both noggin and DEX they acted synergistically to induce mineralization nearly 3-fold over DEX alone and 30-fold over noggin alone. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed that T-cell cytokines induced noggin, Runx2, BMP-2, and osteocalcin gene expression, while noggin alone induced BMP-2 and osteocalcin gene expression, but not Runx2, although it increased the expression of ActRII, a receptor for BMP-2. These results suggest that in HMSC, the anabolic action of inflammation on bone formation occurs through the induction of noggin, which then induces BMP-2 receptor and BMP-2 leading to the activation of the differentiation process. PMID- 17133354 TI - IGF-1 upregulates electroneutral K-Cl cotransporter KCC3 and KCC4 which are differentially required for breast cancer cell proliferation and invasiveness. AB - The cellular function of electroneutral K-Cl cotransport (KCC) is to regulate epithelial ion transport and osmotic homeostasis. Here we investigate the mechanisms by which insulin-like growth factor 1 (IGF-1) cooperates with KCC to modulate breast cancer biology. IGF-1 stimulates KCC activity of MCF-7 breast cancer cells in a dose- and time-dependent manner. Increased KCC3 and KCC4 abundances contribute to IGF-1-enhanced KCC activity. Endogenous cellular invasiveness was modestly attenuated by KCC4-specific siRNA and the residual invasiveness was much less sensitive to IGF-1 stimulation. KCC3 knockdown significantly reduced basal growth rate and almost abolished IGF-1-stimulated cell proliferation. Consistently, MCF-7 cells obtained advantage in cell proliferation and invasiveness by overexpression of KCC3 and KCC4, respectively. Blockade of gene transcription by actinomycin D abolished IGF-1-mediated increase in KCC3 and KCC4 mRNA, indicating that IGF-1 increases KCC abundance through the regulation of KCC genes. IGF-1 treatment triggered phosphatidylinositol 3-kinase and mitogen-activated protein kinase (MAPK) cascades which were differentially required for IGF-1-stimulated biosynthesis of KCC3 and KCC4. Loss-of-function mutations in KCC significantly inhibited the development and progression of xenograft tumor in SCID mice. The expression level of IGF-1 and KCC polypeptides in the surgical specimens showed a good linear correlation, suggesting autocrine or paracrine IGF-1 stimulation of KCC production in vivo. Among patients with early-stage node-negative breast cancer, disease-free survival (DFS) and overall survival (OS) curves were significantly different based on IGF-1 and KCC expression. Thus, we conclude that KCC activation by IGF-1 plays an important role in IGF-1 receptor signaling to promote growth and spread of breast cancer cells. PMID- 17133355 TI - MAP kinase-dependent, NF-kappaB-independent regulation of inhibitor of apoptosis protein genes by TNF-alpha. AB - The inhibitor of apoptosis protein (IAP) family of molecules regulates apoptotic processes triggered by various stimuli. However, the mechanisms involved in the regulation of the IAP genes are not fully understood. In this report, we examined roles of nuclear factor-kappaB (NF-kappaB) and mitogen-activated protein (MAP) kinases in tumor necrosis factor-alpha (TNF-alpha)-induced expression of IAP genes. In human endothelial cells, TNF-alpha induced c-IAP1 and c-IAP2, but not XIAP and TIAP/Survivin, at the transcriptional level. Inactivation of NF-kappaB by overexpression of a super-repressor mutant of IkappaBalpha did not affect the induction of IAPs by TNF-alpha. In contrast, extracellular signal-regulated kinase, p38 MAP kinase, and c-Jun N-terminal kinase were activated after stimulation with TNF-alpha, and inhibition of each kinase by PD098059, SB203580, curcumin, or SP600125 substantially attenuated the TNF-alpha-induced c-IAP1 and c IAP2 expression. To examine whether the MAP kinases-mediated induction of IAPs contributes to survival of TNF-alpha-exposed cells, cells were pretreated with MAP kinase inhibitors and stimulated with TNF-alpha. Treatment with kinase inhibitors alone did not induce apoptosis but enhanced markedly TNF-alpha triggered apoptosis. Furthermore, overexpression of either c-IAP1 or c-IAP2 diminished the apoptosis-promoting effects of MAP kinase inhibitors. These data indicated that TNF-alpha induced expression of c-IAP1 and c-IAP2 via MAP kinases, but not via NF-kappaB, and that MAP kinases participated in the inhibition of apoptosis by induction of c-IAPs in TNF-alpha-stimulated endothelial cells. PMID- 17133356 TI - Role of atypical protein kinase C isozymes and NF-kappaB in IL-1beta-induced expression of cyclooxygenase-2 in human myometrial smooth muscle cells. AB - Increased myometrial expression of cyclooxygenase-2 (Cox-2) at term results from elevated local levels of inflammatory cytokines, and its inhibition provides a potential route for intervention in human pre-term labor. We have identified a role for atypical protein kinase C (PKC) isozymes in IL-1beta-induced Cox-2 expression in human myometrial smooth muscle cells (HMSMC). The PKC inhibitor GF109203X (10 microM) inhibited IL-1beta-induced Cox-2 protein and RNA expression, which were also reduced by MAPK and nuclear factor kappaB (NF-kappaB) inhibitors. GF109203X did not affect MAPK activities, and neither did it replicate the effect of p38 MAPK inhibition on Cox-2 mRNA stability, suggesting that PKC operates through an independent mechanism. The effect of GF109203X remained intact after depletion of conventional and novel PKC isozymes by phorbol ester pre-treatment. In contrast LY379196 (10 microM), which at micromolar concentrations inhibits all but atypical PKCs, did not affect Cox-2 expression. A peptide corresponding to the pseudosubstrate sequence of atypical PKCs blocked Cox-2 protein expression, whereas the sequence from conventional PKCs was ineffective. GF109203X did not affect NF-kappaB binding to nuclear proteins, but strongly reduced NF-kappaB-dependent transcription in luciferase reporter assays. Our findings indicate that IL-1beta-induced Cox-2 expression in HMSMC in culture requires p38-MAPK-mediated mRNA stabilization and an independent activation of Cox-2 transcription which is dependent on the action of atypical PKCs, probably through direct stimulation of the transactivating activity of NF-kappaB. PMID- 17133357 TI - Development of the terminally differentiated state sensitizes epiphyseal chondrocytes to apoptosis through caspase-3 activation. AB - The maturation of epiphyseal chondrocytes is accompanied by dramatic changes in energy metabolism and shifts in proteins concerned with the induction of apoptosis. We evaluated the role of mitochondria in this process by evaluating the membrane potential (Delta psi m) of chondrocytes of embryonic tibia and the epiphyseal growth plate. We observed that there was a maturation-dependent change in fluorescence, indicating a fall in the Delta psi m. The level of mitochondrial Bcl-2 was decreased during maturation, while in the same time period there was an obvious increase in Bax levels in the mitochondrial fraction of the terminally differentiated chondrocytes. Bcl(xL), another anti-apoptotic protein, was also robustly expressed in the mitochondrial fraction, but its expression was not dependent on the maturation status of the chondrocytes. We found that caspase-3 was present throughout the growth plate and in hypertrophic cells in culture. We blocked caspase-3 activity and found that alkaline phosphatase staining and mineral formation was decreased, and the cells had lost their characteristic shape. Moreover, we noted that the undifferentiated cells were insensitive to elevated concentrations of inorganic phosphate (Pi). It is concluded that during hypertrophy, the change in membrane potential, the increased binding of a pro apoptotic protein to mitochondria, and the activation of caspase-3 serve to prime cells for apoptosis. Only when the terminally differentiated chondrocytes are challenged with low levels of apoptogens there is activation of apoptosis. PMID- 17133358 TI - The role of dynamin 3 in the testis. AB - We report here that dynamin 3 in the testis is associated with structures termed tubulobulbar complexes that internalize intact intercellular junctions during sperm release and turnover of the blood-testis barrier. The protein lies adjacent to an actin-Arp2/3 network that cuffs the double plasma membrane tubular invagination at the core of each complex. To explore the possible relationship between dynamin 3 and nectin-based adhesion junctions, we transiently transfected DsRed-tagged dynamin 3 into MDCK cells stably transfected with eGFP-tagged nectin 2, one of the adhesion molecules known to be expressed in Sertoli cells at adhesion junctions. Cells transfected with the dynamin 3 construct had less uniformly distributed nectin 2 at intercellular contacts when compared to control cells expressing only nectin 2 or transfected with the DsRed plasmid alone. Significantly, tubular extensions positive for nectin 2 were visible projecting into the cells from regions of intercellular contact. Our findings are consistent with the conclusion that dynamin 3 is involved with tubulobulbar morphogenesis. Dynamin 3 also occurs in concentrated deposits around the capitulum and striated columns in the connecting piece of sperm tails suggesting that the protein in these cells may function to stabilize the base of the tail or serve as a reservoir for use during or after fertilization. PMID- 17133359 TI - A possible role of autogenous IFN-beta for cytokine productions in human fibroblasts. AB - It has been already known that human diploid fibroblasts are able to produce not only high levels of IFN-beta but also various kinds of cytokines by poly rI: poly rC, and some inflammatory cytokines are induced by IFN-beta gene activation. We also obtained similar results. However, in our system, cytokine productions were extremely enhanced by treating the cells with a low dose of type 1 IFN and the priming effects on cytokine productions were blocked by cycloheximide similar to those on IFN-beta productions. Most of cytokines were produced later than IFN beta and synthesis patterns of their mRNA showed the same phenomena. We made clear that cytokine productions by poly rI: poly rC are mediated by secreted IFN beta at a protein level using a monoclonal antibody against human IFN-beta. Further, it was shown that intra-cellular IFN-beta which is not secreted might also participate in cytokine productions. Meanwhile, IL-1beta induced various kinds of cytokines in human fibroblasts and production time courses of these cytokines were similar to those of poly rI: poly rC induced cytokines. Although secreted IFN-beta was not detected in IL-1beta stimulated culture, expression of IFN-beta mRNA was augmented. These results showed that priming effects of type 1 IFN on cytokine productions by poly rI: poly rC might not be the direct action, but successive IFN-beta production might be essential in the production processes of other cytokines. Further, it was suggested that inducible IFN-beta might also take part in IL-1beta-induced cytokine productions. PMID- 17133360 TI - CCL20 chemokine induces both osteoblast proliferation and osteoclast differentiation: Increased levels of CCL20 are expressed in subchondral bone tissue of rheumatoid arthritis patients. AB - We evaluated the role of CCL20 (MIP-3alpha) chemokine in cells directly involved in the remodeling of bone tissue (osteoblasts and osteoclasts) and we confirmed its expression in the subchondral bone tissue of rheumatoid arthritis (RA) patients. The expression of CCL20 and of its receptor CCR6 was evaluated in osteoblasts isolated from bone tissue of post-traumatic (PT) patients. Functional tests were performed to evaluate osteoblast proliferation and matrix protein modulation. Immunohistochemical analysis for CCR6, CCL20, and RANKL was performed on bone samples from RA patients. The role of CCL20 was then analyzed in osteoclast differentiation. We found that in basal conditions CCR6, but not its ligand CCL20, was highly expressed by osteoblasts. Functional analysis on osteoblasts showed that CCL20 significantly increased cellular proliferation but did not affect matrix protein expression. Pro-inflammatory cytokines significantly induced the release of CCL20 and RANKL by human osteoblasts but did not modulate CCR6 expression. Increased expression of CCR6, CCL20, and RANKL was confirmed in RA subchondral bone tissue biopsies. We demonstrated that CCL20 was also an earlier inducer of osteoclast differentiation by increasing the number of pre-osteoclasts, thus favoring cell fusion and MMP-9 release. Our results add new insight to the important role of the CCL20/CCR6, RANKL system in the bone tissue of RA. The contemporary action of CCL20 on osteoblasts and osteoclasts involved in the maintenance of bone tissue homeostasis demonstrates the important role of this compartment in the evolution of RA, by showing a clear uncoupling between new bone formation and bone resorption. PMID- 17133361 TI - Bcl-2 expression modulates cell adhesion and migration promoting branching of ureteric bud cells. AB - Bcl-2 is the founding member of a family of proteins that influence apoptosis. During kidney development bcl-2 not only acts as a survival factor, but may also impact cell adhesive mechanisms and by extension branching morphogenesis. The interrelationship between cell adhesion, migration and apoptosis, important during development, is poorly understood. Here we examined the impact lack of bcl 2, an inhibitor of apoptosis, has on ureteric bud (UB) cell adhesion, migration, and branching morphogenesis. Bcl-2 -/- UB cells demonstrated increased cell migration, increased cell invasion and decreased adhesion to vitronectin and fibronectin compared with wild-type cells. Bcl-2 +/+ UB cells readily branched in collagen gel and Matrigel while bcl-2 -/- UB cells did not undergo significant branching in either matrix. Re-expression of bcl-2 in bcl-2 -/- UB cells restored their ability to undergo branching morphogenesis in Matrigel. Consistent with our in vitro data, we show that in the absence of bcl-2, embryonic kidneys undergo decreased UB branching. We observed decreased numbers of UB branch points, UB branch tips and a decreased distance to the first UB branch point in the absence of bcl-2. The alterations in bcl-2 -/- UB cell adhesion and migration was also associated with a significant alteration in expression of a number of extracellular matrix proteins. Bcl-2 -/- UB cells exhibited increased fibronectin expression and decreased thrombospondin-1 and osteopontin expression. Taken together, these data suggest that bcl-2 is required for the proper regulation of cell adhesive and migratory mechanisms, perhaps through modulation of the cellular microenvironment. PMID- 17133362 TI - Potent stimulation of large-conductance Ca2+-activated K+ channels by rottlerin, an inhibitor of protein kinase C-delta, in pituitary tumor (GH3) cells and in cortical neuronal (HCN-1A) cells. AB - The effects of rottlerin, a known inhibitor of protein kinase C-delta activation, on ion currents were investigated in pituitary tumor (GH3) cells. Rottlerin (0.3 100 microM) increased the amplitude of Ca2+-activated K+ current (I K(Ca)) in a concentration-dependent manner with an EC50 value of 1.7 microM. In intracellular perfusion with rottlerin (1 microM) or staurosporine (10 microM), phorbol 12 myristate 13-acetate-induced inhibition of I K(Ca) in these cells was abolished. In cell-attached mode, rottlerin applied on the extracellular side of the membrane caused activation of large-conductance Ca2+-activated K+ (BK(Ca)) channels, and a further application of BAPTA-AM (10 microM) to the bath had no effect on rottlerin-stimulated channel activity. When cells were exposed to rottlerin, the activation curve of these channels was shifted to less positive potential with no change in the slope factor. Rottlerin increased BK(Ca)-channel activity in outside-out patches. Its change in kinetic behavior of BK(Ca) channels is primarily due to an increase in mean open time. With the aid of minimal kinetic scheme, a quantitative description of rottlerin stimulation on BK(Ca) channels in GH3 cells was also provided. Under current-clamp configuration, rottlerin (1 microM) decreased the firing of action potentials. I K(Ca) elicited by simulated action potential waveforms was enhanced by this compound. In human cortical HCN-1A cells, rottlerin (1 microM) could also interact with the BK(Ca) channel to stimulate I K(Ca). Therefore, rottlerin may directly activate BK(Ca) channels in neurons or endocrine cells. PMID- 17133363 TI - Molecular signaling and genetic pathways of senescence: Its role in tumorigenesis and aging. AB - In response to progressive telomere shortening in successive cell divisions, normal somatic cells enter senescence, during which they cease to proliferate irreversibly and undergo dramatic changes in gene expression. Senescence can also be activated by various types of stressful stimuli, including aberrant oncogenic signaling, oxidative stress, and DNA damage. Because of the limited proliferative capacity imposed by senescence, as well as the ability of senescent cells to influence neighboring non-senescent cells, senescence has been proposed to play an important role in tumorigenesis and to contribute to aging. Considerable effort has been put into elucidating the molecular mechanisms of senescence, including the signals that trigger senescence, the molecular pathways by which cells enter senescence, and evidence that supports its role in tumorigenesis and aging. PMID- 17133364 TI - Features that determine telomere homolog oligonucleotide-induced therapeutic DNA damage-like responses in cancer cells. AB - Cancer is the second leading cause of death in the USA, with metastatic disease proving a particular management challenge. Treatment modalities for patients with metastatic disease are limited, and survival beyond 5 years is uncommon. We have reported that an 11-base DNA oligonucleotide 100% homologous to the telomere 3' overhang can induce apoptosis, senescence and/or differentiation of several types of malignant cells in vitro and in vivo, while having minimal effect on normal cells. We now report that 22 oligonucleotides, 9-20 bases in length, with or without a 5' phosphate group and with varying homology (40-100%) to the 3' overhang, inhibit growth and induce apoptosis of human cell lines derived from breast cancers, pancreatic and ovarian carcinomas, and malignant melanoma, lines that lack p53 and/or p16 and harbor a variety of other abnormalities in key regulatory signaling pathways. Cytosine (C) content adversely affected oligonucleotide efficacy, decreasing their effect on cellular apoptosis by > or =80%. These data confirm and expand our earlier work suggesting that such telomere homolog oligonucleotides (T-oligos) target an innate anti-cancer defense system in human cells and may provide an effective treatment for cancers of multiple different cellular origins and genetic profile. PMID- 17133366 TI - Protein probabilities in shotgun proteomics: evaluating different estimation methods using a semi-random sampling model. AB - The calculation of protein probabilities is one of the most intractable problems in large-scale proteomic research. Current available estimating methods, for example, ProteinProphet, PROT_PROBE, Poisson model and two-peptide hits, employ different models trying to resolve this problem. Until now, no efficient method is used for comparative evaluation of the above methods in large-scale datasets. In order to evaluate these various methods, we developed a semi-random sampling model to simulate large-scale proteomic data. In this model, the identified peptides were sampled from the designed proteins and their cross-correlation scores were simulated according to the results from reverse database searching. The simulated result of 18 control proteins was consistent with the experimental one, demonstrating the efficiency of our model. According to the simulated results of human liver sample, ProteinProphet returned slightly higher probabilities and lower specificity than real cases. PROT_PROBE was a more efficient method with higher specificity. Predicted results from a Poisson model roughly coincide with real datasets, and the method of two-peptide hits seems solid but imprecise. However, the probabilities of identified proteins are strongly correlated with several experimental factors including spectra number, database size and protein abundance distribution. PMID- 17133367 TI - Quantitative proteomic analysis of the budding yeast cell cycle using acid cleavable isotope-coded affinity tag reagents. AB - Quantitative profiling of proteins, the direct effectors of nearly all biological functions, will undoubtedly complement technologies for the measurement of mRNA. Systematic proteomic measurement of the cell cycle is now possible by using stable isotopic labeling with isotope-coded affinity tag reagents and software tools for high-throughput analysis of LC-MS/MS data. We provide here the first such study achieving quantitative, global proteomic measurement of a time-course gene expression experiment in a model eukaryote, the budding yeast Saccharomyces cerevisiae, during the cell cycle. We sampled 48% of all predicted ORFs, and provide the data, including identifications, quantitations, and statistical measures of certainty, to the community in a sortable matrix. We do not detect significant concordance in the dynamics of the system over the time-course tested between our proteomic measurements and microarray measures collected from similarly treated yeast cultures. Our proteomic dataset therefore provides a necessary and complementary measure of eukaryotic gene expression, establishes a rich database for the functional analysis of S. cerevisiae proteins, and will enable further development of technologies for global proteomic analysis of higher eukaryotes. PMID- 17133368 TI - The role of periplasmic antioxidant enzymes (superoxide dismutase and thiol peroxidase) of the Shiga toxin-producing Escherichia coli O157:H7 in the formation of biofilms. AB - This study examined the role of the periplasmic oxidative defense proteins, copper, zinc superoxide dismutase (SodC), and thiol peroxidase (Tpx), from the Shiga toxin-producing Escherichia coli O157:H7 (STEC) in the formation of biofilms. Proteomic analyses have shown significantly higher expression levels of both periplasmic antioxidant systems (SodC and Tpx) in STEC cells grown under biofilm conditions than under planktonic conditions. An analysis of their growth phase-dependent gene expression indicated that a high level of the sodC expression occurred during the stationary phase and that the expression of the tpx gene was strongly induced only during the exponential growth phase. Exogenous hydrogen peroxide reduced the aerobic growth of the STEC sodC and tpx mutants by more than that of their parental strain. The two mutants also displayed significant reductions in their attachment to both biotic (HT-29 epithelial cell) and abiotic surfaces (polystyrene and polyvinyl chloride microplates) during static aerobic growth. However, the growth rates of both wild-type and mutants were similar under aerobic growth conditions. The formation of an STEC biofilm was only observed with the wild-type STEC cells in glass capillary tubes under continuous flow-culture conditions compared with the STEC sodC and tpx mutants. To the best of our knowledge, this is the first mutational study to show the contribution of sodC and tpx gene products to the formation of an E. coli O157:H7 biofilm. These results also suggest that these biofilms are physiologically heterogeneous and that oxidative stress defenses in both the exponential and stationary growth stages play important roles in the formation of STEC biofilms. PMID- 17133369 TI - The iron-regulated transcriptome and proteome of Neisseria meningitidis serogroup C. AB - Restricting bacterial growth by iron-chelating proteins that reduce iron availability in mucosal secretions and body fluids belongs to basic mechanisms of innate immunity. Most pathogens and commensals thus developed gene regulons responding to iron concentration and encoding iron acquisition systems and genes involved in host colonization and virulence. Here, we analyzed the steady-state composition of the iron-regulated proteome and transcriptome of an invasive serogroup C clinical isolate of Neisseria meningitidis. The proteome of meningococci grown under iron-depleted and iron-replete conditions was analyzed by 2-DE and proteins exhibiting significantly altered expression were identified by MALDI-TOF MS analysis. In parallel, total RNA was isolated from the same cultures and iron-regulated genes were identified using whole-genome DNA microarrays. The proteome and the transcriptome were found to overlap by only 19 iron-regulated genes/proteins, with 111 genes/proteins being significantly up regulated in iron-replete cultures and 130 genes/proteins being up-regulated during iron starvation, respectively. Comparisons with published transcriptomic data for N. meningitidis serogroup B, moreover, indicate that expression of up to 20% of all meningococcal genes can be subject to regulation in function of iron availability. PMID- 17133370 TI - Proteomics of ischemia and reperfusion injuries in rabbit myocardium with and without intervention by an oxygen-free radical scavenger. AB - A brief period of ischemia followed by timely reperfusion may lead to prolonged, yet reversible, contractile dysfunction (myocardial stunning). Damage to the myocardium occurs not only during ischemia, but also during reperfusion, where a massive release of oxygen-free radicals (OFR) occurs. We have previously utilized 2-DE and MS to define 57 protein spot changes during brief ischemia/reperfusion (15 min ischemia, 60 min reperfusion; 15I/60R) injury in a rabbit model (White, M. Y., Cordwell, S. J., McCarron, H. C. K., Prasan, A. M. et al., Proteomics 2005, 5, 1395-1410) and shown that the majority of these occur because of physical and/or chemical PTMs. In this study, we subjected rabbit myocardium to 15I/60R in the presence of the OFR scavenger N-(2-mercaptopropionyl) glycine (MPG). Thirty-seven of 57 protein spots altered during 15I/60R remained at control levels in the presence of MPG (15I/60R + MPG). Changes to contractile proteins, including myosin light chain 2 (MLC-2) and troponin C (TnC), were prevented by the addition of MPG. To further investigate the individual effects of ischemia and reperfusion, we generated 2-DE gels from rabbit myocardium subjected to brief ischemia alone (15I/0R), and observed alterations of 33 protein spots, including 18/20 seen in both 15I/60R-treated and 15I/60R + MPG treated tissue. The tissue was also subjected to ischemia in the presence of MPG (15I/0R + MPG), and 21 spot changes, representing 14 protein variants, remained altered despite the presence of the OFR scavenger. These ischemia-specific proteins comprised those involved in energy metabolism (lactate dehydrogenase and ATP synthase alpha), redox regulation (NADH ubiquinone oxidoreductase 51 kDa and GST Mu), and stress response (Hsp27 and 70, and deamidated alpha B-crystallin). We conclude that contractile dysfunction associated with myocardial stunning is predominantly caused by OFR damage at the onset of reperfusion, but that OFR independent damage also occurs during ischemia. These ischemia-specific protein modifications may be indicative of early myocardial injury. PMID- 17133371 TI - Protein clusters associated with carcinogenesis, histological differentiation and nodal metastasis in esophageal cancer. AB - We examined the proteomic background of esophageal cancer. We used laser microdissection to obtain tumor tissues from 72 esophageal squamous cell carcinoma cases and adjacent normal tissues in 57 of these cases. The 2D-DIGE generated quantitative expression profiles with 1730 protein spots. Based on the intensity of the protein spots, unsupervised classification distinguished the tumor tissues from their normal counterparts, and subdivided the tumor tissues according to their histological differentiation. We identified 498 protein spots with altered intensity in the tumor tissues, which protein identification by LC MS/MS showed to correspond to 217 gene products. We also found 41 protein spots that were associated with nodal metastasis, and identified 33 proteins corresponding to the spots, including cancer-associated proteins such as alpha actinin 4, hnRNP K, periplakin, squamous cell carcinoma antigen 1 and NudC. The identified cancer-associated proteins have been previously reported to be individually involved in a range of cancer types, and our study observed them collectively in a single type of malignancy, esophageal cancer. As the identified proteins are involved in important biological processes such as cytoskeletal/structural organization, transportation, chaperon, oxidoreduction, transcription and signal transduction, they may function in a coordinate manner in carcinogenesis and tumor progression of esophageal cancer. PMID- 17133372 TI - Proteome analysis of bronchoalveolar lavage in lung diseases. AB - The proteomic approach is complementary to genomics and enables protein composition to be investigated under various clinical conditions. Its application to the study of bronchoalveolar lavage (BAL) is extremely promising. BAL proteomic studies were initially based on two-dimensional electrophoretic separation of complex protein samples and subsequent identification of proteins by different methods. With the techniques available today it is possible to attain many different research objectives. BAL proteomics can contribute to the identification of proteins in alveolar spaces with possible insights into pathogenesis and clinical application for diagnosis, prognosis and therapy. Many proteins with different functions have already been identified in BAL. Some could be biomarkers that need to be individually confirmed by correlation with clinical parameters and validation by other methods on larger cohorts of patients. The standardization of BAL sample preparation and processing for proteomic studies is an important goal that would promote and facilitate clinical applications. Here, we review the principal literature on BAL proteomic analysis applied to the study of lung diseases. PMID- 17133373 TI - Quantitative protein analysis from formalin-fixed tissues: implications for translational clinical research and nanoscale molecular diagnosis. AB - Owing to its cross-linking effects, it is currently believed that formalin fixation of routinely processed tissues in the clinic prevents protein extraction and profiling. The aim of our study was to develop a robust, fast, standardized, and easy to use technique for the solubilization of non-degraded, full length, and immunoreactive proteins from formalin-fixed tissues for western blot and protein microarray analysis. Sections of routinely processed formalin-fixed and paraffin-embedded tissues of various origin were analysed. After deparaffination, tissues were manually dissected from the slides and transferred into an optimized protein extraction buffer system. Proteins were solubilized and subsequently analysed by western blot and reverse phase protein microarrays. We succeeded in isolating non-degraded, soluble, and immunoreactive proteins from routinely processed formalin-fixed tissues. We were able to detect membrane, cytoplasmic and nuclear proteins at the expected molecular weight. No differences were found in the protein yield and protein abundances between fresh frozen and formalin fixed tissues. Using western blots and reverse phase protein microarrays, the receptor tyrosine kinase HER2, an important protein target for antibody based cancer treatment, was reliably measured in formalin-fixed breast cancer biopsy samples when compared with measurement by immunohistochemistry and fluorescence in situ hybridization; remarkably, immunohistochemically equivocal cases (score 2+) can be categorized according to HER2 protein abundance. Our new clinically orientated multiplexed protein measurement system may be generally applicable to determine the relative abundances of known disease-related proteins in small amounts of routinely processed formalin-fixed tissue samples for research and diagnosis. This technique may also be used to identify, characterize, and validate known and new protein markers in a variety of human diseases. PMID- 17133378 TI - Granulocyte- and monocyte-platelet adhesion index in coronary and peripheral blood after extracorporeal circulation and reperfusion. AB - BACKGROUND: Neutrophil-granulocyte and mononuclear-cell functional changes occur during cardiopulmonary bypass and cardiovascular surgery. In particular, leukocyte-platelet interaction, leading to generation of heterotypic coaggregates, represents an amplification mechanism of the local inflammatory response and tissue damage. METHODS: Samples of 20 patients were drawn from venous coronary sinus before cardioplegic arrest and immediately after reperfusion, as well as from peripheral blood at 5 and 24 h postoperatively. The granulocyte and monocyte surface expression of CD162, CD15s, CD18, and CD11b were quantified by flow cytometry at the different times. Parallel variations of circulating leukocyte-platelet conjugates (percentages) and a derived (cell number-normalized) leukocyte-platelet adhesion index were measured using a combination of antibodies against CD45, CD14, and CD41a. The evaluation of platelet functional state was carried out using antibodies against CD62P (P selectin) and PAC-1. RESULTS: Monocyte and granulocyte cell number increased markedly in coronary blood at reperfusion and in peripheral blood postoperatively when compared with measurements done before cardioplegia. A very different course characterized the changes of the leukocyte-platelet adhesion index with respect to the variations of circulating leukocyte-platelet coaggregates (percentages). Leukocyte molecules expression showed no significant variations for CD15s on both the leukocyte subsets, while a significant up-modulation for CD162 was observed on monocytes at 24 h after extracorporeal circulation (P = 0.0002), and for CD11b on granulocytes at 5 h postoperatively (P = 0.033). A loss of CD162 expression was observed in coronary blood at reperfusion (P = 0.0038) on granulocytes, associated to a down-modulation of CD18 (P = 0.0033) and CD11b (P = 0.0184) in peripheral blood at 24 h postoperatively. No significant up-regulation of platelet activatory molecules expression was found at coronary reperfusion, as well as postoperatively in the peripheral blood, when compared with the before cardioplegia derived data. CONCLUSIONS: The over time variations of a normalized leukocyte-platelet adhesion index seem to reflect the cumulative leukocyte platelet functional interaction more accurately than the parallel measurements of cellular conjugates. The absence of platelet activation suggests that the leukocyte membrane modifications play a main role in controlling the formation and stability of heterotypic leukocyte-platelet coaggregates after cardiac surgery with extracorporeal circulation. PMID- 17133379 TI - "Virtual flow cytometry" of immunostained lymphocytes on microscopic tissue slides: iHCFlow tissue cytometry. AB - BACKGROUND: A method and approach is developed for fully automated measurements of immunostained lymphocytes in tissue sections by means of digital color microscopy and patent pending advanced cell analysis. The validation data for population statistic measurements of immunostained lymphocytes in tissue sections using tissue cytometry (TC) is presented. The report is the first to describe the conversion of immunohistochemistry (IHC) data to a flow cytometry-like two parameter dot-plot display, hence the technique is also a virtual flow cytometry. We believe this approach is a paradigm shift, as well as novel, and called the system iHCFlow TC. Seven issues related to technical obstacles to virtual flow cytometry (FC) are identified. DESIGN: Segmentation of a 512 x 474 RGB image and tabular display of statistical results table took 12-15 s using proprietary developed algorithms. We used a panel of seven antibodies for validation on 14 cases of mantle cell lymphoma giving percentage positive, total lymphocytes, and staining density. A total of 2,027 image frames with 810,800 cell objects (COBs) were evaluated. Antibodies to CD3, CD4, CD8, Bcl-1, Ki-67, CD20, CD5 were subjected to virtual FC on tissue. The results of TC were compared with manual counts of expert observers and with the results of flow cytometric immunophenotyping of the same specimen. RESULTS: The correlation coefficient and 95% confidence interval by linear regression analysis yielded a high concordance between manual human results (M), FC results, and TC results per antibody, (r = 0.9365 M vs. TC, r= 0.9537 FC vs. TC). The technical issues were resolved and the solutions and results were evaluated and presented. CONCLUSION: These results suggest the new technology of TC by iHCFlow could be a clinically valid surrogate for both M and FC analysis when only tissue IHC is available for diagnosis and prognosis. The application for cancer diagnosis, monitoring, and prognosis is for objective, rapid, automated counting of immunostained cells in tissues with percentage results. We report a new paradigm in TC that converts IHC staining of lymphocytes to automated results and a flow cytometry-like report. The dot plot histogram display is familiar, intuitive, informative, and provides the pathologists with an automated tool to rapidly characterize the staining and size distribution of the immunoreactive as well as the negative cell population in the tissue. This systems tool is a major improvement over existing ones and satisfies fully the criteria to perform Cytomics (Ecker and Tarnok, Cytometry A 2005;65:1; Ecker and Steiner, Cytometry A 2004;59:182-190; Ecker et al., Cytometry A 2004;59:172-181). PMID- 17133380 TI - Abstracts of the XIIth International Symposium on Luminescence Spectrometry Detection Techniques in Biomedical, Environmental and Food Analysis (ISLS 2006). PMID- 17133381 TI - High-resolution fMRI investigation of the medial temporal lobe. AB - The medial temporal lobe (MTL) is critical for declarative memory formation. Several theories of MTL function propose functional distinctions between the different structures of the MTL, namely the hippocampus and the surrounding cortical areas. Furthermore, computational models and electrophysiological studies in animals suggest distinctions between the subregions of the hippocampus itself. Standard fMRI resolution is not sufficiently fine to resolve activity on the scale of hippocampal subregions. Several approaches to scanning the MTL at high resolutions have been made, however there are limitations to these approaches, namely difficulty in conducting group-level analyses. We demonstrate here techniques for scanning the MTL at high resolution and analyzing the high resolution fMRI data at the group level. To address the issue of cross participant alignment, we employ the ROI-LDDMM alignment technique, which is demonstrated to result in smaller alignment errors when compared with several other common normalization techniques. Finally, we demonstrate that the pattern of activation obtained in the high-resolution functional data is similar to that obtained at lower resolution, although the spatial extent is smaller and the percent signal change is greater. This difference in the pattern of activation may be due to less partial volume sampling in the high-resolution data, resulting in more accentuated regions of activation. PMID- 17133382 TI - Improving functional magnetic resonance imaging motor studies through simultaneous electromyography recordings. AB - Specially designed optoelectronic and data postprocessing methods are described that permit electromyography (EMG) of muscle activity simultaneous with functional MRI (fMRI). Hardware characterization and validation included simultaneous EMG and event-related fMRI in 17 healthy participants during either ankle (n = 12), index finger (n = 3), or wrist (n = 2) contractions cued by visual stimuli. Principal component analysis (PCA) and independent component analysis (ICA) were evaluated for their ability to remove residual fMRI gradient induced signal contamination in EMG data. Contractions of ankle tibialis anterior and index finger abductor were clearly distinguishable, although observing contractions from the wrist flexors proved more challenging. To demonstrate the potential utility of simultaneous EMG and fMRI, data from the ankle experiments were analyzed using two approaches: 1) assuming contractions coincided precisely with visual cues, and 2) using EMG to time the onset and offset of muscle contraction precisely for each participant. Both methods produced complementary activation maps, although the EMG-guided approach recovered more active brain voxels and revealed activity better in the basal ganglia and cerebellum. Furthermore, numerical simulations confirmed that precise knowledge of behavioral responses, such as those provided by EMG, are much more important for event related experimental designs compared to block designs. This simultaneous EMG and fMRI methodology has important applications where the amplitude or timing of motor output is impaired, such as after stroke. PMID- 17133383 TI - Real-time fMRI using brain-state classification. AB - We have implemented a real-time functional magnetic resonance imaging system based on multivariate classification. This approach is distinctly different from spatially localized real-time implementations, since it does not require prior assumptions about functional localization and individual performance strategies, and has the ability to provide feedback based on intuitive translations of brain state rather than localized fluctuations. Thus this approach provides the capability for a new class of experimental designs in which real-time feedback control of the stimulus is possible-rather than using a fixed paradigm, experiments can adaptively evolve as subjects receive brain-state feedback. In this report, we describe our implementation and characterize its performance capabilities. We observed approximately 80% classification accuracy using whole brain, block-design, motor data. Within both left and right motor task conditions, important differences exist between the initial transient period produced by task switching (changing between rapid left or right index finger button presses) and the subsequent stable period during sustained activity. Further analysis revealed that very high accuracy is achievable during stable task periods, and that the responsiveness of the classifier to changes in task condition can be much faster than signal time-to-peak rates. Finally, we demonstrate the versatility of this implementation with respect to behavioral task, suggesting that our results are applicable across a spectrum of cognitive domains. Beyond basic research, this technology can complement electroencephalography-based brain computer interface research, and has potential applications in the areas of biofeedback rehabilitation, lie detection, learning studies, virtual reality-based training, and enhanced conscious awareness. PMID- 17133384 TI - The interaction between orthographic and phonological information in children: an fMRI study. AB - We examined the neural representations of orthographic and phonological processing in children, while manipulating the consistency between orthographic and phonological information. Participants, aged 9-15, were scanned while performing rhyming and spelling judgments on pairs of visually presented words. The orthographic and phonological similarity between words in the pair was independently manipulated, resulting in four conditions. In the nonconflicting conditions, both orthography and phonology of the words were either (1) similar (lime-dime) or (2) different (staff-gain); in conflicting conditions, words had (3) similar phonology and different orthography (jazz-has) or (4) different phonology and similar orthography (pint-mint). The comparison between tasks resulted in greater activation for the rhyming task in bilateral inferior frontal gyri (BA 45/47), and greater activation for the spelling task in bilateral inferior/superior parietal lobules (BA 40/7), suggesting greater involvement of phonological and semantic processing in the rhyming task, and nonlinguistic spatial processing in the spelling task. Conflicting conditions were more difficult in both tasks and resulted in greater activation in the above regions. The results suggest that when children encounter inconsistency between orthographic and phonological information they show greater engagement of both orthographic and phonological processing. PMID- 17133385 TI - Temporal lobe interictal epileptic discharges affect cerebral activity in "default mode" brain regions. AB - A cerebral network comprising precuneus, medial frontal, and temporoparietal cortices is less active both during goal-directed behavior and states of reduced consciousness than during conscious rest. We tested the hypothesis that the interictal epileptic discharges affect activity in these brain regions in patients with temporal lobe epilepsy who have complex partial seizures. At the group level, using electroencephalography-correlated functional magnetic resonance imaging in 19 consecutive patients with focal epilepsy, we found common decreases of resting state activity in 9 patients with temporal lobe epilepsy (TLE) but not in 10 patients with extra-TLE. We infer that the functional consequences of TLE interictal epileptic discharges are different from those in extra-TLE and affect ongoing brain function. Activity increases were detected in the ipsilateral hippocampus in patients with TLE, and in subthalamic, bilateral superior temporal and medial frontal brain regions in patients with extra-TLE, possibly indicating effects of different interictal epileptic discharge propagation. PMID- 17133386 TI - Abnormal activation of the social brain during face perception in autism. AB - ASD involves a fundamental impairment in processing social-communicative information from faces. Several recent studies have challenged earlier findings that individuals with autism spectrum disorder (ASD) have no activation of the fusiform gyrus (fusiform face area, FFA) when viewing faces. In this study, we examined activation to faces in the broader network of face-processing modules that comprise what is known as the social brain. Using 3T functional resonance imaging, we measured BOLD signal changes in 10 ASD subjects and 7 healthy controls passively viewing nonemotional faces. We replicated our original findings of significant activation of face identity-processing areas (FFA and inferior occipital gyrus, IOG) in ASD. However, in addition, we identified hypoactivation in a more widely distributed network of brain areas involved in face processing [including the right amygdala, inferior frontal cortex (IFC), superior temporal sulcus (STS), and face-related somatosensory and premotor cortex]. In ASD, we found functional correlations between a subgroup of areas in the social brain that belong to the mirror neuron system (IFC, STS) and other face-processing areas. The severity of the social symptoms measured by the Autism Diagnostic Observation Schedule was correlated with the right IFC cortical thickness and with functional activation in that area. When viewing faces, adults with ASD show atypical patterns of activation in regions forming the broader face processing network and social brain, outside the core FFA and IOG regions. These patterns suggest that areas belonging to the mirror neuron system are involved in the face-processing disturbances in ASD. PMID- 17133387 TI - Hemodynamic responses in neural circuitries for detection of visual target and novelty: An event-related fMRI study. AB - The oddball paradigm examines attentional processes by establishing neural substrates for target detection and novelty. Event-related functional imaging enables characterization of hemodynamic changes associated with these processes. We studied 36 healthy participants (17 men) applying a visual oddball event related design at 4 Tesla, and performed an unbiased determination of the hemodynamic response function (HRF). Targets were associated with bilateral, albeit leftward predominant changes in frontal-parietal temporal and occipital cortices, and limbic and basal ganglia regions. Activation to novelty was more posteriorly distributed, and frontal activation occurred only on the right, while robust activation was seen in occipital regions bilaterally. Overlapping regions were left thalamus, caudate and cuneus and right parietal precuneus. While robust HRFs characterized most regions, target detection was associated with a negative HRF in the right parietal precuneus and a biphasic HRF in thalamus, basal ganglia, and all occipital regions. Both height of the HRF and longer time to peak in the right cingulate were associated with slower response time. Sex differences were observed, with higher HRF peaks for novelty in men in right occipital regions, and longer time to peak in the left hemisphere. Age was associated with reduced peak HRF in left frontal region. Thus, indices of the HRF can be used to better understand the relationship between hemodynamic changes and performance and can be sensitive to individual differences. PMID- 17133388 TI - Neural correlates of automatic beliefs about gender and race. AB - Functional MRI was used to identify the brain areas underlying automatic beliefs about gender and race, and suppression of those attitudes. Participants (n = 20; 7 females) were scanned at 3 tesla while performing the Implicit Association Test (IAT), an indirect measure of race and gender bias. We hypothesized that ventromedial prefrontal cortex areas (PFC) would mediate gender and racial stereotypic attitudes, and suppression of these beliefs would recruit dorsolateral prefrontal cortex (DLPFC) and the anterior cingulate cortex (ACC). Performance data on the IAT revealed gender and racial biases. Racial bias was correlated with an explicit measure of racism. Results showed activation of anteromedial PFC and rostral ACC while participants implicitly made associations consistent with gender and racial biases. In contrast, associations incongruent with stereotypes recruited DLPFC. Implicit gender bias was correlated with amygdala activation during stereotypic conditions. Results suggest there are dissociable roles for anteromedial and dorsolateral PFC circuits in the activation and inhibition of stereotypic attitudes. PMID- 17133389 TI - A glimpse into your vision. AB - Invasive recordings of local field potentials (LFPs) have been used to "read the mind" of monkeys in real time. Here we investigated whether noninvasive field potentials estimated from the scalp-recorded electroencephalogram (EEG) using the ELECTRA source localization algorithm could provide real-time decoding of mental states in healthy humans. By means of pattern recognition techniques on 500-ms EEG epochs, we were able to discriminate accurately from single trials which of four categories of visual stimuli the subjects were viewing. Our results show that it is possible to reproduce the decoding accuracy previously obtained in animals with invasive recordings. A comparison between the decoding results and the subjects' behavioral performance indicates that oscillatory activity (OA), elicited in specific brain regions) codes better for the visual stimulus category presented than the subjects' actual response, i.e., is insensitive to voluntary or involuntary errors. The identification of brain regions participating in the decoding process allowed us to construct 3D-functional images of the task-related OA. These images revealed the activation of brain regions known for their involvement in the processing of this type of visual stimuli. Electrical neuroimaging therefore appears to have the potential to establish what the brain is processing while the stimuli are being seen or categorized, i.e., concurrently with sensory-perceptual processes. PMID- 17133390 TI - Altered functional connectivity in early Alzheimer's disease: a resting-state fMRI study. AB - Previous studies have led to the proposal that patients with Alzheimer's disease (AD) may have disturbed functional connectivity between different brain regions. Furthermore, recent resting-state functional magnetic resonance imaging (fMRI) studies have also shown that low-frequency (<0.08 Hz) fluctuations (LFF) of the blood oxygenation level-dependent signals were abnormal in several brain areas of AD patients. However, few studies have investigated disturbed LFF connectivity in AD patients. By using resting-state fMRI, this study sought to investigate the abnormal functional connectivities throughout the entire brain of early AD patients, and analyze the global distribution of these abnormalities. For this purpose, the authors divided the whole brain into 116 regions and identified abnormal connectivities by comparing the correlation coefficients of each pair. Compared with healthy controls, AD patients had decreased positive correlations between the prefrontal and parietal lobes, but increased positive correlations within the prefrontal lobe, parietal lobe, and occipital lobe. The AD patients also had decreased negative correlations (closer to zero) between two intrinsically anti-correlated networks that had previously been found in the resting brain. By using resting-state fMRI, our results supported previous studies that have reported an anterior-posterior disconnection phenomenon and increased within-lobe functional connectivity in AD patients. In addition, the results also suggest that AD may disturb the correlation/anti-correlation effect in the two intrinsically anti-correlated networks. PMID- 17133391 TI - Regulation of emotional responses elicited by threat-related stimuli. AB - The capacity to voluntarily regulate emotions is critical for mental health, especially when coping with aversive events. Several neuroimaging studies of emotion regulation found the amygdala to be a target for downregulation and prefrontal regions to be associated with downregulation. To characterize the role of prefrontal regions in bidirectional emotion regulation and to investigate regulatory influences on amygdala activity and peripheral physiological measures, a functional magnetic resonance imaging (fMRI) study with simultaneous recording of self-report, startle eyeblink, and skin conductance responses was carried out. Subjects viewed threat-related pictures and were asked to up- and downregulate their emotional responses using reappraisal strategies. While startle eyeblink responses (in successful regulators) and skin conductance responses were amplified during upregulation, but showed no consistent effect during downregulation, amygdala activity was increased and decreased according to the regulation instructions. Trial-by-trial ratings of regulation success correlated positively with activity in amygdala during upregulation and orbitofrontal cortex during downregulation. Downregulation was characterized by left-hemispheric activation peaks in anterior cingulate cortex, dorsolateral prefrontal cortex, and orbitofrontal cortex and upregulation was characterized by a pattern of prefrontal activation not restricted to the left hemisphere. Further analyses showed significant overlap of prefrontal activation across both regulation conditions, possibly reflecting cognitive processes underlying both up- and downregulation, but also showed distinct activations in each condition. The present study demonstrates that amygdala responses to threat-related stimuli can be controlled through the use of cognitive strategies depending on recruitment of prefrontal areas, thereby changing the subject's affective state. PMID- 17133392 TI - Taxing working memory with syntax: bihemispheric modulations. AB - Motivated by claims that relegate the syntactic functions of Broca's region to working memory (WM) and not to language-specific mechanisms, we conducted an fMRI and an aphasia study that featured two varieties of intrasentential dependency relations: One was syntactic movement (e.g., Which boy does the girl think [symbol in text] examined Steven?), the other was antecedent-reflexive binding (e.g., Jill thinks the boy examined himself). In both, WM is required to link two nonadjacent positions. Syntactically, they are governed by distinct rule systems. In health, the two dependencies modulated activity in distinct brain regions within the left inferior frontal gyrus and the left middle temporal gyrus. Binding uniquely modulated activation in the right frontal lobe. Receptive abilities in brain damaged patients likewise distinguished among these syntactic types. The results indicate that sentence comprehension is governed by syntactically carved neural chunks and provide hints regarding a language related region in the right hemisphere. PMID- 17133393 TI - Interparticipant correlations: a model free FMRI analysis technique. AB - FMRI analysis techniques can be broadly divided into model based and data driven techniques. The most widely used approach assumes an explicit temporal hemodynamic model based upon the experimental paradigm. Such an approach has proven very useful and powerful even though it is limited by the accuracy of the prespecified model. An alternative approach is to use data driven techniques like independent component analysis or fuzzy cluster analysis. These approaches have proven useful for exploratory analysis in a multivariate sense; however, they can present additional difficulties in the interpretation of the results. An alternative to these approaches is to take advantage of similarities in the patterns of the hemodynamics between participants [i.e., interparticipant correlation (IPC)]. This FMRI analysis technique enjoys the parsimony of the general linear model (GLM) but does not assume a specific FMRI time course. The technique consists of calculating voxel-wise correlations between participants resulting in IPC maps, which indicate the activated regions the participants have in common. We applied the IPC approach to data collected from healthy controls in an auditory oddball task. As expected, high inter-participant correlations were detected in auditory cortical regions in the temporal lobes where highest correlations were evident. In addition, areas that appear to be involved in the task were detected using IPC's but not the GLM regression. This technique, designed to have increased sensitivity to inter-subject correlations that are not necessarily task-related, may potentially be useful as a compliment to model based approaches. PMID- 17133394 TI - Corpus callosal connection mapping using cortical gray matter parcellation and DT MRI. AB - Population maps of the corpus callosum (CC) and cortical lobe connections were generated by combining cortical gray matter parcellation with the diffusion tensor fiber tractography of individual subjects. This method is based on the fact that the cortical lobes of both hemispheres are interconnected by the corpus callosal fibers. T1-weighted structural MRIs and diffusion tensor MRIs (DT-MRI) of 22 right-handed, healthy subjects were used. Forty-seven cortical parcellations in the dorsal prefrontal cortex, ventral prefrontal cortex, sensory motor cortex, parietal cortex, temporal cortex, and occipital cortex were semi automatically derived from structural MRIs, registered to DT-MRI, and used to identify callosal fibers. The probabilistic connections to each cortex were mapped on entire mid-sagittal CC voxels that had anatomical homology between subjects as determined by spatial registration. According to the population maps of the callosal connections, the ventral prefrontal cortex and parts of the dorsal prefrontal cortex both project fibers through the genu and rostrum. The CC regions through which the superior frontal cortex passes extend into the posterior body. Fibers arising from the parietal lobe and occipital lobe run mainly through the splenium, while fibers arising from the sensory-motor cortex pass through the isthmus. In general, dorsal or medial cortical lobes project fibers through the dorsal region of the CC, while lateral cortical lobes project fibers through the ventral region of the CC. The probabilistic subdivision of the CC by connecting cortical gray matter provides a more precise understanding of the CC. PMID- 17133395 TI - Evaluation of PCA and ICA of simulated ERPs: Promax vs. Infomax rotations. AB - Independent components analysis (ICA) and principal components analysis (PCA) are methods used to analyze event-related potential (ERP) and functional imaging (fMRI) data. In the present study, ICA and PCA were directly compared by applying them to simulated ERP datasets. Specifically, PCA was used to generate a subspace of the dataset followed by the application of PCA Promax or ICA Infomax rotations. The simulated datasets were composed of real background EEG activity plus two ERP simulated components. The results suggest that Promax is most effective for temporal analysis, whereas Infomax is most effective for spatial analysis. Failed analyses were examined and used to devise potential diagnostic strategies for both rotations. Finally, the results also showed that decomposition of subject averages yield better results than of grand averages across subjects. PMID- 17133396 TI - Complex relationship between BOLD signal and synchronization/desynchronization of human brain MEG oscillations. AB - Functional magnetic resonance imaging (fMRI) depends on the coupling of cerebral blood flow, energy demand, and neural activity. The precise nature of this interaction, however, is poorly understood. A positive correlation between BOLD response and cortically generated local field potentials, which reflect the weighted average of synchronized dentrosomatic components of pyramidal synaptic signals, has been demonstrated. Likewise, positive BOLD-responses have been reported in conjunction with scalp-recorded synchronized electromagnetic activity by a number of groups. However, it is not yet clear how the opposite electromagnetic pattern, i.e. cortical desynchronization, is related to the BOLD signal. To address this question, we conducted a combined event-related fMRI and 275 sensor whole-head MEG study during identical visual two-choice reaction time task conditions in 10 human subjects. We found complex sequences of MEG synchronization and desynchronization across a wide frequency range in the visual and motor area in close correspondence with "locales" of positive BOLD-responses. These results indicate that a correspondence of positive BOLD-responses is not exclusively found for cortical synchronization but also for desynchronization, suggesting that the relationship between BOLD signals and electromagnetic activity might be more complex than previously thought. PMID- 17133397 TI - fMRI study of effort and information processing in a working memory task. AB - It is unclear how effort translates into brain function. In this study we endeavored to identify the activity in a working memory task that is related to the allocation of mental resources. Such activity, if present, would be a likely candidate to explain how effort works in terms of brain function. Eleven healthy participants performed a Sternberg task with a memory-set of one, three, or five consonants in an fMRI study. Probe stimuli were either one consonant or one digit. We expected digits to be processed automatically and consonants to require working memory. Because the probe type was unpredictable and subjects had to respond as fast as possible, we expected subjects to allocate mental resources on the basis of the memory-set size, not the probe type. Accordingly, we anticipated that activity in regions involved in effort would be a function of the size of the memory-set, but independent of the type of probe. We found that the reaction time for digits increased in line with our expectation of automatic processing and the reaction time for letters increased in line with our expectation of controlled processing. fMRI revealed that activity in the right ventral prefrontal cortex changed as a function of effort. The ventral anterior cingulate cortex and hypothalamus showed reduced activity as a function of effort. Activity in regions regarded as pivotal for working memory (among others, the left dorsolateral prefrontal cortex, anterior cingulate cortex) appeared to be predominantly related to information processing and not involved in effort. PMID- 17133398 TI - Guided saccades modulate object and face-specific activity in the fusiform gyrus. AB - We investigated the influence of saccadic eye movements on the magnitude of functional MRI (fMRI) activation in brain regions known to participate in object and face perception. In separate runs, subjects viewed a static image of a uniform gray field, a face, or a flower. Every 500 ms a small fixation cross made a discrete jump within the image and subjects were required to make a saccade and fixate the cross at its new location. Each run consisted of alternating blocks in which the subject was guided to make small and large saccades. A comparison of large vs. small saccade blocks revealed robust activity in the oculomotor system, particularly within the frontal eye fields (FEF), intraparietal sulcus (IPS), and superior colliculi regardless of the background image. Activity within portions of the ventral occipitotemporal cortex (VOTC) including the lingual and fusiform gyri was also modulated by saccades, but here saccade-related activity was strongly influenced by the background image. Activity within the VOTC was strongest when large saccadic eye movements were made over an image of a face or a flower compared to a uniform gray image. Of most interest was activity in the functionally predefined face-specific region of the fusiform gyrus, where large saccades made over a face increased activity, but where similar large saccades made over a flower or a uniform gray field did not increase activity. These results demonstrate the potentially confounding influence of uncontrolled eye movements for neuroimaging studies of face and object perception. PMID- 17133399 TI - Neural characteristics of successful and less successful speech and word learning in adults. AB - A remarkable characteristic of the human nervous system is its ability to learn to integrate novel (foreign) complex sounds into words. However, the neural changes involved in how adults learn to integrate novel sounds into words and the associated individual differences are largely unknown. Unlike English, most languages of the world use pitch patterns to mark individual word meaning. We report a study assessing the neural correlates of learning to use these pitch patterns in words by English-speaking adults who had no previous exposure to such usage. Before and after training, subjects discriminated pitch patterns of the words they learned while blood oxygenation levels were measured using fMRI. Subjects who mastered the learning program showed increased activation in the left posterior superior temporal region after training, while subjects who plateaued at lower levels showed increased activation in the right superior temporal region and right inferior frontal gyrus, which are associated with nonlinguistic pitch processing, and prefrontal and medial frontal areas, which are associated with increased working memory and attentional efforts. Furthermore, we found brain activation differences even before training between the two subject groups, including the superior temporal region. These results demonstrate an association between range of neural changes and degrees of language learning, specifically implicating the physiologic contribution of the left dorsal auditory cortex in learning success. PMID- 17133400 TI - Bayesian comparison of spatially regularised general linear models. AB - In previous work (Penny et al., [2005]: Neuroimage 24:350-362) we have developed a spatially regularised General Linear Model for the analysis of functional magnetic resonance imaging data that allows for the characterisation of regionally specific effects using Posterior Probability Maps (PPMs). In this paper we show how it also provides an approximation to the model evidence. This is important as it is the basis of Bayesian model comparison and provides a unified framework for Bayesian Analysis of Variance, Cluster of Interest analyses and the principled selection of signal and noise models. We also provide extensions that implement spatial and anatomical regularisation of noise process parameters. PMID- 17133401 TI - Object identification and lexical/semantic access in children: a functional magnetic resonance imaging study of word-picture matching. AB - Theoretical models for lexical access to visual objects have been based mainly on adult data. To investigate the developmental aspects of object recognition and lexical access in children, a large-scale functional MRI (fMRI) study was performed in 283 normal children ages 5-18 using a word-picture matching paradigm in which children would match an aurally presented noun to one of two pictures (line drawings). Using group Independent Component Analysis (ICA), six task related components were detected, including (a) the posterior superior temporal gyrus bilaterally; (b) the fusiform, inferior temporal, and middle occipital gyri bilaterally; (c) the dorsal aspect of the inferior frontal gyrus bilaterally, the left precuneus, the left superior/middle temporal gyrus, and the anterior cingulate; (d) the right medial fusiform gyrus; (e) a left-lateralized component including the inferior/middle frontal, middle temporal, medial frontal, and angular gyri, as well as the thalamus and the posterior cingulate; and (f) the ventral/anterior aspect of the inferior frontal gyrus bilaterally. Increased activation associated with age was seen in the components (b) and (d) (ventral visual pathway) for object recognition, and (c) and (f) likely associated with semantic maintenance and response selection. Increased activation associated with task performance was seen in components (b) and (d) (ventral visual pathway) while decreased activation associated with task performance was seen in component (f) (ventral/anterior inferior frontal gyrus). The results corroborate the continued development of the ventral visual pathway throughout the developmental period. PMID- 17133402 TI - 'Prefrontal' cognitive performance of healthy subjects positively correlates with cerebral FDOPA influx: an exploratory [18F]-fluoro-L-DOPA-PET investigation. AB - Dopamine neurotransmission influences those cognitive processes, which are generally regarded as prefrontal cortical functions. In previous positron emission-tomography (PET) studies, net blood-brain clearance of [18F]-fluoro-l DOPA (FDOPA) correlated with impaired cognitive performance in patients with Parkinson's disease or schizophrenia. We hypothesized that FDOPA influx also correlates with performance of cognitive tasks associated with prefrontal functioning in healthy volunteers. The net blood-brain clearance of FDOPA (K(in)(app)) was mapped in a group of 11 healthy volunteers and calculated in striatal volumes-of-interest. The Wisconsin-Card-Sorting-Test (WCST), Stroop Test, Trail-Making-Test (TMT-A/B), and Continuous-Performance-Test (CPT-M) had been administered previously to the same subjects. No correlation of K(in) (app) with perseverative errors in WCST or age could be found. However, there were significant positive correlations between the magnitude of K(in)(app) in caudate nucleus, putamen, and midbrain with performance of the TMT-B, CPT-M, and the Stroop test. Highest correlations were found between the time needed to perform the Stroop interference task and the K(in)(app) of striatal areas (Caudate nucleus: -0.780, P = 0.005; putamen: -0.870, P < 0. 001). Thus, the present findings reveal a strong correlation between dopamine synthesis capacity in striatum of healthy volunteers and performance of cognitive tasks linked to the prefrontal cortex. PMID- 17133404 TI - Folic acid-containing supplement consumption during pregnancy and risk for oral clefts: a meta-analysis. AB - BACKGROUND: There is equivocal evidence in the published literature that folic acid supplementation during pregnancy may protect against the common congenital anomalies cleft lip with or without cleft palate (CLP) and cleft palate alone (CP). We undertook this meta-analysis to test the hypothesis that nonsyndromic oral cleft birth prevalences are different for those whose mothers took folic acid-containing supplements and for those whose mothers did not. METHODS: Human studies published in English were identified through MEDLINE, bibliography reviews, and contacting experts in the field. Within strata of prospective and case-control studies, CLP, CP, and all clefts, respectively, were analyzed using either a fixed or random effects model, as appropriate. We assessed for publication bias using Begg and Mazumdar's rank correlation and Egger's regression-based tests. RESULTS: Five prospective studies were analyzed, yielding combined relative risks of 0.51 (95% CI: 0.32, 0.95) for CLP, 1.19 (95% CI: 0.43, 3.28) for CP, and 0.55 (95% CI: 0.32, 0.95) for all clefts. Twelve case-control studies were assessed, which resulted in combined relative risks of 0.77 (95% CI: 0.65, 0.90) for CLP, 0.80 (95% CI: 0.69, 0.93) for CP, and 0.78 (95% CI: 0.71, 0.85) for all clefts. CONCLUSIONS: In aggregate, our results support the hypothesis of a protective effect of folic acid-containing supplement intake during pregnancy on the risk for oral clefts, although this conclusion is tempered by the potential for bias and uncontrolled confounding. PMID- 17133405 TI - Update of the decitabine experience in higher risk myelodysplastic syndrome and analysis of prognostic factors associated with outcome. AB - BACKGROUND: Therapy for patients with myelodysplastic syndrome (MDS) with hypomethylating agents, like decitabine and 5-azacitidine, has produced favorable results. In this study, the authors update their experience with decitabine in patients with MDS and analyze the cytogenetic response patterns and prognostic factors associated with decitabine therapy. METHODS: One hundred fifteen patients with higher risk MDS who received treatment with decitabine were reviewed. Patients received decitabine 100 mg/m(2) per course every 4 weeks in 3 different schedules: 1) 20 mg/m(2) intravenously daily x 5, 2) 20 mg/m(2) subcutaneously daily x 5, and 3) 10 mg/m(2) intravenously daily x 10. Decitabine was given for a median of >or=7 courses (range, 1-23 courses). RESULTS: Overall, 80 patients (70%) achieved a response according to the modified International Working Group criteria (IWG): complete response (CR), 40 patients (35%); partial response, 2 patients (2%); bone marrow CR with or without other hematologic improvements (HI), 26 patients (23%); and other HI, 12 patients (10%). Cytopenias were improved in 50% of patients. The median remission duration was 20 months, and the median survival was 22 months. Mortality was 3% at 6 weeks and 7% at 3 months. In a multivariate analysis, poor prognostic factors for achieving IWG CR were MDS (vs chronic myelomonocytic leukemia), longer duration of MDS, and prior MDS therapy. For survival, independent adverse prognostic factors were chromosome 5 and/or 7 abnormalities, older age, and prior MDS therapy (excluding growth factors). CONCLUSIONS: The longer term experience with decitabine in MDS was favorable. Pretreatment prognostic factors may predict the outcome of patients who receive decitabine therapy for MDS. PMID- 17133406 TI - How do oncologists deal with incidental abnormalities on whole-body fluorine-18 fluorodeoxyglucose PET/CT? AB - BACKGROUND: Combined positron emission tomography (PET)/computed tomography (CT) using fluorine-18 fluorodeoxyglucose (FDG) is an exciting technique for cancer evaluation, but false-positive results are a recognized limitation. The aim of the study was to evaluate how oncologists deal with focal extrathyroidal FDG abnormalities considered by imaging specialists to be unrelated to the referral indication. METHODS: PET scan reports from a 12-month period from August 2002 to July 2003 in 1727 consecutive patients (mean age, 63 years) were reviewed. Incidental, nonphysiologic FDG abnormalities were classified based on the report conclusion. The frequency with which such abnormalities were investigated by oncologists and the final diagnosis were compared with the imaging diagnosis with a minimum potential follow-up of 2 years (mean, 27.5 months). RESULTS: Incidental FDG abnormalities were reported in 199 (12%) of 1727 patients, including 181 with adequate follow-up. Of 59 cases with a suspected second malignancy, 34 (58%) were actively investigated, with 14 confirmed, 7 unexpected metastatic sites, and 10 other active pathologies. Only 1 further cancer was subsequently detected in the 25 (42%) patients not actively investigated. Conversely, of 122 sites presumed to be benign, only 10 (8%) were actively investigated. Only 2 were proven to relate to malignancy. CONCLUSIONS: Although incidental abnormalities were common, most were benign and appropriately categorized by experienced readers. For actively investigated extrathyroidal abnormalities, a neoplastic basis was confirmed in over 60% of cases. Conversely, for cases deemed most likely benign by the PET/CT report or after review of readily available clinical information by the referring oncologist, the rate of malignancy was less than 2%. PMID- 17133407 TI - Prognostic factors and outcome of recurrence in childhood acute myeloid leukemia. AB - BACKGROUND: Outcome after recurrence of childhood acute myeloid leukemia (AML) is poor. We performed this study to identify prognostic factors for recurrence and for survival after recurrence of AML. METHODS: The clinical characteristics, biological features, treatment modalities, and outcomes of children with de novo AML who were enrolled on 3 consecutive clinical protocols from 1987 to 2002 at St. Jude Children's Research Hospital were studied. Regression modeling was used to identify prognostic factors for recurrence and for survival after recurrence. RESULTS: The outcome after recurrence was poor, with a 5-year survival estimate of only 23.3% +/- 5.7%. Multivariable analysis indicated that male sex (P = .005), autologous stem cell transplant before recurrence (P = .097), each additional month from diagnosis to recurrence (P = .041), and stem cell transplant after recurrence (P < .001) were associated with a better survival after recurrence, whereas M5 or M7 morphology (P = .001) were significantly predictive of a lower survival estimate after recurrence. CONCLUSIONS: Survival after recurrence was poor in children with AML. Novel therapies are urgently needed to prevent or to treat recurring AML. PMID- 17133410 TI - User-friendly semiautomated assembly of accurate image mosaics in microscopy. AB - We present a semiautomated software solution to the problem of extending the lateral field of view of a classical microscope. The initial requirements are a set of overlapping images, along with their user-provided coarse mosaic. Our solution then refines this initial mosaic in a fully automatic fashion. We rely on a highly accurate registration engine to perform the pairwise registration of the individual images, and on an efficient strategy to minimize the amount of computations while maintaining the highest possible global quality. We describe these ingredients, which we make available as a free multiplatform user-friendly software package. We also highlight why and how the specific aspects of the present microscopy application differ from those encountered while creating more common mosaics such as panoramas. Finally, we present experimental results that illustrate and validate our method on a real biological sample. We conclude by showing that we are able to reach subpixel accuracy. PMID- 17133411 TI - Generation of conditional Cited2 null alleles. AB - Cited2 is a transcriptional co-factor that is widely expressed in both embryonic and extraembryonic cells during early development. It is essential for embryonic development with Cited2 null embryos showing abnormal development of organs including heart, neural tube, adrenal glands, and placenta (both in trophoblast derivatives and invading fetal vasculature), as well as having defects in the establishment of the left-right body axis. We report the generation of two conditional null alleles allowing Cre-recombinase-mediated somatic cell gene inactivation. Mice heterozygous or homozygous for these alleles are viable and fertile. Crossing conditional mutants with CMV-Cre transgenic mice produces an embryonic-lethal phenotype in the offspring indistinguishable from germline null mutants. We also demonstrate that conditional deletion results in lacZ expression under the control of the Cited2 promoter. These alleles are therefore useful genetic tools for dissecting the functions of Cited2 in the formation of different organs and patterning of the developing embryo. genesis PMID- 17133412 TI - Experience with heat shock protein-peptide complex 96 vaccine therapy in patients with indolent non-Hodgkin lymphoma. AB - BACKGROUND: The objective of this phase II trial was to investigate the safety and efficacy of autologous heat shock protein-peptide complex 96 (HSPPC-96) vaccines prepared from tumor specimens of patients with newly diagnosed or previously treated indolent non-Hodgkin lymphoma (NHL). METHODS: The study was for patients with indolent B-cell NHL with measurable lesions. HSPPC-96 vaccines were prepared from patients' resected tumor specimens and administered as a 25 microg intradermal injection in the absence of disease progression every week for 4 weeks and then every 2 weeks until the vaccine supply was exhausted. RESULTS: Twenty patients were enrolled in this trial. The median patient age was 59 years. Ten patients had been treated previously (median, 2 regimens; range, 1 to 7). Eighteen (90%) patients had stage III or IV disease. Autologous vaccines were successfully prepared for 17 (85%) patients and all received at least 1 dose. The treatment was very well tolerated. One patient experienced a response with biopsy proven clearance of the lymphoma cells in 2 of the skin nodules at 3.0 months that lasted for 7.0 months. Eight patients had stable disease for 6.0 to 19.8 months. The median failure-free survival duration in patients who received vaccine therapy was 5.2 months. CONCLUSIONS: HSPPC-96 can be prepared from tumor specimens for the majority of lymphoma patients, but it had limited efficacy in inducing responses in patients with active diseases. Further studies of HSPPCs, therefore, should be considered in adjuvant settings or in combination with other immunomodulatory agents to assess survival benefit. PMID- 17133413 TI - Clear cell sarcoma of tendons and aponeuroses: a study of 75 patients. AB - BACKGROUND: Clear cell sarcoma (CCS) of tendons and aponeuroses (malignant melanoma of soft parts) is a rare melanocytic soft tissue sarcoma. The objective of this study was to determine the clinical features, prognostic factors, and optimal treatment policy for patients with this rare disease. METHODS: Seventy five consecutive patients with histologically confirmed CCS who received treatment between 1980 and 2004 were analyzed retrospectively. RESULTS: There were 41 men and 34 women, and the median age was 36 years. Sixty-five tumors were located in the extremities, and 10 tumors were located in the trunk. The median tumor size was 4 cm. Seventy-one patients underwent surgical excision, and 56 patients received chemotherapy. Sixteen patients developed local recurrences, and 52 patients developed metastasis. The overall patient survival rates was 47% at 5 years and 36% at 10 years. Univariate analysis showed that sex (P = .018), tumor size (P = .001), tumor depth (P = .002), TNM classification (P = .001), and surgical margin (P = .042) were significant prognostic factors. Among the 52 patients who presented with localized disease, sex (P = .023), tumor size (P = .002), tumor depth (P = .011), TNM classification (P = .004), and chemotherapy (P = .032) were identified as significant prognostic factors. Multivariate analysis showed that tumor size remained an independent prognostic factor in both groups. CONCLUSIONS: The current results supported the contention that early diagnosis and initial wide excision are essential for a favorable outcome of CCS. The role of chemotherapy for CCS should be investigated further. PMID- 17133414 TI - Targeted gene expression in the zebrafish prechordal plate. AB - Targeted gene expression is a powerful tool for understanding gene function in vivo. In zebrafish, overexpression of gene products is typically accomplished ubiquitously, without temporal and spatial specificity. However, the yeast Gal4/UAS system can be used for targeted gene expression in zebrafish. Here we describe the generation and characterization of Tg[gsc: Gal4-VP16] transgenic zebrafish lines that harbor a construct encoding Gal4-VP16 transcriptional activator under the control of a fragment of the goosecoid gene promoter. Tg[gsc:Gal4-VP16] embryos express Gal4-VP16 RNA in presumptive prechordal plate mesendoderm during late blastula and throughout gastrulation. By crossing these fish to Tg[UAS-GFP] transgenic fish, we show that the gsc:Gal4-VP16 transgene is capable of driving strong expression of a target gene in the prechordal plate and its derivatives during gastrulation and segmentation. Thus, the use of Tg[gsc:Gal4-VP16] fish can help in understanding gene function in the prechordal plate, an embryonic structure that is crucial for normal neural patterning. PMID- 17133415 TI - Extended safety and efficacy data on S-1 plus cisplatin in patients with untreated, advanced gastric carcinoma in a multicenter phase II study. AB - BACKGROUND: S-1 is a promising oral fluoropyrimidine. The authors obtained extended Phase II safety and efficacy data in a multicenter setting for the S-1 plus cisplatin combination: The experimental arm of the global Phase III First Line Advanced Gastric Cancer Study (FLAGS) is being compared with 5 fluorouracil/cisplatin. METHODS: Eligible patients had untreated, histologically confirmed advanced gastric cancer (AGC), a Karnofsky performance status (KPS) > or =70%, adequate organ function, and provided written consent. Patients received S-1 (25 mg/m(2) twice daily on Days 1 through 21) plus cisplatin (75 mg/m(2) on Day 1) every 28 days. The confirmed overall response rate (CORR) also was designated by an external review. The time to progression (TTP), median survival (MS), and safety were assessed. RESULTS: All 72 patients were assessed for safety and survival, and 64 patients were assessed for CORR. The median KPS was 90%. The median number of treatment cycles was 4. The CORR was 55% (95% confidence interval [95% CI], 42-67%). The median duration of response was >5 months. At 6 months, only an estimated 38% of patients had cancer progression. The estimated MS was 10.4 months (95% CI, 8.6-12.9 months). At least 1 serious adverse event occurred in 44% of patients. The frequent grade 3 or 4 toxicities (using the National Cancer Institute Common Toxicity Criteria), which occurred in >10% of patients, included fatigue/asthenia (24%), emesis (17%), nausea (15%), diarrhea (13%), and neutropenia (19%). Complicated neutropenia (1.4%) and grade 4 diarrhea (1.4%) were rare. CONCLUSIONS: The current extended data confirmed that S-1 combined with cisplatin had a highly desirable safety profile. The efficacy against AGC, according to an external review, was encouraging. FLAGS is expected to complete its accrual of 1050 patients by December 2007. PMID- 17133416 TI - Prostate-specific antigen nadir within 12 months of prostate cancer radiotherapy predicts metastasis and death. AB - BACKGROUND: The nadir prostate-specific antigen (PSA) at 1 year (nPSA12) was investigated as an early estimate of biochemical and clinical outcome after radiotherapy (RT) alone for localized prostate cancer.METHODS.From May 1989 to November 1999, 1000 men received 3D conformal RT alone (median, 76 Gy) with minimum and median follow-up periods of 26 and 58 months, respectively, from the end of treatment. The calculation of PSA doubling time (PSADT) was possible in 657 patients. Multivariate analyses (MVAs) via Cox proportional hazards regression were used to determine the association of nPSA12 to biochemical failure (BF; ASTRO definition), distant metastasis (DM), cause-specific mortality (CSM), and overall mortality (OM). Dichotomization of nPSA12 was optimized by evaluating the sequential model likelihood ratio and P-values.RESULTS.In MVA, nPSA12 as a continuous variable was independent of RT dose, T-stage, Gleason score, pretreatment initial PSA, age, and PSADT in predicting for BF, DM, CSM, and OM. Dichotomized nPSA12 (2 versus >2 ng/mL) was independently related to DM and CSM. Kaplan-Meier 10-year DM rates for nPSA12 2 versus >2 ng/mL were 4% versus 19% (P<.0001).CONCLUSIONS.nPSA12 is a strong independent predictor of outcome after RT alone for prostate cancer and should be useful in identifying patients at high risk for progression to metastasis and death. PMID- 17133417 TI - Differences in health and cultural beliefs by stage of mammography screening adoption in African American women. AB - Behavioral studies show that women's stage of readiness to adopt mammography screening affects their screening rates and that beliefs about breast cancer and screening affect stages of screening. The purposes of this study were to determine, first, the relationship between particular health and cultural beliefs and stage of mammography screening adoption in urban African American women, and second, whether demographic and experiential characteristics differed by stage. Data were analyzed from 344 low-income African American women nonadherent to mammography screening who participated in a 21-month trial to increase screening. At baseline, these women were randomized into 1 of 3 groups: tailored interactive computer instruction, targeted video, or usual care. Participants were categorized by stage of mammography screening adoption at 6 months as precontemplators (not planning to have a mammogram), contemplators (planning to have a mammogram), or actors (had received a mammogram). Although demographic and experiential variables did not differentiate stages of screening adoption at 6 months postintervention, some health and cultural beliefs were significantly different among groups. Actors were more preventive-health-oriented than precontemplators and had fewer barriers to screening than did contemplators. Precontemplators had more barriers, less self-efficacy, and greater discomfort with the mammography screening environment than did contemplators or actors. These results will be useful, not to change cultural beliefs, but to guide the design of health education messages appropriate to an individual's culture and health belief system. Cancer 2007. (c) 2006 American Cancer Society. PMID- 17133418 TI - PLP overexpression perturbs myelin protein composition and myelination in a mouse model of Pelizaeus-Merzbacher disease. AB - Duplication of PLP1, an X-linked gene encoding the major myelin membrane protein of the human CNS, is the most frequent cause of Pelizaeus-Merzbacher disease (PMD). Transgenic mice with extra copies of the wild type Plp1 gene, a valid model of PMD, also develop a dysmyelinating phenotype dependant on gene dosage. In this study we have examined the effect of increasing Plp1 gene dosage on levels of PLP/DM20 and on other representative myelin proteins. In cultured oligodendrocytes and early myelinating oligodendrocytes in vivo, increased gene dosage leads to elevated levels of PLP/DM20 in the cell body. During myelination, small increases in Plp1 gene dosage (mice hemizygous for the transgene) elevate the level of PLP/DM20 in oligodendrocyte soma but cause only minimal and transient effects on the protein composition and structure of myelin suggesting that cells can regulate the incorporation of proteins into myelin. However, larger increases in dosage (mice homozygous for the transgene) are not well tolerated, leading to hypomyelination and alteration in the cellular distribution of PLP/DM20. A disproportionate amount of PLP/DM20 is retained in the cell soma, probably in autophagic vacuoles and lysosomes whereas the level in myelin is reduced. Increased Plp1 gene dosage affects other myelin proteins, particularly MBP, which is transitorily reduced in hemizygous mice but consistently and markedly lower in homozygotes in both myelin and naive or early myelinating oligodendrocytes. Whether the reduced MBP is implicated in the pathogenesis of dysmyelination is yet to be established. PMID- 17133419 TI - Acquired von Willebrand syndrome: an update. AB - Acquired von Willebrand syndrome (aVWS) is a rare bleeding disorder with laboratory findings similar to those for congenital von Willebrand disease (VWD). However, unlike congenital VWD, it arises in individuals with no personal or family history of bleeding. aVWS occurs in association with a variety of underlying disorders, most frequently in lymphoproliferative disorders, myeloproliferative disorders, and cardiovascular diseases. Through an analysis of the more recent literature data, the pathophysiology and the clinical, laboratory, and therapeutic aspects of this syndrome are concisely reported in this review. PMID- 17133420 TI - Hyperammonemia and encephalopathy in patients with multiple myeloma. PMID- 17133421 TI - Imatinib mesylate responsiveness in aggressive systemic mastocytosis: novel association with a platelet derived growth factor receptor beta mutation. PMID- 17133422 TI - Pathological splenic rupture: a rare complication of chronic myelomonocytic leukemia. AB - Chronic myelomonocytic leukemia (CMML) is an uncommon disorder characterized by monocytosis of the peripheral blood, absence of the Philadelphia chromosome, fewer than 20% blasts, and one or more lineages showing dysplastic features. Splenomegaly is frequently seen and may be massive. A 56-year-old man with stable CMML and moderate splenomegaly presented to the emergency department with generalized abdominal pain and abrupt drop in hemoglobin. Abdominal imaging revealed splenic rupture and emergency splenectomy was undertaken, with complete recovery. Atraumatic rupture of the spleen has rarely been reported as a complication of CMML or other myelodysplastic disorders. This report should alert physicians to consider this diagnosis in patients with CMML and acute abdominal pain. PMID- 17133423 TI - Clearance of the Janus kinase 2 (JAK2) V617F mutation after allogeneic stem cell transplantation in a patient with myelofibrosis with myeloid metaplasia. AB - A patient with myelofibrosis with myeloid metaplasia displaying the V617F mutation of the JAK2 gene was given an allogeneic stem cell transplantation using a reduced-intensity conditioning regimen. The patient engrafted, and as he became a chimera, the expression of the V617F mutation of the JAK2 gene decreased progressively until its disappearance. Accordingly, the concept of "molecular remission" of the myelofibrosis with myeloid metaplasia could be entertained and added to the categories of response to treatment which have been recently described. PMID- 17133424 TI - Variable adhesion of different red blood cell products to activated vascular endothelium under flow conditions. AB - Red blood cells (RBCs) that have been stored prior to transfusion show increased adherence to vascular endothelium in vitro, which suggests a potential for stored blood transfusion to impede blood flow in some patients. Transfusion is often required in patients with sepsis or inflammation; however, whether activation of endothelium affects stored RBC-endothelial cell (EC) interactions is unknown. We investigated whether storage time and leukocyte content of RBC products influences the adhesion of RBCs to activated ECs. RBCs from nonleukocyte-reduced (S-RBCs), buffy-coat-poor (BCP-RBCs), and leukocyte-filtered (LF-RBCs) products and cultured EC layers were pretreated with endotoxin, tumor necrosis factor alpha (TNF-alpha), or medium alone prior to perfusion of the RBCs across the EC layer in a continuous flow microchamber. After a single day of RBC storage, the number of adherent RBCs was increased in the endotoxin and TNF-alpha pretreated groups compared to the unactivated-control group. These differences were statistically significant for S-RBCs and LF-RBC products (P < 0.05). In contrast, there was no significant difference in RBC adherence to activated and unactivated endothelium at other time-points of RBC product storage. The strength of adhesion of stored RBCs from S-RBC products to activated ECs was not altered following treatment; however, endotoxin significantly increased the adhesive strength of LF RBCs to endothelium. These results demonstrate that while fresh RBCs show increased adhesion to activated endothelium, storage of RBCs did not promote increased adhesion to activated endothelium. However, inflammatory conditions promote stronger adhesion of stored RBCs to ECs, which may contribute to impaired tissue perfusion in some transfusion recipients. PMID- 17133425 TI - Chronic myeloid leukemia following radiotherapy for carcinoma of the cervix: report of a case and brief review of the literature. PMID- 17133426 TI - The high incidence of varicella herpes zoster with the use of bortezomib in 10 patients. AB - Bortezomib, a proteasome inhibitor, has been used for patients with refractory and relapsed multiple myeloma, lymphoma and leukemia. We used bortezomib in ten refractory or relapsed patients (seven of multiple myeloma, two of lymphoma and one of acute myeloblastic leukemia). Six out of ten (60%) patients developed varicella herpes zoster after the complete of one cycle of bortezomib. The incidence of varicella herpes zoster was higher than reported in the literature. It may be due to immunosuppression caused by the combination of high-dose dexamethasone or other drugs. We considered that prophylactic antiviral medication could be used in predisposed patients to reduce the incidence of varicella herpes zoster. PMID- 17133427 TI - Abdominal pain and syndrome of inappropriate antidiuretic hormone secretion as a manifestation of visceral varicella zoster virus infection in a patient with non Hodgkin's lymphoma. PMID- 17133428 TI - The study of sequence configuration and functional impact of the (AC)n(AT)xTy motif in human beta-globin gene promoter. AB - In this report we examine the (AC)n(AT)xTy motif residing -530 bp 5' upstream of the beta-globin gene in Chinese thalassaemic patients. This motif is a putative binding site for a repressor protein, termed beta protein 1 (BP1) (Berg et al., Nucleic Acids Res 1989;17:8833-8852). Variations in the (AC)n(AT)xTy repeats affect the binding affinity of BP1, thereby altering the expression of the beta globin gene. Eight different configurations of this repeat motif are identified in our population of Chinese beta-thalassaemia patients. A (AC)3(AT)7T5 motif was identified among these thalassaemia patients and its influence in beta-globin gene expression was studied using stable transfection assay in murine erythroleukemia (MEL) cells. Our data demonstrated that the (AC)3(AT)7T5 motif has a moderately strong repressor effect on the expression of the cis-linked beta globin gene. The high affinity of BP1 for this motif may result in the suppression of the transcription of the beta-globin gene (Berg et al., Am J Hematol 1991;36:42-47). We postulate that silencer elements in the beta-globin promoter play an important role in modifying the clinical presentation of the disease. PMID- 17133429 TI - T-cell receptor gamma and delta gene rearrangements and junctional region characteristics in south Indian patients with T-cell acute lymphoblastic leukemia. AB - Clonal T-cell receptor (TCR) gamma and delta gene rearrangements were studied in 40 T-ALL cases (pediatrics, 29; adults, 11) using PCR with homo-heteroduplex analysis. At least one clonal TCRG or TCRD rearrangement was detected in 34 (85%) cases. TCR gamma (TCRG) rearrangement was detected in 25 (62.5%) cases that included 16 (55%) pediatrics and 9 (81.8%) adults. TCR delta (TCRD) rearrangement was detected in 14/40 (35%) cases, which included 12 (41%) pediatrics and 2 (18%) adults. The frequency of VgammaI-Jgamma1.3/2.3 was significantly more in adults than pediatrics (81.8% vs. 41.3%, P=0.02). In TCRD, Vdelta1-Jdelta1 was rearranged in 10 (25%) cases. The surface membrane CD3 positive cases are significantly associated with absence of TCRD rearrangements (surface membrane CD3+ TCRdelta- 84% vs. surface membrane CD3- TCRdelta- 48%, P value=0.03). Junctional region sequence analyzed with 10 cases each, of TCRG and TCRD, revealed an average junctional region of 7.4 nucleotides (range 2-18 nucleotides) in TCRG and 27 nucleotides (range 14-42 nucleotides) in TCRD-complete rearrangements. In TCRG, trimming at the ends of Vgamma and Jgamma germline nucleotides resulted in deletion, on an average of 9.2 nucleotides. In TCRD, deletion of nucleotides of the Vdelta and Jdelta gene segments on an average was 3.5 nucleotides. The junctional region of TCRD is more diverse than TCRG; nevertheless, the frequency of TCRG was more than that of TCRD and hence we rely more on TCRG clonal markers to quantitate the minimal residual disease in T-ALL. PMID- 17133430 TI - Fitness and extra-group reproduction in male Verreaux's sifaka: An analysis of reproductive success from 1989-1999. AB - Adult males in social groups often compete with other male group members for access to adult females. In some primate species, males also seek mating opportunities in neighboring social groups. Such extra-group fertilizations (EGFs) provide an additional source of variation in male fitness. This additional component of fitness provided by EGFs must be incorporated into analyses that investigate sources of variation in male lifetime reproductive success. In this study, a model is analyzed in which male fitness over a 10-year sample period is decomposed into additive and multiplicative variance and covariance components. The data come from an ongoing study of a wild population of Verreaux's sifaka (Propithecus verreauxi verreauxi) located at Beza Mahafaly Special Reserve, Southwest Madagascar. Paternity and demographic data for 134 males are used to decompose male fitness into the following three multiplicative components: reproductive lifespan during sample period, fertility, and offspring survival. These multiplicative components are estimated for males reproducing within their resident groups plus (i.e., the additive portion) for males reproducing in neighboring social groups. The analysis shows that variation in fertility makes the largest contribution to variation in total fitness, followed by variation in amount of time spent in sample period (which is a proxy of total reproductive lifespan) and variation in offspring survival. EGFs contribute an important source of variation to male fitness, and numerous factors enhance the opportunities for EGFs in male sifaka. These include female choice, a high degree of home range overlap, and a limited mating season. PMID- 17133431 TI - Biodistance analysis of the Moche sacrificial victims from Huaca de la Luna plaza 3C: Matrix method test of their origins. AB - The purpose of this study is to test two competing models regarding the origins of Early Intermediate Period (AD 200-750) sacrificial victims from the Huacas de Moche site using the matrix correlation method. The first model posits the sacrificial victims represent local elites who lost competitions in ritual battles with one another, while the other model suggests the victims were nonlocal warriors captured during warfare with nearby polities. We estimate biodistances for sacrificial victims from Huaca de la Luna Plaza 3C (AD 300-550) with eight previously reported samples from the north coast of Peru using both the mean measure of divergence (MMD) and Mahalanobis' distance (d2). Hypothetical matrices are developed based upon the assumptions of each of the two competing models regarding the origins of Moche sacrificial victims. When the MMD matrix is compared to the two hypothetical matrices using a partial-Mantel test (Smouse et al.: Syst Zool 35 (1986) 627-632), the ritual combat model (i.e. local origins) has a low and nonsignificant correlation (r = 0.134, P = 0.163), while the nonlocal origins model is highly correlated and significant (r = 0.688, P = 0.001). Comparisons of the d2 results and the two hypothetical matrices also produced low and nonsignificant correlation for the ritual combat model (r = 0.210, P = 0.212), while producing a higher and statistically significant result with the nonlocal origins model (r = 0.676, P = 0.002). We suggest that the Moche sacrificial victims represent nonlocal warriors captured in territorial combat with nearby competing polities. PMID- 17133432 TI - Morphometric estimation of torsional stiffness and strength in primate mandibles. AB - In comparative studies of masticatory function and mandibular biomechanics, the mediolateral dimension of the postcanine corpus (corpus breadth) is commonly utilized as a measure of torsional stiffness from which relative torsional strength is inferred. The use of this dimension entails certain assumptions about corpus shape and cortical bone distribution that are invalid. When corpus breadth is related to an appropriate, empirically supported measure of torsional strength, it is revealed that this dimension has limited utility for inference of biomechanical competence under torsion. The use of linear dimensions to infer structural adaptations to specific loading regimes is problematic given that bone tissue is not optimally deployed to minimize strain levels arising from isolated loads. For the inference of the masticatory biomechanical environment, the more reasonable approach is to consider overall size of the corpus (i.e., cross sectional area) for inference of intra- and inter-specific differences in masticatory forces. PMID- 17133433 TI - Genetic studies in French Guiana populations: Synthesis. AB - Twelve blood group and protein systems from a total of 819 individuals from six tribal groups (Apalai-Wayana, Emerillon, Kalina, Palikur Wayampi, and Wayana) living in French Guiana and Brazil were compared with each other and integrated with previous results from 17 other South Amerindian populations studied for the same genetic markers. Using correspondence analysis, map methodologies, and maximum linkage cluster analysis developed with the UPGMA method, we attempted to establish the genetic position of these tribes among South American Indians. Peripheral positions for the Emerillon and the Palikur were observed. Ethnohistorical data in French Guiana suggest that a strong founder effect for the former and endogamy for the latter could have generated the genetic differentiation of these two ethnic groups. However, when considered in a wider context, all French Guiana Natives cluster together in an intermediate position as compared with 17 other Amerindian groups studied for the comparison. PMID- 17133434 TI - Hormones and body size evolution in papionin primates. AB - This study examines the evolution of size differences among papionin primates by measuring hormones that regulate size growth during ontogeny and influence ultimate adult size (insulin-like growth factor-I (IGF-I), insulin-like growth factor binding protein-3 (IGFBP-3), growth hormone binding protein (GHBP), dehydroepiandrosterone sulfate (DHEAS), testosterone, estradiol). The analyses assess longstanding ideas about circulating hormone levels and body size. Importantly, because the consensus papionin molecular phylogeny implies at least two episodes of size increase, this study offers opportunities to determine whether or not similar hormone profiles regulate this apparent evolutionary convergence (i.e., do larger-bodied papionins have higher levels of growth related hormones than smaller-bodied papionins?). Five hundred and sixty serum samples (from 161 individuals) from 11 papionin species were analyzed using a two level approach to address this issue. One used mixed longitudinal samples from two papionin species to test whether, during growth, large- and small-bodied species have higher and lower hormone levels, respectively. The second compared multiple papionin species to assess whether or not hormone levels covary with size in adult animals. Result show that size and hormone levels do not covary consistently across papionins, either during growth or in adulthood. Specifically, some smaller-bodied papionin species have higher absolute hormone levels than larger-bodied species. Differences in some hormone levels appear to track phylogeny more closely than body size. In contrast to studies based on single species, we demonstrate that, while the hormones analyzed affect growth, absolute circulating hormone levels either during growth or adulthood may be decoupled from interspecific differences in body size. PMID- 17133435 TI - Effect of chewing betel nut on measurements of salivary progesterone and estradiol. AB - The measurement of steroids in saliva is both simple and non-invasive and has been widely used in field and clinical-based research. The observance of particular cultural practices by some populations, however, may hamper accurate hormonal analyses. The present study evaluated the effects of one such practice the chewing of betel nut-on the accurate measurement of salivary progesterone and estradiol. A time series experiment was conducted among Bangladeshi women who are regular users of betel nut. Salivary steroids were analyzed by radioimmunoassay in samples collected prior to and then 30, 60, 120, and 240 min following betel quid use. Results show no significant difference between basal steroid levels and those obtained 60, 120, and 240 min after chewing betel nut. We conclude that with specific collection protocols that take into account time since chewing, salivary steroid analyses can be undertaken in populations among whom the practice of chewing betel nut is endemic. PMID- 17133436 TI - On the time scale of New World primate diversification. AB - New World primates comprise a diverse group of neotropical mammals that suddenly appeared in the Late Oligocene deposits of South America at around 26 million years ago (MYA). Platyrrhines seem to have separated from Old World anthropoids ca. 35 MYA, and their subsequent diversfication is not well documented in the fossil record. Therefore, molecular clock studies were conducted to unveil the temporal scenario for the evolution of the group. In this study, divergence times of all splits within platyrrhines until the generic level were investigated, using two different gene data sets under relaxed molecular clocks. Special attention was paid to the basal diversification of living platyrrhines and to the basal split of the modern Cebidae family, since these nodes were reported to be phylogenetically difficult to resolve. The results showed that analyses from various genomic regions are similar to estimates obtained by early single-gene studies. Living New World primates are descendants of ancestors that lived in the Early Miocene, at around 20 MYA, and modern Cebidae and Pitheciidae appeared ca. 16.9 and 15.6 MYA, respectively. The last common ancestor of living Atelidae is 12.4 million years old, making this clade the youngest New World primate family; at approximately the same time, modern Callitrichinae was evolving (11.8 MYA). The gap between the Platyrrhini/Catarrhini separation and the last common ancestor of living Platyrrhini may be as big as 20 million years. Paleontological and geoclimatological evidence corroborates that the sudden appearance of modern families may be a consequence of environmental changes during the Miocene. PMID- 17133437 TI - Demographic and evolutionary trajectories of the Guarani and Kaingang natives of Brazil. AB - A total of 278 individuals from two Brazilian Indian tribes (Guarani and Kaingang) living in five different localities had their mitochondrial DNA sequenced for the first hypervariable segment (HVS-I), and a fraction of them was also studied for seven biallelic Y-chromosome polymorphisms. Nineteen HVS-I lineages were detected, which showed distinct distributions in the two tribes. The G(ST) value obtained with the mtDNA data is about 5 times higher for the Guarani as compared to the Kaingang, suggesting a higher level of differentiation between the three Guarani partialities than between the two Kaingang villages. Non-Amerindian admixture varied with sex and in the Guarani was only observed through the paternal line. Using these data and those of other Tupian and Jean tribes, it was possible to make inferences about past migratory movements and the genetic differentiation of these populations. PMID- 17133438 TI - No signature of Y chromosomal resemblance between possible descendants of the Cimbri in Denmark and Northern Italy. AB - Two European populations are believed to be related to the ancient Germanic tribe Cimbri: one living in Northern Italy, the other living in Jutland, Denmark. The people called Cimbri are documented in the ancient Roman historical record. Arriving from the far north their movements can be tracked from successive battles with the Romans. The Cimbri finally entered Italy from the northeast and were defeated at Vercellae (present day Vercelli) in 101 BC by Gaius Marius and his professional legions. Classical sources from the first centuries AD relate the homeland of the Cimbri to the coasts around the Elb estuary (northern Germany) or specifically towards the north (Himmerland in northern Jutland). In the alpine parts of Veneto, northeast of the historical battlefield, local traditions dating back to late medieval time, identify a local population as Cimbri living in Terra dei Cimbri. They are considered the descendents of the Germanic combatants that fled the battlefield at Vercelli. As the defeated Cimbri that possibly fled to the mountains of Northern Italy most likely would have been male (warriors), the present study investigated the possible Y chromosomal diversity of the two present populations using microsatellite markers and single nucleotide polymorphisms. While Cimbri from Himmerland resembled their geographical neighbors from Denmark for the Y-chromosome markers, Cimbri from Italy were significantly differentiated both from Cimbri from Himmerland and from Danes. Therefore, we were not able to show any biological relationship for uniparentally transmitted markers. PMID- 17133439 TI - Changes in regional cerebral blood flow following mood challenge in drug-free, remitted patients with unipolar depression. AB - Studies of mood provocation in remitted patients with depression have enabled the description of specific brain changes relevant to depression relapse vulnerability. Because patients in these studies were also receiving maintenance pharmacotherapy, the ability to subtract out the drug effect from the changes observed is reduced. Our study addresses this concern. Changes in regional cerebral blood flow (rCBF) among 11 medication-free, remitted patients with unipolar depression relative to baseline state were assessed with [(15)O]H(2)O positron emission tomography (PET) following provocation of sadness. Participants showed decreased activation in prefrontal cortex similar to those reported for remitted, medicated patients undergoing mood challenge, as well as for acutely depressed patients who were medication free. Previously reported mediofrontal changes in remitted patients are unlikely to be a consequence of maintenance medication and more likely are evidence of relapse vulnerability. PMID- 17133440 TI - Prognostic factors for survival after stereotactic radiosurgery vary with the number of cerebral metastases. AB - BACKGROUND: Little is understood about the factors that influence survival in patients who undergo gamma-knife stereotactic radiosurgery (SRS) for brain metastases. METHODS: Demographic, disease, treatment, and survival data on 334 patients with intracranial metastases who underwent initial SRS from 1998 to 2004 were abstracted from treatment records and from the Connecticut Tumor Registry. Multivariate survival analysis was used to identify factors that independently affected survival. RESULTS: The median age of the patient population was 57.3 years. The median number of lesions treated in a single session was 2 (range, 1 36 lesions treated). The most common tumor histologies were nonsmall cell lung carcinoma (36%), breast cancer (16%), and melanoma (16%). Three hundred patients (90%) had confirmed deaths; the median survival after SRS was 8.1 months. Survival was significantly better in patients who had from 1 to 3 metastases (median, 8.5 months) compared with patients who had > or =4 metastases (median, 6.3 months; hazard ratio [HR], 0.65; P = .003). In the subgroup of patients who had from 1 to 3 metastases, systemic control (HR, 0.49; P < .001), breast cancer (HR, 0.57; P = .003), and total tumor volume < 5 cc (HR, 0.65; P = .002) were associated independently with increased survival, and esophageal cancer (HR, 2.36; P = .042) was associated with decreased survival. In the subgroup of patients who had > or =4 metastases, only age <45 years was associated independently with increased survival (HR, 0.39; P = .006); and melanoma (HR, 2.32; P = .008) and the receipt chemotherapy (HR, 2.59; P = .077) were associated with decreased survival. Sex, race, metastases location, whole-brain radiation, and cranial surgery had no independent associations with altered survival. CONCLUSIONS: The data from this study suggested that different factors affected survival in patients who had from 1 to 3 metastases and patients who had > or =4 metastases. Further research into this area may clarify causes for this discrepancy and improve prognostication. PMID- 17133441 TI - Correlations of quality of life with adverse effects of medication, social support, course of illness, psychopathology, and demographic characteristics in patients with panic disorder. AB - Care of patients with panic disorder has raised quality-of-life (QOL) issues. The purpose of this study was to identify the level of QOL in patients with panic disorder and to examine the relationships between QOL and adverse effects of medication, social support, course of illness, psychopathology, and demographic characteristics. We recruited 57 patients with panic disorder from outpatient psychiatric clinics. We measured their QOL levels with the Short Form of the World Health Organization Questionnaire on Quality of Life-Taiwan Version (the WHOQOL-BREF Taiwan version) and examined the correlates of QOL. The analysis revealed that multiple factors were associated with poor QOL in patients with panic disorder, including severe adverse effects of medication for panic disorder, perceived low social support, severe current panic symptoms, total Beck Depression Inventory-II (BDI-II) score>/=17, young age, being unmarried, and early onset of panic disorder. The QOL of patients with panic disorder was correlated to multiple factors that were specific to individual subjects and influenced by interactions with treatment and the social environment. The results provide screening factors so that clinicians can intervene to improve QOL for their patients with panic disorder. PMID- 17133442 TI - Lithium therapy for unipolar and bipolar depression among the middle-aged and older adult patient subpopulation. AB - Affective disorders are a major cause of morbidity and disability among middle aged and older people. Thus, the prophylaxis of both unipolar depression and bipolar disorder in this patient subpopulation is an important task of psychiatrists and other physicians. Although lithium remains an effective prophylactic and treatment agent in younger individuals with bipolar disorder, its prophylactic efficacy and tolerability has not been thoroughly investigated among middle aged and older people with unipolar depression and bipolar disorder. Our study is based on a mirror-image design that compared the clinical course with lithium treatment and the clinical course prior to lithium treatment based on a retrospective chart review. We examined the results obtained with long-term lithium maintenance in a group of 60 middle-aged and older adult patients (age >60 years) with unipolar depression and bipolar disorder. More specifically, we analyzed changes of frequency, severity, and duration of depressive or manic relapses, rate and duration of hospitalizations and suicidal behavior (thoughts or attempts), and various assessments of outcome. A significant reduction was found on all indices during lithium therapy compared to before lithium treatment, attesting to the prophylactic efficacy of long-term lithium in unipolar depression and bipolar disorder. The range of side effects in our sample was similar to that found in other reports in this age group. The probability of relapse and recurrence in patients with bipolar disorder and with unipolar depression can be significantly decreased by lithium prophylaxis. Further investigation is mandated to confirm these findings under double-blind conditions. PMID- 17133443 TI - Integrin alpha(5) controls osteoblastic proliferation and differentiation responses to titanium substrates presenting different roughness characteristics in a roughness independent manner. AB - Integrin alpha(5)beta(1) regulates osteoblast proliferation and differentiation on smooth synthetic surfaces presenting different chemistries, but it is not known whether this integrin controls osteoblast behavior on surfaces that have micron-scale rough topographies. We cultured MG63 human osteoblast-like cells on titanium substrates with three different roughness characteristics: chemically polished (PT), grit blasted and acid etched with a complex topography consisting of 20-100 mum craters and 0.5-2 mum micropits (SLA), and plasma-sprayed Ti with irregular projections (TPS). Cells spread well on PT but displayed a smaller footprint on SLA or TPS. Nuclei were larger on PT as well. alpha(5)beta(1) binding and FAK phosphorylation were greater on the rougher surfaces (TPS > SLA > PT). Antibodies against the alpha(5)beta(1) binding site on fibronectin had no effect on cell number at 3 days, but [(3)H]-thymidine incorporation was increased, suggesting that binding to fibronectin was necessary for cell cycle regulation. Antibodies to the alpha(5) subunit reduced cell number at 3 days on PT and TPS and reduced DNA synthesis on all substrates in a surface microstructure-independent manner. At 7 days, cell numbers were reduced on PT, and DNA synthesis was reduced by 50% on all surfaces. At 7 days, anti-alpha(5) antibodies caused a partial reduction in alkaline phosphatase enzyme activity on all surfaces, but this effect was independent of surface microstructure. These results indicate that surface micron-scale topography modulates alpha(5)beta(1) integrin binding and FAK activation. Signaling via alpha(5)-dependent mechanisms is required for DNA synthesis and regulation of alkaline phosphatase, but this effect is independent of surface microstructure. PMID- 17133444 TI - Energy absorption characterization of human enamel using nanoindentation. AB - Enamel is a natural composite, which has much higher toughness than its major component, crystalline hydroxyapatite. In this study, the energy absorption behavior of human sound enamel was investigated with nanoindentation techniques. A UMIS nanoindenter system as well as a Berkovich and two spherical indenters with nominal tip radii of 5 and 20 microm were used to indent enamel at different loading forces in the direction parallel to enamel prisms. Inelastic energy dissipation versus depth of indenter penetration (U%-h(p) curve) as well as a function of indentation strain (U%-epsilon curve) of enamel was determined. Enamel showed much higher energy absorption capacity than a ceramic material with equivalent modulus (fused silica). Even at the lowest forces (1 mN) for the 20 microm indenter, inelastic response was found. Additional tests done at different force loading rates illustrated that load rate has little influence on P-h response of enamel. The top surface of enamel has the plastic work of indentation of approximately 5.2 nJ/microm(3). The energy absorbing ability is influenced by the very small protein rich component that exists between the hydroxyapatite nanocrystals as well as within the sheath structure surrounding the enamel rods. PMID- 17133445 TI - Plasma-controlled nanocrystallinity and phase composition of TiO2: a smart way to enhance biomimetic response. AB - This contribution sheds light on the role of crystal size and phase composition in inducing biomimetic apatite growth on the surface of nanostructured titania films synthesized by reactive magnetron sputtering of Ti targets in Ar+O(2) plasmas. Unlike most existing techniques, this method enables one to deposit highly crystalline titania films with a wide range of phase composition and nanocrystal size, without any substrate heating or postannealing. Moreover, by using this dry plasma-based method one can avoid surface hydroxylation at the deposition stage, almost inevitable in wet chemical processes. Results of this work show that high phase purity and optimum crystal size appear to be the essential requirement for efficient apatite formation on magnetron plasma fabricated bioactive titania coatings. PMID- 17133446 TI - Quantification of fibrinogen adsorption onto 316L stainless steel. AB - Adsorption of the plasma protein fibrinogen (Fb) onto 316L stainless steel (316L SS) was observed and quantified using both in situ and ex situ atomic force microscopy techniques. Industry standard mechanical and electrochemical polishing techniques were used to prepare bulk alloy 316L SS samples, rendering the surfaces flat enough to directly observe and measure Fb adsorption. The data were analyzed kinetically using a Langmuir model. Largely irreversible adsorption was found on the 316L SS surface with an adsorption rate constant (k(o)) of 1.9 x 10( 4) mL microg(-1) s(-1) using the ex situ method and 1.7 x 10(-4) mL microg(-1) s( 1) using the in situ method. Additionally, protein conformation and assembly orientation on these surfaces were documented, where the adsorption pattern appeared random. Complete area coverage was never obtained. That is, after adsorption for over 5 time constants (5tau), voids in the structure were always observed. PMID- 17133447 TI - Composites of poly(lactide-co-glycolide) and the surface modified carbonated hydroxyapatite nanoparticles. AB - To improve the mechanical properties of the composites of poly(lactide-co glycolide) (PLGA, LA/GA = 80/20) and the carbonate hydroxyapatite (CHAP) particles, the rice-form or claviform CHAP particles with 30-40 nm in diameter and 100-200 nm in length were prepared by precipitation method. The uncalcined CHAP particles have a coarse surface with a lot of global protuberances, which could be in favor of the interaction of the matrix polymer to the CHAP particles. The nanocomposites of PLGA and surface grafted CHAP particles (g-CHAP) were prepared by solution mixing method. The structure and properties of the composites were subsequently investigated by the emission scanning electron microscopy, the tensile strength testing, and the cell culture. When the contents of g-CHAP were in the range of 2-15 wt %, the PLGA/g-CHAP nanocomposites exhibited an improved elongation at break and tensile strength. At the 2 wt % content of g-CHAP, the fracture strain was increased to 20% from 4-5% for neat PLGA samples. Especially at g-CHAP content of 15 wt %, the tensile strength of PLGA/g-CHAP composite was about 20% higher than that of neat PLGA materials. The tensile moduli of composites were increased with the increasing of filler contents, so that the g-CHAP particles had both reinforcing and toughening effects on the PLGA composites. The results of biocompatibility test showed that the higher g-CHAP contents in PLGA composite facilitated the adhesion and proliferation properties of osteoblasts on the PLGA/g-CHAP composite film. PMID- 17133448 TI - Influence of gradual introduction of hydrophobic groups (stearic acid) in denatured atelocollagen on fibroblasts behavior in vitro. AB - To prepare new biocompatible hydrophobic collagen films for medical devices, innovative collagen derivatives were synthesized by reaction of the lysyl amino groups of the alpha-chains with activated stearic acid. Different collagens having different substitution degrees were obtained and used to prepare films crosslinked with oxidized glycogen. Their physicochemical surface properties were evaluated, and in vitro assays were performed to analyze the behavior of fibroblasts in contact with the materials. The assays were performed with cells in adhesion and growth phases. The hydrophobic properties increased with the number of stearic acid introduced in the collagen but only in the range of 1-12 stearic acids per molecule. For higher modifications a decrease of hydrophoby was observed. All the films induced a decrease of cells growth and adhesion but without cytotoxicity. These effects were more pronounced for the collagen containing about eight stearic acid residues. Cells behavior on modified collagens films seems to be related to the chemical groups exposed on the surface of the films. Indeed, the surface chemistry directly influences the adsorption of adhesion proteins and modulates their conformation therefore modifying the cell adhesion. PMID- 17133449 TI - Electrochemical characterization of albumin protein on Ti-6AL-4V alloy immersed in a simulated plasma solution. AB - The effect of oxygen and albumin on the electrochemical behavior of a Ti-6Al-4V alloy immersed in a simulated inorganic plasma (SIP) solution was studied with a rotating-cylindrical electrode configuration to focus on the surface/electrolyte reactions. Potentiokinetic scans and electrochemical impedance spectroscopy have been used to characterize the interface by determining the passive current density and capacitance. For the polarization scans, an albumin addition of 37.7 mg/cm(3) to the SIP solution (oxygenated and unoxygenated) decreased the passive current density, indicating a lowering of the corrosive rate. The surface capacitance for the Ti-6Al-4V alloy immersed in a SIP solution averaged 13 microF/cm(2), which transformed after albumin addition (37.7 mg/cm(3)) from a potential independent behavior to the capacitance ranging from 23 to 6 microF/cm(2) with increasing potentials from -800 to 1500 mV(SCE), respectively, indicative of albumin adsorption. Within the same potential range and albumin addition to oxygenated solutions, the capacitances expanded slightly with a similar decreasing trend from 31 to 6 microF/cm(2), although the capacitance depicts an interaction between the hydrated passive film and the adsorbed albumin from -550 to 500 mV(SCE) in which the capacitance plateaued at 15 microF/cm(2). The hydrated porous oxide film results from the porous rutile layer reacting with H(2)O(2) formed as an intermediary component of oxygen reduction at the Ti-6Al-4V surface. The passive film-albumin interaction would affect the processing of titanium alloys in their surface preparation for biocompatibility, as well as determining the reactivity of titanium alloys to proteins. PMID- 17133450 TI - Influence of the activator in an acrylic bone cement on an array of cement properties. AB - In all but one of the acrylic bone cement brands used in cemented arthroplasties, N,N-dimethyl-4-toluidine (DMPT) serves as the activator of the polymerization reaction. However, many concerns have been raised about this activator, all related to its toxicity. Thus, various workers have assessed a number of alternative activators, with two examples being N,N-dimethylamino-4-benzyl laurate (DMAL) and N,N-dimethylamino-4-benzyl oleate (DMAO). The results of limited characterization of cements that contain DMAL or DMAO have been reported in the literature. The present work is a comprehensive comparison of cements that contain one of these three activators, in which the values of a large array of their properties were determined. These properties range from the setting time and maximum exotherm temperature of the curing cement to the variation of the loss elastic modulus of the cured cement with frequency of the applied indenting force in dynamic nanoindentation tests. The present results, taken in conjunction with those presented in previous reports by the present authors and co-workers on other properties of these cements, indicate that both DMAL and DMPT are suitable alternatives to DMPT. PMID- 17133451 TI - Silica-ceramic suppresses the expression of proinflammatory cytokines induced by lipopolysaccharide in macrophages. AB - Bioactive materials have previously been used to coat implants. In a new development for bioactive materials, a silica-ceramic mixture was found to alleviate pain (Lee, Poster presented at the Ninth World Congress of Gynecological Endocrinology, Hongkong, 2001. Poster session (p47)). Here, we hypothesized that silica-ceramic can reduce the expression and activity of cyclooxygenase 2 (COX2) or cytokines associated with inflammation. The production of COX2 and proinflammatory cytokines was investigated by reverse transcriptase (RT)-PCR and ELISA assay in macrophages stimulated by lipopolysaccharide (LPS). Silica-ceramic had no effect of COX2 expression and prostaglandin production in macrophages. However, silica-ceramic suppressed the synthesis of cytokines involved in inflammation, in particular, the expression of IL-1beta and IL-6 was reduced at the transcriptional and translational levels. The involvement of NF kappaB in the suppression of cytokines by silica-ceramic was examined by luciferase reporter assay. The NF-kappaB activity stimulated by LPS was inhibited by 20-60% with silica-ceramic compared with treatment with LPS alone. We suggest that inhibition of NF-kappaB activity by silica-ceramic might cause the attenuation of proinflammatory cytokine expression in macrophages. In conclusion, silica-ceramic could be an alternative approach to regulate the inflammation process. PMID- 17133452 TI - Preparation and preliminary characterization of concentric multi-walled chitosan microspheres. AB - Chitosan was first converted into micro-droplets by using a high voltage electrostatic field system. The droplets were then dropped into a series of Na(5)P(3)O(10)/NaOH solution mixtures with volume ratio of 17:3, 19:1, 1:0 (pure aqueous Na(5)P(3)O(10)) or 0:1 (pure aqueous NaOH) in order to fabricate chitosan microspheres with different membrane structures. The microspheres exhibit distinct chemical and physical properties, including release behaviors of encapsulated drugs. These chitosan microspheres prepared by this method exhibited good sphericity within the range of (286.6 +/- 15.9) to (356 +/- 9.5) microm in diameters. SEM observations have indicated that the chitosan microspheres exhibited distinct surface structures depending on the post-treatment solutions. The mechanical strength of the chitosan microspheres significantly improved upon treatment with Na(5)P(3)O(10)/NaOH solution at ratio of 17:3 (v/v), as compared with the same but at ratio of 19:1, 1:0 (pure Na(5)P(3)O(10)) and 0:1 (pure NaOH) solutions. In addition, chitosan microspheres with unique multi-walled concentric shell membrane structures were prepared by treating with Na(5)P(3)O(10)/NaOH solution at ratio of 19:1. Release studies were carried out to evaluate the kinetic profiles of two model drugs (5-fluorouracil and cytochrome C) from these prepared chitosan microspheres. When chitosan microspheres treated with Na(5)P(3)O(10)/NaOH ratio at 17:3, the release of cytochrome C was found to be the slowest as compared to those treated by the same Na(5)P(3)O(10)/NaOH solution of other mixing ratios, after a period of 35-day "endurance" test. However, in one case, 5-fluorouracil released quite quickly in a period of 30 min (about 80% completion). The wide range of drug release results might be attributed to the unique and wide range of surface characteristics, porosities, and various structures of chitosan microspheres upon treatment with Na(5)P(3)O(10)/NaOH solutions. These results indicate that, by adjusting the Na(5)P(3)O(10)/NaOH ratios, without extra manipulation on polymer material formulation, one could obtain an additional degree of freedom in drug release profile that permits the simultaneous regulation of morphologies of surface texture and internal structure, mechanical properties, and molecular permeability of the microspheres. PMID- 17133453 TI - Mechanical stimulation induces morphological and phenotypic changes in bone marrow-derived progenitor cells within a three-dimensional fibrin matrix. AB - One of the major limitations in tissue engineering is cell sourcing. Multipotent progenitor cells appear to have many promising features for that purpose. Mechanical stimulation is known to play an important role in determining cell phenotype. The aim of this work was to investigate the effects of cyclic stretch on rat bone marrow derived progenitor cell (BMPC) morphology and smooth muscle directed differentiation within a three-dimensional fibrin matrix. BMPCs were suspended in a fibrin gel, pipetted into the trough of Flexcell Tissue-Train plates, and stimulated with 10% longitudinal cyclic stretch at 1 Hz for 6 days. Unconstrained (stress- and strain-free) and static anchored (constrained but not stretched) samples were used as controls. Stress filament area per cell was increased in the stretched samples compared to static anchored and free-float controls. Cells in the free float controls were randomly aligned, while they aligned parallel to the direction of the stress or strain in the other groups. Immunofluorescence suggested an increased expression of smooth muscle markers (smooth muscle alpha actin and h1-calponin) in both stretched and constrained control samples, but not in unconstrained controls. Qualitative assessment suggested that collagen production was increased in both mechanically stimulated samples. Proliferation was inhibited in stretched samples compared to the constrained controls. This work suggests an ability of rat BMPCs to differentiate toward a smooth-muscle-cell-like lineage when exposed to biomechanical stimulation in a three-dimensional model. The observation that the constrained samples induced changes in BMPCs suggests that stress alone may be stimulatory, but addition of cyclic stretch appears to augment the responses. PMID- 17133454 TI - Regional metabolic changes in parkinsonian patients with normal dopaminergic imaging. AB - Dopaminergic imaging has been found to be normal in approximately 15% of parkinsonian patients enrolled in neuroprotective trials. We used (18)F fluorodeoxyglucose positron emission tomography (FDG PET) to determine the metabolic basis for this finding. We reviewed scans from 185 patients with clinical signs of Parkinson's disease (PD) who underwent (18)F-fluorodopa PET imaging for diagnostic confirmation. Of this group, 27 patients (14.6%) had quantitatively normal scans; 8 of these patients were additionally scanned with FDG PET. Pattern analysis was performed on an individual scan basis to determine whether the metabolic changes were consistent with classic PD. Computer-assisted single-case assessments of the FDG PET scans of these 8 patients did not disclose patterns of regional metabolic change compatible with classical PD or an atypical parkinsonian variant. Similarly, network quantification revealed that PD-related pattern expression was not elevated in these patients as it was in an age- and duration-matched cohort with classical PD (P < 0.0001). None of these patients developed clinical signs of classical PD or of an atypical parkinsonian syndrome at a follow-up visit conducted 3 years after imaging. The results suggest that parkinsonian subjects with normal dopaminergic imaging do not have evidence of classical PD or an atypical parkinsonian syndrome. PMID- 17133455 TI - Asynchronous blepharospasm, facial and cervical dystonia, and bilateral asynchronous hemifacial spasm. AB - We present a patient with a facial movement disorder that has characteristics of both blepharospasm and bilateral asynchronous hemifacial spasm. Because of the increased incidence of blepharospasm in patients with hemifacial spasm, our patient's clinical presentation is probably not a chance occurrence, but rather a manifestation of some predisposition for these two movement disorders. This unusual constellation of signs and symptoms challenges the current diagnostic criteria and suggests that some of these facial movement disorders may lie on a spectrum, rather than represent distinct entities. PMID- 17133456 TI - Prediction of recurrence of hepatocellular carcinoma after curative ablation using three tumor markers. AB - Three tumor markers for hepatocellular carcinoma (HCC) are available in daily practice in Japan: alpha-fetoprotein (AFP), des-gamma-carboxy prothrombin (DCP), and lens culinaris agglutinin-reactive fraction of alpha-fetoprotein (AFP-L3). To elucidate the predictability of these tumor markers on HCC recurrence after curative ablation, we enrolled 416 consecutive patients with naive HCC who had been treated by percutaneous ablation at our department from July 1997 to December 2002. Tumor marker levels were determined immediately before and 2 months after the treatment. Complete ablation was defined on CT findings as nonenhancement in the entire lesion with a safety margin. Tumor recurrence was also defined as newly developed lesions on CT that showed hyperattenuation in the arterial phase with washout in the late phase. We assessed the predictability of recurrence via tumor markers in multivariate analysis, using proportional hazard regression after adjusting for other significant factors in univariate analysis. Until the end of follow-up, tumor recurrence was identified in 277 patients. Univariate analysis revealed the following factors to be significant for recurrence: platelet count; size and number of tumors; AFP, AFP-L3, and DCP preablation; and AFP and AFP-L3 postablation. Multivariate analysis indicated that AFP >100 ng/mL and AFP-L3 >15%, both pre- and postablation, were significant predictors. The positivity of AFP and AFP-L3 preablation that turned negative postablation was not significant. In conclusion, tumor markers pre- and post ablation were significant predictors for HCC recurrence and can complement imaging modalities in the evaluation of treatment efficacy. PMID- 17133457 TI - Role of the JAK2 mutation in the diagnosis of chronic myeloproliferative disorders in splanchnic vein thrombosis. AB - The diagnosis of an underlying chronic myeloproliferative disorder (CMPD) is often problematic in patients with primary extrahepatic portal vein obstruction (EHPVO) or Budd-Chiari syndrome (BCS); indeed, conventional clinical and hematological parameters usually yield insufficient information. To assess the diagnostic contribution of the gain-of-function mutation V617F of the JAK2 gene, 93 patients with EHPVO or BCS were investigated. JAK2 V617F was identified in 35.6% of 73 patients with EHPVO and in 40% of 20 patients with BCS. Taking the JAK2 mutation as a test with the highest positive predictive value for the diagnosis of CMPD, conventional clinical-hematological parameters had a sensitivity for CMPD lower than 48%. Bone marrow (BM) histology provided a diagnosis of CMPD in 41/74 (55.4%) patients, with a sensitivity of 93.5%. Clonality of hematopoiesis as assessed by granulocyte X-chromosome inactivation was present in 65.1% of 43 informative female patients, with a sensitivity of 86.6%. By resolving the sensitivity bias of the JAK2 mutation with the results of BM histology and clonality assay, CMPD was diagnosed in 53% of patients with EHPVO or BCS. In conclusion, CMPD is the major cause of primary EHPVO or BCS. JAK2 V617F is a very reliable and noninvasive molecular marker for CMPD and should be used as a first test for diagnosis. PMID- 17133458 TI - Hyponatremia in cirrhosis: Results of a patient population survey. AB - Low serum sodium concentration is an independent predictor of mortality in patients with cirrhosis, but its prevalence and clinical significance is unclear. To evaluate prospectively the prevalence of low serum sodium concentration and the association between serum sodium levels and severity of ascites and complications of cirrhosis, prospective data were collected on 997 consecutive patients from 28 centers in Europe, North and South America, and Asia for a period of 28 days. The prevalence of low serum sodium concentration as defined by a serum sodium concentration < or =135 mmol/L, < or =130 mmol/L, < or =125 mmol/L, and < or =120 mmol/L was 49.4%, 21.6%, 5.7%, and 1.2%, respectively. The prevalence of low serum sodium levels (<135 mmol/L) was high in both inpatients and outpatients (57% and 40%, respectively). The existence of serum sodium <135 mmol/L was associated with severe ascites, as indicated by high prevalence of refractory ascites, large fluid accumulation rate, frequent use of large-volume paracentesis, and impaired renal function, compared with normal serum sodium levels. Moreover, low serum sodium levels were also associated with greater frequency of hepatic encephalopathy, spontaneous bacterial peritonitis, and hepatorenal syndrome, but not gastrointestinal bleeding. Patients with serum sodium <130 mmol/L had the greatest frequency of these complications, but the frequency was also increased in patients with mild reduction in serum sodium levels (131-135 mmol/L). In conclusion, low serum sodium levels in cirrhosis are associated with severe ascites and high frequency of hepatic encephalopathy, spontaneous bacterial peritonitis, and hepatorenal syndrome. PMID- 17133459 TI - Genomics and complex liver disease: Challenges and opportunities. AB - The concept of genetic susceptibility in the contribution to human disease is not new. What is new is the emerging ability of the field of genomics to detect, assess, and interpret genetic variation in the study of susceptibility to development of disease. Deciphering the human genome sequence and the publication of the human haplotype map are key elements of this effort. However, we are only beginning to understand the contribution of genetic predisposition to complex liver disease through its interaction with environmental risk factors. In the coming decade, we anticipate the development of human studies to better dissect the genotype/phenotype relationship of complex liver diseases. This endeavor will require large, well-phenotyped patient populations of each disease of interest and proper study designs aimed at answering important questions of hepatic disease prognosis, pathogenesis, and treatment. Teamwork between patients, physicians, and genomics scientists can ensure that this opportunity leads to important biological discoveries and improved treatment of complex disease. PMID- 17133464 TI - JAK2 mutation: The best diagnostic tool for myeloproliferative disease in splanchnic vein thrombosis? PMID- 17133465 TI - Time to ban smoking in patients with chronic liver diseases. PMID- 17133466 TI - Fulminant hepatitis A: Disappearing, but not soon enough. PMID- 17133467 TI - Growth factors during HCV therapy may be "cost-effective", but are they "effective"? PMID- 17133468 TI - Smoking and increased severity of hepatic fibrosis in primary biliary cirrhosis: A cross validated retrospective assessment. AB - An epidemiological association between cigarette smoking and primary biliary cirrhosis (PBC) has been demonstrated. Our aim was to determine the relationship between smoking and severity of liver fibrosis at presentation in patients with PBC. All patients with PBC seen at the three major teaching hospitals of Case Western Reserve University between October 1998 and December 2005 were identified. Data obtained at the time of the first evaluation leading to the PBC diagnosis on 97 patients were collected. The cumulative number of cigarette packs smoked per year (pack-years) was calculated. Advanced histological disease was defined as Ludwig stages 3 or 4. Analyses were performed to determine associations between advanced histological disease, smoking and other variables related to liver fibrosis. Smoking history was more common (P = .0008) in patients with advanced histological disease at presentation compared to those with early disease. Among smokers, mean lifetime tobacco consumption was higher (P = .04) in cases with advanced histological disease at presentation (30 pack years) compared to cases with early disease (17 pack-years). Logistic regression demonstrated a significant association between a lifetime tobacco consumption of > or =10 pack-years and advanced histological disease at presentation (OR = 13.3). The association remained significant after adjusting for age, gender, and alcohol intake. The validity of these results was corroborated by cross validation in an independent confirmatory set of 172 patients with PBC. In conclusion, smoking may accelerate the progression of PBC. This could be induced by exposure to chemicals in cigarette smoke. PMID- 17133469 TI - Dynamic FDG-PET is useful for detection of cholangiocarcinoma in patients with PSC listed for liver transplantation. AB - Five to 15% of patients with primary sclerosing cholangitis (PSC) develop cholangiocarcinoma (CC) with a median survival of 5 to 7 months, an outcome not significantly improved by liver transplantation. However, if CC is found incidentally during the procedure or in the explanted liver, 5-year survival rates of 35% are reported. A noninvasive method to detect CC small enough to allow for intended curative surgery is needed. Unfortunately, computed tomography (CT) and ultrasonography (US) have poor sensitivity for detection of CC in PSC; however, positron emission tomography (PET) using 2-[18F]fluoro-2-deoxy-D-glucose (FDG) differentiates well between CC and nonmalignant tissue. We examined whether PET findings are valid using a blinded study design comparing pretransplantation FDG-PET results with histology of explanted livers. Dynamic FDG-PET was performed in 24 consecutive patients with PSC within 2 weeks after listing for liver transplantation and with no evidence of malignancy on CT, magnetic resonance imaging, or ultrasonography. The PET Center staff was blinded to clinical findings, and surgeons and pathologists were blinded to the PET results. Three patients had CC that was correctly identified by PET. PET was negative in 1 patient with high-grade hilar duct dysplasia. In 20 patients without malignancies, PET was false positive in 1 patient with epitheloid granulomas in the liver. In conclusion, dynamic FDG-PET appears superior to conventional imaging techniques for both detection and exclusion of CC in advanced PSC. FDG PET may be useful for screening for CC in the pretransplant evaluation of patients with PSC. PMID- 17133470 TI - Outcome of acute idiosyncratic drug-induced liver injury: Long-term follow-up in a hepatotoxicity registry. AB - A chronic adverse reaction may occur in some instances of drug-induced liver injury (DILI), even despite drug cessation. In our study, we obtained records from a Spanish registry and evaluated cases of DILI with biochemical evidence of long-term damage. Chronic outcome was defined as a persistent biochemical abnormality of hepatocellular pattern of damage more than 3 months after drug withdrawal or more than 6 months after cholestatic/mixed damage. Data on 28 patients with a chronic clinical evolution (mean follow-up 20 months) between November 1995 and October 2005 were retrieved (18 female; overall mean age 55 yr) and accounted for 5.7% of total idiosyncratic DILI cases (n = 493) submitted to the registry. The main drug classes were cardiovascular and central nervous system (28.5% and 25%, respectively), which, in contrast, represented only 9.8% and 13%, respectively, of all DILI cases. The most frequent causative drugs were amoxicillin-clavulanate (4 of 69 cases), bentazepam (3 of 7 cases), atorvastatin (2 of 7 cases), and captopril (2 of 5 cases). Patients with cholestatic/mixed injury (18 of 194 cases [9%]) were more prone to chronicity than patients with hepatocellular injury (10 of 240 cases; P < .031). In the case of chronic hepatocellular injury, 3 patients progressed to cirrhosis and 2 to chronic hepatitis. In the cholestatic/mixed group, liver biopsy indicated cirrhosis in 1 patient and ductal lesions in 3 patients. In conclusion, cholestatic/mixed type of damage is more prone to become chronic while, in the hepatocellular pattern, the severity is greater. Cardiovascular and central nervous system drugs are the main groups leading to chronic liver damage. PMID- 17133471 TI - IP-10 predicts viral response and therapeutic outcome in difficult-to-treat patients with HCV genotype 1 infection. AB - Plasma from 173 patients with HCV genotype 1 infection was analyzed for IP-10 levels prior to treatment with pegylated interferon-alpha-2a and ribavirin. Significantly lower IP-10 levels were observed in patients achieving a rapid viral response (RVR) (P < .0001), even in those with body mass index (BMI) > or = 25 kg/m2 (P = .004) and with baseline viral load > or = 2 million IU/mL (P = .001). Similarly, significantly lower IP-10 levels were observed in patients obtaining a sustained viral response (SVR) (P = .0002), including those having higher BMI (P < .05), higher viral load (P = .0005), and both higher BMI and viral load (P < .03). In multivariate logistic regression analyses, a low IP-10 value was independently predictive of both RVR and SVR. A baseline cutoff IP-10 value of 600 pg/mL yielded a negative predictive value (NPV) of 79% (19/24) for all genotype 1-infected patients, which was comparable with that observed using a reduction in HCV-RNA by at least 2 logs after 12 weeks of therapy (NPV 86%; 19/22); by combining the two, 30 of 38 patients (NPV 79%) potentially could have been spared unnecessary therapy. In patients having both higher BMI and viral load, cut-off levels of 150 and 600 pg/mL yielded a positive predictive value (PPV) of 71% and NPV of 100%, respectively. In conclusion, pretreatment IP-10 levels predict RVR and SVR in patients infected with HCV genotype 1, even in those with higher BMI and viral load. A substantial proportion of the latter patients may achieve SVR in spite of unfavorable baseline characteristics if their pretreatment IP-10 level is low. Thus, pretreatment IP-10 analysis may prove helpful in decision-making regarding pharmaceutical intervention. PMID- 17133472 TI - Serum amyloid A has antiviral activity against hepatitis C virus by inhibiting virus entry in a cell culture system. AB - Serum amyloid A (SAA) is an acute phase protein produced by the liver. SAA concentration increases markedly in the serum following inflammation and infection. Large increases in SAA concentration during the acute phase response suggest that SAA has a beneficial role in host defense. This study sought to determine the effect of SAA on hepatitis C virus (HCV) infectivity using retroviral particles pseudotyped with HCV envelope glycoproteins (HCVpp) and the recently developed cell culture system for HCV (HCVcc). SAA inhibited HCVpp and HCVcc infection in a dose-dependent manner by affecting an early step of the virus life cycle. Further characterization with HCVpp indicated that SAA blocks virus entry by interacting with the viral particle. In addition, the antiviral activity of SAA was strongly reduced when high-density lipoproteins (HDL) were coincubated with SAA. However, HDL had only a slight effect on the antiviral activity of SAA when HCVpp was first preincubated with SAA. Furthermore, analyses of SAA in sera of chronic HCV patients revealed the presence of variable levels of SAA with abnormally elevated concentrations in some cases. However, no obvious clinical correlation was found between SAA levels and HCV viral loads. In conclusion, our data demonstrate an antiviral activity for SAA and suggest a tight relationship between SAA and HDL in modulating HCV infectivity. PMID- 17133473 TI - Fibrosis in genotype 3 chronic hepatitis C and nonalcoholic fatty liver disease: Role of insulin resistance and hepatic steatosis. AB - Hepatic steatosis has been associated with fibrosis, but it is unknown whether the latter is independent of the etiology of fat infiltration. We analyzed the relationship between clinical characteristics, insulin resistance (HOMA-R) and histological parameters in 132 patients with "viral" steatosis caused by genotype 3 chronic hepatitis C (CHC-3) and 132 patients with "metabolic" steatosis caused by nonalcoholic fatty liver disease (NAFLD), matched by age, BMI, and degree of liver fat accumulation. Tests of liver function were comparable in the two study populations. The prevalence of features of insulin resistance was higher in NAFLD, as was HOMA-R (P = .008). Logistic regression analysis confirmed that steatosis was associated with a high viral load and low serum cholesterol in CHC 3, and with high aminotransferase, glucose, ferritin and hypertriglyceridemia in NAFLD. At univariate analysis, advanced fibrosis was associated with steatosis in NAFLD, but not in CHC-3. Other parameters related to fibrosis severity were HOMA R and a low platelet count in CHC-3, and high aminotransferases, HOMA-R, ferritin and low HDL-cholesterol in NAFLD. On multivariate analysis, only low platelet count (OR = 0.78; 95% CI, 0.67-0.92) and HOMA-R (OR = 2.98; 1.13-7.89) were independent predictors of advanced fibrosis in CHC-3. In NAFLD, severe fibrosis was predicted by fat grading (OR = 3.03; 1.41-6.53), ferritin (OR = 1.13; 1.03 1.25) and HOMA-R (OR = 1.16; 1.02-1.31). In conclusion, insulin resistance is an independent predictor of advanced fibrosis in both NAFLD and CHC-3, but the extent of steatosis contributes to advanced disease only in NAFLD. Virus-induced hepatic steatosis as seen in CHC-3 does not contribute significantly to liver fibrosis. PMID- 17133474 TI - Characterization of HBV DNA+/HBsAg- blood donors in Poland identified by triplex NAT. AB - Nucleic acid testing (NAT) for hepatitis B virus (HBV) has been performed in Poland since 2005 on samples seronegative for hepatitis B surface antigen (HBsAg), anti-hepatitis C virus (anti-HCV), and anti-human immunodeficiency virus (anti-HIV). Tools included 24-donation pool testing (PT) using Cobas Amplicor or in individual donations (ID) by Procleix Ultrio. Seven of 761,666 (1:108,800) and 21/250,191 (1:11,900) HBV DNA-positive donations were identified and confirmed by alternative methods. HBV DNA load ranged between 11.6 and 4.6 x 10(4) IU/mL in 11 samples and could not be quantified in 17 samples. HBV genotypes A (56%) and D (4%) were found. The analysis of combined results from index, follow-up, and look back samples identified four groups: (1) Two cases tested HBsAg positive with alternative, more sensitive, assays; (2) Four cases were in the pre seroconversion window period; (3) Eight cases had a fluctuating pattern of HBV DNA and anti-HBs detection (recovered infection); and (4) twelve cases carried anti-HBc without anti-HBs, which might correspond to either chronic or recovered "occult" HBV infection. One donor with no HBV markers in the follow-up was excluded, and another was in the window period preceding anti-HBs. HBV NAT identified more confirmed positive donors than HCV or HIV NAT, and 1:250,000 could not be detected by anti-HBc screening. Serological and molecular studies on follow-up and look-back samples are important to classify donors. In conclusion, further studies are needed to determine whether the considerably higher yield of HBV DNA detection obtained with individual donation screening improves blood safety compared with anti-HBc screening. PMID- 17133475 TI - Entecavir resistance is rare in nucleoside naive patients with hepatitis B. AB - Comprehensive monitoring of genotypic and phenotypic antiviral resistance was performed on 673 entecavir (ETV)-treated nucleoside naive hepatitis B virus (HBV) patients. ETV reduced HBV DNA levels to undetectable by PCR (<300 copies/mL, <57 IU/mL) in 91% of hepatitis B e antigen (HBeAg)-positive and -negative patients by Week 96. Thirteen percent (n = 88) of the comparator lamivudine (LVD)-treated patients experienced a virologic rebound (> or =1 log increase from nadir by PCR) in the first year, with 74% of these having LVD resistance (LVDr) substitutions evident. In contrast, only 3% (n = 22) of ETV-treated patients exhibited virologic rebound by Week 96. Three ETV rebounds were attributable to LVDr virus present at baseline, with one having a S202G ETV resistance (ETVr) substitution emerge at Week 48. None of the other rebounding patients had emerging genotypic resistance or loss of ETV susceptibility. Genotyping all additional ETV patients with PCR-detectable HBV DNA at Weeks 48, 96, or end of dosing identified seven additional patients with LVDr substitutions, including one with simultaneous emergence of LVDr/ETVr. Generally, ETV patients with LVDr were detectable at baseline (8/10) and most subsequently achieved undetectable HBV DNA levels on ETV therapy (7/10). No other emerging substitutions identified decreased ETV susceptibility. In conclusion, ETVr emergence in ETV-treated nucleoside naive patients over a 2-year period is rare, occurring in two patients with LVDr variants. These findings suggest that the rapid, sustained suppression of HBV replication, combined with a requirement for multiple substitutions, creates a high genetic barrier to ETVr in nucleoside naive patients. PMID- 17133476 TI - H. pylori and platelet counts. PMID- 17133477 TI - Surprises from the crystal structure of the hepatitis C virus NS2-3 protease. PMID- 17133478 TI - Hepatitis B virus resistance to entecavir in nucleoside naive patients: Does it exist? PMID- 17133479 TI - Diabetes and hepatocellular carcinoma. PMID- 17133480 TI - Monocrotaline promotes transplanted cell engraftment and advances liver repopulation in rats via liver conditioning. AB - Disruption of the hepatic endothelial barrier or Kupffer cell function facilitates transplanted cell engraftment in the liver. To determine whether these mechanisms could be activated simultaneously, we studied the effects of monocrotaline, a pyrollizidine alkaloid, with reported toxicity in liver sinusoidal endothelial cells and Kupffer cells. The effects of monocrotaline in Fischer 344 rats were examined by tissue morphology, serum hyaluronic acid levels, and liver tests (endothelial and hepatocyte injury) or incorporation of carbon and (99m)Tc-sulfur colloid (Kupffer cell damage). To study changes in cell engraftment and liver repopulation, Fischer 344 rat hepatocytes were transplanted into syngeneic dipeptidyl peptidase IV-deficient rats followed by histological assays. We observed extensive endothelial injury without Kupffer cell or hepatocyte damage in monocrotaline-treated rats. Monocrotaline enhanced transplanted cell engraftment without changes in transplanted cell numbers or induction of proliferation in native hepatocytes over 3 months. In monocrotaline treated rats, transplanted cells integrated into the liver parenchyma and survived in vascular spaces. To determine whether native hepatocytes suffered inapparent damage after monocrotaline, we introduced further liver injury with carbon tetrachloride subsequent to cell transplantation. Monocrotaline sensitized the liver to carbon tetrachloride-induced necrosis, which advanced transplanted cell proliferation, leading to significant liver repopulation. During this process, we observed proliferation of bile duct cells and small epithelial cells, although transplanted hepatocytes did not appear to reconstitute bile ducts. The studies showed that perturbation of multiple liver cell compartments by monocrotaline promoted transplanted cell engraftment and proliferation. In conclusion, development of drugs with monocrotaline-like effects will help advance liver cell therapy. PMID- 17133481 TI - Role of neutrophils in a mouse model of halothane-induced liver injury. AB - Drug-induced liver injury (DILI) is a major safety concern in drug development. Its prediction and prevention have been hindered by limited knowledge of the underlying mechanisms, in part the result of a lack of animal models. We developed a mouse model of halothane-induced liver injury and characterized the mechanisms accounting for tissue damage. Female and male Balb/c, DBA/1, and C57BL/6J mice were injected intraperitoneally with halothane. Serum levels of alanine aminotransferase and histology were evaluated to determine liver injury. Balb/c mice were found to be the most susceptible strain, followed by DBA/1, with no significant hepatotoxicity observed in C57BL/6J mice. Female Balb/c and DBA/1 mice developed more severe liver damage compared with their male counterparts. Bioactivation of halothane occurred similarly in all three strains based on detection of liver proteins adducted by the reactive metabolite. Mechanistic investigations revealed that hepatic message levels of tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta); IL-6, and IL-8 were significantly higher in halothane-treated Balb/c mice compared to DBA/1 and C57BL/6J mice. Moreover, a higher number of neutrophils were recruited into the liver of Balb/c mice upon halothane treatment compared with DBA/1, with no obvious neutrophil infiltration detected in C57BL/6J mice. Neutrophil depletion experiments demonstrated a crucial role for these cells in the development of halothane-induced liver injury. The halothane-initiated hepatotoxicity and innate immune response-mediated escalation of tissue damage are consistent with events that occur in many cases of DILI. In conclusion, our model provides a platform for elucidating strain-based and gender-based susceptibility factors in DILI development. PMID- 17133482 TI - JunD is a profibrogenic transcription factor regulated by Jun N-terminal kinase independent phosphorylation. AB - JunD is implicated in the regulation of hepatic stellate cell (HSC) activation and liver fibrosis via its transcriptional regulation of the tissue inhibitor of metalloproteinases-1 (TIMP-1) gene. In the present study we found in vivo evidence of a role for JunD in fibrogenesis. Expression of JunD was demonstrated in alpha-SMA-positive activated HSCs of fibrotic rodents and human livers. The junD-/- mice were protected from carbon tetrachloride-induced fibrosis. The livers of injured junD-/- mice displayed significantly reduced formation of fibrotic crosslinked collagen and a smaller number of alpha-SMA-positive HSCs compared with those of wild-type (wt) mice. Hepatic TIMP-1 mRNA expression in injured junD-/- mice was 78% lower and in culture activated junD-/- HSCs was 50% 80% lower than that in wt mice. In examining the signal transduction mechanisms that regulate JunD-dependent TIMP-1 expression, we found a role for phosphorylation of the Ser100 residue of JunD but ruled out JNK as a mediator of this event, suggesting ERK1/2 is utilized. In conclusion, a signaling pathway for the development of fibrosis involves the regulation of TIMP-1 expression by phosphorylated JunD. PMID- 17133483 TI - STAT1 inhibits liver fibrosis in mice by inhibiting stellate cell proliferation and stimulating NK cell cytotoxicity. AB - Liver fibrosis, a common scarring response to chronic liver injury, is a precursor to cirrhosis and liver cancer. Here, we identified signal transducer and activator of transcription 1 (STAT1) as an important negative regulator in liver fibrosis. Our findings show that disruption of the STAT1 gene accelerated liver fibrosis and hepatic stellate cell (HSC) proliferation in an in vivo model of carbon tetrachloride (CCl4)-induced liver fibrosis. In vitro treatment with IFN-gamma inhibited proliferation and activation of wild-type HSCs, but not STAT1 /- HSCs. Moreover, compared to wild-type cells, cellular proliferation stimulated by serum or platelet-derived growth factor (PDGF) was enhanced and accelerated in STAT1-/- HSCs, which was partially mediated via elevated PDGF receptor beta expression on such cells. Polyinosinic-polycytidylic acid (poly I:C) or IFN-gamma treatment inhibited liver fibrosis in wild-type mice but not in STAT1-/- mice. Induction of NK cell killing of activated HSCs by poly I:C was attenuated in STAT1-/- mice compared to wild-type mice, which was likely due to reduced NKG2D and TRAIL expression on STAT1-/- NK cells. Finally, activation of TGF-beta/Smad3 signaling pathway was accelerated, whereas induction of Smad7 was diminished in the liver of STAT1-/- mice after CCl4 administration compared to wild-type mice. In conclusion, activation of STAT1 attenuates liver fibrosis through inhibition of HSC proliferation, attenuation of TGF-beta signaling, and stimulation of NK cell killing of activated HSCs. STAT1 could be a new therapeutic target for treating liver fibrosis. PMID- 17133484 TI - Engineered measles virus as a novel oncolytic viral therapy system for hepatocellular carcinoma. AB - The oncolytic measles virus Edmonston strain (MV-Edm), a nonpathogenic virus targeting cells expressing abundant CD46, selectively destroys neoplastic tissue. Clinical development of MV-Edm would benefit from noninvasive monitoring strategies to determine the speed and extent of the spread of the virus in treated patients and the location of virus-infected cells. We evaluated recombinant MV-Edm expressing carcinoembryonic antigen (CEA) or the human sodium iodide symporter (hNIS) for oncolytic potential in hepatocellular carcinoma (HCC) and efficiency in tracking viruses in vivo by noninvasive monitoring. CD46 expression in human HCC and primary hepatocytes was assessed by flow cytometry and immunohistochemistry. Infectivity, syncytium formation, and cytotoxicity of recombinant MV-Edm in HCC cell lines were evaluated by fluorescence microscopy, crystal violet staining, and the MTS assay. Transgene expression in HCC cell lines after infection with recombinant MV-Edm in vitro and in vivo was assessed by CEA concentration, 125I-uptake, and 123I-imaging studies. Toxicology studies were performed in Ifnar(KO)xCD46 transgenic mice. The CD46 receptor was highly expressed in HCC compared to nonmalignant hepatic tissue. Recombinant MV-Edm efficiently infected HCC cell lines, resulting in extensive syncytium formation followed by cell death. Transduction of HCC cell lines and subcutaneous HCC xenografts with recombinant MV-Edm resulted in high-level expression of transgenes in vitro and in vivo. MV-Edm was nontoxic in susceptible mice. Intratumoral and intravenous therapy with recombinant MV-Edm resulted in inhibition of tumor growth and prolongation of survival with complete tumor regression in up to one third of animals. In conclusion, engineered MV-Edm may be a potent and novel cancer gene therapy system for HCC. MV-Edm expressing CEA or hNIS elicited oncolytic effects in human HCC cell lines in vitro and in vivo, enabling the spread of the virus to be monitored in a noninvasive manner. PMID- 17133485 TI - Reg2 inactivation increases sensitivity to Fas hepatotoxicity and delays liver regeneration post-hepatectomy in mice. AB - Reg2/RegIIIbeta is the murine homologue of the human secreted HIP/PAP C-type lectin. HIP/PAP transgenic mice were protected against acetaminophen-induced acute liver failure and were stimulated to regenerate post-hepatectomy. To assess the role of Reg2, we used Reg2-/- mice in a model of fulminant hepatitis induced by Fas and in the post-hepatectomy regeneration. Within 4 hours of J0-2 treatment (0.5 microg/g), only 50% of the Reg2-/- mice were alive but with an increased sensitivity to Fas-induced oxidative stress and a decreased level of Bcl-xL. In contrast, HIP/PAP transgenic mice were resistant to Fas, with HIP/PAP serving as a sulfhydryl buffer to slow down decreases in glutathione and Bcl-xL. In Reg2-/- mice, liver regeneration was markedly impaired, with 29% mortality and delay of the S-phase and the activation of ERK1/2 and AKT. Activation of STAT3 began on time at 3 hours but persisted strongly up to 72 hours despite significant accumulation of SOCS3. Thus, Reg2 deficiency induced exaggerated IL-6/STAT-3 activation and mito-inhibition. Because the Reg2 gene was activated between 6 and 24 hours after hepatectomy in wild-type mice, Reg2 could mediate the TNF-alpha/IL 6 priming signaling by exerting a negative feed-back on STAT3/IL-6 activation to allow the hepatocytes to progress through the cell cycle. In conclusion, Reg2 deficiency enhanced liver sensitivity to Fas-induced oxidative stress and delayed liver regeneration with persistent TNF-alpha/IL6/STAT3 signaling. In contrast, overexpression of human HIP/PAP promoted liver resistance to Fas and accelerated liver regeneration with early activation/deactivation of STAT3. Reg2/HIP/PAP is therefore a critical mitogenic and antiapoptotic factor for the liver. PMID- 17133486 TI - Hepatic precursors derived from murine embryonic stem cells contribute to regeneration of injured liver. AB - We established an efficient system for differentiation, expansion and isolation of hepatic progenitor cells from mouse embryonic stem (ES) cells and evaluated their capacity to repopulate injured liver. Using mouse ES cells transfected with the green fluorescent protein (GFP) reporter gene regulated by albumin (ALB) enhancer/promoter, we found that a serum-free chemically defined medium supports formation of embryoid bodies (EBs) and differentiation of hepatic lineage cells in the absence of exogenous growth factors or feeder cell layers. The first GFP+ cells expressing ALB were detected in close proximity to "beating" myocytes after 7 days of EB cultures. GFP+ cells increased in number, acquired hepatocyte-like morphology and hepatocyte-specific markers (i.e., ALB, AAT, TO, and G6P), and by 28 days represented more than 30% of cells isolated from EB outgrowths. The FACS purified GFP+ cells developed into functional hepatocytes without evidence of cell fusion and participated in the repairing of diseased liver when transplanted into MUP-uPA/SCID mice. The ES cell-derived hepatocytes were responsive to normal growth regulation and proliferated at the same rate as the host hepatocytes after an additional growth stimulus from CCl(4)-induced liver injury. The transplanted GFP+ cells also differentiated into biliary epithelial cells. In conclusion, a highly enriched population of committed hepatocyte precursors can be generated from ES cells in vitro for effective cell replacement therapy. PMID- 17133487 TI - Oxidative-stress and IL-6 mediate the fibrogenic effects of [corrected] Kupffer cells on stellate cells. AB - The impact of Kupffer cells (KCs) on the hepatic stellate cell (HSC) fibrogenic response was examined in an in vitro coculture model of primary KCs and HSCs. Coculture with KCs induced a more activated phenotype and greater proliferation compared to HSC cultured alone. Similar results were obtained on Matrigel which maintains HSCs quiescent. The effect of KCs on HSC collagen I involved transcriptional regulation, as determined by nuclear in vitro transcription run on assays, promoter studies, and Northern blot analysis, while stability of the COL1A1 and COL1A2 mRNA were similar. The minimal COL1A1 and COL1A2 promoter regions responsible for the KC effects were localized to the -515 and -378 base pair (bp) regions, respectively. Intracellular and extracellular collagen I protein, H2O2, and IL-6 increased in a time-dependent fashion, especially for HSCs in coculture. Catalase prevented these effects as well as the transactivation of both collagen promoters. The rate of collagen I protein synthesis and intracellular collagen I degradation remained similar but the t(1/2) of the secreted collagen I was lower for HSC in coculture. MMP13, a protease that degrades extracellular collagen I, decreased in the cocultures, while TIMP1, a MMP13 inhibitor, increased; and these effects were prevented by catalase, anti-IL-6, and siRNA-IL-6. Cocultured HSC showed elevated phosphorylation of p38 which when inhibited by catalase, anti-IL-6, and siRNA-IL 6 it blocked TIMP1 upregulation and collagen I accumulation. In conclusion, these results unveil a novel dual mechanism mediated by H2O2 and IL-6 by which KCs may modulate the fibrogenic response in HSCs. PMID- 17133488 TI - Portopulmonary hypertension: Results from a 10-year screening algorithm. AB - Portopulmonary hypertension (POPH) is the elevation of pulmonary artery pressure due to increased resistance to pulmonary blood flow in the setting of portal hypertension. Increased mortality has occurred with attempted liver transplantation in such patients and thus, screening for POPH is advised. We examined the relationship between screening echocardiography and right heart catheterization determinations of pressure, flow, volume, and resistance. A prospective, echocardiography-catheterization algorithm was followed from 1996 to 2005. Consecutive transplantation candidates underwent Doppler echocardiography to determine right ventricular systolic pressure (RVSP). Of 1,235 patients, 101 with RVSP >50 mm Hg underwent catheterization to measure mean pulmonary artery pressure (MPAP), flow via cardiac output (CO), central volume via pulmonary artery occlusion pressure (PAOP), and resistance via calculated pulmonary vascular resistance (PVR). Bland-Altman analysis suggested marked discordance between echocardiography-derived RVSP and catheterization results. All-cause pulmonary hypertension (MPAP >25 mm Hg) was documented in 90/101 (90%) patients. Using current pressure and resistance diagnostic guidelines (MPAP >25 mm Hg, PVR > or =240 dynes/s/cm(-5)), POPH was documented in 66/101 (65%) patients. Elevated MPAP was due to increased CO and/or PAOP in 35/101 (35%) patients with normal resistance (PVR <240 dynes/s/cm(-5)). The transpulmonary gradient (MPAP-PAOP) further characterized POPH in the presence of increased volume. Model for end stage liver disease (MELD) scores correlated poorly with MPAP and PVR. In conclusion, right heart catheterization is necessary to confirm POPH and frequently identifies other reasons for pulmonary hypertension (e.g., high flow and increased central volume) in liver transplantation candidates. Severity of POPH correlates poorly with MELD scores. PMID- 17133489 TI - Fulminant hepatitis A virus infection in the United States: Incidence, prognosis, and outcomes. AB - Acute liver failure (ALF) due to hepatitis A virus (HAV) infection is an uncommon but potentially lethal illness. The aim of this study was to identify readily available laboratory and clinical features associated with a poor prognosis among ALF patients with HAV infection. The presenting features of 29 adults with anti HAV IgM positive ALF enrolled in the ALFSG_between 1998 and 2005 were reviewed. The HAV patients listed for transplantation by UNOS were also reviewed. Acute HAV accounted for 3.1% of patients enrolled in the ALFSG. At 3 weeks follow-up, 16 had spontaneously recovered (55%), 9 underwent transplantation (31%), and 4 had died (14%). A prognostic model incorporating 4 presenting features (serum ALT <2,600 IU/L, creatinine >2.0 mg/dL, intubation, pressors) had an AUROC for transplant/death of 0.899 which was significantly better than the King's College criteria (0.623, P = .018) and MELD scores (0.707, P = .0503). Between 1988 and 2005, the frequency of patients requiring liver transplantation for HAV in the UNOS database significantly decreased from 0.7 % to 0.1% (P < .001). In addition, the proportion of HAV cases enrolled in the ALFSG significantly decreased from 5% to 0.8% (P = .007). In conclusion, the frequency of HAV patients enrolling in the ALFSG and being listed for liver transplantation in the United States has declined in parallel. A prognostic index consisting of 4 clinical and laboratory features predicted the likelihood of transplant/death significantly better than other published models suggesting that disease specific prognostic models may be of value in non-acetaminophen ALF. PMID- 17133490 TI - Cost-effectiveness of hematologic growth factors for anemia occurring during hepatitis C combination therapy. AB - In hepatitis C virus (HCV)-infected patients who develop anemia during combination therapy, erythropoietic growth factors maintain higher drug treatment levels compared to ribavirin dose reduction, which may lead to an increase in treatment response rates. This study estimated the cost-effectiveness of growth factor therapy in maintaining anemic HCV-infected patients on target drug levels during combination therapy. A decision analysis using a Markov model was developed with 7 health states: Sustained viral response, chronic HCV, compensated cirrhosis, decompensated cirrhosis, hepatocellular carcinoma, liver transplantation, and death. Data sources included population-based studies of growth factor therapy, previously published estimates of costs and natural history of hepatitis C, and recent prospective studies. Our reference case was a 45-year-old Caucasian man with HCV infection (genotype 1, 2, or 3) who developed anemia while undergoing combination therapy with ribavirin and pegylated interferon. We compared growth factor injections (darbepoetin alpha or epoetin alpha) during combination therapy with standard ribavirin dose reduction. Compared to a ribavirin dose reduction strategy, the cost of darbepoetin per additional quality-adjusted life-year was 34,793 dollars for genotype 1 and 33,832 dollars for genotypes 2 or 3 versus 60,600 dollars and 64,311 dollars for epoetin. For all genotypes, the results were sensitive to changes in the cure rates of HCV therapy, the utility of chronic HCV, the costs of growth factors, and the age at which therapy is begun. In conclusion, use of erythropoietic growth factors, specifically darbepoetin, for patients with anemia occurring during HCV combination therapy appears to be cost-effective for genotypes 1, 2, or 3. PMID- 17133491 TI - Intrahepatic virus-specific IL-10-producing CD8 T cells prevent liver damage during chronic hepatitis C virus infection. AB - CD8 T cell killing of hepatitis C virus (HCV)-infected hepatocytes is thought to contribute to liver damage during chronic HCV infection, whereas the participation of HCV-nonspecific immune cells is unclear. To visualize the spatial relationship of HCV-specific CD8 T cells with parenchymal target cells, and to examine their local functional activity in relation to hepatocellular necrosis and fibrosis, we used HLA tetramers and confocal microscopy in biopsies from 23 HLA-A2 or HLA-B7 patients with chronic HCV infection. Intrahepatic tetramer+ (HCV-specific) CD8 T cells protected from hepatic necroinflammatory disease activity, independently of age, gender, viral load, and viral genotype. Indeed, tetramer+ cells were scattered in the liver within regions of weak fibrosis (low laminin expression) and low hepatocellular apoptosis (TUNEL method), and expressed IL-10 but not IFNgamma. By contrast, tetramer-negative CD8 T cells were associated with active necroinflammatory liver disease, colocalized with strong laminin expression and hepatocellular apoptosis, and expressed more frequently IFNgamma than IL-10. Overall, liver regions harboring HCV-specific CD8 T cells tended to be healthier than areas containing only inflammatory cells of undefined specificity. In conclusion, HCV-specific IL-10-producing CD8 T cells, although not cytotoxic and unable to control viral replication, can attenuate hepatocellular necrosis, liver fibrosis, and inflammation mediated by bystander T cells, and may thus represent antigen-induced regulatory CD8 T cells. Therapeutic modulation of the intrahepatic balance between specific and bystander CD8 T cells might be beneficial in patients with chronic hepatitis C. PMID- 17133492 TI - Prevention of hepatocellular carcinoma recurrence with alpha-interferon after liver resection in HCV cirrhosis. AB - Tumor recurrence after resection of hepatocellular carcinoma (HCC) can occur early (<2 years) or late (>2 years) as metastases or de novo tumors. Interferon (IFN) has the potential for chemoprevention against hepatitis C virus (HCV) related cirrhosis. A predetermined group of 150 HCV RNA-positive patients undergoing resection of early- to intermediate-stage HCC was stratified into 80 HCV-pure (hepatitis B anticore antibody [anti-HBc]-negative) and 70 mixed HCV+hepatitis B virus (HBV) (anti-HBc-positive) groups, then randomized to IFN alpha (3 million units 3 times every week for 48 weeks [n = 76]) versus control (n = 74). The primary end point was recurrence-free survival (RFS); secondary end points were disease-specific and overall survival. Intention-to-treat and subgroup analysis on adherent patients were conducted. Treatment effects on early/late recurrences were assessed using multiple Cox regression analysis. No patient experienced life-threatening adverse events. There were 28 adherent patients (37%). After 45 months of median follow-up, overall survival was 58.5%, and no significant difference in RFS was detectable between the two study arms (24.3% vs. 5.8%; P = .49). HCC recurred in 100 patients (48 IFN-treated, 52 controls), with a 50% reduction in late recurrence rate in the treatment arm. HCC multiplicity and vascular invasion were significantly related to recurrence (P = .01 and .0003). After viral status stratification, while no treatment effect was apparent in the mixed HCV+HBV population and on early recurrences (72 events), there was a significant benefit on late recurrences (28 events) in HCV-pure patients adherent to treatment (HR: 0.3; 95% CI: 0.09-0.9; P = .04). In conclusion, IFN does not affect overall prevention of HCC recurrence after resection, but it may reduce late recurrence in HCV-pure patients receiving effective treatment. PMID- 17133493 TI - Aquaporin-1 and aquaporin-2 urinary excretion in cirrhosis: Relationship with ascites and hepatorenal syndrome. AB - Several experimental models of cirrhosis have shown dysregulation of renal aquaporins in different phases of liver disease. We investigated the urinary excretion of both aquaporin-1 and aquaporin-2 in patients with cirrhosis at different stages of the disease. Twenty-four-hour urine was collected from 11 healthy volunteers, 13 patients with compensated cirrhosis (without ascites), and 20 patients with decompensated cirrhosis (11 with ascites without renal failure and 9 with hepatorenal syndrome). Aquaporin-1 and aquaporin-2 excretion was analyzed by immunoblotting. Urinary aquaporin-2 excretion was reduced in patients with cirrhosis compared to healthy subjects. A progressive decrease in urinary aquaporin-2 excretion was observed as the severity of cirrhosis increased, from compensated cirrhosis to cirrhosis with ascites and hepatorenal syndrome. Patients with hyponatremia had lower urinary aquaporin-2 excretion than patients without hyponatremia. Vasopressin plasma level did not correlate with aquaporin-2 excretion. There were no differences between healthy subjects and patients with cirrhosis with or without ascites in urinary excretion of aquaporin-1, but urinary aquaporin-1 excretion of those with hepatorenal syndrome was extremely low. In conclusion, patients with cirrhosis appear to exhibit a decreased abundance of renal aquaporin-2 and therefore lower water permeability in the collecting tubules. This may represent an adaptive renal response to sodium retention, with expansion of extracellular fluid volume and dilutional hyponatremia observed in those who have cirrhosis with ascites. Finally, aquaporin-1 does not appear to play a role in the progressive dysregulation of extracellular fluid volume in cirrhosis. PMID- 17133494 TI - Laparoscopic liver surgery: Shifting the management of liver tumors. AB - Laparoscopic liver surgery has evolved rapidly over the past 5 years in a select number of centers. The growing experience with these procedures has resulted in a shift in the diagnostic and therapeutic approach to common liver tumors. The fact that resection of benign and malignant hepatic masses can now be accomplished laparoscopically with relatively low morbidity has influenced the decision-making process for physicians involved in the diagnosis and management of these lesions. For example, should a gastroenterologist or hepatologist seeing a 32-year-old woman with an asymptomatic 4 cm hepatic lesion that is radiologically indeterminate for adenoma or focal nodular hyperplasia (FNH): (1) continue to observe with annual computed tomography/magnetic resonance imaging (CT/MRI) scans, (2) subject the patient to a liver biopsy, or (3) refer for laparoscopic resection? For a solitary malignant liver tumor in the left lateral segment, should laparoscopic resection be considered the new standard of care, assuming the surgeon can perform the operation safely? We present current data and representative case studies on the use of laparoscopic liver resection at 2 major medical centers in the United States. We propose that surgical engagement defined by the managing physician's decision to proceed with a surgical intervention is increasingly affected by the availability of, and experience with, laparoscopic liver resection. PMID- 17133495 TI - Pituitary tumor transforming gene 1 in the liver. PMID- 17133497 TI - A picture says more than a thousand words: Structural insights into hepatitis C virus translation initiation. PMID- 17133498 TI - The protein kinase IKKepsilon can inhibit HCV expression independently of IFN and its own expression is downregulated in HCV-infected livers. AB - During a viral infection, binding of viral double-stranded RNAs (dsRNAs) to the cytosolic RNA helicase RIG-1 leads to recruitment of the mitochondria-associated Cardif protein, involved in activation of the IRF3-phosphorylating IKKepsilon/TBK1 kinases, interferon (IFN) induction, and development of the innate immune response. The hepatitis C virus (HCV) NS3/4A protease cleaves Cardif and abrogates both IKKepsilon/TBK1 activation and IFN induction. By using an HCV replicon model, we previously showed that ectopic overexpression of IKKepsilon can inhibit HCV expression. Here, analysis of the IKKepsilon transcriptome profile in these HCV replicon cells showed induction of several genes associated with the antiviral action of IFN. Interestingly, IKKepsilon still inhibits HCV expression in the presence of neutralizing antibodies to IFN receptors or in the presence of a dominant negative STAT1alpha mutant. This suggests that good IKKepsilon expression levels are important for rapid activation of the cellular antiviral response in HCV-infected cells, in addition to provoking IFN induction. To determine the physiological importance of IKKepsilon in HCV infection, we then analyzed its expression levels in liver biopsy specimens from HCV-infected patients. This analysis also included genes of the IFN induction pathway (RIG-I, MDA5, LGP2, Cardif, TBK1), and three IKKepsilon induced genes (IFN-beta, CCL3, and ISG15). The results show significant inhibition of expression of IKKepsilon and of the RNA helicases RIG-I/MDA5/LGP2 in the HCV-infected patients, whereas expression of TBK1 and Cardif was not significantly altered. In conclusion, given the antiviral potential of IKKepsilon and of the RNA helicases, these in vivo data strongly support an important role for these genes in the control of HCV infection. PMID- 17133499 TI - Impact of disease severity on outcome of antiviral therapy for chronic hepatitis C: Lessons from the HALT-C trial. AB - In patients with chronic hepatitis C, advanced fibrosis and cirrhosis are associated with lower rates of sustained virologic response (SVR) to interferon (IFN)-based therapy. In this study, we assessed virologic response to retreatment with peginterferon alfa-2a and ribavirin (RBV), as a function of the baseline fibrosis score (Ishak staging) and platelet count, in 1,046 patients enrolled in the Hepatitis C Antiviral Long-term Treatment against Cirrhosis (HALT-C) Trial. All patients had failed prior treatment with IFN or peginterferon +/- RBV and had Ishak fibrosis scores > or = 3. Four groups of patients with increasingly severe liver disease were compared: (A) bridging fibrosis (Ishak 3 and 4) with platelet counts >125,000/mm3 (n = 559); (B) bridging fibrosis with platelet counts < or =125,000/mm3 (n = 96); (C) cirrhosis (Ishak 5 and 6) with platelet counts >125,000/mm3 (n = 198); and (D) cirrhosis with platelet counts < or =125,000/mm3 (n = 193). SVR rates were 23%, 17%, 10%, and 9% in groups A, B, C, and D, respectively (P < .0001 for trend). Reduction in SVR as a function of increasingly severe disease was independent of age, percent African American, HCV genotype, HCV level, and type of prior therapy. Dose reduction lowered SVR frequencies, but to a lesser extent than disease severity. By logistic regression, cirrhosis (P < .0001) was the major determinant that impaired virologic response, independent of dose reduction or platelet count. In conclusion, disease severity is a major independent determinant of rate of SVR in patients with advanced chronic hepatitis C. New strategies are needed to optimize antiviral therapy in these "difficult-to-cure" patients. PMID- 17133500 TI - Assessing the role of DRD5 and DYT1 in two different case-control series with primary blepharospasm. AB - Primary blepharospasm is a common adult-onset focal dystonia. Polymorphisms of the genes encoding TorsinA (DYT1) and the D5 dopamine receptor (DRD5) have previously been associated with lifetime risk for focal dystonia. We describe here experiments testing common variability within these two genes in two independent cohorts of Italian and North American patients with primary blepharospasm. We have failed to identify a consistent association with disease in the two patient groups examined here; however, analysis of the Italian group reveals an association with the same risk genotype in DYT1 as previously described in an Icelandic population. We have also found global significant DYT1 haplotype differences between patients and controls in the Italian series. These data suggest that further examination is warranted of the role genetic variability at this locus plays in the risk for primary dystonia. PMID- 17133501 TI - Sfrp5 is not essential for axis formation in the mouse. AB - Secreted frizzled related protein (Sfrp) genes encode extracellular factors that can modulate Wnt signaling. During early post-implantation mouse development Sfrp5 is expressed in the anterior visceral endoderm (AVE) and the ventral foregut endoderm. The AVE is important in anterior-posterior axis formation and the ventral foregut endoderm contributes to multiple gut tissues. Here to determine the essential role of Sfrp5 in early mouse development we generated Sfrp5-deficient mice by gene targeting. We report that Sfrp5-deficient mice are viable and fertile. To determine whether the absence of an axis phenotype might be due to genetic redundancy with Dkk1 in the AVE we generated Sfrp5;Dkk1 double mutant mice. AVE development and primitive streak formation appeared normal in Sfrp5(-/-);Dkk1(-/-) embryos. These results indicate that Sfrp5 is not essential for axis formation or foregut morphogenesis in the mouse and also imply that Sfrp5 and Dkk1 together are not essential for AVE development. PMID- 17133502 TI - Progression of tremor and ataxia in male carriers of the FMR1 premutation. AB - Premutation alleles of the fragile X mental retardation 1 (FMR1) gene give rise to a late-onset movement disorder, fragile X-associated tremor/ataxia syndrome (FXTAS), characterized by progressive intention tremor and gait ataxia, with associated dementia and global brain atrophy. The natural history of FXTAS is largely unknown. To address this issue, a family-based, retrospective, longitudinal study was conducted with a cohort of 55 male premutation carriers. Analysis of the progression of the major motor signs of FXTAS, tremor and ataxia, shows that tremor usually occurs first, with median onset at approximately 60 years of age. From the onset of the initial motor sign, median delay of onset of ataxia was 2 years; onset of falls, 6 years; dependence on a walking aid, 15 years; and death, 21 years. Preliminary data on life expectancy are variable, with a range from 5 to 25 years. PMID- 17133503 TI - Accuracy of liver stiffness measurement for the diagnosis of cirrhosis in patients with chronic liver diseases. AB - A proper diagnosis of cirrhosis is essential for the management of patients with chronic liver diseases. We assessed the accuracy of liver stiffness measurement by Fibroscan for the diagnosis of cirrhosis in 1,257 patients with chronic liver diseases of various causes enrolled in a prospective multicenter study as well as clarified causes of discrepancies between liver histology and Fibroscan. One hundred thirty-two patients had unsuitable biopsy specimens, and 118 had unreliable liver stiffness measurements. Because 232 patients overlapped with a previous study, analysis was performed in the 775 new patients then derived in the whole population (1,007; 165 cirrhosis). Diagnostic accuracy was assessed by receiver operator curve (ROC) analysis. Liver samples were re-analyzed in case of discrepancies. The area under the ROC (AUROC) was 0.95 (95% CI, 0.93-0.96) for the diagnosis of cirrhosis in either 775 or 1,007 patients. The cutoff value with optimal diagnosis accuracy was 14.6 kPa in 1,007 patients (positive and negative predictive values, 74% and 96%) with discrepancies among the etiological groups. Eighty patients were misclassified: (1) among 45 patients without cirrhosis with liver stiffness 14.6 kPa or greater, 27 (60%) had extensive fibrosis and 10 (22%) significant perisinusoidal fibrosis; and (2) among 35 patients with cirrhosis and liver stiffness less than 14.6 kPa, 10 (29%) had a macronodular pattern and 25 (71%) either none or mild activity. In conclusion, Fibroscan is a reliable method for the diagnosis of cirrhosis in patients with chronic liver diseases, better at excluding than at predicting cirrhosis using a threshold of 14.6 kPa. False negatives are mainly attributable to inactive or macronodular cirrhosis. PMID- 17133504 TI - Lower urinary tract symptoms and bladder control in advanced Parkinson's disease: effects of deep brain stimulation in the subthalamic nucleus. AB - Deep brain stimulation in the subthalamic nucleus (STN) leads to significant improvement in motor function in patients with advanced Parkinson's disease (PD). In this prospective study including 16 patients with PD, we investigated (1) lower urinary tract symptoms (LUTS) by questionnaires International Prostate Symptom Score (IPSS, symptoms only) and Danish Prostate Symptom Score (DanPSS, symptoms and bother of symptoms) and (2) bladder control (assessed by urodynamics) before and after implantation of electrodes in the STN. PD symptoms (Unified Parkinson's Disease Rating Scale score) improved significantly (P < 0.0001), and symptoms of overactive bladder (IPSS) decreased along with the troublesome symptoms of overactive bladder (DanPSS; P < 0.01 for both). Urodynamic parameters before and after implantation of electrodes in the STN, evaluated with and without the stimulation on, did not change significantly. PMID- 17133505 TI - Family-based association study of the restless legs syndrome loci 2 and 3 in a European population. AB - Three loci for the restless legs syndrome (RLS) on chromosomes 12q, 14q, and 9p (RLS1, RLS2, and RLS3) have been mapped, but no gene has been identified as yet. RLS1 has been confirmed in families from three different populations. We conducted a family-based association study of 159 European RLS trios. The subjects were genotyped using microsatellite markers evenly covering the candidate regions on chromosomes 14q and 9p with an average intermarker distance of 1.1 cM. Transmission disequilibrium tests were used to analyze the data, and empirical P values were estimated by permutation testing. On chromosome 14q, a significant association (empirical P = 0.0033) was found with a haplotype formed by markers D14S1014 and D14S1017 when analyzing all families. On chromosome 9p, no significant association in the sample of all families and only marginally significant associations were detected, with a haplotype involving markers D9S1846-D9S171 in a subset of South European trios and with a haplotype at D9S156 D9S157 in a subset of Central European trios (P = 0.0086 and 0.0077, respectively). These results represent the first confirmation of these loci in a mixed European population. Variable results observed in families of different ethnic groups further corroborate the genetic complexity of RLS. PMID- 17133506 TI - Inflation lung mechanics deteriorates markedly after saline instillation and open endotracheal suctioning in mechanically ventilated healthy piglets. AB - INTRODUCTION: Non-bronchoscopic bronchoalveolar lavage is an alternative to diagnostic bronchoscopy in pediatric patients, as fiberoptic bronchoscopes with aspiration channels are too large for small infants. There are many variations of the method in clinical practice, and saline instillation followed by open endotracheal suctioning is still commonly used. Lung function can deteriorate with these procedures, and we have investigated the effects on lung mechanics and oxygenation in healthy piglets. METHODS: The lungs of anesthetized and mechanically ventilated piglets were recruited with CPAP 35 cmH2O. Thereafter we instilled 5 ml of saline into the endotracheal tube, followed by three breaths from the ventilator. Saline was retrieved through a suction catheter wedged far distally in the airway. The procedure was followed by a new recruitment maneuver. Complete inspiratory/expiratory pressure - volume loops (PV-loops) were obtained just before and 5 min after saline instillation. Arterial blood gases were collected at equivalent times in 14 similar piglets submitted to exactly the same procedure. RESULTS: The inspiratory limb of the PV-loops changed markedly, as the lower inflection point was displaced towards higher pressures (P=0.004), and hysteresis measured at 15 and 30 cmH2O increased (P=0.004 and P=0.012, respectively). Although PaO2 decreased significantly (P=0.001), values after saline instillation/suctioning were still in the high normal range, that is, 22.2 +/- 2.6 kPa. CONCLUSIONS: Opening pressures of the lungs increase markedly after saline instillation/suctioning in healthy piglets. In this situation, adequate recruitment maneuvers and PEEP might prevent lung collapse and deteriorations in arterial oxygenation. PMID- 17133507 TI - Effects of dimerization of Serratia marcescens endonuclease on water dynamics. AB - The dynamics and structure of Serratia marcescens endonuclease and its neighboring solvent are investigated by molecular dynamics (MD). Comparisons are made with structural and biochemical experiments. The dimer form is physiologic and functions more processively than the monomer. We previously found a channel formed by connected clusters of waters from the active site to the dimer interface. Here, we show that dimerization clearly changes correlations in the water structure and dynamics in the active site not seen in the monomer. Our results indicate that water at the active sites of the dimer is less affected compared with bulk solvent than in the monomer where it has much slower characteristic relaxation times. Given that water is a required participant in the reaction, this gives a clear advantage to dimerization in the absence of an apparent ability to use both active sites simultaneously. PMID- 17133508 TI - Anodic oxidation and hydrothermal treatment of commercially pure titanium surfaces increases expression of bone morphogenetic protein-2 in the adherent macrophage cell line J774A.1. AB - The surface property of commercially pure titanium (cpTi) was improved by forming a thin hydroxyapatite (HA) layer by anodic oxidation and hydrothermal treatment (HA/cpTi). We hypothesize that the adhesion of macrophages to HA/cpTi surfaces is important to the process of osseointegration. This study investigates the effect of adhesion of macrophages to HA/cpTi surfaces on the expression of bone morphogenetic protein-2 (BMP-2). The murine macrophage cell line J774A.1 was cultured on HA/cpTi and polished cpTi (S/cpTi). Macrophage cell adhesion was examined by SEM, 0-72 h following plating onto HA/cpTi and S/cpTi. BMP-2 gene expression was examined by RT-PCR analysis. The level of BMP-2 secreted into the supernatant was measured using an ELISA assay. The extent of macrophage adhesion increased with time on both the HA/cpTi and S/cpTi surfaces, with a" higher degree of spreading observed on HA/cpTi than onS/cpTi surfaces after 24 or 72 h. The ratio of BMP-2 mRNA was higher on HA/cpTi than on S/cpTi after 24 h (0.348 vs. 0, p < 0.05) and 72 h (0.584 vs. 0.189, p < 0.05). After 24 h, secretion of BMP-2 was detected in cultures grown on HA/cpTi, but not on S/cpTi. After 72 h, secretion of BMP-2 was detected in cultures grown on S/cpTi, but the levels were higher in cultures grown on HA/cpTi. These findings show that macrophages have the capacity to adhere to HA/cpTi endosseous implants and provide a source of osteoinductive cytokines that may play a key role in the process of osseointegration. PMID- 17133509 TI - Calcium phosphate bone cements, functionalized with VEGF: release kinetics and biological activity. AB - Calcium phosphate bone cements are of great interest for bone replacement since the nanocrystalline structure allows their remodelling into native bone tissue. A strategy to accelerate vascularization of the implant region is the functionalization with vascular endothelial growth factor (VEGF), which is known to mediate angiogenesis in vivo. In this study, the release of recombinant human VEGF (rhVEGF(165)) following physical adsorption to Biocement D (BioD) and several modifications were investigated. Our data demonstrate a high VEGF binding capacity of BioD and a sustained release with a moderate initial burst. A proliferation assay using endothelial cells revealed maintenance of biological activity of VEGF after release from BioD. Release behavior of BioD was not improved by modification with mineralized collagen type I, as well as with a combination of mineralized collagen with O-phospho-L-serine and sodium citrate, respectively. In contrast, a positive impact of these modifications on the activity of released VEGF was observed; in case of the phosphoserine- and sodium citrate-modified cements, the biological efficacy of released VEGF was even higher than that of nonreleased control VEGF. We conclude that the bone implant material BioD and, especially, the phosphoserine modification may support activation of angiogenesis by delivery of VEGF in a local and sustained manner. PMID- 17133510 TI - Sporadic geniospasm (chin trembling): report of a case. PMID- 17133511 TI - Fatigue in Parkinson's disease: a review. AB - Fatigue is a common problem in Parkinson's disease (PD), often the most troubling of all symptoms. It is poorly understood, generally under-recognized, and has no known treatment. This article reviews what is known about the symptom, putting it into the context of fatigue in other disorders, and outlines a program for developing better understanding and therapy. PMID- 17133512 TI - Long-term tolerability of tetrabenazine in the treatment of hyperkinetic movement disorders. AB - We sought to review the long-term tolerability of tetrabenazine (TBZ) and seek determinants of tolerability in the treatment of hyperkinetic movement disorders. A retrospective chart review was performed on patients treated with TBZ between 1997 and 2004. Efficacy of TBZ was assessed by a 1- to 5-point response scale (1 = marked reduction in abnormal movements, 5 = worsening). All adverse events (AEs) were captured according to their relationship with study drug. A total of 448 patients (42% male) were treated for a variety of hyperkinesias, including tardive dyskinesia (n = 149), dystonia (n = 132), chorea (n = 98), tics (n = 92), and myoclonus (n = 19). The mean age at onset of the movement disorder was 43.0 +/- 24.2 years, with TBZ starting at a mean age of 50.0 +/- 22.3 years. Patients remained on treatment for a mean of 2.3 +/- 3.4 years. An efficacy response rating of 1 or 2 was sustained in the majority of patients between the first and last visit. Common AEs included drowsiness (25.0%), Parkinsonism (15.4%), depression (7.6%), and akathisia (7.6%). Comparison of log-likelihood ratios revealed that age was a reliable predictor of Parkinsonism (P < 0.0001). TBZ is a safe and effective drug for the long-term treatment of hyperkinetic movement disorders. PMID- 17133513 TI - Phenotypic variation among brothers with the McLeod neuroacanthocytosis syndrome. AB - McLeod syndrome is an X-linked multisystem disorder affecting red blood cells, the peripheral and central nervous systems, and skeletal and cardiac muscle. No clear correlations of the clinical findings with the genotype of XK mutations have yet been uncovered. Here, we report the clinical features and progression in 10 affected brothers from 4 families with McLeod syndrome. There is significant variation in clinical presentation within families, including in causes of morbidity and mortality. This phenotypic variation, despite shared mutations, suggests the action of disease-modifying factors that may explain some of the difficulties with genotype-phenotype correlation in McLeod syndrome. PMID- 17133514 TI - Pyramidal tract degeneration in multiple system atrophy: the relevance of magnetization transfer imaging. AB - The clinical features of multiple system atrophy (MSA) include four domains: autonomic failure/urinary dysfunction, Parkinsonism, cerebellar ataxia, and corticospinal tract dysfunction. Although the diagnosis of definite MSA requires pathological confirmation, magnetic resonance imaging (MRI) studies have been shown to contribute to the diagnosis of MSA. Although pyramidal tract dysfunction is frequent in MSA patients, signs of pyramidal tract involvement are controversially demonstrated by MRI. We evaluated the pyramidal involvement in 10 patients (7 women) with clinically probable MSA, detecting the presence of spasticity, hyperreflexia, and Babinski sign, as well as demonstrating degeneration of the pyramidal tract and primary motor cortex by MRI in all of them. Our article also discusses key radiological features of this syndrome. In MSA, pyramidal tract involvement seems to be more frequent than previously thought, and the clinicoradiological correlation between pyramidal tract dysfunction and degeneration may contribute to the understanding of the clinical hallmarks of MSA. MRI may also add information regarding the differential diagnosis of this syndrome. PMID- 17133515 TI - Substantia nigra hypoechogenicity: definition and findings in restless legs syndrome. AB - Pathological studies demonstrate a decreased iron content in the substantia nigra (SN) contributing to the pathophysiology of restless legs syndrome (RLS). SN echogenicity as measured by transcranial sonography (TCS) correlates with the SN iron content. The objective of this study was to determine a critical value to define SN hypoechogenicity as a potential marker for RLS. There were 49 RLS patients (39 idiopathic, 10 secondary) and 49 age- and sex-matched controls who underwent TCS by 2 independent and blinded examiners to determine the area of SN echogenicity. We found that SN echogenicity is significantly decreased in RLS patients compared to healthy controls (P < 0.001). SN hypoechogenicity (sum area of SN echogenicity of both sides < 0.2 cm(2)) is more common in idiopathic than in secondary RLS patients. The area under curve for idiopathic RLS versus controls (receiver operating characteristics) is 0.91, specificity is 0.90, and sensitivity is 0.82. TCS provides an interesting additional instrument in the diagnosis of RLS. Therefore, SN hypoechogenicity (SN sum area < 0.2 cm(2)), which is supposed to indicate a decreased SN iron content, is a marker for RLS. Further studies are needed to investigate its significance for the pathophysiology of this frequent movement disorder and possible clinical applications. PMID- 17133516 TI - Mixed alien hand syndrome coexisting with left-sided extinction secondary to a left corpus callosal lesion: a case report. AB - Alien hand syndrome (AHS) is actually two distinct syndromes with distinct clinical and anatomic features, that is, a frontal type and a callosal type. Frontal AHS occurs in the dominant hand; is associated with reflexive grasping, groping, and compulsive manipulation of tools. Callosal AHS is characterized primarily by intermanual conflict. We report a case of right frontal AHS and left callosal AHS (mixed AHS) secondary to ischemic stroke of the left corpus callosum (lesion extending from the genu to splenium) and right corpus callosum (minimal lesion in the splenium) in a 67-year-old male patient who also presented with left-sided tactile extinction. To our knowledge, rare reports have documented mixed AHS coexisting with nondominant side extinction secondary only to unilateral (left) callosal lesion, as in our case. PMID- 17133517 TI - Abnormal responses to repetitive transcranial magnetic stimulation in multiple system atrophy. AB - We studied the response of the motor cortex to brief trains of suprathreshold repetitive transcranial magnetic stimulations (rTMS) in patients with the Parkinson-variant of multiple system atrophy (MSA-P) and compared it to patients with idiopathic Parkinson's disease (PD) and healthy controls. Eight subjects were studied in each group, and patients were matched for disease severity as assessed by Hoehn & Yahr stages. rTMS was delivered at rest and during low-level contractions in trains of 10 stimulations at 5 Hz, and stimulation intensity was set to result in an motor evoked potential (MEP) in the first dorsal interosseus muscle of 0.5 to 1.0 mV. In MSA-P, MEP amplitude at rest was already reduced after the second stimulus and remained so, while it did not change in PD and controls. During contraction, MEP size did not change during the train in any group. The silent period that followed the last stimulus was of similar duration as the first stimulus in MSA-P, but was increased in PD and controls. These findings indicate that abnormal inhibition occurs within the motor cortex in MSA P, despite dopaminergic treatment and indicate differences in cortical dysfunction between MSA-P and PD. We suggest that these abnormalities reflect the motor cortex pathology found in MSA-P. PMID- 17133518 TI - Multiple system atrophy in a patient with the spinocerebellar ataxia 3 gene mutation. AB - The cerebellar variant of multiple system atrophy (MSA-C) has overlapping clinical features with the hereditary spinocerebellar ataxias (SCAs), but can usually be distinguished on a clinical basis. We describe a patient who developed a sporadic, late-onset, rapidly progressive neurodegenerative disorder consistent with MSA-C. Genetic testing, however, showed an abnormal expansion of one allele of the spinocerebellar ataxia 3 (SCA3) gene. The clinical impression of MSA-C was confirmed by identification of numerous alpha-synuclein-containing glial cytoplasmic inclusions on autopsy. These findings suggest that abnormal expansion of the SCA3 gene may be a risk factor for the development of MSA-C. PMID- 17133519 TI - Levodopa-induced hyperactivity in mice treated with 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine. AB - The present study examines the motor responses of 10- to 12-month-old, male C57 mice that were either given intraperitoneal (IP) injections of 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP; 30 mg/kg per day) or vehicle for 10 consecutive days, followed by IP injections of levodopa (200 mg/kg) plus carbidopa (25 mg/kg). Five days of MPTP exposure resulted in the Straub tail phenomenon and pronounced hypokinesia. However, during the next 5 days, motor activity returned to baseline, even with continued MPTP treatment. After 10 to 14 days of rest, all mice were then treated with levodopa/carbidopa twice daily for multiple, consecutive days. However, only the previously MPTP-treated animals became hyperkinetic, as compared to levodopa-treated control animals that were not previously exposed to MPTP. Abnormal activity included scratching, running, gnawing, and jumping movements. Hyperactivity lasted for approximately 2 hours after each levodopa injection and then returned to baseline, but the amount of hyperkinesia increased with additional days of levodopa treatment, even though the daily levodopa dose was not changed. These results demonstrate that levodopa can cause reproducible hyperactivity in mice that were previously exposed to MPTP. PMID- 17133520 TI - Brainstem respiratory control: substrates of respiratory failure of multiple system atrophy. AB - Multiple system atrophy may manifest with severe respiratory disorders, including sleep apnea and laryngeal stridor, which reflect a failure of automatic control of respiration. This function depends on a pontomedullary network of interconnected neurons located in the parabrachial/Kolliker Fuse nucleus in the pons, nucleus of the solitary tract, and ventrolateral medulla. Neurons in the preBotzinger complex expressing neurokinin-1 receptors are critically involved in respiratory rhythmogenesis, whereas serotonergic neurons in the medullary raphe and glutamatergic neurons located close to the ventral medullary surface are involved in central chemosensitivity to hypercapnia, hypoxia, or both. Pathological studies using selective neurochemical markers indicate that these neuronal groups are affected in multiple system atrophy. This finding may provide potential anatomical substrates for the respiratory manifestations of the disease. PMID- 17133521 TI - Esophageal stent improves ventilation in a child with a broncho-esophageal fistula caused by Mycobacterium tuberculosis. AB - The deployment of an esophageal stent to aid in the ventilation of a child who had developed an acquired broncho-esophageal fistula caused by Mycobacterium tuberculosis (MTB) is described. The 12-month-old boy presented with respiratory failure requiring ventilation. The air leak via the fistula led to inadequate mechanical ventilation. The deployment of the stent resulted in successful ventilation, closure of the fistula, and eventual successful treatment. PMID- 17133522 TI - Airway pressure release ventilation: a pediatric case series. AB - Airway pressure release ventilation (APRV) is a relatively new mode of mechanical ventilation (MV) first described in animal studies in 1987 and in humans in 1988. It is a time-triggered, time-cycled, pressure-limited mode where a high level of continuous positive airway pressure (CPAP) is maintained with brief regular releases in pressure, and spontaneous breathing is allowed throughout the cycle. In theory, it is consistent with a lung protective approach while having some hypothetical advantages over HFOV. The use of this mode of ventilation in pediatrics has been limited. The authors describe their experience with this mode of ventilation in a series of pediatric patients. PMID- 17133523 TI - Rapid accumulation of nucleostemin in nucleolus during newt regeneration. AB - In newt regeneration, differentiated cells can revert to stem cell-like cells in which the proliferative ability and multipotentiality are restored after dedifferentiation. However, the molecular events that occur during the dedifferentiation still remain obscure. Nucleostemin has been identified in mammals as a nucleolar protein specific to stem cells and cancer cells. In this study, a newt nucleostemin homologue was cloned and its regulation was analyzed. During lens regeneration, the expression of nucleostemin was activated and nucleostemin rapidly accumulated in the nucleoli of dedifferentiating pigmented epithelial cells 2 days before cell cycle reentry. During limb regeneration, nucleostemin also accumulated in the nucleoli of degenerating multinucleate muscle fibers before blastema formation. These findings suggest that nucleostemin plays a role in the dedifferentiation of newt cells and can provide crucial clues for addressing the molecular events at early steps of cellular dedifferentiation in newts. PMID- 17133524 TI - Changing trends in sex specific prevalence rates for childhood asthma, eczema, and hay fever. AB - Numerous surveys of school-aged children have shown increasing asthma prevalence with a less publicized but noticeable change in the male to female ratio. We sought to confirm this change in the sex ratio in four questionnaire-based surveys and investigate possible explanations. Identical questionnaire surveys were performed in 1989 (n=3,390), 1994 (n=4,047), 1999 (n=3,540) and 2004 (n=1,920) in school-children aged 9-11 years. Over these 15 years the male to female ratio (M:F) significantly narrowed for wheeze (1.34 to 0.98:1 P < 0.0002), for asthma (1.74 to 1.02:1 P < 0.0001), for eczema (1.42:1 to 0.81:1 P < 0.0001) and for hay fever (1.46 to 0.93:1 P < 0.0001). The diagnosis of asthma in children with wheeze was more commonly made in boys in 1989 relative risk RR 1.32 (1.12, 1.56), even in those with accompanying eczema and/or hay fever RR 1.20 (0.99, 1.45). By 2004 this sex bias in diagnosis was no longer present, RR 1.01 (0.91, 1.12) for wheeze and 1.02 (0.85, 1.21) for those with wheeze and eczema and/or hay fever. From 1989 to 2004 no significant difference in sex distribution changes between older and younger children occurred, making secular trends in the onset of puberty in females an unlikely contributory factor. The disappearance of the bias to diagnose asthma in symptomatic males but not in females may be partly responsible for the narrowing of the sex ratio, but other factors such as those enhancing the expression of asthma and atopy in females may also be implicated. PMID- 17133525 TI - Invited commentary on the case series by J. Krishnan and W. Morrison: Airway pressure release ventilation: A Pediatric Case Series. Ped Pulmon 2007;42:83-88. PMID- 17133526 TI - Evidence-based analysis of physical therapy in Parkinson's disease with recommendations for practice and research. AB - Physical therapy is often prescribed in Parkinson's disease. To facilitate the uniformity and efficacy of this intervention, we analyzed current evidence and developed practice recommendations. We carried out an evidence-based literature review. The results were supplemented with clinical expertise and patient values and translated into practice recommendations, developed according to international standards for guideline development. A systematic literature search yielded 6 systematic reviews and 23 randomized controlled trials of moderate methodological quality with sufficient data. Six specific core areas for physical therapy were identified: transfers, posture, reaching and grasping, balance, gait, and physical capacity. We extracted four specific treatment recommendations that were based on evidence from more than two controlled trials: cueing strategies to improve gait; cognitive movement strategies to improve transfers; exercises to improve balance; and training of joint mobility and muscle power to improve physical capacity. These practice recommendations provide a basis for current physical therapy in Parkinson's disease in everyday clinical practice, as well as for future research in this field. PMID- 17133527 TI - HMPV infections are frequently accompanied by co-infections. PMID- 17133529 TI - A randomized, sham-controlled, proof of principle study of transcranial direct current stimulation for the treatment of pain in fibromyalgia. AB - OBJECTIVE: Recent evidence suggests that fibromyalgia is a disorder characterized by dysfunctional brain activity. Because transcranial direct current stimulation (tDCS) can modulate brain activity noninvasively and can decrease pain in patients with refractory central pain, we hypothesized that tDCS treatment would result in pain relief in patients with fibromyalgia. METHODS: Thirty-two patients were randomized to receive sham stimulation or real tDCS with the anode centered over the primary motor cortex (M1) or the dorsolateral prefrontal cortex (DLPFC) (2 mA for 20 minutes on 5 consecutive days). A blinded evaluator rated the patient's pain, using the visual analog scale for pain, the clinician's global impression, the patient's global assessment, and the number of tender points. Other symptoms of fibromyalgia were evaluated using the Fibromyalgia Impact Questionnaire and the Short Form 36 Health Survey. Safety was assessed with a battery of neuropsychological tests. To assess potential confounders, we measured mood and anxiety changes throughout the trial. RESULTS: Anodal tDCS of the primary motor cortex induced significantly greater pain improvement compared with sham stimulation and stimulation of the DLPFC (P < 0.0001). Although this effect decreased after treatment ended, it was still significant after 3 weeks of followup (P = 0.004). A small positive impact on quality of life was observed among patients who received anodal M1 stimulation. This treatment was associated with a few mild adverse events, but the frequency of these events in the active treatment groups was similar to that in the sham group. Cognitive changes were similar in all 3 treatment groups. CONCLUSION: Our findings provide initial evidence of a beneficial effect of tDCS in fibromyalgia, thus encouraging further trials. PMID- 17133530 TI - Fear avoidance and prognosis in back pain: a systematic review and synthesis of current evidence. AB - OBJECTIVE: Fear of pain, which is hypothesized to result in avoidance behavior, has been described as an obstacle to recovery in populations of patients with low back pain. However, the evidence to support the link between high levels of fear at early stages of pain and poor prognosis has yet to be systematically assessed. We undertook this review to explore current evidence and to propose further development of theoretical models. METHODS: We performed a systematic literature review of all prospective inception cohorts of patients with acute low back pain that measured fear of pain (often described as fear avoidance) at baseline. RESULTS: We reviewed 9 studies reported between 2001 and 2006. Several of these had acceptable/good methodology. Three studies, of which at least 1 had excellent methodology, showed no link between measures of fear at baseline and poor prognosis in the short term (3 months) or the long term (12 months). Three studies with acceptable methodology showed weak evidence for such a link, but the effect sizes were small. The only study with acceptable methodology to find a clear link suggested that fear of movement was linked to long-term pain. CONCLUSION: Despite the prevalent focus on fear of pain at early stages of back pain, there is little evidence to link such fear states with poor prognosis. There is some evidence to suggest that fear may play a role when pain has become persistent. There is a growing consensus that distress/depression plays an important role at early stages, and clinicians should focus on these factors. PMID- 17133531 TI - Interleukin-10 promoter polymorphisms in giant cell arteritis. AB - OBJECTIVE: To investigate potential associations between interleukin-10 (IL-10) promoter polymorphisms and susceptibility to, and clinical features of, giant cell arteritis (GCA). METHODS: A total of 140 patients with biopsy-proven GCA who were residents of Reggio Emilia, Italy, and 200 population-based controls from the same geographic area were genotyped for promoter polymorphisms of the IL-10 gene, by molecular methods. The patients were subgrouped according to the presence or absence of polymyalgia rheumatica (PMR) and ischemic complications (any or all of the following: vision loss, jaw claudication, cerebrovascular accidents, or aortic arch syndrome). RESULTS: The distribution of the C/A 592 genotype differed significantly between the GCA patients and the controls (P(corr) = 0.003). Carriers of the A592 allele (A/A or C/A) were significantly more frequent among the GCA patients than among the controls (P(corr) = 0.004, odds ratio [OR] 2.0 [95% confidence interval (95% CI) 1.3-3.1]). Homozygosity for the A592 allele was significantly more frequent among the GCA patients than among the controls (P(corr) = 0.002, OR 3.4 [95% CI 1.6-7.2]). The distribution of the A/G 1082 genotype was similar in GCA patients and controls. In the haplotype analysis, the frequency of the ATA haplotype was significantly higher in GCA patients than in the controls (P = 0.0001), whereas the frequencies of the ACC and GTA haplotypes were significantly lower (P = 0.0001 for both comparisons). No significant associations were found for comparisons of GCA patients with and those without PMR or GCA patients with and those without ischemic complications. CONCLUSION: Our findings show that the -592 C/A promoter polymorphism of the IL 10 gene is associated with susceptibility to GCA. PMID- 17133532 TI - Immunohistochemical evidence of a cytokine and chemokine network in three patients with Erdheim-Chester disease: implications for pathogenesis. AB - OBJECTIVE: Erdheim-Chester disease (ECD) is a rare form of non-Langerhans' cell histiocytosis (LCH) of unknown etiology, characterized by diffuse histiocyte infiltration of bones and soft tissue. The purpose of this study was to assess cell proliferation and expression of cytokines, chemokines, and chemokine receptors that may potentially be important in histiocyte accumulation in ECD lesions. METHODS: Biopsies were performed on 3 patients with ECD. The diagnosis of the disease was based on clinical signs including typical radiologic osteosclerosis, and on the detection of foamy CD68+,CD1a- non-Langerhans' cell histiocytes on histologic examination. The expression of the proliferation marker Ki-67 as well as of selected chemokine/chemokine receptor pairs and cytokines was analyzed by immunohistochemistry. RESULTS: In all samples, Ki-67 was undetectable in CD68+ histiocytes. Conversely, these cells expressed the chemokines CCL2 (monocyte chemotactic protein 1), CCL4/macrophage inflammatory protein 1beta (MIP 1beta), CCL5/RANTES, CCL20/MIP-3alpha, and CCL19/MIP-3beta, and their counter receptors CCR1, CCR2, CCR3, CCR5, CCR6, and CCR7. Moreover, ECD histiocytes expressed interferon-gamma-inducible 10-kd protein (CXCL10), which is specifically induced by interferon-gamma, and interleukin-6 and RANKL, which are both implicated in bone remodeling. Finally, all cases showed a Th1-type lymphocyte infiltrate. CONCLUSION: Our data indicate that, similar to LCH, ECD lesions are characterized by a complex cytokine and chemokine network, which may orchestrate histiocyte activation and accumulation through an autocrine loop and contribute to the pathogenesis of the disease. PMID- 17133533 TI - A different type of procedure for a different type of pain. PMID- 17133534 TI - Hematopoietic stem cell transplantation in autoimmune diseases: is the glass half full or half empty? PMID- 17133535 TI - New regulatory rules for clinical trials in the United States and the European Union: key points and comparisons. PMID- 17133536 TI - Lack of genetic association of the interleukin-4 receptor single-nucleotide polymorphisms I50V and Q551R with erosive disease in psoriatic arthritis. PMID- 17133537 TI - Lack of association of antibodies to ribosomal P proteins with lupus membranous glomerulonephritis: comment on the article by Do Nascimento et al. PMID- 17133539 TI - Clinical images: scapular winging. PMID- 17133540 TI - Statistical methods in Arthritis & Rheumatism: current trends. PMID- 17133541 TI - Randomized controlled trials of autologous hematopoietic stem cell transplantation for autoimmune diseases: the evolution from myeloablative to lymphoablative transplant regimens. PMID- 17133543 TI - Presence of significant synovitis in rheumatoid arthritis patients with disease modifying antirheumatic drug-induced clinical remission: evidence from an imaging study may explain structural progression. AB - OBJECTIVE: More timely and effective therapy for rheumatoid arthritis (RA) has contributed to increasing rates of clinical remission. However, progression of structural damage may still occur in patients who have satisfied remission criteria, which suggests that there is ongoing disease activity. This questions the validity of current methods of assessing remission in RA. The purpose of this study was to test the hypothesis that modern joint imaging improves the accuracy of remission measurement in RA. METHODS: We studied 107 RA patients receiving disease-modifying antirheumatic drug therapy who were judged by their consultant rheumatologist to be in remission and 17 normal control subjects. Patients underwent clinical, laboratory, functional, and quality of life assessments. The Disease Activity Score 28-joint assessment and the American College of Rheumatology remission criteria, together with strict clinical definitions of remission, were applied. Imaging of the hands and wrists using standardized acquisition and scoring techniques with conventional 1.5T magnetic resonance imaging (MRI) and ultrasonography (US) were performed. RESULTS: Irrespective of which clinical criteria were applied to determine remission, the majority of patients continued to have evidence of active inflammation, as shown by findings on the imaging assessments. Even in asymptomatic patients with clinically normal joints, MRI showed that 96% had synovitis and 46% had bone marrow edema, and US showed that 73% had gray-scale synovial hypertrophy and 43% had increased power Doppler signal. Only mild synovial thickening was seen in 3 of the control subjects (18%), but no bone marrow edema. CONCLUSION: Most RA patients who satisfied the remission criteria with normal findings on clinical and laboratory studies had imaging-detected synovitis. This subclinical inflammation may explain the observed discrepancy between disease activity and outcome in RA. Imaging assessment may be necessary for the accurate evaluation of disease status and, in particular, for the definition of true remission. PMID- 17133544 TI - Characteristics of diffuse large B cell lymphomas in rheumatoid arthritis. AB - OBJECTIVE: Patients with rheumatoid arthritis (RA) are at increased risk of malignant lymphomas, with a correlation between RA disease severity and lymphoma risk, most pronounced for diffuse large B cell lymphomas (DLBCLs), which also constitute the majority of RA-associated lymphomas. DLBCLs can be further subdivided into germinal center (GC)-like and non-GC-like subtypes, with different cellular origins and prognoses. This study was undertaken to investigate whether RA displays a specific association with any of the DLBCL subtypes. METHODS: We identified 139 patients with DLBCLs within a population based case-control study of 378 RA patients with lymphoma. The DLBCLs were examined for CD10, Bcl-6, and interferon regulatory factor 4 expression patterns, subclassified into GC and non-GC subtypes, and then correlated with clinical parameters. RESULTS: We found a statistically significant predominance of the non GC subtype (97 patients; 70% of all DLBCLs). These patients more often had an advanced stage of lymphoma at diagnosis and had a worse 5-year overall survival rate (16% versus 33%) compared with patients with the GC subtype. There was a strong association with RA disease activity in both subtypes, with >70% of the GC and non-GC cases occurring in RA patients with the highest overall disease activity scores. CONCLUSION: These findings suggest that severe RA is particularly associated with the non-GC subtype of DLBCL, and indicate a critical role of activated peripheral B cells as the cells of origin in these lymphomas. PMID- 17133545 TI - Analysis of the full-length genome of a subgenotype IIIB hepatitis A virus isolate: primers for broadly reactive PCR and genotypic analysis. AB - Among six known subgenotypes (IA, IB, IIA, IIB, IIIA, and IIIB) of human hepatitis A virus (HAV), the complete genomic sequence has not been determined for IIIB. In this study, the full-length genomic sequence of a IIIB HAV isolate (HA-JNG06-90F) recovered from a Japanese patient who contracted sporadic hepatitis A in 1990, was determined. The HA-JNG06-90F genome, which comprised 7462 nt excluding the poly(A) tail, was related most closely to NOR-21 of subgenotype IIIA with an identity of 89.1%, and was only 82.6-83.4% similar to human HAV isolates of genotypes I and II over the entire genome. Comparison of full-length genomic sequences of 20 reported isolates and HA-JNG06-90F generated optimal results for separation of different levels: the nucleotide identities were 80.7-86.6% at the genotype level, 89.1-91.9% at the subgenotype level, and 94.6-99.7% at the isolate level. Similar ranges of nucleotide identity were observed when comparing partial nucleotide sequences of the VP1-2B (481 nt; primer sequences at both ends excluded) and 3C/3D (590 nt) regions, which were amplifiable by PCR with primers designed from well-conserved areas of the HAV genome. All 66 samples with IgM-class HAV antibodies tested positive for HAV RNA by both VP1-2B (481 nt)-PCR and 3C/3D (590 nt)-PCR: subgenotype assignment was concordant in all samples tested (IA [n = 61], IB [n = 1], IIIA [n = 2] and IIIB [n = 2]). These results suggest that two broadly reactive PCRs using primers derived from the VP1-2B and 3C/3D regions, respectively, may be applicable to universal detection and phylogenetic analysis of various HAV strains. PMID- 17133546 TI - A mutation in the interferon sensitivity-determining region is associated with responsiveness to interferon-ribavirin combination therapy in chronic hepatitis patients infected with a Japan-specific subtype of hepatitis C virus genotype 1B. AB - The interferon sensitivity-determining region (ISDR) is useful as a predictive marker of the response to interferon (IFN) therapy for chronic hepatitis patients with a Japan-specific subtype (J-type) of hepatitis C virus (HCV) genotype 1b. This marker is not useful for predicting responsiveness of a worldwide subtype (W type) of HCV 1b, which could explain the restricted usefulness of this system only to Japan. In the present study, we examined the predictive value of the ISDR for ribavirin combination therapy. A total of 79 patients with HCV 1b comprising 35 patients with J-type and 44 patients with W-type were treated with IFN in combination with ribavirin for more than 48 weeks. Mutations in the ISDR were detected more frequently often seen in J-type HCV 1b than in W-type; however, the sustained virological response (SVR) rate for the combination therapy was similar between the two subtypes. Multivariate analysis revealed that factors associated with SVR were IFN dose and the number of amino acid substitutions in ISDR but not with subtypes J and W. The correlation between the number of substitutions in ISDR and responses to IFN-ribavirin combination therapy was restricted to patients with J-type HCV 1b. The ISDR is a useful predictive marker for response to IFN-ribavirin combination therapy in J-type HCV. PMID- 17133547 TI - Rapid immunochromatographic test for hantavirus andes contrasted with capture-IgM ELISA for detection of Andes-specific IgM antibodies. AB - Hantavirus is associated with hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). The clinical diagnosis of hantavirus infections has been confirmed routinely by the immunofluorescence antibody assay (IFA) or enzyme-linked immunosorbent assay (ELISA). A rapid and easy diagnostic test for hantavirus infection is required. A new immunochromatographic assay for hantavirus, POC-PUU, useful for the diagnosis of epidemic nephrophaty associated with hantavirus Puumala in Europe, was evaluated in Chile. This test is based on recombinant N-protein of hantavirus Puumala, and cross-reacts with other hantaviruses. Eighty human sera were selected at random from patients from Southern Chile who were suspected with HPS. The hantavirus capture-IgM ELISA was compared with a commercially available POC-PUU test (POC PUUMALA, Reagena Ltd., Toivala, Finland). The test sensitivity and specificity of the POC-PUU test were 97 and 90%, respectively. It is important to note that although the test is not specific for Andes virus the sensitivity and specificity were above 90%, which indicates good reactivity to the Puumala nucleoprotein antigen. As this test is cost-effective, with a high negative value, rapid and easy to carry out, specialized personnel are not necessary, nor does it require specialized equipment. Its usefulness for diagnosis is important in hospitals far from reference centers and areas with a high incidence of HPS cases. PMID- 17133548 TI - The prevalence of chromosomally integrated human herpesvirus 6 genomes in the blood of UK blood donors. AB - A lesser-recognized form of human herpesvirus 6 (HHV-6) persistence is integration of the viral genome in a host chromosome and high viral copy numbers in blood or sera are characteristic of this phenomenon. A cross-sectional study was performed to determine the frequency of high HHV-6 viral loads in whole blood (>6 log(10) copies/ml) in a population of blood donors in London, UK. Blood samples from 500 anonymized blood donors were collected from one donation center, DNA extracted, and quantitative realtime PCR used to measure viral load. Four samples (0.8%) were found to have high viral copy numbers of HHV-6 (median 6.7 log(10) copies/ml; range 6.5- 6.9 log(10) copies/ml). Cellular DNA was also quantitated using qRT-PCR for beta-globin. By comparing these two results, we calculated that there were between two and five copies of HHV-6 present per cell in these four donors. The median viral load detected in plasma from the four individuals was 3.8 log(10) copies/ml (range 3.5-4.0 log(10) copies/ml). All samples were HHV-6 variant B. In addition, a retrospective analysis of all diagnostic blood samples performed for HHV-6 in our center showed a prevalence of 2.9% of high viral loads characteristic of integration. In conclusion, high viral copy numbers of HHV-6, representing a population of viral integration, is detected in 0.8% of UK blood donors. The presence of high HHV-6 viral loads in healthy normal individuals reiterates the need to consider the confounding effect of HHV-6 viral integration in any laboratory diagnosis of HHV-6 infection. PMID- 17133550 TI - Human T-cell lymphotropic virus types I and II (HTLV-I and -II) infection among seroindeterminate cases in Argentina. AB - Human T-cell lymphotropic virus (HTLV) seroindeterminate cases have been reported among blood donors (BD) and in at-risk populations worldwide, including Argentina. The objective of the present work was to study the presence of HTLV I/II infection and its association to specific Western blot (WB) patterns among healthy BD and at-risk populations in Argentina. We analyzed 83 HTLV-I/II seroindeterminate WB cases diagnosed among BD (n = 49) and in different at-risk populations (n = 34) for human retroviruses infections. Multiple indeterminate WB patterns were observed. Out of the total, 13.2% (11/83) of the cases were found to be HTLV-I/II positive by nested-PCR (n-PCR), including 13.2% (11/83) HTLV-I and 2.4% (2/83) presenting HTLV-I and -II co-infection. Most of their serological profiles showed reactivity to gag or env codified proteins. Two samples amplified only one of the six analyzed genes (1 HTLV-I pol gene and 1 HTLV-II tax gene). There was no association between the presence of Trypanosoma cruzi infection and an HTLV-I/II indeterminate WB pattern (only 3 of the 83 samples were positive for T. cruzi antibodies). In conclusion, the majority of HTLV-seroindeterminate WB donors lacked HTLV provirus and was thus considered uninfected. However, when seroreactivity to Env and Gag proteins are observed on the WB and especially in at-risk populations, HTLV infection should be suspected; such individuals should be followed-up and retested. PMID- 17133549 TI - Human herpesvirus 8 genoprevalence in populations at disparate risks of Kaposi's sarcoma. AB - The prevalence of human herpesvirus 8 (HHV-8) in populations at different risks of developing Kaposi's sarcoma (KS) was assessed using a protocol involving immunomagnetic fractionation of CD45+ blood cells prior to detection of the HHV-8 genome by nested PCR. Preliminary studies using blood of eight gay men infected by human immunodeficiency virus-1 (HIV-1) revealed that, for the detection of HHV 8 DNA derived from open reading frame (ORF) 26 of the HHV-8 genome, this protocol provided substantially higher rates (100%) compared to one involving red blood cell (RBC) lysis (0%) and to another requiring double density gradient centrifugation (DDGC) of leukocytes (13%). When the CD45+ fractionation protocol was applied to samples from 103 other HIV-1-infected patients (the vast majority of whom were gay men) and 100 blood donors, the ORF 26 DNA detection rates obtained were 37% and 8%, respectively. When DNA from the variable region 1 of ORF K1 was additionally amplified from samples of the blood donors, a detection rate of 9% was achieved. This rate was highly concordant with the ORF 26 DNA detection rate. Furthermore, the ORF K1 sequences were predominantly unique, assignable to genotypes A1, A4, and C3. When assays for anti-HHV-8 and anti herpes simplex viruses (HSV) 1 and 2 were applied, significant concordances between HHV-8 DNA detection rates and those relating to anti-HHV-8 and to anti HSV 1 and 2 were more frequently observed for HIV-1-infected patients than for blood donors. The higher-than-expected HHV-8 genoprevalence rate in blood donors requires further confirmation in view of its implications for post-transfusion HHV-8 transmission. PMID- 17133551 TI - Norovirus capsid protein expressed in yeast forms virus-like particles and stimulates systemic and mucosal immunity in mice following an oral administration of raw yeast extracts. AB - Norovirus (NV) gastroenteritis is a widespread disease affecting people of all ages worldwide. A simple, safe, and easily deliverable vaccine may be the key for the control and prevention of NV gastroenteritis. In this study, we demonstrated that a NV recombinant capsid protein (strain VA387, genogroup II.4) expressed in yeast (Pichia pastoris) spontaneously formed virus-like particles (VLPs) like those expressed in other in vitro systems. Oral administration of raw material from the yeast cell lysates containing 0.1 mg of VLPs without an adjuvant resulted in systemic and mucosal immune responses in mice. Significantly higher and earlier responses were observed in mice receiving a higher dose (1 mg per dose) of the antigen. Both the serum and fecal antibodies blocked VA387 VLP binding to its histo-blood group antigen receptors. The animals did not reveal any side effect following the administration of the yeast lysates. Our results indicated that yeast is a simple, effective alternative for NV VLP production. The mice immunization study also indicated that the oral administration of raw yeast extracts without an adjuvant is a safe and simple way which is worth to be studied for vaccine delivery in humans. PMID- 17133552 TI - Seroprevalence of rubella infection after national immunization program in Taiwan: vaccination status and immigration impact. AB - Rubella vaccination was implemented nationwide in 1992 in Taiwan. A cross sectional survey was conducted to determine the age-specific seroprevalence of anti-rubella in female students aged 7-22 years old and women of reproductive age in Taipei County. Seropositivity of anti-rubella antibodies was defined as a serum IgG level > or =10 IU/ml tested by enzyme immunoassay. Information on possible predictors of anti-rubella seronegativity was obtained from structured questionnaire interview. Multivariate-adjusted odds ratios (ORm) with their 95% confidence interval (CI) were derived for each predictor using multiple logistic regression analysis. A total of 826 female students and 318 women were recruited. Anti-rubella seropositive rates were 98.3% (282/287), 99.6% (234/235), 96.8% (179/185), 92.4% (110/119), 81.4% (197/242), and 89.5% (68/76) for the age groups of 7-9, 11-13, 15-17, 19-22, 25-33, and 34-44 years old, respectively. Among female students, negative serology for rubella antibodies was associated significantly with the age group and foreign nationality of mother, showing ORm (95% CI) of 1.2 (1.06-1.27) for each year increase in age, and 20.9 (6.31-68.97), respectively. Among women at the reproductive age, low maternal education level 91.6 (9.12-920.74), unmarried status 21.2 (6.16-72.89), and no rubella vaccination 98.9 (11.64-840.25) were associated significantly with an increased evidence of seronegativity to rubella. The National Rubella Vaccination Program has led to herd immunity of school girls. However, in order to eradicate the Rubella syndrome, greater effort is required to vaccinate foreign brides in Taiwan. PMID- 17133553 TI - Progression towards AIDS leads to increased Torque teno virus and Torque teno minivirus titers in tissues of HIV infected individuals. AB - Torque teno virus (TTV) and Torque teno minivirus (TTMV) are highly prevalent in the general population and although no disease has been associated with these viruses yet, co-infections with other pathological viruses are frequent. Both viruses are extremely heterogeneous, especially for DNA viruses, and the role of the immune system in controlling the infections has yet to be established. In this study the TTV/TTMV viral loads in HIV positive tissues have been investigated for the first time. The titers of both TTV and TTMV were compared in the bone marrow and spleen tissues from three groups: HIV negative individuals, HIV positive individuals and HIV positive individuals who had progressed to AIDS, leading to immunosuppression. Limiting dilution PCR using primers situated in the UTR region of the genome were used to semi-quantitate the virus, and TTV and TTMV were differentiated using melting curve analysis of the PCR product. The AIDS group had significantly higher titers compared with both the HIV positive and negative groups for both bone marrow (AIDS vs. HIV positive P = 0.006, AIDS vs. HIV negative P < 0.001) and spleen (AIDS vs. HIV positive P = 0.022, AIDS vs. HIV negative P < 0.001). Analysis of TTV/TTMV titer with CD4 T lymphocyte count showed a significant inverse correlation however neither HCV co-infection or type of Anellovirus infection (single TTV or TTMV, or mixed TTV/TTMV) showed any significant correlation with virus titer. The results show a link between deterioration of the immune system and increased the viral loads in studied tissues. PMID- 17133554 TI - Comparison of hepatitis A and E virus infections among healthy children in Mongolia: evidence for infection with a subgenotype IA HAV in children. AB - To compare the epidemiologic profiles of hepatitis A virus (HAV) and hepatitis E virus (HEV) infections in children in Mongolia, the prevalence of HAV and HEV infections was investigated serologically and molecularly among 717 apparently healthy individuals of 0-20 years of age (mean +/- standard deviation, 8.6 +/- 4.9 years) using serum samples obtained between October 2005 and January 2006. Total antibody against HAV (anti-HAV [total]) was detected in 494 (68.9%) of the 717 subjects, while IgG antibody against HEV (anti-HEV IgG) was detected in only five subjects (0.7%) (P < 0.0001). All five subjects who had anti-HEV IgG, were negative for anti-HEV IgM and HEV RNA. Anti-HAV was detectable in 24 (75.0%) of the 32 infants aged 7 days to 6 months, but not in any of the 8 infants aged 7 to <12 months. The prevalence of anti-HAV was 19.5% (17/87) in the age group of 1-3 years, and it increased to 50.0% (69/138) in the age group of 4-6 years, and further to 81.4% (105/129) in the age group of 7-9 years. Of note, 97.2% of the subjects in the age group of 16-20 years had anti-HAV. The presence of HAV RNA was tested in all 717 subjects, and three children of 1, 4, or 8 years of age were found to have detectable HAV RNA (subgenotype IA). No subject had a history of hepatitis or jaundice. In conclusion, HEV infection was uncommon, but HAV infection lacking overt clinical features was prevalent among children in Mongolia. PMID- 17133555 TI - Genetic diversity of HCV genotype 2 strains in south western France. AB - Hepatitis C virus (HCV) is a major cause of chronic hepatitis and liver disease worldwide. The genetic heterogeneity of HCV and its spread among infected patients can be examined accurately by nucleotide sequencing. The diversity of HCV genotype 2 strains (HCV-2) was studied in a large cohort of patients in the Midi Pyrenees area of southern France. Phylogenetic analysis was performed on 344 NS5B sequences from patients infected with HCV-2. These included 145 strains whose E2 region was also analyzed, and epidemiological data were collected for the corresponding patients. HCV-2 accounts for 11.3% of HCV infections in this area. Phylogenetic analysis of NS5B sequences revealed eight subtypes, while that of the E2 region provided congruent results for 100% of strains. The most frequent subtypes were 2i (24.7%), 2k (22.4%) 2c (17.4%), and 2a (10.8%). The mean age of HCV-2-infected patients was 55.5 years. Epidemiological data showed that blood transfusion is the major route of infection, but it was not associated with any particular subtype. By contrast, intravenous drug users were infected predominantly with genotype 2a. HCV-2a-infected patients were younger than patients infected with other subtypes (48 vs. 56.5 years, P < 0.01). This study shows substantial genetic diversity of HCV-2 subtypes in the south of France and the spread of 2a strains via intravenous drug users. PMID- 17133556 TI - Differences in replication capacity between enterovirus 71 isolates obtained from patients with encephalitis and those obtained from patients with herpangina in Taiwan. AB - The cellular-tropism and biological characteristics of enterovirus 71 (EV71) isolates in Taiwan (TW) were studied. Growth curve experiments were conducted using cell lines that were possibly exhibited pathogenesis, and RT-PCR and sequencing tests were undertaken to amplify the 5' non-coding region (5'-NCR). The encephalitis isolate EV71 TW98NTU2078 was PBMC-tropic, temperature-resistant (Tr) at 40 degrees C, and easier to replicate in HTB-14 (astrocytoma) than the herpangina isolate EV71 TW98NTU1186 (The viral yields were 100-fold higher than those of the herpangina isolate EV71 TW98NTU1186 at 96 hr post infection.). The herpangina isolate EV71 TW98NTU1186 was non-PBMC-tropic, and temperature sensitive (Ts) at 40 degrees C. The replication of EV71 TW98NTU1186 in HTB-14 was lower. No EV71 isolate infected HTB-37 (human colon adenocarcinoma cells). The encephalitis EV71 isolate exhibited better replication and transmission in PBMCs and astrocytes than did the EV71 isolate without CNS involvement. PMID- 17133557 TI - Molecular epidemiology of norovirus gastroenteritis outbreaks in North Carolina, United States: 1995-2000. AB - Noroviruses (NoVs) are the most common cause of acute non-bacterial gastroenteritis outbreaks in the US. We investigated 16 gastroenteritis outbreaks in North Carolina (NC), from 1995 to 2000, to further characterize the epidemiology of NoV using RT-PCR on stool and ELISA on sera. NoV were identified in 14 outbreaks by RT-PCR. Sequence analyses of the amplicons indicated the outbreak strains belonged to the following clusters: five GII/4, three GI/3, one GI/4, one GII/2, one GII/5, one GII/7, and one GII/13 (prototype strain). We detected NoV in stool samples from one outbreak but could not determine its specific cluster within the GII genogroup based on polymerase sequence analysis. The five GII/4 strains were classified as the "95/96 US common strain" and occurred throughout the 5-year period. In contrast to national trends, the majority (86%) of NoV outbreaks identified in North Carolina were foodborne. Of the 12 food-related NoV outbreaks, we were able to document transmission by food handlers in two outbreaks. Person-to-person transmission from primary cases was suggested in three outbreaks. Our results indicate that NoVs are important agents of viral gastroenteritis outbreaks in NC. PMID- 17133558 TI - Identification of NV-F virus DNA in hepatocellular carcinoma. AB - A fragment of DNA sequence derived from a hepatotropic virus, named NV-F was isolated recently. The aim of this study was to examine whether this virus was associated with hepatocellular carcinoma (HCC). Total cellular DNA was extracted from hepatocellular carcinoma tissues. NV-F virus DNA was detected by PCR. The PCR products were subjected to sequence analysis. Of the 78 HCC samples included, 12 (15.4%) were positive for NV-F virus DNA. Sequence analysis of the 12 amplified DNA fragments revealed a point mutation in one of them. The clinicopathological parameters between patients with and without NV-F virus infection were compared. It was found that patients with NV-F virus infection were older than those without NV-F virus infection (mean ages, 61.5 versus 52.5 years; P = 0.032). Otherwise, no difference was observed between the two groups. Of the 12 HCC patients positive for NV-F virus DNA, 11 patients were co-infected by either hepatitis B or C virus. The remaining patient was a Taiwanese aboriginal inhabitant with cryptogenic cirrhosis. In conclusion, NV-F virus DNA was identified in 15.4% of HCC tissues. HCC patients with NV-F virus infection were significantly older than those without NV-F virus infection. PMID- 17133559 TI - Individualized monitoring of drug bioavailability and immunogenicity in rheumatoid arthritis patients treated with the tumor necrosis factor alpha inhibitor infliximab. AB - OBJECTIVE: Infliximab, an anti-tumor necrosis factor alpha (anti-TNFalpha) antibody, is effective in the treatment of several immunoinflammatory diseases. However, many patients experience primary or secondary response failure, suggesting that individualization of treatment regimens may be beneficial. This study was undertaken to investigate whether serologic monitoring of infliximab bioavailability and immunogenicity in individual patients would be useful in optimizing treatment regimens to improve efficacy and tolerability. METHODS: To avoid the use of solid-phase assays, two radioimmunoassays were developed: one for measurement of levels of anti-infliximab antibody, and a functional one for measurement of TNFalpha binding due to infliximab. Sera from 106 randomly selected rheumatoid arthritis patients were tested within 6 months of therapy initiation, and associations between findings of serum assays and disease activity, infusion reactions, and treatment failure occurring within 18 months were assessed. RESULTS: Trough serum infliximab levels after the first 2 intravenous infusions of infliximab at 3 mg/kg varied considerably between patients (range 0-22 microg/ml). At this stage, only 13% of the patients were anti-infliximab antibody positive. With subsequent infusions, the frequency of antibody positivity rose to 30% and 44% (at 3 months and 6 months, respectively), accompanied by diminished trough levels of infliximab. Indeed, low infliximab levels at 1.5 months predicted antibody development and later treatment failure. There were highly significant correlations between high levels of antibodies and later dose increases, side effects, and cessation of therapy. High baseline disease activity, judged by C-reactive protein level and Disease Activity Score, was associated with low levels of infliximab at the early stage of treatment and later development of anti-infliximab antibodies. Cotreatment with methotrexate resulted in slightly reduced antibody levels after 6 months; other disease modifying antirheumatic drugs and prednisolone had no effect. CONCLUSION: Development of anti-infliximab antibodies, heralded by low preinfusion serum infliximab levels, is associated with increased risk of infusion reaction and treatment failure. Early monitoring may help optimize dosing regimens for individual patients, diminish side effects, and prevent prolonged use of inadequate infliximab therapy. PMID- 17133560 TI - Isotype distribution of anti-cyclic citrullinated peptide antibodies in undifferentiated arthritis and rheumatoid arthritis reflects an ongoing immune response. AB - OBJECTIVE: The evolution of the rheumatoid arthritis (RA)-specific anti-cyclic citrullinated peptide (anti-CCP) antibody response, as measured by the isotypes of anti-CCP, has not been described. This study was undertaken to determine anti CCP isotype usage in patients with undifferentiated arthritis (UA), patients with recent-onset RA, and patients with RA of long duration. METHODS: IgA, IgM, and IgG subclasses of anti-CCP were measured by enzyme-linked immunosorbent assay in serum samples that were obtained from IgG anti-CCP antibody-positive patients with UA (n = 110) and IgG anti-CCP antibody-positive patients with RA (n = 152) early after the onset of arthritis. Patients with UA in whom RA developed within 1 year (UA-->RA) were compared with patients with UA in whom RA did not develop within 1 year (UA-->UA). In addition, baseline serum samples obtained from a subset of patients with RA (n = 64) were compared with sera obtained from the same patients a median of 7 years later. RESULTS: IgM anti-CCP was present in early samples from both patients with UA and patients with RA and in followup samples from patients with RA. Several IgG anti-CCP antibody-positive patients who did not have IgM anti-CCP early after disease onset did display IgM anti-CCP later in the course of the arthritis. A diverse pattern of isotype usage was detected in early samples, with a trend toward lower frequencies of all isotypes of anti-CCP in patients with UA compared with patients with RA and in UA-->UA patients compared with UA-->RA patients. Levels of all isotypes except IgG1 had decreased after 7 years. CONCLUSION: These data indicate development of the anti CCP isotype repertoire into full usage early in the course of arthritis. The sustained presence of IgM anti-CCP indicates ongoing recruitment of new B cells into the anti-CCP response, reflecting a continuous (re)activation of the RA specific anti-CCP response during the course of anti-CCP-positive arthritis. PMID- 17133561 TI - Phenomenology of dreams in Parkinson's disease. AB - Rapid eye movement sleep behavior disorder (RBD) occurs in approximately one third of patients with Parkinson's disease (PD) and is associated with a loss of muscle atonia during REM sleep and aggressive dream content. We examined the dream characteristics of PD patients to determine whether dream content differed between patients with RBD and without RBD, men and women with RBD, and men and women with PD. One hundred-twenty patients with a diagnosis of idiopathic PD were consecutively recruited from a movement disorders clinic and were assessed for RBD using clinical diagnostic criteria of the International Classification of Sleep Disorders Revised (2001). Verbatim dream content was obtained from each patient and categorized into dream themes that were coded into nominal categories. Fisher's exact tests determined whether particular dreams were correlated with RBD versus non-RBD, men and women with RBD, and men and women with PD. RBD patients had a higher percentage of violent dreams compared to non RBD patients. There were no significant sex differences in the dream content of RBD patients. Men with PD had more aggressive dreams compared to females with PD. Aggressive dream content was characteristic of RBD patients and sex differences exist in the dream content of the PD population. PMID- 17133562 TI - The role of oxidative stress and antioxidant treatment in liver surgery and transplantation. PMID- 17133563 TI - Functional hepatic imaging and disease recurrence after liver transplantation. PMID- 17133564 TI - Mycophenolate mofetil in combination with reduction of calcineurin inhibitors for chronic renal dysfunction after liver transplantation. AB - The purpose of the study was to introduce mycophenolate mofetil (MMF) in liver transplant recipients with renal dysfunction to decrease calcineurin inhibitor (CNI) dosages without increasing rejection risk. In this prospective, multicenter, randomized study, chronic CNI-related renal dysfunction was defined by an increase in serum creatinine with values >140 micromol/L and <300 micromol/L. Patients were randomized in 2 groups. STUDY GROUP: combination of MMF (2 to 3 g/day) and reduced dose of CNI >or=50% of initial dose; control group: no MMF, but with the ability to reduce CNI doses, but not below 75% of initial dose. Fifty-six patients were included, 27 in the study group and 29 in the control group. In the study group, there was a significant decrease in serum creatinine values, from 171.7 +/- 24.2 micromol/L at day 0 to 143.4 +/- 19 micromol/L at month 12 and a significant increase in creatinine clearance, from 42.6 +/- 10.9 mL/min to 51.7 +/- 13.8 mL/min. No rejection episode was observed in the study group. In the control group, there was no improvement of renal function, assessed by the changes in serum creatinine values, from 175.4 +/- 23.4 micromol/L at day 0 to 181.6 +/- 63 micromol/L at month 12, and in creatinine clearance, from 42.8 +/- 12.8 mL/min to 44.8 +/- 19.7 mL/min. The differences between the 2 groups were significant: P = 0.001 for serum creatinine, and P = 0.04 for creatinine clearance. In conclusion, the introduction of MMF combined with the reduction of at least 50% of CNI dose allowed the renal function of liver transplant recipients to significantly improve at 1 year, without any rejection episode and without significant secondary effects. PMID- 17133565 TI - Alpha-fetoprotein and prognosis in acute liver failure. AB - Serum concentrations of alpha-fetoprotein (AFP), variably elevated during liver injury, have been suggested to be of prognostic importance in acute liver failure (ALF), higher values being associated with improved outcome. Using a nephelometric assay, we measured AFP in sera obtained on admission from 206 patients prospectively enrolled in the US ALF Study, and on day 3 in 162 of these patients. The AFP ratio was defined as the day 3 AFP concentration divided by that observed on day 1. Median (range) admission serum AFP in all patients was 8.1 (1-1,811) ng/mL and increased to 17.6 (1.1-1,162) ng/mL on day 3 (P < 0.001). Higher absolute levels were not associated with improved outcome. In fact, admission AFP levels were lower in survivors not receiving transplants than in those who died or were transplanted (P < 0.001), whereas there was no difference between the 2 groups on day 3 (P = 0.34). However, a rise in AFP values between day 1 and day 3 indicated a better prognosis: the AFP ratio was 2.2 (0.11-22.1) in spontaneous survivors and 0.87 (0.11-16.4) in nonsurvivors (P < 0.001). An increasing AFP level indicated by an AFP ratio >or=1 was observed in 70 of 98 (71%) survivors, whereas a ratio <1 was observed in 51 of 64 (80%) nonsurvivors. In conclusion, AFP values change dynamically during ALF. In this large prospective study, higher absolute values of AFP did not predict a favorable outcome, but a rising level of AFP over the first 3 hospital days frequently indicated survival. PMID- 17133566 TI - Quality of life and psychiatric complications after adult living donor liver transplantation. AB - We investigated the psychosocial effects of a right hepatectomy on donors for adult living donor liver transplantation (ALDLT). Questionnaires were sent to 66 actual donors, who had undergone ALDLT between August 1998 and September 2003, as well as to 139 potential donors, who had been examined as possible candidates for ALDLT; the latter had been excluded and had not undergone surgery. All actual donors reported full recovery within an average period of 14.41 (standard deviation = 8.86) weeks; all had returned to their preoperative employment. In preparation for ALDLT, they had received significantly more support from their families in the decision-making process than the potential donors had (t = 2.02; degree of freedom = 79; P = 0.047); they also felt better informed about donation than the potential donors (t = 2.04; df = 64; P = 0.045). Psychiatric problems occurred in 6 (14%) female donors in the perioperative period, mostly in connection with unrealistic outcome expectations. Donors with severe postoperative complications (n = 3) demonstrated higher scores of psychiatric symptoms (chi-square = 6.39; df = 2; P = 0.041). When we compared potential and actual donors, a significant difference in emotional quality of life was not demonstrated (t = 0.41; df = 76; P = 0.684), and it corresponded to that of the normative sample. For donors, perceived emotional quality of life did not depend on the course of recovery of the recipients. Six to 9 months after donation, potential donors reported a significantly higher physical quality of life than actual donors (t = 2.20; df = 56; P = 0.032). In conclusion, female donors, donors with their own major complications, or donors with unrealistic outcome expectations should be provided with adequate psychosocial care. With regard to the psychosocial outcome, ALDLT is a safe intervention for the donor. PMID- 17133567 TI - Vitamin A toxicity: when one a day doesn't keep the doctor away. AB - Vitamin A toxicity has been reported to cause severe liver disease and, occasionally, liver failure. Herein we present the case of a 60-year-old male with symptoms of muscle soreness, alopecia, nail dystrophy, and ascites. He continued to deteriorate with the development of refractory ascites, renal insufficiency, encephalopathy, and failure to thrive. A liver biopsy demonstrated presence of Ito cells and vacuolated Kupffer cells without the presence of cirrhosis. His clinical history revealed ingestion of large doses of vitamin A. His worsening clinical situation ruled out the possibility of a transjugular intrahepatic portosystemic shunt. The patient underwent orthotopic liver transplantation with resolution of symptoms. Vitamin A toxicity should be considered in the differential diagnosis of noncirrhotic portal hypertension. In conclusion, liver transplantation is a valid option if no improvement occurs in spite of cessation of the medication. PMID- 17133568 TI - Arterioportal fistula requiring liver transplantation. PMID- 17133569 TI - HSV infection and immunosuppression. PMID- 17133571 TI - Liver transplantation for fulminant hepatitis related to nevirapine therapy. AB - Drug-induced hepatotoxicity is an important cause of hepatocellular injury. Non nucleoside retroviral transcriptase inhibitors are known to cause hepatotoxicity. We describe a detailed case of fulminant hepatitis induced by nevirapine (Viramune) and treated by liver transplantation. PMID- 17133572 TI - Conversion to sirolimus: a useful strategy for recalcitrant cutaneous viral warts in liver transplant recipient. AB - Dermatological complications following transplantation are very common and the majority of immunosuppressed transplant recipients develop some to many warts due to human papillomavirus (HPV) infection. In the setting of immunosuppression, therapeutic management may be disappointing because of the extent of the lesions in patients unable to develop a sufficient immune response directed against HPV. We report here a case of a young liver transplant recipient who developed diffuse recalcitrant HPV-induced warts leading to an impairment of her quality of life. Taking into account the antiproliferative and cytostatic properties of the target of-rapamycin (TOR) inhibitors, a new class of immunosuppressive drug, we significantly modified the immunosuppressive regimen. Conversion to sirolimus was followed by a rapid improvement of cutaneous state suggesting that this strategy may be useful for recalcitrant cutaneous viral warts in transplant recipient. PMID- 17133573 TI - Acute liver failure induced by green tea extracts: case report and review of the literature. AB - In industrialized countries, over-the-counter dietary supplements have become popular in preventing and treating an expanding list of medical conditions. Although most commercially available supplements have not been rigorously tested for safety and efficacy, they have found an enlarging market because they are considered natural. Oral supplements containing green tea extract have been marketed as effective for weight loss and to prevent and cure some solid tumors. Although there is little scientific evidence of the effectiveness of green tea extracts to improve the quality of health of regular consumers, there is an increasing body of medical literature supporting the hypothesis that they can cause serious side effects. Our experience adds to previous reports of acute liver toxicity observed in individuals consuming supplements containing green tea extract. We highlight the importance of obtaining a detailed history of dietary supplement consumption when evaluating a patient presenting with acute liver dysfunction. PMID- 17133574 TI - Expanding HCC criteria for liver transplant: the urgent need for prospective, robust data. PMID- 17133575 TI - Relative adrenal insufficiency manifested with multiple organ dysfunction in a liver transplant patient. AB - Relative adrenal insufficiency is now a well-known clinical condition that occurs in critically ill patients particularly with septic complication. However, this pathology has long been unrecognized until recently in liver transplantation patients, for whom postoperative immunosuppressive therapies almost always comprise corticosteroids. We report an obvious case of relative adrenal insufficiency manifested by severe multiple organ dysfunction in a recipient after living donor liver transplantation (LDLT). A 38-year-old woman with multiple hepatocellular carcinoma developed refractory liver failure 2 months after the completion of the dual treatment; namely a cytoreductive right hepatectomy for bulky main tumors followed by 2 courses of percutaneous isolated hepatic perfusion for residual tumors in the remnant liver. She underwent a right lobe LDLT, and postoperative immunosuppression was initiated with a low-dose tacrolimus monotherapy without corticosteroid because of a severe septic condition before transplantation. Postoperatively, she developed progressive hyperbilirubinemia, renal dysfunction, and coagulopathy. As the corticotropin stimulation test suggested the relative adrenal insufficiency, corticosteroid was commenced 40 days after LDLT. Thereafter, multiple organ dysfunction resolved dramatically and promptly. The patient is presently alive and well with completely normalized liver function 45 months after LDLT. PMID- 17133576 TI - Emergency liver transplantation for hereditary lysozyme amyloidosis. PMID- 17133577 TI - Association of citrullinated proteins with synovial exosomes. AB - OBJECTIVE: In addition to releasing proteins and mediators, cells also release membrane vesicles (exosomes and apoptotic blebs) into the extracellular environment. Apoptotic blebs contain multiple autoantigens, but few data are available concerning the protein content of exosomes. Exosomes are formed during an immune response and can directly stimulate T cells or bind to dendritic cells. The aim of this study was to identify the nature of synovial exosomes from patients with different rheumatic diseases and to examine their potential autoantigenic content, which may be involved in the induction of an autoimmune response. METHODS: Synovial exosomes from patients with rheumatoid arthritis (RA), patients with reactive arthritis, and patients with osteoarthritis were purified, analyzed by electron microscopy, and labeled with immunogold to detect IgG and IgM molecules. Autoantigen content was identified by 2-dimensional electrophoresis-immunoblotting and subsequent mass spectrometry. In order to investigate the presence of citrullinated proteins, immunoblotting with anticitrulline antibodies was performed. RESULTS: Citrullinated proteins were observed in all exosome preparations, in contrast to other autoantigenic proteins (e.g., BiP and heterogeneous nuclear RNP A2) that were previously observed in RA and other autoimmune diseases. These citrullinated proteins included the fibrin alpha-chain fragment, fibrin beta-chain, fibrinogen beta-chain precursor, fibrinogen D fragment, and the Sp alpha (CD5 antigen-like protein) receptor. Purification of synovial exosomes led to the detection of citrullinated fibrinogen and citrullinated Sp alpha associated with IgM and IgG. CONCLUSION: Synovial exosomes contain citrullinated proteins, which are known to be autoantigens in RA. Although immune mechanisms in which exosomes carry citrullinated peptides could play an important role in the induction and distribution of citrullinated proteins, there must be a specific recognition of these proteins that is unique to the RA immune system. PMID- 17133578 TI - Analysis of IRF5 gene functional polymorphisms in rheumatoid arthritis. AB - OBJECTIVE: Recent findings suggest that interferon regulatory factor 5 (IRF-5) may play a crucial role in several cellular processes, including the transcription of genes for inflammatory cytokines. Two genetic variants of the IRF5 gene (rs2004640 in exon 1 and rs2280714 in the 3'-untranslated region) have been shown to exert functional modifications affecting IRF5 messenger RNA splicing and expression, and have been associated with genetic predisposition to systemic lupus erythematosus (SLE). The aim of this study was to analyze the possible contribution of the IRF5 gene to the predisposition to rheumatoid arthritis (RA). METHODS: Three case-control cohorts from Spain (724 RA patients and 542 healthy controls), Sweden (281 RA patients 474 healthy controls), and Argentina (284 RA patients and 286 healthy controls) were independently analyzed. Genotyping for IRF5 rs2004640 and rs2280714 was performed using a TaqMan 5' allele-discrimination assay. RESULTS: In the 3 cohorts studied, no statistically significant differences in allele or genotype frequencies of the rs2004640 and rs2280714 IRF5 polymorphisms were observed between RA patients and controls. Accordingly, haplotype analysis revealed that none of the IRF5 haplotypes was associated with genetic predisposition to RA. CONCLUSION: Our results suggest that the IRF5 functional polymorphisms analyzed do not seem to be implicated in genetic susceptibility to RA. PMID- 17133579 TI - Rheumatoid arthritis association with the FCRL3 -169C polymorphism is restricted to PTPN22 1858T-homozygous individuals in a Canadian population. AB - OBJECTIVE: Variants in genes encoding the Fc receptor-like 3 (FcRL-3) and the class II major histocompatibility complex (MHC) transactivator proteins have been associated with an increased risk of rheumatoid arthritis (RA) in Japanese and Nordic populations, respectively. The aim of this study was to investigate these associations in a Canadian Caucasian cohort of RA cases and healthy controls. METHODS: A total of 1,187 RA patients and 462 healthy controls were genotyped for FCRL3 and MHC2TA gene variants associated with RA. Epistasis between the FCRL3 169C and the PTPN22 1858T variants was also examined. RESULTS: An association was detected between RA and both the FCRL3 -169C allele (OR 1.19, P = 0.023) and the homozygous genotype (OR 1.41, P = 0.027), but association of the MHC2TA promoter region variant (-168G) with RA was not replicated. Stratification of the RA cohort by PTPN22 genotypes revealed the FCRL3 risk variant and RA association was stronger in the patient subgroup lacking PTPN22 1858T variants (P = 0.004) and was not detectable in the subgroup with PTPN22 1858T variants (P = 0.52). The PTPN22 association with RA was greater in the absence than in the presence of the FCRL3 -169C allele (P = 0.0008 versus P = 0.001). The PTPN22 1858T variant also increased the risk of autoimmune thyroid disease (AITD) in the RA patients, whereas the FCRL3 risk variant was protective against AITD. CONCLUSION: Our findings support an association of RA with an FCRL3 functional polymorphism and reveal that this association is stronger in the absence of PTPN22 risk genotypes. These findings support a genetic heterogeneity across RA populations, suggesting that both the FCRL3 and PTPN22 genes play roles in RA susceptibility, but in different individuals. PMID- 17133580 TI - The functional variant of the inhibitory Fcgamma receptor IIb (CD32B) is associated with the rate of radiologic joint damage and dendritic cell function in rheumatoid arthritis. AB - OBJECTIVE: Fcgamma receptors (FcgammaRs) recognize immune complexes (ICs) and coordinate the immune response by modulating the functions of dendritic cells (DCs). The purpose of this study was to unravel the role of the inhibitory FcgammaRIIb in rheumatoid arthritis (RA) by studying the effect of the FCGR2B 695T>C polymorphism on susceptibility to RA, severity of the disease, and DC function. METHODS: Genotyping was performed in RA patients (n = 246) and healthy blood donors (n = 269). The patients' demographic data, disease severity, and disease progression were assessed over a followup of 6 years. DCs were cultured for flow cytometry to determine the expression of FcgammaRs. For detection of FcgammaRIIb (CD32B), a unique anti-FcgammaRIIb antibody (2B6-fluorescein isothiocyanate [FITC]) was used. The capacity for antigen uptake by DCs was studied by assessing the uptake of FITC-labeled ICs. Levels of cytokine production by DCs were measured during lipopolysaccharide-mediated cell activation in the presence and absence of ICs. RESULTS: Although no role of the FCGR2B variant in RA susceptibility was demonstrated, this variant was associated with a nearly doubled rate of radiologic joint damage during the first 6 years of RA. Multiple regression analysis showed that FCGR2B was by far the strongest predictor of joint damage identified to date. DCs from patients carrying this variant failed to display the inhibitory phenotype normally observed upon IC mediated triggering of inflammation and displayed diminished FcgammaRII-mediated antigen uptake compared with wild-type DCs. However, the levels of FcgammaRs were not affected, suggesting that the FCGR2B variant alters the function rather than regulation of proteins. CONCLUSION: This study is the first to show that a single genetic variant, the FCGR2B 695T>C polymorphism, is a critical determinant of disease severity in RA and radically changes DC behavior. Our results underscore the key role of DCs in the progression of RA and reveal FcgammaRIIb as an important potential therapeutic target in RA and other autoimmune conditions. PMID- 17133581 TI - The -169C/T polymorphism in FCRL3 is not associated with susceptibility to rheumatoid arthritis or systemic lupus erythematosus in a case-control study of Koreans. AB - OBJECTIVE: In Japanese individuals, the -169C/T single-nucleotide polymorphism (SNP) in FCRL3 has been reported to be associated with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and autoimmune thyroid diseases. The objective of this study was to test the association of this SNP with RA and SLE, in a case-control study of Korean individuals. METHODS: The -169C/T SNP in FCRL3 was genotyped in 1,060 patients with RA, 457 patients with SLE, and 697 unaffected control subjects, using the MassARRAY SNP genotyping system. Associations were tested by multivariate logistic regression, with adjustments for age and sex. RESULTS: No association was detected between the -169C/T SNP and RA (odds ratio [OR] 1.11, 95% confidence interval [95% CI] 0.83-1.48, P = 0.50) or SLE (OR 1.00, 95% CI 0.73-1.37, P = 0.99). This SNP was not associated with rheumatoid factor status, shared epitope status, radiographic severity in patients with RA, or disease manifestations in patients with SLE. CONCLUSION: The association of the -169C/T SNP in FCRL3 with RA and SLE that was observed in Japanese patients was not replicated in a Korean population. PMID- 17133582 TI - Efficacy of pramipexole in restless legs syndrome: a six-week, multicenter, randomized, double-blind study (effect-RLS study). AB - We evaluated the efficacy of pramipexole versus placebo in restless legs syndrome (RLS) for 6 weeks. Overall, 345 patients were randomly assigned in a 1:2 ratio to receive either placebo (n = 115) or pramipexole (n = 230) with a starting dose of 0.125 mg/day. The dose was individually optimized according to the Patient Global Impression (PGI) assessment, up to a maximum of 0.75 mg/day. The primary endpoint consisted of two assessments: the change from baseline in the International RLS Study Group Rating Scale (IRLS) and the proportion of patients with Clinical Global Impressions-Improvement (CGI-I) assessments of "much/very much improved" (CGI-I responders) at week 6. Secondary endpoints included PGI and IRLS responder rates. Patient demographics and baseline characteristics were comparable between treatment groups. At baseline, mean IRLS scores were 24.9 (placebo) and 24.7 (pramipexole), representing severely affected patients. After 6 weeks, adjusted mean reductions (+/-SE) in IRLS score were 5.7 (+/-0.9) for placebo (median dose 0.47 mg/day) and 12.3 (+/-0.6) for pramipexole (median dose 0.35 mg/day; P < 0.0001). CGI-I responder rates were 32.5% (placebo) and 62.9% (pramipexole) (P < 0.0001). For all secondary endpoints, pramipexole showed superior results. Pramipexole was well tolerated throughout the study. PMID- 17133583 TI - Physiological incompatibilities of porcine hepatocytes for clinical liver support. AB - In fulminant hepatic failure, the use of bioartificial liver support (BAL) with porcine hepatocytes is the subject of a current and controversial debate.1 Specifically, the issue of cross-species physiological incompatibilities has not been addressed so far. We therefore investigated the effects of species-specific cytokines in single and cocultures on hepatocyte function. Hepatocyte cultures were isolated from human resection specimens and from Landrace pigs. Single and cocultures were exposed to porcine and human interleukin (IL)-6 or tumor necrosis factor (TNF)-alpha. Changes in expression of C-reactive protein (CRP), albumin, CCAAT enhancer binding protein (C/EBP)-alpha and C/EBP-beta and metabolic competence of cultured cells was studied by measuring testosterone metabolite production. After human or porcine IL-6 dosing, CRP was induced up to 100-fold in human hepatocyte cultures, while porcine hepatocytes responded marginally (2- to 5-fold). Treatment with human or porcine IL-6 or TNF-alpha resulted in reduced albumin production, albeit at different levels when human and porcine hepatocytes were compared (P = 0.001). Unlike human, porcine hepatocytes produced less of 6alpha-hydroxytestosterone (6alpha-HT) (P < 0.001) and 7alpha-HT (P < 0.001) after human or porcine IL-6 dosing and treatment with species-specific TNF-alpha induced (human hepatocytes) or decreased (porcine hepatocytes) 6beta-HT production (P = 0.021). In coculture with free exchange of metabolites, porcine hepatocytes produced less 6alpha-HT (P = 0.048) and 16alpha-HT (P = 0.033), whereas after treatment with human IL-6 reduced CRP gene and protein expression was observed with human hepatocytes (P = 0.013). In conclusion, species-specific responses of hepatocytes to cytokines and interactions with xenobiotic metabolites may limit the clinical effectiveness of porcine hepatocytes in BAL. PMID- 17133584 TI - Delivery of antioxidative enzyme genes protects against ischemia/reperfusion induced liver injury in mice. AB - Hepatic ischemia/reperfusion (I/R) injury is characterized by the generation of reactive oxygen species (ROS), such as superoxide anions and hydrogen peroxide. The aim of this study is to investigate whether antioxidative gene delivery by our polylipid nanoparticles (PLNP) is an effective approach for prevention of the injury. Polyplexes of extracellular superoxide dismutase (EC-SOD) and/or catalase genes were injected via the portal vein 1 day prior to a warm I/R procedure in mice. The effects of the gene delivery were determined 6 hours after starting reperfusion. PLNP-mediated antioxidative gene delivery led to a marked increase in human EC-SOD and catalase gene expression in the liver. Liver superoxide dismutase (SOD) and catalase activity both increased approximately 10-fold. Increased liver superoxide anion levels caused by the I/R procedure were reduced to normal levels by EC-SOD gene delivery. The overexpression of these 2 antioxidative genes significantly suppressed the I/R-induced elevation of serum alanine aminotransferase (ALT) levels, decreased liver malondialdehyde content, restored glutathione reserve, and improved liver histology. In conclusion, EC-SOD or catalase gene delivery by PLNP resulted in high levels of the transgene activity in the liver, and markedly attenuated hepatic I/R injury. The protection is directly associated with elevated antioxidative enzyme activity as the result of the gene delivery. This novel approach may become a potential therapy to improve graft function and survival after liver transplantation. PMID- 17133585 TI - Clinical and histological efficacy of pegylated interferon and ribavirin therapy of recurrent hepatitis C after liver transplantation. AB - Treatment of recurrent hepatitis C in liver transplant is controversial. The aim of our study was to evaluate the clinical and histological efficacy of pegylated interferon alpha 2b (PEG-IFN) and ribavirin therapy of recurrent hepatitis C after liver transplantation (LT). We prospectively included 47 liver transplant patients with: 1) a positive test for hepatitis C virus (HCV)-ribonucleic acid (RNA) in serum; 2) alanine aminotransferase (ALT) >45 UI/mL; and 3) a liver biopsy showing chronic hepatitis without rejection in the previous 2 months. Patients received PEG-IFN (1.5 microg/kg/week) and ribavirin (800-1,000 mg/day) for 12 months. Follow-up was based on biochemical (ALT), virological (RNA-HCV), and histological (liver biopsy) examinations. Follow-up lasted a minimum of 6 months after the end of antiviral therapy. Sustained virological response (SVR) was achieved in 23% of the patients. A total of 33 (70%) patients had normalized ALT levels at the end of therapy. Inflammatory portal and lobular score declined significantly in patients with SVR (P < 0.05) but not in nonresponder patients. Fibrosis did not change significantly in either group. SVR was significantly associated with low gamma-glutamyltransferase GGT (P = 0.04) and HCV-RNA levels (P = 0.03), a virological response at 12 weeks (P = 0.002) and patient's compliance (P = 0.04). Ten (21%) patients were withdrawn prematurely due to adverse effects. In conclusion, Therapy with PEG-IFN and ribavirin achieved SVR and a significant histological improvement in 23% of liver transplant recipients with chronic hepatitis C. Toxicity is an important drawback of this therapy. PMID- 17133586 TI - The incidence of nonadherence after organ transplant: ensuring that our efforts at counting really count. PMID- 17133588 TI - Liver sinusoidal endothelial cells: their unique role in immune tolerance. PMID- 17133587 TI - In vivo expansion of two distinct dendritic cells in mouse livers and its impact on liver immune regulation. AB - Liver transplant tolerance in pigs, rats, and mice has been disclosed for decades, but the underlying mechanisms are not completely understood. Accumulating data indicate that residing dendritic cells (DC) are important in determining direction of immune responses in the liver. However, our knowledge remains very limited due to the difficulties in obtaining sufficient liver DC. Most of the previous studies were dependent on DC propagated in vitro with growth factors and cytokines. In this study, we adopted an approach to transfect genes into the mouse liver by tail vein injection of plasmid DNA. Transfection with plasmid granulocyte-macrophage colony-stimulating factor markedly expanded liver CD11c(+) DC mainly located in portal regions, while liver B220(+) DC were dramatically generated after injection with plasmid interleukin (IL)-3/CD40L largely present in the lobules. Although both were phenotypically mature and strong T-cell stimulators, CD11c(+)DC induced potent T-cell response while B220(+)DC induced T-cell hyporesponsiveness. Administration of CD11c(+)DC accelerated cardiac allograft rejection, while B220(+)DC significantly prolonged graft survival. This hyporesponsiveness is not due to inhibition of DC/T-cell interaction, but rather through an active process of stimulating T-cell apoptosis. Compared to B220(+) DC that expressed messenger RNA of (TLR) 1, 2, 6, 7, and 9, CD11c(+)DC expressed all TLR 1 to 9. TLR 9 ligation stimulated very high IL-12 in CD11c(+) DC, but high IL-10 and no IL-12 in B220(+) DC. In conclusion, through these mechanisms, liver DC may be actively involved in immune regulation in the liver. PMID- 17133590 TI - The potential role for iron overload and fungal infection in liver transplantation. PMID- 17133591 TI - Neoadjuvant therapies for hepatocellular carcinoma before liver transplantation: a critical appraisal. PMID- 17133592 TI - Knee joint loading differs in individuals with mild compared with moderate medial knee osteoarthritis. AB - OBJECTIVE: To compare the knee joint loading patterns in individuals with differing radiographic grades of knee osteoarthritis (OA) for characterization of the mechanical implications of different structural states, and to compare the knee adduction angular impulse, a measure of gait complementary to the commonly used peak knee adduction moment. METHODS: Asymptomatic subjects (those without knee OA) having a Kellgren/Lawrence (K/L) radiographic severity grade of 0 or 1 (n = 28) and subjects with symptomatic knee OA having K/L grades of 2 (n = 66) or 3 (n = 23) were recruited. Gait analysis was used to calculate the peak external knee adduction moment and the external knee adduction angular impulse for the whole stance and for the 4 subdivisions of stance. RESULTS: Both the peak knee adduction moment and the knee adduction angular impulse increased with K/L radiographic grade (P < 0.05). However, only the knee adduction angular impulse differed between subjects with moderate (grade 3) and those with mild (grade 2) radiographic knee OA (P < 0.05). CONCLUSION: The differences between mild and moderate symptomatic radiographic knee OA are not only structural but also functional, based on the magnitude of load in the medial knee joint. Moreover, knee adduction angular impulse provides additional information beyond that available from the peak knee adduction moment, and may therefore be an important gait parameter to include in OA research. These findings are important for our understanding of the pathophysiologic mechanisms of OA. PMID- 17133593 TI - Effects of chronic growth hormone and insulin-like growth factor 1 deficiency on osteoarthritis severity in rat knee joints. AB - OBJECTIVE: To determine the effects of chronic deficiency of growth hormone (GH) and insulin-like growth factor 1 (IGF-1) on osteoarthritis (OA) severity. METHODS: Thirty-five rats were divided into 4 treatment groups at 4 weeks of age: 1 control group (normal GH/IGF-1 levels [heterozygous]) and 3 groups of dwarves with a genetic mutation that results in GH deficiency. The first dwarf group received GH for 64 weeks (GH replete) and the second received GH until 14 weeks of age, followed by saline for 50 weeks (adult-onset GH/IGF-1 deficiency [AO GHD]). The third dwarf group received saline injections only (lifetime GH deficient [GHD]). Sections of the medial knee joint compartment were graded and measured histologically; data were summarized using factor analysis, and treatment effects were assessed using analysis of variance and adjusting for body weight. RESULTS: Terminal IGF-1 levels and body weights were significantly affected by treatment (P = 0.002 and P < 0.001, respectively). Factor analysis yielded a total of 5 factors, the first 3 of which were not significantly affected by treatment. Factor 4 (weighted by medial tibial plateau articular cartilage width and area) was significantly affected by treatment (P < 0.012), with larger values in the AO-GHD group than in the GHD group (P < 0.05). Factor 5 (weighted primarily by articular cartilage structure and loss of toluidine blue staining scores) also was significantly affected by treatment (P < 0.001), and was significantly lower (less severe lesions) in the GH replete group than in all other treatment groups (P < 0.05). Despite the presence of cartilage lesions, osteophytes and subchondral sclerosis were not observed in GH/IGF-1-deficient animals. CONCLUSION: These results indicate that chronic GH/IGF-1 deficiency causes an increased severity of articular cartilage lesions of OA without the bony lesions normally seen in this disease. PMID- 17133594 TI - Fcgamma receptors directly mediate cartilage, but not bone, destruction in murine antigen-induced arthritis: uncoupling of cartilage damage from bone erosion and joint inflammation. AB - OBJECTIVE: To determine the relationship between synovial inflammation and the concomitant occurrence of cartilage and bone erosion during conditions of variable inflammation using various Fcgamma receptor knockout (FcgammaR(-/-)) mice. METHODS: Antigen-induced arthritis (AIA) was introduced in the knee joints of various FcgammaR(-/-) mice and wild-type controls. Joint inflammation and cartilage and bone destruction levels were determined by histologic analysis. Cathepsin K, RANKL, and osteoprotegerin (OPG) levels were detected by immunolocalization. RESULTS: In FcgammaRIIb(-/-) mice, which lack the inhibiting Fcgamma receptor IIb, levels of joint inflammation and cartilage and bone destruction were significantly higher (infiltrate 93%, exudate 200%, cartilage 100%, bone 156%). AIA in mice lacking activating FcgammaR types I, III, and IV, but not FcgammaRIIb (FcR gamma-chain(-/-) mice), prevented cartilage destruction completely. In contrast, levels of bone erosion and joint inflammation were comparable with their wild-type controls. Of great interest, in arthritic mice lacking activating FcgammaR types I, II, and III, but not IV (FcgammaRI/II/III(-/ ) mice), levels of joint inflammation were highly elevated (infiltrate and exudate, 100% and 188%, respectively). Cartilage destruction levels were decreased by 92%, whereas bone erosion was increased by 200%. Cathepsin K, a crucial mediator of osteoclasts, showed a strong correlation with the amount of inflammation but not with the amount of activating FcgammaR, which was low in osteoclasts. RANKL, but not OPG, levels were higher in the inflammatory cells of arthritic knee joints of FcgammaRI/II/III(-/-) mice versus wild-type mice. CONCLUSION: Activating FcgammaR are crucial in mediating cartilage destruction independently of joint inflammation. In contrast, FcgammaR are not directly involved in bone erosion. Indirectly, FcgammaR drive bone destruction by regulating joint inflammation. PMID- 17133595 TI - Jun activation domain-binding protein 1 binds Smad5 and inhibits bone morphogenetic protein signaling. AB - OBJECTIVE: Bone morphogenetic proteins (BMPs) play an important role in the development and the homeostasis and pathology of cartilage tissue, particularly in the differentiation and anabolic activity of chondrocytes. The present study was undertaken to identify binding partners of the Smad proteins, the intracellular mediators of BMP activity, which might actively modify BMP signaling in chondrocytes. METHODS: Yeast 2-hybrid technology was used to screen a complementary DNA library, constructed from human adult articular cartilage, for molecular binding partners of Smad5, a major intracellular mediator of BMP signaling. Primary interaction partners were verified by coimmunoprecipitation, and the relevance of the interactions to BMP signaling was evaluated by transcriptional reporter assay. Additionally, messenger RNA expression analysis (conventional and quantitative polymerase chain reaction) and immunostaining were performed in adult normal and osteoarthritic articular cartilage. RESULTS: We identified a novel Smad5 interactor, Jun activation domain-binding protein 1 (Jab1), expressed in adult cartilage. The interaction was confirmed in coimmunoprecipitation experiments. Overexpression of Jab1 resulted in an attenuation of BMP-dependent transcriptional responses, suggesting that Jab1 acts as an inhibitor of BMP signaling. CONCLUSION: Jab1 is a newly identified intracellular (negative) modulator of BMP signaling in chondrocytes and other cells. Jab1 represents an interesting molecule for understanding anabolic signaling in chondrocytes, as well as a potential therapeutic target for anabolic activation. Most interestingly, Jab1 appears to crosslink the BMP and interleukin 1 pathways. PMID- 17133596 TI - SPARC, an upstream regulator of connective tissue growth factor in response to transforming growth factor beta stimulation. AB - OBJECTIVE: To differentiate the effects of inhibition of specific small interfering RNA (siRNA) of SPARC (secreted protein, acidic and rich in cysteine) and siRNA of connective tissue growth factor (CTGF) in cultured human fibroblasts, and to identify potential interrelationships between SPARC and CTGF. METHODS: Fibroblasts from skin biopsy specimens of 2 normal individuals were transfected with siRNA of SPARC and siRNA of CTGF. The fibroblasts were stimulated with or without transforming growth factor beta1 (TGFbeta1) and examined by real-time quantitative reverse transcription-polymerase chain reaction to determine the transcription levels of several extracellular matrix genes. RESULTS: After exogenous TGFbeta1 stimulation, both SPARC siRNA and CTGF siRNA showed a protective role against overexpression of collagen genes. Following TGFbeta1 stimulation, SPARC siRNA-transfected fibroblasts showed a greater reduction in expression of the collagen genes compared with CTGF siRNA transfected fibroblasts, as well as a significantly decreased expression of CTGF (P < 0.05). Using linear structure equations to quantitatively model a genetic network based on expression levels of each gene, a positive regulatory role of SPARC on CTGF, COL1A2, COL3A1, COL11A1, and TIMP3 was observed. However, the regulatory role of CTGF on SPARC appeared to be negative and very small, while the positive regulatory effects of CTGF on COL1A2, COL3A1, COL11A1, and TIMP3 were less than those of SPARC. CONCLUSION: The results of this quantitative comparison support the hypothesis that in these cultured fibroblasts, the regulatory effects of SPARC on some major extracellular matrix structural components are greater than those of CTGF. In addition, SPARC appears to regulate CTGF in a predominantly positive manner, while CTGF may act as a negative feedback control on SPARC following TGFbeta stimulation. PMID- 17133597 TI - Steroid hormones strongly support bovine articular cartilage integration in the absence of interleukin-1beta. AB - OBJECTIVE: Posttraumatic integration of articular cartilage at fracture sites is essential for mechanical stability of cartilage, and ruptured cartilage is a prerequisite for early osteoarthritis. This study was undertaken to investigate effects on articular cartilage integration mediated by steroid hormones, interleukin-1beta (IL-1beta), and combinations thereof. METHODS: Articular cartilage blocks were cultured in partial apposition for 2 weeks with ascorbic acid, testosterone, 17beta-estradiol, and dehydroepiandrosterone (DHEA), with or without IL-1beta. Mechanical integration was measured as adhesive strength, i.e., the maximum force at rupture of integrated cartilage blocks divided by the overlap area. Glycosaminoglycan content was used to study synthesized extracellular matrix. RESULTS: Culture in medium without supplements did not lead to integration (adhesive strength 0 kPa). With administration of ascorbic acid (100 microg/ml), the median adhesive strength was 49 kPa. In comparison with ascorbic acid alone, all steroid hormones induced a strong, concentration dependent stimulation of integration (with maximum values observed with DHEA at 3 x 10(-5)M, testosterone at 10(-8)M, and 17beta-estradiol at 10(-11)M). For testosterone and 17beta-estradiol, this was also reflected by an increase of glycosaminoglycan content. Adhesive strength was increased with IL-1beta at 10 pg/ml, but not at 1 pg/ml or 100 pg/ml. In the presence of both IL-1beta and sex hormones, integration of articular cartilage was reduced. CONCLUSION: This is the first study to demonstrate that steroid hormones such as 17beta-estradiol, DHEA, and testosterone stimulate articular cartilage integration. This effect is abrogated by low concentrations of IL-1beta. In the absence of IL-1beta or after neutralization of IL-1beta, steroid hormones might be favorable adjuvant compounds to optimize cartilage integration. PMID- 17133598 TI - Soluble receptor for advanced glycation end products triggers a proinflammatory cytokine cascade via beta2 integrin Mac-1. AB - OBJECTIVE: Receptor for advanced glycation end products (RAGE) is a cell surface molecule that binds a variety of ligands, including high mobility group box chromosomal protein 1 (HMGB-1), a potent proinflammatory cytokine. RAGE-ligand interaction leads to an inflammatory response. A truncated form of the receptor, soluble RAGE (sRAGE), has been suggested to function as a decoy abrogating cellular activation, but its endogenous activity is not fully understood. We undertook this study to assess the properties of sRAGE in vivo and in vitro and to analyze the role of sRAGE in HMGB-1-induced arthritis. METHODS: Mice were injected intraarticularly with HMGB-1 and treated systemically with sRAGE prior to histologic joint evaluation. All animals were subjected to peritoneal lavage to assess the local effect of sRAGE treatment. For in vitro studies, mouse splenocytes were incubated with sRAGE followed by assessment of NF-kappaB activation and cytokine production. The chemotactic properties of sRAGE were investigated using in vitro migration assay. RESULTS: Soluble RAGE was determined to have proinflammatory properties since it gave rise to production of interleukin-6, tumor necrosis factor alpha, and macrophage inflammatory protein 2. This effect was triggered by interaction with leukocyte beta2 integrin Mac-1 and was mediated via NF-kappaB. Systemic treatment with sRAGE significantly down regulated HMGB-1-triggered arthritis, but the observed effect was due to a deviation of the inflammatory response from the joint to the peritoneal cavity rather than a genuine antiinflammatory effect. Apart from its proinflammatory properties, sRAGE was proven to act as a chemotactic stimulus for neutrophils. CONCLUSION: We conclude that sRAGE interacts with Mac-1, thereby acting as an important proinflammatory and chemotactic molecule. PMID- 17133599 TI - Relationship of antiphospholipid antibodies to cardiovascular manifestations of systemic lupus erythematosus. AB - OBJECTIVE: Although antiphospholipid antibodies (aPL) are associated with arterial and venous thrombosis in systemic lupus erythematosus (SLE), the extent to which they influence other cardiovascular manifestations is either controversial or uncertain. We undertook this study to examine the relationships of aPL with valvular, myocardial, and arterial disease in SLE. METHODS: Two hundred patients in an SLE registry, recruited at the time of outpatient visits, underwent comprehensive interviews, physical examinations, laboratory assessments, echocardiography to assess left ventricular (LV) and valvular status, carotid ultrasonography to detect atherosclerosis (discrete plaque), and radial applanation tonometry to measure arterial stiffness. RESULTS: Antiphospholipid antibodies were present(defined as IgG or IgM anticardiolipin > or =40 IU/ml or the presence of lupus anticoagulant) in 42 patients (21%). Mitral valve nodules and moderate-to-severe mitral regurgitation were more common in aPL positive patients (both 14.3% versus 4.4%; P = 0.02). Thirty-one percent of patients with high titers of IgG aPL (>80 IU/ml) had mitral valve nodules, compared with 20% of patients with mildly to moderately elevated levels of IgG aPL (16-80 IU/ml) and 4% of patients without IgG aPL (overall P < 0.001). Levels of soluble tumor necrosis factor receptors were higher in the presence of both aPL and mitral valve nodules. LV dimensions, systolic function, and carotid artery stiffness as well as prevalences of Raynaud's phenomenon, pulmonary hypertension, and atherosclerosis were similar in aPL-positive and aPL-negative patients. CONCLUSION: Antiphospholipid antibodies in SLE are associated with mitral valve nodules and significant mitral regurgitation, possibly due to valvular endothelial cell activation. However, in this population, they are not associated with evidence of myocardial hypertrophy, systolic dysfunction, coronary or carotid atherosclerosis, or other vascular abnormalities. PMID- 17133600 TI - Association of a transmembrane polymorphism of Fcgamma receptor IIb (FCGR2B) with systemic lupus erythematosus in Taiwanese patients. AB - OBJECTIVE: To investigate the possible association of the Fcgamma receptor IIb (FcgammaRIIb) Ile/Thr187 transmembrane domain polymorphism, which significantly affects receptor signaling, with susceptibility to systemic lupus erythematosus (SLE) in Taiwanese patients. METHODS: We used matrix-assisted laser desorption ionization-time-of-flight mass spectrometry to genotype 351 Taiwanese SLE patients and 372 age- and sex-matched healthy individuals from the same geographic area. Allele frequencies and genotype distributions were compared between the patients and controls, both as an aggregate and as stratified by sex, autoantibody profile, and clinical parameters. A combined analysis was conducted to assess the FCGR2B Thr187 allele as a common risk factor in different ethnic populations. RESULTS: The minor Thr187 allele was significantly associated with SLE in Taiwanese subjects (P = 0.017, odds ratio [OR] 1.989 [95% confidence interval (95% CI) 1.119-3.553]). Interestingly, male SLE patients showed enrichment of the Thr/Thr187 genotype (24%; 7 of 29) as compared with female SLE patients (10%; 32 of 322) (P = 0.043, OR 2.884 [95% CI 1.028-7.839]). Additionally, SLE patients with Thr/Thr187 and Ile/Thr187 genotypes were more likely to have pleural effusions (P = 0.038, OR 1.874 [95% CI 1.033-3.411]) and anti-SSA/Ro antibody production (P = 0.046, OR 2.221 [95% CI 1.013-4.897]). Combined analysis of 4 groups of Asian patients strongly supported the association of the FCGR2B Thr187 allele with the lupus phenotype (P = 0.000159). CONCLUSION: The FcgammaRIIb transmembrane polymorphism is a strong disease susceptibility candidate in epistasis with other genetic effects in Taiwanese and other Asian populations. It may also play a more prominent role in male patients with SLE. PMID- 17133601 TI - Comparison of global gene expression between porcine testis tissue xenografts and porcine testis in situ. AB - Testis tissue from immature mammalian donor animals, grafted ectopically to immunodeficient mouse hosts, can undergo complete spermatogenesis with the production of fertilization-competent spermatozoa. To further characterize testis tissue xenografts as a model for testis function in situ, the objective of this study was to compare gene expression between porcine testis tissue xenografts and testis tissue in situ. Pieces of testis tissue from 1-week-old piglets were grafted onto immunodeficient male mice and a littermate piglet was raised for comparison as control. Complete spermatogenesis was present in the testis tissue xenografts at 8 months after transplantation into mouse hosts and in the 8-month old control porcine testis tissue. Total RNA was isolated from xenografts and control tissue, and the RNA was labeled and hybridized to the porcine genome array. By analyzing the expression of 23,256 transcripts, we found that 71 genes were differentially expressed with at least a fourfold difference between xenografts and control tissue. Interestingly, none of the 56 transcripts present on the array that were annotated in porcine testis showed differential expression between xenografts and control testis. This analysis indicates that global gene expression in porcine testis xenografts appears comparable to testis tissue in situ. These findings support the hypothesis that testis tissue xenografts can provide a representative model to study mammalian spermatogenesis. PMID- 17133602 TI - Using the histone H2a transcript as an endogenous standard to study relative transcript abundance during bovine early development. AB - The requirement for sample standardization is basic to any relative RNA abundance assessment. In 2002, we published on the RNA abundance profiling of several housekeeping targets during early bovine embryonic development. It was then concluded that histone H2a was the most stable transcript across the studied developmental period. Since that time, several teams have applied this information, yet neglected to use the published set of primers. Here, we show that these other primer sets do not target the same histone H2a variant. Within the present report, the RNA abundance profiles of their respective targets, for example, histone H2a.1, H2a.z, and H2a.o were measured in developmental series spanning the immature oocyte to the blastocyst stage embryo. In order to more clearly define the conditions that impact the RNA abundance level measurement of these candidates, the state of polyadenylation and the origin of the transcript either from de novo transcription or from maternal stocks were taken into consideration. The histone H2a.z transcript is principally regulated by de novo transcription following embryonic genome activation, whereas the levels of H2a.1 and H2a.o variants are largely of maternal origin, conferring a more stable profile throughout the studied developmental window. The histone H2a.z was found solely in a polyadenylated state whereas the H2a.1 and H2a.o were found to be more abundant in a nonadenylated form. The presence of serum in the in vitro embryo production system also had some impact on the histone H2a.1 RNA level at the blastocyst stage. PMID- 17133603 TI - Dynamin 2 associates with complexins and is found in the acrosomal region of mammalian sperm. AB - Previous data showed that complexin I, a SNARE regulatory protein, is localized in and/or around the acrosome and is necessary for the acrosome reaction in sperm. To understand how complexin I regulates the acrosome reaction, we used complexin-GST pulldown assays to identify interacting proteins. We showed that both complexins I and II bound mouse sperm dynamin 2. Dynamin 2 is a 100 kDa GTPase essential to many aspects of endocytosis but its potential role in exocytosis is unknown. Dynamin 2 is expressed in rat testis and widely expressed in other tissues; however, the function of dynamin 2 in germ cells is uncertain. Dynamin 2 protein was detected in mouse testis and was most abundant in or around the developing acrosome of spermatids. In addition, dynamin 2 was co-localized with complexin I in the acrosomal region of mammalian sperm. Its co-localization and interaction with complexin I suggest that dynamin 2 may play a role during acrosome formation and/or acrosomal exocytosis. PMID- 17133604 TI - Crypticity and functional distribution of the membrane associated alpha-L fucosidase of human sperm. AB - Two distinctive isoforms of the enzyme alpha-L-fucosidase are found within human semen in substantial amounts, suggesting specialized functions during reproduction. The membrane-associated isozyme of human sperm cells was previously characterized biochemically, and here we report on its subcellular localization. Intact, detergent permeabilized, capacitated, and acrosome-reacted sperm were investigated using antifucosidase immunofluorescence, binding of the fluorescent fucosylated glycoconjugate RITC-BSA-fucose (RBF), and enzyme activity in the presence and absence of selected inhibitors. Both immunolocalization and RBF binding show that fucosidase is broadly distributed over the membrane systems of human sperm, but is relatively enriched within the equatorial segment. Upon detergent treatment or induction of acrosome reaction (AR), a portion of enzyme activity is recoverable in the supernatant, presumably associated with released remnants of the outer acrosomal membrane. Surprisingly, cell-bound enzyme activity increases sharply following permeabilization of intact sperm, representing cryptic fucosidase that is relatively stable and corresponds with strong fluorescence in the equatorial segment and other sperm membranes. These observations support the notion that the fucosidase has a role in the intimate species signature interactions between sperm and oocyte. PMID- 17133605 TI - Ultrastructural analysis reveals striking differences of intercellular contact lengths in bovine embryos produced in vivo, in vitro and by somatic cell nuclear transfer. AB - Cellular coherence and communication, thus cell-to-cell contact is an indispensable premise to sustain the formation of complex, multi-cellular organisms. We have analyzed intercellular contact lengths in NT-cloned bovine embryos compared to the in vivo or in vitro produced counterparts. Therefore, ultrastructural analysis was carried out by transmission electron microscopy (TEM) at the 8-cell and blastocyst stage of development. To obtain embryos generated in vivo, oviducts of superovulated cows were flushed 3 days after insemination, subsequent to slaughter. Standard in vitro maturation (IVM) and fertilization (IVF) were utilized to obtain in vitro embryos. Cloned embryos by somatic nuclear transfer were produced by the handmade cloning (HMC) procedure. The points of apposition/focal contact points (CPs) between the blastomeres were of the shortest order in cloned embryos (236 +/- 135 nm) and of highest order in the in vivo produced embryos (2,085 +/- 1,540 nm), although no significant differences regarding the blastomere sizes in the various groups of 8-cell embryos could be established. In summary, the CP lengths in case of in vitro and in vivo 8-cell embryos were, on an average, five or nine times longer, respectively, than in the case of the cloned embryos. These differences of CP lengths vanished in embryos reaching the blastocyst stage of embryonic development in all the three groups of embryos. The observed differences of intercellular contact length at distinct stages of embryonic development could be responsible for differences in intercellular communication between the blastomeres at the beginning of cellular differentiation. These may be one reason for the lower developmental competence of cloned (NT) embryos. PMID- 17133606 TI - Domain 1 of the urokinase-type plasminogen activator receptor is required for its morphologic and functional, beta2 integrin-mediated connection with actin cytoskeleton in human microvascular endothelial cells: failure of association in systemic sclerosis endothelial cells. AB - OBJECTIVE: In systemic sclerosis (SSc) microvascular endothelial cells (MVECs), angiogenesis is blocked by matrix metalloproteinase 12-dependent cleavage of domain 1 of the urokinase-type plasminogen activator receptor (uPAR). Since integrins are associated with the invasive activity of uPAR in angiogenesis, this study was undertaken to show whether full-size and truncated uPAR are differentially associated with integrins and with motor components of the cytoskeleton. METHODS: SSc and normal MVECs were isolated from human skin biopsy specimens and studied by confocal laser scanning microscopy and immunoprecipitation to assess the mechanisms of association of truncated and full size uPAR with integrins and the actin cytoskeleton. The integrin composition of the MVECs was studied by reverse transcription-polymerasechain reaction. Cell migration and capillary morphogenesis were studied on fibrinogen substrates. Involvement of Rac and Cdc42 was evaluated by Western blotting. RESULTS: Only full-size uPAR showed a connection with the actin cytoskeleton in ECs. This connection was mediated by the uPAR-associated alphaMu- and alphaX-subunits of beta2 integrin, and was absent from SSc MVECs. The cleaved uPAR was not associated with beta2 integrins or with actin. beta3 integrins were associated with both the full-size and cleaved uPAR at focal contacts. The uncoupling of uPAR from beta2 integrins in SSc MVECs impaired the activation of Rac and Cdc42 (thus inhibiting their mediation of uPAR-dependent cytoskeletal rearrangements and cell motility) and blocked the integrin-engagement-delivered signals to the actin cytoskeleton. Invasion and capillary morphogenesis on fibrinogen-coated substrates indicated that ligation of uPAR by uPA empowers the beta2/beta3 integrin-dependent invasion of fibrinogen, and that this system is impaired in SSc MVECs. CONCLUSION: The reduced angiogenic properties of SSc MVECs can be explained by the effects of uPAR truncation and the subsequent loss of the beta2 integrin-mediated connection of uPAR with the actin cytoskeleton in these ECs. PMID- 17133607 TI - A novel autoantigen to differentiate limited cutaneous systemic sclerosis from diffuse cutaneous systemic sclerosis: the interferon-inducible gene IFI16. AB - OBJECTIVE: To investigate the presence and clinical significance of autoantibodies against the interferon-inducible gene IFI16 in systemic sclerosis (SSc), systemic lupus erythematosus (SLE), and other autoimmune diseases. METHODS: Immunohistochemical analysis was used to evaluate the expression of IFI16 in skin biopsy specimens obtained from patients with SSc and patients with SLE. Levels of antibodies against IFI16 in sera from 82 patients with SSc and 100 patients with SLE were determined by enzyme-linked immunosorbent assay. Other autoimmune diseases such as primary Sjogren's syndrome (SS), rheumatoid arthritis (RA), chronic urticaria, and hepatitis C virus (HCV) infection were also examined. RESULTS: Expression of IFI16 was greatly increased and was ubiquitous in all layers of the epidermis and in the dermal inflammatory infiltrates of lesional skin from both patients with SLE and patients with SSc. Patients with SLE, those with primary SS, and those with SSc exhibited significantly higher anti-IFI16 IgG antibody levels compared with normal controls (for SLE, P < 0.002; for primary SS, P < 0.001; for SSc, P < 0.0005). Anti-IFI16 titers above the ninety-fifth percentile for control subjects were observed in 26% of the patients with SLE, 50% of those with primary SS, and 21% of those with SSc (28% of patients with limited cutaneous SSc [lcSSc] versus 4% of patients with diffuse cutaneous SSc [dcSSc]). In contrast, the prevalence of anti-IFI16 was 4% in patients with RA, 5% in those with chronic urticaria, and 13% in those with HCV infection. CONCLUSION: The results of this study provide evidence that an IFN inducible gene, IFI16, may be involved in the pathophysiologic mechanisms of connective tissue disorders such as SSc. Moreover, a strict correlation with lcSSc was also demonstrated, thus providing a novel tool in the differential diagnosis of lcSSc from dcSSc. PMID- 17133608 TI - Association of the PTPN22 R620W polymorphism with anti-topoisomerase I- and anticentromere antibody-positive systemic sclerosis. AB - OBJECTIVE: To determine any associations of the PTPN22 R620W single-nucleotide polymorphism (SNP) with systemic sclerosis (SSc) or with anticentromere antibody (ACA)-positive or anti-topoisomerase I (anti-topo I) antibody-positive SSc, in a case-control study of US white, black, Hispanic, and Choctaw Indian individuals. METHODS: A total of 850 white, 130 black, 120 Hispanic, and 20 Choctaw Indian patients with SSc were compared with 430 white, 164 black, 146 Hispanic, and 76 Choctaw Indian control subjects, respectively. All subjects were living in the US. PTPN22 SNP (rs2476601) genotyping was performed by TaqMan 5' allelic discrimination assay and pyrosequencing. RESULTS: The PTPN22 CT/TT genotype showed significant association with anti-topo I antibody-positive SSc in white patients (odds ratio [OR] 2.21, 95% confidence interval [95% CI] 1.3-3.7) and with ACA-positive white patients with SSc (OR 1.70, 95% CI 1.1-2.7). Frequency of the PTPN22*T allele also showed significant association with anti-topo I antibody positive SSc in white patients (OR 2.03, 95% CI 1.3-3.2). When data for patients in the 3 ethnic groups (black, white, and Hispanic) were combined, a significant association with both genotype and allele frequencies was observed, suggesting a trend toward association in ACA-positive and anti-topo I antibody-positive SSc. Stepwise logistic regression analysis (controlled for the confounding effects of sex and race) showed that the PTPN22 CT/TT genotype was associated with a significantly higher risk of SSc compared with the CC genotype (for patients with SSc, OR 1.64, 95% CI 1.2-2.2; for ACA-positive patients with SSc, OR 1.63, 95% CI 1.0-2.6; for anti-topo I antibody-positive SSc, OR 2.33, 95% CI 1.5-3.7). CONCLUSION: Our results indicate that the PTPN22 R620W polymorphism is associated with ACA-positive and anti-topo I antibody-positive subsets of SSc and represents a risk factor in both white patients and black patients. The association of subsets of SSc with the PTPN22 R620W polymorphism further strengthens the classification of SSc within the spectrum of autoimmune diseases and strongly suggests the involvement of common susceptibility genes and similarly disordered immunoregulatory pathways. PMID- 17133609 TI - Lung transplantation in scleroderma compared with idiopathic pulmonary fibrosis and idiopathic pulmonary arterial hypertension. AB - OBJECTIVE: Lung transplantation is a viable, life-saving intervention for several primary pulmonary disorders complicated by severe lung dysfunction. This study was undertaken to evaluate whether patients with systemic sclerosis (scleroderma), a systemic autoimmune rheumatic disorder, would receive similar benefit from this intervention. METHODS: Survival following lung transplantation was examined at 2 university medical centers among 29 patients with scleroderma as compared with 70 patients with idiopathic pulmonary fibrosis (IPF) and 38 with idiopathic pulmonary arterial hypertension (IPAH), the latter groups representing pathologically related primary pulmonary disorders. The end point was death from any cause. Risk of mortality in patients with scleroderma was compared with that in patients with IPF or IPAH, with adjustment for demographic and clinical parameters. RESULTS: During 2 years of followup, 11 patients with scleroderma (38%), 23 with IPF (33%), and 14 with IPAH (37%) died. Cumulative survival at 6 months posttransplantation was 69% in the scleroderma group compared with 80% in the IPF group (log-rank P = 0.21) and 79% in the IPAH group (P = 0.38). The estimated risk of mortality at 6 months was increased in patients with scleroderma compared with those with IPF (relative risk [RR] 1.70, 95% confidence interval [95% CI] 0.74-3.93) and those with IPAH (RR 1.52, 95% CI 0.59-3.96), but the differences were not statistically significant. Over the following 18 months, there was convergence in the survival rates such that cumulative survival at 2 years was comparable, at approximately 64%, among all 3 groups. CONCLUSION: Patients with scleroderma who are recipients of lung transplantation experience similar rates of survival 2 years after the procedure compared with those with IPF or IPAH. Lung transplantation may represent a viable therapeutic option to consider for patients with end-stage lung disease due to scleroderma. PMID- 17133610 TI - A multicenter, prospective, randomized, double-blind, placebo-controlled trial of corticosteroids and intravenous cyclophosphamide followed by oral azathioprine for the treatment of pulmonary fibrosis in scleroderma. AB - OBJECTIVE: The lack of randomized controlled trials (RCTs) in pulmonary fibrosis in systemic sclerosis (SSc) has hampered an evidence-based approach to treatment. This RCT was undertaken to investigate the effects of intravenous (IV) cyclophosphamide (CYC) followed by azathioprine (AZA) treatment in pulmonary fibrosis in SSc. METHODS: Forty-five patients were randomized to receive low-dose prednisolone and 6 infusions (monthly) of CYC followed by oral AZA, or placebo. Primary outcome measures were change in percent predicted forced vital capacity (FVC) and change in single-breath diffusing capacity for carbon monoxide (DLCO). Secondary outcome measures included changes in appearance on high-resolution computed tomography and dyspnea scores. An intent-to-treat statistical analysis was performed. RESULTS: At baseline, there were no significant group differences in factors linked to outcome, including severity of pulmonary fibrosis and autoantibody status. Sixty-two percent of the patients completed the first year of treatment. Withdrawals included 9 patients (6 from the placebo group) with significant decline in lung function, 2 with treatment side effects (both from the active treatment group), and 6 with non-trial-related comorbidity. No hemorrhagic cystitis or bone marrow suppression was observed. Estimation of the relative treatment effect (active treatment versus placebo) adjusted for baseline FVC and treatment center revealed a favorable outcome for FVC of 4.19%; this between-group difference showed a trend toward statistical significance (P = 0.08). No improvements in DLCO or secondary outcome measures were identified. CONCLUSION: This trial did not demonstrate significant improvement in the primary or secondary end points in the active treatment group versus the group receiving placebo. However, for FVC there was a trend toward statistical significance between the 2 groups. This suggests that treatment of pulmonary fibrosis in SSc with low-dose prednisolone and IV CYC followed by AZA stabilizes lung function in a subset of patients with the disease. Therapy was well tolerated with no increase in serious adverse events. PMID- 17133611 TI - Systemic sclerosis in childhood: clinical and immunologic features of 153 patients in an international database. AB - OBJECTIVE: To determine the clinical and immunologic features of systemic sclerosis (SSc) in a large group of children and describe the clinical evolution of the disease and compare it with the adult form. METHODS: Data on 153 patients with juvenile SSc collected from 55 pediatric rheumatology centers in Europe, Asia, and South and North America were analyzed. Demographic, clinical, and immunologic characteristics of children with juvenile SSc at the onset, at diagnosis, and during the disease course were evaluated. RESULTS: Raynaud's phenomenon was the most frequent symptom, followed by skin induration in approximately 75% of patients. Musculoskeletal symptoms were present in one-third of patients, and the most frequently involved internal organs were respiratory and gastrointestinal, while involvement of renal, cerebral, and cardiovascular systems was extremely rare. Antinuclear antibodies were present in the sera of 81% of patients. Anti-topoisomerase I (Scl-70) and anticentromere antibodies were found to be positive in 34% and 7.1% of patients, respectively. Involvement of the respiratory, gastrointestinal, and cardiovascular systems was more frequent and occurred earlier in patients who died than in those who survived. Compared with the adult form, juvenile SSc appears to be less severe, with the involvement of fewer internal organs, particularly at the time of diagnosis, and has a less characterized immunologic profile. CONCLUSION: This study provides information on the largest collection of patients with juvenile SSc ever reported. Juvenile SSc appears to be less severe than in adults because children have less internal organ involvement, a less specific autoantibody profile, and a better long-term outcome. PMID- 17133612 TI - Immunogenetic risk and protective factors for juvenile dermatomyositis in Caucasians. AB - OBJECTIVE: To define the relative importance (RI) of class II major histocompatibility complex (MHC) alleles and peptide binding motifs as risk or protective factors for juvenile dermatomyositis (DM), and to compare these with HLA associations in adult DM. METHODS: DRB1 and DQA1 typing was performed in 142 Caucasian patients with juvenile DM, and the results were compared with HLA typing data from 193 patients with adult DM and 797 race-matched controls. Random Forests classification and multiple logistic regression were used to assess the RI of the HLA associations. RESULTS: The HLA-DRB1*0301 allele was a primary risk factor (odds ratio [OR] 3.9), while DQA1*0301 (OR 2.8), DQA1*0501 (OR 2.1), and homozygosity for DQA1*0501 (OR 3.2) were additional risk factors for juvenile DM. These risk factors were not present in patients with adult DM without defined autoantibodies. DQA1 alleles *0201 (OR 0.37), *0101 (OR 0.38), and *0102 (OR 0.51) were identified as novel protective factors for juvenile DM, the latter 2 also being protective factors in adult DM. The peptide binding motif DRB1 (9)EYSTS(13) was a risk factor, and DQA1 motifs F(25), S(26), and (45)(V/A)W(R/K)(47) were protective. Random Forests classification analysis revealed that among the identified risk factors for juvenile DM, DRB1*0301 had a higher RI (100%) than DQA1*0301 (RI 57%), DQA1*0501 (RI 42%), or the peptide binding motifs. In a logistic regression model, DRB1*0301 and DQA1*0201 were the strongest risk and protective factors, respectively, for juvenile DM. CONCLUSION: DRB1*0301 is ranked higher in RI than DQA1*0501 as a risk factor for juvenile DM. DQA1*0301 is a newly identified HLA risk factor for juvenile DM, while 3 of the DQA1 alleles studied are newly identified protective factors for juvenile DM. PMID- 17133613 TI - Engineered blood and lymphatic capillaries in 3-D VEGF-fibrin-collagen matrices with interstitial flow. AB - In vitro endothelial cell organization into capillaries is a long standing challenge of tissue engineering. We recently showed the utility of low level interstitial flow in guiding the organization of endothelial cells through a 3-D fibrin matrix-containing covalently bound vascular endothelial growth factor (VEGF). Here this synergistic phenomenon was extended to explore the effects of matrix composition on in vitro capillary morphogenesis of human blood versus lymphatic endothelial cells (BECs and LECs). Different mixtures of fibrin and collagen were used in conjunction with constant concentrations of matrix-bound VEGF and slow interstitial flow over 10 days. Interestingly, the BECs and LECs each showed a distinct preference in terms of organization for matrix composition: LECs organized the most extensively in a fibrin-only matrix, while BEC organization was optimized in the compliant collagen-containing matrices. Furthermore, the BECs and LECs produced architecturally different structures; while BECs organized in thick, branched networks containing wide lumen, the LECs were elongated into slender, overlapping networks with fine lumen. These data demonstrate the importance of the 3-D matrix composition in facilitating and coordinating BEC and LEC capillary morphogenesis, which is important for in vitro vascularization of engineered tissues. PMID- 17133616 TI - Survey of medication adherence in patients with schizophrenia--Korean ADHES data. AB - BACKGROUND: This study evaluated the partial adherence to medication in schizophrenic patients in Korea. METHODS: The Adherence in Schizophrenia (ADHES) survey was conducted worldwide, including Asian countries. Through questionnaires for clinicians, caregivers and patients, information about medication adherence in patients with schizophrenia and factors affecting adherence were identified. This survey involved 131 psychiatrists from 80 psychiatry clinics and 2824 patients with schizophrenia were enrolled. RESULTS: Partial adherence in patients with schizophrenia was prevalent (over 60%) in Korea. From the psychiatrists' point of view, the most prevalent factors associated with partial adherence in their patients were poor awareness of the illness (85%) and embarrassment at having to take medication daily (80%). Psychiatrists believed that most patients (83%) needed help from someone to remind them to take the medication regularly. Of patients, 57% reported feeling upset at having to take medication daily and 76% of caregivers reported preferring long-acting medications. CONCLUSION: Based on the study results, a specific strategy to deal successfully with the prevalent partial adherence to medication in patients with schizophrenia should be developed, and long-acting medication may be one solution to improve partial adherence problems. PMID- 17133618 TI - Ageing in yeast does not enhance prion generation. AB - The yeast prions [URE3] and [PSI(+)] are self-propagating amyloids of Ure2p and Sup35p, respectively. The analogous transmissible spongiform encephalopathies of mammals and other amyloidoses are largely diseases of later life. From normal strains lacking the prions, we isolated old cells and measured the frequency of de novo [URE3] and [PSI(+)] prion generation. We find no evidence that ageing of yeast increases the frequency of prion occurrence. PMID- 17133619 TI - Pooling batches in drug stability study by using constant-width simultaneous confidence bands. AB - One important study objective in drug stability studies is to estimate the shelf life of a drug. A key statistical problem involved in this is how to assess the practical equivalence of different batches of the same drug so that different batches can be subgrouped to produce a single shelf-life for the drug. In this paper constant-width simultaneous confidence bands are proposed to quantify the magnitude of difference between different batches, with a particular view to establish the practical equivalence of different batches. This approach is suitable for the situation that the intercepts and slopes of the regression lines for the batches cannot be assumed to be equal. It is shown how constant-width simultaneous confidence bands can be easily constructed for the multiple comparison of several general linear regression models. In particular, it is shown that constant-width simultaneous confidence bands have a better chance to establish the equivalence than, and so are preferable to, the hyperbola-shaped simultaneous confidence bands considered. PMID- 17133620 TI - Substrate-inducible versions of internal alternative NADH: ubiquinone oxidoreductase from Yarrowia lipolytica. AB - In standard laboratory strains of the obligate aerobic yeast Yarrowia lipolytica, respiratory chain complex I (proton-translocating NADH : ubiquinone oxidoreductase) is an essential enzyme, since alternative NADH dehydrogenase activity is located exclusively at the external face of the mitochondrial inner membrane. Deletions and other loss-of-function mutations in genes for nuclear coded subunits of complex I can be obtained only when an internal version of the latter enzyme, termed NDH2i, is introduced. In contrast to recent findings with Neurospora crassa, external alternative NADH dehydrogenase activity is dispensable in complex I deletion strains of Y. lipolytica. We used regulable promoters to create strains which express internal alternative NADH dehydrogenase in a substrate-dependent manner. The ability to switch between complex I dependent and -independent mode of growth simply by changing the carbon source is an important prerequisite for screens for both loss-of-function and inhibitor resistance mutation. The isocitrate lyase promoter (pICL1), in combination with a NDH2i allele that results in reduced expression and activity, was most promising. In the presence of complex I inhibitors, this construct allowed growth on acetate, but not on glucose minimal media. A somewhat higher background was observed with the acyl-CoA oxidase 2 (pPOX2) promoter on glucose minimal media. PMID- 17133621 TI - Oxygen and carbon source-regulated expression of PDC and ADH genes in the respiratory yeast Pichia anomala. AB - We amplified, sequenced and studied the transcriptional regulation of genes of the alcoholic fermentation pathway in the biocontrol and non-Saccharomyces wine yeast, Pichia anomala. Two ADH isogenes, PaADH1 and PaADH2, and one PDC gene, PaPDC1, were amplified from genomic P. anomala DNA by a two-step PCR approach, using degenerated primers against conserved regions of the respective genes for cloning core regions, and PCR-based gene walking for cloning the respective 5' and 3'-ends. According to sequence analysis, ADH1 and PDC1 are most likely cytoplasmatic proteins, while ADH2 is most probably localized in the mitochondria. PaADH1 was expressed during aerobic growth on glucose, ethanol and succinate, but was nine-fold upregulated in response to oxygen limitation when grown on glucose. The gene seems to be involved in both production and consumption of ethanol. Only low expression of PaADH2 was detected during growth on glucose and ethanol, but it was highly expressed during growth on the non fermentable carbon source succinate and repressed by the addition of glucose. PaPDC1 was expressed during aerobic growth on glucose and was upregulated four fold in response to oxygen limitation. PaPDC1 expression was lower in cells grown on ethanol and succinate than on glucose and was up- regulated two- and four fold, respectively, after glucose addition. Our results demonstrate that transcription of genes of the fermentative pathway is regulated by hypoxia and carbon source but posttranscriptional regulation may play a major role in regulating the metabolic flux. PMID- 17133622 TI - Evaluation of an on-target sample preparation system for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in conjunction with normal flow peptide high-performance liquid chromatography for peptide mass fingerprint analyses. AB - Large-scale mass spectrometry (MS)-based proteomic analyses require high throughput sample preparation techniques due to the increasing numbers of samples that make up a typical proteomics experiment. Moreover, extensive sample pre treatment steps are necessary prior to MS acquisition for even the most rapid and robust MS-based proteomics methodology, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS followed by peptide mass fingerprinting (PMF) analysis. These include sample purification and fractionation, removal of digestion buffers or solvents, and spotting of sample with matrix onto the MALDI target. These multiple steps of time-consuming sample handling can result in high overall analysis costs and the likelihood of sample contamination and loss. In order to overcome some of these limitations in sample processing, we have investigated the use of a novel, simple, inexpensive 96-well elastomeric array that affixes to a MALDI target to create an on-target 96-well plate that accommodates a high solution volume (ca. 200 microL), thereby enabling the on-target processing of samples for MALDI-TOFMS. We explored several factors that influence MALDI sample preparation: type of matrix, solution volume, solution organic composition, solution drying rates and matrix/analyte co crystallization methods. We also investigated the use of the 96-well elastomeric device for coupling MALDI-TOFMS analysis directly to high flow rate (1 mL/min) reversed-phase (rp)-HPLC. By developing an optimized, robust sample preparation protocol, we were able to obtain mass spectra with a high signal-to-noise ratio from peptide standards present at the 50-fmol level in large starting volumes of solution. PMF analyses were possible from 1-pmol and 500-fmol protein-digest standards. Coupling the device to high-flow HPLC (750 microL/min) yielded a robust and semi-automated means to obtain enhanced MALDI-TOFMS data at 500 ng of protein digest. These methodologies developed for this simple, on-target, elastomeric device show promise for streamlining the sample preparation process from HPLC to MALDI-MS. PMID- 17133623 TI - In vitro characterization of the Mig1 repressor from Saccharomyces cerevisiae reveals evidence for monomeric and higher molecular weight forms. AB - The Mig1 DNA-binding protein of Saccharomyces cerevisiae was expressed and purified from yeast and the physical properties were characterized by several methods, including gel filtration, sucrose gradient sedimentation and native gel electrophoresis. Purified Mig1 exists as a monomer with a Stokes' radius of 48 A and a sedimentation coefficient of 3.55 S. Mig1 has an elongated shape with a frictional coefficient of 1.83. The K(d) of purified Mig1 for the SUC2 A site is 2.8 nM and for SUC2 B site 25.8 nM; these values were similar for Mig1 purified from repressed and derepressed cells. Full-length Mig1 expressed in yeast binds more tightly to SUC2 B than bacterially expressed GST-Mig1. Sucrose gradient sedimentation resolved a larger molecular weight form of Mig1 in whole-cell extracts that was not seen in purified samples and may represent a complex with another protein. This complex is found within the nucleus and is seen only in repressed cells. Mig1 exists in multiple phosphorylation states and only less phosphorylated forms of Mig1 are associated with this complex. PMID- 17133625 TI - Current awareness on yeast. PMID- 17133624 TI - Arabidopsis thaliana CHX17 gene complements the kha1 deletion phenotypes in Saccharomyces cerevisiae. AB - AtChx17p is a putative K(+)/H(+) exchanger from Arabidopsis thaliana, expressed in the roots and probably involved in K(+) acquisition and homeostasis. AtCHX17 cDNA complements the phenotypes of the kha1Delta mutation in S. cerevisiae cells: a growth defect at increased pH and hygromycin sensitivity. The localization of GFP-tagged AtChx17 protein in yeast cells is similar to that of ScKha1p: a bold dotted pattern inside the cells resembling the Golgi fluorescence markers. These results show that (a) the proteins AtChx17 and ScKha1 could have similar functions and (b) S. cerevisiae kha1 deletion mutants could serve for the heterologous expression and characterization of plant transporters. The results of this work are evidence that a S. cerevisiae strain with deletions of genes encoding alkali-metal-cation/H(+) antiporters (i.e. Nha1p, Nhx1p, Kha1p) could be an ideal tool for expression and functional analysis of any type of similar plant antiporters (plasma membrane, endosomal/prevacuolar and Golgi). PMID- 17133626 TI - Bayesian estimation of false-negative rate in a clinical trial of sentinel node biopsy. AB - Estimating the false-negative rate is a major issue in evaluating sentinel node biopsy (SNB) for staging cancer. In a large multicentre trial of SNB for intra operative staging of clinically node-negative breast cancer, two sources of information on the false-negative rate are available.Direct information is available from a preliminary validation phase: all patients underwent SNB followed by axillary nodal clearance or sampling. Of 803 patients with successful sentinel node localization, 19 (2.4 per cent) were classed as false negatives. Indirect information is also available from the randomized phase. Ninety-seven (25.4 per cent) of 382 control patients undergoing axillary clearance had positive axillae. In the experimental group, 94/366 (25.7 per cent) were apparently node positive. Taking a simple difference of these proportions gives a point estimate of -0.3 per cent for the proportion of patients who had positive axillae but were missed by SNB. This estimate is clearly inadmissible. In this situation, a Bayesian analysis yields interpretable point and interval estimates. We consider the single proportion estimate from the validation phase; the difference between independent proportions from the randomized phase, both unconstrained and constrained to non-negativity; and combined information from the two parts of the study. As well as tail-based and highest posterior density interval estimates, we examine three obvious point estimates, the posterior mean, median and mode. Posterior means and medians are similar for the validation and randomized phases separately and combined, all between 2 and 3 per cent, indicating similarity rather than conflict between the two data sources. PMID- 17133627 TI - Evaluation of 384-well formatted sample preparation technologies for regulated bioanalysis. AB - The capabilities and limitations of 384-well formatted sample preparation technologies applied to regulated bioanalysis were evaluated by developing two assays for the simultaneous quantitation of lopinavir and ritonavir, the active ingredients of Kaletra. One method used liquid-liquid extraction (LLE), and the other used solid-phase extraction (SPE). The steps and apparatuses employed by the two methods covered most of those used for bioanalysis. Briefly, the previously validated 96-well formatted assays were adapted to the 384-format with minor modifications. Because the wells of a 384-well plate are clustered together, cross-contamination between adjacent wells was evaluated critically, along with sensitivity, assay throughput, and ruggedness. Samples (35 microL) containing plasma samples (15 microL), internal standard (10 microL), and sodium carbonate (0.5 M, 10 microL to basify the sample) were placed in a 384-well microtiter plate that may contain saquinavir or amprenavir as contamination markers. For LLE preparation, the samples were placed in a deep 384-well plate (300-microL well volume) and extracted with 150 microL of ethyl acetate. Approximately 50 microL of the extracts were removed from each well after phase separation for analysis. For SPE preparation, the fortified samples were transferred to a 384-formatted SPE plate (C18, 5 mg packing). The extracts were eluted from the plate with basified 2-propanol. The LLE or SPE extracts were dried and reconstituted for column-switching high-performance liquid chromatography with tandem mass spectrometric detection (HPLC/MS/MS). The lower limit of quantitation and the assay range were the same as the 96-well formatted assay. If combined with appropriate automation, sample preparation in the 384 well format would be up to five times more efficient than the 96-well format. PMID- 17133629 TI - A statistical comparison of different family history scores. AB - Family history (FH) scores are used for estimating the familial risk (FR), i.e. the level of risk for a particular disease among members of that family. An FH score is created from reports about the disease status of the relatives in each family. The most commonly used score is the dichotomous score (positive when at least one relative is affected), which does not consider the family size, number of affected relatives nor each relative's risk factor profile. Authors have proposed many other FH scores that overcome these deficiencies by using external expected risks adjusted for important risk factors. We consider the use of FH scores in studies, which investigate risk factors for a disease and where family risk is considered as a confounder, and examine through simulations the performance of a variety of FH scores in controlling the FR status. We also examine performance in predicting true FR status. For both criteria, only small differences were found between most of the FH scores, although the dichotomous score performed the poorest. Since the proportion score (the proportion of first degree relatives of the index who have the disease) is the simplest to calculate, use of this score seems to be justified. PMID- 17133630 TI - Regression models for mixed Poisson and continuous longitudinal data. AB - In this article we develop flexible regression models in two respects to evaluate the influence of the covariate variables on the mixed Poisson and continuous responses and to evaluate how the correlation between Poisson response and continuous response changes over time. A scenario for dealing with regression models of mixed continuous and Poisson responses when the heterogeneous variance and correlation changing over time exist is proposed. Our general approach is first to jointly build marginal model and to check whether the variance and correlation change over time via likelihood ratio test. If the variance and correlation change over time, we will do a suitable data transformation to properly evaluate the influence of the covariates on the mixed responses. The proposed methods are applied to the interstitial cystitis data base (ICDB) cohort study, and we find that the positive correlations significantly change over time, which suggests heterogeneous variances should not be ignored in modelling and inference. PMID- 17133631 TI - Assessment of the Gould-Shih procedure for sample size re-estimation. AB - The power of a clinical trial is partly dependent upon its sample size. With continuous data, the sample size needed to attain a desired power is a function of the within-group standard deviation. An estimate of this standard deviation can be obtained during the trial itself based upon interim data; the estimate is then used to re-estimate the sample size. Gould and Shih proposed a method, based on the EM algorithm, which they claim produces a maximum likelihood estimate of the within-group standard deviation while preserving the blind, and that the estimate is quite satisfactory. However, others have claimed that the method can produce non-unique and/or severe underestimates of the true within-group standard deviation. Here the method is thoroughly examined to resolve the conflicting claims and, via simulation, to assess its validity and the properties of its estimates. The results show that the apparent non-uniqueness of the method's estimate is due to an apparently innocuous alteration that Gould and Shih made to the EM algorithm. When this alteration is removed, the method is valid in that it produces the maximum likelihood estimate of the within-group standard deviation (and also of the within-group means). However, the estimate is negatively biased and has a large standard deviation. The simulations show that with a standardized difference of 1 or less, which is typical in most clinical trials, the standard deviation from the combined samples ignoring the groups is a better estimator, despite its obvious positive bias. PMID- 17133632 TI - Synthetic molecular motors and mechanical machines. AB - The widespread use of controlled molecular-level motion in key natural processes suggests that great rewards could come from bridging the gap between the present generation of synthetic molecular systems, which by and large rely upon electronic and chemical effects to carry out their functions, and the machines of the macroscopic world, which utilize the synchronized movements of smaller parts to perform specific tasks. This is a scientific area of great contemporary interest and extraordinary recent growth, yet the notion of molecular-level machines dates back to a time when the ideas surrounding the statistical nature of matter and the laws of thermodynamics were first being formulated. Here we outline the exciting successes in taming molecular-level movement thus far, the underlying principles that all experimental designs must follow, and the early progress made towards utilizing synthetic molecular structures to perform tasks using mechanical motion. We also highlight some of the issues and challenges that still need to be overcome. PMID- 17133633 TI - Gamma-substituted peptide nucleic acids constructed from L-lysine are a versatile scaffold for multifunctional display. PMID- 17133634 TI - Rh-catalyzed enantioselective conjugate addition of arylboronic acids with a dynamic library of chiral tropos phosphorus ligands. AB - A library of 19 chiral tropos phosphorus ligands, based on a free-to-rotate (tropos) biphenol unit and a chiral P-bonded alcohol (11 phosphites, 1-P(O)(2)O to 11-P(O)(2)O) or secondary amine (8 phosphoramidites, 12-P(O)(2)N to 19 P(O)(2)N), were screened, individually and in combinations of two, in the rhodium catalyzed asymmetric conjugate addition of arylboronic acids to enones and enoates. High enantioselectivities (up to 99 % ee) and excellent yields were obtained in the addition to either cyclic or acyclic substrates. The flexible biphenolic P ligands outperformed the analogous rigid binaphtholic P ligands. Variable-temperature (31)P NMR studies revealed that the biphenolic ligands are tropos even at low temperature. Only below 190 K was a coalescence observed; upon further cooling, two atropisomers were detected. The Rh homocomplexes ([Rh(L(a))(2)](+)) were also studied: in general, a single doublet (P-Rh coupling) was observed in the case of the biphenolic phosphite ligands, over the temperature range 380-230 K, demonstrating their tropos nature in the rhodium complexes even at low temperatures. On the other hand, the phosphoramidites showed different behaviors depending on the structure of the ligand and on the nature of the rhodium source. The spectrum at 230 K of the mixture of [Rh(acac)(eth)(2)] (eth=C(2)H(4)) with phosphite 6-P(O)(2)O and phosphoramidite 19-P(O)(2)N (the most enantioselective ligand combination in the conjugate addition reaction) revealed the presence of four homocomplexes (total approximately 40 %: [Rh{6-P(O)(2)O}(2)], [Rh{(aR)-19-P(O)(2)N}(2)], [Rh{(aS)-19 P(O)(2)N}(2)], [Rh{(aR)-19-P(O)(2)N}{(aS)-19-P(O)(2)N}]) and one heterocomplex, [Rh{6-P(O)(2)O}{(aR)-19-P(O)(2)N}] (approximately 60 %) In the heterocomplex, the biphenol-derived phosphite is free to rotate (tropos) while the biphenol-derived phosphoramidite shows a temperature-dependent tropos/atropos behavior (coalescence temperature=310 K). PMID- 17133635 TI - One-electron versus two-electron mechanisms in the oxidative addition reactions of chloroalkanes to amido-bridged rhodium complexes. AB - The compound syn-[{Rh(mu-NH{p-tolyl})(CNtBu)(2)}(2)] (1) oxidatively adds C--Cl bonds of alkyl chlorides (RCl) and dichloromethane to each metal centre to give the cationic complexes syn-[{Rh(mu-NH{p-tolyl})(eta(1)-R)(CNtBu)(2)}(2)(mu-Cl)]Cl and anti-[{Rh(mu-NH{p-tolyl})Cl(CNtBu)(2)}(2)(mu-CH(2))]. Reaction of 1 with the chiral alkyl chloride (-)-(S)-ClCH(Me)CO(2)Me (R*Cl) gave [{Rh(mu-NH{p tolyl})(eta(1)-R*)(CNtBu)(2)}(2)(mu-Cl)]Cl ([3]Cl) as an equimolecular mixture of the meso form (R,S)-[3]Cl-C(s) and one enantiomer of the chiral form [3]Cl-C(2). This reaction, which takes place in two steps, was modeled step-by-step by reacting the mixed-ligand complex syn-[(cod)Rh(mu-NH{p-tolyl})(2)Rh(CNtBu)(2)] (4) with R*Cl, as a replica of the first step, to give [(cod)Rh(mu-NH{p tolyl})(2)RhCl(eta(1)-R*)(CNtBu)(2)] (5) with racemization of the chiral carbon. Further treatment of 5 with CNtBu to give the intermediate [(CNtBu)(2)Rh(mu-NH{p tolyl})(2)RhCl(eta(1)-R*)(CNtBu)(2)], followed by reaction with R*Cl reproduced the regioselectivity of the second step to give (R,S)-[3]Cl-C(s) and [3]Cl-C(2) in a 1:1 molar ratio. Support for an S(N)2 type of reaction with inversion of the configuration in the second step was obtained from a similar sequence of reactions of 4 with ClCH(2)CO(2)Me first, then with CNtBu, and finally with R*Cl to give [(CNtBu)(2)(eta(1)-CH(2)R)Rh(mu-NH{p-tolyl})(2)(mu-Cl)Rh(eta(1) R*)(CNtBu)(2)]Cl (R = CO(2)Me, [7]Cl) as a single enantiomer with the R configuration at the chiral carbon. The reactions of 1 with (+)-(S) XCH(2)CH(CH(3))CH(2)CH(3) (X = Br, I) gave the related complexes [{Rh(mu-NH{p tolyl})(eta(1)-CH(2)CH(CH(3))CH(2)CH(3))(CNtBu)(2)}(2)(mu-X)]X, probably by following an S(N)2 profile in both steps. PMID- 17133636 TI - An unexpected new optimum in the structure space of DNA solubilizing single walled carbon nanotubes. AB - Here we report quantitative data on the amount of single-walled carbon nanotubes that can be suspended with oligodeoxynucleotides in aqueous buffer, together with rate constants for the thermal denaturation of the resulting DNA-nanotube complexes at elevated temperatures. Sequence motifs d(GT)n and d(AC)n with n=2, 3, 5, 10, 20, or 40 were employed, both individually and as equimolar mixtures of the complementary strands. Unexpectedly, the greatest suspending efficiency was found for the mixture of short, complementary oligonucleotides d(GT)3 and d(AC)3. Unlike the suspending efficiency, the kinetic stability of the nanotube suspensions increases with increasing chain length of the DNA, with half life times of >25 h at 90 degrees C for the complexes of the longest strands. Our results identify a new, unexpected optimum in DNA sequence space for suspending carbon nanotubes. They also demonstrate that suspending power depends on the presence of complementary strands. Exploratory assays suggest that nanotubes can be deposited site-selectively from suspensions formed with short DNA sequences. PMID- 17133637 TI - Tuning of the structures of chiral phosphane-phosphites: application to the highly enantioselective synthesis of alpha-acyloxy phosphonates by catalytic hydrogenation. AB - A family of new chiral phosphane-phosphites 5 has been prepared and employed in the synthesis of rhodium complexes of formulation [Rh(cod)(5)]BF4 (7). The use of bulky phosphane or phosphite groups in the preparation of 7 avoids the formation of undesired disubstituted complexes, one of which (9 a) has been isolated and characterized. Ligands 5 display important differences from the bulkier phosphane phosphites 1: complexes 7-unlike their rigid [Rh(cod)(1)]BF4 counterparts-show fluxional behaviour in solution, consistent with backbone oscillation around the coordination plane. A detailed screening of ligands 1 and 5 in catalytic asymmetric hydrogenations of enol phosphonates 12 demonstrated a critical influence of the steric characteristics of the phosphane-phosphite in the course of the reaction, and optimization of the two phosphorus functionalities resulted in the production of versatile and efficient catalysts for this class of hydrogenations: enantioselectivities of up to 98% ee were thus obtained with substrates bearing an alkyl substituent in the beta-position, while for their challenging aryl counterparts values of up to 92% ee were achieved. The coordination mode of phosphonate 12 a towards a Rh phosphane-phosphite fragment has also been investigated and a preference of the olefin fragment to occupy the position cis to the phosphite group has been observed. From this observation an interpretation of the configurations of the hydrogenated phosphonates has also been made. PMID- 17133638 TI - Synthetic, structural, and theoretical investigations of alkali metal germanium hydrides--contact molecules and separated ions. AB - The preparation of a series of crown ether ligated alkali metal (M=K, Rb, Cs) germyl derivatives M(crown ether)nGeH3 through the hydrolysis of the respective tris(trimethylsilyl)germanides is reported. Depending on the alkali metal and the crown ether diameter, the hydrides display either contact molecules or separated ions in the solid state, providing a unique structural insight into the geometry of the obscure GeH3- ion. Germyl derivatives displaying M--Ge bonds in the solid state are of the general formula [M([18]crown-6)(thf)GeH3] with M=K (1) and M=Rb (4). The compounds display an unexpected geometry with two of the GeH3 hydrogen atoms closely approaching the metal center, resulting in a partially inverted structure. Interestingly, the lone pair at germanium is not pointed towards the alkali metal, rather two of the three hydrides are approaching the alkali metal center to display M--H interactions. Separated ions display alkali metal cations bound to two crown ethers in a sandwich-type arrangement and non-coordinated GeH3 ions to afford complexes of the type [M(crown ether)2][GeH3] with M=K, crown ether=[15]crown-5 (2); M=K, crown ether=[12]crown-4 (3); and M=Cs, crown ether=[18]crown-6 (5). The highly reactive germyl derivatives were characterized by using X-ray crystallography, 1H and 13C NMR, and IR spectroscopy. Density functional theory (DFT) and second-order Moller-Plesset perturbation theory (MP2) calculations were performed to analyze the geometry of the GeH3- ion in the contact molecules 1 and 4. PMID- 17133639 TI - One-step fabrication and high photocatalytic activity of porous TiO2 hollow aggregates by using a low-temperature hydrothermal method without templates. AB - Porous TiO2 hollow aggregates have been synthesized on a large scale by means of a simple hydrothermal method without using any templates. The as-prepared products were characterized by means of field emission scanning electron microscopy, XRD, TEM, nitrogen adsorption, UV/Vis diffuse reflectance spectroscopy, and FTIR spectroscopy. The photocatalytic activity of the aggregates was demonstrated through the photocatalytic degradation of Rhodamine B. Structural characterization indicates that the porous TiO2 aggregates are 500 800 nm in diameter and display mesoporous structure. The average pore sizes and BET surface areas of the aggregates are 12 nm and 168 m2 g-1, respectively. Optical adsorption investigations show that the aggregates possess an optical band-gap energy of 3.36 eV. The as-prepared products were substantially more effective photocatalysts than the commercially available photocatalyst P25. The dye degradation rate of the porous TiO2 hollow aggregates is more than twice that of P25. The high photoactivities of the aggregates can be attributed to the combined effects of several factors, namely, large surface areas, the existence of mesopores, and the high band-gap energy. In addition, the as-prepared products can be easily recycled. PMID- 17133640 TI - Coordination chemistry of a kinetically stabilized germabenzene: syntheses and properties of stable eta6-germabenzene complexes coordinated to transition metals. AB - The first stable eta6-germabenzene complexes, that is, [M(CO)3(eta6-C5H5GeTbt)] {M=Cr (2), Mo (3), and W (4); Tbt=2,4,6-tris[bis(trimethylsilyl)methyl]phenyl}, have been synthesized by ligand-exchange reactions between [M(CO)3(CH3CN)3] (M=Cr, Mo, and W) and the kinetically stabilized germabenzene 1 and characterized by 1H and 13C NMR, IR, and UV/Vis spectroscopy. In the 1H and 13C NMR spectra of 2-4, all of the signals for the germabenzene rings were shifted upfield relative to their counterparts in the free germabenzene 1. X-ray crystallographic analysis of 2 and 4 revealed that the germabenzene ligand was nearly planar and was coordinated to the M(CO)3 group (M=Cr, W) in an eta6 fashion. The formation of complexes 2-4 from germabenzene 1 should be noted as the application of germaaromatics as 6pi-electron ligands toward complexation with Group 6 metals. On the other hand, treatment of 1 with [{RuCp*Cl}4] (Cp*=C5Me5) in THF afforded a novel eta5-germacyclohexadienido complex of ruthenium-[RuCp*{eta5-C5H5GeTbt(Cl)}] (9)-instead of the expected eta6-germabenzene-ruthenium cationic complex [RuCp*{eta6-C5H5GeTbt}]Cl (10). Crystallographic structural analysis of 9 showed that the five carbon atoms of the germacyclohexadienido ligand of 9 were coordinated to the Ru center in an eta5 fashion. PMID- 17133641 TI - Oxidation of linear trinuclear ruthenium complexes [Ru(3)(dpa)(4)Cl(2)] and [Ru(3)(dpa)(4)(CN)(2)]: synthesis, structures, electrochemical and magnetic properties. AB - The neutral, monocationic, and dicationic linear trinuclear ruthenium compounds [Ru(3)(dpa)(4)(CN)(2)], [Ru(3)(dpa)(4)(CN)(2)][BF(4)], [Ru(3)(dpa)(4)Cl(2)][BF(4)], and [Ru(3)(dpa)(4)Cl(2)][BF(4)](2) (dpa=the anion of dipyridylamine) have been synthesized and characterized by various spectroscopic techniques. Cyclic voltammetric and spectroelectrochemical studies on the neutral and oxidized compounds are reported. These compounds undergo three successive metal-centered one-electron-transfer processes. X-ray structural studies reveal a symmetrical Ru(3) unit for these compounds. While the metal--metal bond lengths change only slightly, the metal--axial ligand lengths exhibit a significant decrease upon oxidation of the neutral complex. The electronic configuration of the Ru(3) unit changes as the axial chloride ligands are replaced by the stronger "pi-acid" cyanide axial ligands. Magnetic measurements and (1)H NMR spectra indicate that [Ru(3)(dpa)(4)Cl(2)] and [Ru(3)(dpa)(4)Cl(2)][BF(4)](2) are in a spin state of S=0 and [Ru(3)(dpa)(4)Cl(2)][BF(4)], [Ru(3)(dpa)(4)(CN)(2)], and [Ru(3)(dpa)(4)(CN)(2)][BF(4)] are in spin states of S=1/2, 1, and 3/2, respectively. These results are consistent with molecular orbital (MO) calculations. PMID- 17133642 TI - Synthesis and antigenic analysis of the BclA glycoprotein oligosaccharide from the Bacillus anthracis exosporium. AB - The glycoprotein BclA is an important constituent of the exosporium of Bacillus anthracis spores. This glycoprotein is substituted with an oligosaccharide composed of a beta-L-rhamnoside substituted with the previously unknown terminal saccharide, 2-O-methyl-4-(3-hydroxy-3-methylbutanamido)-4,6-dideoxy-D glucopyranose, also referred to as anthrose. Anthrose has not been found in spores of B. cereus and B. thuringiensis, making it a potential species-specific marker for B. anthracis. In order to study the antigenicity of anthrose, efficient syntheses of an anthrose-containing trisaccharide and a series of structurally related analogues were developed. The analogues lacked either the methyl ether at C-2 or contained modified C-4 amino functionalities of anthrose. The synthetic compounds were equipped with an aminopropyl spacer to facilitate conjugation to the carrier proteins mariculture Keyhole Limpet Hemocyanin (mcKLH) and bovine serum albumin (BSA). Serum antibodies of rabbits immunized with live or irradiated spores of B. anthracis Sterne 34F(2) were able to recognize the synthetic trisaccharide-mcKLH conjugate. The specificity of the interaction was confirmed by competitive inhibition with the free- and BSA-conjugated trisaccharides. Inhibition using the trisaccharide analogues demonstrated that the isovaleric acid moiety of anthrose is an important structural motif for antibody recognition. These data demonstrate that 1) anthrose is a specific antigenic determinant of the B. anthracis Sterne spore; 2) this antigen is presented to the immune system of rabbits receiving the anthrax live-spore vaccine; 3) synthetic analogues of the oligosaccharide retain the antigenic structure; and 4) the antigenic region is localized to specific terminal groups of the oligosaccharide. Collectively these data provide an important proof-of concept step in the synthesis and development of spore-specific reagents for detection and targeting of non-protein structures in B. anthracis. PMID- 17133643 TI - Tetrabromocinnamic acid (TBCA) and related compounds represent a new class of specific protein kinase CK2 inhibitors. AB - Abnormally high constitutive activity of protein kinase CK2, levels of which are elevated in a variety of tumours, is suspected to underlie its pathogenic potential. The most widely employed CK2 inhibitor is 4,5,6,7 tetrabromobenzotriazole (TBB), which exhibits a comparable efficacy toward another kinase, DYRK1 a. Here we describe the development of a new class of CK2 inhibitors, conceptually derived from TBB, which have lost their potency toward DYRK1 a. In particular, tetrabromocinnamic acid (TBCA) inhibits CK2 five times more efficiently than TBB (IC50 values 0.11 and 0.56 microM, respectively), without having any comparable effect on DYRK1 a (IC50 24.5 microM) or on a panel of 28 protein kinases. The usefulness of TBCA for cellular studies has been validated by showing that it reduces the viability of Jurkat cells more efficiently than TBB through enhancement of apoptosis. Collectively taken, the reported data support the view that suitably derivatized tetrabromobenzene molecules may provide powerful reagents for dissecting the cellular functions of CK2 and counteracting its pathogenic potentials. PMID- 17133644 TI - Selective labeling of proteins by using protein farnesyltransferase. AB - The challenging task of identifying and studying protein function has been greatly aided by labeling proteins with reporter groups. Here, we present a strategy that utilizes an enzyme that labels a four-residue sequence appended onto the C terminus of a protein, with an alkyne-containing substrate. By using a bio-orthogonal cycloaddition reaction, a fluorophore that carried an azide moiety was then covalently coupled to the alkyne appended on the protein. FRET was used to calculate a Forster (R) distance of 40 A between the eGFP chromophore and the newly appended Texas Red fluorophore. This experimental value is in good agreement with the predicted R value determined by using molecular modeling. The small recognition tag, the high specificity of the enzyme, and the orthogonal nature of the derivatization reaction will make this approach highly useful in protein chemistry. PMID- 17133645 TI - Learning from directed evolution: Further lessons from theoretical investigations into cooperative mutations in lipase enantioselectivity. AB - An earlier experimental study, which involved the directed evolution of enantioselective lipase variants from Pseudomonas aeruginosa as catalysts in the hydrolytic kinetic resolution of 2-methyl-decanoic acid p-nitrophenyl ester, provided a mutant with six mutations. Consequently, the selectivity factor was found to increase from E = 1.1 for the wild-type to E = 51 for the best mutant. Only one of the amino acid exchanges in this mutant was found to occur next to the binding pocket, the other mutations being remote. Our previous theoretical analysis with molecular-dynamics simulations helped to unveil the source of enhanced enantioselectivity: a relay mechanism that involves two of the six mutations was shown to induce strong cooperativity. In this investigation, single, double, and triple mutants were constructed and tested as enantioselective catalysts. This study supports our original postulate regarding the relay mechanism, offers further mechanistic insight into the role of individual mutations, and provides mutants that display even higher enantioselectivity (E of up to 64). PMID- 17133646 TI - Predicting the nature and timing of epimerisation on a modular polyketide synthase. PMID- 17133647 TI - A boosting approach to flexible semiparametric mixed models. AB - In linear mixed models the influence of covariates is restricted to a strictly parametric form. With the rise of semi- and non-parametric regression also the mixed model has been expanded to allow for additive predictors. The common approach uses the representation of additive models as mixed models. An alternative approach that is proposed in the present paper is likelihood based boosting. Boosting originates in the machine learning community where it has been proposed as a technique to improve classification procedures by combining estimates with reweighted observations. Likelihood based boosting is a general method which may be seen as an extension of L2 boost. In additive mixed models the advantage of boosting techniques in the form of componentwise boosting is that it is suitable for high dimensional settings where many explanatory variables are present. It allows to fit additive models for many covariates with implicit selection of relevant variables and automatic selection of smoothing parameters. Moreover, boosting techniques may be used to incorporate the subject specific variation of smooth influence functions by specifying 'random slopes' on smooth effects. This results in flexible semiparametric mixed models which are appropriate in cases where a simple random intercept is unable to capture the variation of effects across subjects. PMID- 17133648 TI - Pseudoscore-based estimation from biased observations. AB - There are many practical situations where observation of the primary variable Y for individuals in a population is incomplete and depends on some auxiliary variables X that are potentially correlated with Y. We consider parameter estimation for the distribution of Y with the incomplete data, without specifying the underlying association between Y and X. The approach is based on a class of pseudoscore functions using available information of X. We demonstrate the consistency and asymptotic normality of the estimators and study their finite sample properties in various situations via simulation. The methodology is illustrated by an example involving kindergarten readiness skills in children with sickle cell disease. PMID- 17133649 TI - Computerized adaptive testing for measuring development of young children. AB - Developmental indicators that are used for routine measurement in The Netherlands are usually chosen to optimally identify delayed children. Measurements on the majority of children without problems are therefore quite imprecise. This study explores the use of computerized adaptive testing (CAT) to monitor the development of young children. CAT is expected to improve the measurement precision of the instrument. We do two simulation studies - one with real data and one with simulated data - to evaluate the usefulness of CAT. It is shown that CAT selects developmental indicators that maximally match the individual child, so that all children can be measured to the same precision. PMID- 17133650 TI - Quantification of D-amino acids in the central nervous system of Aplysia californica by liquid chromatography/tandem mass spectrometry. AB - A sensitive, specific and reliable liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed for simultaneous determination of D-amino acids in the central nervous system (CNS) of Aplysia californica. In order to correct for any potential matrix effects on measured signals, deuterium-labeled L Asp-d3 was used as an internal standard. Pre-column derivatization of the sample with 7-fluoro-4-nitrobenzoxadiazole (NBD-F) allowed both effective in-line pre concentration and sensitive MS/MS detection of the analytes. An extraction column (50x0.25 mm, 5 microm C18 silica particles) was used to pre-concentrate/stack samples. Enantiomeric separation of amino acid enantiomers was achieved on a chiral column packed with teicoplanin aglycone bonded silica particles (170x0.25 mm, 5 microm) with an MS-friendly mobile phase. The characteristic precursor to product ion transitions, m/z 297-->279 (for NBD-Asp), m/z 269-->223 (For NBD Ser), m/z 311-->293 (for NBD-Glu) and m/z 300-->282 (for NBD-L-Asp-d3) were monitored for the quantification. Samples from the CNS of A. californica and heart tissues were analyzed. D-Asp was detected at high levels in all the ganglia and nerve tissues, but not in the heart tissue. Further, neither D-Ser nor D-Glu was detected in Aplysia, a widely used neuronal model. PMID- 17133651 TI - Adaptive design method based on sum of p-values. AB - Bauer and Kohne proposed an adaptive design using Fisher's combination of independent p-values based on subsamples from different stages (Biometrics 1994; 50(4):1029-1041). Their method provides great flexibility in the selection of statistical methods for hypothesis testing of subsamples. However, the choices for the stopping boundaries are not flexible enough to meet practical needs (Biometrics 2001; 57(3): 886-891). In this paper, an adaptive design method is proposed using linear combination of the independent p-values. The method provides great flexibility in the selection of stopping boundaries and no numerical integration is required for the two-stage designs. The stopping boundaries and p-values can be calculated manually. The operating characteristics of the adaptive designs are studied using computer simulations with and without sample size adjustment. Examples are presented for superiority and non inferiority trials with different endpoints (normal, binary, and survival) under different adaptations. The statistical efficiency of the proposed method is compared with other methods based on conditional power. PMID- 17133652 TI - Trends in suicide rates of the elderly in Austria, 1970-2004: an analysis of changes in terms of age groups, suicide methods and gender. AB - BACKGROUND: Suicides of the elderly (persons aged 65 and older) make up a large proportion of total suicides. Since suicide rates of the elderly are highest in western populations, addressing them as a risk group in prevention plans has been recommended. In order to assess possible approaches to prevention strategies, this study examines high-risk groups of the elderly. METHODS: We examined official statistics on suicides that occurred in Austria between 1970-2004 (18,101 Suicides of the elderly). We analyzed time trends and differences in suicide methods as well as in age groups and both genders of the elderly. RESULTS: Three major high-risk groups were identified: elderly male suicides by firearms; elderly female suicides by poisoning, which occur more often with increasing age; and suicides of both genders by jumping from heights. CONCLUSION: Besides conducting treatment of psychiatric disorders of the elderly, restricting the means to commit suicide may help to prevent it among the elderly. Such specific prevention strategies should be implemented in national suicide prevention plans for the high-risk groups identified in this study. PMID- 17133653 TI - Potential cytotoxic activity of some Brazilian seaweeds on human melanoma cells. AB - In vitro screening of the crude extracts of some Brazilian coastal seaweeds for cytotoxic activity against a cultured human melanoma cancer cell line using the sulphorhodamine B assay was performed. The crude dichloromethane:chloroform extract of Stypopodium zonale showed good cytotoxic activity against the C32 cell line. The crude acetone extract and aqueous phase of Lobophora variegata did not show any activity, but semi-purified fractions XAD LOB I and II could inhibit the growth of melanoma cells. The crude acetone extract of Caulerpa racemosa showed some cytotoxicity, but caulerpin isolated from this extract did not show any such activity. The crude acetone extract of Spatoglossum schroederi was not able to inhibit the growth of C32 melanoma cells. PMID- 17133654 TI - Stronger symptoms of depression predict high coronary heart disease mortality in older men and women. AB - BACKGROUND: Most cohort studies have found depressive symptoms to be associated with increased cardiovascular mortality in the elderly, but follow-up times have often been short and study populations small. OBJECTIVES: To describe associations between stronger symptoms of depression and the risk of death from coronary heart disease (CHD) or myocardial infarction (MI) in elderly Finnish subjects free of CHD at the baseline. METHODS: This study is a prospective population-based epidemiological and clinical twelve-year follow-up study in Lieto Health Centre, Finland. The basic population consisted of 1196 elderly (64 years of age or older) persons who lived in the municipality of Lieto in southwestern Finland in 1990. The occurrence of CHD was determined on the basis of electrocardiographic (ECG) findings, Rose questionnaire and the diagnoses in medical records. The persons with CHD were excluded from the study population. Symptoms of depression at the baseline were measured with the Zung Self-rating Depression Scale (ZSDS). Mortality was followed up for about 12 years. RESULTS: The Kaplan-Meier survival curves showed stronger symptoms of depression to be related to high risks of mortality from CHD or MI among men and women. According to the Cox model for men significant predictors for higher risk of CHD or MI mortality were stronger symptoms of depression, higher age and a large number of medications in use. When women were examined, significant predictors were stronger symptoms of depression and a large number of medications in use. CONCLUSIONS: Stronger symptoms of depression are an independent risk factor for high CHD or MI mortality in aged Finnish men and women free of CHD at baseline. PMID- 17133655 TI - Evaluating the outcome of interventions on quality of life in dementia: selection of the appropriate scale. AB - BACKGROUND: A literature study was conducted to contribute to an adequate use of quality of life (QoL) instruments for the evaluation of interventions in dementia care by providing an overview of properties of QoL measures that acknowledge domains important to dementia patients. METHODS: Domains important to patients, and domains that professional caregivers in different settings focus on, are compared to domains represented in nine QoL instruments. Data on psychometrics and applicability are generated. RESULTS: Four instruments best represent domains of QoL important to patients and domains professional caregivers in 24-h care and daytime activities focus on. Two are self-rating instruments: Schedule for the Evaluation of Individual Quality of Life, applicable in mild dementia, measuring individual QoL of patient and informal caregiver, and Quality of Life-Alzheimer's Disease Scale, which can be applied up to moderately severe dementia. For patients with advanced dementia receiving residential care, the observational instruments Qualidem and Discomfort scale-Dementia of Alzheimer Type are recommended. The first is easily administered by nurse assistants or occupational therapists and covers several QoL domains on which they focus in daily practice. The second can be used by various professionals. CONCLUSIONS: QoL assessment provides a format for patients and (in)formal caregivers to express whether an intervention has made an important difference to the patient's life. Improvement of QoL in dementia should have high priority in care, treatment and research. This study shows that severity of dementia, care type, setting, and the specific QoL domains an intervention focuses on, determine which QoL instrument is most appropriate in a specific situation. PMID- 17133656 TI - Depression in veterans with Parkinson's disease: frequency, co-morbidity, and healthcare utilization. AB - OBJECTIVE: To determine the frequency of depression in Parkinson's disease (PD) in routine clinical care, and to examine its association with co-morbid psychiatric and medical conditions and healthcare utilization. METHODS: Depression diagnoses and healthcare utilization data for all male veterans with PD age 55 or older seen in fiscal year 2002 (n = 41,162) were analyzed using Department of Veterans Affairs (VA) national databases. Frequencies of co-morbid disorders and healthcare utilization were determined for depressed and non depressed patients; associations with depression were examined using multivariate logistic regression models. RESULTS: A depression diagnosis was recorded for 18.5% of PD patients, including major depression in 3.9%. Depression decreased in frequency and severity with increasing age. In multivariate logistic regression models, depressed patients had significantly greater psychiatric and medical co morbidity, including dementia, psychosis, stroke, congestive heart failure, diabetes, and chronic obstructive pulmonary disease than non-depressed patients (all p < 0.01). Depressed PD patients were also significantly more likely to have medical (OR = 1.34, 95% CI = 1.25-1.44) and psychiatric hospitalizations (OR = 2.14, 95% CI = 1.83-2.51), and had more outpatient visits (p < 0.01), than non depressed PD patients in adjusted models. CONCLUSION: Depression in PD in non tertiary care settings may not be as common or as severe as that seen in specialty care, though these findings also may reflect under-recognition or diagnostic imprecision. The occurrence of depression in PD is associated with greater psychiatric and medical co-morbidity, and greater healthcare utilization. These findings suggest that screening for depression in PD is important and should be embedded in a comprehensive psychiatric, neuropsychological, and medical evaluation. PMID- 17133657 TI - Diabetes and depression, a circular connection. PMID- 17133658 TI - Ask the doctor. I have heard that coconut is bad for the heart and that it is good for the heart. Which is right? PMID- 17133659 TI - When expectation fails. PMID- 17133660 TI - Something to see. PMID- 17133661 TI - Northern exposure. PMID- 17133662 TI - Made-to-measure postdocs. PMID- 17133663 TI - Abstracts from the Rehabilitative Ultrasound Imaging Symposium. May 8-10, 2006. San Antonio, Texas, USA. PMID- 17133664 TI - Surgical innovation: too risky to remain unregulated? PMID- 17133665 TI - Current controversies in pharmacoepidemiology. Proceedings of a symposium. May 27 28, 2005. Kolding, Denmark. PMID- 17133666 TI - [Pulmonary hypertension, the "bete noire" of systemic diseases: from systemic sclerosis to hereditary hemorrhagic telangiectasia]. PMID- 17133667 TI - Hypertension in pregnancy: a Russian initiative. PMID- 17133668 TI - History of cardiology: Paul Louis Duroziez, MD. PMID- 17133669 TI - [Modern diagnostic assessment of the upper urinary tract using multislice CT urography]. AB - The advent of Multislice Computed Tomography (MSCT) has made evaluation of the entire urinary tract with high-resolution sections during a single breath-hold a reality. Acquisition of multiple thin overlapping slices provides excellent two dimensional (2D) and three-dimensional (3D) reformations of the urinary tract. The concept of "Multislice CT Urography (MSCTU)" has emerged from this technical improvement. As a result, a wide range of pathologies inside and outside the urinary tract can be identified. During the last several years, MSCTU has challenged intravenous urography (IVU) in the evaluation of urinary tract abnormalities. Compared with IVU, MSCT(U) is more sensitive and specific in the detection and characterization of a variety of urinary tract disorders, including renal masses and urolithiasis. The main advantage of IVU has been its ability to offer excellent delineation of pelvicalyceal and ureteral anatomy and to depict subtle uroepithelial abnormalities. MSCTU has already shown promising results for overcoming this challenge. Optimal opacification and distension appear to be an essential requirement for a thorough evaluation of the collecting system. Dedicated preparation strategies have been developed to meet these technical difficulties. The biggest disadvantage of MSCTU is the significant radiation exposure. For broad routine clinical application, there is still a need for dose reduction protocols despite the ongoing technical developments in MSCTU. In this article, we outline the different concepts of technical processing for MSCTU and summarize the current role of MSCTU in the evaluation of the upper urinary tract. PMID- 17133670 TI - [MRI for the detection of anorectal fistulas]. AB - MRI is accepted as one of the major diagnostic tools for the detection of anorectal fistulas and abscesses. Noninvasiveness and high accuracy are advantageous hallmarks of this technique. In general, the purpose of imaging anal fistulas is to reduce the risk of recurrence, incontinence and non-healing. To achieve these goals, the applied method must provide the proctologist with detailed information. In this context, MRI acts as a guide for surgeons to accurately plan fistula operations. Another aspect is the follow-up of conservatively treated patients with fistulizing Crohn's disease. In 2000, subtraction MR fistulography was introduced as new imaging technique. This review provides an overview of the entire spectrum of diagnostic modalities for anorectal fistulas with emphasis on subtraction MR fistulography. PMID- 17133671 TI - [(Modic) signal alterations of vertebral endplates and their correlation to a minimally invasive treatment of lumbar disc herniation using epidural injections]. AB - PURPOSE: To study the influence of (Modic) signal alterations (SA) of the cartilage endplate (CEP) of vertebrae L3-S1 on the outcome of an in-patient minimally invasive treatment (MIT) using epidural injections on patients with lumbar disc herniation (LDH). MATERIALS AND METHODS: The MR images of 59 consecutive patients with LDH within segments L3/L4 - L5/S1 undergoing in-patient minimally invasive treatment with epidural injections were evaluated in a clinical study. The (Modic) signal alterations of the CEP were recorded using T1- and T2-weighted sagittal images. On the basis of the T2-weighted sagittal images, the extension and distribution of the SA were measured by dividing each CEP into 9 areas. The outcome of the MIT was recorded using the Oswestry Disability Index (ODI) before and after therapy and in a 3-month follow-up. Within a subgroup of patients (n = 35), the distribution and extension of the signal alterations were correlated with the development of the ODI. RESULTS: Segments with LDH showed significantly more (p < 0.001) SA of the CEP than segments without LDH. Although the extension of the SA was not dependent on sex, it did increase significantly with age (p = 0.017). The outcome after MIT did not depend on the sex and age of the patients nor on the type of LDH. The SA extension tended to have a negative correlation with the outcome after MIT after 3 months (p = 0.071). A significant negative correlation could be established between the SA extension in the central section of the upper endplate and the outcome after 3 months (p = 0.019). CONCLUSION: 1. Lumbar disc herniation is clearly associated with the prevalence of (Modic) signal alterations. 2. Extensive signal alterations tend to correlate with a negative outcome of an MIT using epidural injections. 3. Such SA in the central portion of the upper CEP correlate significantly with a negative treatment result. 4. The central portion of the upper CEP being extensively affected by (Modic) SA is a negative predictor for the success of a minimally invasive pain therapy. PMID- 17133672 TI - [Diagnostic imaging of enthesitis: a new field for radiologists?]. PMID- 17133673 TI - Re: Why omega-3 fatty acids are important to neurosurgeons (Ausman JI, Surg Neurol 2006;65:325). Omega-3 fatty acids (fish oil) as an anti-inflammatory: an alternative to NSAIDs for discogenic pain (Maroon JC and Bost JW, Surg Neurol 2006;65:326-31). PMID- 17133674 TI - Back to the future. PMID- 17133675 TI - Proceedings and abstracts of the 1st Annual Meeting of the European Diabetes Epidemiology Group of the European Association for the Study of Diabetes, Krakow, Poland, May 20-23, 2006. PMID- 17133676 TI - Proceedings of the 1st Cambridge Conference on Advances in Treating Metastatic Bone Cancer, October 28-29, 2005, Cambridge, Massachusetts, USA. PMID- 17133677 TI - Mind the gaps. PMID- 17133678 TI - Unsure start. PMID- 17133679 TI - Mental wealth. Interview by Lynne Friedli. PMID- 17133680 TI - Tell me a story. PMID- 17133681 TI - Learning together. PMID- 17133682 TI - Outside the box. PMID- 17133683 TI - The bigger picture. PMID- 17133684 TI - Welfare writes. PMID- 17133685 TI - Doing better by depression. PMID- 17133686 TI - Inhuman abuses. PMID- 17133687 TI - [Two new standard vaccinations for infants and small children]. PMID- 17133688 TI - [Nitrous oxide sedation]. PMID- 17133689 TI - A comparison of the postdeployment hospitalization experience of U.S. military personnel following service in the 1991 Gulf War, Southwest Asia after the Gulf War, and Bosnia. AB - Much attention has been given to the impact of deployment on the health of veterans from the 1991 Gulf War. Whereas increases in self-reported symptoms have been common, no specific exposures have been implicated. Some have suggested that stress from deployment is the chief cause for multisymptom conditions among Gulf War veterans, but comparisons with the health of other recent deployers have not been made. We sought to examine the impact of several large military deployments on hospitalization experience. Hospitalization records were examined for all active duty personnel deployed exclusively to the Gulf War, Southwest Asia after the Gulf War, or Bosnia. Cox's hazard modeling was used to assess time until first post-deployment hospitalization, separation from active duty, or December 31, 2000, whichever occurred first, while controlling for influential covariates and temporal changes. Personnel deployed to Southwest Asia after the 1991 Gulf War were at a slight increased risk for any-cause hospitalization and for 3 of the 14 major diagnostic categories when compared with veterans of the 1991 Gulf War. Personnel deployed to Bosnia were at a decreased risk for any-cause hospitalization and 12 of the 14 major diagnostic categories when compared with Gulf War veterans. These findings do not fully explain the complexity of postdeployment health experiences. Although the risk for hospitalization may be associated with regional deployment, it is unlikely that Gulf War veterans are at greater risk of hospitalization due to a specific exposure-related disease. PMID- 17133690 TI - Correlation between airflow patterns and performance of a laboratory fume hood. AB - To understand the physical mechanisms of the contaminant dispersion and containment leakage during the ventilation process through a laboratory fume hood, the complicated three-dimensional flow patterns and the real-time tracer gas (SF6) leakage were studied via the laser-assisted flow visualization method and the standard/special gas sampling technique, respectively. Through flow visualization, the large-scale vortex structures and boundary layer separations were found around the side poles and doorsill of the hood. In the near-wake region of the manikin, large recirculation zones and wavy flow structures were also identified. When tracer gas concentration measurements were conducted point by-point across the sash opening, the areas near the doorsill, the lower parts of the side poles, and the sides of the manikin showed significant contaminant leaks. These areas with high contaminant leaks exactly corresponded to where the flow recirculated or separated. However, when the ANSI/ASHRAE 110-1995 protocol was used to measure the concentration of SF6 at the breathing zone of the manikin, no appreciable leakage was detected. It is suggested that a method based on the aerodynamic features and multipoint leakage detections would reflect a more realistic evaluation of overall performance of laboratory fume hood than a single-point sampling method at the manikin's breathing zone. PMID- 17133691 TI - A preliminary study on gram-negative bacteria (GNB) and their endotoxins in a gin house in India. AB - The presence of byssinosis, an occupational disease found among cotton mill workers, has been well documented in different parts of the world. The disease develops due to exposure to environmental cotton dust. Evidence suggests that the causative agent for the disease is gram-negative bacteria (GNB) and their endotoxins present on the cotton fibers. An investigation was carried out in a gin house in western India. Environmental dust samples were collected by vertical elutriator (VE). Airborne dust concentrations were very high in the working environment: 2.11 mg/m3 in ginning and 0.95 mg/m3 in the press department (p < 0.05), which was higher than the threshold limit value collected by VE (0.2 mg/m3), and higher than the permissible exposure limit for respirable dust (0.5 mg/m3 for nontextile industries using cotton). In the office control site, the dust concentration was 0.31 mg/m3. The Occupational Safety and Health Administration's cotton dust standard permissible exposure limit for respirable dust is 0.2 mg/m3 in yarn manufacturing, 0.75 mg/m3 in slashing and weaving, and 0.5 mg/m3 in nontextile industries using cotton. These samples also showed high concentrations of airborne endotoxin (p < 0.001) in ginning and pressing (2.77 and 1.52 micro g/m3) compared with the office control site (0.009 micro g/m3 measured by Limulus amoebocyte lysate technique). Total enumeration of airborne GNB was carried out qualitatively by the petri plate exposure method and quantitatively by an Andersen 6-stage viable sampler and VE. GNB were recovered in quite high numbers. Among all the GNB, Enterobacter agglomerans were the dominant bacterial flora. Results indicate that gin workers are occupationally exposed to airborne GNB and endotoxins, and require masks. PMID- 17133692 TI - When and why do experts perform exposure measurements? An exploratory study of safety engineers, work environment inspectors, and occupational hygienists. AB - The aim of this study was to describe experts' decision processes leading to measurements of occupational chemical exposure. Safety engineers, clinical occupational hygienists, and work environment inspectors (four from each group) were interviewed according to a semistructured scheme. We analyzed: (i) perceived triggers for measurements, (ii) the experts' decision criteria for performing measurements when there was a trigger, and (iii) sampling strategy. Regarding triggers, all safety engineers reported a "request" from, for example, an employer; all work environment inspectors reported "legal demands;" and all clinical occupational hygienists reported "symptoms among workers." As a decision criterion, all the interviewees reported that they measured only if they expected sufficiently high levels. The results of the present study highlight factors that trigger measurements and are of importance in determining whether measurements will be performed. These factors appear to be related to the expert's professional role and may bias the assessment of exposure. Thus, when using data from routine measurements done by experts, the possibility of a bias needs to be considered. PMID- 17133693 TI - Impact of silanol surface density on the toxicity of silica aerosols measured by erythrocyte haemolysis. AB - Exposures to silica-containing dusts are associated with a risk of developing life-threatening lung diseases. However, the mechanism of silica toxicity is poorly understood. In this work the atomic structure of the surfaces of different silica polymorphs was determined, and a relationship with in vitro silica toxicity was examined. The density of geminal and single silanol groups was quantitatively estimated for different silica polymorphs using a novel molecular modeling method. An association was found between the reported haemolytic activity and modeled densities of surface geminal (but not single) silanol groups on several silica polymorphs. These findings suggest a new view of aerosol toxicity based on the estimation of surface site densities. The results can be used in the development of new toxicological assays for respirable particulates, including nanomaterials. PMID- 17133694 TI - Knowledge of chemical workers on the Occupational Disease Prevention and Control Act of the People's Republic of China. PMID- 17133695 TI - Medicare program; hospital outpatient prospective payment system and CY 2007 payment rates; CY 2007 update to the ambulatory surgical center covered procedures list; Medicare administrative contractors; and reporting hospital quality data for FY 2008 inpatient prospective payment system annual payment update program--HCAHPS survey, SCIP, and mortality. Final rule with comment period and final rule. AB - This final rule with comment period revises the Medicare hospital outpatient prospective payment system to implement applicable statutory requirements and changes arising from our continuing experience with this system, and to implement certain related provisions of the Medicare Prescription Drug, Improvement, and Modernization Act (MMA) of 2003 and the Deficit Reduction Act (DRA) of 2005. In this final rule with comment period, we describe changes to the amounts and factors used to determine the payment rates for Medicare hospital outpatient services paid under the prospective payment system. These changes are applicable to services furnished on or after January 1, 2007. In addition, this final rule with comment period implements future CY 2009 required reporting on quality measures for hospital outpatient services paid under the prospective payment system. This final rule with comment period revises the current list of procedures that are covered when furnished in a Medicare-approved ambulatory surgical center (ASC), which are applicable to services furnished on or after January 1, 2007. This final rule with comment period revises the emergency medical screening requirements for critical access hospitals (CAHs). This final rule with comment period supports implementation of a restructuring of the contracting entities responsibilities and functions that support the adjudication of Medicare fee-for-service (FFS) claims. This restructuring is directed by section 1874A of the Act, as added by section 911 of the MMA. The prior separate Medicare intermediary and Medicare carrier contracting authorities under Title XVIII of the Act have been replaced with the Medicare Administrative Contractor (MAC) authority. This final rule continues to implement the requirements of the DRA that require that we expand the "starter set" of 10 quality measures that we used in FY 2005 and FY 2006 for the hospital inpatient prospective payment system (IPPS) Reporting Hospital Quality Data for the Annual Payment Update (RHQDAPU) program. We began to adopt expanded measures effective for payments beginning in FY 2007. In this rule, we are finalizing additional quality measures for the expanded set of measures for FY 2008 payment purposes. These measures include the HCAHPS survey, as well as Surgical Care Improvement Project (SCIP, formerly Surgical Infection Prevention (SIP)), and Mortality quality measures. PMID- 17133696 TI - Medicare and Medicaid programs; hospital conditions of participation: requirements for history and physical examinations; authentication of verbal orders; securing medications; and postanesthesia evaluations. Final rule. AB - In this rule, we finalize changes to four of the current requirements (or conditions of participation (CoPs)) that hospitals must meet to participate in the Medicare and Medicaid programs. Specifically, this final rule revises and updates our CoP requirements for: Completion of the history and physical examination in the medical staff and the medical record services CoPs; authentication of verbal orders in the nursing service and the medical record services CoPs; securing medications in the pharmaceutical services CoP; and completion of the postanesthesia evaluation in the anesthesia services CoP. We also respond to timely public comments submitted on the proposed rule published in the March 25, 2005 Federal Register (70 FR 15266). The changes specified in this final rule are consistent with current medical practice and will reduce the regulatory burden on hospitals. PMID- 17133697 TI - Medicare program; notification of hospital discharge appeal rights. Final rule. AB - This final rule sets forth requirements for how hospitals must notify Medicare beneficiaries who are hospital inpatients about their hospital discharge rights. Notice is required both for original Medicare beneficiaries and for beneficiaries enrolled in Medicare Advantage (MA) plans and other Medicare health plans subject to the MA regulations. (For purposes of this preamble, these entities will collectively be known as "Medicare health plans"). Hospitals will use a revised version of the Important Message from Medicare (IM), an existing statutorily required notice, to explain the discharge rights. Hospitals must issue the IM within 2 days of admission, and must obtain the signature of the beneficiary or his or her representative. Hospitals will also deliver a copy of the signed notice prior to discharge, but not more than 2 days before the discharge. For beneficiaries who request an appeal, the hospital will deliver a more detailed notice. PMID- 17133698 TI - Gorilla conservation project takes 'one-health' approach. Benefits extend beyond endangered apes. PMID- 17133699 TI - Health insurance is expensive. Not being insured can be more costly. PMID- 17133700 TI - Bringing more bovine practitioners into the bold. The AABP's new foundation a relevant resource. PMID- 17133701 TI - Hatcher advocates more time on farm. Preparing practitioners for front line a priority for AABP president. PMID- 17133702 TI - Animals rights issues continue to generate discussion. PMID- 17133703 TI - EPSDT: Medicaid's critical but controversial benefits program for children. AB - The Early and Periodic Screening, Diagnosis, and Treatment (EPSDT) program under Medicaid provides the most comprehensive set of health benefits for children and adolescents in the public or private sector. A cornerstone of early childhood preventive and treatment services in the nation's health care "safety net," the EPSDT program serves nearly 30 million low-income children, including children with disabilities and special needs. Over the years, states have expressed frustration with the administrative burdens of EPSDT requirements. Rising Medicaid costs have put all Medicaid benefits, including the EPSDT program, in the budgetary crosshairs. This issue brief reviews the fundamental characteristics of the EPSDT program and highlights some of the challenges it has faced over they ears. This paper also describes some of the changes proposed to preserve access to comprehensive care while controlling costs and encouraging administrative simplification and flexibility. PMID- 17133704 TI - [Phenotypic analysis of luxS gene deletion mutants and its application in virulence regulation research in group B Streptococcus]. AB - OBJECTIVE: To explore the mechanism of virulence regulation in group B streptococcus (BGS) by studying LuxS-related AI-2 quorum-sensing pathway in GBS. METHODS: luxS gene deletion mutants of GBS (Delta lusX) were characterized by reverse transcription (RT)-PCR, colony immunoblot analysis, growth curve measurement, and cAMP determination. Functional analysis of luxS in the deletion mutants was conducted by bioluminescence assay. RESULTS: Genetic analysis results showed that the luxS deletion in the mutant 515-Delta lusX caused upregulation of scpB gene expression. Phenotypic analysis revealed that, in comparison with the wild strain, 515-Delta lusX mutant grew slowly in DCM media but quickly in THY media. An approximately two-fold decrement in bioluminescence was detected in the luxS deletion mutants as compared with the wild strain. CONCLUSION: This study confirms the importance of LuxS molecule in the AI-2 quorum-sensing pathway in GBS and provides new insights into the virulence regulation mechanism of GBS. PMID- 17133705 TI - Cancer deaths rates continue to drop. PMID- 17133706 TI - NIH to map changes of 3 cancers. PMID- 17133707 TI - NIH grants $117 million in COBRE awards. PMID- 17133708 TI - Casualty estimates for the invasion of Japan; the "postwar creation" myth. PMID- 17133709 TI - Asian American history: reflections on imperialism, immigration, and "the body". PMID- 17133710 TI - Congress and the welfare state: some historical reflections. PMID- 17133711 TI - [The work of Richard Goldschmidt: an attempt at a synthesis of genetics, developmental biology, and evolutionary theory surrounding the concept of homeosis]. PMID- 17133712 TI - The Miss America Pageant: pluralism, femininity, and Cinderella all in one. PMID- 17133713 TI - Regulation of alkaloid biosynthesis in plants. PMID- 17133714 TI - Ergot alkaloids--biology and molecular biology. AB - EA have been a major benefit, and a major detriment, to humans since early in recorded history. Their medicinal properties have been used, and continue to be used, to aid in childbirth, with new uses being found in the treatment of neurological and cardiovascular disorders. The surprisingly broad range of pharmaceutical uses for EA stems from their affinities for multiple receptors for three distinct neurotransmitters (serotonin, dopamine, and adrenaline), from the great structural diversity of natural EA, and from the application of chemical techniques that further expand that structural diversity. The dangers posed by EA to humans and their livestock stem from the ubiquity of ergot fungi (Claviceps species) as parasites of cereals, and of related grass endophytes (Epichloe, Neotyphodium, and Balansia species) that may inhabit pasture grasses and produce toxic levels of EA. Further concerns stem from saprophytic EA producers in the genera Aspergillus and Penicillium, especially A. fumigatus, an opportunistic pathogen of humans. Numerous fungal species produce EA with a wide variety of structures and properties. These alkaloids are associated with plants in the families Poaceae, Cyperaceae, and Convolvulaceae, apparently because these plants can have symbiotic fungi that produce EA. Pharmacological activities of EA relate to their specific structures. Known as potent vasoconstrictors, the ergopeptines include a lysergic acid substituent with an amide linkage to a complex cyclol lactam ring structure generated from three amino acids. Simpler lysergyl amides and clavines are more apt to have oxytonic or psychotropic activities. One of the lysergyl amides is LSD (5), the most potent hallucinogen known. The EA biosynthetic pathway in Claviceps species has been studied extensively for many decades, and recent studies have also employed epichloes and A. fumigatus. The early pathway, shared among these fungi, begins with the action of an aromatic prenyl transferase, DMATrp synthase, which links a dimethylallyl chain to L tryptophan. When the dmaW gene encoding DMATrp synthase was cloned and sequenced, the predicted product bore no identifiable resemblance to other known prenyl transferases. The dma W genes of Claviceps species are present in clusters of genes, several of which also have demonstrated roles in EA biosynthesis. In many other fungi, dma W homologues are identifiable in otherwise very different gene clusters. The roles of DMA Trp synthase homologues in these other fungi are probably quite variable. One of them is thought to prenylate the phenolic oxygen of L-tyrosine, and another catalyzes the unusual reverse prenylation reaction in the biosynthesis of fumigaclavine C(10), an EA characteristic of A. fumigatus. The second step of the EA pathway is N-methylation of DMATrp (12) to form 13, which is then subjected to a series of oxidation/oxygenation and reduction reactions to generate, in order, chanoclavine-I (16), agroclavine (19), and elymoclavine (6). Shunt reactions generate a wide variety of other clavines. Two epimerizations occur in this pathway: one from 12 to 16, the other from 16 to 19. Further oxidation of 6, catalyzed by the cytochrome-P450 CloA, generates lysergic acid (1). An unusual NRPS complex, lysergyl peptide synthetase (LPS), is responsible for linking 1 to three hydrophobic L-amino acids to generate the ergopeptide lactams. The LPS complex includes two polypeptides, one (LPS 2) possessing a single module for activation of 1, and the other (LPS 1) possessing three modules, each specifying one of the L-amino acids. Variations in LPS 1 sequences are associated with variations in the incorporated amino acids, leading to differences between strain chemotypes, and even multiple ergopeptines within strains. For example, C. purpurea P1 produces two distinct ergopeptines (ergotamine (4) and ergocryptine (Table I)), each of which is believed to be generated by multiple LPS 1 subunits encoded by separate, but related, genes (lpsA1 and lpsA2). The main ecological roles of EA in nature are probably to protect the fungi from consumption by vertebrate and invertebrate animals. The EA produced by plant-symbiotic fungi (such as epichloe endophytes) may protect the fungus by protecting the health and productivity of the host, which may otherwise suffer excessive grazing by animals. The EA, at levels typical of plants bearing these symbionts, can negatively affect the health of large mammals as well herbivorous insects. Some clavines have substantial anti-bacterial properties, which might protect the fungus and, in some cases, their host plants from infection. However, the fact that a large number of epichloe, and even several Claviceps species, produce no detectable EA indicates that the selection for their production is not universal. An unfortunate fact for many livestock producers is that some of the most popular forage grasses tend to possess EA producing epichloe endophytes. Such endophytes are easily eliminated, but confer such fitness enhancements to their hosts that their presence is often preferred, despite the toxic EA. The future looks promising for continued interest in EA. Research continues into their pharmacological properties, medicinal uses, and structure-function relationships. New clavines and lysergic acid derivatives are identified regularly from new sources, such as marine animals. Also, programs are well underway to modify or replace epichloe endophytes of forage grasses in order to produce new grass cultivars that lack these toxins. PMID- 17133715 TI - Chemical and biological aspects of Narcissus alkaloids. PMID- 17133716 TI - Bisindole alkaloids. PMID- 17133717 TI - [Remote effects at cell and subcell level in the hemopoietic system after chronic radiation exposure in man]. AB - The parameters characterizing the state of hemopoietic cells obtained from chronically exposed residents of the Techa riverside villages studied at late time after the exposure included: the level of somatic mutations in the TCR gene, the level of chromosome aberrations, the intensity of peripheral blood lymphocyte apoptosis. Exposed versus unexposed subjects (controls) showed an increased frequency of CD3-CD4+ T-lymphocytes, chromosome aberrations of stable type (translocations) and unstable type (dicentrics, rings), and also increased intensity of lymphocyte apoptosis. The findings of tests using a standard additional gamma-irradiation (1 Gy) accompanied by 24-hour incubation indicated that the rate of apoptosis of lymphocytes was significantly higher in exposed individuals in comparison with unexposed ones. It was suggested basing on the obtained data that at late time the chronic (for over 50 years) exposure at RBM doses from 0.01 to 3.22 Sv was a factor inducing the damage to the genetic apparatus of hemopoietic cells. Evidently, the initial chronic low-intensity irradiation in the above-indicated dose range activates adaptive processes at the cellular level in hemopoietic cells. Late time after the onset of exposure the adaptation reserves are depleted in chronically exposed persons which brings about its failure in the case of a challenge by additional external exposures. PMID- 17133718 TI - [Effect of low-level irradiation on incidence rate and development of malignant neoplasms]. AB - Low-level irradiation (1.2-2.4 cGy, dose-rate 0.6 cGy/day) leads to a significant acceleration of the development of spontaneous leukosis in AKR mice: decrease in the average and the maximum life-span of animals leukosis-carriers (by 20 and 120 days, respectively) and an increase in the leukosis incidence rate (%). The introduction of antioxidant phenozan (beta-(4-hydroxy-3,5-ditertbutylphenyl) propionic acid) results in a considerable antileukosis effect: the average life span increases by more than 40 days and the leukosis incidence rate decreases by 6%. PMID- 17133719 TI - [Migration characteristics and forms of 90Sr and 137Cs observed in the bottom sediments of certain lacustrine ecosystems on the East Urals Radioactive Trace]. AB - The research findings summarized in the paper include data on the radioecological conditions in two lakes (Large Ighish and Small Ighish) located on the East Urals Radioactive Trace. The focus of the study was on the chemical composition of water and bottom sediments, as well as on the specific features of 90Sr and of 137Cs distribution and the forms in which they occur in silts. Coefficients of radionuclide accumulation in the bottom deposits of the lakes of interest were estimated. A conclusion was drawn about the dependences influencing radionuclide distribution by layers of the lake bottom sediments. PMID- 17133720 TI - [Radiation and mutation induction in the human germline]. AB - This review describes the results of some publications on monitoring radiation induced mutation in the human germline. Recent data have shown that minisatellite loci provide a useful and sensitive experimental approach for monitoring radiation-induced mutation in humans. The progress made in validating this approach and the results of recent publications on the analysis of minisatellite mutation rates in the irradiated families are presented. PMID- 17133721 TI - [Genetic aspects of the irradiation in small doses of laboratory lines and experimental populations Drosophila melanogaster]. AB - One of explanations of revealed effects of small doses of an irradiation is induced genetic instability on which background there is a realization of the various radiobiological reactions resulting as to stimulation, and significant oppression of the vital functions of a cell or an organism. In work the given estimations of consequences of an irradiation in small doses of mutant lines of the drosophila are submitted. Paramount value in definition of their size and an orientation of reaction of a genotype is supposed, that, have processes leaders to change of activity of mobile genetic elements and programmed destruction of a cell. PMID- 17133722 TI - [Multidrug resistance determinants in acute myeloid leukemia developed in persons exposed to ionizing radiation due to the Chernobyl accident]. AB - The results of multidrug resistance determinants expression analysis on leukemic cells of 56 acute myeloid leukemia (AML) patients by immunophenotyping are presented. Of these, there were 21 persons exposed to ionizing radiation due to the Chemobyl accident with radiation-associated AML and 35 patients with spontaneous leukemia. The aim of this study was to determine if transport proteins (P-glycoprotein, LRP, and MDR1), apoptosis-related proteins (Fas, Bcl-2, Bax, p53, and Bcl-X(L)), and topoisomerase IIalpha expression in AML patients with the history of radiation exposure differed from those in spontaneous AML cases. Leukemic cells in patients with radiation-associated diseases compared to spontaneous AML more often overexpressed antiapoptotic oncoprotein Bcl-2 (12/21 vs. 6/35, p < 0.005) and less often demonstrated expression of Fas receptor (12/21 vs. 30/35, p < 0.05). Moreover, leukemic cells were simultaneously Fas negative and Bcl-2 positive in 4 out of 21 patients exposed to ionizing radiation but none of spontaneous cases had similar phenotype (p < 0.05). Leukemic cells in patients with radiation-associated AML compared to spontaneous cases more often were P-glycoprotein positive (12/20 vs 9/31, p < 0.05). P-glycoprotein overexpression significantly correlated with resistant disease in patients with radiation-associated AML (r = 0.47, p < 0.05), but was not a prognostic variable for the treatment outcome in terms of overall survival. Defects in pathways of drug-induced apoptosis and function of pump, that actively effluxes drugs could contribute significantly to developing drug resistance in radiation-associated AML. PMID- 17133723 TI - [The absence of the facts connected with the genomic instability after the irradiation in low doses by radiation with low LET]. AB - In the review which is a brief account of more complete document (Koterov A.N. // Int. J. Low Radiat. 2005. V. 1. No. 4. P. 376-451) the data of world researches devoted to a phenomenon of radiation-induced genomic instability (RIGI) are considered. The purpose of the review is the definition of the bottom limit of radiation doses which induced of RIGI in experiments at different methodical approaches (irradiation in vitro, in vivo, in utero, bystander effect and transgeneration effects of radiation). The action only radiation with low LET is examined. Among several hundreds works wasn't revealed any fact, when RIGI induced by low doses irradiation (up to 0.2 Gy) for normal cells and for organism left from maternal womb. Six exceptions are revealed which are named as "apparent" so in all cases the abnormal, unstable, defective objects or ambiguous final parameter were used. Thus, RIGI at low doses of radiation with low LET is a myth. PMID- 17133724 TI - [90Sr and 137Cs in higher aquatic plants of some water basins on the East-Urals Radioactive Trace: species features of radionuclide concentration]. AB - The paper summarizes the results from radioecological studies of a number of water basins located on the East-Urals Radioactive Trace (EURT), as exemplified by the lakes Large Ighish, Small Ighish and Shablish which are included in the medium-distance and remote zone of the EURT. The lake Misyash which is situated in the opposite direction from the release vector and has been only contaminated due to global fallouts has been used as a control water basin. The current species composition of the high aquatic plants and species-specific concentration of 90Sr and 137Cs in them was studied, also, the radionuclide accumulation and discrimination coefficients were estimated. PMID- 17133725 TI - [The influence of chronic and of acute irradiation on the polymorphism of RAPD markers in offspring for irradiated mice]. AB - On mice lines BALB/c and CBA/lac was performed the study of molecular-genetics effects in mice progeny after the chronic (dose rate -0.0017 Gy/day, total dose 0.36 Gy) and acute (dose range 1-3 Gy) exposure of y-radiation on the parents. For variability analysis was used technique of amplification DNA with series of random primers (RAPD-assay). Random primers were used as single primer and in mixture of ones. In this work were held the comparative analysis of the genetic radiosensitivity for stem spermatogonia and spermatides. After the acute exposure the dose dependence for levels of polymorphism of RAPD-markers were obtained. After the chronic irradiation, significant differences from control group were obtained only by use primers mixture M1. Comparative analysis of the genetic radiosensitivity of different stages of mice spermatogenesis are display is similar sensitivity of stem spermatogonia and spermatides after doses of irradiation 1 Gy and 3 Gy. Indicated that after irradiation by dose 2 Gy, spermatogonia are more sensitivity than spermatides. PMID- 17133726 TI - [Radioecological research of the Irtysh-River's and Ob-River's water, base and flood depositions]. AB - The radioecological research of Irtysh-river and Ob-river was held. The content of 137Cs in Irtysh water was compounded 0.62-1.23 Bq/m3, in Ob-- 0.24-0.27 Bq/m3, and the one of 90Sr in Irtysh-- 10-20 Bq/m3, and in Ob-- 5-10 Bq/m3, that is much lower than the permissible sanitary-hygienic norms for the population. The 137Cs stores density on Irtysh-river input lease was compounded 2.7 kBq/m2, is almost in 11 times slashed downstream and is peer 245 kBq/m2 before the Irtysh-river lockin. The 90Sr stores density also was slashed in surveyed leases with 212 down to 106 Bq/m2. Two variants of integrated stores of 137Cs and of 90Sr in flood of the Irtysh-river was held. The balance calculation of annual radionuclides sinks confirms the dominant amount of 137Cs and of 90Sr in downstream Ob-river leases acts now on the Ob's sleeve, instead of from the Irtysh-river as it was supposed earlier. The 137Cs medial annual inflow from the Ob's sleeve almost is in 2 times, and the 90Sr inflow is in 2.3 times more, than are acts from Irtysh-river sleeve. PMID- 17133727 TI - [Compensation mechanisms in hemopoietic stem cell pool (CFUs) under the conditions of experimental chronic gamma-irradiation]. AB - The kinetics, proliferation and differentiation potentials of hemopoietic stem cells (CFUs) of bone marrow and spleen were investigated in CBA-line mice in the early period (1-30 days) of chronic gamma-irradiation at a dose rate of 0.16 Gy/day to attain a cumulative dose of 4.8 Gy. The results of the experimental study showed the prevalent maintenance of productivity of granulocytic and erythrocytic hemopoietic cell series within the range of reference values, persistent inhibition of the megakaryocytic series (in terms of all hemopoiesis parameters of interest), more marked suppression of the population of polypotential CFUs in the bone marrow as compared with that in the spleen. The obtained results indicated that the mechanisms of hemopoiesis compensation at stem cell pool level were as follows: the increase in proliferation potency of erythrocytic and in polypotential precursors, the rise in the proportion of granulocytic precursors in the real differentiation potential of CFUs, and the processes of repopulation manifested with different intensity in all stem cell populations under study. For maintenance of the necessary productivity of CFUs in each of hemopoietic cell series, consecutively or simultaneously, several compensatory-adaptive mechanisms are started, which allows the avoidance of a sharp competition between hemopoietic cell series under the conditions of stem cell pool depopulation, and preservation of the hemopoiesis as a whole. PMID- 17133728 TI - [The grounds of interspices difference in radiation-genetic effects of low-level radiation]. AB - Radiation-genetic effects are characterized by large quantitative difference. After the low dose radiation appearance of mutation is the largest in Drosophila, substantially lower in mice, especially if irradiation was protracted, and practically absent in humans. As the possible grounds of this difference the author consider the existence of non-specific repair mechanism, which has arouse during evolution for diminution of different detrimental impacts, including ionizing radiation. The efficacy of such mechanism of natural protection is determined by the efficacy of the repair mechanisms, which is elevated in compliance with specific life span, which is of necessity for its maintenance. The evolution has led to maximal development of this mechanism in human beings, and it is proved in particular in prevention of the hereditary effect of low level radiation. PMID- 17133729 TI - Esophagus cancer and radiation exposure due to nuclear test fallout: an analysis based on the data of the Semipalatinsk historical cohort, 1960-1999. AB - This paper describes the Semipalatinsk historical cohort study and, in particular, examines the association between combined external and internal radiation exposure and esophagus cancer. Esophagus cancer is the most frequent single cancer site in the cause of death follow-up for the Semipalatinsk cohort. Set up in the 1960s, this historical cohort included 10 exposed settlements in the vicinity of the Semipalatinsk nuclear test site in East Kazakhstan as well as 6 comparison settlements in a low exposure area of the same region. The external and internal radiation doses to the population of the settlements under study were mainly due to local fallout from atmospheric nuclear testing (1949-1962). The database includes dosimetry and health information for 19.545 inhabitants of exposed and comparison villages in the Semipalatinsk region, comprising a total of 582.750 person-years of follow-up between 1960 and 1999. Cumulative effective dose estimates in this cohort range from 20 mSv to -4 Sv, with a mean dose of 634 mSv in the exposed group. Relative risks were calculated in terms of rate ratios, using a Poisson regression model for grouped person-time data. Esophagus cancer was found substantially elevated, with a statistically significant increase of the relative risk with dose and an ERR/Sv of 2.37 (1.45; 3.28) for the total cohort. If the data set was restricted to the exposed group only, the ERR/Sv was found considerably lower (0.18 (-0.16; 0.52)), whereas the dose-response remained significant only in women. Overall, our results based on the Semipalatinsk historical cohort indicate an association between fallout exposure and the risk of esophagus cancer that should be further investigated. PMID- 17133730 TI - A critical survey of fish measurements in populations of the southern Urals. AB - A critical survey of all published measurements made so far aimed at retrospective biological dosimetry using fluorescence in situ hybridisation (FISH) techniques on some workers at the Mayak reprocessing plant and on members of the Techa River cohort is given. Each individual has a recorded dose derived from personal monitoring measurements, usually external gamma-rays for Mayak workers or from reconstruction techniques, usually internally derived for the Techa River cohort. From the person's age, which affects the control level, and the stated dose, an expected number of translocations is calculated for each individual and comparisons made to the observed numbers of translocations. From this, an assessment of how well FISH studies can help to validate existing estimates of dose is made. This varies from study to study. Good agreement is generally obtained for the Techa River cohort and lower doses of the Mayak cohort. Rather poorer agreement applies to the more highly exposed Mayak workers. Some of the discrepancy could be because the FISH painting technique was new and was applied to populations before a proper investigation on how to use it for retrospective biological dosimetry had taken place. In addition, too few cells were generally scored per individual so that statistical uncertainties were large. PMID- 17133731 TI - The application and adaptation of ICRP internal dosimetry models to the calculation of bone marrow tissue doses from 90Sr for epidemiological studies of Techa River populations. AB - The operation of the Mayak Production Association in the Southern Urals region of Russia, resulted in releases of large amounts of radioactive effluent into the Techa River during the period 1949-1956. The residents of the riverside communities were thus exposed to both external radiation, and internal radiation following ingestion of contaminated water and foodstuffs. One of the most important radionuclides for internal exposure was 90Sr. This paper gives a brief overview of the models provided by International Commission on Radiological Protection (ICRP), which are of interest for assessing internal doses from 90Sr. The application of these models to the calculation of red bone marrow doses for the fetus and infant from 90Sr intakes by the mother and the infant is illustrated by an example. A hypothetical individual born in 1951 is used as an example for dose calculations. The following doses due to intakes of 90Sr are taken into account: received in utero due to maternal intakes during pregnancy; received after birth from 90Sr accumulated by the fetus in utero; from intakes in breast milk; from intakes in the infant's diet after weaning. It is shown that doses to the fetus following maternal ingestion and subsequent transfer to the fetus via the placenta dominate the doses received for this particular individual for the first two years of life. Doses to the infant from intakes in breast milk are substantially lower but do make significant contributions to total doses in the first two years after birth. By about the age of two years residual 90Sr from placental transfer still contributes about the same dose as do intakes by the infant, but in later years doses from intakes by the infant dominate. PMID- 17133732 TI - Hypolactasia as a molecular basis of lactose intolerance. AB - Lactase-phlorizin hydrolase (LPH), a membrane-bound glycoprotein present in the luminal surface of enterocytes in the intestine is responsible for lactose intolerance, a phenomenon prevalent in humans worldwide. In the rodent intestine, the post-natal development of the LPH follows a specific pattern, such that the enzyme levels are high in the peri-natal period, but declines considerably upon maturation. The observed maturational decline in the LPH activity is very similar to adult-type hypolactasia observed in humans. Majority of the studies have been carried out using animal models or cell lines and a number of hypotheses have been put forward to explain the maturational decline of lactase activity such as: (a) decreased amount of lactase protein, (b) defect in post-translational modification of precursor lactase to the mature enzyme, and (c) synthesis of an inactive, high molecular weight lactase with altered glycosylation, however, the precise underlying mechanism of adult-type hypolactasia remains undefined. The present review describes the recent developments in understanding the regulation of lactase expression and the possible mechanism of adult-type hypolactasia, as a cause of lactose intolerance. PMID- 17133733 TI - Biochemistry of homocysteine in health and diseases. AB - The amino acid homocysteine (Hcy), formed from methionine has profound importance in health and diseases. In normal circumstances, it is converted to cysteine and partly remethylated to methionine with the help of vit B12 and folate. However, when normal metabolism is disturbed, due to deficiency of cystathionine-beta synthase, which requires vit B6 for activation, Hcy is accumulated in the blood with an increase of methionine, resulting into mental retardation (homocystinuria type I). A decrease of cysteine may cause eye diseases, due to decrease in the synthesis of glutathione (antioxidant). In homocystinurias type II, III and IV, there is accumulation of Hcy, but a decrease of methionine, thus, there is no mental retardation. Homocysteinemia is found in Marfan syndrome, some cases of type I diabetes and is also linked to smoking and has genetic basis too. In hyperhomocysteinemias (HHcys), clinical manifestations are mental retardation and seizures (type I only), ectopia lentis, secondary glaucoma, optic atrophy, retinal detachment, skeletal abnormalities, osteoporosis, vascular changes, neurological dysfunction and psychiatric symptoms. Thrombotic and cardiovascular diseases may also be encountered. The harmful effects of homocysteinemias are due to (i) production of oxidants (reactive oxygen species) generated during oxidation of Hcy to homocystine and disulphides in the blood. These could oxidize membrane lipids and proteins. (ii) Hcy can react with proteins with their thiols and form disulphides (thiolation), (iii) it can also be converted to highly reactive thiolactone which could react with the proteins forming -NH-CO- adducts, thus affecting the body proteins and enzymes. Homocystinuria type I is very rare (1 in 12 lakhs only) and is treated with supplementation of vit B6 and cystine. Others are more common and are treated with folate, vit B12 and in selected cases as in methionine synthase deficiency, methionine, avoiding excess. In this review, the role of elevated Hcy levels in cardiovascular, ocular, neurologial and other diseases and the possible therapeutic measures, in addition to the molecular mechanisms involved in deleterious manifestations of homocysteinemia, have been discussed. PMID- 17133734 TI - Characterization of carotenoid hydroxylase gene promoter in Haematococcus pluvialis. AB - Astaxanthin, a high-value ketocarotenoid is mainly used in fish aquaculture. It also has potential in human health due to its higher antioxidant capacity than beta-carotene and vitamin E. The unicellular green alga Haematococcus pluvialis is known to accumulate astaxanthin in response to environmental stresses, such as high light intensity and salt stress. Carotenoid hydroxylase plays a key role in astaxanthin biosynthesis in H. pluvialis. In this paper, we report the characterization of a promoter-like region (-378 to -22 bp) of carotenoid hydroxylase gene by cloning, sequence analysis and functional verification of its 919 bp 5'-flanking region in H. pluvialis. The 5'-flanking region was characterized using micro-particle bombardment method and transient expression of LacZ reporter gene. Results of sequence analysis showed that the 5'-flanking region might have putative cis-acting elements, such as ABA (abscisic acid) responsive element (ABRE), C-repeat/dehydration responsive element (C repeat/DRE), ethylene-responsive element (ERE), heat-shock element (HSE), wound responsive element (WUN-motif), gibberellin-responsive element (P-box), MYB binding site (MBS) etc., except for typical TATA and CCAAT boxes. Results of 5' deletions construct and beta-galactosidase assays revealed that a highest promoter-like region might exist from -378 to -22 bp and some negative regulatory elements might lie in the region from -919 to -378 bp. Results of site-directed mutagenesis of a putative C-repeat/DRE and an ABRE-like motif in the promoter like region (-378 to -22 bp) indicated that the putative C-repeat/DRE and ABRE like motif might be important for expression of carotenoid hydroxylase gene. PMID- 17133735 TI - Purification and characterization of trehalose-6-phosphate synthase from Saccharomycopsis fibuligera A11. AB - Mutant A11, a mutant of Saccharomycopsis fibuligera Sdu with low acid and neutral trehalase was found to accumulate over 18% (w/w) trehalose from starch in its cells. In this study, trehalose-6-phosphate synthase (Tps1) was purified to homogeneity from this mutant, with a 30-fold increase in the specific enzyme activity, as compared to the concentrated cell-free extract, from initial cells. The molecular mass of the purified enzyme as determined by SDS-PAGE was 66 kDa. The optimum pH and temperature of the purified enzyme were 6.6 and 37 degrees C, respectively. The enzyme was activated by Ca2+, K+ and Mg2+, with K+ showing the highest activation at 35 mM. On the other hand, Mn2+, Cu2+, Fe3+, Hg2+ and Co2+ inhibited the enzyme. The enzyme was also strongly inhibited by protease inhibitors such as iodoacetic acid, EDTA and PMSF. PMID- 17133736 TI - Isolation and identification of alpha-amylase producing Bacillus sp. from dhal industry waste. AB - A bacterial strain was isolated from dhal industry red gram waste and identified as Bacillus. A thermostable extracellular amylase was partially purified from the strain. Optimum temperature and pH for the enzyme were found to be 60 degrees C and 6.5, respectively. The maximum amylase production was achieved with maltose as carbon source. Among the nitrogen sources, peptone and yeast extract produced maximum amylase. PMID- 17133737 TI - Hepatoprotective effect of aqueous extract of Phyllanthus niruri on nimesulide induced oxidative stress in vivo. AB - Nimesulide (NIM), an atypical non-steroidal anti-inflammatory drug (NSAID) is also used as analgesic. In the present study, we evaluated its effect on the prooxidant-antioxidant system of liver and the hepatoprotective potential of aqueous extract of the herb Phyllanthus niruri (PN) on NIM-induced oxidative stress in vivo using a murine model, by determining the activities of hepatic anti-oxidant enzymes superoxide dismutase (SOD) and catalase (CAT), levels of reduced glutathione (GSH) and lipid peroxidation (expressed as malonaldialdehyde, MDA). Aqueous extract of PN at a dose of 50 or 100 mg/kg body wt was administered either intraperitoneally or orally for 7 days, before NIM administration at a dose of 8 mg/kg body wt twice daily for 7 days in mice. Animals were sacrificed 24 h after administration of final dose of NIM. In another set of experiments, both aqueous extract of PN (at a dose of 50 or 100 mg/kg body wt) and NIM (8 mg/kg body wt) were administered simultaneously for 7 days. Animals were sacrificed 24 h after administration of final dose of the extract and NIM, liver tissues were collected, and the activities of SOD and CAT and levels of GSH and lipid peroxidation end-product (as MDA), were determined from the livers of all the experimental animals. Appropriate NIM control was maintained for all sets of experiments. NIM administration (8 mg/kg body wt) for 7 days caused significant depletion of the levels of SOD, CAT and reduced GSH, along with the increased levels of lipid peroxidation. Intraperitoneal administration of the extract at a dose of 50 mg/kg body wt for 7 days,. prior to NIM treatment, significantly restored most of the NIM-induced changes and the effect was comparable to that obtained by administering 100 mg/kg body wt of the extract orally. Thus, results suggested that intraperitoneal administration of the extract could protect liver from NIM-induced hepatic damage more effectively than oral administration. Antioxidant property of the aqueous extract of PN was also compared with that of a known potent antioxidant, vitamin E. The PN extract at a dose of 100 mg/kg body wt along with NIM was more effective in suppressing the oxidative damage than the PN extract at a dose of 50 mg/kg body wt. Results suggested that beneficial effect of the aqueous extract of PN, probably through its antioxidant property, might control the NIM-induced oxidative stress in the liver. PMID- 17133738 TI - Protective effects of silymarin, a milk thistle (Silybium marianum) derivative on ethanol-induced oxidative stress in liver. AB - The production of reactive oxygen species (ROS) is considered to be a major factor in oxidative cell injury. The antioxidant activity or the inhibition of the generation of free radicals is important in providing protection against such hepatic damage. Silymarin, derived from the milk thistle plant, Silybium marianum, has been used in traditional medicine as a remedy for diseases of the liver and biliary tract. In the present study, the effect of hepatoprotective drug silymarin on body weight and biochemical parameters, particularly, antioxidant status of ethanol-exposed rats was studied and its efficacy was compared with the potent antioxidant, ascorbic acid as well as capacity of hepatic regeneration during abstention. Ethanol, at a dose of 1.6 g/kg body wt/day for 4 wks affected body weight in 16-18 week-old male albino rats (Wistar strain weighing 200-220 g). Thiobarbituric acid reactive substance (TBARS) level, superoxide dismutase (SOD), and glutathione-s-transferase (GST) activities were significantly increased, whereas GSH content, and catalase, glutathione reductase (GR) and GPx (glutathione peroxidase) activities significantly reduced, on ethanol exposure. These changes were reversed by silybin and ascorbic acid treatment. It was also observed that abstinence from ethanol might help in hepatic regeneration. Silybin showed a significant hepatoprotective activity, but activity was less than that of ascorbic acid. Furthermore, preventive measures were more effective than curative treatment. PMID- 17133739 TI - Photoilluminated riboflavin/riboflavin-Cu(II) inactivates trypsin: Cu(II) tilts the balance. AB - Riboflavin (RF) upon irradiation with fluorescent light generates reactive oxygen species like superoxide anion, singlet and triplet oxygen, flavin radicals and substantial amounts of hydrogen peroxide (H2O2). H2O2 can freely penetrate cell membrane and react with a transition metal ion like Cu(ll), generating hydroxyl radical via the modified metal-catalyzed Haber-Weiss reaction. Earlier, it was reported that trypsin-chymotrypsin mixture served as an indirect antioxidant and decreased free radical generation. Thus, in the present study, we used photoilluminated RF as a source of ROS to investigate the effect of free radicals on the activity of trypsin. We also compared the damaging effect of photoilluminated RF and RF-Cu(ll) system using trypsin as a target molecule. RF caused fragmentation of trypsin and the effect was further enhanced, when Cu(II) was added to the reaction. Results obtained with various ROS scavengers suggested that superoxide radical, singlet and triplet oxygen were predominantly responsible for trypsin damage caused by photoilluminated RF. On the other hand, when Cu(ll) was added to the reaction, hydroxyl radical was mainly responsible for trypsin damage. A mechanism of generation of various ROS in the reaction is also proposed. Trypsin did not show any antioxidant effect with RF alone or with RF-Cu(II) combination. PMID- 17133740 TI - Identification of alpha-2u globulin in the rat preputial gland by MALDI-TOF analysis. AB - The low molecular mass proteins found in the pheromonal sources such as urine, saliva, glandular secretion etc have been reported as ligand carriers for the processes of chemocommunication in mammals. The preputial gland plays an important role in the production of olfactory signals for pheromonal communication. Thus, in the present study, alpha-2u globulin having molecular mass of 18 kDa has been identified in the preputial gland of Norway rat (Rattus norvegicus) by in-gel trypsin digestion and analyzing the resulting peptides by MALDI-TOF. Since preputial gland is one of the major pheromonal sources in rat, the results suggest that alpha-2u globulin might act as a carrier for hydrophobic odorants of preputial gland. PMID- 17133741 TI - Extraction and carrier-facilitated transport of amino acids using synthetic non cyclic receptors through bulk liquid membrane. AB - The extraction and carrier-facilitated transport of amino acids (leucine, valine and glycine) was studied through chloroform bulk liquid membrane system using a series of non-cyclic receptors such as diethylene glycol (1), diethylene glycol dimethyl ether (2), diethylene glycol dibutyl ether (3), diethylene glycol dibenzoate (4), triethylene glycol (5) and tetraethylene glycol (6). The amount of amino acid extracted and transported depends mainly upon the structure and the concentration of the receptors and also on the concentration of amino acid. The receptors 1 to 4, having small chain length and flexible end groups, formed stable complexes with amino acids, and the flexibility of receptors in different conformational forms was responsible for their carrier ability, while the receptors 5 and 6, having larger chain length showed poor carrier ability. Hydrophobicity of amino acids also play an important role in the extraction as well as transport process. PMID- 17133742 TI - I'm not one to keep secrets: how I'll vote this fall. PMID- 17133743 TI - Will we ever take any personal responsibility? PMID- 17133744 TI - "Animal rights" terrorists/extremists: A serious threat to medicine. PMID- 17133745 TI - Pathological gambling in Missouri. AB - Riverboat casinos were introduced in 1994, and there are now eleven gambling venues in Missouri, in addition to the lottery. Gambling is monitored by the Missouri Gaming Commission which was established to supervise gambling operations in the state, and minimize criminal involvement. The Commission also operates programs for problem gamblers that are described. Pathological gambling has become a major problem in Missouri and elsewhere, and its characteristics and clinical management are reviewed herein. PMID- 17133746 TI - How to prepare for the medical malpractice trial. PMID- 17133747 TI - Kansas City blues: fighting the insurance cabal in court. PMID- 17133748 TI - DIY--The physician pilot: parallels between aviation and medicine. PMID- 17133749 TI - The role of international medical graduates in American medicine. PMID- 17133750 TI - Is stem cell research the next gambling boat? PMID- 17133751 TI - Tired and sleepy: update on sleep disorders. PMID- 17133752 TI - Snoring and its management. AB - There are numerous strategies, devices and procedures available to treat snoring. The surgical procedures have an overall success rate of 60-70%, but this probably decreases over time, especially if there is weight gain. There are no long-term rigorously-designed studies comparing the various procedures for decreasing snoring. PMID- 17133753 TI - Obstructive sleep apnea: A practical review. AB - Obstructive Sleep Apnea (OSA) is a condition of repetitive stoppage of breathing during sleep. OSA has assumed major clinical importance as it is linked with the pathophysiology of many serious medical conditions such as congestive heart failure and stroke. We shall review the history, epidemiology, clinical presentation, evaluation, and management of sleep apnea. PMID- 17133754 TI - Obstructive sleep apnea: A modifiable risk factor for stroke? AB - Stroke is well-established to be a leading cause of not only death, but also disability. As with any other medical condition, prevention and risk factor modification are preferable to treatment of emergent disease. This underscores the importance of understanding the relationships of obstructive sleep apnea (OSA) to stroke and its risk factors. While the data are conflicting regarding the absolute causative relationship of OSA to stroke, there is a wealth of information relating OSA to many other vascular conditions and risk factors. Given that OSA is relatively easy to diagnose and treat, and the benefits of risk factor modification and improved quality of life are inarguable, appropriate diagnosis and management of OSA become vital. PMID- 17133755 TI - The restless legs syndrome: diagnosis and treatment. AB - Restless leg syndrome (RLS) is a common neurological disorder which can be accompanied with PLMS and can result in sleep disturbance. Greater understanding of pathophysiology of RLS in recent years has led to increased treatment options for this condition. A comprehensive clinical evaluation and appropriate management using combined nonpharmacologic and pharmacologic strategies can result in significant improvement in treatment outcomes in patients with RLS. PMID- 17133756 TI - Narcolepsy & its management. AB - This paper discusses the diagnosis and treatment of narcolepsy. PMID- 17133757 TI - Sleep and depression. AB - Depression results in severe sleep disturbance. The "depressed brain" takes longer to initiate sleep, shortens the period to dreaming sleep, spends little or no time in "deep sleep" and is subject to hyperactive brain regions during sleep. Antidepressants rectify the sleep disturbance and restore normal sleep architecture and presumably normal brain metabolism during sleep. Early intervention may lead to a decrease in neuronal cell loss due to depression, and better long-term outcomes for mood disorders. PMID- 17133758 TI - Sleep and Parkinson's disease. AB - Movement disorders encompass a wide variety of neurological problems, and each has its own constellation of cognitive, motor and sleep problems. Those problems may be secondary to the disease process, the treatments for the disease or both. One of the most commonly seen of these disorders is Parkinson's disease. This article will discuss the sleep issues commonly associated with that disorder, and will present suggestions for treatment. PMID- 17133759 TI - Detecting microalbuminuria and taking action in the cardiometabolic syndrome and type 2 diabetes mellitus. AB - Microalbuminuria is an early indicator of cardiac and renal vascular endothelial damage in individuals with diabetes mellitus, impaired glucose tolerance, hypertension, and the CardioMetabolic Syndrome. This simple, inexpensive dipstick screening test is not only associated with an increased risk of progressive renal insufficiency but also with an increased risk of cardiovascular disease and events. Regular screening for microalbuminuria will allow for early detection and intervention to prevent renal and vascular complications of these disease states. PMID- 17133760 TI - Rheumatoid arthritis: A review. AB - Rheumatoid arthritis (RA) is a systemic autoimmune disease with primary manifestations in the diarthrodial (movable) joints. Methotrexate in weekly doses is the most widely used disease modifying agent. The recent development of biologic agents designed to shut off disease activity can be of great benefit to individuals who fail methotrexate therapy. New treatments such as tumor necrosis factor suppressants are effective in up to 80% of patients. It is extremely important to diagnose RA in its earliest stages so that patients may benefit from antimetabolites and biologics before permanent joint damage takes place. PMID- 17133761 TI - The long road to patient safety: A status report on Missouri acute-care hospitals. AB - We surveyed Missouri's acute-care hospitals twice to assess the "state of the art" in patient safety systems. We found improvements in all areas studied, but progress is modest and falls short of national recommendations. We urge all Missourians to become more knowledgeable and assertive consumers of hospital care, and we ask them to support Missouri hospitals in their patient safety efforts, including provision of financial resources to put necessary systems in place. PMID- 17133762 TI - Social perceptions and preferences of youth with asthma. AB - Peers are a primary source of psychosocial support in youth. Chronic disease such as asthma can make youth feel different and impinge on their adherence to treatment. We investigated factors that make the asthmatic adolescent feel different from peers, and explore their willingness to belong to a peer social group such as an asthma club. Sixty-six youth (ages 8-18 years) with asthma completed an anonymous questionnaire that included both multiple-choice and open ended questions designed to explore the feelings of the respondents. Almost one third of our sample reported negative feelings regarding their asthma. Nearly 27% reported that their diagnosis made them feel different from their healthy peers, while over 25% admitted feeling uncomfortable using their inhaler in front of their friends. Almost one-half of adolescents felt restricted or excluded from school activities, athletics, or social clubs. While most respondents (93.9%) PMID- 17133763 TI - Hydrogen production by photosynthetic green algae. AB - Oxygenic photosynthetic organisms such as cyanobacteria, green algae and diatoms are capable of absorbing light and storing up to 10-13% of its energy into the H H bond of hydrogen gas. This process, which takes advantage of the photosynthetic apparatus of these organisms to convert sunlight into chemical energy, could conceivably be harnessed for production of significant amounts of energy from a renewable resource, water. The harnessed energy could then be coupled to a fuel cell for electricity generation and recycling of water molecules. In this review, current biochemical understanding of this reaction in green algae, and some of the major challenges facing the development of future commercial algal photobiological systems for H2 production have been discussed. PMID- 17133764 TI - Iterative ACORN as a high throughput tool in structural genomics. AB - High throughput macromolecular structure determination is very essential in structural genomics as the available number of sequence information far exceeds the number of available 3D structures. ACORN, a freely available resource in the CCP4 suite of programs is a comprehensive and efficient program for phasing in the determination of protein structures, when atomic resolution data are available. ACORN with the automatic model-building program ARP/wARP and refinement program REFMAC is a suitable combination for the high throughput structural genomics. ACORN can also be run with secondary structural elements like helices and sheets as inputs with high resolution data. In situations, where ACORN phasing is not sufficient for building the protein model, the fragments (incomplete model/dummy atoms) can again be used as a starting input. Iterative ACORN is proved to work efficiently in the subsequent model building stages in congerin (PDB-ID: lis3) and catalase (PDB-ID: 1gwe) for which models are available. PMID- 17133765 TI - Correlation between biochemical properties and adaptive diversity of skeletal muscle myofibrils and myosin of some air-breathing teleosts. AB - Functional properties of myofibrils and relative stability of myosin of five teleosts Channa punctata, Clarias batrachus, M astacembalus armatus, Labeo rohita and Catla catla adapted to different breathing modes were compared. Myofibrillar contractility and m-ATPase of air-breathing organ (ABO) possessing C.punctata and C. batrachus were low and least affected by pH in the range of 7.1-8.5. However, their myosin isoforms were relatively thermostable, more soluble at sub-neutral pH values, between 0.1 to 0.15 M KCl concentrations and less susceptible to a chymotryptic digestion. In contrast, myofibrils and myosin of water-breather major carps L. rohita and C. catla were more contractile and susceptible to pH and salt concentrations. Thus, correlation between catalytic efficiency and relative stability of myofibrils and myosin of ABO-possessing teleosts was of reverse order and magnitude, as compared to water-breathers. Interestingly, myofibrils and myosin of the behavioral air-breather M. armnatus showed intermediate properties. The specific levels of m-ATPase of all the five teleosts were in conformity with the levels of metabolic marker, the lactate dehydrogenase. The effect of chymotryptic cleavage of 94 and 173 kDa domains on ATPase, individuality of peptide maps of MyHC isomers and perturbation of phenylalanine residues by urea implicated hydrophobic residues in stabilizing myosin structure in these fish. The present study suggests two apparent evolutionary modifications of myofibrils and myosin in ABO-possessing teleosts: (i), 'down-regulation' of ATPase that explains sluggishness of such species and, (ii), more stable molecular structure to support stress of air-breathing modes of life. PMID- 17133766 TI - Physical, X-ray diffraction and scanning electron microscopic studies of uroliths. AB - Identification of chemical constituents of calculus is important in the diagnosis and management of urolithiasis. The compositional variability of uroliths has different etiologies and requires various modes of treatment and prophylaxis. In the present study, we report the chemical compositional analyses of calculi recovered from buck and bullock by X-ray diffraction (XRD) and energy dispersive X-ray (EDX) techniques and ultra-structure examination by scanning electron microscopy (SEM). XRD and EDX investigations conclusively established the chemical compositions of urinary calculi under investigation. The calculus from buck (sample I) had calcium oxalate monohydrate, a dominant salt phase and magnesium compound in significant amount. The calculus from bullock (sample II) had magnesium ammonium phosphate phase, with significant amount of calcium in apatite form and K+ ions. SEM study at higher magnification (X 1000) showed bipyramidal crystals in external zones of urolith (sample I). The struvite apatite calculus showed that basic unit of structure was lamination and the laminitis appeared to be made up of fine granules and high porosity. The bio mineralization process of calculus formation was also studied, with a view to take preventive and therapeutic measures for amelioration of urinary stone diseases in animals and humans. PMID- 17133767 TI - Evaluation of water binding, seed coat permeability and germination characteristics of wheat seeds equilibrated at different relative humidities. AB - The relative binding of seed water and seed coat membrane stability were measured in two contrasting wheat (Triticum aestivum L) varieties, HDR 77 (drought tolerant) and HD 2009 (susceptible) using seed water sorption isotherms, electrical conductivity (EC) of leachates and desorption-absorption isotherms. Analysis of sorption isotherm at 25 degrees C showed that the seeds of HDR 77 had significantly higher number of strong binding sites, with correspondingly greater amount of seed water as strongly bound water, as compared to HD 2009. Total number of binding sites was also higher in HDR 77 than HD 2009, which explained the better desiccation tolerance and higher capacity to bind water in seeds of HDR 77. EC of seed leachate in both varieties did not change with respect to change in equilibrium relative humidity (RII), indicating the general seed coat membrane stability of wheat seeds. However, absolute conductivity values were higher for HD 2009. showing its relatively porous seed coat membrane. Significantly lower area enclosed by the desorption-absorption isotherm loop in HDR 77, as compared to HD 2009 also indicated the greater membrane integrity of HDR 77. Germination and seedling vigour of HD 2009 were reduced when equilibrated over very low and very high RH. In contrast, germination and vigour in HDR 77 were maintained high, except at very high RH, indicating again its desiccation tolerance. Thus, the study demonstrated the relative drought tolerance of HDR 77, on the basis of seed water-binding characteristics and seed membrane stability. Seed membrane stability as measured by seed leachate conductivity or as area under dehydration-rehydration loop may be used as a preliminary screening test for drought tolerance in wheat. PMID- 17133768 TI - Purification of a peroxidase from Solanum melongena fruit juice. AB - Solanum melongena fruit juice contains peroxidase activity of the order of 0.125 IU/mL. A method for the 11-fold purification of the enzyme was developed. The Km values of the peroxidase for the substrates guaiacol and hydrogen peroxide were 6.5 mM and 0.33 mM, respectively. The pH and temperature optima were 5.5 and 84 degrees C, respectively using guaiacol as the substrate. Sodium azide and phenyl hydrazine inhibited the enzyme competitively. PMID- 17133769 TI - Isolation of stress responsive Psb A gene from rice (Oryza sativa l.) using differential display. AB - Differential display (DD) experiments were performed on drought-tolerant rice (Oryza sativa L.) genotype N22 to identify both upregulated and downregulated partial cDNAs with respect to moisture stress. DNA polymorphism was detected between drought-stressed and control leaf tissues on the DD gels. A partial cDNA showing differential expression, with respect to moisture stress was isolated from the gel. Northern blotting analysis was performed using this cDNA as a probe and it was observed that mRNA corresponding to this transcript was accumulated to high level in rice leaves under water deficit stress. At the DNA sequence level, the partial cDNA showed homology with psb A gene encoding for Dl protein. PMID- 17133770 TI - A p53-like protein from a freshwater mollusc Lamellidens corrianus. AB - p53 is the most frequently mutated protein in human cancers and the accumulation of its high levels is a potential novel marker for malignancy. Recently, its homologues such as p63 and p73 have been reported in human, mice and fish. Environmentally induced alterations in p53 protein have been reported to contribute to pathogenesis of leukemia in soft-shell clam Mya arenaria inhabiting polluted water, suggesting that p53 proteins can also be used as pollution markers. In the present study, the presence of p53 protein or its homologues was investigated in tissues of bivalve molluscs Lamellidens corrianus that are predominant in the freshwater riverine environment and are well suited to act as test organisms for evaluation of habitat degradation. The molluscs were collected live from the river Ganga at three sampling sites viz., Kanpur, Allahabad and Varanasi and different tissues (foot, gill and mantle) were collected. Proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE). On immunoblot analysis, a 45 kDa protein (p45) was recognized by the monoclonal anti-p53 antibody in the molluscan tissues. The p45 showed immunoreactivity in all the three tissues of molluscs collected at Kanpur, in foot and gill tissues in those collected at Allahabad, and in foot tissue only, in those collected at Varanasi. Since monoclonal anti-p53 recognizes a denaturation-resistant epitope on the p53 (53 kDa) nuclear protein and does not react with other cellular proteins, the molluscan p45 is a p53-homologue or p53 like protein. Further, the differential expression of p45 in the different organs might serve as a useful biomarker that would help in establishing pollution gradient for environmental monitoring in the large aquatic ecosystems. PMID- 17133771 TI - Nitric oxide levels during erythroid differentiation in K562 cell line. AB - Nitric oxide (NO) is endogenous mediator of numerous physiological processes that range from regulation cardiovascular function and neurotransmission to antipathogenic and tumoricidal responses. This study was designed to investigate the possible role of NO during erythroid differentiation in K562 erythroleukemia cells. The chronic myelogenous leukemia (K562) cell line can be triggered in culture to differentiate along the erythrocytic pathway, in response to a variety of stimulatory agents. In this study, K562 cells were induced to synthesize hemoglobin by hemin. We investigated NOx (nitrate+nitrite) levels in uninduced (control) and hemin-induced K562 cell lysates during erythroid differentiation. Our results showed that NO levels decreased significantly on fourth and sixth day both in hemin-induced and control cells; the decrease was, however, more in hemin induced group than in control group. PMID- 17133772 TI - Preparation of prospective plant oil derived micro-emulsion vehicles for drug delivery. AB - Biocompatible oil-in-water (o/w) micro-emulsions can be prospective drug delivery vehicles for their capability to solubilize lipophilic (oil soluble) drugs in the dispersed oil. Plant oils are considered suitable for such a purpose. In this study, we have attempted to examine the dispersion of corn, cottonseed, clove, orange and peppermint oils, as well as isopropyl myristate (IPM) in water continuum in presence of surfactants Tween-20, Brij-30 and Brij-92 and co surfactants ethanol (EtOH) and isopropyl alcohol (iPrOH). Both ternary (oil/surfactant/water) and psedoternary (oil/surfactant + co-surfactant/water) phase diagrams were constructed. The ternary systems produced larger micro emulsion forming zones than the psedo-ternary systems. The combinations peppermint oil/iPrOH/water, IPM/iPrOH/water and 1:1 (v/v) peppermint oil + IPM/iPrOH/water were found to form fair proportion of single-phase surfactant less micro-emulsion. The surfactant-aided ternary systems produced larger clear microemulsion zones, compared to pseudo-ternary systems, while the behaviour of surfactant-less systems was intermediate. The prepared systems had shelf life of 1 year and they withstood temperature variations in the range of 4-40 degrees C. PMID- 17133773 TI - Peri-operative decreased cAMP levels in long-term alcoholic patients. AB - Long-term alcoholic patients have a five-fold higher risk of post-operative bleeding complications compared with nonalcoholic individuals. Serotonin increases and cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) decrease platelet aggregation. We examined the platelet-rich plasma levels of these substances and agonist-induced platelet aggregation in long-term alcoholic patients before and after surgery. Thirty-three consecutive patients (13 long-term alcoholics and 20 non-alcoholics) scheduled for tumour resections of the upper digestive tract were included in the study. The levels of cAMP were significantly decreased before and after surgery in long-term alcoholic patients, but there were no significant differences in cGMP and serotonin levels in alcoholic compared with non-alcoholic patients. In contrast to previous studies, no significantly altered aggregation responses in long-term alcoholics were found. A possible explanation is decreased inhibition through diminished cAMP levels; cGMP and serotonin do not seem to influence peri-operative haemostasis. PMID- 17133774 TI - Valdecoxib provides effective pain relief following acute ankle sprain. AB - We sought to determine whether valdecoxib is as effective as diclofenac in treating acute ankle sprain. Patients (n=202) with acute first- and second-degree ankle sprain were randomized to valdecoxib (40 mg twice daily on day 1 followed by 40 mg once daily on days 2-7) or diclofenac (75 mg twice daily). The primary efficacy end-point was the Patient's Assessment of Ankle Pain visual analogue scale (VAS, 0-100 mm) value on day 4. Valdecoxib was as efficacious as diclofenac in treating the signs and symptoms of acute ankle sprain. The mean VAS reduction in ankle pain on day 4 was not different between groups; the two-sided 95% confidence interval for the between-group difference was within the prespecified limit for non-inferiority (10 mm). There were no significant differences between groups for all secondary efficacy end-points. The two treatments were similarly effective and well tolerated for treatment of acute ankle sprain. PMID- 17133775 TI - P-wave dispersion is increased in pregnancy due to shortening of minimum duration of P: does this have clinical significance? AB - Most pregnant women complain of palpitation, and various kinds of arrhythmias can be observed during pregnancy. We investigated P-wave and QT dispersion during pregnancy. Healthy pregnant women (n=162) and healthy age-matched, non-pregnant women (n=150) were included. We performed electrocardiography and transthoracic echocardiography and determined serum oestradiol levels in both groups, and performed Holter monitoring in the pregnant group only. Resting heart rate, P wave dispersion, left ventricular diastolic diameter, left atrial diameter and serum oestradiol levels in the pregnant group were significantly higher than in the control group. Minimum P-wave duration was shorter in the control group than in the pregnant group; however, there was no statistically significant difference in maximum P wavelength and corrected QT dispersion between the groups. No atrial fibrillation was detected in the pregnant group during Holter monitoring. Shortening of the minimum P-wave duration leads to increased P-wave dispersion during pregnancy. In contrast to other pathologies with increased P-wave dispersion, paroxysmal atrial fibrillation is absent in pregnant women; this may be a result of the stable maximum P wavelength that is present during pregnancy. PMID- 17133776 TI - Cardioprotective effect of edaravone against ischaemia-reperfusion injury in the rabbit heart before, during and after reperfusion treatment. AB - The free radical scavenger edaravone is able to stimulate prostacyclin release and inhibit the lipoxygenase pathway in the arachidonic acid cascade. The effect of edaravone administration on myocardial damage in rabbit hearts subjected to ischaemia-reperfusion was examined at different times relative to reperfusion. All rabbits underwent sustained coronary artery occlusion for 30 min followed by 3 h of reperfusion. Rabbits were divided into the following groups: control; early (3 mg/kg edaravone IV 10 min before reperfusion); immediate (3 mg/kg edaravone IV immediately after the start of reperfusion); and late (3, 6 or 10 mg/kg edaravone IV 5 min after the start of reperfusion). Single bolus administration of edaravone 10 min before reperfusion or immediately upon initiation of reperfusion appears to be associated with reductions in infarction size and the percentage of apoptotic cells, but treatment with edaravone 5 min after initiation of reperfusion does not appear to have this protective effect. PMID- 17133777 TI - Expression of ephrin in retinal neovascularization and iris rubeosis. AB - We investigated expression of ephrin-B2 and Eph-B4 in the retinal tissues of six primate eyes with neovascularization and iris rubeosis secondary to laser-induced central retinal vein occlusion and in tissue from 10 human eyes with proliferative diabetic retinopathy. Two primate eyes with rubeosis and retinal neovascularization were enucleated 1, 2 and 4 weeks after the creation of central retinal vein occlusion. Antibodies were localized using the avidin-biotin reaction. In the primate eyes, ephrin-B2 was negative at I week and positive at 2 and 4 weeks in the rubeotic tissue, but was positive only at 2 weeks in the retinal neovascular membrane. Eph-B4 was negative in all the primate eye specimens. In the human tissue, ephrin-B2 was detected in two of the five eyes with rubeosis and three of the five eyes with retinal neovascularization. These data suggest that ephrin-B2 is a key regulator of neovascularization. PMID- 17133778 TI - The effect of fexofenadine on pruritus in a mouse model (HR-ADf) of atopic dermatitis. AB - Fexofenadine, a histamine H1-receptor antagonist, is approved for the treatment of pruritus associated with atopic dermatitis. The effects of fexofenadine on scratching behaviour, and plasma levels of histamine and eotaxin were assessed in a new model of atopic dermatitis. Mice fed a diet low in Mg2+ and Zn2+ (special diet S) were compared with mice on a normal diet (N) or diet S plus fexofenadine HCl for weeks 0-10 (S + F(0-10)), 0-5 (S + F(0-5)) or 6 - 10 (S + F(6-10)) (seven mice per group). Compared with group N, group S mice showed significantly greater scratching frequency, and plasma histamine and eotaxin concentrations; these three variables were significantly lower in group S + F(0-10) than in group S. Scratching frequency increased when fexofenadine was discontinued. Fexofenadine significantly reduced mast cell and eosinophil numbers. Histamine may be important in the pathological changes seen in this model of atopic dermatitis, suggesting that it might aid future development of antihistamines for the treatment of atopic dermatitis. PMID- 17133779 TI - Prevention of gynaecological adhesions using haemostatic fleece in a rabbit model. AB - This pre-clinical study was performed to investigate the ability of the haemostatic fleece TachoComb to prevent adhesion formation following uterine surgery. Thirty rabbits were randomized to receive TachoComb or no intervention following incision to the right uterine horn. After 14 days, the animals were killed and examined for the presence of adhesion. The lengths of any adhesions were measured and the severity was recorded as a score (0, no adhesion; 1, adhesion easy to lyse; 2, adhesion lysed with traction; 3, adhesion separated by sharp dissection). The incidence of adhesions was 100% in the control group compared with 33% in the TachoComb-treated animals. The mean adhesion score was significantly lower (0.7 versus 2.2) and the mean adhesion length category was significantly shorter (0.4 versus 2.0) with TachoComb than in the control group. This study indicates that TachoComb is a well-tolerated and effective means of preventing adhesion following gynaecological surgery. PMID- 17133780 TI - Nourkrin: objective and subjective effects and tolerability in persons with hair loss. AB - This randomized, double-blind, placebo-controlled study was designed to investigate the efficacy and tolerability of Nourkrin, a new natural agent for the treatment of hair loss based on marine proteins, and minerals and vitamins. Fifty-five subjects with hair loss of different aetiologies participated in the 6 month blinded phase of the study. Objective assessments showed a significant positive effect of treatment on hair growth. Intake of the active preparation for a further 6 months in an open phase indicated a subjective further improvement in hair growth. Exposure of the patients previously treated with placebo to the active preparation for 12 months gave similar results. Tolerability was good and no side-effects were reported. Nourkrin may provide an alternative to pharmacotherapy for the treatment of hair-loss problems in individuals with androgenetic alopecia. PMID- 17133781 TI - Interleukin-4, interleukin-8 and E-selectin levels in intranasal polyposis patients with and without allergy: a comparative study. AB - Intranasal polyposis is a chronic inflammatory upper airway disease with unknown aetiology. Interleukin (IL)-4 and IL-8 play very significant roles in allergic events and infectious inflammation, respectively. In contrast, E-selectin is important in the initiation and organization of inflammation. In this study, levels of IL-4, IL-8 and E-selectin were measured in nasal mucosa specimens of intranasal polyposis patients with and without allergic rhinitis, and healthy controls. Healthy controls had significantly higher IL-4 levels than allergic patients and non-allergic patients; IL-4 levels were similar in allergic and non allergic patients. Allergic and non-allergic patients had significantly higher IL 8 levels than healthy controls; IL-8 levels were comparable in allergic and non allergic patients. E-selectin levels were similar in all groups. The infection based theory, represented by IL-8, seems to be more likely than the allergy-based theory, represented by IL-4, for the pathogenesis of nasal polyposis. PMID- 17133782 TI - Acarbose, an alpha-glucosidase inhibitor, decreases aortic gene expression and serum levels of monocyte chemoattractant protein-1 in fructose-fed rats. AB - Insulin resistance is one of the determinants of post-prandial hyperglycaemia. Recently, acarbose, an alpha-glucosidase inhibitor that delays the absorption of carbohydrates from the small intestine, has been found to reduce the incidence of cardiovascular disease in patients with impaired glucose tolerance or diabetes. However, the molecular mechanism by which acarbose inhibits cardiovascular events remains unknown. In this study, we examined whether oral administration of acarbose could suppress expression of monocyte chemoattractant protein-1 (MCP-1) in fructose-fed rats, a widely used animal model of insulin resistance. Serum MCP 1 levels were elevated in fructose-fed rats after 4 weeks. Acarbose treatment for 4 weeks reduced the fructose-induced elevation of serum MCP-1 levels. Acarbose treatment for 8 weeks decreased MCP-1 mRNA levels in the aortae of fructose-fed rats. These results suggest that the cardioprotective effects of acarbose could be due, at least in part, to the suppression of MCP-1 expression. PMID- 17133783 TI - The effect of gynaecological laparoscopic surgery on cerebral oxygenation. AB - During gynaecological laparoscopic surgery, alterations in cerebral blood flow and intracranial pressure are frequently reported. These changes affect cerebral perfusion pressure and thus may affect cerebral oxygenation. In this prospective study, the effect of gynaecological laparoscopic surgery on cerebral oxygenation was examined by following the changes in regional cerebral oxygen saturation (rSo2). Twenty-four female patients were enrolled. The mean rSo2 was 65.5 +/- 5.4% at baseline before surgery, 60.8 +/- 5.6% when the patient was placed in the Trendelenburg position, 57.1 +/- 9.3% after creation of pneumoperitoneum, and 64.0 +/- 7.3% after the completion of surgery. During the period of pneumoperitoneum, rSo2 fell below 50% in two hypercapnic patients. In comparison with baseline, rSo2 declined significantly in the Trendelenburg position. The creation of pneumoperitoneum itself did not decrease the average rSo2 value further unless the patients were hypercapnic. PMID- 17133784 TI - Radiofrequency ablation of intra-articular osteoid osteoma of the hip. AB - Diagnosis and treatment of intra-articular osteoid osteoma is challenging. We present 16 patients with intra-articular osteoid osteomas of the hip treated with percutaneous radiofrequency ablation. Eight osteoid osteomas were located in the femoral head, six in the femoral neck, and two in the acetabulum. Three of the 16 patients had had an incorrect previous diagnosis. Percutaneous radiofrequency ablation was a clinical and technical success in all 16 patients. Within the first 24 h after the procedure, pain improved in all patients. Five patients had pain relief within the first 3 days after the procedure, nine patients within the first week and two patients within 2 weeks. Residual or recurrent symptoms were not reported by the last follow-up. At the 12-month follow-up, computed tomography and magnetic resonance imaging showed complete ossification and bone regeneration at the site of the lesion in three patients, partial ossification in six patients and no changes in seven patients. Computed tomography-guided percutaneous radiofrequency ablation is a simple, minimally invasive, safe and effective method for the treatment of most intra-articular osteoid osteomas. PMID- 17133785 TI - Effects of rosiglitazone and pioglitazone combined with metformin on the prothrombotic state of patients with type 2 diabetes mellitus and metabolic syndrome. AB - In this multicentre, randomized, double-blind, controlled, parallel-group trial, 103 patients with type 2 diabetes mellitus and metabolic syndrome were randomized to receive one of two thiazolidinediones--pioglitazone or rosiglitazone--in combination with 1500 mg/day of metformin, increasing up to 3000 mg/day, for 12 months. Anthropometric, metabolic, coagulation and fibrinolysis parameters were assessed at baseline and after 3, 6, 9 and 12 months. Significant decreases in glycosylated haemoglobin, fasting plasma glucose and post-prandial plasma glucose levels were seen after 9 and 12 months in both groups, and significant decreases in fasting plasma insulin and post-prandial plasma insulin levels were seen after 12 months in both groups. In both groups, improvement in the homeostasis model assessment index compared with baseline was obtained only after 12 months. Plasminogen activator inhibitor-1 levels were significantly lower in both groups after 12 months compared with baseline values. In patients with type 2 diabetes mellitus and metabolic syndrome, the combination of metformin plus thiazolidinediones improved glycaemic control and produced a slight but significant reduction in plasminogen activator inhibitor-1 levels. PMID- 17133786 TI - Placement of two types of spiral Z-stents at the bronchial carina for the treatment of terminal lung cancer--a new method. AB - Dumon Y-stents and Dynamic stents are used to treat carinal stenosis, but their placement severely impairs the expectoration of secretions, making frequent bronchoscopic aspiration necessary. We report here five patients with terminal lung cancer who had stenosis of the lower trachea and main bronchi treated using spiral Z-stents. A long tapered spiral Z-stent was placed in the lower trachea and one main bronchus, and a short straight spiral Z-stent in the contralateral main bronchus. No patients required bronchoscopic aspiration of secretions after stenting. Before stenting, all of the patients were severely dyspnoeic, requiring oxygen and having to sit in the orthopnoeic position. After stenting, the patients' dyspnoea improved, with one patient becoming ambulant without the need for oxygen support. These results suggest that the use of spiral Z-stents in stenosis of the tracheal carina in advanced lung cancer is effective in reducing the need for bronchoscopic aspiration and enhancing quality of life. PMID- 17133787 TI - Ethics and advertising. PMID- 17133788 TI - Dental implantology, who is trained to be an implantologist? PMID- 17133789 TI - Resolving the problem of balancing the profile of professionals in our country. PMID- 17133790 TI - Design of an x-ray room for a dental practice. AB - Radiography is one of the most valuable diagnostic tools used in comprehensive dental care. Although it is generally accepted that there is no safe level of radiation exposure, the possible risk involved with exposure to X-rays must be weighed against the benefits of clinical interpretation. One important step in limiting the risk of radiation exposure at the workplace is the correct design of an X-ray room. A dentist or dental therapist, who is the license holder of the radiation equipment, is ultimately responsible for the radiation safety at the workplace. The aim of this article is to provide dental practitioners with guidelines and recommendations on X-ray room designs in order to facilitate radiation control and safe working conditions for radiation workers as well as the public. PMID- 17133792 TI - An overview of bleaching techniques: history, chemistry, safety and legal aspects (part 1). PMID- 17133791 TI - The cleft seal for bottle-feeding--A report on case studies. AB - Clefting of the lip and palate is the most frequent craniofacial malformation that occurs in newborn babies. The greatest immediate challenge facing a neonate with a cleft is to take nourishment. In developing countries and particularly in rural areas, palatal obturators and other aids that can assist in feeding are not easily available or accessible. The aim of this study was to develop, evaluate and report on the cleft seal, a simple and inexpensive device that is used with a feeding bottle to promote nourishment of a neonate with a cleft palate. The cleft seal is made of either silicone or thermoform plastic and consists of a spoon like projection attached to a circular washer. The latter fits snugly between the teat base and collar of the feeding bottle, while the spoon-like projection forms a canopy over the teat and separates the nasal cavity from the mouth during feeding. Closure of the cleft emulates the natural feeding process and favours normal orofacial development. Results from this study indicate that there is an increase in nutritional intake over a shorter feeding time when using the cleft seal. Parents also report an absence of nasal regurgitation. The greatest advantage of the cleft seal lies in its negligible cost and ease of use, which can enable distribution and application in remote neonatal centres, where nursing and parental care givers are often at a loss as to what to do when faced with a newborn with a cleft palate. PMID- 17133793 TI - Necrotizing gingivitis of Kaposi sarcoma affected gingivae. AB - Necrotizing gingivitis and oral Kaposi sarcoma are common concomitants of HIV infection and both are regarded as indicators of HIV infection. Their simultaneous appearance in an HIV seropositive subject therefore, should be relatively common; but other reports documenting such cases could not be found. This article documents an uncommon case of necrotizing gingivitis superimposed on Kaposi sarcoma-affected gingiva, occurring in a patient with chronic periodontitis. The nature of necrotizing gingivitis and Kaposi sarcoma and the possible differential diagnosis of the periodontal attachment loss are discussed. PMID- 17133794 TI - General practitioner's radiology case 44. Compound odontoma. PMID- 17133795 TI - The atraumatic extraction. PMID- 17133796 TI - General practitioner's pathology case 7. Squamous cell carcinoma. PMID- 17133797 TI - Microbiological contamination of pig carcasses at different stages of slaughter in two European Union-approved abattoirs. AB - At sequential steps of slaughter (scalding, dehairing, singeing, polishing, trimming, washing, and chilling), 200 pig carcasses from two abattoirs were examined for total viable bacteria count (TVC) and the presence of Enterobacteriaceae and coagulase-positive Staphylococcus (CPS) by the wet-dry double-swab technique at the neck, belly, back, and ham. Before scalding, mean TVCs ranged from 5.0 to 6.0 log CFU cm(-2), and Enterobacteriaceae and CPS were detected on all carcasses. At abattoir A, mean TVCs and the percentage of Enterobacteriaceae-positive carcasses were reduced (P < 0.05) after scalding (1.9 log CFU cm(-2) and 12%, respectively), singeing (1.9 log CFU cm(-2) and 66%, respectively), and blast chilling (2.3 log CFU cm(-2) and 17%, respectively) and increased (P < 0.05) after dehairing (3.4 log CFU cm(-2) and 100%, respectively) and polishing (2.9 log CFU cm(-2)). The proportion of CPS-positive samples decreased to < or = 10% after scalding and remained at this level. At abattoir B, mean TVCs and the percentages of Enterobacteriaceae- and CPS-positive carcasses were reduced (P < 0.05) after scalding (2.4 log CFU cm(-2) and 29 and 20%, respectively), polishing (3.7 log CFU cm(-2)), and chilling (2.6 log CFU cm(-2) and 55 and 77%, respectively) and increased (P < 0.05) after the combined dehairing-singeing (4.7 log CFU cm(-2) and 97 and 100%, respectively). Among sites, the neck tended to yield higher levels of contamination from trimming to chilling at both abattoirs (P < 0.05). Consequently, scalding, singeing, and chilling may be integrated in a hazard analysis critical control point (HACCP) system for pig slaughter. As indicated by the higher levels of contamination on carcasses after dehairing-singeing and the following stages at abattoir B, each abattoir should develop its own baseline data and should customize HACCP systems to match process- and site-specific circumstances. PMID- 17133798 TI - Prevalence of Salmonella in diverse environmental farm samples. AB - The development of suitable intervention strategies to control Salmonella populations at the farm level requires reliable data on the occurrence and prevalence of the pathogen. Previous studies on Salmonella prevalence have focused on acquiring data from specific farm types and/or selected regions. The purpose of this study was to evaluate the distribution of this pathogen across a variety of farm types and regions in order to generate comparative data from a diverse group of environmental samples. Farm samples (n = 2,496) were collected quarterly from 18 different farms across five states (Tennessee, North Carolina, Alabama, California, and Washington) over a 24-month period. The participating farms included beef and dairy cattle operations, swine production and farrowing facilities, and poultry farms (both broiler chicken and turkey). The samples were analyzed for the presence of Salmonella by means of the U.S. Food and Drug Administration's Bacteriological Analytical Manual methods optimized for farm samples. Salmonella isolates were characterized by automated riboprinting. Salmonella serovars were recovered from 4.7% of all samples. The majority of positive findings were isolated from swine farms (57.3%). The occurrence of Salmonella was lower on dairy farms (17.9%), poultry farms (16.2%), and beef cattle farms (8.5%). The most commonly isolated serovar was Salmonella Anatum (48.4%), which was isolated notably more frequently than the next most common Salmonella serovars, Arizonae (12.1%) and Javiana (8.8%). The results of this study suggest that significant reservoirs of Salmonella populations still exist on swine production facilities and to a lesser extent in other animal production facilities. Data showed that the surrounding farm environment could be an important source of contamination. PMID- 17133799 TI - Microflora of minimally processed frozen vegetables sold in Gaborone, Botswana. AB - Two hundred samples of minimally processed, frozen, and prepacked potato chips, peas, corn, and a variety of combined vegetables from supermarkets in Gaborone, Botswana, were examined microbiologically. Determination of aerobic mesophilic plate count, aerobic psychrotrophic plate count, lactic acid bacteria, yeasts and molds, coliforms, Listeria spp., and Staphylococcus aureus were done. Chips had the lowest mean log values for all of the microorganisms enumerated except yeasts and molds. The mean log values for single vegetables ranged from 3.6 to 9.1, 3.4 to 8.9, 2.9 to 5.6, and 2.1 to 6.5 log CFU/ g aerobic mesophilic plate count, aerobic psychrotrophic plate count, lactic acid bacteria, and yeasts and molds, respectively. The microbial profiles of peas and corn were almost similar (P < 0.001). The mean values for combined vegetables were clustered within 4.6 and 5.4 and 4.2 and 5.2 log CFU/g aerobic mesophilic plate count and aerobic psychrotrophic plate count, respectively. All of the vegetables had a coliform population distribution ranging from 0 to < 10(4) most probable number per g. The predominant gram-negative bacteria isolated included members of Enterobacteriaceae and Pseudomonaceae (86.2%). Escherichia coli was not detected in all of the samples. The organisms isolated included those responsible for spoilage in frozen vegetables, namely Pseudomonas, Klebsiella, Corynebacterium, lactic acid bacteria, and Flavobacterium. The predominant lactic acid bacteria were Lactobacillus spp. (55.9%). Other spoilage organisms were yeasts, and Cryptococcus spp. (55.4%) was predominant. Pathogens, namely Listeria monocytogenes, were also isolated at a rate of 2 to 10%, of which 4% was from corn, 2% each from peas and country crop, and 10% from stir-fry. Bacillus cereus was also isolated and accounted for 7.7% of the microorganisms from corn. S. aureus was isolated from all of the vegetables. Enterotoxigenic strains were from corn, peas, mixed vegetables, and stir-fry, and all of them produced enterotoxin A. In addition, the isolates from stir-fry vegetables also produced enterotoxins B and C. The study reveals the presence of pathogens and emerging opportunistic pathogens in the ready-to-use or ready-to-eat vegetables. If E. coli is the only indicator for safety and acceptability, consumers may be exposed to foodborne diseases. Inclusion of other groups as indicator organisms is suggested. Retailers are urged to invest in standby generators to maintain the cold chain. PMID- 17133800 TI - Prevalence of potentially pathogenic Bacillus cereus in food commodities in The Netherlands. AB - Randomly selected food commodities, categorized in product groups, were investigated for the presence and number of Bacillus cereus bacteria. If positive, and when possible, five separate colonies were isolated and investigated for the presence of four virulence factors: presence of genes encoding three enterotoxins (hemolysin BL [HBL], nonhemolytic enterotoxin [NHE], and cytotoxin K) and the ability to produce cereulide. In addition, the presence of psychrotrophic and mesophilic signatures was determined. The genes for NHE are found in more than 97% of the isolates, those for HBL in approximately 66% of the isolates, and the gene for cytotoxin K in nearly 50% of the isolates. Significant associations between product groups and (combinations of) virulence factors were the relatively low percentage of isolates from the "flavorings" group containing genes encoding NHE and the higher-than-average occurrence of both the genes encoding HBL and NHE in the "pastry" group. Cereulide was produced by 8.2% of the isolates but only in combination with the presence of genes for one or more other virulence factors. Most isolates (89.9%) were mesophilic; minorities of the isolates were psychrotrophic (4.4%) or of intermediate signature (5.7%). In the product group "milk and milk products," the incidence of strains with psychrotrophic or intermediate signatures is significantly higher than in the other product groups. In the product groups "flavorings," "milk and milk products," "vegetable(s) and vegetable products," "pastry," and "ready-to-eat foods," a relatively high number of samples contain high numbers of B. cereus bacteria. Within the product group "ready-to-eat foods," the products containing rice and pasta show a relatively high incidence of high numbers of B. cereus bacteria. PMID- 17133801 TI - Salmonella and Shigella in freshly squeezed orange juice, fresh oranges, and wiping cloths collected from public markets and street booths in Guadalajara, Mexico: incidence and comparison of analytical routes. AB - A survey of the presence of Salmonella and Shigella in freshly squeezed orange juice and related samples was conducted in Guadalajara, Mexico. One hundred samples of freshly squeezed orange juice were collected from 49 street booths and 51 small food service establishments. In addition, 75 fresh orange samples, each consisting of five orange units, and 75 wiping cloths were collected from the same establishments from which juice had been collected. Salmonella was isolated from 14, 20, and 23% of samples of orange juice, orange surfaces, and wiping cloths collected from street vendors, while Shigella was isolated from 6, 17, and 5% of these samples. In general, the frequency of isolation of these pathogens in samples from juice serving establishments at public markets was significantly lower than that found among street vendors (P < 0.05). Salmonella enterica serotypes Agona, Typhimurium, and Anatum were found in orange juice, fresh oranges, and wiping cloth samples, while serotype Mexico was found on fresh oranges and in wiping cloths and serotypes Muenchen and Panama were found only in wiping cloth samples. Regarding Shigella species, Shigella sonnei was found in all three types of sample tested; Shigella dysenteriae was found in juice and orange samples, Shigella boydii in orange and wiping cloth samples, and Shigella flexneri on oranges only. Thirty-one percent and 39% of the juice samples showed aerobic plate counts of > or = 5.0 log CFU/ml and Escherichia coli counts of > 3.0 log CFU/ml, respectively. These high counts may indicate poor sanitation and potential exposure to fecal contamination either in the raw materials or during the orange-crushing and juice-serving process. These data may be useful for a further risk assessment of Salmonella or Shigella in unpasteurized, freshly squeezed juice. PMID- 17133802 TI - U.S. Food safety and Inspection Service testing for Salmonella in selected raw meat and poultry products in the United States, 1998 through 2003: an establishment-level analysis. AB - The U.S. Food Safety and Inspection Service (FSIS) pathogen reduction-hazard analysis critical control point systems final rule, published in 1996, established Salmonella performance standards for broiler chicken, cow and bull, market hog, and steer and heifer carcasses and for ground beef, chicken, and turkey meat. In 1998, the FSIS began testing to verify that establishments are meeting performance standards. Samples are collected in sets in which the number of samples is defined but varies according to product class. A sample set fails when the number of positive Salmonella samples exceeds the maximum number of positive samples allowed under the performance standard. Salmonella sample sets collected at 1,584 establishments from 1998 through 2003 were examined to identify factors associated with failure of one or more sets. Overall, 1,282 (80.9%) of establishments never had failed sets. In establishments that did experience set failure(s), generally the failed sets were collected early in the establishment testing history, with the exception of broiler establishments where failure(s) occurred both early and late in the course of testing. Small establishments were more likely to have experienced a set failure than were large or very small establishments, and broiler establishments were more likely to have failed than were ground beef, market hog, or steer-heifer establishments. Agency response to failed Salmonella sample sets in the form of in-depth verification reviews and related establishment-initiated corrective actions have likely contributed to declines in the number of establishments that failed sets. A focus on food safety measures in small establishments and broiler processing establishments should further reduce the number of sample sets that fail to meet the Salmonella performance standard. PMID- 17133803 TI - U.S. Food Safety and Inspection Service testing for Salmonella in selected raw meat and poultry products in the United States, 1998 through 2003: analysis of set results. AB - The U.S. Food Safety and Inspection Service (FSIS) tests sets of samples of selected raw meat and poultry products for Salmonella to ensure that federally inspected establishments meet performance standards defined in the pathogen reduction-hazard analysis and critical control point system (PR-HACCP) final rule. In the present report, sample set results are described and associations between set failure and set and establishment characteristics are identified for 4,607 sample sets collected from 1998 through 2003. Sample sets were obtained from seven product classes: broiler chicken carcasses (n = 1,010), cow and bull carcasses (n = 240), market hog carcasses (n = 560), steer and heifer carcasses (n = 123), ground beef (n = 2,527), ground chicken (n = 31), and ground turkey (n = 116). Of these 4,607 sample sets, 92% (4,255) were collected as part of random testing efforts (A sets), and 93% (4,166) passed. However, the percentage of positive samples relative to the maximum number of positive results allowable in a set increased over time for broilers but decreased or stayed the same for the other product classes. Three factors associated with set failure were identified: establishment size, product class, and year. Set failures were more likely early in the testing program (relative to 2003). Small and very small establishments were more likely to fail than large ones. Set failure was less likely in ground beef than in other product classes. Despite an overall decline in set failures through 2003, these results highlight the need for continued vigilance to reduce Salmonella contamination in broiler chicken and continued implementation of programs designed to assist small and very small establishments with PR-HACCP compliance issues. PMID- 17133804 TI - Distribution of virulent and pandemic strains of Vibrio parahaemolyticus in three molluscan shellfish species (Meretrix meretrix, Perna viridis, and Anadara granosa) and their association with foodborne disease in southern Thailand. AB - Distribution of pandemic strains of Vibrio parahaemolyticus in seafood, particularly in molluscan shellfish, and their serological and molecular relationships to clinical strains were examined from Hat Yai City in southern Thailand. During 2000 to 2002, virulent strains (tdh+ or trh+) were isolated from 13 of 230 molluscan shellfish samples using alkaline peptone water enrichment followed by immunomagnetic separation. The isolates included 12 pandemic strains (tdh+, trh-, group-specific PCR positive) from five Oriental hard clam samples, five green mussel samples, and one bloody clam sample. Among the pandemic strains, eight belonged to serogroup O3:K6, three belonged to O1:K25, and one was O1:K untypeable. One hundred eighty-seven strains of V. parahaemolyticus were isolated from clinical specimens obtained from a hospital in this city during 2000 to 2001. The pandemic strains comprised 64 and 68% of the isolates in 2000 and 2001, respectively. Among the serotypes of the pandemic strains, O3:K6 was dominant at 73% in 2000 and 76% in 2001 followed by O1:K25 at 20% in 2000 and 13% in 2001. Pulsed-field gel electrophoresis profiles of the pandemic strains from molluscan shellfish were indistinguishable or very similar to those of patient isolates. Similarity of the serotype distribution and DNA fingerprints occurring between the molluscan shellfish strains and clinical strains suggests that molluscan shellfish may be an important source of pandemic V. parahaemolyticus infection in southern Thailand. For public health, proper cooking of molluscan shellfish in this area is strongly recommended. PMID- 17133805 TI - Lethality of chlorine, chlorine dioxide, and a commercial produce sanitizer to Bacillus cereus and Pseudomonas in a liquid detergent, on stainless steel, and in biofilm. AB - Many factors that are not fully understood may influence the effectiveness of sanitizer treatments for eliminating pathogens and spoilage microorganisms in food or detergent residues or in biofilms on food contact surfaces. This study was done to determine the sensitivities of Pseudomonas cells and Bacillus cereus cells and spores suspended in a liquid dishwashing detergent and inoculated onto the surface of stainless steel to treatment with chlorine, chlorine dioxide, and a commercial produce sanitizer (Fit). Cells and spores were incubated in a liquid dishwashing detergent for 16 to 18 h before treatment with sanitizers. At 50 microg/ml, chlorine dioxide killed a significantly higher number of Pseudomonas cells (3.82 log CFU/ml) than did chlorine (a reduction of 1.34 log CFU/ml). Stainless steel coupons were spot inoculated with Pseudomonas cells and B. cereus cells and spores, with water and 5% horse serum as carriers. Chlorine was more effective than chlorine dioxide in killing cells and spores of B. cereus suspended in horse serum. B. cereus biofilm on stainless steel coupons that were treated with chlorine dioxide or chlorine at 200 microg/ml had total population reductions (vegetative cells plus spores) of > or = 4.42 log CFU per coupon; the number of spores was reduced by > or = 3.80 log CFU per coupon. Fit (0.5%) was ineffective for killing spot-inoculated B. cereus and B. cereus in biofilm, but treatment with mixtures of Fit and chlorine dioxide caused greater reductions than did treatment with chlorine dioxide alone. In contrast, when chlorine was combined with Fit, the lethality of chlorine was completely lost. This study provides information on the survival and sanitizer sensitivity of Pseudomonas and B. cereus in a liquid dishwashing detergent, on the surface of stainless steel, and in a biofilm. This information will be useful for developing more effective strategies for cleaning and sanitizing contact surfaces in food preparation and processing environments. PMID- 17133806 TI - A comparative study of two food model systems to test the survival of Campylobacter jejuni at -18 degrees C. AB - The survival of Campylobacter jejuni NCTC 11168 was tested at freezing conditions (-18 degrees C) over a period of 32 days in two food models that simulated either (i) the chicken skin surface (skin model) or (ii) the chicken juice in and around a broiler carcass (liquid model). In the skin model, cells were suspended in chicken juice or brain heart infusion broth (BHIB) and added to 4-cm2 skin pieces, which were subsequently stored at -18 degrees C. In the liquid model, cells were suspended in chicken juice or BHIB and stored at -18 degrees C. The decrease in the number of viable C. jejuni NCTC 11168 cells was slower when suspended in chicken juice than in BHIB. After freezing for 32 days, the reductions in the cell counts were 1.5 log CFU/ml in chicken juice and 3.5 log CFU/ml in BHIB. After the same time of freezing but when inoculated onto chicken skin, C. jejuni NCTC 11168 was reduced by 2.2 log units when inoculated in chicken juice and 3.2 log units when inoculated into BHIB. For both models, the major decrease occurred within the first 24 h of freezing. The results obtained in the liquid model with chicken juice were comparable to the reductions of Campylobacter observed for commercially processed chickens. The survival at -18 degrees C in the liquid model was also tested for three poultry isolates and three human clinical isolates of the serotypes 1.44, 2, and 4 complex. As observed for C. jejuni NCTC 11168, all the strains survived significantly better in chicken juice than in BHIB and were not notably influenced by serotype or origin. The findings indicate that the composition of the medium around the bacteria, rather than the chicken skin surface, is the major determining factor for the survival of C. jejuni at freezing conditions. The liquid model with chicken juice was therefore the best model system to study the freezing tolerance in Campylobacter strains. PMID- 17133807 TI - Growth and stress resistance variation in culture broth among Listeria monocytogenes strains of various serotypes and origins. AB - Twenty-five Listeria monocytogenes strains of various serotypes and sources, including clinical and food isolates associated with the same outbreaks, were characterized and compared based on growth rates and heat and acid death rates. Growth was monitored in tryptic soy broth supplemented with 0.6% yeast extract (TSBYE) at 4 and 30 degrees C for 32 days and 20 h, respectively. Heat and acid stress responses in TSBYE heated to 55 degrees C or acidified to pH 3.0 with lactic acid were evaluated for 240 or 120 min, respectively. Extensive variation in growth and stress resistance was observed among the tested strains. Growth rate differences were less evident at 30 than at 4 degrees C, where growth rates (log CFU per milliliter per day) ranged from 0.28 to 0.43. Thermal and acid death rates (log CFU per milliliter per minute) ranged from -0.023 to -0.052 and from 0.012 to -0.134, respectively. Serotype appeared to play a significant role (P < 0.05) only with respect to the heat resistance of the organism. Serotype 4b isolates as a group had lower heat resistance than did isolates representing all other serotypes combined. Although no clear origin-related (food versus clinical) trends were observed under the tested conditions, outbreak-related isolates of serotype 4b had lower acid death rates (higher acid resistance) (P < 0.05) than did the rest of the strains belonging to this serotype. Strain Scott A exhibited slow growth at 4 degrees C and low acid resistance, behavior that was distinct among both clinical and serotype 4b isolates. The results of this study highlight the risks associated with extrapolation to other strains of findings obtained with only one strain of L. monocytogenes. This information should be useful when test strains are to be selected for the evaluation of antimicrobial alternatives in ready-to-eat meat and other food products and when risk assessments are to be conducted. PMID- 17133808 TI - Quantifying nonthermal inactivation of Listeria monocytogenes in European fermented sausages using bacteriocinogenic lactic acid bacteria or their bacteriocins: a case study for risk assessment. AB - Listeria monocytogenes NCTC10527 was examined with respect to its nonthermal inactivation kinetics in fermented sausages from four European countries: Serbia Montenegro, Hungary, Croatia, and Bosnia-Herzegovina. The goal was to quantify the effect of fermentation and ripening conditions on L. monocytogenes with the simultaneous presence or absence of bacteriocin-producing lactic acid bacteria (i.e., Lactobacillus sakei). Different models were used to fit the experimental data and to calculate the kinetic parameters. The best model was chosen based on statistical comparisons. The Baranyi model was selected because it fitted the data better in most (73%) of the cases. The results from the challenge experiments and the subsequent statistical analysis indicated that relative to the control condition the addition of L. sakei strains reduced the time required for a 4-log reduction of L. monocytogenes (t(4D)). In contrast, the addition of the bacteriocins mesenterocin Y and sakacin P decreased the t(4D) values for only the Serbian product. A case study for risk assessment also was conducted. The data of initial population and t(4D) collected from all countries were described by a single distribution function. Storage temperature, packaging method, pH, and water activity of the final products were used to calculate the inactivation of L. monocytogenes that might occur during storage of the final product (U.S. Department of Agriculture Pathogen Modeling Program version 7.0). Simulation results indicated that the addition of L. sakei strains significantly decreased the simulated L. monocytogenes concentration of ready-to-eat fermented sausages at the time of consumption. PMID- 17133809 TI - Intragastric inoculation with a cocktail of Listeria monocytogenes strains does not potentiate the severity of infection in A/J mice compared to inoculation with the individual strains comprising the cocktail. AB - Although multistrain cocktails of Listeria monocytogenes are used in food inoculation experiments, no studies, to our knowledge, have been reported that use these cocktails in an intragastric mouse model. In this study, we used a five strain L. monocytogenes cocktail consisting of strains Scott A, MFS108, 101M, V7, and 310 and a four-strain L. monocytogenes cocktail containing strains Scott A, EGD, H7738, and F2365. Here, we report that intragastric inoculation of anesthetized mice with approximately 106 CFU of a cocktail of L. monocytogenes strains does not result (P > 0.05) in a more severe infection (on the basis of the CFU of Listeria spp. recovered from the spleen, liver, and blood) than inoculation of mice with similar numbers of the individual strains comprising the cocktail. Nor did we observe any consistent relationship between susceptibility of L. monocytogenes strains to inactivation in synthetic gastric fluid in vitro and virulence in mice. PMID- 17133810 TI - Effects of microbial inhibitors and modified atmosphere packaging on growth of Listeria monocytogenes and Salmonella enterica Typhimurium and on quality attributes of injected pork chops and sliced cured ham. AB - This study was designed to determine the inhibitory effects of modified atmosphere packaging (MAP), both alone and in combination with potassium lactate and sodium diacetate (PLSD), on selected pathogens common to pork products. Effects of the treatments on product quality also were assessed. The hypothesis was that high-CO2 MAP would increase the effectiveness of PLSD for inhibition of pork pathogens. Fresh chops from untreated pork loins and loins that were injected with PLSD were inoculated with Salmonella enterica Typhimurium, and slices of untreated hams and hams that were injected with PLSD were inoculated with Listeria monocytogenes. The samples were subjected to vacuum packaging (VP) or MAP with high concentrations (99.5 to 100%) of CO2. Pathogens were enumerated periodically during storage at 4 and 10 degrees C. Storage of pork chops at 4 degrees C slowed the growth of Salmonella Typhimurium, and there was little difference in pathogen numbers between VP and MAP samples. L. monocytogenes growth on ham slices at 4 degrees C was inhibited for up to 28 days by all of the treatments, but after 28 days, the VP-PLSD treatment had a greater inhibitory effect than did the other treatments. At 10 degrees C, the PLSD and MAP treatments each effectively inhibited the growth of the pathogens on pork chops and ham slices when compared with controls (VP). However, the results obtained with MAP plus PLSD at 10 degrees C were not different from those obtained with either MAP or PLSD alone. Therefore, the hypothesis was not supported; the high CO2 atmosphere of the MAP did not increase the effectiveness of PLSD for inhibition of pathogens. PMID- 17133811 TI - Survival, elongation, and elevated tolerance of Salmonella enterica serovar Enteritidis at reduced water activity. AB - Growing microorganisms on dry surfaces, which results in exposure to low water activity (a(w)), may change their normal morphology and physiological activity. In this study, the morphological changes and cell viability of Salmonella enterica serovar Enteritidis challenged to low a(w) were analyzed. The results indicated that exposure to reduced a(w) induced filamentation of the cells. The amount of filamentous cells at a(w) 0.94 was up to 90% of the total number of cells. Surviving filamentous cells maintained their membrane integrity after exposure to low a(w) for 21 days. Furthermore, cells prechallenged to low a(w), obtained with an ionic humectant, demonstrated higher resistance to sodium hypochlorite than control cells. These resistant cells are able to survive disinfection more efficiently and can therefore cause contamination of foods coming in contact with surfaces. This points to the need for increased attention to cleaning of surfaces in household environments and disinfection procedures in processing plants. PMID- 17133812 TI - Heat tolerance of Salmonella enterica serovars Agona, Enteritidis, and Typhimurium in peanut butter. AB - Recent large foodborne outbreaks caused by Salmonella enterica serovars have been associated with consumption of foods with high fat content and reduced water activity, even though their ingredients usually undergo pasteurization. The present study was focused on the heat tolerance of Salmonella enterica serovars Agona, Enteritidis, and Typhimurium in peanut butter. The Salmonella serovars in the peanut butter were resistant to heat, and even at a temperature as high as 90 degrees C only 3.2-log reduction in CFU was observed. The obtained thermal inactivation curves were upwardly concave, indicating rapid death at the beginning (10 min) followed by lower death rates and an asymptotic tail. The curves fitted the nonlinear Weibull model with beta parameters < 1, indicating that the remaining cells have a lower probability of dying. beta at 70 degrees C (0.40 +/- 0.04) was significantly lower than beta at 80 degrees C (0.73 +/- 0.19) and 90 degrees C (0.69 +/- 0.17). Very little decrease in the viable population (less than 2-log decrease) was noted in cultures that were exposed to a second thermal treatment. Peanut butter is a highly concentrated colloidal suspension of lipid and water in a peanut meal phase. We hypothesized that differences in the local environments of the bacteria, with respect to fat content or water activity, explained the observed distribution and high portion of surviving cells (0.1%, independent of the initial cell number). These results demonstrate that thermal treatments are inadequate to consistently destroy Salmonella in highly contaminated peanut butter and that the pasteurization process cannot be improved significantly by longer treatment or higher temperatures. PMID- 17133813 TI - Spread of a green fluorescent protein-tagged Pseudomonas putida in a water pipe following airborne contamination. AB - An aerosol of green fluorescent protein-tagged Pseudomonas putida, created during high-pressure water cleaning of a coupon colonized by a biofilm of the green fluorescent protein bacterium, contaminated the water supply of an experimental setup. The upward spread of P. putida in a vertical pipe of supply water was 4.3 cm/day. Results highlight that a water supply to a food plant can be contaminated by an aerosol of environmental flora, created in typical cleaning operations, and become a reoccurring source of contamination. A practical response that could be taken in a food plant is briefly discussed. PMID- 17133814 TI - Systematic environmental evaluations to identify food safety differences between outbreak and nonoutbreak restaurants. AB - Restaurants are important settings for foodborne disease transmission. The Environmental Health Specialists Network (EHS-Net) was established to identify underlying factors contributing to disease outbreaks and to translate those findings into improved prevention efforts. From June 2002 through June 2003, EHS Net conducted systematic environmental evaluations in 22 restaurants in which outbreaks had occurred and 347 restaurants in which outbreaks had not occurred. Norovirus was the most common foodborne disease agent identified, accounting for 42% of all confirmed foodborne outbreaks during the study period. Handling of food by an infected person or carrier (65%) and bare-hand contact with food (35%) were the most commonly identified contributing factors. Outbreak and nonoutbreak restaurants were similar with respect to many characteristics. The major difference was in the presence of a certified kitchen manager (CKM); 32% of outbreak restaurants had a CKM, but 71% of nonoutbreak restaurants had a CKM (odds ratio of 0.2; 95% confidence interval of 0.1 to 0.5). CKMs were associated with the absence of bare-hand contact with foods as a contributing factor, fewer norovirus outbreaks, and the absence of outbreaks associated with Clostridium perfringens. However, neither the presence of a CKM nor the presence of policies regarding employee health significantly affected the identification of an infected person or carrier as a contributing factor. These findings suggest a lack of effective monitoring of employee illness or a lack of commitment to enforcing policies regarding ill food workers. Food safety certification of kitchen managers appears to be an important outbreak prevention measure, and managing food worker illnesses should be emphasized during food safety training programs. PMID- 17133815 TI - Efficacy of Candida sake CPA-1 formulation for controlling Penicillium expansum decay on pome fruit from different Mediterranean regions. AB - The effectiveness of a formulated product of the yeast Candida sake CPA-1 for controlling postharvest diseases on pome fruits was demonstrated in laboratory, semicommercial, and commercial trials carried out in the major pome fruit producing region of the European Union. First, one wettable powder and seven liquid formulations were tested in laboratory trials that involved two varieties of apples and two varieties of pears. In all cases, an efficacy similar to that of fresh cells was demonstrated in the control of artificial Penicillium expansum infection. After these trials, the formulated product chosen for semicommercial and commercial trials was LF1, a liquid formulation that is particularly suitable for commercial applications. In semicommercial trials, LF1 showed a performance similar to fresh cells in most trials, and the population dynamics of both fresh and formulated cells were quite stable throughout the storage period. This indicates the high viability of C. sake CPA-1 in this formulation and the absence of adverse effects during the formulation of the product, which may significantly affect both its ability to grow on fruit and its antagonistic activity. We evaluated the control of natural infection after applying the formulated product in a commercial drencher in different packinghouses. A significant reduction in the incidence of diseases was observed with a recommended dose of around 10(7) CFU/ml when natural infections were greater than 1%. In general, large quantities of yeast were observed on the surface of unwounded fruits of different pome fruit cultivars. Moreover, populations of this biocontrol agent increased rapidly on fruit surfaces and remained quite stable for a long time under commercial storage conditions. Commercial practices used in packinghouses were therefore successfully applied for this formulated product. PMID- 17133816 TI - Optimization of a fluorescence polarization immunoassay for rapid quantification of deoxynivalenol in durum wheat-based products. AB - A fluorescence polarization immunoassay previously described for deoxynivalenol (DON) screening in wheat was optimized for the rapid quantification of DON in durum wheat kernels, semolina, and pasta. A background signal was observed in both spiked and naturally contaminated samples, strictly depending on the testing matrix. After subtracting the background DON level for durum wheat (0.27 microg of DON per g), semolina (0.08 microg of DON per g), and pasta (0.04 microg of DON per g), an accurate quantification of DON was possible at levels greater than 0.10 microg/g for all matrices. Average recoveries from spiked samples (0.25 to 1.75 microg/g) were 98, 102, and 101% for wheat, semolina, and pasta, respectively. Comparative analyses of 35 naturally contaminated durum wheat samples, 22 semolina samples, and 26 pasta samples performed by both the fluorescence polarization method and high-pressure liquid chromatography/immunoaffinity cleanup showed a good correlation (r > 0.995). The fluorescence polarization method showed better accuracy and precision with respect to the high-pressure liquid chromatography method and is suitable for the rapid and quantitative determination of DON in durum wheat-based products at levels foreseen by existing or coming international regulations. PMID- 17133817 TI - A detailed study of thermal decomposition, amalgamation/ atomic absorption spectrophotometry methodology for the quantitative analysis of mercury in fish and hair. AB - The analytical method for determining the concentration of mercury in fish by thermal decomposition, amalgamation/ atomic absorption spectrophotometry was thoroughly studied. Specific issues addressed were accurate modeling of instrumental response, the use of quartz and nickel boats, carryover effects, software limitations, and troubleshooting. The DMA-80 Direct Mercury Analyzer instrument was calibrated using a total of 22 points, and the resultant curves statistically analyzed. At minimum, second-order polynomials were required to adequately model the data. TORT-2 standard reference material was analyzed in both quartz and nickel boats and found to give equivalent performance in both types of vessels and well within the 95% confidence interval. DOLT-3 standard reference material also yielded values well within the 95% confidence interval, but the DORM-2 standard reference material did not. Carryover effects were found to be minimal with a new catalyst tube but increased with catalyst age. Blanks should be run after the analysis of high mercury content samples; however, when the catalyst has aged, two blanks are required to reduce apparent mercury signals to nominal blank values. Comparable results between thermal decomposition, amalgamation/atomic absorption spectrophotometry and cold-vapor atomic absorption spectrophotometry were demonstrated. The feasibility of using this instrument to analyze hair was also explored and found to be suitable. Software problems and limitations have been noted when attempting to implement a high-throughput methodology. Instrumental drift was found to be minimal when operated over long periods. Blank values can provide important diagnostic indicators. PMID- 17133818 TI - Occurrence of proteolytic activity and N-acyl-homoserine lactone signals in the spoilage of aerobically chill-stored proteinaceous raw foods. AB - Proteolytic pseudomonads dominate the spoilage flora of aerobically chill-stored proteinaceous raw foods. Proteolysis during spoilage of these food systems affects both food quality and the dynamics of the bacterial community because it increases the availability of nutrients to the community as a whole. Quorum sensing, or cell-cell signaling, is associated closely with ecological interactions among bacteria in mixed communities. The potential role of quorum sensing in proteolytic food spoilage was examined, based on the evaluation of N acyl-homoserine lactone (AHL) signal molecules. The occurrence of proteolytic activity and AHL signals was studied during spoilage of aerobically chill-stored ground beef, fish, chicken, and raw milk. Pseudomonads dominated the psychrotrophic flora, followed distantly by members of the Enterobacteriaceae. The growth of pseudomonads was correlated with the occurrence of proteolytic activity in all food systems. AHL concentration began increasing significantly only after the onset of proteolytic activity. Widely divergent AHL profiles were revealed by thin-layer chromatography analysis of the different food samples, and these profiles were likely determined by the undefined bacterial flora in these systems and by the characterized pseudomonads and Enterobacteriaceae. Although Hafnia alvei was a major component of the Enterobacteriaceae flora in all foods tested and a strong AHL producer, the signal molecules produced by H. alvei strain EB1 did not influence protease production by Pseudomonas fluorescens strain 395 in vitro. These results do not indicate any clear correlation between the overall detectable AHL signal molecules accumulated in the food samples and proteolytic activity. PMID- 17133819 TI - Survey on the hygienic status of plastic doors of a pig abattoir. AB - Three pig abattoirs in northeastern Germany were visited for examination of flexible plastic doors at the access to the chilling room. Swabs were taken from the front and back of the doors and were examined for aerobic plate count, Enterobacteriaceae, Salmonella, and Campylobacter. The aerobic plate count was within the range of 1.2 to 4.1 log CFU/cm2 on the front. On the back, between 0.9 and 3.5 log CFU/cm2 were obtained. In all three abattoirs, Enterobacteriaceae, Salmonella, and Campylobacter were detected. The highest contamination was found at the bottom of the door wings. Transfer of pathogens via contact of carcasses and doors must be considered possible. As a consequence, this contact surface should be integrated into the control program of cleaning and disinfection. PMID- 17133820 TI - Occurrence and density of vibrio parahaemolyticus in live edible crustaceans from markets in China. AB - Vibrio parahaemolyticus is a marine bacterium causing foodborne disease. Occurrence of the bacterium was investigated in six species of edible crustaceans available from markets in mainland China. The bacterium was detected in 22 of 45 whole-body, shell, and feces samples, including mitten crabs, which are supposed to be produced in freshwater ponds. The mean densities ranged from 2.8 log CFU/g in mitten crabs (Eriocheir sinensis) to 5.1 CFU/g in giant tiger prawns (Penaeus monodon). In hemolymph and muscle samples collected axenically, V. parahaemolyticus was detected in all of the prawns at a mean density of 2.6 log most probable number (MPN)/g, in two of five striped stone crabs (Charybdis feriatus) at a mean density of 1.1 log MPN/ml, and two of five mangrove mud crabs (Scylla serrata) at a mean density of 1.3 log MPN/ml. When six mitten crabs were collected from two freshwater ponds in China and were examined, V. parahaemolyticus was not detected. It seemed that cross-contamination occurred among live crustaceans at the markets. The results suggest that proper handling, storage, and cooking of these crustaceans will be necessary to lessen the risk of foodborne illness from V. parahaemolyticus. PMID- 17133821 TI - Morphological and physiological responses of Campylobacter jejuni to stress. AB - Under conditions of stress, cells of Campylobacter assume a coccoid shape that may be an evolutionary strategy evolved by the organism to enable survival between hosts. However, the physiology of Campylobacter as it devolves from spiral to coccoid-shaped morphology is poorly understood. In this study, conditions influencing the survival of Campylobacter jejuni ATCC 35921 in broth were determined. Cells in late log phase were resuspended in broth at 4 or 60 degrees C. The culturability of these cold- or heat-stressed cell suspensions was determined by spread plate counts and the activity of cells by the direct viable count technique and 5-cyano-2,3-ditolyltetrazolium chloride staining. C. jejuni changed form completely from culturable to viable but nonculturable cells (VBNC) within 25 days at 4 degrees C, and 15 min at 60 degrees C. Light microscopy of C. jejuni VBNC cells showed that the spiral-shaped cells became coccoid, and transmission electron microscopy of C. jejuni VBNC cells showed that the outer membrane was lost in aging cell suspensions. Furthermore, a limited proteomic study was carried out to compare C. jejuni proteins that exhibited increased or decreased synthesis on exposure to 60 degrees C. PMID- 17133822 TI - Assessment of environmental factors on Listeria monocytogenes Scott A inlA gene expression by relative quantitative Taqman real-time reverse transcriptase PCR. AB - Several virulence factors are involved in Listeria monocytogenes pathogenicity. L. monocytogenes internalins, particularly internalin A, are required for bacterial adhesion to and invasion of human intestinal epithelial cells. The expression of internalins is thus related to virulence. Identification of conditions involved in regulating the expression of L. monocytogenes virulence factors is essential for developing targeted strategies to control listeriosis incidence and improving therapeutic approaches. The primary aim of this study was to develop a quantitative real-time reverse transcriptase PCR platform to study the impact of environmental factors on L. monocytogenes Scott A virulence factor expression, particularly in potentially complex ecosystems. A Taqman PCR-based, rapid quantitative gene expression evaluation method was established with the L. monocytogenes ribosomal protein L4 encoding gene used as an internal standard. Our data suggest that inlA expression is influenced by food composition and temperature, indicating that certain food processing or storage conditions, such as the use of lactic and acetic acids at common storage temperatures, could affect the expression of L. monocytogenes virulence factor. PMID- 17133823 TI - Inactivation of the Crp/Fnr family of regulatory genes in Listeria monocytogenes strain F2365 does not alter its heat resistance at 60 degrees C. AB - A surprising facet of the Listeria monocytogenes genome is the presence of 15 genes that code for regulators in the Crp/ Fnr family and include the virulence regulator PrfA. The genes under the transcriptional control of these regulators are currently undetermined, with the exception of some genes controlled by the major virulence regulator PrfA. Using 12 strains of L. monocytogenes, each with an inserted gene cassette that interrupts and renders nonfunctional a different L. monocytogenes strain F2365 Crp/Fnr regulator, we heat challenged each strain at 60 degrees C with an immersed-coil heating apparatus, modeled the survivor data to calculate the underlying mean and mode of the heat resistance distribution for each strain, and compared the thermal tolerance of each mutant to the wild-type strain to determine if any of the Crp/Fnr mutants demonstrated altered heat tolerance. All 12 of the Crp/Fnr mutant strains tested had heat resistance characteristics similar to the wild-type strain (P > 0.05), indicating that mutations in these Crp/Fnr genes neither increased nor decreased the sensitivity of L. monocytogenes strain F2365 to mild heat. PMID- 17133824 TI - Surrogates for the study of norovirus stability and inactivation in the environment: aA comparison of murine norovirus and feline calicivirus. AB - Human noroviruses (NoVs) are the leading cause of food- and waterborne outbreaks of acute nonbacterial gastroenteritis worldwide. As a result of the lack of a mammalian cell culture model for these viruses, studies on persistence, inactivation, and transmission have been limited to cultivable viruses, including feline calicivirus (FCV). Recently, reports of the successful cell culture of murine norovirus 1 (MNV-1) have provided investigators with an alternative surrogate for human NoVs. In this study, we compared the inactivation profiles of MNV-1 to FCV in an effort to establish the relevance of MNV-1 as a surrogate virus. Specifically, we evaluated (i) stability upon exposure to pH extremes; (ii) stability upon exposure to organic solvents; (iii) thermal inactivation; and (iv) surface persistence under wet and dry conditions. MNV-1 was stable across the entire pH range tested (pH 2 to 10) with less than 1 log reduction in infectivity at pH 2, whereas FCV was inactivated rapidly at pH values < 3 and > 9. FCV was more stable than MNV-1 at 56 degrees C, but both viruses exhibited similar inactivation at 63 and 72 degrees C. Long-term persistence of both viruses suspended in a fecal matrix and inoculated onto stainless steel coupons were similar at 4 degrees C, but at room temperature in solution, MNV-1 was more stable than FCV. The genetic relatedness of MNV-1 to human NoVs combined with its ability to survive under gastric pH levels makes this virus a promising and relevant surrogate for studying environmental survival of human NoVs. PMID- 17133825 TI - Comparison of the reveal test, the U.S. Food and Drug Administration culture method, and selective media for recovery of Salmonella enteritidis from commercial egg layer flock environments. AB - Salmonella is the leading cause of foodborne illnesses in the United States, and Salmonella Enteritidis (SE) is the second most frequently isolated Salmonella serovar. Egg products are most often associated with outbreaks of SE infection. To prevent SE contamination of eggs, many producers are implementing flock inspections for SE at their facilities. A rapid and simple method for detecting SE in poultry environmental samples is critical for effective control of SE. In this study, the Reveal test for SE was compared with the conventional U.S. Food and Drug Administration (FDA) culture method for detecting SE in naturally contaminated environmental samples. The efficacy of two enrichment media, tetrathionate broth (TT) and Rappaport-Vassiliadis medium (RV), and three selective plating media, brilliant green agar with novobiocin (BGN), xylose lysine tergitol 4 agar (XLT4), and bismuth sulfite agar (BS), also were compared for SE isolation. One hundred twenty-eight environmental drag swab samples were collected from two previously identified SE-positive chicken flocks in two U.S. states and analyzed in parallel using the Reveal test and the FDA culture method. Twenty-five samples (19.5%) yielded SE when the Reveal test was used, and 23 samples (18.0%) were positive for SE by the FDA culture method. No significant difference in efficacy (P = 0.527) was found between the two methods. The Reveal test had a sensitivity, specificity, and accuracy of 83, 94, and 92%, respectively. Overall, a significantly greater number of positive samples was obtained after enrichment in RV compared with TT. XLT4 and BGN were more efficient than BS for isolating SE. However, no single method or medium successfully recovered SE from all SE-positive environmental samples. PMID- 17133826 TI - Evaluation of an alkaline phosphatase-labeled oligonucleotide probe for the detection and enumeration of the thermostable-related hemolysin (trh) gene of Vibrio parahaemolyticus. AB - Reliable methods are needed to detect total and pathogenic Vibrio parahaemolyticus. One marker of V. parahaemolyticus virulence is the thermostable related hemolysin. We developed an alkaline phosphatase-labeled DNA probe method for the specific detection and enumeration of trh-positive V. parahaemolyticus by colony hybridization. The probe was tested against a panel of 200 bacterial strains and determined to be specific for trh-positive V. parahaemolyticus. Additionally, the trh alkaline phosphatase probe colony hybridization was successfully used to detect and enumerate trh-positive V. parahaemolyticus in seafood and water samples collected from the United States and the United Kingdom. PMID- 17133827 TI - Measurement of T-2 and HT-2 toxins in eggs by high-performance liquid chromatography with fluorescence detection. AB - T-2 toxin is a mycotoxin produced by several species of common fungi capable of infesting human food and animal feeds. Lower-quality feeds given to chickens may be contaminated with T-2 toxin, which may affect their health. The literature suggests that T-2 toxin is transmitted from the hen to the eggs. This article describes the development of a liquid chromatographic assay for T-2 and the related mycotoxin HT-2 in eggs. T-2 and HT-2 toxins were isolated from spiked eggs with a tandem charcoal-alumina-Florisil column and immunoaffinity column cleanup. The isolated toxins were derivatized with the fluorophore 1-anthroyl nitrile, separated by high-performance liquid chromatography, and quantitated by fluorescence. The limit of detection of the method was 1 ng ml(-1) (parts per billion) of T-2 and HT-2 in whole (with shell removed) eggs. The limit of quantitation for both toxins was 5 ng ml(-1). Recoveries from spiked eggs over the range from 5 to 50 ng ml(-1) averaged 89.2% for T-2 and 100.3% for HT-2, with coefficients of variation of 3.5 and 8.2%, respectively. This method is sensitive enough to be used to check for the presence of T-2 or HT-2 toxins in eggs. PMID- 17133828 TI - Fish meal in animal feed and human exposure to persistent bioaccumulative and toxic substances. AB - Persistent and bioaccumulative toxic substances (PBTSs) that end up in fish are health hazards and the object of fish-consumption advisories. Some of these substances are present as extraneous contaminants, e.g., man-made lipophilic pollutants such as organohalogen pollutants, and others such as monomethyl mercury can be considered naturally occurring. Omnivores (e.g., poultry and swine) and especially ruminants that are fed contaminated fish meal can pass monomethyl mercury and organohalogen pollutants to eggs, meat, and dairy products. Differences in fish meal PBTS profiles and farm animal (e.g., poultry, swine, cattle, and farmed fish) physiology modulate PBTSs in animal products. Fish-consumption advisories issued to protect human health do not extend to fish by-products fed to farmed animals. Animals (especially farmed fish) that are fed fish meal can bioconcentrate monomethyl mercury in protein matrices, and organohalogen pollutants can be passed on in the fat components of derived foods. Policies to decrease exposure to monomethyl mercury and organohalogen pollutants must consider farming practices that use fish by-products. A risk assessment of toxic contaminants in fish meal may indicate that food safety objectives must consider the human health impact of foods derived from animals fed contaminated meal. PMID- 17133829 TI - Inactivation of protozoan parasites in food, water, and environmental systems. AB - Protozoan parasites can survive under ambient and refrigerated storage conditions when associated with a range of substrates. Consequently, various treatments have been used to inactivate protozoan parasites (Giardia, Cryptosporidium, and Cyclospora) in food, water, and environmental systems. Physical treatments that affect survival or removal of protozoan parasites include freezing, heating, filtration, sedimentation, UV light, irradiation, high pressure, and ultrasound. Ozone is a more effective chemical disinfectant than chlorine or chlorine dioxide for inactivation of protozoan parasites in water systems. However, sequential inactivation treatments can optimize existing treatments through synergistic effects. Careful selection of methods to evaluate inactivation treatments is needed because many studies that have employed vital dye stains and in vitro excystation have produced underestimations of the effectiveness of these treatments. PMID- 17133830 TI - [Cold intolerance syndrome--a review]. AB - Cold sensitivity is defined as a complex of symptoms occurring after injuries, which patophysiology remains obscure, but is probably of multifactorial nature, and which may be a cause of severe complaints and disability of the hand when exposed to the cold. The symptoms include numbness and stiffness of fingers, pain, discolouration (paleness, cyanosis), swelling and reduction of grip strength. This disorder affects more frequently upper than lower extremity, may develop after variety of injuries and elective operations, but the most commonly occurs in patients with replanted fingers or hands. Their incidence is reported of 50-100% and is related to many factors, including mechanism of trauma, type of damaged structures, but also to climate in which patient lives and works. The symptoms tend either to disappear or exacerbate with time passing. There are no known measures or therapies which can prevent development of the syndrome or which can effectively manage it. PMID- 17133831 TI - [Lower extremity nerves injuries in bone fractures and joint dislocations- etiopathogenesis, treatment]. AB - Lower extremity nerve injuries accompanying bone fractures and joint dislocations lead to serious disturbances at the lower limbs and caused significant prolongation of the treatment time. Clinical material consisted of 73 patients treated surgically with lower extremity nerve injuries due to bone fractures and joint dislocation. Based on this material we analysed the most frequent localisations of the nerves lesions and the mechanism of injury. The indications to surgical treatment were also described. PMID- 17133832 TI - [Results of the operative treatment in Madelung's deformity]. AB - Nine wrists in eight patients were treated surgically between 1988 and 2003 for symptomatic Madelung's deformity. The pain of involved wrist followed by forearm deformation fulfilled criteria for surgery. Closing wedge osteotomy of the distal radius were carried out eight times accompanied by shortening of the ulna (four patients), excision of the distal ulna (one patient), and no ulnar surgery (three patients). Pain relieved after surgery. The follow up period ranged from 1 to 9.5 years. No pour results were stated in subjective patient's estimation during final check up. Wrist appearance were stated to be satisfactory. Limitation of the range of motion concerning supination and pronation of the forearm were stated invariably. X-ray retrospective assessment of the inclination angle, lunate coverage and presence of arthritic changes were conducted. Time and method of surgical treatment for Madelung's deformity should be considered individually. PMID- 17133833 TI - [Intraarticular distal radius fractures--arthroscopic assessment of injuries]. AB - Intraarticular distal radius fractures affect predominantly young people by contrary to "loco typico" fractures concerning old patients with osteoporosis. They are usually caused by high energy trauma and their treatment needs anatomic reposition. Between August and December 2004 there were performed 10 wrist arthroscopies in distal intraarticular radius fractures: 3 women and 7 men, age from 20 to 63 years, average 41. In all cases arthroscopy revealed larger displacement than assessed by standard radiographs and multiple associated soft tissue injuries were detected: lesions of TFCC in 5 patients, cartilage lesions of carpal bones in 6 and I degree SL interosseous ligament lesion in 4 patients. Arthroscopy in distal radius fractures allows detailed articular surface displacement assessment and reposition under "eye control". Additionally associated soft tissue injuries, which are not visible in standard X-ray examination are detected and treated. PMID- 17133834 TI - [Analysis of articular cartilage lesions in 5114 knee arthroscopies]. AB - The cartilage lesions become the increasing social problem, which occurs often in young people. It influences the normal knee function and may predispose patients for osteoarthritis. The aim of this study was the epidemiologic and etiological analysis of the knee articular cartilage lesions diagnosed in arthroscopy. From 1997 to 2002, arthroscopy of the knee joint was performed in 5114 patients. Chondral lesions were found in 2931 cases--57.3. Non-isolated cartilage lesions accounted for 67.9%. Grade II according to ICRS classification evaluated in arthroscopy was the most frequent grade of the cartilage lesion (47.8%) and grade 4B the least frequent one (0.11%). The patellar articular surface (43.3%) and the medial femoral condyle (39.6%) were the most common locations of the lesions. PMID- 17133835 TI - [The estimation of the results of the treatment patients with failed back surgery syndrome]. AB - The authors present the reasons failed lumbar disc operations among 51 patients, 53% of them were womae and 47% were mae. Age of the patients was from 30 to 73 years (average 49 years). Most of them have had some operations. They complained of constant, strong lumbar pain with sciatic pain, restriction of lumbar spine mobility and paresis. The results were compared after 12 months from final operation with applying 3 methods: decopmpression, transpedicular fixation with decompression and decompession conected with transpedicular fixation and inerbody fusion. Intraoperatively considerable amount of instability (60%) was found. The best results of treatment was gained in decompression conected with transpedicular fixation and interbody fusion. The good result came up 41% in this group and in the other 20%. PMID- 17133836 TI - [Hip joint development after treatment the developmental dislocation with overhead traction]. AB - Despite the fact that ultrasound of children's hips is widely used for screening, late diagnosed cases of developmental dislocation of the hip are still a common problem in the orthopaedic practice. The aim of the study is to review final clinical and radiological outcomes of treatment of DDH with overhead traction and closed reduction after skeletal growth. Clinical records and radiograms of 107 hips (81 children) were retrospectively reviewed. All of them were treated according to the same program: overhead traction (about 2 weeks), followed by closed reduction, modified Lorenz cast (2 months) and finally cast in Lange position (3 months). Average age of children was 14.2 months at the beginning of treatment and 20.7 years at last visit. Good and very good results were found in 80% of cases according to final radiological assessment of Severin and in 91% according functional classification of Harris. Avascular necrosis of femoral head according to Bucholz-Ogden classification system was identified in one third of patients. Functional results are better than radiological, but deteriorated with time especially in hips with residual dysplasia and AVN due to development of early, secondary degenerative changes. PMID- 17133837 TI - [PDT photodynamic therapy in orthopedic inflammatory conditions?]. AB - The infections in orthopedic surgery and traumatology represents important medical problem. The results of treatment of motor-organs infections in high degree were improved by the introduction of antisepsis and asepsis in XIX century and the antibiotics' discovery in the beginning of XX century. However, widespread usage of antibiotics leaded to the rise of refractory tribes of bacteria on their activity, which caused higher percentage of fails in the therapy. Also the rapid civilization development, which flown on arising of a new invasive methods of operative trauma protection and stabilizations of fractures in motor-organs area increased the number of infectious complications during treatment. These facts gave a reason to look for a more effective therapeutic methods. It seems that photodynamic therapy gives us the new possibilities of infectious treatment, which avails oneself activity of therapeutic light laser with proper wave length on the human tissue with photosensitizer included. Indeed there are known until now splendid results of PDT in neoplasma treatment, however it seems, based on experimental investigations and publications in medical literature, that this method can be a chance of effectual and non-invasive treatment of the soft tissues and bones inflammations. PMID- 17133838 TI - [Does coexistant fractures impair the results of treatment of elbow dislocations in children?]. AB - We present 33 children with 34 elbow dislocations. In 22 cases concomittant fractures around the elbowe were noted: 13 displaced medial epicondyle fractures, 5 radial neck fractures, 2 radial head fractures, 2 coronoid process fractures, 1 olecranon fracture, 1 lateral condyle fracture, 1 medial condyle fracture. In 5 children more than one concomittant fracture were noted. The mean age during dislocation was 11 years (from 4 to 15), the mean follow up period was 3.5 years (from 2 to 5). On follow up a clinical examination of the elbow was performed including axis assesment and elbow ROM. X-ray were taken in all children in AP and lateral view. All children had a satisfying function of the extremity and no pain was noted. No difference in ROM was noted in children with isolated elbow dislocation or with concomittant medial epicondyle fractures however in 3 of those children a persistent flexion contracture was observed. Among children with other fractures poorer results occured in 3 patients, specially after displaced radial neck fractures. In one of them there was slight supination and pronation restriction, in second a complete ankylosis of proximal radioulnar joint occured in neutral forearm position. In one child after elbow dislocation and medial condyle fracture a late radial dislocation occured. We think that concomitant medial epicondyle fracture does not impare good results of elbow dislocation treatment if well recognised and treated. Other fractures, specially radial neck fractures, increase the risk of complications. PMID- 17133839 TI - [Arthroscopy scapulothoracic articulation (introduction of this method)]. AB - The authors have introduced arthroscopy of scapulothoracic. They have treated 37 years old patient who suffer from bursitis with pain and crepitus. They have used arthroscopy procedure. The first publication about scapulothoracic arthroscopy was in 1992. The method has been written very rarely by foreigner authors in medical journals. PMID- 17133840 TI - [Ganglion cyst of the distal forearm mimicking a malignant tumour--a case report]. AB - A case of tumour localized in the distal forearm is described. CT imaging of the lesion mimicked a malignancy because of it's heterogeneous structure. Primary intended for open diagnostic biopsy, intraoperatively a lesion was recognized as a polycystic ganglion growing from the wrist joint. The tumour was resected completely and histopathology confirmed its benign nature. Authors touch the diagnostic difficulties in atypical clinical presentations of the ganglions and when they mimic a malignancy at imaging. PMID- 17133841 TI - [Out-patients unit--recapitulation of five-year activity]. AB - Authors make recapitulation of out-patients unit activity. This unit is a part of Department Physiotherapy, Rheumatology and Rehabilitation University of Medical Sciences in Poznan. Numbers of treated patients in following years, most frequent reasons of treatment and preferences of treatment among patients are given. Finally, authors put particular accent on still growing up interest in this form of rehabilitation activity. PMID- 17133842 TI - [The results of surgical treatment of tibial plateau fractures]. AB - Between 1995-2003 84 patients were treated because of the tibial plateau fracture in the Department of Orthopaedics of Gdansk Medical University. 47 patients were evaluated at the follow-up. In the pure cleavage fractures after joint opening an open reduction and internal fixation via screws or buttress plate was performed. In the cleavage fractures combined with depression the treatment consisted of: open reduction, elevation of the depressed plateau "en mass", bone grafting of the metaphysis, fixation of the fracture with cancellous screws and buttress plating. Rasmussen modified scale was used to assess the outcome. Resnick and Niwoyama criteria were used in calculating the degree of the degenerative changes. The clinical results were very good in 5 patients, good in 26, fair in 9 and poor in 5 patients. In the radiological assessment a very good result were observed in 8 patients, good in 28, fair in 7 and poor in 4 patients. An anatomical reduction of the articular surface together with stable internal fixation of the fracture site decreases the risk of the degenerative changes and helps to achieve a good final result. PMID- 17133843 TI - Drug-eluting stents--safety concerns. PMID- 17133844 TI - Nitric oxide synthase inhibitors in post-myocardial infarction cardiogenic shock- an update. AB - Cardiogenic shock (CS) in acute myocardial infarction, after successful coronary angioplasty, still carries a case fatality rate of 50%. These patients succumb to a systemic metabolic storm, superimposed on extensive myocardial necrosis and stunning. Nitric oxide (NO) overproduction contributes to the pathophysiology of this morbid state. Current data regarding the physiologic effects of NO and nitric oxide synthase (NOS) inhibitors on the cardiovascular system are reviewed. Clinical trials assessing the safety and efficacy of NOS inhibitors in CS are summarized. PMID- 17133845 TI - Coronary anomalies by cardiac computed tomographic angiography. AB - BACKGROUND: Early identification of an anomalous coronary anatomy is quite relevant because of the increased incidence of sudden cardiac death or related symptoms of myocardial ischemia in the patients. Invasive coronary angiography (ICA) is not only invasive, but expensive, and cannot always adequately provide the required information about the abnormal coronary anatomy. Cardiac computed tomographic angiography (CTA) is a robust noninvasive imaging modality that has several clinical applications and is now being used increasingly in practices across the nation. It not only provides high-resolution anatomical information of the coronary artery tree but also helps define other aspects of the cardiovascular anatomy, be it normal or abnormal. HYPOTHESIS: This study sought to determine the clinical role played by CTA in the evaluation of different types of coronary arterial anomalies by reviewing CTA studies since 1997. METHODS: We reviewed 6,089 case studies of contrast CTA conducted at our institution. There were 53 coronary anomalies in 39 patients (0.64%). RESULTS: Computed tomographic angiography correctly identified the course of coronary arteries in all cases. CONCLUSION: The results of this study support the use of CTA as a safe and effective noninvasive imaging modality for defining coronary arterial anomalies in an appropriate clinical setting, providing detailed three-dimensional anatomic information that may be difficult to obtain with invasive angiography. PMID- 17133846 TI - The effect of coronary angiography on residual renal function in patients on peritoneal dialysis. AB - BACKGROUND: The risk of intravascular radiocontrast to residual renal function (RRF) in patients on peritoneal dialysis (PD) remains largely unknown. HYPOTHESIS: This study sought to estimate the effect of coronary angiography on RRF in patients on PD. METHODS: All patients at the VA Pittsburgh Healthcare System and University of Pittsburgh who underwent coronary angiography between 1993 and 2005 while on PD and who had RRF measured prior to angiography were identified retrospectively. For patients without a postprocedure RRF recorded, medical records were reviewed to determine whether anuria had developed. The longer-term rate of loss of RRF among cases was compared with a composite rate of decline in RRF among cases before angiography and matched controls. RESULTS: Twenty-nine patients with a mean preprocedure RRF of 4.4+/-3.2 ml/min/1.73m(2) were evaluated. Of these patients, 23 (79%) had postangiography RRF assessments (mean clearance 3.4+/-3.0 ml/min/1.73m(2)). One of the remaining six patients definitely became permanently anuric following angiography, one was lost to follow-up, and there was no postprocedure RRF assessment in four others. The rate of decline in RRF in the cases was similar to the composite rate (0.07 ml/min/1.73m(2)/month vs. 0.09 ml/min/1.73m(2)/month, p=0.53) CONCLUSION: The risk for permanent anuria in patients on PD undergoing coronary angiography appears to be quite small. Patients who do not develop anuria following coronary angiography have the same gradual rate of loss of RRF as other patients on PD. Providers should be vigilant in protecting RRF in patients on PD undergoing coronary angiography. PMID- 17133847 TI - Effect of concordance between sites of left ventricular pacing and dyssynchrony on acute electrocardiographic and echocardiographic parameters in patients with heart failure undergoing cardiac resynchronization therapy. AB - BACKGROUND: Wide QRS complexes are associated with left ventricular (LV) dyssynchrony, but an optimal site of LV pacing has not been established. HYPOTHESIS: We hypothesized that concordance between the sites of LV pacing and of latest LV mechanical activation during cardiac resynchronization therapy (CRT) is associated with more favorable acute echocardiographic changes. METHODS: An analysis of 28 consecutive patients (64+/-15 years, 46% male, 52% ischemic heart disease, LV ejection fraction 0.24+/-0.07, QRS 163+/-22 ms, New York Heart Association > or = 3 93%) implanted with biventricular (BIV) defibrillators was performed. Sites of latest LV activation were determined by tissue Doppler imaging (TDI) and speckle tracking (ST). The site of LV pacing was determined by fluoroscopy in two views. A concordance score (0-5) was created to describe the proximity of the pacing site to the site of the latest mechanical activation. RESULTS: Compared with the worst concordance score, a perfect score was associated with shorter QRS width with LV (187+/-40 vs. 246+/-8 ms, p= 0.048) and BIV (134+/-19 ms vs. 179+/-39 ms, p = 0.05) but not with right ventricular pacing. A perfect concordance score was also associated with a greater acute reduction in LV volumes in systole (42+/-36 ms 16+/-22 ms, p = 0.068) and diastole (47+/-37 vs. 8+/-31 ml, p = 0.043) 24 h after CRT device implantation. CONCLUSIONS: A high concordance is associated with shorter QRS width with LV and BIV pacing and greater acute reduction in LV volumes. The effect of concordance on the intermediate and long-term response to BIV pacing deserves further evaluation. PMID- 17133848 TI - Clinical characteristics and outcomes of Chinese patients with cholesterol crystal embolism after coronary intervention. AB - BACKGROUND: Cholesterol crystal embolism (CCE) is a complication of atherosclerosis. Vascular surgery, vascular angiography, and anticoagulation have been identified as inciting factors. HYPOTHESIS: This paper sought to analyze the clinical characteristics of patients with CCE after percutaneous coronary intervention. METHODS: Six patients with atherosclerosis presenting with simultaneous occurrence of acute renal failure and peripheral ischemic changes were diagnosed with CCE and their clinical data were analyzed. RESULTS: The average age of the patients was 72 years. Most had risk factors of atherosclerosis such as hypertension, diabetes, and smoking. The levels of serum creatinine increased progressively after coronary angiography. All patients had concomitant skin lesions, including blue toes. Cholesterol crystal emboli were found in arterioles by cutaneous biopsy in one patient. All patients received statins and two of these received dialysis therapy. Three patients died and three remained in chronic renal failure. CONCLUSION: Since CCE is a severe complication of coronary intervention, special attention should be paid to this disease. PMID- 17133849 TI - Acute improvement of aortic mechanics following hemodialysis in patients with chronic renal failure. AB - BACKGROUND: Evidence suggests that distensibility of the aorta is decreased in patients with end-stage renal failure, while the underlying mechanisms are unclear. HYPOTHESIS: The purpose of the study was to evaluate the distensibility of the aorta in patients at the end stage of chronic renal failure before and after hemodialysis (HD). METHODS: The diameter of the ascending aorta and distensibility were assessed in 48 patients on HD (31 men, 17 women, aged 45+/-14 years) and in 27 normal subjects (17 men, 10 women, aged 44+/-14 years). The diameter of the aorta was evaluated by M-mode in the parasternal long-axis view. RESULTS: Aortic distensibility was significantly lower in patients on HD before HD (1.9+/-0.7 cm(2) x dyn(-1) x 10(-6)) than in normal control subjects (3.8+/ 1.0 cm(2) x dyn(-1) X 10(-6), p< 0.0001). After dialysis, it increased to 2.6+/ 1.2 (p < 0.05 compared with baseline, p < 0.001 compared with controls). The change of aortic distensibility correlated with age (R(2) = 0.629 p < 0.001) and ultrafiltration volume (R(2) = 0.168, p < 0.01). CONCLUSIONS: Aortic distensibility in patients with end-stage renal disease is significantly lower than in normal subjects, and it is significantly improved after HD. PMID- 17133850 TI - Images in cardiology: ruptured congenital sinus of Valsalva aneurysm. PMID- 17133851 TI - Images in cardiology: syncope: a case of vein graft rupture. PMID- 17133852 TI - Images in cardiology: ventriculo-atrial Wenckebach during wide complex tachycardia. PMID- 17133853 TI - Theodor Wilhelm Engelmann. PMID- 17133854 TI - The evolution of female mating preferences: differentiation from species with promiscuous males can promote speciation. AB - Females of many species are frequently courted by promiscuous males of their own and other closely related species. Such mating interactions may impose strong selection on female mating preferences to favor trait values in conspecific males that allow females to discriminate them from their heterospecific rivals. We explore the consequences of such selection in models of the evolution of female mating preferences when females must interact with heterospecific males from which they are completely postreproductively isolated. Specifically, we allow the values of both the most preferred male trait and the tolerance of females for males that deviate from this most preferred trait to evolve. Also, we consider situations in which females base their mating decisions on multiple male traits and must interact with males of multiple species. Females will rapidly differentiate in preference when they sometimes mistake heterospecific males for suitable mates, and the differentiation of female preference will select for conspecific male traits to differentiate as well. In most circumstances, this differentiation continues indefinitely, but slows substantially once females are differentiated enough to make mistakes rare. Populations of females with broader preference functions (i.e., broader tolerance for males with trait values that deviate from females' most preferred values) will evolve further to differentiate if the shape of the function cannot evolve. Also, the magnitude of separation that evolves is larger and achieved faster when conspecific males have lower relative abundance. The direction of differentiation is also very sensitive to initial conditions if females base their mate choices on multiple male traits. We discuss how these selection pressures on female mate choice may lead to speciation by generating differentiation among populations of a progenitor species that experiences different assemblages of heterospecifics. Opportunities for differentiation increase as the number of traits involved in mate choice increase and as the number of species involved increases. We suggest that this mode of speciation may have been particularly prevalent in response to the cycles of climatic change throughout the Quaternary that forced the assembly and disassembly of entire communities on a continentwide basis. PMID- 17133855 TI - Epistasis and dominance: evidence for differential effects in life-history versus morphological traits. AB - Dominance and epistatic effects are predicted to be larger in life-history than in morphological traits. We test these predictions using published results from line cross analyses. We find that dominance is found in more than 95% of traits, regardless of the type of trait, but that the magnitude of the effect in relation to the additive effect is much greater in life-history than in morphological traits. Epistatic effects were detected more often in life-history than in morphological traits (79% and 67%, respectively). We also test for a difference in the magnitude of the effects by comparing the ratio of the nonadditive components separately to the additive component. For both dominance and epistatic components, the ratio of the nonadditive component to additive effects in life history traits is approximately twice as large as that for morphological traits. PMID- 17133856 TI - Protected polymorphism and evolutionary stability in pleiotropic models with trait-specific dominance. AB - When alleles have pleiotropic effects on a number of quantitative traits, the degree of dominance between a pair of alleles can be different for each trait. Such trait-specific dominance has been studied previously in models for the maintenance of genetic variation by antagonistic effects of an allele on two fitness components. By generalizing these models to an arbitrary number of fitness components or other phenotypic traits with different degrees of dominance, I show that genetic polymorphism is generally impossible without antagonistic fitness effects of different traits and without trait-specific dominance. I also investigate dominance and pleiotropy from a more long-term evolutionary perspective, allowing for the study of general ecological scenarios, and I discuss the effects of trait-specific dominance on evolutionary stability criteria. When selection is mainly directional and only trait-specific dominance and antagonism cause the emergence of polymorphism, then these polymorphisms can be overtaken by single mutants again, such that they are probably short-lived on an evolutionary time scale. Near evolutionarily singular points where directional selection is absent, trait-specific dominance and overdominance facilitate the emergence of polymorphism and cause evolutionary divergence in some cases. An important outcome of these models is that trait-specific dominance allows for the emergence of genetic polymorphisms without a selective disadvantage for heterozygotes. This removes the scope for the evolution of assortative mate choice and affects dominance modification. Sympatric speciation by disruptive ecological selection requires this heterozygote disadvantage in order to evolve, and therefore it becomes less plausible if the emergence of genetic polymorphism usually occurs via trait-specific dominance and antagonistic effects. PMID- 17133857 TI - When Rensch meets Bergmann: does sexual size dimorphism change systematically with latitude? AB - Bergmann's and Rensch's rules describe common large-scale patterns of body size variation, but their underlying causes remain elusive. Bergmann's rule states that organisms are larger at higher latitudes (or in colder climates). Rensch's rule states that male body size varies (or evolutionarily diverges) more than female body size among species, resulting in slopes greater than one when male size is regressed on female size. We use published studies of sex-specific latitudinal body size clines in vertebrates and invertebrates to investigate patterns equivalent to Rensch's rule among populations within species and to evaluate their possible relation to Bergmann's rule. Consistent with previous studies, we found a continuum of Bergmann (larger at higher latitudes: 58 species) and converse Bergmann body size clines (larger at lower latitudes: 40 species). Ignoring latitude, male size was more variable than female size in only 55 of 98 species, suggesting that intraspecific variation in sexual size dimorphism does not generally conform to Rensch's rule. In contrast, in a significant majority of species (66 of 98) male latitudinal body size clines were steeper than those of females. This pattern is consistent with a latitudinal version of Rensch's rule, and suggests that some factor that varies systematically with latitude is responsible for producing Rensch's rule among populations within species. Identifying the underlying mechanisms will require studies quantifying latitudinal variation in sex-specific natural and sexual selection on body size. PMID- 17133858 TI - Sex ratios when helpers stay at the nest. AB - This study investigates the evolution of the sex ratio (parental investment in sons) when breeding adults are supported by help provided by nonbreeding individuals of one sex. The study also assumes that the helping sex remains on its natal site to compete for the opportunity to breed, whereas the nonhelping sex disperses. Two kin-selection models are presented, both of which incorporate the age structure found in many natural populations where such helping occurs. The first model assumes that helpers increase the survival of their parents. The second model assumes that helpers are indiscriminant: a helper chooses to increase the survival of a random pair of adults breeding on its natal patch. In both models, sex ratios are not always biased toward the sex that provides the most help. When helpers do not discriminate (second model), the direction of sex ratio bias is determined solely by the size of the benefit of helping behavior. When this benefit is small, sex-ratio evolution is primarily influenced by local resource competition and sex ratios are biased toward the nonhelping (dispersive) sex. If the benefit of help is large enough, the effect of local resource competition is reduced and sex-ratio bias favors the helpful sex. When helpers help only their parents, the same qualitative relationship exists between the direction of sex-ratio bias and the benefit of helping. In this case, however, the direction of sex-ratio bias is also influenced by the size of the social group, mortality, and which individual (mother or father) controls the sex ratio. This study also investigates a sex-ratio conflict that exists between mates. Helping behavior of nonbreeders can act to alleviate the disparities between the optimal sex ratio from the perspective of a mother and that from the perspective of a father. This consequence of helping has not been previously recognized. PMID- 17133859 TI - Lower group productivity under kin-selected reproductive altruism. AB - Hamilton's rule provides the foundation for understanding the genetic evolution of social behavior, showing that altruism is favored by increased relatedness and increased productivity of altruists. But how likely is it that a new altruistic mutation will satisfy Hamilton's rule by increasing the reproductive efficiency of the group? Altruism per se does not improve efficiency, and hence we would not expect a typical altruistic mutation to increase the mean productivity of the population. We examined the conditions under which a mutation causing reproductive altruism can spread when it does not increase productivity. We considered a population divided into temporary groups of genetically similar individuals (typically family groups). We show that the spread of altruism requires a pleiotropic link between altruism and enhanced productivity in diploid organisms, but not in haplodiploid organisms such as Hymenoptera. This result provides a novel biological understanding of the barrier to the spread of reproductive altruism in diploids. In haplodiploid organisms, altruism within families that lowers productivity may spread, provided daughters sacrifice their own reproduction to raise full-sisters. We verified our results using three single-locus genetic models that explore a range of the possible reproductive costs of helping. The advantage of female-to-female altruism in haplodiploids is a well-known prediction of Hamilton's rule, but its importance in relaxing the linkage between altruism and efficiency has not been explored. We discuss the possible role of such unproductive altruism in the origins of sociality. We also note that each model predicts a large region of parameter space were polymorphism between altruism and selfishness is maintained, a pattern independent of dominance. PMID- 17133860 TI - Functional origins of fitness effect-sizes of compensatory mutations in the DNA bacteriophage phiX174. AB - Epistasis is an important and poorly understood aspect of mutations and strongly influences the evolutionary impact of genetic variation on adaptation and fitness. Although recent studies have begun to characterize the distribution of epistatic effects between mutations affecting fitness, there is currently a lack of empirical information on the underlying biological causes of these epistatic interactions. What are the functional constraints that determine the effectiveness of a compensatory mutation at restoring fitness? We have measured the effect-sizes of 52 compensatory mutations affecting nine different deleterious mutations in the major capsid and spike proteins of the DNA bacteriophage phiX174. On average, an experimentally detectable compensatory mutation recovers about two-thirds of the fitness cost of the preceding deleterious mutation. Variation in fitness effect-sizes is only weakly associated with measures of the distance separating the deleterious and compensatory mutations in the amino acid sequence or the folded protein structure. However, there is a strong association of fitness effect-size with the correlation in the effects of the mutations on the biochemical properties of amino acids. A compensatory mutation has the largest effect-size, on average, when both the compensatory and deleterious mutations have radical effects on the overall biochemical make-up of the amino acids. By examining the relative contributions of specific biochemical properties to variation in fitness effect-size, we find that the area and charge of amino acids have a major influence, which suggests that the complexity of the amino acid phenotype is simplified by selection into a reduced number of phenotypic components. PMID- 17133861 TI - Effects of competition on the fitness of wild and crop-wild hybrid sunflower from a diversity of wild populations and crop lines. AB - Gene flow between crop fields and wild populations often results in hybrids with reduced fitness compared to their wild counterparts due to characteristics imparted by the crop genome. But the specifics of the evolutionary outcome of crop-wild gene flow may depend on context, varying due to local environmental conditions and genetic variation within and among wild populations and among crop lines. To evaluate context-dependence of fitness of F1 hybrids, sunflower crop lines were crossed with nine wild populations from across the northern United States. These crop-wild hybrids and their wild counterparts were grown under agricultural conditions in the field with and without wheat competition. Hybrids were far less fecund than wild plants, yet more likely to survive to reproduce. There was considerable variability among wild populations for fecundity and the specific crop line used to generate the crop-wild hybrid significantly affected fecundity. The fitness deficit suffered by crop-wild hybrids varied by population, as did the rankings of the crop-wild hybrids from three different crop lines. Wheat competition decreased fecundity and survival considerably and hampered seed production of wild plants more than that of hybrids. Genotype x environment interactions indicated that the response of fitness to competition differed by population. Consequently, the fitness of hybrids relative to wild plants varied considerably among wild populations and was not consistent across environments. Notably, relative fitness of hybrids was greater under competitive conditions. This research is the first study of its kind to demonstrate that the consequences of crop-wild gene flow are context dependent and contingent on the genetics of the specific wild populations and the local biotic and abiotic conditions. PMID- 17133862 TI - Ovule discounting in an outcrossing, cryptically dioecious tree. AB - Ovule discounting denotes the reduction in the number of ovules available for cross-fertilization due to the interference of inferior pollen. Traditionally, ovule discounting has been discussed solely from the perspective of compromised outcrossing opportunities as a result of selfing, but the principle is more general. Here, we extend its applicability beyond the simple contrast between selfing and outcrossing by showing that, in the cryptically dioecious tree species Fraxinus ornus, ovule discounting through frequent outcrossing with inferior fathers also constitutes a substantial cost of mating. In F. ornus, hermaphrodites produce pollen capable of siring offspring, but these offspring are less viable than those sired by males and are inferred to produce few, if any, surviving progeny. In this paper, we used microsatellite markers to analyze the mating system and paternity in a wild population of F. ornus. We found that the effective number of sires per mother was low (N(ep) = 2.93 to 4.95), and that paternity was correlated among progeny sampled from the same mother, but not among progeny sampled from neighboring mothers. Despite the existence of a local spatial genetic structure (up to 30 m), we found no evidence of biparental inbreeding. There was negligible selfing by hermaphrodites, but they sired approximately one fourth of the seeds produced by other hermaphrodites. Given that these progeny are not inferred to reach reproductive maturity, this constitutes a substantial cost of ovule discounting in the broad sense. We discuss the possible reasons for why hermaphrodites invest resources into inferior pollen. PMID- 17133863 TI - Mechanisms underlying the sperm quality advantage in Drosophila melanogaster. AB - Contrary to early predictions of sperm competition theory, postcopulatory sexual selection favoring increased investment per sperm (e.g., sperm size, sperm quality) has been demonstrated in numerous organisms. We empirically demonstrate for Drosophila melanogaster that both sperm quality and sperm quantity independently contribute to competitive male fertilization success. In addition to these independent effects, there was a significant interaction between sperm quality and quantity that suggests an internal positive reinforcement on selection for sperm quality, with selection predicted to intensify as investment per sperm increases and the number of sperm competing declines. The mechanism underlying the sperm quality advantage is elucidated through examination of the relationship between female sperm-storage organ morphology and the differential organization of different length sperm within the organ. Our results exemplify that primary sex cells can bear secondary sexual straits. PMID- 17133864 TI - Within-population variation in cytoplasmic genes affects female life span and aging in Drosophila melanogaster. AB - It has been suggested that mitochondrial DNA (mtDNA) may play an important role in aging. Yet, few empirical studies have tested this hypothesis, partly because the degree of sequence polymorphism in mtDNA is assumed to be low. However, low sequence variation may not necessarily translate into low phenotypic variation. Here, we report an experiment that tests whether there is within-population variation in cytoplasmic genes for female longevity and senescence. To achieve this, we randomly selected 25 "mitochondrial founders" from a single, panmictic population of Drosophila melanogaster and used these founders to generate distinct "mt" lines in which we controlled for the nuclear background by successive backcrossing. Potential confounding effects of cytoplasmically transmitted bacteria were eliminated by tetracycline treatment. The mt lines were then assayed for differences in longevity, Gompertz intercept (frailty), and demographic rate of change in mortality with age (rate-of-senescence) in females. We found significant cytoplasmic effects on all three variables. This provides evidence that genetic variation in cytoplasmic genes, presumably mtDNA, contributes to variation in female mortality and aging. PMID- 17133865 TI - Parents benefit from eating offspring: density-dependent egg survivorship compensates for filial cannibalism. AB - Why should animals knowingly consume their own young? It is difficult to imagine many circumstances in which eating one's own young (i.e., filial cannibalism) actually increases an individual's fitness; however, filial cannibalism commonly co-occurs with parental care in fishes. The evolutionary significance of filial cannibalism remains unclear. The most commonly accepted explanation is that filial cannibalism is a mechanism by which caring males gain energy or nutrients that they reinvest into future reproduction, thereby increasing net reproductive success. There is mixed support for this hypothesis and, at best, it can only explain filial cannibalism in some species. A recent alternative hypothesis suggests that filial cannibalism improves the survivorship of remaining eggs by increasing oxygen availability, and thus increases current reproductive success. This theory has received little attention as of yet. We evaluated the hypothesis of oxygen-mediated filial cannibalism in the sand goby by examining the effect of oxygen and egg density on the occurrence of filial cannibalism, evaluating the effects of partial clutch cannibalism on the survivorship of remaining eggs, and comparing potential costs and benefits of filial cannibalism related to the net number of eggs surviving. Indeed, we found that oxygen level and egg density affected the occurrence of cannibalism and that simulated partial clutch cannibalism improved survivorship of the remaining eggs. Additionally, because increased egg survivorship, stemming from partial egg removal, compensated for the cost of cannibalism (i.e., number of eggs removed) at a range of cannibalism levels, filial cannibalism potentially results in no net losses in reproductive success. However, oxygen did not affect egg survivorship. Thus, we suggest a more general hypothesis of filial cannibalism mediated by density-dependent egg survivorship. PMID- 17133866 TI - Micro- and macroevolutionary decoupling of cichlid jaws: a test of Liem's key innovation hypothesis. AB - The extent to which elements of functional systems can change independently (modularity) likely influences the diversification of lineages. Major innovations in organismal design, like the pharyngeal jaw in cichlid fishes, may be key to a group's success when they relax constraints on diversification by increasing phenotypic modularity. In cichlid fishes, pharyngeal jaw modifications that enhanced the ability to breakdown prey may have freed their oral jaws from serving their ancestral dual role as a site of both prey capture and prey processing. This functional decoupling that allowed the oral jaws to become devoted solely to prey capture has been hypothesized to have permitted the two sets of cichlid jaws to evolve independently. We tested the hypothesis that oral and pharyngeal jaw mechanics are evolutionarily decoupled both within and among Neotropical Heroine cichlids. In the trophically polymorphic species Herichthys minckleyi, molariforms that exhibit enlarged molarlike pharyngeal jaw teeth were found to have approximately 400% greater lower jaw mass compared to H. minckleyi with the alternative papilliform pharyngeal morphology. However, oral jaw gape, lower jaw velocity ratios, anterior jaw linkage mechanics, and jaw protrusion did not differ between the morphotypes. In 40 other Heroine species, there was a weak correlation between oral jaw mechanics and pharyngeal jaw mass when phylogenetic history was ignored. Yet, after expansion of the cytochrome b phylogeny for Heroines, change in oral jaw mechanics was found to be independent of evolutionary change in pharyngeal jaw mass based on independent contrasts. Evolutionary decoupling of oral and pharyngeal jaw mechanics has likely played a critical role in the unparalleled trophic diversification of cichlid fishes. PMID- 17133867 TI - Triturus newts defy the running-swimming dilemma. AB - Conflicts between structural requirements for carrying out different ecologically relevant functions may result in a compromise phenotype that maximizes neither function. Identifying and evaluating functional trade-offs may therefore aid in understanding the evolution of organismal performance. We examined the possibility of an evolutionary trade-off between aquatic and terrestrial locomotion in females of European species of the newt genus Triturus. Biomechanical models suggest a conflict between the requirements for aquatic and terrestrial locomotion. For instance, having an elongate, slender body, a large tail, and reduced limbs should benefit undulatory swimming, but at the cost of reduced running capacity. To test the prediction of an evolutionary trade-off between swimming and running capacity, we investigated relationships between size corrected morphology and maximum locomotor performance in females of ten species of newts. Phylogenetic comparative analyses revealed that an evolutionary trend of body elongation (increasing axilla-groin distance) is associated with a reduction in head width and forelimb length. Body elongation resulted in reduced maximum running speed, but, surprisingly, also led to a reduction in swimming speed. The evolution of longer tails was associated with an increase in maximal swimming speed. We found no evidence for an evolutionary trade-off between aquatic and terrestrial locomotor performance, probably because of the unexpected negative effect of body elongation on swimming speed. We conclude that the idea of a design conflict between aquatic and terrestrial locomotion, mediated through antagonistic effects of body elongation, does not apply to our model system. PMID- 17133868 TI - Faster lizards sire more offspring: sexual selection on whole-animal performance. AB - Sexual selection operates by acting on variation in mating success. However, since selection acts on whole-organism manifestations (i.e., performance) of underlying morphological traits, tests for phenotypic effects of sexual selection should consider whole-animal performance as a substrate for sexual selection. Previous studies have revealed positive relationships between performance and survival, that is, natural selection, but none have explicitly tested whether performance may influence reproductive success (through more matings), that is, sexual selection. Performance predicts dominance in some species, implying the effects of sexual selection, but how it does so has not been established, nor is it certain whether performance might be a by-product of selection for something else, for example, elevated circulating testosterone levels. We investigated the potential for sexual selection on sprint speed performance in collared lizards (Crotaphytus collaris), considering the potential mediating effects of circulating hormone levels. Among territorial, adult male collared lizards, only sprint speed significantly predicted territory area and number of offspring sired as determined by genetic paternity analysis. Body size, head size, and hind limb length had no effect. Neither plasma testosterone levels nor corticosterone levels correlated with sprint speed, territory area, or number of offspring sired. Thus, our results provide a direct link between whole-animal performance and reproductive success, suggesting that intrasexual selection can act directly on sprint speed performance and drive the evolution of underlying morphological traits. PMID- 17133869 TI - No seasonal sex-ratio shift despite sex-specific fitness returns of hatching date in a lizard with genotypic sex determination. AB - Sex allocation theory predicts that mothers should adjust their sex-specific reproductive investment in relation to the predicted fitness returns from sons versus daughters. Sex allocation theory has proved to be successful in some invertebrate taxa but data on vertebrates often fail to show the predicted shift in sex ratio or sex-specific resource investment. This is likely to be partly explained by simplistic assumptions of vertebrate life-history and mechanistic constraints, but also because the fundamental assumption of sex-specific fitness return on investment is rarely supported by empirical data. In short-lived species, the time of hatching or parturition can have a strong impact on the age and size at maturity. Thus, if selection favors adult sexual-size dimorphism, females can maximize their fitness by adjusting offspring sex over the reproductive season. We show that in mallee dragons, Ctenophorus fordi, date of hatching is positively related to female reproductive output but has little, if any, effect on male reproductive success, suggesting selection for a seasonal shift in offspring sex ratio. We used a combination of field and laboratory data collected over two years to test if female dragons adjust their sex allocation over the season to ensure an adaptive match between time of hatching and offspring sex. Contrary to our predictions, we found no effect of laying date on sex ratio, nor did we find any evidence for within-female between-clutch sex ratio adjustment. Furthermore, there was no differential resource investment into male and female offspring within or between clutches and sex ratios did not correlate with female condition or any partner traits. Consequently, despite evidence for selection for a seasonal sex-ratio shift, female mallee dragons do not seem to exercise any control over sex determination. The results are discussed in relation to potential constraints on sex-ratio adjustment, alternative selection pressures, and the evolution of temperature-dependent sex determination. PMID- 17133870 TI - The quick and the fast: the evolution of acceleration capacity in Anolis lizards. AB - Although of prime ecological relevance, acceleration capacity is a poorly understood locomotor performance trait in terrestrial vertebrates. No empirical data exist on which design characteristics determine acceleration capacity among species and whether these design traits influence other aspects of locomotor performance. In this study we explore how acceleration capacity and sprint speed have evolved in Anolis lizards. We investigate whether the same or different morphological traits (i.e., limb dimensions and muscle mass) correlate with both locomotor traits. Within our sample of Anolis lizards, relative sprint speed and acceleration capacity coevolved. However, whereas the variation in relative acceleration capacity is primarily explained by the variation in relative knee extensor muscle mass, the variation in relative sprint speed is correlated to the variation in relative femur, tibia, and metatarsus length as well as knee extensor muscle mass. The fact that the design features required to excel in either performance trait partly overlap might explain the positive correlation between the variation in relative sprint speed and acceleration capacity. Furthermore, our data show how similar levels of sprint performance can be achieved through different morphological traits (limb segment lengths and muscle mass) suggesting that redundant mapping has potentially played a role in mitigating trade-offs. PMID- 17133871 TI - Calling is an honest indicator of paternal genetic quality in poison frogs. AB - Several competing hypotheses have been put forward to explain why females of many species mate preferentially with males possessing the most conspicuous signals (e.g., ornaments, displays, or songs). We performed a laboratory experiment using two species of poison frogs, Dendrobates leucomelas and Epipedobates tricolor, to test the hypothesis that male calling performance is an honest indicator of parental quality. Our analyses are based on data from behavioral observations of mating activities of captive-reared individuals (and their offspring) that were housed in terraria for four consecutive breeding seasons. Male mating success increased with male calling rate and chirp duration in both species, suggesting that females preferred males with more elaborate calls. Because calling performance improved with age in D. leucomelas, female poison frogs that prefer males with more elaborate calls in the wild may end up mating with older males that have already proven their ability to survive. Females that mated with good callers obtained higher quality offspring. Eggs fertilized by males with high calling rates and long chirp durations had higher hatching success and produced tadpoles that were more likely to metamorphose into surviving frogs. As a consequence, females that mated with males with high calling performance obtained more surviving offspring per egg, compared to females that mated with poor callers. Collectively, our findings comply with the notion that female poison frogs prefer to mate with good callers because calling performance is a reliable predictor of offspring quality. The possible influence of maternal allocation and reasons for the strong effect size compared to previous studies are discussed. PMID- 17133872 TI - The role of body size in host specificity: reciprocal transfer experiments with feather lice. AB - Although most parasites show at least some degree of host specificity, factors governing the evolution of specificity remain poorly understood. Many different groups of host-specific parasites show a striking correlation between parasite and host body size, suggesting that size reinforces specificity. We tested this hypothesis by measuring the relative fitness of host-specific feather lice transferred to pigeons and doves that differ in size by an order of magnitude. To test the general influence of size, we transferred unrelated groups of wing and body lice, which are specialized for different regions of the host. Lice were transferred in both directions, from a large native host species, the rock pigeon (Columba livia), to several progressively smaller hosts, and from a small native host species, the common ground dove (Columbina passerina), to several larger hosts. We measured the relative fitness (population size) of lice transferred to these novel host species after two louse generations. Neither wing lice nor body lice could survive on novel host species that were smaller in size than the native host. However, when host defense (preening behavior) was blocked, both groups survived and reproduced on all novel hosts tested. Thus, host defense interacted with host size to govern the ability of lice to establish on small hosts. Neither wing lice nor body lice could survive on larger hosts, even when preening was blocked. In summary, host size influenced the fitness of both types of feather lice, but through different mechanisms, depending on the direction of the transfer. Our results indicate that host switching is most likely between hosts of similar body size. This finding has important implications for studies of host-parasite coevolution at both the micro- and macroevolutionary scales. PMID- 17133873 TI - Live fast, die young: trade-offs between fitness components and sexually antagonistic selection on weaponry in Soay sheep. AB - Males are predicted to compete for reproductive opportunities, with sexual selection driving the evolution of large body size and weaponry through the advantage they confer for access to females. Few studies have explored potential trade-offs of investment in secondary sexual traits between different components of fitness or tested for sexually antagonistic selection pressures. These factors may provide explanations for observed polymorphisms in both form and quality of secondary sexual traits. We report here an analysis of selection on horn phenotype in a feral population of Soay sheep (Ovis aries) on the island of Hirta, St. Kilda, Scotland. Soay sheep display a phenotypic polymorphism for horn type with males growing either normal or reduced (scurred) horns, and females growing either normal, scurred, or no (polled) horns; further variation in size exists within horn morphs. We show that horn phenotype and the size of the trait displayed is subject to different selection pressures in males and females, generating sexually antagonistic selection. Furthermore, there was evidence of a trade-off between breeding success and longevity in normal-horned males, with both the normal horn type and larger horn size being associated with greater annual breeding success but reduced longevity. Therefore, selection through lifetime breeding success was not found to act upon horn phenotype in males. In females, a negative association of annual breeding success within the normal horned phenotype did not result in a significant difference in lifetime fitness when compared to scurred individuals, as no significant difference in longevity was found. However, increased horn size within this group was negatively associated with breeding success and longevity. Females without horns (polled) suffered reduced longevity and thus reduced lifetime breeding success relative the other horn morphs. Our results therefore suggest that trade-offs between different components of fitness and antagonistic selection between the sexes may maintain genetic variation for secondary sexual traits within a population. PMID- 17133874 TI - The promise of real-time continuous glucose sensors. PMID- 17133875 TI - New Utah program helps curb runaway costs associated with chronic pain. AB - Take steps to address chronic pain now. Why? Because it's costing the United States 120 billion dollars per year, and much of this expense is due to patients bouncing from clinician to clinician without ever finding relief. Some healthcare organizations and individual practitioners successfully address the problem by taking a multidisciplinary approach to care and equipping patients with strategies to help them take charge. PMID- 17133876 TI - Unprecedented guidelines focus on care of patients with advanced HF. AB - Setting the standard for patients with advanced HF. Although heart transplants are performed in much the same way all over the world, standards vary in regard to listing and selecting patients for transplants. This encourages patients and families to search for programs that offer the most advantageous criteria. However, an international panel of experts has now agreed on criteria that they hope will not only standardize the care of patients with advanced HF, but also create a fairer and more equitable system for listing and selecting transplant candidates. PMID- 17133877 TI - Home care may lead to cost-efficient management of frail elderly patients. AB - House calls make a return in new model of care for frail elderly. Care for such patients is often so fragmented and expensive that the pioneers behind a Woodland Hills, CA, company believe that they can do a better job at less expense by delivering care in the home and giving patients direct access to their physicians at any time of the day or night. Some commercial health plans have already bought into the approach, and now CMS is taking a close look at the model as part of a three-year demonstration project. PMID- 17133878 TI - When doctors lead contracting efforts, orthopedic implant savings will follow. PMID- 17133880 TI - Prices rise along with business activity in August. But ISM purchasing survey sees price stability in healthcare. PMID- 17133879 TI - Orthopedic prices don't match case volume, survey shows. PMID- 17133881 TI - It's not your ordinary flu. PMID- 17133882 TI - Physician/pharmacist collaboration--regulations on the way! PMID- 17133883 TI - The Iowa Healthcare Collaborative: the power of coming together. PMID- 17133884 TI - Malpractice: the hidden, emotional costs. PMID- 17133885 TI - Type of items and the magnitude of gender differences on the Mental Rotations Test. AB - The present study considered the structure of the drawings used in the Mental Rotations Test (MRT) to examine whether distractors that are either a mirror image or structurally different from the target as well as response alternatives with occluded and nonoccluded parts affect the magnitude of gender differences on this test. One hundred and three women and 100 men undergraduate students were given unlimited time to complete the MRT. A gender by occlusion interaction on correct responses showed that gender differences were larger for occluded than for nonoccluded items. Examination of performance as a function of item placement in the test suggested that it is unlikely that the results could be accounted for by differential practice effects in women and men. Implications of these results for explanations of gender differences on the MRT and for the training of spatial abilities are discussed. PMID- 17133886 TI - Fundamental properties of the N2pc as an index of spatial attention: effects of masking. AB - Masking is an important tool in many paradigms used to study the cognitive architecture. The N2pc is an electrophysiological event-related potential (ERP) that has been used as a tool to study the deployment of visual spatial attention. The aim of this paper was to study the effects of masking on the N2pc. Two stimuli were presented on the screen, one to left and one to right of fixation, and subjects reported the identity of one of them. The targets could be discriminated both by their category (letters vs. digit) and by their colour (pink vs. green). Backward masking was produced by presenting a second pair of bilateral stimuli after the offset of the first pair. The second pair of stimuli consisted of characters of the same colour and category as in the first pair. Forward masking was produced by using the very same stimuli as in the backward masking condition, but by instructing subjects to report the second stimulus. The forward mask trials had longer response times compared to no-mask trials, and backward mask trials had even longer response times, and also a higher error rate. Although the different masking procedures lead to clear behavioural effects, the N2pc was not affected, suggesting that the deployment of visual spatial attention, per se, was not affected by pattern masking. A sustained posterior contralateral negativity (SPCN) following the N2pc was also found (300 ms post-target, and beyond), and the amplitude of the SPCN was strongly modulated by the number of presented stimuli and the duration of the SPCN was positively correlated with RT in the behavioural task. We hypothesize that the SPCN reflects neural activity associated with the passage of information through visual short term memory. PMID- 17133887 TI - Parametric exploration of the Simon effect across visual space. AB - The Simon effect refers to the performance advantage for responding to the nonspatial identity of the target when the target's irrelevant location corresponds with the relative location of the response. The present study is a parametric examination of the magnitude of the Simon effect across visual space. Response keys were arranged along vertical, horizontal, and two diagonal axes, and stimuli were arranged in two concentric circles (near and far from fixation) along the same axes. The results show that the Simon effect is of similar magnitude regardless of stimulus-response axis. In contrast to findings from stimulus-response compatibility paradigms, there was no evidence in this study for the presence of an orthogonal compatibility effect or left-right prevalence effect, suggesting that these effects only arise when response location is relevant. The results demonstrate the robust generalizability of the Simon effect under different spatial conditions and thus broaden the relevance of the Simon effect to a variety of applications. PMID- 17133889 TI - Basic processes in reading: can functional phonological recoding be blocked? AB - A widely held view is that phonological processing is always involved in lexical access from print, and is automatic in that it cannot be prevented. This claim was assessed in the context of a priming paradigm. In Experiment 1, repetition priming was observed for both pseudohomophone-word pairs (e.g., brane-brain) and morphologically related word pairs (e.g., marked-mark) in the context of lexical decision. In Experiment 2, subjects searched the prime for the presence of a target letter and then made a lexical decision to a subsequent letter string. Phonological priming from a pseudohomophone was eliminated following letter search of the prime, whereas morphological priming persisted. These results are inconsistent with the claim that a) lexical access from print requires preliminary phonological processing, and b) functional phonological processing cannot be blocked. They are, however, consistent with the conclusion that, for intact skilled readers, lexical access can be accomplished on the basis of orthographic processing alone. These results join a growing body of evidence supporting the claim that there exist numerous points in visual word recognition at which processing can be stopped. PMID- 17133890 TI - The orthographic neighbourhood frequency effect in French: a letter-case manipulation study. AB - The neighbourhood frequency effect was investigated by a letter-case manipulation of French words presented in the go/no-go lexical decision task. Accented words were presented both in lower-case letters with written accents and upper-case letters with no written accents, two usual typographies in French. This procedure addressed the stimulus-matching problem by using intra-word comparisons as some words have different orthographic neighbourhoods in the two cases. Neighbourhood frequency was varied across case change for half of the words while it was held constant for the other half. The results showed an interaction between letter case and neighbourhood constancy, so the inhibitory neighbourhood frequency effect could not be attributed to interstimulus comparisons or to typographical change. Implications are discussed in current models of visual word recognition. PMID- 17133891 TI - Growth points in research on memory and hippocampus. AB - We present an overview of two of our on-going projects relating processes in the hippocampus to memory. We are trying to understand why retrograde amnesia occurs after damage to the hippocampus. Our experiments establish the generality of several new retrograde amnesia phenomena that are at odds with the consensus view of the role of the hippocampus in memory. We show in many memory tasks that complete damage to the hippocampus produces retrograde amnesia that is equivalent for recent and remote memories. Retrograde amnesia affects a much wider range of memory tasks than anterograde amnesia. Normal hippocampal processes can interfere with retention of a long-term memory stored outside the hippocampus. We conclude that the hippocampus competes with nonhippocampal systems during memory encoding and retrieval. Finally, we outline a project to understand and manipulate adult hippocampal neurogenesis in order to repair damaged hippocampal circuitry to recover lost cognitive functions. PMID- 17133892 TI - Autonomy, paternalism, and the doctor's duty of care. PMID- 17133893 TI - Enhancing meaning in palliative care practice: a meaning-centered intervention to promote job satisfaction. AB - OBJECTIVES: This article introduces a new meaning-centered psycho-educational group intervention, called Enhancing meaning in palliative care nursing, designed to support nurses providing palliative care. This intervention aims at increasing job satisfaction and quality of life, as well as preventing burnout in this particular population. THEORETICAL FRAMEWORKS: Its format and content are founded on the meaning-centered psychotherapy approach developed for terminally ill cancer patients (Breitbart, 2001; Greenstein & Breitbart, 2000). Frankl's existential therapeutic approach, called logotherapy, serves as the underlying theoretical framework to this intervention. DEVELOPMENT: Following the presentation of the context and the development of the intervention, its content is described. CONCLUSION: A brief description of the ongoing randomized controlled trial testing the intervention is then provided. Finally, the way in which this intervention could contribute to nurses' quality of life and suggestions for future developments are briefly discussed. PMID- 17133894 TI - Religion and spirituality among patients with localized prostate cancer. AB - OBJECTIVE: To examine: (1) daily religious and spiritual experiences among localized prostate cancer patients as compared to a national age and race-matched male sample; (2) cognitive-affective and clinical predictors of prostate cancer diagnosis-related increases in religiosity and spirituality; (3) short-term impact of daily religious and spiritual experiences on cancer recurrence worry. METHODS: Analyses of data from a longitudinal questionnaire study among patients (N = 254) diagnosed with localized prostate cancer and data from a random sample (N = 238) of respondents to the national General Social Survey. RESULTS: Compared to the national sample, prostate cancer patients reported higher levels of daily spiritual experiences. Patients with higher worry about prostate cancer and elevated levels of prostate-related symptoms around diagnosis were more likely to report a diagnosis-related increase in religiosity and spirituality. Positive benefits (reduced recurrence worry) of religious coping/practices were restricted to those patients with higher versus lower level of postdiagnosis increase in religiosity; patients not reporting postdiagnosis increases in religion who are not engaging in religious coping/practice adjusted equally well. Results suggest that the development of religious/spiritual interventions is premature. SIGNIFICANCE OF RESULTS: This is the first prospective study to report on the prevalence and influence of daily spiritual and religious experiences among prostate cancer patients. PMID- 17133895 TI - The relation between the experience of time and psychological distress in patients with hematological malignancies. AB - OBJECTIVE: The experience of time is strongly related to our momentary mood states. Patients with a life-threatening illness experience an extreme change in mood and suffer from psychological distress that can develop into clinically relevant psychiatric disorders, like anxiety and depression. The aim of this study was to investigate the associations among the subjective perception of time, psychological distress, and quality of life in patients with hematological malignancies. METHODS: Eighty-eight inpatients with hematological malignancies rated how fast time passes subjectively on a visual analog scale and prospectively estimated a time span of 13 min. The Hospital Anxiety and Depression Scale (HADS) self-report measures of health-related quality of life (FACT-G) and spiritual well-being (FACIT-Sp) were employed to assess psychological distress and quality of life. RESULTS: Those patients who reported a lower quality of life, less spiritual well-being, and more anxiety experienced a slower passage of subjective time and overestimated the 13-min time interval. SIGNIFICANCE OF RESULTS: Our interpretation of the results is that patients with a life-threatening illness who show symptoms of psychological distress draw attention away from meaningful thoughts and actions and, thus, experience time as passing more slowly. An altered sense of time can be a sign of mental suffering, which should be addressed within psycho-oncological interventions. As this is the first study to demonstrate this relation in cancer patients, further research is needed to investigate the experience of time and its relation to meaning as an issue in clinical diagnostics. PMID- 17133896 TI - A qualitative study of the trauma and posttraumatic growth of multiple myeloma patients treated with peripheral blood stem cell transplant. AB - OBJECTIVE: The study was conducted to understand the emotional impact of multiple myeloma, as well as the impact of its principle treatment, peripheral blood stem cell transplant (PBSCT). The absence of psycho-oncology research literature on this population prompted the need for a hypothesis-generating investigation. Thus, a qualitative design was used to construct a theoretical model of the trauma relating to diagnosis and treatment of myeloma. The study also incorporates the important period of reflection and growth following treatment. METHODS: The sample consisted of 3 women and 3 men treated for myeloma at a New York City-based cancer treatment center. Data from individual interviews were audiotaped and transcribed. After extensive review, the data were categorized into groups of repeating ideas, themes and broad theoretical constructs. RESULTS: A five-construct model emerged from the data analysis that integrated a model of trauma and growth presented in earlier work (Auerbach et al., 2006). These constructs roughly correspond with stages of illness, but do not necessarily imply a linear process, as suggested by stage models. The first construct is diagnosis. Patients receive the news that they have multiple myeloma. Initial reactions are discussed and a treatment plan takes form. In the second construct, treatment, patients highlight the physical and emotional hurdles confronted throughout treatment. The third construct, network of safety, presents social factors that play a role in comforting patients throughout illness. Patients recognize the importance of a strong support system during their experiences. In the fourth construct, recuperation, physical energy is regained after an arduous recovery period. This contributes to higher spirits and a motivation to reengage with life. The fifth construct is reflection and new existence. Patients strive to balance a new reality that relapse and death are inevitable, along with their need to live a meaningful life. Many do not yet appreciate how their disease has impacted them, but describe how their interpersonal lives and perceptions have changed, both positively and negatively. SIGNIFICANCE OF RESULTS: Limitations of the study, future directions for research and clinical implications are discussed. PMID- 17133897 TI - Attitudes toward disease and prognosis disclosure and decision making for terminally ill patients in Japan, based on a nationwide random sampling survey of the general population and medical practitioners. AB - OBJECTIVE: Employing a nationwide cross-sectional survey, we investigated the Japanese general population's attitudes toward disease and prognosis disclosure and related factors. Furthermore, we investigated Japanese medical practitioners' attitudes toward disease and prognosis disclosure for patients and decision making. METHODS: A nationwide anonymous questionnaire survey was conducted. A total of 5000 individuals were randomly sampled from the general population and 3104 physicians and 6059 nurses were randomly sampled in Japan. RESULTS: Finally, 2422 people from the general population (response rate, 48%), 1577 physicians (51%), and 3361 nurses (56%) returned questionnaires. Among the general population, 73% of participants answered that they "want to know" about their disease and prognosis when in an incurable disease state. Ninety percent desired direct disclosure and 8% disclosure through their family. However, few medical practitioners answered "patient himself" (physician 3%, nurses 4%) as the person whom they would primarily notify about the disease and prognosis when in charge of a patient with an incurable disease. On the other hand, physicians answered "family" most frequently (59%), whereas nurses most commonly responded, "depends on patient's condition" (63%). SIGNIFICANCE OF RESEARCH: Several detailed analyses of factors associated with prognosis disclosure were conducted. Japanese physicians need to carefully communicate with the patients individually about whether direct disclosure or disclosure primarily to the family is preferred. PMID- 17133898 TI - Requests for euthanasia and physician-assisted suicide and the availability and application of palliative options. AB - OBJECTIVE: This study investigated the palliative options available when a patient requested euthanasia or physician-assisted suicide (EAS), the extent to which the options were applied, and changes in the patient's wishes. METHODS: In an observational study, 3614 general practitioners (GPs) filled in a questionnaire and described their most recent request for EAS (if any) (n = 1,681). RESULTS: Palliative options were still available in 25% of cases. In these cases options were applied in 63%; in 46% of these cases patients withdrew their request. Medication other than antibiotics, which was most frequently mentioned as a palliative option (67%), and applied most frequently (79%), together with radiotherapy, most frequently resulted in patients withdrawing their request. SIGNIFICANCE OF RESULTS: GPs include the availability of palliative options in their decision making when considering EAS. The fact that not all options are applied or, if applied, the patient persists in the request is related to autonomy of the patient, the burden on the patient, and medical futility of the option. PMID- 17133899 TI - Perspectives on spirituality at the end of life: a meta-summary. AB - OBJECTIVE: A meta-summary of the qualitative literature on spiritual perspectives of adults who are at the end of life was undertaken to summarily analyze the research to date and identify areas for future research on the relationship of spirituality with physical, functional, and psychosocial outcomes in the health care setting. METHODS: Included were all English language reports from 1966 to the present catalogued in PubMed, Medline, PsycInfo, and CINAHL, identifiable as qualitative investigations of the spiritual perspectives of adults at the end of life. The final sample includes 11 articles, collectively representing data from 217 adults. RESULTS: The preponderance of participants had a diagnosis of cancer; those with HIV/AIDS, cardiovascular disease, and ALS were also represented. Approximately half the studies were conducted in the United States; others were performed in Australia, Finland, Scotland, and Taiwan. Following a process of theme extraction and abstraction, thematic patterns emerged and effect sizes were calculated. A spectrum of spirituality at the end of life encompassing spiritual despair (alienation, loss of self, dissonance), spiritual work (forgiveness, self exploration, search for balance), and spiritual well-being (connection, self actualization, consonance) emerged. SIGNIFICANCE: The findings from this meta summary confirm the fundamental importance of spirituality at the end of life and highlight the shifts in spiritual health that are possible when a terminally ill person is able to do the necessary spiritual work. Existing end-of-life frameworks neglect spiritual work and consequently may be deficient in guiding research. The area of spiritual work is fertile ground for further investigation, especially interventions aimed at improving spiritual health and general quality of life among the dying. PMID- 17133900 TI - Psychological intervention in fluid management. AB - BACKGROUND: Hemodialysis is a palliative treatment for patients with established renal failure (ERF), and volume overload is a common problem for hemodialysis patients with low urinary output. Volume overload is thought to be mostly attributable to interdialytic fluid intake by the patient and is associated with an increased symptom burden and the development of serious medical complications. Repeated episodes of volume overload may adversely affect staff-patient relationships and the perception of care in this patient population. The aim of this case series study was to evaluate the effect and experience of a psychological intervention on interdialytic weight gain in a small group of patients. METHODS: Five patients were treated. The intervention involved using techniques derived from both cognitive behavior therapy and motivational interviewing. The main outcome measures were interdialytic weight gain and patient perception of the intervention. RESULTS: Three of the five patients reduced both mean interdialytic weight gain and the frequency with which they gained in excess of 3% of their dry weight during the intervention phase. The intervention was found to be acceptable to patients. SIGNIFICANCE OF RESULTS: The intervention was effective in helping three of the five patients to reduce both the frequency and the severity of volume overload, and two of these patients maintained this for at least 6 months post intervention. The intervention used actively engaged the patients and appeared to be experienced positively. The methods used to mobilize patient resources and optimize staff-patient relationships as vehicles of change are discussed. Both may have implications for treatment concordance and the perception of care delivered. PMID- 17133901 TI - [Comparison of disk-diffusion method and PCR for detection of methicillin resistance in Staphylococcus aureus strains]. AB - The aim of the study was to compare the disk-diffusion (oxacillin 1 microg, cefoxitin 30 microg) method and PCR for detection of methicillin-resistance in S. aureus. The investigation were carried out on 120 S. aureus strains isolated from clinical materials of patients hospitalized in the University Hospital at the L. Rydygier Collegium Medicum in Bydgoszcz, University of Nicolaus Copernicus in Torun. Of the 120 S. aureus strains tested, 60 (50%) were mecA-positive by PCR. Consistency of results between oxacillin disk-difussion method and PCR amounted 92.5% and cefoxitin disk-diffusion method and PCR--98.3%. The oxacillin disk difussion method falsely identified 3 (2.5%) strains as MSSA (sensitivity 95.0%) and 4 strains as MRSA (specificity 93.3%) in comparison with PCR. The cefoxitin disk-diffusion method falsely identified 2 (1.6%) strains as MSSA (sensitivity 96.7%) and there were no false resistant results (specificity 100%). Our results showed that in disk-diffusion tests, cefoxitin is a better than oxacillin for the identification of MRSA. PMID- 17133902 TI - [Influence of testing conditions on the susceptibility results of Staphylococcus cohnii to beta-lactams]. AB - The high occurence of coagulase-negative staphylococci among bacteria responsible for hospital infections is unquestioned. Studies on the poorly-known novobiocin resistant, coagulase-negative Staphylococcus cohnii were undertaken. The possibilities of optimizing conditions for determination of susceptibility to beta-lactam antibiotics of this species were researched. In the case of S. cohnii the new cefoxitin test for detection of methicillin resistant strains, introduced by the National Reference Centre for Antibiotics in Poland was found as a good and of credible quality. It was also shown, that application in in vitro examination conditions stimulating the mechanisms of resistance to beta-lactam antibiotics, supplies credible results relating to their true susceptibility. The necessity of establishing individual conditions for susceptibility determination in different species of coagulase-negative staphylococci was suggested. PMID- 17133903 TI - [Occurrence of macrolide-lincosamide-streptogramin B resistant isolates of Staphylococcus aureus in clinical specimens in 2003-2004]. AB - The aim of the study was the analysis of frequency of macrolide-lincosamide streptogramin B (MLS(B)) resistance among MSSA (n=1682) and MRSA (n=272) strains which were isolated in 2002-2004 from various clinical materials from patients hospitalized in the University Hospital at the L. Rydygier Collegium Medicum in Bydgoszcz, University of Nicolaus Copernicus in Torun. Susceptibility testing and examination of methicillin-resistant strains were performed by the disc diffusion techniques according to recommendation of NCCLS. Resistance to the MLS(B) antimicrobials agents was higher among MRSA compared to MSSA isolates. The MLS, constitutive phenotype was more prevalent than the MLS(B) inducible phenotype among investigated MRSA (65.4%) and MSSA (7.6%) isolates. Inducible resistance had only 2.5% of the MSSA and 2.6% of the MRSA strains. Moreover in 2004 there were found increasing frequency of inducible MLS(B) resistance from 1.1% to 5.7% and decreasing frequency of constitutive MLS(B) resistance from 9.2% to 4.7% among MSSA strains, in comparison to 2003. The investigated MSSA MLS(B)-, MLS(B)- and MRSAMLS(B)+, MLS(B)- strains were the most frequently isolated from pus (adequately 5.2%, 28.8% and 30.5%, 10.7%) and also from nosopharynx swabs (1.7% MSSA MLS(B)+ and 22.9% MSSA MLS(B)-) and biomaterials (15,1% MRSA MLS(B)+ and 9.6% MRSA MLS(B)-). They mainly came from patients of the outpatient clinic (2,4% MSSA MLS(B)+ and 19.9% MSSA MLS(B)-) and patients treated at the neurosurgical ward (20.6% MRSA MLS(B)+ and 12.1% MRSA MLS(B)-). PMID- 17133904 TI - [Occurrence of adhesin genes in coagulase-negative Staphylococcus aureus strains]. AB - Sixty fenotypically coagulase-negative Staphylococcus aureus strains were screened for the presence of six adhesion genes. The strains were isolated from varied clinical samples and varied patients in 16 medical centers, in majority from the region of Gdansk. Multiplex PCR in two primer sets was used for detection of the following genes: bbp (bone binding protein), cna (collagen binding protein), ebp (elastin binding protein)and fnbB (fibronectin B binding protein), fib (fibrinogen bindng protein) and clfA (clunmping factor A). More than half (57%) of the examined population harbored four genes: fnbB,fib, cna i clfA. All of the strains were found to be clfA positive and 90% of them were positive for fnbB, 90% for fib and 67% for cna. All of these genes were significantly more common in MRSA than in MSSA. The particular genes were occurred in strains derived from varied clinical samples. PMID- 17133905 TI - [Characteristic of Listeria spp. bacteria isolated from food products]. AB - The frequency of occurrance of Listeria strains in different food products was determined. Biochemical characteristic of the isolated strains was achieved in accordance with procedure included in PNEN ISO 11290 standard, genus was determined byApiListeria (bioMeieux) test. Sensitivity to selected antibacterial medicines was investigated using disck method and Mueller-Hinton 2 Agar medium. From the 577 examinated food samples 126 strains of Listeria were isolated and among them: 34.1% L. monocytogenes, 36,5% L. welshimeri, 19.0% L. innocua, 3.17% L. grayi, 0.79% L. seeligeri, 0.79% L. seeligeri/welshinmeri and 5.56% L. ivelshimeri/innocua. L. monocytogenes strains most often were found in minced pork, culinary products and in frozen vegetables. On the base of ApiListeria (bioMeieux) test the isolated L. monocytogenes strains were qualified into 2 biochemical types. It was found that all L. monocytogenes were sensitive to sulphametaksazol/trimetoprim and ampicyllin, 25% of strains were moderatety sensitive to penicillin and only 2 L. monocytogenes strains were resistant to gentamicin. Presence of Listeria spp. microorganisms in food products may be an production hygiene indicator for critical control point and show the possibility of contamination with L. monocytogenes strains. PMID- 17133906 TI - [Influence of selected Lactobacillus sp. on Clostridium difficile strains with different toxigenicity profile]. AB - This study was performed for determination of antagonistic activity of Lactobacillus spp. (L. plantarum 2017405, L. rhamnosus GG, L. acidophilus DSM 21007 and L. fernmentumn 353) on Clostridiunl difficile strains belonging to different toxigenicity profiles. Forty strains C. difficile isolated from patients suffering from antibiotic associated diarrhea (AAD) were used. Among C. difficile strains 13 produced toxin A and B (A+B+CDT-), 14 produced only toxin B (AB'CDT), 9 produced toxins A and B and possessing of binary toxin genes (A+B+CDT ) and 4 were non-toxigenic (A-B-CDT-). We did not observe relationship between degree of antagonistic activity Lactobacillus spp. and profile of toxigenicity of C. difficile strains. PMID- 17133907 TI - [Hydrolytic and haemolytic activity of Klebsiella pneumoniae and Klebsiella oxytoca]. AB - Hydrolytic enzymes and haemolysins are important extracellular substances produced by many bacteria. We investigated 57 K. pneumoniae strains and 40 K. oxytoca strains isolated from clinical materials. We estimated the ability to produce: proteases hydrolyzing milk powder, caseinase, gelatinase, elastase, lecithinase, lipases, DNase and haemolysins on human, sheep and horse erythrocytes on TSA medium with or without 5% Egg Yolk. We detected that K. oxytoca strains produced proteases hydrolyzing milk powder (37.5%), caseinase (15.0%) and gelatinase (17.5%) more frequently than K. pneumoniae strains (respectively 21.0%, 5.3%, 8.9%). None of the analysed Klebsiella spp. strains produced elastase. Only K. pneumoniae strains produced lecithinase (5.3%). Lipases hydrolyzing Tween were produced from 3.5% (for Tween 60 and 80) to 7.0% (for Tween 20). Among K. oxytoca strains only one (2.5%) hydrolyzing Tween 20. DNase was produced by 38.6% of K. pneumoniae strains and by 27.5% K. oxytoca strains. Haemolytic properties on human erythrocytes were detected in 5.3% K. pneumoniae strains on TSA medium and 29,8% on medium with Egg Yolk. In K. oxytoca strains haemolytic properties on human erythrocytes were detected only on medium with Egg Yolk (12.5%). Haemolytic properties on sheep erythrocytes were detected respectively in 21.0% and 22.8% K. pneumoniae strains and in 7.5% K. oxytoca strains on each medium. Haemolytic properties on horse erythrocytes were detected respectively in 33.4% and 52.6% K. pneumoniae strains and in 15.0% and 20.0% K. oxytoca strains. PMID- 17133908 TI - [Genetic diversity analysis of isolates belonging to Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica species]. AB - In this study, Amplified Fragment Length Polymorphism (AFLP) method was used to track differences among human and animal isolates of B. pertussis, B. parapertussis and B. bronchiseptica species. One hundred and sixty representative strains of these species orginated from international and Polish bacterial collections were genotyped according to AFLP involving EcoRI/Msel and SpeI/ApaI restriction/ligation/amplification procedures. This study has confirmed high potential AFLP SpeI/ApaI procedure for intra-species differentiation of B. pertussis and B. bronchiseptica strains. Both AFLP EcoRI/MseI and SpeI/ApaI procedures have been found to be useful for species-specific classification in case of B. pertussis strains. In case of B. bronchiseptica or B. parapertussis species-specific classification, SpeI/ApaI procedure has been found more precise than EcoRI/MseI one. PMID- 17133909 TI - [Susceptibility of fungi isolated from clinical materials to voriconazole]. AB - The objective of the study was evaluation of the susceptibility of 139 fungal strains isolated from clinical materials to Voriconazole, a new antifungal agent of the triazole group. A dilution method was used. The drug was incorporated into the culture medium at concentrations 0.1-100 mg/l. It was found out that the antifungal effectiveness of the drug varied both between various fungal genera and species, and between strains within the same species. Total inhibition of the growth of 20% of the yeast-like fungi and 23.3% of the moulds was achieved at concentration 0.1 mg/l. It was documented that the species Candida guilliermondii, C. kefyr, Saccharomyces cerevisiae and Aspergillus fumigatus, that as a rule are resistant to triazoles, were highly susceptible to Voriconazole. Dermatophytes, too, were highly susceptible to the drug, particularly Trichophyton mentagrophytes and T. rubrum. The drug at concentration 0.1 mg/l totally inhibited 73.5% of the dermatophytes, and at concentration 1 mg/l--97.0% of them after 7 day incubation. After 14 day incubation, 97.0% of the strains were also inhibited at drug concentration not exceeding 1 mg/l. PMID- 17133910 TI - [The influence of AMPs from HaCaT keratinocytes on some bacteria strains derived from skin changes]. AB - The aim of this study was to evaluate antimicrobial activity of protein extracts from HaCaT cell line against bacterial strains, isolated from clinical materials, obtained from patients with clinical symptoms of acne (Propionibacterium acnes) and gas gangrene (Clostridium perfringens and Sterptococcus pyogenes). Reference strain of Staphylococcus aureus ATCC 25923 also was used. Protein extracts from cultured HaCaT cells were obtained by 3-fold freezing/defreezing cells in dry ice following by centrifugation and incubated with appropriate bacterial suspension (0.5 McFarland scale) during 6 and 24 hours. We observed time-depending and strain-depending activity of HaCaT--protein extract. Interestingly, high activity was demonstrated against strains of S. pyogenes and C. perfringens. Because of increasing bacterial resistance to antibiotics further studies in the field of antimicrobial peptides are required. PMID- 17133911 TI - [Possibility to use the lyophilization for the prolongation of the stability of biological compounds used in cosmetic production]. AB - The investigations were conducted with using aquatic hydrolysate of protein 5%, 10%, 20% and 50%. Starting product to prepare solutions with different concentrations, exposed on lyophilisation was pork gelatin (Polonaise, Belgium). The aim of investigations was to determine on effect of lyophilization process on the microbiological and physico-chemistral stability of protein hydrolisate. Physical parameters of lyophilization of protein hydrolysate solutions of different concentrations and physical properties of the lyophilized hydrolysates were determined. Microbiological analysis demonstrated that lyophilization protected the hydrolysates stored at 4 degrees C and 22 degrees C for 60 or 100 days against bacterial contamination. On the other hand, the inhibition of the bacterial multiplication in nonlyophilized hydrolysates was obtained only during storage at -20 degrees C. A proper structure and good dissolubility of the lyophilizated of preparation were obtained for 5%, 10% and 20% hydrolisate and for of the maximum kinematical viscosity 2923 mm2/s. PMID- 17133912 TI - [Costs of occupational diseases and accidents at work in Poland]. AB - BACKGROUND: The costs of occupational diseases and accidents at work in the state members of the European Union keep at a level of several percent of their gross national product (GNP). Employees, employers and the society as a whole have to incur this financial burden. Therefore, all social partners should be involved in the improvement of health and safety in the work environment through their concerted efforts. It should be pointed out that information in the field of economy is an inherent instrument of all activities. It allows to estimate economic consequences of occupational diseases and accidents at work as well as to formulate future strategies. The aim of this study was to estimate and assess the range of the expenditure of money on occupational diseases and accidents at work in Poland in 2004. MATERIAL AND METHODS: The algorithm for estimating economic consequences of occupational diseases and accidents at work was developed, taking into account, e.g., the selected components of the costs, the estimation of economic consequences for the national economy, and the costs incurred by employers and social insurance institutions. In addition, the model for estimating economic consequences and defining the range of expenditure of money on occupational diseases and accidents at work relative to indicators of socioeconomic situation of the country was constructed. Economic consequences are understood as costs incurred by the health care and social insurance systems in Poland, institutions, companies, and individual employees. RESULTS: The following cost components were estimated: the average-annual costs of lost production, sickness benefits, social insurance benefits and those incurred by enterprises related with accidents at work and occupational diseases. CONCLUSIONS: Due to the lack of complete, reliable information essential for estimating economic consequences, a number of assumptions and study limitations were accepted. The range of identified and estimated costs of occupational diseases and accidents at work was set relative to GNP and to the value added in the national economy. Bearing in mind that the costs of occupational diseases and accidents at work are underestimated, it can be concluded that their share in values characteristic of the national economy do not depart from average values recorded in European countries. PMID- 17133913 TI - [Some job factors associated with departure from working life before retirement age]. AB - BACKGROUND: Ongoing demographic changes observed in Poland indicate progressive aging of the population. The process of systemic transformation initiated in Poland in 1989 has contributed to a significant decline in occupational activity among people especially those at the pre-retirement age. The identification of determinants responsible for the decreased occupational activity in older age groups of the population may provide the basis for taking appropriate steps to extend their period of employment, which will become a must in the near future in view of current socio-demographic phenomena. Therefore, the aim of the this study was to identify factors, which mostly contribute to an earlier departure of older workers from working life. The presented analysis concerns occupational factors characteristic of working conditions. MATERIAL AND METHODS: The study was carried out in a group of current and former workers aged over 45 years and below the retirement age, employed at production workposts and in production services in the years 1996-2000. Active workers of the same age (+/- 3 years) and gender, employed at adequate workposts were matched with selected persons who had left their job because of becoming entitled to disability pension, earlier retirement, benefit or pre-retirement allowance. A questionnaire-based interview was used to collect empiric material, and logistic regression to analyze the risk of earlier departure from working life. RESULTS: The investigation carried out in the study group showed that the following factors mostly influenced departure from work before reaching the retirement age: piecework system (OR = 7.76; 95% CI: 2.53 23.79); heavy lifting at work (OR = 1.90; 95% CI: 1.11-3.25); physical workload (OR = 1.92; 95% CI: 0.74-5.00); exposure to low temperature (OR = 1.71; 95% CI: 0.94-3.09); exposure to whole-body vibration (OR = 1.47; 95% CI: 0.96-2.25); subjective assessment of fatigue after work (OR = 2.25; 95% CI: 1.45-3.48); and no leisure time after work (OR = 2.71; 95% CI: 1.52-4.83). CONCLUSIONS: The results of the study can be regarded as an important indication that appropriate preventive measures should be taken to improve working conditions especially for older age groups, or to provide the possibility of their retraining if the occupational activity of older workers is to be increased. PMID- 17133914 TI - [The CIS20R Questionnaire and its suitability for prolonged fatigue studies]. AB - BACKGROUND: The aim of the study was to validate the Checklist Individual Strength (CIS20R) questionnaire for the assessment of prolonged fatigue in workers and to define its psychometric properties and fatigue standards for the population of Poland. The questionnaire contains 20 statements, scored on a seven point scale (1-7), reflecting four aspects of fatigue: subjective feeling of fatigue, reduction of concentration, reduction of motivation, and reduction of physical activity. The outcome yielded by the questionnaire is expressed by the sum of all points scored in responding to all statements and mean values of its components. MATERIAL AND METHODS: The study was carried out in a group (325 men and 363 women) of workers employed in industrial plants, hypermarkets and municipal services; workers of research institutes (81 men and 205 women) and pregnant women (20 persons). Based on the calculated coefficients of correlation between responses to each statement and the total score, strong discriminative power of individual statements was observed. Crombach's alpha was used to assess the consistence of the questionnaire. The coefficient value for the total score was 0.912 and for the fatigue components it ranged from 0.611 to 0.879, which indicates satisfactory reliability of the method. In addition, norms were elaborated using a sten scale. The total value of 40 indicates the borderline between low and moderate and the value of 84 between medium and high level of fatigue. RESULTS: The level of the risk for chronic fatigue was estimated on the basis of investigations among industrial, trade and municipal services workers. In the group of men, the necessity of prolonged standing during work, high physical effort during work, lack of fitting to physical effort during work, and stress connected with responsibility (component of occupational stress) were the risk-increasing factors. In the group of women, they were: lack of fitting to physical effort during work, noise disturbing reasoning of speech and stress connected with psychic load and lack of social support (components of occupational stress). In both groups, sleep disturbances proved to be responsible for increasing risk of the incidence of chronic fatigue. In the group of men--the diet, and in the group of women - exercise during leisure time were found to be protective factors. CONCLUSIONS: The study revealed that the CIS20R questionnaire is a good research tool to study chronic fatigue. It was also found that the level of fatigue in the population of Poland is much higher than that in the Netherlands, the country where the questionnaire has been developed. In the Dutch studies, the value of 76 was found to be a borderline between "normal fatigue" and fatigue occurring in people with impaired health. In our studies this value corresponds with six stens, which means that it is still within the range of moderate values. PMID- 17133915 TI - [The most common faulty postures among boys aged 13-16 years measured by Moire's photogrammetric method]. AB - BACKGROUND: The project concerns faulty postures. The aim of the study was to evaluate postures among boys aged 13-16 years by Moir's photogrammetric method. MATERIAL AND METHODS: The study covered 191 boys, including 52 pupils aged 13 years attending the Primary School No. 11 in Starachowice, and 139 students (49, 45 and 45 boys aged 14, 15, and 16 years, respectively) of the Grammar School No.3 in Starachowice The study was carried out in April 2004. RESULTS: The most common lateral curvanture of the spine was observed in 131 (69%) boys, posture asymmetry in 60 (15%) boys, among defects in the sagittal plane concave back was found in 28 (15%) boys, flat back in 24 (13%), and round back in 2 boys aged 16 years. CONCLUSION: In view of a large proportion of lateral curvanture of the spine (69%), there is an urgent need to intensify prophylaxis and medical treatment in teenagers. Moreover, the issue of body posture needs further investigations. PMID- 17133916 TI - [HIV infection under laboratory conditions]. AB - Laboratory workers are at a particularly high risk of acquiring HIV. Based on the medical literature, selected cases of occupational HIV infections among laboratory workers are presented. Having analyzed specific circumstances connected with occupational exposures, risk factors of such incidents are discussed. The importance of continuing education in the areas of infection control procedures and compliance with universal precautions as well as reporting on occupational exposures to any infectious material in the context of post exposure prophylaxis are pointed as the best ways to achieve a successful outcome in the HIV infection prevention under laboratory conditions. The lack of efficient, multifaceted legislation covering all aspects of occupational exposure to blood-borne pathogens, still observed in Poland, is stressed. PMID- 17133917 TI - [Occupational allergy caused by ornamental plants]. AB - The problem of allergy to decorative plants is still poorly known. Reports on occupational allergy to flowers are scarce and usually concern gardeners, greenhouse workers and florists. The handling, smelling and caring of flowers may cause rhinoconjunctivitis, asthma, urticaria and also contact dermatitis. Plants of the Compositae family that includes many different species have been frequently described. Chrysanthemum flowers, the strongest sensitizer among ornamental Compositae plants, especially contribute to the development of contact dermatitis. Allergy to Liliaceae, mostly to tulips, hyacinths, lilies and crocuses has quite often been reported as well as sensitization to rose pollen in rose planters. Occupational sensitization to flowers of other families, among which spathe flowers, primulas, weeping fig or Stephanotis floribunda should be mentioned, is less frequent. Exposure to ornamental flowers is common in the general population. Persons occupationally involved in cultivation of flowers and who demonstrate allergic symptoms are often forced to change their jobs. Candidates to these occupations with diagnosed atopy should be informed about the risk of developing allergy to flowers, which could make them unable to perform the job. PMID- 17133918 TI - [Influence of environmental risk factors on the development of chronic vein insufficiency]. AB - In European countries, vein diseases pose a serious social problem; as many as 35% of professionally active population, 50% of pensioners, and every second female suffer from this disease. Chronic vein insufficiency (CVI) is one of the most frequent forms of vein diseases, particularly in developed countries; it is thought to concern one third of the population. This paper presents basic information on CVI epidemiology, etiopathogenesis and pathophysiology and also reviews the literature on the following CVI risk factors: obesity, lack of physical activity, dietary habits, including constipation, pregnancy, genetic factors as well as prolonged sedentary or upright position during job performance. PMID- 17133919 TI - [Risk bloodborne infections in health care workers]. AB - The paper presents current data on epidemiology and risk factors responsible for incidents leading to blood-borne infections among health care workers. In many countries, the number of this type of incidents has markedly decreased, whereas in Poland blood-borne infections are still a serious problem. Unfortunately, the circumstances, in which such incidences happen, are frequently caused by oversight and typical mistakes. The problem also lies in that not all cases are regularly reported. Therefore, it is essential to draw special attention to under and post-graduate programs covering all medical professions, in which the problem of occupational and hospital infections should be an obligatory element of training. PMID- 17133920 TI - [Petroleum substances--human health hazards. I. Classification of petroleum substances on the list of dangerous substances and assessment of existing hazards]. AB - Complex products derived from petroleum are widely used as fuels, greases, solvents, and intermediates in many branches of industry. Petroleum exposure related human health hazards, observed in occupationally exposed people and in the general population, are a serious sanitary problem. Complex and variable composition of individual petroleum products makes the actual assessment of human health hazards difficult. Potential hazards, and resulting classification of individual petroleum substance groups, are discussed in the presented work. This should prove to be helpful to work safety and hygiene services as well as to supervising institutions, mainly the sanitary inspection, in a proper assessment of the hazards, and consequently in taking appropriate preventive actions. In Part I., general issues concerning the hazard assessment and legal aspects of petroleum substances classification are presented. In Part. II., individual groups of petroleum substances are discussed with respect to health hazards, resulting from both physicochemical properties and toxicity, and their classification based on this analysis is suggested. PMID- 17133921 TI - [Secondary nephropathies in occupational health practice. I. Secondary nephropathies due to occupational exposure]. AB - Kidneys are particularly susceptible to the effect of toxic agents that can cause renal damage and even renal failure. The aim of this study was to present current knowledge of the influence of occupational toxic agents on renal functions. The authors describe the major nephrotoxic factors, such as heavy metals (including lead, cadmium, and mercury), organic solvents, pesticides and silica. They discuss changes in the kidney structure and molecular mechanisms of nephrotoxicity and carcinogenesis induced by occupational exposure to major toxic agents and also indicate possibilities of detecting the predicted nephrotoxicity symptoms so that clinical, overt renal insufficiency could be prevented. PMID- 17133922 TI - Is your healthcare environment therapeutic? PMID- 17133923 TI - Best practice in the assessment and management of diabetic foot ulcers. AB - Diabetes is an increasingly serious health issue in the rehabilitation population. Foot ulcers develop in approximately 15% of people with diabetes and are a preceding factor in approximately 85% of lower limb amputations. Nurses have significant opportunity to positively influence client outcomes and quality of life by promoting maintenance of healthy feet, identifying emerging problems, and supporting evidence-based self-care and interdisciplinary intervention. Best practice guidelines (BPG), such as those developed by the Registered Nurses Association of Ontario, provide a framework to enhance nursing practice and promote excellence in client care. This article highlights key evidence from the BPG, "Assessment and Management of Foot Ulcers for People with Diabetes," and other relevant diabetes literature. This information better equips rehabilitation nurses to promote ulcer prevention strategies; identifies key factors in ulcer risk; and utilizes current, best evidence for ulcer assessment, management, and evaluation. PMID- 17133924 TI - Physical restraint reduction in the acute rehabilitation setting: a quality improvement study. AB - A prospective, continuous quality improvement study was implemented at a hospital on two rehabilitation units: stroke and brain injury. The purpose of the study was to decrease restraint use by 25% and to maintain fall rates no greater than 10% over baseline. A multi-component restraint reduction program was implemented that focused on administrative support, education, consultation, and feedback. Monthly restraint rates and fall rates were monitored and compared to the previous year's rates. Both units reduced restraint use. Importantly, this reduction was accomplished at the same time as a decline in fall rates. PMID- 17133925 TI - Evaluation of the research agenda for rehabilitation nursing. AB - From 1993 to 1995, under the direction of the board of directors of the Association of Rehabilitation Nurses (ARN), the Rehabilitation Nursing Foundation (RNF) created a research agenda for the field, which was published in 1996. The goal of the agenda was to identify a broad range of issues in rehabilitation practice that could be examined to advance the scientific knowledge base. Ten years later, the board of directors of ARN directed RNF to evaluate and update the agenda. A committee of three ARN members was formed to undertake this task. This article reports the committee's evaluation of the 1996 Rehabilitation Nursing Research Agenda. A separate article reports on the process of revising the agenda. PMID- 17133926 TI - Spiritual care: practical guidelines for rehabilitation nurses. AB - Spiritual care has long been recognized as an essential component in providing holistic care to patients. However, many nurses have acknowledged that their education lacked practical guidelines on how to provide culturally competent spiritual care. Although all nurses are required to provide spiritual care, rehabilitation nurses are particularly challenged to be competent in this area, due to the lengthy recovery time and special needs often presented by rehabilitation patients. This article provides practical guidelines for rehabilitation nurses, to assist patients in meeting their spiritual needs. PMID- 17133927 TI - Nursing interventions within the Mauk Model of Poststroke Recovery. AB - Stroke continues to be the third leading cause of death in the United States. According to the National Stroke Association (NSA, 2004) and the American Heart Association (AHA, 2004), there are over 750,000 new or recurrent strokes per year, with many resulting in residual disability. Stroke survivors often deal with the physical, psychosocial, and emotional consequences of stroke long after they have left the safety of professional rehabilitation. Patient instruction from nurses prior to discharge, while necessary, may be done at a point in the recovery process when the stroke survivor is not ready to learn how to deal with such consequences. Using the Mauk Model for Poststroke Recovery, nurses can identify which phase of recovery a survivor is in, and thus tailor care to his or her needs. The purpose of this article is to use the Mauk Model for Poststroke Recovery to present nursing interventions that are appropriate to each of the previously identified six phases of stroke recovery. PMID- 17133928 TI - Temporary restorations in endodontics: a review. PMID- 17133929 TI - Reaching your implant potential. PMID- 17133930 TI - Diagnosis and treatment of molar incisor hypomineralization. AB - Molar incisor hypomineralization (MIH) is a relatively common condition that varies in clinical severity and can result in early loss of the permanent 6-year molars. The etiology of MIH remains unclear, and the diagnosis can be confused with more generalized enamel defects such as those that occur in amelogenesis imperfecta. The management of MIH depends largely on the severity of the enamel defect. Degrees of hypomineralization can range from mild enamel opacities to enamel that readily abrades from the tooth as it emerges into the oral cavity. Usually, severely affected molars are extremely hypersensitive, prone to rapid caries development, and can be difficult to manage in young patients. The purpose of this article is to review approaches to diagnosing and treating MIH. PMID- 17133931 TI - Patient and parental perception of a new fluoride varnish. AB - The purpose of this study was to compare children's and their parents' acceptability of a new fluoride varnish that purports to have improved taste and color with a control varnish. Sixty patients, 4 to 17 years of age, attending recall appointments at the University of Washington Pediatric Dental Clinic, Seattle, Washington, were randomly assigned to receive 1 of 2 fluoride varnishes. After their appointments, patients and their parents were asked about the varnish's taste and color. There were 32 patients 4 to 7 years of age and 28 patients 8 to 17 years of age. All 8- to 17-year-olds agreed that "white varnish" was preferable to "brown varnish." Most of the 4- to 7-year-olds also agreed that white was better. The 4- to 7-year-olds did not like the taste of either varnish, but the 8- to 17-year-olds preferred the taste of white. All parents cared mostly about efficacy, but agreed that the white, better-tasting varnish was a positive improvement. Parents and their children agreed that though the white varnish is best, the taste was far from ideal for the younger children, based on its single bubble-gum flavor and the standard control fluoride varnish available. PMID- 17133932 TI - Rapid onset of Parkinsonian-like symptoms in a geriatric dental patient: a case report. AB - This article reviews the clinical issues regarding adverse drug reactions in geriatric dental patients. Accurate clinical observations and diagnosis are complicated in geriatric patients because they are predisposed to chronic illnesses, various prescribing physicians, and a decreased ability to metabolize and detoxify multiple medications. The authors have further reviewed neurological motor reactions with a detailed review of the physical presentations of Parkinson's disease. As such, the dental professional has a unique opportunity to provide observational feedback to other healthcare providers concerning the health status of their geriatric patients. In this case report, the changes in the patient's physical status and mental well-being were not a result or associated with a catastrophic event (eg, stroke, cardiovascular event, or head injury). The patient's rapid degeneration was anecdotally associated with a recently prescribed group of medications, which shows the need for healthcare professionals to be aware of changes in medications when assessing patients' health. PMID- 17133933 TI - Parenting the parents of pediatric patients. AB - Managing the behavior of pediatric patients has been the focus of many studies; however, paying attention to the parents and their individual circumstances may be just as important to achieve our treatment goals. Parents today expect to be involved in treatment decisions and often have distinct expectations regarding their children's experiences. Parental attitudes and emotions can adversely affect the tone of the appointment and negatively influence the child's perception of the event. Managing the behavior of a challenging child is difficult, but combining that with the challenging behavior of a parent can be exhausting. This article will provide anticipatory guidance on how to enhance communication skills to avoid confusion and offer direction to parents before, during, and after the treatment of their children. PMID- 17133934 TI - [Future prospects]. AB - The immediate prospects of development of radiodiagnostic equipment on the basis of digital systems for medical image representation and processing are considered. Special attention is paid to development of X-ray diagnostic systems. PMID- 17133935 TI - [Two technologies for designing digital X-ray image detectors]. AB - The goal of this work was to discuss the problem of replacement of conventional devices for film roentgenography with modern digital X-ray image detectors. Several types of digital detectors are considered. The material presented in this work makes it possible to draw some conclusions on the prospects of development of X-ray detectors and transducers, their advantages and disadvantages. PMID- 17133936 TI - [Digital roentgenographic systems based on photostimulated screens and their comparison with roentgenographic screen-film sets]. AB - Parameters of images obtained using roentgenographic systems based on stimulated luminophores (computer roentgenography) were experimentally compared. It was shown that roentgenography based on the use of stimulated luminophores had several advantages over conventional film roentgenography. PMID- 17133937 TI - [Experimental and computational evaluation of quality of digital X-ray imaging]. AB - The results of the use of specialized software for experimental and computational evaluation of the characteristics of digital X-ray detectors are analyzed. PMID- 17133938 TI - [Comparative analysis of dose distribution in photon and proton therapy of prostate cancer]. AB - Comparative analysis of the dose distribution in photon and proton therapy of prostate cancer is performed. The degree of exposure of healthy organs to radiation is assessed for each of the two treatment techniques using the time dose-fractionation parameter. PMID- 17133939 TI - [A heterogeneous air-filled ionization chamber without external power supply]. AB - Possibility of the use of heterogeneous ionization chambers with air-filled interelectrode space for X-ray detection is discussed. Qualitative analysis of the process of generation of ionization current in the heterogeneous chamber without external power supply is performed. Experimental data presented in this work show that ionization chambers with triple heterogeneous structure provide high operating characteristics when connected in active integrator and current-to voltage converter circuits (especially in the case of differential connection circuit). PMID- 17133940 TI - [Analysis of frequency-contrast characteristics of digital X-ray image detectors]. AB - The effect of various computing procedures on the results of estimation of parameters used for MTF calculation is analyzed. Experimentally evaluated modulation transfer functions of some types of digital X-ray image detectors are presented. PMID- 17133941 TI - [A method for calibration of spectrozonal X-ray diagnostic system]. AB - A method for calibration of X-ray diagnostic system equipped with a spectrozonal X-ray detector is described. Analytic expressions for calculation of the effective atomic number of the substance of inclusion tissue revealed in spectrozonal X-ray images are obtained. PMID- 17133942 TI - [Specific features of design of digital X-ray diagnostic apparatuses on the basis of CCD matrices]. AB - Specific features of modern digital screen-optics-CCD-matrix systems are considered. Characteristics of single- and multi-matrix systems are analyzed. It is shown that all disadvantages of single-matrix systems as compared to multi matrix systems can be eliminated using modem high-efficiency CCD matrices. PMID- 17133943 TI - [An in vitro study of low-frequency ultrasound transmission through a musculoskeletal fragment of thoracic cage]. AB - Low-frequency ultrasound transmission through a musculoskeletal fragment of thoracic cage was studied in vitro. The degree of ultrasound attenuation during noninvasive transthoracic application was estimated. The mean coefficient of ultrasound attenuation in thoracic cage was determined for the low-frequency region (210 kHz). The temperature effects of ultrasound application were also studied. PMID- 17133944 TI - [A new method for examination of human skin vasomotions]. AB - An original method for determination of elasticity of biological tissues from their resonance frequency has been devised. The method can be used for detection and spectral analysis of small oscillations of skin elasticity caused by vasomotions. PMID- 17133945 TI - [Synthesis of radiological models and radiological constants. Part 4: Synthesis of population-phenomenological models and Lyman model]. AB - The goal of this work was to synthesize Lyman mathematical models (MM), which describe the probability of post-radiation complications (PRC) in tissue subjected to radiation therapy with given scheme of dose fractionating (DF), and population-phenomenological (PP) MMs PP3 and PP4, which describe equivalent DF schemes for a fixed PRC value. Construction of synthesized MMs (SMMs) becomes possible only on the basis of several assumptions requiring further clinical validation. Synthesized MMs can be used for determination of the optimal dynamic conditions of irradiation of malignant tumors. These conditions include the optimal physical plan of irradiation and the optimal time scheme of its implementation. Synthesis of MMs leads to determination of radiological invariants (constants), which can become a basis for a new branch of medical science, quantitative radiology. PMID- 17133946 TI - [Problems of organization of antituberculosis prophylaxis in rural areas of the Russian Federation]. AB - Possibility of use of mobile fluorographic rooms based on KAMAZ trucks for mass scale prophylactic examinations of rural population of the Russian Federation is discussed. The mobile rooms are equipped with digital fluorographic systems. A list of the main medical and performance characteristics of these systems is presented. An example of equipment location in a mobile room is given. PMID- 17133947 TI - [A study of digital x-ray detection systems with optical image transfer]. AB - The work describes the performance characteristics of roentgenographic screen optics-CCD-matrix systems based on the KRTs-1 digital roentgenographic system developed at SpektrAp, Ltd. The efficiency of X-ray detection is estimated from the minimum detectable contrast and maximum resolution. The effect of the X-ray imaging system components on the efficiency of detection is studied. The X-ray screen structure is shown to have a crucial effect on the efficiency of detection as estimated from the image resolution. PMID- 17133948 TI - [Experience in the use of electron KRT remote-controlled X-ray diagnostic system]. AB - Short description of the Electron KRT system and its performance is given. Functional capabilities of the system are considered on the basis of the accumulated operating experience. PMID- 17133949 TI - Development and plasticity of the retina in the opossum Monodelphis domestica. AB - We investigated the rate of cell proliferation and death in the retina of the Monodelphis opossum during its postnatal development and the influence of early monocular enucleation on these processes. Our results show that in the opossum, as in other marsupials, the peak of the retinal cells divisions occurs postnatally and that generation of retinal cells continues till the time of eye opening (P34), except of the marginal rim, where it continued till P60. Ganglion and amacrine cells are generated between postnatal days (P) P4 and P9, while bipolar cells and photoreceptors are generated simultaneously between P14 and P25. The peak of ganglion cell death as detected by the TUNEL method occurs around P14-19 in the center of retina. The second peak of apoptosis appears in the inner nuclear layer (INL) at P19-25. Gliogenesis takes place between P25 and P34. We also found that monocular enucleation performed during the early period of retinal development (P0-P7) did not influence proliferation, developmental apoptosis or other developmental processes in the retina of the remaining eye. PMID- 17133950 TI - Are there different mechanisms of synchronization in the course of spike-wave discharges (SWDs) burst development in WAG/Rij rats? AB - In WAG/Rij rats the pair linear correlation r was calculated for bipolar recordings in fronto-temporal, fronto-occipital and occipito-temporal zones of both hemispheres as well as in paleocerebellar cortex (culmen). It was shown that development of SWD bursts resulted in interhemispheric decreases of correlation between the right occipito-temporal cortical region on one side, and left fronto temporal on the contralateral side. Towards the end of SWD, we found an increased interhemispheric correlation between left fronto-temporal and right fronto occipital cortical zones, as well as, between both fronto-temporal zones. Paleocerebellum correlates at a weak to moderate level during different periods of SWD burst generation. PMID- 17133951 TI - Propagation of EEG activity during finger movement and its imagination. AB - We investigated the pattern of EEG activity propagation in the beta and gamma band during a finger movement experiment and imagination of that task. The data were analyzed by means of a short-time directed transfer function (SDTF) based on a multivariate autoregressive model. The signals from the right (or left) hemisphere were processed simultaneously (not pairwise), which is crucial for obtaining a correct picture of EEG activity transmissions. The pattern of propagation in the beta band involved for both tasks a decrease of the propagation from the motor areas during the execution of the movement - less pronounced in the case of imagination. The performance of the motion was mainly connected with a short outburst of gamma activity from the hand sensorimotor areas. In case of imagination the gamma outflow lasted longer and concerned larger brain areas. PMID- 17133952 TI - Photoperiod affects distribution of dynorphin A in the brain of Siberian hamster. AB - Dynorphin A1-77 (DYN A1-17) acting in the CNS is known to affect thermoregulation, water and energy balance in the short time scale. In this study a long-term alteration of these functions induced by changes of day length in the highly photoperiodic species, the Siberian hamster (Phodopus sungorus) was studied using immunohistochemistry for DYN A1-17. We found that in the long day (LD, L:D 16 h:8 h) more brain areas express DYN A1-17 peptide than in the short day (SD, L:D 8 h:16 h) conditions. Structures of the hypothalamo-pituitary axis as well as cells of the ependyma, subcomissural organ and choroid plexus of the lateral and third brain ventricles are immunoreactive to anti-dynorphin IgG only in the LD. This might indicate a seasonal regulatory role of DYN A1-17 in physiological adaptations to severe climate changes. PMID- 17133953 TI - Influence of dizocilpine (MK-801) on neurotoxic effect of dexamethasone: behavioral and histological studies. AB - Elevated levels of endogenous glucocorticoids (GCs) or prolonged treatment with high doses of dexamethasone (DEX) or other GCs preparations are frequently associated with psychosis as well as cognitive deficits, such as the impairment of memory and learning. GCs potentiate stress or ischemia-induced accumulation of excitatory amino acids in the extracellular space of hippocampus. The antagonism of glutamate receptors may potentially improve safety profile of therapy with GCs. The purpose of the present study was to investigate the effect of dizocilpine maleate (MK-801), non-competitive NMDA glutamate receptor antagonist, on neurotoxic effect of the prolonged treatment with the high dose of DEX. The results showed that DEX (120 mg/kg/day for 7 days) impaired the long-term memory and the motor coordination, reduced the body weight and induced the lethality of mice. The morphological and ultrastructural study have confirmed damage to hippocampal neurons especially in the CA3 region after the prolonged treatment with DEX alone. Damaged pyramidal neurons showed robust changes in the shape of the nucleus and cytoplasm condensation. MK-801 alone (at non-toxic dose of 0.3 mg/kg/day), changed neither the behavior of mice nor morphology of the hippocampal neurons. However, it did not prevent the neurotoxic effects of DEX. On the contrary, it intensified DEX-induced neurotoxicity. PMID- 17133954 TI - The acute effects of 3-nitropropionic acid on the behavior and spontaneous cortical electrical activity of rats. AB - In this study, the acute effect of 3-nitropropionic acid was investigated on open field and startle behavior of rats, and on their cortical electrical activity. Spontaneous locomotor activity, acoustic startle response, and pre-pulse inhibition of acoustic startle were measured in male Wistar rats (10 weeks old, 180-200 g body weight) after a single dose of 10 or 20 mg/kg i.p. 3 nitropropionic acid. After the behavioral tests, the rats were anaesthetized, and spontaneous cortical electrical activity was recorded. The vertical, horizontal and local open field performance showed dose-dependent deterioration in the rats treated with 3-nitropropionic acid. The number of "noise-positive" startle responses showed non-significant changes, but the inhibition by pre-pulse was significantly reduced in the high dose animals. High dose also increased the proportion of low-frequencies in the cortical activity. 3-nitropropionic acid, known primarily to act in repeated doses (e.g., in animal models of Huntington's disease) had also some clear-cut acute effects on behavioral and electrophysiological parameters of the treated rats. PMID- 17133955 TI - Response to novelty of various types in laboratory rats. AB - One purpose of the two studies reported here was to examine the reactions of rats to changes in the spatial arrangement of a familiarized environment under low stress conditions. The second purpose was to test the role of rats' experience with novelty. In Experiment I the novelty was manipulated by introducing new tunnels into one zone of the experimental chamber. The introduction of novelty took place after 11 habituation sessions. In Experiment II in the course of habituation sessions the experimental group of rats was subjected to a persistent change of tunnel arrangement in the experimental zone, whereas for the control group nothing changed. All animals reacted to the novelty with increased time spent in the experimental zone and decreased time spent in other zones. Both experiments show that under low stress conditions rats demonstrate a positive response toward novelty and that their previous experience with novelty does not affect that reaction. PMID- 17133956 TI - Asymmetry of the discrimination function for temporal durations in human subjects. AB - Ten human subjects were comparing durations of pairs of visual stimuli in a two way forced-choice task. Mean durations of presented time intervals were -3 s ("short") or -6 s ("long"); the duration ratio was varied at nine levels. The Weber fractions for the short and long durations were approximately equal, -0.22. The ratio of subjective equality was almost exactly unity for the short durations, but it was significantly reduced (-0.76) for the long durations. This asymmetry of the discrimination function indicates time-dependent change of internal representations of past durations, and is well compatible with the "dual klepsydra model". Model-based estimates of the internal time representation loss rate, derived from the present data, are in a good agreement with values obtained from earlier studies on duration reproduction. PMID- 17133957 TI - Non-vagal apnea evoked by intra-common carotid artery injection of N-methyl-D aspartic acid (NMDA) in anesthetized rats. AB - Respiratory effects of an intra-common carotid artery injection of N-methyl-D aspartic acid (NMDA) were investigated in anesthetized spontaneously breathing rats, using three experimental paradigms: (1) midcervical vagotomy followed by supranodosal vagotomy, (2) midcervical vagotomy followed by section of the carotid sinus nerves (CSNs), and (3) midcervical vagotomy followed by pharmacological blockade of NMDA receptors. The intra-common carotid artery injection of NMDA (4 mg/kg) induced transient expiratory apnea followed by a brief and variably occurring period of breathing at reduced tidal volume. There were no consistent changes in respiratory rate in rats subjected to midcervical vagotomy alone. Supranodose vagotomy exerted no effect on NMDA-induced respiratory arrest, whereas CSNs' section or blockade of NMDA receptors with AP-7 abolished the apnea. These results indicate that the apnea induced by intra arterial NMDA challenge is due to activation of peripheral NMDA receptors and is mediated via carotid body afferents. PMID- 17133958 TI - Focal stroke in the barrel cortex of rats enhances ipsilateral response to vibrissal input. AB - Brain injury triggers spontaneous plasticity, often resulting in considerable restoration of function. To investigate mechanisms of this compensatory plasticity we followed changes in the brain's pattern of activation evoked by stimulation of vibrissae, after a focal cortical stroke which destroyed the cortical representation of vibrissae, the barrel cortex. The pattern of brain activation was visualized with [14C]-2-doexyglucose (2DG) autoradiography in rats 7 days after photothrombotic stroke. During isotope incorporation, vibrissae contralateral to stroke were stimulated. In control rats this stimulation activates the barrel cortex and the second somatosensory cortex in the contralateral hemisphere. Seven days after stroke in the barrel cortex, significant increases in activation were found in ipsilateral, uninjured hemisphere in the barrel cortex and anterior vibrissae representation, and also in regions not specifically connected to vibrissae stimulation, such as motor and auditory cortex. Shortly after cortical stroke, the intact hemisphere shows higher metabolic activation in several cortical regions, possibly due to abnormal interactions with the injured hemisphere. PMID- 17133959 TI - [Neonatal mortality in a regional perinatal hospital in Merida, Yucatan, 1995 2004. II. Analysis of causes and risk factors]. AB - OBJECTIVE: To analyze the causes of neonatal mortality, its distribution, the moment of appearance and risk factors. PATIENTS AND METHOD: We studied a cohort of 46,297 live newborns born at the mentioned hospital, weighing 500 g or more. In a database we registered: birth weight, gestational age, days of hospital stay, discharged condition, one and five Apgar minute score, morbidity, death causes, moment of death, reducible death/reducible difficult death rate presented in less than 24 hours, 1 to 6 days, and 7 to 27 postnatal days. We compared morbidity, mortality, and risk factors in a three-year period to facilitate the analysis. RESULTS: Mortality increased with the lesser one minute Apgar score: 0.2, 9.7 and 42.9% when score was 7-10, 4-6, or 0-3, respectively, and 0.6, 41.9 and 62.9% at five minute Apgar score. Malformations were the first cause of death, which increased from 28.6 to 40.3%. Respiratory distress syndrome mortality decreased 34% and that of meconium aspiration syndrome 53%. Reducible death/hardly reducible death rate occurred from 1 to 6 days decreased 67%, from 5.2 to 1.7 (reference value 1). CONCLUSIONS: Preventable causes of death decreased significantly, particularly those related to perinatal causes, although they are still high compared with developed countries. PMID- 17133960 TI - [Thyroid dysfunctions and pregnancy]. AB - OBJECTIVE: To study thyroid dysfunctions during pregnancy. PATIENTS AND METHOD: Descriptive, transversal and retrospective study of 61 thyroid dysfunctions during pregnancy, from January 1982 to December 2002. RESULTS: There were 44 hyperthyroidism (1 in 1,908 deliveries: 0.05%) and 17 hypothyroidism (1 in 4,939: 0.02%). Main findings in hyperthyroid patients were: 30-34 years and 2 gestations (34.11%), arterial hypertension (15.90%), abortions (9.09%), preterm delivery (9.09%), term delivery (56.82%), cesarean section (25%), medical treated (97.78%), fetal weighs 3,000-3,499 g (25.61%), 1 and 5 minutes Apgar 8-10 points (60.54 and 81.59%), perinatal mortality rate 0.07 per 1,000 and there was no maternal mortality. And in hypothyroid women: 25-29 years (64.72%), one gestation (29.44%), abortions (17.64%), premature rupture of membranes and oligoamnios (17.64%), term delivery (35.28%), medical treated (100%), fetal weighs 2,500 2,999 g (56.25%), 1 and 5 minutes Apgar 8-10 points (78.58% and 78.58%) and there was no preterm delivery and perinatal or maternal mortality. CONCLUSIONS: Thyroid dysfunctions are infrequent in pregnancy and have little repercussions in perinatal morbidity and mortality. PMID- 17133961 TI - [Physical activity and inactivity among female health service workers]. AB - OBJECTIVE: To determine the prevalence of physical activity and inactivity among female health service workers. PATIENTS AND METHOD: Across-sectional study was conducted during the January-July 2005 period. We included 224 (37%) workers of the General Hospital of the Instituto Mexicano del Seguro Social in Colima, Mexico. We carried out a measurement of the physical activity and inactivity. The indicator was obtained from the international physical activity questionnaire. The obtained data were captured in a computer terminal and processed by means of an automated model. We calculated the frequencies distribution and the prevalence. RESULTS: From the 224 women, 24 (11%) carried out intense physical activity, 22 (10%) moderate, 81 (36%) insufficient, and 97 (43%) qualified as sedentary. CONCLUSIONS: The physical activity and inactivity prevalence among the female health service workers was of 21 and 79%, respectively. PMID- 17133962 TI - [A descriptive study of psychiatric population at Instituto Nacional de Perinatologia]. AB - OBJECTIVE: To establish a description of the psychiatric population of the Instituto Nacional de Perinatologia (INPer), as well as of the exposed newborns to psychopharmacological treatment during the gestation. MATERIAL AND METHOD: One hundred forty-six women were gathered, by means of serial files; 85 of them had an obstetric episode of high risk and 61 were assisted in the institution due to gynecological alterations. A clinical psychologist and a psychiatrist carried out, each one, an interview and the clinical history of all the women. Once the type diagnosis was determined (DSM-IV), they received psicotherapeutical, pharmacological or mixed treatment, until the resolution of the obstetric episode and the gathering of the neonatal data. RESULTS: The findings reported that the prevalence or mental disorders (DSM-IV) that are not adaptive at the INPer is 1.8%. The diagnostic distribution of the psychiatric population ranked in first place the affective disorders (63.7%), particularly major depression disorder (42.5%). These percentages correspond at the institutional total population's 1.19 and 0.79. As for the treatment type, 57 women received psychotherapy, 75 received psychopharmacological treatment and 14 received both treatments. The newborn babies of obstetric patients who received psychopharmacological treatment did not show differences in Apgar, weight, size, complications or obstetric resolution in comparison with patients who received psychotherapy and again with respect to institutional media. DISCUSSION: The prevalence found in the Institution is significantly inferior to that reported at a national and global level, due to the use of two diagnostic interviews by mental health specialists. The wrong use of the sieve is one of the reasons for overdiagnosis in mental health. PMID- 17133963 TI - [Maternal and fetal morbidity in obese pregnant women]. AB - OBJECTIVE: To determine the maternal and fetal morbidity in obese pregnant women compared with non-obese pregnant women. PATIENTS AND METHODS: It was carried out a case-control study. There were included 342 patients who had a body mass index previous to the pregnancy of 18.5 to 24.9 (control group) and 342 pregnant women with body mass index > 30 (group of obese women). We registered the mother and newborns' data to evaluate their morbidity. The groups were compared with Student's t test or Mann Whitney's U test for continuous data and chi-square or Fisher exact test for categorical variables. RESULTS: We found more macrosomic newborns in the group of obese women (p = 0.003) and a higher number of caesarean sections (48.8 vs 37.4%, p = 0.003). The maternal morbidity characterized by gestational diabetes was higher in the obese ones (3.5 vs 0.58%, p = 0.015). Other variables as preterm delivery, stillbirths, malformations, admissions to the neonatal intensive care unit, as well as the development of hypertensive disorders of pregnancy were not significant. CONCLUSION: We found higher maternal and fetal morbidity in obese women. Therefore, these patients should be considered as carriers of high-risk pregnancies. This strategy could avoid complications associated to this group of patients. PMID- 17133964 TI - [Responsible procreation]. PMID- 17133965 TI - [Hypogonadism caused by Gorlin-Goltz syndrome]. AB - The Gorlin-Goltz syndrome is a dominant autosomic disorder characterized by cancerigenic predisposition and multiple development defects, apparently without reproductive compromise. The complex is characterized by four primary symptoms, which include nevoid basal cell epitheliomas malignantly prone, keratocystic jaw, skeletal abnormalities and intracranial calcifications. Apparently, reproductive problems reported had been rarely associated with this syndrome. We present the case of a patient with clinic stigmatae of Gorlin-Goltz syndrome, who had a characteristic progress as seen in the literature; he was the fifth product of a 43 year-old female (father was 48 years old); who at birth disclosed right eye microftalmy, bilateral cryptorchidism surgically treated at age of six. At puberty, an odontogenic cyst of the jaw was noted and enucleated. He also showed facial nevi in neck, thorax and abdomen. When he was admitted being 14 years old in our clinic, he had recurrent bilateral cryptorchidism, sexual immatturity and infertility. It is important to take into consideration Gorlin-Goltz stigmatae in cases of hypogonadism in order to recognize a further genetic influence. PMID- 17133966 TI - [Vulvar cellular angiofibroma. A report of a case and bibliographic review]. AB - The vulvar cellular angiofibroma is a rare mesenchymal tumor. It can be located in many places, but it is more frequent in the vulvar area. It is characterized for being of superficial and slow-growth and for having low propensity of local recurrence. It was described in 1997 by Nucci and it had been reported 51 cases worldwide, from which 18 had vulvar location. This is the first case in Mexico City. It was found in a 36 years-old woman. The specimen was processed by structural and immunohistochemical analysis. Misdiagnosis is common and it can be confounded with spindle cell lipoma, hydrocele of the canal of Nuck, fibromas, angiomyofibroblastoma and many other mesenchymal tumors. It should be distinguished from the aggresive angiomixoma. To improve the clinical diagnosis and treatment, it is very important to have in mind this relatively new entity. PMID- 17133967 TI - [Surgical-abdominal problems during gestation, excluding genital organs. 1951]. PMID- 17133968 TI - A jurisprudence of dysfunction: on the role of "normal species functioning" in disability analysis. PMID- 17133969 TI - "Conscience clauses" or "unconscionable clauses": personal beliefs versus professional responsibilities. PMID- 17133970 TI - Executions and torture: the consequences of overriding professional ethics. PMID- 17133971 TI - A world less silent: celebrating Jay Katz's contributions to law, medicine, and ethics. PMID- 17133972 TI - The healing wisdom of Jay Katz. PMID- 17133973 TI - The uses of psychoanalysis in law: the force of Jay Katz's example. PMID- 17133974 TI - Experimentation with human beings: light or only shadows? PMID- 17133975 TI - The web of relations: thinking about physicians and patients. PMID- 17133976 TI - [Current status of poisoning of companion animals in veterinary science]. PMID- 17133977 TI - [Clinical poisonings and genomics]. PMID- 17133978 TI - [Clinical intoxication and proteomics]. PMID- 17133979 TI - [Clinical toxicology and metabonomics]. PMID- 17133980 TI - [Post-marketing surveillance of acetylcysteine oral solution 17.6% "SENJU" for the antidote to acetaminophen overdose--use--results surveillance]. AB - Use-result surveillance was conducted to investigate the safety and efficacy of Acetylcysteine Oral Solution 17.6 % "SENJU" having the indication for the antidote to acetaminophen (Paracetamol) overdose. Ninety six cases (patients) were collected for the safety evaluation, and 13 cases (incidence was 13.5 %) showed 29 adverse drug reactions as follows: 4 cases of nausea; 3 cases of vomiting; 2 cases each of liver dysfunction, headache, abdominal pain, diarrhea, blood bilirubin increased; and one case each of CK increased, anaemia, prothrombin time prolonged, gamma-glutamyltransferase increased, LDH increased, body temperature increased, proteinuria, blood potassium decreased, thrombocytopenia, platelet count increased, white blood cell decreased, and blood amylase increased. One case of severe liver dysfunction which was ameliorated later was found. Neither case showing transitional chronic liver dysfunction, nor case of death was observed. Patient background analysis showed that 79.2% of the total patients was female, and that 28.1% was patients with mental disease. Gastrolavage, active charcoal administration, and extracorporeal removal of toxins were performed in cases of 71.9%, 50.0% and 7.3%, respectively. Those concomitant treatments, however, showed no influence for the incidence of adverse drug reaction or the drug effectiveness. Blood acetaminophen assay was performed in only 43.8% of the total cases. This rate indicates that the medical treatment procedure needs more consideration on the clinical standard for the antidote to acetaminophen overdose and on its practical application. PMID- 17133981 TI - Severe acute pancreatitis caused by organophosphate poisoning. AB - Organophosphate intoxication may be caused pancreatitis, but the role of diagnostic imaging for pancreatitis in these patients has not been well defined. We recently encountered a patient with organophosphate poisoning showing hyperamylasemia who was proven to have severe acute pancreatitis by CT findings. The patient was a 69-year-old woman who presented to a local hospital with disturbance of consciousness. She was initially treated for cerebral infarction, but an extremely low level of ChE was noted on Day 3. The patient was then referred to our institution. Examination of the abdomen revealed weak intestinal peristalsis, blood chemistry showed an increased level of serum amylase, and the urinary organophosphate test was positive. Based on the findings obtained by abdominal CT scanning, severe acute pancreatitis was diagnosed. Clouding of her consciousness resolved on day 21, but a pancreatic pseudocyst was detected on day 41. PMID- 17133982 TI - [Case clinically diagnosed as depressive state after being stung by a lumpfish (Inimicus japonicus)]. AB - We experienced and report a case where the patient was clinically diagnosed as depressive state which developed after being stung by a lumpfish-a kind of Japanese stonefish (Inimicus japonicus). Stonefish venom causes various symptoms ranging from local swelling with pain to general disturbances such as respiratory and heart failure with marked hypotension, cardiac perturbation, and neurologic damage including general seizure and coma. In the current case, the patient complained of local swelling with pain in the early stage, but subsequently he developed depressive state, and finally he began to have suicidal idea. When a patient is encountered who expresses severe depressive symptoms with suicidal idea, we hope that the patient can be examined by a psychiatrist, since the patient may have a serious accident or commit suicide during the process of the disease. On the other hand, it is easy to miss such depressive patients in cases where the depressive state appears after the appearance of toxic symptoms, and this is especially true in cases where the patient seems to be recovering naturally. In conclusion, we hope that medical institutions cooperate in analyzing the pathology of this toxicosis, since each institution rarely encounters such depressive patients. PMID- 17133983 TI - [Case of amoxapine-induced refractory status epilepticus which was successfully treated with propofol]. PMID- 17133984 TI - [Analytical research of cadmium in metal accessories]. PMID- 17133985 TI - [Proposal of an analytical pathway for the treatment of poisonings--6- application of blood concentration to the evaluation of toxicity (2): tools for diagnosis and therapy]. PMID- 17133986 TI - [Health hazard cases caused by powder fire extinguishers reported to Japan Poison Information Center]. PMID- 17133987 TI - [Sex differentiation in the brain]. PMID- 17133988 TI - [Toxicological analysis of a case of Datura stramonium poisoning]. AB - We encountered a patient in a restless excitable state after eating boiled jimson weed grown in the patient's garden. The patient mistook the weed for Angelica keiskei. Pupillary dilation (7/7mm), weak light reflex, body temperature of 37 degrees C, respiratory frequency of 19/min, blood pressure of 138/88 mmHg, pulse rate of 108/min, and hot feeling were observed. No abnormalities nor special findings were detected by general examination of the peripheral blood, biochemical examination of the blood, general examination of the urine, or electrocardiography. Atropine and scopolamine, which are tropane alkaloids, were detected by the GC/MS. The retention time of atropine-TMS was 17.0 min, and the mass spectra were m/z 124, 82, and 140. The retention time of scopolamine-TMS was 17.7 min, and the mass spectra were m/z 138, 108, 154 and 375. At the time of consultation, the serum concentrations of atropine and scopolamine were 31.3 ng/ml, and 30.6 ng/ml, respectively, and decreased to 6.7 ng/ml and 8.5 ng/ml, respectively, after 2 hours. The patient underwent injection of activated carbon after gastrolavage with 2,000 ml warm water, and neostigmine was administered. The patient awoke the following morning, and was discharged with mild pupillary dilation 2 days after poisoning. PMID- 17133989 TI - [Controversies in the determination of serum albumin concentration in chronic liver diseases]. AB - OBJECTIVE: It is known that there is quantitative alteration in serum albumin in the patients with chronic liver diseases. To clarity the quantitative alteration of serum albumin, serum albumin levels were measured by the Bromcresol green (BCG) method, the modified Bromcresol purple (BCP) method and immunoassay. METHODS: We enrolled 313 patients with chronic hepatitis, 74 cirrhotic patients and 36 healthy subjects. Serum albumin levels were measured by immunoassay, the BCG method and the modified BCP methods. RESULTS: The measurement of immunoassay showed a significant correlation in the measurement of the modified BCP method in the patients with chronic liver diseases. With the progress of chronic liver diseases, the serum albumin levels by the BCG method and electrophoresis were higher than the levels by the modified BCP method. In other hands, it was 0.07 g/dl higher level in chronic hepatitis and 0.18 g/dl higher in liver cirrhosis. And the serum albumin levels by the BCG method and electrophoresis were higher than the levels by the new BCP method in the patients with leg edema and ascites. In the malnourished patients Subjective global assessment (SGA-B), the serum albumin levels by the BCG method and electrophoresis were higher than the levels by the modified BCP method. CONCLUSIONS: Serum albumin levels were different by the method for measurements. The clinical implications of quantitative changes in albumin should be investigated in consideration of the microheterogeneity of albumin. The modified BCP method is superior method to evaluate the serum albumin levels. PMID- 17133990 TI - [Progress of the Laboratory Consultation Office in Kyoto Prefectural University Hospital]. AB - In an attempt to assist the clinical physicians at the outpatient clinics in Kyoto Prefectural University Hospital, we opened the laboratory consultation office in January 2005. This office is staffed by one medical technologist from 9 am to 4 pm on weekdays. The technologist shows the laboratory examination result of respective patient, and then explains the significance of reference interval using the personal computer. Additional explanation such about the mechanism of abnormal result or the general interpretation of diagnostic tests is also performed according to the patients' demands. Whereas, as the major premise, the consulting technologist never comments to the patients on the clinical relevance between their laboratory results and their own disease courses. Until March 2006, 1,430 outpatients have visited the office in total, resulting in 6.1 patients/day as mean value. Almost all of the visitors (97.9%) wanted to listen to the meaning of liver function test, which was followed by renal function test (95.3%). Consulting time required for respective patient has been quite variable from 10 to 120 minutes (30 minutes/patient as mean). The questionnaire survey after consultation revealed that all of the visitors felt satisfactory for consultation, and wanted to revisit to our laboratory consultation office. We believe that the laboratory consultation office contributes as a new medical service of our hospital to outpatients. PMID- 17133991 TI - [The path I have studied in clinical laboratory medicine for thirty years and ten months]. PMID- 17133992 TI - [Introduction of Department of Diagnostic Medicine, Graduate School of Medical Sciences, Kumamoto University]. AB - Pathologic informatics is an analysis of mechanism of pathogenesis using various medical information. Biologic reactions by many reactants are studied on molecules, cells, organs, and individuals by using various methods. Themes of our laboratory are as follows: 1) to analyze of the pathogenesis of various diseases by using novel diagnostic methods, 2) to elucidate amyloid formation mechanism in amyloidosis and to develop novel therapies, 3) to develop the methods for tests and diagnosis for DIC, 4) to study on activation mechanism of ribosomal proteins in the tissues by the blood cell function, 5) to study on the expression and suppression of neogenesis of blood vessels by physical invasion in the organ failure, and 6) to develop the diagnostic methods for autonomic dysfunction and to analyze the pathogenesis. PMID- 17133993 TI - [Feeding-related disorders in medicine, with special reference to cancer anorexia cachexia syndrome]. AB - Cachexia is among the most debilitating and life-threatening aspects of cancer. Associated with anorexia, fat and muscle tissue wasting, psychological distress, and a lower quality of life, cachexia arises from a complex interaction between the cancer and the host. This process results from a failure of the adaptive feeding response seen in simple starvation and includes cytokine production, release of lipid-mobilizing and proteolysis-inducing factors, and alterations in intermediary metabolism. Cytokines play a pivotal role in long-term inhibition of feeding by mimicking the hypothalamic effect of excessive negative feedback signaling from leptin, a hormone secreted by adipose tissue, which is an integral component of the homeostatic loop of body weight regulation. The two major options for pharmacological therapy have been either progestational agents or corticosteroids. However, knowledge of the mechanisms of cancer anorexia-cachexia syndrome continues to lead to effective therapeutic interventions for several aspects of the syndrome. These include antiserotonergic drugs, gastroprokinetic agents, branched-chain amino acids, eicosapentanoic acid, cannabinoids, melatonin, and thalidomide, all of which act on the feeding-regulatory circuitry to increase appetite and inhibit tumor-derived catabolic factors to antagonize tissue wasting and/or host cytokine release. The outcomes of drug studies in cancer cachexia should focus on the symptomatic and quality-of-life advantages rather than simply on nutritional end points, since the survival of cachexia cancer patients may be limited to weeks or months due to the incurable nature of the underlying malignancy. As weight loss shortens the survival time of cancer patients and decreases their performance status, effective therapy would extend patient survival and improve quality of life. PMID- 17133994 TI - [RNA amplification from low numbers of bacteria in human blood to solve low sensitivity problem of conventional PCR amplification]. AB - Blood culture from a febrile patient is often unsuccessful when a number of bacteria is lower than 10 CFU/ml in circulation. Theoretically, PCR amplification of bacteria in this low bacterial level in human blood is beyond the sensitivity of PCR method. RNA based Amplification (NASBA) method is more sensitive than PCR base assay because living bacteria carries several hundred to thousand RNA molecules in a cell. Increasing evidences also revealed another problem that human blood stream is often contaminated with bacterial genome. This often cause false positive results and bring difficulty of the interpretation to determine causative agent. Direct DNA amplification should be limited to severe causative agent such as Haemophilus influenzae, Streptococcus pneumoniae, Neisseria meningitides, and etc. and also applied only for slow growing and fastidious pathogens such as mycoplasmae, chlamydiae, bartonellae, mycobacteria, and etc. Most cases blood culture should be carried out first and genetic amplification should be selected as a next step to detect pathogens from a blood culture bottle, because in these cases, patients are not in life threatening stages. These attempts also decrease the chance to detect circulating bacterial DNA fragment in blood stream. PMID- 17133995 TI - [Interpretation of the results of blood culture and interventional role by clinical laboratory physicians]. AB - Blood culture has long been recognized as the gold standard for the definitive diagnosis of bacterial and fungal infections. However, fewer blood cultures have been tested and their results have not been fully used in Japan. Clinical laboratory physicians should play an interventional role, such as recommending blood culture tests in patients with infectious disease or fever of unknown origin. In our hospital, clinical laboratory physicians act as on-call consultants. The yearly number of consultations is between 500 and 700, and consultations concerning infectious disease have increased up to 40% in the past 5 years. As a result, the number of blood cultures and the percentage of 2-set blood collections have increased in order to increase the positivity rate and determine whether the results obtained were contaminated. However, physicians sometimes misunderstand the results of blood culture, and they assume that the identified organism was causative, or that sepsis did not exist if the culture is negative. Clinical laboratory physicians should act as consultants more frequently, concerning the interpretation of blood culture results, and the choice of antimicrobial agents, because the inappropriate use of antimicrobial agents leads to higher mortality and higher medical costs. Finally, collaboration between clinical laboratory physicians and co-medical staff such as the infection control team, nurses and pharmacists is necessary. PMID- 17133996 TI - [Guidelines for intravenous thrombolytic therapy in acute ischemic stroke with special reference to those by the Ad Hoc Committee of the Japan Stroke Society]. PMID- 17133997 TI - [Diagnostic imaging of acute ischemic stroke]. PMID- 17133998 TI - [Subjects in the acute thrombolytic therapy by intravenous administration of rt PA]. PMID- 17133999 TI - [Neuroendovascular therapy for acute stroke]. PMID- 17134000 TI - [Paradoxical brain embolism]. PMID- 17134002 TI - [Newly published Neurological Clinical Pathological Conference]. PMID- 17134001 TI - [Antiphospholipid syndrome]. PMID- 17134003 TI - [Cerebral arterial dissection]. PMID- 17134004 TI - [Cryptogenic stroke--vascular malformation-clinical features]. PMID- 17134005 TI - [Study of movement disorder with network analysis]. PMID- 17134006 TI - [Pathogenesis of myasthenia gravis]. PMID- 17134007 TI - [Strategy for management of carotid arterial stenosis: analysis of 293 consecutive patients in a Japanese cerebrovascular center]. AB - Carotid arterial stenosis is a major risk factor for ischemic stroke and is increasing in Japan as the life-style has been westernized. The purpose of this study was to clarify the detailed process of diagnosis and treatment of patients with carotid arterial stenosis. Of the consecutive 1,889 hospitalized patients in our cerebrovascular center during 2001 and 2003, 293 patients had carotid stenosis 50% or more in diameter by the NASCET method; 82 patients were hospitalized during the acute stage of ischemic stroke and 211 patients with or without past history of ischemic stroke were admitted in the chronic stage. Among acute ischemic stroke patients, 62 patients (76%) had mild neurological symptoms of NIH Stroke Scale score < or = 4 on admission. As the initial treatment during the acute phase, all patients underwent antithrombotic medication; 33 of them underwent carotid endarterectomy (CEA) or carotid arterial stenting (CAS) in the chronic stage. Of 211 chronic patients, 123 (58%) did not have a history of symptomatic ischemic stroke, and instead had nonspecific symptoms, including carotid bruit, headache, and vertigo, or were diagnosed as having carotid artery stenosis by examinations of preoperative screenings. One hundred and thirty-five chronic patients underwent CEA/CAS and all the others except for a patient with serious gastrointestinal bleeding underwent anti-thrombotic medication. Statin treatment was chosen for 59 acute patients and 66 chronic patients. Because many patients with carotid arterial stenosis had mild symptoms during the acute phase or did not have ischemic episodes, we might overlook carotid lesions unless we performed screening examinations using ultrasound or magnetic resonance angiography. PMID- 17134008 TI - [Case of gustatory disturbance caused by pontine infarction]. AB - We reported a 72-year-old man presented with gustatory disturbance. He lost bitter, salty, sour and sweet sensation on the left half of the tongue. No other neurologic signs were observed. Brain MRI showed the cerebral infarction in the left middle cerebellar peduncle. The lesion was suspected of affecting the gustatory tract running from the nucleus solitalius to the pontine taste area in the upper pons. The gustatory tract is generally recognized to be located medially from the medial lemniscus and the reticular formation. Our case suggests that the tract would be located laterally from the medial lemniscus and the reticular formation. PMID- 17134009 TI - [Adult case of acute encephalopathy associated with bilateral thalamic lesions and peripheral neuropathy]. AB - A 76-year-old woman developed fever and consciousness disturbance. The next day, she became delirious and was brought to our hospital. On arrival, she was unconscious and showed hypopnea and hypotension. She was immediately intubated and placed on a respirator. CSF protein was 65.8 mg/dl with 1 cell/microl, and no oligoclonal bands were present. An electroencephalogram showed diffuse theta background activity without epileptic discharges. A nerve conduction study showed damaged motor and sensory peripheral nerve functions in the upper and lower limbs. The neurological findings showed no improvement after methylprednisolone pulse therapy and administration of intravenous immunoglobulin. Magnetic resonance imaging of the brain, including diffusion-weighted images showed bilateral symmetric lesions in the thalamus, globus pallidus and pontine tegmentum. These radiologic findings are not typically, but are similar to those of acute necrotizing encephalopathy (ANE) of childhood as proposed by Mizuguchi et al. After 10 months, brain MRI showed bilateral brain atrophy and a reduction of the abnormal thalamic lesions. There are very few reports of adult cases of ANE, in which, pathologically, local breakdown of the blood-brain-barrier causes acute edema and necrosis involving both gray and white matter. ANE is thought a proinflammatory cytokine-related disease. In our case, the concentrations of some cytokines (IL-6, IL-10) were elevated in serum and cerebrospinal fluid, which might suggest a relationship with them and local breakdown of the blood-brain barrier in the thalamus. PMID- 17134010 TI - [Case of an adult presenting with cerebral atrophy in MRI in the course of encephalitis and later improvement in MRI findings]. PMID- 17134011 TI - [Experimental approach to murder by aconite poisoning from the viewpoint of medicolegal toxicology]. AB - An autopsy case performed by the author in 1986 had been gradually revealed to be a murder using aconite poisons. The puffer fish toxin was certified afterwards to be co-administered together with aconite alkaloids in this case. In order to investigate this murder case, animal experiments were done using mice to clarify the metabolism of aconitine and tetrodotoxin, and to examine the influences of tetrodotoxin on aconite poisoning. We also examined biological effects under the chronic intoxication of aconitine, and the elimination and degradation of aconitine in dead body. For this purpose we have developed technical methods using GC/MS and LC/MS for the quantification of these toxins in biological materials. PMID- 17134012 TI - [Molecular approaches to the Rh blood group system]. AB - The Rh system clinically is one of the important blood groups. The major Rh antigens are RhD, RhC/c, and RhE/e, which are carried by two integral membrane polypeptides consisted of 416 amino acids. These polypeptides are encoded by two closely related genes, RHD and RHCE. Both RH genes are composed of ten exons. It is thought that multiple recombinations, nucleotide substitutions, large nucleotide gaps (due to Alu sequence), and high level of the homology between the RHD and RHCE genes are the important factors in the formation and evolution of these genes. The RHD gene is deleted in most white individuals who lack the RhD antigen, while 12% of Japanese individuals have an RHD gene. Molecular analyses have elucidated the background of various Rh-related variants; D--, partial D, weak D, and Rhnull. The Rhnull phenotype is divided into the most common type by the Rhnull regulator gene and second type by the amorph gene that arose by homozygosity of a silent allele at the RH locus. The RhAG glycoprotein has been regarded as a most critical Rhnull gene of the reglurator type and a critical co expressing factor of the Rh polypetides on red blood cells. Studies on the autoantibodies against red blood cells in aoutoimmune hemolytic anemia have suggested that the public epitopes of autoantigens exist on the Rh polypeptides. PMID- 17134013 TI - [Medico-legal autopsy case of an infant suffering anaphylactic shock during dental treatment. Potential hazards in the use of a rubber-dam-sheet for infants]. AB - We report a medico-legal autopsy of a 4-year-old girl without any history of allergies, who died following anaphylactic shock due to local anesthesia with lidocaine during dental treatment. The shock symptoms were overlooked due to a rubber-dam-sheet covering her face. The autopsy revealed laryngeal edema, secretory fluid filling the trachea, severe pulmonary congestion and edema. An immunohistochemical investigation showed numerous mast cells releasing histamine in lungs, and the fluids in the pulmonary alveoli and veins also showed positive staining. In addition, the plasma concentration of histamine in the heart blood showed a high value. From these findings, the cause of death was determined to be respiratory failure due to anaphylactic shock induced by lidocaine. In this case, her death was considered to be due to medical malpractice; adequate life support had not been performed, because anaphylactic shock was overlooked until the rubber-dam-sheet removed. In addition, due to two previous autopsy case reports that described infantile fatalities involved with the misusage of a rubber-dam sheet for the patients under local anesthesia, we therefore recommend immediate improvements in rubber-dam-sheets and/or better applications of them in pediatric dentistry. PMID- 17134014 TI - [Natural death in adults: part 2. Autopsy]. PMID- 17134015 TI - [Injury of the trunk]. PMID- 17134016 TI - Evaluation of alveolar clearance by Tc-99m DTPA radioaerosol inhalation scintigraphy in welders. AB - OBJECTIVES: The welding process produces metal fumes and gases which may affect respiratory health. Technetium-99m diethylenetriaminepentaacetic acid (Tc-99m DTPA) dynamic lung scanning is an easy, noninvasive method to assess disorders of alveolar-capillary barrier permeability secondary to epithelial damage. We aimed to investigate the alveolar clearance by Tc-99m DTPA radioaerosol inhalation scintigraphy in welders, to assess additive effects of exposure to welding fumes and cigarette smoking on clearance rate of alveolar epithelium and to determine the correlation between Tc-99m DTPA aerosol lung scintigraphy and spirometric measurements. METHODS: Nine nonsmoking welders, 9 smoking welders, and a control group of 6 nonsmokers and 6 smokers were accepted to the study. Tc-99m DTPA radioaerosol inhalation scintigraphy was performed in all subjects. Clearance half time (Tl/2) was calculated by placing a monoexponential fit on the curves. Penetration index (PI) was also calculated on the first minute image. Pulmonary function tests of welders and control group were compared. RESULTS: The mean T1/2 values of Tc-99m DTPA of the nonsmoking welders were significantly higher than those of the nonsmoking control group (82.1+/-24.3 min and 48.1+/-9.7 min, respectively; p = 0.003). The mean T1/2 values of Tc-99m DTPA of the smoking welders were higher than those of the smoking control group (53.3+/-24.5 min and 44.5+/-9.7 min, respectively; p = 0.510). PI of the nonsmoking welders was significantly higher than that of the nonsmoking control group (0.46+/-0.38 and 0.39+/-0.46 respectively; p = 0.004). PI of the smoking welders was significantly higher than that of smoking control group (0.43+/-0.38 and 0.37+/-0.45, respectively; p = 0.019). There was a negative correlation between T1/2 value and FEV1% (r = -0.468, p = 0.016), FVC% (r = -0.442, p = 0.024) and FEF25-75% (r = 0.391, p = 0.048) in the welders and control group. No statistically significant differences were found in the values of the standard pulmonary function tests of any of the subjects. CONCLUSIONS: Welding seems to decrease alveolar clearance which causes an increase in the penetration index. This was considered to be due to fibrotic changes and increased number of alveolar macrophages induced by welding fumes. PMID- 17134017 TI - Effect of linearization correction on statistical parametric mapping (SPM): a 99mTc-HMPAO brain perfusion SPECT study in mild Alzheimer's disease. AB - OBJECTIVE: Statistical parametric mapping (SPM) was employed to investigate the regional decline in cerebral blood flow (rCBF) as measured by 99mTc-hexamethyl propylene amine oxime (HMPAO) single photon emission computed tomography (SPECT) in mild Alzheimer's disease (AD). However, the role of the post reconstruction image processing on the interpretation of SPM, which detects rCBF pattern, has not been precisely studied. We performed 99mTc-HMPAO SPECT in mild AD patients and analyzed the effect of linearization correction for washout of the tracer on the detectability of abnormal perfusion. METHODS: Eleven mild AD (NINCDS-ADRDA, male/female, 5/6; mean+/-SD age, 70.6+/-6.2 years; mean+/-SD mini-mental state examination score, 23.9+/-3.41; clinical dementia rating score, 1) and eleven normal control subjects (male/female, 4/7; mean+/-SD age, 66.8+/-8.4 years) were enrolled in this study. 99mTc-HMPAO SPECT was performed with a four-head rotating gamma camera. We employed linearization uncorrected (LU) and linearization corrected (LC) images for the patients and controls. The pattern of hypoperfusion in mild AD on LU and LC images was detected by SPM99 applying the same image standardization and analytical parameters. A statistical inter image-group analysis (LU vs. LC) was also performed. RESULTS: Clear differences were observed between the interpretation of SPM with LU and LC images. Significant hypoperfusion in mild AD was found on the LU images in the left posterior cingulate gyrus, right precuneus, left hippocampus, left uncus, and left superior temporal gyrus (cluster level, corrected p < 0.005). With the LC images, significant hypoperfusion in AD was found only in the bilateral posterior cingulate gyrus and left precuneus (cluster level, corrected p < 0.005). A pattern of greater rCBF distribution at the high flow cortices and low flow cortices was observed on LC and LU images, respectively, in the case of both controls and mild AD patients. CONCLUSION: Hippocampal hypoperfusion could be detected by means of SPM in the LU images but not in the LC images. The results of SPM may vary in 99mTc-HMPAO SPECT with or without linearization correction, which should be carefully evaluated when interpreting the pattern of rCBF changes in mild Alzheimer's disease. PMID- 17134018 TI - Evaluation of right and left ventricular function by quantitative blood-pool SPECT (QBS): comparison with conventional methods and quantitative gated SPECT (QGS). AB - Though quantitative ECG-gated blood-pool SPECT (QBS) has become a popular tool in research settings, more verification is necessary for its utilization in clinical medicine. To evaluate the reliability of the measurements of left and right ventricular functions with QBS, we performed QBS, as well as first-pass pool (FPP) and ECG-gated blood-pool (GBP) studies on planar images in 41 patients and 8 healthy volunteers. Quantitative ECG-gated myocardial perfusion SPECT (QGS) was also performed in 30 of 49 subjects. First, we assessed the reproducibility of the measurements of left and right ventricular ejection fraction (LVEF, RVEF) and left and right ventricular end-diastolic volume (LVEDV, RVEDV) with QBS. Second, LVEF and RVEF obtained from QBS were compared with those from FPP and GBP, respectively. Third, LVEF and LVEDV obtained from QBS were compared with those from QGS, respectively. The intra- and inter-observer reproducibilities were excellent for LVEF, LVEDV, RVEF and RVEDV measured with QBS (r = 0.88 to 0.96, p < 0.01), while the biases in the measurements of RVEF and RVEDV were relatively large. LVEF obtained from QBS correlated significantly with those from FPP and GBP, while RVEF from QBS did not. LVEF and LVEDV obtained from QBS were significantly correlated with those from QGS, but the regression lines were not close to the lines of identity. In conclusion, the measurements of LVEF and LVEDV with QBS have good reproducibility and are useful clinically, while those of RVEF and RVEDV are less useful compared with LVEF and LVEDV. The algorithm of QBS for the measurements of RVEF and RVEDV remains to be improved. PMID- 17134019 TI - Comparison of 18F-fluoromethylcholine and 2-deoxy-D-glucose in the distribution of tumor and inflammation. AB - PURPOSE: The distribution characteristics of 18F-fluoromethylcholine (18F choline) in tumor and inflammatory tissue were compared with those of 14C or 3H-2 deoxyglucose (2DG) as a substitute for fluorodeoxyglucose (FDG). METHODS: A solid tumor model of AH 109A in the back of Donryu rats and an aseptic inflammation model of turpentine oil injection subcutaneously in rats were used for experiments. Tissue distribution was examined at 5, 30 and 60 min after injection of a mixture of 18F-choline and 3H-2DG. Double-tracer high-resolution autoradiographs (ARGs) of tumor and inflammation were obtained using 18F-choline and 14C-2DG. Whole body (WB) ARG was performed with 18F-choline. RESULTS: Tumor uptake of 18F-choline reached a peak at 30 min, when the tumor to blood ratio was 5.1. Both tumor and inflammation uptake of 2DG were higher than those of 18F choline. 18F-choline uptake by inflammation was lower than that by tumor. The tumor to brain uptake ratio was 5.7 with 18F-choline and 1.2 with 2DG. In the ARG of inflammation, linear or ring-like structures of 2DG uptake were observed in the wall of the abscess, but were not identified with 18F-choline. Photomicrography showed that the uptake was limited to granulocytes, macrophages and fibroblasts, consistent with sub-acute or chronic inflammation. CONCLUSION: 18F-choline uptake by inflammation was lower than that of 2DG in the tissue distribution study, and 18F-choline uptake by abscess wall was significantly lower than that of 2DG in the autoradiography study. Our results may suggest the feasibility of 18F-choline-PET imaging for the differential diagnosis of cancer and chronic inflammation in lung and brain. PMID- 17134020 TI - Pharmacokinetics and biodistribution of a small radioiodine labeled nerve growth factor fragment. AB - Nerve growth factor (NGF) exerts various actions on neuronal and non-neuronal tissues and has potential therapeutic utility, but difficulties in using the whole protein have stimulated interest in small NGF fragments. We radioiodinated a small cyclic peptide derived from NGF using the Bolton-Hunter method [125I-C(92 96)], and confirmed binding to high affinity NGF receptors by cross-linkage analysis. Pharmacokinetic characteristics in intravenously injected mice were T 1/2 alpha 5.2 min, T 1/2beta 121.3 min, clearance 11.8+/-0.5 ml/min, and volume of distribution 69.7+/-4.6 ml. Dose-proportionate increases in areas-under-curve and peak-concentrations indicated linear pharmacokinetics. Biodistribution data revealed that clinically relevant doses allowed C(92-96) accumulation sufficient to elicit biological responses in receptor expressing organs including the lungs, liver, spleen, and pancreas. PMID- 17134021 TI - Effects of diazepam on 125I-iomazenil-benzodiazepine receptor binding and epileptic seizures in the El mouse. AB - OBJECTIVE: To investigate changes in free benzodiazepine receptor density in response to repeated, long-term administration of diazepam in epilepsy, we assessed 125I-iomazenil (125I-IMZ) binding in a mouse model. METHODS: El mice were divided into two groups of 12 mice each which received either no diazepam (E1(D[-]) group) or 2 mg/kg of diazepam per week (El(D[+]) group). Nine ddY mice were used as a control. Once each week from the age of 5 to 19 weeks, the El mice received stimulation to produce epileptic seizures 20 minutes after receiving intraperitoneal injections. At 20 weeks of age, a total dose of 0.37 MBq of 125I IMZ was injected in all mice and their brains were rapidly removed 3 hours later. The incidence of epileptic seizures at the age of 19 weeks and the autoradiograms of the brain were compared. RESULTS: The incidence of epileptic seizures in response to weekly stimulation was significantly lower in the E1(D[+]) group than in the E1(D[-]) group (p < 0.001). The percent injected doses of 125I-IMZ per gram of tissue in the cortex, hippocampus and amygdala were significantly lower in the E1(D[+]) group than in the E1(D[-]) group (p < 0.05). CONCLUSION: The results suggest that diazepam binds competitively to 125I-IMZ as an agonist to free benzodiazepine receptor sites in the cortex, hippocampus and amygdala and shows anticonvulsant effect in E1 mice. PMID- 17134022 TI - The clinical impact of 18F-FDG PET in papillary thyroid carcinoma with a negative 131I whole body scan: a single-center study of 108 patients. AB - OBJECTIVE: To assess whether FDG PET could localize the recurrent or metastatic lesions in papillary thyroid cancer patients with negative radioiodine scan. METHODS: Whole body PET was performed after injecting 370-555 MBq of 18F-FDG in 108 patients, who were suspected of having recurrence or metastasis and whose 131I whole body scans were negative. Recurrence or metastasis occurred in 63 patients by pathology or clinical assessment, whereas 45 patients remained in remission. RESULTS: FDG PET revealed recurrence or metastases in 59 patients (sensitivity 93.7%), whereas thyroglobulin (Tg) levels were elevated in 41 (sensitivity 65.1%). In 35 of 45 patients in remission, FDG PET was negative (specificity 77.8%). When patients positive for antithyroglobulin antibody were excluded, the sensitivity and specificity of serum Tg became 84.8% and 46.9%, respectively. Compared to Tg measurement, FDG PET detected more metastatic lesions in cervical lymph nodes. Of 40 patients with a negative radioiodine scan showing diffuse hepatic uptake, metastases occurred in 23 patients and remission in 17. FDG PET showed 100% sensitivity and 76.5% specificity in the detection of recurrence in these 40 patients. CONCLUSION: FDG PET is useful for localizing recurrent or metastatic lesions in 131I scan-negative thyroid cancer patients. In particular, it is superior to serum Tg measurement for identifying metastases to cervical lymph nodes. We recommend its use in cases of negative radioiodine scan with diffuse hepatic uptake. PMID- 17134023 TI - 18F-FDG PET/CT findings of a right subphrenic foreign-body granuloma. AB - We report a case of an 85-year-old woman with a foreign-body granuloma which accumulated 18F-fluorodeoxyglucose (FDG). Unenhanced computed tomography showed a hyperdense mass with a hypodense rim in the right subphrenic space. FDG PET/CT images showed intense FDG uptake in the hypodense rim and little FDG uptake in the center of the mass, showing a ring-shaped appearance. The fusion imaging of FDG PET/CT represented the metabolic features of the foreign-body granuloma. When a ring-shaped FDG uptake is noted in the abdomen of a patient with a history of abdominal surgery, a foreign-body granuloma should be included in the differential diagnosis. PMID- 17134024 TI - Visualization of gallbladder with In-111 labeled octreotide in post prandial state. AB - Somatostatin receptor scintigraphy is widely used in the management of neuroendocrine tumors. Somatostatin receptors are present in both neoplastic and normal tissues, which may lead to misinterpretation of the scans. Here, a patient with lung carcinoid imaged with In-111 octreotide is presented. Imaging was performed 4 and 24 hours after an intravenous injection of 185 MBq In-111 octreotide in the post prandial state. Whole body and SPECT images showed accumulation of radioactivity in the gallbladder. Imaging was repeated after fatty meal ingestion to differentiate abnormal activity and physiological uptake in the gallbladder. The abdominal SPECT studies at 28 hours revealed no uptake in the gallbladder, and the scintigraphic study was reported as normal so further excessive diagnostic procedures were prevented. Gallbladder can be visualized on somatostatin receptor scintigraphy even in the post prandial state. Delayed images after fatty meal administration are important for differential diagnosis. PMID- 17134025 TI - Accumulation of Tc-99m-HMPAO-labeled leukocytes in cutaneous lesions of Henoch Schonlein purpura. AB - A 70-year-old man with complicated cruris fracture, treated with an arthrodesis and skin transplantation, complained about persistent pain of the right ankle and skin rashes. Bone and leukocyte scintigraphy were performed because osteomyelitis was suspected. Bone scintigraphy showed abnormal uptake in the ankle. Leukocyte scintigraphy showed only multiple hot spots in the soft tissue. A cutaneous biopsy specimen revealed Henoch-Schonlein vasculitis. PMID- 17134026 TI - Tc-99m nanocolloid scintigraphic imaging of intracranial meningeal extramedullary hematopoiesis in a patient with idiopathic myelofibrosis. AB - Meningeal extramedullary hematopoiesis (EMH) is a rare finding in idiopathic myelofibrosis. Intracranial EMH is typically asymptomatic and sites are usually found by chance. Diagnosis of EMH is difficult, based on clinical circumstances and the use of different diagnostic imaging modalities, such as CT, MRI or radionuclide imaging. We present a case with intracranial medullary hematopoiesis due to idiopathic myelofibrosis diagnosed with Tc-99m nanocolloid scintigraphy. Cranium SPECT images that were performed with Tc-99m nanocolloid showed increased radiotracer uptake in the bilateral parietal, bilateral frontal and left occipital bones and especially in falx cerebri of sinus sagittalis superior. In Tc-99m MDP bone scintigraphy, increased osteoblastic activity in the left frontal and parietal bones, in shoulders, knee and ankle joints, and in both metatarsal bones were seen. After gadodiamid injection, Tl weighted MRI showed diffuse contrast increased in the meningeal areas surrounding the brain. A biopsy of the mass revealed extramedullary hematopoiesis composed of erythroblasts, mature and immature myeloid cells, and megakaryocytes. It was deduced that these described foci of EMH. PMID- 17134027 TI - A feasibility study of [11C]SA4503-PET for evaluating sigmal receptor occupancy by neuroleptics: the binding of haloperidol to sigma1 and dopamine D2-like receptors. AB - We investigated feasibility of positron emission tomography (PET) with [11C]SA4503 for evaluating the sigma1 receptor occupancy rate by neuroleptics. Haloperidol, which is well known to bind dopamine D2-like receptor (D2R) as well as to be a representative non-selective antagonist for sigma1 receptor (sigma1R), was selected as a model drug. Three healthy male subjects underwent 60-min [11C]raclopride-PET and 90-min [11C]SA4503-PET scans successively at a 120-min interval twice in a day for baseline measurement and on another day for haloperidol-loading measurement 16 hours after peroral administration of 3 mg of haloperidol. Binding potential (BP) of [11C]raclopride and [11C]SA4503 was quantitatively evaluated and the sigma1R and D2R occupancy rates were determined. D2R occupancy rates by haloperidol were 64% and 62% in the caudate and putamen, respectively, 16 h after the administration, while sigma1R occupancy rates were approximately 80% in all seven regions investigated including the caudate, putamen and cerebellum 18 h after the administration, suggesting that the sigma1R receptor occupancy rate by haloperidol was slightly larger than the D2R receptor occupancy rate. We concluded that [11C]SA4503-PET can be used for evaluating the sigma1R occupancy rates by neuroleptics or other drugs. PMID- 17134028 TI - Radiopharmaceutical management of 90Y/111In labeled antibodies: shielding and quantification during preparation and administration. AB - BACKGROUND: The combined application of potent beta-emitting isotopes for therapy with remitting isotopes for scintigraphy requires a profound regimen concerning team member safety and radionuclide quantification. METHODS: We have developed materials and methods for a proper and easy manipulation of 90Y during preparation and administration of 90Y/111In pharmaceuticals used for radioimmunotherapy. RESULTS: The efficacy of the shielding measures is documented. Protocols for the calibration of gamma-dose calibrators with respect to 90Y are extended to the assessment of quench-corrected liquid scintillation counting of 90Y. The contribution of 90Y backscatter to 111 In counting is quantified. Newly developed shielding equipment allows an adequate administration of relatively large volumes (100 ml) of 90Y/111In labeled pharmaceuticals to patients. CONCLUSIONS: The procedures described combine pharmaceutical (Good Manufacturing Practice) and radiation safety requirements with an accurate logging of relevant data. PMID- 17134029 TI - [Molecular pathology and therapeutics in Alzheimer's disease: amyloid beta protein as a therapeutic target]. PMID- 17134030 TI - Geriatric patients presenting to the emergency department of a Japanese university hospital. AB - To evaluate the trend of elderly patients visiting the emergency department of a Japanese University Hospital, out patient-based records were reviewed of the emergency department of Kyushu University Hospital from 2000 to 2004. A total number of 7610 emergency patients visited the department during the five year period. The median (25%, 75%) of age was 32 (22, 56). Patients aged 65 years and over accounted for 16% of all attendances. All the patients were classified into 6 groups according to the diagnosis, (1) Respiratory, (2) Circulatory, (3) Central nervous system, (4) Abdominal, (5) Trauma, and (6) Others. The median age in each group was (1) 27 (15, 49), (2) 66 (53, 76), (3) 51 (27, 67), (4) 33 (22, 56), and (5) 26 (20, 46), respectively. There was a statistically significant difference observed, reciprocally except between (1) and (5) (P < 0.05). The patients showed statistically significant difference in the annual transition of the disease (P < 0.0001). In the elderly, the annual transition of the disease showed statistically significant decreases in Circulatory (P = 0.0015) and in Central nervous (P < 0.0001), and an increase in Abdominal (P < 0.0001), respectively. Death rate at the outpatient clinic in the elderly showed much higher than in the younger (P < 0.0001). Admission rate was also much higher in the elderly than in the younger (P < 0.0001). Elderly emergency patients have both internal and external intrinsic factors. They have to be treated carefully since their condition easily deteriorates. Provisions for the problems surrounding the elderly should be made as a nationwide effort. PMID- 17134031 TI - Short term intensive insulin therapy improves insulin secretion significantly in type 2 diabetic patients. AB - To investigate the effects of short-term (1 week) intensive insulin therapy, on glycemic control, insulin secretion, and insulin sensitivity in type 2 diabetic patients, an open prospective study was conducted in sixteen type 2 diabetic patients receiving diet therapy alone or treatment with oral hypoglycemic agents. Of the study subjects, 8 patients were treated with insulin, the remaining 8 patients served as the control group. The metabolic parameters were evaluated once before treatment and once during one of the following treatments : glycemic control as measured by 1,5-anhydro-D-glucitol (1,5-AG) and area under curve of glucose (AUCglucose), insulin secretion as measured by area under curve of daily serum insulin (AUCinsulin), and insulin sensitivity as measured by the K index of the insulin tolerance test (K(ITT)). Post-treatment plasma glucose (AUCglucose) and 1,5-AG levels in patients who had received intensive insulin therapy were comparable to those of the control group. A statistically significant increase in AUCinsulin occurred after intensive insulin therapy for just 1 week, while no change occurred in the control group. Insulin sensitivity (K(ITT)) did not improve significantly in patients treated with insulin or patients from the control group. These results indicate that intensive insulin therapy for 1 week improves insulin secretion remarkably but has little effect on insulin sensitivity in type 2 diabetic patients. Clinically, this suggests that intensive insulin therapy for one week might be one of the initial treatments of choice for such patients. PMID- 17134032 TI - [Novel approaches for fundus lesions in high myopia]. PMID- 17134033 TI - [Pathogenesis and treatment of myopic foveoschisis]. AB - Myopic foveoschisis (MF) can now be recognized as the result of recent progress in imaging diagnostic technology such as optical coherence tomography (OCT). MF predominantly occurs in highly myopic eyes with posterior staphyloma, and often induces visual loss and/or metamorphopsia. In order to better understand the detailed pathogenesis of MF, we observed 21 myopic eyes after vitrectomy using OCT. Retinal microfolds along the retinal vessels were observed in over 60% of the patients 6 months after surgery. Relative inflexibility of retinal arterioles/venules may be one of the causes for MF. We performed a pilot study (6 eyes of 5 patients) to treat MF by vitrectomy including vitreous cortex removal, internal limiting membrane peeling, and gas tamponade. In all 6 eyes the fovea was reattached and vision improved by more than two lines. This pilot study indicates that vitrectomy is an effective and safe treatment for MF. PMID- 17134034 TI - [Potential for neural regeneration in the adult mammalian retina]. AB - PURPOSE: It has long been believed that the retina of mature mammals is incapable of regeneration. However, here we show that Muller glia of adult mammals could be progenitor cells, and generate new retinal neurons. METHODS: N-methyl-D aspartate(NMDA) was injected into the vitreous chamber of adult rat (postnatal 6 7 weeks) eyes to induce neurotoxic injury. We injected bromo-deoxyuridine (BrdU) into the vitreous chamber and intraperitoneal space, and performed immunohistochemistry staining for BrdU and cell specific markers. To test whether exogenous growth factors stimulate proliferating cells, we injected retinoic acid into the vitreous chamber after NMDA treatment. We next misex-pressed basic helix loop-helix (bHLH) and homeobox genes in NMDA-treated retinas using a retroviral expression system. RESULTS: Muller glia of adult mammalian retinas proliferated in response to acute damage two days after NMDA treatment. These cells acquired a progenitor-like phenotype, and some of them migrated to the outer nuclear layer (ONL). A few of these cells expressed bipolar specific or rod photoreceptor specific markers. Retinoic acid treatment increased bipolar cell genesis. Misexpression of Math3 or NeuroD along with Pax6 promoted differentiation to amacrine cells. Co-expression of Crx and NeuroD promoted rod genesis. CONCLUSIONS: These findings demonstrated that retinal neurons regenerated even in adult mammalian retinas after toxic injury. We could partially control the fate of the regenerated neurons with extrinsic factors or intrinsic genes. PMID- 17134035 TI - [Physiological mechanism for the regulation of ocular circulation]. AB - Ocular circulation is regulated to maintain the homeostasis of retinal function in response to physiological stimulus. It is important to understand the mechanism of the regulation of ocular circulation under physiological conditions because the impairment of ocular circulation should cause severe retinal disorders. We previously investigated the physiological mechanism of ocular circulation in response to hypoxia, hypercapnia, and acute increased systemic blood pressure. In addition, it was reported that transpupillary thermotherapy might change ocular blood flow in patients with age-related macular degeneration. However, it still remains unclear whether the increased ocular tissue temperature may be associated with alteration of the ocular blood flow. Therefore, we examined the effect of ocular warming on retinal blood flow (RBF) and subfoveal choroidal blood flow (CBF) in humans. Ocular warming was induced in 10 healthy volunteers using an ocular warming lamp for 10 minutes. The RBF in the retinal artery and vein and the CBF in the foveal region were examined with a retinal laser Doppler velocimetry system and a laser Doppler flowmeter, respectively. Ocular warming increased RBF in the retinal artery and vein but decreased CBF in the foveal region. The CBF in the foveal region may contribute to maintaining a constant retinal temperature in response to ocular warming. PMID- 17134036 TI - [Cultured human corneal endothelial cell transplantation]. AB - Researchers have demonstrated the feasibility of transplanting human cultured corneal endothelial cells (HCEC) in various animal models. This review provides an overview of recent advances in our understanding of cultured corneal endothelial cell transplantation. We propose HCEC transplantation with a collagen sheet as the substitute carrier of HCEC. We also propose a novel strategy for corneal endothelial cell deficiency with the injection of adult human corneal endothelial precursors (HCEP). Using white rabbits or nude rats as keratopathy models, cultured HCEC were seeded on a collagen sheet. Descemetorhexis was performed on rabbit eyes. The HCEC collagen sheet was brought into the anterior chamber and fixed to the posterior stroma (HCEC group). Rabbit corneas with collagen sheet transplantation after descemetorhexis(collagen group) and with only descemetorhexis(no transplantation group) were the controls, respectively. As for HCEP transplantation, HCEP, isolated from rabbit corneal endothelial cells by sphere-forming assay, were injected into the anterior chamber and a face-down position was maintained for 24 hours in the rabbits (HCEP group). Pump function parameters of the HCEC sheets were 76-95% of those of human donor corneas. Mean corneal thickness in the HCEC group was significantly less than in the collagen and no transplantation groups 1, 3, 7, 14, 21, and 28 days (p< 0.05) after surgery. Cells were spread over the rear corneal surface in the HCEC group. In HE staining, marked stromal edema was present in the collagen and in the no transplantation groups, but not in the HCEC group with collagen sheets bearing monolayer cells. In the HCEP group, injected spheres were spread over the rear surface of the cornea and corneal edema was markedly suppressed. Our findings indicate that transplantation of cultured HCEC from adult human donor cornea by means of a collagen sheet can maintain the function of corneal dehydration. This suggests the feasibility of transplantation using cultured HCEC with a collagen sheet for corneal endothelial cell dysfunction. Additionally, adult precursor injection therapy can be also an effective strategy for corneal endothelial cell deficiency in place of conventional full-thickness corneal transplantation. PMID- 17134037 TI - [What can we learn from molecular genetic analyses of inherited eye diseases?]. AB - Over the past decade, the pace of gene identification of the causes of inherited eye diseases has increased dramatically as the complete human genome information becoming available. Molecular genetic analysis establishes reliable clinical diagnostic criteria and improves the accuracy of diagnosis. We encountered two atypical cases, a patient who had only bilateral band-shaped opacities without any gelatinous prominences, and a patient who had diffuse central corneal stromal opacity without any lattice lines. The diagnosis of gelatinous drop-like dystrophy and lattice corneal dystrophy I was confirmed by molecular genetic analysis of TACSTD2 and TGFBI, respectively. We confirmed the existence of a predominantly ocular type of stickler syndrome by identifying the mutation involving exon 2 of the COL2A1 gene in a Japanese patient who had received a diagnosis of rhegmatogenous retinal detachment. We surmise that in Japan in the past, this diagnosis may have been overlooked or misdiagnosed as Wagner disease. Molecular genetic analysis is also useful for gaining a better understanding of diseases. We detected a novel FZD4 mutation in a patient with familial exudative vitreoretinopathy who exhibited peripheral avascular areas bilaterally, a dragged disk, and retinal holes unilaterally, suggesting that FZD4 may be involved in the angiogenesis of the human peripheral retina. Molecular genetic analysis of a Japanese patient with pseudoxanthoma elasticum with choroidal neovascularization revealed a homozygous nonsense mutation in the ABCC6, a member of the ABC transporter family, indicating that angioid streaks may be caused by a primary metabolic disorder. The development of rapid and comprehensive genotyping systems using state-of-the-art technology such as genotyping microarray may eventually offer unique and reliable diagnostic tools. This should then accelerate our understanding of the basic mechanisms underlying inherited eye diseases and their phenotypic variability, thus facilitating prospective diagnosis. PMID- 17134038 TI - [Reading performance in patients with central visual field disturbance due to glaucoma]. AB - PURPOSE: To evaluate the relationship between central visual field disturbance due to glaucoma and reading performance. SUBJECTS AND METHODS: We evaluated the reading performance of 11 patients (14 eyes) with visual acuities of 1.0 and higher who had absolute scotoma within 3 degrees of the central visual field. The ages of the patients ranged from 37 to 77, with a median of 62 years. Maximum reading speed of texts printed horizontally and vertically was measured using MNREAD-J. Subjective reading difficulties were investigated orally. The relationship between the maximum reading speed of both horizontal and vertical texts, the subjective reading difficulties, the number of quadrants, and the position and continuity of absolute scotoma were evaluated. RESULTS: The mean maximum reading speed for vertical and horizontal texts by the 11 patients was 323 and 335 characters/minute(c/m), respectively, and was not significantly slower than that of normal eyes. In 2 of 14 eyes, a significant (p < 0.05) difference between maximum vertical and horizontal reading speeds was observed. In 3 of 14 eyes, absolute scotoma was distributed continuously in more than 2 adjacent quadrants, and these patients also had subjective reading difficulties. CONCLUSION: Reading difficulty is present in patients having absolute scotoma within 3 degrees of the central visual field when the scotoma involves more than 2 adjacent quadrants. PMID- 17134039 TI - [Three cases of corneal disorders associated with an oral anticancer drug S-1]. AB - PURPOSE: S-1 is a new oral anticancer drug containing tegafur, which is a prodrug of 5-fluorouracil. In this report, we describe the clinical features of three patients who suffered corneal disorders that seemed to be caused by S-1 administration. CASES: One female and two male patients, whose ages ranged from 57 to 69 years, were entered in this study. Between 3 to 14 months after they started oral S-1 therapy, they experienced sudden visual reduction. They all had corneal disorders, which occurred in the inferior and the superior areas, and progressed toward the center. As the corneal disorders invaded the pupil area, the patients noticed visual disturbance. They recovered their vision and their corneal disorders diminished when the drug was discontinued. However, after the drug intake was resumed, the corneal disorders occurred again in some cases. All patients also had lacrimal obstructions. CONCLUSION: S-1 treated patients should be followed carefully for ophthalmic complications because corneal disorders are likely to appear. PMID- 17134040 TI - [Opportunity, environment, and team work as contributions to ophthalmologic research]. PMID- 17134041 TI - [Change in terminology of macular degeneration and cataract]. PMID- 17134042 TI - Radiologic case study. Diagnosis: atlantoaxial instability secondary to rheumatoid arthritis. AB - Multiple elements contribute to the stability of the anterior C1-C2 articulation, making this region subject to pathologies including trauma, inflammation, infection, and congenital deformities. C1-C2 instability places a patient at risk for significant neurologic compromise. Radiologic imaging plays a fundamental role in diagnosing atlantoaxial instability, indicating etiology, showing details of associated abnormalities, and providing information for planning treatment. PMID- 17134043 TI - Professionalism: can we correct the problem from within? PMID- 17134044 TI - Manipulation of the shoulder using Codman's paradox. PMID- 17134045 TI - The use of minicondylar blade plates in the treatment of radial head and neck fractures. PMID- 17134046 TI - Focus on total ankle arthroplasty. PMID- 17134047 TI - New concepts in pertrochanteric hip fracture treatment. PMID- 17134048 TI - Pharmacokinetic alterations in obesity. AB - Although some medications have established dosing adjustments for obesity (Table 2), it remains unknown for the majority of medications if dosing adjustment is warranted. It is important to remember dosage adjustments may not be as simple as doubling an antibiotic dose because a patient is morbidly obese. Individualizing drug dosing is imperative in the obese, postoperative patient to ensure they simultaneously have therapeutic serum concentrations without drug toxicity. Much of what has been learned from studies in obese patients is that the pharmacokinetic alterations of medications are variable. Broad application of dosing guidelines even among medications within the same therapeutic class is likely not appropriate. An increased emphasis in researching the effects of obesity on the fate of medications is of paramount importance as the obese population grows. Practitioners should use caution and be vigilant in monitoring pharmacotherapy in obese individuals. PMID- 17134049 TI - Use of a distal radius plate for treatment of a tibial plafond fracture. PMID- 17134050 TI - The impact of irradiation on the microbiological safety, biomechanical properties, and clinical performance of musculoskeletal allografts. PMID- 17134051 TI - A practical review of the mechanisms of pain and pain management following ACL reconstruction. AB - The mechanisms of pain and pain management can be complex. Often while reading current research papers on the physiology of pain, it is easy to get lost in the minutia of the mechanisms being investigated. It is important that orthopedic surgeons have a general understanding of the mechanisms of pain, maintain a current understanding of the new areas of pain management, and continue to refine their treatment regimes. PMID- 17134052 TI - Outcomes following open repair of Bankart lesions for recurrent, traumatic anterior glenohumeral dislocations. AB - Though open stabilization using the Bankart procedure is still considered the "gold standard," few studies have examined the long-term outcomes. Between 1988 and 2001, 64 patients met inclusion criteria and had an open stabilization performed. Forty-one patients were available for follow-up at an average of 6.5 years. Forty (98%) patients reported having a stable shoulder. Average loss of external rotation was 4 degrees and average Rowe score was 96. This study further supports the trustworthiness of an open anatomic subcapsulolabral reconstruction for recurrent anterior instability and provides another standard from which to compare newer surgical methods. PMID- 17134053 TI - Short- and mid-term results after combined high tibial osteotomy. AB - In a consecutive series, 103 knees were treated with combined osteotomy. From these 103 knees, 80 knees were studied. Mean follow-up was 54.15 months (range: 13-96 months). After the first osteotomy is made 2 cm distally to the joint line, a bone wedge is removed based laterally. Its tip ends at the center of the tibial head/half bone wedge. The distal part of the tibia is placed into the valgus position and the half bone wedge is placed into the gap opened medially. The result was excellent in 44%, good in 45%, and poor in 11% of the knees. PMID- 17134054 TI - Rotator cuff interval reconstruction. AB - Five pairs of cadaveric shoulders underwent posterior and anterior drawer and inferior sulcus tests in five progressive conditions: intact, vented, following opening of the rotator cuff interval, reconstruction of the interval, and transfer of the coracoacromial ligament. The surgical treatments--vented, open rotator cuff interval, reconstruction, and coracoacromial ligament transfer--had an effect compared to the intact shoulders on the inferior stiffness (P = .00002) and on the anteroposterior stiffness (P = .00031). The difference between the stiffness of the reconstructed rotator cuff interval compared to the coracoacromial ligament transfer was significant for loading in the AP direction (P = .006) and for loading in the inferior direction (P = .005). PMID- 17134055 TI - Perioperative autotransfusion systems in TKA are not necessary. AB - Perioperative autotransfusion is believed to reduce postoperative allogenic transfusion. A retrospective review of 128 patients undergoing total knee arthroplasty with preoperative hemoglobin < 14 g/dL was performed. Group 1 received an intraoperative tourniquet and postoperative autotransfusion. Group 2 received a tourniquet and hemovac drain postoperatively. Group 3 had a tourniquet only during cementing with no drain. Nine (17%) patients in group 1, 9 (19%) patients in group 2, and 4 (15%) patients in group 3 required transfusion. Perioperative autotransfusion does not decrease the need for allogenic transfusion when compared to similar patients at risk for transfusion because of preoperative hemoglobin level < 14 g/dL. PMID- 17134056 TI - Pyomyositis of the shoulder girdle. PMID- 17134057 TI - Tuberculous osteomyelitis of the hand in a child. PMID- 17134058 TI - Rapid progression of metastatic bronchogenic carcinoma after felon drainage. PMID- 17134059 TI - Prevention of sudden cardiac death in the young: targeted evaluation of those at risk. PMID- 17134060 TI - QT dispersion: comparison between diabetic and non-diabetic individuals and correlation with cardiac autonomic neuropathy. AB - INTRODUCTION: The QT interval on the resting electrocardiogram (ECG) expresses the myocardial depolarisation and repolarisation time. Elevated values of QT dispersion (QTd) are associated with cardiovascular mortality in diabetics. Cardiac autonomic neuropathy (CAN) is a common complication of diabetes that is also associated with increased morbidity and mortality. However, there are no data in the literature concerning the relation between CAN and QTd in diabetics. The aim of this study was to investigate: 1) the differences in QTd between diabetics and non-diabetics; 2) the differences in QTd between those with type 1 and type 2 diabetes; 3) the relation between QTd and CAN. METHODS: The study population included 184 diabetics (63 type 1, group D1; 121 type 2, group D2) and 100 healthy controls who had similar age and sex distribution to D1 (n=44) and D2 (n=56) subjects. CAN assessment was made using the standard Ewing and Clarke tests. The QT interval was measured on the 12-lead resting ECG. QTd was calculated automatically using special software. RESULTS: QTd values did not differ significantly between controls and D1 (p=0.15) or D2 (p=0.27). QTd was significantly greater in D2 than in D1 (p=0.02). There was no significant difference in QTd between those with and without CAN in either group of diabetics. CONCLUSIONS: QTd values do not differ between individuals with and without diabetes. Type 2 diabetes is associated with higher QTd values than is type 1 diabetes. CAN does not affect QTd in diabetics. PMID- 17134061 TI - The acute and chronic effect of cigarette smoking on the elastic properties of the ascending aorta in healthy male subjects. AB - INTRODUCTION: Aortic elastic properties are markers and prognosticators of cardiovascular risk. Smoking adversely affects arterial elastic properties. The acute and chronic effect of cigarette smoking on the elastic properties of the ascending aorta has not been determined. METHODS: The chronic part of the study included 130 healthy male subjects (age 38.1+/-8.9 years, 85 smokers and 45 non smokers) with no other risk factors for cardiovascular disease. Twenty of the smokers (age 40.0+/-4.2 years) were also included in the acute study. In the acute study each subject was studied on two occasions: one while smoking one cigarette and the other during sham smoking. Ascending aortic distensibility was assessed by two-dimensional, guided, M-mode transthoracic echocardiography of the aortic root at baseline and at the end of smoking. In the chronic study the association was assessed between smoking status and ascending aortic distensibility. RESULTS: Smoking acutely increased systolic, diastolic, and pulse pressure. Furthermore, smoking decreased ascending aortic distensibility (by 0.53+/-0.86 cm(2) x dyne(-1) x 10(-6), p<0.05). The chronic study showed that smokers had greater ascending aortic distensibility than non-smokers (2.22+/-0.93 vs. 2.75+/-1.07 cm(2) x dyne(-1) x 10(-6), respectively, p<0.05). Aortic distensibility was independently associated with smoking (beta=-0.293, p<0.001) after controlling for possible confounders. No association of aortic distensibility with the intensity or duration of smoking was observed within the group of smokers. CONCLUSIONS: Cigarette smoking results acutely in a deterioration of ascending aortic elastic properties in healthy male subjects. On a chronic basis, smokers have decreased ascending aortic distensibility compared to non-smokers. This effect of smoking on aortic distensibility is not related to dose or duration. PMID- 17134062 TI - Clinical and prognostic value of elevated CA125 levels in patients with congestive heart failure. AB - BACKGROUND: Increased serum levels of carbohydrate antigen 125 (CA125), a tumor marker associated with ovarian cancer, have also been reported in other malignant and non-malignant diseases. We assessed the correlation of the CA125 serum levels with the severity of congestive heart failure (CHF) and investigated their potential prognostic value in relation to major cardiovascular events. METHODS: CA125 levels were measured in 95 male patients aged 70+/-10 years, admitted for decompensated CHF. The patients were divided into three groups, according to their New York Heart Association (NYHA) functional class. Group A contained 23 patients in NYHA IV, group B 34 patients in NYHA III, and group C 38 patients in NYHA I-II. The patients were also divided into two groups according to their CA125 value on admission. Group 1 included 45 patients with normal CA125 levels and group 2 50 patients with elevated CA125. All patients were followed for 15+/ 8.5 months and the major cardiovascular events (death and re-hospitalizations due to CHF) were recorded. RESULTS: Serum levels of CA125 were higher in groups A and B than in group C (36.4 [19.8-82] U/ml and 34.6 [26-78] U/ml vs. 25.3 [9.1-29] U/ml, respectively, p<0.05). No correlation was detected between CA125 levels and left ventricular ejection fraction. However, patients with pulmonary congestion and peripheral edemas had higher levels of CA125 (p=0.002 and p<0.03, respectively). Nineteen patients died during the follow-up period, but the mortality rate was not significantly different between groups 1 and 2 (p=0.8). Nevertheless, the patients of group 1 reported fewer re-hospitalizations than patients of group 2 (p=0.003). The relative risk (RR) for re-hospitalization was calculated to be RR: 0.4, 95% CI: 0.215-0.76 (p<0.005), in patients with elevated levels of CA125. Cox regression analysis revealed that CA125 had independent prognostic value (OR: 1.007 [95% CI: 1.004-1.010], p<0.0001) for re hospitalizations. CONCLUSION: Serum levels of CA125 are associated with the severity of CHF and are also independent predictive markers for re hospitalizations. We therefore conclude that CA125 can be used as a prognostic marker of disease severity and increased morbidity in patients with decompensated CHF. PMID- 17134063 TI - Heart disease and pregnancy. PMID- 17134064 TI - Cost-effectiveness of drug-eluting stents. PMID- 17134065 TI - Should atenolol still be recommended as first-line therapy for primary hypertension? PMID- 17134066 TI - The natural history of a penetrating atherosclerotic ulcer at the aortic arch. PMID- 17134067 TI - Subacute thrombosis following implantation of zotarolimus-eluting stent. AB - Endeavor (Medtronic Europe SA) is a new zotarolimus-eluting stent with a favourable safety profile reported in early trials. We report a case of subacute thrombosis following deployment of an Endeavor stent in the distal left anterior descending coronary artery, despite optimum loading with and continuation of clopidogrel therapy as well as previous administration of a IIb/IlIa inhibitor. PMID- 17134068 TI - Lyme carditis: complete atrioventricular dissociation with need for temporary pacing. AB - Lyme borreliosis is a tick-borne disease. Cardiac manifestations of the disease are extremely rare. We report a case of Lyme carditis in an otherwise healthy male, who presented to the Accident & Emergency Department with chest pain, dizziness and generally symptoms indicating ischaemic heart disease. This patient, without documented history of Lyme disease, acutely developed third degree atrioventricular block, which required placement of a transvenous pacemaker and resolved when the patient was administered doxycycline. PMID- 17134069 TI - The role of mitral valve repair in heart failure. AB - We report a 71-year-old man with severe mitral valve regurgitation in end stage heart failure intractable to medical treatment. He underwent mitral valve repair, coronary artery bypass grafting and tricuspid annuloplasty, and his postoperative course was uneventful with significant improvement of his symptoms. Mitral valve repair should be considered as a treatment modality in patients with cardiac failure and significant mitral valve regurgitation. PMID- 17134070 TI - Therapy in cardiovascular disease: it is time for therapeutic classification. PMID- 17134071 TI - Women in cardiovascular medicine. PMID- 17134072 TI - Drugs that cause photosensitivity. PMID- 17134073 TI - The try-in visit for restorative dentistry. AB - The try-in visit for restorative dentistry is one of many important steps toward excellence. Attention to detail in all parts of this visit outlined above will lead to fewer remakes and more satisfied patients and dentists. PMID- 17134074 TI - An oral complication in a patient with cyclic vomiting syndrome: a case report and review. AB - Cyclic vomiting syndrome (CVS) is a disorder seen primarily in children. It is characterized by a number of clinical symptoms; the most prominent are recurrent, self-limited episodes of intractable nausea and vomiting that are not caused by any specific disease. CVS can be severely debilitating and may interfere with an individual's quality of life. This article describes a patient with CVS who sought treatment for complications after dental extractions, reviews important aspects of this disease, and discusses its potential significance as it relates to the present case. PMID- 17134075 TI - Technique for placement of a posterior prefabricated fiber-reinforced composite bridge. AB - Fiber-reinforced composite (FRC) materials currently are used as alternatives for fixed restorations of edentulous areas within the posterior and anterior regions of the mouth. A chairside technique, using a prefabricated FRC framework, allows the clinician to offer the patient another fixed option for replacing a missing posterior tooth that is more time-efficient and cost-effective than other, more traditional approaches. Procedures that incorporate adhesive dentistry give dentists the ability to replace missing teeth and splint unstable teeth for periodontal or orthodontic purposes. This article demonstrates the sequence and chairside technique for placing a prefabricated FRC framework to restore a posterior edentulous area. PMID- 17134076 TI - Success of root canal therapy in HIV-positive patients. AB - This retrospective study evaluated the success rate of root canal therapy in 157 HIV-positive patients who had undergone nonsurgical endodontic treatment in a dedicated clinic between 1998 and 2004. Dental records and radiographs were reviewed and information concerning age, gender, medical history, anti-retroviral medication, treatment dates, and follow-up evaluation was compiled. All root canal treatment was performed following the clinical guidelines of the American Association of Endodontists. A single practitioner who was responsible for seeing all of the patients at the six-month recall examination reviewed radiographs. Success was defined as the absence of pain and swelling as well as the absence of the periapical lesion (or a reduction in size compared to its preoperative radiographic size). At the six-month postoperative evaluation, a success rate of 90% was observed in the study group. No statistically significant differences were noted when the success of the root canal therapy was related to the symptomatic clinical presentation, the antiretroviral therapy, or the viral load. PMID- 17134077 TI - The effect of a mouthrinse containing essential oils on dental restorative materials. AB - Mouthrinses that contain essential oils are effective for controlling plaque and periodontal disease. Recent studies have shown that such mouthrinses are effective at preventing the formation of biofilm in dental unit waterlines. However, there is no information in the literature regarding the effect of such mouthrinses on restorative materials used within the oral cavity. Specimens of three common restorative materials (a glass ionomer, a composite resin, and amalgam) were subjected to continuous exposure to Listerine and distilled water for 10 days; at that time, the strength, fluid sorption, and surface appearance of the specimens were compared. Specimens of the test materials also were placed in intraoral devices; volunteer patients wore these devices for 12 hours per day for a period of 10 days. During that time, the patients were instructed to rinse twice daily for 30 seconds with Listerine Cool Mint or a non-active mouthrinse. After 10 days, the specimens were salvaged from the devices and inspected by visible and SEM examination. This study indicates that routine use of mouthrinses containing essential oils (or even prolonged exposure to such mouthrinses) has no adverse effects on restorative materials that might be expected to react to such mixtures because of their chemical compositions. It was concluded that active mouthrinses do not appear to have any adverse effects on a variety of restorative biomaterials. PMID- 17134078 TI - Shear bond strength of indirect resin to NiCr with different metal surface preparations. AB - This study sought to evaluate the shear bond strength of indirect resin to nickel chromium (NiCr) after different surface preparations. Sixty-four metal samples were divided into four groups, according to surface preparation, and two layers of opaque resin and three layers of body resin were applied. Each group was divided into two subgroups based on storage period in distilled water at 37 degrees C. Mechanical shear testing was performed using a universal testing machine with 0.5 mm/minute of cross-head speed. Debonded areas were analyzed and classified according to different failure types, such as cohesive, adhesive, and mixed. The storage periods were compared using a student T-test (alpha = 0.05); Group T (that is, the tin electroplating group) showed the highest average after 24 hours. The macro retentions showed higher shear bond strength than the sandblasted samples (p < 0.05). Tin electroplating did not improve the shear bond strength of metal-resin bonding. The results were not affected by 30-day storage, with the exception of group T (p < 0.05). PMID- 17134079 TI - Root canal morphology of the mesiobuccal root in maxillary first molars of a Jordanian population. AB - This in vitro study investigated the presence of second (MB2) root canals and the number of apical foramina found in mesiobuccal roots of the permanent maxillary first molars; in addition, the study evaluated the effectiveness of magnification to detect MB2 canals. One hundred extracted maxillary first molars were collected from Jordanian patients. The number of roots (as well as their morphology) was investigated. The number of canal orifices in mesiobuccal root was assessed, with and without magnification, and findings were compared. The number of apical foramina and the distance between mesiobuccal and second canal orifices were investigated under magnification. Of the 97 specimens that were subject to additional investigation, all teeth had three separated roots, except for three specimens that had either mesiobuccal or distobuccal roots fused with the palatal root. The number of MB2 canals that were detected increased from 55 (56.7%) to 61 (62.9%) teeth when magnification was used. Within the limits of this in vitro study, it was concluded that the Jordanian population had a high percentage of MB2 canals in the mesial buccal roots. Moreover, the use of clinically used magnifying devices increased the number of MB2 canals detected. PMID- 17134080 TI - Multiple hamartoma syndrome (Cowden's syndrome): case report and literature review. AB - Multiple hamartoma syndrome (also known as Cowden's syndrome) is a genetic disorder. The signs and symptoms of this disorder often do not appear concurrently or with the same degree of severity. While much of the literature regarding Cowden's syndrome (CS) focuses on the dermatologic and gastro enterologic features of the disease, there also are unique oral and facial symptoms that manifest early in this condition. This disorder might be overlooked and underdiagnosed by the dental community. There is a strong link between CS and female breast cancer, thyroid cancer, gastrological polyps, and rare forms of brain cancer; as a result, it is essential for the dental community to gain a greater understanding of this syndrome and its unique oral manifestations. PMID- 17134081 TI - Endodontic treatment of a three-rooted maxillary first premolar: a case report. AB - Identifying and accessing all pulp canals is particularly challenging in the endodontic treatment of teeth with atypical canal configurations. This report describes the nonsurgical endodontic treatment of a maxillary first premolar with three root canals. The prevalence of this unusual morphology is reviewed and suggestions are offered to help dentists identify and treat such teeth. This case reminds dentists to be alert for variations in the root canal anatomy of maxillary first premolars. PMID- 17134082 TI - Unilateral tongue atrophy. AB - A 64-year-old woman who sought routine dental treatment was noted to have unilateral atrophy of the tongue. The medical and dental significance of this manifestation is reviewed. PMID- 17134083 TI - Six mandibular permanent incisors: report of a case. AB - This article involves dental management of a girl with bilateral supplemental mandibular permanent incisors. If not removed promptly, these supernumerary teeth will lead to crowding and malocclusion; as a result, early diagnosis is important. Dentists who treat children should be aware of this condition when unusual crowding and displacement is seen in the mandibular incisor region. PMID- 17134084 TI - The methamphetamine epidemic and dentistry. AB - Methamphetamine is a potent central nervous system stimulant with limited therapeutic effects. This drug produces prolonged euphoria and is relatively inexpensive to purchase and easy to make and distribute. Methamphetamine changes normal physiologic processing of several centrally acting neurotransmitters and ultimately leads to neurotoxicity and neurodegeneration from chronic use. Chronic methamphetamine use has been associated with severe oral health effects; rampant caries is the most notable of these. Dental professionals must recognize patients who are involved with methamphetamine use and understand the risk factors associated with its deleterious oral effects so that preventive and treatment strategies may be implemented for patients who use this drug. PMID- 17134085 TI - Removable white lesions of buccal mucosa. Candidiasis. PMID- 17134086 TI - Medical regulation: promoting excellence and safety. PMID- 17134087 TI - Vaccines against human papillomavirus: the future is bright. PMID- 17134088 TI - Facial flaps for facial skin lesions. PMID- 17134089 TI - Pinnaplasty: the correction of the prominent, protruding or lop ear. PMID- 17134090 TI - Techniques of nasal reconstruction. PMID- 17134091 TI - Stroke: improving outcome through better diagnosis and treatment. PMID- 17134092 TI - Human rabies and its prevention. PMID- 17134093 TI - Principles of paediatric injuries. PMID- 17134094 TI - The carpal tunnel. PMID- 17134095 TI - Why good communication is worth the effort. PMID- 17134096 TI - Phlebotomy tips for junior doctors. PMID- 17134097 TI - Deliberate self harm and assessing suicidal risk. PMID- 17134098 TI - Should junior doctors be obtaining consent? PMID- 17134099 TI - So you want to be...a neurologist. PMID- 17134100 TI - Who needs an implantable cardioverter defibrillator? PMID- 17134101 TI - A qualitative evaluation of the trainees' experience of assessments in the MMC foundation programme. PMID- 17134102 TI - The ligated ureter: beware or be aware. PMID- 17134103 TI - 'Not just an ordinary bump to the head': traumatic superficial temporal artery false aneurysm. PMID- 17134104 TI - Myositis ossificans progressiva. PMID- 17134105 TI - Right atrial myxomas and hairy cell leukaemia: coincidence or causal relationship? PMID- 17134107 TI - The first anaesthetic. PMID- 17134108 TI - Capacity to consent: clinical complexities. PMID- 17134109 TI - Pension planning for your family: redistribution of wealth. PMID- 17134110 TI - Should non-steroidal anti-inflammatory drugs be given to neurosurgical patients? PMID- 17134111 TI - Molecular pathogenesis of primary central nervous system lymphoma. AB - Primary central nervous system lymphoma (PCNSL) is an aggressive form of non Hodgkin lymphoma (NHL) typically associated with a worse prognosis than other localized extranodal lymphomas with similar histological characteristics. The defining feature of PCNSL is its confinement to the central nervous system (CNS), with proclivity for growth within the leptomeningeal as well as intraocular compartments. Primary CNS lymphoma rarely disseminates outside the CNS and accounts for less than 5% of all primary brain neoplasms. At least 95% of PCNSLs are of large B-cell histology, the most common subtype of NHL. Consistent with the trend seen in systemic NHLs, the incidence of PCNSL has markedly increased over the past three decades, both in immunocompromised and immunocompetent patients. Because PCNSL is relatively rare, the identification of molecular prognostic biomarkers and the definition of a standard therapeutic strategy have been challenging. The authors discuss the current knowledge of the molecular pathogenesis of CNS lymphomas and review the recent advances in gene expression profile analysis and identification of novel prognostic biomarkers. PMID- 17134112 TI - Primary diffuse large B-cell lymphoma of the dura mater and cranial vault. Case report and literature review. AB - Primary high-grade lymphoma of the dura mater and cranial vault has rarely been reported. The authors treated a 61-year-old man who presented with a slow-growing scalp mass that involved the cranial vertex. Magnetic resonance imaging revealed an oval mass of the dural type with peripheral edema in the bilateral parietal region, with attachment to the cranial vault and extension to the subgaleal space. After subtotal resection, pathological examination yielded a diagnosis of malignant large B-cell lymphoma. Twenty-three months postoperatively, after undergoing radiation therapy and chemotherapy, the patient is neurologically intact and without systemic dissemination of the malignancy. This is a case of primary malignant B-cell lymphoma of the dura mater with extensive involvement of the skull, which is a very rare event. Imaging-based diagnosis and combined therapy consisting of surgery, radiation therapy, and chemotherapy for the disease are discussed, and the literature on extraaxial malignant lymphomas is extensively reviewed. PMID- 17134113 TI - Implications of the blood-brain barrier in primary central nervous system lymphoma. AB - The optimal treatment of primary central nervous system lymphoma (PCNSL), a rare form of extranodal non-Hodgkin lymphoma, has yet to be defined. Whole-brain radiation therapy (WBRT) has limited efficacy as a single therapeutic modality and is associated with a high risk of delayed neurotoxicity. Methotrexate-based chemotherapy regimens yield poor drug penetration across the blood-brain barrier (BBB), thus necessitating administration of high doses with the concomitant risk of increased systemic and neurological toxicity. Combined-modality therapy (WBRT plus chemotherapy) can improve response and survival rates, yet it is associated with a high risk of neurotoxicity. The aim of chemotherapy in conjunction with BBB disruption is to maximize drug delivery to the brain and improve the agent's efficacy, while preserving neurocognitive function and minimizing systemic toxicity. Methotrexate-based chemotherapy regimens administered in conjunction with BBB disruption have shown promising results in PCNSL. Animal models of central nervous system lymphoma and drug neurotoxicity offer new possibilities to study the effects of various treatments on PCNSL and normal brain and can also help understand biological and pathophysiological aspects of this disease. Because the intact BBB is even less permeable to antibodies than it is to drugs, preclinical and clinical studies of monoclonal antibody delivery (for example, rituximab and 90Y ibritumomab tiuxetan) to the brain in conjunction with BBB disruption offer a new possibility to make these novel treatments more efficient against PCNSL. Regarding the evaluation of more sensitive and specific diagnostic imaging tools, iron oxide-based contrast agents for magnetic resonance imaging have shown promise for better differentiation of PCNSL from other white matter diseases. PMID- 17134114 TI - Chemotherapy for primary central nervous system lymphoma. AB - Chemotherapy, with or without radiotherapy, is the mainstay of treatment for primary central nervous system lymphoma (PCNSL). High-dose methotrexate (MTX) is the most effective drug available to treat these lesions, and it is used in doses of 1 to 8 g/m(2), either as a single agent or in combination with other drugs such as corticosteroid agents, cytarabine, procarbazine, vincristine, carmustine, lomustine, thiotepa, cyclophosphamide, temozolomide, and rituximab. To date, an overwhelming number of different regimens in which high-dose MTX is used have been reported. Given the lack of randomized trials, however, the optimal treatment remains controversial. Varying methodology makes the comparison of available studies extremely difficult, yet some common themes can be found throughout the literature. Treatment paradigms vary considerably according to the patient's age. Most studies support the use of chemotherapy-only treatments for elderly patients (> 60 years), given the high risks of neurotoxicity associated with radiotherapy. Nevertheless, the prognosis remains poor regardless of the chemotherapy chosen, and less toxic regimens might be preferable for such elderly patients. Conversely, in younger patients (< 60 years), there is growing evidence that commonly used chemotherapy-only regimens are associated with increased relapse rates that may not justify deferral of radiotherapy. Thus, a significant focus of research has been the development of intensified chemotherapy regimens that could replace radiotherapy. In this article, the authors discuss the principles guiding the use of chemotherapy for PCNSL, and critically review the available literature, including the most recent trials. PMID- 17134115 TI - Surgical interventions for primary central nervous system lymphoma. AB - Early diagnosis is central to proper management of primary central nervous system lymphomas (PCNSLs). Surgical intervention hinges on initial entertainment of a diagnosis of a PCNSL, based on acute neurological presentation and neuroimaging findings. Unless there is an urgent need for surgical decompression, a biopsy to obtain a diagnosis of PCNSL is the first step in surgical management. Repeated biopsy may be necessary in patients who have received preoperative steroid therapy. Patients with PCNSL may also present with leptomeningeal involvement, resulting in the need for an Ommaya reservoir for intrathecal chemotherapy. In cases in which hydrocephalus develops, placement of a ventriculoperitoneal shunt may be necessary. Two case studies are presented to highlight the role of surgical intervention in PCNSL. PMID- 17134116 TI - Efficacy of celecoxib in the treatment of CNS lymphomas: an in vivo model. AB - OBJECT: The incidence of primary central nervous system lymphomas (PCNSLs) has increased over the past several decades. Unfortunately, even with the most effective therapeutic regimen (that is, methotrexate with wholebrain radiation therapy), PCNSL recurs within a few years in more than half of the treated patients and is eventually fatal. Because PCNSL usually occurs in older patients and in those with acquired immunodeficiency syndrome, combination treatments in which both chemo- and radiation therapy are used is often poorly tolerated and results in a significant reduction in the quality of life. Recently, it has been demonstrated that the selective cyclooxygenase- 2 inhibitor celecoxib (Celebrex), can block the growth of lymphoma cells in vitro. METHODS: To create an experimental animal model in vivo for the PCNSL study, the authors intracranially injected a human B-cell lymphoma cell line into nude mice. Their data demonstrate that this experimental model is an excellent one for human PCNSL with brain and leptomeningeal involvement. They also evaluated the feasibility of using celecoxib as a therapeutic agent in the treatment of PCNSL. Nude mice with intracranial lymphomas were treated with celecoxib contained in the animal chow. The treated animals demonstrated significantly prolonged survival times compared with the untreated animals. CONCLUSIONS: Based on the authors' data, celecoxib may be a promising therapeutic agent for the treatment of PCNSL. PMID- 17134117 TI - Primary central nervous system lymphoma: presentation, diagnosis and staging. AB - Primary central nervous system lymphoma (PCNSL) is a rare form of non-Hodgkin lymphoma that affects the brain, spinal cord, leptomeninges, and eyes. The clinical presentation and neuroimaging appearance of PCNSL differ in immunocompetent patients and in those with acquired immunodeficiency syndrome (AIDS). A magnetic resonance (MR) image of the brain in immunocompetent patients with PCNSL typically demonstrates one or more homogeneously enhancing lesions located in the periventricular white matter, characteristically spanning the corpus callosum. In patients with AIDS, multiple ring-enhancing lesions are more common. After neuroimages raising the suspicion of PCNSL are obtained, a definitive diagnosis should be established in both immunocompetent and AIDS patients by performing pathological analysis of cerebrospinal fluid (CSF), vitreous fluid, or a biopsy specimen. Brain biopsy sampling remains the gold standard for PCNSL diagnosis in all patients, although the possibility of establishing routine, minimally invasive diagnostic procedures in which Epstein Barr virus polymerase chain reaction (PCR) analysis of the CSF and nuclear imaging are used is currently under investigation in the population of patients with AIDS. At the time of diagnosis, the patient should undergo further evaluation, which should include a physical examination, ophthalmic evaluation with a slit-lamp examination, serum lactate dehydrogenase levels, human immunodeficiency virus testing, computed tomography scans of the chest/abdomen/pelvis, bone marrow biopsy sampling, contrast-enhanced brain MR imaging, and lumbar puncture (LP). Testicular ultrasonography studies should be considered in men. In patients who cannot undergo LP or in those with evidence of spinal cord dysfunction, contrast-enhanced MR imaging of the entire spine should be considered. PMID- 17134118 TI - Pathology of primary central nervous system lymphoma. AB - The understanding of primary central nervous system lymphoma (PCNSL) has lagged behind that of the much more common systemic, nodal lymphomas. Reasons for this include the relative rarity of PCNSL and the fact that these lesions differ in some ways from their histologically similar systemic counterparts. The purpose of this paper is twofold: first, the author provides concise descriptions of the pathological features of both common and uncommon types of PCNSL while discussing issues such as the confusion surrounding histological classification of PCNSLs, problems related to intraoperative diagnosis, and the appropriate diagnostic work up. Second, the author discusses a small number of molecular studies to demonstrate the great promise they offer. Such studies have already clarified some issues, including the category of lymphocyte from which the majority of PCNSLs are derived. Hopefully in the future these studies will help guide treatment decisions. PMID- 17134119 TI - Distinguishing primary central nervous system lymphoma from other central nervous system diseases: a neurosurgical perspective on diagnostic dilemmas and approaches. AB - OBJECT: White matter diseases, including demyelinating or inflammatory disorders, may be indistinguishable clinically and radiologically from some central nervous system (CNS) tumors. In such situations, determination of the final diagnosis is difficult. An example is the differential diagnosis of non-acquired immunodeficiency syndrome-related primary central nervous system lymphoma (PCNSL) and multiple sclerosis (MS), a demyelinating disease. Unfortunately, delayed diagnosis and treatment of PCNSL can negatively affect prognosis. METHODS: The authors reviewed the cases of eight patients with PCNSL or MS. In each case, the initial diagnosis (PCNSL or MS) was equivocal. In these cases, conventional diagnostic approaches were not definitive, thus further delaying diagnosis. The initial symptoms, the selected diagnostic tests, and the presumptive as well as final diagnosis for each case are discussed. The final diagnosis was PCNSL in six cases and MS in two. The uncertainty about the clinical or initial pathological presentation required further diagnostic evaluation in all cases. Two important neurosurgical guidelines are the avoidance of corticosteroid agents and performance of biopsy sampling rather than volumetric tumor resection. Highvolume lumbar puncture, slit-lamp examination/vitrectomy, new CNS imaging techniques, and repeated biopsy procedures also proved helpful. CONCLUSIONS: In PCNSL, early definitive diagnosis and treatment are the keys to successful outcomes. Knowledge of strategies essential to early diagnosis lessens the need for brain biopsy sampling, but this procedure is still usually necessary. In such selected cases, biopsy sampling is appropriate even when pathological investigation shows MS rather than PCNSL. Complete resection is not indicated in PCNSL and can lead to additional sequelae. PMID- 17134120 TI - Imaging of primary central nervous system lymphoma. AB - Primary central nervous system lymphoma (PCNSL) is an aggressive neoplastic process that occurs in both immunocompetent and immunocompromised patients. Over the past 30 years there has been a steady increase in the number of cases in both patient populations. The imaging features for the disease and demographic characteristics within these patient populations vary, and in this article the authors describe the salient features of these two groups. PMID- 17134121 TI - Primary dural lymphomas: a review. AB - Primary dural lymphoma (PDL) is a rare subtype of primary central nervous system lymphoma (PCNSL) that arises from the dura mater and differs biologically from other CNS lymphomas. The PDL is usually a low-grade marginal zone lymphoma (MZL), whereas other types of PCNSLs are usually high-grade, diffuse, large B-cell lymphomas. A PDL usually presents with single or multiple extraaxial masses that enhance diffusely with addition of contrast material and can be confused with meningioma. The MZLs respond well to local therapy such as surgery and radiation treatment. Most patients attain complete response and have good local disease control. Nevertheless, the risk of systemic relapse appears to be high, probably because the dura mater is outside of the blood-brain barrier. PMID- 17134122 TI - Lymphomatous meningitis in primary central nervous system lymphoma. AB - Lymphomatous meningitis (LM) due to primary central nervous system (CNS) lymphoma is an uncommon problem in neurooncology and can occur at time of diagnosis or recurrence. Notwithstanding frequent focal signs and symptoms, LM is a disease affecting the entire neuraxis, and therefore staging and treatment need to encompass all cerebrospinal fluid (CSF) compartments. Central nervous system staging of LM includes contrast agent-enhanced cranial computed tomography (CT) or Gd-enhanced magnetic resonance (MR) imaging, Gd-enhanced spinal MR imaging, CT myelography, and radionuclide CSF flow study. Treatment of LM includes involved field radiotherapy of bulky or symptomatic disease sites and intra-CSF drug therapy. The inclusion of concomitant systemic therapy can benefit patients with LM and can obviate the need for intra-CSF chemotherapy. At present, intra-CSF drug therapy is confined to three chemotherapeutic agents (methotrexate, cytosine arabinoside, and thiotepa) administered by a variety of schedules either by intralumbar or intraventricular drug delivery. Although treatment of LM is palliative and the expected median survival of patients is 4 to 6 months, it often provides stabilization and protection from further neurological deterioration. In patients with primary CNS lymphoma, CNS prophylaxis has been recommended (using a combination of high-dose systemic chemotherapy and intra-CSF chemotherapy), but the strategy remains controversial because high-dose systemic methotrexate is commonly used as an adjuvant therapy. Patients with primary CNS lymphoma at high risk as defined by positive CSF cytology or neuroradiography consistent with LM may benefit from the inclusion of intra-CSF chemotherapy. PMID- 17134123 TI - Rare pathological variants and presentations of primary central nervous system lymphomas. AB - Primary central nervous system lymphoma (PCNSL) is a rare form of primary brain neoplasm, accounting for less than 3% of all primary brain tumors. Ninety percent of cases involve a large B-cell lymphoma that presents as a homogeneously enhancing lesion or lesions, typically deep-seated in the brain parenchyma. The authors describe unusual pathological forms of PCNSLs, including low-grade, T cell, and Burkitt types, and also rare presentations such as neurolymphomatosis and pituitary lymphomas. PMID- 17134124 TI - Primary central nervous system lymphoma in children. AB - Primary central nervous system lymphoma (PCNSL) is a very rare brain tumor in children, and the optimal management and prognosis of such patients have yet to be defined. In this study, the incidence rate, clinical features, diagnosis, and treatment of childhood PCNSL are reviewed. Except for human immunodeficiency virus-related PCNSL, the prognosis for patients with this tumor type is significantly better in children than in adults. In the absence of prospective studies, it is very difficult to determine the true incidence and the best therapeutic strategy for this rare entity. The majority of children with PCNSL, however, can achieve long-term remissions with intensive chemotherapy alone (an estimated 70% 5-year event-free survival rate), and cranial irradiation can be reserved for relapse of the disease. Further progress in the management of childhood PCNSL will require prospective multinational studies. PMID- 17134125 TI - Ocular manifestations and treatment of central nervous system lymphomas. AB - Intraocular primary central nervous system lymphoma (PCNSL), also called primary intraocular lymphoma (PIOL), is a subset of PCNSL in which lymphoma cells invade the subretinal pigment epithelial space and vitreous cavity with or without central nervous system involvement at the time of ocular diagnosis. The frequency of this rare condition has increased over the past years in immunosuppressed as well as immunocompetent patients. The authors review the current status of PIOL and elaborate on their group's experience with its diagnosis and treatment. The incidence of PIOL is increasing. There is evidence that chronic antigenic stimulation may result in the development of PIOL. Recent advancements in the diagnosis of PIOL include better handling of vitreous specimens for cytological studies, immunocytological investigation for lymphoid cells, flow cytometry, cytokine evaluation, and molecular analysis. Because PIOL has a nonspecific presentation, the differential diagnosis should include infectious and noninfectious causes presenting with vitreitis and/or subepithelial infiltration as well as paraneoplastic syndromes including CRMP-5 optic neuropathies. Given that therapy is long-term and has significant systemic and ocular complications, tissue diagnosis is important. Treatment of PIOL may include systemic chemotherapy in which high-dose methotrexate-based regimens are used as well as intraocular injections of methotrexate and rituximab (anti-CD20 antibody). Cranial and ocular external-beam radiotherapy is being used less often today. Further studies are needed to prevent the tumor formation in terms of eliminating antigenic load and inhibiting Bcell chemokines as well as to determine the optimal local and systemic chemotherapy and immunotherapy options in the management of PIOL. PMID- 17134126 TI - Controlling desensitized states in ligand-receptor interaction studies with cyclic scanning patch-clamp protocols. AB - Ligand-gated ion channels are important control elements in regulation of cellular activities, and increasing evidence demonstrates their role as therapeutic targets. The receptors display complex desensitization kinetics, occurring on vastly different time scales. This is not only important in biology and pharmacology but might also be of technological significance since populations of receptors under microfluidic control can function analogously to DRAM memory circuits. Using a novel microfluidic method, and computer modeling of the receptor state distributions, we here demonstrate that GABAA receptor populations can be controlled to display high or low EC50 values, depending on input function (i.e., the exact pattern of agonist application). The sensitivity of the receptors can be tuned up to 40-fold (beta-alanine) by the particular agonist exposure pattern. By combining patch-clamp experiments with computer modeling of receptor state distributions, we can control the assembly of receptors in desensitized states. The technique described can be used as an analytical tool to study the effect of desensitization on the activity of ion channel effectors. We describe the differential blocking effect of the competitive antagonist bicuculline on the high- and low-EC50 GABAA receptor preparations and conclude that the inhibition is dramatically dependent on how the different desensitized states are populated. Furthermore, we show that both GABA and beta-alanine, two agonists with different affinity but similar efficacy, induce the same type of desensitization behavior and memory effects in GABAA receptors. PMID- 17134127 TI - Metabolomics: The greatest omics of them all? PMID- 17134128 TI - Combining fluorescence detection and mass spectrometric analysis for comprehensive and quantitative analysis of redox-sensitive cysteines in native membrane proteins. AB - Monobromobimane (MBB) is a lipophilic reagent that selectively modifies free cysteine residues in proteins. Because of its lipophilic character, MBB is capable of labeling cysteine residues in membrane proteins under native conditions. Reaction of MBB with the sulfhydryl groups of free cysteines leads to formation of highly fluorescent derivatives. Here we describe a procedure for the detection and relative quantitation of MBB-labeled cysteines using fluorescence and mass spectrometric analyses, which allow determination of free cysteine content and unambiguous identification of MBB-modified cysteine residues. We have applied this approach to the analysis of the free and redox-sensitive cysteine residues of a large membrane protein, the sarcoplasmic reticulum Ca2+ release channel with a molecular mass of 2.2 million Da. Labeling was performed under physiologic conditions where the channel complex is in its native environment and is functionally active. The purified MBB-labeled channel complex was enzymatically digested, and the resulting peptides were separated by reversed phase high-performance chromatography. MBB-labeled peptides were detected by fluorescence and identified by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. Under MALDI conditions, partial photolytic fragmentation of the MBB-peptide bound occurred, thus allowing convenient screening for the MBB-modified peptides in the MS spectrum by detection of the specific mass increment of 190.07 Da for MBB-modified cysteine residues. Modification of the peptides was further confirmed by tandem mass spectrometric analysis, utilizing sequencing information and the presence of the specific immonium ion for the MBB-modified cysteine residues at m/z 266.6. Quantitative information was obtained by comparison of both fluorescence and MS signal intensities of MBB-modified peptides. Combination of fluorescence with MS detection and analysis of MBB-labeled peptides supported by a customized software program provides a convenient method for identifying and quantifying redox sensitive cysteines in membrane proteins of native biological systems. Identification of one redox-sensitive cysteine (2327) in the native membrane bound sarcoplasmic reticulum Ca2+ release channel is described. PMID- 17134129 TI - Identification and quantitative studies of protein immobilization sites by stable isotope labeling and mass spectrometry. AB - A method was developed for characterizing immobilization sites on a protein based on stable isotope labeling and MALDI-TOF mass spectrometry. The model for this work was human serum albumin (HSA) immobilized onto silica by the Schiff base method. The immobilized HSA was digested by various proteolytic enzymes in the presence of normal water, while soluble HSA was digested in (18)O-enriched water for use as an internal standard. These two digests were mixed and analyzed, with the (18)O/(16)O ratio for each detected peptide then being measured. Several peptides in the tryptic, Lys-C, and Glu-C digests gave significantly higher (18)O/(16)O ratios than other peptides in the same digests, implying their involvement in immobilization. Analysis of these results led to identification of the N-terminus and several lysines as likely immobilization sites for HSA (e.g., K4, K41, K190, K225, K313, and K317). It was also possible from these results to quantitatively rank these sites in terms of the relative degree to which each might take part in immobilization. This method is not limited to HSA and silica but can be used with other proteins and supports. PMID- 17134130 TI - Reduction of end effect-induced zone broadening in field-flow fractionation channels. AB - A channel configuration for the elimination of end effects in field-flow fractionation (FFF) channels is simulated and demonstrated for a microfabricated FFF system. In field-flow fractionation, the carrier liquid and sample particles are transferred from a point injection to the full breadth of the rectangular channel using a triangular end piece at the inlet. The nonuniformity in streamline length generated by this end piece results in an increased instrument related plate height. An additional contribution from the end piece at the outlet of the channel further increases the total band broadening. This paper presents a novel approach to minimize end-effect contributions to plate height by fabricating microstructures in the channel end sections to redistribute the flow streams and force streamline lengths to be more uniform. Numerical analysis of the flow profile and sample dispersion (including spreading of particles due to diffusion and advection) is carried out to investigate the optimized microstructure column size, shape, and placement in the end pieces. The configuration obtained from the numerical simulation results is used to design a prototype device. Experimental measurement of the plate heights for this prototype with an on-chip impedance-based detector shows marked improvement in performance due to the presence of the microstructures in comparison to conventional FFF channel geometry with an average 50% reduction in plate height. The redesigned inlet triangle results in a uniform transition of the point injected sample into a thin and straight band across the width of the channel at the start of the rectangular section of the fractionation channel. PMID- 17134131 TI - Methodology utilizing MS signal intensity and LC retention time for quantitative analysis and precursor ion selection in proteomic LC-MALDI analyses. AB - This study describes a methodology for performing relative quantitation in large scale proteomic sample comparisons using an LC-MALDI mass spectrometry analytical platform without the use of isotope tagging reagents. The method utilizes replicate analyses of a sample to create a profile of constituent components that are aligned based on LC elution time and mass. Once components from individual runs have been grouped as common "features", the Student's t test is used to determine which components are systematically different between samples. In this study, five HPLC runs of human plasma were compared to five HPLC runs of human serum. About 3889 components were detected in all 10 runs. Of these, 1831 corresponded to approximately 100 known serum proteins, based on MS/MS analysis of one run each from serum and plasma. As expected, fibrinogen alpha, beta, and gamma chains accounted for many of the most significant differences. Therefore, using MALDI, samples containing thousands of peptides can be compared in a minimal amount of time. Moreover, the results of the comparison can be used to guide further MS/MS mode sample interrogation in a result dependent manner. PMID- 17134132 TI - Multichannel reverse transcription-polymerase chain reaction microdevice for rapid gene expression and biomarker analysis. AB - A microdevice is developed for RNA analysis that integrates one-step reverse transcription and 30 cycles of PCR (RT-PCR) amplification with capillary electrophoresis (CE) separation and fluorescence detection of the amplicons. The four-layer glass-PDMS-glass-glass hybrid microdevice integrates microvalves, on chip heaters and temperature sensors, nanoliter reaction chambers (380 nL), and 5 cm-long CE separation channels. The direct integration of these processes results in attomolar detection sensitivity (<11 template RNA molecules or approximately 0.1 cellular equiv) and rapid 45-min analysis, while minimizing sample waste and eliminating contamination. Size-based electrophoretic product analysis provides definitive amplicon-size verification and multiplex analysis. Multiplexed differential gene expression analysis is demonstrated on mdh and gyrB E. coli transcripts. RNA splice variant analysis of the RBBP8 gene is used to identify tumorigenic tissue. RT-PCR microdevice analysis of normal breast tissue RNA generates the expected 202-bp normal splice isoform; tumor breast tissue RNA samples generate a 151-bp amplicon signifying the presence of the tumorigenic splice variant. The ability to perform RNA transcript and splice variant biomarker analysis establishes our RT-PCR microdevice as a versatile gene expression platform. PMID- 17134133 TI - Separation of acidic and basic proteins by nanoparticle-filled capillary electrophoresis. AB - We present the first example of the analysis of acidic and basic proteins by nanoparticle-filled capillary electrophoresis. Compared to the didodecyldimethylammonium bromide (DDAB)-coated capillary, the DDAB-capped gold nanoparticles (AuNPs) as pseudostationary phase were found to form more stable coating on the capillary wall, thus leading to greater separation efficiency and high reproducibility. In addition to their advantages for protein separation, DDAB-capped AuNPs can generate high reversed electroosmotic flow, which is 75% greater than DDAB at pH 3.5. To allow strong interactions with proteins, the AuNPs were modified with poly(ethylene oxide) via noncovalent bonding to form gold nanoparticles/polymer composites (AuNPPs). Using a capillary dynamically coated with DDAB-capped AuNPs and filled with AuNPPs under acidic conditions (10 mM phosphate, pH 3.5), we have demonstrated the separation of acidic and basic proteins with peak efficiencies ranging from 71 000 to 1 007 000 plates/m and relative standard deviations of migration time less than 0.6%. Additionally, the proposed method has been applied to the analyses of biological samples, including saliva, red blood cells, and plasma. With simplicity, high resolving power, and high reproducibility, the proposed method has shown great potential for proteomics applications and clinical diagnosis. PMID- 17134134 TI - Electrohydrodynamic generation and delivery of monodisperse picoliter droplets using a poly(dimethylsiloxane) microchip. AB - We developed a drop-on-demand microdroplet generator for the discrete dispensing of biosamples into a bioanalytical unit. This disposable PDMS microfluidic device can generate monodisperse droplets of picoliter volume directly out of a plane sidewall of the microfluidic chip by an electrohydrodynamic mechanism. The droplet generation was accomplished without using either an inserted capillary or a monolithically built-in tip. The minimum droplet volume was approximately 4 pL, and the droplet generation was repeatable and stable for at least 30 min, with a typical variation of less than 2.0% of drop size. The Taylor cone, which is usually observed in electrospray, was suppressed by controlling the surface wetting property of the PDMS device as well as the surface tension of the sample liquids. A modification of the channel geometry right before the opening of the microchannel also enhanced the continuous droplet generation without applying any external pumping. A simple numerical simulation of the droplet generation verified the importance of controlling the surface wetting conditions for the droplet formation. Our microdroplet generator can be effectively applied to a direct interface of a microfluidic chip to a biosensing unit, such as AMS, MALDI MS or protein microarray-type biochips. PMID- 17134135 TI - Microwave triggered metal enhanced chemiluminescence: Quantitative protein determination. AB - We present a new technology that offers a faster alternative to the chemiluminescence-based detection that is used in protein assay platforms today. By combining the use of silver nanostructures with chemiluminescent species, a technique that our laboratories have recently shown can enhance the system photon flux over 50-fold, with the use of low-power microwave heating to additionally accelerate, in essence "trigger", chemiluminescence-based reactions, then both ultrafast and ultrabright chemiluminescence assays can be realized. In addition, the preferential heating of the nanostructures by microwaves affords for microwave triggered metal enhanced chemiluminescence (MT-MEC) to be localized in proximity to the silvered surfaces, alleviating unwanted emission from the distal solution. To demonstrate MT-MEC, we have constructed a model assay sensing platform on both silvered and glass surfaces, where comparison with the identical glass substrate-based assay serves to confirm the significant benefits of using silver nanostructures for metal-enhanced chemiluminescence. Our new model assay technology can detect femtomoles of biotinylated BSA in less than 2 min and can indeed be modified to both detect and quantify a great many other biomolecules as well. As compared to traditional western blot approaches, MT-MEC offers protein quantification, high-sensitivity detection combined with ultrafast assay times, i.e., <2 min. PMID- 17134136 TI - Low-power concentration and separation using temperature gradient focusing via Joule heating. AB - We present an experimental study of temperature gradient focusing (TGF) exploiting an inherent Joule heating phenomenon. A simple variable-width PDMS device delivers rapid and repeatable focusing of model analytes using significantly lower power than conventional TGF techniques. High electric potential applied to the device induces a temperature gradient within the microchannel due to the channel's variable width, and the temperature-dependent mobility of the analytes causes focusing at a specific location. The PDMS device also shows simultaneous separation and concentration capability of a mixture of two sample analytes in less than 10 min. An experiment combining Joule heating with external heating/cooling further supports the hypothesis that temperature is indeed the dominant factor in achieving focusing with this technique. PMID- 17134137 TI - In situ visualization of paclitaxel distribution and release by coherent anti Stokes Raman scattering microscopy. AB - Visualization of three-dimensional distribution of drug molecules and subsequent changes during the release process is critical for understanding drug delivery mechanisms as well as designing tailor-made release profiles. This study utilized coherent anti-Stokes Raman scattering (CARS) imaging to examine paclitaxel distribution in various polymer films with lateral resolution of 0.3 microm and depth resolution of 0.9 microm. Raman bands in the CH stretch vibration and fingerprint regions were used to distinguish paclitaxel from the polymers. The detection sensitivity was measured to be 29 mM by imaging paclitaxel molecules dissolved in N,N-dimethylformamide solution. Release of paclitaxel from a polymer matrix was monitored at an acquisition speed of 1 frame/s. Our results show that CARS microscopy can be used effectively for in situ imaging of native drug molecules in a delivery system. PMID- 17134138 TI - Comprehensive study of lead pollution in Shanghai by multiple techniques. AB - Aerosol samples of PM10 particulates were examined with particle-induced X-ray emission (PIXE and micro-PIXE ), inductively coupled plasma-mass spectrometry, and X-ray absorption fine structures (XAFS) to investigate atmospheric Pb concentrations and its chemical species and to decide the source assignment. From the elemental analyses, lead concentrations were averaged at 369 and 237 ng.m-3 in aerosol samples of PM10 collected from 19 monitor sites in Shanghai in the winter of 2002 and 2003, respectively. The XAFS results show that major chemical forms of the lead particulates are probably PbCl2, PbSO4, and PbO. A calculation of isotope ratio and chemical mass balance of the PM10 samples revealed that the atmospheric lead particulates in Shanghai in the two periods were mainly from coal combustors, iron and steel plants, and automobile exhausts, with coal combustion dominating the lead pollution after the phasing out of leaded gasoline in 1997 in Shanghai. PMID- 17134139 TI - Second-order advantage achieved by unfolded-partial least-squares/residual bilinearization modeling of excitation-emission fluorescence data presenting inner filter effects. AB - A second-order multivariate calibration approach, based on a combination of unfolded-partial least-squares with residual bilinearization (U-PLS/RBL), has been applied to fluorescence excitation-emission matrix data for multicomponent mixtures showing inner filter effects. The employed chemometric algorithm is the most successful one regarding the prediction of analyte concentrations when significant inner filter effects occur, even in the presence of unexpected sample components, which require strict adherence to the second-order advantage. Results for simulated fluorescence excitation-emission data are described, in comparison with the classical approach based on parallel factor analysis and other second order algorithms, including generalized rank annihilation, bilinear least squares combined with residual bilinearization and multivariate curve resolution alternating leastsquares. A set of experimental data was also studied, in which calibration was performed with fluorescence excitation-emission matrices for samples containing mixtures of chrysene (the analyte of interest) and benzopyrene (which produced strong inner filter effect across the useful wavelength range). Prediction was made on validation samples with a qualitative composition similar to the calibration set, and also on test samples containing an unexpected component (pyrene). In this latter case, U-PLS/RBL showed a unique success for the analysis of the calibrated component chrysene, achieving the useful second order advantage. PMID- 17134140 TI - Collision-induced dissociative chemical cross-linking reagents and methodology: Applications to protein structural characterization using tandem mass spectrometry analysis. AB - Chemical cross-linking combined with mass spectrometry is a viable approach to study the low-resolution structure of protein and protein complexes. However, unambiguous identification of the residues involved in a cross-link remains analytically challenging. To enable a more effective analysis across various MS platforms, we have developed a novel set of collision-induced dissociative cross linking reagents and methodology for chemical cross-linking experiments using tandem mass spectrometry (CID-CXL-MS/MS). These reagents incorporate a single gas phase cleavable bond within their linker region that can be selectively fragmented within the in-source region of the mass spectrometer, enabling independent MS/MS analysis for each peptide. Initial design concepts were characterized using a synthesized cross-linked peptide complex. Following verification and subsequent optimization of cross-linked peptide complex dissociation, our reagents were applied to homodimeric glutathione S-transferase and monomeric bovine serum albumin. Cross-linked residues identified by our CID CXL-MS/MS method were in agreement with published crystal structures and previous cross-linking studies using conventional approaches. Common LC/MS/MS acquisition approaches such as data-dependent acquisition experiments using ion trap mass spectrometers and product ion spectral analysis using SEQUEST were shown to be compatible with our CID-CXL-MS/MS reagents, obviating the requirement for high resolution and high mass accuracy measurements to identify both intra- and interpeptide cross-links. PMID- 17134141 TI - Simultaneous dielectrophoretic separation and assembly of single-walled carbon nanotubes on multigap nanoelectrodes and their thermal sensing properties. AB - By using the specifically designed multigap nanoelectrodes, we demonstrated an effective approach for the simultaneous dielectrophoretic separation and assembly of metallic and semiconducting single-walled carbon nanotubes (SWNTs). An approximate metallic-semiconducting-metallic multiarray structure was created by an inward-propagative sequential assembly of SWNTs under ac electric field. Such kinds of SWNT multiarray structures exhibited ultra-low-power consumption and excellent thermal sensing performances with the sensitivity being dependent on the number of gaps: the more gaps, the higher sensitivity. The effective separation of metallic and semiconducting tubes in different gaps is believed to be responsible for the improved sensitivity to temperature. PMID- 17134142 TI - Genetically encoded optical probe for detecting release of proteins from mitochondria toward cytosol in living cells and mammals. AB - We developed a genetically encoded bioluminescence indicator for monitoring the release of proteins from the mitochondria in living cells. The principle of this method is based on reconstitution of split Renilla reniformis luciferase (Rluc) fragments by protein splicing with an Ssp DnaE intein. A target mitochondrial protein connected with an N-terminal fragment of Rluc and an N-terminal fragment of DnaE is expressed in mammalian cells. If the target protein is released from the mitochondria toward the cytosol upon stimulation with a specific chemical, the N-terminal Rluc meets the C-terminal Rluc connected with C-terminal DnaE in the cytosol, and thereby, the full-length Rluc is reconstituted by protein splicing. The extent of release of the target fusion protein is evaluated by measuring activities of the reconstituted Rluc. To test the feasibility of this method, here we monitored the release of Smac/DIABLO protein from mitochondria during apoptosis in living cells and mice. The present method allowed high throughput screening of an apoptosis-inducing reagent, staurosporine, and imaging of the Smac/DIABLO release in cells and in living mice. This rapid analysis can be used for screening and assaying chemicals that would increase or inhibit the release of mitochondrial proteins in living cells and animals. PMID- 17134144 TI - Generating multiple independent retention index data in dual-secondary column comprehensive two-dimensional gas chromatography. AB - A method producing simultaneously three retention indexes for compounds has been developed for comprehensive two-dimensional gas chromatography by using a dual secondary column approach (GC x 2GC). For this purpose, the primary flow of the first dimension column was equally diverted into two secondary microbore columns of identical geometry by means of a three-way flow splitter positioned after the longitudinally modulated cryogenic system. This configuration produced a pair of comprehensive two-dimensional chromatograms and generated retention data on three different stationary phases in a single run. First dimension retention indexes were determined on a polar SolGel-Wax column under linear programmed-temperature conditions according to the van den Dool approach using primary alcohol homologues as the reference scale. Calculation of pseudoisothermal retention indexes in both second dimensions was performed on low-polarity 5% phenyl equivalent polysilphenylene/siloxane (BPX5) and 14% cyanopropylphenyl/86% dimethylpolysiloxane (BP10) columns. To construct a retention correlation map in the second dimension separation space upon which KovAts indexes can be derived, two methods exploiting "isovolatility" relationships of alkanes were developed. The first involved 15 sequential headspace samplings of selected n-alkanes by solid-phase microextraction (SPME), with each sampling followed by their injection into the GC at predetermined times during the chromatographic run. The second method extended the second dimension retention map and consisted of repetitive introduction of SPME-sampled alkane mixtures at various isothermal conditions incremented over the temperature program range. Calculated second dimension retention indexes were compared with experimental values obtained in conventional one-dimensional GC. A case study mixture including 24 suspected allergens (i.e., fragrance ingredients) was used to demonstrate the feasibility and potential of retention index information in comprehensive 2D-GC. PMID- 17134143 TI - De novo sequencing and disulfide mapping of a bromotryptophan-containing conotoxin by Fourier transform ion cyclotron resonance mass spectrometry. AB - T-1-family conotoxins belong to the T-superfamily and are composed of 10-17 amino acids. They share a common cysteine framework and disulfide connectivity and exhibit unusual posttranslational modifications, such as tryptophan bromination, glutamic acid carboxylation, and threonine glycosylation. We have isolated and characterized a novel peptide, Mo1274, containing 11 amino acids, that shows the same cysteine pattern, -CC-CC, and disulfide linkage as those of the T-1-family members. The complete sequence, GNWCCSARVCC, in which W denotes bromotryptophan, was derived from MS-based de novo sequencing. The FT-ICR MS/MS techniques of electron capture dissociation (ECD), infrared multiphoton dissociation, and collision-induced dissociation served to detect and localize the tryptophan bromination. The bromine contributes a distinctive isotopic distribution in all fragments that contain bromotryptophan. ECD fragmentation results in the loss of bromine and return to the normal isotopic distribution. Disulfide connectivity of Mo1274, between cysteine pairs 1-3 and 2-4, was determined by mass spectrometry in combination with chemical derivatization employing tris(2 carboxyethyl)phosphine, followed by differential alkylation with N-ethylmaleimide and iodoacetamide. The ECD spectra of the native and partially modified peptide reveal a loss of bromine in a process that requires the presence of a disulfide bond. PMID- 17134145 TI - Determination of the concentration of single-walled carbon nanotubes in aqueous dispersions using UV-visible absorption spectroscopy. AB - Stable, homogeneous, aqueous dispersions of single-walled carbon nanotubes (SWNTs) are prepared by nonspecific physical adsorption of surfactants enhanced by sonication. Upon centrifugation, supernatant and precipitate phases are obtained. The initial weights of the SWNTs and the surfactant are divided between these two phases, and the respective SWNT concentration in each phase is unknown. The focus of this work is on the determination of the true concentration of raw, exfoliated HiPCO SWNTs in the supernatant phase. A UV-visible absorption-based approach is suggested for a direct measurement of the SWNT and the surfactant concentration in the supernatant. UV-visible absorbance spectra of SWNTs surfactant dispersions and surfactants alone reveal that the intensity of a certain peak, attributed to the pi-plasmon resonance absorption, is unaffected by the presence of most surfactants. A calibration plot is then made by monitoring the intensity of the peak as a function of the true concentration of the exfoliated SWNTs. Thus, we are able to determine the unknown concentration of surfactant-dispersed HiPCO SWNTs in the supernatant solution, simply by measuring its optical absorbance. Moreover, we can now calculate the surfactant efficiency in dispersing SWNTs. Cryogenic-transmission electron microscopy and thermogravimetric analysis techniques are used for the characterization of these dispersions and to complement the UV-visible measurements. PMID- 17134146 TI - Partial validation of cross flow ultrafiltration by atomic force microscopy. AB - Atomic force microscopy was used to evaluate a cross flow ultrafiltration (CFUF) system. The CFUF system was used for the size fractionation of natural colloidal material from freshwaters. Analysis of the images of bulk water, permeates, and retentates shows the primary materials observed were near-spherical structures with height dimensions up to approximately 12 nm. The number of colloidal particles (per unit area) on the mica surfaces derived from the retentates increased by a factor of 2 between a concentration factor (cf) of 1 and of 20. Colloidal densities of nanoparticles were approximately 2 orders of magnitude lower in the permeate compared to the retentate, indicating a good size fractionation. As the cf value increased from 1 to 20, the percentage of <1-nm material decreased substantially and the percentage of >1-nm material increased substantially in the retentates. Line transects along a surface and surface roughness values show good agreement with the above results. Results suggest the size fractionation is good but not perfect and high cf values produce a better size fractionation, although some retention of small material is always observed. High cf values are recommended. PMID- 17134147 TI - Uncertainty-based internal quality control. Harmonization considerations. AB - Three main quality aspects for analytical laboratories are internal method validation, internal quality control (IQC), and sample result uncertainty. Unfortunately, in the past they have been used in a nonharmonized way. The most universal IQC tool is the mean chart, but some criteria used to fix their control limits do not fit the real nature of analytical results. A new approach for fixing these limits is proposed (the u-approach). The key is the combined uncertainty, u, obtained from the method validation information, also used for estimating the sample result uncertainty. A comparative study on "in-control" simulated, bibliographic, and real laboratory data suggests that the u-approach is more reliable than other well-established criteria. In addition, the u approach mean chart emerges as an IQC tool, consistent with chemical assays, which harmonizes the validation-control-uncertainty process. PMID- 17134148 TI - Assay of trace D-amino acids in neural tissue samples by capillary liquid chromatography/tandem mass spectrometry. AB - A sensitive chiral capillary HPLC-MS/MS method well suited for the determination of amino acid enantiomers in biological samples was developed. The method involved precolumn derivatization of the sample with 7-fluoro-4 nitrobenzoxadiazole (NBD-F). After derivatization, NBD-amino acids were stacked on a C18 reversed-phase extraction microcolumn, thus enriching and cleaning up the analytes. Various chiral stationary phases (CSPs) including cyclodextrin bonded silica, Pirkle-type, vancomycin, and teicoplanin-bonded silica particles were evaluated for resolving NBD-F tagged amino acid enantiomers with mobile phases compatible with MS detection. It was found that only teicoplanin aglycon CSP provided sufficient resolution of NBD-Asp and NBD-Ser enantiomers to quantify trace levels of D-Asp and D-Ser in tissue samples. MS/MS detection of NBD-amino acid derivatives was very sensitive and selective. The high selectivity allowed the use of a stable isotope-labeled analyte analogue (i.e., L-aspartic acid-2,3,3 d3) as internal standard for the quantitation to improve assay reproducibility and reliability. Neural tissue samples dissected from rat brain and the central nervous system (CNS) of Aplysia californica, a widely used neuronal model, were analyzed to determine the chirality of glutamic acid (Glu), aspartic acid (Asp), and serine (Ser). The former two are major excitatory amino acids in the brain, and the last one has been recently identified as a neuromodulator. Both D-Ser and D-Asp were detected in rat brain. While the D-Asp level decreased rapidly through the developmental stages of the rat, the D-Ser level increased steadily from 82.3 microg/g of wet tissue in 3-day prenatal rats to 241.3 microg/g of wet tissue in 90-day-old rats. Interestingly, no D-Ser was detected in the CNS of Aplysia, a "primitive" invertebrate. However, the D-Asp level in this animal was found to be high. In a particular connective nerve sample, D-Asp was at 323.2 microg/g of wet tissue and constituted 60.2% of total Asp. D-Glu was not detected either in rat brain or in Aplysia's CNS. PMID- 17134149 TI - Differential pulse voltammetry and additive differential pulse voltammetry with solvent polymeric membrane ion sensors. AB - The ion transfer across the water-solvent polymeric membrane interface is investigated by using a new device based on a modification of a commercial ion selective electrode body that permits the accommodation of a platinum counter electrode inside the inner filling solution compartment and, therefore, use of a four-electrode potentiostat with ohmic drop compensation. This device is used here to apply two different double potential pulse techniques--differential pulse voltammetry and additive differential pulse voltammetry--which are more advantageous than other voltammetric techniques, such as normal pulse voltammetry or cyclic voltammetry, for the determination of the characteristic electrochemical parameters of the system. This is due to the concurrence of two factors in these double potential pulse techniques, the peak-shaped response together with a considerable reduction of undesirable current contributions. PMID- 17134150 TI - Charged species transport, separation, and dispersion in nanoscale channels: autogenous electric field-flow fractionation. AB - Numerical methods are employed to examine the transport of charged species in pressure-driven and electroosmotic flow along nanoscale channels having an electric double-layer thickness comparable to the channel size. In such channels, the electric field inherent to the double layer produces transverse species distributions that depend on species charge. Flow along the channel thus yields mean axial species speeds that also depend on the species charge, enabling species separation and identification. Here we characterize field-flow separations of this type via the retention and plate height. For pressure-driven flows, we demonstrate that mean species speeds along the channel are uniquely associated with a single species charge, allowing species separation based on charge alone. In contrast, electroosmotic flows generally yield identical speeds for several values of the charge, and these speeds generally depend on both the species charge and electrophoretic mobility. Coefficients of dispersion for charged species in both planar and cylindrical geometries are presented as part of this analysis. PMID- 17134151 TI - Surfactant gradient methods using mixed systems of cethyltrimethylammonium chloride and nonionic surfactants possessing polyoxyethylene chains for electrokinetic separation of benzoate anions as model analytes. AB - Surfactant gradient methods for electrokinetic separation of 10 benzoates as model organic anions were investigated using mixed micellar solutions of cetyltrimethylammonium chloride (CTAC) and nonionic surfactants possessing polyoxyethylene chains, polyoxyethylene sorbitan monolaurate (Tween 20) or polyoxyethylene lauryl ether (Brij 35). Electroosmotic flow (EOF) was eliminated virtually by a coating of the inner wall of the capillaries, and then the benzoates were detected fundamentally in the order of their hydrophobicity. In a pure CTAC system, the synergistic influences of attractive electrostatic and hydrophobic interactions gave rise to quite large retention factors of many of the benzoate anions, resulting in their coelution. Addition of an adequate amount of Tween 20 to the pure CTAC system decreased the electrostatic interaction significantly to give remarkably improved separation of the analytes, but long analysis time was required. A surfactant gradient method would be useful to decrease analysis time and to improve separation simultaneously. Under slight EOF, the micelles in the inlet reservoir can pass through and, thus, interact with all of the analytes before they were detected. In the present system, surfactant gradient separations could be performed simply by changing compositions of the surfactant solutions in the inlet reservoir during a single run. Additionally, we carried out continuous gradient separation using a simple device. Brij 35 gave an effect parallel to that by Tween 20 in migration behavior of the analytes. A practically negligible change in the level of the baseline was observed in a stepwise gradient elution with the CTAC/Brij 35 system because of the small absorbance at the detection wavelength, while that with the CTAC/Tween 20 was considerable. All the benzoates were separated completely within reasonable analysis times using both stepwise and continuous gradient programs for the concentrations of Tween 20 or Brij 35 in the presence of 100 mM CTAC. PMID- 17134152 TI - Determination of peroxide-based explosives using liquid chromatography with on line infrared detection. AB - A nondestructive analytical method for peroxide-based explosives determination in solid samples is described. Reversed-phase high-performance liquid chromatography in combination with on-line Fourier transform infrared (FT-IR) detection is used for the analysis of triacetonetriperoxide (TATP) and hexamethylenetriperoxide diamine (HMTD). In contrast to other liquid chromatographic methods with optical detection, no derivatization or decomposition of the peroxides is required. The peroxides are identified and quantified via their characteristic absorption spectra in the mid-infrared range of the electromagnetic spectrum. The detection limit of 0.5 mmol L-1 for HMTD and 1 mmol L-1 for TATP allows the identification of the explosives in complex matrixes. PMID- 17134153 TI - Structure elucidation of an artifact discharging from rubber-based vial closures by means of gas chromatography/tandem mass spectrometry. AB - The use of vial closures equipped with butyl rubber septa may lead to sample contamination by rubber additives discharging from the septum material. In this study, the structure elucidation of an artifact causing intense signals in gas chromatography/electron capture negative ion mass spectrometry (GC/ECNI-MS) and gas chromatographic analyses with electron capture detection is described. Tentative identification of the leached compound was achieved by employing tandem mass spectrometric techniques both in electron capture negative ion and in electron ionization modes. The artifact could thus be characterized as 2 benzothiazolyl-N,N-dimethyl dithiocarbamate, which is a known vulcanization accelerator for rubber. It is conceivable that the identified compound or related substances are also used in other applications. Therefore, two food-related matrixes were investigated for a possible migration of this compound into foods. During these analyses, the tentatively identified rubber additive was detected in an aqueous extract of a rubber seal ring for canning jars. GC/ECNI-MS provided better sensitivity and selectivity than GC/EI-MS for the determination of the rubber additive and other mercaptobenzothiazole-derived substances. PMID- 17134154 TI - Atmospheric pressure photoionization for ionization of both polar and nonpolar compounds in reversed-phase LC/MS. AB - Atmospheric pressure photoionization can provide high ionization efficiency simultaneously to both polar and nonpolar compounds delivered in reversed-phase solvent. The method to achieve this utilizes toluene as a dopant and simply requires that the solvent flow be limited so that reactions between toluene photoions and the organic component of the solvent are not driven to completion. Under these conditions, toluene photoions remain in the source for ionizing nonpolar compounds via charge exchange (electron transfer), while protonated solvent ions are available for proton-transfer reactions with polar molecules. The reagent ion mixture is then suitable for ionizing a wide range of both polar and nonpolar compounds. The critical effect of solvent flow rate is demonstrated here with results for a test analyte, 9-methylanthracene, which may be ionized by either charge exchange or proton transfer. For a solvent of 50:50 methanol/water (v/v), lowering the flow from 200 to 50 microL min-1 results in a 10x increase in charge exchange ionization efficiency--further flow reductions provide even greater enhancements. This method is compatible with sample delivery by direct infusion and micro- and narrow-bore LC, as well as conventional LC using a flow splitter. PMID- 17134155 TI - Method to improve linearity of diffuse reflection mid-infrared spectroscopy. PMID- 17134156 TI - Combined genetic algorithm and multiple linear regression (GA-MLR) optimizer: Application to multi-exponential fluorescence decay surface. AB - The optimization approach based on the genetic algorithm (GA) combined with multiple linear regression (MLR) method, is discussed. The GA-MLR optimizer is designed for the nonlinear least-squares problems in which the model functions are linear combinations of nonlinear functions. GA optimizes the nonlinear parameters, and the linear parameters are calculated from MLR. GA-MLR is an intuitive optimization approach and it exploits all advantages of the genetic algorithm technique. This optimization method results from an appropriate combination of two well-known optimization methods. The MLR method is embedded in the GA optimizer and linear and nonlinear model parameters are optimized in parallel. The MLR method is the only one strictly mathematical "tool" involved in GA-MLR. The GA-MLR approach simplifies and accelerates considerably the optimization process because the linear parameters are not the fitted ones. Its properties are exemplified by the analysis of the kinetic biexponential fluorescence decay surface corresponding to a two-excited-state interconversion process. A short discussion of the variable projection (VP) algorithm, designed for the same class of the optimization problems, is presented. VP is a very advanced mathematical formalism that involves the methods of nonlinear functionals, algebra of linear projectors, and the formalism of Frechet derivatives and pseudo-inverses. Additional explanatory comments are added on the application of recently introduced the GA-NR optimizer to simultaneous recovery of linear and weakly nonlinear parameters occurring in the same optimization problem together with nonlinear parameters. The GA-NR optimizer combines the GA method with the NR method, in which the minimum-value condition for the quadratic approximation to chi(2), obtained from the Taylor series expansion of chi(2), is recovered by means of the Newton-Raphson algorithm. The application of the GA-NR optimizer to model functions which are multi-linear combinations of nonlinear functions, is indicated. The VP algorithm does not distinguish the weakly nonlinear parameters from the nonlinear ones and it does not apply to the model functions which are multi-linear combinations of nonlinear functions. PMID- 17134157 TI - Conformational analysis. 24. Structure and composition of gaseous oxalyl fluoride, C(2)F(2)O(2): electron-diffraction investigation augmented by data from microwave spectroscopy and molecular orbital calculations. AB - The molecular structure and composition of gaseous oxalyl fluoride (OXF) has been investigated by electron diffraction (GED) at nozzle-tip temperatures of -10, 149, and 219 degrees C. The GED data were augmented by molecular orbital calculations, and the analysis was aided by use of rotational constants from microwave (MW) spectroscopy. As in the other oxalyl halides, there are two stable species, of which the more stable is periplanar anti (i.e., trans). However, unlike these other halides in which the second form is gauche, the second form of oxalyl fluoride was known from MW work to be periplanar syn (i.e., cis). Our results are consistent with a mixture of trans and cis forms, and yield values for the structural parameters, the composition of the system at the three temperatures cited, and the thermodynamic quantities deltaG(o), deltaH(o), and deltaS(o) for the reaction trans --> cis. Some trans/cis distances (r(g)/Angstrom) and angles (<(alpha)/deg) at -10 degrees C are r(C=O) = 1.178(2)/1.176(2), r(C-F) = 1.323(2)/1.328(2); r(C-C) = 1.533(3)/1.535(3), <(C C=O) = 126.4(2)/124.2(2), <(C-C-F) = 109.8(2)/112.2(2), and <(O-C-F) = 123.8(2)/123.6(2). The mixture compositions (percent trans) at -10 degrees C/149 degrees C/219 degrees C are 75(3)/58(7)/52(8), from which deltaH(o) and deltaSO) are found to be 1.14 kcal/mol and 2.12 cal/(mol x deg). The system properties are discussed. PMID- 17134158 TI - IR spectrum of the H(5)O(2)(+) cation in the context of proton disolvates L-H(+) L. AB - The H(5)O(2)(+) ion has been studied in chlorocarbon, benzene, and weakly coordinating anion environments to bridge the gap between the gas-phase and traditional condensed-phase investigations. Symmetrical cations of the type [H(5)O(2)(+) x 4Solv] are formed via H-bonding with the terminal O-H groups. In the infrared spectrum, the nu(s)OH and nu(as)OH vibrations behave in a manner similar to those of common water molecules: the stronger is the H-bonding interaction with the surroundings, the lower is the frequency shift. A consistent pattern of IR bands from the central O-H(+)-O group is identified, regardless of the strength of the interaction of H(5)O(2)(+) with its environment. Three intense bands develop: a (860-995 cm-1), b (1045-1101 cm(-1)), and c (1672-1700 cm(-1)), as well as two weak bands, d ( approximately 1300 cm(-1)) and e ( approximately 1400-1500 cm(-1)). These fingerprint bands are highly characteristic for vibrations of O-H-O group irrespective of formal charge. They are seen in symmetrical proton disolvates of the type L-H(+)-L, where L is an O atom donor (alcohol, ether, ketone, phosphate, etc.), and in [A-H-A](-) acid salts (A(-) = oxyanion). The commonality is equivalency of the two O-atoms, a short O...O distance (ca. 2.40 Angstrom), and a flat-bottomed potential well for the bridging proton, that is, a short, strong, low-barrier H-bond. Assignments for bands a-e are suggested in an attempt to resolve inconsistencies between experimental and calculated data. PMID- 17134159 TI - Toroidal hopping of a single hole through the circularly-arrayed naphthyl groups in hexanaphthylbenzene cation radical. AB - Synthesis of a dendritic (soluble) hexanaphthylbenzene derivative is described in which the six naphthyl groups are connected to the central benzene ring in a propeller-shaped arrangement. Observation of multiple oxidation waves in its cyclic voltammogram as well as an intense charge-resonance transition (extending beyond 1600 nm) in its cation radical, generated by laser-flash photolysis using photoexcited chloranil as an oxidant, suggests that a single hole is mobilized via electron transfer (or hopping) over six identical (circularly arrayed) redox centers. PMID- 17134160 TI - Investigation of the UV/visible absorption spectra of merocyanine dyes using time dependent density functional theory. AB - The visible spectral properties of a set of merocyanine dyes have been determined experimentally and compared to theoretical estimates calculated using time dependent density functional theory. Three families of merocyanines have been investigated. The merocyanines of the first class contain the 4-thiazolidinone, 2 thioxo group in resonance with different donor groups. In the second family, they present a common pyrrolo[2,3-c]pyrazole-6-(2H)-acetic acid, the 4-[4 (dialkylamino)phenyl]-3,5-dihydro-3,5-dioxo-2-phenyl group with different substituents on the two phenyl rings, while in the third family, they are constituted of an isoxazolone group linked via a conjugated polyenic segment to an aminophenyl group. Two hybrid exchange-correlation functionals have been used, whereas solvent effects have been included using the integral equation formalism of the polarizable continuum model. Moreover, for one compound, the vibrational structure of the electronic transitions has been determined by calculating the Franck-Condon factors within the Born-Oppenheimer approximation. The calculations show that the dominant transitions present a pi-pi(*) character. This approach reproduces nicely the variations of excitation energies with the nature of the merocyanine so that least-squares fits can be used to correct for the major systematic errors of the computational scheme and to reach an accuracy of 0.09 eV or better. Part of these systematic errors is shown to originate from the description of the solvent effects, whereas vibronic effects can account for most of the remaining differences. PMID- 17134161 TI - Fingerprints of delocalized transition states in quantum dynamics. AB - Reactions with delocalized transition states (plateau reactions) can be characterized statically by their energy profile along the reaction path, where they exhibit a broad, flat region instead of one or several well-defined saddle points on the potential energy surface. Employing our new, highly flexible quantum dynamics code to perform two-dimensional and effective four-dimensional quantum wave packet propagations on ab initio based model potentials, we show that plateau reactions can also be discerned from the other standard reaction types by their dynamics. PMID- 17134162 TI - Temperature-dependent complex indices of refraction for crystalline (NH(4))(2)SO(4). AB - We present a significantly improved set of complex indices of refraction (optical constants) for crystalline (NH(4))(2)SO(4) at 298 K, determined from extinction spectra measured in an aerosol flow tube (AFT). The improved values provide more accurate reproductions of experimental extinction spectra when used in light scattering calculations (Mie, T-matrix, etc.). Optical constants were also derived from measurements using a cryogenic AFT at 243, 223, and 213 K, temperatures characteristic of the upper troposphere and stratosphere. Only minor changes in the optical constants were observed down to 223 K, the transition temperature to the ferroelectric phase, after which significant changes were observed. Here we report the first complex indices of refraction at reduced temperatures for both phases of crystalline (NH(4))(2)SO(4). PMID- 17134163 TI - Kinetic and product study of the gas-phase reactions of OH radicals, NO(3) radicals, and O(3) with (C(2)H(5)O)(2)P(S)CH(3) and (C(2)H(5)O)(3)PS. AB - Rate constants for the reactions of OH radicals and NO3 radicals with O,O-diethyl methylphosphonothioate [(C(2)H(5)O)(2)P(S)CH(3); DEMPT] and O,O,O-triethyl phosphorothioate [(C(2)H(5)O)(3)PS; TEPT] have been measured using relative rate methods at atmospheric pressure of air over the temperature range 296-348 K for the OH radical reactions and at 296 +/- 2 K for the NO(3) radical reactions. At 296 +/- 2 K, the rate constants obtained for the OH radical reactions (in units of 10(-11) cm(3) molecule(-1) s(-1)) were 20.4 +/- 0.8 and 7.92 +/- 0.27 for DEMPT and TEPT, respectively, and those for the NO(3) radical reactions (in units of 10(-15) cm(3) molecule(-1) s(-1)) were 2.01 +/- 0.20 and 1.03 +/- 0.10, respectively. Upper limits to the rate constants for the reactions of O(3) with DEMPT and TEPT of <6 x 10(-20) cm(3) molecule(-1) s(-1) were determined in each case. Rate constants for the OH radical reactions, measured relative to k(OH + alpha-pinene) = 1.21 x 10(-11) e(436/T) cm(3) molecule(-1) s(-1), resulted in the Arrhenius expressions k(OH + DEMPT) = 1.08 x 10(-11) e(871+/-25)/T cm(3) molecule(-1) s(-1) and k(OH + TEPT) = 8.21 x 10(-13) e(1353+/-49)/T cm(3) molecule(-1) s(-1) over the temperature range 296-348 K, where the indicated errors are two least-squares standard deviations and do not include the uncertainties in the reference rate constant. Diethyl methylphosphonate was identified and quantified from the OH radical and NO(3) radical reactions with DEMPT, with formation yields of 21 +/- 4%, independent of temperature, from the OH radical reaction and 62 +/- 11% from the NO(3) radical reaction at 296 +/- 2 K. Similarly, triethyl phosphate was identified and quantified from the OH radical and NO(3) radical reactions with TEPT, with formation yields of 56 +/- 9%, independent of temperature, from the OH radical reaction and 78 +/- 15% from the NO(3) radical reaction at 296 +/- 2 K. PMID- 17134164 TI - Theoretical study on the optoelectronic properties of electron-withdrawing substituted diethynylfluorenyl gold(I) complexes. AB - We report on a quantum-chemical study of the electronic and optical properties of gold(I) complex AuTFT (1) and its electron-withdrawing substitutents, AuTFOT (2) and AuTFCNT (3) [where TFT = diethynylfluorenyl, TFOT = diethynylfluorenone and TFCNT = diethynyl-(9-(dicyanomethylene)fluorene)]. Our theoretical calculations indicate that for all systems the reorganization energies of electron and hole are in the same order of magnitude and similar to those of the well-known electrontransport material Alq3. The substitution of -CO and -C(CN)2 for -CH in AuTFT significantly decreases the bond length alternation and increases the electron affinity, which would effectively lower the energy barrier for electron injection from cathode and thus qualify AuTFOT (2) and AuTFCNT (3) as candidates for the electron transport layer (ETL) in light-emitting diodes (LEDs). The lowest lying excited-states of gold(I) diethynylfluorenyl derivatives have been studied by the singles configuration interaction (CIS) method and time-dependent density functional method (TDDFT). It is found that the electron-withdrawing substitutions evidently decrease the energy gap, leading to a remarkable red shift in transition energy and transformation in the direction of charge transfer. Our research is important in the development of new functional materials for the design of LEDs with enhanced performance. PMID- 17134165 TI - Excited-state ab initio calculations and multidimensional Franck-Condon simulations on guanine. AB - The guanine enol and keto N7H and N9H tautomers have been optimized at the CASSCF/cc-pVDZ levels of theory. Except for the enol N7H tautomer, CASSCF predicts distorted nonplanar S1 state geometries. Among the vibronic simulations carried out with the optimized structures only the enol N7H tautomer qualitatively mirrors the appearance of the experimental R2PI spectrum. Refined symmetry-adapted cluster configuration interaction (SACCI) geometries of the enol N7H tautomer produce simulations in good agreement with experiment and support the assignment of the first vibronic band and associated vibronic features of the R2PI spectrum to this tautomer. The sharp spectral features and the fact that Franck-Condon simulations based on the harmonic approximation allow for a faithful reproduction of the spectral signature associated with the enol N7H tautomer indicate that within the simulated energy window the S1 potential energy surface of this isomer is fairly harmonic and free from conical intersections involved in the S1 state lifetime-shortening relaxation processes of other DNA bases and possibly the remaining tautomers of guanine. PMID- 17134166 TI - Enthalpies of formation, bond dissociation energies, and molecular structures of the n-aldehydes (acetaldehyde, propanal, butanal, pentanal, hexanal, and heptanal) and their radicals. AB - Aldehydes are important intermediates and products in a variety of combustion and gas-phase oxidation processes, such as in low-temperature combustion, in the atmosphere, and in interstellar media. Despite their importance, the enthalpies of formation and bond dissociation energies (BDEs) for the aldehydes are not accurately known. We have determined enthalpies of formation for acetaldehyde, propanal, and butanal from thermodynamic cycles, using experimentally measured reaction and formation enthalpies. All enthalpy values used for reference molecules and reactions were first verified to be accurate to within around 1 kcal mol-1 using high-level ab initio calculations. Enthalpies of formation were found to be -39.72 +/- 0.16 kcal mol-1 for acetaldehyde, -45.18 +/- 1.1 kcal mol 1 for propanal, and -49.27 +/- 0.16 kcal mol-1 for butanal. Enthalpies of formation for these three aldehydes, as well as for pentanal, hexanal, and heptanal, were calculated using the G3, G3B3, and CBS-APNO theoretical methods, in conjunction with bond-isodesmic work reactions. On the basis of the results of our thermodynamic cycles, theoretical calculations using isodesmic work reactions, and existing experimental measurements, we suggest that the best available formation enthalpies for the aldehydes acetaldehyde, propanal, butanal, pentanal, hexanal, and heptanal are -39.72, -45.18, -50.0, -54.61, -59.37, and 64.2 kcal mol-1, respectively. Our calculations also identify that the literature enthalpy of formation of crotonaldehyde is in error by as much as 1 kcal mol-1, and we suggest a value of -25.1 kcal mol-1, which we calculate using isodesmic work reactions. Bond energies for each of the bonds in the aldehydes up to pentanal were calculated at the CBS-APNO level. Analysis of the BDEs reveals the R-CH(2)CH=O to be the weakest bond in all aldehydes larger than acetaldehyde, due to formation of the resonantly stabilized vinoxy radical (vinyloxy radical/formyl methyl radical). It is proposed that the vinoxy radical as well as the more commonly considered formyl and acetyl radicals are important products of aldehyde combustion and oxidation, and the reaction pathways of the vinoxy, formyl, and acetyl radicals are discussed. Group additivity values for the carbon-oxygen hydrogen groups common to the aldehydes are also determined. Internal rotor profiles and electrostatic potential surfaces are used to study the dipole induced dipole-dipole interaction in the synperiplanar conformation of propanal. It is proposed that the loss of this dipole-dipole interaction in RC(. )HCH(2)CH=O radicals causes a ca. 1-2 kcal mol-1 decrease in the aldehyde C-H and C-C bond energies corresponding to RC(.-)HCH(2)CH=O radical formation. PMID- 17134167 TI - Theoretical study of the antioxidant properties of pyridoxine. AB - Molecules acting as antioxidants capable of scavenging reactive oxygen species (ROS) are of utmost importance in the living cell. The antioxidative properties of pyridoxine (vitamin B6) have recently been discovered. In this study, we have analyzed the reactivity of pyridoxine toward the ROS (.-)OH, (.-)OOH, and (. )O(2)- at the density functional theory level (functionals B3LYP and MPW1B95). Two reaction types have been studied as follows: addition to the aromatic ring atoms and hydrogen/proton abstraction. Our results show that (.-)OH is the most reactive species, while (.-)OOH displays low reactivity and (.-)O2(-) does not react at all with pyridoxine. The most exergonic reactions are those where (.-)H is removed from the CH(2)OH groups or the ring-bound OH group and range from -33 to -39 kcal/mol. The most exergonic addition reactions occur by attacking the carbon atoms bonded to nitrogen but with an energy gain of only 6 kcal/mol. PMID- 17134168 TI - Molecular electrostatic potentials and hydrogen bonding in alpha-, beta-, and gamma-cyclodextrins. AB - Cyclodextrins (CDs) are cyclic oligomers of glucose having the toroid of sugars elaborating a central cavity of varying size depending on the number of glucoses. The central hydrophobic cavity of CD shows a binding affinity toward different guest molecules, which include small substituted benzenes to long chain surfactant molecules leading to a variety of inclusion complexes when the size and shape complementarity of host and guest are compatible. Further, interaction of guest molecules with the outer surface of alpha-CD has also been observed. Primarily it is the electrostatic interactions that essentially constitute a driving force for the formation of inclusion complexes. To gain insights for these interactions, the electronic structure and the molecular electrostatic potentials in alpha-, beta-, and gamma-CDs are derived using the hybrid density functional theory employing the three-parameter exchange correlation functional due to Becke, Lee, Yang, and Parr (B3LYP). The present work demonstrates how the topography of the molecular electrostatic potential (MESP) provides a measure of the cavity dimensions and understanding of the hydrogen-bonded interactions involving primary and secondary hydroxyl groups. In alpha-CD, hydrogen-bonded interactions between primary -OH groups engender a "cone-like" structure, while in beta- or gamma-CD the interactions from the primary -OH with ether oxygen in glucose ring facilitates a "barrel-like" structure. Further, the strength of hydrogen-bonded interactions of primary -OH groups follows the rank order alpha CD > beta-CD > gamma-CD, while the secondary hydrogen-bonded interactions exhibit a reverse trend. Thus weak hydrogen-bonded interactions prevalent in gamma-CD manifest in shallow MESP minima near hydroxyl oxygens compared to those in alpha- or beta-CD. Furthermore, electrostatic potential topography reveals that the guest molecule tends to penetrate inside the cavity forming the inclusion complex in beta- or gamma-CD. PMID- 17134169 TI - A coupled Car-Parrinello molecular dynamics and EXAFS data analysis investigation of aqueous Co(2+). AB - We have studied the microscopic solvation structure of Co(2+) in liquid water by means of density functional theory (DFT)-based Car-Parrinello molecular dynamics (CPMD) simulations and extended X-ray absorption fine structure (EXAFS) data analysis. The effect of the number of explicit water molecules in the simulation box on the first and second hydration shell structures has been considered. Classical molecular dynamics simulations, using an effective two-body potential for Co(2+)-water interactions, were also performed to show box size effects in a larger range. We have found that the number of explicit solvent molecules has a marginal role on the first solvation shell structural parameters, whereas larger boxes may be necessary to provide a better description of the second solvation shell. Car-Parrinello simulations were determined to provide a reliable description of structural and dynamical properties of Co(2+) in liquid water. In particular, they seem to describe both the first and second hydration shells correctly. The EXAFS signal was reconstructed from Car-Parrinello simulations. Good agreement between the theoretical and experimental signals was observed, thus strengthening the microscopic picture of the Co(2+) solvation properties obtained using first-principle simulations. PMID- 17134170 TI - DFT/TDDFT study of lanthanide(III) mono- and bisporphyrin complexes. AB - The electronic structure, molecular structure, and electronic spectra of lanthanide(III) mono- and bisporphyrin complexes are investigated using a DFT/TDDFT method. These complexes include YbP(acac), YbP(2), [YbP(2)](+), YbHP(2), and [YbP(2)](-) (where P = porphine and acac = acetylacetonate). To shed some light on the origin of the out-of-plane displacement of Yb in YbP(acac), unligated model systems, namely, planar D(4h) and distorted C(4nu) YbP, were calculated. For comparison, the calculations were also extended to include the C and [Ce(IV)P(2) ](+) systems. Even without an axial ligand, the lanthanide atom lies considerably above the porphyrin plane; the distortion of the YbP molecular structure from a planar D(4h) to the nonplanar C(4nu) symmetry leads to a considerable energy lowering. The axial ligand makes the metal out-of-plane displacement even larger, and it also changes the redox properties of the lanthanide monoporphyrin. The ground-state configurations of YbP(2) and YbHP(2) were determined by considering several possible low-lying states. YbP(2) is confirmed to be a single-hole radical. The special redox properties of the bisporphyrin complexes can well be accounted for by the calculated ionization potentials and electron affinities. The TDDFT results provide a clear description of the UV-vis and near-IR absorption spectra of the various lanthanide porphyrins. PMID- 17134171 TI - The nature of variations of ammonia proton affinity in an argon environment. AB - The presence of solvent molecules, even inert, may significantly influence processes taking place in the gas phase. The reason for the solvent activity may be found in studies of complexes originating from microsolvation. The structure, thermodynamics, and vibrational properties of NH(4)(+)Ar(n) (n = 0-5) and NH(3)Ar(n) (n = 0-4) complexes are presented in this work with the aim to elucidate the effect of microsolvation on protonation/deprotonation processes. The relation between the nature of interactions in cationic and neutral clusters and the proton affinity is studied as a function of the number of ligands in complexes. PMID- 17134172 TI - Asymptotic extrapolation scheme for large-scale calculations with hybrid coupled cluster and molecular dynamics simulations. AB - In this paper we discuss a simple extrapolation scheme based on the asymptotic behavior of the electronic energies considered as functions of cutoff factor for orbital energies corresponding to virtual orbitals. The performance of this approach is illustrated in the context of large-scale dynamic simulations for excitation energies of the cytosine molecule in its native DNA environment. We demonstrate that the extrapolation errors are significantly smaller than the excitation-energy fluctuations, due to the fluctuating environment. PMID- 17134173 TI - Interpretation of the gas-phase solvent deuterium kinetic isotope effects in the S(N)2 reaction mechanism: Comparison of theoretical and experimental results in the reaction of microsolvated fluoride ions with methyl halides. AB - We carried out a comprehensive ab initio calculation and transition-state theory analysis of the solvent and secondary deuterium kinetic isotope effects in the SN2 reactions of microsolvated fluoride ions with methyl halides. Water, methanol, and hydrogen fluoride were used as solvents, and the results are compared with recent experiments. Kinetic isotope effects were dissected into contributions from translations, rotations, and different vibration modes, and the validity of such analysis is also discussed. Excellent agreement was found for some reactions, whereas the agreement was poor for other reactions. We showed that the deviation between theory and experiments is related to the reaction kinetics; a faster reaction produced a kinetic isotope effect that was systematically larger (less inverse) than the calculated value. In addition, we also found that the magnitude of the deviation is proportional to the reaction efficiency. We rationalize the disagreement as a failure of the transition-state theory to model barrierless reactions, and we propose a very simple scheme to interpret these findings and predict the deviation between experimental and theoretical values in those reactions. PMID- 17134175 TI - Nanoscale current modulations in Pr(0.7)Ca(0.3)MnO(3) thin films. AB - Many theoretical and experimental efforts have been focused on the origin of the electric-pulse-induced resistance change effect. However, there are still various reports of the origin supporting either the bulk nature or the interface nature. To resolve the controversies, nanoscale electronic measurements may provide essential clues. In this work, we report microscopic electrical properties of Pr(0.7)Ca(0.3)MnO(3) thin films. The resistance of a single-crystalline grain is not homogeneous in nanometer scale. We deduce that nanoscale inhomogeneity is related to the periodic relaxation of substrate-induced strain, which is caused by the lattice mismatch between the substrate and the thin film. PMID- 17134176 TI - Microscopic inhomogeneity and ultrafast orientational motion in an organic photovoltaic bulk heterojunction thin film studied with 2D IR vibrational spectroscopy. AB - Two-dimensional infrared vibrational spectroscopy is used to examine conformational inhomogeneity and ultrafast orientational motion within local environments of an organic photovoltaic bulk heterojunction thin film. The bulk heterojunction material consists of a mixture of the electron donor poly[2 methoxy-5-(2'-ethylhexyloxy)-1,4-(1-cyanovinylene)phenylene] (CN-MEH-PPV) and the electron acceptor [6,6]-phenyl-C(61)-butyric acid methyl ester (PCBM). PCBM species reside in a distribution of environments within large domains of the molecules that cause their C=O stretch modes to be inhomogeneously broadened. The molecular inhomogeneity also results in frequency dependent vibrational relaxation dynamics. The butyric acid methyl ester group of PCBM undergoes ultrafast wobbling-in-the-cone orientational motion on the 110 fs time scale within a cone semiangle of 29 degrees . The vibrational dynamics are sensitive metrics of molecular order in the material and have implications for charge mobility and degradation phenomena in organic photovoltaic devices. This report represents the first study of organic photovoltaic materials using ultrafast two dimensional infrared vibrational spectroscopy. PMID- 17134177 TI - Is the band gap of pristine TiO(2) narrowed by anion- and cation-doping of titanium dioxide in second-generation photocatalysts? AB - Second-generation TiO(2)-(x)D(x) photocatalysts doped with either anions (N, C, and S mostly) or cations have recently been shown to have their absorption edge red-shifted to lower energies (longer wavelengths), thus enhancing photonic efficiencies of photoassisted surface redox reactions. Some of the studies have proposed that this red-shift is caused by a narrowing of the band gap of pristine TiO(2) (e.g., anatase, E(bg) = 3.2 eV; absorption edge ca. 387 nm), while others have suggested the appearance of intragap localized states of the dopants. By contrast, a recent study by Kuznetsov and Serpone (J. Phys. Chem. B, in press) has proposed that the commonality in all these doped titanias rests with formation of oxygen vacancies and the advent of color centers (e.g., F, F(+), F(++), and Ti(3+)) that absorb the visible light radiation. This article reexamines the various claims and argues that the red-shift of the absorption edge is in fact due to formation of the color centers, and that while band gap narrowing is not an unknown occurrence in semiconductor physics it does necessitate heavy doping of the metal oxide semiconductor, thereby producing materials that may have completely different chemical compositions from that of TiO(2) with totally different band gap electronic structures. PMID- 17134178 TI - Band gap variation in Prussian Blue via cation-induced structural distortion. AB - The charge-transfer band gap of the iron cyanide framework material Prussian Blue and its dependence on the type and location of the charge-compensating interstitial cations (K(+), Rb(+), Cs(+)) are investigated via periodic density functional (DF) calculations. The calculated variation in the band gap magnitude with respect to cation type confirms recent experimental results on cation induced spectral shifts. The role of both the cation interaction with the framework and the cation-induced lattice expansion are examined with respect to their influence on the band gap. The gap magnitude is related to the cation type but is found to be more strongly affected by cation-induced lattice distortion as the cation passes through the material. Our results support the possibility of engineering the electronic structure of Prussian Blue type materials through guest-induced host-framework distortion. PMID- 17134179 TI - Digital logic circuit based on a single molecular system of salicylidene Schiff base. AB - The salicylidene Schiff base N-3,5-dichloro-salicylidene-(S)-alpha phenylethylamine (SPEA) has been synthesized and characterized. Stimulated by one optical input (UV light) and two chemical inputs (OH(-) and Zn(2+)), SPEA undergoes reactions of photochemistry, deprotonation, and complexation. Tailing these reactions by means of the UV-vis absorption spectra and fluorescence spectra, two obvious optical outputs, an absorption band at 323 nm and a fluorescent emission peak at 460 nm, have been obtained. On the basis of encoding binary digits in these inputs and outputs applying positive logic conventions, one monomolecular circuit, which integrates one OR, two NOT, and four AND gates, has been achieved. PMID- 17134180 TI - Controllable synthesis and enhanced electrochemical properties of multifunctional Au(core)Co(3)O(4shell) nanocubes. AB - Multifunctional Au(core)Co(3)O(4shell) nanocubes were synthesized through the introduction of chloroauric acid (HAuCl(4)) into a typical hydrothermal system after a solvothermal process was completed to form metastable Co(3)O(4) hollow nanospheres in the presence of sodium dodecyl benzenesulfonate (SDBS), which served as the surfactant. The strategy suggested that HAuCl(4) played a vital role in the shape transformation and core/shell structure formation, and the sizes of the nanocubes can be tunable through control of the acid concentration. The core/shell structure of the nanocubes was demonstrated by X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and element analysis (EA) measurements. Moreover, Li ion battery measurement indicated that trace Au intercalation altered not only the size and shape of the Co(3)O(4) nanoparticles but also greatly increased their electrochemical properties. These multifunctional nanocubes will be not only helpful to study physical chemistry properties of magnetic nanocrystals but also are expected to find use in many fields such as biomolecular detection and analysis, sensor, electrochemistry, and Li ion batteries. PMID- 17134181 TI - A computational investigation of the different intermediates during organoalkoxysilane hydrolysis. AB - Using a combination of atomistic molecular dynamics (MD) simulations and density functional theory (DFT) calculations, the four steps of hydrolysis of aminopropyl , thiolpropyl-, and butyltrimethoxysilane have been studied. Large box MD simulations at constant pressure and temperature yield appropriate pair distribution functions--which allows us to quantify the number of surrounding water molecules--as well as the density of the systems. These densities serve as input for small box DFT calculations, which allow further geometry optimization and calculation of the electronic structure of the systems. The periodic DFT calculations are compared with gas-phase simulations. In all cases, the first step of hydrolysis is exothermic with the extent depending on the type of silane as well as on the number of hydrogen bonds in the initial stage. PMID- 17134182 TI - Simultaneous phase- and size-controlled synthesis of TiO(2) nanorods via non hydrolytic sol-gel reaction of syringe pump delivered precursors. AB - The simultaneous phase- and size-controlled synthesis of TiO(2) nanorods was achieved via the non-hydrolytic sol-gel reaction of continuously delivered two titanium precursors using two separate syringe pumps. As the injection rate was decreased, the length of the TiO(2) nanorods was increased and their crystalline phase was simultaneously transformed from anatase to rutile. When the reaction was performed by injecting titanium precursors contained in two separate syringes into a hot oleylamine surfactant solution with an injection rate of 30 mL/h, anatase TiO(2) nanorods with dimensions of 6 nm (thickness) x 50 nm (length) were produced. When the injection rate was decreased to 2.5 mL/h, star-shaped rutile TiO(2) nanorods with dimensions of 25 nm x 200 nm and a small fraction of rod shaped anatase TiO(2) nanorods with dimensions of 9 nm x 100 nm were synthesized. Pure star-shaped rutile TiO(2) nanorods with dimensions of 25 nm x 450 nm were synthesized when the injection rate was further decreased to 1.25 mL/h. The simultaneous phase transformation and length elongation of the TiO(2) nanorods were achieved. Under optimized reaction conditions, as much as 3.5 g of TiO(2) nanorods were produced. The TiO(2) nanorods were used to produce dye-sensitized solar cells, and the photoconversion efficiency of the mixture composed of star shaped rutile TiO(2) nanorods and a small fraction of anatase nanorods were comparable to that of Degussa P-25. PMID- 17134183 TI - Nucleating pattern formation in spin-coated polymer blend films with nanoscale surface templates. AB - We use Dip-Pen Nanolithography (DPN) to generate monolayer surface templates for guiding pattern formation in spin-coated polymer blend films. We study template directed pattern formation in blends of polystyrene/poly(2-vinylpyridine) (PS/P2VP) as well as blends of PS and the semiconducting conjugated polymer poly(3-hexylthiophene) (P3HT). We show that acid-terminated monolayers can be used to template pattern formation in PS/P3HT blends, while hydrophobic monolayers can be used to template pattern formation in PS/P2VP blends. In both blends, the polymer patterns comprise laterally-phase separated regions surrounded by vertically separated bilayers. We hypothesize that the observed patterns are formed by template-induced dewetting of the bottom layer of a polymer bilayer during the spin-coating process. We compare the effects of template feature size and spacing on the resulting polymer patterns with predictions from published models of template-directed dewetting in thin films and find the data in good agreement. For both blends we observe that a minimum feature size is required to nucleate dewetting/phase separation. We find this minimum template diameter to be approximately 180 nm in 50/50 PS/P2VP blends, and approximately 100 nm in 50/50 PS/P3HT blends. For larger template diameters, PS/P2VP blends show evidence for pattern formation beginning at the template boundaries, while PS/P3HT blends rupture randomly across the template features. PMID- 17134184 TI - Energy and charge transfer dynamics in fully decorated benzyl ether dendrimers and their disubstituted analogues. AB - We examine the photophysics of a series of molecules consisting of a benzthiadiazole core surrounded by a network of benzyl ether arms terminated by aminopyrene chromophores, which function as both energy and electron donors. Three classes of molecules are studied: dendrimers whose peripheries are fully decorated with aminopyrene donors (F), disubstituted dendrimers whose peripheries contain only two donors (D), and linear analogues in which a pair of benzyl ether arms link two donors to the central core (L). The electronic energy transfer (EET) and charge transfer (CT) rates are determined by fluorescence lifetime measurements on the energy donors and electron acceptors, respectively. In all three types of molecules, the EET time scales as the square root of the generation number G, consistent with the flexible nature of the benzyl ether framework. Transient anisotropy measurements confirm that donor-donor energy hopping does not play a major role in determining the EET times. The CT dynamics occur on the nanosecond time scale and lead to stretched exponential decays, probably due to conformational disorder. Measurements at 100 degrees C confirm that conformational fluctuations play a role in the CT dynamics. The average CT time increases with G in the L and D molecules but decreases for the F dendrimers. This divergent behavior as G increases is attributed to the competing effects of larger donor-acceptor distances (which lengthen the CT time) versus a larger number of donors (which shorten the average CT time). This work illustrates two important points about light-harvesting and charge-separation dendrimers. First, the use of a flexible dendrimer framework can lead to a more favorable scaling of the EET time (and thus the light-harvesting efficiency) with dendrimer size, relative to what would be expected for a fully extended dendrimer. Second, fully decorated dendrimers can compensate for the distance dependent slowdown in CT rate as G increases by providing additional pathways for the CT reaction to occur. PMID- 17134185 TI - Laser ablation condensation, coalescence, and phase change of dense gamma Al(2)O(3) particles. AB - Dense gamma-Al(2)O(3)condensates, with residual stress up to 3 GPa and ranging from nanometer to an unexpected micrometer size, were formed by pulsed laser ablation on Al target under oxygen background gas for a very rapid heating and cooling effect. Analytical electron microscopic observations indicated such nanoparticles tended to coalesce over {111} facets to form multiple twin and tilt boundary. The micrometer-size particles changed, upon electron irradiation, into metastable orthorhombic delta form full of twin variants and faults parallel to {100}. PMID- 17134186 TI - Diphenyloligothiophene-based dichromophoric macrocycles and their ability to form donor/acceptor complexes. AB - A series of four dichromophoric rigid macrocycles 6a-6d, two with diphenyloligothiophene chromophores, the other two with more electron-rich diphenyl-EDOT or diphenyl-bis-EDOT chromophores, have been synthesized. The absorption spectrum of the diphenyl-bis-EDOT based macrocycle 6d displayed the most pronounced vibronic resolution with a well-resolved 0-0 transition, indicating a fully planarized geometry of the diphenyl-bis-EDOT chromophores. The (1)H NMR spectra of the macrocycles displayed weak to moderate chemical shifts of characteristic signals upon addition of pi-conjugated oligonitro-9-fluorenone acceptors. X-ray single-crystal analysis showed that columnar pi-stacked donor/acceptor complexes are formed with the stacks composed of alternating donor and acceptor molecules. The stoichiometry of the crystalline, dark-colored complexes was found to be 1:1 by elemental analysis and integration of the (1)H NMR peaks. The complex formation is accompanied by remarkably large Stern-Volmer constants of fluorescence quenching. PMID- 17134187 TI - Excited state dynamics and nonlinear absorption of a pyrazinoporphyrazine macrocycle carrying externally appended pyridine rings. AB - The multiphoton absorption properties of the tetrakis-2,3-[5,6-di(2 pyridyl)pyrazino]porphyrazinato(monoacquo)-Mg(II) complex [Py(8)TPyzPzMg(H(2)O)] (1) are reported and interpreted. The nonlinear optical behavior of 1 and the characterization of the excited states important for the nonlinear absorption process were studied at the pump frequency of the second harmonic generation of a Nd:YAG laser in the nanosecond time regime. It was found that complex 1 shows a very good optical limiting performance at 532 nm, which derives from two processes: (a) a reverse-saturable absorption process, which involves a triplet excited state at low intensities, and (b) a two-photon absorption process at higher intensities, which is due to the formation of the radical monoanion of 1, [Py(8)TPyzPzMg(H2O)](.-), during the photoreduction of the triplet state. The participation of a monoanion in determining the overall nonlinear absorption behavior of 1 is found, for the first time, for a tetrapyrrolic system. One can deduce that the involvement of the monoanion derives from the electron withdrawing effect of the dipyridinopyrazino fragments externally attached to the porphyrazine core which make the reduced form of 1 easily accessible. These results suggest a modification of tetrapyrrolic systems with new nonlinear absorption properties. PMID- 17134188 TI - Quantum chemical analysis of electronic structure and n- and p-type charge transport in perfluoroarene-modified oligothiophene semiconductors. AB - Density-functional theory (DFT) is employed to investigate the structural, electronic, and transport properties of several isomeric fluoroarene oligothiophene-based semiconductors. Three oligothiophene systems varying in the perfluoroarene group positions within the molecule are studied to understand the electronic structure leading to the observed mobility values and to the n- or p type behavior in these structures. Analyses of both intermolecular interactions in dimers and extended interactions in crystalline structures afford considerable insight into the electronic properties and carrier mobilities of these materials, as well as the polarity of the charge carriers. From the calculated carrier effective masses, we find that sterically governed molecular planarity plays a crucial role in the transport properties of these semiconductors. Our calculations correlate well with experimentally obtained geometries, highest occupied molecular orbital (HOMO)/lowest-unoccupied molecular orbital (LUMO) energies, and the experimental carrier mobility trends among the systems investigated. PMID- 17134189 TI - Determination of carbon nanotube density by gradient sedimentation. AB - Density gradient centrifugation is a high-resolution technique for the separation and characterization of large molecules and stable complexes. We have analyzed various nanotube structures by preparative centrifugation in sodium metatungstate water solutions. Bundled, isolated and acid-treated single-walled nanotubes (SWNTs) and multiwall nanotubes (MWNTs) formed sharp bands at well-defined densities. The structure of the material in each band was confirmed by transmission electron microscopy and Raman spectroscopy. Our data suggest respective densities of 1.87, 2.13, 1.74, and 2.1 g/cm(3) for bundled, isolated, and acid-treated SWNTs and MWNTs. These measured results compare well with their calculated densities. PMID- 17134190 TI - Local and network structure of thermoreversible polyrotaxane hydrogels based on poly(ethylene glycol) and methylated alpha-cyclodextrins. AB - Thermoreversible gelation and microphase formation of aqueous solutions of a methylated polyrotaxane (MePR) were investigated by means of differential scanning microcalorimetry, rheometry, and X-ray diffractometry (XRD). The aqueous solutions of MePR show a lower critical solution temperature (LCST) and form an elastic gel with increasing temperature. The sol-gel transition of the MePR solutions was induced by formation and deformation of aggregates of methylated alpha-cyclodextrins (alpha-CDs) of polyrotaxane due to hydrophobic dehydration and hydration, respectively. The XRD investigation revealed localization and highly ordered arrangement of methylated alpha-CDs along the PEG chain in the gel. The arrangement of CDs was also reflected by the changes in elasticity and long relaxation behavior of the solution around the sol-gel transition. The quasiequilibrium shear modulus of MePR solutions showed the critical phenomena against temperature. The scaling exponents measured at two different concentrations were almost equal to the values predicted by a gel percolation theory. Therefore, the heat-induced gelation of aqueous MePR solutions is well explained by a model in which clusters assembled with methylated alpha-CDs are gradually connected to the network as the temperature increases. PMID- 17134191 TI - Effect of substrate surface on dewetting behavior and chain orientation of semicrystalline block copolymer thin films. AB - Three symmetrical semicrystalline oxyethylene/oxybutylene block copolymers (EmBn) were spin-coated on different substrates including silicon, hydrophobically modified silicon, and mica. The effects of surface property on the dewetting behavior of EmBn thin films and the chain orientation of the crystalline block were investigated with atomic force microscopy and grazing incidence X-ray diffraction . The EmBn thin films on silicon exhibit an autophobic dewetting behavior, while ordinary dewetting occurs for the thin films on modified silicon. It was observed that the stems of the E crystals in the first half-polymer layer contacting the mica surface were parallel to the surface, in contrast to the perpendicular chain orientation of the other polymer layers and of the first half polymer layer on silicon. This is attributed to the strong interaction between the E block and mica, verified by infrared spectra. PMID- 17134192 TI - Photoisomerization of azobenzene derivatives in nanostructured silica. AB - A series of derivatized azobenzene molecules are synthesized such that one of the phenyl groups can be chemically bonded to mesostructured silica and the other, derivatized with dendrons, is free to undergo large-amplitude light-driven motion. The silica frameworks on which the motion takes place are either 150 nm thick films containing ordered hexagonal arrays of tubes (inner diameter about 2 nm) containing the bonded azobenzenes, or particles (about 500 nm in diameter) containing the same ordered arrays of functionalized tubes. The photoisomerization yields and the rate constants for the thermal cis to trans back-reaction of the azobenzenes in the tubes are measured and compared to those of the molecules in solution. The rate constants decrease with increasing size of the dendrons. Fluorescence spectra of the cis and trans isomers in the pores show that the photoisomerization in the nanostructured materials is selectively driven by specific wavelengths of light and is reversible. PMID- 17134193 TI - MnS hierarchical hollow spheres with novel shell structure. AB - High yields of MnS microspheres with novel hierarchical structure were prepared through a simple solution method. Field emission scanning electron microscopy and transmission electron microscopy analyses reveal that the microsphere has a core shell structure: the interior hollow sphere is covered by a shell consisting of nanorod arrays. Interestingly, the nanorod is a wurtzite (WZ)/zinc blende (ZB) phase admixture with a large amount of stacking faults/twins. The alternation of WZ and ZB along the growth direction of the nanorod enables it to exhibit the features of a quantum well. Furthermore, the WZ/ZB admixture structure could also be regarded as a type II homomaterial heterostructure. All these features imply that the novel core-shell structure has great potential for applications, among them the quantum well photoelectrical and heterostructure photoconduction fields. PMID- 17134194 TI - Aggregated CdS quantum dots: Host of biomolecular ligands. AB - In this contribution, we have studied structural and photophysical properties of aggregated CdS quantum dots (QDs) capped with 2-mercaptoethanol in aqueous medium. The hydrodynamic diameter of the nanostructures in aqueous solution was found to be approximately 160 nm with the dynamic light scattering (DLS) technique, which is in close agreement with atomic force microscopy (AFM) studies (diameter approximately 150 nm). However, the UV-vis absorption spectroscopy, powder X-ray diffraction (XRD), and transmission electron microscopy (TEM) studies confirm the average particle size (QD) in the nanoaggregate to be 4.0 +/- 0.5 nm. The steady-state and time-resolved photoluminescence studies on the QDs further confirm preservation of electronic band structure of the QDs in the nanoaggregate. To study the nature of the nanoaggregate we have used small fluorescent probes, which are widely used as biomolecular ligands (2,6-p toluidinonaphthalene sulfonate (TNS) and Oxazine 1), and found the pores of the aggregate to be hydrophobic in nature. The significantly large spectral overlap of the host quantum dots (donor) with that of the guest fluorescent probe Oxazine 1 (acceptor) allows us to carry out Forster resonance energy transfer (FRET) studies to estimate average donor-acceptor distance in the nanostructure, found to be approximately 25 Angstrom. The quantum dot aggregate and the characterization techniques reported here could have implications in the future application of the QD-nanoaggregate as host of small ligand molecules of biological interest. PMID- 17134195 TI - Alcohol and proton transport in perfluorinated ionomer membranes for fuel cells. AB - To clarify the transport mechanisms of alcohols and proton in perfluorosulfonated ionomer (PFSI) membranes for fuel cells, four membranes having different equivalent weight (EW) values were examined. Membranes were immersed in methanol, ethanol, and 2-propanol to prepare a total of 12 samples, and membrane swelling, mass (alcohol and proton) transports, and interactions between alcohols and proton were investigated systematically in the fully penetrated state. The membrane expansion fraction theta and alcohol content lambda increased with decreasing the EW value for all the samples. The self-diffusion coefficients (D's) of the alkyl group and of OH (including protons) were measured separately by the pulsed-gradient spin-echo (PGSE)-NMR method and the D's also increased with decreasing the EW value. These results implied that the alcohols penetrate into the hydrophilic regions of the PFSI membranes and diffuse through the space expanded by the alcohols. The ionic cluster regions formed by the alcohols resemble those induced by water in the water swollen membrane, where protons dissociated from sulfonic acid groups transport through the regions together with water molecules. The D values decreased with increasing the molecular weight of alcohols. This trend was supported by activation energies Ea estimated from the Arrhenius plots of D in the temperature range from 30 to -40 degrees C. The PGSE NMR measurements also revealed that protons move faster than the alkyl groups in the membranes. The proton transport by the Grotthuss (hopping) mechanism was facilitated by the increase of the alcohol content and the decrease of the molecular weight. This result was also supported by the experimental results of proton conductivity kappa and mobility u(H(+)). Density functional theory (DFT) calculations of the interaction energy DeltaE(int) between proton and alcohol (including OH) showed that the /DeltaE(int)/ increases with increasing the molecular weight of alcohols, which is in a inverse relationship with the kappa and u(H(+)) values. The proton transport depends strongly on the DeltaE(int) in the membranes. PMID- 17134196 TI - Synthesis and characterization of highly magnetized nanocrystalline Co(30)Fe(70) alloy by chemical reduction. AB - Co(30)Fe(70) nanoparticles with mean particle size of about 8 nm were successfully synthesized by the chemical reduction of cobalt chloride and iron chloride with borohydride as a reducing agent in aqueous solution. The composition and size of the Co(30)Fe(70) nanoparticles were optimized by controlling the molar ratio of starting materials, reaction time, and dropping rate of aqueous reducing agent. As alloy powders prepared by chemical reduction tend to be amorphous in the as-synthesized state, the as-precipitated Co(30)Fe(70) nanoparticles were heat-treated to achieve crystallinity at the different temperatures for 1 h. The Co(30)Fe(70) nanocrystallite by chemical reduction shows excellent soft magnetic behavior, such as high permeability, negligible coercivity, and high saturation magnetization like that of Co(30)Fe(70) bulk. PMID- 17134197 TI - Hybrid junctions of zinc(II) and magnesium(II) phthalocyanine with wide-band-gap semiconductor nano-oxides: spectroscopic and photoelectrochemical characterization. AB - The optical properties of zinc phthalocyanine (ZnIIPc) and magnesium phthalocyanine (MgIIPc) in DMSO and DMF solutions have been extensively investigated, and the photoelectrochemical behaviors of layer-by-layer hybrid junctions formed of the two metallo(II) phthalocyanines (MIIPcs) and wide-band gap colloidal semiconductors, namely, ZnO and TiO2 nanocrystals (NCs), have been probed. Different experimental conditions, such as the Pc center metal ion, dye concentration, and solvent identity, were investigated in order to elucidate their effects on the photoelectrochemical performances of the prepared heterojunctions. Finally, thermal treatment of either dye and NC films and control of the NC shape and surface chemistry were also studied and, interestingly, were found to be critical in affecting the performance of photochemical sensitization processes, occurring at the dye/oxide and oxide/solution interfaces. PMID- 17134198 TI - Accurate sizing of nanoparticles using confocal correlation spectroscopy. AB - The ability to accurately size low concentrations of nanoscale particles in small volumes is useful for a broad range of disciplines. Here, we characterize confocal correlation spectroscopy (CCS), which is capable of measuring the sizes of both fluorescent and nonfluorescent particles, such as quantum dots, gold colloids, latex spheres, and fluorescent beads. We accurately measured particles ranging in diameter from 11 to 300 nm, a size range that had been difficult to probe, owing to a phenomenon coined biased diffusion that causes diffusion times, or particle size, to deviate as a function of laser power. At low powers, artifacts mimicking biased diffusion are caused by saturation of the detector, which is especially problematic when probing highly fluorescent or highly scattering nanoparticles. However, at higher powers (>1 mW), autocorrelation curves in both resonant and nonresonant conditions show a structure indicative of an increased contribution from longer correlation times coupled with a decrease in shorter correlation times. We propose that this change in the autocorrelation curve is due to the partial trapping of the particles as they transit the probe volume. Furthermore, we found only a slight difference in the effect of biased diffusion when comparing resonant and nonresonant conditions. Simulations suggest the depth of trapping potential necessary for biased diffusion is > 1 k(B)T. Overcoming artifacts from detector saturation and biased diffusion, CCS is particularly advantageous due to its ability to size particles in the small volumes characteristic of microfluidic channels and aqueous microdroplets. We believe the method will find increasing use in a wide range of applications in measuring nanoparticles and macromolecular systems. PMID- 17134199 TI - Highly ordered superlattices from polydisperse Ag nanoparticles: a comparative study of fractionation and self-correction. AB - We have examined two different routes to construct highly ordered two- or three dimensional (2D or 3D) superlattice structures from hydrophilic polydisperse mercaptosuccinic acid (MSA)-modified Ag nanoparticles of the average size of 2.5 nm. First, polydisperse particles were fractionized by the polyacrylamide gel electrophoresis (PAGE) method. Due to the size-dependent migration under the electric field, the particles were isolated into a series of gel bands and each band contained particles with significantly narrow size distribution. Subsequent to phase transfer into chloroform by cationic surfactant, long-range 2D superlattices were simultaneously formed on the substrate upon evaporation of chloroform. Second, 3D superlattices were directly grown at an air-water interface from the polydisperse bulk dispersion by diffusion of HCl vapor without any pretreatment for the size narrowing. The influence of diffusion rate of HCl was also studied. The achievement of 3D superlattices via this route was ascribed as a long-time self-correction process. Furthermore, it was revealed that the superlattice structures obtained by the above two procedures exhibited distinct features though the starting material was the same MSA-Ag nanoparticles. The surface distance of core between component particles, the orientation of particles inside the superlattice, and the process of superlattice formation were comprehensively studied. We confirmed that each growth process depended on a corresponding self-assembly mechanism. PMID- 17134200 TI - Controlled synthesis, characterization, and catalytic properties of Mn(2)O(3) and Mn(3)O(4) nanoparticles supported on mesoporous silica SBA-15. AB - A method established in the present study has proven to be effective in the synthesis of Mn(2)O(3) nanocrystals by the thermolysis of manganese(III) acetyl acetonate ([CH(3)COCH=C(O)CH(3)](3)-Mn) and Mn(3)O(4) nanocrystals by the thermolysis of manganese(II) acetyl acetonate ([CH(3)COCH=C(O)-CH(3)](2)Mn) on a mesoporous silica, SBA-15. In particular, Mn(2)O(3) nanocrystals are the first to be reported to be synthesized on SBA-15. The structure, texture, and electronic properties of nanocomposites were studied using various characterization techniques such as N2 physisorption, X-ray diffraction (XRD), laser Raman spectroscopy (LRS), temperature-programmed reduction (TPR), transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS). The results of powder XRD at low angles show that the framework of SBA-15 remains unaffected after generation of the manganese oxide (MnO(x)) nanoparticles, whereas the pore volume and the surface area of SBA-15 dramatically decreased as indicated by N2 adsorption-desorption. TEM images reveal that the pores of SBA-15 are progressively blocked with MnO(x) nanoparticles. The formation of the hausmannite Mn(3)O(4) and bixbyite Mn(2)O(3) structures was clearly confirmed by XRD. The surface structures of MnO(x) were also determined by LRS, XPS, and TPR. The crystalline phases of MnO(x) were identified by LRS with corresponding out-of plane bending and symmetric stretching vibrations of bridging oxygen species (M-O M) of both MnO(x) nanoparticles and bulk MnO(x). We also observed the terminal Mn=O bonds corresponding to vibrations at 940 and 974 cm-1 for Mn(3)O(4)/SBA-15 and Mn(2)O(3)/SBA-15, respectively. These results show that the MnO(x) species to be highly dispersed inside the channels of SBA-15. The nanostructure of the particles was further identified by the TPR profiles. Furthermore, the chemical states of the surface manganese (Mn) determined by XPS agreed well with the findings of LRS and XRD. These results suggest that the method developed in the present study resulted in the production of MnO(x) nanoparticles on mesoporous silica SBA-15 by controlling the crystalline phases precisely. The thus-prepared nanocomposites of MnO(x) showed significant catalytic activity toward CO oxidation below 523 K. In particular, the MnO(x) prepared from manganese acetyl acetonate showed a higher catalytic reactivity than that prepared from Mn(NO(3))2. PMID- 17134201 TI - Conductometric study of precursor compound formation during wet-chemical synthesis of nanocrystalline hydroxyapatite. AB - Conductometry was employed to study the phase evolution of calcium phosphate compounds during the wet-chemical synthesis of hydroxyapatite (HA). Calcium hydroxide and orthophosphoric acid were used to prepare HA at various temperatures ranging from 30 to 95 degrees C. The electrical conductivity and pH of the reaction mixture were measured at regular intervals of time during acid addition, and the rate of change of conductivity was used to decipher the end point of the reaction. Our previous studies have shown that the end product of this reaction route yields mildly carbonated crystalline HA. The trend of the change in conductivity with time was similar at all temperatures. The conductivity curves were divided into three regions based on the variation in slope of the curves. The slope of the curves decreased with increasing temperatures in the first two regions, and the slope is greater in the second region than in the first. From the conductivity and pH measurement results, the possible precursor phase was identified and it had the composition Ca(3)(PO(4))(2). The kinetics of phase transformation was also analyzed and compared to previous work. The similarities between this work and the traditionally adapted experimental work for phase formation and transformation kinetics are highlighted, and the novelty in the current work is discussed. PMID- 17134202 TI - Crystallization behaviors of poly(3-hydroxybutyrate) and poly(l-lactic acid) in their immiscible and miscible blends. AB - By adjusting the molecular weight of the poly(l-lactic acid) (PLLA) component in poly(3-hydroxybutyrate) (PHB)/PLLA blends, we investigated the crystallization behaviors of the two components in their immiscible and miscible 50:50 blends by real time infrared (IR) spectroscopy. In the immiscible PHB/PLLA blend, the stepwise crystallization of PHB and PLLA was realized at different crystallization temperatures. PLLA crystallizes first at a higher temperature (120 degrees C). Its crystallization mechanism from the immiscible PHB/PLLA melt is not affected by the presence of the PHB component, while its crystallization rate is substantially depressed. Subsequently, in the presence of crystallized PLLA, the isothermal melt-crystallization of PHB takes place at a lower temperature (90 degrees C). It is interesting to find that there are two growth stages for PHB. At the early stage of the growth period, the Avrami exponent is 5.0, which is unusually high, while in the late stage, it is 2.5, which is very close to the reported value (n approximately 2.5) for the neat PHB system. In contrast to the stepwise crystallization of PHB and PLLA in the immiscible blends, the almost simultaneous crystallization of PHB and PLLA in the miscible 50:50 blend was observed at the same crystallization temperature (110 degrees C). Detailed dynamic analysis by IR spectroscopy has disclosed that, even in such apparently simultaneous crystallization, the crystallization of PLLA actually occurs faster than that of PHB. It has been found that, both in the immiscible and miscible blends, the crystallization dynamics of PHB are heavily affected by the presence of crystallized PLLA. PMID- 17134203 TI - Molybdenum trioxide nanostructures: the evolution from helical nanosheets to crosslike nanoflowers to nanobelts. AB - MoO(3) nanostructures with different morphologies, such as helical nanosheets, crosslike nanoflowers, and nanobelts, have been synthesized on a large scale by an environmentally friendly chemical route. The evolution process from helical nanosheets to crosslike nanoflowers to nanobelts is observed for the first time. The influences of reaction time and the molar ratio of molybdenum and H(2)O(2) on the morphologies of MoO(3) nanostructures have been investigated. The synthetic process is environmentally friendly and may be extended to synthesize nanostructures of other metal (W, Ti, and Cr) oxides. PMID- 17134204 TI - Quenching of IR luminescence of erbium, neodymium, and ytterbium beta-diketonate complexes by ligand C-H and C-D bonds. AB - The effect of CH and CD quenching on the luminescence lifetime of Er(3+) Nd(3+) and Yb(3+) in the Cs[Ln(HFA)(4)] system has been quantified, and we have shown that for Er(3+) ions the quenching is dominated by the nearest neighbor CH oscillators, whereas for Nd(3+) ions the roles of more distant CH oscillators and nearest neighbor CD oscillators are important. PMID- 17134205 TI - Enhanced intra-aggregate charge separation from binary excitons in mixed J aggregates of cyanine dyes. AB - Well-ordered, two-dimensional, amalgamation-type J-aggregates consisting of two different kinds of structurally analogous anionic cyanine dyes were self assembled on an amino-functionalized Au(111) electrode surface. Similar binary J aggregates, though considerably less ordered in either one- or three-dimensional structure, could also be grown easily in a concentrated ( approximately 1.5 M) aqueous solution of NaCl. These binary J-aggregates are characterized by a singly peaked J-band associated with a unique molecular exciton shared by the two different dye components coherently. This cooperative binary excitonic state can live with substantial (more than 0.1 eV) energy gaps between the HOMO (highest occupied molecular orbital) and LUMO (lowest unoccupied molecular orbital) levels of the two dyes mixed together. These energy gaps allow the binary exciton to relax very rapidly to an intra-aggregate charge-separated state in good competition with the metal-induced quenching of the excited-state energy. The enhanced charge separation leads to a high-efficiency photocurrent generation with a simple binary J-aggregate-coated Au(111) electrode in a reversible redox solution and results in strong quenching of the fluorescence from the corresponding aggregate grown in solution free from substrate-induced quenching. PMID- 17134206 TI - MnO octahedral nanocrystals and MnO@C core-shell composites: synthesis, characterization, and electrocatalytic properties. AB - We present a simple and facile synthesis of MnO octahedral nanocrystals and MnO@C core-shell composite nanoparticles. The synthesis is accomplished by a single step direct pyrolysis of cetyltrimethylammonium permanganate in specially made Let-lock union cells. The products are characterized by HRSEM, HRTEM, Raman spectroscopy, and cyclic voltammetry (CV). The product consists mainly of octahedral MnO nanocrystals and MnO coated with carbon (MnO@C). The core-shell particles are observed only when the core size is smaller than 150 nm. The shape of the nanocrystals can be controlled by varying parameters such as reaction temperature and duration. As the temperature increases from 600 to 800 degrees C, the octahedral MnO crystals observed are without any carbon shell. The effect of time and temperature on the octahedral MnO nanocrystal formation is described. The electrocatalytic activities of the products are studied for oxygen reduction reaction in aqueous basic medium and are compared with bulk MnO. The MnO nanocrystals and core-shell composites exhibit higher activity than that of bulk MnO. PMID- 17134207 TI - EPR and DFT study on the stabilization of radiation-generated methyl radicals in dehydrated Na-A zeolite. AB - Electron paramagnetic resonance (EPR) spectroscopy was applied to study paramagnetic species stabilized in Na-A zeolite exposed to gaseous methane and gamma-irradiated at 77 K. Two types of EPR spectra were recorded during thermal annealing of zeolite up to room temperature. Owing to the results for the zeolite exposed to (13)CH(4) the multiplet observed at 110 K was assigned to a (. )CH(3)...Na(+) complex. After decay of the multiplet, the isotropic quartet of methyl radical was recorded in the temperature range of 170-280 K. On the basis of the EPR parameters it is postulated that (.-)CH(3) radicals in this temperature region are able to freely rotate inside the zeolite cage. The structures of the (.-)CH(3)...Na(+) adsorption complex and respective hyperfine coupling constants were calculated by applying DFT quantum chemical methods. Two different models were applied to represent the zeolite framework: the 6T structure of one six-membered ring and the 3T cluster. The hyperfine coupling constants calculated for the (.-)CH(3)...Na(+) adsorption complex for both applied models show very good agreement with those obtained experimentally. PMID- 17134208 TI - A thermodynamic model for the shape and stability of twinned nanostructures. AB - Although thermodynamically metastable, planar defects are often observed in many faceted nanomaterials including nanocrystals, nanorods, and nanowires, even after annealing. These planar defects include contact twins and (intrinsic or extrinsic) stacking faults, and are usually neglected by most analytical models. For example, many bulk metals have the face-centered cubic structure, but small nanocrystals and nanorods of the same material often exhibit various structural and morphological modifications such as single or multiple symmetric twinning, as well as 5-fold cyclic twinning resulting in decahedral and truncated decahedral nanostructures. Presented here is a general analytical model for the investigation of nanomaterials of arbitrary shape, and with any configuration of planar defects. The model is tested for the case of twinning in unsupported gold nanocrystals and nanorods, and is shown to give results in excellent agreement with experimental and computational studies reported in the literature. PMID- 17134209 TI - Conduction mechanism of Aviram-Ratner rectifiers with single pyridine-sigma-C(60) oligomers. AB - We present the electron transport of pyridyl aza[60]fulleroid oligomers, abbreviated as C(60)NPy, which is based on the donor-barrier-acceptor (D-sigma-A) architecture, at a single molecular scale using scanning tunneling microscopy. A rectifying effect is observed in the current-voltage characteristics. The theoretical calculation shows that the highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) are well localized either on the Py moiety (donor) or on the C(60) moiety (acceptor), indicating the sigma bridge decouples the LUMO and the HOMO of the donor and the acceptor, respectively. This structure accords well with the unimolecular rectifying model proposed by Aviram and Ratner [Chem. Phys. Lett. 1974, 29, 277]. The mechanism of the rectifying effect is understood by analyzing in detail the electron transport through energy levels of the donor and the acceptor of the C(60)NPy molecules. By directly comparing the experimental conductance peaks and the calculated density of states of the C(60)NPy, we find that the observed rectification is attributed to the asymmetric positioning of the LUMOs and the HOMOs of both sides of the acceptor and the donor of the C(60)NPy molecules with respect to the Fermi level of the electrodes. When a main voltage drop is over the molecule-electrode vacuum junction but a small fraction over the molecule itself, the shift of the energy levels between the donor and the acceptor will be small. This behavior deviates from the original proposal by Aviram and Ratner in which a large shift of the energy level is expected. PMID- 17134210 TI - Catalytically driven colloidal patterning and transport. AB - We recently reported the convection and pattern formation of tracers caused by a catalytically generated electric field. The electric field arises due to the heterogeneous electrochemical reduction and oxidation of hydrogen peroxide (H2O2) on silver (Ag) and gold (Au), respectively.1 Here we describe an electrokinetic model, developed in conjunction with experiments, that explains the details of the convection and pattern formation phenomenon. The model also enables the measurement of reaction kinetic parameters that are otherwise difficult to obtain. This quantitative model serves as a platform for the modeling of other catalytic redox systems and systems with broken symmetries. PMID- 17134211 TI - Structure, wrinkling, and reversibility of Langmuir monolayers of gold nanoparticles. AB - The assembly of nanoparticles into large, two-dimensional structures provides a route for the exploration of collective phenomena among mesoscopic building blocks. We characterize the structure of Langmuir monolayers of dodecanethiol ligated gold nanoparticles with in situ optical microscopy and X-ray scattering. The interparticle spacing increases with thiol concentration and does not depend on surface pressure. The correlation lengths of the Langmuir monolayer crystalline domains are on the order of five to six particle diameters. Further compression of the monolayers causes wrinkling; however, we find that wrinkled monolayers with excess thiol can relax to an unwrinkled state following a reduction of surface pressure. A theoretical model based on van der Waals attraction and tunable steric repulsion is adopted to explain this reversibility. PMID- 17134212 TI - Polymer-surfactant layered heterostructures by electropolymerization of phenosafranine in Langmuir-Blodgett films. AB - Langmuir-Blodgett (LB) films of the water-soluble dye phenosafranine (PS) have been prepared by its adsorption from aqueous dye solution to an arachidic acid (AA) monolayer at the air-water interface. Atomic force microscopy (AFM) images of the LB films revealed the effect of change in pH of deposition on the degree of complexation of AA with the PS dye. Well-defined circular islands and holes were observed which disappeared with the increase in pH. Polarized absorption studies indicated that the dye molecules are oriented uniaxially with their long axis titled at a constant angle to the surface normal of the LB film. Within the restricted geometry of the LB film, the PS dye was electropolymerized to form a two-dimensional film of poly(phenosafranine) sandwiched between arachidic acid layers. The film was characterized by IR spectroscopy, cyclic voltammetry, and AFM. X-ray diffraction studies reveal the presence of a layer structure in the AA PS LB film before and after polymerization. The polymer film showed highly anisotropic electrical conductivity of ca. 10 orders of magnitude. This indicates the formation of two-dimensional polyPS layers between arachidic acid layers resulting in a layered heterostructure film having alternate conducting and insulating regions. Also, the conductivity of the polyPS prepared from LB film was found to be approximately 2.5 times higher than the conductivity of polyPS prepared by solution polymerization method. PMID- 17134213 TI - Theoretical study of CCl(4) adsorption and hydrogenation on a Pt (111) surface. AB - The adsorption and hydrogenation of carbon tetrachloride (CCl(4)) on a Pt (111) surface have been investigated using density functional theory (DFT). We have performed calculations on the adsorption energies and structures of CCl(4) on four different adsorption sites of a Pt (111) surface using the full adsorbate geometry optimization method. The results show that the adsorption energy of all of the potential sites is less than -17 kcal/mol, which indicates that CCl(4) is physiosorbed on a Pt (111) surface through van der Waals interactions. The dissociation and hydrogenation pathways were investigated by a transition state search. For the Pt(15), Pt(19), and Pt(25) cluster surfaces, the activation energies of dissociation obtained in this work are 15.69, 16.94, and 16.77 kcal/mol, respectively. The hydrogenation of CCl(3). was studied at the on-top site of the Pt(15) cluster, and the calculated activation energy is 5.06 kcal/mol. The small activation energies indicate that the Pt (111) surface has high catalytic activity for the CCl(4) hydrogenation reaction. In addition, the Hirshfeld population analysis reveals that the charge transfer from the Pt (111) surface to the adsorbates occurs in both the dissociation and hydrogenation pathways. PMID- 17134214 TI - Solubility of hydrogen in PdAg and PdAu binary alloys using density functional theory. AB - The present work deals with the study of palladium-silver (PdAg) and palladium gold (PdAu) binary alloys over a broad range of temperatures and alloy compositions using density functional theory (DFT) to find possible conditions where the solubility of hydrogen (H) is significantly higher than that of pure palladium (Pd). Several alloy structures, such as Pd(100-x)Ag(x) with x = 14.81, 25.93, 37.04, and 48.51, Pd(100-x)Aux with x = 14.81, 25.93, and 37.04, and Pd(100-x)Cu(x) with x = 25.93 and 48.51 were considered. The lattice constants of these structures were optimized using DFT, and relaxed structures were used for the estimation of binding energy. It was found that the solubility of H in PdAg is higher than pure Pd with a maximum at approximately 30% Ag at 456 K. Also, the solubility of PdAu alloys was higher than pure Pd with a maximum at about 20% Au with a solubility 12 times higher than that of pure Pd. It was found that for a 3.7% H concentration in a PdAg alloy, a cell expansion of 0.15-0.2% occurs, which if ignored may affect the individual binding energy of the O-site by approximately 3.56% and may affect the predicted solubility by approximately 11.8%. PMID- 17134215 TI - Water adsorption and diffusion on NaCl(100). AB - At low coverage and temperature the water-surface interaction determines the adsorption geometry of the water molecule on the NaCl(100) surface. However, at room temperature the molecules are also able to move on the surface and form islands where the water molecules are held together by hydrogen bonds. As a step toward the description of such complex phenomenology, in this work we have used density functional theory calculations to study the most favorable adsorption geometry of an isolated water molecule and the energy barriers associated with different hopping mechanisms between equivalent adsorption configurations on this surface. We propose different hopping processes that can be classified as translations, if the molecule moves from one adsorption site to the adjacent one, or reorientations, if the molecule only changes its orientation on the surface and remains in the same adsorption site. The straightforward parallel translation of the water molecule along the surface exhibits the highest barrier. All other processes, either translations or reorientations, involve the rotation of the water molecule around certain axes and present much smaller barriers (at least 50% smaller). To obtain a net movement of the molecule along the surface it is always necessary to combine one of these translational and reorientational processes. Such combinations provide favorable and plausible pathways for the diffusion of the water molecule on the NaCl(100) substrate. PMID- 17134216 TI - Metal acetylacetonate domains grown on H-terminated porous silicon at room temperature and their specific I-V behavior. AB - Porous silicon (PS) was incubated in an organic solution of metal acetylacetonates of Mn(acac)(3), Fe(acac)(3), Co(acac)(3), and Ni(acac)(2) (acac = MeCOCHCOMe) at room temperature. Crystal-like domains were found to be spontaneously self-assembled on PS surfaces by atomic force microscopy (AFM). Spectroscopic studies with attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS) revealed that the domains were grown from metal acetylacetonates. Current sensing atomic force microscopy (CSAFM) was used to measure the I-V curves of domains in nanoscale and specific step-jump currents on the manganese and cobalt acetylacetonate domains were surprisingly detected. PMID- 17134217 TI - A novel, sensitive potentiometric hydrocarbon sensor for high-vacuum applications. AB - A potentiometric device based on interfacing a solid electrolyte oxygen ion conductor with a thin platinum film acts as a robust, reproducible sensor for the detection of hydrocarbons in high- or ultrahigh-vacuum environments. Sensitivities in the order of approximately 5 x 10(-10) mbar are achievable under open circuit conditions, with good selectivity for discrimination between n butane on one hand and toluene, n-octane, n-hexane, and 1-butene on the other hand. The sensor's sensitivity may be tuned by operating under constant current (closed circuit) conditions; injection of anodic current is also a very effective means of restoring a clean sensing surface at any desired point. XPS data and potentiometric measurements confirm the proposed mode of sensing action: the steady-state coverage of Oa, which sets the potential of the Pt sensing electrode, is determined by the partial pressure and dissociative sticking probability of the impinging hydrocarbon. The principles established here provide the basis for a viable, inherently flexible, and promising means for the sensitive and selective detection of hydrocarbons under demanding conditions. PMID- 17134218 TI - Interactions of O(2) with Pd nanoparticles on alpha-Al(2)O(3)(0001) at low and high O(2) pressures. AB - The interaction of O(2) with small Pd particles (2-10 nm) supported on an alpha Al(2)O(3)(0001) single crystal under both ultrahigh vacuum (UHV) and high pressure conditions has been studied by temperature-programmed desorption (TPD), temperature-programmed low-energy ion scattering (TP-LEIS), and X-ray photoelectron spectroscopy (XPS). A low O(2) exposure (30 L) at 500 K leads to surface oxygen adatoms on the Pd nanoparticles, which desorb in TPD as O(2) in a peak at approximately 880 K. Surface O adatoms on the smallest Pd particles move to subsurface sites starting at 400 K, and they almost all move subsurface by approximately 750 K, desorbing mainly at considerably higher temperature. The dominant oxygen species above 700 K is subsurface, implying that it is more stable than oxygen adatoms on Pd. Exposures of the Pd nanoparticles to 25 Torr O(2) at 373-473 K readily convert the Pd to a species whose Pd XPS peak shifts by the same amount as the binding energy difference between bulk Pd and bulk PdO. We attribute this to PdO nanoparticles (or a thin film of PdO on or under the Pd for the larger particles). The decomposition of the PdO on these nanoparticles to Pd in an equilibrium O(2) pressure of 10-7 Torr does not occur until approximately 750 K, or approximately 200 K higher than the equilibrium decomposition of bulk PdO. This is attributed to the higher energy of Pd nanoparticles compared to bulk Pd and, for the larger particles, to the adhesion energy of the PdO film to the Pd, both of which stabilize the PdO on these Pd nanoparticles relative to bulk PdO. This PdO-like film on the larger particles may be similar to the ordered oxide thin film previously reported to form on Pd(111) but may also reside at the alpha-Al(2)O(3) interface and be partially stabilized by adhesion to this interface. PMID- 17134219 TI - Electrochemical fabrication of two-dimensional palladium nanostructures as substrates for surface enhanced Raman scattering. AB - Two-dimensional palladium (Pd) nanostructures have been fabricated by electrochemical deposition of Pd onto an indium tin oxide glass substrate modified with a thin flat film of polypyrrole or a nanofibril film of polyaniline. The experimental results demonstrated that the morphology of Pd nanoparticles strongly depended on the properties of conducting polymers and the conditions of electrochemical deposition. Two-dimensional nanostructures composed of flower-like (consisting of staggered nanosheets) or pinecone-like Pd nanoparticles were successfully synthesized. They can be used as substrates for surface-enhanced Raman scattering after partly decomposing the polymer components by heating in air, and the enhancement factor of the substrate composed of flower like Pd nanoparticles was measured to be as high as 105 for 4-mercaptopyridine. PMID- 17134220 TI - Methane oxidation mechanism on Pt(111): a cluster model DFT study. AB - The electronic energy barriers of surface reactions pertaining to the mechanism of the electrooxidation of methane on Pt (111) were estimated with density functional theory calculations on a 10-atom Pt cluster, using both the B3LYP and PW91 functionals. Optimizations of initial and transition states were performed for elementary steps that involve the conversion of CH(4) to adsorbed CO at the Pt/vacuum interface. As a first approximation we do not include electrolyte effects in our model. The reactions include the dissociative chemisorption of CH(4) on Pt, dehydrogenation reactions of adsorbed intermediates (*CH(x) --> *CH(x-1) + *H and *CH(x)O --> *CH(x-1)O + *H), and oxygenation reactions of adsorbed CH(x) species (*CH(x) + *OH --> *CH(x)OH). Many pathways were investigated and it was found that the main reaction pathway is CH(4) --> *CH(3) -> *CH(2) --> *CH --> *CHOH --> *CHO --> *CO. Frequency analysis and transition state theory were employed to show that the methane chemisorption elementary step is rate-limiting in the above mechanism. This conclusion is in agreement with published experimental electrochemical studies of methane oxidation on platinum catalysts that have shown the absence of an organic adlayer at electrode potentials that allow the oxidation of adsorbed CO. The mechanism of the electrooxidation of methane on Pt is discussed. PMID- 17134221 TI - Preparation of carbon-supported core-shell Au-Pt nanoparticles for methanol oxidation reaction: The promotional effect of the Au core. AB - Core-shell Au-Pt nanoparticles with intimate contact of Pt and Au were prepared by a displacement reaction without formation of monometallic Au nanoparticles. The Au-Pt nanoparticles were dispersed on carbon (Au@Pt/C) and were used to catalyze methanol electrooxidation in acidic solutions at room temperature. The core-shell nanostructure was confirmed by transmission electron microscopy and X ray photoelectron spectroscopy, and specific catalytic activities were evaluated by CO anodic stripping voltammetry in 0.5 M H(2)SO(4) and by cyclic voltammetry in 1 M CH(3)OH + 0.5 M H(2)SO(4). The Au@Pt/C catalyst demonstrated enhanced specific activity in methanol electrooxidation and showed multiple CO stripping peaks which were all negatively shifted with respect to a similarly prepared Ag@Pt/C catalyst. The activity enhancement is attributed to the presence of Au underneath a very thin Pt shell where electron exchange between Au and Pt had promoted the formation of active oxygen species on Pt, which facilitated the removal of inhibiting CO-like reaction intermediates. PMID- 17134222 TI - Trapping of charge carriers in colloidal particles of self-assembled films from TiO(2) and poly(vinyl sulfonic acid). AB - Self-assembled electrodes consisting of TiO(2) nanoparticles and poly(vinyl sulfonic acid) (PVS) were prepared by the layer-by-layer (LbL) technique. The electrostatic interaction between the TiO(2) nanoparticles and PVS allowed the growth of visually uniform multilayers of the composite, with high control of the thickness and nanoarchitecture. The electrochemical and chromogenic properties of these TiO(2)/PVS films were examined in an electrolytic solution of 0.5 M LiClO(4)/propylene carbonate. The presence of two intercalation sites was noted during the positive potential scan, and they were attributed to different mobilities of charge carriers. Several charge/discharge cycles demonstrated the trapping of charge carriers in the TiO(2) sites. The absorbance change associated with the oxidation of the trapping sites was attributed to electronic transitions involving energy states in the gap band formed due to the strong distortion of the TiO(2) host. Using the quadratic logistic equation (QLE), it was possible to analyze the electronic intervalence transfer from Ti(3+) to Ti(4+). Using the parameters obtained from this fitting, the amount of trapping sites in the LbL film was also determined. Electrochemical impedance spectroscopy (EIS) data gave the time constant associated with diffusion and the trapping sites. The diffusion coefficient of lithium ions changed from ca. 4.5 x 10(-13) cm(2) s(-1) to 3.0 x 10(-14) cm(2) s(-1) for all the potential range applied, indicating that PVS did not hinder the ionic transport within the LbL film. Finally, on the basis of the spectroelectrochemical data and scanning electron micrographs, the trapping effects were attributed to the colloidal particles of Li(0.55)TiO(2). PMID- 17134223 TI - Self-assembled monolayers of ferrocene-substituted biphenyl ethynyl thiols on gold. AB - Homogeneous and mixed [with biphenylthiol (BPT)] self-assembled monolayers (SAMs) of ferrocene-substituted biphenyl ethynyl thiols (Fc) were prepared on Au(111) substrates and characterized by several complementary spectroscopic techniques. The mixed films were fabricated either by subsequent immersion of the substrates into the BPT and Fc solutions or by immersion of the substrate into a mixed solution of BPT and Fc. The first procedure resulted in the preparation of high quality mixed SAMs, in which the Fc molecules were stochastically distributed in the BPT matrix and well-separated from each other. The portion of these molecules in such films could be precisely varied from ca. 7 to 42% by selection of the immersion time in the BPT solution. The films prepared from the mixed solution exhibited a phase separation between the Fc and BPT constituents. These films contained mostly the Fc molecules ( approximately 80-90%), showing, thus, a significant deviation from the relative content of the target molecules in the primary solution (a 1:1 ratio). This finding shows that the Fc molecules, when competing with BPT, preferably adsorb onto Au(111) substrate, suggesting a significant impact of the ferrocene groups onto the structure-building interactions responsible for molecular self-assembly. PMID- 17134224 TI - Photocatalytic activity of a Bi-based oxychloride Bi(3)O(4)Cl. AB - A Bi-based oxychloride Bi(3)O(4)Cl with a layered structure as a novel efficient photocatalyst was studied in the present paper. The compound synthesized by a solid-state reaction method has a band gap of 2.79 eV. The material possesses a fair visible-light-induced photocatalytic activity. Generally, the photocatalytic efficiency of Bi(3)O(4)Cl for degrading methyl orange (MO) is higher than that of anatase TiO(2) under UV light illumination. The dispersion of Ag over Bi(3)O(4)Cl leads to an obvious increase in the photocatalytic performance. The MO decolorization over Bi(3)O(4)Cl is mainly initiated by a photocatalytic process. The photocatalytic activity is discussed in close connection with the crystal structure and the electronic structure in details. PMID- 17134225 TI - Electrodeposition of poly(N-vinylcarbazole) at the three-phase junction. Formation of very different polymer structures. AB - Poly(N-vinylcarbazole) films can be deposited at the three-phase boundary when the organic phase contains only monomer, N-vinylcarbazole, while the aqueous phase contains supporting electrolyte. A cylindrical platinum microelectrode is immersed into the two-liquid system in such a way that a part of it is located in one liquid and the other part resides in the second liquid. The thickness of the reaction layer, or the width of the microelectrode zone where the polymer grows, depends on the kind of ions present in the aqueous phase and the time of the experiment. The structure of the deposited polymer may be very different and depends on the distance from the three-phase boundary and the type of the anion present in the aqueous phase. The key parameters here are the local electrode potential and the local concentration of the anions. The list of obtained polymer structures includes, among others, arrays of perfectly arranged deep oval channels and groups of microcrystals. The porosity of the polymer deposits increases with the distance from the aqueous phase. PMID- 17134226 TI - Transition between icosahedral and cuboctahedral nanoclusters of lead. AB - We have used ab initio methods to study the possible transition between icosahedral (ico) and cuboctahedral (fcc) structures in lead nanoclusters of sizes up to 309 atoms. Spontaneous fcc-to-ico transition in Pb(13) was observed in the ab initio molecular dynamics (MD) simulations at various temperatures. The transition path can be described predominantly by an angular variable s, which can, generally be applied to the similar transitions in clusters of larger sizes and was observed to follow the Mackay model. We have calculated the two dimensional energy surface that describes the transition in Pb(13) and found a barrierless fcc-to-ico transition path, which is consistent with the observed spontaneous transition in the ab initio MD simulations. The atomic displacements in the transition were identified as one of the vibrational eigenmodes of these two Pb(13) clusters. For clusters of larger sizes (Pb(n), where n = 55, 147, and 309), the possible transitions following similar paths were determined not to be barrierless and the sizes of the barriers were determined by the ab initio elastic band method. PMID- 17134227 TI - A theoretical investigation of the interactions between water molecules and ionic liquids. AB - Quantum chemical calculations have been used to investigate the interaction between water molecules and ionic liquids based on the imidazolium cation with the anions [Cl(-)], [Br(-)], [BF(4)(-)], and [PF(6)(-)]. The predicted geometries and interaction energies implied that the water molecules interact with the Cl( ), Br(-), and BF(4)(-0 anions to form X(-)...W (X = Cl or Br, W = H(2)O), 2X ...2W, BF(4)(-)...W, and W...BF(4)(-)...W complexes. The hydrophobic PF(6)(-) anion could not form a stable complex with the water molecules at the density functional theory (DFT) level. Further studies indicate that the cation could also form a strong interaction with the water molecules. The 1-ethyl-3 methylimidazolium cation (Emim(+)) has been used as a model cation to investigate the interaction between a water molecule and a cation. In addition, the interaction between the ion pairs and the water was studied by using 1-ethyl-3 methylimidazolium chloride (Emim x Cl) as a model ionic liquid. The strengths of the interactions in these categories follow the trend anion-W > cation-W > ion pair-W. PMID- 17134228 TI - Molecular dynamics simulations of polyelectrolyte-polyampholyte complexes. Effect of solvent quality and salt concentration. AB - Complexation between polyelectrolyte and polyampholyte chains in poor solvent conditions for the polyelectrolyte backbone has been studied by molecular dynamics simulations. In a poor solvent a polyelectrolyte forms a necklace-like structure consisting of polymeric globules (beads) connected by strings of monomers. The simulation results can be explained by assuming the existence of two different mechanisms leading to the necklace formation. In the case of weak electrostatic interactions, the necklace formation is driven by optimization of short-range monomer-monomer attraction and electrostatic repulsion between charged monomers on the polymer backbone. In the case of strong electrostatic interactions, the necklace structure appears as a result of counterion condensation. While the short-range attractions between monomers are still important, the correlation-induced attraction between condensed counterions and charged monomers and electrostatic repulsion between uncompensated charges provide significant contribution to optimization of the necklace structure. Upon forming a complex with both random and diblock polyampholytes, a polyelectrolyte chain changes its necklace conformation by forming one huge bead. The collapse of the polyelectrolyte chain occurs due to the neutralization of the polyelectrolyte charge by polyampholytes. In the case of the random polyampholyte, the more positively charged sections of the chain mix with negatively charged polyelectrolyte forming the globular bead while more negatively charged chain sections form loops surrounding the collapsed core of the aggregate. In the case of diblock polyampholyte, the positively charged block, a part of the negatively charged block, and a polyelectrolyte chain form a core of the aggregate with a substantial section of the negatively charged block sticking out from the collapsed core of the aggregate. In both cases the core of the aggregate has a layered structure that is characterized by the variations in the excess of concentration of monomers belonging to polyampholyte and polyelectrolyte chains throughout the core radius. These structures appear as a result of optimization of the net electrostatic energy of the complex and short-range attractive interactions between monomers of the polyelectrolyte chain. PMID- 17134229 TI - Structural and electronic characterization of the complexes obtained by the interaction between bare and hydrated first-row transition-metal ions (Mn(2+), Fe(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+)) and glycine. AB - The complexes formed by the simplest amino acid, glycine, with different bare and hydrated metal ions (Mn(2+), Fe(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+)) were studied in the gas phase and in solvent in order to give better insight into the field of the metal ion-biological ligand interactions. The effects of the size and charge of each cation on the organization of the surrounding water molecules were analyzed. Results in the gas phase showed that the zwitterion of glycine is the form present in the most stable complexes of all ions and that it usually gives rise to an eta(2)O,O coordination type. After the addition of solvation sphere, a resulting octahedral arrangement was found around Ni(2+), Co(2+), and Fe(2+), ions in their high-spin states, whereas the bipyramidal-trigonal (Mn(2+) and Zn(2+)) or square-pyramidal (Cu(2+)) geometries were observed for the other metal species, according to glycine behaves as bi- or monodentate ligand. Despite the fact that the zwitterionic structure is in the ground conformation in solution, its complexes in water are less stable than those obtained from the canonical form. Binding energy values decrease in the order Cu(2+) > Ni(2+) > Zn(2+) approximately Co(2+) > Fe(2+) > Mn(2+) and Cu(2+) > Ni(2+) > Mn(2+) approximately Zn(2+) > Fe(2+) > Co(2+) for M(2+)-Gly and Gly-M(2+) (H(2)O)(n) complexes, respectively. The nature of the metal ion-ligand bonds was examined by using natural bond order and charge decomposition analyses. PMID- 17134230 TI - Role of triple bond in 1,2-diphenylacetylene crystal: A combined experimental and theoretical study. AB - We have performed a combined experimental and theoretical study of the molecular system of 1,2-diphenylacetylene. The occurrence of two different geometries of the molecule in the crystal structure, one being planar and the other tilted by approximately 6 degrees , has been investigated in relation to the nature of the acetylenic linker. The experimental charge density analysis shows that the acetylenic linker exhibits a noncylindrical density reminiscent of the strong conjugation present in the molecule. The pi-orbitals of the acetylenic linker derived from density functional theory (DFT) calculations are found to sustain a variety of conjugation lengths between the phenyl rings, thereby giving flexibility to the molecule to arrange itself in various packing conformations in the crystal. It is interesting that the energy involved for such distortions is only kBT, allowing several polymorphic forms of the crystal structure as reported in the literature. The distortions entertained by the molecule and the corresponding changes in the charge density distribution and energy are all relevant to molecular electronics. PMID- 17134231 TI - Novel bifunctional viologen-linked pyrene conjugates: Synthesis and study of their interactions with nucleosides and DNA. AB - With the objective of developing efficient DNA oxidizing agents, a new series of viologen-linked pyrene conjugates with the general formula PYLnV(2+), having a different number of methylene spacer units (Ln) was synthesized, and their interactions with nucleosides and DNA have been investigated through photophysical and biophysical techniques. The viologen-linked pyrene derivatives PYL1V(2+) (n =equals; 1), PYL7V(2+) (n = 7), and PYL12V(2+) (n = 12) exhibited characteristic fluorescence emission of the pyrene chromophore centered around 380 nm but with significantly reduced yields when compared to those of the model compound PYL1Et(3)(+). The fluorescence quenching observed in these systems is explained through an electron-transfer mechanism based on a calculated favorable change in free energy (DeltaG(ET) = -1.59 eV), and the redox species characterized through laser flash photolysis studies. Intramolecular electron transfer rate constants (k(ET)) were calculated from the observed fluorescence yields, and the singlet lifetimes of the model compound and are found to decrease with increasing spacer length. The DNA binding studies of these systems through photophysical, chiroptical, and viscometric techniques demonstrated that these systems effectively undergo DNA intercalation with association constants (KDNA) in the range of 1.1-2.6 x 10(4) M(-1) and exhibit 2:1 sequence selectivity for poly(dG) x poly(dC) over poly(dA) x poly(dT). Photoactivation of these systems initiates electron transfer from the singlet excited state of the pyrene chromophore to the viologen moiety followed by an electron transfer from DNA to the oxidized pyrene. This results in the formation of stable charge-separated species such as radical cations of both DNA and reduced viologen as characterized by laser flash photolysis studies and subsequently the oxidized DNA modifications. These novel systems are soluble in buffer media, stable under irradiation conditions, and oxidize DNA efficiently and selectively through a cosensitization mechanism and hence can be useful as photoactivated DNA cleaving agents. PMID- 17134232 TI - Anthramycin-DNA binding explored by molecular simulations. AB - The anticancer drug anthramycin inhibits replication and transcription processes by covalently binding to DNA. Here, we use molecular simulations to investigate the interaction between this ligand and the dodecanucleotide d[GCCAACGTTGGC](2). We start from the X-ray structure of the adduct anthramycin-d[CCAACGTTG*G](2), in which the drug binds covalently to guanine.1 We focus on the noncovalent complexes between the oligonucleotide and the anhydro and hydroxy forms of the drug. Molecular dynamics (MD) simulations show that only the hydroxy form lies in front of the reactive center for the whole simulation ( approximately 20 ns), while the anhydro form moves inside the minor groove to the nearest base pair after approximately 10 ns. This sliding process is associated to both energetic and structural relaxations of the complex. The accuracy of our computational setup is established by performing MD simulations of the covalent adduct and of a 14-mer complexed with anhydro-anthramycin. The MD simulations are complemented by hybrid Car-Parrinello quantum mechanics/molecular mechanics (QM/MM) simulations. These show that in the noncovalent complexes the electric field due to DNA polarizes the hydroxy and, even more, the anhydro form of the drug as to favor a nucleophilic attack by the alkylating guanine. This suggests that the binding process may be characterized by a multistep pathway, catalyzed by the electric field of DNA. PMID- 17134233 TI - On the unusual stability of valence anions of thymine based on very rare tautomers: A computational study. AB - We characterized anionic states of thymine using various electronic structure methods, with the most accurate results obtained at the CCSD(T)/aug-cc-pVDZ level of theory followed by extrapolations to complete basis set limits. We found that the most stable anion in the gas phase is related to an imino-oxo tautomer, in which the N1H proton is transferred to the C5 atom. This valence anion, aT(c5)(nl), is characterized by an electron vertical detachment energy (VDE) of 1251 meV and it is adiabatically stable with respect to the canonical neutral nT(can) by 2.4 kcal/mol. It is also more stable than the dipole-bound (aT(dbs)(can)), and valence anion aT(val)(can) of the canonical tautomer. The VDE values for aT(dbs)(can)and T(val)(can) are 55 and 457 meV, respectively. Another, anionic, low-lying imino-oxo tautomer with a VDE of 2458 meV has a proton transferred from N3H to C5 aT(c5)(n3). It is less stable than aT(val)(can) by 3.3 kcal/mol. The mechanism of formation of anionic tautomers with the carbons C5 or C6 protonated may involve intermolecular proton transfer or dissociative electron attachment to the canonical neutral tautomer followed by a barrier-free attachment of a hydrogen atom to C5. The six-member ring structure of the anionic tautomers with carbon atoms protonated is unstable upon an excess electron detachment. Within the PCM hydration model, the low-lying valence anions become adiabatically bound with respect to the canonical neutral; becomes the most stable, being followed by aT(c5)(nl), aT(c5)(n3), aT(can), and aT(c5)(nl). PMID- 17134234 TI - Enzyme dynamics and tunneling enhanced by compression in the hydrogen abstraction catalyzed by soybean lipoxygenase-1. AB - A fully microscopical simulation of the rate-limiting hydrogen abstraction catalyzed by soybean lipoxygenase-1 (SLO-1) has been carried out. This enzyme exhibits the largest, and weakly temperature dependent, experimental H/D kinetic isotope effect (KIE) reported for a biological system. The theoretical model used here includes the complete enzyme with a solvation shell of water molecules, the Fe(III)-OH- cofactor, and the linoleic acid substrate. We have used a hybrid QM(PM3/d-SRP)/MM method to describe the potential energy surface of the whole system, and the ensemble-averaged variational transition-state theory with multidimensional tunneling (EA-VTST/MT) to calculate the rate constant and the primary KIE. The computational results show that the compression of the wild-type active site enzyme results in the huge contribution of tunneling (99%) to the rate of the hydrogen abstraction. Importantly, the active site becomes more flexible in the Ile553Ala mutant reactant complex simulation (for which a markedly temperature dependent KIE has been experimentally determined), thus justifying the proposed key role of the gating promoting mode in the reaction catalyzed by SLO-1. Finally, the results indicate that the calculated KIE for the wild-type enzyme has an important dependence on the barrier width. PMID- 17134235 TI - Interpreting DNA vibrational circular dichroism spectra using a coupling model from two-dimensional infrared spectroscopy. AB - Two-dimensional infrared spectroscopy was recently used to measure the vibrational couplings between carbonyl bonds located on DNA nucleobases (Krummel, A. T.; Mukherjee, P.; Zanni, M. T. J. Phys. Chem. B 2003, 107, 9165 and Krummel, A. T.; Zanni, M. T. J. Phys. Chem. B 2006, 110, 13991). Here, we extend the coupling model derived from these 2D IR experiments to simulate the vibrational absorption and vibrational circular dichroism (VCD) spectra of three double stranded DNA oligomers: poly(dG)-poly(dC), poly(dG-dC), and dGGCC. Using this model, we determine that the VCD spectrum of A-form poly(dG)-poly(dC) is dominated by interactions between stacked bases, whereas the coupling between base pairs and stacked bases carries equal importance in the VCD spectrum of B form poly(dG-dC). We also simulate the absorption and VCD spectra of dGGCC, which is a combination of A- and B-form configurations. These simulations give insight into the structural interpretation of VCD and absorption spectroscopies that have long been used to monitor DNA secondary structure and kinetics. PMID- 17134236 TI - Two-component polypeptides modeled with effective pair potentials. AB - We present Monte Carlo simulations performed within a model based on a set of distance-dependent effective potentials which are used to describe the interactions between a pair of distinct amino acids. These effective potentials are extracted from experimental correlation functions through the Ornstein Zernike equations and adequate closure approximations. We focus our attention on the sequences of two specific residues, namely, alanine and glycine. The studied sequences are (a) (Ala)(12)-(Gly)(4)-(Ala)(12) and (b) three interacting chains of alternating alanines and glycines (with five residues per chain). The resulting structures are combinations of known secondary structures. More importantly, we verify that our simulated structures are in thermodynamic equilibrium by means of an estimation of the density of states. PMID- 17134237 TI - Ultrafast electron transfer in photosynthesis: reduced pheophytin and quinone interaction mediated by conical intersections. AB - The mechanism of electron transfer (ET) from reduced pheophytin (Pheo(-)) to the primary stable photosynthetic acceptor, a quinone (Q) molecule, is addressed by using high-level ab initio computations and realistic molecular models. The results reveal that the ET process involving the (Pheo(-) + Q) and (Pheo + Q(-)) oxidation states can be essentially seen as an ultrafast radiationless transition between the two hypersurfaces taking place via conical intersections (CIs). According to the present findings, an efficient ultrafast ET implies that the Pheo- and Q move toward each other in a given preferential parallel orientation, reaching the most effective arrangement for ET at intermolecular distances (R) around 5-3 Angstrom, where the lowest CIs are predicted. Favored donor/acceptor interactions are related to orientations with some overlap between the lowest occupied molecular orbitals (LUMO) of the two systems, and they lead to state crossings at an earlier stage of the movement (larger R). Furthermore, when the topology of the interacting moieties does not make possible the LUMOs overlap, the corresponding diabatic potential energy curves do not intersect. Thus, it is anticipated that large scale motions, which are difficult to monitor experimentally, are actually occurring in the photosynthetic reaction centers of bacteria, algae, and higher plants, to fulfill the observed ultrafast ET processes. PMID- 17134238 TI - Structural disorder of the CD3zeta transmembrane domain studied with 2D IR spectroscopy and molecular dynamics simulations. AB - In a recently reported study [Mukherjee, et al. Proc. Natl. Acad. Sci. U.S.A. 2006, 103, 3528] we used 2D IR spectroscopy and 1-(13)C=(18)O isotope labeling to measure the vibrational dynamics of 11 amide I modes in the CD3zeta transmembrane domain. We found that the homogeneous line widths and population relaxation times were all nearly identical, but that the amount of inhomogeneous broadening correlated with the position of the amide group inside the membrane. In this study, we use molecular dynamics simulations to investigate the structural and dynamical origins of these experimental observations. We use two models to convert the simulations to frequency trajectories from which the mean frequencies, standard deviations, frequency correlation functions, and 2D IR spectra are calculated. Model 1 correlates the hydrogen-bond length to the amide I frequency, whereas model 2 uses an ab initio-based electrostatic model. We find that the structural distributions of the peptidic groups and their environment are reflected in the vibrational dynamics of the amide I modes. Environmental forces from the water and lipid headgroups partially denature the helices, shifting the infrared frequencies and creating larger inhomogeneous distributions for residues near the ends. The least inhomogeneously broadened residues are those located in the middle of the membrane where environmental electrostatic forces are weakest and the helices are most ordered. Comparison of the simulations to experiment confirms that the amide I modes near the C-terminal are larger than at the N-terminal because of the asymmetric structure of the peptide bundle in the membrane. The comparison also reveals that residues at a kink in the alpha-helices have broader line widths than more helical parts of the peptide because the peptide backbone at the kink exhibits a larger amount of structural disorder. Taken together, the simulations and experiments reveal that infrared line shapes are sensitive probes of membrane protein structural and environmental heterogeneity. PMID- 17134239 TI - Density functional theory study of the beta-carotene radical cation and deprotonated radicals. AB - The beta-carotene radical cation and deprotonated neutral radicals were studied at the density functional theory (DFT) level using different density functionals and basis sets: B3LYP/3-21G, SVWN5/6-31G*, BPW91/DGDZVP2, and B3LYP/6-31G**. The geometries, total energies, spin distributions, and isotropic and anisotropic hyperfine coupling constants of these species were calculated. Deprotonation of the methyl group at the double bond of the cyclohexene ring of the carotenoid radical cation at 5 or 5' produces the most stable neutral radical because of retention of the pi-conjugated system while less stable deprotonation at 9 or 9' and 13 or 13' of the chain methyl groups causes significant distortion of the conjugation. The predicted methyl hyperfine coupling constants of 13-16 MHz of the neutral radicals are in good agreement with the previous electron nuclear double resonance (ENDOR) spectrum of photolyzed beta-carotene on a solid support. DFT calculations on the beta-carotene radical cation in a polar water environment showed that the polar environment does not cause significant changes in the proton hyperfine constants from those in the isolated gas-phase molecule. DFT calculated methyl proton hyperfine coupling constants of less than 7.2 MHz are in agreement with those reported for the radical cation in photosystem II (PS II) and those found in the absence of UV light for the radical cation on a silica alumina matrix. PMID- 17134240 TI - Effect of salt identity on the phase diagram for a globular protein in aqueous electrolyte solution. AB - Monte Carlo simulations are used to establish the potential of mean force between two globular proteins in an aqueous electrolyte solution. This potential includes nonelectrostatic contributions arising from dispersion forces first, between the globular proteins, and second, between ions in solution and between each ion and the globular protein. These latter contributions are missing from standard models. The potential of mean force, obtained from simulation, is fitted to an analytic equation. Using our analytic potential of mean force and Barker Henderson perturbation theory, we obtain phase diagrams for lysozyme solutions that include stable and metastable fluid-fluid and solid-fluid phases when the electrolyte is 0.2 M NaSCN or NaI or NaCl. The nature of the electrolyte has a significant effect on the phase diagram. PMID- 17134241 TI - Calorimetric and dynamic light-scattering investigation of cationic surfactant- DNA complexes. AB - By means of combined calorimetric and dynamic light-scattering measurements, we have investigated the conformational behavior of DNA chains after thermal melting in the presence of a cationic surfactant at different concentrations, up to a surfactant-to-phosphate group molar ratio close to unity. Both the specific heat capacity, C(ex)(p) and the hydrodynamic radius R of the DNA chains provide support for the existence of two structural arrangements with different thermal stabilities, coexisting in the bulk solution. Although a component remains an elongated unfolded DNA chain originated in the thermal denaturation, the second component, consisting of DNA-surfactant complexes, assumes a compact structure with an average size of about 80 nm, whose thermal denaturation occurs at temperatures higher than 100 degrees C. PMID- 17134242 TI - SCRF-DFT and NMR comparison of tetracycline and 5a,6-anhydrotetracycline in solution. AB - A combination of structures, energies, and spectral data calculated using density functional theory (DFT) with experimental NMR data has been used to assign conformational equilibria for tetracycline and 5a,6-anhydrotetracycline in water at pH 1, 7, and 10 and in chloroform (5a,6-anhydrotetracycline) and methanol (tetracycline). The results suggest that tetracycline always prefers the extended conformation but that 5a,6-anhydrotetracycline exists in water as a mixture of the two conformers and in chloroform exclusively in the twisted conformation. The conformational equilibria are also shown to be pH dependent. PMID- 17134243 TI - Calculation of equilibrium compositions of systems of enzyme-catalyzed reactions. AB - When a system of enzyme-catalyzed reactions does not involve H(2)O as a reactant, the equilibrium composition at specified temperature, pH, and ionic strength can be calculated using the Mathematica programs equcalcc, which uses the conservation matrix, or equcalcrx, which uses the stoichiometric number matrix. When H(2)O is involved as a reactant, equcalcrx must be used because H(2)O is not in the stoichiometric number matrix. It is shown here that the use of equcalcrx is equivalent to using the further transformed Gibbs energy G" that eliminates oxygen from the conservation matrix. Calculations presented here show that the calculation of further transformed Gibbs energies of formation of reactants other than coenzymes can be avoided by using equcalcrx to calculate equilibrium concentrations. PMID- 17134244 TI - A highly photoconductive poly(vinylcarbazole)/2,4,7-trinitro-9-fluorenone Sol-Gel material that follows a classical charge-generation model. AB - Highly photoconductive properties are reported for organic-inorganic hybrid sol gel thin film materials composed of a classical poly(vinylcarbazole)/2,4,7 trinitro-9-fluorenone (PVK/TNF) polymeric mixture, entrapped in a SiO(2) matrix, whose pores have been chemically modified by organic functional groups. The highest photosensitivity obtained, 3.4 x 10(-10) cm Omega(-1) W(-1) at E 22 V microm(-)1, at the optimum molar ratio between the active components, TNF, PVK, and SiO(2), is in the range of the highest values ever reported for any PVK/TNF based classical photoconductive material. It is demonstrated that the PVK/TNF based sol-gel films follow Onsager's classical charge-generation model. The analysis of the photocurrent efficiency (Phi) of PVK/TNF-based sol-gel films by such a model provides the primary quantum yield of thermalized pair formation and the initial thermalized pair distance, phi(0) = 0.12 and r(0) = 66.1 Angstrom, respectively, for the optimized sample. As a result of Onsager's analysis, a notorious improvement of the photocurrent generation process was achieved for low TNF concentrations. PMID- 17134245 TI - Electron-transfer reactions with significant changes in structure. Unsymmetrical crowded ethylenes. AB - The electrochemical reduction mechanisms of xanthylideneanthrone, 6, thioxanthylideneanthrone, 7, 10-(diphenylmethylene)anthrone, 8, and 9 (diphenylmethylene)-9H-fluorene, 9, have been studied in dimethylformamide. The reduction of the first two compounds proceeds from folded forms of the neutral to twisted forms of the anion radical according to a square scheme. The data for reduction of 8 can be well accounted for by the same square scheme. However, one step reduction with concerted electron transfer and structural change cannot be ruled out. Compound 9, whose fluorene ring system cannot fold, exists only in twisted forms in the neutral, anion radical, and dianion. Consequently, there are no major changes in structure upon reduction, and the compound is reduced in two reversible steps with the second complicated by rapid loss of the dianion that is probably due to protonation by components of the medium. PMID- 17134247 TI - Converting gem-dimethyl groups into cyclopropanes via Pd-catalyzed sequential C-H activation and radical cyclization. AB - A novel route to the synthesis of cyclopropane derivatives is described. 1,1 Dimethyls in 2-(1,1-dimethylalkyl)dimethyloxazolines are first converted into 1,3 diiodide derivatives via Pd-catalyzed sequential C-H activation and then radically cyclized to provide 2-(1-alkylcylclopropyl)dimethyloxazolines. The use of EtOAc as a solvent is crucial for the diiodination of the functionalized substrates. [reaction: see text] PMID- 17134248 TI - A highly selective synthesis of dialkenyl sulfones via cross-metathesis of divinyl sulfone. AB - Catalytic cross-metathesis of commercial divinyl sulfone allowed direct access to novel (E)-alkenylvinyl sulfones and (E,E)-dialkenyl sulfones with excellent stereoselectivity. These compounds are useful building blocks, e.g., in the synthesis of substituted thiomorpholine 1,1-dioxide derivatives. [reaction: see text] PMID- 17134249 TI - An interesting issue of Diels-Alder selectivity discovered en route to 11-O debenzoyltashironin. AB - The hypervalent iodine-mediated oxidative dearomatization/Diels-Alder cascade was examined in the context of the natural product 11-O-debenzoyltashironin. Interestingly, the regioselectivity of the Diels-Alder reaction can be completely switched by changing the dienophile. Trapping of allyl alcohols during the oxidative dearomatization gives rise to the five-membered acetal, while trapping of allenyl alcohols results in the six-membered acetal. [reaction: see text] PMID- 17134250 TI - Selective reduction of peptide isothiazolidin-3-ones. AB - Isothiazolidinones are a rare but potentially important chemical moiety in biochemistry. We report the identification of several thiol, phosphinate, and carbon nucleophiles that form covalent adducts by addition to the sulfenamide sulfur. This reduction is selective for isothiazolidinones over similar peptide disulfides. We synthesized a coumarin-based thioacid nucleophile which shows a marked fluorescence increase after addition to an isothiazolidinone sulfenamide bond. [structure: see text] PMID- 17134251 TI - Rigid linear and star-shaped pi-conjugated 2,2':6',2"-terpyridine ligands with blue emission. AB - A series of rigid linear and star-shaped pi-conjugated 2,2':6',2"-terpyridine derivatives with the C-C single-, double-, and triple-bond linkages have been developed through the Suzuki, the Sonogashira, and the Wittig reactions. Such large rigid ligands exhibit strong blue emission in dilute solutions. [structure: see text] PMID- 17134252 TI - New procedure to mask the 2,3-pi bond of the indole nucleus and its application to the preparation of potent opioid receptor agonists with a Corynanthe skeleton. AB - Treatment of indole alkaloids with hypervalent iodine in the presence of ethylene glycol provides 2,3-ethylene glycol bridged adducts that could be converted into the original indoles under mild reductive conditions. This procedure, which involves masking of the reactivity of the indole nucleus at the beta-position, was utilized for the modification of the benzene ring of the indoline derivative and was applied to the preparation of potent opioid receptor agonists with the Corynanthe skeleton. [reaction: see text] PMID- 17134253 TI - Chaetominine, a cytotoxic alkaloid produced by endophytic Chaetomium sp. IFB E015. AB - Chaetominine (1), an alkaloidal metabolite with a new framework, was characterized from the solid-substrate culture of Chaetomium sp. IFB-E015, an endophytic fungus on the apparently healthy Adenophora axilliflora leaves. Its structure was determined by a combination of its spectral data and single-crystal X-ray diffraction analysis, with its absolute configuration elucidated by Marfey's method. Chaetominine was more cytotoxic than 5-fluorouracil against the human leukemia K562 and colon cancer SW1116 cell lines. [structure: see text] PMID- 17134254 TI - Synthesis of a phosphorus-containing hybrid porphyrin. AB - A phosphorus-containing hybrid porphyrin was successfully prepared via the BF3 promoted dehydrative condensation between sigma4-phosphatripyrrane and 2,5 bis[hydroxy(phenyl)methyl]thiophene. The NMR and UV-vis absorption spectra, electrochemical measurements, and DFT calculations revealed that the sigma3 P,N2,S-hybrid porphyrin exhibits high aromaticity as an 18pi-electron system in terms of both geometric and magnetic criteria. [structure: see text] PMID- 17134255 TI - Curtius rearrangement of aromatic carboxylic acids to access protected anilines and aromatic ureas. AB - The reaction of a chloroformate or di-tert-butyl dicarbonate and sodium azide with an aromatic carboxylic acid produces the corresponding acyl azide, presumably through the formation of an azidoformate. The acyl azide undergoes a Curtius rearrangement to form an isocyanate derivative which is trapped either by an alkoxide or by an amine to form the aromatic carbamate or urea. The reaction conditions are compatible with a variety of functional groups and allow the synthesis of a number of aniline derivatives containing alkyl, halide, nitro, ketone, ether, and thioether substituents. [reaction: see text] PMID- 17134256 TI - Acridinium salt based fluorescent and colorimetric chemosensor for the detection of cyanide in water. AB - A new, selective chemosensor has been developed to detect cyanide in water at micromolar concentrations. The acridinium salt used in this sensor system is prepared in a single step from an acridine orange base. Detection is based on the irreversible, 1:1 stoichiometric, nucleophilic addition of cyanide to the 9 position of the acridinium ion. This process induces a large decrease in fluorescence intensity and a marked color change. The selectivity of the system in aqueous media for CN- over other anions is remarkably high. Also, the sensitivity of both the fluorescence- and colorimetric-based assay is below the 1.9 microM suggested by the World Health Organization (WHO) as the maximum allowable cyanide concentration in drinking water. Thus, the chemodosimeter should be applicable as a practical system for the monitoring of CN- concentrations in aqueous samples. [structure: see text] PMID- 17134257 TI - Palladium-catalyzed regiocontrolled alpha-arylation of trimethylsilyl enol ethers with aryl halides. AB - Inter- and intramolecular arylations of trimethylsilyl enol ethers with aryl halides are accomplished regiospecifically in the presence of a palladium catalyst and tributyltin fluoride in refluxing benzene or toluene. The optimal catalyst system called for the use of Pd2(dba)3 and tri-tert-butylphosphine in ca. 1:2 ratio. Aryl iodides, bromides, and chlorides are all effective arylation partners in this reaction. [reaction: see text] PMID- 17134258 TI - Asymmetric radical addition of ethers to enantiopure N-p-toluenesulfinyl aldimines, mediated by dimethylzinc-air. AB - Asymmetric radical addition of ethers to enantiopure aromatic N-p-toluenesulfinyl aldimines has been achieved. The requisite radicals were generated by dimethylzinc-air. Lewis acid activation of the N-p-toluenesulfinyl aldimines followed by radical addition gives a mixture of sulfinamide and sulfonamide products. Subsequent treatment of the mixture with dry m-CPBA affords the sulfonamide product in enantiomerically enriched form. [reaction: see text] PMID- 17134259 TI - Shortcut synthesis of beta-cyclomannin from beta-cyclodextrin. AB - Beta-cyclomannin, a cyclic oligosaccharide consisting of seven alpha-D-mannosides connected together by the (1-->4) glycoside linkage, has been efficiently synthesized by the OsO4 oxidation of heptakis(2,3-didehydroxy)-beta-cyclodextrin which was prepared from beta-cyclodextrin by a five-step transformation. The novel cyclooligosaccharide not only showed water solubility high enough to meet the requirement for drug formulation but also demonstrated strong binding ability toward guest molecules. [reaction: see text] PMID- 17134260 TI - Stereocontrolled route to vicinal diamines by [3.3] sigmatropic rearrangement of allyl cyanate: asymmetric synthesis of anti-(2R,3R)- and syn-(2R,3S)-2,3 diaminobutanoic acids. AB - A stereocontrolled route via allyl 1,2-diols to vicinal diamines based on the [3.3] sigmatropic rearrangement of allyl cyanate has been developed. Our approach consists of two consecutive steps: stereoselective construction of allyl anti- and syn-1,2-diols followed by [1,3]-chirality transfer by sigmatropic rearrangement, which allow an access to anti-(2R,3R)- and syn-(2R,3S)-2,3 diaminobutanoic acids. [reaction: see text] PMID- 17134261 TI - Uncatalyzed and solvent-free multicomponent process for the synthesis of biphenyl 2-carbonitrile derivatives. AB - An innovative route to prepare a number of variously substituted new biphenyl derivatives is presented here. The protocol avoids the use of a catalyst, an organic solvent, and dry conditions. [reaction: see text] PMID- 17134262 TI - Enantioselective conjugate addition of a lithium ester enolate catalyzed by chiral lithium amides. AB - Mixed aggregates of chiral lithium amide and lithium ester enolate have been employed in the enantioselective conjugate addition on alpha,beta-unsaturated esters. Michael adducts were obtained in ee's up to 76% combining a lithium enolate and a chiral 3-aminopyrrolidine lithium amide. The sense of the induction was found to be determined by both the relative configuration of the stereogenic centers borne by the amide and the solvent in which the reaction was conducted. [reaction: see text] PMID- 17134263 TI - Vibralactone: a lipase inhibitor with an unusual fused beta-lactone produced by cultures of the basidiomycete Boreostereum vibrans. AB - The structure and absolute configuration of vibralactone (1) from the cultures of the Basidiomycete Boreostereum vibrans were established by spectroscopic methods and computational methods. Vibralactone, an unusual fused beta-lactone-type metabolite, was found to inhibit pancreatic lipase with an IC50 of 0.4 microg/mL. [structure: see text] PMID- 17134264 TI - A density functional that accounts for medium-range correlation energies in organic chemistry. AB - It has recently been pointed out that current density functionals are inaccurate for computing stereoelectronic effects and energy differences of isomerization reactions and isodesmic reactions involving alkanes; this has been interpreted as an incorrect prediction of medium-range correlation energies. This letter shows that the recently published M05-2X functional has good accuracy for all three of the recently highlighted problems and should be useful for a wide variety of problems in organic chemistry. [structure: see text] PMID- 17134265 TI - Copper(I)-catalyzed chlorine atom transfer radical cyclization reactions of unsaturated alpha-chloro beta-keto esters. AB - Copper(I) chloride catalyzed chlorine atom transfer radical cyclization reactions of a series of olefinic alpha-chloro beta-keto esters were investigated. It was found that alpha-dichlorinated beta-keto esters were suitable substrates; the chlorine transfer mono or tandem radical cyclization reactions catalyzed by CuCl complex with bis(oxazoline) or bipyridine proceeded smoothly in dichloroethane at room temperature or 80 degrees C, providing cyclic and bicyclic compounds in moderate to high yield. [reaction: see text] PMID- 17134266 TI - New synthetic protocols for the preparation of unsymmetrical bisindoles. AB - Novel unsymmetrical bisindoles were synthesized by a solvent-free C-C bond formation reaction under mild conditions. Starting from aziridines or hydroxyl precursors, indoles have been used as C-nucleophiles to form new pharmacologically interesting bisindoles via an electrophilic aromatic substitution pathway in good to excellent yields. [reaction: see text] PMID- 17134267 TI - Efficient synthesis of a C-analogue of the immunogenic bacterial glycolipid BbGL2. AB - Synthesis of a C-analogue of bacterial glycolipid BbGL2 is reported using Grignard reaction of in situ generated beta-galactosyl iodide and subsequent olefin cross metathesis reaction of C-vinyl galactoside as key steps. [reaction: see text] PMID- 17134268 TI - Rearrangement of 3,3-disubstituted indolenines and synthesis of 2,3-substituted indoles. AB - Synthesis of 2,3-substituted indoles from phenylhydrazine and alpha-branched aldehydes via rearrangement of 3,3-disubstituted indolenine intermediates is reported. [reaction: see text] PMID- 17134270 TI - Highly regioselective synthesis of 2,3-disubstituted indenes via a novel palladium-catalyzed cyclization reaction of propargylic carbonates with carbon nucleophiles. AB - Palladium-catalyzed reaction of propargylic carbonates with carbon nucleophiles offers an efficient, direct route to highly substituted indenes. The reaction conditions and the scope of the process are examined, and a possible mechanism is proposed. [reaction: see text] PMID- 17134269 TI - Highly efficient and selective synthesis of conjugated triynes and higher oligoynes of biological and materials chemical interest via palladium-catalyzed alkynyl-alkenyl coupling. AB - Iteration of a Pd-catalyzed reaction of alkynyl- and oligoynylzincs with (E) ICH=CHCl followed by metalation-termination with electrophiles(E) has provided a linear route to conjugated tri- and tetraynes, and Pd-catalyzed monoalkynylation of 1,1-dibromoenynes accompanied by dehydrobromination has provided a convergent route to conjugated tri- , tetra- , and pentaynes. Both display unprecedented high efficiency and selectivity. [reaction: see text] PMID- 17134271 TI - Efficient one-pot, two-step, microwave-assisted procedure for the synthesis of polysubstituted 2-aminoimidazoles. AB - A microwave-assisted, one-pot, two-step protocol was developed for the construction of polysubstituted 2-aminoimidazoles. This process involves the sequential formation of imidazo[1,2-a]pyrimidinium salts from readily available 2 aminopyrimidines and alpha-bromocarbonyl compounds, followed by opening of the pyrimidine ring with hydrazine. [reaction: see text] PMID- 17134272 TI - Synthesis of glyco(lipo)peptides by liposome-mediated native chemical ligation. AB - Although native chemical ligation (NCL) is emerging as a powerful method for the assembly of (glyco)peptide building blocks, its applicability is reduced when peptide segments are poorly soluble in aqueous buffer. We have found that incorporating reactants in liposomes allows NCL of lipophilic peptides and lipopeptides. Furthermore, the reaction rates of liposome-mediated NCL are higher than traditional reaction conditions resulting in improved yields. [reaction: see text] PMID- 17134273 TI - Isocyanide addition to pyridinium salts. Efficient entry into substituted nicotinonitrile derivatives. AB - The addition of isocyanides to pyridinium salts is studied. The process takes place efficiently when a carboxamido group is present in the 3 position of the pyridine ring. The outcome of the reaction involves the stabilization of the nitrilium intermediate by the amide, which suffers a mild dehydration, leading regioselectively to beta-cyano-gamma-carbamoyl-1,4-dihydropyridines. In this way, a variety of nicotinamide derivatives were carbamoylated. Extension to quinolinium, isoquinolinium, and N-acylpyridinium salts is also reported. [reaction: see text] PMID- 17134274 TI - TiCl4 induced anti-Markovnikov rearrangement. AB - Stereoisomeric bicyclic tert-alcohols afforded identical ring-expansion products via cationic anti-Markovnikov rearrangement from perpendicular tert-cations into identical six-membered ring secondary cations by the treatment with TiCl4. These results provide evidence that the reaction takes place by the cationic stepwise mechanism. [reaction: see text] PMID- 17134275 TI - Bidirectional and colorimetric recognition of sodium and potassium ions. AB - Host 1 based on the phenolphthalein skeleton and two crown ether moieties demonstrated opposite behaviors toward sodium and potassium cations caused by bidirectional complexation. [reaction: see text] PMID- 17134276 TI - Convergent assembly of polycyclic ethers via acyl radical addition to unactivated enol ether. AB - A new convergent strategy for assembling 6/6- and 6/7-fused ether ring systems was developed. The key features in our method include Ag+-promoted facile formation of chemically labile enol ether from O,S-acetal and addition of an acyl radical to unactivated enol ether to cyclize a six- or seven-membered ether ring. [reaction: see text] PMID- 17134277 TI - Two convergent routes to the left-wing fragment of ciguatoxin CTX3C using O,S acetals as key intermediates. AB - Ciguatoxins, principal causative toxins of ciguatera seafood poisoning, are large ladderlike polycyclic ethers. Here, we report two convergent routes to synthesis of the multiolefinic left half of ciguatoxins based on a newly developed acyl radical strategy. Remarkably, only 13 steps from the monocyclic E-ring were required to construct the left wing. [reaction: see text] PMID- 17134278 TI - Nonenzymatic oligomerization of RNA by TNA templates. AB - Cytosine TNA promotes nonenzymatic, template-directed oligomerization of complementary activated rGMP, leading to selective and efficient formation of RNA products. This process models "genetic takeover" of a pre-RNA by RNA. [reaction: see text] PMID- 17134279 TI - Synthesis and properties of a fluorene-capped isotruxene: a new unsymmetrical star-shaped pi-system. AB - An unsymmetrical star-shaped pi-system (1) with an isotruxene core and three fluorene arms has been synthesized, and its photophysical, electrochemical, and thermochemical properties have been investigated and compared with the corresponding symmetrical truxene derivatives 2a-d (Kanibolotsky, A. L.; Berridge, R.; Skabara, P. J.; Perepichka, I. F.; Bradley, D. D. C.; Koeberg, M. J. Am. Chem. Soc. 2004, 126, 13695-13702). Stronger electronic couplings between the arms in 1 vs 2a-d lead to lower optical band gap, larger splitting in oxidation potentials, and superior thermostability. [structure: see text] PMID- 17134280 TI - Carbocations in action. Design, synthesis, and evaluation of a highly acid sensitive naphthalene-based backbone amide linker for solid-phase synthesis. AB - The design, synthesis, and properties of an extremely acid-labile backbone amide linker based on a regiospecifically substituted tetraalkoxy naphthaldehyde core are presented. This handle enables cleavage of peptide backbone amides (secondary amides) off a solid support using as little as 0.5% TFA in CH2Cl2. This proceeds without cleavage of tert-butyl ethers and tert-butyl esters. The design is based on a DFT study that predicted the most stabile alkoxy-substituted methyl naphthyl carbocation. [structure: see text] PMID- 17134281 TI - Enantioselective synthesis and selective monofunctionalization of (4R,6R)-4,6- dihydroxy-2,8-dioxabicyclo[3.3.0]octane. AB - An efficient, enantioselective synthesis of a disubstituted bis-THF scaffold 5 is described, as well as an efficient differentiation of the 1,3-diol unit. [reaction: see text] PMID- 17134282 TI - Optimized synthesis of hydrogen-bond surrogate helices: surprising effects of microwave heating on the activity of Grubbs catalysts. AB - This manuscript discusses microwave-assisted solid-phase synthesis of hydrogen bond surrogate based alpha-helices and analogues by ring-closing metathesis (RCM). Microwave-mediated RCM allows access to a greater variety of amino acid residues in the macrocycles in shorter reaction times and higher yields compared to conventional heating. Surprisingly, we discovered that the Grubbs II catalyst is highly active under the influence of microwaves but catalytically dead under oil-bath conditions for the metathesis of these peptide bisolefins. [reaction: see text] PMID- 17134283 TI - Palladium-catalyzed oxidative activation of arylcyclopropanes. AB - Palladium chloride-catalyzed intramolecular activation of electroneutral cyclopropane derivatives results in cleavage of the cyclopropane ring followed by formation of heterocyclic derivatives. Phenols, carboxylic acids, and amide groups were considered as substituents ortho to the cyclopropane ring in this catalytic activation chemistry. The regioselectivity observed in the case of amide-containing substrates was different from that of carboxylic acid-containing substrates, ruling out simple cyclopropane isomerization followed by a Wacker oxidation as the mechanistic pathway. [reaction: see text] PMID- 17134284 TI - Concise strategy to the core structure of the macrolide queenslandon. AB - The fully functionalized core structure of the macrolactone queenslandon was prepared using a novel strategy consisting of a glycolate aldol reaction and hydroboration of the derived enol ether 17 followed by Suzuki cross-coupling with an iodostyrene. After conversion of the cross-coupling product to the seco acid 22, Mitsunobu macrolactonization and protecting group manipulations led to the queenslandon model 5. [structure: see text] PMID- 17134285 TI - Regioselective synthesis of substituted pyrroles: efficient palladium-catalyzed cyclization of internal alkynes and 2-amino-3-iodoacrylate derivatives. AB - The first efficient and regioselective palladium-catalyzed cyclization of internal alkynes and 2-amino-3-iodoacrylates to give moderate to excellent yields of highly functionalized pyrroles has been developed. This approach is applicable to a range of alkynes and affords the deacylated pyrrole under reaction conditions for most substrates. [reaction: see text] PMID- 17134286 TI - 180 degree unidirectional bond rotation in a biaryl lactone artificial molecular motor prototype. AB - A bifunctional biaryl lactone has been synthesized that should be capable of iterative unidirectional aryl-aryl bond rotation via: (1) a diastereoselective lactone ring opening, (S)-1 to (P,S)-2 or (M,S)-2; (2) a chemoselective lactonization, (P,S)-2 or (M,S)-2 to (S)-3; and (3) a chemoselective hydrolysis, (S)-3 to (S)-1. Preliminary results of a racemic sample have indicated unidirectional 180 degrees rotation with very high directional selectivity per individual artificial molecular motor molecule through the first two steps of this sequence. [reaction: see text] PMID- 17134287 TI - Three distinct reactions of 3,4-dihydroisoquinolines with azlactones: novel synthesis of imidazoloisoquinolin-3-ones, benzo[a]quinolizin-4-ones, and benzo[d]azocin-4-ones. AB - A facile and direct synthetic entry to tricyclic imidazoloisoquinolin-3-ones and benzo[a]quinolizin-4-ones is reported based on the ring annulation of 1 unsubstituted and 1-substituted dihydroisoquinolines with azlactones under neutral conditions in a one-step procedure. Bicyclic 2,3-dihydrobenzo[d]azocin-4 ones were also prepared using simple azlactone and 1-substituted dihydroisoquinolines in a one-pot reaction. [reaction: see text] PMID- 17134288 TI - (2R)- and (2S)-3-fluoroalanine and their N-methyl derivatives: synthesis and incorporation in peptide scaffolds. AB - A convergent synthetic methodology has been developed to access both (2S)- and (2R)-3-fluoroalanine and their corresponding N-methyl analogues, in optically pure form, through a common oxazolidinone intermediate that can be obtained from L- or D-serine. In addition, a procedure for incorporation of these unnatural amino acids in peptide scaffolds is also disclosed herein that minimizes the occurrence of beta-elimination during amide bond formation. [reaction: see text] PMID- 17134289 TI - Highly active in situ catalysts for anti-Markovnikov hydration of terminal alkynes. AB - The anti-Markovnikov hydration of terminal alkynes to give aldehydes is catalyzed by complexes derived in situ from air-stable [CpRu(eta6-naphthalene)]PF6 (C) and 6-aryl-2-diphenylphosphinopyridines (L). Ligands L are readily available from a modular synthesis. Increasing the size of the ligand C-6 aryl group in the order R = Ph < mesityl < 2,4,6-triisopropylphenyl < (2,4,6-triphenyl)phenyl gave hydration catalysts of highest known activity. [reaction: see text] PMID- 17134290 TI - Total synthesis of (+/-)-hedychenone: trimethyldecalin terpene systems via stepwise allenoate diene cycloaddition. AB - The total synthesis of hedychenone 1 is described. The cycloaddition of the hindered diene 2 and the allenecarboxylate 3 has been shown conclusively to proceed via the [2+2] cycloadduct 5 to give a 2:1 mixture of the desired formal Diels-Alder adducts, the exo and endo isomers 4xn and is thus a stepwise [4+2] cycloaddition. The exo isomer 4x was converted in four steps (reduction, oxidation, olefination, and desilylation) into hedychenone 1. [reaction: see text] PMID- 17134291 TI - Radical addition to 1,4-benzoquinones: addition at O- versus C-atom. AB - Addition of alkyl radicals generated from B-alkylcatecholboranes onto 1,4 benzoquinones leads to substituted hydroquinones in good overall yields. Formation of aryl ethers via a unique radical addition to the oxygen atom of the enone system is the main reaction when bulky secondary and tertiary alkyl radicals are used. Less hindered secondary and primary radicals give the expected 1,4-conjugate addition products. [reaction: see text] PMID- 17134292 TI - Thermolysis of isomigrastatin and its congeners via [3,3]-sigmatropic rearrangement: a new route to the synthesis of migrastatin and its analogues. AB - Thermolysis of isomigrastatin (1) under neat heating conditions afforded migrastatin (1a). The reaction is proposed to proceed via a concerted [3,3] sigmatropic rearrangement by which ring expansion is achieved regio- and enantiospecifically. The general applicability of this reaction was demonstrated with six additional isomigrastatin congeners (3-8), providing a new route to the synthesis of migrastatin analogues (3a-8a). [reaction: see text] PMID- 17134293 TI - Environment-sensitive fluorophore emitting in protic environments. AB - The unusual fluorescence properties of 8-methoxy-4-methyl-2H-benzo[g]chromen-2 one (1) are described. The fluorophore 1 is almost nonfluorescent in aprotic solvent (e.g., fluorescence quantum yield Phi(f) < 0.0003 in n-hexane), whereas it strongly fluoresces at long wavelengths (>450 nm) in protic solvent (e.g., Phi(f) = 0.21 in methanol). The fluorophore 1 also shows good applicability in developing a new fluorogenic (fluorescent "off-on") sensor. [structure: see text] PMID- 17134294 TI - First enantioselective total synthesis and structure determination of the anthrapyran metabolite gamma-indomycinone. AB - The first total synthesis of (R)-gamma-indomycinone has been achieved which allowed the determination of the configuration of the stereogenic center of natural gamma-indomycinone as (S). The approach stands out for its generality and efficiency. [reaction: see text] PMID- 17134295 TI - Nitrene equivalent mediated metal-free ring expansions of alkylidenecyclopropanes and an alkylidenecyclobutane. AB - The ring expansion of small-ring compounds provides a powerful method for the construction of various cyclic compounds. Herein, nitrene equivalent mediated metal-free ring expansions of alkylidenecyclopropanes (ACPs) and an alkylidenecyclobutane (ACB) were described. In this synthesis, a series of aryl substituted cyclobutylidene and cyclopentylidene hydrazine derivatives were obtained under mild conditions in moderate to good yields. [reaction: see text] PMID- 17134296 TI - Practical approach to alpha- or gamma-heterosubstituted enoic acids. AB - The reaction of alkenyl trichloromethyl carbinols with various nucleophiles under protic basic conditions reveals that mercaptans participate by alpha-substitution (S(N)2) of the intermediate alkenyl gem-dichloroepoxides. Conversely, hydroxide results in preferential gamma-substitution with stereoselective allylic transposition (S(N)2'). Regioselectivity with alkoxides depends upon the level of alkene substitution. [reaction: see text] PMID- 17134297 TI - Folded metallodendrons: shell-selective metal coordination and conformational properties. AB - The synthesis, structure, and conformational properties of folded metallodendrons are described. The enhanced coordination ability of oxazoline-functionalized pyridyl-2,6-dicarboxamides, compared with pyridyl-2.6-diarylamides, permits unique metals to be installed at the peripheral and focal shells of the dendron. Circular dichroism (CD) studies and X-ray crystallography indicate that the intrinsic helical preference of the parent dendron is not perturbed by coordination. However, the CD spectra become insensitive to temperature due to an increase in structural rigidity that occurs upon metal coordination. [structure: see text] PMID- 17134298 TI - One-pot synthesis of 2,3-dihydro-pyrrolopyridinones using in situ generated formimines. AB - A novel one-pot methodology is described for the synthesis of functionalized pyrrolopyridinones using in situ generated formimines and an ortho-lithiated pyridinecarboxamide species. Depending on the reaction conditions, this procedure allows versatile access to aminomethylated pyridinecarboxamides, 2,3-dihydro pyrrolopyridinones, or 1,1-dialkylated 2,3-dihydro-pyrrolopyridinone derivatives. [reaction: see text] PMID- 17134299 TI - Cationic dithieno[3,2-b:2',3'-d]phospholes: a new building block for luminescent, conjugated polyelectrolytes. AB - Cationic dithieno[3,2-b:2',3'-d]phospholes are accessible very efficiently by methylation of the phosphorus center. Further functionalization with bromo substituents in 2,6-positions affords a polymerizable monomer that can be copolymerized with a difunctionalized fluorene in a Suzuki-Miyaura-type cross coupling protocol. The monomers as well as the resulting conjugated polyelectrolyte based on the phospholium units show very intriguing photoluminescence properties, even in the solid state. [structure: see text] PMID- 17134300 TI - Enantioselective synthesis of functionalized medium-sized oxacycles. AB - Enantioselective routes to functionalized, seven-, eight-, and nine-membered oxacycles that are amenable to further elaboration have been developed. Salient features of the methodology include highly diastereoselective and regioselective transformations of an ephedrine-derived epoxy morpholinone to functionalized precursors of the oxacycles. The ephedrine scaffold exerts remote stereocontrol in the functionalization of the appended oxacycle. [reaction: see text] PMID- 17134301 TI - Exploring biosynthetic relationships among furanocembranoids: synthesis of (-) bipinnatin J, (+)-intricarene, (+)-rubifolide, and (+)-isoepilophodione B. AB - The asymmetric total synthesis of (-)-bipinnatin J and its conversion into (+) intricarene through a transannular 1,3-dipolar cycloaddition is described. In addition, the conversion of (-)-bipinnatin J into (+)-rubifolide and (+) isoepilophodione B is reported. Biosynthetic relationships among furanocembranoids and the possible role of 1,3-dipolar cycloadditions in biosynthesis are discussed. [reaction: see text] PMID- 17134302 TI - Gold-catalyzed synthesis of substituted tetrahydronaphthalenes. AB - We report a gold-catalyzed benzannulation of 3-hydroxy-1,5-enynes to generate tetrahydronaphthalenes. This mild process proves to be an effective method to synthesize various metasubstituted aromatic rings in good yields. [reaction: see text] PMID- 17134303 TI - Isocorannulenofuran: a versatile building block for the synthesis of large buckybowls. AB - Isocorannulenofuran, synthesized in two steps from accessible bromocorannulene, gives Diels-Alder adducts with benzynes which can be deoxygenated to produce large polycyclic aromatic hydrocarbons (PAHs) combining the bowl-shaped corannulene subunit with planar fragments. [reaction: see text] PMID- 17134304 TI - S-chiral sulfinamides as highly enantioselective organocatalysts. AB - Easily accessible chiral sulfinamide 2 has been developed as the first highly efficient and enantioselective organocatalyst relying solely on a chiral sulfur center for stereochemical induction. In the presence of 20 mol % of 2, a broad range of N-aryl ketimines 1 were reduced by trichlorosilane to produce amines 3 in high yield and enantioselectivity. [reaction: see text] PMID- 17134306 TI - [Management of congestive heart failure by internist and advantages of clinical investigation]. PMID- 17134307 TI - [Clinical and therapeutic characteristics in patients with heart failure cared for in an internal medicine department]. AB - INTRODUCTION: The Heart Failure (HF) is a very common clinic syndrome in the clinical practise, and a great sanitary social problem. In Spain, it is the main cause of hospitalization in Internal Medicine Services in patients over 65 years old, due to its high comorbidity, in many cases. OBJECTIVE: To determine the profile of admitted patients with HF to an Internal Medicine Service. MATERIAL AND METHODS: It is a retrospective and descriptive study. The following variables were analysed: sex, age, cause of admission, aetiology, risk factors, Framingham diagnostic criteria, type of HF, treatment-on-released, average stay, number of readmissions and the type of ventricular dysfunction (echocardiography). Chi square test of Pearson or Fisher test were used as statistical methods. P < 0.05 was considered to be statistically relevant. RESULTS: 104 patients were analysed. The percentage of HF was 13.76%. 52.88% were men with an average age of 72.16 +/- 17.78 year. Average age for women was 82.92 +/- 7.24 year. The main cause of admission was dysnea (74%). Hypertension was the principal risk factor (70.13%). When it comes to aetiology, HTA (68.57%) was the main one. Global and left HF (33.65% either) were the most common type of HF. 5.17% of women and 20.69% of men suffered from systolic dysfunction (p < 0.023). 32.76% of women and 41.38% of men had diastolic dysfunction. The base treatment-on-release for the majority of patients consisted of diuretics and digoxin. Patients stayed 15.49 days on average, and the number of readmitted ones came up to 26.67%. CONCLUSIONS: The diagnosis and treatment of HF in an Internal Medicine Service is acceptable. It will be very important to promote the use of beta blockers and angiotensina converting enzyme inhibitors. PMID- 17134308 TI - [Association patterns of risk factors or cardiovascular disease, in a high risk Spanish population of 2,264 patients]. AB - INTRODUCTION, BASES AND AIMS: The aim of this study is to present the different association patterns between cardiovascular disease risk factors and target organs lesion due to arterial hypertension (AH), and to establish the possible differences by gender and age, and compare our findings with the existing references in the medical literature. MATERIAL AND METHODS: Nationwide croo sectional study with 2264 consecutive patients in the Outpatient Clinic of Internal Medicine 18 years of age or older and with high cardiovascular risk taken from the CIFARC (Integral Control of High Risk Cardiovascular Factors) study run by the SEMI (Spanish Internal Medicine Association) cardiovascular Risk Group. RESULTS AND DISCUSSION: We have obtained statistically significant correlations between: Raised levels of total cholesterol and LDL cholesterol, low HDL cholesterol and smoking habit. LDL cholesterol with Family History of early cardiovascular events (hereinafter FH) and ischemic cardiopathy. Low HDL cholesterol with FH. Smoking habit with FH. Left ventricular hypertrophy (LVH) with raised levels of proteinuria and creatinine, atheromatosis, retinopathy I II, ischemic cardiopathy, and diabetic nephropathy. Ischemic cardiopathy with LVH, diabetic nephropathy, peripheral vascular disease (PVD) and age. Proteinuria with raised levels of creatinine, atheromatosis, retinopathy I-II, nephropathy and severe retinopathy. Retinopathy I-II with raised levels of creatinine, atheromatosis, nephropathy, PVD and severe retinopathy. Raised levels of creatinine with reduced levels of HDL cholesterol, proteinuria, atheromatosis, retinopathy I-II, nephropathy and PVD. FH inversely with age: patients with FH consult a doctor an average of three years earlier than patients who do not experience FH. Atheromatosis with LVH, raised levels of proteinuria and creatinine, retinopathy I-II, cerebrovascular disease, nephropathy, PVD and age. PVD with raised levels of creatinine, atheromatosis, retinopathy I-II, and ischemic cardiopathy. Cerebrovascular disease with atheromatosis, age and weight. Severe retinopathy with proteinuria, nephropathy, and retinopathy I-II. Diabetic nephropathy with raised levels of proteinuria and creatinine, atheromatosis, retinopathy I-II, ischemic cardiopathy, and severe retinopathy. Age with raised creatinine, atheromatosis, ischemic cardiopathy, and cerebrovascular disease. With respect to gender, we found differences in most of the studied variables. CONCLUSIONS: There exist different association patterns between the studied variables. PMID- 17134309 TI - [Use and abuse of intravenous catheters in conventional hospital wards]. AB - BACKGROUND: Information regarding the use intravenous catheters (IVC) in conventional hospital units and its consequence in terms of intravenous cateter related bacteremia (ICRB) is scarce. OBJECTIVES: To evaluate the use of IVC in patients admitted in conventional wards of a general hospital and to measure IVCRB incidence in such patients. METHODS: We evaluated during one week IVC use in adult patients admitted in 12 de Octubre Hospital and we calculated la incidence density of ICRB. RESULTS: We evaluated the clinical charts of 731 patients (284 from medical wards and 447 from surgical wards), of which 338 (46.2%) had a peripheral VC inserted and 63 (8.6%) a central IVC. Central IVC had been inserted for a mean time 11.5 days globally (CI 95% 5.57-17.42), being 28.3 in medical wards and 8.32 days in surgical wards (p = 0.2). In 27.7 % of the patients with IVC intravenous antimicrobials was the only reason for the use of such catheters in spite of adequate oral tolerance in 30 % of the patients with central IVC an specific note explaining the reason for implanting such catheter was lacking in the clinical chart. IVCRB was detected in 12/401 patients (3%). The incidence density of IVCRB in central IVC was 8.28 per 1000 catheter-days. CONCLUSIONS: There are some aspects that could be clearly improved regarding the prevention of IVCRB, mostly in the indications, the excess of time those catheters are kept implanted and in the lost chances for catheter withdrawal when switch-therapy could be performed. PMID- 17134310 TI - [Neurohormonal blockade in patients with heart failure at discharge from hospital]. AB - With the objective to value the degree of fulfillment of the pharmacological processing in patients with heart failure was designed the study of therapeutic adherence in heart failure (ATICA). During the period of inclusion educational, social, demographic data have been obtained, personal antecedents, analytic and physical exploration of the patients. These data mentioned are the ones that are reflected in the present pilot study. The total of patients included is 554, the majority of which are women, the middle ages is advanced and they present a fraction of eyeccion conserved in more than the half. In the therapeutic plan only was collected information of groups with neurohormonal action and of great recognition in the processing of the heart failure. The most utilized pharmacological group are the IECA, nevertheless medicines as the espirolactona or the betablocker continue showing a worrying degree of infrautilizacion, although they present a greater prescription that in other series. PMID- 17134311 TI - [Epstein-Barr virus infection and acute cholestatic hepatitis]. AB - Epstein-Barr virus (EBV) is a herpesvirus whose only reservoir host is the human. It is transmitted by oropharyngeal secretions. Primary EBV infection is usually asymptomatic, but sometimes it causes infectious mononucleosis with fever, lymphadenopathies, splenomegaly and pharyngitis. Acute infection is diagnosed by serology (heterophile or specific antibodies). Immunofluorescence and molecular biologic techniques may be used to demonstrate the presence of EBV in biopsy specimens. Mild and transient elevations of serum aminotransferases are common, thus liver biopsy is usually not necessary to confirm the diagnosis. Severe cholestasis is rare (5%). We describe a patient with cholestatic hepatitis and acute EBV infection with atypical lymphocytes and positive anti-VCA IgM. The patient had taken drugs (ibuprofen, paracetamol and valerian). The bad evolution of the patient, the history of exposure to drugs, and the few cases of cholestatic hepatitis due to EBV infection reported, led us to consider liver biopsy. Molecular biologic techniques confirmed the presence of EBV in liver tissue however histologic features did not exclude the toxic aetiology or the concomitant effect of drugs and EBV infection. PMID- 17134312 TI - [Non-bacterial thrombotic endocarditis and lung neoplasm]. AB - A woman with the final diagnosis of non-bacterial thrombotic endocarditis associated to pulmonary adenocarcinoma is reported. She had had repetitive embolic strokes as the first manifestation, with vegetations on the mitral valve and without signs of acute infectious disease. The case is interesting because is an usually case of nonbacterial thrombotic endocarditis and the importance of an early diagnostic. PMID- 17134313 TI - [Acrocyanosis as a form of presentation of progressive systemic sclerosis]. AB - Acrocyanosis is an infrequent entity characterized by persistent and symmetrical erythrocyanotic discoloration of the hands, feet and face, which is not preceded by episodes of previous paleness. Acrocyanosis can be secondary to a variety of underlying causes, but is uncommon as a form of presentation of systemic sclerosis. We present a new clinical case of this disease and discuss its etiopathogenic, clinical, diagnostic and therapeutic characteristics; in addition to, we seek to distinguish this term from others that might be confused in the literature. PMID- 17134314 TI - [Ethics conflicts in learning and teaching medical specialities]. PMID- 17134315 TI - [Myocarditis from clozapine]. PMID- 17134316 TI - Mean-driven and fluctuation-driven persistent activity in recurrent networks. AB - Spike trains from cortical neurons show a high degree of irregularity, with coefficients of variation (CV) of their interspike interval (ISI) distribution close to or higher than one. It has been suggested that this irregularity might be a reflection of a particular dynamical state of the local cortical circuit in which excitation and inhibition balance each other. In this "balanced" state, the mean current to the neurons is below threshold, and firing is driven by current fluctuations, resulting in irregular Poisson-like spike trains. Recent data show that the degree of irregularity in neuronal spike trains recorded during the delay period of working memory experiments is the same for both low-activity states of a few Hz and for elevated, persistent activity states of a few tens of Hz. Since the difference between these persistent activity states cannot be due to external factors coming from sensory inputs, this suggests that the underlying network dynamics might support coexisting balanced states at different firing rates. We use mean field techniques to study the possible existence of multiple balanced steady states in recurrent networks of current-based leaky integrate-and fire (LIF) neurons. To assess the degree of balance of a steady state, we extend existing mean-field theories so that not only the firing rate, but also the coefficient of variation of the interspike interval distribution of the neurons, are determined self-consistently. Depending on the connectivity parameters of the network, we find bistable solutions of different types. If the local recurrent connectivity is mainly excitatory, the two stable steady states differ mainly in the mean current to the neurons. In this case, the mean drive in the elevated persistent activity state is suprathreshold and typically characterized by low spiking irregularity. If the local recurrent excitatory and inhibitory drives are both large and nearly balanced, or even dominated by inhibition, two stable states coexist, both with subthreshold current drive. In this case, the spiking variability in both the resting state and the mnemonic persistent state is large, but the balance condition implies parameter fine-tuning. Since the degree of required fine-tuning increases with network size and, on the other hand, the size of the fluctuations in the afferent current to the cells increases for small networks, overall we find that fluctuation-driven persistent activity in the very simplified type of models we analyze is not a robust phenomenon. Possible implications of considering more realistic models are discussed. PMID- 17134317 TI - Exact subthreshold integration with continuous spike times in discrete-time neural network simulations. AB - Very large networks of spiking neurons can be simulated efficiently in parallel under the constraint that spike times are bound to an equidistant time grid. Within this scheme, the subthreshold dynamics of a wide class of integrate-and fire-type neuron models can be integrated exactly from one grid point to the next. However, the loss in accuracy caused by restricting spike times to the grid can have undesirable consequences, which has led to interest in interpolating spike times between the grid points to retrieve an adequate representation of network dynamics. We demonstrate that the exact integration scheme can be combined naturally with off-grid spike events found by interpolation. We show that by exploiting the existence of a minimal synaptic propagation delay, the need for a central event queue is removed, so that the precision of event-driven simulation on the level of single neurons is combined with the efficiency of time driven global scheduling. Further, for neuron models with linear subthreshold dynamics, even local event queuing can be avoided, resulting in much greater efficiency on the single-neuron level. These ideas are exemplified by two implementations of a widely used neuron model. We present a measure for the efficiency of network simulations in terms of their integration error and show that for a wide range of input spike rates, the novel techniques we present are both more accurate and faster than standard techniques. PMID- 17134318 TI - The road to chaos by time-asymmetric Hebbian learning in recurrent neural networks. AB - This letter aims at studying the impact of iterative Hebbian learning algorithms on the recurrent neural network's underlying dynamics. First, an iterative supervised learning algorithm is discussed. An essential improvement of this algorithm consists of indexing the attractor information items by means of external stimuli rather than by using only initial conditions, as Hopfield originally proposed. Modifying the stimuli mainly results in a change of the entire internal dynamics, leading to an enlargement of the set of attractors and potential memory bags. The impact of the learning on the network's dynamics is the following: the more information to be stored as limit cycle attractors of the neural network, the more chaos prevails as the background dynamical regime of the network. In fact, the background chaos spreads widely and adopts a very unstructured shape similar to white noise. Next, we introduce a new form of supervised learning that is more plausible from a biological point of view: the network has to learn to react to an external stimulus by cycling through a sequence that is no longer specified a priori. Based on its spontaneous dynamics, the network decides "on its own" the dynamical patterns to be associated with the stimuli. Compared with classical supervised learning, huge enhancements in storing capacity and computational cost have been observed. Moreover, this new form of supervised learning, by being more "respectful" of the network intrinsic dynamics, maintains much more structure in the obtained chaos. It is still possible to observe the traces of the learned attractors in the chaotic regime. This complex but still very informative regime is referred to as "frustrated chaos." PMID- 17134319 TI - Analysis and design of echo state networks. AB - The design of echo state network (ESN) parameters relies on the selection of the maximum eigenvalue of the linearized system around zero (spectral radius). However, this procedure does not quantify in a systematic manner the performance of the ESN in terms of approximation error. This article presents a functional space approximation framework to better understand the operation of ESNs and proposes an information-theoretic metric, the average entropy of echo states, to assess the richness of the ESN dynamics. Furthermore, it provides an interpretation of the ESN dynamics rooted in system theory as families of coupled linearized systems whose poles move according to the input signal dynamics. With this interpretation, a design methodology for functional approximation is put forward where ESNs are designed with uniform pole distributions covering the frequency spectrum to abide by the richness metric, irrespective of the spectral radius. A single bias parameter at the ESN input, adapted with the modeling error, configures the ESN spectral radius to the input-output joint space. Function approximation examples compare the proposed design methodology versus the conventional design. PMID- 17134320 TI - Invariant global motion recognition in the dorsal visual system: a unifying theory. AB - The motion of an object (such as a wheel rotating) is seen as consistent independent of its position and size on the retina. Neurons in higher cortical visual areas respond to these global motion stimuli invariantly, but neurons in early cortical areas with small receptive fields cannot represent this motion, not only because of the aperture problem but also because they do not have invariant representations. In a unifying hypothesis with the design of the ventral cortical visual system, we propose that the dorsal visual system uses a hierarchical feedforward network architecture (V1, V2, MT, MSTd, parietal cortex) with training of the connections with a short-term memory trace associative synaptic modification rule to capture what is invariant at each stage. Simulations show that the proposal is computationally feasible, in that invariant representations of the motion flow fields produced by objects self-organize in the later layers of the architecture. The model produces invariant representations of the motion flow fields produced by global in-plane motion of an object, in-plane rotational motion, looming versus receding of the object, and object-based rotation about a principal axis. Thus, the dorsal and ventral visual systems may share some similar computational principles. PMID- 17134321 TI - Recurrent cerebellar loops simplify adaptive control of redundant and nonlinear motor systems. AB - We have described elsewhere an adaptive filter model of cerebellar learning in which the cerebellar microcircuit acts to decorrelate motor commands from their sensory consequences (Dean, Porrill, & Stone, 2002). Learning stability required the cerebellar microcircuit to be embedded in a recurrent loop, and this has been shown to lead to a simple and modular adaptive control architecture when applied to the linearized 3D vestibular ocular reflex (Porrill, Dean, & Stone, 2004). Here we investigate the properties of recurrent loop connectivity in the case of redundant and nonlinear motor systems and illustrate them using the example of kinematic control of a simulated two-joint robot arm. We demonstrate that (1) the learning rule does not require unavailable motor error signals or complex neural reference structures to estimate such signals (i.e., it solves the motor error problem) and (2) control of redundant systems is not subject to the nonconvexity problem in which incorrect average motor commands are learned for end-effector positions that can be accessed in more than one arm configuration. These properties suggest a central functional role for the closed cerebellar loops, which have been shown to be ubiquitous in motor systems (e.g., Kelly & Strick, 2003). PMID- 17134322 TI - Free-lunch learning: modeling spontaneous recovery of memory. AB - After a language has been learned and then forgotten, relearning some words appears to facilitate spontaneous recovery of other words. More generally, relearning partially forgotten associations induces recovery of other associations in humans, an effect we call free-lunch learning (FLL). Using neural network models, we prove that FLL is a necessary consequence of storing associations as distributed representations. Specifically, we prove that (1) FLL becomes increasingly likely as the number of synapses (connection weights) increases, suggesting that FLL contributes to memory in neurophysiological systems, and (2) the magnitude of FLL is greatest if inactive synapses are removed, suggesting a computational role for synaptic pruning in physiological systems. We also demonstrate that FLL is different from generalization effects conventionally associated with neural network models. As FLL is a generic property of distributed representations, it may constitute an important factor in human memory. PMID- 17134323 TI - Linear multilayer ICA generating hierarchical edge detectors. AB - In this letter, a new ICA algorithm, linear multilayer ICA (LMICA), is proposed. There are two phases in each layer of LMICA. One is the mapping phase, where a two-dimensional mapping is formed by moving more highly correlated (nonindependent) signals closer with the stochastic multidimensional scaling network. Another is the local-ICA phase, where each neighbor (namely, highly correlated) pair of signals in the mapping is separated by MaxKurt algorithm. Because in LMICA only a small number of highly correlated pairs have to be separated, it can extract edge detectors efficiently from natural scenes. We conducted numerical experiments and verified that LMICA generates hierarchical edge detectors from large-size natural scenes. PMID- 17134324 TI - Functional network topology learning and sensitivity analysis based on ANOVA decomposition. AB - A new methodology for learning the topology of a functional network from data, based on the ANOVA decomposition technique, is presented. The method determines sensitivity (importance) indices that allow a decision to be made as to which set of interactions among variables is relevant and which is irrelevant to the problem under study. This immediately suggests the network topology to be used in a given problem. Moreover, local sensitivities to small changes in the data can be easily calculated. In this way, the dual optimization problem gives the local sensitivities. The methods are illustrated by their application to artificial and real examples. PMID- 17134325 TI - Second-order cone programming formulations for robust multiclass classification. AB - Multiclass classification is an important and ongoing research subject in machine learning. Current support vector methods for multiclass classification implicitly assume that the parameters in the optimization problems are known exactly. However, in practice, the parameters have perturbations since they are estimated from the training data, which are usually subject to measurement noise. In this article, we propose linear and nonlinear robust formulations for multiclass classification based on the M-SVM method. The preliminary numerical experiments confirm the robustness of the proposed method. PMID- 17134326 TI - Fast generalized cross-validation algorithm for sparse model learning. AB - We propose a fast, incremental algorithm for designing linear regression models. The proposed algorithm generates a sparse model by optimizing multiple smoothing parameters using the generalized cross-validation approach. The performances on synthetic and real-world data sets are compared with other incremental algorithms such as Tipping and Faul's fast relevance vector machine, Chen et al.'s orthogonal least squares, and Orr's regularized forward selection. The results demonstrate that the proposed algorithm is competitive. PMID- 17134328 TI - Advancing personality assessment terminology: time to retire "objective" and "projective" as personality test descriptors. PMID- 17134330 TI - Phenomenology, Bruno Klopfer, and individualized/collaborative assessment. AB - This presentation celebrates commonalities of Bruno Klopfer's philosophical phenomenological approach to the Rorschach and my similarly grounded approach to individualized/collaborative psychological assessment. The article begins with an excerpt from such an assessment to ground what follows. It then addresses what phenomenology is and is not in relation to psychological assessment, and reviews Klopfer's phenomenological approach to the Rorschach. It presents the findings from an empirical phenomenological research study relevant to collaborative assessment. Finally, the article reviews some common inquiries and observations about individualized/colloaborative assessment and closes with some shifts in American psychology that are consonant with this approach to psychological assessment. PMID- 17134331 TI - Computers, criminals, an eccentric billionaire, and APA: a brief autobiography. AB - In this invited autobiographical account, I sum up what life has been like for me personally and professionally. For most of the first 50 years of my life, I lived in Alabama. During my years at the University of Alabama, my professional activities included developing a computer-based system to interpret the MMPI (Hathaway & McKinley, 1943), managing a national and international continuing education program for psychologists, involvement in a class action suit that resulted in the deinstitutionalization of Alabama's mental hospitals, organizing a team of professionals to reclassify all of the inmates of Alabama's prison system, and conducting a psychological autopsy on Howard Hughes. I was the American Psychological Association (APA) president in 1988 and served from 1989 to 2003 as APA Chief Executive Officer. Since my time at APA, I have been engaged in work with international psychological organizations. PMID- 17134332 TI - Assessing cross-cultural differences through use of multiple-group invariance analyses. AB - The use of structural equation modeling in cross-cultural personality research has become a popular method for testing measurement invariance. In this report, we present an example of testing measurement invariance using the Sense of Coherence Scale of Antonovsky (1993) in 3 ethnic groups: Chinese, Japanese, and Whites. In a series of increasingly restrictive constraints on the measurement models of the 3 groups, we demonstrate how to assess differences among the groups. We also provide an example of construct validation. PMID- 17134333 TI - Sample size in clinical research: when is enough enough? PMID- 17134334 TI - Incremental validity of the typical intellectual engagement scale as predictor of different academic performance measures. AB - The incremental validity of the Typical Intellectual Engagement (TIE) scale (Goff & Ackerman, 1992) as a predictor of academic performance (AP) was tested over and above other established determinants of AP, namely, psychometric g (as extracted from 5 cognitive ability tests) and the Big Five personality traits, assessed by the Neuroticism-Extraversion-Openness Five Factor Inventory (Costa & McCrae, 1992). One hundred four British students were tested on arrival to university, and AP measures were collected longitudinally throughout a 3-year period. TIE, g, and Conscientiousness were the highest correlates of AP. A series of multiple hierarchical regressions showed that TIE had significant incremental validity (over and above g and the Big Five) in the prediction of AP. Implications are discussed in light of the investment theory of intellectual competence and the utility of self-report inventories as predictors of academic achievement. PMID- 17134335 TI - Socially desirable responding on the web: investigating the candor hypothesis. AB - The investigation presented here explores the hypothesis that participants are less likely to respond in a socially desirable fashion on self-report questionnaires completed on the Web relative to those completed in the laboratory -the candor hypothesis. A battery of social desirability questionnaires (i.e., Balanced Inventory of Desirable Responding [Paulhaus, 1984], Marlowe-Crowne Social Desirability Scale [Crowne & Marlowe, 1964], Eysenck Personality Questionnaire-Revised Lie Scale [Eysenck & Eysenck, 1994]) was administered to 3 groups: 2 groups consisted of undergraduate participants who were randomly assigned to complete the measures either in the laboratory (n = 60) or on the Web (n = 60), and 1 group consisted of self-selected participants who visited our experimental Web page and completed the measures online (n = 284). This design allowed us to assess the role of Web administration while controlling for differences in sample type, an oft-neglected issue in the Web literature. Results do not support the claim that administering self-report measures over the Web results in a decrease in socially desirable responding. Furthermore, these findings highlight the problems associated with confounding sample and medium. Implications for the use of Web as a research tool are discussed. PMID- 17134336 TI - The thematic apperception test: toward a standard measure of the big three motives. AB - The application of the Thematic Apperception Test to the assessment of motives has been heralded as an important milestone in personality psychology. However, although this approach is well established, there is at present no standard battery of cues for measuring the Big Three motives (achievement, affiliation, power). Furthermore, the extent to which scoring subcategories contribute to overall motive scores has been neglected. Our research with students and managers examined the effectiveness of picture cues in eliciting motive imagery and the prevalence of scoring subcategories within each motive scoring system. Results from 2 data sets comprising 547 men and women suggested that there were 3 cues that should be retained for future research and that motive scoring systems could be refined through removal of redundant subcategories. Further research is needed to systematically investigate the effectiveness of a standard battery of cues and the validity of revised motive scoring systems. PMID- 17134337 TI - Confirmatory factor analysis of the three-factor structure of the schizotypal personality questionnaire and Chapman schizotypy scales. AB - We examined the factor structure of the Schizotypal Personality Questionnaire (SPQ; Raine, 1991), using confirmatory factor analysis in 3 experiments, with an aim to better understand the construct of schizotypy. In Experiment 1 we tested the fit of 2-, 3-, and 4-factor models on SPQ data from a normal sample. The paranoid 4-factor model fit the data best but not adequately. Based on the strong basis for the Raine 3-factor model we attempted to improve the fit of the 3 factor model by making 3 modifications to the Raine model. These modifications produced a well-fitting model. In Experiment 2 the good fit of this modified 2 factor model to SPQ scores was replicated in an independent normal sample. In Experiment 3, the modified 3-factor model was successfully extended to include the 3 Chapman schizotypy scales. Together these 3 experiments indicate that the 3 factor model of the SPQ, albeit with some slight modifications, is a good model for schizotypy structure that is not restricted to 1 measure of schizotypal personality traits. PMID- 17134338 TI - Curiosity about people: the development of a social curiosity measure in adults. AB - Curiosity refers to the desire for acquiring new information. The aim of this study was to develop a questionnaire to assess social curiosity, that is, interest in how other people think, feel, and behave. The questionnaire was administered to 312 participants. Factor analyses of the 10-item Social Curiosity Scale (SCS) yielded 2 factors: General Social Curiosity and Covert Social Curiosity. Evidence of convergent validity was provided by moderately high correlations of the SCS with other measures of curiosity and self-perceived curiosity, whereas discriminant validity was demonstrated by low correlations of the SCS with other personality traits, such as neuroticism and agreeableness. Of interest, social interaction anxiety was observed to facilitate covert social curiosity while inhibiting general social curiosity. PMID- 17134339 TI - Interpersonal traits in childhood: development of the child and adolescent interpersonal survey. AB - The Child and Adolescent Interpersonal Survey (CAIS) consists of interpersonal trait descriptions that were generated to represent the constructs of the Interpersonal Circumplex model utilizing language accessible to children in a brief self-report format. Scale development entailed examining the structure at the item and scale levels with a sample of children (fourth and sixth graders) and at the scale level with college students. Internal consistency estimates for the majority of the CAIS scales were adequate given the brevity of the scales. The fits to the circular order model for the CAIS were very well supported across the child and adult samples, with no significant differences in the fit of the model across these samples. Generally strong associations were found between corresponding scales of the CAIS and the Interpersonal Adjective Scales in a college sample, and there were many structural similarities between these measures. For the child sample, the CAIS scales of Gregarious-Extraverted and Warm-Agreeable related strongly to corresponding Five-factor scales, and remaining scales related minimally to moderately. PMID- 17134340 TI - A Rorschach stability study in a nonpatient adult sample. AB - The objective of this study was to provide new primary data on Rorschach Comprehensive System stability levels. To achieve this, we tested 75 French nonpatient adults twice on the Rorschach with a 3-month interval between the tests. Interrater reliability was in the excellent range for most of the variables studied. The overall stability level in a selected set of previously studied variables was below expectations (median r = .53). Personality, cognitive or self/relational variables yielded higher test-retest correlations than emotional and coping variables. Moderators of stability could be identified: (a) overall level of Task Engagement (TE) in F, m, FM + m, a, FC, Sum C', Sum V, Sum Shd, Fr + rF, INC + FAB, COP, es, Adj es, EGO, and Blends; (b) variations in TE in F, FM, and p; (c) state distress in Zd, m, FM + m, a, C, CF + C, WSumC, FD, and es; (d) variables derived from the number of responses impacted stability in P, Zf, m, FC, CF + C, Sum C', Sum V, MOR, EA, es, and Blends. These results provide further support for the reliability of several measures. Examiner effects as an influence on productivity and TE were identified as an important area for future research. PMID- 17134344 TI - Standards for education and training in psychological assessment position of the Society for Personality Assessment. AB - The Society for Personality Assessment is a national and international professional organization devoted to research and practice in the field of psychological assessment. As such, it represents practitioners of assessment regardless of discipline or degree. PMID- 17134347 TI - The experts speak. New issues in the challenge of HIV drug resistance. Interview by Vicki Glaser. PMID- 17134348 TI - Three-class antiretroviral resistance in a patient with acute HIV-1 infection. AB - Antiretroviral resistance in treatment-naive patients with HIV-1 infection is on the rise in both resource-rich and resource-poor countries. We report a case of three-class primary antiretroviral resistance detected by a genotypic assay in a 19-year-old woman with acute HIV-1 infection. Her risk factor for HIV acquisition was unprotected sexual intercourse with a HIV-positive man. At diagnosis, her HIV 1 RNA level was 730,376 copies per milliliter, and 2 weeks later her CD4+ cell count was 465 cells/mm(3). The patient's antiretroviral therapy was chosen based on the genotype of the source patient. Subsequent analysis of our patient's virus revealed an identical genotype to that of the source patient. The patient tolerated therapy well and continues to be virologically suppressed after 1 year of therapy. Her current CD4+ cell count is 537 cells/mm(3) and HIV-1 RNA is less than 400 copies per milliliter. This case supports the current recommendations from several HIV therapy guidelines to perform resistance testing in patients with acute HIV-1 infection. PMID- 17134349 TI - Successful treatment with atazanavir and lopinavir/ritonavir combination therapy in protease inhibitor-susceptible and protease inhibitor-resistant HIV-infected patients. AB - The combination of atazanavir (ATV) plus lopinavir/ritonavir (LPV/r) has been used in practice. However, clinical data supporting its use are limited. The objective of this study was to evaluate the efficacy and tolerability of regimens with ATV + LPV/r in protease inhibitor (PI)-susceptible and PI-resistant patients. A retrospective review of 2703 charts was performed to identify all patients who received ATV + LPV/r. From June 2003 to January 2005, 33 patients received ATV + LPV/r with nucleoside reverse transcriptase inhibitors (NRTIs) for 3 months or more. Virologic success (HIV-RNA < 400 copies per milliliter) was achieved in 30 patients (91%) in a median of 10 weeks (range, 2-68). Nineteen of the 23 patients (83%) who had ultrasensitive viral load (VL) assays were nondetectable. Among patients with 6 or more protease resistance (PR) mutations (PI-resistant), 11 of 14 (79%) achieved virologic success. Eleven of those received phenotypic testing (10 Virtual Phenotype, VircoLab, Baltimore, MD). Despite predicted phenotypic resistance to ATV (6 patients) and LPV/r (7 patients), virologic success was achieved in 4 of 6 (67%) and 4 of 7 (57%), respectively. The 3 PI-resistant patients who were virologic failures had extensive prior LPV/r use, 8-11 PR mutations, and predicted phenotypic resistance to LPV/r, but 2 of 3 had CD4 increases with ATV + LPV/r. Overall, 28 patients (85%) continue to tolerate ATV + LPV/r for a median of 32 weeks follow-up (range, 12-76). Combination ATV + LPV/r with NRTIs appears safe, tolerable, and efficacious in PI-resistant patients (>/=6 PR mutations) and predicted phenotypic resistance to ATV and LPV/r. Further studies of ATV + LPV/r in HIV-treatment are warranted. PMID- 17134350 TI - A comprehensive evaluation of survey questions for adherence to antiretroviral medications and exploratory analyses for identifying optimal sets of survey questions. AB - Although many methods for assessing adherence have been developed, most are not feasible for busy clinical settings. Using patients from the Adherence and Efficacy of Protease inhibitor Therapy (ADEPT) study (1998-2000), we systematically evaluated the relationship between psychosocial, environmental, clinical, and other factors with adherence to create composite variables (CVs) that are efficient with high sensitivity for detecting nonadherence and great potential for busy clinics. Eligible patients were protease inhibitor naive or started a regimen within 3 months from baseline. Of the 128 patients who responded to survey at baseline, weeks 8, 24, and/or 48, mean (standard deviation [SD]) age was 39.3 (8.2) years with 81% male. About half of the patients were Latino, followed by 28% African American and 14% Caucasian. Sixteen percent reported injection drug use, and 40% had male-male sex. Mean CD4 count was 184.8 cells/mm(3) with a range from 1 to 1130 cells/mm(3). Thirty-two variables had a significant association with adherence at one or two time points and 9 were significantly associated with adherence over time. Among these significant factors, 8 also had a relationship with a clearly monotonic trend, by which 219 CVs were formed. Among these CVs, 8 were significantly associated with adherence and had a relationship with monotonic trend. Compared to traditional self reported adherence, CVs had much higher sensitivities (p < 0.001) for detecting nonadherence. We conclude that CVs consisting of a combination of psychological, behavior, and adherence questions may be reasonable substitutes for direct adherence questions, which are limited by problems with recall and social biases. Trust in physicians, having a child, history of substance use, CD4 count, and belief that antiretrovirals can help living longer or improve quality of life can efficiently predict nonadherence. Because these variables are readily obtainable in clinical settings, these selected questions may provide a clinically useful means of screening patients for antiretroviral medication nonadherence. PMID- 17134351 TI - A randomized trial of the impact of a programmable medication reminder device on quality of life in patients with AIDS. AB - This 6-month randomized controlled trial evaluated the impact on quality of life (QOL) of a medication reminder device for patients with HIV. Patients were eligible if they had taken three or fewer highly active antiretroviral therapy (HAART) regimens or were treatment naive. The intervention group received the Disease Management Assistance System (DMAS), a prompting device that verbally reminds patients at medication times and electronically records doses, and a monthly 30 minute adherence educational session. Controls received education only. QOL was measured at baseline and 6 months using the Centers for Epidemiologic Studies Depression Scale (CES-D), Instrumental Activities of Daily Living (IADLs), and the Medical Outcomes Study HIV Health Survey (MOS-HIV). At baseline, 62 patients completed surveys (31 control, 31 DMAS); at month 6, 48 patients completed surveys (23 control, 25 DMAS). At month 6, controls had improved QOL scores for CES-D, IADLs, physical health, general health, pain, QOL, and role functioning, while participants in the DMAS arm had some deterioration in QOL scores. These differences persisted after controlling for demographics, baseline CD4, and adherence. DMAS was associated with improved adherence but decreased QOL. PMID- 17134352 TI - Association of comorbidity with physical disability in older HIV-infected adults. AB - Comorbidity, aging, and their impact on physical functioning will play an increasingly greater role in HIV medical care as the number of infected adults over 50 years of age grows. The study objective was to investigate the relationship of comorbidity and age with physical functioning in HIV-infected and HIV-negative patients. Eight hundred eighty-nine HIV-infected veterans and 647 HIV-negative veterans from the Veterans Aging Cohort Study conducted between September 2001 and June 2002 were included in the study. Physical functioning was measured by self-reported difficulty with various physical activities. Regression analyses were performed to examine demographic and clinical factors associated with physical functioning. Separate models were used for HIV-infected and HIV negative subjects since these groups differed in demographic makeup. In both patient groups, chronic lung disease, coronary artery disease, hypertension, smoking, and major depression were independently associated with reduced physical functioning in age and race adjusted regression models. Increased age was associated with reduced physical functioning in both HIV-infected and HIV negative patients. However, when comorbid conditions were entered into the models for both HIV-infected and HIV-negative patients, age coefficients were reduced and were no longer statistically significant. Among the HIV-infected patients, results remained unchanged after controlling for the impact of antiretroviral therapy and HIV disease stage. Our findings demonstrate the important role of general medical comorbidity in physical functioning in both HIV-infected and HIV negative patients. This suggests the importance of effectively treating comorbid conditions in persons with HIV, in order to reduce the overall impact of disease on physical functioning. PMID- 17134353 TI - The intent and practice of condom use among HIV-positive men who have sex with men in Japan. AB - To evaluate the intent and practice of condom use among Japanese HIV-positive men who have sex with men (MSM), a survey using anonymous questionnaires was carried out and 117 respondents were investigated. For anal sex and oral sex, respectively, 58.1% and 15.2% intended to use condoms and 47.2% and 12.4% used condoms all of the time. The intent of condom use decisively affected the practice of condom use and was closely related to the perceived risk level of HIV/sexually transmitted (STI) transmission. In anal sex, willingness to protect sexual partners from HIV infection was strongly related not only to the intent but also to the practice. Enhancement of willingness to protect oneself from STI was suggested to enhance willingness to protect his/her sexual partners from HIV infection with secondary enhancement of the intent or the practice of condom use. Specific support of MSM with HIV for improving the intent and practice of condom use is urgently needed. PMID- 17134354 TI - HIV-related knowledge, attitudes, perceived benefits, and risks of HIV testing among pregnant women in rural Southern India. AB - The rising prevalence of HIV among pregnant women in rural India is of great concern. Prenatal voluntary counseling and HIV testing (VCT) is critical to prevent mother-to-child transmission of HIV (PMTCT). We surveyed 202 pregnant women attending a rural antenatal clinic in Southern India to investigate HIV related knowledge, attitudes toward infant feeding practices, and perceived benefits and risks of HIV testing. Of the total of 202 women surveyed, 189 women (94%) had heard of HIV/AIDS and 60% of them had relatively good knowledge regarding risk factors for HIV transmission. However, 48% did not know that there are "means to prevent mother-to-child HIV transmission." If women were not to breastfeed her baby, negative attitudes expected from the partner would include 84% thinking that that the mother is harming the baby, 78% thinking she is not a good mother, 74% thinking she has HIV, and 66% thinking she has been unfaithful. Ninety-seven percent of women did not perceive themselves at risk for HIV and only 57% had been tested for HIV. Although, 85% of women expressed their willingness to be tested, most were concerned about confidentiality and disclosing HIV serostatus because of fear of negative reactions from their husbands, parents, and community. Many social and cultural barriers confront pregnant women when they decide to opt for HIV testing. If VCT and PMTCT interventions are to be successful, urgent attention must be focused on education, development of innovative culturally appropriate interventions that empower women to make decisions about HIV testing, involvement of men, and addressing stigma and discriminatory attitudes toward people living with HIV/AIDS. PMID- 17134355 TI - New atazanavir capsule approved. PMID- 17134356 TI - FDA approves early test. PMID- 17134357 TI - The cancer stem cell: a new therapeutic paradigm? PMID- 17134359 TI - Cetuximab in the treatment of head and neck cancer. AB - Approaches to the treatment of locally advanced and recurrent and/or metastatic squamous cell carcinoma of the head and neck (SCCHN) have been limited by their toxicity. Effective, better tolerated approaches are urgently required. Cetuximab is an immunoglobulin G(1) monoclonal antibody that specifically targets the epidermal growth factor receptor (EGFR), which is commonly expressed in a number of solid tumors, including SCCHN, where it is associated with poor prognosis. Cetuximab is approved in 56 countries for use in the treatment of EGFR-expressing metastatic colorectal cancer that has progressed on irinotecan-containing therapy and has recently received approval in Europe and the USA for use in the treatment of SCCHN. A randomized Phase III study has demonstrated that cetuximab plus radiotherapy can significantly improve locoregional control and prolong overall survival compared with radiotherapy alone. Cetuximab has also been confirmed to be effective as monotherapy in recurrent and/or metastatic SCCHN that has progressed on platinum-containing therapy. Clinical studies have demonstrated that cetuximab is well tolerated and does not significantly increase the side effects associated with radiotherapy or chemotherapy. This article presents the rationale for EGFR inhibition in the management of head and neck cancers, and the preclinical and clinical evidence for the use of cetuximab in the treatment of SCCHN. PMID- 17134360 TI - Hepatocellular cancer in HIV-infected individuals: tomorrow's problem? AB - A number of cancers occur with increased frequency in individuals with HIV infection. There is a debate as to whether or not hepatocellular cancer is one of these tumors. This article reviews the current data regarding incidence etiology, treatment and outcome for HIV-related hepatocellular cancer. It also attempts to predict future areas of importance in the management of these patients in the era of highly active antiretroviral therapy. PMID- 17134361 TI - Targeting events in melanoma carcinogenesis for the prevention of melanoma. AB - Melanoma is one of the few tumors that have increased in incidence over the last few decades. Strategies devoted solely to protecting against ultraviolet radiation have, at best, had a modest impact on the development of melanoma. Chemoprevention is an under-explored approach that could significantly decrease the morbidity and mortality from this deadly cancer. However, the scientific and logistical challenges of performing clinical studies in chemoprevention require innovative approaches to prove the effectiveness of putative preventive agents. There are several pharmacological and nutriceutical agents that are mechanistically linked to events in melanoma carcinogenesis that are candidates for advanced human studies. We will review the data for several promising agents, including statins, curcumin, resveratrol, silymarin and green tea, and discuss some importance issues and concepts that should be considered in any melanoma chemoprevention strategy. PMID- 17134362 TI - Current surgical management of melanoma. AB - The surgical management of melanoma has changed dramatically over the last few decades. Through the development and conduction of well-designed, prospective, randomized trials, we have been able to refine the way that we surgically manage patients with melanoma. Indeed, many important issues have been addressed through such trials: the proper surgical margins for the primary melanoma, utility of the elective lymph node dissection and the role for selective lymphadenectomy, to name a few. This review will also discuss what we have learned from past clinical trials and address several issues with regards to where we are going in the future. PMID- 17134363 TI - Oncolytic viruses and DNA-repair machinery: overcoming chemoresistance of gliomas. AB - The current standard of care for malignant gliomas is surgical resection and radiotherapy followed by extended adjuvant treatment with the alkylating agent temozolomide. Temozolomide causes DNA damage, which induces cell death. Through changes in the DNA-repair machinery, glioma cells develop resistance to temozolomide, compromising the therapeutic effect of the drug. Oncolytic viruses, such as herpes simplex viruses and adenoviruses, are being introduced into clinical trials as a new treatment for this malignancy. Biological studies have revealed that these viruses use mechanisms to either inactivate (adenovirus) or take advantage of (herpes simplex virus) the cellular DNA-repair machinery to achieve productive replication. Adenoviruses express proteins from the early genes to either downregulate the damage-repair enzyme, O(6)-methylguanine-DNA methyltransferase, or degrade poly (ADP-ribose) polymerase or the Mre11-Rad50 NBS1 complex, which detects DNA strand breaks. Temozolomide enhances herpes simplex virus oncolysis by upregulating the DNA repair-related genes growth arrest DNA damage 34 and ribonucleotide reductase. The interactions between viruses and the DNA-repair machinery suggest that a combined temozolomide and viral therapy will overcome the limitations of a single therapy by diminishing chemoresistance or enhancing oncolysis. This hypothesis has been supported by promising findings from preclinical and clinical studies. PMID- 17134364 TI - Recurrent glioblastoma multiforme: advances in treatment and promising drug candidates. AB - Recurrent glioblastoma multiforme is a lethal disease with currently available treatment options having a limited impact on outcome. In this article, current and novel therapeutic approaches in the treatment of recurrent glioblastoma multiforme, including chemotherapy, targeted molecular agents, virotherapy/gene therapy and immunotherapy and challenges in developing novel therapeutic agents for glioblastoma multiforme will be discussed. PMID- 17134365 TI - Primary gastrointestinal tract lymphoma: diagnosis and management of common neoplasms. AB - Primary gastrointestinal lymphoma represents the most common location of extranodal lymphoma. With the bulk of disease manifesting within the gastrointestinal tract and contiguous lymph nodes, many of the lymphomas occurring in the peripheral lymph nodes can also present with primary gastrointestinal tract involvement. Molecular biology has recently enabled significant progress in the diagnosis and management of primary gastrointestinal lymphoma. Herein, we will discuss the major lymphomas affecting the bowel and highlight their key morphological, immunophenotypical and molecular diagnostic attributes. Similarly, in keeping with recent therapeutic advances, we will briefly discuss some important treatment considerations. Thus, this review is intended to offer clinicians and pathologists an overview of primary gastrointestinal lymphomas. PMID- 17134366 TI - Sequelae following axillary lymph node dissection for breast cancer. AB - Axillary lymph node dissection (ALND) has a central role in the surgical management of breast cancer; however, it is associated with a potentially significant morbidity. Although post-ALND complications are often minor, in some cases they can persist for a long time following surgery, thereby affecting the quality of life of breast cancer survivors. Seroma formation and altered sensation of the upper limb are the two most common complications following ALND. Lymphedema is the most common potentially severe long-term complication following ALND. Major post-ALND complications (such as injury or thrombosis of the axillary vein and injury to the motor nerves of the axilla) are extremely rare. Meticulous surgical technique and careful selection of patients for postoperative radiation therapy are mandatory to prevent significant morbidity following ALND. The introduction of the technique of sentinel lymph node biopsy in clinical practice has resulted in a significant reduction in the incidence of post-ALND complications. PMID- 17134367 TI - Evolving treatment paradigms for locally advanced and metastatic prostate cancer. AB - While men with early stage prostate cancer typically enjoy long-term survival after definitive management, for those who present with locally advanced or metastatic disease, survival is compromised. Multimodality therapy can prolong survival in these patients, with state-of-the-art options including intensity modulated radiation or brachytherapy in conjunction with androgen ablation, adjuvant androgen ablation and/or chemotherapy with radical retropubic prostatectomy. In addition, novel biological therapies are being explored to target the unique molecular changes in prostate cancer cells and their interactions with the microenvironment. With these advances the outlook will undoubtedly improve, even for patients presenting with advanced disease. Careful application of these emerging therapies to a select group of prostate cancer patients most likely to obtain benefit from them is the challenge for urologists, medical oncologists and radiation oncologists for the future. PMID- 17134368 TI - Effects of adjuvant aromatase inhibitor therapy on lipid profiles. AB - How aromatase inhibitors affect lipids is of great interest. Compared with tamoxifen, adjuvant anastrozole and letrozole are associated with increased incidences of hypercholesterolemia, while similar data are lacking for exemestane in the adjuvant setting. No significant differences in lipid profiles occurred with extended adjuvant exemestane compared with placebo, but total cholesterol and low-density lipoprotein levels increased significantly above baseline in both groups over 6 months. Likewise, no significant differences in hypercholesterolemia rates occurred between extended adjuvant letrozole and placebo. A lipid substudy further confirmed that letrozole did not significantly alter serum lipids for 36 months compared with placebo. Thus, although aromatase inhibitors lack the lipid-lowering properties of tamoxifen, no significant worsening of lipid levels occurs with their use. Patients would benefit from lifestyle changes and routine monitoring of serum lipids. Breast cancer therapy trials often report serum lipid parameters, but assessing the quality and overall significance of the data can be difficult. Methodology of data collection varies among trials and the concomitant use of lipid-modifying medication is often not reported. This review discusses the current understanding of the influence of lipid levels on cardiovascular risk in women and presents key findings on the effects of adjuvant aromatase inhibitor therapy on lipid profiles. PMID- 17134369 TI - Perspectives of proteomics in acute myeloid leukemia. AB - Acute myeloid leukemia (AML) is a frequent hematological malignancy. Despite enormous therapeutic efforts that range from various cytotoxic agents to allogeneic stem cell transplantation, overall survival of patients with AML remains unsatisfying. The poor survival rates are mainly due to therapy-related mortality, failure of induction chemotherapy and early relapses. Therefore, novel therapeutic agents that are more efficient and better tolerated are eagerly sought after. For existing therapeutic strategies, there is a lack of markers that are capable of reliably predicting prognosis or the therapeutic response prior to treatment. There is hope that elucidation of the AML-specific proteome will prompt the discovery of novel therapeutic targets and biomarkers in AML. Modern mass-spectrometry instrumentation has achieved excellent performance in terms of sensitivity, resolution and mass accuracy; however, so far, the contribution of proteomics to the care of patients with AML is virtually zero. This might be partly because mass spectrometry instrumentation and protein fractionation still lack true high-throughput capabilities with highest levels of reproducibility, thus hampering large-scale translational studies with clinical samples. Since mass-spectrometry instruments are very intricate devices, their successful operation will hinge on the willingness and ability of mass spectrometry experts and clinical researchers to adopt new views, learn from each other and cooperate in order to ultimately benefit the patient suffering from AML. This review highlights some clinical problems circumventing the treatment of patients with AML. Furthermore, it provides a brief overview of the technical background of standard proteomics approaches and describes opportunities, challenges and pitfalls of proteomic studies with regards to AML. PMID- 17134370 TI - hMaxi-K gene transfer in males with erectile dysfunction: results of the first human trial. AB - Eleven patients with moderate to severe erectile dysfunction (ED) were given a single-dose corpus cavernosum injection of hMaxi-K, a "naked" DNA plasmid carrying the human cDNA encoding hSlo (for human slow-poke), the gene for the alpha, or pore-forming, subunit of the human smooth muscle Maxi-K channel. Three patients each were given 500, 1000, and 5000 microg, and two patients were given 7500 microg, of hMaxi-K and monitored for 24 weeks. The primary objectives of this phase I study were safety and tolerability of escalating hMaxi-K doses as assessed by clinical evaluations and laboratory tests. Secondary efficacy objectives were measured primarily by use of the International Index of Erectile Function (IIEF) scale. Patient responses were validated by partner responses. There were no serious adverse events and no dose-related adverse events attributed to gene transfer for any patient at any dose or study visit. No clinically significant changes from baseline were seen in physical evaluations (general and genitourinary), hematology, chemistry, and hormone analyses, or in cardiac events evaluated by repeated electrocardiograms. Importantly, no plasmid was detected in the semen of patients at any time after the injections. Patients given the two highest doses of hMaxi-K had apparent sustained improvements in erectile function (EF) as indicated by improved IIEF-EF domain scores over the length of the study. One patient given 5000 microg and one given 7500 microg reported EF category improvements that were highly clinically significant and were also maintained through the 24 weeks of study. Efficacy conclusions cannot be drawn from results of a phase I trial with no control group. However, the promising primary safety outcomes of the study and preliminary indications of effectiveness provide evidence that hMaxi-K gene transfer is a viable approach to the treatment of ED and that further studies investigating the efficacy of hMaxi K in patients with ED should be performed. PMID- 17134372 TI - Measurement of single-molecule conductance. AB - What is the conductance of a single molecule? This basic and seemingly simple question has been a difficult one to answer for both experimentalists and theorists. To determine the conductance of a molecule, one must wire the molecule reliably to at least two electrodes. The conductance of the molecule thus depends not only on the intrinsic properties of the molecule, but also on the electrode materials. Furthermore, the conductance is sensitive to the atomic-level details of the molecule-electrode contact and the local environment of the molecule. Creating identical contact geometries has been a challenging experimental problem, and the lack of atomic-level structural information of the contacts makes it hard to compare calculations with measurements. Despite the difficulties, researchers have made substantial advances in recent years. This review provides an overview of the experimental advances, discusses the advantages and drawbacks of different techniques, and explores remaining issues. PMID- 17134371 TI - Immunosuppressive strategies that are mediated by tumor cells. AB - Despite major advances in understanding the mechanisms leading to tumor immunity, a number of obstacles hinder the successful translation of mechanistic insights into effective tumor immunotherapy. Such obstacles include the ability of tumors to foster a tolerant microenvironment and the activation of a plethora of immunosuppressive mechanisms, which may act in concert to counteract effective immune responses. Here we discuss different strategies employed by tumors to thwart immune responses, including tumor-induced impairment of antigen presentation, the activation of negative costimulatory signals, and the elaboration of immunosuppressive factors. In addition, we underscore the influence of regulatory cell populations that may contribute to this immunosuppressive network; these include regulatory T cells, natural killer T cells, and distinct subsets of immature and mature dendritic cells. The current wealth of preclinical information promises a future scenario in which the synchronized blockade of immunosuppressive mechanisms may be effective in combination with other conventional strategies to overcome immunological tolerance and promote tumor regression. PMID- 17134373 TI - Fetal gene transfer using lentiviral vectors: in vivo detection of gene expression by microPET and optical imaging in fetal and infant monkeys. AB - Fetal intraperitoneal administration of human immunodeficiency virus (HIV)-l derived lentiviral vectors (10(7) infectious particles/fetus) has consistently shown high levels of transduction and gene expression in the omentum, peritoneum, and diaphragm when assessed by polymerase chain reaction (PCR) and whole tissue fluorescence. In vivo imaging techniques were explored with early-gestation long tailed macaques that were administered the vesicular stomatitis virus glycoprotein (VSV-G)-pseudotyped HIV-1-derived lentiviral vector expressing a mutant herpes simplex virus type 1 thymidine kinase (HSV-1-sr39tk) and firefly luciferase under the control of the cytomegalovirus (CMV) promoter. Fetuses were monitored sonographically and twice during gestation 9-[4-[18F]Fluoro-3 (hydroxymethyl)butyl]guanine (18F-FHBG) was injected into the fetal circulation under ultrasound guidance in preparation for microPET imaging. All newborns were delivered at term by cesarean section and raised in the nursery for postnatal studies. At 2 months postnatal age, animals were imaged and biodistribution was assessed. Optical imaging for firefly luciferase expression was also performed every 2 months postnatal age. Under all imaging conditions gene expression was observed in the abdominal region, and closely paralleled findings from prior studies based on whole tissue fluorescence. These investigations have shown that HSV-1-sr39tk and firefly luciferase can be used to safely detect transgene expression at multiple time points in fetal and infant monkeys in vivo and without evidence of adverse effects. PMID- 17134374 TI - Simultaneous determination of N7-alkylguanines in DNA by isotope-dilution LC tandem MS coupled with automated solid-phase extraction and its application to a small fish model. AB - In the present study, we report the development of a sensitive and selective assay based on LC (liquid chromatography)-MS/MS (tandem MS) to simultaneously measure N7-MeG (N7-methylguanine) and N7-EtG (N7-ethylguanine) in DNA hydrolysates. With the use of isotope internal standards (15N5-N7-MeG and 15N5-N7 EtG) and on-line SPE (solid-phase extraction), the detection limit of this method was estimated as 0.42 fmol and 0.17 fmol for N7-MeG and N7-EtG respectively. The high sensitivity achieved here makes this method applicable to small experimental animals. This method was applied to measure N7-alkylguanines in liver DNA from mosquito fish (Gambusia affinis) that were exposed to NDMA (N nitrosodimethylamine) and NDEA (N-nitrosodiethylamine) alone or their combination over a wide range of concentrations (1-100 mg/l). Results showed that the background level of N7-MeG in liver of control fish was 7.89+/-1.38 mmol/mol of guanine, while N7-EtG was detectable in most of the control fish with a range of 0.05-0.19 mmol/mol of guanine. N7-MeG and N7-EtG were significantly induced by NDMA and NDEA respectively, at a concentration as low as 1 mg/l and increased in a dose-dependent manner. Taken together, this LC-MS/MS assay provides the sensitivity and high throughput required to evaluate the extent of alkylated DNA lesions in small animal models of cancer induced by alkylating agents. PMID- 17134375 TI - Different redox states of metallothionein/thionein in biological tissue. AB - Mammalian metallothioneins are redox-active metalloproteins. In the case of zinc metallothioneins, the redox activity resides in the cysteine sulfur ligands of zinc. Oxidation releases zinc, whereas reduction re-generates zinc-binding capacity. Attempts to demonstrate the presence of the apoprotein (thionein) and the oxidized protein (thionin) in tissues posed tremendous analytical challenges. One emerging strategy is differential chemical modification of cysteine residues in the protein. Chemical modification distinguishes three states of the cysteine ligands (reduced, oxidized and metal-bound) based on (i) quenched reactivity of the thiolates when bound to metal ions and restoration of thiol reactivity in the presence of metal-ion-chelating agents, and (ii) modification of free thiols with alkylating agents and subsequent reduction of disulfides to yield reactive thiols. Under normal physiological conditions, metallothionein exists in three states in rat liver and in cell lines. Ras-mediated oncogenic transformation of normal HOSE (human ovarian surface epithelial) cells induces oxidative stress and increases the amount of thionin and the availability of cellular zinc. These experiments support the notion that metallothionein is a dynamic protein in terms of its redox state and metal content and functions at a juncture of redox and zinc metabolism. Thus redox control of zinc availability from this protein establishes multiple methods of zinc-dependent cellular regulation, while the presence of both oxidized and reduced states of the apoprotein suggest that they serve as a redox couple, the generation of which is controlled by metal ion release from metallothionein. PMID- 17134376 TI - Functional definition of the tobacco protoporphyrinogen IX oxidase substrate binding site. AB - PPO (protoporphyrinogen IX oxidase) catalyses the flavin-dependent six-electron oxidation of protogen (protoporphyrinogen IX) to form proto (protoporphyrin IX), a crucial step in haem and chlorophyll biosynthesis. The apparent K(m) value for wild-type tobacco PPO2 (mitochondrial PPO) was 1.17 muM, with a V(max) of 4.27 muM.min(-1).mg(-1) and a catalytic activity k(cat) of 6.0 s(-1). Amino acid residues that appear important for substrate binding in a crystal structure-based model of the substrate docked in the active site were interrogated by site directed mutagenesis. PPO2 variant F392H did not reveal detectable enzyme activity indicating an important role of Phe(392) in substrate ring A stacking. Mutations of Leu(356), Leu(372) and Arg(98) increased k(cat) values up to 100 fold, indicating that the native residues are not essential for establishing an orientation of the substrate conductive to catalysis. Increased K(m) values of these PPO2 variants from 2- to 100-fold suggest that these residues are involved in, but not essential to, substrate binding via rings B and C. Moreover, one prominent structural constellation of human PPO causing the disease variegate porphyria (N67W/S374D) was successfully transferred into the tobacco PPO2 background. Therefore tobacco PPO2 represents a useful model system for the understanding of the structure-function relationship underlying detrimental human enzyme defects. PMID- 17134377 TI - C-reactive protein modulates vagal heart rate control in patients with coronary artery disease. AB - Systemic inflammation is associated with sympathetic cardiac activation and decreased HRV (heart rate variability) in subjects at high risk of CAD (coronary artery disease). In the present study, we examined the influence of systemic inflammation, measured by CRP (C-reactive protein), on vagal HR (heart rate) control during behavioural relaxation in patients with CAD. It was hypothesized that CRP would be associated with decreased vagal HR modulation. Consecutive patients were screened 2 weeks prior to elective PTCA (percutaneous transluminal coronary angioplasty). The study was comprised of 29 subjects who represented the first and fourth quartiles of the CRP distribution: Low (0.47+/-0.07 microg/ml)- and High (8.19+/-1.95 microg/ml)-CRP groups respectively. Vagal HR control was quantified as RR high-frequency spectral power (0.15 to 0.40 Hz), and was assessed in log-transformed absolute units (logHF power). Near-IR particle immunoassay was used to determine high-sensitivity CRP concentration. Assessment entailed 5 min of silent reading and self-guided behavioural relaxation. RR logHF power was decreased in the High-CRP group across both assessment procedures (P=0.032). Behavioural relaxation increased RR logHF power for both the Low- and High-CRP groups (P=0.033). Hierarchical linear regression determined that CRP accounted for 18.9% of the variance in RR logHF power during behavioural relaxation (P=0.03), independent of baseline RR interval, cardiac medication, respiratory logHF power and body mass index. In conclusion, patients with CAD had augmented vagal HR control with behavioural relaxation, but this effect was moderated by the severity of CRP. Therefore it may be advisable to assess systemic inflammation in interventions aimed at improving neurocardiac regulation in patients with CAD. PMID- 17134378 TI - Editorial. PMID- 17134379 TI - Wound healing in the fetus. Possible role for inflammatory macrophages and transforming growth factor-beta isoforms. AB - Macrophages are believed to play a crucial role in wound healing by synthesizing and secreting numerous cytokines. Some of these cytokines, such as transforming growth factor-beta and tumor necrosis factor-alpha, promote fibrosis and repair. We have shown that macrophages are recruited to sterile fetal wounds and have the potential to regulate repair by synthesizing transforming growth factor-beta(1), transforming growth factor-beta(2), and tumor necrosis factor-alpha. Transforming growth factor-beta was present in fetal lamb wounds in higher amounts than in adult sheep wounds. Furthermore, the concentrations and ratios of the transforming growth factor-beta isoforms in wounds that healed without scarring were different from those in wounds that scarred; transforming growth factor beta(2) was highest in fetal wounds that did not scar and lowest in adult wounds. These data suggest that concentrations of transforming growth factor-beta isoforms rather than total transforming growth factor-beta concentration may be important in the regulation of fibrosis in prenatal and postnatal wound healing. PMID- 17134380 TI - Effects of basic fibroblast growth factor on the healing of partial-thickness donor sites. A prospective, randomized, double-blind trial. AB - A phase I/II clinical study was performed to evaluate the safety and potential efficacy of topical recombinant human basic fibroblast growth factor on the healing of partial-thickness skin graft donor sites in burned children. Each child served as his or her own control. In a blinded and random fashion, one donor site was sprayed with basic fibroblast growth factor (5 microg/cm(2)) on days 0 to 4 after harvest, whereas the other site was treated with vehicle. Twelve patients were entered in the study but one patient died of sepsis that was unrelated to growth factor treatment. Of the remaining 11 patients, no adverse events related to basic fibroblast growth factor occurred. Serum basic fibroblast growth factor levels were never detected and antibody levels remained unchanged. No differences in the rate of epithelialization or days until complete closure were noted (basic fibroblast growth factor = 12.9 +/- 3.9 days, placebo = 12.2 +/ 5.5 days; mean +/- standard error of the mean). No differences in pain, itching, wound fragility, erythema, scarring, or pigmentation were noted. All of the scars matured within 1 year with good to excellent results. Investigators, patients, or families could not distinguish between the two wounds. Although basic fibroblast growth factor proved safe, no enhancement of donor site healing was seen in this small study. Because the time for donor site healing limits subsequent autograft use in patients with sizeable burns, studies should focus on accelerating healing in patients with larger burns where donor site healing is delayed and reharvest is required. PMID- 17134381 TI - Development of a new wound dressing with antimicrobial delivery capability. AB - A bilaminar wound dressing composed of an outer membrane and an inner three dimensional matrix of a fabric or a sponge may be considered to constitute an ideal structure that promotes wound healing: the outer membrane prevents body fluid loss, controls water evaporation, and protects the wound surface from bacterial invasion, and the inner matrix encourages adherence by tissue growth into the matrix. Using this concept, we developed a biosynthetic wound dressing with a drug delivery capability. This medicated wound dressing is composed of a spongy sheet of a chitosane derivative and collagen mixture that is laminated to an antimicrobial-impregnated polyurethane membrane. In this study, a gentamycin sulfate-impregnated wound dressing was prepared and evaluated. The antimicrobial efficacy of this wound dressing was examined on an agar plate seeded with Pseudomonas aeruginosa. Also, the cytotoxicity of an antimicrobial released from this wound dressing was examined in an in vitro system with cultured skin substitutes. Both in vitro tests have shown that this wound dressing is capable of suppressing bacterial growth and minimizing cellular damage. In addition, in the treatment of wounds inflicted on rats and rabbits, this wound dressing was shown to be efficacious in covering full-thickness and split-thickness skin defects. Finally, the efficacy of this wound dressing was evaluated in a nonrandomized open-label study of 31 clinical cases. In 31 cases treated with this wound dressing, good or excellent wound healing was achieved. PMID- 17134382 TI - Effect of oxygen treatment and dressing oxygen permeability on wound healing. AB - Although it has been shown that both the level of tissue oxygen and its gradient are critical factors in the healing process, optimal conditions for oxygen therapy have not been determined. In this study, both the oxygen level and oxygen gradient for a full-thickness defect were modified on the basis of preceding in vitro studies to determine the effect on the healing process. The goal of this study was to help determine the optimal clinical oxygen treatment protocol. Specifically, the healing of full-thickness defects in a rabbit model as determined by histomorphometric analyses (cell and tissue volume fraction, epithelialization, and contraction) under two types of dressings with or without oxygen treatment (70% O(2)) was investigated. One of the dressings was more oxygen permeable than the other. No significant differences were found in the histomorphometric response between the wounds covered with the oxygen-permeable and oxygen-impermeable dressings in the group without supplemental oxygen. Oxygen treatment, however, seemed to enhance the healing response significantly. According to the histomorphometric response, the wounds covered with the oxygen impermeable dressings were significantly better than those with the oxygen permeable dressing in the oxygen-treated group after 1 week, but the wounds covered by the oxygen-permeable dressings were better healed at 3 weeks. Therefore, oxygen-impermeable dressings may be useful only in the early stages of healing, before granulation tissue formation. PMID- 17134383 TI - The hairless mouse ear: an in vivo model for studying wound neovascularization. AB - Microvascular ingrowth into damaged tissue is an essential component of the normal healing process. In fact, wound therapy is often aimed at promoting neovascularization. However, little is known about the mechanisms that regulate microvascular ingrowth into a healing wound. This limited knowledge is largely due to the lack of adequate models in which microvascular ingrowth can be quantitatively analyzed throughout the healing process. To address this deficiency, we developed a model in which a wound was created on the ear of the hairless mouse-a well established model for directly viewing and measuring skin microcirculation. While the animals were under ketamine and xylazine anesthesia, 2.25 mm diameter full-thickness wounds were created on the dorsum of hairless mouse ears down to but not including the cartilage (0.125 mm depth). With the use of video microscopy and computer-assisted digitized planimetry, the precise epithelial and neovascular wound edge was viewed and measured regularly throughout healing. Therefore, this model can provide objective data on wound epithelialization and neovascularization throughout healing. This model was used to examine the effect of topical wound agents on epithelialization and neovascularization. Differential effects by these anti-microbial agents on these two processes were observed, which suggests clinical implications for their use. PMID- 17134384 TI - Analysis of therapy-resistant venous leg ulcers. Can triple-layer treatment initiate healing? AB - Chronic leg ulcers represent a major health-care problem with considerable socioeconomic impact. Patients with seemingly therapy-resistant leg ulcers are common to all clinics. The purpose of the present study was to (1) examine a group of patients with nonhealing venous leg ulcers treated with a double-layer bandage and (2) evaluate whether the addition of an interactive hydrocolloid wound dressing could initiate healing in these patients. Twenty-two patients with ulcers caused by venous insufficiency were included. The patients had a mean ulcer duration of 27.6 years. Duration of the present ulcer was at least 1 year (mean >/= 4.1 years). Twenty of the 22 patients showed massive lipodermatosclerosis. Before inclusion, all patients had used double-layer bandage consisting of a zinc-impregnated bandage or stocking and a self-adhesive compression bandage for 1 year or longer without improvement. The new regimen was a triple-layer treatment with the hydrocolloid water applied over the ulcer and the traditional double-layer bandage unchanged. Three patients were dropped from the study. Nineteen patients were followed until healing or for 10 months. Nine of the 19 patients who completed the study healed. Ulcer area was reduced by 70% or greater in 7 patients and by 30% to 40% in two patients. One ulcer did not respond to the treatment and worsened slightly. The results of this study were encouraging and indicate that the triple-layer treatment with the hydrocolloid dressing applied to the ulcer should be evaluated in a randomized, controlled study in patients with less pessimistic prognoses. PMID- 17134385 TI - Effect of epidermal growth factor on cell proliferation in normal and wounded connective tissue. AB - Epidermal growth factor has been previously shown to stimulate connective tissue repair in the perforated rat mesentery. The mechanism by which epidermal growth factor accelerates closure of mesenteric perforations has not been established, but epidermal growth factor may stimulate mitosis, contraction, migration, or angiogenesis. In the present investigation, the effect of epidermal growth factor on connective tissue cell proliferation was studied during the initial phase of repair of mesenteric perforations and in unwounded mesentery. Laparotomies were performed on Sprague-Dawley rats, and standardized perforations were made with a scalpel in the center of the mesenteric "windows," leaving every second window as an internal control. Twice daily for 4 consecutive days, beginning on the day of surgery, the animals received by intraperitoneal injection either 10 microg of epidermal growth factor dissolved in phosphate-buffered saline solution or phosphate-buffered saline solution alone. Cell proliferation was measured by either mitotic index of fibroblasts and mesothelial cells or DNA content of individual fibroblast cell nuclei in the wound area or in unperforated control windows. Laparotomy alone was found to enhance proliferation during the early postoperative period, as shown by increased numbers of S + G2 fibroblasts and a greater mitotic index. Epidermal growth factor treatment increased the mitotic index in perforated windows on the third postoperative day, compared with controls treated with phosphate-buffered saline solution, but did not significantly increase either the number of S + G2 fibroblasts or the mitotic index in unwounded tissue. Also, the proliferative response after epidermal growth factor treatment was significantly higher in wounded tissue. This study shows that epidermal growth factor stimulates proliferation of connective tissue cells in wounded but not unwounded tissue, and such enhancement of fibroblast proliferation might be of importance in epidermal growth factor-stimulated connective tissue repair. PMID- 17134386 TI - Synergistic actions of platelet-derived growth factor and the insulin-like growth factors in vivo. AB - Topical application of growth factors has been shown to benefit both normal and impaired wound healing. In normal tissue repair, resident cells produce a "cocktail" of various types of growth factors that overlap in function. In vitro studies have proved that growth factor combinations can act synergistically to enhance cellular function beyond that achieved with individual growth factors. To determine whether similar combinations have a synergistic effect in vivo, we applied growth factor combinations topically to full-thickness skin wounds created in genetically diabetic mice. The C57BL/KsJ-db/db mouse is obese and has insulin-resistant diabetes, and it has been proved that this mouse has markedly impaired wound healing. Topical application of platelet-derived growth factor, insulin-like growth factor-I, or insulin-like growth factor-II enhances healing in this model. Marked synergism was found when platelet-derived growth factor and insulin-like growth factor-II were combined to produce augmentation in wound closure beyond that achieved by application of the individual growth factors. The synergistic effect allowed for improved tissue repair at doses of platelet derived growth factor and insulin-like growth factor-II that were ineffective when applied individually. The addition of insulin-like growth factor-I or insulin to platelet-derived growth factor produced no significant synergism. Because multiple growth factors are released in the wound during the healing process, it is not surprising that their combination further enhances healing. Growth factor combinations should become an important addition to the armamentarium for the treatment of chronic, nonhealing wounds. PMID- 17134387 TI - First Joint Meeting of The Wound Healing Society and the European Tissue Repair Society. RAI Congress Centre, Amsterdam, The Netherlands, August 22-25, 1993. PMID- 17134388 TI - Environmental and human health impacts of growing genetically modified herbicide tolerant sugar beet: a life-cycle assessment. AB - There is ongoing debate concerning the possible environmental and human health impacts of growing genetically modified (GM) crops. Here, we report the results of a life-cycle assessment (LCA) comparing the environmental and human health impacts of conventional sugar beet growing regimes in the UK and Germany with those that might be expected if GM herbicide-tolerant (to glyphosate) sugar beet is commercialized. The results presented for a number of environmental and human health impact categories suggest that growing the GM herbicide-tolerant crop would be less harmful to the environment and human health than growing the conventional crop, largely due to lower emissions from herbicide manufacture, transport and field operations. Emissions contributing to negative environmental impacts, such as global warming, ozone depletion, ecotoxicity of water and acidification and nutrification of soil and water, were much lower for the herbicide-tolerant crop than for the conventional crop. Emissions contributing to summer smog, toxic particulate matter and carcinogenicity, which have negative human health impacts, were also substantially lower for the herbicide-tolerant crop. The environmental and human health impacts of growing GM crops need to be assessed on a case-by-case basis using a holistic approach. LCA is a valuable technique for helping to undertake such assessments. PMID- 17134389 TI - Recombinant anti-hCG antibodies retained in the endoplasmic reticulum of transformed plants lack core-xylose and core-alpha(1,3)-fucose residues. AB - Plant-based expression systems are attractive for the large-scale production of pharmaceutical proteins. However, glycoproteins require particular attention as inherent differences in the N-glycosylation pathways of plants and mammals result in the production of glycoproteins bearing core-xylose and core-alpha(1,3)-fucose glyco-epitopes. For treatments requiring large quantities of repeatedly administered glycoproteins, the immunological properties of these non-mammalian glycans are a concern. Recombinant glycoproteins could be retained within the endoplasmic reticulum (ER) to prevent such glycan modifications occurring in the late Golgi compartment. Therefore, we analysed cPIPP, a mouse/human chimeric IgG1 antibody binding to the beta-subunit of human chorionic gonadotropin (hCG), fused to a C-terminal KDEL sequence, to investigate the efficiency of ER retrieval and the consequences in terms of N-glycosylation. The KDEL-tagged cPIPP antibody was expressed in transgenic tobacco plants or Agrobacterium-infiltrated tobacco and winter cherry leaves. N-Glycan analysis showed that the resulting plantibodies contained only high-mannose (Man)-type Man-6 to Man-9 oligosaccharides. In contrast, the cPIPP antibody lacking the KDEL sequence was found to carry complex N-glycans containing core-xylose and core-alpha(1,3)-fucose, thereby demonstrating the secretion competence of the antibody. Furthermore, fusion of KDEL to the diabody derivative of PIPP, which contains an N-glycosylation site within the heavy chain variable domain, also resulted in a molecule lacking complex glycans. The complete absence of xylose and fucose residues clearly shows that the KDEL-mediated ER retrieval of cPIPP or its diabody derivative is efficient in preventing the formation of non-mammalian complex oligosaccharides. PMID- 17134390 TI - T-DNA locus structure in a large population of soybean plants transformed using the Agrobacterium-mediated cotyledonary-node method. AB - Designing transformation experiments for either functional genomics or crop improvement requires knowledge of the transgene locus structure, number, transmission and expression resulting from a specific transformation method. We recently reported an improvement to the soybean [Glycine max (L.) Merrill] cotyledonary-node transformation method that resulted in the efficient production of transgenic plants. To characterize the transgene loci resulting from this method, we analysed 270 independent T0 plants and 95 randomly selected T1 progenies for T-DNA locus complexity using Southern analysis. The lines were transformed with Agrobacterium tumefaciens strains LBA4404 or EHA105 carrying the binary plasmids pGPTV, pTOK233, pCAMBIA1303 or pCAMBIA1309, and regenerated in medium supplemented with or without silver nitrate (AgNO3). Analysis in the T0 generation showed that the number of hpt-hybridizing fragments per plant ranged from 1-15, with 31.5% of the lines having a single hpt-hybridizing fragment. Each primary soybean transformant had, on average, 2.0 unlinked transgene loci and that half of the segregating loci in the T1 progenies were single, simple T-DNA insertions. Of the loci containing multiple T-DNA fragments, a low frequency had tandem and inverted repeat T-DNA structures. Integration of binary plasmid backbone sequences occurred in 37% of primary transformants. A. tumefaciens strain, binary plasmid and thiol treatment had no significant effect on transgene locus structure, numbers or expression. Interestingly, exposure of soybean explants to AgNO3 throughout shoot induction and elongation increased T-DNA locus complexity in the primary transformants and decreased silencing of gusA expression in the T1 generation. PMID- 17134391 TI - Bacteriophage T7 RNA polymerase-directed, inducible and tissue-specific over expression of foreign genes in transgenic plants. AB - A widely applicable bacteriophage T7 RNA polymerase-directed, tissue-specific and inducible over-expression of foreign genes in transgenic plants was developed. This was achieved through the simultaneous transformation of a modified T7 RNA polymerase to specifically transcribe the foreign gene placed under the control of T7 expression signals. The T7 RNA polymerase recognized the chimeric uidA gene integrated randomly into tobacco and rice genomes. Results from the use of six different promoters with different tissue specificities indicated that the recombinant protein was expressed at a several-fold (3-10-fold) higher level when compared with transgenes expressed directly under the control of these tissue specific promoters. An important feature of the T7 system in plants was the near uniform expression in the independently transformed plants, in contrast with the large variations observed in transgene expression under the direct control of plant promoters. In addition, our results demonstrated the application of the T7 system in the regulation of transgene expression through chemically inducible mechanisms. This versatility of controlled and regulated expression offers a powerful tool that could be used in various programmes in plant biotechnology and genomic studies. PMID- 17134392 TI - Field performance and starch characteristics of high-amylose potatoes obtained by antisense gene targeting of two branching enzymes. AB - Potato is an important crop for starch production, but there are limitations regarding the genetic variation of starch quality. In maize, starches with various properties have been available for a long time by mutational breeding. Amylose starch from potatoes differs from cereal amyloses in several functionally important aspects, such as a higher degree of polymerization. Areas of application in which the degree of polymerization is of importance include film forming and the polymeric properties of bioplastics. High-amylose potato lines have been achieved by inhibiting the two known branching enzyme forms of potato. A single inserted gene construct for the inhibition of both forms resulted in structural changes of the starch to levels of branching that were below the commercially available amylose standards of potato. The high-amylose potato lines were tested in multiple year field trials of agronomic performance and were used for the pilot plant production of starch. The introduced trait was confirmed to be stable over multiple years. The consequences of the modification were found to be an increased tuber yield, reduced starch content, smaller granule size and an increase in reducing sugars. PMID- 17134393 TI - Production of tailor-made fructans in sugar beet by expression of onion fructosyltransferase genes. AB - The consumption of fructans as a low caloric food ingredient or dietary fibre is rapidly increasing due to health benefits. Presently, the most important fructan source is chicory, but these fructans have a simple linear structure and are prone to degradation. Additional sources of high-quality tailor-made fructans would provide novel opportunities for their use as food ingredients. Sugar beet is a highly productive crop that does not normally synthesize fructans. We have introduced specific onion fructosyltransferases into sugar beet. This resulted in an efficient conversion of sucrose into complex, onion-type fructans, without the loss of storage carbohydrate content. PMID- 17134394 TI - Toxin-dependent utilization of engineered ribosomal protein L3 limits trichothecene resistance in transgenic plants. AB - The contamination of agricultural products with Fusarium mycotoxins is a problem of world-wide importance. Fusarium graminearum and related species, which are important pathogens of small grain cereals and maize, produce an economically important and structurally diverse class of toxins designated trichothecenes. Trichothecenes inhibit eukaryotic protein synthesis. Therefore, a proposed role for these fungal toxins in plant disease development is to block or delay the expression of defence-related proteins induced by the plant. Using yeast as a model system, we have identified several mutations in the gene encoding ribosomal protein L3 (Rpl3), which confer semi-dominant resistance to trichothecenes. Expression of an engineered tomato RPL3 (LeRPL3) cDNA, into which one of the amino acid changes identified in yeast was introduced, improved the ability of transgenic tobacco plants to adapt to the trichothecene deoxynivalenol (DON), but did not result in constitutive resistance. We show here that, in the presence of wild-type Rpl3 protein, the engineered Rpl3 protein is not utilized, unless yeast transformants or the transgenic plants are challenged with sublethal amounts of toxin. Our data from yeast two-hybrid experiments suggest that affinity for the ribosome assembly factor Rrb1p could be altered by the toxin resistance conferring mutation. This toxin-dependent utilization of the resistance conferring Rpl3 protein could seriously limit efforts to utilize the identified target alterations in transgenic crops to increase trichothecene tolerance and Fusarium resistance. PMID- 17134395 TI - Manipulating volatile emission in tobacco leaves by expressing Aspergillus nigerbeta-glucosidase in different subcellular compartments. AB - Expression of the Aspergillus nigerbeta-glucosidase gene, BGL1, in Nicotiana tabacum plants (cv. Xanthi) had a profound effect on the volatile emissions of intact and crushed leaves. BGL1 was expressed under the control of the cauliflower mosaic virus (CaMV) 35S promoter and targeted to the cytoplasm, cell wall, lytic vacuole (LV), chloroplast or endoplasmic reticulum (ER). Subcellular localization was confirmed by gold immunolabelling, followed by transmission electron microscopy (TEM). Significant beta-glucosidase activity was observed in transgenic plants expressing BGL1 in the cell wall, LV and ER. Compared with controls, all intact transgenic leaves were found to emit increased levels of 2 ethylhexanol, as determined by gas chromatography-mass spectrometry (GC-MS) analysis of the headspace volatiles. Plants expressing BGL1 in the cell wall (Tcw) emitted more trans-caryophyllene than did non-transgenic controls, whereas plants expressing BGL1 in the ER (Ter) and LV (Tvc) emitted more cembrene than did non-transgenic controls. Volatiles released from crushed transgenic leaves and collected with solid-phase microextraction (SPME) polydimethylsiloxane fibre were distinctly enhanced. Significant increases in linalool, nerol, furanoid cis linalool oxide, 4-methyl-1-pentanol, 6-methyl-hept-5-en-2-ol and 2-ethylhexanol were detected in transgenic plants when compared with wild-type controls. 3 Hydroxyl-beta-ionone levels were increased in crushed Tcw and Ter leaves, but were undetectable in Tvc leaves. The addition of glucoimidazole, a beta glucosidase inhibitor, abolished the increased emission of these volatiles. These results indicate that the expression of a fungal beta-glucosidase gene in different subcellular compartments has the potential to affect the emission of plant volatiles, and thereby to modify plant-environment communication and aroma of agricultural products. PMID- 17134396 TI - Genetic manipulation of gamma-linolenic acid (GLA) synthesis in a commercial variety of evening primrose (Oenothera sp.). AB - A robust Agrobacterium-mediated transformation procedure was developed for Rigel, a commercial cultivar of evening primrose, and used to deliver a cDNA encoding a Delta(6)-desaturase from borage under the control of a cauliflower mosaic virus (CaMV) 35S promoter. Analysis of the transformed plants demonstrated an altered profile of polyunsaturated fatty acids, with an increase in gamma-linolenic acid and octadecatetraenoic acid in leaf tissues when compared with control lines. PMID- 17134397 TI - Microarray analysis of nitric oxide responsive transcripts in Arabidopsis. AB - Nitric oxide (NO) is emerging as an important signalling molecule with diverse physiological functions in plants. In the current study, changes in gene expression in response to 0.1 mm and 1.0 mm sodium nitroprusside (SNP), a donor of NO, were studied in Arabidopsis using the whole genome ATH1 microarray, representing over 24,000 genes. We observed 342 up-regulated and 80 down regulated genes in response to NO treatments. These included 126 novel genes with unknown functions. Most of these changes were specific to NO treatment, as we observed a reverse trend when the plants were treated with NO scavenger, 2-[4 carboxyphenyl]-4,4,5,5-tetramethylimidazoline-1-oxy-3-oxide (c-PTIO). Hierarchical clustering revealed 162 genes showing a dose-dependent increase in signal from 0.1 mm SNP to 1.0 mm SNP treatment. We observed the up-regulation of several genes encoding disease-resistance proteins, WRKY proteins, transcription factors, zinc finger proteins, glutathione S-transferases, ABC transporters, kinases and biosynthetic genes of ethylene, jasmonic acid, lignin and alkaloids. This report provides an insight into the molecular basis for the seemingly diverse biological functions of NO in plants. Interestingly, about 2.0% of the genes in Arabidopsis responded to NO treatment, about 10% of which were transcription factors. NO may also influence the plant's signal transduction network as indicated by the transcriptional activation of several protein kinases, including a mitogen-activated protein (MAP) kinase. We identified many genes previously not shown to be associated with NO responses in plants, and this is the first report of NO responsive genes based on a whole genome microarray. PMID- 17134398 TI - Carbon metabolite sensing and signalling. AB - The regulation of carbon metabolism in plant cells responds sensitively to the levels of carbon metabolites that are available. The sensing and signalling systems that are involved in this process form a complex web that comprises metabolites, transporters, enzymes, transcription factors and hormones. Exactly which metabolites are sensed is not yet known, but candidates include sucrose, glucose and other hexoses, glucose-6-phosphate, trehalose-6-phosphate, trehalose and adenosine monophosphate. Important components of the signalling pathways include sucrose non-fermenting-1-related protein kinase-1 (SnRK1) and hexokinase; sugar transporters are also implicated. A battery of genes and enzymes involved in carbohydrate metabolism, secondary metabolism, nitrogen assimilation and photosynthesis are under the control of these pathways and fundamental developmental processes such as germination, sprouting, pollen development and senescence are affected by them. Here we review the current knowledge of carbon metabolite sensing and signalling in plants, drawing comparisons with homologous and analogous systems in animals and fungi. We also review the evidence for cross talk between carbon metabolite and other major signalling systems in plant cells and the prospects for manipulating this fundamentally important aspect of metabolic regulation for crop improvement. PMID- 17134399 TI - First-generation SNP/InDel markers tagging loci for pathogen resistance in the potato genome. AB - A panel of 17 tetraploid and 11 diploid potato genotypes was screened by comparative sequence analysis of polymerase chain reaction (PCR) products for single nucleotide polymorphisms (SNPs) and insertion-deletion polymorphisms (InDels), in regions of the potato genome where genes for qualitative and/or quantitative resistance to different pathogens have been localized. Most SNP and InDel markers were derived from bacterial artificial chromosome (BAC) insertions that contain sequences similar to the family of plant genes for pathogen resistance having nucleotide-binding-site and leucine-rich-repeat domains (NBS LRR-type genes). Forty-four such NBS-LRR-type genes containing BAC-insertions were mapped to 14 loci, which tag most known resistance quantitative trait loci (QTL) in potato. Resistance QTL not linked to known resistance-gene-like (RGL) sequences were tagged with other markers. In total, 78 genomic DNA fragments with an overall length of 31 kb were comparatively sequenced in the panel of 28 genotypes. 1498 SNPs and 127 InDels were identified, which corresponded, on average, to one SNP every 21 base pairs and one InDel every 243 base pairs. The nucleotide diversity of the tetraploid genotypes (pi = 0.72 x 10(-3)) was lower when compared with diploid genotypes (pi = 2.31 x 10(-3)). RGL sequences showed higher nucleotide diversity when compared with other sequences, suggesting evolution by divergent selection. Information on sequences, sequence similarities, SNPs and InDels is provided in a database that can be queried via the Internet. PMID- 17134400 TI - Foreign gene products can be enhanced by introduction into low storage protein mutants. AB - Transgenic rice expressing soybean glycinin in its endosperm was crossed with two types of low-glutelin mutants to determine how much storage the protein mutants can contribute to increases in glycinin accumulation. The glycinin level (102 microg/100 mg seed) in the parental transgenic line was enhanced to approximately 224-237 microg/100 mg seed within a genetic background deficient in glutelin (i.e. of low glutelins). The enrichment of this foreign gene product was compensated by a decrease in the expression of other endogenous prolamine and globulin storage proteins, resulting in an almost equivalent total amount of seed storage proteins. These results show that low storage protein mutants can provide potentially useful hosts for the expression of foreign genes, allowing a higher level accumulation, because they can provide wider space for the accumulation of foreign gene products than in the normal host plant. PMID- 17134401 TI - Development of an efficient cis-trans-cis ribozyme cassette to inactivate plant genes. AB - Inactivation of a targeted gene is one of the main strategies used to understand their precise cellular role. In plants, apart from chemical or physical mutagenesis and random insertions of DNA elements followed by screening for a desired phenotype, the most common strategy to inhibit the expression of a given gene involves RNA silencing. This can be achieved either through antisense suppression, sense over-expression leading to co-suppression, or expression of double-stranded DNA constructs (dsRNA). The use of ribozymes to inhibit gene product accumulation has only been occasionally attempted, mainly because of the more complex genetic engineering procedure involved, although the specificity of ribozymes can be an important factor when targeting close members of a gene family. We report here the development of a new cis-acting ribozyme cassette for the production of RNAs with desired termini. Attention to many details has been brought in order to provide a powerful procedure for plant application. For example, ultrastable GNRA tetraloops were substituted for both loops II and III of cis-acting hammerhead sequences, thereby favouring folding into the catalytically active structure that results in the self-cleavage of all transcripts. We demonstrate the usefulness of this cassette by producing a ribozyme that cleaves in trans, originally embedded in the cis-acting self cleaving cassette. The activity of the cis-trans-cis construct, was demonstrated both in vitro and in vivo, in transgenic plants with the specific cleavage of an mRNA encoding a 2-oxo-glutarate-dependant dioxygenase predominantly expressed in pistils tissues and in leaves, from the wild potato Solanum chacoense. PMID- 17134402 TI - Improved forage digestibility of tall fescue (Festuca arundinacea) by transgenic down-regulation of cinnamyl alcohol dehydrogenase. AB - Lignification of cell walls during plant development has been identified as the major factor limiting forage digestibility and concomitantly animal productivity. cDNA sequences encoding a key lignin biosynthetic enzyme, cinnamyl alcohol dehydrogenase (CAD), were cloned from the widely grown monocotyledonous forage species tall fescue (Festuca arundinacea Schreb.). Recombinant tall fescue CAD expressed in E. coli exhibited the highest V(max)/K(m) values when coniferaldehyde and sinapaldehyde were used as substrates. Transgenic tall fescue plants carrying either sense or antisense CAD gene constructs were obtained by microprojectile bombardment of single genotype-derived embryogenic suspension cells. Severely reduced levels of mRNA transcripts and significantly reduced CAD enzymatic activities were found in two transgenic plants carrying sense and antisense CAD transgenes, respectively. These CAD down-regulated transgenic lines had significantly decreased lignin content and altered ratios of syringyl (S) to guaiacyl (G), G to p-hydroxyphenyl (H) and S to H units. No significant changes in cellulose, hemicellulose, neutral sugar composition, p-coumaric acid and ferulic acid levels were observed in the transgenic plants. Increases of in vitro dry matter digestibility of 7.2-9.5% were achieved in the CAD down-regulated lines, thus providing a novel germplasm to be used for the development of grass cultivars with improved forage quality. PMID- 17134403 TI - Monoclonal C5-1 antibody produced in transgenic alfalfa plants exhibits a N glycosylation that is homogenous and suitable for glyco-engineering into human compatible structures. AB - Structural analysis of the N-glycosylation of alfalfa proteins was investigated in order to evaluate the capacity of this plant to perform this biologically important post-translational modification. We show that, in alfalfa, N-linked glycans are processed into a large variety of mature oligosaccharides having core xylose and core alpha(1,3)-fucose, as well as terminal Lewis(a) epitopes. In contrast, expression of the C5-1 monoclonal antibody in alfalfa plants results in the production of plant-derived IgG1 which is N-glycosylated by a predominant glycan having a alpha(1,3)-fucose and a beta(1,2)-xylose attached to a GlcNAc2Man3GlcNAc2 core. Since this core is common to plant and mammal N-linked glycans, it therefore appears that alfalfa plants have the ability to produce recombinant IgG1 having a N-glycosylation that is suitable for in vitro or in vivo glycan remodelling into a human-compatible plantibody. For instance, as proof of concept, in vitro galactosylation of the alfalfa-derived C5-1 mAb resulted in a homogenous plantibody harbouring terminal beta(1,4)-galactose residues as observed in the mammalian IgG. PMID- 17134404 TI - Transcriptional regulation of a pineapple polyphenol oxidase gene and its relationship to blackheart. AB - Two genes encoding polyphenol oxidase (PPO) were isolated from pineapple (Ananas comosus[L.] Merr. cv. Smooth Cayenne). Sequence analyses showed that both contained a single intron and encoded typical chloroplast-localized PPO proteins, the sequences of which corresponded to two pineapple PPO cDNAs, PINPPO1 and PINPPO2, recently described by Stewart et al. (2001). Southern blot analyses suggested that pineapple contained only two PPO genes. Analysis of expression of PINPPO1 promoter GUS fusion constructs showed this promoter had a low basal activity and was cold- and wound-inducible, consistent with known mRNA expression profiles. Striking homologies to gibberellin response complexes (GARC) were observed in sequences of both the PINPPO1 and PINPPO2 promoters. Transient assays in mature pineapple fruit and stable expression in transgenic tobacco showed that PINPPO1 promoter-GUS fusions were indeed gibberellin (GA) responsive. A role for the element within the putative GARCs in mediating GA-responsiveness of the PINPPO1 promoter was confirmed by mutational analysis. PINPPO2 was also shown to be GA-responsive by RT-PCR analysis. Mutant PINPPO1 promoter-GUS fusion constructs, which were no longer GA-inducible, showed a delayed response to cold induction in pineapple fruit in transient assays, suggesting a role for GA in blackheart development. This was supported by observations that exogenous GA(3) treatment induced blackheart in the absence of chilling. Sequences showing homology to GARCs are also present in some PPO promoters in tomato, suggesting that GA regulates PPO expression in diverse species. PMID- 17134405 TI - Changes in methylation during progressive transcriptional silencing in transgenic subterranean clover. AB - A transgenic line of subterranean clover (Trifolium subterraneum) containing a gene for a sulphur-rich sunflower seed albumin (ssa gene) and a gene conferring tolerance to the herbicide phosphinothricin (bar gene) was previously shown to stably express these genes as far as the T3 generation. In subsequent generations there was a progressive decline in the level of expression of both of these genes such that, by the T7 generation, the plants were almost completely susceptible to the herbicide and the mean level of sunflower seed albumin was reduced to 10-30% of the level in the T2 and T3 generations. The decline in SSA protein correlated closely with a decline in the level of ssa RNA. In vitro transcription experiments with nuclei isolated from plants of the early and late generations showed that the reduced mRNA level was associated with a reduced level of transcription of the ssa transgene. Transcription of the bar transgene was also reduced in the late generations. Bisulphite sequencing analysis showed that the decline in expression of the ssa gene between T3 and subsequent generations correlated closely with increased CpG methylation in the promoter, but not in the coding region. Analysis of the bar gene promoter showed that high levels of CpG methylation preceded the first detectable decline in expression of the bar gene by one generation, suggesting that methylation was not the direct cause of transgene silencing in these plants. PMID- 17134406 TI - The use of botulinum toxins in the management of pain and headache. PMID- 17134407 TI - Cancer pain: progress since the WHO guidelines. AB - In has been 15 years since the WHO Guidelines for the management of cancer pain were developed. This article reviews the guideline development and its effectiveness. Current trends in cancer care utilizing aggressive chemotherapeutic and surgical protocols lead to many patients living longer with advanced cancer and an attendant increase in pain. Future trends in cancer pain care are outlined. PMID- 17134408 TI - Pharmacology of peripheral analgesia. AB - Pain may begin in the periphery with activation of nociceptor transducers. The present article reviews the pharmacology of drug action at the level of the primary afferent by discussing the following: [1] agents which block transduction processes (vanilloids, sodium ion channel blockers, antiserotonergic agents, antipurinergic agents); [2] agents inhibiting the transducer site (opioids, cannabinoids, alpha adrenergic agents); [3] agents blocking transducer-based modulation processes (anti-inflammatories, antikinin agents, antitachykinins); and [4] agents which block primary afferent-related modification processes (antineurotrophins). There is a clear role for many of these agents in the treatment of inflammatory pain and they have potential benefits for neuropathic pain with peripheral triggers. PMID- 17134409 TI - Pain generators of the lumbar spine. AB - Different anatomical structures and pathophysiological functions can be responsible for lumbar pain, each producing a distinctive clinical profile. Pain can arise from the intervertebral disc, either acutely as a primary disc related disorder, or as result of the degradation associated with chronic internal disc disruption. In either case, greatest pain provocation will be associated with movements and functions in the sagittal plane. Lumbar pain can also arise from afflictions within the zygapophyseal joint mechanism, as result of synovitis or chondropathy. Either of these conditions will produce the greatest pain provocation during three-dimensional movements, due to maximal stress to either the synovium or joint cartilage. Finally, patients can experience different symptoms associated with irritation to the dural sleeve, dorsal root ganglion, or chemically irritated lumbar nerve root. Differential diagnosis of these conditions requires a thorough examination and provides information that can assist the clinician in selecting appropriate management strategies. PMID- 17134410 TI - The fascia iliaca block for postoperative pain relief after knee surgery. AB - The fascia iliaca nerve block provides excellent postoperative pain relief after knee surgery. It is easy to perform, needle insertion is not directly next to nerves or vessels, and it is associated with minimal side effects. Instructive case reports as well as a description and discussion of the technique are presented. PMID- 17134411 TI - Discogenic low back pain. PMID- 17134413 TI - Is mineral oil comedogenic? PMID- 17134412 TI - Welcome! To the journal of cosmetic dermatology. PMID- 17134414 TI - Relevance of vitamins C and E in cutaneous photoprotection. AB - Ultraviolet (UV) radiation-induced oxidative stress may result in acute and chronic photodamage. Based on the endogenous antioxidant system, the administration of antioxidants for scavenging reactive oxygen species might be a promising strategy in the prevention of UV-induced skin reactions. The relevance of the most common antioxidants, vitamins E and C, is reviewed focusing on topical and systemic photoprotective effects in animals and humans. Topically applied vitamin C induced significant photoprotective effects at concentrations of at least 10% in animals and humans, whereas a photoprotective effect has not been demonstrated by oral administration even at high doses in humans. Topical vitamin E reduced erythema, sunburn cells, chronic UV-B-induced skin damage, and photocarcinogenesis in the majority of the published studies, whereas only high doses of oral vitamin E may affect the response to UV-B in humans. Combination of vitamins C and E, partly with other photoprotective compounds, did increase the photoprotective effects dramatically compared to monotherapies. This synergistic interplay of several antioxidants should be taken into consideration in future research on cutaneous photoprotection. PMID- 17134415 TI - Idebenone: a new antioxidant - Part I. Relative assessment of oxidative stress protection capacity compared to commonly known antioxidants. AB - Topical applications of skin care products containing antioxidants have become increasingly popular. Numerous studies have elucidated the biological effects of these substances. General antiaging effects, anti-inflammatory properties, photoprotective properties, and prevention of ultraviolet (UV) immunosuppression have been documented. However, a standardized method to characterize and compare the properties and oxidative stress protection capacity of antioxidants was lacking. A multistep in vitro process utilizing a variety of biochemical and cell biological methods combined with in vivo studies was designed to compare the oxidative stress protective capacity of commonly used antioxidants. Data were presented for L-ascorbic acid, dl-alpha-tocopherol, kinetin, dl-alpha lipoic acid, ubiquinone, and idebenone. Methods included using UV-induced radical trapping/scavenging capacity measured by photochemiluminescence, pro-oxidative systems (LDL-CuSO(4), microsome-NADPH/ADP/Fe(3+)) with measurement of primary and secondary oxidation products, UVB irradiation of human keratinocytes, and in vivo evaluation, using the human sunburn cell (SBC) assay. Correlation and trends between in vitro and in vivo results were established, and the standardized test protocol was used to quantify oxidative stress protection capacity of antioxidants. Summarizing and totaling the data equally weighted for each oxidative stress study, the overall oxidative protection capacity scores of 95, 80, 68, 55, 52, and 41 were obtained for idebenone, dl-alpha tocopherol, kinetin, ubiquinone, L-ascorbic acid, and dl-alpha lipoic acid, respectively. The higher the score, the more effective the overall oxidative stress protection capacity of the antioxidant became. This multistep protocol may serve as a standard in investigating and comparing new putative antioxidants for topical use as well as a valuable tool to assess the anti-inflammatory properties, photoprotective properties, and prevention of UV immunosuppression of topical antioxidants. PMID- 17134416 TI - Treatment of periorbital wrinkles by repeated medium-depth chemical peels in dark skinned individuals. AB - BACKGROUND: The periorbital region serves as a barometer of chronological and environmental age and, as such, persons often seek its cosmetic rejuvenation. OBJECTIVE: This study was conducted to determine the efficacy and safety of repeated, medium-depth chemical peels in the treatment of periorbital wrinkles in dark-skinned individuals. SUBJECTS AND METHODS: A total of 12 subjects (10 women and 2 men) with fine and/or medium-sized wrinkles were included in the study, the ages ranged from 30 to 55 years with a mean of 41.25. Focal, medium-depth peels of periorbital area including crow's foot wrinkles were performed using a combination of Jessner's solution followed by the application of 35% trichloroacetic acid (TCA). The study involved a four-session regimen. To be included in this study, the subject completed at least two peeling sessions, 1 month apart, and returned for follow-up according to the research schedule. RESULTS: Improvement occurred in 11 of 12 subjects. Marked improvement occurred in four subjects (33%), moderate improvement in three (25%), mild improvement in another three (25%), minimal improvement in one subject (8%), and no response in one subject (8%). There were seven patients with fine and five with medium periorbital wrinkles. For statistical analysis, the no and minimum response groups were categorized as nonresponders. The improvement that ranged between marked to mild occurred in 100% of subjects with fine wrinkles and 60% of subjects with medium wrinkles. Mild side effects appeared in four subjects (33%). CONCLUSION: It can be concluded that fine and/or medium periorbital wrinkling responds favorably to repeated, medium-depth chemical peels even in dark-skinned individuals with few mild side effects. PMID- 17134417 TI - Efficacy of a new antidandruff thermophobic foam: a randomized, controlled, investigator-blinded trial vs. ketoconazole 2% scalp fluid. AB - BACKGROUND: Ketoconazole (K), zinc pyrithione (ZP), and salicylic acid (SA) are compounds active in the treatment of dandruff. A thermophobic foam formulation containing K 1%, ZP 0.5%, and SA 2% is now available. STUDY AIM: To compare the efficacy of thermo phobic foam with K 2% scalp fluid in the treatment of moderate to severe dandruff. Patients and methods In a randomized, prospective, parallel group, investigator-blinded, 4-week treatment trial, a total of 54 patients (mean age 43 +/- 8 years) were enrolled. Foam (F group) (n = 37) or K 2% scalp fluid (SF group) (n = 17) was applied daily for 7 days and twice weekly for 3 weeks thereafter. Clinical assessment of the total dandruff severity score (TDSS) was performed at baseline, after 2 and 4 weeks. The TDSS was calculated using a four grade quantitative score (0 = no dandruff; 3 = severe dandruff) dividing the scalp area into four zones and adding the single score for each area. RESULTS: At baseline, TDSS was 7.1 +/- 2 and 5.1 +/- 1 in the F and SF groups, respectively. At the end of treatment period, beneficial effects were observed in both groups. The TDSS was reduced to 2.5 +/- 1 in the F group and to 3.7 +/- 1.2 in the SF group (mean difference in favor of F group: -1.2; 95% CI: -0.57 to -1.8) (P = 0.0003). A complete or nearly complete resolution of dandruff was observed in 24 out of 37 (64%: 95%CI: 48-77%) in the F group and in 7 out of 17 (41%: 95% CI: 21 64%) in the SF group (P = 0.06 between groups). CONCLUSIONS: This new antidandruff thermophobic foam has shown to be more effective than ketoconazole 2% scalp fluid in the treatment of severe dandruff. PMID- 17134418 TI - Effective treatment of Futcher's lines with Q-switched alexandrite laser. AB - This is a case report of a young female patient who presented with pigmentary demarcation lines type A on the anterolateral aspect of both arms, which were satisfactorily treated with a Q-switched alexandrite laser with no adverse effects. PMID- 17134419 TI - Evaluation of the ultrasound influence in the cutaneous penetration of d panthenol: test in vitro. AB - BACKGROUND: d-panthenol is a popular additive in cosmetic and pharmaceutical preparations. However, in order for this vitamin to provide skin benefits, it must penetrate the stratum corneum. OBJECTIVE: To verify the penetrability of d panthenol in the skin and evaluate the effect of ultrasound on the cutaneous penetration of this vitamin. METHODS: The diffusion cell technique with pig skin as the membrane, distilled water as the receptor solution, and a hydrophilic d panthenol gel were used for the research. The experimental groups were gel + d panthenol (10%) and gel + d-panthenol (10%) + ultrasound. The receptor solution was collected at predetermined times and the amount of d-panthenol was determined by using a spectrophotometer at 406 nm. RESULTS: Ultrasound resulted in a statistically significant increase (P < 0.05) in the penetration of d-panthenol at 2, 60, and 240 min. CONCLUSION: d-panthenol penetration through the pig's skin is enhanced through the use of ultrasound. PMID- 17134420 TI - Use of intense pulsed light in the treatment of scars. AB - BACKGROUND: Reducing erythema and infiltration of inflamed, hypertrophic, and colloidal scars have been a challenge for healthcare providers. Peer-reviewed scientific data for intense pulsed light systems are lacking. OBJECTIVE: A chronicle of three patients who have participated in the treatment of inflamed, hypertrophic, and colloidal scars, using intense pulsed light. METHODS: Intense pulsed light with a selection of wavelengths, pulse durations, and energy densities was used on patients with inflamed, hypertrophic, and colloidal scars. RESULTS: A definite improvement in scar tissue was observed and achieved in all the cases. CONCLUSION: Intense pulsed light source with the correct outputs is an effective tool for the treatment and improvement of inflamed, hypertrophic, and colloidal scars. PMID- 17134421 TI - The natural history of androgenetic alopecia. AB - Androgenetic alopecia (AGA) is the most common cause of hair loss, affecting up to 80% of men and 50% of women in their lifetime. Genetic predisposition to the disease is well known but the responsible genes have not been identified. Polymorphism in the androgen receptor gene has been recently detected in AGA.(1) Although the role of androgens, and particularly dihydrotestosterone (DHT), in causing the disease has been established for a long time, the natural history of AGA is still not completely understood. This paper reviews recent data about natural progression of the disease, as well as factors that may interfere with its course and long-term prognosis. PMID- 17134423 TI - Parting thoughts on acne, skin aging, and smoking. PMID- 17134422 TI - After 30 years ... the future of hydroxyacids. PMID- 17134424 TI - Brittle nails, fragile nails. PMID- 17134425 TI - Botulinum toxin for periocular lines: the single-injection technique. AB - BACKGROUND: Botulinum is a well established treatment for facial wrinkles and hyperfunctional lines, including those at the periocular area. OBJECTIVE: To illustrate a simple periocular injection technique. It differs from the usual one since it is a single injection, instead of several. METHODS: Thirty consecutive patients were treated with this technique on one side, and the standard multipuncture technique on the contralateral side. Each patient, being their own control, received 10-12 Botox units to each side. The orbicularis oculi muscle was injected using two different types of needles: a standard 30 g 13 mm needle for the multipunctured side and a 30 g 25 mm needle for the single puncture side. Follow up assessments were made on days 7, 15 and 120. RESULTS: Good results were achieved in all patients. No relevant cosmetic difference was noticeable between the two sides. The pharmacological effect lasted about 5 months, with no difference between the two techniques. Both techniques were safe and caused the same amount of bruising. The single injection was preferred by patients. The single-injection technique, on the other hand, requires more skill and experience by the operator. CONCLUSIONS: The single-injection technique for periocular lines is a useful alternative to the standard multipuncture technique. It reduces patients' discomfort. PMID- 17134426 TI - Recrystallization in different sunscreen formulations after cutaneous application. AB - BACKGROUND: Data on the structure of sunscreens prior to and after application to the skin and the possible impact of these factors on their efficacy are still scant. AIMS: The microscopic structure of several commercial sunscreen products containing benzophenone-3 (BP-3) was analysed under a light microscope, prior to and 2 h after application of the products to the skin, and compared with various control preparations with or without BP-3. METHODS: All tested formulations were mounted on microscope slides, viewed under a light microscope and photographed. Samples were taken directly from original packages or from freshly prepared prescription formulations. Two hours after application to the skin, the samples were taken and processed for microscopy. RESULTS: In some commercial sunscreens numerous crystals were formed upon cutaneous application, whereas others contained crystals both prior to and after epicutaneous application. A single commercial product (a lamellar ambiphyl emulsion, SPF28) retained its regular structure throughout the study. Control preparations with or without BP-3 and/or octyl methoxycinnamate contained crystals after cutaneous application. CONCLUSIONS: Ingredients, most probably BP-3, in many commercial sunscreens are prone to recrystallization while on the skin which might interfere with their UV light-absorbing function. PMID- 17134427 TI - Nail fragility syndrome and its treatment. AB - For reasons of clarity, definitions are offered for strength, hardness, flexibility, brittleness and toughness of the nails. Six clinical types of nail fragility are delineated: longitudinal furrows and splitting (onychorrhexis), single longitudinal splitting, multiple crenellated splitting, lamellar splitting (onychoschizia), transverse splitting and nail friability. Changes may be observed in the keratin structure of fragile nails. Nail brittleness is usually 'environmental' in origin, but sometimes may be part of a nail dystrophy. Household daily chores are particularly damaging. Among the acquired general causes, hypochromic anaemia and sideropaenia, arthritic deformities of the distal joints, peripheral vascular impairment and endocrinopathies are the best known. Useful therapeutic approaches are updated. They entail protection with plastic gloves worn over light cotton glove linings, the use of nail hardeners composed of two main types of products: a modified nail varnish that functions as a base coat or a hardener, such as dimethyl urea, which overcomes the objections related to formaldehyde; a systemic drug, biotine, is still useful. PMID- 17134428 TI - Brittle nails. AB - Nail brittleness is a common complaint characterized by weak inelastic nails that split, flake and crumble. It may be a consequence of factors that alter nail plate production and/or factors that damage the already keratinised nail plate. It is often idiopathic. It can also be caused by many dermatological and systemic diseases, nutritional deficiencies, drugs and traumas. Environmental and occupational factors that produce progressive dehydration of the nail plate have an important role in nail brittleness. Treatment of brittle nails is often difficult. Preventative measures, together with oral supplementation of vitamins (especially biotin), oligo-elements and amino acids, can be useful in improving nail strength. Cosmetic treatment affords camouflage and a degree of protection. PMID- 17134429 TI - Sensitive skin: a complex and multifactorial syndrome. AB - Many people complain of discomfort after application of commonly used skin care products, particularly to the face. This hyperreactivity of the skin is a non immunologically mediated skin inflammation. It seems to be the result of an intolerance of the skin to various stimuli that are normally well tolerated. It is difficult to assess the prevalence of 'sensitive skin' and sensitive skin related cosmetic intolerance because of the many possible exogenous and endogenous factors that trigger or aggravate this multifactorial syndrome. A thorough history is essential. Sometimes patch testing is needed both to standard allergens and also to all the patients' cosmetics and skin care products. All cosmetics should be stopped and then reintroduced one by one, at intervals of one to two weeks. The number, type and frequency of the application of skin care products used in the final programme should remain limited. Some patients benefit from psychological or even psychiatric help. Management is usually difficult and all therapeutic measures should be undertaken with patience and tenacity. PMID- 17134430 TI - Photodamage of the skin: protection and reversal with topical antioxidants. AB - Controversy exists as to whether topical antioxidants can be effective in protecting against and reversing photodamage to the skin. Topical vitamins C and E, as well as topical selenium, protect skin against sunburn, suntan and skin cancer and also reverse the mottled pigmentation and wrinkles of photoageing. However, only certain forms of these labile antioxidants are stable and active after percutaneous absorption. For effective topical application, vitamin C must be non-esterified, acidic and optimally at 20% concentration; vitamin E must be the non-esterified isomer d-alpha-tocopherol at 2-5% concentration. Selenium is only percutaneously absorbed and active when applied topically as l selenomethionine, optimally at 0.02-0.05%. There are two great advantages in applying an active formulation of topical antioxidants to the skin. First, the skin attains far higher levels of each antioxidant than can be achieved by only taking these vitamins orally. The level of vitamin C attained in the skin by topical application is 20-40 times that achievable with oral vitamin C. With topical application, the concentration of vitamin E in the skin increases by a factor of 10.6 and selenium by a factor of 1.7. Second, topical application arms the skin with a reservoir of antioxidants that cannot be washed or rubbed off, a protection which stays in the skin for several days after application. PMID- 17134431 TI - Magnetic resonance imaging and spectroscopy of skin and subcutis. AB - Amongst in vivo skin imaging methods, magnetic resonance (MR) imaging and spectroscopy is of high interest not only for its ability to distinguish structures at a submillimetre scale but also for its ability to describe the physiology of the different skin layers through the measurement of their intrinsic MR parameters. High spatial resolution MR imaging allows the differentiation of epidermis, dermis and hypodermis. Cutaneous appendages such as hair follicles are also clearly visible. By measuring proton relaxation times and proton density, it is possible to go a step further in the description of skin physiology in vivo through the analysis of water-macromolecular interactions within the dermis as an example. Finally, quantification of water and lipid components in healthy and diseased adipose tissue by localized spectroscopy is also described. Even if MR imaging is of little interest for dermatological diagnosis, some preliminary studies have shown interesting results for preoperative staging, postoperative follow-up, and assessment of the efficacy of new dermatological products. In healthy skin, MR imaging and spectroscopy is a very promising method for the study of chronological and photoageing effects. PMID- 17134432 TI - Intravascular lasers in the treatment of varicose veins. AB - Historically, surgical treatments, such as high ligation or complete removal of an incompetent greater saphenous vein, were used to treat varicose veins resulting from saphenofemoral junction reflux. The relative lack of efficacy of these invasive methods, along with potential morbidity and significant patient downtime, has inspired the search for other treatments. Endovenous radiofrequency closure of the greater saphenous vein is effective and safe but its high cost, in terms of non-reusable catheters, and its slow withdrawal rate impair its practicality. A new technique for endovenous occlusion using endoluminal laser technology offers a less invasive alternative to ligation and stripping as well as a faster and less expensive method to treat varicose saphenous trunks and junctions. Initial clinical experience in several hundred patients shows a high degree of success with minimal side effects, most of which can be prevented or minimized by minor modifications of the technique. This paper reviews the use of an intravascular laser to destroy varicose veins. Various wavelengths including 810, 940, 980, 1064 and 1320 nm have been used to produce intravascular destruction of varicose veins. The 1320-nm intravascular laser with a motorized pull-back system appears to be the most efficient and reproducible system to effectively close and/or destroy an incompetent greater saphenous vein. PMID- 17134433 TI - Nail beauty. AB - Nail beauty depends firstly upon nail health. It is a truism to say that a beautiful nail is always a healthy nail. In addition, there are subjective and evolving aspects of nail beauty which are related to fashion. The texture of the nail affects its appearance and function. Soft or brittle nails are fragile. This results in unattractive longitudinal and horizontal splitting. Fragility is encouraged by wet work and excessive nail manicuring, especially by the removal of nail cosmetics. Critical to the aesthetic appeal of the nail is its shape. Most pleasing are nails are those that conform to the 'magic' ratio, in which the nail's length is approximately equal to its breadth, especially for the thumbs. The role of nail decoration and nail art in nail beauty is a subjective and evolving question of fashion. PMID- 17134434 TI - Traumatic nail split. AB - In nail split, the nail plate is divided into two parts. Trauma involving the nail matrix, the generator of the nail plate, can result in nail plate splitting which may take some months to become apparent. Photographic documentation of this condition in the literature had hitherto been lacking. PMID- 17134435 TI - Benign summer light eruption and polymorphic light eruption: genetic and functional studies suggest that a revised nomenclature is required. AB - New research indicates that polymorphic light eruption (PLE) is an autoimmune disease against an ultraviolet radiation-induced cutaneous antigen. PLE may even confer some protection against skin cancer later in life. This new information demands a reassessment of the precise nature and nomenclature of PLE. Benign summer light eruption (BSLE) (lucite estivale benigne) is the name used in continental Europe, and particularly France, to describe a clinically short lived, itchy, papular eruption particularly affecting young women after several hours of sunbathing at the beginning of summer or on sunny vacations. Clinically more prolonged forms of solar eruption, starting early in spring and persisting for long periods, have been known in France as polymorphic light eruption (PLE) (lucite polymorphe) ('European PLE'). Investigative studies, however, now suggest that BSLE and some cases of 'European PLE' are part of the same spectrum. In the Anglo-Saxon literature, they are lumped together as PLE ('Anglo-Saxon PLE'). The other cases of 'European PLE', which do not fall within the compass of 'Anglo Saxon PLE', are, in the Anglo-Saxon literature, classified as either actinic prurigo (AP) (a genetically determined, prolonged, excoriated form of Anglo-Saxon PLE), or chronic actinic dermatitis (CAD) (a sunlight-induced eczema precisely resembling allergic contact dermatitis, apparently to an ultraviolet radiation induced antigen). It is therefore proposed that: i. the European term BSLE be dropped and that these patients be reclassified within the spectrum of (Anglo Saxon) PLE, ii. the European use of the term PLE ('European PLE') be discontinued, iii. those previously diagnosed as having 'European PLE' be reclassified as (Anglo-Saxon) PLE, AP or CAD, as appropriate. The benefits of such a change in nomenclature would be twofold, firstly a uniformity of terminology and secondly, and more importantly, terminology would then correlate better with our recently improved understanding of the pathogenesis of these disorders. PMID- 17134436 TI - Insufficient sunlight may kill 45,000 Americans each year from internal cancer. PMID- 17134437 TI - Acne excoriee - a patient's view. PMID- 17134438 TI - Pyrrhodagger. PMID- 17134439 TI - Euripidesdagger. PMID- 17134441 TI - Lasers alleviate acne. AB - Propionibacterium acnes (P. acnes) and sebaceous glands are involved in the pathogenesis of acne. Although often effective, traditional therapies can have drawbacks, such as photosensitivity, other toxicities, polypharmacy and frequent dosing. Lasers have been studied to seek a solution that may overcome these disadvantages. Lasers and other light therapies target the wavelengths of the porphyrins in P. acnes to induce thermal damage that causes the bacterium's destruction. Pulsed-dye laser (PDL) or a system of light pulses and heat with wavelengths between 430 and 1100 nm are both efficacious. The 1450-nm diode laser targets sebaceous glands and is also effective. Topical indocyanine green (ICG) photodynamic therapy (PDT), using the near-infrared (NIR) laser, targets either P. acnes or sebaceous glands. ICG-PDT has fewer adverse effects than aminolevulinic acid-PDT. Lasers are still a relatively new therapy for acne and may be best used in an adjuvant role. PMID- 17134443 TI - The way of all flesh--the 21st Century. PMID- 17134444 TI - The origins of ESCAD and J Cos Derm. PMID- 17134445 TI - Long-pulsed ruby laser-assisted hair removal in male-to-female transsexuals. AB - BACKGROUND: Unwanted male-pattern pilosity is a heavy psychological burden and can cause distress for male-to-female transsexuals. Orchidectomy and oestrogen supplementation combined with antiandrogens fail to make hair disappear. AIMS: To study the effect of long-pulsed ruby laser treatment. RESULTS: Hair density was successfully abated on the beard and chest of three male-to-female transsexuals. A lingering effect over 6 months with a 50-90% hair density reduction was obtained. CONCLUSION: Photothermolysis using the long-pulsed ruby laser is a promising, well-tolerated method of hair removal in male-to-female transsexuals under oestrogen supplementation and antiandrogens. PMID- 17134446 TI - Phlebectomy--cosmetic indications. AB - Ambulatory phlebectomy is a safe, aesthetic, effective and economical operative technique, which enables venous extraction with a special hook, assuring complete and definitive eradication of the veins. Minimal skin incisions (1-3 mm), or even needle punctures, usually result in little or no scarring. This aesthetic method is particularly indicated to treat unsightly varicose veins, such as reticular veins (popliteal fold, lateral thigh and leg), groin pudendal veins, veins of the ankles and the dorsal venous network of the foot. Curettage of telangiectasias is a less well-known technique, but it is also effective in removing networks of thick blue spider veins. Ambulatory phlebectomy of body areas other than the lower limbs include dilated periorbital, temporal or frontal venous networks and venous dilatation of the abdomen, upper limbs or dorsa of the hands. PMID- 17134447 TI - Barrier creams--skin protectants: can you protect skin? AB - Barrier creams (BC) are used in hand care to protect the user against damage from surfactants and other irritants; their clinical value remains debatable; some reports indicate that inappropriate BC application might induce a deleterious rather than a beneficial effect. Since many cosmetic formulations contain 'skin protectants', we review concept, application, and efficacy from the pertinent scientific literature. PMID- 17134448 TI - Nail beauty therapy: an attractive enhancement or a potential hazard? AB - Nail coatings which harden upon evaporation and coatings that polymerize may produce some reactions at the site of application to the nail itself, and distant reactions when small amounts of nail cosmetics are transferred by the hand to other areas of the skin. Nail cosmetic hazards may be occupational, or accidental, especially in children. Individuals wearing artificial nails tend to wear their nails longer, and are more careful about their nails when washing their hands. The sanitary conditions for the application of artificial nails are therefore paramount in preventing nail infections. PMID- 17134449 TI - Bulimia and anorexia nervosa: cutaneous manifestations. AB - Bulimia and anorexia nervosa (BN, AN) are considered psychiatric disorders that have physical complications. Several factors may play a role in the onset of AN and BN, including a familial predisposition to these disorders, as well as individual personality characteristics. Dissatisfaction with body shape and an overwhelming desire to be thin are considered as risk factors for the development of eating disorders. Skin changes are characteristic and are cardinal diagnostic symptoms and pointers to the diagnosis of eating disorders. They are a consequence of starvation and/or malnutrition, self-induced vomiting, drug consumption and concomitant psychiatric illness. A careful examination of the skin can suggest a diagnosis of eating disorder. Cosmetic dermatologists, whose patients are mostly women, have a unique opportunity to detect these signs. This is very important as an early diagnosis will influence prognosis. PMID- 17134450 TI - The physical basis of cosmetic defects of the nail plate. AB - Cosmetic defects of the nail cover a range of changes. Some are variants of normal which are considered unattractive, others are part of the normal ageing process and some changes are manifestations of local or systemic disease. Interpretation of these changes relies on recognition of the specific characteristics in question and an understanding of the anatomy and biology of nails. In many instances, therapies are limited and explanation of the changes represents one of the most useful contributions the professional can make to the affected individual. This article covers some of the most common cosmetic defects of nail involving colour, surface, brittleness and behaviour of surrounding tissues. Understanding of the physical basis of these defects is limited in many cases, but current thinking is recorded. PMID- 17134451 TI - Are we still dying for a tan? AB - We suggest that the risks and benefits to individual and population health of different patterns of sunlight exposure are poorly characterised. Furthermore, we believe that the risks of excess exposure to sunlight in North European countries have been overstated and that the potential benefits of exposure to sunlight (particularly in moderation) have been largely ignored. We contend that a re appraisal of these risks and benefits is required to allow us to better formulate public health policy and to provide the public and individuals with appropriate advice. PMID- 17134456 TI - A bias against baldness. PMID- 17134466 TI - An updated interdisciplinary clinical pathway for CRPS: report of an expert panel. AB - The goal of treatment in patients with complex regional pain syndrome (CRPS) is to improve function, relieve pain, and achieve remission. Current guidelines recommend interdisciplinary management, emphasizing 3 core treatment elements: pain management, rehabilitation, and psychological therapy. Although the best therapeutic regimen or the ideal progression through these modalities has not yet been established, increasing evidence suggests that some cases are refractory to conservative measures and require flexible application of the various treatments as well as earlier consideration of interventions such as spinal cord stimulation (SCS). While existing treatment guidelines have attempted to address the comprehensive management of CRPS, all fail to provide guidance for contingent management in response to a sudden change in the patient's medical status. This paper reviews the current pathophysiology as it is known, reviews the purported treatments, and provides a modified clinical pathway (guideline) that attempts to expand the scope of previous guidelines. PMID- 17134467 TI - Disorders of the sacroiliac joint. AB - Controversies have surrounded the sacroiliac joint. The sacroiliac joint (SIJ) is a considerably complex and strong joint with limited mobility, mechanically serving as a force transducer and a shock absorber. Anatomical changes are seen in the SIJ throughout an individual's lifetime. The ligamentous system associated with the SIJ serves to enhance stability and offer proprioceptive feedback in context with the rich plexus of articular receptors. Stability in the SIJ is related to form and force closure. Movement in the SIJ is 3-D about an axis outside of the joint. The functional examination of the SIJ is related to a clinical triad. PMID- 17134468 TI - Fibromyalgia syndrome, idiopathic widespread persistent pain or syndrome of myalgic encephalomyelopathy (SME): what is its nature? AB - CONTEXT: Fibromyalgia is a disorder that is appearing more and more in the clinical practice but is poorly understood. OBJECTIVE: This paper attempts to look at all available and reliable data on these conditions and will outline current, scientifically sound understanding of these disorders, treatment modalities and future directions for research. It also attempts to analyze the social and cultural implications. DATA SOURCES/STUDY SELECTION: Various terms used to describe these syndromes are fibromyalgia (FMS), and myalgic encephalomyelitis (ME). PubMed search was performed. Pertinent articles published in past 25 years and The National Academy of Sciences colloquium on Neurobiology of Pain was also reviewed. In addition, news-articles in the lay press as well as the Internet were monitored for material posted by sufferers of these disorders. DATA EXTRACTION: Studies were reviewed for clinical presentations, history of these disorders, comorbid conditions, etiology, biochemical and microbiologic abnormalities, abnormalities found on neuroimaging and functional neurophysiologic techniques. In addition patient postings on the Internet and articles appearing in lay press were reviewed and social implications are discussed. DATA SYNTHESIS: Upon analysis of these materials the review was organized based on the quality of data and it's contextual scientific, cultural and political meaning for this disorder. This review appears to be pointing towards an entirely new paradigm in pain disorders; therefore, a hypothesis and future direction of research is repeatedly suggested. CONCLUSION: This study proposes an entirely new paradigm in these disorders based on scientific and cultural data. PMID- 17134469 TI - Radiofrequency neurolysis for facet arthropathy: a retrospective case series and review of the literature. AB - CONTEXT: Facet arthropathy is a common cause of spine-related pain. Typically resulting from spondylosis, trauma, including surgical trauma or post surgical stress is also a significant cause. Radiofrequency thermocoagulation or neurolysis may be an effective modality providing long-term improvement. OBJECTIVES: To evaluate the success rates for radiofrequency neurolysis for facet arthropathy in a large retrospective case series in a single pain practice setting. DESIGN: A retrospective case series involving chart reviews and patient follow-up visits or telephone contacts of radiofrequency neurolytic procedures performed for facet arthropathy over a 4-year period. SETTING: Private practice pain clinic with academic affiliation in Tulsa, OK. PARTICIPANTS: One hundred forty eight patients with confirmed facet arthropathy refractory to conservative measures underwent 230 radiofrequency neurolysis procedures and were followed for a minimum of 1 year post procedure. For cervical facet procedures: 63 patients (106 procedures); age range F: 27-84 years old; M: 33-65 years old. For lumbar facet procedures: 85 patients (124 facet procedures); age range F: 19-81 years old; M: 20-77 years old. MAIN OUTCOME MEASURES: After the radiofrequency procedure, patients were followed with periodic visits or telephone contacts. Outcome measures were McGill short form pain questionnaire, VAS pain scores, muscle spasm scores, tenderness, range of motion and patient subjective global responses. RESULTS: Subjective patient responses were graded as follows: excellent:greater than 70% improvement, good: 50% to 70% improvement, fair: 30% to 49%, and poor: less than 30%. One hundred six radiofrequency procedures were performed in the 63 cervical cases and 124 in the 85 lumbar cases with those patients who had good to excellent responses undergoing repeat procedures. Of the patients with cervical facet radiofrequency procedures, 38 (37%), 51 (48%), 4 (3%) and 13 (12%) had excellent, good, fair or poor responses, respectively. Of the lumbar facet radiofrequency cases, 37 (30%), 52 (41%), 13 (10%) and 22 (19%) had excellent, good, fair or poor responses, respectively. Excellent responders noted an average duration of 10.8 months (range 3-34 months before dropping below 70% improvement level) for cervical cases and 7.9 months (range 3-20) for lumbar. Good responders noted an average duration of 6.5 months (range 3-22 months before dropping below 50% improvement level) for cervical and 6.8 months (range 3-48) for lumbar radiofrequency procedures. No significant side effects were experienced (short-term neuritis was seen in 2 patients who had cervical and 1 who had lumbar RFTC, but resolved in each case after a few weeks). CONCLUSIONS: In summary, 85% of cervical and 71% of lumbar RFTC cases had at least a 50% improvement in symptoms for extended periods. RFTC of median branches for facet arthropathy is a safe and efficacious modality with the potential for long-term benefit. PMID- 17134470 TI - Bupivacaine induces transient neurological symptoms after subarachnoid block. AB - Bupivacaine is a long-acting amide local anesthetic that was introduced to clinical practice in the early 1980s. Since then it has been extensively used for both peripheral blocks as well as neuraxial blockade in concentrations varying from 0.125% to 0.75%. Despite the relatively narrow safety margin, bupivacaine has become the most frequently used local anesthetic in obstetric anesthesia. It is 95% metabolized in the liver and 5% excreted unchanged in urine. Bupivacaine cardiotoxicity is related to the total dose rather than to the concentration of bupivacaine administered. It produces a dose-dependent delay in the transmission of impulses through the cardiac conduction system by blocking sodium channels. Transient neurologic symptoms (TNS) defined as bilateral symmetrical pain in the lower back and buttocks with radiation to both lower extremities after 5% lidocaine spinal anesthetic was first described by Schneider in 1993. Several studies have failed to show TNS after spinal anesthesia using bupivacaine 0.5% or 0.75%. In the literature there is only 1 case report of TNS after spinal anesthesia using bupivacaine and morphine. The following report describes a case of TNS following spinal anesthesia with bupivacaine 0.75%. PMID- 17134471 TI - Palliative medicine--part 2. AB - With the recent release of the report from The Institute of Medicine regarding terminal patients, palliative medicine could receive much needed attention among medical specialties in the form of improving education, advanced research, and improved reimbursements. Thus, we continue our two-part series on palliative medicine. In this section, we discuss the assessment and management of symptoms like constipation, nausea/vomiting, hypercalcemia, and hydration. PMID- 17134472 TI - Triplane tissue Doppler imaging to evaluate mechanical dyssynchrony before and after cardiac resynchronization in a patient with congenitally corrected transposition of the great arteries. AB - We report the case of a 13-year-old girl with congenitally corrected transposition of the great arteries. Since the implantation of a conventional pacemaker for acquired complete atrioventricular block, the patient experienced increased heart failure symptoms. Using triplane tissue Doppler imaging, significant intraventricular dyssynchrony induced by unilateral pacing and associated with diminished exercise capacity was demonstrated. A biventricular pacemaker was successfully implanted transvenously, leading to synchronous activation of the systemic ventricle and improved exercise capacity. PMID- 17134473 TI - Mutational analysis of the zinc metalloprotease EmpA of Vibrio anguillarum. AB - The extracellular zinc metalloprotease, EmpA, is a putative virulence factor involved in pathogenicity of the fish pathogen Vibrio anguillarum. The 611-amino acid precursor of this enzyme is encoded by the empA gene. The residues His346, His350, Glu370, Glu347, His429, Tyr361 and Asp417 are highly conserved and putatively function together at the active site of the enzyme. In this study, empA was inserted into pET24d(+) and expressed in Escherichia coli strain BL21(DE3) as a 6 x His tagged protein (r-EmpA). All the conserved residues of EmpA mentioned above were individually mutated by site-directed mutagenesis and the mutants were also expressed (m-r-EmpAs). r-EmpA and m-r-EmpAs were purified, and assayed for their proteolytic activities with azocasein as the substrate and cytotoxicities on a flounder gill cell line. m-r-EmpAs that had been mutated at His346, His350, Glu370 and Glu347 almost completely lost their proteolytic activity and cytotoxicity, pointing towards the essential roles played by these residues. In contrast, those mutated at Tyr361, His429 and Asp417 still retained a partial proteolytic activity and cytotoxicity. Our results indicate that these conserved residues play important roles in enzymatic activity and that the proteolytic activity of the enzyme is involved in the pathogenesis of V. anguillarum PMID- 17134475 TI - Lexical adaptation of link grammar to the biomedical sublanguage: a comparative evaluation of three approaches. AB - BACKGROUND: We study the adaptation of Link Grammar Parser to the biomedical sublanguage with a focus on domain terms not found in a general parser lexicon. Using two biomedical corpora, we implement and evaluate three approaches to addressing unknown words: automatic lexicon expansion, the use of morphological clues, and disambiguation using a part-of-speech tagger. We evaluate each approach separately for its effect on parsing performance and consider combinations of these approaches. RESULTS: In addition to a 45% increase in parsing efficiency, we find that the best approach, incorporating information from a domain part-of-speech tagger, offers a statistically significant 10% relative decrease in error. CONCLUSION: When available, a high-quality domain part-of-speech tagger is the best solution to unknown word issues in the domain adaptation of a general parser. In the absence of such a resource, surface clues can provide remarkably good coverage and performance when tuned to the domain. The adapted parser is available under an open-source license. PMID- 17134476 TI - An environment for relation mining over richly annotated corpora: the case of GENIA. AB - BACKGROUND: The biomedical domain is witnessing a rapid growth of the amount of published scientific results, which makes it increasingly difficult to filter the core information. There is a real need for support tools that 'digest' the published results and extract the most important information. RESULTS: We describe and evaluate an environment supporting the extraction of domain-specific relations, such as protein-protein interactions, from a richly-annotated corpus. We use full, deep-linguistic parsing and manually created, versatile patterns, expressing a large set of syntactic alternations, plus semantic ontology information. CONCLUSION: The experiments show that our approach described is capable of delivering high-precision results, while maintaining sufficient levels of recall. The high level of abstraction of the rules used by the system, which are considerably more powerful and versatile than finite-state approaches, allows speedy interactive development and validation. PMID- 17134477 TI - Automatic recognition of topic-classified relations between prostate cancer and genes using MEDLINE abstracts. AB - BACKGROUND: Automatic recognition of relations between a specific disease term and its relevant genes or protein terms is an important practice of bioinformatics. Considering the utility of the results of this approach, we identified prostate cancer and gene terms with the ID tags of public biomedical databases. Moreover, considering that genetics experts will use our results, we classified them based on six topics that can be used to analyze the type of prostate cancers, genes, and their relations. METHODS: We developed a maximum entropy-based named entity recognizer and a relation recognizer and applied them to a corpus-based approach. We collected prostate cancer-related abstracts from MEDLINE, and constructed an annotated corpus of gene and prostate cancer relations based on six topics by biologists. We used it to train the maximum entropy-based named entity recognizer and relation recognizer. RESULTS: Topic classified relation recognition achieved 92.1% precision for the relation (an increase of 11.0% from that obtained in a baseline experiment). For all topics, the precision was between 67.6 and 88.1%. CONCLUSION: A series of experimental results revealed two important findings: a carefully designed relation recognition system using named entity recognition can improve the performance of relation recognition, and topic-classified relation recognition can be effectively addressed through a corpus-based approach using manual annotation and machine learning techniques. PMID- 17134478 TI - A critical review of PASBio's argument structures for biomedical verbs. AB - BACKGROUND: Propositional representations of biomedical knowledge are a critical component of most aspects of semantic mining in biomedicine. However, the proper set of propositions has yet to be determined. Recently, the PASBio project proposed a set of propositions and argument structures for biomedical verbs. This initial set of representations presents an opportunity for evaluating the suitability of predicate-argument structures as a scheme for representing verbal semantics in the biomedical domain. Here, we quantitatively evaluate several dimensions of the initial PASBio propositional structure repository. RESULTS: We propose a number of metrics and heuristics related to arity, role labelling, argument realization, and corpus coverage for evaluating large-scale predicate argument structure proposals. We evaluate the metrics and heuristics by applying them to PASBio 1.0. CONCLUSION: PASBio demonstrates the suitability of predicate argument structures for representing aspects of the semantics of biomedical verbs. Metrics related to theta-criterion violations and to the distribution of arguments are able to detect flaws in semantic representations, given a set of predicate-argument structures and a relatively small corpus annotated with them. PMID- 17134479 TI - Mapping data elements to terminological resources for integrating biomedical data sources. AB - BACKGROUND: Data integration is a crucial task in the biomedical domain and integrating data sources is one approach to integrating data. Data elements (DEs) in particular play an important role in data integration. We combine schema- and instance-based approaches to mapping DEs to terminological resources in order to facilitate data sources integration. METHODS: We extracted DEs from eleven disparate biomedical sources. We compared these DEs to concepts and/or terms in biomedical controlled vocabularies and to reference DEs. We also exploited DE values to disambiguate underspecified DEs and to identify additional mappings. RESULTS: 82.5% of the 474 DEs studied are mapped to entries of a terminological resource and 74.7% of the whole set can be associated with reference DEs. Only 6.6% of the DEs had values that could be semantically typed. CONCLUSION: Our study suggests that the integration of biomedical sources can be achieved automatically with limited precision and largely facilitated by mapping DEs to terminological resources. PMID- 17134480 TI - Recurrent insertion and duplication generate networks of transposable element sequences in the Drosophila melanogaster genome. AB - BACKGROUND: The recent availability of genome sequences has provided unparalleled insights into the broad-scale patterns of transposable element (TE) sequences in eukaryotic genomes. Nevertheless, the difficulties that TEs pose for genome assembly and annotation have prevented detailed, quantitative inferences about the contribution of TEs to genomes sequences. RESULTS: Using a high-resolution annotation of TEs in Release 4 genome sequence, we revise estimates of TE abundance in Drosophila melanogaster. We show that TEs are non-randomly distributed within regions of high and low TE abundance, and that pericentromeric regions with high TE abundance are mosaics of distinct regions of extreme and normal TE density. Comparative analysis revealed that this punctate pattern evolves jointly by transposition and duplication, but not by inversion of TE-rich regions from unsequenced heterochromatin. Analysis of genome-wide patterns of TE nesting revealed a 'nesting network' that includes virtually all of the known TE families in the genome. Numerous directed cycles exist among TE families in the nesting network, implying concurrent or overlapping periods of transpositional activity. CONCLUSION: Rapid restructuring of the genomic landscape by transposition and duplication has recently added hundreds of kilobases of TE sequence to pericentromeric regions in D. melanogaster. These events create ragged transitions between unique and repetitive sequences in the zone between euchromatic and beta-heterochromatic regions. Complex relationships of TE nesting in beta-heterochromatic regions raise the possibility of a co-suppression network that may act as a global surveillance system against the majority of TE families in D. melanogaster. PMID- 17134481 TI - Improving the use of research evidence in guideline development: 2. Priority setting. AB - BACKGROUND: The World Health Organization (WHO), like many other organisations around the world, has recognised the need to use more rigorous processes to ensure that health care recommendations are informed by the best available research evidence. This is the second of a series of 16 reviews that have been prepared as background for advice from the WHO Advisory Committee on Health Research to WHO on how to achieve this. OBJECTIVES: We reviewed the literature on priority setting for health care guidelines, recommendations and technology assessments. METHODS: We searched PubMed and three databases of methodological studies for existing systematic reviews and relevant methodological research. We did not conduct systematic reviews ourselves. Our conclusions are based on the available evidence, consideration of what WHO and other organisations are doing and logical arguments. KEY QUESTIONS AND ANSWERS: There is little empirical evidence to guide the choice of criteria and processes for establishing priorities, but there are broad similarities in the criteria that are used by various organisations and practical arguments for setting priorities explicitly rather than implicitly, WHAT CRITERIA SHOULD BE USED TO ESTABLISH PRIORITIES?: WHO has limited resources and capacity to develop recommendations. It should use these resources where it has the greatest chance of improving health, equity, and efficient use of healthcare resources. We suggest the following criteria for establishing priorities for developing recommendations based on WHO's aims and strategic advantages: Problems associated with a high burden of illness in low and middle-income countries, or new and emerging diseases. No existing recommendations of good quality. The feasibility of developing recommendations that will improve health outcomes, reduce inequities or reduce unnecessary costs if they are implemented. Implementation is feasible, will not exhaustively use available resources, and barriers to change are not likely to be so high that they cannot be overcome. Additional priorities for WHO include interventions that will likely require system changes and interventions where there might be a conflict in choices between individual and societal perspectives. WHAT PROCESSES SHOULD BE USED TO AGREE ON PRIORITIES?: The allocation of resources to the development of recommendations should be part of the routine budgeting process rather than a separate exercise. Criteria for establishing priorities should be applied using a systematic and transparent process. Because data to inform judgements are often lacking, unmeasured factors should also be considered- explicitly and transparently. The process should include consultation with potential end users and other stakeholders, including the public, using well constructed questions, and possibly using Delphi-like procedures. Groups that include stakeholders and people with relevant types of expertise should make decisions. Group processes should ensure full participation by all members of the group. The process used to select topics should be documented and open to inspection. SHOULD WHO HAVE A CENTRALISED OR DECENTRALISED PROCESS?: Both centralised and decentralised processes should be used. Decentralised processes can be considered as separate "tracks". Separate tracks should be used for considering issues for specific areas, populations, conditions or concerns. The rationales for designating special tracks should be defined clearly; i.e. why they warrant special consideration. Updating of guidelines could also be considered as a separate "track", taking account of issues such as the need for corrections and the availability of new evidence. PMID- 17134482 TI - Improving the use of research evidence in guideline development: 3. Group composition and consultation process. AB - BACKGROUND: The World Health Organization (WHO), like many other organisations around the world, has recognised the need to use more rigorous processes to ensure that health care recommendations are informed by the best available research evidence. This is the third of a series of 16 reviews that have been prepared as background for advice from the WHO Advisory Committee on Health Research to WHO on how to achieve this. OBJECTIVE: In this review we address the composition of guideline development groups and consultation processes during guideline development. METHODS: We searched PubMed and three databases of methodological studies for existing systematic reviews and relevant methodological research. We did not conduct systematic reviews ourselves. Our conclusions are based on the available evidence, consideration of what WHO and other organisations are doing and logical arguments. KEY QUESTIONS AND ANSWERS: What should be the composition of a WHO-panel that is set up to develop recommendations? The existing empirical evidence suggests that panel composition has an impact on the content of the recommendations that are made. There is limited research evidence to guide the exact composition of a panel. Based on logical arguments and the experience of other organisations we recommend the following: Groups that develop guidelines or recommendations should be broadly composed and include important stakeholders such as consumers, health professionals that work within the relevant area, and managers or policy makers. Groups should include or have access to individuals with the necessary technical skills, including information retrieval, systematic reviewing, health economics, group facilitation, project management, writing and editing. Groups should include or have access to content experts. To work well a group needs an effective leader, capable of guiding the group in terms of the task and process, and capable of facilitating collaboration and balanced contribution from all of the group members. Because many group members will not be familiar with the methods and processes that are used in developing recommendations, groups should be offered training and support to help ensure understanding and facilitate active participation.What groups should be consulted when a panel is being set up?We did not identify methodological research that addressed this question, but based on logical arguments and the experience of other organisations we recommend that as many relevant stakeholder groups as practical should be consulted to identify suitable candidates with an appropriate mix of perspectives, technical skills and expertise, as well as to obtain a balanced representation with respect to regions and gender. What methods should WHO use to ensure appropriate consultations? We did not find any references that addressed issues related to this question. Based on logical arguments and the experience of other organisations we believe that consultations may be desirable at several stages in the process of developing guidelines or recommendations, including: Identifying and setting priorities for guidelines and recommendations, commenting on the scope of the guidelines or recommendations, commenting on the evidence that is used to inform guidelines or recommendations, commenting on drafts of the guidelines or recommendations, commenting on plans for disseminating and supporting the adaptation and implementation of the guidelines or recommendations, key stakeholder organisations should be contacted directly whenever possible, consultation processes should be transparent and should encourage feedback from interested parties. PMID- 17134483 TI - What makes man human: thirty-ninth James Arthur lecture on the evolution of the human brain, 1970. AB - What makes man human is his brain. This brain is obviously different from those of nonhuman primates. It is larger, shows hemispheric dominance and specialization, and is cytoarchitecturally somewhat more generalized. But are these the essential characteristics that determine the humanness of man? This paper cannot give an answer to this question for the answer is not known. But the problem can be stated more specifically, alternatives spelled out on the basis of available research results, and directions given for further inquiry. My theme will be that the human brain is so constructed that man, and only man, feels the thrust to make meaningful all his experiences and encounters. Development of this theme demands an analysis of the brain mechanisms that make meaning-and an attempt to define biologically the process of meaning. In this pursuit of meaning a fascinating variety of topics comes into focus: the coding and recoding operations of the brain; how it engenders and processes information and redundancy; and, how it makes possible signs and symbols and prepositional utterances. Of these, current research results indicate that only in the making of propositions is man unique-so here perhaps are to be found the keynotes that compose the theme. PMID- 17134484 TI - What makes humanity humane. AB - Scientific and popular lore have promulgated a connection between emotion and the limbic forebrain. However, there are a variety of structures that are considered limbic, and disagreement as to what is meant by "emotion". This essay traces the initial studies upon which the connection between emotion and the limbic forebrain was based and how subsequent experimental evidence led to confusion both with regard to brain systems and to the behaviors examined. In the process of sorting out the bases of the confusion the following rough outlines are sketched: 1) Motivation and emotion need to be distinguished. 2) Motivation and emotion are processed by the basal ganglia; motivation by the striatum and related structures, emotion by limbic basal ganglia: the amygdala and related structures. 3) The striatum processes activation of readiness, both behavioral and perceptual; the amygdala processes arousal, an intensive dimension that varies from interest to panic. 4) Activation of readiness deals with "what to do?" Arousal deals with novelty, with "what is it?" 5) Thus both motivation and emotion are the proactive aspects of representations, of memory: motivation, an activation of readiness; emotion, a processing of novelty, a departure from the familiar. 6) The hippocampal-cingulate circuit deals with efficiently relating emotion and motivation by establishing dispositions, attitudes. 7) The prefrontal cortex fine-tunes motivation, emotion and attitude when choices among complex or ambiguous circumstances are made. PMID- 17134485 TI - Karl Pribram, The James Arthur lectures, and what makes us human. AB - BACKGROUND: The annual James Arthur lecture series on the Evolution of the Human Brain was inaugurated at the American Museum of Natural History in 1932, through a bequest from a successful manufacturer with a particular interest in mechanisms. Karl Pribram's thirty-ninth lecture of the series, delivered in 1970, was a seminal event that heralded much of the research agenda, since pursued by representatives of diverse disciplines, that touches on the evolution of human uniqueness. DISCUSSION: In his James Arthur lecture Pribram raised questions about the coding of information in the brain and about the complex association between language, symbol, and the unique human cognitive system. These questions are as pertinent today as in 1970. The emergence of modern human symbolic cognition is often viewed as a gradual, incremental process, governed by inexorable natural selection and propelled by the apparent advantages of increasing intelligence. However, there are numerous theoretical considerations that render such a scenario implausible, and an examination of the pattern of acquisition of behavioral and anatomical novelties in human evolution indicates that, throughout, major change was both sporadic and rare. What is more, modern bony anatomy and brain size were apparently both achieved well before we have any evidence for symbolic behavior patterns. This suggests that the biological substrate underlying the symbolic thought that is so distinctive of Homo sapiens today was exaptively achieved, long before its potential was actually put to use. In which case we need to look for the agent, perforce a cultural one, that stimulated the adoption of symbolic thought patterns. That stimulus may well have been the spontaneous invention of articulate language. PMID- 17134486 TI - Who's afraid of Homo sapiens? AB - Understanding how humans differ from other animals, as well as how we are like them, requires comparative investigations. For the purpose of documenting the distinctive features of humans, the most informative research involves comparing humans to our closest relatives-the chimpanzees and other great apes. Psychology and anthropology have maintained a tradition of empirical comparative research on human specializations of cognition. The neurosciences, by contrast, have been dominated by the model-animal research paradigm, which presupposes the commonality of "basic" features of brain organization across species and discourages serious treatment of species differences. As a result, the neurosciences have made little progress in understanding human brain specializations. Recent developments in neuroimaging, genomics, and other non invasive techniques make it possible to directly compare humans and nonhuman species at levels of organization that were previously inaccessible, offering the hope of gaining a better understanding of the species-specific features of the human brain. This hope will be dashed, however, if chimpanzees and other great ape species become unavailable for even non-invasive research. PMID- 17134487 TI - What makes us human: revisiting an age-old question in the genomic era. AB - In 1970, Karl Pribram took on the immense challenge of asking the question, what makes us human? Nearly four decades later, the most significant finding has been the undeniable realization of how incredibly subtle and fine-scaled the unique biological features of our species must be. The recent explosion in the availability of large-scale sequence data, however, and the consequent emergence of comparative genomics, are rapidly transforming the study of human evolution. The field of comparative genomics is allowing us to reach unparalleled resolution, reframing our questions in reference to DNA sequence--the very unit that evolution operates on. But like any reductionist approach, it comes at a price. Comparative genomics may provide the necessary resolution for identifying rare DNA sequence differences in a vast sea of conservation, but ultimately we will have to face the challenge of figuring out how DNA sequence divergence translates into phenotypic divergence. Our goal here is to provide a brief outline of the major findings made in the study of human brain evolution since the Pribram lecture, focusing specifically on the field of comparative genomics. We then discuss the broader implications of these findings and the future challenges that are in store. PMID- 17134488 TI - The changes in renal function after a single dose of intravenous furosemide in patients with compensated liver cirrhosis. AB - BACKGROUND: Patients with compensated Child-A cirrhosis have sub clinical hypovolemia and diuretic treatment could result in renal impairment. AIM: To evaluate the changes in renal functional mass as reflected by DMSA uptake after single injection of intravenous furosemide in patients with compensated liver cirrhosis. METHODS: Eighteen cirrhotic patients were divided in two groups; eight patients (group 1, age 56 +/- 9.6 yrs, Gender 5M/3F, 3 alcoholic and 5 non alcoholic) were given low intravenous 40 mg furosemide and ten other patients (group 2, age 54 +/- 9.9, Gender 6M/4F, 4 alcoholic and 6 non alcoholic) were given high 120 mg furosemide respectively. Renoscintigraphy with 100MBq Of Tc 99 DMSA was given intravenously before and 90 minutes after furosemide administration and SPECT imaging was determined 3 hours later. All patients were kept under low sodium diet (80mEq/d) and all diuretics were withdrawn for 3 days. 8-hours UNa exertion, Calculated and measured Creatinine clearance (CCT) were performed for all patients. RESULTS: Intravenous furosemide increased the mean renal DMSA uptake in 55% of patients with compensated cirrhosis and these changes persist up to three hours after injection. This increase was at the same extent in either low or high doses of furosemide. (From 12.8% +/- 3.8 to 15.2% +/- 2.2, p < 0.001 in Gr I as compared to 10.6% +/- 4.6 to 13.5% +/- 3.6 in Gr 2, p < 0.001). In 8 patients (45%, 3 pts from Gr 1 and 5 pts from Gr 2) DMSA uptake remain unchanged. The mean 8 hrs UNa excretion after intravenous furosemide was above 80 meq/l and was higher in Gr 2 as compared to Gr 1 respectively (136 +/- 37 meq/l) VS 100 +/- 36.6 meq/l, P = 0.05). Finally, basal global renal DMSA uptake was decreased in 80% of patients; 22.5 +/- 7.5% (NL > 40%), as compared to normal calculated creatinine clearance (CCT 101 +/- 26), and measured CCT of 87 +/- 30 cc/min (P < 0.001). CONCLUSION: A single furosemide injection increases renal functional mass as reflected by DMSA in 55% of patients with compensated cirrhosis and identify 45% of patients with reduced uptake and who could develop renal impairment under diuretics. Whether or not albumin infusion exerts beneficial effect in those patients with reduced DMSA uptake remains to be determined. PMID- 17134489 TI - Intrapartum epidural analgesia and breastfeeding: a prospective cohort study. AB - BACKGROUND: Anecdotal reports suggest that the addition of fentanyl (an opioid) to epidural analgesia for women during childbirth results in difficulty establishing breastfeeding. The aim of this paper is to determine any association between epidural analgesia and 1) breastfeeding in the first week postpartum and 2) breastfeeding cessation during the first 24 weeks postpartum. METHODS: A prospective cohort study of 1280 women aged > or = 16 years, who gave birth to a single live infant in the Australian Capital Territory in 1997 was conducted. Women completed questionnaires at weeks 1, 8, 16 and 24 postpartum. Breastfeeding information was collected in each of the four surveys and women were categorised as either fully breastfeeding, partially breastfeeding or not breastfeeding at all. Women who had stopped breastfeeding since the previous survey were asked when they stopped. RESULTS: In the first week postpartum, 93% of women were either fully or partially breastfeeding their baby and 60% were continuing to breastfeed at 24 weeks. Intrapartum analgesia and type of birth were associated with partial breastfeeding and breastfeeding difficulties in the first postpartum week (p < 0.0001). Analgesia, maternal age and education were associated with breastfeeding cessation in the first 24 weeks (p < 0.0001), with women who had epidurals being more likely to stop breastfeeding than women who used non pharmacological methods of pain relief (adjusted hazard ratio 2.02, 95% CI 1.53, 2.67). CONCLUSION: Women in this cohort who had epidurals were less likely to fully breastfeed their infant in the few days after birth and more likely to stop breastfeeding in the first 24 weeks. Although this relationship may not be causal, it is important that women at higher risk of breastfeeding cessation are provided with adequate breastfeeding assistance and support. PMID- 17134490 TI - Interleukin-4 induction of the CC chemokine TARC (CCL17) in murine macrophages is mediated by multiple STAT6 sites in the TARC gene promoter. AB - BACKGROUND: Macrophages (Mtheta) play a central role in the innate immune response and in the pathology of chronic inflammatory diseases. Macrophages treated with Th2-type cytokines such as Interleukin-4 (IL-4) and Interleukin-13 (IL-13) exhibit an altered phenotype and such alternatively activated macrophages are important in the pathology of diseases characterised by allergic inflammation including asthma and atopic dermatitis. The CC chemokine Thymus and Activation Regulated Chemokine (TARC/CCL17) and its murine homologue (mTARC/ABCD-2) bind to the chemokine receptor CCR4, and direct T-cell and macrophage recruitment into areas of allergic inflammation. Delineating the molecular mechanisms responsible for the IL-4 induction of TARC expression will be important for a better understanding of the role of Th2 cytokines in allergic disease. RESULTS: We demonstrate that mTARC mRNA and protein are potently induced by the Th2 cytokine, Interleukin-4 (IL-4), and inhibited by Interferon-gamma (IFN-gamma) in primary macrophages (Mtheta). IL-4 induction of mTARC occurs in the presence of PI3 kinase pathway and translation inhibitors, but not in the absence of STAT6 transcription factor, suggesting a direct-acting STAT6-mediated pathway of mTARC transcriptional activation. We have functionally characterised eleven putative STAT6 sites identified in the mTARC proximal promoter and determined that five of these contribute to the IL-4 induction of mTARC. By in vitro binding assays and transient transfection of isolated sites into the RAW 264.7 Mtheta cell-line, we demonstrate that these sites have widely different capacities for binding and activation by STAT6. Site-directed mutagenesis of these sites within the context of the mTARC proximal promoter revealed that the two most proximal sites, conserved between the human and mouse genes, are important mediators of the IL-4 response. CONCLUSION: The induction of mTARC by IL-4 results from cooperative interactions between STAT6 sites within the mTARC gene promoter. Significantly, we have shown that transfer of the nine most proximal mTARC STAT6 sites in their endogenous conformation confers potent (up to 130-fold) IL-4 inducibility on heterologous promoters. These promoter elements constitute important and sensitive IL-4-responsive transcriptional units that could be used to drive transgene expression in sites of Th2 inflammation in vivo. PMID- 17134491 TI - Impact on and use of health services by international migrants: questionnaire survey of inner city London A&E attenders. AB - BACKGROUND: Changing immigration trends pose new challenges for the UK's open access health service and there is considerable speculation that migrants from resource-poor countries place a disproportionate burden on services. Data are needed to inform provision of services to migrant groups and to ensure their access to appropriate health care. We compared sociodemographic characteristics and impact of migrant groups and UK-born patients presenting to a hospital A&E/Walk-In Centre and prior use of community-based General Practitioner (GP) services. METHODS: We administered an anonymous questionnaire survey of all presenting patients at an A&E/Walk-In Centre at an inner-city London hospital during a 1 month period. Questions related to nationality, immigration status, time in the UK, registration and use of GP services. We compared differences between groups using two-way tables by Chi-Square and Fisher's exact test. We used logistic regression modelling to quantify associations of explanatory variables and outcomes. RESULTS: 1611 of 3262 patients completed the survey (response rate 49.4%). 720 (44.7%) were overseas born, representing 87 nationalities, of whom 532 (73.9%) were new migrants to the UK (< or =10 years). Overseas born were over-represented in comparison to local estimates (44.7% vs 33.6%; p < 0.001; proportional difference 0.111 [95% CI 0.087-0.136]). Dominant immigration status' were: work permit (24.4%), EU citizens (21.5%), with only 21 (1.3%) political asylum seekers/refugees. 178 (11%) reported nationalities from refugee-generating countries (RGCs), eg, Somalia, who were less likely to speak English. Compared with RGCs, and after adjusting for age and sex, the Australians, New Zealanders, and South Africans (ANS group; OR 0.28 [95% CI 0.11 to 0.71]; p = 0.008) and the Other Migrant (OM) group comprising mainly Europeans (0.13 [0.06 to 0.30]; p = 0.000) were less likely to have GP registration and to have made prior contact with GPs, yet this did not affect mode of access to hospital services across groups nor delay access to care. CONCLUSION: Recently arrived migrants are a diverse and substantial group, of whom migrants from refugee-generating countries and asylum seekers comprise only a minority group. Service reorganisation to ensure improved access to community-based GPs and delivery of more appropriate care may lessen their impact on acute services. PMID- 17134492 TI - The pendulum test as a tool to evaluate passive knee stiffness and viscosity of patients with rheumatoid arthritis. AB - BACKGROUND: The pendulum test of Wartenberg is a technique commonly used to measure passive knee motion with the aim to assess spasticity. We used this test to evaluate changes of the knee angular displacement, passive stiffness and viscosity in rheumatoid arthritis patients. Stiffness and viscosity represent passive resistances to joint motion associated with the structural properties of the joint tissue and of the muscular-tendon complex. Stiffness can be considered an intrinsic property of the tissues to resist deformation, while viscosity is related to cohesive forces between adjacent layers of tissues. Both parameters may influence the joint range of motion affecting angular displacement. METHODS: Nine women with rheumatoid arthritis were compared with a group of healthy women. With the subject half-lying, the relaxed knee was dropped from near-full extension and the characteristics of the ensuring damped unsustained knee oscillation evaluated. The kinematics of leg oscillations was recorded using ultrasonic markers (Zebris CMS HS 10) and the kinetic data were calculated from kinematic and anthropometric measures. RESULTS: Knee stiffness significantly increased (p < 0.001) in patients with respect to the control group, while differences in viscosity were not significant. Moreover, the amplitudes of first knee flexion (the maximal flexion excursion after knee release) and first knee extension (the maximal extension excursion after the first knee flexion) were significantly decreased (p < 0.001). A regression analysis showed that disease severity correlated moderately with stiffness (R2 = 0.68) and first flexion (R2 = 0.78). Using a multivariate regression, we found that increasing stiffness was the main factor for the reduction of flexion and extension motions. CONCLUSION: We showed that the Wartenberg test can be considered a practical tool to measure mechanical changes of knee caused by rheumatoid arthritis. This novel application of Wartenberg test could be useful to follow up the effects of pharmacological and rehabilitative interventions in this disease. PMID- 17134493 TI - Simultaneity in the millisecond range as a requirement for effective shape recognition. AB - Neurons of the visual system are capable of firing with millisecond precision, and synchrony of firing may provide a mechanism for "binding" stimulus elements in the image for purposes of recognition. While the neurophysiology is suggestive, there has been relatively little behavioral work to support the proposition that synchrony contributes to object recognition. The present experiments examined this issue by briefly flashing dots that were positioned at the outer boundary of namable objects, similar to silhouettes. Display of a given dot lasted only 0.1 ms, and temporal proximity of dot pairs, and among dot pairs, was varied as subjects were asked to name each object. In Exp 1, where the display of dots pairs was essentially simultaneous (0.2 ms to show both), there was a linear decline in recognition of the shapes as the interval between pairs increased from 0 ms to 6 ms. Compared with performance at 0 ms of delay, even the 2 ms interval between pairs produced a significant decrease in recognition. In Exp 2 the interval between pairs was constant at 3 ms, and the interval between pair members was varied. Here also a linear decline was observed as the interval between pair members increased from 0 ms to 1.5 ms, with the difference between 0 ms and 0.5 ms being significant. Thus minimal transient discrete cues can be integrated for purposes of shape recognition to the extent that they are synchronously displayed, and coincidence in the millisecond and even submillisecond range is needed for effective encoding of image data. PMID- 17134494 TI - Best practice guidelines for the molecular genetic diagnosis of Type 1 (HFE related) hereditary haemochromatosis. AB - BACKGROUND: Hereditary haemochromatosis (HH) is a recessively-inherited disorder of iron over-absorption prevalent in Caucasian populations. Affected individuals for Type 1 HH are usually either homozygous for a cysteine to tyrosine amino acid substitution at position 282 (C282Y) of the HFE gene, or compound heterozygotes for C282Y and for a histidine to aspartic acid change at position 63 (H63D). Molecular genetic testing for these two mutations has become widespread in recent years. With diverse testing methods and reporting practices in use, there was a clear need for agreed guidelines for haemochromatosis genetic testing. The UK Clinical Molecular Genetics Society has elaborated a consensus process for the development of disease-specific best practice guidelines for genetic testing. METHODS: A survey of current practice in the molecular diagnosis of haemochromatosis was conducted. Based on the results of this survey, draft guidelines were prepared using the template developed by UK Clinical Molecular Genetics Society. A workshop was held to develop the draft into a consensus document. The consensus document was then posted on the Clinical Molecular Genetics Society website for broader consultation and amendment. RESULTS: Consensus or near-consensus was achieved on all points in the draft guidelines. The consensus and consultation processes worked well, and outstanding issues were documented in an appendix to the guidelines. CONCLUSION: An agreed set of best practice guidelines were developed for diagnostic, predictive and carrier testing for hereditary haemochromatosis and for reporting the results of such testing. PMID- 17134495 TI - Validation of the Edinburgh Postpartum Depression Scale in a population of puerperal women in Mexico. AB - BACKGROUND: The Edinburgh postnatal depression scale (EPDS) has been validated and used successfully in detecting postnatal depression in several language versions in a number of countries. However, there is not any Mexican version of the EPDS that had been validated. Therefore, we sought to validate a Spanish translated Mexican version of the EPDS in a population of puerperal Mexican women. METHODS: One hundred puerperal women within their three month postpartum period attending routine postnatal consultations in a public hospital in Durango City, Mexico participated in the study. The participants were divided into two groups: one group included 49 women with less than 4 weeks of postpartum, and the other group included 51 women within 4 to 13 weeks of postpartum. All participants submitted a Spanish translated Mexican version of the EPDS and were interviewed by a psychiatrist to assess major and minor depression by using DSM IV criteria. RESULTS: Out of the 49 women with less than 4 weeks of postpartum, 4 were found as suffering from major depression and none from minor depression by using the DSM-IV criteria. In this group of women we found that the best EPDS score for screening depression was 11/12. This threshold showed a sensitivity of 75% (95% CI: 63.8-86.2), a specificity of 93% (95% CI: 84.6-100), a positive predictive value of 50%, a negative predictive value of 97.6%, and an area under the curve of 0.84. While in the 51 women within 4 to 13 weeks of postpartum, 7 were found as suffering from major depression and 1 from minor depression by using the DSM-IV criteria. In this group we found that the best EPDS score for screening depression was 7/8. This threshold showed a sensitivity of 75% (95% CI: 66.1-83.9), a specificity of 84% (95% CI: 76.1-91.9), a positive predictive value of 46.2%, a negative predictive value of 94.7% and an area under the curve of 0.80. CONCLUSION: The Mexican version of the EPDS can be considered for screening depression in puerperal Mexican women whenever cut-off scores of 11/12 and 7/8 in women with less than 4 weeks and within 4 to 13 weeks of postpartum are used, respectively. PMID- 17134496 TI - Genome-wide loss of heterozygosity and copy number alteration in esophageal squamous cell carcinoma using the Affymetrix GeneChip Mapping 10 K array. AB - BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is a common malignancy worldwide. Comprehensive genomic characterization of ESCC will further our understanding of the carcinogenesis process in this disease. RESULTS: Genome-wide detection of chromosomal changes was performed using the Affymetrix GeneChip 10 K single nucleotide polymorphism (SNP) array, including loss of heterozygosity (LOH) and copy number alterations (CNA), for 26 pairs of matched germ-line and micro-dissected tumor DNA samples. LOH regions were identified by two methods- using Affymetrix's genotype call software and using Affymetrix's copy number alteration tool (CNAT) software--and both approaches yielded similar results. Non random LOH regions were found on 10 chromosomal arms (in decreasing order of frequency: 17p, 9p, 9q, 13q, 17q, 4q, 4p, 3p, 15q, and 5q), including 20 novel LOH regions (10 kb to 4.26 Mb). Fifteen CNA-loss regions (200 kb to 4.3 Mb) and 36 CNA-gain regions (200 kb to 9.3 Mb) were also identified. CONCLUSION: These studies demonstrate that the Affymetrix 10 K SNP chip is a valid platform to integrate analyses of LOH and CNA. The comprehensive knowledge gained from this analysis will enable improved strategies to prevent, diagnose, and treat ESCC. PMID- 17134497 TI - Pfarao: a web application for protein family analysis customized for cytoskeletal and motor proteins (CyMoBase). AB - BACKGROUND: Annotation of protein sequences of eukaryotic organisms is crucial for the understanding of their function in the cell. Manual annotation is still by far the most accurate way to correctly predict genes. The classification of protein sequences, their phylogenetic relation and the assignment of function involves information from various sources. This often leads to a collection of heterogeneous data, which is hard to track. Cytoskeletal and motor proteins consist of large and diverse superfamilies comprising up to several dozen members per organism. Up to date there is no integrated tool available to assist in the manual large-scale comparative genomic analysis of protein families. DESCRIPTION: Pfarao (Protein Family Application for Retrieval, Analysis and Organisation) is a database driven online working environment for the analysis of manually annotated protein sequences and their relationship. Currently, the system can store and interrelate a wide range of information about protein sequences, species, phylogenetic relations and sequencing projects as well as links to literature and domain predictions. Sequences can be imported from multiple sequence alignments that are generated during the annotation process. A web interface allows to conveniently browse the database and to compile tabular and graphical summaries of its content. CONCLUSION: We implemented a protein sequence-centric web application to store, organize, interrelate, and present heterogeneous data that is generated in manual genome annotation and comparative genomics. The application has been developed for the analysis of cytoskeletal and motor proteins (CyMoBase) but can easily be adapted for any protein. PMID- 17134498 TI - Genome-wide sequence and functional analysis of early replicating DNA in normal human fibroblasts. AB - BACKGROUND: The replication of mammalian genomic DNA during the S phase is a highly coordinated process that occurs in a programmed manner. Recent studies have begun to elucidate the pattern of replication timing on a genomic scale. Using a combination of experimental and computational techniques, we identified a genome-wide set of the earliest replicating sequences. This was accomplished by first creating a cosmid library containing DNA enriched in sequences that replicate early in the S phase of normal human fibroblasts. Clone ends were then sequenced and aligned to the human genome. RESULTS: By clustering adjacent or overlapping early replicating clones, we identified 1759 "islands" averaging 100 kb in length, allowing us to perform the most detailed analysis to date of DNA characteristics and genes contained within early replicating DNA. Islands are enriched in open chromatin, transcription related elements, and Alu repetitive elements, with an underrepresentation of LINE elements. In addition, we found a paucity of LTR retroposons, DNA transposon sequences, and an enrichment in all classes of tandem repeats, except for dinucleotides. CONCLUSION: An analysis of genes associated with islands revealed that nearly half of all genes in the WNT family, and a number of genes in the base excision repair pathway, including four of ten DNA glycosylases, were associated with island sequences. Also, we found an overrepresentation of members of apoptosis-associated genes in very early replicating sequences from both fibroblast and lymphoblastoid cells. These data suggest that there is a temporal pattern of replication for some functionally related genes. PMID- 17134499 TI - Characterization of ovine hepatic gene expression profiles in response to Escherichia coli lipopolysaccharide using a bovine cDNA microarray. AB - BACKGROUND: During systemic gram-negative bacterial infections, lipopolysaccharide (LPS) ligation to the hepatic Toll-like receptor-4 complex induces the production of hepatic acute phase proteins that are involved in the host response to infection and limit the associated inflammatory process. Identifying the genes that regulate this hepatic response to LPS in ruminants may provide insight into the pathogenesis of bacterial diseases and eventually facilitate breeding of more disease resistant animals. The objective of this research was to profile the expression of ovine hepatic genes in response to Escherichia coli LPS challenge (0, 200, 400 ng/kg) using a bovine cDNA microarray and quantitative real-time PCR (qRT-PCR). RESULTS: Twelve yearling ewes were challenged iv with E. coli LPS (0, 200, 400 ng/kg) and liver biopsies were collected 4-5 hours post-challenge to assess hepatic gene expression profiles by bovine cDNA microarray and qRT-PCR analyses. The expression of CD14, C3, IL12R, NRAMP1, SOD and IGFBP3 genes was down regulated, whereas the expression of ACTHR, IFNalphaR, CD1, MCP-1 and GH was increased during LPS challenge. With the exception of C3, qRT-PCR analysis of 7 of these genes confirmed the microarray results and demonstrated that GAPDH is not a suitable housekeeping gene in LPS challenged sheep. CONCLUSION: We have identified several potentially important genes by bovine cDNA microarray and qRT-PCR analyses that are differentially expressed during the ovine hepatic response to systemic LPS challenge. Their potential role in regulating the inflammatory response to LPS warrants further investigation. PMID- 17134500 TI - Indirect exclusion of four candidate genes for generalized progressive retinal atrophy in several breeds of dogs. AB - BACKGROUND: Generalized progressive retinal atrophy (gPRA) is a hereditary ocular disorder with progressive photoreceptor degeneration in dogs. Four retina specific genes, ATP binding cassette transporter retina (ABCA4), connexin 36 (CX36), c-mer tyrosin kinase receptor (MERTK) and photoreceptor cell retinol dehydrogenase (RDH12) were investigated in order to identify mutations leading to autosomal recessive (ar) gPRA in 29 breeds of dogs. RESULTS: Mutation screening was performed initially by PCR and single strand conformation polymorphism (SSCP) analysis, representing a simple method with comparatively high reliability for identification of sequence variations in many samples. Conspicuous banding patterns were analyzed via sequence analyses in order to detect the underlying nucleotide variations. No pathogenetically relevant mutations were detected in the genes ABCA4, CX36, MERTK and RDH12 in 71 affected dogs of 29 breeds. Yet 30 new sequence variations were identified, both, in the coding regions and intronic sequences. Many of the sequence variations were in heterozygous state in affected dogs. CONCLUSION: Based on the ar transmittance of gPRA in the breeds investigated, informative sequence variations provide evidence allowing indirect exclusion of pathogenetic mutations in the genes ABCA4 (for 9 breeds), CX36 (for 12 breeds), MERTK (for all 29 breeds) and RDH12 (for 9 breeds). PMID- 17134501 TI - Effect of infliximab on mRNA expression profiles in synovial tissue of rheumatoid arthritis patients. AB - We examined the gene expression profiles in arthroscopic biopsies retrieved from 10 rheumatoid arthritis patients before and after anti-TNF treatment with infliximab to investigate whether such profiles can be used to predict responses to the therapy, and to study effects of the therapy on the profiles. Responses to treatment were assessed using European League Against Rheumatism response criteria. Three patients were found to be good responders, five patients to be moderate responders and two patients to be nonresponders. The TNF-alpha status of the biopsies from each of the patients before treatment was also investigated immunohistochemically, and it was detected in biopsies from four of the patients, including all three of the good responders. The gene expression data demonstrate that all patients had unique gene expression signatures, with low intrapatient variability between biopsies. The data also revealed significant differences between the good responding and nonresponding patients (279 differentially expressed genes were detected, with a false discovery rate < 0.025). Among the identified genes we found that MMP-3 was significantly upregulated in good responders (log2 fold change, 2.95) compared with nonresponders, providing further support for the potential of MMP-3 as a marker for good responses to therapy. An even more extensive list of 685 significantly differentially expressed genes was found between patients in whom TNF-alpha was found and nonresponders, indicating that TNF-alpha could be an important biomarker for successful infliximab treatment. Significant differences were also observed between biopsies taken before and after anti-TNF treatment, including 115 differentially expressed genes in the good responding group. Interestingly, the effect was even stronger in the group in which TNF-alpha was immunohistochemically detected before therapy. Here, 1,058 genes were differentially expressed, including many that were novel in this context (for example, CXCL3 and CXCL14). Subsequent Gene Ontology analysis revealed that several 'themes' were significantly over-represented that are known to be affected by anti-TNF treatment in inflammatory tissue; for example, immune response (GO:0006955), cell communication (GO:0007154), signal transduction (GO:0007165) and chemotaxis (GO:0006935). No genes reached statistical significance in the moderately responding or nonresponding groups. In conclusion, this pilot study suggests that further investigation is warranted on the usefulness of gene expression profiling of synovial tissue to predict and monitor the outcome of rheumatoid arthritis therapies. PMID- 17134502 TI - Identification of the first intragenic deletion of the PITX2 gene causing an Axenfeld-Rieger Syndrome: case report. AB - BACKGROUND: Axenfeld-Rieger syndrome (ARS) is characterized by bilateral congenital abnormalities of the anterior segment of the eye associated with abnormalities of the teeth, midface, and umbilicus. Most cases of ARS are caused by mutations in the genes encoding PITX2 or FOXC1. Here we describe a family affected by a severe form of ARS. CASE PRESENTATION: Two members of this family (father and daughter) presented with typical ARS and developed severe glaucoma. The ocular phenotype was much more severe in the daughter than in the father. Magnetic resonance imaging (MRI) detected an aggressive form of meningioma in the father. There was no mutation in the PITX2 gene, determined by exon screening. We identified an intragenic deletion by quantitative genomic PCR analysis and characterized this deletion in detail. CONCLUSION: Our findings implicate the first intragenic deletion of the PITX2 gene in the pathogenesis of a severe form of ARS in an affected family. This study stresses the importance of a systematic search for intragenic deletions in families affected by ARS and in sporadic cases for which no mutations in the exons or introns of PITX2 have been found. The molecular genetics of some ARS pedigrees should be re-examined with enzymes that can amplify medium and large genomic fragments. PMID- 17134503 TI - Identification of a gonadotropin-releasing hormone receptor orthologue in Caenorhabditis elegans. AB - BACKGROUND: The Caenorhabditis elegans genome is known to code for at least 1149 G protein-coupled receptors (GPCRs), but the GPCR(s) critical to the regulation of reproduction in this nematode are not yet known. This study examined whether GPCRs orthologous to human gonadotropin-releasing hormone receptor (GnRHR) exist in C. elegans. RESULTS: Our sequence analyses indicated the presence of two proteins in C. elegans, one of 401 amino acids [GenBank: NP_491453; WormBase: F54D7.3] and another of 379 amino acids [GenBank: NP_506566; WormBase: C15H11.2] with 46.9% and 44.7% nucleotide similarity to human GnRHR1 and GnRHR2, respectively. Like human GnRHR1, structural analysis of the C. elegans GnRHR1 orthologue (Ce-GnRHR) predicted a rhodopsin family member with 7 transmembrane domains, G protein coupling sites and phosphorylation sites for protein kinase C. Of the functionally important amino acids in human GnRHR1, 56% were conserved in the C. elegans orthologue. Ce-GnRHR was actively transcribed in adult worms and immunoanalyses using antibodies generated against both human and C. elegans GnRHR indicated the presence of a 46-kDa protein, the calculated molecular mass of the immature Ce-GnRHR. Ce-GnRHR staining was specifically localized to the germline, intestine and pharynx. In the germline and intestine, Ce-GnRHR was localized specifically to nuclei as revealed by colocalization with a DNA nuclear stain. However in the pharynx, Ce-GnRHR was localized to the myofilament lattice of the pharyngeal musculature, suggesting a functional role for Ce-GnRHR signaling in the coupling of food intake with reproduction. Phylogenetic analyses support an early evolutionary origin of GnRH-like receptors, as evidenced by the hypothesized grouping of Ce-GnRHR, vertebrate GnRHRs, a molluscan GnRHR, and the adipokinetic hormone receptors (AKHRs) and corazonin receptors of arthropods. CONCLUSION: This is the first report of a GnRHR orthologue in C. elegans, which shares significant similarity with insect AKHRs. In vertebrates, GnRHRs are central components of the reproductive endocrine system, and the identification of a GnRHR orthologue in C. elegans suggests the potential use of C. elegans as a model system to study reproductive endocrinology. PMID- 17134504 TI - Hyperlactatemia during cardiopulmonary bypass: determinants and impact on postoperative outcome. AB - INTRODUCTION: Hyperlactatemia during cardiopulmonary bypass is relatively frequent and is associated with an increased postoperative morbidity. The aim of this study was to determine which perfusion-related factors may be responsible for hyperlactatemia, with specific respect to hemodilution and oxygen delivery, and to verify the clinical impact of hyperlactatemia during cardiopulmonary bypass in terms of postoperative morbidity and mortality rate. METHODS: Five hundred consecutive patients undergoing cardiac surgery with cardiopulmonary bypass were admitted to this prospective observational study. During cardiopulmonary bypass, serial arterial blood gas analyses with blood lactate and glucose determinations were obtained. Hyperlactatemia was defined as a peak arterial blood lactate concentration exceeding 3 mmol/l. Pre- and intraoperative factors were tested for independent association with the peak arterial lactate concentration and hyperlactatemia. The postoperative outcome of patients with or without hyperlactatemia was compared. RESULTS: Factors independently associated with hyperlactatemia were the preoperative serum creatinine value, the presence of active endocarditis, the cardiopulmonary bypass duration, the lowest oxygen delivery during cardiopulmonary bypass, and the peak blood glucose level. Once corrected for other explanatory variables, hyperlactatemia during cardiopulmonary bypass remained significantly associated with an increased morbidity, related mainly to a postoperative low cardiac output syndrome, but not to mortality. CONCLUSION: Hyperlactatemia during cardiopulmonary bypass appears to be related mainly to a condition of insufficient oxygen delivery (type A hyperlactatemia). During cardiopulmonary bypass, a careful coupling of pump flow and arterial oxygen content therefore seems mandatory to guarantee a sufficient oxygen supply to the peripheral tissues. PMID- 17134505 TI - Urokinase-type plasminogen activator supports liver repair independent of its cellular receptor. AB - BACKGROUND: The urokinase-type (uPA) and tissue-type (tPA) plasminogen activators regulate liver matrix remodelling through the conversion of plasminogen (Plg) to the active protease plasmin. Based on the efficient activation of plasminogen when uPA is bound to its receptor (uPAR) and on the role of uPA in plasmin mediated liver repair, we hypothesized that uPA requires uPAR for efficient liver repair. METHODS: To test this hypothesis, we administered one dose of carbon tetrachloride (CCl4) to mice with single or combined deficiencies of uPA, uPAR and tPA, and examined hepatic morphology, cellular proliferation, fibrin clearance, and hepatic proteolysis 2-14 days later. RESULTS: Absence of uPAR alone or the combined absence of uPAR and tPA had no impact on the resolution of centrilobular injury, but the loss of receptor-free uPA significantly impaired the clearance of necrotic hepatocytes up to 14 days after CCl4. In response to the injury, hepatocyte proliferation was normal in mice of all genotypes, except for uPAR-deficient (uPAR degrees) mice, which had a reproducible but mild decrease by 33% at day 2, with an appropriate restoration of liver mass by 7 days similar to experimental controls. Immunostaining and zymographic analysis demonstrated that uPA alone promoted fibrin clearance from centrilobular regions and efficiently activated plasminogen. CONCLUSION: uPA activates plasminogen and promotes liver matrix proteolysis during repair via a process that neither requires its receptor uPAR nor requires a contribution from its functional counterpart tPA. PMID- 17134506 TI - Perioperative strategy in colonic surgery; LAparoscopy and/or FAst track multimodal management versus standard care (LAFA trial). AB - BACKGROUND: Recent developments in large bowel surgery are the introduction of laparoscopic surgery and the implementation of multimodal fast track recovery programs. Both focus on a faster recovery and shorter hospital stay. The randomized controlled multicenter LAFA-trial (LAparoscopy and/or FAst track multimodal management versus standard care) was conceived to determine whether laparoscopic surgery, fast track perioperative care or a combination of both is to be preferred over open surgery with standard care in patients having segmental colectomy for malignant disease. METHODS/DESIGN: The LAFA-trial is a double blinded, multicenter trial with a 2 x 2 balanced factorial design. Patients eligible for segmental colectomy for malignant colorectal disease i.e. right and left colectomy and anterior resection will be randomized to either open or laparoscopic colectomy, and to either standard care or the fast track program. This factorial design produces four treatment groups; open colectomy with standard care (a), open colectomy with fast track program (b), laparoscopic colectomy with standard care (c), and laparoscopic surgery with fast track program (d). Primary outcome parameter is postoperative hospital length of stay including readmission within 30 days. Secondary outcome parameters are quality of life two and four weeks after surgery, overall hospital costs, morbidity, patient satisfaction and readmission rate. Based on a mean postoperative hospital stay of 9 +/- 2.5 days a group size of 400 patients (100 each arm) can reliably detect a minimum difference of 1 day between the four arms (alfa = 0.95, beta = 0.8). With 100 patients in each arm a difference of 10% in subscales of the Short Form 36 (SF-36) questionnaire and social functioning can be detected. DISCUSSION: The LAFA-trial is a randomized controlled multicenter trial that will provide evidence on the merits of fast track perioperative care and laparoscopic colorectal surgery in patients having segmental colectomy for malignant disease. PMID- 17134507 TI - Second site escape of a T20-dependent HIV-1 variant by a single amino acid change in the CD4 binding region of the envelope glycoprotein. AB - BACKGROUND: We previously described the selection of a T20-dependent human immunodeficiency virus type-1 (HIV-1) variant in a patient on T20 therapy. The fusion inhibitor T20 targets the viral envelope (Env) protein by blocking a conformational switch that is critical for viral entry into the host cell. T20 dependent viral entry is the result of 2 mutations in Env (GIA-SKY), creating a protein that undergoes a premature conformational switch, and the presence of T20 prevents this premature switch and rescues viral entry. In the present study, we performed 6 independent evolution experiments with the T20-dependent HIV-1 variant in the absence of T20, with the aim to identify second site compensatory changes, which may provide new mechanistic insights into Env function and the T20 dependence mechanism. RESULTS: Escape variants with improved replication capacity appeared within 42 days in 5 evolution cultures. Strikingly, 3 cultures revealed the same single amino acid change in the CD4 binding region of Env (glycine at position 431 substituted for arginine: G431R). This mutation was sufficient to abolish the T20-dependence phenotype and restore viral replication in the absence of T20. The GIA-SKY-G431R escape variant produces an Env protein that exhibits reduced syncytia formation and reduced cell-cell fusion activity. The escape variant was more sensitive to an antibody acting on an early gp41 intermediate, suggesting that the G431R mutation helps preserve a pre-fusion Env conformation, similar to T20 action. The escape variant was also less sensitive to soluble CD4, suggesting a reduced CD4 receptor affinity. CONCLUSION: The forced evolution experiments indicate that the premature conformational switch of the T20 dependent HIV-1 Env variant (GIA-SKY) can be corrected by a second site mutation in Env (GIA-SKY-G431R) that affects the interaction with the CD4 receptor. PMID- 17134508 TI - A case study evaluating the use of clozapine in depression with psychotic features. AB - The purpose of this case study was to use an evidence based medicine approach to work through an unusual way of treating a common problem. We looked at an example of an in-patient with severe refractory psychotic depression who had been resistant to treatment with a combination of antidepressant, antipsychotics, mood stabiliser, and concomitant ECT therapy. We then undertook a literature search for the use of clozapine in a patient with severe refractory depression. Although the resulting evidence was low level and thin, we felt on balance that a trial of clozapine was justified. We used a BPRS inventory to monitor her mood prior to commencing clozapine. Her mood and functional abilities were monitored as her clozapine was titrated upwards. Our patient showed a significant improvement in mood and functional abilities and a reduction in her BPRS score during this period. Her symptoms improved to the point where she was successfully discharged home on a combination of clozapine and an antidepressant. The improvement was sustained for a further two years. We thought this was an important case to highlight the limited evidence in using this successful form of treatment for a common clinical problem and that further research in this area was needed. PMID- 17134509 TI - Single-walled carbon nanotube (SWCNT)-induced interstitial fibrosis in the lungs of rats is associated with increased levels of PDGF mRNA and the formation of unique intercellular carbon structures that bridge alveolar macrophages in situ. AB - BACKGROUND: Nanotechnology is a rapidly advancing industry with many new products already available to the public. Therefore, it is essential to gain an understanding of the possible health risks associated with exposure to nanomaterials and to identify biomarkers of exposure. In this study, we investigated the fibrogenic potential of SWCNT synthesized by chemical vapor deposition using cobalt (Co) and molybdenum (Mo) as catalysts. Following a single oropharyngeal aspiration of SWCNT in rats, we evaluated lung histopathology, cell proliferation, and growth factor mRNAs at 1 and 21 days post-exposure. Comparisons were made to vehicle alone (saline containing a biocompatible nonionic surfactant), inert carbon black (CB) nanoparticles, or vanadium pentoxide (V2O5) as a known inducer of fibrosis. RESULTS: SWCNT or CB caused no overt inflammatory response at 1 or 21 days post-exposure as determined by histopathology and evaluation of cells (>95% macrophages) in bronchoalveolar lavage (BAL) fluid. However, SWCNT induced the formation of small, focal interstitial fibrotic lesions within the alveolar region of the lung at 21 days. A small fraction of alveolar macrophages harvested by BAL from the lungs of SWCNT exposed rats at 21 days were bridged by unique intercellular carbon structures that extended into the cytoplasm of each macrophage. These "carbon bridge" structures between macrophages were also observed in situ in the lungs of SWCNT exposed rats. No carbon bridges were observed in CB-exposed rats. SWCNT caused cell proliferation only at sites of fibrotic lesion formation as measured by bromodeoxyuridine uptake into alveolar cells. SWCNT increased platelet-derived growth factor (PDGF)-A, PDGF-B, and PDGF-C mRNA levels significantly at 1 day as measured by Taqman quantitative real-time RT-PCR. At 21 days, SWCNT did not increase any mRNAs evaluated, while V2O5 significantly increased mRNAs encoding PDGF-A, -B, and -C chains, PDGF-R alpha, osteopontin (OPN), connective tissue growth factor (CTGF), and transforming growth factor (TGF)-beta1. CONCLUSION: Our findings indicate that SWCNT do not cause lung inflammation and yet induce the formation of small, focal interstitial fibrotic lesions in the alveolar region of the lungs of rats. Of greatest interest was the discovery of unique intercellular carbon structures composed of SWCNT that bridged lung macrophages. These "carbon bridges" offer a novel and easily identifiable biomarker of exposure. PMID- 17134510 TI - Detection of cytokine protein expression in mouse lung homogenates using suspension bead array. AB - BACKGROUND: The objective for this present study was to determine whether or not suspension bead array is a feasible method to detect changes in cytokine protein expression in mouse lung tissue homogenates. Here, we report on suspension bead array as a feasible method for detection of lipopolysaccharide (LPS)-evoked changes in cytokine protein expression in mouse lung tissue homogenates. MATERIALS AND METHODS: Mice were treated (0.2 ml, intraperitoneal, i.p.) with phosphate buffered saline (PBS) or LPS (0.25 mg/ml) and sacrificed at either 2- or 24-hours post treatment. Formalin-fixed and paraffin-embedded lung sections were evaluated by light microscopy. Flash frozen lung tissues were homogenized for measurement of various cytokine protein expression levels using suspension bead array, antibody array and ELISA. Comparison between groups was performed using a Wilcoxon signed rank test. RESULTS: Pulmonary perivascular edema and an accumulation of mixed cell infiltrates within blood and lymphatic vessels, as well as in the adjacent interstitium, were present at both 2- and 24-hours following LPS treatment. A minimal increase in the number of alveolar macrophages was also observed in the 24-hour LPS-treated mice only. The suspension bead array assay revealed statistically significant increases in mouse lung tissue homogenate levels of interleukin-6 (IL-6) and granulocyte/macrophage colony stimulating factor (GM-CSF) proteins and a decrease in IL-2 protein at 24-hours post LPS-treatment only. Similar cytokine protein expression patterns were observed using antibody array. Significantly increased IL-6 protein expression levels were also detected using ELISA, which correlated with the suspension bead array data. CONCLUSION: The present study shows that suspension bead array is a feasible method to detect changes in cytokine protein expression in mouse lung tissue homogenates. PMID- 17134511 TI - Ageing is associated with a decline in peripheral blood CD56bright NK cells. AB - BACKGROUND: Natural killer (NK) cells are cytotoxic lymphocytes that lack CD3 and express variable levels of CD16, CD56 and CD57. In recent years NK cells have been categorised into two major groups based on the level of CD56 expression. This phenotypic classification correlates with functional activity as CD56bright NK cells are the major cytokine producing subset whereas CD56dim NK cells exhibit greater cytotoxic activity. Previous studies have revealed a reduction in total NK cell numbers in association with ageing and this study sought to determine the potential influence of ageing on the number of NK cell subsets within peripheral blood. RESULTS: The number of NK (CD56+CD3-) cells within peripheral blood did not change with increasing age. The number of CD56dim NK cells also remained stable with ageing. In contrast the absolute number of CD56bright NK cells within peripheral blood declined by 48% with ageing from a mean of 15.6/microl in individuals aged 20-40 years to 8.1/microl in those aged 60+ years (p = 0.0004). CONCLUSION: The number of CD56bright NK cells within peripheral blood declines with age. As this population plays a central role in cytokine secretion during the innate immune response this decline may contribute to impaired immune regulation in elderly individuals. PMID- 17134512 TI - Alcohol use and abuse in training conscripts of the Hellenic navy. AB - OBJECTIVES: Alcohol abuse and addiction are big current problems of the developed world having multivariate causality and multiple effects. Alcohol abuse in young people is a matter of central importance due to its wide range long lasting effects, especially so in Greece where the problem has only recently started growing. The Hellenic Navy is interested in the complications of alcohol abuse in training conscripts. Because young conscripts will be placed in demanding positions, but also because in Greece the military service is obligatory and represents an important period for the socialization of young men. METHODS: In the present study, levels of alcohol use and abuse were measured in a sample of 650 male training conscripts of the Hellenic Navy. The tools used are: (a) two questionnaires measuring frequency and quantity of alcohol consumption and psychosocial variables, (b) the CAGE test, which is a questionnaire measuring hidden alcoholism. RESULTS: 38,1% conscripts were characterized problematic drinkers according the adolescents criteria. Additional psychological complications were related to alcohol use. Using the stricter criterion for adults (plus psychological complications) 8.9% were found to be problematic drinkers. The use of CAGE questionnaire which is measuring hidden alcoholism, identified 16% of the total sample as hidden alcoholics. DISCUSSION: The findings regarding unregular levels of alcohol use and abuse are presented as well as their relation to psychosocial complications and to demographic characteristics. The results are discussed in the light of Creek and international bibliography. PMID- 17134513 TI - Does listening to the sound of yourself chewing increase your enjoyment of food? AB - BACKGROUND: Anecdotal evidence suggests that listening to oneself eating results in a more pleasurable eating experience. Maximising the sensory experience of eating can result in increased oral intake and is potentially valuable in improving nutritional status in at-risk patients. OBJECTIVE: This pilot study investigates the association between listening to the sound of oneself eating and the consequences on enjoyment of eating. DESIGN: Prospective, randomized, controlled, cross-over trial of 10 fit, adult volunteers. Participants were timed eating a standardised amount of bread, and were randomized to eat in silence or whilst listening to their own amplified chewing and swallowing. Measurements of pulse and blood pressure were recorded throughout the procedure. Subjective pleasure scores were documented and the procedure repeated in the alternate study arm. RESULTS: There was no significant relationship demonstrated between listening to oneself chewing and the enjoyment of eating. CONCLUSION: Although this small pilot study was unable to demonstrate a significant relationship between listening to oneself chewing and enjoyment of eating, other evidence suggests that distraction techniques have a beneficial effect on dietary intake. Such techniques can be applied in a clinical setting and further work in this area has valuable potential. PMID- 17134514 TI - Association study of the vesicular monoamine transporter 1 (VMAT1) gene with schizophrenia in a Japanese population. AB - BACKGROUND: Vesicular monoamine transporters (VMATs) mediate accumulation of monoamines such as serotonin, dopamine, adrenaline, and noradrenaline from the cytoplasm into storage organelles. The VMAT1 (alternatively solute carrier family 18: SLC18A1) regulates such biogenic amines in neuroendocrine systems. The VMAT1 gene maps to chromosome 8p21.3, a locus with strong evidence of linkage with schizophrenia. A recent study reported that a non-synonymous single nucleotide polymorphism (SNP) of the gene (Pro4Thr) was associated with schizophrenia. METHODS: We attempted to replicate this finding in a Japanese sample of 354 schizophrenics and 365 controls. In addition, we examined 3 other non-synonymous SNPs (Thr98Ser, Thr136Ile, and Val392Leu). Genotyping was performed by the TaqMan allelic discrimination assay. RESULTS: There was no significant difference in genotype or allele distribution of the three SNPs of Pro4Thr, Thr136Ile, or Val392Leu between patients and controls. There was, however, a significant difference in genotype and allele distributions for the Thr98Ser polymorphism between the two groups (P = 0.01 for genotype and allele). When sexes were examined separately, significant differences were observed in females (P = 0.006 for genotype, P = 0.003 for allele), but not in males. The Thr98 allele was more common in female patients than in female controls (odds ratio 1.69, 95% CI 1.19 2.40, P = 0.003). Haplotype-based analyses also provided evidence for a significant association in females. CONCLUSION: We failed to replicate the previously reported association of Pro4Thr of the VMAT1 gene with schizophrenia. However, we obtained evidence for a possible role of the Thr98Ser in giving susceptibility to schizophrenia in women. PMID- 17134515 TI - Prediction of protein submitochondria locations by hybridizing pseudo-amino acid composition with various physicochemical features of segmented sequence. AB - BACKGROUND: Knowing the submitochondria localization of a mitochondria protein is an important step to understand its function. We develop a method which is based on an extended version of pseudo-amino acid composition to predict the protein localization within mitochondria. This work goes one step further than predicting protein subcellular location. We also try to predict the membrane protein type for mitochondrial inner membrane proteins. RESULTS: By using leave-one-out cross validation, the prediction accuracy is 85.5% for inner membrane, 94.5% for matrix and 51.2% for outer membrane. The overall prediction accuracy for submitochondria location prediction is 85.2%. For proteins predicted to localize at inner membrane, the accuracy is 94.6% for membrane protein type prediction. CONCLUSION: Our method is an effective method for predicting protein submitochondria location. But even with our method or the methods at subcellular level, the prediction of protein submitochondria location is still a challenging problem. The online service SubMito is now available at: http://bioinfo.au.tsinghua.edu.cn/subMito. PMID- 17134516 TI - Can a disease-specific education program augment self-management skills and improve Health-Related Quality of Life in people with hip or knee osteoarthritis? AB - BACKGROUND: Patient education and self-management programs are offered in many countries to people with chronic conditions such as osteoarthritis (OA). The most well-known is the disease-specific Stanford Arthritis Self-Management Program (ASMP). While Australian and international clinical guidelines promote the concept of self-management for OA, there is currently little evidence to support the use of the ASMP. Several meta-analyses have reported that arthritis self management programs had minimal or no effect on reducing pain and disability. However, previous studies have had methodological shortcomings including the use of outcome measures which do not accurately reflect program goals. Additionally, limited cost-effectiveness analyses have been undertaken and the cost-utility of the program has not been explored. METHODS/DESIGN: This study is a randomised controlled trial to determine the efficacy (in terms of Health-Related Quality of Life and self-management skills) and cost-utility of a 6-week group-based Stanford ASMP for people with hip or knee OA. Six hundred participants referred to an orthopaedic surgeon or rheumatologist for hip or knee OA will be recruited from outpatient clinics at 2 public hospitals and community-based private practices within 2 private hospital settings in Victoria, Australia. Participants must be 18 years or over, fluent in English and able to attend ASMP sessions. Exclusion criteria include cognitive dysfunction, previous participation in self management programs and placement on a waiting list for joint replacement surgery or scheduled joint replacement.Eligible, consenting participants will be randomised to an intervention group (who receive the ASMP and an arthritis self management book) or a control group (who receive the book only). Follow-up will be at 6 weeks, 3 months and 12 months using standardised self-report measures. The primary outcome is Health-Related Quality of Life at 12 months, measured using the Assessment of Quality of Life instrument. Secondary outcome measures include the Health Education Impact Questionnaire, Western Ontario and McMaster Universities Osteoarthritis Index (pain subscale and total scores), Kessler Psychological Distress Scale and the Hip and Knee Multi-Attribute Priority Tool. Cost-utility analyses will be undertaken using administrative records and self report data. A subgroup of 100 participants will undergo qualitative interviews to explore the broader potential impacts of the ASMP. DISCUSSION: Using an innovative design combining both quantitative and qualitative components, this project will provide high quality data to facilitate evidence-based recommendations regarding the ASMP. PMID- 17134517 TI - Exploring the feasibility of a community-based strength training program for older people with depressive symptoms and its impact on depressive symptoms. AB - BACKGROUND: Depression is a disabling, prevalent condition. Physical activity programs may assist depression management in older people, ameliorate co-morbid conditions and reduce the need for antidepressants. The UPLIFT pilot study assessed the feasibility of older depressed people attending a community-based progressive resistance training (PRT) program. The study also aimed to determine whether PRT improves depressive status in older depressed patients. METHODS: A randomised controlled trial was conducted. People aged > or = 65 years with depressive symptoms were recruited via general practices. Following baseline assessment, subjects were randomly allocated to attend a local PRT program three times per week for 10 weeks or a brief advice control group. Follow-up assessment of depressive status, physical and psychological health, functional and quality of life status occurred post intervention and at six months. RESULTS: Three hundred and forty six people responded to the study invitation, of whom 22% had depressive symptoms (Geriatric Depression Scale, GDS-30 score > or = 11). Thirty two people entered the trial. There were no significant group differences on the GDS at follow-up. At six months there was a trend for the PRT intervention group to have lower GDS scores than the comparison group, but this finding did not reach significance (p = 0.08). More of the PRT group (57%) had a reduction in depressive symptoms post program, compared to 44% of the control group. It was not possible to discern which specific components of the program influenced its impact, but in post hoc analyses, improvement in depressive status appeared to be associated with the number of exercise sessions completed (r = -0.8, p < 0.01). CONCLUSION: The UPLIFT pilot study confirmed that older people with depression can be successfully recruited to a community based PRT program. The program can be offered by existing community-based facilities, enabling its ongoing implementation for the potential benefit of other older people. PMID- 17134519 TI - Disease management as an evolving role for respiratory therapists. PMID- 17134518 TI - In vitro antibacterial activity of some plant essential oils. AB - BACKGROUND: To evaluate the antibacterial activity of 21 plant essential oils against six bacterial species. METHODS: The selected essential oils were screened against four gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus vulgaris) and two gram-positive bacteria Bacillus subtilis and Staphylococcus aureus at four different concentrations (1:1, 1:5, 1:10 and 1:20) using disc diffusion method. The MIC of the active essential oils were tested using two fold agar dilution method at concentrations ranging from 0.2 to 25.6 mg/ml. RESULTS: Out of 21 essential oils tested, 19 oils showed antibacterial activity against one or more strains. Cinnamon, clove, geranium, lemon, lime, orange and rosemary oils exhibited significant inhibitory effect. Cinnamon oil showed promising inhibitory activity even at low concentration, whereas aniseed, eucalyptus and camphor oils were least active against the tested bacteria. In general, B. subtilis was the most susceptible. On the other hand, K. pneumoniae exhibited low degree of sensitivity. CONCLUSION: Majority of the oils showed antibacterial activity against the tested strains. However Cinnamon, clove and lime oils were found to be inhibiting both gram-positive and gram-negative bacteria. Cinnamon oil can be a good source of antibacterial agents. PMID- 17134520 TI - Mechanical ventilation in patients with Guillain-Barre syndrome. AB - BACKGROUND: Patients with Guillain-Barre syndrome are commonly exposed to prolonged mechanical ventilation. Specific data on ventilatory management of these patients have been limited. OBJECTIVE: To describe the practice of mechanical ventilation in patients with Guillain-Barre syndrome and evaluate risk factors for morbidity and mortality. METHODS: We describe a historical cohort of mechanically ventilated patients with Guillain-Barre syndrome in a tertiary-care center. We extracted database information on demographics, severity of illness, pulmonary function, and ventilatory management for the period 1976 to 1996. Primary outcomes were development of pulmonary complications, duration of ventilatory support, and mortality. RESULTS: Fifty-four patients met the inclusion criteria. After 1990, lower tidal volume (p = 0.031) and higher positive end-expiratory pressure (p = 0.003) were used than during the 1976 to 1990. Outcomes did not change significantly during the studied period. Forty-six patients (85%) survived to hospital discharge, and 39 (72%) were alive at 1-year follow-up. Ventilator-associated pneumonia was the most frequent complication (56%) and was associated with prolonged mechanical ventilation (p < 0.01). Atelectasis developed in 49%, and acute lung injury in 13%. All but 6 patients (89%) received tracheostomy. In 14 patients (30%) tracheostomy was placed > or = 14 days after intubation. When adjusted for atelectasis and severity of illness in a stepwise logistic regression analysis, delayed tracheostomy was associated with the development of ventilator-associated pneumonia (odds ratio 8.2, p = 0.029). CONCLUSIONS: Changes in ventilator practice did not affect outcomes of mechanically ventilated patients with Guillain-Barre syndrome. The majority of patients received tracheostomy, which should be considered early in the course of respiratory failure. PMID- 17134521 TI - Time to wean after tracheotomy differs among subgroups of critically ill patients: retrospective analysis in a mixed medical/surgical intensive care unit. AB - OBJECTIVE: To determine the time to wean from mechanical ventilation and time spent off the ventilator per day after tracheotomy in critically ill patients in a 28-bed mixed medical and surgical intensive care unit (ICU) in Amsterdam, Netherlands. METHODS: We conducted a retrospective analysis of consecutive patients during the 14-month period from November 1, 2003, through January 1, 2005. Included were translaryngeally intubated mechanically ventilated patients who received a tracheotomy during their ICU stay. RESULTS: Of all the patients admitted to the ICU, 129 (7%) received a tracheotomy. Significantly more tracheotomies were performed in neurosurgery/neurology patients and in those admitted for acute conditions (16% and 12%, respectively). Tracheotomy was performed a median 8 days (interquartile range 4-13 d) after ICU admission. For all the patients, the median time to wean after tracheotomy was 5 days (interquartile range 2-11 d). Neurosurgery/neurology patients and patients in the cardiology subgroup needed significantly less time to wean from mechanical ventilation than did patients in other subgroups (3 d, interquartile range 2-7 d, and 3 d, interquartile range 2-5 d, respectively, p < 0.05). There was a significant association between admission group and neurological status at the time of tracheotomy. A low Glasgow coma scale score was associated with shorter time to wean. Within 1 week after tracheotomy, the probability of the patient having breathed spontaneously, without ventilator assistance, for > 4 h/d was 89%, 78% for > 8 h/d, and 72% for > 12 h/d. By day 28, the probability of the patient having breathed spontaneously for > 4 h/d was 98%, 97% for > 8 h/d, and 94% for > 12 h/d. CONCLUSION: Time to wean from after tracheotomy differed among the subgroups in our ICU. After tracheotomy, the majority of patients were quickly able to breathe spontaneously without assistance of the mechanical ventilator for several hours per day. Patients who require tracheotomy only for airway protection wean sooner than other patients. PMID- 17134522 TI - Twenty-four-hour ambulatory oximetry monitoring in COPD patients with moderate hypoxemia. AB - BACKGROUND: The relationship of diurnal desaturations (oximetry-measured blood oxygen saturation [S(pO(2))] < 90%) during activities of daily living and clinical aspects in patients with chronic obstructive pulmonary disease (COPD) and moderate hypoxemia has not been systematically evaluated. METHODS: We prospectively studied 88 patients with stable COPD (forced expiratory volume in the first second [FEV(1)] < 80% of predicted, ratio of FEV(1) to forced vital capacity < 70% of predicted, and P(aO(2)) 60-70 mm Hg) with 24 hours of ambulatory oximetry. Desaturators were defined as those who spent >/= 30% of the time with S(pO(2)) < 90%. Patients engaged in their usual activities of daily living. We correlated these desaturations with the following variables, measured immediately before the 24 hours of oximetry: body mass index, dyspnea (measured with the modified Medical Research Council dyspnea scale), gas exchange, pulmonary function, quality of life (measured with the Saint George's respiratory questionnaire), and comorbidity (measured with the Charlson index). RESULTS: Thirty-three (38%) of the patients were desaturators: 50% nocturnal and 22% diurnal. We also measured daytime arterial blood gas values from arterial blood samples and found that the desaturators had higher P(aCO(2)) (p = 0.001) and lower P(aO(2)) (p = 0.007) than the nondesaturators. There were no differences in the other variables. The correlation between nocturnal and diurnal time with S(pO(2)) < 90% was r(2) = 0.67, and the concordance was low (Cohen's kappa 0.43, p < 0.001). CONCLUSIONS: Patients with stable COPD and moderate hypoxemia have frequent and potentially important desaturations during activities of daily living and at night. In addition, there is a big difference in the profile and degree of nocturnal and diurnal desaturations. Twenty-four hours of oximetry provides valuable information for comprehensive evaluation of patients with COPD. PMID- 17134523 TI - Evaluation of an oxygen protocol in long-term care. AB - OBJECTIVE: The purpose of this study was to determine the effect of protocol directed recommendations for oxygen therapy implemented by respiratory therapists in skilled nursing facilities. We hypothesized that the use of an oxygen protocol would reduce the number of missing and incomplete orders and unnecessary oxygen use in skilled nursing facilities. METHODS: We studied patients who required oxygen therapy in 17 Ohio-based skilled nursing facilities. Respiratory therapists assessed the need for oxygen therapy. Recommendations for oxygen use and orders were made in accordance with an algorithm-based protocol and guidelines established by the Ohio Department of Health. Data were prospectively collected from January 1 through March 31, 2005. RESULTS: Of 346 eligible patients, 261 had complete data and comprised the study sample. The mean +/- SD age was 83 +/- 11.8 years, and 79% were male. Payer mix included Medicaid (46%), Medicare Part A (36%), private pay (11%), and hospice (7%). Orders for oxygen therapy were incomplete or missing in 18% of the population. A total of 1,175 billed days were saved, which corresponded to a cost savings of US dollars 6,768. CONCLUSIONS: Successful implementation of an oxygen protocol can improve compliance with accreditation agency requirements by reducing the number of missing and/or incomplete orders for oxygen therapy. Financial and patient outcomes can also be enhanced by discontinuing unnecessary oxygen use and initiating oxygen therapy when clinically needed. PMID- 17134524 TI - Compliance of physicians with documentation of an asthma management protocol. AB - BACKGROUND: Studies continue to show poor physician compliance with asthma management guidelines in clinical practice. However, standardized protocols specifically designed to be practical and user-friendly improve patient outcomes. OBJECTIVE: To determine the degree of physicians' compliance with the documentation of an asthma management protocol in a university hospital. METHODS: A simple asthma management protocol was designed and applied in our pulmonary clinic and primary care clinic for asthma. The protocol was based on the 1998 Manual for the Management of Asthma, from the Oman Ministry of Health, which follows internationally recognized guidelines. The protocol consisted of 4 sections: clinical history, peak expiratory flow (PEF) data, medication section, and simplified asthma management guidelines. RESULTS: All 30 physicians scheduled to conduct asthma clinics in the pulmonary clinic (14 physicians) and the primary care clinic (16 physicians) agreed to use the protocol. A total of 282 protocol forms were collected: 130 forms from 6 senior physicians and 152 from 24 junior physicians. Documentation of the entire clinical history was 65%, with the senior physicians scoring significantly higher documentation-completion rates (82%) for all components of the history than the junior physicians (50%). Documentation of all PEF data was poor (26%), despite high documentation of the PEF value itself (95%). There were significant differences in documentation of percent-of predicted PEF between junior physicians in primary care clinic (70%) and other physicians (19%). Documentation of the entire medication section was only 34%. Although documentation of prescribed medicines was high (92%), compliance (48%) and inhaler technique (49%) documentation was low, with similar patterns demonstrated by all physicians. Documentation of the entire protocol by all physicians was low (9%), with junior physicians in the primary care clinic completing 28% of their forms. CONCLUSIONS: Our protocol enabled us to identify opportunities for improvement in documentation of asthma management in both the pulmonary and primary care clinics. The findings highlight the need for regular asthma education programs for all physicians, with a focus on documentation of performance skills such as monitoring of PEF and inhaler technique. PMID- 17134525 TI - Adolescents with asthma: learning needs and internet use assessment. AB - OBJECTIVE: To identify particular learning needs among adolescents with asthma and explore the potential utility of the Internet in addressing adolescents' expressed learning needs. METHODS: In 2004 and 2005, 6 gender-specific and age specific focus groups were conducted among adolescents, ages 12-18 years, with mild intermittent to severe persistent asthma. Thematic analysis of participants' statements from transcribed group sessions were conducted by the research team, who compared field notes, reviewed focus group transcripts, and validated emerging and final themes. RESULTS: The majority of participants denied previous experience with formal asthma education. Participants reported their limited knowledge of asthma and its management. Health-care providers were recognized as the most credible source of asthma information. Compared to the older adolescents, the younger adolescents expressed stronger motivation to learn about asthma. The participants identified asthma learning needs for others, including peers, teachers, and parents. The importance of socialization, support, and information-sharing with other adolescents with asthma was perceived by participants in all age and gender groups. Participants discussed their Internet use and articulated suggestions regarding potential uses of the Internet to assist in adolescent asthma management. CONCLUSIONS: This study underscores the necessity of an early intervention to take advantage of younger adolescents' greater interest in learning about asthma, and highlights the importance of incorporating peer dynamics in designing interventions for adolescents. Our findings also illustrate the variety of ways that the Internet may be useful in developing a technology-driven intervention for adolescents. PMID- 17134526 TI - Comparison of published pressure gradient symbols and equations in mechanics of breathing. AB - In the literature of pulmonary medicine we found dismaying diversity of and inconsistency in terms used to describe physiologic pressure gradients. Standardized terms, definitions, symbols, and equations published by the American Physiological Society, the American College of Chest Physicians, the American Thoracic Society, and the American Association for Respiratory Care have not been consistently used. Rather, researchers have often used their own definitions for transpulmonary pressure, transairway pressure, transthoracic pressure, transrespiratory pressure, and transdiaphragmatic pressure. We describe the variety of definitions and equations we found for those terms. We contend that it would benefit researchers, students, clinicians, and educators to define these terms precisely and use them consistently. PMID- 17134527 TI - Zen and the art of nomenclature maintenance: a revised approach to respiratory symbols and terminology. AB - In this paper we point out that there are different entities involved in the mathematical descriptions, or models, of the respiratory system: variables and parameters. These, in turn, can be divided into different types. Variables can be primary variables, difference variables, or change variables. Difference variables express the difference between primary variables measured simultaneously at 2 locations. Change variables are primary variables measured relative to fixed reference values. Parameters that appear in input-output models that are valid over a wide range of inputs can be interpreted as properties. There are 3 levels of properties, depending on the detail included in the model. If the model specifically includes the geometry of the system and the substances of which the system is composed, then the parameters in the model are material properties. If the model includes the general structures that compose the system, the parameters are structural properties. And if the model describes the behavior of the system as a whole with no detail included pertaining to internal makeup, then the parameters can be considered system properties. Parameters that appear in mathematical descriptions of input and/or output wave shapes can be interpreted as waveform characteristics. General waveform characteristics are attributes of arbitrary inputs and/or outputs. However, in those special cases in which a system is subjected to a well-defined, specialized input and the output waveform is described mathematically (even if only at a single point), the parameter(s) in such descriptions can be considered system response characteristic(s). We suggest that the symbols and names given to these various entities should follow well-defined guidelines that distinguish among the entity types. These guidelines should include symbol and name conventions and also sign conventions and expected unit ranges on appropriate measurement scales. One such set of conventions would be as follows. Italicize all variables. Use upper-case for primary (absolute) variables. Use the delta symbol (Delta) to denote difference variables (difference between 2 locations). Use lower-case letters for change variables (change relative to a reference, or operating, point) and for abbreviations (eg, "pl" for "pleural"). Use upper-case characters to represent the initial letters of words (eg, "AO" for "airway opening"). Make bold nonitalicized groups of letters used for properties (upper-case, lower-case, multi-height). Do not bold or italicize groups of letters used for characteristics (upper-case, lower-case, multi-height). Compound symbols are those that include subscripts and/or superscripts. Subscripts following a symbol indicate location, direction, or index (time); if more than one subscript, separate them by commas. Superscripts following a symbol indicate a component, or it can indicate a power if the symbol is enclosed in parentheses. Letters on the same line as initial letter but in smaller type are part of the generic symbol. Arguments are enclosed in parentheses; parentheses are also used to isolate compound symbols from powers or additional subscripts. Adapt currently standard symbols to retain their identity but conform to the above conventions. The sign of an entity is always dictated by and its interpretation is based on the model in which the entity is used. Units used are consistent with measurement resolution and accuracy. Copious examples of the applications of this set of suggested conventions are given in the text and in 4 tables. Our hope is that the presentation of these suggestions will start a dialogue in the field and will influence journal and book publishers to adopt a consistent set of conventions for the names and symbols used for respiratory-system-related terms. PMID- 17134528 TI - Effect of early rearing conditions on alcohol drinking and 5-HT1A receptor function in C57BL/6J mice. AB - We have evaluated in C57BL/6J mice the effect of maternal separation and post weaning social isolation on ethanol intake, and on serotonin1A (5-HT1A) receptor function at the level of receptor-G protein interaction in the hippocampus and dorsal raphe nucleus. From postnatal days 2-14, litters were separated from the mother for 15 min (Handled) or for 180 min (Maternal separation). After weaning, pups were housed in pairs or in social isolation. At 2 months of age, ethanol intake and preference in mice were assessed using the two-bottle choice paradigm. Maternal separation increased ethanol preference in female mice that were subsequently housed in isolation. By contrast, post-weaning isolation increased ethanol preference and consumption in male mice regardless of pre-weaning rearing conditions. The increased ethanol preference and intake were limited to a 5% (v/v) concentration of ethanol. Our data suggest that adolescent mice are susceptible to the effects of post-weaning social isolation as shown by increased ethanol preference and consumption. Using quantitative autoradiography, 5-HT1A receptor number and function were determined by the binding of [3H]WAY-100635, and by [35S]GTPgammaS binding stimulated by the 5-HT1A receptor agonist 8-OH DPAT, respectively. The binding experiments were done at approximately 3 months after the end of the two-bottle choice test in an attempt to minimize direct effects of ethanol drinking on 5-HT1A receptor function and number. 5-HT1A receptor-stimulated [35S]GTPgammaS binding in the dorsal raphe nucleus was increased in animals reared after weaning in isolation vs. in pairs, regardless of gender or pre-weaning rearing conditions. Our data suggest that there are long term neurochemical consequences of social isolation of adolescent mice, specifically increased 5-HT1A receptor function in the dorsal raphe nucleus. PMID- 17134529 TI - Molecular mimicry and horror autotoxicus: do chlamydial infections elicit autoimmunity? AB - All species of the order Chlamydiales are obligate intracellular eubacterial pathogens of their various hosts. Two chlamydial species, Chlamydia trachomatis and Chlamydia pneumoniae, are primarily human pathogens, and each is known to cause important diseases. Some strains of C. trachomatis are sexually transmitted and frequently cause severe reproductive problems, primarily in women. Other strains of the organism serve as the aetiological agents for blinding trachoma, still the leading cause of preventable blindness in underdeveloped nations. C. pneumoniae is a respiratory pathogen known to cause community-acquired pneumonia. Importantly, both organisms engender an immunopathogenic response in the human host, and both have been associated with widely diverse, relatively common and currently idiopathic chronic diseases, most of which include an important autoimmune component. In this article, we explore the available experimental data regarding the possible elicitation of autoimmunity in various contexts by chlamydial infection, and we suggest several avenues for research to explore this potentially important issue further. PMID- 17134530 TI - A multi-centre prospective case-control study of campylobacter infection in persons aged 5 years and older in Australia. AB - There are an estimated 277,000 cases of campylobacteriosis in Australia each year, most of which are thought to be sporadically acquired. To explore causes for these infections, we conducted a multi-centre case-control study of patients and community controls across five Australian States during 2001-2002. A total of 881 campylobacter cases and 833 controls aged 5 years were recruited into the study. Crude logistic analyses were conducted within various food and non-food exposure groups. A final most parsimonious multivariable logistic regression model was developed and adjusted odds ratios (aOR), 95% confidence intervals (95% CI) were derived together with adjusted population attributable risks (PAR). Consumption of undercooked chicken (aOR 4.7, 95% CI 2.6-8.4) and offal (aOR 2.0, 95% CI 1.0-4.0), ownership of domestic chickens aged<6 months (aOR 12.4, 95% CI 2.6-59.3) and domestic dogs aged<6 months (aOR 2.1, 95% CI 1.1-4.2) were found to be independent risk factors for illness in the final model. The PAR proportions indicate that eating chicken meat, either cooked or undercooked may account for approximately 30% of campylobacter cases that occur each year in Australia. These results justify the continued need for education of consumers and foodhandlers about the risks associated with the handling of raw chicken and the potential for cross-contamination. PMID- 17134531 TI - Survey on the occurrence of Brachyspira species and Lawsonia intracellularis in children living on pig farms. AB - The occurrence of Brachyspira species and Lawsonia intracellularis was investigated by PCR analyses of faeces from 60 children living on European pig farms. In addition, 60 other children were included as controls. Two samples were positive for B. aalborgi but B. pilosicoli and L. intracellularis were not demonstrated. PMID- 17134532 TI - Genetic and environmental influences on dimensional representations of DSM-IV cluster C personality disorders: a population-based multivariate twin study. AB - BACKGROUND: The DSM-IV cluster C Axis II disorders include avoidant (AVPD), dependent (DEPD) and obsessive-compulsive (OCPD) personality disorders. We aimed to estimate the genetic and environmental influences on dimensional representations of these disorders and examine the validity of the cluster C construct by determining to what extent common familial factors influence the individual PDs. METHOD: PDs were assessed using the Structured Interview for DSM IV Personality (SIDP-IV) in a sample of 1386 young adult twin pairs from the Norwegian Institute of Public Health Twin Panel (NIPHTP). A single-factor independent pathway multivariate model was applied to the number of endorsed criteria for the three cluster C disorders, using the statistical modeling program Mx. RESULTS: The best-fitting model included genetic and unique environmental factors only, and equated parameters for males and females. Heritability ranged from 27% to 35%. The proportion of genetic variance explained by a common factor was 83, 48 and 15% respectively for AVPD, DEPD and OCPD. Common genetic and environmental factors accounted for 54% and 64% respectively of the variance in AVPD and DEPD but only 11% of the variance in OCPD. CONCLUSION: Cluster C PDs are moderately heritable. No evidence was found for shared environmental or sex effects. Common genetic and individual environmental factors account for a substantial proportion of the variance in AVPD and DEPD. However, OCPD appears to be largely etiologically distinct from the other two PDs. The results do not support the validity of the DSM-IV cluster C construct in its present form. PMID- 17134533 TI - Rhinocerebral mucormycosis with orbital and intracranial extension: a case report and review of optimum management. AB - Rhinocerebral mucormycosis is a devastating, rapidly progressive and often fatal opportunistic fungal infection predominantly affecting individuals with underlying metabolic and/or immunological compromise. Intracranial extension of the disease has invariably been associated with mortality.We present a review of optimum management of rhinocerebral mucormycosis and a case report of sinonasal mucormycosis with intracranial and orbital extension which was treated successfully with a combination of systemic liposomal amphotericin B therapy and wide surgical debridement. PMID- 17134534 TI - Topical antibiotic ototoxicity: does it influence our practice? AB - INTRODUCTION: We hypothesised that general practitioners and ENT specialists manage discharging ears differently. This study was designed to investigate this further. METHODS AND MATERIALS: Postal questionnaires were sent to all general practitioners in the Birmingham area and all UK consultants on the British Association of Otolaryngology-Head and Neck Surgery address list. RESULTS AND DISCUSSION: In the presence of an intact tympanic membrane, 99 per cent of consultants and 90 per cent of general practitioners would use topical antibiotics. In the presence of a perforated tympanic membrane, 97 per cent of consultants would continue to use topical antibiotics, compared with only 43 per cent of general practitioners. This was attributed to a fear of ototoxicity. If a topical non-ototoxic antibiotic of proven efficacy could be made available, 93 per cent of consultants and 88 per cent of general practitioners in this study would seriously consider using it as first line treatment. CONCLUSION: The majority of general practitioners would not use topical antibiotics in the presence of a perforated tympanic membrane. Most doctors would consider using a non-ototoxic topical antibiotic as first line treatment should one be made available. PMID- 17134535 TI - Sphenochoanal polyp with heterotopic glial tissue. AB - An isolated polyp arising from the sphenoid sinus is rare. Due to the presence of important structures adjacent to the sphenoid sinus, a proper pre-operative radiological and nasal endoscopic evaluation is mandatory, along with a neurological and ophthalmological assessment. We report the case of a 23-year-old man with a sphenochoanal polyp with heterotopic glial tissue in the stroma. A Medline search did not reveal any previously reported cases of glial tissue in the sphenoid sinus presenting as a sphenochoanal polyp. PMID- 17134536 TI - Ossifying fibroma of the mandible with primary hyperparathyroidism due to non familial parathyroid adenoma. AB - Ossifying fibroma is an uncommon benign osteogenic neoplasm arising from cells of the periodontal ligament, typically with a slowly progressive enlargement of the affected bone. The neoplasm sometimes presents with hyperparathyroidism, most of which cases are due to familial parathyroid tumours. We report a rare case of ossifying fibroma of the mandible which showed very rapid growth and presented with primary hyperparathyroidism due to non-familial parathyroid adenoma. Despite improvement of parathyroid dysfunction after removal of the parathyroid adenoma, the tumour continued to grow very aggressively. The case required partial mandibular resection for complete resection of the tumour, and fixation of the remaining mandible with a titanium plate. PMID- 17134537 TI - Diagnostic adequacy and accuracy of fine needle aspiration cytology in neck lump assessment: results from a regional cancer network over a one year period. AB - AIM: To establish the diagnostic accuracy and adequacy of fine needle aspiration cytology (FNAC) within a regional cancer network, and to determine what service improvements may be required to allow successful implementation of an FNAC-based, 'one-stop' head and neck clinic, as proposed by the current National Institute for Clinical Excellence guidelines. MATERIALS AND METHODS: The Sussex cancer network serves a population of 1,200,000 and contains five hospitals within three acute trusts. In 2004, an audit was undertaken retrospectively to examine the diagnostic adequacy and accuracy of head and neck FNAC across the network. Comparisons were then made with the results of subsequent relevant surgery. For the purposes of the audit, FNAC was subdivided into three main groups: salivary gland, thyroid gland and neck node. As part of the data analysis, we also noted the clinical source of the FNAC and whether it was performed blind or under image guidance. RESULTS: In 2004, 712 FNAC procedures were undertaken in 647 patients, 276 of whom underwent subsequent surgery. Fine needle aspiration cytology was non diagnostic in 52 per cent of patients in the neck node group, in 50 per cent in the salivary gland group and in 30 per cent in the thyroid group. With these non diagnostic results removed, statistical analysis was performed on data from those patients who had undergone both FNAC and subsequent surgery. This gave a sensitivity of 89 per cent and a specificity of 57 per cent in the neck node group, a sensitivity of 64 per cent and specificity of 100 per cent in the salivary gland group, and a sensitivity of 62 per cent and specificity of 86 per cent in the thyroid group. Diagnostic problems with FNAC were noted, particularly in the differentiation of reactive nodal hyperplasia from lymphoma and in diagnosing follicular thyroid lesions. Ultrasound guidance was used in 50 per cent of the thyroid FNAC procedures but in only a minority of patients in the neck node and salivary gland groups. CONCLUSION: This audit demonstrated widespread diagnostic difficulties associated with head and neck FNAC in a large patient sample. It is likely that these problems will be mirrored in other cancer networks. In order for one-stop head and neck clinics to succeed, the non diagnostic rate of FNAC in particular must be minimised. There are strategies to enable this, depending on local resources, including increased access to cytologists or cytology technicians, diagnostic ultrasound, image guidance for FNAC and the use of ultrasound-guided core biopsy. PMID- 17134538 TI - Understanding submucosal electrosurgery for the treatment of nasal turbinate enlargement. AB - The surgical management of inferior turbinate enlargement is controversial. Submucosal electrosurgical techniques for turbinate reduction include conventional diathermy, radiofrequency tissue reduction and coblation. All electrosurgical techniques use radiofrequency electricity to damage turbinate tissue but differ in the control and delivery of energy. This review will examine the history of submucosal electrosurgery and clarify the various techniques. This review will also examine the evidence for the efficacy and safety of electrosurgery for the treatment of nasal turbinate enlargement, and will make a case that no progress will be made in clinical trials on the safety and efficacy unless there is standardisation of equipment and techniques in nasal electrosurgery. PMID- 17134539 TI - Lack of ghrelin secretion in response to fasting in cholecystokinin-A (-1), -B ( 2) receptor-deficient mice. AB - Cholecystokinin receptors (CCK-Rs) have been classified into two subtypes: CCK-AR (1R) and -BR (2R). We generated CCK-AR(-/-), CCK-BR(-/-), and CCK-AR(-/-)BR(-/-) mice and found that the gastric emptying of a liquid meal was increased in CCK BR(-/-) and AR(-/-)BR(-/-) mice, compared with wild-type and CCK-AR(-/-) mice. Given that enhanced gastric emptying leads to eating, food intake after overnight fasting was examined, as was the effect of CCK-8S on food intake. Male mice 6-8 months of age were deprived of food for 16 h with free access to water, after which they were injected intraperitoneally (0.1 ml/mouse) with either vehicle or CCK-8 (0.3, 1.0, or 3.0 nmol/mouse), and their food intake was monitored for 4 h. CCK-8S inhibited food intake in wild-type and CCK-BR(-/-) mice, but not in CCK AR(-/-) or AR(-/-)BR(-/-) mice. Unexpectedly, we observed a lower food intake in CCK-AR(-/-)BR (-/-) mice treated with vehicle than in mice of the other genotypes. To examine the mechanism of decrease in food intake in CCK-AR(-/-)BR( /-) mice, the involvement of ghrelin was determined in wild-type and CCK-AR(-/ )BR(-/-) mice. Fasting plasma ghrelin levels were significantly lower in CCK-AR ( /-)BR(-/-) mice than in wild-type mice, and no increase in response to fasting was observed in CCK-AR(-/-)BR(-/-) mice. An administration of acyl-ghrelin produced a small increase in food intake in CCK-AR(-/-)BR(-/-) mice, but not to the levels of wild-type mice. In conclusion, CCK-AR(-/-)BR(-/-) mice showed lower food intake as well as lower response to exogenous ghrelin, and a lower plasma ghrelin level after fasting, though which receptor is more important is unknown. PMID- 17134540 TI - Functional residual capacity and airway resistance of the rat measured with a heat- and temperature-adjusted body plethysmograph. AB - The functional residual capacity (FRC) and airway resistance (R(aw)) of the rat were measured, using a newly designed body plethysmograph (BPG), the inner environment of which was maintained at body temperature and was water-vapor saturated. The subjects were anesthetized and tracheally intubated male Wistar rats (n = 15). After measuring the FRC and R(aw), we analyzed the effects of inhaled methacholine (Mch, 0-8 mg/ml) on R(aw).The determined FRC was 5.37 +/- 0.22 ml (mean +/- SE). An almost linear relationship between box pressure and respiratory flow was obtained when the difference between box-gas temperature and the rectal temperature of the rat was less than 1.0 degrees C. The R(aw) at FRC was 0.230 +/- 0.017 cm H(2)O/ml/s. It increased proportionally with increases in the Mch concentration. When the dynamic changes in R(aw) were analyzed, the R(aw) was found to progressively increase during expiration; this increase continued throughout inspiration. Thus in the rat, R(aw) is not simply a function of changes in lung volume. In conclusion, the humidity- and temperature-adjusted BPG provided an absolute and possibly dynamic value of R(aw). PMID- 17134541 TI - [Roles of immunopathological technique in diagnosis of renal diseases]. PMID- 17134542 TI - [Pathology of kidney tumors: introduction of the 2004 WHO classification]. PMID- 17134543 TI - [Primary adenosquamous cancer of cecum: report of a case]. PMID- 17134544 TI - [Papillary urothelial neoplasms of low malignant potential]. PMID- 17134545 TI - [Relationship between chromosome 8 alterations and Gleason score in prostatic adenocarcinoma]. AB - OBJECTIVE: To study the gain of chromosome 8 and c-myc gene and lipoprotein lipase gene status in prostatic adenocarcinoma of Chinese patients, and to analyze the relationship between chromosome 8 alterations and Gleason score of prostatic cancer. METHODS: Formalin-fixed, paraffin-embedded prostatic biopsy tissues from 34 Chinese patients with untreated prostatic adenocarcinoma were studied by three-color fluorescence in situ hybridization (FISH) using ProVysion(TM) probe kit. The materials included 1 case with Gleason score 5, 10 cases with Gleason score 6, 14 cases with Gleason score 7, 4 cases with Gleason score 8 and 5 cases with Gleason score 9. The relationship between Gleason score and chromosome 8 aneusomy, c-myc and lipoprotein lipase gene copy number, including gene amplification or deletion, were analyzed. RESULTS: Seventeen (50%) of the 34 cases studied had gain of chromosome 8, while 21 cases (61.8%) had gain of c-myc gene copy number, 15 cases (44.1%) had lipoprotein lipase gene monosomy, 23 cases (67.6%) had c-myc gene amplification, 21 cases (61.8%) had deletion of lipoprotein lipase gene and 16 cases (47.1%) had lipoprotein lipase gene deletion coupled with c-myc gene amplification. In general, at least one type of chromosome 8 alteration was identified in 85.3% of cases (29/34). Gain of chromosome 8 was strong significant associated with Gleason score (P = 0.0006). A positive correlation between increased c-myc copy number and high Gleason score was also noted (P = 0.0035). On the other hand, loss of lipoprotein lipase gene was negatively correlated with high Gleason score (P = 0.0383). In addition, the association of c-myc gene amplification with high Gleason score was noted after age adjustment (P = 0.0462). CONCLUSIONS: Alterations in chromosome 8 are common in prostatic adenocarcinoma occurring in Chinese patients. There is a correlation between Gleason score and gain of chromosome 8, increased c-myc gene copy number, c-myc gene amplification and lipoprotein lipase gene deletion. C-myc gene amplification accompanied by lipoprotein lipase gene deletion is also a common occurrence in prostatic cancer. Our data suggest that chromosome 8 alterations may play some roles in the development and progression of prostatic adenocarcinoma. PMID- 17134546 TI - [Diffuse large B-cell lymphoma with expression of anaplastic lymphoma kinase protein: clinicopathologic and immunohistochemical study of 5 cases]. AB - OBJECTIVE: To study the clinicopathologic features of diffuse large B-cell lymphoma (DLBCL) with expression of anaplastic lymphoma kinase (ALK) protein. METHODS: Nine hundred and forty-five (945) cases of DLBCL (including 177 consultation cases) diagnosed according to the 2001 World Health Organization classification of tumors of hematopoietic and lymphoid tissues were enrolled into the study. Immunohistochemical study for anti-ALK-11 was performed using LSAB technique. The ALK-positive cases were further confirmed by immunohistochemical study using EnVision technique. Only ALK-positive cases by EnVision technique were further analyzed by immunostaining for antigens including CD20, CD3, CD30, EMA, granzyme-B, TIA-1 and PC. Immunoglobulin heavy chain gene rearrangement study was also performed and follow-up data collected. RESULTS: There were altogether 5 (4 males and 1 female) cases of DLBCL showing expression of ALK protein. The age of the patients ranged from 34 to 72 years. All were primary nodal DLBCL. One case belonged to clinical stage I, 2 in stage II and 2 in stage III. The duration of follow up ranged from 4 to 32 months. Three patients subsequently died and the longest survival was 32 months. Morphologic subtypes included centroblastic 2, anaplastic 1, immunoblastic with plasmacytoid differentiation 1 and plasmablastic 1. Immunohistochemically, 4 cases were CD20 positive (including 2 centroblastic, 1 anaplastic and 1 immunoblastic cases). The plasmablastic case expressed kappa light chain and was negative for CD20. Rearrangement of immunoglobulin heavy chain gene was demonstrated in all 5 cases studied. As for ALK protein staining, a mixed membranous and cytoplasmic (1 immunoblastic case), granular cytoplasmic (2 centroblastic and 1 anaplastic cases) and mixed nuclear and cytoplasmic (1 plasmablastic case) patterns were observed. CONCLUSIONS: Expression of ALK protein is a rare phenomenon in DLBCL and can be seen in centroblastic, anaplastic, immunoblastic and plasmablastic subtypes. It is often associated with aggressive clinical behavior and worse prognosis. A new pattern of ALK protein expression, mixed membranous and cytoplasmic, is reported. PMID- 17134547 TI - [Real-time PCR analysis of beta-catenin mRNA in sporadic colorectal cancers]. AB - OBJECTIVE: To detect beta-catenin mRNA levels in sporadic colorectal cancers (SCRC) and adjacent normal colorectal mucosa, and to investigate the association between the beta-catenin mRNA level and its aberrant expression and clinicopathological parameters. METHODS: The concentration of beta-catenin mRNA in 81 SCRCs and 28 adjacent normal colorectal mucosa specimens was determined by TaqMan real-time quantitative RT-PCR. The ratio of beta-catenin cDNA copies/GAPDH cDNA copies was used to represent the mRNA expression level in different tissues. The beta-catenin protein expression was determined by the EnVision two-step immunohistochemical method. RESULTS: beta-catenin mRNA levels in SCRCs (2.527 +/- 2.284) were lower than those in the adjacent normal colorectal mucosa (5.003 +/- 3.326), P < 0.05. In addition, beta-catenin mRNA levels in lymph node-positive cases and tumors with ulcerative and infiltrating growth types were significantly lower (1.827 +/- 1.288, 2.202 +/- 2.035) than those in lymph node-negative cases and polypoid growth type tumors (3.359 +/- 2.881, 3.108 +/- 2.610), P < 0.05. No significant difference of beta-catenin mRNA level was found between cases with aberrant beta-catenin cytoplasm or nuclear expression and those without. CONCLUSIONS: SCRCs express lower levels of beta-catenin mRNA than normal colorectal mucosa. Such lower level expression is associated with lymph node metastasis and tumors with ulcerative and infiltrative growth pattern. Aberrant cytoplasmic and nuclear expression of beta-catenin appears unrelated to the lower mRNA levels. Quantitative detection of beta-catenin mRNA may be a useful approach to monitor the biological behavior of SCRCs. PMID- 17134548 TI - [TPX2 expression and its significance in squamous cell carcinoma of lung]. AB - OBJECTIVE: To study the expression of targeting protein for Xklp2 (TPX2) and its significance in squamous cell carcinoma (SCC) of the lung. METHOD: Two SCC cell lines and 4 immortalized bronchial epithelial cell lines (as a precancerous model) were examined by Western blot for TPX2 expression. Reverse transcription polymerase chain reaction analysis for TPX2 was also performed using tumor tissues from 21 patients with SCC of the lung. The expression of TPX2 was studied by immunohistochemistry (using tissue microarray) on paraffin-embedded sections of pulmonary SCC and corresponding precancerous lesions from a group of 319 patients. RESULTS: TPX2 was variably expressed in all the cell lines studied. Compared with matched controls using normal lung tissue, high level of TPX2 mRNA was detected in 16 of the 21 SCC tumor tissue samples analyzed. Immunohistochemical study showed that TPX2 was mainly present in tumor tissues but not in normal controls. The expression of TPX2 correlated with tumor grade, stage and nodal status. As for precancerous lesions, the level of TPX2 was also increased, in accordance with the degree of dysplasia. CONCLUSIONS: Expression of TPX2 may play a role in carcinogenesis of bronchial epithelium and tumor progression of pulmonary SCC. It may also represent a potential biomarker for surveillance of SCC of lung. PMID- 17134549 TI - [Pathologic changes caused by highly pathogenic H5N1 avian influenza virus: postmortem study of a case]. AB - OBJECTIVE: To report the postmortem findings of a case of highly pathogenic H5N1 avian influenza virus occurring in human beings. METHODS: Postmortem examination was carried out in a deceased caused by highly pathogenic H5N1 avian influenza virus. Detailed light microscopy of major organs, including heart, lungs, liver, spleen, kidneys and brain, was performed. The lung tissue was further investigated by histochemistry, immunohistochemistry and electron microscopy. RESULTS: Major histopathologic changes in lungs secondary to highly pathogenic H5N1 avian influenza virus included diffuse alveolar damage, hyaline membrane formation and focal hemorrhage. Some of the alveolar spaces contained lightly eosinophilic liquid, lymphocytes, macrophages, plasma cells and small number of neutrophils. Congested capillaries were commonly seen in the alveolar septa which were focally rimmed by hyaline membrane. Immunohistochemical study showed that the lymphocytes were mainly of T lineage and macrophages were also demonstrated. CONCLUSIONS: Highly pathogenic H5N1 avian influenza virus causes pathologic changes mostly in lungs, including diffuse alveolar damage and acute exudative changes (involving mainly T lymphocytes and macrophages). The resulting parenchymal destruction, consolidation, pulmonary edema and hemorrhage eventually lead to respiratory distress and death. PMID- 17134550 TI - [Biological implications of the inhibition of survivin by RNA interference in human androgen-independent prostate carcinoma with highly metastatic potential]. AB - OBJECTIVE: To determine the expression level of survivin in androgen-independent prostate carcinoma, and to investigate the biological role of survivin in invasion and metastasis of androgen-independent prostate carcinoma. METHODS: Highly metastatic prostatic cancer cell line PC-3M-1E8 was stably transfected with pSilencer plasmid targeting survivin expression by RNA interference. The biological effects were observed, including anchorage-independent growth, in vitro invasion by soft agar colony formation and Boyden chamber assay, and also in vivo tumorigenesis in nude mice. Cell cycle and apoptosis indices were evaluated by flow cytometry and Western blot analysis of bioactive fragments of caspase 3. RESULTS: The expression of survivin in transfected PC-3M-1E8 cells was markedly depressed at both mRNA and protein levels (about 78% to 80%) as compared with control. The growth of tumor cells was retarded by anchorage-independent growth assay. The survivin transfectants formed smaller and fewer colonies (14.33 +/- 3.51) than the negative (52.33 +/- 6.81) and blank controls (54.00 +/- 6.00). Inhibition of survivin expression was correlated with enhanced apoptosis of tumor cells (percentages of apoptotic cells of the negative control, blank control and experimental groups were 5.88 +/- 0.99, 6.97 +/- 1.60, 16.40 +/- 1.95 respectively), along with an increased expression of activated caspase 3, and cell cycle inhibition at G(0)/G(1) phase (the relative number of cells at G(0)/G(1) phase were 43.65 +/- 3.44, 43.59 +/- 1.83 and 52.71 +/- 1.10, respectively). In addition, multinucleated giant cells were observed along with a marked inhibition of invasion as reflected by fewer penetrating cells by Boyden chamber assay (46.07 +/- 9.97, 47.87 +/- 9.58 and 38.67 +/- 6.59, respectively). CONCLUSIONS: Survivin expression is high in androgen-independent prostate cancer cells and likely may be related to the apoptosis, growth and invasion of the tumor cells. Targeting the survivin pathway by RNA interference appears to be a promising approach for clinical treatment of androgen-independent prostate cancer. PMID- 17134551 TI - [Changes of fibronectin and type IV collagen expression in cultured rat mesangial cells transfected with connective tissue growth factor expression vector]. AB - OBJECTIVE: To study the role of connective tissue growth factor (CTGF) in the development of glomerulosclerosis by experimental alteration of fibronectin (FN) and Type IV collagen (Col IV) expression in cultured rat mesangial cells (MsC). METHODS: CTGF expression vector was transfected into MsC by Lipofectimine method. Protein and mRNA expression levels of CTGF, FN and Col IV were studied by Western blot and reverse transcription-polymerase chain reaction (RT-PCR) respectively. RESULTS: Two of MsC clones (MCT-1 and MCT-2) with CTGF overexpression were successfully established and found to have significant increases of FN and Col IV at both protein and mRNA levels. Compared with the controls, the expression of FN protein and mRNA in the two clones were 3.2 times (P < 0.05) and 2.9 times (P < 0.05) higher respectively. The expression of Col IV protein and mRNA was 3.8 times (P < 0.01) and 2.4 times (P < 0.01) higher respectively. CONCLUSION: CTGF up-regulates FN and Col IV expression in MsC and may play an important role in the development of glomerulosclerosis. PMID- 17134552 TI - [Expression and significance of erythropoietin in human gastric carcinoma on tissue microarry]. PMID- 17134553 TI - [Regulating mechanism of stromal cell-derived factor-1 expression by hypoxia]. PMID- 17134554 TI - [Tumor suppressor gene VHL, hypoxia inducible factor, and renal cell carcinoma]. PMID- 17134555 TI - [Neuroendocrine differentiation in prostate cancer]. PMID- 17134556 TI - [Methodology of DNA staining by Azure A]. PMID- 17134557 TI - [Hyperplastic or neoplastic lesion of the spleen?]. PMID- 17134558 TI - [Mucinous tubular and spindle cell carcinoma of kidney]. PMID- 17134559 TI - [Multilocular cystic renal cell carcinoma]. PMID- 17134561 TI - [Epidermal growth factor receptor family and targeted anticancer therapy]. PMID- 17134562 TI - [Comparision of HER2/neu oncogene detected by chromogenic in-situ hybridization and immunohistochemistry in breast cancer]. AB - OBJECTIVE: Through comparison of HER2/neu oncogene detected by chromogenic in situ hybridization (CISH) and immunohistochemistry (IHC) in breast cancer, to explore the effect of CISH on detecting gene amplification of HER2. METHODS: Selected formalin-fixed paraffin-embedded breast samples whose pathological types were infiltrating ductal carcinomas (255 retrospective samples, 271 prospective samples), and these samples were detected by IHC and CISH. RESULTS: (1) In the retrospective study, CISH identified gene amplification in 91.6% of IHC score 3+ tumors (120/131) and in 56.5% of IHC score 2+ tumors (39/69), thus the concordant ratio between IHC and CISH was 81.2% (207/255). The two results showed significant correlation (P<0.01). (2) In the prospective study, the ratio of HER2 protein over expression detected by IHC was 31.7%, the ratio of HER2 gene amplification detected by CISH was 27.3%. CISH identified gene amplification in 91.4% of IHC score 3+ tumors (53/58) and in 46.4% of IHC score 2+ tumors (13/28), Concordant ratio between IHC and CISH was 89.7% (243/271). Two results showed significant correlation (P<0.01). (3) Paired CISH/FISH results were concordant in 14 of 15 cases. The remaining case was detected by FISH, but showed no HER2 gene amplification by CISH. (4) The gene amplification by CISH had a significantly reverse correlation with ER and PR expression (P<0.01). CONCLUSIONS: The results of HER2 gene amplification detected by CISH have high concordance with the results detectd by IHC and FISH. CISH is a novel technique for detecting HER2 gene amplification. PMID- 17134563 TI - [Analysis of HER2 gene status in breast cancer with HER2 protein overexpression]. AB - OBJECTIVE: To study the HER2 gene status (by fluorescence in situ hybridization (FISH) in breast cancer with HER2 protein overexpression, the correlation between gene amplification and protein overexpression, as well as the rate and significance of chromosome 17 aneusomy. METHODS: One hundred and twenty archival cases of breast cancer with formalin-fixed and paraffin-embedded tumor tissues with 2+ (42 cases) and 3+ (78 cases) HER2 protein overexpression by immunohistochemistry (IHC, HercepTest, Dako) were tested by FISH (PathVysion, Vysis) for HER2 gene status. The rate of chromosome 17 aneusomy was also analyzed. RESULTS: Amongst the 42 samples with IHC 2+, HER2 gene amplification was identified in 32 cases (76.19%), which included 11 cases with low amplification (ratio 2 approximately 4), 20 cases with moderate amplification (ratio 4 approximately 10) and 1 case with high amplification (ratio>10). Amongst the 78 samples with IHC 3+, HER2 gene amplification was identified in 71 cases (91.03%), which included 9 cases with low amplification, 48 cases with moderate amplification and 14 cases with high amplification. Chromosome 17 aneusomy was found in 83 cases (83/120, 69.17%), in which 14 cases (11.67%) showed hypodisomy (chromosome 17 copy number per cellor=3.76). Amongst the 42 cases with IHC 2+, 25 samples (59.52%) had chromosome 17 aneusomy, including 3 cases with hypodisomy, 18 cases with low polysomy and 4 cases with high polysomy. Amongst the 78 cases with IHC 3+, 58 samples (74.36%) had aneusomy 17, including 11 cases with hypodisomy, 34 cases with low polysomy and 13 cases with high polysomy. Most of IHC 2+ and FISH-positive cases had low or moderate HER2 gene amplification, while most of the IHC 3+ and FISH-positive cases had moderate or high gene amplification (P=0.0003). Six of the 7 samples with IHC 3+ and FISH-negativity had chromosome 17 aneusomy and 5 of the 10 samples with IHC 2+ and FISH-negativity had such aneusomy. CONCLUSIONS: A high concordance rate is noted between IHC 3+ and FISH positive results. The rate of FISH positive in IHC 2+ patients was higher than reported in other studies. Low or moderate HER2 gene amplification in IHC 2+ and moderate or high gene amplification in IHC 3+ occurs quite frequently. Chromosome 17 aneusomy (including hypodisomy, low polysomy and high polysomy) is also a relatively common phenomenon in our cohort with HER2 overexpression, with predominance of low polysomy. PMID- 17134565 TI - [Clinicopathologic study of solid papillary carcinoma of breast]. AB - OBJECTIVE: To study the clinicopathologic features and immunophenotype of solid papillary carcinoma (SPC) of breast. METHODS: Clinical and pathologic features of 21 cases of SPC, with or without stromal invasion, were analyzed. Immunohistochemical study (LSAB method, for cytokeratins, myoepithelial markers, chromogranin A, synaptophysin, Ki-67, estrogen receptor, progesterone receptor, c erbB-2 and pS2) and alcian blue staining were performed. RESULTS: All the patients were females with a mean age of 66.1 years. The clinical features were similar to those of classic papillary tumor. Metastasis was not observed in patients who had undergone axillary lymph node dissection. Histologically, the tumor displayed solid papillary growth pattern, with mucin production demonstrated in 19 cases. The tumor cells were oval, polygonal, spindled or signet ring-like and contained mildly to moderately pleomorphic nuclei. The mitotic count measured less than 5 per 10 high-power fields in 15 cases. Seven cases contained foci of invasive carcinoma which showed similar cytologic features as those of the in-situ component. Immunohistochemical study showed that the tumor cells expressed CK8 but not basal cell cytokeratin. Positivity for smooth muscle actin-alpha, calponin and p63 was demonstrated in the myoepithelial layers of fibrovascular cores, as well as around the expanded ductolobular units. Most cases also showed cytoplasmic positivity for chromogranin A (88.2%) and synaptophysin (82.4%). The proliferation index, as highlighted by Ki-67 immunostain, was 8.1%. The tumor expressed estrogen receptor, progesterone receptor and pS2. The staining for c-erbB-2 oncoprotein was negative. Follow up of 16 patients showed no evidence of recurrence or metastasis. CONCLUSIONS: SPC predominantly affects elderly females and has distinctive pathologic features and immunophenotype. Some cases of SPC are associated with mucinous and neuroendocrine components. Follow-up data suggest that SPC often carries an indolent clinical behavior and favorable prognosis. PMID- 17134566 TI - [Clinicopathologic study of breast cancer with features of endocrine ductal carcinoma-in-situ]. AB - OBJECTIVE: To study the clinicopathologic features and differential diagnosis of 18 cases of endocrine ductal carcinoma-in-situ (E-DCIS). METHODS: Eighteen cases of breast cancer with features of E-DCIS were studied by light microscopy, histochemistry and immunohistochemistry. E-DCIS was diagnosed if the histologic patterns were compatible with those described in the literature and at least 50% of the tumor cells expressing two of the three neuroendocrine markers employed (chromogranin, synaptophysin and neuron-specific enolase). RESULTS: E-DCIS tended to occur in older women. All the patients were over 61 years old (mean age=71 years). The presenting symptoms were either palpable breast mass or had nipple discharge. Histologically, E-DCIS demonstrated an expansile intraductal growth pattern. Intraductal papilloma was not uncommon at the peripheral area of the tumor. The tumor cells were polygonal, oval or spindle in shape and contained abundant eosinophilic to granular cytoplasm and mildly to moderately pleomorphic nuclei. Intracellular or extracellular mucin was highlighted by periodic acid Schiff (with diastase digestion) or alcian blue stains. Some tumor cells assumed a signet-ring configuration. All the three neuroendocrine markers were expressed by more than 50% of the E-DCIS cells. The neuroendocrine differentiation was further confirmed in some cases by CD57 and CD56 immunostaining. Pagetoid spread into adjacent ductolobular units was frequently seen in E-DCIS, and the expanded lobules were often not rimmed by myoepithelial cells. These two features helped to distinguish E-DCIS from usual ductal hyperplasia. CONCLUSIONS: E-DCIS represents a subgroup of low-grade DCIS, which carries characteristic morphologic features and immunophenotype. Conventional light microscopy usually permits a correct diagnosis. Ancillary histochemical and immunohistochemical studies can be helpful in doubtful cases. PMID- 17134567 TI - [Clinical and pathologic characteristics of primary angiosarcoma of breast]. AB - OBJECTIVE: To study the clinical and pathologic characteristics of primary angiosarcoma of breast. METHODS: Five cases of primary angiosarcoma of breast were reviewed. Immunohistochemical study (SP method) for factor VIII-related antigen, CD31, CD34, estrogen receptor, progesterone receptor and c-erbB-2 was performed. The clinical data were also analyzed. RESULTS: All of the 5 patients had a history of a painless breast lump. Ultrasound examination and mammogram did not reveal any specific findings. Histologically, various degree of cellular pleomorphism was noted. Immunohistochemical study showed a positive expression of endothelial associated antigens of the tumors cells in all the 5 cases. The staining result for estrogen receptor, progesterone receptor and c-erbB-2 was negative. CONCLUSIONS: Primary angiosarcoma of breast is a rare entity. The diagnosis is rather difficult owing to the lack of specific clinical findings and histological heterogeneity. Application of Positive expression of endothelial associated antigens of the tumors cells is helpful in making a pathological diagnosis morphologically. PMID- 17134568 TI - [Relationship between the malignant mesothelioma and simian virus 40 in China: a study of 17 cases]. AB - OBJECTIVE: To investigate whether simian virus 40 (SV40) was related to patients of malignant mesothelioma in China. METHODS: Paraffin-embeded samples of 17 patients with malignant mesothelioma were collected. After isolation of DNA from paraffin blocks, polymerase chain reaction (PCR) analyses were performed using three different sets of primer for detection of SV40 large T antigen gene. These samples were also immunohistochemically evaluated for expression of SV40 TAg protein with two different anti-SV40 Tag (Pab101 and Ab-2). RESULTS: Only one of the three primer pairs successfully amplified SV40 genome in three malignant mesothelioma samples. No immunopositive staining for SV40 TAg was found in any of the samples. CONCLUSIONS: The study shows that malignant mesothelioma in China may be independent of SV40 infection. PMID- 17134569 TI - [Clinicopathologic and immunophenotypic analysis of myeloid sarcoma]. AB - OBJECTIVE: To study the clinicopathologic features of myeloid sarcoma and to evaluate the role of immunohistochemical study in diagnosis of this entity. METHODS: Eighty-two cases of myeloid sarcoma were retrieved from the archives of Department of Pathology, West China Hospital of Sichuan University during the period from January, 1990 to February, 2005. The morphologic features were reviewed and classified according to the 2001 WHO classification for hematopoietic and lymphoid tissue tumors. Immunohistochemical study using a panel of 11 antibodies was performed on 73 cases. The survival data were collected and analyzed by SPSS 10.0. RESULTS: The median age of patients was 35.5 years. The male-to-female ratio was 1.4:1. The sites of occurrence included lymph node (43.1%), skin (16.7%), nose (7.8%), soft tissue (7.8%) and bone (6.9%). Fifty-one cases (62.2%) represented myeloid sarcoma associated with an underlying myeloproliferative disorder and 25 cases (30.5%) represented solitary myeloid sarcoma. As for the morphology, 79 cases (96.3%) were granulocytic sarcoma, including 41 cases (51.9%) blastic type, 25 cases (31.6%) immature type and 13 cases (16.5%) differentiated type. The other 3 cases (3.7%) were monoblastic sarcoma. Immature eosinophils were found in 51 cases (64.6%) of granulocytic sarcoma, among which 13 cases (31.7%) were of blastic type. Immunohistochemical study showed that 95.9% cases (70/73) were positive for myeloperoxidase, 95.5% (63/66) for lysozyme, 95.2% (60/63) for CD68 (KP1), 90.8% (59/65) for leukocyte common antigen, 85.7% (54/63) for CD43, 77.8% (49/63) for CD117, 58.7% (37/63) for CD99, 54.0% (34/63) for CD15, 22.2% (14/63) for CD34, and 4.7% (3/64) for CD68 (PG-M1). Proliferation index, as demonstrated by Ki-67 positivity, was 0.49+/-0.22. Follow-up data was obtained in 59 of the 82 patients. The two- and five-year survival rates were 36.1% and 17.3% respectively. No significant prognostic factors were found in the survival analysis. CONCLUSIONS: Myeloid sarcoma may precede, develop in a background of myeloproliferative disorder or even after remission of the disease. The presence of immature eosinophils is an important morphologic clue and immunohistochemical study plays an essential role in arriving at a correct diagnosis. Immunopositivity for myeloperoxidase is specific for granulocytic differentiation, while CD68 (PG-M1)-positivity suggests monocytic differentiation. Detailed clinicopathologic correlation is also helpful. PMID- 17134570 TI - [Follicular dendritic cell sarcoma: a clinicopathologic study of five cases]. AB - OBJECTIVE: To study the clinical pathological features and immunophenotype of follicular dendritic cell sarcoma (FDCS) with discussion on its diagnostic clues to improve diagnostic level. METHODS: Five cases of FDCS were analyzed by clinical, pathologic and immunohistochemistry methods. RESULTS: Five cases of FDCS were located in the cervical lymph node. Microscopically, the normal architectures were effaced by ovoid, spindle-shaped with fascicular, diffuse or whorled patterns and with rich lightly eosinophilic cytoplasm, syncytial appearance. Nuclei tend to show irregular clustering, scattered multinucleated giant cell. Nucleoli often distinct, sometimes prominent. Mitotic count variable, may show significant cellular pleomorphism. Immunohistochemical studies show that the tumor cells were positive for CD21, CD35, but negative for CD1a, CD34, CK and HMB45. Under electron microscopy, the tumor cells possessed long villus cytoplasmic processes and desmosome-like junctions, Birbeck granules were absent. CONCLUSIONS: FDCS is a rare malignant tumor and differential diagnosis includes Langerhans cell sarcoma, interdigitating dentric cell sarcoma, malignant fibrous histocytoma, melanoma, metastatic spindle cell carcinoma and others. Immunohistochemistry and electron microscopy are necessary for a correct diagnosis. PMID- 17134571 TI - [Expression of epithelial-cadherin, CD44v6 and connexin43 in hepatocellular carcinoma]. AB - OBJECTIVE: To study the expression of epithelial-cadherin (E-cad), CD44v6 and Cx43 in hepatocellular carcinoma (HCC) and its relationship with sex and age of patients, as well as tumor histopathologic grades. METHODS: Double immunofluorescent staining and laser scanning confocal microscopy was used to study the expression of E-cad, CD44v6 and Cx43 in 30 cases of normal liver tissue, 25 cases of benign hepatic lesions and 38 cases of HCC. In the HCC group, correlation of antigen expression with sex and age of patients and tumor histopathologic grades was studied by T-test. RESULTS: Significant decrease in expression of E-cad and Cx43 was noted in HCC group, as compared to normal liver tissue and benign hepatic lesion (P<0.05). On the other hand, CD44v6 expression was higher in HCC group than in the other two groups (P<0.05). In HCC group, the expression of E-cad and Cx43 did not correlate with sex, age and histopathologic grades (P>0.05). However, CD44v6 expression positively correlated with higher tumor histopathologic grades (P<0.05) but not sex and age of patients (P>0.05). In HCC group, the expression of E-cad positively correlated with that of Cx43, while the expression of CD44v6 negatively correlated with that of E-cad and Cx43. CONCLUSIONS: Tumor immunophenotype alters during development and progression of HCC. Low expression of E-cad and Cx43 and high expression of CD44v6 may be related to aggressive clinical behavior of HCC, moreover, high expression of CD44v6 correlated with high tumor grades. Detection of E-cad, CD44v6 and Cx43 expression may thus be useful in predicting prognosis of HCC. PMID- 17134572 TI - [Clinicopathologic study of ischemic intestinal disease due to mesenteric venous lesions]. AB - OBJECTIVE: To study the clinical and pathologic features of ischemic intestinal disease due to mesenteric phlebitis. METHOD: The clinical and pathologic features of the mesenteric venous lesions in 3 patients of ischemic intestinal disease admitted during the period from 2003 to 2004 were studied. RESULTS: All 3 patients had a clinical history of acute abdominal pain accompanying with a diffuse peritonitis. During operation, an infarcted intestinal segment was identified and was resected respectively in each patient. Histologic examination showed a lymphocytic infiltration and fibrinoid necrosis of the small to medium sized veins, associated with mural thrombosis and infarction of the corresponding intestinal wall and mesentery. The mesenteric arteries were spared. Two-year follow up of one case showed no evidence of local recurrence or systemic vasculitis. CONCLUSIONS: Ischemic intestinal disease due to mesenteric phlebitis is a rare entity with a pathological feature of inflammation of venous wall accompanying with the development of mural thrombosis and subsequent haemorrhagic infarction of intestine. The etiology is unknown and surgical resection of the involved intestinal segment is usually recommended. PMID- 17134573 TI - [Clinicopathologic study of aortic valves in children]. PMID- 17134574 TI - [Mutation of mitochondrial DNA in breast carcinoma]. PMID- 17134575 TI - [MicroRNA and cancer]. PMID- 17134576 TI - [Guideline of HER2 detection in breast cancer]. PMID- 17134577 TI - [Multiplex RT-PCR assay for detecting fusion genes of soft tissue small round cell tumors using paraffin-embedded and formalin-fixed tissue]. PMID- 17134578 TI - [Epithelioid angiosarcoma after radical mastectomy for breast carcinoma: a case report]. PMID- 17134579 TI - [Proximal epithelioid sarcoma: a case report]. PMID- 17134580 TI - [Post-transplant lymphoproliferative disorders: a report of two cases]. PMID- 17134581 TI - Clinical outcomes after autologous haematopoietic stem cell transplantation in patients with progressive multiple sclerosis. AB - BACKGROUND: Multiple sclerosis (MS) is a continuously disabling disease and it is unresponsive to high dose steroid and immunomodulation with disease progression. The autologous haematopoietic stem cell transplantation (ASCT) has been introduced in the treatment of refractory forms of multiple sclerosis. In this study, the clinical outcomes followed by ASCT were evaluated for patients with progressive MS. METHODS: Twenty-two patients with secondary progressive MS were treated with ASCT. Peripheral blood stem cells were obtained by leukapheresis after mobilization with granulocyte colony stimulating factor. Etoposide, melphalan, carmustin and cytosine arabinoside were administered as conditioning regimen. Outcomes were evaluated by the expanded disability status scale and progression free survival. No maintenance treatment was administered during a median follow-up of 39 months (range, 6 to 59 months). RESULTS: No death occurred following the treatment. The overall confirmed progression free survival rate was 77% up to 59 months after transplantation which was significantly higher compared with pre-transplantation (P = 0.000). Thirteen patients (59%) had remarkable improvement in neurological manifestations, four (18%) stabilized their disability status and five (23%) showed clinical recurrence of active symptoms. CONCLUSIONS: ASCT as a therapy is safe and available. It can improve or stabilize neurological manifestations in most patients with progressive MS following failure of conventional therapy. PMID- 17134582 TI - Enteral refeeding syndrome after long-term total parenteral nutrition. AB - BACKGROUND: Early enteral feeding (EF) may result in fever, elevated white blood cell count, increased serum levels of liver enzymes, and diarrhea. We name the complications "enteral refeeding syndrome", as a subtype of refeeding syndrome, because they are likely to result from long-term lack of lumen nutrition. The aim of this study was to investigate the characteristics of enteral refeeding syndrome after long-term total parenteral nutrition (TPN), and the solution for the disease. METHODS: We collected the clinical data of 100 patients with gastrointestinal fistula, who were cured from Apirl 2001 to July 2002. Their fasting time, daily stool frequency, body temperature, heart rate, respiratory rate, levels of transaminases, alkaline phosphatase (AKP), and gamma glutamylcyclotransferase (gamma-GT), white blood cell count, and systemic inflammatory reaction syndrome (SIRS) score were recorded before and 1, 3, 5, 10, and 15 days after EF. Student's t test and analysis of variance were used to analyze the data. RESULTS: Of the 100 patients, 56 were cured after selective resection of intestinal fistula, 15 were cured by emergency operation, and 29 recovered spontaneously. The levels of AKP and gamma-GT increased significantly on the 3rd day after EF [On the 3rd day after EF, (243.0 +/- 121.6) U/L and (177.2 +/- 109.9) U/L vs. before EF (181.5 +/- 127.5) U/L and (118.4 +/- 94.2) U/L, P < 0.05], and decreased gradually afterwards. The SIRS scores on the 1st day (1.05 +/- 1.08) and 3rd day (0.96 +/- 1.11) after EF were significantly higher than that before EF (0.72 +/- 0.84), then decreased to 0.83 +/- 0.91, 0.49 +/- 0.73 and 0.32 +/- 0.60 on the 5th, 10th and 15th days after EF. The number of patients with diarrhea at 1, 3, 5, 10 and 15 days post-EF were 31, 26, 12, 13, and 7, respectively. CONCLUSIONS: The longer the TPN lasts, the more severe the enteral refeeding syndrome becomes. Continuous EF is effective for the syndrome. Early enteral nutrition is useful in preventing it. PMID- 17134583 TI - Value of fructosamine measurement in pregnant women with abnormal glucose tolerance. AB - BACKGROUND: The concentration of serum fructosamine is correlated with plasma glucose level. The aim of this study was to determine whether the level of serum fructosamine can be diagnostic for abnormal glucose tolerance in pregnant women. METHODS: Serum samples were collected from 161 pregnant women between November 2004 and April 2005. The women were divided into three groups according to the gestational age (16 - 20 weeks group, 56 patients; 28 - 34 weeks group, 72; and 37 - 41 weeks group, 33). Each group was subdivided into normal and abnormal glucose tolerance subgroups. The levels of serum fructosamine were measured. Differences among the groups were assessed by ANOVA and Student-Newman-Keuls test. Correlations between the level of fructosamine and other variables including the results of glucose challenge test (GCT), oral glucose tolerance test (OGTT), and glycosylated hemoglobin (HbA1c) test, and infant's birth weight were analyzed by Pearson correlation. RESULTS: The level of serum fructosamine decreased with gestational age [(223.25 +/- 48.90) micromol/L, (98.44 +/- 29.57) micromol/L, and (53.99 +/- 29.94) micromol/L, respectively. P < 0.05]. It was higher in women with abnormal glucose tolerance than that in women with normal glucose tolerance, however, the difference reached statistical significance only in the 28 - 34 weeks group (P < 0.05). In this group, the level of serum fructosamine correlated positively with the GCT result (r = 0.28, P < 0.05). No correlation was found between fructosamine level and OGTT result, HbA1c level, or neonatal weight. CONCLUSIONS: Fructosamine can be used to monitor the glucose level of pregnant women with abnormal glucose tolerance, and to identify the patients at high risk of abnormal glucose tolerance, but can not be used to predict gestational diabetes mellitus (GDM) in early stage of pregnancy. PMID- 17134584 TI - Increased brachial-ankle pulse wave velocity is associated with impaired endothelial function in patients with coronary artery disease. AB - BACKGROUND: Pulse wave velocity and flow-mediated vasodilation (FMD) are widely used as noninvasive modalities for evaluating atherosclerosis. However, it is not known whether pulse wave velocity is related to FMD in patients with coronary artery disease (CAD). Therefore, the present study was designed to investigate the alteration in brachial-ankle pulse wave velocity (baPWV) and endothelial function in CAD patients. METHODS: Thirty-three patients with CAD and thirty control subjects were recruited for this study. baPWV was measured non-invasively using a VP 1000 automated PWV/ABI analyzer (PWV/ABI, Colin Co. Ltd., Komaki, Japan). Endothelial function as reflected by FMD in the brachial artery was assessed with a high-resolution ultrasound device. RESULTS: baPWV was increased in CAD patients compared with control subjects [(1756.1 +/- 253.1) cm/s vs (1495.3 +/- 202.3) cm/s, P < 0.01]. FMD was significantly reduced in CAD patients compared with control subjects [(5.2 +/- 2.1)% vs (11.1 +/- 4.4)%, P < 0.01]. baPWV correlated with FMD (r = -0.68, P < 0.001). The endothelium-independent vasodilation induced by sublingual nitroglycerin in the brachial artery was similar in the CAD group compared with the control group. CONCLUSIONS: CAD is associated with increased baPWV and endothelial dysfunction. Increased baPWV parallels diminished endothelial function. Our data therefore suggest that baPWV can be used as a noninvasive surrogate index in clinical evaluation of endothelial function. PMID- 17134585 TI - Effect of metabolic syndrome on prognosis and clinical characteristics of revascularization in patients with coronary artery disease. AB - BACKGROUND: People with metabolic syndrome are at higher risk for developing coronary artery disease (CAD). The effect of the metabolic syndrome on outcomes in patients with preexisting CAD has not been well studied. This study was conducted to assess the prevalence, characteristics, in hospital and long term prognosis of CAD with metabolic syndrome and to determine the factors influencing the prognosis of the disease. METHODS: The DESIRE registry contains data of 3696 patients with CAD between 2001 and 2004. Mean long term followup was (829 +/- 373) days. Diagnosis of metabolic syndrome was based on modified International Diabetes Federation (IDF) Worldwide Definition of the Metabolic Syndrome, using body mass index (BMI) instead of waist circumference. RESULTS: Of 2596 patients with complete records of height, weight, and so on, 1280 (49.3%) were identified with metabolic syndrome. The patients with metabolic syndrome had higher level of body mass index, systolic blood pressure, diastolic blood pressure, fasting glucose and disordered blood lipid (all P < 0.0001), with higher creatinine [(10.5 +/- 4.3) mg/L vs (9.9 +/- 2.9) mg/L, P < 0.0001] and the number of white blood cells [(7.49 +/- 2.86) x 10(9)/L vs (7.19 +/- 2.62) x 10(9)/L, P = 0.008) compared with those without metabolic syndrome. The patients with metabolic syndrome showed severer coronary angiographic alterations (left main artery and/or > or = 2-vessel) (73.6% vs 69.6%, P = 0.031). There were no significant differences of major adverse cardiac and cerebral events (MACCE) or mortality in hospital between the two groups. During followup, the ratio of MACCE in CAD with metabolic syndrome patients increased significantly (11.8% vs 10.0%, P = 0.044). Fasting blood glucose (> or = 1000 mg/L) and triglyceride (TG, > or = 1500 mg/L) were responsible for most of the increased risk associated with the metabolic syndrome (adjusted OR 1.465, 95% CI 1.037 - 1.874, P = 0.032; OR 1.378, 95% CI 1.014 - 1.768, P = 0.044). CONCLUSIONS: The prevalence of metabolic syndrome was very high in CAD patients. The metabolic syndrome confers a higher risk of long term MACCE in patients with CAD, and dysglycaemia and hypertriglycaemia appear to be responsible for most of the associated risk. PMID- 17134586 TI - Localization of cystathionine beta synthase in mice ovaries and its expression profile during follicular development. AB - BACKGROUND: In vitro fertilization (IVF) researches have suggested that cystathionine beta synthase (CBS) is involved in oocyte development. However, little is known about the regional and cellular expression patterns of CBS in the ovary. The purpose of this study was to analyze the localization of CBS in mice ovaries and to investigate the expression profile during follicular development. METHODS: We used in situ hybridization and immunohistochemical analysis to determine CBS expression in the ovaries of female Balb/c mice. Then the follicles were collected from F1 (C57BL x Balb/c) mice and cultured in vitro. With the method of semi-quantitative RT-PCR, we also investigated the expression profile of CBS during follicular development. RESULTS: CBS was absent in the oocytes, although it was ubiquitously expressed in the ovary with the strongest expression in follicular cells at all stages. In late antral follicles, CBS expression was markedly higher in granulosa cells located close to the antrum and in cumulus cells around the oocyte. The semi-quantitative RT-PCR showed that CBS mRNA was detected in follicles at all stages in vitro. In cumulus-oocyte complexes superovulated, CBS expression also increased rapidly. CONCLUSIONS: CBS was located mainly in the follicular cells in the ovaries. The level of CBS expression is high in follicles during folliculogenesis in mice. Differences in the CBS expression profile between oocyte and follicular cells suggest a role for CBS as a mediator in interactions between oocyte and granulosa cells. PMID- 17134587 TI - Screening of genes for proteins interacting with the PS1TP5 protein of hepatitis B virus: probing a human leukocyte cDNA library using the yeast two-hybrid system. AB - BACKGROUND: The hepatitis B virus (HBV) genome includes S, C, P and X regions. The S region is divided into four subregions of pre-pre-S, pre-S1, pre-S2 and S. PS1TP5 (human gene 5 transactivated by pre-S1 protein of HBV) is a novel target gene transactivated by the pre-S1 protein that has been screened with a suppression subtractive hybridization technique in our laboratory (GenBank accession: AY427953). In order to investigate the biological function of the PS1TP5 protein, we performed a yeast two-hybrid system 3 to screen proteins from a human leukocyte cDNA library interacting with the PS1TP5 protein. METHODS: The reverse transcription polymerase chain reaction (RT-PCR) was performed to amplify the gene of PS1TP5 from the mRNA of HepG2 cells and the gene was then cloned into the pGEM-T vector. After being sequenced and analyzed with Vector NTI 9.1 and NCBI BLAST software, the target gene of PS1TP5 was cut from the pGEM-T vector and cloned into a yeast expression plasmid pGBKT7, then "bait" plasmid pGBKT7-PS1TP5 was transformed into the yeast strain AH109. The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting hybridization. After expression of the pGBKT7-PS1TP5 fusion protein in the AH109 yeast strain was accomplished, a yeast two-hybrid screening was performed by mating AH109 with Y187 containing a leukocyte cDNA library plasmid. The mated yeast was plated on quadruple dropout medium and assayed for alpha-gal activity. The interaction between the PS1TP5 protein and the proteins obtained from positive colonies was further confirmed by repeating the yeast two-hybrid screen. After extracting and sequencing of plasmids from blue colonies we carried out a bioinformatic analysis. RESULTS: Forty true positive colonies were selected and sequenced, full length sequences were obtained and we searched for homologous DNA sequences from GenBank. Among the 40 positive colonies, 23 coding genes with known functions were obtained, including Homo sapien leukocyte adhesion protein p150, 95, interleukin 2 receptor gamma chain, PALM2-AKAP2 protein (PALM2-AKAP2), eukaryotic translation initiation factor 4A, beta-2-microglobin, solute carrier family 9 (sodium/hydrogen exchanger), calreticulin, asialoglycoprotein receptor 1 (ASGR1), MHC class II lymphocyte antigen, cytochrome c oxidase subunit 1, lymphocyte antigen 86 (LY86) and lymphocyte cytosolic protein 1. One novel gene with unknown function was found and named as PS1TP5BP1. After being electronically spliced, it was deposited in GenBank (accession number: DQ471327). CONCLUSIONS: Genes of proteins interacting with PS1TP5 were successfully screened from leukocyte cDNA library. These results suggested that PS1TP5 was closely correlated with immunoregulation, carbohydrate metabolism, signal transduction, the formation of hepatic fibrosis and initiation and development of tumors and also brought some new clues for further studying the biological functions of the pre-S1 protein. PMID- 17134588 TI - Effect of Astragalus membranaceus injection on the activity of the intestinal mucosal mast cells after hemorrhagic shock-reperfusion in rats. AB - BACKGROUND: The mechanism of mucosal damage induced by ischemia-reperfusion (IR) after hemorrhagic shock is complex; mast cells (MC) degranulation is associated with the mucosal damage. Astragalus membranaceus can protect intestinal mucosa against intestinal oxidative damage after hemorrhagic shock, and some antioxidant agents could prevent MC against degranulation. This study aimed to observe the effects of astragalus membranaceus injection on the activity of intestinal mucosal mast cells (IMMC) after hemorrhage shock-reperfusion in rats. METHODS: Thirty-two Wistar rats were randomly divided into the normal group, model group, low dosage group, (treated with Astragalus membranacaus injection, 10 g crude medication/kg) and high dosage group (treated with Astragalus membranacaus injection, 20 g crude medication/kg). The rat model of hemorrhagic shock reperfusion was induced by hemorrhage for 60 minutes followed by 90 minutes of reperfusion. The animals were administrated with 3 ml of the test drug solution before reperfusion. At the end of study, intestinal pathology, ultrastructure of IMMC, and expression of tryptase were assayed. The levels of malondisldehyde (MDA), TNF-alpha, histamine, and superoxide dismutase (SOD) activity in intestine were detected, and the number of IMMC was counted. RESULTS: The Chiu's score of the rats in the model group was higher than in other groups (P < 0.01). The Chiu's score in the high dosage group was higher than that in the low dosage group (P < 0.05). Hemorrhage-reperfusion induced IMMC degranulation: Astragalus membranaceus injection attenuated this degranulation. Expression of tryptase and the number of IMMC in the model group increased compared with the other groups (P < 0.01) and was significantly reduced by the treatments of Astragalus membranaceus injection at both doses. There was no significant difference between the two treatment groups (P > 0.05). MDA content and concentration of TNF-alpha in the model group were higher than that in the other three groups (P < 0.05), and the concentration of TNF-alpha in the low dosage group was higher than that in the high dosage group (P < 0.05). SOD activity and the concentration of histamine in the model group were lower than the other three groups (P < 0.05). There was a negative correlation between the Chiu's score and the concentration of histamine and a positive correlation between the Chiu's score and the concentration of TNF-alpha and between the SOD activity and the concentration of histamine in the four groups (P < 0.05). CONCLUSION: Astragalus membranaceus injection may reduce the damage to small intestine mucosa by inhibiting the activated IMMC after hemorrhagic shock. PMID- 17134589 TI - Type 1 iodothyronine deiodinase activity and mRNA expression in rat thyroid tissue with different iodine intakes. AB - BACKGROUND: Type 1 deiodinase (D1) plays an important role in the metabolism of thyroid hormone and has close relationship with thyroid function. In this study we explore the effects of iodine intake on D1 activity and its mRNA expression and its possible mechanism. METHODS: Forty-eight Wistar rats were randomly divided into six groups with 8 in each: low iodine (LI), normal iodine (NI), five fold iodine (HI(5)), ten-fold iodine (HI(10)), fifty-fold iodine (HI(50)), one hundred-fold iodine (HI(100)) group. Three months, six months and twelve months after admistration of potassium iodate, they were sacrificed and thyroids were excised. The expression of D1 mRNA in the thyroid tissue was determined by RT-PCR and D1 activity was analyzed by (125)I-rT3 as substrate. The thyroid hormone was measured with radioimmunoassay method. RESULTS: Compared with NI group, D1 mRNA expression in LI groups slightly decreased, and D1 activity greatly increased. Both T(3) and T(4) in thyroid tissue significantly decreased, but the T(3)/T(4) ratio increased. D1 mRNA expression decreased in all HI groups, and D1 activity was significantly lower in HI groups. There was a tendency of decrease in D1 activity with increased doses of iodine intakes. There was no significant difference in T(4) in thyroid tissue between HI groups and NI group, but a tendency of decrease in T(3) level was found in all HI groups. CONCLUSIONS: In the case of iodine deficiency, D1 activity increased greatly in order to convert more T(4) to T(3). Excess iodine can inhibit both D1 mRNA expression and its activity to protect organism from being injured by excessive T(3). PMID- 17134590 TI - Antimicrobial effect of acidified nitrate and nitrite on six common oral pathogens in vitro. AB - BACKGROUND: Salivary nitrate is positively correlated with plasma nitrate and its level is 9 times the plasma level after nitrate loading. Nitrate in saliva is known to be reduced to nitrite by oral bacteria. Nitrate and nitrite levels in saliva are 3 - 5 times those in serum in physiological conditions respectively in our previous study. The biological functions of high salivary nitrate and nitrite are still not well understood. The aim of this in vitro study was to investigate the antimicrobial effects of nitrate and nitrite on main oral pathogens under acidic conditions. METHODS: Six common oral pathogens including Streptococcus mutans NCTC 10449, Lactobacillus acidophilus ATCC 4646, Porphyromonas gingivalis ATCC 33277, Capnocytophaga gingivalis ATCC 33624, Fusobacterium nucleatum ATCC 10953, and Candida albicans ATCC 10231 were cultured in liquid medium. Sodium nitrate or sodium nitrite was added to the medium to final concentrations of 0, 0.5, 1, 2, and 10 mmol/L. All of the microorganisms were incubated for 24 to 48 hours. The optical densities (OD) of cell suspensions were determined and the cultures were transferred to solid nutrient broth medium to observe the minimum inhibitory concentration and minimum bactericidal/fungicidal concentration for the six tested pathogens. RESULTS: Nitrite at concentrations of 0.5 to 10 mmol/L had an inhibitory effect on all tested organisms at low pH values. The antimicrobial effect of nitrite increased with the acidity of the medium. Streptococcus mutans NCTC 10449 was highly sensitive to nitrite at low pH values. Lactobacillus acidophilus ATCC 4646 and Candida albicans ATCC 10231 were relatively resistant to acidified nitrite. Nitrate at the given concentrations and under acidic conditions had no inhibitory effect on the growth of any of the tested pathogens. CONCLUSION: Nitrite, at a concentration equal to that in human saliva, is both cytocidal and cytostatic to six principal oral pathogens in vitro, whereas nitrate at a similar concentration has no antimicrobial effect on these organisms. PMID- 17134591 TI - Lyophilized standards for the calibration of real time PCR assay for hepatitis C virus RNA. AB - BACKGROUND: Since October 1997, an international standard for hepatitis C virus (HCV) nucleic acid amplification technology assay, 96/790, has been available. We compared a series of lyophilized standards with known HCV RNA concentrations against the international standard in fluorescence quantitative PCR detection. METHODS: A series of lyophilized sera were calibrated by ROCHE COBAS AMPLICOR HCV Monitor test against the international standard and sent to various manufacturers to analyse the samples using their own kits. Then calibration curves from the series were compared with that obtained from the external standard calibration curve with the manufacture's series. RESULTS: The standard calibration curve with the series of lyophilized serum showed an excellent correlation (R(2) > 0.98), slope and intercept that were similar to those from the manufacture's series. When the standard calibration curve from the series of lyophilized standards were used to define the values of the given sample, lower coefficients of variation between kits from different manufactures were obtained. CONCLUSION: The results showed that the lyophilized standards could be used to setup the standard calibration curve for clinical HCV RNA quantitative PCR detection. PMID- 17134592 TI - Research on autosomal dominant polycystic kidney disease in China. AB - OBJECTIVE: To review the history and recent development of research on autosomal dominant polycystic kidney disease (ADPKD) in China. DATA SOURCES: Both Chinese and English literatures were searched in MEDLINE/CD ROM (1979 - 2006) and the Chinese Biomedical Literature Disk (1979 - 2006). STUDY SELECTION: Published articles about ADPKD from mainland of China were selected. Data were mainly extracted from 58 articles which are listed in the reference section of this review. RESULTS: Some preliminary reports on cyst decompression surgeries and mutation analysis represent the contribution to the ADPKD research from China in the history. A serial of basic research and clinical studies on ADPKD in recent years also have been summarized. A technique platform for ADPKD research was firstly established. The genomics/proteomics/bioinformatics approach was introduced, which provide a lot of valuable information for understanding the pathogenesis. By denature high performance liquid chromatography (DHPLC) technique the entire PKD1 and PKD2 gene sequence screening system for Chinese Han population has been successfully established. Based on the characteristic data of Chinese patients, an integrated therapy protocol was put forward and won an advantage over the traditional therapy. Some novel experimental studies on therapy also were encouraging. CONCLUSIONS: Remarkable progress of ADPKD research in China have been made recently. Still many works, including the government support, international collaboration and active participation of more Chinese nephrologists, should be enhanced to advance this process in the near future. PMID- 17134593 TI - Effect of ketogenic diet on hippocampus mossy fiber sprouting and GluR5 expression in kainic acid induced rat model. PMID- 17134594 TI - Polymorphism of the HLA-DQA1 and -DQB1 genes of Han population in Jiangsu Province, China. PMID- 17134595 TI - Renal cell carcinoma with vena caval tumor thrombus extending into the right atrium. PMID- 17134596 TI - Evaluation of the ventricular assist device programme in the UK. AB - OBJECTIVES: To summarise the relevant clinical effectiveness and cost effectiveness literature, to collect data on survival, transplantation rates, health-related quality of life (HRQoL) and resource use for ventricular assist device (VAD) and non-VAD transplant candidates in the UK, and to construct cost effectiveness and cost-utility models of VADs in a UK context. Also to investigate the factors that drive costs and survival. DESIGN: A comprehensive systematic review was carried out. Data were collected from April 2002 to December 2004, with follow-up to March 2005. Cost-effectiveness and cost-utility models of VAD devices were developed based on UK activity and outcomes collected from April 2002 to March 2005. SETTING: National Specialist Commissioning Advisory Group funded VAD implantation was carried out at the Freeman, Harefield and Papworth transplant centres in the UK. PARTICIPANTS: Seventy patients were implanted with a VAD as a bridge to transplantation between April 2002 and December 2004. Non-VAD-supported transplant candidates (n = 250), listed at the three centres between April 2002 and December 2004, were divided into an inotrope dependent group (n = 71) and a non-inotrope-dependent group (n = 179). Although patients in the inotrope-dependent group were closest to the VAD group they were less sick. The last group comprised a hypothetical worst case scenario, which assumed that all VAD patients would die in the intensive care unit (ICU) within 1 month without VAD technology. INTERVENTIONS: Patients were included who were implanted with a VAD designed for circulatory support for more than 30 days, with intention to bridge to transplantation. A multistate model of VAD and transplant activity was constructed; this was populated by data from the UK. MAIN OUTCOME MEASURES: Survival from VAD implant or from transplant listing for non-VAD patients to 31 March 2005. Serious adverse events and quality of life measures were used. Cognitive functioning was also assessed. Utility weights were derived from EuroQoL responses to estimate quality-adjusted life-years (QALYs). Incremental cost-effectiveness ratios (ICERs) were defined as the additional cost of VADs divided by additional QALYs. Time-horizons were 3 years, 10 years and the lifetime of the patients. RESULTS: Of 70 VAD patients, 30 (43%) died pretransplant, 31 (44%) underwent transplantation, and four (6%) recovered and had the VAD removed. Five patients (7%) were still supported for median of 279 days at the end of March 2005. Successful bridge-to-transplantation/recovery rates were consistent with published rates. Survival from VAD implantation was 74% at 30 days and 52% at 12 months. There were 320 non-fatal adverse events in 62 patients during 300 months of VAD support, mostly in the first month after implantation. Commonly observed events were bleeding, infection and respiratory dysfunction. Twenty-nine (41%) patients were discharged from hospital with a VAD. The 1-year survival post-transplantation was 84%. For the inotrope-dependent and non-inotrope-dependent transplant candidates, death rates while listed were 10% and 8% and the median waiting times were 16 and 87 days, respectively. For transplant recipients, 1-year survival was 85% and 84%, respectively. Both VAD and non-VAD patients demonstrated similar significant improvements in their New York Heart Association class after transplantation. All patients had poor EQ-5D pretransplantation; after transplantation the groups had similar EQ-5D of 0.76 irrespective of time after surgery. HRQoL was poor in the first month for VAD patients but better for those who waited longer in all groups. VAD patients reported more problems with sleep and rest and with ambulation in the first month. Symptom scores were similar in all groups pretransplant. After transplantation all groups showed a marked and similar improvement in physical and psychosocial function. Mean VAD implant cost, including device, was pound 63,830, with costs of VAD support for survivors of pound 21,696 in month 1 and pound 11,312 in month 2. Main cost drivers were device itself, staffing, ICU stay, hospital stay and events such as bleeding, stroke and infection. For the base case, extrapolating over the lifetime of the patients the mean cost for a VAD patient was pound 173,841, with mean survival of 5.63 years and mean QALYs of 3.27. Corresponding costs for inotrope-dependent patients were pound 130,905, with mean survival 8.62 years and mean QALYs 4.99. Since inotrope-dependent patients had lower costs and higher QALYs than VAD patients, this group is said to be dominant. Non-inotrope-dependent transplant candidates had similar survival rates to those on inotropes but lower costs, also dominant. Compared with the worst case scenario the mean lifetime ICER for VADs was pound 49,384 per QALY. In a range of sensitivity analyses this ranged from pound 35,121 if the device cost was zero to pound 49,384. Since neither inotrope-dependent transplant candidates nor the worst case scenario were considered fair controls the assumption was investigated that, without VAD technology, there would be a mixture of these situations. For mixtures considered the ICER for VADs ranged from pound 79,212 per QALY to the non-VAD group being both cheaper and more effective. CONCLUSIONS: There are insufficient data from either published studies or the current study to construct a fair comparison group for VADs. Overall survival of 52% is an excellent clinical achievement for those young patients with rapidly failing hearts. However, if the worst case scenario were plausible, and one could reliably extrapolate results to the lifetime of the patients, VADs would not be cost-effective at traditional thresholds. Further randomised controlled trials are required, using current second generation devices or subsequent devices and conducted in the UK. PMID- 17134597 TI - A systematic review and economic model of the clinical and cost-effectiveness of immunosuppressive therapy for renal transplantation in children. AB - OBJECTIVES: To review the clinical and cost-effectiveness of basiliximab, daclizumab, tacrolimus, mycophenolate mofetil (MMF), mycophenolate sodium (MPS) and sirolimus as possible immunosuppressive therapies for renal transplantation in children. DATA SOURCES: Electronic databases were searched up to November 2004. REVIEW METHODS: Data from selected studies were extracted and quality assessed. An economic model [Birmingham Sensitivity Analysis paediatrics (BSAp)] was produced based on an adaptation of a model previously developed for the assessment of the cost-effectiveness of immunosuppressants in adults following renal transplant. RESULTS: For the addition of basiliximab, one unpublished paediatric randomised control trial (RCT), reported that the addition of basiliximab to tacrolimus-based triple therapy (BTAS) failed to significantly improve 6-month biopsy-proven acute rejection (BPAR), graft function, graft loss and all-cause mortality. No significant difference between groups was seen in 6 month or 1-year or longer graft loss, all-cause mortality and side-effects. In a meta-analysis of adult RCTs, the addition of basiliximab to a ciclosporin, azathioprine and steroid regimen (CAS) significantly reduced short-term BPAR. There was no significant difference in short- or long-term graft loss, all-cause mortality or side-effects. One adult RCT was included for the addition of daclizumab to CAS, which reported reduced 1-year BPAR, although no difference between groups was seen in either 1- or 3-year graft loss, all-cause mortality and side-effects. For tacrolimus versus ciclosporin, one unpublished paediatric RCT found that a regimen of tacrolimus, azathioprine and a steroid (TAS) reduced 6-month BPAR and improved graft function [glomerular filtration rate (GFR)] compared with CAS. This improvement in BPAR with tacrolimus was as shown in the meta-analysis of adult RCTs. There was evidence, particularly in children, that in comparison with ciclosporin, tacrolimus may reduce long-term graft loss, although there is no benefit on total mortality. The total level of withdrawal in children was reduced in children receiving tacrolimus. Adult RCTs showed an increase in post-transplant diabetes mellitus with tacrolimus. For MMF versus azathioprine, a meta-analysis of adult RCTs showed MMF [regimen of ciclosporin, MMF and a steroid (CMS)] to reduce 1-year BPAR compared with azathioprine (CAS). There was evidence, particularly in children, that in comparison with azathioprine, tacrolimus may reduce long-term graft loss, although there is no benefit on total mortality. There was an increase in the level of cytomegalovirus infection with MMF, although the overall level of withdrawal due to adverse events was not different to that of azathioprine-treated adults. No study comparing MPS with azathioprine (CAS) was identified. In an adult RCT comparing MMF with MPS, there was no significant difference between groups in 1-year efficacy or side-effects. One unpublished paediatric RCT assessed the addition of sirolimus to CAS. BPAR, graft loss and all-cause mortality were not reported. In two adult RCTs, compared with azathioprine, sirolimus reduced 1-year BPAR, reduced graft function (as assessed by an increased serum creatinine) and increased the level of hyperlipidaemia. No significant differences were seen in other efficacy and side-effect outcomes. On an adult RCT comparing sirolimus with ciclosporin, there were no significant differences between groups in 1-year efficacy or side-effects with the exception of an increased level of hyperlipidaemia with sirolimus substitution. Both the assessment group and drug companies assessed the cost-effectiveness of the newer renal immunosuppressants currently licensed in children using an adaptation (BSAp) of the Birmingham Sensitivity Analysis (BSA) model. This model is based on a 10-year extrapolation of 1-year BPAR results sourced from paediatric RCTs or adult RCTs (where paediatric RCTs were not available). The addition of basiliximab and that of daclizumab to CAS was found to increase quality-adjusted life-years (QALYs) and decreased overall costs, a finding that was robust to sensitivity analyses. The incremental cost-effectiveness ratio (ICER) of replacing ciclosporin with tacrolimus was highly sensitive to the selection of the hazard ratio for graft loss from acute rejection, dialysis costs and the incorporation (or not) of side effects. The ICERs for tacrolimus versus ciclosporin ranged from about 46,000 pounds/QALY to about 146,000 pounds/QALY. Although sensitive to varying the hazard ratio for graft loss with acute rejection, the ICER for replacing azathioprine with MMF remained in excess of 55,000 pounds/QALY. CONCLUSIONS: In general, compared with a regimen of ciclosporin, azathioprine and steroid, the newer immunosuppressive agents consistently reduced the incidence of short-term biopsy-proven acute rejection. However, evidence of the impact on side-effects, long-term graft loss, compliance and overall health-related quality of life is limited. Cost-effectiveness was estimated based on the relationship between short term acute rejection levels from RCTs and long-term graft loss. Both the addition of daclizumab and that of basiliximab were found to be dominant strategies, that is, regarding cost savings and increased QALYs. The incremental cost effectiveness of tacrolimus relative to ciclosporin was highly sensitive to key model parameter values and therefore may well be a cost-effective strategy. The incremental cost-effectiveness of MMF compared with azathioprine, although also sensitive to model parameter, was unattractive. There is a particular need for RCTs to assess the use of MMF, MPS and daclizumab for renal transplantation in children where no such evidence currently exists. Future comparative studies need to report not only on the impact of the newer immunosuppressants on short- and long-term clinical outcomes but also on side-effects, compliance, healthcare resource, costs and health-related quality of life. PMID- 17134598 TI - Amniocentesis results: investigation of anxiety. The ARIA trial. AB - OBJECTIVES: The Amniocentesis Results: Investigation of Anxiety (ARIA) trial tested two hypotheses: first, that giving amniocentesis results out on a fixed date alters maternal anxiety during the waiting period, compared with a policy of telling parents that the result will be issued 'when available' (i.e. a variable date), and secondly, that issuing early results from a rapid molecular test alters maternal anxiety during the waiting period, compared with not receiving any results prior to the karyotype. The effects of the two interventions on anxiety 1 month after receiving karyotype results were also examined. DESIGN: A multi-centre, randomised, controlled, open fixed sample, 2 x 2 factorial design trial, with equal randomisation. SETTING: Twelve hospitals in England offering amniocentesis as a diagnostic test for Down's syndrome. PARTICIPANTS: A total of 226 women who had had an amniocentesis were randomised between June 2002 and July 2004. Eight women with abnormal results or test failure were excluded post randomisation. INTERVENTIONS: Issuing karyotype results on a prespecified fixed date, rather than issuing them as soon as they became available and issuing karyotype results alone, or subsequent to issuing results from a rapid molecular test for the most common chromosomal abnormalities. MAIN OUTCOME MEASURES: Average anxiety during the waiting period, calculated using daily scores from the short version of the Spielberger State-Trait Anxiety Inventory (STAI). Recalled anxiety, measured 1 month after receiving karyotype results, using a rating scale. Anxiety at the 1-month follow-up, measured using the short-form STAI. RESULTS: There was no evidence that giving out karyotype results on a fixed or on a variable date altered maternal anxiety during the waiting period. However, the analysis only had sufficient power to detect a moderate to large effect. Issuing early results from a partial, but rapid, test reduced maternal anxiety during the waiting period, compared with receiving only the full karyotype results. This was a moderate to large effect. In addition, group differences in recalled anxiety reflected fairly closely the differences in anxiety women had experienced while waiting for results. One month after receiving normal karyotype results, anxiety was low in all groups, but women who had been given rapid test results were more anxious than those who had not. This was a small to moderate effect. CONCLUSIONS: Since there are no clear advantages in anxiety terms of issuing karyotype results as soon as they become available, or on a fixed date, women could be given a choice between them. Rapid testing was a beneficial addition to karyotyping, at least in the short term. This does not necessarily imply that early results would be preferred to comprehensive ones if women had to choose between them. There should be further research, including more qualitative studies, into the causes, characteristics and consequences of anxiety associated with prenatal testing. The effects of different testing regimes on short- and long-term anxiety, on the preferences of women and on the relationship between anxiety and preference should be investigated. More research is needed on the ways in which information might be used to minimise anxiety in different testing regimes. Further research is also required into the policy implications of incorporating individual preferences for different testing regimes into prenatal testing programmes. PMID- 17134600 TI - [Can--and should--stress be treated?]. PMID- 17134601 TI - [Elective Cesarean section--a safe intervention?]. PMID- 17134602 TI - [Rituximab (MabThera) as treatment of active rheumatoid arthritis]. AB - Rituximab (RTX) is a murine/human monoclonal antibody to CD20, a protein expressed almost exclusively on human B-lymphocytes. RTX induces rapid and marked B-cell depletion with beneficial clinical effects in 1/3 to 1/2 of rheumatoid arthritis patients. Treatment is given as two iv. infusions with a two-week interval and in combination with methotrexate. Mild to moderate side-effects are frequent, particularly during the first infusion, but long-term side-effects are generally rare, although pulmonary events and reactivation of viral infections of the liver is of concern. PMID- 17134603 TI - [Gliomas in adults: primary non-surgical treatment]. AB - 900 to 1000 new cases of primary brain tumour occur each year in Denmark, and half of them are gliomas. The treatment is cytoreductive surgery, followed by radiation therapy and chemotherapy. Radiation therapy has a definite place in the treatment of low-grade gliomas, but the timing is a matter of dispute. Patients with Glioblastoma Multiform should be treated with concomitant radiation therapy and temozolamide, followed by adjuvant temozolamide. Anaplastic astrocytomas should be treated with postoperative radiation therapy with or without adjuvant chemotherapy. Anaplastic oligodendroglioma should be treated with radiation therapy only. PMID- 17134604 TI - [Elective Caesarean section team--an organizational innovation]. AB - INTRODUCTION: The increasing rate of delivery by caesarean section demands more effective use of resources in obstetrical departments and anaesthesiological departments. At the Danish State Hospital, Rigshospitalet, in 2002, we decided to optimise the cooperation between the various professionals involved, by softening the professional boundaries, and by performing most planned caesarean sections on the same day, carried out by the same team, the "elective caesarean section team" (EST). In 2003 a similar structure was established at Hvidovre Hospital. This paper describes the process of establishing EST, the organisational implications, and the results of an evaluation of EST by the users. MATERIALS AND METHODS: The case records of all caesarean sections performed by the EST-team at Rigshospitalet and Hvidovre Hospital in 2004, were examined. All users in a two month period evaluated the EST by questionnaires at planned discharge. RESULTS: Most users (60.5%) were discharged after two days. The user evaluation showed a high level of satisfaction. CONCLUSION: The introduction of EST has increased the efficiency within the organisation for carrying out planned caesarean sections. It has generated a high level of satisfaction among users, and a high educational value for the staff. PMID- 17134605 TI - [Complications of elective Caesarean section performed by a special section team]. AB - INTRODUCTION: Two Danish hospitals have introduced a new organizing structure according to which planned caesarean sections are performed by a team. Caesareans are normally carried out without complications, and this makes it possible to evaluate complications to the surgery isolatedly. The object of this article was to describe the medical complications in mother and child of planned caesarean section performed in team. MATERIALS AND METHODS: A total of 534 caesarean sections were performed in team in 2004. Information on complications to caesarean sections was obtained from medical records. RESULTS: The most frequent complication was blood loss of more than 1000 millilitres occurring in 2,2%. Other complications occurred with frequencies of about or less than 1%. 7% of the patients experienced a complication to caesarean performed in team. Respiratory morbidity occurred in 3.2% of the infants. CONCLUSION: Concerning the short-term consequences, there is a low rate of fetal and maternal complications in these women in the low-risk group. Long-term consequences are not expected. PMID- 17134606 TI - [Treatment of stress, sick leave and occupational stability]. AB - INTRODUCTION: The aim was to describe a rehabilitation process of individuals with stress-related adjustment disorders and analysing the prognostic impact of certain individual factors with sick leave and work resumption as outcome. METHODS: A retrospective cohort from the period 1996-2004 was conducted by means of an audit of 220 files of patients who have visited an occupational health care unit. RESULTS: A still increasing number of patients, mainly working in public health and welfare institutions, were admitted during the nine year study period. The average rehabilitation duration was 144 days (range: 3-689). After that period a decrease in the number of individuals on sick leave was found, and a total of 62% had returned to work or remained in work. The only individual predictor having negative impact on ability to work was psychopharmacologic drug treatment (Odds Ratio: 0.43: 0.23-0.81), whereas somatic drug treatment, age, gender, skill, workplace, matrimony, and smoking all were without any significant influence on work ability. CONCLUSION: This study shows that an occupational rehabilitation process of patients with stress-related adjustment disorders seems to improve ability to resume work, but a minor group, especially those in psychopharmacologic drug treatment, need prolonged rehabilitation. More rigorous study design is needed to confirm these findings. PMID- 17134607 TI - [Occupationally conditioned bladder cancer--are the patients identified?]. AB - INTRODUCTION: A minor percentage of bladder cancer cases in Denmark are caused by occupational exposure to carcinogens. The disease is therefore also present on the National Board of Industrial Injuries' list of occupational diseases, and reporting of these is compulsory for Danish physicians. In spite of this, only very few cases are reported annually. The aim of this study was to examine whether sufficiently detailed information on occupational exposure was obtained from the patient at the time of diagnosis, and if this presumed lack of information could be the reason for the possible low number of reporting. MATERIAL AND METHODS: We have examined 82 medical records from bladder cancer patients admitted or treated at Herlev University Hospital in the period from 01.01 2002 to 06.16 2004. General information, including information on occupation, was noted and examined. RESULTS: 48 charts lacked occupational history or had an insufficient degree of information so that a possible occupational disease could not be excluded. In the remaining 34 medical records, there were four patients with a positive exposure history which should, therefore, have been reported. There was no information in the medical records as to whether this had been done. CONCLUSION: Medical records of many bladder cancer patients lack sufficient information on occupational exposure. The four cases that should have been reported furthermore suggest that the physicians need to be educated on the known exposures and on how to report a patient with a positive exposure history. This could be a considerable reason for the few annually reported cases. PMID- 17134608 TI - [Stimulation with 0,3 mg recombinant human thyrotropin (rhTSH) increases the effect of 131I therapy in patients with nontoxic nodular goiter. A prospective, randomized, double-blind trial--secondary publication]. AB - In a randomized, double-blind, placebo-controlled trial, 57 patients with nontoxic nodular goiter were stimulated with either 0.3 mg recombinant human thyrotropin (rhTSH) or placebo before radioactive iodine 131I therapy. After one year the goiter reduction had improved by 35% compared to conventional 131I therapy. The difference was most pronounced in patients with a large goiter. Adverse effects, including development of permanent hypothyroidism, were significantly more frequent in the rhTSH group. Patient satisfaction was high and uninfluenced by the use of rhTSH. PMID- 17134609 TI - [Physical activity and clustering of CVD risk factors--secondary publication]. AB - This study aimed to derive guidelines on physical activity. We did a cross sectional study of 1732 9-year-old and 15-year-old children. Objectively measured physical activity was associated to clustered CVD risk with an OR for the least active of 3.29 compared with the most active quintile. The time spent in moderate activity in children where risk was elevated was less than 116 min per day in 9 year-old and less than 88 min per day in 15-year-old children. Physical activity levels should be higher than the current international guidelines of at least 1 h per day to prevent clustering of CVD risk factors. PMID- 17134610 TI - [Subacute intracoronary stent thrombosis in a patient with polycythaemia vera]. AB - Polycythaemia vera is a myeloproliferative syndrome with thromboembolic and haemorrhagic complications. We describe a patient with polycythaemia vera and acute coronary syndrome in whom two-vessel intracoronary stent thrombosis occurred a few hours after percutaneous coronary intervention. The case was successfully treated by repeated stenting and abciximab infusion. PMID- 17134611 TI - [Dynamic dermoscopy]. AB - Dermoscopy of early-stage melanoma can be challenging, and repeated examination at three-month intervals may disclose subtle changes. In patients with atypical nevus syndrome or more than 50 nevi, repetitive excision of benign lesions does not guarantee that melanomas will be identified at an early stage and exposes patients to potentially disfiguring surgery. We present the case of a high-risk patient where repeated dermoscopy of an in situ melanoma showed that part of the pigment network had coarsened, even though the lesion had not changed macroscopically. PMID- 17134613 TI - NIH awards nearly 4 million US dollars in support of bench-to-bedside research. PMID- 17134612 TI - [Picture of the month: influenza virus pneumonia]. PMID- 17134614 TI - Phase I and Phase II enzyme polymorphisms and childhood cancer. AB - Childhood cancers continue to be challenging clinical entities whose etiology, demographic characteristics, clinical progression, treatment efficacy, and outcomes remain incompletely understood. Research suggests that multiple environmental and genetic factors may play crucial roles in the pathophysiology of many of these malignancies. Recent attention has been directed to the role of carcinogen metabolizing enzymes in the etiology and progression of cancer in both adults and children due to their multitude of polymorphic variants and their intimate interaction with environmental factors. In particular, xenobiotic metabolizing enzymes (XME), which are intimately involved in the activation and deactivation of many environmental carcinogens, have become an area of significant interest. Traditionally, these enzymes have been classified into either phase I or phase II enzymes depending on their substrates, activity, and occasionally based on their sequence in the metabolic pathways, and have been demonstrated to have numerous polymorphic variants. Phase I enzymes predominantly consist of cytochrome enzymes responsible for mixed function oxidase activity, whereas phase II enzymes are frequently conjugation reactions necessary for drug metabolism or the further metabolism of phase I enzyme products. Current research has discovered numerous interactions between polymorphisms in these enzymes and changes in cancer susceptibility, treatment efficacy, and clinical outcomes in childhood cancer. Furthermore, studies of polymorphisms in these enzymes have demonstrated to have synergistic/antagonistic interactions with other XME polymorphisms and demonstrate variable influences on disease pathophysiology depending on the patient's ethnic background and environmental milieu. Continuing research on the role of polymorphisms in phase I and phase II enzymes will likely further elucidate the intimate role of these polymorphisms with environmental factors in the etiology of childhood cancer. PMID- 17134615 TI - Changes in central arterial pressure waveforms during the normal menstrual cycle. AB - BACKGROUND: Changes in estradiol and progesterone during the human menstrual cycle may impact vascular and cardiac function. Renin-angiotensin-aldosterone system (RAAS) hormones increase during the luteal phase of the menstrual cycle and may antagonize the vascular effects of estradiol. This study was designed to investigate central arterial changes, cardiac function, and RAAS activity in response to gonadal steroid variations during the menstrual cycle. METHODS: We studied 15 women during the follicular and midluteal phases with determination of estradiol, progesterone, hormones of the RAAS, and spot urine sodium and creatinine levels. Central pulsatile hemodynamics was evaluated using calibrated carotid tonometry and central aortic Doppler flow. Systolic ejection period (SEP) and systolic pressure time integral (SPTI) were computed from carotid pressure waveforms. RESULTS: Levels of estradiol, progesterone, and RAAS hormones were higher in the luteal phase. SEP and SPTI were lower during the luteal phase, whereas central and peripheral blood pressures and measures of arterial stiffness were unchanged between the two phases. The urine sodium-to-creatinine ratio was similar at both phases. CONCLUSION: Central arterial stiffness does not differ between the follicular and midluteal phases of the menstrual cycle in healthy women, despite significant changes in estradiol and progesterone levels. Systole was shortened during the midluteal phase. RAAS activation during the luteal phase may be responsible for a lack of the expected estradiol-mediated reduction in arterial stiffness between the two phases of the menstrual cycle. Because load was unchanged, the decrease in SEP and SPTI may represent a direct effect of estrogen, progesterone, or RAAS activation on ventricular function. PMID- 17134616 TI - Reengineering clinical research with informatics. AB - The future success of the translational research spectrum depends on the clinical research enterprise's ability to break through the barriers that constrain its productivity. As more basic science discoveries emerge, our ability to effectively translate this knowledge into improved patient care rests squarely on the manner in which we answer clinical questions. Informatics--the science of effective information use--is poised to help advance the conduct of science. However, incorporating informatics into the enterprise comes with its own set of challenges. To harness the benefits of improved information use, it is important to first establish how information flows within research. A thoughtful implementation of informatics--one that factors in social and organizational nuances--will undoubtedly lead to a more efficient and effective clinical research enterprise. PMID- 17134617 TI - Preparing data for analysis using microsoft Excel. AB - A critical component essential to good research is the accurate and efficient collection and preparation of data for analysis. Most medical researchers have little or no training in data management, often causing not only excessive time spent cleaning data but also a risk that the data set contains collection or recording errors. The implementation of simple guidelines based on techniques used by professional data management teams will save researchers time and money and result in a data set better suited to answer research questions. Because Microsoft Excel is often used by researchers to collect data, specific techniques that can be implemented in Excel are presented. PMID- 17134618 TI - Prognostic importance of wall motion abnormalities in patients with ischemic cardiomyopathy and an implantable cardioverter-defibrillator. AB - Patients with extensive regional wall motion abnormalities are predisposed to development of ventricular tachyarrhythmia. The prognostic effect of this in patients with an implantable cardioverter-defibrillator (ICD) and coronary artery disease (CAD) is not known. Echocardiographic left ventricular systolic indexes, wall motion score index (WMSI), and extent of regional akinesia in 140 patients (65 +/- 10 years old; 92% men) with an ICD and CAD were studied. Arrhythmic events requiring ICD therapy and causing death (n = 41, 29%) were recorded over a mean follow-up of 1.4 +/- 0.8 years. Left ventricular basal fractional shortening, ejection fraction, global WMSI, and extent of akinesia, especially in the inferoposterior regions of a right coronary artery territory, were univariate predictors (all p values <0.05). Global WMSI (hazard ratio 2.18, 95% confidence interval 1.03 to 4.65, p = 0.04) and fractional shortening (hazard ratio 0.93, 95% confidence interval 0.88 to 1.00, p = 0.04) were multivariate predictors. Global WMSI (p = 0.04) and > or =2 right coronary region akinetic segments (p = 0.05) provided incremental risk prediction to left ventricular ejection fraction in a global risk-assessment model (chi-square p = 0.001). Presence of right coronary region akinesia better identified those at increased risk of events (p = 0.02) compared with the presence of left anterior descending region akinesia (p = 0.2), independent of systolic function. In conclusion, global WMSI and left ventricular basal fractional shortening were important additional risk predictors of ICD events in CAD. Global WMSI and right coronary region inferoposterior akinesia provided independent and incremental risk assessment to left ventricular ejection fraction and improved identification of those at increased risk of ICD related events in patients with ischemic cardiomyopathy. PMID- 17134619 TI - Predictors of coronary artery disease in patients with left bundle branch block undergoing coronary angiography. AB - Patients with left bundle branch block (LBBB) and concomitant coronary artery disease (CAD) have a worse prognosis than those with LBBB without CAD. In addition, subjects with CAD and concomitant LBBB have a higher cardiovascular mortality than those with a similar extent of CAD but without LBBB. Because the presence of LBBB makes the noninvasive identification of CAD problematic, patients with LBBB often are referred for coronary angiography to assess the presence and severity of CAD. To determine the clinical and demographic variables that might help identify those with CAD, we analyzed data from 336 consecutive patients with LBBB referred for coronary angiography. Of the 336, 54% had CAD. In conclusion, those with CAD were likely to be older, Caucasian, and men; they were more likely to have angina pectoris, myocardial infarction, and diabetes mellitus; and they were more likely to have a left ventricular ejection fraction <0.50. In contrast, patients with heart failure were less likely to have CAD. PMID- 17134620 TI - Usefulness of nocturia as a mortality risk factor for coronary heart disease among persons born in 1920 or 1921. AB - Cardiovascular events are clustered in the morning hours, after increases in blood pressure and heart rate that accompany awakening and arising. Similar hemodynamic changes occur during the night after nocturnal awakening and getting up. Such changes are common among older patients who have nocturia frequently and rise to urinate. We tested the hypothesis that nocturia may be associated with increased mortality in a population sample of 456 subjects born from 1920 to 1921, examined in 1990, and followed for total mortality until 2002. At baseline, they were questioned about nocturia (> or =2 times at night) as part of a detailed questionnaire and examination. Twelve-year survival was significantly lower (61% vs 72%, p = 0.0206) among subjects reporting nocturia (n = 160, 64% men) compared with those without nocturia (n = 296, 50% men). After accounting for numerous confounders, a proportional hazard model determined the mortality hazard ratio (HR) for nocturia alone to be 0.89 (95% confidence interval [CI] 0.55 to 1.43). The interaction between nocturia and previous coronary heart disease (CHD) was highly significant (p <0.0001), with an interaction variable HR of 2.16 (95% CI 1.01 to 4.61). Survival of patients who had CHD with nocturia (n = 54) versus those without nocturia (n = 65) was 44% versus 66% (p = 0.0201). Among patients with CHD, the mortality HR for nocturia was 2.11 (95% CI 1.16 to 4.00). In conclusion, nocturia is a significant independent predictor of mortality among 70-year-old patients with known CHD and thus warrants special attention. PMID- 17134621 TI - Usefulness of vasopressin administered with epinephrine during out-of-hospital cardiac arrest. AB - Vasopressin administration has been suggested during cardiopulmonary resuscitation, and a previous clinical trial has suggested that vasopressin is most effective when administered with epinephrine. Adult subjects (n = 325) who received > or =1 dose of intravenous epinephrine during cardiopulmonary resuscitation for nontraumatic, out-of-hospital cardiac arrest were randomly assigned to receive 40 IU of vasopressin (n = 167) or placebo (n = 158) as soon as possible after the first dose of epinephrine. The rate of return of pulses was similar between the vasopressin and placebo groups (31% vs 30%), as was the presence of pulses at the emergency department (19% vs 23%). No subgroup appeared to be differentially affected, and no effect of vasopressin was evident after adjustment for other clinical variables. Additional open-label vasopressin was administered by a physician after the study drug for 19 subjects in the placebo group and 27 subjects in the vasopressin group. Results were similar if these subjects were excluded or were assigned to an actual drug received. Survival duration for subjects admitted to the hospital did not differ between groups. In conclusion, vasopressin administered with epinephrine does not increase the rate of return of spontaneous circulation. PMID- 17134622 TI - Brain natriuretic peptide and other risk markers for outcome assessment in patients with non-ST-elevation coronary syndromes and preserved systolic function. AB - Several emerging cardiac markers constitute strong predictors among patients with coronary artery disease. In particular, brain natriuretic peptide (BNP), troponin T (TnT), and C-reactive protein (CRP) are related to increased risk of recurrent ischemic events and death. However, little is known about the utility of these biomarkers in combination. This study examined risk assessment in patients with coronary artery disease and preserved systolic function. We studied 208 consecutive patients (138 men, 70 women) with stable angina, unstable angina, and non-Q-wave myocardial infarction whose plasma BNP, TnT, and CRP levels were measured at hospital admission. All recruited patients underwent echocardiographic examination, and selective coronary angiography was performed. After adjusting for clinical presentation, age, gender, and common risk factors, BNP was demonstrated as a strong predictor of heart failure (6 months, odds ratio [OR] 2.03, 95% confidence interval [CI] 1.24 to 2.9, p <0.01; 12 months, OR 2.65, 95% CI 1.69 to 3.5, p <0.001) and mortality at 3, 6, and 12 months (p <0.001). BNP was also significantly related to extent of coronary artery disease and left anterior descending artery involvement (p <0.01). Patients with a BNP level >80 pg/ml in all 3 groups had a significantly poorer prognosis with increased incidence of heart failure and death. CRP was related to recurrent ischemic events (infarct or recurrent angina, OR 1.4, 95% CI 1.14 to 2.08, p <0.01) and was associated with major cardiac revascularization at 12 months (OR 1.51, 95% CI 1.29 to 1.73, p <0.001). TnT demonstrated a mild correlation with recurrent infarct or angina at 12 months (OR 1.1, 95% CI 0.96 to 1.22, p <0.05) but appeared related to multivessel coronary artery disease (OR 1.47, 95% CI 1.05 to 1.99, p <0.01). In conclusion, BNP appears to be associated with a long-term increased risk of mortality and heart failure in patients with apparently mild risk. BNP is also associated with a larger extent and greater severity of myocardial ischemia. Early BNP measurement could provide incremental information to TnT and CRP, and it may be the strongest independent predictor of cardiac outcome in subjects without left ventricular dysfunction or enlargement. PMID- 17134623 TI - Comparison of early mortality of paramedic-diagnosed ST-segment elevation myocardial infarction with immediate transport to a designated primary percutaneous coronary intervention center to that of similar patients transported to the nearest hospital. AB - Speed of reperfusion is critical in ST-segment elevation myocardial infarction (STEMI). We assessed the safety and feasibility of an integrated metropolitan approach in which advanced-care paramedics interpret the prehospital electrocardiogram and independently refer patients with STEMI to a designated center for primary percutaneous coronary intervention (PCI). We developed and implemented a protocol in which paramedics trained in electrocardiographic interpretation bypassed the nearest emergency room and referred patients with suspected STEMI directly to a designated primary PCI center (paramedic-referred primary PCI). Outcomes of these patients were compared with those of a retrospective cohort of 225 consecutive patients with STEMI transported by ambulance to the nearest hospital emergency department. We treated 108 consecutive patients with STEMI using ambulance services according to the paramedic-referred primary PCI protocol. Primary PCI was performed in 93.5% versus 8.9% in the control group, and the median door-to-balloon time was 63 versus 125 minutes in the control group (p <0.0001 for 2 comparisons). Thrombolytic therapy was prescribed to 80.4% of the control group, with a median door-to-needle time of 41 minutes. In-hospital mortality was 1.9% in the paramedic-referred primary PCI group versus 8.9% in the control group (p = 0.017) and remained significantly lower after statistical adjustment for baseline risk. In conclusion, paramedic-referred primary PCI is a safe and feasible strategy for treating STEMI that is associated with rapid and effective reperfusion and very low in-hospital mortality. PMID- 17134624 TI - Comparison of outcomes of percutaneous coronary interventions in patients of three age groups (<60, 60 to 80, and >80 years) (from the New York State Angioplasty Registry). AB - Octogenarians have been under-represented in percutaneous coronary intervention (PCI) trials despite an increase in referrals for PCI. As the United States population ages, the number of high-risk PCIs in the elderly will continue to increase. This study investigated the effect of age on short-term prognosis after PCI in 3 age groups. Using the 2000/2001 New York State Angioplasty Registry, we compared in-hospital mortality and major adverse cardiac events (MACEs; death, stroke, or coronary artery bypass grafting) in emergency and elective PCI cohorts across 3 age categories of patients: 10,964 patients who underwent emergency PCI (<60 years of age, n = 5,354; 60 to 80 years of age, n = 4,939; >80 years of age, n = 671) and 71,176 patients who underwent elective PCI (<60 years of age, n = 24,525; 60 to 80 years of age, n = 40,869; >80 years of age, n = 5,782). Patients were considered to have undergone an emergency PCI if they had an acute myocardial infarction within 24 hours, had thrombolytic therapy within 7 days, or presented with hemodynamic instability or shock. Elderly patients had more co morbidities, including more extensive coronary atherosclerosis, hypertension, peripheral vascular disease, and renal insufficiency, and presented more frequently with hemodynamic instability or shock. In the emergency PCI group, in hospital mortality (1.0% vs 4.1% vs 11.5%, p <0.05) and MACEs (1.6% vs 5.2% vs 13.1%, p <0.05) increased incrementally by age group. In the elective PCI group, rates of in-hospital complications were considerably lower, with an incremental increase in mortality (0.1% vs 0.4% vs 1.1%, p <0.05) and MACEs (0.4% vs 0.7% vs 1.6%, p <0.05). Age was strongly predictive of in-hospital mortality for emergency and elective PCI by multivariate analysis. In conclusion, elective PCI in the elderly has favorable outcome and acceptable short-term mortality in the stent era. Elderly patients, in particular octogenarians undergoing emergency PCI, have a substantially higher risk of in-hospital death. PMID- 17134625 TI - Intravascular brachytherapy versus drug-eluting stents for the treatment of patients with drug-eluting stent restenosis. AB - Drug-eluting stents (DESs), although promising technology, still are associated with restenosis; therefore, we evaluated the safety and efficacy of intravascular radiation therapy for the treatment of DES in-stent restenosis (ISR). Treatment of DES ISR has not been established, although intravascular radiation therapy is an effective treatment for patients with ISR of bare metal stents. Other modalities are conventional percutaneous coronary intervention (PCI), including restenting with DES. Radiation for Eluting Stents in Coronary FailUrE (RESCUE) is an international, Internet-based registry of 61 patients who presented with ISR of a DES and were assigned to intravascular radiation therapy with commercially available systems after PCI. Outcomes of these patients were compared with those of a consecutive series of 50 patients who presented with ISR of a DES and were assigned to repeat DES (r-DES) treatment. Baseline clinical and angiographic characteristics were similar between groups, except for more Cypher stents as the initial DES that restenosed in the r-DES group than in the intravascular radiation therapy group (88.5% vs 69%, p = 0.01). At 8 months there were fewer overall major adverse cardiac events in the intravascular radiation therapy group compared with the r-DES group (9.8% vs 24%, p = 0.044). The need for target vessel and target lesion revascularizations was similar in the 2 groups at 8 months. There has been no report of subacute thrombosis in either group. In conclusion, intravascular radiation therapy as adjunct therapy to PCI for patients presenting with ISR of a DES is safe and should be considered an alternative therapeutic option for this difficult subset of patients. PMID- 17134626 TI - Early- and medium-term outcomes after paclitaxel-eluting stent implantation for sirolimus-eluting stent failure. AB - The optimal treatment for sirolimus-eluting stent (SES) restenosis is not known. This study evaluated the safety and clinical outcome of paclitaxel-eluting stent (PES) implantation for SES restenosis. From March 2004 to July 2005, PESs were implanted in 125 patients with 140 lesions with SES restenosis. Acute and 6-month clinical outcomes were determined through review of the medical record and/or telephone interview. In-hospital major adverse cardiac events (death, nonfatal myocardial infarction, or repeat revascularization) occurred in 14 patients (11.2%), driven entirely by postprocedure non-Q-wave myocardial infarction. At a mean clinical follow-up of 7.2 +/- 1.8 months, the incidence of target lesion revascularization (TLR) was 14.0%, and the rate of major adverse cardiac events was 17.2%. Subacute thrombosis occurred in 2 patients (1.6%). Length of PES implanted, postprocedure diameter stenosis, and total occlusion of the target lesion were independent predictors of TLR. In patients with de novo SES restenosis, TLR was only 8.7%. In conclusion, at medium-term follow-up, PES implantation for SES failure appears to be safe and effective, although efficacy is decreased in the setting of total occlusions, greater residual diameter stenosis, and longer PESs. PMID- 17134627 TI - Effects of glucose-insulin-potassium infusion on myocardial perfusion and left ventricular remodeling in patients treated with primary angioplasty for ST elevation acute myocardial infarction. AB - The role of glucose-insulin-potassium (GIK) infusion in the management of acute coronary syndrome is controversial. Limited data are available on the effects of adjunctive high-dose GIK (30% glucose, 50 IU of insulin, 80 mEq of potassium chloride infused at 1.5 ml/kg/hour over 24 hours) on myocardial perfusion and left ventricular (LV) remodeling in patients treated with primary percutaneous coronary intervention (PCI) for ST-segment elevation myocardial infarction. In this prospective study, 73 patients were randomized to receive GIK infusion (n = 40) or saline (placebo, n = 33) in addition to standard therapy. The primary end points were myocardial perfusion after PCI and LV remodeling at 6 months. Thrombolysis In Myocardial Infarction frame count and myocardial blush grade were evaluated before and after reperfusion treatment. LV end-diastolic and end systolic volumes, ejection fraction, and wall motion score index were assessed in each patient after PCI and after 6 months. Although no differences in final Thrombolysis In Myocardial Infarction flow were observed between the 2 groups, myocardial blush grade 3 was more frequently achieved in the GIK group (p <0.05). At 6 months, ventricular remodeling was more often observed in the control group (24% vs 3%, p <0.05). In conclusion, GIK infusion in adjunct to primary PCI in patients with ST-segment elevation myocardial infarction was safe, improved myocardial perfusion after revascularization, and was associated with less LV remodeling at follow-up. PMID- 17134628 TI - Comparison of the effects of nicorandil versus verapamil on the incidence of slow flow/no reflow during rotational atherectomy. AB - In rotational atherectomy, the slow-flow/no-reflow phenomenon is a common complication, which usually results in myocardial injury. We prospectively randomized 111 patients with successful rotational atherectomy into a verapamil group (group V, n = 56) and a nicorandil group (group N, n = 55). We compared final burr size, total ablation time, maximum decrease in revolutions per minute, incidence of slow flow/no reflow, creatine kinase-MB increase, and cardiac troponin T increase between the 2 groups. Patient and procedural characteristics were not significantly different between groups. The incidence of no reflow/slow flow was significantly lower in group N than in group V (3.6% vs 17.9%, respectively, p <0.05). The incidence of a minor increase in cardiac markers was also significantly lower in group N than in group V (creatine kinase-MB 3.6% vs 16.1%, p <0.05; cardiac troponin T 5.5% vs 21.4%, p <0.05). In conclusion, our results showed that nicorandil decreases the incidence of slow flow/no reflow during rotational atherectomy. PMID- 17134629 TI - Value of prolonged QRS duration as a predictor of low cardiac output syndrome in patients with impaired left ventricular systolic function who undergo isolated coronary artery bypass grafting. AB - This study determined whether prolonged QRS duration (QRSd; > or =120 ms) is an independent predictor of low cardiac output syndrome (LCOS) in patients with low left ventricular (LV) ejection fraction (EF) who underwent isolated coronary artery bypass grafting (CABG). Abnormal LV systolic function places patients at greater risk for developing LCOS after isolated CABG. In patients with this form of ventricular function impairment, prolonged QRSd is associated with adverse hemodynamic effects. Clinical, operative, and outcome data from 190 consecutive patients with LVEF <50% who underwent isolated CABG (mean 62 +/- 9 years of age) were retrospectively analyzed. For all patients, preoperative QRSd was determined. LCOS was the primary outcome investigated. Fifty-seven patients (30%) developed LCOS. Compared with the subgroup without LCOS, the subgroup with this syndrome had significantly larger proportions of patients with LVEF <30% and prolonged QRSd. In addition, the group that developed LCOS had a longer mean QRSd (117 +/- 25 vs 102 +/- 17 ms, respectively, p = 0.00003) and a significantly higher frequency of adverse postoperative outcomes. Hospital stay was significantly longer in the subgroup with LCOS than in the subgroup without. Multivariate logistic regression analysis identified prolonged QRSd as the most significant predictor of LCOS. LVEF <30%, diuretic therapy, and preoperative risk score (European System for Cardiac Operative Risk Evaluation) were also identified as independent predictors of LCOS. In conclusion, in patients with impaired LV systolic function, prolonged QRSd is a highly significant predictor of LCOS development after isolated CABG. PMID- 17134630 TI - Non-high-density lipoprotein and very-low-density lipoprotein cholesterol and their risk predictive values in coronary heart disease. AB - To determine if non-high-density lipoprotein (HDL) cholesterol is a more useful predictor of coronary heart disease (CHD) risk than low-density lipoprotein (LDL) cholesterol and if very-low-density lipoprotein (VLDL) cholesterol is an independent predictor of CHD risk, data from the Framingham Heart Study (2,693 men, 3,101 women) were used for this analysis. All subjects were aged > or =30 years and free of CHD at baseline, and incident CHD was the end point (618 men, 372 women). Cox proportional-hazards models were used to assess the risk for CHD (relative risks and 95% confidence intervals) on the basis of the joint distribution of LDL cholesterol and non-HDL cholesterol (in milligrams per deciliter), as well as LDL cholesterol, non-HDL cholesterol, and VLDL cholesterol as continuous variables. After multivariate adjustment, within non-HDL cholesterol level, no association was found between LDL cholesterol and the risk for CHD, whereas within LDL cholesterol levels, a strong positive and graded association between non-HDL cholesterol and risk for CHD was observed. When the analysis was repeated within triglyceride levels (<200 vs > or =200 mg/dl), the risk pattern did not change significantly. Also, VLDL cholesterol was found to be a significant predictor of CHD risk after adjusting for LDL cholesterol at triglyceride levels of > or =200 or <200 mg/dl. In conclusion, these results suggest that non-HDL cholesterol level is a stronger predictor of CHD risk than LDL cholesterol; that is, VLDL cholesterol may play a critical role in the development of CHD. PMID- 17134631 TI - Comparison of apolipoprotein-B/apolipoprotein-AI in subjects with versus without the metabolic syndrome. AB - Recent studies have suggested that the apolipoprotein-B (apo-B)/apolipoprotein-AI (apo-AI) ratio predicts cardiovascular risk better than any of the cholesterol indexes. The aim of the present study was to assess if the apo-B/apo-AI ratio is related to the metabolic syndrome and its components. Data were analyzed from 2,964 subjects (mean age 48 years; 1,516 men, 1,448 women) from the National Health and Nutrition Examination Survey III with apolipoprotein data who were evaluated for the metabolic syndrome and its components. The metabolic syndrome was defined according to the criteria of the National Cholesterol Education Program Adult Treatment Panel III and the International Diabetes Federation. The mean values of the apo-B/apo-AI ratio in subjects with and without the metabolic syndrome were compared. Overall, the median distribution of the apo-B/apo-AI ratio was significantly greater (p <0.0001) in subjects with the Adult Treatment Panel III metabolic syndrome (0.90) than without (0.69). The apo-B/apo-AI ratio was associated significantly with each of the metabolic syndrome components, in descending order of magnitude: low high-density lipoprotein cholesterol (odds ratio [OR] 5.7), high triglycerides (OR 4.7), high waist circumference (OR 2.6), high fasting glucose (OR 1.9), and high blood pressure (OR 1.5). The apo-B/apo-AI ratio was also different between subjects with and without the metabolic syndrome. Mean values of apo-B/apo-AI increased significantly as the numbers of metabolic syndrome components increased in men (p <0.0001) and women (p <0.0001). After excluding high-density lipoprotein cholesterol and triglycerides as criteria for the metabolic syndrome, the association between means persisted (analysis of variance p <0.0001) in men and women. Apo-B/apo-AI was significantly associated with the presence of the metabolic syndrome (OR 5.1, p <0.0001). In conclusion, the apo-B/apo-AI ratio is strongly associated with the presence of individual metabolic syndrome components, with the metabolic syndrome itself, and with insulin resistance. An elevated apo-B/apo-AI ratio may constitute an important feature of the metabolic syndrome and may provide an additional mechanism to explain the increased cardiovascular risk in subjects with this syndrome. PMID- 17134632 TI - Ultrasound assessment of subclinical cardiovascular disease in a community-based multiethnic population and comparison to the Framingham score. AB - The presence of subclinical cardiovascular disease has been documented to indicate high coronary risk. This study investigated the impact of subclinical cardiovascular disease derived from echocardiography and carotid ultrasonography on traditional coronary risk stratification using the Framingham risk score (FRS) in a community-based, multiethnic population. Echocardiography and carotid ultrasonography were performed in 1,445 subjects (aged >39 years; 40% men; 53% Hispanic, 20% white, 24% black) from the Northern Manhattan Study. Subclinical cardiovascular disease was defined as the presence of left ventricular hypertrophy and/or carotid plaque greater than the gender-specific 75th percentile of the left ventricular mass index and maximal carotid plaque thickness distribution. The prevalence of subclinical cardiovascular disease was examined in each FRS category (low, intermediate, and high risk). In subjects with low or intermediate FRSs, 35% had subclinical cardiovascular disease (low FRS 29%, intermediate FRS 42%). In the intermediate FRS category, subclinical cardiovascular disease was significantly more prevalent in women than in men (53% vs 32%, p <0.0001) and in black and white subjects than in Hispanics (59% and 46% vs 33%, p <0.0001 and p = 0.040, respectively). In conclusion, the ultrasound assessment of subclinical cardiovascular disease may help reclassify 1/3 of subjects with low or intermediate FRSs into higher risk groups. In the intermediate FRS category, FRS appears to underestimate the coronary risk more in women than in men and more in whites and especially in blacks than in Hispanics. PMID- 17134633 TI - Repeated serum lipid measurements during the peri-hospitalization period. AB - The early treatment of hyperlipidemia in hospitalized patients confers potential benefit, yet total cholesterol is known to vary with acute illness, often delaying treatment decisions. A prospective study was conducted of 61 patients (mean age 57 years; 49% women) admitted to an acute care community hospital with various diagnoses with random nonfasting lipid profile measurements at admission, followed by second fasting lipid profile measurements on day 3 of hospitalization or upon discharge (whichever occurred first), and final fasting lipid profile measurements 4 weeks after discharge. All individual values of the lipid profile decreases at discharge, whereas the ratios of total cholesterol to high-density lipoprotein (HDL) and of low-density lipoprotein (LDL) to HDL did not change significantly. The 95% confidence interval around the total cholesterol/HDL ratio for each patient was within the same National Cholesterol Education Program Adult Treatment Panel III treatment recommendation category 42.6% of the time, whereas corresponding intervals for total cholesterol and LDL were within a single treatment category only 6.6% of the time. The total cholesterol/HDL ratio was significantly more consistent with regard to treatment implications than LDL or total cholesterol (p <0.0001). In conclusion, serum lipid values vary significantly during and after a hospital stay, whereas the ratio of total cholesterol to HDL remains relatively stable. This ratio may therefore serve as a more reliable basis for early treatment decisions in dyslipidemia. PMID- 17134634 TI - Effectiveness of the implantable cardioverter defibrillator in blacks versus whites (from MADIT-II). AB - The aim of this study was to examine the effectiveness of implantable cardioverter-defibrillator (ICD) therapy in whites and blacks who were enrolled in the Multicenter Automatic Defibrillator Implantation Trial-II (MADIT-II). The study population involved 1,232 subjects, with 1,073 white patients (87%) and 102 black patients (8%). Univariate analyses revealed a significant reduction in mortality in white patients treated with ICD therapy (p <0.02), but not in black patients (p = 0.96). White patients had a significant reduction of sudden cardiac death (p <0.01), but no benefit was evident in black patients (p = 0.62). Adjusting for relevant covariates, the ICD therapy/conventional therapy hazard ratios for total mortality were favorable in whites (0.75, 95% confidence interval [CI] 0.55 to 1.02), but not in blacks (1.25, 95% CI 0.42 to 3.60); the hazard ratios for sudden cardiac death were significantly (p = 0.04) lower in whites (0.29, 95% CI 0.17 to 0.49) than in blacks (1.71, 95% CI 0.33 to 8.84). Interaction analyses revealed a pattern of progressive improvement in ICD efficacy in whites as the end point became more specific for arrhythmic death, with exactly the reverse for blacks. In conclusion, ICD therapy in MADIT-II was associated with a reduction in total mortality, cardiac death, and sudden cardiac death in whites but not in blacks. PMID- 17134635 TI - Relation of arterial stiffness to left ventricular diastolic function and cardiovascular risk prediction in patients > or =65 years of age. AB - There is a paucity of data regarding the relation between the various noninvasive indexes of arterial stiffness and left ventricular diastolic function. In 188 subjects aged > or =65 years (mean 75 +/- 5; 71% men), the concordance and strength of the association between measures of arterial stiffness and left ventricular diastolic function were evaluated. Indexes of arterial stiffness (brachial and aortic pulse pressure [PP], carotid-femoral pulse-wave velocity [PWV], and augmentation pressure [AP]) were measured using applanation tonometry. Diastolic function was classified in terms of instantaneous diastolic function grade and quantitated as left atrial volume, a measure of chronic diastolic burden. Risk for new cardiovascular events was estimated using a validated clinical echocardiographic risk algorithm. Aortic and brachial PP, PWV, and AP were correlated positively with left atrial volume and diastolic function grade. After adjusting for age, gender, and clinical and echocardiographic covariates, 1 SD increases in aortic PP, brachial PP, PWV, and AP were associated with 6%, 6%, 4%, and 4% increases in indexed left atrial volume, respectively. Similarly, 1-SD increases in aortic PP, brachial PP, and AP were associated with 84%, 81%, and 83% increased risk for diastolic dysfunction, respectively (all p <0.04). PWV and aortic and brachial PP were superior to AP in discriminating subjects with the highest risk of having new cardiovascular events (5-year risk >50%; area under receiver-operating characteristic curve 0.67, 0.67, 0.70, and 0.56, respectively; p <0.05). In conclusion, increased arterial stiffness was associated with more severe left ventricular diastolic dysfunction, although the strength of the association varied according to the specific measure used. Aortic PP, brachial PP, and PWV appeared superior to AP in risk discrimination in this elderly cohort. PMID- 17134636 TI - Effects of omega-3 fatty acids on heart rate in cardiac transplant recipients. AB - Omega-3 fatty acid (n-3 FA) consumption has been linked to reductions in the risk of death from coronary heart disease and, recently, to lower heart rates (HRs). The investigators previously observed a reduction of 5 beats/min in HR in patients with coronary heart disease given n-3 FAs (eicosapentaenoic acid and docosahexaenoic acid) for 4 months. Reductions in HR may be explained by enhanced vagal tone, influences on cardiac voltage-gated ion channels, or both. The hypothesis that n-3 FAs would affect HR independent of vagal input was investigated by studying the effects of n-3 FA supplementation on HR in patients with denervated hearts after orthotopic heart transplantation. Electrocardiographic data obtained in 2 prospective trials in which 18 heart transplant recipients received n-3 FA supplementation (1 to 3.4 g/day) for 4 to 6 months were collected. Patients were studied 4.4 +/- 2.6 years after transplantation. HR, QRS complex, and QTc duration were determined before and after treatment. Pretreatment HR was reduced from 88 +/- 14 to 83 +/- 13 beats/min after n-3 FA treatment (p = 0.016). QRS duration increased from 107 +/- 24 to 117 +/- 25 ms (p = 0.001). QTc duration remained unchanged from baseline (427 +/- 34 ms) to study end (424 +/- 39 ms). In conclusion, n-3 FA supplementation reduced HR and prolonged QRS duration in heart transplant recipients who were presumably devoid of vagal innervation. These findings suggest that n-3 FAs may modify electrophysiologic properties of the myocardium itself. PMID- 17134637 TI - Left ventricular geometry and mortality in patients >70 years of age with normal ejection fraction. AB - Numerous studies have indicated that left ventricular (LV) hypertrophy predicts morbidity and mortality, including studies in older patients. The prognostic impact of other LV geometric patterns, including concentric remodeling (CR), compared with LV hypertrophy has not been studied in a large cohort of older patients. Echocardiographic and clinical data were studied in 9,771 consecutive patients aged >70 years with ejection fractions > or =50% who were followed for 3.1 years to determine the impact of LV geometric patterns. CR was the most prevalent pattern (43%), and 16% met criteria for LV hypertrophy. Although patients with CR had significantly lower LV mass indexes than patients with normal structure as well as those with LV hypertrophy, their mortality was 35% higher than that of normal subjects (15.5% vs 11.5%, p <0.0001) and 13% higher than that of patients with eccentric LV hypertrophy (15.5% vs 13.7%, p <0.001) with similar mortality to those with concentric LV hypertrophy (15.5% vs 15.9%). In conclusion, abnormal LV geometry is extremely common in patients aged >70 years with normal systolic function, being present in 59% of this population, with CR (43%) by far the most common LV geometric pattern. Patients aged >70 years with CR have significantly increased mortality compared with similarly aged patients with either normal structure or eccentric LV hypertrophy, with similar increased mortality to those with concentric LV hypertrophy. PMID- 17134638 TI - Characterization of the coronary sinus ostium by cardiac magnetic resonance imaging. AB - The diameter and the angle of the coronary sinus (CS) ostium was analyzed in 101 patients who underwent cardiac magnetic resonance imaging and had left ventricular ejection fractions < or =0.35 (n = 40) or > or =0.65 (n = 61). The angle of the CS ostium in patients with LVEFs < or =0.35 was less acute than in patients with LVEFs > or =0.65 (73 degrees +/- 12 degrees vs 65 degrees +/- 10 degrees, p <0.01). There was no statistically significant difference in the diameter of the CS ostium in patients with LVEFs < or =0.35 compared with those with LVEFs > or =0.65 (8 +/- 3 vs 8 +/- 2 mm, p = 0.5). The diameter and the angle of the CS ostium were not different when analyzed on the basis of the duration of the QRS complex, left atrial dimension, or left ventricular end diastolic dimension. In conclusion, on the basis of cardiac magnetic resonance imaging data, the angle of the CS is less acute in patients with LVEFs < or =0.35 than in those with LVEFs > or =0.65. PMID- 17134640 TI - Identifying patients for aggressive cholesterol lowering: the risk curve concept. AB - The National Cholesterol Education Program's 2004 report identified more aggressive optional low-density lipoprotein (LDL) cholesterol treatment goals of <70 mg/dl for secondary prevention patients and <100 mg/dl for moderately high risk, primary prevention patients. Although LDL cholesterol reduction is the first step in reducing cardiovascular risk, it may be difficult for clinicians to visualize the risk reduction benefit for patients from various risk interventions. The concept of a "risk curve," or the absolute risk of a patient for subsequent cardiovascular events over a range of LDL cholesterol values, is proposed. In conclusion, placing a patient on the appropriate risk curve may facilitate an individualized clinical management strategy that takes into account the patient's absolute benefit from further LDL cholesterol reduction as well as from shifting the risk curve downward through non-LDL cholesterol interventions. PMID- 17134641 TI - Costs and quality-of-life effects of implantable cardioverter-defibrillators. AB - Although the clinical efficacy of implantable cardioverter-defibrillators (ICDs) has been convincingly demonstrated in clinical trials, the impact of ICDs on health care costs and recipients' quality of life (QOL) is less certain. The existing medical research on the health care costs and QOL effects of ICDs was reviewed and summarized. Medline and the Institute for Scientific Information's Web of Knowledge were searched for publications reporting costs of care and QOL assessments of ICD recipients. Unpublished and non-peer-reviewed "gray" publications were excluded. Reports were included if they reported primary, original patient data that were collected after 1993, when nonthoracotomy defibrillators entered clinical practice. Two reviewers independently evaluated publications for relevance and quality, abstracted study data, and summarized the findings. Excessive heterogeneity among studies prevented formal meta-analysis, so a narrative synthesis was performed, and key themes were identified from the published research. There were limited published data on the costs of ICD care, especially for the primary prevention of sudden cardiac death. The published research on ICD QOL lacked large, multicenter, longitudinal studies. Many ICD QOL studies were performed in small numbers of patients at single centers. Initial ICD implantation costs ranged (in 2006 United States dollars) from $28,500 to $55,200, with annual follow-up costs ranging from $4,800 to $17,000. QOL was higher for ICD recipients than for patients treated with antiarrhythmic drugs, but there was a substantial prevalence of anxiety, depression, and "loss of control" in ICD recipients, particularly in those who had received ICD shocks. In conclusion, ICD implantation remains costly but may be becoming less expensive over time, and ICD recipients' QOL is significantly affected by their devices. PMID- 17134642 TI - Dose-dependent association between use of loop diuretics and mortality in advanced systolic heart failure. PMID- 17134643 TI - The role of platelets in ST-segment elevation myocardial infarction. PMID- 17134644 TI - Crystalline retinopathies. AB - Many types of crystalline retinopathies have been described, associated with a myriad of medical conditions ranging from chronic retinal conditions to inherited systemic diseases. This comprehensive review summarizes the different types of crystalline retinopathies, including their clinical presentations, diagnostic criteria, pathology, and treatment options. PMID- 17134645 TI - Madarosis. AB - Madarosis may be a presenting feature of a number of vision and life-threatening conditions, including herpes zoster, leprosy, HIV/AIDS, trachoma, malignant eyelid tumors, discoid lupus, scleroderma, and hypothyroidism. It may occur via two broad pathogenic pathways: scarring and non-scarring, which indicates the potential for lash re-growth. Madarosis may occur as an isolated finding or together with loss of other body and scalp hair. The etiology of madarosis can be further divided into dermatological, infection, endocrine, neoplastic, drug related, congenital, and trauma. This report includes salient points in the clinical history and examination of patients with madarosis, with an emphasis on excluding or diagnosing visual or life threatening disorders associated with madarosis. PMID- 17134646 TI - Current concepts of ocular manifestations in Marfan syndrome. AB - Marfan syndrome is a widespread disorder of connective tissue. It is characterized by systemic and ocular features due to mutations in the fibrillin gene. Awareness and prompt recognition of the ocular complications of Marfan syndrome may enable improvement and preservation of sight. Studies have been performed in the last few years that enable a better understanding of the genetics of the syndrome, earlier diagnosis, and improvement in the surgical techniques and options. PMID- 17134647 TI - Unraveling a complex genetic disease: age-related macular degeneration. AB - In most of the Western world, age-related macular degeneration (AMD) remains the largest single cause of severe visual impairment, and its prevalence continues to increase. It is considered to be a complex disease, in which multiple genes and environment play a role in pathogenesis. Several environmental insults are implicated with smoking, serum cholesterol, hypertension, sunlight exposure, and many other factors being variously associated with disease pathogenesis. Until recently, there have been relatively few breakthroughs to further our understanding of the genetics of AMD, despite remarkable progress in molecular genetic techniques over the last 20 years, and the fact that many rare inherited macular diseases have had their causative genes mapped. Development of new tools such as high-density single-nucleotide polymorphism chips and microarrays have changed the face of genetic research, but have yet to directly translate into improved clinical outcomes in ophthalmology. However with the recent finding of the Tyr402His polymorphism in the complement factor H gene being implicated in AMD, we are about to witness a new wave of research in this disease. Not only does the identification of a biologically plausible gene identify a new pathway, but it also identifies new biological mechanisms for disease, avenues to pursue treatment, and a better understanding of how the environment interacts with the genetic background to create disease. This article aims to review the process of gene discovery in complex disease, why the search for genes remains difficult, how to translate laboratory findings to a clinical setting, and how these findings will impact on disease treatment and public health issues. PMID- 17134648 TI - Double vision worth a double take. AB - An 18-year-old girl presented with an acute acquired comitant esotropia. Investigation showed a brainstem glioma. Discussion of types of acute acquired comitant esotropia, differential diagnosis and neurologic work-up is given. Review of the literature involving acute acquired comitant esotropia is provided. PMID- 17134649 TI - The woman who needed a pet. AB - A 72-year-old woman developed difficulty reading, driving, and playing dominoes. Ophthalmologic examination revealed a homonymous hemianopia, but brain MRI showed no abnormality to explain the visual field defect. Neuropsychiatric testing demonstrated severely impaired visual processing (simultagnosia, visual agnosia, visuospatial difficulty). Positron emission tomography revealed hypometabolism of both parietal and occipital lobes consistent with posterior cortical atrophy or the visual variant of Alzheimer disease. Functional neuroimaging should be considered in the setting of a normal MRI among patients with signs and symptoms of the visual variant Alzheimer disease. PMID- 17134650 TI - The eye disease of Jefferson Davis (1808-1889). AB - The only Confederate president, Jefferson Davis, led a long and eventful life. He was a Mississippi planter, a husband, a father, West Point graduate, war hero, congressman, senator, secretary of war, and finally President of the Confederate States of America. In many ways he was a study of contrast with his northern counterpart, Abraham Lincoln. Davis was personally courageous and a rich, educated, southern aristocrat who did not deeply understand the political process or have the refined personal skills necessary to work well with others. Prior to his Presidency he served with distinction in two wars, but as a result of his confederate activity and pro-slavery philosophy he is one of the least discussed famous Americans. Davis's health was a constant problem and he suffered an almost fatal attack of "malaria" in 1836. In the winter of 1857-1858 he again was seriously ill and by the end of February 1858 a chronic, relapsing, ocular inflammatory condition began. Using historical evidence from multiple sources, this paper will propose a diagnosis of the Confederate President's ocular condition and consider how this could have influenced his military and political decisions. PMID- 17134652 TI - Phylogenetic analyses of parasites in the new millennium. AB - Phylogenetic analysis has changed greatly in the last decade, and the most important themes in that change are reviewed here. Sequence data have become the most common source of phylogenetic information. This means that explicit models for evolutionary processes have been developed in a likelihood context, which allow more realistic data analyses. These models are becoming increasingly complex, both for nucleotides and for amino acid sequences, and so all such models need to be quantitatively assessed for each data set, to find the most appropriate one for use in any particular tree-building analysis. Bayesian analysis has been developed for tree-building and is greatly increasing in popularity. This is because a good heuristic strategy exists, which allows large data sets to be analyzed with complex evolutionary models in a practical time. Perhaps the most disappointing aspect of tree interpretation is the ongoing confusion between rooted and unrooted trees, while the effect of taxon and character sampling is often overlooked when constructing a phylogeny (especially in parasitology). The review finishes with a detailed consideration of the analysis of a multi-gene data set for several dozen taxa of Cryptosporidium (Apicomplexa), illustrating many of the theoretical and practical points highlighted in the review. PMID- 17134653 TI - Targeting of toxic compounds to the trypanosome's interior. AB - Drugs can be targeted into African trypanosomes by exploiting carrier proteins at the surface of these parasites. This has been clearly demonstrated in the case of the melamine-based arsenical and the diamidine classes of drug that are already in use in the treatment of human African trypanosomiasis. These drugs can enter via an aminopurine transporter, termed P2, encoded by the TbAT1 gene. Other toxic compounds have also been designed to enter via this transporter. Some of these compounds enter almost exclusively through the P2 transporter, and hence loss of the P2 transporter leads to significant resistance to these particular compounds. It now appears, however, that some diamidines and melaminophenylarsenicals may also be taken up by other routes (of yet unknown function). These too may be exploited to target new drugs into trypanosomes. Additional purine nucleoside and nucleobase transporters have also been subverted to deliver toxic agents to trypanosomes. Glucose and amino acid transporters too have been investigated with a view to manipulating them to carry toxins into Trypanosoma brucei, and recent work has demonstrated that aquaglyceroporins may also have considerable potential for drug-targeting. Transporters, including those that carry lipids and vitamins such as folate and other pterins also deserve more attention in this regard. Some drugs, for example suramin, appear to enter via routes other than plasma-membrane mediated transport. Receptor-mediated endocytosis has been proposed as a possible way in for suramin. Endocytosis also appears to be crucial in targeting natural trypanocides, such as trypanosome lytic factor (TLF) (apolipoprotein L1), into trypanosomes and this offers an alternative means of selectively targeting toxins to the trypanosome's interior. Other compounds may be induced to enter by increasing their capacity to diffuse over cell membranes; in this case depending exclusively on selective activity within the cell rather than selective uptake to impart selective toxicity. This review outlines studies that have aimed to exploit trypanosome nutrient uptake routes to selectively carry toxins into these parasites. PMID- 17134654 TI - Making sense of the schistosome surface. AB - The syncytial cytoplasmic layer, termed the tegument, which covers the entire surface of adult schistosomes, is a major interface between the parasite and its host. Since schistosomes can survive for decades within the host bloodstream, they are clearly able to evade host immune responses, and their ability is dependent on the properties of the tegument surface. We review here the molecular organization and biochemical functions of the tegument, combining the extensive literature over the last three decades with recent proteomic studies. We have interpreted the organization of the tegument surface as bounded by a conventional plasma membrane overlain by a membrane-like secretion, the membranocalyx, with which host molecules can associate. The range of parasite proteins, glycans and lipids found in the surface complex is evaluated, together with the host molecules detected. We consider the way in which the tegument surface is formed after cercarial penetration into the skin, and changes that occur as parasites develop to maturity. Lastly, we review the evidence on surface dynamics and turnover. PMID- 17134655 TI - Immunology and pathology of intestinal trematodes in their definitive hosts. AB - This review examines the significant literature on the immunology and pathology of intestinal trematodes in their definitive hosts. We emphasize information on selected species in six families for which the literature on these topics is extensive. The families are Brachylaimidae, Diplostomidae, Echinostomatidae, Gymnophallidae, Heterophyidae, and Paramphistomidae. For most of these families, coverage is considered under the following headings: (i) background; (ii) pathology of the infection; (iii) immunology of the infection; (iv) immunodiagnosis; and (v) human infection. Some of these heading have been subdivided further, based on the literature available on a particular topic. Following this coverage, we include a final section on the important topical literature on selected trematodes in families other than the six mentioned above. PMID- 17134656 TI - Systematics and epidemiology of trichinella. AB - In this review, we describe the current knowledge on the systematics, ecology and epidemiology of Trichinella and trichinellosis, and the impact of recent research discoveries on the understanding of this zoonosis. The epidemiology of this zoonosis has experienced important changes over the past two decades, especially with regard to the importance of the sylvatic cycle and the sylvatic species. Outbreaks of trichinellosis due to Trichinella spiralis from domestic swine, while still frequent, increasingly are caused by other Trichinella spp. infecting hosts such as horses, dogs, wild boars, bears and walruses. The latter revelations have occurred as a result of a series of discoveries on the systematics of Trichinella spp., facilitated by new molecular tools. As a consequence, the genus is now composed of two clades, an encapsulated group (five species and three genotypes) and a non-encapsulated one (three species). This has sparked renewed investigations on the host range of these parasites and their epidemiological features. Most dramatic, perhaps, is the recognition that reptiles may also serve as hosts for certain species. This new knowledge base, in addition to having an important relevance for food safety policies and protection measures, is raising important questions on the phylogeny of Trichinella spp., the ecological characteristics of the species and their geographic histories. Answers to these questions may have great value for the understanding of the evolutionary biology for other parasitic helminths, and may increase the value of this genus as models for research on parasitism in general. PMID- 17134657 TI - The importance of public health: let history speak for itself. PMID- 17134658 TI - The interactive effect of alcohol and nicotine on NGF-treated pheochromocytoma cells. AB - Previous studies have reported that alcohol exposure reduces the number of neuronal-like pheochromocytoma (PC12) cells in culture. In this study, the interactive effect of coexposure of alcohol and nicotine on PC12 cell numbers was examined in comparison with the effect derived from alcohol or nicotine exposure individually. Moreover, the role of apoptosis in mediating changes in PC12 cell numbers was also investigated. It was hypothesized that alcohol would result in cell loss, and the presence of nicotine would attenuate the damaging effects of alcohol. PC12 cells were exposed to alcohol (100 mM), nicotine (10 microM), or both alcohol and nicotine for 24, 48, 72, or 96 h. Caspase-3 activity and DNA fragmentation, markers for apoptotic cell death, were measured to determine the role of apoptosis in mediating decreases in PC12 cell numbers. The findings indicated that both alcohol and nicotine exposure significantly decreased PC12 cell numbers when compared with the control treatment. Furthermore, the coexposure of these two drugs caused a significantly greater decrease in cell numbers when compared with cells exposed to either alcohol or nicotine alone. This additive effect was related to the duration of exposure with a marked reduction in cell numbers following 96 h of coexposure to alcohol and nicotine. Neither alcohol nor nicotine exposure appeared to alter caspase-3 activity or DNA fragmentation levels, suggesting that the reduction in PC12 cell numbers following alcohol and/or nicotine exposure may possibly be due to factors other than apoptosis, such as interference with proliferation rates. PMID- 17134659 TI - Association of alcohol-metabolizing genes with alcoholism in a Mexican Indian (Otomi) population. AB - Association studies provide a powerful approach to link DNA variants and genetic predisposition to complex diseases. In this study, we determined the genotype and allelic frequencies of genes encoding enzymes involved in alcohol metabolism in alcoholic and nonalcoholic subjects of related ethnicity. A total of 118 individuals of Otomi Mexican Indian ancestry were included. Fifty-nine were chronic alcoholics according to WHO criteria and alcohol dependents according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM IV) criteria. They were compared to 59 teetotalers or alcohol consumers of <10 g per day. The restriction fragment length polymorphisms analyzed were ADH1B/MaeIII, ALDH2/MboII, CYP2E1/DraI, CYP2E1/RsaI, and CYP2E1/TaqI. Of the studied polymorphisms, a significant difference between alcoholic and nonalcoholic Otomies was observed only in the CYP2E1/TaqI. The common genotype in alcoholics was A1/A2 (54%), and in nonalcoholics the homozygous A2/A2 (63%) (odds ratio [OR]: 0.28; 95% confidence interval [CI]: 0.13-0.60; P=.002). The frequency of the mutant allele A1 was significantly higher in alcoholics than in nonalcoholics (41 vs. 21%; OR: 2.4; 95% CI: 1.3-4.3; P=.003). This documents the presence of a polymorphism of CYP2E1 that is overexpressed in alcoholic Otomies, in which the variant allele (A1 of CYP2E1/TaqI) is associated with increased susceptibility to alcoholism. The appreciation that this finding may be an additional factor contributing to the high frequency of liver cirrhosis in Otomies requires further investigation. PMID- 17134660 TI - ADH1C*2 allele is associated with alcohol dependence and elevated liver enzymes in Trinidad and Tobago. AB - Variants in alcohol dehydrogenase (ADH) genes differ between ethnic groups and have, in some studies, been found to be associated with alcohol dependence and alcoholic liver disease. This study sought to determine whether an association exists between ADH (ADH1C previously ADH3, ADH1B*2 previously ADH2*2) genotypes, alcohol dependence, drinking history, and liver function tests in the two major ethnic groups of Trinidad and Tobago (TT). One hundred and forty-five alcohol dependent individuals of both East Indian (Indo-TT) and African (Afro-TT) ancestry, and 108 controls matched by age, sex, and education participated in the study. Serum levels of alanine and aspartate aminotransferase (ALT, AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and gamma glutamyltransferase (GGT) as well as presence of HIV, hepatitis B surface antigen, and anti-hepatitis C virus antibody were determined. There was a significant difference in the distribution of ADH1C allele polymorphisms between the ethnic groups (P<.0001). Forty-three percent of the Indo-TT were found to have one ADH1C*2 allele and 5% were homozygous, whereas, only 23% of Afro-TT had one allele and one was homozygous. Only three individuals had an ADH1B*2 allele (one Indo-TT alcohol dependent, two Indo-TT controls). The ADH1C*2 allele was significantly associated with alcohol dependence overall and within Indo-TT ancestry, however, it was not associated with current or heaviest alcohol consumption levels. Individuals with at least one ADH1C*2 allele also had significantly elevated levels of ALP (P<.02) and GGT (P<.02) as compared to individuals homozygous for ADH1C*. Additionally, GGT levels were also found to be elevated (P<.02) within Indo-TT alcohol dependents with at least one ADH1C*2 allele but not within the Afro-TT alcohol dependents with that allele. A linear regression that included alcohol dependence and levels of alcohol consumption confirmed that levels of serum GGT were significantly associated with the ADH1C*2 genotype. These results suggest that ADH1C polymorphisms are associated with alcohol dependence and alcohol associated elevations of liver enzymes in a population with a low frequency of ADH1B2 alleles. PMID- 17134661 TI - A comparative survey on alcohol and tobacco use in urban and rural populations in the Huaihua District of Hunan Province, China. AB - Modern Chinese society is comprised primarily of two subgroups-urban and rural subpopulations. However, comparative data regarding alcohol and tobacco use between urban and rural subgroups in China is sparse. An epidemiological survey was conducted in the Huaihua District of Hunan Province of China, and 3,543 urban and 4,294 rural dwellers aged 15-65 years were interviewed. The drinking rates were higher in the urban area (45.9%) than in the rural area (39.6%), whereas the smoking rates were higher in the rural area (35.9%) than in the urban area (28.7%). Rural respondents were more likely to report heavy drinking (Adjusted odds ratio [OR]=1.77) and heavy smoking (Adjusted OR=2.46) than urban populations. Specifically, rural males had higher odds of heavy alcohol and tobacco use than urban males, whereas more urban females were more likely to drink and smoke than rural counterparts. The findings suggest that the status of alcohol and tobacco use varies in the urban and rural area. The heavy alcohol and tobacco consumption among rural populations underscores the need to develop and implement culturally appropriate public health intervention and awareness arising programs especially in rural China. PMID- 17134662 TI - The effect of acetaldehyde-glycosaminoglycan mixtures upon Factor IXa and Factor IX-Deficient Plasma. AB - Earlier studies have shown that acetaldehyde, the primary intermediate in the biological degradation of ethanol, interacts with enzymes and zymogens of the common coagulation pathway, prolonging prothrombin time and activated partial thromboplastin time (APTT), and that acetaldehyde-glycosaminoglycans (GAGs) mixtures synergistically prolong clotting times (Brecher, A. S. (2005). In Comprehensive Handbook of Alcohol Related Pathology. Vol. 3(93), pp. 1223-1244). In this study, the effect of acetaldehyde and GAGs upon Factor IXa, an intrinsic pathway enzyme, has been investigated. Individually, acetaldehyde, heparins of various molecular weights, dermatan sulfate, and chondroitin sulfates A and C affect Factor IXa, prolonging clotting time as measured by APTT. Pre-incubation of Factor IXa with a mixture of 22.3 mM acetaldehyde and heparin(17k), heparin(6k), dermatan sulfate, or chondroitin sulfate A additively prolongs clotting times, reflecting individual, unrelated molecular mechanistic effects. In contrast, a synergistic effect is observed at the 44.7 mM acetaldehyde level with heparin(17k), heparin(3k), chondroitin sulfates A and C, and dermatan sulfate, suggesting that acetaldehyde may cross-link with the enzyme and the GAGs, forming tertiary complexes, further influencing coagulopathy. These observations upon Factor IXa present a deeper dimension to the anticoagulation effect of alcohol on the coagulation cascade. PMID- 17134663 TI - An explanation for the paradoxical induction and suppression of an acute phase response by ethanol. AB - Binge ethanol (EtOH) consumption suppresses inflammatory responses and resistance to infection, but paradoxically it is associated with increased levels of acute phase proteins (which are indicators of inflammation) and an increased risk of inflammation-mediated pathologies such as cardiovascular disease and cirrhosis of the liver. The latter effect may be mediated by increased translocation of bacteria leading to activation of toll-like receptor 4 (TLR4). In this study, the dose-response and time course of the effects of EtOH alone or EtOH in conjunction with a TLR4 agonist (lipopolysaccharide [LPS]) were evaluated in mice. EtOH alone at a dosage of 6 g/kg induced an acute phase response (as indicated by enzyme linked immunosorbent assay for serum amyloid A and serum amyloid P) that was maximal 24 h after dosing. Lower dosages of EtOH did not have this effect but did suppress the acute phase response to LPS and the production of interleukin-6 up to 3 h after dosing. EtOH at 6 g/kg did not induce an acute phase response in C3H/HeJ (TLR4 mutant) mice, indicating that this response is mediated through TLR4. These results provide a resolution for the apparently paradoxical pro- and anti-inflammatory actions of EtOH with regard to acute phase responses. PMID- 17134664 TI - Study on surface dose generated in prostate intensity-modulated radiation therapy treatment. AB - The surface doses of 6- and 15-MV prostate intensity-modulated radiation therapy (IMRT) irradiations were measured and compared to those from a 15-MV prostate 4 beam box (FBB). IMRT plans (step-and-shoot technique) using 5, 7, and 9 beams with 6- and 15-MV photon beams were generated from a Pinnacle treatment planning system (version 6) using computed tomography (CT) scans from a Rando Phantom (ICRU Report 48). Metal oxide semiconductor field effect transistor detectors were used and placed on a transverse contour line along the Phantom surface at the central beam axis in the measurement. Our objectives were to investigate: (1) the contribution of the dynamic multileaf collimator (MLC) to the surface dose during the IMRT irradiation; (2) the effects of photon beam energy and number of beams used in the IMRT plan on the surface dose. The results showed that with the same number of beams used in the IMRT plan, the 6-MV irradiation gave more surface dose than that of 15 MV to the phantom. However, when the number of beams in the plan was increased, the surface dose difference between the above 2 photon energies became less. The average surface dose of the 15-MV IMRT irradiation increased with the number of beams in the plan, from 0.86% to 1.19%. Conversely, for 6 MV, the surface dose decreased from 1.33% to 1.24% as the beam number increased from 7 to 9. Comparing the 15-MV FBB and 6-MV IMRT plans with 2 Gy/fraction, the IMRT irradiations gave generally more surface dose, from 15% to 30%, depending on the number of beams in the plan. It was found that the increase in surface dose for the IMRT technique compared to the FBB plan was predominantly due to the number of beams and the calculated monitor units required to deliver the same dose at the isocenter in the plans. The head variation due to the dynamic MLC movement changing the surface dose distribution on the patient was reflected by the IMRT dose-intensity map. Although prostate IMRT in this study had an average higher surface dose than that of FBB, the more even distribution of relatively lower surface dose in IMRT field could avoid the big dose peaks at the surface positions directly under the FBB fields. Such an even and low surface dose distribution surrounding the patient in IMRT is believed to give less skin complication than that of FBB with the same prescribed dose. PMID- 17134665 TI - Inverse planning of energy-modulated electron beams in radiotherapy. AB - The use of megavoltage electron beams often poses a clinical challenge in that the planning target volume (PTV) is anterior to other radiosensitive structures and has variable depth. To ensure that skin as well as the deepest extent of the PTV receives the prescribed dose entails prescribing to a point beyond the depth of peak dose for a single electron energy. This causes dose inhomogeneities and heightened potential for tissue fibrosis, scarring, and possible soft tissue necrosis. Use of bolus on the skin improves the entrant dose at the cost of decreasing the therapeutic depth that can be treated. Selection of a higher energy to improve dose homogeneity results in increased dose to structures beyond the PTV, as well as enlargement of the volume receiving heightened dose. Measured electron data from a linear accelerator was used as input to create an inverse planning tool employing energy and intensity modulation using bolus (e-IMRT). Using tools readily available in a radiotherapy department, the applications of energy and intensity modulation on the central axis makes it possible to remove hot spots of 115% or more over the depths clinically encountered. The e-IMRT algorithm enables the development of patient-specific dose distributions with user-defined positions of peak dose, range, and reduced dose to points beyond the prescription point. PMID- 17134666 TI - A multiple acquisition sequence for IMRT verification with a 2D ion chamber array. AB - This investigation deals with the implementation of a multiple acquisition (MA) sequence for radiotherapy beam verification with a 2-dimensional (2D) ion chamber array. The acquisition is carried out through multiple beam deliveries with the 2D detector array in different positions. The MA sequence is performed through remote-controlled movements of the treatment couch. In this work, the MA dose map of an intensity-modulated radiotherapy (IMRT) modulated beam is presented and compared to standard acquisition mode data and ion chamber measurements. The implementation of MA sequence increases the number of measurement points and therefore the efficiency of the detection system and the final imaging resolution. Results from this investigation show that the imaging resolution of the system used in standard acquisition mode can be increased up to 4 times. The procedure described in this work can be automated, including couch movements in the radiotherapy plan sent to the treatment workstation. Furthermore, this specific solution could be successfully applied to matrices of detector with a different construction design. PMID- 17134667 TI - Comparison of Kodak EDR2 and Gafchromic EBT film for intensity-modulated radiation therapy dose distribution verification. AB - The quantitative dose validation of intensity-modulated radiation therapy (IMRT) plans require 2-dimensional (2D) high-resolution dosimetry systems with uniform response over its sensitive region. The present work deals with clinical use of commercially available self-developing Radio Chromic Film, Gafchromic EBT film, for IMRT dose verification. Dose response curves were generated for the films using a VXR-16 film scanner. The results obtained with EBT films were compared with the results of Kodak extended dose range 2 (EDR2) films. The EBT film had a linear response between the dose range of 0 to 600 cGy. The dose-related characteristics of the EBT film, such as post irradiation color growth with time, film uniformity, and effect of scanning orientation, were studied. There was up to 8.6% increase in the color density between 2 to 40 hours after irradiation. There was a considerable variation, up to 8.5%, in the film uniformity over its sensitive region. The quantitative differences between calculated and measured dose distributions were analyzed using DTA and Gamma index with the tolerance of 3% dose difference and 3-mm distance agreement. The EDR2 films showed consistent results with the calculated dose distributions, whereas the results obtained using EBT were inconsistent. The variation in the film uniformity limits the use of EBT film for conventional large-field IMRT verification. For IMRT of smaller field sizes (4.5 x 4.5 cm), the results obtained with EBT were comparable with results of EDR2 films. PMID- 17134668 TI - Inverse-planned, dynamic, multi-beam, intensity-modulated radiation therapy (IMRT): a promising technique when target volume is the left breast and internal mammary lymph nodes. AB - The purpose of this study was to determine the optimum beam number and orientation for inverse-planned, dynamic intensity-modulated radiation therapy (IMRT) for treatment of left-sided breast cancer and internal mammary nodes (IMNs) to improve target coverage while reducing cardiac and ipsilateral lung irradiation. Computed tomography (CT) data was used from 5 patients with left sided breast cancer in whom the heart was close to the chest wall. The planning target volume (PTV) was the full breast plus ipsilateral IMNs. Two geometric beam arrangements were investigated, 240 degrees and 190 degrees sector angles, and the number of beams was increased from 7 to 9 to 11. Dose comparison metrics included: PTV homogeneity and conformity indices (HI, CI), heart V30, left lung V20, and mean doses to surrounding structures. To assess clinical application, the IMRT plans with 11 beams equally spaced in a 190 degrees sector angle were compared to conventional plans. Treatment times were modeled. The 190 degrees IMRT plans improved PTV HI and CI and reduced mean dose to the heart, lungs, contralateral breast, and total healthy tissue (all p < 0.05) compared to a 240 degrees sector angle. The 11-beam plan significantly improved PTV HI and CI, heart V30, left lung V20, and healthy tissue V5 compared to a 7-beam plan (all p < 0.05). The 11-beam plan reduced heart V30 and left lung V20 (p < 0.05) without compromising PTV coverage, compared to a 9-beam plan. Compared to a conventional plan, the IMRT class solution significantly improved PTV HI and CI (both p < 0.01), heart V30 (p = 0.01), and marginally reduced left lung V20 (p = 0.07) but increased contralateral breast and lung mean dose (p < 0.001) and healthy tissue V5 (p < 0.001). An 11-beam 190 degrees sector angle IMRT technique as a class solution is clinically feasible. PMID- 17134669 TI - Intensity-modulated radiotherapy for neoadjuvant treatment of gastric cancer. AB - Radiation therapy plays an integral role in the treatment of gastric cancer in the postsurgery setting, the inoperable/palliative setting, and, as in the case of the current report, in the setting of neoadjuvant therapy prior to surgery. Typically, anterior-posterior/posterior-anterior (AP/PA) or 3-field techniques are used. In this report, we explore the use of intensity-modulated radiotherapy (IMRT) treatment in a patient whose care was transferred to our institution after 3-field radiotherapy (RT) was given to a dose of 30 Gy at an outside institution. If the 3-field plan were continued to 50 Gy, the volume of irradiated liver receiving greater than 30 Gy would have been unacceptably high. To deliver the final 20 Gy, an opposed parallel AP/PA plan and an IMRT plan were compared to the initial 3-field technique for coverage of the target volume as well as dose to the kidneys, liver, small bowel, and spinal cord. Comparison of the 3 treatment techniques to deliver the final 20 Gy revealed reduced median and maximum dose to the whole kidney with the IMRT plan. For this 20-Gy boost, the volume of irradiated liver was lower for both the IMRT plan and the AP/PA plan vs. the 3 field plan. Comparing the IMRT boost plan to the AP/PA boost-dose range (<10 Gy) in comparison to the AP/PA plan; however, the IMRT plan irradiated a smaller liver volume within the higher dose region (>10 Gy) in comparison to the AP/PA plan. The IMRT boost plan also irradiated a smaller volume of the small bowel compared to both the 3-field plan and the AP/PA plan, and also delivered lower dose to the spinal cord in comparison to the AP/PA plan. Comparison of the composite plans revealed reduced dose to the whole kidney using IMRT. The V20 for the whole kidney volume for the composite IMRT plan was 30% compared to approximately 60% for the composite AP/PA plan. Overall, the dose to the liver receiving greater than 30 Gy was lower for the composite IMRT plan and was well below acceptable limits. In conclusion, our study suggests a dosimetric benefit of IMRT over conventional planning, and suggests an important role for IMRT in the neoadjuvant treatment of gastric cancer. PMID- 17134670 TI - Effects of neurosurgical titanium mesh on radiation dose. AB - The purpose of this study was to determine the dosimetric impact of a neurosurgical titanium mesh in patients treated with 6- and 18-MV photon beams. The effects of a 0.4-mm-thick titanium mesh on the dose profile at 3 regions within a solid water phantom were measured using extended dose range-2 (EDR2) film for 6- and 18-MV photon beams. All measurements were performed with the titanium mesh placed at a depth of 1.5 cm in the phantom. Films were exposed immediately above the mesh, immediately below the mesh, and at a depth of 5 cm from the surface of the phantom. The films were scanned using a scanning densitometer. In the region directly above the titanium mesh, there was an increase in dose of 7.1% for 6-MV photons and 4.9% for 18-MV photons. Directly below the titanium mesh, there was an average decrease in dose of 1.5% for 6-MV photons and an increase of 1.0% for 18-MV photons. At 5-cm depth, for 6- and 18 MV photons, there was a decrease in dose of 2.2% and 0.6%, respectively. We concluded that for cranial irradiation with high-energy photons, the dosimetric impact of a 0.4-mm titanium mesh is small and does not require modification in treatment parameters. PMID- 17134671 TI - Determination of cell electroporation from the release of intracellular potassium ions. AB - When cells are exposed to a strong enough external electric field, transient aqueous pores are formed in the membrane. The fraction of electroporated cells can be determined by measuring the release of intracellular potassium ions. The current work is the first study where such a method was employed successfully not only with cells suspended in the medium with a rather high concentration of potassium (4-5 mM) but also with cells that release some part of intracellular potassium responding, in this way, to the stress caused by manipulation procedures during the preparation of the cell suspension. Experiments were carried out on mouse hepatoma MH-A22 cells exposed to a square-wave electric pulse. The curves showing the dependence of the fraction of the cells that have become permeable to bleomycin, a membrane-impermeable cytotoxic drug, are close to the ones showing the release of intracellular potassium ions. PMID- 17134672 TI - Disposable biosensor based on enzyme immobilized on Au-chitosan-modified indium tin oxide electrode with flow injection amperometric analysis. AB - Indium tin oxide (ITO) electrode is used to fabricate a novel disposable biosensor combined with flow injection analysis for the rapid determination of H2O2. The biosensor is prepared by entrapping horseradish peroxidase (HRP) enzyme in colloidal gold nanoparticle-modified chitosan membrane (Au-chitosan) to modify the ITO electrode. The biosensor is characterized by scanning electron microscope, atomic force microscope, and electrochemical methods. Parameters affecting the performance of the biosensor, including concentrations of o phenylenediamine (OPD) and pH of substrate solution, were optimized. Under the optimal experimental conditions, H2O2 could be determined in the linear calibration range from 0.01 to 0.5 mM with a correlation coefficient of 0.997 (n=8). The amperometric response of the biosensor did not show an obvious decrease after the substrates were injected continuously 34 times into the flow cell. The prepared biosensor not only is economic and disposable, due to the low cost ITO film electrode obtained from industrial mass production, but also is capable with good detection precision, acceptable accuracy, and storage stability for the fabrication in batch. PMID- 17134673 TI - Design of peptide substrates for nanosecond time-resolved fluorescence assays of proteases: 2,3-diazabicyclo[2.2.2]oct-2-ene as a noninvasive fluorophore. AB - Fluorescence protease assays were investigated with peptide substrates containing a 2,3-diazabicyclo[2.2.2]oct-2-ene-labeled asparagine (Dbo) as a fluorescent amino acid. The special characteristic of the fluorophore Dbo is its exceedingly long fluorescence lifetime (ca. 300 ns in water under air), which allows the use of nanosecond time-resolved fluorescence (Nano-TRF) detection to efficiently suppress shorter-lived background emission. In addition, the natural amino acids tryptophan and tyrosine can be employed as intramolecular fluorescence quenchers, which facilitates substrate design. Fourteen synthetic peptide substrates (composed of 2-19 amino acids) and five enzymes (trypsin, pepsin, carboxypeptidase A, leucine aminopeptidase, and chymotrypsin) were investigated and, in all 28 examined combinations, enzymatic activity was detected by monitoring the increase in steady state fluorescence with time and determining the reaction rates as kcat/Km values, which ranged from 0.2 to 80x10(6) M-1 min 1. The results suggest an excellent compatibility of the very small and hydrophilic fluorescent probe Dbo with solid-phase peptide synthesis and the investigated proteases. For all 14 peptides the fluorescence lifetimes before and after enzymatic cleavage were measured and Nano-TRF measurements were performed in 384-well microplates. The fluorescence lifetimes of the different peptides provide the basis for the rational design of Dbo-based fluorescent substrates for protease assays. Measurements in Nano-TRF mode revealed, in addition to efficient suppression of background fluorescence, an increased differentiation between cleaved and uncleaved substrate. The Dbo-based assays can be adapted for high throughput screening. PMID- 17134674 TI - Determination of carbon labeling distribution of intracellular metabolites from single fragment ions by ion chromatography tandem mass spectrometry. AB - Liquid chromatography tandem mass spectrometry coupling is a highly sensitive and specific technique allowing molecule detection in the femtomolar range. This article introduces a straightforward approach to apply this technique in 13C metabolic flux analysis. Based on a theoretical analysis of the correlation between molecule ions and corresponding fragments, a method was developed to determine the carbon labeling of intracellular metabolites without increasing the number of measurements per metabolite compared with direct molecule ion analysis. The method was applied to phosphorylated metabolites because their fragmentation results in high yields of [PO3]- and/or [H2PO4]- ions. Comparing the accuracy of the carbon labeling determination of phosphorylated metabolites between direct analysis of the molecule ions with that of corresponding phosphate fragment ions, it could be demonstrated that the introduced approach resulted in significantly higher accuracy and sensitivity for all tested metabolites. When applying the techniques to Escherichia coli cell extracts, 2 microg cell dry weight per injection was sufficient to determine the natural abundances of the carbon fractions m and m+1 from six phosphorylated metabolites with high accuracy, predestining the approach for very small cultivation volumes in the microliter range. PMID- 17134675 TI - A rapid assay for dihydropteroate synthase activity suitable for identification of inhibitors. AB - The enzymes 6-hydroxymethylpterin pyrophosphokinase (HPPK) and dihydropteroate synthase (DHPS) catalyze sequential steps in folate biosynthesis. They are present in microorganisms but absent in mammals and therefore are especially suitable targets for antimicrobials. Sulfa drugs (sulfonamides and sulfones) currently are used as antimicrobials targeting DHPS, although resistance to these drugs is increasing. The most widely used assay that measures activity of these enzymes, to assess new inhibitors in vitro, is not amenable to automation. This article describes a simple, coupled, enzymatic spectrophotometric assay where the product of the DHPS reaction, dihydropteroate, is reduced to tetrahydropteroate by excess dihydrofolate reductase (DHFR) using the cofactor NADPH. The oxidation of NADPH is monitored at 340 nm. The activity of both HPPK and DHPS can be measured in this assay, and it has been used to measure kinetic parameters of wild-type and sulfa drug-resistant DHPS enzymes to demonstrate the utility of the assay. It is a sensitive and reproducible assay that can be readily automated and used in multiwell plates. This NADPH-coupled microplate photometric assay could be used for rapid screening of chemical libraries for novel inhibitors of folate biosynthesis as the first step in developing new antimicrobial drugs targeting the folate biosynthetic pathway. PMID- 17134676 TI - Identification of peroxisome-proliferator responsive element in the mouse HSL gene. AB - Hormone-sensitive lipase (HSL) catalyzes the rate-limiting step of lipolysis in adipose tissue. Several studies suggest that protein phosphorylation regulates the HSL enzymatic activity. On the other hand, the precise mechanism of the transcriptional regulation of the HSL gene remains to be elucidated. Here, we identified a functional peroxisome-proliferator responsive element (PPRE) in the mouse HSL promoter by reporter assay in CV-1 cells using serial deletion and point mutants of the 5'-flanking region. Chromatin immunoprecipitation (ChIP) analysis revealed that both peroxisome-proliferator activated receptor (PPARgamma) and retinoid X receptor (RXRalpha) interacted with the region. Binding of the PPARgamma/RXRalpha heterodimer to the PPRE sequence was also confirmed by electrophoretic mobility shift assay. These results indicate that the HSL gene is transcriptionally regulated by PPARgamma/RXRalpha heterodimer, and suggest that a cis-acting element regulates the HSL gene expression. PMID- 17134677 TI - Heterodimerization of the alpha and beta isoforms of the human thromboxane receptor enhances isoprostane signaling. AB - Isoprostanes are free radical catalyzed products of arachidonic acid that are elevated in pro-oxidant disease states. Two isoprostanes, 8-isoprostaglandin F(2alpha) (iPF(2alpha)III) and 8-isoprostaglandin E2 (iPE2III), act at the receptor for thromboxane A2 (the TP) to mediate pro-atherogenic effects in vivo. We confirmed dimerization of the human TP isoforms, TPalpha and TPbeta, and determined the impact on isoprostane signaling. No overt changes in ligand binding at the TP were observed as a result of TPalpha/TPbeta coexpression. The response to iPF(2alpha)III or iPE2III was enhanced in HEK293 cells stably coexpressing TPalpha and TPbeta, as measured by inositol phosphate generation or intracellular calcium mobilization, relative to cells expressing TPalpha or TPbeta individually. In contrast, the response to traditional thromboxane analogs was unaltered. Augmented isoprostane signaling was similarly observed in HEK 293 cell transiently transfected with TPalpha and TPbeta. These results indicate that TPalpha/TPbeta dimerization enhances isoprostane-mediated signal transduction. PMID- 17134678 TI - Nascent chain, mRNA, and ribosome complexes generated by a pure translation system. AB - Ribosome display is based on the concept that ternary complexes consisting of a nascent chain, ribosome, and mRNA can be generated, thereby establishing the linkage between genotype and phenotype that is essential for evolutionary experiments. With cell extract-based in vitro translation systems, it has been shown that ternary complexes can be generated by omitting the termination codon from the constructs, which can be stabilized at low temperature in the presence of high Mg2+ concentrations. Using an Escherichia coli-based reconstituted in vitro translation system (PURE system), in which all components necessary for the translation reaction were highly purified and reconstituted, ternary complexes could be generated equally well with a variety of sequences at the 3' end of the RNA, even those with a termination codon. Moreover, the generated complexes were stable at temperatures between 4 and 50 degrees C, and are thus highly stable in contrast to previous assumptions. PMID- 17134679 TI - Inhibition of the intestinal absorption of bile acids using cationic derivatives: mechanism and repercussions. AB - To pharmacologically interrupt bile acid enterohepatic circulation, two compounds named BAPA-3 and BAPA-6, with a steroid structure and 1 or 2 positive charges, were obtained by conjugation of N-(3-aminopropyl)-1,3-propanediamine with one or two moieties of glycocholic acid (GC). Both BAPA-3 and BAPA-6 inhibited Na+ dependent taurocholate (TC) uptake by Xenopus laevis oocytes expressing rat Asbt, with Ki values of 28 and 16 microM, respectively. BAPA-3 reduced Vmax without affecting Km. In contrast, BAPA-6 increased Km, with no effect on Vmax. Uptake of [14C]-GC by the last 10 cm of the rat ileum, perfused in situ over 60 min, was inhibited to a similar extent by unlabeled GC, BAPA-3 and BAPA-6. However, the intestinal absorption of these compounds was lower (BAPA-6) or much lower (BAPA 3) than that of GC. When administered orally to mice, both compounds (BAPA-3>BAPA 6) reduced the bile acid pool size, which was accompanied by up-regulation of hepatic Cyp7a1 and Hmgcr and intestinal Ostalpha/Ostbeta. A tendency towards a decreased expression of hepatic Ntcp and an enhanced expression of intestinal Asbt was also observed. Serum biochemical parameters were not affected by treatment with these compounds, except for a moderate increase in serum triglyceride concentrations. In sum, our results suggest that these compounds, in particular BAPA-3, are potentially useful tools for inhibiting the intestinal absorption of bile acids in a non-competitive manner. PMID- 17134680 TI - CagA protein of Helicobacter pylori: a hijacker of gastric epithelial cell signaling. AB - Epidemiological study has shown strong correlation between the Helicobacter pylori (H. pylori) infection and gastric carcinogenesis. However, the mechanism by which H. pylori induces gastric carcinogenesis is not known. In this review, we focused on the product of cytotoxin-associated gene A (CagA), one of the important virulence factors of H. pylori. H. pylori injects CagA protein into the host gastric epithelial cells through its needle-like structure, type IV secretion system. Injected CagA hijacks physiological signal transduction and causes pathological cellular response such as increased cell proliferation, motility, apoptosis and morphological change through different mechanisms. H. pylori has been shown to produce reactive oxygen species (ROS) in infected gastric mucosa. Although the main source of ROS production is possibly host neutrophil, we propose novel source of ROS production in this review; CagA itself can induce ROS production in gastric epithelial cell. Excessive ROS production in gastric epithelial cells can cause DNA damage and thus might involve in gastric carcinogenesis. Understanding the molecular mechanism by which H. pylori-induced carcinogenesis is important for developing new strategies against gastric cancer. PMID- 17134681 TI - Role of GABAergic antagonism in the neuroprotective effects of bilobalide. AB - Bilobalide, a constituent of Ginkgo biloba, has neuroprotective properties. Its mechanism of action is unknown but it was recently found to block GABA(A) receptors. The goal of this study was to test the potential role of a GABAergic mechanism for the neuroprotective activity of bilobalide. In rat hippocampal slices exposed to NMDA, release of choline indicates breakdown of membrane phospholipids. NMDA-induced choline release was almost completely blocked in the presence of bilobalide (10 microM) and under low-chloride conditions. Bicuculline (100 microM), a competitive antagonist at GABA(A) receptors, reduced NMDA-induced choline release to a small extent (-23%). GABA (100 microM) partially antagonized the inhibitory action of bilobalide. Exposure of hippocampal slices to NMDA also caused edema formation as measured by increases of tissue water content. NMDA induced edema formation was suppressed by bilobalide and by low-chloride conditions. Bicuculline exerted partial protection (by 30%) while GABA reduced bilobalide's effect by about one third. To investigate bilobalide's interaction with GABA(A) receptors directly, we measured binding of [(35)S]-TBPS to rat cortical membranes. TBPS binding was competitively inhibited by bilobalide in the low micromolar range (IC(50)=3.7 microM). As a functional test, we determined (36)chloride flux in rat corticohippocampal synaptoneurosomes. GABA (100 microM) significantly increased (36)chloride flux (+65%), and this increase was blocked by bilobalide, but with low potency (IC(50): 39 microM). We conclude that, while antagonism of GABA(A) receptors may contribute to bilobalide's neuroprotective effects, additional mechanisms must be postulated to fully explain bilobalide's actions. PMID- 17134682 TI - Corticospinal drive during painful voluntary contractions at constant force output. AB - In the voluntary contractions, output force can be maintained constant although the inhibitory influences exerted by pain on muscle activity. We investigated changes in the spontaneous and evoked activity of the abductor digiti minimi muscle (ADM) and the biceps brachii muscle (BIC) in healthy volunteers during constant force noxious contraction, resulting from chemically activated nociceptive afferents. EMG-force relationship, motor-evoked response (MEP) to transcranial magnetic stimulation and determinism (DET) of surface EMG signals during constant force contraction was analyzed before, during and after chemically induced tonic activation of their nociceptive afferents. Under constant force contraction, amplitude of surface EMG signal decreased in BIC and increased in ADM during pain with respect to control condition. In both muscles, the size of motor-evoked potential (MEP) induced by transcranial magnetic stimulation (TMS) of the primary motor cortex was significantly higher during pain than in control. Level of determinism extracted from surface EMG signal by non-linear method was similarly and significantly increased in both muscles during pain stimulation. Finally, nociceptive stimulation caused a decline in steadiness of the force exerted by ADM and BIC. These results are interpreted in terms of increased corticospinal synchronizing inputs. The possibility that it may play a role in governing force production to counteract pain inhibitory influences on motor system is considered. PMID- 17134683 TI - Ritter-based glycoconjugation of amino acids and peptides--access to novel glycoconjugates displaying a beta-amide linkage between amino acid and sugar moiety. AB - Beta-peptidic-D-gluco-, D-galacto-, and L-fuco-configured glycosyl amino acids can be prepared from the corresponding 2-deoxy-oct-3-ulopyranosonic acids via a one-pot intramolecular Ritter reaction. Initially, a ketopyranoside-based acid condenses under Lewis acid promoted conditions with nitriles (PhCN, MeCN) and a partially protected diamino ester (Boc-DAB-O-t-Bu, Boc-Orn-O-t-Bu) to form a beta peptidic glycosyl amino t-butylesters. The glycosyl amino t-butylesters can be converted into Fmoc-protected glycosyl amino acids that are suitably protected for solid-phase glycopeptide synthesis. Furthermore, replacement of the protected diamino ester by immobilized peptide amines permits post-synthetic N-terminal- and N(epsilon)-glycoconjugation of peptides on the solid phase. PMID- 17134684 TI - Structure of a highly pyruvylated galactan sulfate from the Pacific green alga Codium yezoense (Bryopsidales, Chlorophyta). AB - A polysaccharide fraction consisting of d-galactose, sulfate, and pyruvate in a molar proportion of 4:2:1 was isolated from the green seaweed Codium yezoense by water extraction followed by ion-exchange chromatography. To elucidate its structure, modified polysaccharides were prepared by desulfation, depyruvylation, and by total removal of non-carbohydrate substituents. Structures of the native polysaccharide and of the products of its chemical modifications were investigated by methylation analysis as well as by 1D and 2D (1)H and (13)C NMR spectroscopy. The polysaccharide devoid of sulfate and pyruvate was subjected to two subsequent Smith degradations to afford a rather low-molecular and essentially linear (1-->3)-beta-d-galactan. A highly ramified structure was suggested for the native polysaccharide, which contains linear backbone segments of 3-linked beta-d-galactopyranose residues connected by (1-->6) linkages, about 40% of 3-linked residues being additionally substituted at C-6, probably by short oligosaccharide residues also containing (1-->3) and (1-->6) linkages. Sulfate groups were found mainly at C-4 and in minor amounts at C-6. Pyruvate was found to form mainly five-membered cyclic ketals with O-3 and O-4 of the non-reducing terminal galactose residues. The minor part of pyruvate forms six-membered cyclic ketals with O-4 and O-6. The absolute configurations of ketals (R for six membered ketals and S for five-membered ones) were established using NMR spectral data. PMID- 17134685 TI - Myocardial angiogenesis after plasmid or adenoviral VEGF-A(165) gene transfer in rat myocardial infarction model. AB - OBJECTIVE: To compare gene transfer of plasmid (P) and adenovirus (Ad) encoding human vascular endothelial growth factor-A165 (hVEGF-A165) angiogenic efficacy and adverse effects as regards apoptosis and ectopic expression of the transgene in a rat myocardial infarction model. METHODS: Myocardial infarction was provoked in Fisher rats. One week later, PhVEGF-A165, PLacZ, AdhVEGF-A165, or AdLacZ was transferred intramyocardially along the border of the myocardial infarction. hVEGF-A expression was detected with ELISA. Myocardial vessel density was analyzed 1 and 4 weeks after gene transfer. Apoptosis was detected by TUNEL staining. Cardiac function was assessed with Tissue Doppler Velocity Imaging. RESULTS: Although AdhVEGF-A165 had substantially higher myocardial hVEGF-A expression than PhVEGF-A165, AdhVEGF-A165 and PhVEGF-A165 induced angiogenic effects to a similar extent with maintained increased arteriolar density after 4 weeks of gene transfer (p < 0.05). The two treatments also improved left ventricular function similarly. Adenoviral gene transfer induced a higher number of TUNEL positive cells than plasmid (p < 0.02). Ectopic expression of the transgene was present with both vectors but substantially higher after adenoviral gene transfer. CONCLUSIONS: AdhVEGF-A165 has no obvious angiogenic advantage over PhVEGF-A165 but more side effects at least in a rat myocardial infarction model. This indicates that PhVEGF-A165 might be more applicable for therapeutic angiogenesis than AdhVEGF-A165. PMID- 17134686 TI - Reduction of hyperacute rejection and protection of metabolism and function in hearts of human decay accelerating factor (hDAF)-expressing pigs. AB - OBJECTIVE: The use of pig hearts can solve the problem of shortage of donor hearts for transplantation. However, targeting rejection by single genetic modification was proven to be ineffective, highlighting the requirement for complex genetic modifications and more effective methods for transgenic animal production. We evaluated here whether hearts of hDAF transgenic pigs generated using our technique sperm-mediated gene transfer (SMGT) will be protected from structural damage, metabolic changes, and mechanical dysfunction during perfusion with human blood. METHODS: Hearts from control (C, n = 6) or transgenic (T, n = 5) pigs were perfused ex vivo for 4 h with fresh human blood using the ex vivo working mode system allowing monitoring of the function, metabolism, and structure. RESULTS: Cardiac output (mean+/-SEM) was maintained in T constant throughout the experiment, at 3.58+/-0.36 and 3.83+/-0.14 l/min after 30 min and 4 h, respectively, while cardiac output decreased to 1.95+/-0.35 l/min in C after 30 min of perfusion (p < 0.01 vs. T). The maximum increase in coronary perfusion pressure was reduced in T to 154+/-16% as compared to C (237+/-10%, p < 0.001). Myocardial ATP after 4 h was 21.1+/-1.1 nmol/mg dry wt (similar to initial) in T, while it decreased in C to 17.2+/-1.4 (p < 0.05). Deposition of complement factors C3 and C5b9 was present in C but not in T after perfusion. CONCLUSION: We have shown that hearts from hDAF transgenic pigs produced by SMGT are protected during perfusion with human blood and are metabolically stable and maintain mechanical function above the threshold level for life support. PMID- 17134687 TI - Shock-induced changes of Ca(i)2+ and Vm in myocyte cultures and computer model: Dependence on the timing of shock application. AB - OBJECTIVES: Responses of Ca(i)2+ to electrical shocks are believed to be important in defibrillation but measurements of shock-induced Ca(i)2+ changes during different phases of the action potential (AP) are lacking. The effects of shocks on Ca(i)2+ and Vm were investigated in geometrically defined cell cultures and in a computer model. METHODS: Uniform-field shocks (E = 10.4+/-0.9 V/cm) were applied 15-300 ms after AP upstroke in strands of cultured neonatal rat myocytes. Optical mapping was used to measure shock-induced Ca(i)2+ and Vm changes. A rat ionic model was used to elucidate ionic mechanisms of Ca(i)2+ responses. RESULTS: In experiments and simulations, shocks applied with short delays (15-40 ms) caused a transient decrease of Ca(i)2+ at sites of both DeltaV(+)m and DeltaV( )m. Simulations indicated that the Ca(i)2+ decrease at DeltaV(+)m sites was caused by reversed outward flow of L-type Ca2+ current (I(CaL)), while the Ca(i)2+ decrease at DeltaV(-)m sites was due to the NaCa exchanger (NCX). At intermediate delays (40-150 ms), shocks caused a Ca(i)2+ decrease at sites of DeltaV(-)m and an increase at sites of DeltaV(+)m. Simulations indicated that the Ca(i)2+ increase at DeltaV(+)m sites was caused by transient reactivation of I(CaL) combined with a reverse-mode operation of NCX. Shocks applied at long delays (150-300 ms) caused a Ca(i)2+ increase at DeltaV(+)m and no change at DeltaV(-)m sites. CONCLUSION: Effects of shocks on Ca(i)2+ depend on the timing of shock application. Shocks applied during the early AP cause a transient Ca(i)2+ decrease, while later in AP shocks induce a Ca(i)2+ increase at sites of DeltaV(+)m. Shock-induced Ca(i)2+ changes in different AP phases are primarily determined by combination of I(CaL) and NCX. PMID- 17134688 TI - Hepatocytes--the choice to investigate drug metabolism and toxicity in man: in vitro variability as a reflection of in vivo. AB - The pharmaceutical industry is committed to marketing safer drugs with fewer side effects, predictable pharmacokinetic properties and quantifiable drug-drug interactions. Drug metabolism is a major determinant of drug clearance and interindividual pharmacokinetic differences, and an indirect determinant of the clinical efficacy and toxicity of drugs. Progressive advances in the knowledge of metabolic routes and enzymes responsible for drug biotransformation have contributed to understanding the great metabolic variations existing in human beings. Phenotypic as well genotypic differences in the expression of the enzymes involved in drug metabolism are the main causes of this variability. However, only a minor part of phenotypic variability in man is attributable to gene polymorphisms, thus making the definition of a normal liver complex. At present, the use of human in vitro hepatic models at early preclinical stages means that the process of selecting drug candidates is becoming much more rational. Cultured human hepatocytes are considered to be the closest model to human liver. However, the fact that hepatocytes are located in a microenvironment that differs from that of the cell in the liver raises the question: to what extent does drug metabolism variability observed in vitro actually reflect that of the liver in vivo? By comparing the metabolism of a model compound both in vitro and in vivo in the same individual, a good correlation between the in vitro and in vivo relative abundance of oxidized metabolites and the hydrolysis of the compound was observed. Thus, it is reasonable to consider that the variability observed in human hepatocytes reflects the existing phenotypic heterogeneity of the P450 expression in human liver. PMID- 17134689 TI - Quantitative determination of gland mucous cells-type mucin using a monoclonal antibody, HIK1083: its pathophysiological changes in human gastric juice. AB - BACKGROUND: Pathological alteration in gastric mucosa is caused by Helicobacter pylori infection and is detectable by histological analysis. In particular, the alteration of gland mucous cells (GMCs)-type mucin, which plays a protective role against H. pylori infection, is critical in the pathogenesis of H. pylori-related gastritis. We established an assay for GMCs-type mucin and quantitatively assessed the pathophysiological changes in its content in human gastric juice samples. METHODS: The assay method for GMCs-type mucin was based on ELISA using a monoclonal antibody (HIK1083), and was used it to measure GMCs-type mucin in gastric juice obtained from patients with or without H. pylori infection. RESULTS: All the basic characteristics of the current method were satisfactory to quantify the GMCs-type mucin content in gastric juice. The GMCs-type mucin content, but not total mucin content, was significantly higher in patients with H. pylori infection (n=17; 437+/-476 U, mean+/-SD) than in those without H. pylori infection (n=55; 168+/-322 U, p<0.05). CONCLUSIONS: The current method is suitable for the quantitative analysis of GMCs-type mucin in gastric juice. The change in GMCs-type mucin content in gastric juice may be possibly implicated in the pathophysiology of the gastric mucosa and in the patient's gastric mucosal lesions. PMID- 17134690 TI - Fluorescent protein expression driven by her4 regulatory elements reveals the spatiotemporal pattern of Notch signaling in the nervous system of zebrafish embryos. AB - Notch activation inhibits neuronal differentiation during development of the nervous system; however, the dynamic role of Notch signaling in individual cell lineages remains poorly understood. We have characterized 3.4 kb 5'-regulatory sequence of a Notch target gene, her4, and used it to drive fluorescent gene expression in transgenic lines where the spatiotemporal pattern of Notch activation can be examined in vivo. The 3.4 kb her4 promoter contains five predicted Su(H) binding sites of which two proximal sites were confirmed to be required for Notch-mediated transcriptional activation. Without Notch, Su(H) effectively represses transcription regulated by the promoter. Analyses of transgenic zebrafish showed that while the expression of proneural genes and Notch activation were both critical for endogenous her4 expression, reporter gene expression was primarily regulated by Notch activity. This study also showed that her4 may be differently regulated in sensory cranial ganglia, where Notch activity is not essential for her4 expression and where Su(H) may repress her4 expression. The establishment of a reporter line with Notch-Su(H)-dependent fluorescent gene expression provides a tool to explore the complex role of Notch signaling in the development of vertebrate nervous system. PMID- 17134691 TI - Regulation of Xenopus gastrulation by ErbB signaling. AB - During Xenopus gastrulation, mesendodermal cells are internalized and display different movements. Head mesoderm migrates along the blastocoel roof, while trunk mesoderm undergoes convergent extension (C&E). Different signals are implicated in these processes. Our previous studies reveal that signals through ErbB receptor tyrosine kinases modulate Xenopus gastrulation, but the mechanisms employed are not understood. Here we report that ErbB signals control both C&E and head mesoderm migration. Inhibition of ErbB pathway blocks elongation of dorsal marginal zone explants and activin-treated animal caps without removing mesodermal gene expression. Bipolar cell shape and cell mixing in the dorsal region are impaired. Inhibition of ErbB signaling also interferes with migration of prechordal mesoderm on fibronectin. Cell-cell and cell-matrix interaction and cell spreading are reduced when ErbB signaling is blocked. Using antisense morpholino oligonucleotides, we show that ErbB4 is involved in Xenopus gastrulation morphogenesis, and it partially regulates cell movements through modulation of cell adhesion and membrane protrusions. Our results reveal for the first time that vertebrate ErbB signaling modulates gastrulation movements, thus providing a novel pathway, in addition to non-canonical Wnt and FGF signals, that controls gastrulation. We further demonstrate that regulation of cell adhesive properties and cell morphology may underlie the functions of ErbBs in gastrulation. PMID- 17134692 TI - The Drosophila casein kinase Iepsilon/delta Discs overgrown promotes cell survival via activation of DIAP1 expression. AB - The proper number of cells in developing tissues is achieved by coordinating cell division with apoptosis. In Drosophila, the adult wing is derived from wing imaginal discs, which undergo a period of growth and proliferation during larval stages without much programmed cell death. In this report, we demonstrate that the Drosophila casein kinase Iepsilon/delta, known as Discs overgrown (Dco), is required for maintaining this low level of apoptosis. Expression of dco can suppress the apoptotic activity of Head involution defective (Hid) in the developing eye. Loss of dco in the wing disc results in a dramatic reduction in expression of the caspase inhibitor DIAP1 and a concomitant activation of caspases. The regulation of DIAP1 by Dco occurs by a post-transcriptional mechanism that is independent of hid. Mutant clones of dco are considerably smaller than controls even when apoptosis is inhibited, suggesting that Dco promotes cell division/growth in addition to its role in cell survival. The dco phenotype cannot be explained by defects Wingless (Wg) signaling. We propose that Dco coordinates tissue size by stimulating cell division/growth and blocking apoptosis via activation of DIAP1 expression. PMID- 17134693 TI - Inhibitory effect of obovatol on nitric oxide production and activation of NF kappaB/MAP kinases in lipopolysaccharide-treated RAW 264.7cells. AB - The components of Magnolia obovata are known to have many pharmacological activities. In this study, we investigated the effects of obovatol, a neolignan compound isolated from the leaves of M. obovata, on nitric oxide (NO) production and NF-kappaB activity in lipopolysaccharide (LPS)-activated RAW 264.7 cells. The results show that obovatol (1-5 microM) significantly inhibited LPS-induced NO production in a concentration-dependent manner (IC(50): 0.91 microM). Consistent with the inhibitory effect on NO production, obovatol inhibits the expression of inducible nitric oxide synthase and cyclooxygenase-2 expression. Furthermore, obovatol suppressed NF-kappaB (p50 and p65) translocation to the nucleus as well as IkappaB release resulting in the inhibition of the DNA binding activity of the NF-kappaB. Obovatol also inhibited c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) signal, which are the most significantly involved signal in NO production and NF-kappaB activation. When the cells were treated with the combination of obovatol with U0126 (an ERK inhibitor) or SP600125 (a JNK inhibitor) as well as with SC-514 (an IKK2 inhibitor), much more inhibition of NO production was observed than that by obovatol alone. The present results suggest that obovatol has an inhibitory effect on NO production through the inhibition of NF-kappaB/MAPK activity, and thus can be used as an anti-inflammatory agent. PMID- 17134694 TI - The amiodarone derivative 2-methyl-3-(3,5-diiodo-4 carboxymethoxybenzyl)benzofuran (KB130015) opens large-conductance Ca2+-activated K+ channels and relaxes vascular smooth muscle. AB - 2-methyl-3-(3,5-diiodo-4-carboxymethoxybenzyl)benzofuran (KB130015) has been developed to retain the antiarrhythmic properties of the parent molecule amiodarone but to eliminate its undesired side effects. In patch-clamp experiments, KB130015 activated large-conductance, Ca2+-activated BK(Ca) channels formed by hSlo1 (alpha) subunits in HEK 293 cells. Channels were reversibly activated by shifting the open-probability/voltage (P(o)/V) relationship by about -60 mV in 3 muM intracellular free Ca2+ ([Ca2+]in). No effect on the single channel conductance was observed. KB130015-mediated activation of BK(Ca) channels was half-maximal at 20 microM with a Hill coefficient of 2.8. BK(Ca) activation by KB130015 did not require the presence of Ca2+ and still occurred with saturating (100 microM) [Ca2+]in. Effects of the prototypic BK(Ca) activator NS1619 (1,3-dihydro-1-[2-hydroxy-5-(trifluoromethyl)phenyl]-5-(trifluoromethyl) 2H-benzimidazol-2-one) and those of KB130015 were not additive suggesting that both activators may at least partially share a common mechanism of action. KB130015-mediated activation was observed also for BK(Ca) channels from insects and for human BK(Ca) channels with already profoundly left-shifted voltage dependence. In contrast, human intermediate conductance Ca2+-activated channels were inhibited by KB130015. Using segments of porcine pulmonary arteries, KB130015 induced endothelium-independent vasorelaxation, half-maximal at 43 microM KB130015. Relaxation was inhibited by 1 mM tetraethylammonium, suggesting that KB130015 can activate vascular smooth muscle type BK(Ca) channels under physiological conditions. Interestingly, the shift in the P(o)/V relationship was considerably stronger (-90 mV in 3 microM [Ca2+]in) for BK(Ca) channels containing Slo-beta1 subunits. Thus, KB130015 belongs to a novel class of BK(Ca) channel openers that exert an effect depending on the subunit composition of the channel complex. PMID- 17134695 TI - 11,13-dihydro-dehydroleucodine, a derivative of dehydroleucodine with an inactivated alkylating function conserves the anti-proliferative activity in G2 but does not cause cytotoxicity. AB - Modulation of vascular smooth muscle cell (VSMC) proliferation has critical therapeutic implications for vascular disease. Recently, we demonstrated that the sesquiterpene lactone dehydroleucodine (DhL) inhibited the proliferation of VSMCs in G2 phase. It is known that the alpha,beta-unsaturated carbonyl group of the sesquiterpene lactone has a nonspecific alkylating activity that inhibits a large number of enzymes or factors involved in key biological processes. We analyzed whether the DhL alpha-methylene-gamma-lactone function is directly involved in cell proliferation arrest in G2 and in cell toxicity. To this end, the effects of both DhL and 11,13-dihydro-dehydroleucodine (2H-DhL), a derivative of DhL with inactivated alpha-methylenelactone function, on cultured VSMC viability and proliferation were assessed. We found that both DhL and 2H-DhL inhibited the proliferation of VSMCs in a dose-dependent manner, inducing a transient arrest in G2 phase. DhL, but not 2H-DhL, had a cytotoxic effect at concentrations up to 12 microM, indicating that cell proliferation arrest and cytotoxicity are mediated by different cellular targets. From these results we infer that only 2H-DhL is able to arrest cell proliferation in G2 without affecting cell viability at any concentration. PMID- 17134696 TI - Bis-imide granulatimide analogues as potent Checkpoint 1 kinase inhibitors. AB - Granulatimide and isogranulatimide, natural products isolated from an ascidian, were found to be abrogators of the cell cycle G2-M phase checkpoint by inhibition of Checkpoint 1 kinase (Chk1). In the course of structure-activity relationship studies on granulatimide analogues, we have synthesized a series of bis-imides, in which the imidazole moiety was replaced by an imide heterocycle. Various modifications have been introduced on one or both imide heterocycles, on the benzene ring, and on the indole nitrogen. Moreover, aza bis-imide analogues were synthesized in which the indole moiety was replaced by a 7-azaindole. Compared to those of granulatimide and isogranulatimide, the Chk1 inhibitory activities of some of the bis-imide carbazoles were stronger. In particular, 1,3,4,6-tetrahydro 10-hydroxy-7H-dipyrrolo[3,4-a:3,4-c]carbazole-1,3,4,6-tetraone 11 exhibited an IC(50) value on purified full length Chk1 of 2 nM, which makes it a more potent Chk1 inhibitor than granulatimide and isogranulatimide. To get an insight into the selectivity of this new family of compounds, the inhibitory activities of 1,3,4,6-tetrahydro-7H-dipyrrolo[3,4-a:3,4-c]carbazole-1,3,4,6-tetraone A have been evaluated on a panel of 15 kinases, the strongest inhibitory potency was found for Chk1. The inhibitory activities of compounds A, 5 and 11 toward Src tyrosine kinase and the cytotoxicity of various tumor cell lines were also evaluated. PMID- 17134697 TI - Evidence of extra-telomeric effects of hTERT and its regulation involving a feedback loop. AB - The human telomerase reverse transcriptase (hTERT) is the catalytic subunit of the enzyme telomerase which is responsible for telomeric maintenance and extension. Using RNA interference to knock down hTERT mRNA expression, we provide evidence that hTERT exerts extra-telomeric effects on the cell cycle and on its own regulatory proteins, specifically: p53 and p21. We tested our hypothesis that hTERT regulates its own expression through effects on upstream regulatory genes using transformed human embryonic kidney (HEK 293) cells, p53 and p16(INK4a) null human ovarian cancer SKOV-3 cells, and p53-null MDA-MB-157 human mammary cancer cells. In HEK 293 cells, hTERT knockdown resulted in elevated p53 and p21 transcription and a decrease in cellular proliferation. Similar results were observed in the MDA-MB-157 cell line where p21 was upregulated, correlating with cell growth inhibition. In contrast, we observed a decrease in expression of p21 in SKOV-3 cells with hTERT knockdown and cell growth appeared to be unaffected. These findings suggest that hTERT may be involved in a feedback loop system, thereby playing a role in its own regulation. PMID- 17134698 TI - Reversible attenuation of neuropathic-like manifestations in rats by lesions or local blocks of the intralaminar or the medial thalamic nuclei. AB - BACKGROUND AND AIM: Thalamic somatosensory nuclei have been classified into medial and lateral systems based on their role in nociception. An imbalance between these two systems may result in abnormal somatic sensations and spontaneous pain. This study aims to investigate the effects of transient or permanent block of the medial and intralaminar nuclear groups on the neuropathic like behavior in a rat model for mononeuropathy. METHODS: Neuropathy was induced on one hind paw in different groups of rats following the spared nerve injury model. When the resulting hyperalgesia and allodynia (tactile and cold) reached a maximum plateau, the rats received either chemical or electrolytic lesion or lidocaine (2%) microperfusion, placed in the various thalamic nuclear groups. RESULTS: All procedures produced transient but significant decrease of neuropathic manifestations. The magnitude and duration of decrease depended on the type and the site of the block. These effects can be ranked in increasing order as follows, electrolyticor=1.3mm), 0.74 (0.38-1.42) by maximal common carotid artery IMT>or=75th percentile, and 1.09 (0.62-1.90) by extracranial carotid artery atherosclerotic score>or=2, with the use of multivariate logistic regression analyses. CONCLUSION: Hepatitis B virus seropositivity was not associated with an increased severity of carotid atherosclerosis in Taiwanese. PMID- 17134708 TI - Aggregated low density lipoproteins decrease metalloproteinase-9 expression and activity in human coronary smooth muscle cells. AB - Plaque stability largely depends on vascular smooth muscle cell (VSMC) function. VSMC secrete metalloproteinases (MMPs), matrix degrading endopeptidases, that regulate VSMC migration and function. Among them, gelatinase B or MMP-9 seems to have a protective effect by promoting a stable plaque phenotype. In macrophage foam cells oxidized LDL (oxLDL) uptake regulates MMP-9 expression. However, it is unknown whether VSMC-lipid loading by aggregated LDL (agLDL) internalization produces any effect on MMP-9 production by human resident vascular cells. In the present study, we analyzed the effect of lipid-internalization in MMP-9 and MMP-2 expression and activity and its consequences in VSMC migration. Our results show that agLDL-internalization down-regulates MMP-9 activity in a time-dependent manner up to 42% at 48h and in a dose-dependent manner up to 87% at 300 microg/mL. nLDL induced similar but not sustained decrease on MMP-9 activity. However, neither agLDL nor nLDL exerted any significant effect on MMP-2 and TIMP 1. VSMC regrowth after a scratch injury was significantly reduced by exposure to agLDL. We conclude that agLDL-lipid loading reduces MMP-9 activity and this effect is associated to inhibition of VSMC migration. Thus, agLDL internalization may have consequences on vascular remodeling after injury, and the stability of lipid-rich atherosclerotic plaques. PMID- 17134709 TI - Comparison of migration models for acidic solutes in micellar electrokinetic chromatography. AB - The validity of two models that explain the migration of ionisable solutes in micellar electrokinetic chromatography (MEKC), mobility model and retention factor model, has been tested. For this purpose, the mobility (mu) and retention factor (k) of a set of 10 phenolic compounds with different hydrophobicity and pKa values have been determined for several sodium dodecyl sulphate (SDS) concentrations and pH values, and fitted to the models. Results show that in general the retention factor model explains better the retention of ionisable solutes, although for hydrophilic compounds at low SDS concentration, mobility model can give better fits. The different drawbacks pointed out by several authors in relation to both models have been checked, and a deep evaluation of each one has been done. As a result we have observed that, while in the retention factor model the variation of k with pH and [SDS] always follows the same trend, the variation of mu with these variables mainly depends on the value of the binding constant of the neutral form of the solutes to the micelles, KHA(m), which plays a critical role in the fit of the mobility model. Also we provide rules and advices to set up the experimental conditions to apply each model to a particular solute. PMID- 17134710 TI - Direct determination of fourteen underivatized amino acids from Whitmania pigra by using liquid chromatography-evaporative light scattering detection. AB - A new reversed phase high performance liquid chromatography-evaporative light scattering detection (RP-HPLC-ELSD) method has been developed for the direct determination of fourteen amino acids in Whitmania pigra, a commonly used traditional Chinese medicine (TCM). Simultaneous separation of these fourteen amino acids was achieved on a Prevail C18 column. Under the condition of gradient elution, the fourteen amino acids were separated within 25 min. The drift tube temperature of ELSD was set at 115 degrees C, and with the nitrogen flow rate of 2.5 l/min. All calibration curves showed good linear regression (r > 0.9975) within test ranges. The recoveries ranged from 94.8% to 104.4%. The limit of detection (LOD) for each compound was more than 20 mg/l at a signal to noise ratio (S/N) of 3. The validated method was successfully applied to quantify fourteen amino acids from Whitmania pigra, which provides a new basis of overall assessment on quality of Whitmania pigra. PMID- 17134711 TI - Extraction of alkyl methylphosphonic acids from aqueous samples using a conventional polymeric solid-phase extraction sorbent and a molecularly imprinted polymer. AB - The analysis of alkyl methylphosphonic acids (AMPAs) constitutes an important subject for verifying the compliance to the Chemical Weapons Convention (CWC). Indeed, alkyl methylphosphonic acids are the degradation products of V and G nerve agents such as VX, sarin or soman. Lowering the limits of detection of analytical methods for complex aqueous matrices implies the introduction of concentration and clean-up steps in the whole analytical process. Therefore a molecular imprinted polymer (MIP) has been previously developed for the selective extraction and the concentration of the alkyl methylphosphonic acids. Unfortunately, the selective retention process on this MIP has involved the development of hydrogen bonds and so does not allow the direct percolation of aqueous samples. A change of solvent is then necessary and can be performed using solid-phase extraction (SPE) with conventional non selective hydrophobic sorbents. Two polymeric sorbents, Oasis HLB and HR-P resins, were selected for their high specific surface area. The extraction recoveries obtained on both sorbents were compared and the Oasis HLB sorbent was further selected and used for the percolation of acidified aqueous samples. An optimised SPE procedure was then applied to concentrate an aqueous soil extract spiked with isobutyl methylphosphonic acid (iBMPA) and cyclopentyl methylphosphonic acid (cPMPA) that was further cleaned-up by passing through the MIP. The resulting LC-MS full scan chromatograms highlight the clean-up effect of the SPE-MIP association by the removal of the matrix substances and the preservation of 95% of the compounds of interest. PMID- 17134712 TI - Fast and sensitive method to determine chloroanisoles in cork using an internally cooled solid-phase microextraction fiber. AB - A new generation of solid-phase microextraction (SPME) fiber, an internally cooled fiber (cold fiber with polydimethylsiloxane loading) that allows heating the sample matrix and simultaneously cooling the fiber coating, was used to determine 2,4-dichloroanisole, 2,6-dichloroanisole, 2,4,6-trichloroanisole and pentachloroanisole in cork. A comparison between the cold fiber and regular SPME fiber was performed. An automated headspace solid-phase microextraction (HS-SPME) using commercial fibers and an internally cooled SPME fiber (CF-HS-SPME) coupled to gas chromatography-time-of-flight mass spectrometry (GC-TOF-MS) was used. The extraction conditions for both CF-HS-SPME and HS-SPME were optimized using full factorial design and Doehlert matrix. The best extraction conditions for CF-HS SPME were obtained using 10 min of incubation time, 10 min of extraction time, and sample and fiber temperature of 130 and 10 degrees C, respectively. For HS SPME, polydimethylsiloxane/divinylbenzene (PDMS/DVB) fiber was used with 10 min of incubation time, 75 min of extraction time, 85 degrees C of sample temperature, 8 ml of water was added and agitated at 500 rpm. The quantification limits for the target compounds using CF-HS-SPME procedure were between 0.8 and 1.6 ng g(-1) of cork, while for HS-SPME were between 4 and 6 ng g(-1) of cork. Furthermore, the CF-HS-SPME procedure could be used as a non-destructive method after minor modification of the agitator for the autosampler. PMID- 17134713 TI - Characterization of reversible and pseudo-irreversible acetylcholinesterase inhibitors by means of an immobilized enzyme reactor. AB - The aim of the present study was the application of a human AChE-CIM-IMER (enzyme reactor containing acetylcholinesterase immobilized on a monolithic disk) for the rapid evaluation of the thermodynamic and kinetic constants, and the mechanism of action of new selected inhibitors. For this application, human recombinant AChE was covalently immobilized onto an ethylenediamine (EDA) monolithic Convective Interaction Media (CIM) disk and on-line studies were performed by inserting this IMER into a HPLC system. Short analysis time, absence of backpressure, low nonspecific matrix interactions and immediate recovery of enzyme activity were the best characteristics of this AChE-CIM-IMER. Mechanisms of action of selected reversible inhibitors (tacrine, donepezil, edrophonium, ambenonium) were evaluated by means of Lineweaver-Burk plot analysis. Analyses were performed on line by injecting increasing concentrations of the tested inhibitor and substrate and by monitoring the product peak area. AChE-CIM-IMER kinetic parameters (Km(app) and vmax(app)) were derived as well as inhibitory constants (Ki(app)) of selected compounds. Moreover, noteworthy results were obtained in the application of the AChE-CIM-IMER to the characterization of the carbamoylation and decarbamoylation steps in pseudo-irreversible binding of carbamate derivatives (physostigmine and rivastigmine). AChE-CIM-IMER appeared to be a valid tool to determine simultaneously the kinetic constants in a reliable and fast mode. The obtained values were found in agreement with those obtained with the classical methods with the free enzyme. Furthermore, after inactivation by carbamates, activity could be fully recovered and the AChE-CIM-IMER could be reused for further studies. Results showed that the AChE-CIM-IMER is a valid tool not only for automated fast screening in the first phase of the drug discovery process but also for the finest characterization of the mode of action of new hit compounds with increased accuracy and reproducibility and with saving of time and materials. PMID- 17134714 TI - Application of polymer based stationary phases in high performance liquid chromatography and capillary high performance liquid chromatography hyphenated to microcoil 1H nuclear magnetic resonance spectroscopy. AB - The increased demand for chromatographic materials that are able to achieve a fast separation of large quantities of structure analogues is a great challenge. It is known that polymer based chromatographic materials have a higher loadability, compared to silica based sorbents. Unfortunately these polymer materials cannot be used under high pressure which is necessary in order to obtain high flow rates, and hence long times are needed to perform a separation. However, by immobilizing a polymer on a mechanically stable porous silica core, this problem can be circumvented and higher flows become feasible on these materials. Especially for capillary liquid chromatography hyphenated with nuclear magnetic resonance a high loadability is of great importance in order to obtain sharp, resolved, and concentrated peaks thus resulting in a good signal to noise ratio in the NMR experiment. Therefore, a highly shape selective chromatographic sorbent was developed by covalently immobilizing a poly(ethylene-co-acrylic) acid copolymer (-CH(2)CH(2)-)(x)[CH(2)CH(CO(2)H)-](y) (x=119, y=2.4) with a mass fraction of acrylic acid of 5% as stationary phase on silica via a spacer molecule (3-glycidoxypropyltrimethoxysilane). First, the loadability of this sorbent compared to C(30) is demonstrated by the HPLC separation of two xanthophyll isomers. Subsequently, it has been successfully employed in the hyphenation of capillary HPLC with microcoil (1)H NMR spectroscopy by separating and identifying a highly concentrated solution of the tocopherol homologues. PMID- 17134715 TI - Development and implementation of a direct detection, quantitation and validation system for class I MHC self-peptide epitopes. AB - Gene and protein expression studies demonstrate that viral-infected and malignant cells undergo a complex series of transcriptional and translational changes. As class I MHC molecules reflect the proteome (and changes therein) by presenting intracellular peptide epitopes, the development of a direct discovery and validation technology for the identification of these epitopes is needed. We developed our technology using HIV-1-infected cells as a model. A combination of hollow fiber class I HLA protein production and mass spectrometric epitope analysis indicated a 3-fold increase in the host-peptide VLMTEDIKL(720-728), [eIF4G((720))] presented by the HLA-A*0201 of HIV-1-infected cells. This peptide is derived from the host-protein translation of eukaryotic initiation factor 4 gamma (eIF4G) that plays a pivotal role in cellular protein synthesis. Direct confirmation of expression of this self-encoded antigen was performed through development of a T cell receptor mimic (TCRm) monoclonal antibody (mAb). The resulting 4F7 TCRm demonstrated specific recognition of the eIF4G((720))-A*0201 complex. Staining of normal PBMCs with 4F7 showed only low levels of endogenous eIF4G((720)) presentation by HLA-A*0201, while 4F7 staining of HIV-1-infected PBMCs revealed an approximately 3-fold increase in eIF4G((720))-A*0201. The MHC peptide complex was initially detectable by 4F7 at 3 days post-infection, with a steady increase through day 8. We therefore demonstrate the successful development and implementation of an integrated discovery and validation technology system for direct identification and confirmation of class I MHC peptide epitopes on cells. PMID- 17134716 TI - Lysozyme amyloid oligomers and fibrils induce cellular death via different apoptotic/necrotic pathways. AB - Among the newly discovered amyloid properties, its cytotoxicity plays a key role. Lysozyme is a ubiquitous protein involved in systemic amyloidoses in vivo and forming amyloid under destabilising conditions in vitro. We characterized both oligomers and fibrils of hen lysozyme by atomic force microscopy and demonstrated their dose (5-50 microM) and time-dependent (6-48 h) effect on neuroblastoma SH SY5Y cell viability. We revealed that fibrils induce a decrease of cell viability after 6 h due to membrane damage shown by inhibition of WST-1 reduction, early lactate dehydrogenase release, and propidium iodide intake; by contrast, oligomers activate caspases after 6 h but cause the cell viability to decline only after 48 h, as shown by fluorescent-labelled annexin V binding to externalized phosphatidylserine, propidium iodide DNA staining, lactate dehydrogenase release, and by typical apoptotic shrinking of cells. We conclude that oligomers induce apoptosis-like cell death, while the fibrils lead to necrosis-like death. As polymorphism is a common property of an amyloid, we demonstrated that it is not a single uniform species but rather a continuum of cross-beta-sheet-containing amyloids that are cytotoxic. An abundance of lysozyme highlights a universal feature of this phenomenon, indicating that amyloid toxicity should be assessed in all clinical applications involving proteinaceous materials. PMID- 17134717 TI - Molecular mechanism of the regulation of Bacillus subtilis gltAB expression by GltC. AB - In Bacillus subilis, glutamate synthase, a major enzyme of nitrogen metabolism, is encoded by the gltAB operon. Significant expression of this operon requires the activity of GltC, a LysR-family protein, encoded by the divergently transcribed gene. We purified a soluble, active form of GltC and found that it requires alpha-ketoglutarate, a substrate of glutamate synthase, to fully activate gltA transcription in vitro, and that its activity is inhibited by glutamate, the product of glutamate synthase. GltC regulated gltAB transcription through binding to three dyad-symmetry elements, Box I, Box II and Box III, located in the intergenic region of gltC and gltA. Free GltC bound almost exclusively to Box I and activated gltAB transcription only marginally. Glutamate bound GltC bound to Box I and Box III, and repressed gltAB transcription. In the presence of alpha-ketoglutarate, GltC bound to Box I and Box II, stabilized binding of RNA polymerase to the gltA promoter, and activated gltAB transcription. The binding of GltC to Box II, which partially overlaps the -35 region of the gltA promoter, seems to be essential for activation of the gltAB operon. Due to the high concentration of glutamate in B. subtilis cells grown under most conditions, alterations of the concentration of alpha-ketoglutarate seem to be crucial for modulation of GltC activity and gltAB expression. PMID- 17134718 TI - Actin filament severing by cofilin. AB - Cofilin is essential for cell viability and for actin-based motility. Cofilin severs actin filaments, which enhances the dynamics of filament assembly. We investigated the mechanism of filament severing by cofilin with direct fluorescence microscopy observation of single actin filaments in real time. In cells, actin filaments are likely to be attached at multiple points along their length, and we found that attaching filaments in such a manner greatly increased the efficiency of filament severing by cofilin. Cofilin severing increased and then decreased with increasing concentration of cofilin. Together, these results indicate that cofilin severs the actin filament by a mechanism of allosteric and cooperative destabilization. Severing is more efficient when relaxation of this cofilin-induced instability of the actin filament is inhibited by restricting the flexibility of the filament. These conclusions have particular relevance to cofilin function during actin-based motility in cells and in synthetic systems. PMID- 17134719 TI - Self-masking in an intact ERM-merlin protein: an active role for the central alpha-helical domain. AB - Ezrin/radixin/moesin (ERM) family members provide a regulated link between the cortical actin cytoskeleton and the plasma membrane to govern membrane structure and organization. Here, we report the crystal structure of intact insect moesin, revealing that its essential yet previously uncharacterized alpha-helical domain forms extensive interactions with conserved surfaces of the band four-point one/ezrin/radixin/moesin (FERM) domain. These interdomain contacts provide a functional explanation for how PIP(2) binding and tyrosine phosphorylation of ezrin lead to activation, and provide an understanding of previously enigmatic loss-of-function missense mutations in the tumor suppressor merlin. Sequence conservation and biochemical results indicate that this structure represents a complete model for the closed state of all ERM-merlin proteins, wherein the central alpha-helical domain is an active participant in an extensive set of inhibitory interactions that can be unmasked, in a rheostat-like manner, by coincident regulatory factors that help determine cell polarity and membrane structure. PMID- 17134720 TI - Constitutive promoter occupancy by the MBF-1 activator and chromatin modification of the developmental regulated sea urchin alpha-H2A histone gene. AB - The tandemly repeated sea urchin alpha-histone genes are developmentally regulated. These genes are transcribed up to the early blastula stage and permanently silenced as the embryos approach gastrulation. As previously described, expression of the alpha-H2A gene depends on the binding of the MBF-1 activator to the 5' enhancer, while down-regulation relies on the functional interaction between the 3' sns 5 insulator and the GA repeats located upstream of the enhancer. As persistent MBF-1 binding and enhancer activity are detected in gastrula embryos, we have studied the molecular mechanisms that prevent the bound MBF-1 from trans-activating the H2A promoter at this stage of development. Here we used chromatin immunoprecipitation to demonstrate that MBF-1 occupies its site regardless of the transcriptional state of the H2A gene. In addition, we have mapped two nucleosomes specifically positioned on the enhancer and promoter regions of the repressed H2A gene. Interestingly, insertion of a 26 bp oligonucleotide between the enhancer and the TATA box, led to up-regulation of the H2A gene at gastrula stage, possibly by changing the position of the TATA nucleosome. Finally, we found association of histone de-acetylase and de acetylation and methylation of K9 of histone H3 on the promoter and insulator of the repressed H2A chromatin. These data argue for a role of a defined positioned nucleosome in the promoter and histone tail post-translational modifications, in the 3' insulator and 5' regulatory regions, in the repression of the alpha-H2A gene despite the presence of the MBF-1 activator bound to the enhancer. PMID- 17134721 TI - Sec2 is a highly efficient exchange factor for the Rab protein Sec4. AB - Sec2 is a reversibly membrane associated multi-domain protein with guanine nucleotide exchange activity towards the yeast Rab-protein Sec4. Both proteins are localized to secretory vesicles destined for exocytosis. We have used transient kinetic methods to show that Sec2 is a highly active exchange factor, in contrast to other proteins previously characterized as Rab exchange factors. With a K(d) value for the Sec2:Sec4.GDP interaction of ca 70 microM and a maximal rate of GDP displacement of ca 15 s(-1), it is 100-1000-fold more effective than other proteins showing exchange activity towards Rabs (MSS4, DSS4, Vps9) and ca tenfold faster than Cdc25 as a Ras specific exchanger, although still 100-fold slower than the fastest systems studied so far, EF-Tu/Ef-Ts and Ran/RCC1. A comparison with other proteins showing Rab exchange activity shows that maximal rates of GDP dissociation catalyzed by Sec2 are orders of magnitude faster. When comparing Sec2 with DSS4, which also acts on Sec4, the difference was particularly dramatic. Another difference is seen in the kinetics of association of GTP with the Sec4:Sec2 complex, a process which is extremely slow for DSS4/MSS4 complexes with cognate Rabs but in the range observed for other GTPase:exchanger complexes for Sec4:Sec2., It is suggested that systems such as Ef-Tu/Ef-Ts and Ran/RCC1 have evolved for maximal possible activity for the interaction between two soluble proteins, whereas other evolutionary constraints which are connected to the spatial and temporal coordination of events in vesicular transport and other regulatory networks have determined the detailed kinetic properties of the other systems. PMID- 17134722 TI - Beta-blockers reduce the risk of early death in ischemic stroke. AB - OBJECTIVES: Beta-blockers reduce mortality in patients after myocardial infarction. Experimental studies suggest that beta-blockers have also neuroprotective properties. The aim of this study was to assess if use of beta blockers is associated with reduced risk of early death in ischemic stroke patients. MATERIALS AND METHODS: Retrospective data analysis of 841 consecutive patients with acute ischemic stroke admitted to the stroke unit within 24 h after stroke onset. RESULTS: 10.6% of patients received beta-blockers during hospitalization. Thirty-day case fatality was significantly lower in patients treated with beta-blockers than in those not treated with beta-blockers (6.8% versus 19.0%, P < 0.01). After adjustment for other prognostic factors, the use of beta-blockers was associated with reduced risk of early death (relative hazard 0.37, 95% confidence interval 0.16-0.84) independently of age, stroke severity, fasting glucose, total cholesterol level and pneumonia. When patients who died of cardiovascular causes were excluded from the analysis, the use of beta-blocker was no longer significantly associated with risk of death (P = 0.12). CONCLUSION: In a retrospective series the use of beta-blockers was associated with reduced risk of early death in patients with ischemic stroke. PMID- 17134723 TI - Bilateral and unilateral movement training on upper limb function in chronic stroke patients: A TMS study. AB - The use of activity-dependent interventions has shown some success in promoting recovery of upper limb function in chronic stroke patients. This study compared the neurophysiological and behavioural changes associated with two such rehabilitation protocols: unilateral and bilateral movement training. Twelve chronic stroke patients were randomly assigned to the two training protocols involving six daily practice sessions. Each session consisted of 50 trials of a dowel placement task performed either with both impaired and unimpaired arm moving synchronously (bilateral training group) or with only the impaired arm moving (unilateral training). Kinematic measurements of upper limb movements were made in four unilateral test trials performed prior to and following each practice session. Functional assessments of the impaired upper limb and neurophysiological assessments, using transcranial magnetic stimulation (TMS), of the affected and non-affected cortical hemispheres were made prior to and following the intervention sessions. Individuals receiving bilateral training showed a reduction in movement time of the impaired limb and increased upper limb functional ability compared to individuals receiving unilateral training. In some patients changes to upper limb function were associated with changes to the cortical representation of a target muscle in the non-affected hemisphere. Overall, these findings suggest that a short-term bilateral training intervention may be effective in facilitating upper limb motor function in chronic stroke patients. PMID- 17134724 TI - A study of CSF catecholamine and its metabolites in acute and convalescent period of encephalitis. AB - AIMS: To evaluate cerebrospinal fluid (CSF) catecholamine (CA) and its metabolites in encephalitis patients in acute and convalescent period and correlate these with clinical and magnetic resonance imaging (MRI) features. SUBJECTS AND METHODS: Patients with acute encephalitis diagnosed on the basis of clinical, CSF, MRI and virological parameters underwent detailed neurological evaluation including Glasgow Coma Scale (GCS), Unified Parkinson's Disease Rating Scale (UPDRS) and Dystonia Rating Scale. Cranial MRI was carried out and CSF dopamine (DA), norepinephrine (NE), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 5-hydroxytryptamine (5HT) levels were estimated by High Performance Liquid Chromatography (HPLC). The CSF catecholamine levels were compared with convalescent phase as well as with controls. These levels were also correlated with parkinsonian features, dystonia and radiological abnormalities. RESULTS: There were 29 encephalitis patients; whose age ranged between 2 and 65 years, 4 were females and 11 children. 25 patients had Japanese encephalitis (JE) and 4 nonspecific encephalitis. The mean GCS score was 8 and 13 had seizures. Movement disorders were present in 13 patients and included parkinsonian features in 5, dystonia in 1 and combination of both in 7 patients. MRI revealed abnormalities in 15 out of 21 patients and included thalamic lesion in 10, globus pallidus in 4, putamen in 5, caudate in 4 and midbrain in 9 patients. In acute stage NE, DOPAC, 5HT and HVA levels were significantly lower compared to controls. NE levels significantly correlated with dystonia and thalamic lesions. Convalescent CSF study revealed significantly lower levels of DOPAC compared to acute phase. CSF catecholamine levels in encephalitis patients with and without movement disorders were not significantly different. CONCLUSION: In encephalitis, catecholamine and its metabolites are lower in acute and convalescent phase. Norepinephrine level correlates with dystonia and thalamic lesions. PMID- 17134725 TI - Ischemia/reperfusion in rat heart induces leptin and leptin receptor gene expression. AB - Concentrations of leptin, an adipocyte-derived hormone, are elevated in obesity. Recently, leptin has been shown to participate in multiple biological actions including inflammation, reproduction, and angiogenesis. Leptin has also been documented as a critical component in the process of wound healing; however, leptin involvement in cardiovascular disease is poorly understood. We examined the expression of leptin (ob) and leptin receptor (ob-R) genes in the rat heart following ischemia/reperfusion, which was induced by coronary artery ligation, and mRNA was obtained from hearts 0.5 to 36 h after initiating reperfusion. Expressions of ob and ob-R mRNA were examined by real-time quantitative RT-PCR and immunohistochemistry. The ob and ob-Ra mRNA and protein expressions were significantly increased (p<0.01) and ob-Rb mRNA was significantly decreased (p<0.01) in hearts after 8 h of reperfusion. Furthermore, ob and ob-R proteins were expressed in injured myocytes where inflammatory cells infiltrated. In contrast, those expressions were not influenced in hearts after 8 h of ischemia stress only. To determine the functional effects of leptin on the ischemic/reperfused heart, rats were treated with anti-leptin antibodies prior to ischemia/reperfusion; however, this treatment did not affect the elevation of mRNA expression levels of inflammatory markers such as TNF-alpha and IL-1beta in ischemic hearts. Our results demonstrated for the first time that ischemia/reperfusion induced leptin and leptin receptor gene expression in the rat heart. This study helps to elucidate the mechanisms behind the onset and development of ischemic heart disease concomitant with obesity. PMID- 17134726 TI - Non-targeted bystander effects induced by ionizing radiation. AB - Radiation-induced bystander effects refer to those responses occurring in cells that were not subject to energy deposition events following ionizing radiation. These bystander cells may have been neighbors of irradiated cells, or physically separated but subject to soluble secreted signals from irradiated cells. Bystander effects have been observed in vitro and in vivo and for various radiation qualities. In tribute to an old friend and colleague, Anthony V. Carrano, who would have said "well what are the critical questions that should be addressed, and so what?", we review the evidence for non-targeted radiation induced bystander effects with emphasis on prevailing questions in this rapidly developing research field, and the potential significance of bystander effects in evaluating the detrimental health effects of radiation exposure. PMID- 17134727 TI - Trivalent arsenicals induce lipid peroxidation, protein carbonylation, and oxidative DNA damage in human urothelial cells. AB - Drinking arsenic-contaminated water is associated with an increased risk of bladder cancer. Arsenate (iAs(V)), arsenite (iAs(III)), monomethylarsonous acid (MMA(III)), monomethylarsonic acid (MMA(V)), dimethylarsinous acid (DMA(III)), and dimethylarsinic acid (DMA(V)) have all been detected in the urine of people who drink arsenic-contaminated water. The aim of this research was to investigate which of these arsenicals are more hazardous to human urothelial cells. The results indicate that iAs(III), MMA(III), and DMA(III) were more potent in inducing cytotoxicity, lipid peroxidation, protein carbonylation, oxidative DNA damage, nitric oxide, superoxide, hydrogen peroxide, and cellular free iron than MMA(V), DMA(V), and iAs(V) in human urothelial carcinoma and transformed cells. However, the results did not show convincingly that the trivalent arsenicals were more potent than pentavalent arsenicals in decreasing the intracellular contents of total thiol, protein thiol, and reduced glutathione. Induction of oxidative DNA damage was observed with 0.2 microM of iAs(III), MMA(III), or DMA(III) as early as 1h. Because of its high oxidative damage, higher proportion in urine, and lower cytotoxicity, DMA(III) may be the most hazardous arsenical to human urothelial cells. PMID- 17134728 TI - Multimammate rat (Mastomys natalensis), Tristram's jird (Meriones tristrami) and Wagner's gerbil (Gerbillus dasyurus) as laboratory models of acute neosporosis. AB - To test the different sensitivity of rodents of the subfamily Murinae and Gerbillinae, Wagner's gerbils (Gerbillus dasyurus), Tristram's jirds (Meriones tristrami) and multimammate rats (Mastomys natalensis) were inoculated with Neospora caninum tachyzoites. Clinical signs of neosporosis appeared in all inoculated animals. Histopathological examination confirmed the presence of tachyzoites in brains, lungs, skeletal muscle, myocardium, liver, in serosa of stomach and intestines, and in vesicular accessory genital glands. An examination of brains by PCR revealed presence of N. caninum DNA in all experimentally N. caninum infected rodents. The susceptibility of Wagner's gerbils and Tristram's jirds further proved the high sensitivity of gerbiline rodents to the N. caninum infection. The finding of N. caninum tachyzoites in the vesicular accessory genital glands of the infected gerbils suggests the usefulness of the rodent model for demonstration of N. caninum in the male reproductive system. Moreover, the multimammate rat was a susceptible experimental host to be the first immunocompetent rodent of the subfamily Murinae. PMID- 17134729 TI - Complement inactivating proteins and intraspecies venom variation in Crotalus oreganus helleri. AB - Complement inactivating properties were detected in venom from the southern California distribution of Crotalus oreganus helleri (Southern Pacific Rattlesnake). This activity showed strong geographic bias to the San Bernardino Mountain range, and venom from this area reacted strongly with Fraction 5 antiserum (AF5). However, venoms from the San Jacinto Mountain range, which have been previously shown to contain Mojave toxin, did not inhibit complement and did not react with AF5. AF5 has been previously shown to recognize a protease in C. scutulatus venom that inactivates complement, but the identity of this protein has remained unknown. Using a functional venomic approach, utilizing two dimensional gel electrophoresis coupled with liquid chromatography and tandem mass spectrometry (LC/MS/MS), we have identified catrocollastatin and hemorrhagic toxin II (HT-2) as the primary proteins recognized by AF5. The information we present within this manuscript further illustrates the now well-known reality of intraspecies venom variation and the challenges faced in providing comprehensive polyvalent antivenoms. PMID- 17134730 TI - Palmitoylation of the cysteine-rich endodomain of the SARS-coronavirus spike glycoprotein is important for spike-mediated cell fusion. AB - The SARS-coronavirus (SARS-CoV) is the etiological agent of the severe acute respiratory syndrome (SARS). The SARS-CoV spike (S) glycoprotein mediates membrane fusion events during virus entry and virus-induced cell-to-cell fusion. The cytoplasmic portion of the S glycoprotein contains four cysteine-rich amino acid clusters. Individual cysteine clusters were altered via cysteine-to-alanine amino acid replacement and the modified S glycoproteins were tested for their transport to cell-surfaces and ability to cause cell fusion in transient transfection assays. Mutagenesis of the cysteine cluster I, located immediately proximal to the predicted transmembrane, domain did not appreciably reduce cell surface expression, although S-mediated cell fusion was reduced by more than 50% in comparison to the wild-type S. Similarly, mutagenesis of the cysteine cluster II located adjacent to cluster I reduced S-mediated cell fusion by more than 60% compared to the wild-type S, while cell-surface expression was reduced by less than 20%. Mutagenesis of cysteine clusters III and IV did not appreciably affect S cell-surface expression or S-mediated cell fusion. The wild-type S was palmitoylated as evidenced by the efficient incorporation of (3)H-palmitic acid in wild-type S molecules. S glycoprotein palmitoylation was significantly reduced for mutant glycoproteins having cluster I and II cysteine changes, but was largely unaffected for cysteine cluster III and IV mutants. These results show that the S cytoplasmic domain is palmitoylated and that palmitoylation of the membrane proximal cysteine clusters I and II may be important for S-mediated cell fusion. PMID- 17134731 TI - The West Nile virus mutant spectrum is host-dependant and a determinant of mortality in mice. AB - To define the impact of mosquitoes and birds on intrahost WNV population dynamics, the mutant spectra that arose as a result of 20 serial in vivo passages in Culex pipiens and young chickens were examined. Genetically homogeneous WNV was serially passaged 20 times in each host. Genetic diversity was greater in mosquito-passaged WNV compared to chicken-passaged WNV. Changes in the viral consensus sequence occurred in WNV passaged in mosquitoes earlier and more frequently than in chicken-passaged WNV. Analysis of synonymous and nonsynonymous variation suggested that purifying selection was relaxed during passage in mosquitoes. Mortality in mice was significantly negatively correlated with the size of the WNV mutant spectrum. These studies suggest that mosquitoes serve as sources for WNV genetic diversity, that birds are selective sieves, and that both the consensus sequence and the mutant spectrum contribute to WNV phenotype. PMID- 17134732 TI - PCB decomposition and formation in thermal treatment plant equipment. AB - In this study we investigated both the decomposition and unintentional formation of polychlorinated biphenyl congeners during combustion experiments of refuse derived fuel (RDF) and automobile shredder residue (ASR) at several stages in thermal treatment plant equipment composed of a primary combustion chamber, a secondary combustion chamber, and other equipments for flue gas treatment. In both experiments, the unintentional formation of PCB occurred in the primary combustion chamber at the same time as the decomposition of PCB in input samples. By combusting RDF, non-ortho-PCB predominantly formed, whereas ortho-PCB and symmetric chlorinated biphenyls (e.g., #52/69, #87/108, and #151) tended to be decomposed. ASR formed the higher chlorinated biphenyls more than RDF. These by products from ASR had no structural relation with ortho-chlorine. Lower chlorinated biphenyls appeared as predominant homologues at the final exit site, while all congeners from lower to higher chlorinated PCB were unintentionally formed as by-products in the primary combustion chamber. This result showed that the flue gas treatment equipments effectively removed higher chlorinated PCB. Input marker congeners of RDF were #11, #39, and #68, while those for ASR were #11, #101, #110/120, and #118. Otherwise, combustion marker congeners of RDF were #13/12, #35, #77, and #126, while those for ASR were #170, #194, #206, and #209. While the concentration of PCB increased significantly in the primary combustion chamber, the value of toxicity equivalency quantity for dioxin-like PCB decreased in the secondary combustion chamber and the flue gas treatment equipments. PMID- 17134733 TI - Diagnosis of ectopic tissue versus contamination by genetic fingerprinting in a routine surgical pathology specimen. AB - A 62-year-old woman presented with recurring right upper quadrant pain and underwent a routine laparoscopic cholecystectomy. A large gallstone was found impacted in the fundus of the gall bladder. Interestingly, aside from noting mild inflammation of the gall bladder wall, microscopic examination of the specimen identified 2 fragments of benign thyroid tissue. Given the routine nature of the surgical procedure and lack of abnormality detected during the operation, the attending pathologist suspected extraneous tissue contamination ("floater") of the pathology specimen and submitted the block and slides to Molecular Pathology. The thyroid tissue-containing fragments and gallbladder wall were independently isolated and subjected to genetic fingerprinting using a standard forensic DNA identification panel. All fragments showed the identical fingerprint, strongly suggesting that they belonged to the same patient. The results indicated that the thyroid tissue was from an ectopic rest adjacent to the gall bladder, which has been reported only very rarely in the previous literature and illustrates the unusual use of molecular genetic testing to confirm the presence of ectopic tissue versus contamination. PMID- 17134734 TI - Cytogenetic analysis of B-cell posttransplant lymphoproliferations validates the World Health Organization classification and suggests inclusion of florid follicular hyperplasia as a precursor lesion. AB - Cytogenetic abnormalities in B-cell posttransplant lymphoproliferative disorders (PTLD) have not been well characterized. We thus performed cytogenetic analysis of 28 cases of B-cell PTLD, 1 infectious mononucleosis (IM)-like lesion, 9 polymorphic PTLD, 17 monomorphic PTLD, and 1 classical Hodgkin lymphoma (HL), and correlated the karyotypic findings with the phenotype, Epstein-Barr virus infection status, and clinical outcome. Karyotypes of 19 cases of posttransplant florid follicular hyperplasia (FFH) were also analyzed. Informative karyotypes were obtained in 20 (71.4%) of 28 PTLDs and 18 (94.7%) of 19 FFHs. Clonal karyotypic abnormalities were detected in 13 (65%) of 20 PTLDs, including 9 (75%) of 12 monomorphic PTLDs, 2 (33.3%) of 6 polymorphic PTLDs, 1 IM-like lesion, and 1 HL, and 2 (11.1%) of 18 FFHs. Recurrent chromosome breaks at 1q11-21 (n = 6, including 1 FFH), 14q32 (n = 3, including 1 FFH), 16p13 (n = 3), 11q23-24 (n = 2), and 8q24 (c-MYC) (n = 2); gains of chromosome 7 (n = 4), X (n = 3), 2 (n = 3), 12 (n = 2); and loss of chromosome 22 (n = 2, including 1 IM-like lesion) were identified. The presence of cytogenetic abnormalities did not correlate with PTLD phenotype, Epstein-Barr virus infection, or clinical outcome. We describe novel karyotypic aberrations in PTLD and report clonal cytogenetic abnormalities in posttransplant FFH and an IM-like lesion for the first time. Our findings provide validation of the current World Health Organization classification of PTLD and also suggest incorporation of FFH as the earliest recognizable precursor of PTLD. PMID- 17134735 TI - Interphase fluorescence in situ hybridization is more sensitive than BIOMED-2 polymerase chain reaction protocol in detecting IGH-BCL2 rearrangement in both fixed and frozen lymph node with follicular lymphoma. AB - The detection of t(14;18)(q32;q21) is advisable for the diagnosis of follicular lymphoma (FL). In 51 patients with FL, we evaluated the applicability and sensitivity of interphase fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) using commercially available reagents. In 23 patients, only a formalin-fixed lymph node was available. In 28 patients, both frozen and formalin-fixed lymph nodes were evaluated. Fluorescence in situ hybridization was found to be 100% applicable whatever the material type. With the use of IGH-BCL2 dual-fusion, dual-color probes, t(14;18) translocation was detected in 47 (92%) of 51 FL cases with concordant results between isolated nuclei (n = 41) and frozen cytologic imprints (n = 28). Twenty-two IGH-BCL2 positive cases were also studied on fixed sections with BCL2 split signal probes showing a BCL2 split in all. Conversely, no BCL2 split was observed in IGH-BCL2 negative cases (n = 4). Owing to DNA degradation as assessed by the failure of control genes amplification, the applicability of PCR was found to be 76% in fixed lymph nodes (n = 51). After exclusion of the 12 noninformative cases, the BIOMED-2 protocol allowed the detection of an IGH-BCL2 fusion in 25 (64%) of 39 fixed specimens with 11 PCR-negative (31%) of 36 FISH-positive cases. Even on frozen material with 100% applicability, the amplification of a BCL2-JH breakpoint was achieved in only 20 (71%) of 28 cases with 5 PCR-negative (20%) out of 25 FISH-positive cases. Therefore, FISH was found superior to PCR (using BIOMED-2 protocol) in detecting IGH-BCL2 fusion. Finally, FISH individualized 4 IGH-BCL2-negative FL cases without specific histopathologic features. With the use of split signal DNA probes, 1 case showed a trisomy of the BCL2 locus and another displayed BCL6 and IGH breakpoints that would suggest a t(3;14). Whether such IGH-BCL2-negative cases are characterized by alternative oncogenetic pathways remains to be determined. PMID- 17134736 TI - Alpha-methylacyl-CoA racemase (P504S)/34betaE12/p63 triple cocktail stain in prostatic adenocarcinoma after hormonal therapy. AB - Alpha-methylacyl-CoA racemase (AMACR) has recently been shown to be a highly sensitive marker for the diagnosis of prostate cancer. However, there is limited information concerning its utility as a marker for prostate carcinoma after hormonal therapy. Our current investigation was conducted to evaluate the expression of AMACR in patients with prostate carcinoma after hormonal therapy and assess its diagnostic utility in combination with p63 and high molecular weight cytokeratin (34betaE12) staining. Prostate tissues from 49 patients who had been treated with hormonal therapy were immunohistochemically analyzed for AMACR, 34betaE12, and p63 expression by a triple antibody cocktail stain. The staining intensities and the percentages of positively staining tumor cells were recorded. The correlations between AMACR expression and metastatic status, associated hormonal therapy regimens, and the extent of hormone therapy effect were analyzed. All malignant acini were completely negative for both basal cell markers (34betaE12 and p63). Tumor cells failed to demonstrate expression of AMACR in 14 (29%) of 49 cases. In the remaining 35 cases (71%), positive immunostaining for AMACR was noted, but with variable intensities and percentages of cells stained. Positive staining for AMACR in benign glands was not seen in any case. In all cases, basal cells were strongly stained by p63 in benign acini with a mean positive percentage of 96%. Similarly, basal cells in benign acini displayed moderate staining intensities for 34betaE12 in 3 (7%) of 41 cases and strong immunostaining for this marker in the remaining 38 cases (93%); the mean percentage of positive cells was 92%. alpha-methylacyl-CoA racemase expression may be substantially diminished or entirely lost in prostate carcinoma after hormonal therapy. This variation in AMACR expression does not correlate with the metastatic status, the modality of hormonal therapy, or the extent of therapy related effect. It is important that pathologists be aware that some hormonally treated prostate carcinomas do not express AMACR, and that immunostaining in such cases must be interpreted with caution. A triple cocktail stain using AMACR, 34betaE12, and p63 can be helpful in evaluating prostate specimens for the presence of residual or recurrent carcinoma after hormonal therapy for cancer. PMID- 17134737 TI - The higher incidence of squamous cell carcinoma in renal transplant recipients is associated with increased telomere lengths. AB - The incidence and aggressiveness of nonmelanoma skin cancers, including basal cell carcinoma and squamous cell carcinoma (SCC), in immunocompromised renal transplant recipients (RTRs) is dramatically higher (up to 100-fold) compared with the normal population. SCC lesions are also predominant in RTRs, in contrast to the normal population where basal cell carcinoma is more common. The mechanisms underlying this phenomenon are unknown, but effective treatments for these skin tumors would have a significant impact upon morbidity in this group of patients. The fundamental role of telomeres and telomerase in the development of most human cancers, including melanoma, is well established, but very few reports have assessed their function during the onset of nonmelanoma skin cancer. To assess whether telomere maintenance plays any role in the increased incidence of SCC in renal transplant patients, we analyzed both the telomere lengths and telomerase expression levels in 44 SCCs and 22 Bowen's disease (BD) samples (carcinoma in situ) from RTRs and nontransplant patients. Our findings provide statistically significant evidence that the telomeres are consistently longer in both BD RTR and SCC RTR lesions compared with their nontransplant counterparts. We also show by immunohistochemistry that there is a trend toward higher telomerase levels in both the BD RTR and SCC RTR lesions, although this was not statistically significant. Our data thus suggest that telomere lengthening may possibly be an early event in the development of SCC in renal transplant patients and demonstrate that telomere maintenance mechanisms should be further evaluated with respect to developing a future therapeutic strategy for these cancers. PMID- 17134738 TI - Immunohistochemistry of primary malignant neuroepithelial tumors of the kidney: a potential source of confusion? A study of 30 cases from the National Wilms Tumor Study Pathology Center. AB - Ewing sarcoma/peripheral primitive neuroectodermal tumor (pPNET) is a rare primary tumor of the kidney with morphologic features similar to those of other primitive tumors. Previous studies have shown that these tumors frequently stain positively with immunostains against CD99 and FLI-1 and negatively with stains against WT-1, suggesting that these markers may be used for the distinction between Wilms tumor and pPNET. We present 30 cases of primary malignant neuroepithelial tumor with immunohistochemical profiles and reverse transcriptase polymerase chain reaction (RT-PCR) analysis and show that immunophenotypic overlap exists between Wilms tumor and pPNET. A subset of 30 neuroepithelial tumors from the National Wilms Tumor Study originally categorized as putative pPNETs of the kidney was stained with FLI-1, WT-1, and thyroid transcription factor-1. Bicolor fluorescence in situ hybridization studies were performed on 19 of the cases. Other data on these tumors were available from a previous study (Am J Surg Pathol 2001;25:133). Of 7 primary tumors that had the EWS/FLI-1 fusion transcript by RT-PCR, 6 exhibited strong immunopositivity for FLI-1. Nine that were negative by RT-PCR stained positively with the FLI-1 stain. Five fusion negative cases stained with both FLI-1 and WT-1. Three fusion-negative cases were negative for FLI-1 but positive for WT-1. Five fusion-negative cases were negative for both FLI-1 and WT-1. Of the 30 cases, 29 were positive for CD99. Seven cases that were negative for the EWS-FLI-1 fusion by RT-PCR were positive by fluorescence in situ hybridization. All cases were negative for thyroid transcription factor-1. Reliance upon immunohistochemistry as the sole means of ancillary diagnosis in renal pPNET can lead to confusing results. We recommend molecular fusion studies for clarification of primitive renal tumors with unexpected immunophenotypic results. PMID- 17134739 TI - Composite splenic marginal zone lymphoma and mantle cell lymphoma arising from 2 independent B-cell clones. AB - We report the first case of composite lymphoma involving both mantle cell lymphoma (MCL) and splenic marginal zone lymphoma (SMZL) with circulating villous lymphocytes. Morphological, immunohistochemical, immunophenotyping, as well as detailed genetic studies (fluorescence in situ hybridization, IGVH gene sequencing), were performed and confirmed the existence of 2 independent, unrelated tumor clones. The MCL component expressed IgMD lambda, was CD5+, harbored a t(11;14)(q13;q32) involving CCND1, and showed an unmutated VH1-18 gene rearrangement. The SMZL component expressed IgMD kappa, was CD5-, showed a t(10;14)(q24;q32) and an unmutated VH3-7 gene rearrangement. Interestingly, this t(10;14) targeted the NFKB2 gene. Only a single other case of SMZL with t(10;14)/NFKB2 has been reported. Taken together, these data indicate that the MCL and SMZL arose as a consequence of independent malignant transformation events within an antigen-naive B-cell population. This case highlights the importance of a multidisciplinary approach and tissue diagnosis in these complex situations. PMID- 17134740 TI - Resident physician opinions on autopsy importance and procurement. AB - The national decline in hospital autopsy cases negatively impacts physician education and medical quality control to an unknown degree. The current non medicolegal autopsy rate is less than 5% of hospital deaths. This study compares internal medicine and pathology resident physician perceptions of the autopsy, including the importance, procurement, technique, and the pathologist-internist interaction. An 84-item survey based on autopsy literature was designed, piloted, and distributed to 214 residents at a single 800+ bed tertiary care academic teaching hospital (Massachusetts General Hospital, Boston) to accomplish this goal. Completed surveys were obtained from 72% of medicine (n = 118) and 84% of pathology (n = 42) residents. Residents strongly agree on the importance of autopsies for education, answering clinical questions, public health, and research. Autopsy rates are deemed inadequate. Internists are comfortable requesting autopsies, but report insufficient guidance and difficulty with answering technical questions. Although not requested on all hospital deaths, internists are more likely to initiate an autopsy request than a decedent's family, and worry significantly less about institutional costs and malpractice litigation than pathologists believe. Internists expressed interest in having an instructional brochure to give families, observing an autopsy, and having increased communication and support with autopsies from pathology residents. The main reasons why autopsy consent is not requested (it is unpleasant, cause of death is known, family is upset or seems unwilling) and why families refuse (patient has suffered enough, body may be handled disrespectfully, religious/moral objections, lack information) were similar for both resident groups. Despite their decline, autopsies still remain important to medicine as indicated by internal medicine and pathology residents at a large academic center. Improving autopsy education, enhancing availability of resources, and strengthening the pathologist-internist collaboration may serve to heighten awareness and ultimately procurement. PMID- 17134741 TI - An ageing question: do embryonic stem cells protect their genomes? AB - Many physiological and cellular processes contribute to the ageing of individuals. One hypothesis argues that the genomes of somatic cells accumulate mutations, which, in turn, alter the metabolism of the cells and contribute to the ageing process. The frequency of somatic mutation approaches 10(-4) and the majority of mutagenic events at heterozygous loci is due to loss of heterozygosity as a consequence of mitotic recombination. A corollary to the argument that somatic cells accumulate mutations is that cells of the germ line and ES cells have a greater requirement for maintaining the integrity of their genomes. In the former case, a high somatic mutation frequency predicts an increase in somatic disease, which limits our lifespan. The corollary is that cells of the germline and ES cells must minimize the mutational burden to limit the frequency of congenital disease and to ensure the proper transmission of undamaged DNA to the gene pool. This report describes two mechanisms utilized by murine ES cells to minimize DNA damage within the proliferative pool. In the first case, murine ES cells display a frequency of mutation and mitotic recombination that is about 100-fold lower than that observed in somatic cells. Second, ES cells lack a G1 checkpoint following DNA damage. When subjected to ionizing radiation, the fraction of apoptotic cells increases to about 40%. Ectopic expression of Chk2 is sufficient to establish a G1 arrest and the concomitant protection from cell death. PMID- 17134742 TI - The molecular basis of ageing in stem cells. AB - Ageing is often defined in the context of telomerase activity and telomere length regulation. Most somatic cells have limited replication ability and undergo senescence eventually. Stem cells are unique as they possess more abundant telomerase activity and are able to maintain telomere lengths for a longer period. Embryonic stem cells are particularly resistant to ageing and can be propagated indefinitely. Remarkably, adult somatic cells can be reprogrammed to an ESC-like state by various means including cell fusion, exposure to ESC cell free extracts, enforced expression of specific molecules, and somatic cell nuclear transfer. Thus, the rejuvenation of an 'aged' state can be effected by the activation of specific key molecules in the cell. Here, we argue that cellular ageing is a reversible process, and this is determined by the balance of biological molecules which directly or indirectly control telomere length and telomerase activity, either through altering gene expression and/or modulating the epigenetic state of the chromatin. PMID- 17134743 TI - On the role of placental growth factor in murine adipogenesis. AB - The potential role of placental growth factor (PlGF) in early stages of adipogenesis was investigated in vivo using a murine model of obesity, as well as in vitro using cultured preadipocytes. PlGF-deficient (PlGF-/-) and wild-type (WT) mice, kept on high fat diet (HFD) for 3 weeks, had comparable body weight and weight of subcutaneous (SC) and gonadal (GON) adipose tissues. Blood vessel size and blood vessel density, normalized to adipocyte number, were not significantly different in SC and GON adipose tissues of both genotypes. Differentiation of embryonic fibroblasts derived from WT or PlGF-/- mice into mature adipocytes was comparable. Furthermore, addition of recombinant PlGF, of the PlGF neutralizing MAb PL5D11D4 or of the anti-Flk-1 MAb DC101 to cultured 3T3 F442A preadipocytes did not significantly affect their differentiation into mature adipocytes. Ex vivo blood vessel outgrowth following seeding of adipose tissue-derived microvessel fragments in 3D-collagen gels was not affected by PlGF deficiency. Thus, in murine model systems, PlGF does not seem to play an important role in early adipogenesis. PMID- 17134744 TI - P-glycoprotein ATPase activating effect of opioid analgesics and their P glycoprotein-dependent antinociception in mice. AB - It is well known that opioid analgesics exert central antinociceptive actions. However, in vivo and in vitro studies have shown that some opioid analgesics given systemically have limited access to the central nervous system because of the blood-brain barrier (BBB). P-glycoprotein (P-gp), an ATP-dependent drug efflux transporter, is one component of the BBB. In this report, we assessed the antinociceptive effect of morphine, fentanyl, and meperidine in P-gp deficient (mdr1a KO) mice, and compared these effects with those in wild type (WT) mice. The antinociceptive effects of morphine and fentanyl in mdr1a KO mice were significantly greater than those in WT mice. However, there was no clear difference in the antinociceptive effects of meperidine in the two genotypes. In addition, we determined the effect of opioid analgesics on P-gp ATPase activity, which is requisite for drug transport, using mouse brain capillary endothelial cells. In our observations, morphine and fentanyl, but not meperidine, significantly increased P-gp ATPase activity, and the drugs' concentration response curves were bell-shaped, reaching a peak at a concentration of 1 muM. These results suggest that P-gp ATPase activity may be, at least in part, involved in the antinociceptive potencies of those opioid analgesics that are substrates for P-gp. PMID- 17134745 TI - Armodafinil, the R-enantiomer of modafinil: wake-promoting effects and pharmacokinetic profile in the rat. AB - Modafinil reduces the excessive sleepiness associated with narcolepsy, obstructive sleep apnea/hypopnea syndrome, and shift work sleep disorder. In rats, modafinil promotes dose-dependent increases in wake duration. The wake promoting activity of the R-enantiomer of modafinil (armodafinil) was evaluated in WKY rats and compared to the classical stimulant, D-methamphetamine. Electroencephalographic and electromyographic signals were assessed via a tethered cranial implant. Body temperature and locomotor activity were assessed by telemetry via intraperitoneal implant. Rats (n=60, 12 per group) were subjected to one of five parallel treatments: armodafinil at 30, 100 and 300 mg/kg i.p.; D-methamphetamine, 1 mg/kg i.p. and vehicle. Armodafinil and D methamphetamine increased time spent awake relative to vehicle. Armodafinil evoked increases in wake duration were dose-dependent and proportional to plasma compound exposure. Induction of wakefulness by D-methamphetamine was associated with an approximately two-fold increase in locomotor activity during the 2-h period immediately following administration relative to vehicle. D methamphetamine also increased body temperature over the same time interval. The dose of armodafinil (100 mg/kg, i.p.) that was closest to D-methamphetamine in its wake-promoting efficacy did not produce changes in either body temperature or the intensity of locomotor activity relative to vehicle. Acute rebound hypersomnolence, characterized by increases in non-rapid eye movement sleep (NREMS) as a percentage of time and NREMS bout duration and by a decreased frequency of brief awakenings following sleep deprivation, occurred following D methamphetamine-but not armodafinil-induced wake in this rat model which has been shown to be predictive of human drug responses. PMID- 17134746 TI - Parthenogenetic activation of domestic cat oocytes using ethanol, calcium ionophore, cycloheximide and a magnetic field. AB - The objective of this study was to evaluate parthenogenetic activation of domestic cat oocytes after being exposed to either ethanol, magnetic field, calcium ionophore A23187, or cycloheximide and a combination of these agents. We also wished to evaluate the usefulness of the magnetic field for oocyte activation. In vitro matured oocytes subjected to artificial activation were randomly assigned into eight groups according to activating agents: (1) 10% ethanol; (2) the magnetic field (slow-changing, homogenous magnetic field with low values of induction); (3) 10% ethanol plus magnetic field; (4) 10 microM calcium ionophore A23187; (5) 10 microM calcium ionophore A23187 plus magnetic field; (6) 10% ethanol and 10 microg/mL of cycloheximide; (7) 10% ethanol and 10 microg/mL of cycloheximide plus magnetic field; (8) oocytes were not exposed to any of the activating agents. After activation oocytes were stained with Hoechst 33258 and parthenogenetic activation was defined as oocytes containing pronuclei and second polar bodies or two to four or six nuclei (embryonic cleavage). The total activation rate by using different activation treatments was 40%. The addition of the magnetic field to ethanol or calcium ionophore treatments resulted in increased parthenogenetic activation rates from 47% to 75%, and from 19% to 48%, respectively (P<0.001). Instead, when the magnetic field was added to ethanol and cycloheximide treatment, activation rate decreased from 48% to 30%. Oocytes activated with magnetic field only gave the lowest activation rate (12%). We concluded that a magnetic field can be used as an activating agent, and the combination of ethanol and magnetic field is an effective method for domestic cat oocyte activation. PMID- 17134748 TI - Preparation of mica-based glass-ceramics with needle-like fluorapatite. AB - OBJECTIVES: The aim of this study was to prepare mica-based glass-ceramics containing needle-like fluorapatites, and to understand the relationship between the composition, the microstructure and mechanical properties. METHODS: The specimens were prepared by casting and subsequent heat treatment. The crystalline phases in the specimens were determined by X-ray diffraction (XRD). The microstructures of the fractured surface were examined by using a scanning electron microscope with an energy dispersive spectrometer (SEM/EDS). The Vickers hardness and fracture toughness were determined by the indentation method. Statistical analysis of the results was performed by one-way analysis of variance (ANOVA) and Tukey's test. The bioactivity of the glass-ceramics was evaluated by soaking the polished specimens in the simulated body fluid (SBF). RESULTS: Needle like fluorapatite crystals were successfully obtained in mica-based glass ceramics by controlling the heat treatment process. The formation of needle-like fluorapatite crystals, instead of particle-like crystals reported in previous studies, can be attributed to the one-dimensional rapid growth of fluorapatite along the c-axis. The mechanical properties of the glass-ceramics are related to the fluorapatite content. The higher the fluorapatite content, the higher the Vickers hardness and the fracture toughness (p<0.01). The bioactivity test showed that the needle-like fluorapatite-containing glass-ceramics possess very good bioactivity. SIGNIFICANCE: As the needle-like fluorapatite crystal has the same morphology as human bones, it endows the mica-based glass-ceramics with good bioactivity. Moreover, the needle-like fluorapatite crystal toughens the base glass-ceramics. Therefore, the mica-based glass-ceramics with needle-like fluorapatite have the potential for the restoration of bone defects and implants. PMID- 17134749 TI - Engineering of bone using bone marrow stromal cells and a silicon-stabilized tricalcium phosphate bioceramic: evidence for a coupling between bone formation and scaffold resorption. AB - Resorbable porous ceramic constructs, based on silicon-stabilized tricalcium phosphate, were implanted in critical-size defects of sheep tibias, either alone or after seeding with bone marrow stromal cells (BMSC). Only BMSC-loaded ceramics displayed a progressive scaffold resorption, coincident with new bone deposition. To investigate the coupled mechanisms of bone formation and scaffold resorption, X-ray computed microtomography (muCT) with synchrotron radiation was performed on BMSC-seeded ceramic cubes. These were analyzed before and after implantation in immunodeficient mice for 2 or 6 months. With increasing implantation time, scaffold thickness significantly decreased while bone thickness increased. The muCT data evidenced that all scaffolds showed a uniform density distribution before implantation. Areas of different segregated densities were instead observed, in the same scaffolds, once seeded with cells and implanted in vivo. A detailed muX-ray diffraction analysis revealed that only in the contact areas between deposited bone and scaffold, the TCP component of the biomaterial decreased much faster than the HA component. This event did not occur at areas away from the bone surface, highlighting coupling and cell-dependency of the resorption and matrix deposition mechanisms. Moreover, in scaffolds implanted without cells, both the ceramic density and the TCP:HA ratio remained unchanged with respect to the pre-implantation analysis. PMID- 17134750 TI - Multidrug resistance protein expression of adult T-cell leukemia/lymphoma. AB - In adult T-cell leukemia/lymphoma (ATL), it is difficult to achieve remission and the reason for the resistance to chemotherapeutic agents may be linked to the presence of multidrug resistance (MDR) proteins. Lung resistance-related protein (LRP), multidrug resistance-associated protein and P-glycoprotein are three MDR proteins which we examined in ATL cells using multiparametric flow cytometry and real-time RT-PCR. LRP was highly expressed and suppressing LRP function increased doxorubicin accumulation in nuclei. This indicates LRP may be contributing to drug resistance in ATL patients, and the suppression of LRP function could be a new strategy for ATL treatment. PMID- 17134751 TI - Severe pulmonary complications after bortezomib treatment for multiple myeloma: an unrecognized pulmonary vasculitis? PMID- 17134752 TI - Cultural factors in decision-making about child physical abuse: identifying reporter characteristics influencing reporting tendencies. AB - OBJECTIVE: This study examined cultural factors that may influence child physical abuse reporting. Relevant cultural factors such as the respondents' ethnic identity and corporal punishment acceptability were investigated as proximal variables of ethnicity that affect child physical abuse reporting tendencies. METHOD: Participants consisted of 378 Caucasian, Hispanic, and African American college students. Participants read 12 vignettes that varied the ethnicity of the child and described a parent using physical punishment with the child. Participants' level of ethnic identity and their acceptance of different discipline practices were also assessed. Data were analyzed using multiple regression procedures. RESULTS: The ethnicity of the respondent was a significant predictor of reporting tendencies for African American respondents only. For African Americans, higher levels of ethnic identity and acceptance of corporal punishment were significant mediators of reporting tendencies, which influenced the relationship between ethnicity and likelihood to report a child of the same ethnicity. CONCLUSION: The results of this study indicate that ethnic minorities may not share similar cultural values and may differ in their degree of adherence to certain cultural variables. These results demonstrate how cultural variables that influence reporting tendencies may not apply across ethnic groups. Therefore, specifying relevant cultural variables is essential to understanding the relationship between ethnicity and reporting tendencies. PMID- 17134753 TI - Adenosine monophosphate-induced amplification of ColE1 plasmid DNA in Escherichia coli. AB - ColE1 plasmid copy number was analyzed in relaxed (relA) and stringent (relA(+)) Escherichia coli cells after supplementation of culture media with adenosine monophosphate (AMP). When a relaxed E. coli strain bearing ColE1 plasmid was cultured in LB medium for 18 h and induced with AMP for 4h, the plasmid DNA yield was significantly increased, from 2.6 to 16.4 mgl(-1). However no AMP-induced amplification of ColE1 plasmid DNA was observed in the stringent host. Some plasmid amplification was observed in relA mutant cultures in the presence of adenosine, while adenine, ADP, ATP, ribose, potassium pyrophosphate and sodium phosphate caused a minor, if any, increase in ColE1 copy number. A mechanism for amplification of ColE1 plasmid DNA with AMP in relA mutant bacteria is suggested, in which AMP interferes with the aminoacylation of tRNAs, increases the abundance of uncharged tRNAs, and uncharged tRNAs promote plasmid DNA replication. According to this proposal, in relA(+) cells, the AMP induction could not increase ColE1 plasmid copy number because of lower abundance of uncharged tRNAs. Our results suggest that the induction with AMP can be used as an effective method of amplification of ColE1 plasmid DNA in relaxed strains of E. coli. PMID- 17134754 TI - Violent and sexual offences: a validation of the predictive quality of the PCL:SV in Switzerland. AB - Used to diagnose psychopathy, the Psychopathy Checklist: Screening Version (PCL:SV) is still rarely used in the German-speaking part of Europe. The aim of the present study was to conduct a first-time evaluation of the predictive validity of the PCL:SV for sexual and violent recidivism in the German language area. PCL:SV scores for 96 violent and sexual offenders in Switzerland were assessed. These assessments were based on data taken from the forensic psychiatric expert opinions. The scores were then compared to subsequent recidivism as shown in the official criminal records. ROC analysis indicated a cut-off score of 14 for maximized overall predictive accuracy. Consistent with various results from past research in North America and Europe, the PCL:SV yielded a satisfying predictive accuracy (ROC area=.69). This degree of accuracy demonstrates the PCL:SV's usefulness for risk assessment of sexual and violent delinquency in the Swiss cultural and German language area. PMID- 17134755 TI - FcRgamma chain does not replace CD3zeta chain in CD3zeta-deficient T lymphocytes of patients with gastric adenocarcinoma. AB - Defective CD3zeta chain expression has been reported in T lymphocytes of patients with inflammatory diseases, such as systemic lupus erythematosus or osteoarthritis, and with cancer. In lupus, the absent CD3zeta chain is replaced by the FcRgamma chain, rendering the T cells hyper responsive. However, there are no data on T lymphocytes from patients with cancer. In this study, the presence of the FcRgamma chain and its associated kinase, Syk, was analysed in patients with gastric adenocarcinoma and healthy subjects. Western blot and immunoprecipitation experiments were carried out with total cell or lipid raft extracts from fresh peripheral blood mononuclear cells or T lymphocytes, and Herpesvirus saimiri-derived T-cell lines (of blood or tissue origin). Our results revealed that the absent CD3zeta chain in cancer T lymphocytes was not replaced by FcRgamma either in fresh T cells or T-cell lines, in contrast to lupus T cells. This altered expression of signalling molecules in T lymphocytes of cancer patients, would explain their low proliferative capacity. Our T-cell lines represent tools to unveil the signalling abnormalities of cancer T lymphocytes. PMID- 17134756 TI - A novel pentamer versus pentamer approach to generating neutralizers of verotoxin 1. AB - Verotoxins (VTs), or shiga-like toxins, are produced by enterohemorrhagic Escherichia coli (EHEC), which cause hemorrhagic colitis and hemolytic uremic syndrome. VTs are the major virulence factors in EHEC infection due to their cytotoxicity to various types of cells. Here, we present a novel type of VT neutralizer based on pentavalent single-domain antibodies, or pentabodies. Two single-domain antibodies (sdAbs) specific for the receptor binding sites of the B subunit of VT1 (VT1B) were isolated from a naive llama phage display library. These two sdAbs were pentamerized to generate pentameric VT neutralizers, VTI-1 and VTI-3. Both VT neutralizers bound wild type VT1B specifically with superior functional affinity. In vitro neutralization assays showed that VTI-1 and VTI-3 were able to neutralize 90% and 40%, respectively, of the cytotoxicity caused by VT1. This effort provides the basis of a novel type of VT neutralizer that can potentially be produced at a relatively low cost. PMID- 17134757 TI - Direct impact of inactivated HIV-1 virions on B lymphocyte subsets. AB - Although there is no convincing evidence that HIV infects primary B cells, marked changes in B cell responses have been described in HIV-1-infected subjects, including B cell repertoire perturbations, depression of B cell memory and paucity of CD5(+) B cells. As it is hard to assess the consequences of these in vitro and ex vivo observations in patients, the pathogenic mechanisms responsible for the B cell deficit are unclear, and direct and indirect effects of HIV-1 remain possible. To gain further insight into the impact of HIV-1 on the B cell compartment in vivo, we used XenoMouse mice, mice genetically engineered to express human antibodies with an absence of mouse antibody expression. In these transgenic animals, B cells expressing a virtually full human Ig repertoire develop, which allows investigation of the in vivo consequences of confronting B cells expressing human immunoglobulins with HIV-1. We found that soluble gp120 induced an inversion in the B-1a/B-1b cell ratios, without impacting B-2 cells or affecting substantially the T cell compartment. Virion treatment specifically and dramatically depressed B-1a cells, which represent the majority of B-1 cells in normal mice. The observed B cell changes were associated with a functional alteration of the humoral response to tetanus toxoid. Thus, the results reveal a capacity of HIV-1 to specifically impact a highly specialized B cell subpopulation. Because there is evidence that human IgM memory B cells are functionally equivalent to murine B-1a cells, our findings suggest that gp120 may have a direct deleting activity on B cell memory. PMID- 17134758 TI - Breakdown kinetics of the tri-chromium(III) oxo acetate cluster ([Cr(3)O(OAc)(6)]+) with some ligands of biological interest. AB - Kinetics for the breakdown of the trinuclear chromium acetate cluster, [Cr(3)O(OAc)(6)](+), with a series of monoprotic and diprotic ligands in weakly acidic aqueous media (pH approximately 4 or approximately 5) have been investigated spectrophotometrically at 40-60 degrees C. The results point to an ion-pair equilibrium as the first step followed by associative interchange mechanism forming the mononuclear product of the reaction. Pseudo-first-order rates were determined from absorbance data and associated activation parameters were calculated using the Eyring equation. Enthalpy and entropy terms of the reactions (e.g., histidine, DeltaH(double dagger) = 75 +/- 15 kJ mol(-1), DeltaS(double dagger) = -130 +/- 25 J K(-1) mol(-1); lactic acid, DeltaH(double dagger) = 66 +/- 13 kJ mol(-1), DeltaS(double dagger) = -155 +/- 30 J K(-1) mol( 1); glycine, DeltaH(double dagger) = 31 +/- 6 kJ mol(-1), DeltaS(double dagger) = -225 +/- 45 J K(-1) mol(-1)) are consistent with an associative interchange (I(a)) mechanism, and produce a linear isokinetic plot (slope = 50 degrees C). Rates and activation parameters are comparable to those of substitution reactions of the chromium(III) hexaaqua cation. Other ligands studied included malonic acid and the amino acid, aspartic acid. Observed rates are faster than water exchange rates, but typically slower than anion substitution rates, and indicate that trinuclear chromium(III) clusters are expected to be kinetically stable in neutral to slightly acidic conditions. PMID- 17134759 TI - Relatively stable N-ligated [2Fe2S](2+) clusters with dipyrromethane capping ligands. AB - [2Fe2S] clusters with terminal N-ligation (His, Arg) and unique functions are increasingly recognized in biological systems. In this work three new [2Fe2S] clusters 1-3 with different 1,1'-dipyrrolmethane derivatives as chelating terminal ligands have been prepared and fully characterized, including by X-ray crystallography: (NEt(4))(2)[L(2)Fe(2)(mu-S)(2)] with L=Me(2)C(C(4)H(3)N)(2) (1), Ph(2)C(C(4)H(3)N)(2) (2), (CH(2))(5)C(C(4)H(3)N)(2) (3). These systems represent rare examples of synthetic [2Fe2S] complexes with N-donor capping ligands. While geometric parameters as well as spectroscopic and electrochemical characteristics of the new complexes are as anticipated, the chelating nature of the terminal ligands in 1-3 imparts a relatively high stability that will be advantageous for reactivity studies of the [2Fe2S] core. PMID- 17134760 TI - A dense array stimulator to generate arbitrary spatio-temporal tactile stimuli. AB - The generation and presentation of tactile stimuli presents a unique challenge. Unlike vision and audition, in which standard equipment such as monitors and audio systems can be used for most experiments, tactile stimuli and/or stimulators often have to be tailor-made for a given study. Here, we present a novel tactile stimulator designed to present arbitrary spatio-temporal stimuli to the skin. The stimulator consists of 400 pins, arrayed over a 1cm(2) area, each under independent computer control. The dense array allows for an unprecedented number of stimuli to be presented within an experimental session (e.g., up to 1200 stimuli per minute) and for stimuli to be generated adaptively. The stimulator can be used in a variety of modes and can deliver indented and scanned patterns as well as stimuli defined by mathematical spatio-temporal functions (e.g., drifting sinusoids). We describe the hardware and software of the system, and discuss previous and prospective applications. PMID- 17134761 TI - Chronically recording with a multi-electrode array device in the auditory cortex of an awake ferret. AB - It is known that anesthesia depresses neural activity and inhibits cortico cortical interactions and cortical output. Hence, it is important to record from awake animals in order to better understand the full dynamic range of neural responses. We have developed a preparation for chronic, multi-electrode physiological recording in the cortex of the awake ferret. This paper discusses several of the advantages and disadvantages of the technique as well as procedures used to overcome potential complications associated with chronic implants in the ferret. Our solutions are well suited to the special species requirements, yet are also easily generalizable to other species. PMID- 17134762 TI - Tryptophan hydroxylase 1 gene (TPH1) moderates the influence of social support on depressive symptoms in adults. AB - BACKGROUND: Tryptophan hydroxylases (TPHs) are involved in the biosynthesis of serotonin and are therefore candidate genes for psychiatric disorders, including depression. We examined whether the common 218 A > C and 779 A > C polymorphisms in the tryptophan hydroxylase 1 gene (TPH1) moderated the association between perceived social support and sub-clinical depressive symptoms in adults. METHODS: The subjects were a randomly selected subsample (n=341) of individuals participating in the Cardiovascular Risk in Young Finns study, who had data on social support on one assessment time and depressive symptoms on two assessment times. Social support was assessed on the Perceived Social Support Scale Revised (PSSS-R) and depressive symptoms on a modified version of the Beck's Depression Inventory (BDI). RESULTS: We found that low social support predicted depressive symptoms more strongly in individuals carrying A alleles of the TPH1 than in others. The interaction effect was observed in a cross-sectional analysis and when predicting depressive symptoms over a four-year period. LIMITATIONS: We did not have data on TPH2, which has recently been identified as the primary TPH isomorphism affecting serotonin synthesis in the brain. CONCLUSIONS: TPH1 gene may be involved in the development of depressive symptoms by moderating the impact of depressogenic social influences. PMID- 17134763 TI - Response styles to depressed mood in bipolar affective disorder. AB - BACKGROUND: It has been hypothesised that dysfunctional strategies for avoiding depression play an important role in the pathway to mania. Support for this hypothesis comes from studies demonstrating that remitted and manic bipolar patients show similar cognitive biases to currently depressed patients. METHOD: Manic patients, depressed bipolar patients, remitted bipolar patients and healthy controls were compared on an expanded version of Nolen-Hoeksema's [Nolen Hoeksema, S., 1991. Responses to depression and their effects on the duration of depressed mood. Journal of Abnormal Psychology, 100, 569-582.] Response Styles Questionnaire, measuring strategies for coping with depression. RESULTS: Manic patients reported greater use of active-coping and risk-taking compared to the depressed, remitted and healthy controls. Bipolar remitted patients reported greater rumination. CONCLUSIONS: The findings are consistent with the hypothesis that mania is associated with dysfunctional strategies for regulating negative emotion. PMID- 17134764 TI - Controlling autoimmunity--Lessons from the study of red blood cells as model antigens. AB - The characterization of human and animal red blood cell (RBC) autoantigens in autoimmune hemolytic anemia (AIHA) has provided an opportunity study the control of specific autoimmune responses of unequivocal pathogenic relevance. The results reveal that censorship of the autoimmune helper T (Th) cell repertoire by deletion and anergy is very incomplete in healthy individuals, even for widely distributed, abundant self-antigens on RBC. There is strong evidence that autoaggression by surviving Th cells is normally held in check by other mechanisms, including failure to display the epitopes that they recognize, and active immunoregulation. AIHA is one of the first human autoimmune diseases in which regulatory T (Tr) cells that are specific for the major autoantigens have been identified. These Tr cells recognize the dominant naturally processed epitopes, and recent studies suggest that disease develops when other determinants, to which such tolerance is less secure, and which are normally inefficiently presented, are displayed at higher levels. Together, the results raise the possibility that therapy for diseases such as AIHA could be based on switching the balance of the response back towards regulation, in particular by the administration of the dominant peptides recognized by specific Tr cells. PMID- 17134765 TI - Histological analysis of palatopharyngeal muscle from children with snoring and obstructive sleep apnea syndrome. AB - Obstructive sleep apnea syndrome (OSAS) is an upper airway obstruction that occurs during the sleep. One of the suggested mechanisms involved in this process is a neuromuscular abnormality of the palatal muscles. Whether children with OSAS develop into OSAS adults, or children and adult OSAS are two distinct disorders occurring at different ages are questions to be answered. Here, we made the histological analysis of palatophryngeal muscle in 34 oral-breathing children of both genders, aged 5-12 years old, with hypertrophic tonsils and adenoids. According to the polysomnographic study the participants were divided into children without sleeping disorders (group I) and children with primary snoring (group II) or apnea (group III). The main histological findings were fiber size variability in 70% cases from groups II and III and in 71% from group I; perimysial connective tissue infiltration in 48% children from groups II and III and in 71% from group I; intracytoplasmatic mitochondrial proliferation in 63% cases from groups II and III and in 57% cases from group I. Muscle necrosis was only observed in one case, in association with subglandular inflammation. Others findings observed in all groups included fibers with internal architecture alteration, such as moth-eaten and lobulated fibers, type 2 fiber predominance, and small areas of fiber type grouping. The presence of similar histological findings in the palatopharyngeal muscle in children with primary snoring or apnea but also in children without sleeping disorders indicate that such changes could be a normal histological feature of this muscle rather than a neurogenic or myopathic pathology. PMID- 17134766 TI - Different real-time PCR assays could lead to a different result of detection of varicella-zoster virus in facial palsy. AB - Real-time PCR is a useful tool for rapid detection of viral genomic DNA. However, there are many types of real-time PCR, and this variation may induce different results. The sensitivity of two different real-time PCR assays was evaluated for the detection of the varicella-zoster virus (VZV) genome: LightCycler PCR and TaqMan PCR. Auricular skin cells and saliva were sampled from 201 patients with facial nerve paralysis. A hundred and seventy-one of these patients were diagnosed clinically with Bell's palsy, and the remaining 30 with Ramsay Hunt syndrome. In 30 specimens obtained from Ramsay Hunt syndrome patients, VZV DNA was detected in 26 skin and 3 saliva specimens using the LightCycler PCR, while 28 skin and 9 saliva specimens were positive using the TaqMan PCR. None of the patients with Bell's palsy were positive for VZV by the LightCycler PCR, whereas five of these patients were positive by the TaqMan PCR. The TaqMan PCR assay has a better sensitivity compared to the LightCycler PCR for the detection of VZV genome from patients with facial palsy. Further study is required to develop a more sensitive real-time PCR. PMID- 17134767 TI - Nur77 and retinoid X receptors: crucial factors in dopamine-related neuroadaptation. AB - Dopaminergic systems in the brain adapt in response to various stimuli from the internal and external world, but the mechanisms underlying this process are incompletely understood. Here, we review recent evidence that certain types of transcription factor of the nuclear receptor family, specifically Nur77 and retinoid X receptors, have important roles in adaptation and homeostatic regulation of dopaminergic systems. These findings call for a reassessment of our fundamental understanding of the molecular and cellular basis of dopamine mediated transmission. Given that diseases such as Parkinson's disease and schizophrenia are thought to involve adaptation of dopamine signalling, these findings might provide new insight into these pathologies and offer new avenues for drug development. PMID- 17134768 TI - Genetic exchange in Trypanosoma brucei: evidence for mating prior to metacyclic stage development. PMID- 17134769 TI - Sympathetic and hyperthermic reactions by orexin A: role of cerebral catecholaminergic neurons. AB - This experiment tested the effect of a lesion of cerebral catecholaminergic neurons on the sympathetic and thermogenic effects induced by an intracerebroventicular (icv) injection of orexin A. The firing rates of the sympathetic nerves to the interscapular brown adipose tissue (IBAT), along with IBAT, colonic temperatures and heart rate were monitored in urethane-anesthetized male Sprague-Dawley rats before an injection of orexin A (1.5 nmol) into the lateral cerebral ventricle and over a period of 150 min after the injection. Three days before the experiment, the rats were pre-treated with an icv injection of 6-hydroxydopamine (6-OHDA) or 6-OHDA plus desipramine or saline. The results show that orexin A increases the sympathetic firing rate, IBAT, colonic temperatures and heart rate in the rats pre-treated with saline. This increase is blocked by the pre-treatment with 6-OHDA alone or 6-OHDA plus desipramine. These findings indicate that cerebral catecholaminergic neurons (particularly the dopaminergic pathway) play a fundamental role in the complex reactions related to activation of the orexinergic system. PMID- 17134770 TI - Effect of free fatty acid inhibition on silent and symptomatic myocardial ischemia in diabetic patients with coronary artery disease. AB - OBJECTIVE: Free fatty acid inhibition with trimetazidine (TMZ) improves myocardial metabolism and myocardial ischemia in patients with coronary artery disease (CAD). Because of its effect on myocardial glucose utilization TMZ may represent a therapeutic option in diabetic patients with CAD. Aim of the present study was to evaluate whether the metabolic effect of TMZ may improve episodes of myocardial ischemia in diabetic patients with CAD. RESEARCH DESIGN AND METHODS: We assessed the effect of TMZ on 24 h ambulatory ECG monitoring (AEM) in 30 patients (22 males and 8 females, mean (SE) age 67+/-6.5 years) with NIDDM and ischemic cardiomyopathy. Patients were randomized to receive on top of standard therapy either TMZ (20 mg, tds) or placebo (tds) and were evaluated at baseline and after 6 months. RESULTS: Patients randomized to TMZ or placebo were comparable regarding demographic data, distribution of CAD, and glicated haemoglobin levels. TMZ significantly reduced the number of episodes of transient myocardial ischemia (-24% compared to baseline, p<0.01; -27% compared to placebo, p<0.01), and Total Ischemic Burden (-28% compared to baseline, p<0.01; -29% compared to placebo, p<0.01). TMZ also significantly reduced the number of silent episodes of myocardial ischemia (-42% compared to baseline and -39% compared to placebo, p<0.01) and the time of silent myocardial ischemia/24 h (-37% compared to baseline and -35% compared to placebo, p<0.01). No significant changes in heart rate were detected between baseline, placebo and TMZ evaluations. CONCLUSIONS: TMZ is effective in reducing silent and symptomatic episodes of transient myocardial ischemia in diabetic patients with CAD on standard anti anginal therapy. PMID- 17134771 TI - Incessant ventricular tachycardia due to subacute thyroiditis. AB - Subacute thyroiditis is a possibly viral, inflammatory thyroid disorder which can cause thyrotoxicosis. Ventricular arrhythmias are uncommon in thyrotoxicosis and usually occur only in those with marked heart failure or associated cardiac disease. In this case, we present a 52-year-old woman having incessant ventricular tachycardia due to subacute thyroiditis without underlying cardiac disease. PMID- 17134773 TI - Isolated noncompaction of the ventricular myocardium in a patient presenting with subarachnoid hemorrhage. PMID- 17134774 TI - Colchicine for the prevention of the postpericardiotomy syndrome: the COPPS trial. AB - Preliminary data have shown that colchicine may be considered for the prevention of the postpericardiotomy syndrome (PPS). The COlchicine for the Prevention of Post-pericardiotomy Syndrome (COPPS) study is a multicenter, double blind, placebo-controlled, randomized trial that will enroll 360 patients after cardiac surgery. The primary end point will be the PSS rate at 12 months. The study will be the first large randomized placebo-controlled trial in this area, and will provide important evidence regarding the possible benefit of colchicine for the primary prevention of the PPS. PMID- 17134775 TI - Double outlet right ventricle demonstrated by multislice computed tomography. AB - A 60 year-old male presented with abnormal systolic heart murmur. Transthoracic echocardiogram revealed right ventricular (RV) hypertrophy and mosaic jet in the RV outflow tract. RV catheterization revealed significantly pressure gradient between the RV and the pulmonary artery. Enhance ECG-gated multislice CT was performed and an axial source image in end-diastole revealed RV myocardial hypertrophy and remarkable hypertrophy of the moderator band. Multiplanar reconstruction images in end-systole revealed hypertrophic anomalous muscle bundles in the RV and significant narrowing of the RV outflow tracts and a post stenotic dilated pulmonary trunk. In volume rendered images, in end-systole, the lumen of RV outflow tract revealed severe stenosis compared with that in end diastole and he was diagnosed with double chamber RV. PMID- 17134776 TI - The Straight Wire Sign: recognizing guidewire exit during percutaneous coronary intervention for chronic total occlusion. AB - Coronary perforation during PCI may be life threatening. Use of stiff wires during instrumentation of totally occluded arteries is a contributing factor. The "Straight Wire" sign presented here was realized from a case of coronary perforation encountered at our cath lab. The identification of this sign in another patient prompted immediate response and avoided further complications. PMID- 17134777 TI - Cardiac hemochromatosis. PMID- 17134778 TI - Speech comprehension assessed by electroencephalography: a new method using m sequence modulation. AB - Electroencephalograms (EEGs) were recorded from eight Japanese speakers while they listened to Japanese and Spanish sentences (approximately 51s each). The sentences were modulated in amplitude by a binary m-sequence and played forward or backward. A circular cross-correlation function was computed between the EEG signals and the m-sequence and averaged across subjects. Independent component analysis of the averaged function revealed a component source response which was obtained only for the comprehensible Japanese and not for the incomprehensible sentences. The present study has thus shown that a 1-min long EEG signal is sufficient for the assessment of speech comprehension. PMID- 17134779 TI - Developmental localization of potassium chloride co-transporter 2 (KCC2) in the Purkinje cells of embryonic mouse cerebellum. AB - Developmental shift in GABA actions from depolarization to hyperpolarization occurs as a result of decreasing the intracellular Cl(-) concentration regulated by K(+)-Cl(-) co-transporter 2 (KCC2). To clarify the time-course of the developmental shift on the Purkinje cells, we examined KCC2-localization in the embryonic mouse cerebellum. The KCC2 was first detected within the Purkinje cells in the Purkinje cell layer of the hemisphere at embryonic day 15 (E15) and the vermis at E17, but the ventricular and intermediate zones were negative. These results suggest that GABA might become inhibitory on the Purkinje cells after their settling in the Purkinje cell layer. PMID- 17134780 TI - Neurokinin-1 and neurokinin-3 receptors in primate substantia nigra. AB - Striatonigral axons co-release GABA and substance P (SP) at their target sites, but little is known about the action of SP at nigral level. Therefore, we studied immunohistochemically the cellular and subcellular localization of SP and its high affinity receptors neurokinin-1 (NK-1R) and neurokinin-3 (NK-3R) at nigral level in squirrel monkeys. Immunofluorescent studies revealed that, although SP+ fibers arborised more densely in the pars reticulata (SNr) than in the pars compacta (SNc), the two nigral divisions harbored numerous neurons expressing NK 1R and NK-3R. Confocal microscopic analyses showed that numerous SNr neurons and virtually all SNc dopaminergic neurons contained both NK-1R and NK-3R. At the electron microscope level, NK-1R and NK-3R were mainly associated with intracellular sites or located at extrasynaptic position on plasma membrane. A small proportion of SP+ boutons also showed NK-3R immunoreactivity. The distribution of NK-1R and NK-3R in SNr and SNc suggests that SP exerts its effect through postsynaptic receptors, as well as via presynaptic autoreceptors and heteroreceptors. These findings indicate that the excitatory peptide SP can modulate the inhibitory action of GABA at nigral level and suggest that the co release of these two neuroactive substances should be taken into account when considering the functional organization of the basal ganglia. PMID- 17134781 TI - Heterogeneity of nitric oxide synthase-containing neurons in the mouse main olfactory bulb. AB - The distribution and structural features of nitric oxide [corrected] synthase (NOS) containing intrinsic neurons were studied in the mouse main olfactory bulb (MOB). NOS positive neurons were heterogeneous, including some subpopulations of periglomerular cells, granule cells, interneurons in the external plexiform layer, superficial and deep short-axon cells and stellate cells. NOS positive periglomerular cells were frequently calretinin immunoreactive and, although rarely, calbindin positive. Importantly, some middle and external tufted cells were also confirmed to be NOS positive, some of which were also cholecystokinin (CCK) positive. Retrograde tracer experiments showed that some NOS positive tufted cells, which were also CCK positive, constitute the intrabulbar association system and the projection system to the olfactory tubercle. In addition, another particular subpopulation of NOS positive neurons with no or little CCK immunoreactivity appeared to project to areas covering the dorsal endopiriform nucleus, claustrum and insular cortex. Furthermore, diverse types of neurons other than mitral/tufted cells were also suggested to be projection neurons of the MOB. The present study revealed the diversity of NOS positive neurons in the mouse MOB and further revealed that they were different from those reported previously in the rat MOB in structural and chemical properties. PMID- 17134782 TI - Immune enhancing effects of Lactobacillus acidophilus LAFTI L10 and Lactobacillus paracasei LAFTI L26 in mice. AB - The immune enhancing properties of Lactobacillus acidophilus LAFTI L10 and Lactobacillus paracasei LAFTI L26 in mice were investigated. Each mouse (BALB/c) was orally fed with cultures of either L. acidophilus or L. paracasei at 10(8) CFU/50 mul per day for 14 days. The effect of these strains on immunoglobulin A, interleukin-10 and interferon gamma producing cells in the gut immune system was determined by immunofluorescence assays. Systemic immune responses were analysed in mice serum upon euthanising after a 14-day feeding trial to estimate cytokines such as IL-10 and IFN-gamma by enzyme-linked immunosorbent assays. L. acidophilus and L. paracasei strains demonstrated an increase in the number of IgA producing cells, IL-10 and IFN-gamma cytokine producing cells in the small intestine. In the systemic immune response, mice fed with L. acidophilus or L. paracasei also enhanced the secretion of anti-inflammatory cytokine (IL-10) and pro-inflammatory cytokine (IFN-gamma). The results of this study suggest that L. acidophilus and L. paracasei were able to enhance specific gut and systemic immune responses in mice. PMID- 17134783 TI - Screening food raw materials for the presence of the world's most frequent clinical cases of Shiga toxin-encoding Escherichia coli O26, O103, O111, O145 and O157. AB - This work aims to provide a strategy for rapidly screening food raw materials of bovine origin for the presence of the most frequent O-serogroups of Shiga toxin encoding Escherichia coli (STEC) involved in food poisoning outbreaks. The prevalence of highly pathogenic serogroups of STEC was surveyed in 25 g portions of minced meat and raw milk using PCR-ELISA and multiplex real-time PCR assays. The prevalence of STEC in raw milk (n=205) and meat samples (n=300) was 21% and 15%, respectively. Contamination by the main pathogenic E. coli O-serogroups representing a major public health concern, including O26, O103, O111, O145, and O157, was potentially around 2.6% in minced meat and 4.8% in raw milk. The MPN values showed an overall contamination ranging from 1 to 2 MPN cells from highly pathogenic serogroups/kg. This survey would indicate that the human pathogenic potential of STEC present in these samples probably remains limited. No conclusion can be drawn at the moment concerning a potential risk for consumers. This rapid screening approach for evaluating the potential presence of highly pathogenic serogroups of STEC in food raw materials should help to improve risk assessment of food poisoning outbreaks. PMID- 17134784 TI - Defective interfering RNAs of Japanese encephalitis virus found in mosquito cells and correlation with persistent infection. AB - Defective interfering (DI) RNAs are deletion mutants of viral genomes that are known in many cases to contribute to persistent infection and modification of viral pathogenesis. Cell type also plays a critical role in the establishment of viral persistence. In this study we have identified for the first time the generation of DI RNAs of Japanese encephalitis virus in C6/36 mosquito cells. A persistent infection was established by replacing growth medium on surviving cells and continued cell passaging. Persistent infection was demonstrated by a continual release of infectious virus, fluorescent antibody staining, and Northern analysis. A population of DI RNAs of approximately 8.2-9.7 kb, not detectable in acutely infected cells, became apparent in the persistently infected cells by 25 days postinfection. Sequence analyses revealed a population of DI RNAs that contained in-frame deletions of 1.3-2.8 kb covering the region of the E gene and some flanking C or prM and NS1 gene sequences. Transcripts from one cDNA clone of a DI RNA replicated in uninfected mosquito cells as demonstrated by RT-PCR. DI RNA-containing virions in supernatant fluids from persistently infected mosquito cells could be used to establish persistent infection in BHK-21 cells. The correlation of DI RNA presence with cell survival suggests that DI RNAs are contributing mechanistically to the establishment of persistent infection in both the mosquito and mammalian cells. PMID- 17134785 TI - [Training programs in stroke patients: literature review]. AB - INTRODUCTION: An increased number of physical retraining programs are available for the rehabilitation of stroke patient; generally positive results are observed, but no consensus in methodology exists (program characteristics, intensity, duration, frequency, etc.). OBJECTIVE: To review the literature about training program characteristics for stroke patients and describe their efficacy, limits, and results. METHODS: A search of the Medline database revealed 97 references, including 53 clinical studies examining protocols of physical training and 15 meta-analyses and literature reviews of physical training. RESULTS: Three physical training protocols were found (walking, neuromuscular and 'force' training). Each training had specific effects: cardiovascular, functional, and muscular. DISCUSSION-CONCLUSION: To obtain positive cardiovascular effects and/or walking improvement, classical rehabilitation and physical training programs need to be associated for rehabilitation of the stroke patient. PMID- 17134786 TI - Sustained beneficial metabolic effects 18 months after a 30-day very low calorie diet in severely obese, insulin-treated patients with type 2 diabetes. AB - Eighteen insulin-treated obese type 2 diabetic patients were followed for 18 months after they followed a 30-day very low calorie diet (VLCD, 450kCal/day) with the cessation of all glucose-lowering medication. After the 30-day VLCD, caloric intake was slowly increased to eucaloric and glucose-lowering medication was restarted if necessary. On day 0 and 30 of the VLCD and after 18 months follow-up, bodyweight, blood-pressure, glycaemic control and lipid levels were measured. The 30-day VLCD significantly reduced bodyweight (-11.7+/-0.7kg, mean+/ S.E.M.) and improved dyslipidaemia, hypertension and glycaemia. As a group, this effect was sustained at 18 months follow-up despite the fact that patients used less lipid-, blood-pressure- and glucose-lowering medication. Especially, the use of insulin was significantly reduced: 18 out of 18 patients on day 0 (mean 137+/ 22units/day); 5 out of 18 patients at 18 months (86+/-14units/day). Patients using insulin at 18 months had regained weight to prediet levels, but still had a better cardiovascular risk profile compared with before the dietary intervention. Thus, a once-only 30-day VLCD leads to a sustained improvement in glycaemia, dyslipidaemia and blood-pressure up to 18 months follow-up in obese type 2 diabetic patients, even, although to a lesser extent, in patients who regained body-weight. PMID- 17134787 TI - The impact of cost sharing of prescription drug expenditures on health care utilization by the elderly: own- and cross-price elasticities. AB - PURPOSE: To estimate healthcare demand elasticity and evaluate the impact of deductible/co-payment policy changes for prescription drugs on the use of drugs and physician visits among seniors with rheumatoid arthritis (RA) in British Columbia (BC), Canada. METHODS: According to the BC drug insurance program, prior to 2002, seniors co-paid the dispensing fee of each prescription to an annual maximum of CAN$ 200 (plan A). Starting in 2002, this plan was split into plan A and plan A1 (Premium Assistance) such that the co-payment amount equaled a maximum of CAN$ 25 and CAN$ 10 per prescription to an annual maximum of CAN$ 275 and CAN$ 200, respectively. Because of the endogeneity of the beneficiary price in the presence of a non-linear price schedule resulting from the cost-sharing policy, we implemented the method of instrumental variables to estimate price elasticities. The instrument was based on the price an individual would face under the new cost-sharing policy if their consumption remained at the pre-policy level. RESULTS: A total of 8017 patients were included. The estimated own-price elasticity of demand for prescription drugs and the cross-price elasticity of demand for physician visits were found to be negative and positive, respectively. The implications of our findings were that when cost sharing for prescription drugs increased, the demand for prescription drugs decreased and the demand for physician visits increased. CONCLUSIONS: In a predominantly publicly funded health care system, the selective introduction of market driven cost containment concepts such as patient cost-sharing might have the unintended impact of increasing overall health utilization for seniors with RA. PMID- 17134788 TI - Adenovirus expressing shRNA to IGF-1R enhances the chemosensitivity of lung cancer cell lines by blocking IGF-1 pathway. AB - RNA interference is a phenomenon whereby small double-stranded RNA knocks down the expression of a sequence-specific gene. Double-stranded siRNA transfection, as currently used, is considered to have transient and low transfection efficiency. We constructed an adenoviral vector-based short hair-pin(sh)RNA system to overcome the limitations of the genetic blockade of IGF-1R, one of most important cancer therapy targets. We constructed three different IGF-1R specific shRNAs (612, 801, and 3425) and generated three ad-shIGF-1Rs using BD Adeno-X expression system. We assessed the effect of ad-shIGF-1R on signal transduction, induction of apoptosis, and in vitro tumorigenicity of lung cancer cell lines. Western blot and FACS assays demonstrated that endogenous IGF-1R expression was efficiently suppressed after transduction of lung cancer cell lines with the three different ad-shIGF-1Rs. IGF-1R blockade by ad-shIGF-1R inhibited ligand induced phosphorylation of pAkt and pErk, and ad-shIGF-1R effectively blocked the in vitro tumorigenicity of lung cancer cell lines. Moreover, the transduction of a human lung cancer cell line with ad-IGF-1R(3425) enhanced chemosensitivity to anticancer drugs. We conclude that the adenoviral vector-based approach to the RNA interference of IGF-1R induced effective IGF-1R silencing in lung cancer cell lines as manifested by effective blocking of the downstream pathway of IGF-1R and by an antitumor effect. We believe that this system can be usefully applied to other cancer targets. PMID- 17134789 TI - Evaluation of extratumoral lymphatic permeation in non-small cell lung cancer as a means of predicting outcome. AB - BACKGROUND: Lymphatic permeation (ly) has been described as a potential prognostic factor for non-small cell lung cancer (NSCLC). METHODS: The purpose of this study was to analyze whether evaluation of the presence or absence of ly and its location (ly 0: absent, N=464; ly 1: intratumoral, N=42; ly 2: extratumoral, N=52) provides an appropriate means of predicting the outcome of NSCLC. We investigated the clinical implications of ly in 558 consecutive patients with surgically resected NSCLC. RESULTS: Evaluation according to ly status showed that the recurrence-free survival (RFS) time of the ly 2 patients was significantly shorter than that of the ly 0 patients (P<0.0001), the ly 1 patients (P=0.0028). A significant difference in RFS time was also observed between the ly 0 patients and the ly 1 patients (P=0.0025). RFS time of the ly 0 patients was significantly longer than that of the ly 1 plus ly 2 patients (P<0.0001). We also evaluated the patients with pathological stage I disease (N=378) separately. The RFS time of the ly 2 patients (N=9) was significantly shorter than that of the ly 0 plus ly 1 patients (P<0.0001). In the nine ly 2 patients, six developed a distant metastasis within 1 year. A multivariate analysis revealed that ly status (ly 0 plus ly 1 versus ly 2) was an independent prognostic factor (P=0.0116), demonstrating the significant prognostic value of extratumoral lymphatic permeation in NSCLC. CONCLUSIONS: These results indicate that ly status is a good prognostic marker of poorer outcome in patients with resected NSCLC. PMID- 17134790 TI - The chemical response of particle-associated contaminants in aquatic sediments to urbanization in New England, U.S.A. AB - Relations between urbanization and particle-associated contaminants in New England were evaluated using a combination of samples from sediment cores, streambed sediments, and suspended stream sediments. Concentrations of PAHs, PCBs, DDT, and seven trace metals (Cd, Cr, Cu, Hg, Ni, Pb, Zn) were correlated strongly with urbanization, with the strongest relations to percentage commercial, industrial, and transportation (CIT) land use. Average PAH and metal concentrations in the most urbanized watersheds were approximately 30 and 6 times the reference concentrations, respectively, in remote, undeveloped watersheds. One-quarter to one-half of sampling sites had concentrations of PAHs, Cu, Pb, or Zn above the probable effects concentration, a set of sediment quality guidelines for adverse effects to aquatic biota, and sediments were predicted to be toxic, on average, when CIT land use exceeded about 10%. Trends in metals in cores from urban watersheds were dominantly downward, whereas trends in PAHs in a suburban watershed were upward. A regional atmospheric-fallout gradient was indicated by as much as order-of-magnitude-greater concentrations and accumulation rates of contaminants in cores from an undeveloped reference lake in Boston compared to those from remote reference watersheds. Contaminant accumulation rates in the lakes with urbanization in their watersheds, however, were 1-3 orders of magnitude greater than those of reference lakes, which indicate the dominance of local sources and fluvial transport of contaminants to urban lakes. These analyses demonstrate the magnitude of urban contamination of aquatic systems and air sheds, and suggest that, despite reductions in contaminant emissions in urban settings, streams and lakes will decline in quality as urbanization of their watersheds takes place. PMID- 17134791 TI - Comparison of the solution behaviour of a pyrite-calcite mixture in batch and unsaturated sand column. AB - A successful application of reaction transport algorithms to calculate the chemical evolution of natural systems requires accurate methods to compute the rates of mineral/fluid surface reactions. Regarding the transport of radio nuclides in mining dumps the dissolution of minerals is of special importance. Using a kinetic rate law of the mineral dissolution verified for unsaturated conditions will allow a realistic modelling of the mineral weathering in the environment. Dissolution rates of minerals in an aqueous solution are determined by several characteristics. These are surface reaction rates, morphology of the mineral's surface and, in case it is the unsaturated zone, the degree of the water saturation. For this process, the quantity of the particle surfaces which are in contact with percolating water is most decisive. In order to study the differences of mineral dissolution under saturated and unsaturated conditions batch and column experiments were carried out with a pyrite-calcite mixture. The experimental results were verified by calculations. Comparing the dissolution in batch with those in the column experiment, which was performed with a water flow velocity of 0.64 cm/day and was analyzed in the region of a water saturation of 0.11, one can conclude that only a small portion of about 5% of the grain surface is chemically reactive in this unsaturated flow. PMID- 17134792 TI - A C-terminal aldehyde analog of the insect kinins inhibits diuresis in the housefly. AB - The insect kinins are present in a wide variety of insects and function as potent diuretic peptides in flies. A C-terminal aldehyde insect kinin analog, Fmoc RFFPWG-H (R-LK-CHO), demonstrates stimulation of Malpighian tubule fluid secretion in crickets, but shows inhibition of both in vitro and in vivo diuresis in the housefly. R-LK-CHO reduced the total amount of urine voided over 3 h from flies injected with 1 microL of distilled water by almost 50%. The analog not only inhibits stimulation of housefly fluid secretion by the native kinin Musdo K, but also by thapsigargin, a SERCA inhibitor, and by ionomycin, a calcium ionophore. The activity of R-LK-CHO is selective, however, as related C-terminal aldehyde analogs do not demonstrate an inhibitory response on housefly fluid secretion. The selective inhibitory activity of R-LK-CHO on housefly tubules represents an important lead in the development of environmentally friendly insect management agents based on the insect kinins. PMID- 17134793 TI - What is the impact of theoretical knowledge on children's nurses' post-operative pain management practices? An exploratory study. AB - Despite the availability of the evidence to guide pain management practices, practices are often sub-optimal with children experiencing moderate to severe pain post-operatively. Limited theoretical knowledge about managing pain has been suggested as one reason for this. Several studies have identified gaps in nurses' theoretical knowledge. However, the affect of theoretical knowledge on pain management practices has not been explored. This explored whether there is a relationship between nurses' theoretical knowledge and the quality of their practices. Nurses (n=13) on one children's surgical ward were shadowed for a five hour period during two-four shifts. Data about post-operative pain management practices were collected using a pain management checklist and field notes. Nurses (n=12) also completed the revised pain management knowledge test. Questionnaire scores were compared to the observational data. No positive relationship was found between nurses' level of theoretical knowledge and how well they actually managed pain. Nurses did not appear to routinely apply theoretical knowledge in practice. This may explain, at least in part, why pain management practices remain poor despite the evidence to guide practice being readily available. The hypothesis, put forward in other studies, that increasing nurses' theoretical knowledge about pain will improve practices may be overly simplistic. PMID- 17134794 TI - Developing professional habitus: a Bernsteinian analysis of the modern nurse apprenticeship. AB - This paper discusses the development of professional habitus in nursing students from a sociological, and specifically, a Bernsteinian perspective. It outlines the theoretical basis for the development of this trait, regarded as one of the defining characteristics of professional practice, and discusses how its development has shaped the modern nurse apprenticeship with its current emphasis on situated learning. The paper considers some of the pitfalls to this approach and raises some caveats about the assumptions which underpin nursing education at the current time. It discusses how students' legitimate peripheral participation in the workplace may be jeopardised, and outlines how they may be faced with untenable choices in respect of 'fitting in' to the ward team or challenging poor practice where this occurs. Moreover, the paper considers how the increasing abrogation of 'caring' activities to non-registered staff threatens the very notion of professional habitus in nursing and posits some possible explanations for this. The paper concludes by arguing that a better understanding of professional habitus is required by all within the profession, and suggests that this concept provides the means by which two seemingly disparate concepts, 'professionalism' and 'vocationalism' can be brought together to the benefit of the nursing profession. PMID- 17134795 TI - Long-term humoral and cell-mediated immunity after acellular pertussis vaccination compares favourably with whole-cell vaccines 6 years after booster vaccination in the second year of life. AB - Humoral and cell-mediated immune responses (CMI) were evaluated in subjects 3 and 6 years after primary and booster vaccination with either three-component acellular (Pa) or whole-cell (Pw) vaccines. Low anti-pertussis toxin (PT) antibody levels confirmed the absence of pertussis disease, consistent with ongoing protection. Anti-pertactin (PRN) antibodies, remained at higher levels in Pa-vaccinated subjects. At year 6, CMI responses continued to be present and were higher in Pa-vaccinated than Pw-vaccinated subjects. Long-term protection with Pa vaccines can be expected to be at least as good as that provided by efficacious Pw vaccines. PMID- 17134796 TI - Mallet fractures: a novel approach to internal fixation using a hook plate. AB - A new treatment of mallet fractures of the distal phalanx is presented. Open reduction and internal fixation was performed using a "hook" plate fabricated from a 1.3 mm AO hand modular system straight plate. This technique avoids the need to place implants or wires through the small avulsion fragment while still being able to achieve a stable "tension-plate" type of fixation construct strong enough to allow protected early active motion of the distal interphalangeal joint. In minimising the need for prolonged splinting, patient comfort is also improved. In a consecutive series of nine fractures, union was achieved in all cases. At an average follow-up period of 17 months, four had excellent and five had good results using the Crawford rating scale. Using the Warren and Norris scale, all patients had a successful result. The final average active range of flexion of the distal interphalangeal joint was 64 degrees and there was no extensor lag. PMID- 17134797 TI - Enhanced sensory recovery after median nerve repair using cortical audio-tactile interaction. A randomised multicentre study. AB - The "Sensor Glove System" offers an alternate afferent inflow from the hand early after nerve repair in the forearm, mediated through the hearing sense, implying that deprivation of one sense can be compensated by another sense. This sensory "by-pass" was used early after repair of the median nerve with the intention of improving recovery of functional sensibility by maintaining an active sensory map of the hand in the somatosensory cortex during the deafferentation period. In a prospective multicentre clinical study, one group (n=14) started early after surgery with sensory re-education using the Sensor Glove System and the control group (n=12) received conventional sensory re-education, starting 3 months postoperatively. The patients were checked regularly during a 1-year period, with focus on recovery of tactile gnosis. After 12, months, tactile gnosis was significantly better in the Sensor Glove System group. This highlights the timing for introduction of training after nerve repair, focusing on the importance of immediate sensory re-learning. PMID- 17134798 TI - The use of a proximally based posterior interosseous adipofascial flap to prevent recurrence of synostosis of the elbow joint and forearm. AB - We report on four children in whom a proximally based, posterior interosseous artery adipofascial flap was used as an adjunct to surgical resection of synostoses of the forearm and elbow. Three traumatic radio-ulnar and one congenital humero-radial synostoses were treated. The postoperative pronation to supination arc of motion was excellent in all of the traumatic cases and fair in the congenital case. PMID- 17134799 TI - Presentation of varicella-zoster mimicking carpal tunnel syndrome. PMID- 17134800 TI - Locked vs. unlocked plate osteosynthesis of the proximal humerus - a biomechanical study. AB - BACKGROUND: Locked plates (internal fixators) have been found to be an optimal method for the fixation in proximal humeral fractures. In a biomechanical cadaver study the difference between locked and non-locked osteosyntheses was investigated. METHODS: Paired humeri were harvested, bone density measured. Locked internal fixators were mounted on one specimen; identical plate-screw systems without locking mechanism applied to the contralateral specimen for comparison. After that, a transverse subcapital osteotomy was performed. With 7 pairs of humeri static tests with increasing axial loads and with 5 pairs dynamic tests with 10 N preload and 80 N maximal axial load for up to 1 million cycles were performed. RESULTS: In the static experiments the elastic stiffness of the construct was 74% higher in the locked group (median 80 N/mm, quartile range 77 86 N/mm) compared with the non-locked group (46 N/mm, 35.5-56.5 N/mm). The difference was statistically significant (Wilcoxon test for paired samples, P<0.05). Similarly, the linear range until failure was definitely extended in the locked group by 64% (92 N, 89-98 N vs. 56 N, 36.5-73.5 N, P<0.05). Under dynamic loading the non-locked group showed fixation failures between 97,000 and 500,000 cycles. In the locked group no failure was observed until the end of the experiment at 1 million cycles (P<0.0.5). The final deformation was found to be 1 mm (median, quartil range 1.0-1.2 mm) in the non-locked group and 0.3 mm (0.2-0.3 mm) in the locked group (P<0.05). The differences were found equally in lower as well as in higher bone density specimen. INTERPRETATION: Because of the optimal load transfer between implant and cancellous bone, a locked screw plate interface will reduce fixation failure in proximal humeral fractures. PMID- 17134801 TI - Morphological muscle and joint parameters for musculoskeletal modelling of the lower extremity. AB - BACKGROUND: To assist in the treatment of gait disorders, an inverse and forward 3D musculoskeletal model of the lower extremity will be useful that allows to evaluate if-then scenarios. Currently available anatomical datasets do not comprise sufficiently accurate and complete information to construct such a model. The aim of this paper is to present a complete and consistent anatomical dataset, containing the orientations of joints (hip, knee, ankle and subtalar joints), muscle parameters (optimum length, physiological cross sectional area), and geometrical parameters (attachment sites, 'via' points). METHODS: One lower extremity, taken from a male embalmed specimen, was studied. Position and geometry were measured with a 3D-digitizer. Optotrak was used for measurement of rotation axes of joints. Sarcomere length was measured by laser diffraction. FINDINGS: A total of 38 muscles were measured. Each muscle was divided in different muscle lines of action based on muscle morphology. 14 Ligaments of the hip, knee and ankle were included. INTERPRETATION: The presented anatomical dataset embraces all necessary data for state of the art musculoskeletal modelling of the lower extremity. Implementation of these data into an (existing) model is likely to significantly improve the estimation of muscle forces and will thus make the use of the model as a clinical tool more feasible. PMID- 17134802 TI - A method for patient-specific evaluation of vertebral cancellous bone strength: in vitro validation. AB - BACKGROUND: In the context of osteoporosis, important determinants of the fracture risk are the apparent strength and stiffness of cancellous bone, as well as its brittleness and energy absorption capacity. Standard medical imaging, however, cannot measure these mechanical properties directly. Consequently, an estimation of the risk for fracture is made by correlating relative density or mineral density at a skeletal site with statistics of fracture occurrence, which provides limited and partial indications on fracture risks. A better method for evaluating the patient-specific mechanical properties of cancellous bone is therefore required. METHODS: In order to asses the mechanical properties of vertebral cancellous bone, we developed a finite element parametric model of lattice trabecular architecture that, in the future, will be suitable for use with bone imaging modalities. The model inputs are apparent morphological parameters (trabecular thickness and trabecular separation) and the bone mineral density. We conducted uniaxial compression tests on 36 canine vertebral cancellous bone specimens (C7 and L1) to validate model predictions of strength and stiffness in vitro. FINDINGS: Predictions of strength and stiffness matched the experimental results within relative absolute errors of 17.7% and 12.8%, respectively (average of differences between model-predicted and measured values, divided by the average of measured values). We also employed the model for evaluation of strength and stiffness of human L1 and L5 vertebrae and found mean strength of 1.67 MPa (confidence interval 0.42 MPa) and mean elastic modulus of 190 MPa (confidence interval 50 MPa), which are well within the range of previously reported apparent strength and stiffness properties. INTERPRETATION: The present model can be used to improve medical imaging-based evaluation of the spine in osteoporotic individuals by providing more specific information on the individual bone's susceptibility to fracture once clinical bone scans will be able to provide more reliable measures of trabecular thickness and separation. PMID- 17134803 TI - Linking valve closure behavior and sodium transport mechanism in freshwater clam Corbicula fluminea in response to copper. AB - The purpose of this study is to develop a mechanistic model to describe a conceptually new "flux-biological response" approach based on biotic ligand model (BLM) and Michaelis-Menten (M-M) kinetics to allow the linkage between valve closure behavior and sodium (Na) transport mechanism in freshwater clam Corbicula fluminea in response to waterborne copper (Cu). We test the proposed model against published data regarding Na uptake kinetics in rainbow trout and Na uptake profile in C. fluminea, confirming that the predictive model is robust. Here, we show that the predicted M-M maximum Cu internalization flux in C. fluminea is 0.369 micromolg(-1)h(-1) with a half-saturation affinity constant of 7.87x10(-3) microM. Dynamics of Na uptake and valve closure daily rhythm driven by external Cu can also be predicted simultaneously. We suggest that this "Na transport-valve closure behavior" approach might provide the basis of a future design of biomonitoring tool. PMID- 17134804 TI - Influence of hydrological regime on pore water metal concentrations in a contaminated sediment-derived soil. AB - Options for wetland creation or restoration might be limited because of the presence of contaminants in the soil. The influence of hydrological management on the pore water concentrations of Cd, Cr, Cu, Fe, Mn, Ni and Zn in the upper soil layer of a contaminated overbank sedimentation zone was investigated in a greenhouse experiment. Flooding conditions led to increased Fe, Mn, Ni and Cr concentrations and decreased Cd, Cu and Zn concentrations in the pore water of the upper soil layer. Keeping the soil at field capacity resulted in a low pore water concentration of Fe, Mn and Ni while the Cd, Cu, Cr and Zn concentrations increased. Alternating hydrological conditions caused metal concentrations in the pore water to fluctuate. Formation and re-oxidation of small amounts of sulphides appeared dominant in determining the mobility of Cd, Cu, and to a lesser extent Zn, while Ni behaviour was consistent with Fe/Mn oxidation and reduction. These effects were strongly dependent on the duration of the flooded periods. The shorter the flooded periods, the better the metal concentrations could be linked to the mobility of Ca in the pore water, which is attributed to a fluctuating CO(2) pressure. PMID- 17134805 TI - Multivariate statistical evaluation of trace elements in groundwater in a coastal area in Shenzhen, China. AB - Multivariate statistical techniques are efficient ways to display complex relationships among many objects. An attempt was made to study the data of trace elements in groundwater using multivariate statistical techniques such as principal component analysis (PCA), Q-mode factor analysis and cluster analysis. The original matrix consisted of 17 trace elements estimated from 55 groundwater samples colleted in 27 wells located in a coastal area in Shenzhen, China. PCA results show that trace elements of V, Cr, As, Mo, W, and U with greatest positive loadings typically occur as soluble oxyanions in oxidizing waters, while Mn and Co with greatest negative loadings are generally more soluble within oxygen depleted groundwater. Cluster analyses demonstrate that most groundwater samples collected from the same well in the study area during summer and winter still fall into the same group. This study also demonstrates the usefulness of multivariate statistical analysis in hydrochemical studies. PMID- 17134806 TI - Comparing sediment quality in Spanish littoral areas affected by acute (Prestige, 2002) and chronic (Bay of Algeciras) oil spills. AB - The quality of sediments collected from two areas of the Spanish coast affected by different sources of contaminants has been compared in this study. The areas studied are the coast of Galicia affected by the oil spill from the tanker Prestige (November 2002) and the Gulf of Cadiz which suffers continuous inputs of contaminants from industries located in the area and from oil spills. Contamination by several chemicals (metals, PCBs and PAHs) that bind to sediments was analyzed, and two toxicity tests (Microtox) and amphipod 10-day bioassay) were conducted. PAHs were identified as the compounds responsible for the toxic effects. Results show differences between an acute impact related to the sinking of the tanker Prestige and the chronic impact associated with continuous oil spills associated with the maritime and industrial activities in the Bay of Algeciras, this being the most polluted part of the two coastal areas studied in this work. PMID- 17134807 TI - Occurrence of alkylphenolic substances in a Great Lakes coastal marsh, Cootes Paradise, ON, Canada. AB - Occurrence and fate of alkylphenols (APs), known endocrine disruptors, were investigated in a Great Lakes coastal wetland, Cootes Paradise, ON. The wetland, which receives discharges from a Wastewater Treatment Plant (WTP) and several Combined Sewer Overflows (CSOs), is an important spawning ground for fish and crucial habitat for other fauna. Elevated concentrations of nonylphenol ethoxylates (NPEs) and their degradation product nonylphenol (NP) were found in water and sediment samples near the sources. Since transfer of APs through the food chain is of concern, we compared their concentrations in invertebrates from clean and contaminated sites. The results reveal transfer of alkylphenolics from sediments to biota and their accumulation in the invertebrate tissue, particularly the highly hydrophobic 4-NP, whose concentrations ranged from 1.9 to 6.3 microg g(-1). To our knowledge, this is the first study to evaluate AP concentrations in tissue of benthic invertebrates under real environmental conditions. PMID- 17134808 TI - Development and validation of a terrestrial biotic ligand model predicting the effect of cobalt on root growth of barley (Hordeum vulgare). AB - A Biotic Ligand Model was developed predicting the effect of cobalt on root growth of barley (Hordeum vulgare) in nutrient solutions. The extent to which Ca(2+), Mg(2+), Na(+), K(+) ions and pH independently affect cobalt toxicity to barley was studied. With increasing activities of Mg(2+), and to a lesser extent also K(+), the 4-d EC50(Co2+) increased linearly, while Ca(2+), Na(+) and H(+) activities did not affect Co(2+) toxicity. Stability constants for the binding of Co(2+), Mg(2+) and K(+) to the biotic ligand were obtained: logK(CoBL)=5.14, logK(MgBL)=3.86 and logK(KBL)=2.50. Limited validation of the model with one standard artificial soil and one standard field soil showed that the 4-d EC50(Co2+) could only be predicted within a factor of four from the observed values, indicating further refinement of the BLM is needed. PMID- 17134809 TI - Risk assessment of lutein and lycopene. PMID- 17134810 TI - Comments on "Low plasma BDNF is associated with suicidal behavior in major depression" by Y.K. Kim et al. Progress in Neuropsychopharmacology and Biological Psychiatry 2006 Aug 9; [Epub ahead of print]. PMID- 17134811 TI - Embryo-fetal development toxicity of prenatal exposure to penequine hydrochloride intramuscularly in rats. AB - The potential for penequine hydrochloride to induce maternal and embryo-fetal developmental toxicity was evaluated in Wistar rats. The drug was administered intramuscularly (i.m.) at dose levels of 0, 10, 30 or 50mg/kg/day to groups of pregnant rats from day 6 to 15 of gestation. All dams were observed for maternal body weights, food consumption and any abnormal change, and subjected to caesarean-section on gestation day (GD) 20; all fetuses obtained from caesarean section were assessed by external inspection, visceral and skeletal examinations. In the 50mg/kg/day group, maternal toxicity included an increase in the incidence of abnormal clinical signs, and decrease in the body weight and body weight gain. Developmental toxicity included an increase in the postimplantation loss, a decrease in the litter size, and a reduction in the gravid uterus weight. In addition, a statistically non-significant increase in the incidence of fetal external, visceral, and skeletal alterations including malformations and variations were seen in high-dose group. There were no treatment-related findings in maternal clinical and intrauterine observations, and fetal morphological examinations in mid-, low-dose and control groups. Thus, under the conditions of this study, the no-observed-adverse-effect-level (NOAEL) and lowest-observed adverse-effect-level (LOAEL) of penequine hydrochloride for both maternal and embryo-fetal toxicity in the Wistar rats were considered to be 30mg/kg/day and 50mg/kg/day, which are approximately 900 and 1500 above the therapeutic dosage, respectively. PMID- 17134812 TI - [The chondrosarcoma of the hand: review of the literature about a case and a proposition of a decision-making therapeutic tree]. AB - We report a case of chondrosarcoma of the hand initially treated as a chondroma. Through a review of the literature, we review the clinical, radiological and histological diagnostic difficulties of this lesion. We propose a decision-making tree for the surgical coverage of the chondrosarcoma of the hand. PMID- 17134813 TI - Matrine induced gastric cancer MKN45 cells apoptosis via increasing pro-apoptotic molecules of Bcl-2 family. AB - Matrine, one of the main active components from the dry roots of Sophora flavescence, was known to induce apoptosis in a variety of tumor cells in vitro. However, the molecular mechanism of cell apoptosis induced by Matrine remains elusive. Here, we investigated the apoptosis in Matrine-treated human gastric cancer MKN45 cells. The results showed that Matrine could inhibit cell proliferation and induce apoptosis in a dose-dependent manner. Further immunoblots revealed that in Matrine-treated cells, caspase-3, -7 were activated and the pro-apoptotic molecules Bok, Bak, Bax, Puma, and Bim were also up regulated. Our results suggested that Matrine induced gastric cancer MKN45 cells apoptosis via increasing pro-apoptotic molecules of Bcl-2 family. PMID- 17134814 TI - The 'ABC for life' programme - teaching basic life support in schools. AB - BACKGROUND: Less than 1% of the general public know how to assess or manage someone who has collapsed. It has been estimated that if 15-20% of the population were capable of performing cardiopulmonary resuscitation (CPR), mortality of out of hospital cardiac arrest could be decreased significantly. Training basic life support (BLS) skills to school children would be the most cost effective way of achieving this goal and ensuring that a large proportion of the population acquire basic life saving skills. AIMS: To assess retention of knowledge of basic life support 6 months after a single course of instruction in cardiopulmonary resuscitation designed specifically for school children. SETTING: School pupils in a rural location in one region of the United Kingdom. METHODS: A course of instruction in cardiopulmonary resuscitation - the 'ABC for life' programme - specifically designed to teach 10-12-year-old school children basic life support skills. The training session was given to school pupils in a rural location in Northern Ireland. A 22 point questionnaire was used to assess acquisition and retention of basic life support knowledge. RESULTS: Children instructed in cardiopulmonary resuscitation showed a highly significant increase in level of knowledge following the training session. While their level of knowledge decreased over a period of 6 months it remained significantly higher than that of a comparable group of children who had never been trained. CONCLUSION: A training programme designed and taught as part of the school curriculum would have a significant impact on public health. PMID- 17134815 TI - A comparison of electrically induced cardiac arrest with cardiac arrest produced by coronary occlusion. AB - OBJECTIVE: The present study was undertaken to compare an animal model of electrically induced VF with ischemically induced VF. In a preponderance of models of cardiac arrest and resuscitation in intact animals, ventricular fibrillation (VF) is induced by an alternating current delivered directly to the epicardium or endocardium. Yet, the applicability of such animal models has been challenged for it is not an electrical current alone but rather a current generated in the ischemic myocardium that triggers VF. Accordingly, a potentially more clinically relevant model was investigated in which spontaneous VF followed acute myocardial ischemia. METHODS: Twenty anesthetized pigs were randomized to either electrical fibrillation or myocardial ischemia following transient occlusion of the left anterior descending (LAD) coronary artery. RESULTS: VF was untreated for 7 min in both models after which mechanical ventilation and precordial compression were begun. Defibrillation was attempted after 5 min of CPR in both groups. VF appeared within 5.7+/-2.0 min of LAD occlusion. CONCLUSIONS: A significant increase in the number of post-resuscitation premature ventricular beats and recurrent VF followed ROSC and a significantly greater number of shocks was required for restoration of spontaneous circulation (ROSC) after LAD occlusion. Nevertheless, early post-resuscitation myocardial dysfunction, neurological recovery and 72 h survival were indistinguishable between the two models. PMID- 17134816 TI - Late-onset and recurrent neonatal Streptococcus agalactiae infection with ingestion of infected mother's milk. PMID- 17134817 TI - Occurrence of ST-changes recorded with the STAN S21-monitor during normal and abnormal fetal heart rate patterns during labour. AB - OBJECTIVE: To determine the frequency of ST-changes in the fetal electrocardiogram (ECG) both during normal and abnormal fetal heart (FHR) traces and to study relations with characteristics of pregnancy and delivery. STUDY DESIGN: Retrospective analysis of 563 FHR+ECG-recordings. ST-changes were counted for all recordings with respect to their FHR pattern. Relationships between the occurrence of ST-events and characteristics of pregnancy and delivery were determined for the recordings with a completely normal FHR tracing during the first stage. RESULTS: ST-events were present during 51.7% of recordings in the first stage of labour and during 24.3% of the recordings in the second stage. Surprisingly, events occurred during the first stage at a similar incidence during normal and abnormal FHR-patterns. During the second stage of labour events also occurred in a similar percentage of normal, intermediary and abnormal recordings, but - if present - events were more numerous during the abnormal FHR traces. In the subgroup of cases with a normal FHR pattern more events per hour occurred in babies born at 36-37 weeks of gestation and in boys. CONCLUSION: ST events are a frequent finding during normal FHR tracings. PMID- 17134818 TI - Blue peritoneal implants--is it always endometriosis? PMID- 17134819 TI - Thermal denaturations of staphylococcal nuclease wild-type and mutants monitored by fluorescence and circular dichroism are similar: lack of evidence for other than a two state thermal denaturation. AB - It is unclear whether the thermal denaturation of staphylococcal nuclease is a two state, three state, or variable two state process. The thermal denaturation of wild-type staphylococcal nuclease was followed by tryptophan fluorescence and circular dichroism signal at 222 nm, forty-two and fourteen times, respectively. Analysis of this data using a simple two state model gave melting temperatures of 53.0+/-0.4 degrees C (fluorescence) and 52.7+/-0.6 degrees C (CD) and van't Hoff enthalpies of 82.4+/-2.6 kcal/mol and 88.6+/-4.2 kcal/mol. Ninety-seven mutants also had these parameters determined by both fluorescence and CD. The average difference between the melting temperatures was 1.05+/-0.75 degrees and the average difference between van't Hoff enthalpies was 1.6+/-4.8 kcal/mol. These very similar results for the two spectroscopic probes of structure are discussed in the context of the different models that have been proposed for nuclease denaturation. It is concluded, for most nuclease variants, that the errors introduced by a two state assumption are negligible and either virtually all helical structure is lost in any initial unfolding event or any intermediate must have low stability. PMID- 17134820 TI - Preliminary evaluation of the electrochemical and chemical coagulation processes in the post-treatment of effluent from an upflow anaerobic sludge blanket (UASB) reactor. AB - The main objective of this paper was to perform a preliminary comparative study between chemical and electrochemical coagulation processes, both followed by flocculation and sedimentation of an effluent from an upflow anaerobic sludge blanket (UASB) reactor treating simulated wastewater from an unbleached Kraft pulp mill. The electrochemical treatment removed up to 67% (with aluminum electrodes) and 82% (with stainless-steel electrodes) of the remaining chemical oxygen demand (COD) and 84% (stainless steel) and 98% (aluminum) of the color in the wastewater. These efficiencies were achieved with an energy consumption ranging from 14 to 20 Wh l(-1). The coagulation-flocculation treatment with ferric chloride and aluminum sulfate removed up to 87% and 90% of COD and 94% and 98% of color, respectively. The addition of a high molecular weight cationic polymer enhanced both COD and color removal efficiencies. The two post-treatment processes proved to be technically feasible; however the economical feasibility could not be assessed since the experiments were performed with small reactors that could distort scale factors. PMID- 17134821 TI - Combining remote sensing imagery and forest age inventory for biomass mapping. AB - Aboveground biomass (AGB) of forests is an important component of the global carbon cycle. In this study, Landsat ETM(+) images and field forest inventory data were used to estimate AGB of forests in Liping County, Guizhou Province, China. Three different vegetation indices, including simple ratio (SR), reduced simple ratio (RSR), and normalized difference vegetation index (NDVI), were calculated from atmospherically corrected ETM(+) reflectance images. A leaf area index (LAI) map was produced from the RSR map using a regression model based on measured LAI and RSR. The LAI map was then used to develop an initial AGB map, from which forest stand age was deduced. Vegetation indices, LAI, and forest stand age were together used to develop AGB estimation models for different forest types through a stepwise regression analysis. Significant predictors of AGB changed with forest types. LAI and NDVI were significant predictors of AGB for Chinese fir (R(2)=0.93). The model using LAI and stand age as predictors explained 94% of the AGB variance for coniferous forests. Stand age captured 79% of the AGB variance for broadleaved forests (R(2)=0.792). AGB of mixed forests was predicted well by LAI and SR (R(2)=0.931). Without differentiating among forest types, the model with SR and LAI as predictors was able to explain 90% of AGB variances of all forests. In Liping County, AGB shows a strong gradient that increases from northeast to southwest. About 64% of the forests have AGB in the range from 90 to 180 t ha(-1). PMID- 17134822 TI - Polycomb-group oncogenes EZH2, BMI1, and RING1 are overexpressed in prostate cancer with adverse pathologic and clinical features. AB - OBJECTIVES: Polycomb group (PcG) proteins are involved in maintenance of cell identity and proliferation. The protein EZH2 is overexpressed in disseminated prostate cancer, implicating a role of PcG complexes in tumor progression. In this study, we evaluated the expression of eight members of both PcG complexes in clinicopathologically defined prostate cancer. METHODS: Components of both PcG protein complexes PRC2 (EZH2, EED, YY1) and PRC1 (BMI1, RING1, HPH1, HPC1, HPC2) were immunohistochemically identified in tissue microarrays of 114 prostate cancer patients. Protein expression was semi-quantitatively scored and correlated with pathologic parameters and recurrence of prostate-specific antigen (PSA). RESULTS: Whereas BMI1, RING1, HPC1 and HPH1 were all abundantly present in normal and malignant prostate epithelium, expression of EZH2 occurred in only <10% of cells. Expression of EZH2, BMI1 and RING1 were all significantly enhanced in tumours with Gleason score (GS) > or = 8, extraprostatic extension, positive surgical margins, and PSA recurrence. When only the subgroup of GS < or = 6 was considered, representing the tumour grade in the majority of needle biopsies, EZH2 and BMI1 were also predictive for PSA recurrence. In a multivariable analysis, BMI1 was the only PcG protein with an independent prognostic value. CONCLUSIONS: PcG proteins EZH2, BMI1, and RING1 are associated with adverse pathologic features and clinical PSA recurrence of prostate cancer. Whereas BMI1 and RING1 are abundantly present in prostate cancer, EZH2 is expressed at relatively low levels, making it a less obvious target for therapy. PMID- 17134823 TI - A semiotic framework for evolutionary and developmental biology. AB - This work aims at constructing a semiotic framework for an expanded evolutionary synthesis grounded on Peirce's universal categories and the six space/time/function relations [Taborsky, E., 2004. The nature of the sign as a WFF--a well-formed formula, SEED J. (Semiosis Evol. Energy Dev.) 4 (4), 5-14] that integrate the Lamarckian (internal/external) and Darwinian (individual/population) cuts. According to these guide lines, it is proposed an attempt to formalize developmental systems theory by using the notion of evolving developing agents (EDA) that provides an internalist model of a general transformative tendency driven by organism's need to cope with environmental uncertainty. Development and evolution are conceived as non-programmed open-ended processes of information increase where EDA reach a functional compromise between: (a) increments of phenotype's uniqueness (stability and specificity) and (b) anticipation to environmental changes. Accordingly, changes in mutual information content between the phenotype/environment drag subsequent changes in mutual information content between genotype/phenotype and genotype/environment at two interwoven scales: individual life cycle (ontogeny) and species time (phylogeny), respectively. Developmental terminal additions along with increment minimization of developmental steps must be positively selected. PMID- 17134824 TI - Establishment and characterization of cell lines from three human thyroid carcinomas: responses to all-trans-retinoic acid and mutations in the BRAF gene. AB - We report the characteristics of three cell lines (designated, SNU-80, SNU-373 and SNU-790), which were established from two papillary carcinomas and one anaplastic carcinoma obtained from three Korean thyroid carcinoma patients. All cell lines grow as adherent cells. Electron microscopy characteristically showed cytoplasmic invaginations of nuclei and intranuclear cytoplasmic inclusions. SNU 80 and SNU-790 cells showed a positive reaction to anti-cytokeratin antibody, and SNU-790 cells positivity for CK-19. All lines were free of mycoplasma or bacteria and were proven unique by DNA fingerprinting analysis. The p15 and p16 genes are deleted in the SNU-790 line. Mutations of the p53 gene were found in two lines (SNU-80 and SNU-373), but no mutations in the RET or MEN1 genes were observed. Mutations of the BRAF gene were found in the SNU-80 (G468R) and the SNU-790 (V599E) cell lines, but no mutations in the K-ras gene were present. SNU-80 and SNU-790 cells showed a positive reaction to anti-cytokeratin antibody, and no evidence of the production of thyroglobulin or calcitonin was observed. The cell lines were unable to trap radioactive iodine but did not contain TSH receptor. In addition, we investigated the mRNA expression levels of Tg, TSHR, TTF-1, PAX-8, NIS, IL-6, and LIF, and of the alpha, beta and gamma retinoic acid receptors in these cell lines. IL-6 was down-regulated in all three cell lines by all-trans retinoic acid treatment. RAR-alpha was expressed but RAR-beta was not expressed in the three cell lines, and RAR-gamma was not expressed in SNU-790. Interestingly, RAR-beta (SNU-80 and SNU-373) and RAR-gamma (SNU-790) was up regulated by all-trans-retinoic acid treatment. We believe that these well characterized thyroid carcinoma cell lines may be useful tools for investigations on the biological characteristics of thyroid carcinoma, particularly for investigations related to gene alterations, especially of the BRAF gene. These cell lines may also be useful for redifferentiation therapy studies on thyroid carcinoma using all-trans-retinoic acid. PMID- 17134825 TI - Direct vaccination with pseudotype baculovirus expressing murine telomerase induces anti-tumor immunity comparable with RNA-electroporated dendritic cells in a murine glioma model. AB - Baculovirus pseudotyped with vesicular stomatitis virus G protein (Bac-VSV-G) was found to efficiently transduce and express transgenes on mammalian cells. In this study, this recombinant virus was used for induction of anti-tumor immunity against murine telomerase reverse transcriptase (mTERT) and was compared with RNA electroporated dendritic cells (DCs) in a murine glioma model. Splenocytes from the mice vaccinated with Bac-VSV-G expressing mTERT (Bac-VSVG-mTERT) showed significantly increased numbers of mTERT-specific IFN-gamma-secreting T cells using an ELISPOT technique, and also showed increased NK cell activity. In addition, the TERT-specific T cells activated by Bac-VSVG-mTERT and mTERT RNA electroporated DCs were predominantly CD4+ T cells and CD8+ T cells, respectively. The protective anti-tumor effect of Bac-VSVG-mTERT was similar to that of mTERT RNA-electroporated DCs. These results suggest that the pseudotype baculovirus expressing TERT may be a good candidate for a genetic vaccine for use in the treatment of malignant gliomas. PMID- 17134826 TI - Spread of large LNG pools on the sea. AB - A review of the standard model of LNG pool spreading on water, comparing it with the model and experiments on oil pool spread from which the LNG model is extrapolated, raises questions about the validity of the former as applied to spills from marine tankers. These questions arise from the difference in fluid density ratios, in the multi-dimensional flow at the pool edge, in the effects of LNG pool boiling at the LNG-water interface, and in the model and experimental initial conditions compared with the inflow conditions from a marine tanker spill. An alternate supercritical flow model is proposed that avoids these difficulties; it predicts significant increase in the maximum pool radius compared with the standard model and is partially corroborated by tests of LNG pool fires on water. Wind driven ocean wave interaction has little effect on either spread model. PMID- 17134827 TI - Degradation and mineralization of Direct Blue 71 in a circulating upflow reactor by UV/TiO2 process and employing a new method in kinetic study. AB - Direct Blue 71 (C(40)H(23)N(7)Na(4)O(13)S(4)), an azo dye with a high worldwide consumption and providing toxic effluents, can be highly degraded using TiO(2) catalyst suspension and irradiation with a UV-C lamp in a circulating upflow photo-reactor with no dead zone. An initial concentration of 50 mgL(-1) of dye, within the range of typical concentration in textile wastewaters, was used. The influence of catalyst concentration, pH and temperature were investigated. The results showed that degradation of this dye can be conducted in the both processes of only UV irradiation and UV/TiO(2); but with the aim of mineralization, the later process provides significantly better results. Accordingly, a degradation of more than 97% of dye was achieved by applying the optimal operational parameters with 40 mgL(-1) of catalyst, natural pH and 45 degrees C, during 120 min irradiation. A removal of about 50% of COD could also be obtained at the same time. In kinetic investigations, the effect of catalyst particles' turbidity was taken into account and the rate of degradation of the dye, under mild conditions, was expressed as the sum of the rates of individual photolysis and photocatalysis process branches, with mainly influence of the bulk hydroxyl radicals. PMID- 17134828 TI - Spatial organization of EEG coherence during listening to consonant and dissonant chords. AB - Theories of harmony state that the contribution of both sensory and cognitive components is important for musical consonance perception. The aims of the present study were to analyze (a) functional intra- and inter-hemispheric connectivity associated with listening to consonant and dissonant chords using EEG coherence method; (b) relationships between affective responsiveness, sensory aspects of perceived consonance and associated brain connectivity. We identified two lines of inter-hemispheric connectivity in the theta band; one localized anterior being sensitive to consonance and one localized posterior sensitive to dissonance. Stronger right intra-hemispheric connectivity for consonance than dissonance in the theta band was associated with higher pleasantness ratings. The relationship between sensory aspects of perceived consonance and left intra hemispheric connectivity found in theta-2 was interpreted as processing of vertical harmony without emotional involvement. The stronger connectivity along the axis "left anterior-right posterior" for dissonance than consonance in the alpha-1 band is discussed as a correlate of novelty processing. By introducing a "auditory object dissociation" hypothesis we suggest to extend the present concept of harmony perception. We believe that "auditory object dissociation" is a component of "sensory dissonance." PMID- 17134829 TI - Regulatory sequences of the PRNP gene influence susceptibility to sporadic Creutzfeldt-Jakob disease. AB - The prion diseases are fatal neurodegenerative disorders that afflict both humans and animals. They comprise kuru, Creutzfeldt-Jakob disease (CJD), Gerstmman Straussler-Scheinker syndrome (GSS), and fatal familial insomnia (FFI). Both GSS, FFI and approximately 10% of CJD cases are genetically linked disorders, whereas 90% of CJD cases are not associated with mutations in the PRNP coding region, therefore other factors must be involved in pathogenesis of these forms of CJD. There is strong evidence that in transgenic mice the level of PrP gene expression influences the initiation and progression of the prion diseases. Moreover, in in vitro experiments demonstrated that mutations in the regulatory region of PRNP gene altered gene expression, therefore it may be expected that PrP expression level influences the susceptibility to CJD. In order to investigate whether single nucleotide polymorphisms within regulatory region of PRNP may modulate genetic susceptibility to sporadic CJD we examined an association of the C/G polymorphism at position -101 with the sCJD. In our study -101G polymorphism is over-represented among sCJD PRNP codon 129M/V cases compared with the control group. Our data suggest that polymorphism at position -101 in the regulatory region of PRNP may be a risk factor for sCJD among codon 129 heterozygotes. PMID- 17134830 TI - Predictors of onset of facial pain and temporomandibular disorders in early adolescence. AB - There are few prospective studies assessing risk factors for onset of temporomandibular (TMD) pain disorders in any age group. The aim of this prospective cohort study was to identify risk factors for onset of clinically significant TMD pain (i.e., pain meeting research diagnostic criteria for myofascial pain and/or arthralgia) during early adolescence. Subjects were 1,996 boys and girls, initially 11 years old, randomly selected from a large nonprofit health care system. Subjects completed a baseline telephone interview and were followed up with mailed questionnaires every 3 months for 3 years. At baseline and all follow ups, subjects were asked to report the presence of facial pain in the past 3 months. Subjects reporting a first onset of facial pain received a standardized clinical examination. In multivariate analyses, baseline predictors of clinically significant pain included female gender [Odds Ratio (OR)=2.0, 95% Confidence Interval (CI)=1.2-3.3] and negative somatic and psychological symptoms including somatization (OR=1.8, CI=1.1-2.8), number of other pain complaints (OR=3.2, CI=1.7-6.1) and life dissatisfaction (OR=4.1, CI=1.9-9.0). Many of the risk factors for onset of clinically significant TMD pain in adolescents are similar to risk factors for onset of TMD and other pain problems in adults, as well as risk factors for onset of other pain conditions in adolescents. These findings suggest that the development of TMD pain in adolescence may reflect an underlying vulnerability to musculoskeletal pain that is not unique to the orofacial region. PMID- 17134831 TI - ASIC3 in muscle mediates mechanical, but not heat, hyperalgesia associated with muscle inflammation. AB - Peripheral initiators of muscle pain are virtually unknown, but likely key to development of chronic pain after muscle insult. The current study tested the hypothesis that ASIC3 in muscle is necessary for development of cutaneous mechanical, but not heat, hyperalgesia induced by muscle inflammation. Using mechanical and heat stimuli, we assessed behavioral responses in ASIC3-/- and ASIC3+/+ mice after induction of carrageenan muscle inflammation. ASIC3-/- mice did not develop cutaneous mechanical hyperalgesia after muscle inflammation when compared to ASIC3+/+ mice; heat hyperalgesia developed similarly between groups. We then tested if the phenotype could be rescued in ASIC3-/- mice by using a recombinant herpes virus vector to express ASIC3 in skin (where testing occurred) or muscle (where inflammation occurred). Infection of mouse DRG neurons with ASIC3-encoding virus resulted in functional expression of ASICs. Injection of ASIC3-encoding virus into muscle or skin of ASIC3-/- mice resulted in ASIC3 mRNA in DRG and protein expression in DRG and the peripheral injection site. Injection of ASIC3-encoding virus into muscle, but not skin, resulted in development of mechanical hyperalgesia similar to that observed in ASIC3+/+ mice. Thus, ASIC3 in primary afferent fibers innervating muscle is critical to development of hyperalgesia that results from muscle insult. PMID- 17134832 TI - The effect of experimenter gender on autonomic and subjective responses to pain stimuli. AB - Several studies have shown that male subjects report lower pain intensity to female compared to male experimenters. The present experiment examined whether experimenter gender also modulated autonomic pain responses. Sixty-four students (32 females) participated in a 2 Subject gender x 2 Experimenter gender x 15 Pain Tests mixed design. Six experimenters, three females and three males collected data. Heat pain was +48 degrees C induced to the right volar forearm. Subjective measurements consisted of pain intensity, pain unpleasantness, stress, arousal and mood. Autonomic responses were heart rate variability and skin conductance levels. The results revealed significant interactions between experimenter gender and subject gender on pain intensity and arousal, but there were no interactions in the physiological data. In conclusion, the lower pain report in male subjects to female experimenters is not mediated by changes in autonomic parameters, and the effect of experimenter gender is probably due to psychosocial factors. PMID- 17134833 TI - Prolonged maintenance of capsaicin-induced hyperalgesia by brief daily vibration stimuli. AB - This study tests the hypothesis that central sensitization initiated by nociceptive input can be maintained by repeated brief innocuous peripheral inputs. Capsaicin was injected intradermally into the hind paw of adult rats. Three different types of daily cutaneous mechanical stimulations (vibration, soft brush, or pressure) were applied to the capsaicin-injected paw for a period of 2 weeks. Daily stimulation consisted of a 10-s stimulation repeated every 30s for 30 min. Foot withdrawal thresholds to von Frey stimuli applied to the paw were measured once a day for 4 weeks. The capsaicin-only group (control rats without daily stimulation) showed hyperalgesia lasting for 3 days. In contrast, hyperalgesia persisted for 2 weeks in the group that received vibration stimulation. Neither the soft brush nor the pressure group showed a significant difference in mechanical threshold from the control group (capsaicin only). The vibration-induced prolonged hyperalgesia was significantly reduced by systemic injection of ifenprodil, an NMDA-receptor antagonist, but it was not influenced by either an AMPA-receptor blocker or a reactive oxygen species (ROS) scavenger. Furthermore, a dorsal column lesion did not interfere with the prolongation of hyperalgesia. Data suggest that vibration-induced prolongation of hyperalgesia is mediated by spinal NMDA-receptors, and a similar mechanism may underlie some forms of chronic pain with no obvious causes, such as complex regional pain syndrome type 1 (CRPS-1). PMID- 17134834 TI - Reduced response to the formalin test and lowered spinal NMDA glutamate receptor binding in adenosine A2A receptor knockout mice. AB - Adenosine is a neuromodulator with complex effects on pain pathways. Mice lacking the adenosine A2A receptor are hypoalgesic, and have altered analgesic responses to receptor-selective opioid agonists. These and other findings suggest a role for the adenosine A2A receptor in sensitizing afferent fibres projecting to the spinal cord. To test this hypothesis formalin (20 microl, 5%) was injected into the paw and nociceptive responses were measured in wildtype and adenosine A2A receptor knockout mice. There was a significant reduction in nociception associated with sensory nerve activation in the knockout mice as measured by time spent biting/licking the formalin-injected paw and number of flinches seen during the first phase, but only the number of flinches was reduced during the second inflammatory phase. In addition, the selective adenosine A2A antagonist SCH58261 (3 and 10 mg/kg) also antagonised both phases of the formalin test. We also labelled NMDA glutamate and NK1 receptors in spinal cord sections as an indirect measure of nociceptive transmission from peripheral sites to the spinal cord. [3H]-Substance P binding to NK1 receptors was unaltered but there was a substantial reduction in binding of [3H]-MK801 to NMDA glutamate receptors in all regions of the spinal cord from knockout mice. The decrease in NMDA glutamate receptor binding may reflect reduced peripheral sensory input to the spinal cord during development and could relate to the hypoalgesia in this genotype. These results support a key role for the adenosine A2A receptor in peripheral nociceptive pathways. PMID- 17134835 TI - Immunohistochemical localization of histamine H3 receptors in rodent skin, dorsal root ganglia, superior cervical ganglia, and spinal cord: potential antinociceptive targets. AB - Activation of histamine H3 receptors (H3Rs) reduces inflammation and nociception, but the existence of H3Rs on peripheral innervation has never been demonstrated. Here we use antibodies to locate H3Rs in whisker pads, hairy and glabrous hind paw skin, dorsal root ganglia (DRGs), and spinal cords of rats, wild type mice, and H3R knockout (H3KO) mice. Although H3Rs have been hypothesized to be on C and sympathetic fibers, H3R-like immunoreactivity (H3R-LI) was only detected on presumptive periarterial A delta fibers and on A beta fibers that terminated in Meissner's corpuscles and as lanceolate endings around hair follicles. The H3R positive periarterial fibers were thin-caliber and coexpressed immunoreactivity for calcitonin gene-related peptide (CGRP), substance P, acid sensing ion channel 3, and 200 kDa neurofilament protein (NF). H3R-LI was also detected on epidermal keratinocytes and Merkel cells, but not on Merkel endings, C fibers, any other A delta fibers, or sympathetic fibers. In DRGs, H3R-LI was preponderantly on medium to large neurons coexpressing NF-LI and mostly CGRP-LI. In dorsal horn, CGRP positive fibers with and without H3R-LI ramified extensively in lamina II; many of the former formed a plexus in lamina V. Low levels of H3R-LI were also present on A beta fibers penetrating superficial and into deeper laminae. The distribution of H3R-LI was similar in rats and wild type mice, but was eliminated or strongly reduced in A delta fibers and A beta fibers, respectively, in H3KO mice. Taken with recently published behavioral results, the present findings suggest that periarterial, peptidergic, H3R-containing A delta fibers may be sources of high threshold mechanical nociception. PMID- 17134836 TI - Observations on the emergence of multiple anthelmintic resistance in sheep flocks in the south-east of Scotland. AB - Multiple resistance to benzimidazole, imidazothiazole and macrocyclic lactone anthelmintics is an emerging problem in the south-east of Scotland. The general management and nematode control strategies employed in four affected flocks (flocks 1-4) were investigated in an attempt to identify the risk factors which might have led to the appearance of production limiting disease associated with anthelmintic resistance. The important risk factors for multiple anthelmintic resistance could not be confirmed and it proved easier to criticise nematode control practices on theoretical grounds, than to propose practical solutions. It seems likely that different risk factors were involved in the four flocks. Lambs in flocks 1 and 2 had been treated with an anthelmintic at 3-4 weekly intervals with the aim of achieving suppressive nematode control, while sheep in flock 1 had been treated with an anthelmintic after they were moved onto clean grazing. Recently lambed ewes had been treated with moxidectin in three of the four flocks, with the aim of controlling their periparturient rise in faecal nematode egg output. All of these factors might have contributed to the emergence of multiple anthelmintic resistance, because they could have led to anthelmintic treatments at times when the nematode population in refugia was small, compared to that in the sheep. Annual rotation of the anthelmintic group was compromised by the emergence of benzimidazole resistance and did not prevent the emergence of multiple resistance in any of the flocks described, although the practice may have slowed the development of resistance. Underdosing may have selected for benzimidazole and imidazothiazole resistance in flock 2, associated with inaccurate estimation of the weights of terminal sire lambs. These investigations also highlighted problems associated with the diagnosis of anthelmintic resistance, in particular the confounding effects of the onset of host immunity to nematode parasites, the possible influence of the age of the adult nematode population, and the insensitivity of the undifferentiated faecal egg count reduction test in situations where resistance is emerging. PMID- 17134837 TI - Molecular characterization of Hepatozoon sp. from Brazilian dogs and its phylogenetic relationship with other Hepatozoon spp. AB - To characterize phylogenetically the species which causes canine hepatozoonosis at two rural areas of Rio de Janeiro State, Brazil, we used universal or Hepatozoon spp. primer sets for the 18S SSU rRNA coding region. DNA extracts were obtained from blood samples of thirteen dogs naturally infected, from four experimentally infected, and from five puppies infected by vertical transmission from a dam, that was experimentally infected. DNA of sporozoites of Hepatozoon americanum was used as positive control. The amplification of DNA extracts from blood of dogs infected with sporozoites of Hepatozoon spp. was observed in the presence of primers to 18S SSU rRNA gene of Hepatozoon spp., whereas DNA of H. americanum sporozoites was amplified in the presence of either universal or Hepatozoon spp.-specific primer sets; the amplified products were approximately 600bp in size. Cloned PCR products obtained from DNA extracts of blood from two dogs experimentally infected with Hepatozoon sp. were sequenced. The consensus sequence, derived from six sequence data sets, were blasted against sequences of 18S SSU rRNA of Hepatozoon spp. available at GenBank and aligned to homologous sequences to perform the phylogenetic analysis. This analysis clearly showed that our sequence clustered, independently of H. americanum sequences, within a group comprising other Hepatozoon canis sequences. Our results confirmed the hypothesis that the agent causing hepatozoonosis in the areas studied in Brazil is H. canis, supporting previous reports that were based on morphological and morphometric analyses. PMID- 17134838 TI - Differential behavior of the sub-sites of cytochrome 450 active site in binding of substrates, and products (implications for coupling/uncoupling). AB - The cytochrome P450 catalyzes hydroxylation of many substrates in the presence of O(2) and specific electron transport system. The ternary complex S-Fe(+)O(2) with substrate and O(2) bound to their respective sites on the reduced enzyme is an important intermediate in the formation of the hydroxylating species. Then the active site may be considered as having two sub-sites geared for entirely different types of functionally relevant interactions. The two sites are the substrate binding site, the specific protein residues (Site I), and the L(6) position of the iron (Site II) to which O(2) binds upon reduction. In the ferric enzyme, when substrate binds to Site I, the low spin six-coordinated P450 is converted to the readily reducible high spin five coordinated state. Certain amines and OH compounds, such as products of P450-catalyzed reactions, can bind to Site II resulting in six coordinated inhibited complexes. Then the substrate and product interactions with the two sub-sites can regulate the functional state of the enzyme during catalysis. Product interactions have received very little attention. CYP101 is the only P450 in which X-ray and spectroscopic data on all three structures, the substrate-free, camphor-bound and the 5-exo-OHcamphor-bound are available. The substrate-free CYP101 is low spin and six-coordinated with a water molecule ligated at the L(6) position of the iron. The substrate camphor binds to Site I, and releases the L(6) water despite its inability to bind to this site, indicating that Site I binding can inhibit Site II ligation. The product 5-exo-OHcamphor in addition to binding to Site I, binds to Site II through its -OH group forming Fe-O bond, resulting in the low spin six coordinated complex. New temperature-jump relaxation kinetic data indicating that Site II ligation inhibits Site I binding are presented. It appears that the Site I and Site II function as interacting sub-sites. The inhibitory allosteric interactions between the two sub-sites are also reflected in the data on binding of the substrate camphor (S) in the presence of the product 5-exo-OH camphor (P) to CYP101 (E). The data are in accordance with the two-site model involving the ternary complex ESP. The affinity of the substrate to the product-bound enzyme as well as the affinity of the product to the substrate-bound enzyme decreased with increase in product concentration, which is consistent with mixed inhibition indicative of inhibitory allosteric interactions between the two sub-sites. Implications of these observations for coupling/uncoupling mechanisms are discussed in the light of the published findings consistent with the two-site behavior of the P450 active site. In addition, kinetic data indicating that the transient high spin intermediate may have to be taken into account for understanding how some P450s have been able to express appreciable hydroxylation activities in the absence of substrate-induced low to high spin transition, observable by the traditional static spectroscopy, are presented. PMID- 17134839 TI - A novel technique to determine pressure in pressure garments for hypertrophic burn scars and comfort properties. AB - Current recommendations state that pressure garments should be worn for up to 2 years for hypertrophic burn scars. Thermo-physiological properties of pressure garments were assessed by the thermo-physiological tests and a comparison of the fabric with a sportwool which is a single-jersey knitted fabric was performed. In this novel technique, it was aimed to determine the exact pressure of pressure garments on the applied body part. For the theoretical part of this study, the Laplace equation was used with an optimum pressure of 20 mmHg and a relationship between change in length versus circumference was calculated. To determine the change in length a ruler was prepared for each predetermined circumference value using this relationship. Grid printed fabric samples were prepared with an interval of 2 cm in width direction to be used for the calculation of mean pressure on a Mannequin Leg. The resultant mean pressure calculated experimentally on Mannequin Leg as 24 mmHg was compared to the optimum pressure of 20 mmHg. It was observed that the two values were not statistically significantly different. PMID- 17134840 TI - Neoadjuvant endocrine therapy in breast cancer. AB - The use of endocrine therapy is well established as a primary treatment for locally advanced breast cancer. However, despite the current popularity of neoadjuvant chemotherapy for operable tumours, there is relatively little published evidence for pre-operative endocrine therapy in operable disease, particularly outside of the elderly population. The wider use of aromatase inhibitors (AIs) has encouraged studies that compare the efficacy of AIs with tamoxifen in the neoadjuvant setting, but there remains a lack of comparison of neoadjuvant with adjuvant endocrine therapies. This review discusses the current evidence regarding primary endocrine therapy, along with the factors involved in choosing appropriate patients for neoadjuvant therapy and the current opinions on length of treatment time and measurement of response prior to surgery. PMID- 17134841 TI - Polycystic ovary syndrome in men: Stein-Leventhal syndrome revisited. AB - Polycystic ovary syndrome (PCOS), also referred to as Stein-Leventhal syndrome, is one of the most common endocrinopathies. It is characterized by hyperandrogenism, hyperinsulinemia, central obesity, polycystic ovaries, and anovulation. However, some of these manifestations, including the polycystic ovaries, are neither specific for the disorder, nor found in all affected individuals. PCOS appears to be due to one or more primary defects in the upstream gonadotropin/androgen and/or insulin pathway, with the polycystic ovaries being one of many downstream manifestations. Yet, the pathophysiology of PCOS is not completely elucidated. Since the primary defect underlying PCOS may be an upstream endocrine and/or metabolic disturbance, rather than a defect in the ovaries themselves, we hypothesize that this aberration can also arise in men and that the absence of polycystic ovaries in men with other stigmata of the disorder should not eliminate the diagnosis. Our hypothesis is supported by the observation that a genetic susceptibility to PCOS exists, and that PCOS-type manifestations are not limited to women. Indeed, male relatives may suffer from insulin resistance, obesity, diabetes mellitus, and cardiovascular disease. Therefore, recognition of this syndrome in men is important, since pharmacologic treatments identified for women with PCOS may alleviate metabolic problems related to insulin resistance and its sequelae in men with a similar underlying defect. We suggest that first-degree relatives of patients with PCOS should be examined not only for phenotypic features characteristic of PCOS but also for biochemical evidence of hyperinsulinemia and hyperandrogenism. In addition to examining these individuals for obesity, the women should be evaluated for hirsutism and the men should be screened for early-onset male-pattern alopecia and excess hairiness. Serologic evaluation should included the ratio of fasting levels of glucose to insulin, a glucose tolerance test, the free testosterone level and the sex hormone-binding globulin level. Finally, both male and female first-degree relatives of patients with PCOS should be tested for the underlying molecular defect(s) of this condition, once it is identified. As new treatments for PCOS emerge, e.g. insulin-sensitizing drugs, it will be important to determine if these treatments have beneficial effects on the metabolic symptoms and complications in all afflicted patients, regardless of gender. PMID- 17134842 TI - Glycine conjugated and free bile acids, aided by acid suppression, can easily enter the esophageal epithelium where they can activate oncogenes and eventually carcinogenesis. PMID- 17134843 TI - Artificial neural networks for prediction of lymph node status in breast cancer patients. PMID- 17134844 TI - Should it be safer to use a redox couple, both with (R)-alpha-lipoic acid combined with (R)-dihydrolipoic acid for avoiding prooxidant action of alpha lipoic acid? PMID- 17134845 TI - Leflunomide as an antiatherogenic drug. PMID- 17134846 TI - Concomitant use of antioxidants may decrease the efficacy of cisplatin in cancer patients. PMID- 17134847 TI - Delusional parasitosis and the dopamine transporter. A new insight of etiology? AB - Delusional parasitosis (DP) is a psychotic condition in which a person has the unshakeable and mistaken belief (delusion) and/or aberrant perception (hallucination) of being infested with parasites. The disorder will be usually classified in a primary DP-group without a detectable cause (so-called pure forms), while secondary DP-groups are associated with general organic conditions, psychiatric illnesses and drugs (substance induced). Etiology and pathophysiology of DP remain however unknown. In the present paper we hypothesize for the first time a decreased striatal dopamine transporter (DAT)-functioning (corresponding with an increased extracellular dopamine-level) as etiologic condition for DP (primary and secondary groups). The DAT as key regulator of the dopamine-reuptake in the human brain is well known (regulation of the extracellular dopamine concentration). It is a presynaptic plasma membrane protein highly dense represented in the striatum. The hypothesis of a decreased DAT-functioning as etiologic condition by DP is revealed in case reports which show that DAT inhibitors, such as cocaine, pemoline, methylphenidate and other amphetamine derivatives can induce the clinical expression of DP. Several other associated causes of secondary DP-groups (medications, parkinson, chorea huntington, multiple system atrophy, diabetes, cerebrovascular diseases, alcoholism, traumatic brain injury, hyperuricemia, human immunodeficiency virus, iron deficiency, schizophrenia, depression) suggest that the clinical expression of DP may be related to a decreased striatal DAT-functioning (blocking, reduced ligand binding, reduced density, reduced activity). Our examined DP-cases (2-females) show means of magnetic resonance imaging a structurally damaged striatum. Furthermore, we presume that by the primary DP-group, the physiologically age related decline of the DAT-density is pathologically elevated. Based on this hypothesis we show in the present paper the relation between DP and decreased striatal DAT-functioning, trying to give a new insight into the pathophysiologically mechanism involved. The hypothesis provides supporting evidence that increased levels of extracellular dopamine in the striatum of DP patients is likely to be the result of decreased DAT-functioning and not increased rates of release. The hypothesis can be investigated simply by dopamine transporter imaging in patients with DP. PMID- 17134848 TI - Angiotensin II, corticosteroids, type II diabetes and the metabolic syndrome. AB - Syndrome X, the Metabolic Syndrome, and type II diabetes are closely related diseases that share risk factors and symptoms, notably insulin resistance. Several factors have been proposed either to mediate the disease(s) or to be their causes, and most converge on the endocrine/paracrine functions of the adipocyte. A common feature of such systems is their relative autonomy from systemic negative feedback regulation, for example by the HPA axis. We draw particular attention to two such mechanisms, both of which are associated with, and can cause, insulin resistance: the extra-adrenal production of corticosteroids, and the tissue renin angiotensin system of the adipocyte. These show another feature: the inter-regulation of glucocorticoid action and the RAS by positive feedback. Cortisol enhances the expression of 11 beta-HSD 1, and also of angiotensinogen and angiotensin type 1 receptors. In turn, angiotensin can stimulate further corticosteroid production, from the adrenal and perhaps from extra-adrenal sources. The instability inherent in such positive loops could account for the progressive nature of the disease(s), suggesting ways to break the circle. PMID- 17134849 TI - Safety, tolerability and efficacy of levodopa-carbidopa treatment for cocaine dependence: two double-blind, randomized, clinical trials. AB - RATIONALE: The role of dopamine in cocaine abuse has been long recognized. Cocaine use can profoundly alter dopaminergic functioning through depletion of this monoamine and changes in receptor functioning. Based on these facts, levodopa (L-dopa) pharmacotherapy may be helpful in reducing or abolishing cocaine use. OBJECTIVE: The current studies sought to evaluate the safety, tolerability and efficacy of L-dopa as a treatment for cocaine dependence. METHODS: In Study 1, 67 cocaine-dependent subjects were randomized in a 5-week, double-blind, placebo-controlled safety trial. Subjects received either placebo, or 400 mg L-dopa plus 100 mg of the peripheral decarboxylase inhibitor, carbidopa, in a sustained-release preparation (Sinemet CR). In Study 2, 122 cocaine-dependent subjects were enrolled in a 9-week, randomized, double-blind, placebo-controlled trial to compare placebo to 400/100 mg and 800/200 mg L dopa/carbidopa treatments. Placebo or L-dopa were administered twice daily in both studies. RESULTS: L-dopa was well tolerated with similar retention and medication adherence rates compared to placebo. Only two side effects occurred more often in L-dopa-treated patients: nausea and dizziness. L-dopa lowered diastolic blood pressure in a dose-dependent fashion. In these trials, L-dopa had no effect on cocaine use, cocaine craving, or mood. CONCLUSION: These two studies demonstrate the safety and tolerability of L-dopa pharmacotherapy in cocaine dependent patients. No evidence for greater efficacy of L-dopa compared to placebo was observed. The possibility of enhancing treatment effects by combining L-dopa with other behavioral or pharmacological interventions is discussed. PMID- 17134850 TI - In vivo analysis of the acidic ribosomal proteins BmP1 and BmP2 of the silkworm Bombyx mori in the yeast Saccharomyces cerevisiae. AB - In the silkworm Bombyx mori the ribosomal stalk P-protein family consists of two low MW acidic proteins, BmP1 and BmP2, and of one higher MW protein, BmP0, as shown by electrophoretical and immunoblotting western blot analysis of purified ribosomes. Treatment of ribosomes with alkaline phosphatase followed by electrofocusing shifted the isoelectric points to higher pH, implying phosphorylation of the proteins. The cDNAs encoding BmP1 and BmP2 proteins were constructed and expressed in the Saccharomyces cerevisiae mutant strains defective in either the endogenous P1 or P2 proteins. The recombinant silkworm proteins could complement the absence of the homologous yeast proteins and were incorporated to the ribosomes of the transformed strains, helping the binding of the remaining endogenous acidic proteins, present in the cytoplasm in different extent. Thus, BmP1 was able to replace YP1alpha, preferentially binding YP2beta to the ribosome, while BmP2 replaced both yeast P2 proteins and induced the binding of both YP1alpha and YP1beta. PMID- 17134851 TI - Selection against LINE-1 retrotransposons results principally from their ability to mediate ectopic recombination. AB - LINE-1 (L1) retrotransposons constitute the most successful family of autonomous retroelements in mammals and they represent at least 17% of the size of the human genome. L1 insertions have occasionally been recruited to perform a beneficial function but the vast majority of L1 inserts are either neutral or deleterious. The basis for the deleterious effect of L1 remains a matter of debate and three possible mechanisms have been suggested: the direct effect of L1 inserts on gene activity, genetic rearrangements caused by L1-mediated ectopic recombination, or the retrotransposition process per se. We performed a genome-wide analysis of the distribution of L1 retrotransposons relative to the local recombination rate and the age and length of the elements. The proportion of L1 elements that are longer than 1.2 Kb is higher in low-recombining regions of the genome than in regions with a high recombination rate, but the genomic distributions of full-length elements (i.e. elements capable of retrotransposition) and long truncated elements were indistinguishable. We also found that the intensity of selection against long elements is proportional to the replicative success of L1 families. This suggests that the deleterious effect of L1 elements results principally from their ability to mediate ectopic recombination. PMID- 17134852 TI - High intrachromosomal similarity of retrotransposon long terminal repeats: evidence for homogenization by gene conversion on plant sex chromosomes? AB - Retrotransposons are ubiquitous in the plant genomes and are responsible for their plasticity. Recently, we described a novel family of gypsy-like retrotransposons, named Retand, in the dioecious plant Silene latifolia possessing evolutionary young sex chromosomes of the mammalian type (XY). Here we have analyzed long terminal repeats (LTRs) of Retand that were amplified from laser microdissected X and Y sex chromosomes and autosomes of S. latifolia. A majority of X and Y-derived LTRs formed a few separate clades in phylogenetic analysis reflecting their high intrachromosomal similarity. Moreover, the LTRs localized on the Y chromosome were less divergent than the X chromosome-derived or autosomal LTRs. These data can be explained by a homogenization process, such as gene conversion, working more intensively on the Y chromosome. PMID- 17134853 TI - Deciphering the regulatory mechanisms of abiotic stress tolerance in plants by genomic approaches. AB - Environmental constraints that include abiotic stress factors such as salt, drought, cold and extreme temperatures severely limit crop productivity. Improvement of crop plants with traits that confer tolerance to these stresses was practiced using traditional and modern breeding methods. Molecular breeding and genetic engineering contributed substantially to our understanding of the complexity of stress response. Mechanisms that operate signal perception, transduction and downstream regulatory factors are now being examined and an understanding of cellular pathways involved in abiotic stress responses provide valuable information on such responses. This review presents genomic-assisted methods which have helped to reveal complex regulatory networks controlling abiotic stress tolerance mechanisms by high-throughput expression profiling and gene inactivation techniques. Further, an account of stress-inducible regulatory genes which have been transferred into crop plants to enhance stress tolerance is discussed as possible modes of integrating information gained from functional genomics into knowledge-based breeding programs. In addition, we envision an integrative genomic and breeding approach to reveal developmental programs that enhance yield stability and improve grain quality under unfavorable environmental conditions of abiotic stresses. PMID- 17134854 TI - Sequence correlation between neighboring Alu instances suggests post retrotransposition sequence exchange due to Alu gene conversion. AB - Alu elements constitute 10% of the human genome. Alu mobilization is important in the evolution of the human genome. While retrotransposition is the primary pathway of Alu mobilization, Alu gene conversion has been postulated as a secondary pathway for Alu mobilization in human genome. However, the mode and tempo of Alu gene conversions remain a mystery due to lack of sensitive statistical methods. In this paper, we present the first study on sequence correlation between Alu instances, measured by the number of shared mutations away from the Alu consensus, or co-mutations. Our analysis reveals a significantly elevated co-mutation rate between Alu instances that are located in close proximity along a chromosome. This effect is more pronounced outside Alu subfamily diagnostic positions. This effect peaks among immediately adjacent Alu instances, diminishes quickly in increasing distances between Alu instances, and vanishes beyond 5000 bp. Our results suggest that this effect reflects post retrotransposition sequence exchanges between Alu instances, mainly due to Alu gene conversions. PMID- 17134855 TI - High expression of Cyp6g1, a cytochrome P450 gene, does not necessarily confer DDT resistance in Drosophila melanogaster. AB - Cytochrome P450 monooxygenases, a family of detoxifying enzymes, are thought to confer resistance to various insecticides including DDT. Daborn et al. [Daborn, P., Yen, J.L., Bogwitz, M., Le Goff, G., Feil, et al. 2002. A single p450 allele associated with insecticide resistance in Drosophila. Science 297, 2253-2256.] suggested that the Accord transposable element causes overexpression of a Cyp6g1 allele, which has spread globally and is the basis of DDT resistance in Drosophila melanogaster populations. To determine whether the same phenomenon also operates in other Drosophila strains, we investigated 91-R, 91-C, ry(506), Wisconsin, Canton-SH and Hikone-RH strains. While the LC(50) values for the 91-R and Wisconsin strains are 8348 microg and 447 microg of DDT, respectively, values for the other four strains range between 0.74 to 20.9 microg. As expected, the susceptible ry(506) and 91-C strains have about 16-33-fold lower levels of CYP6G1 mRNA than the resistant 91-R and Wisconsin strains. Surprisingly, CYP6G1 mRNA and protein levels in the Canton-SH and Hikone-RH strains are as high as in the two resistant strains, yet they are as susceptible as the 91-C strain. The susceptible phenotype of the Canton-SH and Hikone-RH strains is not due to mutation in the Cyp6g1 gene; sequence analysis showed that Cyp6g1 alleles of resistant and susceptible strains are very similar and cannot be classified into resistant and susceptible alleles. As observed by others, we also found that only the 5'-upstream DNA of overexpressing alleles of Cyp6g1 has an insertional DNA, which is similar to Accord and Ninja elements. To examine the role of Cyp6g1 in DDT resistance, we substituted the Cyp6g1 allele of the 91-R strain with the allele from the susceptible 91-C strain via recombination and synthesized three recombinant lines. All three lines lacked Accord insertion and showed low expression of Cyp6g1 like the 91-C strain, yet they were as highly resistant as the 91-R strain. We conclude a strain may not have to have Accord insertion in the Cyp6g1 gene and the Cyp6g1 itself may not have to be overexpressed for DDT resistance to occur. PMID- 17134856 TI - A novel non-coding DNA family in Caenorhabditis elegans. AB - Many repetitive elements, for example, SINEs, LINEs, LTR-retrotransposons and other SSRs are dispersed throughout eukaryotic genomes. To understand the biological function of these repetitive elements is of great current research interest. In this study, we report on the identification of a novel non-coding DNA family, designated CE1 family, in the nematode C. elegans genome. Some CE1 elements constituted a large palindrome sequence. The CE1 elements were interspersed at 95 sites in the C. elegans genome. Most of the CE1 elements were associated with, or were within, protein-coding genes. The sequence of the CE1 elements indicated that some could form a hairpin structure. One of the CE1 family, CE1(bs258), is located in the first intron of a novel gene, C46H11.6 which encodes a PDZ/DHR/GLGF domain protein. In gfp and lacZ reporter gene assays the CE1(bs258) element appeared to behave as an enhancer element for the expression of C46H11.6 but no effect on the expression of the opposite direction gene, pat-10 which encodes the body-wall muscle troponin C. The CE1(bs258) RNA transcript was detected by RT-PCR even when CE1(bs258) was located in an intron. We conclude that CE1 elements are involved in the expression of adjacent genes and are therefore selectively retained in the C. elegans genome. We discussed a biological function of the CE1(bs258) having many transcription factor-binding sites. PMID- 17134857 TI - Body mass index and gynecological factors as determinants of bone mass in healthy Moroccan women. AB - Several studies have shown that low body mass index (BMI) is associated with low BMD and fractures. However, the results that have been published from studies on reproductive factors and BMD are extremely controversial, with some demonstrating a beneficial effect, while others show a detrimental impact of these factors on bone mass. OBJECTIVE: To study the influence of several gynecological factors (years since menopause (YSM), age at menarche and gynecological age or reproductive life) simultaneously with anthropometric factors as determinants of bone mineral density (BMD) in healthy women older than 40. METHODS: BMD was determined by dual energy X-ray absorptiometry (DXA) at the lumbar spine and femurs in women aged >40 randomly chosen from the population of Rabat with a cluster sampling method. RESULTS: Four hundred and twenty-two healthy women older than 40 years were included in the study. The mean age was 57.2 years (8.4) [40 79] and the mean number of parities was 4.42 (2.9) [0-14]. Osteoporosis according to the classification of WHO (T-score or =C(20)) polyunsaturated fatty acids (omega3 LC-PUFA). Atlantic salmon (Salmo salar L.) parr were fed a control fish oil diet (FO) or one of 3 experimental diets with 100% canola oil (CO) 100% SDA oil (SO), and a 1:1 mix of CO and SDA oil (MX) for 42 days. There were no differences in the growth or feed efficiency between the four diets. However, there were significant differences in the fatty acid (FA) profiles of the red and white muscle tissues. Significantly higher amounts of SDA, eicosapentaenoic acid (20:5omega3, EPA), docosahexaenoic acid (22:6omega3, DHA) and total omega3 FA occurred in both red and white muscle tissues of fish fed SO and FO compared with those fed CO. Feeding SO diet resulted in omega3 LC-PUFA amounts in the white and red muscle being comparable to the FO diet. This study shows that absolute concentration (mug/g) of EPA, DHA and total omega3 have been maintained over 6 weeks for Atlantic salmon fed 14% SDA oil. The balance between increased biosynthesis and retention of omega3 LC-PUFA to maintain the concentrations observed in the SO fed fish remains to be conclusively determined, and further studies are needed to ascertain this. PMID- 17134929 TI - Molecular properties and tissue distribution of 30K proteins as ommin-binding proteins from diapause eggs of the silkworm, Bombyx mori. AB - We previously reported the purification of an ommin-binding protein (OMBP) from an acid-methanol extract of diapause eggs of the silkworm and that OMBP reacted with the anti-30K proteins antiserum. In order to clarify the relationship between OMBP and the 30K proteins, we attempted to determine the sequence of the N-terminal amino acid of OMBP, which was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). We observed ten protein spots of various isoelectric points; the spots corresponded with 30 kDa. Based on the sequence of the N-terminal amino acid (20 residues), the spots belonged to two kinds of 30K proteins (6G1 and 19G1), which are known as the major plasma proteins in the larval hemolymph of the silkworm. The proteins are expected to attach to polysaccharide because they reacted with concanavalin A and elderberry bark lectin. Immunohistochemical observations clarified that the proteins were localized in yolk granules and serosa in the diapause egg. These results suggest that OMBP is composed of 30K proteins which were modified with polysaccharides. In addition, the expression of 30K proteins mRNA was observed at early embryonic stage in diapause eggs by RT-PCR analysis. The 30K proteins as OMBP may play an important role in the transport and accumulation of tryptophan metabolites and ommochrome during the formation of serosa. PMID- 17134930 TI - Antioxidant metabolism of Xenopus laevis embryos during the first days of development. AB - Reactive oxygen species (ROS) are formed and degraded in all aerobic organisms, but their role during embryonic development has not yet been well established. In this paper, we report the activities of various enzymes involved in antioxidant metabolism during the first 7 days of embryonic development of Xenopus laevis embryos. During the first two days of development, embryo antioxidant metabolism is based on catalase and superoxide dismutase activities. Later, the glutathione system is activated, and the activity of all the enzymes involved increases. The results presented in this study, together with previously reported data, support the hypothesis that antioxidant defences may include enzymes that are genetically regulated, while the other systems that appear to be environmentally modulated become relevant later in development, probably to protect embryos from environmental and toxic factors. PMID- 17134931 TI - Arthropod D2 receptors positively couple with cAMP through the Gi/o protein family. AB - The pyloric network is an important model system for understanding neuromodulation of rhythmic motor behaviors like breathing or walking. Dopamine (DA) differentially modulates neurons within the pyloric network. However, while the electrophysiological actions of DA have been well characterized, nothing is known about the signaling events that mediate its effects. We have begun a molecular characterization of DA receptors (DARs) in this invertebrate system. Here, we describe the cloning and characterization of the lobster D(2) receptor, D(2 alpha Pan). We found that when expressed in HEK cells, the D(2 alpha Pan) receptor is activated by DA, but not other monoamines endogenous to the lobster nervous system. This receptor positively couples with cAMP through multiple Gi/o proteins via two discrete pathways: 1) a G alpha mediated inhibition of adenylyl cyclase (AC), leading to a decrease in cAMP and 2) a G beta gamma-mediated activation of phospholipase C beta (PLC beta), leading to an increase in cAMP. Alternate splicing alters the potency and efficacy of the receptor, but does not affect monoamine specificity. Finally, we show that arthropod D(2) receptor coupling with cAMP varies with the cellular milieu. PMID- 17134932 TI - [Benign schwannoma of the breast: about two cases]. AB - Schwannoma is a rare nervous tumor developed on schwan cells. Only 22 cases of breast schwannoma have been published since 2005. It usually appears as a breast lump having clinical and radiological characteristics suggestive of kindness. Its diagnosis is histological. Its treatment is surgical. Through two observations and a review of the literature we would try to remind the characteristics of this tumor. PMID- 17134933 TI - [Laparoscopy-assisted vaginal pelvic exenteration]. AB - OBJECTIVE: The aim of the study was to evaluate the feasibility, morbidity and survival outcome of laparoscopy-assisted vaginal pelvic exenteration. PATIENTS AND METHODS: Since 2000, 7 en-bloc pelvic exenteration combining a vaginal or perineal approach and laparoscopic approach have been performed in our cancer center associated with complex laparoscopic reconstruction. All patients but one received previous irradiation. Two patients underwent a total pelvic exenteration; three patients an anterior and middle exenteration; two patients a middle and posterior exenteration. Urinary system was reconstructed with an ileal loop in one case, with a hand-assisted laparoscopic Miami pouch in four cases. Reconstruction of the vagina was performed with an omental cylinder in three cases, with a gluteal thigh flap in one case. A colorectal anastomosis was performed in three patients, one patient had an end colostomy. A mini-laparotomy conversion was necessary in one case because of a pelvic side involvement to perform an intraoperative irradiation. RESULTS: Mean time of the procedure was 6.5 hours with peroperative bleeding less than 500 cm3. Four patients presented minor complications. No revision of the Miami pouch was necessary. Mean length of hospital stay was 27 days. The four patients with a Miami pouch were able to self catheterize at the time of discharge. Mean follow-up was 14 months. Four patients died of the disease (three were metastatic). One patient presented a local recurrence. Two patients are free of disease. DISCUSSION AND CONCLUSION: Laparoscopic or laparoscopy-assisted vaginal pelvic exenteration followed by reconstruction is feasible with curative intent in selected patients. PMID- 17134934 TI - Comparison of the performance of the activPAL Professional physical activity logger to a discrete accelerometer-based activity monitor. AB - The aim of this study was to assess the accuracy of the 'activPAL Professional' physical activity logger by comparing its output to that of a proven discrete accelerometer-based activity monitor during extended measurements on healthy subjects while performing activities of daily living (ADL). Ten healthy adults, with unrestricted mobility, wore both the activPAL and the discrete dual accelerometer (Analog Devices ADXL202)-based activity monitor that recorded in synchronization with each other. The accelerometer derived data were then compared to that generated by the activPAL and a complete statistical and error analysis was performed using a Matlab program. This program determined trunk and thigh inclination angles to distinguish between sitting/lying, standing and stepping for the discrete accelerometer device and amount of time spent on each activity. Analysis was performed on a second-by-second basis and then categorized at 15s intervals in direct comparison with the activPAL generated data. Of the total time monitored (approximately 60 h) the detection accuracies for static and dynamic activities were approximately 98%. In a population of healthy adults, the data obtained from the activPAL Professional physical activity logger for both static and dynamic activities showed a close match to a proven discrete accelerometer data with an offset of approximately 2% between the two systems. PMID- 17134935 TI - Space and the parietal cortex. AB - Current views of the parietal cortex have difficulty accommodating the human inferior parietal lobe (IPL) within a simple dorsal versus ventral stream dichotomy. In humans, lesions of the right IPL often lead to syndromes such as hemispatial neglect that are seemingly in accord with the proposal that this region has a crucial role in spatial processing. However, recent imaging and lesion studies have revealed that inferior parietal regions have non-spatial functions, such as in sustaining attention, detecting salient events embedded in a sequence of events and controlling attention over time. Here, we review these findings and show that spatial processes and the visual guidance of action are only part of the repertoire of parietal functions. Although sub-regions in the human superior parietal lobe and intraparietal sulcus contribute to vision-for action and spatial functions, more inferior parietal regions have distinctly non spatial attributes that are neither conventionally 'dorsal' nor conventionally 'ventral' in nature. PMID- 17134936 TI - Growing up to one's standard. AB - Plant organs grow to characteristic sizes and shapes that are dictated by the plant's genotype and the identity of the organ. Significant progress has been made in identifying and characterizing regulatory factors that promote organ growth, which act either on cell proliferation or on cell expansion. Their activity is antagonized by repressors of growth that limit organ size. Although the way in which that genes determine the identity of an organ modify its growth patterns is still unclear, initial links between growth regulators and patterning activities are being uncovered. As for the differences in organ size and shape between plant species, studies of natural variation are beginning to shed light on the underlying molecular changes. PMID- 17134937 TI - Sulforaphane as a promising molecule for fighting cancer. AB - A number of natural compounds with inhibitory effects on tumorigenesis have been identified from our diet. Several studies have documented the cancer-preventive activity of a significant number of isothiocyanates (ITCs), the majority of which occur in plants, especially in Cruciferous vegetables. The most characterized ITC is sulforaphane (SFN). SFN has received a great deal of attention because of its ability to simultaneously modulate multiple cellular targets involved in cancer development, including: (i) DNA protection by modulating carcinogen-metabolizing enzymes and blocking the action of mutagens; (ii) inhibition of cell proliferation and induction of apoptosis, thereby retarding or eliminating clonal expansion of initiated, transformed, and/or neoplastic cells; (iii) inhibition of neoangiogenesis, progression of benign tumors to malignant tumors, and metastasis formation. SFN is therefore able to prevent, delay, or reverse preneoplastic lesions, as well as to act on cancer cells as a therapeutic agent. Taking into account this evidence and its favorable toxicological profile, SFN can be viewed as a conceptually promising agent in cancer prevention and/or therapy. PMID- 17134938 TI - Synthesis and photophysical processes of a novel 1,10-phenanthroline-containing p conjugated chromophores and Zn(II) chelated complex. AB - A novel 1,10-phenanthroline-containing p-conjugated chromophore, 3,8-di-(1 naphthyl)-1,10-phenanthroline, was synthesized by Suzuki reaction and its chelated complex was prepared. The UV-visible spectroscopy and photoluminescent (PL) properties were investigated. The results show that when the solution of the complex in THF located in low concentration range (<10(-6) mol/L), the fluorescence emission exhibit only one emission band and the excimer is formed with gradual increase in concentration of the complex. The emission spectra of the complex show obvious solvent effect. In addition, the intermolecular action between the complex and fullerene (C60) were investigated. The fluorescence intensity of the complex is decreased and the maximal emission obvious red shifted with the increasing of the concentration of C60, which suggests that the strong interactions between the complex and C60 happen in the excited state. PMID- 17134939 TI - Reactivation of an occult hepatitis B virus escape mutant in an anti-HBs positive, anti-HBc negative lymphoma patient. AB - BACKGROUND: Hepatitis B virus (HBV) often persists after resolution, but its replication is suppressed by antiviral T cells. Immunosuppressive treatment may lead to viral reactivation and severe hepatitis. Early antiviral therapy prevents reactivation but some occult HBV infections are not easily detectable. RESULTS: Here we describe a patient with a progressive non-Hodgkin lymphoma who had probably not been vaccinated against HBV and, before immunosuppression, showed antibodies (anti-HBs) against the viral surface antigen (HBsAg) as the only possible marker of occult HBV infection. Under immunosuppression he developed viremia (>10(8)copies/mL). The virus exhibited three S gene mutations (L109R, C137W, G145R) which led to false negative HBsAg results and diminished binding of vaccine-induced anti-HBs. CONCLUSIONS: Reliable screening and monitoring of severely immunosuppressed patients for HBV should include, in addition to anti HBc and HBsAg, anti-HBs and sensitive HBV DNA assays. Furthermore, active vaccination or hepatitis B immune globulin may not protect against such mutants. PMID- 17134940 TI - Neonatal frequency discrimination in 250-4000-Hz range: electrophysiological evidence. AB - OBJECTIVE: The precision of sound frequency discrimination in newborn infants in the 250-4000-Hz frequency range was determined using the neonatal electrophysiological mismatch response (MMR), the infant equivalent of adult mismatch negativity (MMN). METHODS: The electroencephalogram (EEG) was recorded in 11 full-term sleeping newborn infants mostly in active sleep (67% of the time). Pure tones were presented through loudspeakers in an oddball paradigm with a 800-ms stimulus onset asynchrony (SOA). Each stimulus block contained a standard (p=0.76) of 250, 1000, or 4000Hz in frequency (in separate blocks) and deviants with a frequency change of either 5% or 20% of the standard (p=0.12 of each). RESULTS: A positive ERP deflection was found at 200-300ms from stimulus onset in response to the 20% deviation from the 250, 1000, and 4000Hz standard frequencies. The amplitude of the response in the 200-300ms time window was significantly larger for the 20% than 5% deviation. CONCLUSIONS: We observed in newborn infants automatic frequency discrimination as reflected by a positive MMR. The newborns were able to discriminate frequency change of 20% in the 250 4000-Hz frequency range, whereas the discrimination of the 5% frequency change was not statistically confirmed. SIGNIFICANCE: The present data hence suggest that the neonatal frequency discrimination has lower resolution than that in adult and older children data. PMID- 17134941 TI - Nurses in cancer care--stress when encountering existential issues. AB - Previous research related to stress among health care staff has highlighted several complex issues; however, a deeper analysis of the existential components has been lacking. The purpose of this paper is to study the stress of registered nurses who work with terminally ill and dying cancer patients, according to an earlier model developed by Ekedahl [2001. How can you bear the Challenge of Working at the Edges of Life and Death? Coping Processes with Hospital Chaplains Encountering Existential Confrontation: a Study in Psychology of Religion., Uppsala University, Thesis Uppsala]. The present study is qualitative and hypothesis-generating. The material analyzed are based on a life story approach and interviews carried out with 15 Swedish nurses working in hospices, oncology wards, and outpatient services for patients with advanced cancer. The nurses' stress levels ranged from low to severe, including multifaceted stress with existential dimensions. Different types of stress appear to be related with the individual, group, institutional, and cultural level. PMID- 17134942 TI - Disease burden and epidemiology of soil-transmitted helminthiases and schistosomiasis in Asia: the Japanese perspective. AB - The disease burden due to soil-transmitted helminthiases (STH) and schistosomiasis is not well documented in Asia. Both STH and schistosomiasis are chronic diseases but case detection is not easy because of the absence of clinical symptoms. STH and schistosomiasis are, however, endemic in Asia and their burden is significant. At the preparatory meeting for the Hashimoto Initiative in Japan in 1997, STH and schistosomiasis were categorized as Group 2 diseases. Parasitic infections in this category were well understood at the time but sophisticated control strategies were lacking. Japan has promoted comprehensive collaborative projects to reduce the burden of STH and schistosomiasis throughout Asia, creating an international network to collect epidemiological information and to implement and improve disease control, thus extending the school-based control method that had proved so successful in Japan. PMID- 17134943 TI - Beyond deworming: the promotion of school-health-based interventions by Japan. AB - Deworming bestows a variety of health and socioeconomic benefits and has been embraced by developing countries. To extend the beneficial impact of deworming, the Asian Centre of International Parasite Control (ACIPAC) project has carried out activities to link deworming with health-promoting school programs in the Greater Mekong Subregion (Cambodia, Laos, Myanmar, Thailand and Vietnam). ACIPAC has also conducted an integrated school-health-based program, including deworming and malaria education, under the umbrella of the health-promoting schools initiative. Implementing "beyond-deworming" efforts is now a practical challenge in the subregion. PMID- 17134944 TI - Cough and ventilatory adjustments evoked by aerosolised capsaicin and distilled water (fog) in man. AB - Airway receptors mediate cough and ventilatory adjustments. Simultaneous assessment of cough sensory-motor components and changes in breathing pattern may provide insights into the receptor(s) prevailingly stimulated by inhaled irritants. Nineteen subjects inhaled capsaicin and fog up to threshold concentrations for cough. Cough intensity, respiratory sensations and changes in breathing pattern induced by the two irritants were compared. Capsaicin and fog cough threshold values did not correlate. Coughing induced by both agents was preceded by qualitatively similar sensations and by significant increases in minute ventilation and respiratory drive due to selective increases in tidal volume (P<0.01). Cough intensity was similar with both agents. Cough frequency and the intensity of the urge to cough were higher with capsaicin (P<0.01). The lack of correlation between fog and capsaicin cough threshold values suggests differences in the neural mechanisms activated. The selective increase in tidal volume suggests prevailing involvement of rapidly adapting receptors. The stronger sensations evoked by capsaicin may contribute to the higher cough frequency observed with this agent. PMID- 17134945 TI - PEEP-induced changes in epithelial permeability in inbred mouse strains. AB - Inbred mouse strains have demonstrated a range of susceptibilities to inhaled environmental irritants. C57Bl/6J mice are highly susceptible while C3H/HeJ mice are resistant to ozone exposures, as assessed by lavaged protein. However, lavaged protein reflects a loss of both the endothelial and epithelial barrier. To determine whether basal differences exist in the epithelial barrier, we measured soluble tracer ((99m)technetium-diethylenetriamine pentaacetic acid, (99m)Tc-DTPA) clearance from the lung in spontaneously breathing, anesthetized mice and mice ventilated with increased lung volume with applied positive end expiratory pressure (PEEP; 1, 6, or 10cmH(2)O). Both strains showed more rapid clearance during ventilation with 10cmH(2)O PEEP compared with other ventilation pressures (p<0.001). There was a substantial difference in clearance between the two strains during ventilation with 10cmH(2)O PEEP (mean half time for C57Bl/6J mice=19+/-4min versus 34+/-3min for C3H/HeJ mice; p<0.001). Thus, when lung volume is increased, the susceptible C57Bl/6J strain shows a greater change in epithelial barrier than the resistant C3H/HeJ strain. These results may reflect fundamental differences in lung architecture. PMID- 17134946 TI - Inspiratory muscles do not limit maximal incremental exercise performance in healthy subjects. AB - We investigated whether the inspiratory muscles affect maximal incremental exercise performance using a placebo-controlled, crossover design. Six cyclists each performed six incremental exercise tests. For three trials, subjects exercised with proportional assist ventilation (PAV). For the remaining three trials, subjects underwent sham respiratory muscle unloading (placebo). Inspiratory muscle pressure (P(mus)) was reduced with PAV (-35.9+/-2.3% versus placebo; P<0.05). Furthermore, V(O2) and perceptions of dyspnea and limb discomfort at submaximal exercise intensities were significantly reduced with PAV. Peak power output, however, was not different between placebo and PAV (324+/ 4W versus 326+/-4W; P>0.05). Diaphragm fatigue (bilateral phrenic nerve stimulation) did not occur in placebo. In conclusion, substantially unloading the inspiratory muscles did not affect maximal incremental exercise performance. Therefore, our data do not support a role for either inspiratory muscle work or fatigue per se in the limitation of maximal incremental exercise. PMID- 17134947 TI - Future of antibody purification. AB - Antibody purification seems to be safely ensconced in a platform, now well established by way of multiple commercialized antibody processes. However, natural evolution compels us to peer into the future. This is driven not only by a large, projected increase in the number of antibody therapies, but also by dramatic improvements in upstream productivity, and process economics. Although disruptive technologies have yet escaped downstream processes, evolution of the so-called platform is already evident in antibody processes in late-stage development. Here we perform a wide survey of technologies that are competing to be part of that platform, and provide our [inherently dangerous] assessment of those that have the most promise. PMID- 17134948 TI - Strategies for analysis of glycoprotein glycosylation. AB - Glycoproteins are known to exhibit multiple biological functions. In order to assign distinct functional properties to defined structural features, detailed information on the respective carbohydrate moieties is required. Chemical and biochemical analyses, however, are often impeded by the small amounts of sample available and the vast structural heterogeneity of these glycans, thus necessitating highly sensitive and efficient methods for detection, separation and structural investigation. The aim of this article is to briefly review suitable strategies for characterization of glycosylation at the levels of intact proteins, glycopeptides and free oligosaccharides. Furthermore, methods commonly used for isolation, fractionation and carbohydrate structure analysis of liberated glycoprotein glycans are discussed in the context of potential applications in glycoproteomics. PMID- 17134949 TI - Development and in vitro characterization of sol-gel derived CaO-P2O5-SiO2-ZnO bioglass. AB - A CaO-P(2)O(5)-SiO(2)-ZnO bioglass was formed by the sol-gel technique and characterized by Raman spectroscopy, X-ray diffraction, energy dispersive X-ray analysis (EDXA) and scanning electron microscopy (SEM). The surface reactivity of the resultant glass-ceramic specimens was analyzed by immersion studies in simulated body fluid (SBF). SEM-EDXS and inductively coupled plasma atomic emission spectrometry techniques were used to monitor changes in the glass surface and SBF composition. Osteoblast cell culture experiments were performed to assess the biocompatibility and the alkaline phosphatase activity. Cell counts of the osteoblasts cultured on the bioglass samples were studied and compared with the polystyrene plates. The cells cultured on the bioglass disks consistently showed a higher alkaline phosphatase activity and cell counts compared to cells cultured on either polystyrene plates or the base CaO-P(2)O(5) SiO(2) bioglass. This was due to cell proliferation and differentiation promoted by the zinc-substituted bioglass. PMID- 17134950 TI - Regional trabecular morphology assessed by micro-CT is correlated with failure of aged thoracic vertebrae under a posteroanterior load and may determine the site of fracture. AB - INTRODUCTION: Spinal mobilization is commonly used in the treatment of patients with back pain, including individuals with osteoporosis. Previous data indicated that traditional predictors of skeletal failure-lateral or anteroposterior bone mineral density (BMD) by dual energy X-ray absorptiometry (DXA) or geometry of the spinous process or vertebral body-do not predict failure load during posteroanterior spinal mobilization. Morphological differences and inhomogeneities in BMD may have important effects on vertebral strength but integral BMD values by DXA cannot reflect these potentially important differences. We investigated the determinants of spinal fracture using muCT. MATERIALS AND METHODS: We measured failure load and failure site in 11 T5-8 cadaveric specimens (mean age 78 years) when a posteroanterior load was applied at the spinous process of T6 using a servohydraulic material testing machine. Radiography and CT scan were used to verify failure site. We observed no damage to the adjacent T7 vertebrae following the T6 posteroanterior failure test. The T7 vertebrae were sectioned to produce regional samples of the spinous process, the lamina and a vertebral body core. Each sample was scanned with muCT to measure bone microarchitectural parameters. We segmented and analysed four trabecular regions (spinous process base and middle, central lamina and central vertebral body). We used one-way repeated measures ANOVA to compare regions and computed Pearson correlations to assess the relation between PA failure load of T6 and the morphological parameters of T7. RESULTS: The BV/TV at the base or middle of the T7 spinous process (fracture sites), Tb.N and Tb.Th at the base were significantly correlated with posteroanterior failure load of T6 (BV/TV base: r=0.74, p=0.01; BV/TV middle: r=0.73, p=0.01; Tb.N base: r=0.64, p=0.03; Tb.Th base: r=0.65, p=0.03). The Tb.Th of the lamina was significantly greater than Tb.Th of the spinous process base (p=0.002). CONCLUSIONS: Whereas previous data indicated that BMD by DXA was not a good predictor of posteroanterior failure load, regional BV/TV of the spinous process base and middle regions, the sites of fracture, are correlated with posteroanterior failure load. Trabecular thickness differed significantly between the base of the spinous process and the lamina, and may have influenced the site of fracture. PMID- 17134951 TI - Decrease in blood triglycerides associated with the consumption of eggs of hens fed with food supplemented with fish oil. AB - BACKGROUND: n-3 polyunsaturated fatty acids (n-3 PUFA) convey several health benefits, including a reduction of serum concentration of triglycerides (TG). AIM: To examine the effect on blood lipids, particularly TG, of a diet with n-3 PUFA enriched eggs in healthy volunteers in the Seychelles (Indian Ocean). METHODS: Double-blind crossover trial with one group of volunteers fed with 5 normal eggs per week during 3 weeks followed by 5 enriched eggs per week during the next 3 weeks while the other group received eggs in the inverse sequence. Hen feed was supplemented at 5% with tuna oil. Enriched eggs contained nine times more n-3 PUFA than usual eggs (mainly docosahexaenoic acid). RESULTS: Twenty-five healthy volunteers participated in the study. Based on pooled results observed during the two 3-week periods, consumption of enriched eggs was associated with a significant 16-18% decrease in serum triglycerides (P<0.01) but with no significant difference in serum LDL-cholesterol and HDL-cholesterol. Serum LDL cholesterol increased during the first 3-week period and decreased during the second 3-week period with both normal and enriched eggs. Participants did not report a systematic preference for either type of eggs. CONCLUSIONS: Reasonable consumption of n-3 PUFA enriched eggs was associated with a significant decrease in serum triglycerides. These eggs could be a palatably acceptable source of n-3 PUFA. PMID- 17134952 TI - Effect of a single high-fat meal on endothelial function in patients with the metabolic syndrome: role of tumor necrosis factor-alpha. AB - BACKGROUND AND AIMS: A single high-fat meal may induce endothelial activation and dysfunction in both normal subjects and in patients with type 2 diabetes. The aim of this study was to assess the effect of a high-fat meal on endothelial function in patients with the metabolic syndrome. METHODS AND RESULTS: Twenty-five patients with the metabolic syndrome (ATP III criteria) were matched for sex, age and body mass index with 25 subjects without the metabolic syndrome. All subjects ate under supervision a high fat meal (760 calories) with 59% energy from fat, 12% energy from protein and 29% energy from carbohydrates. Compared with the control group, subjects with the metabolic syndrome had reduced endothelial function, as assessed with the l-arginine test, and higher circulating levels of TNF-alpha. Following the high-fat meal, both triglyceride and TNF-alpha levels increased more in subjects with the metabolic syndrome than in subjects without, while endothelial function decreased more in subjects with the metabolic syndrome. There was a significant relation between increases in TNF-alpha levels and decreases in endothelial function score in subjects with the metabolic syndrome (r=-0.39, P=0.03). CONCLUSION: TNF-alpha levels are increased in subjects with the metabolic syndrome; moreover, a high-fat meal produces further increase in its levels associated with endothelial dysfunction. PMID- 17134953 TI - The dietary antioxidant resveratrol affects redox changes of PPARalpha activity. AB - BACKGROUND AND AIMS: Gene-environment interaction is behind the pathogenesis of most widespread diseases, and nutrition is among the environmental factors with the highest impact on human health. The mechanisms involved in the interaction between nutritional factors and the genetic background of individuals are still unclear. The aim of this study was to investigate whether resveratrol (RES), an antioxidant polyphenol of red wine, can influence the activity of PPARalpha in the rat hepatoma cell line McArdle-RH7777. PPARalpha is a transcriptional factor that regulates gene expression when activated by endogenous or exogenous long chain fatty acids. Its activation results in significant protection from cardiovascular diseases in humans. METHODS AND RESULTS: By means of the electromobility shift assay (EMSA), we observed that PPARalpha is redox-sensitive as it displays reduced DNA-binding activity following in vivo treatment of the cells with 1mmol/L diethylmaleate (DEM), a glutathione-depleting agent. This finding could be relevant considering the important role of redox balance in pathological and physiological processes. We also observed a dual effect of 100mumol/L RES on PPARalpha activity: it was able to prevent, to a large extent, the DEM-induced reduction of DNA-binding activity at earlier time points, when the effect of DEM was stronger, but it depressed PPARalpha activity at later time points, when the effect of DEM was greatly reduced. CONCLUSION: A nutritional substance, such as RES, is able to influence the activity of gene-regulating factors, but the net effect is difficult to predict when the compound involved has multiple biological properties. Caution is therefore warranted before drawing conclusions about the potential benefits of RES for human health. PMID- 17134954 TI - Low plasma levels of oxygenated carotenoids in patients with coronary artery disease. AB - BACKGROUND AND AIMS: Low circulating levels of carotenoids have been associated with cardiovascular disease. The distribution of different carotenoids in blood may have an impact on the cardioprotective capacity. The aim of the present study was to determine the plasma levels of 6 major carotenoids in patients with coronary artery disease (CAD) and relate the findings to clinical, metabolic and immune parameters. METHODS AND RESULTS: Plasma levels of oxygenated carotenoids (lutein, zeaxanthin, beta-cryptoxanthin) and hydrocarbon carotenoids (alpha carotene, beta-carotene, lycopene) were determined in 39 patients with acute coronary syndrome, 50 patients with stable CAD and 50 controls. Serological assays for inflammatory activity and flow cytometrical analysis of lymphocyte subsets were performed. Both patient groups had significantly lower plasma levels of oxygenated carotenoids, in particular lutein+zeaxanthin, compared to controls. Low levels of oxygenated carotenoids were associated with smoking, high body mass index (BMI), low high density lipoprotein (HDL) cholesterol and, to a minor degree, inflammatory activity. Plasma levels of lutein+zeaxanthin were independently associated with the proportions of natural killer (NK) cells, but not with other lymphocytes, in blood. CONCLUSION: Among carotenoids, lutein+zeaxanthin and beta-cryptoxanthin were significantly reduced in CAD patients independent of clinical setting. The levels were correlated to a number of established cardiovascular risk factors. In addition, the relationship between NK cells and lutein+zeaxanthin may indicate a particular role for certain carotenoids in the immunological scenario of CAD. PMID- 17134955 TI - Aminotransferase activity in morbid and uncomplicated obesity: predictive role of fasting insulin. AB - BACKGROUND AND AIMS: An elevation in liver enzymes and, most notably, high serum alanine aminotransferase (ALT) activity, has been correlated with metabolic syndrome and obesity. However, whether obesity per se or obesity-related co morbidities affect aminotransferase activity is still unclear. In this study we sought to evaluate serum aminotransferase activity in morbid and uncomplicated obese subjects METHODS: In this cross-sectional study, serum aminotransferase activity, anthropometric and metabolic parameters were assessed in 290 morbid and 105 uncomplicated consecutive obese subjects matched for body mass index (BMI) (40.1+/-6.8 vs. 39.9+/-8.3 kg/m(2), respectively), age (35.9+/-10 vs. 34.8+/-9.6 years, respectively), sex distribution and duration of obesity. RESULTS: Uncomplicated obese subjects showed significantly lower serum ALT activity (17.58+/-6.3 (range 10-39) vs. 23.43+/-16 (range 12-89) U/l, (p<0.001)), and lower aspartate aminotransferase (AST), AST/ALT ratios and gamma glutamyltranspeptidase (gammaGT) (p<0.01 for all) than morbid obese subjects. Only 11% women and 19% men in the uncomplicated obese group showed high ALT levels, while ALT activity was high in 48% women and 51% men in the morbid obese group. Fasting insulin was the best correlate of ALT activity (R(2)=0.21, p=0.003). CONCLUSIONS: Our findings show that elevated ALT and AST activity are associated with increased fasting insulin and not with obesity per se, suggesting that the presence of insulin resistance, rather than BMI alone, plays a role in mediating the increased aminotransferase activity. PMID- 17134956 TI - Effects of rosuvastatin on 3-nitrotyrosine and aortic stiffness in hypercholesterolemia. AB - BACKGROUND AND AIMS: Early atherosclerosis is characterized by reduced large artery distensibility, paralleled by an increased peroxynitrite formation and nitration of tyrosine in proteins. The aim of the present study was to investigate the short-term effect of cholesterol lowering with rosuvastatin on 3 nitrotyrosine (3-NT), a marker of peroxynitrite-mediated oxidative stress, and on arterial stiffness. METHODS AND RESULTS: 71 outpatients with primary hypercholesterolemia were recruited for this randomized open-label intervention study; 35 patients were assigned to 4-week rosuvastatin therapy (10mg daily) with a low-fat diet, and 36 patients to a low-fat diet only. Within the cohort of 71 hypercholesterolemic patients, there was a significant correlation between cholesterol levels, 3-NT and aortic pulse wave velocity (aPWV), that is a reliable measure of aortic stiffness. Among those patients who received rosuvastatin, significant reductions in plasma cholesterol, 3-NT and aPWV were observed. Reductions in both aPWV and 3-NT levels correlated significantly with the decrease in plasma cholesterol. Reduction of plasma cholesterol was the only independent predictor for reduced arterial stiffness following rosuvastatin therapy. CONCLUSION: Cholesterol reduction achieved following short-term rosuvastatin therapy is associated with a decrease in peroxynitrite-mediated oxidative stress and an improvement in large artery distensibility; reduction in arterial stiffness is directly attributable to rosuvastatin-induced cholesterol lowering and not to reduction of plasma 3-NT levels. PMID- 17134957 TI - Development of abdominal fat and incipient metabolic syndrome in young healthy men exposed to long-term stress. AB - BACKGROUND AND AIM: The sympathetic nervous system may be involved in the pathophysiology of insulin resistance and metabolic cardiovascular syndrome in young men. The aim was to study the effects of long-term stress on different features of the metabolic syndrome (MES) in formerly non-obese healthy young males during 5 months of defined conditions. METHODS AND RESULTS: Sixteen healthy male sailors (mean age 36.5 (SD)+/-7 years) participating in a sailing race around the world were recruited for the study. Investigations were done before the start and at stop overs after finishing laps 1, 2 and 4 (1, 2(1/2) and 5 months, respectively). Anthropometric and blood pressure data as well as biochemical data associated with MES were substantiated. Food intake and exercise were chartered and largely controlled. A mean weight loss of 4.5+/-2 kg (P<0.005), comprising both fat and lean body mass, was recorded during the first lap. Subsequently after 5 months, a weight gain, mainly consisting of 1.2+/-1.1 kg body fat (P<0.05), took place, concomitantly with a protein mass drop of 0.6+/ 1.1 kg (P<0.05). The body fat gain accumulated on the abdominal region. Elevated blood levels of HbA1c, insulin and the triglycerides/high-density lipoprotein ratio were also observed during the race. Likewise heart rate and systolic blood pressure increased slightly but to a statistically significant extent. CONCLUSIONS: Non-obese healthy young men exposed to long-term stress developed abdominal obesity and signs of a metabolic syndrome in embryo, also emphasized by biochemical and blood pressure alterations. It is suggested that long-term and sustained stress activation might be an additional risk factor for the development of MES, even after control of dietary and exercise habits. PMID- 17134958 TI - Evidence for oxidative stress at elevated plasma thiol levels in chronic exposure to carbon disulfide (CS2) and coronary heart disease. AB - OBJECTIVES: Oxidative stress in plasma may be promoted by plasma thiols such as homocysteine. However, other thiols such as glutathione may also exert antioxidant effects in vitro and in vivo. To further investigate whether plasma thiols act as prooxidants or antioxidants, we compared plasma oxidative status in patients with coronary heart disease (CHD) and in subjects occupationally exposed to carbon disulfide (CS(2)). METHODS: Fifty-five subjects chronically exposed to CS(2), 53 CHD patients, and 52 healthy controls were examined. To assess plasma oxidative status, concentrations of thiobarbituric reactive substances (TBARS) and total antioxidative capacity (TAC), as well as ferritin and ceruloplasmin were determined. Antioxidative reserve was assessed by the determination of vitamine E, uric acid, superoxide dismutase, catalase, and glutathion peroxidase. In addition, protein and non-protein plasma thiol levels were measured. RESULTS: Patients in both groups had increased levels of plasma thiols as compared to controls: CS(2)-exposed subjects presented with increased levels of thiols associated with plasma proteins, whereas CHD patients presented with elevated total homocysteine and cysteine levels. TBARS were significantly increased and TAC was significantly decreased both in CS(2)-exposed subjects and in CHD patients. In addition decreased activity of glutathione peroxidase, an antioxidative enzyme inhibited by thiol-containing compounds, was noted in both groups. CONCLUSION: These results demonstrate that regardless of their metabolic origin increased thiols are associated with increased oxidative stress in plasma. PMID- 17134959 TI - Physical activity and blood lipids in rural and urban Tanzanians. AB - BACKGROUND: Physical inactivity and raised blood lipids are two powerful risk factors for coronary heart disease (CHD). Incidence and mortality from CHD are expected to increase in developing countries. However, studies on the prevalence of cardiovascular risk factors in Africa are rare. In this study we examined the level of physical activity and serum lipids in rural and urban Tanzanians. METHODS: Rural and urban inhabitants, n=985, mean age 43.8 years [SD, +/-8.9] were investigated. Physical activity level (PAL) was assessed by an interview administered questionnaire and blood samples were collected and analysed for serum lipids. RESULTS: The rural population (n=501) reported a substantially higher PAL than the urban population (n=484). They also had significantly lower mean weight, body mass index (BMI), T-cholesterol, LDL-cholesterol, and HDL cholesterol, T-cholesterol/HDL-cholesterol ratio, triglycerides and Apolipoprotein A-1. CONCLUSION: This study demonstrates that the urban Tanzanians have a considerably lower physical activity level and a more unfavourable lipid pattern than rural Tanzanians. These findings underline the importance of undertaking preventive measures to counter the increasing incidence of CHD in urban African populations. PMID- 17134960 TI - Orlistat increases serum paraoxonase activity in obese patients. AB - BACKGROUND AND AIM: Previous studies have demonstrated that oxidative stress is increased in obese patients. The high-density lipoprotein (HDL) associated human paraoxonase 1 (PON1) can inhibit low-density lipoprotein oxidation and has an antiatherogenic effect. Our objective was to assess the effects of orlistat therapy combined with diet on body mass index (BMI), waist circumference, lipid parameters, blood pressure, serum glucose level and PON1 activity. METHODS AND RESULTS: A longitudinal, multicenter, randomized study with and without orlistat treatment was performed. One hundred thirty nine otherwise healthy, obese subjects were divided in to two groups: 78 persons received orlistat (120 mg three times a day) combined with diet while 61 persons were kept on diet only. Anthropometrical parameters, serum lipid levels and PON1 activity were measured at baseline and after 6 months of treatment. BMI and waist circumference were reduced more pronouncedly in the orlistat group than in the control group. Patients receiving orlistat also had significantly greater improvements in fasting blood glucose levels and blood pressure. The orlistat-treated group showed a greater reduction in total cholesterol and triglyceride levels. In addition, the serum PON1 activity in these patients was significantly increased compared to the diet-only group. CONCLUSIONS: The 6-month treatment with orlistat had a beneficial effect on the lipid profile and improved the antioxidant status by increasing serum PON1 activity. However, because of the limited therapeutic effectiveness, obese patients with hypercholesterolemia should receive additional lipid lowering medications. PMID- 17134961 TI - Nongenomic activation of the GC-A enzyme by resveratrol and estradiol downstream from membrane estrogen receptors in human coronary arterial cells. AB - BACKGROUND AND AIM: Resveratrol (RSVL), a polyphenolic phytoestrogen in grapes, confers multifaceted cardiovascular benefits. The cellular and molecular basis of RSVL actions has been largely undefined until now. METHODS AND RESULTS: In human coronary smooth muscle cells (HCSMCs), RSVL markedly (3.2-fold) enhanced cGMP formation (t(1/2): 6.3 min, EC(50): 1.8 microM) and stimulated kinase-G activity (4-fold). By contrast, RSVL had no effect on cAMP or PKA activity in these cells. The RSVL-enhanced cGMP/kinase-G activity was not abrogated by the nitric oxide synthase-inhibitor (L-NMMA, 10 microM), or the soluble guanylyl cyclase (sGC) inhibitor (ODQ, 10 microM). In membrane preparations from HCSMCs, RSVL activated GC in the particulate-, but not in the soluble-membrane fraction. Similar effects were due to the specific particulate-GC-A agonist atrial natriuretic peptide (ANP, 0.1-1 microM). The combined effects of RSVL and ANP were competitive. By contrast, the selective GC-B agonist (BNP) showed no response on cGMP, whereas that for GC-C (guanylin) produced only slight increases in cGMP levels. Estradiol (E2) mimicked the effects of RSVL on cGMP, but showed a 46% lower maximal response. Combining E2 with RSVL showed a competitive, rather than an additive, response. Further, cGMP formation by RSVL or E2 was significantly attenuated by the pure estrogen receptor blocker, ICI-182,780 (10 microM). CONCLUSION: These findings are the first to link RSVL with pGC/kinase-G activation downstream from membrane ERs in the vasculature, thus substantiating its coronary protective effects, even in endothelium-disrupted coronary arteries. PMID- 17134962 TI - Finding genes that underlie cancer using genetic tools. PMID- 17134963 TI - Environmental estrogens, imitators of a thousand faces. PMID- 17134964 TI - Molecular biology in head and neck cancer. AB - Major changes in the treatment of head and neck cancer are possible today because of the knowledge that we have on the molecular biology of these tumors. Different pathways are active in the development of this cancer and field cancerization is a major problem for the cure in early stage disease. Epidermal growth factor signal transduction pathway is now the principal target for this disease. New therapeutic strategies such as monoclonal antibodies and small molecules have appeared, however no more than 20% of the patients have objective responses with these therapies. Consequently, new alternatives of treatment in the basis of the understanding of molecular biology are necessary to increase the number of patients that can be cured in the future. PMID- 17134965 TI - Gene expression regulation and cancer. AB - Gene expression is mostly controlled at the level of the transcription initiation. The transcription control regions of protein-encoding genes include: the core promoter, where RNA polymerase II binds, the proximal and distal promoter, responsible for gene expression regulation, and the enhancers and silencers. Chromatin represents an additional level of regulation of gene expression. The switching between inactive and active chromatin is closely related to the activity of histone-modifying enzymes and chromatin-remodelling complexes. Transcriptional activation of a gene requires the binding of specific transcription factors to regulatory DNA elements, the opening of the chromatin, the binding of Mediator, and the assembly of the preinitiation complex with RNA polymerase and RNA synthesis initiation. Transcription factors ultimately transduce the proliferation signals elicited by growth factors. Moreover, many human oncogenes encode for transcription factors, and some of them are prevalent in particular neoplasias (e.g., MYC, MLL, PML-RARa). Also, some of the most prominent tumor suppressors (e.g. p53) are transcription factors. PMID- 17134966 TI - What can nanotechnology do to fight cancer? AB - The marriage of physics, chemistry and biology at the namometric scale, nanotechnology, is a powerful technology which is predicted to have a large impacto on life sciences and particularly cancer treatment. In the following we will show some examples of applications which has already reached clinical treatments as new ideas which may positively influence the understanding, diagnosis and therapy of cancer. PMID- 17134967 TI - Diagnosis and current treatment of neurological paraneoplastic syndromes. AB - Neurological paraneoplastic syndromes (NPS) affect only 0.01% of cancer patients, chiefly those affected by lung, breast, ovarian and stomach cancer. They frequently cause major disability and produce limitations in patients' daily activities; the character of the disease is irreversible. Clinical suspicion is fundamental for an early diagnosis and it must be backed up by the specification of certain antibodies both present in blood and in cerebrospinal fluid (CSF). Conventional treatments are very inefficient at the time of treating these disorders; at present, the administration of immunoglobulins, immunosuppressors, chemotherapy agents and corticoids are under study, but so far results are not promising. The aim of this review is to analyze the variety of NPS and describe the findings concerning autoimmunity and treatments used at present. PMID- 17134968 TI - Informed consent in radiation oncology: is consenting easier than informing? AB - The patient's right to be informed has been universally recognized and reflected in the legal system of many countries. This right to correct and complete information on behalf of the patient and his admission to proceed with the recommended diagnostic or therapeutic procedure is formalized in the document commonly known as informed consent. Although the legal and bioethical considerations regarding this document have been exhaustively discussed and consensuated, its content continues to create certain doubts and uncertainties. The formal content and the manner in which the consent is obtained are the most difficult aspects. In this article, we analyze what should be included in the written informed consent, with regard to the totality of the information which the patient receives, who should inform, and how the consent should be obtained, as well as how to reflect the different aspects of the variety of radiotherapeutic procedures in the informed consent. PMID- 17134969 TI - Bortezomib induces different apoptotic rates in B-CLL cells according to IgVH and BCL-6 mutations. AB - BACKGROUND: B-cell chronic lymphocytic leukemia (B-CLL) is a remarkably heterogeneous disorder. Some patients have an indolent disease whereas others undergo a more agressive presentation needing treatment. New therapeutics approaches are necessary for the treatment of B-CLL. Bortezomib (Btz), is a proteasome inhibitor, currently undergoing clinical trials whose function, at least in part, by stabilizing the IkappaBalpha protein and inhibiting NFkappaB activation. OBJECTIVE: The objective of this work was to study the effects of Btz on isolated human B-CLL cells, in vitro, and to correlate the differential rates of apoptosis induction with biological variables. MATERIAL AND METHODS: 31 B-CLL samples, from patients in stage A of Binet were used for this study, and the apoptotic effect of Btz on these cells was measured. RESULTS: Our data show that Btz treatment of B-CLL cells induces apoptosis in a time and dose-dependent manner. The apoptosis induction is mediated in part by inhibition of NFkappaB and is dependent on caspases activation. Interesting, in IgVH mutated cells, Btz have statistically significant differences in their in vitro activity on B-CLL cells according to their BCL-6 mutational status. CONCLUSIONS: Btz is a promising pharmacologic agent for the treatment of B-CLL, but its efficacy seems to be related to IgVH and BCL-6 mutational status, therefore, it could be interesting to further investigate the mechanisms involved in the different behavior of the cells in response to apoptosis induction by this drug. PMID- 17134970 TI - HER2 (erbB-2)-targeted effects of the omega-3 polyunsaturated fatty acid, alpha linolenic acid (ALA; 18:3n-3), in breast cancer cells: the "fat features" of the "Mediterranean diet" as an "anti-HER2 cocktail". AB - BACKGROUND: Data derived from epidemiological and experimental studies suggest that alphalinolenic acid (ALA; 18:3n-3), the main omega-3 polyunsaturated fatty acid (PUFA) present in the Western diet, may have protective effects in breast cancer risk and metastatic progression. A recent pilot clinical trial assessing the effects of ALA-rich dietary flaxseed on tumor biological markers in postmenopausal patients with primary breast cancer demonstrated significant reductions in tumor growth and in HER2 (erbB-2) oncogene expression. HYPOTHESIS: The molecular mechanism by which ALA inhibits breast cancer cell growth and metastasis formation may involve a direct regulation of HER2, a well characterized oncogene playing a key role in the etiology, progression and response to some chemo- and endocrine therapies in approximately 20% of breast carcinomas. METHODS: Using HER2-specific ELISA, flow cytometry, immunofluorescence microscopy, Western blotting, RT-PCR and HER2 promoter reporter analyses, we characterized the effects of exogenous supplementation with ALA on the expression of HER2 oncogene, a master key player in the onset and metastasis formation of breast cancer disease. Metabolic status (MTT) assays were performed to evaluate the nature of the cytotoxic interaction between ALA and the humanized anti-HER2 monoclonal antibody trastuzumab (Herceptin). To study these issues we used BT-474 and SKBr-3 breast cancer cells, which naturally exhibit amplification of the HER2 oncogene. RESULTS: ALA treatment dramatically suppressed the expression of HER2-coded p185Her-2/neu oncoprotein as determined by ELISA, flow cytometry, immunofluorescence microscopy and immunoblotting techniques. Interestingly, ALA-induced down-regulation of p185Her-2/neu correlated with a transcriptional response as no HER2 mRNA signal could be detected by RT-PCR upon treatment with optimal concentrations of ALA (up to 20 microM). Consistent with these findings, ALA exposure was found to dramatically repress the activity of a Luciferase reporter gene driven by the HER2 promoter. Moreover, the nature of the cytotoxic interaction between ALA and trastuzumab (Herceptin) revealed a significant synergism as assessed by MTT-based cell viability assays. CONCLUSIONS: i) These findings reveal that the omega-3 PUFA ALA suppresses overexpression of HER2 oncogene at the transcriptional level, which, in turn, interacts synergistically with anti-HER2 trastuzumab- based immunotherapy. ii) Our results molecularly support a recent randomized double blind placebo-controlled clinical trial suggesting that ALA may be a potential dietary alternative or adjunct to currently used drugs in the management of HER2 positive breast carcinomas. iii) Considering our previous findings demonstrating the <> of the omega-6 PUFA linolenic acid (LA; 18:2n 6) and the <> of the omega-3 PUFA docosahexaenoic acid (DHA; 22:6n-3) and of the omega-9 monounsaturated fatty acid oleic acid (OA; 18:1n-9), it is reasonable to suggest that a low omega-6/omega-3 PUFA ratio and elevated MUFA levels, the two prominent <> of the <>, should be extremely efficient at blocking HER2 expression in breast cancer cells. PMID- 17134971 TI - Impact of computerised chemotherapy prescriptions on the prevention of medication errors. AB - OBJECTIVES: The authors sought to evaluate the impact of computerised chemotherapy prescription on the reduction of medication errors. The purpose of this study was to assess the incidence of errors present in electronic versus manual prescription. MATERIAL AND METHODS: The data gathered from computerised chemotherapy prescription sheets were submitted to a prospective analysis as cases of the intervention groups. The control group was comprised of the handwritten chemotherapy prescription sheets. Chemotherapy prescriptions for consecutive oncology patients were analysed by 2 independent examiners, who investigated errors of omission, commission, interpretation of dates, abbreviations and illegible handwriting. The proportion of treatment prescriptions containing one or more errors and the median of errors were calculated in order in both groups. RESULTS: At least one error was detected in 100% of the manual prescriptions and in 13% of computerised prescriptions (p < 0.001). The median of errors per computerised prescription was 0 (range: 0- 1), whereas in manual prescriptions the median was 5 (range: 1-12) (p < 0.001). Errors of omission were predominant in manual prescriptions. Errors of commission were limited to 1 case of unjustified cytostatic agent infra-dosage in a computerised prescription. This error was present in 3 cases in handwritten prescriptions and, in addition, 1 case of premedication drug substitution was detected. Errors of interpretation of the date, use of abbreviations and illegible handwriting were frequent among manual prescriptions and were absent from computerised prescriptions. CONCLUSIONS: Electronic chemotherapy prescription is a powerful tool. In this study it has been shown to decrease chemotherapy-related medication errors and ensure that safe chemotherapy practices were followed. PMID- 17134972 TI - Circulating tumoral cells lack circadian-rhythm in hospitalized metastasic breast cancer patients. AB - BACKGROUND: The relationship between breast cancer and circadian rhythm variation has been extensively studied. Increased breast tumorigenesis has been reported in melatonin-suppressed experimental models and in observational studies. OBJECTIVES: Circulating Tumor Cells (CTC) circadian- rhythm may optimize the timing of therapies. This is a prospective experimental study to ascertain the day-time and night-time CTC levels in hospitalized metastasic breast cancer (MBC) patients. MATERIAL AND METHODS: CTC are isolated and enumerated from a 08:00 AM and 08:00 PM blood collections. 23 MBC and 23 healthy volunteers entered the study. 69 samples were collected (23 samples at 08:00 AM and 23 samples at 08:00 PM from MBC; 23 samples from healthy volunteers). Results from two patients were rejected due to sample processing errors. No CTC were isolated from healthy volunteers. RESULTS AND DISCUSSION: No-differences between daytime and night-time CTC were observed. Therefore, we could not ascertain CTC circadian-rhythm in hospitalized metastasic breast cancer patients. PMID- 17134973 TI - Infiltrating ductal carcinoma and synchronous malignant phyllodes tumour. Diagnostic and therapeutic approaches. AB - Simultaneous presentation of breast cancer and malignant phyllodes tumour is rare. A female patient presented with a nodule in her left breast (infiltrating ductal carcinoma). On magnetic nuclear resonance another suspicious lesion (malignant phyllodes) was found in the right breast. Bilateral mastectomy was performed. Thirty two months later the patient is still free of disease. The approach to dealing with synchronous breast tumours should be the same as that normally used. PMID- 17134974 TI - Small bowel occlusion due to intussusception of a single metastasis of lung cancer. AB - The occurrence of small bowel intussusception due to metastatic lung cancer is very infrequent. We present a case of young man recently diagnosed of non small cell lung cancer that had a unique small intestine metastasis. We discuss the diagnostic and therapeutic issues. PMID- 17134975 TI - Malignant uveitis masking a lymphocytic lymphoma. AB - A multitude of diseases can present as posterior bilateral uveitis. In most cases, the cause of pericardial effusion can be determined, but in some instances, the cause is not apparent even after making a systematic and complete diagnostic evaluation. We report here an unusual case of a patient who had a B cell lymphochytic lymphoma, which presented as bilateral posterior uveitis. The diagnosis by biopsy is described, as is the role of multiple test in the diagnosis of bilateral uveitis. PMID- 17134976 TI - Intra-abdominal follicular dendritic cell sarcoma. AB - Follicular dendritic cell (FDC) sarcoma is a very rare condition. We report here an intra-abdominal FDC sarcoma occurring as a mass, dependent on the celiac and left gastric lymph chains, that was completely excised. Eighteen months after surgery a recurrence at the liver pedicle was detected by a CT-scan and fully resected; in order to prevent another disease relapse postoperative radiotherapy was given. PMID- 17134977 TI - Cerebral and parotid metachronous metastases from an ovarian carcinoma. PMID- 17134978 TI - Lentiviral vector-mediated gene transfer to endotherial cells compared with adenoviral and retroviral vectors. AB - Human immunodeficiency virus (HIV, lentivirus) vector has attractive features for gene therapy, including the ability to transduce non-dividing cells and long-term transgene expression. We have already reported that lentivirus vector can transduce well-differentiated rat cardiac myocytes. Endothelial cells (EC) are an attractive target for gene therapy, both for the treatment of cardiovascular disease and for the systemic delivery of recombinant gene products directly into the circulation. There are several reports regarding application of adenovirus and retrovirus based vectors to EC. However, there have been few reports which show the effect to lentivirus-mediated gene transfer efficiency, compared with adenovirus and retrovirus. In this study, bovine aortic endothelial cells (BAECs) were infected, in vitro, with these virus vectors. Transduction efficiency (TE) of beta-Gal gene transfer in BAECs by adenovirus, lentivirus, or retrovirus at MOI10 (Multiplicity of infection) (determined on Hela cells) is 69+/-11, 33+/-8, or 22+/-6% respectively. In adenovirus and lentivirus, almost 100% of BAECs were transduced at MOI 50. However, in retrovirus, TE showed only 48+/-6% at MOI 50 and no increase at MOI 100. The percentage of beta-Gal positive cells was decreased rapidly at longer passage of cells after being transduced by adenovirus. However, lentivirus and retrovirus showed sustained higher percentage of positive cells. Furthermore, transduction by lentiviral vectors had no significant effect on viability of BAECs. Our results indicate that lentivirus showed high-level and long term gene expression in BAECs. Lentivirus can be an effective vector for the ex vivo, genetically modified EC implantation and in vivo gene therapy. PMID- 17134979 TI - New and easy strategy for cloning, expression, purification, and characterization of the 5S subunit of transcarboxylase from Propionibacterium f. shermanii. AB - Methylmalonyl CoA-oxalacetate transcarboxylase (EC 2. 1. 3. 1) from Propionibacterium f. shermanii is a biotin dependent enzyme which transfers CO2 from methylmalonyl-CoA (MMCoA) to pyruvate via a carboxylated biotin group to form oxalacetate. It is composed of three subunits, the central cylindrical hexameric 12S subunit, the outer six dimeric 5S subunit, and the twelve 1.3S linkers. We here report the cloning, sequencing, expression, and purification of the 5S subunit. The gene was identified by matching the amino acid sequence with that of deposited in the NCBI database. For cloned 5S subunit sequence shows regions of high homology with that of pyruvate carboxylase and oxaloacetate decarboxylase. The gene encoding the 5S subunit was cloned into the pTXB1 vector. The expressed 5S subunit was purified to apparent homogeneity by a single step process by using Intein mediated protein ligation (IPL) method. The cloned 5S gene encodes a protein of 505 amino acids and of M(r) 55,700. PMID- 17134980 TI - Study of ibuprofen glucopyranoside derivative synthesis by Candida antarctica lipase in organic solvent. AB - The direct esterification of ibuprofen and methyl alpha-D-glucopyranoside in organic solvent by Novozym 435 was investigated in terms of the main variables controlling the process, including initial water activity (a(w), 0.05-0.75), incubation time, (0-168 h) and substrate concentration. The results showed that the lower initial aw values resulted in higher enzymatic activity and bioconversion yield. The most appropriate initial aw and incubation time were 0.06 and 144 h, respectively. The results also showed that the optimal ratio of ibuprofen to methyl alpha-D-glucopyranoside was 2.0. By optimizing these parameters, the yield increased about 50%. In addition, the product was confirmed to be methyl 6-O-(2'-(4'-isobutylphenyl) propionyl) D-alpha-glucopyranoside. PMID- 17134981 TI - Diphenylether and macrotriolides occurring in a fungal isolate from the antarctic lichen Neuropogon. AB - The fungus, Tritirachium sp. HKI 0317, was isolated from the Antarctic lichen Neuropogon sp. Fermentation of this strain, extraction of the culture broth, and preparative separation of produced compounds furnished 4-carboxy-5,5'-dihydroxy 3,3'-dimethyldiphenylether (1), macrosphelide A (2), and macrosphelide J (3). The structures were elucidated on the basis of MS and NMR measurements, and the previously published data for this compounds. PMID- 17134982 TI - Characterization of ionically bound peroxidases from apple (Mallus pumilus) fruits. AB - Ionically bound peroxidases (POD) were salt extracted from the pulp of four Indian apple varieties, i.e., Golden delicious HP, Golden delicious JK, Red delicious, and Royal delicious. They were precipitated with chilled ethanol. Thermal treatments of partially purified enzymes were given from 40-70 degrees C for 30 minutes. Golden delicious HP peroxidase showed thermostability at 60 degrees C, while three other peroxidases were observed at 50 degrees C. Phenolic compounds (i.e., caffeic acid, ferulic acids, p-coumaric acid, protocatechuic acid) and metal ions (i.e., Cu2+ and Fe2+) activated all apple peroxidases. However, Mn2+ inhibited the peroxidases from Golden delicious HP, Golden delicious JK, and Red delicious, and a substantial increase was observed in Royal delicious peroxidase. Mg2+ inhibited the peroxidases from Golden delicious HP and Red delicious, but marginal activation was reported in peroxidases from Golden delicious JK and Royal delicious. Zn2+ established stimulation in Golden delicious HP and Golden delicious JK peroxidases, but inhibition was observed in peroxidases in Red delicious and Royal delicious.. Methionine, proline, tryptophan, and valine stimulated all four apple peroxidases, but cysteine showed inhibition in Golden delicious JK. PMID- 17134983 TI - FT-IR spectrophotometric analysis of coenzyme Q10 (CoQ10) and its pharmaceutical formulations. AB - A Fourier transform infrared (FT-IR) spectrometric method was developed for the rapid, direct measurement of coenzyme Q10 (CoQ10) in different pharmaceutical products. Conventional KBr spectra were compared for the best determination of active substance in drug preparations. Lambert-Beer's law and two chemometric approaches, partial least squares (PLS) and principal component regression (PCR+) methods, were used in data processing. PMID- 17134984 TI - Repeated use of immobilized Gluconobacter oxydans cells for conversion of glycerol to dihydroxyacetone. AB - Dihydroxyacetone (DHA) is of great interest in the fine chemical and pharmaceutical industry; therefore, the discovery of suitable biocatalysts for the efficient production of it is very necessary. In the experiment, Gluconobacter oxydans was immobilized in polyvinyl alcohol (PVA). Various parameters of the immobilized cells were investigated. The results have shown that the optimal conversion conditions by the immobilized cells were at 30 degrees C and pH 6.0. The immobilized cells remained very active over the period of 14 days for storage and only lost 10% of its original activity. Repeated use of immobilized cells for conversion of glycerol to DHA was carried out in a 1.5 L stirred tank reactor, the average conversion rate was about 86%. Despite the high shear stress, bead shape was not affected, even after five consecutive conversion cycles. The regenerated biocatalyst could recover 90% of its initial activity. PMID- 17134985 TI - A highly efficient solvent-free synthesis of benzoxanthenes catalyzed by methanesulfonic Acid. AB - A series of 14-alkyl and aryl-14H-dibenzo[a, j]xanthenes have been synthesized through a one-pot condensation of beta-naphthol with alkyl or aryl aldehydes, in the presence of methanesulfonic acid in a solvent-free medium, with high yields. PMID- 17134987 TI - Synaptic organization of the mouse cerebellar cortex in organotypic slice cultures. AB - The cellular and synaptic organization of new born mouse cerebellum maintained in organotypic slice cultures was investigated using immunohistochemical and patch clamp recording approaches. The histological organization of the cultures shared many features with that observed in situ. Purkinje cells were generally arranged in a monolayer surrounded by a molecular-like neuropil made of Purkinje cell dendritic arborizations. Purkinje cell axons ran between clusters of small round cells identified as granule cells by Kv3.1b potassium channel immunolabelling. The terminal varicosities of the Purkinje cells axons enwrapped presumptive neurons of the cerebellar nuclei whereas their recurrent collaterals were in contact with Purkinje cells and other neurons. Granule cell axons established contacts with Purkinje cell somata and dendrites. Parvalbumin and glutamine acid decarboxylase (GAD) immunohistochemistry revealed the presence of presumptive interneurons throughout the culture. The endings of granule cell axons were observed to be in contact with these interneurons. Similarly, interneurons endings were seen close to Purkinje cells and granule cells. Whole cell recordings from Purkinje cell somata showed AMPA receptor-mediated spontaneous excitatory post-synaptic currents (sEPSCs) and GABAA receptor-mediated spontaneous inhibitory post-synaptic currents (sIPSCs). Similar events were recorded from granule cell somata except that in this neuronal type EPSPs have both a NMDA component and an AMPA component. In addition, pharmacological experiments demonstrated a GABAergic control of granule cell activity and a glutamatergic control of GABAergic neurons by granule cells. This study shows that a functional neuronal network is established in such organotypic cultures even in the absence of the two normal excitatory afferents, the mossy fibers and the climbing fibers. PMID- 17134988 TI - Iron and iron-responsive proteins in the cardiomyopathy of Friedreich's ataxia. AB - Hypertrophic cardiomyopathy is a common complication of Friedreich's ataxia (FRDA). Histological sections reveal abnormal cardiomyocytes, muscle fiber necrosis, reactive inflammation, and increased endomysial connective tissue. Scattered muscle fibers display perinuclear collections of minute iron-positive granules that lie in rows between myofibrils. Frataxin deficiency in FRDA causes mitochondrial iron dysmetabolism. We studied total iron and the iron-related proteins ferritin, mitochondrial ferritin, divalent metal transporter 1 (DMT1), and ferroportin in FRDA hearts by biochemical and histological techniques. Total iron in the left ventricular wall of FRDA patients (30.7+/-19.3 mg/100 g dry weight) was not significantly higher than normal (31.3+/-24.1 mg/100 g dry weight). Similarly, cytosolic holoferritin levels in FRDA hearts (230+/-172 microg/g wet weight) were not significantly elevated above normal (148+/-86 microg/g wet weight). The iron-positive granules exhibited immunoreactivity for cytosolic ferritin, mitochondrial ferritin, and ferroportin. Electron microscopy showed enhanced electron density of mitochondrial deposits after treatment with bismuth subnitrate supporting ferritin accumulation. The inflammatory cells in the endomysium were reactive for CD68, cytosolic ferritin, and the DMT1 isoform(s) translated from messenger ribonucleic acids containing iron-responsive elements (DMT1+). Progressive cardiomyopathy in FRDA is the likely result of iron catalyzed mitochondrial damage followed by muscle fiber necrosis and a chronic reactive myocarditis. PMID- 17134989 TI - Cerebellar efferent neurons in teleost fish. AB - In tetrapods, cerebellar efferent systems are mainly mediated via the cerebellar nuclei. In teleosts, the cerebellum lacks cerebellar nuclei. Instead, the cerebellar efferent neurons, termed eurydendroid cells, are arrayed within and below the ganglionic layer. Tracer injections outside of the cerebellum, which retrogradely label eurydendroid cells demonstrate that most eurydendroid cells possess two or more primary dendrites which extend broadly into the molecular layer. Some eurydendroid cells mostly situated in caudal portions of the cerebellum have only one primary dendrite. The eurydendroid cells receive inputs from the Purkinje cells and parallel fibers, but apparently do not receive inputs from the climbing fibers. Eurydendroid cells of the corpus cerebelli and medial valvula project to many brain regions, from the diencephalon to the caudal medulla. A few eurydendroid cells in the valvula project directly to the telencephalon. About half of the eurydendroid cells are aspartate immunopositive. Anti-GABA and anti-zebrin II antibodies that are known as markers for the Purkinje cells in mammals also recognize the Purkinje cells in the teleost cerebellum, but do not recognize the eurydendroid cells. These results suggest that the eurydendroid cells receive GABAergic inputs from the Purkinje cells. This relationship between the eurydendroid and Purkinje cells is similar to that between the cerebellar nuclei and Purkinje cells in mammals. The eurydendroid cells of teleost have both dissimilar as well as similar features compared to neurons of the cerebellar nuclei in tetrapods. PMID- 17134990 TI - Molecular and synaptic organization of GABAA receptors in the cerebellum: Effects of targeted subunit gene deletions. AB - GABAA receptors form heteromeric GABA-gated chloride channels assembled from a large family of subunit genes. In cerebellum, distinct GABAA receptor subtypes, differing in subunit composition, are segregated between cell types and synaptic circuits. The cerebellum therefore represents a useful system to investigate the significance of GABAA receptor heterogeneity. For instance, studies of mice carrying targeted deletion of major GABAA receptor subunit genes revealed the role of alpha subunit variants for receptor assembly, synaptic targeting, and functional properties. In addition, these studies unraveled mandatory association between certain subunits and demonstrated distinct pharmacology of receptors mediating phasic and tonic inhibition. Although some of these mutants have a profound loss of GABAA receptors, they exhibit only minor impairment of motor function, suggesting activation of compensatory mechanisms to preserve inhibitory networks in the cerebellum. These adaptations include an altered balance between phasic and tonic inhibition, activation of voltage-independent K+ conductances, and upregulation of GABAA receptors in interneurons that are not affected directly by the mutation. Deletion of the alpha1 subunit gene leads to complete loss of GABAA receptors in Purkinje cells. A striking alteration occurs in these mice, whereby presynaptic GABAergic terminals are preserved in the molecular layer but make heterologous synapses with spines, characterized by a glutamatergic-like postsynaptic density. During development of alpha1(0/0) mice, GABAergic synapses are initially formed but are replaced upon spine maturation. These findings suggest that functional GABAA receptors are required for long-term maintenance of GABAergic synapses in Purkinje cells. PMID- 17134991 TI - On the cerebello-cerebral interactions. AB - The question of which type of information and how it is being processed by the puzzling cerebellar circuitry remains open. Numerous works have highlighted and delineated the roles of cerebellar pathways in various parameters of motor control, such as timing of motor commands. Recent anatomical and functional data on a possible genuine cerebellar contribution in the processing of 'cognitive', behavioral and emotional information have not yet generated a consensus. Despite an apparent homogeneous and uniform cytoarchitecture, the actors of the cerebellar orchestra do play different roles depending on the anatomical inputs/outputs of the cerebellar regions. The numerous interactions between the cerebellum and the cerebral cortex remain a major field of research. Fundamental questions related to the cerebro-cerebellar networks, such as the modulation of corticomotoneuronal discharges in various contexts, have not been fully addressed, or only indirectly, with recent methods. Complexity of circuitries and non optimal theoretical frameworks continue to hamper our understanding of cerebellar operations. PMID- 17134992 TI - Neurobehavioral alterations in an adolescent following posterior fossa tumor resection. AB - The posterior fossa syndrome (PFS) consists of a variety of symptoms, including cerebellar mutism, behavioral disturbances and personality changes. We report longitudinal clinical, neuroradiological and neurobehavioral findings in a 19 year-old left-handed patient, diagnosed with attentional deficit hyperactivity disorder (ADHD) at the age of 12, who underwent posterior fossa tumor resection. Although the patient did not develop cerebellar mutism after surgery, marked apathy and emotional indifference, urinary retention, eye-lid apraxia and visual hallucinosis became apparent after a brief interval of normal functioning. Based on these findings it is argued that the PFS might be considered a semiological heterogeneous condition with variable clinical expressions. Long-term follow-up investigations revealed subtle, but significant cognitive and affective deficits, resembling the cerebellar cognitive affective syndrome in adults. As demonstrated by functional neuroimaging studies with SPECT, symptoms were associated with perfusional deficits in the anatomoclinically suspected supratentorial regions, reflecting the distant impact of the cerebellum on cognitive and affective functions. PMID- 17134993 TI - Motor control mechanism by the cerebellum. PMID- 17134994 TI - Characteristics, causes and evolutionary consequences of male-biased mutation. AB - Mutation has traditionally been considered a random process, but this paradigm is challenged by recent evidence of divergence rate heterogeneity in different genomic regions. One facet of mutation rate variation is the propensity for genetic change to correlate with the number of germ cell divisions, reflecting the replication-dependent origin of many mutations. Haldane was the first to connect this association of replication and mutation to the difference in the number of cell divisions in oogenesis (low) and spermatogenesis (usually high), and the resulting sex difference in the rate of mutation. The concept of male biased mutation has been thoroughly analysed in recent years using an evolutionary approach, in which sequence divergence of autosomes and/or sex chromosomes are compared to allow inference about the relative contribution of mothers and fathers in the accumulation of mutations. For instance, assuming that a neutral sequence is analysed, that rate heterogeneity owing to other factors is cancelled out by the investigation of many loci and that the effect of ancestral polymorphism is properly taken into account, the male-to-female mutation rate ratio, alpham, can be solved from the observed difference in rate of X and Y chromosome divergence. The male mutation bias is positively correlated with the relative excess of cell divisions in the male compared to the female germ line, as evidenced by a generation time effect: in mammals, alpham is estimated at approximately 4-6 in primates, approximately 3 in carnivores and approximately 2 in small rodents. Another life-history correlate is sexual selection: when there is intense sperm competition among males, increased sperm production will be associated with a larger number of mitotic cell divisions in spermatogenesis and hence an increase in alpham. Male-biased mutation has implications for important aspects of evolutionary biology such as mate choice in relation to mutation load, sexual selection and the maintenance of genetic diversity despite strong directional selection, the tendency for a disproportionate large role of the X (Z) chromosome in post-zygotic isolation, and the evolution of sex. PMID- 17134995 TI - Host shift and speciation in a coral-feeding nudibranch. AB - While the role of host preference in ecological speciation has been investigated extensively in terrestrial systems, very little is known in marine environments. Host preference combined with mate choice on the preferred host can lead to population subdivision and adaptation leading to host shifts. We use a phylogenetic approach based on two mitochondrial genetic markers to disentangle the taxonomic status and to investigate the role of host specificity in the speciation of the nudibranch genus Phestilla (Gastropoda, Opisthobranchia) from Guam, Palau and Hawaii. Species of the genus Phestilla complete their life cycle almost entirely on their specific host coral (species of Porites, Goniopora and Tubastrea). They reproduce on their host coral and their planktonic larvae require a host-specific chemical cue to metamorphose and settle onto their host. The phylogenetic trees of the combined cytochrome oxidase I and ribosomal 16S gene sequences clarify the relationship among species of Phestilla identifying most of the nominal species as monophyletic clades. We found a possible case of host shift from Porites to Goniopora and Tubastrea in sympatric Phestilla spp. This represents one of the first documented cases of host shift as a mechanism underlying speciation in a marine invertebrate. Furthermore, we found highly divergent clades within Phestilla sp. 1 and Phestilla minor (8.1-11.1%), suggesting cryptic speciation. The presence of a strong phylogenetic signal for the coral host confirms that the tight link between species of Phestilla and their host coral probably played an important role in speciation within this genus. PMID- 17134996 TI - Human parental effort and environmental risk. AB - Parental investment decisions depend on multiple factors, including the extent that parental care benefits offspring. Humans should show reduced parental effort in environments where parenting cannot improve offspring survival. Data from the standard cross-cultural sample are used to test this prediction. The results show that maternal care was inversely associated with famine and warfare, and also showed a quadratic association with pathogen stress, increasing as pathogen stress increased to moderate levels, but decreasing at higher levels. Age at weaning showed a similar quadratic relation with pathogens. The curvilinear associations between parental effort and pathogen stress may reflect that the saturation point of parental care is a function of environmental hazards. Paternal involvement was also inversely related to pathogen stress. The association between pathogens and paternal involvement was partially mediated by polygyny. In sum, maternal and paternal care appears to have somewhat different relations with environmental hazards, presumably owing to sex-specific tradeoffs in reproductive effort. PMID- 17134997 TI - Spatial and temporal dependencies of cross-orientation suppression in human vision. AB - A well-known property of orientation-tuned neurons in the visual cortex is that they are suppressed by the superposition of an orthogonal mask. This phenomenon has been explained in terms of physiological constraints (synaptic depression), engineering solutions for components with poor dynamic range (contrast normalization) and fundamental coding strategies for natural images (redundancy reduction). A common but often tacit assumption is that the suppressive process is equally potent at different spatial and temporal scales of analysis. To determine whether it is so, we measured psychophysical cross-orientation masking (XOM) functions for flickering horizontal Gabor stimuli over wide ranges of spatio-temporal frequency and contrast. We found that orthogonal masks raised contrast detection thresholds substantially at low spatial frequencies and high temporal frequencies (high speeds), and that small and unexpected levels of facilitation were evident elsewhere. The data were well fit by a functional model of contrast gain control, where (i) the weight of suppression increased with the ratio of temporal to spatial frequency and (ii) the weight of facilitatory modulation was the same for all conditions, but outcompeted by suppression at higher contrasts. These results (i) provide new constraints for models of primary visual cortex, (ii) associate XOM and facilitation with the transient magno- and sustained parvostreams, respectively, and (iii) reconcile earlier conflicting psychophysical reports on XOM. PMID- 17134998 TI - Quantitative measure of sexual selection with respect to the operational sex ratio: a comparison of selection indices. AB - Despite numerous indices proposed to predict the evolution of mating systems, a unified measure of sexual selection has remained elusive. Three previous studies have compared indices of sexual selection under laboratory conditions. Here, we use a genetic study to compare the most widely used measures of sexual selection in natural populations. We explored the mating and reproductive successes of male and female bank voles, Clethrionomys glareolus, across manipulated operational sex ratios (OSRs) by genotyping all adult and pup bank voles on 13 islands using six microsatellite loci. We used Bateman's principles (Is and I and Bateman gradients) and selection coefficients (s' and beta') to evaluate, for the first time, the genetic mating system of bank voles and compared these measures with alternative indices of sexual selection (index of monopolization and Morisita's index) across the OSRs. We found that all the sexual selection indices show significant positive intercorrelations for both males and females, suggesting that Bateman's principles are an accurate and a valid measure of the mating system. The Bateman gradient, in particular, provides information over and above that of other sexual selection indices. Male bank voles show a greater potential for sexual selection than females, and Bateman gradients indicate a polygynandrous mating system. Selection coefficients reveal strong selection gradients on male bank vole plasma testosterone level rather than body size. PMID- 17134999 TI - Most-downloaded abstracts and full-text views in 2005: a top-12 survey. PMID- 17135001 TI - Magnetic resonance enteroclysis in the diagnosis of small-intestinal Crohn's disease: diagnostic accuracy and inter- and intra-observer agreement. AB - PURPOSE: To evaluate the diagnostic accuracy and inter- and intra-observer agreement of magnetic resonance enteroclysis (MRE) in patients with or without Crohn's disease of the small intestine. MATERIAL AND METHODS: 60 consecutive patients with or without Crohn's disease examined with MRE were included. Two observers independently reviewed the MRE examinations, searching for 12 pathological signs. The reference standard was ileoscopy or surgery of the terminal ileum performed in 41 patients. RESULTS: Crohn's disease of the small intestine was found in 24 (40%) patients. MRE findings of increased intestinal wall thickness, intestinal wall enhancement, intestinal wall ulcer, and inflammatory activity of the terminal ileum showed high sensitivity, specificity, and positive and negative predictive values. Intestinal stenosis had sensitivities ranging from 43% to 100%, depending on the cut-off value. Inter- and intra-observer agreement was good or excellent for most pathological signs. However, observer agreement of intestinal wall edema was only fair and moderate. CONCLUSION: MRE evaluated Crohn's disease with a high diagnostic accuracy in the terminal ileum. Most MRE variables were evaluated with good or excellent observer agreement, indicating that the method was highly reproducible. Our study supports the notion that MRE is an appropriate method for diagnosing Crohn's disease. PMID- 17135002 TI - Percutaneous transluminal cutting-balloon angioplasty for hemodialysis access stenoses resistant to conventional balloon angioplasty. AB - PURPOSE: To examine the value of cutting-balloon percutaneous transluminal angioplasty (PTA) for hemodialysis access with residual stenosis after conventional balloon PTA. MATERIAL AND METHODS: Angioplasty with conventional balloons was performed on 48 hemodialysis access stenoses in 28 patients. If the balloon waist still remained at the rated burst pressure, the balloon was reinflated up to three times. Fifteen of 48 stenoses had residual stenoses of more than 30% after conventional balloon PTA. In these 15 stenoses, additional cutting-balloon PTA was performed. RESULTS: The mean residual percent diameter stenoses before and after conventional balloon PTA were 77.6+/-3.4% and 48.6+/ 8.5%, respectively. Additional cutting-balloon PTA decreased the mean residual percent diameter to 27.9+/-10.0%, and the cutting balloon was completely inflated without complication. In 12 patients, the 6-month primary patency rate (+/-SE) was 90.0% (9.5), and the 1-year primary patency rate (+/-SE) was 25.0% (14.8). CONCLUSION: Additional cutting-balloon PTA was found useful for reducing residual stenosis. PMID- 17135003 TI - Preoperative portal vein embolization with a mixture of gelatin sponge and iodized oil: efficacy and safety. AB - PURPOSE: To evaluate whether portal vein embolization (PVE) using a mixture of gelatin sponge (GS) pieces and iodized oil is safe and effective in inducing hypertrophy of the future liver remnants (FLR). MATERIAL AND METHODS: PVE was performed in 14 patients (eight male and six female, mean age 65 years, range 35 81 years) diagnosed with malignant liver tumor before surgery, whose FLR volumes were judged too small to allow for safe resection. Liver volume change, biochemical data change, complications related to PVE, and postoperative complications were retrospectively evaluated. RESULTS: PVE was successful in all patients, and there were no procedural complications. Absolute FLR volume and FLR/total liver volume (TLV) ratio increased by 102 cm3 and 8% (mean values), respectively. Planned hepatectomies were cancelled in three patients due to extrahepatic metastasis or bile duct infection. Five of the 11 patients (45%) who underwent hepatectomies had major postoperative complications. However, complications due to hepatic failure were not seen. In 10 patients, except one whose outcome was fatal outcome, the mean hospitalization days with and without major complications were 73 and 33 days, respectively. CONCLUSION: PVE using a mixture of GS and iodized oil seems to be effective and safe in inducing hypertrophy of the FLR. PMID- 17135004 TI - Assessment of takotsubo (ampulla) cardiomyopathy using iodine-123 metaiodobenzylguanidine scintigraphy. AB - PURPOSE: To evaluate the role of cardiac sympathetic innervation in patients whose clinical features consisted of chest pain, transient ST-segment elevation, left ventricular apical akinesis, minimal elevation of cardiac enzymes, and onset of symptoms shortly after a severe stress condition. MATERIAL AND METHODS: Five female patients, mean age 67+/-14 years, underwent thoracic 123I-MIBG (planar and SPECT) and 67Ga citrate (planar) scans within 5 days after the onset of symptoms. The 123I-MIBG myocardial washout rate between early (30 min) and delayed (3 hours) planar images was calculated. All patients presented findings consistent with takotsubo-like syndrome. Echocardiograms showed the characteristic wall motion pattern of significant apical dysfunction. Acute-phase coronary angiographies revealed a non-obstructive pattern. A peculiar apical akinesis and basal normokinesis were observed on the ventriculograms. RESULTS: Impairment of cardiac neuronal uptake of 123I-MIBG based on a reduction of the heart-to mediastinum uptake ratio was observed in all patients, while the washout rate was raised in four patients. All patients presented an apical uptake defect in the 123I-MIBG SPECT and planar images and a normal 67Ga scintigraphy. CONCLUSION: Our data indicate that ampulla cardiomyopathy (AC) is associated with a cardiac sympathetic innervation deficit characterized by a reduced global 123I-MIBG uptake and an apical uptake defect. The lack of 67Ga uptake in the acute phase of this syndrome indicates that AC is probably not associated with an inflammatory process. PMID- 17135005 TI - Hepatic intra-arterial injection of 3-bromopyruvate in rabbit VX2 tumor. AB - PURPOSE: To evaluate the antitumoral effects of an intra-arterial injection of 3 bromopyruvate (3-BrPA) on liver VX2 tumor in rabbits. MATERIAL AND METHODS: Twenty rabbits with surgically implanted liver VX2 tumors were used. The rabbits were divided into three groups: a control, a saline, and a 3-BrPA group. Four rabbits were not treated at all, and they served as the control group. The saline group (n = 6) received only intra-arterial saline injection. The 3-BrPA group (n = 10) received an intra-arterial injection of 3-bromopyruvate through the hepatic artery. The delivered amounts of 3-bromopyruvate were as follows: 25 ml of 0.5 mM in six rabbits, 25 ml of 1.0 mM in two rabbits, and 25 ml of 2.0 mM in two rabbits. Four days after intra-arterial injection, the rabbits were sacrificed and histopathologic analysis of the explanted livers was performed with comparison of the tumor necrosis ratio (a percentage of the necrotic area versus the entire tumorous area) in each group. RESULTS: The mean tumor necrosis ratio was 12.5+/-4.2%, 44.8+/-24.7%, and 49.4+/-14.3% in the control, saline, and 3 BrPA groups, respectively. Between the control and the saline group, and between the control and the 3-BrPA group the mean tumor necrosis ratio appeared to be significantly different (P<0.05). However, there was no statistical difference in the mean tumor necrosis ratio between the saline and the 3-BrPA group (P = 0.416). CONCLUSION: A single session of intra-arterial injection of 3-BrPA showed no better results in terms of tumor necrosis than that of saline injection in a rabbit VX2 tumor model. PMID- 17135006 TI - O-(2-[18F]fluoroethyl)-L-tyrosine (18F-FET) uptake in mouse thymoma cells, and its biodistribution in mice and human volunteers. AB - PURPOSE: To evaluate O-(2-[18F]fluoroethyl)-l-tyrosine (18F-FET) uptake in mouse malignant thymoma (EL4), and its biodistribution in mice and humans. MATERIAL AND METHODS: First, 18F-FET uptake in EL4 cells was examined in an in vitro study. Second, the kinetics of 18F-FET uptake and its biodistribution were examined in mice after subcutaneous injection of EL4 cells and complete Freund's adjuvant. Finally, the kinetics of 18F-FET uptake and its biodistribution in healthy human volunteers were examined. RESULTS: In an in vitro study, 18F-FET was extensively incorporated in EL4 cells. In an animal study, 18F-FET accumulation in normal organs peaked within 30 min postinjection. The mean ratios of 18F-FET uptake in tumors and in inflammatory lesions to that in muscle tissue at 60 min postadministration were 2.18 (range 2.00-2.29) and 1.04 (range 0.95-1.14), respectively. In a human study, static images were taken 60 min after 18F-FET administration. Mean standardized uptake values (SUVs) of the liver (1.52, range 1.38-1.71) and kidneys (1.90, range 1.74-2.24) were nearly equal or slightly higher than that of muscle tissue (1.19, range 0.99-1.33). CONCLUSION: This study demonstrates that 18F-FET accumulation in thymoma is significantly higher than in normal organs. 18F-FET could be a useful tracer for tumor imaging. PMID- 17135007 TI - Quantification of global left ventricular function: comparison of multidetector computed tomography and magnetic resonance imaging. a meta-analysis and review of the current literature. AB - Cardiac morbidity and mortality are closely related to cardiac volumes and global left ventricular (LV) function, expressed as left ventricular ejection fraction. Accurate assessment of these parameters is required for the prediction of prognosis in individual patients as well as in entire cohorts. The current standard of reference for left ventricular function is analysis by short-axis magnetic resonance imaging. In recent years, major extensive technological improvements have been achieved in computed tomography. The most marked development has been the introduction of the multidetector CT (MDCT), which has significantly improved temporal and spatial resolutions. In order to assess the current status of MDCT for analysis of LV function, the current available literature on this subject was reviewed. The data presented in this review indicate that the global left ventricular functional parameters measured by contemporary multi-detector row systems combined with adequate reconstruction algorithms and post-processing tools show a narrow diagnostic window and are interchangeable with those obtained by MRI. PMID- 17135008 TI - Magnetic resonance imaging and radiographic findings of seal finger. AB - PURPOSE: To describe the magnetic resonance imaging (MRI) and radiographic findings of five patients with seal finger. MATERIAL AND METHODS: The MR images and radiographs of five patients with seal finger were retrospectively evaluated. MRI was performed on four patients in the subacute phase, and follow-up imaging was done on one of them at 5 months. One patient had MRI only at a later stage 5 years after onset. Radiographs were taken three times in the subacute phase and once at a later stage. One patient had had seal finger in another finger previously. RESULTS: Short-tau inversion-recovery (STIR) sequence showed extensive subcutaneous soft tissue edema in all four patients in the subacute phase and tenosynovitis of the flexion tendons in two cases. Three patients had edema in 2-3 phalanges, and effusion in the distal interphalangeal (DIP) joint was seen in one case. At the later stage, no signal pathology in soft tissues or bones was seen in STIR images. In the subacute phase, radiographs showed digital soft-tissue swelling in three patients, and one patient had a narrowed DIP joint, periarticular osteoporosis, and a periosteal reaction. At the later stage, flexion contracture of the finger was seen. CONCLUSION: In addition to soft tissue infection, seal finger causes bone marrow edema, tenosynovitis, and effusion in the interphalangeal joints visible as increased signal intensity in STIR images. Radiographs reveal periarticular osteoporosis with loss of cartilage in the subacute phase and flexion contracture at the later stage. MRI (STIR) allows more precise delineation of the inflammatory process compared to radiography. PMID- 17135009 TI - Arrangement of fiber tracts forming Probst bundle in complete callosal agenesis: report of two cases with an evaluation by diffusion tensor tractography. AB - We report two patients with complete callosal agenesis in whom Probst bundles in both hemispheres could be depicted by diffusion tensor tractography (DTT). While one patient had no associated telencephalic anomaly other than callosal agenesis, the other had cortical dysplasia in the right frontal lobe. Although Probst bundles in the three normal hemispheres were well developed, that in the hemisphere which was affected by cortical dysplasia was small and poorly developed. DTT also showed that the fibers from the frontal pole ran more on the inner side of the Probst bundle than those from a more caudal region of the frontal lobe. Furthermore, fibers from the orbital gyri ran along the outermost side of Probst bundle. The arrangement of these fiber tracts in Probst bundle may reflect the developmental process of callosal fibers in their normal formation. PMID- 17135010 TI - Increased sensitivity to pathological brain changes using co-registration of magnetic resonance imaging scans. AB - PURPOSE: To compare automatic software-based co-registration of serial magnetic resonance imaging (MRI) scans with conventional visual comparison, by expert neuroradiologists. MATERIAL AND METHODS: Sixty-four patients who were referred to our epilepsy MRI unit for cerebral imaging were identified as having potentially, non-, or slow-growing lesions or cerebral atrophy and followed with sequential scans over a period of up to 8 years, resulting in a total of 92 pairs of scans. Scans were categorized as showing either lesions or atrophy. Each pair of scans was reviewed twice for the presence of change, with and without co-registration, performed using automated software. RESULTS: Co-registration and visual reporting without co-registration were discordant in the lesions group in nine out of 69 datasets (13%), and in 16 out of 23 pairs of scans in the atrophy group (69%). The most common cause of discordance was visual reporting not detecting changes apparent by co-registration. In three cases, changes detected visually were not detected following co-registration. CONCLUSION: In the group of patients studied, co-registration was more sensitive for detecting changes than visual comparison, particularly with respect to atrophic changes of the brain. With the increasing availability of sophisticated independent consoles attached to MRI scanners that may be used for image co-registration, we propose that serial T1-weighted volumetric MRI brain co-registration should be considered for integration into routine clinical practice to assess patients with suspected progressive disease. PMID- 17135011 TI - Balloon occlusion test of the internal carotid artery: correlation with stump pressure and 99mTc-HMPAO SPECT. AB - PURPOSE: To evaluate the correlation of stump pressure during balloon occlusion test and relative cerebral blood flow (relative CBF) as measured by 99mTc hexamethylpropyleneamine oxime (99mTc-HMPAO) single-photon emission computed tomography (SPECT) after test occlusion. MATERIAL AND METHODS: Balloon occlusion test of the internal carotid artery (ICA) was performed in 25 patients. The count ratio of occluded hemisphere to non-occluded hemisphere was calculated on 99mTc HMPAO SPECT. The ratio of mean stump pressure to mean arterial pressure during carotid occlusion during the balloon occlusion test was compared with the count ratio of 99mTc-HMPAO SPECT. RESULTS: Two patients failed to tolerate even brief carotid occlusion. The other 23 patients showed no ischemic deficit during occlusion of the ICA. In 13 of these 23 patients, the ratios of mean stump pressure to mean arterial pressure were more than 50%, and the count ratios on SPECT were more than 85%. In 10 of 23 patients, the ratios of mean stump pressure to mean arterial pressure were less than 50%, and the count ratios on SPECT were variable. CONCLUSION: Maintenance of a mean stump pressure of 50% or more of the mean systemic pressure during test occlusion indicates adequate cerebral blood flow during carotid occlusion. PMID- 17135012 TI - Sequential dynamic gadolinium magnetic resonance perfusion-weighted imaging: effects on transit time and cerebral blood volume measurements. AB - PURPOSE: To evaluate if two gadolinium perfusion studies can be performed during the same table occupancy without degradation of the derived data in the second study. MATERIAL AND METHODS: Magnetic resonance (MR) perfusion studies of the whole brain were performed on 12 patients during the administration of two gadolinium boluses separated by 8 min. In six patients, gadolinium was given as two 20-ml administrations of standard 0.5 M chelate (Magnevist), whilst the other six patients received two 10-ml administrations of 1.0 M chelate (Gadovist). RESULTS: There were no significant differences in subjective quality between the time-intensity curves of the first and second perfusion studies using either the 0.5 M or 1.0 M gadolinium chelate. The objective measurements in quality of the time-intensity curves (maximum signal change and full width at half maximum) changed by less than 5% of the original values on the second perfusion study. The first-moment mean transit times did not change significantly on the sequential studies. The regional cerebral blood volume tended to increase on the second study (by 15% on average), but this did not reach statistical significance. CONCLUSION: The results from two sequential dynamic gadolinium-based perfusion studies can be compared in a meaningful manner using the technique described. PMID- 17135013 TI - Corpus callosum hematoma secondary to isolated inferior sagittal sinus thrombosis. AB - A 45-year-old female was admitted with headache and vomiting. Cranial computed tomography (CT) demonstrated a callosal hematoma. Magnetic resonance imaging (MRI) showed no venous flow and thrombus replacing the inferior sagittal sinus (ISS) lumen. Under appropriate medical treatment and close follow-up she recovered quickly and, after 2 years, was doing well with corpus callosum infarcts. Isolated inferior sagittal sinus thrombosis is an extremely rare condition with only one previously reported case in the literature. Although it is very rare, isolated inferior sagittal sinus thrombosis should be considered in the differential diagnosis of non-traumatic corpus callosum hematoma. PMID- 17135014 TI - Novel diffusion-weighted magnetic resonance imaging findings in leptomeningeal carcinomatosis: a case report. AB - This report presents a rare case of leptomeningeal carcinomatosis initially presenting with mental impairment and rapidly progressing to coma without any history of malignancy. In addition to highlighting the diagnostic difficulties, the linear high signal intensity along the cortex on the diffusion-weighted imaging (DWI) sequence of magnetic resonance (MR) imaging was identified accidentally. High signal change in the corresponding areas was also noted on unenhanced fluid-attenuated inversion recovery (FLAIR) MR imaging, which may be a novel method of diagnosing leptomeningeal carcinomatosis, which should be studied further. PMID- 17135015 TI - Atypical imaging findings of infantile hemangioendothelioma: a case report. AB - Infantile hemangioendothelioma is the third most common hepatic tumor in children, and the most common benign vascular tumor of the liver in infancy. On computed tomography (CT) and magnetic resonance imaging (MRI), typical enhancement patterns similar to those seen in adult hemangiomas may help establish the diagnosis. However, atypical enhancement patterns may also occur. In this paper, we report a rare form of solitary infantile hemangioendothelioma that showed a rim-like calcification and rim-like enhancement on CT and MRI. PMID- 17135016 TI - Misrepresentation of surface rendering of pediatric brain malformations performed following spatial normalization. AB - PURPOSE: To evaluate the effects of spatial normalization on volume rendering in cases of pediatric brain malformation. MATERIAL AND METHODS: Three-dimensional (3D) T1-weighted volume datasets were acquired in three children, one with pachygyria, one with a Dandy-Walker malformation associated with polymicrogyria, and one with dysgenesis of the corpus callosum. On the non-normalized datasets, the skull margins were cropped and the remainder stripped with the brain extraction technique (BET). The data were also normalized into standard anatomic reference space using pediatric templates prior to the BET script. The surface constructions obtained by both techniques were then compared for geometric distortions. RESULTS: Normalization of 3D datasets resulted in significant distortions in the shape of the brain, with increased anterior-posterior dimensions and narrower transverse diameter in all three cases. In two cases, there were alterations in the appearance of the gyri and sulci, leading to a potential misinterpretation of the volume-rendered surface when the gyri and sulci were in fact normal. CONCLUSION: In pediatric brain, particularly those with congenital brain anomalies, normalization as a post-processing step should be avoided as this may lead to misrepresentation of brain morphometry. PMID- 17135017 TI - Diffusion-weighted magnetic resonance imaging of treatment-associated changes in recurrent and residual medulloblastoma: preliminary observations in three children. AB - PURPOSE: To emphasize a possible role of magnetic resonance (MR) diffusion weighted imaging (DWI) for lesion conspicuity and detection of treatment effects in children with medulloblastoma. MATERIAL AND METHODS: Three children with medulloblastoma (two residual and one recurrent) were examined repetitively by MR diffusion-weighted imaging. Regional assessment of the apparent diffusion coefficient (ADC) was done for tumorous lesions and periventricular white matter appearing normal on standard MR images. RESULTS: All lesions were clearly visible on DWI. In the case of recurrent tumor, on one scan, DWI showed lesions that were not seen on contrast-enhanced MRI. Increase (41%) of ADC was seen in one lesion, which subsequently responded completely to treatment over 27 months' follow-up. Intermediate increases (23-26%) of ADC were found with partial therapy response in three lesions. In contrast, a decrease (-11%) of ADC in two lesions was seen with tumor progression. CONCLUSION: These observations may suggest a role for DWI in early detection of metastatic disease and treatment monitoring of medulloblastoma, warranting a formal study. PMID- 17135018 TI - Role of magnetic resonance imaging in patient selection for uterine artery embolization. AB - Uterine fibroids are common tumors of the female pelvis. Uterine artery embolization (UAE) is an effective treatment of symptomatic uterine leiomyoma in the appropriate candidates, reducing or eliminating leiomyoma-related symptoms of bleeding, bulk, and/or pain. Magnetic resonance imaging (MRI) can be used to assess women with symptoms potentially attributable to uterine leiomyomas, and help to determine who is an appropriate candidate for UAE. Because of soft tissue characterization, multiplanar imaging capabilities, and enhancement, MR imaging not only accurately detects and characterizes uterine leiomyomas but also may predict who will benefit from the embolization. MRI ability to detect coexistent uterine or pelvic pathology may change the diagnosis and treatment management of patients being evaluated for UAE. PMID- 17135019 TI - Reduction in estimated vitamin A intake induced by new food composition tables in Japan, where vitamin A is taken mostly from plant foods. AB - A revised edition of the standard tables of food composition was implemented in Japan in 2005; one of the major revision points is the change of retinol activity equivalents for pro-vitamin A carotenoids. This preliminary analysis was conducted to examine whether the revision affects the estimation of vitamin A intake; and if so, to what extent. Accordingly, a field survey was conducted to collect 24-h duplicates of daily foods of citizens, and 26 adult women volunteered. Application of the procedures in the new and previous standard tables of food composition gave 537 microg retinol activity equivalent and 704 microg retinol equivalence, respectively, for daily vitamin A intake. Thus, the changes in retinol activity equivalents induced substantial reduction (by 24%) in estimation of vitamin A intake among the Japanese population, for whom pro vitamin A carotenoids in plant foods are the major sources (76%) for vitamin A, and retinol accounts for only 35% (on the retinol activity equivalent basis). PMID- 17135020 TI - The cupric ion reducing antioxidant capacity and polyphenolic content of some herbal teas. AB - The total antioxidant capacity of the aqueous extracts of some endemic herbs prepared as infusions by steeping these herbs in hot water--was assayed with bis(neocuproine)copper(II) chloride, also known as the cupric ion reducing antioxidant capacity (CUPRAC) reagent, which was easily accessible, rapid, stable and responsive to both hydrophilic and lipophilic antioxidants. The highest antioxidant capacities of some herbal teas available in the Turkish market were observed for scarlet pimpernel (Anagallis arvensis), sweet basil (Ocimum basilicum), green tea (Camellia sinensis) and lemon balm (Melissa officinalis), in this order (1.63, 1.18, 1.07, and 0.99 mmol trolox equivalent (TR)/g, respectively). For infusions prepared from ready-to-use tea bags, the CUPRAC values were highest for Ceylon blended ordinary tea (4.41), green tea with lemon (1.61), English breakfast ordinary tea (1.26) and green tea (0.94), all of which were manufactured types of C. sinensis. Following the strongest antioxidant herbs with capacities close to or slightly exceeding 1.0 mmol TR/g, sage, thyme, coriander, coltsfoot, blackberry and immortelle (Helichrysum) exhibited capacities around 0.5 mmol TR/g. The correlation of the Folin total phenolic content of herbal teas with their CUPRAC and ABTS (2,2'-azinobis(3 ethylbenzothiazoline-6-sulfonic acid) diammonium salt) total antioxidant capacities gave linear curves with correlation coefficients of 0.966 and 0.936, respectively, showing that the CUPRAC assay results better correlated with total phenolic content of herbal teas. Absorbance versus concentration data at different dilutions and upon standard additions of model antioxidant compounds (trolox and quercetin) to herbal tea infusions showed that the absorbances (at 450 nm of the CUPRAC method) due to different antioxidant compounds in herbal tea infusions were additive; that is, the tested antioxidants did not chemically interact to cause apparent deviations from Beer's law. PMID- 17135021 TI - Nutritional quality of leaves of some genotypes of mulberry (Morus alba). AB - This study was undertaken to determine the proximate composition, vitamins, minerals and the antinutritional factor tannic acid in leaves of six genotypes of mulberry. The results showed that in fresh mulberry leaves the proximate composition values ranged from 71.13 to 76.68% for moisture, from 4.72 to 9.96% for crude protein, from 4.26 to 5.32% for total ash, from 8.15 to 11.32% for Neutral Detergent Fiber (NDF), from 0.64 to 1.51% for crude fat, from 8.01 to 13.42% for carbohydrate and from 69 to 86 kcal/100 g for energy. In dried mulberry leaf powder, moisture ranged from 5.11 to 7.24%, crude protein from 15.31 to 30.91%, total ash from 14.59 to 17.24%, NDF from 27.60 to 36.66%, crude fat from 2.09 to 4.93%, carbohydrate from 9.70 to 29.64% and energy from 113 to 224 kcal/100 g. Among vitamins ascorbic acid and beta-carotene were found to range from 160 to 280 mg/100 g and from 10,000.00 to 14,688.00 microg/100 g, respectively, in fresh mulberry leaves and from 100 to 200 mg/100 g and from 8438.00 to 13,125.00 microg/100 g, respectively, in dried mulberry leaf powder. The minerals iron, zinc and calcium were observed in the ranges of 4.70-10.36 mg/100 g, 0.22-1.12 mg/100 g and 380-786 mg/100 g, respectively, for fresh mulberry leaves, and 19.00-35.72 mg/100 g, 0.72-3.65 mg/100 g and 786.66-2226.66 mg/100 g, respectively, for dried mulberry leaf powder. The tannic acid ranged from 0.04 to 0.08% in fresh leaves and from 0.13 to 0.36% in dried leaf powder. PMID- 17135022 TI - Coping strategies and nutritional health in rural Niger: recommendations for consumption of wild plant foods in the Sahel. AB - People who live in food and water deficit regions of Sahelien West Africa employ various coping strategies as they attempt to meet their food and water needs. In this paper we discuss various coping strategies employed by rural Nigeriens living in the Tanout and Mirriya administrative regions of central Niger. In rural Niger people often harvest or buy wild plant foods to eat. Laboratory studies of the nutritional content of these plants indicate that there are benefits to eating wild plant foods. In this study we summarize the results of field research conducted during the summer of 2002 on the use of wild plant foods in three regions of rural central Niger. Comparing local use of various wild plant foods with major nutrition-related health problems including protein deficiency, essential fatty acid deficiency, iron deficiency and iron deficiency anemia, calcium deficiency rickets, and zinc deficiency, suggests potential recommendations for consumption of these plants. However, further research on the bioavailability of these nutrients is needed to confirm the potential benefits of these plants. PMID- 17135023 TI - Estimated intakes of isoflavones and coumestrol in Korean population. AB - The dietary intakes and sources of isoflavones and coumestrol were estimated for each age group of Koreans based on data from the Korean Nutrition Survey conducted in 1998. For quantitative data on the levels of isoflavones and coumestrol, our previous study monitoring phytoestrogens in 220 Korean leguminous foodstuffs was employed and the median value for each food was adopted. The total isoflavones and coumestrol intake per capita was estimated as 23.3 mg/day, which constituted 14.2 mg daidzein, 6.7 mg genistein, 0.9 mg glycitein, 1.0 mg formononetin, 0.2 mg biochanin A, and 0.3 mg coumestrol. The top five foods arrowroot, soybean paste, tofu, soybean, and soybean sprout contributed to 88.2% of isoflavone intake, with the corresponding intake from each food being 8.3 mg/day, 4.9 mg/day, 2.6 mg/day, 2.5 mg/day, and 2.0 mg/day, respectively. Starting at age 3-6, the contributions of fermented soy products to the isoflavones intakes were around 30%. Soybean sprout was a major source of coumestrol intake in Koreans. Slight differences in the preference of these foods were observed among the various age groups. As regards the total isoflavone intakes, the highest value was 33.6 mg/day for people age 30-49, followed by age 50-64 (26.4 mg/day), 20-29 (21.0 mg/day), >or=65 (18.8 mg/day), 1-2 (14.5 mg/day), 7-12 (12.4 mg/day), 13-19 (10.1 mg/day), and 3-6 (8.9 mg/day). The intake levels are likely to be exceeded in groups who have preferably consumed high phytoestrogen-containing foods such as soy-protein-based infant formula and arrowroot. PMID- 17135024 TI - Characteristics of grains and oils of four different oats (Avena sativa L.) cultivars growing in Turkey. AB - Some physical and chemical properties of four oat (Avena sativa L.) varieties (BDMY-6, BDMY-7, Che-Chois and Y-2330) harvested from Konya in Turkey were investigated. The weight of the grain, moisture, crude protein, crude ash, crude fibre, crude energy, crude oil and water-soluble extract contents of all oat variety grains were analysed. Contents of aluminium, calcium, cadmium, phosphorus, magnesium, zinc, lead, potassium and manganese were also determined in the oat grains. The specific gravity, refractive index, free fatty acids, peroxide value, saponification number and unsaponifiable matter were determined in the grain oil. Tocopherol contents of these four oat grain oils were measured. Palmitic acid (15.72%), oleic acid (33.97-51.26%) and linoleic acid (22.80 35.90%) were found to be rich in protein, oil, fibre, unsaturated fatty acids and minerals, suggesting that they may be valuable for food uses. Due to high nutritive values, it is recommended to process for healthy food products. PMID- 17135025 TI - Honey ameliorates influence of hemorrhage and food restriction on renal and hepatic functions, and hematological and biochemical variables. AB - The objectives were to assess the effects of various diets, including total food restriction with 50% honey feeding, total food restriction with 50% dextrose feeding or adlibitum (control group) commercial regular diet, on the hematology and biochemical variables, and to assess the effects of the various diets on the influence of acute blood loss on the same parameters. Thirty Sprague-Dawley albino rats were divided into three groups, 10 rats each: group A, fed a commercial regular diet; group B, total food restriction with 50% dextrose feeding; and group C, total food restriction with 50% honey feeding. After 8 days of feeding, rats were subjected to acute blood loss (6 ml/kg) and blood investigations were performed. After acute blood loss, the same feedings were continued for a further 8 days and the blood tests were repeated at day 8 post bleeding. Total food restriction with 50% dextrose feeding compared with commercial regular diet reduces hematological and biochemical variables. Total food restriction with 50% honey feeding compared with total food restriction with 50% dextrose feeding causes a greater reduction in fasting blood glucose, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and triacylglycerol. Acute blood loss causes elevation of white blood cells, lymphocyte percentage, fasting blood sugar, blood urea nitrogen, alkaline phosphatase and triacylglycerol, and a reduction in serum albumen, protein, cholesterol, AST, serum creatinine and hemoglobin; the results are significant (P<0.05) concerning fasting blood glucose, AST, alkaline phosphatase, serum albumin and protein. A significant reduction in fasting blood glucose, white blood cells, BUN, AST, ALT, alkaline phosphatase and triacylglycerol, and a significant elevation of hemoglobin and serum albumin are obtained after acute blood loss in rats on total food restriction with 50% honey feeding as compared with the other two groups. Total food restriction with 50% honey feeding increases serum albumin, serum protein, fasting blood glucose, and causes lower reduction in hemoglobin as compared with the other groups. Conclusively, honey feeding during total food restriction significantly modifies and ameliorates biochemical and hematological changes observed after acute blood loss. This will pave the way to use honey as part of bleeding management and during a food restriction regimen. PMID- 17135026 TI - Consumer acceptability of low-sugar watermelon sweetened with non-calorie sweetener by a Native American community. AB - Watermelons are a good source of lycopene, a carotenoid that exhibits antioxidant activity and may protect against some cancers. However, intake of watermelon may be restricted for individuals who have diabetes or those who limit carbohydrate intake. A low-sugar watermelon was developed at Lane, Oklahoma using traditional plant breeding techniques. The objective of this study was to determine whether the artificially sweetened low-sugar watermelon was acceptable with Native Americans, a group with a high incidence of diabetes. The red flesh from a low sugar watermelon and a commercial variety of watermelon was removed and cut into cubes. Low and high levels of artificial sweetener were added to the low-sugar watermelon. Students at a Native American school (Grades 1-12) and adults at a Native American Feeding Center were asked to rate how much they liked or disliked the watermelon using a seven-point hedonic scale. Sugar composition, pH, lycopene and other carotenoids were analyzed from samples using established methods. The pH, lycopene, beta-carotene and total carotenoid levels were similar among fruit. Artificially sweetened fruit were rated slightly more acceptable in taste than the commercial control watermelons by both age groups. The low-sugar watermelons were lower in sugar composition but were comparable with conventional melons in all other quality factors and were found acceptable in taste by a broad age group of Native American consumers. PMID- 17135027 TI - 'Cyanidin volumetric index' and 'chromaticity coordinates ratio' to characterize red raspberry (Rubus idaeus). AB - The object of this work is presented in a larger research project concerning 'New indexes to evidence the nutritional quality of small fruits' in progress at the Analytical Food Research Laboratories, University of Milan. The present paper contains data that contribute to the analytical characterization of 12 varieties of red raspberry through the high-performance liquid chromatography determination of the aglycon 'cyanidin' derived from chemical hydrolysis of berries. Even more interesting results are the proposal of the 'cyanidin volumetric index', by which it is possible to compare different red raspberry varieties with higher meaningfulness. A new possible correlation between the ratio of chromaticity coordinates 'a/b' and the cyanidin content of red raspberries has been identified. PMID- 17135028 TI - The effect of soaking and cooking on the oxalate content of taro leaves. AB - Pacific Island people commonly eat taro (Colocasia esculenta var. Schott) as a staple food in their home islands and also like to consume this familiar food when living in New Zealand. Some of these foods are imported from the islands and some attempts are, currently, being made to grow these crops in New Zealand. The taro leaves in this experiment were grown in a greenhouse in the North Island of New Zealand. The soluble oxalate content of the raw leaves was 236.10 mg oxalate/100 g wet matter (WM). Soaking the raw leaves in water for 30 min marginally reduces the soluble oxalate content by leaching into the tap water. Soaking for 18 h results in a 26% reduction in the soluble oxalate content of the raw leaves. During the soaking treatments the insoluble oxalate (calcium oxalate) content of the leaves remained constant (mean 171.64 mg oxalate/100 g WM). Boiling the taro leaves resulted in a 36% loss of soluble oxalates, while the soluble oxalate content of baked tissue was very similar to the raw tissue. The mean insoluble oxalate content of the raw, boiled and baked tissue was 226.28 mg oxalate/100 g WM. Overall, boiling the taro leaves was an effective way of reducing the soluble oxalate content of the cooked tissue. PMID- 17135029 TI - Contribution to the study of camel milk fat globule membrane. AB - The camel milk fat globule membrane has been characterized according to several approaches. Compared with the cow milk fat globule membrane, various specificities have been revealed. Its physicochemical composition showed a poor content in proteins, and a higher content in neutral lipids and in phospholipids. The mechanical properties measured at low (4 degrees C, 20 degrees C) and high temperatures (40 degrees C, 45 degrees C and 50 degrees C) using a film balance are different when the camel milk fat globule membrane is spread at the air-water interface. The thermal study revealed an important proportion of high-melting triacylglycerols that involves fatty acids with long chains. PMID- 17135030 TI - Quercetin is bioavailable from a single ingestion of grape juice. AB - The in vivo bioactivity of polyphenols will depend on their bioavailability. Grape juice is an important source of dietary phenolics. This paper reports results that prove that quercetin (3,3',4',5,7-pentahydroxyflavone) is bioavailable after a single ingestion of red grape juice by healthy volunteers. Blood plasma samples were collected before and after 2 h of ingestion of 100 ml of concentrated grape juice (n = 14), and of a placebo solution (n =6). Significant differences in the variation of the total plasma quercetin content (before and after ingestion) between the grape juice ingestion group (3.1 microg/l increase, as a mean) and the placebo group (6.0 microg/l decrease, as a mean) were found. This relatively low increase in comparison with that obtained after 2 h of ingestion of onions (201 microg/l, as a mean) and with those reported in the literature for other foods/beverages was attributed to differences in the amount of quercetin ingested, in the form in which quercetin is present, and in the food matrix. PMID- 17135031 TI - Carotenoid and vitamin content of Karat and other Micronesian banana cultivars. AB - We previously found high carotenoid levels in Karat and other Micronesian bananas, indicating potential importance for alleviating vitamin A deficiency and other nutritionally related health problems in the Federated States of Micronesia. Past work focused on carotenoid and mineral analyses, whereas here we investigated 16 cultivars (most not previously analysed) for a broader micronutrient profile, including seven vitamins. Karat carotenoid levels were higher than in previous analyses, confirming Karat as exceptionally carotenoid rich. We identified an additional 10 carotenoid-rich cultivars, expanding the range having potential for alleviating vitamin A deficiency. A striking finding is the high riboflavin level in Karat, including high levels of uncharacterized flavonoids. Niacin and alpha-tocopherol are at levels that may contribute importantly to dietary intake within normal patterns of consumption. These data present a more complete basis for promoting the nutritional benefits of these banana cultivars where they are consumed in the Pacific, and potential benefits for promoting elsewhere. PMID- 17135032 TI - Use of oral contraceptives in infertile patients: A descriptive review. AB - In the past and currently, there are several applications of oral contraceptives (OCs) in the field of infertility. The aim of the present descriptive review is to analyze the old and new uses of OCs in infertile patients undergoing simple or multiple ovulation-induction programs. Available data seem to indicate that OCs given before treatment could be effective in patients who are resistant to clomiphene citrate and in poor responders. In addition, the use of OCs is useful for scheduling in vitro fertilization programs downregulated with gonadotropin releasing hormone-antagonists. PMID- 17135033 TI - Effect of the menstrual cycle and oral contraceptives on aromatase and cyclooxygenase-2 expression in adenomyosis. AB - OBJECTIVES: To determine whether aromatase expression in the eutopic endometrium and adenomyotic foci is affected by previous use of oral contraceptives containing gestodene, and to determine whether changes in cyclooxygenase-2 (COX 2) expression occur in adenomyosis during the menstrual cycle. PATIENT AND METHODS: This was a retrospective cohort study carried out in paraffin-embedded endometrial tissue obtained from patients with a histological diagnosis of adenomyosis obtained during the proliferative (n = 25) and luteal (n = 10) phases of the menstrual cycle and following the use of continuous oral contraception with gestodene/ethinyl estradiol (n = 7). COX-2 and aromatase expression were measured in both eutopic endometrium and adenomyotic foci using immunohistochemical methods. RESULTS: Aromatase expression was detected in 80% of the endometrial slices by immunohistochemistry. In positive cases, aromatase was mainly detected in the stromal cells of the eutopic endometrium, whereas in the adenomyotic foci this expression was negative in the majority of the cases. Oral contraceptives containing gestodene, on the other hand, were effective in suppressing aromatase expression in both eutopic and ectopic endometrium. COX-2 expression was detected by immunohistochemistry in the glandular epithelium of both eutopic endometrium and adenomyotic foci and there were no significant changes in its intensity throughout the menstrual cycle. CONCLUSION: Aromatase expression in the eutopic endometrium and adenomyotic foci is suppressed by oral contraceptives containing gestodene. Increased aromatase activity may be responsible for the persistent COX-2 expression during the luteal phase. PMID- 17135034 TI - Polymorphisms of the vitamin D receptor gene predict the onset of surgical menopause in Caucasian females. AB - We tested association of four single nucleotide polymorphisms (SNPs) of the vitamin D receptor gene (VDR) with age at surgical and natural menopause in a sample of Caucasians composed of 153 women with surgical and 260 with natural menopause. A significant association was observed between age at surgical menopause and two SNPs, rs1544410 (BsmI) and rs731236 (TaqI) (p < 0.05). For rs1544410, homozygotes of the minor allele, AA, had about two-fold higher risk of surgical menopause than homozygotes of the major allele, GG (95% confidence ratio (CI) 1.09-3.82). For rs731236, the CC subjects had a greater chance of surgical menopause than the TT subjects (odds ratio = 2.01, 95% CI 1.07-3.78). Since rs1544410 and rs731236 are in strong linkage disequilibrium, the haplotypes based on these two loci were also tested. The haplotype AC was highly significantly associated with age at surgical menopause (p = 0.008). Women with this haplotype had surgical menopause on average 2.8 years earlier than non-carriers. These results reveal the potential effect of the VDR gene on ovaries and uterus, and suggest that its SNPs can be used as predictors of genetic susceptibility for early surgical menopause and respective causal health problems. PMID- 17135035 TI - Effects of estradiol alone and combined with norethisterone acetate on pulse-wave velocity in hypertensive postmenopausal women. AB - BACKGROUND: Arterial hypertension and postmenopausal reduction of estrogen levels may be involved in modifications of the stiffness of large arteries. OBJECTIVES: To evaluate the pulse-wave velocity (PWV) and indirectly the arterial stiffness in hypertensive postmenopausal women submitted to hormone therapy with estradiol alone or combined with norethisterone acetate. SUBJECTS: Forty-five hypertensive postmenopausal women were double-blindly, randomly assigned to three arms of treatment: placebo (group I); estradiol 2 mg/day (group II); or estradiol 2 mg/day and norethisterone acetate 1 mg/day (group III). METHODS: Arterial stiffness was assessed from PWV measurements of the common carotid and femoral arteries (CF-PWV) and the common carotid and radial arteries (CR-PWV) obtained using the automatic Complior(R) device, taken at baseline and after 12 weeks of treatment. RESULTS: After the 12-week treatment, values of CF-PWV and CR-PWV were not significantly different (p = 0.910 and p = 0.736, respectively) among the groups. Systolic blood pressure showed a positive correlation with CF-PWV in groups II and III (p = 0.02 and p < 0.001, respectively). CONCLUSIONS: PWV and arterial stiffness in postmenopausal hypertensive women did not reduce over a 12 week treatment with estradiol alone compared with the same period of treatment with estradiol combined with norethisterone acetate. PMID- 17135036 TI - Hormone replacement therapy in menopausal women: Past problems and future possibilities. AB - Oral administration of conjugated equine estrogens (CEE) with and without the synthetic progestin medroxyprogesterone acetate (MPA) in postmenopausal women is associated with side-effects that include increased risk of stroke and breast cancer. The current evidence that transdermal administration of estradiol may provide a safer alternative to orally administered CEE is reviewed. Transdermally administered estradiol has been shown to be an efficacious treatment for hot flushes possibly without the increase in blood clotting that is associated with administration of oral CEE. Further, natural progesterone may have a more beneficial spectrum of physiological effects than synthetic progestins. The substantial differences between CEE compared with estradiol and estriol, as well as the differences between synthetic MPA and natural progesterone, are detailed. Estriol is an increasingly popular alternative hormone therapy used for menopausal symptoms. There is evidence that estriol, by binding preferentially to estrogen receptor-beta, may inhibit some of the unwanted effects of estradiol. New clinical trials are needed to evaluate the safety and efficacy of topically or transdermally administered combinations of estradiol, estriol and progesterone. Future studies should focus on relatively young women who begin estrogen supplement use near the start of menopause. PMID- 17135037 TI - In vitro estrogenic activity of formononetin by two bioassay systems. AB - AIM: To study the estrogenic activity of formononetin in vitro. METHODS: We have established a highly sensitive bioassay system by placing estrogen-responsive elements upstream of the luciferase reporter gene, and used this assay to determine the estrogenic activity of formononetin. Cell growth was measured by the MTT (3-(4,5-dimethylthioazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and MG-63 cell function was studied by measuring alkaline phosphatase activity. RESULTS: Formononetin activated expression of the estrogen-responsive reporter gene in human breast cell line MCF-7 in a concentration-dependent manner (0.5-500 microM), and this activation was inhibited by estrogen antagonist (ICI 182780 at 100 nM). Furthermore, it induced the proliferation of MCF-7 breast cancer cells and MG-63 osteosarcoma cells, and it also increased the alkaline phosphatase activity in MG-63 cells. CONCLUSION: Formononetin is a phytoestrogen that exhibits variable degrees of estrogen receptor agonism in different test systems. PMID- 17135038 TI - Relationship between pregnancy outcomes and maternal vitamin D and calcium intake: A cross-sectional study. AB - Poor maternal vitamin D status affects fetal and infant skeletal growth. The aim of the present study was to determine the association between newborn outcomes and maternal calcium and vitamin D intakes. Four hundred and forty-nine pregnant women, healthy at the point of delivery, and their newborns were enrolled in the study, which was performed in three university hospitals in Tehran in March 2004. Maternal anthropometric data and energy, protein, calcium and vitamin D intakes were collected, and newborn outcomes (weight, length, head circumference and 1 min Apgar score) were determined. Almost two-thirds of the mothers (64.3%) took no supplements during pregnancy. Only one-third of the mothers (33.8%) had adequate intakes of calcium and vitamin D (from supplements and foods) compared with the Recommended Dietary Allowances. Mean length at birth and 1-min Apgar score were higher in newborns whose mothers had adequate calcium and vitamin D intake than in newborns whose mothers had inadequate intake (p = 0.03 and p = 0.04, respectively). Significant correlations were found between adequate maternal calcium and vitamin D intake and both appropriate birth weight and 1-min Apgar score of newborns and weight gain of mothers during pregnancy. Informing mothers of the critical importance of consuming adequate amounts of calcium and vitamin D seems necessary. PMID- 17135039 TI - Insulin resistance and body composition in Turner syndrome: Effect of sequential change in the route of estrogen administration. AB - The aim of the present study was to examine the impact of sequential change in the route of estrogen administration on body composition and insulin resistance in patients with Turner syndrome (TS) using cyclical hormone replacement therapy (HRT) with conjugated equine estrogens (CEE) plus medroxyprogesterone acetate (MPA). We carried out a self-controlled study of nine non-obese patients with TS, with an average age of 23 +/- 4.9 years. Body mass index (BMI), waist-to-hip ratio (WHR), fasting glycemia, insulin tolerance (glucose disappearance constant during an insulin tolerance test, kITT) and body composition (dual-energy X-ray absorptiometry) were studied after 1 year's use of CEE plus MPA and repeated after 1 year's use of 17beta-estradiol gel with the same schedule of MPA administration. We did not observe any difference between the oral and percutaneous HRT with regard to BMI, WHR and insulin tolerance (kITT: 4.9 +/- 1.5 vs. 5.3 +/- 1.5%/min, p = 0.8). During administration of the 17beta-estradiol gel a tendency to increased total lean mass (p = 0.054) was observed. We conclude that sequential change in the route of estrogen administration in TS patients using cyclical HRT with CEE and MPA does not affect insulin resistance, although use of percutaneous 17beta-estradiol gel seems to exert favorable changes in body composition. PMID- 17135040 TI - Thiophenes from Echinops latifolius. AB - Six thiophenes were isolated and purified from ethanol extract of the roots of Echinops latifolius Tausch. Their structures were identified on the basis of spectral data. Among them, 5-(3-hydroxmethyl-3-isovaleroyloxyprop-1-ynyl)-2,2' bithiophene (6) is a new compound, and 5-(3-hydroxy-4-isovaleroyloxybut-1-ynyl) 2,2'-bithiophene (5) was isolated from this plant for the first time. PMID- 17135041 TI - Two new terpenoid glucosides from Aster flaccidus. AB - Two new terpenoid glucosides, namely 2-O-beta-D-glucopyranoside-vicodiol (1) and 10-O-beta-D-glucopyranoside-oplopanone (2), along with seven known compounds, were isolated from the aerial part of Aster flaccidus (composite), a traditional Chinese herb medicine. The structures of 1 and 2 were established by spectroscopic methods, especially 2D NMR experiments. PMID- 17135042 TI - A new chromone from Cassia nodosa. AB - A new chromone named as 5,4'-dihydroxy-7-methyl 3-benzyl chromone (1) along with three known flavonoid compounds as unsubstituted flavone, kaempferol-3-o rhamnoside and quercetin-3-o-arabinoside have been isolated from the leaves of Cassia nodosa. Their structures have been established by means of chemical and spectral evidences (IR, UV, 1H-NMR, 13C-NMR and mass spectra). PMID- 17135043 TI - A new labdane diterpene from Leonurus heterophyllus. AB - A new labdane diterpene, heteronone B (1), together with a known labdane diterpene, heteronone A (2), have been isolated from the aerial part of Leonurus heterophyllus. Their structures were established mainly by 1D and 2D NMR analysis and the stereochemistry of 2 was confirmed by single-crystal X-ray diffraction analysis. PMID- 17135044 TI - Ginsenine, a new alkaloid from the berry of Panax ginseng C. A. Meyer. AB - A new indole alkaloid, ginsenine, with a seven-membered lactam unit, was isolated from the berry of Panax ginseng. Its structure was established on the basis of extensive NMR (1H- and 13C-NMR, 1H-1H COSY, DEPT, HMQC, HMBC), IR, and ESI-MS analysis. PMID- 17135046 TI - Two new cardenolides from the roots of Streptocaulon griffithii. AB - Two new cardenolides, 3beta,5beta,14beta-trihydroxyl-card-16,20(22)-dienolide (1) and 3-O-beta-D-glucopyranosyl-5beta,14beta-dihydroxyl-card-16,20(22)-dienolide (2), together with seven known compounds identified as digitogenin (3), 16-O acetylgitoxigenin (4), periplogenin (5), 16-O-acetylperiplogenin (6), periplogenin digitoxoside (7), periplogenin-3-O-beta-D-glucopyranoside (8) and periplogenin-3-O-beta-D-glucopyranosyl-(1-->4)-O-beta-D-digitoxopyranoside (9) were isolated from the roots of Streptocaulon griffithii. Their structures were elucidated on the basis of spectroscopic data. PMID- 17135045 TI - A new chloro-monoterpene from the mangrove endophytic fungus Tryblidiopycnis sp. (4275). AB - A new chloro-monoterpene (compound 1) and three known compounds, peroxyergosterol, uracil and methylisocoumarin, were isolated from the ethyl acetate extract of the fermentation broth of the mangrove endophytic fungus Tryblidiopycnis sp. (4275) obtained from Kandelia woody tissue from Mai Po, Hong Kong. Its structure was determined spectroscopically and by X-ray crystallographic analysis. PMID- 17135047 TI - Structure of lasiansine from Aconitumnagarum var. lasiandrum. AB - A new C19-diterpenoid alkaloid, lasiansine (1), was isolated from the roots of Aconitu nagarum var. lasiandrum (Ranunculaceae) together with six known diterpenoid alkaloids. The structure of 1 was elucidated by spectral methods (1H NMR, 13C-NMR, 2D-NMR, HRMS, IR), and the 13C-NMR spectrum of 16-epipyroaconine (3) and the single-crystal X-ray analysis of its derivative (5) are reported for the first time. PMID- 17135048 TI - Water-soluble constituents from aerial roots of Ficus microcarpa. AB - Three new water-soluble constituents [ficuscarpanoside B (1), (7E,9Z) dihydrophaseic acid 3-O-beta-D-glucopyranoside (4) and ficuscarpanic acid (6)] and the natural product 2,2'-dihydroxyl ether (7) have been isolated, together with three known compounds [(7S,8R)-syringoylglycerol (2), (7S,8R) syringoylglycerol-7-O-beta-D-glucopyranoside (3) and icariside D2 (5)] from the aerial roots of Ficus microcarpa. Identification of their structures was achieved by 1D and 2D NMR experiments, including 1H-1H COSY, NOESY, HMQC and HMBC methods and FAB mass spectral data. PMID- 17135049 TI - A new triterpene glycoside from Asterias rollentoni. AB - A new triterpene glycoside, rollentoside A, has been isolated from Asterias rollentoni Bell and identified as 3beta-O-{3-O-methyl-beta-D-xylopyranosyl-(1- >3)-O-beta-D-glucopyranosyl-(1-->4)-O-beta-D-quinopyranosyl-(1-->2)-O-beta-D xylopyranosyl}-16-beta-acetoxy-23S-acetoxy-holost-7-ene (1), together with a new natural product, rollentoside B (2). The structures of compounds1 and 2 were elucidated by extensive 1D and 2D NMR investigation (1H-1H COSY, TOCSY, HSQC, HMBC, NOESY). PMID- 17135050 TI - Diarylheptanoids from Alpinia officinarum. AB - A new diarylheptanoid, along with five known diarylheptanoids, was isolated from the rhizomes of Alpinia officinarum (Zingiberaceae). The structure of the new compound was determined to be trans,trans-1(3'-methoxy-4'-hydroxyphenyl)-7-phenyl 5-ol-4,6-dien-3-heptanone on the basis of spectral and chemical evidence. PMID- 17135051 TI - Heteroclitin H, a new lignan from Kadsura heteroclita. AB - A new dibenzocyclooctadiene lignan, heteroclitin H (1), was isolated together with seven known compounds, heteroclitin D (2), interiorin B (3), interiorin (4), neokasuranin (5), interiotherin C (6), gomisin J (7) and (+)-anwulignan (8), from the stems of Kadsura heteroclita. Their structures were elucidated by spectroscopic methods. This is the first report of the isolation of compounds 3, 5, 6, 7 and 8 from K. heteroclita. PMID- 17135052 TI - Effect of 17beta-oestradiol and ginsenoside on osteoporosis in ovariectomised rats. AB - To study the anti-osteoporosis effects and mechanism of action of oestradiol (E2) and ginsenoside (tR), we measured the bone mineral densities (BMD) of lumbar vertebra and tibia and analysed the tibia histological morphological data, as well observed the activity and the number of osteoblasts and the activity of alkaline phosphatase (ALP) and the concentration of cAMP. Results showed that E2 (400 microg kg- 1 week- 1) and tR (10, 20, 30 mg kg- 1 day- 1) were able to countervail the decreasing in BMDs of lumbar vertebra and tibia induced by OVX in rats (P<0.05); E2 (0.1 micromol l- 1) and ginsenoside Rg1 (1 micromol l- 1 and 10 micromol l- 1) were able to increase the number of osteoblasts, the activity of ALP and the concentration of intercellular cAMP in cultured osteoblast cells. The present findings suggest that E2 and tR have an anti-osteoporosis effect in ovariectomised rats. PMID- 17135053 TI - Two new 9,10-seco-cycloartanes from the seeds of Sphaerophysa salsula. AB - Two new 9,10-seco-cycloartanes, named sphaerophyside SC (1) and sphaerophyside SD (2), together with four known compounds (3-6), were obtained from the ethanol extract of the seeds of Sphaerophysa salsula. The structures of these compounds were elucidated on the basis of spectral and chemical evidences. Compounds 3-6 were isolated from the plant for the first time. PMID- 17135054 TI - Chromone glycosides from Knoxia corymbosa. AB - Four new chromone glycosides, corymbosins K1-K4 (3-6), together with two known compounds, noreugenin (1) and undulatoside A (2), were isolated from the whole plant of Knoxiacorymbosa (Rubiaceae). The structures of the new compounds were established through extensive NMR or X-ray spectroscopic analysis as 7-O-beta-D allopyranosyl-5-hydroxy-2-methylchromone (corymbosin K1, 3), 7-O-beta-D-6 acetylglucopyranosyl-5-hydroxy-2-methylchromone (corymbosin K2, 4), 7-O-[6-O-(4-O trans-caffeoyl-beta-D-allopyranosyl)]-beta-D-glucopyranosyl-5-hydroxy-2 methylchromone (corymbosin K3, 5) and 7-O-[6-O-(4-O-trans-feruloyl-beta-D allopyranosyl)]-beta-D-glucopyranosyl-5-hydroxy-2- methylchromone (corymbosin K4, 6). Compounds 2-5 were subjected to test their immunomodulatory activity invitro. PMID- 17135055 TI - Pemphigus in the XXI century: new life to an old story. AB - In this new century of pemphigus research, the search for novel treatments is switching from a monospecific approach, focused on immunosuppression, to a polyspecific approach that includes drugs acting on novel pathophysiologic pathways. Current research argues that acantholysis in pemphigus occurs as an active process resulting from intracellular signaling triggered as a result of IgG binding to the keratinocyte membrane antigens in a receptor-ligand fashion. Recent progress regarding the pathophysiology of pemphigus acantholysis led to, or was accompanied by, breakthrough discoveries of safer treatments. Both the identification of cell-surface receptors to acetylcholine among the nondesmoglein (Dsg) targets for pemphigus antibodies, and the elucidation of the cholinergic control of keratinocyte cell adhesion provide an explanation for the therapeutic efficacy of cholinomimetics in patients with pemphigus. In patients' skin, Fas-L, TNFalpha, and, probably, IL-1alpha act as autocrine/paracrine co-factors for anti keratinocyte IgG. Thus, it appears that an array of interconnected signaling cascades is responsible for acantholysis and cell death in pemphigus. Future studies should define the signaling pathways mediating acantholysis that occur in individual pemphigus patients and identify the membrane proteins (receptors) triggering signaling along a specific pathway upon their ligation by autoantibodies. It will be important to determine which pathway 1) leads directly to a loss of cell-cell adhesion (primary pathway), 2) which is being activated due to cell shrinkage/detachment (secondary pathway), 3) which contributes to utilization of altered proteins and organelles (scavenging pathway), and 4) which represents the cell defense (protective pathway). To dissect out the signaling pathways originating from binding of pemphigus IgG to non-Dsg targets on the keratinocyte plasma membrane experiments should be performed in cultures of murine keratinocytes grown from the Dsg3-/- mice or human keratinocytes with the knocked-down expression of the Dsg1 and/or Dsg3 gene by the RNA interference. PMID- 17135056 TI - Interactive analysis of Belgian vital statistics on the Internet. AB - The purpose of the Centre for Operational Research in Public Health (CORPH) is to optimize the accessibility to health information, thus making it possible to measure and follow up the health status of the Belgian population. The Standardized Procedures for Mortality Analysis (SPMA) software was developed in order to facilitate the use of vital statistics for health policy-makers and scientific researchers. Nowadays, SPMA is available on the Internet, because accessibility to health information is crucial. SPMA serves via a system of menus as the interface between databases (population, birth, and mortality) on one hand and statistical procedures on the other hand. Users can choose the parameters such as year, cause of death, geographical level, and statistical indicator, and so dynamic reports are produced 'on demand'. These procedures are available for the following modules: overall mortality, specific cause mortality, and perinatal statistics. Analysis can be carried out for one specific year or for a period over time. Pre-defined procedures accessible through menus make SPMA user friendly, as it can be used without any preliminary knowledge of the statistical package. Tables, charts, or maps display the results. Users need only an Internet browser to access the application. PMID- 17135057 TI - Changing availability and cost of Internet physician consultations and prescription medications. AB - INTRODUCTION: Internet use by the public to obtain medical information and services continues to grow. In 1999, we found cost of general physician visits was 15% higher, and cost of medications was 10% greater via the Internet than from community providers. The goal of this study was to re-examine changes in product availability and costs to consumers four years later. DESIGN: We searched multiple websites 1 June 2003 - 31 August 2004, offering physician consultations and prescription medications. We compared mean cost of Internet physician visits and price per pill to costs of community-based general physicians and pharmacies in the Philadelphia region. RESULTS: We found 144 sites worldwide providing physician services and prescription medications. Mean charge for an Internet physician consultation was 55 dollars, 8.3% lower than for a comparable visit to a community practitioner. Mean cost per pill across 204 available medications was 36.7% higher at Internet sites than at local pharmacies. Shipping and handling (S&H) added a mean of 15.40 dollars. Total cost to Internet consumers was similar whether or not Internet sites charged for a physician consultation and/or S&H. Obtaining physician visits and medications over the Internet was about 40% more costly than in the local community. CONCLUSION: The Internet continues to hold great promise for provision of health and medical care services by expanding access. However, increased access to physician care and medications entails higher cost, and quality of physician services and pharmaceuticals provided remain controversial. PMID- 17135058 TI - A framework for comparing video methods used to assess the clinical consultation: a qualitative study. AB - BACKGROUND: Single-channel video is an established method for assessing clinical consultation in training general practitioners; however, it is hard to infer the body language of the doctor or how information in the consultation is being integrated into the medical record. A three-channel video was developed combining the conventional view with a camera looking at the doctor's facial expression and copying the video output from the monitor. However, the choice of three channels and camera angles selected has not been critically appraised. OBJECTIVE: To develop criteria for comparing single and multi-channel approaches to video recording of the consultation. METHODS: Single channel and three-channel recordings of simulated consultations were shown to a panel of 12 health professionals and interviews were conducted to gather their opinions on the level of information presented, quality and assessment. The transcripts were analysed thematically. RESULTS: It was found that in providing visual information the three-channel video was superior to the single channel video. The major elements needed for comparison of the two techniques would be the ability of the video to pick up quantifiable non-verbal communication of the doctor and the patient, and the ability to qualitatively and quantitatively reflect the use and impact of the computer on the consultation. The information provided by the three-channel video could be further classified to essential, desirable and redundant to guide the future development of the multi-channel video. CONCLUSIONS: Multi-channel methods should be able to capture the following information: body language and facial expression of doctor and patient; and how the doctor's knowledge and information collected in the consultation are synthesized into the medical record. PMID- 17135059 TI - Internet health-information seekers in Croatia--who, what, and why? AB - The aim of this study is to investigate the characteristics of the Internet users in Croatia related to seeking health information, which information they seek, the reasons, and the outcomes of that information on them. We surveyed the users of the Internet health portal 'Cybermed', in spring 2003 and autumn 2004. Responses were collected from a pop-up questionnaire, which appeared at every website visit. There were 369 respondents in 2003 and 580 respondents in 2004. Most respondents were women (82% in 2003 and 80% in 2004). While more than half of the respondents (55%) in 2003 had a college or university degree, this changed in 2004, when only 40% of respondents had a college or university degree (P < 0.001). Also, we recorded a significant age-related change, suggesting that the average age of the respondents increased in 2004 (P < 0.001). The leading motivation for seeking medical information was 'unanswered questions after visiting a physician'. Most respondents (90% in 2003, 87% in 2004) went online to seek information on a specific illness or condition. Women sought information for friends and colleagues more often than men (42% vs. 28%, P = 0.031) in 2003, while these differences were diluted in 2004. Approximately half of all respondents discussed the information they found online with their physicians. Patients who feel they were given insufficient information, or simply seek more information than the physician provided, turn to using online health information, which no longer seems to be reserved to those with the highest education degrees. Health-care providers should consider creating health-related Internet portals, supplementing the information they provide. PMID- 17135060 TI - Hypnotizability and somatic complaints: a gender-specific phenomenon. AB - The relationship between hypnotizability and somatic illness was measured in 45 college students. Several weeks after completing the Waterloo-Stanford Group C Scale (WSGC), participants filled out a somatic-complaint checklist and measures of psychopathology. Results indicated a positive correlation between hypnotizability and somatic illness, and the relationship was stronger for female participants. In contrast to the quadratic model proposed by Wickramasekera, the current data demonstrated a linear relationship between hypnotizability and somatic complaint. Further analyses showed that somatic complaints were associated with hallucination and imagery items, corresponding to the perceptual cognitive factor identified in Woody, Barnier, and McConkey's (2005) factor analysis of the Stanford Hypnotic Susceptibility Scale, Form C. The results call into question some claims that high hypnotizability is an adaptive and healthy trait. PMID- 17135061 TI - Fractal analysis of EEG in hypnosis and its relationship with hypnotizability. AB - Fractal analysis was applied to study the trends of EEG signals in the hypnotic condition. The subjects were 19 psychiatric outpatients. Hypnotizability was measured with the Hypnotic Induction Profile (HIP). Fifty-four sets of EEG data were analyzed by detrended fluctuation analysis (DFA), a well-established fractal analysis technique. The scaling exponents, which are the results of fractal analysis, are reduced toward white noise during the hypnotic condition, which differentiates the hypnotic condition from the waking condition. Further, the decrease in the scaling exponents during hypnosis was solely associated with the eye-roll sign within specific cortical areas (F3, C4, and O1/2) closely related to eye movements and attention. In conclusion, the present study has found that the application of the fractal analysis technique can demonstrate the electrophysiological correlations with hypnotic influence on cerebral activity. PMID- 17135062 TI - Hypnotic depth and response to suggestion under standardized conditions and during FMRI scanning. AB - Hypnosis is a potentially valuable cognitive tool for neuroimaging studies. However, understandable concern that Magnetic Resonance Imaging (MRI) in particular may adversely affect hypnotic procedures remains. Measurements of hypnotic depth and responsiveness to suggestions were taken using a standardized procedure that met all the requirements for functional MRI (fMRI). Testing outside the scanning environment showed reliable and stable changes in subjective hypnotic depth, with no carryover once the hypnosis had been terminated. Within subject comparisons showed that the magnitude and pattern of these changes and the degree of responsiveness to hypnotic suggestion were not discernibly affected by the fMRI environment. It is concluded that hypnosis can be employed as a discrete and reliable cognitive tool within fMRI neuroimaging settings. PMID- 17135063 TI - A spanish version of the barber suggestibility scale for the puerto rican population. AB - Currently in Puerto Rico, there are no reliable and valid instruments to assess hypnotic responsiveness. The most widely utilized scales have not been scientifically translated and adapted with Puerto Ricans. In the present study, the Barber Suggestibility Scale (BSS) was translated and adapted using back translation and decentralization. The translated BSS (the ESB) was individually administered to Puerto Rican college students (N = 85). No significant differences were found among the mean scores for the current sample on the ESB and the 1965 sample on the BSS. Both samples had similar score distributions. The internal consistency of the ESB was adequate, and there were significant correlations among scale items and total scores. The authors conclude that the ESB is an adequate instrument to measure hypnotic response within the Puerto Rican population. PMID- 17135064 TI - Effectiveness of hypnosis in reducing mild essential hypertension: a one-year follow-up. AB - The present study investigates the effectiveness of hypnosis in reducing mild essential hypertension. Thirty participants were randomly assigned to hypnosis (standardized, individual 8-session hypnosis treatment) or to a control group (no treatment). Results show that hypnosis is effective in reducing blood pressure in the short term but also in the middle and long terms. We did not find any relationship between the practice of self-hypnosis and the evolution of blood pressure or between anxiety, personality factors, and therapeutic results. The implications of the results of the psychological treatment of hypertension are discussed. PMID- 17135066 TI - Evidence to support a positive transition into adulthood for youth with disabilities. PMID- 17135065 TI - Hypnotizability, eating behaviors, attitudes, and concerns: a literature survey. AB - The literature suggests that aspects of hypnotizability may be involved in the etiology and maintenance of self-defeating eating. However, interpretation of the published research findings has been complicated by the use of instruments that appear to have measured different or, at best, only related facets of the underlying constructs. This article reports relationships between weight, shape, dietary concerns, hypnotizability, dissociative capacity, and fantasy proneness. Implications for a key role for hypnosis in the treatment of eating behaviors, attitudes, and concerns are discussed. PMID- 17135067 TI - A critical appraisal of literature reviews about the transition to adulthood for youth with disabilities. AB - A critical appraisal of five review articles on the transition to adulthood for youth with disabilities was conducted to identify evidence about (1) the factors that help or hinder the transition process, and (2) "what's working" in transition services. The appraisal identified a number of important "success" factors and elements of service delivery that are worthy of consideration by service providers and researchers. These include the need for skill development of youth with disabilities, environmental supports, and an individualized approach to service delivery. All of the reviews identified the need for more evidence to support the implementation and evaluation of best practice models/approaches that address the complex issue of the transition from paediatric to adult services for youth with disabilities. PMID- 17135068 TI - The transition study: a look at youth and adults with cerebral palsy, spina bifida and acquired brain injury. AB - A group of 100 adults and 190 youth who have cerebral palsy (CP), spina bifida (SB), and acquired brain injuries from childhood (ABIc) participated in a multi method study focused on the transition to adult health care. The results show that 95% of youth and 61% of adults were living with their parents; 23% of the youth and 55% of adults were employed; and 60% of youth and 42% of adults reported "excellent" or "very good" health. The lowest health scores were reported by adults with SB. These findings provide a starting point for examining health issues specific to youth and young adults with CP, SB, and ABIc. PMID- 17135069 TI - Growing up ready: a shared management approach. AB - In order to help youth with physical disabilities and their families to plan for the transition to adulthood, well-planned service delivery is essential. This paper provides an account of the work of a children's rehabilitation centre to develop a transition framework reflecting evidence-based practice. Examination of current transition practices, a review of the literature, and site visits to health care facilities and universities were conducted to identify promising practices in the field of transition to adult services. A transition framework was designed to facilitate the adoption of a shared management approach for helping families and their children to grow up ready. Key elements of the transition framework are described and future plans discussed. PMID- 17135070 TI - An evaluation of the "Youth en Route" program. AB - Youth En Route (YER) is a transition program for youth and young adults with multiple disabilities. It offers a multifaceted approach that includes self discovery, skill development, and community experience. Underlying the service delivery model is a philosophy of self-determination. This program evaluation measured the self-determination skills, sense of personal control over life choices, and community participation of 34 youth prior to and one year following their involvement with YER. Youth reported statistically and clinically significant improvement from pretest to posttest with respect to both self determination and sense of personal control. Moreover, youth reported spending significantly more time at posttest than at pretest engaged in volunteer/work activities and community leisure activities. On average, youth reported high satisfaction with YER services. Practical and research implications are discussed. PMID- 17135071 TI - Creating connections: a community capacity-building project with parents and youth with disabilities in transition to adulthood. AB - The transition to adulthood presents many challenges for youth with disabilities and their families. Barriers in the environment often limit the full inclusion of these youth in daily community life. The purpose of this paper is to describe a community capacity-building (CCB) approach to facilitating the transition to adulthood for youth with developmental disabilities and their families. A pilot project that used a CCB approach with this population in one community in southcentral Ontario is described. The results of a qualitative, participatory evaluation demonstrate the benefits and challenges of this approach, with themes of increased community connections for youth and a greater awareness of their strengths and capacities. The perceived outcomes of the participants and the "lessons learned" for future initiatives using a CCB approach with different populations are discussed, as well as the fit between community capacity-building and occupational therapy. This pilot project demonstrates that a CCB approach has the potential to assist youth with disabilities to participate within their own communities. PMID- 17135072 TI - Caught in the web. PMID- 17135085 TI - Women's health: new frontier in advocacy & social justice research-introduction. PMID- 17135086 TI - Advocacy as a means to an end: assisting refugee women to take control of their reproductive health needs. AB - Negotiating reproductive rights is particularly complex for resettling migrant women from refugee backgrounds. In our ongoing work with women who have fled from countries in Africa and the Middle East, and have resettled in Australia and New Zealand, subtleties of discrimination and perceptions of human rights discriminations were revealed through the complex interplay between research and advocacy. Community Based Participatory Research (CBPR) has therefore been critical in assisting women to identify their needs and negotiate acceptable solutions with health services. This paper presents qualitative and quantitative findings of research with women from refugee backgrounds in Australia (n = 255) and New Zealand (n = 64). The research questions were a combination of community driven and researcher initiated issues and the projects developed through a continuous iterative process involving feedback from women in the community. We highlight the essential role of advocacy in CBPR and how that can enhance research quality. We argue for the justification of this approach as not only valid and credible but essential in research with these and other communities. PMID- 17135087 TI - Typologies of rural lay-health advocacy among rural women in Australia. AB - BACKGROUND: Public health advocacy effects changes in health behaviors and outcomes through applying health promotion expertise to specific groups. Advocacy occurs through the provision of tools to empower those who are either experiencing, or at risk of, a particular health status. RESEARCH OBJECTIVES: Health-care experiences of women living in rural Victoria, Australia, were explored in the context of generally poor access to reproductive health services. Women's experiences are investigated within the theoretical framework of lay health advocacy, i.e. relying on individual health care experience and knowledge to promote and improve the health care of others. METHODOLOGY: The study applied a qualitative design, and a self-identified sample of women was recruited through network sampling techniques. Fifty-seven women participated in in-depth interviews. RESULTS: Three types of lay-health advocacy emerged. Advocacy-seekers expected the researchers to use their experiences of poor health-care to educate health professionals to provide better quality care. Advocacy providers used their knowledge and experiences to take an active part in promoting the health care of other women. Story-tellers expected their narratives to empower other women or unidentified social groups to feel less isolated in their health care experiences. DISCUSSION: In providing narratives of their health-care, women were critical of social inequalities facing people living in rural Australia. Lay health advocates offer a cost-effective and appropriate option for reducing adverse health outcomes within resource-poor settings. Informed by women's narratives, we suggest strategies to enhance rural women's health-care. PMID- 17135088 TI - The culture of domestic violence advocacy: values of equality/behaviors of control. AB - Domestic violence advocacy is a culture unto itself. The themes it addresses, in combination with the dominant frames of discourse (Fairclough 1989) used in the daily conversations of advocates and their supporters, contribute to the stagnation of domestic violence advocacy as a profession and stunt its ability to address the non-homogenous, culturally diverse group of men, women and children living under the tyranny of violence within their homes. Social and political ties to funding institutions and government agencies reify existing concepts about domestic violence education and allow little opportunity for improvement. This article details the methodology and findings of a qualitative, ethnographic research project conducted at an undisclosed domestic violence agency. Qualitative results were analyzed using methods of grounded theory, discourse analysis and narrative analysis. Results revealed four major discourse themes that contribute to the current culture of domestic violence advocacy: (1) systemic contributions to the normalization of crisis, (2) cultural contradictions between paradigms and practice of domestic violence advocacy (3) ambiguity over the line between childhood and adulthood and (4) assumptions about "victim-hood". PMID- 17135089 TI - Cervical cancer services for Indigenous women: advocacy, community-based research and policy change in Australia. AB - Collaborative research undertaken in the state of Queensland, Australia, resulted in major changes in cervical cancer screening and treatment for Indigenous women. Guided by an Indigenous statewide reference group and with an Indigenous researcher playing a lead role, qualitative data were collected using interviews, focus groups, larger community meetings; and case studies were conducted with health workers and community members from diverse rural, remote and urban communities, to explore the different cultural and structural factors affecting understanding and awareness of cervical cancer and Indigenous women's use of and access to health services for screening, diagnosis and treatment. These data were supplemented by an analysis of clinical data and health service checklists. We discuss the methodology and summarize the key social and structural factors that discourage women from presenting for screening or returning for follow-up. These include women's misunderstanding of cervical cancer screening, fear of cancer, and distrust of health services, poor recall and follow-up systems, and the economic and social burden to women to presenting for treatment. We describe how the research process and subsequent activities provided Indigenous women with a vehicle for their own advocacy, resulting in important policy and program changes. PMID- 17135090 TI - Using participatory research to build an effective type 2 diabetes intervention: the process of advocacy among female Hispanic farmworkers and their families in Southeast Idaho. AB - The Formando Nuestro Futuro/Shaping our Future project (herewith, Formando) is a community-based participative research (CBPR) focused on type 2 diabetes. It was conceptualized and designed by a team of university-based researchers and community health workers (promotores). This article describes the process of establishing a CBPR project such as Formando and the most current results from that project. The Formando project is an example of health-focused advocacy with the Mexican agricultural workers in Southeast (SE) Idaho. To date, 172 qualitative interviews on participants' knowledge about type 2 diabetes have been carried out with farmworker women and their families. Biometric data (heights, weights, blood pressures and fasting blood glucoses) were obtained from participants. Fieldnotes, focus group discussions and key informants were used to triangulate findings. Significant quantitative findings include that age was significantly associated with Body Mass Index (BMI) (p < 0.001, Spearman Correlation < 0.001) and with elevated fasting blood glucose (p < 0.001, Spearman Correlation < 0.001). The qualitative interviews were thematically analyzed. Key themes associated with type 2 diabetes in this community were the connection between thinness and vanity, dieting and starvation and the onset of diabetes as a result of, what social scientists call, structural violence within the immigrants' daily lives. We conclude that long-term commitment to using the CBPR approach in these Mexican agricultural communities is an effective way to engage in health research and to establish real and meaningful dialogue with community members. PMID- 17135091 TI - Family and aging policy. PMID- 17135092 TI - Policies and practices in support of family caregivers - filial obligations redefined in Sweden. AB - This article provides an overview of how the expression of filial obligations has shifted over time in Sweden. Historically, and currently in many countries, the family, next of kin, and the social network are the only or major sources of help, as it was in Sweden till half a century ago. The article also explores how various aspects of solidarity-public and private-have developed and are changing in Sweden, known for its extensive welfare programs, with "from cradle to grave" security. It concludes that intergenerational solidarity has not vanished in Sweden; just the manifestations have changed. PMID- 17135093 TI - Danish home care policy and the family: implications for the United States. AB - This paper provides an overview of reforms in Danish long-term care initiated in the early 1980s, describes the relationship between elder care in Denmark and the family, and considers implications for U.S. policy. The success of Denmark's community-based experimentation with new models of home care and housing for the elderly resulted in a national decision to eliminate new construction of nursing homes and increase access to publicly funded home care. Lingering concern that the provision of paid assistance for the elderly could undermine family structure is allayed by the findings of a recent survey: Three-fourths of the elderly report seeing their children on a weekly or more frequent basis. Findings from the Danish experience provide evidence that community-based services can aid family caregivers, enable the frail elderly to live in the setting of their choice, and be cost-effective from a public policy perspective. PMID- 17135094 TI - A critical review of Singapore's policies aimed at supporting families caring for older members. AB - This article critically examines the family-oriented social policies of the Singapore government aimed at supporting families caring for older members. The sectors focused on are financial security, health, and housing. Singaporeans have been reminded that the family should be the first line of defense for aging families, followed by the community - the state would step in as the last resort. Drawing from recent research and examination of the state policies, the author argues that more should be done to help family caregivers looking after elder relatives. Recommendations for innovative ways to recognize and reward family carers conclude the paper. PMID- 17135095 TI - Families' values and attitudes regarding responsibility for the frail elderly: implications for aging policy. AB - This study examines the norms and values associated with care to disabled and frail aging parents, in particular those with regard to the sharing of responsibilities for care between families and formal services, and this within three age cohorts in Quebec, Canada. It is based on a telephone interview of 1,315 people. Factor analysis yielded four factors: (1) family responsibility; (2) uncompromising family obligations; (3) acceptance of services; (4) distrust of services. Analyses of the data indicate that all three age cohorts consider that families have responsibilities for their aging family members, at the same time that they score very high on the acceptance of service scale. This article discusses these seemingly paradoxical results and their implications for aging policy. PMID- 17135096 TI - Commentary: what role for the family and Medical Leave Act in long-term care policy? AB - The Family and Medical Leave Act provides unpaid leave but a key component is its intergenerational structure, permitting employees to take time off from work to care for an infant as well as an ill elderly parent. However, in an effort to provide paid leave, 23 of 28 states dropped the elder care provision in new initiatives introduced between 2000 and 2003. This article argues that in reforming leave policy in the future, the leave should be paid, remain intergenerational, cover more workers, and be adaptable to changing family patterns in an aging society. Also discussed is California's paid leave law, which meets these criteria. PMID- 17135097 TI - Preliminary experiences of the states in implementing the National Family Caregiver Support Program: a 50-state study. AB - Despite increased attention to policy choices to support family and informal caregivers, relatively little is known about states' experiences in providing caregiver support services. This article reports on the first nationwide survey of all 50 states and the District of Columbia in providing caregiver services since the passage of the National Family Caregiver Support Program. State program administrators reported that their program differs from other home and community based services because of the explicit focus on the family or informal caregiver. Results suggest that despite an increasing availability of caregiver supports in all 50 states, there is also a great unevenness in services and service options for family caregivers across the states and within states. PMID- 17135098 TI - Families, work, and an aging population: developing a formula that works for the workers. AB - This article examines the intersection of family caregiving, work, and long-term care. Supporting families who provide care in order to minimize negative work effects while enhancing the acceptability of care options is of common concern to employers, state and federal policymakers, and the homecare professionals in the community-based care system. The contribution of families to the long-term care system, how employer policies have developed, how the public policy agenda has addressed family caregiving, and the importance of a more effective partnership on the state level are discussed. PMID- 17135099 TI - Family and friends as respite providers. AB - Consumer-directed service options in home- and community- based care are increasingly available to adults with chronic conditions and cognitive impairments and to their family caregivers. Few studies, however, examine the experience of family caregivers who, when given a choice of providers of respite assistance (i.e., relief from the stress of providing constant care), prefer to hire family or friends rather than service providers. This study describes the in home respite experience of family caregivers served by California's Caregiver Resource Centers "direct-pay" program who hire family or friends (n = 39) or service providers (n = 77) to provide in-home respite assistance. Findings revealed similarities between the two groups with few exceptions: caregivers who hired family or friends reported poorer physical health, were slightly more satisfied with the respite assistance, and received more hours of respite at a lower unit cost. These findings lend support to consumer-directed respite service options where family caregivers are given flexible alternatives that may act to remove barriers to respite service availability and use. PMID- 17135100 TI - The family caregiving career: implications for community-based long-term care practice and policy. AB - Informal (i.e., unpaid) long-term care for disabled older adults is often chronic, but it is only recently that research has considered the longitudinal implications of family caregiving. In particular, investigators have conceptualized caregiving as a "career," and within the caregiving career, a number of diverse trajectories and transitions can occur. Following a summary of these findings, this paper considers how longitudinal caregiving research can influence and potentially address key policy and practice concerns, especially in the delivery and support of community-based long-term care (CBLTC) services. It is suggested that with the refinement of the informal long-term care literature, existing policy and practice to support caregiving families can be similarly advanced. PMID- 17135101 TI - Zoning, accessory dwelling units, and family caregiving: issues, trends, and recommendations. AB - This article explores the relationship between zoning regulations and co residential family caregiving in the United States. It first provides an overview of U.S. housing policies, especially zoning. We then describe major changes in family structure and composition in the United States with their implications for caregiving and discuss how multigenerational housing options, particularly accessory dwelling units (ADUs) in single-family homes, can help support family caregiving. After an overview of zoning policies and actions that inhibit ADU production, we document current trends, incorporating information from a small non-random study of ADU activity we conducted in 2004. Finally, we present recommendations for promoting more multigenerational housing as a supplement to other family support programs (e.g., dependent care assistance, family caregiver payments) and as a source of affordable, supportive housing for those families choosing co-residence as their eldercare solution. PMID- 17135102 TI - Resident and family perspectives on assisted living. AB - This research describes and compares the relative importance residents and family members place on attributes of the environment, the programs, and the policies of assisted living; describes their satisfaction with these features; and identifies factors associated with congruence between residents' and family members' ratings of importance and satisfaction. Both residents and their family members had high importance and satisfaction ratings. Family members gave the assisted living setting lower satisfaction ratings on all features than did residents. Congruence ranged from 34% to 71% for importance items and from 29% to 63% for satisfaction. Female residents, affectionate family relationships, and residing in an AL owned by a chain were positively associated with congruence on importance items, while resident and family education, resident income, and family involvement were negatively associated with congruence on importance items. For congruence on satisfaction items, having an affectionate relationship was positively associated and higher ADL dependency, more family involvement at the facility, and family members who viewed the facility as a safe place were negatively associated with congruence. This study makes a major stride forward because cognitively intact residents' perspectives are compared and contrasted with their own family members' perspectives, thus showing that residents and family members are two distinct groups, each with a unique set of preferences. PMID- 17135103 TI - Welfare reform: challenges for grandparents raising grandchildren. AB - This paper examines the impact of Temporary Aid to Needy Families (TANF) on a growing constituency that may require welfare assistance: grandparents raising grandchildren. A brief review of the demographics of grandparent caregivers is followed by an exploration of welfare reform legislation, including work requirements and the Healthy Marriage Initiative (HMI). The importance of state level policies in assisting grandparent-headed households, particularly health care, school enrollment, and housing policies, are also discussed. The paper concludes by offering implications for welfare policies in terms of their impact on intergenerational households headed by grandparents. PMID- 17135104 TI - State policy decisions in the 1990s with implications for the financial well being of later-life families. AB - This study explores trends and patterns in states' policy decisions affecting the economic well-being of later-life individuals and families in the United States in recent decades, focusing on the 1990s. Rules were selected from the areas of inheritance, estate taxes, homestead exemptions, Medicaid eligibility, estate recovery, and filial responsibility. Results indicate an increasing use of a broad definition of family, one implying that spouses, the nuclear family, extended kin, step-relations, and sometimes in-laws constitute an ongoing collective whose members share economic resources and risks over their lives and beyond. Despite this global trend, states varied in their rules addressing intrafamilial financial obligations and families' accountability to states. While some seemed interested in facilitating the conservation of familial resources, others seemed willing to minimize public assistance while coercing kin into accepting financial responsibility for one another. Research was suggested to answer questions raised by this study. PMID- 17135105 TI - Do determinants of medicare supplemental coverage choice vary by income. AB - Using data from the Community Tracking Study Household Survey (1998-99), we estimate the relationship between Medigap premiums and senior Medicare beneficiaries' supplemental coverage decisions. All seniors are more likely to be enrolled in an HMO in markets with higher Medigap prices. Lower income seniors are particularly sensitive to Medigap premiums and are more likely to have no supplemental coverage when faced with higher Medigap premiums. As Medicare supplemental options evolve in response to the 2003 Medicare Modernization Act, it is important to consider that lower income beneficiaries may respond to price changes and other factors differently than their higher income counterparts. PMID- 17135106 TI - The percentage of beds designated for medicaid in american nursing homes and nurse staffing ratios. AB - Previous analyses of the inverse relationship between a nursing home's Medicaid census and its quality of care have been based on samples limited to specific geographic regions, for-profit entities, or only skilled care facilities. The present study uses national-level data from the 1999 National Nursing Home Survey to examine the association between the proportion of beds designated for Medicaid residents and nurse staffing ratios. The results indicate that homes which designate a higher proportion of their beds for Medicaid recipients maintain lower ratios of registered nurses and nurse's aides to residents, even when key facility characteristics are controlled. It was also found that nursing homes with a higher proportion of Medicaid beds offer lower nursing ratios regardless of their profit status or the difference between private pay rates and Medicaid reimbursement rates. Since lower nursing ratios have been previously linked to negative outcomes, these findings suggest that homes which rely more heavily upon Medicaid recipients may be using cost-cutting strategies which have negative implications for quality. PMID- 17135107 TI - Discussion of quality and audit in health. AB - Attaining quality health care has long been a social policy priority for countries internationally. This discussion considers issues important to understanding quality, and audit implementation in particular. The paper covers, first, the principles and practice of audit and, second, broader implementation issues, which together point to the further development of quality initiatives in health in the United Kingdom health care context. To close, the future of audit as a means of improving health care is elaborated. PMID- 17135108 TI - Race, welfare reform and caseworkers' treatment of welfare recipients: an examination of an alternative service delivery system-the Wisconsin Works (W-2) Program. AB - This study examined how welfare recipients in the Wisconsin Works Program were treated by community-based organizations' caseworkers, as opposed to government agencies, to determine if racial disparities existed. The same racial disparities found with public agencies existed. And CBOs were not providing Blacks with services needed to get off welfare. PMID- 17135109 TI - Psychocultural influences on health care acceptability among elderly U.S. Pacific Islanders. AB - There is widespread recognition of the influence of ethnic variation on immigrant response to health care services, but far less is known about source of variation among nonimmigrant ethnic enclaves. Pacific Islander populations under U.S. administration for more than a century illustrate the potential influences of cultural factors on health care. Focus groups among elderly Samoan, Native Hawaiian, and Chamorro residents of southern California in 2002 found ethnic variation in such characteristics as expectations of publicly financed health care and in the willingness to discuss alternative sources of help with clinicians. These variations appear influenced by the colonial health care experience of these U.S. territories and, in the case of Samoan women, in distinctive perceptions of the role of prayer and traditional healing methods in care. Such psychocultural factors appear more potent than English language proficiency as an influence on the acceptability of health care among Pacific Islander elders. PMID- 17135110 TI - Childhood immunization coverage in US states: the impact of state policy interventions and programmatic support. AB - Although research suggests numerous interventions that can improve immunization coverage (Taskforce on Community Preventive Services, 2000), there is often a gap between policies supported by and public entities. The question for this study is whether the variation in childhood (19 to 35 months) immunization coverage rates across states is related to significant variations in state regulatory regimes that may optimize the benefits of state registries and systems that are designed to improve assessment of immunization practices. Utilizing 2002 data from the CDC and survey data collected from state immunization program officials, we find that financial support for state immunization programs, opt-out state registries, and state-mandated participation in provider quality improvement and assessment programs have positive associations with statewide coverage rates. We also suggest that more active state governmental support for interventions supported by rigorous scientific evaluation will not only improve early childhood immunization coverage, but may also support other public health objectives such as life-time full immunization and improve bioterrorism response planning. PMID- 17135111 TI - Birth defects as an indicator of the health status of Haitian women and their children. AB - This study focuses on the incidence of birth defects among children born to Haitian women in Florida. Analysis of comparative data collected by the Florida Birth Defects Registry indicates a disproportionate rate of birth defects among babies born to Haitian women. Sociocultural and economic characteristics of the Haitian community and their relationship to birth defects are explored. The implications of the findings for public policies that address prenatal care and childbirth- related problems are discussed. PMID- 17135112 TI - Sexual minorities and selection of a primary care physician in a midwestern U.S. city. AB - How and why sexual minorities select a primary care physician is critical to the development of methods for attracting these clients to a physician's practice. Data obtained from a sample of sexual minorities in a mid-size city in our nation's heartland would indicate that these patients are loyal when the primary care physician has a positive attitude toward their sexual orientation. The data also confirms that most sexual minorities select same sex physicians but not necessarily same sexual orientation physicians because of lack of knowledge of physicians' sexual orientation. Family practice physicians and other primary care physicians can reach out to this population by encouraging word of mouth advertising and by displaying literature on health issues for all sexual orientations in their offices. PMID- 17135113 TI - From "Male Bonding Rituals" to "Suicide Tuesday": A qualitative study of issues faced by gay male ecstasy (MDMA) users. AB - Use of MDMA and other club drugs has increased among gay men, but questions remain concerning the use, context, and popularity of these drugs; and views of their side effects. We interviewed in-depth 12 gay MDMA users in New York. MDMA had specific appeals to gay men, who often described isolation and stigmatization. Users underwent a period of initiation and social networks often became comprised of greater numbers of other users. Club environments fostered drug use and vice versa. Awareness of potential adverse effects varied and was minimized. Appropriate educational and prevention programs are needed to address these issues, and must take into account the specific contexts of these gay men's lives. PMID- 17135114 TI - Understanding the demographic characteristics of urban men who have sex with men. AB - We examined migration, closetedness, and family structure explanations of demographic profiles of urban men who have sex with men (MSM) using data from the Urban Men's Health Study and the Public Use Microdata Sample of the U.S. Census. The data suggested that the demographic profiles of urban MSM reflect a large in migrating white, often young adult, population without children. Closetedness, an oft-cited bias in MSM population studies, did not significantly impact demographic patterns with minor exceptions. MSM urban subpopulations have demographics that differ substantially from the general population of the cities in which they reside. PMID- 17135115 TI - Attempted suicide among transgender persons: The influence of gender-based discrimination and victimization. AB - To determine the independent predictors of attempted suicide among transgender persons we interviewed 392 male-to-female (MTF) and 123 female-to-male (FTM) individuals. Participants were recruited through targeted sampling, respondent driven sampling, and agency referrals in San Francisco. The prevalence of attempted suicide was 32% (95% CI = 28% to 36%). In multivariate logistic regression analysis younger age (<25 years), depression, a history of substance abuse treatment, a history of forced sex, gender-based discrimination, and gender based victimization were independently associated with attempted suicide. Suicide prevention interventions for transgender persons are urgently needed, particularly for young people. Medical, mental health, and social service providers should address depression, substance abuse, and forced sex in an attempt to reduce suicidal behaviors among transgender persons. In addition, increasing societal acceptance of the transgender community and decreasing gender based prejudice may help prevent suicide in this highly stigmatized population. PMID- 17135116 TI - Forced online: push factors of internet sexuality: A preliminary study of online paraphilic empowerment. AB - Online sexual behavior may be more completely explained by integrating a Pull Factor analysis, centering on the characteristics of the Internet that make it an appealing outlet for sexuality (e.g., "Triple A Engine") and a Push Factor analysis, centering on characteristics of the offline world that disallow fulfillment of sexual desires. This new framework is utilized in explaining paraphilic (unconventional) sexualities online. Because paraphilics are members of highly concealable, horizontal and rare sexual minorities, they are pushed online by mainstream society. Online, paraphilics interact with similar others, and through lurking, re-learning and self-disclosing, may become sexually empowered. Clinical ramifications of attainment of positive sexual-social identity via empowerment are discussed, as are possible affects on public policy. PMID- 17135117 TI - Layers of leather: The identity formation of leathermen as a process of transforming meanings of masculinity. AB - Leathermen form a gay male subculture that eroticizes leather dress and symbols. This investigation examined the relationship of participants' leather identity to their gender and sexual identities. In addition, the participants described their process of leather identity development, and its meanings and purposes. Six self identified leathermen participated in semi-structured interviews that were subjected to a grounded-theory analysis. The analysis suggested that leathermen develop a unique form of masculinity, integrating care and vulnerability with an aesthetic of heightened masculine appearance. Flexible interactional scripts allow for gendered signs to be enacted, designating a social status that is not recognized by the mainstream gay community. Findings are discussed in relation to previous research on gay male masculinity, gay community, sexual identity formation, and internalized homophobia. PMID- 17135118 TI - Infusing lesbigay research into the counseling research classroom. AB - Despite research findings that lesbigays value, want, and use counseling services, some counselors report they are not adequately prepared to work with the lesbigay population. Lesbigay students have reported biases and discrimination in their programs. Research has found that some mental health professionals engage in a number of harmful practices when working with lesbigay clients. These findings challenge counselor education programs to find better ways of infusing lesbigay issues into their curricula. This article discusses how lesbigay issues might be infused into the counseling research classroom in ways that provide students the opportunity to think critically; learn about research design and ethics; expand their levels of tolerance; develop a deeper appreciation of the work involved in doing research; and see the power of research to change the course of history. PMID- 17135119 TI - College students' multiple stereotypes of lesbians: A cognitive perspective. AB - This paper examines stereotypes of lesbians held by college students. Multiple stereotypes are elicited from a free response trait listing task, followed by a sorting task. The results of the sorting task are submitted to cluster analysis and multidimensional scaling to reveal the complexity of cognitive representations of this group. Eight types are described, reflecting underlying distinctions between positive perceptions (e.g., lipstick lesbian, career oriented feminist) and negative perceptions (e.g., sexually deviant, angry butch) and also between relative strength and weakness. The research is discussed in terms of cognitive perspectives on stereotyping and gender inversion theory. Suggestions for future research are provided. PMID- 17135120 TI - Reactive, anxious and possessive forms of jealousy and their relation to relationship quality among heterosexuals and homosexuals. AB - The present study examined the relationship between relational quality and three different types of jealousy-reactive, anxious and possessive jealousy. The sample consisted of 76 gay men, 79 lesbians, 70 heterosexual women and 70 heterosexual men. Findings show that different types of jealousy affect relationship quality differently and do so differently for homosexuals and heterosexuals. Among heterosexuals and especially gay men--but not among lesbians--anxious jealousy was negatively related to relationship quality. In contrast, among heterosexuals- but not among gay men or lesbians--reactive jealousy was positively related to relationship quality. The present study shows that distinguishing between different types of jealousy is necessary to entangle the diverse effects of jealousy on the quality of homosexual and heterosexual relationships. PMID- 17135121 TI - Commitment in same-sex relationships: A qualitative analysis of couples' conversations. AB - Theoretical constructs and meanings of relational commitment for same-sex couples have typically been generalized from heterosexual relationships. Same-sex couples, however, face a unique set of challenges in constructing committed relationships. To expand our knowledge of the meaning of commitment, same-sex couples described their lived experiences in defining and creating a committed relationship. Transcripts of the conversations of 14 same-sex couples (7 male and 7 female couples) were subjected to analysis using the Consensual Qualitative Research (CQR) method (Hill, Thompson, and Williams, 1997). Seven domains emerged, revealing that these same-sex couples constructed the meaning of commitment through comparisons, costs, intra-couple differences, investments, personal and relationship values and ideals, rewards, and sexual boundaries. Unique aspects of commitment are discussed as well as the implications of these findings for future research and service delivery. PMID- 17135122 TI - Translators, traitors, and traducers: Perjuring Hawaiian same-sex texts through deliberate mistranslation. AB - In the long history of the West's encounter with Hawaiian culture, which began in the late 1700s with Captain Cook, translators and translations have often been the tools of intentional falsehood, thus demonstrating the truth of the Italian proverb, Traduttore, traditore ("the translator is a traitor")--particularly with regard to same-sex texts. The standards of truth have often been subverted in translation by the demands of foreign religion, hegemony, business, and academe. This subversion continues to this day in the form of the "missionary mentality" in politics and law. The way out of this situation is a brutally honest cleaning off of the besmirched Hawaiian texts. PMID- 17135125 TI - Gay priests and other bogeymen. AB - The sexual abuse of boys by priests was at the center of the 2002 scandal in the Roman Catholic Church. This scandal has resurrected the misconception that a link exists between having a homosexual orientation and being at increased risk for being a pedophile or child molester. This paper reviews and debunks the arguments in support of this misconception. Central to these arguments is what might be called the "proportionality argument": that the ratio of homosexuals to heterosexuals among child molesters is higher than the ratio of homosexuals to heterosexuals in the general population. The flaws in the proportionality argument and several other misconceptions are discussed. PMID- 17135126 TI - Can one TV show make a difference? Will & Grace and the Parasocial Contact Hypothesis. AB - Television has an opportunity to influence beliefs about groups with which individuals typically may have little direct social contact. This study describes a synthesis of the Contact Hypothesis and the concept of Parasocial Interaction to pose what we call the Parasocial Contact Hypothesis to test whether exposure to gay men on Will & Grace can influence attitudes toward gay men in general. Based on a study of 245 university students, this study examines the relationships among number and intimacy of gay social contacts, parasocial interaction, viewing frequency of Will & Grace, and scores on Herek's Attitudes Toward Gay Men and Lesbians scale. Increased viewing frequency and parasocial interaction were found to correlate with lower levels of sexual prejudice-a relationship that was most pronounced for those with the least amount of social contact with lesbians and gay men. PMID- 17135127 TI - Sexuality and school shootings: what role does teasing play in school massacres? AB - Conventional explanations - lax gun control laws, media violence, single and working parents - do not adequately explain the recent spate of school shootings, and neither does bullying by itself, an explanation recently gaining more traction. A certain type of bullying, however, is revealed as particularly culpable. Many of the recent shootings share a disturbing component: The perpetrators were repeatedly, even relentlessly, accused by "preps and jocks" of being gay. Gay harassment is proposed as a point of departure for understanding the causes of school shootings. When boys who believe on some level that they warrant privilege, are instead harassed, they may feel driven to avenge the "wrong," and re-assert a more dominant, powerful, and victorious masculinity. These circumstances call for a cultural transformation such that "boys will be boys" is no longer used as an alibi for violence. The author presents implications for school-based social work practice, teacher and administrator interventions, as well as other prevention strategies. PMID- 17135128 TI - Attitudes, perceptions, and knowledge of pre-service teachers regarding the educational isolation of sexual minority youth. AB - Sexual minority youth (Gay, Lesbian, Bisexual, Transgendered, and Questioning youth) must function in a society that condones homophobia. Rendered invisible through the stigma attached to their identification as sexual minorities, they are conveniently forgotten by the institutions charged with facilitating their education. Results from a research study conducted by Sears (1991), indicated that teachers often expressed that they should be more proactive and supportive and committed to the welfare of all of their students. However, the realities of their professional intervention and support were negligible. The purpose of this research study was to examine the personal beliefs of pre-service educators from the Midwest about homosexuality, and how these attitudes and feelings are actualized in the schools. Findings suggest that the results of the current sample had little variation from the sample reported by Sears. While Black pre service educators as well as early childhood pre-service educators obtained higher mean scores on both attitudinal measures (Attitudes Toward Homosexuality and Index of Homophobia), race was the only variable which proved to be statistically significant. Implications for future research are also discussed as a result of this study. PMID- 17135129 TI - A voice in the wilderness: gay and lesbian religious groups in the Western United States. AB - The paper describes a study of gay and lesbian religious groups located in the United States between the Mississippi River and the Pacific Ocean between March of 1989 and June of 1993. The researcher found 684 groups to have existed at some time in the region and conducted interviews to document 468 of these groups. While a history of these groups dates back to the 1930s in the region, large groups still existing today did not begin to form until the late 1960s. The six most common goals of the groups were social and support needs, worship, denominational and social politics, community service, and leadership in the gay and lesbian community. Being focused on gay and lesbian issues has an influence on when and how they came into existence (in the beginnings of the gay and lesbian movement, in response to the AIDS crisis, in response to anti-gay legislative initiatives), has an influence on the kinds of key themes addressed (ordination of gay and lesbian people, same-sex marriage, service for people with AIDS, sexual ethics in scripture), and has an influence on relationships with the larger community (shunning or inclusion in the larger society and in the larger religious community). However, the stronger influence is their identity as religious (and, therefore, social) institutions. They experience the same pressures of mainstream religion and as social institutions by facing issues of diversity, gender, age, politics, race, and disease. They face the same challenges as any religious institution today of being prophetic, responsive, changing, traditional, true, relevant, and even noticed. A distinct feature is their role on the fringes. They are on the fringe of religious life and on the fringe of gay and lesbian life. PMID- 17135130 TI - Why is their picture on the wedding page?: A rural community responds to a union announcement. AB - On August 17, 2003, two women placed a union announcement in the Grand Forks Herald and became the first same-sex couple in North Dakota to do so (Lee, 2003). This announcement generated a flood of letters, both pro and con, that appeared in on the editorial page in the Herald over the next several weeks. In this exploratory study, I conduct a content analysis of those letters. Through this analysis, the three primary thematic categories emerged upon which writers based their arguments. They include those grounded in issues of tolerance, arguments based on Christian values and Scripture and those that focused on normalization. Additionally, the letters reveal something about rural versus urban sensibilities in that relatively more urban Grand Forks residents were less likely to write negative letters than their counterparts from neighboring small towns. I conclude with a discussion of the implications of these findings and suggestions for future research. PMID- 17135131 TI - Same-sex domestic partnerships and lower-risk behaviors for STDs, including HIV infection. AB - To determine the association between domestic partnership status and risk behaviors for sexually transmitted diseases, including HIV infection, we analyzed data from a population-based interviewer-administered telephone survey of 2,881 gay men in San Francisco, Los Angeles, New York and Chicago conducted in November 1996 to February 1998. Men in domestic partnerships had a statistically significantly lower prevalence of multiple partnerships, "one-night stands," and unprotected anal intercourse with a non-primary partner than either men with steady partners not identified as domestic partners or men without a steady partner. These findings were independent of age. Men in domestic partnerships had decreased risk behaviors for sexually transmitted diseases, including HIV infection, suggesting but not proving, that conferring legal status to same-sex unions might decrease sexual risk behavior. PMID- 17135132 TI - Psychology graduate students' attitudes toward lesbians and gay men. AB - The extant literature concerning attitudes toward gay men and lesbians was reviewed, with a particular emphasis on the attitudes of mental health professionals. Seventy graduate students in a variety of psychology-related areas were surveyed about their attitudes toward lesbians and gay men. Findings from hierarchical multiple regression analyses indicated that after controlling for socially desirable responding, the only variable to predict negative attitudes toward gay men and lesbians was endorsement of traditional gender roles. The results of this investigation were placed within a theoretical framework and then implications for training future mental health practitioners were discussed. PMID- 17135133 TI - Fraternal birth order and ratio of heterosexual/homosexual feelings in women and men. AB - Studies of the 2-3% of persons who identify as homosexual found men but not women had more older brothers than persons who identify as heterosexual. The present study investigated the birth order in the approximately 20% of men and women who anonymously report some homosexual feelings, few of whom identify as homosexual. The number of older brothers and sisters was investigated in seven cohorts: 319 male twins; and 49, 54, and 61 female and 66, 116, and 50 male medical students. Both women and men who anonymously reported homosexual feelings had a greater mean number of older brothers and sisters than did those who reported no homosexual feelings. The difference was stronger in relation to brothers than sisters. The birth order effect was not related to the strength of the subjects' degree of homosexual compared with heterosexual feelings. Its presence in women could not be accounted for by the widely accepted hypothesis that the birth order effect is due to a maternal immune reaction provoked only by male fetuses. The lack of relationship between the strength of the effect and degree of homosexual feelings in the men and women suggests the influence of birth order on homosexual feelings was not due to a biological, but a social process in the subjects studied. Investigating the neglected significant percentage of predominantly heterosexual men and women who anonymously report some homosexual feelings may aid in understanding the factors influencing sexual orientation, and identity. PMID- 17135135 TI - Offering answers and insight into occupational health dangers for a variety of populations. PMID- 17135136 TI - Incidence of hematologic malignancies in agriculture. PMID- 17135137 TI - Pesticides: what is the role of toxicovigilance. PMID- 17135138 TI - Barriers and benefits of protective eyewear use by Latino farm workers. AB - Agricultural work is one of the riskiest occupations for the eyes. Protective eyewear can prevent eye injuries in upward of 90% of cases. Latino migrant and seasonal farm workers (LFWs) are at particular risk of eye injury because of economic, social, and cultural barriers to safe employment. The goal of this project was to determine the perceived benefits and barriers to use of protective eyewear during agricultural work among LFWs. In the year 2000, 55 workers who spent at least part of their time working in Illinois and Michigan as seasonal, hired farm workers were interviewed either individually (9) or in six focus groups (46) regarding their perceptions of eye hazards and the factors that encourage or discourage the use of protective eyewear. Subsequently, safety glasses and training on eye safety were delivered by promotores de salud to LFWs from the same cohort over two summer seasons. During the second season, the promotores observed the use of safety eyewear by farm workers at least once per week on 17 farms over a three-month period. At each point of observation, the promotores asked those wearing the glasses why they chose to wear them and those who were not wearing them, why not. At both points of the study (pre-intervention interviews and post-intervention observations), the reasons for use fell into the following categories: protection from hazards, appearance, whether it was mandated/provided by the employer, and whether others used it. The reasons for non-use include perceived lack of protection, discomfort, undesirable appearance, interference with visual acuity, slowing down the work pace, and no mandate from employers. Since eliminating the eye injury hazard is not possible in most agricultural settings, administrative strategies and use of personal protective equipment are critical. Perceptions of the target audience, LFWs, should be used to develop a comprehensive eye safety program; this should include allowing LFWs to select a style from effective prototypes, providing eyewear and promoting/mandating its use in hazardous job tasks, and implementing a comprehensive PPE program for eye safety. In addition, addressing functional problems - falling off, fogging, loss and forgetting glasses, the pace slowdown that reduces production and leads to lower wages for workers - should be addressed. PMID- 17135139 TI - Training methods and association with worker injury on Colorado dairies: a survey. AB - The goals of this study were to define the current methods for training entry level workers on Colorado dairies, to measure the relationship of training methods to the incidence of work-related injuries on those operations, and to characterize the nature and cause of injuries reported by participating workers. Seventy-two workers from 15 dairies from Weld, Morgan, Larimer and Yuma counties volunteered to be interviewed privately in the language of their choice. Training methods were classified by content (task training versus safety training), method of delivery (on-the-job verbal instruction, on-the-job demonstration, on-the-job supervision, classroom- based, and / or reading materials); and instructor (supervisor and/or co-worker). The majority (84.7%) of interviewed workers were Mexican-born, Spanish-speaking males. Most workers reported the training to be a combination of verbal instruction, demonstration and supervision. Twenty-nine of 72 (40.3%) of all participants suffered at least one work-related injury in the 12 months prior to the interview. Entry-level safety training had no protective effect (P = 0.463) on the reported incidence of injury in dairy workers in the previous year. However, workers that received task-related training were four times less likely to suffer a work-related injury (P = 0.008). Training conducted by a co-worker also had a significant protective effect against work-related injuries (P = 0.028). Dairy workers identified direct contact with livestock or machinery as the main cause for work-related injuries, and most injuries occurred while medicating sick cows and milking. PMID- 17135140 TI - Treating skin disease: self-management behaviors of Latino farmworkers. AB - Latino migrant and seasonal farmworkers experience high rates of skin disease that result from their working and living conditions. Knowledge of the ways farmworkers treat skin disease symptoms will provide a foundation for developing culturally appropriate health education, improving the delivery of health services, and improving occupational health policy for agricultural workers. The purpose of this paper is to describe skin disease self-management practices among Latino migrant and seasonal farmworkers in North Carolina. This analysis uses a qualitative design based on in-depth interviews with 30 Latino farmworkers (six females, 24 males). Computer assisted, systematic procedures are used to analyze the verbatim transcripts of these interviews. Participants shared a consistent set of health self-management actions in treating skin disease. These actions were within the domains of self-care and medical care. A model of skin disease self-management among Latino farmworkers includes the self-care actions of hygiene, use of home remedies and use of over-the-counter remedies, with farmworkers often combining different domains of self-care. While farmworkers acknowledge the benefits of medical care, they are also mindful of barriers to its use, including cost, transportation and language. The large percentage of farmworkers who experience skin problems indicates that health outreach workers who serve this population need to provide education on preventing and treating skin problems, and they need to recommend to farmworkers appropriate over-the counter medicines for the treatment of these skin problems. Appropriate medical care for treating skin problems that are dangerous and reduce farmworkers' quality-of-life needs to be made available to this population. PMID- 17135141 TI - Spanish-speaking dairy workers in New York, Pennsylvania, and Vermont: results from a survey of farm owners. AB - A New York Center for Agricultural Medicine and Health (NYCAMH) study surveyed 294 dairy farms in New York, Pennsylvania, and Vermont. The study utilized a quarterly telephone survey to assess the proportion of Spanish-speaking workers on these farms, and also to contrast the hazard level of work tasks and prevalence of lost work time between Spanish- and English-speaking workers. The total workforce followed in the study was comprised of 14.4 percent Spanish speaking workers, with larger farms having a higher proportion than smaller farms (19.9% versus 4.6%, respectively). Of the 294 farms, 22.5 percent had at least one Spanish-speaking worker, which differed, greatly between larger and smaller farms (51.5% versus 7.3%). Spanish workers were significantly younger, worked significantly longer hours and had significantly fewer years of employment than their English-speaking counterparts. Work hour differences were more pronounced on the larger farms. Lost work time, due to on-farm injuries, did not differ between the Hispanic workers and the non-Hispanic workers. After correcting for both age and length of farm employment, Spanish-speaking workers were far less likely to perform managerial functions than their English-speaking counterparts (OR = . 22 p < .01). PMID- 17135142 TI - Ache, pain, and discomfort: the reward for working with many cows and sows? AB - The main purpose of the study was to investigate the prevalence of perceived symptoms of musculoskeletal disorders (MSD) among workers on large-scale dairy and pig farms in Sweden (herd size more than 300 cows and 450 sows) and to identify potential risk factors in the development of MSD. A study based on questionnaires was carried out among 42 workers on 10 large dairy farms and among 37 workers on 10 large pig farms in Southern Sweden during the autumn of 2002. Most importantly, the study showed that 86% of the dairy workers and 78% of the pig workers reported some kind of MSD during a period of 12 months prior to the study. The most frequently reported MSD among both the dairy and the pig workers were in the "upper extremities" (52% and 62%, respectively) especially in the shoulders and in "the back" (60% and 57%, respectively) especially in the lower back. Furthermore, being of short stature, doing repetitive work, working in awkward positions and being exposed to dust were significant risk factors in having MSD among the workers in this study. Thus, working with many cows and sows on large-scale farms in Sweden can be considered as a high risk job with regard to MSD. PMID- 17135143 TI - Fatal work-related injuries in the agriculture production sector among youth in the United States, 1992-2002. AB - Youth working on farms face unique risks that are not present for many other young workers, including machinery, large animals, electrical hazards, chemical hazards and excessive noise. This research identified the number and rate of occupational fatalities for youth working in the agriculture production industry, which is most closely affiliated with farming, for the years 1992-2002. The Census of Fatal Occupational Injuries (CFOI), developed by the Bureau of Labor Statistics (BLS), was the database used for the analysis. There were 310 work related deaths to youth less than 20 years of age from 1992 through 2002 in the agriculture production sector. This compares to 1,958 total fatalities for all workers less than 20 years of age for the same time period. The number of agricultural production fatalities to youth has shown a general downward trend over this time period. The rates were higher for young workers in agriculture production than for young workers in all industries by a factor of 3.6. Fifteen year olds had the highest fatality rates with the crop production sector having a rate six times that of all 15 year old workers. The objective of this descriptive research was to identify, prioritize and publicize the risks to children and youth who work on farms in order to provide public health and safety professionals relevant information upon which to base decisions for interventions or other prevention activities for this priority population. This research also has direct applications for farm parents and safety and health professionals who work with the priority population of young agricultural workers. PMID- 17135144 TI - California surveillance for pesticide-related illness and injury: coverage, bias, and limitations. AB - The California Pesticide Illness Surveillance Program (PISP) is a major resource for pesticide illness epidemiology. This work attempts to improve characterization of pesticide illness in California, evaluate case ascertainment of the PISP and identify PISP's limitations and biases for studying the incidence and epidemiology of pesticide-related illness. Abstractors collected standardized information from 1994-1996 PISP files, poison control logs, hospital records, and death certificates. Linkage produced a merged file of all records that documented consideration of pesticide exposure as a cause of ill health. We identified 23 deaths and estimated 1,310 hospitalizations attributable to pesticide exposure, corresponding to 0.024 fatalities and 1.38 hospitalizations (95% confidence interval [CI] = 1.01-1.74) per 100,000 California population person-years. Most intoxications followed domestic exposures and were not reported to the surveillance program, which identified primarily occupational exposures. Excluding groups of five or more, we estimated 16% ascertainment (CI = 0.05-0.27) for non-agricultural occupational cases. Agricultural connections increased the probability of ascertainment to about 50%, cultural and economic barriers notwithstanding. Surveillance records existed for all identified episodes in which five or more people were exposed. California pesticide surveillance data appear complete for mass exposures, and adequate to characterize agricultural and occupational exposures. Work is needed to explore domestic exposures and occurrences beyond the reach of the health care system. PMID- 17135145 TI - Effectiveness of cleaning practices in removing pesticides from home environments. AB - The deposition of agricultural pesticides in the homes of agricultural workers and residents of agricultural communities is a major environmental health concern. The effectiveness of home cleaning activities in removing pesticides from home surfaces has not been tested. An intervention study was conducted to assess the effectiveness of cleaning windowsills, floors and carpets in a sample of 10 farmworker homes. Baseline measures of organophosphorus (OP) pesticide residues were obtained, a standardized cleaning intervention was applied and follow-up measures of pesticide residues were obtained within 24-48 hours after the cleaning and 12 months later. House dust was analyzed for six OP pesticides. All homes had detectable baseline levels of OP pesticides on floors and windowsills. Cleaning of linoleum floors was ineffective in removing total pesticide residues and cleaning effectiveness varied among the pesticides. The cleaning of total OP pesticides on the windowsills was effective (median decrease was 0.0029 microg/cm(2), 1-sided p-value = 0.01). Steam cleaning carpets essentially reduced the amounts to non-detectable levels. In 12 months the levels in carpets had accumulated to one-third of the baseline levels. These results provide evidence that cleaning practices can reduce the amount of pesticides in agricultural homes; however the type of surface being cleaned and the pesticides present in the home may influence results. PMID- 17135147 TI - Using a portable wireless computer lab to provide outreach training to public health workers. AB - Librarians at Louisiana State University Health Sciences Center in Shreveport developed an outreach program for public health workers in north Louisiana. This program provided hands-on training on how to find health information resources on the Web. Several challenges arose during this project. Public health units in the region lacked suitable teaching labs and faced limited travel budgets and tight staffing requirements, which made it impractical for public health workers to travel. One solution to these problems is a portable wireless computer lab that can be set up at each site. The outreach program utilized this approach to present on-site training to public health workers in the region. The paper discusses operational and technical issues encountered in implementing this public health outreach project. PMID- 17135148 TI - Information literacy and library attitudes of occupational therapy students. AB - Information literacy, often described as a person's ability to effectively find and evaluate answers to questions using a variety of information resources, is of particular importance to health care workers. This paper presents the results of an information literacy survey presented to occupational therapy (OT) students at Thomas Jefferson University during a series of required class activities. Also described are the authors' activities with the faculty and courses at Jefferson. The survey was made available to first-, second-, third-, and fourth-year occupational therapy students along with nursing students and pharmacy students. The survey is designed to identify research habits, skills, and preferences. Results confirm some commonly held perceptions about searching skills of young adults and an interesting dichotomy in students' learning habits. The paper concludes with a discussion of recommendations to OT faculty and librarians on how to improve information literacy education. The survey can be obtained by contacting the authors. PMID- 17135149 TI - Evaluation of a hospital medical library class for NICU nurses. AB - A library class was designed and offered to new nurses from the Neonatal Intensive Care Unit at the Children's Hospital of Michigan between 2003 and 2005. The class was intended to increase their knowledge of quality health information resources and to assist them with their smooth transition to a new health care organization. The goal of the library training class was to develop the nurses' awareness and knowledge of the library services and online resources on the organization Intranet and to improve their skills in finding reliable information related to patient care, patient parent education, and research. An evaluation study was conducted to assess the effectiveness of the library class. Although the findings demonstrated strengths of the library class, they also revealed some areas for improvement. The data gathered resulted in a number of recommendations regarding library instruction design and evaluation. PMID- 17135150 TI - CIRRIE: an international rehabilitation resource. AB - The CIRRIE Database of International Rehabilitation Research is an appropriate companion resource to NARIC's REHABDATA Literature Database. It is produced at the University of Buffalo, State University of New York (or SUNY-Buffalo to those from the Northeast). CIRRIE includes only published information resources that cover research that is conducted outside of the United States. Those who wish to locate information about published research conducted within the United States should consult REHABDATA. This column presents basic information about the database, its searching conventions and thesaurus, and some general information about CIRRIE as a general resource of online information. PMID- 17135151 TI - Getting the picture: medical images on the Web. AB - Visual representations are used to convey ideas and concepts. They add value to educational venues. Files digitized from hard-copy media or born-digital files are standard formats, and the Internet is a resource for locating billions of these files. Searchers should be cognizant of the legal and ethical implications of using images downloaded from the Internet. This article intends to inform users about medical images on the Internet, as well as providing a select list of Web-based medical image sources. PMID- 17135152 TI - Librarians collaborating with nurses. AB - Nurses have the potential to be one of the largest groups of users a medical library can serve. In recognition of this fact, librarians at St. John's Health System embarked on a journey to collaborate with nurses and increase this group's use of the medical library. In 2004, a nursing outreach plan was developed by librarians, with input from non-physician health care employees. The nursing outreach plan will be reviewed along with the barriers and successes encountered during implementation of the plan. Innovative strategies for reaching nursing personnel were accomplished by partnering with nurses to reach nurses. PMID- 17135153 TI - e-Learning Object Portals: a new resource that offers new opportunities for librarians. AB - e-Learning Object Portals are emerging as a new form of electronic information. Four portals most effective in providing health care information are identified: The Medical Library Association's Center of Research and Education (CORE), the Health Education Assets Library (HEAL), the American Academy of Medical College's MedEDPortal, and the Multimedia Education Resource for Learning and Online Teaching (MERLOT). Various user groups such as librarians, educators, and students are discussed, as well as their reasons for using e-learning object portals. New roles for librarians in promoting and developing these portals are also reviewed. PMID- 17135156 TI - Antioxidant activity of Pepticare, a herbomineral formulation, in experimentally induced renal and cardiac damage. AB - Pepticare, a herbomineral formulation, was administered orally to rats at the dose levels of 125, 250, 500 and 1000 mg/kg to investigate its effect on isoproterenol-induced myocardial infarction and cisplatin-induced renal damage. The drug reduced the levels of serum creatine kinase (CK), glutamic oxaloacetate transaminase (GOT), lactate dehydrogenase (LDH) and uric acid in isoproterenol induced cardiac damage. In cisplatin-induced renal damage, Pepticare reduced the serum levels of creatinine, urea, blood urea nitrogen (BUN) and uric acid. It was further found that administration of Pepticare increased the levels of superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), membrane bound enzymes like Ca2+ ATPase, Mg2+ ATPase and Na+ K+ ATPase and decreased lipid peroxidation (MDA) in heart and kidney, respectively. Thus it can be concluded that Pepticare possesses antioxidant activity and protects the heart and kidney from damage caused by isoproterenol and cisplatin, respectively. PMID- 17135157 TI - Antisecretory and antiulcer activity of Asparagus racemosus Willd. against indomethacin plus phyloric ligation-induced gastric ulcer in rats. AB - OBJECTIVE: To study the antisecretory and antiulcer activity of Asparagus racemosus Willd. (methanolic extract) and its action against indomethacin (a non steroidal anti-inflammatory drug) plus pyloric ligation (PL)-induced gastric ulcers in rats. METHOD: Indomethacin plus PL-induced gastric ulceration model was used in the study. RESULTS: Treatment with Asparagus racemosus (Shatavari) crude extract (100 mg/kg/day orally) for fifteen days significantly reduced ulcer index when compared with control group. The reduction in gastric lesions was comparable to a standard antiulcer drug Ranitidine (30 mg/kg/ day orally). Crude extract also significantly reduced volume of gastric secretion, free acidity and total acidity. A significant increase in total carbohydrate (TC) and TC/total protein (TP) ratio of gastric juice was also observed. No significant change in the total protein was noted. CONCLUSION: Asparagus racemosus was found to be an effective antiulcerogenic agent, whose activity can well be compared with that of ranitidine hydrochloride. The results of this study suggest that Asparagus racemosus causes an inhibitory effect on release of gastric hydrochloric acid and protects gastric mucosal damage. PMID- 17135158 TI - Clinical evaluation to assess the safety and efficacy of coded herbal medicine "Dysmo-off" versus allopathic medicine "Diclofenac sodium" for the treatment of primary dysmenorrhea. AB - The purpose of the present research work was to carry out clinical study on primary dysmenorrhea to comparatively examine the coded herbal drug formulation "Dysmo-off" with authentic allopathic medicine "Diclofenac sodium" (NSAIDs). A random controlled clinical trial was conducted to compare the efficacy and safety of coded herbal medicinal treatments Dysmo-off with Diclofenac sodium/Phenylacetic acid. These evaluations were based on verbal rating scale so as to ascertain the rate of analgesic effects on dysmenorrhoeic pain. The patients were randomly allocated with the ratio of 1:2 for controlled treatment with nonsteroidal anti-inflammatory drugs (NSAIDs) (n = 40) received Diclofenac sodium tablets twice daily for 4 days (50 mg one day prior to and three days after the menstruation), and test treatment with Dysmo-off (n = 80) received powdered Dysmo-off twice daily for four days (5 g one day prior to and three days after the menstruation). Treatment lasted for 4 consecutive menstrual cycles. Hemoglobin, ESR and ultrasound were measured at baseline during study. All subjects were clinically studied and completed the assigned therapy during the period May 2001 to June 2004. PMID- 17135159 TI - Fatty acid composition of some medicinally useful seeds. AB - Fatty acid composition of seeds from the fruits of Butea monosperma, Jatropha glandulifera and Portulaca oleracea of three different families, namely Papilionaceae, Euphorbiaceae and Portulaceae were determined through Gas Liquid Chromatography to find novel natural sources of essential fatty acids for human health and of economic interest. Comparative studies were also performed to ascertain the utilization of each species for domestic as well as industrial purpose and the quality parameters developed can be utilized as marker characters for the aforesaid seeds used by the pharmaceutical industries. PMID- 17135160 TI - Protective effect of Indigofera aspalathoides against CCl4-induced hepatic damage in rats. AB - The alcoholic extract of stem of Indigofera aspalathoides was evaluated for its antihepatotoxic activity against CCl(4)-induced hepatic damage in rats. The activity was evaluated by using biochemical parameters, such as serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), total bilirubin and gama glutamate transpeptidase (GGTP). The histopathological changes of liver sample were compared with respective control. The extract showed remarkable hepatoprotective effect. PMID- 17135161 TI - Tyrosinase-inhibitory activity in some species of the lichen family Graphidaceae. AB - Twenty-five species of the lichen family Graphidaceae have been investigated for tyrosinase-inhibitory activity. Tyrosinase-inhibiting material was extracted with solvents methanol, acetone, ethanol, dimethyl sulphoxide in water, and with water only. Methanol has been found to be suitable for extracting adequate amounts of tyrosinase-inhibiting component from the natural thallus. The lichen species such as Graphina glaucorufa, Graphina multistriata, Graphina salacinilabiata, Graphis assamensis, Graphis nakanishiana, and Phaeographopsis indica, have shown inhibition of tyrosinase over a range of 30-78%. Half-inhibiting concentration (IC(50), microg/ml) has been found to be much lower than the standard tyrosinase inhibitors and thus can compete with other commercially available tyrosinase inhibitors. The extracts of these species have been found to be stable at 4 degrees C. PMID- 17135163 TI - Frequently asked questions (FAQ). AB - As Americans use botanical medicines in ever-increasing numbers, they are moving beyond capsules and tinctures, and turning to assorted preparations that will complete their herbal medicine chest. According to The Arthritis Foundation, almost 45% of patients apply ointments or rubs for osteoarthritis. Topical preparations have a long history in traditional herbal medicine systems, and make up a significant proportion of the armamentarium of indigenous practitioners. Although contemporary patients are often resistant to the inconvenience of topical preparations, they often will become compliant after a notable success. The nomenclature of these topical preparations, as one would expect, is murky. There are no clear and consistent definitions for the assorted terms, and different sources will use different terms for the same material or process. PMID- 17135164 TI - Devil's Claw (Harpagophytum procumbens DC): an evidence-based systematic review by the Natural Standard Research Collaboration. AB - An evidence-based systematic review including written and statistical analysis of scientific literature, expert opinion, folkloric precedent, history, pharmacology, kinetics/dynamics, interaction, adverse effects, toxicology, and dosing. PMID- 17135165 TI - The elusive romance of motherhood:drugs, gender, and reproduction in inner-city distressed households. AB - This paper explores the social contexts of reproductive decision making among poor African-American women in inner-city distressed households by focusing on women's narratives of their reproductive and maternal experiences. We explore the hidden agendas and motivations that underpin women's reproductive decisions and perceived choices within the turmoil of poverty, domestic instability, economic uncertainty, and addiction. The political economy of reproduction, within which birthing and motherhood in distressed inner-city households take place, generates the conditions for absent fathers, brittle unions, and a highly skewed gendered division of parenting. Locally constituted notions of gender, agency and autonomy are key dimensions in the cultural constructions of motherhood in these female headed households. Woven into the local maternal experiences is also the desire to 'give and receive love'. By focusing on women's own formulations of responsibility and agency in their reproductive decisions, we can see how they make sense of their reproductive histories and maternal experiences amidst the constraints of poverty, class, race, and substance abuse. PMID- 17135166 TI - Religiosity, refusal efficacy, and substance use among African-American adolescents and young adults. AB - Research points toward multiple pathways (i.e., psychosocial domains) through which religiosity influences substance use behaviors. This study examined whether refusal efficacy mediated the relationship between religiosity and substance use in African-American adolescents and young adults. Four hundred thirty-five urban and rural African-Americans, aged between 12 and 25, completed measures of private and public religiosity, refusal efficacy, and substance use (i.e., tobacco, alcohol, marijuana, and other illicit drug use). Tests for mediation were computed with private and public religiosity as independent variables, drug refusal efficacy as mediator, and substance use as the criterion. Results show that drug refusal efficacy mediated the relationship between private religiosity and tobacco, marijuana, and other illicit drug use, but not for alcohol use. Refusal efficacy also mediated the relationship between public religiosity and alcohol use, but not for other licit and illicit substances. The findings provide support for the unique impact of public religiosity when considering its role in preventive intervention. Future research should consider examining other psychosocial domains which may mediate the effect of religiosity on substance use behaviors among African-American adolescents. PMID- 17135167 TI - Sisters in support together against substances (SISTAS): an alcohol abuse prevention group for Black women. AB - This paper examines the existing literature for alcohol abuse prevention and treatment of African American women and offers a group counseling intervention to address the culturally specific needs of this population. The group, SISTAS: Sisters In Support Together Against Substances, includes a curriculum that focuses on key components such as spirituality, self-esteem, racial identity, social support and intimate relationships. The purpose of the group is to raise awareness of and educate members about the factors that can lead African American women from drug abuse to dependency. PMID- 17135168 TI - Unraveling the layers of cultural competence:exploring the meaning of meta cultural competence in the therapeutic community. AB - Social work practice has become more complex and complicated, as social workers wade through the burgeoning call for empirical evidence, the demand to deliver services in shorter periods and with restricted resources. In substance abuse treatment organizations, implementing evidence-based, culturally competent treatment initiatives can seem counter-intuitive to program staff and are subsequently not administered or administered in a way that sabotages the integrity of the intervention. This paper examines cultural competence and explores the meaning of meta-cultural competency that is, the organization's culture, the client's cultural background, and how these factors are interpreted by practitioners within the organizational network. PMID- 17135169 TI - Differences between Black and White elementary school children's orientations toward alcohol and cocaine: a three-study comparison. AB - To trace the origins of race differences in substance use, this study examined differences between Black and White elementary school children's knowledge of alcohol and cocaine, beliefs about their short- and long-term effects, and attitudes toward and intentions to use them across three independent samples (N = 181, N = 287, N = 234). Black children were more negatively oriented toward alcohol and cocaine than White children from an early age. Most notably, in all samples Black children had less positive attitudes toward adult alcohol use and lower intentions to use alcohol. Black children were also more likely to attribute negative long-term health and social effects to alcohol and cocaine use, but there were few significant race differences in knowledge or in expectancies regarding short-term effects of use. Since race differences in beliefs, exposure to alcohol, and socioeconomic factors could not explain race differences in attitudes toward substance use, other cultural differences must be considered. PMID- 17135170 TI - Longitudinal drinking patterns in indigenous Sami and non-indigenous youth in northern Norway. AB - Drinking patterns among indigenous Sami and non-Sami adolescents in northern Norway were explored. Longitudinal data were collected in 1994/95 among 2,950 high school students (RR: 85%) and in 1997/98 among 1,510 follow-ups (RR: 57%). Longitudinally, Sami had significantly lower drinking rates for all measures. An increase in 30-day drinkers occurred during the follow-up period: 23% in Sami and 19% in non-Sami. Sami reported higher paternal abstinence. Sami drank more often in public places. Sami reported more worries from friends and family about their drinking. The lower drinking rates in Sami relative to majority adolescents contrast with findings among indigenous peers elsewhere, but corresponds with the lower parental drinking rates. PMID- 17135171 TI - Retention of African-American and White youth in a longitudinal substance use study. AB - The current investigation discusses successful strategies used to retain N = 405 African-American and White target youth in a longitudinal, non-intervention study focused on alcohol and other drug use. Ninety-one percent of youth remained in the study for all 4 years, including 87% of African-Americans and 96% of Whites. In a logistic regression model incorporating age, ethnicity, income, sex, parent/guardian marital status, parent alcohol use, and family cohesion, only sex significantly predicted retention, with girls being more likely to remain in the study compared with boys, although ethnicity neared significance. PMID- 17135172 TI - High notes: the role of drugs in the making of Jazz. AB - This paper examines the role played by illicit drugs, especially marijuana and heroin, in the historic development and evolution of Jazz in the United States during the twentieth century. In addition to an assessment of the extent of drug use and kinds of drugs used by Jazz musicians and singers, the impact and costs of drug use on the lives of people in Jazz, and the changing patterns of drug use during several eras of Jazz production, the paper contextualizes drug use among Jazz performers and societal response to it in light of prevailing ethnic inequalities and critical medical anthropological theory. PMID- 17135173 TI - A review of perspectives on alcohol and alcoholism in the history of American health and medicine. AB - This review of alcohol use and alcoholism in the history of American health and medicine reveals a range of ambiguous perspectives. In early America, alcohol was attributed with an array of medicinal uses, while habitual drunkenness was not accepted and was identified as a sin. The reformers of the temperance movement expanded upon what they regarded as the social problems associated with alcohol. In nineteenth century America, medical sectarians developed conflicting and contradictory views on health and healing, including the place of alcohol and how to address alcoholism. As the American hospital system evolved, approaches for the care and treatment that alcoholics received in hospitals had to develop as well. Progressive reforms in the early twentieth century impacted many areas of public health in the United States, but continued to embody moralism. These historical trends profoundly influenced the social and institutional responses to alcoholism that continue today, including the evolution of the modern addiction treatment system and the formulation and promulgation of the disease concept. PMID- 17135174 TI - Educational half day: an innovative way to incorporate substance abuse curriculum into residency training. AB - Internal medicine residents often come across substance use disorder (SUD) issues; however not all residency programs have curricula to educate residents on management of these issues. We hereby describe a curricular action plan incorporating an innovative method of Educational Half Day (EHD) to address SUD issues in small group settings. We believe that EHD is an effective way of addressing substance abuse issues in the residency curriculum and it has the potential to change the way residents manage their patients with SUD. PMID- 17135175 TI - Teaching internal medicine resident physicians about Alcoholics Anonymous: a pilot study of an educational intervention. AB - Greater physician confidence in treating alcoholism is associated with a higher frequency of referring alcoholic patients for treatment, but many physicians have limited experience with Alcoholics Anonymous. We implemented a brief, didactic and experiential educational intervention about AA and evaluated its effect on knowledge and attitudes, using a before-after repeated measures study design. Thirty-six first-year internal medicine resident physicians received an educational intervention, which consisted of a 45-minute lecture about AA, a visit to an AA meeting, and a 30-minute debriefing session the next day. Residents' knowledge and attitudes were assessed by a brief written anonymous survey before and after the educational intervention. Residents reported increases in self-perceived knowledge about AA and had more favorable attitudes towards AA after the intervention. Our pilot study shows that a brief, didactic and experiential course can improve physician knowledge and attitudes about AA, and holds promise for improving physician interface with this commonly used intervention. PMID- 17135176 TI - Training HIV physicians to prescribe buprenorphine for opioid dependence. AB - Few HIV physicians are trained to provide buprenorphine treatment. We conducted a cross-sectional survey to assess the impact of an eight-hour course on the treatment of opioid dependence on HIV physicians' preparedness to prescribe buprenorphine. 113 of 257 trained physicians (44%) provided HIV care. Post course, the majority of both HIV physicians and non-HIV physicians (66% vs. 67%, P = .8) planned to pursue a registration to prescribe buprenorphine. The most common reason for not planning to do so was lack of experience (9% vs. 15%, P = .19). 52 of the 113 (46%) HIV physicians had concerns about prescribing buprenorphine. 30 of the 52 (58%) indicated that interactions between buprenorphine and HAART was their primary concern. Following training, most physicians feel prepared and plan to obtain a registration to prescribe buprenorphine. HIV physicians' concerns regarding interactions between buprenorphine and HAART need to be addressed in future training efforts. PMID- 17135177 TI - Technology-based training in cognitive behavioral therapy for substance abuse counselors. AB - This study compared the learning outcomes achieved by 166 practicing substance abuse counselors who were randomized to one of three conditions: (1) a Web-Based Training (WBT) module designed to familiarize practitioners with the "Coping with Craving" module from the NIDA treatment manual, "A Cognitive-Behavioral Approach: Treating Cocaine Addiction" (www.nidatoolbox.org), (2) a face-to-face training workshop covering the identical content, or (3) a delayed training control condition. Participants in all three conditions completed an identical test of knowledge before and after the session. Results indicated that participants in both the WBT and face-to-face workshop conditions showed improvement in mean test scores, while participants in the delayed training control condition did not. Improvements in test scores for participants in both the WBT and face-to-face conditions were statistically significant and equally modest in magnitude. The finding of equivalent rates of knowledge transfer across these two delivery mechanisms highlight one function for which WBT may be particularly well-suited, and suggest that the most effective clinical training applications may use a "blended delivery" format that leverages the strengths of both WBT and face-to face training. PMID- 17135178 TI - Assessment of alcohol and other drug use behaviors in health professions students. AB - Alcohol and other drug (AOD) use behaviors of health professions students (HPS) were assessed by surveying both university-based HPS and other nursing programs in a Midwestern state in 1999. Response was 2,646 (56.4%) of surveyed students. Family history of alcohol-related and drug-related problems were reported by 39.8% and 13.9%, respectively, with 42.6% of respondents reporting one or both. Among nursing respondents, 48.1%, 19.2% and 51.1%, respectively, reported family problems with alcohol, drugs, or one or both. Past-year alcohol use was comparable to undergraduate college students (UCS) nationally (83%); heavy drinking, tobacco and recreational drug use by HPS were lower. Past year drug use was highest among medical students. Marijuana was the predominant illicit drug; medical students and males most often reported use. Health professions educational systems should proactively address student AOD prevention, education and assistance needs. PMID- 17135179 TI - Evaluation of a combined online and in person training in the use of buprenorphine. AB - To evaluate buprenorphine training methodology, we surveyed physicians who had completed a combined online and in person buprenorphine curriculum. Of 53/70 (76%) survey respondents, 57% were psychiatrists and 40% generalists. On a scale of 1 (very poor) to 7 (superlative), the overall training rated a mean of 5.8. The online course (5.0) rated lower than in person training components (p < .001) except for material that addressed the logistics of office practice. The in person patient interview received the highest rating (mean 6.3, p < .001). The 67% of physicians who intended to prescribe buprenorphine after the training were more likely than hesitant physicians to agree that the course provided enough information (p < .05) and that telephone access to experienced providers would improve their confidence (p < .05). Physicians hesitant to prescribe cited lack of experience as the main barrier (41%), with 24% concerned about induction difficulty and reimbursement. Overall, physicians preferred in person instruction and may benefit from additional experiential training and support after curriculum participation. PMID- 17135180 TI - Duration of nonmethadone outpatient treatment: results from a national survey. AB - OBJECTIVES: This study examined organizational trends from 1990 to 2000 and unit characteristics associated with the duration of nonmethadone outpatient addiction treatment. METHODS: Program directors and clinical supervisors from a nationally representative panel of nonmethadone outpatient units in the United States were surveyed in 1990, 1995, and 2000. Treatment duration was measured from clinical supervisors' reports of the average length of stay. Negative binominal regression models controlled for multivariate effects. RESULTS: Treatment duration modestly declined between 1990 and 2000 while addiction severity increased. Affiliation with a mental health center, older program age, JCAHO accreditation-ostensibly a marker for structural quality-and serving more clients with prior authorization requirements- a measure of managed care stringency-were associated with shorter treatment durations. CONCLUSIONS: These findings suggest that treatment duration did not increase between 1990 and 2000 despite clients' worsening addiction severity and growing evidence that longer duration of formal treatment improves treatment outcome. In addition, programs with JCAHO accreditation and stronger managed care oversight appeared to seek efficiencies through reductions in treatment duration. PMID- 17135182 TI - Colorimetric redox-indicator methods for the rapid detection of multidrug resistance in Mycobacterium tuberculosis: a systematic review and meta-analysis. AB - OBJECTIVES: With the spread of multidrug-resistant tuberculosis (MDR-TB) there is increasing demand for new accurate and cost-effective tools for rapid drug susceptibility testing (DST), particularly for developing countries. The reference standard method used today for DST is very slow and cumbersome. Colorimetric assays using redox indicators have been proposed to be used in low resource countries as rapid alternative culture methods for the detection of resistance especially to rifampicin and isoniazid. These methods appear as promising new tools but their accuracy has not been systematically evaluated. METHODS: We did a meta-analysis to evaluate the accuracy of the colorimetric assays for the detection of rifampicin and isoniazid-resistant tuberculosis among clinical isolates. We searched Medline, PubMed (NCBI), Global health-CAB, EJS-E (EbscoHost), ISI Web, Web of Science and IFCC databases and contacted authors if additional information was needed. RESULTS: Eighteen studies met our inclusion criteria for rifampicin resistance detection and 16 for isoniazid. We used a summary receiver operating characteristic (SROC) curve to perform meta-analysis and summarize diagnostic accuracy. For both drugs, all studies had a sensitivity and specificity that ranged between 89% and 100%. CONCLUSIONS: There is evidence that colorimetric methods are highly sensitive and specific for the rapid detection of MDR-TB. These new tools could offer affordable technologies for TB laboratories especially in places where resources are limited and where the prevalence of MDR-TB is important and make TB control efforts more effective. Additional studies are needed in high MDR prevalence countries and cost effectiveness analysis to have more evidence on the utility of these methods. Future developments to detect resistance directly from smear-positive sputum specimens should be taken into consideration to speed up the process. PMID- 17135183 TI - Is continuous infusion ceftriaxone better than once-a-day dosing in intensive care? A randomized controlled pilot study. AB - OBJECTIVES: To compare the clinical and bacteriological outcome of critically ill patients with sepsis treated by ceftriaxone administered as a once-a-day intermittent bolus dose or by 24 h continuous infusion. PATIENTS AND METHODS: We conducted an open-label, randomized controlled pilot study in 57 patients clinically diagnosed with sepsis (suspected/proven infection and systemic inflammatory response syndrome) in a tertiary level intensive care unit. Patients were randomized to receive 2 g of ceftriaxone administered by once-daily intermittent bolus dosing or by 24 h continuous infusion. Clinical and bacteriological outcomes were assessed by blinded clinicians. RESULTS: Fifty seven patients were enrolled in the study, 50 of whom fulfilled the a priori definition of treatment for 4 or more days. The infusion (n = 29) and bolus groups (n = 28) were similar in terms of demographics, although the median age of those receiving the infusion was younger. Intention-to-treat analysis found no statistically significant differences in the primary outcomes for clinical response (P = 0.17), clinical cure [infusion n = 13/29 versus bolus n = 5/28; adjusted odds ratio (AOR) = 3.74; 95% confidence interval (95% CI) = 1.11-12.57; P = 0.06], bacteriological response (P = 0.41) and bacteriological cure (infusion n = 18/29 versus bolus 14/28; AOR = 1.64; 95% CI = 0.57-4.70; P = 0.52). However, logistic regression in patients that complied with the a priori definitions who received ceftriaxone by continuous infusion (AOR = 22.8; 95% CI = 2.24-232.3; P = 0.008) or patients with a low Acute Physiology and Chronic Health Evaluation (APACHE) II score (AOR = 0.70; 95% CI = 0.54-0.91; P = 0.008) were associated with an improved clinical outcome when age and Sepsis Organ Failure Assessment (SOFA) score at time of study entry were controlled for. CONCLUSIONS: This pilot study suggests clinical and bacteriological advantages of continuous infusion of ceftriaxone over bolus administration in critically ill patients in patients requiring 4 or more days of treatment. This sets the scene for a large multicentre double-blind randomized controlled trial to confirm these findings. PMID- 17135184 TI - The interaction networks of structured RNAs. AB - All pairwise interactions occurring between bases which could be detected in three-dimensional structures of crystallized RNA molecules are annotated on new planar diagrams. The diagrams attempt to map the underlying complex networks of base-base interactions and, especially, they aim at conveying key relationships between helical domains: co-axial stacking, bending and all Watson-Crick as well as non-Watson-Crick base pairs. Although such wiring diagrams cannot replace full stereographic images for correct spatial understanding and representation, they reveal structural similarities as well as the conserved patterns and distances between motifs which are present within the interaction networks of folded RNAs of similar or unrelated functions. Finally, the diagrams could help devising methods for meaningfully transforming RNA structures into graphs amenable to network analysis. PMID- 17135185 TI - SNP@Ethnos: a database of ethnically variant single-nucleotide polymorphisms. AB - Inherited genetic variation plays a critical but largely uncharacterized role in human differentiation. The completion of the International HapMap Project makes it possible to identify loci that may cause human differentiation. We have devised an approach to find such ethnically variant single-nucleotide polymorphisms (ESNPs) from the genotype profile of the populations included in the International HapMap database. We selected ESNPs using the nearest shrunken centroid method (NSCM), and performed multiple tests for genetic heterogeneity and frequency spectrum on genes having ESNPs. The function and disease association of the selected SNPs were also annotated. This resulted in the identification of 100 736 SNPs that appeared uniquely in each ethnic group. Of these SNPs, 1009 were within disease-associated genes, and 85 were predicted as damaging using the Sorting Intolerant From Tolerant system. This study resulted in the creation of the SNP@Ethnos database, which is designed to make this type of detailed genetic variation approach available to a wider range of researchers. SNP@Ethnos is a public database of ESNPs with annotation information that currently contains 100 736 ESNPs from 10 138 genes, and can be accessed at http://variome.net and http://bioportal.net/ or directly at http://bioportal.kobic.re.kr/SNPatETHNIC/. PMID- 17135186 TI - Control of carry-over contamination for PCR-based DNA methylation quantification using bisulfite treated DNA. AB - In this study, we adapted the well known uracil DNA glycosylase (UNG) carry-over prevention system for PCR, and applied it to the analysis of DNA methylation based on sodium bisulfite conversion. As sodium bisulfite treatment converts unmethylated cytosine bases into uracil residues, bisulfite treated DNA is sensitive to UNG treatment. Therefore, UNG cannot be used for carry-over prevention of PCR using bisulfite treated template DNA, as not only contaminating products of previous PCR, but also the actual template will be degraded. We modified the bisulfite treatment procedure and generated DNA containing sulfonated uracil residues. Surprisingly, and in contrast to uracil, 6-sulfonyl uracil containing DNA (SafeBis DNA) is resistant to UNG. We showed that the new procedure removes up to 10,000 copies of contaminating PCR product in a closed PCR vessel without significant loss of analytical or clinical sensitivity of the DNA methylation analysis. PMID- 17135187 TI - Mutations in the MutSalpha interaction interface of MLH1 can abolish DNA mismatch repair. AB - MutLalpha, a heterodimer of MLH1 and PMS2, plays a central role in human DNA mismatch repair. It interacts ATP-dependently with the mismatch detector MutSalpha and assembles and controls further repair enzymes. We tested if the interaction of MutLalpha with DNA-bound MutSalpha is impaired by cancer associated mutations in MLH1, and identified one mutation (Ala128Pro) which abolished interaction as well as mismatch repair activity. Further examinations revealed three more residues whose mutation interfered with interaction. Homology modelling of MLH1 showed that all residues clustered in a small accessible surface patch, suggesting that the major interaction interface of MutLalpha for MutSalpha is located on the edge of an extensive beta-sheet that backs the MLH1 ATP binding pocket. Bioinformatic analysis confirmed that this patch corresponds to a conserved potential protein-protein interaction interface which is present in both human MLH1 and its E.coli homologue MutL. MutL could be site-specifically crosslinked to MutS from this patch, confirming that the bacterial MutL-MutS complex is established by the corresponding interface in MutL. This is the first study that identifies the conserved major MutLalpha-MutSalpha interaction interface in MLH1 and demonstrates that mutations in this interface can affect interaction and mismatch repair, and thereby can also contribute to cancer development. PMID- 17135188 TI - Cellular delivery of small interfering RNA by a non-covalently attached cell penetrating peptide: quantitative analysis of uptake and biological effect. AB - Cell-penetrating peptides (CPPs) have evolved as promising new tools to deliver nucleic acids into cells. So far, the majority of these delivery systems require a covalent linkage between carrier and cargo. To exploit the higher flexibility of a non-covalent strategy, we focused on the characterisation of a novel carrier peptide termed MPGalpha, which spontaneously forms complexes with nucleic acids. Using a luciferase-targeted small interfering RNA (siRNA) as cargo, we optimised the conditions for MPGalpha-mediated transfection of mammalian cells. In this system, reporter gene activity could be inhibited up to 90% with an IC50 value in the sub-nanomolar range. As a key issue, we addressed the cellular uptake mechanism of MPGalpha/siRNA complexes applying various approaches. First, transfection of HeLa cells with MPGalpha/siRNA complexes in the presence of several inhibitors of endocytosis showed a significant reduction of the RNA interference (RNAi) effect. Second, confocal laser microscopy revealed a punctual intracellular pattern rather than a diffuse distribution of fluorescently labelled RNA-cargo. These data provide strong evidence of an endocytotic pathway contributing significantly to the uptake of MPGalpha/siRNA complexes. Finally, we quantified the intracellular number of siRNA molecules after MPGalpha-mediated transfection. The amount of siRNA required to induce half maximal RNAi was 10 000 molecules per cell. Together, the combination of methods provided allows for a detailed side by side quantitative analysis of cargo internalisation and related biological effects. Thus, the overall efficiency of a given delivery technique as well as the mechanism of uptake can be assessed. PMID- 17135189 TI - Multiple primer extension by DNA polymerase on a novel plastic DNA array coated with a biocompatible polymer. AB - DNA microarrays are routinely used to monitor gene expression profiling and single nucleotide polymorphisms (SNPs). However, for practically useful high performance, the detection sensitivity is still not adequate, leaving low expression genes undetected. To resolve this issue, we have developed a new plastic S-BIO PrimeSurface with a biocompatible polymer; its surface chemistry offers an extraordinarily stable thermal property for a lack of pre-activated glass slide surface. The oligonucleotides immobilized on this substrate are robust in boiling water and show no significant loss of hybridization activity during dissociation treatment. This allowed us to hybridize the templates, extend the 3' end of the immobilized DNA primers on the S-Bio by DNA polymerase using deoxynucleotidyl triphosphates (dNTP) as extender units, release the templates by denaturalization and use the same templates for a second round of reactions similar to that of the PCR method. By repeating this cycle, the picomolar concentration range of the template oligonucleotide can be detected as stable signals via the incorporation of labeled dUTP into primers. This method of Multiple Primer EXtension (MPEX) could be further extended as an alternative route for producing DNA microarrays for SNP analyses via simple template preparation such as reverse transcript cDNA or restriction enzyme treatment of genome DNA. PMID- 17135190 TI - PeroxisomeDB: a database for the peroxisomal proteome, functional genomics and disease. AB - Peroxisomes are essential organelles of eukaryotic origin, ubiquitously distributed in cells and organisms, playing key roles in lipid and antioxidant metabolism. Loss or malfunction of peroxisomes causes more than 20 fatal inherited conditions. We have created a peroxisomal database (http://www.peroxisomeDB.org) that includes the complete peroxisomal proteome of Homo sapiens and Saccharomyces cerevisiae, by gathering, updating and integrating the available genetic and functional information on peroxisomal genes. PeroxisomeDB is structured in interrelated sections 'Genes', 'Functions', 'Metabolic pathways' and 'Diseases', that include hyperlinks to selected features of NCBI, ENSEMBL and UCSC databases. We have designed graphical depictions of the main peroxisomal metabolic routes and have included updated flow charts for diagnosis. Precomputed BLAST, PSI-BLAST, multiple sequence alignment (MUSCLE) and phylogenetic trees are provided to assist in direct multispecies comparison to study evolutionary conserved functions and pathways. Highlights of the PeroxisomeDB include new tools developed for facilitating (i) identification of novel peroxisomal proteins, by means of identifying proteins carrying peroxisome targeting signal (PTS) motifs, (ii) detection of peroxisomes in silico, particularly useful for screening the deluge of newly sequenced genomes. PeroxisomeDB should contribute to the systematic characterization of the peroxisomal proteome and facilitate system biology approaches on the organelle. PMID- 17135191 TI - FINDbase: a relational database recording frequencies of genetic defects leading to inherited disorders worldwide. AB - Frequency of INherited Disorders database (FINDbase) (http://www.findbase.org) is a relational database, derived from the ETHNOS software, recording frequencies of causative mutations leading to inherited disorders worldwide. Database records include the population and ethnic group, the disorder name and the related gene, accompanied by links to any corresponding locus-specific mutation database, to the respective Online Mendelian Inheritance in Man entries and the mutation together with its frequency in that population. The initial information is derived from the published literature, locus-specific databases and genetic disease consortia. FINDbase offers a user-friendly query interface, providing instant access to the list and frequencies of the different mutations. Query outputs can be either in a table or graphical format, accompanied by reference(s) on the data source. Registered users from three different groups, namely administrator, national coordinator and curator, are responsible for database curation and/or data entry/correction online via a password-protected interface. Databaseaccess is free of charge and there are no registration requirements for data querying. FINDbase provides a simple, web-based system for population-based mutation data collection and retrieval and can serve not only as a valuable online tool for molecular genetic testing of inherited disorders but also as a non-profit model for sustainable database funding, in the form of a 'database journal'. PMID- 17135192 TI - MiST: a microbial signal transduction database. AB - Signal transduction pathways control most cellular activities in living cells ranging from regulation of gene expression to fine-tuning enzymatic activity and controlling motile behavior in response to extracellular and intracellular signals. Because of their extreme sequence variability and extensive domain shuffling, signal transduction proteins are difficult to identify, and their current annotation in most leading databases is often incomplete or erroneous. To overcome this problem, we have developed the microbial signal transduction (MiST) database (http://genomics.ornl.gov/mist), a comprehensive library of the signal transduction proteins from completely sequenced bacterial and archaeal genomes. By searching for domain profiles that implicate a particular protein as participating in signal transduction, we have systematically identified 69 270 two- and one-component proteins in 365 bacterial and archaeal genomes. We have designed a user-friendly website to access and browse the predicted signal transduction proteins within various organisms. Further capabilities include gene/protein sequence retrieval, visualized domain architectures, interactive chromosomal views for exploring gene neighborhood, advanced querying options and cross-species comparison. Newly available, complete genomes are loaded into the database each month. MiST is the only comprehensive and up-to-date electronic catalog of the signaling machinery in microbial genomes. PMID- 17135193 TI - TransportDB: a comprehensive database resource for cytoplasmic membrane transport systems and outer membrane channels. AB - TransportDB (http://www.membranetransport.org/) is a comprehensive database resource of information on cytoplasmic membrane transporters and outer membrane channels in organisms whose complete genome sequences are available. The complete set of membrane transport systems and outer membrane channels of each organism are annotated based on a series of experimental and bioinformatic evidence and classified into different types and families according to their mode of transport, bioenergetics, molecular phylogeny and substrate specificities. User friendly web interfaces are designed for easy access, query and download of the data. Features of the TransportDB website include text-based and BLAST search tools against known transporter and outer membrane channel proteins; comparison of transporter and outer membrane channel contents from different organisms; known 3D structures of transporters, and phylogenetic trees of transporter families. On individual protein pages, users can find detailed functional annotation, supporting bioinformatic evidence, protein/DNA sequences, publications and cross-referenced external online resource links. TransportDB has now been in existence for over 10 years and continues to be regularly updated with new evidence and data from newly sequenced genomes, as well as having new features added periodically. PMID- 17135194 TI - GenomeRNAi: a database for cell-based RNAi phenotypes. AB - RNA interference (RNAi) has emerged as a powerful tool to generate loss-of function phenotypes in a variety of organisms. Combined with the sequence information of almost completely annotated genomes, RNAi technologies have opened new avenues to conduct systematic genetic screens for every annotated gene in the genome. As increasing large datasets of RNAi-induced phenotypes become available, an important challenge remains the systematic integration and annotation of functional information. Genome-wide RNAi screens have been performed both in Caenorhabditis elegans and Drosophila for a variety of phenotypes and several RNAi libraries have become available to assess phenotypes for almost every gene in the genome. These screens were performed using different types of assays from visible phenotypes to focused transcriptional readouts and provide a rich data source for functional annotation across different species. The GenomeRNAi database provides access to published RNAi phenotypes obtained from cell-based screens and maps them to their genomic locus, including possible non-specific regions. The database also gives access to sequence information of RNAi probes used in various screens. It can be searched by phenotype, by gene, by RNAi probe or by sequence and is accessible at http://rnai.dkfz.de. PMID- 17135195 TI - Mouse Tumor Biology Database (MTB): status update and future directions. AB - The Mouse Tumor Biology (MTB) database provides access to data about endogenously arising tumors (both spontaneous and induced) in genetically defined mice (inbred, hybrid, mutant and genetically engineered mice). Data include information on the frequency and latency of mouse tumors, pathology reports and images, genomic changes occurring in the tumors, genetic (strain) background and literature or contributor citations. Data are curated from the primary literature or submitted directly from researchers. MTB is accessed via the Mouse Genome Informatics web site (http://www.informatics.jax.org). Integrated searches of MTB are enabled through use of multiple controlled vocabularies and by adherence to standardized nomenclature, when available. Recently MTB has been redesigned and its database infrastructure replaced with a robust relational database management system (RDMS). Web interface improvements include a new advanced query form and enhancements to already existing search capabilities. The Tumor Frequency Grid has been revised to enhance interactivity, providing an overview of reported tumor incidence across mouse strains and an entree into the database. A new pathology data submission tool allows users to submit, edit and release data to the MTB system. PMID- 17135196 TI - MBGD: a platform for microbial comparative genomics based on the automated construction of orthologous groups. AB - The microbial genome database for comparative analysis (MBGD) is a comprehensive platform for microbial comparative genomics. The central function of MBGD is to create orthologous groups among multiple genomes from precomputed all-against-all similarity relationships using the DomClust algorithm. The database now contains >300 published genomes and the number continues to grow. For researchers who are interested in ongoing genome projects, we have now started a new service called 'My MBGD,' which allows users to add their own genome sequences to MBGD for the purpose of identifying orthologs among both the new and the existing genomes. Furthermore, in order to make available the rapidly accumulating information on closely related genome sequences, we enhanced the interface for pairwise genome comparisons using the CGAT interface, which allows users to see nucleotide sequence alignments of non-coding as well as coding regions. MBGD is available at http://mbgd.genome.ad.jp/. PMID- 17135197 TI - Developments in CORG: a gene-centric comparative genomics resource. AB - The CORG resource (Comparative Regulatory Genomics, http://corg.eb.tuebingen.mpg.de) provides extensive cross-species comparisons of promoter regions in particular and whole gene loci in general. Pairwise as well as multiple alignments of 10 vertebrate species form the key component of CORG. We implemented a rapid alignment approach based on weight matrix motif anchors to ensure efficient computation and biologically informative alignments. All CORG workbench components have been enhanced towards more flexibility and interactivity. Reference sequence based data presentation and analysis was put into the well-known and modular Generic Genome Browser framework. Herein, various plugins facilitate online data analysis and integration with static conservation data. Main emphasis was put on the design of a new JAVA WebStart application for comparative data display. Flexible data import and export options for standard formats complete the provided services. PMID- 17135198 TI - Snap: an integrated SNP annotation platform. AB - Snap (Single Nucleotide Polymorphism Annotation Platform) is a server designed to comprehensively analyze single genes and relationships between genes basing on SNPs in the human genome. The aim of the platform is to facilitate the study of SNP finding and analysis within the framework of medical research. Using a user friendly web interface, genes can be searched by name, description, position, SNP ID or clone name. Several public databases are integrated, including gene information from Ensembl, protein features from Uniprot/SWISS-PROT, Pfam and DAS CBS. Gene relationships are fetched from BIND, MINT, KEGG and are integrated with ortholog data from TreeFam to extend the current interaction networks. Integrated tools for primer-design and mis-splicing analysis have been developed to facilitate experimental analysis of individual genes with focus on their variation. Snap is available at http://snap.humgen.au.dk/ and at http://snap.genomics.org.cn/. PMID- 17135199 TI - DOMINO: a database of domain-peptide interactions. AB - Many protein interactions are mediated by small protein modules binding to short linear peptides. DOMINO (http://mint.bio.uniroma2.it/domino/) is an open-access database comprising more than 3900 annotated experiments describing interactions mediated by protein-interaction domains. DOMINO can be searched with a versatile search tool and the interaction networks can be visualized with a convenient graphic display applet that explicitly identifies the domains/sites involved in the interactions. PMID- 17135200 TI - The CATH domain structure database: new protocols and classification levels give a more comprehensive resource for exploring evolution. AB - We report the latest release (version 3.0) of the CATH protein domain database (http://www.cathdb.info). There has been a 20% increase in the number of structural domains classified in CATH, up to 86 151 domains. Release 3.0 comprises 1110 fold groups and 2147 homologous superfamilies. To cope with the increases in diverse structural homologues being determined by the structural genomics initiatives, more sensitive methods have been developed for identifying boundaries in multi-domain proteins and for recognising homologues. The CATH classification update is now being driven by an integrated pipeline that links these automated procedures with validation steps, that have been made easier by the provision of information rich web pages summarising comparison scores and relevant links to external sites for each domain being classified. An analysis of the population of domains in the CATH hierarchy and several domain characteristics are presented for version 3.0. We also report an update of the CATH Dictionary of homologous structures (CATH-DHS) which now contains multiple structural alignments, consensus information and functional annotations for 1459 well populated superfamilies in CATH. CATH is directly linked to the Gene3D database which is a projection of CATH structural data onto approximately 2 million sequences in completed genomes and UniProt. PMID- 17135201 TI - MMDB: annotating protein sequences with Entrez's 3D-structure database. AB - Three-dimensional (3D) structure is now known for a large fraction of all protein families. Thus, it has become rather likely that one will find a homolog with known 3D structure when searching a sequence database with an arbitrary query sequence. Depending on the extent of similarity, such neighbor relationships may allow one to infer biological function and to identify functional sites such as binding motifs or catalytic centers. Entrez's 3D-structure database, the Molecular Modeling Database (MMDB), provides easy access to the richness of 3D structure data and its large potential for functional annotation. Entrez's search engine offers several tools to assist biologist users: (i) links between databases, such as between protein sequences and structures, (ii) pre-computed sequence and structure neighbors, (iii) visualization of structure and sequence/structure alignment. Here, we describe an annotation service that combines some of these tools automatically, Entrez's 'Related Structure' links. For all proteins in Entrez, similar sequences with known 3D structure are detected by BLAST and alignments are recorded. The 'Related Structure' service summarizes this information and presents 3D views mapping sequence residues onto all 3D structures available in MMDB (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=structure). PMID- 17135202 TI - CDD: a conserved domain database for interactive domain family analysis. AB - The conserved domain database (CDD) is part of NCBI's Entrez database system and serves as a primary resource for the annotation of conserved domain footprints on protein sequences in Entrez. Entrez's global query interface can be accessed at http://www.ncbi.nlm.nih.gov/Entrez and will search CDD and many other databases. Domain annotation for proteins in Entrez has been pre-computed and is readily available in the form of 'Conserved Domain' links. Novel protein sequences can be scanned against CDD using the CD-Search service; this service searches databases of CDD-derived profile models with protein sequence queries using BLAST heuristics, at http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi. Protein query sequences submitted to NCBI's protein BLAST search service are scanned for conserved domain signatures by default. The CDD collection contains models imported from Pfam, SMART and COG, as well as domain models curated at NCBI. NCBI curated models are organized into hierarchies of domains related by common descent. Here we report on the status of the curation effort and present a novel helper application, CDTree, which enables users of the CDD resource to examine curated hierarchies. More importantly, CDD and CDTree used in concert, serve as a powerful tool in protein classification, as they allow users to analyze protein sequences in the context of domain family hierarchies. PMID- 17135203 TI - MINT: the Molecular INTeraction database. AB - The Molecular INTeraction database (MINT, http://mint.bio.uniroma2.it/mint/) aims at storing, in a structured format, information about molecular interactions (MIs) by extracting experimental details from work published in peer-reviewed journals. At present the MINT team focuses the curation work on physical interactions between proteins. Genetic or computationally inferred interactions are not included in the database. Over the past four years MINT has undergone extensive revision. The new version of MINT is based on a completely remodeled database structure, which offers more efficient data exploration and analysis, and is characterized by entries with a richer annotation. Over the past few years the number of curated physical interactions has soared to over 95 000. The whole dataset can be freely accessed online in both interactive and batch modes through web-based interfaces and an FTP server. MINT now includes, as an integrated addition, HomoMINT, a database of interactions between human proteins inferred from experiments with ortholog proteins in model organisms (http://mint.bio.uniroma2.it/mint/). PMID- 17135204 TI - Sentra: a database of signal transduction proteins for comparative genome analysis. AB - Sentra (http://compbio.mcs.anl.gov/sentra), a database of signal transduction proteins encoded in completely sequenced prokaryotic genomes, has been updated to reflect recent advances in understanding signal transduction events on a whole genome scale. Sentra consists of two principal components, a manually curated list of signal transduction proteins in 202 completely sequenced prokaryotic genomes and an automatically generated listing of predicted signaling proteins in 235 sequenced genomes that are awaiting manual curation. In addition to two component histidine kinases and response regulators, the database now lists manually curated Ser/Thr/Tyr protein kinases and protein phosphatases, as well as adenylate and diguanylate cyclases and c-di-GMP phosphodiesterases, as defined in several recent reviews. All entries in Sentra are extensively annotated with relevant information from public databases (e.g. UniProt, KEGG, PDB and NCBI). Sentra's infrastructure was redesigned to support interactive cross-genome comparisons of signal transduction capabilities of prokaryotic organisms from a taxonomic and phenotypic perspective and in the framework of signal transduction pathways from KEGG. Sentra leverages the PUMA2 system to support interactive analysis and annotation of signal transduction proteins by the users. PMID- 17135205 TI - AnimalQTLdb: a livestock QTL database tool set for positional QTL information mining and beyond. AB - The Animal Quantitative Trait Loci (QTL) database (AnimalQTLdb) is designed to house all publicly available QTL data on livestock animal species from which researchers can easily locate and compare QTL within species. The database tools are also added to link the QTL data to other types of genomic information, such as radiation hybrid (RH) maps, finger printed contig (FPC) physical maps, linkage maps and comparative maps to the human genome, etc. Currently, this database contains data on 1287 pig, 630 cattle and 657 chicken QTL, which are dynamically linked to respective RH, FPC and human comparative maps. We plan to apply the tool to other animal species, and add more structural genome information for alignment, in an attempt to aid comparative structural genome studies (http://www.animalgenome.org/QTLdb/). PMID- 17135206 TI - The mouse genome database (MGD): new features facilitating a model system. AB - The mouse genome database (MGD, http://www.informatics.jax.org/), the international community database for mouse, provides access to extensive integrated data on the genetics, genomics and biology of the laboratory mouse. The mouse is an excellent and unique animal surrogate for studying normal development and disease processes in humans. Thus, MGD's primary goals are to facilitate the use of mouse models for studying human disease and enable the development of translational research hypotheses based on comparative genotype, phenotype and functional analyses. Core MGD data content includes gene characterization and functions, phenotype and disease model descriptions, DNA and protein sequence data, polymorphisms, gene mapping data and genome coordinates, and comparative gene data focused on mammals. Data are integrated from diverse sources, ranging from major resource centers to individual investigator laboratories and the scientific literature, using a combination of automated processes and expert human curation. MGD collaborates with the bioinformatics community on the development of data and semantic standards, and it incorporates key ontologies into the MGD annotation system, including the Gene Ontology (GO), the Mammalian Phenotype Ontology, and the Anatomical Dictionary for Mouse Development and the Adult Anatomy. MGD is the authoritative source for mouse nomenclature for genes, alleles, and mouse strains, and for GO annotations to mouse genes. MGD provides a unique platform for data mining and hypothesis generation where one can express complex queries simultaneously addressing phenotypic effects, biochemical function and process, sub-cellular location, expression, sequence, polymorphism and mapping data. Both web-based querying and computational access to data are provided. Recent improvements in MGD described here include the incorporation of single nucleotide polymorphism data and search tools, the addition of PIR gene superfamily classifications, phenotype data for NIH-acquired knockout mice, images for mouse phenotypic genotypes, new functional graph displays of GO annotations, and new orthology displays including sequence information and graphic displays. PMID- 17135207 TI - CellCircuits: a database of protein network models. AB - CellCircuits (http://www.cellcircuits.org) is an open-access database of molecular network models, designed to bridge the gap between databases of individual pairwise molecular interactions and databases of validated pathways. CellCircuits captures the output from an increasing number of approaches that screen molecular interaction networks to identify functional subnetworks, based on their correspondence with expression or phenotypic data, their internal structure or their conservation across species. This initial release catalogs 2019 computationally derived models drawn from 11 journal articles and spanning five organisms (yeast, worm, fly, Plasmodium falciparum and human). Models are available either as images or in machine-readable formats and can be queried by the names of proteins they contain or by their enriched biological functions. We envision CellCircuits as a clearinghouse in which theorists may distribute or revise models in need of validation and experimentalists may search for models or specific hypotheses relevant to their interests. We demonstrate how such a repository of network models is a novel systems biology resource by performing several meta-analyses not currently possible with existing databases. PMID- 17135208 TI - AgBase: a unified resource for functional analysis in agriculture. AB - Analysis of functional genomics (transcriptomics and proteomics) datasets is hindered in agricultural species because agricultural genome sequences have relatively poor structural and functional annotation. To facilitate systems biology in these species we have established the curated, web-accessible, public resource 'AgBase' (www.agbase.msstate.edu). We have improved the structural annotation of agriculturally important genomes by experimentally confirming the in vivo expression of electronically predicted proteins and by proteogenomic mapping. Proteogenomic data are available from the AgBase proteogenomics link. We contribute Gene Ontology (GO) annotations and we provide a two tier system of GO annotations for users. The 'GO Consortium' gene association file contains the most rigorous GO annotations based solely on experimental data. The 'Community' gene association file contains GO annotations based on expert community knowledge (annotations based directly from author statements and submitted annotations from the community) and annotations for predicted proteins. We have developed two tools for proteomics analysis and these are freely available on request. A suite of tools for analyzing functional genomics datasets using the GO is available online at the AgBase site. We encourage and publicly acknowledge GO annotations from researchers and provide an online mechanism for agricultural researchers to submit requests for GO annotations. PMID- 17135209 TI - Structure of human MRG15 chromo domain and its binding to Lys36-methylated histone H3. AB - Human MRG15 is a transcription factor that plays a vital role in embryonic development, cell proliferation and cellular senescence. It comprises a putative chromo domain in the N-terminal part that has been shown to participate in chromatin remodeling and transcription regulation. We report here the crystal structure of human MRG15 chromo domain at 2.2 A resolution. The MRG15 chromo domain consists of a beta-barrel and a long alpha-helix and assumes a structure more similar to the Drosophila MOF chromo barrel domain than the typical HP1/Pc chromo domains. The beta-barrel core contains a hydrophobic pocket formed by three conserved aromatic residues Tyr26, Tyr46 and Trp49 as a potential binding site for a modified residue of histone tail. However, the binding groove for the histone tail seen in the HP1/Pc chromo domains is pre-occupied by an extra beta strand. In vitro binding assay results indicate that the MRG15 chromo domain can bind to methylated Lys36, but not methylated Lys4, Lys9 and Lys27 of histone H3. These data together suggest that the MRG15 chromo domain may function as an adaptor module which can bind to a modified histone H3 in a mode different from that of the HP1/Pc chromo domains. PMID- 17135210 TI - The splicing regulators Tra and Tra2 are unusually potent activators of pre-mRNA splicing. AB - Sexual differentiation in Drosophila is regulated through alternative splicing of doublesex. Female-specific splicing is activated through the activity of splicing enhancer complexes assembled on multiple repeat elements. Each of these repeats serves as a binding platform for the cooperative assembly of a heterotrimeric complex consisting of the SR proteins Tra, Tra2 and 9G8. Using quantitative kinetic analyses, we demonstrate that each component of the enhancer complex is capable of recruiting the spliceosome. Surprisingly, Tra, Tra2 and 9G8 are much stronger splicing activators than other SR protein family members and their activation potential is significantly higher than expected from their serine/arginine content. 9G8 activates splicing not only through its RS domains but also through its RNA-binding domain. The RS domains of Tra and Tra2 are required but not sufficient for efficient complex assembly. Thus, the regulated assembly of the dsx enhancer complexes leads to the generation of an extended activation domain to guarantee the 'all or none' splicing switch that is required during Drosophila sexual differentiation. PMID- 17135211 TI - Greene SCPrimer: a rapid comprehensive tool for designing degenerate primers from multiple sequence alignments. AB - Polymerase chain reaction (PCR) is widely applied in clinical and environmental microbiology. Primer design is key to the development of successful assays and is often performed manually by using multiple nucleic acid alignments. Few public software tools exist that allow comprehensive design of degenerate primers for large groups of related targets based on complex multiple sequence alignments. Here we present a method for designing such primers based on tree building followed by application of a set covering algorithm, and demonstrate its utility in compiling Multiplex PCR primer panels for detection and differentiation of viral pathogens. PMID- 17135212 TI - Exposure to wood dust among carpenters in the construction industry in the Netherlands. AB - Exposure to wood dust was measured among 26 carpenters at 13 building projects. Sampling days were chosen randomly. Individual tasks, based on technology applied and material used during a working day, were sampled separately. From these task based measurements, 8 h time-weighted average concentrations were calculated. Sampling was performed in accordance with a protocol that was developed by the carpentry and furniture industry and which was especially designed for sampling of wood dust. Eight hours time-weighted average exposure to wood dust ranged from 0.8 to 11.6 mg m(-3) with a geometric mean (GM) of 3.3 mg m(-3) and a geometric standard deviation (GSD) of 2.1. The probability of exceedance of the OEL, when comparing the estimated concentrations against the Dutch OEL of 2 mg m(-3), was 75%. The highest exposures were measured during sawing of Cempanel sheets. Task based measurements showed lowest exposures when working outdoors (n = 11, AM = 2.2 mg m(-3)), but even then 5 out of 11 task-based exposures exceeded 2 mg m( 3). Indoors the exposure was 5.2 mg m(-3) (AM, n = 29) and when working both indoors and outdoors exposure was 16.2 mg m(-3) (AM, n = 4). In conclusion, long term average exposure to wood dust among carpenters at construction sites is more than 1.5 times the present occupational exposure limit of 2 mg m(-3). The estimated probability of exceedance of the OEL was 75% and a reduction of exposure with a factor 5 is needed to bring the probability of exceedance below 5%. It is intended to lower the exposure limit to 1 mg m(-3) by 1 January 2007. In that case the probability of exceedance of the OEL is 95% and a reduction of exposure with a factor 10 is needed to bring the probability of exceedance below 5%. This can be achieved by using alternative materials, preparation of building material in workshops equipped with exposure controls, alternative equipment and improved ventilation and good housekeeping. PMID- 17135213 TI - Aniline--a 'historical' contact allergen? Current data from the IVDK and review of the literature. AB - OBJECTIVES: To assess whether aniline should be regarded as potential cause of contact allergy (CA). METHODS: Retrospective analysis of clinical data collected in a CA surveillance network (IVDK, www.ivdk.org) between 01/1992 and 06/2004 and review of pertinent literature. RESULTS: In the above period, 25 of the 1119 patients patch tested with aniline had positive (allergic) reactions; in 24 of these 25 patients, CA to p-phenylenediamine, p-aminoazobenzene or (in one case) another para-amino compound was additionally diagnosed. Exposure to aniline could not be ascertained in any of the cases, based on the available data. DISCUSSION: Previous clinical results, which have been summarized and tabulated, are partly difficult to evaluate, as they may lack detail, or test concentrations are higher than those currently recommended, possibly yielding false-positive reactions. In none of the studies had previous exposures to aniline been unequivocally identified. CONCLUSION: Based on clinical data it is unlikely that aniline is an independent sensitizer of current importance. However, it may elicit allergic reactions in subjects pre-sensitized to para-substituted amino compounds. In summary, supported by recent experimental evidence employing the local lymph node assay as a validated animal test system, it appears probable that aniline is a weak allergen. PMID- 17135214 TI - Porous microstructures, obtained by oxidation of (In,Ga)2Se3 single crystals. AB - Surface topology of thermally oxidized (In,Ga)(2)Se(3) single crystals is discussed. It was established that own oxide surfaces possess morphologies of micropores network. The sizes of pores are in agreement of exponential empirical correlation versus oxidation temperature. PMID- 17135215 TI - Development of a specimen heating holder with an evaporator and gas injector and its application for catalyst. AB - A specimen heating holder equipped with a gas injector and an evaporator has been developed for use with conventional transmission electron microscopes (TEMs). The developed specimen holder allows both synthesis of metal oxide support and deposition of catalyst nano-particles in situ. Since the holder is designed to be used in small gapped high-resolution objective lens pole-piece, all the procedure from the synthesis of support material to the deposition of catalyst as well as the behavior of the catalyst nano-particles on the support can be observed at near atomic resolution. The developed specimen holder was applied to the study of AuPd catalyst. First, air was injected onto heated aluminum particles via a gas injector to synthesize Al(2)O(3) support. Then, nano-particles of AuPd were deposited on the Al(2)O(3) support. After the deposition, the synthesized Al(2)O(3) support was heated and air was injected again to observe behaviors of the deposited AuPd nano-particles at elevated temperatures in the aerial environment. Behaviors of the AuPd nano-particles such as coalescence, segmentation and diffusion to the Al(2)O(3) support were dynamically observed at atomic level high resolution. PMID- 17135216 TI - Acute acalculous cholecystitis associated with malarial infection in children: report of two cases. AB - Acute acalculous cholecystitis (AAC) is a serious condition that has rarely been reported in association with malaria. It is particularly rare in pediatric malaria patients and only one case has been reported till date. We report AAC in association with falciparum and vivax malaria in one pediatric patient each. PMID- 17135217 TI - A prospective study of anti-chromatin and anti-C1q autoantibodies in patients with proliferative lupus nephritis treated with cyclophosphamide pulses or azathioprine/methylprednisolone. AB - OBJECTIVE: To study the prevalence and course of anti-chromatin (anti-nucleosome, anti-double-stranded (ds) DNA and anti-histone) and anti-C1q autoantibodies in patients with proliferative lupus nephritis (LN), treated in a randomised controlled trial with either cyclophosphamide or azathioprine plus methylprednisolone. METHODS: Autoantibody levels were measured and analysed in 52 patients with proliferative LN, during their first year of treatment. Levels in both treatment arms were compared and associations with clinical, serological and outcome parameters were studied. RESULTS: At study entry, prevalences for anti nucleosome, anti-dsDNA, anti-histone and anti-C1q autoantibodies were 81%, 96%, 23% and 65%, respectively. Anti-chromatin autoantibodies correlated with each other, but not with anti-C1q levels. If patients were divided for their autoantibody titre at the start of treatment above or below the median, the only significant differences were higher SLE disease activity index with higher anti nucleosome, and higher creatinine with higher anti-C1q autoantibodies. During the first year, a comparable rapid decline in the levels of anti-nucleosome, anti dsDNA and anti-C1q autoantibodies was seen in both treatment arms. Anti-histone autoantibody levels were low and did not change. Renal flares were not preceded by rises in autoantibody titres. CONCLUSIONS: These results indicate that measurement of anti-chromatin and anti-C1q autoantibodies is useful for diagnosing LN, but not for monitoring disease course. PMID- 17135218 TI - Can atrial fibrillation with a coarse electrocardiographic appearance be treated with catheter ablation of the tricuspid valve-inferior vena cava isthmus? Results of a multicentre randomised controlled trial. AB - OBJECTIVE: To see if strategy of ablating the tricuspid annulus-inferior vena cava isthmus (TV-IVC) is superior to electrical cardioversion to prevent recurrences in patients with coarse atrial fibrillation. DESIGN: Prospective randomised controlled multicentre study. SETTING: Four tertiary referral hospitals in the UK. PATIENTS: 57 patients with persistent coarse atrial fibrillation (irregular P waves > or =0.15 mV in > or =1 ECG lead). INTERVENTIONS: Patients were randomised to receive external cardioversion (group A, n = 30) or TV-IVC ablation +/- DC cardioversion (group B, n = 27). MAIN OUTCOME MEASURES: Cardiac rhythm, scores on quality of life and symptom questionnaires were assessed at 4, 16 and 52 weeks after the procedure. RESULTS: 20 (67%) patients in group A and 19 (70%) patients in group B were in sinus rhythm immediately after their index procedure. At 4, 16 and 52 weeks, the number of patients in sinus rhythm were 5, 3 and 2 in group A and 3, 3 and 1 in group B (p = NS). The quality of life and symptom questionnaire scores were similar in the two groups at each period of follow-up, although they were significantly better for sinus rhythm than for atrial fibrillation at each follow-up visit. CONCLUSIONS: As a first-line strategy, TV-IVC ablation offers no advantages over direct current cardioversion for the management of coarse atrial fibrillation. PMID- 17135219 TI - Ventricular size and function assessed by cardiac MRI predict major adverse clinical outcomes late after tetralogy of Fallot repair. AB - BACKGROUND: Factors associated with impaired clinical status in a cross-sectional study of patients with repaired tetralogy of Fallot (TOF) have been reported previously. OBJECTIVES: To determine independent predictors of major adverse clinical outcomes late after TOF repair in the same cohort during follow-up evaluated by cardiac magnetic resonance (CMR). METHODS: Clinical status at latest follow-up was ascertained in 88 patients (median time from TOF repair to baseline evaluation 20.7 years; median follow-up from baseline evaluation to most recent follow-up 4.2 years). Major adverse outcomes included (a) death; (b) sustained ventricular tachycardia; and (c) increase in NYHA class to grade III or IV. RESULTS: 22 major adverse outcomes occurred in 18 patients (20.5%): death in 4, sustained ventricular tachycardia in 8, and increase in NYHA class in 10. Multivariate analysis identified right ventricular (RV) end-diastolic volume Z >or=7 (odds ratio (OR) = 4.55, 95% confidence interval (CI) 1.10 to 18.8, p = 0.037) and left ventricular (LV) ejection fraction <55% (OR = 8.05, 95% CI 2.14 to 30.2, p = 0.002) as independent predictors of outcome with an area under the receiver operator characteristic curve of 0.850. LV ejection fraction could be replaced by RV ejection fraction <45% in the multivariate model. QRS duration >or=180 ms also predicted major adverse events but correlated with RV size. CONCLUSIONS: In this cohort, severe RV dilatation and either LV or RV dysfunction assessed by CMR predicted major adverse clinical events. This information may guide risk stratification and therapeutic interventions. PMID- 17135220 TI - Long term effects of bosentan treatment in adult patients with pulmonary arterial hypertension related to congenital heart disease (Eisenmenger physiology): safety, tolerability, clinical, and haemodynamic effect. AB - BACKGROUND: Oral bosentan is an established treatment for pulmonary arterial hypertension (PAH). OBJECTIVE: To evaluate safety, tolerability, and clinical and haemodynamic effects of bosentan in patients with PAH related to congenital heart disease (CHD). PATIENTS: 22 patients with CHD related PAH (8 men, 14 women, mean (SD) age 38 (10) years) were treated with oral bosentan (62.5 mg x 2/day for the first 4 weeks and then 125 mg x 2/day). MAIN OUTCOME MEASURES: Clinical status, liver enzymes, World Health Organisation (WHO) functional class, resting oxygen saturations and 6-min walk test (6MWT) were assessed at baseline and at 1, 3, 6, and 12 months. Haemodynamic evaluation with cardiac catheterisation was performed at baseline and at 12 month follow-up. RESULTS: 12 patients had ventricular septal defect, 5 atrioventricular canal, 4 single ventricle, and 1 atrial septal defect. All patients tolerated bosentan well. No major side effects were seen. After a year of treatment, an improvement was seen in WHO functional class (2.5 (0.7) v 3.1 (0.7); p<0.05), oxygen saturation at rest (87 (6%) v 81 (9); p<0.001), heart rate at rest (81 (10) v 87 (14) bpm; p<0.05), distance travelled in the 6MWT (394 (73) v 320 (108) m; p<0.001), oxygen saturation at the end of the 6MWT (71 (14) v 63 (17%); p<0.05), Borg index (5.3 (1.8) v 6.5 (1.3); p<0.001), pulmonary vascular resistances index (14 (9) v 22 (12) WU m(2); p<0.001), systemic vascular resistances index (23 (11) v 27 (10) WU.m(2); p<0.01), pulmonary vascular resistances index/systemic vascular resistances index (0.6 (0.5) v 0.9 (0.6); p<0.05); pulmonary (4.0 (1.3) v 2.8 (0.9) l/min/m2; p<0.001) and systemic cardiac output (4.2 (1.4) v 3.4 (1.1) l/min/m2; p<0.05). CONCLUSIONS: Bosentan was safe and well tolerated in adults with CHD related PAH during 12 months of treatment. Clinical status, exercise tolerance, and pulmonary haemodynamics improved considerably. PMID- 17135221 TI - Coronary artery flow velocity profile measured by transthoracic Doppler echocardiography predicts myocardial viability after acute myocardial infarction. AB - OBJECTIVE: To study whether flow velocity profile in the left anterior descending coronary artery (LAD) measured by transthoracic Doppler echocardiography (TTDE) predicts myocardial viability after reperfused anterior acute myocardial infarction (AMI). PATIENTS AND METHODS: 15 patients who had their first anterior ST elevation AMI and were successfully reperfused by coronary angioplasty and five controls without coronary artery disease were selected. Blood flow velocity spectrum was measured from the mid-LAD by TTDE 3 days after coronary angioplasty. Myocardial viability in the LAD region was quantified 3 months after AMI by relative uptake of 18F-fluorodeoxyglucose (FDG) imaged with positron emission tomography. Myocardium was graded as viable, partially viable or non-viable (relative FDG uptake >85%, 67-85% and <67%, respectively). Main outcome measures were diastolic deceleration time (DDT) of LAD flow velocity 3 days after AMI and myocardial viability 3 months after AMI. RESULTS: DDT of LAD flow velocity correlated with myocardial FDG uptake in the LAD region (r = 0.91, p<0.01). DDT was markedly longer in patients with viable myocardium (876+/-76 ms, n = 3) than partially viable (356+/-89 ms, n = 6, p<0.01), or non-viable myocardium (128+/-13 ms, n = 6, p<0.01). In controls, DDT was comparable (909+/-76 ms, n = 5) to patients with viable myocardium. DDT <190 ms was always associated with non viable myocardium. CONCLUSIONS: DDT of LAD flow velocity is strongly associated with myocardial viability after reperfused anterior AMI. Non-invasive TTDE of the LAD may be used in the acute phase to predict long-term viability of the jeopardised myocardium. PMID- 17135222 TI - Impact of blood pressure on the Doppler echocardiographic assessment of severity of aortic stenosis. AB - OBJECTIVE: To investigate the impact of blood pressure (BP) on the Doppler echocardiographic (Doppler-echo) evaluation of severity of aortic stenosis (AS). METHODS: Handgrip exercise or phenylephrine infusion was used to increase BP in 22 patients with AS. Indices of AS severity (mean pressure gradient (DeltaP(mean)), aortic valve area (AVA), valve resistance, percentage left ventricular stroke work loss (% LVSW loss) and the energy loss coefficient (ELCo)) were measured at baseline, peak BP intervention and recovery. RESULTS: From baseline to peak intervention, mean (SD) BP increased (99 (8) vs 121 (10) mm Hg, p<0.001), systemic vascular resistance (SVR) increased (1294 (264) vs 1552 (372) dynexs/cm(5), p<0.001) and mean (SD) transvalvular flow rate (Q(mean)) decreased (323 (67) vs 306 (66) ml/s, p = 0.02). There was no change in DeltaP(mean) (36 (13) vs 36 (14) mm Hg, p = NS). However, there was a decrease in AVA (1.15 (0.32) vs 1.09 (0.33) cm(2), p = 0.02) and ELCo (1.32 (0.40) vs 1.24 (0.42) cm(2), p = 0.04), and an increase in valve resistance (153 (63) vs 164 (74) dynexs/cm(5), p = 0.02), suggesting a more severe valve stenosis. In contrast, % LVSW loss decreased (19.8 (6) vs 16.5 (6)%, p<0.001), suggesting a less severe valve stenosis. There was an inverse relationship between the change in mean BP and AVA (r = -0.34, p = 0.02); however, only the change in Q(mean) was an independent predictor of the change in AVA (r = 0.81, p<0.001). CONCLUSIONS: Acute BP elevation due to increased SVR can affect the Doppler-echo evaluation of AS severity. However, the impact of BP on the assessment of AS severity depends primarily on the associated change in Q(mean), rather than on an independent effect of SVR or arterial compliance, and can result in a valve appearing either more or less stenotic depending on the direction and magnitude of the change in Q(mean). PMID- 17135223 TI - Cost effectiveness of perindopril in reducing cardiovascular events in patients with stable coronary artery disease using data from the EUROPA study. AB - BACKGROUND: The EUropean trial on Reduction Of cardiac events with Perindopril in stable coronary Artery disease (EUROPA) trial has recently reported. OBJECTIVE: To assess the cost effectiveness of perindopril in stable coronary heart disease in the UK. METHODS: Clinical and resource use data were taken from the EUROPA trial. Costs included drugs and hospitalisations. Health-related quality of life values were taken from published sources. A cost-effectiveness analysis is presented as a function of the risk of a primary event (non-fatal myocardial infarction, cardiac arrest or cardiovascular death) in order to identify people for whom treatment offers greatest value for money. RESULTS: The median incremental cost of perindopril for each quality-adjusted life year (QALY) gained across the heterogeneous population of EUROPA was estimated as 9700 pounds(interquartile range 6400-14,200 pounds). Overall, 88% of the EUROPA population had an estimated cost per QALY below 20,000 pounds and 97% below 30,000 pounds. For a threshold value of cost effectiveness of 30,000 pounds per QALY gained, treatment of people representing the 25th, 50th (median) and 75th centiles of the cost effectiveness distribution for perindopril has a probability of 0.999, 0.99 and 0.93 of being cost effective, respectively. Cost effectiveness was strongly related to higher risk of a primary event under standard care. CONCLUSIONS: Whether the use of perindopril can be considered cost effective depends on the threshold value of cost effectiveness of healthcare systems. For the large majority of patients included in EUROPA, the incremental cost per QALY gained was lower than the apparent threshold used by the National Institute for Health and Clinical Excellence in the UK. PMID- 17135224 TI - Left ventricular non-compaction cardiomyopathy in children: characterisation of clinical status using tissue Doppler-derived indices of left ventricular diastolic relaxation. AB - BACKGROUND: Left ventricular non-compaction (LVNC) may manifest an undulating phenotype ranging from dilated to hypertrophic appearance. It is unknown whether tissue Doppler (TD) velocities can predict adverse clinical outcomes including death and need for transplantation in children with LVNC. METHODS AND RESULTS: 56 children (median age 4.5 years, median follow-up 26 months) with LVNC evaluated at one hospital from January 1999 to May 2004 were compared with 56 age/sex matched controls. Children with LVNC had significantly decreased early diastolic TD velocities (Ea) at the lateral mitral (11.0 vs 17.0 cm/s) and septal (8.9 vs 11.0 cm/s) annuli compared with normal controls (p<0.001 for each comparison). Using receiver operator characteristic curves, the lateral mitral Ea velocity proved the most sensitive and specific predictor for meeting the primary end point (PEP) at 1 year after diagnosis (area under the curve = 0.888, SE = 0.048, 95% CI 0.775 to 0.956). A lateral mitral Ea cut-off velocity of 7.8 cm/s had a sensitivity of 87% and a specificity of 79% for the PEP. Freedom from death or transplantation was 85% at 1 year and 77% at 2 years. CONCLUSIONS: TD velocities are significantly reduced in patients with LVNC compared with normal controls. Reduced lateral mitral Ea velocity helps predict children with LVNC who are at risk of adverse clinical outcomes including death and need for cardiac transplantation. PMID- 17135225 TI - The 620W allele is the PTPN22 genetic variant conferring susceptibility to RA in a Dutch population. AB - OBJECTIVES: A missense SNP, C1858T, in PTPN22 has been identified as a genetic risk factor for rheumatoid arthritis (RA). Subsequent work has suggested that other variants in this gene, in particular a haplotype marked by the minor allele of rs3789604, are associated with RA in white North Americans independent of C1858T. We tested this hypothesis in an independent white Dutch study. METHODS: A total of 667 RA patients and 286 controls were genotyped for 13 PTPN22 single nucleotide polymorphisms (SNPs) by allele-specific kinetic polymerase chain reaction. rs3789604 was genotyped in an additional 410 RA and 270 UA patients participating in the Leiden early arthritis inception cohort. We conducted single marker and haplotype association tests. RESULTS: The sole haplotype strongly associated with RA in our Dutch population carries the PTPN22 1858T allele. A second haplotype identical at all other SNPs tested except 1858 was not associated with disease. No significant association of the haplotype tagged by the 3' PTPN22 SNP, rs3789604, with RA susceptibility (P = 0.134) was observed in our sample set. CONCLUSION: We conclude that C1858T is the sole PTPN22 variant predisposing to RA in our white Dutch sample set. PMID- 17135226 TI - Chimerism in systemic lupus erythematosus--three hypotheses. AB - Systemic lupus erythematosus (SLE) is an immune-mediated disease characterized by the presence of autoantibodies and a wide array of clinical symptoms. Despite intensive research, the aetiology of SLE is still unknown and is probably multifactorial. Both genetic and environmental factors have been associated with SLE, but these factors alone are insufficient to explain the onset of SLE. Recently, it has been suggested that chimerism plays a role in the pathogenesis of autoimmune diseases, including SLE. Chimerism indicates the presence of cells from one individual in another individual. In an experimental mouse model, the injection of chimeric cells induces a lupus-like disease. In addition, chimerism is found more often in kidneys of women with SLE than in healthy controls. There are several mechanisms by which chimeric cells could be involved in the pathogenesis of SLE. In this review, three hypotheses on the role of chimerism in SLE are discussed. The first two hypotheses describe the possibilities that chimeric cells induce either a graft-vs-host reaction in the host (comparable with reactions seen after bone marrow transplantation) or a host-vs-graft reaction (comparable with reactions seen after solid organ transplantation). The third hypothesis discusses the possible beneficial role chimeric cells may play in repair mechanisms due to their stem cell-like properties. This review provides insights into the mechanisms by which chimerism may be involved in SLE and proposes several lines of inquiry to further investigate chimerism in SLE. PMID- 17135227 TI - Development of a competency framework for general practitioners with a special interest in musculoskeletal/rheumatology practice. PMID- 17135228 TI - NT-proBNP can be used to detect right ventricular systolic dysfunction in pulmonary hypertension. AB - Right ventricular systolic dysfunction (RVSD) at baseline (pre-treatment) predicts early death in patients with pulmonary hypertension (PH). However, RVSD can only be detected reliably by prohibitively invasive or expensive techniques. N-terminal B-type natriuretic peptide concentration ([NT-proBNP]) correlates with RV function in PH; however, an [NT-proBNP] threshold that indicates RVSD in individual patients has not previously been determined. Twenty-five patients with PH (pulmonary arterial hypertension (n = 19) or chronic thromboembolic PH (n = 6)) underwent cardiovascular magnetic resonance (CMR) imaging and NT-proBNP measurement at baseline. [NT-proBNP] was correlated against RV dimensions and ejection fraction (RVEF) measured directly by CMR imaging. The ability of NT proBNP to detect RVSD (defined as a CMR-derived RVEF >2 SDS below control values) was tested and predictors of [NT-proBNP] identified. [NT-proBNP] correlated negatively with RVEF. RVSD was present in nine out of 25 patients. An [NT-proBNP] threshold of 1,685 pg.mL(-1) was sensitive (100%) and specific (94%) in detecting RVSD. RVEF and RV mass index independently predicted [NT-proBNP]. In pulmonary hypertension, a baseline N-terminal B-type natriuretic peptide concentration of >1,685 ng.L(-1) suggests right ventricular systolic dysfunction, and thus an increased risk of early death. N-terminal B-type natriuretic peptide could prove useful as an objective, noninvasive means of identifying patients with pulmonary hypertension who have right ventricular systolic dysfunction at presentation. PMID- 17135229 TI - Treatment failure in tuberculosis. AB - Treatment of latent tuberculosis (TB) infection with 3 months of rifampicin/isoniazid is a major part of preventive TB programmes. The effectiveness of treatment of latent TB infection can only be assessed by rates of subsequent breakdown and there are few outcome data for this combination of rifampicin/isoniazid. Therefore, the aim of the present study was to estimate the failure rate following treatment for the latent TB infection. A questionnaire survey was carried out in all parents of children aged <16 yrs who completed treatment for latent TB infection at Leicester Royal Infirmary (Leicester, UK) over the period 1997-2003. Cases of treatment failure were identified by reviewing all re-referrals to the clinic, identifying children developing TB while on treatment and by postal questionnaire to all patients discharged. Of the 400 eligible children, 344 (86%) replied. Three children who had latent TB infection subsequently developed TB disease over the time period. Of those three patients, one developed chest radiograph signs at the end of treatment and two presented with symptoms within 2 yrs of completing treatment. Overall, the mean treatment failure rate was 0.87% (95% confidence interval 0.3-2.5) or 2.2 cases per 1,000 patient-yrs. In conclusion, rates of tuberculosis breakdown after treatment for latent tuberculosis infection with 3 months rifampicin/isoniazid are acceptably low. PMID- 17135230 TI - Respiratory heat and moisture loss is associated with eosinophilic inflammation in asthma. AB - Increased mucosal vascularity is a hallmark of airway inflammation in asthma. It was hypothesised that this would lead to a detectable increase in respiratory heat and moisture loss (RHML), which would reflect the degree of airway inflammation present. A total of 23 subjects with asthma and 18 healthy controls had RHML measured in a cross-sectional study. The measurements were made using a device that combines temperature and humidity measurement during inspiration and expiration and allows precise control over inspirate conditions and ventilatory pattern. The subjects with asthma underwent parallel measurements of exhaled nitric oxide, sputum eosinophil percentage and exhaled breath condensate pH. Mean+/-SD RHML was elevated in patients with asthma (98.1+/-7.3 J.L(-1)) compared with control subjects (91.9+/-4.5 J.L(-1)). RHML measurement in asthma correlated with sputum eosinophil percentage. This novel correlation between thermal and cellular measurements in asthma suggests that both of these noninvasive indices are sensitive to the degree of underlying chronic airway inflammation. PMID- 17135231 TI - Effect of indacaterol, a novel long-acting beta2-agonist, on isolated human bronchi. AB - Indacaterol is a novel beta2-adrenoceptor agonist in development for the once daily treatment of asthma and chronic obstructive pulmonary disease. The present study evaluated the relaxant effect of indacaterol on isolated human bronchi obtained from lungs of patients undergoing surgery for lung carcinoma. Potency ( logEC50), maximal relaxant effect (Emax) and onset of action were determined at resting tone. Duration of action was determined against cholinergic neural contraction induced by electrical field stimulation (EFS). At resting tone, logEC50 and Emax values were 8.82+/-0.41 and 77+/-5% for indacaterol, 9.84+/-0.22 and 94+/-1% for formoterol, 8.36+/-0.16 and 74+/-4% for salmeterol, and 8.43+/ 0.22 and 84+/-4% for salbutamol, respectively. In contrast to salmeterol, indacaterol did not antagonise the isoprenaline response. Indacaterol's onset of action (7.8+/-0.7 min) was not significantly different from that of formoterol (5.8+/-0.7 min) or salbutamol (11.0+/-4.0 min), but it was significantly faster than that of salmeterol (19.4+/-4.3 min). EFS-induced contractions were inhibited with -logIC50 values of 6.96+/-0.13 (indacaterol), 8.96+/-0.18 (formoterol), 7.18+/-0.34 (salmeterol) and 6.39+/-0.26 (salbutamol). Duration of action was >12 h for indacaterol and salmeterol, and 35.3+/-8.8 and 14.6+/-3.7 min for formoterol and salbutamol, respectively. In isolated human bronchi, indacaterol behaved as a long-acting beta2-adrenoceptor agonist with high intrinsic efficacy and fast onset of action. PMID- 17135232 TI - Pulmonary artery distensibility in pulmonary arterial hypertension: an MRI pilot study. AB - Pulmonary arterial hypertension (PAH) is a disease of the small vessels in which there is a substantial increase in pulmonary vascular resistance leading to right ventricle failure and death. Invasive haemodynamic evaluation is mandatory not only for diagnosis confirmation but also to address prognosis and eligibility for the use of calcium-channel blockers through an acute vasodilator challenge. Noninvasive surrogate response markers to the acute vasodilator test have been sought. In the present study, the relationship between pulmonary artery distensibility, assessed using magnetic resonance imaging (MRI), and response to acute vasodilator tests was investigated. In total, 19 patients diagnosed with idiopathic PAH without any specific treatment were evaluated. Within a 48-h window after pulmonary artery catheterisation, patients underwent cardiac MRI. Cardiac index, calculated after the determination (invasively and noninvasively) of cardiac output, showed excellent correlation, as did right atrial pressure and right ventricle ejection fraction. Pulmonary artery distensibility was significantly higher in responders. A receiver operating characteristic curve analysis has shown that 10% distensibility was able to differentiate responders from nonresponders with 100% sensitivity and 56% specificity. The present findings suggest that magnetic resonance imaging and pulmonary artery distensibility may be useful noninvasive tools for the evaluation of patients with pulmonary hypertension. PMID- 17135233 TI - Changes in HRCT findings in patients with respiratory bronchiolitis-associated interstitial lung disease after smoking cessation. AB - High-resolution computed tomography (HRCT) findings in patients with respiratory bronchiolitis-associated interstitial lung disease (RB-ILD) are varied and nonspecific. There is no known report of changes in HRCT findings and respiratory function test results for RB-ILD patients following the cessation of smoking. Five patients with RB-ILD, confirmed by surgical lung biopsy, were retrospectively studied. Each stopped cigarette smoking and did not receive corticosteroid therapy after diagnosis. The clinical symptoms, respiratory function test results and HRCT findings obtained at the final observation were compared with those from the time of diagnosis. Ground-glass opacity and centrilobular nodules corresponding to pathological respiratory bronchiolitis, as well as intralobular fine linear-reticular opacity corresponding to fibrosis involving the subpleural alveolar septa, showed computed tomography-pathological correlations. Both clinical symptoms and the diffusing capacity of the lungs for carbon monoxide improved significantly following smoking cessation, as did ground glass opacity and centrilobular nodules seen during the initial HRCT examination. Centrilobular nodules and ground-glass opacity, which are the main features of high-resolution computed tomography of respiratory bronchiolitis-associated interstitial lung disease patients and represent pathological respiratory bronchiolitis, can be improved by smoking cessation. The diffusing capacity of the lung for carbon monoxide in respiratory function tests can be also improved. PMID- 17135234 TI - Protein kinase Cdelta plays a non-redundant role in insulin secretion in pancreatic beta cells. AB - Protein kinase C (PKC) is considered to modulate glucose-stimulated insulin secretion. Pancreatic beta cells express multiple isoforms of PKCs; however, the role of each isoform in glucose-stimulated insulin secretion remains controversial. In this study we investigated the role of PKCdelta, a major isoform expressed in pancreatic beta cells on beta cell function. Here, we showed that PKCdelta null mice manifested glucose intolerance with impaired insulin secretion. Insulin tolerance test showed no decrease in insulin sensitivity in PKCdelta null mice. Studies using islets isolated from these mice demonstrated decreased glucose- and KCl-stimulated insulin secretion. Perifusion studies indicated that mainly the second phase of insulin secretion was decreased. On the other hand, glucose-induced influx of Ca2+ into beta cells was not altered. Immunohistochemistry using total internal reflection fluorescence microscopy and electron microscopic analysis showed an increased number of insulin granules close to the plasma membrane in beta cells of PKCdelta null mice. Although PKC is thought to phosphorylate Munc18-1 and facilitate soluble N-ethylmaleimide sensitive fusion protein attachment protein receptors complex formation, the phosphorylation of Munc18-1 by glucose stimulation was decreased in islets of PKCdelta null mice. We conclude that PKCdelta plays a non-redundant role in glucose-stimulated insulin secretion. The impaired insulin secretion in PKCdelta null mice is associated with reduced phosphorylation of Munc18-1. PMID- 17135235 TI - Knock-out of the magnesium protoporphyrin IX methyltransferase gene in Arabidopsis. Effects on chloroplast development and on chloroplast-to-nucleus signaling. AB - Protoporphyrin IX is the last common intermediate between the heme and chlorophyll biosynthesis pathways. The addition of magnesium directs this molecule toward chlorophyll biosynthesis. The first step downstream from the branchpoint is catalyzed by the magnesium chelatase and is a highly regulated process. The corresponding product, magnesium protoporphyrin IX, has been proposed to play an important role as a signaling molecule implicated in plastid to-nucleus communication. To get more information on the chlorophyll biosynthesis pathway and on magnesium protoporphyrin IX derivative functions, we have identified an magnesium protoporphyrin IX methyltransferase (CHLM) knock-out mutant in Arabidopsis in which the mutation induces a blockage downstream from magnesium protoporphyrin IX and an accumulation of this chlorophyll biosynthesis intermediate. Our results demonstrate that the CHLM gene is essential for the formation of chlorophyll and subsequently for the formation of photosystems I and II and cytochrome b6f complexes. Analysis of gene expression in the chlm mutant provides an independent indication that magnesium protoporphyrin IX is a negative effector of nuclear photosynthetic gene expression, as previously reported. Moreover, it suggests the possible implication of magnesium protoporphyrin IX methyl ester, the product of CHLM, in chloroplast-to-nucleus signaling. Finally, post-transcriptional up-regulation of the level of the CHLH subunit of the magnesium chelatase has been detected in the chlm mutant and most likely corresponds to specific accumulation of this protein inside plastids. This result suggests that the CHLH subunit might play an important regulatory role when the chlorophyll biosynthetic pathway is disrupted at this particular step. PMID- 17135236 TI - Crystal structure of 1-deoxy-D-xylulose 5-phosphate synthase, a crucial enzyme for isoprenoids biosynthesis. AB - Isopentenyl pyrophosphate (IPP) is a common precursor for the synthesis of all isoprenoids, which have important functions in living organisms. IPP is produced by the mevalonate pathway in archaea, fungi, and animals. In contrast, IPP is synthesized by a mevalonate-independent pathway in most bacteria, algae, and plant plastids. 1-Deoxy-D-xylulose 5-phosphate synthase (DXS) catalyzes the first and the rate-limiting step of the mevalonate-independent pathway and is an attractive target for the development of novel antibiotics, antimalarials, and herbicides. We report here the first structural information on DXS, from Escherichia coli and Deinococcus radiodurans, in complex with the coenzyme thiamine pyrophosphate (TPP). The structure contains three domains (I, II, and III), each of which bears homology to the equivalent domains in transketolase and the E1 subunit of pyruvate dehydrogenase. However, DXS has a novel arrangement of these domains as compared with the other enzymes, such that the active site of DXS is located at the interface of domains I and II in the same monomer, whereas that of transketolase is located at the interface of the dimer. The coenzyme TPP is mostly buried in the complex, but the C-2 atom of its thiazolium ring is exposed to a pocket that is the substrate-binding site. The structures identify residues that may have important roles in catalysis, which have been confirmed by our mutagenesis studies. PMID- 17135237 TI - An insight into the mechanism of human cysteine dioxygenase. Key roles of the thioether-bonded tyrosine-cysteine cofactor. AB - Cysteine dioxygenase is a non-heme mononuclear iron metalloenzyme that catalyzes the oxidation of cysteine to cysteine sulfinic acid with addition of molecular dioxygen. This irreversible oxidative catabolism of cysteine initiates several important metabolic pathways related to diverse sulfurate compounds. Cysteine dioxygenase is therefore very important for maintaining the proper hepatic concentration of intracellular free cysteine. Mechanisms for mouse and rat cysteine dioxygenases have recently been reported based on their crystal structures in the absence of substrates, although there is still a lack of direct evidence. Here we report the first crystal structure of human cysteine dioxygenase in complex with its substrate L-cysteine to 2.7A, together with enzymatic activity and metal content assays of several single point mutants. Our results provide an insight into a new mechanism of cysteine thiol dioxygenation catalyzed by cysteine dioxygenase, which is tightly associated with a thioether bonded tyrosine-cysteine cofactor involving Tyr-157 and Cys-93. This cross-linked protein-derived cofactor plays several key roles different from those in galactose oxidase. This report provides a new potential target for therapy of diseases related to human cysteine dioxygenase, including neurodegenerative and autoimmune diseases. PMID- 17135238 TI - Differential effects of endoplasmic reticulum stress-induced autophagy on cell survival. AB - Autophagy is a cellular response to adverse environment and stress, but its significance in cell survival is not always clear. Here we show that autophagy could be induced in the mammalian cells by chemicals, such as A23187, tunicamycin, thapsigargin, and brefeldin A, that cause endoplasmic reticulum stress. Endoplasmic reticulum stress-induced autophagy is important for clearing polyubiquitinated protein aggregates and for reducing cellular vacuolization in HCT116 colon cancer cells and DU145 prostate cancer cells, thus mitigating endoplasmic reticulum stress and protecting against cell death. In contrast, autophagy induced by the same chemicals does not confer protection in a normal human colon cell line and in the non-transformed murine embryonic fibroblasts but rather contributes to cell death. Thus the impact of autophagy on cell survival during endoplasmic reticulum stress is likely contingent on the status of cells, which could be explored for tumor-specific therapy. PMID- 17135239 TI - A structural perspective on the interaction between lipopolysaccharide and factor C, a receptor involved in recognition of Gram-negative bacteria. AB - The recognition of broadly conserved microorganism components known as pathogen associated molecular patterns is an essential step in initiating the innate immune response. In the horseshoe crab, stimulation of hemocytes with lipopolysaccharide (LPS) causes the activation of its innate immune response, and Factor C, a serine protease zymogen, plays an important role in this event. Here, we report that Factor C associates with LPS on the hemocyte surface and directly recognizes Gram-negative bacteria. Structure-function analyses reveal that the LPS binding site is present in the N-terminal cysteine-rich (Cys-rich) region of the molecule and that it contains a tripeptide sequence consisting of an aromatic residue flanked by two basic residues that is conserved in other mammalian LPS recognizing proteins. Moreover, we have demonstrated that the Cys-rich region specifically binds to LPS on Gram-negative bacteria and that mutations in the tripeptide motif abrogate its association with both LPS and Gram-negative bacteria, underscoring the importance of the tripeptide in LPS interaction. Although the innate immune response to LPS in the horseshoe crab is distinct from that of mammals, it appears to rely on structural features that are conserved among LPS-recognizing proteins from diverse species. PMID- 17135240 TI - Regulation of EphA2 receptor endocytosis by SHIP2 lipid phosphatase via phosphatidylinositol 3-Kinase-dependent Rac1 activation. AB - Endocytosis of Eph receptors is critical for a number of biological processes, including modulating axon growth cone collapse response and regulating cell surface levels of receptor in epithelial cells. In particular, ephrin-A ligand stimulation of tumor cells induces EphA2 receptor internalization and degradation, a process that has been explored as a means to reduce tumor malignancy. However, the mechanism and regulation of ligand-induced Eph receptor internalization are not well understood. Here we show that SHIP2 (Src homology 2 domain-containing phosphoinositide 5-phosphatase 2) is recruited to activated EphA2 via a heterotypic sterile alpha motif (SAM)-SAM domain interaction, leading to regulation of EphA2 internalization. Overexpression of SHIP2 inhibits EphA2 receptor endocytosis, whereas suppression of SHIP2 expression by small interfering RNA-mediated gene silencing promotes ligand-induced EphA2 internalization and degradation. SHIP2 regulates EphA2 endocytosis via phosphatidylinositol 3-kinase-dependent Rac1 activation. Phosphatidylinositol 3,4,5-trisphosphate levels are significantly elevated in SHIP2 knockdown cells, phosphatidylinositol 3-kinase inhibitor decreases phosphatidylinositol 3,4,5 trisphosphate levels and suppresses increased EphA2 endocytosis. Ephrin-A1 stimulation activates Rac1 GTPase, and the Rac1-GTP levels are further increased in SHIP2 knockdown cells. A dominant negative Rac1 GTPase effectively inhibited ephrin-A1-induced EphA2 endocytosis. Together, our findings provide evidence that recruitment of SHIP2 to EphA2 attenuates a positive signal to receptor endocytosis mediated by phosphatidylinositol 3-kinase and Rac1 GTPase. PMID- 17135241 TI - Structural and mechanistic studies on the inhibition of the hypoxia-inducible transcription factor hydroxylases by tricarboxylic acid cycle intermediates. AB - In humans both the levels and activity of the alpha-subunit of the hypoxia inducible transcription factor (HIF-alpha) are regulated by its post-translation hydroxylation as catalyzed by iron- and 2-oxoglutarate (2OG)-dependent prolyl and asparaginyl hydroxylases (PHD1-3 and factor-inhibiting HIF (FIH), respectively). One consequence of hypoxia is the accumulation of tricarboxylic acid cycle intermediates (TCAIs). In vitro assays were used to assess non-2OG TCAIs as inhibitors of purified PHD2 and FIH. Under the assay conditions, no significant FIH inhibition was observed by the TCAIs or pyruvate, but fumarate, succinate, and isocitrate inhibited PHD2. Mass spectrometric analyses under nondenaturing conditions were used to investigate the binding of TCAIs to PHD2 and supported the solution studies. X-ray crystal structures of FIH in complex with Fe(II) and fumarate or succinate revealed similar binding modes for each in the 2OG co substrate binding site. The in vitro results suggest that the cellular inhibition of PHD2, but probably not FIH, by fumarate and succinate may play a role in the Warburg effect providing that appropriate relative concentrations of the components are achieved under physiological conditions. PMID- 17135242 TI - Dissecting the senescence-like program in tumor cells activated by Ras signaling. AB - Activated Ras signaling can induce a permanent growth arrest in osteosarcoma cells. Here, we report that a senescence-like growth inhibition is also achieved in human carcinoma cells upon the transduction of H-Ras(V12). Ras-induced tumor senescence can be recapitulated by the transduction of activated, but not wild type, MEK. The ability for H-Ras(V12) to suppress tumor cell growth is drastically compromised in cells that harbor endogenous activating ras mutations. Notably, growth inhibition of tumor cells containing ras mutations can be achieved through the introduction of activated MEK. Tumor senescence induced by Ras signaling can occur in the absence of p16 or Rb and is not interrupted by the inactivation of Rb, p107, or p130 via short hairpin RNA or the transduction with HPV16 E7. In contrast, inactivation of p21 via short hairpin RNA disrupts Ras induced tumor senescence. In summary, this study uncovers a senescence-like program activated by Ras signaling to inhibit cancer cell growth. This program appears to be intact in cancer cells that do not harbor ras mutations. Moreover, cancer cells that carry ras mutations remain susceptible to tumor senescence induced by activated MEK. These novel findings can potentially lead to the development of innovative cancer intervention. PMID- 17135243 TI - Abrogation of the transactivation activity of p53 by BCCIP down-regulation. AB - The tumor suppression function of p53 is mostly conferred by its transactivation activity, which is inactivated by p53 mutations in approximately 50% of human cancers. In cancers harboring wild type p53, the p53 transactivation activity may be compromised by other mechanisms. Identifying the mechanisms by which wild type p53 transactivation activity can be abrogated may provide insights into the molecular etiology of cancers harboring wild type p53. In this report, we show that BCCIP, a BRCA2 and CDKN1A-interacting protein, is required for the transactivation activity of wild type p53. In p53 wild type cells, BCCIP knock down by RNA interference diminishes the transactivation activity of p53 without reducing the p53 protein level, inhibits the binding of p53 to the promoters of p53 target genes p21 and HDM2, and reduces the tetrameric formation of p53. These data demonstrate a critical role of BCCIP in maintaining the transactivation activity of wild type p53 and further suggest down-regulation of BCCIP as a novel mechanism to impair the p53 function in cells harboring wild type p53. PMID- 17135244 TI - P2X7 nucleotide receptors mediate blebbing in osteoblasts through a pathway involving lysophosphatidic acid. AB - Extracellular nucleotides, released in response to mechanical or inflammatory stimuli, signal through P2 receptors in many cell types, including osteoblasts. P2X7 receptors are ATP-gated cation channels that can induce formation of large membrane pores. Disruption of the gene encoding the P2X7 receptor leads to decreased periosteal bone formation and insensitivity of the skeleton to mechanical stimulation. Our purpose was to investigate signaling pathways coupled to P2X7 activation in osteoblasts. Live cell imaging showed that ATP or 2 ',3 '-O (4-benzoylbenzoyl)-ATP (BzATP), but not UTP, UDP, or 2-methylthio-ADP, induced dynamic membrane blebbing in calvarial osteoblasts. Blebbing was observed in calvarial cells from wildtype but not P2X7 knock-out mice. P2X7 receptors coupled to activation of phospholipase D and A2, inhibition of which suppressed BzATP induced blebbing. Activation of these phospholipases leads to production of lysophosphatidic acid (LPA). LPA caused dynamic blebbing in osteoblasts from both wild-type and P2X7 knock-out mice, similar to that induced by BzATP in wildtype cells. However, LPA-induced blebbing was more rapid in onset and was not affected by inhibition of phospholipase D or A2. Blockade or desensitization of LPA receptors suppressed blebbing in response to LPA and BzATP, without affecting P2X7-stimulated pore formation. Thus, LPA functions downstream of P2X7 receptors to induce membrane blebbing. Furthermore, inhibition of Rho-associated kinase abolished blebbing induced by both BzATP and LPA. In summary, we propose a novel signaling axis that links P2X7 receptors through phospholipases to production of LPA and activation of Rho-associated kinase. This pathway may contribute to P2X7 stimulated osteogenesis during skeletal development and mechanotransduction. PMID- 17135245 TI - Functions of the multifaceted family of sphingosine kinases and some close relatives. PMID- 17135246 TI - Arachidonate-derived dihomoprostaglandin production observed in endotoxin stimulated macrophage-like cells. AB - Eicosanoids, including the prostaglandins, leukotrienes, hydroxyeicosatetraenoic acids, epoxyeicosatetraenoic acids, and related compounds, are biosynthetic, bioactive mediators derived from arachidonic acid (AA), a 20:4(n-6) fatty acid. We have developed a comprehensive and sensitive mass spectral analysis to survey eicosanoid release from endotoxin-stimulated RAW 264.7 macrophage-like cells that is capable of detecting over 70 diverse eicosanoids and eicosanoid metabolites, should they be present. We now address the question: Are biologically significant eicosanoids being overlooked? Herein, we illustrate a general approach to diverse isotope metabolic profiling of labeled exogenous substrates using mass spectrometry (DIMPLES/MS), demonstrated for one substrate (AA) and its resultant products (eicosanoids). RAW cells were incubated in medium supplemented with deuterium-labeled AA. When the cells are stimulated, two sets of eicosanoids are produced, one from endogenous AA and the other from the supplemented (exogenous) deuterium-labeled form. This produces a signature mass spectral "doublet" pattern, allowing for a comprehensive and diverse eicosanoid search requiring no previous knowledge or assumptions as to what these species may be, in contrast to traditional methods. We report herein observing unexpected AA metabolites generated by the cells, some of which may constitute novel bioactive eicosanoids or eicosanoid inactivation metabolites, as well as demonstrating differing metabolic pathways for the generation of isomeric prostaglandins and potential peroxisome proliferator-activated receptor activators. Unexpectedly, we report observing a series of 1a, 1b-dihomologue prostaglandins, products of adrenic acid (22:4(n-6)), resulting from the two-carbon elongation of AA by the RAW cells. PMID- 17135247 TI - Staphylococcus aureus subvert autophagy for induction of caspase-independent host cell death. AB - Staphylococcus aureus is a common bacterial etiology of serious infectious diseases. S. aureus can invade various types of non-professional phagocytes to produce host cell death. We show here that shortly after invasion of HeLa cells S. aureus transit to autophagosomes was characterized by double membranes and co localization with LC3. S. aureus were not able to replicate and produce cell death in autophagy-deficient atg5-/- mouse embryonic fibroblasts. S. aureus containing autophagosomes do not acidify nor do they acquire lysosome-associated membrane protein-2, indicating that S. aureus inhibits autophagosome maturation and fusion with lysosomes. Eventually, S. aureus escape from autophagosomes into the cytoplasm, which results in caspase-independent host cell death. S. aureus strains deficient for agr, a global regulator of S. aureus virulence, were not targeted by autophagy and did not produce host-cell death. Autophagy induction by rapamycin restored both replication and cytotoxicity of agr-deficient S. aureus strains, indicating that an agr-regulated factor(s) is required for autophagy mediated cytotoxicity. The results of this study suggest that rapid induction of autophagy is essential for S. aureus replication, escape into the cytoplasm, and host cell killing. PMID- 17135248 TI - Polo-like kinases inhibited by wortmannin. Labeling site and downstream effects. AB - Polo-like kinases play crucial roles throughout mitosis. We previously reported that wortmannin potently inhibits Polo-like kinase 1 (Plk1). In this study, we show that wortmannin also strongly inhibits Polo-like kinase 3 (Plk3). To further characterize this inhibition, we identified the sites of labeling on Plk1 and Plk3 targeted by AX7503, a tetramethylrhodamine-wortmannin conjugate. AX7503 labeling on Plk1 and Plk3 was found to occur on a conserved ATP binding site residue. In addition, we show that wortmannin inhibits Plk3 activity in live cells at concentrations commonly used to inhibit the more well known targets of wortmannin, the phosphoinositide 3-kinases. Importantly, we found that inhibition of Plk3 by wortmannin lead to a decrease in phosphorylation of p53 on serine 20 induced by DNA damage, demonstrating the effect of wortmannin on a downstream Plk3 target. Taken together, our results suggest that wortmannin can affect multiple functions of Plk3 in cell cycle progression and at the DNA damage check point. The identification of the labeling sites of Plk1 and Plk3 by AX7503 may be useful in designing more effective compounds to target Polo-like kinases for cancer treatment and also may be useful for the structural study of Plk domains. PMID- 17135249 TI - An E2F/miR-20a autoregulatory feedback loop. AB - The E2F family of transcription factors is essential in the regulation of the cell cycle and apoptosis. While the activity of E2F1-3 is tightly controlled by the retinoblastoma family of proteins, the expression of these factors is also regulated at the level of transcription, post-translational modifications and protein stability. Recently, a new level of regulation of E2Fs has been identified, where micro-RNAs (miRNAs) from the mir-17-92 cluster influence the translation of the E2F1 mRNA. We now report that miR-20a, a member of the mir-17 92 cluster, modulates the translation of the E2F2 and E2F3 mRNAs via binding sites in their 3'-untranslated region. We also found that the endogenous E2F1, E2F2, and E2F3 directly bind the promoter of the mir-17-92 cluster activating its transcription, suggesting an autoregulatory feedback loop between E2F factors and miRNAs from the mir-17-92 cluster. Our data also point toward an anti-apoptotic role for miR-20a, since overexpression of this miRNA decreased apoptosis in a prostate cancer cell line, while inhibition of miR-20a by an antisense oligonucleotide resulted in increased cell death after doxorubicin treatment. This anti-apoptotic role of miR-20a may explain some of the oncogenic capacities of the mir-17-92 cluster. Altogether, these results suggest that the autoregulation between E2F1-3 and miR-20a is important for preventing an abnormal accumulation of E2F1-3 and may play a role in the regulation of cellular proliferation and apoptosis. PMID- 17135250 TI - Protoporphyrin IX interacts with wild-type p53 protein in vitro and induces cell death of human colon cancer cells in a p53-dependent and -independent manner. AB - Photodynamic therapy (PDT) of cancer is an alternative treatment for tumors resistant to chemo- and radiotherapy. It induces cancer cell death mainly through generation of reactive oxygen species by a laser light-activated photosensitizer. It has been suggested that the p53 tumor suppressor protein sensitizes some human cancer cells to PDT. However, there is still no direct evidence for this. We have demonstrated here for the first time that the photosensitizer protoporphyrin IX (PpIX) binds to p53 and disrupts the interaction between p53 tumor suppressor protein and its negative regulator HDM2 in vitro and in cells. Moreover, HCT116 colon cancer cells exhibited a p53-dependent sensitivity to PpIX in a dose dependent manner, as was demonstrated using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay and fluorescence-activated cell sorter (FACS) analysis of cell cycle profiles. We have also observed induction of p53 target pro-apoptotic genes, e.g. puma (p53-up-regulated modulator of apoptosis), and bak in PpIX-treated cells. In addition, p53-independent growth suppression by PpIX was detected in p53-negative cells. PDT treatment (2 J/cm2) of HCT116 cells induced p53-dependent activation of pro-apoptotic gene expression followed by growth suppression and induction of apoptosis. PMID- 17135251 TI - Cochlear function in mice lacking the BK channel alpha, beta1, or beta4 subunits. AB - Large conductance voltage- and calcium-activated potassium (BK) channels are important for regulating many essential cellular functions, from neuronal action potential shape and firing rate to smooth muscle contractility. In amphibians, reptiles, and birds, BK channels mediate the intrinsic frequency tuning of the cochlear hair cell by an electrical resonance mechanism. In contrast, inner hair cells of the mammalian cochlea are extrinsically tuned by accessory structures of the cochlea. Nevertheless, BK channels are present in inner hair cells and encode a fast activating outward current. To understand the role of the BK channel alpha and beta subunits in mammalian inner hair cells, we analyzed the morphology, physiology, and function of these cells from mice lacking the BK channel alpha (Slo-/-) and also the beta1 and beta4 subunits (beta1/4-/-). Beta1/4-/- mice showed normal subcellular localization, developmental acquisition, and expression of BK channels. Beta1/4-/- mice showed normal cochlear function as indicated by normal auditory brainstem responses and distortion product otoacoustic emissions. Slo-/- mice also showed normal cochlear function despite the absence of the BKalpha subunit and the absence of fast activating outward current from the inner hair cells. Moreover, microarray analyses revealed no compensatory changes in transcripts encoding ion channels or transporters in the cochlea from Slo-/- mice. Slo-/- mice did, however, show increased resistance to noise-induced hearing loss. These findings reveal the fundamentally different contribution of BK channels to nonmammalian and mammalian hearing and suggest that BK channels should be considered a target in the prevention of noise-induced hearing loss. PMID- 17135252 TI - The VP16 activation domain establishes an active mediator lacking CDK8 in vivo. AB - VP16 has been widely used to unravel the mechanisms underlying gene transcription. Much of the previous work has been conducted in reconstituted in vitro systems. Here we study the formation of transcription complexes at stable reporters under the control of an inducible Tet-VP16 activator in living cells. In this simplified model for gene activation VP16 recruits the general factors and the cofactors Mediator, GCN5, CBP, and PC4, within minutes to the promoter region. Activation is accompanied by only minor changes in histone acetylation and H3K4 methylation but induces a marked promoter-specific increase in H3K79 methylation. Mediated through contacts with VP16 several subunits of the cleavage and polyadenylation factor (CPSF/CstF) are concentrated at the promoter region. We provide in vitro and in vivo evidence that VP16 activates transcription through a specific MED25-associated Mediator, which is deficient in CDK8. PMID- 17135253 TI - Compensatory mechanism for homeostatic blood pressure regulation in Ephx2 gene disrupted mice. AB - Arachidonic acid-derived epoxides, epoxyeicosatrienoic acids, are important regulators of vascular homeostasis and inflammation, and therefore manipulation of their levels is a potentially useful pharmacological strategy. Soluble epoxide hydrolase converts epoxyeicosatrienoic acids to their corresponding diols, dihydroxyeicosatrienoic acids, modifying or eliminating the function of these oxylipins. To better understand the phenotypic impact of Ephx2 disruption, two independently derived colonies of soluble epoxide hydrolase-null mice were compared. We examined this genotype evaluating protein expression, biofluid oxylipin profile, tissue oxylipin production capacity, and blood pressure. Ephx2 gene disruption eliminated soluble epoxide hydrolase protein expression and activity in liver, kidney, and heart from each colony. Plasma levels of epoxy fatty acids were increased, and fatty acid diols levels were decreased, while measured levels of lipoxygenase- and cyclooxygenase-dependent oxylipins were unchanged. Liver and kidney homogenates also show elevated epoxide fatty acids. However, in whole kidney homogenate a 4-fold increase in the formation of 20 hydroxyeicosatetraenoic acid was measured along with a 3-fold increase in lipoxygenase-derived hydroxylation and prostanoid production. Unlike previous reports, however, neither Ephx2-null colony showed alterations in basal blood pressure. Finally, the soluble epoxide hydrolase-null mice show a survival advantage following acute systemic inflammation. The data suggest that blood pressure homeostasis may be achieved by increasing production of the vasoconstrictor, 20-hydroxyeicosatetraenoic acid in the kidney of the Ephx2-null mice. This shift in renal metabolism is likely a metabolic compensation for the loss of the soluble epoxide hydrolase gene. PMID- 17135254 TI - A comprehensive structure-function map of the intracellular surface of the human C5a receptor. I. Identification of critical residues. AB - G protein-coupled receptors are one of the largest protein families in nature; however, the mechanisms by which they activate G proteins are still poorly understood. To identify residues on the intracellular face of the human C5a receptor that are involved in G protein activation, we performed a genetic analysis of each of the three intracellular loops and the carboxyl-terminal tail of the receptor. Amino acid substitutions were randomly incorporated into each loop, and functional receptors were identified in yeast. The third intracellular loop contains the largest number of preserved residues (positions resistant to amino acid substitutions), followed by the second loop, the first loop, and lastly the carboxyl terminus. Surprisingly, complete removal of the carboxyl terminal tail did not impair C5a receptor signaling. When mapped onto a three dimensional structural model of the inactive state of the C5a receptor, the preserved residues reside on one half of the intracellular surface of the receptor, creating a potential activation face. Together these data provide one of the most comprehensive functional maps of the intracellular surface of any G protein-coupled receptor to date. PMID- 17135255 TI - In vitro fluorescence anisotropy analysis of the interaction of full-length SRC1a with estrogen receptors alpha and beta supports an active displacement model for coregulator utilization. AB - Binding of full-length P160 coactivators to hormone response element-steroid receptor complexes has been difficult to investigate in vitro. Here, we report a new application of our recently described fluorescence anisotropy microplate assay to investigate binding and dissociation of full-length steroid receptor coactivator-1a (SRC1a) from full-length estrogen receptor alpha (ERalpha) or estrogen receptor beta (ERbeta) bound to a fluorescein-labeled (fl) estrogen response element (ERE). SRC1a exhibited slightly higher affinity binding to flERE.ERbeta than to flERE.ERalpha. Binding of SRC1a to flERE.ERalpha and to flERE.ERbeta was 17beta-estradiol (E2)-dependent and was nearly absent when ICI 182,780, raloxifene, or 4-hydroxytamoxifen were bound to the ERs. SRC1a binds to flERE.E2-ERalpha and flERE.E2-ERbeta complexes with a t1/2 of 15-20 s. Short LXXLL-containing nuclear receptor (NR) box peptides from P160 coactivators competed much better for SRC1a binding to flERE.E2-ER than an NR box peptide from TRAP220. However, approximately 40-250-fold molar excess of the P160 NR box peptides was required to inhibit SRC1a binding by 50%. This suggests that whereas the NR box region is a primary site of interaction between SRC1a and ERE.E2-ER, additional contacts between the coactivator and the ligand-receptor-DNA complex make substantial contributions to overall affinity. Increasing amounts of NR box peptides greatly enhanced the rate of dissociation of SRC1a from preformed flERE.E2-ER complexes. The data support a model in which coactivator exchange is facilitated by active displacement and is not simply the result of passive dissociation and replacement. It also shows that an isolated coactivator exhibits an inherent capacity for rapid exchange. PMID- 17135256 TI - Selectin ligand expression regulates the initial vascular interactions of colon carcinoma cells: the roles of CD44v and alternative sialofucosylated selectin ligands. AB - Selectin-mediated binding of tumor cells to platelets, leukocytes, and vascular endothelium may regulate their hematogenous spread in the microvasculature. We recently reported that CD44 variant isoforms (CD44v) on LS174T colon carcinoma cells possess selectin binding activity. Here we extended those findings by showing that T84 and Colo205 colon carcinoma cells bind selectins via sialidase sensitive O-linked glycans presented on CD44v, independent of heparan and chondroitin sulfate. To assess the functional role of CD44v in selectin-mediated binding, we quantified the adhesion to selectins of T84 cell subpopulations sorted based on their CD44 expression levels and stable LS174T cell lines generated using CD44 short hairpin RNA. High versus low CD44-expressing T84 cells tethered more efficiently to P- and L-selectin, but not E-selectin, and rolled more slowly on P- and E-selectin. Knocking down CD44 expression on LS174T cells inhibited binding to P-selectin and increased rolling velocities over P- and L selectin relative to control-transfected cells, without affecting tethering and rolling on E-selectin, however. Blot rolling analysis revealed the presence of alternative sialylated glycoproteins with molecular masses of approximately 170 and approximately 130 kDa, which can mediate selectin binding in CD44-knockdown cells. Heparin diminishes the avidity of colon carcinoma cells for P- and L selectin, which may compromise integrin-mediated firm adhesion to host cells and mitigate metastasis. Our finding that CD44v is a functional P-selectin ligand on colon carcinoma provides a novel perspective on the enhanced metastatic potential associated with tumor CD44v overexpression and the role of selectins in metastasis. PMID- 17135257 TI - Monoclonal antibody clearance. Impact of modulating the interaction of IgG with the neonatal Fc receptor. AB - The neonatal Fc receptor (FcRn) plays a critical role in regulating IgG homeostasis in vivo. There are mixed reports on whether modification of the interaction with FcRn can be used as an engineering strategy to improve the pharmacokinetic and pharmacodynamic properties of monoclonal antibodies. We tested whether the T250Q/M428L mutations, which improved the pharmacokinetics of humanized IgGs in the rhesus monkey, would translate to a pharmacokinetic benefit in both cynomolgus monkeys and mice when constructed on a different humanized IgG framework (anti-tumor necrosis factor-alpha (TNFalpha)). The T250Q/M428L anti TNFalpha variant displayed an approximately 40-fold increase in binding affinity to cynomolgus monkey FcRn (C-FcRn) at pH 6.0, with maintenance of the pH binding dependence. We also constructed another anti-TNFalpha variant (P257I/Q311I) whose binding kinetics with the C-FcRn was similar to that of the T250Q/M428L variant. The binding affinity of the T250Q/M428L variant for murine FcRn was increased approximately 500-fold, with maintenance of pH dependence. In contrast to the interaction with C-FcRn, this interaction was driven mainly by a decrease in the rate of dissociation. Despite the improved in vitro binding properties of the anti-TNFalpha T250Q/M428L and P257I/Q311I variants to C-FcRn, the pharmacokinetic profiles of these molecules were not differentiated from the wild-type antibody in cynomolgus monkeys after intravenous administration. When administered intravenously to mice, the T250Q/M428L anti-TNFalpha variant displayed improved pharmacokinetics, characterized by an approximately 2-fold slower clearance than the wild-type antibody. The discrepancy between these data and previously reported benefits in rhesus monkeys and the inability of these mutations to translate to improved kinetics across species may be related to a number of factors. We propose extending consideration to differences in the absolute IgG FcRn affinity, the kinetics of the IgG/FcRn interaction, and differences in the relative involvement of this pathway in the context of other factors influencing the disposition or elimination of monoclonal antibodies. PMID- 17135258 TI - Regulation of calpain activity in rat brain with altered Ca2+ homeostasis. AB - Activation of calpain occurs as an early event in correlation with an increase in [Ca2+]i induced in rat brain upon treatment with a high salt diet for a prolonged period of time. The resulting sequential events have been monitored in the brain of normal and hypertensive rats of the Milan strain, diverging for a constitutive alteration in the level of [Ca2+]i found to be present in nerve cells of hypertensive animals. After 2 weeks of treatment, the levels of the plasma membrane Ca2+-ATPase and of native calpastatin are profoundly decreased. These degradative processes, more pronounced in the brain of hypertensive rats, are progressively and efficiently compensated in the brain of both rat strains by different incoming mechanisms. Along with calpastatin degradation, 15-kDa still active inhibitory fragments are accumulated, capable of efficiently replacing the loss of native inhibitor molecules. A partial return to a more efficient control of Ca2+ homeostasis occurs in parallel, assured by an early increase in the expression of Ca2+-ATPase and of calpastatin, both producing, after 12 weeks of a high salt (sodium) diet, the restoration of almost original levels of the Ca2+ pump and of significant amounts of native inhibitor molecules. Thus, conservative calpastatin fragmentation, associated with an increased expression of Ca2+-ATPase and of the calpain natural inhibitor, has been demonstrated to occur in vivo in rat brain. This represents a sequential adaptive response capable of overcoming the effects of calpain activation induced by a moderate long term elevation of [Ca2+]i. PMID- 17135259 TI - Resistance to Bacillus thuringiensis toxin in Caenorhabditis elegans from loss of fucose. AB - A mutation in the Caenorhabditis elegans bre-1 gene was isolated in a screen for Bacillus thuringiensis toxin-resistant (bre) mutants to the Cry5B crystal toxin made by B. thuringiensis. bre-1 mutant animals are different from the four other cloned bre mutants in that their level of resistance is noticeably lower. bre-1 animals also display a significantly reduced brood size at 25 degrees C. Here we cloned the bre-1 gene and characterized the bre-1 mutant phenotype. bre-1 encodes a protein with significant homology to a GDP-mannose 4,6-dehydratase, which catalyzes the first step in the biosynthesis of GDP-fucose from GDP-mannose. Injection of GDP-fucose but not fucose into C. elegans intestinal cells rescues bre-1 mutant phenotypes. Thus, C. elegans lacks a functional fucose salvage pathway. Furthermore, we demonstrate that bre-1 mutant animals are defective in production of fucosylated glycolipids and that bre-1 mutant animals make quantitatively reduced levels of glycolipid receptors for Cry5B. We finally show that bre-1 mutant animals, although viable, show a lack of fucosylated N- and O glycans, based on mass spectrometric evidence. Thus, C. elegans can survive with little fucose and can develop resistance to crystal toxin by loss of a monosaccharide biosynthetic pathway. PMID- 17135260 TI - Loss of assembly of the main basement membrane collagen, type IV, but not fibril forming collagens and embryonic death in collagen prolyl 4-hydroxylase I null mice. AB - Collagen prolyl 4-hydroxylases (C-P4Hs) catalyze the formation of the 4 hydroxyproline residues that are essential for the generation of triple helical collagen molecules. The vertebrate C-P4Hs I, II, and III are [alpha(I)]2beta2, [alpha(II)]2beta2, and [alpha(III)]2beta2 tetramers with identical beta subunits. We generated mice with targeted inactivation of the P4ha1 gene encoding the catalytic alpha subunit of C-P4H I to analyze its specific functions. The null mice died after E10.5, showing an overall developmental delay and a dilated endoplasmic reticulum in their cells. The capillary walls were frequently ruptured, but the capillary density remained unchanged. The C-P4H activity level in the null embryos and fibroblasts cultured from them was 20% of that in the wild type, being evidently due to the other two isoenzymes. Collagen IV immunofluorescence was almost absent in the basement membranes of the null embryos, and electron microscopy revealed disrupted basement membranes, while immunoelectron microscopy showed a lack of collagen IV in them. The amount of soluble collagen IV was increased in the null embryos and cultured null fibroblasts, indicating a lack of assembly of collagen IV molecules into insoluble structures, probably due to their underhydroxylation and hence abnormal conformation. In contrast, the null embryos had collagen I and III fibrils with a typical cross-striation pattern but slightly increased diameters, and the null fibroblasts secreted fibril-forming collagens, although less efficiently than wild-type cells. The primary cause of death of the null embryos was thus most likely an abnormal assembly of collagen IV. PMID- 17135261 TI - Airway epithelial cell migration and wound repair by ATP-mediated activation of dual oxidase 1. AB - The airway epithelium is continuously subjected to environmental pollutants, airborne pathogens, and allergens and relies on several intrinsic mechanisms to maintain barrier integrity and to promote epithelial repair processes following injury. Here, we report a critical role for dual oxidase 1 (Duox1), a newly identified NADPH oxidase homolog within the tracheobronchial epithelium, in airway epithelial cell migration and repair following injury. Activation of Duox1 during epithelial injury is mediated by cellular release of ATP, which signals through purinergic receptors expressed on the epithelial cell surface. Purinergic receptor stimulation by extracellular ATP is a critical determinant of epithelial cell migration and repair following injury and is associated with activation of extracellular signal-regulated kinases (ERK1/2) and matrix metalloproteinase-9 (MMP-9). Stimulation of these integral features of epithelial cell migration and repair processes was found to require the activation of Duox1. Our findings demonstrate a novel role for Duox1 in the tracheobronchial epithelium, in addition to its proposed role in antimicrobial host defense, by participating in epithelial repair processes to maintain epithelial integrity and barrier function in the face of environmental stress. PMID- 17135262 TI - A ganglioside-induced toxic soluble Abeta assembly. Its enhanced formation from Abeta bearing the Arctic mutation. AB - The mechanism underlying plaque-independent neuronal death in Alzheimer disease (AD), which is probably responsible for early cognitive decline in AD patients, remains unclarified. Here, we show that a toxic soluble Abeta assembly (TAbeta) is formed in the presence of liposomes containing GM1 ganglioside more rapidly and to a greater extent from a hereditary variant-type ("Arctic") Abeta than from wild-type Abeta. TAbeta is also formed from soluble Abeta through incubation with natural neuronal membranes prepared from aged mouse brains in a GM1 ganglioside dependent manner. An oligomer-specific antibody (anti-Oligo) significantly suppresses TAbeta toxicity. Biophysical and structural analyses by atomic force microscopy and size exclusion chromatography revealed that TAbeta is spherical with diameters of 10-20 nm and molecular masses of 200-300 kDa. TAbeta induces neuronal death, which is abrogated by the small interfering RNA-mediated knockdown of nerve growth factor receptors, including TrkA and p75 neurotrophin receptor. Our results suggest that soluble Abeta assemblies, such as TAbeta, can cause plaque-independent neuronal death that favorably occurs in nerve growth factor-dependent neurons in the cholinergic basal forebrain in AD. PMID- 17135263 TI - A role for protein misfolding in immunogenicity of biopharmaceuticals. AB - For largely unknown reasons, biopharmaceuticals evoke potentially harmful antibody formation. Such antibodies can inhibit drug efficacy and, when directed against endogenous proteins, cause life-threatening complications. Insight into the mechanisms by which biopharmaceuticals break tolerance and induce an immune response will contribute to finding solutions to prevent this adverse effect. Using a transgenic mouse model, we here demonstrate that protein misfolding, detected with the use of tissue-type plasminogen activator and thioflavin T, markers of amyloid-like properties, results in breaking of tolerance. In wild type mice, misfolding enhances protein immunogenicity. Several commercially available biopharmaceutical products were found to contain misfolded proteins. In some cases, the level of misfolded protein was found to increase upon storage under conditions prescribed by the manufacturer. Our results indicate that misfolding of therapeutic proteins is an immunogenic signal and a risk factor for immunogenicity. These findings offer novel possibilities to detect immunogenic protein entities with tPA and reduce immunogenicity of biopharmaceuticals. PMID- 17135264 TI - Nitric oxide mediates natural polyphenol-induced Bcl-2 down-regulation and activation of cell death in metastatic B16 melanoma. AB - Intravenous administration to mice of trans-pterostilbene (t-PTER; 3,5-dimethoxy 4'-hydroxystilbene) and quercetin (QUER; 3,3',4',5,6-pentahydroxyflavone), two structurally related and naturally occurring small polyphenols, inhibits metastatic growth of highly malignant B16 melanoma F10 (B16M-F10) cells. t-PTER and QUER inhibit bcl-2 expression in metastatic cells, which sensitizes them to vascular endothelium-induced cytotoxicity. However, the molecular mechanism(s) linking polyphenol signaling and bcl-2 expression are unknown. NO is a potential bioregulator of apoptosis with controversial effects on Bcl-2 regulation. Polyphenols may affect NO generation. Short-term exposure (60 min/day) to t-PTER (40 microM) and QUER (20 microM) (approximate mean values of the plasma concentrations measured within the first hour after intravenous administration of 20 mg of each polyphenol/kg) down-regulated inducible NO synthetase in B16M-F10 cells and up-regulated endothelial NO synthetase in the vascular endothelium and thereby facilitated endothelium-induced tumor cytotoxicity. Very low and high NO levels down-regulated bcl-2 expression in B16M-F10 cells. t-PTER and QUER induced a NO shortage-dependent decrease in cAMP-response element-binding protein phosphorylation, a positive regulator of bcl-2 expression, in B16M-F10 cells. On the other hand, during cancer and endothelial cell interaction, t-PTER- and QUER induced NO release from the vascular endothelium up-regulated neutral sphingomyelinase activity and ceramide generation in B16M-F10 cells. Direct NO induced cytotoxicity and ceramide-induced mitochondrial permeability transition and apoptosis activation can explain the increased endothelium-induced death of Bcl-2-depleted B16M-F10 cells. PMID- 17135265 TI - Human Ikaros function in activated T cells is regulated by coordinated expression of its largest isoforms. AB - The Ikaros gene is alternately spliced to generate multiple zinc finger proteins involved in gene regulation and chromatin remodeling. Whereas murine studies have provided important information regarding the role of Ikaros in the mouse, little is known of Ikaros function in human. We report functional analyses of the two largest human Ikaros (hIK) isoforms, hIK-VI and hIK-H, in T cells. Abundant expression of hIK-H, the largest described isoform, is restricted to human hematopoietic cells. We find that the DNA binding affinity of hIK-H differs from that of hIK-VI. Co-expression of hIk-H with hIk-VI alters the ability of Ikaros complexes to bind DNA motifs found in pericentromeric heterochromatin (PC-HC). In the nucleus, hIK-VI is localized solely in PC-HC, whereas the hIK-H protein exhibits dual centromeric and non-centromeric localization. Mutational analysis defined the amino acids responsible for the distinct DNA binding ability of hIK H, as well as the sequence required for the specific subcellular localization of this isoform. In proliferating cells, the binding of hIK-H to the upstream regulatory region of known Ikaros target genes correlates with their positive regulation by Ikaros. Results suggest that expression of hIK-H protein restricts affinity of Ikaros protein complexes toward specific PC-HC repeats. We propose a model, whereby the binding of hIK-H-deficient Ikaros complexes to the regulatory sequence of target genes would recruit these genes to the restrictive pericentromeric compartment, resulting in their repression. The presence of hIK-H in the Ikaros complex would alter its affinity for PC-HC, leading to chromatin remodeling and activation of target genes. PMID- 17135266 TI - Functional and structural aspects of poplar cytosolic and plastidial type a methionine sulfoxide reductases. AB - The genome of Populus trichocarpa contains five methionine sulfoxide reductase A genes. Here, both cytosolic (cMsrA) and plastidial (pMsrA) poplar MsrAs were analyzed. The two recombinant enzymes are active in the reduction of methionine sulfoxide with either dithiothreitol or poplar thioredoxin as a reductant. In both enzymes, five cysteines, at positions 46, 81, 100, 196, and 202, are conserved. Biochemical and enzymatic analyses of the cysteine-mutated MsrAs support a catalytic mechanism involving three cysteines at positions 46, 196, and 202. Cys(46) is the catalytic cysteine, and the two C-terminal cysteines, Cys(196) and Cys(202), are implicated in the thioredoxin-dependent recycling mechanism. Inspection of the pMsrA x-ray three-dimensional structure, which has been determined in this study, strongly suggests that contrary to bacterial and Bos taurus MsrAs, which also contain three essential Cys, the last C-terminal Cys(202), but not Cys(196), is the first recycling cysteine that forms a disulfide bond with the catalytic Cys(46). Then Cys(202) forms a disulfide bond with the second recycling cysteine Cys(196) that is preferentially reduced by thioredoxin. In agreement with this assumption, Cys(202) is located closer to Cys(46) compared with Cys(196) and is included in a (202)CYG(204) signature specific for most plant MsrAs. The tyrosine residue corresponds to the one described to be involved in substrate binding in bacterial and B. taurus MsrAs. In these MsrAs, the tyrosine residue belongs to a similar signature as found in plant MsrAs but with the first C-terminal cysteine instead of the last C-terminal cysteine. PMID- 17135267 TI - SCLIP, a microtubule-destabilizing factor, interacts with RasGRF1 and inhibits its ability to promote Rac activation and neurite outgrowth. AB - RasGRF1 is a neuron-specific guanine nucleotide exchange factor for the small GTPases Ras and Rac. It is implicated in the regulation of memory formation and in the development of tolerance to drug abuse, although the mechanisms have been elucidated only in part. Here we report the isolation, by the yeast two-hybrid screen, of the microtubule-destabilizing factor SCLIP (SCG10-like protein) as a novel RasGRF1-interacting protein. This interaction requires the region spanning the Dbl-homology domain of RasGRF1, endowed with catalytic activity on Rac. In search for a possible function we found by biochemical means that SCLIP influences the signaling properties of RasGRF1, greatly reducing its ability to activate the Rac/p38 MAPK pathway, while the Ras/Erk one remains unaffected. Moreover, a potential role is suggested by transfection studies in neuronal PC12 cells in which RasGRF1 induces neurite outgrowth, and coexpression of SCLIP counteracts this effect, causing a dramatic decrease in the percentage of cells bearing neurites, which also appear significantly shortened. This study unveils a physical and functional interaction between RasGRF1 and SCLIP. We suggest that this novel interplay may have possible implications in mechanisms that regulate neuronal morphology and structural plasticity. PMID- 17135268 TI - Direct regulation of an oncogenic micro-RNA cluster by E2F transcription factors. AB - Micro-RNAs (miRNAs) are a class of non-coding RNAs that post-transcriptionally regulate gene expression via the RNA interference pathway. In addition to roles in normal development, miRNAs have recently been implicated in a range of human diseases, including cancer. We recently demonstrated that a polycistronic cluster of miRNAs, miR-17-92, is oncogenic in a mouse model for Burkitt's lymphoma. This is due, in part, to a reduced apoptotic program. In an effort to understand the regulation of miR-17-92, we have studied the promoter structure of this miRNA cluster. The primary transcript initiates from a consensus initiator sequence downstream of a nonconsensus TATA box. The core promoter region contains two functional E2F transcription factor binding sites. Chromatin immunoprecipitation demonstrates that E2F3 is the primary E2F family member that occupies the promoter. These data place miR-17-92 in a regulatory loop between E2F3 and the miR-17 target E2F1. We propose a model whereby miR-17-92 promotes cell proliferation by shifting the E2F transcriptional balance away from the pro apoptotic E2F1 and toward the proliferative E2F3 transcriptional network. PMID- 17135269 TI - Fas-activated serine/threonine kinase (FAST K) synergizes with TIA-1/TIAR proteins to regulate Fas alternative splicing. AB - The factors and mechanisms that mediate the effects of intracellular signaling cascades on alternative pre-mRNA splicing are poorly understood. TIA-1 (T-cell intracellular antigen 1) and TIAR (TIA-1-related) proteins regulate alternative pre-mRNA splicing by promoting the use of suboptimal 5' splice sites followed by uridine-rich intronic enhancer sequences. These proteins promote, for example, inclusion of Fas receptor exon 6, which leads to an mRNA encoding a pro-apoptotic form of the receptor at the expense of the form that skips exon 6, which encodes an anti-apoptotic form. Fas-activated serine/threonine kinase (FAST K) is known to interact with and phosphorylate TIA-1. Here we have tested the possibility that FAST K influences alternative pre-mRNA splicing by affecting the activity of TIA-1/TIAR. Depletion of FAST K form Jurkat cells leads to skipping of exon 6 from endogenous Fas transcripts. Conversely, FAST K overexpression enhances exon 6 inclusion of Fas reporters transfected in HeLa cells. Consistent with the possibility that the effects of FAST K are mediated by changes in the function of TIA-1/TIAR, the effects of FAST K overexpression (i) are largely suppressed by depletion of TIA-1 and TIAR and (ii) are significantly compromised by mutation of a TIA-1/TIAR-responsive enhancer present downstream of exon 6 5' splice site. Furthermore, in vitro phosphorylation of TIA-1 by FAST K results in enhanced U1 snRNP recruitment. Interestingly, this enhancement is not due to increased binding of TIA-1 to the pre-mRNA. Taken together, the results connect Fas signaling with the activity of splicing factors that modulate Fas alternative splicing, suggesting the existence of an autoregulatory loop that could serve to amplify Fas responses. PMID- 17135270 TI - The role of the C-terminal domain of protein tyrosine phosphatase-1B in phosphatase activity and substrate binding. AB - Protein tyrosine phosphatase 1B (PTP-1B) has been implicated in the regulation of the insulin receptor. Dephosphorylation of the insulin receptor results in decreased insulin signaling and thus decreased glucose uptake. PTP-1B-/- mice have increased insulin sensitivity and are resistant to weight gain when fed a high fat diet, validating PTP-1B as a potential target for the treatment of type 2 diabetes. Many groups throughout the world have been searching for selective inhibitors for PTP-1B, and most of them target inhibitors to PTP-1B-(1-298), the N-terminal catalytic domain of the enzyme. However, the C-terminal domain is quite large and could influence the activity of the enzyme. Using two constructs of PTP-1B and a phosphopeptide as substrate, steady state assays showed that the presence of the C-terminal domain decreased both the Km and the k(cat) 2-fold. Pre-steady state kinetic experiments showed that the presence of the C-terminal domain improved the affinity of the enzyme for a phosphopeptide 2-fold, primarily because the off-rate was slower. This suggests that the C-terminal domain of PTP 1B may contact the phosphopeptide in some manner, allowing it to remain at the active site longer. This could be useful when screening libraries of compounds for inhibitors of PTP-1B. A compound that is able to make contacts with the C terminal domain of PTP-1B would not only have a modest improvement in affinity but may also provide for specificity over other phosphatases. PMID- 17135271 TI - Site-specific Lys-63-linked tumor necrosis factor receptor-associated factor 6 auto-ubiquitination is a critical determinant of I kappa B kinase activation. AB - Tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6) is a key mediator in proximal signaling of the interleukin-1/Toll-like receptor and the TNF receptor superfamily. Analysis of TRAF6-deficient mice revealed a fundamental role of TRAF6 in osteoclastogenesis; however, the molecular mechanism underlying TRAF6 signaling in this biological process is not understood. Recent biochemical evidence has indicated that TRAF6 possesses ubiquitin ligase activity that controls the activation of IKK and NF-kappaB. Because these studies are primarily based on cell-free systems, the role of the ubiquitin ligase activity of TRAF6 and its auto-ubiquitination to initiate the NF-kappaB pathway in vivo remain elusive. Here we show that an intact RING domain of TRAF6 in conjunction with the E2 enzyme Ubc13/Uev1A is necessary for Lys-63-linked auto-ubiquitination of TRAF6 and for its ability to activate IKK and NF-kappaB. Furthermore, a RING mutant of TRAF6 abolishes its ability to induce receptor activator of NF-kappaB-independent osteoclast differentiation and nuclear accumulation of the transcription factor NFATc1. Notably, we map the auto-ubiquitination site of TRAF6 to a single Lys residue, which if mutated renders TRAF6 unable to activate transforming growth factor-beta-activated kinase 1 and IKK and to cause spontaneous osteoclast differentiation. Additionally, we provide biochemical and in vivo evidence that TRAF6 serves as an E3 to directly ubiquitinate NEMO. Reconstituting TRAF6 deficent cells with various TRAF6 mutants, we clearly demonstrate the requirement for the TRAF6 RING domain and site-specific auto-ubiquitination of TRAF6 to activate IKK in response to interleukin-1. These data establish a signaling cascade in which regulated site-specific Lys-63-linked TRAF6 auto-ubiquitination is the critical upstream mediator of IKK. PMID- 17135272 TI - Carbon monoxide protects against hyperoxia-induced endothelial cell apoptosis by inhibiting reactive oxygen species formation. AB - Hyperoxia causes cell injury and death associated with reactive oxygen species formation and inflammatory responses. Recent studies show that hyperoxia-induced cell death involves apoptosis, necrosis, or mixed phenotypes depending on cell type, although the underlying mechanisms remain unclear. Using murine lung endothelial cells, we found that hyperoxia caused cell death by apoptosis involving both extrinsic (Fas-dependent) and intrinsic (mitochondria-dependent) pathways. Hyperoxia-dependent activation of the extrinsic apoptosis pathway and formation of the death-inducing signaling complex required NADPH oxidase dependent reactive oxygen species production, because this process was attenuated by chemical inhibition, as well as by genetic deletion of the p47(phox) subunit, of the oxidase. Overexpression of heme oxygenase-1 prevented hyperoxia-induced cell death and cytochrome c release. Likewise, carbon monoxide, at low concentrations, markedly inhibited hyperoxia-induced endothelial cell death by inhibiting cytochrome c release and caspase-9/3 activation. Carbon monoxide, by attenuating hyperoxia-induced reactive oxygen species production, inhibited extrinsic apoptosis signaling initiated by death-inducing signal complex trafficking from the Golgi apparatus to the plasma membrane and downstream activation of caspase-8. We also found that carbon monoxide inhibited the hyperoxia-induced activation of Bcl-2-related proteins involved in both intrinsic and extrinsic apoptotic signaling. Carbon monoxide inhibited the activation of Bid and the expression and mitochondrial translocation of Bax, whereas promoted Bcl-X(L)/Bax interaction and increased Bad phosphorylation. We also show that carbon monoxide promoted an interaction of heme oxygenase-1 with Bax. These results define novel mechanisms underlying the antiapoptotic effects of carbon monoxide during hyperoxic stress. PMID- 17135273 TI - Crystal structure of the gamma-glutamyltranspeptidase precursor protein from Escherichia coli. Structural changes upon autocatalytic processing and implications for the maturation mechanism. AB - Gamma-glutamyltranspeptidase (GGT) is an extracellular enzyme that plays a key role in glutathione metabolism. The mature GGT is a heterodimer consisting of L- and S-subunits that is generated by posttranslational cleavage of the peptide bond between Gln-390 and Thr-391 in the precursor protein. Thr-391, which becomes the N-terminal residue of the S-subunit, acts as the active residue in the catalytic reaction. The crystal structure of a mutant GGT, T391A, that is unable to undergo autocatalytic processing, has been determined at 2.55-A resolution. Structural comparison of the precursor protein and mature GGT demonstrates that the structures of the core regions in the two proteins are unchanged, but marked differences are found near the active site. In particular, in the precursor, the segment corresponding to the C-terminal region of the L-subunit occupies the site where the loop (residues 438-449) forms the lid of the gamma-glutamyl group binding pocket in the mature GGT. This result demonstrates that, upon cleavage of the N-terminal peptide bond of Thr-391, the newly produced C terminus (residues 375-390) flips out, allowing the 438-449 segment to form the gamma-glutamyl group binding pocket. The electron density map for the T391A protein also identified a water molecule near the carbonyl carbon atom of Gln-390. The spatial arrangement around the water and Thr-391 relative to the scissile peptide bond appears suitable for the initiation of autocatalytic processing, as in other members of the N-terminal nucleophile hydrolase superfamily. PMID- 17135274 TI - The mixed-lineage leukemia fusion partner AF4 stimulates RNA polymerase II transcriptional elongation and mediates coordinated chromatin remodeling. AB - AF4 gene, frequently translocated with mixed-lineage leukemia (MLL) in childhood acute leukemia, encodes a putative transcriptional activator of the AF4/LAF4/FMR2 (ALF) protein family previously implicated in lymphopoiesis and Purkinje cell function in the cerebellum. Here, we provide the first evidence for a direct role of AF4 in the regulation of transcriptional elongation by RNA polymerase II (Pol II). We demonstrate that mouse Af4 functions as a positive regulator of Pol II transcription elongation factor b (P-TEFb) kinase and, in complex with MLL fusion partners Af9, Enl and Af10, as a mediator of histone H3-K79 methylation by recruiting Dot1 to elongating Pol II. These pathways are interconnected and tightly regulated by the P-TEFb-dependent phosphorylation of Af4, Af9 and Enl which controls their transactivation activity and/or protein stability. Consistently, increased levels of phosphorylated Pol II and methylated H3-K79 are observed in the ataxic mouse mutant robotic, an over-expression model of Af4. Finally, we confirm the functional relevance of Af4, Enl and Af9 to the regulation of gene transcription as their over-expression strongly stimulates P TEFb-dependent transcription of a luciferase reporter gene. Our findings uncover a central role for these proteins in the regulation of transcriptional elongation and coordinated histone methylation, providing valuable insight into their contribution to leukemogenesis and neurodegeneration. Since these activities likely extend to the entire ALF protein family, this study also significantly inputs our understanding of the molecular basis of FRAXE mental retardation syndrome in which FMR2 expression is silenced. PMID- 17135275 TI - Involvement of hyperprolinemia in cognitive and psychiatric features of the 22q11 deletion syndrome. AB - Microdeletions of the 22q11 region, responsible for the velo-cardio-facial syndrome (VCFS), are associated with an increased risk for psychosis and mental retardation. Recently, it has been shown in a hyperprolinemic mouse model that an interaction between two genes localized in the hemideleted region, proline dehydrogenase (PRODH) and catechol-o-methyl-transferase (COMT), could be involved in this phenotype. Here, we further characterize in eight children the molecular basis of type I hyperprolinemia (HPI), a recessive disorder resulting from reduced activity of proline dehydrogenase (POX). We show that these patients present with mental retardation, epilepsy and, in some cases, psychiatric features. We next report that, among 92 adult or adolescent VCFS subjects, a subset of patients with severe hyperprolinemia has a phenotype distinguishable from that of other VCFS patients and reminiscent of HPI. Forward stepwise multiple regression analysis selected hyperprolinemia, psychosis and COMT genotype as independent variables influencing IQ in the whole VCFS sample. An inverse correlation between plasma proline level and IQ was found. In addition, as predicted from the mouse model, hyperprolinemic VCFS subjects bearing the Met COMT low activity allele are at risk for psychosis (OR = 2.8, 95% CI = 1.04-7.4). Finally, from the extensive analysis of the PRODH gene coding sequence variations, it is predicted that POX residual activity in the 0-30% range results into HPI, whereas residual activity in the 30-50% range is associated either with normal plasma proline levels or with mild-to-moderate hyperprolinemia. PMID- 17135276 TI - APC mutations in FAP-associated desmoid tumours are non-random but not 'just right'. AB - Analysis of APC mutations in colonic and duodenal tumours from familial adenomatous polyposis (FAP) patients has shown that the site of the first hit, the germline mutation, can predict the type and position of the somatic mutation or 'second hit'. The two APC mutations are selected on the basis of a 'just right' level of beta-catenin signalling in intestinal tumours achieved through retention of some of the seven 20-amino-acid beta-catenin degradation repeats. Desmoids are a life threatening extra-colonic manifestation in FAP patients. These aggressive tumours of mesenchymal origin are, at present, poorly characterized in terms of mutational APC spectra. We have investigated somatic mutations in the largest cohort of FAP-associated desmoids to date, and combined our results with previously published data. Somatic mutations were found to occur non-randomly and the position of the germline mutation shown to be a major determinant of the somatic mutation, a characteristic shared with intestinal tumours from FAP patients. In contrast to colonic polyps, loss of heterozygosity in desmoids involved deletion rather than mitotic recombination. While tumours from the colorectum and upper gastrointestinal tract usually retain one to two and three to four beta-catenin degradation repeats, respectively, most desmoids preferentially retain two repeats (P < 0.001, chi2 test). In addition, most desmoids with two APC hits (87%, 26/30) had one mutated allele with no 20-amino acid repeats (P < 0.001). This feature, unique among FAP tumours, indicates that a mutation deleting all repeats from one allele may be an important component in maintaining appropriate levels of beta-catenin signalling levels in desmoid tumour cells. PMID- 17135277 TI - The first 17 amino acids of Huntingtin modulate its sub-cellular localization, aggregation and effects on calcium homeostasis. AB - A truncated form of the Huntington's disease (HD) protein that contains the polyglutamine repeat, Httex1p, causes HD-like phenotypes in multiple model organisms. Molecular signatures of pathogenesis appear to involve distinct domains within this polypeptide. We studied the contribution of each domain, singly or in combination, to sub-cellular localization, aggregation and intracellular Ca2+ ([Ca2+]i) dynamics in cells. We demonstrate that sub-cellular localization is most strongly influenced by the first 17 amino acids, with this sequence critically controlling Httex1p mitochondrial localization and also promoting association with the endoplasmic reticulum (ER) and Golgi. This domain also enhances the formation of visible aggregates and together with the expanded polyQ repeat acutely disrupts [Ca2+]i levels in glutamate-challenged PC12 cells. Isolated cortical mitochondria incubated with Httex1p resulted in uncoupling and depolarization of these organelles, further supporting the idea that Httex1p dependent mitochondrial dysfunction could be instrumental in promoting acute Ca2+ dyshomeostasis. Interestingly, neither mitochondrial nor ER associations seem to be required to promote long-term [Ca2+]i dyshomeostasis. PMID- 17135278 TI - Cholinergic nicotinic receptor genes implicated in a nicotine dependence association study targeting 348 candidate genes with 3713 SNPs. AB - Nicotine dependence is one of the world's leading causes of preventable death. To discover genetic variants that influence risk for nicotine dependence, we targeted over 300 candidate genes and analyzed 3713 single nucleotide polymorphisms (SNPs) in 1050 cases and 879 controls. The Fagerstrom test for nicotine dependence (FTND) was used to assess dependence, in which cases were required to have an FTND of 4 or more. The control criterion was strict: control subjects must have smoked at least 100 cigarettes in their lifetimes and had an FTND of 0 during the heaviest period of smoking. After correcting for multiple testing by controlling the false discovery rate, several cholinergic nicotinic receptor genes dominated the top signals. The strongest association was from an SNP representing CHRNB3, the beta3 nicotinic receptor subunit gene (P = 9.4 x 10( 5)). Biologically, the most compelling evidence for a risk variant came from a non-synonymous SNP in the alpha5 nicotinic receptor subunit gene CHRNA5 (P = 6.4 x 10(-4)). This SNP exhibited evidence of a recessive mode of inheritance, resulting in individuals having a 2-fold increase in risk of developing nicotine dependence once exposed to cigarette smoking. Other genes among the top signals were KCNJ6 and GABRA4. This study represents one of the most powerful and extensive studies of nicotine dependence to date and has found novel risk loci that require confirmation by replication studies. PMID- 17135279 TI - The DYRK1A gene, encoded in chromosome 21 Down syndrome critical region, bridges between beta-amyloid production and tau phosphorylation in Alzheimer disease. AB - We scanned throughout chromosome 21 to assess genetic associations with late onset Alzheimer disease (AD) using 374 Japanese patients and 375 population-based controls, because trisomy 21 is known to be associated with early deposition of beta-amyloid (Abeta) in the brain. Among 417 markers spanning 33 Mb, 22 markers showed associations with either the allele or the genotype frequency (P < 0.05). Logistic regression analysis with age, sex and apolipoprotein E (APOE)-epsilon4 dose supported genetic risk of 17 markers, of which eight markers were linked to the SAMSN1, PRSS7, NCAM2, RUNX1, DYRK1A and KCNJ6 genes. In logistic regression, the DYRK1A (dual-specificity tyrosine-regulated kinase 1A) gene, located in the Down syndrome critical region, showed the highest significance [OR = 2.99 (95% CI: 1.72-5.19), P = 0.001], whereas the RUNX1 gene showed a high odds ratio [OR = 23.3 (95% CI: 2.76-196.5), P = 0.038]. DYRK1A mRNA level in the hippocampus was significantly elevated in patients with AD when compared with pathological controls (P < 0.01). DYRK1A mRNA level was upregulated along with an increase in the Abeta-level in the brain of transgenic mice, overproducing Abeta at 9 months of age. In neuroblastoma cells, Abeta induced an increase in the DYRK1A transcript, which also led to tau phosphorylation at Thr212 under the overexpression of tau. Therefore, the upregulation of DYRK1A transcription results from Abeta loading, further leading to tau phosphorylation. Our result indicates that DYRK1A could be a key molecule bridging between beta-amyloid production and tau phosphorylation in AD. PMID- 17135280 TI - Growth retardation, DNA repair defects, and lack of spermatogenesis in BRCA1 deficient mice. PMID- 17135281 TI - Neointimal coverage of sirolimus-eluting stents at 6-month follow-up: evaluated by optical coherence tomography. AB - AIMS: Since the intravascular ultrasound (IVUS) cannot detect neointimal layers in the majority of sirolimus-eluting stents (SES) at the chronic phase, it is still controversial to what extent SES remain uncovered. However, optical coherence tomography (OCT) with excellent resolution may be able to detect thinner neointima. METHODS AND RESULTS: A total of 34 patients (57 SES) underwent OCT and IVUS evaluations at 6-month follow-up. The thickness of neointima on each SES strut cross-section and strut apposition to the vessel wall was evaluated. By OCT evaluation, the median (25th, 75th percentiles) neointima thickness was 52.5 microm (28.0 microm, 147.6 microm) and the prevalence of struts covered by thin neointima undetectable by IVUS was 64%. The average rate of neointima-covered struts in an individual SES was 89%. Nine SES (16%) showed full coverage by neointima, whereas the remaining stents had partially uncovered strut lesions. Among the 6840 struts visualized by OCT in all of the SES, 79 struts showed malapposition without neointimal coverage, and were frequently observed in the areas of SES overlap. CONCLUSION: At 6 months, most of the SES were covered with thin neointima, but few showed full coverage. PMID- 17135282 TI - The long-term value of sirolimus- and paclitaxel-eluting stents over bare metal stents in patients with diabetes mellitus. AB - AIMS: To investigate the outcome of a real world diabetic patient cohort treated with bare metal stents (BMS), sirolimus-, or paclitaxel-eluting stents (SES and PES, respectively). Due to the different mechanisms of action of both drugs it is currently unknown which device is the best option to treat these high-risk patients. METHODS AND RESULTS: The study compares the 2-year clinical outcome of 708 consecutive diabetic patients (25% insulin treated) treated with either a BMS (n = 252), a SES (n = 206), or a PES (n = 250), as part of the RESEARCH and T SEARCH registries. Target vessel revascularization was 19.5% in the BMS group, vs. 15.3% in the SES group and 9.7% in the PES group. PES (21.2%), but not SES (28.9%), were superior to BMS (29.7%) in reducing major adverse cardiac events. After propensity analyses, none of the differences remained significant. The incidence of stent thrombosis (ST) was high in both DES groups. CONCLUSION: There was a trend towards a more favourable outcome associated with the use of PES over BMS. There was no significant difference between SES and PES in each of the clinical endpoints, and neither in the NIDDM patients, which are hypothesized to be better-off with PES. PMID- 17135283 TI - Increased risk of acute myocardial infarction and elevated levels of C-reactive protein in carriers of the Thr-87 variant of the ATP receptor P2Y11. AB - AIMS: Extracellular ATP acting on the P2Y11 receptor regulates inflammatory cells. We hypothesized that polymorphisms in the receptor could influence the risk of acute myocardial infarction (AMI). METHODS AND RESULTS: In the Malmo diet and cancer AMI case-control study (n = 3732) the P2Y11 gene Thr-87 polymorphism was present in 19.8% of the controls and 22.9% in AMI patients (OR 1.21; P = 0.03). Stronger associations were found in patients with family history (FH) of AMI, 1.32; early-onset (EO) AMI, 1.43; or EO AMI combined with FH, 1.50; supporting a genetic mechanism. The Thr-87 homozygotes had an even greater risk of AMI, 1.94 (P = 0.04); and 2.48 in the EO AMI subgroup, suggesting a genetic dosage effect. In the cardiovascular risk factor group (n = 6055), 21.3% carried the Thr-87 allele. C-reactive protein was elevated in Thr-87 carriers: 1.6 mg/L vs. 1.3 mg/L (P = 0.001). No difference was seen for blood pressure, lipids, body mass index, smoking, or diabetes mellitus. CONCLUSION: The common Ala-87-Thr polymorphism of the P2Y11 receptor is associated with AMI and increased levels of C-reactive protein. We hypothesize that an inflammatory mechanism might be involved. The P2Y11 receptor is a promising new drug target in the prevention of AMI. PMID- 17135284 TI - Reduction in infarct size, but no functional improvement after bone marrow cell administration in a porcine model of reperfused myocardial infarction. AB - AIMS: Stem cell therapy after myocardial infarction (MI) has been studied in models of permanent coronary occlusion. We studied the effect of intracoronary administration of unselected bone marrow (BM) and mononuclear cells (MNC) in a porcine model of reperfused MI. METHODS AND RESULTS: In 34 swine, the left circumflex coronary artery was balloon-occluded for 2 h followed by reperfusion. Ten swine without MI served as controls. All swine underwent magnetic resonance imaging (MRI) 1 week post-MI. The next day, 10 of the 30 surviving MI swine received BM, 10 other MI swine received MNC, and the remaining MI swine received medium intracoronary. Four weeks later, all swine underwent a follow-up MRI. One week after MI, end-diastolic volume (92+/-16 mL) and left ventricular (LV) weight (78+/-12 g) were greater, whereas ejection fraction (40+/-8%) was lower than in controls (69+/-11 mL, 62+/-13 g, and 53+/-6%). Injection of BM or MNC had no effect on the MI-induced changes in global or regional LV-function. However, there was a significant reduction in infarct size 4 weeks after MNC injection ( 6+/-3%) compared with the medium (-3+/-5%). CONCLUSION: Intracoronary injection of BM or MNC in swine does not improve regional or global LV-function 4 weeks after injection. However, a reduction in infarct-size was noted after MNC injection. PMID- 17135285 TI - Estrogen, cognition and female ageing. AB - Starting from fetal life, estrogens are crucial in determining central gender dimorphism, and an estrogen-induced synaptic plasticity is well evident during puberty and seasonal changes as well as during the ovarian cycle. Estrogens act on the central nervous system (CNS) both through genomic mechanisms, modulating synthesis, release and metabolism of neurotransmitters, neuropeptides and neurosteroids, and through non-genomic mechanisms, influencing electrical excitability, synaptic function and morphological features. Therefore, estrogen's neuroactive effects are multifaceted and encompass a system that ranges from the chemical to the biochemical to the genomic mechanisms, protecting against a wide range of neurotoxic insults. Clinical evidences show that, during the climacteric period, estrogen withdrawal in the limbic system gives rise to modifications in mood, behaviour and cognition and that estrogen administration is able to improve mood and cognitive efficiency in post-menopause. Many biological mechanisms support the hypothesis that estrogens might protect against Alzheimer's disease (AD) by influencing neurotransmission, increasing cerebral blood flow, modulating growth proteins associated with axonal elongation and blunting the neurotoxic effects of beta-amyloid. On the contrary, clinical studies of estrogen replacement therapy (ERT) and cognitive function have reported controversial results, indicating a lack of efficacy of estrogens on cognition in post menopausal women aged >or=65 years. These findings suggest the presence of a critical period for HRT-related neuroprotection and underlie the potential importance of early initiation of therapy for cognitive benefit. In this review, we shall first describe the multiple effects of steroids in the nervous system, which may be significant in the ageing process. A critical update of HRT use in women and a discussion of possible prospectives for steroid use are subsequently proposed. PMID- 17135286 TI - Protophloem differentiation in early Arabidopsis thaliana development. AB - During Arabidopsis embryogenesis, procambial cells undergo coordinated, asymmetric cell divisions, giving rise to vascular precursor cells (protophloem and protoxylem precursors). After germination, these cells terminally differentiate into specialized conducting cells, referred to as protophloem and protoxylem cells. Few readily identifiable markers of the onset of specification and differentiation are available, hampering the molecular genetic analysis of protophloem development. Confocal microscopy was used to investigate the patterning and differentiation of phloem cells during early plant development. Longitudinal divisions of phloem initials allowed the identification of protophloem precursor cells and adjacent metaphloem initials along the length of the plant. During germination, protophloem differentiation was observed at two independent locations, in the cotyledons and the hypocotyl. In both locations, differentiation was concomitant with cell elongation. We identified five gene trap lines (PD1-PD5) with marker gene expression in immature protophloem elements. The spatio-temporal marker expression pattern of the lines divides them into two groups. The early specification markers PD4 and PD5 were expressed in developing organs before procambium formation and then became restricted to phloem initial cells. The protophloem precursor markers PD1-PD3 were expressed in differentiating protophloem cells at different stages of their development. All markers were expressed transiently and iteratively during the differentiation of protophloem in newly formed organs. Flanking genes were identified for four out of five gene-trap insertion lines. The possible function of these genes with respect to phloem differentiation is discussed. PMID- 17135287 TI - The assembly pathway of the 19S regulatory particle of the yeast 26S proteasome. AB - The 26S proteasome consists of the 20S proteasome (core particle) and the 19S regulatory particle made of the base and lid substructures, and it is mainly localized in the nucleus in yeast. To examine how and where this huge enzyme complex is assembled, we performed biochemical and microscopic characterization of proteasomes produced in two lid mutants, rpn5-1 and rpn7-3, and a base mutant DeltaN rpn2, of the yeast Saccharomyces cerevisiae. We found that, although lid formation was abolished in rpn5-1 mutant cells at the restrictive temperature, an apparently intact base was produced and localized in the nucleus. In contrast, in DeltaN rpn2 cells, a free lid was formed and localized in the nucleus even at the restrictive temperature. These results indicate that the modules of the 26S proteasome, namely, the core particle, base, and lid, can be formed and imported into the nucleus independently of each other. Based on these observations, we propose a model for the assembly process of the yeast 26S proteasome. PMID- 17135288 TI - Prohibitins interact genetically with Atp23, a novel processing peptidase and chaperone for the F1Fo-ATP synthase. AB - The generation of cellular energy depends on the coordinated assembly of nuclear and mitochondrial-encoded proteins into multisubunit respiratory chain complexes in the inner membrane of mitochondria. Here, we describe the identification of a conserved metallopeptidase present in the intermembrane space, termed Atp23, which exerts dual activities during the biogenesis of the F(1)F(O)-ATP synthase. On one hand, Atp23 serves as a processing peptidase and mediates the maturation of the mitochondrial-encoded F(O)-subunit Atp6 after its insertion into the inner membrane. On the other hand and independent of its proteolytic activity, Atp23 promotes the association of mature Atp6 with Atp9 oligomers. This assembly step is thus under the control of two substrate-specific chaperones, Atp10 and Atp23, which act on opposite sides of the inner membrane. Strikingly, both ATP10 and ATP23 were found to genetically interact with prohibitins, which build up large, ring-like assemblies with a proposed scaffolding function in the inner membrane. Our results therefore characterize not only a novel processing peptidase with chaperone activity in the mitochondrial intermembrane space but also link the function of prohibitins to the F(1)F(O)-ATP synthase complex. PMID- 17135289 TI - Aberrant translation of cytochrome c oxidase subunit 1 mRNA species in the absence of Mss51p in the yeast Saccharomyces cerevisiae. AB - Expression of yeast mitochondrial genes depends on specific translational activators acting on the 5'-untranslated region of their target mRNAs. Mss51p is a translational factor for cytochrome c oxidase subunit 1 (COX1) mRNA and a key player in down-regulating Cox1p expression when subunits with which it normally interacts are not available. Mss51p probably acts on the 5'-untranslated region of COX1 mRNA to initiate translation and on the coding sequence itself to facilitate elongation. Mss51p binds newly synthesized Cox1p, an interaction that could be necessary for translation. To gain insight into the different roles of Mss51p on Cox1p biogenesis, we have analyzed the properties of a new mitochondrial protein, mp15, which is synthesized in mss51 mutants and in cytochrome oxidase mutants in which Cox1p translation is suppressed. The mp15 polypeptide is not detected in cox14 mutants that express Cox1p normally. We show that mp15 is a truncated translation product of COX1 mRNA whose synthesis requires the COX1 mRNA-specific translational activator Pet309p. These results support a key role for Mss51p in translationally regulating Cox1p synthesis by the status of cytochrome oxidase assembly. PMID- 17135290 TI - The metalloprotease encoded by ATP23 has a dual function in processing and assembly of subunit 6 of mitochondrial ATPase. AB - In the present study we have identified a new metalloprotease encoded by the nuclear ATP23 gene of Saccharomyces cerevisiae that is essential for expression of mitochondrial ATPase (F(1)-F(O) complex). Mutations in ATP23 cause the accumulation of the precursor form of subunit 6 and prevent assembly of F(O). Atp23p is associated with the mitochondrial inner membrane and is conserved from yeast to humans. A mutant harboring proteolytically inactive Atp23p accumulates the subunit 6 precursor but is nonetheless able to assemble a functional ATPase complex. These results indicate that removal of the subunit 6 presequence is not an essential event for ATPase biogenesis and that Atp23p, in addition to its processing activity, must provide another important function in F(O) assembly. The product of the yeast ATP10 gene was previously shown to interact with subunit 6 and to be required for its association with the subunit 9 ring. In this study one extra copy of ATP23 was found to be an effective suppressor of an atp10 null mutant, suggesting an overlap in the functions of Atp23p and Atp10p. Atp23p may, therefore, also be a chaperone, which in conjunction with Atp10p mediates the association of subunit 6 with the subunit 9 ring. PMID- 17135291 TI - Atg27 is required for autophagy-dependent cycling of Atg9. AB - Autophagy is a catabolic pathway for the degradation of cytosolic proteins or organelles and is conserved among all eukaryotic cells. The hallmark of autophagy is the formation of double-membrane cytosolic vesicles, termed autophagosomes, which sequester cytoplasm; however, the mechanism of vesicle formation and the membrane source remain unclear. In the yeast Saccharomyces cerevisiae, selective autophagy mediates the delivery of specific cargos to the vacuole, the analog of the mammalian lysosome. The transmembrane protein Atg9 cycles between the mitochondria and the pre-autophagosomal structure, which is the site of autophagosome biogenesis. Atg9 is thought to mediate the delivery of membrane to the forming autophagosome. Here, we characterize a second transmembrane protein Atg27 that is required for specific autophagy in yeast. Atg27 is required for Atg9 cycling and shuttles between the pre-autophagosomal structure, mitochondria, and the Golgi complex. These data support a hypothesis that multiple membrane sources supply the lipids needed for autophagosome formation. PMID- 17135292 TI - Efficient cargo sorting by ESCRT-I and the subsequent release of ESCRT-I from multivesicular bodies requires the subunit Mvb12. AB - The endosomal sorting complex required for transport (ESCRT)-I protein complex functions in recognition and sorting of ubiquitinated transmembrane proteins into multivesicular body (MVB) vesicles. It has been shown that ESCRT-I contains the vacuolar protein sorting (Vps) proteins Vps23, Vps28, and Vps37. We identified an additional subunit of yeast ESCRT-I called Mvb12, which seems to associate with ESCRT-I by binding to Vps37. Transient recruitment of ESCRT-I to MVBs results in the rapid degradation of Mvb12. In contrast to mutations in other ESCRT-I subunits, which result in strong defects in MVB cargo sorting, deletion of MVB12 resulted in only a partial sorting phenotype. This trafficking defect was fully suppressed by overexpression of the ESCRT-II complex. Mutations in MVB12 did not affect recruitment of ESCRT-I to MVBs, but they did result in delivery of ESCRT-I to the vacuolar lumen via the MVB pathway. Together, these observations suggest that Mvb12 may function in regulating the interactions of ESCRT-I with cargo and other proteins of the ESCRT machinery to efficiently coordinate cargo sorting and release of ESCRT-I from the MVB. PMID- 17135293 TI - A chemoattractant-mediated Gi-coupled pathway activates adenylyl cyclase in human neutrophils. AB - Neutrophils and Dictyostelium use conserved signal transduction pathways to decipher chemoattractant gradients and migrate directionally. In both cell types, addition of chemoattractants stimulates the production of cAMP, which has been suggested to regulate chemotaxis. We set out to define the mechanism by which chemoattractants increase cAMP levels in human neutrophils. We show that chemoattractants elicit a rapid and transient activation of adenylyl cyclase (AC). This activation is sensitive to pertussis toxin treatment but independent of phosphoinositide-3 kinase activity and an intact cytoskeleton. Remarkably, and in sharp contrast to Galpha(s)-mediated activation, chemoattractant-induced AC activation is lost in cell lysates. Of the nine, differentially regulated transmembrane AC isoforms in the human genome, we find that isoforms III, IV, VII, and IX are expressed in human neutrophils. We conclude that the signal transduction cascade used by chemoattractants to activate AC is conserved in Dictyostelium and human neutrophils and is markedly different from the canonical Galpha(s)-meditated pathway. PMID- 17135294 TI - Modifications of cellular responses to lysophosphatidic acid and platelet activating factor by plasma gelsolin. AB - Gelsolin is a highly conserved intracellular actin-binding protein with an extracellular isoform, plasma gelsolin (pGSN). Blood concentrations of pGSN decrease in response to diverse tissue injuries. Depletion of pGSN to critical levels precedes and often predicts complications of injuries such as lung permeability changes and death. Administration of recombinant pGSN ameliorates such complications and reduces mortality in animal models. One proposed mechanism for pGSN's protective effects is that it inhibits inflammatory mediators generated during primary injuries, since pGSN binds bioactive mediators, including lysophospatidic acid (LPA) and endotoxin in vitro. However, no direct evidence in support of this hypothesis has been available. Here we show that recombinant pGSN modestly inhibited LPA-induced P-selectin upregulation by human platelets in the presence of albumin (P < 0.0001). However, physiologically relevant pGSN concentrations inhibit platelet-activating factor (PAF)-mediated P selectin expression by up to 77% (P < 0.0001). pGSN also markedly inhibited PAF induced superoxide anion (O(2)(-)) production of human peripheral neutrophils (PMN) in a concentration-dependent manner (P < 0.0001). A phospholipid-binding peptide derived from pGSN (QRLFQVKGRR) also inhibited PAF-mediated O(2)(-) generation (P = 0.024). Therefore, pGSN interferes with PAF- and LPA-induced cellular activation in vitro, suggesting a mechanism for the protective role of pGSN in vivo. PMID- 17135295 TI - Phosphorothioate oligonucleotides reduce mitochondrial outer membrane permeability to ADP. AB - G3139, an antisense Bcl-2 phosphorothioate oligodeoxyribonucleotide, induces apoptosis in melanoma and other cancer cells. This apoptosis happens before and in the absence of the downregulation of Bcl-2 and thus seems to be Bcl-2 independent. Binding of G3139 to mitochondria and its ability to close voltage dependent anion-selective channel (VDAC) have led to the hypothesis that G3139 acts, in part, by interacting with VDAC channels in the mitochondrial outer membrane (21). In this study, we demonstrate that G3139 is able to reduce the mitochondrial outer membrane permeability to ADP by a factor of 6 or 7 with a K(i) between 0.2 and 0.5 microM. Because VDAC is responsible for this permeability, this result strengthens the aforesaid hypothesis. Other mitochondrial respiration components are not affected by [G3139] up to 1 microM. Higher levels begin to inhibit respiration rates, decrease light scattering and increase uncoupled respiration. These results agree with accumulating evidence that VDAC closure favors cytochrome c release. The speed of this effect (within 10 min) places it early in the apoptotic cascade with cytochrome c release occurring at later times. Other phosphorothioate oligonucleotides are also able to induce VDAC closure, and there is some length dependence. The phosphorothioate linkages are required to induce the reduction of outer membrane permeability. At levels below 1 microM, phosphorothioate oligonucleotides are the first specific tools to restrict mitochondrial outer membrane permeability. PMID- 17135296 TI - Oligomeric structure of the neutral amino acid transporters KAAT1 and CAATCH1. AB - The highly homologous neutral amino acid transporters KAAT1 and CAATCH1, cloned from the midgut epithelium of the Manduca sexta larva, are members of the Na(+)/Cl(-)-dependent transporter family. Recent evidence indicates that transporters of this family form constitutive oligomers. CAATCH1 and KAAT1 give rise to specific kinds of current depending on the transported amino acid, cotransported ion, pH, and membrane voltage. Different substrates induce notably distinct transport-associated currents in the two proteins that represent useful tools in structural-functional studies. To determine whether KAAT1 and CAATCH1 form functional oligomers, we have constructed four concatameric proteins for electrophysiological analysis, consisting of one KAAT1 protein covalently linked to another KAAT1 (K-K concatamer) or to CAATCH1 (K-C concatamer) and vice versa (C-C concatamer and C-K concatamer), and eight constructs where the two transporters were linked to yellow or cyan fluorescent protein in the NH(2) or COOH terminus, to determine the oligomer formation and the relative distance between the different subunits by fluorescence resonance energy transfer (FRET) analysis. Heterologous expression of the concatenated constructs and coinjection of the original proteins in different proportions allowed us to compare the characteristics of the currents to those of the oocytes expressing only the wild type proteins. All the constructs were fully active, and their electrophysiological behavior was consistent with the activity as monomeric proteins. However, the FRET studies indicate that these transporters form oligomers in agreement with the LeuT(Aa) atomic structure and confirm that the COOH termini of the adjacent subunits are closer than NH(2) termini. PMID- 17135297 TI - Impact of Type 2 diabetes and aging on cardiomyocyte function and O-linked N acetylglucosamine levels in the heart. AB - Increased levels of O-linked attachment of N-acetylglucosamine (O-GlcNAc) on nucleocytoplasmic proteins are implicated in the development of diabetic cardiomyopathy and are regulated by O-GlcNAc transferase (OGT) expression and its substrate UDP-GlcNAc. Therefore, the goal of this study was to determine whether the development of diabetes in the Zucker diabetic fatty (ZDF) rat, a model of Type 2 diabetes, results in defects in cardiomyocyte mechanical function and, if so, whether this is associated with increased levels of O-GlcNAc and increased OGT expression. Six-week-old ZDF rats were hyperinsulinemic but normoglycemic, and there were no differences in cardiomyocyte mechanical function, UDP-GlcNAc, O GlcNAc, or OGT compared with age-matched lean control rats. Cardiomyocytes isolated from 22-wk-old hyperglycemic ZDF rats exhibited significantly impaired relaxation, compared with both age-matched lean control and 6-wk-old ZDF groups. There was also a significant increase in O-GlcNAc levels in high-molecular-mass proteins in the 22-wk-old ZDF group compared with age-matched lean control and 6 wk-old ZDF groups; this was associated with increased UDP-GlcNAc levels but not increased OGT expression. Surprisingly, there was a significant decrease in overall O-GlcNAc levels between 6 and 22 wk of age in lean, ZDF, and Sprague Dawley rats that was associated with decreased OGT expression. These results support the notion that an increase in O-GlcNAc on specific proteins may contribute to impaired cardiomyocyte function in diabetes. However, this study also indicates that in the heart the level of O-GlcNAc on proteins appears to be differentially regulated by age and diabetes. PMID- 17135298 TI - TGF-beta-regulated collagen type I accumulation: role of Src-based signals. AB - Transforming growth factor-beta (TGF-beta) stimulates myofibroblast transdifferentiation, leading to type I collagen accumulation and fibrosis. We investigated the function of Src in TGF-beta-induced collagen I accumulation. In human mesangial cells, PTyr416 Src (activated Src) was 3.3-fold higher in TGF beta-treated cells than in controls. Src activation by TGF-beta was blocked by rottlerin and by a dominant negative mutant of protein kinase Cdelta (PKCdelta), showing that TGF-beta activates Src by a PKCdelta-based mechanism. Pharmacological inhibitors and a dominant negative Src mutant prevented the increase in collagen type I secretion in cells exposed to TGF-beta. Similarly, on target Src small interference RNA (siRNA) prevented type I collagen secretion in response to TGF-beta, but off-target siRNA complexes had no effect. It is well established in mesangial cells that upregulation of type I collagen by TGF-beta requires extracellular signal-regulated kinase 1/2 (ERK1/2), and we found that activation of ERK1/2 by TGF-beta requires Src. In conclusion, these results suggest that stimulation of collagen type I secretion by TGF-beta requires a PKCdelta-Src-ERK1/2 signaling motif. PMID- 17135299 TI - Regulation of the human biotin transporter hSMVT promoter by KLF-4 and AP-2: confirmation of promoter activity in vivo. AB - The mechanism of biotin uptake in human intestine has been well characterized and involves the human sodium-dependent multivitamin transporter (hSMVT), yet little is known about the molecular/transcriptional regulation of the system. Previous investigations cloned the 5' regulatory region of the hSMVT gene and identified the minimal promoter. To expand these investigations, we compared activity of the hSMVT promoter in three human intestinal epithelial cell lines (NCM460, Caco-2, and HuTu-80) and contrasted a renal epithelial cell line (HEK-293). We analyzed the role of putative cis-elements in regulating promoter activity and confirmed activity of the cloned hSMVT promoter in vivo. In vitro studies demonstrated that all cell lines utilized the same minimal promoter region, and mutation of specific cis-regulatory elements [Kruppel-like factor 4 (KLF-4) and activator protein-2 (AP-2)] led to a decrease in promoter activity in all intestinal cell types but not in renal cells. Using electrophoretic mobility shift assays, we identified two specific DNA/protein complexes. Using oligonucleotide competition and antibody supershift analysis, we determined that KLF-4 and AP-2 were involved in forming the complexes. In HEK-293 cells, overexpressing KLF-4 increased the endogenous hSMVT message levels threefold and activated a cotransfected hSMVT promoter-reporter construct. In vivo studies using hSMVT promoter-luciferase transgenic mice established physiological relevance and showed the pattern of hSMVT promoter expression to be similar to endogenous mouse SMVT mRNA expression. The results demonstrate, for the first time, the importance of KLF-4 and AP-2 in regulating the activity of the hSMVT promoter in the intestine and provide direct in vivo confirmation of hSMVT promoter activity. PMID- 17135300 TI - Chloride channelopathy in myotonic dystrophy resulting from loss of posttranscriptional regulation for CLCN1. AB - Transmembrane chloride ion conductance in skeletal muscle increases during early postnatal development. A transgenic mouse model of myotonic dystrophy type 1 (DM1) displays decreased sarcolemmal chloride conductance. Both effects result from modulation of chloride channel 1 (CLCN1) expression, but the respective contributions of transcriptional vs. posttranscriptional regulation are unknown. Here we show that alternative splicing of CLCN1 undergoes a physiological splicing transition during the first 3 wk of postnatal life in mice. During this interval, there is a switch to production of CLCN1 splice products having an intact reading frame, an upregulation of CLCN1 mRNA encoding full-length channel protein, and an increase of CLCN1 function, as determined by patch-clamp analysis of single muscle fibers. In a transgenic mouse model of DM1, however, the splicing transition does not occur, CLCN1 channel function remains low throughout the postnatal interval, and muscle fibers display myotonic discharges. Thus alternative splicing is a posttranscriptional mechanism regulating chloride conductance during muscle development, and the chloride channelopathy in a transgenic mouse model of DM1 results from a failure to execute a splicing transition for CLCN1. PMID- 17135301 TI - Focal adhesion kinase mediates cell survival via NF-kappaB and ERK signaling pathways. AB - Focal adhesion kinase (FAK) is important to cellular functions such as proliferation, migration, and survival of anchorage-dependent cells. We investigated the role of FAK in modulating normal cellular responses, specifically cell survival in response to inflammatory stimuli and serum withdrawal, using FAK-knockout (FAK(-/-)) embryonic fibroblasts. FAK(-/-) fibroblasts were more vulnerable to TNF-alpha-induced apoptosis, as measured by terminal deoxynucleotidyl transferase positivity. FAK(-/-) fibroblasts also demonstrated increased procaspase-3 cleavage to p17 subunit, whereas this was undetectable in FAK(+/+) fibroblasts. Insulin receptor substrate-1 expression was completely abolished and NF-kappaB activity was reduced, with a concomitant decrease in abundance of the anti-apoptotic protein Bcl-x(L) in FAK(-/-) cells. Upon serum withdrawal, FAK(+/+) cells exhibited marked attenuation of basal ERK phosphorylation, while FAK(-/-) cells, in contrast, maintained high basal ERK phosphorylation. Moreover, inhibition of ERK phosphorylation potentiated serum withdrawal-induced caspase-3 activity. This was paralleled by increased insulin receptor substrate (IRS)-2 expression in FAK(-/-) cells, although both insulin- and IGF-1-mediated phosphorylation of Akt/PKB and GSK-3 were impaired. This suggests that IRS-2 protects against apoptosis upon serum withdrawal via the ERK signaling pathway. The specific role of FAK to protect cells from apoptosis is regulated by activation and phosphorylation of NF-kappaB and interaction between activated growth factor anti-apoptotic signaling pathways involving both phosphatidylinositol 3-kinase/Akt and MAPK/ERK1/2. We demonstrate that FAK is necessary for upregulation of the anti-apoptotic NF-kappaB response, as well as for normal expression of growth factor signaling proteins. Thus we propose a novel role for FAK in protection from cytokine-mediated apoptosis. PMID- 17135302 TI - ROS and NF-kappaB but not LXR mediate IL-1beta signaling for the downregulation of ATP-binding cassette transporter A1. AB - ATP-binding cassette transporter A1 (ABCA1), a pivotal regulator of cholesterol efflux from cells to apolipoproteins, plays an important role in cholesterol homeostasis. As an inflammatory factor, IL-1beta has been shown to downregulate ABCA1 in macrophages and facilitates foam cell formation. However, the molecular mechanism underlining the downregulated ABCA1 by IL-1beta is still elusive. In the present study, we demonstrated that IL-1beta downregulated ABCA1 but not ABCG1 at mRNA and protein levels in a time- and dose-dependent manner in THP-1 and A549 cells. IL-1beta attenuated ABCA1 promoter activity through an LXR (liver X receptor)-independent pathway, since IL-1beta did not alter the expression and activities of LXRalpha/beta, and deletion of the LXR responsive element from the ABCA1 promoter failed to reverse the IL-1beta effect. In contrast, NF-kappaB inhibition by pyrrolidine dithiocarbamate and MG132 prevented the suppression of ABCA1 by IL-1beta. Cotransfection with ABCA1 luciferase reporter and the expression plasmids of Rel A decreased ABCA1 promoter activities. An adenovirus expressing NF-kappaB inhibitor subunit-alpha inhibited NF-kappaB activities and also reversed the IL-1beta effect at the promoter activity and protein levels of ABCA1. In addition, IL-1beta could induce the production of reactive oxygen species (ROS), and N-acetyl-L-cysteine, a scavenger of ROS, reversed the decreased level of ABCA1 induced by IL-1beta. H(2)O(2) decreased ABCA1 at the mRNA and protein levels and the promoter activity. Thus our data provide strong evidence that ROS and NF-kappaB, but not LXR, mediate the IL-1beta-induced downregulation of ABCA1 via a novel transcriptional mechanism, which might play an important role of proinflammation in the alteration of lipid metabolism. PMID- 17135303 TI - Interleukin-15 responses to aging and unloading-induced skeletal muscle atrophy. AB - Interleukin-15 (IL-15) mRNA is constitutively expressed in skeletal muscle. Although IL-15 has proposed hypertrophic and anti-apoptotic roles in vitro, its role in skeletal muscle cells in vivo is less clear. The purpose of this study was to determine if skeletal muscle aging and unloading, two conditions known to promote muscle atrophy, would alter basal IL-15 expression in skeletal muscle. We hypothesized that IL-15 mRNA expression would increase as a result of both aging and muscle unloading and that muscle would express the mRNA for a functional trimeric IL-15 receptor (IL-15R). Two models of unloading were used in this study: hindlimb suspension (HS) in rats and wing unloading in quail. The absolute muscle wet weight of plantaris and soleus muscles from aged rats was significantly less when compared with muscles from young adult rats. Although 14 days of HS resulted in reduced muscle mass of plantaris and soleus muscles from young adult animals, this effect was not observed in muscles from aged animals. A significant aging times unloading interaction was observed for IL-15 mRNA in both rat soleus and plantaris muscles. Patagialis (PAT) muscles from aged quail retained a significant 12 and 6% of stretch-induced hypertrophy after 7 and 14 days of unloading, respectively. PAT muscles from young quail retained 15% hypertrophy at 7 days of unloading but regressed to control levels following 14 days of unloading. A main effect of age was observed on IL-15 mRNA expression in PAT muscles at 14 days of overload, 7 days of unloading, and 14 days of unloading. Skeletal muscle also expressed the mRNAs for a functional IL-15R composed of IL-15Ralpha, IL-2/15R-beta, and -gammac. Based on these data, we speculate that increases in IL-15 mRNA in response to atrophic stimuli may be an attempt to counteract muscle mass loss in skeletal muscles of old animals. Additional research is warranted to determine the importance of the IL-15/IL-15R system to counter muscle wasting. PMID- 17135304 TI - Twin pregnancy possibly associated with high semen quality. AB - BACKGROUND: Recent studies found an association between a long waiting time to pregnancy (TTP) and reduced probability of twinning and a reduced dizygotic (DZ) twinning rate in subfertile men. However, it remains unsolved whether semen quality is associated with twin offspring. We therefore studied the semen quality in a group of fathers of naturally conceived twins. METHODS: In this study, 37 fathers of DZ twins and 15 fathers of monozygotic (MZ) twins participated, and 349 normal fertile men served as a reference group. All men delivered a semen sample, underwent a physical examination and completed a questionnaire. RESULTS: After adjustment, fathers of DZ and MZ twins had 3.6 (95% CI 1.7; 5.4) and 4.6 (95% CI 2.0; 7.2) percentage points higher percentage of sperm cells with normal morphologic features and percentages of motile sperm cells were 11.5 (95% CI 7.2; 15.9) and 12.5 (95% CI 6.3; 18.6) percentage points higher than the reference group (P < 0.01). Fathers of DZ twins and MZ twins had 24.7 (95% CI; -9.1; 71.3) and 17.0% (-25.2%; 83.0%) higher sperm concentration than the reference group. CONCLUSIONS: Fathers of DZ twins had a better semen quality than the reference group, which supports the assumption that spontaneous DZ twinning rate can be used as a sensor of male fecundity of a population. PMID- 17135305 TI - Diastolic dysfunction is associated with poor survival in patients with cirrhosis with transjugular intrahepatic portosystemic shunt. AB - BACKGROUND: Transjugular intrahepatic portosystemic shunt (TIPS) is a treatment for portal hypertension-related complications. Accurate prediction of the outcome of patients treated with TIPS is important, because some patients have very short survival. Diastolic dysfunction is frequently observed in patients with cirrhosis. AIM: To investigate whether or not diastolic dysfunction can predict the outcome after TIPS. METHODS: Echocardiography with Doppler exploration was performed before and 28 days after TIPS insertion in 32 patients with cirrhosis. Several echocardiographic measures, including the early maximal ventricular filling velocity/late filling velocity (E/A) ratio as indicative of diastolic function, as well as laboratory, clinical and demographic variables were evaluated as predictors of survival. RESULTS: Univariate analysis revealed that the presence of diastolic dysfunction observed 28 days after TIPS (E/A ratio 1 survived. CONCLUSIONS: Diastolic dysfunction estimated using E/A ratio is a promising predictor of death in patients with cirrhosis who are treated with TIPS. PMID- 17135306 TI - Allelic deletion of the MEN1 gene in duodenal gastrin and somatostatin cell neoplasms and their precursor lesions. AB - BACKGROUND: Patients with a multiple endocrine neoplasia type 1 (MEN1)-associated Zollinger-Ellison syndrome (ZES) show multifocal duodenal gastrinomas and precursor lesions. AIMS: To test these lesions for loss of heterozygosity (LOH) of the MEN1 gene locus on chromosome 11q13, and to investigate whether the MEN1 related endocrine cell changes also involved somatostatin cells. MATERIAL AND METHODS: Tissue specimens from six patients with MEN1 and ZES were analysed by immunohistochemistry and immunofluorescence. LOH analysis was performed by fluorescence in situ hybridisation (FISH), using probes containing the MEN1 gene locus and the centromere 11 (C11) region. For simultaneous analysis of hormones and allelic deletions, a combined FISH/immunofluorescence protocol was established. RESULTS: 28 of a total of 33 duodenal neuroendocrine tumours (NETs) were gastrin-producing tumours; 13/28 (46.4%) revealed LOH on 11q13 and/or C11. Five of the NETs were somatostatin-expressing tumours, two revealing LOH. Allelic loss was detected in tumours as small as 300 microm (gastrin) and 400 microm (somatostatin) in diameter. The gastrin-producing tumours showed different deletion/retention patterns. Hyperplastic somatostatin cell lesions, similar to those of the gastrin cells, were present in all patients. The hyperplastic lesions of both cell lines consistently retained both 11q13 alleles. CONCLUSIONS: Allelic deletion of the MEN1 gene may reflect a pivotal event in the development of multifocal gastrin and somatostatin cell neoplasms in the duodenum of patients with MEN1. The observation of distinct deletion patterns in small synchronous tumours supports the concept that each gastrin-producing tumour in an individual MEN1 patient arises from an independent cell clone. PMID- 17135307 TI - Prevalence of oesophageal eosinophils and eosinophilic oesophagitis in adults: the population-based Kalixanda study. AB - BACKGROUND: Eosinophilic oesophagitis may be increasing but the prevalence in the general population remains unknown. Our aim was to assess this and the presence of eosinophils in the distal oesophageal epithelium in the community. METHODS: Oesophagogastroduodenoscopy was performed in a random sample (n = 1000) of the adult Swedish population (mean age 54 years, 49% men). Oesophageal biopsy samples were obtained from 2 cm above, and at, the Z-line. Any eosinophil infiltration of the epithelium was defined as "eosinophils present". Definite eosinophilic oesophagitis was defined as > or =20, probable as 15-19, and possible as 5-14 eosinophils/high-power field (HPF, at magnification x 40) in oesophageal biopsy specimens. RESULTS: Eosinophils were present in 48 subjects (4.8%, 95% CI 3.5 to 6.1%, mean age 54 years, 63% men), in 54% without troublesome reflux symptoms. Definite eosinophilic oesophagitis was present in four subjects (0.4%, 95% CI 0.01 to 0.8%, mean age 51 years, 75% men) and probable eosinophilic oesophagitis in seven subjects (0.7%, 95% CI 0.2 to 1.2%, mean age 58 years, 43% men). Erosive oesophagitis (OR = 2.99, 95% CI 1.58 to 5.66) and absence of dyspepsia (OR = 0.23, 95% CI 0.07 to 0.75) and Helicobacter pylori infection (OR = 0.41, 95% CI 0.19 to 0.92) were independent predictors for "eosinophils present". Definite eosinophilic oesophagitis was associated with dysphagia (2/66 vs 2/926, p = 0.025), and probable eosinophilic oesophagitis with narrowing of the oesophageal lumen (2/15 vs 5/978, p = 0.005). CONCLUSIONS: Oesophageal eosinophils were present in nearly 5% of the general population; approximately 1% had definite or probable eosinophilic oesophagitis. Oesophageal eosinophils may be a manifestation of reflux disease in adults, but the condition is as likely to be asymptomatic and go unrecognised. PMID- 17135308 TI - Heterozygous beta-globin gene mutations as a risk factor for iron accumulation and liver fibrosis in chronic hepatitis C. AB - BACKGROUND: Iron accumulation is a well-known risk factor for the progression of chronic hepatitis C (CHC) to fibrosis. However, the profibrogenic role of the genes controlling iron homeostasis is still controversial. AIM: To evaluate the relative role of haemachromatosis (HFE), ferroportin and beta-globin gene mutations in promoting iron accumulation and fibrosis in patients with CHC. METHODS: Genetic analysis was performed together with the assessment of hepatic iron content and histology in 100 consecutive HIV-antibody and hepatitis B surface antigen-negative patients with biopsy-proven CHC. RESULTS: Among the patients investigated, 12 were heterozygous for various beta-globin gene mutations (39[C-->T], IVS1.1[G-->A], 22 7 bp deletion and IVS1.6[T-->C]) and 29 carried HFE (C282Y, H63D and S65C) gene mutations. One further patient was heterozygous for both HFE (H63D) and beta-globin (39[C-->T]) variants, whereas 58 had the wild-type alleles of both the genes. Hepatic iron concentration (HIC) and hepatic stainable iron were significantly higher (p<0.05) in patients with CHC carrying beta-globin mutations than in those with HFE mutations or the wild-type alleles. Multivariate analysis confirmed that the presence of beta-globin mutations was independently associated with both HIC (p = 0.008) and hepatic stainable iron (odds ratio (OR) 6.11; 95% CI 1.56 to 23.92; p = 0.009). Moderate/severe fibrosis or cirrhosis (Ishak's score >2) was observed in 48 of 100 patients. Logistic regression demonstrated that age (OR 1.05; 95% CI 1.02 to 1.09; p<0.005) and beta-globin mutations (OR 4.99; 95% CI 1.22 to 20.3; p = 0.025) were independent predictors of the severity of fibrosis. CONCLUSIONS: Heterozygosis for beta-globin mutations is a novel risk factor for both hepatic iron accumulation and the progression to fibrosis in patients with CHC. PMID- 17135309 TI - The epidemiology of microscopic colitis: a population based study in Olmsted County, Minnesota. AB - OBJECTIVE: Although the epidemiology of microscopic colitis has been described in Europe, no such data exist from North America. We studied the incidence, prevalence and temporal trends of microscopic colitis in a geographically defined US population. DESIGN AND SETTING: In this population based cohort study, residents of Olmsted County, Minnesota, with a new diagnosis of microscopic colitis, and all who had colon biopsies for evaluation of diarrhoea, between 1 January 1985 and 31 December 2001 were identified. Biopsies were reviewed for confirmation (cases) and to identify missed cases (diarrhoea biopsies). MAIN OUTCOME MEASURES: Incidence rates, age and sex adjusted to the 2000 US white population. Poisson regression assessed the association of calendar period, age and sex with incidence. RESULTS: We identified 130 incident cases for an overall rate of 8.6 cases per 100,000 person-years. There was a significant secular trend, with incidence increasing from 1.1 per 100,000 early in the study to 19.6 per 100,000 by the end (p<0.001). Rates increased with age (p<0.001). By subtype, the incidence was 3.1 per 100,000 for collagenous colitis and 5.5 per 100,000 for lymphocytic colitis. Collagenous colitis was associated with female sex (p<0.001) but lymphocytic colitis was not. Prevalence (per 100,000 persons) on 31 December 2001 was 103.0 (39.3 for collagenous colitis and 63.7 for lymphocytic colitis). CONCLUSIONS: The incidence of microscopic colitis has increased significantly over time, and by the end of the study, the incidence and prevalence were significantly higher than reported previously. Microscopic colitis is associated with older age, and collagenous colitis is associated with female sex. PMID- 17135310 TI - Endoscopic mucosal resection for flat neoplasia in chronic ulcerative colitis: can we change the endoscopic management paradigm? AB - BACKGROUND: The potential of endoscopic mucosal resection (EMR) for treating flat dysplastic lesions in chronic ulcerative colitis (CUC) has not been addressed so far. Historically, such lesions were referred for colectomy. Furthermore, there are only limited data to support endoscopic resection of exophytic adenoma-like mass (ALM) lesions in colitis. AIMS: To evaluate the safety and clinical outcomes of patients with colitis undergoing EMR for Paris class 0-II and class I ALM compared with sporadic controls. Secondary aims were to re-evaluate the prevalence, anatomical "mapping" and histopathological characteristics of both Paris class 0-II and class I lesions in the context of CUC. METHODS: Prospective clinical, pathological and outcome data of patients with colitis-associated Paris class 0-II and Paris class I ALM treated with EMR (primary end points being colorectal cancer development, resection efficacy, metachronous lesion rates and post-resection recurrence rates) were compared with those of sporadic controls. RESULTS: 204 lesions were diagnosed in 169 patients during the study period: 167 (82%) diagnosed at "entry" colonoscopy, and 36 (18%) diagnosed at follow-up. 170 ALMs, 18 dysplasia-associated lesion masses (DALMs) and 16 cancers were diagnosed. A total of 4316 colonoscopies were performed throughout the study period (median per patient: 6; range: 1-8). The median follow-up period for the complete cohort was 4.1 years (range: 3.6-5.21). 1675 controls were included from our prospective database of patients without CUC who had undergone EMR for sporadic Paris class 0-II and snare polypectomy of Paris type I lesions from 1998 onwards, and were considered to be at moderate to high lifetime risk of colorectal cancer. 3792 colonoscopies were performed throughout the study period in this group (median per patient: 4; range: 1-7). The median follow-up period was 4.8 years (range: 2.9-5.2). No statistically significant differences were observed between the CUC study group and controls with respect to age, sex, median number of colonoscopies per patient, median follow-up duration, post resection complications, median lesional diameter or interval cancer rates. However, there was a significant between-group difference regarding the prevalence of Paris class 0-II lesions in the CUC group (82/155 (61%)) compared with controls (285/801 (35%); chi(2) = 31.13; p<0.001). Furthermore, recurrence rates of lateral spreading tumours were higher in the colitis cohort (1/7 (14%)) than among controls (0/10 (0%); p = 0.048 (95% CI 11.64% to 40.21%)). CONCLUSIONS: Flat DALM, similarly to Paris class I ALM, can be managed safely by EMR in CUC. A change in management paradigm to include EMR for the resection of flat dysplastic lesions in selected cases is proposed. PMID- 17135311 TI - Suppression of transforming growth factor beta signalling aborts caerulein induced pancreatitis and eliminates restricted stimulation at high caerulein concentrations. AB - BACKGROUND: Transforming growth factors betas (TGF-betas) are implicated in pancreatic tissue repair but their role in acute pancreatitis is not known. To determine whether endogenous TGF-betas modulate the course of caerulein induced acute pancreatitis, caerulein was administered to wild-type (FVB-/-) and transgenic mice that are heterozygous (FVB+/-) for expression of a dominant negative type II TGF-beta receptor. METHODS: After 7 hourly supramaximal injections of caerulein, the pancreas was evaluated histologically and serum was assayed for amylase and lipase levels. Next, the effects of caerulein on amylase secretion were determined in mouse pancreatic acini, and cholecystokinin (CCK) receptor expression was assessed. RESULTS: The normal mouse pancreas was devoid of inflammatory cells whereas the pancreas from transgenic mice contained lymphocytic infiltrates. Caerulein injection in wild-type mice resulted in 6- and 36-fold increases in serum amylase and lipase levels, respectively, increased serum trypsinogen activation peptide (TAP) levels, gross oedema and a marked inflammatory response in the pancreas that consisted mainly of neutrophils and macrophages. By contrast, FVB+/- mice exhibited minimal alterations in response to caerulein with attenuated neutrophil-macrophage infiltrates. Moreover, acini from FVB+/- mice did not exhibit restricted stimulation at high caerulein concentrations, even though CCK receptor mRNA levels were not decreased. CONCLUSION: Our findings indicate that a functional TGF-beta signalling pathway may be required for caerulein to induce acute pancreatitis and for the CCK receptor to induce acinar cell damage at high ligand concentrations. Our results also support the concept that restricted stimulation at high caerulein concentrations contributes to the ability of caerulein to induce acute pancreatitis. PMID- 17135312 TI - OGG1 and MYH are involved in the incision of trivalent arsenical-induced DNA adducts. AB - Since trivalent arsenicals are known to induce oxidative DNA damage in human cells, we asked if they induce other types of DNA damage and how these DNA damages are repaired. Treatment of human promyelocytic leukemia NB4 cells with 0.5 microM As2O3 for 30 min induced no DNA breaks, as analyzed by a standard comet assay. However, breaks were detected if these cells were then digested with endonuclease III (EnIII), formamidopyrimidine-DNA glycosylase (Fpg), or a nuclear extract (NE) of NB4 cells. Using either H2O2-Fe treated nuclei or As2O3-treated cells, digestion with either NE or EnIII+Fpg generated the same amount of breaks, and subsequent treatment with EnIII+Fpg resulted in no increase in breaks in NE digested cells and vice versa. The human cell lines, defective in nucleotide excision protein, such as XPA, XPD, and XPG, excised UVC-induced adducts less rapidly than normal fibroblasts, but excised As2O3-adducts at the same rate as the normal cells. Immunodepletion of the NE with antibody against OGG1 or MYH decreased the incision of DNA adducts induced by As2O3, NaAsO2, monomethylarsonic acid, and dimethylarsinic acid, while antibodies against XPA, XPB, XPD, XPF, or XPG, did not. These results suggest that these trivalent arsenicals induce the formation of only oxidative DNA adducts and that OGG1 and MYH are involved in these incision processes. PMID- 17135313 TI - Genome sequence, full-length infectious cDNA clone, and mapping of viral double stranded RNA accumulation determinant of hypovirus CHV1-EP721. AB - Cryphonectria parasitica strain EP721 is infected with a strain of hypovirus CHV1, CHV1-EP721, and exhibits typical hypovirulence-associated traits such as reduced pigmentation and reduced asexual sporulation. However, the accumulation of the viral double-stranded RNA (dsRNA) in this hypovirus-infected C. parasitica strain is atypically low. We now report the complete nucleotide sequence and construction of a full-length infectious cDNA clone for hypovirus CHV1-EP721. The genome sequence of CHV1-EP721 was determined to be 12,724 bp in length and to share extensive homology with two other hypovirus strains, CHV1-Euro7 and CHV1 EP713, with an average of 99% and 90% identities at the nucleotide level and 99% and 92% identities at the amino acid level, respectively. CHV1-EP721 was successfully introduced into virus-free fungal host strain EP721(-v) by transfection with transcripts derived from a full-length viral cDNA. The transfected strain had a phenotype indistinguishable from that of EP721, and the accumulation of CHV1-EP721 dsRNA in the transfectant was lower than those transfected by CHV1-Euro7 and CHV1-EP713 transcripts. Through the construction of chimeric viruses by domain swapping using infectious cDNA clones of CHV1-EP721, CHV1-EP713, and CHV1-Euro7 hypoviruses, the determinant for the low level of viral dsRNA accumulation in CHV1-EP721 was mapped to the second of two CHV1-EP721 open reading frames (ORFs), ORF B. Further refined swapping of domains within ORF B identified a 2.5-kb coding region between p48 and the polymerase domain of CHV1 EP721 as being responsible for the low viral dsRNA accumulation. Evidence is also provided that low rates of hypovirus transmission through conidial spores correlates with low viral dsRNA accumulation. PMID- 17135314 TI - The human TRIM5alpha restriction factor mediates accelerated uncoating of the N tropic murine leukemia virus capsid. AB - The host cell factors TRIM5alpha(hu) and Fv-1 restrict N-tropic murine leukemia virus (N-MLV) infection at an early postentry step before or after reverse transcription, respectively. Interestingly, the identity of residue 110 of the MLV capsid determines susceptibility to both TRIM5alpha(hu) and Fv-1. In this study, we investigate the fate of the MLV capsid in cells expressing either the TRIM5alpha(hu) or Fv-1 restriction factor. The expression of TRIM5alpha(hu), but not Fv-1, specifically promoted the premature conversion of particulate N-MLV capsids within infected cells to soluble capsid proteins. The TRIM5alpha(hu) mediated disassembly of particulate N-MLV capsids was dependent upon residue 110 of the viral capsid. Furthermore, the deletion or disruption of TRIM5alpha(hu) domains necessary for potent N-MLV restriction completely abrogated the disappearance of particulate N-MLV capsids observed with wild-type TRIM5alpha(hu). These results suggest that premature disassembly of the viral capsid contributes to the restriction of N-MLV infection by TRIM5alpha(hu), but not by Fv-1. PMID- 17135315 TI - Mitotic kinesin-like protein 2 binds and colocalizes with papillomavirus E2 during mitosis. AB - MKlp2 is a kinesin-like motor protein of the central mitotic spindle required for completion of cytokinesis. Papillomavirus E2 is a sequence specific DNA binding protein that regulates viral transcription and replication and is responsible for partitioning viral episomes into daughter cells during cell division. We demonstrate that MKlp2 specifically associates with the E2 protein during mitosis. Using chromatin immunoprecipitation, we show viral genomes are in complex with MKlp2 only within this stage of cell cycle. By immunofluorescence, a subpopulation of papillomavirus E2 colocalizes with MKlp2 in the midbody/midplate during late mitosis. We conclude that during specific stages of mitosis, the papillomavirus E2 protein binds to MKlp2, and infer that association with this motor protein ensures viral genome partitioning during cytokinesis. PMID- 17135316 TI - Tomato chlorotic mottle virus is a target of RNA silencing but the presence of specific short interfering RNAs does not guarantee resistance in transgenic plants. AB - Tomato chlorotic mottle virus (ToCMoV) is a begomovirus found widespread in tomato fields in Brazil. ToCMoV isolate BA-Se1 (ToCMoV-[BA-Se1]) was shown to trigger the plant RNA silencing surveillance in different host plants and, coinciding with a decrease in viral DNA levels, small interfering RNAs (siRNAs) specific to ToCMoV-[BA-Se1] accumulated in infected plants. Although not homogeneously distributed, the siRNA population in both infected Nicotiana benthamiana and tomato plants represented the entire DNA-A and DNA-B genomes. We determined that in N. benthamiana, the primary targets corresponded to the 5' end of AC1 and the embedded AC4, the intergenic region and 5' end of AV1 and overlapping central part of AC5. Subsequently, transgenic N. benthamiana plants were generated that were preprogrammed to express double-stranded RNA corresponding to this most targeted portion of the virus genome by using an intron-hairpin construct. These plants were shown to indeed produce ToCMoV specific siRNAs. When challenge inoculated, most transgenic lines showed significant delays in symptom development, and two lines had immune plants. Interestingly, the levels of transgene-produced siRNAs were similar in resistant and susceptible siblings of the same line. This indicates that, in contrast to RNA viruses, the mere presence of transgene siRNAs corresponding to DNA virus sequences does not guarantee virus resistance and that other factors may play a role in determining RNA-mediated resistance to DNA viruses. PMID- 17135317 TI - Evidence for the existence of the loop E motif of Potato spindle tuber viroid in vivo. AB - RNA motifs comprising nucleotides that interact through non-Watson-Crick base pairing play critical roles in RNA functions, often by serving as the sites for RNA-RNA, RNA-protein, or RNA small ligand interactions. The structures of viral and viroid RNA motifs are studied commonly by in vitro, computational, and mutagenesis approaches. Demonstration of the in vivo existence of a motif will help establish its biological significance and promote mechanistic studies on its functions. By using UV cross-linking and primer extension, we have obtained direct evidence for the in vivo existence of the loop E motif of Potato spindle tuber viroid. We present our findings and discuss their biological implications. PMID- 17135318 TI - Reassortment and concerted evolution in banana bunchy top virus genomes. AB - The nanovirus Banana bunchy top virus (BBTV) has six standard components in its genome and occasionally contains components encoding additional Rep (replication initiation protein) genes. Phylogenetic network analysis of coding sequences of DNA 1 and 3 confirmed the two major groups of BBTV, a Pacific and an Asian group, but show evidence of web-like phylogenies for some genes. Phylogenetic analysis of 102 major common regions (CR-Ms) from all six components showed a possible concerted evolution within the Pacific group, which is likely due to recombination in this region. The CR-M of additional Rep genes is close to that of DNA 1 and 2. Comparison of tree topologies constructed with DNA 1 and DNA 3 coding sequences of 14 BBTV isolates showed distinct phylogenetic histories based on Kishino-Hasegawa and Shimodaira-Hasegawa tests. The results of principal component analysis of amino acid and codon usages indicate that DNA 1 and 3 have a codon bias different from that of all other genes of nanoviruses, including all currently known additional Rep genes of BBTV, which suggests a possible ancient genome reassortment event between distinctive nanoviruses. PMID- 17135319 TI - Regulation of hepadnavirus reverse transcription by dynamic nucleocapsid phosphorylation. AB - Reverse transcription, an essential step in the life cycle of all retroelements, is a complex, multistep process whose regulation is not yet clearly understood. We have recently shown that reverse transcription in the pararetrovirus duck hepatitis B virus is associated with complete dephosphorylation of the viral core protein, which forms the nucleocapsid wherein reverse transcription takes place. Here we present a genetic study of the role of this dynamic nucleocapsid phosphorylation in regulating viral reverse transcription. Detailed analyses of the reverse transcription products synthesized within nucleocapsids composed of core phosphorylation site mutants revealed that alanine substitutions, mimicking the nonphosphorylated state, completely blocked reverse transcription at a very early stage. In contrast, aspartate substitutions, mimicking the phosphorylated state, allowed complete first-strand DNA synthesis but were severely defective in accumulating mature double-stranded DNA. The latter defect was due to a combination of mutant nucleocapsid instability during maturation and a block in mature second-strand DNA synthesis. Thus, the reversible phosphorylation of the nucleocapsids regulates the ordered progression of reverse transcription. PMID- 17135320 TI - The transdominant endogenous retrovirus enJS56A1 associates with and blocks intracellular trafficking of Jaagsiekte sheep retrovirus Gag. AB - The sheep genome harbors approximately 20 endogenous retroviruses (enJSRVs) highly related to the exogenous Jaagsiekte sheep retrovirus (JSRV). One of the enJSRV loci, enJS56A1, acts as a unique restriction factor by blocking JSRV in a transdominant fashion at a late stage of the retroviral cycle. To better understand the molecular basis of this restriction (termed JLR, for JSRV late restriction), we functionally characterized JSRV and enJS56A1 Gag proteins. We identified the putative JSRV Gag membrane binding and late domains and determined their lack of involvement in JLR. In addition, by using enJS56A1 truncation mutants, we established that the entire Gag protein is necessary to restrict JSRV exit. By using differentially tagged viruses, we observed, by confocal microscopy, colocalization between JSRV and enJS56A1 Gag proteins. By coimmunoprecipitation and molecular complementation analyses, we also revealed intracellular association and likely coassembly between JSRV and enJS56A1 Gag proteins. Interestingly, JSRV and enJS56A1 Gag proteins showed distinct intracellular targeting: JSRV exhibited pericentrosomal accumulation of Gag staining, while enJS56A1 Gag did not accumulate in this region. Furthermore, the number of cells displaying pericentrosomal JSRV Gag was drastically reduced in the presence of enJS56A1. We identified amino acid residue R21 in JSRV Gag as the primary determinant of centrosome targeting. We concluded that JLR is dependent on a Gag-Gag interaction between enJS56A1 and JSRV leading to altered cellular localization of the latter. PMID- 17135321 TI - Long-lasting decrease in viremia in macaques chronically infected with simian immunodeficiency virus SIVmac251 after therapeutic DNA immunization. AB - Rhesus macaques chronically infected with highly pathogenic simian immunodeficiency virus (SIV) SIVmac251 were treated with antiretroviral drugs and vaccinated with combinations of DNA vectors expressing SIV antigens. Vaccination during therapy increased cellular immune responses. After the animals were released from therapy, the virus levels of 12 immunized animals were significantly lower (P = 0.001) compared to those of 11 animals treated with only antiretroviral drugs. Vaccinated animals showed a persistent increase in immune responses, thus indicating both a virological and an immunological benefit following DNA therapeutic vaccination. Several animals show a long-lasting decrease in viremia, suggesting that therapeutic vaccination may provide an additional benefit to antiretroviral therapy. PMID- 17135322 TI - Heterogeneity of Raft-type membrane microdomains associated with VP4, the rotavirus spike protein, in Caco-2 and MA 104 cells. AB - Previous studies have shown that rotavirus virions, a major cause of infantile diarrhea, assemble within small intestinal enterocytes and are released at the apical pole without significant cell lysis. In contrast, for the poorly differentiated kidney epithelial MA 104 cells, which have been used extensively to study rotavirus assembly, it has been shown that rotavirus is released by cell lysis. The subsequent discovery that rotavirus particles associate with raft-type membrane microdomains (RTM) in Caco-2 cells provided a simple explanation for rotavirus polarized targeting. However, the results presented here, together with those recently published by another group, demonstrate that rotavirus also associates with RTM in MA 104 cells, thus indicating that a simple interaction of rotavirus with rafts is not sufficient to explain its apical targeting in intestinal cells. In the present study, we explore the possibility that RTM may have distinct physicochemical properties that may account for the differences observed in the rotavirus cell cycle between MA 104 and Caco-2 cells. We show here that VP4 association with rafts is sensitive to cholesterol extraction by methyl-beta-cyclodextrin treatment in MA 104 cells and insensitive in Caco-2 cells. Using the VP4 spike protein as bait, VP4-enriched raft subsets were immunopurified. They contained 10 to 15% of the lipids present in total raft membranes. We found that the nature and proportion of phospholipids and glycosphingolipids were different between the two cell lines. We propose that this raft heterogeneity may support the cell type dependency of virus assembly and release. PMID- 17135323 TI - A novel repeat-associated small interfering RNA-mediated silencing pathway downregulates complementary sense gypsy transcripts in somatic cells of the Drosophila ovary. AB - Replication of the gypsy endogenous retrovirus involves contamination of the female germ line by adjacent somatic tissues. This is prevented by flam, an as yet-uncloned heterochromatic pericentromeric locus, at the level of transcript accumulation in these somatic ovarian tissues. We tested the effect of a presumptive RNA silencing mechanism on the accumulation of RNAs produced by constructs containing various gypsy sequences and report that the efficiency of silencing is indeed correlated with the amount of complementary RNAs, 25 to 30 nucleotides in length, in the ovary. For instance, while these RNAs were found to display a three- to fivefold excess of the antisense strands, only the transcripts that contain the complementary sense gypsy sequences could be repressed, indicating that they are targeted at the RNA, not DNA, level. Their size and asymmetry in strand polarity are typical of the novel repeat-associated small interfering RNA (rasiRNA)-mediated pathway, recently suspected to prevent the deleterious expression of selfish DNA specifically in the germ line. Unlike microRNAs (but like rasiRNAs and, surprisingly, siRNAs as well), gypsy rasiRNAs are modified at the 3' end. The rasiRNA-associated protein Piwi (but not Aub) is required for gypsy silencing, whereas Dicer-2 (which makes siRNAs) is not. In contrast, piwi, aub, and flam do not appear to affect somatic siRNA-mediated silencing. The amount of gypsy rasiRNAs is genetically determined by the flam locus in a provirus copy number-independent manner and is triggered in the somatic tissues by some pericentromeric provirus(es), which are thereby able to protect the germ line from retroviral invasion. PMID- 17135324 TI - Not all cytokine-producing CD8+ T cells suppress simian immunodeficiency virus replication. AB - Current assays of CD8+ T-lymphocyte function measure cytokine production rather than the ability of these lymphocytes to suppress viral replication. Here we show that CD8+ T-cell clones recognizing the same epitope vary enormously in the ability to suppress simian immunodeficiency virus SIVmac239 replication in an in vitro suppression assay. However, all Nef(165-173)IW9- and Vif(66-73)HW8-specific clones from elite controllers effectively suppressed SIV replication. Interestingly, in vitro suppression efficacy was not always associated with the ability to produce gamma interferon, tumor necrosis factor alpha, or interleukin 2. PMID- 17135325 TI - Measles virus infection of SLAM (CD150) knockin mice reproduces tropism and immunosuppression in human infection. AB - The human signaling lymphocyte activation molecule (SLAM, also called CD150), a regulator of antigen-driven T-cell responses and macrophage functions, acts as a cellular receptor for measles virus (MV), and its V domain is necessary and sufficient for receptor function. We report here the generation of SLAM knockin mice in which the V domain of mouse SLAM was replaced by that of human SLAM. The chimeric SLAM had an expected distribution and normal function in the knockin mice. Splenocytes from the SLAM knockin mice permitted the in vitro growth of a virulent MV strain but not that of the Edmonston vaccine strain. Unlike in vitro infection, MV could grow only in SLAM knockin mice that also lacked the type I interferon receptor (IFNAR). After intraperitoneal or intranasal inoculation, MV was detected in the spleen and lymph nodes throughout the body but not in the thymus. Notably, the virus appeared first in the mediastinal lymph node after intranasal inoculation. Splenocytes from MV-infected IFNAR(-/-) SLAM knockin mice showed suppression of proliferative responses to concanavalin A. Thus, MV infection of SLAM knockin mice reproduces lymphotropism and immunosuppression in human infection, serving as a useful small animal model for measles. PMID- 17135326 TI - Involvement of the PA28gamma-dependent pathway in insulin resistance induced by hepatitis C virus core protein. AB - The hepatitis C virus (HCV) core protein is a component of nucleocapsids and a pathogenic factor for hepatitis C. Several epidemiological and experimental studies have suggested that HCV infection is associated with insulin resistance, leading to type 2 diabetes. We have previously reported that HCV core gene transgenic (PA28gamma(+/+)CoreTg) mice develop marked insulin resistance and that the HCV core protein is degraded in the nucleus through a PA28gamma-dependent pathway. In this study, we examined whether PA28gamma is required for HCV core induced insulin resistance in vivo. HCV core gene-transgenic mice lacking the PA28gamma gene (PA28gamma(-/-)CoreTg) were prepared by mating of PA28gamma(+/+)CoreTg with PA28gamma-knockout mice. Although there was no significant difference in the glucose tolerance test results among the mice, the insulin sensitivity in PA28gamma(-/-)CoreTg mice was recovered to a normal level in the insulin tolerance test. Tyrosine phosphorylation of insulin receptor substrate 1 (IRS1), production of IRS2, and phosphorylation of Akt were suppressed in the livers of PA28gamma(+/+)CoreTg mice in response to insulin stimulation, whereas they were restored in the livers of PA28gamma(-/-)CoreTg mice. Furthermore, activation of the tumor necrosis factor alpha promoter in human liver cell lines or mice by the HCV core protein was suppressed by the knockdown or knockout of the PA28gamma gene. These results suggest that the HCV core protein suppresses insulin signaling through a PA28gamma-dependent pathway. PMID- 17135327 TI - From innovation to practice: initiation, implementation and evaluation of a physician-based physical activity promotion programme in Finland. AB - In 2001, a collaborative Physical Activity Prescription Programme (PAPP) was started in Finland to increase physical activity (PA) counselling among physicians, especially in primary care. This article describes the initiation, implementation and evaluation of PAPP. Five actions were implemented to reach the programme goal: (i) developing a counselling approach for physicians; (ii) providing easy and open access to counselling material; (iii) facilitating physicians' uptake and adoption of the counselling approach; (iv) disseminating information about the counselling approach to physicians, health and exercise professionals and decision-makers and (v) raising financial resources to cover programme expenses. Evaluation was based on the dimensions of the RE-AIM framework: reach, effectiveness, adoption, implementation and maintenance. Effectiveness and adoption were evaluated with two questions added to the annual survey of the Finnish Medical Association to all practising physicians in the year 2002 (n = 16 692) and 2004 (n = 17 170). The 4-year PAPP was successful in reaching health care units (Reach), accomplishing most of the implementation actions (Implementation) and initiating local projects for institutionalizing the prescription-based counselling approach, 'Prex' (Maintenance). However, at the national level, the programme was not effective in increasing the frequency of asking about patients' PA habits (Effectiveness) or the frequency of using 'Prex' or other written material in PA counselling among physicians (Adoption). To improve the latter two, the duration of the programme would have had to be extended with more effort at strengthening physicians' confidence in PA counselling and knowledge about its effectiveness. Also, a more systematic approach would have been necessary to facilitate inter-sectoral network for adopting 'Prex' as a counselling tool at the local level. PMID- 17135328 TI - Randomised controlled trial on whether advance knowledge of prostate-specific antigen testing improves participant reporting of unprotected sex. AB - OBJECTIVES: To determine whether the process of informing research participants that they would be tested for the presence of a biological marker of semen exposure would reduce bias in their reports of unprotected sex. METHODS: A randomised trial of 210 female sex workers from Mombasa, Kenya, was conducted, where half the group had advance knowledge (via the request for informed consent) that they would be tested for prostate-specific antigen (PSA) in their vaginal fluid before they reported on sex and condom use for the past 48 h. The other half were invited to participate (via additional informed consent) in the test for PSA after they had already consented to be questioned and reported on these sexual behaviours. A trained nurse instructed participants to self-swab to collect vaginal fluid specimens, which were tested for PSA using ELISA. RESULTS: Reporting of unprotected sex did not differ between those with advance knowledge of the test for PSA and those without this knowledge (14.3% v 11.4%, respectively; p = 0.27). Surprisingly, more women with advance knowledge (15.8%) had discrepant self reports and PSA results than women without advance knowledge (9.1%); however, the difference was not statistically significant (OR 1.9; 95% CI 0.8 to 4.5). CONCLUSIONS: Knowing that one's answers to a questionnaire could be verified with a biological marker of semen exposure did not make respondents more likely to report unprotected sex. PMID- 17135329 TI - Excellence in sexually transmitted infection (STI) diagnostics: recognition of past successes and strategies for the future. AB - Diagnostic advances do not generally receive the recognition given to prevention and treatment contributions, for the control and management of infectious diseases including sexually transmitted infections (STIs). In order to identify seminal diagnostic contributions over a half century (1950-2000), the Editorial Board of the WHO Sexually Transmitted Diseases Diagnostics Initiative (SDI) Publication Review or "electronic journal club" were asked to nominate their choices of peer-reviewed publications for special recognition. From 43 nominations, 13 were voted by a panel of 25 "experts" as having made the most significant contributions. The 1964 article by Thayer and Martin, which identified a selective media for gonococcal culture, was chosen unanimously by all panel members and is identified as the classic STI diagnostic article for this era. PMID- 17135330 TI - Men who have sex with men in Great Britain: comparison of a self-selected internet sample with a national probability sample. AB - OBJECTIVES: To compare the characteristics of a self-selected, convenience sample of men who have sex with men (MSM) recruited through the internet with MSM drawn from a national probability survey in Great Britain. METHODS: The internet sample (n = 2065) was recruited through two popular websites for homosexual men in Great Britain in May and June 2003. This sample was compared with MSM (n = 117) from the National Survey of Sexual Attitudes and Lifestyles (Natsal), a probability sample survey of adults resident in Great Britain conducted between May 1999 and February 2001. RESULTS: No significant differences were observed between the samples on a range of sociodemographic and behavioural variables (p>0.05). However, men from the internet sample were younger (p<0.001) and more likely to be students (p = 0.001), but less likely to live in London (p = 0.001) or report good health (p = 0.014). Although both samples were equally likely to report testing for HIV, men from the internet sample were more likely to report a sexually transmitted infection in the past year (16.9% v 4.8%, adjusted odds ratio 4.14, 95% CI 1.76 to 9.74; p = 0.001), anal intercourse (76.9% v 63.3%; p = 0.001) and unprotected anal intercourse in the past 3 months (45% v 36.6%; p = 0.064). CONCLUSIONS: The internet provides a means of recruiting a self-selected, convenience sample of MSM whose social and demographic characteristics are broadly similar to those of MSM drawn from a national probability survey. However, estimates of high-risk sexual behaviour based on internet convenience samples are likely to overestimate levels of sexual risk behaviour in the wider MSM population. PMID- 17135331 TI - Sequence diversity at the proximal 14q32.1 SERPIN subcluster: evidence for natural selection favoring the pseudogenization of SERPINA2. AB - The superfamily of serine protease inhibitors (SERPINs) plays a key role in controlling the activity of proteinases in diverse biological processes. alpha1 antitrypsin (SERPINA1), the most studied member of this family, is encoded by a gene located within the proximal 14q32.1 SERPIN subcluster, together with the highly homologous alpha1-antitrypsin-like sequence (SERPINA2), which was previously proposed to be a pseudogene. Here, we performed a resequencing study encompassing both SERPINA1 and SERPINA2 as well as the adjacent gene coding for corticosteroid-binding globulin (SERPINA6) in samples from Europe and West Africa. In the African sample, we found that a common haplotype carrying a 2-kb deletion in the SERPINA2 gene is associated with remarkable long-range homozygozity as if it was quickly driven to high frequency by natural selection acting on an advantageous variant. An analysis of the HapMap Phase I data for the Yoruba sample confirmed that variation in this subcluster carries a strong signal of positive selection. We also show that the SERPINA2 gene is expressed and probably encodes a functional SERPIN. Finally, comparisons with orthologous sequences in nonhuman primates showed that SERPINA2 is present in some great apes, but in chimpanzees it was lost by a deletion event independent from that observed in humans. In agreement with the "less is more" hypothesis, we propose that loss of SERPINA2 is an ongoing process associated with a selective advantage during recent primate evolution, possibly because of a role in fertility or in host-pathogen interactions. PMID- 17135332 TI - The evolution of GABAA receptor-like genes. AB - The inhibitory ligand-gated ion channel family of receptors, including the type A gamma-aminobutryic acid (GABA(A)) and glycine receptors, mediates inhibitory neurotransmissions in the central nervous system. In this study, GABA receptor (GABR) evolution was explored through comparative genomics using genomes that span divergent lineages. GABA(A)/Gly receptor-like (GRL) gene sequences were retrieved from the genomes of various species ranging from mammal to fish to worm and subjected to cross-species comparison. All vertebrate GRL gene sets in the study but no invertebrate ones exhibit the extensive and conserved pattern of gene clustering that is characteristic of human GABR genes, indicating that the gene clusters were established early in vertebrate evolution, after divergence from the invertebrates. Moreover, the vertebrate gene structure is highly conserved with a basic 9-coding exon structure, whereas, as well as being diverse in copy numbers and chromosomal loci, the invertebrate GRL genes display a variety of gene structures. Remarkably, the invertebrates each possess a unique GRL gene pair that lies in neighboring loci within their respective genomes: zc482.5 and zc482.1 in roundworm, CG8916 and CG17336 in fruitfly, Ci4249 and Ci4254 in Ciona, and these were revealed by phylogenetic analysis to be homologous to human GABR alpha and beta subunits, respectively. The phylogenetic classification of these genes is also corroborated by experimental ligand-binding measurements using recombinant gene products. Furthermore, the 3 invertebrate gene pairs harbor characteristic key residues and exhibit similarities in intron positions to their vertebrate counterparts. The results strongly indicate that such a gene pair originally existed in the bilaterian ancestor from which all 3 phyla evolved and suggest that the extant GABR clusters arose from an ancestral alpha-beta subunit gene pair gave rise to the extant GABR clusters. PMID- 17135333 TI - Functional diversification of B MADS-box homeotic regulators of flower development: Adaptive evolution in protein-protein interaction domains after major gene duplication events. AB - B-class MADS-box genes have been shown to be the key regulators of petal and stamen specification in several eudicot model species such as Arabidopsis thaliana, Antirrhinum majus, and Petunia hybrida. Orthologs of these genes have been found across angiosperms and gymnosperms, and it is thought that the basic regulatory function of B proteins is conserved in seed plant lineages. The evolution of B genes is characterized by numerous duplications that might represent key elements fostering the functional diversification of duplicates with a deep impact on their role in the evolution of the floral developmental program. To evaluate this, we performed a rigorous statistical analysis with B gene sequences. Using maximum likelihood and Bayesian methods, we estimated molecular substitution rates and determined the selective regimes operating at each residue of B proteins. We implemented tests that rely on phylogenetic hypotheses and codon substitution models to detect significant differences in substitution rates (DSRs) and sites under positive adaptive selection (PS) in specific lineages before and after duplication events. With these methods, we identified several protein residues fixed by PS shortly after the origin of PISTILLATA-like and APETALA3-like lineages in angiosperms and shortly after the origin of the euAP3-like lineage in core eudicots, the 2 main B gene duplications. The residues inferred to have been fixed by positive selection lie mostly within the K domain of the protein, which is key to promote heterodimerization. Additionally, we used a likelihood method that accommodates DSRs among lineages to estimate duplication dates for AP3-PI and euAP3-TM6, calibrating with data from the fossil record. The dates obtained are consistent with angiosperm origins and diversification of core eudicots. Our results strongly suggest that novel multimer formation with other MADS proteins could have been crucial for the functional divergence of B MADS-box genes. We thus propose a mechanism of functional diversification and persistence of gene duplicates by the appearance of novel multimerization capabilities after duplications. Multimer formation in different combinations of regulatory proteins can be a mechanistic basis for the origin of novel regulatory functions and a gene regulatory mechanism for the appearance of morphological innovations. PMID- 17135334 TI - Mechanisms and rates of birth and death of dispersed duplicated genes during the evolution of a multigene family in diploid and tetraploid wheats. AB - A family of 5 genes that evolved within the past 1.9 Myr in diploid wheat was characterized. The ancestral gene, ALP-A1, is on chromosome 1A and encodes an aci reductone dioxygenase-like protein. The duplicated genes ALP-A2, ALP-A3, ALP A4.1, and ALP-A4.2 acquired complete coding sequences but lost the original promoter. They are on chromosomes 4A, 2A, 6A and 6A, respectively, and evolved sequentially, the youngest duplicated gene always producing the next duplicate. It is shown that dispersed gene duplication rate consists of the primary rate (duplications of ancestral genes) and the secondary rate (duplications of genes that had been generated by recent duplications). The primary rate was 2.5 x 10( 3) gene(-1) Myr(-1) in diploid wheat. The secondary rate was 5.2 x 10(-2) gene( 1) Myr(-1) in the ALP family. The 20-fold acceleration of the secondary rate was caused by the insertion of the ALP-A2 gene into a novel type transposon. Only the ALP-A1 and ALP-A3 genes are transcribed. The transcription of ALP-A3 is directed by a promoter within a DNA fragment similar to a CACTA type of DNA transposons, making ALP-A3 a new gene. The ALP-A3 transcript is longer than that of the ALP A1. The half-life of ALP duplicated genes was estimated to be 0.87 Myr. Strong purifying selection acting on the ancestral gene ALP-A1 was undiminished by the evolution of duplicated genes. The evolution of the ALP family shows that repeated elements facilitate both gene duplication and expression of duplicated genes and highlights their importance for the evolution of gene repertoire in large plant genomes. PMID- 17135335 TI - Levels of bimatoprost acid in the aqueous humour after bimatoprost treatment of patients with cataract. AB - AIM: To determine the aqueous humour concentration of the acid hydrolysis products of bimatoprost and latanoprost after a single topical dose of bimatoprost 0.03% or latanoprost 0.005% in humans. METHODS: Randomised, controlled, double-masked, prospective study. 48 eyes of 48 patients scheduled for routine cataract surgery were randomised in an 8:2:2 ratio to treatment with a single 30 mul drop of bimatoprost 0.03%, latanoprost 0.005% or placebo at 1, 3, 6 or 12 h before the scheduled cataract surgery. Aqueous humour samples were withdrawn at the beginning of the surgical procedure and analysed using high performance liquid chromatography-tandem mass spectrometry. RESULTS: Bimatoprost acid (17-phenyl trinor prostaglandin F2alpha) was detected in aqueous samples at a mean concentration of 5.0 nM at hour 1, 6.7 nM at hour 3 and 1.9 nM at hour 6 after bimatoprost treatment. After latanoprost treatment, the mean concentration of latanoprost acid (13,14-dihydro-17-phenyl trinor prostaglandin F2alpha) in aqueous samples was 29.1 nM at hour 1, 41.3 nM at hour 3 and 2.5 nM at hour 6. Acid metabolites were below the limit of quantitation in all samples taken 12 h after dosing and in all samples from placebo-treated patients. None of the samples from latanoprost-treated patients contained quantifiable levels of non metabolised latanoprost. Non-metabolised bimatoprost was detected in aqueous samples at a mean concentration of 6.6 nM at hour 1 and 2.4 nM at hour 3 after bimatoprost treatment. CONCLUSIONS: Low levels of bimatoprost acid were detected in aqueous humour samples from patients with cataract treated with a single dose of bimatoprost. Latanoprost acid concentrations in samples from patients treated with latanoprost were at least sixfold higher. These results suggest that bimatoprost acid in the aqueous humour does not sufficiently account for the ocular hypotensive efficacy of bimatoprost. PMID- 17135336 TI - Vitreoretinal morphology in active ocular toxoplasmosis: a prospective study by optical coherence tomography. AB - AIM: To investigate the third generation optical coherence tomography (OCT3) findings in patients with active ocular toxoplasmosis. METHODS: A prospective observational case series, including 15 patients with active ocular toxoplasmosis in at least one eye evaluated at a single centre. Vitreoretinal morphological features at baseline and changes within a 24-week follow-up interval on OCT3 were evaluated. RESULTS: The active ocular toxoplasmosis lesion was classified clinically as punctate (n = 6), focal (n = 6) or satellite (n = 3). Retinal layers were hyper-reflective at the active lesion site, and some degree of retinal pigment epithelium-choriocapillaris/choroidal optical shadowing was seen in all patients. In general, the retina was thinned at the active lesion site in eyes with punctate lesions and thickened in eyes with focal and satellite lesions. When detected by OCT3, the posterior hyaloid appeared thickened. While focally detached over punctate lesions, the posterior hyaloid was partially detached, but still attached to the lesion in focal and satellite lesions. Additional findings (not detected on clinical examination) include diffuse macular oedema (n = 6), vitreomacular traction (n = 3) and maculoschisis (n = 1). During follow-up, a decrease in retinal thickness and focal choriocapillaris/choroidal relative hyper-reflectivity were observed at the former lesion site, and posterior vitreous detachment progressed/occurred in all patients. CONCLUSION: OCT3 enabled identification of morphological features underestimated on clinical examination in patients with ocular toxoplasmosis, which may expand the clinical spectrum of the disease. Further studies are needed to verify the relevance of OCT3 in assisting with the diagnosis and management of ocular toxoplasmosis. PMID- 17135337 TI - Bupivacaine injection of eye muscles to treat strabismus. AB - BACKGROUND: Bupivacaine injected into animal muscles induces a cycle of myotoxicity, degeneration, regeneration and hypertrophy of muscle fibres, without adverse effects on other tissues. This induced hypertrophy can be harnessed to treat strabismus. METHODS: Bupivacaine, 4.5 ml of a 0.75% solution, was injected into the right lateral rectus (RLR) muscle of a patient who had diplopia and who showed 14-prism-dioptres oesotropia. RESULTS: RLR paresis persisted for 7 days. Then, the RLR regained its abducting ability, and progressive improvement of alignment to 4-prism-dioptres oesophoria occurred over the next 33 days, with the elimination of diplopia. Alignment remained the same at 54 days after injection. Magnetic resonance imaging showed a focal increase in the size of the injected RLR of 58% in the posterior area, with reduced change in anterior portions of the RLR. CONCLUSION: Injection of bupivacaine to induce hypertrophy of the injected muscle and thus alter eye alignment was effective in our patient. This approach can be a useful addition to the treatment of strabismus. PMID- 17135338 TI - Management of orbital xanthogranuloma with methotrexate. AB - BACKGROUND/AIMS: To evaluate the efficacy of methotrexate (MTX) treatment for the periorbital findings in adult-onset xanthogranuloma (AOX). METHODS: The medical records of three patients with AOX, with and without asthma, who were treated with MTX at Oregon Health & Science University, Portland, Oregon, USA were examined. Diagnosis of AOX was made by biopsy in all patients. The patients were evaluated between February 1998 and July 2006. All patients had failed prior medical and/or surgical treatment. MTX was administered at 10-20 mg/week with folate supplementation and a course of corticosteroids. Efficacy was assessed on the basis of improvement in skin discoloration, involvement of the visual axis and patients' report of inflammation. RESULTS: All three patients were started on MTX, but one patient discontinued treatment after 3 weeks due to nausea. With follow-up as long as 3 years, the two patients who continued treatment lost the yellow discoloration of their skin, and they reported significantly less inflammation and ptosis after treatment. Oral corticosteroids could be reduced or discontinued. CONCLUSIONS: AOX is a rare, persistent disease that commonly involves the preseptal fat. MTX is a therapeutic option for this illness. PMID- 17135339 TI - Correlations in refractive errors between siblings in the Singapore Cohort Study of Risk factors for Myopia. AB - BACKGROUND: The prevalence of myopia in parts of South East Asia has risen dramatically over the past 1-2 generations, suggesting that environmental factors may be particularly important determinants of refractive development in these populations. AIM: To assess the contribution of familial factors (shared genes and/or shared family environment) to refractive error and ocular component dimensions of school-aged children in Singapore. METHODS: Data were available for 315 children who had one or more siblings also participating in the Singapore Cohort Study of the Risk factors for Myopia (SCORM). Refractive error and ocular biometric parameters were measured under cycloplegia at baseline when children were 7-9 years, and at yearly follow-up sessions for the next 3 years, using consistent clinical procedures. The time children spent performing a variety of nearwork-related tasks was obtained from questionnaires. Familial influences were assessed by calculating between-sibling correlations. RESULTS: After adjusting for age and sex, the between-sibling correlation in refractive error was 0.447 (95% CI 0.314 to 0.564), suggesting that familial factors account for 63-100% of the variation in the cohort. The between-sibling correlation for 1-year change in refractive error was similarly high, at 0.420 (95% CI 0.282 to 0.543). All ocular component dimensions were correlated significantly between siblings, especially for corneal curvature and vitreous chamber depth--the major structural determinants of refraction. The amount of time siblings spent engaged in nearwork tasks (reading, watching TV, playing video games, computing) and in outdoor activities was also highly correlated between siblings (p<0.001). CONCLUSION: Shared genes and/or shared environment are important factors in the refractive development of children in Singapore. Because the time spent in nearwork tasks is highly correlated between siblings, epidemiological studies will benefit from precise, quantitative measures of refractive error in parents and more distant relatives in order to begin to dissociate genetic and environmental sources of variation. PMID- 17135340 TI - Evaluation of a new soft tipped injector for the implantation of foldable intraocular lenses. AB - AIM: To evaluate the R-INJ-04 soft-tipped injector, a new injector with an integral round nozzle manufactured by Rayner Intraocular Lenses, England. METHODS: 16 Rayner C-flex intraocular lenses (IOLs; Rayner Intraocular lenses, England) ranging between +10 and +30 D (2 for each power) were tested. An ophthalmic viscoelastic device (Healon, AMO, Santa Ana, California, USA) was applied to the injectors. The IOLs were loaded according to the company injector's instructions for use and were injected into a Petri dish. After the injection, all the IOLs and nozzles were evaluated by gross (macroscopic) and microscopic analyses and photographed under a light microscope. One lens of each power and the cartridge used for the implantation were then sent for further analysis by scanning electron microscopy (SEM). The rest of the IOLs were tested for power and modulation transfer function (MTF). RESULTS: All the injections were successful. No damage to the IOLs or to the injectors was found by gross examination, light microscopy and SEM. No deposits were found on the IOL optical surfaces or haptics. Power and MTF analysis showed a close match with the original measurements. CONCLUSION: Our results suggest that the R-INJ-04 soft tipped injector is safe for the implantation of the C-flex IOL with power range from 10 to 30 D. No structural damage to the IOLs or to the injectors was found, and the lens power and light transmission properties were not damaged in any way by the injection process. PMID- 17135341 TI - S-Nitrosylation of human variant albumin Liprizzi (R410C) confers potent antibacterial and cytoprotective properties. AB - The S-nitrosylated forms of certain proteins such as albumin have been thought to be circulating endogenous reservoirs of nitric oxide (NO) and may have potential as NO donors in therapeutic applications. In this study, we investigated the characteristics of R410C, a genetic variant of human serum albumin with two free thiols at positions 34 (Cys-34) and 410 (Cys-410), as a NO carrier via S-nitroso formation. A biotin switch assay revealed that Cys-410 was more rapidly and efficiently nitrosylated than was Cys-34. Nitrosylation of Cys-410 introduced only small conformational changes in the protein, which were detected by far-UV circular dichroism but not by near-UV circular dichroism. In addition, both native R410C and S-nitrosylated R410C did not induce molecular heterogeneity through oligomerization. S-Nitrosylated R410C exhibited strong antibacterial activity against Salmonella typhimurium in vitro and suppressed apoptosis of U937 human promonocytic cells induced by Fas ligand. In a rat ischemia-reperfusion liver injury model, S-nitrosylated R410C treatment significantly reduced liver damage, as indicated by markedly decreased release of liver enzymes (aspartate aminotransferase and alanine aminotransferase). Pharmacokinetic analyses indicated retention of the S-nitroso moiety of S-nitrosylated R410C in circulation after i.v. injection, with an approximate half-life of 20.4 min in the mouse. These data suggest that R410C can be a useful NO carrier and can be regarded as a new class of S-nitrosylated proteins possessing antibacterial and cytoprotective properties with a circulation time sufficient for in vivo biological activity. PMID- 17135342 TI - Selective clearance of macrophages in atherosclerotic plaques by the protein synthesis inhibitor cycloheximide. AB - Macrophages are an essential component of unstable atherosclerotic plaques and play a pivotal role in the destabilization process. We have demonstrated previously that local delivery of the mammalian target of rapamycin (mTOR) inhibitor everolimus selectively clears macrophages in rabbit plaques. Because mTOR controls mRNA translation, inhibition of protein synthesis might induce selective macrophage cell death. We therefore investigated in the present study the effect of the protein synthesis inhibitor cycloheximide on macrophage and smooth muscle cell (SMC) viability. In vitro studies with cultured macrophages and SMCs showed that cycloheximide induced selective apoptosis of macrophages in a concentration- and time-dependent manner. Moreover, macrophages could be selectively depleted in rabbit carotid artery rings with collar-induced atherosclerotic plaques after in vitro treatment with cycloheximide. Local in vivo administration of cycloheximide via osmotic minipumps to rabbit carotid arteries with collar-induced atherosclerotic plaques significantly reduced the macrophage but not the SMC content. Cycloheximide-treated plaques showed signs of apoptosis (increased terminal deoxynucleotidyl transferase end labeling and fluorescein isothiocyanate-Val-Ala-dl-Asp(O-methyl)-fluoromethylketone labeling) that did not colocalize with SMCs. Organ chamber studies demonstrated that the functionality of SMCs and the endothelium were not influenced by cycloheximide treatment. All together, these findings demonstrate that cycloheximide decreases the macrophage load in atherosclerotic plaques by induction of apoptosis without changing SMC content or contractility. PMID- 17135343 TI - The hypolipidemic agent guggulsterone regulates the expression of human bile salt export pump: dominance of transactivation over farsenoid X receptor-mediated antagonism. AB - Conversion of cholesterol to bile acids in the liver is initiated by the rate limiting enzyme cholesterol 7alpha-hydroxylase (CYP7A1) and excretion of bile acids from the liver is mediated by the bile salt export pump (BSEP). The expression of CYP7A1 and BSEP is coordinately regulated by a negative feedback and positive feed-forward mechanism, respectively, through bile acid-mediated activation of farsenoid X receptor (FXR). It is well established that hypolipidemic agent guggulsterone is an FXR antagonist and down-regulates FXR target genes. In this study, however, we have demonstrated that guggulsterone synergistically induced the expression of BSEP in cells treated with FXR agonist bile acids. A dissection study located in the BSEP promoter an activating protein (AP)-1 site supporting the action of guggulsterone. Deletion or mutation of the AP-1 element was diminished, whereas insertion of the AP-1 element into a heterologous promoter enhanced activation of the promoter by guggulsterone. Selective c-Jun N-terminal kinase and extracellular signal-regulated kinase inhibitors markedly decreased the transactivation, suggesting an involvement of AP-1 activation pathway in the up-regulation of BSEP by guggulsterone. Consistent with its FXR antagonism, guggulsterone antagonized bile acid-mediated transactivation of BSEP promoter when the AP-1 element was disrupted. In conclusion, guggulsterone regulates BSEP expression through composite mechanisms, and the transactivation through the AP-1 element is dominant over the FXR mediated antagonism. The up-regulation of BSEP expression by guggulsterone without activating FXR pathway as an FXR agonist to suppress CYP7A1 expression represents a possible mechanism for guggulsterone-mediated hypolipidemic effect. PMID- 17135344 TI - Down-regulation of intestinal drug transporters in chronic renal failure in rats. AB - Chronic renal failure (CRF) is associated with an increased bioavailability of drugs by a poorly understood mechanism. One hypothesis is a reduction in the elimination of drugs by the intestine, i.e., drug elimination mediated by protein membrane transporters such as P-glycoprotein (Pgp) and multidrug-resistance related protein (MRP) 2. The present study aimed to investigate the repercussions of CRF on intestinal transporters involved in drug absorption [organic anion transportingpolypeptide (Oatp)] and those implicated in drug extrusion (Pgp and MRP2). Pgp, MRP2, MRP3, Oatp2, and Oatp3 protein expression and Pgp, MRP2, and Oatp3 mRNA expression were assessed in the intestine of CRF (induced by five sixth nephrectomy) and control rats. Pgp and MRP2 activities were measured using the everted gut technique. Rat enterocytes and Caco-2 cells were incubated with sera from control and CRF rats to characterize the mechanism of transporters' down-regulation. Protein expression of Pgp, MRP2, and MRP3 were reduced by more than 40% (p < 0.01) in CRF rats, whereas Oatp2 and Oatp3 expression remained unchanged. There was no difference in the mRNA levels assessed by real-time polymerase chain reaction. Pgp and MRP2 activities were decreased by 30 and 25%, respectively, in CRF rats compared with control (p < 0.05). Uremic sera induced a reduction in protein expression and in activity of drug transporters compared with control sera. Our results demonstrate that CRF in rats is associated with a decrease in intestinal Pgp and MRP2 protein expression and function secondarily to serum uremic factors. This reduction could explain the increased bioavailability of drugs in CRF. PMID- 17135345 TI - Defective taxane stimulation of epirubicinol formation in the human heart: insight into the cardiac tolerability of epirubicin-taxane chemotherapies. AB - The antitumor anthracycline doxorubicin induces a dose-related cardiotoxicity that correlates with the myocardial levels of its secondary alcohol metabolite doxorubicinol. Combining doxorubicin with taxanes such as paclitaxel or docetaxel may aggravate cardiotoxicity, presumably because the taxanes cause an allosteric like stimulation of cytoplasmic aldehyde reductases that convert doxorubicin to doxorubicinol in the heart. A less severe aggravation of cardiotoxicity was observed on combining taxanes with epirubicin, a closely related analog of doxorubicin; therefore, we characterized whether the cardiac tolerability of epirubicin-taxane therapies could be due to a defective taxane stimulation of the conversion of epirubicin to its secondary alcohol metabolite epirubicinol. Comparisons between doxorubicin and epirubicin in isolated human heart cytosol showed that epirubicin exhibited a lower V(max)/K(m) value for reaction with aldehyde reductases and a defective stimulation of epirubicinol formation by paclitaxel or docetaxel. A similar pattern occurred in the soluble fraction of human myocardial strips incubated in plasma with anthracyclines and paclitaxel or docetaxel, formulated in their clinical vehicles Cremophor EL or polysorbate 80. Doxorubicin, but not epirubicin, was also able to generate reactive oxygen species in the membrane fraction of myocardial strips; however, the levels of doxorubicin-derived reactive oxygen species were not further augmented by paclitaxel. These results support the notion that taxanes might aggravate the cardiotoxicity of doxorubicin through a specific stimulation of doxorubicinol formation. The failure of paclitaxel or docetaxel to stimulate epirubicinol formation therefore uncovers an important determinant of the improved cardiac tolerability of epirubicin-taxane combinations. PMID- 17135346 TI - Niemann-Pick C1-like 1 overexpression facilitates ezetimibe-sensitive cholesterol and beta-sitosterol uptake in CaCo-2 cells. AB - Previous in vivo studies including those with knockout mice suggested that Niemann-Pick C1-like 1 (NPC1L1) plays an essential role in the intestinal absorption of cholesterol. To characterize the mechanism of cholesterol uptake mediated by NPC1L1, an in vitro system reflecting the function of this transporter needs to be established. In the present study, we constructed NPC1L1 overexpressing CaCo-2 cells as an in vitro model and characterized the transport properties of NPC1L1. Immunohistochemical staining revealed that CaCo-2 cells express NPC1L1 on the apical membrane. It was also demonstrated that the uptakes of both cholesterol and beta-sitosterol are increased by NPC1L1 overexpression. In addition, the uptake of cholesterol was increased in a dose-dependent manner by an increase in the content of taurocholate in micelles, whereas micellar phosphatidylcholine showed a negative correlation with cholesterol uptake. Furthermore, it was confirmed that sterol uptake increased by NPC1L1 overexpression was inhibited by ezetimibe. We could thus establish an in vitro intestinal model to study the mechanism of NPC1L1-dependent sterol uptake and to screen drug candidates whose target is NPC1L1. PMID- 17135347 TI - Habituation deficits induced by metabotropic glutamate receptors 2/3 receptor blockade in mice: reversal by antipsychotic drugs. AB - Cortical metabotropic glutamate receptors (mGluRs) seem to be involved in habituation of simple stimulus-bound behaviors (e.g., habituation to acoustic startle or odor-elicited orienting response). Habituation deficits may contribute to the cognitive symptoms of schizophrenia. In the present study, male NMRI mice were injected with mGluR2/3 antagonist 2S-2-amino-2-(1S,2S-2-carboxycyclopropyl-1 yl)-3-(xanth-9-yl)propanoic acid (LY-341495) 30 min before being placed into novel arenas for automatic motor activity recording (2-h sessions). Administration of LY-341495 (1-10 mg/kg s.c.) dose-dependently prevented the habituation of the locomotor activity. Effects of LY-341495 (10 mg/kg) were fully and dose-dependently reversed by i.p. administration of haloperidol (0.03-0.3 mg/kg), clozapine (1-10 mg/kg), risperidone (0.01-0.1 mg/kg), olanzapine (0.3-3 mg/kg), aripiprazole (1-10 mg/kg), and sulpiride (3-30 mg/kg), each of which was given 15 min before the test. Effects of antipsychotic drugs were observed at the dose levels that did not affect spontaneous motor activity. LY-341495-induced delayed hyperactivity was also partially attenuated by lithium (50-200 mg/kg), amisulpride (1-10 mg/kg), and the selective dopamine D3 antagonist trans-N-[4-[2 (6-cyano-1,2,3,4-tetrahydroisoquinolin-2-yl)ethyl]cyclohexyl]-4 quinolinecarboxamide (SB-277011A; 3-30 mg/kg). Application of diazepam, imipramine, or several agonists and/or antagonists acting at various receptors that are thought to be relevant for antipsychotic treatment [e.g., 5 hydroxytryptamine (5-HT)(2A), 5-HT(3), and 5-HT(6) antagonists; 5-HT(1A) agonist; D4 antagonist; CB1 antagonist; ampakines; and glycine transporter inhibitor) had no appreciable effects. Thus, behavioral deficits induced by mGluR2/3 blockade (such as delayed motor hyperactivity) are selectively reversed by clinically used antipsychotic drugs. PMID- 17135348 TI - MicroRNA-206 colocalizes with ribosome-rich regions in both the nucleolus and cytoplasm of rat myogenic cells. AB - MicroRNAs are small, approximately 21- to 24-nt RNAs that have been found to regulate gene expression. miR-206 is a microRNA that is expressed at high levels in Drosophila, zebrafish, and mouse skeletal muscle and is thought to be involved in the attainment and/or maintenance of the differentiated state. We used locked nucleic acid probes for in situ hybridization analysis of the intracellular localization of miR-206 during differentiation of rat myogenic cells. Like most microRNAs, which are presumed to suppress translation of target mRNAs, we found that miR-206 occupies a cytoplasmic location in cultured myoblasts and differentiated myotubes and that its level increases in myotubes over the course of differentiation, consistent with previous findings in muscle tissue in vivo. However, to our surprise, we also observed miR-206 to be concentrated in nucleoli. A probe designed to be complementary to the precursor forms of miR-206 gave no nucleolar signal. We characterized the intracellular localization of miR 206 at higher spatial resolution and found that a substantial fraction colocalizes with 28S rRNA in both the cytoplasm and the nucleolus. miR-206 is not concentrated in either the fibrillar centers of the nucleolus or the dense fibrillar component, where ribosomal RNA transcription and early processing occur, but rather is localized in the granular component, the region of the nucleolus where final ribosome assembly takes place. These results suggest that miR-206 may associate both with nascent ribosomes in the nucleolus and with exported, functional ribosomes in the cytoplasm. PMID- 17135349 TI - Genomewide screens for Escherichia coli genes affecting growth of T7 bacteriophage. AB - Use of bacteriophages as a therapy for bacterial infection has been attempted over the last century. Such an endeavor requires the elucidation of basic aspects of the host-virus interactions and the resistance mechanisms of the host. Two recently developed bacterial collections now enable a genomewide search of the genetic interactions between Escherichia coli and bacteriophages. We have screened >85% of the E. coli genes for their ability to inhibit growth of T7 phage and >90% of the host genes for their ability to be used by the virus. In addition to identifying all of the known interactions, several other interactions have been identified. E. coli CMP kinase is essential for T7 growth, whereas overexpression of the E. coli uridine/cytidine kinase inhibits T7 growth. Mutations in any one of nine genes that encode enzymes for the synthesis of the E. coli lipopolysaccharide receptor for T7 adsorption leads to T7 resistance. Selection of T7 phage that can recognize these altered receptors has enabled the construction of phage to which the host is 100-fold less resistant. PMID- 17135350 TI - Human cytomegalovirus pUL37x1 induces the release of endoplasmic reticulum calcium stores. AB - The human CMV UL37x1-encoded protein, also known as the viral mitochondria localized inhibitor of apoptosis, traffics to the endoplasmic reticulum and mitochondria of infected cells. It induces the fragmentation of mitochondria and blocks apoptosis. We demonstrate that UL37x1 protein mobilizes Ca(2+) from the endoplasmic reticulum into the cytosol. This release is accompanied by cell rounding, cell swelling, and reorganization of the actin cytoskeleton, and these morphological changes can be substantially blocked by a Ca(2+) chelating agent. The UL37x1-mediated release of Ca(2+) from the endoplasmic reticulum likely has multiple consequences, including induction of the unfolded protein response, modulation of mitochondrial function, induction of mitochondrial fission, and protection against apoptotic stimuli. PMID- 17135351 TI - Mammalian pharmacokinetics of carbon nanotubes using intrinsic near-infrared fluorescence. AB - Individualized, chemically pristine single-walled carbon nanotubes have been intravenously administered to rabbits and monitored through their characteristic near-infrared fluorescence. Spectra indicated that blood proteins displaced the nanotube coating of synthetic surfactant molecules within seconds. The nanotube concentration in the blood serum decreased exponentially with a half-life of 1.0 +/- 0.1 h. No adverse effects from low-level nanotube exposure could be detected from behavior or pathological examination. At 24 h after i.v. administration, significant concentrations of nanotubes were found only in the liver. These results demonstrate that debundled single-walled carbon nanotubes are high contrast near-infrared fluorophores that can be sensitively and selectively tracked in mammalian tissues using optical methods. In addition, the absence of acute toxicity and promising circulation persistence suggest the potential of carbon nanotubes in future pharmaceutical applications. PMID- 17135352 TI - Note on the energy density in the solvent induced by a solute. AB - The question of how far the effect of the presence of a solute molecule propagates into the solvent is studied in a lattice model that had been used earlier to describe hydrophobic interactions. The local energy density in the model solvent is obtained as an explicit function of distance from the solute and is found to decay to its bulk-phase value with the same decay length as that of the solvent-mediated part of the potential of mean force between a pair of solute molecules. The integrated deviation of the energy density from its bulk-phase value is evaluated in Bethe-Guggenheim approximation and shown to be identical to the energy change accompanying the dissolution of the solute as obtained from the temperature dependence of its solubility. PMID- 17135353 TI - Dynamics of allosteric transitions in GroEL. AB - The chaperonin GroEL-GroES, a machine that helps proteins to fold, cycles through a number of allosteric states, the T state, with high affinity for substrate proteins, the ATP-bound R state, and the R" (GroEL-ADP-GroES) complex. Here, we use a self-organized polymer model for the GroEL allosteric states and a general structure-based technique to simulate the dynamics of allosteric transitions in two subunits of GroEL and the heptamer. The T --> R transition, in which the apical domains undergo counterclockwise motion, is mediated by a multiple salt bridge switch mechanism, in which a series of salt-bridges break and form. The initial event in the R -->R" transition, during which GroEL rotates clockwise, involves a spectacular outside-in movement of helices K and L that results in K80 D359 salt-bridge formation. In both the transitions there is considerable heterogeneity in the transition pathways. The transition state ensembles (TSEs) connecting the T, R, and R" states are broad with the TSE for the T --> R transition being more plastic than the R --> R" TSE. PMID- 17135354 TI - Sleep deprivation inhibits adult neurogenesis in the hippocampus by elevating glucocorticoids. AB - Prolonged sleep deprivation is stressful and has been associated with adverse consequences for health and cognitive performance. Here, we show that sleep deprivation inhibits adult neurogenesis at a time when circulating levels of corticosterone are elevated. Moreover, clamping levels of this hormone prevents the sleep deprivation-induced reduction of cell proliferation. The recovery of normal levels of adult neurogenesis after chronic sleep deprivation occurs over a 2-wk period and involves a temporary increase in new neuron formation. This compensatory increase is dissociated from glucocorticoid levels as well as from the restoration of normal sleep patterns. Collectively, these findings suggest that, although sleep deprivation inhibits adult neurogenesis by acting as a stressor, its compensatory aftereffects involve glucocorticoid-independent factors. PMID- 17135355 TI - Developmental plasticity mirrors differences among taxa in spadefoot toads linking plasticity and diversity. AB - Developmental plasticity is found in most organisms, but its role in evolution remains controversial. Environmentally induced phenotypic differences may be translated into adaptive divergence among lineages experiencing different environmental conditions through genetic accommodation. To examine this evolutionary mechanism, we studied the relationship between plasticity in larval development, postmetamorphic morphology, and morphological diversity in spadefoot toads, a group of closely related species that are highly divergent in the larval period and body shape and are distributed throughout temperate areas of both the New and the Old World. Previous studies showed that accelerated metamorphosis is adaptive for desert-dwelling spadefoot toads. We show that even under common garden conditions, spadefoot toad species show divergent reaction norms for the larval period. In addition, experimentally induced changes in the larval period caused correlated morphological changes in postmetamorphic individuals such that long larval periods resulted in relatively longer hindlimbs and snouts. A comparative analysis of morphological variation across spadefoot toad species also revealed a positive correlation between the larval period and limb and snout lengths, mirroring the effects of within-species plasticity at a higher taxonomic level. Indeed, after approximately 110 Ma of independent evolution, differences in the larval period explain 57% of the variance in relative limb length and 33% of snout length across species. Thus, morphological diversity across these species appears to have evolved as a correlated response to selection for a reduced larval period in desert-dwelling species, possibly diverging from ancestral plasticity through genetic accommodation. PMID- 17135356 TI - A synthetic phage lambda regulatory circuit. AB - Analysis of synthetic gene regulatory circuits can provide insight into circuit behavior and evolution. An alternative approach is to modify a naturally occurring circuit, by using genetic methods to select functional circuits and evolve their properties. We have applied this approach to the circuitry of phage lambda. This phage grows lytically, forms stable lysogens, and can switch from this regulatory state to lytic growth. Genetic selections are available for each behavior. We previously replaced lambda Cro in the intact phage with a module including Lac repressor, whose function is tunable with small molecules, and several cis-acting sites. Here, we have in addition replaced lambda CI repressor with another tunable module, Tet repressor and several cis-acting sites. Tet repressor lacks several important properties of CI, including positive autoregulation and cooperative DNA binding. Using a combinatorial approach, we isolated phage variants with behavior similar to that of WT lambda. These variants grew lytically and formed stable lysogens. Lysogens underwent prophage induction upon addition of a ligand that weakens binding by the Tet repressor. Strikingly, however, addition of a ligand that weakens binding by Lac repressor also induced lysogens. This finding indicates that Lac repressor was present in the lysogens and was necessary for stable lysogeny. Therefore, these isolates had an altered wiring diagram from that of lambda. We speculate that this complexity is needed to compensate for the missing features. Our method is generally useful for making customized gene regulatory circuits whose activity is regulated by small molecules or protein cofactors. PMID- 17135357 TI - Protein kinase CK2 acts as a signal molecule switching between the NDPK-A/AMPK alpha1 complex and NDPK-B. AB - Previously we elucidated the molecular interaction between the nucleoside diphosphate kinase A (NDPK-A)/AMP-activated protein kinase (AMPK) alpha1 complex, discovering a process we termed "substrate channeling." Here, we investigate the protein-protein interaction of the substrate channeling complex with the pleiotropic protein kinase, CK2 (formerly casein kinase 2). We show that CK2 is part of the NDPK-A/AMPK alpha1 complex under basal (background AMPK activity) conditions, binding directly to each of the complex components independently. We report that when S122 on NDPK-A is phosphorylated by AMPK alpha1 in vivo, (i.e., stimulation of AMPK using either metformin or phenformin) initiating the substrate channeling mechanism, the catalytic subunit of CK2 (CK2alpha) is expelled from the complex and translocates to bind NDPK-B, a closely related but independent isoform of NDPK. Thus, we find that the AMPK-dependent phospho-status of S122 on NDPK-A determines whether CK2alpha swaps partners between NDPK-A and NDPK-B. This is the first reported linkage between NDPK-A and NDPK-B via a phosphorylation pathway and could explain the complex biology of NDPK. This study also offers an explanation as to how CK2alpha exclusion mutations (S120A or S122D of NDPK-A) on NDPK-A might have implications in cancer biology and general cellular energy metabolism. PMID- 17135358 TI - Zinc chelation inhibits HIV Vif activity and liberates antiviral function of the cytidine deaminase APOBEC3G. AB - APOBEC3 proteins are cellular antiviral proteins that are targeted for proteasomal degradation by primate lentiviral Vif proteins. Vif acts as a substrate receptor for the Cullin5 (Cul5) E3 ubiquitin ligase, specifically interacting with Cul5 through a novel H-(x5)-C-(x17-18)-C-(x3-5)-H zinc binding motif. Using the membrane-permeable zinc chelator, N,N,N',N'-tetrakis-(2 pyridylmethyl) ethylenediamine (TPEN), we demonstrated a requirement for zinc for Vif function in vivo. Treatment with TPEN at an IC50 of 1.79 microM inhibits Cul5 recruitment and APOBEC3G (A3G) degradation. Zinc chelation prevented Vif function in infectivity assays, allowing the virus to become sensitive to the antiviral activity of A3G. Zinc chelation had no effect on cellular Cul5-SOCS3 E3 ligase assembly, suggesting that zinc-dependent E3 ligase assembly may be unique to HIV 1 Vif, representing a new target for novel drug design. PMID- 17135359 TI - Genetic and pharmacological inhibition of GITR-GITRL interaction reduces chronic lung injury induced by bleomycin instillation. AB - We have recently identified a gene named GITR (glucocorticoid-induced TNF receptor related gene). GITR is expressed in different cells and tissues such as T lymphocytes from thymus and spleen and lymph nodes, and also in the lung. GITR ligand (GITRL) is expressed in several cells including macrophages, B cells, dendritic cells, and endothelial cells. In the present study, by comparing the responses in wild-type (WT) mice (GITR+/+) and GITR-deficient mice (GITR-/-), we investigated the role played by GITR-GITRL interaction in the development of chronic lung injury caused by bleomycin instillation. When compared with bleomycin-treated GITR+/+ mice, bleomycin-treated GITR-/- mice exhibited a reduced degree of i) lung infiltration with polymorphonuclear neutrophils (MPO activity); ii) edema formation; iii) histological evidence of lung injury; iv) TNF-alpha and interleukin (IL)-1beta production; v) nitrotyrosine formation; and vi) NF-kappaB activation. The cotreatment of GITR+/+ mice with Fc-GITR fusion protein (6.25 microg/mouse) also significantly attenuated all of the above indicators of lung damage and inflammation. Our results clearly demonstrate that GITR-GITRL interaction plays an important role in the chronic lung injury induced by bleomycin in the mice. PMID- 17135360 TI - Mammalian dap3 is an essential gene required for mitochondrial homeostasis in vivo and contributing to the extrinsic pathway for apoptosis. AB - Death-associated protein-3 (DAP3) is a GTP binding protein previously implicated in both intramitochondrial protein synthesis and apoptosis. To explore the in vivo roles of DAP3, we generated and characterized DAP3-deficient mice. Homozygous dap3-/- embryos died at approximately day 9.5 in utero. The dap3-/- embryos and placentas were markedly shrunken. Embryos had arrested development, displaying severe growth restriction and lack of axial turning. Transmission electron microscopy analysis revealed abnormal, shrunken mitochondria with swollen crystae in dap3-/- embryos. Levels of cytochrome c oxidase-I, a protein encoded in the mitochondrial genome, were reduced in dap3-/- embryos, consistent with a role for DAP3 in intramitochondrial protein synthesis. A requirement for DAP3 in mitochondrial respiration was also revealed by oxygen consumption measurements using cultured cells treated with DAP3-specific small interfering RNA (siRNA). Studies of cultured cells from dap3-/- embryos confirmed a role in apoptosis induced by stimuli that trigger the extrinsic (TNFalpha, TRAIL, anti Fas antibody) but not intrinsic (mitochondrial) cell death pathway. Thus, DAP3 joins a growing list of bifunctional proteins that play roles in normal mitochondrial physiology and in apoptosis. PMID- 17135361 TI - Dissecting the signaling pathway of nicotine-mediated neuroprotection in a mouse Alzheimer disease model. AB - Nicotine has a therapeutic benefit in treating Alzheimer's disease (AD). In the present study we show that nicotine decreases accumulation of beta-amyloid (Abeta) in the cortex and hippocampus of APP (V717I) transgenic mice. Nicotine prevents activation of NF-kappaB and c-Myc by inhibiting the activation of MAP kinases (MAPKs). As a result, the activity of inducible NOS and the production of NO are down-regulated. RNA interference experiments show that the above nicotine mediated process requires alpha7 nAChR. Nicotine decreases Abeta via the activation of alpha7nAChRs through MAPK, NF-kappaB, and c-myc pathways. Nicotine also inhibits apoptosis and cell cycle progression in this mouse line. The dissected signaling pathway of nicotine-mediated neuroprotection in the present study provides a mechanistic basis for the potential development of drug targets for treating AD. PMID- 17135362 TI - Insidious adrenocortical insufficiency underlies neuroendocrine dysregulation in TIF-2 deficient mice. AB - The transcription-intermediary-factor-2 (TIF-2) is a coactivator of the glucocorticoid receptor (GR), and its disruption would be expected to influence glucocorticoid-mediated control of the hypothalamo-pituitary-adrenal (HPA) axis. Here, we show that its targeted deletion in mice is associated with altered expression of several glucocorticoid-dependent components of HPA regulation (e.g., corticotropin-releasing hormone, vasopressin, ACTH, glucocorticoid receptors), suggestive of hyperactivity under basal conditions. At the same time, TIF-2(-/-) mice display significantly lower basal corticosterone levels and a sluggish and blunted initial secretory response to brief emotional and prolonged physical stress. Subsequent analysis revealed this discrepancy to result from pronounced aberrations in the structure and function of the adrenal gland, including the cytoarchitectural organization of the zona fasciculata and basal and stress-induced expression of key elements of steroid hormone synthesis, such as the steroidogenic acute regulatory (StAR) protein and 3beta-hydroxysteroid dehydrogenase (3beta-HSD). In addition, altered expression levels of two nuclear receptors, DAX-1 and steroidogenic factor 1 (SF-1), in the adrenal cortex strengthen the view that TIF-2 deletion disrupts adrenocortical development and steroid biosynthesis. Thus, hyperactivity of the hypothalamo-pituitary unit is ascribed to insidious adrenal insufficiency and impaired glucocorticoid feedback. PMID- 17135363 TI - Roles and origins of leukocyte lipid bodies: proteomic and ultrastructural studies. AB - Lipid bodies (LBs), multifunctional organelles present in most eukaryotic cells, are sites of eicosanoid formation in leukocytes; but little is known about the composition of leukocyte LBs or the biogenesis and internal structures of LBs from mammalian cells. Proteomic analyses of LBs purified from human monocytic U937 cells detected, common to LBs in other cells, proteins involved in cholesterol and triglyceride metabolism, Rab GTPases, and many membrane and endoplasmic reticulum (ER)-associated proteins. Newly lipid body (LB)-associated proteins included MRP-14, potentially involved in arachidonate transport, and ribosomal subunit proteins and translation regulatory proteins. Ultrastructurally, in U937 cells as well as human neutrophils and eosinophils, ribosomes are attached to and distributed within LBs, and LBs contain extensive ER-like membranes. The presence of ribosomes, ER-like membranes and many membrane associated and ER luminal proteins within LBs, supports a new model by which enveloped ER-membranes and domains form LBs and indicates that LBs may be sites of protein synthesis. PMID- 17135364 TI - Amelioration of diabetes by imatinib mesylate (Gleevec): role of beta-cell NF kappaB activation and anti-apoptotic preconditioning. AB - It was recently reported that tyrosine kinase inhibitor imatinib mesylate (Gleevec) improves Type 2 diabetes, possibly by decreasing insulin resistance. However, as both Type 2 and Type 1 diabetes are characterized by beta-cell dysfunction and death, we investigated whether imatinib counteracts diabetes by maintaining beta-cell function. We observed that imatinib counteracted diabetes in two animal models, the streptozotocin-injected mouse and the nonobese diabetes mouse, and that this was paralleled by a partial preservation of the beta-cell mass. In addition, imatinib decreased the death of human beta-cells in vitro when exposed to NO, cytokines, and streptozotocin. The imatinib effect was mimicked by siRNA-mediated knockdown of c-Abl mRNA. Imatinib enhanced beta-cell survival by promoting a state similar to ischemic preconditioning, as evidenced by NF-kappaB activation, increased NO and reactive oxygen species production, and depolarization of the inner mitochondrial membrane. Imatinib did not suppress islet cell death in the presence of an NF-kappaB inhibitor, suggesting that NF kappaB activation is a necessary step in the antiapoptotic action of imatinib. We conclude that imatinib mediates beta-cell survival and that this could contribute to the beneficial effects observed in diabetes. PMID- 17135365 TI - Greatly impaired migration of implanted aquaporin-4-deficient astroglial cells in mouse brain toward a site of injury. AB - We reported previously that astroglia cultured from aquaporin-4-deficient (AQP4-/ ) mice migrate more slowly in vitro than those from wild-type (AQP4+/+) mice (J. Cell Sci. 2005;118, 5691-5698). Here, we investigate the migration of fluorescently labeled AQP4+/+ and AQP4-/- astroglia after implantation into mouse brains in which directional movement was stimulated by a planar stab wound 3 mm away from the axis of the injection needle. Two days after cell injection we determined the location, elongation ratio, and orientation of labeled cells. Migration of AQP4+/+ but not AQP4-/- cells toward the stab was greater than away from the stab. AQP4+/+ astroglia moved on average 1.5 mm toward the stab compared with 0.6 mm for AQP4-/- cells. More than 25% of the migrating AQP4+/+ cells but <3% of AQP4-/- cells appeared elongated (axial ratio>2.5). In transwell assays, AQP4+/+ astroglia migrated faster than AQP4-/- cells in a manner dependent on pore size. At 8 h, approximately 50% of AQP4+/+ cells migrated through 8-microm diameter pores, whereas equivalent migration of AQP4-/- cells was found for 12 microm diameter pores. These results provide in vivo evidence for AQP4-dependent astroglial migration and suggest that modulation of AQP4 expression or function might alter glial scarring. PMID- 17135366 TI - Prohibitin protects against oxidative stress in intestinal epithelial cells. AB - Prohibitin (PHB) is an evolutionarily conserved and ubiquitously expressed protein whose expression or function in intestinal diseases is not known. In this study, we examined the expression and role of PHB in oxidative stress associated with inflammatory bowel disease. Our results show that PHB primarily localizes to the mitochondria in intestinal epithelial cells. Its expression is down-regulated during active human Crohn's disease, experimental colitis in vivo, and oxidative stress in vitro. PHB overexpression increases the expression of glutathione-S transferase pi and protects from oxidant-induced depletion of glutathione. Finally, PHB overexpression decreases accumulation of reactive oxygen metabolites, as well as increased permeability induced by oxidative stress in intestinal epithelial cells. Together, these results suggest that PHB constitutes a previously unrecognized cellular defense against oxidant injury. Thus, strategies to modulate PHB levels may constitute a novel therapeutic approach for intestinal inflammatory diseases, wherein oxidative stress plays a critical role in tissue injury and inflammation. PMID- 17135367 TI - Ribavirin inhibits angiogenesis by tetrahydrobiopterin depletion. AB - Ribavirin is a broad-spectrum antiviral drug that is used to treat hepatitis C virus (HCV)-infected patients. The virological response after ribavirin treatment appears to be insufficient to fully explain ribavirin-induced beneficial effects. Angiogenesis plays a pathogenic role in HCV-induced liver damage. Here, we investigated the influence of therapeutic ribavirin concentrations on angiogenesis. Ribavirin inhibited endothelial cell tube formation in vitro and vessel formation in the chick chorioallantoic membrane assay in vivo. Ribavirin inhibits inosine monophosphate dehydrogenase, which causes depletion of cellular GTP and in turn reduction of cellular tetrahydrobiopterin levels. The availability of tetrahydrobiopterin limits NO production by endothelial NO synthase. Ribavirin reduced levels of tetrahydrobiopterin (as revealed by HPLC), NO (as revealed by electron spin resonance spectroscopy), and cGMP (as revealed by RIA) in endothelial cells. Addition of tetrahydrobiopterin or NO prevented ribavirin-induced tube formation inhibition. In conclusion, angiogenesis inhibition by ribavirin has not been described before. This inhibition may contribute to ribavirin-induced pharmacological effects including adverse events. PMID- 17135368 TI - Vascular expression of germinal ACE fails to maintain normal blood pressure in ACE-/- mice. AB - Maintenance of normal blood pressure is critical for preserving the integrity of the cardiovascular system. Angiotensin 1-converting enzyme (ACE) regulates normal blood pressure and fluid homeostasis through its action in the renin-angiotensin aldosterone system (RAAS) and the renal tubuloglomerular feedback response. Although the two structurally related isozymic forms of ACE both generate the vasoactive octapeptide angiotensin II (Ang II) with equal efficiency, both are expressed in a nonoverlapping tissue-restricted fashion. To discriminate the precise physiological role of each ACE in its requisite tissue in vivo, we expressed one ACE isoform exclusively in a single cell type of an Ace null mouse. Previously, we demonstrated that vascular endothelial cell-specific expression of transgenic somatic ACE (sACE) could restore normal blood pressure of Ace-null mice. In this current study, we expressed germinal ACE (gACE) in the vascular endothelial cells of the Ace null mouse. These mice exhibited correct renal structure, renal function, and normal growth rates. Although the mice had elevated levels of gACE bound to vascular endothelial cells and high levels of gACE and Ang II in the circulating serum, blood pressure was restored only partially. This study demonstrated that gACE, even when expressed in the vasculature, could not functionally substitute for sACE. PMID- 17135369 TI - Voltage-gated K+ channels support proliferation of colonic carcinoma cells. AB - Plasma membrane potassium (K+) channels are required for cell proliferation. Evidence is growing that K+ channels play a central role in the development and growth of human cancer. Here we examine the contribution and the mechanism by which K+ channels control proliferation of T84 human colonic carcinoma cells. Numerous K+ channels are expressed in T84 cells, but only voltage-gated K+ (Kv) channels influenced proliferation. A number of Kv channel inhibitors reduced DNA synthesis and cell number, without exerting apoptotic or toxic effects. Expression of several Kv channels, such as EagI, Kv 3.4 and Kv 1.5, was detected in patch clamp experiments and in fluorescence-based assays using a voltage sensitive dye. The contribution of EagI channels to proliferation was confirmed by siRNA, which abolished EagI activity and inhibited cell growth. Inhibition of Kv channels did not interfere with the ability of T84 cells to regulate their cell vol, but it restricted intracellular pH regulation. In addition, inhibitors of Kv channels, as well as siRNA for EagI, attenuated intracellular Ca2+ signaling. The data suggest that Kv channels control proliferation of colonic cancer cells by affecting intracellular pH and Ca2+ signaling. PMID- 17135370 TI - Retinal functional development is sensitive to environmental enrichment: a role for BDNF. AB - Retina has long been considered less plastic than cortex or hippocampus, the very sites of experience-dependent plasticity. Now, we show that retinal development is responsive to the experience provided by an enriched environment (EE): the maturation of retinal acuity, which is a sensitive index of retinal circuitry development, is strongly accelerated in EE rats. This effect is present also in rats exposed to EE up to P10, that is before eye opening, suggesting that factors sufficient to trigger retinal acuity development are affected by EE during the first days of life. Brain derived neurotrophic factor (BDNF) is precociously expressed in the ganglion cell layer of EE with respect to non-EE rats and reduction of BDNF expression in EE animals counteracts EE effects on retinal acuity. Thus, EE controls the development of retinal circuitry, and this action depends on retinal BDNF expression. PMID- 17135371 TI - Sudden onset panic: epileptic aura or panic disorder? PMID- 17135372 TI - Lessons from neuropsychiatry. PMID- 17135373 TI - The neuropsychiatric profile of Addison's disease: revisiting a forgotten phenomenon. AB - One hundred fifty years since Thomas Addison's original description of the disease, it is not commonly appreciated that patients with Addison's disease may present with psychiatric symptoms. A review of the literature indicates that disturbances in mood, motivation, and behavior are associated with Addison's disease. Psychosis occurs less frequently, but can be the presenting symptom of a life-threatening adrenal crisis. Potential mechanisms for the neuropsychiatric symptoms of Addison's disease include electrophysiological, electrolyte and metabolic abnormalities, glucocorticoid deficiency, increased endorphins, and an associated Hashimoto encephalopathy. Physicians must be aware that Addison's disease may present solely with psychiatric symptoms and maintain a high index of suspicion for this potentially fatal condition. PMID- 17135374 TI - The value of quantitative electroencephalography in clinical psychiatry: a report by the Committee on Research of the American Neuropsychiatric Association. AB - The authors evaluate quantitative electroencephalography (qEEG) as a laboratory test in clinical psychiatry and describe specific techniques, including visual analysis, spectral analysis, univariate comparisons to normative healthy databases, multivariate comparisons to normative healthy and clinical databases, and advanced techniques that hold clinical promise. Controversial aspects of each technique are discussed, as are broader areas of criticism, such as commercial interests and standards of evidence. The published literature is selectively reviewed, and qEEG's applicability is assessed for disorders of childhood (learning and attentional disorders), dementia, mood disorders, anxiety, panic, obsessive-compulsive disorder, and schizophrenia. Emphasis is placed primarily on studies that use qEEG to aid in clinical diagnosis, and secondarily on studies that use qEEG to predict medication response or clinical course. Methodological problems are highlighted, the availability of large databases is discussed, and specific recommendations are made for further research and development. As a clinical laboratory test, qEEG's cautious use is recommended in attentional and learning disabilities of childhood, and in mood and dementing disorders of adulthood. PMID- 17135375 TI - Posttraumatic stress disorder symptoms during the first six months after traumatic brain injury. AB - Controversy exists regarding the rate and risk factors for posttraumatic stress disorder (PTSD) following traumatic brain injury (TBI). The authors determined the rate and phenomenology of PTSD symptoms in the 6 months after TBI by conducting a prospective cohort study of 124 subjects who completed the PTSD Checklist-Civilian Version. The cumulative incidence of meeting PTSD symptom criteria at 6 months was 11% and full criteria 5.6%. Prevalence peaked at 1 month (10%). Eighty-six percent had another psychiatric disorder and 29% a history of PTSD. Symptoms were associated with not completing high school, assault, recalling being terrified or helpless, and positive toxicology. PTSD after TBI is rare and the relation to risk factors and comorbidities must be examined. PMID- 17135376 TI - Aggression and quantitative MRI measures of caudate in patients with chronic schizophrenia or schizoaffective disorder. AB - Caudate dysfunction is implicated in schizophrenia. However, little is known about the relationship between aggression and caudate volumes. Forty-nine patients received magnetic resonance imaging scanning in a double-blind treatment study in which aggression was measured. Caudate volumes were computed using a semiautomated method. The authors measured aggression with the Overt Aggression Scale and the Positive and Negative Syndrome Scale. Larger caudate volumes were associated with greater levels of aggression. The relationship between aggression and caudate volumes may be related to the iatrogenic effects of long-term treatment with typical antipsychotic agents or to a direct effect of schizophrenic processes on the caudate. PMID- 17135377 TI - Awareness of illness in schizophrenia: associations with multiple assessments of executive function. AB - Though insight in schizophrenia is correlated with flexibility in abstract thought, it is unclear how differing dimensions of executive functions are linked to insight. Accordingly, the authors administered the Scale to Assess Unawareness of Mental Disorder and the Delis-Kaplan Executive Function System to 53 participants with schizophrenia spectrum disorders. Spearman Rho correlations revealed that symptom awareness was significantly related to Verbal Fluency, Color-Word, Tower, and Word Context scores. Awareness of treatment need was related to Color-Word, Tower, and Word Context tasks. Results suggest insight may be related to capacities to shift attention between differing environmental demands, plan ahead, and construct contextual understandings. PMID- 17135378 TI - Anxiety and regional cortical glucose metabolism in patients with Alzheimer's disease. AB - In this study, the authors investigated the relationship between anxiety and regional cortical metabolism in Alzheimer's disease. Using the Neuropsychiatric Inventory (NPI), the authors evaluated anxiety in 41 patients with Alzheimer's disease. Regional cortical glucose metabolism was measured using [(18)F] fluorodeoxyglucose positron emission tomography in the resting state. Relationships were assessed using voxel-based (SPM2) and anatomic region-based analyses. Higher NPI anxiety score (frequency x severity) was associated with lower metabolism in bilateral entorhinal cortex, anterior parahippocampal gyrus, and left superior temporal gyrus and insula. Functional activity changes in distinct regions of the cortex contribute to the expression of anxiety in Alzheimer's disease. PMID- 17135379 TI - Delta power in sleep in relation to neuropsychological performance in healthy subjects and schizophrenia patients. AB - Delta power in sleep is of increasing interest because of its association with waking performance in neuropsychological tests. In schizophrenia, this link might be impaired because of a decrease in delta power in sleep and pronounced cognitive deficits. The authors analyzed delta power in sleep and neuropsychological performance in 16 patients with schizophrenia on stable medication with amisulpride and 17 healthy subjects. In healthy subjects, the authors found significant positive correlations between morning performance in declarative memory, procedural learning, and attention and delta power, especially in frontal channels. The authors interpret these results in terms of dysfunctions of thalamocortical and prefrontal networks in schizophrenia. PMID- 17135380 TI - The role of aura in psychopathology and dissociative experiences in epilepsy. AB - Cognitive auras seem to be associated with depression and anxiety, especially in patients with temporal lobe epilepsy (TLE). Dissociative symptoms may occur as an aura or in the context of psychiatric disorders such as depression, anxiety or schizophrenia. This is a cross-sectional study of 62 patients with TLE, using personality and dissociation measures to investigate their relationship with the presence of aura and its different subtypes. Our findings show no difference in psychopathology in patients with different types of aura and reveal that dissociative symptoms correlate with specific measures of anxiety, suggesting a possible link between these experiences and anxiety disorders. PMID- 17135381 TI - Level of cognitive impairment predicts mortality in high-risk community samples: the memory and medical care study. AB - Over the course of 3 years, the authors investigated the relationship between severity of cognitive impairment and mortality in a community sample of 498 elders at high risk for cognitive impairment. Subjects were classified as having no cognitive disorder, mild cognitive impairment, or dementia, based on a validated battery of four neuropsychological tests. Severity of impairment was based on Mini-Mental State Examination scores. Additional data were obtained from subjects' knowledgeable informants and Medicare records. Kaplan-Meier survival estimates and Cox hazard proportion analysis of the sample revealed that presence of cognitive impairment increases mortality in a fashion that parallels the severity of the impairment. PMID- 17135382 TI - A pilot study of quetiapine treatment of aggression due to traumatic brain injury. AB - In a 6-week open-label, flexible dose pilot study of quetiapine for treatment of aggression secondary to traumatic brain injury (TBI), seven subjects who were at least 3-months post-injury were enrolled. The Overt Aggression Scale - Modified (OAS-M) and Clinical Global Impression (CGI) were primary outcome measures. Administration of quetiapine at doses of 25 to 300 mg daily was efficacious and well-tolerated in reducing irritability and aggression resulting from TBI, with an associated improvement in cognitive functioning. PMID- 17135383 TI - Treatment of intractable hiccups with olanzapine following recent severe traumatic brain injury. PMID- 17135384 TI - Persistent amnesia as a sequel of olanzapine-induced neuroleptic malignant syndrome. PMID- 17135385 TI - Nabilone could treat chorea and irritability in Huntington's disease. PMID- 17135386 TI - Essential blepharospasm responding to haloperidol. PMID- 17135387 TI - Using pramipexole in neuropsychiatry: a cautionary note. PMID- 17135388 TI - Interictal psychosis after stroke with forced normalization. PMID- 17135389 TI - Central hyperdopaminergism in peduncular hallucinosis. PMID- 17135390 TI - Othello syndrome secondary to right orbitofrontal lobe excision. PMID- 17135391 TI - Psychiatric manifestation of SSPE. PMID- 17135392 TI - Schizophrenia-like psychosis following right putaminal infarction. PMID- 17135393 TI - Psychogenic movement disorder masquerading as CJD. PMID- 17135394 TI - Effect of organic solvent exposure on chronic kidney disease progression: the GN PROGRESS cohort study. AB - It has been suggested that solvent exposure may have a role in the progression of glomerulonephritis (GN) to ESRD, but this has never been tested with an appropriate cohort study design. A total of 338 non-ESRD patients with a first biopsy for primary GN between 1994 and 2001 were included: 194 IgA nephropathies (IgAN), 75 membranous nephropathies (MN), and 69 FSGS. ESRD, defined as an estimated GFR <15 ml/min per 1.73 m2 or dialysis, was registered during a mean follow-up period of 5 yr. Patients' lifelong solvent exposures before and after diagnosis were recorded by interview and assessed by industrial hygienist experts. Cox models were used to estimate adjusted hazard ratios (HR) of ESRD related to exposures. Overall, 15 and 14% of the patients had been exposed at a low and a high level before diagnosis, respectively. Forty-two with IgAN, 12 with MN, and 22 with FSGS reached ESRD. A graded relationship was observed for MN (age and gender-adjusted HR [95% confidence interval] for low exposure versus none was 3.1 [0.5 to 18.2] and for high exposure versus none was 8.2 [1.9 to 34.7]) and for IgAN (1.6 [0.7 to 3.9] and 2.2 [1.0 to 4.8]) but not for FSGS. Solvent risk was mediated only partly by baseline proteinuria: Adjusted HR for high exposure versus none was 5.5 (1.3 to 23.9) for MN and 1.8 (0.8 to 3.9) for IgAN. In patients with IgAN, there was a trend in increasing HR with exposure duration before and its persistence after diagnosis. These findings support the hypothesized association of solvent exposure with the progression of GN to ESRD. They should prompt clinicians to give greater attention to patients' occupational exposures and possibly to consider professional reclassification. PMID- 17135395 TI - Peroxisome proliferator-activated receptor alpha protects against glomerulonephritis induced by long-term exposure to the plasticizer di-(2 ethylhexyl)phthalate. AB - Safety concerns about di-(2-ethylhexyl)phthalate (DEHP), a plasticizer and a probable endocrine disruptor, have attracted considerable public attention, but there are few studies about long-term exposure to DEHP. DEHP toxicity is thought to involve peroxisome proliferator-activated receptor alpha (PPARalpha), but this contention remains controversial. For investigation of the long-term toxicity of DEHP and determination of whether PPARalpha mediates toxicity, wild-type and PPARalpha-null mice were fed a diet that contained 0.05 or 0.01% DEHP for 22 mo. PPARalpha-null mice that were exposed to DEHP exhibited prominent immune complex glomerulonephritis, most likely related to elevated glomerular oxidative stress. Elevated NADPH oxidase, low antioxidant enzymes, and absence of the PPARalpha dependent anti-inflammatory effects that normally antagonize the NFkappaB signaling pathway accompanied the glomerulonephritis in PPARalpha-null mice. The results reported here indicate that PPARalpha protects against the nephrotoxic effects of long-term exposure to DEHP. PMID- 17135396 TI - Compartmentalization of cAMP-dependent signaling by phosphodiesterase-4D is involved in the regulation of vasopressin-mediated water reabsorption in renal principal cells. AB - The cAMP/protein kinase A (PKA)-dependent insertion of water channel aquaporin-2 (AQP2)-bearing vesicles into the plasma membrane in renal collecting duct principal cells (AQP2 shuttle) constitutes the molecular basis of arginine vasopressin (AVP)-regulated water reabsorption. cAMP/PKA signaling systems are compartmentalized by A kinase anchoring proteins (AKAP) that tether PKA to subcellular sites and by phosphodiesterases (PDE) that terminate PKA signaling through hydrolysis of localized cAMP. In primary cultured principal cells, AVP causes focal activation of PKA. PKA and cAMP-specific phosphodiesterase-4D (PDE4D) are located on AQP2-bearing vesicles. The selective PDE4 inhibitor rolipram increases AKAP-tethered PKA activity on AQP2-bearing vesicles and enhances the AQP2 shuttle and thereby the osmotic water permeability. AKAP18delta, which is located on AQP2-bearing vesicles, directly interacts with PDE4D and PKA. In response to AVP, PDE4D and AQP2 translocate to the plasma membrane. Here PDE4D is activated through PKA phosphorylation and reduces the osmotic water permeability. Taken together, a novel, compartmentalized, and physiologically relevant cAMP-dependent signal transduction module on AQP2 bearing vesicles, comprising anchored PDE4D, AKAP18delta, and PKA, has been identified. PMID- 17135397 TI - Inhibition of histone deacetylase activity suppresses epithelial-to-mesenchymal transition induced by TGF-beta1 in human renal epithelial cells. AB - Histone acetylation plays an important role in regulating gene expressions by modulating chromatin structure. Histone deacetylase (HDAC) inhibitors have been reported to have an antifibrogenic effect in some organs, such as the liver, skin, and lung, but the underlying mechanisms remain to be clarified. In the kidney, bone morphologic protein 7 (BMP-7) and hepatocyte growth factor are reported to antagonize TGF-beta1-induced tubular epithelial-to-mesenchymal transition (EMT), but nothing is known concerning the effect of HDAC inhibitors on EMT. It was shown that trichostatin A (TSA), an HDAC inhibitor, prevented TGF beta1-induced EMT in cultured human renal proximal tubular epithelial cells. Treatment with TGF-beta1 induced morphologic changes such as EMT in human renal proximal tubular epithelial cells. However, co-treatment with TSA completely prevented TGF-beta1-induced morphologic changes and significantly prevented TGF beta1-induced downregulation of E-cadherin and upregulation of collagen type I. Treatment with TSA did not alter TGF-beta1-induced phosphorylation of Smad2 and Smad3 but induced several inhibitory factors of TGF-beta1 signals, such as inhibitors of DNA binding/differentiation 2 (Id2) and BMP-7. Chromatin immunoprecipitation assay confirmed that histone acetylation was involved in the downregulation of E-cadherin and upregulation of Id2 and BMP-7. These results suggest that TSA and other HDAC inhibitors could be new therapeutic agents for tubular EMT. PMID- 17135398 TI - Multidrug resistance-associated protein 4 is involved in the urinary excretion of hydrochlorothiazide and furosemide. AB - The role of ATP-binding cassette transporters in the urinary excretion of diuretics was investigated. Significant ATP-dependent uptake of hydrochlorothiazide (HCT) and furosemide was observed in membrane vesicles that expressed multidrug resistance-associated protein 4 (MRP4) and breast cancer resistance protein (BCRP). Unlike taurocholate uptake, S-methylglutathione had no effect on the ATP-dependent uptake of both compounds by MRP4. The functional importance of MRP4 and BCRP in the urinary excretion of HCT and furosemide was investigated using gene knockout mice. The renal clearance of HCT and furosemide was reduced significantly but not abolished in Mrp4 knockout mice compared with wild-type mice (9.0 +/- 0.9 versus 15 +/- 2 ml/min per kg for HCT and 1.9 +/- 0.3 versus 2.7 +/- 0.1 ml/min per kg for furosemide), and the amount of HCT that was associated with the kidney specimens was greater in Mrp4 knockout mice (21 +/- 3 versus 13 +/- 1 nmol/g kidney). In contrast, Bcrp makes only a negligible contribution because the urinary excretion was unchanged in Bcrp knockout mice. Our results suggest that Mrp4, together with other unknown transporters, accounts for the luminal efflux of HCT and furosemide from proximal tubular epithelial cells. PMID- 17135399 TI - Bone marrow-derived myofibroblasts contribute to the renal interstitial myofibroblast population and produce procollagen I after ischemia/reperfusion in rats. AB - Bone marrow-derived cells (BMDC) have been proposed to exert beneficial effects after renal ischemia/reperfusion injury (IRI) by engraftment in the tubular epithelium. However, BMDC can give rise to myofibroblasts and may contribute to fibrosis. BMDC contribution to the renal interstitial myofibroblast population in relation to fibrotic changes after IRI in rats was investigated. A model of unilateral renal IRI (45 min of ischemia) was used in F344 rats that were reconstituted with R26-human placental alkaline phosphatase transgenic BM to quantify BMDC contribution to the renal interstitial myofibroblast population over time. After IRI, transient increases in collagen III transcription and interstitial protein deposition were observed, peaking on days 7 and 28, respectively. Interstitial infiltrates of BMDC and myofibroblasts reached a maximum on day 7 and gradually decreased afterward. Over time, an average of 32% of all interstitial alpha-smooth muscle actin-positive myofibroblasts coexpressed R26-human placental alkaline phosphatase and, therefore, were derived from the BM. BMD myofibroblasts produced procollagen I protein and therefore were functional. The postischemic kidney environment was profibrotic, as demonstrated by increased transcription of TGF-beta and decreased transcription of bone morphogenic protein-7. TGF-beta protein was present predominantly in interstitial myofibroblasts but not in BMD myofibroblasts. In conclusion, functional BMD myofibroblasts infiltrate in the postischemic renal interstitium and are involved in extracellular matrix production. PMID- 17135400 TI - Rethinking the fear circuit: the central nucleus of the amygdala is required for the acquisition, consolidation, and expression of Pavlovian fear conditioning. AB - In the standard model of pavlovian fear learning, sensory input from neutral and aversive stimuli converge in the lateral nucleus of the amygdala (LA), in which alterations in synaptic transmission encode the association. During fear expression, the LA is thought to engage the central nucleus of the amygdala (CE), which serves as the principal output nucleus for the expression of conditioned fear responses. In the present study, we reexamined the roles of LA and CE. Specifically, we asked whether CE, like LA, might also be involved in fear learning and memory consolidation. Using functional inactivation methods, we first show that CE is involved not only in the expression but also the acquisition of fear conditioning. Next, we show that inhibition of protein synthesis in CE after training impairs fear memory consolidation. These findings indicate that CE is not only involved in fear expression but, like LA, is also involved in the learning and consolidation of pavlovian fear conditioning. PMID- 17135401 TI - CD47 promotes neuronal development through Src- and FRG/Vav2-mediated activation of Rac and Cdc42. AB - The development of axons and dendrites is controlled by small GTP-binding proteins of the Rho family, but the upstream signaling mechanisms responsible for such regulation remain unclear. We have now investigated the role of the transmembrane protein cluster of differentiation 47 (CD47) in this process with hippocampal neurons. CD47-deficient neurons manifested markedly impaired development of dendrites and axons, whereas overexpression of CD47 promoted such development. Interaction of SH2 domain-containing protein tyrosine phosphatase substrate-1 (SHPS-1) with CD47 also induced the formation of dendritic filopodia and spines. These effects of CD47 were prevented by inhibition of either cell division cycle 42 (Cdc42) or Rac. In CD47-deficient neurons, autophosphorylation of Src was markedly reduced. In addition, overexpression of CD47 promoted the autophosphorylation of Src. Inhibition of Src family kinases indeed prevented CD47-promoted dendritic development. Inhibition of either FGD1-related Cdc42 guanine nucleotide exchange factor (GEF) (FRG) or Vav2, which is a GEF for Cdc42 and Rac and is activated by Src, also prevented the effects of CD47 on dendritic development. These results indicate that CD47 promotes development of dendrites and axons in hippocampal neurons in a manner dependent, at least in part, on activation of Cdc42 and Rac mediated by Src as well as by FRG and Vav2. PMID- 17135402 TI - Accelerated accumulation of misfolded prion protein and spongiform degeneration in a Drosophila model of Gerstmann-Straussler-Scheinker syndrome. AB - Prion diseases are CNS disorders that can occur in sporadic, infectious, and inherited forms. Although all forms of prion disease are associated with the accumulation of pathogenic conformers of the prion protein, collectively termed PrP(Sc), the mechanisms by which PrP(Sc) molecules form and cause neuronal degeneration are unknown. Using the bipartite galactosidase-4-upstream activating sequence expression system, we generated transgenic Drosophila melanogaster heterologously expressing either wild-type (WT) or mutant, disease-associated (P101L) mouse PrP molecules in cholinergic neurons. Transgenic flies expressing neuronal P101L PrP molecules exhibited severe locomotor dysfunction and premature death as larvae and adults. These striking clinical abnormalities were accompanied by age-dependent accumulation of misfolded PrP molecules, intracellular PrP aggregates, and neuronal vacuoles. In contrast, transgenic flies expressing comparable levels of WT PrP displayed no clinical, pathological, or biochemical abnormalities. These results indicate that transgenic Drosophila expressing neuronal P101L PrP specifically exhibit several hallmark features of human Gerstmann-Straussler-Scheinker (GSS) syndrome. Because the rates of abnormal PrP accumulation and clinical progression are highly accelerated in Drosophila compared with the rates of these processes in rodents or humans, the P101L mutant may be used for future genetic and pharmacologic studies as a novel invertebrate model of GSS. PMID- 17135403 TI - Hippocampal sharp waves and reactivation during awake states depend on repeated sequential experience. AB - Hippocampal firing patterns during behavior are reactivated during rest and subsequent slow-wave sleep. These reactivations occur during transient local field potential (LFP) events, termed sharp waves. Theories of hippocampal processing suggest that sharp waves arise from strengthened plasticity, and that the strengthened plasticity depends on repeated cofiring of pyramidal cells. We tested these predictions by recording neural ensembles and LFPs from rats running tasks requiring different levels of behavioral repetition. The number of sharp waves emitted increased during sessions with more regular behaviors. Reactivation became more similar to behavioral firing patterns across the session. This enhanced reactivation also depended on the regularity of the behavior. Additional studies in CA3 and CA1 found that the number of sharp waves emitted also increased in CA3 recordings as well as CA1, but that the time courses were different between the two structures. PMID- 17135404 TI - p38 mitogen-activated protein kinase contributes to adenosine A1 receptor mediated synaptic depression in area CA1 of the rat hippocampus. AB - Adenosine is arguably the most potent and widespread presynaptic modulator in the CNS, yet adenosine receptor signal transduction pathways remain unresolved. Here, we demonstrate a novel mechanism in which adenosine A1 receptor stimulation leads to p38 mitogen-activated protein kinase (MAPK) activation and contributes to the inhibition of synaptic transmission. Western blot analysis indicated that selective A1 receptor activation [with N6-cyclopentyladenosine (CPA)] resulted in rapid increases in phosphorylated p38 (phospho-p38) MAPK immunoreactivity in membrane fractions, and decreases in phospho-p38 MAPK in cytosolic fractions. Immunoprecipitation with a phospho-p38 MAPK antibody revealed constitutive association of this phosphoprotein with adenosine A1 receptors. Phospho-p38 MAPK activation by A1 receptor stimulation induced translocation of PP2a (protein phosphatase 2a) to the membrane. We then examined the actions of p38 MAPK activation in A1 receptor-mediated synaptic inhibition. Excitatory postsynaptic field potentials evoked in area CA1 of the rat hippocampus markedly decreased in response to adenosine (10 microM), the A1 receptor agonist CPA (40 nM), or a 5 min exposure to hypoxia. These inhibitory responses were mediated by A1 receptor activation because the selective antagonist DPCPX (8-cyclopentyl-1,3 dipropylxanthine) (100 nM) prevented them. In agreement with the biochemical analysis, the selective p38 MAPK inhibitor SB203580 [4-(4-fluorophenyl)-2-(4 methylsulfinylphenyl)-5-(4-pyridyl)-1H-imidazole] (25 microM) blocked the inhibitory actions of A1 receptor activation, whereas both the inactive analog SB202474 [4-ethyl-2-(p-methoxyphenyl)-5-(4'-pyridyl)-1H-imidazole] (25 microM) and the ERK 1/2 (extracellular signal-regulated kinase 1/2) MAPK inhibitor PD98059 [2'-amino-3'-methoxyflavone] (50 microM) were ineffective. In contrast, the p38 MAPK inhibitors did not inhibit GABA(B)-mediated synaptic depression. These data suggest A1 receptor-mediated p38 MAPK activation is a crucial step underlying the presynaptic inhibitory effect of adenosine on CA3-CA1 synaptic transmission. PMID- 17135405 TI - Human vascular endothelial growth factor protects axotomized retinal ganglion cells in vivo by activating ERK-1/2 and Akt pathways. AB - Based on its trophic effects on neurons and vascular cells, vascular endothelial growth factor (VEGF) is a promising candidate for the treatment of neurodegenerative diseases. To evaluate the therapeutic potential of VEGF, we here examined effects of this growth factor on the degeneration of axotomized retinal ganglion cells (RGCs), which, as CNS-derived neurons, offer themselves in an excellent way to study neuroprotection in vivo. Making use of a transgenic mouse line that constitutively expresses human VEGF under a neuron-specific enolase promoter, we show that (1) the VEGF-transgenic retina overexpresses human VEGF, (2) RGCs carry the VEGF receptor-2, and (3) vascular networks in normal and axotomized VEGF-transgenic (tg) retinas do not differ from control animals. After axotomy, RGCs of VEGF-tg mice were protected against delayed degeneration, as compared with wild-type littermates. Western blots revealed increased phosphorylated ERK-1/2 and Akt and reduced phosphorylated p38 and activated caspase-3 levels in axotomized VEGF-transgenic retinas. Intravitreous injections of pharmacological ERK-1/2 (PD98059) or Akt (LY294002) inhibitors showed that VEGF exerts neuroprotection by dual activation of ERK-1/2 and Akt pathways. In view that axotomy-induced RGC death occurs slowly and considering that RGCs are CNS-derived neurons, we predict the clinical implementation of VEGF in neurodegenerative diseases of both brain and retina. PMID- 17135406 TI - Modular propagation of epileptiform activity: evidence for an inhibitory veto in neocortex. AB - What regulates the spread of activity through cortical circuits? We present here data indicating a pivotal role for a vetoing inhibition restraining modules of pyramidal neurons. We combined fast calcium imaging of network activity with whole-cell recordings to examine epileptiform propagation in mouse neocortical slices. Epileptiform activity was induced by washing Mg2+ ions out of the slice. Pyramidal cells receive barrages of inhibitory inputs in advance of the epileptiform wave. The inhibitory barrages are effectively nullified at low doses of picrotoxin (2.5-5 microM). When present, however, these inhibitory barrages occlude an intense excitatory synaptic drive that would normally exceed action potential threshold by approximately a factor of 10. Despite this level of excitation, the inhibitory barrages suppress firing, thereby limiting further neuronal recruitment to the ictal event. Pyramidal neurons are recruited to the epileptiform event once the inhibitory restraint fails and are recruited in spatially clustered populations (150-250 microm diameter). The recruitment of the cells within a given module is virtually simultaneous, and thus epileptiform events progress in intermittent (0.5-1 Hz) steps across the cortical network. We propose that the interneurons that supply the vetoing inhibition define these modular circuit territories. PMID- 17135407 TI - Insufficient sleep reversibly alters bidirectional synaptic plasticity and NMDA receptor function. AB - Insufficient sleep impairs cognitive functions in humans and animals. However, whether long-term synaptic plasticity, a cellular substrate of learning and memory, is compromised by sleep loss per se remains unclear because of confounding factors related to sleep deprivation (SD) procedures in rodents. Using an ex vivo approach in C57BL/6J mice, we show that sleep loss rapidly and reversibly alters bidirectional synaptic plasticity in the CA1 area of the hippocampus. A brief (approximately 4 h) total SD, respecting the temporal parameters of sleep regulation and maintaining unaltered low corticosterone levels, shifted the modification threshold for long-term depression/long-term potentiation (LTP) along the stimulation frequency axis (1-100 Hz) toward the right. Reducing exposure to sensory stimuli by whisker trimming did not affect the SD-induced changes in synaptic plasticity. Recovery sleep reversed the effects induced by SD. When SD was combined with moderate stress, LTP induction was not only impaired but also occluded. Both electrophysiological analysis and immunoblotting of purified synaptosomes revealed that an alteration in the molecular composition of synaptically activated NMDA receptors toward a greater NR2A/NR2B ratio accompanied the effects of SD. This change was reversed after recovery sleep. By using an unparalleled, particularly mild form of SD, this study describes a novel approach toward dissociating the consequences of insufficient sleep on synaptic plasticity from nonspecific effects accompanying SD in rodents. We establish a framework for cellular models of cognitive impairment related to sleep loss, a major problem in modern society. PMID- 17135408 TI - Disruption of primary motor cortex before learning impairs memory of movement dynamics. AB - Although multiple lines of evidence implicate the primary motor cortex (M1) in motor learning, the precise role of M1 in the adaptation to novel movement dynamics and in the subsequent consolidation of a memory of those dynamics remains unclear. Here we used repetitive transcranial magnetic stimulation (rTMS) to dissociate the contribution of M1 to these distinct aspects of motor learning. Subjects performed reaching movements in velocity-dependent force fields over three epochs: a null-field baseline epoch, a clockwise-field learning epoch (15 min after the baseline epoch), and a clockwise-field retest epoch (24 h after the learning epoch). Half of the subjects received 15 min of 1 Hz rTMS to M1 between the baseline and learning epochs. Subjects given rTMS performed identically to control subjects during the learning epoch. However, control subjects performed with significantly less error than rTMS subjects in the retest epoch on the following day. These results suggest that M1 is not critical to the network supporting motor adaptation per se but that, within this network, M1 may be important for initiating the development of long-term motor memories. PMID- 17135409 TI - Monkey dorsolateral prefrontal cortex sends task-selective signals directly to the superior colliculus. AB - The dorsolateral prefrontal cortex (DLPFC) has been implicated in the ability to perform complex behaviors requiring the implementation of cognitive control. A central supposition of models of prefrontal function is that the DLPFC engages control by selectively modulating the activity of target structures to which it is connected, but no studies in the primate have directly investigated DLPFC output signals. Here, we recorded the activity of DLPFC neurons identified as sending a direct projection to the superior colliculus, a midbrain oculomotor structure, while monkeys performed alternating blocks of trials in which they had to look toward a flashed peripheral stimulus (prosaccades) and trials in which they had to look away from the stimulus in the opposite direction (antisaccades). We report the first direct evidence that the primate DLPFC sends task-selective signals to a target structure. This supports the notion that the DLPFC orchestrates the activity of other brain areas in accordance with task requirements. PMID- 17135410 TI - Directional cuing of target choice in human smooth pursuit eye movements. AB - Perceptual attention and target choice for movement have many features in common. In particular, both generally are based on selection of a particular location in space. To ask whether motor control, like attention, also can exhibit target choice based on nonspatial features of the stimulus, we assessed the initiation of smooth pursuit eye movements when two targets move in different directions after human subjects have been cued which direction or color to track. The direction cue consisted of a patch of dots undergoing either 0% coherent motion or 50% coherent motion in the direction of motion of one of the subsequent targets. After a delay, the fixation spot was extinguished and two spots moved across the same small region of the visual field, one in the cued direction ("target") and one in an orthogonal direction ("distracter"). After the 0% coherent cue, pursuit was approximately the vector average of responses to the two motions presented singly. After the 50% coherent cue, the initial pursuit response was biased strongly toward the target that moved in the cued direction. The impact of the cued direction persisted over delays of up to 1000 ms. Other cues about the direction of upcoming target motion biased the response similarly. Cues about target color also biased pursuit in the direction of motion of the cued target but were considerably less effective than cues indicating the direction of target motion. We conclude that target choice for movement, like perceptual attention, can be based on the features of the chosen target and not only its location in space. PMID- 17135411 TI - Release probability-dependent scaling of the postsynaptic responses at single hippocampal GABAergic synapses. AB - The amount of neurotransmitter released after the arrival of an action potential affects the strength and the trial-to-trial variability of postsynaptic responses. Most studies examining the dependence of synaptic neurotransmitter concentration on the release probability (P(r)) have focused on glutamatergic synapses. Here we asked whether univesicular or multivesicular release characterizes transmission at hippocampal GABAergic synapses. We used multiple probability functional analysis to derive quantal parameters at inhibitory connections between cannabinoid receptor- and cholecystokinin (CCK)-expressing interneurons and CA3 pyramidal cells. After the recordings, the cells were visualized and reconstructed at the light-microscopic level, and the number of boutons mediating the IPSCs was determined using electron microscopy (EM). The number of active zones (AZs) per CCK-immunopositive bouton was determined from three-dimensional EM reconstructions, thus allowing the calculation of the total number of AZs for each pair. Our results reveal an approximate fivefold discrepancy between the numbers of functionally determined release sites (17.4 +/ 3.2) and structurally identified AZs (3.7 +/- 0.9). Channel modeling predicts that a fivefold to sevenfold increase in the peak synaptic GABA concentration is required for the fivefold enhancement of the postsynaptic responses. Kinetic analysis of the unitary IPSCs indicates that the increase in synaptic GABA concentration is most likely attributable to multivesicular release. This change in the synaptic GABA concentration transient together with extremely low postsynaptic receptor occupancy permits a P(r)-dependent scaling of the postsynaptic response generated at a single hippocampal GABAergic synaptic contact. PMID- 17135412 TI - Transplantation of human neural stem cells exerts neuroprotection in a rat model of Parkinson's disease. AB - Neural stem cells (NSCs) possess high potencies of self-renewal and neuronal differentiation. We explored here whether transplantation of human NSCs cloned by v-myc gene transfer, HB1.F3 cells, is a feasible therapeutic option for Parkinson's disease. In vivo, green fluorescent protein-labeled HB1.F3 cells (200,000 viable cells in 3 microl of PBS) when stereotaxically transplanted (same day lesion-transplant paradigm) into the 6-hydroxydopamine-lesioned striatum of rats significantly ameliorated parkinsonian behavioral symptoms compared with controls (vehicle, single bolus, or continuous minipump infusion of trophic factor, or killed cell grafts). Such graft-derived functional effects were accompanied by preservation of tyrosine hydroxylase (TH) immunoreactivity along the nigrostriatal pathway. Grafted HB1.F3 cells survived in the lesioned brain with some labeled with neuronal marker mitogen-activated protein 2 and decorated with synaptophysin-positive terminals. Furthermore, endogenous neurogenesis was activated in the subventricular zone of transplanted rats. To further explore the neuroprotective mechanisms underlying HB1.F3 cell transplantation, we performed cell culture studies and found that a modest number of HB1.F3 cells were TH and dopamine and cAMP-regulated phosphoprotein 32 positive, although most cells were nestin positive, suggesting a mixed population of mature and immature cells. Administration of the HB1.F3 supernatant to human derived dopaminergic SH-SY5Y cells and fetal rat ventral mesencephalic dopaminergic neurons protected against 6-hydroxydopamine neurotoxicity by suppressing apoptosis through Bcl-2 upregulation, which was blocked by anti-stem cell factor antibody alone, the phosphatidylinositol 3-kinase/Akt inhibitor LY294002 [2-(4-morpholinyl)-8-phenyl 1(4H)-benzopyran-4-one] alone, or a combination of both. These results suggest that HB1.F3 cell transplantation exerts neuroprotective effects against dopaminergic depletion in vitro and in vivo because of trophic factor secretion and neuronal differentiation. PMID- 17135413 TI - Contribution of TRPM8 channels to cold transduction in primary sensory neurons and peripheral nerve terminals. AB - Transient receptor potential melastatin 8 (TRPM8) is the best molecular candidate for innocuous cold detection by peripheral thermoreceptor terminals. To dissect out the contribution of this cold- and menthol-gated, nonselective cation channel to cold transduction, we identified BCTC [N-(4-tert-butylphenyl)-4-(3 chloropyridin-2-yl)piperazine-1-carboxamide] as a potent and full blocker of recombinant TRPM8 channels. In cold-sensitive trigeminal ganglion neurons of mice and guinea pig, responses to menthol were abolished by BCTC. In contrast, the effect of BCTC on cold-evoked responses was variable but showed a good correlation with the presence or lack of menthol sensitivity in the same neuron, suggesting a specific blocking action of BCTC on TRPM8 channels. The biophysical properties of native cold-gated currents (I(cold)), and the currents blocked by BCTC were nearly identical, consistent with a role of this channel in cold sensing at the soma. The temperature activation threshold of native TRPM8 channels was significantly warmer than those reported in previous expression studies. The effect of BCTC on native I(cold) was characterized by a dose dependent shift in the temperature threshold of activation. The role of TRPM8 in transduction was further investigated in the guinea pig cornea, a peripheral territory densely innervated with cold thermoreceptors. All cold-sensitive terminals were activated by menthol, suggesting the functional expression of TRPM8 channels in their membrane. However, the spontaneous activity and firing pattern characteristic of cold thermoreceptors was totally immune to TRPM8 channel blockade with BCTC or SKF96365 (1-[2-(4-methoxyphenyl)-2-[3-(4 methoxyphenyl)propoxy]ethyl-1H-imidazole hydrochloride). Cold-evoked responses in corneal terminals were also essentially unaffected by these drugs, whereas responses to menthol were completely abolished. The minor impairment in the ability to transduce cold stimuli by peripheral corneal thermoreceptors during TRPM8 blockade unveils an overlapping functional role for various thermosensitive mechanisms in these nerve terminals. PMID- 17135414 TI - Steady-state adaptation of mechanotransduction modulates the resting potential of auditory hair cells, providing an assay for endolymph [Ca2+]. AB - The auditory hair cell resting potential is critical for proper translation of acoustic signals to the CNS, because it determines their filtering properties, their ability to respond to stimuli of both polarities, and, because the hair cell drives afferent firing rates, the resting potential dictates spontaneous transmitter release. In turtle auditory hair cells, the filtering properties are established by the interactions between BK calcium-activated potassium channels and an L-type calcium channel (electrical resonance). However, both theoretical and in vitro recordings indicate that a third conductance is required to set the resting potential to a point on the I(Ca) and I(BK) activation curves in which filtering is optimized like that found in vivo. Present data elucidate a novel mechanism, likely universal among hair cells, in which mechanoelectric transduction (MET) and its calcium-dependent adaptation provide the depolarizing current to establish the hair cell resting potential. First, mechanical block of the MET current hyperpolarized the membrane potential, resulting in broadband asymmetrical resonance. Second, altering steady-state adaptation by altering the [Ca2+] bathing the hair bundle changed the MET current at rest, the magnitude of which resulted in membrane potential changes that encompassed the best resonant voltage. The Ca2+ sensitivity of adaptation allowed for the first physiological estimate of endolymphatic Ca2+ near the MET channel (56 +/- 11 microM), a value similar to bulk endolymph levels. These effects of MET current on resting potential were independently confirmed using a theoretical model of electrical resonance that included the steady-state MET conductance. PMID- 17135415 TI - Operant conditioning of H-reflex can correct a locomotor abnormality after spinal cord injury in rats. AB - This study asked whether operant conditioning of the H-reflex can modify locomotion in spinal cord-injured rats. Midthoracic transection of the right lateral column of the spinal cord produced a persistent asymmetry in the muscle activity underlying treadmill locomotion. The rats were then either exposed or not exposed to an H-reflex up-conditioning protocol that greatly increased right soleus motoneuron response to primary afferent input, and locomotion was reevaluated. H-reflex up-conditioning increased the right soleus burst and corrected the locomotor asymmetry. In contrast, the locomotor asymmetry persisted in the control rats. These results suggest that appropriately selected reflex conditioning protocols might improve function in people with partial spinal cord injuries. Such protocols might be especially useful when significant regeneration becomes possible and precise methods for reeducating the regenerated spinal cord neurons and synapses are needed for restoring effective function. PMID- 17135416 TI - Neurocytoma is a tumor of adult neuronal progenitor cells. AB - Central neurocytoma (CN) is a rare periventricular tumor, whose derivation, lineage potential, and molecular regulation have been mostly unexplored. We noted that CN cells exhibited an antigenic profile typical of neuronal progenitor cells in vivo, yet in vitro generated neurospheres, divided in response to bFGF (basic fibroblast growth factor), activated the neuroepithelial enhancer of the nestin gene, and gave rise to both neuron-like cells and astrocytes. When CN gene expression was compared with that of both normal adult VZ (ventricular zone) and E/nestin:GFP (green fluorescent protein)-sorted native neuronal progenitors, significant overlap was noted. Marker analysis suggested that the gene expression pattern of CN was that of a proneuronal population; glial markers were conspicuously absent, suggesting that the emergence of astroglia from CN occurred only with passage. The expression pattern of CN was distinguished from that of native progenitor cells by a cohort of differentially expressed genes potentially involved in both the oncogenesis and phenotypic restriction of neurocytoma. These included both IGF2 and several components of its signaling pathway, whose sharp overexpression implicated dysregulated autocrine IGF2 signaling in CN oncogenesis. Both receptors and effectors of canonical wnt signaling, as well as GDF8 (growth differentiation factor 8), PDGF-D, and neuregulin, were differentially overexpressed by CN, suggesting that CN is characterized by the concurrent overactivation of these pathways, which may serve to drive neurocytoma expansion while restricting tumor progenitor phenotype. This strategy of comparing the gene expression of tumor cells to that of the purified native progenitors from which they derive may provide a focused approach to identifying transcripts important to stem and progenitor cell oncogenesis. PMID- 17135417 TI - A gain-of-function mutation in synaptotagmin-1 reveals a critical role of Ca2+ dependent soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex binding in synaptic exocytosis. AB - Synaptotagmin-1, the Ca2+ sensor for fast neurotransmitter release, was proposed to function by Ca2+-dependent phospholipid binding and/or by Ca2+-dependent soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex binding. Extensive in vivo data support the first hypothesis, but testing the second hypothesis has been difficult because no synaptotagmin-1 mutation is known that selectively interferes with SNARE complex binding. Using knock-in mice that carry aspartate-to-asparagine substitutions in a Ca2+-binding site of synaptotagmin-1 (the D232N or D238N substitutions), we now show that the D232N mutation dramatically increases Ca2+-dependent SNARE complex binding by native synaptotagmin-1, but leaves phospholipid binding unchanged. In contrast, the adjacent D238N mutation does not significantly affect SNARE complex binding, but decreases phospholipid binding. Electrophysiological recordings revealed that the D232N mutation increased Ca2+-triggered release, whereas the D238N mutation decreased release. These data establish that fast vesicle exocytosis is driven by a dual Ca2+-dependent activity of synaptotagmin-1, namely Ca2+-dependent binding both to SNARE complexes and to phospholipids. PMID- 17135418 TI - Na(V)1.7 mutant A863P in erythromelalgia: effects of altered activation and steady-state inactivation on excitability of nociceptive dorsal root ganglion neurons. AB - Inherited erythromelalgia/erythermalgia (IEM) is a neuropathy characterized by pain and redness of the extremities that is triggered by warmth. IEM has been associated with missense mutations of the voltage-gated sodium channel Na(V)1.7, which is preferentially expressed in most nociceptive dorsal root ganglia (DRGs) and sympathetic ganglion neurons. Several mutations occur in cytoplasmic linkers of Na(V)1.7, with only two mutations in segment 4 (S4) and S6 of domain I. We report here a simplex case with an alanine 863 substitution by proline (A863P) in S5 of domain II of Na(V)1.7. The functional effect of A863P was investigated by voltage-clamp analysis in human embryonic kidney 293 cells and by current-clamp analysis to determine the effects of A863P on firing properties of small DRG neurons. Activation of mutant channels was shifted by -8 mV, whereas steady-state fast inactivation was shifted by +10 mV, compared with wild-type (WT) channels. There was a marked decrease in the rate of deactivation of mutant channels, and currents elicited by slow ramp depolarizations were 12 times larger than for WT. These results suggested that A863P could render DRG neurons hyperexcitable. We tested this hypothesis by studying properties of rat DRG neurons transfected with either A863P or WT channels. A863P depolarized resting potential of DRG neurons by +6 mV compared with WT channels, reduced the threshold for triggering single action potentials to 63% of that for WT channels, and increased firing frequency of neurons when stimulated with suprathreshold stimuli. Thus, A863P mutant channels produce hyperexcitability in DRG neurons, which contributes to the pathophysiology of IEM. PMID- 17135419 TI - The ducky(2J) mutation in Cacna2d2 results in reduced spontaneous Purkinje cell activity and altered gene expression. AB - The mouse mutant ducky and its allele ducky(2J) represent a model for absence epilepsy characterized by spike-wave seizures and cerebellar ataxia. These mice have mutations in Cacna2d2, which encodes the alpha2delta-2 calcium channel subunit. Of relevance to the ataxic phenotype, alpha2delta-2 mRNA is strongly expressed in cerebellar Purkinje cells (PCs). The Cacna2d2(du2J) mutation results in a 2 bp deletion in the coding region and a complete loss of alpha2delta-2 protein. Here we show that du(2J)/du(2J) mice have a 30% reduction in somatic calcium current and a marked fall in the spontaneous PC firing rate at 22 degrees C, accompanied by a decrease in firing regularity, which is not affected by blocking synaptic input to PCs. At 34 degrees C, du(2J)/du(2J) PCs show no spontaneous intrinsic activity. Du(2J)/du(2J) mice also have alterations in the cerebellar expression of several genes related to PC function. At postnatal day 21, there is an elevation of tyrosine hydroxylase mRNA and a reduction in tenascin-C gene expression. Although du(2J)/+ mice have a marked reduction in alpha2delta-2 protein, they show no fall in PC somatic calcium currents or increase in cerebellar tyrosine hydroxylase gene expression. However, du(2J)/+ PCs do exhibit a significant reduction in firing rate, correlating with the reduction in alpha2delta-2. A hypothesis for future study is that effects on gene expression occur as a result of a reduction in somatic calcium currents, whereas effects on PC firing occur as a long-term result of loss of alpha2delta-2 and/or a reduction in calcium currents and calcium-dependent processes in regions other than the soma. PMID- 17135420 TI - Distinct patterns of striatal medium spiny neuron activity during the natural sleep-wake cycle. AB - Striatal medium-sized spiny neurons (MSNs) integrate and convey information from the cerebral cortex to the output nuclei of the basal ganglia. Intracellular recordings from anesthetized animals show that MSNs undergo spontaneous transitions between hyperpolarized and depolarized states. State transitions, regarded as necessary for eliciting action potential firing in MSNs, are thought to control basal ganglia function by shaping striatal output. Here, we use an anesthetic-free rat preparation to show that the intracellular activity of MSNs is not stereotyped and depends critically on vigilance state. During slow-wave sleep, much as during anesthesia, MSNs displayed rhythmic step-like membrane potential shifts, correlated with cortical field potentials. However, wakefulness was associated with a completely different pattern of temporally disorganized depolarizing synaptic events of variable amplitude. Transitions from slow-wave sleep to wakefulness converted striatal discharge from a cyclic brisk firing to an irregular pattern of action potentials. These findings illuminate different capabilities of information processing in basal ganglia networks, suggesting in particular that a novel style of striatal computation is associated with the waking state. PMID- 17135421 TI - Neural coding of tactile decisions in the human prefrontal cortex. AB - The neural processes underlying tactile decisions in the human brain remain elusive. We addressed this question in a functional magnetic resonance imaging study using a somatosensory discrimination task, requiring participants to compare the frequency of two successive tactile stimuli. Tactile stimuli per se engaged somatosensory, parietal, and frontal cortical regions. Using a statistical model that accounted for the relative difference in frequencies (i.e., Weber fraction) and discrimination accuracy (i.e., correct or incorrect), we show that trial-by-trial relative frequency difference is represented linearly by activity changes in the left dorsolateral prefrontal cortex (DLPFC), the dorsal anterior cingulate cortex, and bilateral anterior insular cortices. However, a circumscribed region within the left DLPFC showed a different response pattern expressed as activity changes that were monotonically related to relative stimulation difference only for correct but not for incorrect trials. Our findings suggest that activity in the left DLPFC encodes stimulus representations that underlie veridical tactile decisions in humans. PMID- 17135422 TI - Rapid activation of inwardly rectifying potassium channels by immobile G-protein coupled receptors. AB - G-protein-coupled receptors (GPCRs) mediate slow synaptic transmission and many other effects of small molecule and peptide neurotransmitters. In the standard model of GPCR signaling, receptors and G-proteins diffuse laterally within the plane of the plasma membrane and encounter each other by random collision. This model predicts that signaling will be most efficient if both GPCRs and G-proteins are free to diffuse, thus maximizing collision frequency. However, neuronal GPCRs are often recruited to and enriched at specific synaptic locations, suggesting receptor mobility is restricted in these cells. Here, we test the hypothesis that restricting GPCR mobility impairs signaling in neurons by limiting the frequency of collisions between receptors and G-proteins. Mu-opioid receptors (MORs) were immobilized on the surface of cerebellar granule neurons by avidin-mediated cross linking, and inwardly rectifying potassium (GIRK) channels were used as rapid indicators of G-protein activation. Mobile and immobile MORs activated GIRK channels with the same onset kinetics and agonist sensitivity in these neurons. In a heterologous expression system, GFP (green fluorescent protein)-tagged G alpha(oA) subunits remained mobile after cross-linking, but their mobility was reduced in the presence of immobile MORs, suggesting that these receptors and subunits were transiently precoupled. In addition, channel activation could be reconstituted with immobile GPCRs, G-protein heterotrimers, and GIRK channels. These results show that collision frequency is not rate-limiting for G-protein activation in CNS neurons, and are consistent with the idea that signaling components are compartmentalized or preassembled. PMID- 17135423 TI - A sex-specific role of type VII adenylyl cyclase in depression. AB - Major depression represents a complex mental disorder. The identification of biological markers that define subtypes of major depressive disorder would greatly facilitate appropriate medical treatments, as well as provide insight into etiology. Reduced activity of the cAMP signaling system has been implicated in the etiology of major depression. Previous work has shown low adenylyl cyclase activity in platelets and postmortem brain tissue of depressed individuals. Here, we investigate the role of the brain type VII isoform of adenylyl cyclase (AC7) in the manifestation of depressive symptoms in genetically modified animals, using a combination of in vivo behavioral experiments, gene expression profiling, and bioinformatics. We also completed studies with humans on the association of polymorphisms in the AC7 gene with major depressive illness (unipolar depression) based on Diagnostic and Statistical Manual of Mental Disorders IV criteria. Collectively, our results demonstrate a sex-specific influence of the AC7 gene on a heritable form of depressive illness. PMID- 17135424 TI - Cell-autonomous beta-catenin signaling regulates cortical precursor proliferation. AB - Overexpression of beta-catenin, a protein that functions in both cell adhesion and signaling, causes expansion of the cerebral cortical precursor population and cortical surface area enlargement. Here, we find that focal elimination of beta catenin from cortical neural precursors in vivo causes premature neuronal differentiation. Precursors within the cerebral cortical ventricular zone exhibit robust beta-catenin-mediated transcriptional activation, which is downregulated as cells exit the ventricular zone. Targeted inhibition of beta-catenin signaling during embryonic development causes cortical precursor cells to prematurely exit the cell cycle, differentiate into neurons, and migrate to the cortical plate. These results show that beta-catenin-mediated transcriptional activation functions in the decision of cortical ventricular zone precursors to proliferate or differentiate during development, and suggest that the cell-autonomous signaling activity of beta-catenin can control the production of cortical neurons and thus regulate cerebral cortical size. PMID- 17135425 TI - Eurados 2006 annual meeting and scientific symposium on uncertainties in dosimetry--principles through to practice. PMID- 17135426 TI - Uncertainties in doses from intakes of radionuclides assessed from monitoring measurements. AB - The evaluation of uncertainties in doses from intakes of radionuclides is one of the most difficult problems in internal dosimetry. In this paper, the process of assessing internal doses from monitoring measurements is reviewed and the major sources of uncertainty are discussed. Methods developed independently at HPA and at IRSN for the determination of uncertainties in internal doses assessed from monitoring measurements are described. Both use a Monte Carlo simulation approach. Results are described for three illustrative examples. An alternative method developed at the Los Alamos National Laboratory that uses Bayesian statistical methods is also described briefly. PMID- 17135427 TI - Relation between concentration of air pollution and cause-specific mortality: four-year exposures to nitrogen dioxide and particulate matter pollutants in 470 neighborhoods in Oslo, Norway. AB - This study investigated the concentration-response relation between air pollution (nitrogen dioxide and particulate matter pollutants PM(10) and PM(2.5)) and cause specific mortality. The population included all inhabitants of Oslo, Norway, aged 51-90 years on January 1, 1992 (n = 143,842) with follow-up of deaths from 1992 to 1998. An air dispersion model (AirQUIS; Norwegian Institute for Air Research (NILU), Oslo, Norway) was used to estimate levels of exposure in 1992-1995 in all 470 administrative neighborhoods. These data were linked to census, education, and death registries. A consistent effect on all causes of death was found for both sexes and age groups by all indicators of air pollution. The effects appeared to increase at nitrogen dioxide levels higher than 40 micro g/m(3) in the youngest age group and with a linear effect in the interval 20-60 micro g/m(3) for the oldest. An effect of all indicators on cardiovascular causes, lung cancer, and chronic obstructive pulmonary disease was also found in both age groups and sexes. The effects were particularly strong for chronic obstructive pulmonary disease, which appeared to have linear effects, whereas cardiovascular causes and lung cancer seemed to have threshold effects. Results show that vulnerable persons with chronic obstructive pulmonary disease and the elderly seem to be susceptible to air pollution at lower levels than the general population. PMID- 17135428 TI - Brain abscess associated with multidrug-resistant Capnocytophaga ochracea infection. AB - Brain abscesses are occasionally associated with a dental source of infection. An unusual case of frontal lobe abscess in a nonimmunocompromised child infected with multidrug-resistant Capnocytophaga ochracea is described and confirms the pathogenic potential of this organism to cause human disease in the central nervous system. PMID- 17135429 TI - Comparison of disk diffusion, VITEK 2, and broth microdilution antimicrobial susceptibility test results for unusual species of Enterobacteriaceae. AB - We compared the antimicrobial susceptibility testing results generated by disk diffusion and the VITEK 2 automated system with the results of the Clinical and Laboratory Standards Institute (CLSI) broth microdilution (BMD) reference method for 61 isolates of unusual species of Enterobacteriaceae. The isolates represented 15 genera and 26 different species, including Buttiauxella, Cedecea, Kluyvera, Leminorella, and Yokenella. Antimicrobial agents included aminoglycosides, carbapenems, cephalosporins, fluoroquinolones, penicillins, and trimethoprim-sulfamethoxazole. CLSI interpretative criteria for Enterobacteriaceae were used. Of the 12 drugs tested by BMD and disk diffusion, 10 showed >95% categorical agreement (CA). CA was lower for ampicillin (80.3%) and cefazolin (77.0%). There were 3 very major errors (all with cefazolin), 1 major error (also with cefazolin), and 26 minor errors. Of the 40 isolates (representing 12 species) that could be identified with the VITEK 2 database, 36 were identified correctly to species level, 1 was identified to genus level only, and 3 were reported as unidentified. VITEK 2 generated MIC results for 42 (68.8%) of 61 isolates, but categorical interpretations (susceptible, intermediate, and resistant) were provided for only 22. For the 17 drugs tested by both BMD and VITEK 2, essential agreement ranged from 80.9 to 100% and CA ranged from 68.2% (ampicillin) to 100%; thirteen drugs exhibited 100% CA. In summary, disk diffusion provides a reliable alternative to BMD for testing of unusual Enterobacteriaceae, some of which cannot be tested, or produce incorrect results, by automated methods. PMID- 17135430 TI - Sequencing of the pncA gene in members of the Mycobacterium tuberculosis complex has important diagnostic applications: Identification of a species-specific pncA mutation in "Mycobacterium canettii" and the reliable and rapid predictor of pyrazinamide resistance. AB - Testing for susceptibility to pyrazinamide (PZA) and analysis of the pncA gene sequences of 423 Mycobacterium tuberculosis complex isolates have revealed a unique silent nucleotide substitution that enables the rapid identification of "M. canettii" (proposed name). Moreover, the lack of a defined mutation within the pncA gene strongly suggests that an alternative mechanism is responsible for PZA resistance. Our results indicate that DNA sequencing of the pncA gene has the potential to shorten the turnaround time and increase the accuracy of PZA susceptibility testing of the M. tuberculosis complex. PMID- 17135431 TI - Identification of sequence diversity in the Escherichia coli fliC genes encoding flagellar types H8 and H40 and its use in typing of Shiga toxin-producing E. coli O8, O22, O111, O174, and O179 strains. AB - Flagellar type H8 is associated with many strains of pathogenic Shiga toxin producing Escherichia coli (STEC), such as O8, O22, O111, O174, and O179 strains. Serological typing of the H8 antigen is limited to motile strains only and suffers from cross-reactivity between flagellar H8 and H40 antigens. In order to develop a method useful for typing of motile and nonmotile STEC O111 and other strains, we have analyzed the flagellar antigen (fliC) genes in representative E. coli H8 and H40 types. Two genotypes of the fliC gene encoding H8 (the fliC-H8 gene) were identified. Genotype fliC-H8a was found to be conserved in STEC O111, O174, and O179 strains; and type fliC-H8b was associated with STEC O8 and O22 strains. Sequence variations were also found in the genetically closely related fliC-H40 gene, although the latter was not found to be associated with STEC strains. A PCR was developed for the specific identification of the fliC-H8 and the fliC-H40 genes in motile and nonmotile E. coli strains. Digestion of PCR products with HhaI resulted in restriction fragment length polymorphisms (RFLPs) which were associated with genotypes fliC-H8a and -H8b as well as with genotypes fliC-H40a and -H40b. The fliC-specific PCR/RFLP typing method was suitable for the rapid typing of motile and nonmotile STEC O8, O22, O111, O174, and O179 strains from different sources whose fliC-H8 genotypes were found to be highly conserved. The fliC genotyping method is advantageous over serotyping and is useful for epidemiological investigations and studies of the evolution of STEC clones. PMID- 17135432 TI - The sensitivity of the OptiMAL rapid diagnostic test to the presence of Plasmodium falciparum gametocytes compromises its ability to monitor treatment outcomes in an area of Papua New Guinea in which malaria is endemic. AB - Using in vivo samples from treatment failure malaria cases, we demonstrate the high sensitivity of the parasite lactase dehydrogenase (pLDH)-based OptiMAL rapid diagnostic test in the detection of P. falciparum gametocytes. This high sensitivity limits the use of pLDH-based tests in the monitoring of treatment outcomes in circumstances where gametocytemia is common. PMID- 17135433 TI - Toward universal varicella-zoster virus (VZV) genotyping: diversity of VZV strains from France and Spain. AB - Thirty-one isolates from France and Spain were genotyped using a published method analyzing DNA sequence variation in open reading frame (ORF) 22, together with an evaluation of three well-characterized single nucleotide polymorphisms (SNP) in ORF 38, 54, and 62. Nineteen were allocated to the European (E) genotype, six were mosaic-1 (M1), and two were mosaic-2 (M2). Four strains were assigned to a new genotype, mosaic-4 (M4). All isolates were wild type, with no Oka vaccine associated markers. No isolates of the mosaic-3 (M3) or Japanese (J) genotype were observed. We also evaluated 13 selected isolates of E, J, M1, and M2 strains (9 of the 31 described above) using an alternative genotyping method based on the assessment of multiple SNP located in ORF 1, 9, 10, 21, 31, 50, 54, 62, and 68. This method assigns wild-type varicella-zoster virus (VZV) strains to seven genotypes: A1, A2, J1, B1, B2, C, and C1. VZV isolates identified as E (ORF22 method) had the genetic signature of genotype C VZV strains, M1 strains were A1, and M2 were A2. No J strains were detected, but parental Oka and vaccine Oka (genotype J) corresponded to genotype J1. M4 isolates (B) share the SNP array observed for M1 and E viruses, and probably represent recombinants between African-Asian (M1) and European (E) viruses. The two genotyping methods, using entirely different genomic targets, produced identical clusters for the strains examined, suggesting robust phylogenetic linkages among VZV strains circulating in Europe. PMID- 17135434 TI - Burkholderia glumae infection in an infant with chronic granulomatous disease. AB - An 8-month-old boy developed a necrotic lung mass from which Burkholderia glumae was recovered, leading to the diagnosis of chronic granulomatous disease (CGD). While other Burkholderia species have been identified as important pathogens in persons with CGD, B. glumae has not been previously reported to cause human infection. PMID- 17135435 TI - Molecular confirmation that Fasciola gigantica can undertake aberrant migrations in human hosts. AB - Two cases of aberrant migration by the liver fluke Fasciola gigantica in humans are reported. In both cases, subadult worms emerged through the skin. The identity of the worms was confirmed from their DNA sequences. This uncommon human pathogen might be more likely than F. hepatica to undertake aberrant migrations in humans. PMID- 17135436 TI - Diversity in Indian equine rotaviruses: identification of genotype G10,P6[1] and G1 strains and a new VP7 genotype (G16) strain in specimens from diarrheic foals in India. AB - Rotaviruses causing severe diarrhea in foals in two organized farms in northern India, during the period from 2003 to 2005, were characterized by electropherotyping, serotyping, and sequence analysis of the genes encoding the outer capsid proteins. Of 137 specimens, 47 (34.31%) were positive for rotavirus and exhibited at least five different electropherotypes (E), E1 to E5. Strains belonging to different electropherotypes exhibited either a different serotype/genotype specificity or a lack of reactivity to typing monoclonal antibodies (MAbs) used in this study. Strains belonging to E1, E2, and E5 exhibited genotype G10,P6[1], G3, and G1 specificities and accounted for 19.0, 42.9, and 9.5% of the isolates, respectively. Though they possessed G10-type VP7, the E1 strains exhibited high reactivity with the G6-specific MAb, suggesting that the uncommon combination of the outer capsid proteins altered the specificity of the conformation-dependent antigenic epitopes on VP7. E3 and E4 strains accounted for 28.6% of the isolates and were untypeable. Sequence analysis of VP7 from E4 strains (Erv92 and Erv99) revealed that they represent a new VP7 genotype, G16. The detection of unexpected bovine rotavirus-derived G10,P6[1] reassortants, G1 serotype strains, and a new genotype (G16) strain in two distant farms reveals an interesting epidemiological situation and diversity of equine rotaviruses in India. PMID- 17135437 TI - Geographic information systems and genotyping in identification of rotavirus G12 infections in residents of an urban slum with subsequent detection in hospitalized children: emergence of G12 genotype in South India. AB - Rotavirus infections by G12 strains in several countries have recently been described. In this study, we report the emergence of G12 strains in south India. Fourteen cases of G12 infection were identified between June and September 2005. G12 was seen in combination with P[6], P[8], or nontypeable P type. Nine cases, including five symptomatic infections and four asymptomatic infections, were identified as part of routine surveillance for rotavirus infections in a birth cohort in the community between June and July 2005. Significant temporal and time space clustering of eight of these cases represents a possible recent introduction of a new rotavirus VP7 genotype. Previous rotavirus infections had been documented for six of the nine children in the community. In the following 2 months, five cases of G12 infection were identified among children presenting to a referral hospital with diarrhea. This is the first description of symptomatic and asymptomatic G12 infections in children in the community. The detection of G12 strains from different parts of the world in recent years suggests the possibility of its emergence as an important global genotype. Monitoring of cocirculating rotavirus strains and detection of emerging strains is important in the context of the availability of rotavirus vaccines. PMID- 17135438 TI - Using a resequencing microarray as a multiple respiratory pathogen detection assay. AB - Simultaneous testing for detection of infectious pathogens that cause similar symptoms (e.g., acute respiratory infections) is invaluable for patient treatment, outbreak prevention, and efficient use of antibiotic and antiviral agents. In addition, such testing may provide information regarding possible coinfections or induced secondary infections, such as virally induced bacterial infections. Furthermore, in many cases, detection of a pathogen requires more than genus/species-level resolution, since harmful agents (e.g., avian influenza virus) are grouped with other, relatively benign common agents, and for every pathogen, finer resolution is useful to allow tracking of the location and nature of mutations leading to strain variations. In this study, a previously developed resequencing microarray that has been demonstrated to have these capabilities was further developed to provide individual detection sensitivity ranging from 10(1) to 10(3) genomic copies for more than 26 respiratory pathogens while still retaining the ability to detect and differentiate between close genetic neighbors. In addition, the study demonstrated that this system allows unambiguous and reproducible sequence-based strain identification of the mixed pathogens. Successful proof-of-concept experiments using clinical specimens show that this approach is potentially very useful for both diagnostics and epidemic surveillance. PMID- 17135439 TI - Development of a flow chart for identification of gram-positive anaerobic cocci in the clinical laboratory. AB - Gram-positive anaerobic cocci (GPAC) are a heterogeneous group of organisms that are isolated from clinical specimens more often than any group of anaerobic bacteria except Bacteroides species, yet many strains are still difficult or impossible to identify in the diagnostic laboratory. In this study, a total of 124 strains, including 13 reference strains of GPAC species and 111 isolates that had been recovered from clinical specimens previously and identified by 16S rRNA gene sequencing, were subjected to biochemical characterization. Based on the results, a short biochemical scheme that involves the minimum essential biochemical tests for accurate identification of clinically important GPAC was developed. PMID- 17135440 TI - Temperate bacteriophages affect pulsed-field gel electrophoresis patterns of Campylobacter jejuni. AB - The recently sequenced genome of Campylobacter jejuni RM1221 revealed the presence of three integrated bacteriophage-like elements. In this study, genes from the first element, a Mu-like bacteriophage, were amplified by PCR and used to probe pulsed-field gels of clinical C. jejuni strains obtained from a waterborne outbreak (Ontario, Canada, 2000). These highly similar strains differed only by their pulsed-field gel electrophoresis (PFGE) patterns due to an apparent insertion or deletion of a 40-kb fragment. Bacteriophage probes hybridized to these different bands in Southern blot analysis, indicating that homologues of bacteriophage genes were present in the outbreak strains. Investigation of the bacteriophage insertion sites in these isolates suggested that bacteriophage acquisition, loss, or transposition was responsible for the PFGE pattern variation. The bacteriophage gene sequences were similar, but not identical, in the outbreak strains and RM1221, indicating that differences may exist between the bacteriophages. PMID- 17135441 TI - Conjugative transfer of chromosomally encoded antibiotic resistance from Helicobacter pylori to Campylobacter jejuni. AB - Many strains of Helicobacter pylori are naturally competent for transformation and able to transfer chromosomal DNA among different isolates using a conjugation like mechanism. In this study, we sought to determine whether H. pylori can transfer DNA into Campylobacter jejuni, a closely related species of the Campylobacterales group. To monitor the transfer, a chromosomally encoded streptomycin resistance cassette prearranged by a specific mutation in the rpsL gene of H. pylori was used. Mating of the bacteria on plates or in liquid broth medium produced C. jejuni progeny containing the streptomycin marker. DNA transfer was unidirectional, from H. pylori to C. jejuni, and the progeny were genetically identical to C. jejuni recipient strains. DNase I treatment reduced but did not eliminate transfer, and DNase I-treated cell supernatants did not transform, ruling out phage transduction. Recombinants also did not occur when the mating bacteria were separated by a membrane, suggesting that DNA transfer requires cell-to-cell contact. Transfer of the streptomycin marker was independent of the H. pylori comB transformation system, the cag pathogenicity island, and another type IV secretion system called tfs3. These findings indicated that a DNase I-resistant, conjugation-like mechanism may contribute to horizontal DNA transfer between different members of the Campylobacteriales group. The significance of this DNA uptake by C. jejuni in the context of acquiring antibiotic resistance is discussed. PMID- 17135442 TI - Discovering potential pathogens among fungi identified as nonsporulating molds. AB - Fungal infections are increasing, particularly among immunocompromised hosts, and a rapid diagnosis is essential to initiate antifungal therapy. Often fungi cannot be identified by conventional methods and are classified as nonsporulating molds (NSM). We sequenced internal transcribed spacer regions from 50 cultures of NSM and found 16 potential pathogens that can be associated with clinical disease. In selected clinical settings, identification of NSM could prove valuable and have an immediate impact on patient management. PMID- 17135443 TI - Characterization of small-colony-variant Stenotrophomonas maltophilia isolated from the sputum specimens of five patients with cystic fibrosis. AB - Cystic fibrosis (CF) patients are predisposed to chronic respiratory infection by nonfermentative gram-negative bacilli, including Stenotrophomonas maltophilia. S. maltophilia is highly resistant to most antibiotics, with the exception of sulfamethoxazole-trimethoprim (SXT). SXT-resistant S. maltophilia has been reported, but the mechanism of resistance is not well defined. Repeated findings of suspected small-colony-variant (SCV) S. maltophilia isolates from the sputa of five CF patients were confirmed by partial 16S rRNA gene sequencing. The SCV S. maltophilia isolates were the only S. maltophilia isolates in these cultures, and none were clonally related. DNA fingerprint analysis confirmed that once established, the SCV S. maltophilia strains persisted. Nutritional studies of SCV S. maltophilia have suggested auxotrophy in hemin, methionine, and thymidine associated with resistance to multiple antibiotics, including SXT. The phenotypic switch from wild-type to SCV S. maltophilia was reproducible in vitro by exposure to SXT, suggesting that prolonged exposure to antibiotics may select for both the SCV S. maltophilia phenotype and SXT resistance by interference with the dihydrofolate reductase pathway. Recovery of SCV S. maltophilia from the sputum of CF patients has implications for both laboratory testing and patient management. PMID- 17135444 TI - Rapid and real-time detection of Chikungunya virus by reverse transcription loop mediated isothermal amplification assay. AB - The standardization and validation of a one-step, single-tube, accelerated, quantitative reverse transcription (RT) loop-mediated isothermal amplification (RT-LAMP) assay targeting the E1 gene for the rapid and real-time detection of Chikungunya virus (CHIKV) are reported. A linear relationship between the amount of template and time of positivity value over a range of 2 x 10(8) to 2 x 10(2) copies was obtained. The feasibility of CHIKV RT-LAMP for clinical diagnosis was validated with patient serum samples from an ongoing epidemic in Southern India. Optimal assay conditions with zero background were established for the detection of low levels of CHIKV in acute-phase patient serum samples. The comparative evaluation of the RT-LAMP assay with acute-phase patient serum samples demonstrated exceptionally higher sensitivity by correctly identifying 21 additional positive borderline cases that were missed by conventional RT-PCR (P < 0.0001) with a detection limit of 20 copies. The quantification of virus load in patient serum samples was also determined from the standard curve based on their time of positivity and was found to be in the range of 2 x 10(8) to 2 x 10(1) copies. In addition, the field applicability of the RT-LAMP assay was also demonstrated by standardizing SYBR Green I-based RT-LAMP wherein the amplification was carried out in a water bath at 63 degrees C for 60 min, which was followed by monitoring gene amplification with the naked eye through color changes. These findings demonstrated that the RT-LAMP assay is a valuable tool for rapid, real-time detection as well as quantification of CHIKV in acute-phase serum samples without requiring any sophisticated equipment and has potential usefulness for clinical diagnosis and surveillance of CHIKV in developing countries. PMID- 17135445 TI - Extreme skewing of annexin II and S100A6 expression identified by proteomic analysis of peritoneal B-1 cells. AB - B-1 cells differ phenotypically, biochemically and functionally from conventional B-2 cells. The origin of these differences remains uncertain. We hypothesized that unbiased analysis of differences in protein expression between B-1 and B-2 cells might provide information not otherwise available, and thus undertook 1 dimensional (1D) gel analysis combined with mass spectrometry. We identified annexin II and S100A6 in peritoneal B-1 cells (B-1P) but not in splenic B-2 cells (B-2S); these results were confirmed by western blot analysis and reflected in mRNA expression. Further analysis of mRNA indicated that elevated expression levels of annexin II and S100A6 were unique to B-1P among several naive lymphoid populations. However, expression of annexin II and S100A6 protein was up regulated in mitogenically stimulated B-2S. In both naive B-1 cells and stimulated B-2 cells, annexin II and S100A6 formed Ca++-sensitive complexes. These results confirm that the emerging field of proteomics detects differentially expressed molecules independently of RNA screening methods. These results identify two proteins (annexin II and S100A6) that are unexpectedly differentially expressed in B-1 cells and, although members of larger families, may fulfill unique, subset-specific functions. These results also validate 1D GE/LC-MS/MS as a reliable screening tool in identifying final protein product expression differences between B-1P and B-2S. PMID- 17135446 TI - Pathological role of Toll-like receptor signaling in cerebral malaria. AB - Toll-like receptors (TLRs) recognize malaria parasites or their metabolites; however, their physiological roles in malaria infection in vivo are not fully understood. Here, we show that myeloid differentiation primary response gene 88 (MyD88)-dependent TLR signaling mediates brain pathogenesis of severe malaria infection, namely cerebral malaria (CM). A significant number of MyD88-, but not TIR domain containing adaptor-inducing IFN-beta (TRIF)-deficient or wild-type (WT) mice survived CM caused by Plasmodium berghei ANKA (PbA) infection. Although systemic parasitemia was comparable, sequestration of parasite and hemozoin load in the brain blood vessels was significantly lower in MyD88-deficient mice compared with those in TRIF-deficient or WT mice. Moreover, brain-specific pathological changes were associated with MyD88-dependent infiltration of CD8+, CCR5+ T cells and CD11c+ dendritic cells, including CD11c+, NK1.1+ and B220+ cells, and up-regulation of genes such as Granzyme B, Lipocalin 2, Ccl3 and Ccr5. Further studies using mice lacking various TLRs suggest that TLR2 and TLR9, but not TLR4, 5 and 7, were involved in CM. These results strongly suggest that TLR2- and/or TLR9-mediated, MyD88-dependent brain pathogenesis may play a critical role in CM, the lethal complication during PbA infection. PMID- 17135447 TI - TGF-beta type I receptor kinase inhibitor down-regulates rheumatoid synoviocytes and prevents the arthritis induced by type II collagen antibody. AB - Rheumatoid arthritis (RA) is characterized by hypertrophic synovial tissues comprising excessively proliferating synovial fibroblasts and infiltrating inflammatory cells. Transforming growth factor-beta (TGF-beta) is a multifunctional cytokine that regulates cell growth, inflammation and angiogenesis by acting on various cell types. In RA synovial tissues, TGF-beta is expressed at high levels. However, the precise role of TGF-beta in RA remains unclear. We herein demonstrated a causal link between the TGF-beta-induced RA synovial cell proliferation and induction of platelet-derived growth factor (PDGF)-AA. In addition, TGF-beta induced IL-6 and vascular endothelial growth factor (VEGF) production by RA synovial fibroblasts associated with nuclear factor-kappa B activation. These effects of TGF-beta on RA synovial fibroblasts were suppressed by TGF-beta type I receptor kinase inhibitor HTS466284. Furthermore, HTS466284 significantly prevented anti-collagen type II antibody induced arthritis in mice according to the clinical manifestations, histology, tumor necrosis factor-alpha, PDGF and VEGF expression and 5-bromo-2'-deoxyuridine incorporation. These in vitro and in vivo results suggest that TGF-beta plays a role in the development of synovial hyperplasia consisting of synovial cell proliferation, inflammation and angiogenesis. The blockade of TGF-beta signaling may thus become an additional strategy for the treatment of RA. PMID- 17135448 TI - Studies in occupational epidemiology and the risk of overadjustment. PMID- 17135449 TI - NO2 and children's respiratory symptoms in the PATY study. AB - OBJECTIVES: NO2)is a major urban air pollutant. Previously reported associations between ambient NO2)and children's respiratory health have been inconsistent, and independent effects of correlated pollutants hard to assess. The authors examined effects of NO2 on a spectrum of 11 respiratory symptoms, controlling for PM10 and SO2, using a large pooled dataset. METHODS: Cross sectional studies were conducted in Russia, Austria, Italy, Switzerland, and the Netherlands, during 1993-99, contributing in total 23 955 children. Study-specific odds ratios for associations with ambient NO2 are estimated using logistic regressions with area level random effects. Heterogeneity between study-specific results, and mean estimates (allowing for heterogeneity) are calculated. RESULTS: Long term average NO2 concentrations were unrelated to prevalences of bronchitis or asthma. Associations were found for sensitivity to inhaled allergens and allergy to pets, with mean odds ratios around 1.14 per 10 microg/m3 NO2. SO2 had little confounding effect, but an initial association between NO2 and morning cough was reduced after controlling for PM10. Associations with reported allergy were not reduced by adjustment for the other pollutants. Odds ratios for allergic symptoms tended to be higher for the 9-12 year old children compared with the 6-8 year old children. CONCLUSIONS: Evidence for associations between NO2 and respiratory symptoms was robust only for inhalation allergies. NO2 most likely is acting as an indicator of traffic related air pollutants, though its direct effect cannot be ruled out. This remains important, as policies to reduce traffic related air pollution will not result in rapid reductions. PMID- 17135450 TI - Monoclonal antibody-based competitive enzyme-linked immunosorbent assay for detecting and quantifying West Nile virus-neutralizing antibodies in horse sera. AB - A rapid immunoassay for detecting and quantifying West Nile virus (WNV) neutralizing antibodies in sera was developed as an alternative to the plaque reduction neutralization test (PRNT), the gold standard test for WNV. The assay is a competitive, enzyme-linked immunosorbent assay using neutralizing monoclonal antibody 5E8 (NT-ELISA). A cutoff percent inhibition (PI) value of 35% (mean PI plus 3 standard deviations), with a specificity of 99%, was established based on analysis of 246 serum samples from horses free of WNV. The NT-ELISA detected neutralizing antibodies in all sera collected 7 or 14 days postinoculation from mice (n = 11) infected with lineage I (strain NY385-99) or II (strain B956) WNV. When sera from WNV-vaccinated horses (n = 212) were tested by NT-ELISA and PRNT, the NT-ELISA gave a positive result for 96.1% (173/180) of the PRNT-positive sera and 3.1% (1/32) of the PRNT-negative sera. Discrepancies between the two tests were observed mainly with sera with low PRNT(90) titers (expressed as the reciprocal of the highest dilution yielding > or = 90% reduction in the number of plaques) for WNV or low PIs by NT-ELISA. The overall agreement (k value) between the two tests was 0.86. A good correlation (r(2) = 0.77) was also observed between the tests for endpoint titration of sera (n = 116). In conclusion, the newly developed NT-ELISA may be a good alternative serologic assay for detecting WNV that can be used for large-scale testing of WNV-neutralizing antibodies in multiple species. PMID- 17135452 TI - Expression of antibodies directed to Paracoccidioides brasiliensis glycosphingolipids during the course of paracoccidioidomycosis treatment. AB - Paracoccidioidomycosis (PCM) is a granulomatous disease caused by the dimorphic fungus Paracoccidioides brasiliensis. The immunoglobulin classes and isotypes of antibodies directed to acidic glycosphingolipids (GSLs) and glucosylceramide of P. brasiliensis were determined by enzyme-linked immunosorbent assay of sera from 31 PCM patients. The reactivities of 38 serum samples were analyzed by considering the stage of treatment: before antifungal treatment (n = 10), during 1 to 4 months of treatment (T1-4; n = 9), during 5 to 12 months of treatment (T5 12; n = 9), and posttreatment (PT; n = 10). Sera from healthy subjects (n = 12) were used as controls. Only the GSL Pb-1 antigen, which presents the carbohydrate structure Galfbeta1-6(Manalpha1-3)Manbeta1, was reactive with the PCM patient sera. The PCM patient sera did not react with Pb-2, which lacks the Galf residue and which is considered the biosynthetic precursor of Pb-1, indicating that the Galf residue is essential for antibody reactivity. The Pb-1 glycolipid from nontreated patients elicited a primary immune response with immunoglobulin M (IgM) production and subsequent switching to IgG1 production. The IgG1 titer increased after the start of antifungal treatment (T1-4 group), and general decreases in the anti-Pb-1 antibody titers were observed after 5 months of treatment (T5-12 and PT groups). The Pb-1 antigen, an acidic GSL with terminal Galf residue, has potential application as an elicitor of the host immune response in patients with PCM. PMID- 17135451 TI - Evaluation of an indirect immunofluorescence assay for strongyloidiasis as a tool for diagnosis and follow-up. AB - The diagnostic accuracy of an indirect immunofluorescence antibody test (IFAT) for Strongyloides stercoralis at different serum antibody titers was evaluated. To assess diagnostic sensitivity, sera from 156 patients with known strongyloidiasis were collected. Negative control sera were obtained from a composite group of 427 subjects (blood donors and hospitalized patients). With an area under the receiver-operating characteristic plot of 0.98, the IFAT showed a high level of diagnostic accuracy for strongyloidiasis. An antibody titer of > or = 1:20, with 97% sensitivity and 98% specificity, was identified as the diagnostic threshold with the best overall performance. Cross-reactions were evaluated with 41 additional samples from patients with other known helminth infections, and the IFAT detected low-titer positivity in only one subject with filariasis. A positive IFAT result at an antibody dilution of > or = 1:80 was virtually 100% specific, with 71% sensitivity. To test the usefulness of the IFAT as a monitoring tool, the changes in specific-antibody titers after treatment in a group of 155 patients were evaluated. Seroreversion or a decrease in antibody titer of twofold or more was observed in 60% of the patients. Response to treatment was directly correlated to the initial antibody titer, and a baseline titer of > or = 1:80 was identified as the best predictor of response. In conclusion, a positive IFAT result at an antibody dilution of >/=1:20 is the optimal cutoff for screening. A titer of > or = 1:80, with virtually no false positive result, is a reliable cutoff for a serological assessment of treatment efficacy and for inclusion in clinical trials. PMID- 17135453 TI - New epidemiological findings on benign paroxysmal positional vertigo. PMID- 17135454 TI - Magnetic resonance perfusion diffusion mismatch, thrombolysis, and clinical outcome in acute stroke. PMID- 17135455 TI - The effect of visual training for patients with visual field defects due to brain damage: a systematic review. AB - The objective of this review was to evaluate whether systematic visual training leads to (1) a restitution of the visual field (restoration), (2) an increase in the visual search field size or an improvement in scanning strategies (compensation) and (3) a transfer of training-related improvements in activities of daily living such as reading. To retrieve relevant publications, computer aided searches of databases (Medline, Embase, Cinahl, Cochrane Central Registers of Controlled Trials) and extensive reference tracing and hand searching were performed. Subsequently, all retrieved and blinded studies were scored on methodological quality. 14 studies were included, 2 randomised controlled trials (RCTs) and 12 within-subject repeated-measures designs (RMD). One of the two RCT studies had good quality. The internal validity of the RMD studies varied from poor to good. Five studies reported a significant effect of the vision restoration therapy (VRT), whereas two studies reported no effect using scanning laser ophthalmoscopy or Goldmann perimetry as outcome measure. All authors of the studies on scanning compensatory therapy (SCT) found a significant effect of up to 30 degrees visual search field, a significant increase in reading speed or decrease in reading errors. It is unclear to what extent patients benefit from restoration therapy in relation to a more efficient scanning strategy which enables them to read faster or to avoid obstacles in a better way. No study has given a satisfactory answer. SCT seems to provide a more successful rehabilitation with more simple and user-friendly training techniques. Validated questionnaires provide the most reliable subjective data to assess the transfer of the relevance of training procedures to activities of daily living of the patient. Hence, SCT is recommended until the effect of the VRT is defined. PMID- 17135458 TI - Self-reported health in people with Parkinson's disease left untreated at diagnosis. PMID- 17135457 TI - Achieved serum magnesium concentrations and occurrence of delayed cerebral ischaemia and poor outcome in aneurysmal subarachnoid haemorrhage. AB - BACKGROUND: Magnesium therapy probably reduces the frequency of delayed cerebral ischaemia (DCI) in subarachnoid haemorrhage (SAH) but uncertainty remains about the optimal serum magnesium concentration. We assessed the relationship between serum magnesium concentrations achieved with magnesium sulphate therapy 64 mmol/day and the occurrence of DCI and poor outcome in patients with SAH. METHODS: Differences in magnesium concentrations between patients with and without DCI and with and without poor outcome were calculated. Quartiles of last serum magnesium concentrations before the onset of DCI, or before the median day of DCI in patients without DCI, were related to the occurrence of DCI and poor outcome at 3 months using logistic regression. RESULTS: Compared with the lowest quartile of serum magnesium concentration (1.10-1.28 mmol/l), the risk of DCI was decreased in each of the higher three quartiles (adjusted odds ratio (OR) in each quartile 0.2; lower 95% CI 0.0 to 0.1; upper limit 0.8 to 0.9). The OR for poor outcome was 1.8 (95% CI 0.5 to 6.9) in the second quartile, 1.0 (95% CI 0.2 to 4.5) in the third quartile and 4.9 (95% CI 1.2 to 19.7) in the highest quartile. DISCUSSION: Magnesium sulphate 64 mmol/day results in a stable risk reduction of DCI over a broad range of achieved serum magnesium concentrations, and strict titration of the dosage therefore does not seem necessary. However, concentrations < or = 1.28 mmol/l could decrease the effect on DCI while concentrations > or = 1.62 might have a negative effect on clinical outcome. PMID- 17135456 TI - Epidemiology of benign paroxysmal positional vertigo: a population based study. AB - OBJECTIVES: To examine the prevalence and incidence, clinical presentation, societal impact and comorbid conditions of benign paroxysmal positional vertigo (BPPV) in the general population. METHODS: Cross-sectional, nationally representative neurotological survey of the general adult population in Germany with a two stage sampling design: screening of 4869 participants from the German National Telephone Health Interview Survey 2003 (response rate 52%) for moderate or severe dizziness or vertigo, followed by validated neurotological interviews (n = 1003; response rate 87%). Diagnostic criteria for BPPV were at least five attacks of vestibular vertigo lasting <1 min without concomitant neurological symptoms and invariably provoked by typical changes in head position. In a concurrent validation study (n = 61) conducted in two specialised dizziness clinics, BPPV was detected by our telephone interview with a specificity of 92% and a sensitivity of 88% (positive predictive value 88%, negative predictive value 92%). RESULTS: BPPV accounted for 8% of individuals with moderate or severe dizziness/vertigo. The lifetime prevalence of BPPV was 2.4%, the 1 year prevalence was 1.6% and the 1 year incidence was 0.6%. The median duration of an episode was 2 weeks. In 86% of affected individuals, BPPV led to medical consultation, interruption of daily activities or sick leave. In total, only 8% of affected participants received effective treatment. On multivariate analysis, age, migraine, hypertension, hyperlipidaemia and stroke were independently associated with BPPV. CONCLUSION: BPPV is a common vestibular disorder leading to significant morbidity, psychosocial impact and medical costs. PMID- 17135459 TI - Neuroimaging findings in human prion disease. AB - Imaging occupies an important role in the investigation of dementia and neurodegenerative disease. The role of imaging in prion disease used to be one of exclusion of other conditions. Over the past decade, the non-invasive nature of MRI, the improved range of magnetic resonance sequences and the availability of clinical and neuropathological correlation have led to a more prominent position of MRI and its inclusion in the diagnostic criteria for variant Creutzfeldt-Jakob disease. As experience of imaging in human prion disease increases, patterns of change related to strain and genotype may improve the diagnostic potential of imaging in the future, may reduce the need for more invasive testing and prove useful in future therapeutic trials. This paper reviews the current knowledge of imaging appearances in human prion disease. PMID- 17135460 TI - Alpha4beta2 nicotinic receptor status in Alzheimer's disease using 123I-5IA-85380 single-photon-emission computed tomography. AB - BACKGROUND: Loss of the alpha4beta2 nicotinic receptor subtype is found at autopsy in Alzheimer's disease. OBJECTIVE: To investigate in vivo changes in this receptor using single-photon-emission CT (SPECT) with 123I-5-iodo-3-[2(S)-2 azetidinylmethoxy] pyridine (5IA-85380), a novel nicotinic acetylcholine receptor ligand which binds predominantly to the alpha4beta2 receptor. METHODS: 32 non smoking subjects (16 with Alzheimer's disease and 16 normal elderly controls) underwent 123I-5IA-85380 and perfusion (99mTc-hexamethylenepropyleneamine oxime (HMPAO)) SPECT scanning. Region of interest analysis was performed with cerebellar normalisation. RESULTS: Significant bilateral reductions in nicotinic receptor binding were identified in frontal (left, p = 0.004; right, p = 0.002), striatal (left, p = 0.004; right, p = 0.003), right medial temporal (p = 0.04) and pons (p<0.001) in patients with AD compared to controls. There were no significant correlations with clinical or cognitive measures. The pattern of nicotinic binding significantly differed from that of perfusion in both patients with AD and controls. Both 123I-5IA-85380 and 99mTc-HMPAO SPECT imaging demonstrated similar diagnostic performance in correctly classifying controls and patients with AD. CONCLUSION: Using 123I-5IA-85380 SPECT we found changes consistent with significant reductions in the nicotinic alpha4beta2 receptor in cortical and striatal brain regions. This method could facilitate diagnosis and may be useful for monitoring progression of the disease and response to treatment in patients with AD and related diseases. PMID- 17135461 TI - Tuatara (Sphenodon) genomics: BAC library construction, sequence survey, and application to the DMRT gene family. AB - The tuatara (Sphenodon punctatus) is of "extraordinary biological interest" as the most distinctive surviving reptilian lineage (Rhyncocephalia) in the world. To provide a genomic resource for an understanding of genome evolution in reptiles, and as part of a larger project to produce genomic resources for various reptiles (evogen.jgi.doe.gov/second_levels/BACs/our_libraries.html), a large-insert bacterial artificial chromosome (BAC) library from a male tuatara was constructed. The library consists of 215 424 individual clones whose average insert size was empirically determined to be 145 kb, yielding a genomic coverage of approximately 6.3x. A BAC-end sequencing analysis of 121 420 bp of sequence revealed a genomic GC content of 46.8%, among the highest observed thus far for vertebrates, and identified several short interspersed repetitive elements (mammalian interspersed repeat-type repeats) and long interspersed repetitive elements, including chicken repeat 1 element. Finally, as a quality control measure the arrayed library was screened with probes corresponding to 2 conserved noncoding regions of the candidate sex-determining gene DMRT1 and the DM domain of the related DMRT2 gene. A deep coverage contig spanning nearly 300 kb was generated, supporting the deep coverage and utility of the library for exploring tuatara genomics. PMID- 17135462 TI - Phylogeography and spatial genetic structure of the Southern torrent salamander: implications for conservation and management. AB - The Southern torrent salamander (Rhyacotriton variegatus) was recently found not warranted for listing under the US Endangered Species Act due to lack of information regarding population fragmentation and gene flow. Found in small order streams associated with late-successional coniferous forests of the US Pacific Northwest, threats to their persistence include disturbance related to timber harvest activities. We conducted a study of genetic diversity throughout this species' range to 1) identify major phylogenetic lineages and phylogeographic barriers and 2) elucidate regional patterns of population genetic and spatial phylogeographic structure. Cytochrome b sequence variation was examined for 189 individuals from 72 localities. We identified 3 major lineages corresponding to nonoverlapping geographic regions: a northern California clade, a central Oregon clade, and a northern Oregon clade. The Yaquina River may be a phylogeographic barrier between the northern Oregon and central Oregon clades, whereas the Smith River in northern California appears to correspond to the discontinuity between the central Oregon and northern California clades. Spatial analyses of genetic variation within regions encompassing major clades indicated that the extent of genetic structure is comparable among regions. We discuss our results in the context of conservation efforts for Southern torrent salamanders. PMID- 17135463 TI - A genomic perspective on the shortcomings of mitochondrial DNA for "barcoding" identification. AB - Approximately 600-bp sequences of mitochondrial DNA (mtDNA) have been designated as "DNA barcodes" and have become one of the most contentious and animated issues in the application of genetic information to global biodiversity assessment and species identification. Advocates of DNA barcodes have received extensive attention and promotion in many popular and refereed scientific publications. However, we suggest that the utility of barcodes is suspect and vulnerable to technical challenges that are particularly pertinent to mtDNA. We review the natural history of mtDNA and discuss problems for barcoding which are particularly associated with mtDNA and inheritance, including reduced effective population size, maternal inheritance, recombination, inconsistent mutation rate, heteroplasmy, and compounding evolutionary processes. The aforementioned could significantly limit the application and utility of mtDNA barcoding efforts. Furthermore, global use of barcodes will require application and acceptance of a barcode-based species concept that has not been evaluated in the context of the extensive literature concerning species designation. Implementation of mtDNA barcodes in spite of technical and practical shortcomings we discuss may degrade the longstanding synthesis of genetic and organism-based research and will not advance studies ranging from genomic evolution to biodiversity assessment. PMID- 17135464 TI - Neural and electromyographic correlates of wrist posture control. AB - In identical experiments in and out of a MR scanner, we recorded functional magnetic resonance imaging and electromyographic correlates of wrist stabilization against constant and time-varying mechanical perturbations. Positioning errors were greatest while stabilizing random torques. Wrist muscle activity lagged changes in joint angular velocity at latencies suggesting trans cortical reflex action. Drift in stabilized hand positions gave rise to frequent, accurately directed, corrective movements, suggesting that the brain maintains separate representations of desired wrist angle for feedback control of posture and the generation of discrete corrections. Two patterns of neural activity were evident in the blood-oxygenation-level-dependent (BOLD) time series obtained during stabilization. A cerebello-thalamo-cortical network showed significant activity whenever position errors were present. Here, changes in activation correlated with moment-by-moment changes in position errors (not force), implicating this network in the feedback control of hand position. A second network, showing elevated activity during stabilization whether errors were present or not, included prefrontal cortex, rostral dorsal premotor and supplementary motor area cortices, and inferior aspects of parietal cortex. BOLD activation in some of these regions correlated with positioning errors integrated over a longer time-frame consistent with optimization of feedback performance via adjustment of the behavioral goal (feedback setpoint) and the planning and execution of internally generated motor actions. The finding that nonoverlapping networks demonstrate differential sensitivity to kinematic performance errors over different time scales supports the hypothesis that in stabilizing the hand, the brain recruits distinct neural systems for feedback control of limb position and for evaluation/adjustment of controller parameters in response to persistent errors. PMID- 17135465 TI - Intrinsically bursting olfactory receptor neurons. AB - Rhythmically bursting neurons are fundamental to neuronal network function but typically are not considered in the context of primary sensory signaling. We now report intrinsically bursting lobster primary olfactory receptor neurons that respond to odors with a phase-dependent burst of action potentials. Rhythmic odor input as might be generated by sniffing entrains the intrinsic bursting rhythm in a concentration-dependent manner and presumably synchronizes the ensemble of bursting cells. We suggest such intrinsically bursting olfactory receptor cells provide a novel way for encoding odor information. PMID- 17135466 TI - Long-term depression in identified stellate neurons of juvenile rat entorhinal cortex. AB - The entorhinal cortex (EC) serves as a gateway to the hippocampus and plays a pivotal role in memory processing in the brain. Superficial layers of the EC convey the cortical input projections to the hippocampus, whereas deep layers of the EC relay hippocampal output projections back to the superficial layers of the EC or to other cortical regions. Whereas the EC expresses long-term potentiation (LTP) and depression (LTD), the underlying cellular and molecular mechanisms have not been determined. Because the axons of the stellate neurons in layer II of the EC form the perforant path that innervates the dentate gyrus granule cells of the hippocampus, we studied the mechanisms underlying the long-term plasticity in identified stellate neurons. Application of high-frequency stimulation (100 Hz for 1 s, repeated 3 times at an interval of 10 s) or forskolin (50 microM) failed to induce significant changes in synaptic strength, whereas application of pairing (presynaptic stimulation at 0.33 Hz paired with postsynaptic depolarization from -60 to -10 mV for 5 min) or low-frequency stimulation (LFS, 1 Hz for 15 min) paradigm-induced LTD. Pairing- or LFS-induced LTDs were N-methyl-D aspartate receptor-dependent and occluded each other suggesting that they have the similar cellular mechanism. Pairing-induced LTD required the activity of calcineurin and involved AMPA receptor endocytosis that required the function of ubiquitin-proteasome system. Our study provides a cellular mechanism that might in part explain the role of the EC in memory. PMID- 17135467 TI - Contextual effects on smooth-pursuit eye movements. AB - Segregating a moving object from its visual context is particularly relevant for the control of smooth-pursuit eye movements. We examined the interaction between a moving object and a stationary or moving visual context to determine the role of the context motion signal in driving pursuit. Eye movements were recorded from human observers to a medium-contrast Gaussian dot that moved horizontally at constant velocity. A peripheral context consisted of two vertically oriented sinusoidal gratings, one above and one below the stimulus trajectory, that were either stationary or drifted into the same or opposite direction as that of the target at different velocities. We found that a stationary context impaired pursuit acceleration and velocity and prolonged pursuit latency. A drifting context enhanced pursuit performance, irrespective of its motion direction. This effect was modulated by context contrast and orientation. When a context was briefly perturbed to move faster or slower eye velocity changed accordingly, but only when the context was drifting along with the target. Perturbing a context into the direction orthogonal to target motion evoked a deviation of the eye opposite to the perturbation direction. We therefore provide evidence for the use of absolute and relative motion cues, or motion assimilation and motion contrast, for the control of smooth-pursuit eye movements. PMID- 17135468 TI - Modulation of serotonergic neurotransmission by nitric oxide. AB - Nitric oxide (NO) and serotonin (5-HT) are two neurotransmitters with important roles in neuromodulation and synaptic plasticity. There is substantial evidence for a morphological and functional overlap between these two neurotransmitter systems, in particular the modulation of 5-HT function by NO. Here we demonstrate for the first time the modulation of an identified serotonergic synapse by NO using the synapse between the cerebral giant cell (CGC) and the B4 neuron within the feeding network of the pond snail Lymnaea stagnalis as a model system. Simultaneous electrophysiological recordings from the pre- and postsynaptic neurons show that blocking endogenous NO production in the intact nervous system significantly reduces the B4 response to CGC activity. The blocking effect is frequency dependent and is strongest at low CGC frequencies. Conversely, bath application of the NO donor DEA/NONOate significantly enhances the CGC-B4 synapse. The modulation of the CGC-B4 synapse is mediated by the soluble guanylate cyclase (sGC)/cGMP pathway as demonstrated by the effects of the sGC antagonist 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ). NO modulation of the CGC-B4 synapse can be mimicked in cell culture, where application of 5-HT puffs to isolated B4 neurons simulates synaptic 5-HT release. Bath application of diethylamine NONOate (DEA/NONOate) enhances the 5-HT induced response in the isolated B4 neuron. However, the cell culture experiment provided no evidence for endogenous NO production in either the CGC or B4 neuron suggesting that NO is produced by an alternative source. Thus we conclude that NO modulates the serotonergic CGC-B4 synapse by enhancing the postsynaptic 5-HT response. PMID- 17135469 TI - Resonance or integration? Self-sustained dynamics and excitability of neural microcircuits. AB - We investigated spontaneous activity and excitability in large networks of artificial spiking neurons. We compared three different spiking neuron models: integrate-and-fire (IF), regular-spiking (RS), and resonator (RES). First, we show that different models have different frequency-dependent response properties, yielding large differences in excitability. Then, we investigate the responsiveness of these models to a single afferent inhibitory/excitatory spike and calibrate the total synaptic drive such that they would exhibit similar peaks of the postsynaptic potentials (PSP). Based on the synaptic calibration, we build large microcircuits of IF, RS, and RES neurons and show that the resonance property favors homeostasis and self-sustainability of the network activity. On the other hand, integration produces instability while it endows the network with other useful properties, such as responsiveness to external inputs. We also investigate other potential sources of stable self-sustained activity and their relation to the membrane properties of neurons. We conclude that resonance and integration at the neuron level might interact in the brain to promote stability as well as flexibility and responsiveness to external input and that membrane properties, in general, are essential for determining the behavior of large networks of neurons. PMID- 17135470 TI - Online maintenance of sensory and motor representations: effects on corticospinal excitability. AB - Flexible behavior requires the ability to delay a response until it is appropriate. This can be achieved by holding either a sensory or a motor representation online. Here we assess whether maintenance of sensory or motor material drives the motor system to different functional states, as indexed by alterations of corticospinal excitability. We used single-pulse TMS to measure corticospinal excitability evoked during the delay period of a novel paradigm in which task contingencies, rather than explicit verbal instructions, induced participants to use either sensory or motor codes to solve a delay-nonmatch-to sample (DNMS) task. This approach allowed us to probe the state of the motor system while the participants were retaining either sensory or motor codes to cross the delay period, rather than the control of short-term storage driven by verbal instructions. When participants could prepare the movement in advance (preparation trials), the excitability of the motor cortex contralateral to the moving hand increased, whereas the excitability of the ipsilateral motor cortex decreased. The increase in excitability was confined to the prime mover, whereas the decrease in excitability extended to cortical territories controlling muscles unrelated to the response. Crucially, these changes in excitability were evoked only during preparation trials and not during trials in which subjects needed to maintain sensory items online (memory trials). We infer that short-term storage of sensory information and preparation of motor responses have differential and specific access to the output stage of the motor system. PMID- 17135471 TI - Warning signals influence motor processing. AB - When observers initiate responses to visual targets, they do so sooner when a preceding stimulus indicates that the target will appear shortly. This consequence of a warning signal may change neural activity in one of four ways. On the sensory side, the warning signal may speed up the rate at which the target is registered by the brain or enhance the magnitude of its signal. On the motor end, the warning signal may lower the threshold required to initiate a response or speed up the rate at which activity accumulates to reach threshold. Here, we describe which explanation is better supported. To accomplish this end, monkeys performed different versions of a cue-target task while we monitored the activity of visuomotor and motor neurons in the superior colliculus. Although the cue target task was designed to measure the properties of reflexive spatial attention, there are two events in this task that produce nonspecific warning effects: a central reorienting event (brightening of central fixation marker) that is used to direct attention away from the cue, and the presentation of the cue itself. Monopolizing on these tendencies, we show that warning effects are associated with several changes in neural activity: the target-related response is enhanced, the threshold for initiating a saccade is lowered, and the rate at which activity accumulates toward threshold rises faster. Ultimately, the accumulation of activity toward threshold predicted behavior most closely. In the discussion, we describe the implications and limitations of these data for theories of warning effects and potential avenues for future research. PMID- 17135472 TI - Intracellular recording reveals temporal integration in inferior colliculus neurons of awake bats. AB - The central nucleus of the inferior colliculus (IC) is a major integrative center in the central auditory system. It receives information from both the ascending and descending auditory pathways. To determine how single IC neurons integrate information over a wide range of sound frequencies and sound levels, we examined their intracellular responses to frequency-modulated (FM) sounds in awake little brown bats (Myotis lucifugus). Postsynaptic potentials were recorded in response to downward FM sweeps of the range typical for little brown bats (80-20 kHz) and to three FM subcomponents (80-60, 60-40, and 40-20 kHz). The majority of recorded neurons responded to the 80- to 20-kHz downward FM sweep with a complex response. In this response an initial hyperpolarization was followed by depolarization with or without spike followed by hyperpolarization. Intracellular recordings in response to three FM subcomponents revealed that these neurons receive excitatory and inhibitory inputs from a wide range of sound frequencies. One third of IC neurons performed nearly linear temporal summation across a wide range of sound frequencies, whereas two thirds of IC neurons exhibited nonlinear summation with different degrees of nonlinearity. Some IC neurons showed different latencies of postsynaptic potentials in response to different FM subcomponents. Often responses to the later FM subcomponent occurred before responses to the earlier ones. This phenomenon may be responsible for response selectivity of IC neurons to FM sweeps. PMID- 17135473 TI - Endogenous mGluR activity suppresses GABAergic transmission in avian cochlear nucleus magnocellularis neurons. AB - GABAergic transmission in the avian cochlear nucleus magnocellularis (NM) of the chick is subject to modulation by gamma-aminobutyric acid type B (GABA(B)) autoreceptors. Here, I investigated modulation of GABAergic transmission in NM by metabotropic glutamate receptors (mGluRs) with whole cell recordings in brain slice preparations. I found that tACPD, a nonspecific mGluR agonist, exerted dose dependent suppression on evoked inhibitory postsynaptic currents (eIPSCs) in NM neurons. At concentrations of 100 or 200 microM, tACPD increased the failure rate of GABAergic transmission. Agonists for group I (3,5-DHPG, 200 microM), group II (DCG-IV, 2 microM), and group III (L-AP4, 10 microM) mGluRs produced a significant reduction in the amplitude of eIPSCs and a significant increase in failure rate, indicating the involvement of multiple mGluRs in this modulation. The frequency, but not the amplitude, of miniature IPSCs (mIPSCs) was decreased significantly by 3,5-DHPG or DCG-IV. Neither frequency nor amplitude of mIPSCs was affected by L-AP4. mGluR antagonists LY341495 (20 microM) plus CPPG (10 microM) significantly increased the amplitude of eIPSCs, indicating that endogenous mGluR activity suppresses GABA release to NM neurons. Furthermore, blockage of mGluRs increased GABA-evoked discharges recorded under physiological Cl(-) concentrations, whereas tACPD (100 microM) eliminated them. The results indicate that mGluRs play important roles in achieving balanced excitation and inhibition in NM and preserving fidelity of temporal information encoded by NM neurons. PMID- 17135474 TI - Gaze-centered updating of remembered visual space during active whole-body translations. AB - Various cortical and sub-cortical brain structures update the gaze-centered coordinates of remembered stimuli to maintain an accurate representation of visual space across eyes rotations and to produce suitable motor plans. A major challenge for the computations by these structures is updating across eye translations. When the eyes translate, objects in front of and behind the eyes' fixation point shift in opposite directions on the retina due to motion parallax. It is not known if the brain uses gaze coordinates to compute parallax in the translational updating of remembered space or if it uses gaze-independent coordinates to maintain spatial constancy across translational motion. We tested this by having subjects view targets, flashed in darkness in front of or behind fixation, then translate their body sideways, and subsequently reach to the memorized target. Reach responses showed parallax-sensitive updating errors: errors increased with depth from fixation and reversed in lateral direction for targets presented at opposite depths from fixation. In a series of control experiments, we ruled out possible biasing factors such as the presence of a fixation light during the translation, the eyes accompanying the hand to the target, and the presence of visual feedback about hand position. Quantitative geometrical analysis confirmed that updating errors were better described by using gaze-centered than gaze-independent coordinates. We conclude that spatial updating for translational motion operates in gaze-centered coordinates. Neural network simulations are presented suggesting that the brain relies on ego velocity signals and stereoscopic depth and direction information in spatial updating during self-motion. PMID- 17135475 TI - Visual FMRI responses in human superior colliculus show a temporal-nasal asymmetry that is absent in lateral geniculate and visual cortex. AB - Eye patching has revealed enhanced saccadic latencies or attention effects when orienting toward visual stimuli presented in the temporal versus nasal hemifields of humans. Such behavioral advantages have been tentatively proposed to reflect possible temporal-nasal differences in the retinotectal pathway to the superior colliculus, rather than in the retinogeniculate pathway or visual cortex. However, this has not been directly tested with physiological measures in humans. Here, we examined responses of the human superior colliculus (SC) to contralateral visual field stimulation, using high spatial resolution fMRI, while manipulating which hemifield was stimulated and orthogonally which eye was patched. The SC responded more strongly to visual stimulation when eye-patching made this stimulation temporal rather than nasal. In contrast, the lateral geniculate nucleus (LGN) plus retinotopic cortical areas V1-V3 did not show any temporal-nasal differences and differed from the SC in this respect. These results provide the first direct physiological demonstration in humans that SC shows temporal-nasal differences that LGN and early visual cortex apparently do not. This may represent a temporal hemifield bias in the strength of the retinotectal pathway, leading to a preference for the contralateral hemifield in the contralateral eye. PMID- 17135476 TI - Combined activation and deactivation of visual cortex during tactile sensory processing. AB - The involvement of occipital cortex in sensory processing is not restricted solely to the visual modality. Tactile processing has been shown to modulate higher-order visual and multisensory integration areas in sighted as well as visually deprived subjects; however, the extent of involvement of early visual cortical areas remains unclear. To investigate this issue, we employed functional magnetic resonance imaging in normally sighted, briefly blindfolded subjects with well-defined visuotopic borders as they tactually explored and rated raised-dot patterns. Tactile task performance resulted in significant activation in primary visual cortex (V1) and deactivation of extrastriate cortical regions V2, V3, V3A, and hV4 with greater deactivation in dorsal subregions and higher visual areas. These results suggest that tactile processing affects occipital cortex via two distinct pathways: a suppressive top-down pathway descending through the visual cortical hierarchy and an excitatory pathway arising from outside the visual cortical hierarchy that drives area V1 directly. PMID- 17135477 TI - Acoustic features of rhesus vocalizations and their representation in the ventrolateral prefrontal cortex. AB - Communication is one of the fundamental components of both human and nonhuman animal behavior. Auditory communication signals (i.e., vocalizations) are especially important in the socioecology of several species of nonhuman primates such as rhesus monkeys. In rhesus, the ventrolateral prefrontal cortex (vPFC) is thought to be part of a circuit involved in representing vocalizations and other auditory objects. To further our understanding of the role of the vPFC in processing vocalizations, we characterized the spectrotemporal features of rhesus vocalizations, compared these features with other classes of natural stimuli, and then related the rhesus-vocalization acoustic features to neural activity. We found that the range of these spectrotemporal features was similar to that found in other ensembles of natural stimuli, including human speech, and identified the subspace of these features that would be particularly informative to discriminate between different vocalizations. In a first neural study, however, we found that the tuning properties of vPFC neurons did not emphasize these particularly informative spectrotemporal features. In a second neural study, we found that a first-order linear model (the spectrotemporal receptive field) is not a good predictor of vPFC activity. The results of these two neural studies are consistent with the hypothesis that the vPFC is not involved in coding the first order acoustic properties of a stimulus but is involved in processing the higher order information needed to form representations of auditory objects. PMID- 17135478 TI - Single-neuron responses to rapidly presented temporal sequences in the primary auditory cortex of the awake macaque monkey. AB - One fundamental process of the auditory system is to process rapidly occurring acoustic stimuli, which are fundamental components of complex stimuli such as animal vocalizations and human speech. Although the auditory cortex is known to subserve the perception of acoustic temporal events, relatively little is currently understood about how single neurons respond to such stimuli. We recorded the responses of single neurons in the primary auditory cortex of alert monkeys performing an auditory task. The stimuli consisted of four tone pips with equal duration and interpip interval, with the first and last pip of the sequence being near the characteristic frequency of the neuron under study. We manipulated the rate of presentation, the frequency of the middle two tone pips, and the order by which they were presented. Our results indicate that single cortical neurons are ineffective at responding to the individual tone pips of the sequence for pip durations of <12 ms, but did begin to respond synchronously to each pip of the sequence at 18-ms durations. In addition, roughly 40% of the neurons tested were able to discriminate the order that the two middle tone pips were presented in at durations of > or =24 ms. These data place the primate primary auditory cortex at an early processing stage of temporal rate discrimination. PMID- 17135479 TI - Frontal eye field contributions to rapid corrective saccades. AB - Visually guided movements can be inaccurate, especially if unexpected events occur while the movement is programmed. Often errors of gaze are corrected before external feedback can be processed. Evidence is presented from macaque monkey frontal eye field (FEF), a cortical area that selects visual targets, allocates attention, and programs saccadic eye movements, for a neural mechanism that can correct saccade errors before visual afferent or performance monitoring signals can register the error. Macaques performed visual search for a color singleton that unpredictably changed position in a circular array as in classic double-step experiments. Consequently, some saccades were directed in error to the original target location. These were followed frequently by unrewarded, corrective saccades to the final target location. We previously showed that visually responsive neurons represent the new target location even if gaze shifted errantly to the original target location. Now we show that the latency of corrective saccades is predicted by the timing of movement-related activity in the FEF. Preceding rapid corrective saccades, the movement-related activity of all neurons began before explicit error signals arise in the medial frontal cortex. The movement-related activity of many neurons began before visual feedback of the error was registered and that of a few neurons began before the error saccade was completed. Thus movement-related activity leading to rapid corrective saccades can be guided by an internal representation of the environment updated with a forward model of the error. PMID- 17135480 TI - Passive soma facilitates submillisecond coincidence detection in the owl's auditory system. AB - Neurons of the avian nucleus laminaris (NL) compute the interaural time difference (ITD) by detecting coincident arrivals of binaural signals with submillisecond accuracy. The cellular mechanisms for this temporal precision have long been studied theoretically and experimentally. The myelinated axon initial segment in the owl's NL neuron and small somatic spikes observed in auditory coincidence detector neurons of various animals suggest that spikes in the NL neuron are generated at the first node of Ranvier and that the soma passively receives back-propagating spikes. To investigate the significance of the "passive soma" structure, we constructed a two-compartment NL neuron model, consisting of a cell body and a first node, and systematically changed the excitability of each compartment. Here, we show that a neuron with a less active soma achieves higher ITD sensitivity and higher noise tolerance with lower energy costs. We also investigate the biophysical mechanism of the computational advantage of the "passive soma" structure by performing sub- and suprathreshold analyses. Setting a spike initiation site with high sodium conductance, not in the large soma but in the small node, serves to amplify high-frequency input signals and to reduce the impact and the energy cost of spike generation. Our results indicate that the owl's NL neuron uses a "passive soma" design for computational and metabolic reasons. PMID- 17135481 TI - Physiological and anatomical evidence for multisensory interactions in auditory cortex. AB - Recent studies, conducted almost exclusively in primates, have shown that several cortical areas usually associated with modality-specific sensory processing are subject to influences from other senses. Here we demonstrate using single-unit recordings and estimates of mutual information that visual stimuli can influence the activity of units in the auditory cortex of anesthetized ferrets. In many cases, these units were also acoustically responsive and frequently transmitted more information in their spike discharge patterns in response to paired visual auditory stimulation than when either modality was presented by itself. For each stimulus, this information was conveyed by a combination of spike count and spike timing. Even in primary auditory areas (primary auditory cortex [A1] and anterior auditory field [AAF]), approximately 15% of recorded units were found to have nonauditory input. This proportion increased in the higher level fields that lie ventral to A1/AAF and was highest in the anterior ventral field, where nearly 50% of the units were found to be responsive to visual stimuli only and a further quarter to both visual and auditory stimuli. Within each field, the pure-tone response properties of neurons sensitive to visual stimuli did not differ in any systematic way from those of visually unresponsive neurons. Neural tracer injections revealed direct inputs from visual cortex into auditory cortex, indicating a potential source of origin for the visual responses. Primary visual cortex projects sparsely to A1, whereas higher visual areas innervate auditory areas in a field-specific manner. These data indicate that multisensory convergence and integration are features common to all auditory cortical areas but are especially prevalent in higher areas. PMID- 17135483 TI - Differential expression of glucose transporters in rabbit placenta: effect of hypercholesterolemia in dams. AB - Low birth weight is observed in rabbit offspring when maternal hypercholesterolemia is induced during gestation, but the related etiology is still unknown. Glucose is one of the most important substances during fetal development, and defect in glucose supply to fetus was related to pathophysiological mechanisms in intrauterine growth restriction. Thus, the aim of this work was to evaluate the impact of maternal hypercholesterolemia during rabbit gestation on the glucose metabolism and the routing of glucose transporters (SLC2 and SLC5 [previously known as GLUT and SGLT]) in placenta. In this study, maternal and offspring serum levels of glucose and insulin were evaluated for control and hypercholesterolemic groups, and the mRNA and protein expressions of placental SLCs were quantified by real-time RT-PCR and Western immunoblot, respectively. Our data demonstrate that maternal hypercholesterolemia during gestation: 1) induces offspring hypoglycemia; 2) does not modify the genetic and protein expressions of SLC2A1 and SLC2A4 (previously GLUT1 and GLUT4) in total placental extract; 3) downregulates the placental SLC5A1 (previously SGLT1) protein expression without affecting its mRNA levels; 4) impairs the translocation of SLC2A1 but not SLC2A4 from cytoplasmatic pool to the cell membrane surface. Then we assume that reduction of offspring birth weight in presence of maternal hypercholesterolemia may be related to the offspring's hypoglycemia and the reduction of the cell surface expression of placental SLC2A1. PMID- 17135484 TI - The position of yeast snoRNA-coding regions within host introns is essential for their biosynthesis and for efficient splicing of the host pre-mRNA. AB - Genomic location of sequences encoding small nucleolar RNAs (snoRNAs) is peculiar in all eukaryotes from yeast to mammals: most of them are encoded within the introns of host genes. In Saccharomyces cerevisiae, seven snoRNAs show this location. In this work we demonstrate that the position of snoRNA-coding regions with respect to splicing consensus sequences is critical: yeast strains expressing mutant constructs containing shorter or longer spacers (the regions between snoRNA ends and intron splice sites) show a drop in accumulation of U24 and U18 snoRNAs. Further mutational analysis demonstrates that altering the distance between the 3' end of the snoRNA and the branch point is the most important constraint for snoRNA biosynthesis, and that stable external stems, which are sometimes present in introns containing snoRNAs, can overcome the positional effect. Surprisingly enough, splicing of the host introns is clearly affected in most of these constructs indicating that, at least in S. cerevisiae, an incorrect location of snoRNA-coding sequences within the host intron is detrimental to the splicing process. This is different with respect to what was demonstrated in mammals, where the activity of the splicing machinery seems to be dominant with respect to the assembly of snoRNPs, and it is not affected by the location of snoRNA sequences. We also show that intronic box C/D snoRNA recognition and assembly of snoRNPs occur during transcription when splicing sequences are recognized. PMID- 17135482 TI - Neurophysiological endophenotypes of schizophrenia: the viability of selected candidate measures. AB - In an effort to reveal susceptibility genes, schizophrenia research has turned to the endophenotype strategy. Endophenotypes are characteristics that reflect the actions of genes predisposing an individual to a disorder, even in the absence of diagnosable pathology. Individual endophenotypes are presumably determined by fewer genes than the more complex phenotype of schizophrenia and would, therefore, reduce the complexity of genetic analyses. Unfortunately, despite there being rational criteria to define a viable endophenotype, the term is sometimes applied indiscriminately to characteristics that are deviant in affected individuals. Schizophrenia patients exhibit deficits in several neurophysiological measures of information processing that have been proposed as candidate endophenotypes. Successful processing of sensory inputs requires the ability to inhibit intrinsic responses to redundant stimuli and, reciprocally, to facilitate responses to less frequent salient stimuli. There is evidence to suggest that both these processes are "impaired" in schizophrenia. Measures of inhibitory failure include prepulse inhibition of the startle reflex, P50 auditory evoked potential suppression, and antisaccade eye movements. Measures of impaired deviance detection include mismatch negativity and the P300 event related potential. The purpose of this review is to systematically evaluate the endophenotype candidacy of these key neurophysiological abilities. For each candidate, we describe typical experimental procedures, the current understanding of the underlying neurobiology, the nature of the abnormality in schizophrenia, the reliability, stability and heritability of the measure, and any reported gene associations. We conclude with a discussion of the few studies thus far that have employed a multivariate approach with these candidates. PMID- 17135485 TI - Dynamic association and localization of human H/ACA RNP proteins. AB - Mammalian H/ACA RNPs are essential for ribosome biogenesis, pre-mRNA splicing, and telomere maintenance. To form mature RNA-protein complexes, one H/ACA RNA associates with four core proteins. In the cell, this process is assisted by at least one nuclear assembly factor, NAF1. Here we report several unanticipated dynamic aspects of H/ACA RNP proteins. First, when overexpressed, NAF1 delocalizes to the cytoplasm. However, its nucleocytoplasmic shuttling properties remain unaffected. These observations demonstrate a subtle equilibrium between NAF1 expression levels and the availability of NAF1 nuclear binding sites. Second, although NAF1 is excluded from mature RNPs in nucleoli and Cajal bodies, NAF1 associates with mature H/ACA RNA in cell lysates. This association occurs post-lysis because it is observed even when NAF1 and the H/ACA RNA are expressed in separate cells. This documents a protein-RNP association in cell lysates that is absent from intact cells. Third, in similar experiments, all H/ACA core proteins, except NAP57, exchange with their exogenous counterparts, portraying an unexpected dynamic picture of H/ACA RNPs. Finally, the irreversible association of only NAP57 with H/ACA RNA and the conundrum that only NAP57 is mutated in X linked dyskeratosis congenita (even though most core proteins are required for maintaining H/ACA RNAs) may be more than a coincidence. PMID- 17135486 TI - Characterization of the short RNAs bound by the P19 suppressor of RNA silencing in mouse embryonic stem cells. AB - Studies of mammalian RNA interference (RNAi) have focused largely on the actions of microRNAs; however, in other organisms, endogenous short-interfering RNAs (siRNAs) are involved in silencing processes. To date, similar molecules have been difficult to characterize in mammalian cells. P19 is a plant suppressor of RNA silencing that binds with high affinity to siRNAs. Here, the short RNAs bound by P19 in mouse embryonic stem (ES) cells have been characterized. We show that P19 selectively immunoprecipitates endogenous short RNAs from ES cells. Cloning of immunoprecipitated RNA reveals a strong selection for short RNAs that are exact matches to ribosomal RNA (rRNA), with particular short rRNA species highly enriched in P19 immunoprecipitates. Complementary strands to the enriched rRNAs were not cloned, which was surprising because P19 was previously thought to bind only siRNAs. We show that P19 binds tightly to a noncanonical dsRNA substrate comprised of a short RNA annealed to a much longer partner, such that the double stranded region between the two is 19 base pairs long. Binding to similar endogenous species might explain the association of P19 with short rRNAs in ES cells. Finally, we show that the P19-enriched rRNAs are not involved in canonical RNAi, as they exist in the absence of Dicer and do not function as post transcriptional gene silencers. Our results support the previous observation that endogenous siRNAs are not abundant molecules in mouse ES cells. PMID- 17135487 TI - Genetic and biochemical characterization of Drosophila Snipper: A promiscuous member of the metazoan 3'hExo/ERI-1 family of 3' to 5' exonucleases. AB - The DnaQ-H family exonuclease Snipper (Snp) is a 33-kDa Drosophila melanogaster homolog of 3'hExo and ERI-1, exoribonucleases implicated in the degradation of histone mRNA in mammals and in the negative regulation of RNA interference (RNAi) in Caenorhabditis elegans, respectively. In metazoans, Snp, Exod1, 3'hExo, ERI-1, and the prpip nucleases define a new subclass of structure-specific 3'-5' exonucleases that bind and degrade double-stranded RNA and/or DNA substrates with 3' overhangs of 2-5 nucleotides (nt) in the presence of Mg2+ with no apparent sequence specificity. These nucleases are also capable of degrading linear substrates. Snp efficiently degrades structured RNA and DNA substrates as long as there exists a minimum 3' overhang of 2 nt to initiate degradation. We identified a Snp mutant and used it to test whether Snp plays a role in regulating histone mRNA degradation or RNAi in vivo. Snp mutant flies are viable, and display no obvious developmental abnormalities. The expression pattern and level of histone H3 mRNA in Snp mutant embryos and third instar imaginal eye discs was indistinguishable from wild type, suggesting that Snp does not play a significant role in the turnover of histone mRNA at the end of the S phase. The loss of Snp was also unable to enhance the silencing capability of two different RNAi transgenes targeting the white and yellow genes, suggesting that Snp does not negatively modulate RNAi. Therefore, Snp is a nonessential exonuclease that is not a functional ortholog of either 3'hExo or ERI-1. PMID- 17135488 TI - The precursor tRNA 3'-CCA interaction with Escherichia coli RNase P RNA is essential for catalysis by RNase P in vivo. AB - The L15 region of Escherichia coli RNase P RNA forms two Watson-Crick base pairs with precursor tRNA 3'-CCA termini (G292-C75 and G293-C74). Here, we analyzed the phenotypes associated with disruption of the G292-C75 or G293-C74 pair in vivo. Mutant RNase P RNA alleles (rnpBC292 and rnpBC293) caused severe growth defects in the E. coli rnpB mutant strain DW2 and abolished growth in the newly constructed mutant strain BW, in which chromosomal rnpB expression strictly depended on the presence of arabinose. An isosteric C293-G74 base pair, but not a C292-G75 pair, fully restored catalytic performance in vivo, as shown for processing of precursor 4.5S RNA. This demonstrates that the base identity of G292, but not G293, contributes to the catalytic process in vivo. Activity assays with mutant RNase P holoenzymes assembled in vivo or in vitro revealed that the C292/293 mutations cause a severe functional defect at low Mg2+ concentrations (2 mM), which we infer to be on the level of catalytically important Mg2+ recruitment. At 4.5 mM Mg2+, activity of mutant relative to the wild-type holoenzyme, was decreased only about twofold, but 13- to 24-fold at 2 mM Mg2+. Moreover, our findings make it unlikely that the C292/293 phenotypes include significant contributions from defects in protein binding, substrate affinity, or RNA degradation. However, native PAGE experiments revealed nonidentical RNA folding equilibria for the wild-type versus mutant RNase P RNAs, in a buffer- and preincubation-dependent manner. Thus, we cannot exclude that altered folding of the mutant RNAs may have also contributed to their in vivo defect. PMID- 17135489 TI - Self-splicing of a group I intron reveals partitioning of native and misfolded RNA populations in yeast. AB - Stable RNAs must form specific three-dimensional structures, yet many RNAs become kinetically trapped in misfolded conformations. To understand the factors that control the accuracy of RNA folding in the cell, the self-splicing activity of the Tetrahymena group I intron was compared in different genetic contexts in budding yeast. The extent of splicing was 98% when the intron was placed in its natural rDNA context, but only 3% when the intron was expressed in an exogenous pre-mRNA. Further experiments showed that the probability of forming the active intron structure depends on local sequence context and transcription by Pol I. Pre-rRNAs decayed at similar rates, whether the intron was wild type or inactivated by an internal deletion, suggesting that most of the unreacted pre rRNA is incompetent to splice. Northern blots and complementation assays showed that mutations that destabilize the intron tertiary structure inhibited self splicing and processing of internal transcribed spacer 2. The data are consistent with partitioning of pre-rRNAs into active and inactive populations. The misfolded RNAs are sequestered and degraded without refolding to a significant extent. Thus, the initial fidelity of folding can dictate the intracellular fate of transcripts containing this group I intron. PMID- 17135490 TI - Biodiversity and emerging diseases. AB - First we remind general considerations concerning biodiversity on earth and particularly the loss of genetic biodiversity that seems irreversible whether its origin is directly or indirectly linked to human activities. Urgent and considerable efforts must be made from now on to cataloge, understand, preserve, and enhance the value of biodiversity while ensuring food safety and human and animal health. Ambitious integrated and multifield research programs must be implemented in order to understand the causes and anticipate the consequences of loss of biodiversity. Such losses are a serious threat to sustainable development and to the quality of life of future generations. They have an influence on the natural balance of global biodiversity in particularly in reducing the capability of species to adapt rapidly by genetic mutations to survive in modified ecosystems. Usually, the natural immune systems of mammals (both human and animal), are highly polymorphic and able to adapt rapidly to new situations. We more specifically discuss the fact that if the genetic diversity of the affected populations is low the invading microorganisms, will suddenly expand and create epidemic outbreaks with risks of pandemic. So biodiversity appears to function as an important barrier (buffer), especially against disease-causing organisms, which can function in different ways. Finally, we discuss the importance of preserving biodiversity mainly in the wildlife ecosystems as an integrated and sustainable approach among others in order to prevent and control the emergence or reemergence of diseases in animals and humans (zoonosis). Although plants are also part of this paradigm, they fall outside our field of study. PMID- 17135491 TI - Perspectives on applied spatial analysis to animal health: a case of rodents in Thailand. AB - Geographic information systems (GIS) and remote sensing have been increasingly used in ecology and epidemiology, providing a spatial approach for animal health issues. Recent development of earth environmental satellites--i.e., their growing number, improving sensor resolutions and capabilities--has offered new opportunities to delineate possible habitats and understand animals and associated parasites in their environment, by identifying the nature and structure of land use, hydrological network, soil hydromorphy, and human settlements. Integrated into GIS, remotely sensed and other geo-referenced data allow both spatial and temporal analyses of animal ecology and health. However, a review of their applications has showed the poor quality of data sources and processing used, revealing limitations between theory and practical implementations. As an example, the assessment of the expected distribution of Bandicoot rats, main agricultural pest and vector of zoonoses in Phrae province (North Thailand), illustrates a rational use of spatial analysis, with the choice of relevant data, scales, and processing. Vegetation indices are computed on a TERRA ASTER image and further classified using elevation data. The biotopes of Bandicota indica and Bandicota savilei are delimited, providing a major source of knowledge for rodent and human health analyses. PMID- 17135492 TI - Epidemics of emerging animal diseases and food-borne infection problems over the last 5 years in Japan. AB - There have been several emerging animal diseases and food-borne infection problems occurring in Japan over the last 5 years. We describe brief pictures of these epidemics and our control activities. As acute contagious and/or emerging animal diseases, the foot and mouth disease (FMD) outbreak caused by the Pan Asian topotype of the type O virus occurred in March 2000 after 92 years of FMD free status. In 2004, four cases of the highly pathogenic avian influenza (HPAI), which was the first outbreak after 79 years, and caused by the H5N1 subtype, were identified. As part of the responses against these outbreaks, all the animals in the affected farms were destroyed, and movement control areas were established around the infected premises, and a nation-wide intensive survey for FMD and HPAI was performed. As for food-borne or feed-borne infections, the first bovine spongiform encephalopathy (BSE) was identified in September 2001 and 19 more cases have been reported until June 2005. A large outbreak of food-borne infection caused by low-fat milk contaminated with enterotoxin A produced by Staphylococcus aureus, involving more than 13,000 patients, occurred in 2000. In 2003, people who consumed uncooked liver and meat from wild boar and deer developed clinical signs of hepatitis caused by the hepatitis E virus. Pork is also suspected as natural source of virus transmission. Early detection of the first cases and rapid action in preventing and controlling the spread of infections are very important combined with proper risk communication about correct information of the diseases. PMID- 17135493 TI - Implication of phylogenetic systematics of rodent-borne hantaviruses allows understanding of their distribution. AB - Hantaviruses' distribution is reassessed after performing a cladistic analysis on 93 strains isolated from rodents, and one used as outgroup: Thottapalayam isolated from a shrew. While most hantaviruses found in wild animals were collected in northern Asia, Europe, North America, and South America, only Thottapalayam and Thailand were found in South and Southeastern Asia. Thottapalayam is highly divergent from the other known hantaviruses and may represent the emerging tip of a different lineage. Serological surveys carried out to detect evidence of Hantavirus in human populations revealed positive samples not only in West and Central Africa but also in Thailand, with a first case recently confirmed. This suggests that Hantaan-related viruses may infect humans out of their well-documented range. Thus, if rodents are probably the primary reservoir, other mammals may be involved in the cycle of hantaviruses. Additional work is needed out of the traditional areas where hantaviruses have been recorded. New viruses, different hosts, and different human syndromes may be discovered in the future mainly in Southeastern Asia and in Africa where Muridae rodents are present and highly diversified. PMID- 17135494 TI - Epizootics of yellow fever in Venezuela (2004-2005): an emerging zoonotic disease. AB - Epidemics and epizootics of yellow fever (YF) have been occurring in the border area of eastern Colombia and western Venezuela since 2003; for this reason many epidemiological control measures were adopted by the Ministry of Health (MOH) trying to prevent their spreading. These activities included monkey deaths surveillance as well as immunization of susceptible individuals with YF vaccine. In this setting, we analyzed epidemiological and epizootical issues related to YF in Venezuela during 2004-2005. In this period, YF epizootics occurred initially without geographical links to the 2003 outbreaks (which occurred at the Southern Maracaibo lake epizootic wave), but in relation with the Guayana epizootic wave; beginning in Monagas state and then affecting Anzoategui, Guarico, and Sucre states. Just months later, Apure was also affected. Merida and Tachira also report epizootics for the end of 2004. This year concluded with 15 human deaths due to YF and more than 100 howler monkey deaths. In the same year, 715 suspected cases were investigated confirming YF in 0.7% of them. For these reasons, between 2002 and 2004, Venezuela's MOH has vaccinated approximately 1.9 million people in areas considered to be enzootic. The country's goal for 2006 is to have 7 million people residing in high-risk cities and towns vaccinated, and in this way, preventing and controlling this emerging zoonotic disease. PMID- 17135495 TI - Risk analysis and bovine tuberculosis, a re-emerging zoonosis. AB - The widespread of immunodeficiency with AIDS, the consequence of poverty on sanitary protection and information at both individual and state levels lead control of tuberculosis (TB) to be one of the priorities of World Health Organization programs. The impact of bovine tuberculosis (BTB) on humans is poorly documented. However, BTB remains a major problem for livestock in developing countries particularly in Africa and wildlife is responsible for the failure of TB eradication programs. In Africa, the consumption of raw milk and raw meat, and the development of bushmeat consumption as a cheap source of proteins, represent one of the principal routes for human contaminations with BTB. The exploration of these different pathways using tools as participatory epidemiology allows the risk analysis of the impact of BTB on human health in Africa. This analysis represents a management support and decision tool in the study and the control of zoonotic BTB. PMID- 17135496 TI - Status of mastitis as an emerging disease in improved and periurban dairy farms in India. AB - While reviewing present status of mastitis in India, results of investigations from periurban dairy farms on epidemiological, clinical manifestations, diagnosis, bacterial isolation, in vitro drug sensitivity, and treatment of subclinical mastitis have been presented. Mastitis, on account of its causing serious wastage and undesirable milk quality, is emerging as a major challenge among the others (like breeding improvement, nutrition management, control of infectious, tick-borne, blood, and internal parasitic diseases) in dairy development of tropics. Subclinical mastitis was found more important in India (varying from 10-50% in cows and 5-20% in buffaloes) than clinical mastitis (1 10%). The incidence was highest in Purebred Holsteins and Jerseys and lowest in local cattle and buffaloes. An investigation on 250 animals from periurban farms indicated that the monsoon season was more prone to subclinical mastitis than summer or winter, prevalence increased with higher lactation number and animals in 4th-5th month of lactation were found more susceptible (59.49%), hind quarters were found more affected (56.52%) than fore quarters (43.47%). The factors like herd size, agro climatic conditions of the region, variations in sociocultural practices, milk marketing, literacy level of the animal owner, system of feeding, and management were found important affecting the incidence of subclinical mastitis. Comparison of efficacy of different diagnostic techniques for subclinical mastitis, such as modified California mastitis test (MCMT), bromo thymol blue (BTB), modified whiteside test, trypsin inhibition test, milk pH, and electric conductivity indicated MCMT to be most sensitive (95.16%) and specific (98.02%) test. The antimicrobial susceptibility test revealed that most of the bacterial strains (gram positive, gram negative, and mixed) isolated from subclinical mastitis milk samples, were highly sensitive to enrofloxacin 53.91%, least sensitive to oxytetracycline 17.39% and ampicillin 7.83%, and resistant to streptomycin. The therapy with enrofloxacin and nimesulide was found more efficacious (92.30%) in treating subclinical mastitis cows. It was concluded that consideration of socioecological factors for mastitis control in periurban area would help to reduce the cost of mastitis control in clean milk production. PMID- 17135497 TI - Role of import and export regulatory animal health officials in international control and surveillance for animal diseases. AB - The challenges to those who regulate the import and export of animals and animal products are escalating, due to the evolving nature of animal and human disease agents. The diseases and agents of interest may include low pathogenic avian influenza, bluetongue, bovine spongiform encephalopathy, and foot-and-mouth disease. Fear of an incursion of an unknown or incompletely understood threat can significantly limit risk tolerance. The fear may be that an incursion will affect export trade or tourism. An incomplete knowledge of the animal health situation in the exporting country, due to insufficient surveillance for the disease agent of concern, may limit the application of science in import decisions. In addition, the disease agent may be inappropriately considered exotic if it has not been described. As a result, excessive safeguards for disease agents that do not present any new threat may be employed. To confront these challenges, we are striving toward transparency in international reporting. Moreover, regulatory import decisions exceeding the recommendations of the Terrestrial Animal Health Code and the Aquatic Animal Health Code of the World Organization for Animal Health must be fair and science-based. PMID- 17135498 TI - Regional and international approaches on prevention and control of animal transboundary and emerging diseases. AB - Transboundary animal diseases pose a serious risk to the world animal agriculture and food security and jeopardize international trade. The world has been facing devastating economic losses from major outbreaks of transboundary animal diseases (TADs) such as foot-and-mouth disease, classical swine fever, rinderpest, peste des petits ruminants (PPR), and Rift Valley fever. Lately the highly pathogenic avian influenza (HPAI) due to H5N1 virus, has become an international crisis as all regions around the world can be considered at risk. In the past decades, public health authorities within industrialized countries have been faced with an increasing number of food safety issues. The situation is equally serious in developing countries. The globalization of food (and feed) trade, facilitated by the liberalization of world trade, while offering many benefits and opportunities, also represents new risks. The GF-TADs Global Secretariat has carried out several regional consultations for the identification of priority diseases and best ways for their administration, prevention and control. In the questionnaires carried out and through the consultative process, it was noted that globally, FMD was ranked as the first and foremost priority. Rift Valley fever, and today highly pathogenic avian influenza, are defined as major animal diseases which also affect human health. PPR and CBPP, a disease which is particularly serious in Africa and finally, African swine fever (ASF) and classical swine fever (CSF) are also regionally recognised as top priorities on which the Framework is determined to work. The FAO philosophy--shared by the OIE- embraces the need to prevent and control TADs and emerging diseases at their source, which is most of the time in developing countries. Regional and international approaches have to be followed, and the FAO and OIE GF-TADs initiative provides the appropriate concepts and objectives as well as an organizational framework to link international and regional organizations at the service of their countries to better prevent and control the risks on animal and human health and the economic impact of TADs and emerging animal diseases. PMID- 17135499 TI - Linking human and animal health surveillance for emerging diseases in the United States: achievements and challenges. PMID- 17135500 TI - Development of a monoclonal antibody-based immunohistochemistry method for BSE surveillance in China. AB - Five hybridoma cell lines secreting anti-PrP antibodies were established from the fusion between mouse myeloma Sp2/0 and spleen cells from mice immunized with recombinant Chinese Luxi yellow cattle (Bos taurus, Luxi) PrP (24-234) or recombinant Chinese small-tailed Han sheep PrP (94-227). According to their Western blot reactivity, five monoclonal antibodies (mAbs) could be divided into two groups. Group A, mAbs 1H2, 4C6, and 4C11 recognized re-PrP, PrP(C), and PrP(Sc) from both bovine and sheep. Group B, mAbs 2H3 and 4H10 only recognized re PrP and PrP(Sc) of sheep, and especially, these two mAbs could not recognize PrP(C) from both bovine and sheep. In immunohistochemistry (IHC) test, mAb 4C11 immunostained the PrP(Sc) accumulation in tissue sections from BSE cattle and Scrapie sheep, and compared with mAb 6H4, it had the same immunohistochemical pattern. An IHC method based on mAb 4C11 for the detection of BSE was established and had been applied for the long-term surveillance of BSE in China. From 2001 to 2004, 12,692 samples from the whole country had been tested and all had negative results. PMID- 17135501 TI - A serological survey of pigs, horses, and ducks in Nepal for evidence of infection with Japanese encephalitis virus. AB - Japanese encephalitis (JE) is an emerging disease of animals and humans in Nepal. A serological study for antibody to JE virus was conducted in Nepal from September 2003 to August 2004 by collecting 280 sera from pigs, ducks, and horses covering 10 districts of the country. These sera were tested by performing competitive enzyme-linked immunosorbent assay for the detection of antibodies against JE virus. The total number of tested sera was 280, of which 43.92% were found positive for the presence of antibodies against JE virus infection in Nepal. Sero-prevalence of JE in pigs, ducks, and horses was 48.11%, 26.79%, and 50.0%, respectively. PMID- 17135502 TI - Porcine innate and adaptative immune responses to influenza and coronavirus infections. AB - Both innate and adaptative immune responses contribute to the control of infectious diseases, including by limiting the spreading of zoonotic diseases from animal reservoirs to humans. Pigs represent an important animal reservoir for influenza virus infection of human populations and are also naturally infected by coronaviruses, an important group of viruses, which includes the recently emerged severe acute respiratory syndrome (SARS) virus. Studies on both innate and adaptative immune responses of pigs to influenza virus and coronaviruses contribute, therefore, to a better control of these infections in their natural hosts and will be briefly reviewed in this article. Pro inflammatory cytokines, including type I interferon (IFN), tumor necrosis factor alpha (TNF-alpha), and interleukin-6 (IL-6), were found in lung secretions of influenza virus infected pigs, and correlated with the intensity of clinical signs, whereas prior vaccination against influenza strongly reduced the production of infectious virus and cytokines in the lungs upon challenge, which was associated with clinical protection. An early type I IFN production was also found in coronavirus infected pigs, including at mucosal sites. IFN induction by coronavirus is shown to involve interaction between a viral glycoprotein and a leukocyte subset, likely equivalent to plasmacytoid dendritic cells, present in the mucosae and associated lymphoid tissues. Given the IFN mediated antiviral and immunomodulatory effects, the use of IFN or IFN inducers may prove an efficient strategy for a better control of influenza virus and coronavirus infections in pigs. Because influenza and coronaviruses target mucosal surfaces, adaptative immune responses have to be characterized at mucosal sites. Thus, nasal and pulmonary antibody responses were analyzed in influenza virus infected or vaccinated pigs showing short-lived, but potentially protective local IgA and IgG antibody (Ab) responses. Interestingly, primary influenza virus infection induced long-lived increase of lung CD8(+) T cells and local lymphoproliferative responses. Pigs infected by a respiratory coronavirus (PRCV) showed virus specific IgG Ab-secreting cells in the bronchial lymph nodes, whereas the transmissible gastroenteritis coronavirus (TGEV) induced more IgA Ab-secreting cells in gut tissues, which illustrates the importance of the route of antigen administration for inducing local immune effector mechanisms. Porcine viral infections provide, therefore, valuable models for evaluating the immune parameters that are important for controlling transmission of important viral zoonotic infections. PMID- 17135503 TI - Ecopathological approach in tropical countries: a challenge in intensified production systems. AB - From the 1960s, in developed countries, epidemiological approach widened in veterinary sciences in order to understand and analyze the emergence of "production diseases" in "modern farms" where animal management was compared to industrial process. This approach was linked to "productivism" in livestock farming system. In France, this approach included formerly the understanding of farmers' practices and considered the health disorders as an output of farming system. This theory was known as "ecopathological approach." Since 2000, the main purpose for veterinary sciences seems to be now emerging diseases. In tropical countries, a high intensification of livestock production is expected, especially in periurban areas. This intensification linked to a general better control of main infectious diseases (i.e., Rinderpest, CBPP, anthrax) in such farming system leads to many changes in the farming practices and is associated to the emergence of production diseases as mastitis or infertility in dairy farms, respiratory diseases in pig farms and so on. In those conditions, it is suggested to initiate ecopathological approach in these intensified systems in order to identify the main risk factors due to farmers' practices, environmental conditions, and herd characteristics. The general methodology and some examples in tropical countries (Chad, Brazil, and Senegal) are proposed in the present article. PMID- 17135504 TI - Integrated risk reduction along the food chain. AB - Animal health is the crucial first part of the food chain and must be considered when developing the controls or preventative measures for an endemic or emerging zoonotic food-borne diseases. Increasing the number of complementary control measures at various points along the food processing chain results in overall risk reduction and improved safety of products for the domestic markets and trade. In addition, a risk assessment must be made and surveillance implemented. Measures require sufficient infrastructure within veterinary services, and must be controlled and re-evaluated periodically. The system must be communicated to stakeholders in order to improve compliance as well as confidence from domestic consumers, trading partners, and the international community. Bovine spongiform encephalopathy (BSE) and avian influenza (AI) can be used as examples for implementation of these control concepts. PMID- 17135505 TI - History and evolution of HPAI viruses in southeast Asia. AB - Highly pathogenic avian influenza (HPAI) has been recognized as a serious viral disease of poultry since 1878. The number of outbreaks of this disease globally has increased in the past 10 years culminating in 2004 with the unprecedented outbreak of H5N1 HPAI involving nine countries in East and South East Asia. Apart from the geographical extent of this outbreak and apparent rapid spread, this epidemic has a number of unique features, among which is the carriage of highly pathogenic AI viruses by asymptomatic domestic waterfowl. When this disease first emerged it was recognized almost simultaneously in a number of countries for the first time. This created considerable concern among both veterinary and public health authorities especially as the virus was also shown to cause fatal disease in humans. This article brings together a range of information on H5N1 HPAI viruses in Asia that were collected by FAO during the past year through field projects and explores possible reasons for the emergence of the disease in late 2003 and early 2004. Key epidemiological features of the disease in different Asian countries are described in an attempt to look for, and where possible, explain similarities and differences. This includes assessment of factors that could have contributed to the spread of the disease. Molecular aspects of the viruses are examined to assess relationships between isolates from different locations and times so as to gain insights into the origins of viruses in various countries. It is apparent that the coincidence and grouping of the reports declaring the outbreaks of HPAI did not truly reflect the time course of disease emergence, which was widespread well before the outbreak. The factors that could have led to a change from infection to emergence of widespread disease in 2003 2004 are discussed. There are still some questions that remain unanswered regarding the origins of the 2004 outbreak. This article does not provide answers to all of these, but brings together what is currently known about these outbreaks and the viruses that have caused them. PMID- 17135506 TI - Surveillance for avian influenza in the United States. AB - In the United States, some 1.7 million agar gel immuno diffusion (AGID) tests for avian influenza (AI) are conducted yearly by various poultry groups, governmental sectors, and private industry. In addition to the AGID test, additional testing includes virus isolations, enzyme-linked immunosorbent assays, real-time reverse transcriptase polymerase chain reactions, and hemagglutination inhibition (HI) tests. HI and neuraminidase inhibition tests are conducted on positive AGID samples to determine the subtype. Directigen, a type of antigen capture test, is used in the field in some cases. If monitoring and surveillance activities give rise to a suspicious test result, the accredited veterinarian and official State laboratory are required to report these to the governmental authorities. A thorough investigation in collaboration with the National Veterinary Services Laboratories (a World Organization for Animal Health--AI reference laboratory), State and Federal veterinarians, and others is conducted. Testing conducted as part of the National Poultry Improvement Plan (NPIP) effectively monitors the status of breeder and multiplier flocks. A new commercial poultry program is being added and will expand NPIP AI testing to all commercial flocks. Private poultry companies conduct additional tests; and in the poultry-producing States, there are active state-wide programs to monitor poultry health. All components of the live-bird market system (source flocks, haulers, dealers, and markets) are tested under the Low Pathogenicity AI Live-Bird Market Program. PMID- 17135507 TI - Influenza a virus surveillance of migratory waterfowl in Barbados, West Indies. PMID- 17135508 TI - Isolation of avian influenza virus A subtype H5N1 from internal contents (albumen and allantoic fluid) of Japanese quail (Coturnix coturnix japonica) eggs and oviduct during a natural outbreak. AB - Avian influenza virus (AIV) was recovered from the internal contents of eggs, including mixture of albumen and allantoic fluid, and from the oviduct of naturally infected Japanese quail (Coturnix coturnix japonica) flocks in the southern part of Thailand. The virus titers of 10(4.6)-10(6.2) ELD(50)/mL were directly measured from the internal content of infected eggs. The virus was isolated by chorioallantoic sac inoculation of embryonating chicken eggs. Infected allantoic fluid was identified as hemagglutinating virus and then was indicated the presence of H5 hemagglutinin. The virus was confirmed to be H5N1 subtype influenza A virus by reverse transcriptase-polymerase chain reaction. Additionally, real-time reverse transcriptase-polymerase chain reaction assay could specifically detect influenza virus subtype H5. Furthermore, indirect fluorescent antibody (IFA) test by using specific anti-influenza A monoclonal antibody indicated that virus antigens were detected in the parenchyma of multiple tissues. Systemic localization of viral antigen detected was certainly considered to be viremic stage. In addition, influenza virus antigen was also detected by IFA in allantoic fluid sediments isolated from internal content of egg or oviduct. The conclusion of isolated AIV type A subtype H5N1 from these two infected materials was correlated to the viremic stage of infection because the virus antigens could be observed in almost all tissues. Conclusively, the need for adequate safeguards to prevent contamination and spread of the virus to the environment during movement of eggs--including hatching eggs, cracked eggs, and other relevant infected materials-- or egg consumption from area of outbreak is emphasized and must not be ignored for the reasons of animal, public, and environmental health. PMID- 17135509 TI - Principles for vaccine protection in chickens and domestic waterfowl against avian influenza: emphasis on Asian H5N1 high pathogenicity avian influenza. AB - The H5N1 highly pathogenic (HP) avian influenza (AI) epizootic began with reports of mortality from China in 1996 and, by June 2005, caused outbreaks of disease in nine additional Asian countries, affecting or resulting in culling of over 200 million birds. Vaccines can be used in programs to prevent, manage, or eradicate AI. However, vaccines should only be used as part of a comprehensive control strategy that also includes biosecurity, quarantine, surveillance and diagnostics, education, and elimination of infected poultry. Potent AI vaccines, when properly used, can prevent disease and death, increase resistance to infection, reduce field virus replication and shedding, and reduce virus transmission, but do not provide "sterilizing immunity" in the field; i.e., vaccination does not completely prevent AI virus replication. Inactivated AI vaccines and a recombinant fowlpox-H5-AI vaccine are licensed and used in various countries. Vaccines have been shown to protect chickens, geese, and ducks from H5 HPAI. The inactivated vaccines prevented disease and mortality in chickens and geese, and reduced the ability of the field virus to replicate in gastrointestinal and respiratory tracts. Although the Asian H5N1 HPAI virus did not cause disease or mortality in ducks, the use of inactivated vaccine did reduce field virus replication in the respiratory and intestinal tracts. The inactivated vaccine protected geese from morbidity and mortality, and reduced challenge virus replication. The recombinant fowlpox-H5-AI vaccine has provided similar protection, but the vaccine is used only in chickens and with the advantage of application at 1 day of age in the hatchery. PMID- 17135510 TI - Vaccines developed for H5 highly pathogenic avian influenza in China. AB - Since the first detection of highly pathogenic H5N1 avian influenza virus from sick goose in Guangdong province in China in 1996, scientists in China started to develop vaccines for avian influenza pandemic preparedness. An H5N2 inactivated vaccine was produced from a low pathogenic virus, A/turkey/England/N-28/73, and was used for the buffer zone vaccination in the H5N1 outbreaks in 2004 in China. We also generated a low pathogenic H5N1 reassortant virus A/Harbin/Re-1/2003 (Re 1) that derives its HA and NA genes from GSGD/96 virus and six internal genes from the high-growth A/Puerto Rico/8/34 (PR8) virus by using plasmid-based reverse genetics. The inactivated vaccine derived from Re-1 strain could induce more than 10 months protective immune response in chickens after one dose inoculation, and most importantly, this vaccine is immunogenic for geese and ducks. An H5N1 fowlpox vectored live vaccine was also generated by inserting the HA and NA genes of GSGD/96 virus in the genome of a fowlpox vaccine strain. Laboratory tests indicated that after one dose of immunization of this vaccine, chickens could develop an over than 40 weeks protective immune response against H5N1 virus challenge. PMID- 17135511 TI - Development and use of fowlpox vectored vaccines for avian influenza. AB - The avian influenza (AI) vaccine designated TROVAC-AIV H5 (TROVAC-H5) contains a live recombinant fowlpox rec. (FP) recombinant (recFP), expressing the hemagglutinin (HA) gene of an AI H5 subtype isolate. This recombinant vaccine was granted a license in the United States for emergency use in 1998 and full registration in Mexico, Guatemala, and El Salvador where over 2 billion doses have been administered. One injection of TROVAC-H5 protects chickens against AI induced mortality and morbidity for at least 20 weeks, and significantly decreases shedding after challenge with a wide panel of H5-subtype AI strains, regardless of neuraminidase subtype. Recently, excellent protection was demonstrated against 2003 and 2004 Asian highly pathogenic H5N1 isolates. Whereas TROVAC-H5 AI H5 efficacy was not inhibited by anti-AI or anti-fowlpox maternal antibodies (passive immunity), protection to AI was significantly decreased in chickens previously vaccinated or infected with FP (active immunity). Advantages of the TROVAC-H5 vaccine over inactivated AI vaccines are: (a) single administration at 1 day of age and early onset (1 week) of protection, (b) easy monitoring of AI infection in vaccinated flocks with agar gel precipitation (AGP) and enzyme-linked immunosorbent assay (ELISA) used as tests to differentiate infected from vaccinated animals (DIVA tests), and (c) no residue problem due to adjuvant. These features make TROVAC-H5 an ideal AI vaccine for routine administration of day-of-age chicks in hatcheries. RecFP expressing HA from three lineages of H7 subtype (Eurasian, American, and Australian) were also tested for efficacy against a highly pathogenic avian influenza (HPAI) Eurasian HPAI H7N1. Only the recFP expressing the Eurasian H7 gene provided sufficient protection indicating that the breadth of protection induced by recFP is apparently restricted for H7 isolates. The fowlpox vector technology can also be used for the production of an emergency vaccine: once the HA sequence of an emerging AI virus is known, recFP can be rapidly generated. TROVAC-H5 has recently been shown to be immunogenic in cats and could therefore also be considered for use in mammals. PMID- 17135512 TI - SEPPIC vaccine adjuvants for poultry. AB - Two inactivated antigens (Newcastle and Pasteurella Multocida) were formulated with different adjuvants and tested in two separate experiments in poultry. Oil formulations constituting water in oil (W/O) or water in oil in water (W/O/W) emulsions were assessed for antibody response, protection, local reactions, and vaccine physicochemical parameters. Robust, efficacious, and safe formulations were obtained with W/O formulations whereas W/O/W was especially safe with maintained efficacy. Results show that it is possible to improve traditional Tween Span formulations for safety and efficacy parameters by using Montanide ISA 70 for W/O formulations and Montanide ISA 206 for W/O/W when safety is the priority. PMID- 17135513 TI - West Nile virus in Guadeloupe: introduction, spread, and decrease in circulation level: 2002-2005. AB - In July 2002, a surveillance system was implemented on Guadeloupe to detect for the potential introduction and monitor the spread of West Nile virus (WNV). From 2002 to 2004, equines and chickens were serologically assayed for antibodies to WNV by IgG and IgM enzyme-linked immunosorbent assay (ELISA), epitope-blocking ELISA, and plaque reduction neutralization tests. After introduction, probably through migratory birds at the end of 2001, many seroconversions occurred between July and October 2002 resulting in a high seroprevalence (19.3%) in equines in 2003. WNV circulation levels decreased dramatically in 2003 and 2004 as assessed by the absence of seroconversion in equine and the very low prevalence in chickens. This decrease coincided with a 7-month drought that presumably caused a decrease in vector populations. In 2005, a sentinel survey was implemented in equines and chickens placed in areas at high risk and the very low rate of seroconversion (1 equine out of 106, no chicken) demonstrated that WNV circulation is now occurring at a very low level. PMID- 17135514 TI - Serological assessment of West Nile fever virus activity in the pastoral system of Ferlo, Senegal. AB - The Ferlo area (north-central Senegal) is characterized by a system of temporary ponds favorable to arboviruses among which West Nile fever (WNF) was already identified. During the rainy season in 2003, a serological study was undertaken on horses to assess the activity of the WNF virus (WNFV) in Barkedji (Ferlo). The observed serological prevalence rate was 78.3% for neutralizing antibodies, with a 95% confidence interval (CI) of [64.0, 92.7]. This prevalence rate significantly increased with age (P = 10(-5)). This study confirmed that WNF was endemic in the Ferlo. The transmission risks depended on the introduction of the WNFV in the ecosystem--probably with migrating birds, on its amplification in hosts and on the vector-population dynamic. Further studies are needed to investigate how the cycle is initiated in Barkedji at the beginning of the rainy season and the impact of climatic variations on the risk of transmission of WNF. A surveillance system should be implemented: (a) to assess the clinical impact of the WNF on human and equine populations, (b) to provide an early detection of virulent strains, and (c) to assess the risk of WNF transmission to disease-free ecosystems via migrating birds. PMID- 17135515 TI - Recommendations from the avian influenza vaccine workshop. PMID- 17135516 TI - Genotyping of Newcastle disease viruses isolated from 2002 to 2004 in China. AB - The main function region of the fusion (F) protein gene of 124 strains of Newcastle disease virus isolated from 2002 to 2004 in China was amplified and sequenced for further phylogenetic and residue substitutive analysis. Most of the isolates were classified into genotype VIIc, VIId, VIf, and VIb, while others into genotype IX, III, or II. The genotype IX, a unique genotype which includes strain F48, the first Chinese virulent NDV strain isolated in 1948, were still found inducing sporadic infections in certain areas. Subgenotype VIIc, VIId, and VIIe viruses, which were distributed in clusters in the phylogenetic tree distinct from members of subgenotypes VIIa and VIIb, were responsible for most outbreaks in China and circulated predominantly in China in recent years. Strain NDV03-026, an isolate of the genotype II which was normally lentogenic, was found carrying (112)RRQKRF(117) motif at the cleavage site of F protein as the virulent strain. PMID- 17135517 TI - Descriptive and spatial epidemiology of Rift valley fever outbreak in Yemen 2000 2001. AB - Rift valley fever (RVF) is an arboviral disease produced by a bunyavirus belonging to the genus Phlebovirus. Several species of Aedes and Culex are the vectors of this virus that affects sheep, goats, buffalos, cattle, camels and human beings. The human disease is well known, especially during periods of intense epizootic activity. The initial description of the disease dates back to 1930, when animals and human outbreaks appeared on a farm in Lake Naivasha, in the Great Rift Valley of Kenya. Until 2000, this disease was only described in Africa, and then outbreaks were also declared in the Kingdom of Saudi Arabia (2000-2001 and 2004) and in Yemen (2000-2001). Animal and human cases were recorded. This work presents a retrospective summary of the data collected on animal RVF cases during this epidemic in Yemen. Results from several RVF surveys were gathered from the Yemeni vet services and FAO experts. Geographical data (topographic maps and data freely available on internet) were used for the location of outbreaks. After cleaning and standardization of location names, all the data were introduced into a GIS database. The spatial distribution of outbreaks was then studied at two scales: at the national level and at a local scale in the particular area of Wadi Mawr in the Tihama plain, Western coast of Yemen. PMID- 17135518 TI - Efficacy of DNA immunization with F and G protein genes of Nipah virus. AB - We investigated the antibody response of DNA immunization with two mammalian codon optimized envelope glycoprotein genes, F and G, of Nipah virus in a mouse model. The results indicated that G gene immunization elicited more significant specific serum IgG response and neutralization antibody response than F gene did, suggesting that the G gene DNA immunization is a potential vaccine strategy against Nipah virus. PMID- 17135519 TI - Generating vesicular stomatitis virus pseudotype bearing the severe acute respiratory syndrome coronavirus spike envelope glycoprotein for rapid and safe neutralization test or cell-entry assay. AB - We generated a recombinant vesicular stomatitis virus (VSV) pseudotype (VSV Delta G*SG) by replacing the envelope G gene with the GFP gene and complementing with spike glycoprotein (S) of SARS-CoV in trans. The neutralization and infection blocking tests showed that the VSV Delta G*SG and SARS-CoV reacted similarly to SARS-CoV specific antiserum, suggesting the VSVDelta G*SG can be a safe replacement of the live SARS-CoV for neutralization test and cell-entry assay. PMID- 17135520 TI - Prevalence of Salmonella species in various raw meat samples of a local market in Kathmandu. AB - A cross-sectional study of raw meat samples from the local meat market of Kathmandu Metropolitan City was carried out during September 2002 to May 2003 with special emphasis on isolation and identification of Salmonella bacteria. A total of 123 raw meat samples (55 chicken, 37 buffalo, and 31 goat) were collected and analyzed relative to season. Salmonella spp was found in 11.4% (14/123) meat samples. Eight samples of chicken, that is, 14.5%, five samples of buffalo (13.5%), and one sample of goat (3.3%) were found to be positive for Salmonella. Salmonella prevalence revealed Salmonella (S.) pullorum in 3.3% samples, S. gallinarum in 0.8%, S. typhi in 1.6%, S. choleraesuis in 0.8%, and Salmonella of subgenus I or II group in 4.9% samples. More than 80% meat samples microbiologically processed indicated coliform contamination. Seasonal prevalence of Salmonella was highest in the months of April/May. Surveys revealed unsatisfactory conditions of sanitation in the local meat markets of Kathmandu. PMID- 17135521 TI - Prevalence of Salmonella in retail chicken meat in Hanoi, Vietnam. AB - Infection with Salmonella (S.) is the most frequently reported cause of bacterial food-borne illness worldwide. Poultry are a common source and, in recent years, much attention has been focused in determining the prevalence of Salmonella during the different stages in the poultry production chain. This article was designed to investigate the prevalence of Salmonella serovars in retail chicken meat sold in Hanoi. A total of 262 samples were randomly collected from retail markets and examined for the presence of Salmonella. Of these samples, 48.9% were found to be contaminated with Salmonella. Predominant serotypes were S. Agona, S. Emek, S. London. The prevalence of S. Enteritidis and S. Typhimurium was considered. These findings have highlighted the magnitude of Salmonella contamination in retail chicken meat in Hanoi. On the basis of these preliminary survey results, it is recommended that a cost-effective monitoring and surveillance system for Salmonella should be established in Hanoi. This system should be augmented by good agricultural and hygienic practices and well-designed longitudinal research activities on the whole poultry production chain. PMID- 17135522 TI - Study of Salmonella, Campylobacter, and Escherichia coli contamination in raw food available in factories, schools, and hospital canteens in Hanoi, Vietnam. AB - This study on the contamination rates of raw foods available in factory, school, and hospital canteens in Hanoi, Vietnam, with the bacteria of Salmonella, Campylobacter, and Escherichia coli (E. coli) was carried out between 2003 and 2004. A total of 177 raw food samples of vegetables, meat (beef and pork), fish, and poultry were examined to provide baseline data for evaluation of microbiological risks in general, and identification of potential vehicles for pathogenic cross-contamination in canteens. The study confirmed that unprocessed fish and poultry are likely to be contaminated with Salmonella and in the absence of proper kitchen hygiene and may contaminate processed foods. Raw poultry samples were highly contaminated with E. coli (45%), Campylobacter jejuni (C. jejuni) (28.3%), and Salmonella (8.3%) and classified as high-risk food. E. coli was also detected in raw meat, fish, and vegetables with the rate of 21.3%, 6.6%, and 18.5%, respectively. This article confirmed the importance of hygienic working practices when preparing food. PMID- 17135523 TI - Prevalence of Salmonella spp. in poultry in Vietnam. AB - The prevalence of Salmonella spp. in chickens and ducks from North, Central, and South Vietnam was followed over a 4 year period. Several different analyses were employed and the current prevalence was shown to be less than in previous studies. PMID- 17135524 TI - Prevalence and epidemiology of Salmonella spp. in small pig abattoirs of Hanoi, Vietnam. AB - The prevalence of Salmonella spp. in pigs was evaluated in a survey of small abattoirs in Hanoi, Vietnam. Cecal contents, carcass swabs, and tank water samples were collected for bacterial isolation in various media. Prevalence rates exceeded 50% in pig samples and 62% in water samples. This increased prevalence indicates the need for risk assessment evaluations along the entire production chain. PMID- 17135525 TI - Study on the prevalence of Campylobacter spp. from chicken meat in Hanoi, Vietnam. AB - Campylobacter spp. is considered to be the most common bacterial cause of human gastroenteritis worldwide. In developing countries, Campylobacter spp. diarrhea is an important cause of childhood morbidity. Chicken meat is known to be a major source of Campylobacteriosis infection in the world. The purpose of this study was to investigate the prevalence of Campylobacter spp. in chicken meat. A total of 100 samples from breast part of chicken carcass were collected from retail market in Hanoi. The samples were taken for bacteriological analysis following the ISO 10272 standards. Thirty one samples (31%) were found positive for Campylobacter spp. The most frequently isolated Campylobacter was Campylobacter jejuni (45.2%) followed by Campylobacter coli (25.8%). Due to high contamination rates of retail chicken products, special attention must be paid to good manufacturing practices of food processors and vendors. Further studies should be done to assess the risk factors of Campylobacter spp. contamination in the Vietnamese fowl production chain. PMID- 17135526 TI - Characterization of lactic acid bacteria and other gut bacteria in pigs by a macroarraying method. AB - Lactic acid bacteria (LAB) consist of many genera, Gram-positive, and nonspore forming micro-organisms; some members being used as probiotics while some others have negative effects on pig health. Bacterial species in the gastrointestinal tract can produce antibacterial substances, reduce serum cholesterol in their host, or can be responsible for growth reduction, diarrhea, and intestinal epithelial damage. It is therefore important for the pig industry to evaluate the impact of food and farm management on the presence of "good" or "bad" bacteria and the risk for consumers. This articles focuses on the molecular identification of gut microflora species following different diets given to pigs in UK and correlating the data on growth, health, and welfare. First of all, pig feces were individually collected from sows before and after farrowing and also from piglets before and after weaning over several months. Bacteria colonies were grown on MRS agar plates from feces and DNA was extracted (QIAamp DNA stool kit) and amplified using 16S rDNA (27f and 519r) primers. DNA sequencing and sequence alignment allowed us to identify species-specific zones, which were used as probes in a macroarray system also known as reverse line blot hybridization. Some probes were found to be species specific for the following species: Lactobacillus acidophilus, L. animalis, L. gallinarum, L. kitasanotis, L salivarius, Streptococcus alactolyticus, S. hyointestinalis, and Sarcina ventriculi. Actual studies are now focusing on the impact of diets of the microflora in different gut parts and at different stages of the animal's life. PMID- 17135527 TI - Porcine eperythrozoonosis in China. AB - Eperythrozoonosis of swine (also designated as porcine mycoplasmosis) is a disease of swine under stress, expressed as a febrile condition with development of an acute ictero-anemia. It is caused by Eperythrozoon suis and usually causes a subclinical infection with a latent carrier state that persists for extended periods. In China, this disease has gradually developed as an important intercurrent disease and an emerging swine disease that, in recent years, has spread throughout all provinces except Tibet. Classical swine fever (hog cholera), porcine influenza, swine enzootic pneumonia, porcine reproductive and respiratory syndrome (blue ear disease), streptococci, and toxoplasmosis were detected in Eperythrozoonosis-infected pig herds, and caused serious economic losses. National epidemiology surveillance in 2002 revealed that this disease caused a total morbidity of 30% and a mortality of 10-20%. Total mortality (which includes culling sick pigs) was more than 60%. The morbidity within infected herds was near 100%, has spread throughout with a total mortality rate usually over 50%. Mortality of piglets in some districts was as high as 50%. The highest infection rate on pig farms was more than 90%. The farms with higher infection rates occurred in pig-raising areas during epidemic seasons. New diagnostic tests, such as ELISA and PCR, have been developed for the detection of porcine eperythrozoonosis, but traditionally the diagnosis of the disease is still based on clinical history and optical microscopic examination of the causative agent in blood smears. Efficient preventive and control measures include the detection of carriers in pig herds and treatment of sick pigs with drugs, such as long-acting oxytetracycline, doxycycline, or aceturate of diminazene. Oxytetracyclines as feed additives have been introduced for eperythrozoonosis prevention in uninfected pig herds, and pig producers have taken measures to reduce stress and improve sanitary conditions. PMID- 17135528 TI - Bovine transcriptome analysis by SAGE technology during an experimental Trypanosoma congolense infection. AB - In central and sub-Saharan Africa, trypanosomosis is a tsetse fly-transmitted disease, which is considered as the most important impediment to livestock production in the region. However, several indigenous West African taurine breeds (Bos taurus) present remarkable tolerance to the infection. This genetic capability, named trypanotolerance, results from numerous biological mechanisms most probably under multigenic dependences, among which are control of the trypanosome infection by limitation of parasitemia and control of severe anemia due to the pathogenic effects. Today, some postgenomic biotechnologies, such as transcriptome analyses, allow characterization of the full expressed genes involved in the majority of animal diseases under genetic control. One of them is serial analysis of gene expression (SAGE) technology, which consists of the construction of mRNA transcript libraries for qualitative and quantitative analysis of the entire genes expressed or inactivated at a particular step of cellular activation. We developed four different mRNA transcript libraries from white blood cells on a N'Dama trypanotolerant animal during an experimental Trypanosoma congolense (T. congolense) infection: one before experimental infection (ND0), one at the parasitemia peak (NDm), one at the minimal packed cell volume (NDa), and the last one at the end of the experiment after normalization (NDf). Bioinformatic comparisons in bovine genomic databases allowed us to obtain more than 75,000 sequences, among which are several known genes, some others are already described as expressed sequence tags (ESTs), and the last are completely new, but probably functional in trypanotolerance. The knowledge of all identified named or unnamed genes involved in trypanotolerance characteristics will allow us to use them in a field marker-assisted selections strategy and in microarrays prediction sets for bovine trypanotolerance. PMID- 17135529 TI - Trypanosomosis in goats: current status. AB - Trypanosomosis is a major constraint on ruminant livestock production in Africa, Asia, and South America. The principal host species affected varies geographically, but buffalo, cattle, camels, and horses are particularly sensitive. Natural infections with Trypanosoma congolense, T. vivax, T. brucei, and T. evansi have been described in goats. Trypanosomosis in goats produces acute, subacute, chronic, or subclinical forms, being T. vivax, T. congolense, and T. evansi, the most invasive trypanosomes for goats. However, the role of goats in the epidemiology of trypanosomosis is largely discussed and not well understood. Thus, it has commonly been assumed that trypanosomosis presents a subclinical course and that goats do not play an important role in the epidemiology of the disease. This can partially be due to parasitemia caused by trypanosomes which has been considered low in goats. However, this assumption is currently undergoing a critical reappraisal because of goats may also serve as a reservoir of trypanosome infection for other species, including the human beings in the case of T. brucei rhodesiense. The present article describes the current status of trypanosomosis in goats in Africa, Asia, and South America. Pathogenesis, clinical features, diagnosis, and treatment of the different trypanosomes are also described. The possible role in the epidemiology of the disease in the different areas is also discussed. PMID- 17135530 TI - Detection of T.b. rhodesiense trypanosomes in humans and domestic animals in south east Uganda by amplification of serum resistance-associated gene. AB - The human serum resistance-associated (SRA) gene was identified in 28 (80%) of the 35 T.b. rhodesiense trypanosomes from parasitologically confirmed sleeping sickness cases, using the primers designed by Radwanska and in 27 (77.1%) of the same 35 T.b. rhodesiense trypanosomes using the primers designed by Gibson. However, about 20% of the 35 T.b. rhodesiense trypanosomes could not be detected by SRA-polymerase chain reaction (PCR) even when an aliquot of the first PCR was used in the second PCR, indicating that the gene may be absent in those trypanosomes or the trypanosomes could be having another variant of SRA not detectable by these primers since three variants of SRA genes have so far been identified or the amount of trypanosomal DNA extracted from infected blood was too low to be detected. The trypanosome isolates that are SRA gene negative may indicate the presence of some T.b. rhodesiense trypanosomes with modified or lack SRA genes or simple loss of the SRA gene from the expression site in which it resides during antigenic variation. Analysis of trypanosomes derived from domestic animals showed that 79 (90.8%) of the 87 trypanosomes isolated from cattle were positive by Trypanosoma brucei (TBR)-PCR, indicating that they were Trypanozoon while 8 (9.2%) of the trypanosome isolates which were negative by TBR PCR could be T. vivax, T. congolense, or T. theileri. When subjected to SRA-PCR, 10 (11.5%) of the 87 trypanosomes isolates derived from cattle were positive, indicating that there could be T.b. rhodesiense circulating in cattle, which is similar to the percentage of T.b. rhodesiense previously obtained in cattle in Serere, Soroti district. PMID- 17135531 TI - Trypanosomiasis in domestic animals of Makwanpur district, Nepal. AB - Trypanosomiasis is an infectious emerging hemoprotozoan parasitic disease in domestical animals of Nepal. The prevalence of disease was found in 16 of 240 (6.67%) in domestic animals of Makawanpur district, out of which 9 of 105 were (8.57%) cattle; 5 of 75 (6.67%) buffalos, and 2 of 15 (13.3%) dogs, while none of the goats and pigs acquired infection. The disease was found maximum during rainy season 9 of 82 (10.98%) with higher prevalence among cross breeds than that of local breeds. PMID- 17135532 TI - Prevalence of Trypanosoma evansi infection in equines and camels in the Punjab region, Pakistan. AB - A cross-sectional study has been carried out in order to determine the prevalence of Trypanosoma evansi infection in susceptible hosts in the Punjab region (Pakistan). A total of 170 equines and 150 dromedary camels were examined. Five (3.3%) and 6 (4%) camels were positive at parasitological and serological examination, respectively. None of the equines tested positive at any method. These results seem to indicate that T. evansi infection has a relatively low prevalence in the Punjab region. However, efforts must be done in order to establish control measures in affected herds and avoid dissemination of the disease. PMID- 17135533 TI - Clinical, hematological, and biochemical findings in an outbreak of abortion and neonatal mortality associated with Trypanosoma evansi infection in dromedary camels. AB - This article presents the clinical and laboratorial findings in an outbreak of abortions and high neonatal mortality attributable to Trypanosoma evansi infection in camels. A total of 16 females were diagnosed, 2 of which showed moderate signs of chronic form, particularly hyporexia and intolerance to exercise. The main laboratorial findings were regenerative anemia (hemolytic anemia), lymphocytic and monocytic leukocytosis, hyperproteinemia, hyperglobulinemia, hypoglycemia, serum urea increased, and serum iron decreased. The most characteristic finding in the examined females would be the uremia, probably due to the higher protein metabolism. PMID- 17135534 TI - Prevalence of Cryptosporidium among dairy cows in Thailand. AB - Cryptosporidium species are frequently associated with diarrhea among AIDS patients in Thailand, and dairy herds are a possible source of some of these infections. A cross-sectional study was performed to determine if Cryptosporidium is present among dairy cows in Thailand. Fecal samples were randomly collected from 363 Holstein-Friesian dairy cows from 108 of 860 farms in the Nong Pho region of central Thailand. The average prevalence of Cryptosporidium among dairy cows was 9.4%, according to an assay for Cryptosporidium-specific antigen (CSA) and 0.6% by microscopic examination of acid-fast stained feces. CSA was detected in all host age categories tested, but was most prevalent among calves (15.1%). Overall, 31.5% of farms were contaminated with Cryptosporidium infections. Fifty percent of poorly managed farms had CSA-positive cows, which were more likely to contaminate water and raw milk, while 12.9% of farms with acceptable management practices had CSA-positive cows. There was no association between the detection of Cryptosporidium and other gastrointestinal parasites. These results indicate that Cryptosporidium is enzootic among Thai dairy cattle, and suggest that cattle could have a role in zoonotic cryptosporidiosis in Thailand. PMID- 17135535 TI - Toxoplasmosis in piglets. AB - Seventeen-day-old piglets in a small holding farm in southern Thailand manifested signs of convulsion, fever, and death. The morbidity and mortality rate were approximately 26.09% (6/23) and 4.35% (1/23), respectively. Impression smear from lungs demonstrated tachyzoite stage of Toxoplasma gondii. Histopathological investigation revealed interstitial pneumonia. Further investigation, blood collection, educating the farmer, and prescribed affected herd with sulfa trimethoprim were performed soon after Toxoplasma infection was demonstrated. A serological detection of T. gondii infection among affected herds using latex agglutination test was conducted on 14 serum samples. The overall seroprevalence was 71.43% (10/14). The titers were 1:64 in 3 pigs (21.43%), 1:128 in 4 (28.57%), 1:256 in 2 (14.29%), and 1:512 in 1 (7.14%). A cat in this farm showed antibody titer 1:32 while a dog was seronegative. Data derived from this case indicated two possible routes of transmission: transplacental infection and ingested food or water contaminated with oocysts shed by cat in this farm. However, rodents can not exclude as a role of toxoplasmosis transmission. Serological monitoring at slaughter houses combine with good sanitary practices, rodent and cat control in the farm are important measures to prevent toxoplasma infection and improve human health in the future. PMID- 17135536 TI - Cysticercosis of slaughter cattle in southeastern Nigeria. AB - The incidence of cysticercosis due to Taenia saginata in both local and exotic breeds of cattle slaughtered for meat in southeastern Nigeria between November 1999 and April 2002 is reported. The examination of various organs of 25,800 cattle in 10 major abattoirs of this region showed that 6750 (26.2%) were infected with Cysticercus (C.) bovis. The prevalence rates varied from one abattoir to another while the rates of cysticercosis in local and exotic breeds varied significantly (P > 0.05). Sixty percent of all the infected animals had cysts. The tongue, cardiac, and masseter muscles were the main predilection sites of the cysts. Out of 11,720 male cattle, examined, 3215 (27.4%) had cysts of C. bovis while 160 (13.6%) of the 1180 female animals investigated were infected. There was an inverse relationship between the ages of the animals and prevalence of infection with C. bovis (r = -0.8743, P < 0.05). Monthly occurrence of the cysts in the animals revealed an upsurge of infected animals during the dry season. The epidemiology and epizootiology of Taenia saginata and C. bovis in relation to the veterinary service agencies and public health planners in southeastern Nigeria are highlighted. PMID- 17135537 TI - Use of Bacillus thuringiensis toxin as an alternative method of control against Haemonchus contortus. AB - The biocide activity of Bacillus thuringiensis (B. thuringiensis) IB-16 strain was evaluated against the blood-feeding nematode Haemonchus contortus (H. contortus) in vitro and in vivo assays. Twenty experimental jirds (Meriones unguiculatus) and 32 sheep were infected with H. contortus by oral route. Fourteen days post infection 10 jirds were treated with 0.2 mg per mL of IB-16 soluble toxin by intraperitoneal (i.p.) route and 8 sheep were treated with 0.5 mg of toxin per kg of body weight by intramuscular route 35 days after H. contortus infection. Same number of treated jirds and sheep were used as control. Fecal and blood samples were analyzed from experimental sheep to estimate the number of parasitic eggs, percentage of eosinophils, packed cell volume (PCV), and IgG title. All experimental jirds and 16 sheep (8 treated and 8 controls) were sacrificed at days 5, 7, and 30 after B. thuringiensis treatment. The percentage of nematode reduction was estimated from L(4) and H. contortus adults. The percentage of protection was 80.0 to jirds and 73.8 and 53.3 to sheep, sacrificed at days 7 and 30, respectively. Moreover, nonsignificant difference (P > or = 0.05) was detected from parasitic eggs, eosinophils, and PCV parameters. Significance level of IgG was observed only before H. contortus treatment with B. thuringiensis soluble toxin (P < or = 0.05) but nonsignificant difference was observed after treatment. These results showed that B. thuringiensis activity was similar to those observed by anthelmintic treatment and it could be used as an alternative biological method. PMID- 17135538 TI - Biological control of gastrointestinal parasitic nematodes using Duddingtonia flagrans in sheep under natural conditions in Mexico. AB - This investigation was aimed to evaluate the use of an oral bio-preparation containing Duddingtonia flagrans chlamydospores for the control of sheep gastrointestinal parasitic nematodes under the Mexican cold high plateau conditions. Two groups of gastrointestinal parasitic nematode naturally infected sheep, were randomly selected and located into two free-gastrointestinal nematode larvae paddocks. Group 1 received once a week a supplement containing D. flagrans chlamydospores mixed with oats and molasses. Group 2 received a similar supplement without any fungal material. After 5 months grazing animals were discarded from the experiment and two groups of free-nematode "tracer" sheep were located into the same paddocks to collect larvae from the contaminated pastures. Animals were slaughtered and necropsied and the nematodes were obtained and counted. A screening of the number of gastrointestinal nematode larvae present on the grass was performed and compared between the two grazing areas. The results showed 56% reduction in the Ostertagia (Teladorsagia) circumcincta and 94% reduction in the Nematodirus sp. population of the "tracer" sheep who grazed on the D. flagrans-treated sheep area, compared to the nematode population in animals grazed on the non-treated area. The results of the number of larvae on the grazing pastures showed a 51.1% reduction for H. contortus, and 100% for Cooperia sp. in the area with fungi. In the case of Trichostrongylus sp. no reduction was observed, when compared to the control group. PMID- 17135539 TI - New findings on anaplasmosis caused by infection with Anaplasma phagocytophilum. AB - Ixodes ricinus (I. ricinus) is one of the vectors of Anaplasma phagocytophilum (A. phagocytophilum) in Europe, in which rates of infection range from 1.9% to 34%. In 1998, human granulocytic ehrlichiosis-like (HGE-like) Ehrlichia DNA was detected in Italy, by PCR technique in one I. ricinus nymph out of 55 ticks that were examined. In 1996, 6.3% of 310 human sera in high-risk subjects from Italy were found positive for antibodies to Ehrlichia phagocytophila (E. phagocytophila). In the same year, the authors reported the first case of equine granulocytic ehrlichiosis. In 1997, only 2 out of 563 equine blood samples examined were found positive for antibodies to E. phagocytophila in the Latium region. In 1998, serological positivity was not observed in 14 symptomatic race horses. In 2002, a symptomatic horse living in Rome was found positive for Ehrlichia equi (E. equi) antibodies, as confirmed by PCR. E. equi was also demonstrated in horses by detection of specific antibodies from two asymptomatic ponies. We tested 128 sera from sheep in different flocks, and antibodies to E. phagocytophila were detected in 17 sera (13.3%) of these sheep. From 2000 to 2004, 147 dog sera were tested for antibodies against A. phagocytophilum, and 7 of these sera were positive (4.8%). These data confirm the presence of the infection in human, domestic animals, and pets in Italy. Studies are under way to correlate the distributions of the disease and tick vector, I. ricinus. PMID- 17135540 TI - Molecular diagnosis of granulocytic anaplasmosis and infectious cyclic thrombocytopenia by PCR-RFLP. AB - Anaplasma phagocytophilum (A. phagocytophilum, formerly Ehrlichia phagocytophila) is a tick-borne pathogen responsible for tick-borne fever in ruminants, equine granulocytic ehrlichiosis (EGE) in horses, canine granulocytic ehrlichiosis (CGE) in dogs, and for human granulocytic ehrlichiosis (HGE). Human cases have been registered in many countries with a broad range of symptoms and pathogenicity. This article focused on Sardinia as the prevalence in humans was almost seven times higher than in the rest of Italy. To evaluate the risk, blood samples were collected from dogs and horses on the island. Genomic DNA was extracted from the buffy coat and amplified by heminested polymerase chain reaction (PCR) using the groEL gene primers. The first PCR reaction amplified a 624-bp fragment for both A. phagocytophilum and A. platys while the second PCR reaction amplified 573-bp and 515-bp fragments for the above two pathogens, respectively. Six A. phagocytophilum samples were PCR positive (3 dogs and 3 horses) while another dog was A. platys PCR positive. A phylogenetic analysis was conducted with A. phagocytophilum sequences in GenBank from the United States, Slovenia, Switzerland, Germany, UK, Austria, and Czech Republic. Surprisingly, the related phylogenetic tree showed that the Sardinian isolates were closer to the American isolates, which were showing highest mortality rates than from the other two European lineages. PMID- 17135541 TI - Development of an immunosensor for the diagnosis of bovine anaplasmosis. AB - An optical immunosensor based in major surface protein 5 (MSP5) of Anaplasma marginale was developed towards detection of anti-Anaplasma sp. antibodies in acute infection as well as in vaccinated cattle. This study was performed using recombinant MSP5 covalently immobilised in controlled pore glass (CPG) beads to detect anti-MSP5 antibodies in serum samples. The quantification is based on the measurement of the Cy5 fluorescence of the detection antibody, anti bovine IgG, after reaction with serum. Sera were collected in enzootic and tick-free regions of Argentina. The immunosensor showed a detection range of 1.2 g/ml to 48 g/ml of antibody in sera, with a sensitivity of 93% and a specificity of 70%. The optical immunosensor developed is suitable for quantification of antibodies in sera of naturally or experimentally infected animals. PMID- 17135542 TI - Identification of common antigens in Babesia bovis, B. bigemina, and B. divergens. AB - Bovine babesiosis, caused by Babesia bovis, B. bigemina, and B. divergens, is a significant impediment to livestock production in countries with tropical/subtropical and temperate climates. Previous studies conducted on the immunoprophylaxis against the disease and diagnosis of these parasites has demonstrated the presence of similar antigens. The objective of this article was to identify and partially characterize antigens conserved among these three species. Immunochemical analysis using sera from cattle immunized individually with antigens from these three Babesia species revealed a number of antigens recognized by heterologous antisera. Cross-reactions were more evident in sera from cattle immunized with B. bovis/B. bigemina which recognized several antigens (15 kDa to >200 kDa) in B. divergens. Immunoscreening of a B. divergens cDNA library with bovine serum to B. bigemina allowed the isolation of five clones and DNA sequencing of plasmid BdJF5 showed a 680 bp cDNA insert. Basic Local Alignment Search Tool (BLAST) analysis of the predicted amino acid sequence revealed 47% identity with a protein identified as alphaNAC. Serum from mice immunized with a recombinant Glutathione S-Transferase-BdJF5 fusion protein immunoprecipitated a 20 kDa B. bovis antigen. However, 30 kDa and 18 kDa antigens were immunoprecipitated from B. divergens and immunoblotting analysis revealed the recognition of a 35 kDa B. bigemina antigen. An indirect fluorescence antibody assay on merozoites showed strong reaction with B. divergens and weak recognition of B. bovis and B. bigemina. Despite the existent antigenic polymorphism among the Babesia spp., these results demonstrated that common antigens occur between European B. divergens and Mexican B. bovis/B. bigemina. PMID- 17135543 TI - Evaluation of cattle inoculated with Babesia bovis clones adhesive in vitro to bovine brain endothelial cells. AB - A comparative assessment of the virulence of Babesia bovis clones that adhere or not to bovine brain endothelial cells was done using two clones of B. bovis: (1) a clone phenotypically characterized as virulent (2F8) and (2) a clone of reduced virulence (RAD). Of these subpopulations, we selected those that had adhesive characteristics (a) or nonadhesive characteristics (na) in cultured endothelial cells. Twenty Holstein cattle, 12 months of age or older, were used in this study, and these cattle were randomly assigned to five groups of four animals each. The clones and their respective subpopulations were inoculated via intramuscular injection at a 0.5 x 10(7) infected erythrocyte dosage. Group A was inoculated with aRAD, group B with naRAD, group C with a2F8, group D with na2F8, and group E remained as a control. All inoculated animals showed a decrease in the packed cell volume (PCV), with group D showing the largest decrease (39.53%) and longest time (7 days) with rectal temperature above 39.5 degrees C. Babesia was observed in stained blood smears from only six cattle. While the four parasite subpopulations were pathogenic, significant differences were not noted among them, despite that the subpopulations considered to be virulent caused the greatest reduction in PCV per individual. PMID- 17135544 TI - Bovine babesiosis live vaccine production: use of gamma irradiation on the substrate. AB - Gamma irradiation on bovine serum and red blood cells (RBC) allows proliferation and growth of in vitro-cultured Babesia sp., and has potential application to inactivate contaminating viruses and bacteria from the substrate. Gamma irradiation with 25 kGy in a source of (60)Co was able to inactivate infectious bovine rinotracheitis (IBR) and bovine viral diarrhea (BVD) viruses in artificially contaminated serum; besides, bacteria were also eliminated. In vitro culture of Babesia bovis (B. bovis) in modified substrate, by adding irradiated serum with (60)Co at 25 kGy was propagated from 24-well culture plates to 225 cm(2) tissue culture flasks, and percentages of parasitized erythrocytes (PPE) from 2.4% to 8.8% were obtained. Infected RBC adapted to Irrad S were transferred to the irradiated substrate in vitro culture system, by using serum irradiated at 25 kGy and RBC from 10 to 70 Gy. The PPE ranged from 3.1 to 11. Culture of Babesia bigemina (B. bigemina) was established with Irrad S (25 kGy); its propagation was achieved in tissue culture flasks reaching PPE from 0.5 to 4.3 with no statistical difference (P > 0.05) when compared to the nonirradiated control culture (1.2-4.8). B. bigemina-infected RBCs were transferred to the modified culture system by adding irradiated serum and RBC (25 kGy and 70 Gy, respectively). PPE obtained in culture flasks were from 0.8 to 4.2. The results indicate that gamma irradiation is a suitable method to inactivate potential viral contamination and eliminate bacteria from bovine serum, to produce a live attenuated vaccine through the in vitro culture. PMID- 17135545 TI - Comparative genomics of three strains of Ehrlichia ruminantium: a review. AB - The tick-borne Rickettsiale Ehrlichia ruminantium (E. ruminantium) is the causative agent of heartwater in Africa and the Caribbean. Heartwater, responsible for major losses on livestock in Africa represents also a threat for the American mainland. Three complete genomes corresponding to two different groups of differing phenotypes, Gardel and Welgevonden, have been recently described. One genome (Erga) represents the Gardel group from Guadeloupe Island and two genomes (Erwo and Erwe) belong to the Welgevonden group. Erwo, isolated in South Africa, is the parental strain of Erwe, which was maintained for 18 years in Guadeloupe under different culture conditions than Erwo. The three strains display genomes of differing sizes with 1,499,920 bp, 1,512,977 bp, and 1,516,355 bp for Erga, Erwe, and Erwo, respectively. Gene sequences and order are highly conserved between the three strains, although several gene truncations could be pinpointed, most of them occurring within three regions of accumulated differences (RAD). E. ruminantium displays a strong leading/lagging compositional bias inducing a strand-specific codon usage. Finally, a striking feature of E. ruminantium is the presence of long intergenic regions containing tandem repeats. These repeats are at the origin of an active process, specific to E. ruminantium, of genome expansion/contraction based on the addition or removal of tandem units. PMID- 17135546 TI - Emerging tick-borne disease in African vipers caused by a Cowdria-like organism. AB - Heartwater is a tick-borne infectious disease caused by the rickettsial organism Cowdria ruminantium, currently Ehrlichia ruminantium. It poses an imminent threat to the Western Hemisphere, where it could cause mortality in cattle and other ruminant livestock in excess of 70%. It has been reported in the Caribbean; and its vector, Amblyomma sparsum, has been found on imported African spurred tortoises (Geochelone sulcata) and leopard tortoises (Geochelone pardalis) in southern Florida in the United States, leading to an importation ban on these reptiles. Symptoms have not been previously reported in reptiles. Here, we report peracute and acute deaths in African vipers imported from Africa through Florida. Signs included vomiting mucoid fluid, diarrhea, emaciation, convulsions, and death. Postmortem showed few gross lesions. The most consistent peracute and acute lesions were the pulmonary lesions and pericarditis with considerable bloody fluid in the pericardial sac (hydropericardium). These lesions strongly resembled the lesions of heartwater and a coccobacillus of less than 1-micron diameter was isolated in viper cell culture. The outbreak was brought to a halt by tick control and treatment of all exposed snakes with tetracycline. This isolation, tetracycline sensitivity, clinical signs, preliminary results with polymerase chain reaction of pCS20 ORF, and the viper preference of the disease may indicate a Cowdria-related attenuated species that has adapted to infect reptiles or an emerging new form of this group of microbes. PMID- 17135547 TI - Identification and characterization of merozoite antigens of a Theileria species highly pathogenic for small ruminants in China. AB - A new pathogenic Theileria species transmitted by Haemaphysalis qinghaiensis was identified in the Northwestern part of China and was shown to be highly pathogenic for small ruminants. The present article aimed at identifying merozoite antigens that might be suitable for developing diagnostic methods and designing a potential vaccine. Absence of other theilerial or babesial infections was confirmed by reverse line blot in all antigen samples used. Extensive Western blot analyses using serum from infected and noninfected animals led to the identification of four potential merozoite immunoreactive proteins at different molecular weights. Further protein characterization using peptide mass mapping by matrix-assisted laser desorption/ionization (MALDI) followed by database searching resulted in two significant hits that identified two proteins of parasite origin, one homologous to a possible MO25-family protein from Cryptosporidium parvum and the other with an HSP70 from Theileria annulata. Another protein was also identified as a parasite protein but without significant homology. Immunization of rabbits with selected proteins produced antisera that reacted specifically on Western blots with merozoite antigens of the corresponding sizes. This article represents the first identification and characterization of potential antigenic proteins of Theileria sp. (China) for veterinary purposes. PMID- 17135548 TI - Purification of macroschizonts of a Sudanese isolate of Theileria lestoquardi (T. lestoquardi [Atbara]). AB - Research on malignant theileriosis is affected by the limited access to biological materials required for studies aiming at controlling the disease through the establishment of diagnostic tools and vaccines. The main aims of this work were to isolate, establish, and characterize a Theileria lestoquardi infected cell culture (line) as a source of biological material and to generate a schizont cDNA library for further studies aiming at the identification of antigenic proteins. The T. lestoquardi isolate used originated from a sheep showing typical signs of malignant theileriosis in Atbara town in northern Sudan, and was maintained as an infected cell culture. A high-quality representative schizont cDNA library was established by isolating and purifying the schizonts using a nocodazole/aerolysin protocol followed by Percoll gradient ultracentrifugation. As a parameter to assess the quality of the schizont library, a provisional estimation of the percentage of recombinant phage clones originating from T. lestoquardi (Atbara) was undertaken. Ten clones with inserts ranging in size between 600 and 1200 bp were selected randomly, sequenced, and subjected to BLAST similarity searches. As 6 of the 10 sequenced clones showed similarities to T. parva, T. annulata, and other apicomplexan genes, it was concluded that the majority of the library phage clones originated from the parasite and not from host cell transcripts. The cDNA library will be used for screening of antigenic proteins using sera from infected sheep. PMID- 17135549 TI - Identification of potential antigenic proteins of Theileria lestoquardi. AB - A PCR strategy was used to identify potential antigenic proteins of T. lestoquardi suitable for the development of an ELISA by searching for homologous proteins previously identified in other theilierial parasites to be antigenic. PMID- 17135550 TI - The host responses in sheep artificially infected with Theileria sp. (China). AB - Studies on host responses in sheep artificially infected with Theileria sp. (China) were discussed and summarized mainly on typical high fever periods, merozoits and schizoon observation, antibody response. PMID- 17135551 TI - Identification of homologous genes of T. annulata proteins in the genome of Theileria sp. (China). AB - Homologues to previously described Theileria (T.) annulata genes (T. annulata surface protein [TaSP], putative T. annulata membrane protein [TaD]) were successfully amplified by polymerase chain reaction (PCR) from Theileria sp. (China) merozoite cDNA, with 88% identity to TaD; TcSP partial cDNA, 94% identity to TaSP. Moreover, homologues to a secretory protein of T. annulata (TaSE), with a sequence identity of 99% on the cDNA level (TcSE partial cDNA) and to a potential membrane protein of T. lestoquardi (Clone-5), with a sequence identity of 100% on the genomic level (Tc Clone-5) but lacking an intron at positions 1894 1928 were identified. PMID- 17135552 TI - Epidemiology of Theileria annulata infection of dairy cattle in the Sudan using molecular techniques. AB - This study provides the first epidemiological data regarding T. annulata infection of diary cattle in Sudan using a combination of routine microscopic examination and two molecular techniques, PCR and reverse line blot (RLB). PMID- 17135553 TI - Investigation of MAP kinase activation in Theileria-infected cell lines. AB - A study on different Theileria annulata and Theileria parva infected cell lines was performed in order to evaluate the general relevance of the MAP-kinase activation status for Theileria- mediated transformation. PMID- 17135554 TI - Dermal mast cell counts in F2 Holstein x Gir crossbred cattle artificially infested with the tick Boophilus microplus (Acari: Ixodidae). AB - The role of dermal mast cells (DMC) in the host resistance to ticks has been studied but it is not totally explained yet. Studies have proposed that zebuine cattle breeds, known as highly resistant to ticks, have more DMC than taurine breeds. In the present study, we compared the number of adult female ticks Boophilus microplus and the mast cells' countings in the skin of F(2) crossbred Gir x Holstein cattle, before and after tick infestation. F(2) crossbred cattle (n = 148) were divided into seven groups and artificially infested with 1.0 x 10(4) B. microplus larvae and, 21 days afterwards, adult female-fed ticks attached to the skin were counted. Skin biopsies were taken and examined under light microscopy with a square-lined ocular reticulum in a total area of 0.0625 mm(2) in both the superficial and deep dermis. Results demonstrated that infested F(2) crossbred cattle acquired resistance against the cattle-tick B. microplus probably associated to an increase in the dermal mast cell number. It is concluded that the tick infestation may lead to an environmental modification in the dermis of parasitized hosts due to the massive migration of mast cells or their local proliferation. PMID- 17135555 TI - Detection of Hepatozoon canis in stray dogs and cats in Bangkok, Thailand. AB - A rapidly increasing stray animal population in Bangkok has caused concern regarding transmission of vector-borne and zoonotic diseases. The purpose of this study was to determine if stray animals in Bangkok are a potential reservoir of Hepatozoon, a genus of tick-borne parasites that has received little attention in Thailand. Blood samples were collected from stray companion animals near monasteries in 42 Bangkok metropolitan districts. Both dogs and cats were sampled from 26 districts, dogs alone from 4 districts and cats alone from 12 districts. Samples were collected from a total of 308 dogs and 300 cats. Light microscopy and an 18 S rRNA gene-based PCR assay were used to test these samples for evidence of Hepatozoon infection. Gamonts were observed in blood smears for 2.6% of dogs and 0.7% of cats by microscopy. The PCR assay detected Hepatozoon in buffy coats from 11.4% of dogs and 32.3% of cats tested. The prevalence of infection was the same between male and female dogs or cats, and PCR-positive dogs and cats were found in 36.6% and 36.8% of the districts surveyed, respectively. There was an association between the percentages of PCR-positive dogs and cats in districts where both host species were sampled. Sequences of representative amplicons were closest to those reported for H. canis. These results represent the first molecular confirmation that H. canis is indigenous to Thailand. The unexpectedly high prevalence of Hepatozoon among stray cats indicates that their role in the epizootiology of hepatozoonosis should be investigated. PMID- 17135556 TI - Renitelo cattle dermatophilosis and PCR-RFLP analysis of MHC gene. AB - Renitelo breed is a cattle breed created at Kianjasoa station (Madagascar) by a triple crossing Malagasy Zebu x Limousine x Afrikander. This breed besides many valuable advantages, such as rapid growth and drought power, presents a huge disadvantage which is sensitivity to skin disease, dermatophilosis, previously known as streptotrichosis. This disease caused by Dermatophilus congolensis is one of the major threats for the population of Renitelo cattle. An allele of MHC gene has been shown to be dramatically associated to hypersensitivity to the disease in other cattle breed. To bring further information to tick borne disease clinical survey, mainly dermatophilosis, we wanted to verify if such allele could be found in this breed. Renitelo cattle included in this study were chosen for the presence of dermatophilosis lesions in more or less severe form (N = 17). These animals were blood sampled and a genetic analysis on the MHC gene BoLA-DRB3 was performed, by PCR amplification using BOD 31 & BOD 32 primers. Amplified products were analyzed by RFLP using enzymes. Restriction band profiles were characterized according to previously defined patterns. Three cows out of the 17 cattle analyzed for MHC gene presented the hypersensitive allele FDA. Two out of the three hypersensitive cows were pure breed while one was half breed. All the cows presented dermatophilosis lesions at least during rainy season but one of them particularly suffered from severe lesions covering all its body and died of the illness. This study shows that hypersensitivity allele found in other bovine breeds can be found in Renitelo breed. This result seemed to suggest that this characterization could be utilized in breeding program for this breed. PMID- 17135557 TI - Polyphasic taxonomy. AB - Several organisms from a number of prokaryotic and eukaryotic groups have presented problems for systematists for a long time. Both phenotypic and genotypic methods for sorting out these relationships have been employed. There are limitations with each method when taken alone. Since the purpose of systematics is to determine the correct genealogical relationships among biological organisms, it is necessary to use all available means to arrive at consensus associations, and polyphasic taxonomy, which takes into consideration both methods, is a rational approach. In this short article, we provide a number of examples where polyphasic taxonomy is serving as the means of arriving at the desired consensus. PMID- 17135558 TI - Phylogenetic position of small-ruminant infecting piroplasms. AB - Theileria and Babesia are tick-transmitted protozoa that cause great economical losses in livestock. Recently, interest has risen in sheep-infecting piroplasms and a number of previously unidentified pathogens were described, particularly in China. To address the phylogenetic relationship of Theileria and Babesia species infecting sheep, the complete sequences of the 18 S small subunit ribosomal RNA genes of a panel of piroplasm isolates, including T. lestoquardi, T. ovis, T. separata, B. ovis, B. motasi, B. crassa, and several novel species, were compared. The classification based on the established phylogenetic tree corresponded with traditional systematics and revealed that sheep/goat piroplasm species are of a polyphyletic origin. In addition, these studies revealed the existence of at least two novel sheep/goat piroplasm species, designated Theileria sp. (China 1) and Theileria sp. (China 2). PMID- 17135559 TI - Phylogenetics of Theileria species in small ruminants. AB - Our study is based on the collection of blood and ticks from sheep in Iran and Italy. Polymerase chain reaction (PCR) testing was performed to target the 18S rRNA gene and RLB was performed using previously published probes. In Italy and Iran 78.7% and 76.0% of the sheep were PCR positive, which after sequencing and RLB showed that they were Theileria ovis and Theileria lestoquardi, respectively. Phylogenetic analysis was performed using the Clustal W multiple sequence alignment program and our sequences were compared with more than 50 others already published in the EMBL database. Our T. lestoquardi sequences linked with other T. lestoquardi sequences from Iran, Tanzania, and Sudan and Theileria annulata showed the importance of having species-specific probes between these two species. However, distinctive clades were found between T. lestoquardi ticks and those found in sheep blood. Italian T. ovis seemed to be closer to Theileria spp. from Namibia and Iran than with other T. ovis from Spain, Turkey, Tanzania, and Sudan adding some information to the controversy about this species. However, some confusion was found on the existing database where the location of pathogens, years, and species names was inaccurate and when available sequences were not always appropriately used. This article will discuss our results and some comparisons with other phylogenetic approaches. PMID- 17135560 TI - Molecular sequence evidence for the reclassification of some Babesia species. AB - Taxonomic characterization of organisms in the genera Theileria and Babesia was originally based on observations of morphology and certain general phenotypic characteristics, which enabled many parasites to be unequivocally assigned to a particular genus. However, application of molecular genetic techniques, such as the polymerase chain reaction (PCR) for gene amplification, and DNA sequencing, have revealed gross inconsistencies in the assignation of some parasite genetic variants, particularly those of the B. gibsoni and B. microti complexes, to the genus Babesia. These variants cannot be assigned, on the basis of sequence information and phylogenetic analysis, to either of the genera Theileria and Babesia. The gene for which most sequence information is available for phylogenetic analysis is the small subunit ribosomal RNA (srRNA) gene. This gene allows clear distinction of the genera Theileria and Babesia (sensu stricto) and reveals that many "Babesia" variants are phylogenetically distinct from both genera. This distinction is confirmed, for some of the variants, by beta-tubulin sequence data, suggesting that the organisms should be renamed and reclassified. PMID- 17135561 TI - Phylogenetic and biologic evidence that Babesia divergens is not endemic in the United States. AB - The causative agent of human babesiosis in a Kentucky case, which was first identified as Babesia divergens, is identical to a parasite of eastern cottontail rabbits on Nantucket Island, Massachusetts based on piroplasm size, morphology, and ribosomal RNA sequence analysis. Studies showing differential infectivity for cattle, host erythrocyte specificity in vitro, parasite size and morphology in vitro, and ribosomal RNA sequences clearly demonstrate that the parasite from the rabbit (conspecific with the human Kentucky agent) is not the same organism as B. divergens. PMID- 17135562 TI - An Internet portal dedicated to pig production and wild suids in the tropics: PigTrop web site http://pigtrop.cirad.fr. AB - Considering that a wide access to updated and relevant data is a key point for livestock development and research improvement in tropics, The PigTrop web site (http://pigtrop.cirad.fr) is dedicated to pig production and pork commodity chains in developing countries. It mainly addresses stakeholders involved in the pig commodity chain, but also researchers, students, or development agencies with an interest in tropical pig breeding. It is run by the French Agricultural Research Centre for International Development (CIRAD). PMID- 17135563 TI - Local epidemiosurveillance in swine diseases in Northern Vietnam: description and preliminary results. AB - The REMAPORC is an epidemiosurveillance network in swine diseases and an organizational model for local veterinary services in one district of Northern Vietnam. A strong concern was done on quality of the sanitary information chain from field and feedback to local agents. Based on 4,000 declarations provided by veterinarians and animal health workers involved, preliminary results highlighted the major incidence of porcine respiratory disease complex; digestive affections in piglets, and reproductive disorders in newly raised exotic sows have been also noticed. PMID- 17135564 TI - A study of edema disease in pigs in Vietnam with particular reference to the use of autovaccine for the prevention of disease. AB - Edema disease caused by Escherichia coli is one of the most common diseases in postweaning piglets throughout Vietnam. Verotoxigenic E. coli (VTEC) was isolated from 197 of 261 samples (75.5%). All isolates were confirmed by basic biochemical tests and carbohydrate fermentation characteristics. Of these, 70.1% of isolates are hemolytic, 45% isolates belonged to serotypes O149:K91, possessed the VT2e gene, and was the most predominant VTEC pathotype associated with edema disease in pigs. Serogroup O139 accounted for 30% of the isolates, followed by serogroup O138 and O141 (25%). In addition to VT2e gene, the ST (72.7%) and LT (52.7%) genes were also recognized. A total of 10 representative isolates were subjected to toxigenicity testing by intraperitoneal injection in mice and experimental infection in pigs. It was shown that 100% of the mice were killed 17-24 h post injection (p.i.). All pigs experimentally infected with challenge strains and developed typical symptoms of edema disease 36-72 h p.i. A multivalent killed whole-cells vaccine containing aluminum hydroxide was prepared from 5 VTEC strains. The vaccine was 100% safe when administered by the intramuscular route into the pigs. A field trial for over 100,000 pigs (21-90 days old) showed that vaccinated pigs were protected against edema disease at a level of 90% compared to 100% of pigs from unvaccinated groups. PMID- 17135565 TI - Preliminary analysis of tetracycline residues in marketed pork in Hanoi, Vietnam. AB - A cross-sectional survey was designed to investigate the proportion of tetracycline residues in marketed pork in suburb and urban districts in Hanoi. A total of 290 raw muscle samples were randomly collected from open markets in these districts. The samples were qualitatively screened for tetracycline residues using the agar inhibition test, and Bacillus cereus (ATCC 11778) as the reference strain. The inconclusive samples were then analyzed using high performance liquid chromatography (HPLC). The positive samples from either test were defined as positive results. Overall, 5.5% of all collected samples were positive for tetracycline residues. The proportion of positive samples from shops in suburb districts was significantly (P < 0.05) different from those collected from shops in urban districts. So, the factor of region was identified as a risk factor of tetracycline residue proportion in raw pork with an odds ratio (OR) of 4.03 (95% CI = 1.12, 14.45). For the other factors, such as season, type of shop, type of abattoir, origin of meat, etc., the difference in proportion of positive samples within each factor was substantial but not statistically significant. These factors were identified as nonrisk factors. Such a high proportion may pose a potential hazard to public health, particularly since they might induce drug resistance of pathogenic micro-organisms. PMID- 17135566 TI - Antimicrobial resistance phenotypes of ETEC isolates from piglets with diarrhea in North Vietnam. AB - Both disk diffusion and broth micro-dilution assays were employed to determine the level of resistance in Enterotoxigenic Escherichia coli (ETEC) isolates (n = 170) obtained from preweaning piglet colibacillosis from the two different pig production systems (commercial piggeries and small holder farmers) in Vietnam. Overall, tetracycline, streptomycin, amoxicillin, trimethoprim/sulfamethoxazole, and chloramphenicol showed markedly higher rates of resistance. Both apramycin and ceftiofur are active against all ETEC isolates. These antimicrobials could be recommended as the drugs of choice for the treatment of E. coli infections in young pigs in North Vietnam. Resistance to third-generation cephalosporin (ceftiofur, ceftazidime, and cefoxitin) was not observed in Vietnamese ETEC isolates. Multiple resistances to greater than three antimicrobials were widely distributed (approximately 79.4%). PMID- 17135567 TI - Influence of different diets on growth performance, meat quality, and disease resistance in pig crossbreeds (PIE x MC-local) and PIE (LW x MC-local). AB - The present study evaluated the effect of different dietary formulations on the growth rate in pigs and their resistance to infection with hog cholera. Results indicate that growth rates can be enhanced by certain formulations and that there is a correlation between this increased growth rate and increased resistance to infection with hog cholera. PMID- 17135568 TI - Intracerebral hemorrhages in CADASIL. AB - Intracerebral hemorrhage (ICH) has been described only sporadically for patients with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). However, cerebral microbleeds (CMBs) were found in 31% to 69% of the patients with CADASIL, and this predicted an increased risk of ICH. In this study, the authors found that 25% of the symptomatic patients with CADASIL had ICHs, and their development was closely related to the number of CMBs. PMID- 17135569 TI - Genome-wide prediction of conserved and nonconserved enhancers by histone acetylation patterns. AB - Comparative genomic studies have been useful in identifying transcriptional regulatory elements in higher eukaryotic genomes, but many important regulatory elements cannot be detected by such analyses due to evolutionary variations and alignment tool limitations. Therefore, in this study we exploit the highly conserved nature of epigenetic modifications to identify potential transcriptional enhancers. By using a high-resolution genome-wide mapping technique, which combines the chromatin immunoprecipitation and serial analysis of gene expression assays, we have recently determined the distribution of lysine 9/14-diacetylated histone H3 in human T cells. We showed the existence of 46,813 regions with clusters of histone acetylation, termed histone acetylation islands, some of which correspond to known transcriptional regulatory elements. In the present study, we find that 4679 sequences conserved between human and pufferfish coincide with histone acetylation islands, and random sampling shows that 33% (13/39) of these can function as transcriptional enhancers in human Jurkat T cells. In addition, by comparing the human histone acetylation island sequences with mouse genome sequences, we find that despite the conservation of many of these regions between these species, 21,855 of these sequences are not conserved. Furthermore, we demonstrate that about 50% (26/51) of these nonconserved sequences have enhancer activity in Jurkat cells, and that many of the orthologous mouse sequences also have enhancer activity in addition to conserved epigenetic modification patterns in mouse T-cell chromatin. Therefore, by combining epigenetic modification and sequence data, we have established a novel genome-wide method for identifying regulatory elements not discernable by comparative genomics alone. PMID- 17135570 TI - Localization of a long-range cis-regulatory element of IL13 by allelic transcript ratio mapping. AB - It appears that, for many genes, the two alleles possessed by an individual may produce different amounts of transcript. When such allelic differences in transcription are observed for some individuals but not others, a plausible explanation is genetic variation in the cis-acting elements that regulate the gene in question. Here we describe a novel analytical approach that uses such observations, combined with genotyping data from the HapMap project, to define the genomic location of cis-acting regulatory elements. When applied to the human 5q31 chromosomal region, where complex regulatory mechanisms are known to exist, we demonstrate the sensitivity of this approach by locating a highly significant cis-regulatory element operating on IL13 at long range from a position 250 kb upstream from the gene (P = 2 x 10(-6)). As this method is unaffected by other sources of variation, such as environmental and trans-acting genetic factors, it provides a tractable approach for dissecting the complexities of genetic variation in gene regulation. PMID- 17135571 TI - Large-scale production of SAGE libraries from microdissected tissues, flow-sorted cells, and cell lines. AB - We describe the details of a serial analysis of gene expression (SAGE) library construction and analysis platform that has enabled the generation of >298 high quality SAGE libraries and >30 million SAGE tags primarily from sub-microgram amounts of total RNA purified from samples acquired by microdissection. Several RNA isolation methods were used to handle the diversity of samples processed, and various measures were applied to minimize ditag PCR carryover contamination. Modifications in the SAGE protocol resulted in improved cloning and DNA sequencing efficiencies. Bioinformatic measures to automatically assess DNA sequencing results were implemented to analyze the integrity of ditag structure, linker or cross-species ditag contamination, and yield of high-quality tags per sequence read. Our analysis of singleton tag errors resulted in a method for correcting such errors to statistically determine tag accuracy. From the libraries generated, we produced an essentially complete mapping of reliable 21 base-pair tags to the mouse reference genome sequence for a meta-library of approximately 5 million tags. Our analyses led us to reject the commonly held notion that duplicate ditags are artifacts. Rather than the usual practice of discarding such tags, we conclude that they should be retained to avoid introducing bias into the results and thereby maintain the quantitative nature of the data, which is a major theoretical advantage of SAGE as a tool for global transcriptional profiling. PMID- 17135572 TI - Hydrogen peroxide stimulates macrophages and monocytes to actively release HMGB1. AB - High mobility group box 1 (HMGB1) can be actively secreted by macrophages/monocytes in response to exogenous and endogenous inflammatory stimuli (such as bacterial endotoxin, TNF-alpha, IL-1, and IFN-gamma) or passively released by necrotic cells and mediates innate and adaptive inflammatory responses to infection and injury. Here, we demonstrated that a reactive oxygen species, hydrogen peroxide (H(2)O(2)), induces active and passive HMGB1 release from macrophage and monocyte cultures in a time- and dose-dependent manner. At nontoxic doses (e.g., 0.0125-0.125 mM), H(2)O(2) induced HMGB1 cytoplasmic translocation and active release within 3-24 h. At higher concentrations (e.g., 0.25 mM), however, H(2)O(2) exhibited cytotoxicity to macrophage and monocyte cell cultures and consequently, triggered active and passive HMGB1 release. In addition, H(2)O(2) stimulated potential interaction of HMGB1 with a nuclear export factor, chromosome region maintenance (CRM1), in macrophage/monocyte cultures. Inhibitors specific for the JNK (SP600125) and MEK (PD98059), but not p38 MAPK (SB203580), abrogated H(2)O(2)-induced, active HMGB1 release. Together, these data establish an important role for oxidative stress in inducing active HMGB1 release, potentially through a MAPK- and CRM1-dependent mechanism. PMID- 17135573 TI - The CD14+ CD16+ blood monocytes: their role in infection and inflammation. AB - Blood monocyte subpopulations have been defined in man initially, and the two major types of monocytes are the CD14++ CD16- and the CD14+ CD16+ monocytes. These cells have been shown to exhibit distinct phenotype and function, and the CD14+ CD16+ were labeled proinflammatory based on higher expression of proinflammatory cytokines and higher potency in antigen presentation. The current review describes these properties, including the relationship to dendritic cells, and summarizes the host of publications about CD14+ CD16+ monocytes in inflammation and infectious disease in man, all of which suggest a crucial role of these cells in the disease processes. The review also covers the more recent description of homologues of these cells in other model species, which is expected to better define the role of monocyte subsets in disease. PMID- 17135574 TI - Relevance of DC-SIGN in DC-induced T cell proliferation. AB - The role of dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN) in DC-T cell communication was assessed by analyzing the effect of DC-SIGN-blocking mAb in MLR. The results show that the degree of inhibition by DC-SIGN and LFA-1 mAb depends on the magnitude of the MLR and the maturation status of the DC. Addition of DC-SIGN mAb at several time-points during MLR showed that DC-SIGN is involved early on in DC-T cell contacts. This initial role is masked by strong adhesive and costimulatory mechanisms, indicating a short-lived effect of DC-SIGN in DC-T cell interactions. To examine this concept in more detail, the percentage of PBL capable of binding DC-SIGN was determined. Analysis of several donors revealed that 1-20% PBL bind to beads coated with recombinant DC-SIGN, and the DC-SIGN binding cells comprised all major cell subsets found in blood. PBL isolated from a donor with high DC-SIGN-binding capacity were more prone to blocking by DC-SIGN mAb in MLR than PBL from a donor with low DC-SIGN-binding capacity. This study indicates an initial and transient role for DC-SIGN in T cell proliferation, which becomes apparent when T cell proliferation is low and when the percentage of DC-SIGN binding PBL is high. PMID- 17135575 TI - Prostaglandin I2 analogs inhibit Th1 and Th2 effector cytokine production by CD4 T cells. AB - An anti-inflammatory effect of PGI(2) has been suggested by increased inflammation in mice that are deficient in the PGI(2) receptor (IP) or in respiratory syncytial viral- or OVA-induced CD4 T cell-associated responses. To determine the mechanism of the anti-inflammatory effect, we hypothesized that PGI(2) analogs inhibit CD4 T cell effector cytokine production. To test this hypothesis, we activated purified CD4 T cells with anti-CD3 and anti-CD28 antibodies under Th1 and Th2 polarizing conditions for 4 days and restimulated the T cells with anti-CD3 in the presence of PGI(2) analogs for 2 days. We found that PGI(2) analogs (cicaprost and iloprost) inhibited the production of Th1 cytokines (IFN-gamma) and Th2 cytokines (IL-4, IL-10, and IL-13) in a dose dependent pattern. The inhibitory effect was partially dependent on the IP receptor signaling and was correlated with elevated intracellular cAMP and down regulated NF-kappaB activity. Pretreatment of the CD4 T cells with 8 bromoadenosine-3',5'-cyclic monophosphorothioate, Rp-isomer, to inhibit a key signaling molecule in the cAMP pathway, protein kinase A (PKA), attenuated the suppressive effect of PGI(2) analogs significantly, suggesting that PKA, in part, mediates the inhibition of the cytokine production. These data indicate that PGI(2) analogs have an immune-suppressive effect on previously activated and differentiated CD4 T cells in vitro and suggest that PGI(2) may have a similar function in vivo. PMID- 17135576 TI - MCP-1 deficiency causes altered inflammation with impaired skeletal muscle regeneration. AB - We examined the role of MCP-1, a potent chemotactic and activating factor for macrophages, in perfusion, inflammation, and skeletal muscle regeneration post ischemic injury. MCP-1-/- or C57Bl/6J control mice [wild-type (WT)] underwent femoral artery excision (FAE). Muscles were collected for histology, assessment of tissue chemokines, and activity measurements of lactate dehydrogenase (LDH) and myeloperoxidase. In MCP-1-/- mice, restoration of perfusion was delayed, and LDH and fiber size, indicators of muscle regeneration, were decreased. Altered inflammation was observed with increased neutrophil accumulation in MCP-1-/- versus WT mice at Days 1 and 3 (P< or =0.003), whereas fewer macrophages were present in MCP-1-/- mice at Day 3. As necrotic tissue was removed in WT mice, macrophages decreased (Day 7). In contrast, macrophage accumulation in MCP-1-/- was increased in association with residual necrotic tissue and impaired muscle regeneration. Consistent with altered inflammation, neutrophil chemotactic factors (keratinocyte-derived chemokine and macrophage inflammatory protein-2) were increased at Day 1 post-FAE. The macrophage chemotactic factor MCP-5 was increased significantly in WT mice at Day 3 compared with MCP-1-/- mice. However, at post-FAE Day 7, MCP-5 was significantly elevated in MCP-1-/- mice versus WT mice. Addition of exogenous MCP-1 did not induce proliferation in murine myoblasts (C2C12 cells) in vitro. MCP-1 is essential for reperfusion and the successful completion of normal skeletal muscle regeneration after ischemic tissue injury. Impaired muscle regeneration in MCP-1-/- mice suggests an important role for macrophages and MCP-1 in tissue reparative processes. PMID- 17135577 TI - Misunderstanding prescription labels: the genie is out of the bottle. PMID- 17135578 TI - Literacy and misunderstanding prescription drug labels. AB - BACKGROUND: Health literacy has increasingly been viewed as a patient safety issue and may contribute to medication errors. OBJECTIVE: To examine patients' abilities to understand and demonstrate instructions found on container labels of common prescription medications. DESIGN: Cross-sectional study using in-person, structured interviews. SETTING: 3 primary care clinics serving mostly indigent populations in Shreveport, Louisiana; Jackson, Michigan; and Chicago, Illinois. PATIENTS: 395 English-speaking adults waiting to see their providers. MEASUREMENT: Correct understanding of instructions on 5 container labels; demonstration of 1 label's dosage instructions. RESULTS: Correct understanding of the 5 labels ranged from 67.1% to 91.1%. Patients reading at or below the sixth grade level (low literacy) were less able to understand all 5 label instructions. Although 70.7% of patients with low literacy correctly stated the instructions, "Take two tablets by mouth twice daily," only 34.7% could demonstrate the number of pills to be taken daily. After potential confounding variables were controlled for, low (adjusted relative risk, 2.32 [95% CI, 1.26 to 4.28]) and marginal (adjusted relative risk, 1.94 [CI, 1.14 to 3.27]) literacy were significantly associated with misunderstanding. Taking a greater number of prescription medications was also statistically significantly associated with misunderstanding (adjusted relative risk, 2.98 [CI, 1.40 to 6.34] for > or =5 medications). LIMITATIONS: The study sample was at high risk for poor health literacy and outcomes. Most participants were women, and all spoke English. The authors did not examine the association between misunderstanding and medication error or evaluate patients' actual prescription drug-taking behaviors. CONCLUSIONS: Lower literacy and a greater number of prescription medications were independently associated with misunderstanding the instructions on prescription medication labels. PMID- 17135579 TI - Images in clinical medicine. Bisphosphonate-associated contact stomatitis. PMID- 17135580 TI - Pioneers in AIDS care--reflections on the epidemic's early years. PMID- 17135581 TI - HIV-AIDS in Tanzania--realities on the ground. PMID- 17135582 TI - Osteonecrosis of the jaw--do bisphosphonates pose a risk? PMID- 17135583 TI - CD4+ count-guided interruption of antiretroviral treatment. AB - BACKGROUND: Despite declines in morbidity and mortality with the use of combination antiretroviral therapy, its effectiveness is limited by adverse events, problems with adherence, and resistance of the human immunodeficiency virus (HIV). METHODS: We randomly assigned persons infected with HIV who had a CD4+ cell count of more than 350 per cubic millimeter to the continuous use of antiretroviral therapy (the viral suppression group) or the episodic use of antiretroviral therapy (the drug conservation group). Episodic use involved the deferral of therapy until the CD4+ count decreased to less than 250 per cubic millimeter and then the use of therapy until the CD4+ count increased to more than 350 per cubic millimeter. The primary end point was the development of an opportunistic disease or death from any cause. An important secondary end point was major cardiovascular, renal, or hepatic disease. RESULTS: A total of 5472 participants (2720 assigned to drug conservation and 2752 to viral suppression) were followed for an average of 16 months before the protocol was modified for the drug conservation group. At baseline, the median and nadir CD4+ counts were 597 per cubic millimeter and 250 per cubic millimeter, respectively, and 71.7% of participants had plasma HIV RNA levels of 400 copies or less per milliliter. Opportunistic disease or death from any cause occurred in 120 participants (3.3 events per 100 person-years) in the drug conservation group and 47 participants (1.3 per 100 person-years) in the viral suppression group (hazard ratio for the drug conservation group vs. the viral suppression group, 2.6; 95% confidence interval [CI], 1.9 to 3.7; P<0.001). Hazard ratios for death from any cause and for major cardiovascular, renal, and hepatic disease were 1.8 (95% CI, 1.2 to 2.9; P=0.007) and 1.7 (95% CI, 1.1 to 2.5; P=0.009), respectively. Adjustment for the latest CD4+ count and HIV RNA level (as time-updated covariates) reduced the hazard ratio for the primary end point from 2.6 to 1.5 (95% CI, 1.0 to 2.1). CONCLUSIONS: Episodic antiretroviral therapy guided by the CD4+ count, as used in our study, significantly increased the risk of opportunistic disease or death from any cause, as compared with continuous antiretroviral therapy, largely as a consequence of lowering the CD4+ cell count and increasing the viral load. Episodic antiretroviral therapy does not reduce the risk of adverse events that have been associated with antiretroviral therapy. (ClinicalTrials.gov number, NCT00027352 [ClinicalTrials.gov].). PMID- 17135584 TI - A placebo-controlled trial of pioglitazone in subjects with nonalcoholic steatohepatitis. AB - BACKGROUND: No pharmacologic therapy has conclusively proved to be effective for the treatment of nonalcoholic steatohepatitis, which is characterized by insulin resistance, steatosis, and necroinflammation with or without centrilobular fibrosis. Pioglitazone is a thiazolidinedione that ameliorates insulin resistance and improves glucose and lipid metabolism in type 2 diabetes mellitus. METHODS: We randomly assigned 55 patients with impaired glucose tolerance or type 2 diabetes and liver biopsy-confirmed nonalcoholic steatohepatitis to 6 months of treatment with a hypocaloric diet (a reduction of 500 kcal per day in relation to the calculated daily intake required to maintain body weight) plus pioglitazone (45 mg daily) or a hypocaloric diet plus placebo. Before and after treatment, we assessed hepatic histologic features, hepatic fat content by means of magnetic resonance spectroscopy, and glucose turnover during an oral glucose tolerance test ([14C]glucose given with the oral glucose load and [3H]glucose given by intravenous infusion). RESULTS: Diet plus pioglitazone, as compared with diet plus placebo, improved glycemic control and glucose tolerance (P<0.001), normalized liver aminotransferase levels as it decreased plasma aspartate aminotransferase levels (by 40% vs. 21%, P=0.04), decreased alanine aminotransferase levels (by 58% vs. 34%, P<0.001), decreased hepatic fat content (by 54% vs. 0%, P<0.001), and increased hepatic insulin sensitivity (by 48% vs. 14%, P=0.008). Administration of pioglitazone, as compared with placebo, was associated with improvement in histologic findings with regard to steatosis (P=0.003), ballooning necrosis (P=0.02), and inflammation (P=0.008). Subjects in the pioglitazone group had a greater reduction in necroinflammation (85% vs. 38%, P=0.001), but the reduction in fibrosis did not differ significantly from that in the placebo group (P=0.08). Fatigue and mild lower-extremity edema developed in one subject who received pioglitazone; no other adverse events were observed. CONCLUSIONS: In this proof-of-concept study, the administration of pioglitazone led to metabolic and histologic improvement in subjects with nonalcoholic steatohepatitis. Larger controlled trials of longer duration are warranted to assess the long-term clinical benefit of pioglitazone. (ClinicalTrials.gov number, NCT00227110 [ClinicalTrials.gov] .). PMID- 17135585 TI - Financial relationships between institutional review board members and industry. AB - BACKGROUND: Little is known about the nature, extent, and consequences of financial relationships between industry and institutional review board (IRB) members in academic institutions. We surveyed IRB members about such relationships. METHODS: We surveyed a random sample of 893 IRB members at 100 academic institutions (response rate, 67.2%). The questionnaire focused on the financial relationships that the members had with industry (e.g., employment, membership on boards, consulting, receipt of royalties, and paid speaking). RESULTS: We found that 36% of IRB members had had at least one relationship with industry in the past year. Of the respondents, 85.5% said they never thought that the relationships that another IRB member had with industry affected his or her IRB-related decisions in an inappropriate way, 11.9% said they thought this occurred rarely, 2.4% thought it occurred sometimes, and 0.2% thought it occurred often. Seventy-eight respondents (15.1%) reported that at least one protocol came before their IRB during the previous year that was sponsored either by a company with which they had a relationship or by a competitor of that company, both of which could be considered conflicts of interest. Of these 78 members (62 voting members and 16 nonvoting members), 57.7% reported that they always disclosed the relationship to an IRB official, 7.7% said they sometimes did, 11.5% said they rarely did, and 23.1% said they never did. Of the 62 voting members who reported conflicts, 64.5% reported that they never voted on the protocol, 4.8% said they rarely did, 11.3% said they sometimes did, and 19.4% said they always did. Most respondents reported that the views of IRB members who had experience working with industry were beneficial in reviewing industry-sponsored protocols. CONCLUSIONS: Relationships between IRB members and industry are common, and members sometimes participate in decisions about protocols sponsored by companies with which they have a financial relationship. Current regulations and policies should be examined to be sure that there is an appropriate way to handle conflicts of interest stemming from relationships with industry. PMID- 17135586 TI - Patients' views on financial conflicts of interest in cancer research trials. AB - BACKGROUND: Financial ties between researchers or medical centers and companies whose drugs are being tested have come under increasing scrutiny. METHODS: We conducted in-person interviews with 253 patients in cancer-research trials (a 93% response rate) at five U.S. medical centers to determine their attitudes regarding potential financial conflicts of interest among researchers and medical centers. RESULTS: More than 90% of patients expressed little or no worry about financial ties that researchers or institutions might have with drug companies. Most patients said they would have enrolled in the trial even if the drug company had paid the researcher for speaking (82% of those interviewed) or consulting (75%) or if the researcher had received royalty payments (70%) or owned stock in the company (76%). Similarly, most patients would have enrolled in the trial if their cancer center had owned stock in the drug company (77%) or received royalty payments from the company (79%). Most patients believed it was ethical for researchers to receive speaking fees (81%) or consulting fees (82%) from the company. However, a substantial minority of patients wanted disclosure of the oversight system for researchers (40%) and of researchers' financial interests (31%); 17% thought no disclosure to patients was necessary. CONCLUSIONS: Most patients in cancer-research trials were not worried about financial ties between researchers or medical centers and drug companies and would still have enrolled in the trial if they had known about such financial ties. A substantial minority wanted to be informed about the oversight system to protect against financial conflicts of interest and about researchers' financial interests. PMID- 17135587 TI - Clinical practice. Management of menopausal symptoms. PMID- 17135588 TI - Images in clinical medicine. Bisphosphonate-associated osteonecrosis of the jaw. PMID- 17135589 TI - Case records of the Massachusetts General Hospital. Case 37-2006. A 19-year-old woman with thyroid cancer and lower gastrointestinal bleeding. PMID- 17135590 TI - Getting smarter--the toxicity of undertreated HIV infection. PMID- 17135591 TI - Thiazolidinediones for nonalcoholic steatohepatitis--promising but not ready for prime time. PMID- 17135592 TI - Conflict of interest--or conflict of priorities? PMID- 17135593 TI - Statin therapy after stroke or transient ischemic attack. PMID- 17135594 TI - Risks and benefits of celecoxib to prevent colorectal adenomas. PMID- 17135595 TI - Neonatal MRI and neurodevelopmental outcomes. PMID- 17135596 TI - Case 21-2006: a man with left-sided facial pain. PMID- 17135597 TI - Tracking neural stem cells in patients with brain trauma. PMID- 17135598 TI - No evidence for a major role of polymorphisms during bupropion treatment. AB - This study evaluated the ability of polymorphisms in five candidate genes to predict weight gain among patients taking bupropion or placebo in a smoking cessation trial. Five hundred fifty-three smokers were enrolled into a randomized double-blind, placebo-controlled trial and followed for 12 months. Five candidate genes [DRD2 Taq1 (rs1800497), DRD2-141 (rs1799732), C957T (rs6277), COMT (rs4818), and SLC6A3] were genotyped. Weights at baseline, at end of treatment, and after 6 and 12 months of follow-up were self-reported. Smoking abstinence at each endpoint was self-reported and confirmed biochemically. A self-reported average weight gain after 12 months of 1.1 +/- 6.0 kg (mean +/- standard deviation) in the bupropion group and 1.8 +/- 4.8 kg in the placebo group was noted. For subjects with biochemically confirmed abstinence from smoking, the HL genotype (alleles coding Val at codon 108 are denoted as H, and those coding Met are denoted as L) at the COMT locus and A1A1 genotype at the DRD2 Taq1 locus were associated with less weight gain at the end of treatment. The TC genotype at the C957T locus was associated with increased weight gain at 6 months of follow-up. However, no polymorphisms or their interactions with bupropion consistently and significantly predicted baseline BMI or weight change during treatment for all study subjects. Overall, our results do not support a major role for these five candidate genes in weight gain after smoking cessation. PMID- 17135599 TI - Interleukin-6 gene polymorphism -174G/C influences plasma lipid levels in women. AB - Elevated levels of the pro-inflammatory cytokine interleukin-6 (IL-6) have been associated with cardiovascular risk factors. The objective of this study was to investigate potential associations between the promoter polymorphism IL-6 -174G/C and the following indices of metabolism: BMI, waist-to-hip ratio, and plasma levels of IL-6, cholesterol, low-density lipoprotein, triglycerides, high-density lipoprotein, leptin, and C-reactive protein in 252 42-year-old women and 245 51 year-old men. Subgroups were also studied 5 years later. The CC genotype of the IL-6 polymorphism was associated with lower levels of cholesterol and low-density lipoprotein (p < 0.001) in women. This finding was replicated in the follow-up, when a significant association between the CC genotype and low triglycerides was also observed. The association between the C allele and lipid pattern found in women was not found in men, where on the contrary, C carriers tended to display elevated triglycerides. IL-6 genotype was not associated with IL-6 plasma levels in either sample. The results suggest different effects of the IL-6 polymorphism on metabolic indices in women and men. None of the associations between IL-6 genotype and lipid pattern seemed to result from an effect of the polymorphism on IL-6 plasma levels. PMID- 17135600 TI - ABCA1 single nucleotide polymorphisms on high-density lipoprotein-cholesterol and overweight: the D.E.S.I.R. study. AB - The adenosine triphosphate-binding cassette A1 (ABCA1) gene plays a key role in reverse cholesterol transport. Some ABCA1 gene polymorphisms have been associated with high-density lipoprotein-cholesterol (HDL-C) concentrations. The aim of this study was to assess the effect of three polymorphisms, C69T, G378C, and G1051A (R219K), on HDL-C levels and their interaction with BMI in more than 5000 French whites from the D.E.S.I.R. (Data from an Epidemiological Study on the Insulin Resistance syndrome) cohort study. The T allele of the C69T single nucleotide polymorphism (SNP) was associated with higher HDL-C levels in normal-weight men (BMI <25 kg/m(2)). The C allele of the G378C SNP was associated with lower HDL-C in overweight subjects (BMI > or =25 kg/m(2)). For the G1051A SNP, in the normal weight group, the minor A allele was significantly associated with higher HDL-C levels. In contrast, in overweight people, the minor allele was associated with lower HDL-C levels. After accounting for multiple testing, empiric p values remained significant for the associations between G378C SNP and HDL-C in the overweight group and between G1051A SNP and HDL-C in the normal-weight group. This study suggests that ABCA1 gene polymorphisms modulate HDL-C concentrations, in interaction with BMI, and, thus, they might influence cardiovascular risk in the general population. PMID- 17135601 TI - Fetal and maternal peroxisome proliferator-activated receptor gamma2 Pro12Ala does not influence birth weight. AB - The association between the peroxisome proliferator-activated receptor (PPAR)gamma2 Pro12Ala polymorphism and insulin resistance is reported to depend on low birth weight. Low birth weight itself has been linked to type 2 diabetes and cardiovascular diseases in adulthood. We assessed whether the PPARgamma2 Pro12Ala polymorphism determines body size at birth and whether metabolic differences between the genotypes are already detectable in the newborn. This study was conducted at the obstetrics department of the Charite, Berlin, Germany. One thousand nine hundred thirty white woman/child pairs were consecutively included and genotyped. The newborn's weight, length, and head circumference were measured. Total glycated hemoglobin in blood served as a surrogate of fetal insulin resistance and glucose use. We found that neither the fetal nor the maternal Pro12Ala genotype determined body size or total glycated hemoglobin at birth. The results suggest that the PPARgamma2 Pro12Ala polymorphism is not relevant for intrauterine growth. Previously reported effects of PPARgamma2 Pro12Ala on insulin resistance seem to arise later in life. PMID- 17135602 TI - Increase in visfatin after weight loss induced by gastroplastic surgery. AB - OBJECTIVE: The recently described adipokine visfatin is produced in visceral fat and has been suggested to influence insulin resistance. To investigate whether visfatin concentrations are related to changes in body weight, this adipokine was measured in insulin-resistant severely obese patients before and after gastroplastic surgery. RESEARCH METHODS AND PROCEDURES: Visfatin, interleukin-6, high-sensitivity C-reactive protein, homeostasis model assessment of insulin resistance (HOMA-IR), and other clinical parameters were assessed in 36 severely obese subjects (28 female; mean age, 43 years) with a median BMI of 44.3 kg/m(2) (95% confidence interval, 43.3 to 48.1 kg/m(2)). RESULTS: After surgery, BMI decreased to a median of 31.9 kg/m(2) (30.1 to 35.1 kg/m(2)) (p < 0.0001). Median visfatin concentrations increased significantly after weight loss [70.9 ng/mL (61.4 to 75.6 ng/mL) vs. 86.4 ng/mL (79.4 to 89.8 ng/mL); p < 0.0005]. This increase correlated with the decrease of insulin and HOMA-IR and was associated with a reduction in plasma interleukin-6 and high-sensitivity C-reactive protein concentrations. DISCUSSION: Massive weight loss after gastroplastic surgery is accompanied by an increase in circulating concentrations of the novel adipokine visfatin. This increase correlates with the decrease in plasma insulin concentrations and HOMA-IR. PMID- 17135603 TI - Regulation of secreted protein acidic and rich in cysteine during adipose conversion and adipose tissue hyperplasia. AB - OBJECTIVE: To explore the regulation of secreted protein acidic and rich in cysteine (SPARC) expression and its role in adipose tissue. RESEARCH METHODS AND PROCEDURES: We studied the regulation of SPARC expression in transgenic mice expressing the human beta3 and alpha2 adrenergic receptors on a murine beta3 adrenergic receptor null background that became obese under a high-fat diet mainly as a result of adipose tissue hyperplasia. Furthermore, we analyzed its expression in human adipose tissue and its regulation during adipocyte differentiation. RESULTS: SPARC protein in adipose tissue was increased in obese transgenic mice compared with control mice, indicating that SPARC expression was associated with adipose tissue hyperplasia. Both SPARC mRNA and protein were detected in human adipose tissue. Comparing adipocytes and vascular stroma, we found that SPARC expression was mainly associated with the adipocyte fraction. Consistent with this, SPARC transcript increased during differentiation of human primary preadipocytes. 3T3-L1 preadipocytes showed an increase in SPARC expression in differentiated cells but with biphasic expression during the process. After induction in committed cells, SPARC mRNA and protein levels declined as differentiation began and returned to elevated levels in fully differentiated adipocytes. DISCUSSION: SPARC expression correlated with adipose tissue hyperplasia and adipogenesis. Therefore, SPARC seems to play a role in adipose tissue physiology as it is involved in growth and differentiation. PMID- 17135604 TI - Differential regulation of adiponectin receptor gene expression by adiponectin and leptin in myotubes derived from obese and diabetic individuals. AB - OBJECTIVE: This study aimed to investigate the regulation of adiponectin receptors 1 (AdipoR1) and 2 (AdipoR2) gene expression in primary skeletal muscle myotubes, derived from human donors, after exposure to globular adiponectin (gAd) and leptin. RESEARCH METHODS AND PROCEDURES: Four distinct primary cell culture groups were established [Lean, Obese, Diabetic, Weight Loss (Wt Loss); n = 7 in each] from rectus abdominus muscle biopsies obtained from surgical patients. Differentiated myotube cultures were exposed to gAd (0.1 microg/mL) or leptin (2.5 microg/mL) for 6 hours. AdipoR1 and AdipoR2 gene expression was measured by real-time polymerase chain reaction analysis. RESULTS: AdipoR1 mRNA expression in skeletal muscle myotubes derived from Lean subjects (p < 0.05) was stimulated 1.8 fold and 2.5-fold with gAd and leptin, respectively. No increase in AdipoR1 gene expression was measured in myotubes derived from Obese, Diabetic, or Wt Loss subjects. AdipoR2 mRNA expression was unaltered after gAd and leptin exposure in all myotube groups. DISCUSSION: Adiponectin and leptin are rapid and potent stimulators of AdipoR1 in myotubes derived from lean healthy individuals. This effect was abolished in myotubes derived from obese, obese diabetic subjects, and obese-prone individuals who had lost significant weight after bariatric surgery. The incapacity of skeletal muscle of obese and diabetic individuals to respond to exogenous adiponectin and leptin may be further suppressed as a result of impaired regulation of the AdipoR1 gene. PMID- 17135605 TI - Human adenovirus 36 induces adiposity, increases insulin sensitivity, and alters hypothalamic monoamines in rats. AB - OBJECTIVE: Human adenovirus 36 (Ad-36) increases adiposity and reduces serum lipids in chicken, mouse, and non-human primate models, and it is linked to obesity in sero-epidemiological studies in humans. Involvement of the central nervous system (CNS) or adipose tissue in the mechanism of Ad-36-induced adiposity is unknown. The effects of Ad-36 on adiposity and on the neuroendocrine system were investigated in a rat model. RESEARCH METHODS AND PROCEDURES: Five week-old male Wistar rats were inoculated intraperitoneally with Ad-36 or medium. RESULTS: Despite similar food intakes, infected rats attained significantly greater body weight and fat pad weight by 30 weeks post-inoculation. Epididymal inguinal, retroperitoneal, and visceral fat pad weights of the infected group were greater by 60%, 46%, and 86%, respectively (p < 0.00001). The fasting serum insulin level and homeostasis model assessment index indicated greater insulin sensitivity in the infected group. Visceral adipose tissue expression of glycerol 3-phosphate dehydrogenase, peroxisome proliferator-activated receptor gamma, and CCAAT/enhancer-binding protein alpha and beta was markedly increased in the infected animals compared with controls. Ad-36 decreased norepinephrine levels significantly in the paraventricular nucleus in infected vs. control rats (mean +/- standard error, 8.9 +/- 1.1 vs. 12.8 +/- 1.2 pg/microg protein; p < 0.05). Ad 36 markedly decreased serum corticosterone in infected vs. control rats (mean +/- standard error, 97 +/- 41.0 vs. 221 +/- 111 ng/mL; p < 0.005). DISCUSSION: The results suggest that the pro-adipogenic effect of Ad-36 may involve peripheral as well as central effects. The male Wistar rat is a good model for the elucidation of metabolic and molecular mechanisms of Ad-36-induced adiposity. PMID- 17135606 TI - Heat production and body temperature during cooling and rewarming in overweight and lean men. AB - OBJECTIVE: To compare overweight and lean subjects with respect to thermogenesis and physiological insulation in response to mild cold and rewarming. RESEARCH METHODS AND PROCEDURES: Ten overweight men (mean BMI, 29.2 +/- 2.8 kg/m(2)) and 10 lean men (mean BMI, 21.1 +/- 2.0 kg/m(2)) were exposed to cold air for 1 hour, followed by 1 hour of rewarming. Body composition was determined by hydrodensitometry and deuterium dilution. Heat production and body temperatures were measured continuously by indirect calorimetry and thermistors, respectively. Muscle activity was recorded using electromyography. RESULTS: In both groups, heat production increased significantly during cooling (lean, p = 0.004; overweight, p = 0.006). The increase was larger in the lean group compared with the overweight group (p = 0.04). During rewarming, heat production returned to baseline in the overweight group and stayed higher compared with baseline in the lean group (p = 0.003). The difference in heat production between rewarming and baseline was larger in the lean (p = 0.01) than in the overweight subjects. Weighted body temperature of both groups decreased during cold exposure (lean, p = 0.002; overweight, p < 0.001) and did not return to baseline during rewarming. DISCUSSION: Overweight subjects showed a blunted mild cold-induced thermogenesis. The insulative cold response was not different among the groups. The energy efficient response of the overweight subjects can have consequences for energy balance in the long term. The results support the concept of a dynamic heat regulation model instead of temperature regulation around a fixed set point. PMID- 17135607 TI - Resistance training lowers exercise-induced oxidative stress and homocysteine levels in overweight and obese older adults. AB - OBJECTIVE: To compare exercise-induced oxidative stress and levels of homocysteine and cholesterol in normal-weight and overweight older adults after resistance exercise (RX). RESEARCH METHODS AND PROCEDURES: This interventional study was conducted at a wellness center. Forty-nine older adults (age range, 60 to 72 years) were stratified by BMI (<25 kg/m(2) normal weight, > or =25 kg/m(2) overweight/obese) and then randomly assigned to either a control non-exercise group or an RX group. The RX group completed a 6-month training program. Exercise induced lipid hydroperoxides (PEROXs) and thiobarbituric-reactive acid substances, homocysteine, lipoprotein a, cholesterol, and high-density lipoprotein cholesterol were measured before and after the 6-month RX program. RESULTS: PEROXs and thiobarbituric-reactive acid substances were lower in both the overweight/obese and normal-weight RX-trained groups compared with control groups (p < 0.05). Homocysteine levels were lower in both overweight/obese and normal-weight RX groups compared with control groups (p < 0.05). Lipoprotein a, total cholesterol, and high-density lipoprotein cholesterol were not different in normal-weight and overweight/obese groups before or after RX. The change in muscle strength was correlated with homocysteine at 6 months (r = -0.452, p < 0.05), whereas the change in PEROXs was correlated with the change in body fat (r = -0.329). DISCUSSION: To our knowledge, these data are the first to show that RX reduces exercise-induced oxidative stress and homocysteine regardless of adiposity, indicating that this protection can be afforded in an older, overweight/obese population as effectively as in healthy older adults. These data suggest that RX may afford some protection against emerging cardiovascular risk factors using a mode of exercise that supports body weight. PMID- 17135608 TI - Serum phospholipid fatty acids, adipose tissue, and metabolic markers in obese adolescents. AB - OBJECTIVE: Fatty acid (FA) composition has a role in adipogenesis. The objective was to study serum phospholipid (PL) FAs in adolescents and their relation to abdominal adipose tissue (AT) compartments and metabolic markers. RESEARCH METHODS AND PROCEDURES: Abdominal AT was measured by magnetic resonance imaging and FA pattern was determined in serum PL of 10 obese adolescents (5 females), median age 12.0 years (range, 10.4 to 16.4) and BMI 30.7 (26.8 to 40.4), and 15 lean control subjects (9 females), median age 12.6 years (range, 11.3 to 15.4), and BMI 19.5 (17.1 to 23.4). RESULTS: Obese adolescents had relatively higher levels of saturated FA (SFA) and nervonic acid compared with controls. Serum PL concentration of n-3 polyunsaturated fatty acids (PUFA) was lower in the obese vs. lean females (p = 0.01), including docosahexaenoic acid (DHA) (p = 0.01). The ratios of arachidonic acid to DHA and total n-6/n-3 FA were increased in obese children (p = 0.02 and 0.01, respectively). n-3 PUFAs were inversely correlated to all subcutaneous AT compartments except visceral AT. The homeostasis model assessment index of beta-cell function related inversely to DHA concentration (p = 0.03). All changes were more marked in the females. DISCUSSION: Serum FA pattern in obese adolescents differed significantly from that in age-matched lean controls, reflecting a decrease in n-3 PUFA, especially DHA, and an increase in SFA. The subcutaneous AT, but not visceral AT, correlated to the changes in PUFA and SFA, suggesting an abnormal essential FA metabolism in obese adolescents. PMID- 17135609 TI - True fractional calcium absorption is decreased after Roux-en-Y gastric bypass surgery. AB - OBJECTIVE: Roux-en-Y gastric bypass (RYGB) is considered to be the gold standard alternative treatment for severe obesity. Weight loss after RYGB results primarily from decreased food intake. Inadequate calcium (Ca) intake and metabolic bone disease can occur after gastric bypass. To our knowledge, whether malabsorption of Ca contributes to an altered Ca metabolism in the RYGB patient has not been addressed previously. RESEARCH METHODS AND PROCEDURES: We recruited 25 extremely obese women in order to study true fractional Ca absorption (TFCA) before and 6 months after RYGB surgery, using a dual stable isotope method ((42)Ca and (43)Ca) and test load of Ca (200 mg). Hormones regulating Ca absorption and markers of bone turnover were also measured. RESULTS: In 21 women (BMI 52.7 +/- 8.3 kg/m(2), age 43.9 +/- 10.4 years) who successfully completed the study, TFCA decreased from 0.36 +/- 0.08 to 0.24 +/- 0.09 (p < 0.001) after RYGB. Bone turnover markers increased significantly (p < 0.01). TFCA correlated with estradiol levels (r = 0.512, p < 0.02) and tended to correlate with 1,25 (OH)(2)D (r = 0.427, p < 0.06) at final measurement. Stepwise linear regression indicated that estradiol explained 62% of the variance for TFCA at 6 months post surgery (p < 0.01). DISCUSSION: TFCA decreases (0.12 +/- 0.08) after RYGB surgery but remains within normal range. Although only some patients were estimated to have low Ca absorption after surgery, all of the patients showed a dramatic increase in markers of bone resorption. The alteration in Ca metabolism after RYGB-induced weight loss appears to be regulated primarily by estradiol levels and might ultimately affect bone mass. PMID- 17135610 TI - The feet of overweight and obese young children: are they flat or fat? AB - OBJECTIVE: The purpose of this study was to determine whether the flat feet displayed by young obese and overweight children are attributable to the presence of a thicker midfoot plantar fat pad or a lowering of the longitudinal arch relative to that in non-overweight children. RESEARCH METHODS AND PROCEDURES: Foot anthropometry, an arch index derived from plantar footprints, and midfoot plantar fat pad thickness measured by ultrasound were obtained for 19 overweight/obese preschool children (mean age, 4.3 +/- 0.9 years; mean height, 1.07 +/- 0.1 m; mean BMI, 18.6 +/- 1.2 kg/m(2)) and 19 non-overweight children matched for age, height, and sex (mean age, 4.3 +/- 0.7 years; mean height, 1.05 +/- 0.1 m; mean BMI, 15.7 +/- 0.7 kg/m(2)). RESULTS: Independent t tests revealed no significant between-subject group differences (p = 0.39) in the thickness of the midfoot plantar fat pad. However, the overweight/obese children had a significantly lower plantar arch height (0.9 +/- 0.3 cm) than their non overweight counterparts (1.1 +/- 0.2 cm; p = 0.04). DISCUSSION: The lower plantar arch height found in the overweight/obese children suggests that the flatter feet characteristic of overweight/obese preschool children may be caused by structural changes in their foot anatomy. It is postulated that these structural changes, which may adversely affect the functional capacity of the medial longitudinal arch, might be exacerbated if excess weight bearing continues throughout childhood and into adulthood. PMID- 17135611 TI - Inhibitory effect of obesity on gonadotropin, estradiol, and inhibin B levels in fertile women. AB - OBJECTIVE: To examine whether obesity and insulin resistance have an independent effect on the gonadotropin, estradiol, and inhibin B serum levels and follicle count in the early follicular phase of fertile women with a wide range of BMI and without signs of hyperandrogenism. RESEARCH METHODS AND PROCEDURES: Twenty-two overweight and obese (BMI > or =25.0 kg/m(2)) women and 10 normal-weight (BMI <25.0 kg/m(2)) women, all having apparently normal fertility, were studied. Serum concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol, inhibin B, and insulin, level of insulin resistance (estimated by homeostasis model assessment for insulin resistance), and follicle count were measured during the early follicular phase (Days 2 to 5 of the menstrual cycle). RESULTS: Overweight women showed lower FSH (p < 0.001), LH (p < 0.001), and inhibin B (p < 0.05) levels compared with normal-weight women, whereas estradiol concentrations and follicle count were not significantly different between the two groups. When normal-weight and overweight women were examined as a group and multiple regression analyses were performed, estradiol showed a negative association with BMI (or waist circumference) (p < 0.05) and a positive correlation with LH (p < 0.05) and FSH (p < 0.05); inhibin B maintained a positive association only with estradiol (p < 0.05); and FSH and LH showed a negative correlation with BMI (or waist circumference) (p < 0.001 and p < 0.01, respectively). DISCUSSION: Overweight and obese fertile women have lower FSH, LH, inhibin B, and estradiol levels in the early follicular phase, with a possible direct inhibitory effect of body mass on gonadotropin and estradiol production, independently of age, insulin (concentrations and sensitivity), and other hormones. By contrast, the number of ovary follicles does not seem to be influenced by insulin and body mass in these patients. PMID- 17135612 TI - A simple venous thromboembolism prophylaxis protocol for patients undergoing bariatric surgery. AB - OBJECTIVE: Pulmonary embolism is a leading cause of death for bariatric patients. Numerous regimens have been proposed, but a comprehensive, simple approach is lacking. This study provides a simple, easily implemented prophylaxis regimen. RESEARCH METHODS AND PROCEDURES: One hundred fifty bariatric surgery patients were evaluated. Patients considered at high risk for venous thromboembolism had heart failure, a BMI of >/=50 kg/m(2), or a history of venous thromboembolism or pelvic surgery. Preoperatively and postoperatively, all patients received subcutaneous enoxaparin or unfractionated heparin. High-risk patients received either preoperatively inserted inferior vena cava filters or continuous heparin infusions intraoperatively. All high-risk patients were anticoagulated with warfarin (Coumadin; Bristol Myers-Squibb, Princeton, NJ) for at least 3 months postoperatively. Initially, some patients experienced significant hemorrhage; to prevent this, sutures were oversewn into staple lines. RESULTS: No patient experienced venous thromboembolism; a binomial test showed that the regimen reduced the risk of this complication to less than 2% (p < 0.05). Hemorrhage sufficient to require transfusion occurred in 4 of the first 20 patients; of the remaining 130 patients, into whose staple lines sutures were oversewn, none required transfusion (p < 0.05). DISCUSSION: Patients should be divided into those who are at high risk and those who are at low risk for venous thromboembolism. All patients should receive pre- and postoperative anticoagulation. High-risk patients should also receive either an inferior vena cava filter or intraoperative heparin infusions, as well as at least 3 months of Coumadin therapy. Oversewing of staple lines may reduce the risk of hemorrhage. PMID- 17135613 TI - Soda consumption and overweight status of 2-year-old mexican-american children in california. AB - OBJECTIVE: The prevalence of overweight in United States children, 2 to 5 years old, has increased 2-fold since 1975, with the highest prevalence in Mexican Americans. The objective of this study was to determine the association between current soda consumption and overweight in 2-year-old Mexican-American children. RESEARCH METHODS AND PROCEDURES: The Center for the Health Assessment of Mothers and Children of Salinas study is a longitudinal study of the health of low-income Latino pregnant women and their children living in the Salinas Valley, CA. Six hundred pregnant women were enrolled (October 1999 to October 2000), and their children were followed until 2 years of age. This cross-sectional analysis includes the 354 children who completed the 2-year follow-up interview. Standing height (centimeters) and weight (grams) were measured at 2 years. Overweight was defined as > or =95th percentile of the sex-specific BMI for each child's age. RESULTS: Fifty-five (15.5%) children were overweight. Over half (56%) reported consuming any soda in the last week. After covariate adjustment, compared with no soda consumption, <1 soda/d was not related to overweight (adjusted odds ratio, 0.97; 95% confidence interval, 0.47, 1.99), but > or =1 soda/d was significantly associated with overweight (adjusted odds ratio, 3.39; 95% confidence interval, 1.43, 8.07), and the test for trend was significant (p = 0.02). DISCUSSION: At 2 years of age, the prevalence of overweight among the Center for the Health Assessment of Mothers and Children of Salinas cohort is higher than the national prevalence estimate for Mexican-American 2- to 5-year-old children and is significantly associated with current soda consumption. Interventions to reduce consumption of soda in young Mexican-American children should be considered. PMID- 17135614 TI - Dose-dependent effects of training and detraining on weight in 6406 runners during 7.4 years. AB - OBJECTIVE: Prior randomized and non-randomized training studies have failed to establish a dose-response relationship between vigorous exercise and weight loss; this failure may be due, in part, to their short durations and small sample sizes. The objectives of this study were to determine whether exercise reduces body weight and to examine the dose-response relationships between changes in exercise and changes in total and regional adiposity. RESEARCH METHODS AND PROCEDURES: This was a large prospective study of 3973 men and 1444 women who quit running (detraining), 270 men and 146 women who started running (training), and 420 men and 153 women who remained sedentary during 7.4 years of follow-up. The outcomes measured were weekly running distance, body weight, BMI, body circumferences, and bra cup size. RESULTS: There were significant inverse relationships between the changes in the amount of vigorous exercise (km/wk run) and the changes in weight and BMI in men (slope +/- standard error: -0.039 +/- 0.005 kg/km per week and -0.012 +/- 0.002 kg/m(2) per km/wk, respectively) and in older women (-0.060 +/- 0.018 kg/km per week and -0.022 +/- 0.007 kg/m(2) per km/wk) who quit running, and in initially sedentary men (-0.098 +/- 0.017 kg/km per week and -0.032 +/- 0.005 kg/m(2) per km/wk) and women (-0.062 +/- 0.023 kg/km per week and -0.021 +/- 0.008 kg/m(2) per km/wk) who started running. Changes in waist circumference, an indicator of intra-abdominal fat, were also inversely related to changes in running distance in men who quit (-0.026 +/- 0.005 cm/km per week) or started running (-0.078 +/- 0.017 cm/km per week). DISCUSSION: The initiation of vigorous exercise and its cessation decrease and increase, respectively, body weight and intra-abdominal fat, and these changes are proportional to the change in exercise dose. PMID- 17135615 TI - Aerobic exercise and snoring in overweight children: a randomized controlled trial. AB - OBJECTIVE: To determine whether regular aerobic exercise improves symptoms of sleep-disordered breathing in overweight children, as has been shown in adults. RESEARCH METHODS AND PROCEDURES: Healthy but overweight (BMI > or =85th percentile) 7- to 11-year-old children were recruited from public schools for a randomized controlled trial of exercise effects on diabetes risk. One hundred children (53% black, 41% male) were randomly assigned to a control group (n = 27), a low-dose exercise group (n = 36), or a high-dose exercise group (n = 37). Exercise groups underwent a 13 +/- 1.5 week after-school program that provided 20 or 40 minutes per day of aerobic exercise (average heart rate = 164 beats per minute). Group changes were compared on BMI z-score and four Pediatric Sleep Questionnaire scales: Snoring, Sleepiness, Behavior, and a summary scale, Sleep Related Breathing Disorders. Analyses were adjusted for age. RESULTS: Both the high-dose and low-dose exercise groups improved more than the control group on the Snoring scale. The high-dose exercise group improved more than the low-dose exercise and control groups on the summary scale. No group differences were found for changes on Sleepiness, Behavior, or BMI z-score. At baseline, 25% screened positive for sleep-disordered breathing; half improved to a negative screen after intervention. DISCUSSION: Regular vigorous exercise can improve snoring, a symptom of sleep-disordered breathing, in overweight children. Aerobic exercise programs may be valuable for prevention and treatment of sleep-disordered breathing in overweight children. PMID- 17135616 TI - Categorization of low cardiorespiratory fitness using obesity indices in non smoking Singaporean women. AB - OBJECTIVE: To revisit cut-off values of BMI, waist circumference (WC), and waist to-stature ratio (WSR) based on their association with cardiorespiratory fitness (CRF). The derived cut-off points were compared with current values (BMI, 25.0 kg/m(2); WC, 80 cm) as recommended by the World Health Organization. RESEARCH METHODS AND PROCEDURES: Anthropometric indices were measured in a cross sectional study of 358 Singaporean female employees of a large tertiary hospital (63% Singaporean Chinese, 28% Malays, and 9% Indians). CRF was determined by the 1 mile walk test. Receiver operating characteristic curves were constructed to determine cut-off points. RESULTS: The cut-off points for BMI, WC, and WSR were 23.6 kg/m(2), 75.3 cm, and 0.48, respectively. The areas under the curve of BMI, WC, and WSR were 0.68, 0.74, and 0.74, respectively. For a given BMI, women with low CRF had higher WSR compared with women with high CRF. DISCUSSION: These findings provide convergent evidence that the cut-off points for Singaporean women were lower than the World Health Organization's criteria but were in good agreement with those reported for Asians. PMID- 17135617 TI - Validation and calibration of an accelerometer in preschool children. AB - OBJECTIVE: Obesity rates in young children are increasing, and decreased physical activity is likely to be a major contributor to this trend. Studies of physical activity in young children are limited by the lack of valid and acceptable measures. The purpose of this study was to calibrate and validate the ActiGraph accelerometer for use with 3- to 5-year-old children. RESEARCH METHODS AND PROCEDURES: Thirty preschool children wore an ActiGraph accelerometer (ActiGraph, Fort Walton Beach, FL) and a Cosmed portable metabolic system (Cosmed, Rome, Italy) during a period of rest and while performing three structured physical activities in a laboratory setting. Expired respiratory gases were collected, and oxygen consumption was measured on a breath-by-breath basis. Accelerometer data were collected at 15-second intervals. For cross-validation, the same children wore the same instruments while participating in unstructured indoor and outdoor activities for 20 minutes each at their preschool. RESULTS: In calibrating the accelerometer, the correlation between Vo(2) (ml/kg per min) and counts was r = 0.82 across all activities. The only significant variable in the prediction equation was accelerometer counts (R(2) = 0.90, standard error of the estimate = 4.70). In the cross-validation, the intraclass correlation coefficient between measured and predicted Vo(2) was R = 0.57 and the Spearman correlation coefficient was R = 0.66 (p < 0.001). Cut-off points for moderate- and vigorous intensity physical activity were identified at 420 counts/15 s (Vo(2) = 20 mL/kg per min) and 842 counts/15 s (Vo(2) = 30 mL/kg per min), respectively. When these cutpoints were applied to the cross-validation data, percentage agreement, kappa, and modified kappa for moderate activity were 0.69, 0.36, and 0.38, respectively. For vigorous activity, the same measures were 0.81, 0.13, and 0.62. DISCUSSION: Accelerometer counts were highly correlated with Vo(2) in young children. Accelerometers can be appropriately used as a measure of physical activity in this population. PMID- 17135618 TI - A randomized clinical trial testing treatment preference and two dietary options in behavioral weight management: preliminary results of the impact of diet at 6 months--PREFER study. AB - OBJECTIVE: The PREFER study objectives were to examine potential differences in weight loss during a standard behavioral intervention between subjects assigned to one of two calorie- and fat-restricted diets [standard behavior treatment (SBT) and lacto-ovo-vegetarian ([SBT+LOV)], with or without regard to their preferred dietary treatment. This article reports the differences in outcomes between diet groups after the first 6 months of the intervention. RESEARCH METHODS AND PROCEDURES: The study used a four-group design. Subjects (n = 182) were randomized to a treatment preference group and then to a dietary treatment group. For this report, preference groups were combined to permit comparisons by dietary treatment only (SBT, n = 98; SBT+LOV, n = 84). Additional analyses compared SBT+LOV subjects who were 100% adherent (did not consume any meat, fish, or poultry, n = 47) to those who were <100% adherent (n = 24). RESULTS: Significant differences were seen in the baseline to 6-month change scores between the two groups for carbohydrate consumption (p = 0.013), protein consumption (p < 0.001), polyunsaturated-to-saturated fat ratio (p = 0.009), and low-density lipoprotein-cholesterol (LDL-C) level (p = 0.013). Among SBT+LOV subjects, those who were 100% adherent experienced greater reductions in weight (p < 0.001), total cholesterol (p = 0.026), LDL-C (p = 0.034), and glucose (p = 0.002) and consumed less fat (p = 0.030) compared with those who were <100% adherent. DISCUSSION: Differences between dietary treatment groups at 6 months were minimal, most likely because one-third of the SBT+LOV group did not follow the vegetarian diet and because both groups had the same calorie and fat restrictions. SBT+LOV subjects who were 100% adherent were more successful at both weight loss and cholesterol reduction than those who were <100% adherent, suggesting that vegetarian diets are efficacious for weight and cholesterol control. PMID- 17135619 TI - Increased resting energy expenditure after 40 minutes of aerobic but not resistance exercise. AB - OBJECTIVE: Resting energy expenditure (REE) is increased 24 hours after high intensity aerobic exercise lasting 60 minutes, whereas results have been inconsistent after resistance training and aerobic exercise of shorter duration. The objective of the study was to compare the effects of 40 minutes of high intensity aerobic vs. resistance exercise on REE 19 to 67 hours after exercise. RESEARCH METHODS AND PROCEDURES: REE was compared 19, 43, and 67 hours after 40 minutes of aerobic training (AT; 80% maximum heart rate) or resistance training (RT; 10 repetitions at 80% maximum strength, two sets and eight exercises). Twenty-three black and 22 white women were randomly assigned to AT, RT, or no training (controls). Exercisers trained 25 weeks. REE was measured after a 12 hour fast. RESULTS: There was a significant time x group interaction for REE when adjusted for fat-free mass and fat mass, with post hoc tests revealing that the 50-kcal difference between 19 and 43 hours (1310 +/- 196 to 1260 +/- 161 kcal) and the 34-kcal difference between 19 and 67 hours (1310 +/- 196 to 1276 +/- 168 kcal) were significant for AT. No other differences were found, including RT (19 hours, 1256 +/- 160; 43 hours, 1251 +/- 160; 67 hours, 1268 +/- 188 kcal). Urine norepinephrine increased with training only in AT. After adjusting for fat-free mass, REE Delta between 19 and both 43 and 67 hours was significantly related to urine norepinephrine (r = 0.76, p < 0.01 and 0.68, p < 0.03, respectively). DISCUSSION: Consistent with findings on longer duration AT, these results show that 40 minutes of AT elevates REE for 19 hours in trained black and white women. This elevation did not occur with 40 minutes of RT. Results suggest that differences are, in part, due to increased sympathetic tone. PMID- 17135620 TI - Maternal feeding strategies, child eating behaviors, and child BMI in low-income African-American preschoolers. AB - OBJECTIVE: To test the hypothesis that low-income African-American preschool children would have a higher BMI if their mothers reported greater "restriction" and "control" in feeding and if mothers reported that children showed greater "food responsiveness" and "desire to drink." In addition, to test whether higher maternal "pressure to eat" would be associated with lower child BMI. RESEARCH METHODS AND PROCEDURES: A questionnaire was completed by 296 low-income African American mothers of preschool children. It assessed three constructs on maternal feeding strategies ("restriction," "pressure to eat," and "control") and two on child eating behaviors ("food responsiveness" and "desire to drink"). Children's BMI was measured, and mothers' BMI was self-reported. RESULTS: The mean (standard deviation) BMI z-score of the children was 0.34 (1.5), and 44% of the mothers were obese (BMI > or =30 kg/m(2)). Only maternal "pressure to eat" had a significant overall association with child BMI z-score (r = -0.16, p < 0.01). Both maternal "restriction" and "control" were positively associated with children's BMI z-score in the case of obese mothers (r = 0.20, p = 0.03 and r = 0.24, p = 0.007, respectively), but this was not so in the case of non-obese mothers (r = -0.16, p = 0.05 and r = -0.07, p = 0.39, respectively). DISCUSSION: Among low-income African Americans, the positive association between maternal restriction and control in feeding and their preschoolers' BMI was limited to obese mothers. Relations between parent feeding strategies and child weight status in this population may differ on the basis of maternal weight status. PMID- 17135621 TI - The association of television and video viewing with fast food intake by preschool-age children. AB - OBJECTIVE: To examine the extent to which television (TV) and video viewing is associated with consumption of fast food by preschool-age children. RESEARCH METHODS AND PROCEDURES: In a cross-sectional study of 240 parents of children ages 2.0 to 5.9 years, parents reported the number of hours their child watched TV/videos on an average weekday and weekend day in the past month; a daily, weighted average of TV/video viewing was then calculated. The main outcome was parents' report of their children's fast food intake, using the question, "How many times a week does your child eat at fast food restaurants such as McDonald's, Burger King, or Kentucky Fried Chicken?" dichotomized to (never/<1 vs. > or =1 time/wk). The association of TV/video viewing with fast food intake was evaluated by multiple logistic regression before and after adjusting for several potential confounders. RESULTS: Twenty-two percent of parents reported that their child ate at fast food restaurants at least once per week. After adjusting for parents' age, race/ethnicity, and household income as well as child's age and sex, for each 1-hour increase of TV/video watched per day, the odds ratio (OR) for consuming fast food > or =1 time per week was 1.60 (95% confidence interval, 1.03 to 2.49). After further adjustment for socio environmental factors that might serve as proxies for the availability of healthy food options, such as parental time constraints and the availability and high cost of fresh fruits and vegetables in their neighborhoods, the OR for consuming fast food > or =1 time per week was minimally attenuated (OR, 1.55; 95% confidence interval, 1.04 to 2.31). DISCUSSION: TV/video viewing was correlated with fast food consumption among preschool children in this study. Our findings raise the possibility that greater exposure to TV and videos may influence preschool children's consumption of unhealthful foods. PMID- 17135622 TI - Health-related quality of life and weight loss among overweight and obese U.S. adults, 2001 to 2002. AB - OBJECTIVE: To examine the prevalence and association of health-related quality of life (HRQOL) with trying to lose weight and with weight loss practices (eating fewer calories, physical activity, and both) among overweight and obese U.S. adults >/= 20 years of age. RESEARCH METHODS AND PROCEDURES: This study used data from the 2001 to 2002 National Health and Nutrition Examination Survey, a continuous annual survey of the civilian non-institutionalized U.S. population. This analysis included those > or = 20 years of age with BMI > or = 25 (n = 2578) who responded to four standard HRQOL measures that assessed general health status and recent physical health, mental health, and activity limitation. RESULTS: Among obese men, but not women, there were significant increasing linear trends in the adjusted prevalence of trying to lose weight as physically unhealthy and activity limitation days increased. Regardless of BMI or HRQOL, reducing calories was a common weight loss practice (66% to 86%). Except for recent activity limitation, respondents with BMI > or = 35 did not generally differ by HRQOL level in the attainment of recommended physical activity either alone or in combination with reduced calories, whereas those in the BMI 25 to 34.9 groups often differed significantly by HRQOL level. Specifically, increased unhealthy or activity limitation days were associated with reduced prevalence of attained physical activity. DISCUSSION: Our findings indicate an association between trying to lose weight and a greater number of unhealthy days reported by obese men, suggesting that these men may be influenced by traditional clinical weight loss counseling that is prompted by weight and comorbidity, whereas women had a high prevalence of trying to lose weight irrespective of weight and HRQOL. Assessment of HRQOL, especially measures that evaluate physical domains, could provide subjective information to assist with weight counseling. PMID- 17135623 TI - Underreporting of BMI in adults and its effect on obesity prevalence estimations in the period 1998 to 2001. AB - OBJECTIVE: To identify the determinants of underreporting BMI and to evaluate the possibilities of using self-reported data for valid obesity prevalence rate estimations. RESEARCH METHODS AND PROCEDURES: A cross-sectional monitoring health survey was carried out between 1998 and 2002, and a review of published studies was performed. A total of 1809 men and 1882 women ages 20 to 59 years from The Netherlands were included. Body weight and height were reported and measured. Equations were calculated to estimate individuals' BMI from reported data. These equations and equations from published studies were applied to the present data to evaluate whether using these equations led to valid estimations of the obesity prevalence rate. Also, size of underestimation of obesity prevalence rate was compared between studies. RESULTS: The prevalence of obesity was underestimated by 26.1% and 30.0% among men and women, respectively, when based on reported data. The most important determinant of underreporting BMI was a high BMI. When equations to calculate individuals' BMI from reported data were used, the obesity prevalence rate was still underestimated by 12.9% and 8.1% of the "true" obesity prevalence rate among men and women, respectively. The degree of underestimating the obesity prevalence was inconsistent across studies. Applying equations from published studies to the present data led to estimations of the obesity prevalence varying from a 7% overestimation to a 74% underestimation. DISCUSSION: Valuable efforts for monitoring and evaluating prevention and treatment studies require direct measurements of body weight and height. PMID- 17135624 TI - Comparison of three bioelectrical impedance methods with DXA in overweight and obese men. AB - OBJECTIVE: To compare bioelectrical impedance analysis (BIA) of body composition using three different methods against DXA in overweight and obese men. RESEARCH METHODS AND PROCEDURES: Forty-three healthy overweight or obese men (ages 25 to 60 years; BMI, 28 to 43 kg/m(2)) underwent BIA assessment of body composition using the ImpediMed SFB7 (version 6; ImpediMed, Ltd., Eight Mile Plains, Queensland, Australia) in multifrequency mode (Imp-MF) and DF50 single-frequency mode (Imp-SF) and the Tanita UltimateScale (Tanita Corp., Tokyo, Japan). Validity was assessed by comparison against DXA using linear regression and limits of agreement analysis. RESULTS: All three BIA methods showed good relative agreement with DXA [Imp-MF: fat mass (FM), r(2) = 0.81; fat-free mass (FFM), r(2) = 0.81; percentage body fat (BF%), r(2) = 0.69; Imp-SF: FM, r(2) = 0.65; FFM, r(2) = 0.76; BF%, r(2) = 0.40; Tanita: BF%, r(2) = 0.44; all p < 0.001]. Absolute agreement between DXA and Imp-MF was poor, as indicated by a large bias and wide limits of agreement (bias, +/-1.96 standard deviation; FM, -6.6 +/- 7.7 kg; FFM, 8.0 +/- 7.1 kg; BF%, -7.0 +/- 6.6%). Imp-SF and Tanita exhibited a smaller bias but wide limits of agreement (Imp-SF: FM, -1.1 +/- 8.5 kg; FFM, 2.5 +/- 7.9 kg; BF%, -1.7 +/- 7.3%; Tanita: BF%, 1.2 +/- 9.5%). DISCUSSION: Compared with DXA, Imp-MF produced large bias and wide limits of agreement, and its accuracy estimating body composition in overweight or obese men was poor. Imp-SF and Tanita demonstrated little bias and may be useful for group comparisons, but their utility for assessment of body composition in individuals is limited. PMID- 17135625 TI - Correcting biases in estimates of mortality attributable to obesity. AB - OBJECTIVE: To assess whether a recent study that found a relatively small number of excess deaths attributable to obesity may have underestimated by not correcting for statistical biases. RESEARCH METHODS AND PROCEDURES: This prospective cohort study used data from the First National Health and Nutrition Examination Survey Epidemiologic Follow-Up Study. Survival analyses were conducted using 9690 individuals 32 to 87 years of age and 1886 all-cause deaths during a 9.1-year follow-up. Corrections were made for the reputed regression dilution bias by using the average BMI during the decade before follow-up as predictor. Corrections for the reputed reverse-causation bias were made by excluding participants with a history of serious illness. Attributable fractions were calculated and used to estimate excess deaths. RESULTS: The uncorrected estimate of excess deaths attributable to obesity (BMI > or =30) was 41.9, using 18.5 to 25 kg/m(2) as ideal-weight category. Using average BMI as predictor increased the estimate to 93.3. Correcting for reverse-causation effects increased the estimate further to 131.1 (range, 93.3 to 169.0). The uncorrected hazard ratio, 1.25, was increased to 1.41 by using average BMI as predictor, and then to 2.40 by correcting for reverse causation. Using BMI 21 to 25 kg/m(2) and 23 to 25 kg/m(2) as ideal-weight categories increased the corrected estimates to 144.6 (range, 80.5 to 177.2) and 164.1 (range, 103.8 to 194.9), respectively. Larger increases were found for overweight and Grade 2 to 4 obesity (BMI > or =35 kg/m(2)). For overweight, the uncorrected estimate using 18.5 to 25 kg/m(2) as ideal-weight category was -88.3 and the corrected estimate using 23 to 25 kg/m(2) as ideal-weight category was 205.4 (range, 114.5 to 296.3). DISCUSSION: Correcting for statistical biases and using higher ideal-weight categories increased the estimate of excess deaths attributable to obesity by approximately 400% and changed the negative estimate for overweight to a large positive estimate. PMID- 17135626 TI - Converging prevalences of obesity across educational groups in Switzerland. AB - OBJECTIVE: To assess whether the rapid increase in obesity prevalence among persons with higher education levels observed in one U.S. study is also observed in a European adult population. RESEARCH METHODS AND PROCEDURES: This study involved continuous surveillance of the adult population of Geneva, Switzerland (1993 to 2004), with annual random, independent, cross-sectional, representative samples (6635 men and 6558 women, ages 35 to 74 years) and analysis of 12-year trends in obesity prevalence across educational level subgroups. RESULTS: Obesity prevalence in men had an upward trend in the medium education subgroup (p < 0.02), a borderline upward trend in the high education subgroup (p < 0.08), but no trend in the low education subgroup. There was a borderline trend interaction between the male low and medium education subgroups (p < 0.09). Obesity prevalence in women had a borderline increase in the low education subgroup (p < 0.08), an almost borderline increase in the high education subgroup (p = 0.11), but no significant trend in the medium education subgroup. There was no evidence of trend interaction between the female education groups. DISCUSSION: In Geneva, as in the United States, the inverse association between education level and obesity rates has weakened over time among men, but, inconsistent with the U.S. findings, has persisted for women. Explanations may include more physically demanding occupations for men with low education levels and different attitudes toward body image between the sexes. PMID- 17135627 TI - Prevalence of metabolic syndrome and its relation to body composition in a Chinese rural population. AB - OBJECTIVE: To investigate the prevalence of metabolic syndrome (MetS) with three different working definitions in a rural Chinese population and to examine its relation to body composition. RESEARCH METHODS AND PROCEDURES: A total of 18,630 adults 25 to 64 years old (mean age, 45.8 years; 51.2% men) from 5686 families were enrolled from Anhui province of China during 2004 to 2005. Anthropometric measurement, body composition, blood pressure, plasma lipids, and fasting glucose and insulin and a questionnaire-based interview were obtained from each participant. Three different working definitions for MetS, including the U.S. National Cholesterol Education Program's Adult Treatment Panel III, a modified Adult Treatment Panel III that adopts the World Health Organization's criterion for central obesity in Asian populations, and one recently proposed by the International Diabetes Federation, were used in the study. RESULTS: According to the three definitions, the age-adjusted prevalence of MetS for adults 25 to 64 years old was 3.2%, 4.9%, and 3.9% in men and 7.2%, 11.5%, and 10.9% in women, respectively. MetS prevalence increases significantly with age in women, but not in men. Body fat percentage and BMI and waist circumference were significantly associated with each component of MetS, especially with triglyceride level, insulin resistance index, and number of MetS components (r = 0.28 to 0.49). DISCUSSION: The age-adjusted prevalence of MetS in our study population is lower than that reported in other urban Chinese populations. Significant gender differences in MetS prevalence were observed. The waist circumference is a good surrogate for abdominal fat percentage. PMID- 17135628 TI - Soluble intercellular adhesion molecules, soluble vascular cell adhesion molecules, and risk of coronary heart disease. AB - OBJECTIVE: We examined the association of circulating levels of soluble intercellular adhesion molecules (sICAM-1) and soluble vascular cell adhesion molecules (sVCAM-1) with coronary heart disease (CHD) risk factors and whether the adhesion molecules alone, and in combination, can serve as predictors of coronary CHD. RESEARCH METHODS AND PROCEDURES: Among 18,225 men from the Health Professional Follow-up Study who provided blood in 1994, we documented 266 incidents of non-fatal myocardial infarction or fatal CHD during 6 years of follow-up. The cases were matched 1:2 with non-cases on age, smoking, and month of blood draw. We found both adhesion molecules directly associated with BMI, inflammatory biomarkers, and triglycerides and inversely associated with high density lipoprotein and alcohol intake (p < 0.05). After adjustment for C reactive protein, cholesterol-to-high-density lipoprotein ratio, age, smoking, BMI, physical activity, alcohol intake, history of diabetes, parental history of CHD, aspirin use, antihypertensive drug use, and fasting status, the relative risk of CHD was 1.69 [95% confidence interval (CI), 1.14 to 2.51] for sICAM-1 and 1.34 (95% CI, 0.91 to 1.96) for sVCAM-1, when comparing the top quintile with the lower four quintiles. Control for other inflammatory or lipid biomarkers did not appreciably attenuate the associations. When we cross-classified participants based on their sICAM-1 and sVCAM-1 levels, only the men in the top quintile of both biomarkers [relative risk = 2.39 (95% CI, 1.45 to 3.91)] had a significantly elevated risk of CHD (P interaction = 0.01, multivariate model). DISCUSSION: sICAM-1 and sVCAM-1 are directly associated with obesity and other CHD risk factors. The combination of high levels of both adhesion molecules might be associated with the development of CHD, independent of other CHD risk factors. PMID- 17135630 TI - Interviews with andrologists (part 2). Interview by Katja Wolski. PMID- 17135629 TI - Relationship of body size and shape to the development of diabetes in the diabetes prevention program. AB - OBJECTIVE: We report the effects of several different measures of body size at baseline on the subsequent development of diabetes. High levels of body fat predict the onset of diabetes, but this association has not been previously reported in a large multiethnic population of overweight or obese people with impaired glucose tolerance. RESEARCH METHODS AND PROCEDURES: Height, weight, waist circumference, hip circumference, and skinfolds were measured at baseline in 3234 participants enrolled in a randomized clinical trial to treat individuals with impaired glucose tolerance with placebo, metformin, or a lifestyle modification program. Cox proportional hazards models were used to assess the effect of baseline body size variables on the development of diabetes. RESULTS: Over an average of 3.2 years in both the placebo and lifestyle groups, baseline waist circumference had the highest or second highest R(2) value for predicting diabetes in both sexes. Cox hazard ratios per 1 standard deviation were 1.43 and 1.49 for men in the placebo or lifestyle groups, respectively, and 1.29 and 1.53 for women in the placebo and lifestyle groups, respectively, adjusted for age and self-reported race/ethnicity. The c-statistic from the receiver operating characteristic curves also favored the waist circumference in men and women in the lifestyle group and men in the placebo group. No components of body size were predictive in the metformin-treated group, and metformin compared with the placebo group was effective in preventing diabetes only in individuals with a BMI > or =35 kg/m(2) or a waist circumference > or =98.0 cm. DISCUSSION: Large waist circumference was a better predictor of risk for developing diabetes than most other measures in the placebo and lifestyle groups. No baseline measure of body size or shape predicted risk of diabetes in the metformin-treated group. PMID- 17135632 TI - Standardization and quality control for determination of fructose in seminal plasma. PMID- 17135633 TI - Relationships between serum hormone levels and semen quality among men from an infertility clinic. AB - Participation rates in epidemiologic studies on semen quality are generally very low, raising concerns as to the potential for selection bias. Since hormones both initiate and maintain spermatogenesis, they may serve as surrogates of semen quality in epidemiologic studies. For this reason, in the present study, we explored the influence and predictive ability of reproductive and thyroid hormones on semen quality among men who were partners in an infertile couple. Between 1999 and 2003, 388 men were recruited from Massachusetts General Hospital Andrology Laboratory for clinical evaluation of fertility status. Fresh semen samples were assessed for quality (concentration, motility and morphology) and the serum levels of hormones, including follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B, sex hormone-binding globulin (SHBG), testosterone, free androgen index, free T4, total T3, and thyroid-stimulating hormone (TSH), were measured. Multiple logistic regression revealed increased odds for below-reference sperm concentration and morphology in men with increased FSH, and decreased odds for below-reference sperm concentration and motility in men with increased inhibin B. When FSH and inhibin B were divided into quintiles, the relationships with sperm concentration showed evidence of a threshold value. However, the ability of specific FSH (10 IU/L) and/or inhibin B (80 pg/mL) cutoff values to predict semen quality was lower than in previous reports. In multiple linear regression analysis, FSH and LH were inversely associated with sperm concentration, motility, and morphology. Inhibin B and free T4 were positively associated with sperm concentration, while there was a suggestive positive association between testosterone and sperm motility. In conclusion, we have found that FSH, LH, inhibin B, testosterone and free T4 levels are associated with human semen parameters. Additional consideration should be given to the utility of serum hormone levels as a surrogate for semen quality in epidemiologic studies in which the collection of semen is difficult due to logistical and/or volunteer rate constraints. PMID- 17135634 TI - Career advice for oncology trainees. PMID- 17135635 TI - Positron emission tomography scans in postchemotherapy seminoma patients with residual masses: a retrospective review from Indiana University Hospital. PMID- 17135636 TI - Dasatinib induces a response in malignant thymoma. PMID- 17135637 TI - Immunohistochemical patterns of reactive microenvironment are associated with clinicobiologic behavior in follicular lymphoma patients. AB - PURPOSE: Recent molecular data have suggested that non-neoplastic cells are powerful modulators that may confer a selective advantage or disadvantage on the outcome of follicular lymphoma (FL) patients. PATIENTS AND METHODS: The prevalence of the principal inflammatory and immune-infiltrated cells was measured immunohistochemically in the tissue of 211 FL patients, and associations were sought with their traditional clinicobiologic characteristics. RESULTS: Our results confirmed the presence of a large number of T lymphocytes (CD4+ and CD8+) and CD57+ cells and, at a moderate level, the presence of TIA-1+ cytotoxic cells, CD68+ macrophages, CD123+ plasmacytoid cells, and FOXP3+ regulatory T cells among the pool of non-neoplastic cells. In addition to the conventional clinical variables, univariate analysis identified a low level of infiltrated CD8+ T lymphocytes as a significantly negative prognostic factor of overall survival. The following significant differences in the abundance of cells of specific and nonspecific immunity were observed in relation to the clinicobiologic features of FL: (1) a reactive microenvironment mainly made up of T lymphocytes and macrophages was significantly associated with a favorable clinical behavior of FL patients; and (2) a reactive microenvironment infiltrated predominantly by CD57+ T cells was associated with a significantly higher frequency of adverse clinicobiologic manifestations such as "B" symptoms and bone marrow involvement. CONCLUSION: Our results demonstrate the existence of two specific patterns in the reactive microenvironment of FL, an immunosurveillance pattern (T lymphocytes and macrophages) and an immune-escape pattern (CD57+ T cells), that were directly associated with the clinicobiologic features of these patients. PMID- 17135638 TI - Quantification of regulatory T cells enables the identification of high-risk breast cancer patients and those at risk of late relapse. AB - PURPOSE: To assess the clinical significance of tumor-infiltrating FOXP3-positive regulatory T cells (TR) in breast cancer patients with long-term follow-up. PATIENTS AND METHODS: FOXP3-positive TR were detected by immunohistochemistry with our new, extensively characterized FOXP3 monoclonal antibody, 236A/E7. Numbers of FOXP3-positive lymphocytes in tissue microarray cores from pure ductal carcinoma in situ (DCIS; n = 62), invasive breast cancer (n = 237) or from comparable areas of normal terminal duct lobular breast tissue (n = 10) were determined. A median cutoff of > or = 15 defined patients with high numbers of TR. RESULTS: TR numbers were significantly higher in in situ and invasive breast carcinomas than in normal breast; invasive tumors have significantly higher numbers than DCIS (P = .001). High numbers of FOXP3-positive TR identified patients with DCIS at increased risk of relapse (P = .04) and patients with invasive tumors with both shorter relapse-free (P = .004) and overall survival (P = .007). High TR numbers were present in high-grade tumors (P < or = .001), in patients with lymph node involvement (P = .01), and in estrogen receptor (ER) negative tumors (P = .001). Importantly, high numbers of TR within ER-positive tumors identified high-risk patients (P = .005). Unlike conventional clinicopathologic factors, high numbers of FOXP3-positive TR can identify patients at risk of relapse after 5 years. CONCLUSION: These findings indicate that quantification of FOXP3-positive TR in breast tumors is valuable for assessing disease prognosis and progression, and that TR are an important therapeutic target for breast cancer. FOXP3-positive TR represent a novel marker for identifying late-relapse patients who may benefit from aromatase therapy after standard tamoxifen treatment. PMID- 17135639 TI - Phase III trial comparing doxorubicin plus cyclophosphamide with docetaxel plus cyclophosphamide as adjuvant therapy for operable breast cancer. AB - PURPOSE: The combination of doxorubicin and cyclophosphamide (AC) is a standard adjuvant chemotherapy regimen. Studies of docetaxel and cyclophosphamide (TC) in metastatic breast cancer (MBC) showed promise in MBC. In 1997, we initiated a randomized adjuvant trial of TC compared with standard-dose AC with a primary end point of disease-free survival (DFS). PATIENTS AND METHODS: Patients were eligible if they had stage I to III operable invasive breast cancer with complete surgical excision of the primary tumor. Between June 1997 and December 1999, 1,016 patients were randomly assigned to four cycles of either standard-dose AC (60 and 600 mg/m2, respectively; n = 510) or TC (75 and 600 mg/m2, respectively; n = 506), administered intravenously every 3 weeks as adjuvant chemotherapy. Radiation therapy (as indicated) and tamoxifen, for patients with hormone receptor-positive disease, were administered after completion of chemotherapy. RESULTS: Both treatment groups (TC and AC) were well balanced with respect to major prognostic factors. Patients were observed through 2005 for a median of 5.5 years. At 5 years, DFS rate was significantly superior for TC compared with AC (86% v 80%, respectively; hazard ratio [HR] = 0.67; 95% CI, 0.50 to 0.94; P = .015). Overall survival rates for TC and AC were 90% and 87%, respectively (HR = 0.76; 95% CI, 0.52 to 1.1; P = .13). More myalgia, arthralgia, edema, and febrile neutropenia occurred on the TC arm; more nausea and vomiting occurred on the AC arm as well as one incident of congestive heart failure. CONCLUSION: At 5 years, TC was associated with a superior DFS and a different toxicity profile compared with AC. PMID- 17135640 TI - Phase II study of cisplatin plus epirubicin salvage chemotherapy in refractory germ cell tumors. AB - PURPOSE: Initial cisplatin (CIS) combination chemotherapy will cure 70% of patients with disseminated testicular cancer. This phase II clinical trial evaluated the combination of CIS plus epirubicin (CIS-EPI) in patients with metastatic germ cell tumors (GCT) not amenable to cure with standard salvage therapy. PATIENTS AND METHODS: Between March 2001 and August 2005, 30 patients with GCT, who had received at least one previous CIS-based regimen, were enrolled. All patients were males, with median age 36 (range, 24 to 45 years). Twenty-one patients (70%) had experienced late relapses (> 2 years). Patients received EPI 90 mg/m2 on day 1 and CIS 20 mg/m2 on days 1 to 5 every 3 weeks for maximum of four cycles. RESULTS: Nineteen (63%) of 30 patients received all four cycles. Toxicity was primarily hematologic: grade 3/4 neutropenia, four patients (one neutropenic fever); two patients had grade 3 thrombocytopenia, and five patients had grade 3/4 anemia. Nonhematologic toxicity was grade 3 acute renal failure in two patients; grade 3 electrolyte wasting in two patients; grade 3 nausea/vomiting in eight patients; grade 3 elevation of aminotransferases in one patient; and grade 3 diarrhea in one patient. There were no occurrences of severe mucositis, cardiotoxicity, or treatment-related deaths. Nine patients achieved a complete remission; seven of these patients remain without evidence of disease at 25+, 27+, 29+, 44+, 45+, 46+, and 48+ months. One patient remains alive with stable pulmonary nodules at 28+ months. CONCLUSION: CIS-EPI is an active regimen in metastatic GCT, with an acceptable toxicity profile. This regimen offers potential for long-term disease-free survival in this population. PMID- 17135641 TI - Docetaxel, estramustine, and 15-month androgen deprivation for men with prostate specific antigen progression after definitive local therapy for prostate cancer. AB - PURPOSE: Androgen-deprivation therapy (ADT) is effective for relapsed prostate cancer, but is rarely curative. The purpose of this trial was to determine the feasibility, toxicity, and prostate-specific antigen (PSA) response of chemotherapy and limited ADT for men with PSA relapse. PATIENTS AND METHODS: Eligible men had an increasing PSA and no metastases after prostatectomy and/or radiation for localized disease. Treatment consisted of four cycles of docetaxel (70 mg/m2) every 21 days and estramustine 280 mg tid on days 1 through 5. After chemotherapy, goserelin acetate and bicalutamide were prescribed for 15 months. RESULTS: Sixty-two patients were enrolled. A complete PSA response (CR) was defined as PSA at or below the assay-specific lower limit. The proportion of patients with CR after chemotherapy, after ADT, and at 1 year after completion of ADT was 53%, 63%, and 36%, respectively. Testosterone was more than 100 ng/dL (median, 250 ng/dL) 1 year after completion of ADT in 97% of patients. Patients with a PSA less than 3.0 ng/mL at enrollment had a significantly longer time to progression (TTP; P = .0004). Of 56 patients who were observed at least 1 year after completion of ADT, 23 (41%) have not experienced progression as of their last follow-up. The median TTP is 34 months from treatment initiation (maximum, 74 months free from progression). CONCLUSION: Chemotherapy plus ADT was feasible for early prostate cancer relapse. Forty-one percent of men who are at least 1 year after completion of ADT with recovered testosterone have not experienced progression. This approach is being tested in a randomized trial with investigation of predictors of response. PMID- 17135642 TI - Plasma Epstein-Barr viral deoxyribonucleic acid quantitation complements tumor node-metastasis staging prognostication in nasopharyngeal carcinoma. AB - PURPOSE: To evaluate the effect of combining circulating Epstein-Barr viral (EBV) DNA load data with TNM staging data in pretherapy prognostication of nasopharyngeal carcinoma (NPC). PATIENTS AND METHODS: Three hundred seventy-six patients with all stages of NPC were studied. Pretreatment plasma/serum EBV DNA concentrations were quantified by a polymerase chain reaction assay. Determinants of overall survival were assessed by multivariate analysis. Survival probabilities of patient groups, segregated by clinical stage (I, II, III, or IV) alone and also according to EBV DNA load (low or high), were compared. RESULTS: Pretherapy circulating EBV DNA load is an independent prognostic factor for overall survival in NPC. Patients with early-stage disease were segregated by EBV DNA levels into a poor-risk subgroup with survival similar to that of stage III disease and a good-risk subgroup with survival similar to stage I disease. CONCLUSION: Pretherapy circulating EBV DNA load is an independent prognostic factor to International Union Against Cancer (UICC) staging in NPC. Combined interpretation of EBV DNA data with UICC staging data leads to alteration of risk definition of patient subsets, with improved risk discrimination in early-stage disease. Validation studies are awaited. PMID- 17135643 TI - Significance of necrosis in grading of oligodendroglial neoplasms: a clinicopathologic and genetic study of newly diagnosed high-grade gliomas. AB - PURPOSE: High-grade gliomas (HGGs; WHO grades 3-4) are highly diverse, with survival times ranging from months to years. WHO 2000 grading criteria for high grade oligodendroglial neoplasms [anaplastic oligoastrocytoma (AOA) and anaplastic oligodendroglioma (AO)] remain subjective, and the existence of grade 4 variants is controversial. PATIENTS AND METHODS: Overall survival (OS) of 1,093 adult patients with a cerebral HGG newly diagnosed between 1990 and 2005 was analyzed by univariate and multivariate models for significance of the following factors: patient age, surgery type, year of diagnosis, endothelial proliferation, necrosis, oligodendroglial histology, treatment center, and chromosome 1p, 19q, 7p (EGFR), and 10q (PTEN) abnormalities by fluorescence in situ hybridization (FISH). RESULTS: Necrosis was a statistically significant predictor of poor OS on univariate and multivariate analyses in AOA but not in AO. Median OS for patients with necrotic AOA (22.8 months) was significantly worse than for patients with non-necrotic AOA (86.9 months; P < .0001) but was better than conventional glioblastomas (9.8 months; P < .0001). In addition to patient age, the following were significant independent prognostic factors (P .001): grade and surgery type for the entire HGG cohort; modified grade for AOA (3 v 4); and modified grade, 1p/19q codeletion status, and oligodendroglial histology for the 586 HGGs analyzed by FISH. CONCLUSION: Stratification of AOA, but not of pure AO, into grades 3 and 4 on the basis of necrosis is prognostically justified and is more powerful than the current approach. Both routine histology and genetic testing provide independent, prognostically useful information. PMID- 17135644 TI - Detrimental effects of tumor progression on cognitive function of patients with high-grade glioma. AB - PURPOSE: There is growing recognition that the primary cause of cognitive deficits in adult patients with primary brain tumors is the tumor itself and more significantly, tumor progression. To assess the cognitive performance of high grade glioma patients, prospectively collected cognitive performance data were analyzed. PATIENTS AND METHODS: We studied 1,244 high-grade brain tumor patients entered onto eight consecutive North Central Cancer Treatment Group treatment trials that used radiation and nitrosourea-based chemotherapy. Imaging studies and Folstein Mini-Mental State Examination (MMSE) scores recorded at baseline, 6, 12, 18, and 24 months were analyzed to assess tumor status and cognitive function over time. RESULTS: The proportion of patients without tumor progression who experienced clinically significant cognitive deterioration compared with baseline was stable at 6, 12, 18, and 24 months (18%, 16%, 14%, and 13%, respectively). In patients without radiographic evidence of progression, clinically significant deterioration in MMSE scores was a strong predictor of a more rapid time to tumor progression and death. At evaluations preceding interval radiographic evidence of progression, there was significant deterioration in MMSE scores for patients who were to experience progression, whereas the scores remained stable for the patients who did not have tumor progression. CONCLUSION: The proportion of high grade glioma patients with cognitive deterioration over time is stable, most consistent with the constant pressure of tumor progression over time. Although other factors may contribute to cognitive decline, the predominant cause of cognitive decline seems to be subclinical tumor progression that precedes radiographic changes. PMID- 17135645 TI - A simple and innovative device to measure arm volume at home for patients with lymphedema after breast cancer. AB - PURPOSE: We designed an arm volumeter specifically for home use based on the water displacement method. The objective of this study was to determine its accuracy and precision, and compare it with a standard volumeter used in lymphedema clinics worldwide. PATIENTS AND METHODS: Using a standard model hospital volumeter and our own device, we took three consecutive measurements of 11 specially cast cylinders, which had known volumes ranging from 10 mL to 4 L, and measurements of both arms of 15 volunteers. RESULTS: Measurements with both volumeters were highly accurate (R2 = 0.9999) when compared with the known volumes of the cast cylinders, and were strongly correlated (R2 = 0.9974) when each arm volume was compared between volumeters. Measurements with our volumeter were more precise both with the cylinders (average standard deviation [SD], 3.2 v 8 mL; P = .0553) and with the arms (average SD, 11.1 v 19 mL; P = .0034). Whereas the standard volumeter is expensive, fragile (acrylic), and prone to leaks, our volumeter is inexpensive, virtually indestructible, leak proof, and suitable for home use. CONCLUSION: Arm volumes can be measured quickly and accurately at home using a simple, inexpensive, and robust device based on water displacement. Readily accessible arm volumetry at home may have widespread influence on the management of lymphedema after breast cancer. PMID- 17135646 TI - Phase III trial comparing supportive care alone with supportive care with oral topotecan in patients with relapsed small-cell lung cancer. AB - PURPOSE: For patients with small-cell lung cancer (SCLC), further chemotherapy is routinely considered at relapse after first-line therapy. However, proof of clinical benefit has not been documented. PATIENTS AND METHODS: This study randomly assigned patients with relapsed SCLC not considered as candidates for standard intravenous therapy to best supportive care (BSC) alone (n = 70) or oral topotecan (2.3 mg/m2/d, days 1 through 5, every 21 days) plus BSC (topotecan; n = 71). RESULTS: In the intent-to-treat population, survival (primary end point) was prolonged in the topotecan group (log-rank P = .0104). Median survival with BSC was 13.9 weeks (95% CI, 11.1 to 18.6) and with topotecan, 25.9 weeks (95% CI, 18.3 to 31.6). Statistical significance for survival was maintained in a subgroup of patients with a short treatment-free interval (< or = 60 days). Response to topotecan was 7% partial and 44% stable disease. Patients on topotecan had slower quality of life deterioration and greater symptom control. Principal toxicities with topotecan were hematological: grade 4 neutropenia, 33%; grade 4 thrombocytopenia, 7%; and grade 3/4 anemia, 25%. Comparing topotecan with BSC, infection grade 2 was 14% versus 12% and sepsis 4% versus 1%; other grade 3/4 events included vomiting 3% versus 0, diarrhea 6% versus 0, dyspnea 3% versus 9%, and pain 3% versus 6%. Toxic deaths occurred in four patients (6%) in the topotecan arm. All cause mortality within 30 days of random assignment was 13% on BSC and 7% on topotecan. CONCLUSION: Chemotherapy with oral topotecan is associated with prolongation of survival and quality of life benefit in patients with relapsed SCLC. PMID- 17135647 TI - Phase II trial of amrubicin for treatment of refractory or relapsed small-cell lung cancer: Thoracic Oncology Research Group Study 0301. AB - PURPOSE: This multicenter, phase II study was conducted to evaluate the activity of amrubicin, a topoisomerase II inhibitor, against refractory or relapsed small cell lung cancer (SCLC). PATIENTS AND METHODS: SCLC patients with measurable disease who had been treated previously with at least one platinum-based chemotherapy regimen and had an Eastern Cooperative Oncology Group performance status of 0 to 2 were eligible. Two groups of patients were selected: patients who experienced first-line treatment failure less than 60 days from treatment discontinuation (refractory group), and patients who responded to first-line treatment and experienced disease progression > or = 60 days after treatment discontinuation (sensitive group). Amrubicin was administered as a 5-minute daily intravenous injection at a dose of 40 mg/m2 for 3 consecutive days, every 3 weeks. RESULTS: Between June 2003 and December 2004, 60 patients (16 refractory and 44 sensitive) were enrolled. The median number of treatment cycles was four (range, one to eight). Grade 3 or 4 hematologic toxicities comprised neutropenia (83%), thrombocytopenia (20%), and anemia (33%). Febrile neutropenia was observed in three patients (5%). Nonhematologic toxicities were mild. No treatment-related death was observed. The overall response rates were 50% (95% CI, 25% to 75%) in the refractory group, and 52% (95% CI, 37% to 68%) in the sensitive group. The progression-free survival, overall survival, and 1-year survival in the refractory group and the sensitive group were 2.6 and 4.2 months, 10.3 and 11.6 months, and 40% and 46%, respectively. CONCLUSION: Amrubicin exhibits significant activity against SCLC, with predictable and manageable toxicities; this agent deserves to be studied more extensively in additional trials. PMID- 17135648 TI - Do parents have the right to refuse standard treatment for their child with favorable-prognosis cancer? Ethical and legal concerns. PMID- 17135649 TI - Efficacy of complementary and alternative medicine therapies in relieving cancer pain: a systematic review. AB - PURPOSE: Despite widespread popular use of complementary and alternative medicine (CAM) therapies, a rigorous evidence base about their efficacy for cancer-related pain is lacking. This is a systematic review of randomized controlled trials (RCTs) evaluating CAM therapies for cancer-related pain. METHODS: RCTs using CAM interventions for cancer-related pain were abstracted using Medline, EMBASE, CINAHL, AMED, and Cochrane database. RESULTS: Eighteen trials were identified (eight poor, three intermediate, and seven high quality based on Jadad score), with a total of 1,499 patients. Median sample size was 53 patients, and median intervention duration was 45 days. All studies were from single institutions, four had sample size justification, and none reported any adverse effects. Seven trials reported significant benefit for the following CAM therapies: acupuncture (n = 1), support groups (n = 2), hypnosis (n = 1), relaxation/imagery (n = 2), and herbal supplement/HESA-A (n = 1, but study was of low quality without control data). Seven studies reported immediate postintervention or short-term benefit of the following CAM interventions: acupuncture (n = 2), music (n = 1), herbal supplement/Ai-Tong-Ping (n = 1), massage (n = 1), and healing touch (n = 2). Four studies reported no benefit of CAM interventions (music, n = 2; massage, n = 2) in reducing cancer pain compared with a control arm. CONCLUSION: There is paucity of multi-institutional RCTs evaluating CAM interventions for cancer pain with adequate power, duration, and sham control. Hypnosis, imagery, support groups, acupuncture, and healing touch seem promising, particularly in the short term, but none can be recommended because of a paucity of rigorous trials. Future research should focus on methodologically strong RCTs to determine potential efficacy of these CAM interventions. PMID- 17135650 TI - Epidermal growth factor receptor gene copy number in esophageal cancer and outcome prediction to gefitinib: does intratumoral heterogeneity matter? PMID- 17135651 TI - Chemotherapy-induced steatohepatitis in colorectal cancer patients. PMID- 17135652 TI - Effects of comorbidity and smoking on the survival of lung cancer patients. PMID- 17135653 TI - Does CTLA4 influence the suppressive effect of CD25+CD4+ regulatory T cells? PMID- 17135654 TI - Problems with up-front randomization in clinical trials. PMID- 17135655 TI - Standard of care for gastric cancer based on meta-analysis? Treading on thin ice or it is very nice! PMID- 17135656 TI - Import and fate of fluorescent analogs of oxidized phospholipids in vascular smooth muscle cells. AB - Lipid oxidation is now thought to be an initiating and sustaining event in atherogenesis. Oxidatively fragmented phospholipids, namely 1-palmitoyl-2 glutaroyl-sn-glycero-3-phosphocholine (PGPC) and 1-palmitoyl-2-(5-oxovaleroyl)-sn glycero-3-phosphocholine (POVPC), present in minimally modified LDL and atherosclerotic lesions, have been reported to elicit a wide range of pathophysiological responses in the cells of the vascular wall. Nevertheless, the question of their potential sites of action and their primary molecular targets remains open. To address this issue, a series of fluorescently labeled analogs, which differ with regard to structure and binding site of the fluorophore, were synthesized and used as tools for studying the uptake, intracellular stability, and distribution of PGPC and POVPC in vascular smooth muscle cells (VSMCs). We demonstrate that in accordance with their lysophospholipid-like structure, these highly similar molecules transferred rapidly either from aqueous phospholipid dispersions or preloaded native LDL into VSMCs, producing disparate fluorescence patterns irrespective of the attached fluorophore. PGPC derivatives were translocated to the lysosomes. In sharp contrast, POVPC analogs were initially captured in the plasma membrane, most likely in consequence of the formation of covalent adducts with free amino and sulfhydryl groups of proteins and phospholipids. LDL internalization is not required for cellular lipid uptake. Collectively, our data provide evidence that oxidized phospholipids, owing to their high exchangeability between lipoproteins and cell membranes, may act within a short time on different cellular sites in VSMCs and affect various lipid and protein components through physical or chemical interactions, which might then serve as starting points for intracellular signaling. PMID- 17135657 TI - No evidence for different dose responses of commercial methionine sources in broilers. PMID- 17135658 TI - Perspectives in chicken genetics and genomics. AB - Poultry science has entered a new era with the completion of a century of investigative studies in chicken genetics, sequencing of the chicken genome, application of genomic tools into systems biology studies, and rapid advances in the development of the statistical theory for application of molecular genetic information in commercial breeding programs. This perspectives paper sets the context for the accompanying series of reviews on chicken genetics and genomics, introduces important issues in the field of poultry molecular genetics, and briefly describes the topics of each of the reviews. PMID- 17135659 TI - Progress from chicken genetics to the chicken genome. AB - The chicken has a proud history, both in genetic research and as a source of food. Here we attempt to provide an overview of past contributions of the chicken in both arenas and to link those contributions to the near future from a genetic perspective. Companion articles will discuss current poultry genetics research in greater detail. The chicken was the first animal species in which Mendelian inheritance was demonstrated. A century later, the chicken was the first among farm animals to have its genome sequenced. Between these firsts, the chicken remained a key organism used in genetic research. Breeding programs, based on sound genetic principles, facilitated the global emergence of the chicken meat and egg industries. Concomitantly, the chicken served as a model whose experimental populations and mutant stocks were used in basic and applied studies with broad application to other species, including humans. In this paper, we review some of these contributions, trace the path from the origin of molecular genetics to the sequence of the chicken genome, and discuss the merits of the chicken as a model organism for furthering our understanding of biology. PMID- 17135660 TI - Strategies to assess structural variation in the chicken genome and its associations with biodiversity and biological performance. AB - A primary goal in the assessment of structural variation in the avian genome is to understand the relationship of this variation with biodiversity and with biological performance. To develop such knowledge, certain essential tools are needed. One set of tools includes the laboratory techniques used to assess molecular genetic variation. The current time is a transitional one for this field, in that the recently sequenced chicken genome will add significantly to the portfolio of existing methods used to identify molecular markers. To most efficiently discover marker-trait associations, the experimental mapping populations must be appropriately designed and the relevant statistical analyses applied. This paper reviews methods for assessment of molecular markers in poultry and their use in the characterization of avian biodiversity and in studies to identify marker associations with biological traits, including important considerations of population structure and statistical analysis. PMID- 17135661 TI - Review of quantitative trait loci identified in the chicken. AB - Methods for mapping QTL are actively used in the chicken to identify chromosomal regions contributing to variation in traits related to growth, disease resistance, egg production, behavior, and metabolic parameters. However, higher resolution mapping and better knowledge of the genetic architecture underlying QTL are needed for successful application of this information into breeding programs. Therefore, this paper summarizes and integrates original, primary QTL studies in the chicken to identify basic information on the genetic architecture of quantitative traits in chickens. The results of this review show several instances of consensus of QTL locations for similar traits from independent studies. Furthermore, the consensus of QTL location for different traits and evidence for QTL with parent-of-origin effect, transgressive alleles, epistatic QTL, and QTL x sex interaction in chicken are presented and discussed. This information can be helpful in identifying genes or mutations underlying the QTL and in the application of genomic information in marker-assisted breeding programs. PMID- 17135662 TI - Effect of arsanilic acid on performance and residual of arsenic in tissue of Japanese laying quail. AB - Three hundred seventy-five 63-d-old laying Japanese quail were randomly distributed into 3 experimental groups (125 birds per group) and fed the following diets for 40 d, with 30 d on the experimental diets, followed by a 10-d withdrawal period: 1) control; 2) 50 mg of 4-arsanilic acid/kg of feed; and 3) 100 mg of 4-arsanilic acid/kg of feed. Each treatment consisted of 5 replicates of 25 birds. During the first 30 d of the experiment, all eggs were recorded, collected, individually weighed daily, and feed consumption was determined every 10 d. Five quail from each replicate in the experiment were euthanized by cervical dislocation at 0, 30, 35, and 40 d. Tissue samples from the liver, kidney, heart, gizzard, and the muscle on the breast and leg were collected for determination of As residue. The feces and eggs at 0, 30, 35, and 40 d of the experiment were selected for determination of As. Results showed that dietary inclusion of 50 and 100 mg/kg of 4-arsanilic acid significantly improved feed utilization and egg production, but the concentration of As in the tissues and feces in groups fed 4-arsanilic acid was higher than in control group. The results of the present study demonstrate that the use of organic As compounds as feed additives in diet is a matter for argument. PMID- 17135663 TI - The effect of an experimental chlorate product on Salmonella recovery of turkeys when administered prior to feed and water withdrawal. AB - Previously, an experimental chlorate product (ECP) has been observed to reduce Escherichia coli and Salmonella infections in swine, cattle, and broilers. The following studies were performed to investigate the effects of different concentrations and durations of administering ECP on crop and ceca Salmonella typhimurium (ST) colonization of turkeys. In 2 separate trials, each conducted with 2 replicates, 15-wk-old turkey toms were challenged with 10(7) to 10(9) cfu of ST. In Experiment 1, toms were administered 0, 0.5, 1.0, 2.0, or 4.0x of ECP (a 1.0x concentration is equivalent to a 15 mM chlorate ion concentration) in the drinking water for 38 h. In Experiment 2, toms were administered a 2x concentration of ECP in the drinking water for 0, 14, 26, or 38 h prior to water withdrawal. All treatments were followed by a 10-h water withdrawal and an 8-h feed withdrawal prior to organ sampling. In Experiment 1, turkeys provided ECP had significantly (P < 0.05) lower populations and incidences of crop (>1.4 log reduction) and ceca (>0.6 log reduction) ST as compared with control birds (2.1 and 0.94 log ST average for all trials, respectively), with little or no additional benefit from administration of higher ECP concentrations. In Experiment 2, toms provided ECP had lower populations of crop (>2.2 log reduction) and ceca (>1.5 log reduction) ST when compared with controls (3.1 and 1.8 log ST, respectively). Again, there appeared to be little benefit in longer administration intervals on quantitative reduction of ST. These experiments suggest that the ECP significantly reduces Salmonella colonization in commercial turkeys when administered prior to feed and water withdrawal. PMID- 17135664 TI - Effects of feed additives and mixed eimeria species infection on intestinal microbial ecology of broilers. AB - Evaluation of digestive microbial ecology is necessary to understand effects of growth-promoting feed. In the current study, the dynamics of intestinal microbial communities (MC) were examined in broilers fed diets supplemented with a combination of antibiotic (bacitracin methylene disalicylate) and ionophore (Coban 60), and diets containing 1 of 2 essential oil (EO) blends, Crina Poultry (CP) and Crina Alternate (CA). Five treatments were analyzed: 1) unmedicated uninfected control; 2) unmedicated infected control; 3) feed additives monensin (bacitracin methylene disalicylate) + monensin (Coban 60; AI); 4) EO blend CP; and 5) EO blend CA. Additives were mixed into a basal feed mixture, and EO were adjusted to 100 ppm. Chicks were infected by oral gavage at 19 d of age with Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Duodenal, ileal, and cecal samples were taken from 12 birds per treatment just before and 7 d after challenge; 2 samples each were pooled to give a final number of 6 samples total; and all pooled samples were frozen until used for DNA extraction. Denaturing gradient gel electrophoresis was used to examine PCR-amplified fragments of the bacterial 16S ribosomal DNA variable region. Results are presented as percentages of similarity coefficients (SC). Dendrograms of PCR amplicon or band patterns indicated MC differences due to intestinal location, feed additives, and cocci challenge. Essential oil blends CP and CA affected MC in all gut sections. Each EO had different effects over MC, and they differed in most instances from the AI group. The cocci challenge caused drastic MC population shifts in duodenal, ileal, and cecal sections (36.7, 55.4, and 36.2% SC, respectively). Diets supplemented with CP supported higher SC between pre- and postchallenge MC (89.9, 83.3, and 76.4%) than AI (81.8., 57.4, and 60.0%). We concluded that mixed coccidia challenge caused drastic shifts in MC. These EO blends modulated MC better than AI, avoiding drastic shifts after a mixed challenge. PMID- 17135665 TI - Influence of a semiochemical analogue on growing performances and meat quality of broilers. AB - Stress in broilers may have severe consequences on the final product quality. A synthetic analogue of uropygial secretion of mother hens was isolated from poultry. This mother hen uropygial secretion analogue (MHUSA) was tested in farm conditions on broilers during 12 wk. The purpose of this trial was to estimate the influence of MHUSA on growing performances, meat characteristics after processing, and stress indicators of broilers. After the 80-d period, birds under treatment were heavier at 3 different weighing ages (P 15I(5) > 6(3) (P < 0.05 and P < 0.01, respectively). These results suggested that genetic selection for disease resistance also directly or indirectly modified the corresponding genetic components that govern the immune and neuroendocrine systems. The genetic lines of chickens may be used as animal models for investigation of the cellular mechanisms of genetic environmental interactions on disease resistance. PMID- 17135670 TI - Role of thyroid hormones, maternal antibodies, and antibody response in the susceptibility to colibacillosis of broiler genotypes. AB - The purpose of this study was to investigate whether differences in susceptibility to colibacillosis are associated with maternal antibodies, antibody response, and alterations in thyroid hormones [triiodothyronine (T3) and thyroxine (T4)] and to investigate the effect of genotype on the changes in T3 and T4 during challenge and antibody response. A challenge experiment was executed in 2 trials. Per trial, 24 chicks per genotype were challenged, and 20 chicks per genotype were controls. At 7 d of age, challenged chicks were intratracheally inoculated with 0.3 mL of Escherichia coli O78K80 and controls with 0.3 mL of PBS. All chicks were euthanized at 14 or 15 d. Thyroid hormone plasma concentrations and E. coli-specific antibody titers (AB) were measured at 7 d (T(3 d7), T(4 d7), and AB(d7)) and 14 or 15 d (change from 7 to 14 or 15 d was analyzed: DeltaT(3), DeltaT(4), and DeltaAB). Susceptibility was defined based on mortality, lesions, growth retardation, and eating behavior. There was a significant effect of challenge on T(3 d7); probably due to eating pattern in association with circadian rhythm. The challenge group was suggested to have functional hypothyroidism relative to the control group, indicating metabolic changes due to the challenge, and it was indicated that an antibody response was elicited. Differences in susceptibility were not significantly related to differences in T(3 d7), T(4 d7), DeltaT(3), or DeltaT(4) or to maternal antibodies (AB(d7)), but the antibody response tended to increase (decreasing DeltaAB) with increasing susceptibility. There were indications of genetic variation in T(4 d7), DeltaT(4), AB(d7), and DeltaAB, but there was no observed effect of genotype on DeltaT(3) and DeltaT(4) during challenge or on the antibody response. Further, there were indications that selection for growth traits has resulted in alterations in DeltaT(4) due to challenge, as indicated by a lower DeltaT(4) in the challenge group relative to the control group for more intensively selected genotypes as opposed to a higher DeltaT(4) for less intensively selected genotypes. PMID- 17135671 TI - Effects of particle size and physical form of diets on mast cell numbers, histamine, and stem cell factor concentration in the small intestine of broiler chickens. AB - The aim of this study was to investigate the hypothesis that particle size and diet form may affect the growth of mast cells and histamine release from the small intestine of broiler chickens. A total of 288, day-old male broiler chicks were randomly allocated to 1 of 4 corn-soy diets in a 2 x 2 factorial design. The factors included particle size (coarse vs. fine) and physical form (mash vs. pellet). The birds were housed in 90 x 60 cm pens containing 12 birds, and each treatment contained 6 replicate pens of birds from d 1 to 22. On d 22, 6 broilers from each treatment were slaughtered. Tissues from the small intestine (duodenum, jejunum, and ileum) were obtained to quantify mast cells using the toluidine blue staining technique. The results showed that mast cells in the jejunum were concentrated in the upper part of the villus in birds fed the coarsely ground mash diet, whereas mast cells were evenly distributed throughout the intestine in birds fed the other 3 diets. The number of mast cells was significantly lower in the duodenum (P = 0.04), jejunum (P < 0.01), and ileum (P = 0.01) of birds fed coarsely ground diets compared with finely ground diets, and there was no difference in mast cell numbers between birds fed mashed or pelleted diets at any site in the intestine. The histamine content (P = 0.02) and stem cell factor concentration (P = 0.03) were markedly lower in the jejunum of birds that were fed coarsely ground diets compared with finely ground diets. The stem cell factor concentration in the duodenum (P < 0.01) and jejunum (P = 0.05) was higher in birds fed pelleted compared with mash diets. The overall results of this experiment suggest that particle size and diet form affect mast cell number and histamine content in the small intestine by regulation of stem cell factor concentration. PMID- 17135672 TI - Age- and breed-dependent adapted immune responsiveness of poultry to intratracheal-administered, pathogen-associated molecular patterns. AB - Immune modulation of poultry by airborne pathogen-associated molecular patterns (PAMP) was studied. White and Brown layer chicks were exposed intratracheally during 5 consecutive days at 7 wk of age with Escherichia coli-derived lipopolysaccharide (LPS), Saccharomyces cerevisiae-derived 1,3 beta-glucan (BGL), a combination of both, or PBS as a control. Six weeks later, birds received similar or crossover PAMP treatments. Body weight (gain), feed conversion, (primary and secondary) specific antibody responses to model antigens, and natural antibody levels were measured. In general, BGL enhanced but LPS exposure decreased primary immune responses at 7 wk of age, whereas both PAMP-enhanced secondary immune responses but decreased primary immune responses at 13 wk of age. Body weight gain and feed conversion at both ages were negatively affected by LPS, especially in White birds, but not by BGL. Pathogen-associated molecular patterns exposure at 7 wk of age also affected Ab responses at 13 wk of age. Birds exposed to a combination of LPS + BGL at 7 wk of age had significantly lower secondary total and IgG Ab responses at 13 wk of age. Birds from both breeds showed enhanced BW gain after exposure to LPS at 13 wk of age, when initially challenged at 7 wk of age with LPS, BGL, or a combined challenge with both. Pathogen-associated molecular patterns exposure at 7 wk of age affected humoral immunity and BW gain at 13 wk of age in a positive (BGL) or negative (LPS) fashion. Repeated exposure to PAMP did not affect Ab responses, but crossover exposure to PAMP in general enhanced Ab responses. Body weight gain was positively affected by repeated exposure but not by crossover exposure, suggesting adaptation of the birds to early PAMP exposure. Our findings suggest that sensitivity of poultry for immune modulation by airborne PAMP differs between ages, is breed-dependent, and is not irreversible of nature. In addition, our data suggest different adaptation to hygienic conditions, both with respect to immune reactivity and BW gain. PMID- 17135673 TI - Chinese herbal ingredients are effective immune stimulators for chickens infected with the Newcastle disease virus. AB - This study was conducted to determine the efficacy of 4 Chinese herbal ingredients (CHI) as immune stimulators for an active vaccine in chickens using both in vitro and in vivo assays. The CHI used were Astragalus polysaccharide (APS), Isatis root polysaccharide (IRPS), Propolis polysaccharide, and Epimedium flavone at various concentrations. Two hundred 14-d-old male White Roman chickens were randomly divided into 10 groups. Chickens in groups 1 to 9 were inoculated with the New-castle disease virus (NDV) strain IV vaccine by intranasal and intraocular administration. Chickens in groups 1 to 8 were also administered subcutaneously on the dorsal region of the neck with 0.5 mL of the corresponding CHI at 2 doses: 29 and 58 mg/kg of BW for APS and IRPS and 7.25 and 14.5 mg/kg of BW for the others, once daily for 3 successive days. In group 9 (CHI-free control) and group 10 (both vaccine- and CHI-free control), chickens were injected with 0.5 mL of physiological saline. New-castle disease virus-specific serum hemagglutination inhibition antibody (Ab) production in immunized chickens was quantified using established methods. The results indicate that a majority of the CHI used at appropriate concentrations were effective in enhancing in vitro proliferation of chick embryo fibroblasts in response to the NDV infection. In vivo administration of CHI to vaccinated chickens (7.25 to 58 mg/kg of BW, depending on type) increased serum anti-NDV hemagglutination inhibition Ab titer concentrations, compared with the administration the NDV alone. For all CHI, a beneficial effect on the Ab production was observed on d 21 after the initiation of the vaccination. On the basis of the in vivo doses used, Propolis polysaccharide and Epimedium flavone were more potent than APS and IRPS in promoting the humoral immune response in the young birds (P < 0.05). Collectively, these findings suggest that appropriate doses of CHI can be used as novel, effective immune stimulators for chickens. PMID- 17135674 TI - Effect of beta-mannanase (Hemicell) on growth performance and immunity of broilers. AB - Two hundred four broilers (1-d-old) were randomly allocated to 4 treatments, each of which had 3 pens of 17 chicks per pen and were used to investigate the effects of beta-mannanase (Hemicell) on growth performance and immunity. The chicks received the same basal diet based on corn-soybean meal and Hemicell was added to the basal diet at 0, 0.025, 0.05, and 0.075%, respectively. Weight of each replicate was determined at wk 0, 3, and 6 of age. There were no significant differences in average feed intake in the 0- to 3-wk and 0- to 6-wk periods, and no differences in serum IgA, or IgG concentrations. However, the addition of Hemicell significantly increased (P < 0.05) weight gain in the 4- to 6-wk and 0- to 6-wk periods. Feed conversion for the 0.025 and 0.05% groups was significantly greater (P < 0.05) than for the control group in the 4- to 6-wk and 0- to 6-wk periods. Hemicell significantly increased (P < 0.05) the serum IgM concentration in 3- and 6-wk-old broilers. Proliferation of T lymphocytes in 6-wk-old broilers for the 0.05% group was also improved (P < 0.05) significantly. The results indicate that Hemicell may improve growth performance and immunity of broilers. PMID- 17135675 TI - The influence of exogenous multienzyme preparation and graded levels of digestible lysine in sunflower meal-based diets on the performance of young broiler chicks two weeks posthatching. AB - The study was conducted to investigate the effect of adding multienzyme preparation (Rovabio Excel AP, Adisseo Asia Pacific Pte Ltd., Singapore) at 2 inclusion levels of sunflower meal (SFM; 20 and 30% of the diets) with 3 levels of digestible Lys (0.8, 0.9, and 1.0%) with and without enzyme in a 2 x 2 x 3 factorial arrangement. Each diet was offered to 4 replicates of 51 one-day-old straight-run Hubbard broiler chicks (n = 2448) in a practical vegetable-based mash diet having 2,750 kcal of ME/kg and 19% CP during 2 wk posthatching (1 to 14 d of age). Feed formulation was based on digestible amino acids, which were calculated from the CP and DM contents of each ingredient using AminoDat 2 (Degussa Corp., Allendale, NJ). The enzyme used in this study was authenticated by the supplier to have minimum level of endo-1,4-beta xylanase (22,000 visco units/g) and endo-1,3(4)-beta glucanase (2,000 AGL units/g) and was added at the rate of 50 mg/kg of finished diet. No significant effect of enzyme or level of SFM was observed on BW gain (BWG), feed intake, or mortality during the experimental period. The BWG and feed:gain for birds fed on 30% SFM with enzyme were comparable to those fed on 20% SFM without enzyme during 1 to 14 d of age. However, enzyme at 20% SFM depressed the BWG (P or = 0.60 (p < 0.05, n = 75). An increase in the proportion of prostate stones (corpora amylacea), stroma, or cancer in relation to healthy glandular tissue increased the measured stiffness. Cancer and stroma had the greatest effect on the measured stiffness. The deeper the sensor was pressed, the greater, i.e., deeper, volume it sensed. Tissue sections deeper in the tissue were assigned a lower mathematical weighting than sections closer to the sensor probe. It is concluded that cancer increases the measured stiffness as compared with healthy glandular tissue, but areas with predominantly stroma or many stones could be more difficult to differ from cancer. PMID- 17135707 TI - Cerebral critical closing pressure estimation from Finapres and arterial blood pressure measurements in the aorta. AB - Estimates of cerebral critical closing pressure (CrCP) and resistance-area product (RAP) are often derived using noninvasive measurements of arterial blood pressure (ABP) in the finger, but the errors introduced by this approach, in relation to intra-vascular measurements of ABP, are not known. Continuous recordings of ABP (Finapres and solid-state catheter-tip transducer in the ascending aorta), cerebral blood flow velocity (CBFV, bilateral Doppler), ECG and transcutaneous CO(2) were performed following coronary catheterization. CrCP and RAP were calculated for each of 12,784 cardiac cycles from 27 subjects using the classical linear regression (LR) of the instantaneous CBFV-ABP relationship and also the first harmonic (H(1)) of the Fourier transform. There was a better agreement between LR and H(1) for the aortic measurements than for the Finapres (p < 0.000,01). For LR there were no significant differences for either CrCP or RAP due to the source of ABP measurement, but for H(1) the differences were highly significant (p < 0.000,03). The coherence functions between either CrCP or RAP values calculated with aortic pressure (input) or the Finapres (output) were significantly higher for H(1) than for LR for most harmonics below 0.2 Hz. When using the Finapres to estimate CrCP and RAP values, the LR method produces similar results to intra-arterial measurements of ABP for time-averaged values, but H(1) should be preferred in applications analysing beat-to-beat changes in these parameters. PMID- 17135708 TI - Vasopressin stimulates Na-dependent phosphate transport and calcification in rat aortic smooth muscle cells. AB - We investigated the effect of arginine vasopressin (AVP) on inorganic phosphate (Pi) transport in A-10 rat aortic vascular smooth muscle cells (VSMCs). AVP time- and dose-dependently stimulated Na-dependent Pi transport in A-10 cells. This stimulatory effect of AVP on Pi transport was markedly suppressed by V1 receptor antagonist. A protein kinase C (PKC) inhibitor calphostin C partially suppressed the stimulatory effect of AVP. The selective inhibitors of c-Jun-NH2-terminal mitogen-activated protein (MAP) kinase (Jun kinase) attenuated AVP-induced Pi transport, but Erk kinase or p38 MAP kinase inhibitors did not. Wortmannin, a phosphatidylinositol (PI) 3-kinase inhibitor, suppressed AVP-induced Pi transport. Rapamycin, a selective inhibitor of S6 kinase, reduced this effect of AVP, while Akt kinase inhibitor did not. The combination of inhibitors for PKC, Jun kinase and PI 3-kinase completely suppressed the AVP-enhanced Pi transport. Furthermore, AVP rescued the VSMC from high phosphate-induced cell death and enhanced mineralization of these cells. In summary, these results suggest that AVP stimulates both Na-dependent Pi transport and mineralization in VSMCs. The mechanism is mediated by the activation of multiple signaling pathways including PKC, PI 3-kinase, S6 kinase and Jun kinase. PMID- 17135709 TI - A case of hepatitis C-associated osteosclerosis in an elderly man. Comment on the article by Tanaka et al. PMID- 17135710 TI - Role of the hyaluronan receptor CD44 during porcine oocyte maturation. AB - Previous our studies have shown that CD44, the principal receptor for hyaluronan, is present on cumulus cells during oocyte maturation. Although hyaluronan-CD44 interaction has been implicated in cumulus expansion and/or oocyte maturation, the full significance of CD44 remains unknown. The objective of the present study was to further investigate the role of CD44 in cumulus expansion and oocyte maturation in pigs. We demonstrate here in that CD44 has a key role in oocyte maturation but not in cumulus expansion. Previous studies have reported the physiological significance of cumulus expansion in oocyte maturation. However, our results suggest that cumulus expansion is a necessary condition for oocyte maturation, but that it is not sufficient on its own. Furthermore, western blot analysis demonstrated that the CD44 of the in vitro-matured cumulus-oocyte complexes (COCs) had a larger molecular weight and more terminal sialic acid, which has been proven to inhibit the hyaluronan-binding ability of the receptor, than the CD44 of the in vivo-matured COCs, indicating that the hyaluronan-CD44 interactions during in vitro maturation might be insufficient compared with those in vivo. The insufficient interactions of hyaluronan-CD44 during in vitro maturation may cause the inferior capacity of fertilization and development of oocytes matured in vitro. PMID- 17135711 TI - Acrosome reaction of mouse epididymal sperm on oocyte zona pellucida. AB - To improve assessment of the acrosome reaction of mouse epididymal sperm, we employed anti-Izumo1 antibody instead of antibodies against acrosomal proteins. The acrosomal states among acrosome-intact, spontaneously acrosome-reacted, truly acrosome-reacted, and probably dead and/or membrane-damaged sperm were clearly distinguished by combined application of anti-Izumo1 antibody, DNA dye Hoechst 33342, and monoclonal antibody MN7 to paraformaldehyde-fixed sperm. When the acrosome reaction of capacitated epididymal sperm on the oocyte zona pellucida was examined using anti-Izumo1 antibody, approximately 20% of sperm bound onto the zona pellucida were acrosome-reacted 30 min after insemination. We also observed the moment of the acrosome reaction of live sperm on the zona pellucida by time-lapse monitoring using fluorescein isothiocyanate-conjugated anti-Izumo1 antibody. PMID- 17135712 TI - Peripartum changes in plasma estrone sulfate and estradiol-17beta profiles associated with and without the retention of fetal membranes in holstein-friesian cattle. AB - The aim of this study was to evaluate the changes in plasma concentrations of estrone sulfate (E(1)S) and estradiol-17beta (E(2)beta) during the peripartum period (from day 10 prepartum to day 1 postpartum) associated with and without retention of fetal membranes (RFM) in Holstein-Friesian cattle (n=42). Plasma samples were analyzed for E(1)S and E(2)beta by ELISA. All parturitions were spontaneous and normal. Of 38 cattle delivering singletons, 29 had no RFM (singleton-normal group) and nine had RFM for more than 12 h (singleton-RFM group). Four cows gave birth to twins, and each twin had its own fetal membrane (FM). Two twinning cows expelled both FMs normally within 12 h (twin-normal group). In the remaining 2 twinning cows (twin-RFM group), the FM was expelled normally for one twin (first), while the FM of the other (second) was retained. There were no significant differences in the E(1)S concentrations or their increments from the concentrations on the preceding day between the normal and RFM groups of singleton cows on any peripartum day. The mean plasma E(2)beta concentrations on each day from day 10 to day 3 prepartum were significantly lower (P<0.05) in the singleton-RFM group compared with the singleton-normal group; however, on days 2 and 1 prepartum, the increments in the E(2)beta concentrations from the concentrations on the preceding days were significantly higher (P<0.05) in the singleton-RFM group than in the singleton-normal group. Thus, the plasma E(1)S concentrations just before parturition may not be associated with RFM. In the cows with RFM, the lower plasma E(2)beta concentrations that were found prior to day 2 prepartum may have been associated with immature placentomes, and the rapid rise in plasma E(2) beta within 1 to 2 days prior to calving may have produced asynchrony of placental and/or fetal maturation in relation to calving, thus resulting in RFM. PMID- 17135713 TI - Conventional freezing of in vitro-produced and biopsied bovine blastocysts in the presence of a low concentration of glycerol and sucrose. AB - The purpose of this study was to develop a practical cryopreservation method for in vitro-produced (IVP) and sex-predetermined bovine blastocysts that will be applicable to direct transfer of the post-thaw embryos. Blastocysts were harvested 7 days after IVF and allocated to either an intact or biopsy group. The cryoprotective solution contained 0.7 M glycerol and 0, 0.05 or 0.1 M sucrose. Slow cooling at a rate of -0.5 C/min was terminated at -25, -30, or -35 C, and rapid cooling in liquid nitrogen was followed. After one-step thawing and dilution, the IVP blastocysts were cultured for 3 days to assess their survival. The post-thaw survival rate of intact blastocysts after termination of slow cooling at -30 C in 0.7 M glycerol plus 0.1 M sucrose (96.2%) was significantly higher than that at -25 C in 0.7 M glycerol alone (44.4%). The post-thaw survival rate of biopsied bovine blastocysts after termination of slow cooling at -25 C in 0.7 M glycerol alone (53.8%) tended to be lower than that at -25 C in 0.7 M glycerol plus 0.05 M sucrose (91.3%) or -30 C in 0.7 M glycerol plus 0.1 M sucrose (92.3%). Thus, addition of a small amount of sucrose to 0.7 M glycerol cryoprotective solution shortened the process of slow cooling for both the intact and biopsied bovine embryos. Judged from the survival levels in vitro after thawing and one-step dilution of embryos (>80%), this is an improved method of cryopreservation for subsequent direct transfer of IVP and biopsied bovine blastocysts. PMID- 17135714 TI - Progesterone plus PMSG Priming in seasonally anovulatory lactating Sarda ewes exposed to the ram effect. AB - The aim of this trial was to evaluate the effectiveness (fertility and lambing) of priming with a single injection of progesterone plus PMSG in anovulatory lactating Sarda ewes subjected to the ram effect (RE) in spring. Thirty ewes (P4 group) were i.m. injected with 30 mg progesterone and 500 IU PMSG 36 h before ram introduction (d 0). This treatment was compared to a 12-day treatment with fluorogestone acetate intravaginal sponges that was followed by injections of 350 IU PMSG upon sponge withdrawal (FGA group, n=30). All ewes responded to RE, showing plasma progestrone concentrations >1 ng/mL between d 6 and 12 (FGA) or 6 and 9 (P4). Eighty-nine percent of the P4 ewes conceived at first ovulation, and 11% conceived following a short estrus cycle. Lambings occurred on d 150.4 +/- 3.9, and the lambing rate was 100%. The fertility of the FGA ewes was 83% for the induced ovulation and was 7% for the second ovulation after a normal cycle. The FGA ewes lambed on d 149.8 +/- 4.4, and the lambing rate was 83%. Two abortions were recorded for the FGA ewes, which had higher prolificacy than the P4 group (2.2 +/- 0.8 vs. 1.8 +/- 0.4, respectively; P<0.05). Both fertility and the lambing rate were high in both groups, with a high degree of estrus synchronization, and there were no significant differences between the groups. We concluded that priming of lactating Sarda ewes in spring with P4+PMSG before RE is an effective and competitive method (cheaper and more practical than FGA+PMSG) of inducing fertile ovulations in these ewes. PMID- 17135715 TI - A novel apoptosis gene identified in the pituitary gland. AB - Although multiple different cancers have been described, it is likely that these tumour types share a small, and common, number of newly acquired functional capabilities. Tumours that arise within the pituitary gland are no exception with respect to these new functional capabilities. Although compelling evidence for self-sufficiency in growth signals is presented, loss of functional tumour suppressor genes by classic mechanisms has not been clearly established. However, and in this context, methylation-mediated or -associated gene silencing, in particular of tumour suppressor genes, has been reported by numerous investigators in this tumour type. More recently, a search for novel genes on the basis of their inappropriate methylation has led to identification of a novel pro apoptotic gene. Its pituitary tumour derivation and role in drug-induced apoptosis resulted in the acronym PTAG (pituitary tumour apoptosis gene) being assigned to this gene. In a model pituitary tumour cell line, AtT20, expression of PTAG per se had no discernible effects on proliferation, cell cycle profile or viability. However, enforced expression was associated with a significantly increased sensitivity to the apoptotic effects induced by bromocriptine challenge. Apoptosis was mediated through caspase activation and associated with DNA fragmentation as determined by TUNEL labelling. Similar findings are also evident in the rodent pituitary cell line, GH3 and our data shows that drugs other than bromocriptine, and that engage characterized receptors, elicit a PTAG augmented apoptotic response. The isolation of novel genes, on the basis of their methylation status, offers a significant advantage with respect to our understanding of tumorigenesis in both the pituitary and other tumour types. The reversal of apparent gene silencing may lead to tumour cell 'sensitisation' to chemo- and radiotherapeutic treatment strategies. PMID- 17135716 TI - Co-expression of tyrosine hydroxylase and GTP cyclohydrolase I in arginine vasopressin-synthesizing neurons of the human supraoptic nucleus demonstrated by laser microdissection and real-time PCR. AB - Tyrosine hydroxylase (TH), the first and limiting enzyme for catecholamine synthesis, has been identified immunohistochemically (IHC) in human neurosecretory neurons where it is found to colocalize with vasopressin (AVP) or oxytocin. TH expression shows striking interindividual variability and appears to depend on neuronal activation. Since GTP cyclohydrolase I (GCHI), the first enzyme for tetrahydrobiopterin synthesis, the essential cofactor of TH, and aromatic L-amino acid decarboxylase (AADC) have so far not been detected in neurosecretory neurons, the functional role of TH in catecholamine synthesis is still questionable. Our purpose was to investigate in postmortem hypothalamus whether GCHI and AADC mRNAs are co-expressed with TH in human AVP-synthesizing neurons. Total RNA was extracted from laser microdissected TH-IHC-identified neurons as well as from dissected parts of the dorsolateral supraoptic nucleus (dl-SON) of 12 control subjects, i.e. without known neurological, psychiatric or endocrinological illness. GCHI, AADC and TH mRNA expression was determined by real-time PCR. Our results showed that GCHI mRNA is co-expressed with TH in almost all cases that had a considerable number of TH-immunoreactive (TH-IR) neurosecretory neurons. A positive correlation was found between TH immunohistochemical intensity and the presence of GCHI mRNA. AADC mRNA expression was detected only in microdissected areas of dl-SON in 2 cases that showed an increased number of TH-IR neurons. The co-expression of GCHI with TH indicates that TH is indeed active in human neurosecretory neurons. The apparent limited expression of AADC indicates that dopamine might be produced in human neurosecretory neurons under activation of the hypothalamoneurohypophyseal system, although the possibility that L-dopa is the final product cannot be excluded. PMID- 17135717 TI - Suppression of adrenocorticotrophic hormone secretion by simultaneous antagonism of vasopressin 1b and CRH-1 receptors on three different stress models. AB - BACKGROUND/AIMS: Corticotrophin-releasing hormone (CRH), adrenocorticotrophic hormone (ACTH) and corticosterone are secreted during stress. These mediators may be involved in anxiety, depression and post-traumatic stress disorder, therefore antagonists have been developed to treat such conditions. METHODS: The non peptide CRH receptor type 1 antagonist CP154,526 and the vasopressin receptor type 1b antagonist SSR149415 were used to suppress the secretion of ACTH induced by ether exposure, forced swimming and restraint in adult male Wistar rats. Doses ranged from 3 to 60 mg/kg s.c. (controls with vehicle) alone or in combination, in varying time schedules to assess the duration and effectiveness of treatments. RESULTS: Stressors increased plasma ACTH by 2.5- to 5-fold in control rats. SSR149415 at doses of 30 mg/kg was more effective at suppressing ACTH secretion after ether exposure and restraint but was ineffective against forced swimming. CP154,526 mildly affected ACTH rise after restraint at doses of 30 mg/kg. The combination of both antagonists at doses of 30 mg/kg effectively blocked the rise in plasma ACTH in all three stresses. The drug effects lasted less than 6 h. CONCLUSION: We demonstrated for the first time that simultaneous blockade of both vasopressin 1b and CRH-R1 receptors effectively abolish the ACTH response to physical and psychological stress modalities. PMID- 17135718 TI - A woman with facial papules and pulmonary nodules. PMID- 17135719 TI - A patient with progressive multiple myeloma treated successfully with arsenic trioxide after allogeneic bone marrow transplantation. AB - Multiple myeloma (MM) is an incurable progressive disease. Many therapeutic options are available to delay progression, including autologous and allogeneic bone marrow transplantation. At advanced stages, MM is often refractory to treatment. We report a heavily pretreated patient with graft-versus-host disease after bone marrow transplantations, treated at a terminal stage with a modified protocol for arsenic trioxide (ATO). This patient with poor clinical status tolerated the treatment very well. He had a remarkable clinical response and achieved complete remission. The mechanisms of ATO are presented and the potential role of ATO for MM is discussed. PMID- 17135720 TI - Tyramide signal amplification: an enhanced method for immunohistochemistry on methyl-methacrylate-embedded bone marrow trephine sections. AB - The detection of cellular antigens in bone marrow sections depends on the method of embedding, the nature of antigen and antibody, antigen retrieval techniques and the sensitivity of the immunohistochemical method. This study evaluated a fluorescyl-tyramide-enhanced immunostaining method on methyl-methacrylate embedded bone marrow trephine biopsies. Compared with the standard immunostaining technique, fluorescyl-tyramide-enhanced immunostaining showed a superior signal intensity on methyl- methacrylate-embedded bone marrow trephines. Thus, the fluorescyl-tyramide-based immunostaining method, while retaining specificity, is highly sensitive and provides optimal staining results for a broad panel of antibodies commonly used for the routine diagnosis and classification of hematological disorders. PMID- 17135721 TI - Serum levels of vitamin D and haemostatic factors in healthy subjects: the Tromso study. AB - BACKGROUND: Receptors for vitamin D have been found in various tissues, including the vascular endothelium. The role of vitamin D in the haemostatic process is uncertain, but in vitro studies may indicate a pro-fibrinolytic effect. METHODS: Two hundred and six subjects (105 males) were included in the study. The relations between indices of calcium metabolism and haemostatic factors [tissue plasminogen activator antigen (tPA Ag), plasminogen activator inhibitor 1 (PAI 1), prothrombin fragment 1+2, activated factor VII and total factor VII coagulant activity] and high-sensitivity C-reactive protein (HS-CRP) were examined. RESULTS: There were significant and negative correlations between serum 25(OH) vitamin D and PAI-1 and tPA Ag, and between serum 1,25(OH)2 vitamin D and tPA Ag and HS-CRP. In a multiple linear regression model with age, gender, body mass index and smoking status as covariables, only the relation between 25(OH) vitamin D and tPA Ag was significant. There were no significant relations between any of the haemostatic factors tested and serum parathyroid hormone. CONCLUSION: It appears that the serum level of vitamin D is related to fibrinolytic activity and to the integrity of the vascular endothelium, but the clinical importance of this observation remains to be determined. PMID- 17135722 TI - Plasma homocysteine and hematological factors in patients with venous thromboembolic diseases in Kuwait. AB - BACKGROUND: Venous thromboembolic disease (VTE) is a common cause of morbidity in Kuwait, but the risk factors have not been studied. Hyperhomocysteinemia has been suggested as one of the risk factors. We postulate that hyperhomocysteinemia acts synergistically with hematological variables to increase VTE risk. This study evaluates the roles of hyperhomocysteinemia and hematological variables in patients with VTE. METHODS: We measured fasting plasma total homocysteine (tHcy), activated protein C resistance, protein C (PC), protein S (PS) and antithrombin (AT) in 201 patients with VTE and 166 healthy controls. We also measured factor VIII, factor II, lupus anticoagulant, anticardiolipin, serum vitamin B12, folate, creatinine, lipid profile, glucose, full blood count and red cell folate. The glomerular filtration rate (GFR) was calculated from creatinine. RESULTS: When patients on warfarin were excluded, 13.1% of patients (18 out of 137) had a deficiency in PC, 16.8% (23 out of 137) had a deficiency in PS, and when patients on heparin were excluded, 8.3% of patients (14 out of 168) had low AT. Spearman's rank correlation analysis showed that tHcy had significant correlations with age, creatinine and PS, and significant inverse correlations with GFR, high-density lipoprotein cholesterol and serum folate. Partial correlation analysis after correcting for age and sex showed that tHcy retained a significant correlation with creatinine, GFR and serum folate. Binary logistic regression analyses of the determinants of hyperhomocysteinemia included age, creatinine, GFR and serum folate. Multivariate logistic regression analysis showed significant association of tHcy with VTE (OR = 5.6; p < 0.0001) in the presence of known risk factors for VTE. CONCLUSION: We conclude that elevated tHcy is a significant risk factor for the development of VTE, and therefore, it should be included in the workup for patients at risk of VTE, but the determinants of tHcy should be kept in mind. PMID- 17135723 TI - Late relapses in acute promyelocytic leukaemia. AB - From January 1988 to December 1997, among 53 acute promyelocytic leukaemia patients in 1st complete remission (CR) after 5 years from diagnosis, we observed 5 late relapses (9.4%) after 60, 61, 71, 101 and 155 months from diagnosis; 3 of those late relapses (7.7%) occurred among 39 patients previously treated with all trans-retinoic acid. An involvement of the mastoid occurred in 3/5 patients (60%), compared with 2/32 patients (6.3%) at an early relapse (p < 0.02). As to the treatment of the late relapse, 1 patient received all-trans-retinoic acid alone followed by allogeneic transplantation and 4 patients were treated according to the GIMEMA 0191 protocol. All patients achieved a 2nd CR and are still alive: 4 in the 2nd molecular CR after 6, 33, 34 and 115 months; 1 relapsed after 15 months and is now in the 3rd CR. In conclusion, a late relapse occurred in a sizeable fraction of acute promyelocytic leukaemia patients: the high rate of ear involvement might be explained considering the ear as a 'disease sanctuary'. PMID- 17135724 TI - Pilot remission induction therapy with idarubicin, plus an intensified dose of ara-C and priming with granulocyte colony-stimulating factor for acute myeloid leukemia. AB - BACKGROUND: The sensitization of leukemic cells with hematopoietic growth factors can enhance the cytotoxicity of chemotherapy in acute myeloid leukemia (AML). Intensified remission induction (RI) therapy can also improve the treatment results for AML. Therefore, the current trial attempted to evaluate the efficacy and toxicity of granulocyte colony-stimulating factor (G-CSF) priming and a dose intensification of Ara-C in RI chemotherapy for AML. METHODS: A total of 29 patients with newly diagnosed AML received G-CSF-priming RI chemotherapy consisting of idarubicin (12 mg/m2, days 1-3), G-CSF (150 microg/m2, days 3-8) and Ara-C (500 mg/m2, b.i.d., days 4-8), and the outcomes were compared with those of a historical group treated with a standard regimen consisting of idarubicin (12 mg/m2, days 1-3) and Ara-C (100 mg/m2, days 1-7). RESULTS: There was no difference in sex, age, subtype and cytogenetic risk between the two groups. The complete remission rate and treatment-related mortality were 72 and 17% for the G-CSF-primed group (p = 0.89) and 71 and 10% for the historical group (p = 0.32), respectively. The time to neutrophil recovery (25 vs. 24 days, p = 0.17) and platelet recovery (24 vs. 23 days, p = 0.23) did not differ significantly between the two groups. Similarly, the duration of fever was not significantly different (5 vs. 7 days, p = 0.58). Thirteen patients (45%) experienced fever and 5 patients (17%) manifested skin rashes during the G-CSF priming. After a median follow-up of 336 days, the 1-year overall survival, disease-free survival and event-free survival rates were 72 vs. 63% (p = 0.83), 74 vs. 56% (p = 0.059) and 53 vs. 38% (p = 0.32), respectively. CONCLUSION: The sensitization of leukemic cells with growth factors and dose intensification seem to be clinically applicable means to enhance the efficacy of RI chemotherapy only in selected patients with AML, thereby warranting further studies focusing on specific subgroups of AML patients. PMID- 17135725 TI - HHV-8-associated lymphoma: state-of-the-art review. AB - During the first decade after the discovery of primary effusion lymphoma (PEL), sporadic and serial reports suggested that Kaposi-sarcoma-associated herpesvirus/human-herpesvirus-8 (KSHV/HHV-8)-associated lymphomas in their liquid and solid presentation are clinically distinct, representing part of the spectrum of PEL. In HIV-seropositive patients with serous effusions, these solid lymphomas were reported before the development of an effusion lymphoma and following resolution of PEL. More recently, solid lymphomas not associated with lymphomatous effusion have been found in HIV-seropositive and HIV-seronegative patients. Despite the diversification in the clinical presentation of KSHV/HHV-8 associated lymphomas, most cases demonstrated a similar morphology, immunophenotype and KSHV/HHV-8 viral status. PMID- 17135726 TI - Comparison of human herpes virus 8 related primary effusion lymphoma with human herpes virus 8 unrelated primary effusion lymphoma-like lymphoma on the basis of HIV: report of 2 cases and review of 212 cases in the literature. AB - BACKGROUND: Primary lymphomatous effusion is a rare lymphoma that arises in the body cavity and has a peculiar proliferative form, lacking a tumor. This primary lymphomatous effusion includes human herpes virus 8 (HHV8)-related primary effusion lymphoma (PEL) and HHV8-unrelated PEL-like lymphoma. We attempted to clarify the nature of the primary lymphomatous effusion. METHODS: Using 'PEL' and 'body cavity-based lymphoma' (BCBL) as key words, reports written in English were collected from PubMed. Primary lymphomatous effusion was defined as BCBL with primary effusion and without tumor at onset. Adding our 2 PEL-like lymphoma cases, each case was studied as to the patients' and lymphomas' characteristics, therapy and survival time. Moreover, each item was compared among four groups according to the presence of HHV8 and HIV. RESULTS: In 214 cases investigated, there was no difference in proliferation, but an apparent difference in age, gender, phenotype, effectiveness and prognosis among the four groups. CONCLUSIONS: Both PEL and PEL-like lymphoma are thought to be characterized by a peculiar proliferation, regardless of the presence of HHV8. Dividing PEL or PEL like lymphoma into two subgroups on the basis of HIV presentation might also be appropriate. PMID- 17135727 TI - Reliability study on the Japanese version of the Clinician's Interview-Based Impression of Change. Analysis of subscale items and 'clinician's impression'. AB - BACKGROUND/AIMS: The Japanese version of the Clinician's Interview-Based Impression of Change plus Caregiver Input (CIBIC-plus J) consists of 3 subscales: Disability Assessment of Dementia Scale (DAD), Behavioral Pathology in Alzheimer's Disease Rating Scale (Behave-AD), and Mental Function Impairment Scale (MENFIS), as well as the Clinician's Global Impression of Change (CGIC). While the interrater reliability of CGIC has already been reported, that of the 3 subscales has not. The aim of the present report was to examine the reliabilities of the subscale items and investigate their relationships with CGIC. METHODS: Eleven raters who were clinical physicians watched videotapes of 20 patients with Alzheimer's disease, completed the CIBIC-plus J assessment form, and assigned a CGIC score to the patients. Reliability was assessed using the kappa coefficient. RESULTS: The kappa coefficient of the subscale items was in most instances higher than that of CGIC (0.453) and substantial reliability was observed. The Spearman rank correlation that was calculated between CGIC and the total score change of items was very high for MENFIS (0.990) and DAD (0.910), and moderate for Behave AD items (0.576). The incidence of comments by the raters was highest for MENFIS (89%), followed by DAD (70%). The incidence was low for Behave-AD items (48%). CONCLUSION: Based on the results, it is concluded that DAD, Behave-AD, and MENFIS are necessary constituents of CIBIC-plus J, and indispensable for the reliability of CGIC. PMID- 17135728 TI - Use of sedation for routine diagnostic upper gastrointestinal endoscopy: a European Society of Gastrointestinal Endoscopy Survey of National Endoscopy Society Members. AB - BACKGROUND/AIMS: Sedation rates may vary among countries, depending on patients' and endoscopists' preferences. The aim of this survey was to investigate the rate of using premedication for routine diagnostic upper gastrointestinal (UGI) endoscopy in endoscopy societies, members of the European Society of Gastrointestinal Endoscopy (ESGE). METHODS: We evaluated a multiple-choice questionnaire which was e-mailed to representatives of national endoscopy societies, which are members of the ESGE. The questionnaire had 14 items referring to endoscopy practices in each country and the representatives' endoscopy units. RESULTS: The response rate was 76% (34/45). In 47% of the countries, less than 25% of patients undergo routine diagnostic UGI endoscopy with conscious sedation. In 62% of the responders' endoscopy units, patients are not asked their preference for sedation and do not sign a consent form (59%). Common sedatives in use are midazolam (82%), diazepam (38%) or propofol (47%). Monitoring equipment is not available 'in most of the endoscopy units' in 46% (13/28) of the countries. Though they were available in 91% of the national representatives' endoscopy units, they are rarely (21%) used to monitor unsedated routine diagnostic UGI endoscopy. CONCLUSIONS: In about 50% of ESGE-related countries, less than 25% of patients are sedated for routine diagnostic UGI endoscopy. Major issues to improve include availability of monitoring equipment and the use of a consent form. PMID- 17135729 TI - Effects of a preload on reduction of food intake by GLP-1 in healthy subjects. AB - BACKGROUND/AIMS: Glucagon-like peptide-1 (GLP-1) inhibits food intake in animals and humans. Whether GLP-1 interacts with other satiety signals to modulate food intake is unknown. We investigated therefore in healthy volunteers the potential interactions of GLP-1 with signals from the stomach in regulating food intake. METHODS: Three sequential, double-blind, crossover studies were performed in male subjects: (1) 12 subjects underwent four experiments (preloads) 20 min before meal intake; (2) 12 volunteers received intravenous (i.v.) GLP-1 (0.9 pmol/kg/min) or saline; (3) subjects received i.v. GLP-1 or saline (control) together with a preload of either 400 ml water or 400 ml protein shake. The effect of these treatments on food intake and feelings of hunger was quantified. Subjects were free to eat and drink as much as they wished. RESULTS: GLP-1 induced a reduction in food and calorie intake (p < 0.005) compared to controls. If combined with a protein preload, the inhibitory effect of GLP-1 on food intake was markedly increased (p < 0.001). Furthermore, a decrease in hunger feelings and an increase in satiety feelings was documented. CONCLUSION: GLP-1 interacts with signals from the stomach to modulate energy intake in humans. The signal is only initiated by nutrient-based distension, but not with gastric distension of the fundus alone. PMID- 17135730 TI - Herpesviruses in patients with drug hypersensitivity syndrome: culprits, cofactors or innocent bystanders? PMID- 17135731 TI - Acne and 'mild' adrenal hyperplasia. A short critical review. PMID- 17135732 TI - Need and greed in dermatology: disease and outcome measures in dermatological healthcare planning. AB - Assessing dermatological needs in the population and our ability to meet them is a challenge. The choice of outcome measures is central in dermatological healthcare planning. The concept of ill health and its relationship to dermatological outcome measures is described. Outcome measures used should provide information on these issues in order to gain insight into genuine needs and illness behavior in the population and to ensure quality of healthcare. Nevertheless, the dilemma of assigning priorities in healthcare planning remains; outcome measures in dermatology should be perceived as appropriate by patients as well as by caregivers and policy makers. It is important to stimulate research to develop and refine person-focused outcome measures. Their use is essential to assess dermatological health needs and to understand illness behavior. The information they can provide can enable provision and evaluation of healthcare quality and contribute to the challenge of making appropriate priority decisions in healthcare. PMID- 17135733 TI - Expression pattern of chemokine receptors and chemokine release in inflammatory erythroderma and Sezary syndrome. AB - BACKGROUND: Erythroderma can be caused by inflammatory dermatoses or cutaneous T cell lymphoma. Even if chemokines and their receptors are involved in the skin selective lymphocyte recruitment, their role in inflammatory erythroderma is yet unclear. OBJECTIVE: To evaluate the chemokine release (TARC, MDC, IP-10) and to define the expression pattern of Th1- (CCR5, CXCR3) and Th2-related (CCR4) chemokine receptors in inflammatory erythroderma and Sezary syndrome (SS). MATERIALS AND METHODS: Flow cytometry has been carried out on both circulating and skin-infiltrating T lymphocytes; serum chemokine levels have been evaluated using ELISA techniques. RESULTS: CCR4, CCR5 and CXCR3 were expressed on about 40% of peripheral blood lymphocytes and on the majority of skin-infiltrating lymphocytes in the inflammatory erythroderma patients, whereas the leukemic CD4+CD26- subpopulation in SS was characterized by a high CCR4 expression without a concurrent increase in CCR5 or CXCR3. TARC, MDC and IP-10 serum levels were significantly increased in both erythrodermic and SS patients. CONCLUSIONS: Our results confirm that SS is a Th2 disorder with a selective expression of CCR4, whereas inflammatory erythroderma shares an overexpression of both Th1- and Th2 related chemokine receptors, suggesting an activation of different pathways driving reactive lymphocytes to the skin. PMID- 17135734 TI - Incontinence induces stratum corneum vulnerability and impairs the skin barrier function in the perianal region. AB - BACKGROUND: Incontinence leads to a reduced skin barrier function, while also increasing the risk of skin breakdown. AIM: To investigate the perianal skin barrier function of elderly patients with or without incontinence. METHODS: We compared the skin barrier function by investigating the skin hydration, the skin pH, the amount of ceramide and by making a dermatological skin inspection of the perianal region in patients with and without incontinence. RESULTS: Twenty-one incontinent patients (70.0%) showed a glossy skin, which indicates impairment of the skin barrier while none of the patients in the continence group did so. In the incontinence group, those who had a glossy skin showed a significantly greater skin pH and total amount of ceramide than those with a normal skin (p = 0.0099, p = 0.0179, respectively). CONCLUSIONS: These results demonstrate that a glossy skin may be a useful indicator of a reduced perianal skin barrier function. PMID- 17135735 TI - Surgery for foreign body reactions due to injectable fillers. AB - BACKGROUND: An increasing number of soft tissue fillers have been introduced to the beauty market and these filler substances are widely used as non-toxic, non immunogenic and relatively harmless injectable alternatives to surgical rejuvenation. Generally, facial fillers are injectable - or surgically insertable - products that are used to fill up the volume loss in the aging face. Depending on bioavailability, chemical composition and degradation, fillers can be classified as temporary or permanent, organic or inorganic and autologous or heterologous. OBJECTIVE: A plethora of new products has swamped the beauty market since face rejuvenation has become socially acceptable as well as affordable to a wider population, but adverse reactions cannot be excluded. We present 4 patients with complications after injection of facial fillers [including Artecoll(polymethylmethacrylate microspheres), Restylane (hyaluronic acid), DermaLive (hyaluronic acid plus acrylic hydrogel particles) and Newfill (polylactic acid)] and surgical correction. RESULTS: Surgical intervention led to good aesthetic and functional results after multiple unsuccessful conservative therapies. CONCLUSION: We recommend that only physicians familiar with the injection techniques and the biological and chemical characteristics of the various injectable products should perform such interventions. Especially permanent fillers should be used with utmost reticence in cosmetic surgery and we would recommend their application only in reconstructive procedures. Additionally, documentation and reporting of all adverse effects must be mandatory. PMID- 17135736 TI - Bacteraemia in patients with hidradenitis suppurativa undergoing carbon dioxide laser surgery: detection and quantification of bacteria by lysis-filtration. AB - BACKGROUND: Hidradenitis suppurativa (HS) is a cicatrising and persistent disease of apocrine gland-bearing areas in adults. The severity of this condition varies from a few suppurating lesions to widespread, disabling disease. The aetiology is obscure, but suggested contributory factors include a genetic predisposition, comedones occluding the pilosebaceous apparatus, bacterial infections, and hormonal factors. Treatment consists mainly of surgery, while medical therapies serve principally as adjunct therapy. OBJECTIVES: The aim of the study was to determine the number and type of bacteria circulating in the bloodstream in patients with HS undergoing surgical treatment with a carbon dioxide laser stripping-secondary intention technique. METHODS: Twenty-one patients (20 females and 1 male, mean age 36, range 20-55 years) were included in the study. One blood sample (8.3 ml) was taken before surgery, one during the operation and the last one 10 min after surgery. Five healthy persons (all females, mean age 36, range 23-48 years) not undergoing any operation were used as the controls. The blood was cultured by a lysis-filtration technique which had been shown to be very sensitive. Since the filter catches the microorganisms and colonies are formed during culturing, the number of bacteria in the samples is easily determined. RESULTS: In 6 patients, all samples were negative, which indicates that the method of surgery itself caused no spread of bacteria from the lesions. Bacterial growth in the first blood sample was found in 9 patients, from the second sample in 10 and from the third one in 6. In 1 patient, bacteria were detected in three samples. At least 12 bacterial species were identified. The dominating bacteria were coagulase-negative staphylococci of which most were subtyped as Staphylococcus warneri. Among the anaerobic microorganisms, Propionibacterium acnes and P.granulosum were the most frequently isolated bacteria. The bacterial findings in the blood samples accord well with the results from a previous study in which cultures were taken from the deep parts of the HS lesions. In the 5 controls, no microbial growth was detected. CONCLUSION: The carbon dioxide laser stripping technique caused no additional spread of bacteria into the bloodstream. The evaluation of cultures containing microorganisms from normal skin flora is controversial. Since the bacteria encountered in this study are in close agreement with the findings in cultures from the deeper parts of HS lesions they seem to be relevant. The growth of bacteria in the first blood sample taken before surgery may indicate that some of these patients have bacteria continuously circulating in their blood. PMID- 17135737 TI - Benefits from the use of a pimecrolimus-based treatment in the management of atopic dermatitis in clinical practice. Analysis of a Swiss cohort. AB - BACKGROUND: Controlled studies established the efficacy and good tolerability of pimecrolimus cream 1% for the treatment of atopic dermatitis but they may not reflect real-life use. OBJECTIVE: To evaluate the efficacy, tolerability and cosmetic acceptance of a pimecrolimus-based regimen in daily practice in Switzerland. METHODS: This was a 6-month, open-label, multicentre study in 109 patients (55% > or = 18 years) with atopic dermatitis. Pimecrolimus cream 1% was incorporated into patients' standard treatment protocols. RESULTS: The pimecrolimus-based treatment was well tolerated and produced disease improvement in 65.7% of patients. It was particularly effective on the face (improvement rate: 75.0%). Mean pimecrolimus consumption decreased from 6.4 g/day (months 1-3) to 4.0 g/day (months 3-6) as disease improved. Most patients (74.1%) rated their disease control as 'complete' or 'good' and 90% were highly satisfied with the cream formulation. CONCLUSION: The use of a pimecrolimus-based regimen in everyday practice was effective, well tolerated and well accepted by patients. PMID- 17135738 TI - Efficacy results of a 52-week, randomised, double-blind, safety study of a calcipotriol/betamethasone dipropionate two-compound product (Daivobet/Dovobet/Taclonex) in the treatment of psoriasis vulgaris. AB - BACKGROUND: The calcipotriol/betamethasone dipropionate two-compound product is safe and effective in the short-term treatment of psoriasis. OBJECTIVE: The primary objective was to investigate the safety of two treatment regimens involving use of the two-compound product over 52 weeks. The efficacy results are presented here. METHODS: Six hundred and thirty-four patients were randomised double-blind to treatment (once daily, when required) with either: 52 weeks of two-compound product (two-compound group), 52 weeks of alternating 4-week periods of two-compound product and calcipotriol (alternating group), or 4 weeks of two compound product followed by 48 weeks of calcipotriol (calcipotriol group). RESULTS: There was a trend towards a difference between treatments from the overall treatment effect for the percentage of satisfactory responses for each patient during the study (p = 0.071). This appeared to be due to the comparison of the two-compound and calcipotriol groups (p = 0.025). CONCLUSION: There was a trend towards the efficacy of the two-compound product used for up to 52 weeks being better than that of 4 weeks of the two-compound product followed by 48 weeks of calcipotriol. PMID- 17135739 TI - Psoriasis therapy in real life: the need for registries. AB - The introduction of new therapeutic options for the management of psoriasis is a challenge for the dermatology community, and new tools are needed to face this challenge. This article argues for the establishment of profession-based registries to collect solid, long-term data on the safety and effectiveness of different psoriasis treatment regimens. Managed by health care professionals, registries will be most successful if they enroll patients based on indications for treatment rather than on drugs given. This protects the evaluation process from commercial influences and allows a fair comparison of old- versus new generation psoriasis treatments. In contrast to the patients in a registry who receive care in the natural clinical setting, subjects in randomized clinical trials (RCTs) are selected according to study criteria and may therefore not reflect the experience of patients in clinical practice. It is possible that particular risks and opportunities in the real patient population may therefore go undetected in RCTs. PMID- 17135740 TI - Prurigo pigmentosa successfully treated with low-dose isotretinoin. AB - BACKGROUND/AIMS: Prurigo pigmentosa (PP) is an uncommon inflammatory disease with pruritic reddish papules, papulovesicules or vesicules that are symmetrically localized on the trunk and nape, accompanied by reticular hyperpigmentation. Currently available therapeutic options seem somewhat unsatisfying. Herein, we report an 18-year-old female with PP successfully treated with low-dose isotretinoin. METHODS: The patient presented with a symmetrical pruritic eruption on the lateral sides of her trunk with erythematous papules on a hyperpigmented background. Based on the typical clinical and associated histological findings, the patient was diagnosed as PP and a treatment with low-dose isotretinoin 0.3 mg/kg/day (20 mg/day) was started. RESULTS: After a total of 3 months of 20 mg/day isotretinoin therapy, all erythematous macules and papules resolved and the residual pigmentation had almost disappeared. CONCLUSION: Low-dose isotretinoin is not only adequate for the improvement of erythematous lesions, it also helps resolve the reticular hyperpigmentation of PP. Further studies are needed to observe the efficacy of isotretinoin in the treatment of PP. PMID- 17135741 TI - Echinococcosis: a possible etiology in psoriatic disease. AB - We aimed to investigate seroprevalence of Echinococcus granulosus in patients with psoriasis to determine a possible etiologic role, since both echinococcosis and psoriasis are defined as T cell-mediated diseases. Forty psoriatic patients and 50 age- and sex-matched control subjects were included in the study. IgG specific ELISA was used to determine seropositivity. E. granulosus-specific IgG antibodies were found to be positive in 17/40 (42.5%) of the patients with psoriasis and in 11/50 (22%) of the control subjects (p = 0.008). Our results suggest that echinococcosis might be one of the causative pathogens in the etiopathogenesis of psoriasis in highly endemic regions. PMID- 17135742 TI - 1st congress of the International Dermoscopy Society. PMID- 17135743 TI - Drug-induced hypersensitivity syndrome due to mexiletine hydrochloride associated with reactivation of human herpesvirus 7. AB - It has been suggested that reactivation of human herpesvirus 6 (HHV-6) infection may be involved in the pathogenesis of drug-induced hypersensitivity syndrome. We report a 45-year-old Japanese man who developed a generalized papuloerythematous rash, fever, hepatitis, lymphadenopathy and lymphocytosis with an increased number of atypical lymphocytes. He was diagnosed with DIHS due to mexiletine hydrochloride based on laboratory data, results of a patch test and the clinical course of his complaint, and was treated with systemic steroids. In order to determine whether HHV-6 or -7 was associated with the patient's disease, serological assays and PCR were carried out. Significant increases in antibody titers against HHV-6 and -7 were observed from day 12 to 24. From PCR analysis, none of the peripheral blood mononuclear cells or skin tissue samples contained HHV-6 DNA. All samples, however, were found to contain HHV-7 DNA. Reactivation of HHV-7 could be responsible for drug-induced hypersensitivity syndrome. PMID- 17135744 TI - A case of primary cutaneous nasal type NK/T-cell lymphoma and review of the literature. AB - INTRODUCTION: Cutaneous T-cell lymphoma subtypes are now better identified thanks to progress in immunohistochemistry. This article describes a new case of primary cutaneous natural killer/T-cell lymphoma of nasal type (NKTL-NT) and reviews 18 other cases of this rare neoplasm. CASE REPORT: A 79-year-old man presented with a 3-cm nodular tumor of the left leg occurring on a primary chronic lymphedema of the legs. The lesion was CD56+, CD3 intracytoplasmic+, CD45+ and Epstein-Barr virus+. A comprehensive workup including CT scan and bone marrow biopsy was negative and a diagnosis of NKTL-NT with a primary cutaneous involvement was made. The patient was free of disease under multi-agent chemotherapy after 24 months of follow-up. DISCUSSION: After reviewing 18 other cases of primary cutaneous NKTL-NT, we conclude that the prognosis of these lymphomas is usually poor. However, limited cutaneous forms have a longer median survival than extracutaneous variants. PMID- 17135745 TI - Anti-TNF-alpha therapy in childhood pustular psoriasis. PMID- 17135746 TI - Clinicopathologic features of systemic contact dermatitis from ethylenediamine in cetirizine and levocetirizine. PMID- 17135747 TI - How to save botulinum toxin A in the treatment of focal axillary hyperhidrosis. PMID- 17135748 TI - Cholinergic urticaria and hypohidrosis: a clinical reappraisal. PMID- 17135749 TI - Voluminous fetal chylous ascites: a case of complete spontaneous prenatal regression. AB - We report a case of isolated voluminous fetal ascites discovered during a routine fetal ultrasound at 22 weeks' gestation. Analysis of the fluid showed it to be chyliform and ruled out other causes of fetal ascites. Regular bimonthly ultrasound scans monitored its progressive diminution and then its disappearance. Examination showed the child to be normal at birth. Though the literature indicates that prognosis is usually favorable for infants with isolated fetal chylous ascites, spontaneous regression during pregnancy is an uncommon finding. PMID- 17135750 TI - Antibodies to trophoblast antigens HLA-G, placenta growth factor, and neuroD2 do not improve detection of circulating trophoblast cells in maternal blood. AB - OBJECTIVES: Non-invasive prenatal diagnosis using circulating fetal trophoblast cells has been challenging due to lack of a reproducible trophoblast-specific antibody. We investigated the use of three trophoblast cell-specific antibodies, HLA-G, placenta growth factor, and neuroD2, for the isolation of trophoblast cells from the maternal circulation. METHODS: Trophoblast cells were isolated by density centrifugation from maternal blood samples (gestational age 10-20 weeks, n = 9). All women were carrying a male fetus. Following immunocytochemical staining with the trophoblast-specific antibodies, fluorescent in situ hybridization was performed, to verify whether any stained cells were indeed fetal. RESULTS: The HLA-G antibody had a ubiquitous staining pattern, which was not specific for trophoblast cells. Neither the placenta growth factor nor the neuroD2 antibodies were able to identify any trophoblast cells. Following fluorescent in situ hybridization, no male cells were detected on any of the slides. CONCLUSION: The antibodies used in this study were unable to improve detection of trophoblast cells in the maternal circulation. PMID- 17135751 TI - Antibiotic treatment of intra-amniotic infection with Ureaplasma urealyticum. A case report and literature review. AB - BACKGROUND: Ureaplasma urealyticum is the most common pathogen cultured from the amniotic cavity of women with preterm labor and intact membranes. However, experience with antibiotic eradication of the pathogen in this setting is limited. CASE: We report a case of U. urealyticum isolation from the amniotic cavity of a woman with preterm labor at 27 weeks gestation. The patient was treated with erythromycin base for 1 week, followed by fluoroquinolones and clindamycin for 10 days. A healthy neonate was delivered after spontaneous labor began at 33 weeks. The cultures obtained from the placenta and membranes were sterile, but the histopathology of the placenta revealed acute chorioamnionitis with acute funisitis. CONCLUSION: We suggest that intra-amniotic colonization with Mycoplasma spp. remote from term may be managed expectantly, and that therapy with fluoroquinolones and/or clindamycin may be an appropriate choice. PMID- 17135752 TI - Hypothetical role of prostaglandins in the onset of preterm labor after fetal surgery. AB - Preterm labor is one of the most important factors limiting the advancement of fetal surgery programs. While prostaglandins (PGs) have long been indicated as the key factor in the initiation of labor in humans, there is significant evidence showing that the chorionic membrane acts as a powerful barrier between the decidua/myometrium and amniotic PGs during normal pregnancy. After either open or endoscopic fetal surgery the imperfect, non-hermetical closure of the chorion permits leakage of PGs from the amnionic sac, allowing them to reach the decidua and myometrium. The surgical wound in the chorionic barrier could be the major factor involved in preterm labor and delivery after human fetal surgery. PMID- 17135753 TI - Pharmacokinetics of oral cefatrizine in pregnant and non-pregnant women with reference to fetal distribution. AB - OBJECTIVE: To investigate the effect of gestation on the pharmacokinetics of orally administered beta-lactams, choosing cefatrizine as the model antibiotic. SETTING: A tertiary teaching hospital. DESIGN: Prospective study. METHODS: In 20 women with affected fetuses, 17 by beta-thalassemia major and 3 with congenital malformations, termination of gestation between 19 and 24 weeks was induced by intra-amniotic administration of prostaglandin F(2)(alpha). Pharmacokinetics of cefatrizine in maternal and fetal blood were studied after the administration of three 1 g doses of oral cefatrizine, every 12 h. Twenty female non-pregnant volunteers consisted the control group. RESULTS: Gestation was found to decrease substantially both cefatrizine oral bioavailability and maximum serum plasma concentration (42.8 and 44.5%, respectively) but increased elimination half-life. This effect can be attributed to a substantial increase of the apparent volume of distribution of cefatrizine in relation to a moderate increase of clearance that occurs during pregnancy. Fetal serum cefatrizine levels were lower for the first few hours after administration and then exceeded the corresponding maternal ones. CONCLUSIONS: Our results indicate that gestation decreases the oral bioavailability of cefatrizine. A delay in the maternal drug elimination compared to non-pregnant controls was more pronounced in the fetus. PMID- 17135754 TI - Evolving indications for the EXIT procedure: the usefulness of combining ultrasound and fetal MRI. AB - The EXIT procedure (EX utero Intrapartum Treatment) encompasses a multidisciplinary approach to situations in which airway obstruction is anticipated. Uteroplacental circulation is maintained to avoid neonatal hypoxemia while intubation is attempted. Not only is it useful in congenital diaphragmatic hernia with intrauterine tracheal occlusion, but new indications have been proposed. We present two cases in which EXIT procedure was adopted (huge cervical mass with tracheal compression and a highly vascularized cephalocervical mass) for the same purpose on different grounds. Our two cases stress once more the importance of combining fetal ultrasound and magnetic resonance imaging in the characterization of cervical masses and its usefulness in programming the procedure with a multidisciplinary team. PMID- 17135755 TI - Prenatal diagnosis of fetal exencephaly associated with amniotic band sequence at 17 weeks of gestation by fetal magnetic resonance imaging. AB - We report a fetus with exencephaly diagnosed by fetal magnetic resonance imaging (MRI) at 17 weeks of gestation. Fetal ultrasound performed at 13 and 17 weeks of gestation suggested occipital encephalocele. However, the fetal MRI done at 17 weeks of gestation showed exencephaly and suggested amniotic bands as the cause. By providing early and precise information regarding the abnormality and the possible etiology, the fetal MRI enabled us to provide the couple and their families with accurate information regarding the low recurrence risk of this condition. PMID- 17135756 TI - Alpha-1-antitrypsin and IgA in serial meconium and faeces of healthy breast-fed newborns. AB - BACKGROUND: Meconium is a series of layers formed in the foetal intestine from the 12th week of gestation. High content of meconial alpha-1-antitrypsin (AAT), decreasing within the first several days of extrauterine life appears to reflect the meconium clearance of the gut. At birth, IgA is not present in the meconium and breast-fed infants receive this antibody postnatally with human milk. The aim of the study was to determine changes in AAT concentrations, functional activity of that inhibitor expressed as trypsin inhibitory capacity (TIC) and IgA concentration in serial meconium and faeces, as endogenous biochemical markers discriminating between faeces portions formed in intrauterine and extrauterine life periods of healthy breast-fed newborns. METHODS: A group of 24 healthy breast-fed newborns delivered by spontaneous labour were studied prospectively during the first 4 days of postnatal life. AAT and IgA concentrations in the newborn's meconial and faecal samples and IgA concentration in mother's milk samples taken on the third day after delivery, were determined by radial immunodiffusion. TIC was assessed using BAPNA (N-benzoyl-DL-arginine-p nitroanilide). RESULTS: The medians (range) of AAT concentrations in milligrams per gram of dry meconium or faeces were: 68.8(29.2-138.4) (day 1), 56.9 (30.8 112.8) (day 2), 26.2 (6.8-80.7) (day 3), and 6.6 (1.4-27.1) (day 4). The medians (range) of TIC in milligrams of trypsin/g dry mass of meconium or faeces were: 0.76 (0.33-1.79) (day 1), 0.44 (0.17-1.08) (day 2), 0.16 (0.03-0.56) (day 3), and 0.03 (0-0.11) (day 4). The median (range) of IgA concentration in mothers' milk was 715 mg/dl (420-890). IgA was absent in meconium portions from the first day of life while on the successive days the medians (range) of IgA concentration in mg/g dry mass of meconium and faeces were as follows: 0 (0-2.90) (day 2), 2.50 (1.10-9.60) (day 3), 7.05 (4.10-30.60) (day 4). On day 4 of extrauterine life a negative correlation was found between AAT and IgA concentrations in faeces of the newborns (r = -0.46) and a positive correlation was seen between IgA concentrations in faeces and milk (r = 0.93). CONCLUSIONS: Analyses of the systematic decrease in AAT and increase of IgA concentration in serial portions of meconium and faeces over the first days of extrauterine life of breast-fed newborns can date newborn's faeces portions formed during intrauterine and extrauterine maturation. AAT deposited in foetal intestine is an active antiprotease. PMID- 17135757 TI - Are placental chorionic capillary nucleated red blood cell counts useful compared to umbilical cord blood tests? AB - OBJECTIVE: To compare measurement of fetal nucleated red blood cell (NRBC) counts in paired histologic samples of the placenta and umbilical cord bloods. METHOD: Forty-five randomly selected pregnancies had two determinations of the NRBC count. A sample of umbilical venous blood had a NRBC count measured and sections of the placenta were examined for their villous capillary NRBC counts. RESULTS: Umbilical venous blood had NRBC/100 white blood cell counts ranging from 0 to 67. Paired evaluation of placental tissue had NRBC counts of 0-5 with 60% being zero compared to 8% zero counts in cord blood. There was no correlation between the paired counts (R(2) = 0.04). CONCLUSION: Umbilical cord blood provides different information on fetal NRBC count than does histologic study of the placenta. PMID- 17135758 TI - Is TW3 height prediction more accurate than TW2? Preliminary data. AB - BACKGROUND/AIMS: Skeletal maturation is considered a reliable variable in evaluating the 'tempo' of growth. It is important in the diagnosis of endocrinological diseases, in chronic diseases, in hormonal therapy follow-up and in computing height prediction for prognostic and therapeutic purposes. It is also used when chronological age is not available for minors without known birth dates. There are different methods to evaluate skeletal maturation and height prediction. The Tanner-Whitehouse (TW) method 2 (TW2) has been considered to be the most useful method so far, and has recently been updated with modified height prediction equations (TW2-Mark II). TW3 is the newest method. The aim of this study is to evaluate whether TW3 is more accurate in the assessment of height prediction than TW2-Mark II in a sample of healthy north Italian subjects. METHODS: Anthropometrical data were collected as part of a survey in 1977-1978 in Turin. The sample involved 1,384 healthy children. The children, now adults, have been traced and recalled to measure their final height in order to test height prediction reliability. At present, we have collected 118 adult heights. RESULTS: According to the TW2 method 40% of the males had a height prediction error larger than +/- residual SD (4.1 cm), and with TW3 this was 32.9%. The female height prediction error with TW2 was larger than +/- residual SD (3.6 cm) in 29.2% of girls, and the same value was found with TW3. CONCLUSION: According to our preliminary data, TW3 does not represent any real progress. PMID- 17135759 TI - Final height in patients with idiopathic short stature and high growth hormone responses to stimulation tests. AB - Children with idiopathic short stature (ISS) may have normal or increased growth hormone (GH) responses to provocation tests and achieve a final height (FH) below -2.0 standard deviation score (SDS) if untreated. FH of subjects with high stimulated GH levels has not been studied in detail. AIM: It was the aim of this study to analyse FH in ISS patients with high GH peak responses to the provocation test. PATIENTS AND METHODS: We studied 16 patients (9 pre-pubertal) with ISS and a GH peak >or=40 mU/l to insulin-induced hypoglycaemia. The patients were recalled at age 19.7 +/- 2.5 years for measurement of FH when blood samples were obtained for serum insulin-like growth factor (IGF)-I, IGF binding protein 3, acid-labile subunit and GH binding protein measurements. GH bioactivity was determined using the Nb2 bioassay. RESULTS: FH was -3.1 +/- 1.0 SDS, being significantly lower than target height (TH). At FH, IGF-I levels were within -1.5 and +1.5 SDS for age and sex in 10 patients and higher than +1.5 SDS in 6 patients. IGF binding protein 3, acid-labile subunit, GH binding protein levels and GH bioactivity values were normal. SUMMARY: These data suggest that patients with ISS and high GH levels during a GH stimulation test may have a more compromised FH. The association of severe ISS with a peak GH >40 mU/l might suggest a degree of insensitivity for the GH-IGF-I axis. PMID- 17135760 TI - Characterization of dog allergens Can f 1 and Can f 2. 1. Preparation of their recombinant proteins and antibodies. AB - BACKGROUND: Recombinant dog allergens, rCan f 1 and rCan f 2, and their antibodies are good tools for the characterization of dog allergens in order to develop modern therapeutic and preventive methods for dog allergy. METHODS: In this study, cDNA was synthesized from the mRNA of dog salivary glands and cloned into the pGEX4T vector. rCan f 1 and rCan f 2 containing glutathione S transferase were prepared by an Escherichia coli expression system. The antibodies against the recombinant allergens were prepared in rabbit. The serum of patients with dog allergy was evaluated by ELISA and immunoblot, using the recombinant allergens, goat anti-human immunoglobulin (Ig) E (epsilon) labeled with biotin, and enzyme-labeled streptavidin. The binding of IgE in the serum of patients with dog allergy to dog saliva as a natural antigen was determined in the presence or absence of dog saliva, rCan f 1 and rCan f 2 as competitors. The anaphylactic potential of rCan f 1 and rCan f 2 was evaluated. The body temperature of the mice sensitized with rCan f 1 and rCan f 2 was monitored after intravenous injection of the allergens. The passive cutaneous anaphylaxis reaction was examined for rCan f 1 and rCan f 2. Dog salivary glands, dog saliva and dog hair/dander extracts were analyzed with antibodies by means of an immunoblot assay. The expression of the mRNA of Can f 1 and Can f 2 was verified in various dog tissues by reverse transcription polymerase chain reaction. RESULTS: The E. coli expression system revealed the yield of rCan f 1 and rCan f 2 in 36 and 30 mg/l of culture. The molecular weights of rCan f 1 and rCan f 2 were 18 and 20 kDa in SDS-PAGE, respectively. rCan f 1 and rCan f 2 were found to bind to specific IgE in the serum of dog allergy patients. The binding of IgE in the patient serum for dog saliva was partially inhibited in the presence of rCan f 1 and rCan f 2. These recombinant allergens showed positive signals in passive cutaneous anaphylaxis reaction and induced anaphylactic shock in the mouse model, resulting in a decrease in body temperature. The polyclonal rabbit antibody for rCan f 1 bound to a protein of 20 kDa in the salivary gland, saliva and hair/dander extracts of dogs. The rabbit antibody for rCan f 2 bound to proteins in the saliva and the hair/dander extracts. The proteins possessed a molecular weight of 22/ 23 kDa. Reverse transcription polymerase chain reaction showed the presence of mRNA expression of Can f 1 and Can f 2 not only in the salivary gland but also in dog skin. A clear expression of Can f 2 mRNA was observed in dog skin. CONCLUSIONS: The recombinant allergens and antibodies for Can f 1 and Can f 2 are available for immunological and biochemical characterization of dog allergens. The molecular weight of the natural Can f 1 and Can f 2 in dog saliva and hair/dander extracts showed a higher molecular weight than that of rCan f 1 and rCan f 2. The significance of dog skin as the tissue producing dog allergens, especially Can f 2, should be considered in further studies. PMID- 17135761 TI - Characterization of dog allergens Can f 1 and Can f 2. 2. A comparison of Can f 1 with Can f 2 regarding their biochemical and immunological properties. AB - BACKGROUND: The major dog allergens, Can f 1 and Can f 2, are members of the lipocalin protein family. The characterization of both dog allergens is still not complete. Their deduced amino acid sequences indicate the presence of three cysteine residues, probably connected with a disulfide bridge. We compared the biochemical and immunological properties of Can f 1 with those of Can f 2 using gel filtration, electrophoresis, and immunological assays. METHODS: The rCan f 1, rCan f 2 and dog salivary proteins containing natural Can f 1 and Can f 2 were analyzed by HPLC gel filtration. The recombinant Can f 1 (rCan f 1) and rCan f 2 were analyzed by native and sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) with or without reduction. The binding ability of rabbit IgG purified by protein G affinity chromatography from the antiserum against rCan f 1 and rCan f 2 was examined after a reduction in the recombinant allergens. The immunological cross-reaction between rCan f 1 and rCan f 2 was examined by an enzyme-linked immunosorbent assay (ELISA) using the rabbit IgG against rCan f 1 and rCan f 2. The cross-reaction of human IgE in the serum of a patient with dog allergy between rCan f 1 and rCan f 2 was also analyzed by competitive ELISA. RESULTS: The molecular weights of rCan f 1 and of rCan f 2 were 18 and 21 kDa, respectively, using SDS-PAGE under reducing conditions, but the natural Can f 1 and Can f 2 were separated by HPLC gel filtration into fractions containing proteins of 31 and 34 kDa, respectively. rCan f 1 and rCan f 2 migrated as multiple bands (30-100 kDa) in native PAGE in the presence or absence of a reductant. The molecular weights of natural Can f 1 and of Can f 2 were 20 and 23 kDa, respectively, in SDS-PAGE under reducing conditions. The ability of rabbit IgG to bind to rCan f 1 and rCan f 2 increased after the reduction of the recombinant allergens. The rabbit IgG against rCan f 1 bound to rCan f 2. Cross reaction of human IgE was observed between rCan f 1 and rCan f 2. CONCLUSIONS: In the native and recombinant forms, Can f 1 and Can f 2 possessed a dimer structure under natural (non-reduced) condition. The dimers of Can f 1 and of Can f 2 were not built with a disulfide bridge but by non-covalent association. Cleavage of a disulfide bond of rCan f 1 and rCan f 2 increased the ability of binding of rabbit IgG to the allergens. The cross-reactivity of rabbit IgG and human IgE between rCan f 1 and rCan f 2 indicates that the same epitope(s) was present in Can f 1 and Can f 2. PMID- 17135762 TI - Human eosinophils have an intact Smad signaling pathway leading to a major transforming growth factor-beta target gene expression. AB - BACKGROUND: There is a paradoxical finding that eosinophils are frequently accumulated at the sites of allergic inflammation where transforming growth factor (TGF)-beta, a negative regulator of eosinophil survival, is upregulated; however, eosinophil accumulation is persistent. We thus hypothesized that eosinophils might have aberrant TGF-beta signaling and be unresponsive to TGF beta. To test the hypothesis, we examined the expression and function of Smad proteins, which are central mediators for TGF-beta signaling, in human eosinophils. METHODS: Eosinophils were isolated from the peripheral blood of normal donors, and the expression and activation of endogenous Smad proteins were examined by reverse transcription polymerase chain reaction and Western blotting. The Smad function in the transcription of the major TGF-beta target gene Smad7 was investigated using a dominant negative form of Smad3. The effect of TGF-beta on eosinophil survival was then evaluated by a cell viability assay using normal and asthmatic eosinophils. RESULTS: Human eosinophils expressed mRNAs and proteins of TGF-beta typeI and type II receptors, Smad2, Smad3 and Smad4. TGF beta induced the phosphorylation of Smad2 in eosinophils, which was blocked by SB431542, an inhibitor of TGF-beta type I receptor kinase. A dominant negative Smad3 protein suppressed TGF-beta-induced Smad7 mRNA expression in eosinophils. Finally, TGF-beta prevented granulocyte macrophage colony-stimulating factor- or interferon-gamma-mediated survival of eosinophils obtained from asthmatic patients as well as normal subjects. CONCLUSION: Human eosinophils have an intact Smad signaling pathway leading to a major TGF-beta target gene expression. Thus, eosinophils might become resistant to TGF-beta only in in vivo circumstances. PMID- 17135763 TI - Treatment and outcomes in patients with asthma and allergic rhinitis in the United kingdom. AB - BACKGROUND: Since allergic rhinitis in asthma patients is associated with worse asthma control, the treatment of the comorbid condition may improve outcomes. METHODS: A 1-year retrospective study using the UK Mediplus database (2001-2004) included asthmatic patients aged 15-55 with allergic rhinitis. Patients starting therapy based on the Global Initiative for Asthma guidelines, defined as an increase in inhaled corticosteroids (high-dose inhaled corticosteroids, hdICS), or the addition of montelukast (ICS+MON) or long-acting beta-agonists (ICS+LABA) to ICS, were studied. Univariable and multiple logistic regressions evaluated asthma-related outcomes. RESULTS: Among 2,596 asthma and allergic rhinitis patients, 83.2% initiated ICS+LABA, 12.1% hdICS and 4.7% ICS+MON. The mean age was 34 years and 60% were female. ICS+MON patients had more moderate-severe asthma (p = 0.04). Approximately 84% of the ICS+LABA patients experienced an asthma control failure compared to 50% in the other groups (p < 0.0001). The proportions of patients requiring treatment change were 73.8, 22 and 27.3% in the ICS+LABA, hdICS and ICS+MON groups, respectively (p = 0.001). Asthma-related resource use was similar among all groups. The ICS+MON group received fewer mean prescriptions for oral corticosteroids (p = 0.024) than the other groups (p = 0.026). CONCLUSIONS: In asthma and allergic rhinitis, treatment with ICS+MON or hdICS was associated with lower rates of asthma control failure and fewer treatment changes than the ICS+LABA group. MON users also required fewer oral corticosteroids. PMID- 17135764 TI - Genotypes and haplotypes of CCR2 and CCR3 genes in Japanese cedar pollinosis. AB - Whole genome scan analyses have revealed that the chromosomal region 3p21.3, which contains a gene cluster of the CC chemokine receptor, is possibly critical for the pathogenesis of allergic inflammation. Japanese cedar pollinosis is mediated by a type I allergy and induces seasonal rhinitis and conjunctivitis in humans as the most common form of hay fever in spring in Japan, although the candidate genes for cedar pollinosis remain to be elucidated. We sequenced CCR1, CCR2, CCR3, CCR5, and CCXCR1 using the PCR restriction fragment length polymorphism method in subjects with cedar pollinosis and controls. We found 8 polymorphisms of A111G, Arg127Cys and Arg252Gln in CCXCR1, T885C in CCR1, Val64Ile and T780C in CCR2, T51C in CCR3 and Arg223Gln in CCR5. The transmission disequilibrium test using 60 children with pollinosis and their parents and an association study using unrelated adult subjects (151 patients and 157 controls) showed a significant association of 64Ile in CCR2 and 51C in CCR3 with cedar pollinosis. The frequency of haplotype 64Ile/780C/51C in pollinosis was significantly higher than in controls. Our results suggest that CCR2 and CCR3 genes are candidate genes for Japanese cedar pollinosis. PMID- 17135765 TI - Epigallocatechin-3-gallate inhibits secretion of TNF-alpha, IL-6 and IL-8 through the attenuation of ERK and NF-kappaB in HMC-1 cells. AB - BACKGROUND: Epigallocatechin-3-gallate (EGCG) is a major form of tea catechin and has a variety of biological activities. In the present study, we investigated the effect of EGCG on the secretion of TNF-alpha, IL-6 and IL-8, as well as its possible mechanism of action by using the human mast cell line (HMC-1). METHODS: EGCG was treated before the activation of HMC-1 cells with phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore (A23187). To investigate the effect of EGCG on PMA+A23187-stimulated HMC-1 cells, ELISA, Western blot analysis, electrophorectic mobility shift assay and luciferase assay were used in this study. RESULTS: EGCG (100 microM) inhibited PMA+A23187-induced TNF-alpha, IL-6 and IL-8 expression and production. EGCG inhibited the intracellular Ca(2+) level. EGCG attenuated PMA+A23187-induced NF-kappaB and extracellular signal regulated kinase (ERK1/2) activation, but not that of c-Jun N-terminal kinase or p38 mitogen-activated protein kinase. CONCLUSION: EGCG inhibited the production of TNF-alpha, IL-6 and IL-8 through the inhibition of the intracellular Ca(2+) level, and of ERK1/2 and NF-kappaB activation. These results indicate that EGCG may be helpful in regulating mast-cell-mediated allergic inflammatory response. PMID- 17135766 TI - Response to comparison of solutions for sublingual immunotherapy by Claus Bachert. PMID- 17135767 TI - A new equation for estimating renal function using age, body weight and serum creatinine. AB - BACKGROUND: Many formulas have been developed to estimate glomerular filtration rate (GFR). The aim of our study was to propose a new, more reliable equation. METHODS: The study considered 530 subjects (training sample) with M/F 280/250, age 57.1 +/- 17.4, creatinine clearance (CrCl) 55.2 +/- 38.2 (range 2.1-144.0) for the development the new equation. A linear model was used to describe Cr production using serum Cr (sCr), age, and body weight (BW) as variables: (CrCl + b(4)) . sCr = b(1) - (b(2) . age) + (b(3) . BW) subsequently estimating parameter values by linear least squares, with CrCl as the dependent variable, and 1/sCr, age/sCr, BW/sCr as independent variables. CrCl = {[69.4 - (0.59 . age) + (0.79 . BW)]/sCr} - 3.0 (males) and {[57.3 - (0.37 . age) + (0.51 . BW)]/sCr} - 2.9 (females). A 229-patient renal failure validation sample with M/F 166/63, age 53.0 +/- 14.8, GFR 32.0 +/- 14.3 (range 4.3-69.8), assessed using iohexol Cl, was considered to compare the Cockcroft-Gault (C-G) and MDRD formulas with the new equation for estimating GFR. RESULTS: The mean % error in GFR estimated by the new equation (+2.3 +/- 28.3%) was better than with the C-G and MDRD formulas (+5.2 +/- 30.1% and -11.4 +/- 25.9%, respectively, p < 0.0005 and p < 0.0001), and so was the mean absolute % error, bordering on statistical significance (19.8 +/- 20.3 vs. 21.1 +/- 22.0 and 22.4 +/- 17.3, p = 0.08 and p < 0.005). The precision was also better (RMSE = 7.89 vs. 8.02 and 9.13). The Bland-Altman test showed no GFR over or underestimation trend (measured +/- predicted GFR/2 vs. % error, R2 = 0.001). CONCLUSIONS: The new equation appears to be at least as accurate as the C-G and MDRD formulas for estimating GFR. PMID- 17135768 TI - Glucocorticoid-induced diabetes mellitus: prevalence and risk factors in primary renal diseases. AB - BACKGROUND/AIMS: In patients with primary renal diseases the current knowledge of hyperglycemia associated with corticosteroid therapy is limited. We therefore examined the prevalence and risk factors of glucocorticoid-induced diabetes mellitus (DM) in primary renal diseases. METHODS: Patients were recruited with primary renal diseases who were started on corticosteroids between April 2002 and June 2005. In patients with DM, an impaired fasting glucose level and/or positive urinary glucose analyses before corticosteroids therapy were excluded. RESULTS: During corticosteroid therapy (initial dose: prednisolone 0.75 +/- 0.10 mg/kg/day), DM was newly diagnosed in 17 (40.5%) of 42 patients. All of the 17 patients were diagnosed as having DM by postprandial hyperglycemia at 2 h after lunch, although they had normal fasting blood glucose levels. Age (OR 1.40, 95% CI 1.06-1.84) and body mass index (OR 1.87, 95% CI 1.03-3.38) were determined as independent risk factors for glucocorticoid-induced DM. CONCLUSION: Over 40% of patients with primary renal disease developed DM during treatment with corticosteroids. A high age and high body mass index are the independent risk factors for glucocorticoid-induced DM. 24-hour urinary glucose analyses and postprandial plasma glucose are useful for detecting glucocorticoid-induced DM. PMID- 17135769 TI - Renal cell carcinoma in children: experience of a single center. AB - OBJECTIVE: To evaluate the clinical features and outcome of children with renal cell carcinoma (RCC). PATIENTS AND METHODS: Eleven patients with RCC who were diagnosed between 1972 and 2004 were retrospectively analyzed. Clinical features, histopathology, treatment regimens and outcomes of the patients were evaluated. RESULTS: The male/female ratio was 3:8, with a median age of 10 years. The stage distribution was as follows: 3 patients in stage I, 1 patient in stage II, 3 patients in stage IIIb, and 4 patients in stage IV. Five of 7 patients with stage II-IV received an actinomycin D-based regimen, one received a cisplatin-based regimen, and the other was given 5-fluorouracil (5-FU). In the last patient, interferon-alpha was given in combination with 5-FU; 1 of the patients on the actinomycin D regimen received interferon-alpha as well. All of the stage I patients are alive without disease. Three patients with stage IIIb, stage IV and stage II disease are alive without disease 8, 14 and 26 years after their diagnosis, respectively. The other stage IV and stage IIIb patients died of the disease. CONCLUSION: Nephroureterectomy is the main treatment modality, and it is sufficient for stage I patients. For patients with stage II-IV RCC, interferon alpha and/or actinomycin D-based chemotherapy is the treatment of choice. PMID- 17135770 TI - Prevalence and severity of anaemia in patients with type 2 diabetic nephropathy and different degrees of chronic renal insufficiency. AB - BACKGROUND/AIM: Type 2 diabetes mellitus is the single most common cause of chronic kidney disease (CKD); however its real impact on renal anaemia has not been established. The aim of this study was to evaluate whether onset, severity, and prevalence of anaemia during the course of CKD is different between type 2 diabetic and non-diabetic patients. METHODS: We enrolled 281 patients with: (1) type 2 diabetes and no CKD (n = 75); (2) type 2 diabetes plus CKD (n = 106), and (3) CKD without type 2 diabetes (n = 100). According to K/DOQI guidelines, the patients with renal insufficiency (i.e., those with a glomerular filtration rate <60 ml/min) were subgrouped into three tertiles of CKD: (1) stage 3 (creatinine clearance 60-30 ml/min); (2) stage 4 (creatinine clearance 29-15 ml/min), and (3) stage 5 (creatinine clearance <15 ml/min). RESULTS: Anaemia was observed in 16% of the diabetic patients without CKD; it was more frequent in the diabetic patients with CKD than in the non-diabetic patients with CKD (61.7 vs. 52%, p < 0.05). The comparison among the tertiles showed that the prevalence of anaemia was significantly higher only in diabetic CKD patients of stages 4 and 5. The prevalence was higher in females independently of type 2 diabetes mellitus. In diabetics with a normal renal function, the haemoglobin levels were higher than in diabetics and non-diabetics with CKD, but the diabetics showed lower levels of haemoglobin than non-diabetics at stage 3 and stage 4 of CKD. CONCLUSIONS: Diabetic patients with CKD of stages 4 and 5 have a higher prevalence of anaemia than non-diabetic patients with comparable glomerular filtration rate. A higher awareness of this risk will allow earlier diagnosis and treatment. PMID- 17135771 TI - Long-term outcome of paediatric renal transplantation: follow-up of 300 children from 1973 to 2000. AB - BACKGROUND/AIM: To report our experience of paediatric renal transplantation at Great Ormond Street and Royal Free Hospitals since the inception of the programme. METHODS: Retrospective review of the patient and transplant survival and influencing factors in the 300 children transplanted between 1973 and 2000. RESULTS: 300 children had received a total of 354 transplants; 56 were living related donations. The median age at transplantation was 10.3 (range 1.4-17.9) years. Forty-four percent had congenital structural abnormalities of the urinary tract. Forty-six children required a second and 8 a third transplant before transfer to an adult unit. The overall patient survival at 5, 10, and 20 years was 97, 94, and 72%, respectively. In the overall cohort, the donor type (deceased donor or living-related donor) did not affect mortality, nor did age at transplantation, but those transplanted before 5 years of age had a significantly shorter post-transplant survival time (p < 0.0001). Transplant survival (first transplant) for deceased and living-related donors was 66 and 87% at 5 years (p < 0.01), 51 and 54% at 10 years, and 36% at 20 years (deceased-donor transplants only). Although the overall transplant survival was inferior in children transplanted before 2 years of age (p < 0.03), in the most recent cohort (1990 2000), age did not affect the outcome. On multiple regression analysis, the only predictor of transplant survival was the era of transplantation (p < 0.001). The median final height was within the normal range for males and females; 7 patients received growth hormone after transplantation. CONCLUSIONS: The outlook for successful transplantation is improving, and in the last decade was unaffected by age at transplantation. The survival of living-related donor transplants is superior to deceased-donor transplants for the first 5 years. From the above data, we can predict that a 10-year-old child receiving a renal transplant in 2000 and on ciclosporin-based immunosuppression can expect a transplant half-life of 13.1 years from a living-related donor and one of 10.8 years from a deceased donor transplant. PMID- 17135772 TI - Intraductal papillary mucinous neoplasms and chronic pancreatitis. AB - BACKGROUND: Intraductal papillary mucinous neoplasms (IPMNs) may present with clinical and radiological pictures resembling those of chronic pancreatitis (CP). AIMS: To compare the clinical and epidemiological characteristics of patients suffering from CP with those of patients suffering from IPMN. To assess whether CP is associated with an increased risk of developing IPMN. METHODS: In our departments, from 1981 to 1998, we prospectively followed 473 patients suffering from CP, including 93 cases of chronic obstructive pancreatitis (COP), and 45 patients with a histologically confirmed diagnosis of IPMN. Another 6 patients had an initial diagnosis of CP and a subsequent diagnosis of IPMN. RESULTS: Patients with IPMN were more often female (females 53 vs. 15%; p < 0.001), were older (mean age 63.1 vs. 42.8 years; p < 0.001), drank less alcohol (19 vs. 107 g/day; p < 0.001) and smoked fewer cigarettes (mean 8 vs. 21 cigarettes/day) than CP patients. These results were also confirmed when considering only patients with COP. The 6 patients with a subsequent diagnosis of IPMN were males (p n.s.) with a mean age of 51.4 years (p < 0.05). Only 1 patient was a drinker (p < 0.05) and 4 were smokers (p n.s.). Comparing CP and IPMN, logistic regression analysis selected sex, age, alcohol and smoking, whereas only sex and age were selected when comparing COP vs. IPMN. CONCLUSIONS: In general patients with IPMN present different epidemiological characteristics than those with CP and the subgroup with COP. The clinical and pathological features suggest that in most cases IPMN is the cause of CP and not vice versa. PMID- 17135773 TI - Neuroprotective effects of resveratrol against intracerebroventricular colchicine induced cognitive impairment and oxidative stress in rats. AB - Alzheimer's disease is a complex and multifactorial neurodegenerative disease. Central administration of colchicine, a microtubule-disrupting agent, causes loss of cholinergic neurons and cognitive dysfunction that is associated with excessive free radical generation. The present study was aimed at evaluating the effects of trans-resveratrol in the prevention of colchicine-induced cognitive impairment and oxidative stress in rats. Intracerebroventricular administration of colchicine (15 microg/5 microl) induced impaired cognitive functions in both the Morris water maze task and the elevated plus-maze task. Chronic treatment with resveratrol (10 and 20 mg/kg, p.o.) for a period of 25 days, beginning 4 days prior to colchicine injection, significantly improved the colchicine-induced cognitive impairment. Intracerebroventricular colchicine injection resulted in free radical generation characterized by alterations in oxidative stress markers with a significant increase in malondialdehyde (MDA) and nitrite levels and depletion of reduced glutathione (GSH) activity in the rat brains. It also showed a significant decrease in acetylcholinesterase activity. Besides improving cognitive dysfunction, chronic administration of resveratrol significantly reduced the elevated MDA and nitrite levels and restored the depleted GSH and acetylcholinesterase activity. Results of the present study indicated that trans resveratrol has a neuroprotective role against colchicine-induced cognitive impairment and associated oxidative stress. PMID- 17135774 TI - Angiotensin II-induced MAPK phosphorylation mediated by Ras and/or phospholipase C-dependent phosphorylations but not by protein kinase C phosphorylation in cultured rat vascular smooth muscle cells. AB - Angiotensin II (Ang II) induces a rapid increase in mitogen-activated protein kinase (MAPK) activity through the Ang II type 1 receptor in cultured rat vascular smooth muscle cells (VSMCs). In the present study, we examined the effects of the phospholipase C (PLC) inhibitor U73122, the protein kinase C (PKC) inhibitor GF109203X, and the Ras inhibitor farnesylthiosalicylic acid (FTS) on Ang II-induced activation of p42/p44 MAPKs in cultured VSMCs. Phosphorylation was shown using the Western blot technique with specific phospho-antibodies against MAPK proteins. The PLC inhibitor U73122 abolished the Ang II-induced MAPK activity, while the PKC inhibitor GF109203X only decreased it. There was also an inhibition observed with the Ras inhibitor, FTS on Ang II-induced MAPK activity. These data suggest that Ang II-induced MAPK phosphorylation through the Ang II type 1 receptor could be mediated by Ras and/or PLC-dependent phosphorylations but not by PKC phosphorylation. PMID- 17135776 TI - Gonadotropins and prostate cancer: revisited. AB - Luteinizing hormone and follicle-stimulating hormone are called gonadotropins, because they stimulate the gonads - in males the testes and in females the ovaries. They are not necessary for life, but are essential for reproduction. In addition, the association of these hormones with prostate cancer has been the interest of many researchers. Their detection in the human prostate has been investigated using different methods, including immunologic and RT-PCR techniques. In addition, the increasing evidence of paracrine/autocrine functions of the gonadotropic glycoprotein hormones, their allocation to the superfamily of cystine knot growth factors, and luteinizing hormone/chorionic gonadotropin receptor gene expression in non-gonadal tissues led many researchers to investigate intraprostatic glycoprotein hormones and their receptor gene expression. We aim in this review to shed light on the physiology of the gonadotropins and their association with prostate cancer and highlight the future possibilities of their use as targets in treating this disease. PMID- 17135777 TI - Repositioning and removal of an intra-renal migrated ureteric Memokath stent. AB - The increasing use of ureteric stents including permanent ones is not without their own set of problems. The authors describe the difficulties encountered and a novel technique to reposition a permanent ureteric stent migrated into the kidney with an illustrated case. The same technique can also be applied to a traumatically remove the stent if required. PMID- 17135778 TI - Pitfalls with MRI evaluation of prostate cancer detection: comparison of findings with histopathological assessment of retropubic radical prostatectomy specimens. AB - INTRODUCTION: To assess differences between MRI findings and histopathologically defined prostate cancer localization, we compared clinical results with mapping of radical prostatectomy specimens, and conducted a retrospective MRI cancer localization re-assessment by a urologist-technician after surgery. METHODS: We performed MRI for a total of 37 suspected prostate cancer patients. Subsequently, all underwent retropubic radical prostatectomy after prostate biopsy for confirmation of the diagnosis. All the specimens were studied histopathologically with serial sectioning using a whole organ approach. RESULTS: Of the 37 patients, 26 had positive MRI findings. All the surgical specimens contained cancerous lesions, and 23 had multiple foci. Twenty-four of the MRI-positive cases (92.3%) demonstrated coincidence of both MRI and histopathologically defined lesions. In the single focus cases, 78.6% (11/14) demonstrated exact coincidence, but in the multifocal cases there were no cases with exact coincidence of MRI and histopathological findings (0/23). CONCLUSION: MRI evaluation cannot be considered an effective diagnostic tool in itself for detection of prostate cancers because sensitivity is far from satisfactory, especially in multi-focal cases. PMID- 17135779 TI - Should prostate cancer status be determined in patients undergoing radical cystoprostatectomy? AB - INTRODUCTION: We estimate the frequency of prostate cancers detected incidentally in radical cystoprostatectomy specimens and discuss whether the prostate cancer status should be determined in patients undergoing radical cystoprostatectomy. MATERIALS AND METHODS: A total of 97 radical cystoprostatectomies without evidence of prostate cancer on digital rectal examination were performed for transitional cell carcinomas of the bladder between January 2001 and May 2004. The mean patient age at the time of surgery was 66.9 +/- 9.52 (range 49-75) years. RESULTS: The overall incidence of prostate cancer detected in radical cystoprostatectomy specimens was 21.6% (21/97 specimens). The mean tumor volume was found to be 0.93 +/- 0.81 ml. The tumor volume was >0.5 ml in 12 cases (57.1%). The surgical margin was negative in all cases, and the disease was organ confined in 20 patients (95.2%). Capsular invasion was evident in 2 patients (9.5%), 1 of whom had lymph-node-positive disease. CONCLUSIONS: Despite the high prevalence of incidental prostate carcinomas among patients with bladder cancer undergoing cystoprostatectomy, the vast majority of the cancers are organ confined. However, the prostate cancer status should be determined on the basis of digital rectal examination and prostate-specific antigen in patients undergoing radical cystoprostatectomy - especially if prostate-sparing cystectomy is planned. PMID- 17135780 TI - Prescription practice for LH-RH analogs and androgen receptor antagonists in the treatment of advanced prostate cancer in Germany (1993-2003). AB - INTRODUCTION: Nearly 80% of the patients suffering from advanced stages of prostate cancer in Germany are treated currently with luteinizing hormone releasing hormone (LH-RH) analogs alone or in combination with androgen receptor antagonists. The aim of this study was to assess whether the prescription practice for LH-RH analogs and direct antiandrogens between 1993 and 2003 parallels the rise in the incidence rates for prostate cancer associated with the proportional rise in daily doses. METHODS: Between 1993 and 2000 a yearly randomly selected number of receipts has been evaluated from official pharmacies in Germany which have been billed to public health insurance companies. Data concerning drug use, number of daily doses, and total cost of the respective drugs have been calculated by the Research Institute of the AOK (Wissenschaftliches Institut der AOK, WIdO). These prescription data have been related to incidence and mortality rates of patients with prostate cancer. RESULTS: Between 1993 and 2003 the age-standardized incidence rate of carcinomas of the prostate has risen by 180%, while the disease-specific mortality practically has remained nearly at the same level. The daily prescribed doses of LH-RH analogs have risen by a factor of 8.5 from 3.9 to 33.5 million daily doses during this time. In contrast, the daily doses of androgen receptor antagonists have remained unchanged between 1997 and 2001, followed by a rise of 50% between 2001 and 2003. CONCLUSIONS: The first analysis of the prescription practice of antiandrogens in the treatment of prostate cancer with respect to the incidence rates in Germany could only be facilitated by the fact that these medications are in exclusive use for this indication in Germany. The rise in prescriptions may be explained by the therapy shift from surgery to LH-RH application until the year 2000; the reasons for a further rise since 2000 remain speculative. In conclusion, only prevalence data would allow the exact calculation of medication needs among prostate cancer patients. PMID- 17135781 TI - Spontaneous knotting of urinary catheters: clinical and experimental observations. AB - Urinary catheters are known to spontaneously knot inside the bladder. Approximately 40 cases of this complication have been recorded in the world literature. Clinical observation in 4 children and simulation experiments done using a balloon model are described herein. Risk factors identified in this study includes catheter size less than 10 Fr, overdistended bladder and insertion of more than 10 cm length of catheter inside the bladder. Water-current generated by the flow of urine appears to play an important role in the pathogenesis of catheter knotting. PMID- 17135782 TI - Voiding pattern of healthy Taiwanese women. AB - AIM: Few studies have examined the voiding pattern of healthy females, despite the importance of such research for diagnosing abnormal voiding behavior. This study investigates the voiding behavior of healthy women in Taiwan. METHODS: 68 healthy women, aged 19-66 years and claiming to void normally were enrolled. Twenty-two of the subjects (32.4%) were postmenopausal. Each woman completed a 3 day voiding diary. The diary recorded urine volume, voiding frequency, urine volume per voiding for the whole day (24 h), and urine volume during both daytime and nighttime. Age and various other voiding parameters were correlated, and the voiding patterns were compared between premenopausal and postmenopausal women. RESULTS: All women voided 7.34 +/- 1.63 times daily. Higher age was associated with higher whole day and daytime voiding frequency. Nocturia was not common for the group as a whole, with a mean nocturnal voiding of 0.25 times per night. However, higher age was significantly associated with a higher nighttime voiding frequency. A significantly positive correlation was also observed between age and whole day urine volume. Compared with regularly menstruating women, postmenopausal women had a significantly higher whole day and nighttime voiding frequency. Menopause and age both independently affect voiding parameters after multivariate analysis. CONCLUSIONS: The results of this study provide invaluable baseline data on female voiding behavior. Additionally, this study found that for healthy women, age and menstruation status significantly affect voiding patterns. Analysis of female voiding behavior thus should consider these two factors. PMID- 17135783 TI - Overactive bladder syndrome among community-dwelling adults in Taiwan: prevalence, correlates, perception, and treatment seeking. AB - PURPOSE: We investigated the prevalence, correlates, perception of overactive bladder syndrome (OAB) and doctor-consultation behavior among community adults. SUBJECTS AND METHODS: This urological survey was part of a government-supported health utilization project conducted in Matsu, Taiwan. Participants (n = 1,827) completed a questionnaire collecting information on OAB, which was defined as urgency with either frequency or nocturia. RESULTS: The age-adjusted prevalence of OAB was 16.9%, including 4.5% with urge incontinence. Age (p = 0.002), diabetes (p = 0.03), and benign prostatic hyperplasia (p < 0.001) in men, whereas diabetes (p = 0.01), hyperlipidemia (p = 0.03), stress incontinence (p < 0.001), and recurrent lower urinary tract infections (p = 0.02) in women were factors independently associated with OAB. Most people with OAB (69.0%) perceived little bother; only 13.0% had consulted doctors for this problem. CONCLUSION: Symptoms related to OAB are common among community-dwelling adults, yet most perceive it as a minor problem. Our results identify several factors that are significantly associated with OAB. PMID- 17135784 TI - Propiverine hydrochloride immediate and extended release: comparison of efficacy and tolerability in patients with overactive bladder. AB - INTRODUCTION: This study aims to compare the efficacy of propiverine hydrochloride immediate release (IR), propiverine hydrochloride extended release (ER) and placebo for the treatment of overactive bladder syndrome. The primary outcome measure is incontinence episode frequency, with secondary outcome measures including mean volume per void and quality of life as assessed on King's Health Questionnaire. MATERIAL AND METHODS: The double-blind, double-dummy, randomized study compared IR 15 mg twice daily, ER 30 mg once daily and placebo in 3 parallel groups. After a run-in period of 7 days, the patients were treated for 32 days. Nine hundred and eighty-eight patients were randomized, and 910 patients completed the protocol without major violations. RESULTS: The number of incontinence episodes/24 h decreased by 2.26 in the IR group (p < 0.001 vs. placebo), by 2.46 in the ER group (p < 0.0001 vs. placebo) and by 1.75 in the placebo group. The most frequent adverse event was dry mouth with 22.8% of the patients in the IR group, 21.7% in the ER group and 6.4% in the placebo group. The overall tolerability was rated 'very good' or 'good' by more than 80% of the investigators and patients in all 3 groups. CONCLUSIONS: Propiverine ER 30 mg once daily and propiverine IR 15 mg twice daily significantly reduce the number of incontinence episodes/24 h within a treatment period of 32 days. Both formulations are safe and well tolerated. The extended release formulation of propiverine is a suitable new option for the treatment of the overactive bladder. PMID- 17135785 TI - Impact of percutaneous access point number and location on complication and success rates in percutaneous nephrolithotomy. AB - OBJECTIVES: Percutaneous nephrolithotomy (PNL) is sometimes associated with complications, especially in patients with complex stones. Herein, we review our experience with PNL to determine the impact of percutaneous access number and location on success and complication rates. PATIENTS AND METHODS: During a 2-year period, a total of 275 patients with a mean age of 42.3 +/- 14.8 (range: 13-75) years underwent PNL. Stones were classified as simple in 51.6%, and complex (staghorn calculi or renal pelvis stones coexisting with caliceal stones) in 48.4%. Percutaneous access was done under C-armed fluoroscopy and the tract was formed with a high-pressure balloon dilation system. One single percutaneous access was sufficient in 210 (76.4%), while 2 accesses were utilized in 44 (16%), and > or =3 accesses in 21 cases (7.6%). Supracostal access was performed in 23 (8.4%) patients. RESULTS: An overall success rate of 94.9% was achieved. Stone location, but not the access point location, was the major determinant for success, which was 99.3 and 90.2% in patients with simple and complex stones, respectively (p < 0.01). Significant complications included bleeding necessitating blood transfusion in 28 (10.2%), and hydropneumothorax in 2 (0.7%) patients. Bleeding was observed in 39.1 and 7.5% of patients managed with supracostal access, and subcostal access, respectively (p < 0.01). An increased number of access points significantly augmented the risk for bleeding. Bleeding was encountered in 7.6% of patients managed with 1 percutaneous access point, and in 18.5% of cases managed with > or =2 access points (p < 0.05). Hydropneumothorax occurred in patients with supracostal access. CONCLUSION: Supracostal access as well as multiple punctures may be needed especially in the management of complex stones, and the need for multiple access points and supracostal access significantly increases complication rates. PMID- 17135786 TI - A comparative study of protein profiling by proteomic analysis in camptothecin resistant PC3 and camptothecin-sensitive LNCaP human prostate cancer cells. AB - INTRODUCTION: Drug resistance is a major obstacle for the therapy of prostate cancer, but its underlying mechanisms are not clarified. To detect some candidate marker proteins which may confer resistance to the anticancer drug camptothecin (CPT; DNA topoisomerase 1 inhibitor), the current study deals with the comparative proteomic profiling of CPT-resistant PC3 and CPT-sensitive LNCaP human prostate cancer cell lines which have been widely employed as a useful model to investigate prostate cancer cells. MATERIALS AND METHODS: The global profiling of the protein expression was investigated in CPT-resistant PC3 and CPT sensitive LNCaP prostate cancer cells using 2-dimensional polyacrylamide gel electrophoresis/matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. RESULTS: 144 proteins were identified and their expression levels were compared between the two cell lines. Four proteins - annexin A1, glutathione S-transferase (GST) pi, galectin (Gal) 3 and glucose-regulated protein 78/Bip - that are suggested to contribute to the development of drug resistance were found to be preferentially or highly expressed in PC3 cells, whereas LNCaP cells did not show detectable expression of annexin A1, GST-pi and Gal-3. CONCLUSION: The expression level of these proteins and/or mRNAs could be a useful parameter to evaluate the chemotherapy resistance in clinical specimens of prostate cancer. PMID- 17135787 TI - Additional gene therapy with rAAV-wt-p53 enhanced the efficacy of cisplatin in human bladder cancer cells. AB - OBJECTIVE: Gene therapy is defined as the treatment of an acquired or inherited disease by transfer of genetic material. The most common strategies in gene therapy of bladder cancer are corrective, inductive and cytotoxic gene therapy. Mutations in the p53 tumor suppressor gene are the most common molecular genetic abnormalities in bladder cancer and p53 gene transfer in the human bladder cancer cell line by adenoviral or other vectors was demonstrated to be cytotoxic. However, so far there has been no report of adeno-associated virus-2 vector mediated p53 gene deliveries in bladder cancer. In this study, wild-type p53 cDNA was transfected into the bladder cancer cells, using the adeno-associated virus-2 vector, and the capability of rAAV-wt-p53 gene therapy in bladder cancer was evaluated in vitro. METHOD: Bladder cancer cell lines 5637 were transduced with adeno-associated virus-2 vectors containing wild-type human p53 gene (rAAV-wt p53). Gene expression and transcriptional activation of p53 was determined by Western blot analysis. The cellular growth inhibition and apoptosis of rAAV mediated p53 transfection were assessed by flow cytometry. The combination effect of rAAV-wt-p53 and cisplatin was measured by MTT assay. RESULTS: The virus rAAV efficiently enters the cells and expresses its gene products. The gene product of rAAV-wt-p53 is cytotoxic to bladder cancer cells. The bladder cell line 5637 was found to experience a synergistic killing effect when rAAV-wt-p53 was used in combination with cisplatin. CONCLUSION: rAAV-mediated p53 gene transfer could offer a powerful novel therapeutic approach in bladder cancer. PMID- 17135788 TI - Endourological re-establishment of a disrupted infundibulum after renal laceration. AB - We report a case of severe urinary extravasation after renal contusion and its successful management by endoscopic creation of a neoinfundibulum. When the stenotic infundibulum cannot be traversed with a guide wire, creation of a new infundibulum will offer a secure alternative for accessing the collecting system. PMID- 17135789 TI - Recurrent arteriovenous malformation of the scrotum secondary to pelvic trauma. AB - We describe a case of recurrent scrotal arteriovenous malformation (AVM) in a young man secondary to pelvic fracture sustained during a road traffic accident. This was diagnosed as a lipoma on the ultrasound and clinical examination. To our knowledge, this is the first reported case of recurrent scrotal AVM secondary to trauma. PMID- 17135790 TI - Giant urethral diverticulum: unusual complication following ventral buccal mucosa onlay graft urethroplasty. AB - During the past 10 years, buccal mucosa grafting for urethroplasty has gained widespread acceptance. In general the reported success rates are 80% and higher. Typical complications are recurrent strictures, fistula, and, especially when the technique of ventral onlay urethroplasty has been used, sacculations that are of little clinical significance in most cases. The development of large diverticula has not been described yet. We report on the case of a 63-year-old man who developed a giant diverticulum of the urethra 1 year after a ventral buccal mucosa onlay graft urethroplasty for recurrent bulbar urethral strictures had been worked out. PMID- 17135791 TI - Isolated unilocular renal hydatid cyst: a rare diagnostic difficulty with simple cyst. AB - Hydatid disease is endemic in many parts of the world. The location is mostly hepatic (75%) and pulmonary (15%), and only 10% of the cysts occur in the rest of the body. It may develop in almost any part of the body. Although hydatidosis can be present in all parts of the human body, urinary tract involvement develops in only 2-4% of all cases, and isolated renal cysts are extremely rare. There are no specific clinical symptoms or signs that will reliably confirm the diagnosis of renal echinococcosis. In addition, there is no laboratory finding that is pathognomonic for hydatid disease except for hydatiduria. Routine blood tests are generally normal except for eosinophilia which is found in only 50% of the cases. Radiological studies have a more important place in the preoperative diagnosis of renal hydatic disease. However, there is no specific sign on plain radiography or intravenous urography, and ultrasound or computed tomography cannot always show a hydatidosis as a specific lesion. From these reasons, sometimes it is difficult to differentiate between a unilocular hydatid cyst without mural calcification and a simple renal cyst. So, despite its rarity, hydatid disease should be included in the differential diagnosis of cystic lesions in solid organs or other anatomic sites, especially in endemic countries. We present herein a patient with isolated renal hydatid cyst mimicking a simple cyst treated successfully with complete excision. PMID- 17135792 TI - Re: using two ipsilateral double J ureteral stents for extrinsic ureteral obstruction due to colon carcinoma. PMID- 17135793 TI - Understanding adolescent depression in ethnocultural context: updated with empirical findings. AB - As a continuation of the discussion of the ethnocultural variations in adolescent depression, a cross-sectional, school-based study was designed to examine ethnocultural variations in depression and somatic symptoms in white, African American, Hispanic American, and Asian American adolescents (N = 307). Ethnocultural variations in the relationship between depression and somatic symptoms were also explored. Each ethnocultural group of adolescents had its own particular tendency to symptoms of depression. Ethnocultural-specific patterns of relationship were also found between depression and somatic symptoms. This study confirmed the importance of understanding ethnocultural variations in adolescent depression. PMID- 17135794 TI - Development of a social justice gauge and its use to review the Canadian Nurses Association's Code of Ethics for Registered Nurses. AB - Betty Bekemeier and Patricia Butterfield undertook a critical review of 3 American nursing documents in relation to the concept of social justice. Their article inspired a review of the Canadian Code of Ethics for Registered Nurses, using a Social Justice Gauge developed by the Canadian Nurses Association. The article outlines the development of the gauge and its use in this review. Although some evidence of generic and outdated language is evident in the Canadian code, the text appears well aligned with social justice ideals overall. That being said however, there still remains significant possibility for enlarging the application of social justice, especially in relation to the place of nurses in healthcare institutions and in nontraditional nursing settings, in future revisions of the code. Work to further examine, adapt, and test the Canadian Nurses Association's Social Justice Gauge is encouraged. PMID- 17135797 TI - Holism and physiology: continuing the discussion. PMID- 17135798 TI - Hall's authentic meaning of medicalization: an extended discourse. AB - The purpose of this article is to inform emerging theoretical understanding of medicalization by challenging, updating, and affirming a previously published essay on this topic by Hall. We accomplished this through the creation of a discourse among the 3 authors using the essay, written entirely from a personal perspective, and integrated our own personal experiences as nurses and patients. Hall's essay on the authentic meaning of medicalization provided a critical understanding of medicalization of cancer describing the factors, forces, and consequences, seeking to raise consciousness and provoke reform. The 3 of us added our voices to her narrative seeking to expand the discourse on medicalization and inform theory development. We have shared our individual and collective voices and identified elements that might point the way to theoretical emergence and ending with our own call for nursing to evolve further as a field worthy of human inspiration. PMID- 17135800 TI - The nurse-patient relationship in cancer care as a shared covenant: a concept analysis. AB - The concept of the nurse-patient covenant was explored using a multiple-method research design. Analysis of the literature (167 references) was the basis for 2 focus group interviews (12 nurses) and 8 semistructured patient interviews. Attributes, antecedents, and consequences were identified through an analysis of findings across all data sources. The nurse-patient covenant in cancer care is defined as an enduring relationship, embodying caring benevolence and contextually negotiated reciprocity. It is preceded by the demonstration of nursing competence as a manifestation of nursing tradition and leads to patients' trusting and entrusting nurses, the emergence of personhood and professionhood, and benefits and obligations. PMID- 17135801 TI - Toward a critical theoretical interpretation of social justice discourses in nursing. AB - Despite widespread appeals to social justice, nursing conceptions of this ideal have been critiqued as incomplete and inconsistent. With the aim of contributing to a critical dialogue on discourses of social justice in nursing, we explore contemporary theories of social justice and their move beyond a distributive paradigm, employing techniques of replication and critique of social justice discourses in nursing. We consider how postcolonial feminist theory can help us understand the relevance of more recent critical interpretations of social justice, particularly in reinterpreting and broadening nursing's individualistic focus on social justice so that due consideration and actions are directed toward the intersecting impact of historically and socially mediated conditions on health and human suffering. PMID- 17135802 TI - A middle range theory for generative quality of life for the elderly. AB - Decades of quality of life (QOL) research has failed to produce widely recognized QOL definitions or measurements. The inconsistency may be due to an ontological error in the perspective of researchers. Most researchers portray QOL as a given or inherent condition that declines in the face of challenges. We believe QOL is a cumulative process that results from a series of connections and disconnections that elders experience in their daily lives. The Register theory of Generative Quality of Life for the Elderly offers an alternative ontological perspective by placing elder QOL in a generative context. PMID- 17135803 TI - Teen mothers and their teenaged children: the reciprocity of developmental trajectories. AB - This article reports on the fifth wave of a multigenerational study of teen mothering. Two paradigm cases reveal how teen mothers' agency and development are linked with their teenaged children's development in a world shaped by poverty and limited resources. Teens with attenuated relationships and without anchors of hope and behavioral expectations for the future face an existential void that sets them adrift. Teens with well-developed parental relationships, family routines, and behavioral expectations have a stake in the future. This emic view shows how teens respond to the challenges of adolescence shaped by mothers' agency, family practices, class, and culture. PMID- 17135804 TI - Supplement editorial: alzheimer disease. PMID- 17135805 TI - ADCS Prevention Instrument Project: overview and initial results. AB - One objective of the Alzheimer's Disease Cooperative Study (ADCS) is to develop new or improved instruments and assessment methods for evaluating treatment efficacy in Alzheimer disease (AD) clinical trials. The ADCS Instrument Committee has previously helped to define the state of the art in assessment for AD and Mild Cognitive Impairment clinical trials. We are now entering an exciting era of primary prevention trials to evaluate promising treatments that may delay disease onset and there is a need to develop appropriate instruments for these trials. The ADCS instrument committee has undertaken a project to develop instruments for prevention studies that assess domains known to be important in AD. Prevention trials are long and require large numbers of subjects, making them costly and requiring a high burden of participation for subjects. The current study focused on developing instruments that can be completed at home and in the clinic. The instruments are being evaluated in a cohort of nondemented elderly participating in a 4-year longitudinal study that simulates the design of a primary prevention trial. This report describes the design, baseline characteristics, and some longitudinal outcomes of the study cohort through the completion of the first 2 years of follow-up. We also describe the assessment domains to be measured with our new experimental instruments. This study recruited 644 subjects, 75 years of age and older. Participation in a "book club" that provided free books of interest to elders was offered as a recruitment incentive. Approximately 23% had some mild cognitive symptoms consistent with a Clinical Dementia Rating of 0.5. All subjects received a standardized in-clinic evaluation at baseline, which is repeated annually for 4 years to identify cases suspected of developing dementia and to measure longitudinal change on established clinical assessments. Subjects completed a set of self-administered experimental instruments at home or in the clinic designed to assess cognitive function and behavior, global change, activities of daily living, quality of life, and resource use. An additional "mail-in cognitive function questionnaire" was obtained separately by mail, 1 month before the other assessments. To evaluate the feasibility, efficiency, and validity of the home-based instruments in comparison with acquiring the same information during a clinic visit, subjects were randomized to 1 of 2 conditions in which the baseline and annual follow-up assessments are completed either at home ("home group") or at the study site during their clinic visits ("clinic group"). This initial report describes the ongoing 4-year longitudinal study and provides baseline results, which confirm the feasibility of obtaining home-based clinical information via mail or telephone. Initial results for the experimental instruments and for the book club are reported in separate accompanying articles. PMID- 17135806 TI - ADCS Prevention Instrument Project: ADCS-clinicians' global impression of change scales (ADCS-CGIC), self-rated and study partner-rated versions. AB - BACKGROUND: Because primary prevention trials will require large samples and modest treatment effects are expected, the use of standard clinician administered, clinic-based measures are unlikely to be feasible. There is a need for proxy-administered outcome measures. The goal of the Alzheimer's Disease Cooperative Study (ADCS) Prevention Instrument Project was to conduct a simulated Alzheimer disease prevention trial in 650 nondemented elderly (Ferris et al, 2006). This involved comparison of data acquisition from both home and clinic and the use of both informant-ratings and self-ratings. Important outcomes included clinical global impressions of change (CGIC) as indicators of clinically meaningful change. Such ratings provide verification that the effects of a medication as measured on rating scales are readily observable and clinically meaningful. One objective was to develop self-rated and study partner-rated CGICs optimized for nondemented elderly or people with very early Alzheimer disease. An important consideration was whether global assessments are specific and sensitive measures of change during a prevention trial. METHODS: A self-administered CGIC and a study partner-rated CGIC were developed to be used either in the clinic or at home. Using 3-month follow-up data, we determined its reliability and validity with 317 subject-partner pairs. We compared subject-ratings with partner-ratings, clinic-based with home-based ratings, and ratings based on severity as determined by the Clinical Dementia Rating scale. RESULTS: There were no differences between clinic and home ratings. Overall, 24% of subjects rated themselves, and 10% of study partners rated the subjects, as minimally to markedly improved. Subjects and partners agreed to within 1 point of their ratings 83% of the time on the 7 point scale. There were weak correlations, generally <0.20, with change scores of selected clinical rating scales. DISCUSSION: The CGICs behaved as expected, showing no overall change over 3 months, no difference between administrations at home compared with clinics, and concurrent validity. Some subjects tended to rate themselves better than their partners rated them. These analyses show the potential for using home-based CGICs which can be completed with minimal supervision and allow assessments of potential preventative interventions. PMID- 17135807 TI - ADCS Prevention Instrument Project: development of a brief verbal memory test for primary prevention clinical trials. AB - The validity and reliability of clinic-based and telephone-based versions of a 4 word delayed recall test were evaluated in nondemented elderly individuals (n=644) participating in a simulated primary prevention clinical trial. There was no significant difference in the average scores achieved by participants tested in clinic (mean=3.40) or by telephone (mean=3.47) and the 2 groups had similar distributions of scores. Delayed recall scores were significantly, but weakly, correlated with scores on a rigorous verbal memory task, were lower in participants in Clinical Dementia Rating stage 0.5 than in those in Clinical Dementia Rating stage 0, and were lower in those with subjective memory complaints than in those without complaints. There was only fair correspondence between scores achieved at initial testing and 3 months later for both versions of the test. There were no differences in the average scores achieved by men or women, those older (age 80 to 93) or younger (age 75 to 79) than age 80, or those with white or nonwhite ethnicity. Participants with low education scored significantly lower than those with high education. Results suggest that clinic based and telephone-based versions of the Four Word Delayed Recall Test are valid and reliable and can be used to screen for possible memory deficits in elderly individuals. However, the psychometric properties of the test are relatively weak and do not support the general use of the test for clinical and research purposes if the use of a more rigorous memory test with a wider range of possible scores is feasible. PMID- 17135808 TI - ADCS Prevention Instrument Project: behavioral measures in primary prevention trials. AB - BACKGROUND: There is an urgent need for the development of inexpensive, reliable, and valid instruments that can be used in large-scale primary prevention trials of compounds aimed at ameliorating progression from normal aging to mild cognitive impairment or Alzheimer disease. The Alzheimer's Disease Cooperative Study launched a Prevention Instrument Project to develop such methodologies. Behavioral changes are common in diseases causing dementia and may occur prior to a point when cognitive changes are sufficiently severe to allow diagnosis of a dementia syndrome. Experimental behavioral measures were included in the protocol to examine this hypothesis. METHODS: Six hundred forty-four individuals with CDR 0 or 0.5 were randomly assigned to receive a brief in-clinic behavioral assessment or telephonic administration of the same assessment. The questions were asked to the individual and their research partner. The Prevention Instrument Project included behavioral measures of depression, anxiety, irritability, and apathy. RESULTS: All measures demonstrated acceptable test retest reliability at 3-month intervals except for the single-item depression screen by the subjects' research partner. Behavioral changes are significantly more common among patients with Clinical Dementia Rating (CDR) scores of 0.5 compared with CDR scores of 0. Behavioral alterations including irritability, anxiety, and apathy are more common among ethnic minorities than among the White population. Depression, irritability, anxiety, and apathy are significantly correlated with each other. CONCLUSIONS: Behavioral changes are common among those with mild degrees of cognitive compromise (CDR 0.5). Telephonic assessment of behavioral changes is feasible. The predictive value of these alterations for progression to Alzheimer disease or other dementias will be assessed longitudinally. PMID- 17135809 TI - ADCS Prevention Instrument Project: assessment of instrumental activities of daily living for community-dwelling elderly individuals in dementia prevention clinical trials. AB - BACKGROUND: In primary prevention trials for Alzheimer disease, the inception cohort typically has normal or minimally impaired complex activities of daily living (ADL). ADL change during a trial could trigger detailed evaluation or serve as an outcome measure. A brief, easily administered, and reliable ADL rating scale would assist prevention studies. OBJECTIVES: To develop an ADL scale for prevention trials that allows self-rating or completion by informants. METHODS: The Activities of Daily Living-Prevention Instrument (ADL-PI) was developed, comprising 15 ADL and 5 physical function questions. Six hundred forty four elderly subjects participating in the Prevention Instrument Project completed a self-rated version of the ADL-PI, and informants for 632 subjects completed an informant version. Informants also completed a Mild Cognitive Impairment (MCI) ADL questionnaire to allow comparisons. RESULTS: Subjects performed well on all ADL scales at baseline. Completion of the ADL-PI questionnaires at home or in-clinic yielded comparable information. Scores from baseline to 3 months had good reliability. The ADL-PI, obtained from either self report or informants, discriminated between subjects rated as CDR 0 and CDR 0.5. Subjects with worse baseline cognitive performance also had slightly worse ADL-PI scores. Preliminary analysis indicates that subjects who triggered cognitive evaluations had slightly lower baseline ADL-PI scores by both self and informant reports. CONCLUSIONS: The ADL-PI can be completed at home or in clinic, and has adequate reliability. The utility of self-administered and informant versions and predictive value of reported deficits requires further follow-up. PMID- 17135810 TI - ADCS Prevention Instrument Project: the Mail-In Cognitive Function Screening Instrument (MCFSI). AB - In future research on the prevention of Alzheimer Disease (AD), a large simple prevention trial might test a safe intervention (eg, vitamins) in a very large population (eg, 10,000 subjects), powered to detect a modest treatment effect size. In a large, simple design, regular assessments must be very low cost, for example, handled by mail. The Mail-In Cognitive Function Screening Instrument (MCFSI) was developed for the Alzheimer's Disease Cooperative Study Prevention Instrument Project to evaluate whether a brief mail-in screening tool can be used as a specific and sensitive trigger for a diagnostic evaluation in the course of a prevention trial. The MCFSI consists of 2 similar sets of 14 questions mailed separately to the subject and the study partner, who are asked to complete them independently. The questions are derived from a standard clinical assessment of dementia; they ask about decline in function over the last year. We are currently evaluating the utility of the instrument in 640 subjects over the course of the "simulated" AD prevention trial. Analysis of baseline data showed significant differences in mean MCFSI scores between sex and ethnic groups (for subjects), age groups (for partners), and Clinical Dementia Rating global score groups (both subject and partner). In this nondemented population, both subject's and partner's MCFSI responses were related to cognitive performance. There was no significant association between MCFSI score for either subjects or partners and apolipoprotein E genotype. The MCFSI captures information related to cognitive and functional status in nondemented elderly individuals; the Alzheimer's Disease Cooperative Study Prevention Instrument Project will determine whether this brief, mail-in questionnaire is useful as a trigger for diagnostic evaluation in an AD primary prevention trial. PMID- 17135811 TI - ADCS Prevention Instrument Project: quality of life assessment (QOL). AB - Information about quality of life (QOL) is valuable in evaluating pharmaceutical agents but it is not adequately assessed in many dementia drug trials. In prevention trials, following participants to conversion to AD requires QOL scales appropriate for both normal and cognitively impaired individuals. Our objective was to evaluate the utility of several scales for subject or informant QOL assessment: Quality of Life-AD; Quality of Life Activity Inventory; SF-36; SF-12 (a shortened version of the SF-36); and Satisfaction with Life Scale. Measurements were collected from 644 subject-study partner pairs, half of whom completed the instruments at the clinic and half at home. Three-month test-retest data were collected. Scales administered at home or in clinic did not differ significantly. Subject self-ratings showed a wide range for all scales. Test retest intraclass coefficients ranged from 0.67 to 0.77. Moderately high interscale associations suggest that the scales are measuring common aspects of QOL but are not equivalent. Furthermore, they differed with respect to associations with demographic variables and QOL determinants. We conclude that the QOL scores at baseline show sufficient range and reliability to suggest they will have utility in tracking QOL through conversion to dementia. PMID- 17135812 TI - ADCS Prevention Instrument Project: pharmacoeconomics: assessing health-related resource use among healthy elderly. AB - BACKGROUND: The Prevention Instrument project of the Alzheimer's Disease Cooperative Study (ADCS) seeks to develop instruments to assess treatment efficacy including potential economic benefit. The Resource Use Inventory (RUI) is an instrument that has been used to capture resource utilization and costs in populations with Alzheimer disease (AD). However, resource utilization and costs for healthy, cognitively intact elderly as they begin to demonstrate cognitive deterioration are not well understood. In addition, the loss that relates to the subjects' own time as they transition through cognitive impairment is not well documented. OBJECTIVES: To evaluate the utility of the RUI in a sample of cognitively intact elderly individuals living in the community and enrolled in AD prevention trials. METHODS: The RUI was administered to 644 subjects and their study partners either at home or in the clinic. For half of each sample, 3-month retesting was carried out. The RUI consisted of 9 questions. The first part of the RUI captured subjects' use of direct medical care (eg, hospitalizations) and nonmedical care (eg, home health aides). The second part of the RUI captured the time caregivers spend providing care to the subjects. The third part of the RUI captured subjects' participation in volunteer work and employment. The assessment interval for each question was the past 3 months. RESULTS: The percentage of RUI forms returned incomplete or inaccurate for both in-clinic and at-home groups was extremely low. There were no differences in utilization rates between in-clinic and at-home group for all items in the RUI. Except for use of outpatient procedures, tests, or treatments, there were no differences in utilization rates between subjects who filled out the RUI with the help of their study partners or by themselves. Items in the RUI were sensitive to subjects' cognitive and functional status and demographic characteristics. CONCLUSIONS: Home-based completion of the RUI by participants in an AD prevention study is feasible, and seems to provide data that are reliable and valid. The instrument will be useful for tracking resource and time use through transition from healthy to cognitive impairment. PMID- 17135813 TI - ADCS Prevention Instrument Project: pilot testing of a book club as a psychosocial intervention and recruitment and retention strategy. AB - Both psychosocial and biologic interventions may delay or prevent Alzheimer disease. Staying mentally active may help older people maintain their cognitive abilities. In the Alzheimer Disease Cooperative Study Prevention Instrument Project a book club was introduced as a recruitment and retention device. A 3-arm study was designed and included: a nonrandomized, self-selected group (n=211) who chose not to participate in the book club, and 2 groups randomly assigned to receive 2 books per year in individual self-improvement (n=210) or community involvement (n=207) categories. Participants reported their reactions to the selections and other reading behaviors. Results from the first 2 years revealed that most book club participants agreed with Likert-type statements indicating the readings were enjoyable (P<0.001), had an impact on their thinking (P=0.01), and were shared by them with others (P=0.002). Respondents in the community involvement group agreed more strongly with these statements than those in the self-improvement category. Comments from participants in response to open-ended questions in the reader survey revealed such themes as developing plans for successful aging and reflecting on attitudes and behaviors in their own lives. Further longitudinal analyses are planned to determine whether the book club influenced retention and whether participation was associated with slowing cognitive decline. PMID- 17135814 TI - Needs of lay caregivers of bone marrow transplant patients in Turkey: a multicenter study. AB - The purpose of this research was to identify the needs of lay caregivers of bone marrow transplant (BMT) patients throughout the BMT trajectory. The sample consisted of 58 lay caregivers of BMT patients (5 of which were inpatients) from the 4 BMT units in Ankara, Turkey. Data were collected through a demographic data form and the Psychological and Social Needs Scale. Data analysis was performed using SPSS 10.0 for Windows. Among the descriptive statistics, frequencies and percentages were used to define the characteristics of lay caregivers and responses related to information. Mean values, standard deviations, and the Pearson correlation coefficients were also calculated for the results of the subscales. Most of the lay caregivers were spouses. The rest were other family members: brother/sister, mother, son/daughter, or father. Only one lay caregiver was a friend. The most common type of BMT was autologous. Most of the surveys were completed by the lay caregivers whose patients had been out of the hospital for 100+ days after BMT. The lay caregivers were educated on the BMT process by various sources, and 67.24% of them expressed satisfaction with the information that they were given. A majority of them preferred face-to-face communication with a healthcare professional. The categories of the information provided were diagnosis and treatment, homecare after discharge, follow-ups and laboratory tests, and nutrition. They indicated a need for more information, particularly regarding homecare after discharge and diagnosis. Fear was the highest scoring psychological needs/problems of lay caregivers, whereas leisure activity deficit was highest among the social needs/problems. The information gathered from this study led us to reevaluate our healthcare services for both patients and lay caregivers to improve physical, psychological, and social aspects of the nursing care as a whole. PMID- 17135815 TI - A systematic review of worldwide cancer nursing research: 1994 to 2003. AB - The aim of this study was to assess the cancer nursing research papers published in the past decade; identify their characteristics in terms of country of origin, participants, settings, diagnostic foci, and methodologic choices; and evaluate their quality. A systematic review was carried out of all published papers in the Cumulative Index of Nursing and Allied Health Literature between the years 1994 and 2003, using the keywords "cancer," "nursing," and "research." A total of 619 papers met inclusion criteria and were evaluated by 5 researchers. Almost half the papers were derived from the United States (49.1%), followed by the UK, Sweden, Canada, and Australia. In more than half of the published papers (52.2%), health professionals (mostly nurses) were the studies' participants. Also, much of the published research used patients with mixed diagnosis, or patients with breast or hematologic cancers. Two-thirds of the studies were quantitative, whereas most studies were descriptive in nature. The quality of both quantitative and qualitative studies was low, with only a small percentage meeting the highest quality criteria. Studies reporting funding and those published in journals with an impact factor showed a higher quality score than those not reporting funding or not published in journals with an impact factor. Cancer nursing research is still in a developmental stage, although it has made a considerable contribution to the evidence base of the discipline. A number of issues need to be tackled before we improve our output, such as organizational or workforce issues, infrastructure support, funding, and methodologic challenges. PMID- 17135816 TI - White cancer patients' perception of gender and ethnic differences in pain experience. AB - Not considering cancer patients' own views and experience with pain, especially gender and ethnic differences in their cancer pain experience, was reported to be a major contributor to the miscommunication that frequently results in inadequate cancer pain management. The purpose of this study was to explore white cancer patients' perception of gender and ethnic differences in pain experience through an online forum. This was a descriptive qualitative study among 29 white cancer patients based on a feminist approach. Nine topics related to cancer pain experience were used. The data were analyzed using thematic analysis: 5 themes were identified. First, the participants perceived that pain accompanies cancer throughout the diagnosis and treatment process. Second, the specific characteristics of the participants' individual culture and their view of pain and cancer could result in different cancer pain experiences even among white cancer patients. Third, the participants complained that women's pain was not taken seriously by healthcare providers. Fourth, the participants reported highly individualized pain experience with emotional pain. Finally, the participants wanted to have a control of their own pain management process. Based on the findings, implications for nursing research and practice are proposed. PMID- 17135818 TI - Cervical cancer screening--"For better or worse...": women's experience of screening. AB - In Sweden, the population-based cervical cancer screening program has been in progress since the 1970s and is directed toward women between 23 and 60 years of age. The aim of this study was to explore women's experience of cervical screening after being diagnosed with cancer. A qualitative study inspired by Grounded Theory was used. Eleven women were interviewed. The interviews were analyzed using the constant comparative method. "Screening-For Better or Worse..." was identified as the core category, around which the categories "Unawareness," "Trust," "Search for Understanding," and "Making the Invisible Visible" were integrated and on which the conceptual model was built. The experience of screening had both a positive and a negative dimension for the women stricken by cervical cancer. The women had a positive experience of screening as such and they believed in its benefits. However, many women felt deceived and, because of their unawareness, questions arose for which they searched for understanding. The women requested adequate and understandable information. Their trust remained because the contact with healthcare professionals involved in screening and in the follow-up program had been reassuring. PMID- 17135819 TI - Rural women's perception of breast cancer and its early-detection measures in Ibadan, Nigeria. AB - Breast cancer is the leading cause of cancer death in women in many parts of the world and is the most common cancer in women in Nigeria. However, it lends itself to early detection and, subsequently, early treatment, if women use early detection measures. It is pertinent to study rural women's perception of breast cancer and its early-detection measures because their perception would influence their use of early-detection measures of breast cancer. This is a descriptive study that assessed rural women's perception of breast cancer. Data from 2 rural health districts in Ibadan, Oyo state of Nigeria, were collected through a structured questionnaire. Four hundred seven women were randomly selected for the study. Results showed that 66.2% of the respondents considered that breast cancer is more severe than other forms of cancer. Respondents' perception of risk of developing breast cancer was low, as 64.8% rated themselves 1, on a scale of 1 to 9 (where 1 = does not perceive herself to have cancer; 9 = very much perceives herself to have cancer). Respondents' perceived cause of breast cancer included "putting money in brassiere" and attack from the enemy, among others. None of the respondents identified early detection as an advantage of breast self examination. Swelling was the most acknowledged early-warning sign. Findings of this study indicate that there is a need for an interventional study to enhance the awareness of breast cancer and its early-detection measures among the rural population to influence early detection of breast cancer and subsequently reduce morbidity and mortality among them. PMID- 17135820 TI - Course of fatigue between two cycles of adjuvant chemotherapy in breast cancer patients. AB - The purpose of this study was to determine the course of fatigue in patients with breast cancer between 2 cycles of adjuvant chemotherapy, from the day of administration until the day of the next infusion. In a prospective cohort study, a sample of 151 patients with breast cancer receiving adjuvant chemotherapy was recruited from 6 hospitals in mainly the south of the Netherlands. Patients reported their experience of fatigue in a diary, the Shortened Fatigue Questionnaire, on a daily basis between the third and fourth treatment with adjuvant chemotherapy. Patients were treated with either a doxorubicin containing schedule or with cyclophosphamide, methotrexate, and 5-fluorouracil (CMF, 28 days). In the 28-day regimens, infusions were given on day 1 and day 8. The days after completion of the third and the start of the fourth treatment with chemotherapy were statistically analyzed. We tested the hypothesis that the maximum fatigue score occurs in the first 4 days after treatment. The mean age of the sample was 47.2 years (SD = 8.8). Most women (84%) were married or lived together with a partner. The majority (80%) of all patients had been diagnosed with stage II breast cancer. The division between mastectomies (47%) and lumpectomies (52%) was approximately equal. Sixty percent of the patients received radiotherapy before the third treatment with chemotherapy and/or in the period they kept the diary. A chaotic pattern of fatigue between the 2 cycles of chemotherapy emerged. Smooth (splines) curves showed an average highest level of fatigue on day 3 from the start. For the 28-day regimens, another distinct peak was seen around day 11. A relatively larger number of patients experienced peak fatigue levels before day 5. The course of fatigue in the CMF group was significantly different compared with the doxorubicin regimens. The fatigue peak in the CMF group was lower. Women taking cyclophosphamide orally experienced the peak level of fatigue significantly later. Influences of other variables were not observed in any chemotherapy group. Cancer-related fatigue has a chaotic nature. The first days after treatment with chemotherapy are the worst. The type of chemotherapy has a significant impact on the course of fatigue. Improved understanding of the nature and course of fatigue could equip healthcare providers better for informing patients about what they may expect. Future research should include interventions aimed at reducing or coping with fatigue. PMID- 17135822 TI - Posttreatment breast cancer surveillance and follow-up care experiences of breast cancer survivors of African descent: an exploratory qualitative study. AB - Breast cancer survivors are at considerable risk for breast cancer recurrence and at higher risk of developing a new breast cancer compared with women never diagnosed. It is recommended that survivors undergo careful breast cancer surveillance as cancers detected early are more treatable. However, data indicate that surveillance among African American survivors, particularly mammography, is lower than that of white survivors. There is little published work focusing on general experiences of posttreatment breast cancer surveillance among survivors of African descent. In the current qualitative pilot study, key informant interviews were conducted in order to explore the following: (1) the extent of posttreatment surveillance information provided to or obtained by survivors of African descent; (2) the actual follow-up care received by survivors in the past year; and (3) factors that are either motivators of or barriers to care. Participants were 10 African American and African Caribbean breast cancer survivors. Survivors reported a number of factors that motivated them in obtaining follow-up care: a desire to maintain good health, concern about recurrence, support from healthcare providers, familial relationships, relationships with other survivors, and religious/spiritual faith. Survivors also reported barriers to care: fear of recurrence, low support from family and friends, lack of information about posttreatment follow-up care, and medical care costs. These results represent formative work that may inform similar studies examining factors in breast cancer surveillance and follow-up care in larger samples of survivors of African descent. PMID- 17135823 TI - Effects of mutual dyad support on quality of life in women with breast cancer. AB - This study examined the effects of a mutual dyadic support intervention between survivors and women newly diagnosed with breast cancer on the quality of life and interpersonal relationships of the participants. This matched-pairs, baseline postintervention study included 31 survivors and 31 newly diagnosed women with stage I or II breast cancer. The women met in mutually supportive, self-directed dyads at least twice a week for 8 weeks. Instruments included contact Logs and Meaningful Observations Journals. Content analysis of the Logs showed that most dyad interactions were by telephone. Most participant dyadic conversations focused on the health and functioning of newly diagnosed partners, with survivors providing informational and emotional support. Meaningful Observations Journal thematic analysis supported the social exchange theory to describe dyad partner relationships. Most dyadic relationships were supportive, some were reciprocal, and some experienced conflict. Themes varied little between groups and from baseline to postintervention. Themes across time were support of others, health, changing priorities, and faith. Further quantitative study is needed to examine the effects of a dyad support intervention on quality of life in more diverse populations of women with breast cancer. Nurses need to evaluate the existing social-network support systems of patients with breast cancer and consider the need for dyadic peer support. PMID- 17135824 TI - Evaluation of the complexity of postoperative care following breast and gynecologic cancer surgery. AB - The purpose of this study was to evaluate the complexity of postoperative care required by women who had undergone breast or gynecologic cancer surgery, using the Therapeutic Intervention Scoring System-28. An observational, longitudinal study was carried out on 83 women admitted postoperatively to the Intensive Care Unit of the Center for Women's Integrated Healthcare. The results of this study show that women diagnosed with gynecologic cancer had a significantly higher mean Therapeutic Intervention Scoring System-28 score compared with women who had been diagnosed with breast cancer (P = .01). Women who underwent gynecologic surgery had a significantly higher mean Therapeutic Intervention Scoring System-28 score than women who had breast surgery (P = .03). Most of the activities performed by staff during the postoperative intensive care unit period involved only basic care procedures. Only advanced age and hypertension were related to death during the postoperative period. In conclusion, from the nursing standpoint, the complexity of care required in the intensive care unit during the postoperative period was moderate. Women aged 80 years or older were at the highest risk of death during this period. PMID- 17135837 TI - Infection control for the family of the home hospice patient. PMID- 17135838 TI - Integrating standards into practice. Revised standards for home care infusion: what has changed? AB - In 2006, the latest revision of the Infusion Nursing Standards of Practice was published by the Infusion Nurses Society. The Standards define the scope of practice and educational requirements for nurses who practice in infusion therapy in all healthcare settings. This article provides a description of the Standards, including a discussion of 12 Standards to illustrate their application in home care. PMID- 17135839 TI - Research on falls prevention and physical activity in older adults and a notice of a new web-based quality system by the agency for healthcare research and quality. PMID- 17135840 TI - Responding to patient-initiated humor: guidelines for practice. AB - Over the past few decades, increasing evidence has shown the beneficial effects of humor. The use of caregiver-initiated humor as an intervention in healthcare settings has both physiological and emotional benefits. Little has been written, however, about another very important aspect of humor, patient-initiated humor. When patients use humor to relieve their feelings of stress, uncertainty, or embarrassment, they are trying to communicate with their caregiver. This use of humor by patients is not to "make light" of the situation, but rather a way to reduce their feelings of dehumanization. Humor is an interactive process of sharing and an important aspect of communication. Patients will observe the caregiver for a response. An open, accepting response signals understanding; a negative or null response, however, may serve to isolate the patient. The guidelines discussed in this article for recognizing, interpreting, and responding to patient-initiated humor will help home care and hospice nurses to foster increased open patient/caregiver communication and create a supportive humanistic atmosphere for patient care. PMID- 17135842 TI - The benefits of diabetes self-management education of the elderly veteran in the home care setting. PMID- 17135844 TI - A pathway for moral reasoning in home healthcare. AB - Ethical issues arise in the practice of infusing nursing, and when they do, the home care nurse can find guidance in the code of ethics. This article addresses the importance of the code of ethics and introduces a framework for decision making to assist the home care nurse in determining a course of action in the face of ethical quandaries. PMID- 17135845 TI - Changing culture in the home health setting: strategies for success. AB - Organizational culture is generally defined as the internal attributes of the staff, such as their values, beliefs, and attitudes. Although technically accurate as a definition, personal attributes defy direct intervention, leading some to question whether it is possible to change culture. It is proposed that it is possible to change the personal internal attributes that define organizational culture by changing the characteristic structures and behaviors of the organization that shape those attributes. This model, called the Quality Capability Model, creates an approach to culture change that accommodates the unique features of home health. PMID- 17135847 TI - Professional boundaries in home care. PMID- 17135848 TI - Caring for patients of diverse religious traditions: Evangelical "Born Again" Christians. PMID- 17135849 TI - Home infusion therapy. First things first: the patient and the prevention of central catheter infections. AB - During the past seventeen years, I've had the privilege of working as a home healthcare nurse in two rural counties in the Upper Peninsula of Michigan. In the early 1990s, it was rare that our agency received referrals for IV antibiotics, opiate infusions for pain management or for total parenteral infusion (TPN) to be administered at home. Most patients stayed in the hospital for infusion therapy. Today, as more healthcare treatments are being shifted from hospital to outpatient or home care settings, referrals for home infusion have become more common in our area as well as across the nation (Jarvis, 2001). I'll never forget my first patient whom I cared for with home infusion therapy for pain management. I learned a great deal from Sally, and to this day I always remember that I can make a real difference in patient outcomes when I keep--"First Things First": the patient and infection prevention--when caring for any of my patients. PMID- 17135850 TI - National policies need "nursing intervention". PMID- 17135851 TI - My say: a caregiver's view of the world. The pain, the joy, and the blessing of caregiving, in-home. PMID- 17135853 TI - The American Heart Association CPR Anytime Program: the potential impact of highly accessible training in cardiopulmonary resuscitation. PMID- 17135854 TI - A systematic approach to improve lipids in coronary artery disease patients participating in a cardiac rehabilitation program. AB - PURPOSE: To determine the effectiveness of an intervention, directed toward the primary care physician (PCP), to improve the number of patients treated to low density lipoprotein cholesterol (LDL-C) goal in a cardiac rehabilitation (CR) population. METHODS: A pre-post intervention cohort comparison using data collected from participants in a CR program with LDL-C > or =100 mg/dL at entry. The control cohort participated in CR between 1/00 and 10/02, 41.5% (n = 178) had an entry LDL-C > or =100 mg/dL. The intervention cohort participated in CR between 10/03 and 1/05, 26.4% (n = 67) had an entry LDL-C > or =100 mg/dL. The intervention group had identical treatment as the control group as well as the following: each participant with an LDL-C > or =100 mg/dL in the intervention cohort had an entry letter sent to his or her cardiologist and PCP from the programs Cardiology Medical Director, detailing the lipid goals and therapeutic options. In addition, monthly faxes on progress toward lipid goals were sent to the PCP. RESULTS: The control cohort was less likely to achieve LDL-C goal compared with the intervention cohort (43% vs 67%, respectively; P = .001). A patient was also less likely to have a lipid medication change during CR in the control group compared with the intervention group (29% vs 42%, respectively; P = .05). CONCLUSION: Use of systematic reminders directed at the PCP during CR can substantially increase the percentage of patients achieving nationally recognized LDL-C goals. PMID- 17135856 TI - Closing the cholesterol treatment gap: cardiac rehabilitation can make a difference. PMID- 17135857 TI - Five-year changes in North Carolina outpatient cardiac rehabilitation. AB - PURPOSE: The purpose of this study was to describe cardiac rehabilitation programming, barriers to participation, and reasons for dropout in North Carolina from a program director's perspective and to compare those results with those of a similar statewide survey conducted 5 years earlier. METHODS: In 1999 and 2004, a survey was mailed to all North Carolina program directors of outpatient cardiac rehabilitation programs. The response rate was 85% (61/72) in 1999 and 79% (61/77) in 2004. RESULTS: More than 85% of North Carolinians older than 40 years lived within a 15-mile buffer of an outpatient cardiac rehabilitation program in 2004. Most programs were staffed with personnel trained in nursing, exercise physiology, and nutrition in 1999 and 2004. Women and African Americans remained disproportionately underrepresented as participants in the program for both years. In 2004, approximately one third of cardiac rehabilitation programs reported having a referral to rehabilitation on the hospital discharge plan for myocardial infarction and coronary artery bypass surgery. In 1999 and 2004, the most frequently reported barrier to participation remained financial, followed by lack of interest or motivation and workplace conflicts. Work conflicts, lack of interest, and comorbidities were the most frequently reported reasons for dropping out from cardiac rehabilitation programs in both 1999 and 2004. CONCLUSIONS: Increasing participation in cardiac rehabilitation programs by addressing barriers at multiple levels may facilitate greater patient participation. This statewide survey could be used in other states as a surveillance tool, to track changes in rehabilitation over time from a program director's perspective. PMID- 17135858 TI - Correlates of physical activity change in patients not attending cardiac rehabilitation. AB - OBJECTIVE: Limited research has identified theoretical correlates of physical activity (PA) change in patients not receiving cardiac rehabilitation. The purpose of the present study was to determine whether changes in self-efficacy, PA intention, perceived severity and susceptibility, and PA benefits/barriers were associated with changes in PA over a 12-month period in these patients. METHODS: Patients (N = 555) not attending cardiac rehabilitation completed a psychosocial questionnaire in hospital and 6 and 12 months after hospitalization for a cardiac event. RESULTS: Hierarchical regression analyses showed that the increase in PA from baseline to 6 months was significantly related to an increase in self-efficacy and PA intentions and a decrease in the impact of health-related barriers. Furthermore, the decrease in PA from 6 to 12 months was significantly related to a decrease in health-related benefits and PA intentions and an increase in time and health-related barriers. Finally, the increase in PA from baseline to 12 months was significantly related to an increase in health-related benefits and intentions and a decrease in health-related barriers. CONCLUSIONS: Changes in PA levels over a 12-month period were associated with changes in various theoretical variables. Interestingly, the associations among these variables with PA varied as a function of time after hospitalization. PMID- 17135859 TI - Standardizing and predicting results from cardiopulmonary exercise testing in patients with heart failure. AB - PURPOSE: Cardiopulmonary exercise testing is a common prognostic tool in heart failure, yet it is not standardized. The purpose of this study was to evaluate a means of standardizing oxygen consumption (VO(2)) measurement and to evaluate the ability to predict peak VO(2) from submaximal exercise. METHODS: Fifty consecutive exercise tests with a respiratory exchange ratio > or =1.10 were evaluated. VO(2) was graphed against respiratory exchange ratio and the peak VO(2) was determined with logarithmic, linear, power, and exponential regression lines. To predict a peak VO(2), each patient's submaximal exercise data (respiratory exchange ratio < or =0.98) were fitted to each regression line. The mean of the last 30 seconds of un-averaged breath-by-breath data was used as the reference value. Peak VO(2) assessments are also provided from the metabolic cart, a rolling time average, and the graphical method. RESULTS: Logarithmic regression best standardized peak VO(2). Mean absolute bias (mL x kg x min) was 0.60 +/- 0.44 for logarithmic, 0.61 +/- 0.47 for linear, 0.85 +/- 0.67 for power, and 1.44 +/- 2.22 for exponential. The mean absolute bias between the peak logarithmic predicted VO(2) and the reference peak VO(2) was 1.62 +/- 1.20 mL x kg x min (9.5% of the peak VO(2)). CONCLUSION: Among the methods studied, logarithmic regression analysis was the best method to standardize and predict peak VO(2) in this cohort of patients with heart failure. PMID- 17135860 TI - The effect of handrail support on oxygen uptake during steady-state treadmill exercise. AB - RATIONALE: Heart rate (HR) and oxygen consumption (VO(2)) are indicators of the intensity of exercise. Handrail support has been shown, during maximal treadmill testing, to blunt HR and VO(2) responses at a particular speed and grade, resulting in an increased treadmill time and overprediction in aerobic capacity. OBJECTIVES: This study was designed to determine if handrail support would similarly blunt HR and VO(2) responses during steady-state treadmill exercise at intensities typical of exercise training. METHODS: Healthy volunteers (age, 38-60 years; N = 10) performed maximal treadmill exercise to define VO2max (35.4 +/- 6.5 mL kg(-1) min(-1)) and ventilatory threshold (26.4 +/- 5.8 mL kg(-1) min( 1)). They also performed 3 random steady-state exercise bouts including free arm swing, handrail support-resting, and handrail support-gripping (HRS-G). Each test consisted of three 5-minute stages with intensity levels corresponding to 75%, 85%, and 95% of the speed and grade at ventilatory threshold. RESULTS: There were significant (P < .05) differences in HR and VO2 at the 75%, 85%, and 95% ventilatory thresholds in HRS-G (108, 114, and 121 beats min and 17.2, 18.0, and 20.6 mL kg min, respectively) versus handrail support-resting (114, 126, and 137 beats min and 19.5, 21.8, and 23.9 mL kg min, respectively) and HRS-G versus free arm swing (120, 130, and 142 beats min and 20.3, 22.8, and 26.1 mL kg min, respectively). Rating of perceived exertion was significantly (P < .05) different between HRS-G (1.8, 2.4, and 3.1) and free arm swing (2.2, 2.9, and 3.6) at all intensities and between HRS-G (2.4 and 3.1, respectively) and handrail support resting (3 and 3.7, respectively) at the 85% and 95% ventilatory thresholds. CONCLUSION: Gripping and, to a lesser degree, resting the hands on the handrails during steady-state treadmill walking will blunt responses during exercise training and may result in less predictable exercise program. PMID- 17135861 TI - A new functional status outcome measure of dyspnea and anxiety for adults with lung disease: the dyspnea management questionnaire. AB - PURPOSE: To develop and psychometrically test the Dyspnea Management Questionnaire (DMQ), a new multidimensional measure of dyspnea in adults with chronic obstructive lung disease. PARTICIPANTS: Eighty-five participants were recruited with diagnoses of chronic obstructive pulmonary disease (COPD, n = 73) and asthma (n = 12). The total sample was predominately female (65%) and married (34%), with 64.9% white and a mean age of 75 years (SD = 9.6, n = 76), diagnosed with pulmonary disease 4.8 years ago (SD = 4.4), 32% requiring the use of supplemental oxygen. Participants were also African American (29.9%), Asian (2.6%), and Hispanic (2.6%); n = 77. METHODS: An initial item pool of 74 items was drawn for the DMQ aided by qualitative interview data, literature review, and pilot testing with 3 adults with COPD. Several analyses were used to reduce the item pool. An interdisciplinary panel of 12 experts evaluated the content validity of the DMQ items. To evaluate test-retest reliability, respondents with stable COPD (n = 26) completed the questionnaire twice within a mean interval of 18 days (SD = 7.17). The DMQ was compared with the Medical Outcomes Study 12-Item Short-Form (SF-12) Health Survey, the Seattle Obstructive Lung Disease Questionnaire, and the Hospital Anxiety and Depression Scale. RESULTS: The resulting DMQ is a 30-item scale that measures 5 conceptually derived dimensions: dyspnea intensity, dyspnea-related anxiety, fearful activity avoidance, self efficacy for activity, and satisfaction with strategy use. It has a 7-point Likert-type scale and third Flesch-Kincaid reading grade level. A panel of 12 experts supported the content validity of the DMQ. It showed high internal consistency (alpha = .87 to .96) and test-retest reliability over 2.5 weeks (intraclass correlation coefficient = 0.71 to 0.95). Dyspnea intensity, dyspnea related anxiety, and fearful activity avoidance subscales of DMQ-30 and composite score were moderately to highly correlated with 3 Seattle Obstructive Lung Disease Questionnaire dimensions (r = 0.44-0.83), Medical Outcomes Study 12-Item Short-Form scales (r = 0.41-0.57), and Hospital Anxiety and Depression Scale Anxiety (r = -0.59 to -0.65). Two of DMQ's subscales, self-efficacy for activity and satisfaction with strategy use, correlated mildly with Seattle Obstructive Lung Disease Questionnaire (r = 0.28 and 0.27, respectively). Some very low correlations for DMQ-30's satisfaction with strategy use compared with the Medical Outcomes Study 12-Item Short-Form provided preliminary support for its divergent construct validity. The DMQ-30 discriminated adults with COPD requiring supplemental oxygen from those not requiring it. CONCLUSIONS: The DMQ addresses the need for a more comprehensive, multidimensional assessment of dyspnea, especially for anxious patients with COPD, in order to better guide the appropriate application of dyspnea management interventions and measure pulmonary rehabilitation outcomes. The DMQ can help add insights into the benefit of adjunctive therapies such as psychoeducation, controlled breathing strategies, and cognitive-behavioral approaches in pulmonary rehabilitation for anxious patients with COPD. PMID- 17135862 TI - Does sitting posture in chronic obstructive pulmonary disease really matter? An analysis of 2 sitting postures and their effect [corrected] on pulmonary function. AB - PURPOSE: The purpose of this study was to investigate changes that occur in pulmonary function when postural changes in the sagittal plane are made in a seated position in patients diagnosed with chronic obstructive pulmonary disease (COPD). METHODS: Fourteen patients diagnosed with COPD participated in this study. Standard spirometric measurements (minute ventilation, forced vital capacity, and forced expiratory volume in 1 second) were taken for each patient in each of 2 sitting postures: slumped and upright. Breathing frequency, heart rate, and blood oxygen saturation were also recorded for each of the 2 postures. Patients assumed each posture for 5 minutes before any measurements were taken, after which measurements were recorded each minute for an additional 5 minutes. RESULTS: A 2-factor (posture and time) analysis of variance with repeated measures on both factors was used to analyze the data. There were no significant differences between the means for heart rate, blood oxygen saturation, and breathing frequency. Paired t tests likewise did not reveal any significant differences between the slumped and upright positions for forced expiratory volume in 1 second, forced vital capacity, and minute ventilation. CONCLUSIONS: These results suggest that there are no differences in measures of pulmonary function (minute ventilation, forced vital capacity, and forced expiratory volume in 1 second) and breathing frequency, heart rate, and blood oxygen saturation between slumped and upright sitting in patients with COPD. Based on this evidence alone, it may be inappropriate to instruct a patient with COPD to sit upright to improve respiratory function. However, further study is warranted before any definite recommendations can be made regarding sitting posture and respiratory performance in individuals with COPD. PMID- 17135870 TI - Silence kills ... Dialogue heals. PMID- 17135871 TI - Shining the light on the "800-lb Gorilla" of professional rivalry in case management. PMID- 17135872 TI - Quantifying case management workloads: Development of the PACE tool. AB - In 1999, the Clinical Resource Management (CRM) group at Saint Alphonsus Regional Medical Center in Boise, Idaho, implemented a specialty-based nursing case management model for individual caseload assignment of all inpatient admissions. This model assigns caseloads by proportion determined by physician specialty type. This model does not consider patient acuity and may have weaknesses associated with self-report. In addition, time requirements for effective case management outcomes are not always reflected in admission volumes alone. This article describes the development and pilot testing of the Patient Acuity Case management Evaluation (PACE) measurement tool. Five phases of methodological development are discussed: (1) conceptualization phase, (2) content development, (3) Delphi technique using expert opinion and critique, (4) inter (intrarater reliability testing, and (5) pilot testing of acuity tool. Examples of instrument development throughout these phases are included. This instrument is expected to provide verifiable measurement of individual and group case management workload. PMID- 17135874 TI - Using continuous quality improvement to implement evidence-based medicine. AB - The importance of implementing evidence-based medicine is being driven by public reporting of outcome data and linking these measures to reimbursement. Most hospitals are faced with many challenges in gaining sponsorship, staffing, creating tools, and reporting of evidence-based outcome measures. This article describes the use of the SSM Health Care (SSMHC) Continuous Quality Improvement model in implementing evidence-based practices at SSM DePaul Health Center, a community hospital member of SSMHC, including successes, opportunities for improvement, and lessons learned. Specifically, the article includes two different processes for data collection and interventions with staff, process requirements for each, and outcome data associated with each model. PMID- 17135876 TI - The Caring Connections Project: Providing palliative care to Medicaid patients with advanced cancer. AB - Palliative care, with its focus on symptom management, patient-centered goals, preparation for life's end, and preservation of quality of life in the face of advancing illness, is a rapidly advancing component of mainstream American medicine. Yet, access to palliative care is often lacking in the community setting and may be further hindered by the presence of healthcare disparities that impact the poor. This article presents a unique approach to assuring the availability of palliative care to Medicaid patients receiving case management services. This descriptive article describes the evolution of a palliative care management pilot program, the Caring Connections Program, beginning with the initial planning and progressing through implementation and provision of services to 56 persons. "Lessons learned" are shared to enable other providers to develop similar programs with success. Patient profiles and intervention strategies are offered to illustrate the work accomplished. PMID- 17135877 TI - Brief intervention for weight management--PART II. PMID- 17135878 TI - What's next in disease management? PMID- 17135880 TI - Patient safety in healthcare: What case managers can do. PMID- 17135893 TI - Managing postpolio syndrome pain. PMID- 17135894 TI - Assisting with thoracentesis. PMID- 17135895 TI - Time to prevent injuries from postimmunization syncope. PMID- 17135896 TI - Perineal dermatitis or pressure ulcer: how can you tell? PMID- 17135897 TI - Are NSAIDs safe for arthritis pain? PMID- 17135899 TI - Is your patient susceptible to malignant hyperthermia? PMID- 17135904 TI - Getting the most from an admission interview. PMID- 17135917 TI - Understanding the 12-lead ECG, part II. Learn to recognize bundle-branch blocks, myocardial infarction, and common dysrhythmias. PMID- 17135919 TI - In her footsteps... AB - It was a special day for my patient, who was nearly 100. Today I pushed her in a wheelchair, but years ago, she'd helped pave my way. PMID- 17135920 TI - Hypothermia: a hazard for all seasons. AB - Find out how to respond when your patient's body temperature takes a dive, no matter what the cause. PMID- 17135921 TI - Walk a fine line if your patient wants to leave AMA. AB - What should you do? Take this attorney's advice to protect yourself and your facility from legal repercussions. PMID- 17135922 TI - Responding to foreign-body airway obstruction. AB - Learn how the guidelines for quickly helping a choking adult have changed. PMID- 17135923 TI - Supporting nutrition with T.E.N. or T.P.N. AB - Learn the pros and cons of each type of specialized nutrition support and how to prevent problems related to therapy. PMID- 17135924 TI - Postoperative wound care. PMID- 17135925 TI - New weapons to snuff out kidney cancer. AB - Learn how to assess and care for a patient with this often-hidden cancer. Then get the scoop on two newly approved drugs to treat advanced stages of the disease. PMID- 17135935 TI - . . . About factor V Leiden. PMID- 17135936 TI - ECG equipment: wired for infection? PMID- 17135937 TI - Groin hematoma following percutaneous coronary intervention. PMID- 17135939 TI - Anatomy and biomechanics of the lateral side of the knee. AB - The posterolateral corner (PLC) of the knee is a critical element for a functional lower extremity. It consists of an array of complex ligamentous and musculotendinous structures. The primary function of the PLC is to resist varus and external rotation and posterior translation of the tibia. Injuries to these structures can cause significant disability and compromise activities of daily living and work, recreational, and sporting activities. A thorough understanding of the complex anatomy and biomechanics of the PLC will aid the clinician in this challenging diagnostic and therapeutic problem. The first section of this paper describes the anatomy of the PLC of the knee focusing on the intricate insertion sites of the individual structures. The second section discusses how the anatomy influences the biomechanics of the PLC. PMID- 17135940 TI - Physical examination and imaging of the lateral collateral ligament and posterolateral corner of the knee. AB - Diagnosis of posterolateral corner injuries can be challenging. Understanding the mechanism of injury in combination with careful history taking and a thorough physical examination are crucial. Clinical findings may be subtle, especially in the acute setting. Specialized tests such as external rotation recurvatum test, posterolateral drawer test, reverse pivot shift test, and dial test are particularly helpful. A characteristic radiographic finding is the arcuate sign, whereas medial Segond fractures can be associated with posterolateral corner injuries. Magnetic resonance imaging is most useful when performed with a high strength magnet utilizing a coronal oblique technique. Prompt and accurate diagnosis of posterolateral corner injuries allows the orthopedist to initiate the appropriate repair and reconstruction procedures to assure optimum treatment results. PMID- 17135941 TI - Nonsurgical management of lateral side injuries of the knee. AB - Non-surgical management of posterolateral corner (PLC) knee injuries is reserved for specific isolated mild to moderate injuries. There has been a relative scarcity of studies discussing non-surgical management existing secondary to the relative rarity of isolated PCL injuries. In these specific cases, a few studies have shown non-surgical management to result in satisfactory outcomes. This review of the literature outlines the outcomes and treatment options for posterolateral corner (PLC) knee injuries, which is based on the grade of the injury. However, no matter what the grade of injury, it is crucial to rule out other associated deficiencies before undertaking a nonoperative approach in the management of the posterolateral corner of the knee. PMID- 17135942 TI - Surgical treatment of acute lateral collateral ligament and posterolateral corner injuries. AB - Early surgical treatment of injuries to the posterolateral corner of the knee is recommended to promote rehabilitation for normal function of the knee. To optimize the chance at primary repair, surgery should be performed within 2 weeks; otherwise, reconstruction is the most viable option. During operative treatment, each component of the posterolateral corner is assessed individually, with primary repair performed if amenable to repair. Additional techniques, such as augmentation, advancement, or reconstruction may be needed if the primary repair is tenuous or the tissues damaged. PMID- 17135943 TI - Novel approach for reconstruction of the posterolateral corner using a free tendon graft technique. AB - Injuries to the lateral collateral ligament (LCL) and posterolateral corner of the knee, particularly when combined with anterior cruciate or posterior cruciate ligament injuries, can result in profound symptomatic knee instability. Although many surgical improvements have been made in the reconstruction of anterior and posterior cruciate ligament injuries, reconstruction of the posterolateral corner has had less predictable results, with residual pathologic laxity especially in the chronic situation. This has stimulated many surgeons to recommend acute repair of posterolateral knee injuries. This article will briefly review the relevant surgical anatomy, present a summary of current reconstructive techniques for the posterolateral corner, and describe our preferred method for anatomic reconstruction of the posterolateral corner for chronic instability of the knee by recreating the LCL and popliteofibular ligament using either autogenous or allograft soft tissue and an interference screw technique. We do not use a transtibial tunnel but re-orientate the transfibular tunnel and utilize 2 femoral tunnels an the attempt to recreate the LCL and popliteus tendon. In a small clinical series, this has proven to restore varus rotation and external rotation patholaxities with a high degree of predictability. PMID- 17135944 TI - Surgical treatment of lateral posterolateral instability of the knee using biceps tendon procedures. AB - Posterolateral instability (PLI) is common with posterior cruciate ligament tears and is less common with anterior cruciate ligament tears, and isolated PLI is rare. There are varying degrees of PLI with respect to pathologic external tibial rotation, and varus laxity. Surgical treatment of PLI must address all components of the PLI (popliteus tendon, popliteofibular ligament, lateral collateral ligament, and the lateral-posterolateral capsule), and other structural injuries. Successful posterior cruciate ligament and anterior cruciate ligament surgery depends upon recognition and treatment of posterolateral corner injuries. PMID- 17135945 TI - Surgical treatment of chronic posterolateral rotatory instability of the knee using capsular procedures. AB - Chronic posterolateral rotatory instability of the knee was introduced as a diagnostic classification by Hughston and associates in the early 1970s and occurs as a result of dysfunctional healing of a strained arcuate complex, causing a patulous posterolateral capsuloligamentous complex to exist. This capsular redundancy allows varying degrees of recurvatum and adduction instability during single-limb stance. Eradication of the redundant pouch with a capsular shift-type reconstruction has been shown to eliminate the instability and hold up under long-term follow-up studies. Understanding the complex normal and injured anatomy of the posterolateral corner of the knee is essential for effective reconstruction of this instability. PMID- 17135946 TI - Complications in the treatment of medial and lateral sided injuries of the knee joint. AB - Complications may result from nonoperative and surgical management of medial or lateral sided knee injuries. Because these injuries are often associated with injury to the medial or lateral meniscus and the anterior or posterior cruciate ligaments (ACL and PCL, respectively), injury to these structures will also be considered. We group these complications in 3 categories: (1) complications associated with either operative or nonoperative management, (2) intraoperative complications, and (3) postoperative complications. Greater understanding of complications of medial and lateral knee ligament injuries may result in improved clinical outcomes. PMID- 17135948 TI - Evaluation and treatment of medial collateral ligament and medial-sided injuries of the knee. AB - Injuries to the medial side of the knee are not always isolated injuries of the superficial medial collateral ligament. Medial-sided injuries can also involve the deep medial collateral ligament, the posteromedial corner, or the medial meniscus. Magnetic resonance imaging is a useful adjunct to the physical examination; however, the extent of medial-sided injuries is frequently underappreciated on these images. An understanding of the anatomy and biomechanics of the medial side of the knee and a thorough physical examination aids the physician in determining the full extent of injury and helping the physician to treat each unique injury pattern. PMID- 17135949 TI - Physical examination and imaging of the medial collateral ligament and posteromedial corner of the knee. AB - The medial side is one of the most commonly injured areas of the knee. The anatomy, diagnosis, and treatment of medial collateral ligament and posteromedial corner (PMC) injuries can be challenging. Understanding the mechanism of injury and performing a thorough physical examination and radiographic evaluation is essential. Frequently, there are concomitant meniscal and other ligament injuries associated with medial-side injuries. Correct identification of all injured structures is important, as missed diagnoses can lead to significant disability. Unrecognized PMC injuries have been implicated in anteromedial rotary instability and failed anterior cruciate ligament reconstructions. Valgus stress testing is the cornerstone for the identification of medial-side knee injuries. Coronal sequences from magnetic resonance imaging are the most useful tool to view the medial collateral ligament and posteromedial structures. Stress x-rays and ultrasound may also be helpful. Thorough physical examination and imaging of injuries to the PMC should dictate the appropriate treatment for optimal results. PMID- 17135950 TI - The non-surgical management of isolated medial collateral ligament injuries of the knee. AB - The medial collateral ligament is the most commonly injured ligament in the knee. All isolated grade I and II tears and most grade III tears can be treated nonoperatively with a supervised, functional, rehabilitation program. Good to excellent results can be expected with return to full preinjury activity level being the norm. PMID- 17135951 TI - Surgical treatment of acute medial collateral ligament and posteromedial corner injuries of the knee. AB - The medial collateral ligament of the knee is unique in that it has shown the capacity to heal with conservative measures. As a result, nonoperative treatment is often successful in returning patients back to previous levels of activity and function. However, specific surgical indications do exist for certain isolated and many combined medial collateral ligament injuries. Strict adherence to both nonoperative and operative principles allows for optimum treatment in most instances. PMID- 17135952 TI - Evaluation and treatment of chronic medial collateral ligament injuries of the knee. AB - Injuries to the medial collateral ligament (MCL) can occur as isolated injuries or in conjunction with injuries to other structures about the knee. Most grade I and II MCL injuries without meniscal avulsion, alone or in combination with anterior or posterior cruciate ligament injuries, can be treated nonoperatively. Grade III or complete tears also can be treated nonoperatively, but only after careful exclusion of any associated injuries that may require surgical treatment. Treatment recommendations also have been based on the location of the MCL tear and the associated injuries. Surgical treatment may include reconstruction of the anterior and posterior cruciate ligaments with primary repair of the MCL. Chronic medial knee injuries often are associated with concomitant ligament injuries, which also must be treated. Treatment options include nonoperative (bracing, activity modification, and rehabilitation) and operative reconstruction. PMID- 17135953 TI - Surgical treatment of chronic posteromedial instability using capsular procedures. AB - Patients with posteromedial corner injuries of the knee present a significant problem to the clinician. Often the symptoms of anteromedial rotatory instability may be overlooked due to concurrent anterior cruciate ligament and/or posterior cruciate ligament injuries. When addressing concurrent ligamentous injuries to the knee, the clinician may fail to isolate anteromedial rotatory instability on physical examination and imaging studies may not specifically identify damage to posteromedial corner structures. This unrecognized and untreated damage results in residual functional laxity and can potentially lead to chronic and progressive instability of the knee. This paper contains descriptions of 2 surgical techniques that address posteromedial instability. Both techniques require an understanding of posteromedial corner knee anatomy, careful preoperative planning, and extensive postoperative rehabilitation. PMID- 17135954 TI - Role of high tibial and distal femoral osteotomies in the treatment of lateral posterolateral and medial instabilities of the knee. AB - Mechanical alignment has been overlooked as an important contributor to knee stability with respect to collateral ligament laxity. The detrimental effects of varus or valgus limb malalignment become more obvious when the restraining force is lost because of ligamentous injury especially on the medial or the lateral/posterolateral side. Even with repair or reconstruction of these injuries, with repetitive load, the ligament will eventually fail to restore its structure and strength, resulting in failure and secondary restraint laxity. We have found that realignment of the limb is the most important factor in restoring a functional limb. After realignment, joint laxity, if persistent, can be addressed successfully with ligamentous reconstruction. Diagnosis and treatment of limb malalignment cannot be ignored in the management of chronic ligamentous instabilities, especially those with prior failed reconstruction. Our approach to these difficult problems and the preferred techniques of osteotomies on both tibial and femoral sides are described. PMID- 17135955 TI - Conservative and postoperative rehabilitation of isolated and combined injuries of the medial collateral ligament. AB - Injuries to the medial collateral ligament (MCL) are very common and there seems to be a consensus supporting the conservative management of grade I and II tears. Grade III tears are also usually treated conservatively unless associated with injuries to the anterior cruciate ligament or posterior cruciate ligament. This article outlines rehabilitation programs for conservative treatment of MCL injuries, and postoperative programs after anterior cruciate ligament or multiple ligament reconstruction. In addition, the use of functional and prophylactic bracing for injuries of the MCL is reviewed. PMID- 17135957 TI - Biodegradable materials in arthroscopy. AB - The use of biodegradable materials as implants has revolutionized the way medicine is practiced today. This review provides a general description of salient biodegradable polymeric materials currently used in arthroscopy. These materials include polyglycolic acid, self-reinforced polyglycolic acid, poly-L lactic acid, self-reinforced polylactic-L-acid, poly-D-L-lactic acid, copolymer of poly-D-L-lactic acid polyglycolic acid, and polyglyconate. The mechanical strength, degradation properties, and widespread use of these materials, especially in the knee and shoulder, are discussed individually. Also discussed are the relatively few complications that are related to these materials' arthroscopic use. Future directions in biodegradable materials, including smart polymers, are also considered. In the future, novel techniques to identify the ideal polymer for a particular application will need to be developed to minimize the risk for implant complications. PMID- 17135958 TI - Biomedical devices in meniscal repair. AB - Preservation of the menisci is increasingly emphasized in orthopedic care of the knee. Techniques are rapidly emerging that allow easier accomplishment of this goal. In particular, the development of all-inside arthroscopic meniscal repair devices has facilitated more ready repair. A number of laboratory and clinical studies have examined these devices and comparisons with more traditional techniques have been made. Numerous reports of potential complications from these new technologies have also been described in the literature. This review covers the current options for treatment of meniscal repair, laboratory and clinical data, and also potential complications. PMID- 17135959 TI - Meniscal repair with fibrocartilage engineering. AB - Injuries to the knee meniscus, particularly those in the avascular region, pose a complex problem and a possible solution is tissue engineering of a replacement tissue. Tissue engineering of the meniscus involves scaffold selection, addition of cells, and stimulation of the construct to synthesize, maintain, or enhance matrix production. An acellular collagen implant is currently in clinical trials and there are promising results with other scaffolds, composed of both polymeric and natural materials. The addition of cells to these constructs may promote good matrix production in vitro, but has been studied in a limited manner in animal studies. Cell sources ranging from fibroblasts to stem cells could be used to overcome challenges in cell procurement, expansion, and synthetic capacity currently encountered in studies with fibrochondrocytes. Manipulation of construct culture with exogenous growth factors and mechanical stimulation will also likely play a role in these strategies. PMID- 17135960 TI - Clinical applications of bioactive factors in sports medicine: current concepts and future trends. AB - The ability to biologically manipulate musculoskeletal healing and augment bone and soft tissue repair and regeneration holds great promise. Advances in the basic science study and clinical application of bioactive proteins and growth factors continues to evolve. Improvement in the surgical resurfacing of articular cartilage defects and tendon and ligament repair through the addition of bioactive polypeptides is currently underway. The purpose of this article is to review the present array of biologically active materials that may be clinically applicable in sports medicine and arthroscopy. Mechanisms for biologic augmentation of tissue repair and regeneration will be discussed. Current limitations and future considerations will be reviewed particularly as they relate to practical clinical approaches. PMID- 17135961 TI - Articular cartilage: injury pathways and treatment options. AB - Articular cartilage injury and degeneration is a frequent occurrence in synovial joints. Treatment of these articular cartilage lesions are a challenge because this tissue is incapable of quality repair and/or regeneration to its native state. Nonoperative treatments endeavor to control symptoms, and include anti inflammatory medication, viscosupplementation, bracing, orthotics, and activity modification. Techniques to stimulate the intrinsic repair (fibrocartilage) process include drilling, abrasion, and microfracture of the subchondral bone. Currently, the clinical biologic approaches to treat cartilage defects include autologous chondrocyte implantation, periosteal transfer, and osteochondral autograft or allograft transplantation. Newer strategies employing tissue engineering being studied involve the use of combinations of progenitor cells, bioactive factors, and matrices, and the use of focal synthetic devices. Many new and innovative treatments are being explored in this exciting field. However, there is a paucity of prospective, randomized controlled clinical trials that have compared the various techniques, treatment options, indications and efficacy. PMID- 17135962 TI - Viscosupplementation therapy for osteoarthritis. AB - Intra-articular viscosupplementation with hyaluronic acid has become an increasingly accepted therapeutic alternative in the symptomatic management of osteoarthritis. Basic science research has documented numerous potential actions of exogenous hyaluronic acid upon the diseased joint. A substantial aggregate of clinical data suggests that viscosupplementation is clinically safe and suggests some therapeutic efficacy with regard to pain relief. However, a robust placebo effect and industry bias have obscured these outcomes considerably. Although recent attention has focused on the disease-modifying potential of this treatment option, definitive evidence is lacking. This paper will review the development, experience, indications, and clinical outcomes of viscosupplementation therapy in patients with osteoarthritis. PMID- 17135963 TI - Bone grafting in arthroscopy and sports medicine. AB - Bone graft substitutes include autografts, allografts, xenografts, and synthetics. Although autograft is still the gold standard, limited supply and donor morbidity must be considered. Allograft can vary in its bone-inductive qualities and may be processed into various shapes and constructs. Although allografts provide an osteoconductive matrix with some osteoinductivity, only limited anatomic constructs can be provided. Xenografts are abundant in supply, yet their shape and construct dimensions are restricted and xenograft properties are less than ideal due to the processing required to render the material nonimmunogenic. To achieve optimal bone graft properties, researchers are developing new materials with the goal of designing synthetics as close to autograft as possible. The advantages and disadvantages of all of these bone graft materials will be reviewed with emphasis on their relevance and applicability for sports medicine procedures. PMID- 17135964 TI - Biodegradable composite implants. AB - Biodegradable implants are established in the field of operative sports medicine. So-called composite implants are increasingly used. The idea to improve biocompatibility and osteoconductivity of biodegradable polymers by modifying them with a ceramic component, has led to the development of these composite implants. Today, an increasing variety of composite implants are available. They have substantially different material characteristics, which depend on matrix polymer choice and the additional ceramic. The material characteristics influence the mechanical properties, in vivo degradation, the osseous replacement and the host-tissue response. It is important to understand their biologic base for a better knowledge of the advantages and risks associated with using composite implants in the field of operative sports medicine. The purpose of this review is to focus on current developments in biodegradable composite implants and their biologic base. PMID- 17135965 TI - New sutures and suture anchors in sports medicine. AB - Arthroscopic surgery requires appropriate surgical implants for effective fixation of tendons and ligaments to bone. Biodegradable suture anchors are being used with increasing frequency for various procedures in sports medicine. As companions to these biodegradable suture anchors, new sutures have been developed which possess greater strength and different material properties from the conventional braided polyester suture. Biodegradable polymers currently found in sutures and suture anchors include poly-L-lactic acid, poly-D, L lactic acid, polydioxanone, polyglycolic acid and their copolymers. Suture anchors are now available preloaded with a choice of conventional braided polyester sutures or some version of ultrahigh molecular weight polyethylene ("super") sutures. Most new suture anchors come with 2 sutures. The manner in which these sutures are attached to the anchor varies and may consist of 2 separate eyelets or 2 slots either parallel to one another or at different angles to one another. Some anchors have a very large single eyelet that allows for 2 or more sutures. PMID- 17135966 TI - Tendon graft substitutes-rotator cuff patches. AB - Over the past few years, many biologic patches have been developed to augment repairs of large or complex tendon tears. These patches include both allograft and xenografts. Regardless of their origins, these products are primarily composed of purified type I collagen. Many factors should be considered when choosing an augmentation patch including tissue origin, graft processing, cross linking, clinical experience, and physical properties. The purpose of this article is to familiarize the sports medicine community with several tendon augmentation grafts: GraftJacket (Wright Medical Technology, Arlington, TN), CuffPatch (Organogenesis, Canton, MA, licensed to Arthrotek, Warsaw, IN), Restore (Depuy, Warsaw, IN), Zimmer Collagen Repair (Permacol) patch (Tissue Science Laboratories Covington, GA, licensed to Zimmer, Warsaw, IN), TissueMend (TEI Biosciences, Boston, MA, licensed to Stryker Howmedica Osteonics, Kalamazoo, MI), OrthoADAPT (Pegasus Biologics, Irvine, CA), and BioBlanket (Kensey Nash, Exton, PA). PMID- 17135968 TI - Treating the initial anterior shoulder dislocation--an evidence-based medicine approach. AB - This manuscript presents the best available evidence to answer questions regarding the treatment of the patient with an initial anterior shoulder dislocation. The highest levels of evidence available offer the following conclusions: (1) of the many methods to reduce the dislocated shoulder, little data exist to identify the best method. Recommendations are based on low levels of evidence (levels 4 and 5). (2) Premedication with intra-articular lidocaine has fewer complications and requires a shorter time in the emergency room than intravenous sedation with no detectable differences in reduction success rates (level 1). (3) Postreduction immobilization in external rotation may reduce recurrence (level 2), but immobilization in internal rotation does not (level 1). (4) Arthroscopic surgery significantly reduces recurrence compared to a nonoperative approach (level 1), and (5) there are limited data on features that would allow a safe return to play. Expert opinion suggests that return is allowed when motion and strength are nearly normal, and the athletes can engage in sport specific activities, however, the athlete is at risk for recurrence while playing (levels 4 and 5). PMID- 17135969 TI - Sideline management of sport-related concussions. AB - Concussions remain one of the most troublesome injuries sports physicians face. Studies suggest recovery takes hours to weeks, but at what point is the concussed brain no longer at increased risk for reinjury is unknown. Physicians must be alert to the symptoms of concussion and be familiar with the available tools to assess neurocognitive dysfunction. Prospectively validated signs and symptoms include amnesia, loss of consciousness, headache, dizziness, blurred vision, attention deficit, memory, postural instability, and nausea. A player with any signs or symptoms of a concussion should not be allowed to return to the current game or practice and should be monitored closely for deterioration of symptoms. Return-to-play should be individually based and proceed in a step-wise manner. The ongoing risk-benefit analysis of return-to-play must currently be based on experience, corollary data from traumatic brain injuries in animals and humans, and limited prospective data with sports-related concussions. PMID- 17135970 TI - Isolated posterior cruciate ligament injuries of the knee. AB - Isolated injuries of the posterior cruciate ligament (PCL) are rare in comparison to other ligamentous knee injuries, resulting in a lack of evidence-based literature regarding their treatment. Although consensus exists regarding the nonoperative management of asymptomatic grade I injuries, the treatment of high grade symptomatic PCL insufficiency remains a matter of debate. A variety of reconstructive procedures have been advocated on the basis of biomechanical data, however, the clinical benefit of these anatomic reconstructive techniques have yet to be conclusively proven in randomized trials. This article attempts to provide the treating physician with a concise overview of the etiology and diagnosis of isolated PCL injuries as well as an objective review of contemporary surgical treatment options and outcomes as reported in the current literature. PMID- 17135971 TI - Posterolateral corner injuries of the knee: anatomy, diagnosis, and treatment. AB - Injuries to the posterolateral corner of the knee continue to be a complex problem for orthopedic surgeons. Early recognition and treatment are important factors in the patient's long-term outcome. To properly treat these patients, the surgeon must have a clear understanding of the anatomic relationships amongst the structures in the posterolateral knee. This knowledge combined with a thorough physical examination and imaging studies, allows the surgeon to make the correct diagnosis and devise an appropriate treatment plan. This article will discuss the anatomy, diagnosis, and treatment options to improve the surgeon's understanding of posterolateral knee injuries. The senior author's technique for anatomic reconstruction of the posterolateral corner of the knee and the rehabilitation protocol are described. PMID- 17135972 TI - Evaluation and treatment of medial ulnar collateral ligament injuries in the throwing athlete. AB - Medial ulnar collateral ligament (UCL) injuries are common and are seen most frequently in baseball pitchers. Appropriate recognition, treatment, and rehabilitation are necessary to ensure the best chance for return to preinjury levels of participation. Participation in competitive sports may be disrupted for 6 months to 1 year when treated optimally. Abstinence from play may be prolonged when treatment is delayed or if conservative treatment fails; this delay carries significant consequences to the professional, collegiate, and high school athlete. The orthopedic literature is replete with recommendations for the care of these athletes. These recommendations are generally based on retrospective reviews. The purposes of this paper are 3-fold: to provide background knowledge on this injury, to synthesize the current knowledge on the diagnosis, treatment, and rehabilitation of athletes with medial UCL injuries, and lastly, to provide a treatment algorithm for athletes with UCL injuries. PMID- 17135973 TI - Evidence-based approach to treatment of acute traumatic syndesmosis (high ankle) sprains. AB - Ankle sprains in the athlete are one of the most common injuries, and syndesmosis or "high-ankle" sprains seem to being diagnosed at an increasing rate. As a result, there has been a heightened interest in recognizing and treating these difficult injuries on a timely basis, particularly in the athlete. Although the recognition and diagnosis of these injuries have improved, there still exists a paucity of information on optimal conservative and operative management. In this paper, a systematic review of the literature was conducted to provide an evidence based rationale in the diagnosis and treatment of syndesmosis (high ankle) sprains in athletes. It is obvious from the low level of evidence available in the literature on this topic that a great deal of work is needed before conclusive statements regarding the management of these injuries can be made with confidence. The current diagnostic tests are not very specific. Because this is a spectrum of injury, there is a lot of variability in the time lost from sport. It is clear that we need a much more definitive diagnostic process for this injury that allows us to predict the severity of the injury, time loss from sport, and the treatment required. PMID- 17135974 TI - Treatment of the acute traumatic acromioclavicular separation. AB - Injuries to the acromioclavicular joint occur commonly in athletes, especially those involved in contact sports. The majority of these injuries are type I and II acromioclavicular joint separations and are treated nonoperatively with rehabilitation. A rapid and full return to play is expected. Acute types IV, V, and VI are less common and operative intervention is recommended. The type III injury is more controversial and current trends are towards initial nonoperative management. Operative treatment is sought only when the athlete remains symptomatic with painful instability. However, some do support early intervention in the overhead athlete. The goal of operative intervention is to create a stiff and strong repair/reconstruction of the coracoclavicular ligaments while providing stability in all planes. This will allow early and more aggressive rehabilitation. Surgical treatment includes reconstruction of the coracoclavicular ligaments with an augmented coracoacromial ligament transfer and more recently tendon graft reconstructions. Biomechanical research supports an anatomic reconstruction of the ligaments to confer the most function and stability. PMID- 17135975 TI - Troublesome stress fractures of the foot and ankle. AB - Athletes engaged in repetitive activity with recent increases in training or competition are at risk for stress fractures of the foot and ankle. Physical and radiographic findings are often unremarkable and diagnosis may be difficult. Although many of these injuries may be managed symptomatically, certain types of stress fractures are at higher risk for delayed union, nonunion, or refracture without aggressive immobilization or surgical fixation. PMID- 17135978 TI - An analysis of surgical complications, morbidity, and cost calculation in patients undergoing multimodal treatment for operable oropharyngeal carcinoma. AB - OBJECTIVE/HYPOTHESIS: Tumor control and survival are considered the most important measures of treatment efficacy for patients with primary oropharyngeal squamous cell carcinoma. Furthermore, multimodal treatment protocols should be judged by their complication rates, morbidity, and therapy costs. STUDY DESIGN: The results of a combined approach of primary surgery and neck dissection with postoperative radio(chemo)therapy were analyzed in retrospective chart review. METHODS: Two hundred eleven patients' records were analyzed for surgical complications, therapeutic morbidity, and treatment costs. RESULTS: The rate of postoperative hemorrhage was 4.7%. We observed no fatal complications. Ten percent of our patients required nutrition through percutaneous endoscopic gastrostomy (PEG). Twelve percent of all patients required long-term tracheostomy. The rates of PEG and tracheostomy were significantly higher in patients operated by the transcervical approach. The costs for the combined approach ranged from 10,587 euros (13,377 dollars) to 24,531 euros (30,996 dollars). CONCLUSIONS: The presented multimodal approach provides a low rate of surgical complications and a tolerable morbidity. Considering the excellent oncologic results, this extensive and more cost-intensive multimodal approach is justified for patients with oropharyngeal cancer. PMID- 17135979 TI - Cochlear implantation in a patient with bilateral deep brain stimulators. AB - OBJECTIVE: We report the case of a patient successfully implanted with a Nucleus Contour cochlear implant after placement of a deep brain stimulator for Parkinson disease. METHODS: The authors conducted a case report and literature review. RESULTS: Successful hookup and mapping of the device was performed 1 month after implantation without evidence of aberrant activity of the deep brain stimulators. CONCLUSIONS: To our knowledge, this is the first reported case of successful implantation of both a cochlear implant and a deep brain stimulator in the same patient. We have outlined one approach to avoiding detrimental interactions between cochlear implant and deep brain stimulator devices. PMID- 17135980 TI - Endoscopic surgery for sinonasal invasive aspergillosis in bone marrow transplantation patients. AB - OBJECTIVES/HYPOTHESIS: Sinonasal invasive aspergillosis (IA) is an aggressive fungal infection with high mortality rates. It commonly develops in immunocompromised patients, often after bone marrow transplantation (BMT). Aggressive surgical debridement by an external approach has been considered a central element of treatment. We describe our experience in endoscopic management of IA in BMT patients in a retrospective study. METHODS: Charts of BMT patients with IA in the past 5 years were reviewed. Demographic data, primary disease, comorbidities, signs and symptoms, blood test results, preparation for surgery, surgical technique, and outcome were recorded. RESULTS: Fourteen BMT patients, age ranging from 3 to 56 years, had sinonasal IA. The primary disease was acute myelogenous leukemia in 6, acute lymphoblastic leukemia in 3, chronic myeloblastic leukemia in one, severe combined immunodeficiency disease in 2, and myelodysplastic syndrome in 2 patients. Diagnosis was made by physical examination, biopsy, culture, and computed tomography scan. Treatment, including aggressive endoscopic debridement, a systemic antifungal medication, and local irrigations of amphotericin-B enabled eradication of IA in all patients. Seven patients required two or more operations. None required orbital exenteration or craniotomy. Six patients died of the primary illness or of comorbidities with no evidence of residual disease. Eight patients are alive. CONCLUSION: Early detection of IA in BMT patients enables aggressive treatment before the disease spreads into the orbit or brain. Proper preoperative preparation facilitates safe endoscopic surgery in patients with severe bleeding tendencies. Although sinonasal IA is lethal, endoscopic surgery is feasible and efficient, enabling excellent local control. PMID- 17135981 TI - Aortic arch laceration: A lethal complication after percutaneous tracheostomy. AB - Percutaneous dilatational tracheostomy (PDT) is not a new technique. However, it was not until Ciaglia introduced a simple technique using a needle, catheter, and guidewire that it became a possible alternative to open surgical tracheotomy. PDT is not performed blindly but with the assistance of fiberoptic endoscopy. However, even with fiberoptic endoscopic control, PDT can give rise to serious complications similar to those encountered with open surgical tracheostomy. We present an uncommon complication of percutaneous tracheostomy, a case of aortic arch laceration. We discuss the anatomy of the pretracheal space and how to avoid such a lethal complication. PMID- 17135982 TI - Pneumococcal peptidoglycan-polysaccharides induce the expression of interleukin-8 in airway epithelial cells by way of nuclear factor-kappaB, nuclear factor interleukin-6, or activation protein-1 dependent mechanisms. AB - Cell envelope compounds of bacteria trigger immune and inflammatory reactions by way of chemokines/cytokines. In this study, we demonstrated that pneumococcal peptidoglycan-polysaccharides (PGPS) induced the production of interleukin (IL)-8 by way of nuclear factor (NF)-kappaB, nuclear factor interleukin (NF-IL)6, and activation protein (AP)-1 dependent mechanisms in the human bronchial epithelial cells (NL-20) in a dose- and time-dependent manner in vitro, and the mutation of either the NF-kappaB, NF-IL6, or AP-1 binding sites in the promoter of IL-8 abrogated the IL-8 transcriptional activity. In a similar way, lipopolysaccharides induced the promoter activation of IL-8 in NL-20. However, the PGPS-induced IL-8 promoter activation in rodent middle ear epithelial cells required NF-kappaB and NF-IL6 but not AP-1. PMID- 17135984 TI - The Treatment for Adolescents with Depression Study (TADS): methods and message at 12 weeks. AB - Funded by the National Institute of Mental Health, the Treatment for Adolescents With Depression Study (TADS) is intended to evaluate the short-term (12 weeks) and longer-term (36 weeks) effectiveness of four treatments for adolescents with DSM-IV major depressive disorder: clinical management with fluoxetine (FLX), cognitive-behavioral therapy (CBT), FLX and CBT combined (COMB), and clinical management with placebo (PBO). We previously reported that COMB and FLX were more effective in reducing depression than CBT or PBO after 12 weeks of acute treatment. In this special section of the Journal, separate articles extend these findings to the impact of TADS treatments on remission, speed of response, function and quality of life, predictors of outcome, and safety during the first 12 weeks of treatment. To set the stage for the special section, we briefly review the rationale, design, and methods of the TADS; describe the TADS sample to which the TADS findings generalize; using all of the currently available data, summarize the intent-to-treat outcomes across multiple endpoints at 12 weeks; and consider the public health value of the TADS findings in the context of design decisions and methodological limitations of the TADS, including some that may have advantaged the combined treatment condition. Reflecting the ordering of effect sizes at week 12--COMB (0.98) > FLX (0.68) > CBT (-0.03)--combined treatment proved superior to PBO on 15 of 16 endpoints, to CBT on 14 of 16 endpoints, and to FLX on 8 of 16 endpoints, whereas FLX was superior to CBT on 8 of 14 and to PBO on 7 of 16 measures. CBT did not differ from PBO on any measure. Despite the fact that suicidality improved markedly across all of the treatment conditions, suicidal events were twice as common in patients treated with FLX alone than with COMB or CBT alone, perhaps indicating that CBT protects against suicidal events. Thus, combined treatment appears to accelerate recovery relative to CBT and, for some outcomes, FLX alone, while minimizing the risk of suicidality relative to FLX alone. Taking benefit and risk into account, we conclude that the combination of FLX and CBT appears superior to either monotherapy as a treatment for moderate to severe major depressive disorder in adolescents. PMID- 17135985 TI - Remission and residual symptoms after short-term treatment in the Treatment of Adolescents with Depression Study (TADS). AB - OBJECTIVE: To ascertain remission rates in depressed youth participating in the Treatment for Adolescents With Depression Study (TADS), a multisite clinical trial that randomized 439 adolescents with major depressive disorder (MDD) to a 12-week treatment of fluoxetine (FLX), cognitive-behavioral therapy (CBT), their combination (COMB), or clinical management with pill placebo (PBO). METHOD: Using an end-of-treatment Children's Depression Rating Scale-Revised (CDRS-R) total score of 28 or below as the criterion for remission, rates of remission were examined with logistic regression, controlling for site. Loss of MDD diagnosis and residual symptoms in responders (defined as Clinical Global Impressions Improvement (CGI-I) score of 1 (very much improved) or 2 (much improved) were also examined across treatment groups. RESULTS: After 12 weeks of treatment, 102 (23%) of 439 youths had reached remission. The remission rate was significantly higher in the COMB group (37%) relative to the other treatment groups (FLX, 23%; CBT, 16%; PBO, 17%), with odds ratios of 2.1 for COMB versus FLX, 3.3 for COMB versus CBT, and 3.0 for COMB versus PBO. In addition, 71% of subjects across treatment groups no longer met criteria for MDD at the end of acute treatment. Fifty percent of the youths who responded by CGI-I criteria continued to have residual symptoms, such as sleep or mood disturbances, fatigue, and poor concentration. CONCLUSIONS: The combination of FLX and CBT was superior to both monotherapy and PBO in terms of remission rates, but overall rates of remission remain low and residual symptoms are common at the end of 12 weeks of treatment. PMID- 17135986 TI - Acute time to response in the Treatment for Adolescents with Depression Study (TADS). AB - OBJECTIVE: To examine the time to response for both pharmacotherapy and psychotherapy in the Treatment for Adolescents with Depression Study (TADS). METHOD: Adolescents (N = 439, ages 12 to 17 years) with major depressive disorder were randomized to fluoxetine (FLX), cognitive-behavioral therapy (CBT), their combination (COMB), or pill placebo (PBO). Defining response as very much improved or much improved on the Clinical Global Impression-Improvement Scale (CGI-I), survival analyses using Cox proportional hazards models, and Kaplan Meier curves were conducted to evaluate time to first response and time to stable response for subjects receiving pharmacotherapy (COMB, FLX, PBO) as well as for subjects receiving CBT (COMB, CBT). Direct comparisons between pharmacotherapy and CBT were not made because of differences in visit schedules. RESULTS: Based on pharmacotherapist CGI-I scores, COMB and FLX showed faster onset of benefit than PBO on time to response and time to stable response (p < .001), and COMB was faster than FLX on time to stable response (p = .034). The probability of sustained early response was approximately threefold greater for COMB than PBO, twofold greater for FLX than PBO, and 1.5-fold greater for COMB than FLX. On the psychotherapist CGI-I scores, both first response and stable response occurred faster in COMB than CBT (p < .001), with a probability of sustained early response approximately threefold greater for COMB than CBT. CONCLUSIONS: In the acute treatment of depressed adolescents, FLX and COMB accelerate response relative to PBO, and COMB accelerates response relative to CBT alone. PMID- 17135987 TI - Functioning and quality of life in the Treatment for Adolescents with Depression Study (TADS). AB - OBJECTIVE: To test whether 12-week treatment of major depression improved the level of functioning, global health, and quality of life of adolescents. METHOD: The Treatment for Adolescents With Depression Study was a multisite, randomized clinical trial of fluoxetine, cognitive-behavioral therapy (CBT), their combination (COMB), or clinical management with placebo in 439 adolescents with major depression. Functioning was measured with the Children's Global Assessment Scale (CGAS), global health with the Health of the Nation Outcome Scales for Children and Adolescents (HoNOSCA), and quality of life with the Pediatric Quality of Life Enjoyment and Satisfaction Questionnaire (PQ-LES-Q). Random effects regression models were applied to the data. RESULTS: Compared with placebo, COMB was effective on the CGAS (p < .0001), HoNOSCA (p < .05), and PQ LES-Q (p < .001), whereas fluoxetine was superior to placebo on the CGAS only (p < .05). COMB was superior to fluoxetine on the CGAS (p < .05) and PQ-LES-Q (p = .001). Fluoxetine was superior to CBT on the CGAS (p < .01). CBT monotherapy was not statistically different from the placebo group on any of the measures assessed. Treatment effects were mediated by improvement in depressive symptoms measured on the Child Depression Rating Scale-Revised. CONCLUSIONS: The combination of fluoxetine and CBT was effective in improving functioning, global health, and quality of life in depressed adolescents. Fluoxetine monotherapy improved functioning. PMID- 17135988 TI - Predictors and moderators of acute outcome in the Treatment for Adolescents with Depression Study (TADS). AB - OBJECTIVE: To identify predictors and moderators of response to acute treatments among depressed adolescents (N = 439) randomly assigned to fluoxetine, cognitive behavioral therapy (CBT), both fluoxetine and CBT, or clinical management with pill placebo in the Treatment for Adolescents With Depression Study (TADS). METHOD: Potential baseline predictors and moderators were identified by a literature review. The outcome measure was a week 12 predicted score derived from the Children's Depression Rating Scale-Revised (CDRS-R). For each candidate moderator or predictor, a general linear model was conducted to examine main and interactive effects of treatment and the candidate variable on the CDRS-R predicted scores. RESULTS: Adolescents who were younger, less chronically depressed, higher functioning, and less hopeless with less suicidal ideation, fewer melancholic features or comorbid diagnoses, and greater expectations for improvement were more likely to benefit acutely than their counterparts. Combined treatment, under no condition less effective than monotherapy, was more effective than fluoxetine for mild to moderate depression and for depression with high levels of cognitive distortion, but not for severe depression or depression with low levels of cognitive distortion. Adolescents from high-income families were as likely to benefit from CBT alone as from combined treatment. CONCLUSIONS: Younger and less severely impaired adolescents are likely to respond better to acute treatment than older, more impaired, or multiply comorbid adolescents. Family income level, cognitive distortions, and severity of depression may help clinicians to choose among acute interventions, but combined treatment proved robust in the presence of moderators. PMID- 17135989 TI - Treatment for Adolescents with Depression Study (TADS): safety results. AB - OBJECTIVE: To compare the rates of physical, psychiatric, and suicide-related events in adolescents with MDD treated with fluoxetine alone (FLX), cognitive behavioral therapy (CBT), combination treatment (COMB), or placebo (PBO). METHOD: Safety assessments included adverse events (AEs) collected by spontaneous report, as well as systematic measures for specific physical and psychiatric symptoms. Suicidal ideation and suicidal behavior were systematically assessed by self- and clinician reports. Suicidal events were also reanalyzed by the Columbia Group and expert raters using the Columbia-Classification Algorithm for Suicidal Assessment used in the U.S. Food and Drug Administration reclassification effort. RESULTS: Depressed adolescents reported high rates of physical symptoms at baseline, which improved as depression improved. Sedation, insomnia, vomiting, and upper abdominal pain occurred in at least 2% of those treated with FLX and/or COMB and at twice the rate of placebo. The rate of psychiatric AEs was 11% in FLX, 5.6% in COMB, 4.5% in PBO, and 0.9% in CBT. Suicidal ideation improved overall, with greatest improvement in COMB. Twenty-four suicide-related events occurred during the 12-week period: 5 patients (4.7%) in COMB, 10 (9.2%) in FLX, 5 (4.5%) in CBT, and 3 (2.7%) in placebo. Statistically, only FLX had more suicide-related events than PBO (p =.0402, odds ratio (OR) = 3.7, 95% CI 1.00-63.7). Only five actual attempts occurred (2 COMB, 2 FLX, 1 CBT, 0 PBO). There were no suicide completions. CONCLUSIONS: Different methods for eliciting AEs produce different results. In general, as depression improves, physical complaints and suicidal ideation decrease in proportion to treatment benefit. In this study, psychiatric AEs and suicide-related events are more common in FLX-treated patients. COMB treatment may offer a more favorable safety profile than medication alone in adolescent depression. PMID- 17135990 TI - After TADS, can we measure up, catch up, and ante up? PMID- 17135991 TI - Glad for what TADS adds, but many TADS grads still sad. PMID- 17135992 TI - A pilot study of modified cognitive-behavioral therapy for childhood traumatic grief (CBT-CTG). AB - OBJECTIVE: This pilot study evaluated outcomes for a modified 12-session protocol of cognitive-behavioral therapy for childhood traumatic grief (CBT-CTG) conducted between March 2004 and October 2005. CTG is an emerging condition characterized by a combination of posttraumatic stress and unresolved grief symptoms. This two module treatment model consisting of sequential trauma- and grief-focused components was shortened from a previously presented 16-session protocol. METHOD: Thirty-nine children ages 6 to 17 years old with CTG and their parents received the modified 12-session protocol of CBT-CTG. CTG and posttraumatic stress disorder (PTSD) symptoms were assessed at pretreatment, after the trauma-focused module, and after the grief-focused module (at posttreatment). Child depression, anxiety, and behavioral symptoms, as well as parental depression and PTSD symptoms, were assessed at pre- and posttreatment. RESULTS: Children reported significant improvement in CTG, PTSD, depression, and anxiety, and parents reported significant improvement in children's PTSD, internalizing and total behavior problems, and their personal PTSD symptoms. Although PTSD significantly improved only during the trauma-focused module of treatment, CTG improved significantly during both trauma- and grief-focused modules of treatment. Child satisfaction and parent satisfaction for this treatment protocol were also high. CONCLUSIONS: These findings suggest that the shortened CBT-CTG protocol, which is similar in the number of sessions to what many community child bereavement programs offer, may be acceptable and efficacious for this population. The CBT CTG model requires further evaluation in randomized, controlled treatment trials. PMID- 17135993 TI - A follow-up study of a multisite, randomized, controlled trial for children with sexual abuse-related PTSD symptoms. AB - OBJECTIVE: To ascertain whether the differential responses that previously have been found between trauma-focused, cognitive-behavioral therapy (TF-CBT), and child-centered therapy (CCT) for treating posttraumatic stress disorder (PTSD) and related problems in children who had been sexually abused would persist following treatment and to examine potential predictors of treatment outcome. METHOD: A total of 183 children 8 to 14 years old and their primary caregivers were assessed 6 and 12 months after their posttreatment evaluations. RESULTS: Mixed-model repeated analyses of covariance found that children treated with TF CBT had significantly fewer symptoms of PTSD and described less shame than the children who had been treated with CCT at both 6 and 12 months. The caregivers who had been treated with TF-CBT also continued to report less severe abuse specific distress during the follow-up period than those who had been treated with CCT. Multiple traumas and higher levels of depression at pretreatment were positively related to the total number of PTSD symptoms at posttreatment for children assigned to the CCT condition only. CONCLUSIONS: Children and caregivers assigned to TF-CBT continued to have fewer symptoms of PTSD, feelings of shame, and abuse-specific parental distress at 6- and 12-month assessments as compared to participants assigned to CCT. PMID- 17135994 TI - Parent-child agreement regarding children's acute stress: the role of parent acute stress reactions. AB - OBJECTIVE: We examined parent-child agreement regarding child acute stress disorder (ASD) and the relationship between parent ASD symptoms and parent ratings of child ASD. METHOD: Parent-child dyads (N = 219; child age 8-17 years) were assessed within 1 month of child injury. Parent-child agreement was examined regarding child ASD presence, severity, and specific symptoms. Relationships among parent ASD and parent- and child-reported child ASD were examined using regression analysis and generalized estimating equations (GEE). RESULTS: Parent child agreement was low for presence of child ASD (kappa = 0.22) and for individual symptoms. Parent and child ratings of child ASD severity were moderately correlated (r = 0.35). Parent ASD was independently associated with parent-rated child ASD, after accounting for child self-rating (beta =.65). Generalized estimating equations indicated that parents with ASD overestimated child ASD and parents without ASD underestimated child ASD, compared to the child's self-rating. CONCLUSIONS: Parents' own responses to a potentially traumatic event appear to influence their assessment of child symptoms. Clinicians should obtain child self-report of ASD whenever possible and take parent symptoms into account when interpreting parent reports. Helping parents to assess a child's needs following a potentially traumatic event may be a relevant target for clinical attention. PMID- 17135995 TI - Family relational factors in pediatric depression and asthma: pathways of effect. AB - OBJECTIVE: This study tested a multilevel biobehavioral family model proposing that negative family emotional climate contributes to child depressive symptoms, which in turn contribute to asthma disease severity. Parent-child relational insecurity is proposed as a mediator. METHOD: Children with asthma (N = 112; ages 7-18; 55% male) reported relational security, anxiety, and depressive symptoms. Parent(s) reported demographics, asthma history and symptoms, and family emotional expression. Asthma diagnosis was confirmed by medical history provided by parent and child together, clinical evaluation, pulmonary function tests, and methacholine challenge, with disease severity categorized by National Heart, Lung, and Blood Institute guidelines. Medication adherence was measured prospectively. RESULTS: Path analysis indicated a good fit of data to the hypothesized model (chi2 = 0.072, p =.97, normal fit index = 0.998, root mean square error of approximation = 0.000). Negative family emotional climate predicted child depressive symptoms (beta =.21, p < .04), which predicted asthma disease severity (beta =.35, p < .001), with relational insecurity a partial mediator (beta = -.23, p < .05, beta =.46, p < .001, respectively). Depression was associated with disease severity even after controlling for adherence (r p = 0.38, p < .05). CONCLUSION: Findings are consistent with the proposed family model, suggesting the clinical importance of assessing and intervening in these specific family relational processes when treating children with depression and asthma. PMID- 17135996 TI - Steady-state clinical pharmacokinetics of bupropion extended-release in youths. AB - OBJECTIVE: To examine in children and adolescents the 24-hour, steady-state clinical pharmacokinetics of an extended-release (XL) formulation of bupropion (Wellbutrin XL). METHOD: Subjects were six male and four female patients (ages 11.5-16.2 years) prescribed bupropion XL in morning daily doses of either 150 mg (n = 5) or 300 mg (n = 5) for at least 14 days. During an overnight hospitalization, subjects had serial blood draws every 1.5 to 3 hours from an intravenous port to measure plasma levels of bupropion and its metabolites. Pharmacokinetic variables were determined by noncompartmental analysis for bupropion and exponential analyses for metabolites. RESULTS: Bupropion and metabolites demonstrated linear pharmacokinetics. Bupropion's mean maximum concentration (Cmax) was lower (p = .021) and its mean time to Cmax longer (p = .057) in the current sample on bupropion XL relative to a previously studied sample of youths on bupropion sustained-release (Wellbutrin SR). Mean 24-hour area under the curve ratios of metabolites to bupropion ranged from 1.0 for erythrohydrobupropion to 16.4 for hydroxybupropion. CONCLUSIONS: Once-daily dosing is justified in youths prescribed bupropion XL. The active metabolite hydroxybupropion probably has key pharmacodynamic effects, given its higher and more sustained levels relative to the other metabolites or to bupropion. PMID- 17135997 TI - Early father involvement moderates biobehavioral susceptibility to mental health problems in middle childhood. AB - OBJECTIVE: To study how early father involvement and children's biobehavioral sensitivity to social contexts interactively predict mental health symptoms in middle childhood. METHOD: Fathers' involvement in infant care and maternal symptoms of depression were prospectively ascertained in a community-based study of child health and development in Madison and Milwaukee, WI. In a subsample of 120 children, behavioral, autonomic, and adrenocortical reactivity to standardized challenges were measured as indicators of biobehavioral sensitivity to social context during a 4-hour home assessment in 1998, when the children were 7 years of age. Mental health symptoms were evaluated at age 9 years using parent, child, and teacher reports. RESULTS: Early father involvement and children's biobehavioral sensitivity to context significantly and interactively predicted symptom severity. Among children experiencing low father involvement in infancy, behavioral, autonomic, and adrenocortical reactivity became risk factors for later mental health symptoms. The highest symptom severity scores were found for children with high autonomic reactivity that, as infants, had experienced low father involvement and mothers with symptoms of depression. CONCLUSIONS: Among children experiencing minimal paternal caretaking in infancy, heightened biobehavioral sensitivity to social contexts may be an important predisposing factor for the emergence of mental health symptoms in middle childhood. Such predispositions may be exacerbated by the presence of maternal depression. PMID- 17135998 TI - Geometrically evident: framing studies using the Graphic Appraisal Tool for Epidemiology (GATE). PMID- 17135999 TI - Pharmacological management of a youth with ADHD and a seizure disorder. PMID- 17136001 TI - Nitric oxide's pulsatile release in lobster heart and its regulation by opiate signaling: pesticide interference. AB - BACKGROUND: Data is emerging in the human and invertebrate literature demonstrating that mu opiate receptors and morphine are present in cardiovascular tissues in diverse animals, including human tissues, where they may be exerting a cardioregulatory role via stimulation of constitutive nitric oxide (NO) production. MATERIAL/METHODS: NO release from lobster heart was evaluated without stimulation and after morphine exposure using a real-time NO-specific amperometric probe. In addition, real time NO release was evaluated after treatment with low doses of widely used pesticides (e.g., pyrethroids). Real time RT-PCR was used to investigate the presence of mu opiate receptor subtypes in lobster heart. RESULTS: Basal NO release occurs in lobster heart at the nanomolar level. Morphine enhanced this level of release; naloxone (an opiate antagonist) blocked it, as did exposure to the NO synthase inhibitor L-NAME. In addition, treatment with the pyrethroids, permethrin and resmethrin, abrogated constitutive NO release from lobster heart. Finally, by way of real time RT-PCR we were able to demonstrate the presence of the micro(3) opiate receptor subtype in lobster heart. CONCLUSIONS: Rhythmic NO bursts appear to be involved in normal cardiac muscle activity in Homarus americanus. Lobster heart contains morphinergic signaling components capable of slowing down its beating rate via NO production. In addition, compounds such as pyrethroid pesticides may alter normal cardiac activity by interfering with constitutive NO production and thus, depressing basal NO levels. This may ultimately make these animals more susceptible to environmental assaults/toxins. PMID- 17136002 TI - PolicosanolPlus and Neuroprevin ameliorate pesticide-mediated inhibition of neurite outgrowth and neurite degeneration. AB - BACKGROUND: Pesticide exposure is a recognized risk factor for neurodegenerative diseases. Recently, bifenthrin, a pyrethroid pesticide, was shown to inhibit the formation of neurites and cause neurite retraction, raising concern that these newer and less toxic pesticides may also contribute to neurodegenerative diseases. PolicosanolPlus and Neuroprevin are nutraceutical supplements which promote the survival of neurites in neuronal cell cultures. Here we determine if PolicosanolPlus and Neuroprevin can ameliorate the neurodegenerative effects of bifenthrin. MATERIAL/METHODS: PC12 cells were treated with NGF, bifenthrin, PolicosanolPlus and Neuroprevin in various combinations and the formation of neurites was assessed microscopically at times ranging from 12 to 72 hours post treatment. Bifenthrin was also withheld at the time of NGF, PolicosanolPlus and Neuroprevin treatment and added after neurite formed to assess neurite retraction. RESULTS: Bifenthrin (1 x 10(-6) M) inhibits neurite outgrowth, in the absence of cell death, by more than 50% at 12 hours and by more than 80% at 72 hours. With addition of PolicosanolPlus and/or Neuroprevin at the time of cell seeding, bifenthrin does not inhibit neurite outgrowth. Addition of bifenthrin to differentiated cells results in a retraction of 90% of neurites, while those with PolicosanolPlus and Neuroprevin show no significant retraction of neurites. CONCLUSIONS: The pesticide, bifenthrin, inhibits neurite formation and causes neurite retraction. PolicosanolPlus and Neuroprevin are nutraceutical supplements which ameliorate the effects of bifenthrin on neurite outgrowth and retraction. Dietary supplementation with PolicosanolPlus and Neuroprevin may protect against developmental and long-term neurodegenerative events that result from exposure to pesticides. PMID- 17136003 TI - p53 gene gets altered by various mechanisms: studies in childhood sarcomas and retinoblastoma. AB - BACKGROUND: Somatic and constitutional mutation screening of p53 in childhood sarcomas and retinoblastoma was investigated by a multitechnical approach to evaluate its role in the development/progression by somatic mutation events and/or genetic predisposition. MATERIAL/METHODS: The studies were carried out on a cohort of 100 sarcoma cases, i.e. Ewing's sarcoma (n=44), osteosarcoma (n=36), and rhabdomyosarcoma (n= 20), and on 50 retinoblastoma (Rb) cases. RESULTS: Constitutional allelic deletion was found by FISH in 4% of sarcoma cases. Overall, 20% of sarcoma tumors showed p53 rearrangement by PCR/SSCP and Southern blot. Allelic deletion of p53 was detected in 78% of sarcoma and 55% of Rb tumors. p53 protein expression was detected by immunohistochemistry in 20% of sarcoma tumors. CONCLUSIONS: This study for the first time provided evidence of p53 alteration through allelic deletion that are common primary somatic mutation events which occur irrespective of grade and stage and are hence probably associated with an early phase of tumorigenesis and/or tumor progression. The studies also explored the occurrence of de novo constitutional deletion of p53 in sporadic childhood sarcomas. This study in retinoblastoma provided evidence for the synergistic role of RB1 and p53, probably essential for the full-blown development of malignancy. PMID- 17136004 TI - An adherence typology: coping, quality of life, and physical symptoms of people living with HIV/AIDS and their adherence to antiretroviral treatment. AB - BACKGROUND: Research has found that patient adherence to antiretroviral therapy is crucial to treatment success, but this research did not investigate the patient's viewpoint. This study examined relationships between types of adherence and coping, psychosocial factors, quality of life (QoL), and physical symptoms from the perspective of people living with HIV/AIDS. MATERIAL/METHODS: The quantitative study involved 100 HIV-positive participants. Questionnaires comprised the Trier Scales on Coping with Physical Illness, Medical-Outcomes Study HIV Health-Survey QoL, Social Factors of Antiretroviral Therapy, and the HIV/AIDS Physical Symptom-Checklist. A sub-sample of 41 participants underwent semi-standardized interviews eliciting the type of adherence. Grounded Theory was the method of qualitative analyses. RESULTS: Maladaptive coping (rumination) related to poor mental health (p<0.001), concealing the HIV-infection (p<0.01), and being treatment-naive (p<0.01). Spiritual coping was more likely in women (p<0.001). Overall, QoL was worse in participants with more physical symptoms (p<0.001) and in those seeking mental health care (p<0.001). Working and maintaining a regular daily routine were associated with better adherence (p<0.05). Four adherence types were identified: 'Traditional Adherence' (with indifferent, faithful, and anxious subtypes), 'Traditional Non-Adherence', 'Critical Adherence', and 'Critical Non-Adherence'. The traditional types underscored a paternalistic medical model, while critical types emphasized 'autonomous patients'. Critical types were less frequent (39%), although superior to traditional types (p<0.001) in internal locus of control, optimal social support, and adaptive coping. CONCLUSIONS: Critical adherence is superior to traditional adherence with respect to physical and psychosocial factors. Strategies to improve adherence should therefore target empowerment and autonomy rather than patient obedience. PMID- 17136005 TI - Free thyroxine index and risk of stroke: results from the National Health and Nutrition Examination Survey Follow-up Study. AB - BACKGROUND: The long-term risks of stroke associated with hypothyroidism and hyperthyroidism is unknown. We evaluated the long-term risk of stroke with hypothyroidism and hyperthyroidism compared with euthyroid status. MATERIAL/METHODS: We used the 20-year follow-up data for adults aged 25 to 74 years who participated in the First National Health and Nutrition Examination Survey Follow-up Study. Hypothyroidism or hyperthyroidism was diagnosed by free thyroxine index measurements at baseline evaluation and incident events were determined from hospital records and death certificates during follow-up. Relative risk (RR) of stroke, ischemic stroke, and intracerebral hemorrhage for each category of thyroid function were determined by Cox proportional hazards analysis after adjustment for potential confounding variables. RESULTS: A total of 5,269 participants (mean age 48+/-14 years; 2,379 men) were evaluated. Hypothyroidism and hyperthyroidism were diagnosed in 493 (9.4%) and 34 (0.7%) participants, respectively. After adjustment for covariates, a significantly higher RR for all strokes and for ischemic stroke was observed in participants with hypothyroidism (RR 1.6; 95% confidence interval (CI), 1.0-2.6 and RR 1.6, 95% CI, 1.0-2.7, respectively). Hyperthyroidism was not associated with an increased risk for stroke. CONCLUSIONS: An increased risk for stroke (particularly ischemic stroke) was observed in patients with hypothyroidism. PMID- 17136006 TI - Gait analysis in osteoarthritis of the hip. AB - BACKGROUND: Several characteristic groups of changes can be observed during full instrumental evaluation of gait in patients with osteoarthritis of the hip. However, this procedure is expensive and causes patient discomfort. We attempted to determine the most clinically useful parameters of gait changes in patients with osteoarthritis of the hip measured by pedobarography, a simple and non invasive technique. MATERIAL/METHODS: 30 patients of both genders, 51-78 years of age, were included in the study. Each patient was tested for static and dynamic gait parameters using an EMED SF-4 force platform (Novel, Germany), with a grid of force transducers registering the pattern of forces exerted by the sole of the foot on ground contact. RESULTS: Significant differences were observed in step length. Small but statistically significant differences were observed for total ground contact area between the results obtained for the affected and helthy leg. Maximum ground reaction measured for the whole foot and ground contact time was not significantly different between the limbs. Statistically significant differences were observed for maximum pressure and area under the pressure/contact time curve. In evaluating the gaitline (a line representing the maximum dynamic force from initial contact to push-off), 25 of 30 patients had an abnormal result for the affected leg, whereas the healthy leg was abnormal in 16 of 30 patients. CONCLUSIONS: Gait abnormalities in patients with hip osteoarthritis are mainly observable in asymmetry of weight-bearing and asymmetry of step length. Pedobarography is a clinically suitable diagnostic tool. PMID- 17136007 TI - Does the inactivation of leukocytes in blood transfusions during and following liver transplantation by gamma-irradiation have an impact on rejection and infection rate? AB - BACKGROUND: Leukocytes transmitted in blood products exert a variety of immunological side-effects. Experience with bone marrow transplant recipients has shown that these can be induced even by very few cells. In liver transplant recipients, who usually receive large amounts of blood products, the effects of transfused leukocytes with regards to the rates of rejection and infection have not yet been investigated. MATERIAL/METHODS: Twenty liver transplant recipients were prospectively randomized to receive blood products (red blood cells, thrombocytes, fresh frozen plasma) through a leukocyte depletion filter, either irradiated with 40 Gy or not irradiated (10 patients, respectively). During the observation period of 90 days, the incidences of infections and rejections were analyzed. In addition, liver function tests, markers of infection (C reactive protein, lipopolysaccharide binding protein), and subpopulations of lymphocytes (total cell count, CD4/CD8 ratio, CD8CD45RO) were determined. RESULTS: Only one episode of mild rejection occurred (irradiated group). Rates of persistent graft dysfunction and severe infections were similar in both groups. The same applied to liver function tests, parameters of infection, and subpopulations of lymphocytes. CONCLUSIONS: These preliminary results suggest that irradiation of already leukocyte-depleted blood products may not be necessary and beneficial in liver transplant recipients. PMID- 17136008 TI - Serum neopterin concentrations in healthy healthcare workers compared with healthy controls and patients with pulmonary tuberculosis. AB - BACKGROUND: The aim of this study was to compare serum neopterin levels in patients with active pulmonary tuberculosis, healthy healthcare workers who had close contact with patients, and healthy volunteers. All of the healthy volunteers stated that they had not encountered possible risk factors for exposure to Mycobacterium tuberculosis infection. MATERIAL/METHODS: Thirty-nine patients, 39 healthy healthcare workers, and a control group of 39 healthy volunteers who had no infection or other diseases were included in this study. Neopterin assay was performed in accordance with the manufacturer's instructions. RESULTS: Serum neopterin levels were 18.6+/-14.2 nmol/l in patients, 9.8+/-2.9 nmol/l in healthy healthcare workers, and 3.4+/-5.2 nmol/l in healthy volunteers. Serum neopterin levels in each group were significantly different from each other (p<0.01). CONCLUSIONS: Levels of neopterin in patients and healthy healthcare workers significantly differ from the levels in healthy controls. The higher serum neopterin levels in healthy healthcare workers may be attributed to latent Mycobacterium tuberculosis infection, but it does not seem to be used in the diagnosis of latent Mycobacterium tuberculosis infection alone. However, more experiences are needed. PMID- 17136009 TI - Overexpression of aldose reductase in human cancer tissues. AB - BACKGROUND: Aldose reductase (AR) belongs to the aldo-keto reductase (AKR) super family and catalyzes the conversion of aldoses to the corresponding alcohol. Some recent studies have shown overexpression of AR and AR-like proteins in human liver cancers and some cancer cell lines such as HepG2 and HeLa cells. However, apart from hepatic cancer tissue, the status of AR expression has not been reported in other human cancer tissues. Therefore, in this preliminary report, the expression of AR in a few other commonly occurring cancer tissues was investigated. MATERIAL/METHODS: Fresh post-surgical tumor tissues of breast, ovary, cervix, and rectum were collected from subjects who were admitted for surgical therapy of tumors. Tumor area and tumor characteristics were determined by histopathological analysis. The expression and activity of AR in tumor and non tumor areas was carried out by immunohistochemical, immunoblotting, and enzyme activity studies. RESULTS: Immunoblotting results indicated overexpression of AR in breast, ovarian, cervical, and rectal cancerous tissues. Furthermore, biochemical data revealed that the specific activity of AR was higher in tumor areas than in non-tumor regions of these tissues. The overexpression of AR in tumor tissue was further validated by immunohistochemistry in the case of breast tissue. CONCLUSIONS: These preliminary results suggest overexpression and increased activity of AR in different human cancers. However, the incidence of AR overexpression and its role in drug resistance needs to be established with a large number of samples of various cancers. PMID- 17136010 TI - Sleep-disordered breathing in patients with acquired retrognathia secondary to rheumatoid arthritis. AB - BACKGROUND: Sleep-disordered breathing (SDB) is associated with a variety of conditions that cause upper-airway narrowing. It was hypothesized that upper airway narrowing can occur in patients with rheumatoid arthritis (RA) when retrognathia develops secondary to temporomandibular joint (TMJ) destruction. Therefore, the aim of this study was to detect the prevalence of SDB in patients with acquired retrognathia secondary to rheumatoid arthritis and to assess the efficacy of nasal continuous positive airway pressure (nasal CPAP) therapy in patients with SDB. MATERIAL/METHODS: Employed were a questionnaire, lateral cephalometry, and overnight polysomnography in seven women and three men (mean age +/-SD: 50+/-20 years, mean body mass index: 24.2+/-5.7 kg/m(2)) with acquired retrognathia secondary to RA. RESULTS: Three patients had severe obstructive sleep apnea (OSA) with apnea+hypopnea indices (AHI) >60/hour, three had mild obstructive sleep hypopnea (AHI >10/hour), and four had AHI <10/hour. The three patients with severe OSA all had excessive daytime sleepiness and evidence of retrognathia. In these three patients the mean AHI decreased from 72/hour to 3/hour with nasal CPAP therapy. CONCLUSIONS: SDB occurs quite frequently in non obese patients with acquired retrognathia secondary to RA. The severity of SDB is related to the degree of retrognathia and the presence of daytime sleepiness. Nasal CPAP therapy is effective and well tolerated in these patients. PMID- 17136011 TI - Memory and perceptuo-motor performance in Nigerians with chronic renal impairment. AB - BACKGROUND: With the increasing prevalence of chronic renal failure among the indigenous African population, coupled with the emphasis on improvement in the quality of life, there is a need for reports on the cognitive functioning and the effect of the disease on the cognitive performance of affected individuals. MATERIAL/METHODS: Sixty consecutive non-dialyzed Nigerian patients with clinical and biochemical evidence of chronic renal failure and sixty healthy age-, sex- and education level-matched controls (volunteers) were studied using an automated neuro-psychological test battery (FePsy) to assess their memory and perceptuo motor skills. RESULTS: The chronic renal failure patients performed worse in both verbal and visual memory than controls (p<0.0001). The patients compared favorably with the controls on concentration ability (p>0.05), but their perceptuo-motor speed was retarded (p<0.001). The levels of creatinine (p<0.01) and urea (p<0.01) and the presence of hypertension (p<0.05), asterixis (p<0.05), exertional dyspnea (p<0.05), and vomiting (p<0.05) affected their memory performance, with creatinine level being the most potent variable (p=0.0015). CONCLUSIONS: The results of this study confirm the presence of cognitive impairments in Nigerians with chronic renal impairment and buttress the importance of its aggressive and prompt management. PMID- 17136012 TI - Intraoperative epoprostenol and nitric oxide for severe pulmonary hypertension during orthotopic liver transplantation: a case report and review of the literature. AB - BACKGROUND: The presence of pulmonary hypertension in patients scheduled for liver transplantation requires a comprehensive perioperative heart evaluation and treatment with epoprostenol (prostacycline) infusion until a liver donor becomes available. We contended that intraoperative attenuation of severe pulmonary hypertension could be achieved by epoprostenol infusion combined with nitric oxide inhalation. CASE REPORT: A 49 years old man with end stage liver disease secondary to hepatitis C and ethanol abuse presented for orthotopic liver transplantation. The case was complicated by severe pulmonary hypertension. Preoperative epoprostenol, at doses ranging from 6 to 26 ng.kg(-1).min(-1), was infused during the induction of anesthesia. Although lower than before (>70 mmHg), post-induction pulmonary pressure (by Swan-Ganz catheter) was 62/30 mmHg. Prior to surgical incision nitric oxide (NO) by inhalation was commenced, increasing the concentration from 10 to 40 ppm; pulmonary artery pressure (PAP) then declined to 55/25 mmHg. Before starting reperfusion of the transplanted liver, NO concentration was increased to 80 ppm: this allowed completion of the procedure with PAP at 32/16 mmHg. Real time transesophageal echocardiography indicated improvement in right heart function due to NO. Following surgery, NO was continued for 10 hs at a concentration of 40 ppm and the patient was then extubated. Epoprostenol infusion was continued for 2 months after the patient was discharged home; last PAP was measured 32/10 mmHg. CONCLUSIONS: Severe intraoperative pulmonary hypertension during liver transplantation was successfully treated using the combination of IV epoprostenol infusion and NO inhalation in medium and high concentrations. PMID- 17136013 TI - A quadriplegic child with multiple brain abscesses: case report of neurobrucellosis. AB - BACKGROUND: This report presents a child with spinal cord lesion with concomitant multiple brain abscesses caused by brucellosis. CASE REPORT: A 12-year-old boy was admitted with quadriplegia accompanied by back pain, headache, nausea, and incontinence which appeared six months before. MRI revealed multiple brain abscesses as well as spinal intramedullary lesions. The diagnosis of brucellosis was established by positive serum Wright test (1:2560) and 2-mercaptoethanol agglutination titer (1:2560), history of exposure, and gradual improvement in response to treatment (rifampin, trimethoprim/sulfamethoxazole, gentamicin, and prednisolone). CONCLUSIONS: Neurobrucellosis may present with uncommon manifestations in children. Physicians should consider this in the differential diagnosis of a child with quadriplegia. PMID- 17136014 TI - Is the instrumentalization of humans acceptable in contemporary science? PMID- 17136015 TI - A comparative assessment of HER2 status in operable breast cancer by real-time RT PCR and by immunohistochemistry. AB - BACKGROUND: The most commonly used methods for detecting HER2 gene amplification in breast cancer are immunohistochemistry and fluorescence in-situ hybridization. The aim of this retrospective study was to assess HER2 expression by real-time RT PCR. MATERIAL/METHODS: Expression of HER2 was analyzed by real-time RT-PCR and immunohistochemistry in specimens of invasive ductal breast cancer tissue obtained from 131 women during radical mastectomy. RESULTS: There was a highly significant difference in mean relative gene expression between HER2-positive and HER2-negative patients as assessed by immunostaining (22.10+/-41.89 vs. 3.17+/ 8.76; p<0.001). With a median follow-up of 56 months, the cancer-specific survival of HER2-positive patients assessed by RT-PCR was worse in all cases and in the node-positive group. In multivariate analysis, HER2 status was an independent prognostic factor in all patients. CONCLUSIONS: Real-time RT-PCR is a valuable method for determining HER2 status, but the selection of the cut-off point of the relative gene expression differentiating between HER2-negative and positive tumors is essential. PMID- 17136016 TI - Switch to abacavir-based triple nucleoside regimens in HIV-1 infected patients never treated with suboptimal antiretroviral therapy: a review. AB - The major trials conducted on treatment simplification included large portions of patients who had previously received monotherapy or dual therapy and were likely to have relevant mutations of resistance at baseline. These studies concluded that simplification was safe (especially when this population was excluded), with some additional risk of viral failure for subjects simplified to abacavir-based regimens. On the other hand, induction-maintenance studies and other studies which involved only patients who had started HAART as the first-line showed that simplification to abacavir was as safe as continuation of the original regimen and better accepted by the patients. The largest randomized studies of simplification that allowed extrapolation of data on the population of subjects who had never received suboptimal therapy were reviewed. Four studies failed to show significant differences in efficacy between treatment arms, while two detected significant differences in favor of the continuation arms. Simplification to abacavir led to significant decreases in cholesterol and triglyceride levels and to slight improvements in quality of life. No variations in body shape were detected, although the duration was probably insufficient and most studies did not involve adequate technology (i.e. DEXA, CT). Other, smaller studies are also presented in the review, selected for their particular design or analysis, which may contribute to a better understanding of the setting in which simplification may be a feasible option. Choosing the adequate timing and the correct patient characteristics, simplification to abacavir-based regimens is safe and prevents metabolic consequences of therapy. PMID- 17136017 TI - The role of connective tissue growth factor in skeletal growth and development. AB - Connective tissue growth factor (CTGF) is a secreted, extracellular matrix associated protein that regulates diverse cellular functions in different cell types. CTGF gene belongs to a larger CCN gene family that also includes Cyr61 and NOV. It modulates many cellular functions, including proliferation, migration, adhesion, and extracellular matrix production, and it is involved in many biological and pathological processes. CTGF has special importance in skeletal development. During Meckel's cartilage development, CTGF acts as a down-stream molecule of TGFbeta to stimulate cell-cell interactions and the expression of condensation-associated genes. CTGF promotes endochondral ossification and articular cartilage regeneration. During the healing of experimental bone fracture, CTGF was expressed in periosteal cells and hypertrophic chondrocytes. It promotes the proliferation of chondrocytes and osteoblasts. CTGF is a down stream mediator for prostaglandin E2 (PGE2) in osteoblast-induced proliferation. It also regulates signaling through the Wnt pathway, in accord with its ability to bind to the Wnt co-receptor LDL receptor-related protein 6 (LRP6). Constitutive expression of CTGF was shown to inhibit both BMP-9- and Wnt3A induced osteogenic differentiation. PMID- 17136018 TI - Important aspects of Biomphalaria snail-schistosome interactions as targets for antischistosome drug. AB - The Biomphalaria species are freshwater snails which have a wide distribution and are significant both medically and economically as intermediate hosts for the schistosome parasite, a digenetic trematode causing schistosomiasis, a disease that infects 200 million people, and domestic animals throughout the tropics. The host-parasite relationship is, in principle, a powerful determinant of the biology of infection and disease. Research on snail-schistosome interactions has the potential for making an important contribution to the study of co-evolution or reciprocal adaptation. The association between Biomphalaria and Schistosoma mansoni could well be an excellent model for studies aimed at elucidating some aspects of the compatibility or resistance of this species to schistosomes. Snail hosts and schistosomes appear to have effects on each other's phenotype and genotype. The objective of this review is to clarify the nature of the relationship between schistosome parasites and their freshwater snail hosts. Aspects of snail-schistosome interactions will be traced in relation to behavioral (growth, reproduction, locomotion), immunological, and biochemical changes induced in the host's tissues by the developing intramolluscan stages of the parasite. This may help to identify biochemical or genetic targets for drug design. Manipulation of the intermediate host through these targets could break the cycle of human and snail infection by schistosomes. PMID- 17136019 TI - Free thyroxine, cognitive decline and depression in Alzheimer's disease. AB - The role of thyroid function in Alzheimer's disease (AD) has been subject to a number of studies during the last years. We investigated the possible relationship between plasma levels of the biologically active free form of thyroxin (fT4) and cognitive function in 227 outpatients with mild to moderate Alzheimer s disease (AD) in a cross-sectional study design. A significant negative correlation was found between plasma fT4-levels and Mini-Mental state examination (MMSE) score (Spearman Rho = -0.14, p=0.04). When the lowest quartile of fT4-levels (<15.1 pmol/l) was compared to the highest quartile (>19.0 pmol/l), statistically significant lower mean MMSE-scores were seen in the group with the highest fT4-levels (p<0.05, ANOVA). The mean difference between the 1st and the 4th quartile of fT4 was 2.6 MMSE-score points. No correlations were found between plasma total T4-levels, plasma total T3-levels, plasma TSH-levels and the MMSE score (p>0.05). When fT4 quartile groups were compared for depression measured in the Geriatric Depression Score (GDS 15), a slightly higher score was seen in the 1s and 2nd compared to the 3rd and 4th quartile groups without reaching statistical significance (1st quartile of fT4: GDS 5.2 +/- 3.8; 2nd: 5.3 +/- 4.0; 3rd: 4.4 +/- 3.4; 4th: 4.5 +/- 3.8) pointing to a reverse correlation of fT4 levels and depressive mood. This study leads to the conclusion that high levels of plasma fT4 might result in a worsening of cognitive impairment and a positive effect on depressive mood in AD. PMID- 17136020 TI - Missed opportunities for earlier diagnosis of HIV infection--South Carolina, 1997 2005. AB - In September 2006, CDC published revised recommendations for human immunodeficiency virus (HIV) testing in health-care settings to 1) increase early detection of HIV infection by expanding HIV screening of patients and 2) improve access to HIV care and prevention services (e.g., by conducting screening in locations such as emergency departments and urgent-care facilities, where persons who do not otherwise access HIV testing seek health-care services). HIV screening is now recommended for patients aged 13-64 years in all health-care settings after patients are notified that testing will be performed unless they decline (opt-out screening). This represents a substantial change from earlier recommendations to 1) offer HIV testing routinely to all patients only in health care settings with high HIV prevalence and 2) conduct targeted screening on the basis of risk behaviors for patients in low-prevalence settings. This report examines HIV and acquired immunodeficiency syndrome (AIDS) case reporting in South Carolina before the 2006 recommendations were published. During 2001-2005, a total of 4,315 cases of HIV infection were reported in South Carolina. Of these, 41% were in persons (referred to as late testers) in whom AIDS was diagnosed within 1 year of their initial HIV diagnosis. Of these late testers, 73% made a total of 7,988 visits to a South Carolina health-care facility during 1997-2005 before their first reported positive HIV test. The diagnoses reported for 79% of these visits were not likely to prompt HIV testing under a risk-based testing strategy. These findings suggest that routine, opt-out HIV screening of all patients in health-care settings, rather than risk-based HIV testing, might result in substantially earlier HIV diagnoses in South Carolina. PMID- 17136021 TI - Injuries from motor-vehicle collisions with moose--Maine, 2000-2004. AB - Moose are among the largest mammals in North America. Standing up to 7.5 feet at the shoulder and weighing up to 1,600 lbs, they are the largest members of the deer family. Maine's moose population (approximately 29,000) is the biggest in the United States outside of Alaska. During a collision with a motor vehicle, a moose usually is struck in the legs, causing its body to roll onto the hood of the vehicle, often collapsing the windshield and roof. As a result, motor-vehicle collisions involving moose are capable of causing substantial injury to vehicle occupants. To assess motor-vehicle collisions with moose in Maine and evaluate risk factors for injuries from these types of collisions, the Maine Department of Health and Human Services studied collision reports from 2000--2004. The results of that study indicated that collision rates varied by county but had clear patterns by season and time of day. Variables associated with risk for injury were posted speed limit, type of vehicle, and sex and age of the driver. Measures to reduce collisions with moose should focus on improving driver education programs and developing better engineering controls (e.g., removing roadside vegetation to improve visibility for drivers). In addition, herd management (i.e., decreasing moose population size through hunting) is currently being used in areas of Maine with high numbers of collisions, although studies are needed to assess its effectiveness. PMID- 17136022 TI - Racial/ethnic differences among youths in cigarette smoking and susceptibility to start smoking--United States, 2002-2004. AB - Limited information on cigarette smoking in racial/ethnic subpopulations hinders development and implementation of targeted interventions for smoking prevention and cessation. Because of small sample sizes or inadequate study formats, cigarette smoking among youths has been studied mostly in major racial/ethnic populations (e.g., Asian or Hispanic) instead of subsets of these populations (e.g., Vietnamese or Cuban). Data on major population categories might mask differences in tobacco-use prevalence among subpopulations. To assess the prevalence of cigarette smoking among youths aged 12-17 years in six major racial/ethnic populations and nine Asian or Hispanic subpopulations in the United States, the Substance Abuse and Mental Health Services Administration and CDC analyzed self-reported data collected during 2002-2004 from the National Survey on Drug Use and Health (NSDUH). This report summarizes the results of that analysis, which indicated that the estimated prevalence of cigarette smoking in this age group ranged from 23.1% for American Indians/Alaska Natives (AI/ANs) to 2.2% for Vietnamese. Implementing tobacco-control programs that include culturally appropriate interventions might help reduce cigarette smoking in racial/ethnic subpopulations. PMID- 17136023 TI - Brief report: respiratory syncytial virus activity--United States, 2005-2006. AB - Respiratory syncytial virus (RSV) is a major cause of lower respiratory tract infections (LRTIs) (e.g., bronchiolitis and pneumonia) among young children in the United States. RSV also causes severe respiratory disease and a substantial number of deaths among older adults and persons with compromised respiratory, cardiac, or immune systems. RSV is transmitted person to person through close contact or inhalation of large droplets from a sneeze or cough; infection also can occur through contact with fomites (i.e., contaminated surfaces or objects). In temperate climates, peak RSV activity typically occurs during the winter. This report presents preliminary data on RSV activity reported to the National Respiratory and Enteric Virus Surveillance System (NREVSS) for the weeks ending July 8-November 18, 2006, indicating the onset of the 2006-2007 RSV season, and summarizes RSV trends during July 2005-June 2006. Health-care providers should consider RSV in the differential diagnosis for persons of all ages with LRTIs and implement appropriate isolation precautions to prevent nosocomial transmission from RSV-infected patients. Immune prophylaxis should be considered for certain infants and young children at high risk for complications from RSV infection (e.g., certain premature infants or infants and children with chronic lung and heart disease). PMID- 17136024 TI - General recommendations on immunization: recommendations of the Advisory Committee on Immunization Practices (ACIP). AB - This report is a revision of General Recommendations on Immunization and updates the 2002 statement by the Advisory Committee on Immunization Practices (ACIP) (CDC. General recommendations on immunization: recommendations of the Advisory Committee on Immunization Practices and the American Academy of Family Physicians. MMWR 2002;51[No. RR-2]). This report is intended to serve as a general reference on vaccines and immunization. The principal changes include 1) expansion of the discussion of vaccination spacing and timing; 2) an increased emphasis on the importance of injection technique/age/body mass in determining appropriate needle length; 3) expansion of the discussion of storage and handling of vaccines, with a table defining the appropriate storage temperature range for inactivated and live vaccines; 4) expansion of the discussion of altered immunocompetence, including new recommendations about use of live-attenuated vaccines with therapeutic monoclonal antibodies; and 5) minor changes to the recommendations about vaccination during pregnancy and vaccination of internationally adopted children, in accordance with new ACIP vaccine-specific recommendations for use of inactivated influenza vaccine and hepatitis B vaccine. The most recent ACIP recommendations for each specific vaccine should be consulted for comprehensive discussion. This report, ACIP recommendations for each vaccine, and other information about vaccination can be accessed at CDC's National Center for Immunization and Respiratory Diseases (proposed) (formerly known as the National Immunization Program) website at http//:www.cdc.gov/nip. PMID- 17136027 TI - Vascular function in patients with end-stage renal disease and/or coronary artery disease: a cardiac magnetic resonance imaging study. AB - Decreased arterial compliance in end-stage renal disease (ESRD) is associated with increased cardiovascular risk. Our aim was to examine aortic compliance in patients with ESRD using cardiac magnetic resonance imaging (MRI) and to compare these with patients with advanced atherosclerotic disease who are known to be at high cardiovascular risk. We examined a total of 83 subjects matched for age: 24 had ESRD and were on dialysis therapy for 3+/-6 years, 24 had severe coronary artery disease (CAD), 11 had both ESRD and CAD (4+/-5 years on dialysis therapy), and 24 healthy subjects with no evidence of CAD. Vascular and cardiac function was assessed using cardiac MRI. Aortic compliance was significantly reduced in patients with CAD compared to control subjects (11.3+/-6.3 ml x 10(-3)/mm Hg vs 15.6+/-6.0 ml x 10(-3)/mm Hg, P=0.009). Patients with ESRD also exhibited significantly reduced aortic compliance compared to healthy controls (12.4+/-5.8 ml x 10(-3)/mm Hg vs 15.6+/-6.0 ml 10(-3)/mm Hg, P=0.012), whereas there was no significant difference in aortic compliance between patients with CAD and ESRD. Even in the absence of symptomatic CAD, patients with ESRD have significantly reduced aortic compliance compared to normal subjects. Patients with ESRD have equivalent aortic compliance to patients with advanced CAD. These findings suggest that a significantly reduced aortic compliance is one of many mechanisms promoting premature cardiovascular events in patients with ESRD compared to age matched controls from the general population. PMID- 17136028 TI - Non-invasive monitoring of kidney allograft rejection through IDO metabolism evaluation. AB - The immunomodulatory enzyme indoleamine 2,3-dioxygenase (IDO) is activated by interferon-gamma (IFN-gamma) and via tryptophan depletion, suppresses adaptive T cell-mediated immunity in inflammation, host immune defense, and maternal tolerance. Its role in solid organ transplantation is still unclear. Therefore, we investigated the usefulness of IDO-mediated tryptophan catabolism in the evaluation of kidney allograft rejection. Blood, urine, and tissue samples were collected from 34 renal transplant patients without rejection and from nine patients with biopsy-confirmed episodes of acute rejection (n=12). Concentrations of kynurenine and tryptophan in serum and urine were analyzed by high-pressure liquid chromatography. Kynurenine to tryptophan ratio (kyn/trp) was calculated to estimate IDO activity. Immunostaining for IDO was performed on renal biopsies. Neopterin was assessed using radioimmunoassay. Kyn/trp and neopterin were detectable at low levels in serum of healthy volunteers and were increased in non rejecting allograft recipients. Serum levels of kyn/trp were higher in recipients with rejection compared to non-rejectors as early as by day 1 post-surgery. Rejection episodes occurring within 13+/-5.9 days after transplantation were accompanied by elevated kyn/trp in serum (114+/-44.5 micromol/mmol, P=0.001) and urine (126+/-65.9 micromol/mmol, P=0.02) compared to levels during stable graft function. Kyn/trp correlated significantly with neopterin suggesting an IFN-gamma induced increase in IDO activity. Immunostaining showed upregulation of IDO in rejection biopsies, localized in tubular-epithelial cells. Non-rejected grafts displayed no IDO expression. Acute rejection is associated with simultaneously increased serum and urinary kyn/trp in patients after kidney transplantation. Thus, IDO activity might offer a novel non-invasive means of immunomonitoring of renal allografts. PMID- 17136029 TI - Spontaneous leukocyte activation and oxygen-free radical generation in end-stage renal disease. AB - Oxidative stress and inflammation are common features and major mediators of atherosclerosis in end-stage renal disease (ESRD). Available evidence for oxidative stress in ESRD is indirect and based on accumulation of byproducts of interactions of reactive oxygen species (ROS) with various molecules. Inflammation is a major cause of oxidative stress. To explore the direct link between oxidative stress and inflammation in ESRD, we studied leukocyte integrin expression and ROS production in 18 ESRD patients and 18 controls. ESRD patients showed elevated plasma malondialdehyde (MDA) and increased superoxide and hydrogen peroxide (H(2)O(2)) production by granulocytes and monocytes before dialysis. Hemodialysis resulted in a further rise in plasma MDA and H(2)O(2) production by granulocytes and monocytes. Surface expression of Mac-1 (CD11b and CD18) on granulocytes and monocytes was significantly increased (denoting cell activation) in ESRD patients. Granularity of granulocytes was significantly reduced before dialysis and declined further after dialysis. The magnitude of ROS production by granulocytes and monocytes was directly related with CD11b expression as well as plasma ferritin and parathyroid hormone levels and was inversely related to protein catabolic rate. Thus, this study provides direct evidence of spontaneous leukocyte activation and increased ROS generation (hence the link between oxidative stress and inflammation) in ESRD patients. PMID- 17136030 TI - Risk factors for chronic kidney disease in a community-based population: a 10 year follow-up study. AB - The purpose of this study was to explore risk factors affecting the incidence of chronic kidney disease (CKD) in general population. We conducted a 10-year follow up study with 123 764 (male: 41 012, female: 82 752) adults aged 40 years and over who received community-based annual examinations. The primary outcome for the analysis was the development of CKD during the follow-up period. Predictors for the development of CKD were obtained by the significant hazard ratios (HR) in Cox regression model by sex. During the follow-up period, 4307 subjects (male: 2048, female: 2259) developed CKD stage I or II, and 19 411 subjects (male: 4257, female: 15 154) developed CKD stage III or higher. The baseline-adjusted predictor of developing CKD included age, glomerular filtration rate, hematuria, hypertension, diabetes, serum lipids, obesity, smoking status, and consumption of alcohol. Treated diabetes in male subjects, and treated hypertension, systolic blood pressure >160 mm Hg and/or diastolic blood pressure >100 mm Hg, diabetes, and treated diabetes in female subjects were associated with more than a doubling of the HR. For the development of CKD stage III or higher, proteinuria of >or= + +, and proteinuria and hematuria were associated with more than a doubling of the HR in male subjects. The prevalence of newly developed CKD over 10 years was 23 718 subjects (19.2%) in adults. This study suggested that not only hypertension and diabetes but also several metabolic abnormalities were independent risk factors for developing CKD. PMID- 17136031 TI - Comparison of hemodialysis and peritoneal dialysis survival in The Netherlands. AB - Considerable geographic variation exists in the relative use of hemodialysis (HD) vs peritoneal dialysis (PD). Studies comparing survival between these modalities have yielded conflicting results. Our aim was to compare the survival of Dutch HD and PD patients. We developed Cox regression models using 16 643 patients from the Dutch End-Stage Renal Disease Registry (RENINE) adjusting for age, gender, primary renal disease, center of dialysis, year of start of renal replacement therapy, and included several interaction terms. We assumed definite treatment assignment at day 91 and performed an intention-to-treat analysis, censoring for transplantation. To account for time dependency, we stratified the analysis into three time periods, >3-6, >6-15, and >15 months. For the first period, the mortality hazard ratio (HR) of PD compared with HD patients was 0.26 (95% confidence interval (CI) 0.17-0.41) for 40-year-old non-diabetics, which increased with age and presence of diabetes to 0.95 (95% CI 0.64-1.39) for 70 year-old patients with diabetes as primary renal disease. The HRs of the second period were generally higher. After 15 months, the HR was 0.86 (95% CI 0.74-1.00) for 40-year-old non-diabetics and 1.42 (95% CI 1.23-1.65) for 70-year-old patients with diabetes as primary renal disease. We conclude that the survival advantage for Dutch PD compared with HD patients decreases over time, with age and in the presence of diabetes as primary disease. PMID- 17136032 TI - Emphysematous pyelonephritis. PMID- 17136033 TI - Associations of serum minerals with body mass index in adult women. AB - OBJECTIVE: This cross-sectional study evaluated the association between serum minerals and body mass index in adult women. METHODS: One hundred and eighteen adult women were recruited by written advertisement from outpatient clinics or a health promotion center at a university hospital. Serum calcium, magnesium, copper and zinc were measured by an automatic analytical instrument and body mass index was calculated from height and weight. RESULTS: Serum magnesium was inversely associated with body mass index (beta=-0.283, P=0.001) whereas serum copper had a positive association with body mass index (beta=0.197, P=0.025) after adjusting for age, physical activity, energy intake, dietary fat, alcohol consumption, supplements and menopause status. No associations were found with serum calcium and zinc. CONCLUSION: Serum magnesium and copper may be involved in the regulation of body size in adult women. PMID- 17136034 TI - Effects of the glycemic index of breakfast on metabolic responses to brisk walking in females. AB - OBJECTIVE: The aim of this study was to examine the metabolic responses during 1 h of brisk walking, 3 h after ingesting high glycemic index (HGI) and moderate glycemic index (MGI) breakfasts. DESIGN AND SUBJECTS: Six females completed three treadmill walking trials (approximately 50% VO2 max), separated by at least 1 month. Three hours before walking, they ingested either water or a HGI or MGI breakfast. The MGI breakfast consisted of a mixture of an HGI breakfast cereal and low GI carbohydrate (CHO) foods. The GI values of the HGI and MGI meals were 77 and 51, respectively. SETTING: The study took place in the School of Sport and Exercise Sciences at Loughborough University, Loughborough, UK. RESULTS: In the HGI and MGI trials, plasma glucose and serum insulin concentrations peaked 15 min into the postprandial period. At the onset of exercise, plasma insulin concentrations were twofold higher in the HGI (31.5+/-7.7 microl U l(-1)) than in the MGI trial (15.2+/-1.9 microl U l(-1)) (P<0.05). However, there were no differences in substrate utilization between the two CHO trials. CONCLUSION: These results suggest that although the addition of LGI CHO foods to an HGI breakfast cereal reduces the overall GI of the meal, the metabolic response to exercise is similar to that following a breakfast comprised entirely of HGI foods. PMID- 17136035 TI - Nutritional implications of patient-provider interactions in hospital settings: evidence from a within-subject assessment of mealtime exchanges and food intake in elderly patients. AB - OBJECTIVE: To examine the nutritional implications of the interactions taking place between patients and care providers during mealtimes in hospital settings. Specifically, we tested research propositions that the amount and nature of interpersonal behaviours exchanged between patients and providers impact patients' food intake. These propositions were derived from prior evidence of social influences on eating behaviour and a well-established framework that identifies two fundamental modalities of human interaction: striving for mastery and power (agency) and efforts to promote union with others (communion). DESIGN: In a within-subject naturalistic study, participants were observed on multiple meals (n=1477, 46.2 meals/participant on average), during which participants' and providers' agency- and communion-related behaviours and patients' protein and energy intake were recorded. Meal-level frequency and complementarity of patients' and providers' behaviours were computed to test research propositions. SETTING: Dining room of a geriatric rehabilitation unit. SUBJECTS: Thirty-two elderly patients (21 females, mean age:78.8, 95% CI: 76.4, 81.1). RESULTS: Meal level frequency of patient-provider exchanges (P=0.016) and patients' agency related behaviours (P=0.029), as well as mutual reciprocation of patients' and providers' communion-related behaviours (P=0.015) on a given meal were positively linked to protein intake. Higher energy intake was found during meals where patients expressed more agency-related behaviours (P=0.029). CONCLUSION: Results present evidence that the amount and nature of patient-provider interpersonal exchanges on a given meal influence the nutritional quality of food intake in hospitalized elderly. They provide insights into how to improve the design and delivery of routine care to this malnutrition-prone population. SPONSORSHIP: This study was supported by the Canadian Institutes of Health Research (Operating grant to Laurette Dube, Doctoral Fellowship to Catherine Paquet) the Fonds de la Recherche en sante du Quebec and by the Danone Institute (Doctoral fellowship to Danielle St-Arnaud McKenzie). PMID- 17136036 TI - Overweight with concurrent stunting in very young children from rural Mexico: prevalence and associated factors. AB - OBJECTIVE: To document the prevalence of overweight or obesity concurrent with stunting in rural low-income Mexican children and to identify demographic and socio-economic characteristics that could help identify families at risk of having an overweight/obese and stunted young child in this population. DESIGN: Cross-sectional analysis of the nutritional status of very young children, using primary data from a rural community-based survey conducted in 2003. Overweight, obesity and stunting were documented along with several maternal, household and community characteristics. SETTING: Impoverished areas of rural Mexico. SUBJECTS: Pre-school children (n=7555), aged 24-72 months. RESULTS: The combined prevalence of overweight and obesity was equal to or greater than 20% in all children, as was the prevalence of stunting. The prevalence of concurrent overweight or obesity and stunting was approximately 5% in non-indigenous children, and over 10% in indigenous children 24-60 months. A multinomial logistic analysis revealed that the factors associated with coexisting stunting and overweight/obesity were lower socio-economic status (SES), lower maternal age, education, intelligence (vocabulary) and perceived social status, shorter maternal height, and larger household size. Among only stunted children, the risk of also being overweight or obese was associated with younger maternal age (relative risk ratios (RRR): 0.98, P=0.05), lower maternal perceived social status (RRR: 0.95, P<0.01) and maternal obesity (RRR: 2.93, P<0.0001) or overweight (RRR: 1.50, P=0.002). CONCLUSIONS: These analyses highlight that concurrent overweight or obesity and stunting is an important public health issue in low-income areas of rural Mexico beginning in early childhood. Even within this impoverished population, children living in households with low relative SES are the most vulnerable. SPONSORSHIP: Financial support for this research was provided by the National Institutes of Child Health and Human Development, the Fogarty International Center at NIH, the John D and Catherine T MacArthur Foundation 'Research Network on Socioeconomic Status and Health' and the Mexican Government. PMID- 17136037 TI - Low-carbohydrate-high-protein diet and long-term survival in a general population cohort. AB - OBJECTIVE: We have evaluated the effects on mortality of habitual low carbohydrate-high-protein diets that are thought to contribute to weight control. DESIGN: Cohort investigation. SETTING: Adult Greek population. SUBJECTS METHODS: Follow-up was performed from 1993 to 2003 in the context of the Greek component of the European Prospective Investigation into Cancer and nutrition. Participants were 22 944 healthy adults, whose diet was assessed through a validated questionnaire. Participants were distributed by increasing deciles according to protein intake or carbohydrate intake, as well as by an additive score generated by increasing decile intake of protein and decreasing decile intake of carbohydrates. Proportional hazards regression was used to assess the relation between high protein, high carbohydrate and the low carbohydrate-high protein score on the one hand and mortality on the other. RESULTS: During 113 230 persons years of follow-up, there were 455 deaths. In models with energy adjustment, higher intake of carbohydrates was associated with significant reduction of total mortality, whereas higher intake of protein was associated with nonsignificant increase of total mortality (per decile, mortality ratios 0.94 with 95% CI 0.89 0.99, and 1.02 with 95% CI 0.98 -1.07 respectively). Even more predictive of higher mortality were high values of the additive low carbohydrate-high protein score (per 5 units, mortality ratio 1.22 with 95% CI 1.09 -to 1.36). Positive associations of this score were noted with respect to both cardiovascular and cancer mortality. CONCLUSION: Prolonged consumption of diets low in carbohydrates and high in protein is associated with an increase in total mortality. PMID- 17136038 TI - Influence of methods used in body composition analysis on the prediction of resting energy expenditure. AB - OBJECTIVE: There are considerable differences in published prediction algorithms for resting energy expenditure (REE) based on fat-free mass (FFM). The aim of the study was to investigate the influence of the methodology of body composition analysis on the prediction of REE from FFM. DESIGN: In a cross-sectional design measurements of REE and body composition were performed. SUBJECTS: The study population consisted of 50 men (age 37.1+/-15.1 years, body mass index (BMI) 25.9+/-4.1 kg/m2) and 54 women (age 35.3+/-15.4 years, BMI 25.5+/-4.4 kg/m2). INTERVENTIONS: REE was measured by indirect calorimetry and predicted by either FFM or body weight. Measurement of FFM was performed by methods based on a 2 compartment (2C)-model: skinfold (SF)-measurement, bioelectrical impedance analysis (BIA), Dual X-ray absorptiometry (DXA), air displacement plethysmography (ADP) and deuterium oxide dilution (D2O). A 4-compartment (4C)-model was used as a reference. RESULTS: When compared with the 4C-model, REE prediction from FFM obtained from the 2C methods were not significantly different. Intercepts of the regression equations of REE prediction by FFM differed from 1231 (FFM(ADP)) to 1645 kJ/24 h (FFM(SF)) and the slopes ranged between 100.3 kJ (FFM(SF)) and 108.1 kJ/FFM (kg) (FFM(ADP)). In a normal range of FFM, REE predicted from FFM by different methods showed only small differences. The variance in REE explained by FFM varied from 69% (FFM(BIA)) to 75% (FFM(DXA)) and was only 46% for body weight. CONCLUSION: Differences in slopes and intercepts of the regression lines between REE and FFM depended on the methods used for body composition analysis. However, the differences in prediction of REE are small and do not explain the large differences in the results obtained from published FFM-based REE prediction equations and therefore imply a population- and/or investigator specificity of algorithms for REE prediction. PMID- 17136039 TI - Blood pressure among overweight adolescents from urban school children in Pune, India. AB - OBJECTIVES: To examine the magnitude of overweight and its association with blood pressure (BP) among adolescents. DESIGN: Cross-sectional study with all children in age range 9-16 years (n=1146 boys and 1077 girls) from two schools catering to urban affluent high socio-economic class (HSE), for anthropometric measurements by trained investigators and BP measurement by a pediatrician using sphygmomanometer. RESULTS: The prevalence of overweight based on conventional body mass index (BMI) cutoff was 27.5% for boys and 20.9% for girls but varied for different indicators. Prevalence of high systolic blood pressure (HSBP) was 12.0% in boys and 9.7% in girls and increased with increasing levels of BMI, weight, triceps skin fold thickness (TSFT) and percent body fat. Mean level of SBP among overweight children was significantly (P<0.001) higher by about 12 mm Hg, whereas that for diastolic blood pressure was higher by 8 mm Hg (P<0.001) as compared to their non-overweight (age, sex-matched) counterparts. This was true in both sexes and for all indicators used for assessing overweight. Prevalence of HSBP increased suddenly beyond BMI value of 20 kg/m2 in boys and 21.5 kg/m2 in girls, beyond TSFT value of 12 mm for boys and 14 mm for girls whereas such cutoffs for body fat were above 25% in both sexes. These cutoffs appear much lower than the conventional ones and therefore indicate the need for validation of conventional cutoffs in different populations. CONCLUSIONS: Our findings highlight that BP measurement needs to be a routine part of physical examination in school children, and the use of cutoffs anchored to metabolic risks may be essential for assessment of obesity. PMID- 17136040 TI - Efficacy of alphas1-casein hydrolysate on stress-related symptoms in women. AB - OBJECTIVE: To examine the effects of alpha (s1)-casein hydrolysate on females with stress-related symptoms. DESIGN: Double-blind, randomized, crossover, placebo-controlled trial. SETTING: The alpha (s1)-casein hydrolysate was manufactured by INGREDIA (Arras, France) and the placebo was manufactured by DIETAROMA (Bourg, France). Study was designed and performed at PROCLAIM (Rennes, France), and the statistical analyses were performed by D Desor (Nancy, France). SUBJECTS: A total of 63 female volunteers suffering from at least one disorder that may be related to stress such as anxiety, sleep problems and general fatigue. INTERVENTIONS: A total of 63 volunteers participated in a double-blind, randomized, crossover, placebo-controlled study. Subjects were randomly allocated to receive either tablets containing alpha (s1)-casein hydrolysate or placebo at the dose of 150 mg/day for 30 days. After a 3 weeks washout period, they were crossed over for a new 30-day period of tablets intake. The outcome measure was a questionnaire including 44 items of symptoms that may be related stress in which the severity of each sign was evaluated using a 10-degree scale. These measures were studied repeatedly at the day of 0, 15 and 30 after the start of each interventional period. RESULTS: The 30-day treatment by alpha (s1)-casein hydrolysate in females with stress-related symptoms reduced their symptoms, particularly in digestion (P<0.01), cardiovascular (P<0.05), intellectual (P<0.01), emotional (P<0.05) and social problems (P<0.05). CONCLUSION: This study showed that a 30-day ingestion of alpha (s1)-casein hydrolysate decreased the stress-related symptoms in females suggesting that this product may be used as an effective functional ingredient alleviating such symptoms. SPONSORSHIP: This study was partially supported by the INGREDIA of France and Neurobiology Research Program from the Korea Ministry of Science and Technology (2004-01757) of Korea. PMID- 17136041 TI - Higher maternal dietary protein intake in late pregnancy is associated with a lower infant ponderal index at birth. AB - AIM: A high ponderal index at birth has been associated with later obesity and it has been suggested that intervention to prevent obesity and its sequela should consider the antenatal period. In this context, we investigated the association between maternal nutrition and birth anthropometry. DESIGN: We analyzed data on 1040 mother-infant pairs collected during the Tasmanian Infant Health Survey (TIHS), Tasmania, 1988-1989. Maternal dietary intake during pregnancy was measured by food frequency questionnaire (FFQ) applied soon after birth. Outcomes of interest were birth weight, birth length, head circumference, ponderal index, head circumference -to-ponderal index ratio, placenta-to-birth weight ratio and head circumference-to-birth length index. RESULTS: In multiple regression model, an increase of 10 g of absolute protein intake/day was associated with a reduction in birth weight of 17.8 g (95% CI: -32.7, -3.0; P=0.02). Protein intake was also associated negatively with ponderal index (beta=-0.01; 95% CI: -0.02, 0.00; P=0.01). A 1 % increase in carbohydrate intake resulted in a 1% decline in placental weight relative to birth weight. Higher protein intake in the third trimester was associated with a reduced ponderal index among large birth weight infants but not low birth weight infants. CONCLUSIONS: This raises the possibility that any effect of high protein in altering infant anthropometry at birth may involve changes in body composition and future work to examine how a high-protein diet influences body composition at birth is warranted. PMID- 17136042 TI - Effect on hematologic risk factors for coronary heart disease of a cholesterol reducing diet. AB - BACKGROUND: A dietary portfolio of cholesterol-lowering ingredients has proved effective in reducing serum cholesterol. However, it is not known whether this dietary combination will also affect hematologic risk factors for coronary heart disease (CHD). Reductions in hematocrit and polymorphonuclear leukocytes have been reported to improve cardiovascular risk. We, therefore, report changes in hematological indices, which have been linked to cardiovascular health, in a 1 year assessment of subjects taking an effective dietary combination (portfolio) of cholesterol-lowering foods. METHODS: For 12 months, 66 hyperlipidemic subjects were prescribed diets high in plant sterols (1.0 g/1000 kcal), soy protein (22.5 g/1000 kcal), viscous fibers (10 g/1000 kcal) and almonds (23 g/1000 kcal). Fifty five subjects completed the study. RESULTS: Over the 1 year, data on completers indicated small but significant reductions in hemoglobin (-1.5+/-0.6 g/l, P=0.013), hematocrit (-0.007+/-0.002 l/l, P<0.001), red cell number (-0.07+/-0.02 10(9)/l, P<0.001) and neutrophils (-0.34+/-0.13 10(9)/l, P=0.014). Mean platelet volume was also increased (0.16+/-0.07 fl, P=0.033). The increase in red cell osmotic fragility (0.05+/-0.03 g/l, P=0.107) did not reach significance. CONCLUSIONS: These small changes in hematological indices after a cholesterol lowering diet are in the direction, which would be predicted to reduce CHD risk. Further research is needed to clarify whether the changes observed will contribute directly or indirectly to cardiovascular benefits beyond those expected from reductions previously seen in serum lipids and blood pressure. PMID- 17136043 TI - Vitamin D fortification of milk products does not resolve hypovitaminosis D in young Finnish men. AB - OBJECTIVE: To study if vitamin D fortification of milk products started in February 2003 has improved vitamin D status of young Finnish men, which has been poor before. DESIGN: A longitudinal study of one cohort. SETTING: Helsinki University Central Hospital. SUBJECTS: Sixty-five healthy men, studied for the first time in January 2001, were re-examined in January 2004. They were aged 18 21 years in 2001. METHODS: Blood was sampled for determination of serum 25 hydroxyvitamin D (25-OHD) and intact parathyroid hormone (iPTH). 25-OHD was measured by both radioimmunoassay (RIA) and high-pressure liquid chromatography (HPLC). Consumption of milk, sour milk and fish and use of vitamin D supplements were assessed using a questionnaire. RESULTS: In January 2004, vitamin D fortification had raised serum 25-OHD level, with the mean of individual percent changes being 20.4% measured with RIA (P=0.0015). The correlation between the RIA and HPLC methods was high (r=0.85). Nineteen men (29.2%) had vitamin D deficiency (25-OHDG (I22M) in the 5 methyltetrahydrofolate-homocysteine methyltransferase reductase (MTRR) gene showed any effect on disease risk. The carriers of the G-allele were associated with a marginal decreased risk of ALL (gender-adjusted global P=0.03; multiple testing corrected P=0.25). Analysis of four polymorphisms in the MTRR gene showed statistically significant differences in haplotype distribution between cases and controls (global P<0.0001). The haplotypes GCAC (odds ratio (OR) 0.5, 95% confidence interval (CI) 0.4-0.6) and ATAC (OR 0.5, 95% CI 0.3-0.6) were associated with a reduced risk and the haplotypes ACAC (OR 2.3, 95% CI 1.8-2.9) and GTAC (OR 1.8, 95% CI 1.4-2.3) with an increased risk. The genotype combination analyses indicated that the best model stratifies cases and controls based on the 66A>G and the 524C>T polymorphisms in the MTRR gene (global P=0.03). Our results suggest that, besides a weak association of childhood ALL with the 66A>G polymorphism, haplotypes within the MTRR gene may, in part, account for population-based differences in risk. PMID- 17136116 TI - Antiproliferative activity of RAD001 (everolimus) as a single agent and combined with other agents in mantle cell lymphoma. AB - Mantle cell lymphoma (MCL) is an aggressive form of B-cell non-Hodgkin's lymphoma, with a mean survival of only 3-5 years and suboptimal therapeutic options. MCL is characterized by a balanced translocation t(11;14)(q13;q32), resulting in overexpression of cyclin D1, a G(1) cyclin regulated by the PI3K/Akt/mammalian target of rapamycin (mTOR) signaling pathway. As improved therapy for MCL is required and the mTOR pathway may be involved in its pathophysiology, the antiproliferative effects of RAD001 (everolimus), an mTOR inhibitor, against three MCL cell lines were investigated. As a single agent, RAD001 inhibited proliferation in MCL cell lines (Jeko1, SP49 and NCEB1) approximately 40-65% compared to diluent control cells. This was associated with G(1) cell-cycle arrest and reduced phosphorylation of the mTOR downstream target, 4E-BP1. Furthermore, combination drug studies revealed predominantly synergistic cytotoxicity with RAD001 and several secondary agents, including doxorubicin, vincristine or rituximab (components of the standard MCL regimen), as well as paclitaxel, vorinostat and bortezomib. These data indicate that single agent RAD001 is effective in inhibiting growth of MCL cells in vitro and combination studies with secondary agents further demonstrate synergistic cytotoxicity. Thus, these findings support future clinical studies of RAD001 in the treatment of MCL. PMID- 17136117 TI - Morphological and molecular characterization of novel population of CXCR4+ SSEA 4+ Oct-4+ very small embryonic-like cells purified from human cord blood: preliminary report. AB - Recently, we purified from adult murine bone marrow (BM) a population of CXCR4(+), Oct-4(+) SSEA-1(+), Sca-1(+) lin(-) CD45(-) very small embryonic-like (VSEL) stem cells and hypothesized that similar cells could be also present in human cord blood (CB). Here, we report that by employing a novel two-step isolation procedure -- removal of erythrocytes by hypotonic lysis combined with multiparameter sorting -- we could isolate from CB a population of human cells that are similar to murine BM-derived VSELs, described previously by us. These CB isolated VSELs (CB-VSEL) are very small (3-5 micro m) and highly enriched in a population of CXCR4(+)AC133(+)CD34(+)lin(-) CD45(-) CB mononuclear cells, possess large nuclei containing unorganized euchromatin and express nuclear embryonic transcription factors Oct-4 and Nanog and surface embryonic antigen SSEA-4. Further studies are needed to see if human CB-isolated VSELs similar to their murine BM-derived counterparts are endowed with pluripotent stem cell properties. PMID- 17136122 TI - Genome-wide linkage analysis of 160 North American families with celiac disease. AB - Celiac disease (CD) is a common autoimmune disease caused by exposure to the protein gliadin in wheat, and related prolamins in barley and rye. The prevalence of the disease in the US is 1:133. The aim of this study was to identify non human leukocyte antigen (HLA) loci that predispose to CD. A genome-wide search of 405 microsatellite markers was performed on DNA samples from 160 families with a minimum of two cases of CD. Multipoint, parametric and non-parametric linkage (NPL) analyses were performed. Locations on chromosomes 1q, 3q, 6p, 6q, 7q, 9q and 10q showed linkage statistics (NPL scores or heterogeneity logarithm of the odds (HLOD) scores) of approximately 2.0 or larger. The greatest evidence for linkage outside of chromosome 6 was on 7q and 9q. An NPL score of 2.60 occurred at position 151.0 on 7q and a HLOD score of 2.47 occurred at position 144.8 on 9q under a recessive model. As expected, there was highly significant linkage to the HLA region on 6p, with NPL and HLOD scores exceeding 5.50. In conclusion, this genome-wide linkage analysis represents one of the largest such studies of CD. The most promising region is a putative locus on 7q, a region reported independently in previous genome-wide searches. PMID- 17136123 TI - No evidence of association between genetic variants of the PDCD1 ligands and SLE. AB - PDCD1, an immunoreceptor involved in peripheral tolerance has previously been shown to be genetically associated with systemic lupus erythematosus (SLE). PDCD1 has two ligands whose genes are located in close proximity on chromosome 9p24. Our attention was drawn to these ligands after finding suggestive linkage to a marker (gata62f03, Z=2.27) located close to their genes in a genome scan of Icelandic families multiplex for SLE. Here, we analyse Swedish trios (N=149) for 23 single nucleotide polymorphisms (SNPs) within the genes of the PDCD1 ligands. Initially, indication of association to eight SNPs was observed, and these SNPs were therefore also analysed in Mexican trios (N=90), as well as independent sets of patients and controls from Sweden (152 patients, 448 controls) and Argentina (288 patients, 288 controls). We do not find support for genetic association to SLE. This is the first genetic study of SLE and the PDCD1 ligands and the lack of association in several cohorts implies that these genes are not major risk factors for SLE. PMID- 17136124 TI - IFNG and IFNGR1 gene polymorphisms and susceptibility to post-kala-azar dermal leishmaniasis in Sudan. AB - Post-kala-azar dermal leishmanaisis (PKDL) in Sudan is associated with elevated interferon-gamma (IFN-gamma). To study interferon-gamma pathways in PKDL, we genotyped 80 trios from the Masalit ethnic group for polymorphisms at -470 ins/delTT, -270T/C, -56T/C and +95T/C in IFNGR1 and at -179G/A and +874T/A in IFNG. No associations occurred at IFNG. Global association with haplotypes comprising all four markers at IFNGR1 (chi(2)(10df)=21.97, P=0.015) was observed, associated with a significant (chi(2)(1df)=4.54, P=0.033) bias in transmission of the haplotype insTT T T T and less (chi(2)(1df)=5.59, P=0.018) than expected transmission of insTT C C C. When compared with data on malaria associations from Gambia, the results suggest a complex pattern of haplotypic variation at the IFNGR1 promoter locus associated with different infectious disease in African populations that reflect the complex roles of IFN-gamma in parasite killing versus inflammation and pathogenesis. PMID- 17136126 TI - Organ donation in the United Kingdom. PMID- 17136127 TI - Indinavir crystalluria. PMID- 17136128 TI - Retroperitoneal fibrosis. PMID- 17136129 TI - Shiga toxin 'goes retro' in human primary kidney cells. AB - The pathway and the efficiency of intracellular trafficking of Shiga toxin differ between cell types, and this impacts on susceptibility to cytotoxicity. Warnier et al. demonstrate that in cell types targeted during human disease, Shiga toxin undergoes retrograde transport via the trans-Golgi network to the endoplasmic reticulum, albeit less efficiently than in HeLa cells. PMID- 17136130 TI - Comprehensive suppression of the renin-angiotensin-aldosterone system in chronic kidney disease: covering all of the bases. AB - Reducing proteinuria and blood pressure in chronic kidney disease (CKD) decreases rate of progression. Inhibition of the renin-angiotensin system by angiotensin converting enzyme inhibitors and/or angiotensin receptor blockers is beneficial in reducing proteinuria but incomplete in suppressing aldosterone production and its renal effects. Adding aldosterone receptor blockers to these other agents may further halt the progression of CKD. PMID- 17136131 TI - Asymmetric dimethylarginine in end-stage renal disease patients: a biomarker modifiable by calcium blockade and angiotensin II antagonism? AB - The nitric oxide synthase inhibitor asymmetric dimethylarginine (ADMA) is an emerging risk biomarker in cardiovascular and renal diseases. Apparently amlodipine and valsartan produce substantial reductions in the plasma concentration of this methylarginine in hemodialysis patients. These findings are of relevance for designing studies aimed at testing the etiologic relevance of ADMA to the high cardiovascular risk of ESRD. PMID- 17136136 TI - Preparing for the inevitable--an influenza pandemic. PMID- 17136137 TI - An introduction to avian and pandemic influenza. AB - There are many types of influenza viruses, which cause illness in a variety of birds and mammals. New strains are constantly evolving, causing seasonal influenza epidemics in humans. This article provides information about influenza and influenza viruses, and the three influenza pandemics of the twentieth century. Pandemic influenza is differentiated from avian influenza, which is a viral disease that primarily infects birds. The current outbreak of avian influenza H5N1 in poultry flocks across the world is unprecedented in its spread. Human infection with avian influenza is rare and for most strains the symptoms are usually mild. A notable exception is HN51, where almost 60 per cent of the currently recorded 251 human cases have died. While the risk of a pandemic occurring in the current circumstances is unknown, there is a high level of concern worldwide. PMID- 17136138 TI - An Australian perspective of the 1918-1919 influenza pandemic. AB - The 1918-1919 influenza pandemic stands as one of the greatest natural disasters of all time. In a little over a year the disease affected hundreds of millions of people and killed between 50 and 100 million. When the disease finally reached Australia in 1919 it caused more than 12,000 deaths. While the death rate was lower than in many other countries, the pandemic was a major demographic and social tragedy, affecting the lives of millions of Australians. This paper briefly assesses the impact of the pandemic on Australia and NSW with particular reference to the demographic and social impact and the measures advanced to contain it. PMID- 17136139 TI - The risk of avian influenza in birds in Australia. AB - Avian influenza viruses may cause mild or severe disease in birds. There have been five recorded outbreaks of highly pathogenic avian influenza in birds in Australia, all of which were caused by the H7 subtype. These were quickly controlled and similar coordinated responses are expected for possible future outbreaks of avian influenza. Migratory birds are not regarded as the source of these outbreaks, and the prevalence of avian influenza viruses in wild birds in Australia is very low. Avian influenza H5N1, which emerged in birds in China in 1996, has spread to bird flocks in Asia, Europe and Africa. The main carriers of avian influenza, ducks, do not migrate to Australia, and currently the risk of H5N1 occurring in Australian birds appears to be low. Nevertheless, surveillance and response plans for outbreaks of highly pathogenic avian influenza have recently been upgraded across Australia. PMID- 17136140 TI - Commonwealth pandemic preparedness plans. AB - This paper describes the work being done by the Australian Government Department of Health and Ageing to prepare for a possible influenza pandemic. It provides an overview of Australian Government initiatives and explains both the purpose and the content of the Australian Health Management Plan for Pandemic Influenza. It summarises efforts to improve regional detection and response capabilities and it explains the role of simulation exercises in Australian pandemic planning. PMID- 17136141 TI - Preparing for the next influenza pandemic: a New South Wales perspective. AB - Pandemic influenza is one of a small number of infectious diseases that pose a significant global threat. Pandemic preparedness has accelerated around the world in recent years in response to the perceived increased risk of a pandemic developing following the emergence of H5N1 avian influenza in domestic poultry flocks in Asia, Africa and Europe. There is a hierarchy of pandemic plans - international, national, state, and local - and harmonisation of all of these is imperative for a coordinated and effective response. At the national and state levels, plans have been developed for a whole-of-government response to a pandemic, in addition to plans specifically for the health sector. It is inevitable that influenza pandemics will occur and careful planning is crucial to mitigate their potentially devastating effects. PMID- 17136142 TI - Pandemic planning at the coal face: responsibilities of the public health unit. AB - Responding to an infectious disease pandemic requires a coordinated approach from all essential services. Public health units across NSW will play an important role in a range of control activities. These include: surveillance, education, communication, case ascertainment, case management (excluding clinical management), infection control, contact tracing, monitoring contacts in home quarantine, surveillance at borders, epidemiological studies and immunisation. Public health units are currently planning for such an emergency and these plans will need to be tested and refined under simulated conditions. PMID- 17136143 TI - Communicable Diseases report, New South Wales, for May and June 2006. PMID- 17136144 TI - Development of a competency-based assessment process for advanced pharmacy practice experiences. AB - OBJECTIVES: To develop and implement a competency-based assessment process for the experiential component of a pharmacy education curriculum. DESIGN: A consultative process was used in the development of new assessment forms and policies, and a survey regarding student and faculty satisfaction was conducted. Information received from the survey and from consultations with faculty preceptors resulted in revision of the forms in subsequent years. ASSESSMENT: Faculty and student perceptions of the assessment process were generally positive. We were moderately successful in reducing grade inflation. The new process also provides the school with data that can be used to evaluate the effectiveness of our curriculum in preparing students for practice. CONCLUSIONS: Development and implementation of a competency-based assessment process require a considerable amount of work from dedicated faculty members. With health professions schools under pressure to provide evidence of their graduates' clinical competence, this is a worthwhile investment. PMID- 17136145 TI - Service scripts: a tool for teaching pharmacy students how to handle common practice situations. AB - OBJECTIVES: This paper describes the use of service scripts to teach pharmacy students how to manage specific practice situations by learning and following scripted behaviors. DESIGN: Based upon role theory, service scripts require specific behaviors for a broad range of practice problems and communicate consistent messages about the responsibilities of all people involved. Service scripts are developed by (1) identifying scenarios for the script, (2) eliciting the script's structure and content, and (3) documenting the reasoning behind the steps in the script. ASSESSMENT: Students in a nontraditional doctor of pharmacy program developed scripts for their practice settings. They concluded that scripts were useful for quickly learning new, routine tasks, but expressed concern that scripts could be misused by pharmacists and managers. The process of script development itself was useful in gaining feedback about common practice problems. CONCLUSION: By mastering managerial, clinical, and communication scripts, students can develop capabilities to provide professional services. PMID- 17136146 TI - A renal transplantation advanced pharmacy practice experience. AB - OBJECTIVES: To establish and evaluate an ambulatory care renal transplantation clinic advanced pharmacy practice experience (APPE). DESIGN: Students spend 5 weeks performing pharmaceutical care activities for renal transplant patients, presenting health-related topics, and conducting research. A paired t test was used to determine differences between students' pre- and post-APPE test scores. Standardized evaluations completed by the preceptor and the students were used to evaluate learning and the APPE. ASSESSMENT: Posttest scores were significantly higher than pretest scores (n = 17; 88.2 +/- 7.3 vs 55.9 +/- 22.4; p < 0.001). Overall, students found this APPE enjoyable and believed that it increased their knowledge concerning transplant medicine and patient care. CONCLUSION: With the recommendation that all transplant programs have clinical pharmacy services, it is imperative to train students to care for transplant patients. Information in this manuscript can be used as a guide for utilizing the combined resources from schools of pharmacy and transplantation centers to implement a renal transplant ambulatory care APPE. PMID- 17136147 TI - The use of wireless laptop computers for computer-assisted learning in pharmacokinetics. AB - OBJECTIVE: To implement computer-assisted learning workshops into pharmacokinetics courses in a doctor of pharmacy (PharmD) program. DESIGN: Workshops were designed for students to utilize computer software programs on laptop computers to build pharmacokinetic models to predict drug concentrations resulting from various dosage regimens. In addition, students were able to visualize through graphing programs how altering different parameters changed drug concentration-time curves. Surveys were conducted to measure students' attitudes toward computer technology before and after implementation. Finally, traditional examinations were used to evaluate student learning. ASSESSMENT: Doctor of pharmacy students responded favorably to the use of wireless laptop computers in problem-based pharmacokinetic workshops. Eighty-eight percent (n = 61/69) and 82% (n = 55/67) of PharmD students completed surveys before and after computer implementation, respectively. Prior to implementation, 95% of students agreed that computers would enhance learning in pharmacokinetics. After implementation, 98% of students strongly agreed (p < 0.05) that computers enhanced learning. Examination results were significantly higher after computer implementation (89% with computers vs. 84% without computers; p = 0.01). CONCLUSION: Implementation of wireless laptop computers in a pharmacokinetic course enabled students to construct their own pharmacokinetic models that could respond to changing parameters. Students had greater comprehension and were better able to interpret results and provide appropriate recommendations. Computer-assisted pharmacokinetic techniques can be powerful tools when making decisions about drug therapy. PMID- 17136148 TI - Creation of a graduate oral/written communication skills course. AB - OBJECTIVE: To convert a traditional graduate seminar course into a class that emphasizes written as well as oral communication skills. DESIGN: Graduate pharmacology/toxicology students presented formal and informal seminars on their research progress and on recent peer-reviewed literature from the field. Students in the audience wrote critiques of the research project or article, as well as of the presentations themselves. ASSESSMENT: Students were evaluated based on oral presentations, class participation, and a scientific writing component. All faculty members provided constructive written comments and a grade. The course master provided the presenter with a formal written review and returned a "red pen" revision of each student critique. CONCLUSION: This novel seminar/writing course introduces intensive focus on writing skills, which are especially essential today given the large number of graduate students for whom English is not a first language. PMID- 17136149 TI - Emotional intelligence instruction in a pharmacy communications course. AB - OBJECTIVES: To determine the benefits of incorporating emotional intelligence instruction into a required pharmacy communications course. DESIGN: Specific learning objectives were developed based upon the emotional intelligence framework and how it can be applied to pharmacy practice. Qualitative data on student perceptions were collected and analyzed using theme analysis. ASSESSMENT: Students found instruction on emotional intelligence to be a positive experience. Students reported learning the taxonomy of emotional intelligence--a concept that previously was difficult for them to articulate or describe, and could use this knowledge in future pharmacy management situations. Students also recognized that their new knowledge of emotional intelligence would lead to better patient outcomes. CONCLUSION: Students had positive perceptions of the importance of emotional intelligence. They valued its inclusion in the pharmacy curriculum and saw practical applications of emotional intelligence to the practice of pharmacy. PMID- 17136150 TI - Medication management in primary and secondary schools. AB - OBJECTIVES: To identify whether and how pharmacy faculty members are addressing the issue of medication management in primary or secondary schools in their teaching, research, and service activities, and to ascertain the extent to which they think the issue is an important one. METHODS: Four hundred ninety-nine faculty members completed a questionnaire inquiring about the research, teaching, and service activities in which they participated that related to medication management in schools. RESULTS: Only 33 subjects (6.6%) addressed the topic of medication management in schools in their courses; only 13 (2.6%) conducted research on the topic; and only 30 (6%) were involved in service in this area. On the other hand, 432 respondents (86.6%) believed that the issue of medication management in schools was either somewhat or extremely important. CONCLUSIONS: There is a large gap between the number of subjects that think medication management in schools is an important topic and the number who actually include the topic in teaching, research, and or service. PMID- 17136151 TI - Programmatic curricular outcomes assessment at colleges and schools of pharmacy in the United States and Puerto Rico. AB - OBJECTIVES: To categorize the manner in which programmatic curricular outcomes assessment is accomplished, identify the types of assessment methodologies used, and identify the persons or groups responsible for assessment. METHODS: A self administered questionnaire was mailed to 89 institutions throughout the United States and Puerto Rico. RESULTS: Sixty-eight of 89 surveys (76%) were returned. Forty-one respondents (60%) had a written and approved plan for programmatic curricular outcomes assessment, 18% assessed the entire curriculum, and 57% had partial activities in place. Various standardized and institution-specific assessment instruments were employed. Institutions differed as to whether an individual or a committee had overall responsibility for assessment. CONCLUSION: To move the assessment process forward, each college and school should identify a person or group to lead the effort. Additional validated assessment instruments might aid programmatic assessment. Future studies should identify the reasons for selecting certain assessment instruments and should attempt to identify the most useful ones. PMID- 17136152 TI - Student perspectives on pharmacy curriculum and instruction in Egyptian schools. AB - OBJECTIVES: To determine student attitudes and opinions towards pharmacy education in Egyptian universities to provide information for designing delivery of a revised pharmacy curriculum. METHODS: Students were recruited from the pharmacy faculties at a government-sponsored university and a privately funded university. Data were gathered using a structured questionnaire and statistically analyzed. Responses from open questions were subjected to thematic analysis. RESULTS: Students spent widely differing amounts of time on non-classroom study, little of which was self-directed. This was reflected in the low frequency of use of library facilities and the preference of students for passively acquired information. Themes that emerged on how students would improve the curriculum were to increase the use of computers and the Internet; make the course more relevant to pharmacy practice and/or clinical pharmacy; improve and expand the practical components of the course; increase their own involvement in learning; and increase their understanding of subjects as well as their knowledge. For many of the questions, there was a significant different between the responses of students at the 2 universities. CONCLUSIONS: Students relied on classroom teaching and devoted little time to self-directed study. However, students were aware of international developments in pharmacy education and practice and are receptive to change. PMID- 17136153 TI - Comparison of students' performance in and satisfaction with a clinical pharmacokinetics course delivered live and by interactive videoconferencing. AB - OBJECTIVES: To compare students' performance in and course evaluations for a clinical pharmacokinetics course taught in a traditional classroom setting, and for the same course taught via interactive videoconferencing. METHODS: The course was taught in a traditional classroom setting to 38 students, and in asynchronous sessions via interactive videoconferencing to 75 students at a distant site. A course evaluation was administered to each group at the conclusion of the courses. RESULTS: The students in the live classroom setting had a higher mean final course grade of 90.7% compared to the mean final course grade (87.8%) of students in the interactive videoconferencing group (P = 0.024). The mean evaluation score for students in the videoconferencing class were higher than for students in the live classroom setting (4.73 vs. 4.58; P < 0.001). CONCLUSIONS: Students in both the classroom setting and interactive videoconferencing setting performed well and had a high overall perception of the course. PMID- 17136154 TI - A comparison of the acceptability and effectiveness of two methods of distance education: CD-ROM and audio teleconferencing. AB - OBJECTIVES: To fulfill a need for convenient and effective continuing education for pharmacists throughout Wisconsin by developing a pharmacy continuing education program using 2 different methods for distance education: audio teleconferencing and CD-ROM. METHODS: Eighty pharmacists were recruited from a 2003 University of Wisconsin distance education program, 47 of whom participated in the course using the traditional audio teleconference method, and 33 of whom participated using a home study CD-ROM containing the same material presented in the teleconference. Volunteers were required to complete a pretest, a first posttest immediately following completion of the continuing education course, a second posttest 1 month following the conclusion of the course, and an evaluation. RESULTS: The CD-ROMs were a more acceptable method for distance education than audio teleconferencing and resulted in better retention of the course information. CONCLUSION: These study results can help coordinators of continuing pharmaceutical education programs to determine the most effective and acceptable method for future distance-education opportunities. PMID- 17136155 TI - Global public health and the academy. PMID- 17136156 TI - Preparing our graduates for a lifetime of learning. PMID- 17136157 TI - The elephant in the room: NIH funding of faculty-initiated award proposals. PMID- 17136158 TI - A career in academic pharmacy: opportunities, challenges, and rewards. PMID- 17136159 TI - Pharmacy education in Jordan, Saudi Arabia, and Kuwait. AB - The practice of pharmacy, as well as pharmacy education, varies significantly throughout the world. In Jordan, Kuwait, and Saudi Arabia, the profession of pharmacy appears to be on the ascendance. This is demonstrated by an increase in the number of pharmacy schools and the number of pharmacy graduates from pharmacy programs. One of the reasons pharmacy is on the ascendance in these countries is government commitment to fund and support competitive, well-run pharmacy programs. In this report we describe pharmacy education in 3 Middle East countries: Jordan, Kuwait, and Saudi Arabia. All 3 countries offer bachelor of pharmacy (BPharm) degrees. In addition, 2 universities in Jordan and 1 in Saudi Arabia offer PharmD degree programs. The teaching methods in all 3 countries combine traditional didactic lecturing and problem-based learning. Faculties of pharmacy in all 3 countries are well staffed and offer competitive remuneration. All 3 countries have a policy of providing scholarships to local students for postgraduate training abroad. The majority of students in Jordan and Kuwait are female, while the ratio of male to female students in Saudi Arabia is even. Students' attitudes towards learning are generally positive in all 3 countries. In Saudi Arabia and Kuwait, most pharmacy graduates work in the public sector, while in Jordan, the majority work in the private sector. PMID- 17136160 TI - Experiential education in community pharmacy. PMID- 17136161 TI - Educating students for practice: educational outcomes and community experience. AB - The education of pharmacists in the United States integrates classroom and experiential learning. Two organizations played a key role in determining the current education of pharmacy students. They are the Accreditation Council for Pharmacy Education and the American Association of Colleges of Pharmacy. The curriculum offered today provides opportunities for students to learn and achieve ability-based outcomes in both didactic and experiential courses. This review of pharmacy education focuses generally on the national leadership of pharmacy education both past and present and specifically on outcomes of practice that students are expected to achieve. Included in the discussion are recommendations for how preceptors in a community practice model can build on the college curriculum by recognizing and incorporating ability-based outcomes into their activities of the introductory and advanced practice courses. PMID- 17136162 TI - Enhancing community pharmacy through advanced pharmacy practice experiences. AB - The pressures driving the need for an expanded practice scope in community pharmacy have been building for the past 2 decades. Many pharmacists have chosen to embrace the pharmaceutical care model in their practice sites to meet patient and healthcare system needs. The potential for medication therapy management (MTM) services provide an additional career opportunity for pharmacy graduates. Colleges of pharmacy offer advanced pharmacy practice experiences (APPEs) in the community setting that are designed to prepare students for these opportunities. These sites provide students with the opportunity to observe the integration of pharmaceutical care activities into community practice. Although developing an APPE site is challenging, serving as a preceptor benefits the students, the site, and the patients served. Therefore, colleges of pharmacy and community pharmacists are collaborating to increase the number of APPE sites to prepare pharmacy students for practice today and tomorrow. PMID- 17136163 TI - Organizing a community advanced pharmacy practice experience. AB - Setting up a community advanced pharmacy practice experience can be an overwhelming task for many pharmacy preceptors. This article provides guidance to pharmacist preceptors in developing a complete and effective community advanced pharmacy practice experience (APPE). When preparing for the APPE, initial discussions with the college or school of pharmacy are key. Benefits, training, and requirements should be addressed. Site preparation, including staff education, will assist in the development process. The preceptor should plan orientation day activities and determine appropriate evaluation and feedback methods. With thorough preparation, the APPE will be rewarding for both the student and the pharmacy site. PMID- 17136164 TI - Developing structured-learning exercises for a community advanced pharmacy practice experience. AB - The recent growth in the number of pharmacy schools across the nation has resulted in the need for high-quality community advanced pharmacy practice experience (APPE) sites. A vital part of a student's education, these APPEs should be structured and formalized to provide an environment conducive to student learning. This paper discusses how to use a calendar, structured-learning activities, and scheduled evaluations to develop students' knowledge, skills, and abilities in a community pharmacy setting. PMID- 17136167 TI - Professional practice plans: recommendations from the 2005 Council of Faculties Council of Deans Task Force. AB - OBJECTIVES: Determine the degree to which AACP member schools have established professional practice plans, characterize the nature of existing practice plans, and provide recommendations on the implementation of practice plans at AACP member schools. DESIGN: Survey of CEO Deans of AACP member institutions administered via online survey instrument. RESULTS: Sixty-five schools responded, with 29 (45%) indicating that they had an active practice plan in place. Fifty two percent of those who do no have practice plans in place anticipate having plans established within three years. A variety of revenue sources are addressed by existing practice plans including sponsored research, patient care, educational activities and consulting. CONCLUSIONS: Academic pharmacy lacks sophistication in regards to developing comprehensive professional practice plans. Colleges of pharmacy should consider differentiating plans that address monies collected from sponsored research vs. professional services. AACP should continue to monitor this topic as increasing participation by member schools is expected. PMID- 17136168 TI - The pharmacy student population: applications received 2004-05, degrees conferred 2004-05, fall 2005 enrollments. PMID- 17136169 TI - Pharmacy student response to patient-simulation mannequins to teach performance based pharmacotherapeutics. AB - OBJECTIVE: To design and implement a simulated patient-case assessment using a mannequin for critical care pharmacotherapeutic education of doctor of pharmacy students and to evaluate student satisfaction with the simulation. DESIGN: During the second year of the doctor of pharmacy program, all students were required to complete Introduction to Critical Care. This course consisted of didactic education, written patient-case sessions, and an interactive patient simulation session. Information on the patient case was distributed to students after completing the didactic portion of the course. Patient information was programmed into a simulation mannequin, which demonstrated characteristics of a critically ill human. Students were surveyed post-simulation to determine the effectiveness of the learning experience. ASSESSMENT: The majority of students (88%) were extremely satisfied with the experience. The facilitator was considered to be extremely useful in 75% of responses. CONCLUSION: By simulating a patient case, the facilitator was able to control students' learning environment, adapt the simulation to the level of the students' performance, and debrief students immediately. Ultimately, by involving students in actual patient cases early in the pharmacy curriculum, this type of education could produce pharmacists with a high level of expertise and confidence. PMID- 17136170 TI - Evaluation of pharmaceutical care opportunities within an advanced pharmacy practice experience. AB - OBJECTIVE: To determine the nature and extent of learning opportunities presented to students during a community advanced pharmacy practice experience. METHODS: Learning opportunities were determined by reviewing students' portfolios. The numbers of unique learning opportunities in which students were engaged were calculated. RESULTS: While all students had ample opportunities to participate in consultations involving the assessment of new and refill prescriptions and nonprescription drugs, only 78% of students had the opportunity to engage in even 1 comprehensive pharmaceutical care consult over an 8-week period. Students' experiences in providing follow-up care, participating in a patient care project, and shadowing another healthcare professional were also limited. CONCLUSION: Analysis of the student portfolios suggested that the learning environment did not provide students with adequate opportunities to develop pharmaceutical care competencies. PMID- 17136171 TI - Influence of attitudes toward curriculum on dishonest academic behavior. AB - OBJECTIVES: The objective of this study was to examine possible associations between students' self-reported behaviors and opinions towards academic dishonesty, and their attitudes towards curriculum, assessment, and teaching within the pharmacy program. METHODS: A questionnaire was developed and distributed to undergraduate (pre-licensure) students at 4 schools of pharmacy in Canada, including students enrolled in the international pharmacy graduate program. RESULTS: More than 80% of respondents indicated they had participated in one or more of the act of academic dishonesty described in the questionnaire. A weak to moderate correlation was found between students' attitudes towards pharmacy education and their self-reported behaviors related to academic dishonesty. CONCLUSIONS: This study confirmed previous findings suggesting widespread academic dishonesty as well as a hierarchy of values with respect to students' perceptions regarding severity and importance of academic dishonesty. Despite methodological limitations inherent in examining academic dishonesty, there is a definite need to continue to examine this important issue. While this study indicated only a moderate correlation between attitudes towards curriculum and dishonest behaviors, the problem of academic misconduct is multifactorial and will require ongoing study. PMID- 17136172 TI - Drug information education in doctor of pharmacy programs. AB - OBJECTIVE: To characterize pharmacy program standards and trends in drug information education. METHODS: A questionnaire containing 34 questions addressing general demographic characteristics, organization, and content of drug information education was distributed to 86 colleges and schools of pharmacy in the United States using a Web-based survey system. RESULTS: Sixty colleges responded (73% response rate). All colleges offered a campus-based 6-year first professional degree PharmD program. Didactic drug information was a required course in over 70% of these schools. Only 51 of the 60 colleges offered an advanced pharmacy practice experience (APPE) in drug information, and 62% of these did so only on an elective basis. CONCLUSION: Although almost all of the PharmD programs in the US include a required course in drug information, the majority do not have a required APPE in this important area. PMID- 17136173 TI - Weekly active-learning activities in a drug information and literature evaluation course. AB - OBJECTIVES: To incorporate learning activities into the weekly 2-hour Drug Information and Literature Evaluation class sessions to improve student ability and confidence in performing course objectives, as well as to assess student perception of the value of these activities. METHODS: In-class activities that emphasized content and skills taught within class periods were created and implemented. Three different surveys assessing student ability and confidence in completing drug information and literature retrieval and evaluation tasks were administered prior to and following the appropriate class sessions. At the completion of the course, an additional evaluation was administered to assess the students' impressions of the value of the learning activities. RESULTS: Students reported increased ability and confidence in all course objectives. The teaching activities were also stated to be useful in students' learning of the material. CONCLUSIONS: Incorporation of weekly learning activities resulted in an improvement in student ability and confidence to perform course objectives. Students considered these activities to be beneficial and to contribute to the completion of course objectives. PMID- 17136174 TI - A predictive validity study of the Pharmacy College Admission Test. AB - OBJECTIVE: To examine the validity of Pharmacy College Admission Test (PCAT) scores for predicting grade point averages (GPAs) of students in years 1-4 of pharmacy programs. METHODS: Data were collected from 11 colleges and schools of pharmacy: entering cumulative and math/science GPAs, PCAT scaled scores, pharmacy program GPAs for years 1-4, student status after 4 years. Correlation, regression, discriminant, and diagnostic accuracy analyses were used to determine the validity of the PCAT for predicting subsequent GPAs. RESULTS: PCAT scaled scores and entering GPAs were positively correlated with subsequent GPAs. Regression analyses showed the predictive value of the PCAT scores, especially in combination with entering GPAs. Discriminant and diagnostic accuracy analyses supported these findings and provided practical suggestions regarding optimal PCAT scores for identifying students most likely to succeed. CONCLUSION: Both PCAT scaled scores and entering cumulative GPAs showed moderate to strong predictive validity as indicators of candidates likely to succeed in pharmacy school. PMID- 17136175 TI - Student-directed learning in a community geriatrics advanced pharmacy practice experience. AB - OBJECTIVES: To assess a community geriatrics advanced pharmacy practice experience (APPE) that aimed to improve students' attitudes towards older adults and provide a student-directed learning experience. DESIGN: Students provided blood pressure monitoring and medication counseling to older adults living in a low-income residential facility as part of a required 6-week ambulatory care service-learning APPE. Pre-experience and post-experience essays on students' perceptions of the elderly and their intended and actual learning were retrospectively reviewed using a qualitative process to determine whether the course objectives were met. ASSESSMENT: Many students initially described older adults in factual terms or using negative descriptors. Most expressed a desire to increase their knowledge of diseases commonly occurring in and drugs commonly prescribed for the elderly or to improve specific skills. Many students initially had difficulty articulating clear and measurable learning objectives and appropriate assessment metrics, which are important components of self-directed learning. The final essays revealed many students learned more about the humanistic aspects of care than they had anticipated. CONCLUSION: This community based geriatrics experience improved students' attitudes towards working with older adults and provided practice in developing and assessing their personal learning objectives. PMID- 17136176 TI - An elective course in cultural competence for healthcare professionals. AB - OBJECTIVES: To develop and assess a cultural competence elective course used to increase students' levels of awareness, acceptance, and understanding of the role of culture on healthcare perceptions. DESIGN: Classroom and at-home assignments were designed, small group discussions were conducted, and guest speakers were recruited to allow students to explore different viewpoints about other cultures. ASSESSMENT: Student performance was assessed based on assignments and participation in class discussions. Student satisfaction with the course was assessed using a standard course evaluation form. CONCLUSION: Participation in the course resulted in greater student understanding of cultural issues facing healthcare providers and in students feeling more capable of integrating cultural factors into patient care. PMID- 17136177 TI - Evaluation of structured patient interactions: the Diabetes Check. AB - OBJECTIVES: To evaluate a structured patient intervention, the Diabetes Check, in which pharmacy students ask patients with diabetes about 3 clinical targets: A1c, blood pressure, and low-density lipoprotein (LDL) cholesterol (ABC) levels. Specific objectives were to: (1) describe the effect of patients' reactions on pharmacy students; (2) describe pharmacy students' confidence and role beliefs; and (3) determine predictors of pharmacy students' intention to use the Diabetes Check in the future. METHODS: After training, pharmacy students were asked to perform 10 diabetes checks and complete an evaluation. Data from the evaluations described pharmacy students' reactions, role beliefs, and confidence. Linear regression was performed to predict intention of using the Diabetes Check in future. RESULTS: One-hundred twenty-eight pharmacy students used the Diabetes Check tool with over 1000 patients. Most pharmacy students were encouraged by patients' reactions (56%). Pharmacy students' perceptions of patients' reaction and pharmacy students' role beliefs about monitoring ABCs significantly predicted pharmacy students' reported intention to use a Diabetes Check (r-squared = 0.52). CONCLUSIONS: Pharmacy students' perceptions of patient reactions and role beliefs about the importance of monitoring predicted their reported intention of performing a Diabetes Check in the future. PMID- 17136178 TI - Obtaining and providing health information in the community pharmacy setting. AB - Community pharmacists are a valuable information resource for patients and other healthcare providers. The advent of new information technology, most notably the Internet, coupled with the rapid availability of new healthcare information, has fueled this demand. Pharmacy students must receive training that enables them to meet this need. Community advanced pharmacy practice experiences (APPEs) provide an excellent opportunity for students to develop and master drug information skills in a real-world setting. Preceptors must ensure that students are familiar with drug information resources and can efficiently identify the most useful resource for a given topic. Students must also be trained to assess the quality of resources and use this information to effectively respond to drug or health information inquiries. This article will discuss key aspects of providing drug information in the community pharmacy setting and can serve as a guide and resource for APPE preceptors. PMID- 17136179 TI - Improving communication skills of pharmacy students through effective precepting. AB - Pharmacy students should be given opportunities to learn and practice interpersonal communication skills during their community advanced pharmacy practice experience (APPE). Preceptors have the responsibility of setting the stage for the pharmacy students during their initial encounter. During this orientation to the site, students should become familiar with the history of the practice, the types of services provided, and the staff members. Once the orientation is completed, preceptors can develop strategies for incorporating the students into the practice's patient care activities. Students should participate in patient counseling, interviewing, and educational sessions. Also, students should participate in collaborative work with other health care providers. To ensure the development of communication skills in pharmacy students, preceptors can incorporate the teaching process "see one, do one, teach one" into their teaching activities. By following these strategies, preceptors can effectively and positively impact the communication skills of their students. PMID- 17136180 TI - Improving student professionalism during experiential learning. AB - The purpose of this paper is to serve as a tool for preceptors to aid in pharmacy students' development of professionalism. Specifically, the article defines professionalism, describes it in the context of contemporary pharmacy practice, discusses the professional socialization process of students, and suggests strategies for preceptors to facilitate improvement in professionalism among students during experiential training. While numerous suggestions are presented, positive role modeling is considered the most important means of improving professionalism among students. PMID- 17136181 TI - An assessment system for mapping CAPE outcomes in an advanced pharmacy practice experience program. AB - OBJECTIVE: To implement and evaluate an assessment system based on the 1998 Center for the Advancement of Pharmaceutical Education's (CAPE) Outcomes for students in advanced pharmacy practice experiences (APPEs). DESCRIPTION: The system requires each preceptor to create a summative assessment tool by choosing the most important 20-30 CAPE competencies and sub-elements necessary for his/her pharmacy practice with each to be scored by him/her on a 4-point scale from "exceeds expectations" to "below expectations." Students' grades are determined by an examination committee based on the preceptors' assessments. The system contains a mechanism to assist students with competency deficits and permits a student's yearlong performance in the APPE program to be considered when assigning grades for individual APPE courses. EVALUATION: The assessment system permits each student's performance in individual APPE courses to be mapped to the CAPE competencies for grading purposes. It permits class performance in an APPE program to be mapped to the CAPE competencies for quality assurance and school planning. CONCLUSION: An assessment system based on the 1998 CAPE Educational Outcomes competencies has been successfully introduced for students in APPE training. PMID- 17136184 TI - Strength-based advocacy: making a difference through teaching, research, and service. PMID- 17136185 TI - Self-assessment in pharmacy education. PMID- 17136186 TI - Evaluation of evidence for interprofessional education. AB - Based on recommendations from numerous organizations, educators in healthcare disciplines are implementing interprofessional training programs. Our objective was to summarize relevant literature in a way that would be most useful to clinician educators. Studies involving educational interventions in health professions to enhance learner-based outcomes relevant to the provision of interprofessional care were identified. We sought prospective, controlled trials in which at least 2 health care disciplines were represented, and 1 of which was medicine. Thirteen reports met the criteria for inclusion. Interventions varied widely in design and intensity, but generally included both didactic and clinical components and lasted several weeks or longer. Most studies used pretest/posttest controls and observed positive effects on learners' attitudes and knowledge. Combined clinical and didactic experiences may produce short-term improvements in learners' knowledge and attitudes about interprofessional care. Future research should employ control groups and validated, behaviorally oriented outcome measures whenever possible. PMID- 17136187 TI - Communicating with patients: skills assessment in US colleges of pharmacy. AB - OBJECTIVE: To describe current practices in assessing patient communication skills in US colleges and schools of pharmacy. METHODS: Syllabi and behavioral assessment forms were solicited and key faculty members were interviewed. Forms were analyzed to determine skills most commonly assessed in communication with simulated or role-playing patients. RESULTS: Fifty schools submitted behavioral assessment forms for patient communication skills. Individuals from 47 schools were interviewed. Colleges were found to vary in the way communication skills were assessed. Assessment forms focused more on dispensing a new prescription than monitoring ongoing therapy. Providing information was emphasized more than promoting adherence. Common faculty concerns were lack of continuity and congruence of assessment across the curriculum. CONCLUSIONS: A common understanding of the standards and procedures for determining competence is needed. Experience and assessment activities should be sequenced throughout a program to build competence. PMID- 17136188 TI - Advanced practice internship: experiential learning in a drug use and disease state management program. AB - OBJECTIVE: Establish a 3-year hospital internship within a drug use and disease state management program that would provide doctor of pharmacy students with experiential learning while still completing their classroom studies. DESIGN: As paid interns, students engaged in group and individual activities that assessed clinical practice guidelines. Patient monitoring and clinical intervention techniques were learned through prospective evaluation of drug therapy. Students designed evidence-based treatment guidelines and participated in all phases of development, including multidisciplinary approval, implementation, and evaluation stages. ASSESSMENT: Student competency was continually monitored through direct observation by a preceptor and written examinations. Patient case studies, group discussions, and poster presentations allowed assessment of student growth in knowledge and communication skills. CONCLUSION: The comprehensive structure of this internship provides a broad perspective for understanding the role of the hospital pharmacist in providing pharmaceutical care. Close supervision maximizes student learning potential and fosters a mentoring relationship for both personal and professional growth. PMID- 17136189 TI - Emotional intelligence as a predictor of academic and/or professional success. AB - The concept of "emotional intelligence" has been extensively popularized in the lay press and corporate world as individuals purport the potential ability of emotional intelligence to predict various markers of success. Emotional intelligence (EI) most commonly incorporates concepts of emotional expression and regulation, self-awareness, and empathy. The concept has been criticized by some for its loose definition and parallels to personality traits. Additionally, several limitations to the instruments used to measure emotional intelligence have been identified. This review examines the foundations of the definitions of emotional intelligence as well as existing educational research involving emotional intelligence, both within the health professions and externally. Recommendations for future research and research potential are discussed. PMID- 17136190 TI - Peer tutoring programs in health professions schools. AB - OBJECTIVE: Peer tutoring programs may be one method of maintaining quality of pharmacy education in the face of growing student enrollment and a small faculty body. A critical review of the literature was performed to ascertain whether peer tutoring programs improve or maintain the academic performance of health care professional students. METHODS: Various electronic databases and abstracts from past American Association of Colleges of Pharmacy's annual meetings were searched to identify pertinent research. Only those articles with quantitative data, an experimental design, and comparative statistical analysis were included for review. RESULTS: Most studies found that peer tutoring had a positive impact on academic performance. These results may not be readily generalizable as there were numerous methodological flaws and limited descriptions of the programs and participants. IMPLICATIONS: Studies with better designs and more detail are needed to answer definitively whether peer tutoring is of benefit. Details of what resources were required should be included in the study to allow the reader to determine the feasibility of the intervention. PMID- 17136191 TI - Pharmacy education: a student's perspective. PMID- 17136192 TI - Characteristics, prevalence, attitudes, and perceptions of academic dishonesty among pharmacy students. AB - OBJECTIVES: To ascertain background factors that influence pharmacy students' willingness to cheat, describe attitudes regarding methods of cheating, assess prevalence of cheating and determine atmospheres that may aid in preventing academic dishonesty. METHODS: Third-professional year PharmD students at 4 institutions participated in a survey administered by a class representative. RESULTS: Of the 296 students who completed survey instruments, 16.3% admitted to cheating during pharmacy school. Approximately 74% admitted that either they or their classmates had worked on an individual assignment with a friend. Students who cheated during high school or in a prepharmacy program were more likely to cheat during pharmacy school (p < 0.0001). Those who possessed a bachelor of science (BS) degree prior to pharmacy school were less likely to cheat (p < 0.0001). CONCLUSIONS: Academic dishonesty is prevalent among pharmacy students. While few respondents directly admitted to cheating, many admitted to activities traditionally defined as dishonest. PMID- 17136193 TI - Pharmacy students' learning styles before and after a problem-based learning experience. AB - OBJECTIVES: To determine learning-style scores of second-year pharmacy students before and after a problem-based learning (PBL) teaching experience and to evaluate the relationships between scores and demographic variables. METHODS: The Grasha-Reichmann Student Learning Style Scale (GRSLSS) was given to pharmacotherapy laboratory students before and after a semester-long problem based learning class. RESULTS: Only one of the GRSLSS score variables was reported as "high" ("collaborative") and none were reported as "low." Students' "avoidant" mean score increased and "participant" mean score decreased after completing the PBL experience (p < or = 0.05). CONCLUSION: While PBL appears to be a teaching style that is conducive to the learning preferences of this cohort of pharmacy students, significant changes in learning styles were noted after completing the PBL experience. These changes may reflect difficulties that occurred in adapting from a didactic teaching style to PBL, and specifically, difficulties in adjusting to participating in a group learning experience. A major limitation was that the teaching style of the facilitator was not measured. PMID- 17136194 TI - Pharmacy students' perceptions of tobacco sales in pharmacies and suggested strategies for promoting tobacco-free experiential sites. AB - OBJECTIVE: The objectives of this report were to estimate the extent to which pharmacy student perceptions are aligned with the 2003 resolution of the American Association of Colleges of Pharmacy (AACP) addressing the use of experiential sites that sell cigarettes and other tobacco products. METHODS: Pharmacy students participating in a national tobacco cessation training program completed posttraining survey instruments and indicated their opinion about tobacco sales in pharmacies. Responses were examined with respect to students' sex and tobacco use status. RESULTS: Of 3,064 students, 3.5% were in favor of tobacco sales in pharmacies. Opinions varied by students' sex (p < 0.001) and tobacco use status (p < 0.001); in logistic regression analyses, males (OR = 2.62; 95% CI, 1.77, 3.89) and current tobacco users (OR = 2.31; 95% CI, 1.41, 3.76) were most likely to be in favor of tobacco sales. CONCLUSION: Few pharmacy students are in favor of tobacco sales in pharmacies. Given the overall lack of support, and acting in accordance with the 2003 AACP resolution, pharmacy schools are encouraged to use only experiential sites that do not sell tobacco products. Suggested strategies for moving toward this goal are presented. PMID- 17136195 TI - A comparison of pharmacy students' confidence and test performance. AB - OBJECTIVES: The objectives of this study were to estimate student retention of knowledge regarding the management of patients with hypertension and dyslipidemia, measure student clinical confidence, and identify the relationship between clinical confidence and actual performance on a knowledge assessment test. METHODS: This was a sequential cross-sectional study to evaluate knowledge retention and clinical confidence of second-year pharmacy students. To measure student clinical confidence, a 12-item clinical confidence questionnaire was administered. To measure student retention of knowledge, a 21-question knowledge assessment test was administered. At least 1 test question was related to each question asked in the clinical confidence questionnaire. RESULTS: One hundred eight students completed the study. The percentage of students correctly answering test questions decreased from a baseline of 70.4% +/- 5.8% to 60.9% +/- 5.8% four months later (p = 0.02) in spite of the students rating their clinical confidence from moderate to high in all areas. The proportion of students answering questions correctly was similar across the different levels of confidence. CONCLUSION: Overall, retention of knowledge appears to decline over a 4-month period of time. Furthermore, while students perceived moderate to high confidence, student knowledge did not match perceived confidence. PMID- 17136196 TI - Pharmacy students' and graduates' attitudes towards people with schizophrenia and severe depression. AB - OBJECTIVES: To compare the attitudes of third-year pharmacy students and pharmacy graduates towards people with schizophrenia and severe depression. METHODS: Third year pharmacy students (n = 216) and pharmacy graduates (n = 232) completed a survey instrument with 21 common items. The third-year students had not yet received any mental health lectures or tutorials as part of their pharmacy course. The graduates had completed their university education, including mental health lectures and tutorials, plus 6 months of supervised clinical practice in the pharmacy workplace. RESULTS: There were no significant differences between the third-year students and pharmacy graduates in terms of social distance from people with schizophrenia. The rates of stigmatization of people with schizophrenia and severe depression were also similar between the groups. CONCLUSIONS: Mental health lectures and tutorials delivered by pharmacists and supervised clinical practice in the pharmacy workplace may not decrease students' social distance or stigmatization of people with mental illness. The results of our study suggest that more comprehensive education and training programs are needed to improve the ability of pharmacists to meet the needs of people with mental illness. PMID- 17136198 TI - Utilization of the Center for the Advancement of Pharmaceutical Education Educational outcomes, revised version 2004: report of the 2005 American College of Clinical Pharmacy Educational Affairs Committee. PMID- 17136199 TI - 2006 Rho Chi Lecture: Unparalleled opportunities for improving medication-related health outcomes. PMID- 17136200 TI - An interdisciplinary approach to introducing professionalism. AB - OBJECTIVE: To provide interdisciplinary structured activities in academic and clinical settings for introducing the concept of professionalism to health professions students. DESIGN: Undergraduate and graduate students from 8 health care disciplines including pharmacy, nursing, communication sciences and disorders, dietetics and nutrition science, genetic counseling, advanced medical imaging, medical technology, and physical therapy participated in an orientation program focusing on interdisciplinary health care and professionalism, as well as a field experience. ASSESSMENT: Survey results from both components (orientation, n = 284; field experience, n = 123) indicated that the project was valuable in increasing students' awareness of (1) the importance of professionalism in the clinical setting and (2) the potential contributions of their profession to the health care team. CONCLUSION: Health professions curricula should include interdisciplinary learning opportunities that enhance collaboration, collegiality, and professionalism among future members of the health care team. PMID- 17136201 TI - The impact of problem-based learning on students' perceptions of preparedness for advanced pharmacy practice experiences. AB - OBJECTIVES: To evaluate graduating pharmacy students' perceptions of their preparedness for advanced pharmacy practice experiences and the effectiveness of problem-based learning in their preparation. METHODS: A survey instrument was administered anonymously in May 2004 and May 2005 to graduating pharmacy students of the University of Mississippi School of Pharmacy. RESULTS: Students reported that the areas in which problem-based learning prepared them most effectively for advanced pharmacy practice experiences were retrieval of medical information (80%), discussion of disease states and drug therapies at the basic science level (56%), and evaluation of the appropriateness of a medication regimen based on patient specific information (50%). Areas in which students reported being inadequately prepared included identifying and utilizing drug assistance programs (42%) and processing prescriptions/hospital orders (40%). CONCLUSION: Data from 2 consecutive graduating classes supports that problem-based learning is an effective format for preparing pharmacy students for advanced pharmacy practice experiences in a variety of areas. PMID- 17136202 TI - Evaluating the accuracy of health news publications in a drug literature evaluation course. AB - OBJECTIVES: To design an assignment for second-professional year pharmacy students to assess the accuracy and quality of health information published in the news. DESIGN: Students in a literature evaluation course were assigned a health-related news publication to review and find the original published research article. They then critically evaluated the quality and accuracy of the news publication based on the original research. All students wrote a critique focusing on the quality and accuracy of the news article and potential responses the lay public might have. ASSESSMENT: Eighty-four percent of students agreed the writing assignment reinforced critical literature evaluation skills, while 90% agreed the assignment contributed to completion of course objectives. CONCLUSIONS: A writing assignment requiring comparison of a news publication to the original research reinforces critical literature evaluation and communication skills, as well as stimulates thought about the accuracy, quality, and public responses to health information published in the news. PMID- 17136203 TI - "Blogging" about course concepts: using technology for reflective journaling in a communications class. AB - OBJECTIVE: Web log technology was applied to a reflective journaling exercise in a communication course during the second-professional year at the University of Mississippi School of Pharmacy, to encourage students to reflect on course concepts and apply them to the environment outside the classroom, and to assess their communication performance. DESIGN: Two Web log entries per week were required for full credit. Web logs were evaluated at three points during the term. At the end of the course, students evaluated the assignment using a 2-page survey instrument. ASSESSMENT: The assignment contributed to student learning and increased awareness level for approximately 40% of the class. Students had few complaints about the logistics of the assignment. CONCLUSION: The Web log technology was a useful tool for reflective journaling in this communications course. Future versions of the assignment will benefit from student feedback from this initial experience. PMID- 17136204 TI - Development of an instrument to measure professionalism. AB - OBJECTIVES: To develop and validate an instrument that measures professionalism among pharmacy students and recent graduates. METHODS: A pharmacy professionalism survey instrument developed by a focus group was pretested and then administered to all first-year pharmacy students enrolled in the University of Georgia College of Pharmacy and to recent pharmacy graduates who were taking the preparation course for the Georgia Pharmacy Law Examination and North American Pharmacist Licensure Examination. Participants were asked to indicate the extent to which they agreed or disagreed with each of 32 items using a 5-point Likert scale. RESULTS: One hundred thirty first-year pharmacy students and 101 pharmacy graduates participated in the survey. Statistical analysis identified 6 factors (subscales), which were later named excellence, respect for others, altruism, duty, accountability, and honor/integrity, the 6 tenets of professionalism. Item to total correlations ranged from 0.25 to 0.57 on the 6 factors (subscales), and reliability estimates ranged from 0.72 to 0.85 for the 6 factors and total scale. CONCLUSIONS: The Pharmacy Professionalism Instrument measures the 6 tenets of professionalism and exhibits satisfactory reliability measures. Future studies using this scale in other pharmacy populations are needed. PMID- 17136205 TI - A global perspective on managed care and pharmacy education. PMID- 17136206 TI - Preparing for the Nexters. PMID- 17136207 TI - Pharmacy students' opinions on civility and preferences regarding professors. AB - OBJECTIVES: To determine the types of classroom behavior that pharmacy students consider uncivil, participation in such behaviors, what type of professor and classroom setting they prefer, and changes in these opinions over time. METHODS: A survey instrument was used to collect data about students' feelings concerning incivility, participation in uncivil behaviors, and preferences concerning classroom experience. Demographic data were used to identify characteristics of student populations and ensure the same students were studied over different time periods. RESULTS: Younger students felt cheating was the most uncivil classroom behavior while older students most disliked cell phone/beeper use. Chewing gum was least offensive for all groups. Students desired that teachers cared about their learning experience, but few would phone a professor at home. CONCLUSIONS: Differences in views concerning civility were found among pharmacy students in their first, third, and fourth years, which may indicate that students' beliefs, actions, and preferences change as they progress through the curriculum. PMID- 17136208 TI - A quantitative assessment of a medicinal chemistry problem-based learning sequence. AB - OBJECTIVES: To establish an aggressive problem-based learning (PBL) format for the medicinal chemistry course and assess the outcomes of student learning. METHODS: To assess learning in the new format, precourse and postcourse examinations were given to students enrolled before and after problem-based learning was implemented, and appropriate statistical analyses were conducted. RESULTS: The PBL cohort did not learn the same amount of factual content yet performed the same on higher-order thought questions as the non-PBL cohort. CONCLUSIONS: Problem-based learning may not be the ideal method for teaching medicinal chemistry. This may be due to several factors including: student learning type, the lack of a cognitive framework for learning in the basic sciences, and time constraints. PMID- 17136209 TI - Using performance-based assessments to evaluate parity between a campus and distance education pathway. AB - OBJECTIVES: To compare the performance of campus-based students with that of distance students during the first 2 years of a doctor of pharmacy program to evaluate parity between the pathways. METHODS: Twelve cases were created for each year of the program along with performance criteria. The cases were converted into computer-based simulations for programmatic assessment at the end of the 2002-2003 and 2003-2004 school years. All first-professional year (P1) and second professional year (P2) students participated in the assessments. Overall class means were calculated and used to compare student performances between campus and distance education pathways. RESULTS: Overall scores for the 2003 P1 class were 56.4% for the campus-based students and 62.4% for the distance students, (p = 0.002); overall scores for the 2003 P2 class were 48.8% and 55.5%, respectively (p < 0.0001). The 2004 overall scores for P1 campus and distance students were 59.0% and 65.7%, respectively, (p = 0.001); and for 2004 P2 scores the results were 51.8% and 56.5%, respectively (p = 0.049). CONCLUSIONS: Students receiving their pharmacy education via a distance pathway scored higher on performance based assessments compared with students receiving their pharmacy education via the traditional campus-based pathway. This indicates that distance students are receiving at least an equivalent curricular experience in the P1 and P2 years compared to that received by campus-based students. PMID- 17136210 TI - Instructional model to teach clinically relevant medicinal chemistry. AB - The relevance of medicinal chemistry to pharmacy practice has been questioned by many pharmacy educators as more emphasis has been placed on linking clinical knowledge and practice to pharmacy student educational outcomes. Faculty teaching in medicinal chemistry and other biomedical and pharmaceutical science courses have embraced this challenge. Various teaching methods and approaches within medicinal chemistry that emphasize application of this knowledge have been sought to improve the usefulness of this scientific discipline to the future careers of students. The newly revised ACPE guidelines and standards have reemphasized the role of the sciences in the curriculum. With this mandate, it is essential that all science faculty members adjust the way they teach to meet the new desired outcomes for pharmacy graduates. This manuscript describes an instructional model for teaching medicinal chemistry explicitly designed to meet these outcomes. A process of collaboration between experienced pharmacy faculty scholars was used to derive this approach. Pedagogy for cognitive and affective learning was incorporated. A case study using a representative drug class is presented to illustrate this model. PMID- 17136211 TI - Count and be counted: preparing future pharmacists to promote a culture of safety. PMID- 17136212 TI - Regarding "Status of PharmD/PhD programs in colleges of pharmacy". PMID- 17136213 TI - Is the thyroid still important in major depression? PMID- 17136214 TI - Cognitive dysfunctions in schizophrenia: potential benefits of cholinesterase inhibitor adjunctive therapy. AB - OBJECTIVE: In schizophrenia, cognitive dysfunctions commonly affect attention, memory and executive function, interfere with functional outcome and remain difficult to treat. Previous studies have implicated the cholinergic system in cognitive functioning. In Alzheimer's disease, cholinergic agonists have shown modest clinical benefits on cognitive and behavioural symptoms. Impaired cholinergic activity might also be involved in schizophrenia. Hence the role of cholinesterase inhibitors (ChEI) as adjunctive therapy is under study. We aimed to review the literature and evaluate the overall effectiveness of ChEI adjunctive therapy for the management of cognitive dysfunctions in schizophrenia. METHODS: We conducted a computer-based search using PubMed (up to February 15, 2006) and ISI Web of Science (conference proceeding abstracts from January 2003 to December 2005) databases. We used the search terms "schizophrenia," "cognition or memory" and "tacrine or donepezil or rivastigmine or galantamine." Studies included were critically analyzed for allocation, blindness, duration and study design, demographic data, and clinical and neuropsychological outcome assessments. We excluded studies that involved patients with psychiatric disorders other than schizophrenia-spectrum or if they involved animals or molecular investigations. We also excluded conference proceeding abstracts with no explicit neuropsychological battery and/or results. RESULTS: Data on ChEI as adjunctive therapy for the cognitive impairments in schizophrenia are sparse and so far derived from small samples and mostly open uncontrolled studies. ChEI's potential in long-term management has barely been documented and remains to be fully explored. CONCLUSION: There is insufficient evidence on whether ChEI should be used for the treatment of cognitive dysfunctions in schizophrenia. Nevertheless, further studies with appropriate trial designs and outcome measures in homogenous schizophrenia populations are warranted. PMID- 17136215 TI - Depression in acute stroke. AB - OBJECTIVE: Depression is one of the most frequent neuropsychiatric disturbances in stroke patients. The clinical aspects and correlations of depression in the first days after acute stroke are less known. This study aimed to 1) assess the frequency of depression, 2) describe the profile of depression of stroke patients and 3) analyze the relation between depression and demographic, predisposing and precipitating conditions, and clinical and imaging data, in acute stroke patients. METHODS: We used the Montgomery-Asberg Depression Rating Scale to assess depression in 178 consecutive acute ( 0.45) of depression between control subjects and acute stroke patients. CONCLUSIONS: Depression was present in almost one-half of the acute stroke patients and was related to previous mood disorder but not not to stroke type or location. Apathy/loss of interest was the predominant clinical feature. PMID- 17136216 TI - Differential effects of naltrexone on cardiac, subjective and behavioural reactions to acute ethanol intoxication. AB - OBJECTIVE: Alcohol may have psychomotor stimulant properties during the rising limb of the blood alcohol curve at commonly self-administered doses. Increased heart rate (HR) immediately after alcohol consumption may serve as an indicator or marker of such properties, which appear to be potentially opiate-mediated and dopamine-dependent. Naltrexone, an opiate antagonist, has been used successfully in the treatment of alcoholism and may produce its therapeutic effects through its effects on alcohol metabolism or by blocking alcohol's rewarding effects. We hypothesized that, if naltrexone blocks the psychomotor stimulant properties of ethanol, then it would decrease or eliminate the HR increase associated with acute alcohol intoxication and that this would be independent of any effect on alcohol metabolism. METHODS: Twenty male subjects were administered placebo and alcohol (1.0 mL 95% USP ethanol/kg body weight) in a laboratory setting on one day and naltrexone (50 mg) and alcohol on another (counterbalanced). We assessed all subjects for a change in HR and for a subjective and behavioural response from 35 to 170 minutes after drug or alcohol administration. RESULTS: The placebo and alcohol mix produced a significant mean HR increase from baseline (F(1,95) = 46.01, p < 0.0001, Cohen's d = 0.62), while naltrexone and alcohol did not (nonsignificant). The significant effects of naltrexone on blood alcohol level did not account for the effect of naltrexone on alcohol-induced HR change but did account for alterations in subjective and behavioural response to alcohol. CONCLUSIONS: Naltrexone appears to substantially reduce the HR increase that is characteristic of alcohol intoxication. This finding appears to lend moderate support to the notions that, first, naltrexone has differential effects on alcohol reactions and, second, that it specifically blocks the acute psychomotor stimulant properties of alcohol. PMID- 17136217 TI - About the mechanisms of auditory verbal hallucinations: a positron emission tomographic study. AB - OBJECTIVE: Auditory verbal hallucinations (AVHs) likely result from disorders, as yet unspecified, of the neural mechanisms of language. Here we examine the functional neuroanatomy of single-word reading in patients with and without a history of AVH. METHOD: Eighteen medicated schizophrenia patients (8 with AVH and 10 without AVH) and 12 healthy control subjects were scanned with PET (15)O-water technique under 2 conditions: reading aloud English nouns and passively looking at English nouns without reading them. RESULTS: The contrast between the 2 conditions shows higher activation in Wernicke's area during the reading condition in the patient group and a reversed laterality index for the supplementary motor area in the AVH group. CONCLUSIONS: These findings provide indications about the possible mechanisms of AVH. We suggest that the abnormal laterality of the supplementary motor area activity accounts for the failure to attribute speech generated by one's own brain to one's self and that the activation of Wernicke's area accounts for the perceptual nature (hearing) of the patient's experience. PMID- 17136218 TI - The relation between peripheral and central glutamate and glutamine in healthy male volunteers. AB - OBJECTIVE: High-field strength proton magnetic resonance spectroscopy ((1)H-MRS) and peripheral blood analyses reported in the literature reveal glutamate (Glu) and glutamine (Gln) abnormalities in schizophrenia. Given the relative ease and feasibility of using peripheral measures, the present study investigates the relation between peripheral and brain Glu and Gln levels. METHODS: We recruited healthy volunteers (n = 17, mean age 21.9 [standard deviation 2.9, range 18-29] yr) between May and December 2005. All participants underwent 3 Tesla (1)H-MRS analysis with segmentation (grey matter, white matter, cerebrospinal fluid) at the Nuclear Magnetic Resonance Centre at the University of Alberta Hospital to quantify medial prefrontal cortical (mPFC) Glu and Glx (i.e., combination of Glu and Gln). Within 1 week of (1)H-MRS analysis, we collected plasma from the same participants for Glu and Gln quantification, using high-performance liquid chromatography at the Neurochemical Research Unit at the University of Alberta. RESULTS: There was no correlation between plasma Glu and either medial prefrontal cortical Glu or Glx (R(1,15) = 0.019, p = 0.944 and R(1,15) = 0.081, p = 0.757, respectively). Similarly, there was no correlation between plasma Gln and either mPFC Glu or Glx (R(1,15) = 0.029, p = 0.911 and R(1,15) = 0.025, p = 0.925, respectively). CONCLUSION: Our findings support the use of (1)H-MRS, instead of peripheral blood analysis, for investigating glutamatergic dysfunction in the brain. PMID- 17136220 TI - Concerns regarding antidepressant drug use during pregnancy. PMID- 17136222 TI - Metabolic syndrome: relevance to antidepressant treatment. PMID- 17136223 TI - Deferasirox: An effective once-daily orally active iron chelator. AB - Deferasirox (ICL670) in an orally absorbed tridentate chelator of iron (III), intended as a once-daily monotherapy for transfusional iron overload. Deferasirox was identified by Novartis from over 700 molecular entities in preclinical screening, comparing favorably with parenteral desferrioxamine or oral deferiprone. Clinical phase I and II studies demonstrated an exclusively fecal route of iron excretion, with a long plasma half-life, suitable for once-daily dosing and 24-hour protection from labile iron. Systematic large-scale prospective clinical trials have been completed in thalassemia major, sickle cell disease, and other transfusionally dependent anemias, such as myelodysplastic syndrome. These show dose-dependent reduction in body iron and have identified doses necessary either to stabilize or to decrease iron loading according to transfusion requirements. Tolerability after more than two years in phase III studies is good, with a low trial dropout rate and no drug-related arthropathy or agranulocytosis. An early, nonprogressive serum creatinine increase, remaining within normal ranges, was seen in about one-third of patients. Preliminary clinical findings using T2* as well as preclinical models suggest good drug access to myocardial iron. Deferasirox is currently registered as monotherapy for transfusional iron overload in more than 65 countries worldwide, including the United States and in the European Union. PMID- 17136224 TI - Natalizumab. AB - Medical therapeutics has entered a new and exciting era with the development of biological agents that target specific sites. In the studies performed to date, natalizumab, a humanized monoclonal antibody to alpha4 integrin, when administered intravenously at monthly intervals, appeared to be highly beneficial with respect to decreasing both relapse rate and contrast-enhancing lesions in multiple sclerosis. Salutary actions on other disease parameters were also observed. However, treatment with natalizumab was complicated by the development of a rare demyelinating disease of the brain--progressive multifocal leukoencephalopathy (PML)--that developed in three patients in clinical trials of the drug. Currently, the estimated incidence for the development of PML with natalizumab treatment is 1:1,000 after approximately 18 months of therapy; however, as the development of PML is a stochastic event, it is quite possible that higher rates will be observed after longer treatment courses. Curiously, other opportunistic infections were not observed at higher rates in the treated groups when compared to controls, suggesting a specific association between natalizumab, and perhaps other alpha4 integrin blockers, and PML. Postmarketing surveillance studies should answer this and other questions related to the use of natalizumab. PMID- 17136225 TI - Vandetanib, a novel multitargeted kinase inhibitor, in cancer therapy. AB - In clinical trials thus far, single-targeted kinase inhibitors have shown only limited success in demonstrating survival benefits in cancer. This has led to the development of multitargeted kinase inhibitors capable of disrupting various mitogenic pathways in both cancer cells and associated vasculature. Vandetanib is a novel multitargeted kinase inhibitor exhibiting potent activity against vascular endothelial growth factor receptor-2 (VEGFR-2; kinase insert domain containing receptor [KDR]) and, to a lesser extent, epidermal growth factor receptor (EGFR) and RET kinase. Vascular endothelial growth factor (VEGF) and VEGFR-2 play a pivotal role in regulating angiogenesis and vascular permeability in cancers. In addition to its antiangiogenic effects, vandetanib acts against EGFR, which is overexpressed or mutated in several solid tumors. Furthermore, vandetanib exerts activity against oncogenic RET kinase, the overexpression of which is common in medullary and papillary thyroid carcinomas. Therefore, the multitargeted kinase inhibitor vandetanib represents a new approach, targeting both tumor cells and tumor-associated endothelial cells. Preclinical studies of vandetanib have demonstrated antitumor efficacy against multiple human cancer xenografts in subcutaneous, orthotopic and metastatic models. Phase I clinical trials have demonstrated that vandetanib is well tolerated. Common adverse events included rash, diarrhea and asymptomatic QTc prolongation. Phase II clinical studies in patients with non-small-cell lung cancer have shown promising results, employing vandetanib as both monotherapy and in combination with docetaxel. Phase II studies in other cancers have likewise been initiated. This review summarizes preclinical and clinical studies of vandetanib for the treatment of cancers. PMID- 17136226 TI - Bupropion. AB - Bupropion was initially developed and licensed for the treatment of major depressive disorder in the United States in 1989. It was licensed as a pharmacotherapy for smoking cessation in the United States in 1997 and in the United Kingdom in 2000, and for the prevention of seasonal major depressive episodes in patients with seasonal affective disorder in the United States in 2006. Its main mechanism of action is believed to be via dopamine and noradrenalin reuptake inhibition. In addition to proven clinical efficacy for the treatment of major depression, the prevention of depressive episodes in patients with seasonal affective disorder, and as an aid to smoking cessation treatment, bupropion has demonstrated efficacy for attenuation of symptoms of attention deficit hyperactivity disorder, and more recently it has shown anti-inflammatory action against proinflammatory cytokines such as tumor necrosis factor-alpha (TNF alpha), which may be implicated in a number of inflammatory diseases such as Crohn's disease. The twice-daily sustained-release formulation has been extensively evaluated for smoking cessation and has shown continuous smoking abstinence rates at one year of the order of 20% across many clinical groups including healthy smokers, and smokers with cardiovascular disease, chronic obstructive airways disease, depression and schizophrenia. Bupropion is well tolerated with side effects including insomnia, headache, dry mouth, dizziness and nausea. Bupropion is a cytochrome p450 2D6 inhibitor and care must be taken when coprescribing with drugs cleared by this enzyme and when coprescribing with drugs that lower seizure threshold. Despite the clinical effectiveness and cost effectiveness of bupropion as an aid to smoking cessation, its uptake for this indication remains low when compared with nicotine replacement therapy. PMID- 17136227 TI - Role of interleukin-1 in bacterial atherogenesis. AB - The high incidence of cardiovascular diseases resulting from atherosclerosis, especially in individuals lacking classic risk factors, has spawned interest in the possibility of unrecognized risk factors, such as chronic bacterial infection. Longstanding low-grade infections, such as periodontal disease, have the potential to affect distant sites in the body by inducing host cells to release inflammatory mediators into the bloodstream. Inflammatory mediators are released by macrophages upon interaction with activated T-helper cells or upon direct recognition of bacterial antigens, and by nonimmune cells upon recognition of antigen through Toll-like receptors. One key mediator, interleukin-1 (IL-1) is released in response to bacterial infection and is known to have specific proatherogenic properties. Increased levels of IL-1 enhance vascular adhesion, vascular permeability, macrophage activation, endothelial and smooth muscle cell proliferation, and protease-induced plaque rupture - all key steps in the progression of atherogenesis. In a recent study, we demonstrated a profound reduction in the progression of atherosclerosis in IL-1 knockout mice. IL-1 holds potential as a target for future antiatherosclerotic therapies, although given its ubiquity in the body, this would not come without unwanted side effects, such as immunosuppression. PMID- 17136228 TI - A pharmacological action of vitamin E unrelated to its antioxidant capacity. AB - The pharmacological action of vitamin E on the mechanical activity of isolated guinea pig colonic smooth muscle was examined in normoxic and hypoxic conditions. In hypoxia, but not normoxia, alpha-tocopherol (1-160 microM) evoked rapid concentration-dependent contractions from the colon. This was also seen with other members of the vitamin E family, and potency measurements gave EC(50) values (microM) of 10.6 +/- 0.9 for D-alpha-tocopherol, 6.0 +/- 1.2 for D-beta tocopherol, 7.5 +/- 0.7 for D-gamma-tocopherol, and 6.1 +/- 1.5 for D-delta tocopherol. This order of potency for the components of the vitamin differs from previously studied bioassay systems and from their antioxidant activity. A range of potent natural and synthetic antioxidants was not active in this system. Compounds with structural similarities to the side chain of vitamin E produced similar stimulatory responses and some, like phytol, were more potent than the vitamin (EC(50): 1.0 +/- 0.2 microM), whereas ring structures related to the vitamin, like Trolox C, antagonized the stimulant responses in a concentration dependent manner. Therefore, this model system measures, directly, vitamin E induced responses through a mechanism that does not appear to be related to the known antioxidant capacity of these agents. PMID- 17136229 TI - Ex vivo detection of rat coronary endothelial dysfunction in diabetes mellitus- methodological considerations. AB - The present state of knowledge unequivocally indicates that chronic diabetes is associated with impaired function of coronary vessels. Langendorff retrograde perfusion is one of the most frequently employed methods to study dysfunction of coronary vasculature in animal models of diabetes mellitus. However, because of methodological discrepancies in experimental protocols, the reliability of this technique is limited. In the current study, we propose the novel technique of vasoactive drug administration and aim to evaluate its usefulness in detecting coronary dysfunction in diabetes. Using Langendorff model, we compared the results of coronary endothelium-dependent (bradykinin) and -independent (diethylamine/nitric oxide, DEA/NO) vasodilatation obtained from experimental model utilizing automatically corrected-rate infusion with commonly used, constant-rate infusion of vasoactive drug. The infusion of bradykinin at constant rate failed to reveal coronary endothelium-dependent dysfunction typical for diabetes mellitus. Induction of endothelium-independent vasodilatation by constant infusion demonstrated augmented response in diabetic hearts. The administration of bradykinin or DEA/NO at the corrected rate was associated with significantly increased maximal responses in comparison with constant infusion experiments. This phenomenon was observed particularly in the control group. We conclude that only corrected-rate infusion of vasoactive agents to actual value of coronary flow enables the reliable detection of endothelial dysfunction in diabetes mellitus. PMID- 17136230 TI - Bispecific antisense oligonucleotides with multiple binding sites for the treatment of prostate tumors and their applicability to combination therapy. AB - Antisense oligonucleotides (oligos) against transforming growth factor-alpha (TGF alpha; MR(1)) and its binding site, the epidermal growth factor receptor (EGFR; MR(2)), have proven efficacious against PC-3 and LNCaP prostate tumors when evaluated in both in vitro and in vivo models. To enhance their activity, and also to introduce a significantly different type of multifunctional agent into this field, "bispecific" oligos were constructed containing truncated sequences (derived from MR(1) and MR(2)) recognizing both TGF-alpha and EGFR mRNA internal binding sites, located about their respective AUG initiation codons. Two bispecifics were constructed, each having complementary sequences for TGF-alpha and EGFR mRNA, but differing in their 5' to 3' tandem orientation (TGF-alpha/EGFR [MR(12)] and EGFR/TGF-alpha [MR(21)] sequences). These bispecifics were tested in an in vitro system against PC-3 and LNCaP prostate tumor cells, with comparisons made to the original monospecific oligos from which they were derived. Efficacy was also compared when administered either alone or in combination with conventional chemotherapeutic agents. The purpose of this study was: 1) to validate the concept that these newly developed bispecific oligos have antitumor activity; 2) to enhance their efficacy through combination therapy; 3) to identify differences in effectiveness dependent upon binding site orientation; 4) identification of a dominant binding site that can be used to design other bispecifics that target additional tumor regulatory pathways. When fully evaluated against PC-3 cells in a series of experiments, newly developed bispecific oligos are at least as effective as their monospecific counterparts from which they were derived, and the bispecific with the MR(21) orientation is notably more effective than the MR(1) monospecific by 64% (p = 0.014 by Student t test and p = 0.068 by the more stringent Mann-Whitney U test). Bispecifics were more effective when administered with chemotherapeutics (producing inhibition of 52.1% and 61.2% for MR(12) and MR(21), respectively, with Cytoxan (cyclophosphamide) inhibition of 59.0% and 65.1% for MR(12) and MR(21), respectively, with Taxol (paclitaxel) and 63.0% and 69.4% for MR(12) and MR(21), respectively, with DES [diethylstilbestrol]). Increasing the oligo concentration above 6.25 microM with cyclophosphamide had no additional effect. The sequence directed against EGFR was dominant and contributed most to bispecific activity, particularly when inserted 5' to the TGF-alpha binding sequence (MR(21) orientation). Bispecific oligos are a significant advance in the design of antisense compounds and could play a role in treating prostate cancer, particularly when they are administered with traditional chemotherapeutics. The truncated portion of the MR(2) oligo used here should be included when constructing second-generation bispecifics that target proteins associated with other regulatory pathways, such as apoptosis. PMID- 17136231 TI - Impact of ibuprofen administration on renal drug clearance in the first weeks of life. AB - The administration of ibuprofen or any other nonselective cyclooxygenase (COX) inhibitor drug in early neonatal life is associated with a reduction of glomerular filtration, which reduces the elimination of drugs dependent on renal function for clearance. However, the relationship between COX inhibitor drug indication (prophylactic or therapeutic) and the magnitude of this effect remains unclear. Observations collected in two population pharmacokinetic studies, in preterm neonates, investigating amikacin and vancomycin were used to estimate: i) the impact of ibuprofen administration on the clearance of these drugs; and ii) the difference between prophylactic and therapeutic administration of ibuprofen on this clearance. Prophylactic administration of ibuprofen to preterm neonates on the first day of life to enhance closure of an asymptomatic patent ductus arteriosus (PDA) reduced amikacin clearance by 21% while coadministration of ibuprofen to induce closure of a symptomatic PDA resulted in an 18% reduction in vancomycin clearance in the first month of postnatal life. A significant and clinically relevant reduction in drug clearance is observed when ibuprofen is coadministered independent of indication, postmenstrual or postnatal age. Population modeling with covariate analyses can provide us with the tools to further disentangle the impact of nonselective COX-inhibitors on renal drug clearance. PMID- 17136232 TI - The effect of propofol as an antioxidant agent in intravenous regional anesthesia. AB - Intravenous regional anesthesia (IVRA) is a technique whereby a tourniquet is used to restrict blood flow to an exsanguinated limb. Propofol was shown to attenuate ischemia-reperfusion damage. We aimed to investigate the effect of low dose propofol as an antioxidant in this process. Twenty-six unpremedicated adult patients (ASA I-II) were studied. The patients in the control group (Group C, n = 12) were administered 40 ml of 0.5% lidocaine, while the patients in the propofol group (Group P, n = 14) were administered 40 ml of 0.5% lidocaine plus 20 mg propofol for IVRA. Serum levels of malondialdehyde (MDA) and paraoxonase activity were measured at 1 min before, immediately upon, and 30 min after the release of the tourniquet. Serum paraoxonase activity was observed to have a significant decreasing course in both groups (p < 0.01). In contrast, we observed a progressive increase in the serum levels of MDA in Group C (p < 0.05). However, in Group P, serum levels of MDA after the release of the tourniquet periods were significantly lower than that before the release of the tourniquet (p < 0.05). The addition of propofol (20 mg) to lidocaine for IVRA inhibits MDA levels. We conclude that the addition of propofol to lidocaine can be considered as a useful antioxidant in this type of anesthesia. PMID- 17136233 TI - Description of controlled trials published in Methods and Findings, 1979-2002. AB - It is thought that the controlled trial (CT) is the most adequate research method to assess a therapeutic intervention in terms of efficacy, and it also constitutes the basis for the development of systematic reviews on health interventions. To identify and obtain the majority of published CTs is not an easy task, mainly because of limitations concerning the currently available electronic sources. The aim of the present work was to identify, describe, and assess the quality of CTs published in the journal Methods and Findings in Experimental and Clinical Pharmacology (M&F). Additionally, to assess the retrievability of both methods, a search was performed in Medline (PubMed access) through the use of an optimal search strategy for CTs. A total of 189 original studies out of a total of 2796 reviewed articles met the CT criteria according to the Jadad scale score, we could hold that only 58% of the CTs were of good quality. The present work confirms, once again, the limitations of a CT search performed exclusively through Medline (sensitivity 64% and specificity 98%). In conclusion, we suggest that the journal M&F explicitly joins the International CONSORT Statement. PMID- 17136234 TI - Gateways to clinical trials. AB - Gateways to Clinical Trials are a guide to the most recent clinical trials in current literature and congresses. The data the following tables have been retrieved from the Clinical Trials Knowledge Area of Prous Science Integrity, the drug discovery and development portal, http://integrity.prous.com. This issues focuses on the following selection of drugs: (-)-Epigallocatechin gallate, (-) gossypol, 2-deoxyglucose, 3,4-DAP, 7-monohydroxyethylrutoside; Ad5CMV-p53, adalimumab, adefovir dipivoxil, ADH-1, alemtuzumab, aliskiren fumarate, alvocidib hydrochloride, aminolevulinic acid hydrochloride, aminolevulinic acid methyl ester, amrubicin hydrochloride, AN-152, anakinra, anecortave acetate, antiasthma herbal medicine intervention, AP-12009, AP-23573, apaziquone, aprinocarsen sodium, AR-C126532, AR-H065522, aripiprazole, armodafinil, arzoxifene hydrochloride, atazanavir sulfate, atilmotin, atomoxetine hydrochloride, atorvastatin, avanafil, azimilide hydrochloride; Bevacizumab, biphasic insulin aspart, BMS-214662, BN-83495, bortezomib, bosentan, botulinum toxin type B; Caspofungin acetate, cetuximab, chrysin, ciclesonide, clevudine, clofarabine, clopidogrel, CNF-1010, CNTO-328, CP-751871, CX-717, Cypher; Dapoxetine hydrochloride, darifenacin hydrobromide, dasatinib, deferasirox, dextofisopam, dextromethorphan/quinidine sulfate, diclofenac, dronedarone hydrochloride, drotrecogin alfa (activated), duloxetine hydrochloride, dutasteride; Edaravone, efaproxiral sodium, emtricitabine, entecavir, eplerenone, epratuzumab, erlotinib hydrochloride, escitalopram oxalate, etoricoxib, ezetimibe, ezetimibe/simvastatin; Finrozole, fipamezole hydrochloride, fondaparinux sodium, fulvestrant; Gabapentin enacarbil, gaboxadol, gefitinib, gestodene, ghrelin (human); Human insulin, human papillomavirus vaccine; Imatinib mesylate, immunoglobulin intravenous (human), indiplon, insulin detemir, insulin glargine, insulin glulisine, intranasal insulin, istradefylline, i.v. gamma-globulin, ivabradine hydrochloride, ixabepilone; LA-419, lacosamide, landiolol, lanthanum carbonate, lidocaine/prilocaine, liposomal cisplatin, lutropin alfa; Matuzumab, MBP(82-98), mecasermin, MGCD-0103, MMR-V, morphine hydrochloride, mycophenolic acid sodium salt; Natalizumab, NCX-4016, neridronic acid, nesiritide, nilotinib, NSC-330507; O6-benzylguanine, olanzapine/fluoxetine hydrochloride, omalizumab; Panitumumab, parathyroid hormone (human recombinant), parecoxib sodium, PEG filgrastim, peginterferon alfa-2a, peginterferon alfa-2b, pegvisomant, pemetrexed disodium, perospirone hydrochloride, pexelizumab, phorbol 12-myristate 13 acetate, pneumococcal 7-valent conjugate vaccine, posaconazole, pramiconazole, prasugrel, pregabalin, prilocaine; rAAV-GAD65, raclopride, rasagiline mesilate, retapamulin, rosuvastatin calcium, rotigotine, rufinamide; SarCNU, SB-743921, SHL 749, sirolimus-eluting stent, sitaxsentan sodium, sorafenib; TachoSil, tadalafil, talampanel, Taxus, tegaserod maleate, telithromycin, telmisartan/hydrochlorothiazide, temsirolimus, tenatoprazole, teriflunomide, tetrathiomolybdate, ticilimumab, timcodar dimesilate, tipifarnib, tirapazamine, TPI, tramiprosate, trifluridine/TPI, trimethoprim; Ularitide, Urocortin 2; Valdecoxib, valganciclovir hydrochloride, valproate magnesium, valspodar, vardenafil hydrochloride hydrate, vitespen, vofopitant hydrochloride, volociximab, vorinostat; Yttrium 90 (90Y) ibritumomab tiuxetan; Ziprasidone hydrochloride, zotarolimus, zotarolimus-eluting stent. PMID- 17136235 TI - Noninjectable methods of insulin administration. AB - Optimal glycemic control is essential for the prevention of diabetes-related complications, and the intensive insulin regimens that best resemble physiological insulin secretion are most likely to attain it. However, there are many limitations that preclude the early use of insulin by patients with type 2 diabetes or wider implementation of the intensive regimens in type 1 diabetes. More acceptable alternative routes of insulin administration and effective, noninvasive, patient-friendly delivery systems that alleviate the burden of insulin injections have been researched over the years. To date, only pulmonary inhalation of insulin has become a feasible alternative; it has proved to be as effective and well tolerated as the subcutaneously injected regular insulin and it has a pharmacodynamic profile well suited for mealtime insulin therapy. Several pulmonary insulin delivery systems are in different stages of development, and one (Exubera, Nektar Therapeutics/Pfizer Inc.-Sanofi-Aventis Group) has already become clinically available in the United States and Europe for patients with diabetes. Other noninjectable methods of insulin administration are reviewed. PMID- 17136236 TI - Bleeding in acute coronary syndromes. PMID- 17136237 TI - Synthesis and stereoselective functionalization of silylated heterocycles as a new class of formyl anion equivalents. AB - The fluoride ion-induced reactivity of a series of silyl heterocycles leads to the generation of nucleophilic species capable of interacting with electrophiles, thus disclosing new classes of formyl and acyl anion synthons. Moreover, when reacting stereodefined molecules, the stereoinformation of the reacting carbon silicon bond is transferred to the newly formed carbon-carbon bond, suggesting possible applications in stereoselective synthesis. Thus, silyl dithiolanes, oxathiolanes, dioxolanes, thiazolidines and oxazolidines can be efficiently and stereoselectively functionalized under fluoride ion conditions in the presence of electrophiles. While direct access to silyl heterocycles is generally either prevented or troublesome, a novel protocol for their synthesis has also been developed, together with a simple general access route to several functionalized and stereodefined mercaptans, building blocks for the construction of silyl heterocycles. PMID- 17136238 TI - [Cu(HF2)(pyz)2]BF4 (pyz = pyrazine): long-range magnetic ordering in a pseudo cubic coordination polymer comprised of bridging HF2- and pyrazine ligands. AB - [Cu(HF2)(pyz)2]BF4 consists of rare mu(1,3) bridging HF2- anions and micro pyrazine ligands leading to a 3D pseudo-cubic framework that antiferromagnetically orders below 1.54(1) K. PMID- 17136239 TI - Enzymatically triggered self-assembly of poly(ethylene glycol)-attached oligopeptides into well-organized nanofibers. AB - A unique and programmable peptide self-assembling system has been fabricated by using poly(ethylene glycol)-attached amphiphilic oligopeptide, which shows rapid self-assembly into well-organized beta-sheet nanofibers in response to an enzymatic reaction. PMID- 17136240 TI - Carborane functionalized pyrroles and porphyrins via the Suzuki cross-coupling reaction. AB - A new route to carboranylated pyrroles and porphyrins is reported which involves the Suzuki coupling of readily available bromo- and boronic acid-pyrroles and bromoporphyrins with functionalized carboranes; the X-ray structures of two targeted products are presented and discussed. PMID- 17136241 TI - A supramolecular hydrogel that responds to biologically relevant stimuli. AB - First demonstration of heat and pH-responsive hydrogel of SDS and a zwitterionic amphiphile, sodium N-(n-dodecyl-2-aminoethanoyl)-L-valinate with very low minimum gelation concentration. PMID- 17136242 TI - A novel 3D porous metal-organic framework based on trinuclear cadmium clusters as a promising luminescent material exhibiting tunable emissions between UV and visible wavelengths. AB - A novel 3D porous MOF built with trinuclear cadmium clusters exhibiting rhombohedral topology has been synthesized and characterized as a promising luminescent material that can give tunable emissions between UV and visible wavelengths by controlling the number of guest molecules. PMID- 17136243 TI - An enantioselective desymmetrisation approach to C9-substituted trans-hydrindene rings based on a diastereotopic group-selective intramolecular Diels-Alder reaction. AB - The synthesis of an achiral skipped bis(1,3-diene) substrate was achieved, which was shown to undergo an enantioselective, diastereotopic group-selective, intramolecular Diels-Alder reaction. PMID- 17136244 TI - Supramolecular metallomesogens. Self-organization of an H-bonded tetrameric assembly into columnar phase from single component. AB - The first example of supramolecular metallomesogens derived from palladium complexes exhibiting columnar phase is reported, and the formation of the mesophase is induced by H-bonding in a tetrameric assembly. PMID- 17136245 TI - Synthesis and structural characterisation as 12-helix of the hexamer of a beta amino acid tethered to a pyrrolidin-2-one ring. AB - Starting from (3S,4R,1'S)-3-amino-2-oxo-1-[1'-(4-methoxyphenylethyl)]pyrrolidine carboxylic acid (2), the first synthesis of a beta-foldamer containing pyrrolidin 2-one rings is described, whose 12-helix conformation is assigned by NMR analysis and confirmed by molecular dynamics (MD) simulations. PMID- 17136246 TI - Polymorphs of 1,1-bis(4-hydroxyphenyl)cyclohexane and multiple Z' crystal structures by melt and sublimation crystallization. AB - Close packing conflict in a metastable polymorph of the pure title host (Z' = 2, melt crystal m) is resolved in the stable form (Z' = 1, sublimed crystal s) as O H...O hydrogen bond changes to O-H...pi interaction. Melt crystallization and sublimation show a greater percentage of high Z' structures in CSD statistics. PMID- 17136247 TI - Racemic progesterone: predicted in silico and produced in the solid state. AB - A computational prediction that mixing the synthetic mirror image of progesterone with its natural form would produce a specific racemic crystal structure was validated. PMID- 17136248 TI - A convenient enzymatic synthesis of L-halotryptophans. AB - A scalable and general biotransformation for the generation of a series of L halotryptophans using the lysate of a commercially available microorganism containing tryptophan synthase. PMID- 17136249 TI - Acidic iridium hydrides: implications for aerobic and Oppenauer oxidation of alcohols. AB - [Cp*Ir(H)(bpym)]+ and [Cp*Ir(H)(bpy)]+ are the first examples of iridium based catalysts for the aerobic oxidation of alcohols; the catalytic cycle proceeds via acidic hydrides. Deprotonation of the hydride leads to a highly oxygen sensitive Ir I species that regenerate the Ir III complexes upon oxidation with dioxygen. PMID- 17136250 TI - A simple asymmetric organocatalytic approach to optically active cyclohexenones. AB - Optically active 2,5-disubstituted-cyclohexen-2-one derivatives have been prepared in a one-pot process consisting of five reaction steps: an organocatalytic asymmetric conjugated addition of beta-ketoesters to alpha,beta unsaturated aldehydes that proceeds in aqueous solutions or under solvent-free conditions has been implemented in a multi-step process. PMID- 17136251 TI - A chirality-induced alpha phase and a novel molecular magnetic metal in the BEDT TTF/tris(croconate)ferrate(III) hybrid molecular system. AB - The novel paramagnetic and chiral anion [Fe(C5O5)3]3- has been combined with the organic donor BEDT-TTF (= ET = bis(ethylenedithio)tetrathiafulvalene) to yield the first chirality-induced alpha phase and a paramagnetic metal. PMID- 17136252 TI - Dioxygenase-catalysed dihydroxylation of arene cis-dihydrodiols and acetonide derivatives: a new approach to the synthesis of enantiopure tetraoxygenated bioproducts from arenes. AB - cis-Dihydrodiols of anthracene and benz[a]anthracene, and acetonide derivatives of the cis-dihydrodiols of benzene, fluorobenzene, biphenyl and phenanthrene have been identified as substrates for dioxygenase enzymes, yielding the corresponding enantiopure arene bioproducts, bis(cis-dihydrodiol)s and cis-diol acetonides respectively. PMID- 17136253 TI - Dispersing as-prepared single-walled carbon nanotube powders with linear conjugated polymers. AB - Suitably modified linear conjugated poly(arylene ethynylene)s are able to assist effective debundling and dispersion of crude as-prepared single-walled carbon nanotube powders in organic solvents, the dispersion of which is effected via a surface coating mechanism and, to some extent, in a size-selective fashion. PMID- 17136254 TI - Elucidation of the extraordinary 4-membered pyrrole ring-contracted azeteoporphyrinoid as an intermediate in chlorin oxidation. AB - Reaction of 2,3-dioxochlorins with benzeneselenic anhydride (BSA) results in the formation of unusual ring-contracted azetine derivatives that further react with BSA to afford porpholactones. PMID- 17136255 TI - Preparation and photoactivity of nanostructured anatase, rutile and brookite TiO2 thin films. AB - Photoactive films consisting of pure anatase, brookite or rutile TiO2 were prepared by dip coating from water dispersions obtained by using TiCl4 as the precursor under similar mild experimental conditions. PMID- 17136256 TI - An unusually high thermal stability within a novel lanthanide 1,3,5 cyclohexanetricarboxylate framework: synthesis, structure, and thermal data. AB - A novel three-dimensional lanthanide-organic framework has been synthesized; this material has an exceptionally high thermal stability (600 degrees C) and an unusually low coordination number for a lanthanide ion (CN = 6). PMID- 17136257 TI - Modification of multi-wall carbon nanotube surfaces with poly(amidoamine) dendrons: synthesis and metal templating. AB - Hydroxyl functional poly(amidoamine) dendron wedges have been reacted with the surface of multi-walled carbon nanotubes and used for templating silver nanoparticles. PMID- 17136258 TI - Iodine catalyzed one-pot diamination of glycals with chloramine-T: a new approach to 2-amino-beta-glycosylamines for applications in N-glycopeptide synthesis. AB - Iodine catalyzes a facile one-pot direct diamination of glycals with chloramine-T to afford stereoselectively 2-amino-beta-glycosylamine derivatives that serve as convenient precursors for the synthesis of N-linked glycopeptides. PMID- 17136259 TI - Selective oxidation of benzene to phenol over FeAlPO catalysts using nitrous oxide as oxidant. AB - A tetrahedrally coordinated iron in framework substituted microporous AlPO-5 catalysts are shown to be active and selective for the hydroxylation of benzene to phenol, using nitrous oxide as the oxidant. PMID- 17136260 TI - Enhanced catalase-like activity of manganese salen complexes in water: effect of a three-dimensionally fixed auxiliary. AB - A new Mn(Salen) complex bearing an ureido group as an auxiliary that is three dimensionally fixed by a cyclopentane ring fused to the Salen structure was developed. This compound exhibited considerably higher catalase-like activity than the original Mn(Salen), i.e., the cyclopentane-fused Mn(Salen) without the auxiliary, under near-physiological conditions. PMID- 17136261 TI - Guest-dependent inversion rate of a tetranuclear single metallohelicate. AB - Complexation of linear hexaoxime ligand H6L with Zn2+ and Mn+ (= La3+, Ba2+) afforded a tetranuclear single metallohelicate [LZn3M]n+, whose inversion rate can be modulated by the central metal Mn+. PMID- 17136262 TI - An unexpected synthesis of ketene monothioacetals. AB - Some dithiocarbonates (xanthates) can be converted into ketene monothioacetals through extrusion of sulfur upon treatment with base and an alkylating agent. PMID- 17136263 TI - A self-assembled hexadecanuclear 4 x[2 x 2] Mn(II)16 square grid from a pyridazine bis(hydrazone) ligand: synthesis, structure and magnetism. AB - A compartmentalized {4 x[2 x 2]} Mn(II)16 antiferromagnetically coupled square grid results from the self-assembly reaction of a tetratopic pyridazine bis(hydrazone) ligand and Mn(II). PMID- 17136264 TI - Gold catalysts for pure hydrogen production in the water-gas shift reaction: activity, structure and reaction mechanism. AB - The production of hydrogen containing very low levels of carbon monoxide for use in polymer electrolyte fuel cells requires the development of catalysts that show very high activity at low temperatures where the equilibrium for the removal of carbon monoxide using the water-gas shift reaction is favourable. It has been claimed that oxide-supported gold catalysts have the required high activity but there is considerable uncertainty in the literature about the feasibility of using these catalysts under real conditions. By comparing the activity of gold catalysts with that of platinum catalysts it is shown that well-prepared gold catalysts are significantly more active than the corresponding platinum catalysts. However, the method of preparation and pre-treatment of the gold catalysts is critical and activity variations of several orders of magnitude can be observed depending on the methods chosen. It is shown that an intimate contact between gold and the oxide support is important and any preparative procedure that does not generate such an interaction, or any subsequent treatment that can destroy such an interaction, may result in catalysts with low activity. The oxidation state and structure of active gold catalysts for the water-gas shift reaction is shown to comprise gold primarily in a zerovalent metallic state but in intimate contact with the support. This close contact between small metallic gold particles and the support may result in the "atoms" at the point of contact having a net charge (most probably cationic) but the high activity is associated with the presence of metallic gold. Both in situ XPS and XANES appear unequivocal on this point and this conclusion is consistent with similar measurements on gold catalysts even when used for CO oxidation. In situ EXAFS measurements under water gas shift conditions show that the active form of gold is a small gold cluster in intimate contact with the oxide support. The importance of the gold/oxide interface is indicated but the possible role of special sites (e.g., edge sites) on the gold clusters cannot be excluded. These may be important for CO oxidation but the fact that water has to be activated in the water gas shift reaction may point towards a more dominant role for the interfacial sites. The mechanism of the water gas shift reaction on gold and other low temperature catalysts has been widely investigated but little agreement exists. However, it is shown that a single "universal" model is consistent with much of the experimental literature. In this, it is proposed that the dominant surface intermediate is a function of reaction conditions. For example, as the temperature is increased the dominant species changes from a carbonate or carboxylate species, to a formate species and eventually at high temperatures to a mechanism that is characteristic of a redox process. Similar changes in the dominant intermediate are observed with changes in the gas composition. Overall, it is shown that reported variations in the kinetics, structure and reaction mechanism for the water gas shift reaction on gold catalysts can now be understood and rationalised. PMID- 17136265 TI - Short-range interactions between non-ionic surfactant layers. AB - Short-range interactions between surfactant and lipid layers are of great importance in technical applications in complex fluids such as foams, dispersions and emulsions, as well as in the formulation and performance of dispersants, detergents and flocculants. It is also of utmost importance in biological systems where interactions between biomembranes influence a range of processes. The field of short-range interactions has been thoroughly investigated during the past 30 years, following the emergence of a number of techniques to measure interaction forces. Thus, our understanding has increased considerably and it is timely to summarize relevant knowledge accumulated in this area. In this review we focus on the nature of short-range interactions between non-ionic and zwitterionic surfactant and lipid layers exposing their polar groups to the surrounding medium. We discuss the complex interplay of short-range (van der Waals, hydration, steric and other) forces based on recent theoretical and experimental results. PMID- 17136266 TI - Computational assessment of the entropy of solvation of small-sized hydrophobic entities. AB - A high level polarizable force field is used to study the temperature dependence of hydrophobic hydration of small-sized molecules from computer simulations. Molecular dynamics (MD) simulations of liquid water at various temperatures form the basis of free energy perturbation calculations that consider the onset and growth of a repulsive sphere. This repulsive sphere acts as a model construct for the hydrophobic species. In the present study, an extension is pursued for seven independent target temperatures, ranging from close to the freezing point almost up to the boiling point of liquid water under standard conditions. Care is taken to maintain proper physico-chemical model descriptions by cross-checking with experimental water densities at the selected target temperatures. The polarizable force field description of molecular water turns out to be suitable throughout the entire temperature domain considered. Derivatives of the computed free energies of hydrophobic hydration with respect to the temperature give access to the changes in entropy. In practice the entropy differential is determined from the negative of the slope of tangential lines formed at a certain target temperature in the free energy profile. The obtained changes in entropy are negative for small-sized cavities, and hence reconfirm the basic ideas of the Lum Chandler-Weeks theory on hydrophobic hydration of small-sized solutes. PMID- 17136267 TI - Theoretical modeling of energy redistribution and stereodynamics in CF scattering from Si(100) under grazing incidence. AB - We have simulated CF scattering from Si(100) using the molecular dynamics method. Translational energy loss spectra are presented. The shape of the energy loss distribution as a result of internal energy release is analyzed. At the classical turning point, the internal energy of the molecule is mainly in the form of rotational energy. The strong rotational excitation results in additional molecule-surfaces interactions during the latter half of the collision. These additional collisions permit some molecules that initially gain internal energy exceeding the bond strength to ultimately survive the collision process via rotational de-excitation. The rotational motion exhibited by surviving molecules is determined by the combination of the molecular axis orientation and the local surface structure during the collision process. The rotation planes of the surviving molecules are preferentially aligned with the surface normal (cartwheel like and propeller-like motions). In this study, propeller-like motion of the surviving molecules is predicted. The majority of surviving molecules exhibit a cartwheel-like motion. However, molecules that gain a propeller-like rotation exhibit a much better alignment of their planes-of-rotation compared with molecules exhibiting cartwheel-like motion. PMID- 17136268 TI - On the site-specificity of polycarbonyl complexes in Cu/zeolites: combined experimental and DFT study. AB - The preferred Cu(+) sites and formation of mono-, di-, and tricarbonyl complexes in the Cu-FER were investigated at the periodic density functional theory level and by means of FTIR spectroscopy. The site-specificity of adsorption enthalpies of CO on Cu-FER and of vibrational frequencies of polycarbonyl complexes were investigated for various Cu(+) sites in Cu-FER. Large changes in the Cu(+) interaction with the zeolite framework were observed upon the formation of carbonyl complexes. The dicarbonyl complexes formed on Cu(+) in the main channel or on the intersection of the main and perpendicular channels are stable and both, adsorption enthalpies and CO stretching frequencies are not site-specific. The fraction of Cu(+) ions in the FER cage, that cannot form dicarbonyl can be determined from IR spectra (about 7% for the Cu-FER with Si/Al = 27.5 investigated here). The tricarbonyl complexes can be formed at the Cu(+) ions located at the 8-member ring window at the intersection of main and perpendicular channel. The stability of tricarbonyl complexes is very low (DeltaH degrees (0 K)>or=-4 kJ mol(-1)). PMID- 17136269 TI - Conformational heterogeneity and low-frequency vibrational modes of proteins. AB - Molecular dynamics simulation and normal mode analysis are used to calculate the vibrational density of states of dihydrofolate reductase complexed with nicotinamide adenine dinucleotide phosphate at 120 K and the results are compared with the experimental spectrum derived from inelastic neutron scattering. The simulation results indicate that the experimental spectrum arises from an average over proteins trapped in different conformations with structural differences mainly in the loop regions, and that these conformations have significantly different low-frequency (<20 cm(-1)) spectra. Thus, the experimentally measured spectrum is an average over the vibrational modes of different protein conformations and is thus inhomogeneously broadened. The implications of this broadening for future neutron scattering experiments and ligand binding calculations are discussed. PMID- 17136270 TI - The photodissociation dynamics of O2 at 193 nm: an O3PJ angular momentum polarization study. AB - In the following paper we present translational anisotropy and angular momentum polarization data for O((3)P(1)) and O((3)P(2)) products of the photodissociation of molecular oxygen at 193 nm. The data were obtained using polarized laser photodissociation coupled with resonantly enhanced multiphoton ionization and velocity-map ion imaging. Under the jet-cooled conditions employed, absorption is believed to be dominated by excitation into the Herzberg continuum. The experimental data are compared with previous experiments and theoretical calculations at this and other wavelengths. Semi-classical calculations performed by Groenenboom and van Vroonhoven [J. Chem. Phys, 2002, 116, 1965] are used to estimate the alignment parameters arising from incoherent excitation and dissociation and these are shown to agree qualitatively well with the available experimental data. Following the work of Alexander et al. [J. Chem. Phys, 2003, 118, 10566], orientation and alignment parameters arising from coherent excitation and dissociation are modelled more approximately by estimating phase differences generated subsequent to dissociation via competing adiabatic pathways leading to the same asymptotic products. These calculations lend support to the view that large values of the coherent alignment moments, but small values of the corresponding orientation moments, could arise from coherent excitation of (and subsequent dissociation via) parallel and perpendicular components of the Herzberg I, II and III transitions. PMID- 17136271 TI - Reactivity induced at 25 K by low-energy electron irradiation of condensed NH3 CH3COOD (1 : 1) mixture. AB - Chemical reactivity is observed following electron irradiation of a binary mixture of ammonia (NH(3)) and acetic acid (CH(3)COOD) at 25 K, without any subsequent thermal activation, as evidenced by vibrational high resolution electron energy loss spectroscopy (HREELS). Analysis of the HREEL spectra and comparison with infrared and Raman data of different molecules are compatible with glycine formation in its zwitterionic form. The onset for electron induced reaction is found to be at about approximately 13 eV. The mechanisms may involve NH radicals interaction with CH(3)COOD molecules. Then glycine formation does not imply any displacement of reactants, so that it involves only NH(3) and CH(3)COOD neighboring molecules. PMID- 17136272 TI - Photoluminescence spectra of a conjugated polymer: from films and solutions to single molecules. AB - The purpose of this work is to address the issue of applicability of single molecule spectroscopy (SMS) results for conjugated polymers to "bulk" samples, e.g. conjugated polymer films. Also, some apparent inconsistencies in the literature on SMS regarding the photoluminescence spectral position of conjugated polymers are discussed. We present a series of photoluminescence spectra of thin films of the conjugated polymer poly(2-methoxy-5-(2'-ethyl-hexyloxy)-1,4 phenylene-vinylene) (MEH-PPV) with a wide range of varying thickness. The thickness was varied from approximately 20 nm to the value corresponding to well separated single molecules (SMS sample). The thickness variation resulted in a strong ( approximately 2000 cm(-1)) blue-shift and broadening of the spectrum. The result was reproduced on isolated molecules embedded into a PMMA matrix. This effect cannot be explained by a decrease in energy transfer "freedom" alone. We performed a comprehensive comparison of presented and elsewhere published spectra of MEH-PPV polymer and oligomers in different samples: films, solutions, isolated molecule coatings and standard SMS samples. The comparison allows that the main reason behind the blue shift is conformational disorder, which is largely dependent on the sample. We also discuss some experimental aspects of SMS, such as representativeness of detected molecules, spectral sensitivity of a setup and temperature. Together with differences in sample preparation method, these issues can explain the existing inconsistencies in the literature. PMID- 17136273 TI - Introducing the Time-resolved methods in biophysics series. PMID- 17136274 TI - Time-resolved methods in Biophysics. 1. A novel pump and probe surface-enhanced resonance Raman approach for studying biological photoreceptors. AB - This article describes a method, based on surface-enhanced resonance Raman (SERR) spectroscopy, for studying the reaction dynamics of photoreceptors immobilized on metal electrodes. Time resolution and fresh sample conditions are achieved by synchronizing the rotational and translational motion of a novel kinematic electrode with the duration and time delay between the pump and probe events. The power and sensitivity of the method is illustrated by studying the photocycle of the sensory photoreceptor NpSRII and its sensitivity to the applied electric field. The results are compared with time-resolved resonance Raman measurements in solution. PMID- 17136275 TI - Time-resolved methods in Biophysics. 2. Monitoring haem proteins at work with nanosecond laser flash photolysis. AB - Haem proteins have long been the most studied proteins in biophysics, and have become paradigms for the characterization of fundamental biomolecular processes as ligand binding and regulatory conformational transitions. The presence of the haem prosthetic group, the absorbance spectrum of which has a ligation sensitive region conveniently located in the UV-visible range, has offered a powerful and sensitive tool for the investigation of molecular functions. The central Fe atom is capable of reversibly binding diatomic ligands, including O(2), CO, and NO. The Fe-ligand bond is photolabile, and a reactive unligated state can be transiently generated with a pulsed laser. The photodissociated ligands quickly rebind to the haem and the process can be monitored by transient absorbance methods. The ligand rebinding kinetics reflects protein dynamics and ligand migration within the protein inner cavities. The characterization of these processes was done in the past mainly by low temperature experiments. The use of silica gels to trap proteins allows the characterization of internal ligand dynamics at room temperature. In order to show the potential of the laser flash photolysis techniques, combined with modern numerical analysis methods, we report experiments conducted on two non-symbiotic haemoglobins from Arabidopsis thaliana. The comparison between time courses recorded on haemoglobins in solution and encapsulated in silica gels allows for the highlighting of different interplays between protein dynamics and ligand migration. PMID- 17136276 TI - Foreword to the fullerene special issue. PMID- 17136277 TI - Photoinduced charge and energy transfer involving fullerene derivatives. AB - In this feature article, a brief overview over the photoinduced energy and charge transfer mechanisms involving fullerenes will be presented. The photoinduced charge separation between organic donor and acceptor molecules is the basic photophysical mechanism for natural photosynthesis and nearly all organic solar cell concepts. We will give a short introduction to the mechanisms of excited state charge transfer and resonant energy transfer and present examples of relevant applications in organic optoelectronics and photodynamic tumor therapy. PMID- 17136278 TI - Voltammetric characterization of C60(PhX)2 (X = H, Br) and digital simulation of their electrochemically-induced reactivity. AB - The electrochemistry of two new 1,7-diaryl C(60) phenylated derivatives is explored in THF at various temperatures (from 25 to -90 degrees C). While at room temperature their voltammetric response is that typically shown by fairly stable species, when the temperature drops a very high electrochemically induced reactivity is evidenced. The investigation of the voltammetric patterns supported by an extensive use of digital simulation techniques finally led to the formulation of a reaction mechanism that includes electrochemically-induced migration of the phenyl groups as a possible explanation of the observed behavior. PMID- 17136279 TI - A dendritic fullerene-porphyrin dyad. AB - We describe the synthesis, characterization and photophysical properties of a fullerene derivative whose structure includes a Zn-porphyrin and a second generation liquid-crystalline (LC) dendrimer. The size of the fullerene and porphyrin units with respect to the size of the LC dendrimer prevents the formation of liquid-crystalline phases. However, this system gives interesting photoinduced electron transfer phenomena. Compound has been investigated by steady state and time resolved fluorescence as well as transient absorption spectroscopy in polar and apolar solvents. We demonstrate that the fluorescence of the porphyrin unit in is quenched compared to the Zn-tetraphenylporphyrin used as reference. Femto- and picosecond transient absorption permit to identify the formation of a radical ion pair while nanosecond experiments allowed the determination of the charge recombination lifetimes. PMID- 17136280 TI - Charge separation and energy transfer in a caroteno-C60 dyad: photoinduced electron transfer from the carotenoid excited states. AB - We have designed and synthesized a molecular dyad comprising a carotenoid pigment linked to a fullerene derivative (C-C(60)) in which the carotenoid acts both as an antenna for the fullerene and as an electron transfer partner. Ultrafast transient absorption spectroscopy was carried out on the dyad in order to investigate energy transfer and charge separation pathways and efficiencies upon excitation of the carotenoid moiety. When the dyad is dissolved in hexane energy transfer from the carotenoid S(2) state to the fullerene takes place on an ultrafast (sub 100 fs) timescale and no intramolecular electron transfer was detected. When the dyad is dissolved in toluene, the excited carotenoid decays from its excited states both by transferring energy to the fullerene and by forming a charge-separated C.+ -C(60).- . The charge-separated state is also formed from the excited fullerene following energy transfer from the carotenoid. These pathways lead to charge separation on the subpicosecond time scale (possibly from the S(2) state and the vibrationally excited S(1) state of the carotenoid), on the ps time scale (5.5 ps) from the relaxed S(1) state of the carotenoid, and from the excited state of C(60) in 23.5 ps. The charge-separated state lives for 1.3 ns and recombines to populate both the low-lying carotenoid triplet state and the dyad ground state. PMID- 17136281 TI - Metal ion dependent fluorescence quenching in a crown ether bridged porphyrin fullerene dyad. AB - The fluorescence decay kinetics from a benzonitrile solution of a dibenzo-18 crown-6 ether bridged porphyrin-fullerene dyad has been studied in the presence of a range of metal ions. Dual-exponential fluorescence decay behaviour has been attributed to conformational flexibility of the molecule influencing quenching interactions between the photo-excited porphyrin and fullerene. Additions of sodium, potassium and lithium ions significantly modulate the observed fluorescence decay processes while the larger tetrabutylammonium ion has only a minor affect. The results are discussed in terms of ion inclusion within the crown ether affecting both the bridge conformational properties and donor acceptor electronic interactions. PMID- 17136282 TI - Rhenium(I) and ruthenium(II) complexes with a crown-linked methanofullerene ligand: synthesis, electrochemistry and photophysical characterization. AB - A cyclopropanation reaction has been used to prepare two methanofullerenes bearing a 2,2'-bipyridine () or pyridine () ligand separated from the fullerene through an oxyethylene macrocyclic spacer. Derivatives and were, in turn, employed to synthesize two fullerene-based ruthenium(ii) and rhenium(i) donor acceptor dyads whose molecular structure was confirmed by (1)H NMR, (13)C NMR and exact mass determination. The UV-Vis spectrum of the dyads is the superimposition of those of appropriate model systems, indicating that ground-state electronic interactions between the constituent chromophores, in solution, are negligible, in line also with the electrochemical results. The complex voltammetric pattern was characterized by the superimposition of signals attributed to one moiety or another without significant shifts with respect to their models. Furthermore, both species undergo partial chemical degradation in the time scale of cyclic voltammetry upon their multiple reduction. Photophysical properties of and , namely, excited state interactions between the ruthenium(ii) or rhenium(i) complexes and [60]fullerene have been investigated by steady-state and time resolved UV-Vis-NIR luminescence spectroscopy that was complemented by nanosecond laser flash photolysis in CH(2)Cl(2) solutions. All experimental findings were set into relation with the corresponding reference compounds. More precisely, excitation of the metal complexes in and gives rise to a notable steady-state and time-resolved luminescence quenching of both metal to ligand charge transfer states (i.e., [Ru(bpy)(3)](2+) and [(bpy)Re(CO)(3)(py)](+)). Conclusive evidence about the nature of the photoproducts came from nanosecond laser flash photolysis. In these experiments, only the long-lived and oxygen-sensitive [60]fullerene triplets were detected. Two pathways are envisioned for this [60]fullerene triplet formation. Firstly, intramolecular transduction of the triplet excited state energy evolving from the photoexcited metal complexes. Secondly, intersystem crossing of directly excited [60]fullerene. PMID- 17136283 TI - Photoinduced energy and electron transfer in fullerene-oligophenyleneethynylene systems: dependence on the substituents of the oligomer unit. AB - The photophysical properties of fullerene hybrid systems in which disymmetrically substituted linear oligophenyleneethynylene (OPE) substituents have been attached to C(60) through a pyrrolidine ring are discussed. These hybrid systems differ in both the length of the conjugated OPE backbone and in the type of terminating groups employed, i.e. tri-isopropylsilane (-Si(iPr)(3)) and N,N-di-n-butylaniline (PhN(nBu)(2)). The terminating group is found to be crucial in determining the fate of light absorbed by the hybrid. In CH(2)Cl(2) and benzonitrile, the PhN(nBu)(2) terminated hybrids undergo electron transfer with charge separation lasting as long as 390 ns in the more polar medium, as detected via near-infrared transient absorption spectroscopy. Under the same conditions the Si(iPr)(3) terminated hybrids show ultrafast OPE --> C(60) singlet energy transfer (k = 10(9)-10(10) s(-1)) followed by regular deactivation of the C(60) moiety, as determined via UV-VIR-NIR steady state and time-resolved spectroscopy. Only in polar benzonitrile such systems can undergo electron transfer to some extent (40% yield). The results here presented can be readily explained in light of the electrochemical properties of the hybrids. The low oxidation potentials of the PhN(nBu)(2) terminated systems allow the formation of low lying charge separated states ( approximately 1.45 eV) which, in Si(iPr)(3) terminated analogues, are shifted substantially upward ( approximately 1.90 eV) and become hardly accessible via direct excitation or sensitization of the C(60) singlet level (1.72 eV). These results, when examined in light of the performance of photovoltaic devices using these hybrids as active materials, show a nice structure-activity relationship supporting the appeal of the so-called molecular approach to photovoltaic devices. PMID- 17136284 TI - Photophysical and electrochemical properties of a fullerene-stoppered rotaxane. AB - The photophysical and electrochemical properties of a fumaramide rotaxane stoppered with C(60) are reported. The results evidenced the strong binding interactions between the template and the macrocycle, which are also supported by molecular modelling. PMID- 17136285 TI - Optical spectrum of C60 mono-adducts: assignment of transition bands using time resolved EPR magneto-photo-selection. AB - The magneto-photo-selection technique implemented in the time-resolved EPR (TR EPR) experiment is used for studying the characteristics of the optical spectrum of C(60) mono-adducts, in the 410-690 nm wavelength range. The analysis of the shape of the triplet state TR-EPR spectra of the mono-adducts, recorded after laser light pulses having polarization parallel or perpendicular to the magnetic field direction allows to determine the orientation distribution of the excited molecules; whence the direction of transition moments in the molecular frame is inferred. This information provides the assignment of the vibronic states symmetries. PMID- 17136286 TI - Ionisation of fullerenes and fullerene clusters using ultrashort laser pulses. AB - We give a brief review of the literature concerning the ultra-short pulse ionisation of fullerenes in the gas phase. Emphasis is placed on the excitation time dependence of different ionisation regimes as manifested by photoelectron spectroscopy. The ionisation rates are modelled for the intermediate situation where the excitation energy is equilibrated between electronic degrees of freedom but not yet coupled to vibrational degrees of freedom. The model is shown to describe many aspects of the experiments. New results are presented on the intra cluster molecular fusion of fullerene molecules when van der Waals bound clusters of fullerenes are exposed to ultra-short laser pulses. Pump-probe measurements give a decay time constant for the intra-cluster fusion reaction of 520 +/- 55 fs. A comparison with monomer ionisation results suggests that the time window for the fusion reaction is influenced by the coupling of the electronic excitation energy to vibrational degrees of freedom of the molecules in the cluster. PMID- 17136288 TI - Anti-IgE monoclonal antibody for the treatment of the asthma and other manifestations related to allergic diseases. AB - OBJECTIVES: To report on the pharmacology, efficacy and safety of omalizumab, a new option for the treatment of asthma and allergic diseases and the first monoclonal anti-IgE antibody approved for clinical use. SOURCES: MEDLINE, a non systematic search including reviews and original papers, chosen according to their relevance in the authors' opinion. SUMMARY OF THE FINDINGS: The paper emphasizes the central role IgE plays in allergic diseases and the biological rationale for its use, the evidence upon which the current recommendations for the use of anti-IgE in uncontrolled asthma are based and its possible future applications, in addition to the recommendation that in clinical practice doses must be adjusted for weight and serum IgE levels. Omalizumab was approved in Brazil for patients with severe uncontrolled asthma presenting with a positive skin prick test for one or more relevant aeroallergen, or IgE specific to a relevant allergen detected in serum, having a total IgE level of between 30 and 700 UI/mL. For the time being its use should be restricted to patients aged 12 years or more, but there are prospects that it will be licensed for use with children over 6 years old. CONCLUSIONS: Some severe asthma cases cannot be controlled with the regular treatment options aimed at preventing symptoms and exacerbations, and so require frequent or prolonged use of systemic corticosteroids. These patients may benefit from treatment with anti-IgE, after a meticulous reevaluation of possible reasons for the failure to control asthma. PMID- 17136289 TI - Treatment of gastroesophageal reflux disease. AB - OBJECTIVES: To review the literature on the treatment of gastroesophageal reflux disease (GERD) with emphasis on pharmacological aspects. To identify particularities of pharmacological treatment of esophageal and extraesophageal manifestations of the disease. SOURCES: Electronic search of the PubMed/MEDLINE and Cochrane Collaboration databases. Controlled and randomized studies published since 2000 and reviews representing consensus positions and directives published within the last 10 years were identified. SUMMARY OF THE FINDINGS: The drugs currently available for the treatment of GERD do not act in the primary mechanism of the disease, i.e. transitory relaxation of the lower esophageal sphincter. Pharmacological treatment of GERD with symptoms or with esophageal injury is based on the suppression of acid secretion, particularly with proton pump inhibitors. When the hyperreactivity of the lower airways coexists with esophageal GERD symptoms, suppression of acid secretions should be of benefit in managing the respiratory disease in the presence of a causal relationship; however, this is not usual. When esophageal symptoms are not present, esophageal 24-hour pH study should be carried out prior to starting pharmacological treatment for GERD. Improvement of respiratory symptoms may be delayed with relation to esophageal symptoms. It is common for GERD to recur and pharmacological treatment should be repeated or continued indefinitely, depending on clinical presentation of the disease. CONCLUSIONS: The strategies that have been proposed for the pharmacological treatment of GERD in children are primarily based on studies of case series or on studies with adults. There have been very few controlled and randomized studies in children. Undertaking a greater number of these studies might reinforce existing aspects or establish new aspects of management. PMID- 17136290 TI - Outpatient antibiotic therapy as a predisposing factor for bacterial resistance: a rational approach to airway infections. AB - OBJECTIVES: To present evidence-based recommendations for the use of antibiotics for the treatment of the most common acute respiratory infections (ARI) and the available information on the importance of this type of management. SOURCES: MEDLINE and LILACS databases, technical publications by international organizations, national and international directives. The search terms acute respiratory infection, otitis, sinusitis, tonsillitis, pneumonia, antibiotic, guidelines and bacterial resistance were used. Articles cited by the articles selected were analyzed for information of interest. SUMMARY OF THE FINDINGS: Bacterial resistance has grown, to the extent that today it is recognized as a global public health problem. ARI are the most common cause of antibiotic usage within the community; yet a large proportion of these cases, compromising the upper (otitis, sinusitis, tonsillitis) or the lower airways (pneumonia), are the result of viral infections. Recommendations to rationalize the use of antibiotics in patients with ARI have the common objective of minimizing unnecessary antibiotic use, since "antibiotic pressure" is one of the factors triggering bacterial resistance. CONCLUSIONS: It is of great importance to differentiate among ARI patients those who will benefit from the use of antibiotics. The establishment of recommendations for the prescription of antibiotics is one strategy for minimizing the frequency of bacterial resistance. PMID- 17136291 TI - Treatment of pulmonary arterial hypertension. AB - OBJECTIVE: To perform a review of the diagnostic and therapeutic management of pulmonary hypertension in the pediatric population, with emphasis on pharmacological factors. SOURCES: Electronic search of publications on the MEDLINE/PubMed, LILACS and Cochrane Collaboration databases. The search strategy adopted gave priority to the identification of clinical trials (controlled or uncontrolled), systematic reviews and directives published during the last 10 years. SUMMARY OF THE FINDINGS: Many advances have been incorporated into our understanding of pulmonary hypertension during recent years. Issues related to differences in the pathophysiological mechanism of the disease between different age groups have altered both the treatment and prognosis of patients. The combined effect of more selective vasodilatory properties and antiproliferative action and the employment of new drugs are the basic principles of new treatment proposals. In order to be able to gauge the benefits associated with the use of these new therapies, it is of fundamental importance that all patients have their disease correctly diagnosed, the degree of functional compromise classified and their vascular reactivity capacity established, which is more difficult with pediatric patients. CONCLUSIONS: To date there is no treatment that can be considered ideal for the management of pulmonary hypertension. With reference to the possibility of employing new drugs, the majority of studies that have been published were undertaken with adult populations. Few data are available on children, and the majority of studies are uncontrolled trials or case series. Taking into account differences that have already been established between different age groups in terms of disease mechanisms and prognostic aspects, it is difficult to claim that these drugs can be incorporated into the treatment of childhood pulmonary hypertension with the same indications and results. PMID- 17136292 TI - Calcineurin inhibitors in the treatment of allergic dermatitis. AB - OBJECTIVE: To review the role of calcineurin inhibitors in the treatment of allergic dermatitis, focusing on mechanisms of action, efficacy and topical and systemic adverse effects. SOURCES: Articles written in English and published in MEDLINE using the following keywords: pimecrolimus, tacrolimus, calcineurin inhibitors. Original articles that presented controlled and open studies for assessing efficacy, tolerability and adverse effects were selected. Review articles and case series were also evaluated; the latter was only considered for assessing adverse effects. The official websites of the Food and Drug Administration and of manufacturers of calcineurin inhibitors were also used. SUMMARY OF THE FINDINGS: The data showed that calcineurin inhibitors are efficient in the treatment of mild to severe atopic dermatitis, leading to improvement in symptoms, reduction in number of attacks and need of topical corticotherapy. Calcineurin inhibitors have good tolerability and few topical adverse effects. To date, there has been no evidence to support higher prevalence of neoplasia in patients using these drugs; however, an adequate pharmacovigilance system has been set up to assess this aspect. CONCLUSIONS: Calcineurin inhibitors, which are a new drug class in the treatment of allergic dermatitis, are efficient, well tolerated and have few adverse effects. Calcineurin inhibitors should always be used according to recommended guidelines, and patients should always be followed by the physician during and after their administration. PMID- 17136293 TI - New antihistamines: a critical view. AB - OBJECTIVES: To perform a critical evaluation of the more recent H1 antihistamines and the various terms used to describe them, based on a review of evidence on their role in the treatment of allergic disorders. SOURCES OF DATA: Original articles, reviews and consensus documents published from 1998 to 2006 and indexed in the MEDLINE and PubMed databases. Keyword: antihistamines. SUMMARY OF THE FINDINGS: Second-generation antihistamines differ from first-generation ones because of their elevated specificity and affinity for peripheral H1 receptors and because of their lower penetration of the central nervous system (CNS), having fewer sedative effects as a result. Whilst second-generation antihistamines are in general better tolerated than their predecessors, some adverse effects, principally cardiotoxicity, have been observed with some of them. Over the last 20 years, new compounds with different pharmacokinetic properties have been synthesized. The majority of these exhibit anti-inflammatory properties that are independent of their action on the H1 receptor. More recent improvements, generally in the form of active metabolites, led to the use of the term third-generation antihistamines. This term emerged spontaneously, with no clear definition of its meaning or clinical implications, creating great confusion among healthcare professionals. CONCLUSION: On the basis of the evidence on H1 antihistamines, none of them deserve the title "third-generation antihistamine." As the Consensus Group on New Generation Antihistamines concluded, to merit this definition, a new class of antihistamines would have to demonstrate distinct clinical advantages over existing compounds and fulfill at least three prerequisites: they should be free from cardiotoxicity, drug interactions and effects on the CNS. PMID- 17136294 TI - Allergy to beta-lactams in pediatrics: a practical approach. AB - OBJECTIVE: To present a practical approach to the diagnosis and management of allergy to beta-lactam antibiotics. SOURCES: Allergy journals indexed in MEDLINE and LILACS, as well as seminal studies and texts. SUMMARY OF THE FINDINGS: Allergy to penicillin is commonly reported. In many cases, this results in the decision not to use this drug. About 10% of drug allergy reports are confirmed. The clinical manifestations due to allergic reaction to penicillin vary widely, with emphasis on skin disorders. Gell & Coombs' four hypersensitivity mechanisms are involved in allergic reactions. Penicillin is degraded to a major (95%) and minor determinants (5%). Immediate IgE-mediated reactions causing anaphylaxis are associated with minor determinants in 95% of the cases. Hypersensitivity to these products can be assessed using cutaneous tests performed with major and minor determinants, thus avoiding anaphylactic shock in allergic individuals. The present article underscores the basic body of knowledge on allergy to penicillin, providing support for a more accurate diagnosis of this event and for the choice of management in cases of suspected beta-lactam allergy. CONCLUSIONS: The incorrect diagnosis of penicillin allergy frequently leads to the exclusion of this drug as a therapeutic option. A better recognition of these situations will enable the use of penicillin and reduce the risks associated with hypersensitivity. PMID- 17136295 TI - The role of probiotics and prebiotics in pediatric practice. AB - OBJECTIVE: To review the effects of probiotics and prebiotics in clinical pediatric practice. SOURCES: MEDLINE was searched, especially for articles that addressed their practical application, in the form of reviews, clinical trials and meta-analyses. Articles that had already been analyzed by the authors were also included. SUMMARY OF THE FINDINGS: Scientific literature on probiotics and prebiotics has remarkably increased in the last 10 years. Their mechanisms of action have been experimentally investigated. Studies indicate that probiotics can act by competing with pathogens, modifying the intestinal environment by reduction in pH, as a result of fermentation products, interacting and modulating local and systemic inflammatory and immune response, among others. Clinical trials and meta-analyses show that probiotics seem to contribute towards the prevention of acute diarrhea and of antibiotic-associated diarrhea, in addition to shortening the duration of acute diarrhea. However, the data are inconsistent and there are no studies confirming their efficacy in terms of cost-benefit ratio. Preliminary studies show that probiotics in early life can reduce the occurrence of atopic dermatitis. The addition of prebiotics to infant formulas is associated with the change in the profile of the intestinal microbiota compared to infants fed milk formulas without prebiotics. CONCLUSIONS: Evidence indicates that new studies should be carried out about probiotics, prebiotics and symbiotics. The specific clinical effects that each probiotic or prebiotic may cause must be considered. PMID- 17136296 TI - Inhaled corticosteroids in the treatment of respiratory allergy: safety vs. efficacy. AB - OBJECTIVE: Review the molecular mechanisms of action, efficacy, and potential side effects associated with inhaled corticosteroids (ICS) in children with persistent asthma. SOURCES: Articles in English from MEDLINE. The following terms were used: corticosteroids, inhaled corticosteroids, asthma, children, beclomethasone, fluticasone, budesonide, ciclesonide, growth, adrenal insufficiency, bone mineral density, and oral candidiasis. Treatment guidelines, review articles, controlled trials, meta-analyses, and systematic reviews evaluating the efficacy and the adverse events of treatment with ICS were selected. SUMMARY OF THE FINDINGS: In vivo and in vitro studies show that the available ICS have different pharmacokinetic and pharmacodynamic properties that result in different action potentials. ICS also differ as to the systemic and local side effects. The bioavailability of these products is essential in order to determine the incidence of side effects. In general, ICS are capable of controlling asthma, reducing the number of exacerbations, medical consultations, hospitalizations, and the need of oral corticosteroid (applications) bursts. Improvement can also be seen in pulmonary function, especially in patients with recent onset asthma. The most documented adverse effect is transitory decrease of growth rate. CONCLUSIONS: ICS are the main anti-inflammatory agent used to treat persistent asthma. When administered in low doses, they seem to be safe and effective. Patient monitoring allows for early detection of possible side effects associated with ICS. PMID- 17136297 TI - Nonsteroidal anti-inflammatory drugs: cyclooxygenase 2 inhibitors. AB - OBJECTIVES: To analyze selective COX 2 inhibitor nonsteroidal anti-inflammatory drugs (NSAID) in terms of their mechanism of action, principal indications, posology and most common adverse effects. SOURCES: MEDLINE and LILACS databases and Food and Drug Administration (FDA) and National Agency for Sanitary Vigilance (ANVISA - Agencia Nacional de Vigilancia Sanitaria) websites. The most important articles were selected and preference was given to articles published within the last 5 years. SUMMARY OF THE FINDINGS: The principal indications for NSAID are for control of pain and acute and chronic inflammation. There is no overwhelming evidence that demonstrates the superiority of one NSAID over another in terms of effectiveness. To date none of the COX 2 inhibitors has been liberated for use in the pediatric age group. Only meloxicam and etoricoxib can be prescribed for adolescents (13 and 16 years, respectively). Selective COX 2 inhibitors are indicated for patients with adverse effects that have proven to be associated with nonselective NSAID use. Selective COX 2 inhibitors can be prescribed in some cases of allergy to aspirin, but they must be used with care. Principal adverse effects include cardiovascular events and thrombotic phenomena. CONCLUSIONS: Selective COX 2 inhibitors are medicines that have been used in certain well defined clinical situations and which may offer certain advantages over nonselective NSAID. Nevertheless, taking into consideration the higher cost involved and the potential for adverse cardiovascular effects, they should be employed only in accordance with strict criteria. PMID- 17136298 TI - Antileukotrienes in the treatment of asthma and allergic rhinitis. AB - OBJECTIVES: To compare leukotriene antagonists (LTA) to other groups of drugs used in asthma and allergic rhinitis treatment. SOURCES: MEDLINE, LILACS and Cochrane Library. KEYWORDS: leukotrienes, antileukotrienes, asthma treatment, allergic rhinitis treatment, asthma and allergic rhinitis. An attempt was made to group the main studies and reviews about this topic. SUMMARY OF THE FINDINGS: LTA are more efficient than placebo and enhance the effects of inhaled corticosteroids. The association of inhaled corticosteroids with long-acting Beta2 agonists is more efficient than the association of inhaled corticosteroids + LTA. Although use of LTA in acute asthma attacks and allergic rhinitis seems reasonable, more studies are needed to confirm this benefit. LTA reduce hospitalization time and the number of wheezing attacks in infants with acute viral bronchiolitis caused by respiratory syncytial virus, as well as recurrent wheezing after acute viral bronchiolitis. LTA are less efficient than intranasal corticosteroids for allergic rhinitis management. LTA are efficient in exercise induced asthma, although they are not the first-line treatment. CONCLUSION: Controlled and randomized studies show that inhaled corticosteroids are the drugs of choice to treat persistent asthma and allergic rhinitis. There is not enough evidence to recommend the use of LTA as first-line drug (monotherapy) in children with asthma (level I). For children who cannot use inhaled corticosteroids, LTA may be a good alternative (level II). PMID- 17136299 TI - Temperament, life events, and psychopathology among the offspring of bipolar parents. AB - OBJECTIVES: The present study examines the relationship between temperament, recent and remote life events, and psychopathology among the offspring of parents with bipolar disorder and well comparisons. METHODS: Offspring of bipolar and well parents were clinically assessed using KSADS-PL format interviews. Lifetime psychiatric diagnoses were made on a blind consensus basis in accordance with DSM IV criteria. Depending on offspring age, either the child or their parent on their behalf, completed a semi-structured interview quantifying the number and impact of recent life events and remote permanent losses, as well as a measure of temperament. RESULTS: In this study, there was an association between psychopathology and the number of recent negative life events, but no association between psychopathology and the number of early losses. Emotionality was positively correlated with recent life events. However, in stepwise regression analyses, only emotionality significantly contributed to lifetime psychopathology in general and emotionality and age contributed to the risk of mood disorder in particular. CONCLUSIONS: These findings, replicate in a sample of offspring at high risk for bipolar disorder, previously reported associations between high emotionality and unipolar depression. In this population, any effect of undesirable life events would appear to be mediated through the association with emotionality. PMID- 17136300 TI - Adaptation and adjustment in children of transsexual parents. AB - It is a relatively uncommon event that children find themselves in a situation with a parent who undergoes a transition from one sex to another. Unlike situations of divorce or a parent with a medical or major psychiatric disorder, it is unlikely that the children will know of other children who encounter similar situations. There is very little literature that describes the adjustment of these children and the nature of their relationships with their parents and peers. Such information would be beneficial for clinicians assisting children and families in this situation. To better delineate the adjustment of these children, we interviewed 27 parents of 55 children. The interview took place on average 6 years after the gender transition. The measures obtained included the parent and child relationships at present and also at the time of the transition. We also inquired about academic function, peer relationships and social stigma. The results found that children who were younger at the time of the parent's transition tended to have better relationships and less adjustment difficulties. In addition, parental conflict that continues after the transition period tends to reflect greater family conflict between the transitioned parent and their child. PMID- 17136301 TI - Can autism, language and coordination disorders be differentiated based on ability profiles? AB - Children with autistic disorder (AD), mixed receptive-expressive language disorder (RELD), or developmental coordination disorder (DCD) have impairments in common. We assess which abilities differentiate the disorders. Children aged 3-13 years diagnosed with AD (n = 30), RELD (n = 30), or DCD (n = 22) were tested on measures of language, intelligence, social cognition, motor coordination, and executive functioning. Results indicate that the AD and DCD groups have poorer fine and gross motor coordination and better response inhibition than the RELD group. The AD and DCD groups differ in fine and gross motor coordination, emotion understanding, and theory of mind scores (AD always lower), but discriminant function analysis yielded a non-significant function and more classification errors for these groups. In terms of ability scores, the AD and DCD groups appear to differ more in severity than in kind. PMID- 17136302 TI - Visual completion and complexity of visual shape in children with pervasive developmental disorder. AB - Much evidence has been gathered for differences in visual perceptual processing in individuals with Autistic Spectrum Disorder. The presence of the fundamental process of visual completion was tested in a group of children with Pervasive Developmental Disorder (PDD), as this requires perceptually integrating visual structure into wholes. In Experiment 1, it was investigated whether visual completion is present for simple partly occluded shapes in a group of children with PDD and a typically developing group. In Experiment 2, the presence of contextual influences in visual completion was investigated for the two groups. A total of 19 children with PDD and 28 controls who were matched for chronological age and IQ took part in two primed-matching tasks. For both groups, visual completion was present and for both groups, contextual influences were found to be dominant in this process. However, only for the group with PDD no priming effects (PEs) were found from less regular primes on congruent test pairs. The group with PDD did integrate visual information into wholes and did this in a contextually dependent way. However, for more complex shapes, visual completion is weaker for this group. PMID- 17136303 TI - Attention deficit hyperactivity disorder among Nigerian primary school children: prevalence and co-morbid conditions. AB - OBJECTIVE: This study aimed to determine the prevalence of ADHD and co-morbid conditions in a sample of primary school children aged 7-12 years in Nigeria. METHOD: A two-staged procedure in which primary school pupils aged 6-12 years (n = 1112) were assessed for DSM-IV criteria of attention deficit hyperactivity disorder (ADHD) by their teachers in the first stage and their parents in the second stage. A flexible criterion was used for estimating the prevalence. RESULTS: The prevalence of ADHD was 8.7%. The prevalence of the subtypes were: predominantly Inattentive 4.9%, predominantly hyperactive/impulsive 1.2% and combined 2.6%. The male to female ratio was 2:1 for all the subtypes of ADHD except hyperactive/impulsive which was 3.2:1. The co-morbid conditions include oppositional defiant disorder (ODD - 25.8%), conduct disorder (CD - 9.3%) and anxiety/depression (20.6%). While ODD and CD were associated with the hyperactive/impulsive subtype, anxiety/depression was associated with inattentive subtype. CONCLUSION: Our findings support the notion that ADHD occurs across cultures. Given the prevalent rate, efforts should be made to map out strategies for early identification and referral of these children for proper evaluation and treatment. This study can serve as a platform for future analytical studies about this challenging research issue in sub-Saharan Africa. PMID- 17136304 TI - Expression of hepatitis B surface antigen in Saccharomyces cerevisiae utilizing glyceraldeyhyde-3-phosphate dehydrogenase promoter of Pichia pastoris. AB - An expression vector constructed from genes of Pichia pastoris was applied for heterologous gene expression in Saccharomyces cerevisiae. Recombinant hepatitis B surface antigen was synthesized by cloning hepatitis B virus 'S' gene under the control of glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter of Pichia pastoris in Saccharomyces cerevisiae. Hepatitis B surface antigen was constitutively expressed, was stable and exhibited approximately 20-22 nm particle formation. Stability and absence of toxicity to the host with the expression vector indicates the expression system can be applied for large-scale production. PMID- 17136305 TI - Isolated and characterization of a cDNA encoding ethylene-responsive element binding protein (EREBP)/AP2-type protein, RCBF2, in Oryza sativa L. AB - A transcription factor RCBF2 which interacts with C-repeat/DRE was isolated from Oryza sativa L. by a yeast one-hybrid method. Analysis of the deduced RCBF2 amino acid sequence revealed that RCBF2 contained a conserved ethylene-responsive element binding protein (EREBP)/AP2 domain of 59 amino acids and a potential nuclear localization sequence. RCBF2 showed a high level of homology with other CBF family members only in AP2 domain. Phylogenetic analysis showed that RCBF2 might be different from other eight DRE-binding proteins on evolutionary relationship. The semi-quantitative RT-PCR (s-Q RT-PCR) analysis indicated the expression of RCBF2 gene was induced by cold, dehydration and high-salinity, but not by abscisic acid, and the transcription of RCBF2 gene accumulated primarily in rice immature seeds, growing point and shoots. PMID- 17136306 TI - Fatty acids reduce the tensile strength of fungal hyphae during cephalosporin C production in Acremonium chrysogenum. AB - Fragmentation rate constants, which can be used to estimate the tensile strength of fungal hyphae, were used to elucidate relationships between morphological changes and addition of fatty acids during cephalosporin C production in Acremonium chrysogenum M35. The number of arthrospores increased gradually during fermentation, and, in particular, was higher in the presence of rice oil, oleic acid or linoleic acid than in their absence. Because supplementation of rice oil or fatty acids increased cephalosporin C, we concluded that differentiation to arthrospores is related to cephalosporin C production. To estimate the relative tensile strengths of fungal hyphae, fragmentation rate constants (k (frag)) were measured. When rice oil, oleic acid, or linoleic acid were added into medium, fragmentation rate constants were higher than for the control, and hyphal tensile strengths reduced. The relative tensile strength of fungal hyphae, however was not constant presumably due to differences in physiological state. PMID- 17136307 TI - Subcutaneous emphysema: a rare manifestation of a perforated diverticulitis in a patent inguinal canal. AB - Patients with complicated diverticulitis rarely present with extraperitoneal manifestations but the manifestation of subcutaneous emphysema appears even more seldom. We present the case of a patient with a history of diabetes and immunosuppression, who was admitted with sepsis in association with cellulitis and subcutaneous emphysema of the left groin. The absence of peritonism due to corticosteroid treatment, a history of a recent fall with an ilio- and ischio pubic fracture and subcutaneous emphysema led to a delay in the diagnosis. The final diagnosis was a perforated diverticulitis in a patent inguinal canal, which was only revealed after surgery. The various complications of diverticulitis, including extraperitoneal manifestations, and associated microorganisms implicated in cellulitis and subcutaneous emphysema are briefly reviewed. PMID- 17136308 TI - Giant spigelian hernia due to abdominal wall injury: a case report. AB - Spigelian hernia is a rare clinical entity. It is difficult to diagnose due to its location. In this article we report the case of a giant spigelian hernia consequent to abdominal wall injury. The neck of the hernia was 10 cm in diameter. We repaired this hernia with a polypropylene mesh. PMID- 17136309 TI - A prospective comparison of local and spinal anesthesia for inguinal hernia repair. AB - AIM: Today, in inguinal hernia repair, postoperative pain and costs are regarded as equally important issues as technique and recurrence rates. Postoperative pain is thought to vary according to the applied anesthesia method. As local anesthesia is reported to inflict less pain, its effects on early period post operative complications should also be evaluated. METHODS: Two hundred patients, on whom Lichtenstein tension free hernia repair had been performed due to unilateral inguinal hernia between March 2004 and July 2005, were prospectively examined. The patients were randomized according to the anesthesia applied. They were divided into two groups: local anesthesia (LA) and spinal anesthesia (SA). The early post-operative complications, post-operative pain scores, and operation durations of the patients, were evaluated. RESULTS: Local anesthesia was found not to increase the post-operative complications; on the contrary, it was shown to prevent the complications of spinal anesthesia. Although visual analogue pain score (VAS) values at 4, 8, 12, and 24 h post-operation were found to be lower than the SA group, the difference between was not significant. Also, it was discovered that LA did not retard the operation duration. CONCLUSION: Local anesthesia reduces post-operative pain and facilitates patients' mobilization and discharge along with decreasing the early post-operative complications. Thus, LA is a safe and advantageous method to be applied in inguinal hernia repair. PMID- 17136310 TI - Cadmium induces nuclear translocation of beta-catenin and increases expression of c-myc and Abcb1a in kidney proximal tubule cells. AB - Cadmium (Cd2+) induces renal proximal tubular (PT) damage, including disruption of the E-cadherin/beta-catenin complex of adherens junctions (AJs) and apoptosis. Yet, chronic Cd2+ exposure causes malignant transformation of renal cells. Previously, we have demonstrated that Cd(2+)-mediated up-regulation of the multidrug transporter Abcb1 causes apoptosis resistance in PT cells. We hypothesized that Cd2+ activates adaptive signaling mechanisms mediated by beta catenin to evade apoptosis and increase proliferation. Here we show that 50 microM Cd2+, which induces cell death via apoptosis and necrosis, also causes a decrease of the trans-epithelial resistance of confluent WKPT-0293 Cl.2 cells, a rat renal PT cell model, within 45 min of Cd2+ exposure, as measured by electric cell-substrate impedance sensing. Immunofluorescence microscopy demonstrates Cd(2+)-induced decrease of E-cadherin at AJs and redistribution of beta-catenin from the E-cadherin/beta-catenin complex of AJs to cytosol and nuclei after 3 h. Immunoblotting confirms Cd(2+)-induced decrease of E-cadherin expression and translocation of beta-catenin to cytosol and nuclei of PT cells. RT-PCR shows Cd(2+)-induced increase of expression of c-myc and of the isoform Abcb1a at 3 h. The data prove for the first time that Cd2+ induces nuclear translocation of beta catenin in PT cells. We speculate that Cd2+ activates beta-catenin/T-cell factor signaling to trans-activate proliferation and apoptosis resistance genes and promote carcinogenesis of PT cells. PMID- 17136311 TI - Heterozygous tx mice have an increased sensitivity to copper loading: implications for Wilson's disease carriers. AB - Wilson's disease carriers constitute 1% of the human population. It is unknown whether Wilson's disease carriers are at increased susceptibility to copper overload when exposed to chronically high levels of ingested copper. This study investigated the effect of chronic excess copper in drinking water on the heterozygous form of the Wilson's disease mouse model--the toxic milk (tx) mouse. Mice were provided with drinking water containing 300 mg/l copper for 4-7, 8-11, 12-15 or 16-20 months. At the completion of the study liver, spleen, kidney and brain tissue were analyzed by atomic absorption spectroscopy to determine copper concentration. Plasma ceruloplasmin oxidase activity and liver histology were also assessed. Chronic copper loading resulted in significantly increased liver copper in both tx heterozygous and tx homozygous mice, while wild type mice were resistant to the effects of copper loading. Copper loading effects were greatest in tx homozygous mice, with increased extrahepatic copper deposition in spleen and kidney - an effect absent in heterozygote and wild type mice. Although liver histology in homozygous mice was markedly abnormal, no histological differences were noted between heterozygous and wild type mice with copper loading. Tx heterozygous mice have a reduced ability to excrete excess copper, indicating that half of the normal liver Atp7b copper transporter activity is insufficient to deal with large copper intakes. Our results suggest that Wilson's disease carriers in the human population may be at increased risk of copper loading if chronically exposed to elevated copper in food or drinking water. PMID- 17136312 TI - Hypernatremic myopathy caused by a hypothalamic mixed germ cell tumor mimicking polymyositis. AB - Hypernatremic myopathy was rarely reported in the literature and its clinical features have never been well-described. We present a 22-year-old man who had adipsic hypernatremia manifested with progressive proximal muscle weakness and remarkably high creatine kinase level that has never been reported in the cases of hypernatremic myopathy. His initial presentations were similar to that of polymyositis without the evidence of central nervous system dysfunction and hypopituitarism. The serum level of sodium at the beginning of myopathy is the lowest known in the literature. All the clinical presentations in this patient resulted from a hypothalamic mixed germ cell tumor with sub-acute intra-tumoral hemorrhage. PMID- 17136313 TI - Absence of antibody to cyclic citrullinated peptide in sera of non-arthritic patients with chronic hepatitis B virus infection. AB - The objective of this study was to investigate if antibody to cyclic citrullinated peptide (anti-CCP) is detected in sera of patients with chronic hepatitis B virus (HBV) infection. Serum anti-CCP and IgA, IgG, and IgM rheumatoid factor (RF) isotypes were measured by enzyme-linked immunosorbent assay on 176 non-arthritic patients with HBV infection. IgA RF, IgG RF, and IgM RF were detectable in 29.5, 21, and 18.8% of the tested sera, respectively, with a total seropositivity rate of 42.7%. Marginally elevated anti-CCP was detected in one patient (0.6%). By regression analysis, there was no statistically significant association between the serum levels of anti-CCP and serum IgA, IgG, or IgM RF (R (2) = 0.033, with respective p values of 0.224, 0.297, and 0.334). In conclusion, anti-CCP was rarely detected in non-arthritic patients with HBV infection in contrast to RF. Thus, testing for anti-CCP may be a useful tool for the diagnosis of rheumatoid arthritis in this population. PMID- 17136315 TI - Numerical model study of flow dynamics through an end-to-side anastomosis: choice of anastomosis angle and prosthesis diameter. AB - The purpose of this numerical model study was to determine the angle of anastomosis and prosthesis-to-artery diameter ratio that theoretically limits development of myointimal hyperplasia (MIH). Blood flow patterns were investigated in a model simulating a distal end-to-side anastomosis on a 2-mm diameter artery. Tests were carried out under steady and pulsatile flow conditions with and without taking into account the non-Newtonian behavior of blood and compliance. The wall shear stress gradient (WSSG), a potential factor for development of MIH, was analyzed as a function of the angle of anastomosis (18 degrees , 25 degrees , 35 degrees , and 45 degrees ) and prosthesis diameter (4, 5, and 6 mm). The angle of anastomosis that minimized WSSG was 18 degrees . Prosthesis diameter had no effect on WSSG, with similar results for all three diameters. These findings suggest that surgeons should choose as acute an angle of anastomosis as possible. Prosthesis diameter played no role in reducing WSSG values. PMID- 17136316 TI - Preliminary examination of short-term cellular toxicological responses of the coral Madracis mirabilis to acute Irgarol 1051 exposure. AB - Irgarol 1051 is an s-triazine herbicide formulated with Cu2O in antifouling paints. Recent studies have shown that Irgarol 1051 inhibits coral photosynthesis at environmentally relevant concentrations, consistent with its mode of action as a photosystem II inhibitor. Related toxicologic effects of this herbicide on coral cellular physiology have not yet been investigated. We used cellular diagnostics to measure changes in 18 toxicologic cellular parameters in endosymbiotic algal (dinoflagellate) and cnidarian (host) fractions of the common branching coral Madracis mirabilis associated with in vivo 8- and 24-hour exposures to a nominal initial Irgarol 1051 concentration of 10 microg L(-1). Responses measured were (1) xenobiotic response, which includes total and dinoflagellate multixenobiotic resistance (MXR), cnidarian cytochrome (CYP) P450 3 and P450-6 classes, cnidarian, and dinoflagellate glutathione-s-transferase (GST); (2) oxidative damage and response, which includes cnidarian and dinoflagellate Cu/Zn and Mn superoxide dismutase (SOD), cnidarian and dinoflagellate glutathione peroxidase (GPx), cnidarian catalase, and total protein carbonyl); (3) metabolic homeostasis, which includes chloroplast and invertebrate small heat-shock proteins (sHsp), cnidarian protoporphyrinogen oxidase IX (PPO), cnidarian ferrochelatase, and cnidarian heme oxygenase; and (4) protein metabolic condition, which includes cnidarian and dinoflagellate heat shock proteins (hsp70 and hsp60), total ubiquitin, and cnidarian ubiquitin ligase. Acute responses to Irgarol 1051 exposure included significant increases in total and dinoflagellate MXR, dinoflagellate Cu/Zn SOD, dinoflagellate chloroplast sHsp, and cnidarian PPO. Irgarol 1051 exposure resulted in decreases in cnidarian GPx, cnidarian ferrochelatase, cnidarian catalase, and cnidarian CYP 450-3 and -6 classes. Related implications of Irgarol 1051 exposure to coral cellular condition are discussed. PMID- 17136317 TI - Desulfomicrobium thermophilum sp. nov., a novel thermophilic sulphate-reducing bacterium isolated from a terrestrial hot spring in Colombia. AB - A moderately thermophilic, sulphate-reducing bacterium, designated strain P6 2(T), was isolated from a terrestrial hot spring located at a height of 2,500 m in the Andean region, Colombia (5 degrees 43'69''N, 73 degrees 6'10''W). Cells of strain P6-2(T) were rod-shaped, stained Gram-negative and were motile by means of a single polar flagellum. The strain grew lithotrophically with H(2) as the electron donor and organotrophically on lactate, pyruvate, ethanol, malate, fumarate, n-propanol and succinate in the presence of sulphate as the terminal electron acceptor. Fumarate and pyruvate was fermented. Strain P6-2(T) grew optimally at 55 degrees C (range 37-60 degrees C), pH 6.6 (range 5.8-8.8) in the presence of 0.5% NaCl (range 0-4.5%) with lactate and sulphate and produced acetate, CO(2) and H(2)S as the major end-products. Sulphate, sulphite and thiosulphate could be used as electron acceptors but not elemental sulphur or nitrate. The G + C content of the genomic DNA was 58.7 mol%. The 16S rRNA sequence analysis indicated that strain P6-2(T) was a member of the class Deltaproteobacteria, domain Bacteria with Desulfomicrobium baculatum being the closest relative (similarity value of 94%). Phylogeny of genes encoding alpha- and beta-subunits of the dissimilatory sulphite reductase (dsrAB genes) supported its affiliation to members of the genus Desulfomicrobium. On the basis of this evidence, we propose to assign strain P6-2(T) as new species of the genus Desulfomicrobium, D. thermophilum sp. nov., with strain P6-2(T) as the type strain (= DSM 16697(T) = CCUG 49732(T)). PMID- 17136318 TI - Comparison of the rapid pro-apoptotic effect of trans-beta-nitrostyrenes with delayed apoptosis induced by the standard agent 5-fluorouracil in colon cancer cells. AB - Trans-beta-nitrostyrene (TBNS) has been reported to be a potent inhibitor of protein phosphatases PTB1 and PP2A and to display a pro-apoptotic effect even in multidrug resistant tumour cells. Here we compared the anti-tumour potential of TBNS with 5-fluorouracil (5-FU) as the standard chemotherapeutic agent for colorectal cancer in LoVo cells. Resistance to 5-FU based therapy might be a consequence of 5-FU's delayed effect requiring long-term effective concentrations in the tumour tissue. Thus, alternatives like platin containing drugs with a more rapid effect have been introduced recently. Compared to 5-FU TBNS displayed a faster cytotoxic and pro-apoptotic effect. A 50% decrease in viability was observed already after 8 h with TBNS while 5-FU displayed no significant effect before 48 h. DNA fragmentation and caspase-3 assays confirmed the more rapid apoptotic effect of TBNS. Since apoptosis affects individual cells these results about a rapidly induced apoptosis were further studied on a single cell level in microscopic assays of caspase-3 and caspase-8 activation. Adducts of trans-beta nitrostyrene displayed an anti-tumour effect comparable to TBNS which suggests the possibility of creating adducts with optimised tissue targeting. Finally, the calculation of a drug combination index displayed a synergistic effect for the combination of TBNS and 5-FU in Lovo as well as in HT-29 and HCT116 colon cancer cells. PMID- 17136319 TI - Down-regulation of cFLIP following reovirus infection sensitizes human ovarian cancer cells to TRAIL-induced apoptosis. AB - Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) shows promise as a chemotherapeutic agent. However, many human cancer cells are resistant to killing by TRAIL. We have previously demonstrated that reovirus infection increases the susceptibility of human lung (H157) and breast (ZR75-1) cancer cell lines to TRAIL-induced apoptosis. We now show that reovirus also increases the susceptibility of human ovarian cancer cell lines (OVCAR3, PA-1 and SKOV-3) to TRAIL-induced apoptosis. Reovirus-induced increases in susceptibility of OVCAR3 cells to TRAIL require virus uncoating and involve increased activation of caspases 3 and 8. Reovirus infection results in the down-regulation of cFLIP (cellular FLICE inhibitory protein) in OVCAR3 cells. Down-regulation of cFLIP following treatment of OVCAR3 cells with antisense cFLIP oligonucleotides or PI3 kinase inhibition also increases the susceptibility of OVCAR3 cells to TRAIL induced apoptosis. Finally, over-expression of cFLIP blocks reovirus-induced sensitization of OVCAR3 cells to TRAIL-induced apoptosis. The combination of reovirus and TRAIL thus represents a promising new therapeutic approach for the treatment of ovarian cancer. PMID- 17136320 TI - Sulindac-derived reactive oxygen species induce apoptosis of human multiple myeloma cells via p38 mitogen activated protein kinase-induced mitochondrial dysfunction. AB - Non-steroidal anti-inflammatory drugs are well known to induce apoptosis of cancer cells independent of their ability to inhibit cyclooxygenase-2, but the molecular mechanism for this effect has not yet been fully elucidated. The purpose of this study was to elucidate the potential signaling components underlying sulindac-induced apoptosis in human multiple myeloma (MM) cells. We found that sulindac induces apoptosis by promoting ROS generation, accompanied by opening of mitochondrial permeability transition pores, release of cytochrome c and apoptosis inducing factor from mitochondria, followed by caspase activation. Bcl-2 cleavage and down-regulation of the inhibitor of apoptosis proteins (IAPs) family including cIAP-1/2, XIAP, and survivin, occurred downstream of ROS production during sulindac-induced apoptosis. Forced expression of survivin and Bcl-2 blocked sulindac-induced apoptosis. Most importantly, sulindac-derived ROS activated p38 mitogen-activated protein kinase and p53. SB203580, a p38 mitogen activated protein kinase inhibitor, and RNA inhibition of p53 inhibited the sulindac-induced apoptosis. Furthermore, p53, Bax, and Bak accumulated in mitochondria during sulindac-induced apoptosis. All of these events were significantly suppressed by SB203580. Our results demonstrate a novel mechanism of sulindac-induced apoptosis in human MM cells, namely, accumulation of p53, Bax, and Bak in mitochondria mediated by p38 MAPK activation downstream of ROS production. PMID- 17136321 TI - Involvement of caspase-8 in chemotherapy-induced apoptosis of patient derived leukemia cell lines independent of the death receptor pathway and downstream from mitochondria. AB - Resistance of leukemic cells to chemotherapy frequently occurs in patients with acute leukemia, which may be caused by alterations in common apoptotic pathways. Controversy exists whether cytostatic agents induce the mitochondrial or death receptor pathway of apoptosis. In the mitochondrial pathway cytochrome C release and caspase-9 activation play a central role in the induction of apoptosis, while formation of a Death Inducing Signaling Complex (DISC) and caspase-8 activation have been reported to be essential in death receptor-induced apoptosis. Here, we show in human derived myeloid and lymphoblastic leukemia cell lines that caspase 8 plays a more important role than previously expected in apoptosis mediated via the mitochondrial pathway. We demonstrated in these malignant cells chemotherapy induced apoptosis independent of the death receptor pathway, since blocking this pathway using a retroviral construct encoding Flice inhibitory protein (FLIP) did not inhibit drug-induced apoptosis or caspase-8 activation, while overexpression of Bcl-2 completely inhibited both events. Furthermore, we showed that activation of caspase-8 by cytostatic agents occurred downstream from mitochondria. Since caspase-8 plays a central role in both death receptor- and chemotherapy-induced apoptosis of malignant cells from patients with acute leukemia, therapeutic strategies focusing at modulation and activation of caspase-8 may be successful in the treatment of drug-resistant malignancies. PMID- 17136322 TI - Role of the mitochondrial membrane permeability transition in cell death. AB - In recent years, the role of the mitochondria in both apoptotic and necrotic cell death has received considerable attention. An increase of mitochondrial membrane permeability is one of the key events in apoptotic or necrotic death, although the details of the mechanism involved remain to be elucidated. The mitochondrial membrane permeability transition (MPT) is a Ca(2+)-dependent increase of mitochondrial membrane permeability that leads to loss of Deltapsi, mitochondrial swelling, and rupture of the outer mitochondrial membrane. The MPT is thought to occur after the opening of a channel that is known as the permeability transition pore (PTP), which putatively consists of the voltage-dependent anion channel (VDAC), the adenine nucleotide translocator (ANT), cyclophilin D (Cyp D: a mitochondrial peptidyl prolyl-cis, trans-isomerase), and other molecule(s). Recently, significant progress has been made by studies performed with mice lacking Cyp D at several laboratories, which have convincingly demonstrated that Cyp D is essential for the MPT to occur and that the Cyp D-dependent MPT regulates some forms of necrotic, but not apoptotic, cell death. Cyp D-deficient mice have also been used to show that the Cyp D-dependent MPT plays a crucial role in ischemia/reperfusion injury. The anti-apoptotic proteins Bcl-2 and Bcl x(L) have the ability to block the MPT, and can therefore block MPT-dependent necrosis in addition to their well-established ability to inhibit apoptosis. PMID- 17136323 TI - Dopaminergic and adrenergic toxicities on SK-N-MC human neuroblastoma cells are mediated through G protein signaling and oxidative stress. AB - Dopamine and norepinephrine are neurotransmitters which participate in various regulatory functions of the human brain. These functions are lost in neurodegenerative diseases including Parkinson's disease and Alzheimer's disease. In this study, we used SK-N-MC neuroblastoma cells to investigate the cytotoxicities of high concentrations of dopamine and norepinephrine on neuronal cells. Dopamine, norepinephrine, as well as their corresponding synthetic agonists (SKF38393 and isoproterenol, respectively) triggered SK-N-MC cell death when applied at 50-100 muM persistently for 2 days. This catecholamine-induced cell death appears to be neuronal specific, as demonstrated by their inabilities of triggering apoptosis of A549 lung carcinoma cells and Cos-7 kidney fibroblasts. By pretreating SK-N-MC cells with target-specific inhibitors before administration of catecholamine, components of G protein signaling (i.e. G( s )/cAMP/PKA), monoamine oxidases, nitric oxide synthase, c-Jun N-terminal kinase and oxidative stress were found to be involved in this dopamine/norepinephrine induced cytotoxicity, which subsequently led to caspase-dependent and independent apoptotic responses as well as DNA degradation. In contrast, agonists of G( i )-coupled dopamine receptors and adrenergic receptors (quinpirole and UK14,304, respectively) were incapable of triggering apoptosis of SK-N-MC cells. Our results suggest that both G protein (G( s ))-mediated signaling cascade and oxidative stress participate in the dopamine/norepinephrine-induced neuronal apoptosis. PMID- 17136324 TI - Ursane triterpenoids inhibit atherosclerosis and xanthoma in LDL receptor knockout mice. AB - INTRODUCTION: In order to determine the mechanism of triterpenes, a class of secondary metabolites in plants, in modulating progression of vascular atherosclerotic lesions, we isolated three ursane triterpenoids (euscaphic acid, tormentic acid and 2alpha-hydroxyursolic acid) from aerial part of Salvia miltirrhiza Bge. and fed LDLr(-/-) mice the isolated compounds at a dose of 10 mg/kg p.o. for 24 weeks. MATERIALS AND METHODS: The treated mice were raised with a cholesterol-enriched (1.25%) diet. Implying serum and aorta MCP-1 analysis, we found that all mice treated with the compounds exhibited a significant reduction of whole body and vascular inflammation. RESULTS: The reduction of macrophage cells' number in aortic atherosclerotic lesions suggests that triterpenes treatment results in the development of a more stable plaque phenotype. Analysis of the structure-activity relationships demonstrates that compounds with a beta orientated hydrogen-bond forming group at C-3 exhibit more potent anti atherogenic effect than the alpha-counterpart on the development of atherosclerosis and xanthoma. However, the biological activities of the compounds are significantly reduced when they have C-19 hydrogen-bonds. CONCLUSION: These Results suggest that down-regulation of MCP-1 is the main mechanism for antiatherogenic activity of triterpenes and MCP-1 might play important roles in the development of atherosclerosis and xanthoma. PMID- 17136327 TI - Current topics in brain dopamine research: a tribute to Professor Oleh Hornykiewicz. PMID- 17136328 TI - Dopamine: from pharmacology to molecular biology and back. PMID- 17136329 TI - Viral vectors as a tool to model and treat Parkinson's disease. PMID- 17136330 TI - Imaging the role of dopamine in health and disease Parkinson's disease as a lesion model. PMID- 17136331 TI - [100 years of allergy: Clemens von Pirquet - his idea of allergy and its immanent concept of disease]. AB - Coming from his clinical research in the field of infectiology and immunology the Viennese pediatrician Clemens von Pirquet (1874-1929) introduced the term "allergy" in 1906. With it he wanted to describe in general a change in reactivity of the organism, namely in time, quality and quantity. In contrast to the widely accepted use of the word "allergy" today, where it is restricted to specific immunologic hypersensitivity reactions against harmless foreign antigens, allergy in Pirquets sense comprised as general term likewise increases and decreases of the reactivity and so both "hyper-" and "hyposensitivity reactions". In the context with the expansion of allergy to the human predisposition Pirquet emphasized, that the change of reactivity does not only depend on exogenous substances (so called allergens), but also on endogenous factors of the organism itself. - The orientation towards the organism and his reactivity is the central idea, which can be found in the complete works of Pirquet from 1903 to 1929 and which is presented in this publication for the first time. It is the true essence of his theory of allergy. PMID- 17136333 TI - Mandibular locking due to bilateral coronoid process hyperplasia. PMID- 17136332 TI - [Vegetarian nutrition: Preventive potential and possible risks. Part 1: Plant foods]. AB - Today vegetarian nutrition is more accepted and widespread in Europe than in former years. For a long time scientific research on vegetarian diets has focused mostly on malnutrition, whereas nowadays research centers increasingly on the preventive potential of plant-based diets. We followed a nutritive and a metabolic-epidemiological approach to obtain dietary recommendations. A MEDLINE research was performed for all plant food groups relevant for a vegetarian diet (key words: all relevant food groups, "vegetarian diet", "chronic disease", "cancer", "cardiovascular disease", "diabetes mellitus", "osteoporosis"). All relevant food groups were characterized regarding their nutrient content and rated with respect to the available metabolic-epidemiological evidence. Based on the evidence criteria of the WHO/FAO, cancer risk reduction by a high intake of vegetables and fruits is assessed as probable or possible, while a lowered risk of cardiovascular disease is convincing and a lowered risk of osteoporosis is probable. The evidence of a risk reducing effect of whole grain relating to colorectal cancer is assessed as possible, whereas it is probable relating to cardiovascular disease and diabetes mellitus type 2. There is an insufficient risk-reducing effect of legumes like soja relating to epithelial tumours and cardiovascular disease. The evidence of a risk-reducing effect of nuts to cardiovascular disease is assessed as probable, and in relation to cholelithiasis and diabetes mellitus type 2 as possible and insufficient, respectively. In conclusion, high consumption of fruits, vegetables, whole grains and nuts can lower the risk for several chronic diseases. PMID- 17136334 TI - Standard interferon-alpha in combination with ribavirin for hepatitis C patients with advanced liver disease and thrombocytopenia. AB - BACKGROUND AND AIM: Patients with advanced liver disease due to thrombocytopenia and chronic infection with hepatitis C virus (HCV) are difficult to treat in view of concerns about the efficacy and safety of interferon-based therapy. Nevertheless, antiviral therapy might have a substantial benefit in these patients as it potentially minimizes disease progression and prevents recurrence after liver transplantation. We evaluated the safety, efficacy and tolerability of standard interferon-alpha in an accelerating dose regimen in combination with ribavirin in patients with HCV-induced liver cirrhosis and thrombocytopenia. PATIENTS: Nine patients (M=8, age: 48.4 +/- 9.9, mean +/- SD) with HCV-related advanced liver disease and thrombocytopenia were prospectively investigated. The Child-Pugh stage was A in six patients and B in three, the MELD score was 11 [6 17] (median [range]). Four patients were interferon naive. HCV-genotype distribution was 1b (n=3), 3a (n=4) and 4 (n=2). The patients received 1-1.5 MU/d standard interferon-a2b with increasing dose regimen and weight-based ribavirin for 48 weeks (genotype 1), or 24 weeks (genotype 3), or until liver transplantation, respectively. RESULTS: The baseline platelet count was 64.3 +/- 8.7 (G/l, mean +/- SD) and remained remarkably stable during treatment (58.0 +/- 12.4 G/l at week 4, 51.7 +/- 20.5 G/l at week 8, P=0.1). All patients had adverse events such as weight loss, fever and anorexia. Hospitalization because of decompensation or infection was necessary in three patients. Three patients underwent liver transplantation. A virological response on treatment was achieved in eight patients and sustained in three (33.3%) patients. CONCLUSION: Treatment with standard interferon-alpha2b/ribavirin could be of benefit in patients with advanced liver cirrhosis and thrombocytopenia however, a vigilant monitoring of these high risk patients is mandatory. PMID- 17136335 TI - Urokinase-type plasminogen activator and its inhibitor in thyroid neoplasms: a cytosol study. AB - PURPOSE: Higher levels of urokinase-type plasminogen activator (uPA) and its inhibitor (PAI-1) are linked to the poor prognosis in a variety of malignances. uPA and PAI-1 were expressed in most thyroid carcinomas, as had been measured immunohistochemically. However, no relationship between their expression and clinicopathological parameters were found. Aim of the present study was to investigate the expression and clinical relevance of uPA and PAI-1 in thyroid cancer. PATIENTS AND METHODS: uPA and PAI-1 in paired cytosol samples of thyroid tumor and normal tissue were determined in 23 patients using enzyme-linked immunosorbent assay and correlated to the known prognostic features. RESULTS: Both uPA and PAI-1 concentrations were significantly higher in malignant thyroid tumors (uPA=1.342 +/- 2.944 and PAI-1=17.615 +/- 31.933 ng/mg protein) than in normal tissue (uPA=0.002 +/- 0.009, P=0.011 and PAI-1=2.333 +/- 0.338 ng/mg protein, P=0.001) with positive correlation of the two proteins in the tumors. There were no differences in proteins' levels between benign tumors and normal tissue. Both proteins' concentrations were significantly different among various histological grades (uPA P=0.024 and PAI-1 P=0.017), showing higher values in higher tumor grades (grade I uPA=0.116 +/- 0.247 and PAI-1=4.802 +/- 4.151 ng/mg protein; grade III uPA=8.45 +/- 2.192 and PAI-1=94.65 +/- 59.468 ng/mg protein). The uPA and PAI-1 levels showed significant differences among different histological types of thyroid cancer (uPA P=0.049 and PAI-1=0.017). The lowest values were in adenomas (uPA=0.013 +/- 0.025 and PAI-1=2.785 +/- 1.069 ng/mg protein) and the highest in anaplastic carcinomas (uPA=8.45 +/- 2.192 and PAI 1=94.65 +/- 59.468 ng/mg protein). uPA and PAI-1 were significantly higher in anaplastic vs. well-differentiated cancers (uPA P=0.014 and PAI-1 P=0.026), if extrathyroidal invasion (uPA P=0.019 and PAI-1 P=0.009) or distant metastases (uPA P=0.006 and PAI-1 P=0.003) had been present, and in tumors whose size exceeded 1 cm in diameter (uPA P=0.009 and PAI-1 P=0.035). Only PAI-1, but not uPA was significantly higher in multicentric vs. solitary tumors (P=0.012) and lymph node positive compared to lymph node negative patients (P=0.042). The differences of uPA and PAI-1 did not reach the significant level when patients with well-differentiated tumors below and above 40 years of age had been compared. Survival analysis revealed the significant impact of both uPA and PAI-1 on the Progression-Free Survival (PFS) (38.84 vs. 3.67 months for patients with low and high uPA, respectively, P<0.001; 38.2 vs. 12 months for patients with low and high PAI-1, respectively, P=0.016). CONCLUSIONS: The correlation of high uPA and PAI-1 with the known prognostic factors of poorer outcome and with lower PFS rate in patients with thyroid cancers proved that these proteins could be an additional prognostic parameter. PMID- 17136336 TI - Short course of quinine plus a single dose of sulfadoxine/pyrimethamine for Plasmodium falciparum malaria. AB - BACKGROUND: Quinine remains the treatment of choice in hospitalized malaria cases; however, adverse reactions and the long treatment duration of 7 days often hamper its adequate use. Shortening the treatment by adding sulfadoxine/pyrimethamine may enhance compliance and reduce side effects. We aimed to assess the efficacy of a 3-day course of quinine plus a single dose of sulfadoxine/pyrimethamine for the treatment of non-severe hospitalized malaria cases in Lambarene, Gabon. METHODS: Fifty children aged between 2 and 7 years received quinine dihydrochloride (12 mg/kg every 12 hours for 72 hours), and then a single dose of oral SP (500 mg/25 mg tablet) was given according to weight category. The children were hospitalized for the duration of the treatment and until two consecutive blood smears were negative for malaria parasites. The follow-up period lasted 28 days. RESULTS: Parasites were cleared after 66 hours (SD: 15 hours) and the fever after 46 hours (SD: 24 hours). All patients evaluable by day 28 were negative for malaria parasites (100% efficacy rate, 95% CI: 0.92-1). Only two patients out of 49 had gametocytemia on days 7 and 14. There was no adverse event probably or possibly attributable to the study drugs. CONCLUSIONS: A very high efficacy can be reached using a 3-day course of quinine plus a single dose of sulfadoxine/pyrimethamine for the treatment of non-severe hospitalized malaria cases in our study area. PMID- 17136337 TI - An unusual outcome in a child with hepatosplenic cat-scratch disease. AB - Typical cat-scratch disease (Bartonella henselae infection) in an immunocompetent child is usually associated with a history of scratch, bite or intimate contact with a cat. Most patients develop a non-tender papule in the scratch line after three to ten days. This may persist for only a few days or as long as two to three weeks. During the next two weeks or more, regional lymph nodes that drain the area gradually enlarge and then slowly resolve in more than 10% of patients. The nodes develop overlying erythema and may suppurate. Atypical forms of cat scratch disease occur in a minority of cases and are characterized by ocular or neurological manifestations, hepatosplenic involvement, vertebral osteomyelitis, endocarditis etc. Immunocompromised individuals with B. henselae infection may develop bacillary angiomatosis, bacillary peliosis, and relapsing bacteremia. There have been several reports of hepatosplenic granulomas caused by B. henselae in immunocompetent children. We report a case of a 6-year-old boy with the hepatosplenic form of cat-scratch disease. Despite early diagnosis and long-term antimicrobial treatment, splenectomy could not be avoided. PMID- 17136338 TI - The insulin analog glargine during an unplanned pregnancy. PMID- 17136339 TI - [In memory of Herbert Braunsteiner]. PMID- 17136340 TI - Combined Fourier-transform infrared imaging and desorption electrospray ionization linear ion-trap mass spectrometry for analysis of counterfeit antimalarial tablets. AB - This paper reports use of a combination of Fourier-transform infrared (FTIR) spectroscopic imaging and desorption electrospray ionization linear ion-trap mass spectrometry (DESI MS) for characterization of counterfeit pharmaceutical tablets. The counterfeit artesunate antimalarial tablets were analyzed by both techniques. The results obtained revealed the ability of FTIR imaging in non destructive micro-attenuated total reflection (ATR) mode to detect the distribution of all components in the tablet, the identities of which were confirmed by DESI MS. Chemical images of the tablets were obtained with high spatial resolution. The FTIR spectroscopic imaging method affords inherent chemical specificity with rapid acquisition of data. DESI MS enables high sensitivity detection of trace organic compounds. Combination of these two orthogonal surface-characterization methods has great potential for detection and analysis of counterfeit tablets in the open air and without sample preparation. PMID- 17136341 TI - Determination of UV filters and antimicrobial agents in environmental water samples. AB - Although there is increasing concern about residues from personal care products entering the aquatic environment and their potential to accumulate to levels that pose a health threat to humans and wildlife, we still know little about the extent and magnitude of their presence in the aquatic environment. In this study we describe a procedure for isolation, and subsequent determination, of compounds commonly added to personal care products. The compounds of interest include UV filters with the commercial name Eusolex (homosalate, 4-methylbenzylidenecamphor, benzophenone-3, octocrylene, butylmethoxydibenzoylmethane, ethylhexyl methoxycinnamate) and two common anti-microbial agents, clorophene and triclosan. Water samples were filtered, acidified, and extracted by use of solid-phase extraction. Extracted compounds were then derivatised before analysis by gas chromatography-mass spectroscopy. By use of our method we obtained limits of detection of 13-266 ng L(-1) for UV filters, and 10-186 ng L(-1) for triclosan and clorophene. Recoveries were 82-98% for deionised water and 50-98% for natural water (seawater, pool water, lake water, and river water). Samples collected in Slovenia included seventeen recreational waters (seawater, pool water, lake water, and river water; August 2004) and four wastewaters (January 2005). The most abundant UV filter was benzophenone-3 (11-400 ng L(-1)). Of the two anti microbial agents studied, trace amounts, only, of triclosan were present in the river Kolpa (68 ng L(-1)) and in an hospital effluent (122 ng L(-1)). PMID- 17136342 TI - Femtosecond laser ablation inductively coupled plasma mass spectrometry: achievements and remaining problems. PMID- 17136343 TI - [The cloverleaf plate for osteosythesis of humeral head fractures. Definition of the current position]. AB - In this prospective study we evaluated the functional and radiological results obtained in 62 patients who had been treated for fracture of the humeral head by internal fixation with a cloverleaf plate. They were examined postoperatively, after 14 weeks and after an average follow-up of 75 weeks. Nine patients (14.5%) had dislocated 2-part fractures, 36 had 3-part fractures (58%; additional dislocations of the greater or lesser tubercle), 13 patients (21%) had 4-part fractures and 4 (5.6%) had luxation fractures of the humeral head. Early functional physiotherapy was started on the third day after surgery. The complications observed were: subcutaneous infection (2 cases; 3.2%), haematoma (2 cases; 3.2%), temporal paraesthesia of the axillary nerve (1 case; 1.6%). Only 4 (6.5%) patients suffered from necrosis of the humeral head (partial in 3, total in 1); in 2 cases (3.2%) we switched to a different procedure; in both these patients a humeral head prosthesis was implanted; in both these cases the clinical result was poor because of progressive varus dislocation. To improve mobility we performed arthrolysis in 8 cases (12.9%) and acromioplasty in 10 (16.1%), in addition to removing the plates after fracture consolidation confirmed by X-ray examination. In the present study those of our patients who had been treated with open reduction and internal fixation with a cloverleaf plate achieved average Neer scores of 77+/-13 and average Constant scores of 72.4+/-18, and the rates of complications or revisions were low. "Good" or "very good" results were obtained according to the Constant score in 59% of the treated patients. Even patients with complex 4-part fractures had average Constant scores of 72.7 points ("good"). The accuracy of the refixation of the greater tubercle, sufficiently low fixation of the cloverleaf plate and avoidance of varus position when the humeral head was repositioned were significant parameters influencing the functional outcome in our patients. PMID- 17136344 TI - Water and salinity stress in grapevines: early and late changes in transcript and metabolite profiles. AB - Grapes are grown in semiarid environments, where drought and salinity are common problems. Microarray transcript profiling, quantitative reverse transcription PCR, and metabolite profiling were used to define genes and metabolic pathways in Vitis vinifera cv. Cabernet Sauvignon with shared and divergent responses to a gradually applied and long-term (16 days) water-deficit stress and equivalent salinity stress. In this first-of-a-kind study, distinct differences between water deficit and salinity were revealed. Water deficit caused more rapid and greater inhibition of shoot growth than did salinity at equivalent stem water potentials. One of the earliest responses to water deficit was an increase in the transcript abundance of RuBisCo activase (day 4), but this increase occurred much later in salt-stressed plants (day 12). As water deficit progressed, a greater number of affected transcripts were involved in metabolism, transport, and the biogenesis of cellular components than did salinity. Salinity affected a higher percentage of transcripts involved in transcription, protein synthesis, and protein fate than did water deficit. Metabolite profiling revealed that there were higher concentrations of glucose, malate, and proline in water-deficit treated plants as compared to salinized plants. The metabolite differences were linked to differences in transcript abundance of many genes involved in energy metabolism and nitrogen assimilation, particularly photosynthesis, gluconeogenesis, and photorespiration. Water-deficit-treated plants appear to have a higher demand than salinized plants to adjust osmotically, detoxify free radicals (reactive oxygen species), and cope with photoinhibition. PMID- 17136345 TI - Decreased expression of insulin-like growth factor binding protein 7 in human colorectal carcinoma is related to DNA methylation. AB - PURPOSE: Insulin-like growth factor binding protein 7 (IGFBP-7) is considered a tumor suppressor in various cancers, but its role in colorectal cancer (CRC) is still uncertain. The aims of this study were to analyze the IGFBP-7 expression, and explore the mechanism responsible for the inactivation of IGFBP-7 in CRC. METHODS: mRNA expression was studied by RT-PCR and Northern blot analysis of cultured cells. Methylation status was analyzed by treatment with 5-aza-2' deoxycytidine followed by sequencing of PCR products of sodium bisulfite-treated genomic DNA. IGFBP-7 protein expression was evaluated by immunohistochemistry (IHC) on tissue microarrays. RESULTS: mRNA expression was lost in six out of eight CRC cell lines as compared to normal colon cells. DNA methylation was found in the region of exon 1 and intron 1 of IGFBP-7. In tumor tissue, 107 out of 279 samples showed a negative expression of IGFBP-7 by IHC, which was significantly associated with poor prognosis. The analysis of 37 paired cancerous and normal mucosa samples confirmed the downregulation in the tumors, but revealed variable basal expression levels of IGFBP-7 in normal mucosal samples. CONCLUSIONS: DNA methylation is a mechanism responsible for IGFBP-7 gene silencing providing a target for therapeutic intervention of this tumor suppressor gene. PMID- 17136346 TI - Transcription factor SOX2 up-regulates stomach-specific pepsinogen A gene expression. AB - PURPOSE: Transcription factor SOX2 is expressed in normal gastric mucosae but not in the normal colon. We aimed to clarify the role of SOX2 with reference to pepsinogen expression in the gastrointestinal epithelium. METHODS: We analyzed expression of SOX2 and pepsinogens, differentiation markers of the stomach, in ten gastric cancer (GC) and ten colorectal cancer (CRC) cell lines. The effects of over-expression and down-regulation of SOX2 on pepsinogen expression were also examined. RESULTS: Six GC and five CRC cell lines showed SOX2 expression on RT PCR. Expression of pepsinogen A was detectable in eight GC and seven CRC cell lines, whereas the majority of the cell lines expressed pepsinogen C. Over expression of SOX2 up-regulated expression of pepsinogen A but not that of pepsinogen C in 293T human embryonic kidney cells, and some GC and CRC cell lines. Moreover, pepsinogen A expression was significantly reduced by SOX2 RNA interference in two GC cell lines. CONCLUSION: These data suggest that SOX2 plays an important role in regulation of pepsinogen A, and ectopic expression of SOX2 may be associated with abnormal differentiation of colorectal cancer cells. PMID- 17136347 TI - Psoriasin (S100A7) is significantly up-regulated in human epithelial skin tumours. AB - PURPOSE: Psoriasin (S100A7) has originally been described to be expressed by psoriatic keratinocytes possibly as a result of altered differentiation and inflammation. As psoriasin was found to be overexpressed in human breast and bladder cancer suggesting a role in tumour progression, we investigated the expression of psoriasin in human epithelial skin tumours. METHODS: Realtime reverse transcription-polymerase chain reaction experiments were performed to analyse the mRNA-expression levels of psoriasin together with involucrin as a marker for epithelial differentiation and interleukin-8 (IL-8) as a marker for inflammation in skin biopsy samples from patients with precancerous skin lesions (PSL, n = 6), squamous cell carcinoma (SCC, n = 11), basal cell carcinoma (BCC, n = 17), and healthy controls (n = 10). RESULTS: Unexpectedly, mRNA expression levels for Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) revealed a variation of up to 600-fold in all cDNA-samples under investigation, indicating that GAPDH is not suitable as a housekeeping gene in human skin samples. Psoriasin mRNA expression was significantly up-regulated in samples of PSL, SCC and BCC. In situ hybridisation and immunohistochemical examinations identified psoriasin mRNA and protein expression in the differentiated layers of the epidermis. IL-8 mRNA expression was significantly up-regulated in SCC, however, there was no correlation between elevated levels of psoriasin and the expression of IL-8. CONCLUSION: Similar to the findings in breast and bladder cancer, the up regulation of psoriasin might play a role in the progression of skin cancer. The expression of psoriasin in human skin tumours seems to be independent from differentiation and inflammation. PMID- 17136348 TI - Mutator-like elements identified in melon, Arabidopsis and rice contain ULP1 protease domains. AB - The transposon Mutator was first identified in maize, and is one of the most active mobile elements in plants. The Arabidopsis thaliana genome contains at least 200 Mutator-like elements (MULEs), which contain the Mutator-like transposase gene, and often additional genes. We have detected a novel type of MULEs in melon (CUMULE), which, besides the transposase, contains two ubiquitin like specific protease-like sequences (ULP1). This element is not present in the observed location in some melon cultivars. Multiple copies of this element exist in the Cucumis melo genome, and it has been detected in other Cucurbitaceae species. Analysis of the A. thaliana genome revealed more than 90 CUMULE-like elements, containing one or two Ulp1-like sequences, although no evidence of mobility exists for these elements. We detected various putative transposable elements containing ULP1-like sequences in rice. The discovery of these MULEs in melon and Arabidopsis, and the existence of similar elements in rice and maize, suggest that a proteolytic function may be important for this subset of the MULE transposable elements. PMID- 17136349 TI - DNA elements modulating the KARS12 chromosomal replicator in Kluyveromyces lactis. AB - Eukaryotic chromosomal DNA replication is initiated by a highly conserved set of proteins that interact with cis-acting elements on chromosomes called replicators. Despite the conservation of replication initiation proteins, replicator sequences show little similarity from species to species in the small number of organisms that have been examined. Examination of replicators in other species is likely to reveal common features of replicators. We have examined a Kluyeromyces lactis replicator, KARS12, that functions as origin of DNA replication on plasmids and in the chromosome. It contains a 50-bp region with similarity to two other K. lactis replicators, KARS101 and the pKD1 replication origin. Replacement of the 50-bp sequence with an EcoRI site completely abrogated the ability of KARS12 to support plasmid and chromosomal DNA replication origin activity, demonstrating this sequence is a common feature of K. lactis replicators and is essential for function, possibly as the initiator protein binding site. Additional sequences up to 1 kb in length are required for efficient KARS12 function. Within these sequences are a binding site for a global regulator, Abf1p, and a region of bent DNA, both of which contribute to the activity of KARS12. These elements may facilitate protein binding, protein/protein interaction and/or nucleosome positioning as has been proposed for other eukaryotic origins of DNA replication. PMID- 17136350 TI - Positive polarotaxis in a mayfly that never leaves the water surface: polarotactic water detection in Palingenia longicauda (Ephemeroptera). AB - Tisza mayflies, Palingenia longicauda (Olivier 1791), swarm exclusively over the river Tisza (from which the name of the mayfly was derived). This river is bordered by a high vertical wall of trees and bushes, which hinder P. longicauda to move away horizontally from the water. During swarming, Tisza mayflies fly immediately above the river in such a way that their cerci touch the water frequently or sweep its surface. This continuous close connection with water and the vertical wall of the shore and riparian vegetation result in that Tisza mayflies never leave the water surface; consequently, they need not search for water. Several Ephemeroptera species move away far from water and return to it guided by the horizontal polarization of water-reflected light. To reveal whether also P. longicauda is or is not polarotactic, we performed a field experiment during the very short swarming period of Tisza mayflies. We show here that also P. longicauda has positive polarotaxis, which, however, can be observed only under unnatural conditions, when the animals are displaced from the water and then released above artificial test surfaces. P. longicauda is the first species in which polarotactic water detection is demonstrated albeit it never leaves the water surface, and thus, a polarotactic water detection seems unnecessary for it. The polarotactic behaviour of Tisza mayflies explains the earlier observation that these insects swarm above wet asphalt roads running next to river Tisza. PMID- 17136351 TI - Evaluation of maternal measles antibody titers and response to the measles vaccination at the age of 9 months in preterm infants. PMID- 17136352 TI - IgG1 deficiency and high IgA level with juvenile idiopathic arthritis. PMID- 17136353 TI - The relationship between immune thrombocytopenic purpura and Helicobacter pylori infection in children: where is the truth? PMID- 17136354 TI - Serotype distribution, clonality and antimicrobial resistance of invasive pneumococcal isolates in a central Italian region: implications for vaccine strategies. PMID- 17136355 TI - Determination of ANA specificity using multiplexed fluorescent microsphere immunoassay in patients with ANA positivity at high titres after infliximab treatment: preliminary results. AB - To evaluate ANA specificity using the fully automated multiplexed fluorescent microsphere immunoassay in patients affected either by rheumatoid arthritis or ankylosing spondylitis who developed strong positivity for ANA as assessed by indirect immunofluorescent method on HEp-2 cells during infliximab treatment. Three men affected by ankylosing spondylitis and 12 women affected by rheumatoid arthritis who developed ANA positivity at high titres during infliximab treatment underwent the identification of ANA specificity by multiplexed fluorescent microsphere immunoassay; moreover anti-DNA and anti-ENA antibodies were tested by indirect immunofluorescence and ELISA method, respectively. In 4 out of 15 cases, the determination of ANA reactivity by multiplexed fluorescent microsphere immunoassay was also performed on the serum collected before infliximab administration. One patient affected by rheumatoid arthritis showed multiple ANA reactivities against SS-A, SS-B, RNP, Sm, Jo-1 and histones; one patient affected by ankylosing spondylitis resulted positive for the same autoantibodies, except for anti-Sm antibody. Moreover, two patients, one with rheumatoid arthritis and one with ankylosing spondylitis, showed single antibody specificity to SS-B and RNP, respectively. The remaining 11 cases did not show any positivity. Instead, all the patients resulted negative for anti-ENA antibodies by the ELISA method. In the four cases tested for ANA specificity by multiplexed fluorescent microsphere immunoassay before and after infliximab administration no difference was found. The search for anti-DNA antibody always resulted negative by both the traditional immunofluorescent assay and the novel technique. The use of multiplexed fluorescent microsphere immunoassay in patients treated with infliximab with ANA positivity at high titres allowed to find some ANA specificities which were not revealed by ELISA method. Nevertheless, the majority of patients resulted negative in spite of ANA positivity at high titres; the molecular target of ANA which develop after infliximab administration still remains to be identified. PMID- 17136356 TI - 1H-NMR metabolic profiling of human neonatal urine. AB - OBJECT: The measurement of different urine components and their changes over time may provide comprehensive and early information about perinatal metabolic processes and physiological changes. We hypothesized that (1) H-NMR-spectroscopy generating a complex spectral profile without pre-selection of urinary metabolites could identify metabolites determining the neonatal physiological status and discriminating between different metabolic states. MATERIALS AND METHODS: We studied spot urine of three groups of neonates (healthy term-born, term-born with non-specific bacterial infections, and preterm neonates) for the first 6 days of life using (1) H-NMR-spectroscopy. In the group of healthy neonates metabolites changing were identified and their excretion patterns compared between groups. RESULTS: Six metabolites indicating physiological changes were identified: N-methylnicotinamide (NAD (+)-pathway), formate, hippurate, betaine (kidney development), taurine (neuronal development), and bile acids (hepatic clearance). While the dynamic changes over the first 6 days were the same for all metabolites in both groups of term-born neonates, the excretion of N-methylnicotinamide and taurine was significantly higher in preterm neonates compared to healthy term neonates and neonates with bacterial infections from the third day after birth (P < 0.05). CONCLUSION: Urine analysis using (1) H-NMR spectroscopy could identify markers for perinatal metabolic changes. Further studies have to clarify if the proposed physiological interpretation will correlate with long-term physiological development. PMID- 17136357 TI - Intravitreal bevacizumab (Avastin) associated with the regression of subretinal neovascularization in idiopathic juxtafoveolar retinal telangiectasis. AB - PURPOSE: To describe the clinical, angiographic, and optical coherence tomography findings of a patient with subretinal neovascularization (SRN) in idiopathic juxtafoveolar retinal telangiectasis (IJRT) treated with intravitreal bevacizumab injection. METHODS: In the setting of a tertiary referral center, a patient with bilateral acquired IJRT, complicated with juxtafoveal SRN in one eye, was treated with a single intravitreal injection of 1.5 mg of bevacizumab and then prospectively followed for 24 weeks. RESULTS: Vision improved from 20/40(-1) to 20/20(-2) by 24 weeks of follow-up. Fluorescein angiography demonstrated the absence of leakage from a previously active SRN. Optical coherence tomography revealed gradual neovascular lesion contraction and the resolution of subretinal fluid over the 24-week follow-up period. No obvious adverse events were noted. CONCLUSIONS: In the short-term, intravitreal bevacizumab treatment resulted in partial restoration of the macular architecture and vision improvement by means of neovascular lesion regression and the resolution of associated serohemorrhagic complications. PMID- 17136358 TI - Contributions of matrix metalloproteinases toward Meckel's cartilage resorption in mice: immunohistochemical studies, including comparisons with developing endochondral bones. AB - The middle portion of Meckel's cartilage (one of four portions that disappear with unique fate) degrades via hypertrophy and the cell death of chondrocytes and via the resorption of cartilage by chondroclasts. We have examined the immunolocalization of matrix metalloproteinase-2 (MMP-2), MMP-9, MMP-13, and MMP 14 (members of the MMP activation cascade) and galectin-3 (an endogenous substrate for MMP-9 and an anti-apoptotic factor) during resorption of Meckel's cartilage in embryonic mice and have compared the results with those of developing endochondral bones in hind limbs. MMP immunoreactivity, except for MMP 2, is present in nearly all chondrocytes in the middle portion of Meckel's cartilage. On embryonic day 15 (E15), faint MMP-2-immunoreactive and intense MMP 13-immunoreactive signals occur in the periosteal bone matrix deposited by periosteal osteoblasts on the lateral surface, whereas MMP-9 and MMP-14 are immunolocalized in the peripheral chondrocytes of Meckel's cartilage. The activation cascade of MMPs by face-to-face cross-talk between cells may thus contribute to the initiation of Meckel's cartilage degradation. On E16, immunopositive signaling for MMP-13 is detectable in the ruffled border of chondroclasts at the resorption front, whereas immunostaining for galectin-3 is present at all stages of chondrocyte differentiation, especially in hypertrophic chondrocytes adjacent to chondroclasts. Galectin-3-positive hypertrophic chondrocytes may therefore coordinate the resorption of calcified cartilage through cell-to-cell contact with chondroclasts. In metatarsal specimens from E16, MMPs are detected in osteoblasts, young osteocytes, and the bone matrix of the periosteal envelope, whereas galectin-3 immunoreactivity is intense in young periosteal osteocytes. In addition, intense MMP-9 and MMP-14 immunostaining has been preferentially found in pre-hypertrophic chondrocytes, although galectin-3 immunoreactivity markedly decreases in hypertrophic chondrocytes. These results indicate that the degradation of Meckel's cartilage involves an activation cascade of MMPs that differs from that in endochondral bone formation. PMID- 17136359 TI - Trunk biomechanical models based on equilibrium at a single-level violate equilibrium at other levels. AB - Accurate estimation of muscle forces in various occupational tasks is critical for a reliable evaluation of spinal loads and subsequent assessment of risk of injury and management of back disorders. The majority of biomechanical models of multi-segmental spine estimate muscle forces and spinal loads based on the balance of net moments at a single level with no consideration for the equilibrium at remaining levels. This work aimed to quantify the extent of equilibrium violation and alterations in estimations when such models are performed at different levels. Results are compared with those of kinematics driven model that satisfies equilibrium at all levels and EMG data. Regardless of the method used (optimization or EMG-assisted), single-level free body diagram models yielded estimations that substantially altered depending on the level considered (i.e., level dependency). Equilibrium of net moment was also grossly violated at remaining levels with the error increasing in more demanding tasks. These models may, however, be used to estimate spinal compression forces. PMID- 17136360 TI - Diameter of the vestibular aqueduct. PMID- 17136361 TI - Arthropathy of neonatal onset multisystem inflammatory disease (NOMID/CINCA). AB - BACKGROUND: Neonatal onset multisystem inflammatory disease (NOMID), an autoinflammatory disease, is characterized by fever, chronic urticarial rash, CNS manifestations, and arthropathy. Approximately 50% of patients with NOMID have de novo missense mutations in CIAS1, which is associated with modulation of the IL 1b and apoptotic pathways. Approximately 60% of NOMID patients have prominent arthropathy, most commonly involving the knees, the cause of which remains poorly understood. OBJECTIVE: To more fully describe the findings of NOMID arthropathy on MRI and radiography and to provide a better understanding of the origin of the bony lesions. MATERIALS AND METHODS: We imaged 20 patients with NOMID to further investigate NOMID-associated bony lesions. RESULTS: Bony abnormalities were seen in the knees of 11/20 patients. The knee findings included enlarged, deformed femora and patellae in all and tibiae in the majority, without evidence of synovitis. Some patients had other joint involvement. Most had short stature and valgus or varus knee deformities. No association was noted between bony abnormalities and CIAS1 mutations. The abnormalities appeared to be the result of a mass-producing process. The resulting heterogeneously calcified masses appeared to originate in the physis and deformed the adjacent metaphysis and epiphysis. CONCLUSION: These findings suggest that the arthropathy of NOMID is the result of abnormal endochondral bone growth. Further investigation is needed to determine whether this deformity is triggered by inflammation early in development or by CIAS1 mutations causing abnormal chondrocyte apoptosis. PMID- 17136362 TI - Water imaging (hydrography) in the fetus: the value of a heavily T2-weighted sequence. AB - BACKGROUND: Since the development of fast imaging sequences, MR has proved to be a helpful tool in the evaluation of fetal pathology. Because of the high water content of fetal tissues and pathology, hydrography imaging (MR fetography) can provide additional diagnostic information. OBJECTIVE: To demonstrate the benefit of MR fetography in fetal imaging. MATERIALS AND METHODS: From 2004 to 2005, 126 fetal MR examinations were performed for evaluation of an abnormality depicted on an antenatal sonogram. Single-shot fast spin-echo MR imaging and MR fetography were performed through the area of fetal pathology. The two studies were retrospectively compared. RESULTS: The primary diagnosis was not changed with the addition of MR fetography. New findings, particularly in the kidneys and spine, were identified in 9% of the patients. When fetal pathology was of high water content (80% patients), the MR fetography imaging increased diagnostic confidence. In 11% of the patients, those with cardiovascular or low water pathology, the MR fetography was not beneficial. CONCLUSION: The mainstay of fetal imaging is currently the HASTE and SSFSE sequences. However, MR fetography is an excellent adjunct that highlights fetal pathology by reinforcing the diagnosis, identifying additional findings, and providing high-contrast high resolution images that are helpful when counseling clinicians and patients. PMID- 17136366 TI - Leaching and microbial treatment of a soil contaminated by sulphide ore ashes and aromatic hydrocarbons. AB - Contaminated soil from a historical industrial site and containing sulfide ore ashes and aromatic hydrocarbons underwent sequential leaching by 0.5 M citrate and microbial treatments. Heavy metals leaching was with the following efficiency scale: Cu (58.7%) > Pb (55.1%) > Zn (44.5%) > Cd (42.9%) > Cr (26.4%) > Ni (17.7%) > Co (14.0%) > As (12.4%) > Fe (5.3%) > Hg (1.1%) and was accompanied by concomitant removal of organic contaminants (about 13%). Leached metals were concentrated into an iron gel, produced during ferric citrate fermentation by the metal-resistant strain BAS-10 of Klebsiella oxytoca. Concomitantly, the acidic leached soil was bioaugmented with Allescheriella sp. DABAC 1, Stachybotrys sp. DABAC 3, Phlebia sp. DABAC 9, Pleurotus pulmonarius CBS 664.97, and Botryosphaeria rhodina DABAC P82. B. rhodina was most effective, leading to a significant depletion of the most abundant contaminants, including 7-H benz[DE]anthracene-7-one, 9,10-anthracene dione and dichloroaniline isomers, and to a marked detoxification as assessed by the mortality test with the Collembola Folsomia candida Willem. The overall degradation activities of B. rhodina and P. pulmonarius appeared to be significantly enhanced by the preliminary metal removal. PMID- 17136367 TI - Probiotic properties of Lactobacillus and Bifidobacterium strains isolated from porcine gastrointestinal tract. AB - One strain of Lactobacillus salivarius, two strains of Lactobacillus reuteri and Lactobacillus amylovorus, and two strains of Bifidobacterium thermacidophilum with antagonistic effect against Clostridium perfringens were isolated from porcine gastrointestinal tract. Isolates were assayed for their ability to survive in synthetic gastric juice at pH 2.5 and were examined for their ability to grow on agar plate containing porcine bile extract. There was a large variation in the survival of the isolates in gastric juice and growth in the medium containing 0.3% (w/v) bile. L. salivarius G11 and L. amylovorus S6 adhered to the HT-29 epithelial cell line. Cell-free supernatant of L. amylovorus S6 showed higher antagonistic activity as effective as the antibiotics such as neomycin, chlortetracycline, and oxytetracycline against bacterial pathogens including C. perfringens, Salmonella typhimurium, Staphylococcus aureus, Vibrio cholerae, Edwardsiella tarda, and Aeromonas salmonicida subsp. salmonicida. PMID- 17136368 TI - Remaining acetamide in acetonitrile degradation using nitrile hydratase- and amidase-producing microorganisms. AB - The tandem conversion process involving nitrile hydratase- and amidase-producing microorganisms has potential for use in the treatment of acetonitrile-containing wastes. In that process, the acetamide hydrolysis step catalyzed by amidase is very slow compared with the acetonitrile hydration step catalyzed by nitrile hydratase, and a small amount of acetamide remains in the resulting solution. This study aimed to improve the efficiency of the acetamide hydrolysis step. An amidase-producing microorganism, Rhodococcus sp. S13-4, was newly obtained, whose use enabled rapid acetamide degradation. Though residual acetamide was still detected, it was successfully reduced by the addition of cation/anion mixed ion exchange resin or calcium hydroxide after the acetamide hydrolysis reaction using Rhodococcus sp. S13-4 cells. This result implies that acetamide hydrolysis and acetamide formation are in equilibrium. The incubation of Rhodococcus sp. S13-4 cells with high concentrations of ammonium acetate produced acetamide. The purified amidase from Rhodococcus sp. S13-4 revealed the acetamide formation activity (specific activity of 30.6 U/mg protein). This suggests that the amidase catalyzed amide formation may cause the remaining of acetamide in the acetonitrile conversion process. PMID- 17136369 TI - Phytohormone production by three strains of Bradyrhizobium japonicum and possible physiological and technological implications. AB - The aim of this work was to evaluate phytohormone biosynthesis, siderophores production, and phosphate solubilization in three strains (E109, USDA110, and SEMIA5080) of Bradyrhizobium japonicum, most commonly used for inoculation of soybean and nonlegumes in USA, Canada, and South America. Siderophore production and phosphate solubilization were evaluated in selective culture conditions, which had negative results. Indole-3-acetic acid (IAA), gibberellic acid (GA(3)), and abscisic acid (ABA) production were analyzed by gas chromatography-mass spectrometry (GC-MS). Ethylene and zeatin biosynthesis were determined by GS flame ionization detection and high-performance liquid chromatography (HPLC-UV), respectively. IAA, zeatin, and GA(3) were found in all three strains; however, their levels were significantly higher (p < 0.01) in SEMIA5080 (3.8 microg ml( 1)), USDA110 (2.5 microg ml(-1)), and E109 (0.87 microg ml(-1)), respectively. ABA biosynthesis was detected only in USDA110 (0.019 microg ml(-1)). Ethylene was found in all three strains, with highest production rate (18.1 ng ml(-1) h(-1)) in E109 cultured in yeast extract mannitol medium plus L-methionine. This is the first report of IAA, GA(3), zeatin, ethylene, and ABA production by B. japonicum in pure cultures, using quantitative physicochemical methodology. The three strains have differential capability to produce the five major phytohormones and this fact may have an important technological implication for inoculant formulation. PMID- 17136370 TI - Effect of postpartum counseling on postpartum contraceptive use. AB - OBJECTIVE: The aim of this study was to evaluate the effect of postpartum counseling on postpartum contraceptive use. METHODS: One hundred and forty-three women who delivered between 1 January 2004 and 31 September 2004 and counseled about postpartum contraception were included in the study. The participants were interviewed by telephone. Age, gravidity, parity, and mode of delivery of the participants were recorded. Their method of contraception before pregnancy, their decision on the contraceptive method after counseling and the method actually used were asked. RESULTS: Just after postpartum counseling, 47 women (32.9%) decided to use the intrauterine device (IUD), 23 (16.1%) condoms, 16 (11.2%) progestin injections, 7 (4.9%) oral contraceptives, and 7 (4.9%) coitus interruptus for contraception. Thirty-six women (25.2%) did not decide on any method of use. At the time of the telephone interview the actual method used was learned. Fifty-one women (35.7%) were using coitus interruptus, 45 women (31.5%) condoms, and 14 (9.8%) the IUD. Sixteen women (11.2%) were reported as not using any methods. CONCLUSION: In spite of postpartum counseling, a high majority of the women appeared to use traditional and less effective contraceptive methods. PMID- 17136371 TI - Mapping and characterization of FLC homologs and QTL analysis of flowering time in Brassica oleracea. AB - The FLC gene product is an inhibitor of flowering in Arabidopsis. FLC homologs in Brassica species are thought to control vernalization. We cloned four FLC homologs (BoFLCs) from Brassica oleracea. Three of these, BoFLC1, BoFLC3 and BoFLC5, have been previously characterized. The fourth novel sequence displayed 98% sequence homology to the previously identified gene BoFLC4, but also showed 91% homology to BrFLC2 from Brassica rapa. Phylogenetic analysis showed that this clone belongs to the FLC2 clade. Therefore, we designated this gene BoFLC2. Based on the segregation of RFLP, SRAP, CAPS, SSR and AFLP loci, a detailed linkage map of B. oleracea was constructed in the F(2) progeny obtained from a cross of B. oleracea cv. Green Comet (broccoli; non-vernalization type) and B. oleracea cv. Reiho (cabbage; vernalization type), which covered 540 cM, 9 major linkage groups. Six quantitative trait loci (QTL) controlling flowering time were detected. BoFLC1, BoFLC3 and BoFLC5 were not linked to the QTLs controlling flowering time. However, the largest QTL effect was located in the region where BoFLC2 was mapped. Genotyping of F(2 )plants at the BoFLC2 locus showed that most of the early flowering plants were homozygotes of BoFLC-GC, whereas most of the late- and non-flowering plants were homozygotes of BoFLC-Rei. The BoFLC2 homologs present in plants of the non-vernalization type were non-functional, due to a frameshift in exon 4. Moreover, duplications and deletions of BoFLC2 were detected in broccoli and a rapid cycling line, respectively. These results suggest that BoFLC2 contributes to the control of flowering time in B. oleracea. PMID- 17136372 TI - Genetic diversity analysis of traditional and improved Indonesian rice (Oryza sativa L.) germplasm using microsatellite markers. AB - The archipelago of Indonesia has a long history of rice production across a broad range of rice-growing environments resulting in a diverse array of local Indonesian rice varieties. Although some have been incorporated into modern breeding programs, the vast majority of these landraces remain untapped. To better understand this rich source of genetic diversity we have characterized 330 rice accessions, including 246 Indonesian landraces and 63 Indonesian improved cultivars, using 30 fluorescently-labeled microsatellite markers. The landraces were selected across 21 provinces and include representatives of the classical subpopulations of cere, bulu, and gundil rices. A total of 394 alleles were detected at the 30 simple sequence repeat loci, with an average number of 13 alleles per locus across all accessions, and an average polymorphism information content value of 0.66. Genetic diversity analysis characterized the Indonesian landraces as 68% indica and 32% tropical japonica, with an indica gene diversity of 0.53 and a tropical japonica gene diversity of 0.56, and a Fst of 0.38 between the two groups. All of the improved varieties sampled were indica, and had an average gene diversity of 0.46. A set of high quality Indonesian varieties, including Rojolele, formed a separate cluster within the tropical japonicas. This germplasm presents a valuable source of diversity for future breeding and association mapping efforts. PMID- 17136374 TI - [Controlled hypothermia. More than just cooling?]. PMID- 17136373 TI - Root-knot nematode (Meloidogyne spp.) Me resistance genes in pepper (Capsicum annuum L.) are clustered on the P9 chromosome. AB - The root-knot nematode (Meloidogyne spp.) is a major plant pathogen, affecting several solanaceous crops worldwide. In Capsicum annuum, resistance to this pathogen is controlled by several independent dominant genes--the Me genes. Six Me genes have previously been shown to be stable at high temperature in three highly resistant and genetically distant accessions: PI 322719, PI 201234, and CM334 (Criollo de Morelos 334). Some genes (Me4, Mech1, and Mech2) are specific to certain Meloidogyne species or populations, whereas others (Me1, Me3, and Me7) are effective against a wide range of Meloidogyne species, including M. arenaria, M. javanica, and M. incognita, the most common species in Mediterranean and tropical areas. These genes direct different response patterns in root cells depending on the pepper line and nematode species. Allelism tests and fine mapping using the BSA-AFLP approach showed these genes to be different but linked, with a recombination frequency of 0.02-0.18. Three of the PCR-based markers identified in several genetic backgrounds were common to the six Me genes. Comparative mapping with CarthaGene software indicated that these six genes clustered in a single genomic region within a 28 cM interval. Four markers were used to anchor this cluster on the P9 chromosome on an intraspecific reference map for peppers. Other disease resistance factors have earlier been mapped in the vicinity of this cluster. This genomic area is colinear to chromosome T12 of tomato and chromosome XII of potato. Four other nematode resistance genes have earlier been identified in this area, suggesting that these nematode resistance genes are located in orthologous genomic regions in Solanaceae. PMID- 17136375 TI - [Perioperative thermal management]. AB - Perioperative hypothermia can influence clinical outcome negatively. It triples the incidence of adverse myocardial outcomes, significantly increases perioperative blood loss, significantly augments allogenic transfusion requirements, and increases the incidence of surgical wound infections. The major causes are redistribution of heat from the core of the body to the peripheral tissues and a negative heat balance. Adequate thermal management includes preoperative and intraoperative measures. Preoperative measures, e.g., prewarming, enhance heat content of the peripheral tissues, thereby reducing redistribution of heat from the core to the peripheral tissues after induction of anesthesia. Intraoperative measures are active skin surface warming of a large body surface area with conductive or convective warming systems. Intravenous fluids should be warmed when large volumes of more than 500-1000 ml/h are required. The body surfaces that cannot be actively warmed should be insulated. Airway humidification and conductive warming of the back are less efficient. PMID- 17136376 TI - Acute colonic diverticulitis in a community-based hospital: CT evaluation in 138 patients. AB - The purpose of this research is to retrospectively analyze computed tomography (CT) findings of patients with acute colonic diverticulitis presented to a community-based hospital. During a 1-year period from May 2004 to April 2005, CT scans of 138 patients [mean age 62.3 years (SD 14.9), range 30-100 years] with clinical diagnosis of diverticulitis were analyzed. Of the 138 patients, 136 CT scans were performed with oral and intravenous contrast administration except in two patients who received only oral contrast. Twenty-eight patients (28/138, 20.3%) had diverticulosis, 77 (77/138, 55.8%) had uncomplicated diverticulitis, and 33 (33/138, 23.9%) had complications. Left-sided diverticulitis predominated (137/138, 99.3%). Colonic diverticula were identified in almost every patient (136/138, 98.6%); the majority had moderate colonic wall thickening (82/110, 74.5%) and moderate pericolonic inflammation (65/110, 59.1%). Isolated extraluminal air bubbles (19/33, 57.6%) were the most commonly seen complication, followed by abscess (15/33, 45.5%). Bowel obstruction (2/33, 6.1%) and pylephlebitis (1/33, 3.0%) were less commonly seen but were observed as significant abnormalities. Complicated diverticulitis was less prevalent in this community hospital setting, in contrast with referral medical centers. Common CT findings of diverticulitis included presence of diverticula, moderate wall thickening, and pericolonic inflammation. Isolated air bubbles were the most commonly seen complication followed by abscess. PMID- 17136377 TI - Diaphragmatic rupture: a frequently missed injury in blunt thoracoabdominal trauma patients. AB - In the US and Western Europe, trauma is the fourth most common cause of death and the leading cause of death in the population less than 45 years of age [Mullinix and Foley, J Comput Assist Tomogr 28(Suppl 1):S20-S27, 2004]. Diaphragmatic injuries occur in 0.8 to 8% of patients after blunt trauma (Gray H, The muscles of the thorax. Anatomy of the human body. Lea & Febiger, Philadelphia, 1918) and may be a predictor of severity of injury in the blunt trauma patient [Worthy et al., Radiology 194(3):885-888, 1995]. The clinical diagnosis of diaphragmatic rupture (DR) is difficult and is missed in anywhere from 7 to 66% of patients [Cantwell, Radiology 238(2):752-753, 2006]. The accurate diagnosis and prognosis of this pathology depend on a complete knowledge of the clinical and radiological presentation. Computed tomography is the imaging modality of choice in the assessment of patients with clinical or radiographic findings suggestive of DR. PMID- 17136378 TI - MDCT for suspected pulmonary embolism: multi-institutional survey of 16-MDCT data acquisition protocols. AB - The purpose of this study was to determine the extent to which a consensus exists on multidetector row computed tomography (MDCT) protocol parameters for suspected pulmonary embolism (PE). In August of 2004, a questionnaire addressing a number of body MDCT protocols was mailed to 99 fellows of the Society of Computed Body Tomography, representing a total of 46 institutions. In May 2005, this was followed up with a second mailing. The survey requested details pertaining to protocols for the most advanced MDCT scanner in the department. The overall survey response rate of 37% (17/46) yielded 15 protocols for 16-MDCT imaging of suspected PE. This data was tabulated and revealed a consensus for the use of bolus tracking, rapid contrast infusion, caudo-cranial scanning, the narrowest detector row collimation, and thin (<2 mm) reconstruction sections. However, contrast infusion timing, contrast concentration, and implementation of radiation dose modulation were variable. This compilation of protocols reflects recently published studies advocating the use of narrow acquisition collimation and reconstruction sections for MDCT of suspected PE. Future studies are necessary to elucidate the optimal intravenous contrast infusion parameters and further assess the efficacy of reduced radiation dose protocols. PMID- 17136379 TI - Magnetic resonance imaging findings in patients with peroneal tendinopathy and peroneal tenosynovitis. AB - OBJECTIVE: To compare the magnetic resonance (MR) imaging findings of a group of patients with clinically diagnosed peroneal tendonopathy and peroneal tenosynovitis with the MR imaging findings of a control group of patients with no clinical evidence of peroneal tendon disorder. SUBJECTS AND METHODS: The MR examinations of 24 patients with symptomatic peroneal tendinopathy or peroneal tenosynovitis and 70 patients with no clinical evidence of peroneal tendon disorder were retrospectively reviewed to determine the presence or absence of four MR imaging findings: 1) predominantly or uniform intermediate signal intensity within the peroneal tendons on one or more axial proton density weighted images, 2) predominantly or uniform intermediate signal intensity within the peroneal tendons on three consecutive axial proton density-weighted images, 3) intermediate T2 signal intensity within the peroneal tendons, and 4) circumferential fluid within the common peroneal tendon sheath greater than 3 mm in maximal width. The sensitivity and specificity of these MR imaging findings for determining the presence or absence or symptomatic peroneal tendinopathy or peroneal tenosynovitis were calculated. RESULTS: The sensitivity of MR imaging findings 1, 2, 3, and 4 for determining the presence of peroneal tendinopathy or peroneal tenosynovitis were 92%, 92%, 50%, and 17% respectively. The specificity of MR imaging findings 1, 2, 3, and 4 for determining the absence of peroneal tendinopathy or peroneal tenosynovitis were 57%, 79%, 93%, and 100% respectively. CONCLUSION: The presence of predominantly or uniform intermediate signal intensity within the peroneal tendons on three consecutive axial proton density weighted images is a highly sensitive and moderately specific indicator of symptomatic peroneal tendinopathy. The presence of intermediate T2 signal within the peroneal tendons, and the presence of circumferential fluid within the peroneal tendon sheath greater than 3 mm in maximal width, are highly specific indicators of peroneal tendinopathy and peroneal tenosynovitis respectively. PMID- 17136380 TI - Generation and reshaping of sequences in neural systems. AB - The generation of informational sequences and their reorganization or reshaping is one of the most intriguing subjects for both neuroscience and the theory of autonomous intelligent systems. In spite of the diversity of sequential activities of sensory, motor, and cognitive neural systems, they have many similarities from the dynamical point of view. In this review we discus the ideas, models, and mathematical image of sequence generation and reshaping on different levels of the neural hierarchy, i.e., the role of a sensory network dynamics in the generation of a motor program (hunting swimming of marine mollusk Clione), olfactory dynamical coding, and sequential learning and decision making. Analysis of these phenomena is based on the winnerless competition principle. The considered models can be a basis for the design of biologically inspired autonomous intelligent systems. PMID- 17136381 TI - Functional expression of Escherichia coli fhuA gene in Rhizobium spp. of Cajanus cajan provides growth advantage in presence of Fe3+: ferrichrome as iron source. AB - Cajanus cajan rhizobial isolates were found to be unable to utilize iron bound to ferrichrome, desferrioxamine B or rhodotorulic acid, all being hydroxamate type siderophores. A broad host range expression vector containing the Escherichia coli fhuA gene, encoding the outer membrane receptor for Fe-ferrichrome, was constructed. The plasmid construct (pGR1), designed to express fhuA under the lac promoter of E. coli, complemented E. coli MB97 DeltafhuA mutant for ferri ferrichrome utilization and also allowed Rhizobium spp. ST1 and Rhizobium spp. IC3123 to grow using iron bound to ferrichrome. Sensitivity to the antibiotic albomycin, transported via the FhuA receptor, was found in case of MB97 as well as rhizobial transformants harboring pGR1. The rhizobial transformants expressing fhuA showed growth stimulation when co-inoculated with Ustilago maydis, a fungal species known to produce ferrichrome under iron starved conditions. Growth stimulation was also observed in the presence of externally supplied ferrichrome. The significance of these findings in terms of the potential for improving the survivability of rhizobial bioinoculant strains in natural soils is discussed. PMID- 17136382 TI - N-acyl homoserine lactones are degraded via an amidolytic activity in Comamonas sp. strain D1. AB - Comamonas strain D1 enzymatically inactivates quorum-sensing (QS) signal molecules of the N-acyl homoserine lactone (N-AHSL) family, and exhibits the broadest inactivation range of known bacteria. It degrades N-AHSL with acyl-side chains ranging from 4 to 16 carbons, with or without 3-oxo or 3-hydroxy substitutions. N-AHSL degradation yields HSL but not N-acyl homoserine: strain D1 therefore harbors an amidohydrolase activity. Strain D1 is the fifth bacterium species in which an N-AHSL amidohydrolase is described. Consistent with its N AHSL degradation ability, strain D1 efficiently quenches various QS-dependent functions in other bacteria, such as violacein production by Chromobacterium violaceum and pathogenicity and antibiotic production in Pectobacterium. PMID- 17136383 TI - Locomotor response to light and surface angle in three species of desert fleas. AB - We studied the relocation of newly emerged and fed individuals of three species of desert fleas (Xenopsylla conformis, Xenopsylla ramesis, and Parapulex chephrenis) in response to light and surface angle. We observed flea movements inside of either horizontal or tilted cardboard tubes with different light regime at their ends. Proportion of relocating X. conformis and X. ramesis was significantly higher than that of P. chephrenis. In this species only, adult individuals relocated more frequently than newly emerged individuals, and females relocated more frequently than males. In general, the majority of fleas moved toward light independently of its position in relation to the surface angle. Fleas moved toward light even if it was positioned at the lower end of a tube. When both ends of a tube were darkened, newly emerged Xenopsylla moved randomly toward the upper or lower end of a tube, whereas newly emerged P. chephrenis moved mainly toward the upper end of a tube. Adult P. chephrenis and X. conformis also moved mainly toward the upper end of a tube, whereas adult X. ramesis moved mainly toward the lower end. When both ends of a tube were lighted up, newly emerged females of all species, as well as newly emerged female X. ramesis, randomly relocated toward the upper or lower end of a tube. In contrast, newly emerged males and adults of both sexes of P. chephrenis and X. conformis as well as adult female X. ramesis moved mainly toward the upper end of a tube, whereas adult male X. ramesis moved mainly down. Results of this study suggest that light is a more important abiotic signal for flea orientation than surface angle, and there are species-specific differences in flea responses to light and angle stimuli. These differences are related to spatial ecology and behavior of fleas' main hosts as well as to fleas' environmental preferences. PMID- 17136384 TI - Acanthamoeba as a temporal vehicle of Cryptosporidium. AB - The capacity of Acanthamoeba to predate Cryptosporidium oocysts was demonstrated. A maximum of six oocysts per Acanthamoeba trophozoite were detected, and a slow elimination of the internalized oocysts to the surrounding culture medium was observed. Free-living amoebae may act as carriers of Cryptosporidium oocysts and, thus, may play an important role in the transmission of cryptosporidiosis. PMID- 17136385 TI - Serine protease activity demonstrated in the larval stage of the pentastomid Linguatula serrata. AB - Linguatula serrata is a cosmopolitan parasite whose intermediate hosts are cattle, goats, sheep, and other ruminants. The adult form is found in the nasal airways, frontal sinuses, and tympanic cavity of canines and felines, and it produces hemorrhages and breathing difficulties. To elucidate if L. serrata produces enzymes that are capable of degrading tissues from the intermediate host, proteolytic activities in larval products were studied. Using the zymography technique, one major protease was detected in parasite in vitro released products with an approximate molecular weight of 75 kDa. This enzyme was inhibited with phenylmethylsulfonyl fluoride, suggesting that it is a serine protease, which was also shown to degrade type I collagen. The serine protease exhibited maximal activity at alkaline pH and temperatures varying from 37 to 45 degrees C. To gather evidence about the physiological roles of the enzyme, further biochemical and functional studies are suggested. PMID- 17136386 TI - High levels of human chitotriosidase hinder the formation of peritrophic membrane in anopheline vectors. AB - In the Anopheles midgut, Plasmodium falciparum produces a specific chitinase able to penetrate the blood meal surrounding the chitin-containing peritrophic membrane (PM). High levels of an analogous chitinase, chitotriosidase (CHIT), may be found in human blood, being the markers of macrophage activation. To verify the hypothesis that CHIT present in malaria patient blood could help parasite to overcome PM, we carried out a bioassay by feeding Anopheles stephensi females on an artificial apparatus that contained human blood from four different sources and with different chitinase concentrations: (1) healthy donors, as negative controls; (2) patients with malaria; (3) patients with Gaucher disease; and (4) whole blood enriched with commercial P. falciparum chitinase, as positive controls. After 16, 20 and 24 h of bloodfeeding, mosquitoes were dissected to extract the midgut and assess the effect of the different chitinases on membrane structure. Optical microscopy showed that formation of PM was clearly complete after 16 h in the posterior midgut from Anopheles already fed with healthy donor bloods. By contrast, PM formation was visible after 16 h in the posterior midgut of mosquitoes fed with malaria and Gaucher patient bloods but appeared clearly damaged at 20 and 24 h. At the same time, the PM formation was almost completely inhibited in the midgut of Anopheles fed with P. falciparum chitinase-enriched bloods. These alterations were clearly confirmed by transmission electronic microscopy. In the present paper, we demonstrate that human CHIT from different sources is active on anophelines' PM. PMID- 17136387 TI - Development and characterization of microsatellite markers for the canine hookworm, Ancylostoma caninum. AB - Microsatellites are repetitive genomic elements that show high levels of variation and therefore provide excellent tools to study the genetics of eukaryotic organisms. Hookworms are extremely common and important nematode parasites of humans and animals, causing potentially serious disease morbidity. Control of hookworms in dogs is achieved by frequent treatment with anthelmintics, and in humans, anthelmintics are frequently administered in a mass treatment community-wide approach. Understanding the population genetics of hookworms has important implications for studies on the development and spread of drug resistance. We investigated the genome of Ancylostoma caninum for microsatellites by developing and then screening an enriched genomic library as well as by data mining published sequences of a whole genome shotgun library. Investigations revealed a high abundance of trinucleotide repeats. Dinucleotide repeats were characterized by a high number of AT, GA, and GT repeats. After testing and optimization of 68 markers, a panel of 34 polymorphic microsatellite markers were selected. Microsatellite analysis of hookworm isolates revealed a high degree of polymorphism, which was not influenced by the length of the repeats. This panel of microsatellite markers makes it possible to pursue investigations on the population genetics of A. caninum. Furthermore, a number of the markers demonstrated suitability for analysis of the human hookworm species Necator americanus and A. duodenale. PMID- 17136388 TI - RNA interference of cytosolic leucine aminopeptidase reduces fecundity in the hard tick, Haemaphysalis longicornis. AB - Ticks are effective vectors of pathogens because of their blood feeding and high fecundity. This high fecundity is related to the size of the blood meal. Therefore, knowledge of how blood proteins are degraded and converted to proteins, including yolk protein, is important for the development of ways to inhibit the utilization of blood proteins by ticks. RNA interference (RNAi) is becoming a powerful post-transcriptional gene silencing technique that provides insight into gene function. We constructed a double-stranded RNA (dsRNA) based on a previously cloned Haemaphysalis longicornis leucine aminopeptidase (HlLAP) gene to reevaluate the biological role in tick blood digestion. Gene specific transcriptional, translational, and functional disruptions were achieved by the introduction of dsRNA into the ticks. Significantly delayed onset of egg-laying and reduced egg oviposition resulted from the RNAi for the HlLAP gene. These results suggest that HlLAP actually works as a blood digestive enzyme and affects tick fecundity via unknown mechanisms. The reduction of egg oviposition may be caused by a decrease in nutrients, especially free amino acids generated by HlLAP, from the blood meal. This is the first report of an impact on tick reproduction caused by gene silencing of a blood digestion-related molecule. PMID- 17136389 TI - Molecular cloning and characterization of a major egg antigen in Paragonimus westermani and its use in ELISA for the immunodiagnosis of paragonimiasis. AB - A recombinant protein of a Paragonimus westermani egg antigen was produced and tested as an antigen for the serologic diagnosis of P. westermani infection. The P. westermani egg antigen gene contains a single open reading frame of 966 base pairs encoding 322 amino acids from 5' methionine to the 3' stop codon. The predicted amino acid sequence of this egg antigen was 40, 38, and was 35% identical to heat shock proteins from Schistosoma japonicum, Schistosoma mansoni, and Taenia saginata. The distribution this antigen was investigated in adult worms by indirect immunofluorescence assay, and found to be distributed in eggs and uteri. The specificity and sensitivity of the recombinant antigen were assessed by enzyme-linked immunosorbent assay (ELISA) using sera from patients infected with different parasites, which included 41 patients with paragonimiasis, and negative controls. The diagnostic positive and negative predictive absorbance value was 0.24 and the sensitivity of ELISA using the recombinant antigen was 90.2%, and its specificity 100%. Our results suggest that the developed recombinant major egg antigen-based ELISA offers a highly sensitive and specific assay for the diagnosis of paragonimiasis. PMID- 17136390 TI - PTEN targeting: the search for novel insulin sensitisers provides new insight into obesity research. PMID- 17136391 TI - Children and adolescents with type 1 diabetes eat a more atherosclerosis-prone diet than healthy control subjects. AB - AIMS/HYPOTHESIS: We evaluated how well the diet of Norwegian children and adolescents with type 1 diabetes fulfils the Nordic and European dietary recommendations, focusing on parameters affecting prevention of atherosclerosis. We also compared the diet of this patient group with that of healthy same-age control subjects. MATERIALS AND METHODS: A total of 177 children and adolescents with type 1 diabetes (9-10-year-olds, 12-13-year-olds) and 1,809 healthy same-age control subjects recorded their food intake for 4 days in precoded food diaries. RESULTS: In children and adolescents with type 1 diabetes the percentage of energy (E%) from fat (33-35 E%) and saturated fat (14-15 E%) was higher than recommended for that group. Furthermore their intake of fibre was lower (16-19 g/day) than current recommendations. There were no differences in energy intake between diabetic subjects and healthy control subjects. Percentage of energy from fat (mean difference: 3.4 E%, p < 0.001) and saturated fat (mean difference: 1.0 E%, p < 0.001) was significantly higher among diabetic subjects than control subjects. Intake of fruits and vegetables was low (210 g/day) compared with recommendations, both in the diabetic and control subjects. CONCLUSIONS/INTERPRETATION: Diabetic children and adolescents had a high intake of energy from saturated fat and low intake of fibre, fruits and vegetables, which could increase the risk of development of atherosclerosis. This study supports the idea that nutritional guidance in the treatment of children and adolescents with type 1 diabetes should be more focused, especially with regard to intake of fibre, fruits and vegetables and to quality and quantity of fat intake. PMID- 17136392 TI - Circulating oxidised low-density lipoprotein and intercellular adhesion molecule 1 and risk of type 2 diabetes mellitus: the Atherosclerosis Risk in Communities Study. AB - AIMS/HYPOTHESIS: To evaluate the role of oxidative stress and inflammation in the aetiology of type 2 diabetes, we examined the association of oxidised LDL (ox LDL) and soluble intercellular adhesion molecule-1 (sICAM-1) levels with type 2 diabetes incidence over 9 years in the Atherosclerosis Risk in Communities Study. MATERIALS AND METHODS: In a large, prospective, case-cohort design, ox-LDL and sICAM-1 were measured in stored plasma samples collected at baseline in stratified samples of 581 diabetes cases and 572 non-cases selected from 10,275 middle-aged men and women without prevalent diabetes at baseline. RESULTS: Compared with non-cases, diabetes cases had significantly higher mean baseline levels of ox-LDL and sICAM-1. Elevated ox-LDL and sICAM-1 were both associated with increased risk of incident diabetes after adjustment for age, sex, race and centre, with hazard ratios for the highest vs lowest tertiles of 1.68 (95% CI 1.25-2.24) and 1.91 (95% CI 1.45-2.50), respectively. After additional adjustment for fasting glucose, waist circumference, HDL-cholesterol, triacylglycerol, hypertension and C-reactive protein, only sICAM-1 remained an independent predictor of incident diabetes (hazard ratio 1.50; 95% CI 1.02-2.23). CONCLUSIONS/INTERPRETATION: In this community-based cohort of middle-aged US adults, elevated plasma ox-LDL and sICAM-1 levels were associated with increased risk of type 2 diabetes. Measurement of ICAM-1 or ox-LDL, or other measures related to inflammation or oxidative stress, may be helpful in identifying those patient populations in which to test whether novel therapies that inhibit specific pathways related to inflammation or oxidative stress are beneficial in the prevention of diabetes in humans. PMID- 17136393 TI - Glucose inhibits glucagon secretion by a direct effect on mouse pancreatic alpha cells. AB - AIMS/HYPOTHESIS: The mechanisms by which glucose regulates glucagon release are poorly understood. The present study aimed to clarify the direct effects of glucose on the glucagon-releasing alpha cells and those effects mediated by paracrine islet factors. MATERIALS AND METHODS: Glucagon, insulin and somatostatin release were measured from incubated mouse pancreatic islets and the cytoplasmic Ca(2+) concentration ([Ca(2+)](i)) recorded in isolated mouse alpha cells. RESULTS: Glucose inhibited glucagon release with maximal effect at 7 mmol/l. Since this concentration corresponded to threshold stimulation of insulin secretion, it is unlikely that inhibition of glucagon secretion is mediated by beta cell factors. Although somatostatin secretion data seemed consistent with a role of this hormone in glucose-inhibited glucagon release, a somatostatin receptor type 2 antagonist stimulated glucagon release without diminishing the inhibitory effect of glucose. In islets exposed to tolbutamide plus 8 mmol/l K(+), glucose inhibited glucagon secretion without stimulating the release of insulin and somatostatin, indicating a direct inhibitory effect on the alpha cells that was independent of ATP-sensitive K(+) channels. Glucose lowered [Ca(2+)](i) of individual alpha cells independently of somatostatin and beta cell factors (insulin, Zn(2+) and gamma-aminobutyric acid). Glucose suppression of glucagon release was prevented by inhibitors of the sarco(endo)plasmic reticulum Ca(2+)-ATPase, which abolished the [Ca(2+)](i)-lowering effect of glucose on isolated alpha cells. CONCLUSIONS/INTERPRETATION: Beta cell factors or somatostatin do not seem to mediate glucose inhibition of glucagon secretion. We instead propose that glucose has a direct inhibitory effect on mouse alpha cells by suppressing a depolarising Ca(2+) store-operated current. PMID- 17136394 TI - Environmental stress increases skeletal fluctuating asymmetry in the moor frog Rana arvalis. AB - Whether fluctuating asymmetry (FA) provides a useful metric indicator of the degree of environmental stress experienced by populations is still a contentious issue. We investigated whether the degree of FA in skeletal elements is useful in elucidating the degree of environmental stress experienced by frog populations, and further, tested the proposition that a trait's sensitivity to stress--as reflected in the degree of FA--is related to the degree of directional selection experienced by the given trait. We compared the degree of FA in four bilateral skeletal elements of male and female moor frogs (Rana arvalis) originating from low (acidified) and neutral pH populations. While the degree of uncorrected FA was unrelated to the degree of acidity, the growth rate and age of the individuals, the size-corrected FA was significantly higher in low than in neutral pH populations and decreased with individual ages and growth rates. In addition, both measures of FA were significantly higher in males and in particular in traits presumably under high sexual selection as indicated by the degree of sexual size dimorphism. All in all, the results indicate that individuals from acidified localities are smaller, younger and exhibit a significantly higher degree of FA than individuals from neutral pH populations. These results constitute the first assessment of FA in amphibians and suggest that the degree of FA in skeletal traits can be a useful indicator of the degree of environmental stress experienced by amphibian populations. PMID- 17136395 TI - To avoid or deter: interactions among defensive and escape strategies in sabellid worms. AB - Numerous studies demonstrate how sessile marine organisms utilize chemical, structural, and nutritional deterrents to persist in predator-rich environments. Little is known, however, about how mobile, more behaviorally complex species minimize predation by integrating avoidance and deterrence strategies. We investigated this using sabellid polychaete worms from the Caribbean and temperate western Atlantic. Sabellids extend their feather-like radioles beyond their protective tubes for feeding and respiration; the body remains inside the tube and the radioles retract when threatened. We used co-occurring consumers to determine the palatability of radioles and bodies for each of the eight species tested. In addition, we examined chemical or structural traits affecting palatability and evaluated predator escape traits, such as tube strength, speed of radiole retraction, completeness of retraction, and sensitivity to a nearby disturbance. All species had unpalatable radioles that were chemically or structurally defended, but only two species had unpalatable bodies. Thus, most species allocated defenses to tissues that were most exposed to predation. The two species with chemically defended bodies, Bispira brunnea and Bispira variegata, relied less on behavioral escapes than the other species. Their tubes were weak, they did not retract until disturbances were very close, and B. brunnea retracted slowly and incompletely even when touched. Other species generally had stronger tubes and/or retracted when disturbances were farther away. This trade-off of deterrence versus escape even occurred within a single species when populations differed in palatability. Populations of B.variegata from North Carolina and Georgia were chemically deterrent to both temperate and tropical consumers, while populations from Panama and Florida were palatable. The more palatable Panama population retracted in response to distant movement, while the unpalatable North Carolina population did not retract until nearly touched. Thus, most species utilize a combination of predator avoidance and deterrence strategies, but more deterrent populations of species utilized avoidance less. PMID- 17136396 TI - Functional analysis of a novel potassium channel (KCNA1) mutation in hereditary myokymia. AB - Myokymia is characterized by spontaneous, involuntary muscle fiber group contraction visible as vermiform movement of the overlying skin. Myokymia with episodic ataxia is a rare, autosomal dominant trait caused by mutations in KCNA1, encoding a voltage-gated potassium channel. In the present study, we report a family with four members affected with myokymia. Additional clinical features included motor delay initially diagnosed as cerebral palsy, worsening with febrile illness, persistent extensor plantar reflex, and absence of epilepsy or episodic ataxia. Mutation analysis revealed a novel c.676C>A substitution in the potassium channel gene KCNA1, resulting in a T226K nonconservative missense mutation in the Kv1.1 subunit in all affected individuals. Electrophysiological studies of the mutant channel expressed in Xenopus oocytes indicated a loss of function. Co-expression of WT and mutant cRNAs significantly reduced whole-oocyte current compared to expression of WT Kv1.1 alone. PMID- 17136397 TI - Mutations of the LMNA gene can mimic autosomal dominant proximal spinal muscular atrophy. AB - The molecular basis of autosomal dominant spinal muscular atrophy (AD-SMA) is largely unknown. Because the phenotypic spectrum of diseases caused by LMNA mutations is extremely broad and includes myopathies, neuropathies, and cardiomyopathies designated as class 1 laminopathies, we sequenced the LMNA gene in index patients with the clinical picture of proximal SMA, who had a family history suggestive of autosomal dominant inheritance. Among the 19 families investigated, two showed pathogenic mutations of the LMNA gene, resulting in the diagnosis of a class 1 laminopathy in about 10% of our series. We found one novel truncating mutation (c.1477C > T, Q493X) and one previously described missense mutation (c.1130G > T, R377H) in the LMNA gene of two unrelated patients with adult-onset proximal SMA followed by cardiac involvement 14 and 22 years after the onset of weakness. The pedigrees of both families revealed a high frequency of cardiac abnormalities or sudden deaths. Our findings extend the spectrum of laminopathies and are of relevance for genetic counseling and clinical care of families presenting with adult-onset proximal SMA. Particularly, if neurogenic atrophy is combined with a cardiac disease in a family, this should prompt LMNA mutation analysis. PMID- 17136398 TI - Differential responsiveness to caffeine and perceived effects of caffeine in moderate and high regular caffeine consumers. AB - RATIONALE: Individual differences in responsiveness to caffeine occur even within a caffeine-consuming population, but the factors that mediate differential responsiveness remain unclear. OBJECTIVES: To compare caffeine's effects on performance and mood in a group of high vs moderate consumers of caffeine and to examine the potential role of subjective awareness of the effects of caffeine in mediating any differential responsiveness. MATERIALS AND METHODS: Two groups of regular caffeine consumers (<200 mg/day and >200 mg/day) attended two sessions at which mood and cognitive functions were measured before and 30 min after consumption of 400-mg caffeine or placebo in a capsule. Cognitive tests included visual information processing, match-to-sample visual search (MTS) and simple and choice reaction times. Post-session questionnaires asked participants to describe any perceived effect of capsule consumption. RESULTS: High consumers, but not moderate consumers, demonstrated significantly faster simple and choice reaction times after caffeine relative to placebo. These effects were not attributable to obvious group differences in withdrawal or tolerance because there were no group differences in baseline mood or in reports of negative affect after caffeine. Instead, the high consumers were more likely to report experiencing positive effects of caffeine, whereas the moderate consumers were more likely to report no effect. CONCLUSIONS: The sensitivity of caffeine consumers to the mood- and performance-enhancing effects of caffeine is related to their levels of habitual intake. High caffeine consumers are more likely than moderate consumers to perceive broadly positive effects of caffeine, and this may contribute to their levels of use. PMID- 17136399 TI - Effects of lofexidine on stress-induced and cue-induced opioid craving and opioid abstinence rates: preliminary findings. AB - OBJECTIVE: A preliminary study examined whether lofexidine decreases stress induced and cue-induced opioid craving and improves opioid abstinence in naltrexone-treated opioid-dependent individuals. MATERIALS AND METHODS: Eighteen opioid-dependent patients were stabilized for 4 weeks with naltrexone (50 mg daily) and lofexidine (2.4 mg bid) before entering a 4-week randomized, double blind placebo-controlled discontinuation study where one group continued on lofexidine for an additional 4 weeks, while the second was tapered to placebo (Lofexidine-naltrexone vs Placebo-naltrexone). Ten patients also participated in guided imagery exposure to stress, drug cue, and neutral scenarios in a single laboratory session. RESULTS: Lofexidine-naltrexone patients had higher opioid abstinence rates and improved relapse outcomes as compared to the Placebo naltrexone group. Furthermore, Lofexidine-naltrexone patients had significantly lower heart rates and an attenuated stress and drug cue-induced opioid craving response in the laboratory as compared to the Placebo-naltrexone group. CONCLUSIONS: Although preliminary, these findings are the first to document lofexidine's potential in addressing stress-related opioid craving and relapse outcomes in humans. The results also suggest that combination therapies that target both drug-related reinforcement (naltrexone) and stress- and cue-related aspects of drug seeking could be beneficial in addiction relapse prevention. Further development of lofexidine to address stress-related opioid craving and relapse is warranted. PMID- 17136400 TI - Increased accumbal dopamine during daily alcohol consumption and subsequent aggressive behavior in rats. AB - BACKGROUND: Alcohol drinking may lead to increased aggression in certain individuals, and both fighting and drinking increase levels of dopamine and serotonin in mesocorticolimbic structures. Assessing the dynamic changes in these neurotransmitters during the course of drinking and fighting has remained challenging. OBJECTIVE: The objective of the study was to learn about ongoing monoaminergic activity in the nucleus accumbens of rats that engaged in aggressive behavior after having consumed low doses of alcohol. MATERIALS AND METHODS: After male members of breeding pairs of Long-Evans rats displayed reliable aggression toward an intruder into their home cage, they were trained to consume a 10% alcohol solution, leading to blood alcohol levels of 20-80 mg/dl. Subsequently, the effect of daily alcohol self-administration on aggression was determined in biweekly confrontations with an intruder. Finally, rats were implanted with a microdialysis probe aimed at the n. accumbens for sample collection before, during, and after a 10-min alcohol drinking session followed by a 10-min aggressive confrontation. RESULTS: Accumbal dopamine, but not serotonin, levels tended to increase in anticipation of the daily alcohol session, reaching significance immediately after the alcohol session and remaining significantly elevated (by 40%) during and after the subsequent confrontation. No such changes were seen in residents that confronted an intruder without preceding alcohol consumption. Animals that had a history of becoming more aggressive after consumption of low levels of alcohol showed similar changes in dopamine levels as did animals that had no such history. CONCLUSIONS: The rise in accumbal dopamine confirms previous findings and seems to reflect the anticipation of alcohol consumption; it persisted during the aggressive confrontation regardless of the level of aggression. The daily alcohol drinking for several months may have facilitated dopamine release and masked any further changes associated with the aggressive encounter. PMID- 17136401 TI - Profile of executive deficits in cocaine and heroin polysubstance users: common and differential effects on separate executive components. AB - OBJECTIVES: Structure of executive function was examined and we contrasted performance of substance dependent individuals (polysubstance users) and control participants on neuropsychological measures assessing the different executive components obtained. Additionally, we contrasted performance of polysubstance users with preference for cocaine vs heroin and controls to explore possible differential effects of the main substance abused on executive impairment. METHODS: Two groups of participants were recruited: abstinent polysubstance users and controls. Polysubstance users were further subdivided based on their drug of choice (cocaine vs heroin). We administered to all participants a comprehensive protocol of executive measures, including tests of fluency, working memory, reasoning, inhibitory control, flexibility, and decision making. RESULTS: Consistent with previous models, the principal component analysis showed that executive functions are organized into four separate components, three of them previously described: updating, inhibition, and shifting; and a fourth component of decision making. Abstinent polysubstance users had clinically significant impairments on measures assessing these four executive components (with effect sizes ranging from 0.5 to 2.2). Cocaine polysubstance users had more severe impairments than heroin users and controls on measures of inhibition (Stroop) and shifting (go/no go and category test). Greater severity of drug use predicted poorer performance on updating measures. CONCLUSION: Executive functions can be fractionated into four relatively independent components. Chronic drug use is associated with widespread impairment of these four executive components, with cocaine use inducing more severe deficits on inhibition and shifting. These findings show both common and differential effects of two widely used drugs on different executive components. PMID- 17136402 TI - Methotrexate pharmacokinetics in infants with acute lymphoblastic leukemia. AB - PURPOSE: We performed a pharmacokinetic evaluation of methotrexate (MTX) in infants with acute lymphoblastic leukemia enrolled on the Pediatric Oncology Group (POG) 9407 Infant Leukemia Study to evaluate the effects of age on MTX pharmacokinetics and pharmacodynamics. METHODS: A pharmacokinetic database of 61 patients was developed by combining MTX data obtained from 16 patients in a pharmacokinetic sub-study with data obtained for clinical care in other patients enrolled on the POG 9407 protocol. The data were analyzed for the first dose of MTX given to patients in induction/intensification therapy. Patients received MTX (4 g/m2) over 24 h at week 4 of therapy. Toxicity data were also reviewed to evaluate the incidence of common MTX toxicities during the first 6 weeks of therapy (the induction/intensification phase). RESULTS: Steady-state clearance (mean+/-standard deviation) for infants aged 0-6 months was 89+/-32 ml/min/m2 compared to 111+/-40 for infants aged 7-12 months (P=0.030). In the subgroup of infants aged 0-3 months the mean steady-state clearance was 84+/-30 ml/min/m2 (P=0.026 vs. the 7-12-month group). The incidence of renal toxicity (all grades) during induction/intensification therapy was 23% in the 0-3 months age group compared to 0% (for n=27) in the group 7-12 months of age (P=0.029). There were no significant differences in hepatoxicity or mucous membrane toxicity between age groups. CONCLUSIONS: A modest difference in steady-state MTX clearance is observed between younger infants (0-6 months) and older infants (7-12 months). Very young infants (0-3 months) also experienced a slightly higher incidence of renal toxicity during induction/intensification therapy. Steady-state clearance for the older infants is similar to values reported for children in other studies. PMID- 17136403 TI - [Molecular imaging with new PET tracers]. AB - In the recent years, positron emission tomography (PET) has gained more and more importance, especially in oncology for primary staging, as well as for response evaluation. The glucose analogon (18)F-FDG is the most widely used tracer up to now. In this paper, we review the applications of newly developed, more specific PET tracers. These tracers allow for imaging of a variety of biological processes, such as hypoxia and proliferation. The expression of different receptors can be visualized, like the somatostatin receptor 2 and the integrin alphavbeta3. Moreover, gene expression can be imaged as well. While most of these approaches are currently in the first phases of clinical evaluation, imaging of hypoxia and proliferation might be integrated into the daily routine in the near future. PMID- 17136404 TI - [Molecular imaging--a new emphasis in radiology]. PMID- 17136405 TI - [Diagnostic value of multislice perfusion CT in dementia patients]. AB - BACKGROUND: The diagnostic value of perfusion CT in dementia patients was retrospectively evaluated in comparison to the Mini-Mental State Examination (MMSE). MATERIAL AND METHODS: The perfusion CT database of 55 subjects was investigated. All patients underwent an unenhanced, contrast-enhanced, and perfusion CT of the head. The clinical evaluation of the degree of dementia was performed with the MMSE. In the perfusion CT data, 24 ROI were marked. Then blood volume (BV), blood flow (BF), and mean transit time (MTT) were calculated and compared with the ipsilateral and contralateral regions and with the degree of dementia. RESULTS: With increasing degree of dementia, a significant decrease of the occipital and temporal BV was found. A significant decrease of the BF in the frontal lobe, basal ganglia, and occipital region was found. Concurrently, the MTT increased significantly in the basal ganglia region. The group with Alzheimer's disease showed significant regional hypoperfusion compared with the group of cognitively normal subjects in the frontal, basal ganglia, occipital, and temporal region. CONCLUSIONS: The cerebral perfusion decreased with increasing degree of dementia. The inexpensive and widely available perfusion CT reveals information about regional differences of cerebral perfusion, which may be useful in differentiating severity and types of dementia. PMID- 17136406 TI - Review of oral appliances for treatment of sleep-disordered breathing. AB - Between 1982 and 2006, there were 89 distinct publications dealing with oral appliance therapy involving a total of 3,027 patients, which reported results of sleep studies performed with and without the appliance. These studies, which constitute a very heterogeneous group in terms of methodology and patient population, are reviewed and the results summarized. This review focused on the following outcomes: sleep apnea (i.e. reduction in the apnea/hypopnea index or respiratory disturbance index), ability of oral appliances to reduce snoring, effect of oral appliances on daytime function, comparison of oral appliances with other treatments (continuous positive airway pressure and surgery), side effects, dental changes (overbite and overjet), and long-term compliance. We found that the success rate, defined as the ability of the oral appliances to reduce apnea/hypopnea index to less than 10, is 54%. The response rate, defined as at least 50% reduction in the initial apnea/hypopnea index (although it still remained above 10), is 21%. When only the results of randomized, crossover, placebo-controlled studies are considered, the success and response rates are 50% and 14%, respectively. Snoring was reduced by 45%. In the studies comparing oral appliances to continuous positive airway pressure (CPAP) or to uvulopalatopharyngoplasty (UPPP), an appliance reduced initial AHI by 42%, CPAP reduced it by 75%, and UPPP by 30%. The majority of patients prefer using oral appliance than CPAP. Use of oral appliances improves daytime function somewhat; the Epworth sleepiness score (ESS) dropped from 11.2 to 7.8 in 854 patients. A summary of the follow-up compliance data shows that at 30 months, 56-68% of patients continue to use oral appliance. Side effects are relatively minor but frequent. The most common ones are excessive salivation and teeth discomfort. Efficacy and side effects depend on the type of appliance, degree of protrusion, vertical opening, and other settings. We conclude that oral appliances, although not as effective as CPAP in reducing sleep apnea, snoring, and improving daytime function, have a definite role in the treatment of snoring and sleep apnea. PMID- 17136407 TI - Stage of primary infection with lymphocytic choriomeningitis virus determines predisposition or resistance of mice to secondary bacterial infections. AB - We investigated the effect of a primary non-lethal infection with lymphocytic choriomeningitis virus (LCMV) on the course and outcome of a secondary infection with the Gram-negative Salmonella enterica serovar Typhimurium or the Gram positive Listeria monocytogenes in mice. We found that at each stage of the viral infection the susceptibility of mice to bacterial super-infections changes dramatically and depends also on whether the secondary infection is a Gram positive or Gram-negative one. The study shows that the outcome of the secondary infection is determined by a delicate balance between the overproduction of and the hypersensitivity to inflammatory cytokines (TNF-alpha and IFN-gamma), as well as by the changes in blood leukocytes occurring in mice in the course of viral infection. PMID- 17136408 TI - Urinary elimination of nickel and cobalt in relation to airborne nickel and cobalt exposures in a battery plant. AB - OBJECTIVE: To estimate the relationship between Ni concentrations in the ambient air and in the urine, at a battery plant using nickel hydroxide. METHODS: Workers occupationally exposed to a mixture of nickel hydroxide, metallic cobalt and cobalt oxyhydroxide dust were studied during two consecutive workdays. Air levels of Ni and Co in total dust were determined by personal sampling in the breathing zone. Both metals in air were sampled by Teflon binder filters and analyzed by inductively coupled plasma absorption emission spectrophotometry. Urine was collected from 16 workers immediately before and after the work shift. Urinary Ni and Co concentrations were measured by electrothermal atomic absorption spectrometry. RESULTS: A poor correlation was seen between Co in the air and in post-shift urine (r = 0.491; P < 0.01), and no correlation was found between Ni in the air and in post-shift urine (r = 0.272; P = 0.15), probably due to the use of respiratory protection. The subjects were exposed to higher levels of Ni than Co (Ni (mg/m(3)) = -0.02 + 7.41 Co (mg/m(3)), r = 0.979, P < 0.0001). Thus, exposure to Co at 0.1 mg/m(3) should produce a Ni level of 0.7 mg/m(3). According to section XIII of the German list of MAK and BAT Values, a relationship between exposure to Co and urinary Co excretion, Co (microg/l) = 600 Co (mg/m(3)), has been established and the relationship between soluble or insoluble Ni salts in the air and Ni in urine was as follows: Ni (microg/l) = 10 + 600 Ni (mg/m(3)) or Ni (microg/l) = 7.5 + 75 Ni (mg/m(3)). Assuming nickel hydroxide to be soluble and to be insoluble, the Ni concentrations corresponding to Ni exposure at 0.7 mg/m(3) were calculated as 430 and 60 microg Ni/l, respectively. Similarly, exposure to Co at 0.1 mg/m(3) should result in Co urinary concentrations of 60 microg Co/l. On the other hand, a good correlation was found between Co and Ni in post-shift urine (Ni (microg/l) = 9.9 + 0.343 Co (microg/l), r = 0.833, P < 0.0001). On the basis of this relationship, the corresponding value found in our study was 0.343 x 60 microg Co/l + 9.9 = 30.5 microg Ni/l. This value was close to that calculated by the equation for a group of insoluble compounds, but about 14 times lower than that calculated by the equation for a group of soluble compounds. CONCLUSIONS: Our results suggest that exposure to nickel hydroxide yields lower urine nickel concentrations than the very soluble nickel salts, and that the grouping of nickel hydroxide might be reevaluated. Therefore, to evaluate conclusively the relationship between nickel hydroxide dust in the air and Ni in post-shift urine, further studies are necessary. PMID- 17136409 TI - Crucial role of lysosomal iron in the formation of dinitrosyl iron complexes in vivo. AB - Dinitrosyl non-heme-iron complexes (DNIC) are found in many nitric oxide producing tissues. A prerequisite of DNIC formation is the presence of nitric oxide, iron and thiol/imidazole groups. The aim of this study was to investigate the role of the cellular labile iron pool in the formation of DNIC in erythroid K562 cells. The cells were treated with a nitric oxide donor in the presence of a permeable (salicylaldehyde isonicotinoyl hydrazone) or a nonpermeable (desferrioxamine mesylate) iron chelator and DNIC formation was recorded using electron paramagnetic resonance. Both chelators inhibited DNIC formation up to 50% after 6 h of treatment. To further investigate the role of lysosomal iron in DNIC formation, we prevented lysosomal proteolysis by pretreatment of whole cells with NH4Cl. Pretreatment with NH4Cl inhibited the formation of DNIC in a time dependent manner that points to the importance of the degradation of iron metalloproteins in DNIC formation in vivo. Fractionation of the cell content after treatment with the nitric oxide donor revealed that DNIC is formed predominantly in the endosomal/lysosomal fraction. Taken together, these data indicate that lysosomal iron plays a crucial role in DNIC formation in vivo. Degradation of iron-containing metalloproteins seems to be important for this process. PMID- 17136410 TI - [Urological rehabilitation of spinal cord injury patients]. AB - The urological rehabilitation of spinal cord injury patients depends on an optimal urological treatment plan and good cooperation between the patient, general practitioner, urologist, and a centre that specialises in treating spinal cord injuries. Because of medical advancements in neuro-urology, one can assume that in cases of lifelong urological care, the individual's life expectancy will be almost normal. The recognition that nonphysiological bladder storage pressure results in restricted kidney function has led to various therapeutic strategies with complementary goals, such as protection of the upper urinary tract, urinary continence, individualized bladder management. PMID- 17136411 TI - Diagnostic value of 18F-FDG PET/CT in trauma patients with suspected chronic osteomyelitis. AB - PURPOSE: To retrospectively evaluate the diagnostic value of (18)F-FDG PET/CT in trauma patients with suspected chronic osteomyelitis. METHODS: Thirty-three partial body (18)F-FDG PET/CT scans were performed in 33 patients with trauma suspected of having chronic osteomyelitis. In 10 and 23 patients, infection was suspected in the axial and appendicular skeleton, respectively. In 18 patients, PET/CT was performed in the presence of metallic implants. Histopathology or bacteriological culture was used as the standard of reference. For statistical analysis, sensitivity, specificity and accuracy were calculated in relation to findings of the reference standard. RESULTS: Of 33 PET/CT scans, 17 were true positive, 13 true negative, two false positive and one false negative. Eighteen patients had chronic osteomyelitis and 15 had no osseous infection according to the reference standard. Sensitivity, specificity and accuracy for (18)F-FDG PET/CT was 94%, 87% and 91% for the whole group, 88%, 100% and 90% for the axial skeleton and 100%, 85% and 91% for the appendicular skeleton, respectively. CONCLUSION: (18)F-FDG PET/CT is a highly sensitive and specific method for the evaluation of chronic infection in the axial and appendicular skeleton in patients with trauma. PET/CT allows precise anatomical localisation and characterisation of the infectious focus and demonstrates the extent of chronic osteomyelitis with a high degree of accuracy. PMID- 17136412 TI - [Acute intermittent porphyria. A clinical chameleon: case study of a 40-year-old female patient]. AB - Acute porphyrias are rare, but often misdiagnosed and may take a dramatic clinical course. The combination of various internal, psychiatric and neurological symptoms can mimic different other diseases. We report a 40-year-old female patient who was admitted with a subacute tetraparesis. During the last 2 months the patient was treated several times because of abdominal pain and just before admission to our clinic in a psychiatric hospital because of acute mental changes and hallucinations. The typical combination of abdominal pain, motor neuropathy and psychiatric symptoms confirmed by increased amounts of porphyrins and their precursors, led us to promptly diagnose acute intermittent porphyria. Better knowledge about the pathogenesis has clearly improved the prognosis of acute porphyria. In remission, measurement of enzyme activities or mutation screening can be the only diagnostic verification. A mutation screening for family members should be conducted to identify symptom-free carriers, especially in cases of a positive family history. PMID- 17136413 TI - Immunolocalization of phospho-S6 kinases: a new way to detect mitosis in tissue sections and in cell culture. AB - During a study on the mTor pathway in the rat kidney we observed a striking increase of the phosphorylation of the S6 kinase in mitosis. In cryostat sections of perfusion-fixed tissue mitotic cells appeared as bright spots in immunofluorescence using an antibody specific for the phosphorylation site Thr421/Ser424. They were easily spotted in overviews with the objective 4x and 10x. Immunofluorescence was weak during the interphase. During the prophase it increased in both the nucleus and the cytoplasm and it remained bright during the subsequent phases of mitosis. All mitotic cells which were found in tubules and in the interstitium of the kidney using a chromatin stain displayed the bright immunofluorescence for phospho-S6 kinase. The same phenomenon was observed in rat liver and in mouse kidney as well as in a human cell line. Provided a rapid fixation, mitotic cells could be identified with the immunoperoxidase technique in paraffin sections of immersion-fixed tissue. This is the first report of phosphorylation of S6 kinase during mitosis in vivo. Thus, immunohistochemistry with anti-phospho-S6 kinase (Thr421/Ser424) appears to provide a convenient way to detect mitotic cells at low magnification. PMID- 17136414 TI - [Day-care laparoscopic cholecystectomy]. PMID- 17136415 TI - [Discrimination of musical pitch with cochlear implants]. AB - BACKGROUND: Numerous people with cochlear implants (CI) report difficulties in listening to music even though they understand speech quite well. One reason for this is a limited perception of pitch and timbre. In this study ability of adult CI subjects to discriminate musical pitch is investigated. PATIENTS AND METHODS: In two psychoacoustic experiments, each conducted in 10 adult CI subjects provided with MED-EL Combi 40+ cochlear implant devices and a control group of subjects with normal hearing, individual discrimination abilities for musical pitch perception were determined. To investigate the influence of the group of instruments on discrimination ability, stimuli representing four different groups of instruments were used: woodwind (clarinet), brass (trumpet), strings (violin) and keyboard instruments (piano). RESULTS: The discrimination thresholds determined varied between individual CI subjects, and on average they were significantly higher for the piano than for the other three instruments. CONCLUSIONS: The results show that in subjects with CI pitch perception differs from instrument to instrument and is in general worse than in persons with normal hearing. PMID- 17136416 TI - Spatial reorientation: the effects of space size on the encoding of landmark and geometry information. AB - The effects of the size of the environment on animals' spatial reorientation was investigated. Domestic chicks were trained to find food in a corner of either a small or a large rectangular enclosure. A distinctive panel was located at each of the four corners of the enclosures. After removal of the panels, chicks tested in the small enclosure showed better retention of geometrical information than chicks tested in the large enclosure. In contrast, after changing the enclosure from a rectangular-shaped to a square-shaped one, chicks tested in the large enclosure showed better retention of landmark (panels) information than chicks tested in the small enclosure. No differences in the encoding of the overall arrangement of landmarks were apparent when chicks were tested for generalisation in an enclosure differing from that of training in size together with a transformation (affine transformation) that altered the geometric relations between the target and the shape of the environment. These findings suggest that primacy of geometric or landmark information in reorientation tasks depends on the size of the experimental space, likely reflecting a preferential use of the most reliable source of information available during visual exploration of the environment. PMID- 17136418 TI - Are calcifying matrix vesicles in atherosclerotic lesions of cellular origin? AB - Over recent years, the role of matrix vesicles in the initial stages of arterial calcification has been recognized. Matrix calcifying vesicles have been isolated from atherosclerotic arteries and the biochemical composition of calcified vesicles has been studied. No studies have yet been carried out to examine the fine structure of matrix vesicles in order to visualize the features of the consequent stages of their calcification in arteries. In the present work, a high resolution ultrastructural analysis has been employed and the study revealed that matrix vesicles in human atherosclerotic lesions are heterogeneous with two main types which we classified. Type I calcified vesicles were presented by vesicles surrounded by two electron-dense layers and these vesicles were found to be resistant to the calcification process in atherosclerotic lesions in situ. Type II matrix vesicles were presented by vesicles surrounded by several electron dense layers and these vesicles were found to represent calcifying vesicles in atherosclerotic lesions. To test the hypothesis that calcification of matrix vesicles surrounded by multilayer sheets may occur simply as a physicochemical process, independently from the cell regulation, we produced multilamellar liposomes and induced their calcification in vitro in a manner similar to that occurring in matrix vesicles in atherosclerotic lesions in situ. PMID- 17136419 TI - Functional profile of activated dendritic cells in unstable atherosclerotic plaque. AB - BACKGROUND: Unstable atherosclerotic plaque typically contains an infiltrate of activated macrophages and activated T cells. This study established a functional profile of plaque-residing dendritic cells (DC) to examine whether they can function as Ag-presenting cells to facilitate in situ T-cell activation. METHODS: Carotid artery plaque tissues were collected from 19 asymptomatic and 38 symptomatic patients undergoing endarterectomy. Matched samples of normal coronary artery wall, stable nonruptured plaque, and eroded unstable plaque were harvested from patients with fatal myocardial infarction. Quantitative PCR and immunohistochemistry were used to analyze the tissues for markers of DC activation (CD83, CD86, CCL19,CCL21) and correlate them with T-cell activation (IFN-gamma,TNF-alpha). RESULTS: Carotid artery plaques from patients with ischemic symptoms compared to asymptomatic patients were characterized by the presence of high amount of T-cells (P<0.01) and tissue production of high levels of the T-cell cytokines IFN-gamma (P=0.001) and TNF-alpha (P=0.006). Plaque tissues from patients with ischemic complications contained elevated levels of CD83 (P<0.001), a marker of DC activation, and the DC chemokines CCL19 (P=0.001) and CCL21 (P<0.02). Unstable coronary artery plaques were similarly correlated compared to carotid plaques from symptomatic patients with the accumulation of T cells (P=0.001) and the production of T cell chemokines IFN-gamma (P=0.001) and TNF-alpha (P=0.002). Immunohistochemistry confirmed the presence of CD83(+) DC in the shoulder region of unstable plaques, where they produced the T cell attracting chemokines CCL19 and CCL21. Mapping of activated DC demonstrated close contact between mature DC and T cells expressing the activation marker CD40 ligand (CD40L). CONCLUSION: Activated and fully mature DC are represented in the inflammatory infiltrate characteristic for unstable carotid and coronary atheroma. Such DC produce chemokines, and thus can regulate the cell traffic into the lesion. Through the expression of the costimulatory ligand CD86, plaque residing DC can augment T-cell stimulation and provide optimal stimulation conditions for T lymphocytes, resembling the microenvironment in organized lymphoid tissues. PMID- 17136423 TI - AUREMOL-RFAC-3D, combination of R-factors and their use for automated quality assessment of protein solution structures. AB - We present here the computer program AUREMOL-RFAC-3D that is a generalization of the previously published program RFAC for the fully automated estimation of residual indices (R-factors) from 2D NOESY spectra. It is part of the larger AUREMOL software package (www.auremol.de). RFAC-3D calculates R-factors directly from two-dimensional homonuclear NOESY spectra as well as from three-dimensional (15)N or (13)C edited NOESY-HSQC spectra and thus extends the application range to larger proteins. The fully automated method includes automated peak picking and integration, a Bayesian noise and artifact recognition and the use of the complete relaxation matrix formalism. To enhance the reliability of the calculated R-factors the method is also generalized to calculate combined R factors from a set of 2D and 3D-spectra. For an optimal combination of the information derived from different sources a plausible formalism had to be derived. In addition, we present a novel direct R-factors based measure that correlates an R-factors as defined in this paper to the root mean square deviation of the actual structure from the optimal structure. The new program has been successfully tested on the histidine containing phosphocarrier protein (HPr) from Staphylococcus carnosus and on the Ras-binding domain (RBD) of the Ral guanine-nucleotide dissociation stimulation factor (RalGDS). PMID- 17136424 TI - Arabidopsis thaliana AtGppl and AtGpp2: two novel low molecular weight phosphatases involved in plant glycerol metabolism. AB - We have isolated two Arabidopsis thaliana genes, AtGppl and AtGpp2, showing homology with the yeast low molecular weight phosphatases GPP1 and GPP2, which have a high specificity for DL-glycerol-3-phosphate, and moreover homology with DOG1 and DOG2 that dephosphorylate 2-deoxyglucose-6-phosphate. Using a comparative genomic approach, the corresponding genes were identified as conceptual translated haloacid dehalogenase-like hydrolase proteins. AtGppl (gi 18416631) and AtGpp2 (gi 18423981), encode proteins that share 95% identity, with a predicted Mw of 33 and 27 kDa and a pI of 7.8 and 5.6, respectively. Both isoforms have a high specificity for DL-glycerol-3-phosphate, pH optima at 7.0, and Km in the range of 3.5-5.2 mM. AtGppl and AtGpp2 are expressed throughout development in all plant organs, most strongly in siliqua, and expression is not affected by osmotic, ionic or oxidative stress. A putative chloroplast transit peptide cTP-containing sequence is appended to the AtGppl N-terminus while AtGpp2, devoid of this tail, is predicted to be in the extraplastidial cytosol; this compartmenting was further confirmed by subcellular fractionation. An immunohystochemical localization study, using anti-AtGpp2 antibodies, indicates that the AtGpp proteins are mainly restricted to the meristem of immature flower and vascular elements of the root, shoot, leave, siliqua and developing embryo. Considerable immunoreaction was observed in the cytoplasm as well as in plastid compartments of distinct cells types from different heterotrophic Arabidopsis tissues, and particularly localised within phloem companion cells. Transgenic Arabidopsis plants, with gain of AtGpp2 function, show altered phosphatase activity rates and improved tolerance to salt, osmotic and oxidative stress. PMID- 17136425 TI - Load-induced focal adhesion mechanotransduction is altered with aging in the Fischer 344/NNiaHSd x Brown Norway/BiNia rat aorta. AB - The focal adhesion kinase (FAK) pathway has emerged as a critical component for mediating numerous cellular responses including control of cell growth, differentiation, and adaptation. Here we compared the expression, basal activation, and the ability of increased intraluminal pressure to activate FAK and focal adhesion-associated proteins in the aorta of adult (6 months old) and very aged (36 months old) Fischer 344/NNiaHSd x Brown Norway/BiNia (F344/NXBN) rats. Immunoblot analysis showed increases in the aortic content of FAK (15%), FAK related non-kinase (p41-FRNK) (28%), Src (92%), RhoA (41%), and paxillin (23%) in the very aged aortae. Increased age significantly changed the basal phosphorylation status of FAK and paxillin. Application of aortic intraluminal pressure (200 mm Hg) amplified the phosphorylation of FAK (Tyr 925), Src (Tyr 416), and paxillin (Tyr 188) in adult animals while aortic loading in the very aged animals failed to induce FAK (Tyr 925) phosphorylation. Aging did not alter the load-induced regulation of RhoA; however, FRNK (p41) translocation between cytosolic and membrane compartments was increased. These results confirm previous observations that FAK and focal adhesion-associated proteins are mechanically regulated and expand these studies to suggest that FAK mechanotransduction is altered with aging. PMID- 17136426 TI - Fragile x mental retardation (Fmr-1) gene expression is down regulated in brain of mice during aging. AB - Fragile x syndrome (FXS) is the most common form of inherited mental retardation disease. This is caused due to expansion of CGG triplet in 5'-untranslated region of fragile x mental retardation 1 (FMR-1) gene. In most of the cases, abnormally large size of the CGG repeat (>200) undergoes hypermethylation, which in turn silences the FMR-1 gene causing thereby complete lack of its protein product called fragile x mental retardation protein (FMRP). Lack of FMRP due to gene silencing or production of faulty protein due to point mutation in KH2 domain of FMRP alters the translational process in neurons and leads to expression of mental retardation phenotype on the patients. The FMRP is expressed ubiquitously in all tissues; however, it is predominantly expressed in neurons and testis. It possesses heterogeneity and is found in many isoforms due to alternative splicing of the FMR-1 transcript. Based on our data from the Western-, slot-, Northern blotting and immunohistochemical studies, we report here the down regulation of Fmr-1 gene and FMRP in mice brain in age-dependent manner. The present finding is important in respect to FMRP-dependent various brain functions i.e., learning, memory, cognition etc. that decrease with advancing age. PMID- 17136427 TI - Heliconema longissimum (Ortlepp, 1923) (Nematoda: Physalopteridae) from Pisodonophis boro (Teleostei: Ophichthidae) in Thailand, with remarks on the taxonomy of the Proleptinae Schulz, 1927. AB - Physalopterid nematodes identified as Heliconema longissimum (Ortlepp, 1923) were collected from the stomach of rice-paddy eels Pisodonophis boro (Hamilton) (Anguilliformes: Ophichthidae) from two brackish-water localities (mangroves) in Thailand: one in Phan-Nga Province, southwestern Thailand, northeast of Phuket Island, and one in Ranong Province, near the border with Myanmar. Study of the morphology of this hitherto insufficiently known nematode species, including its first SEM examination, enabled a detailed redescription of H. longissimum. Present taxonomic problems in the subfamily Proleptinae Schulz, 1927 are discussed, where a new delimitation of Proleptus Dujardin, 1845, Heliconema Travassos, 1919 and Paraleptus Wu, 1927 is proposed based on the cephalic dentation. H. minnanensis [sic] Damin & Heqing, 2001 is transferred to Paraleptus Wu, 1927 as P. minnanensis (Damin & Heqing, 2001) n. comb. and Paraleptus chiloscyllii Yin & Zhang, 1983 transferred by Damin & Heqing (2001) to Heliconema, is retained in Paraleptus. H. ahiri Karve, 1941 is considered a junior synonym of H. longissimum (Ortlepp, 1923). The present finding of H. longissimum in Pisodonophis boro represents the first host record and the first record of this nematode from Thailand. PMID- 17136428 TI - Looking for the primordial genetic honeycomb. AB - All life forms on Earth share the same biological program based on the DNA/RNA genomes and proteins. The genetic information, recorded in the nucleotide sequence of the DNA and RNA molecule, supplies the language of life which is transferred through the different generations, thus ensuring the perpetuation of genetic information on Earth. The presence of a genetic system is absolutely essential to life. Thus, the appearance in an ancestral era of a nucleic acid like polymer able to undergo Darwinian evolution indicates the beginning of life on our planet. The building of primordial genetic molecules, whatever they were, required the presence of a protected environment, allowing the synthesis and concentration of precursors (nucleotides), their joining into larger molecules (polynucleotides), the protection of forming polymers against degradation (i.e. by cosmic and UV radiation), thus ensuring their persistence in a changing environment, and the expression of the "biological" potential of the molecule (its capacity to self-replicate and evolve). Determining how these steps occurred and how the primordial genetic molecules originated on Earth is a very difficult problem that still must be resolved. It has long been proposed that surface chemistry, i.e. on clay minerals, could have played a crucial role in the prebiotic formation of molecules basic to life. In the present work, we discuss results obtained in different fields that strengthen the hypothesis of a clay surface-mediated origin of genetic material. PMID- 17136429 TI - About a formamide-based origin of informational polymers: syntheses of nucleobases and favourable thermodynamic niches for early polymers. AB - Formamide NH(2)CHO chemistry provides a unitary frame into which several pieces of the origin-of-life puzzle may be adjusted. Synthetic processes were uncovered which, starting from formamide and prebiotically easily available common catalysts, yield all the necessary nucleic bases precursors, including acyclonucleosides. Formamide allows phosphorylations and trans-phosphorylations, favours the micellar aggregation of surfactants and, most importantly, determines conditions in which the formation of nucleic polymers is thermodynamically favoured. In the detected conditions, the phosphoester bonds are more stable in the polymeric than in the monomeric form, thus allowing formation and survival of informational nucleic polymers. PMID- 17136430 TI - A new miniaturised optical system for chemical species spectroscopic detection based on a scanning integrated Mach-Zehnder microinterferometer on LiNbO3. AB - Absorption or emission spectroscopy is a powerful tool for detecting chemical compounds, diluted in fluid media: the sensitivity of this technique depends on the optical path of the source radiation, on the spectral window used for analysis and on the spectrometer performances. In this view, we designed and produced the first prototypes of an integrated scanning Fourier Transform Microinterferometer with Mach-Zehnder geometry, by using MEOS (Micro Electro Optical Systems) technologies. The microdevice, obtained by fabricating integrated optical waveguides on LiNbO(3) (LN) crystals, is electrically driven, without moving parts, by exploiting the electrooptical properties of the material. The microdevice operates the Fourier Transform of the input radiation spectral distribution, which can be reconstructed starting from the output signal by means of Fast Fourier Transform (FFT) techniques. The microinterferometer weights few grams, the power consumption is of a few mW and, in principle, can operate in the LN transmittance range (0.36-4.5 microm). The microinterferometer performances were preliminary tested in the (0.4-1.7 microm) spectral window. In the Visible region (0.4-0.7 mum) this microsystem demonstrated a spectral resolution suitable for detecting the characteristic lines of the solar spectrum together with the absorption bands of common gases present in Earth's atmosphere. In a further experiment we tested its performances for gas trace detection by using a calibrated NO(2) optical gas cell, showing the possibility to reveal up to 10 ppb, when suitable optical paths are used. Finally, colorimetry tests for the titration of an organic dye (E131) in alcohol solution are presented. PMID- 17136431 TI - Hydrothermal reactions of pyruvic acid: synthesis, selection, and self-assembly of amphiphilic molecules. AB - Selection and self-assembly of organic compounds in aqueous phases must have been a primary process leading to emergent molecular complexity and ultimately to the origin of life. Facile reactions of pyruvic acid under hydrothermal conditions produce a complex mixture of larger organic molecules, some of which are amphiphiles that readily self-assemble into cell-sized vesicular structures. Chemical characterization of major components of this mixture reveals similarities to the suite of organic compounds present in the Murchison carbonaceous chondrite, some of whose molecules also self-assemble into membranous vesicles. Physical properties of the products are thus relevant to understanding the prebiotic emergence of molecular complexity. These results suggest that a robust family of prebiotic reaction pathways produces similar products over a range of geochemical and astrochemical environments. PMID- 17136432 TI - Prebiotic microreactors: a synthesis of purines and dihydroxy compounds in aqueous aerosol. AB - We report the synthesis of purine bases and other heterocycles and the formation of amino acids, hydroxy acids and dihydroxy compounds by the spark activation of an atmosphere of methane, nitrogen and hydrogen, in the presence of an aqueous aerosol. With the aid of the interface air-water, the organic material obtained shows greater amounts and diversity of molecules with biological interest than the products obtained in the absence of an aerosol. Our results support the suggestion that aerosols may have played a significant role in the prebiotic origin of molecular diversity and evolution. PMID- 17136434 TI - A possible path to the RNA world: enantioselective and diastereoselective purification of ribose. AB - A theoretical mechanism resulting in the prebiotic appearance of enantiopure ribose, which would be needed for the origin of RNA and the "RNA world" is proposed. The mechanism simultaneously explains the emergence of biological homochirality and could answer the question of why nucleic acids are based on ribose rather than another sugar. Cleavage of certain non-chiral mineral crystals is known to lead to formation of chiral surfaces. In a chromatography-like process a mixture of racemic carbohydrates originating from the formose reaction is proposed to have been separated on such a chiral surface. Monosaccharides interact with surfaces through their hydroxyl groups, either by hydrogen bond formation or complex formation with metal ions. alpha-Ribopyranose, which has all hydroxyl groups on one side of the ring, is known to interact more strongly than other sugars with surfaces, as corroborated by certain chromatographic and electrophoresis data. A similar scenario leading to enantiopure ribose is separation on a flat, but not necessarily chiral surface in the presence of a strong electric field capable of orienting highly polar derivatives of sugars. PMID- 17136435 TI - Microbial community of a hydrothermal mud vent underneath the deep-sea anoxic brine lake Urania (eastern Mediterranean). AB - The composition of a metabolically active prokaryotic community thriving in hydrothermal mud fluids of the deep-sea hypersaline anoxic Western Urania Basin was characterized using rRNA-based phylogenetic analysis of a clone library. The physiologically active prokaryotic assemblage in this extreme environment showed a great genetic diversity. Most members of the microbial community appeared to be affiliated to yet uncultured organisms from similar ecosystems, i.e., deep-sea hypersaline basins and hydrothermal vents. The bacterial clone library was dominated by phylotypes affiliated with the epsilon-Proteobacteria subdivision recognized as an ecologically significant group of bacteria inhabiting deep-sea hydrothermal environments. Almost 18% of all bacterial clones were related to delta-Proteobacteria, suggesting that sulfate reduction is one of the dominant metabolic processes occurring in warm mud fluids. The remaining bacterial phylotypes were related to alpha- and beta-Proteobacteria, Actinobacteria, Bacteroides, Deinococcus-Thermus, KB1 and OP-11 candidate divisions. Moreover, a novel monophyletic clade, deeply branched with unaffiliated 16S rDNA clones was also retrieved from deep-sea sediments and halocline of Urania Basin. Archaeal diversity was much lower and detected phylotypes included organisms affiliated exclusively with the Euryarchaeota. More than 96% of the archaeal clones belonged to the MSBL-1 candidate order recently found in hypersaline anoxic environments, such as endoevaporitic microbial mats, Mediterranean deep-sea mud volcanoes and anoxic basins. Two phylotypes, represented by single clones were related to uncultured groups DHVE-1 and ANME-1. Thus, the hydrothermal mud of hypersaline Urania Basin seems to contain new microbial diversity. The prokaryotic community was significantly different from that occurring in the upper layers of the Urania Basin since 60% of all bacterial and 40% of all archaeal phylotypes were obtained only from mud fluids. The uniqueness of the composition of the active prokaryotic community could be explained by the complex environmental conditions at the site. The interaction of oxygenated warm mud fluids with the cold hypersaline brine of the Urania Basin seems to simultaneously select for various metabolic processes, such as aerobic and anaerobic heterotrophy, sulfide- and methane-dependent chemotrophy along with anaerobic oxidation of methane, sulfate- and metal reduction. PMID- 17136436 TI - Phosphate solubility and the cyanate-mediated synthesis of pyrophosphate. AB - The justification for a less alkaline primordial ocean (than present) is briefly reviewed, along with constraints on aqueous phosphate under such conditions. Based on the assumption that CaHPO(4) dihydrate determined the availability of phosphorus species, we have carried out laboratory simulations to determine equilibrium concentrations as a function of pH (in PIPES buffer) with added NaCl and CaCl(2). Consistent with expectations, solubility declines with higher pH and [CaCl(2)], but increases only slightly with [NaCl]. Significantly, PIPES shows no specific effect on the dissolution beyond its influence on pH and ionic strength. Data are also presented on the synthesis of pyrophosphate from the NaOCN/CaHPO(4).2H(2)O system, which could have provided a source of this phosphate anhydride on the early Earth. PMID- 17136437 TI - The viability of a nonenzymatic reductive citric acid cycle--kinetics and thermochemistry. AB - The likelihood of a functioning nonenzymatic reductive citric acid cycle, recently proposed as the precursor to biosynthesis on early Earth, is examined on the basis of the kinetics and thermochemistry of the acetate --> pyruvate --> oxaloacetate --> malate sequence. Using data derived from studies of the Pd catalyzed phosphinate reduction of carbonyl functions it is shown that the rate of conversion of pyruvate to malate with that system would have been much too slow to have played a role in the early chemistry of life, while naturally occurring reduction systems such as the fayalite-magnetite-quartz and pyrrhotite pyrite-magnetite mineral assemblages would have provided even slower conversions. It is also shown that the production of pyruvate from acetate is too highly endoergic to be driven by a naturally occurring energy source such as pyrophosphate. It is thus highly doubtful that the cycle can operate at suitable rates without enzymes, and most unlikely that it could have participated in the chemistry leading to life. PMID- 17136438 TI - Towards a Confucian virtue bioethics: reframing Chinese medical ethics in a market economy. AB - This essay addresses a moral and cultural challenge facing health care in the People's Republic of China: the need to create an understanding of medical professionalism that recognizes the new economic realities of China and that can maintain the integrity of the medical profession. It examines the rich Confucian resources for bioethics and health care policy by focusing on the Confucian tradition's account of how virtue and human flourishing are compatible with the pursuit of profit. It offers the Confucian account of the division of labor and the financial inequalities this produces with special attention to China's socialist project of creating the profession of barefoot doctors as egalitarian peasant physicians and why this project failed. It then further develops the Confucian acknowledgement of the unequal value of different services and products and how this conflicts with the current system of payment to physicians which has led to the corruption of medical professionalism through illegal supplementary payments. It further gives an account the oblique intentionality of Confucian moral psychology that shows how virtuous persons can pursue benevolent actions while both foreseeing profit and avoiding defining their character by greed. This account of Confucian virtue offers the basis for a medical professionalism that can function morally within a robustly profit-oriented market economy. The paper concludes with a summary of the characteristics of Confucian medical professionalism and of how it places the profit motive within its account of virtue ethics. PMID- 17136439 TI - Arachidonic acid hydroperoxide stimulates lipid peroxidation in rat liver nuclei and chromatin fractions. AB - Arachidonic acid, the most abundant polyunsaturated fatty acid in rat liver nuclei phospholipids is a major target of free radical attack, which induces lipid peroxidation. The non-enzymatic lipid peroxidation process in intact rat liver nuclei and in several chromatin fractions indicated that the most sensitive fatty acid for peroxidation is arachidonic acid C20:4 n-6. In this study, the effect of different amounts of arachidonic acid hydroperoxide on the lipid peroxidation of rat liver nuclei and chromatin fractions was studied; rat liver nuclei and chromatin fractions deprived of exogenous added hydroperoxide were utilized as control. The addition of arachidonic acid hydroperoxide to liver nuclei produces a marked increase in light emission that was hydroperoxide concentration dependent. The maximal peak of chemiluminescence displayed by the different chromatin fractions analyzed was observed between 20 and 80 min of incubation. The highest value of light emission was displayed by the high-density chromatin fractions, the 27.5 K fraction showed intermediate values of light emission, whereas the lowest density fraction produced very low chemiluminescence. A high correlation between arachidonic acid hydroperoxide concentration and chemiluminescence in the different chromatin fractions was observed. PMID- 17136440 TI - Premature aging in uremia. AB - Guanidinosuccinic acid is an aberrant metabolite isolated 40 years ago in the blood and urine of uremic subjects and a suspect in the toxicity associated with renal failure. It plays a minor role in the bleeding diathesis of uremia, contributes to the methyl group deficiency of dialysis patients, and is a factor in the premature atherosclerosis of end stage renal disease through the induction of hyperhomocysteinemia. As a major player, however, in the diversity and severity of uremic symptoms, it is a disappointment. Recently its source has been identified. It results from the superoxidation of argininosuccinic acid, which leads, also, to the production of gamma glutamic semialdehyde, an advanced glycation end product (AGE), which normally results from from the Maillard reaction, the non-enzymatic browning of protein. AGEs stimulate cross-linkages in protein that lead ultimately to loss of function, phagocytosis, and removal, and are important elements in the premature aging characteristic of renal disease, and diabetes. PMID- 17136441 TI - MAP kinases: differential activation following in vivo and ex vivo irradiation. AB - Mitogen activated protein kinases (MAPK) play a critical role in controlling cell survival and repopulation following exposure to ionising radiation. Most investigations on these pathways have been done using cultured cells or by ex vivo treatments. The present study was carried out to determine whether the response of MAPKs in mouse lymphocytes differs following in vivo and ex vivo irradiation with 60Co gamma-rays. We observed that ex vivo treatment resulted in a very significant decrease in the activated p44/42 and p38 MAPK as compared to in vivo. However, stress activated protein kinase (SAPK) response showed no significant difference between in vivo and ex vivo treatments. These observations point towards the differences in response elicited when the treatment is given in vivo as compared to in vitro. Therefore the findings reported from in vitro or ex vivo treatments should be treated with caution especially if it has to be clinically applied. PMID- 17136442 TI - Effects of repeated desflurane and sevoflurane anesthesia on enzymatic free radical scavanger system. AB - BACKGROUND: To investigate the possible effects of repeated sevoflurane and desflurane anesthesia on hepatocellular system by evaluating the free radical metabolism, hepatocellular enzymes and histopatholgical changes in rats. METHODS: Four groups of animals were studied. Sevoflurane 2% (v/v) and desflurane 6% (v/v) in air/O2 were administered to animals in group II (n=9) and III (n=9) respectively. 100% (v/v) O(2) was administered in group IV (n=9). Administration was done for 60 minutes over 3 days. Nine animals were allocated to control group (group I), superoxide dismutase (SOD), catalase (CAT), glutathion peroxidase (GSH Px), glutathione-s-transferase (GST) and thiobarbituric acid reactive substances (TBARS) were studied. Also electron microscopy was performed. RESULTS: Catalase, SOD, GSH-Px, GST activities and TBARS levels were significantly higher in groups II and III than in group I (p<0.05). All parameters were significantly higher in groups II versus group IV (p<0.05). On the other hand, SOD, GSH-Px and GST activities were significantly elevated in group III than IV, but CAT activity and TBARS levels were not significantly. Catalase, SOD, GSH-Px, GST but not TBARS levels were significantly higher in groups II and III than in group IV (p<0.05). TBARS levels were higher in group III than in group IV, but this elevation was not statistically significant. CAT, SOD and GSH-Px activities were significantly higher in groups II than in group III (p<0.05). CONCLUSION: Although electron microscopy findings were similar for group II and III, we can conclude that sevoflurane might cause more cellular damage than desflurane by causing higher activation of free radical metabolising enzymes. PMID- 17136443 TI - Exercise-induced stress enhances mammary tumor growth in rats: beneficial effect of the hormone melatonin. AB - We hypothesized that intense exercise training (forced swimming for 30 min, 5 days/week) may enhance the progression of mammary carcinogenesis through the involvement of stress hormones, such as catecholamines and prolactin, which can promote breast cancer. After the appearance of the DMBA-induced tumors in Sprague Dawley rats, the effect was evaluated of exercise-induced stress (with or without administration of the hormone melatonin) on the survival time, tumor multiplicity, and tumor growth until the death of the animals. In a second set of experiments, after one month of exercise, the NK cells count in blood, and the plasma concentrations of catecholamines and prolactin were determined. Although no significant change was found in either the survival time of the rats or the tumor multiplicity, exercise significantly increased the tumor growth rate. Stress was confirmed by the enhanced adrenaline and prolactin concentrations in the blood of the exercised rats. Exercise-induced stress did not change the percentage of NK cells in the tumor-bearing rats. Melatonin counteracted the increased tumor growth, returning the prolactin and adrenaline concentrations to their optimal physiological levels in the exercised tumor-bearing rats, thus confirming an "anti-stress" role of this hormone. In conclusion, intense exercise induced stress enhances mammary carcinogenesis through the involvement of adrenaline and prolactin. The results also confirmed a role of melatonin as a therapeutic aid against breast cancer in general, and in particular during situations of stress. PMID- 17136444 TI - Diazoxide-induced respiratory inhibition - a putative mitochondrial K(ATP) channel independent mechanism of pharmacological preconditioning. AB - The ischemic preconditioning biological phenomenon has been explored to identify putative pharmacologic agents to mimic this cytoprotective program against cellular ischemic injury. Diazoxide administration confers this cytoprotection, however, whether this is via direct activation of the putative mitochondrial K(ATP) (mK(ATP)) channel which was originally proposed has been questioned. Here, we present data supporting an alternate hypothesis evoking mitochondrial respiratory inhibition rather than mK(ATP) channel activation, as a mediating event in the diazoxide-activated cytoprotective program. Mitochondrial respiration and reactive oxygen species (ROS) production was measured in digitonin-permeabilized C2C12 myotubes, allowing for the modulation of mK(ATP) conductance by changing the potassium concentration of the medium (0-130 mM). Diazoxide dose-dependently attenuated succinate-supported respiration, an effect that was independent of mK(ATP) channel conductance. Similarly, 5 hydroxydecanoate (5-HD), a putative mK(ATP) channel blocker, released diazoxide induced respiratory inhibition independently of potassium concentration. Since diazoxide-induced cytoprotection and respiratory inhibition are both integrally linked to ROS generation we repeated above experiments following ROS generation using DCF fluorescence. Cytoprotective doses of diazoxide increased ROS generation independently of potassium concentration and 5-HD inhibited ROS production under the same conditions. Collectively these data support the hypothesis that diazoxide-mediated cytoprotection is independent of the conductance of the mK(ATP) channel and rather implicate mitochondrial respiratory inhibition-triggered ROS signaling. PMID- 17136445 TI - The relative contributions of compression and hypoxia to development of muscle tissue damage: an in vitro study. AB - Deep pressure ulcers develop in tissues subjected to sustained mechanical loading. Though it has been hypothesized that this damage mechanism results from local tissue ischemia, it has recently been shown with a cell model that sustained compression can cause cell deformation, leading to tissue breakdown. The present study focuses on the assessment of cell viability during compression and ischemia in an in vitro muscle model to determine their relative contributions to damage development. A model system was developed consisting of engineered skeletal muscle produced from the culture of murine muscle cells in a collagen gel. The tissue was subjected to 0, 20, or 40% compression under hypoxic or normoxic conditions. Experiments were performed on the stage of a microscope and cell viability was monitored using fluorescent markers for apoptotic and necrotic cell death. Hypoxia did not lead to significant cell death over a 22 h period. By contrast, compression led to immediate cell death that increased with time. No additional effect of hypoxia on cell death was observed. These data show that contrary to existing theories, compression can cause development of muscle damage and that hypoxia does not contribute to cell death development within 22 h in engineered muscle. PMID- 17136446 TI - Use of X-ray tomography to map crystalline and amorphous phases in frozen biomaterials. AB - The outcome of both cryopreservation and cryosurgical freezing applications is influenced by the concentration and type of the cryoprotective agent (CPA) or the cryodestructive agent (i.e., the chemical adjuvants referred to here as CDA) added prior to freezing. It also depends on the amount and type of crystalline, amorphous and/or eutectic phases formed during freezing which can differentially affect viability. This work describes the use of X-ray computer tomography (CT) for non-invasive, indirect determination of the phase, solute concentration and temperature within biomaterials (CPA, CDA loaded solutions and tissues) by X-ray attenuation before and after freezing. Specifically, this work focuses on establishing the feasibility of CT (100-420 kV acceleration voltage) to accurately measure the concentration of glycerol or salt as model CPA and CDAs in unfrozen solutions and tissues at 20 degrees C, or the phase in frozen solutions and tissue systems at -78.5 and -196 degrees C. The solutions are composed of water with physiological concentrations of NaCl (0.88% wt/wt) and DMEM (Dulbecco's Modified Eagle's Medium) with added glycerol (0-8 M). The tissue system is chosen as 3 mm thick porcine liver slices as well as 2 cm diameter cores which were either imaged fresh (3-4 h cold ischemia) or after loading with DMEM based glycerol solutions (0-8 M) for times ranging from hours to 7 days at 4 degrees C. The X-ray attenuation is reported in Hounsfield units (HU), a clinical measurement which normalizes X-ray attenuation values by the difference between those of water and air. NaCl solutions from 0 to 23.3% wt/wt (i.e. water to eutectic concentration) were found to linearly correspond to HU in a range from 0 to 155. At -196 degrees C the variation was from -80 to 95 HU while at -78.5 degrees C all readings were roughly 10 HU lower. At 20 degrees C NaCl and DMEM solutions with 0-8 M glycerol loading show a linear variation from 0 to 145 HU. After freezing to -78.5 degrees C the variation of the NaCl and DMEM solutions is more than twice as large between -90 and +190 HU and was distinctly non-linear above 6 M. After freezing to -196 degrees C the variation of the NaCl and DMEM solutions increased even further to -80 to +225 HU and was distinctly non-linear above 4 M, which after modeling the phase change and crystallization process is shown to correlate with an amorphous phase. In all tissue systems the HU readings were similar to solutions but higher by roughly 30 HU, as well as showing some deviations at 0 M after storage, probably due to tissue swelling. The standard deviations in all measurements were roughly 5 HU or below in all samples. In addition, two practical examples for CT use were demonstrated including: (1) glycerol loading and freezing of tissue cores and, (2) a mock cryosurgical procedure. In the loading experiment CT was able to measure the permeation of the glycerol into the sample at 20 degrees C, as well as the evolution of distinct amorphous vs. crystalline phases after freezing to -196 degrees C. In the mock cryosurgery example, the iceball edge was clearly visualized, and attempts to determine the temperature within the iceball are discussed. An added benefit of this work is that the density of these frozen samples, an essential property in measurement and modeling of thermal processes, was obtained in comparison to ice. PMID- 17136447 TI - New York State physicians: characteristics and distribution in health professional shortage areas. PMID- 17136448 TI - The President's New Freedom Commission: capitalizing on opportunities to advance school-based mental health services. AB - The report from President George W. Bush's New Freedom Commission on Mental Health (NFC), Achieving the Promise: Transforming Mental Health Care in America(2003), proposes goals and recommendations for improving mental health services. This report has significant implications for the delivery of mental health services through the schools. A focused discussion of the potential opportunities and challenges of implementing NFC recommendations related to school-based mental health is presented. Strategies for addressing five key areas at the intersection of school mental health and the Commission's recommendations include: stigma reduction, suicide prevention, expansion and improvement of school mental health, and screening and treatment of co-occurring mental health and substance abuse disorders. PMID- 17136449 TI - Population pharmacokinetics of risperidone and 9-hydroxyrisperidone in patients with acute episodes associated with bipolar I disorder. AB - A population model was developed with the aim to simultaneously describe risperidone and 9-hydroxyrisperidone pharmacokinetics; to obtain estimates for pharmacokinetic parameters and associated inter- and intra-individual variability of risperidone and 9-hydroxyrisperidone; and to evaluate the influence of patient demographic characteristics and other factors on risperidone, 9 hydroxyrisperidone, and active moiety pharmacokinetics. Data were obtained from 407 patients enrolled in four Phase 1 (serial blood sampling) and three Phase 3 trials (sparse sampling), representing dosage regimens ranging from 4 mg single dose to flexible 1-6 mg once daily. A pharmacokinetic model with two-compartment submodels for risperidone and 9-hydroxyrisperidone disposition and a sequential zero- and first-order absorption pathway was selected based on prior knowledge. A mixture model was incorporated due to CYP2D6 polymorphism of risperidone conversion to 9-hydroxyrisperidone. Patient characteristics tested as potential covariates were: age, sex, race, body weight, lean body mass, body mass index, creatinine clearance, liver function laboratory parameters, study, and carbamazepine comedication. The quasi-clearance of active moiety (the sum of risperidone and 9-hydroxyrisperidone) was simulated and linear regression performed to identify significant covariates. The selected pharmacokinetic model described the plasma concentration-time profiles for risperidone and 9 hydroxyrisperidone quite well and was able to determine each patient's phenotype. Covariates significantly affecting the pharmacokinetics were carbamazepine comedication, and study because the proportion of patients assigned to the intermediate metabolizer status decreased from single to multiple dosing while the proportion assigned to extensive metabolizer status increased. Covariates with limited and clinically irrelevant effects on active moiety concentrations were patient phenotype, race, and total protein. Carbamazepine also decreased active moiety concentrations. PMID- 17136450 TI - Further psychometric examination of the Tourette's Disorder Scales. AB - The Tourette's Disorder Scales (Shytle et al., 2003) are parent- (Tourette's Disorder Scales-Parent Rated; TODS-PR) and clinician-rated (Tourette's Disorder Scales-Clinician Rated; TODS-CR) measures that assess tics, obsessions, compulsions, inattention, hyperactivity, aggression, and emotional disturbances among children with tics. Although the TODS-PR/CR are being increasingly used in clinical trials, relatively little psychometric data have been reported. Subjects were 44 children and adolescents recruited in a university tic specialty clinic. Families were administered the TODS-CR and Yale Global Tic Severity Scale. Completion of the TODS-PR and Child Behavior Checklist were counterbalanced. Results indicated adequate to excellent internal consistency for the TODS-PR/CR scores. Excellent inter-rater agreement and convergent and divergent validity was found. These results provide further psychometric support for the TODS-PR and TODS-CR. PMID- 17136451 TI - Effects of a community toxic release on the psychological status of children. AB - This study sought to determine the emotional effects of a major community toxic release on children in the exposed community while controlling for the potential effects of response bias. Controlling for the response bias inherent in litigated contexts is an advance over previous studies of toxic exposure in children. A randomly selected representative sample of Exposed children (n = 31) was compared to a matched Control group (n = 28) from a nearby, unexposed community. Symptoms and complaints were assessed via interview with the children and their guardians, surveys and checklists, and well-established psychological instruments. Even when biased responding was controlled the Exposed children experienced more psychological distress, more physical symptoms, and greater general concern over their physical functioning than the Controls. The Exposed children also reported some concern about their future health and cancer risk but usually only if asked. Limitations and future research directions are discussed. PMID- 17136453 TI - Brief report: brain activation to social words in a sedated child with autism. AB - A functional magnetic resonance imaging (fMRI) study was performed on a 4-year old girl with autism. While sedated, she listened to three utterances (numbers, hello, her own first name) played through headphones. Based on analyses of the fMRI data, the amount of total brain activation varied with the content of the utterance. The greatest volume of overall activation was in response to numbers, followed by the word 'hello', with the least activation to her name. Frontal cortex activation was greatest in response to her name, with less activation for numbers, and the least for the word 'hello.' These findings indicate that fMRI can identify and quantify the brain regions that are activated in response to words in children with autism under sedation. PMID- 17136454 TI - Correlates of depression in the Korean American elderly: focusing on personal resources of social support. AB - Today's Korean American elderly are predominantly first-generation immigrants who face stern challenges of acculturation, which is often associated with depression. Social support has been identified as an effective personal resource for alleviating acculturative stress and achieving better mental health outcomes. The purposes of this study were to describe available sources of social support utilized by Korean elders and to examine the relationships among acculturative stress, social support, and depression. In particular, social support was operationalized as an integrative concept encompassing the size of the social network, satisfaction with the support received, and appraisals of the level of social support. This study was a secondary data analysis of an existing survey of 205 elderly Korean immigrants (>or=60 years) in a major metropolitan city on the East coast. Adult children were found to be the main source of support utilized by elders, even when the elder had a living spouse. Hierarchical multiple regression analysis revealed that higher acculturative stress and lower social support were associated with higher depression scores after demographics and health status were controlled for, whereas network size and satisfaction with support were not. Future interventions should address the cultural/social needs of these immigrants, not only by reinforcing their existing social network but also by providing additional support for their family members to prevent social isolation and depression in the population. PMID- 17136455 TI - Language barriers surrounding medication use among older Latinos. AB - Limited English language proficiency forms a significant challenge for many Latinos in clinical settings. Although medications are commonly used by older individuals as a means of maintaining good health and managing health problems, the extent to which English proficiency is related to medication use among older Latinos is not known. Focus groups were conducted with Latino, community-residing individuals aged 50 and over in eastern Massachusetts. Qualitative evaluation of the group interviews suggests that language is a barrier in dealing with medication for these individuals. Limited English proficiency appears to be related to feelings of being discriminated against in clinical and pharmacy settings. As well, communicating directly with health professionals in a common language is associated with level of trust and confidence in medical settings. Use of formal and informal interpreters, as well as seeking Spanish-speaking physicians and pharmacies with Spanish-speaking staff, are identified as strategies for overcoming health-related obstacles surrounding language. PMID- 17136456 TI - No association between p73 G4C14-to-A4T14 polymorphism and the risk of lung cancer in a Korean population. AB - A member of the p53 family, p73 may play an important role in the development of lung cancer. Variations in the DNA sequence in the p73 gene can lead to alterations in the production of p73 and/or activity, which can affect an individual's susceptibility to lung cancer. To test this hypothesis, this study examined the association between the G4C14-to-A4T14 polymorphism in the p73 gene and the risk of lung cancer in a Korean population. The p73 G4C14-to-A4T14 genotypes were determined in 582 lung cancer patients and 582 healthy age- and gender-matched control subjects. Compared with the GC/GC genotype, the GC/AT and the AT/AT genotypes were not significantly associated with the risk of lung cancer [adjusted odds ratio (OR) = 1.08, 95% confidence interval (CI) = 0.84 1.38; and adjusted OR = 1.37, 95% CI = 0.83-2.24, respectively]. In addition, the risk estimate for the combined variant genotype (GC/AT + AT/AT) was similar to that of the GC/GC genotype (a dominant model for the AT allele, adjusted OR = 1.12, 95% CI = 0.88-1.41). These results suggest that the p73 G4C14-to-A4T14 polymorphism does not significantly affect susceptibility to lung cancer in the Korean population. PMID- 17136457 TI - The effects of thematic importance on story recall among children with attention deficit hyperactivity disorder and comparison children. AB - This study examined the recall of televised stories for younger (4-6 years) and older (7-9 years) children with and without attention deficit hyperactivity disorder (ADHD) under two different viewing conditions (toys present/toys absent). Each child watched two Rugrats television programs, once with toys present and once with toys absent. Immediately after viewing a program, the child completed a free recall of the observed story. Comparison children's recall increased more than ADHD children's as importance level increased, and comparison children recalled more information overall than children with ADHD. When toys were present, children with ADHD retold less coherent stories than comparison children, as indexed by smaller correlations between the story units recalled and the order of these units in the story. In summary, children with ADHD demonstrated multiple difficulties in story comprehension. These findings add to our understanding of the differences in higher-order cognitive processing abilities between children with ADHD and comparison children, and suggest important areas of focus in designing more effective academic interventions for children with ADHD. PMID- 17136458 TI - Examining subtypes of behavior problems among 3-year-old children, Part III: investigating differences in parenting practices and parenting stress. AB - This study examined parenting of mothers and fathers of 3-year-old children who were classified as hyperactive (HYP), hyperactive and oppositional defiant (HYP/OD), and non-problem based on mothers' reports of behavior. Parents of children with HYP/OD were less warm, showed more negative affect, and more laxness than parents of non-problem children; however, contrary to prediction, they did not differ significantly from parents of children in the HYP group. Compared to mothers of non-problem children, mothers of children in the HYP/OD group reported significantly more parenting stress across all domains, whereas mothers of children in the HYP group only reported elevated levels of stress associated with having a difficult child. Theoretical and treatment implications are discussed. PMID- 17136459 TI - The effect of health compromising behaviors on preterm births. AB - OBJECTIVES: The objective of our study was to determine whether there were combined effects of smoking, alcohol, and illicit drug use during pregnancy on the frequency of preterm births, and if so, the magnitude of the association after adjusting for confounding factors. METHODS: We conducted a retrospective cohort study of singleton live births in Kansas City, Missouri from 1990-2002. We defined health compromising behaviors as the use of cigarettes, alcohol, and illicit drugs. The effect of these behaviors on preterm births was considered for each substance individually, and in combination. The rates of preterm births for these groups were calculated. Using logistic regression, adjusted odds ratios were used to estimate the relative risk of preterm births among these groups. RESULTS: Over 13% of infants born to women who smoked were preterm, compared to 9.6% for non-smokers. Of infants born to women who reported alcohol use, 17.3% were preterm compared to 10.1% for non-drinkers. Smoking and alcohol use in combination was associated with 18.0% preterm births, while alcohol and drug use in combination was associated with 20.8% preterm births. The use of all three substances was associated with 31.4% preterm births. CONCLUSION: Women who engaged in health compromising behaviors during pregnancy showed an increased proportion of preterm births compared to those who did not. There is significant interaction between these behaviors leading to higher rates of preterm births than predicted by their additive effects. To decrease preterm births, we must deal with the effects of smoking, drinking, and drug use simultaneously. PMID- 17136460 TI - Impact of prenatal care on infant survival in Bangladesh. AB - Despite improvements in public health in recent decades, levels of infant and child mortality remain unacceptably high, particularly in developing countries where primary healthcare services including prenatal care services are not universally available. Using information on 7,001 childbirths in five years preceding the 2004 Bangladesh Demographic and Health Survey, this study examined the relationship between receiving prenatal care during pregnancy and infant mortality using multivariate survival analysis. The results are presented in hazard ratios (HR) with 95% confidence intervals (95% CI). Results indicate that children of mothers who did not receive prenatal care during pregnancy were more than twice as likely to die during infancy as children whose mothers received prenatal care during pregnancy (HR=2.40, 95% CI: 1.74, 3.31) independent of child's sex, delivery assistance, birth order; mother's age at child birth, nutritional status, education level; household living conditions, and other factors. Children born to older mothers living in households without safe drinking water were at an increased risk. The study concludes that prenatal care is strongly negatively associated with infant mortality in Bangladesh independent of other risk factors. The results suggest that improving prenatal care services at the community level is key to improving child survival in Bangladesh. PMID- 17136461 TI - A controlled trial of web-based feedback for heavy drinking college students. AB - OBJECTIVE: Alcohol consumption has been a growing concern at U.S. colleges, particularly among first-year students, who are at increased risk for problems. This study tested the efficacy of the "electronic Check-Up to Go" (e-CHUG), a commercially-available internet program, at reducing drinking among a group of at risk college freshman. METHOD: The design was a randomized controlled trial: 106 freshmen students who reported heavy episodic drinking were randomly assigned to receive feedback or to assessment only. Assessment measures were completed at baseline, 8 weeks, and 16 weeks. RESULTS: At 8 weeks, the feedback group showed a significant decrease in drinks per week and peak BAC over control. By 16 weeks, the control group also declined to a point where there were no differences between groups. Changes in normative drinking estimates mediated the effect of the intervention. An additional 245 abstainers and light drinkers who were also randomized to condition did not show any intervention effect. CONCLUSIONS: This study provides preliminary support for the efficacy of this intervention at reducing short-term drinking among at-risk students. PMID- 17136462 TI - Preventing youth violence and delinquency through a universal school-based prevention approach. AB - Violence is an important public health problem among adolescents in the United States. Substance use and violence tend to co-occur among adolescents and appear to have similar etiologies. The present study examined the extent to which a comprehensive prevention approach targeting an array of individual-level risk and protective factors and previously found effective in preventing tobacco, alcohol, and illicit drug use is capable of decreasing violence and delinquency. Schools (N=41) were randomly assigned to intervention and control conditions. Participants in the 20 intervention schools received the Life Skills Training prevention program including material focusing on violence and the media, anger management, and conflict resolution skills. Survey data were collected from 4,858 sixth grade students prior to the intervention and three months later after the intervention. Findings showed significant reductions in violence and delinquency for intervention participants relative to controls. Stronger prevention effects were found for students who received at least half of the preventive intervention. These effects include less verbal and physical aggression, fighting, and delinquency. The results of this study indicate that a school-based prevention approach previously found to prevent tobacco, alcohol, and illicit drug use can also prevent violence and delinquency. PMID- 17136463 TI - Spectacular reproduction: Ron's Angels and mechanical reproduction in the age of ART (assisted reproductive technology). AB - Ron Harris captured the popular imagination in October 1999 with a website where he auctioned off the ova of fashion models to the highest bidder. This article treats the controversy surrounding Harris' site within a dual frame of critical theory's approach to reproduction and a folkloristic approach to discourse. The website fuses traditional narrative motifs and structures with the logic of advertising, seventies television, family-values rhetoric, and the fertility industry. I argue that the great attraction of ronsangels.com is that it put into relief the intervention of mechanical reproduction in human fertility together with the state of genetics at the turn of the 21st century. The result is not only a disconcerting aestheticization and commodification of biological reproduction, but also the biological reproduction of a particular aesthetic and moral code--a generation of reality by model. PMID- 17136464 TI - Lichen substances affect metal adsorption in Hypogymnia physodes. AB - Lichen substances are known to function as chelators of cations. We tested the hypothesis that lichen substances can control the uptake of toxic metals by adsorbing metal ions at cation exchange sites on cell walls. If true, this hypothesis would help to provide a mechanistic explanation for results of a recent study showing increased production of physodalic acid by thalli of the lichen Hypogymnia physodes transplanted to sites with heavy metal pollution. We treated cellulose filters known to mimic the cation exchange abilities of lichen thalli with four lichen substances produced by H. physodes (physodic acid, physodalic acid, protocetraric acid, and atranorin). Treated filters were exposed to solutions containing seven cations (Ca(2+), Cu(2+), Fe(2+), Fe(3+), Mg(2+), Mn(2+), and Na(+)), and changes to the solution concentrations were measured. Physodalic acid was most effective at influencing metal adsorption, as it increased the adsorption of Fe(3+), but reduced the adsorption of Cu(2+), Mn(2+), and Na(+), and to a lesser extent, that of Ca(2+) and Mg(2+). Reduced Na(+) adsorption matches with the known tolerance of this species to NaCl. The results may indicate a possible general role of lichen substances in metal homeostasis and pollution tolerance. PMID- 17136465 TI - Intonation and emotion in autistic spectrum disorders. AB - The classic picture of an autistic individual includes an impoverished ability to interpret or express emotion. The prosody of spoken language in autistic children is thought to lack emotional content. In this study, the verbal intonation of children with autism was examined and compared to that of children with Asperger Syndrome (AS) and normal controls (ctrl). Utterances elicited by repetition and by spontaneous story completion were analyzed by quantifying phonetic features (pitch, amplitude, and length) and comparing them to subjective ratings of produced emotion (happy, sad or angry). Since the most consistent phonetic correlate of these emotional targets has been demonstrated to be pitch range, speakers with autistic spectrum disorders were expected to have decreased pitch range; however in the repetition task, autistic subjects actually had a larger pitch range than the other groups. Other measures of intonation including amplitude, duration, and location of pitch peak revealed defects that are more complex than predicted. In spontaneous speech, autistic subjects performed more poorly on both phonetic targets and subjective ratings than ctrls, and AS subjects fell between autistics and normals. PMID- 17136466 TI - Antihyperlipidemic effect of Aronia melanocarpa fruit juice in rats fed a high cholesterol diet. AB - Aronia melanocrpa fruit juice (AMFJ) used in our experiment was very rich in phenolic substances (709.3 mg gallic acid equivalents/100 ml juice). Anthocyanins (106.8 mg cyanidin-3-glucoside equivalents/100 ml juice) were the main flavonoid group. The aim of this study was to assess the influence of AMFJ on plasma lipids and lipoprotein profile, and histopathology of liver and aorta in rats with dietary-induced hyperlipidemia. AMFJ was administered by gavage for 30 days at doses of 5, 10 and 20 ml/kg body weight to rats fed a standard diet (SD) or a 4% cholesterol-containing diet (4% ChD). The 4% ChD caused a significant elevation of plasma total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and triglycerides (TG). AMFJ did not significantly influence plasma lipids in rats fed the SD and significantly hindered the elevation of plasma TC, LDL-C and TG in rats fed the 4% ChD. High-density lipoprotein cholesterol (HDL-C) levels were not significantly influenced either by the 4% ChD or by AMFJ. Neither the cholesterol feeding, nor AMFJ treatment induced any histopathological changes in rat liver and aorta. In conclusion, AMFJ showed an antihyperlipidemic effect in rats with hyperlipidemia and could be valuable in reducing this factor of cardiovascular risk. PMID- 17136467 TI - Scalp topography and intracerebral sources for ERPs recorded during auditory target detection. AB - The goal of this study was to measure the scalp topography of the event-related potentials (ERPs) during the detection of improbable auditory targets and to determine the intracerebral sources of these potentials. ERPs were recorded when subjects listened to a sequence of spoken words and detected occasional (p = 0.2) targets defined either by the gender of the speaker (male/female) or the meaning of the word (animate/inanimate). Waveforms were evaluated in relation to category (target versus standard) and task (voice versus semantic). Dipole source analysis was performed using locations obtained from fMRI. Fronto-central negative waves (N450 and N600 ms) generated by sources in both the auditory cortex and frontal regions were larger for semantic tasks and larger over the left hemisphere. A large parieto-occipital positive wave (P700) occurring with a peak latency about 150 ms before the reaction time was mainly generated in the left temporo-parietal regions for the semantic task and bilaterally for the voice task. About 300 ms after the P700, a highly lateralized right temporo-parietal positive wave P1000r occurred with a source in the right temporo-parietal area. These results indicate three distinct physiological processes underlie the detection of auditory targets. Perceptual discrimination is related to interactions between the frontal and temporal regions, stimulus-response association occurs in the temporo parietal regions and post-perceptual processing in the right temporo-parietal region. PMID- 17136468 TI - Cortical activity prior to, and during, observation and execution of sequential finger movements. AB - The aim of this study was to provide further evidence for the existence of a mirror neuron system in humans using electroencephalography during the observation and execution of non-object-related movements. Event-related desynchronization and synchronization (ERD/ERS) were used to characterize brain activity prior to, and during, observation and execution of a finger movement in four frequency bands (7-10 Hz, 10-13 Hz, 13-20 Hz, and 20-30 Hz). Electroencephalograms (EEGs) were recorded from 19 electrode sites in eight participants. In all the frequency bands and electrode sites, results revealed that there was no significant differences in EEG cortical activity between the observation condition and the execution conditions. Comparison of the two stages of the movement (i.e., pre-movement and movement) in the observation and execution conditions showed, in most cases, that pre-movement ERD values were less than movement ERD values. Whilst there was not an identical match of EEG cortical indices, this study provides further support for the existence of a mirror neuron system in humans. The incomplete congruence may be explained by the different behaviors, the nature of the task and factors in the observed action coded by the mirror system. PMID- 17136469 TI - Pluripotency and chimera competence of an embryonic stem cell line from the sea perch (Lateolabrax japonicus). AB - A stable GFP-expressing (GFP(+)LJES1) cell strain was developed from the LJES1 cells obtained from sea perch (Lateolabrax japonicus,) embryos. GFP(+)LJES1 cells were induced in vitro by RA to differentiate into a variety of cell types and also had the ability to form embryoid body-like structures in suspension culture. To determine the differentiation potential of LJES1 cells in vivo, GFP(+)LJES1 cells were transplanted into sea perch and zebrafish embryos at mid-blastula stage. Twenty out of 478 transplanted sea perch embryos contained GFP-expressing LJES1 cells 24 h after microinjection. Fifteen chimera embryos developed into fry. In these chimeras, the GFP(+)LJES1 cells contributed to a variety of tissues including the head and trunk. In zebrafish, 221 embryos were microinjected with GFP(+)LJES1 cells and 22 chimera embryos and fries expressing GFP were obtained. Donor GFP(+)LJES1 cells contributed to various tissues in head and trunk of zebrafish embryos and hatched fry. PMID- 17136470 TI - Comparison of multiphase CT, FDG-PET and intra-operative ultrasound in patients with colorectal liver metastases selected for surgery. AB - BACKGROUND: For patients with colorectal liver metastases, resection is the treatment of choice. Careful selection of these patients is crucial in order to reduce the chance of unexpected findings at laparotomy and abandoning further surgical intervention. Here, we evaluate the predictive value of CT and FDG-PET of the liver and extrahepatic findings compared to findings during laparotomy and 6 months follow-up. METHODS: 131 consecutive patients, selected for hepatic surgery for colorectal liver metastases by CT and FDG-PET, were evaluated prospectively. During surgery, the liver was assessed by intra-operative ultrasound, palpation and histology. RESULTS: In 127 patients (97%), CT was true positive for liver metastases. In 3 patients, CT was false-positive and in 1 patient false-negative. In 126 patients (96%), FDG-PET was true-positive for liver metastases, in 2 patients FDG-PET was false-negative, in 3 patients true negative (negative FDG-PET, false-positive CT). At laparotomy a total of 363 liver metastases was identified: 63 lesions <10 mm [10 (16%) detected by both CT and FDG-PET], 172 lesions of 10-20 mm [123 (72%) CT-positive, 129 (75%) by FDG PET-positive], and 28 lesions >20 mm [124 (97%) CT-positive, 121 (95%) FDG-PET positive]. CT and FDG-PET missed approximately 30% of the smaller liver lesions, resulting in a significant change in clinical management during surgery in only nine patients. CONCLUSIONS: CT and FDG-PET have a similar diagnostic yield for the identification of liver metastases; both modalities being adequate on a patient-basis but inadequate to detect the smallest of liver lesions. However, the clinical relevance of the latter is limited. PMID- 17136471 TI - Histological features of melanoma sentinel lymph node metastases associated with status of the completion lymphadenectomy and rate of subsequent relapse. AB - BACKGROUND: The current recommendation for patients with cutaneous melanoma and a positive sentinel lymph node (SLN) biopsy is a completion lymph node dissection (CLND). This study sought to define a population of SLN-positive patients, based on their histological pattern of SLN metastases, who may not require CLND. METHODS: All patients with SLN-positive cutaneous melanoma who underwent CLND between March 1999 and December 2004 at a single academic institution were enrolled. Metastatic deposits in the SLN were categorized by their histological zone of involvement (subcapsular, parenchymal and/or sinusoidal). Logistic regression was used to examine the effect of SLN zone, size of nodal metastases, and other histological factors on CLND positivity. Kaplan-Meier and Cox models were used to study disease recurrence. RESULTS: A total of 127 patients were included, and 15.8% had positive non-sentinel nodes. In adjusted analyses, the size of the largest tumor deposit in the SLN was the only factor associated with CLND status. No patients with a tumor deposit 0.05). There were no changes for VO2-MAX (p>0.05) in either group. Results of this study indicate that beta-alanine supplementation delays the onset of neuromuscular fatigue (PWCFT) and the ventilatory threshold (VT) at submaximal workloads, and increase in TTE during maximal cycle ergometry performance. However, beta-alanine supplementation did not affect maximal aerobic power (VO2 MAX). In conclusion, beta-alanine supplementation appears to improve submaximal cycle ergometry performance and TTE in young women, perhaps as a result of an increased buffering capacity due to elevated muscle carnosine concentrations. PMID- 17136506 TI - Cell cycle checkpoints: the role and evaluation for early diagnosis of senescence, cardiovascular, cancer, and neurodegenerative diseases. AB - Maintenance of genomic integrity is critical for prevention of a wide variety of adverse cellular effects including apoptosis, cellular senescence, and malignant cell transformation. Under stress conditions and even during an unperturbed cell cycle, checkpoint proteins play the key role in genome maintenance by and mediating cellular response to DNA damage, and represent an essential part of the "cellular stress response proteome". Intact checkpoint signal transduction cascades check the presence of genome damage, trigger cell cycle arrest, and forward the information to the protein core of cell cycle machinery, replication apparatus, repair, and/or apoptotic protein cores. Genetic checkpoint defects lead to syndromes that demonstrate chromosomal instability, increased sensitivity to genotoxic stress, tissue degeneration, developmental retardation, premature aging, and cancer predisposition that is most extensively studied for the ATM checkpoint mutated in Ataxia telangiectasia. Tissue specific epigenetic control over the function of cell cycle checkpoints can be, further, misregulated by aberrant DNA methylation status. The consequent checkpoint dysregulation may result in tissue specific degenerative processes such as degeneration and calcification of heart aortic valves, diabetic cardiomyopathy, hyperhomocysteinemic cerebrovascular, peripheral vascular and coronary heart diseases, neurodegenerative disorders (Alzheimer and Parkinson diseases, amyotrophic lateral sclerosis, glaucoma), and accelerated aging frequently accompanied with cancer. This review focuses on the checkpoints shown to be crucial for unperturbed cell cycle regulation, dysregulation of which might be considered as a potential molecular marker for early diagnosis of and therapy efficiency in neurodegenerative, cardiovascular and cancer diseases. An application of the most potent detection technologies such as "Disease Proteomics and Transcriptomics" also considered here, allows a most specific selection of diagnostic markers. PMID- 17136507 TI - Synthesis of a versatile constrained analogue of dipeptide DG (Asp-Gly). AB - The synthesis of an orthogonally protected constrained analogue of dipeptide DG (Asp-Gly) is reported exploiting alkylation of a chiral lactam. The versatility of this analogue was proven by removal of t-Boc protecting group, followed by coupling under homogeneous conditions with t-Boc-Arg(Z(2))-Gly, to give a conformationally restricted analogue of RGDG tetrapeptide. PMID- 17136508 TI - Bio-available amino acids and mineral nitrogen forms in soil of moderately mown and abandoned mountain meadows. AB - The abandonment of traditional mowing methods of mountain meadows in the Czech Republic at the end of the last century has resulted in secondary re-colonization of these areas. Altered accumulation of plant biomass resulted in a deceleration of N turnover. A mountain meadow may be regarded as a N-limited ecosystem in which plant nutrition is dependent on direct uptake of soil amino acids. The composition and distribution of ammonium ions, nitrate ions and the 16 bio available proteinaceous amino acids were investigated in the top 7 cm of the Ah horizon of a Gleyic Luvisol in a long-term moderately mown meadow and an eleven year old, abandoned or uncut meadow. Ammonium N has a dominant role in both ecosystems. The moderately mown meadow showed accelerated N-turnover and higher net ammonization. The plant community showed a dependence on this form. Plant utilization of nitrates and amino acids appeared to be negligible. The uncut or abandoned meadow showed net ammonization from May (start of the experiment) through August, after which plant N-uptake consisted only of amino acids due to microbial immobilization. The release of bio-available nitrogen from spring until the beginning of summer in the Ah horizon was too low to explain total plant N uptake. Glutamic acid, arginine and aspartic acids had the highest concentrations of any of the amino acids analyzed. PMID- 17136509 TI - Using C' deviation to study structures of central amino acids in peptide fragments. AB - In this investigation, we attempted to study the backbone geometry of amino acids in peptides using C' deviation. Diameters of distribution were used to describe the various atomic structures, and scatter graphs provided visual evaluation. The length of peptide fragments and the secondary structure of amino acids in the central position of the peptide fragments were also analyzed. The results showed that the atomic distribution of the central amino acids of five-residue peptide fragments was much more restricted than that of their corresponding three-residue peptide fragments. In identical three-residue fragments, atoms of central amino acids with different secondary structures, were distributed in distinct areas. PMID- 17136510 TI - Applications and current challenges of proteomic approaches, focusing on two dimensional electrophoresis. AB - Since the formulation of the concept of "proteomics" in 1995, a plethora of proteomic technologies have been developed in order to study proteomes of tissues, cells and organelles. The powerful new technologies enabled by proteomic approaches have lead to the application of these methods to an exponentially increasing variety of biological questions for highly complex protein mixtures. Continuous technical optimization allows for an ever-increasing sensitivity of proteomic techniques. In this review, a brief overview of currently available proteomic techniques and their applications is given, followed by a more detailed description of advantages and technical challenges of two-dimensional electrophoresis (2-DE). Some solutions to circumvent currently encountered technical difficulties for 2-DE analyses are proposed. PMID- 17136511 TI - Quality and flavour stability of coffee substitute prepared by extrusion of wheat germ and chicory roots. AB - A mixture of roasted chicory roots and wheat germ (1:1 w/w) was subjected to extrusion processing for preparation of coffee substitute. Comparative studies concerning sensory characteristics and headspace volatiles were carried out between genuine coffee and a freshly prepared coffee substitute. The sensory evaluation revealed similarities between the two samples. The comparative odour profile analysis showed that the sweetish/caramel-like note scored higher in our coffee substitute sample than in real coffee, whereas the other odour quality attributes showed an opposite trend. The high quality of the fresh coffee substitute was correlated to the presence of volatiles that are responsible for the fresh coffee aroma, such as: 2-methylbutanal, 3-methylbutanal, 2-methylfuran and 2,3-butanedione in high concentration. Storage of coffee substitute samples revealed a noticeable decrease in concentration of the Strecker aldehydes and diketones and a remarkable increase in phenolic compounds, whereas pyrazine and furan derivatives showed no linear changes during storage. The ratio of 2,3 butanedione/2-methylfuran (B/M) was used as an indicator for aging of coffee substitute samples. The variation in this ratio (B/M) during storage for 6 months was consistent with that of the odour profile analysis. PMID- 17136512 TI - Regulation of apoptosis by protein S-nitrosylation. AB - S-nitrosylation/denitrosylation of critical cysteine residues on proteins serves as a redox switch that regulates the function of a wide array of proteins. A key signaling pathway that is regulated by S-nitrosylation is apoptotic cell death. Here we will review the proteins in apoptotic pathways that are known to be S nitrosylated by endogenous NO production. The targets and functional consequences of S-nitrosylation during apoptosis are multifaceted, allowing cells to fine tune their response to apoptotic signals. PMID- 17136513 TI - Increased egg estradiol concentration feminizes digit ratios of male pheasants (Phasianus colchicus). AB - The length ratio between individual digits differs between males and females in humans, other mammals, lizards, and one bird species. Sexual dimorphism in digit ratios and variation among individuals of the same sex may depend on differential exposure to androgens and estrogens during embryonic life. Organizational effects of sex hormones could cause the observed correlations between digit ratios and diverse phenotypic traits in humans. However, no study has investigated experimentally the effect of prenatal estrogens on digit ratios. We analyzed the effect of estradiol injection in ring-necked pheasant (Phasianus colchicus) eggs on digit ratios. Males from control eggs had higher ratios between the second or third and the fourth digit of the right foot compared to females. Estradiol treated eggs produced males with lower (feminized) right foot second to fourth digit ratio. Thus, we provided the first experimental evidence that prenatal exposure to physiologically high estrogen levels affects bird digit ratios. PMID- 17136514 TI - A new Early Oligocene peradectine marsupial (Mammalia)from the Burqin region of Xinjiang, China. AB - Tertiary marsupial records are very scarce in Asia. A new peradectine marsupial, Junggaroperadectes burqinensis gen. et sp. nov., is reported from the Early Oligocene Keziletuogayi Formation in the Burqin region, Xinjiang, China. This new species is based on a single right upper M2. The tooth possesses a straight centrocrista, a characteristic of peradectines. Its main cusps lean buccally, with the paracone being smaller and lower than the metacone. The conules and stylar cusps are weakly developed. These characters distinguish J. burqinensis from Euro-American Tertiary peradectines, but they also imply a close phylogenetic relationship to Siamoperadectes and Sinoperadectes, two Asian Early Miocene peradectines. PMID- 17136515 TI - Three QTLs for Botrytis cinerea resistance in tomato. AB - Tomato (Solanum lycopersicum) is susceptible to grey mold (Botrytis cinerea). Partial resistance to this fungus was identified in accessions of wild relatives of tomato such as S. habrochaites LYC4. In order to identify loci involved in quantitative resistance (QTLs) to B. cinerea, a population of 174 F(2) plants was made originating from a cross between S. lycopersicum cv. Moneymaker and S. habrochaites LYC4. The population was genotyped and tested for susceptibility to grey mold using a stem bioassay. Rbcq1, a QTL reducing lesion growth (LG) and Rbcq2, a QTL reducing disease incidence (DI) were identified. Rbcq1 is located on Chromosome 1 and explained 12% of the total phenotypic variation while Rbcq2 is located on Chromosome 2 and explained 15% of the total phenotypic variation. Both QTL effects were confirmed by assessing disease resistance in two BC(2)S(1) progenies segregating for either of the two QTLs. One additional QTL, Rbcq4 on Chromosome 4 reducing DI, was identified in one of the BC(2)S(1) progenies. F(2) individuals, homozygous for the Rbcq2 and Rbcq4 alleles of S. habrochaites showed a reduction of DI by 48%. QTLs from S. habrochaites LYC4 offer good perspectives for breeding B. cinerea resistant tomato cultivars. PMID- 17136516 TI - Clinical research in Italy in adult patients unable to consent: after implementation of the European Union's Directive 2001/20/CE. PMID- 17136517 TI - beta2 subunit-containing nicotinic receptors mediate the enhancing effect of nicotine on trace cued fear conditioning in C57BL/6 mice. AB - RATIONALE: Previous research indicates that acute nicotine administration enhances the acquisition of contextual fear conditioning and trace cued fear conditioning. Pharmacological inhibition of alpha4beta2 nicotinic acetylcholine receptors (nAChRs), but not alpha7 nAChRs, blocked the enhancing effect of nicotine on contextual fear conditioning. Similarly, genetic deletion of the beta2 nAChR subunit but not the alpha7 nAChR subunit blocked the enhancing effect of nicotine on contextual fear conditioning. OBJECTIVES: In the present study, nAChR subunit knockout mice were used to compare the involvement of beta2 subunit containing nAChRs and alpha7 subunit-containing nAChRs in the effects of nicotine on hippocampus-dependent trace cued fear conditioning and contextual fear conditioning. METHODS: beta2 nAChR subunit knockout mice, alpha7 nAChR subunit knockout mice, and their wild-type littermates received either nicotine or saline 5 minutes before training and testing. Mice were trained using five conditioned stimulus (CS; 30 s, 85 dB white noise)--trace (30 s)--unconditioned stimulus (US; 2 s footshock) pairings. Freezing to the context and freezing to the CS were assessed 24 h later. RESULTS: Both contextual and trace cued fear conditioning were enhanced by nicotine administration in wild-type littermates and in alpha7 nAChR subunit knockout mice. In contrast, neither contextual fear conditioning nor trace cued fear conditioning was enhanced by nicotine administration in beta2 nAChR subunit knockout mice. CONCLUSIONS: These results suggest that beta2 subunit-containing nAChRs but not alpha7 nAChR subunit-containing nAChRs are critically involved in the enhancing effect of nicotine on contextual and trace cued fear conditioning. PMID- 17136518 TI - Acute psychomotor, subjective and physiological responses to smoking in depressed outpatient smokers and matched controls. AB - RATIONALE: Cigarette smoking is highly prevalent in people diagnosed with depression, and depressed smokers are less likely to quit. Examining depressed smokers' responses to smoking will help determine the role of depression in maintaining cigarette smoking. OBJECTIVES: To determine the psychomotor, subjective and physiological effects of cigarette smoking in currently depressed smokers versus matched controls. MATERIALS AND METHODS: Fourteen currently depressed smokers and 14 never-depressed smokers, matched in age, gender, nicotine dependence and daily cigarette consumption, smoked three cigarettes at half-hourly intervals. All smokers were non-deprived. Self-reported mood and craving for cigarettes, performance on a simple reaction time task, expired-air carbon monoxide, heart rate and blood pressure were assessed before and after smoking each cigarette. Smoking topography was also assessed. RESULTS: Depressives and controls did not differ in terms of dependence on cigarettes or expired-air carbon monoxide. Topographic and cardiovascular measures were similar in depressed and control participants, suggesting that they smoke cigarettes in a similar manner. However, depressives displayed enhanced reaction time performance after the first cigarette. Positively reinforced craving was reduced after smoking each cigarette but returned to baseline levels within 30 min in depressed but not in control smokers. Depressed smokers also displayed higher levels of negatively reinforced craving. Both depressives and controls reported improved positive mood after smoking. CONCLUSIONS: Cigarette smoking in non-deprived depressed smokers enhances psychomotor performance and the reduction of positively reinforced craving in depressed smokers after smoking is transient, suggesting that enhanced craving may play a role in the maintenance of smoking in depression. PMID- 17136519 TI - Disposable microfluidic ELISA for the rapid determination of folic acid content in food products. AB - A micro-analytical system for rapid and quantitative analysis by inhibition immunoassay is presented and applied to the detection of folic acid. Eight polymer microchannels of 65-nL volume each and containing microelectrodes are embedded in a cartridge so that they can be operated simultaneously. All fluidic steps as well as the amperometric detection in the channels are operated by an instrument and software developed in-house. The fluidic steps of the immunoassay occur through hydrodynamic loading of the different solutions through the channels. The speed and duration of the flow and incubation parameters can thus be adapted to the biological and testing requirements. The effectiveness of the system was demonstrated by analysing folic acid concentrations in real infant formula samples within 5 min. In an effort to get a fully monitored assay, each fluidic step is monitored thanks to continuous amperometric detection of oxygen in the microchannel. PMID- 17136520 TI - A reexamination of the size-weight illusion induced by visual size cues. AB - The size-weight illusion induced by visually perceived sizes was reexamined to investigate whether this illusion is a sensory based or cognitive-based phenomenon. A computer-augmented environment was utilized to manipulate visual size information of target objects independently of their haptic information. Two physical cubes of equal mass (30.0 g) and size (3.0 x 3.0 x 3.0 cm) were suspended in parallel by wires attached to small graspable rings, in order to keep haptically obtained information constant between lifts. Instead of directly seeing each physical cube, subjects viewed 3D graphics of a cube with a wire and a ring that were precisely superimposed onto each physical cube. Seventeen subjects vertically lifted these augmented cubes, one after the other, by grasping the attached rings, and then reported their perception of cube heaviness. The graphical size of a comparison cube pseudo randomly varied for every comparison from 1.0 x 1.0 x 1.0 to 9.0 x 9.0 x 9.0 cm, while that of a standard cube remained constant (5.0 x 5.0 x 5.0 cm). Results indicated that the size-weight illusion frequently and systematically occurred for all the subjects such that when the comparison cube was relatively smaller than the standard cube, it was perceived to be heavier, and vice versa. As the size difference increased between the standard cube and the comparison cube, more subjects experienced the illusion, and vice versa. Follow-up tests showed occurrence of the size-weight illusion was significantly correlated with subject's sensitivity to discriminate weight, but not with sensitivity to discriminate visual size. Results suggest that the size-weight illusion induced by only visual size cues in an augmented environment is sensory based, and depends on an individual's integrated perception based on multimodal sensory information. PMID- 17136521 TI - Controlling posture using a plantar pressure-based, tongue-placed tactile biofeedback system. AB - The present paper introduces an original biofeedback system for improving human balance control, whose underlying principle consists in providing additional sensory information related to foot sole pressure distribution to the user through a tongue-placed tactile output device. To assess the effect of this biofeedback system on postural control during quiet standing, ten young healthy adults were asked to stand as immobile as possible with their eyes closed in two conditions of No-biofeedback and Biofeedback. Centre of foot pressure (CoP) displacements were recorded using a force platform. Results showed reduced CoP displacements in the Biofeedback relative to the No-biofeedback condition. The present findings evidenced the ability of the central nervous system to efficiently integrate an artificial plantar-based, tongue-placed tactile biofeedback for controlling control posture during quiet standing. PMID- 17136522 TI - Trial by trial effects in the antisaccade task. AB - The antisaccade task requires participants to inhibit the reflexive tendency to look at a sudden onset target and instead direct their gaze to the opposite hemifield. As such it provides a convenient tool with which to investigate the cognitive and neural systems that support goal-directed behaviour. Recent models of cognitive control suggest that antisaccade performance on a single trial should vary as a function of the outcome (correct antisaccade or erroneous prosaccade) of the previous trial. In addition, repetition priming effects suggest that the spatial location of the target on the previous trial may also influence current trial performance. Thus an analysis of contingency effects in antisaccade performance may provide new insights into the factors that influence the monitoring and modulation of the antisaccade task and other ongoing behaviours. Using a multilevel modelling analysis we explored previous trial effects on current trial performance in a large antisaccade dataset. We found (1) repetition priming effects following correct antisaccades; (2) contrary to models of cognitive control antisaccade error rates were increased on trials following an error, suggesting that failures to adequately maintain the task goal can persist across more than one trial; and (3) current trial latencies varied according to the previous trial outcome (correct antisaccade, slowly corrected error or rapidly corrected error). These results are discussed in terms of current models of antisaccade performance and cognitive control and further demonstrate the utility of multilevel modelling for analysing antisaccade data. PMID- 17136523 TI - Smooth pursuit eye movements in 1,087 men: effects of schizotypy, anxiety, and depression. AB - Individuals with schizotypal personality disorder or high scores in questionnaires measuring schizotypy are at high risk for the development of schizophrenia and they also share some of the same phenotypic characteristics such as eye-tracking dysfunction (ETD). The question arises whether these individuals form a distinct high-risk group in the general population or whether schizotypy and ETD co-vary in the general population with no distinct cutoff point for a high-risk group. A large sample of military conscripts aged 18-25 were screened using oculomotor, cognitive and psychometric tools for the purposes of a prospective study on predisposing factors for the development of psychosis. Schizotypy measured using the perceptual aberration scale (PAS) and the schizotypal personality questionnaire (SPQ), anxiety and depression, measured using the Symptom Checklist 90-R, had no effect on pursuit performance in the total sample. Small groups of individuals with very high scores in schizotypy questionnaires were then identified. These groups were not mutually exclusive. The high PAS group had higher root-mean-square error scores (a quantitative measure for pursuit quality) than the total sample, and the high disorganized factor of SPQ group had lower gain and higher saccade frequencies in pursuit than the total sample. The presence of significant differences in pursuit performance only for predefined high schizotypy groups favors the hypothesis that individuals with high schizotypy might present one or more high-risk groups, distinct from the general population, that are prone to ETD as that observed in schizophrenia. PMID- 17136524 TI - Cognitive load affects postural control in children. AB - Inferring relations between cognitive processes and postural control is a relatively topical challenge in developmental neurology. This study investigated the effect of a concurrent cognitive task on postural control in a sample of 50 nine-year-old children. Each subject completed two balance trials of 60 s, one with a concurrent cognitive task (cognitive load) and another with no cognitive load. The concurrent cognitive task consisted of mentally counting backwards in steps of 2. Twelve posturographic parameters (PPs) were extracted from the centre of pressure (CoP) trajectory obtained through a load cell force plate. Analysis of variance revealed significant differences in the majority of the extracted PPs. CoP was found to travel faster, farther, and with substantially different features demonstrating an overall broadening of the spectrum in the frequency domain. Nonlinear stability factors revealed significant differences when exposed to a concurrent cognitive task, showing an increase of instability in the intervention rate of the postural control system. By grouping children through selected items from Teachers Ratings and PANESS assessment, specific significant differences were also found both in time and frequency domain PPs, thus confirming the hypothesis of an interaction between cognitive processes (and their development), and postural control. PMID- 17136525 TI - Muscle activation and cutaneous reflex modulation during rhythmic and discrete arm tasks in orthopaedic shoulder instability. AB - In orthopaedic shoulder instability, muscle activity (EMG) is altered during unconstrained discrete arm movement tasks (e.g. elevation against a load). These findings have been ascribed to deficits in afferent feedback and neural control with glenohumeral instabilities resulting from orthopaedic injury. However, the integrity of neural control during shoulder movements in those with unstable shoulders is unclear. It is not known if there are altered EMG patterns during rhythmic arm movement or during discrete tasks involving no load, as would be experienced in many arm motions performed in daily living. The primary objective of this study was to evaluate neural control of arm movements between those with unstable shoulders and control participants, within a constrained arm movement paradigm involving both rhythmic arm cycling and discrete reaching. To achieve this objective, we determined if the amplitude and timing of EMG related to the movement pattern (background EMG) was significantly different between groups. Cutaneous reflexes were used to simulate a perturbation to the upper limb that would typically evoke a coordinated response. In the elevation phase of the movement path for anterior and posterior deltoid, upper trapezius, infraspinatus and serratus anterior, background EMG during rhythmic arm cycling was significantly (24%, p < 0.05) larger in unstable shoulders than in controls. No differences were found in background EMG between the groups during the discrete task. Significant differences (p < 0.05) were also noted in cutaneous reflexes between groups for both the rhythmic and discrete tasks with the reflex amplitudes being either increased or reduced in unstable shoulders as compared to controls. The differences in the background EMG and the cutaneous reflexes patterns in those with shoulder instabilities suggest that neural control is altered during rhythmic movement. PMID- 17136526 TI - A Bayesian model of the disambiguation of gravitoinertial force by visual cues. AB - The otoliths are stimulated in the same fashion by gravitational and inertial forces, so otolith signals are ambiguous indicators of self-orientation. The ambiguity can be resolved with added visual information indicating orientation and acceleration with respect to the earth. Here we present a Bayesian model of the statistically optimal combination of noisy vestibular and visual signals. Likelihoods associated with sensory measurements are represented in an orientation/acceleration space. The likelihood function associated with the otolith signal illustrates the ambiguity; there is no unique solution for self orientation or acceleration. Likelihood functions associated with other sensory signals can resolve this ambiguity. In addition, we propose two priors, each acting on a dimension in the orientation/acceleration space: the idiotropic prior and the no-acceleration prior. We conducted experiments using a motion platform and attached visual display to examine the influence of visual signals on the interpretation of the otolith signal. Subjects made pitch and acceleration judgments as the vestibular and visual signals were manipulated independently. Predictions of the model were confirmed: (1) visual signals affected the interpretation of the otolith signal, (2) less variable signals had more influence on perceived orientation and acceleration than more variable ones, and (3) combined estimates were more precise than single-cue estimates. We also show that the model can explain some well-known phenomena including the perception of upright in zero gravity, the Aubert effect, and the somatogravic illusion. PMID- 17136527 TI - Single retinal changing contrast (third) detector elicits NMDA receptor response and higher activity level of frog tectum neuron network. AB - The present study was designed to explore whether a discharge of a certain type of frog retinal ganglion cell [likely changing contrast (third) detector] can evoke NMDA response in frog tectum neurons and higher level of activity of tectal neuron network. Discharge of a single retinal ganglion cell was elicited by electrical stimulation of the retina. Evoked electrical activity of the tectum was recorded by the carbon-fiber microelectrode brought into the optic fiber layer F. We show that: (1) strong discharge of a frog individual retinal ganglion cell (third detector) has evoked NMDA response of tectal neurons and higher level of tectal neuron network activity characterized by prominent suprathreshold excitation of efferent neurons. Consequently, the firing of only one retinal ganglion cell (third detector) could lead to the activation of the tectobulbospinal tract and motor reaction. (2) The excitation of a retinotectal fiber of the first kind (axon of third detector) gave rise to the same effects as activation of a retinotectal fiber of the second kind (axon of fifth detector): the suprathreshold excitation of recurrent and efferent tectal neurons, the slow depolarizing potential (seen as the sNW), and the NMDA receptor activation were observed. However, stronger excitation (longer bursts of action potentials) was needed to evoke those effects in the considered case of the retinotectal input of the first kind. This difference could be attributed to the lower quantal size of neurotransmitter release in synapses of the retinotectal input of the first than second kind. PMID- 17136528 TI - Variability of quadriceps femoris motor neuron discharge and muscle force in human aging. AB - The purpose was to determine the contribution of visual feedback and the effect of aging on the variability of knee extensor (KE) muscle force and motor unit (MU) discharge. Single MUs were recorded during two types of isometric trials, (1) visual feedback provided (VIS) and then removed (NOVIS) during the trial (34 MUs from young, 32 from elderly), and (2) only NOVIS (66 MUs from young, 77 from elderly) during the trial. Recruitment threshold (RT) ranged from 0-37% MVC. Standard deviation (SD) and coefficient of variation (CV) of muscle force and MU interspike interval (ISI) was measured during steady contractions at target forces ranging from 0.3 to 54% MVC. Force drift (<0.5 Hz) was removed before analysis. VIS/NOVIS trials: the decrease in the CV of ISI from VIS to NOVIS was greater for MUs from elderly (12.5 +/- 4.1 to 9.94 +/- 2.6%) than young (10.6 +/- 3.3 to 10.3 +/- 2.8%, age group x vision interaction, P = 0.006). The change in CV of force from VIS to NOVIS was significantly greater for elderly (1.45 to 1.05%) than young (1.42 to 1.41%). NOVIS only trials: for all MUs, the average RT (6.6 +/- 7.7 % MVC), target force above RT (1.20 +/- 2.7% MVC), SD of ISI (0.012 +/- 0.005 s), and CV of ISI (11.1 +/- 3.3%) were similar for young and elderly MUs. The CV of force was similar between age groups for trials between 0 and 3% MVC (1.74 +/- 0.74%) and was greater for young subjects from 3 to 10% MVC (1.47 +/- 0.5 vs. 1.21 +/- 0.4%) and >10% MVC (1.44 +/- 0.6 vs. 1.01 +/- 0.3%). The CV of ISI was similar between age groups for MUs in 0-3, 3-10, and >10% bins of RT. Thus, the contribution of visuomotor correction to the variability of motor unit discharge and force is greater for elderly adults. The presence of visual feedback appears to be necessary to find greater discharge variability in motor units from the knee extensors of elderly adults. PMID- 17136529 TI - Visual-vestibular influences on locomotor adjustments for stepping over an obstacle. AB - Combined visual and vestibular influences on locomotor control, particularly in changing environments, are little understood. We studied such influences on body orientation and foot trajectory control during level walking and obstacle avoidance. Six young adults walked on the level and over an obstacle while vision was present or occluded as well as while vestibular information was intact or perturbed using galvanic vestibular stimulation (GVS). The occlusion of vision caused a slowing of gait during obstacle avoidance as well as increased clearance of the leading limb over the obstruction. GVS caused lateral deviations in head and trunk roll angles as well as in foot and trunk displacements, but these lateral deviations were the same during both level walking and obstacle avoidance. In addition, GVS had no effect at all on sagittal plane factors such as speed, foot proximity to the obstacle and vertical clearance over the obstacle. Overall, there is a complex visual control of bilateral obstacle avoidance, but the lack of differences in GVS effects between level and obstructed walking shows that vestibular information is not upregulated for obstacle avoidance. In addition, the robust indifference of anterior foot placement and body displacement to significant lateral deviations from GVS suggests an orthogonally based sensori-locomotor control. PMID- 17136530 TI - Modulation of neuronal activity in CNS pain pathways following propofol administration in rats: Fos and EEG analysis. AB - We studied Fos expression in the central nociceptive pathways at different sedative levels in order to clarify the central mechanism of propofol's nociceptive action. Sprague-Dawley rats received propofol (PRO) or pentobarbital (PEN) and were divided into two groups with different doses of drug administration (light and deep sedative levels) based on the electroencephalogram analysis. Rats at each sedative level received heat stimulation to their face and Fos immunohistochemistry was performed at various brain sites. We also infused lidocaine into the jugular vein to test whether PRO directly activated nociceptors distributed in the vein. Fos expression in two major ascending pain pathways (lateral and medial systems) and descending modulatory system were precisely analyzed following intravenous (i.v.) administration of PRO or PEN. Many Fos protein-like immunoreactive (Fos protein-LI) cells were expressed in the trigeminal spinal nucleus caudalis (Vc), parabrachial nucleus, parafascicular nucleus, a wide area of the primary somatosensory cortex, anterior cingulate cortex, amygdala, periaqueductal gray, solitary tract nucleus, and lateral hypothalamus following heating of the face during PRO or PEN infusion. The number of Fos protein-LI cells was significantly greater in many Central nervous system regions during PRO infusion compared with PEN. Fos expression was significantly greater in the Vc and Periaqueductal gray following greater amount of PRO infusions compared, whereas they were significantly smaller in the Vc in the rats with PEN infusion. The Fos expression was significantly depressed following i.v. infusion of lidocaine before PRO administration. The present findings suggest that PRO is involved in the enhancement of Vc activity through direct activation of the primary afferent fibers innervating veins, resulting in pain induction during infusion. PMID- 17136531 TI - Is the human masticatory system devoid of recurrent inhibition? AB - The aim of the present study was to investigate the existence or otherwise of a functional recurrent inhibitory system (Renshaw cell system) in the motoneurons that innervate human masticatory muscles. In a previous study, L: acetylcarnitine (L: -Ac), a substance known to potentiate recurrent inhibition in humans was found to alter, in a specific way, the discharge variability, and the synchronous activity of motor units depending on the presence or absence of recurrent inhibition in the corresponding motoneuron pool. Using a similar paradigm, we have recorded the tonic discharge activity of motor unit pairs from the masseter muscle during voluntary isometric contraction while subjects were undergoing continuous intravenous saline (SAL, NaCl 0.9%) perfusion. Following a brief baseline-recording period, the subjects were given a test injection of either L: -Ac or isotonic saline (SAL) in a double blind manner. The variability, synchronization, and coherence between the motor unit discharges were analysed during three successive periods: pre-injection, during injection, and post injection, each lasting 2-3 min. Neither L: -Ac nor SAL injection induced a significant change in the inter-spike interval (ISI) or the coefficient of variation of the ISIs in the motor units tested. There were also no significant changes in the pattern of synchronous activity or in the coherence, which reflects the common frequency content of the unit discharges. Reminiscent of what had been observed previously with motoneurons without recurrent inhibition in the Abductor Digitorum Minimi muscle, the lack of effects of L: -Ac injection on the firing behaviour of masseter motoneurons may suggest that classical Renshaw cell inhibition is lacking in this motoneuron pool. PMID- 17136532 TI - Altered cortical integration of dual somatosensory input following the cessation of a 20 min period of repetitive muscle activity. AB - The adult human central nervous system (CNS) retains its ability to reorganize itself in response to altered afferent input. Intracortical inhibition is thought to play an important role in central motor reorganization. However, the mechanisms responsible for altered cortical sensory maps remain more elusive. The aim of the current study was to investigate changes in the intrinsic inhibitory interactions within the somatosensory system subsequent to a period of repetitive contractions. To achieve this, the dual peripheral nerve stimulation somatosensory evoked potential (SEP) ratio technique was utilized in 14 subjects. SEPs were recorded following median and ulnar nerve stimulation at the wrist (1 ms square wave pulse, 2.47 Hz, 1x motor threshold). SEP ratios were calculated for the N9, N11, N13, P14-18, N20-P25 and P22-N30 peak complexes from SEP amplitudes obtained from simultaneous median and ulnar (MU) stimulation divided by the arithmetic sum of SEPs obtained from individual stimulation of the median (M) and ulnar (U) nerves. There was a significant increase in the MU/M + U ratio for both cortical SEP components following the 20 min repetitive contraction task, i.e. the N20-P25 complex, and the P22-N30 SEP complex. These cortical ratio changes appear to be due to a reduced ability to suppress the dual input, as there was also a significant increase in the amplitude of the MU recordings for the same two cortical SEP peaks (N20-P25 and P22-N30) following the typing task. No changes were observed following a control intervention. The N20 (S1) changes may reflect the mechanism responsible for altering the boundaries of cortical sensory maps, changing the way the CNS perceives and processes information from adjacent body parts. The N30 changes may be related to the intracortical inhibitory changes shown previously with both single and paired pulse TMS. These findings may have implications for understanding the role of the cortex in the initiation of overuse injuries. PMID- 17136533 TI - Glutamatergic systems in the bed nucleus of the stria terminalis, effects on cardiovascular system. AB - The bed nucleus of the stria terminalis (BST) is a part of the limbic system. Two studies have shown that microinjection of L: -glutamate in the BST elicited cardiovascular depressive and bradycardic responses, but in one study, both pressor and depressor responses were observed in the chemical stimulation of BST by glutamate in the urethane-anesthetized rats. Also, the roles of glutamate receptor subtypes have not been investigated yet. The aim of this study was to find the effects of glutamate and its receptors on the blood pressure and heart rate in the BST of urethane-anesthetized rats. The drugs (50 nl) were microinjected into the BST of anaesthetized rats. The blood pressure and heart rate were recorded throughout each experiment. The average changes in the mean arterial pressure and heart rate at different intervals were compared both within each case group and between the case and the control groups, using repeated measures ANOVA. Microinjection of L: -glutamate (0.25 M) into the BST resulted in the decrease of the mean arterial pressure (-18.85 +/- 3.84 mmHg) and heart rate (-18 +/- 4 beats/min). Injection of AP5, antagonist of glutamate NMDA receptor (2.5 , 5 mM) and CNQX, antagonist of glutamate AMPA receptor (0.5, 1 mM) had no significant effect on the mean arterial pressure and heart rate. Either Ap5 or CNQX, when co-injected with glutamate, abolished the depressor and bradycardic effects of glutamate, suggesting that simultaneous activation of both glutamate receptors is necessary for the effect of glutamate system to emerge. PMID- 17136534 TI - Reversible splenial abnormality in hypoglycemic encephalopathy. AB - INTRODUCTION: Lesions involving the splenium of the corpus callosum (SCC) have been rarely reported in cases of hypoglycemic brain injury. METHODS: We identified signal abnormalities in the SCC in three adult patients with hypoglycemic encephalopathy by using diffusion-weighted imaging (DWI) on a 1.5-T MR scanner. Repeat DWI was performed in all patients following a marked clinical improvement, and MR angiography and routine MRI were also performed. We examined each patient's detailed medical history and blood laboratory tests in order to exclude other conditions causing similar SCC abnormalities. RESULTS: Initial DWI was performed during which each patient showed altered mental status that was attributed to profound hypoglycemia. We observed an identical pattern of DWI abnormality characterized by high signals in the SCC with apparent diffusion coefficient reductions that were reversed completely within several days following appropriate correction of hypoglycemia. T2-weighted or FLAIR images also showed no residual lesion in the SCC and MR angiography was normal in all patients. CONCLUSION: These case reports suggest that the SCC should be added to the list of selective vulnerability to hypoglycemia and that hypoglycemia, in turn, be included in the differential diagnosis of reversible SCC abnormalities. PMID- 17136535 TI - Influence of metal ionic characteristics on their biosorption capacity by Saccharomyces cerevisiae. AB - The influence of metal ionic characteristics on their biosorption capacity was analyzed using quantitative structure-activity relationships model. The waste biomass of Saccharomyces cerevisiae was used as biosorbent to adsorb 10 kinds of metal ions, and their maximum biosorption capacity (q (max)) was determined by the Langmuir isotherm model. The values of q (max) decreased in the following order (in millimole per gram): Pb(2+) (0.413) > Ag(+) (0.385) > Cr(3+) (0.247) > Cu(2+) (0.161) > Zn(2+) (0.148) > Cd(2+) (0.137) > Co(2+) (0.128) > Sr(2+) (0.114) > Ni(2+) (0.108) > Cs(+) (0.092). Twenty-two parameters of physiochemical characteristics of metal ions were selected and correlated with q (max), i.e., OX, AN, r (Angstroms), DeltaIP (eV), DeltaE (0) (V), X (m), |log K (OH)|, X(m)(2)(r), Z*(2)/r, AN/DeltaIP, sigma rho, AR, AW, IP, AR/AW, Z/r (2), Z/AR(2), Z/r, Z/AR, Z*(2)/r., Z*, N. The linear regression analysis showed that the covalent index [Formula: see text] was correlated well with q (max) for all metal ions tested in the following equation: q (max) = 0.029 + 0.061 (X(m)(2)r) (R (2) = 0.70). It suggested that the greater the covalent index value of metal ion was, the greater the potential to form covalent bonds with biological ligands, such as sulphydryl, amino, carboxyl, hydroxyl groups, etc. on the biomass surface, and the higher the metal ion biosorption capacity was. Classification of metal ions, for divalent ion or for soft-hard ion could improve the linear relationship (R (2) = 0.89). The equation could be used to predict the biosorption capacity of metal ions. PMID- 17136536 TI - Steroid-1-dehydrogenase of Mycobacterium sp. VKM Ac-1817D strain producing 9alpha hydroxy-androst-4-ene-3,17-dione from sitosterol. AB - The strain of Mycobacterium sp. VKM Ac-1817D forms 9alpha-hydroxy-androst-4-ene 3,17-dione (9-OH-AD) as a major product from sitosterol. The formation of 9-OH-AD was accompanied with its partial destruction due to residual steroid-1 dehydrogenase (St1DH) activity. The activity was found to be induced by androst-4 ene-3,17-dione (AD), while other intermediates of sitosterol oxidation did not influence 1(2)-dehydrogenation. The enzyme is located mainly in the cytosolic fraction. The cytosolic St1DH (dimer, M (r) approximately 58 kDa) was partially purified by ammonium sulfate fractionation, ion-exchange chromatography on DEAE Sepharose and Phenyl-Sepharose, and gel filtration on Bio-Gel A-0.5M. It expressed the St1DH activity toward both AD and 9-OH-AD. PMID- 17136537 TI - New insights on toluene biodegradation by Pseudomonas putida F1: influence of pollutant concentration and excreted metabolites. AB - The influence of toluene concentration on the specific growth rate, cellular yield, specific CO(2), and metabolite production by Pseudomonas putida F1 (PpF1) was investigated. Both cellular yield and specific CO(2) production remained constant at 1.0 +/- 0.1 g biomass dry weight (DW) g(-1) toluene and 1.91 +/- 0.31 g CO(2) g(-1) biomass, respectively, under the tested range of concentrations (2 250 mg toluene l(-1)). The specific growth rate increased up to 70 mg toluene l( 1). Further increases in toluene concentration inhibited PpF1 growth, although inhibitory concentrations were far from the application range of biological treatment processes. The specific ATP content increased with toluene concentration up to toluene concentrations of 170 mg l(-1). 3-Methyl catechol (3 MC) was never detected in the cultivation medium despite being an intermediary in the TOD pathway. This suggested that the transformation from toluene to 3-MC was the limiting step in the biodegradation process. On the other hand, benzyl alcohol (BA) was produced from toluene in a side chain reaction. This is, to the best of our knowledge, the first reported case of methyl monoxygenation of toluene by PpF1 not harboring the pWW0 TOL plasmid. In addition, the influence of 3-MC, BA, and o-cresol on toluene degradation was investigated respirometrically, showing that toluene-associated respiration was not significantly inhibited in the presence of 10-100 mg l(-1) of the above-mentioned compounds. PMID- 17136538 TI - Study of the oxygen transfer in a disposable flexible bioreactor with surface aeration in vibrated medium. AB - The oxygen mass transfer is a critical design parameter for most bioreactors. It can be described and analyzed by means of the volumetric mass transfer coefficient K(L)a. This coefficient is affected by many factors such as geometrical and operational characteristics of the vessels, type, media composition, rheology and microorganism's morphology and concentration. In this study, we aim to develop and characterize a new culture system based on the surface aeration of a flexible, single-used bioreactor fixed on a vibrating table. In this context, the K(L)a was evaluated using a large domain of operating variables such as vibration frequency of the table, overpressure inside the pouch and viscosity of the liquid. A novel method for K(L)a determination based on the equilibrium state between oxygen uptake rate and oxygen transfer rate of the system at given conditions was also developed using resting cells of baker's fresh yeast with a measured oxygen uptake rate of 21 mg g(-1) h(-1) (at 30 degrees C). The effect of the vibration frequency on the oxygen transfer performance was studied for frequencies ranging from 15 to 30 Hz, and a maximal K(L)a of 80 h(-1) was recorded at 30 Hz. A rheological study of the medium added with carboxymethylcellulose at different concentrations and the effect of the liquid viscosity on K(L)a were determined. Finally, the mixing time of the system was also measured using the pH method. PMID- 17136539 TI - Nuclear medicine in the treatment of neuroendocrine tumours--problems and perspectives. PMID- 17136541 TI - Prolonged molecular remission in a newly diagnosed acute promyelocytic leukaemia with a severe cardiomyopathy using low-dose gemtuzumab ozogamicin and all-trans retinoic acid. PMID- 17136542 TI - Phase I study of TZT-1027, a novel synthetic dolastatin 10 derivative and inhibitor of tubulin polymerization, which was administered to patients with advanced solid tumors on days 1 and 8 in 3-week courses. AB - PURPOSE: To determine the maximum tolerated dose (MTD), dose-limiting toxicity (DLT), and pharmacokinetics of TZT-1027 (soblidotin), a dolastatin 10 analogue, in Japanese patients with advanced solid tumors when administered on days 1 and 8 in 3-week courses. METHODS: Eligible patients had advanced solid tumors that failed to respond to standard therapy or for which no standard therapy was available, and also met the following criteria: prior chemotherapy < or = 2 regimens, Eastern Cooperative Oncology Group (ECOG) performance status < or = 1, and acceptable organ function. The MTD was defined as the highest dose at which no more than one of six patients experienced a DLT during course 1. Pharmacokinetic samples were collected in courses 1 and 2. RESULTS: Eighteen patients were enrolled in the present study. Three doses (1.5, 1.65, and 1.8 mg/m(2)) were evaluated. Neutropenia was the principal DLT at doses of 1.65 and 1.8 mg/m(2). In addition, one patient also experienced grade 3 pneumonia with neutropenia, and another patient experienced grade 3 constipation, neuropathy, grade 4 neutropenia, and hyponatremia as DLTs at 1.65 mg/m(2). Phlebitis, the most frequent nonhematological toxicity, was improved by administration of additional saline after TZT-1027 administration. The MTD was 1.5 mg/m(2), at which DLT was not observed in a total of nine patients. The pharmacokinetic profile did not differ from that for the European population. One patient with metastatic esophageal cancer achieved partial response, and each of two patients with non-small cell lung cancer had a minor response. CONCLUSIONS: When TZT-1027 was administered on days 1 and 8 in 3-week courses to Japanese patients, the MTD was 1.5 mg/m(2) and was lower than the value of 2.4 mg/m(2) in European patients. However, antitumor activity was observed at low doses. TZT-1027 was tolerated well at the MTD, without grade 3 nonhematological toxicities or neutropenia up to grade 2. TZT-1027 is a promising new tubulin polymerization inhibitor that requires further investigation in phase II studies. PMID- 17136543 TI - Sunitinib malate. AB - Recently, there has been a growing interest in understanding the role of receptor tyrosine kinases (RTK), such as vascular endothelial growth factor receptor (VEGFR), platelet-derived growth factor receptor (PDGFR), stem cell factor receptor (KIT), and fms-like tyrosine kinase 3 (FLT3), in promoting tumor angiogenesis, tumor growth and metastasis. Sunitinib (sunitinib malate; SU11248; SUTENT; Pfizer Inc, New York, NY, USA) is a novel, orally bio-available, oxindole, multi-targeted tyrosine kinase inhibitor with high binding affinity for VEGFR and PDGFR which has shown anti-tumor and anti-angiogenic activities. This drug recently received approval from the US Food and Administration (FDA) in two indications simultaneously: advanced renal cell carcinoma (adRCC) and gastrointestinal stromal tumors (GIST), in patients who are resistant or intolerant to the treatment with imatinib. The present article reviews the recent pharmacologic and clinical data related to the use of this new promising drug in the field of oncology. PMID- 17136544 TI - Spontaneous pneumomediastinum and subcutaneous emphysema in systemic sclerosis. AB - A 40-year-old woman with known systemic sclerosis presented with dyspnea. She had been treated for pneumonia, 2 months prior to the present admission and at this time presented with sudden dyspnea and was found to have a spontaneous pneumomediastinum and subcutaneous emphysema. Pneumomediastinum is an extremely rare complication in patients with systemic sclerosis. PMID- 17136545 TI - Regeneration of roots from callus reveals stability of the developmental program for determinate root growth in Sonoran Desert Cactaceae. AB - In some Sonoran Desert Cactaceae the primary root has a determinate root growth: the cells of the root apical meristem undergo only a few cell division cycles and then differentiate. The determinate growth of primary roots in Cactaceae was found in plants cultivated under various growth conditions, and could not be reverted by any treatment tested. The mechanisms involved in root meristem maintenance and determinate root growth in plants remain poorly understood. In this study, we have shown that roots regenerated from the callus of two Cactaceae species, Stenocereus gummosus and Ferocactus peninsulae, have a determinate growth pattern, similar to that of the primary root. To demonstrate this, a protocol for root regeneration from callus was established. The determinate growth pattern of roots regenerated from callus suggests that the program of root development is very stable in these species. These findings will permit future analysis of the role of certain Cactaceae genes in the determinate pattern of root growth via the regeneration of transgenic roots from transformed calli. PMID- 17136546 TI - Absence of circadian and photoperiodic conservation of energy expenditure in three rodent species. AB - According to a traditional homeostatic view, living beings spend metabolic energy at a constant rate, just like a light bulb spends electrical energy, so that energy expenditure can be expressed in units of watts. However, research conducted during the last half-century has evinced pronounced circadian variation in physiological processes, not only demonstrating circadian rhythmicity in energy expenditure but also raising the hypothesis that energy expenditure may be regulated on a daily (circadian) basis rather than on a constant-rate (homeostatic) basis. In the present study, the hypothesis of circadian (and photoperiodic) conservation of energy expenditure was tested in three rodent species: domestic mice, Nile grass rats, and Syrian hamsters. Two correlates of energy expenditure (running-wheel activity and food intake) and a classic index (oxygen consumption) were used. Changes in energy expenditure were studied in animals maintained under light-dark cycles (LDs) with periods shorter or longer than 24 h as well as in animals maintained under 24-h LDs with short and long photophases. In none of the conditions in any of the species was evidence found in support of the hypothesis of circadian (or photoperiodic) conservation of energy expenditure. Energy expenditure was generally conserved on a homeostatic basis. PMID- 17136547 TI - Evidence for a differential expression of fibronectin splice forms ED-A and ED-B in Crohn's disease (CD) mucosa. AB - BACKGROUND AND AIMS: Fibronectin (FN) is an essential factor for the induction of migration of primary colonic lamina propria fibroblasts (CLPF). The FN isoform ED A is an important inducer of migration. Recently, we have shown that CLPF isolated from inflamed Crohn's disease (CD) mucosa migrated significantly less than control CLPF. We, therefore, investigated changes in FN or integrin expression that could be relevant for CLPF migration. MATERIALS AND METHODS: mRNA of control-CLPF and CLPF isolated from fibrotic mucosa of CD patients was subtractively hybridized. Expression of FN, ED-A, and ED-B in frozen sections from intestinal mucosa was determined by immunohistochemistry. The mRNA expression of the FN isoforms in control, CD, and fibrosis biopsies was quantified by real-time polymerase chain reaction (PCR). Integrin alpha5beta1 protein and mRNA expression was analyzed by fluorescence activated cell sorting (FACS) and PCR, respectively. RESULTS: Subtractive hybridization indicated differential regulation of FN isoform expression in CD. The immunohistochemical analysis of FN protein revealed a reduction of FN isoforms in inflamed CD mucosa compared to control mucosa. In CD fistulae, the ED-A and ED-B isoforms were virtually absent. In fibrotic mucosa, both proteins were increased. Real-time PCR showed a decrease of FN and ED-A expression during mucosal inflammation in CD in contrast to UC and a significant increase of FN and isoforms in CD fibrosis. No difference was found for protein and mRNA of integrin alpha5beta1 in control, CD, and fibrosis CLPF by FACS and PCR. CONCLUSION: Downregulated expression of migration-inducing FN-isoforms in contrast to unchanged FN receptor expression may contribute to the observed alterations of CD CLPF migration. PMID- 17136548 TI - TPPP/p25 in brain tumours: expression in non-neoplastic oligodendrocytes but not in oligodendroglioma cells. PMID- 17136549 TI - Notch1 and its ligand Jagged1 are present in remyelination in a T-cell- and antibody-mediated model of inflammatory demyelination. AB - The Notch receptor and its ligands are involved in myelination in central nervous system (CNS) development. Re-expression of this pathway in the adult CNS has been proposed to hamper remyelination in multiple sclerosis. Previous studies also revealed that pharmacological inhibition of Notch signaling ameliorates experimental autoimmune encephalomyelitis (EAE). However, in a recent study in toxin-induced demyelination constituents of the Notch signaling pathway were demonstrated in remyelinating lesions indicating that remyelination may occur in the presence of Notch signaling. We examined the expression of Notch1 immunoreactivity (IR) and Jagged1-IR in EAE induced by myelin-oligodendrocyte glycoprotein (MOG). In this model, the combined action of T cells, antibodies and the complement cascade yields a pathology closely reflecting multiple sclerosis. Notch1 and its ligand Jagged1 were differentially expressed in the lesions of MOG EAE. Notch1-IR on macrophages was highest in actively demyelinating and lowest in remyelinating lesions. The amount of Notch1-positive astrocytes increased during the lesion evolution from demyelination to remyelination. Notch1-positive oligodendrocytes were exclusively present in remyelinating lesions and not found in lesions without signs of remyelination. Astrocytes represented the major source of Jagged1-IR in demyelination and remyelination. In conclusion, our study proves that constituents of the Notch pathway are expressed in remyelination in an animal model of T-cell- and antibody-mediated CNS demyelination. Thus, it is unlikely, at least in the paradigm of MOG-EAE, that Notch signaling is responsible for a failure of remyelination. PMID- 17136550 TI - Prescribing patterns of anti-emetic drugs during pregnancy in Germany. AB - OBJECTIVES: The purpose of this study was to examine the prescribing patterns of anti-emetics for pregnant women in Germany. METHOD: A large, nation-wide German statutory sickness fund provided data of reimbursed prescriptions and personal data for all insured women giving birth between June 2000 and May 2001 (n = 41,293). The prescribed drugs were classified according to the Anatomical Therapeutic Chemical code. RESULT: About 13.9% (n = 5,746) of all pregnant women received an anti-emetic prescription. Over 98.5% of these women were treated with anti-emetics that are considered safe. But only 27.8% received the recommended anti-histamine meclozine and 2.1% the safest drug pyridoxine. CONCLUSION: As there are no official guidelines for the treatment of nausea and vomiting during pregnancy in Germany, many different drugs are used, sometimes ignoring effectiveness and even safety. Therefore, evidence-based guidelines for anti emetic therapy are needed. PMID- 17136551 TI - Poland's syndrome and head-and-neck tumour: an unusual association causing a reconstruction dilemma. AB - Poland's syndrome is a rare congenital anomaly characterized by unilateral chest wall hypoplasia and ipsilateral hand abnormalities. The literature data suggest its sporadic nature. The prevailing theory concerning its cause is hypoplasia of the subclavian artery or its branches, which may lead to a range of developmental changes. Relationships have been demonstrated between tumours and Poland's syndrome and also between tumours and other developmental defects. The explanation may lie in abnormal homeobox and tumour suppressor genes. This paper presents the first literature report of a malignant tonsillo-lingual tumour with metastatic neck involvement in a patient with partial Poland's sequence. In consequence of the aplasia of the pectoralis major muscle, an alternative (a free radical forearm flap) to the routine head-and-neck reconstruction (pedicled pectoralis major flap) was necessitated following tumour excision and radical neck dissection. This case report surveys the diagnostic and therapeutic considerations when previously unnoticed Poland's syndrome is diagnosed in a patient with head-and-neck cancer. One year following major head-and-neck surgery, our patient is tumour-free. PMID- 17136552 TI - A case of Moebius syndrome presenting with congenital bilateral vocal cord paralysis. AB - We describe a female infant with bilateral facial paralysis and abducens palsy. To the best of our knowledge, this is the first report of Moebius syndrome presenting with congenital bilateral vocal cord paralysis (CBVCP). Although CBVCP can be part of a recognizable syndrome, i.e. Down syndrome, 22q deletion syndrome, Robinow's syndrome and cerebro-oculo-facio-skeletal syndrome, no reports of Moebius syndrome with CBVCP were found in the literature. CBVCP is often associated with central nervous system abnormalities. However, our patient had no detectable brain abnormalities. The etiology of Moebius syndrome remains unknown. It is interesting that the clinical manifestations of Moebius syndrome can include CBVCP. However, the pathophysiology of CBVCP is unknown and further investigations into the etiology of Moebius syndrome are required. PMID- 17136553 TI - Traplining in bumblebees (Bombus impatiens): a foraging strategy's ontogeny and the importance of spatial reference memory in short-range foraging. AB - To test the relative importance of long-term and working spatial memories in short-range foraging in bumblebees, we compared the performance of two groups of bees. One group foraged in a stable array of six flowers for 40 foraging bouts, thereby enabling it to establish a long-term memory of the array, and adjust its spatial movements accordingly. The other group was faced with an array that changed between (but not within) foraging bouts, and thus had only access to a working memory of the flowers that had been visited. Bees in the stable array started out sampling a variety of routes, but their tendency to visit flowers in a repeatable, stable order ("traplining") increased drastically with experience. These bees used shorter routes and converged on four popular paths. However, these routes were mainly formed through linking pairs of flowers by near neighbour movements, rather than attempting to minimize overall travel distance. Individuals had variations to a primary sequence, where some bees used a major sequence most often, followed by a minor less used route, and others used two different routes with equal frequency. Even though bees foraging in the spatially randomized array had access to both spatial working memory and scent marks, this manipulation greatly disrupted foraging efficiency, mainly via an increase in revisitation to previously emptied flowers and substantially longer search times. Hence, a stable reference frame greatly improves foraging even for bees in relatively small arrays of flowers. PMID- 17136554 TI - [Airway leak pressure of the ProSeal laryngeal mask airway. Comparison with and without a gastric tube placed through the drain-tube]. AB - BACKGROUND: The ProSeal laryngeal mask airway (PLMA) has been studied in numerous investigations and the airway leak pressure (P(leak)) is often used as a primary end-point, particularly in comparative studies with other supraglottic airway devices. The PLMA offers the opportunity to place a gastric tube through the drain-tube and P(leak) measurement can take place both with and without a gastric tube. With this study we tested the hypothesis that the use of a gastric tube influences the P(leak). METHODS: The P(leak) of the PLMA was studied in 98 patients under total intravenous anaesthesia with propofol (0.1-0.15 mg*kgBW 1*min-1) and remifentanil (0.1-0.3 microg*kgBW-1*min-1) before and after placement of a gastric tube through the drain-tube of the PLMA. RESULTS: There was no significant difference between the mean baseline P(leak) without a gastric tube in place (25+/-6.3 cm H(2)O) and the mean P(leak) after placement of a gastric tube (25+/-6.7 cm H2O; p=0.6). CONCLUSION: Placement of a gastric tube through the drain-tube of the PLMA does not influence the P(leak). Providing the same method of measurement has been used, P(leak) values from different studies obtained with or without a gastric tube in place are comparable. PMID- 17136555 TI - [Tonsillectomy technique: bipolar scissors vs raspatory: results of a case control study in 138 patients]. AB - BACKGROUND: Postoperative bleeding is the major complication in tonsillectomy, and pain the most common side effect. The use of bipolar scissors versus blunt dissection tonsillectomy were compared in this study in order to evaluate postoperative bleeding and pain, as well as operative time. METHODS: In this case control study, 138 patients with the diagnosis of chronic tonsillitis, mononucleosis or a peritonsillar abscess were divided into two groups. A total of 78 patients were operated using bipolar scissors while 60 patients underwent tonsillectomy by blunt dissection. Operating time, frequency of postoperative bleeding and the postoperative pain score were compared between these two groups. RESULTS: The average operating time in the bipolar scissor group showed a tendency to be shorter than in the blunt dissection group (mean 4.1 min), although this did not reach a level of statistical significance. No differences were seen in pain scores or in the incidence of postoperative bleeding. CONCLUSION: The data documented in this study show that tonsillectomy with bipolar scissors might represent a surgical option to reduce surgical time in a larger patient group. Postoperative pain and the incidence of postoperative bleeding did not show any statistical difference between the two surgical techniques. PMID- 17136556 TI - [Escalating immunomodulatory therapy of multiple sclerosis. Update (September 2006)]. AB - The updated recommendations presented here reflect new developments in the diagnostic work-up and immunotherapy of multiple sclerosis (MS) as well as optimization of medical care for MS patients. Monoclonal antibodies provide considerable improvement of treatment, but their use in basic therapy is restricted by their side effect profile. Thus, for the time being, natalizumab is only approved for monotherapy after basic treatment has failed or for rapidly progressive relapsing-remitting MS. In contrast, long-term data on recombinant beta-interferons and glatiramer acetate (Copaxone) show that even after several years no unexpected side effects occur and that a prolonged therapeutic effect can be assumed which correlates with the dose or frequency of treatment. Recently IFN-beta1b (Betaferon) was approved for prophylactic treatment after the first attack (clinically isolated syndrome, CIS). During treatment with beta interferons, neutralizing antibodies can emerge with possible loss of effectivity. In contrast, antibodies play no role in treatment with glatiramer acetate. During or after therapy with mitoxantrone, serious side effects (cardiomyopathy, acute myeloid leukemia) appeared in 0.2-0.4% of cases. Plasmapheresis is limited to individual curative attempts in escalating therapy of a severe attack. According to the revised McDonald criteria, the diagnosis of MS can be made as early as the occurrence of the first attack (CIS). Recommendations for optimized care of MS patients are also new, thus implementing a resolution of the European Parliament. PMID- 17136559 TI - Sonography of the superomedial part of the spring ligament complex of the foot: a study of cadavers and asymptomatic volunteers. AB - OBJECTIVE: To evaluate the ability of high-resolution sonography for assessing the thickness and echogenicity of the superomedial part of the normal spring ligament in the foot in cadavers and asymptomatic volunteers. MATERIALS AND METHODS: The superomedial part of the normal spring ligament of four cadaveric feet was imaged with a high-resolution linear array transducer. Upon localization, the ligament was injected with 0.1% methylene blue with sonographic guidance. A posteromedial approach was used to dissect the feet immediately following injection to confirm accurate identification of the ligament. The bilateral ligaments in 40 asymptomatic adult volunteers were subsequently imaged. RESULTS: Surgical dissection confirmed the accurate injection of methylene blue into all four cadaveric ligaments. The superomedial part of the normal spring ligament was identified bilaterally in all of the 40 asymptomatic volunteers, with a mean thickness of 3 mm in longitudinal short axes. The ligament was echogenic relative to surrounding fat, and it was thinner in women. Differences in ligament measurements with respect to age, sex, side (left vs. right) and foot dominance were not significant. The volunteers' height, weight and body mass index had a weakly positive correlation with ligament thickness. CONCLUSION: High resolution ultrasound can be used to identify and measure the thickness of the superomedial part of the normal spring ligament. The provided baseline measurements for the normal ligament could prove valuable when assessing the abnormal ligament. PMID- 17136560 TI - Comparison of CT and MRI in patients with tibial plateau fracture: can CT findings predict ligament tear or meniscal injury? AB - OBJECTIVE: (1) To determine the accuracy of computed tomography (CT) in the evaluation of ligament tear and avulsion in patients with tibial plateau fracture. (2) To evaluate whether the presence or severity of fracture gap and articular depression can predict meniscal injury. DESIGN AND PATIENTS: A fellowship-trained musculoskeletal radiologist retrospectively reviewed knee CT and MRI examinations of 41 consecutive patients presenting to a level 1 trauma center with tibial plateau fractures. Fracture gap, articular depression, ligament tear and footprint avulsions were assessed on CT examinations. The MRI studies were examined for osseous and soft tissue injuries, including meniscal tear, meniscal displacement, ligament tear, and ligament avulsion. RESULTS: CT demonstrated torn ligaments with 80% sensitivity and 98% specificity. Only 2% of ligaments deemed intact on careful CT evaluation had partial or complete tears on MRI. Although the degree of fracture gap and articular depression was significantly greater in patients with meniscal injury compared with those without meniscal injury, ROC analysis demonstrated no clear threshold for gap or depression that yielded a combination of high sensitivity and specificity. CONCLUSIONS: In the acute setting, CT offers high sensitivity and specificity for depicting osseous avulsions, as well as high negative predictive value for excluding ligament injury. However, MRI remains necessary for the preoperative detection of meniscal injury. PMID- 17136562 TI - Systemic lupus erythematosus: a genetic epidemiology study of 695 patients from China. AB - Our purpose was to explore potential genetic models for systemic lupus erythematosus (SLE) and analyze genetic epidemiologic characteristics of SLE in a Chinese population. Data for 695 patients with SLE were obtained by using a uniform questionnaire. Patients, clinical characteristics and their family history were analyzed using software. A complex segregation analysis was conducted to propose potential genetic models for SLE. The mean +/- SD age of onset were 30.2 +/- 10.5 years and mean time to progression to SLE was 32.5 +/- 44.4 months. The most frequent initial manifestations were malar rash (61.3%). During the evolution of the disease, the main clinical features were arthritis in 73.6% of our patients, followed by malar rash (68.1%), and renal involvement (56.7%). As the first symptom, the late-onset group (onset of disease beyond the age of 50 years) less often showed malar rash (45% vs. 63.4% in the early-onset group; p = 0.001). There were no significant differences in the other cumulative clinical symptoms between late-onset and early-onset group, except for a lower prevalence of malar rash, photosensitivity and alopecia and a higher prevalence of mucosal ulcers in the late-onset group. A positive family history of SLE was obtained in 50 patients (7.2%). There were no statistical differences in clinical characteristics between familial SLE and sporadic SLE patients. The heritability of SLE was 43.6%, the genetic model of SLE could be polygenetic model and major gene mode is the best fitted one. SLE could be a multifactorial disease with polygenetic model. PMID- 17136563 TI - Explorative immunohistochemical study to evaluate the addition of a topical corticosteroid in the early phase of alefacept treatment for psoriasis. AB - The aim of this study was to explore the additional effect of betamethasone dipropionate cream in the early phase of an intramuscular (IM) alefacept course, on plaque severity and on modulating T-cell subsets, cells expressing NK receptors, epidermal proliferation and keratinocyte differentiation in lesional psoriatic skin. Therefore, sixteen patients with moderate-to-severe chronic plaque psoriasis received 15 mg alefacept IM for 12 weeks, followed by a 12-week follow-up period. The first 4 weeks, patients were randomized 1:1 to either betamethasone dipropionate, or the vehicle cream, once daily. Plaque severity (SUM) was assessed and serial biopsies were immunohistochemically stained for T cell subsets (CD3, CD4, CD8, CD45RO, CD45RA, CD2, CD25, GITR), cells expressing NK-receptors (CD94 and CD161), epidermal proliferation (Ki67) and differentiation (K10), which were quantified using manual and digital image analysis. Alefacept monotherapy resulted in statistically significant improvement in plaque severity. Subsequently, immunohistochemical assessments on T-cell subsets, epidermal proliferation (Ki67) and keratinization (K10) revealed marked time-related improvements with respect to the mentioned parameters, without significant differences between both treatment regimens. Alefacept monotherapy induces improvement of plaque severity, which is accompanied by a reduction in activated (CD2+, CD25+, CD45RO+) dermal CD4+ and activated epidermal CD8+ T cells, epidermal proliferation and differentiation. Once daily treatment with betamethasone dipropionate cream during the first 4 weeks of an intramuscular alefacept course did not provide substantial additional clinical and immunohistochemical improvement. PMID- 17136564 TI - Innovative restriction site created PCR-RFLP for detection of benzimidazole resistance in Teladorsagia circumcincta. AB - Benzimidazole compounds, especially albendazole, are the most commonly used anthelmintics for deworming of small ruminants in Iran. It is believed that the therapeutic effects of the benzimidazoles (BZs) come through their binding capacity to the beta-tubulin isotype 1. Substitution of phenylalanine to tyrosine at position 200 of this polypeptide confers resistance to BZs. Several investigators developed different biological- and molecular-based techniques to demonstrate the occurrence of resistance in helminthes against BZs. To address the determination of resistance at position 200 of beta-tubulin isotype 1, we developed an innovative restriction site created polymerase chain reaction restriction fragment length polymorphism, in which nucleotide A at the position of 637 upstream flanked by the first two coding sequences of the phenylalanine (TT) triplet was substituted through the nucleotide G. The introduced modification in forward primer (UTvet MF-primer) leads to the creation of restriction site (AACGTT) for PSP1. Therefore, in the case of normal allele only, PSP1 can cut the corresponding PCR product. In the first step, the genomic DNA was isolated from each single Teladorsagia circumcincta collected either from the abomasa of untreated (n = 35) or of 5 mg/kg BW 2.5% albendazole suspension treated (n = 40) sheep. It was amplified with the primer pair, creating PCR product of 403 bp in length. In the second step, the PCR product was extracted from agarose gel and amplified with the modified forward primer (UTvet MF-primer) and the same reverse primer as in step 1, creating a PCR product of 222 bp. The PCR product was then cut with PSP1 to obtain in the case of normal allele two DNA products (183 and 39 bp). Eight of the 35 worms collected from the untreated sheep were BZ(SS) homozygotes, and the rest (27) were BZ(RS) heterozygotes. In our preliminary experiment, we could not find a BZ(RR) homozygote form within the examined samples. Five out of 40 worms collected from the albendazole-treated sheep were BZ(RR) homozygotes, whereas the rest (35) were BZ(RS) heterozygotes. No BZ(SS) homozygote form was detected within this group. PMID- 17136565 TI - The postnatal development of the optic nerve of a reptile (Vipera aspis): A quantitative ultrastructural study. AB - The number of axons in the optic nerve of the ovoviviparous reptile Vipera aspis was estimated from electron micrographs taken during the first 5 weeks of postnatal life. One to two days after birth, the optic nerve contains about 170,000 fibres, of which about 9% are myelinated. At the end of the fifth postnatal week, the number of optic fibres has fallen to about 100,000, of which about 42% are myelinated. This fibre loss continues after the fifth postnatal week, since in the adult viper the nerve contains about 60,000 fibres, of which 85% are myelinated; overall, about 65% of the optic nerve fibres present at birth disappear before the number of axons stabilises at the adult level. This study shows, for the first time, that the mode of development of the visual axons of reptiles is not that of anamniote vertebrates but similar to that of birds and mammals. PMID- 17136567 TI - Cytoskeletal structures, ultrastructural characteristics and the capsule of the basidiomycetous yeast Cryptococcus laurentii. AB - The cytoskeleton, capsule and cell ultrastructure were studied during the cell cycle of Cryptococcus laurentii. In an encapsulated strain, cytoplasmic microtubules and a mitotic spindle were detected. Mitosis was preceded by migration of the nucleus into the bud. F-actin failed to be visualised by rhodamine-phalloidin (RhPh) in encapsulated cells and therefore an acapsular strain was used. The following actin structures were found: actin dots, actin cables and cytokinetic ring. Ultrastructural studies showed the presence of a nucleus in the bud before mitosis. A collar-shaped structure was seen at the base of bud emergence. A lamellar cell wall and a rough outer surface of the cells were detected. Cytoskeletal structures found in C. laurentii are similar to those in Cryptococcus neoformans, which is a serious human pathogen. PMID- 17136568 TI - Development of a recombinant strain of Bacillus thuringiensis subsp. kurstaki HD 73 that produces the endochitinase ChiA74. AB - Bacillus thuringiensis subsp. kurstaki HD-73 was transformed with the homologous endochitinase gene chiA74 of B. thuringiensis subsp. kenyae LBIT-82 under the regulation of its own promoter and Shine-Dalgarno sequence. The plasmid, pEHchiA74, which harbors chiA74, was detected by southern blot analysis and showed high segregational stability when the recombinant strain was grown in a medium without antibiotic. The recombinant bacterium transformed with pEHchiA74 showed an improvement in chitinolytic activity three times that of the wild-type strain. Expression of ChiA74 did not have any deleterious effect on the crystal morphology and size, but sporulation and Cry1Ac production in rich medium (nutrient broth with glucose) was reduced by approximately 30%. No significant increase in the toxicity of the transformant bacterium toward Plutella xylostella was detected using the same amount of total protein. However, it is possible that ChiA74 synthesis compensated for the decrease in net Cry1Ac synthesis and toxicity observed with the recombinant strain. PMID- 17136569 TI - Enhancement of growth and polysaccharide production in suspension cultures of protocorm-like bodies from Dendrobium huoshanense by the addition of putrescine. AB - Putrescine at 0.6 mM stimulated protocorm-like body growth and polysaccharide synthesis in suspension cultures of Dendrobium huoshanense. The specific growth rate of protocorm-like body increased from 0.047 to 0.056 day(-1), and the maximum dry weight and polysaccharide production reached 33.2 and 2.94 g l(-1), respectively, while they were 24.6 and 2.12 g l(-1), respectively, in the control. The administration of polyamine inhibitor, alpha-DL difluoromethylarginine, at 1 mM, decreased protocorm-like body growth and polysaccharide production to 21.4 and 1.76 g l(-1), respectively. PMID- 17136570 TI - TNT transformation products are affected by the growth conditions of Raoultella terrigena. AB - High concentrations of 2,4,6-trinitrotoluene (TNT) and related nitroaromatic compounds are commonly found in soil and groundwater at former explosive plants. The bacterium, Raoultella terrigena strain HB, isolated from a contaminated site, converts TNT into the corresponding amino products. Radio-HPLC analysis with [(14)C]TNT identified aminodinitrotoluene, diaminonitrotoluene and azoxy-dimers as the main metabolites. Transformation rate and the type of metabolites that predominated in the culture medium and within the cells were significantly influenced by the culture conditions. The NAD(P)H-dependent enzymatic reduction of nitro-substituted compounds by cell-free extracts of R. terrigena was evaluated in vitro. PMID- 17136571 TI - Isolation and characterization of a novel mycovirus, PeSV, in Pleurotus eryngii and the development of a diagnostic system for it. AB - A novel mycovirus was isolated from a cultivated edible mushroom, Pleurotus eryngii, with severe epidemic symptoms. Purification of the virus was carried out by a sequential procedure of polyethylene glycol precipitation, differential centrifugation, and equilibrium centrifugation in a CsCl gradient. Nuclease digestion assay and protein analysis revealed that the virus consisted of a single-stranded RNA (ssRNA) genome of 7.8 kbp which was encapsulated by a coat protein of 22 kDa. Transmission electron microscope showed that it was spherical with a diameter of 31 nm. Since there was neither a previous report on discovery of a virus in P. eryngii, nor known mushroom viruses with similar characteristics, we concluded that this is a novel virus and thus have named it as P. e ryngii Spherical Virus (PeSV). Because of a diagnostic test would be helpful in preventing the PeSV-related disease outbreaks, we developed a triple antibody sandwich-ELISA (TAS-ELISA) system using anti-PeSV mouse monoclonal and anti-PeSV rabbit polyclonal antibodies. The TAS-ELISA system successfully detected less than 0.5 microg of the virus particles in 1 g diseased mushroom tissue collected from various commercial farms. PMID- 17136572 TI - Efficient purification of transglutaminase from recombinant Streptomyces platensis at various scales. AB - An efficient system for the fast and efficient purification of transglutaminase from recombinant Streptomyces platensis and expressed in Streptomyces lividans 25 2 is described. Because the purification procedure of this system is flexible, culture broth from laboratory (20 l) and pilot-plant (130 l) fermentations were used to purify the enzyme to electrophoretic homogeneity with high purity (90 95%) and yield (61-77%) within 1 or 2 days. PMID- 17136573 TI - Use of DNA marker to select well-adapted Phaseolus-symbionts strains under acid conditions and high temperature. AB - Soil acidity and high temperature contribute to the failure of nodulation in the common bean. It is therefore urgent to select strains with a high competitive ability under these stress conditions. Two Egyptian Rhizobium etli strains, EBRI 2 and EBRI 26, were examined against Rhizobium tropici CIAT 899G labeled with the gus (beta-glucuronidase) reporter gene. EBRI 2 and EBRI 26 were less competitive than CIAT 899G under acid conditions with both the Egyptian cultivar Giza 3 and the Colombian cultivar Rab 39. However, EBRI 2 and EBRI 26 gave higher nodule occupancy (78% and 62.5, respectively) than the nodule occupancy (18.5% and 35%) obtained by CIAT 899G at 35 degrees C with cultivar Giza 3. Soil acidity (pH 5.8) was less detrimental to the nodule occupancy of EBRI 2 than EBRI 26 when they tested in competition with CIAT 899G. PMID- 17136574 TI - The application of a murine bone bioreactor as a model of tumor: bone interaction. AB - A limited number of in vivo models that rapidly assess bone development or allow for the study of tumor progression in a closed in vivo environment exist. To address this, we have used bone tissue engineering techniques to generate a murine in vivo bone bioreactor. The bioreactor was created by implanting an osteoconductive hydroxyapatite scaffold pre-loaded with saline as a control or with bone morphogenetic protein-2 (BMP-2) to the murine femoral artery. Control and BMP-2 bioreactors were harvested and histologically assessed for vascularization and bone formation at 6 and 12 weeks post implantation. BMP-2 significantly enhanced the formation of osteoid within the bioreactor in comparison to the controls. To test the in vivo bone bioreactor as a model of tumor: bone interaction, FVB mice were implanted with control or BMP-2 treated bioreactors. After 6 weeks, an osteolytic inducing mammary tumor cell line tagged with luciferase (PyMT-Luc) derived from the polyoma virus middle T (PyMT) model of mammary tumorigenesis was delivered to the bioreactor via the femoral artery. Analysis of luciferase expression over time demonstrated that the presence of osteoid in the BMP-2 treated bioreactors significantly enhanced the growth rate of the PyMT-Luc cells in comparison to the control group. These data present a unique in vivo model of ectopic bone formation that can be manipulated to address molecular questions that pertain to bone development and tumor progression in a bone environment. PMID- 17136575 TI - Host matrix metalloproteinase-9 contributes to tumor vascularization without affecting tumor growth in a model of prostate cancer bone metastasis. AB - Matrix metalloproteinases (MMPs) have been associated with initiation, progression and vascularization of a number of tumors. However, clinical trials using MMP inhibitors failed to meet expectations. Previously, we demonstrated the potential importance of MMP-9 activity in experimental prostate cancer bone tumor tissue. However, the particular roles of host- and tumor-derived MMP-9 remains to be defined. Herein, we examined the role of host MMP-9 in subcutaneous and intraosseous growth of the human androgen independent prostate cancer cell line PC3 in MMP-9 deficient mice. In the subcutaneous model, the tumor incidence in the control (RAG-1(ko/ko)) and experimental (RAG-1(ko/ko) /MMP-9(ko/ko)) group was 100%, with similar tumor growth kinetics and microvascular densities. In the intraosseous tumor model, the tumor incidence was higher in RAG-1(ko/ko) /MMP 9(ko/ko mice than in RAG-1(ko/ko) mice (67% and 39%, respectively), though no statistical differences were found. The intraosseous tumor areas were similar in both groups, and the number of tumor-associated osteoclasts did not differ significantly. However, the microvascular density of intraosseous tumors was higher in RAG-1(ko/ko) than in RAG-1(ko/ko)/MMP-9(ko/ko) mice, though no changes in tumor growth could be detected. In an in vitro assay, we found that bone marrow (BM) cells increased the invasiveness of PC3 cells, and that this enhancement was independent of MMP-9 expression by marrow cells. Our results with the RAG-1 model suggest that host-derived MMP-9 is neither necessary nor sufficient for subcutaneous or intraosseous PC3 tumor growth, osteoclastic response, or in vitro invasiveness of tumor cells. PMID- 17136576 TI - Visualization of nascent tumor angiogenesis in lung and liver metastasis by differential dual-color fluorescence imaging in nestin-linked-GFP mice. AB - Nestin regulatory-element-driven green fluorescent protein (ND-GFP) transgenic mice highly express GFP in proliferating endothelial cells and nascent blood vessels. In the present study, we visualized angiogenesis in experimental lung and liver metastases by GFP imaging in the ND-GFP transgenic mice. The murine melanoma cell line, B16F10 expressing red fluorescent protein (RFP), was injected i.v. in ND-GFP mice. ND-GFP was highly expressed in proliferating nascent blood vessels in the tumors that developed in the lung after tail vein injection, and in the tumors that developed in the liver after portal vein injection of RFP expressing melanoma cells. Liver metastasis and angiogenesis were imaged intravitally. Doxorubicin significantly decreased metastatic angiogenesis in the liver. These results demonstrate a new imageable model of angiogenesis in metastasis in the liver and the lung. This new model should enable further understanding of the onset of angiogenesis in metastasis and its effect on metastatic growth. The model will serve as a unique screen for inhibitors of angiogenesis of metastatic tumors. The fact that liver-metastasis angiogenesis can be imaged in the live animal enables real-time studies of the effect of angiogenesis inhibitors. PMID- 17136577 TI - Inversion polymorphism and a new polytene chromosome map of Zaprionus indianus Gupta (1970) (Diptera: Drosophilidae). AB - Zaprionus indianus is a recent invader in Brazil and was probably introduced from the West Afrotropical zone. So far, studies regarding its chromosomal polymorphism were limited to India. We found that Brazilian populations were very different from Indian ones. Five new inversions have been discovered. In(II)A, already described in India, where it is quite common, has also been found in Brazil, where it is very rare. The X-chromosome has three inversions; In(X)Na, In(X)Ke and In(X)Eg, which are frequent in all Brazilian populations studied. In every case, we observed strong linkage disequilibrium among these gene arrangements. During the primary collection period (2001-2002), we noticed a significant positive correlation between the frequency of these inversions and latitude, but this was not confirmed in later investigations. Rearrangement In(IV)EF was also common in all populations, while inversion In(V)B was only found in southern populations. Our data suggest that the founders that recently invaded Brazil were polymorphic for the six inversions observed. The place of origin might be identified more precisely by investigating West African populations. In order to facilitate further investigations, we present an updated polytene chromosome photomap, locating the breakpoints of every inversion observed in Brazilian populations. PMID- 17136578 TI - Karyotypic characterization and genomic organization of the 5S rDNA in Erpetoichthys calabaricus (Osteichthyes, Polypteridae). AB - Polypterids are a group of Osteichthyan fish whose evolutionary relationships with closer basal ray-finned and lobe-finned fish have been disputed since their discovery. Very little is known about the evolutive karyology in the whole Polypteriformes group. In order to fill this gap, a cytogenetic analysis of Erpetoichthys calabaricus species was performed, using both classical and molecular techniques. Karyotype structure (2n = 36; FN = 72), chromosome location of telomeric sequences (TTAGGG)n and ribosomal 5S and 18S rRNA genes were examined in twenty specimens of E. calabaricus by using Ag-NOR, classical C banding, sequential CMA3/4',6-diaminidino-2-phenylindole (DAPI) staining and fluorescent in situ hybridization (FISH). CMA3 marked all centromerical and some (no. 1 and no. 15) telomeric regions. Staining with Ag-NOR and CMA3 showed the presence of two NORs on the p arm of the chromosome pair no. 1. Hybridization with telomeric probes (TTAGGG)n showed signals at the end of all chromosomes. 5S rDNA was cloned and sequenced. After the alignment, the 5S rRNA sequences revealed an organization made up of two different classes of tandem arrays (type I and type II). FISH with 5S rDNA marked the telomeric regions of the small chromosome pair no. 15, while FISH with 18S rDNA marked the telomeric region of the pair no. 1. The results obtained were compared with cariological data on closer species now available in literature. PMID- 17136579 TI - Genes in the postgenomic era. AB - We outline three very different concepts of the gene-instrumental, nominal, and postgenomic. The instrumental gene has a critical role in the construction and interpretation of experiments in which the relationship between genotype and phenotype is explored via hybridization between organisms or directly between nucleic acid molecules. It also plays an important theoretical role in the foundations of disciplines such as quantitative genetics and population genetics. The nominal gene is a critical practical tool, allowing stable communication between bioscientists in a wide range of fields grounded in well-defined sequences of nucleotides, but this concept does not embody major theoretical insights into genome structure or function. The post-genomic gene embodies the continuing project of understanding how genome structure supports genome function, but with a deflationary picture of the gene as a structural unit. This final concept of the gene poses a significant challenge to conventional assumptions about the relationship between genome structure and function, and between genotype and phenotype. PMID- 17136580 TI - The influence of matrix attachment regions on transgene expression in Arabidopsis thaliana wild type and gene silencing mutants. AB - Many studies in both animal and plant systems have shown that matrix attachment regions (MARs) can increase the expression of flanking transgenes. However, our previous studies revealed no effect of the chicken lysozyme MARs (chiMARs) on transgene expression in the first generation transgenic Arabidopsis thaliana plants transformed with a beta-glucuronidase gene (uidA) unless gene silencing mutants were used as genetic background for transformation. In the present study, we investigated why chiMARs do not influence transgene expression in transgenic wild-type Arabidopsis plants. We first studied the effect of chiMARs on transgene expression in the progeny of primary transformants harboring chiMAR-flanked T DNAs. Our data indicate that chiMARs do not affect transgene expression in consecutive generations of wild-type A. thaliana plants. Next, we examined whether these observed results in A. thaliana transformants are influenced by the applied transformation method. The results from in vitro transformed A. thaliana plants are in accordance with those from in planta transformed A. thaliana plants and again reveal no influence of chiMARs on transgene expression in A. thaliana wild-type transformants. The effect of chi-MARs on transgene expression is also examined in in vitro transformed Nicotiana tabacum plants, but as for A. thaliana, the transgene expression in tobacco transformants is not altered by the presence of chi-MARs. Taken together, our results show that the applied method or the plant species used for transformation does not influence whether and how chiMARs have an effect on transgene expression. Finally, we studied the effect of MARs (tabMARs) of plant origin (tobacco) on the transgene expression in A. thaliana wild-type plants and suppressed gene silencing (sgs2) mutants. Our results clearly show that similar to chiMARs, the tobacco-derived MARs do not enhance transgene expression in a wild-type background but can be used to enhance transgene expression in a mutant impaired in gene silencing. PMID- 17136581 TI - An unusual case of ureteral tumor in a duplex system. AB - We report an interesting case of ureteral tumor involving both limbs of an incompletely duplicated ureter. Such a case has not been reported in the literature. A 51-year-old female presented with refractory hematuria. A complete evaluation revealed an incompletely duplicated right system with an isolated distal ureteral tumor extending proximally into both arms of the ureter. The patient underwent a successful right nephroureterectomy. Pathology revealed low grade papillary urothelial cancer confined to both arms of the ureter. A brief review of the literature and management is detailed herein. PMID- 17136582 TI - Dilemmas in the management of female stress incontinence: the role of pelvic floor muscle training. AB - Treatment options for female stress urinary incontinence include pelvic floor muscle training, lifestyle interventions, bladder retraining, pharmacotherapy, anti-incontinence devices and surgery. Several consensus statements recommend pelvic floor muscle training as first line treatment. The aim of this review is to analyse all the currently available data and propose a treatment algorithm for clinical practice. A literature-based critical presentation of all treatment modalities, methods of assessing efficacy and comparison between them using a patient-centered approach was made. Many of the studies are observational, non randomized with several methodological problems that lead to confusion. Emphasis was made to high quality randomized trials. The proposed treatment algorithm established only on evidence-based data. Management strategy however, must identify patient expectations and involve them in the decision-making more than traditional measures of treatment success. PMID- 17136583 TI - Autonomic side effects of Botulinum toxin Type B intravesical injections: report of 4 cases and review of the literature. AB - Recently, intravesical injections of Botulinum toxin type B (BT-B) have been trialled in an attempt to treat detrusor overactivity. Use of Botulinum toxin A (BT.-A), for the same in the past, has been quite successful without many reported side effects. Soon after commencing the clinical use of BT-B, a side effect profile was noticed not reported with BT-A. A total of 20 patients with refractory detrusor overactivity were included in this randomized, double blind, placebo controlled, cross over trial. As per protocol, 5000 MU (1 ml) BT-B diluted to a volume of 20 mls with normal saline were injected in 10 different sites into the detrusor, sparing the trigone. We report four cases of autonomic side effects after these injections which strongly suggest clinically relevant systemic spread of the toxin. PMID- 17136584 TI - Spontaneous bladder rupture caused by a giant vesical calculus. AB - Spontaneous rupture of the urinary bladder is an uncommon occurrence. A 36-year old man had complaints of pain and progressive distension of abdomen and anuria for 2 days. His abdomen was tense, tender and distended with free fluid. Blood urea was 340 mg% and ascitic fluid urea was 337 mg%. An USG showed massive ascitis, a large vesical calculus and a left renal calculus. The urinary bladder could not be catheterized. Patient underwent hemodialysis and placement of abdominal drains. About 2 l of yellow turbid fluid was drained. Cystolithotomy showed a 6 cm size impacted calculus with a rent in the dome of the bladder, which was repaired. Subsequently patient underwent percutaneous nephrolithotrypsy for left staghorn renal calculus and nephrectomy for right non-functioning kidney. PMID- 17136586 TI - Frequency, nature and determinants of pharmaceutical consultations provided in private by Dutch community pharmacists. AB - OBJECTIVE: According to a report published by the federation of Dutch patients' associations, patients would like to see a pharmacist, who acts more as a personal adviser. This raised the question, how often Dutch community pharmacists have personal consultations with their patients in daily practice, on which factors this depends, and what kind of topics are discussed during these meetings. SETTING: Community pharmacies in the Netherlands. METHOD: A questionnaire was distributed among 800 randomly selected pharmacies. Questions were restricted to consultations characterized by one-to-one contact, drug therapy related content, and adequate privacy. These consultations were labelled as pharmaceutical consultations in private to distinguish them from other contacts between pharmacists and patients. MAIN OUTCOME MEASURE: Number, content, and character of consultations. RESULTS: 198 (24.8%) community pharmacies responded. The pharmacists provide an average of roughly 1.2 consultations in private per working day. The vast majority of respondents provided face-to-face and telephone consultations (94.4 and 91.9%, respectively), only a minority gave consultations by e-mail (30.8%). These consultations primarily dealt with topics related to medication safety. The mean overall time spent was 290 min per month. A relatively high frequency of personal consultations was significantly associated with the absolute number of full-time equivalent pharmacists in the pharmacy. CONCLUSION: The frequency of pharmaceutical consultations in private is low, but may be improved by reorganisation of the pharmacist's activities. The possibility of personal consultations by e-mail is not yet well-developed. Further research is needed to assess the patient's view of pharmaceutical consultations in private. PMID- 17136587 TI - Human physique and sexual attractiveness: sexual preferences of men and women in Bakossiland, Cameroon. AB - Men and women living in a rural community in Bakossiland, Cameroon were asked to rate the attractiveness of images of male or female figures manipulated to vary in somatotype, waist-to-hip ratio (WHR), secondary sexual traits, and other features. In Study 1, women rated mesomorphic (muscular) and average male somatotypes as most attractive, followed by ectomorphic (slim) and endomorphic (heavily built) figures. In Study 2, amount and distribution of masculine trunk (chest and abdominal) hair was altered progressively in a series of front-posed male figures. A significant preference for one of these images was found, but the most hirsute figure was not judged as most attractive. Study 3 assessed attractiveness of front-posed male figures which varied only in length of the non erect penis. Extremes of penile size (smallest and largest of five images) were rated as significantly less attractive than three intermediate sizes. In Study 4, Bakossi men rated the attractiveness of back-posed female images varying in WHR (from 0.5-1.0). The 0.8 WHR figure was rated markedly more attractive than others. Study 5 rated the attractiveness of female skin color. Men expressed no consistent preference for either lighter or darker female figures. These results are the first of their kind reported for a Central African community and provide a useful cross-cultural perspective to published accounts on sexual selection, human morphology and attractiveness in the U.S., Europe, and elsewhere. PMID- 17136588 TI - Effects of sexual arousal on genital and non-genital sensation: a comparison of women with vulvar vestibulitis syndrome and healthy controls. AB - The relationship between sexual arousal and sensory perception has been a topic largely neglected within the realm of human sexuality research. The present study assessed the influence of sexual arousal on genital and non-genital sensation in women. It also examined the theory that painful intercourse is associated with insufficient sexual arousal. A total of 20 healthy women and 20 women with Vulvar Vestibulitis Syndrome (VVS) underwent genital and non-genital sensory testing at baseline and in response to erotic and neutral stimulus films. Touch and pain thresholds were assessed at the vulvar vestibule, inside the labia minora, and on the volar surface of the forearm. Sexual arousal was assessed via the measurement of surface skin temperature changes of the labia minora using a labial thermistor clip. Participants also completed questionnaires pertaining to mood, pain, and sexual functioning. In response to the erotic stimulus, both groups evidenced a significant increase in physiological sexual arousal and vulvar sensitivity. Women with VVS reported a significantly lower desire to engage in intercourse after having viewed the erotic film and reported lower levels of desire and arousal on questionnaire measures. Women with VVS also exhibited significantly more genital and non-genital pain sensitivity than healthy women across all conditions, in addition to more catastrophizing, hypervigilance, and fear of pain. Contrary to some theories, these data suggest that women with VVS are not lacking in physiological sexual arousal, and that physiological sexual arousal may actually increase vulvar sensation. Lack of subjective sexual arousal, however, may yet be implicated in vulvar pain during intercourse. PMID- 17136589 TI - Body piercing, personality, and sexual behavior. AB - The associations of body piercing with other social characteristics, personality, and sexual behavior were investigated in a population-based sample of young adults, in light of the theory that body piercing has meaning in terms of a corporeal expression of the self. At age 26 years, 966 (95%) of 1019 members of the birth-cohort of the Dunedin Multidisciplinary Health and Development Study were asked about body piercing (at interview) and sexual behavior (questions presented by computer). Assessment of personality traits was conducted at ages 18 or 21 years. In total, 183 participants (9% of the men and 29% of the women) had piercings at a site other than the earlobes. People who lived outside New Zealand or who were of Maori descent were more likely to be pierced, but unemployment and low occupational status were not significantly related to piercing. Women who were pierced, compared with those without piercings, were more likely to have personality traits of low constraint or high negative emotionality. Women with piercings were also more likely to report having had, during the previous year, five or more heterosexual partners (odds ratio, 5.8, 95% CI: 2.3-14.6) or any same-sex partner involving genital contact (odds ratio, 10.3, CI: 2.9-37.2). The associations with sexual behavior in men were weaker and not statistically significant. In this population, body piercing in women was associated with sexual behavior. Having multiple heterosexual partners or any same-sex partner was very rare among women without piercings. The theory of meaning for body piercing was generally supported, offering the possibility of a richer understanding of this phenomenon in the general population. PMID- 17136590 TI - Sexual desire and linguistic analysis: a comparison of sexually-abused and non abused women. AB - Although studies have identified a relationship between a history of child sexual abuse (CSA) and problems with hypoactive sexual desire, little is known about the potential cognitive and affective mechanisms involved in the sexual desire of women with a history of CSA. In this study, 27 women with a history of CSA and 22 women with no history of abuse were asked to write about sexual and non sexual topics. The Linguistic Inquiry Word Count software program was used to compute the percentage of words that fell into positive emotions, negative emotions, body, and sex categories. As expected, women with a history of CSA used more negative emotions words when writing about sexual topics, but not non-sexual topics, compared to non-abused women. Women with a history of CSA also used more sex words when writing about the non-sexual topics compared to non-abused women. Frequencies of body and sex words used in the sexual texts were positively linked to levels of sexual desire function. This association was not different between women with and without a history of CSA. A history of CSA remained an independent predictor of levels of sexual desire dysfunction even when taking into consideration the language used in the sexual texts, indicating that there may be aspects of the sexual desire experienced by women with a history of CSA that differ from non-abused women that remain unexplored. PMID- 17136591 TI - Subjective and objective measures of attractiveness and their relation to sexual behavior and sexual attitudes in university students. AB - This study examined both self-rated and objectively measured attractiveness in relation to sexual behaviors and attitudes in an undergraduate sample (N=456). About a quarter of the variance in self-ratings of attractiveness was predicted from combining standard objective measures of attractiveness, including face photo ratings, body mass index, and chest-to-waist ratio for men, and face photo ratings, body mass index, and waist-to-hip ratio for women. Correlations were investigated among self-rated attractiveness, measured attractiveness, the residual component of self-rated attractiveness (controlling for measured attractiveness), and a number of sexual and related variables. Measured attractiveness correlated moderately with sexual behaviors but not with sociosexuality or sexual moral attitudes, indicating that higher levels of observable attractiveness may serve to increase opportunities for sex with multiple desirable partners without affecting interests in or moral acceptance of casual sex. Self-rated attractiveness correlated positively with sexual behaviors and with sociosexuality, but the correlation with sociosexuality was based entirely on residual factors beyond the objective measures of attractiveness. Other predictors of sexual behavior were discussed in terms of their variable roles in affecting interest in, opportunities for, and social costs of promiscuous sexual activity. PMID- 17136592 TI - Associations between religiosity and sexuality in a representative sample of Australian adults. AB - Many studies have examined the influence on sexual attitudes and behavior of religious belief (i.e., religious denomination) or religiosity (e.g., attendance at services, subjective importance of religion). However, few studies have examined the combined effects of religion and religiosity on sexual attitudes and behavior. This study examined such effects in a representative sample of 19,307 Australians aged 16-59 years (response rate, 73.1%). The study compared members of four religious groups (Protestant, Catholic, Buddhist, Muslim) and two levels of frequency of attendance at religious service (less than monthly, at least monthly). Religious participants were compared to their non-religious peers in analyses adjusted for potential confounding by demographic variables. The outcomes were five sexual behaviors and five corresponding measures of sexual attitudes. The study revealed inconsistent patterns of association between religion/religiosity and a range of sexual behaviors and attitudes. In general, greater attendance at religious services was associated with more conservative patterns of behavior and attitudes. However, religious people who attended services infrequently were more similar to their non-religious peers than their more religious peers. The results of this study highlight the importance of considering not only religion or religiosity, but the intersection between these two variables. PMID- 17136593 TI - Localization of latent epileptic activities using spatio-temporal independent component analysis of FMRI data. AB - Localizing interictal epileptic activities is a difficult problem in clinical practice. We report a novel noninvasive technique, resting functional magnetic resonance imaging (fMRI) with spatio-temporal independent component analysis (ICA), for localizing interictal epileptic activities. First, the fMRI data is separated into independent spatial patterns by spatial-ICA, and the patterns with Z-values larger than a threshold are selected as the potential spatial patterns of the epileptic activities. Second, the temporal series of the active points in the selected patterns are separated by temporal-ICA, and the component with the biggest Gaussian deviation (kurtosis) is selected as the representative of the epileptic discharge activity in a sub-region. Finally, those spatial sub-regions, which have distinct epileptic discharge activities confirmed by temporal-ICA are considered as the epileptic foci. This method was applied to fMRI data of six epileptic patients, and the results are consistent with the clinical assessment. Though more studies are required to validate this technique, the above preliminary results demonstrate the potential of using the resting fMRI with spatio-temporal ICA to detect and localize latent epileptic activities. PMID- 17136594 TI - TMS orientation for NIRS-functional motor mapping. AB - Functional near-infrared spectroscopic imaging (NIRS imaging) has the potential to elucidate the relationship between neuronal activity and oxygenation responses. However, its signal specificity to the functional cortex is sometimes spoiled by its rough spatial resolution. In this study we incorporated transcranial magnetic stimulation (TMS) motor mapping into an NIRS imaging study to enhance spatial specificity to the functional cortex. Distinctive biphasic responses in the cortical oxygenation status were observed in the center of the primary motor cortex during a motor task. The early response phase, occurring within 1 to 3 seconds after task initiation, represents a cortical deoxygenation which consists of a significant increase in deoxygenated hemoglobin concentration (HbR) and a nonsignificant decreasing tendency in oxygenated hemoglobin concentration (HbO(2)). The delayed response phase represents an excess of incoming blood flow, which appears as an increase in HbO(2)/total Hb (tHb) and a decrease in HbR following the early response. In the surrounding area, cortical oxygenation change showed a monophasic response consisting of an increase in HbO(2)/tHb and a decrease in HbR. Combining TMS mapping with NIRS imaging enabled us to specify the cortex with the strongest functional activity. PMID- 17136595 TI - Neuronal modifications during visuomotor association learning assessed by electric brain tomography. AB - In everyday life specific situations need specific reactions. Through repetitive practice, such stimulus-response associations can be learned and performed automatically. The aim of the present EEG study was the illustration of learning dependent modifications in neuronal pathways during short-term practice of visuomotor associations. Participants performed a visuomotor association task including four visual stimuli, which should be associated with four keys, learned by trial and error. We assumed that distinct cognitive processes might be dominant during early learning e.g., visual perception and decision making. Advanced learning, however, might be indicated by increased neuronal activation in integration- and memory-related regions. For assessment of learning progress, visual- and movement-related brain potentials were measured and compared between three learning stages (early, intermediate, and late). The results have revealed significant differences between the learning stages during distinct time intervals. Related to visual stimulus presentation, Low Resolution Electromagnetic Brain Tomography (LORETA) revealed strong neuronal activation in a parieto-prefrontal network in time intervals between 100-400 ms post event and during early learning. In relation to the motor response neuronal activation was significantly increased during intermediate compared to early learning. Prior to the motor response (120-360 ms pre event), neuronal activation was detected in the cingulate motor area and the right dorsal premotor cortex. Subsequent to the motor response (68-430 ms post event) there was an increase in neuronal activation in visuomotor- and memory-related areas including parietal cortex, SMA, premotor, dorsolateral prefrontal, and parahippocampal cortex. The present study has shown specific time elements of a visuomotor-memory-related network, which might support learning progress during visuomotor association learning. PMID- 17136596 TI - Elastic and rupture properties of porcine aortic tissue measured using inflation testing. AB - A new inflation test device was developed to study the mechanical properties of aortic tissue. The device was used to measure failure (rupture) strength and to determine the nonlinear, anisotropic elastic properties of porcine thoracic aorta. The tester was designed to stretch initially flat, circular tissue specimens to rupture under uniform biaxial loading. Water was chosen as the pressurizing fluid. Mechanical stretch and radius of curvature during inflation were measured optically in two orthogonal directions, and the Cauchy stress components were calculated from the deformation and the applied pressure. All porcine samples that ruptured successfully did so via a tear in the circumferential direction. Thus, the failure strength was taken to be the stress in the axial direction immediately prior to rupture. The mean failure strength was 1.75 MPa and mean axial stretch at failure was 1.52. These values agree well with published data for other arterial tissues. The nonlinearly elastic deformation behavior was modeled using a hyperelastic constitutive law of the type proposed by Holzapfel et al. [Holzapfel GA, Gasser TC, Ogden RW. J Elasticity 2000;61:1-48]. The results showed that the dominant directions of anisotropy in the porcine aortas were approximately 45 degrees to the axial and circumferential directions, and that the isotropic contribution to the constitutive model was insignificant. PMID- 17136597 TI - Construction of an artificial heart pump performance test system. AB - A hydraulic loop, which simulates pressure/flow response of the human circulatory system, is needed to bench test the various versions of rotary left ventricular assist devices (LVADs). This article describes the design of such a loop and the simulated response of different physiological states, such as a healthy person in sleep, rest, and mild physical activity, and in different pathological states. The loop consists of: (1) pulsatile left and right cardiac simulators; (2) air/water tanks to model the venous and arterial compliances; (3) tygon tubes to model the venous, arterial, and other system flow resistances; and (4) a tuning clamp to model the variation in system resistance characteristics under different cardiac pressure/flow conditions. The simulated responses were compared to the data found in the literature to validate the loop performance prior to LVAD testing. PMID- 17136598 TI - Differential expression, time course and distribution of four PARs in rats with endotoxin-induced acute lung injury. AB - The hypothesis that the expression of protease-activated receptors (PARs) protein is regulated at the level of transcription and that PAR isoforms, PAR-1, PAR-2, PAR-3, and PAR-4, in lung tissue show different patterns of expression in lipopolysaccharide (LPS)-induced acute lung injury (ALI) was tested. Male Wistar rats were rendered endotoxemic by intra-peritoneal injection of LPS (15 mg/kg body weight). We examined the expression of protein and mRNA and the immunohistochemical localization of PAR isoforms in lung tissues 1, 3, 6, and 10 h after LPS administration. Induction of ALI by LPS was confirmed based on histopathological changes. LPS administration induced significant increases in the expression of PAR isoforms (protein) at the level of transcription in ALI. While the time course of PAR-1 and -2 expressions were different, those of PAR-3 and -4 were almost similar. An immunohistochemical analysis showed localization of PAR isoforms in the vascular endothelium, alveolar epithelium, and alveolar macrophages. However, the cellular distribution patterns of PAR isoforms were different. We conclude that LPS induces increase in protein expression of PAR isoforms at the level of transcription in rats with ALI. The differential expression patterns (over a time course) and distribution of PAR isoforms suggests a distinct role for each isoform in the pathogenesis of LPS-induced ALI. PMID- 17136599 TI - Severity of illness and the welfare effects of moral hazard. AB - The extent to which the moral hazard caused by health insurance represents economic inefficiency has been the subject of much debate. This paper incorporates health status in a model of moral hazard, and finds that seriously ill patients are likely to exhibit greater moral hazard than healthier patients but the proportion of moral hazard that is inefficient declines with the severity of illness. Because of these competing tendencies, the cost of resource misallocation is parabolic in the severity of illness. The effect of the consumer's initial wealth endowment is also considered. PMID- 17136600 TI - A tribute to Eric Schopler. PMID- 17136601 TI - Addressing the idiosyncratic needs of Orthodox Jewish couples requesting sex selection by preimplantation genetic diagnosis (PGD). AB - We report here on ethical considerations addressing the idiosyncratic needs of two Orthodox Jewish couples requesting sex selection through PGD. The patients' considerations stem from generally healthy concerns, are not based on any gender biases and have little chance of having any major societal impact, given the idiosyncratic nature of the situation. Halakhah, the legal and ethical system of rabbinic Orthodox Judaism, generally opposes sex selection through PGD for nonmedical reasons, but would approve the procedure in these cases. Meeting these needs within the context of the doctor-patient relationship necessitates reconsidering to some extent the ASRM Ethics Committee guidelines. PMID- 17136602 TI - Hysteresis of a biomaterial: influence of sutures and biological adhesives. AB - We studied the changes in energy consumption of samples of calf pericardium, when joined or not joined by sutures and adhesives, by means of hysteretic cycles. Sixty-four samples were subsequently subjected to tensile stress until rupture. An overlapping suture sewn in the form of a rectangle presented an acceptable mean resistance to rupture of over 10 MPa, although lower than the mean values in an unsutured control series where the mean resistance surpassed 15 MPa. The contribution of an acrylic adhesive to the resistance to rupture was negligible. The sutured samples that were reinforced with adhesives and had not been subjected to hysteretic cycles prior to rupture showed an anisotropic behavior. This behavior appeared to be lost in all the samples that underwent hysteretic cycles. We found an inflection point in the stress/strain curve following the stepwise increase in the load, with a value greater than and proximate to the final load applied. This inflection should be analyzed by means of microscopy. Finally, the mathematical relationship between the energy consumed and the stress applied, the strain or deformation produced and the number of cycles of hysteresis to which the samples were subjected was established as the ultimate objective of this study. The bonding systems provoked a greater consumption of energy, with the greatest consumption corresponding to the first cycle in all the series assayed. An equation relating the energy consumption in a sample to the number of hysteretic cycles to which it was subjected was obtained. Its asymptote on the x-axis indicates the energy consumption for a theoretical number of cycles, making it possible to estimate the durability of the sample. PMID- 17136603 TI - Effect of gamma-irradiation on the structure of poly(ethyl acrylate-co hydroxyethyl methacrylate) copolymer networks for biomedical applications. AB - The effect of the sterilization process by gamma-irradiation on the structure of poly(ethyl acrylate-co-hydroxyethyl methacrylate) copolymer networks, P(EA-co HEMA) is studied for a broad dose range (7, 15, 25 and 50 kGy) and copolymer composition interval (0, 0.3, 0.5, 0.7 and 1 weight fraction of HEMA in the copolymer). gamma-irradiation promotes chain scission in PHEMA homopolymer but induces new crosslinking points in PEA homopolymer. Both effects are present in the copolymers, with a net result that depends on composition. For copolymers with high HEMA fractions chain scission predominates, while, as the amount of EA in the copolymer increases, the situation changes and the net effect turns out to be an increase in the number of elastically active chains. Further, gamma irradiation strengthens the gamma relaxation in PHEMA homopolymer, what suggest that the number of interchain hydrogen bonds decreases. FTIR spectroscopy reveals no oxidation as a consequence of the sterilization process. PMID- 17136604 TI - Sonochemical synthesis of calcium phosphate powders. AB - beta-tricalcium phosphate (beta-TCP) and biphasic calcium phosphate powders (BCP), consisting of hydroxyapatite (HA) and beta-TCP, were synthesized by thermal decomposition of precursor powders obtained from neutralization method. The precursor powders with a Ca/P molar ratio of 1.5 were prepared by adding an orthophosphoric acid (H(3)PO(4)) solution to an aqueous suspension containing calcium hydroxide (Ca(OH)(2)). Mixing was carried out by vigorous stirring and under sonochemical irradiation at 50 kHz, respectively. Glycerol and D-glucose were added to evaluate their influence on the precipitation of the resulting calcium phosphate powders. After calcination at 1000 degrees C for 3 h BCP nanopowders of various HA/beta-TCP ratio were obtained. PMID- 17136605 TI - Effect of calcium, zinc and magnesium on the attachment and spreading of osteoblast like cells onto ceramic matrices. AB - Calcium phosphate ceramic has been widely used as bone substitute materials. Neumerours approaches have been investigated to develop tissue-engineered scaffold from hydroxyapatite because of its advantages like osteoconduction. We have developed porous ceramic matrices from nanoparticles of calcium phosphate containing zinc and magnesium. Mimicking the grain size of natural bone enhances the bone forming function of cells. Osteoblast-like MG63 cells were cultured on to these porous ceramic matrices. Cell adhesion and spreading onto these matrices were studied for 24 h and 5 days in vitro. It was observed that on calcium phosphate matrix, containing a combination of zinc and magnesium, the osteoblast adhesion and spreading was significant on 5th day. This appeared to be comparable to the hydroxyapatite control. This makes it a promising candidate as a bone tissue-engineering scaffold. PMID- 17136606 TI - Raman spectroscopy of the human nail: a potential tool for evaluating bone health? AB - Dual X-ray absorptiometry (DEXA) is the current gold standard for the diagnosis of osteoporosis. However, patients can suffer osteoporotic fractures despite normal bone mineral density, partly because of unmeasured influences of both the protein and mineral phases of bone that are affected in osteoporosis. There is currently no clinically applicable method of evaluating the health of the protein phase. The proteins in human nail (keratin) and bone (collagen) require sulphation and disulphide bond (S-S) formation for structural integrity and disorders of either sulphur metabolism or cystathione beta-synthase can lead to structural abnormalities in these tissues. Raman protein spectra provide a method of non-invasive measurement of the degree of sulphation of structurally related proteins that may be indicative of bone health. Raman spectroscopy was used to evaluate the disulphide (S-S) content of fingernails. The nail samples came from from 169 women (84 pre- and 85 post-menopausal), of which 39 had a history of osteoporotic fracture. BMD was measured by DXA at the spine. Analyses included parametric and non-parametric tests, dependent on the distribution of the test variable. Mean disulphide content of the nail reduced with age and was slightly higher in pre-, compared to post-menopausal women (P = 0.187). Significantly lower disulphide content was observed in nails obtained from subjects with a history of fracture (P = 0.025). When either disulphide content or BMD was used as a predictor, the odds ratio of these two measures were found to be comparable predictors for fracture status. This suggests that measurements of change in the protein phase of structural proteins such as keratin in the human nail may be correlated with clinically relevant changes in bone proteins that are important in fracture risk. PMID- 17136607 TI - Preparation and characterization of pH sensitive comb-shaped chitosan material for the controlled release of coenzyme A. AB - A novel kind of pH sensitive comb-shaped copolymer P(CS-Ma-PEGMA) was synthesized with chitosan (CS), maleic anhydride (Ma) and Poly (ethylene glycol) methacrylate (PEGMA) by grafting and co-polymerization. The structure of P(CS-Ma-PEGMA) was characterized by FT-IR and (1)H-NMR, and it was found that PEGMA was grafted onto CS and PEGMAylated chitosan was soluble. The copolymer was subjected to coenzyme A adsorption study in order to assess its application in biomedical area. The factors affecting release behavior, such as concentration and pH were discussed in this paper. The higher concentration of the copolymer showed higher absorbance peak than the lower one. The pH of the solution also had significant impact on the release of coenzyme A, and the mechanism of adsorption was suggested. The results suggested that the novel copolymer could be used as drug delivery carrier. PMID- 17136608 TI - Osteogenic behavior of alginate encapsulated bone marrow stromal cells: an in vitro study. AB - Sodium alginate is a useful polymer for the encapsulation and immobilization of a variety of cells in tissue engineering because it is biocompatible, biodegradable and easy to process into injectable microbeads. Despite these properties, little is known of the efficacy of calcium cross-linked alginate gel beads as a biodegradable scaffold for osteogenic cell proliferation and differentiation. In this study, we investigated the ability of rabbit derived bone marrow cells (BMCs) to proliferate and differentiate in alginate microbeads and compared them with BMCs cultured in poly-L-lysine (PLL) coated microbeads and on conventional 2D plastic surfaces. Results show that levels of proliferation and differentiation in microbeads and on tissue culture plastics were comparable. Cell proliferation in microbeads however diminished after fortification with a coating layer of PLL. Maximum cell numbers observed were, 3.32 x 10(5) +/- 1.72 x 103; 3.11 x 10(5) +/- 1.52 x 10(3) and 3.28 x 10(5) +/- 1.21 x 10(3 ) for the uncoated, PLL coated and plastic surface groups respectively. Alkaline phosphatase and protein expressions reflected the stage of cell differentiation. We conclude that calcium cross-linked alginate microbeads can act as a scaffold for BMC proliferation and osteogenic differentiation and has potential for use as 3D degradable scaffold. PMID- 17136609 TI - Combined effects of staining substances on resin composites before and after surface sealant application. AB - The objective was to measure the combined effect of mucin, chlorhexidine and tea solution on the staining of four dental resin composites, and to determine the effect of surface sealant on staining. One side of cured resin composite specimens of 10 mm in diameter and 2 mm in thickness were polished with 600-grit silicon carbide paper. One group of specimens (n = 5) was treated with a surface sealant [BisCover, Bisco, USA; SS (surface sealant) group], and the other group was not (NO group; control). Specimens were sequentially immersed in the following substances: Mucin in phosphate buffered saline (PBS); chlorhexidine; tea solution; and ultrasonic cleaning and then immersion in PBS. Color was measured on a reflection spectrophotometer. Changes in color (DeltaE (*) (ab)) and color parameters, such as hue, chroma and value, after immersion in tea solution and subsequent cleaning were analyzed by repeated measures, analysis of variance at the 0.05 level of significance. The range of DeltaE (*) (ab) values after immersion in tea solution was 11.4-21.1 for NO group and 10.5-19.6 for SS group, and that after cleaning was 2.4-10.0 for NO group and 2.7-8.3 for SS group. After staining, CIE L (*) value (lightness) decreased, and CIE a (*) and b (*) values increased. Color changes of resin composites were not acceptable after sequential immersion treatment (DeltaE (*) ( ab ) > 3.3). The changes in color and color parameters of sealant applied group were not significantly different from those of control group except for a few combinations of color parameters and resin composites. PMID- 17136610 TI - Carbohydrate restriction does not change mitochondrial free radical generation and oxidative DNA damage. AB - Many previous investigations have consistently reported that caloric restriction (40%), which increases maximum longevity, decreases mitochondrial reactive species (ROS) generation and oxidative damage to mitochondrial DNA (mtDNA) in laboratory rodents. These decreases take place in rat liver after only seven weeks of caloric restriction. Moreover, it has been found that seven weeks of 40% protein restriction, independently of caloric restriction, also decrease these two parameters, whereas they are not changed after seven weeks of 40% lipid restriction. This is interesting since it is known that protein restriction can extend longevity in rodents, whereas lipid restriction does not have such effect. However, before concluding that the ameliorating effects of caloric restriction on mitochondrial oxidative stress are due to restriction in protein intake, studies on the third energetic component of the diet, carbohydrates, are needed. In the present study, using semipurified diets, the carbohydrate ingestion of male Wistar rats was decreased by 40% below controls without changing the level of intake of the other dietary components. After seven weeks of treatment the liver mitochondria of the carbohydrate restricted animals did not show changes in the rate of mitochondrial ROS production, mitochondrial oxygen consumption or percent free radical leak with any substrate (complex I- or complex II-linked) studied. In agreement with this, the levels of oxidative damage in hepatic mtDNA and nuclear DNA were not modified in carbohydrate restricted animals. Oxidative damage in mtDNA was one order of magnitude higher than that in nuclear DNA in both dietary groups. These results, together with previous ones, discard lipids and carbohydrates, and indicate that the lowered ingestion of dietary proteins is responsible for the decrease in mitochondrial ROS production and oxidative damage in mtDNA that occurs during caloric restriction. PMID- 17136611 TI - Welfare reform and health of immigrant women and their children. AB - We investigate the association between the 1996 welfare reform and health insurance, medical care use and health of low-educated, foreign-born, single mothers and their children. We find that welfare reform was associated with an eight to 11.5 percentage points increase in proportion uninsured among low educated foreign-born, single mothers. We also find that the decline in welfare caseload since 1996 was associated with a 6.5 to 10 percentage points increase in the proportion of low-educated foreign-born, single mothers reporting delays in receiving medical care or receiving no care due to cost and a nine percentage points decline in visits to a health professional in the past 12 months. We do not find any consistent evidence that welfare reform affected the health insurance, medical care utilization and health of children living with single mothers. PMID- 17136612 TI - Identifying potential risk and protective factors among non-metropolitan Latino youth: cultural implications for substance use research. AB - Immigration studies show that the social adaptation of second-generation youth is conditioned by the pace of acculturation among parents and children, cultural and economic barriers, and family and community resources for confronting barriers. This research, however, has primarily focused on the link between acculturation and acculturative stress on Latino adolescents residing in large urban communities. There is a lack of research on the social integration of Latino youth living in rapidly expanding non-metropolitan communities. Consequently, we explored cultural aspects and potential risk and protective factors for early onset of alcohol use for Latino youth. Our findings indicate these rural Latino youth face unique and common stressors compared to urban youth that place them at risk for alcohol use. Cultural expectations surrounding substance use, however, may serve as protective factors to substance use for Latino youth, particularly girls. PMID- 17136613 TI - Secretion and fluid transport mechanisms in the mammary gland: comparisons with the exocrine pancreas and the salivary gland. AB - Milk is a complex fluid composed of proteins, sugars, lipids and minerals, in addition to a wide variety of bioactive molecules including vitamins, trace elements and growth factors. The composition of these components reflects the integrated activities of distinct synthetic, secretion and transport processes found in mammary epithelial cells, and mirrors the differing nutritional and developmental requirements of mammalian neonates. Five general pathways have been described for secretion of milk components. With the exception of lipids, which are secreted a unique pathway, milk components are thought to be secreted by adaptations of pathways found in other secretory organs. However little is known about the molecular and cellular mechanisms that constitute these pathways or the physiological mechanisms by which they are regulated. Comparisons of current secretion and transport models in the mammary gland, exocrine pancreas and salivary gland indicate that significant differences exist between the mammary gland and other exocrine organs in how proteins and lipids are packaged and secreted, and how fluid is transported. PMID- 17136614 TI - Analysis of lactation defects in transgenic mice. AB - Although lactation is the only physiological function of the mammary gland, little is known about the molecular events required for secretory activation and milk production. Genetically altered mice have been used extensively to study mammary gland development during puberty and pregnancy, as well as mammary tumorigenesis. A number of approaches have been used to produce genetic modifications in mammary glands of mice, including transgenic mice utilizing mammary specific promoters, traditional knockout mice, mammary-specific gene deletion, and conditionally-regulated transgenes. The same technologies can be used to study secretory activation and lactation; however only a comparatively small number of studies to date have used these approaches to study these events. In this paper we review the technologies available to make genetically modified mice for the study of secretory activation and lactation as well as specific analytical procedures that can be used to characterize mice with lactation defects. PMID- 17136615 TI - Workplace injury management: using new technology to deliver and evaluate physician continuing medical education. AB - OBJECTIVE: Physicians typically receive little continuing medical education (CME) about their role in workplace injury management as well as on workplace injuries and disease. Although new technologies may help educate physicians in these areas, careful evaluation is required, given the understudied nature of these interventions. The objective of this study is to evaluate two promising new technologies to deliver CME (online learning and videoconferencing) and to compare the effectiveness of these delivery methods to traditional CME interventions (large urban traditional conference lectures and small group local face-to-face outreach) in their impact on physician knowledge related to workplace injury management. METHODS: This study utilized a prospective, controlled evaluation of two educational programs for BC physicians: 1) The Diagnosis and Management of Lateral Epicondylitis; and 2) Is Return-to-Work Good Medicine? Each educational module was delivered in each of four ways (Outreach Visit, Videoconference Session, Conference Lecture, Online) and physicians self selected their participation--both in terms of topic and delivery method. Questionnaires related to knowledge as well as learner attitude and satisfaction were administered prior (pre-test) and following (post-test) all educational sessions. RESULTS: 581 physician encounters occurred as a result of the educational interventions and a significant percentage of the physicians participated in the research per se (i.e. there were 358 completed sets of pre test and post-test 'Knowledge' questionnaires). Overall the results showed that the developed training programs increased physicians' knowledge of both Lateral Epicondylitis and the physician's role in Return-To-Work planning as reflected in improved post-test performance when compared to pre-test scores. Furthermore, videoconferencing and online training were at least as effective as conference lectures and instructor-led small group outreach sessions in their impact on physician knowledge. CONCLUSIONS: Use of effective videoconferencing and online learning activities will increase physician access to quality CME related to workplace injury management and will overcome access barriers intrinsic to types of CME interventions based on instructor-student face-to-face interactions. PMID- 17136616 TI - Human eosinophil major basic protein 2: location of disulfide bonds and free sulfhydryl groups. AB - Eosinophil granule major basic protein 2 (MBP2 or major basic protein homolog) is a paralog of major basic protein (MBP1) and, similar to MBP1, is cytotoxic and cytostimulatory in vitro. MBP2, a small protein of 13,433 Da molecular weight, contains 10 cysteine residues. Mass spectrometry shows two cystine disulfide linkages (Cys20-Cys115 and Cys92-Cys107) and 6 cysteine residues with free sulfhydryl groups (Cys2, Cys23, Cys42, Cys43, Cys68, and Cys96). MBP2, similar to MBP1, has conserved motifs in common with C-type lectins. The disulfide bond locations are conserved among human MBP1, MBP2 and C-type lectins. PMID- 17136617 TI - The use of the free metal-temperature 'phase diagrams' for studies of single site metal binding proteins. AB - Typical physico-chemical studies of metal binding proteins are usually aimed at determination of the metal binding constant K for a native protein (Kn), while the significance of the K value for the thermally denatured protein (Ku) is usually underestimated. Meanwhile, metal binding induced shift of thermal denaturation transition of a single site metal binding protein is defined by Kn to Ku ratio, implying that knowledge of both K values is required for full characterization of the system. In the present work, the most universal approach to the studies of single site metal binding proteins, namely construction of a protein "phase diagram" in coordinates of free metal ion concentration - temperature, is considered in detail. The detailed algorithm of construction of the phase diagrams along with underlying mathematic procedures developed here may be of use for studies of other simple protein-target type systems, where target represents low molecular weight ligand. Analysis of the simplest protein-ligand system reveals that thermodynamic properties of apo-protein dictate the maximal possible increase of its affinity to any simple ligand upon thermal denaturation of the protein. Experimental and general problems coupled with the use of the phase diagrams are discussed. PMID- 17136618 TI - George Albee, exemplar of primary prevention. PMID- 17136619 TI - In gratitude to George W. Albee, prevention's true north. PMID- 17136620 TI - In memory of George Albee. PMID- 17136621 TI - Bayesian dynamic models for survival data with a cure fraction. AB - In this paper, we propose a new class of semi-parametric cure rate models. Specifically, we construct dynamic models for piecewise hazard functions over a finite partition of the time axis. Allowing the size of partition and the levels of baseline hazard to be random, our proposed models provide a great flexibility in controlling the degree of parametricity in the right tail of the survival distribution and the amount of correlations among the log-baseline hazard levels. Several properties of the proposed models are derived, and propriety of the implied posteriors with improper noninformative priors for regression coefficients based on the proposed models is established for the fixed partition of the time axis. In addition, an efficient reversible jump computational algorithm is developed for carrying out posterior computation. A real data set from a melanoma clinical trial is analyzed in detail to further demonstrate the proposed methodology. PMID- 17136622 TI - Functional abnormalities of the motor tract in the rat after portocaval anastomosis and after carbon tetrachloride induction of cirrhosis. AB - INTRODUCTION: Hepatic encephalopathy is a neurologic syndrome secondary to liver failure that causes cognitive and motor abnormalities. Impairment in the function of the first neuron of the motor tract (corticospinal tract) has been demonstrated in patients with cirrhosis and minimal hepatic encephalopathy. AIM: Investigate the function of the first neuron of the motor tract in experimental models of minimal hepatic encephalopathy. MATERIAL AND METHODS: Rats with portocaval anastomosis (n = 8) and rats with carbon tetrachloride induced cirrhosis (n = 11) underwent neurophysiological recording under light anesthesia with propofol. Motor evoked potentials were elicited applying a transcranial electric pulse and were recorded in the tibialis anterior muscle. The effect of the dose of anesthesia was assessed in a group of normal rats (n = 10). RESULTS: Rats with portocaval anastomosis exhibited a decrease in motor evoked potentials amplitude following surgery (67 +/- 11 to 41 +/- 16%, P < 0.001). Cirrhotic rats exhibited an increase in motor evoked potentials latency after the appearance of ascites (4.65 +/- 0.43 to 5.15 +/- 0.67 ms., P = 0.04). Increasing doses of propofol produced a decrease in the amplitude and an increase in the latency of motor evoked potentials. CONCLUSION: It is possible to reproduce functional abnormalities of the central motor tract in rats with portocaval anastomosis and carbon tetrachloride induced cirrhosis. The development of motor abnormalities in experimental models of minimal hepatic encephalopathy offers the possibility to investigate the mechanisms involved in the pathogenesis of hepatic encephalopathy and test therapeutic strategies. PMID- 17136623 TI - The effectiveness of sexual offender treatment for juveniles as measured by recidivism: a meta-analysis. AB - Published and unpublished data from nine studies on juvenile sexual offender treatment effectiveness were summarized by meta-analysis (N=2986, 2604 known male). Recidivism rates for sexual, non-sexual violent, non-sexual non-violent crimes, and unspecified non-sexual were as follows: 12.53%, 24.73%, 28.51%, and 20.40%, respectively, based on an average 59-month follow-up period. Four included studies contained a control group (n=2288) and five studies included a comparison treatment group (n=698). An average weighted effect size of 0.43 (CI=0.33-0.55) was obtained, indicating a statistically significant effect of treatment on sexual recidivism. However, individual study characteristics (e.g., handling of dropouts and non-equivalent follow-up periods between treatment groups) suggest that results should be interpreted with caution. A comparison of odds ratios by quality of study design indicated that higher quality designs yielded better effect sizes, though the difference between groups was not significant. PMID- 17136624 TI - A comparison of the application of the self-regulation model of the relapse process for mainstream and special needs sexual offenders. AB - The self-regulation model of the relapse process (Ward & Hudson, 2000) has been developed and empirically validated on general sexual offender populations (Bickley & Beech, 2002), but not on specific sexual offender populations. This paper aims to investigate whether special needs offenders, as compared to mainstream sexual offenders, can be categorized into the offense pathways described in the model. In addition, this paper aims to evaluate the application of the self-regulation model in highlighting the treatment needs of the special needs group. Special needs sexual offenders are defined as a treatment population that includes individuals with lower functioning, limited social and communication skills, and literacy deficits. Participants were classified into the self-regulation model using a method developed by Bickley and Beech (2002). Demographic and offense information were collected and comparisons made between the special needs and mainstream groups. The results showed that the sexual offenders with special needs could be reliably classified into the offense pathways of the self-regulation model. The largest group of special needs offenders was in the approach-automatic group, followed by the approach-explicit group. The results indicated no significant differences in representation in the offense pathways between the special needs and mainstream sexual offenders. The results also indicate that the special needs group would benefit from a responsive approach to treatment, which incorporates appropriate treatment targets identified by the self-regulation model. PMID- 17136625 TI - Different actuarial risk measures produce different risk rankings for sexual offenders. AB - Percentile ranks were computed for N=262 sex offenders using each of 5 actuarial risk instruments commonly used with adult sex offenders (RRASOR, Static-99, VRAG, SORAG, and MnSOST-R). Mean differences between percentile ranks obtained by different actuarial measures were found to vary inversely with the correlation between the actuarial scores. Following studies of factor analyses of actuarial items, we argue that the discrepancies among actuarial instruments can be substantially accounted for by the way in which the factor Antisocial Behavior and various factors reflecting sexual deviance are represented among the items contained in each instrument. In the discussion, we provide guidance to clinicians in resolving discrepancies between instruments and we discuss implications for future developments in sex offender risk assessment. PMID- 17136626 TI - Risk factors for adolescent sex offender recidivism: evaluation of predictive factors and comparison of three groups based upon victim type. AB - This study investigated differences in recidivism risk factors and traits associated with psychopathy among 3 groups of male adolescent sexual offenders (N=156): offenders against children, offenders against peers or adults, and mixed type offenders. Furthermore, those same variables were examined for their association with sexual and nonsexual recidivism and the 3 groups were compared for differences in rates of recidivism. Based upon both juvenile and adult recidivism data, 6.4% of the sample reoffended sexually and 30.1% reoffended nonsexually. Retrospective risk assessments were completed using the Juvenile Sex Offender Assessment Protocol-II (JSOAP-II) and the Psychopathy Checklist:Youth Version (PCL:YV). Comparisons of the 3 preexisting groups for differences on scale and factor scores were conducted using analyses of variance (ANOVAs). Differences among groups for recidivism were measured using survival curve analysis. Associations between risk scales and recidivism were measured using Cox regression analyses. Results suggest significant differences among the 3 offender groups on multiple scales of the JSOAP-II and PCL:YV, with mixed type offenders consistently producing higher risk scores as compared to those who exclusively offend against children or peers/adults. The Impulsive/Antisocial Behavior scale of the JSOAP-II and the Interpersonal and Antisocial factors of the PCL:YV were significant predictors of sexual recidivism. The Behavioral and Antisocial factors of the PCL:YV were significant predictors of nonsexual recidivism. Results supported previous research indicating that most adolescents who sexually offend do not continue offending into adulthood. Such results can lead to improved treatment by targeting specific risk factors for intervention and better use of risk management resources in the community, while preserving the most restrictive treatment options for the highest risk offenders. PMID- 17136627 TI - Does Static-99 predict recidivism among older sexual offenders? AB - Static-99 (Hanson & Thornton, 2000) is the most commonly used actuarial risk tool for estimating sexual offender recidivism risk. Recent research has suggested that its methods of accounting for the offenders' ages may be insufficient to capture declines in recidivism risk associated with advanced age. Using data from 8 samples (combined size of 3,425 sexual offenders), the present study found that older offenders had lower Static-99 scores than younger offenders and that Static 99 was moderately accurate in estimating relative recidivism risk in all age groups. Older offenders, however, had lower sexual recidivism rates than would be expected based on their Static-99 risk categories. Consequently, evaluators using Static-99 should considered advanced age in their overall estimate of risk. PMID- 17136628 TI - Adolescent sexual offenders: a total survey of referrals to Social Services in Sweden and subgroup characteristics. AB - Sampling methodology (e.g. population-based vs. clinical samples, anonymous self reports vs. data collected as part of mandated treatment) affects the validity of conclusions drawn from research addressing the etiology of adolescent sexual offending. Studies of unselected samples allow testing of the generalizability of etiological models suggested from investigation of selected clinical or forensic populations. Further, representative epidemiological data on adolescent sexual offending is needed for policy-making and the planning of services. We conducted a national survey of all adolescent sexual offenders (ASOs, 12-17 years) referred to Social Services during 2000. Social workers at all child and adolescent units in Social Service authorities throughout Sweden (N=285, 99% response rate) completed a questionnaire about new ASO referrals in 2000. The National Board of Health and Welfare commissioned the survey and questionnaire items tapped offender, offence, and victim characteristics. A total of 197 boys and 2 girls aged 12-17 years were referred to Social Services because of sexually abusive behavior in 2000. Focusing specifically on males, this yielded a one-year incidence of .060% (95% confidence interval = .052-.068). Forty-six percent of male ASOs abused at least one child younger than age 12 years (child offenders) whereas the rest had abused peer or adult victims (peer offenders). Forty-two percent of male ASOs had ever committed sexual offences together with at least one other offender (group offenders). Child- vs. peer offenders and group vs. single offenders, suggested typologies in the literature, were compared to explore potential subtype-specific risk factors and correlates. The results suggested a higher proportion of group ASOs than previously reported and stronger support for subdividing ASOs into child vs. peer offenders than into group vs. single ASOs. PMID- 17136629 TI - Deaths in the Cook County jail: 10-year report, 1995-2004. AB - The aims of this study were to describe causes of death during the 10-year period between 1995 and 2004 in a large urban jail in Chicago; to compare disease specific mortality rates between the jail population and the general population; to explore demographic and incarceration characteristics of the inmates who died in the jail by cause of death; and to examine gender difference in demographic characteristics, incarceration patterns, and causes of death. A total of 178 deaths occurring in the jail over a 10-year period (1995-2004) were reviewed. Age adjusted disease-specific mortality rates were computed for the jail population and compared with the rates in the US general population. Cause of death, demographic variables, and incarceration related factors were retrieved from multiple computerized databases. Descriptive analyses were performed to examine demographic and incarceration-related patterns by cause of death and gender. Heart disease was the most frequent cause of death in the jail population, followed by cerebrovascular disease and suicide. Mortality rates for heart diseases, infectious/inflammatory conditions and suicide were higher for jail inmates than the general population. Black inmates accounted for the majority of deaths due to illnesses and homicide, and a much higher proportion of white and Hispanic inmates were involved in suicide deaths. Deaths due to drug overdose or withdrawal were disproportionately higher among female inmates compared with male inmates. Consistent review of mortality rates and causes of deaths in jail can be a useful tool to better understand health issues and needs of jail inmates. Surveillance of acute and chronic illnesses and strategic reengineering of jail health care is a key to quality improvement for incarcerated populations for whom the jail system becomes their primary care provider. PMID- 17136630 TI - A comparison of spatial and spectral image resolution for mapping invasive plants in coastal california. AB - We explored the potential of detecting three target invasive species: iceplant (Carpobrotus edulis), jubata grass (Cortaderia jubata), and blue gum (Eucalyptus globulus) at Vandenberg Air Force Base, California. We compared the accuracy of mapping six communities (intact coastal scrub, iceplant invaded coastal scrub, iceplant invaded chaparral, jubata grass invaded chaparral, blue gum invaded chaparral, and intact chaparral) using four images with different combinations of spatial and spectral resolution: hyperspectral AVIRIS imagery (174 wavebands, 4 m spatial resolution), spatially degraded AVIRIS (174 bands, 30 m), spectrally degraded AVIRIS (6 bands, 4 m), and both spatially and spectrally degraded AVIRIS (6 bands, 30 m, i.e., simulated Landsat ETM data). Overall success rates for classifying the six classes was 75% (kappa 0.7) using full resolution AVIRIS, 58% (kappa 0.5) for the spatially degraded AVIRIS, 42% (kappa 0.3) for the spectrally degraded AVIRIS, and 37% (kappa 0.3) for the spatially and spectrally degraded AVIRIS. A true Landsat ETM image was also classified to illustrate that the results from the simulated ETM data were representative, which provided an accuracy of 50% (kappa 0.4). Mapping accuracies using different resolution images are evaluated in the context of community heterogeneity (species richness, diversity, and percent species cover). Findings illustrate that higher mapping accuracies are achieved with images possessing high spectral resolution, thus capturing information across the visible and reflected infrared solar spectrum. Understanding the tradeoffs in spectral and spatial resolution can assist land managers in deciding the most appropriate imagery with respect to target invasives and community characteristics. PMID- 17136631 TI - Technology penetration of endovascular aortic aneurysm repair in southern California. AB - Our objective was to investigate the penetration of endovascular abdominal aortic aneurysm repair (EVAR) in the large, diverse health-care market of southern California over 3 years and to study variability in the pattern of distribution of EVAR in southern California counties by analyzing available demographic, geographic, and socioeconomic data from California state health-care databases. Information abstracted from the inpatient hospital discharge data for patients undergoing AAA repair for the years 2001, 2002, and 2003, derived from the Office of Statewide Health Planning and Development, included age, gender, race, hospitals performing EVAR, and payors for the service. Per-capita income (PCI) for the year 1999 and the population size of each county for the respective years were obtained from the U.S. Census Bureau. Data pertaining to members of the Southern California Vascular Surgical Society (SCVSS) serving the southern California region were obtained from the SCVSS membership directory. Data were categorized based on 10 counties in southern California. All the above variables were analyzed using the chi-squared test, with p < 0.05 considered significant. The proportions of EVAR for the years 2001, 2002, and 2003 were 15.4% (n = 409), 20.2% (n = 492), and 25.9% (n = 566), respectively. This is a 67.8% (p < 0.0001) increase in EVAR application in southern California since 2001. However, the proportion of EVAR varied among counties (p < 0.0001), with 457 EVARs performed in Los Angeles County and eight in Imperial County during the study period. EVAR proportion was higher in patients aged > or =65 years (p < 0.0001) and male patients (p < 0.0001). The proportion of EVAR was significantly higher in counties with more than 20 vascular surgeons available (p < 0.0001) and PCI >21,000 US$ (p < 0.0001) and in Medicare, health maintenance organization, preferred provider organization, and private insurance holders (p < 0.0001). There was a trend toward increased EVARs in counties with more than eight hospitals that performed EVAR (p = 0.0545). However, no significant difference in EVAR proportion was observed among subgroups based on race (p = 0.535) and population size (p = 0.84). Although the number and proportion of EVAR increased significantly in southern California over 3 years, the penetration of the procedure varied among counties. County affluence, payor mix, and the number of vascular surgeons/county influenced the variability. These observations suggest that economic barriers may limit access to new biomedical technology. This has implications for health-care public policy directed toward providing equal access to medical care without regard to economic status. PMID- 17136632 TI - Audiological evaluation of affected members from a Dutch DFNA8/12 (TECTA) family. AB - In DFNA8/12, an autosomal dominantly inherited type of nonsyndromic hearing impairment, the TECTA gene mutation causes a defect in the structure of the tectorial membrane in the inner ear. Because DFNA8/12 affects the tectorial membrane, patients with DFNA8/12 may show specific audiometric characteristics. In this study, five selected members of a Dutch DFNA8/12 family with a TECTA sensorineural hearing impairment were evaluated with pure-tone audiometry, loudness scaling, speech perception in quiet and noise, difference limen for frequency, acoustic reflexes, otoacoustic emissions, and gap detection. Four out of five subjects showed an elevation of pure-tone thresholds, acoustic reflex thresholds, and loudness discomfort levels. Loudness growth curves are parallel to those found in normal-hearing individuals. Suprathreshold measures such as difference limen for frequency modulated pure tones, gap detection, and particularly speech perception in noise are within the normal range. Distortion otoacoustic emissions are present at the higher stimulus level. These results are similar to those previously obtained from a Dutch DFNA13 family with midfrequency sensorineural hearing impairment. It seems that a defect in the tectorial membrane results primarily in an attenuation of sound, whereas suprathreshold measures, such as otoacoustic emissions and speech perception in noise, are preserved rather well. The main effect of the defects is a shift in the operation point of the outer hair cells with near intact functioning at high levels. As most test results reflect those found in middle-ear conductive loss in both families, the sensorineural hearing impairment may be characterized as a cochlear conductive hearing impairment. PMID- 17136633 TI - [The ESCRS study on antibiotic prophylaxis for endophthalmitis following cataract surgery]. PMID- 17136634 TI - [Pharmaceutical compounding in ophthalmology: a therapeutic need]. PMID- 17136635 TI - [ROP: unify screening criteria]. PMID- 17136636 TI - [National survey on vitreo-retinal surgery and the management of rhegmatogenous retinal detachments in Spain. Project retina 2]. AB - PURPOSE: To obtain information throughout Spain on the current management of vitreo-retinal surgical diseases, the number of ophthalmologists involved in retinal surgical diseases care, and the urgent management of retinal detachments. METHODS: A 30 item questionnaire was sent by mail, on two separate occasions, to the heads of Ophthalmology Departments of 276 centers in Spain. RESULTS: A total response rate of 48.2% was achieved, however the answers from private centers represented only 9.4% of the total so these were excluded from the analysis. The centers most likely to respond were those in Teaching Hospitals (TH) (52.3%). Of these, 64% of hospitals responded that, in the last year, they performed pars plana vitrectomies (PPV) and 70% performed more than 100 PPVs in the year. In all, 77% of the centers that performed PPVs had also performed them urgently. A total of 75% of hospitals indicated that they had ophthalmologists mainly dedicated to retinal diseases care, with a median of 2 specialists per center, with this representing 23% of their total staff. Retinal surgery was performed by general ophthalmologists in 11.5% of centres. Only 40.3% of hospitals audit their results (37.5% TH). CONCLUSIONS: Despite the existence of a relatively low response rate, as well as some biases and methodological problems, data on the management of retinal surgical disease has been obtained for the first time at a national level. This data will facilitate later studies and must be taken into consideration in improving the planning and adequate management of these diseases in Spain. PMID- 17136637 TI - [IGF-I, VEGF and bFGF as predictive factors for the onset of retinopathy of prematurity (ROP)]. AB - OBJECTIVE: To determine if IGF-I, VEGF and bFGF, present in the serum of premature infants, are independent risk factors of the development of ROP. It was also our objective to design a multivariate model that included these three cytokines as indicator parameters in the ROP screening, in addition to the other parameters already in existence. METHODS: 74 patients were recruited with a birth weight below 1500g or gestational age below 32 weeks. These were classified into those who developed ROP (N = 37) and those without ROP (N = 37). We obtained serum from each infant at the time of the first examination at 4-6 postnatal weeks. These samples were frozen until the time of analysis. The roles of gestational age and birth weight were also evaluated. RESULTS: There were significant differences in the amount of the cytokines IGF-I and VEGF between the groups with or without ROP, but there were no significant differences for bFGF. The differences enabled us to establish a multivariate model including IGF-I and VEGF for the prediction of risk of ROP. CONCLUSIONS: Cytokine serum levels in premature infants can be useful as an indicator in ROP screening, as well as being used to predict the probability of suffering the illness. PMID- 17136638 TI - [Exploration of the anterior segment by optical coherence tomography-3]. AB - PURPOSE: To show the utility of optical coherence tomography (OCT) for studying the anterior segment and to explain its potential advantages as compared to ultrasonic biomicroscopy (UBM). MATERIALS AND METHODS: We have described the findings in 5 patients with different pathologies of the anterior segment, all of whom were studied with UBM and OCT-3 adjusted for the anterior segment. There were 2 cases of an acute attack of closed angle glaucoma, 1 case of rubeosis iridis, 1 case of penetrating ocular trauma and 1 case of a primary stromal iris cyst. RESULTS: OCT was found to be as useful as UBM in detecting angle closure in patients affected by an acute attack of glaucoma. In addition, OCT was more comfortable for the patient and faster than UBM in obtaining images, with the exploration mean time using OCT being less than five minutes while that with UBM was over ten minutes. OCT has also been demonstrated to be a safe and valuable non-contact examination in other iris pathologies such as rubeosis iridis, for checking the permeability of iridotomies and even for studying iris masses such as primary stromal iris cysts. In these entities OCT may reach a higher resolution than ultrasonic biomicroscopy. CONCLUSION: OCT-3, with an adjustable focus, can obtain images from the scleral angle, as well as from other ocular structures like the iris, thus assisting in the diagnosis of numerous pathologies. PMID- 17136639 TI - [Conjunctival inclusion cyst after strabismus surgery by hang-back recession]. AB - CASE REPORT: A 24-year-old woman, with a history of infantile esotropia and DVD operated on in infancy, had strabismus surgery performed by us. Four months later she presented with a cystic lesion that recurred after drainage and medical treatment. Complete excision of the cystic lesion was therefore performed. DISCUSSION: The epithelial cells implanted on the sclera at the time of the most recent surgery may have been the origin of inclusion cyst which developed after the strabismus surgery. This suggests a possible relationship with the scleral suture as the mechanism of cyst formation, independent of the muscle position. Complete excision is the recommended treatment for large cysts. PMID- 17136640 TI - [Acute angle-closure glaucoma resulting from treatment with nebulised bronchodilators]. AB - CASE REPORT: A 78-year-old woman, with an acute exacerbation of chronic obstructive airways disease, was treated with nebulised ipratropium bromide and salbutamol. Twenty hours after beginning this treatment, she developed acute angle-closure glaucoma (AACG) in her left eye which resolved rapidly with appropriate treatment. DISCUSSION: Nebulised ipratropium bromide and salbutamol increases the intraocular pressure and may cause an AACG in susceptible patients (those with a shallow anterior chamber, hypermetropia, or chronic angle-closure glaucoma). Increased vigilance in such patients treated with these bronchodilators may avoid this adverse effect. Ensuring the mask is correctly fitted, using a T-piece or unvented nebuliser and protective eye wear, are some of the many recommendations made to minimize the development of AACG in these patients. PMID- 17136641 TI - [Topical cyclosporine in the treatment of ocular actinic prurigo]. AB - CASE REPORT: A 14-year-old girl from Peru suffered severe limbitis and conjunctivitis. She also presented with clinical skin features diagnosed as actinic prurigo (AP). Her symptoms were successfully controlled with sustained topical therapy of 2% Cyclosporine A. DISCUSSION: AP is an idiopathic photodermatosis that affects mainly the hispanic population of Latin America. There are ocular signs of severe limbitis and conjunctivitis (like atopic keratoconjunctivitis) in 45% of cases. Literature on the subject is very limited and currently topical cyclosporine seems to be the best therapy available. PMID- 17136643 TI - [Naples, April 1909]. PMID- 17136644 TI - [Parametric biomedical imaging--what defines the quality of quantitative radiological approaches?]. AB - Quantitative parametric imaging approaches provide new perspectives for radiological imaging. These include quantitative 2D, 3D, and 4D visualization options along with the parametric depiction of biological tissue properties and tissue function. This allows the interpretation of radiological data from a biochemical, biomechanical, or physiological perspective. Quantification permits the detection of small changes that are not yet visually apparent, thus allowing application in early disease diagnosis and monitoring therapy with enhanced sensitivity. This review outlines the potential of quantitative parametric imaging methods and demonstrates this on the basis of a few exemplary applications. One field of particular interest, the use of these methods for investigational new drug application studies, is presented. Assessment criteria for judging the quality of quantitative imaging approaches are discussed in the context of the potential and the limitations of these methods. While quantitative parametric imaging methods do not replace but rather supplement established visual interpretation methods in radiology, they do open up new perspectives for diagnosis and prognosis and in particular for monitoring disease progression and therapy. PMID- 17136645 TI - Experimental phantom lesion detectability study using a digital breast tomosynthesis prototype system. AB - PURPOSE: To compare the sensitivity of conventional two-dimensional (2D) projection imaging with tomosynthesis with respect to the detectability of mammographic phantom lesions. MATERIALS AND METHODS: Using a breast tomosynthesis prototype based on a commercial FFDM system (Siemens MAMMOMAT Novation), but modified for a wide angle tube motion and equipped with a fast read-out amorphous selenium detector, we acquired standard 2D images and tomosynthesis series of projection views. We used the Wisconsin mammographic random phantom, model RMI 152A. The anode filter combinations Mo/Mo and W/Rh at two different doses were used as typical radiographic techniques. Slice images through the phantom parallel to the detector were reconstructed with a distance of 1 mm employing a filtered back-projection algorithm. The image data sets were read by five radiologists and evaluated with respect to the detectability of the phantom details. RESULTS: For all studied radiographic techniques, the detection rate in the tomosynthesis mode was 100 %, i. e. 75 true positive findings out of 75 possible hits. In contrast, the conventional projection mode yielded a detection rate between 80 and 93 % (corresponding to 60 and 70 detected details) depending on the dose and X-ray spectrum. CONCLUSION: Tomosynthesis has the potential to increase the sensitivity of digital mammography. Overlapping structures from out of-plane tissue can be removed in the tomosynthesis reconstruction process, thereby enhancing the diagnostic accuracy. PMID- 17136646 TI - [Characterization of sonographically detected breast lesions using three dimensional data sets]. AB - PURPOSE: The diagnostic value of 3D ultrasound (US) was evaluated in comparison to 2D US on the basis of a set of pre-defined criteria for breast lesions. The individual criteria were correlated with the histological findings and they were ranked according to their significance for lesion characterization in both 3D US and 2D US. MATERIALS AND METHODS: A total of 92 breast lesions were examined of which 61 were malignant and 31 were benign, as confirmed by histology of core biopsy specimens (Aplio 80, Toshiba, Otawara, Japan). The 2D and 3D data sets were stored digitally. The vascular tree was stored separately using 3D fusion. In addition, a power Doppler (PD) examination was performed. A total of 644 individual images were evaluated by two independent readers. The lesions were evaluated using established criteria (BI-RADS classification). For statistical evaluation, the individual US measurements were classified according to the following two groups: those suggesting malignant lesions and those suggestive of benign lesions. Pearsons's chi-square test was performed for both groups. The 2D and 3D techniques were compared directly using kappa (kappa) statistics. Odds ratios (OR) were determined for the purpose of weighting the individual criteria. The sensitivities and specificities of 2D and 3D US were calculated. RESULTS: The diagnostic value of 3D US was comparable to that of 2D US, with only slight differences observed for specific measurements. Analysis of 2D US data indicated that there was a strong correlation between malignant tumor growth and two specific 2D criteria: lesion margin and effect of the lesion on adjacent breast tissue (r (2) = 0.632, p < 0.001). Analysis of 3D images indicated that the significant measurements for tumor malignancy were shape and lesion margin in sagittal, transverse (r (2) = 0741, p < 0.001), and frontal views (r (2) = 0.592, p < 0.001) as well as retraction and lesion demarcation in the frontal views (r (2) = 0.649, p < 0.001). The comparison of 2D and 3D yielded primarily moderate (kappa = 0.412) to good (kappa = 0.789) agreement between the individual criteria. Crucial criteria for lesion characterization were the effect of the lesion on adjacent breast tissue (OR 47.8 [16.0-143.3]) in 2D US and the lesion margin (OR 36.4 [15.3-86.6]) and the 3 additional criteria of the frontal plane (OR 21.9-32.9) in 3D US. The two US techniques had comparable sensitivities (2D/3D 95-97 %/92-95 %) and specificities (2D/3D 54-58 %/61-67 %). The reconstructed 3D view of the vascular tree was subjectively considered to be helpful by both readers. A higher degree of vascularization was observed with 3D US. CONCLUSIONS: The diagnostic role of 3D ultrasound in the diagnostic evaluation of breast cancer is comparable to that of 2D US. Our findings illustrate the significance of the evaluation of frontal views in lesion characterization. PMID- 17136647 TI - [Trabecular bone mineral density measured by quantitative CT of the lumbar spine in children and adolescents: reference values and peak bone mass]. AB - PURPOSE: The aim of this study was to assess bone density values in the trabecular substance of the lumbar vertebral column in children and young adults in Germany from infancy to the age of peak bone mass. MATERIALS AND METHODS: We performed quantitative computed tomography (QCT) on the first lumbar vertebra in 28 children and adolescents without diseases that may influence bone metabolism (15 boys, 13 girls, mean ages 11 and 8 years, respectively). We also measured 17 healthy young adults (9 men, 8 women, mean ages 20 and 21 years). We used a Somatom Balance Scanner (Siemens, Erlangen) and the Siemens Osteo software. Scan parameters: Slice thickness 1 cm, 80 kV, 81 or 114 mAs. We measured the trabecular bone density and the area and height of the vertebra and calculated the volume and content of calcium hydroxyapatite (Ca-HA) in the trabecular substance of the first lumbar vertebra. RESULTS: Prepubertal boys had a mean bone density of 148.5 (median [med] 150.1, standard deviation [SD] 15.4) mg/Ca-HA per ml bone, and prepubertal girls had a mean density of 149.5 (med 150.8, SD 23.5) mg/ml. We did not observe a difference between prepubertal boys and girls. After puberty there was a significant difference (p < 0.001) between males and females: Mean density (male) 158.0, med 162.5, SD 24.0 mg/ml, mean density (female) 191.2, med 191.3, SD 17.7 mg/ml. The Ca-HA content in the trabecular bone of the first lumbar vertebra was 1.1 (med 1.1, SD 0.5) g for prepubertal boys and 1.1 (0.9, 0.4) g for prepubertal girls. For post-pubertal males, the mean Ca-HA content was 3.5 g, med 3.5, SD 0.5 g, and for post-pubertal females, the mean content was 2.8, med 2.7, SD 0.4 g. CONCLUSION: The normal trabecular bone mineral density is 150 mg/ml with a standard deviation of 20 mg/ml independent of age or gender until the beginning of puberty. Peak bone mass (bone mineral content) in the trabecular substance of the lumbar vertebral column is higher in males than in females, and peak bone density is higher in young female adults than in young male adults. PMID- 17136648 TI - [Radiofrequency ablation of malignant liver tumors: use of a volumetric necrosis tumor ratio for local control]. AB - PURPOSE: Sufficient safety margins are essential for preventing local tumor recurrence after radiofrequency ablation RFA of malignant liver tumors. The aim was to determine the initial tumor volume, ablation necrosis volume, and the necrosis-tumor quotient in order to compare these parameters with the rate of local control during follow-up. MATERIALS AND METHODS: 35 patients with 53 tumor nodules (29 colorectal metastases and 24 HCC nodules) were enrolled. RFA procedures were performed under CT guidance with intravenous conscious sedation. Tumor volumes were measured based on CT data sets and the necrosis volume was assessed using the sum-of-area method. A volumetric necrosis/tumor quotient (NTQ) was calculated for all lesions. Follow-up examinations were performed after 3, 6, and 12 months and then on a yearly basis to identify local recurrent tumors. RESULTS: The CRC metastases and HCC nodules had a median tumor volume of 8.3 ml and 7.4 ml, respectively. The mean ablation volumes were 37.6 ml and 29.5 ml, respectively. This resulted in a median NTQ of 3.9 for metastases and 3.4 for HCC. The follow-up (mean time 18 months) revealed local tumor recurrence in 16 of 29 (55 %) metastases and 10 of 24 (42 %) HCC nodules. In lesions with local recurrence, the initial tumor volume was significantly greater and the NTQ was significantly smaller. A threshold value of 3.4 for NTQ has the highest predictive value for local tumor recurrence. CONCLUSION: The volumetric necrosis/tumor quotient NTQ makes it possible to predict the local outcome and can be used for the planning of additional therapy. PMID- 17136649 TI - [Success and complication rate of CT-guided marking of pulmonary nodules with coil wires for video-assisted thoracoscopic surgery (VATS)]. AB - PURPOSE: To assess the success and complication rate of the CT-guided marking of pulmonary nodules for video-assisted thoracoscopic surgery (VATS). MATERIALS AND METHODS: Pulmonary nodules (mean diameter 9 +/- 5 mm, mean pleural distance 7 +/- 5 mm) were marked with a coil wire in 30 patients (20 males, mean age 57.6 +/- 15.5 years, 22 patients with a history of malignancy). The intended coil-nodule distance was < or = 10 mm. RESULTS: 81 % of nodules were not visible by thoracoscopy. The technical success rate of CT-guided marking was 86.7 %. The projected nodule-coil distance was achieved in 90 % of cases. The procedure had to be changed from thoracoscopy to thoracotomy in 4 patients due to coil wire marking problems: 2 x coil displacement, 1 x coil-nodule distance > 10 mm, unfavorable direction of wire. Histology was determined in all patients (70 % malignant, 30 % benign). Complications requiring therapy were not observed. CONCLUSIONS: The CT-guided marking of pulmonary nodules is a precondition for VATS if the nodule does not involve the visceral pleura in the majority of cases. The success rate is high with a low complication rate. PMID- 17136650 TI - [Magnetically based enhancement of nanoparticle uptake in tumor cells: combination of magnetically induced cell labeling and magnetic heating]. AB - PURPOSE: Magnetic nanoparticles (MNP) are known to be versatile tools in diagnostic and interventional radiology. The goal of the present study was to assess whether MNP can be selectively accumulated on human adenocarcinoma cells in vitro using an external magnetic field (magnetically induced cell labeling) and whether these labeled tumor cells can then be destroyed after being exposed to an alternating magnetic field (magnetically induced heating). In this context, a long-term goal is to combine these two developing methods to achieve an additive effect in tumor therapy. MATERIALS AND METHODS: BT-474 cells were incubated until confluence. Magnetic nanoparticles (0.32 mg Fe/ml culture medium) were then added and the flask was exposed to an external magnetic field gradient (magnetically induced cell labeling, 56 or 83 mT magnets) for 24 hours in order to label the tumor cells with nanoparticles. Cells without both MNP and magnetic labeling as well as cells with MNP incubation but without magnetic labeling served as controls. After MNP incubation, the magnetically labeled cells (5 x 10 (7) cells/ml) were exposed to an alternating magnetic field for 5.45 minutes (frequency 400 kHz, amplitude 24.6 kA/m). The combination effect of both magnetic labeling and magnetic heating was assessed by determining the temperature increase. The amount of MNP accumulated within the cells was determined by measuring the iron content via atomic absorption spectrometry. For statistical analysis mean values and standard deviations of temperature increases and iron contents were calculated and the differences were analyzed using the Student's t test. RESULTS: A significant temperature increase (p < 0.01) during magnetic heating of 41.76 +/- 4.60 K was detected after magnetic labeling of the cells (5 x 10 (7) cells/ml, 83 mT) incubated with MNP. In comparison, the cells incubated with MNP but without magnetic labeling revealed a temperature increase of 32.03 +/- 3.33 K, naked cells of only 2.69 +/- 0.34 K. CONCLUSION: The results demonstrated the magnetically based enhancement of cellular uptake of nanoparticles by tumor cells, resulting in the intensification of the generated temperature increase during magnetic heating. Consequently, magnetic nanoparticles are shown to be valuable tools for the combination of magnetically based therapy modalities. PMID- 17136651 TI - [Choriocarcinoma]. PMID- 17136652 TI - [Cryptogenic organizing pneumonia after metal fume inhalation]. PMID- 17136653 TI - [Isolated tracheobronchial amyloidosis: a rare cause of a hilar space-occupying lesion]. PMID- 17136655 TI - [Transient elastography for diagnosing liver cirrhosis]. AB - BACKGROUND AND OBJECTIVE: Detection of liver cirrhosis has numerous implications because of the potential sequelae of cirrhosis. Transient elastography (Fibroscan), was evaluated as a novel, non-invasive means of assessing cirrhosis by measuring liver stiffness. METHODS: 147 consecutive patients with different forms of liver disease and histologically determined stages of liver fibrosis were prospectively studied by transient elastography. 48 patients had liver cirrhosis. RESULTS: The number of transient elastographic measurements per patient was 12+/-4 (range 6 - 30). Valid elastography measurements were available for 135 out of 147 patients (92 %). The results of transient elastography correlated positively with the histological score of liver fibrosis (r = 0.8; 95 % CI: 0.72 - 0.85; p < 0.001). Areas under the receiver operating characteristic curve (AUROC) were 0.91 for > or = F3 fibrosis (95 % CI: 0.85 - 0.96) and 0.94 for cirrhosis (95 % CI: 0.90 - 0.98). Using a cut-off value of 13 kPa for detection of liver cirrhosis a sensitivity of 90 %, a specificity of 82 %, a positive predictive value of 71 % and a negative predictive value of 95 % were obtained. CONCLUSIONS: Measuring liver stiffness by transient elastography proved to be an easy method to assess liver cirrhosis. In combination with clinical signs, ultrasound and biochemical markers noninvasive diagnosis of liver cirrhosis will be further improved. PMID- 17136656 TI - ["Ghost peak" in gas chromatography in a delirious woman with severe metabolic acidosis. Intoxication with an unknown substance?]. AB - HISTORY AND CLINICAL FINDINGS: A 31-year-old female with known type 1 diabetes mellitus was referred because of symptomatic hyperglycemia. On admission she was delirious and impressed with marked Kussmaul breathing. All other vital signs were normal. INVESTIGATIONS: Blood serum glucose concentration was 26.4 mmol/l. Arterial blood gas analysis revealed massive metabolic acidosis (pH 6.80) with an elevated anion gap (21 mmol/l) and a marginally increased osmolar gap (21,5 mOsm/l). TREATMENT AND COURSE: Despite normalization of the serum glucose and acidemia after administration of normal saline, insulin and bicarbonate, the delirium persisted, and the possibility of an additional intoxication had to be considered. Serum headspace analysis for intoxication with solvents (gas chromatography) finally detected a "ghost peak", which could not be assigned to any established substance. The same peak was, however, found in a healthy subject's serum and was found to be a "toluene peak". Toluene is contained as "contaminator" in gels in blood collection tubes. The patient gradually regained consciousness and "merely" suffered from diabetic ketoacidosis associated with cocaine use. CONCLUSION: The differential diagnosis of high anion gap metabolic acidosis includes among other reasons intoxications with different kinds of solvents. When looking for solvents in the serum when poisoning is suspected (headspace analysis), only blood collection tubes without gel (EDTA plasma) should be used, because all gels contain solvents (in this case toluene). PMID- 17136657 TI - [Superior vena cava syndrome: Catheter association as a rare cause]. AB - HISTORY AND FINDINGS: A 78 year-old-woman was admitted with a swollen face, edema of the lower eyelids and dyspnea. The past medical history revealed an ovarian carcinoma treated with polychemotherapy. Half a year before the patient had been investigated for similar clinical symptoms but no underlying cause had been detected. The histology of an enlarged axillary lymph node did not show a malignancy. The symptoms persisted after angioedema-inducing drugs had been discontinued. INVESTIGATIONS: Initial CT scan, magnetic-resonance tomography as well as positron emission tomography failed to explain the clinical findings. Also, testing of serological, immunological and endocrinological tests as well as the differential blood count did not reveal a likely cause of the clinical symptoms. However, 4 weeks later, a repeat CT scan showed a stenosis of the superior vena cava (SVC) establishing the diagnosis of an SVC syndrome. TREATMENT AND COURSE: The port catheter tipp was localized horizontal to the vena cava superior and was touching the vein wall. Removal of the catheter and subsequent balloon dilatation of the stenosis immediately lead to a reduction of the eyelid swelling. CONCLUSIONS: Hence, in a case of an SVC-syndrome, complete stenosis caused by an implanted venous access system should be considered though it is rare. PMID- 17136659 TI - [Neuromuscular complications of sepsis: "Critical illness" polyneuropathy and myopathy]. PMID- 17136660 TI - [Immunointervention in newly diagnosed type 1 diabetes]. PMID- 17136661 TI - [Water filter for prevention of nosocomial legionellosis?]. PMID- 17136662 TI - [Epilepsy: which therapy in acute care?]. PMID- 17136663 TI - [Defibrillation done by layman in Germany: particular conditions?]. PMID- 17136664 TI - [Interdisciplinary guidelines for the diagnosis and therapy of extracerebral amyloidosis]. PMID- 17136667 TI - [Diagnostic and therapeutic management of patients receiving antithrombotic drugs: what to heed?]. PMID- 17136670 TI - Reconstruction of large orbital exenteration defects after resection of periorbital tumors of advanced stage. AB - Orbital exenteration is a devastating procedure because of the social impact and psychological stress put upon the patient. Besides cancer ablative surgery, reconstruction after removal of the tumor constitutes a major problem and the final aesthetic result is quite important. Both obliteration of the orbital cavity and continuation of the epithelial lining are required. Free rectus abdominis muscle and musculocutaneous flaps are versatile flaps which both enable filling the cavity and reconstituting the skin defect with a cutaneous portion or with a skin graft. Both free rectus abdominis muscle and musculocutaneous flaps were used for reconstruction of orbital exenteration defects in 19 patients. All suffered partial maxillectomy as well. The flaps provided satisfactory aesthetic results in all patients. PMID- 17136671 TI - Free and island flap transfers for the treatment of motor vehicle injuries. AB - Thirteen patients (10 males and 3 females; age range at surgery: 2 to 41 years, average: 13.5 years) with motor vehicle injuries of the foot (n = 10) and leg (n = 3) were treated using free and island flaps. Donor flaps consisted of a scapular flap in six cases, a peroneal flap in four, a latissimus dorsi musculocutaneous flap in one, gracillis muscle flap in one, and gastrocnemius muscle flap in one case. Free grafts were transferred in 10 patients and pedicle grafts in three. All flaps survived completely. Moderate scarring was noted in two patients who underwent scapular flap transfer, and growth disturbances occurred in two patients with metatarsal fracture accompanying epiphyseal plate injury. Functionally, no patients experienced any disturbance, and all reconstructed sites were cosmetically satisfactory. Early flap coverage of soft tissue defects due to motor vehicle injury was shown to be useful from the point of view of both cosmetic and functional results. PMID- 17136672 TI - Free flap choice for soft tissue reconstruction of the severely damaged upper extremity. AB - Reconstruction of complex wounds of the hand associated with severe bone, tendon, nerve and soft-tissue injuries has been a major problem in hand surgery. Early definitive soft-tissue coverage of this kind of extensive wound with well vascularized tissue is one of the most important stages of reconstruction for salvage of the extremity and restoration of function. Although multiple free flap donor sites have been described for complex upper extremity wounds, the authors think that anterolateral thigh (ALT) and lateral arm (LA) flaps are good choices for soft-tissue reconstruction in the upper extremity because of their reconstructive characteristics. These flaps can be used as flow-through and also sensate flaps. There is no need for position change intraoperatively and two teams may work simultaneously. Donor sites can be hidden and there is no required sacrifice of major artery or muscle. PMID- 17136673 TI - Limited myocardial muscle necrosis model allowing for evaluation of angiogenic treatment modalities. AB - The currently accepted model for creating infarcted cardiac tissue in a rat model involves ligation of the left anterior descending artery (LAD), either proximally or at the bifurcation level. This procedure requires significant technical expertise and, even in skilled hands, commonly results in a 30% to 60% animal mortality. The authors propose a new model for creating a limited area of myocardial muscle necrosis that can be effectively studied. It involves a distal electrocautery occlusion of the LAD terminal branches and coagulation of the surrounding muscle. The model is consistently reproducible and decreases the morbidity of the study animals. It provides a cardiac muscle necrosis model not dependent on survival, while allowing study of the post injured state of the muscle and surrounding scar. This allows researchers to evaluate neovascularization and healing of the scar and peri-necrotic muscle, to assess improving blood flow with treatment by techniques designed to improve and stimulate angiogenesis, and to measure the outcome of stem-cell transplants for potential clinical use. PMID- 17136674 TI - Posterior-wall-first continuous suturing combined with conventional interrupted suturing for microvascular anastomosis. AB - Vascular anastomosis with conventional interrupted suturing is often difficult to perform when the vascular clamp is not reversed, because of a narrow operative field or a short vascular pedicle. A posterior-wall-first continuous suture technique combined with the standard interrupted suture technique is one method of solving this problem. The authors conducted a comparative study of posterior wall-first continuous suturing combined with standard interrupted suturing and conventional interrupted suturing in rat vessels. There was no statistically significant difference in patency rates or suturing times. Electron microscopy demonstrated no significant difference in recoverability of the intima. The posterior-wall-first continuous suture technique combined with the standard interrupted suture technique is a useful alternative to the conventional interrupted suture technique. PMID- 17136675 TI - New microvenous anastomosis model for microsurgical training: external jugular vein. AB - In microsurgical training, the femoral vein is used frequently for a microvenous anastomosis model. But the femoral vein in the rat does not completely simulate the human vein because of its thin wall, fragility, and tendency to collapse. These anatomic characteristics cause some difficulty in carrying out anastomoses in microsurgery training particularly for beginners. The authors propose the external jugular vein of the rat for microsurgical training in microvenous anastomoses. In 10 Wistar rats, the anatomy of the external jugular vein was studied by dissection and histology. Anatomic dissections demonstrate that the external jugular vein has an average diameter of 1.9 mm (range: 1.6 to 2.1 mm) without tendency to collapse. The vein is easily dissected without any accompanying anatomic structure for an average segment of 45 mm, allowing effortless approximator clamp placement. Comparison of its cross section with that of the femoral vein and other previously described models by light microscopy and scanning electron microscopy reveals a larger diameter and much thicker vessel wall with a prominent tunica media and adventitia. Based on the anatomic findings in 20 rats, the external jugular vein was anastomosed with end to-end standard microsurgical technique using 8-0 (n = 10) and 10-0 (n = 10) nylon sutures. Results indicate a 100 percent patency rate immediately after the anastomosis for the two subgroups and 100 percent and 90 percent patency rates 1 week after the procedure for the 10-0 and 8-0 nylon suture groups, respectively. This model presents some advantages: the vein is easily dissected with the naked eye without using the operating microscope because it is the largest vein among the superficially located veins in the rat, and has a thick vessel wall without tendency to collapse. The operative area allows for training inbilateral microsurgical anastomoses using a single skin incision and is safe from autocannibalization. The model simulates clinical microvenous anastomosis better because of its similarities to human large diameter flap veins. PMID- 17136676 TI - End-to-end microvascular anastomosis in the rat carotid artery using continuous horizontal mattress sutures. AB - This paper reports a continuous horizontal mattress suture technique with advantages such as decreased time for anastomosis, minimized anastomotic leakage, eversion around the vessel edges, and other advantages which the continuous anastomosis technique has. This technique was compared with the classical interrupted and classical continuous suture techniques on a total of 59 Sprague Dawley rat common carotid arteries: Group 1 (n = 19), interrupted suture technique; Group 2 (n = 20), standard continuous technique, and Group 3 (n = 20), continuous horizontal mattress technique. Early (30 min) and late (21 days) patency rates, anastomosis time, leakage on clamp release, oozing duration, additional sutures needed, and total number of sutures placed were statistically compared between groups. Specimens were taken at the 21st day randomly, and light microscopy (LM), scanning electron microscopy (SEM), and angiographic studies were performed. Results revealed that the continuous mattress suture technique has the advantages of providing a water-tight anastomosis with less suture materials in a shorter time, and minimal intraluminal suture material which can incite thrombosis. On the other hand, a tendency to anastomotic stricture was found to be the sole disadvantage of this technique. PMID- 17136677 TI - Proinflammatory cytokines gene expression in skin flaps with arterial and venous ischemia in rats. AB - Infiltration of inflammatory cells is the crucial element in ischemia-reperfusion injury of the microsurgical flap. Cytokines are a large functional group of polypeptide regulatory molecules that influence the activity of various cell types through autocrine and paracrine mechanisms. In this study, expression of selected proinflammatory cytokines was examined in skin flaps with arterial and venous ischemia in the rat model. Fifty-four Sprague-Dawley rats were used in the study. The ischemia of each flap was induced by clamping its vascular pedicle for 6 hr. The flap was then replaced and allowed to reperfuse. All flaps were biopsied immediately post-event, and at 3, 6 and 18 hr after reperfusion. Expression of tumor necrosis factor (TNF-alpha), interleukin (IL-1beta), and monocyte chemoattractant protein-1 (MCP-1) mRNA was determined by RT-PCR in each case. The same number of skin flaps without ischemia was used for baseline gene expression. Results showed that the TNF-alpha expression was significantly up regulated in the flaps with arterial ischemia at 6 hr after reperfusion. In the flaps with venous ischemia, MCP-1 expression was increased with its peak expression at 3 hr after reperfusion. IL-1beta expression was increased threefold in the flaps subjected to venous ischemia and following reperfusion in 3 hr, but the peak expression in the flap with arterial ischemia was observed at 18 hr after reperfusion. This study delineated the changes in expression of these proinflammatory cytokines in flaps with arterial and venous ischemia reperfusion injury, and showed that cytokine expression was different in the arterial and venous injuries. PMID- 17136678 TI - Effects of intrathecal administration of FK506 after sciatic nerve crush injury. AB - Although various administration routes of FK506 have been published, intrathecal administration of FK506 has not previously been reported in the literature. A daily dose of 0.05 mg/kg of FK506 was given (a small dose compared with those reported in the available literature). The authors used this small dose to obtain lower immunosuppression and neurotoxicity, and a higher axonal regeneration rate. A total number of 40 female Wistar rats were used and randomly divided into four groups: control, sham, FK506-treated, and vehicle-treated. Sciatic nerve regeneration was evaluated by walking track analysis, an electrostimulation test, and light microscopic evaluation. There was a statistically significant difference ( P < 0.05) between FK506-treated and vehicle-treated groups at the end of 6 weeks according to both the walking track analysis and the electrostimulation test. Comparing the stimulus thresholds of the sham and FK506 treated group, no significant difference ( P > 0.05) was observed. Evaluation of the data revealed that FK506 had a beneficial effect on sciatic nerve regeneration. PMID- 17136679 TI - Bone nanostructure near titanium and porous tantalum implants studied by scanning small angle x-ray scattering. AB - Bone sections including either titanium or porous tantalum implant devices used for interbody spinal fusion were investigated with position-resolved small angle X-ray scattering (sSAXS). The samples were obtained from six-month-old pigs that had undergone surgery three months prior to sacrifice. The aim of the study was to explore the possibility of using sSAXS to obtain information about thickness, orientation and shape/arrangement of the mineral crystals in bone near the implant surfaces. Detailed sSAXS scans were carried out in two different regions of bone adjacent to the implant in each of the implant samples. In the implant vicinity the mineral crystals tended to be aligned with the surface of the implants. The mean crystal thickness was between 2.1 and 3.0 nm. The mineral crystal thickness increased linearly with distance from the implant in both regions of the porous tantalum implant and in one of the regions in the titanium sample. In the second region of the titanium sample the thickest mineral crystals were found close to the implant surface. The observed differences in mineral thickness with distance from the implant surfaces might be explained by differences in mechanical load induced by the implant material and the geometrical design of the implant. The study shows that sSAXS is a powerful tool to characterize the nanostructure of bone near implant surfaces. PMID- 17136680 TI - The structure and function of cartilage proteoglycans. AB - Cartilage contains a variety of proteoglycans that are essential for its normal function. These include aggrecan, decorin, biglycan, fibromodulin and lumican. Each proteoglycan serves several functions that are determined by both its core protein and its glycosaminoglycan chains. This review discusses the structure/function relationships of the cartilage proteoglycans, and the manner in which perturbations in proteoglycan structure or abundance can adversely affect tissue function. PMID- 17136681 TI - Clinical databases in nephrology: research and clinical practice goals and challenges. AB - In clinical practice we formulate direct questions related to patient management that should be answered on the basis of results of valid studies. Bias problems are dealt with using different approaches in observational studies and in clinical trials. The clinical trial is the standard for assessing the efficacy of treatments while the efficiency of treatments at community level is better captured by observational studies. Electronic medical records have now emerged as a precious, matchless data source for clinical audits. The value of an audit cannot be taken for granted and should be tested in the specific health care setting where it is applied. Electronic medical records and high quality clinical databases offer a great opportunity for performing observational studies and for reducing the cost of clinical trials. Research on audit and feedback functionality is a new, useful and stimulating research area which may be of great interest to nephrologists. PMID- 17136682 TI - Acute kidney dysfunction secondary to the abdominal compartment syndrome. AB - The abdominal compartment syndrome (ACS) occurs most commonly in the setting of major trauma and complex abdominal surgical procedures. The syndrome reflects the adverse physiological consequences of an acute increase in intra-abdominal pressure (generally >18 mm Hg). The effects of increased abdominal pressures on the kidney were initially described in 1876 and include impairment of renal blood flow and glomerular filtration resulting in oliguria or anuria and acute kidney dysfunction. These effects are magnified by the concomitant effects of increased intra-abdominal pressure to impair venous return and cardiac output. Patients with intra-abdominal hypertension (IAH) can be easily detected using simple methodology. If employed early, abdominal decompression to lower IAH is associated with restoration of organ function and avoidance of the ACS. However, the overall mortality associated with this syndrome remains high. In postsurgical, trauma patients, or those at risk, ACS should be considered as a potential etiology for acute kidney dysfunction and intra-abdominal pressures should be measured, monitored and when necessary intervened upon in order to attempt to improve organ dysfunction. PMID- 17136683 TI - Vitamin D retains an important role in the pathogenesis and management of secondary hyperparathyroidism in chronic renal failure. AB - Chronic kidney disease (CKD) causes alterations in mineral metabolism inducing the development of secondary hyperparathyroidism (HPT) and renal osteodystrophy. Recently, it has been suggested that these alterations play an important role in determining extraskeletal calcification and thus cardiovascular morbidity and mortality among CKD patients. An impaired 1 alfa -hydroxylation of 25 hydroxycholecalciferol (25(OH)D3) to 1,25-dihydroxycholecalciferol (1,25(OH)2 D3) with decreased circulating 1,25(OH)2 D3 levels is commonly observed in patients with creatinine clearance below 70 ml/min. The reduction in 1,25(OH)2 D3 production triggers the up-regulation of parathyroid hormone (PTH) synthesis, through a decreased suppression on PTH gene transcription and a decreased intestinal calcium absorption. A reduced expression of vitamin D receptor (VDR) and a less efficient binding of the complex 1,25(OH)2 D3 -VDR to specific DNA segments account for the resistance to 1,25(OH)2 D3 in target cells. Thus, absolute and relative 1,25(OH)2 D3 deficiency is one of the causes of secondary HPT in patients with CKD, together with phosphate retention and skeletal resistance to PTH. Consistently with these pathophysiological mechanisms, the therapeutic use of 1,25(OH)2 D3 still represents a milestone for the treatment of secondary HPT and renal osteodystrophy, even though hypercalcemia and hyperphosphatemia are common adverse events and may increase the risk of cardiovascular calcifications. To reduce the impact of such adverse effects while retaining anti-PTH activity, 1,25(OH)2 D3 analogues with lower calcemic effects have been synthesized and are now available for clinical use. PMID- 17136684 TI - New immunosuppressive drugs for prevention and treatment of rejection in renal transplant. AB - Rejection prevention and control have been pivotal aims of medical research since the time that renal transplant was first developed. While acute rejection rates have generally fallen to 10%-15% at 1 year, chronic rejection and chronic allograft nephropathy (CAN) still erode long-term transplant potential. Furthermore, drug tolerability and their 'secondary' effects other than immunosuppression have become as important as their effectiveness in a recipient population that is increasingly old and burdened by cardiovascular morbidity. We review the state of the art of new immunosuppressive drugs: some of them have already entered clinical practice (e.g., mycophenolate mofetil, anti-CD25 monoclonal antibodies and sirolimus [SRL]); others are at different stages of the pre-registration phase (everolimus, mycophenolic acid, FTY 720, leflunomide-FK 778 and Campath-1H), and many others show promising potential for the future (rituximab, monoclonal antibodies against costimulatory molecules). Especially, new drugs with antiproliferative and antifibrogenic effects (SRL, everolimus, leflunomide and mycophenolic acid) could to be decisive in still challenging areas such as CAN and cardiovascular and neoplastic transplant complications. The key to solving these open issues probably lies in the possibility of relying on a wide repertoire of drugs with different profiles, to be exploited in several associations, and in our capacity to integrate new and old immunosuppressive drugs. PMID- 17136685 TI - The role of the Italian Directory of Nephrology Units. PMID- 17136686 TI - Long-term organ protection by doxazosin and/or quinapril as antihypertensive therapy. AB - BACKGROUND: Even with optimal blood pressure control, organ protection may also depend on the selected therapeutic regime. Angiotensin-converting enzyme inhibitors have been shown to provide excellent organ protection in hypertension, and may show dose-dependent protective effects. Adrenergic alpha blockers have been associated with an increased rate of heart failure in the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT) and Vasodilator-Heart Failure Trial (V-HeFT). This has been related to a proapoptotic effect of this drug in cardiomyocytes. Our purpose is to compare the heart and renal protection of a high quinapril dose, with a combined low quinapril dose plus doxazosin, in an animal model of chronic hypertension. METHODS: Uninephrectomized spontaneously hypertensive 12-week-old rats were treated for 36 weeks with either quinapril or a combination of doxazosin plus a low quinapril dose. Tight blood pressure control was achieved with both treatments. Renal and cardiac protection was assessed by different parameters, and cardiac apoptosis was evaluated by active caspase-3, apoptotic protein and heat shock protein levels. Untreated hypertensive and normotensive rats were included as controls. RESULTS: Both treatments showed significant heart and renal protection compared with untreated animals. Both therapeutic regimes showed similar protection in renal and cardiac pathology, coronary media fibrosis, myocardial apoptosis and cardiac index. Proteinuria and left ventricular hypertrophy regression were significantly lower in the quinapril group compared with the combined treatment group. CONCLUSIONS: Blood pressure control with a high quinapril dose provided higher organ protection than a combined therapy with a lower quinapril dose. This effect was not due to a deleterious effect of doxazosin. PMID- 17136687 TI - Uremic serum induces proatherogenic changes in human endothelial cells. AB - BACKGROUND: Cardiovascular complications are the main cause of death in uremic patients. Uremic angiopathy has been regarded as an accelerated form of atherosclerosis. However the mechanism leading to vessel wall injury is still unknown. We hypothesized that uremic serum affects endothelium inducing a proatherogenic state. METHODS: We studied the effects of uremic serum on human endothelial cells (HECs). Cell proliferation and adhesion of mononuclear cells to HEC monolayers were evaluated by cell counting, apoptosis and collagen production by ELISA, and nitric oxide (NO) by measuring the concentration of nitrite/nitrate in the cell supernatant. (alfa2)IV collagen, tissue inhibitor of metalloproteases 1 (TIMP-1) and transforming growth factor-beta (TGF-beta) mRNA levels were measured by reverse transcriptase polymerase chain reaction (RT-PCR). In some experiments cells were preincubated with anti-receptor for advanced glycation end product (anti-RAGE) blocking antibodies. RESULTS: Uremic serum did not modify HEC proliferation but induced apoptosis after 72 hours of incubation. Adhesion of mononuclear cells to HEC monolayers was significantly increased by uremic serum. In addition, uremic serum increased (alfa2)IV collagen, TIMP-1 and TGF-beta mRNA levels. There was no increase in nitric oxide concentration in ure-mic serum treated endothelial cells, and the expression of TGF-beta was neither modified by L-NAME nor by anti-RAGE antibodies. CONCLUSIONS: Our results indicate that uremic serum affects HEC inducing a proatherogenic state that may be responsible for the accelerated atherosclerosis of uremic patients. Apparently uremic serum effect is not mediated by NO or by AGEs. PMID- 17136688 TI - False positive dipstick for urinary blood in childhood. AB - BACKGROUND: In view of the obvious practical advantages, the most common test for hematuria is currently a reagent strip. METHODS: A standardized microscopic examination of the sediment was performed in 20 asymptomatic children referred for evaluation of chronic isolated microhematuria detected by means of a reagent strip. RESULTS: In 6 of the 20 children the microscopic examination failed to confirm the result of the dipstick test. CONCLUSIONS: Confirmation for the presence of hematuria by microscopy is the most important step in children with a positive dipstick for urinary blood. PMID- 17136689 TI - Prevalence, severity and predictors of HOMA-estimated insulin resistance in diabetic and nondiabetic patients with end-stage renal disease. AB - BACKGROUND: Prevalence of insulin resistance (IR) is increased in type 2 diabetes and in end-stage renal disease (ESRD). IR is associated with advanced atherosclerosis and is an independent predictor for cardiovascular disease in diabetes and ESRD patients. We investigated prevalence, severity, predictors and relation to vascular diseases by the homeostasis model assessment (HOMA-IR) in diabetic and nondiabetic ESRD patients. METHODS: ESRD patients with type 2 diabetes (n = 27) and nondiabetic ESRD patients (n = 35) were included in the study. IR was assessed with the HOMA-IR using fasting glucose and insulin levels. Additionally, serum levels of C-peptide, HbA1c, triglycerides, cholesterol and C reactive protein and blood pressure were assessed. RESULTS: Median HOMA-IR was significantly higher in the diabetic ESRD patients than in the nondiabetic ESRD patients (6.3 [range 0.7-61.7] vs. 2.4 [range 0.3-5.7]; p < 0.001). Systolic blood pressure and triglycerides were significantly higher in patients with higher HOMA-IR, whereas HDL cholesterol was significantly lower in those patients. Only nondiabetic patients with increased HOMA-IR had significantly higher C-peptide levels than those with lower HOMA-IR (14.9 + 5.7 vs. 9.0 + 4.3, p = 0.004). Vascular disease prevalence was significantly higher in diabetic patients with higher HOMA-IR than in those with lower HOMA-IR. CONCLUSIONS: Prevalence and severity of HOMA-IR was greater in diabetic ESRD patients than in those without diabetes. In diabetic patients low HDL cholesterol was the only predictor for higher HOMA-IR, whereas in nondiabetic patients a high C-peptide level was the only predictor for higher HOMA-IR. The prevalence of vascular diseases is associated with higher HOMA-IR in ESRD patients. PMID- 17136690 TI - Oxidative stress, sister chromatid exchanges and apoptosis in the pathogenesis of lymphocytopenia in ESRD patients. AB - BACKGROUND: In end-stage renal disease (ESRD) patients on hemodialysis (HD) there may be a link between oxidative stress, genomic damage and the tendency of peripheral lymphocytes to die by apoptosis. Our aim was to verify this hypothesis, and to ascertain whether the link, if present, could explain lymphopenia in uremic patients. METHODS: The series investigated comprised 55 participants: 30 HD patients on regular maintenance acetate-free bio-filtration (AFB) and 25 age-matched healthy volunteers. One blood sample was drawn from the cubital vein of each participant. In HD patients, samples were drawn 3 times: predialytic, postdialytic and interdialytic (24 hours after the end of the session). Thiobarbituric acid reactants (TBARs), sister chromatid exchange (SCE) rate, high frequency cells (HFCs), total circulating lymphocytes and the percentage of circulating apoptotic lymphocytes were assayed in all samples. A statistical analysis of the findings was made using multiple and linear regression. RESULTS: In AFB patients, TBAR levels appeared higher than in controls, even at baseline (2.15 +/- 0.5 micromol/L vs. 1.20 +/- 0.4 micromol/L; p < 0.05). The highest peak occurred at the end of the session (3.2 +/- 0.4 micromol/L; p < 0.05 vs. basal), and a prompt return to basal values was observed 24 hours later (2.2 +/- 0.6 micromol/L, p < 0.5 vs. basal). In AFB patients, the per-centages of HFCs (8.63% vs. 3%; p < 0.05), SCE (6 +/- 0.6 vs. 4.65 +/- 2.18; p < 0.04) and apoptotic lymphocytes (3-fold) were greater than in controls, even at baseline, whereas the values for total lymphocytes were lower (1,140 +/- 652 vs. 1,590 +/- 822). After an AFB session the differences between patients and control values appeared greater (HFCs, 16.81%, p < 0.04 vs. basal; SCE, 7.02 +/- 1.2, p < 0.03; apoptotic lymphocytes 3.5-fold greater than control values). Twenty-four hours later, a further increase was observed in the expression of genomic damage (HFCs, 50%, p < 0.05 vs. basal; SCE, 9.82 +/- 2.1, p < 0.03) and the percentage of apoptotic lymphocytes (4.7-fold greater than control values), while the lowest peak occurred for total circulating lymphocyte count (997 +/- 854, p < 0.04). At linear regression, a strong positive correlation was found between HFCs and TBARs at the beginning and at the end of the AFB session(r = 0.7, p < 0.03). With multiple regression analysis, a strong positive correlation was found between TBAR levels at the end of AFB session, HFC rate and apoptotic lymphocytes at 24 hours, with the last as the dependent variable (multiple r = 0.8, TBARs, beta = 0.51, p < 0.04; HFCs, beta = 0.43, p < 0.03). DISCUSSION AND CONCLUSIONS: An AFB session has an immediate impact, causing an increase in TBAR levels, genomic da-mage and lymphocytic apoptosis. Twenty-four hours after the session there was a further expression of genomic damage, and an increase in apoptosis, while the peak for lymphocytes dropped sharply. Our findings indicate that lymphopenia affecting end-stage renal disease (ESRD) patients may be strictly related to genomic damage exerted, at least in part, by TBARs, and to a dysregulation in programmed cell death. PMID- 17136691 TI - Comparison of the diuretic effect of furosemide mixed with human albumin or fresh frozen plasma for patients with hypoalbuminemia in the intensive care unit. AB - BACKGROUND: Diuretics are commonly used in the intensive care unit (ICU) for patients with fluid over-loading. Hypoalbuminemia is a major cause of diuretic resistance. Albumin mixed with furosemide can promote diuresis and sodium excretion in patients with hypoalbuminemia. The purpose of this study is to compare the diuretic effect of furosemide (FU) mixed with human albumin (HA) or fresh frozen plasma (FFP) in ICU patients with hy-poalbuminemia. METHODS: Patients with fluid overloading and hypoalbuminemia who needed diuretic treatment were enrolled and were divided into 2 groups: the first group having clearance of creatinine (CCr) >20 ml/min, and the second group having CCr < or = 20 ml/min. FU (60 mg) mixed with HA (HA group), 60 mg FU mixed with FFP (FFP group) and water (placebo group) were given intravenously to these patients for 60 minutes in random order on the first, third and fifth day. After drug administration, 8-hour urine was collected, and urine amount and urinary sodium excretion were checked. RESULTS: Both the HA group and the FFP group had significantly higher urinary volume and sodium excretion than the placebo group in the patients with CCr >20 ml/min or CCr < or = 20 ml/min (p < 0.01). In the patients with CCr >20 ml/min, there was no difference in the amount of urine excretion and cumulative urinary sodium excretion between the HA group and FFP group. In the patients with CCr < or =ml;20 ml/min, the HA group had a significantly higher urine output and urinary sodium excretion than the FFP group (p < 0.05). CONCLUSIONS: In ICU patients, 60 mg FU mixed with HA or FFP has a similar diuretic effect in patients with CCr >20 ml/min. FFP is an effective alternative choice for improving diuresis for ICU patients with hypoalbuminemia. In patients with CCr < or = 20 ml/min, albumin mixed with 60 mg FU has a superior diuretic effect compared with FFP mixed with FU. PMID- 17136692 TI - Relationship of aortic atherosclerosis to acute renal failure following cardiac surgery. AB - BACKGROUND: The presence of aortic atherosclerosis has been identified as a major risk factor for stroke after coronary artery bypass grafting (CABG). Whether aortic atherosclerosis is similarly related to the risk of acute renal failure (ARF), a common and important complication of CABG, is unknown. METHODS: Rates of postoperative ARF were analyzed using data from 1,117 randomized patients in a multicenter controlled trial comparing standard CABG with CABG plus an experimental aortic catheter. Aortic atherosclerosis was prospectively measured using transesophageal echocardiography. The association of aortic atherosclerosis with postoperative renal failure was analyzed using multivariable logistic regression to adjust for confounding by baseline and intraoperative conditions. RESULTS: Baseline creatinine clearance <40 ml/min and systolic hypertension were strong predictors of postoperative acute renal failure. Neither mild nor moderate aortic atherosclerosis was associated with the development of acute renal failure. Patients with moderate aortic atherosclerosis had a lower risk of acute renal failure (odds ratio = 0.53, p = 0.20) than those with lesser degrees of atherosclerosis. CONCLUSIONS: Our results demonstrate that the presence of significant aortic atherosclerosis does not increase the risk of acute renal failure following CABG, and they suggest that cholesterol embolization from the aorta to the renal circulation is an infrequent cause of acute renal failure after bypass surgery. Strategies to decrease cholesterol embolization from the aorta are unlikely to significantly lower the rate of renal failure following bypass surgery. PMID- 17136693 TI - Acute renal failure in intensive care unit: which factors predict future dialysis dependency? AB - Management of acute renal failure (ARF) in an intensive care unit (ICU) is difficult. The aim of this study was to identify prognostic factors determining ARF outcome in the ICU in terms of dialysis dependency or independency. We included 35 patients who turned out to be dialysis dependent (DD) and 11 patients who turned out to be dialysis independent (DI) after ARF in the ICU, which necessitated renal replacement therapy. In the post-ARF period, acetylsalicylic acid was protective against dialysis dependency (p < 0.05, odds ratio [OR] = 0.078) and dopamine increased the likelihood of dialysis dependency (p = 0.016, OR = 10.6). Multiorgan dysfunction (p = 0.001, OR = 13.6), especially cardiac (p = 0.009) and hepatic failure (p < 0.0001) were determined to increase risk of dialysis dependency. Mean systolic blood pressures during the first 24 hours (p = 0.023) and 24-48 hours (p = or < 0.0001), mean diastolic blood pressures during first the 24-48 hours (p = 0.03) and 48-72 hours of ARF in ICU (p = 0.023) and at discharge (p = 0.03) were significantly lower in the DD group than in the DI group. Mean thrombocyte counts at hospitalization (p = 0.034), during the first 24 hours (p = 0.019) and 24-48 hours of ARF in ICU (p = 0.038) were lower in the DD than DI group. This study demonstrates the very early prognostic factors influencing ARF outcome in terms of dialysis dependency. Early thrombocyte count and systolic blood pressure and follow-up diastolic blood pressure were prognostic factors for ARF outcome. Acetylsalicylic acid seemed to improve renal outcome, whereas dopamine seemed to worsen the disease process. PMID- 17136694 TI - Evidence for optimal hemoglobin targets in chronic kidney disease. AB - Even though anemia occurs frequently in patients with chronic kidney disease and therapeutic options are widely available, the ideal hemoglobin target level is not clearly established. We start from 2 anecdotes and review the evidence in favor of and against higher (>12 g/dL) and normal hemoglobin targets as compared to subnormal or low hemoglobin levels in different subsets of chronic kidney disease (either predialysis or dialysis). Current clinical trials and their systematic reviews have found that higher hemoglobin levels (>12 g/dL) do not significantly impact on patient-level cardiovascular end points including cardiovascular and all-cause mortality. Patients feel better, and enhanced quality of life parameters have been identified in most short-term studies when higher hemoglobin levels are achieved. However, achieving and maintaining higher hemoglobin levels carry the risk of hypertension and vascular access thrombosis in dialysis patients and are costly. In addition, a potential for increased risk of death (or no benefit at most) with higher Hb levels has been found in patients with severe cardiac disease in a larger trial. Benefits of and harm from hemoglobin targets should be carefully weighed, and certainly more, proper studies are needed before higher hemoglobin levels (>12 g/dL) are widely adopted in these high-risk patients. PMID- 17136695 TI - Role of prostacyclin (epoprostenol) as anticoagulant in continuous renal replacement therapies: efficacy, security and cost analysis. AB - BACKGROUND: Heparin remains the drug most commonly used for anticoagulation in continuous renal replacement therapies (CRRTs). However, in patients with hypercoagulability, heparin is insufficient or, in cases with an increased risk of bleeding or thrombocytopenia, it may be contraindicated. Epoprostenol, a potent vasodilator, antithrombotic and antiplatelet agent, could be an alternative. PATIENTS AND METHODS: We studied the records of patients treated under continuous venovenous hemodiafiltration in an academic tertiary hospital of 900 beds, between January 2000 and June 2003. Epoprostenol was prescribed to patients with (i) filter hypercoagulability, defined as consumption of 2 or more filters in the last 24 hours; (ii) low platelet count; or (iii) recent severe hemorrhage. RESULTS: Thirty-eight out of 248 (15%) patients who were under CRRT received epoprostenol for more than 72 hours. Epoprostenol was indicated due to filter hypercoagulability in 48%, thrombocytopenia in 68% (7 patients both) and hemorrhage in 3% of cases. The overall time for epoprostenol therapy was 9,749 hours. The mean filter duration previous to epoprostenol was 23 +/- 12 hours and after administering this drug 38.2 +/- 11.9 hours (p = 0.0001). In 6 patients, heparin and epoprostenol were simultaneously administered. The adverse effects were hemorrhage, which presented in 7 patients (18%) and a fall in blood pressure in another 7 (18%), which recovered in the next 24 hour after starting treatment. Cost analysis demonstrates some advantage with epoprostenol in patients with increased tendency to clotting. CONCLUSIONS: Epoprostenol may be safely used to prevent clotting of the extracorporeal circuits, either alone in patients with thrombocytopenia and/or increased risk of bleeding, or in combination with heparin in states of hypercoagulability. PMID- 17136696 TI - A case of essential thrombocytosis developing nephrotic syndrome and severe endothelial damage. AB - We present a 75-year-old Japanese man with essential thrombocytosis presenting nephrotic syndrome. Proteinuria developed soon after the patient was given a diagnosis of essential thrombocytosis and, 4 years later, it increased to a nephrotic range. Renal biopsy revealed one third of the obtained glomeruli totally sclerotic and the other glomeruli showed a marked thickening of the glomerular basement membrane and mild mesangial proliferation. Remarkable widening of the subendothelial space was evident on electron microscopy. Increased expression of platelet derived growth factor receptor beta was detected in the mesangium and interstitium by immunohistochemistry. Abnormal platelet activation in myeloproliferative disease has been shown to contribute in glomerulosclerosis by releasing various growth factors and cytokines including PDGF. Considering his clinical course and the pathological findings, the probable risk factor for developing severe endothelial damage and glomerulosclerosis is due to the persistence of high platelet count and platelet abnormality. PMID- 17136697 TI - Impairment of renal function in Schnitzler's syndrome. AB - The list of multisystem diseases involving both kidney and skin is long and includes immunologic disorders like systemic lupus erythematodes, a broad variety of vasculitides, metabolic disorders like diabetes mellitus, and infectious diseases. The present work describes the first case of renal failure due to Schnitzler's syndrome, a rare entity characterized by the association of generalized chronic urticaria, monoclonal IgM gammopathy, and osteosclerotic lesions. The described patient experiences improvement of renal function after treatment with the chimeric anti-CD20 antibody rituximab, indicating that the impairment of renal function might be mediated by B-lymphocytes. Renal insufficiency is known to be a potential complication of hypocomplementemic urticarial vasculitis. The present case reveals that it can occur in normocomplementemic urticarial vasculitides as well. Schnitzler's syndrome is a rare but probably underdiagnosed syndrome. It should be considered in the differential diagnosis of rash and coincidental renal failure of unknown origin. Immunofixation of immunoglobulins constitutes the crucial diagnostic step, since monoclonal gammopathy is a constant clinical sign in this entity. PMID- 17136698 TI - Nonsurgical treatment of bilateral emphysematous pyelonephritis in a diabetic patient. AB - Bilateral emphysematous pyelonephritis is a rare life-threatening condition affecting almost exclusively patients with diabetes mellitus. Symptoms, which include fever, chills, abdominal and flank pain, nausea, vomiting, dysuria and pyuria, usually mimic those of classic pyelonephritis, and thus clinical suspicion for this urgent condition should be raised in every diabetic patient with similar presentation. Computed tomography (CT) remains the gold standard for the diagnosis demonstrating gas in the renal parenchyma, collecting system or perinephric tissue. Treatment, which should be aggressive, is classically surgical, and early nephrectomy is recommended. Percutaneous drainage associated with medical treatment might be an alternative. Successful exclusively medical treatment has been described but is infrequent and is reserved as an alternative for patients in whom surgical intervention is contraindicated. We report a case of bilateral emphysematous pyelonephritis in an 82-year-old female diabetic patient who presented with symptoms of typical pyelonephritis. Diagnosis was confirmed by CT, and Escherichia coli was identified as the causative factor. The patient was successfully treated medically with intravenous administration of cefepime and amikacin for 14 days and recovered fully. The therapeutical options for this severe but rare condition are discussed. PMID- 17136699 TI - Skin infection due to Alternaria species in kidney allograft recipients: report of a new case and review of the literature. AB - A kidney allograft recipient developed a cutaneous infection 29 months after transplantation, due to the dematiaceous fungus Alternaria infectoria on his right forearm and left leg. Since the lesions were too large to be excised, the patient was treated only with systemic itraconazole and a reduction of the immunosuppressive therapy. After 4 months, the lesions were completely healed, and no relapses were observed at follow-up of 22 months. Twenty-seven other cases of cutaneous alternariosis have been described so far in renal transplant recipients. All types of immunosuppressive treatment can be associated with Alternaria infection, for which predisposing factors are jobs with frequent contact with earth, diabetes mellitus and skin trauma. In 70% of cases the infection occurred within the first year after transplantation. More frequently the lower limbs were involved and the lesions were multiple. Alternaria alternata was the commonest causative agent, followed by Alternaria tenuissima,Alternaria infectoria and Alternaria chartarum. The treatment is far from being standardized, but the best results are obtained with the surgical excision of the lesion(s) associated with systemic antifungal therapy. Since relapses are possible, strict control of the patients over time is essential. PMID- 17136700 TI - Severe cutaneous hypersensitivity to icodextrin in a continuous ambulatory peritoneal dialysis patient. AB - Icodextrin, a glucose polymer, is widely used as an alternative to glucose as the osmotic agent in peritoneal dialysis (PD). We describe a case of a continuous ambulatory peritoneal dialysis patient who developed severe cutaneous hypersensitivity after initiation of icodextrin PD solution. Erythematous skin lesions gradually disappeared after discontinuation of icodextrin PD solution. Although the safety and efficacy of icodextrin PD solution is well documented, clinicians should be mindful of the possibility of severe adverse cutaneous reactions to icodextrin PD solution. PMID- 17136701 TI - Successful pregnancies in dialysis patients including those suffering from cystinosis and familial Mediterranean fever. AB - For women on maintenance dialysis, pregnancy is still uncommon. The outcome of such pregnancies has improved in recent case series. Here, we report in detail the treatment of five successful pregnancies in dialysis patients from our centre. The present case series also includes the first successful pregnancy of a dialysis patient with underlying familial Mediterranean fever, and of a dialysis patient with cystinosis. We treated all patients with an intensified hemodiafiltration protocol, increased erythropoietin dosages, a generous application of water-soluble vitamins and trace elements in addition to a multidisciplinary clinical management approach with a very low threshold for hospital admission. Specifically, we report treatment of arterial hypertension with respect to changes in dry weight and pharmacological therapy. Mean gestational age at delivery was 32.8+/-3.3 weeks and mean birth weight was 1,765+/-554 g. All mothers and newborns were discharged healthy and in good condition. These modified management guidelines have led to a favourable outcome in all our patients including two patients with familial Mediterranean fever and with cystinosis, and may help to guide therapy in other pregnant dialysis patients. PMID- 17136702 TI - Acute deterioration of renal function induced by star fruit ingestion in a patient with chronic kidney disease. AB - Star fruit (carambola) is a popular tropical fruit, usually consumed as fresh fruit or as fruit juice. In patients on dialysis, consumption of star fruits can lead to alterations of consciousness. In this case report, we describe a patient with underlying chronic kidney disease, who developed a rapid increase in serum creatinine and oxalate nephropathy after chronic ingestion of star fruit juice without overt neurotoxicity. The decline in renal function was not fully reversible after stoppage. This case demonstrates that star fruit consumption should be considered as a cause of rapid deterioration in renal function in patients with underlying chronic kidney disease that may result in permanent renal injury. PMID- 17136703 TI - Henoch-Schonlein purpura after postoperative Staphylococcus aureus infection with hepatic IgA nephropathy. AB - A 66-year-old man with a two-year history of hepatitis C viral liver cirrhosis, was diagnosed as having ascending colon cancer. Right hemicolectomy was performed, and a drain was fed down to the anastomosis. On post-operative day (POD) 9, and methicillin-sensitive Staphylococcus aureus (MSSA) was isolated from both drains. After POD 12, relapsing persistent diarrhea with some blood occurred. On POD 20, the temperature increased to 39 degrees C, with symmetrical purpura and swelling in the femurs, and knee arthralgia developed. HSP was suspected. Clinical follow-up showed slight spontaneous reduction of diarrhea and purpura on POD 26. However, despite the negative drain culture, the high fever was maintained on POD 27. Therefore, intravenous steroid pulse therapy was performed. The purpura subsequently disappeared, except for a slight pigmentation and the temperature returned to normal. A renal biopsy was performed 26 days after the appearance of purpura. Pathological views demonstrated acute focal segmental glomerulonephritis-like nephropathy in addition to cirrhotic nephropathy with a membranoproliferative glomerulonephritis (MPGN)-like pattern and the mesangial proliferative glomerulonephritis type. We describe a case of Henoch-Schonlein purpura (HSP) after postoperative Staphylococcus aureus infection of the intra-abdominal drain with IgA nephropathy associated with hepatitis C virus liver cirrhosis. PMID- 17136704 TI - Health utilities in mild cognitive impairment and dementia: a population study in Sweden. AB - OBJECTIVE: To collect new primary data on community-based health utilities (time trade-off values) in different stages of mild cognitive impairment and dementia from a general population sample. METHODS: A cross-sectional study including 1,800 randomly selected members of the Swedish public aged 45-84 was performed through a postal survey; 42% response rate. The Clinical Dementia Rating scale was used for describing progressive stages of dementia in vignettes that were used in combination with time trade-off questions for valuing the perceived quality of life in these stages. RESULTS: The time-trade off values varied considerably across the progressive disease stages. The mean score was 0.82 for mild cognitive impairment, 0.62 for mild dementia, 0.40 for moderate dementia, and 0.25 for severe dementia. In multiple regression analyses, the scores were relatively insensitive to demographic factors like age, gender and self-assessed health. CONCLUSIONS: The results showed that the average time trade-off values declined sharply from mild cognitive impairment to progressing stages of dementia. Since there are many methodological challenges involved in measuring health state utilities in mild cognitive impairment and dementia, more research that evaluates different methods would be welcome. PMID- 17136705 TI - The rate of conversion of mild cognitive impairment to dementia: predictive role of depression. AB - BACKGROUND: Mild cognitive impairment (MCI) is a condition referring to the persons with cognitive deficits measurable in some form or another, but not meeting criteria for dementia, and who have an increased risk of becoming demented. OBJECTIVE: To establish the rate of progression to dementia in MCI, to investigate the risk of conversion for amnestic vs multiple-domains subtypes, and to identify the predictors of progression. METHODS: MCI (n = 105) individuals enrolled in a longitudinal study received annual clinical and psychometric examinations for up to a mean of 3 years. The diagnosis of MCI according to Mayo Clinic Petersen's Criteria was conducted by a panel of specialists. RESULTS: After 3 years of follow-up, 23 of 105 subjects with MCI were diagnosed with dementia. 40 showed cognitive decline not dementia, 34 were stable and showed no cognitive decline or improvement, while eight showed cognitive improvement. CONCLUSIONS: We conclude that conversion rate from MCI to DSM-IIIR dementia was 21.9% over a period of 3 years. The occurrence of depressive symptoms may constitute a predictor for those who are more likely to progress to dementia. The risk of conversion to dementia was higher among the subjects with an evidence of impairment extending beyond memory than with those who suffered only from memory deficits, and the subjects who converted to dementia in this subtype had significantly higher baseline plasma total homocysteine levels than non converters. PMID- 17136706 TI - Clinically significant non-major depression in a community-dwelling elderly population: epidemiological findings. AB - OBJECTIVE: The aim of this study was to investigate the 1-month prevalence, symptom profiles and demographic correlates in late-life clinically significant non-major depression (CSNMD) among a community-dwelling elderly population. METHODS: One thousand five hundred subjects aged 65 years and older, who were randomly selected from three communities in Taiwan, received comprehensive psychiatric assessment by trained psychiatrists. Two categorical diagnoses of depressive disorder, including major depression and CSNMD, were made. The 1-month prevalence was calculated. Frequencies of depressive symptoms across CSNMD and major depression were compared. The risks of CSNMD based on demographic characteristics were estimated using multinominal logistic regression. RESULTS: The 1-month prevalence of CSNMD among the community-dwelling elderly was 8.8%. Symptoms of diminished interest, appetite changes, sleep disturbance, worthlessness or inappropriate guilt, trouble in concentrating or indecisiveness, and suicidal thoughts or acts were less frequent in CSNMD than in major depression, but symptoms of depressed mood, psychomotor changes, and fatigue or loss of energy were as frequent in both categories of illness. CSNMD shares similar demographic risks, such as living in an urban area, female gender and low educational status, with major depression. CONCLUSIONS: CSNMD is common among community-dwelling elders in Taiwan, and with its identical demographic characteristics, but qualitatively different presentation, we suggest CSNMD may be considered part of a spectrum of severe late life depressions with a distinct manifestation. Major depression and CSNMD may share common demographic characteristics with different manifestation. We conclude that late-life depression is a dimensional disease. PMID- 17136707 TI - Subtle imitation behaviour in convenience samples of normal, demented, and currently depressed elderly subjects. AB - BACKGROUND: The clinical significance of imitation behaviour (IB) is unclear. The aim of this study was to investigate the prevalence of subtle naive and obstinate IB in convenience samples of normal elderly, demented, and depressed subjects. METHOD: Subtle IB was assessed using a protocol constructed ad hoc in 146 patients, consecutively referred to a memory clinic having received an ICD-10 diagnosis of either dementia or depression, and in 241 healthy subjects. The prevalence of IB in the three groups was determined and the association with possible demographic, cognitive, and non-cognitive variables analysed. RESULTS: Subtle naive IB was frequent in the elderly with dementia, intermediate in the depressed, and rare in the normal elderly except that the latter frequently stretched out their arms. Obstinate IB never occurred in the normal elderly. IB was predicted by none of the variables used. LIMITATIONS: The groups included were convenience samples with the depressed being a small group precluding further distinction of depressive subtypes. CONCLUSION: Although naive IB is a frequent clinical feature in the demented, it also accompanies depressive disorders in the elderly. It can be observed as context-specific IB in the normal elderly. Obstinate IB does not occur in the normal elderly. PMID- 17136708 TI - The productivity of health care. PMID- 17136710 TI - Reaching the population with dementia drugs: what are the challenges? AB - BACKGROUND: Systematic evidence became available in the late 1990s on efficacy of cholinesterase inhibitors (CHEIs) for patients with mild to moderate Alzheimer's disease (AD) and they began to be used sporadically. Since January 2001 UK based guidelines indicated that one of three cholinesterase inhibitors (CHEIs) could be prescribed for these patients. Since then the cost of prescription in England and Wales has risen. There has been little investigation of uptake at the population level. OBJECTIVE: To estimate the population uptake of CHEIs in a population based study of dementia spanning this period. DESIGN: Using data from a 10-year follow up and a later 12 year interview of the Medical Research Council Cognitive Function and Ageing Study (MRC CFAS), a UK population based longitudinal cohort study of people originally aged 65 years and above, we investigated who was taking CHEIs during the period 2001-2004. We sought information from respondents taking part in the study what medication they were taking on a regular basis. RESULTS: Only 12, of the 219 individuals who received a study diagnosis of dementia were prescribed CHEIs [5%, 95% Confidence Intervals (CI) 3%-9%]) in 2001/2003 and none of the 28 individuals with a study diagnosis of dementia (0%, 95% CI 0-18%) in 2004 were prescribed CHEIs. Uptake was biased towards individuals with more education and higher social class. CONCLUSIONS: These data suggest that any impact on AD progression at the population level will be negligible as prescription of CHEIs and uptake in the age group at highest risk is so limited. There is little evidence that this has changed over time. PMID- 17136711 TI - Effect of indole-3-acetic acid administration by gavage and by subcutaneous injection on rat leukocytes. AB - This study was done to investigate the effect of indole-3-acetic acid (IAA) administered subcutaneously and by gavage on neutrophil function and cytotoxicity in neutrophils and lymphocytes. A gavage administration resulted in an increase in phagocytic capacity in neutrophils in a dose depended manner for 1 mg, 2 mg, 18 mg, and 40 mg of IAA per kg of body mass, respectively, compared with the control. Similarly, subcutaneous administration of IAA at 2, 18, and 40 mg per kg of body mass promoted a significant rise in phagocytosis by neutrophils. H2O2 production in neutrophils from treated rats by gavage was similar to those receiving subcutaneous IAA treatment, and did not show a significant difference between treatments and control. IAA treatment, whether by gavage or subcutaneous, did not produce an alteration in antioxidant enzyme activities or in glucose-6 phosphate dehydrogenase activity of either neutrophils or lymph nodes. Subcutaneous IAA administration did not alter the neutrophil and lymphocyte death as deduced by unaltered membrane integrity, DNA fragmentation and mitochondrial transmembrane potential, compared with controls. In conclusion, IAA administration either subcutaneously or by gavage could increase the phagocytic capacity by neutrophils and this acid administration did not have prooxidant effects or cytotoxic effects on neutrophils and lymphocytes. PMID- 17136712 TI - Behavioural disturbance triggers recognition of dementia by family informants. AB - OBJECTIVE: To determine the frequency of unrecognised dementia in a group of community dwelling elderly, and to identify factors associated with dementia recognition by informants. SAMPLE SELECTION: People over 65 years with an AGECAT case or subcase organic diagnosis or an MMSE < or = 23 were identified from a database of community dwelling elderly. A psychiatrist to confirm the diagnosis of dementia according to ICD-10 criteria interviewed these individuals. STUDY PARTICIPANTS: Sixty-two community dwelling elderly meeting ICD-10 criteria for dementia whom had reliable informants. METHODS: Prior to the start of the interview the informant was questioned about whether they felt the patient had memory difficulties and if so whether they had a medical evaluation for their memory problems. A psychiatrist then interviewed the patient and informant to establish whether that patient met ICD-10 criteria for dementia. Basic sociodemographic details were collected and the following assessments were carried out: the Blessed Dementia Rating Scale, the Clinical Dementia Rating Scale, the Behave-AD and the Baumgarten Behavioural Disturbance Scale. ANALYSIS: Univariate and step-wise forward logistic regression analysis were used to examine the factors associated with recognition of memory difficulties. RESULTS: Twenty-nine percent of family informants of people with dementia failed to recognise a problem with their relatives'memory. Where memory difficulties were recognised only 39% of this group received a medical evaluation. Using univariate analysis recognition of memory difficulties by family informants was associated higher levels of behaviour disturbance ( p = or < 0.0011), greater functional impairment ( p = 0.0039), with increasing cognitive impairment ( p = 0.013). Using a logistic regression model, to test the independence of these variables, increasing behavioural disturbance (p = 0.0001) was associated with recognition of dementia by family informants. CONCLUSIONS: Recognition of memory problems by family members is associated with increasing behavioural disturbance. Even with recognition of dementia, families often fail to seek medical attention. Education of the lay public on the early signs and symptoms of dementia must be a key first step in improving recognition of dementia in the community dwelling elderly. PMID- 17136713 TI - Prevalence of neuropsychiatric symptoms in a large sample of Dutch nursing home patients with dementia. AB - OBJECTIVE: To estimate the prevalence of neuropsychiatric symptoms of dementia patients in Dutch nursing homes. METHODS: Cross-sectional study in a large sample of 1322 demented patients living in 59 dementia special care units (SCUs) in The Netherlands. Symptoms were observed by licensed vocational nurses during regular care-giving in a 2-week observational period prior to assessment. Neuropsychiatric symptoms were assessed using the Neuropsychiatric Inventory- Nursing home version (NPI-NH; frequency X severity score >/= 4) and the Cohen Mansfield Agitation Inventory (CMAI; symptoms occurring at least once a week). RESULTS: More than 80% of these patients suffered from at least one clinically significant symptom, as defined with the NPI-NH frequency X severity score >/= 4. Measured with the NPH-NH agitation/aggression, apathy and irritability were the most frequently observed behaviors, with prevalences of 30-35%. Using the CMAI, 85% of the patients showed at least one symptom of agitation, of which general restlessness was observed most frequently (44%). Other frequently observed symptoms with prevalence rates of 30% were cursing or verbal aggression, constant request for attention, negativism, repetitious sentences, mannerisms, pacing, and complaining. Physically aggressive symptoms such as hitting, kicking, biting occurred less often (less than 13%). CONCLUSIONS: Prevalence rates of neuropsychiatric symptoms in Dutch nursing home patients with dementia residing in SCUs are high, especially agitation and apathy. Insight into the prevalence rates of individual symptoms in patients with dementia has important practical consequences for the accurate planning of staff allotment and stresses the need for patient oriented care. PMID- 17136714 TI - ITP in dynamically double-coated fused-silica capillaries. AB - Bidirectional ITP in fused-silica capillaries double-coated with Polybrene and poly-(vinylsulfonate) is a robust approach for analysis of low-molecular-mass compounds. EOF towards the cathode is strong (mobility >4.0 x 10(-8) m(2)/Vs) within the entire pH range investigated (2.40-8.08), dependent on ionic strength and buffer used and, at constant ionic strength, higher at alkaline pH. Electrokinetic separations and transport in such coated capillaries can be described with a dynamic computer model which permits the combined simulation of electrophoresis and electroosmosis in which the EOF is predicted either with a constant (i.e. pH- and ionic strength-independent) or a pH- and ionic strength dependent electroosmotic mobility. Detector profiles predicted by computer simulation agree qualitatively well with bidirectional isotachopherograms that are monitored with a setup comprising two axial contactless conductivity detectors and a UV absorbance detector. The varying EOF predicted with a pH- and ionic strength-dependent electroosmotic mobility can be regarded as being realistic. PMID- 17136715 TI - Exploring the evaluation of net charge, hydrodynamic size and shape of peptides through experimental electrophoretic mobilities obtained from CZE. AB - This work explores the validity of simple CZE models to analyze the electrophoretic mobilities of 102 peptides reported in literature. These models are based mainly on fundamental physicochemical theories providing analytical expressions amenable to relatively simple numerical analysis. Thus, the Linderstrom-Lang capillary electrophoresis model (LLCEM) and its perturbed version (PLLCEM), proposed and applied previously to the CZE of globular proteins, are adapted and used here for peptides. Also the effects of pK-shifts on net charge, hydration and hydrodynamic size and shape of peptides are analyzed and discussed. Emphasis is placed on the fact that these parameters are physically coupled, and thus a variation in the net charge may produce an appreciable change in the hydrodynamic size of peptides. Within the framework of CZE, peptides may be assumed as having a hydrodynamic volume associated with either spherical or spheroidal particles. The effects on peptide net charge and hydrodynamic size, of electrostatic interaction between a pair of charged groups in the chain and electrical permitivitty around the peptide domain are studied. The predictions of the PLLCEM and LLCEM are in good agreement with results reported previously in the literature. Several limitations concerning these models and some needs for further research are also described. PMID- 17136716 TI - Universal method for synthesis of artificial gel antibodies by the imprinting approach combined with a unique electrophoresis technique for detection of minute structural differences of proteins, viruses, and cells (bacteria). III: gel antibodies against cells (bacteria). AB - Artificial antibodies in the form of gel granules were synthesized from the monomers acrylamide and N,N'-methylenebisacrylamide by the imprinting method in the presence of Echerichia coli bacteria as template. The electrophoretic migration velocities of the gel antibodies (i) saturated with the antigen (Escherichia coli MRE-600), (ii) freed of the antigen, and (iii) resaturated with bacteria, were determinated by electrophoresis in a rotating narrow-bore tube of 245 mm length and the 2.5 and 9.6 mm inner and outer diameters, respectively. Removal of bacteria from the gel antibodies was made by treatment with enzymes, followed by washing with SDS and buffer. Gel granules becoming charged by adsorption of bacteria move in an electrical field. We obtained a significant selectivity of gel antibodies for E. coli MRE-600, since the granules did not interact with Lactococcus lactis; and when E. coli BL21 bacteria were added to the gels selective for E. coli MRE-600, a significant difference in the migration rate of the complexes formed with the two strains was observed indicating the ability of differentiation between the two strains. The gel antibodies can be used repeatedly. The new imprinting method for the synthesis of artificial gel antibodies against bioparticles described herein, and the classical electrophoretic analysis technique employed, thus represent - when combined - a new approach to distinguish between different types and strains of bacteria. The application area can certainly be extended to cover other classes of cells. PMID- 17136717 TI - Combining stir bar sorptive extraction and MEKC for the determination of polynuclear aromatic hydrocarbons in environmental and biological matrices. AB - In this work, stir bar sorptive extraction and liquid desorption was combined with MEKC and diode-array detection (SBSE-LD-MEKC-DAD) for the determination of polynuclear aromatic hydrocarbons (PAHs) in aqueous medium, using biphenyl, fluorene, anthracene, phenanthrene, fluoranthene and pyrene as model compounds. MEKC-DAD conditions and parameters affecting SBSE-LD efficiency are fully discussed. Assays performed on aqueous samples spiked at trace levels, yielded recoveries ranging from 55.5 +/- 6.1% (pyrene) to 70.7 +/- 7.1% (anthracene), under optimized experimental conditions. The methodology proved to be nearly described by the octanol-water partition coefficients (K(PDMS/W) approximately K(O/W)). The analytical performance showed good precision (<12.0%), suitable detection limits (2-11 microg/L) and convenient linear dynamic ranges (r(2)>0.99) from 5 to 25 microg/L for anthracene and 25 to 125 microg/L for the remaining compounds. The application of the proposed methodology to environmental water, sediments and fish bile matrices demonstrated good selectivity and accuracy. SBSE LD combined with MEKC-DAD was shown to be an easy, reliable and robustness methodology, as well as a good analytical alternative to monitor environmental priority pollutants. PMID- 17136718 TI - Chiral resolution of tryptophan derivatives by CE using canine serum albumin and bovine serum albumin as chiral selectors. AB - This work deals with the application of BSA and canine serum albumin (CSA) for enantioseparation of tryptophan derivatives with CE. The aim of this work was the investigation of the influence of different functional groups of tryptophan derivatives on enantioseparation. CSA as a chiral selector was tested to compare its selector properties with those of BSA. The enantiomers of the tryptophan derivatives were separated by adding BSA or CSA to the BGE. The influence of pH, temperature, BSA and CSA concentration and organic modifiers was investigated. It was found that the stereoselectivity for the different tryptophan derivatives is dependent on the albumin species. It turned out that the different functional groups of the derivatives showed a significant influence on stereoselectivity. PMID- 17136719 TI - CE for cytokinin analyses: a review. AB - Analyses of cytokinins are very important in both plant physiological and biomedical research as they are implicated in many biological processes. Reliable, sensitive, selective and inexpensive methods that are flexible and designed for automation are required for these analyses. This review addresses the advances made in the separation and determination of cytokinins by CE as well as the other applications of CE (i.e., determination of dissociation constants and complexation constants of cytokinins). The various CE modes used to separate the compounds and the quantification strategies are examined. Special attention is also focused on those aspects that improve on the sensitivity and/or selectivity, such as sample extraction and preconcentration, on-line preconcentration techniques (stacking), and/or specific detectors (e.g., MS). With the coupling to the preconcentration techniques and certain detection systems, numerous CE methods can potentially be adapted for the analysis of cytokinins in complex biological samples. Therefore, we would anticipate wider applications of CE methods in the near future for cytokinin analyses, which should facilitate a decrease in analysis cost and should help to improve analysis efficiency. PMID- 17136720 TI - Optimization of CZE for analysis of phytochemical bioactive compounds. AB - Advantages of CZE such as high efficiency, low cost, short analysis time, and easy implementation result in its wide applications for analysis of phytochemical bioactive compounds (e.g. flavonoids, alkaloids, terpenoids, phenolic acid, saponins, anthraquinones and coumarins). However, several aspects, including sample preparation, separation, and detection have significant effects on CZE analysis. Therefore, optimization of these procedures is necessary for development of the method. In this review, sample preparation such as extraction method and preconcentration, separation factors including buffer type, concentration and pH, additives, voltage and temperature, as well as detection, e.g. direct and indirect UV detection, LIF and MS were discussed for optimization of CZE analysis on phytochemical bioactive compounds. The optimized strategies were also reviewed. PMID- 17136721 TI - Analysis of selected constituents in methanolic extracts of Hypericum perforatum collected in different localities by capillary ITP-CZE. AB - The on-line combination of CZE with capillary ITP (ITP-CZE) was used for the separation and quantification of selected flavonoids and phenolic acids in Hypericum perforatum leaves and flowers collected in six different localities in Slovakia. The leading electrolyte in the ITP preseparation step was 10 mM HCl with Tris as counterion (pH* 7.2). The terminating electrolyte was 50 mM boric acid of pH* 8.2 (adjusted with barium hydroxide). The BGE in the electrophoretic step contained 25 mM beta-hydroxy-4-morpholinopropanesulfonic acid (MOPSO), 50 mM Tris, 65 mM boric acid, pH* 8.3. The content of methanol in all electrolytes was 20% v/v. The total time of the analysis (including the preseparation step) was approximately 35 min. The rectilinear calibration ranges were between 0.125 and 5.0 microg/mL with kaempferol as internal standard. The correlation coefficients ranged between 0.9912 (for quercitrin and chlorogenic acid) and 0.9988 (for isoquercitrin). The RSD values are between 0.86 and 7.78% (n = 6) when determining rutin and quercetin (4 microg/mL). The optimized method was employed for the assay of flavonoids in medicinal plant extract of different collections of Hypericum perforatum haulm. The variability of the content of the active components depending on the place of collection was confirmed. PMID- 17136722 TI - Facile separation and determination of Aconitine alkaloids in traditional Chinese medicines by CE with tris(2,2'-bipyridyl) ruthenium(II)-based electrochemiluminescence detection. AB - A facile CE method coupled with tris(2,2'-bipyridyl) ruthenium(II)-based electrochemiluminescence [Ru(bpy)(3) (2+)] detection was developed for simultaneous determination of Aconitum alkaloids, i.e., hypaconitine (HA), aconitine (AC), and mesaconitine (MA) in baseline separation. The optimal separation of these Aconitum alkaloids was achieved in a fused-silica capillary column (50 cm x 25 microm id) with 30 mM phosphate solution (pH 8.40) as running buffer at 12 kV applied voltage. The three alkaloids can be determined within 10 min by a single run. The calibration curves showed a linear range from 2.0 x 10( 7) to 2.0 x 10(-5) M for HA, 3.4 x 10(-7) to 1.7 x 10(-5) M for AC, and 3.8 x 10( 7) to 1.9 x 10(-5) M for MA. The RSDs for all analytes were below 3.01%. Good linear relationships were found with correlation coefficients for all analytes exceeding 0.993. The detection limits were 2.0 x 10(-8) M for HA, 1.7 x 10(-7) M for AC, and 1.9 x 10(-7) M for MA under optimal conditions. This method was successfully applied to determine the three alkaloids in Aconitum plants. PMID- 17136724 TI - Determination of trace cationic impurities in butylmethylimidazolium-based ionic liquids: from transient to comprehensive single-capillary counterflow isotachophoresis-zone electrophoresis. AB - Determination of impurities in ionic liquids (ILs) remains a difficult task. In this work, the hyphenation of isotachophoretic (ITP) preconcentration to zone electrophoresis (ZE) has been explored for the trace analysis of the cationic impurities Na(+), Li(+), and methylimidazolium (MI(+)) in butylmethylimidazolium (BMI(+))-based ILs. Simultaneous detection of UV-transparent and UV-absorbing impurities was ensured by a BGE composed of creatinine-acetate buffer. To induce ITP, three different strategies were evaluated: (i) Sample self-stacking ensured by the addition of ammonium acetate (NH(4)Ac) to 25-50-fold diluted IL solution (transient ITP). (ii) Complete ITP-ZE separation performed in a single capillary: ITP was realized in discontinuous electrolytes comprising an 80 mM NH(4)Ac, 40 mM acetic acid, 30 mM alpha-CD, pH 5.05, leading electrolyte (LE) and a 10 mM creatinine, 10 mM acetic acid, pH 4.9, terminating electrolyte (TE). To create the ZE stage, the ITP stack of analytes was moved back toward the capillary inlet by pressure and simultaneously the capillary was filled with the BGE. This protocol made it possible to accommodate a 2.5-times diluted IL sample. (iii) Complete counterflow ITP-ZE with continuous electrokinetic sample supply: the ITP stage was performed in a capillary filled with a 150 mM NH(4)Ac, 75 mM acetic acid, 30 mM alpha-CD, pH 5.0 LE, with 40-times diluted IL at the capillary inlet. BMI(+) from IL acts as the terminating ion. The LODs reached in this latter case were at the 10 and 1 ppb levels for MI(+) and Li(+) in diluted IL matrix, respectively. PMID- 17136727 TI - The use of CE-electrochemiluminescence with ionic liquid for the determination of bioactive constituents in Chinese traditional medicine. AB - CE / tris(2,2-bipyridyl) ruthenium(II) (Ru(bpy)(3) (2+)) electrochemiluminescence (ECL), CE-ECL, with an ionic liquid (IL) detection system was established for the determination of bioactive constituents in Chinese traditional medicine opium poppy which contain large amounts of coexistent substances. A minimal sample pretreatment which involves a one-step extraction approach avoids both sample loss and environmental pollution. As the nearby hydroxyl groups in some alkaloid such as morphine may react with borate to form complexes and IL, as a high conductivity additive in running buffer, could cause an enhanced field-amplified effect of electrokinetic injection. Running buffer containing 25 mM borax-8 mM 1 ethyl-3-methylimidazolium tetrafluoroborate (EMImBF(4))IL (pH 9.18) was used which resulted in significant changes in separation selectivity and obvious enhancement in ECL intensities for those alkaloids with similar structures. Sensitive detection could be achieved when the distance between the Pt working electrode and the outlet of separation capillary was set at 150 microm and the stainless steel cannula was fixed approximately 1 cm away from the outlet of the capillary. Quantitative analysis of four alkaloids was achieved at a detection voltage of 1.2 V and a separation voltage of 15 kV in less than 7 min. Detection limits of thebaine, codeine, morphine, and narcotine were 2.5 x 10(-7), 2.5 x 10( 7), 1 x 10(-9) and 1 x 10(-6) M(S/N = 3), respectively. The method was successfully applied to determine the amounts of opium alkaloids in real poppy samples. PMID- 17136728 TI - Mass distribution, polydispersity and focusing properties of carrier ampholytes for IEF II: pH 4-6 intervals. AB - For studying the M(r) distribution and number of species in narrow-range (2 pH unit wide, in the nominal pI 4-6 interval) carrier ampholytes from four commercial sources (Bio-Lyte, Servalyt, Ampholine and Pharmalyte), a 2-D technique was adopted consisting of a focusing step in a liquid phase (Rotofor, yielding 20 fractions) followed by orthogonal CE in both, acidic and basic buffers. As a final step, every other fraction was analyzed by CE-MS. The findings: Ampholine contains 80 different M(r) compounds, in the M(r) interval 203 to 893 Da, for a total of 325 isoforms. Bio-Lyte consists of 66 different M(r) species, in the M(r) range 388 to 835 Da, for a total of 436 isoforms. Servalyt is made of 199 different M(r) compounds, in the M(r) interval 204 to 907 Da, for a total of 1302 isoforms. Pharmalyte pH 4-6.5, comprises 217 amphoteres, in the M(r) range 150 to 1179 Da, for a total of 812 isoforms. Pharmalyte appears to be the best brand, with the vast majority of species focusing sharply at their pI position and <5% "poor" species, distributed along the entire pH gradient, denoting an extremely shallow pH/mobility curve across the pI value. PMID- 17136729 TI - Comparison of the hydrolysis of polyethylene terephthalate fibers by a hydrolase from Fusarium oxysporum LCH I and Fusarium solani f. sp. pisi. AB - The hydrolysis of polyethylene terephthalate (PET) fibers by two fungal hydrolases was investigated. The hydrolase from a newly isolated Fusarium oxysporum strain (LCH 1) was more efficient in releasing terephthalic acid from PET fibers compared to the enzyme from F. solani f. sp. pisi DSM 62420 when equal amounts of p-nitrophenyl butyrate-hydrolyzing activity were employed. PET fabrics treated under the same conditions with the enzyme from F. oxysporum LCH 1 also showed a considerably higher increase in hydrophilicity compared to fabrics treated with the enzyme from F. solani f. sp. pisi DSM 62420. PMID- 17136730 TI - Bioreactor technology: a novel industrial tool for high-tech production of bioactive molecules and biopharmaceuticals from plant roots. AB - Plants are the richest source for different bioactive molecules. Because of the vast number of side effects associated with synthetic pharmaceuticals, medical biotechnologists turned to nature to provide new promising therapeutic molecules from plant biofactories. The large-scale availability of the disease- and pesticide-free raw material is, however, restricted in vivo. Many bioactive plant secondary metabolites are accumulated in roots. Engineered plants can also produce human therapeutic proteins. Vaccines and diagnostic monoclonal antibodies can be won from their roots, so that engineered plants hold immense potential for the biopharmaceutical industry. To obtain sufficient amounts of the plant bioactive molecules for application in human therapy, adventitious and hairy roots have to be cultured in in vitro systems. High-tech pilot-scale bioreactor technology for the establishment of a long-term adventitious root culture from biopharmaceutical plants has recently been established. In this review, I briefly discuss a technology for cultivating bioactive molecule-rich adventitious and hairy roots from plants using a high-tech bioreactor system, as well as the principles and application of genome-restructuring mechanisms for plant-based biopharmaceutical production from roots. High-tech bioreactor-derived bioactive phytomolecules and biopharmaceuticals hold the prospect of providing permanent remedies for improving human well-being. PMID- 17136731 TI - Impact of denaturation with urea on recombinant apolipoprotein A-IMilano ion exchange adsorption: equilibrium uptake behavior and protein mass transfer kinetics. AB - We have studied the equilibrium uptake behavior and mass transfer rate of recombinant apolipoprotein A-I(Milano) (apo A-I(M)) on Q Sepharose HP under non denaturing, partially denaturing, and fully denaturing conditions. The protein of interest in this study is composed of amphipathic alpha helices that serve to solubilize and transport lipids. The dual nature of this molecule leads to the formation of micellar-like structures and self association in solution. Under non denaturing conditions equilibrium uptake is 134 mg/mL media and the isotherm is essentially rectangular. When fully denatured with 6 M urea, the equilibrium binding capacity decreases to 25 mg/mL media and the isotherm becomes less favorable. The decrease in both binding affinity and media capacity when the protein is completely denatured with 6 M urea can be explained by the loss of all alpha helical structure. The rate of apo A-I(M) mass transfer on Q Sepharose HP was characterized using a macropore diffusion model. Results of modeling studies indicate that effective pore diffusivity increases from 4.5 x 10(-9) cm2/s in the absence of urea to 6.0 x 10(-8) cm2/s when apo A-I(M) is fully denatured with 6 M urea. Based on light-scattering data reported for apo A-I, protein self association appears to be the dominant cause of slow protein mass transfer observed under non-denaturing conditions. PMID- 17136732 TI - Enzymes fight chemical weapons. PMID- 17136733 TI - Utilization of renewables for lactic acid fermentation. AB - Originally, lactic acid was produced from pure substrates like glucose. Increasingly, however, agricultural feedstocks such as grains and green biomass are also being used as raw materials for the biotechnological production of lactic acid. A high-productivity lactic acid bacterium strain was selected, process parameters were optimized for the batch fermentation on a laboratory scale, and its performance at cultivation on a barley hydrolysate medium together with different supplements was examined. The present results for the cultivation of the Lactobacillus paracasei on complex nutrient broth are in the same range as those for another strain of the same species with pure glucose, de Man, Rogosa and Sharpe medium (MRS) minerals, peptone and yeast extract. Under these conditions, this strain was able to accumulate more than 100 g lactate/L in the MRS medium. Medium optimization experiments showed that the main part of the nitrogen-containing nutrients in the medium (peptone, yeast extract) can be replaced by protein extracts from green biomass (lucerne green juice). The green juice after pressing fresh biomass contains a series of nitrogen-containing compounds and inorganic salts, which are essential for cell growth. Thus, on laboratory scale, we have demonstrated that it is possible to substitute synthetic nutrients by renewable resources like cereals and green biomass without any loss of productivity. This high biomass concentration together with the number of living cells could increase the productivity to higher levels compared to the well-adapted synthetic nutrients of MRS. PMID- 17136735 TI - LIF detection of peptides and proteins in CE. AB - CE- and microchip-based separations coupled with LIF are powerful tools for the separation, detection and determination of biomolecules. CE with certain configurations has the potential to detect a small number of molecules or even a single molecule, thanks to the high spatial coherence of the laser source which permits the excitation of very small sample volumes with high efficiency. This review article discusses the use of LIF detection for the analysis of peptides and proteins in CE. The most common laser sources, basic instrumentation, derivatization modes and set-ups are briefly presented and special attention is paid to the different fluorogenic agents used for pre-, on- and postcapillary derivatization of the functional groups of these compounds. A table summarizing major applications of these derivatization reactions to the analysis of peptides and proteins in CE-LIF and a bibliography with 184 references are provided which covers papers published to the end of 2005. PMID- 17136736 TI - Recent progress in polar stationary phases for CEC. AB - This review summarizes most of the recent developments in the preparation and application of polar stationary phases for CEC covering the literature published since the year 2004. These polar stationary phases have been adopted for separation of analytes by the modes of packing column CEC, open-tubular CEC (o CEC) and monolithic column CEC. Currently, development of o-CEC using biomolecules, such as protein and DNA, as the immobilized ligands is highlighted partly due to the simplicity of preparation. Furthermore, monolithic columns have been extended quickly, particularly inorganic materials-based monoliths, such as silica, zirconia, hafnium, etc., as an alternative to packed columns have been developed quickly. PMID- 17136737 TI - Recent progress in capillary ITP. AB - ITP has been attracting constant attention for many years due to its principal capability to concentrate trace analytes by several orders of magnitude. In the current capillary format, it is able to concentrate trace analytes diluted to several microliters of an original sample into concentrated zones having volumes in the range of picoliters. Due to this reason, ITP holds an important position in many current multistage and multidimensional separation schemes. This article links up previous reviews on the topic and summarizes the progress of analytical capillary ITP since 2002. Almost 100 papers are reviewed that include methodological novelties, instrumental aspects, and analytical applications. Papers using ITP and/or isotachophoretic principles as part of multistage and/or multidimensional separation schemes are also included. PMID- 17136738 TI - Contemporary sample stacking in CE: a sophisticated tool based on simple principles. AB - Sample stacking is a general term for methods in CE which are used for on-line concentration of diluted analytes. During the stacking process, analytes present at low concentrations in a long injected sample zone are concentrated into a short zone (stack). The stacked analytes are then separated and individual zones are detected. Thus stacking provides better separation efficiency and detection sensitivity. Many papers have been published on stacking till now, various procedures have been described, and, many names have been proposed for stacking procedures utilizing the same principles. This contribution brings an easy and unified view on stacking, describes the basic principles utilized, makes a list of recognized operational principles and brings an overview of principal current procedures. Further, it surveys selected recent practical applications ordered according to their operational principles and includes the terms, nicknames, and acronyms used for these actual stacking procedures. This contribution may help both newcomers and experts in the field of CE to orient themselves in the already quite complex topic of sample stacking. PMID- 17136739 TI - Advances in chiral separation using capillary electromigration techniques. AB - This review gives an overview of recent developments in CZE, EKC, and CEC covering the literature since the year 2004. Since there appeared a special issue on applications, this review focuses on the progress in electromigration techniques and new methodological developments. New techniques, new chiral selectors as well as new chiral stationary phases for CEC are discussed. PMID- 17136740 TI - Recent progress in the use of ionic polymers as pseudostationary phases for EKC. AB - This review concerns the introduction, characterization, and application of polymeric pseudostationary phases (PSPs) for EKC since 2004. Achiral and chiral polymers and separations are reviewed, as is the application of polymeric PSPs for the combination of EKC with mass spectrometric detection. PMID- 17136741 TI - Recent advances in the development and application of microemulsion EKC. AB - Microemulsion EKC (MEEKC) is an electrodriven separation technique. Separations are typically achieved using oil-in-water microemulsions, which are composed of nanometre-sized oil droplets suspended in an aqueous buffer. The droplets are stabilised by a surfactant and a cosurfactant. The novel use of water-in-oil microemulsions has also been investigated. This review summarises the advances in the development of MEEKC separations and also the different areas of application including determination of log P values, pharmaceutical applications, chiral analysis, natural products and bioanalytical separations and the use of new methods such as multiplexed MEEKC and high speed MEEKC. Recent applications (2004 2006) are tabulated for each area with microemulsion composition details. PMID- 17136742 TI - MEKC: an update focusing on practical aspects. AB - This paper reviews recent methodological and instrumental advances in MEKC. Improvements in sensitivity arising from the use of on-line sample concentration (sweeping, stacking, and combination of both protocols) and derivatization (in capillary reactions and coupling with flow-injection systems) and improvements in resolution obtained by changing the composition of the BGE (e.g., with organic modifiers, ionic liquids, nonionic and zwitterionic surfactants, mixed micelles, and vesicles) or using coated capillaries are discussed in detail. In addition, MS and LIF spectroscopy are examined in relation to their advantages and restrictions as applied to MEKC analysis. Some thoughts on potential future directions are also expressed. PMID- 17136743 TI - A versatile esterase from Bacillus subtilis: cloning, expression, characterization, and its application in biocatalysis. AB - An esterase from Bacillus subtilis DSM402 (BS2) was cloned and functionally expressed in E. coli. The enzyme is active up to 50 degrees C, and the V(max) (1449 mM/min) and K(M) values (119 mM) were determined using p-nitrophenyl acetate as substrate. BS2 belongs to the few hydrolases that can act on tertiary alcohols and was therefore used to resolve racemic acetates of selected tertiary alcohols, but also to selectively remove the tert-butyl ester protecting group from peptides. In addition, the enzyme shows promiscuous amidase activity. PMID- 17136746 TI - Developments in cluster randomized trials and Statistics in Medicine. AB - The design and analysis of cluster randomized trials has been a recurrent theme in Statistics in Medicine since the early volumes. In celebration of 25 years of Statistics in Medicine, this paper reviews recent developments, particularly those that featured in the journal. Issues in design such as sample size calculations, matched paired designs, cohort versus cross-sectional designs, and practical design problems are covered. Developments in analysis include modification of robust methods to cope with small numbers of clusters, generalized estimation equations, population averaged and cluster specific models. Finally, issues on presenting data, some other clustering issues and the general problem of evaluating complex interventions are briefly mentioned. PMID- 17136749 TI - Molecular simulation of protein aggregation. AB - Computer simulation offers unique possibilities for investigating molecular-level phenomena difficult to probe experimentally. Drawing from a wealth of studies concerning protein folding, computational studies of protein aggregation are emerging. These studies have been successful in capturing aspects of aggregation known from experiment and are being used to refine experimental methods aimed at abating aggregation. Here we review molecular-simulation studies of protein aggregation conducted in our laboratory. Specific attention is devoted to issues with implications for biotechnology. PMID- 17136751 TI - The notochord in the mammalian adult: a paradox. PMID- 17136752 TI - Immunosuppressive medications and hospitalization for cardiovascular events in patients with rheumatoid arthritis. AB - OBJECTIVE: The risk of cardiovascular disease (CVD) is increased in patients with rheumatoid arthritis (RA), most likely because of increased systemic inflammation. Prior research suggests that immunosuppressive medications may reduce the risk of CVD among RA patients. This study was undertaken to investigate the effects of various immunosuppressive medications on the risk of cardiovascular events among a group of older patients with RA. METHODS: In this nested case-control study, the source cohort was derived from Medicare beneficiaries receiving a drug benefit from the state of Pennsylvania. These individuals were required to have been diagnosed as having RA on at least 2 visits and to have filled a prescription for an immunosuppressive agent. Cases were defined as those patients who were hospitalized for a cardiovascular event such as myocardial infarction or stroke, and 10 control subjects were matched to each case by age, sex, and calendar year of the index date (the time of the first cardiovascular event in each case). Current use of an immunosuppressive medication was defined as having filled a prescription for these agents within the 90 days prior to the index date. Multivariate logistic regression models that included important covariates were assessed to determine the risk of cardiovascular events associated with immunosuppressive agents and their combinations. RESULTS: Among the study cohort, we identified 3,501 RA patients who fulfilled our eligibility criteria. During followup of this cohort, 946 patients were hospitalized for a cardiovascular event. Although the 95% confidence intervals (95% CIs) were wide in adjusted risk regression models with methotrexate (MTX) monotherapy as the reference group, biologic immunosuppressive agents showed neither protective nor deleterious effects (with biologics monotherapy, odds ratio [OR] 1.0, 95% CI 0.5-1.9; with biologics plus MTX combination therapy, OR 0.8, 95% CI 0.3-2.0; and with biologics plus other immunosuppressive agents, OR 1.2, 95% CI 0.7-2.2). Monotherapy with oral glucocorticoids was associated with an increased risk of cardiovascular events (OR 1.5, 95% CI 1.1-2.1), and a similar trend in the direction of risk was seen with glucocorticoid combination therapy (OR 1.3, 95% CI 0.8-2.0). Cytotoxic immunosuppressive agents other than MTX (azathioprine, cyclosporine, and leflunomide) were also associated with an increased risk of cardiovascular events (with both monotherapy and combination treatment, OR 1.8, 95% CI 1.1-3.0). CONCLUSION: When compared with RA patients receiving MTX monotherapy, those receiving biologic immunosuppressive agents had neither an increased nor decreased risk of experiencing a cardiovascular event, whereas use of oral glucocorticoids and cytotoxic immunosuppressive agents was associated with significant increases in the risk of cardiovascular events. PMID- 17136753 TI - Nucleus pulposus notochord cells secrete connective tissue growth factor and up regulate proteoglycan expression by intervertebral disc chondrocytes. AB - OBJECTIVE: To identify the components of conditioned medium obtained from intervertebral disc nucleus pulposus-derived canine notochord cells, and to evaluate the capacity of such factors to affect disc-derived chondrocyte gene expression of aggrecan, versican, and hyaluronic acid synthase 2 (HAS-2) as a function of culture conditions. METHODS: Canine notochord cells obtained from nonchondrodystrophic dogs were cultured within alginate beads under conditions of serum deficiency (Dulbecco's modified Eagle's medium [DMEM]) to produce notochord cell-conditioned medium (NCCM). NCCM was evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and liquid chromatography-tandem mass spectroscopy. Bovine disc-derived chondrocytes were cultured with serum-deficient medium (DMEM) and NCCM and assayed for the effect of tissue culture conditions on aggrecan, versican, and HAS-2 gene expression. Next, chondrocyte gene expression for aggrecan was evaluated using DMEM containing recombinant connective tissue growth factor (rCTGF), and the results compared with those obtained using NCCM and DMEM. RESULTS: NCCM contained aggrecan, Cu/Zn superoxide dismutase, fibronectin, and CTGF precursor. Culture with NCCM caused an up-regulation of aggrecan, versican, and HAS-2 gene expression. NCCM induced aggrecan gene expression in chondrocytes at a level similar to that induced by 100-200 ng/ml rCTGF. Nonchondrodystrophic and chondrodystrophic canine notochord cells exhibited similar levels of CTGF gene expression. CONCLUSION: Nucleus pulposus derived notochord cells secrete CTGF (CCN2), a recently discovered multifunctional growth factor. There is no difference between CTGF gene expression in nonchondrodystrophic and chondrodystrophic canine notochord cells, suggesting a possible role of CTGF as an anabolic factor within the disc nucleus that is, to at least some degree, dependent on the population of notochord cells within the disc nucleus. PMID- 17136754 TI - Partial versus full PTSD in the Korean community: prevalence, duration, correlates, comorbidity, and dysfunctions. AB - A few studies have been conducted on the prevalence of partial posttraumatic stress disorder (PTSD), but points of agreement and disagreement between full and partial PTSD have not been fully investigated. We interviewed a representative sample of 6,258 subjects, ages 18-64 years, in household visits using the Korean version of the Composite International Diagnostic Interview (K-CIDI 2.1). "Partial PTSD" was defined as >/=1 symptom in each of the three symptom groups (Criteria B, C, and D) and duration of >/=1 month (Criterion E). Estimated lifetime prevalence of partial PTSD was 2.7%, and that of full PTSD was 1.7%. A "female gender" risk factor was significantly associated with both partial and full PTSD. The mean duration of partial PTSD was 6.5 years, which was not significantly different from the 5.7 year duration of full PTSD. Traumas associated with the development of partial rather than full PTSD were "natural disaster with fire" and "military combat" in men, and "witnessing a traumatic situation" and "learning about traumas to others" in women, whereas "threatened by others" was more associated with the development of full PTSD. The rates of multiple comorbid disorders and of comorbid major depressive disorder and dysfunctions in work during the 1-month period prior to interview did not differ significantly between the partial and full PTSD groups. In conclusion, partial PTSD did not differ significantly from full PTSD in terms of duration, comorbidity, and dysfunction, but they differed markedly in terms of associated trauma types. PMID- 17136755 TI - Anxiety sensitivity and CO2 challenge reactivity as unique and interactive prospective predictors of anxiety pathology. AB - Emerging evidence suggests that anxiety sensitivity (AS) and CO(2)-induced fear reactivity are associated with panic attacks and anxiety disorders. However, evidence regarding the unique and potentially synergistic effects of these variables is currently lacking. Our primary aims in this study were to determine whether AS and CO(2)-induced fear reactivity are unique and potentially interactive vulnerability factors involved in the pathogenesis of panic attacks and anxiety psychopathology. A large nonclinical sample of young adults (N=404) was prospectively followed over approximately 2 years. AS (i.e., 16-item Anxiety Sensitivity Index total scores) and biological challenge reactivity [i.e., fearful responding to pre- and postchallenge changes in subjective units of distress (SUDS) to a 20-s 20% CO(2) challenge] at study entry served as the primary predictor variables. Consistent with expectation, AS and challenge reactivity correlated only moderately with one another. Challenge reactivity was uniquely associated with the development of spontaneous panic attacks, whereas AS was uniquely associated with anxiety disorder diagnoses, including panic disorder. Moreover, the combination of both risk factors predicted spontaneous panic attacks beyond the effects of either risk factor individually. These data provide novel evidence for the unique and combined effects of AS and CO(2) induced fear reactivity as risk factors in the development of anxiety and its disorders. PMID- 17136756 TI - Notice of Retraction: "Comparison of histological grading and large-scale genomic status (DNA ploidy) as prognostic tools in oral dysplasia" (J Pathol 2001;194:303 310). PMID- 17136757 TI - Epithelial-mesenchymal interactions in keloid pathogenesis modulate vascular endothelial growth factor expression and secretion. AB - Vascular endothelial growth factor (VEGF) plays an important role in angiogenesis during the wound healing process. As epithelial-mesenchymal interactions have been shown to regulate a plethora of genes in wound healing, we hypothesized that these interactions might have a role in modulating VEGF expression and angiogenesis. A two chamber co-culture model was used, wherein normal and keloid keratinocytes and fibroblasts were physically separated by membrane inserts while allowing cytokine diffusion. Cell lysates obtained from keratinocytes co-cultured with fibroblasts demonstrated increased expression of VEGF. An enzyme-linked immunosorbent assay (ELISA) showed significant increase in VEGF expression in co culture conditioned media compared with controls. Additionally, the conditioned medium from keloid keratinocyte and fibroblast co-cultures increased proliferation and formation of complex three-dimensional capillary-like structures in human umbilical vein endothelial cells, emphasising the importance of epithelial-mesenchymal interactions in the angiogenic process. Immunostaining of keloid tissue localized VEGF in the basal layer of the epidermis and also demonstrated higher blood vessel density than normal skin. Keloid tissue extract also demonstrated increased expression of VEGF compared with normal skin. It is likely that epidermal VEGF exerts significant paracrine control over the dynamics and expression profile of underlying dermal fibroblasts. Addition of the inhibitors WP631, mitoxantrone, and Rapamycin to keloid keratinocyte and fibroblast co-cultures, downregulated secreted VEGF expression in a dose dependent manner, suggesting therapeutic potential for these compounds in the treatment of keloid scars. PMID- 17136758 TI - The C allele of the GNB3 C825T polymorphism of the G protein beta3-subunit is associated with an increased risk for the development of oncocytic thyroid tumours. AB - Carriers of the C allele of the common C825T polymorphism in the GNB3 gene of the G protein have been associated with the development of follicular thyroid adenomas. Since the C allele of this polymorphism is related to a lower signalling capacity, it was speculated whether the C825T polymorphism may play a particular role in oncocytic thyroid tumours, which are recognized for their reduced ability to synthesize thyroid-specific proteins and hormones, although they possess an intact thyroid-stimulating hormone receptor-adenylyl cyclase system. Using pyrosequencing, both the genotype distribution and the allele frequency of the C825T polymorphism were investigated in a series of 104 patients with oncocytic thyroid tumours of follicular cell origin [58 adenomas, 41 follicular thyroid carcinomas (FTCs), and five papillary thyroid carcinomas (PTCs)]; the results were compared with those obtained from 321 age and gender matched healthy blood donors and a series of 327 non-oncocytic thyroid tumours of follicular cell origin (119 adenomas, 80 FTCs, and 186 PTCs). Analysis of the genotype distribution (comparing oncocytic with non-oncocytic tumours of the present series) revealed a significantly increased odds ratio (OR) for CC versus TT (OR = 4.22; p = 0.011) and CC versus CT (OR = 1.62; p = 0.049) carriers to develop an oncocytic thyroid tumour; ORs to develop an oncocytic thyroid tumour were also increased comparing the genotype distribution between the group of oncocytic tumours and healthy controls for CC versus TT (OR = 3.73; p = 0.017) and CC versus all T carriers (OR = 1.56; p = 0.034). Oncocytic thyroid tumours as a group showed a statistically significant increase of the C-allele frequency when compared with all non-oncocytic tumours (p = 0.0039) as well as healthy blood donors (p = 0.017). The results strongly suggest that the C allele of the GNB3 C825T polymorphism of the G protein beta3-subunit is associated with an increased risk for the development of oncocytic thyroid tumours. This polymorphism may thus be considered a (co)factor favouring the development of oncocytic thyroid tumours, although the biological mechanism(s) underlying this association remain obscure. PMID- 17136760 TI - Effect of characteristics of compounds on maintenance of an amorphous state in solid dispersion with crospovidone. AB - Solid dispersion (SD) of indomethacin with crospovidone (CrosPVP) shows useful characteristics for preparation of dosage forms. This study aimed to determine the types of drugs that could adopt a stable amorphous form in SD. Twenty compounds with various melting points (70-218 degrees C), molecular weights (135 504) and functional groups (amide, amino, carbonyl, hydroxyl, ketone etc.) were prepared in SD with CrosPVP. The CrosPVP SDs were prepared using a mechanical mixing and heating method. Melting point and molecular weight were found to have no influence on the ability of a compound to maintain an amorphous state in SD. All compounds containing hydrogen-bond-donor functional groups existed in an amorphous state in SD for at least 6 months. Infrared spectra suggested an interaction between the functional groups of these compounds and amide carbonyl group of CrosPVP. Compounds without hydrogen-bond-donor groups could not maintain an amorphous state and underwent recrystallization within 1 month. It was suggested that the presence of a hydrogen-bond-donor functional group in a compound is an important factor affecting the stable formation of SD with CrosPVP, which contains a hydrogen-bond acceptor. PMID- 17136761 TI - Characterization of anhydrous and hydrated pharmaceutical materials with THz time domain spectroscopy. AB - The identification of anhydrous and hydrated forms of pharmaceutical substances is of great importance in pharmaceutical science and industry. We report the use of THz time-domain spectroscopy (THz-TDS) for pseudopolymorph investigation. The anhydrous forms of pharmaceutical substances including caffeine, theophylline, D glucose, and ampicillin exhibit different THz absorption spectra from their hydrated forms, presumably due to their different intermolecular vibrational modes mediated by hydrogen-bonding. The data from X-ray powder diffractometry (XRPD) confirm the crystallinity differences between the anhydrous and hydrated forms. The temperature-dependent THz spectra of caffeine hydrate were also recorded, and it demonstrates that THz-TDS can be used to monitor the dehydration process of drug hydrates. We conclude that THz-TDS is an advantageous technique for the pseudopolymorph identification and study, and has great potential to become a process analytical technology (PAT) in pharmaceutical production and quality control. PMID- 17136762 TI - 5alpha-Reductase type 2 gene variant associations with prostate cancer risk, circulating hormone levels and androgenetic alopecia. AB - Controversy exists over the significance of associations between the SRD5A2 (5alpha-reductase type 2) polymorphisms, A49T and V89L, and risk of prostate cancer. These potentially functional polymorphisms may alter life-long exposure to androgens with subsequent effects on male health and aging. The aim of this study was to examine the association of these variants with prostate cancer risk, plasma hormone levels and androgenetic alopecia. Subjects include 827 cases and 736 controls from an Australian population-based case-control study of prostate cancer. Information on prostate cancer risk factors and patterns of balding were collected. Plasma levels of testosterone, 3alpha-diol glucuronide (3alpha-diolG), dehydroepiandrosterone sulfate, androstenedione, sex hormone-binding globulin and estradiol were measured for controls. No associations with the V89L polymorphism were found. Carriers of the rarer A49T A allele were at a 60% higher risk of prostate cancer (OR = 1.60; 95% CI 1.09-2.36; p = 0.02) and 50% lower risk of vertex and frontal balding (p = 0.03) compared with men homozygous for the more common G allele. Although we found little evidence of association between this variant and plasma levels of 5 measured androgens, circulating 3alpha-diolG levels were 34% lower in A49T A allele carriers (p < 0.0001). Our study provides evidence that the SRD5A2 A49T A variant is associated with an increased risk of prostate cancer, lower levels of circulating 3alpha-diolG and decreased risk of baldness. These findings raise important questions with respect to previous assumptions concerning hormonal influences on prostate cancer risk in ageing males. PMID- 17136763 TI - Mass spectrometric characterization of low-molecular-mass color pI markers and their use for direct determination of pI value of proteins. AB - The use of low-molecular-mass color pI markers for the determination of pI values of proteins in gel isoelectric focusing (IEF) in combination with mass spectrometry is described. Different types of substituted phenols of known pI values within the mass range 250-400 were used here as pI markers. The pure, synthesized pI markers were studied by MALDI-TOF/TOF MS. Fragmentation studies of the pI markers were also performed. Only stable and well-characterized pI markers were used in this work. The selected pI markers were mixed with proteins, deposited on a gel and separated in a pH gradient. Color pI markers enable supervision of progress of the focusing process and also estimation of the position of the invisible focused bands. The separated bands of the pI markers (containing separated proteins) were excised, and the pI markers were eluted from each gel piece by water/ethanol and identified by MALDI-TOF/TOF MS. From the washed gel pieces the remaining carrier ampholytes were then washed out and proteins were in-gel digested with trypsin. The obtained peptides were measured by MALDI-TOF/TOF MS and the proteins identified via a protein database search. This procedure allows avoiding time-consuming protein staining and destaining procedures, which shortens the analysis time roughly by half. For comparison, IEF gels were stained with Coomassie Brilliant Blue R 250 and proteins in the gel bands were identified according to the standard proteomic protocol. This work has confirmed that our approach can give information about the correct pI values of particular proteins and shorten significantly the time of analysis. PMID- 17136764 TI - A study of the glutathione metaboloma peptides by energy-resolved mass spectrometry as a tool to investigate into the interference of toxic heavy metals with their metabolic processes. AB - To better understand the fragmentation processes of the metal-biothiol conjugates and their possible significance in biological terms, an energy-resolved mass spectrometric study of the glutathione conjugates of heavy metals, of several thiols and disulfides of the glutathione metaboloma has been carried out. The main fragmentation process of gamma-glutamyl compounds, whether in the thiol, disulfide, thioether or metal-bis-thiolate form, is the loss of the gamma glutamyl residue, a process which ERMS data showed to be hardly influenced by the sulfur substitution. However, loss of the gamma-glutamyl residue from the mono-S glutathionyl-mercury (II) cation is a much more energetic process, possibly pointing at a strong coordination of the carboxylic group to the metal. Moreover, loss of neutral mercury from ions containing the gamma-glutamyl residue to yield a sulfenium cation was a much more energetic process than those not containing them, suggesting that the redox potential of the thiol/disulfide system plays a role in the formal reduction of the mercury dication in the gas phase. Occurrence of complementary sulfenium and protonated thiol fragments in the spectra of protonated disulfides of the glutathione metaboloma mirrors the thiol/disulfide redox process of biological importance. The intensity ratio of the fragments is proportional to the reduction potential in solution of the corresponding redox pairs. This finding has allowed the calculation of the previously unreported reduction potentials for the disulfide/thiol pair of cysteinylglycine, thereby confirming the decomposition scheme of bis- and mono-S-glutathionyl-mercury (II) ions. Finally, on the sole basis of the mass spectrometric fragmentation of the glutathione-mercury conjugates, and supported by independent literature evidence, an unprecedented mechanism for mercury ion-induced cellular oxidative stress could be proposed, based on the depletion of the glutathione pool by a catalytic mechanism acting on the metal (II)-thiol conjugates and involving as a necessary step the enzymatic removal of the glutamic acid residue to yield a mercury (II) cysteinyl-glycine conjugate capable of regenerating neutral mercury through the oxidation of glutathione thiols to the corresponding disulfides. PMID- 17136765 TI - Methods validation for the determination of trihalomethanes in drinking water. AB - Trihalomethanes (THMs), chlorinated by-products in drinking water, have been determined by comparing some analytical methods, based on the following techniques: liquid-liquid extraction-gas chromatography-mass spectrometry (LLE-GC MS), headspace-gas chromatography-mass spectrometry (headspace-GC-MS) and purge and trap-gas chromatography-mass spectrometry (PT-GC-MS). The mass spectrometer was operated in the SIM mode. The quantitative methods were validated and compared to the ability to identify and to measure reliably the yields of the toxic compounds. Good validation parameters were obtained for each method. PMID- 17136766 TI - Impact of the National Breast and Cervical Cancer Early Detection Program on mammography and Pap test utilization among white, Hispanic, and African American women: 1996-2000. AB - Prevention, including routine cancer screening, is key to meeting national goals for the elimination of death and suffering due to cancer. Since 1991, the U.S. government has invested in programs such as the National Breast and Cervical Cancer Early Detection Program (NBCCEDP) to detect breast and cervical cancer early among uninsured low-income women. A concomitant goal is reducing racial disparities in screening and early detection, and the NBCCEDP program targets low income women who are more often racial and ethnic minorities. This paper analyzes data to test for effects of the NBCCEDP and other determinants of screening across racial/ethnic groups. We used data from the Behavioral Risk Factor Surveillance System (BRFSS) for 1996 through 2000. These data indicate that gaps in testing for breast and cervical cancers between African American and non Hispanic white women aged 40-64 years have closed but remain for Hispanics. Multivariate findings indicate that the longevity of free screening sites through the NBCCEDP significantly increased both tests for non-Hispanic white women. The data do not confirm this effect for other racial and ethnic groups. Analysis did indicate that public insurance, or Medicaid, was equal to private insurance in promoting increased testing for African Americans and Hispanics, but not for non Hispanic whites. Assuring that Medicaid remains available for women in this nonelderly group and increasing access to free screening sites can lead us closer to national screening goals, yet policies still need to address racial/ethnic disparities in insurance and service delivery. Cancer 2007. (c) 2006 American Cancer Society. PMID- 17136768 TI - Current literature in mass spectrometry. PMID- 17136767 TI - Mass spectrometric studies on 4-aryl-1-cyclopropyl-1,2-dihydropyridinyl derivatives: an examination of a novel fragmentation pathway. AB - Examination of the electrospray ionization product ion spectra of 1,2 dihydropyridinyl and 4-aryl-1,2-dihydropyridinyl derivatives bearing a 1 cyclopropyl or 1-trans-2-phenylcyclopropyl group has led to the characterization of unexpected fragment ions. For example, the base peak at m/z 156 present in the product ion spectrum of trans-1-(2-phenylcyclopropyl)-4-phenyl-1,2 dihydropyridine proved not to be the expected 4-phenylpyridinium species but rather the isomeric 3-phenyl-5-azoniafulvenyl species. The results of studies with a series of structural and isotopically labeled analogs require a novel fragmentation pathway to account for the formation of this and related fragment ions. One possible pathway is based on an initial 1,5-sigmatropic shift of a cyclopropylmethylene hydrogen atom that is accompanied by opening of the cyclopropyl ring. The resulting eniminium intermediates then fragment to yield the 5-azoniafulvenyl species. PMID- 17136769 TI - Reply to "a new device for cricothyroidotomy". PMID- 17136770 TI - ROCK inhibition with Y27632 activates astrocytes and increases their expression of neurite growth-inhibitory chondroitin sulfate proteoglycans. AB - Inhibition of Rho-kinase (ROCK) with Y27632 stimulates sprouting by injured corticospinal tract and dorsal column tract axons, and accelerates functional recovery. However, regeneration of these axons across the glial scar was not observed. Here we examined the effects of Y27632 treatment on chondroitin sulfate proteoglycan (CSPG) expression by astrocytes, which are a key component of the reactive gliosis inhibiting axonal regeneration. In vivo, rats underwent a dorsal column transection and were treated with Y27632 via intrathecal pump infusion. Compared with controls, Y27632-treated injury sites displayed exaggerated upregulation of glial fibrillary acid protein and neurocan immunoreactivity along the lesion edge. In vitro, astrocytes assumed a reactive morphology (stellate shape) and increased their expression of CSPGs after Y27632 treatment. Neurite growth by dissociated cortical neurons decreased when cultured on the extracellular matrix (ECM) derived from Y27632-treated astrocytes. This decrease in neurite growth was reversed with chondroitinase-ABC (ChABC) digestion, indicating that the inhibition was due to CSPG depositions within the ECM. Interestingly, conditioned medium (CM) from untreated astrocytes was inhibitory to neurite growth, which was overcome by ChABC digestion. Such inhibitory activity was not found in the CM of Y27632-treated astrocytes. Taken together, these data support a model where ROCK inhibition by Y27632 modifies astrocytic processing of CSPGs, and increases the presence of CSPGs within the ECM while reduces CSPGs in the CM (cerebrospinal fluid in vivo). This increased expression of inhibitory CSPGs in the ECM of the glial scar may counteract the growth promoting effects of ROCK inhibition on axonal growth cones. PMID- 17136771 TI - Evidence for synaptic stripping by cortical microglia. AB - Recent studies have described significant demyelination and microglial activation in the cerebral cortex of brains from multiple sclerosis patients. To date, however, experimental models of cortical demyelination or cortical inflammation have not been extensively studied. In this report we describe focal cortical inflammation induced by stereotaxic injection of killed bacteria (BCG), followed 1 month later by subcutaneous injection of the same antigen, a protocol that overcomes the immune privilege of the cortex. Intracerebral BCG injection produced focal microglial activation at the injection site (termed acute lesion). Ten days after peripheral challenge (termed immune-mediated lesion), larger areas and higher densities of activated microglia were found near the injection site. In both paradigms, activated microglia and/or their processes closely apposed neuronal perikarya and apical dendrites. In the immune-mediated lesions, approximately 45% of the axosomatic synapses was displaced by activated microglia. Upon activation, therefore, cortical microglial migrate to and strip synapses from neuronal perikarya. Since neuronal pathology was not a feature of either the acute or immune-mediated lesion, synaptic stripping by activated microglia may have neuroprotective consequences. PMID- 17136772 TI - Axon-glia communication evokes calcium signaling in olfactory ensheathing cells of the developing olfactory bulb. AB - Olfactory ensheathing cells (OECs) accompany receptor axons in the olfactory nerve and promote axonal growth into the central nervous system. The mechanisms underlying the communication between axons and OECs, however, have not been studied in detail yet. We investigated the effect of activity-dependent neuronal transmitter release on Ca(2+) signaling of OECs in acute mouse olfactory bulb slices using confocal Ca(2+) imaging. TTX-sensitive axonal activity upon electrical nerve stimulation triggers a rise in cytosolic Ca(2+) in OECs, which can be mimicked by application of DHPG, an agonist of metabotropic glutamate receptors (mGluRs). Both stimulation- and DHPG-induced Ca(2+) transients in OECs were abolished by depletion of intracellular Ca(2+) stores with cyclopiazonic acid (CPA). The mGluR(1)-specific antagonist CPCCOEt completely inhibited DHPG evoked Ca(2+) transients, but reduced stimulation-induced Ca(2+) transients only partly, suggesting the involvement of another neurotransmitter. Application of ATP evoked CPA-sensitive Ca(2+) transients in OECs, which were inhibited by the P2Y(1)-specific antagonist MRS2179. Co-application of CPCCOEt and MRS2179 almost completely blocked the stimulation-induced Ca(2+) transients, indicating that they were mediated by mGluR(1) and P2Y(1) receptors. Our results show that OECs are able to respond to olfactory nerve activity with an increase in cytosolic Ca(2+) due to glutamate and ATP release. PMID- 17136774 TI - Sex-specific QTL effects on variation in paraoxonase 1 (PON1) activity in Mexican Americans. AB - Paraoxonase 1 (PON1), a high-density lipoprotein-associated enzyme known to protect against cellular damage from toxic agents, may also have antioxidant properties. PON1 activity levels have been reported to differ by sex in human and animal studies with females exhibiting higher basal levels. We measured PON1 activity frozen serum for 1,406 individuals in over 40 extended pedigrees from the San Antonio Family Heart Study (SAFHS). We used a maximum likelihood-based, variance decomposition approach implemented in SOLAR to test for genotype-by-sex (G x S) interaction on variation in PON1 activity and to determine if any of the four PON1 quantitative trait loci (QTL) previously reported by us for this population might account for sex differences in PON1 activity levels. The residual additive genetic correlation (rho(G) = 0.82) between males and females is significantly different from 1 (P = 0.009), suggesting that some of the genes that influence PON1 activity act differently in females and males or, possibly, that a different combination of genes influences this trait in each sex. In addition to the QTL at or near the PON structural locus on 7q21-22, three other potential QTLs were evaluated for sex-specific effects: one each on chromosomes 12, 17 and 19. The QTL on chromosome 17 (LOD = 2.32, P = 0.0003; flanked by microsatellite marker loci D17S974 and D17S969) shows a significant (P = 0.005) sex-specific effect on PON1 activity; accounting for 6% of the additive genetic variance in males and 20% in females. This study represents the first formal statistical genetic test for G x S interactions on normal quantitative variation in PON1 activity in humans. PMID- 17136775 TI - Non-parametric bounds on treatment effects with non-compliance by covariate adjustment. AB - Consider a clinical trial where subjects are randomized to two treatment arms but compliance to the assignment is not perfect. Concerning this problem, this paper derives non-parametric bounds on treatment effects by making use of the observed covariates information. The new bounds are narrower and more informative than the existing ones. In addition, a new non-parametric point estimation approach is proposed based on stratified analysis. Furthermore, to examine the accuracy of estimating the proposed bounds, we provide variance estimators for the proposed approach. The results of this paper can yield credible information on treatment effects, which will be useful for medical research and public health policy analysis. PMID- 17136776 TI - Combining estimates from multiple early studies to obtain estimates of response: using shrinkage estimates to obtain estimates of response. AB - Development of anti-cancer therapies usually involve small to moderate size studies to provide initial estimates of response rates before initiating larger studies to better quantify response. These early trials often each contain a single tumor type, possibly using other stratification factors. Response rate for a given tumor type is routinely reported as the percentage of patients meeting a clinical criteria (e.g. tumor shrinkage), without any regard to response in the other studies. These estimates (called maximum likelihood estimates or MLEs) on average approximate the true value, but have variances that are usually large, especially for small to moderate size studies. The approach presented here is offered as a way to improve overall estimation of response rates when several small trials are considered by reducing the total uncertainty.The shrinkage estimators considered here (James-Stein/empirical Bayes and hierarchical Bayes) are alternatives that use information from all studies to provide potentially better estimates for each study. While these estimates introduce a small bias, they have a considerably smaller variance, and thus tend to be better in terms of total mean squared error. These procedures provide a better view of drug performance in that group of tumor types as a whole, as opposed to estimating each response rate individually without consideration of the others. In technical terms, the vector of estimated response rates is nearer the vector of true values, on average, than the vector of the usual unbiased MLEs applied to such trials. PMID- 17136777 TI - Platinum(II) complexes with pi-conjugated, naphthyl-substituted, cyclometalated ligands (RC N N): structures and photo- and electroluminescence. AB - The crystal structures and photophysical properties of mononuclear [(RC N N)PtX](ClO4)n ((RC N N)=3-(6'-(2''-naphthyl)-2'-pyridyl)isoquinolinyl and derivatives; X=Cl, n=0; X=PPh(3) or PCy(3), n=1), dinuclear [(RC N N)2Pt2(mu dppm)](ClO4)2 (dppm=bis(diphenyphosphino)methyl) and trinuclear [(RC N N)3Pt3(mu dpmp)](ClO4)3 (dpmp=bis(diphenylphosphinomethyl)phenylphosphine) complexes are presented. The crystal structures show extensive intra- and/or intermolecular pipi interactions; the two (RC N N) planes of [(RC N N)2Pt2(mu-dppm)](ClO4)2 (R=Ph, 3,5-tBu2Ph or 3,5-(CF3)2Ph) are in a nearly eclipsed configuration with torsion angles close to 0 degrees. [(RC N N)PtCl], [(RC N N)2Pt2(mu dppm)](ClO4)2, and [(RC N N)3Pt3(mu-dpmp)](ClO4)3 are strongly emissive with quantum yields of up to 0.68 in CH2Cl2 or MeCN solution at room temperature. The [(RC N N)PtCl] complexes have a high thermal stability (T(d)=470-549 degrees C). High-performance light-emitting devices containing [(RC N N)PtCl] (R=H or 3,5 tBu2Ph) as a light-emitting material have been fabricated; they have a maximum luminance of 63,000 cd m(-2) and CIE 1931 coordinates at x=0.36, y=0.54. PMID- 17136778 TI - Functionalization of heteropolyanions-osmium and rhenium nitrido derivatives of Keggin- and Dawson-type polyoxotungstates: synthesis, characterization and multinuclear ((183)W, (15)N) NMR, EPR, IR, and UV/Vis fingerprints. AB - Reaction of K(10)[alpha(2)-P(2)W(17)O(61)] or K(10)[alpha(1)-P(2)W(17)O(61)] or [Bu(4)N][OsCl(4)N] in a water/methanol mixture, and subsequent precipitation with (Bu(4)N)Br provided [alpha(2)-P(2)W(17)O(61){Os(VI)N}](7-) and [alpha(1) P(2)W(17)O(61){Os(VI)N}](7-) Dawson structures as tetrabutylammonium salts. Reactions of [(Bu(4)N)(4)][alpha-H(3)PW(11)O(39)] with either [ReCl(3)(N(2)Ph(2))(PPh(3))(2)] or [Bu(4)N][ReCl(4)N] are alternatives to the synthesis of [(Bu(4)N)(4)][alpha-PW(11)O(39){Re(VI)N}]. (183)W and (15)N NMR, EPR, IR, and UV-visible spectroscopies and cyclic voltammetry have been used to characterize these compounds and the corresponding [(Bu(4)N)(4)][alpha PW(11)O(39){Os(VI)N}] Keggin derivative. PMID- 17136779 TI - Self-assembled ionophores from isoguanosine: diffusion NMR spectroscopy clarifies cation's and anion's influence on supramolecular structure. AB - Cation-templated self-assembly of the lipophilic isoguanosine (isoG 1) with different monovalent cations (M(+)=Li(+), Na(+), K(+), NH(4) (+), and Cs(+)) was studied in solvents of different polarity by using diffusion NMR spectroscopy. Previous studies that did not use diffusion NMR techniques concluded that isoG 1 forms both pentamers (isoG 1)(5)M(+) and decamers (isoG 1)(10)M(+) in the presence of alkali-metal cations. The present diffusion NMR studies demonstrate, however, that isoG 1 does not form (isoG 1)(5)M(+) pentamers. In fact, the diffusion NMR data indicates that both doubly charged decamers of formula (isoG 1)(10)2 M(+) and singly charged decamers, (isoG 1)(10)M(+), are formed with lithium, sodium, potassium, and ammonium tetraphenylborate salts (LiB(Ph)(4), KB(Ph)(4), NaB(Ph)(4) and NH(4)B(Ph)(4)), depending on the isoG 1:salt stoichiometry of the solution. In the presence of CsB(Ph)(4), isoG 1 affords only the singly charged decamers (isoG 1)(10)Cs(+). By monitoring the diffusion coefficient of the B(Ph)(4) (-) ion in the different mixtures of solvents, we also concluded that the anion is more strongly associated to the doubly charged decamers (isoG 1)(10)2 M(+) than to the singly charged decamers (isoG 1)(10)M(+). The (isoG 1)(10)2 M(+) species can, however, exist in solution without the mediation of the anion. This last conclusion was supported by the finding that the doubly charged decamers (isoG 1)(10)2 M(+) also prevail in 1:1 CD(3)CN:CDCl(3), a solvent mixture in which the B(Ph)(4) (-) ion does not interact significantly with the self-assembled complex. These diffusion measurements, which have provided new and improved structural information about these decameric isoG 1 assemblies, demonstrate the utility of combining diffusion NMR techniques with conventional NMR methods in seeking to characterize labile, multicomponent, supramolecular systems in solution, especially those with high symmetry. PMID- 17136780 TI - Catalysis of a peptidic micellar assembly covalently immobilized within mesoporous silica channels: importance of amphiphilic spatial design. AB - A mesostructured silica/organic composite 1-MS, constructed from a rodlike micelle of amino acid amphiphile 1 that has a condensable head group and that can be used as a template, was found to be able to catalyze the acetalization of cyclohexanone, in ethanol at 25 degrees C (50% in 12 h), whereas no reaction took place with unfunctionalized mesoporous silica. In sharp contrast, hydrolytic removal of the C16 alkyl tail of immobilized 1 resulted in the complete disappearance of the catalytic activity, which suggests the importance of a hydrophobic inner domain for the admission of cyclohexanone. Unsupported peptide amphiphile 2, under identical conditions to those above, was inefficient for acetalization regardless of the absence (2% in 24 h) or presence of mesoporous silica (7% in 24 h). Reference composite 2-MS, which is a noncovalently immobilized peptidic micelle, was virtually inactive (1% in 24 h). These observations indicate the importance of covalent immobilization of the peptidic rod micelle for catalysis. Mesostructured silicate 3-MS hybridized with a nonpeptidic, ammonium ion amphiphile (3) showed a certain catalytic activity, but the yield (12% in 24 h) of the acetal was much lower than that achieved by using 1-MS as the catalyst. Amorphous silica with immobilized 1 on its surface was much less active than 1-MS for acetalization (5% in 24 h). PMID- 17136781 TI - Effects of 3d-4f magnetic exchange interactions on the dynamics of the magnetization of Dy(III)-M(II)-Dy(III) trinuclear clusters. AB - [{Dy(hfac)(3)}(2){Fe(bpca)(2)}] x CHCl(3) ([Dy(2)Fe]) and [{Dy(hfac)(3)}(2){Ni(bpca)(2)}]CHCl(3) ([Dy(2)Ni]) (in which hfac(-)=1,1,1,5,5,5 hexafluoroacetylacetonate and bpca(-)=bis(2-pyridylcarbonyl)amine anion) were synthesized and characterized. Single-crystal X-ray diffraction shows that [Dy(2)Fe] and [Dy(2)Ni] are linear trinuclear complexes. Static magnetic susceptibility measurements reveal a weak ferromagnetic exchange interaction between Ni(II) and Dy(III) ions in [Dy(2)Ni], whereas the use of the diamagnetic Fe(II) ion leads to the absence of magnetic exchange interaction in [Dy(2)Fe]. Dynamic susceptibility measurements show a thermally activated behavior with the energy barrier of 9.7 and 4.9 K for the [Dy(2)Fe] and [Dy(2)Ni] complexes, respectively. A surprising negative effect of the ferromagnetic exchange interaction has been found and has been attributed to the structural conformation of these trinuclear complexes. PMID- 17136782 TI - Extraction of proteins from biological fluids by use of an ionic liquid/aqueous two-phase system. AB - An ionic liquid/aqueous two-phase system based on the hydrophilic ionic liquid 1 butyl-3-methylimidazolium chloride (BmimCl) and K(2)HPO(4) has been employed for direct extraction of proteins from human body fluids for the first time. Proteins present at low levels were quantitatively extracted into the BmimCl-rich upper phase with a distribution ratio of about 10 between the upper and lower phase and an enrichment factor of 5. Addition of an appropriate amount of K(2)HPO(4) to the separated upper phase results in a further phase separation, giving rise to an improved enrichment factor of 20. FTIR and UV spectroscopy demonstrated that no chemical (bonding) interactions between the ionic liquid and the protein functional groups were identifiable, while no alterations of the natural properties of the proteins were observed. The partitioning of proteins in the two phase system was assumed to have been facilitated by the electrostatic potential difference between the coexisting phases, as well as by salting out effects. The system could be applied successfully for the quantification of proteins in human urine after on-line phase separation in a flow system. The use of an ionic liquid, as a green solvent, offers clear advantages over traditional liquid liquid extractions, in which the use of toxic organic solvents is unavoidable. PMID- 17136783 TI - Hexagonal supramolecular architectures from ferrocenium cations incorporating [Ni(mnt)2]- columns: structures and properties of [alkylferrocene][Ni(mnt)2]- charge-transfer complexes (mnt=maleonitriledithiolate). AB - The crystal architecture, magnetic properties, and thermodynamic properties of [n butylferrocene][Ni(mnt)2] (1), [tert-butylferrocene][Ni(mnt)2] (2), [1,1' diethylferrocene][Ni(mnt)2] (3), and [1,1'-diisopropylferrocene][Ni(mnt)2] (4) were investigated (mnt=maleonitriledithiolate). These complexes exhibit a unique supramolecular structure in which the ferrocenium cations constitute honeycomb like assembled structures surrounding columns of the anions. For 1, the cations form a dimer through a very short intermolecular ferrocene-ferrocene distance of 3.28 A, which mediates an antiferromagnetic interaction with a singlet-triplet energy gap of 5 K. First-order phase transitions occur in 1-3 at 364, 361, and 350 K, respectively, accompanied by thermal hysteresis. PMID- 17136784 TI - Pillars, layers, pores and networks from nickeltripyrrins: a porphyrin fragment as a versatile building block for the construction of supramolecular assemblies. AB - Sterically hindered nickel- tripyrrins [Ni(trpy)X] with different di-, tri- and tetradentate anions X have been prepared with the aim of finding coordination polymers formed by self-association. The syntheses were performed by simple ligand-exchange reactions and proceeded successfully with the pseudohalides CN( ), OCN(-), SCN(-), SeCN(-), N(CN)(2) (-) (dicyanoamido, dca) and C(CN)(3) (-) (tricyanomethanido, tcm), the cyanidometallates [Ag(CN)(2)](-) and [Ni(CN)(4)](2 ) and the salicylate anion (sal(-)). X-ray crystallographic analyses revealed that the complexes with cyanido and isocyanato ligands, as well as the compound with a salicylato ligand, are prototypes for structurally distinct monomeric species in the solid state, whereas one-dimensional coordination polymers or supramolecular three-dimensional networks are formed from all other combinations. The polymeric compounds display a variety of individual pillar and network architectures with functionalised pores and clefts and with the Ni(trpy) fragments in different relative orientations. Hydrogen bonding and pi stacking were found to be additional structure-directing effects, which increased the structural complexity of the system. The Ni(trpy) subunit has thus been proven to be a versatile building block for the construction of supramolecular assemblies and metal organic frameworks (MOFs) from pentacoordinate Ni(II) ions. PMID- 17136785 TI - Electrostatic self-assembly of polystyrene microspheres by using chemically directed contact electrification. PMID- 17136786 TI - Controlled stacking of 10 transition-metal ions inside a DNA duplex. PMID- 17136787 TI - Container molecules with portals: Reversibly switchable cycloalkane complexation. PMID- 17136788 TI - Protein recognition and denaturation by self-assembling fragments on a DNA quadruplex scaffold. PMID- 17136789 TI - Molecular tectonics on surfaces: Bottom-up fabrication of 1D coordination networks that form 1D and 2D arrays on graphite. PMID- 17136790 TI - A general method for the preparation of ethers using water-resistant solid lewis acids. PMID- 17136791 TI - Design of a practical fluorescent probe for superoxide based on protection deprotection chemistry of fluoresceins with benzenesulfonyl protecting groups. AB - A strategy for designing probes based on protection-deprotection chemistry involving fluoresceins and their benzenesulfonyl (BES) derivatives has led to the development of a much more practical superoxide (O(2) (-.)) probe than the previously reported bis(2,4-dinitro-BES) tetrafluorofluorescein (6 a). Examination of various BES derivatives, developed from the starting point of the prototype probe 6 a, yielded 4,5-dimethoxy-2-nitro-BES tetrafluorofluorescein (BESSo; 7 j) as the optimal reagent. A microtiter plate assay with BESSo showed a tenfold improved detection limit for O(2) (-.) compared with such an assay based on 6 a. BESSo showed markedly better specificity for O(2) (-.) than for GSH or other reactive oxygen species, and this specificity was significantly higher than that of Fe(2+) and some reducing enzymes. These features have resulted in the development of an assay based on BESSo that is capable of providing more unambiguous results for O(2) (-.) release from neutrophils, with or without stimulation by phorbol myristate acetate, as compared with an assay based on 6 a. Intracellular generation of O(2) (-.) in human Jurkat T cells stimulated by butyric acid has been measured by using flow cytometry and fluorescence microscopy utilizing the acetoxymethyl derivative of BESSo. PMID- 17136792 TI - Selection and identification of human gonadotropin-releasing hormone promoter binding peptides by phage display-CEMSA. AB - Specific interactions between transcription factors and cis-acting DNA sequences form the molecular basis of gene expression regulation. Here, we applied phage display technology to DNA-protein interaction studies. A phage-displayed peptide library was used to select Gonadotropin-releasing hormone promoter (GP) binding peptides. After four sequential rounds of biopanning on GP-conjugated magnetic beads, phage clones encoding GQPTPRNAGLPL (B6), SRLNVEPLTTYS (B3), and TTLHWASLTTGR (B11) were enriched. Phages bearing these peptides showed specific binding to GP in solution by capillary electrophoresis mobility shift assay (CEMSA). In addition, some human transcription factors were speculated as the potential transcription factors or co-activators of GnRH gene by bioinformatic analysis. These results suggest that phage display-CEMSA methodology should be a powerful tool to screen and identify site-specific DNA-binding peptides. PMID- 17136793 TI - Fit for health. Everyone agrees it's good to be physically fit, but exactly what should you strive for? PMID- 17136794 TI - Polymyalgia rheumatica. Getting out of bed in the morning can be a challenge- especially for the many people who have this inflammatory condition. PMID- 17136795 TI - Recognizing domestic partner abuse. With its daunting complexities, the path to change takes courage and support. PMID- 17136796 TI - By the way, doctor. I'm hearing more and more about deterioration of the jawbone associated with bisphosphonate drugs. I've been taking Fosamax for over six years for osteoporosis. What should I do? PMID- 17136797 TI - Nonalcoholic fatty liver disease. A growing problem. PMID- 17136798 TI - Health tips. Ladder safety. PMID- 17136799 TI - Possible link between high cholesterol and prostate cancer. PMID- 17136800 TI - Arthritis and the big toe. Relief for a common problem. PMID- 17136801 TI - Hearing loss. How loud is too loud? PMID- 17136802 TI - Is walking outside better exercise than walking on a treadmill? PMID- 17136803 TI - The cervical cancer vaccine. A new vaccine promises to save lives, but won't replace the Pap test. PMID- 17136804 TI - Multivitamins: should you buy this insurance? Studies have raised doubts about vitamins, but the multivitamin pill is still a good idea. PMID- 17136805 TI - Bone drugs may harm what they're supposed to protect. The bisphosphonate drugs have been linked to death of bone tissue. PMID- 17136806 TI - By the way, doctor. Is it possible to have one or more TIAs in the past four years and not have them show up in four MRIs? PMID- 17136807 TI - Largely-hidden HIV epidemic among adolescents creates challenge for clinicians. PMID- 17136809 TI - Hurricane Katrina's effect on research at one HIV clinic. PMID- 17136808 TI - HPV vaccination: important for perinatally-infected adolescents. PMID- 17136810 TI - [Regional meeting of the Japanese Circulation Society. 1991. Japan. Abstracts]. PMID- 17136811 TI - Faces of spine care: from advanced imaging. Severe thoracic kyphoscoliosis in a Klippel-Feil patient with complete cervical spine fusion and deformity--primum non nocere. PMID- 17136812 TI - Norris E. Dodd and the connections between domestic and international agricultural policy. PMID- 17136813 TI - Abstracts from the 4th International Conference on Language and Cognitive Development in Down Syndrome, Portsmouth, United Kingdom, 15-18 September 2005. PMID- 17136814 TI - Health, capability, and justice: toward a new paradigm of health ethics, policy and law. PMID- 17136815 TI - Abstracts of the Annual Meeting of the Hungarian Society for Microbiology, October 18-20, 2006, Keszthely, Hungary. PMID- 17136816 TI - Effect of fluoridated public water supplies on dental caries prevalence. 1956. PMID- 17136817 TI - Autophagy as mechanism for cell death in degenerative aortic valve disease. AB - Once degenerative aortic valve disease becomes symptomatic, valve replacement is necessary for prognostic and symptomatic reasons. In elderly patients, symptoms of degenerative aortic valve can often be doubtful. Therefore, it is difficult but important to distinguish patients who need surgery from those who do not. Estimation of the rate of the progression of this disease can be helpful herein because one needs to bear in mind that aortic valve degeneration is an active process, which can influence the rate of progression. Recently, autophagy was discovered as a mechanism of cell death in different cardiovascular diseases such as atherosclerosis, aortic valve degeneration, heart failure and at regions around heart infarctions. Thus understanding autophagy in all its details can be helpful to contribute insights into the cell death machinery of cardiovascular diseases. This could open ways for inhibition of cell death in cardiovascular disease and possibly define targets for future drug design. PMID- 17136818 TI - Novel therapeutic directions for the natriuretic peptides in cardiovascular diseases: what's on the horizon. AB - Remarkable advances have occurred since the discovery of ANP. The field of the natriuretic peptides has moved beyond their role as regulators of renal function. Growing evidence has now established the natriuretic peptides as regulators of myocardial structure and function as well as in cardiovascular regulation. Further, they emerge as being products not only of the cardiomyocyte but also of the cardiac fibroblast. Studies in vitro and in vivo have clearly established these cardiac hormones as antihypertrophic and antifibrotic. Further, indications such as cardioprotection for acute myocardial infarction, chronic therapy with oral BNP for hypertension and next generation peptides such as DNP-like peptides for acute heart failure may be in the near future. Novel therapeutic strategies to be tested in clinical trials in humans with cardiovascular disease are clearly on the horizon. PMID- 17136819 TI - [Prophylactic intraaortic balloon pumping preserves left ventricular systolic function in acute anterior myocardial infarction without cardiogenic shock]. AB - OBJECTIVES: Left ventricular function and prognosis were evaluated in patients with acute myocardial infarction who underwent primary percutaneous coronary intervention supported by intraaortic balloon pumping. METHODS: Fifty-eight consecutive patients with first acute myocardial infarction were treated between July 1999 and April 2006. Twenty-five had cardiogenic shock on admission, whereas 33 did not. Patients with anterior acute myocardial infarction without cardiogenic shock were divided into the prophylactic intraaortic balloon pumping group (Group 1; n=17) and the rescue intraaortic balloon pumping group (Group 2; n=9). RESULTS: Thirty-day in-hospital mortality was 52% in cardiogenic shock patients, and 3% in non-shock patients. Baseline characteristics of non-shock anterior acute myocardial infarction were similar including Thrombolysis in Myocardial Infarction (TIMI) risk scores (5.1 and 5.0) in the two groups. However, average left ventricular ejection fraction in the convalescent stage was superior in Group 1 (48.7% vs. 37.8%, p = 0.03). Thirty-day in-hospital mortality was 0% in Group 1 and 11% in Group 2 (p = 0.34). Cox's hazard ratio in Group 2 to Group 1 was 2.38 (95% confidence intrerval; 0.84-11.1, p = 0.09) in terms of the subsequent major cardiac events. CONCLUSIONS: Prophylactic use of intraaortic balloon pumping starting prior to primary percutaneous coronary intervention preserves the convalescent left ventricular systolic function in patients with high risk for anticipated cardiac events after anterior acute myocardial infarction without cardiogenic shock. PMID- 17136820 TI - [Promising efficacy of the sirolimus-eluting stent in patients with acute myocardial infarction]. AB - OBJECTIVES: This study investigated the safety and efficacy of sirolimus-eluting stents (SESs) on early and late outcomes in patients with acute myocardial infarction. METHODS: A series of 100 consecutive patients (September 2004 to November 2005)with acute myocardial infarction undergoing primary stenting using SES ptember 24 hr) was compared with 100 consecutive patients (September 2003 to August 2004) treated with bare metal stent (BMS). The frequency of major adverse cardiac events (MACE) and stent thrombosis, and status of ticlopidine administration were assessed at 270 days. RESULTS: The rates of premature discontinuation of ticlopidine (SES group <3 months: 11%, BMS group <1 month: 11%, p = NS) and stent thrombosis (SES group: 1%, BMS group: 0%, p = NS) were similar in the two groups. At follow-up, restenosis rate and target vessel revascularization rate were lower in the SES group(4% vs 19%, p < 0.001 and 4% vs 10%, p = 0.149, respectively). Furthermore, the occurrence of MACE at 270 days was significantly less frequent in the SES group compared with the BMS group (6% vs. 17%, p = 0.038). Multivariate analysis showed SES use tended to predict 270 day MACE (hazard ratio 0.37, 95% confidence interval 0.14-1.02, p = 0.055). Culprit lesion located in the left main trunk was identified as an independent predictor of 270-day MACE (hazard ratio 5.43, 95% confidence interval 1.07-27.59, p = 0.041). CONCLUSIONS: The use of a SES was not associated with increased risk of stent thrombosis compared with a BMS. With lower rates of restenosis and subsequent target vessel revascularization, SES placement could provide superior outcomes in patients with acute myocardial infarction. PMID- 17136821 TI - [Assessment of left ventricular diastolic function with pioglitazone in type 2 diabetic patients]. AB - OBJECTIVES: This study investigated the effect of pioglitazone, an insulin sensitizer, on left ventricular diastolic function in type 2 diabetic patients. METHODS: Various echocardiographic parameters such as left ventricular ejection fraction, transmitral E/A ratio (E/A), deceleration time of E, isovolumetric relaxation time and left ventricular Tei index were measured. A total of 59 patients were enrolled in the ON-group, and the parameters were examined before and after pioglitazone administration (15-30 mg/day) for 6 months, and 9 patients in the OFF-group, and the parameters were examined before and 6 months after cessation of pioglitazone. RESULTS: Average age of subjects was 61.5 years with an age range from 45 to 79 years. Left ventricular ejection fraction and brain natriuretic peptide were not significantly changed both in the ON-group and in the OFF-group. Pioglitazone significantly increased E/A (0.91 +/- 0.23 vs 0.99 +/ 0.24, p < 0.01)and decreased deceleration time of E (233.9 +/- 58.6 vs 209.9 +/- 38.3 msec, p < 0.01), isovolumetric relaxation time(91.5 +/- 13.6 vs 76.0 +/- 12.1 msec, p < 0.01)and left ventricular Tei index (0.38 +/- 0.11 vs 0.35 +/- 0.09, p < 0.01) in the ON-group. On the other hand, E/A (1.08 +/- 0.35 vs 0.88 +/ 0.20, p < 0.01) was significantly decreased and left ventricular Tei index (0.28 +/- 0.11 vs 0.33 +/- 0.11, p < 0.01) was significantly increased in the OFF group. CONCLUSIONS: Pioglitazone administration improves and cessation worsens left ventricular diastolic function. PMID- 17136822 TI - Postprandial hyperglycemia is a possible contributor to paroxysmal atrial fibrillation: a case report. AB - Atrial fibrillation, a major risk factor for stroke, is believed to occur first as paroxysmal episodes, gradually becoming more persistent, and finally progressing to chronic atrial fibrillation. Treatment of paroxysmal atrial fibrillation is an important target to prevent chronic atrial fibrillation. We describe a very unique case with postprandial hyperglycemia and obesity associated with drug-refractory paroxysmal atrial fibrillation. A 73-year-old Japanese woman with postprandial hyperglycemia suffered from drug-refractory paroxysmal atrial fibrillation. A 1600 kcal/day diet and walking three times/day for more than 30 min eliminated paroxysmal atrial fibrillation after 6 months. Diet and exercise should be considered as the initial therapy in patients with paroxysmal atrial fibrillation who also have postprandial hyperglycemia. This case suggests that postprandial hyperglycemia and insulin resistance might be one of the possible underlying mechanisms of paroxysmal atrial fibrillation. PMID- 17136823 TI - [Ampulla cardiomyopathy induced by meningitis: two case reports]. AB - Two patients presented with ampulla cardiomyopathy induced by meningitis. A 71 year-old man with meningitis was admitted to our neurosurgery division. Emergent coronary angiography was performed, because of sudden blood pressure fall and ST elevation in the precordial leads. Left ventriculography and coronary angiography revealed apical ballooning without coronary stenosis. A 73-year-old woman with meningitis was admitted to another hospital. She felt chest pain. Electrocardiography showed ST elevation in the precordial leads. She was transferred to our division. Echocardiography revealed apical ballooning and hyperkinesis of the base. Creatine kinase level showed no elevation on admission or 8 hr later. Ampulla cardiomyopathy with cerebrovascular disease is common, but rare with meningitis, which needs intensive care because of the risk of respiratory arrest. PMID- 17136824 TI - [Differentiation of acute coronary syndrome and ulcerative colitis as causes of elevated macro creatine kinase in a patient with effort angina pectoris: a case report]. AB - A 65-year-old man had been medically treated under a diagnosis of ulcerative colitis for 12 years as an outpatient at another facility. Two weeks before admission, he complained for the first time of mild chest pain limited to exertion. He was seen again by his previous physician in the morning and admitted to that facility later that day. Laboratory findings showed elevated serum creatine kinase (CK) at that time, so non-ST-elevated acute coronary syndrome was suspected and the patient was referred to our hospital. On admission, CK-MB was elevated but troponin T was negative. No findings of myocardial damage were observed during the admission period. Elective coronary angiography was performed and the diagnosis was effort angina pectoris. Elevated CK activity persisted but was not cardiogenic and instead due to the presence of elevated macro CK-1 associated with ulcerative colitis. Troponin T and isoenzyme electrophoresis were highly specific for the differential diagnosis of non-cardiogenic elevated CK. PMID- 17136825 TI - [Dipyridamole-induced regular R-R intervals in a case of old myocardial infarction with atrial fibrillation-flutter]. PMID- 17136826 TI - [Sjogren's syndrome]. PMID- 17136827 TI - [Highlights in gastroenterology 2006]. PMID- 17136828 TI - [Negligent anamnesis]. PMID- 17136829 TI - [Resistant anemia in resistant patients]. AB - A 93 year old man with a severe anaemia with possible origin in the gastrointestinal tract refused any clarifying examinations except palliative blood transfusions. Within half a year, he received 48 red blood cell transfusions without any diagnostic or other therapeutically steps. To further decide on treatment strategies, we discussed this problematic case with our local ethic committee and found a reasonable treatment solution PMID- 17136830 TI - [Roentgen diagnostics with the emphasis on it's traditional roentgenological method is a major sourse to improve gastric cancer diagnosis]. AB - The main idea of the article is presentation of todays problem of gastric cancer diagnosis. Authors, supported by long-term experience of gastric cancer diagnostics, demonstrate the necessity of the return of traditional roentgenological method for it's diagnostics, along with endoscopic technique. In the first place it is determined by the increase of sclerotic and mixed forms. In the second place, it is associated by the change of gastric cancer primary localizations, supporting the need of active use of roentgen diagnostics for it's diagnosis. One of the important parts of the article consists of the author's attempt to change general opinion about insignificancy role of traditional roentgenological method for diagnosis of gastric cancer. Authors demonstrate modern techniques of traditional stomach roentgenology and firstly, it's digital development with CR-systems for the improvement of diagnosis of the intramural forms of carcinomas. In the initial stages it is obligatory to engage classical roentgenology with double contrast study and endoscopy as main equally important methods. The unfavourable situaition in the diagnosis of gastric cancer can be changed only with conjoint use of these two approaches. It is chiefly depends with the difficulties of endoscopic study of diffuse (endophytic) forms of gastric carcinoma. Authors separately underline the necessity of including specialists in roentgen diagnostics in the national project of modernization of roentgenological equipment in medical institutions. PMID- 17136831 TI - [Matters of roentgen diagnostics of idiopathic interstitial pneumonia]. AB - This article is devoted to roentgen diagnostics of idiopathic interstitial pneumonia. 7 types of idiopathic interstitial pneumonia are described, including clinical, morphological and roentgenological characteristics. X-ray methods are presented as possible techniques for determination of the type of the idiopathic interstitial pneumonia. PMID- 17136832 TI - [Lung perfusion scanning in the comprehensive diagnosis of pulmonary thromboembolism]. AB - The aim of the study was to define the sensitivity, specificity, and diagnostic accuracy of lung perfusion scanning (LPS) in pulmonary thromboembolism (PTE). PTE diagnostic techniques are comparatively assessed. The data on 108 patients with suspected PTE and lung perfusion defects revealed at pulmonary scintigraphy were analyzed. The diagnostic techniques included electrocardiography (ECG), 150 echocardiography, venous ultrasonography, chest X-ray, and LPS. The significant signs of PTE were singled out of 150 ones (history data, complaints, clinical symptoms, instrumental findings, autopsy data); LPS data were analyzed in detail. The sensitivity, specificity, and accuracy of LPS were 95.2, 20, and 77.7%, respectively. It is shown that lung scans should be interpreted, by taking into account X-ray data, and LPS should follow ECG, venous ultrasonography, and chest X-ray. PMID- 17136833 TI - [Role of spiral computed tomographic angiography in the diagnosis of aortic aneurysmal complications]. AB - The paper considers the basic diagnostic capacities of computed tomographic angiography (CTA) in aortic aneurysmal complications. The results of emergency of CTA were analyzed In 63 patients with suspected aortic aneurysmal dissection and rupture. CTA provides complete information on the site, extent, and spread, form and sizes of an aneurismal process, on the presence of intraluminal thrombi, on the presence or absence of aortic wall dissection and rupture, on the involvement of aortic branches, and the state of adjacent tissues and organs. PMID- 17136834 TI - [Clinical and roentgenological characteristics of synostosis frontal plagiocephaly, induced by early synostosis of cranio-cerebral sutures]. AB - In this study authors demonstrate description and analysis of clinical and roentgenological presentations of early synostosis of cranio-cerebral sutures in children. Roentgenological description of cranio-cerebral sutures in infants showed up their significant narrowing and the increase in shadow density, early appearance of ossifications zones in the peripheral parts of the sutures, diffuse increase of cranial bones thickness because of apposition osteogenesis disorder. Results of the study show that many alterations in the cerebral cranium are accompanied with malformations of facial skeleton. PMID- 17136835 TI - [X-ray morphometric evaluation of the location of an intercalated fragment in double fractures of the shoulder]. AB - The Academician G.A. Ilizarov Russian Research Center "Reconstructive Traumatology and Orthopedics" treated 40 patients with double humeral fractures. On examining their X-ray films, the patients were divided into groups according to the location of an intercalated fragment. The Eletto procedure was used to estimate the levels of fractures. Comparison of the X-ray morphometric studies, the diagram of the humeral architectonics, and clinical observations revealed a relationship between the time of consolidation and the location of an intercalated fragment, which is explained by the degree of preservation of the latter's intramedullary blood flow. While treating patients with double fragments of the shoulder, it is expedient to use the Eletto index to determine the levels of the fragments. PMID- 17136836 TI - [Successful renal angioplasty after postsurgical stenosis of renal artery (clinical case)]. PMID- 17136837 TI - [Pharmacokinetic essentials of adverse reactions of radiopaque contrast agents]. PMID- 17136838 TI - [Preclinical diagnosis of cardiovascular diseases in workers engaged into industries with radiation danger]. AB - The authors presented results of single-stage cross study of randomized select from a cohort of long seniority workers of isotope separation plant in Siberian Chemical Enterprise, with studies of inflammation markers, vascular and platelet hemostasis, serum lipid parameters, calculated atherogeneity coefficient, homocysteine levels. Serum atherogeneity coefficient and inflammation markers levels (fibrinogen, C-reactive protein) could be considered as the most informative laboratory indicators of underlying cardiovascular diseases in individuals without ischemic heart disease. Among a list of standard inductors of platelet aggregation, collagen and epinephrine should be considered preferential. Increased platelet aggregation requires disaggregant therapy, especially in smokers. Increased homocysteine level is associated with high frequency of atherosclerotic plaques and smoking. PMID- 17136839 TI - [State of broncho-pulmonary system in liquidators of Chernobyl power accident consequences]. AB - Liquidators of Chernobyl power accident consiquences were subjected to simultaneous external gamma-beta radiation and inhalation of particles containing radionuclides aerosols, nonradiation toxic chemicals and dust. Typical defects of medical examination of liquidators were seen: sending people with pulmonary diseases to post-accident work, erroneous interpretation of the disease severity with underestimation of previous occupational history and nonradiation factors deteriorating the case. PMID- 17136840 TI - [Micronuclear test evaluating cytogenetic effects of occupational radiation in low doses]. AB - The authors compared cytogenetic injuries level among "Radon" staffers subjected and not subjected to occupational radiation. Both groups compared demonstrated no reliable differences in spontaneous frequency of micronuclear cells. Micronuclear analysis showed no dependence of the injuried cells level on duration of work with radioactive substances and on doses of occupational radiation. Finding was significant increase in lymphocytes radiosensitivity in individuals subjected to occupational radiation. This group appeared to contain the staffers with micronuclear cells share more than 2 times exceeding average values. PMID- 17136841 TI - [Automated system of radiation monitoring in Moscow region]. AB - The authors consider one trend in activities of "Radon"--Center of Radiation and Ecologic Control. The article covers foundation reasons and longstanding function results of operative radiation monitoring system for large industrial city. Features of construction and functioning of the system are shown. Devices for operative radiation monitoring are subjected to comparative analysis. Prospective trends of the system development are described. PMID- 17136842 TI - [Studying patterns of distribution of 137Cs and natural radionuclides in bottom sediments of water ecosystem in Moscow region]. AB - The authors evaluated distribution of technogenic isotope cesium-137 and natural radionuclides (radium-226, thorium-232 and kalium-40) in bottom sediments of water ecosystem in Moscow region. Radiation parameters of the sediments appeared to be similar to background values of soils and grounds in the region. PMID- 17136843 TI - [Cytogenetic control in exposure to ionizing radiation sources (review of literature)]. PMID- 17136844 TI - [Characteristics of utilization object--nomenclature of ampoules with sources of high residual activity gamma-radiation]. AB - The article is a first step in creation of general catalogue for ampoules with sources of high (over 1 x 10(7) Bq) activity gamma-radiation. PMID- 17136845 TI - [Tritium in "radon" special industrial complex, contents of environment]. PMID- 17136846 TI - [Correction of NO-dependent pulmonary vasodilation disorder in cardiosurgical patients with acute respiratory distress syndrome]. AB - Disturbances in endogenous NO synthesis present an important pathogenetic mechanism of NO-dependent vasodilation disorder in patients with acute respiratory distress syndrome. Inhalational NO is an effective means of correcting NO-dependent pulmonary vasodilation and the pulmonary function of oxygenation in 47% of patients with acute respiratory distress syndrome. PMID- 17136847 TI - [Vasculopathy of the grafted heart: the synergism of proinflammatory and proatherogenous factors and viral infection]. AB - The authors analyze the pathogenetic significance of hyperhomocysteinemia, antiphospholipid syndrome, the hyperexpression of cell adhesion molecules, inflammation, and oxidative disorders for, as well as the role of viral infections in the development of coronary artery disease of the grafted heart. The paper shows that viral infections in recipients lead to the development of proinflammatory, proatherogenous, and prothrombogenous status, expressing themselves in an increase in the corresponding laboratory markers in recipients' blood plasma, and points out the role of viral infection in the pathogenesis of coronary artery disease of the transplanted heart. Control and treatment of viral infections, as well as pharmacocorrection of proinflammatory, proatherogenous, and prothrombogenous status would made it possible to influence the development of coronary artery disease of the grafted heart. PMID- 17136848 TI - [Inflammation and apoptosis in relation with the effectiveness of bone marrow cell transplantation to patients with chronic heart failure]. AB - The authors analyze the pathogenetic significance of inflammation, immune activation, and apoptosis in the development of heart failure and the functional regeneration of left ventricular myocardium after treatment including autologic bone marrow cell transplantation. The paper shows that autologic bone marrow cell transplantation influences the same mechanisms that cause heart failure, namely inflammation, apoptosis, and neurohumoral mechanisms. Investigations have made it possible to formulate guidelines for the prediction and monitoring of the effects of treatment including autologic bone marrow cell transplantation in patients with heart failure. PMID- 17136849 TI - [The peculiarities of bone exchange disorder and its regulation in men after long terms following orthotopic heart transplantation and cadaveric kidney transplantation]. AB - The study found bone exchange disorder manifested by accelerated bone resorption, retarded bone formation, and the loss of the bone mineral density (BMD) of the axial and peripheral skeleton in 19 men (39 observations) 66 +/- 44 months following orthotopic heart transplantation (OTHT) and in 92 men 45 +/- 28 months after cadaveric kidney transplantation. An accelerated bone resorption, more pronounced in cadaveric kidney (CK) recipients, is associated with hyperparathyroidism (HPT) and renal dysfunction, while bone formation retardation is associated with a decrease in insulin-like growth factor-1 level. An increase in osteoprotegerin level is of compensatory character. The prominence of HPT depends on the degree of renal dysfunction; in CK recipients it also depends on the degree of the reduction in the levels of biologically active testosterone and estradiol. Reduction in BMD of the peripheral skeleton after OTHT are associated with the degree of renal dysfunction and a decrease in free testosterone index; after CK transplantation it is associated with HPT, the cumulative dose of glucocorticoids, reduction in the levels of biologically active testosterone and estradiol, as well as sex-hormone binding globulin (SHBG); reduction in spine BMD is only associated with SHBG. PMID- 17136850 TI - [Auxiliary blood circulation and the artificial heart: forty years of development]. AB - The article summarizes forty-year experience in working over the problem of the artificial heart and auxiliary blood circulation in Laboratory and later Research Institute of Transplantology and Artificial Organs. This work has resulted in the development of balloon pumps for intraaortic contrapulsation, the technique of balloon installation, and indications to its application. The results of the clinical application of the method in patients with various pathologies have been estimated. Preclinically, left ventricular bypass techniques have been tested on more than twenty models of membrane type artificial ventricles. The results of the clinical application of left ventricular bypass have been analyzed. The authors adduce data on the development of the thermomechanical implantable bypass system "Micron-M". After achieving 100-day survival of calves with an artificial heart, an artificial heart with external power supply was used as a "bridge" to heart transplantation, but there was only one patient in whom transplantation was performed. The authors consider development of implantable auxiliary blood circulation devices and the artificial heart to be of great prospective value. PMID- 17136851 TI - [Coronary artery disease after renal transplantation: epidemiology, risk factors, and surgical approaches to treatment]. AB - Coronary artery disease (CAD) is the main cause of death in renal transplant recipients. The aim of the present study was to determine the frequency and risk factors of post-transplantation CAD and its influence on the long-term results of surgery, as well as to evaluate the efficiency of myocardial revascularization in patients with severe CAD. Analysis of the observation of 479 renal recipients (332 men and 147 women) aged 38.69 +/- 11.2 was performed. The mean follow-up period was 64.56 +/- 37.44 months. Sixty-eight patients had diabetes mellitus. CAD was diagnosed in 14.8% (71 out of 479) renal recipients; in 12.7% of patients it developed de novo and was revealed 32.4 +/- 18.6 months after the surgery. Ten year survival of renal recipients with CAD was only 39%, while in the group of non-CAD patients it was 75% (p < 0.0001). Age more than 45, male gender, diabetes mellitus, hypercholesterolemia, infections, pre-existing left ventricular myocardial hypertrophy, and renal transplant dysfunction were defined as significant risk factors of CAD de novo. Multi-factor Cox model found only age more than 45 (p < 0.009), male gender (p < 0.00001), and hyperlipidemia (p < 0.0058) to be independent risk factors of CAD. Myocardial revascularization was performed in 29 patients with coronary lesions: 27 patients underwent percutaneous transluminal coronary angioplasty with stenting and 2 patients underwent coronary artery bypass grafting (5 and 52 months after renal transplantation). However, angioplasty had to be repeated in 6 out of 27 (22%) patients within 3 to 6 months. The average follow-up duration was 23 months (2 to 74 months) after revascularization. Prolonged effect (more than 12 months) was achieved in 17 out of 29 (58.6%) patients. None of the patients developed myocardial infarction after revascularization. Two patients died 28 and 35 months after angioplasty due to extracardial complications (hepatic cirrhosis and an oncological disease); one patient died 78 months after repeated revascularization from progressive cardiac insufficiency while receiving dialysis due to a relapse of renal transplant insufficiency. Thus, CAD develops in 14.8% of renal transplant recipients; in 12.7 of patients it develops de novo. There are conventional and nonconventional post-transplantation CAD risk factors, which include renal transplant dysfunction and post-transplantation infections. Association with myocardial hypertrophy, observed in a significant number of patients, is a feature of post-transplantation CAD. Coronary revascularization, angioplasty with stenting in particular, may be considered to be an effective method of CAD treatment in renal transplant recipients. PMID- 17136852 TI - [The modern potential of histocompatibility in clinical kidney transplantation]. AB - To many clinicians, the improvement of cadaveric kidney graft survival, achieved in the last decade, has become a reason to reconsider the importance of HLA matching in renal transplantation under modem conditions. Nevertheless, there still are arguments to continue selecting donors on the basis of HLA compatibility. Single-center cadaveric renal transplantation data were used to calculate graft survival rates by Kaplan-Meyer analysis and to create the Cox model of the probability of seven-year transplanted kidney survival. One-year and seven-year survival rates for 362 cadaveric kidneys transplanted in 1996-2004 were 83.1% and 60.7%, respectively. One-factor analysis found five factors--HLA DR compatibility, common HLA epitope compatibility, HLA-DQl compatibility, the replacement of Azathyoprine with Cellsept, and donor's age--to have significant effect on transplant survival rate. All these factors were used as variables in a highly significant (p < 0.00001) multifactor Cox model of graft survival. The summary influence of histocompatibility factors on renal graft survival was found to be as high as 86% compared to 6% related to Cellsept and 8% related to donor's age. These results show that histocompatibility is still the main factor favoring the survival of cadaveric renal allograft. PMID- 17136853 TI - [Hyperhydration and the cardiovascular system in patients receiving hemodialysis]. AB - In order to evaluate the influence of volume overload on the cardiovascular system, patients on program hemodialysis were examined using ultrasonography and isotope ventriculography. The study revealed a doubtless correlation between hyperhydration, on the one hand, and arterial hypertension, an increased minute volume, and left ventricular hypertrophy, on the other. A growing left atrial contribution was shown to play an important role for the maintenance of an adequate left ventricular filling during a session of hemodialysis with ultrafiltration. The study also found paradoxal blood sequestration in the venous system during the process of ultrafiltration in patients with high parathyroid hormone levels. PMID- 17136854 TI - [Cytomegalovirus infection of allografted kidneys in children]. AB - The study was based on 90 puncture bioptats of transplanted kidneys, received from 67 patients (33 males and 34 females) aged 8 to 18 or 14.1 +/- 4.7 years. The puncture biopsies were performed 8 days to 7 years after renal transplantation. In addition to histological techniques, the researchers used immunoperoxidase method with monoclonal antibodies to CMV-E and CMV-L antigens. The immunohistochemical method of CMV infection diagnostics revealed CMV infection in 15 (17.5%) of the bioptats of the renal allografts. The risk of CMV infection in renal transplants increases during the late postoperative period in the presence of chronic pathology (chronic nephropathy or chronic glomerulonephritis), especially in combination with late acute allograft rejection crises. PMID- 17136855 TI - [Chronic hepatitis B and C in renal graft recipients]. AB - Clinical and morphological features of chronic hepatitis B (CHB), C (CHC), and B+C (CHB+C) were studied in 283 renal graft recipients. High total bilirubin serum levels were detected significantly more often in CHB and CHB+C patients vs. CHC patients. High ALT activity was noted in 65% of CHB patients and only in 45% of CHC patients (p = 0.003). Stable low activity of hepatitis prevailed in renal recipients; it was noted in 56.7% of CHB patients, 66.2% of CHC patients, and 62% of CHB+C patients. The character of pathomorphological liver changes in chronic viral hepatitis was studied in 53 renal graft recipients using puncture biopsy. Histopathological activity index (HAI, Knodell R.G. et al., 1981) witnessed a more severe liver lesion in CHB vs. CHC and CHB+C. Thus, inflammatory activity in CHB was found to be minimal or low in 13 patients, and moderate or high in 11 patients, whilst a minimal or low activity in CHC or CHB+C was found in 16 and 10 patients, respectively, and a moderate activity was detected only in two and one, respectively (p = 0.016 and 0.024 compared with CHB). Advanced hepatic fibrosis or cirrhosis was significantly more frequent (p = 0.006) in CHB patients (eight out of 24) than in CHC ones (none out of 18). The rate of advanced sclerotic changes in CHB+C was lower (one out of 10 patients) than that in CHB, and similar to CHC. Thus, clinico-morphological manifestations were more prominent in renal graft recipients with CHB vs. CHC. PMID- 17136856 TI - [Ethicolegal issues of organ donation as the main obstacle to the progress of clinical transplantation]. AB - Ethicolegal issues present the main factor hindering obtaining organs from corpses and living donors, the distribution of cadaveric organs, and the financial providence of donorship in Russia. The actual Russian legislation is contradictory in terms of obtaining relatives' consent for the transplantation of cadaveric organs. There are no precise definitions of the degree of genetic relationship between the donor and recipient sufficient for transplantation. Selling and purchasing human organs is strictly prohibited, while financial compensation of the living donor seems to be a fair measure. Providing the possibility to verify brain death is a necessary condition for expanding the cadaveric organ pool. The organ shortage leads to improper distribution of donor organs. All these problems have to be solved for the progress of organ transplantation in Russia. PMID- 17136857 TI - Chemistry, biosynthesis, analysis, chemical & metabolic transformations and pharmacology. PMID- 17136858 TI - Structure and origin of bile acids: an overview. AB - An overview of the structure and the origin of naturally occurring bile acids is given. Most naturally occurring bile acids belong to the 5beta-series, with hydroxyl groups in the A, B, and C ring of the steroid system. Hydroxyl groups are mostly found at the C3, C6, C7, C12 and C23 positions and are a- rather than beta-oriented. In most bile acids, the A/B ring junction is cis (5beta-series). However, the A ring can be usually present in the more stable (chair) or less stable (boat) conformation. Both B/C and C/D ring junction are trans. With respect to the angular C19-methyl group, the hydrogen atoms at C5 and C8 are cis oriented whereas those at C9 and C14 are trans-oriented. The archetypal bile acid is 5beta-cholanic acid (3) from which all other C24 bile acids can be derived. In addition to the bile acids with 24 carbons, some naturally occurring C27 bile acids have been identified including di-, tri- and tetra-hydroxy derivatives of coprostanic acid isolated from bile of several reptile species. The most dominant bile acids and their natural sources are given and a selection of naturally occurring bile acids with unusual structures which have been mostly isolated from the bile of reptiles and amphibians is described. PMID- 17136859 TI - Biosynthesis of bile acids in mammalian liver. AB - The biosynthesis of bile acids in mammalian liver and its regulation, together with the physiological role of bile acids, are reviewed in this article. Bile acids are biosynthesized from cholesterol in hepatocytes. Several steps are involved including epimerisation of the 3beta-hydroxyl group, reduction of the delta4 double bond to the 5beta-H structural arrangement, introduction of alpha hydroxyl groups at C7 or C7 and C12 and, finally, oxidative degradation of the side chain by three carbon atoms. This gives the primary bile acids, cholic and chenodeoxycholic acids. Cholesterol-7alpha-hydroxylation is the rate determining step in the biosynthesis of cholic and chenodeoxycholic acids. Feedback regulation of cholesterol biosynthesis occurs by various mechanisms including termination of the synthesis of specific cytochromes P-450, modulation of specific cytosol proteins, short-term changes in the process of phosphorylation dephosphorylation and changes in the capacity of the cholesterol pool as a substrate. Prior to being exported from the liver, bile acids are conjugated with glycine and taurine to produce the bile salts. After excretion into the intestinal tract, primary bile acids are partly converted to secondary bile acids, deoxycholic and lithocholic acids, by intestinal microorganisms. The majority of bile acids is absorbed from the intestinal tract and returned to the liver via the portal blood, so that only a small fraction is excreted in the feces. Bile acids returned to the liver can be reconjugated and reexcreted into the bile in the process of enterohepatic recycling. In addition to the physiological function of emulsifying lipids in the intestinal tract, bile acids are particularly important in respect of their ability to dissolve and transport cholesterol in the bile. PMID- 17136860 TI - Isolation and determination of bile acids. AB - In this article, the methods of isolation and determination of bile acids are reviewed. Methods for separation of bile acids from cattle and pig bile are given in detail. Isolation of a mixture of cholic acid and deoxycholic acids from cattle bile and their subsequent purification are described. The isolation and purification of hyodeoxycholic acid and other components of pig bile are also included. Methods for the determination of bile acids in various biological samples are reviewed, including enzyme assays, radioimmunoassay, enzyme immunoassay and chromatographic methods. Among chromatographic methods, separation and determination of bile acids by thin-layer chromatography, gas chromatography and high performance liquid chromatography are reviewed. Particular attention is given to the use of high performance liquid chromatography since this has recently been the most commonly applied method for the separation and determination of bile acids. PMID- 17136861 TI - Chemical and metabolic transformations of selected bile acids. AB - This article surveys chemical transformations of selected bile acids. Chemical transformations were initially carried out with the aim of determining the structure of bile acids. More recently they have been concerned with bile acid interconversions as well as with the synthesis of steroid hormones, vitamins and therapeutc agents. Studies of similarities and differences in the biosynthesis of bile acids from cholesterol have occupied many researches. However, this article reviews only papers dealing with the synthesis of potential intermediates in the biosynthesis of bile acids. Steroid hormones such as pregnenolone, progesterone and testosterone are synthesized from methyl thiodeoxycholate whereas cortisone is synthesized from methyl deoxycholiate. Numerous papers and patents devoted to the synthesis of ursodeoxycholic acid from cholic or chenodeoxycholic acid testify to its effectiveness in the treatment of cholelithiasis. Chenodeoxycholic acid appears to be an excellent precursor in the synthesis of steroid plant growth regulators, as well as in the synthesis of metabolites and vitamin D analogues. Chirality of bile acids has been exploited in the synthesis of cyclic and acyclic receptors and solvents. Cholic and deoxycholic acids have been used to create new macrocyclic structures which show different capacities to bind and transport other compounds. Another important trend in the chemistry of bile acids is their application in combinatorial chemistry. PMID- 17136862 TI - Pharmacology of bile acids and their derivatives: absorption promoters and therapeutic agents. AB - The role of bile acids in pharmacotherapy is reviewed in this article. The therapeutic use of bile has been recognized since ancient times. Previously bile acids were the standard treatment for gallstones where chenodeoxycholic acid and ursodeoxycholic acid were effective in promoting the dissolution of cholesterol gallstones. Today their therapeutic role looks set to expand enormously. Bile acids as absorption promoters have the potential to aid intestinal, buccal, transdermal, ocular, nasal, rectal and pulmonary absorption of various drugs at concentrations that are non-toxic. Keto derivatives of cholic acid, such as 3a,7a,dihydroxy-12-keto-5alpha-cholic acid (sodium salt and methyl ester) are potential modifiers of blood-brain barrier transport and have been shown to promote quinine uptake, enhance the analgesic effect of morphine and prolong the sleeping time induced by pentobarbital. They have also been shown to be hypoglycaemic. Bile acids as therapeutic agents have the potential to produce beneficial effects in sexually transmitted diseases, primary biliary cirrhosis, primary sclerosing cholangitis, gallstones, digestive tract diseases, cystic fibrosis, cancer and diabetes. PMID- 17136863 TI - Costs of outpatient red blood cell transfusions. PMID- 17136864 TI - Pharmacologic therapy for the cancer anorexia/weight loss syndrome: A data driven, practical approach. AB - The cancer anorexia/weight loss syndrome occurs in over 80% of patients with incurable cancer and is associated with a poor prognosis and negative effects on quality of life. Educating patients and families plays an important role in its management, and caregivers sometimes forget that in select patients who are candidates for it, antineoplastic therapy can occasionally reverse some aspects of this syndrome. Patients may also benefit from appetite stimulants such as corticosteroids and progestational agents,and this review summarizes the benefits of using these agents as well as their contraindications. PMID- 17136865 TI - Treating the anorexia/cachexia syndrome. PMID- 17136866 TI - More research needed on the treatment of the cancer anorexia/cachexia syndrome. PMID- 17136867 TI - Community-based clinical oncology research trials for cancer-related fatigue. AB - Cancer-related fatigue (CRF) is a highly prevalent and debilitating symptom experienced by most cancer patients during, and often for considerable periods after, treatment. This symptom affects patients' overall quality of life and is characterized by persistent exhaustion and a decreased capacity to perform daily social and cognitive tasks. CRF is also one of the most commonly reported side effects of cancer and cancer treatments, including surgery, chemotherapy, radiotherapy, hormonal therapy, and biologic response modification therapy. Pharmacologic methods for treating CRF are not always reliable, and many patients continue to experience this symptom following pharmacologic interventions. Managing CRF is of importance to oncology researchers and clinical service providers. As of March 15, 2006, 76 National Institutes of Health sponsored studies examining CRF were actively recruiting subjects, and 15% of these studies were being conducted through the National Cancer Institute (NCI) Community Clinical Oncology Program (CCOP). This article will provide a description of-the NCI CCOP, its functions, and a brief overview of ongoing research on CRF both at our institution and other locations across the United States. PMID- 17136868 TI - Fatigue trials in ambulatory cancer patients:where do we begin and where could it possibly end? PMID- 17136869 TI - Tired of the questions: deciphering cancer-related fatigue. PMID- 17136870 TI - Impact of zoledronic acid on renal function in patients with cancer: Clinical significance and development of a predictive model. AB - Zoledronic acid is a potent bisphosphonate licensed for the treatment of myeloma and bone metastases from solid tumors. Renal deterioration is the most significant toxicity associated with zoledronic acid. We attempted to define the incidence and clinical significance of renal deterioration in patients receiving zoledronic acid and to develop a risk-factor profile for this treatment sequela. This study is a retrospective analysis of all patients who received zoledronic acid at Fox Chase Cancer Center, Philadelphia, Pa, between 1/10/02 and 1/30/04. Data recorded included patient demographics, tumor characteristics, comorbid illnesses, concomitant medications, cancer therapy, number of zoledronic acid doses administered, and serial creatinine measurements. In total, 3,115 evaluable doses of zoledronic acid were administered to 446 patients (median, 4 doses; mean, 6.98 doses; range, 1-28 doses) at a dose of 4 mg over 15 minutes every 3-4 weeks. Of these 446 patients, 42 experienced renal deterioration (median rise in creatinine level, 1.0 mg/dL; range, 0.5-4.4 mg/dL), requiring discontinuation of zoledronic acid therapy in 8 cases. No patient required dialysis and no patient died as a result of zoledronic acid-induced renal dysfunction. On multivariable analysis, predictive factors for the development of renal deterioration were patient age, a diagnosis of myeloma or renal cell cancer, cumulative number of doses, concomitant therapy with a nonsteroidal anti-inflammatory drug, and current or prior therapy with cisplatin. Using these factors, we constructed a predictive model with an area under the receiver operating characteristic curve of 0.75. The incidence of clinically significant renal deterioration in patients treated with zoledronic acid is low.We present a predictive model for decision support when estimating this risk. PMID- 17136871 TI - Cannabinoids in the management of intractable chemotherapy-induced nausea and vomiting and cancer-related pain. PMID- 17136872 TI - Availability and consumption status of CFC and non-CFC inhalers for asthma and chronic obstructive pulmonary diseases in Thailand. AB - In response to the Montreal Protocol and the calls for global early-bird CFC phase-out before 2010, the demand and supply status of both CFC and non-CFC inhalers for prevention and treatment of asthma and COPD in Thailand were evaluated to determine how soon the country would be able to discontinue CFC MDIs with least impacts to both consumers and importers. Availability and supply of the inhalers were collected from registration and importation database of the Thai FDA. Demand and product cost were obtained from the local importers and from IMS, Thailand. Available inhaled products comprise of 39% CFC MDIs, 28% DPIs, 20% solutions for nebulizers and 13% HFA MDls, respectively. All 31 brands of portable hand-held inhalers, comprising 16 CFC MDIs, 6 HFA MDIs and 9 DPIs, are imported, only solutions for nebulization are locally manufactured. Salbutamol is mostly prescribed MDI, its consumption is over 50% of all. The transition to non CFC alternatives (HFA MDIs and DPIs) has become evidence since 2000. After being informed about the demand and supply of the inhalers, in 2005, Thai FDA has announced its CFC phase-out policy and encouraged importation of HFA alternatives by facilitating the registration and approval process. When the most prescribing CFC MDls, salbutamol, is completely replaced with non-CFC form in 2006, Thailand would be able to reduce considerable amount of CFCs into our atmosphere. PMID- 17136873 TI - A randomized controlled trial of cetirizine plus pseudoephedrine versus loratadine plus pseudoephedrine for perennial allergic rhinitis. AB - The purpose of this study was to compare the safety and efficacy of cetirizine plus pseudoephedrine (C+P) with loratadine plus pseudoephedrine (L+P) in the treatment of perennial allergic rhinitis. This was a double blind, randomized, parallel trial with an active control. Subjects aged 12 to 70 years with perennial allergic rhinitis for at least 2 years were enrolled and randomized to receive either of the active study medications plus a placebo resembling the other, twice daily for 4 weeks. Nasal total symptom scale (NTSS) including sneezing, rhinorrhea, nasal itching and nasal stuffiness is evaluated by subjects daily and at baseline, 2 weeks, and 4 weeks by the investigator as efficacy measurement. A total of 51 eligible patients were enrolled and 45 patients completed the treatment course. Both groups had significant reductions in NTSS after 4 weeks of treatment as assessed by the subjects, but there was no significant difference between the two groups (mean +/- SD) reduction of 4.25 +/- 2.45 with C+P vs. 3.52 +/- 2.41 with L+P, p = 0.215. As assessed by the investigator, sneezing was significantly better at 2 weeks (-1.13 vs. -0.52, p = 0.028) and nasal congestion at 4 weeks (-1.71 vs. -1.19, p = 0.031) in subjects treated with C+P compared to those treated with L+P. There were 37 treatment related adverse events (5 in 4 subjects in the C+P group and 32 in 16 subjects in the L+P group). It was concluded that both cetirizine plus pseudoephedrine and loratadine plus pseudoephedrine are efficacious for perennial allergic rhinitis in Taiwanese subjects. Relief of sneezing and nasal congestion may be marginally better with the cetirizine preparation, which also seemed to be slightly better tolerated, although the incidence of side effects did not differ significantly. PMID- 17136874 TI - Incidence of atopic stigmata and prick test results in patients with asthma, allergic rhinitis and conjunctivitis. AB - Allergic diseases are part of the 'modern lifestyle' and their incidence is still increasing. Cutaneous markers (stigmata) usually provide valuable clues for the diagnosis of atopic diseases. This study evaluated the prevalence of the four major and twenty-one minor criteria of Hanifin and Rajka in a total of 246 patients with mucosal allergies (99 asthma, 108 allergic rhinitis, and 39 allergic conjunctivitis). The two most prevalent major criteria were history of atopic diseases and pruritus. The most prevalent three minor criteria were periorbital darkening, influence of environmental factors and xerosis. The most common prick test-positive allergens were grass and mite allergens. Despite evidence for a high co-morbidity between atopic diseases, in daily clinical practise diagnostic and therapeutic procedures generally focus on the most predominant disease. We concluded that it may be important to screen subjects with mucosal allergies for the presence of major and minor cutaneous stigmata. Screening for cutaneous manifestations and subsequent treatment might further enhance the quality of life of these patients. PMID- 17136875 TI - Incidence of anaphylaxis in the emergency department: a 1-year study in a university hospital. AB - The aim of this study was to estimate the incidence of anaphylaxis in an emergency department, identify rate and risk factors of recurrent anaphylaxis, and describe its clinical features and management. A retrospective study of patients who attended the emergency department at Thammasat University Hospital was conducted during 2003-2004 with anaphylactically related ICD-9 and ICD-10 terms. There were 64 patients who experienced 65 anaphylactic episodes during the 1-year period. The anaphylaxis occurrence rate was 223 per 100,000 patients per year. The most common manifestations were cutaneous symptoms and signs, followed by respiratory expression. Food allergy was the most common cause of anaphylaxis. Eighty-five percent of admitted cases had monophasic anaphylaxis. Patients with and without biphasic reactions did not differ significantly in terms of epinephrine and steroid usage. In conclusion, anaphylaxis is not rare. Epinephrine and steroid usage did not prevent biphasic reactions. PMID- 17136876 TI - Evaluation of the nasal provocation test for its necessity in the diagnosis of nasal allergy to house dust mite. AB - The aim of this study was to evaluate rhinomanometric responses to nasal allergen provocation in children with allergic rhinitis sensitized to house dust mite. We studied 51 children, aged 6-16 years (mean: 11.5 +/- 2.6 years), with clinical symptoms of perennial allergic rhinitis without asthma and 20 non-atopic healthy controls in the same age range (mean: 11.8 +/- 3.8 years). All of the patients had positive skin prick test (SPT) results and serum specific IgE above 0.70 kU/l to Dermatophagoides pteronyssinus (Dp). Nasal provocation testing (NPT) was performed with increasing concentrations of Dp extracts and the nasal response was evaluated by active anterior rhinomanometry. A 100% increase of resistance in one or both nasal cavities was considered positive. There was a statistically significant difference of baseline nasal resistance (total, right and left sides) between the control and the patient groups (p < 0.001). A positive response to house dust mite allergens was recorded in 47/51 (92.2%) patients by rhinomanometry. The NPT presented no significant correlation with age, weight, height, SPT diameter, serum total and specific IgE levels to Dp and baseline nasal airway resistance values. This study suggests that a nasal provocation test with allergen is unnecessary in children with positive skin prick test and serum IgE specific to house dust mite. The rhinomanometric response to the allergen provocation does not correlate with the diameter of the skin prick test and the level of serum specific IgE. PMID- 17136877 TI - Computed tomography findings in chronic rhinosinusitis patients with and without allergy. AB - The purpose of this study was to compare the paranasal sinus mucosal thickenings, bony changes consistent with chronic sinusitis, and bony anatomic variations detected by computed tomography (CT) in chronic rhinosinusitis patients with and without allergy. Three hundred and thirty-nine patients with chronic rhinosinusitis were analyzed for their allergic status by performing skin prick test. Two hundred and thirteen patients (62.8%) had at least one positive skin prick test (allergic patients, male/female: 85/128, mean age: 29.1 +/- 1.2). One hundred and twenty-six patients (37.2%) were included in the non-allergic group (male/female: 53/73, mean age: 31 +/- 2.2). Maxillary mucosal thickening and frontal hypoplasy were significantly more common in allergic chronic rhinosinusitis patients. Moreover, pneumatized uncinate process is apparently more common in the allergic group than non-allergic group, and statistical analysis revealed marginal significance (p = 0.0535). In conclusion CT findings of allergic chronic rhinosinusitis patients are comparable to the CT findings of chronic rhinosinusitis patients without allergy. However, presence of maxillary mucosal thickening, frontal hypoplasia or pneumatized uncinate process in the CT scan of a patient with chronic rhinosinusitis could be of clinical significance, and might guide the otolaryngologist for the evaluation of the presence of allergy. PMID- 17136878 TI - Childhood asthma and its relationship with tonsillar tissue. AB - The aim of this study was to evaluate the prevalence of asthma among 4-17 years old school children and to determine the relationship between frequent tonsillitis, tonsillar hypertrophy and asthma-related symptoms. A total of 2,000 school children, aged 4-17 years, in Denizli were surveyed using the ISAAC questionnaire; the size of the tonsils was evaluated by physical examination. A total of 1,784 questionnaires were completed giving an overall response rate of 89.2%. The prevalence of lifetime wheezing, 12-month wheezing and lifetime doctor diagnosed asthma were 16.4%, 6.2% and 1.7%, respectively. The prevalence of frequent tonsillitis and tonsillar hypertrophy were 19.9% and 3.4%, respectively. Tonsillar hypertrophy was positively correlated with a history of frequent tonsillitis. Frequent tonsillitis and tonsillar hypertrophy could be evaluated as risk factors for asthma due to their significant association with asthma-related symptoms. PMID- 17136879 TI - Nocturnal symptoms and sleep disturbances in clinically stable asthmatic children. AB - Presence of nocturnal symptoms is related to asthma severity. Clinically stable asthmatic children, too, report frequent nocturnal symptoms and sleep disturbances. The study determined these parameters in stable, asthmatic children, in their home environment. This case-control, questionnaire-based study in 70 school-going children comprised 40 asthmatics (Group 1) and 30, age/gender matched, healthy children (Group 2). Parents maintained peak expiratory flow (PEF) and sleep diaries for one week. Group 1 had significantly lower mean morning (250.3 vs. 289.1 I/minute) and mean evening PEF values (261.7 vs. 291.3 I/minute). Group 1 (38.95%), reported frequent nocturnal symptoms like cough (36.90%), breathlessness (32.80%), wheeze (27.68%) and chest tightness (14.35%). Sleep disturbances, significant in Group 1 (38, 95% vs. 14.35%), included daytime sleepiness (24.60%), daytime tiredness (20.50%), difficulty in maintaining sleep (15.38%), early morning awakening (14.35%), struggle against sleep during daytime (12.30%), and involuntarily falling asleep (17.43%). On a scale of 1-6, Group 1 scored significant sleep disturbances/patient (3 vs. 0.8); lethargy/tiredness in morning (2.9 vs. 2.2), poorer sleep quality (4.7 vs. 5.4), less parents' satisfaction with child's sleep (4.5 vs. 5.5) and daytime fitness (4.1 vs. 5.3). Group 1, when exposed to environmental tobacco smoke (22, 55%), reported significant nocturnal symptoms (18/22, 81%) and reduced mean morning and evening PEF values (17/22, 77%). It is concluded that clinically stable, asthmatic children reported increased nocturnal symptoms, sleep disturbances and poorer sleep quality. Lack of awareness of asthma-sleep association and its clinical implications could lead to poor asthma control and impaired daytime activity. PMID- 17136880 TI - Three suitable antigens for delayed-type hypersensitivity skin testing in a tropical country like Thailand. AB - Delayed type hypersensitivity (DTH) skin test is a standard tool to assess in vivo cell-mediated immunity. Mantoux method using 4-5 common recalled antigens is recommended. However, not all antigens are widely available and appropriate antigens for tropical countries are not known. The objective of this study is to investigate what and how many antigens should be included in the DTH testing panel that suitable for Thailand and may be for this region. The DTH skin tests were done by Mantoux method in a double blinded fashion. Average induration size of > or = 5 mm defined as a positive test. Antigens included purified protein derivative (PPD), Candida albicans, tetanus toxoid (TT), Trichophyton mentagrophytes and hepatitis B vaccine (HBV). The negative control was normal saline. Of 95 healthy subjects, all showed DTH positive to > or = 1 antigen. The positivity to C. albicans, tetanus toxoid, PPD, T. mentagrophytes, and HBV was 92.6%, 83.2%, 82.1%, 50.5%, and 5.3%, respectively. When three antigens: PPD, TT and C. albicans were analyzed, 100% of subjects showed a positive response to > or = 1 antigen and 96.8% showed a positive response to > or = 2 antigens. When only PPD and TT were analyzed, 100% of subjects showed > or = 1 antigen positive and 68.4% showed both antigens positive. C. albicans antigen at 1:100 was associated with a high incidence of fever (2/20) and large local reaction (7/20), 1:500 was found to be the optimal concentration. PPD, TT and C. albicans are suitable to be included in a DTH skin testing in a tropical country like Thailand. However, in a setting where C. albicans extract is not available, testing with only two antigens of PPD and tetanus toxoid may be an alternative, but with a lower sensitivity. PMID- 17136881 TI - Lack of neutrophil degranulation in low-dose endotoxin inhalation based on a novel intracellular assay. AB - To study the nature of endotoxin or lipopolysaccharide (LPS) induced inflammation, we developed a method of quantifying intracellular human neutrophil elastase (HNE) in lysed sputum polymorphs as a means to study the degranulation status of LPS-recruited neutrophils. Induced sputum, blood and exhaled nitric oxide (NO) were collected from 10 healthy non-atopic human subjects after inhaling a single 15 microg dose of Escherichia coil LPS in an open study. At 6 hours, LPS inhalation caused significant increase of sputum and blood neutrophils but without parallel increase in myeloperoxidase, HNE or interleukin-8 (IL-8) in sputum sol and blood, or exhaled NO. Intracellular HNE in lysed sputum polymorphs or purified blood neutrophils did not show any significant changes between inhaled LPS and saline, nor was there any appreciable change in percentage HNE release induced by N-Formyl-Met-Leu-Phe (fMLP) in vitro. We concluded that in healthy humans, the transient neutrophilic inflammation induced by a single dose of inhaled 15 microg LPS is mainly characterized by cell recruitment, not enhanced secretion of granular mediators or increased exhaled NO based on our experimental conditions. PMID- 17136882 TI - Association of serum levels of tissue inhibitors of metalloproteinase-1 with clinical outcome in children with biliary atresia. AB - The purpose of this study was to determine the possible role of serum levels of tissue inhibitors of metalloproteinase-1 (TIMP-1) in the pathogenesis of the progressive inflammation and fibrosis in biliary atresia (BA). Serum concentrations of TIMP-1 were measured in 57 BA patients and 15 healthy controls using commercially available enzyme-linked immunosorbent assays. The mean ages of the BA patients and the controls were 6.1 +/- 0.6 and 6.7 +/- 1.1 years, respectively. The patients were categorized into two groups according to their clinical outcomes: patients with jaundice (total bilirubin > or = 2 mg/dl) and patients without jaundice (total bilirubin < 2 mg/dl). In our study, serum levels of TIMP-1 were significantly higher in the BA patients than in healthy subjects (4.8 +/- 0.4 vs. 3.5 +/- 0.3 ng/ml, respectively; p < 0.05). Additionally, serum levels of TIMP-1 significantly increased in the BA patients with jaundice in comparison to those without jaundice (6.3 +/- 0.7 vs. 3.1 +/- 0.3 ng/ml, respectively; p = 0.001). Patients with persistent jaundice had lower levels of albumin but had greater levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and gamma glutamyl transpeptidase compared with patients without jaundice. Furthermore, patients with portal hypertension (PH) had higher TIMP-1 levels than those without PH (5.3 +/- 0.4 vs. 1.9 +/- 0.3 ng/ml, respectively; p < 0.001). It is concluded that serum levels of TIMP-1 increased in patients with BA. The significant increase in TIMP-1 levels is related to the presence of PH and the severity of jaundice. The elevated TIMP 1 levels may reflect the degree of hepatic fibrosis and development of PH. The data suggest that TIMP-1 may play a role in the pathophysiology of post-Kasai BA. PMID- 17136883 TI - Buckwheat anaphylaxis: an unusual allergen in Taiwan. AB - IgE-mediated hypersensitivity to buckwheat is common in Korea, Japan, and some other Asian countries. However, buckwheat is not a common allergen in Taiwan. We report a woman with asthma who had anaphylactic shock, generalized urticaria, and an acute exacerbation of asthma five minutes after ingesting buckwheat. The patient underwent skin prick and Pharmacia CAP testing (Uppsala, Sweden) for specific IgE to buckwheat, white sesame and soybean as well as other common allergens in Taiwan including Dermatophagoides pteronyssinus (Dp), D. farinae (Df), cat and dog dander, cockroach, egg white, cow milk and codfish. The patient had a strongly positive skin prick test response to buckwheat and positive reactions to Dp and latex. Specific IgE results were class 6 for buckwheat, class 4 for Dp and Df, and class 2 for dog dander, wheat, sesame and soybean. Results of an open food challenge with white sesame and soybean were negative. Although buckwheat is a rare allergen in Taiwan, it can cause extremely serious reactions and should be considered in patients presenting with anaphylaxis after exposure to buckwheat. PMID- 17136884 TI - Recurrent Campylobacter lari bacteremia in X-linked agammaglobulinemia: a case report and review. AB - X-linked agammaglobulinemia (XLA) is a primary immune deficiency disease with a B cell defect. We present the first XLA patient who had recurrent Campylobacter lari bacteremia. High dose intravenous immunoglobulin combined with azithromycin once per week, and a complete avoidance of bacterial reservoirs may be helpful for the prevention of C. lari bacteremia. PMID- 17136885 TI - Aminoglycosides: ancient and modern. PMID- 17136886 TI - Genomic analyses lead to novel secondary metabolites. Part 3. ECO-0501, a novel antibacterial of a new class. AB - Genomic analyses of Amycolatopsis orientalis ATCC 43491 strain, deposited as a vancomycin producer, revealed the presence of genetic loci for the production of at least 10 secondary metabolites other than vancomycin. One of these gene clusters, which contained a type I polyketide synthase, was predicted to direct the synthesis of novel class of compound, a glycosidic polyketide ECO-0501 (1). Screening of culture extracts for a compound with the predicted physicochemical properties of the product from this locus, led to the isolation of the 13-O glucuronide of 13-hydroxy-2,12,14,16,22-pentamethyl-28-(N-methyl-guanidino) octacosa-2,4,6,8,10,14,20,24-octaenoic acid (2-hydroxy-5-oxo-cyclopent-1-enyl) amide (ECO-0501, 1). The structure, confirmed by spectral analyses including MS, and ID and 2D NMR experiments, were in accord with that predicted by genomic analyses. ECO-0501 possessed strong antibacterial activity against a series of Gram-positive pathogens including several strains of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE). ECO-0501 was chemically modified by esterification (1a-1c), N-acetylation (1d) and hydrogenation (1e) in order to explore structure activity relationships (SAR). PMID- 17136887 TI - Importance of the structure of vancomycin binding pocket in designing compounds active against vancomycin-resistant enterococci (VRE). AB - 16-Membered meta, para-cyclophanes mimicking the vancomycin binding pocket (D-O-E ring) are designed and synthesized. The structural features of these biaryl ether containing macrocycles are: a) the deletion of the carboxyl group of vancomycin's central amino acid (amino acid D); b) the elongation of the N-terminal; c) the presence of lipidated aminoglucose at the D-ring. Cycloetherification by way of an intramolecular nucleophilic aromatic substitution reaction (S(N)Ar) is used as a key step for the construction of the macrocycle. Minimum inhibitory concentrations for all of the derivatives are measured using a standard microdilution assay. Compounds 2a-2c and 3a-3c displayed weak activities against resistant strain Enterococcus faecalis L560 and were inactive against Enterococcus faecium resistant strain L2215. PMID- 17136888 TI - New destruxins from the marine-derived fungus Beauveria felina. AB - Chemical investigation of the cytotoxic and anti-tuberculosis active butanone extract obtained from the growth media of the marine-derived fungus Beauveria felina led to the isolation of two new destruxins, [beta-Me-Pro] destruxin E chlorohydrin (1) and pseudodestruxin C (3), along with five known cyclic depsipeptides. The structures of the new destruxin derivatives were established by analysis of spectroscopic data, while the absolute configuration of the common amino acid residues was established by Marfey's analysis. The absolute configuration of the 2(R),4(S)-5-chloro-2,4-dihydroxypentanoic acid residue in 1 could be established by application of a J-based configuration method followed by derivatization with R-MPA-Cl and NMR analysis. PMID- 17136889 TI - A new series of glycopeptide antibiotics incorporating a squaric acid moiety. Synthesis, structural and antibacterial studies. AB - The aglycones of the antibiotics eremomycin, vancomycin and ristocetin (3, 4 and 6, respectively) were prepared by deglycosidation of the parent antibiotics with hydrogen fluoride, and complete assignation of their 1H, 13C and 15N spectra was performed. The squaric acid amide esters (11-14), were prepared from dimethyl squarate. The corresponding asymmetric diamides (16-19, 22, 23) were also synthesized using 4-phenylbenzylamine and triglycine. The advantage of the method is the high regioselectivity and that no protecting group strategy is required. Electrospray mass spectroscopic method was elaborated for the determination of the site of substitution of the modified antibiotics. The antibacterial activity of the prepared compounds is discussed in detail. PMID- 17136890 TI - Antimalarial activity of crambescidin 800 and synthetic analogues against liver and blood stage of Plasmodium sp. AB - Structural features associated with the antimalarial activity of the marine natural product crambescidin 800 were studied using synthetic analogues of the related compound ptilomycalin A. The study suggests that the guanidine moiety is cytotoxic, whereas the spermidine-containing aliphatic chain increases activity. The most active analogue, compound 11, had in vitro activity against Plasmodium falciparum strain 3D7 (IC50=490 nM) that was stronger than the in vitro activity against murine L5178Y cells (IC50 = 8.5-59 microM). In vitro growth inhibition of liver stages of P. yoelii yoelii in mouse hepatocytes was observed (IC50 = 9.2 microM). The compound did not significantly prolong median survival time after a single subcutaneous administration of 80 mg/kg in P. berghei-infected mice. Compound 11 did not cause DNA fragmentation in an in vitro micronucleus assay. PMID- 17136891 TI - Paecilaminol, a new NADH-fumarate reductase inhibitor, produced by Paecilomyces sp. FKI-0550. AB - A new NADH-fumarate reductase inhibitor, paecilaminol, was isolated from the cultured broth of a fungus Paecilomyces sp. FKI-0550. It is an amino alcohol compound, the structure being established as 2-amino-14,16-dimethyl-3 octadecanol. Paecilaminol exhibited an IC50 value of 5.1 microM against Ascaris suum NADH-fumarate reductase. PMID- 17136892 TI - A new antifungal macrolide, eushearilide, isolated from Eupenicillium shearii. AB - In screening for antifungal substances, a new macrolide, eushearilide (1), was isolated from Eupenicillium shearii IFM54447. The structure of 1 was established to be 24-membered macrolide having a non-conjugated diene and a choline phosphate ester moetiy on the basis of detailed investigation of NMR, UV, IR and MS spectral data. Compound 1 showed antifungal activity against various fungi and yeasts, including human pathogens Aspergillus fumigatus, Trichophyton spp. and Candida spp. PMID- 17136893 TI - Studies on novel bacterial translocase I inhibitors, A-500359s. V. Enhanced production of capuramycin and A-500359 A in Streptomyces griseus SANK 60196. AB - Streptomyces griseus SANK 60196 produces the novel nucleoside antibiotics A 500359 A, C, D and capuramycin. Enhanced production of capuramycin and A-500359 A was achieved through a number of medium modifications and a series of single colony isolations. The addition of maltose instead of glucose as the carbon source in a primary medium resulted in a 20-fold increase in the productivity of capuramycin. Furthermore, the addition of cobalt chloride (CoCl2) and yeast extract to the medium containing maltose drastically altered the production ratio of A-500359 A to capuramycin. Thus, the yield of A-500359 A increased up to 600 microg/ml in an optimal medium, while the yield in the primary medium was 1 microg/ml. PMID- 17136894 TI - Biosynthetic study of FR-900848: origin of the aminodeoxynucleoside part. AB - Biosynthetic studies of the antifungal agent, FR-900848, were undertaken by feeding experiments with D-[U-13C,]glucose, L-[4-13C]aspartate, [5,5 2H,]dihydrouridine and [5,5-2H2]dihydrouracil. The 5"-amino-5"-deoxy-5',6' dihydrouridine moiety was derived from ribose and aspartate. Based on the feeding experiments, a detailed biosynthetic pathway producing the aminodeoxydihydrouridine moiety of FR-900848 was proposed. PMID- 17136895 TI - The more things change... PMID- 17136896 TI - The routine offer of HIV counseling and testing: a human right. PMID- 17136897 TI - Routine HIV testing: a view from Botswana. PMID- 17136898 TI - The International Community of Women Living with HIV/AIDS: point of view. PMID- 17136899 TI - Scaling up HIV testing: ethical issues. PMID- 17136900 TI - HIV testing in the era of treatment scale up. PMID- 17136901 TI - HIV testing: breaking the deadly cycle. PMID- 17136902 TI - Ensuring a rights-based approach to HIV testing. PMID- 17136903 TI - Prisoners who inject drugs: public health and human rights imperatives. AB - This article examines the human rights and public health implications of injection drug use in prisons with a specific focus on HIV and hepatitis C (HCV) viruses. The authors argue that prisoners who inject drugs have a right to access harm reduction measures--those that reduce the harmful consequences of drug use without necessarily reducing drug consumption. Moreover, states that fulfill their obligation to provide prisoners with harm reduction measures such as access to bleach, substitution therapy, and sterile injection equipment implement sound public health policy with a positive impact for a population particularly vulnerable to HIV and HCV. Ultimately, this approach benefits not only prisoners but also prison staff and the public, and does not entail lessening of the safety and security of prisons. PMID- 17136904 TI - On a never-ending waiting list: toward equitable access to anti-retroviral treatment? Experiences from Zambia. AB - Universal access to anti-retroviral (ARV) medication for HIV/AIDS is the clarion call of the WHO/UNAIDS 3 by 5 Initiative. Treatment coverage, however, remains highly uneven. This sharpens the question of who exactly is accessing ARVs and whether access is challenging inequality or reinforcing it. Issues of distributive justice have long been debated in health policy, but the practical challenges of ARV distribution are relatively new. In exploring what a more equitable process of ARV distribution could involve, this article draws on a human rights framework using case study material from Zambia. PMID- 17136905 TI - Harm reduction, HIV/AIDS, and the human rights challenge to global drug control policy. AB - The global HIV/AIDS pandemic, and the role of unsafe drug injection as one of its principal drivers, have added to the list of harms associated with unsafe drug use. HIV/AIDS has highlighted ways in which prohibitionist drug policy causes or contributes to such harms and focused attention on the international regime of illicit drug control. At the same time, HIV/AIDS has catalyzed the "health and human rights movement" to articulate legal and policy responses that both represent sound public health policy and fulfill human rights obligations recognized in international law; this necessarily includes scrutinizing the interpretation and implementation of the UN drug control conventions. This article brings together public health evidence and legal analysis as a contribution toward changing the global drug control regime to a more health friendly, human rights-based system. PMID- 17136906 TI - HIV and domestic violence: intersections in the lives of married women in India. AB - Gender inequality is driving two distinct yet interlinked epidemics among women in India: HIV and AIDS and domestic violence. As domestic violence is increasingly recognized and HIV infection expands, policy and programs do not reflect the interlinked risks and consequences in married women's lives. This article seeks to establish the nexus between HIV and AIDS and domestic violence and identify potential areas for a state-led response. In a health and human rights approach, it assesses women's vulnerability to each epidemic at the individual, societal, and program levels to analyze direct and underlying factors that determine women's risk. Three areas are identified as opportunities for an integrated response: strengthen HIV and domestic violence strategies and address their overlap; mainstream gender; and improve data and research. PMID- 17136907 TI - Getting global funds to those most in need: the Thai Drug Users' Network. AB - The Thai Drug Users' Network (TDN) formed in 2002 in response to the deplorable health and human rights conditions facing illicit drug users in Thailand. In 2003, TDN submitted a proposal for funding to the Global Fund to Fight AIDS, Tuberculosis, and Malaria (GFATM). The proposal appeared destined to fail as it did not have the support of the Thai government and because the interventions proposed were highly incongruent with national policies. In the midst of an ongoing drug war, TDN took action to increase awareness of the harms of the drug war, the shortcomings in GFATM policies, and the need for peer-driven interventions. This article describes the circumstances surrounding this instance of successful drug-user organizing in order to identify the lessons that can be learned. PMID- 17136908 TI - East-West fusion: cross-border human rights activism and the Thai Drug Users' Network. AB - Recently, international human rights leaders have renewed the call for advocacy on economic, social, and cultural rights and suggested partnerships with local organizations. The Thai Drug Users' Network (TDN) promotes the human rights of a marginalized and medically underserved population within Thailand. It also works internationally to reduce drugrelated harms. Thus, TDN transcends a strict local international dichotomy. The group grew out of professional and personal ties between Thai drug users and international health and human rights actors. Border crossing connections and two-way transfer of knowledge, particularly through bridging individuals or "cultural translators," have benefited both TDN and the non-Thai organizations that work with it. This case study shows how international local, and even donor-recipient, relationships may be navigated in ways that are symbiotic and mutually empowering. PMID- 17136909 TI - Rights matter: structural interventions and vulnerable communities. PMID- 17136910 TI - HIV/AIDS and individuals with disability. PMID- 17136911 TI - International Community of Women Living with HIV/AIDS. PMID- 17136912 TI - Service framework for long-term conditions. PMID- 17136913 TI - Preparing a healthcare workforce for the 21st century: the challenge of chronic conditions. PMID- 17136914 TI - Is depression a chronic illness? For the motion. PMID- 17136915 TI - Is depression a chronic illness? Parker opposes the motion. PMID- 17136916 TI - Is depression a chronic illness? A response from the perspective of general practice. PMID- 17136917 TI - Is depression a chronic illness? A sociological perspective. PMID- 17136918 TI - The impact of social deprivation on chronic back pain outcomes. AB - Chronic low back pain is the commonest cause of disability for adults of working age. It is a complex problem frequently encapsulated as a bio-psychosocial issue, yet the social element has received less attention than it deserves, particularly for low-income and socially deprived patients. Rehabilitation programmes are often based on increasing function through cognitive and behavioural techniques, which, for many reasons, may be less effective for the socially disadvantaged. In this paper we discuss the potential barriers to successful rehabilitation in socially deprived groups and we look at possible factors that may need to be considered when designing interventions. PMID- 17136919 TI - Measuring outcomes for neurological disorders: a review of disease-specific health status instruments for three degenerative neurological conditions. AB - Health-related quality-of-life measures have been increasingly used in research into neurological disorders in recent years. The aim of this paper is to provide an objective appraisal of the evidence in regard to disease-specific quality-of life measures used in research on health interventions for three degenerative neurological disorders: multiple sclerosis, motor neurone disease/amyotrophic lateral sclerosis and Parkinson's disease. A comprehensive search strategy was developed to include nine relevant electronic databases. Only studies pertaining to patient-based outcome measurements in multiple sclerosis, motor neurone disease and Parkinson's disease were included. We identified 76 eligible studies. As studies consisted of descriptive and cross-sectional survey study designs, results were reported qualitatively rather than in the form of a meta-analysis. Four disease-specific measures were found for Parkinson's disease, 11 for multiple sclerosis and one for motor neurone disease. We conclude that health related quality-of-life measures are useful in assessing the impact of treatments and interventions for neurological disorders. However, further research is needed on the development of instruments using psychometric methods and on the validation, utilization and responsiveness of instruments to change. PMID- 17136920 TI - Harnessing the potential of the internet to promote chronic illness self management: diabetes as an example of how well we are doing. AB - OBJECTIVES: Given the potential for the Internet to be used as a dynamic, interactive medium for providing information, changing attitudes and behaviour and enhancing social support, it is important to consider whether what is currently available online for chronic illness self-management adequately harnesses this potential. The objective of this paper was to review the content of diabetes self-management websites and to identify strengths and limitations of online diabetes self-management. METHODS: We reviewed and coded features of 87 publicly available diabetes websites hosted by governmental, health plan, commercial, pharmaceutical, and not-for-profit organizations. We assessed whether each website was using online opportunities in the areas of interactivity, theory based interventions, social support, and evidence-based care. RESULTS: The majority of sites provided information, essentially using an electronic newspaper or pamphlet format. Few sites offered interactive assessments, social support or problem-solving assistance, although there were some significant differences in these characteristics across the types of site. DISCUSSION: Current diabetes websites fall short of their potential to help consumers. Suggestions are made for ways to improve the helpfulness and interactivity of these resources. PMID- 17136921 TI - Pastoral relationships and holding work in primary care: affect, subjectivity and chronicity. AB - OBJECTIVES: To understand family doctors' constructs of long-term therapeutic relationships with patients in primary care. METHODS: Semi-structured interviews were administered to general practitioners with > 5 years of experience (n = 28) working in an English semi-rural district, and the results were subjected to constant comparative qualitative analysis. RESULTS: Participants identified pastoral relationships as long-standing patterns of doctor-patient interaction aimed at providing reliable supportive care indirectly concerned with clinical medicine. Holding work was identified as a technique for structuring and delivering care within pastoral relationships. Pastoral relationships and holding work were seen as valuable in the affective management of people with long standing chronic illness, especially mild-to-moderate depression and anxiety. DISCUSSION: At a time when primary care is undergoing significant structural change, respondents in this study laid emphasis on personal and continuing relationships with patients who had diffuse needs connected with the experience of complex and chronic problems, and their accounts intimately connected life events with health status. Importantly, these accounts suggest that such relationships are hard to define and therefore hard to measure, but have important therapeutic purposes. PMID- 17136922 TI - Evidence of cultural hybridity in responses to epilepsy among Pakistani muslims living in the UK. AB - OBJECTIVES: To examine how people from Bradford's Pakistani Muslim community experience living with epilepsy. Specifically, the paper addresses social interactions and negotiations with care providers and considers how different understandings of epilepsy are integrated. METHODS: Interviews were conducted with a sample of Bradford's Pakistani Muslim community (n = 20). Interviews were analysed to identify themes and significant areas of shared concern. RESULTS: This paper identifies popular, professional and folk sectors contributing to an individual's 'health system'. Where sectors overlap, zones of hybridity are created: that is, a person might simultaneously seek help from a doctor and from a religious healer, or might offer explanations for seizures that include neurological and spiritual components. DISCUSSION: While there are many similarities between the experiences of these minority ethnic community members and published work on the lived experience of epilepsy in other communities, there are also important differences that service providers need to recognize and respond to. Differences include forms of cultural expression and specific language needs. Improving communication between professionals and persons with epilepsy needs to be prioritized. PMID- 17136923 TI - What is chronic illness? PMID- 17136924 TI - Maze migraine survey. PMID- 17136925 TI - Epilepsy pregnancy registers. PMID- 17136926 TI - On quality. PMID- 17136927 TI - Chronic illness and intractability: professional-patient interactions in primary care. PMID- 17136928 TI - A response to intractability--a way forward. PMID- 17136929 TI - The personal and political dimensions of 'chronic disease'. PMID- 17136930 TI - Chronic illness: epidemiological or social explosion? PMID- 17136931 TI - A trial of asthma self-management in Beijing schools. AB - OBJECTIVES: This study examined the effectiveness in children in China of an asthma education programme adapted from a model developed in the USA. METHODS: Six hundred and thirty-nine children in 21 elementary schools in one agricultural and one industrial area participated in a randomized, controlled trial. Data were collected at baseline and 1 year subsequently. The self-regulation-based programme addressed topics including preventing and managing symptoms, using medicines, and identifying and controlling triggers. RESULTS: Positive effects on treatment children v. control children were noted in school performance (0.21 v. 0.06, p=0.04), absences (-0.55 v. -0.32, p= 0.02), and home environment (1.78 v. 4.75, p= 0.009). Industrial-area children additionally benefited from fewer hospitalizations (odds ratio =1.96, p =0.05) and asthma-related concerns of parents (-0.63 v. -0.34, p = 0.001). Agricultural-area parents showed greater improvement in asthma management (0.93 v. 0.26, p= 0.0001), and expressed more negative feelings about asthma (-0.13 v. - 0.58, p= 0.04) and asthma concerns ( 0.31 v. -0.63, p= 0.0001). DISCUSSION: The programme provided overall benefits related to school performance, absences, and home environment. In the agricultural area, where fewer resources were available, benefits were fewer and concerns greater. In the industrial area, where education and income were higher, additional benefits related to healthcare use and parents' quality of life were realized. PMID- 17136932 TI - Patients' and nurses' views of nurse-led heart failure clinics in general practice: a qualitative study. AB - OBJECTIVES: To ascertain nurses' and patients' views and experiences of a nurse led heart failure clinic provided in general practice. METHODS: The study was set in eight general practices in the North-West of England. Semi-structured interviews were devised and administered, with all the nurses providing the clinics and a purposive sample of patients attending the clinics. The interviews were tape-recorded and transcribed. Constant comparative analysis was used to identify key issues and themes. RESULTS: Nurses felt that the self-care advice provided had empowered patients to manage their condition. Explaining why a medication had been prescribed, and how it controlled heart failure, was felt to increase compliance. Although communication was deemed good, some patients were reticent about asking questions. Patients were knowledgeable about their prescribed heart failure medications, but some did not recall having discussed their medications. Also, medication inserts led some patients to question their prescription. Patients remained confused about the purpose and outcome of investigations. Furthermore, many patients suggested that they had problems adhering to or remembering the advice given. DISCUSSION: There are practical benefits to be obtained from attending a nurse-led heart failure clinic in primary care. However, patients and healthcare providers may have quite divergent views about such a service and its benefits, emphasizing the potential value of consumer involvement and feedback when developing and delivering such services. PMID- 17136933 TI - Chronically ill rural women: self-identified management problems and solutions. AB - OBJECTIVE: To add to the knowledge base of illness management of chronically ill, rural women by describing the self-identified problems and solutions reported by women participants in the online health-education segment of the Women to Women (WTW) computer outreach project. METHODS: WTW is a research-based computer intervention providing health education and online peer support for rural women with chronic diseases. Messages posted to the online chat room were examined to determine the women's self-management problems and solutions. RESULTS: The self identified problems were: (1) difficulties in carrying through on self-management programmes; (2) negative fears and feelings; (3) poor communication with care providers; and (4) disturbed relationships with family and friends. The self identified solutions to these problems included problem-solving techniques that were tailored to the rural lifestyle. Although not all problems were 'solvable', they could be 'lived with' if the women's prescriptions for self-management were used. DISCUSSION: Glimpses into the women's day-to-day experiences of living with chronic illness gleaned from the interactive health-education discussions will give health professionals insights into the women's efforts to manage their illnesses. The data provide health professionals with information to heighten their sensitivity to their clients' day-to-day care and educational needs. PMID- 17136934 TI - Women's and doctors' accounts of their experiences of depression in primary care: the influence of social and moral reasoning on patients' and doctors' decisions. AB - BACKGROUND: Little is known about how general practitioners (GPs) manage depression within everyday clinical practice, particularly in relation to the issue of 'problem definition'. In addition, there has been relatively little research on the patients' perspective of depression and its management in primary care. METHODS: Qualitative interviews explored women's and GPs' experiences of recognizing depression and their experiences of the management of depression. Thirty-seven women and 20 GPs were recruited from practices in four National Health Service Board areas of Scotland. Each participant was interviewed at the start of the study, and 30 women and 19 GPs were revisited approximately 9-12 months later so that the process of care could be reviewed. RESULTS: The findings demonstrate the social and moral reasoning that lies behind women's decisions to seek help and to subsequently accept their GPs' explanation and advice, and that the acceptance of antidepressants created a moral dilemma for the women. For GPs, the diagnosis and management of depression led to contemplating the boundaries of their professional role, and social and moral reasoning was also evident in their decision-making processes. DISCUSSION: The implication is that, for the majority of women, a chronic-disease model for the management for depression in primary care would be likely to increase rather than reduce the moral dilemma. In addition, the management of depression is not solely based on clinical decisions, so the applicability of a chronic-disease model to primary care requires further consideration. PMID- 17136935 TI - Chronic disease management in Australia: evidence and policy mismatch, with asthma as an example. AB - All Western countries are developing and implementing new models for managing asthma. In Australia, the Asthma 3+ Visit Plan was implemented in 2001-2002, with the aim of establishing a structured approach in general practice for people with moderate and severe asthma, within a fee-for-service system. Evidence is emerging that the uptake across general practice has been poor, with approximately 5% of eligible people being enrolled. The reasons for the poor uptake include such factors as an inability to identify people with asthma, general practitioners being too busy with other clinical priorities, and the lack of interest and understanding of the need for a structured approach for this disease. This mismatch between evidence and policy development needs to be rectified, if sustainable models of chronic disease care are to be firmly established in general practice. PMID- 17136936 TI - Can interprofessional education make a difference in the care of people with chronic disease? AB - In recent years, there has been an increasing policy focus on the role of interprofessional education in healthcare. However, the literature emphasizes process rather than outcomes, with little that specifically relates interprofessional educational interventions to the care of people with long-term conditions. This article questions what we know about interprofessional interventions in the care of people with chronic disease, examines the evidence for and potential of interprofessional education in changing practice, and makes suggestions for further research. PMID- 17136937 TI - Passive smoking and chronic illness in children: age and gender inequalities, and the fallacy of 'low-strength' cigarettes. PMID- 17136938 TI - Exercise, nutrition, and homocysteine. AB - Homocysteine is an independent cardiovascular disease (CVD) risk factor modifiable by nutrition and possibly exercise. While individuals participating in regular physical activity can modify CVD risk factors, such as total blood cholesterol levels, the impact physical activity has on blood homocysteine concentrations is unclear. This review examines the influence of nutrition and exercise on blood homocysteine levels, the mechanisms of how physical activity may alter homocysteine levels, the role of homocysteine in CVD, evidence to support homocysteine as an independent risk factor for CVD, mechanisms of how homocysteine increases CVD risk, and cut-off values for homocysteinemia. Research examining the impact of physical activity on blood homocysteine levels is equivocal, which is partially due to a lack of control for confounding variables that impact homocysteine. Duration, intensity, and mode of exercise appear to impact blood homocysteine levels differently, and may be dependent on individual fitness levels. PMID- 17136939 TI - Protein intake for skeletal muscle hypertrophy with resistance training in seniors. AB - Variability in protein consumption may influence muscle mass changes induced by resistance exercise training (RET). We sought to administer a post-exercise protein supplement and determine if daily protein intake variability affected variability in muscle mass gains. Men (N=22) and women (N=30) ranging in age from 60 to 69 y participated in a 12-wk RET program. At each RET session, participants consumed a post-exercise drink (0.4 g/kg lean mass protein). RET resulted in significant increases in lean mass (1.1 +/- 1.5 kg), similar between sexes (P > 0.05). Variability in mean daily protein intake was not associated with change in lean mass (r < 0.10, P > 0.05). The group with the highest protein intake (1.35 g x kg(-1) x d(-1), n=8) had similar (P > 0.05) changes in lean mass as the group with the lowest daily protein intake (0.72 g x kg(-1) x d(-1), n=9). These data suggest that variability in total daily protein intake does not affect variability in lean mass gains with RET in the context of post-exercise protein supplementation. PMID- 17136940 TI - Increased dietary protein and combined high intensity aerobic and resistance exercise improves body fat distribution and cardiovascular risk factors. AB - We investigated the effectiveness of two lifestyle modification programs of exercise training and nutritional intake (ad libitum) on improving body composition and disease risk in overweight/obese men and women. Sixty-three subjects were weight matched and assigned to one of three groups for a 12 wk intervention: (1) high-intensity resistance and cardiovascular training and a balanced diet (RC+BD, 40% CHO: 40% PRO; n=27, 16 female/11 male, age = 42 +/- 9 y); (2) moderate-intensity cardiovascular training and a traditional food guide pyramid diet (C+TD, CHO 50 to 55%; PRO 15 to 20%; FAT < 30%; n=19, 10 female/9 male, age = 43 +/- 10 y); and (3) an inactive control group (C, n=17, 5 female/12 male, age 43 +/- 11 y). RC+BD resulted in more favorable changes (P < 0.01) in percent body fat (-15.8% vs. -6.9%) and abdominal fat (-15.6% vs. -7.5%) compared to C+TD and C. Total cholesterol (-13.8%), LDL-cholesterol (-20.8%), and systolic blood pressure (-5.7%) declined (P > 0.05) in RC+BD, whereas C+TD and C remained unchanged. Our results suggest that RC+BD may be more effective than C+TD and C in enhancing body composition and lowering cardiovascular risk in obese individuals. PMID- 17136941 TI - The effect of high carbohydrate meals with different glycemic indices on recovery of performance during prolonged intermittent high-intensity shuttle running. AB - This study examined the effect of high carbohydrate meals with different glycemic indices (GI) on recovery of performance during prolonged intermittent high intensity shuttle running. Seven male semi-professional soccer players (age 23 +/ 2 y, body mass [BM] 73.7 +/- 9.0 kg and maximal oxygen uptake 58 +/- 1.0 mL x kg(-1) min(-1)) participated in two trials in a randomized cross-over design. On day 1, the subjects performed 90 min of an intermittent high-intensity shuttle running protocol [Loughborough Intermittent Shuttle Test (LIST)]. They then consumed a mixed high carbohydrate recovery diet (8 g/kg BM) consisting of either high (HGI) (GI: 70) or low (LGI) (GI: 35) GI foods. Twenty-two hours later (day 2) the subjects completed 75 min of the LIST (part A) followed by alternate sprinting and jogging to fatigue (part B). No differences were found between trials in time to fatigue (HGI 25.3 +/- 4.0 min vs. LGI 22.9 +/- 5.6 min, P = 0.649). Similarly, no differences were found between trials for sprint performance and distance covered during part B of the LIST. In conclusion, the GI of the diet during the 22 h recovery did not affect sprint and endurance performance the following day. PMID- 17136942 TI - Dietary antioxidant supplementation combined with quercetin improves cycling time trial performance. AB - We investigated whether 6 wk of antioxidant supplementation (AS) would enhance 30 km time trial (TT) cycling performance. Eleven elite male cyclists completed a randomized, double-blind, cross-over study to test the effects of twice daily AS containing essential vitamins plus quercetin (FRS), and AS minus quercetin (FRS Q) versus a baseline TT (B). MANOVA analysis showed that time to complete the 30 km TT was improved by 3.1% on FRS compared to B (P < 0.01), and by 2% over the last 5 km (P < 0.05). Absolute and relative (%HRmax) heart rates and percent VO2max were not different between trials, but average and relative power (% peak power) was higher on FRS (P < or = 0.01). Rates of carbohydrate and fat oxidation were not different between trials. Thus, FRS supplementation significantly improved high-intensity cycling TT performance through enhancement of power output. Further study is needed to determine the potential mechanism(s) of the antioxidant efficacy. PMID- 17136943 TI - Protein added to a sports drink improves fluid retention. AB - The purpose of this study was to compare fluid retention of carbohydrate plus protein, a carbohydrate-only, and water following 2.5% body weight (BW) loss. Thirteen subjects dehydrated to 2.5% of BW, then ingested a CHO (6%) plus protein drink (1.5%; CP), a 6% CHO drink, or water (WA) at a volume equal to BW loss during a 3-h recovery. Fluid retention was significantly greater for CP (88 +/- 4.7%) than CHO (75 +/- 14.6%), which was greater than WA (53 +/- 16.1%). Serum and urine osmolalities were greater for CP (284.7 +/- 5.0; 569.4 +/- 291.4 mOsm/kg) than CHO (282.6 +/- 5.2; 472.9 +/- 291.5 mOsm/kg) which were greater than WA (280.6 +/- 5.9, 303.7 +/- 251.5 mOsm/kg). Results indicate that fluid retention for CP was 15% greater than CHO and 40% greater than WA. Water ingestion led to a dilution of the serum and resulted in only 53% fluid retention. PMID- 17136944 TI - Effect of creatine and beta-alanine supplementation on performance and endocrine responses in strength/power athletes. AB - The effects of creatine and creatine plus beta-alanine on strength, power, body composition, and endocrine changes were examined during a 10-wk resistance training program in collegiate football players. Thirty-three male subjects were randomly assigned to either a placebo (P), creatine (C), or creatine plus beta alanine (CA) group. During each testing session subjects were assessed for strength (maximum bench press and squat), power (Wingate anaerobic power test, 20 jump test), and body composition. Resting blood samples were analyzed for total testosterone, cortisol, growth hormone, IGF-1, and sex hormone binding globulin. Changes in lean body mass and percent body fat were greater (P < 0.05) in CA compared to C or P. Significantly greater strength improvements were seen in CA and C compared to P. Resting testosterone concentrations were elevated in C, however, no other significant endocrine changes were noted. Results of this study demonstrate the efficacy of creatine and creatine plus beta-alanine on strength performance. Creatine plus beta-alanine supplementation appeared to have the greatest effect on lean tissue accruement and body fat composition. PMID- 17136945 TI - Anxiety and depression in COPD: a call (and need) for further research. PMID- 17136946 TI - Effect of bilateral lung volume reduction surgery on FEV1 decline in severe emphysema. AB - STUDY OBJECTIVE: To examine whether lung volume reduction surgery (LVRS) alters the anticipated natural rates of decline in FEV1. DESIGN: Retrospective analysis of spirometry results (188 studies) in patients before and after bilateral LVRS. Setting. Large, urban, academic medical center. PATIENTS: 25 patients with severe emphysema (mean (SD) age 60+/-8 yrs; FEV1 0.74+/-0.29 L, 29% predicted). INTERVENTIONS: Bilateral LVRS performed via median sternotomy, with areas targeted for resection based on preoperative evaluation using high-resolution computed tomography, quantitative perfusion scans, and intraoperative inspection of the lungs. Linear regression analysis was performed on each patient using all serial postbronchodilator FEV1 values from before and after LVRS. RESULTS: Lung function data were available between 2-1001 days prior to LVRS and 71-1169 days after LVRS. Comparison of single pre- and post-LVRS FEV1 results confirmed a significant post-operative (3 month) improvement in lung function. The calculated rate of decline in FEV1 prior to LVRS was 202+/-205 mL/yr. Following LVRS, the rate of decline in FEV1 was unchanged at 178+/-150 mL/yr (p = 0.64). CONCLUSIONS: In patients with severe emphysema, bilateral LVRS does not appear to significantly alter the rate of FEV1 decline. PMID- 17136947 TI - Development of a quantifiable symptom assessment tool for patients with chronic bronchitis: the Chronic Bronchitis Symptoms Assessment Scale. AB - Clinical trials that rely solely on physiologic outcomes may underestimate effects of interventions on patients' symptoms and well-being. We sought to develop and validate an instrument to reliably assess symptom severity and frequency in therapeutic trials for patients with chronic bronchitis. We performed a series of psychometric studies that determined instrument content and assessed validity and reliability. The final version of the Chronic Bronchitis Symptoms Assessment Scale (CBSAS) comprised 15 scored items (16 items total). The instrument demonstrated strong reliability with moderate to high item correlations with total scores (0.41-0.80) and an overall Cronbach's alpha of 0.91. As expected, scores were at the bottom of the range for patients without a history of previous lung disease, and no ceiling effect was seen in the most severely obstructed patients. The CBSAS demonstrated statistically significant and moderate to strong association with spirometry, (post bronchodilator, % pred FEV1, r = -0.42, % pred FVC, r = -0.50), SGRQ (total, r = 0.67) and SOBQ (r = 0.50). The Chronic Bronchitis Symptoms Assessment Scale is an easy to administer, once daily or once weekly, valid and reliable outcome measure for assessing symptom severity among patients with chronic bronchitis. PMID- 17136948 TI - Costs and outcomes of extended-release vs. immediate-release clarithromycin for lower respiratory tract infections. AB - We used decision-analysis modeling to compare costs and outcomes of clarithromycin extended-release (Biaxin XL, Abbott Laboratories, Abbott Park, Illinois, USA) and clarithromycin immediate-release (Biaxin, Abbott Laboratories, Abbott Park, Illinois, USA) for outpatients with lower respiratory tract infections (LRTI). More patients achieved clinical cure with extended-release (83.9%) versus immediate-release (72.8%); fewer discontinued due to adverse events. Total costs with extended-release were $32 (16%) less; incorporating greater adherence for extended-release (once-daily) therapy resulted in greater savings. Results indicate that clarithromycin extended-release is cost-saving compared with immediate-release for LRTI. PMID- 17136949 TI - Development of a population-based screening questionnaire for COPD. AB - COPD is commonly under-diagnosed, in part because people at risk are unaware of the relevant risk factors and do not recognize related symptoms. Providing this information might permit earlier disease identification but the questions chosen should identify those with spirometrically defined airflow obstruction. Using a population-based data set, we have determined which questions identify persons most likely to have airflow obstruction. Potential questions were selected by review of COPD risk factors and clinical features. Validation was by retrospective analysis of the NHANES III data set, a population-based U.S. household survey that included spirometry. We examined the predictive ability of individual questions in a multi-variate framework to correctly discriminate between persons with and without spirometric airway obstruction (defined as FEV1/FVC < 0.70). We then tested the discriminatory ability of the questions in combination. The following items showed significant predictive ability: increased age, smoking status, pack-years, cough, wheeze, and prior diagnosis of asthma or COPD. The best performing combination was age, smoking status, pack-years smoked, wheeze, phlegm, body mass index, and prior diagnosis of obstructive lung disease. Using this combination in a population of current and former smokers aged 40 and over, we achieved a sensitivity of 85% and specificity of 45%, with a positive predictive value of 38% and a negative predictive value of 88%. Performance of this tool is comparable to other screening methods designed for use in a general population. Symptom-based questionnaires can be a viable method to identify persons likely to have COPD in the general population. Dissemination of such tools should raise awareness among at-risk persons and help identify COPD patients in the primary care setting. PMID- 17136950 TI - Effect of treating depression on quality-of-life and exercise tolerance in severe COPD. AB - The aim of this study was to determine whether treating concomitant depression improves quality of life and exercise tolerance in COPD patients. Out-patients with moderate to severe, stable COPD completed Hospital Anxiety-Depression (HAD) and General Health questionnaires. A psychiatrist interviewed those with high scores. In a randomised, double-blind fashion, 28 depressed COPD patients took a selective serotonin re-uptake inhibitor, Paroxetine 20 mg daily, or matched placebo for 6 weeks. Subsequently, all patients took un-blinded Paroxetine for 3 months. From these questionnaires, 35% of 135 patients had significant depression, but this was confirmed by psychiatric interview in only 21%. Throughout the study, there were no changes in laboratory lung function nor in home peak flow. Six weeks' treatment produced no significant differences between placebo and treatment group in either depression, quality of life scores or 6 minute walking distances, although overall improvements in depression, correlated with increases in walking distance. Three months of un-blinded treatment, significantly improved depression scores (self-complete HAD, Beck's Depression and psychiatrist-completed Montgomery-Asberg scores), walking distances (369 to 427 m, p = 0.0003) and St. George's Respiratory Questionnaire Total Scores (65 to 58, p = 0.033). Although self-complete questionnaires over-diagnose depression, the condition is nevertheless common in patients with moderately severe COPD. Six weeks of antidepressants is insufficient to improve either depression, quality of life or exercise tolerance. However, our study suggests that a longer course of treatment may be effective and that improvements in depression are associated with improvements in exercise tolerance. A larger, double blind study with a longer treatment period is indicated. PMID- 17136951 TI - Sleep disordered breathing in patients with chronic obstructive pulmonary disease. AB - Sleep-related disordered breathing (SDB) and its influence on desaturation were examined in stable COPD patients with waking SpO2 > 90%. With respiratory inductance plethysmography, thoracic-abdominal respiratory movements for all events with more than 4% desaturation were analyzed in 26 patients. Types of SDB were confirmed by full polysomnography. Irregular breathing induced desaturation, while stable respiration continued during some desaturation events. Three types of altered ventilation were observed: hypoventilation, paradoxical movement and periodic breathing. An unusual type of paradoxical movement, with normal airflow despite progressive desaturation, was observed in REM sleep. Patients were divided into desaturation (15 patients) and non-desaturation (11 patients) groups. Daytime arterial blood gas, lung function values, and 6-min walking distance did not differ. Awake, mode, maximum and minimum nocturnal SpO2 were lower in the desaturation group. SDB-induced desaturation events in the desaturation group were more frequent (9.2+/-3.5 vs. 1.8+/-2.2 times), a greater SpO2 decrease (11.4+/-7.1% vs. 5.2+/-2.1%) and longer duration (73.2+/-34.8 vs. 18.8+/-39.0 min). Patterns of SDB in the desaturation group were hypoventilation (74.4+/-23.4%), paradoxical movement (10.2+/-14.5%), periodic breathing (12.1+/ 18.3%) and unclassified (5.8+/-11.2%). These results reveal that lower SpO2 and SDB influence nocturnal desaturation in stable COPD patients. PMID- 17136952 TI - The airway pathophysiology of COPD: implications for treatment. AB - The pathophysiology of chronic obstructive pulmonary disease (COPD) is complex and can be attributed to multiple components: mucociliary dysfunction, airway inflammation and structural changes, all contributing to the development of airflow limitation, as well as an important systemic component. Current pharmacotherapies vary in their ability to address the underlying multi-component nature of COPD. Long-acting anticholinergics and long-acting beta2-agonists (LABAs) can both provide effective and convenient bronchodilation in moderate COPD (Stage II-GOLD) and are recommended as regular therapy in global treatment guidelines. However, there is evidence to suggest that LABAs can mediate additional benefits independent of their bronchodilatory effects and may help address the multi-component nature of COPD. Effects on mucociliary dysfunction and reduced bacterial-induced damage have been experimentally proven with LABAs, and anti-inflammatory activity and structural effects have also been suggested. The use of inhaled corticosteroids (ICSs) is now recommended for the treatment of COPD patients with frequent exacerbations. In addition, ICSs provide a range of anti-inflammatory effects in COPD and thus have effects that are complementary to those of LABAs. Recent data indicate that LABA/ICS combinations produce wide ranging clinical benefits that are greater than with either agent alone. Other new strategies include selective phosphodiesterase 4 (PDE4) inhibitors, which in addition to anti-inflammatory activity, have been shown to provide bronchodilation in COPD. In summary, the potential to address the multicomponent nature of COPD with strategies such as LABA/ICS combination therapy, and the development of new treatments directed at novel targets means that the future for sufferers of COPD can be more optimistic. PMID- 17136953 TI - A review of alpha-1 antitrypsin deficiency. AB - Alpha-1 antitrypsin (AAT) deficiency is a common but under-recognized disease. This hereditary disorder is characterized by low levels of AAT, and increased risks of panacinar emphysema at an early age, liver disease, vasculitis and panniculitis. Destruction of lung parenchyma and consequent emphysema result from an imbalance between different inflammatory proteases (in particular, leukocyte elastase), and the major natural antiprotease, AAT. To offer a review of key aspects of this important condition, we present the epidemiology, natural history, and pathogenesis of AAT deficiency and, in the context of recent data and the publication of an international standards document regarding the diagnosis and management of individuals with AAT deficiency, review current therapy of AAT deficiency. PMID- 17136954 TI - The Burden of Obstructive Lung Disease Initiative (BOLD): rationale and design. AB - Quantifying COPD prevalence worldwide is needed to document COPD's effect on disability, health care costs, and impaired quality of life and to inform governments and health planners. As an adjunct to data obtained from population based studies, and for countries where a fully powered prevalence survey cannot be done, modeling of COPD prevalence and its economic burdens can help estimate potential health care needs and costs. For comparability, standardized methods for prevalence surveys are needed that can be used in countries at all levels of economic development. The Burden of Obstructive Lung Disease (BOLD) Initiative has developed a set of methods for estimating COPD prevalence and a model for assessing its economic impact, and piloted these methods in China and Turkey. The methods were revised to reflect the findings in the pilot studies, and BOLD is now making the standardized methods available worldwide. The BOLD Operations Center provides training, materials, quality control, and data analysis. BOLD emphasizes data quality control at every stage of the process. Data from paper forms completed in the field are entered electronically to a specially designed secure Web platform. Pre- and post-bronchodilator spirometry testing is done on all participants, and all spirometry data are reviewed for quality. Questionnaires are used to obtain information about respiratory symptoms, health status, exposure to risk factors, and economic data about the burden of COPD. BOLD's standardized methods will provide a uniform way to compare COPD burden within and between countries, and where differences are found, to explore explanations for these differences. PMID- 17136955 TI - Interpreting results from clinical trials: understanding minimal clinically important differences in COPD outcomes. PMID- 17136956 TI - A two-stage logistic model based on the measurement of pro-inflammatory cytokines in bronchial secretions for assessing bacterial, viral, and non-infectious origin of COPD exacerbations. AB - Exacerbations often complicate the progressive course of chronic obstructive pulmonary disease (COPD), mainly due to infectious agents. The precise role of bacterial infections in the course and the pathogenesis of COPD has been a source of controversy for decades. Also viruses and other non-infectious causes of exacerbation play a relevant role and also contribute to persisting airway inflammation. Usually, the etiologic identification of the infective causes of COPD require considerable time and costs. The development of more rapid, reliable, and widely applicable methods to promptly define the etiology of COPD exacerbations should represent a relevant issue in devising earlier and more specific strategies for their effective therapeutic control. AIM: Of the study was to assess the predictive role of some pro-inflammatory cytokines measured in spontaneous bronchial secretions in discriminating the main infectious causes of COPD exacerbations. METHODS: 124 subjects with moderate COPD (51-79 y; mean basal FEV1 = 49.6% pred. +/- 4.6 sd; FEV1 reversibility +3.9% from baseline +/- 4.8 sd after salbutamol 200 mcg) were studied during acute exacerbation. Respiratory viruses were isolated from bronchial secretions in 21 cases; common bacteria (CFU > or = 10(6)/ml) in 28 cases; Pseudomonas Aeruginosa (Ps.Ae.; CFU > or = 10(6)/ml) in 20 cases. The cytokines IL1beta, IL8, and TNFalpha (pg/ml; Immulite; Diagnostic Product Corp, Los Angeles, CA, USA), and neutrophils (% total count) were measured in bronchial secretions of all patients. STATISTICS: A two-stage logistic model was chosen for discriminating the different causes of COPD exacerbations (such as: non-infectious, or viral, bacterial, or due to Ps.Ae.). RESULTS: At the first decisional step, the two-stage logistic model proved that TNFalpha levels in bronchial secretions recognise clearly patients belonging to the Ps.Ae. group from those of all other groups (Area under ROC curve = 0.96; 95% CI = 0.91-0.99), and that, at the second decisional step, IL8 + IL10 levels discriminate patients with bacterial causes (such as all bacteria) from the non infected ones and from those with a viral cause of exacerbation (Area under ROC curve = 0.87; 95% CI = 0.77-0.94). Neutrophil percent count did not support any contribution in discriminating the different subgroups of COPD subjects. CONCLUSIONS: When exacerbated, COPD subjects express different patterns of pro inflammatory mediators in bronchial secretions, which appear modulated according to the etiological cause of the exacerbation. In particular, TNFalpha concentration per se enables recognition of COPD exacerbations due to Ps.Ae., while IL8 + IL1beta levels prove helpful in discriminating those to common bacteria from those to viral agents and to non-infectious causes. When present data are further confirmed, the use of a decisional rule based on cytokine measurements might be regarded as a helpful predictive tool. As measures of pro inflammatory cytokines are low-cost, simple, and faster to perform, they could support rapid clinical decision making at the bedside regarding therapeutic strategy for COPD exacerbations, in particular when they are needed for severe COPD patients. PMID- 17136957 TI - Different modulation of decorin production by lung fibroblasts from patients with mild and severe emphysema. AB - We have previously reported diminished immunohistochemical staining of decorin in lung tissue from patients with severe emphysema. The aim of this study is to investigate whether this diminished staining is due to a quantitative abnormal production of decorin by pulmonary fibroblasts in vitro. Therefore, we measured decorin (Western blot), collagen type I (ELISA), and fibronectin (ELISA) production by fibroblasts obtained from lung tissue of patients with severe and mild emphysema at basal culture conditions and after modulation with transforming growth factor-beta1, basic fibroblast growth factor, and interferon-gamma. Decorin production at basal culture conditions was significantly higher in fibroblast cultures from patients with severe emphysema compared to fibroblasts from mild emphysema. After stimulation with transforming growth factor-beta1 and basic fibroblast growth factor, decorin production was significantly more reduced in fibroblast cultures from patients with severe emphysema whereas collagen type I and fibronectin production were not affected. We conclude that decorin production by lung fibroblasts of patients with severe emphysema is dysregulated after modulation with cytokines known to be important in smoking associated inflammation. This dysregulation of decorin production may contribute to the impaired lung tissue repair, present in patients with emphysema, since these alterations in the extracellular matrix may cause diminished cytokine binding and neutralization. PMID- 17136958 TI - Outcome in adult bronchiectasis. AB - The outcome in adult bronchiectasis has not been well described; in particular there has been a lack of long-term prospective studies. Therefore a follow-up study was performed to assess outcome in bronchiectasis in a cohort of adult patients. One hundred-and-one sequential adults, 33 male and 68 female; age 54 +/ 14 years (mean +/- SD) with bronchiectasis had a clinical assessment and spirometry performed. All were non-smokers and 84 were classified as having idiopathic disease. Patients were commenced on a standardized treatment regime and followed up for a minimum period of 2 years. On their last review when patients were clinically stable, a repeat clinical assessment and spirometry was performed and compared with the initial review. The primary endpoints measured were symptoms and FEV1. Subjects were followed up for 8.0 +/- 4.9 years. Clinical review showed that the patients had persistent symptoms that, in the case of dyspnea and sputum volume, were worse on follow-up. Spirometry showed a significant decline in FEV1 over the follow-up period with an average loss of 49 ml per year. This study showed in this group of predominantly female adult patients with bronchiectasis followed up for 8 years, patients had persistent symptoms and an excess loss in FEV1. PMID- 17136959 TI - Trends and cardiovascular co-morbidities of COPD patients in the Veterans Administration Medical System, 1991-1999. AB - Persons with Chronic Obstructive Pulmonary Disease (COPD) are at risk for developing other smoking-related complications, including cardiac and vascular diseases. Information about the prevalence and incidence of these conditions is needed to anticipate their occurrence in clinical research. We conducted a cohort study using longitudinal administrative data to describe the prevalence and incidence of cardiovascular (CV) diseases among COPD patients treated by the Veterans Administration Medical System. The COPD cohort included all persons with a diagnosis of COPD admitted to a Veterans Administration Medical System hospital (N = 70,679) or seen in a outpatient clinic (N = 314,209) in fiscal year 1998. Each COPD patient was matched to a Veteran of the same age and gender who did not have a COPD diagnosis, creating a non-COPD cohort for comparison. Among all hospitalized Veterans, the prevalence of COPD steadily increased from 1991 to 1999, although the total number of Veterans hospitalized during this time period decreased by more than one-third. Among COPD patients hospitalized in 1998, the prevalence of coronary artery disease, congestive heart failure, and atrial fibrillation were very high (33.6%, 24.4%, and 14.3%, respectively) and significantly higher than those seen among the matched non-COPD cohort (27.1%, 13.5%, and 10.4%; p < 0.001). Among COPD outpatients, increased complications were found in every CV disease category with rate ratios that were greater than observed among inpatients. We conclude that CV diseases are remarkably prevalent among Veterans with COPD, and their incidence is likely to increase as the Veteran population ages. PMID- 17136960 TI - Challenges associated with estimating minimal clinically important differences in COPD-the NHLBI perspective. AB - Chronic Obstructive Pulmonary Disease (COPD) is a common lung disease that exemplifies the value, as well as the difficulties and challenges, of using minimal clinically important differences (MCID) in clinical research. Development and validation of better endpoints for clinical studies is critical to research progress in COPD. However, the clinical, genetic, and pharmacological heterogeneity of the COPD patient population complicates attempts to define and validate MCIDs for COPD. It is difficult to identify a single measurable outcome that reflects the many components of the COPD patient's health state. Acute exacerbations of symptoms, which COPD patients often experience, present another challenge in the development of MCIDs for this disease. Consequently, the NHLBI does not require the use of MCIDs in clinical research. This allows research on the causes, prevention and diagnosis of COPD and use of endpoints for which an MCID is not yet known. It is important for the scientific community to reach agreement on what is a meaningful MCID in therapeutic trials for COPD. Further research into the concept of the MCID and its application should enable therapeutic trials in COPD to yield knowledge that is more effectively translated into improved public health. PMID- 17136961 TI - U.S. regulatory perspective on the minimal clinically important difference in chronic obstructive pulmonary disease. AB - This paper outlines the regulatory issues surrounding the determination and use of minimally clinically important differences (MCID) in assessing measures of outcomes from treatments of chronic obstructive pulmonary disease (COPD). To place this discussion in context, it is important to understand the current state of approved therapies for COPD, as well as newer directions in therapy. This paper discusses the currently available, approved drug therapies for COPD in the United States and how they were approved. This is followed by an overview on the use of MCID for assessing outcomes in therapies for COPD, as well the more general experience with MCID from the U.S. regulatory perspective. PMID- 17136962 TI - Minimal clinically important difference, clinical perspective: an opinion. AB - The Minimal Clinically Important Difference has become a key feature for both the validation of clinical tools and for the assessment of clinical studies. Several methods have been developed to establish what a Minimal Clinically Important Difference is. The primary purpose of the Minimal Clinically Important Difference, however, is to provide a measure of relevance for a statistically applied measure. It does not, despite its name, necessarily relate to the "Clinical" condition. In this context, human beings are capable of extremely fine grades of discrimination of very subtle differences, when they care about the measures. When they do not care about the measure, large differences may be irrelevant. The flavors of wines or the tone qualities of musical instruments are readily recognizable examples. The importance of an outcome, for a clinician caring for an individual patient, therefore, will be highly patient specific. The Minimal Clinically Important Difference has great utility in assessing tools for clinical investigation. It has limitations in assisting the clinician. The subtleties that may be meaningful to individuals are often lost in discrimination tests in large populations where not all have the same interests. In addition, readily applicable tests, for example, discriminating degrees of salty water, for which Minimal Clinically Important Difference can be readily defined, are often tests that have no interest for the majority of the population tested. This leads to several paradoxes. Readily defined Minimal Clinically Important Differences are likely to be defined for parameters that are of little interest to a large number of persons. Conversely, parameters that are of great interest to selected individuals, that could be discerned by them with great subtly are likely to be poorly generalizable. Without doubt, defining a Minimal Clinically Important Difference will remain a key goal in the validation and application of tools for clinical investigations. The limits of the concept, particularly as it relates to issues of importance to patients, however, needs to be recognized. PMID- 17136963 TI - Assessing the minimally clinically significant difference: scientific considerations, challenges and solutions. AB - The scientific considerations surrounding the estimation of a minimally clinically important difference (MCID) are a myriad and challenging. There are a considerable number of hurdles to overcome. The good news is that there are solutions to virtually every one of the scientific hurdles. This paper intends to set out the issues, identify the challenges, and offer solutions so that the state of the science may move forward. The ultimate outcome of the paper may not be to provide a definitive answer for estimating the MCID in every situation, but it should provide a starting point and advice for a process or set of guidelines that may be followed toward achieving this goal. The paper begins with a brief synthesis of the literature and state of the science at the time of publication. The relationship between the process for determining MCIDs for other endpoints, such as tumor response or complete blood culture (CBC) variables, versus toxicity and QOL-related variables is described. The ultimate lessons to be learned from this exercise are: 1. There are many methods available to ascertaining an MCID. None are perfect, but all are useful. 2. All methods converge to similar answers. Supplementary information may refine answers from one or more of the methods. 3. Clinical opinion and patient subjective response should trump statistical theory. 4. A process of MCID estimation involving all approaches to produce a potential range with sensitivity analyses is the optimal solution to producing an MCID based on the most complete knowledge possible. PMID- 17136964 TI - Approaches and recommendations for estimating minimally important differences for health-related quality of life measures. AB - We describe currently available approaches for estimating the minimally important difference (MID) and their associated strengths and weaknesses. Specifically, we show that anchor-based methods should be the primary method of estimating the MID because of the limitations of distribution-based methods. In addition, we provide recommendations for estimating the MID in future research. PMID- 17136965 TI - The relation between the minimally important difference and patient benefit. AB - A critical issue in the examination of the effects of treatments on health related quality of life is how to determine whether a particular change is clinically relevant. One approach is the so-called anchor-based method derived from patient or clinician estimates of minimal change (the Minimally Important Difference or MID). At issue, however, is whether this criterion provides a meaningful way to differentiate between beneficial and ineffective treatments. In this paper, I show that the likelihood that a patient will benefit from treatment, or alternatively, the number of patients in a given cohort who will benefit from treatment, can be predicted with considerable precision from the Effect Size, and the particular choice of MID bears almost no relation to the projected benefit. To examine the relation between the threshold of minimal difference, the effect size of treatment, and the likelihood that a patient will benefit from treatment, a simulation based on a normal distribution was used to compute the proportion of patients benefiting for various values of the ES and the MID. The agreement of the simulation with empirical data from four studies of asthma and respiratory disease was examined. The simulation showed a near-linear relationship between ES and the likelihood of benefit, which was nearly independent of the value of the MID. Agreement of the simulation with the empirical data was excellent. Introducing moderate skew into the distributions had minimal impact on the relationship. The proportion of patients who will benefit from treatment can be directly estimated from the effect size, and is nearly independent of the choice of MID. Effect size- and anchor-based approaches provide equivalent information in this situation. There appears to be little utility in the notion of the MID as an absolute indicator of clinically important treatment effects. PMID- 17136966 TI - St. George's Respiratory Questionnaire: MCID. AB - The SGRQ is a disease-specific measure of health status for use in COPD. A number of methods have been used for estimating its minimum clinically important difference (MCID). These include both expert and patient preference-based estimates. Anchor-based methods have also been used. The calculated MCID from those studies was consistently around 4 units, regardless of assessment method. By contrast, the MCID calculated using distribution-based methods varied across studies and permitted no consistent estimate. All measurements of clinical significance contain sample and measurement error. They also require value judgements, if not about the calculation of the MCID itself then about the anchors used to estimate it. Under these circumstances, greater weight should be placed upon the overall body of evidence for an MCID, rather than one single method. For that reason, estimates of MCID should be used as indicative values. Methods of analysing clinical trial results should reflect this, and use appropriate statistical tests for comparison with the MCID. Treatments for COPD that produced an improvement in SGRQ of the order of 4 units in clinical trials have subsequently found wide acceptance once in clinical practice, so it seems reasonable to expect any new treatment proposed for COPD to produce an advantage over placebo that is not significantly inferior to a 4-unit difference. PMID- 17136967 TI - Measurement properties and interpretability of the Chronic respiratory disease questionnaire (CRQ). AB - The chronic respiratory questionnaire, available as an interviewer and a self administered instrument, includes 20 items across four domains: dyspnea (5 items), fatigue (4 items), emotional function (7 items), and mastery (4 items). When completing this instrument, patients rate their experience on a 7-point scale ranging from 1 (maximum impairment) to 7 (no impairment). The Chronic Respiratory Questionnaire has demonstrated excellent measurement properties for both discriminative and evaluative purposes and served as a model in numerous methodological studies in chronic airflow limitation and patients with chronic obstructive pulmonary disease. We performed a systematic review of the literature on the chronic respiratory questionnaire to summarize the key qualities of the chronic respiratory questionnaire and to appraise the work regarding the minimal important difference of the chronic respiratory questionnaire. This paper includes a revision of our initial definition of the minimal important difference and a methodological framework for using anchor based approaches to establish the minimal important difference pioneered by Jaeschke and colleagues. Other approaches to evaluate the minimal important difference include distribution based methods and panel-based methods. Investigators have used all of these approaches to establish the minimal important difference for the chronic respiratory questionnaire and the results are in general agreement with the minimal important difference of 0.5 for the mean domain scores of the chronic respiratory questionnaire. As a result of this literature review and discussion at the workshop, we established several research objectives. These objectives include the exploration of presentation of quality of life information and prospective anchor-based approaches. PMID- 17136968 TI - The minimally clinically important difference in generic utility-based measures. AB - PURPOSE: To evaluate the use of utility-based generic quality of life measures for establishing the minimally clinically important difference (MCID). BACKGROUND: Utility-based quality of life measures place levels of wellness on a continuum anchored by death (0.00) and optimum function (1.00). Preference measurement studies are used to define the meaning of points along the continuum. Health states that differ by less than 0.03 units cannot be discriminated by panels of judges as different from one another. Thus, 0.03 is a reasonable MCID for these measures. METHOD: Three published studies of patients with Chronic Obstructive Pulmonary Disease (COPD) reported data on the Quality of Well-being Scale (QWB) before and after pulmonary rehabilitation. One of the studies also randomly assigned patients to lung volume reduction surgery or to maximal medical therapy. These patients were followed for an average of 29 months. RESULTS: All three evaluations of pulmonary rehabilitation showed changes on the QWB in excess of the proposed 0.03 MCID. QWB changes for patients assigned to lung volume reduction surgery were close to the MCID threshold at one year but grew stronger in subsequent years. Using Norman's 0.50 standard deviation method, all three estimates of rehabilitation effectiveness and the outcomes one year following surgery fall below the MCID. CONCLUSION: Different methods for estimating MCID lead to different conclusions about the meaning of quality of life changes following pulmonary rehabilitation and lung volume reduction surgery. The preference scaling system in generic utility-based quality of life measures provides a metric that is directly interpretable and avoids many of the criticisms of MCID measures. The method is sensitive enough to suggest clinically meaningful benefits of rehabilitation and surgery. Further, quality adjusted life years offer a valuable metric for policy analysis. Utility-based measures of health related quality of life should gain greater use in COPD outcomes research. PMID- 17136969 TI - The MCID of the transition dyspnea index is a total score of one unit. AB - The Baseline (BDI) and Transition (TDI) Dyspnea Indexes provide interview-based measurements of breathlessness related to activities of daily living. The BDI is a discriminative instrument that includes specific criteria for each of three components at a single point in time. The TDI is an evaluative instrument that includes specific criteria for each of three components to measure changes from a baseline state. Observational studies have shown that patients with COPD generally experience a gradual progression of breathing difficulty as measured by the TDI over time. Randomized controlled trials have demonstrated excellent measurement characteristics of the TDI; these include responsiveness (ability to detect change) and construct validity (a change in the TDI correlates with changes in other variables). Supporting evidence for one unit as the minimal clinically important difference (MCID) of the TDI is based on: expert preference; use of the physician's global evaluation score as an anchor; and distribution estimates (standard error of measurement and 0.5 of the standard deviation). As an alternative to the interview process, self-administered computerized (SAC) versions of the BDI/TDI have been developed to provide direct patient-reported ratings of dyspnea. To further establish the MCID of the interview-administered and/or the SAC TDI, we recommend that a patient's report of global ratings of change by used as an independent standard or anchor. PMID- 17136970 TI - Minimally clinically important difference for the UCSD Shortness of Breath Questionnaire, Borg Scale, and Visual Analog Scale. AB - Dyspnea is a primary symptom of chronic lung disease and an important outcome measure for clinical trials. Several standardized measures have been developed to evaluate this important symptom and are being used increasingly in clinical trials. The minimally clinically important difference (MCID) is not well defined for these measures but is important in interpreting the clinical meaning of results of studies in this area. The purpose of this paper is to evaluate the MCID for three commonly used measures to assess dyspnea in chronic lung disease: UCSD Shortness of Breath Questionnaire (SOBQ), Borg Scale (Borg), and Visual Analog Scale (VAS). The analysis is based on a retrospective review of published trials evaluating the response to a pulmonary rehabilitation or exercise intervention that is known to produce modest, but clinically meaningful changes for such patients. Using a distribution-based approach based primarily on effect size, the recommended MCID for these measures are: 5-units for the SOBQ, 1-unit for the Borg scale, and approximately 10 to 20 units for the VAS. PMID- 17136971 TI - Minimal clinically important differences in COPD lung function. AB - The FEV1 is widely used by physicians in the diagnosis, staging, treatment, monitoring, and establishing prognosis for patients with COPD. The MCID is the smallest difference which patients perceive as beneficial and which would mandate a change in patient management. A precise MCID for FEV1 has not been established. In attempt to establish a MCID for predose or trough FEV1, several limitations need to be addressed. There are issues such as reproducibility, repeatability, acceptability, variability, placebo effect, and equipment effects. Patient factors, such as baseline level of FEV1, albuterol reversibility, diurnal variation, influence the results. Nonetheless, using anchoring techniques, a change in pre dose FEV1 of about 100 mL can be perceived by patients, correlates with fewer relapses following exacerbations and is in the range usually achieved with bronchodilators approved for COPD. In the future, consistent reporting of spirometric variables, such as a predose FEV1 and other outcomes, can be incorporated into a more quantitative effort to establish the MCID. Also distributional/statistical methods may be useful in determining the MCID FEV1. PMID- 17136972 TI - Minimal clinically important differences in the six-minute walk test and the incremental shuttle walking test. AB - Simple walking tests are widely used for the assessment of functional status in patients with cardiorespiratory disorders. These tests require far less instrumentation than formal cardiopulmonary exercise tests, but they do require standardization of procedures to achieve reproducible results. The most widely used tests for patients with COPD are the 6-minute walking test (6MWT) and the incremental shuttle walking test (SWT). The 6MWT has been characterized in COPD patients with respect to reproducibility and responsivity to change in health status. The 6MWT results are correlated with pulmonary function, health-related quality of life, maximum exercise capacity, and mortality. The minimal clinically important difference (MCID) for the 6MWT is conservatively estimated to be 54-80 meters using both distributional and discriminative methods. For an individual patient, the 6MWT would need to change by about 86 meters to be statistically confident that there has been a change. The SWT has been less extensively validated than the 6MWT, but has similar reproducibility in COPD (CV = approximately 20%). The SWT results improve with pulmonary rehabilitation and bronchodilation, and are highly correlated with maximum oxygen consumption. There are no studies that address the issue of MCID for the SWT. In addition to the MCID, the design and interpretation of COPD clinical trials should take into account the severity of initial impairment, the asymmetry between positive and negative changes, the proportion of patients who show substantial improvement, and the costs and risks of the treatment. PMID- 17136973 TI - Factors determining constant work rate exercise tolerance in COPD and their role in dictating the minimal clinically important difference in response to interventions. AB - Constant work rate exercise testing has been used to assess effectiveness of therapeutic interventions in chronic obstructive pulmonary disease. It has been noted to yield larger fractional increases in exercise tolerance than other measures. Reasons for this are rooted in physiological determinants of the time course of pulmonary ventilation. Following exercise onset, ventilation increases in three phases; the slowest phase is seen only at high exercise intensities. In chronic obstructive pulmonary disease exercise proceeds until ventilation reaches a limiting value. Because both ventilatory requirement for exercise and limiting ventilation can be manipulated by several interventions (exercise training, oxygen inhalation, bronchodilator administration), constant work rate tolerance responds to these interventions. The power-duration relationship predicts the effect of the work rate imposed on tolerated duration of a constant work rate test. Arguments are presented that a pre-intervention constant work rate duration of 4-7 minutes is desirable. At present, the recommended strategy to achieve this target duration is to choose a work rate equal to 85% of peak work rate achieved in a constant work rate test. There is clearly insufficient information to reliably determine a minimal clinically important difference for this test. A lower bound of 1.75 minutes is suggested. PMID- 17136974 TI - Maximum exercise as an outcome in COPD: minimal clinically important difference. AB - Limitation of physical activity occupies a central role in the symptom complex of patients with chronic obstructive pulmonary disease (COPD), and improvement in exercise capacity is a key outcome of response to COPD therapy. Maximum exercise capacity testing facilitates assessment of physiologic mechanisms of exercise and allows quantitation of the degree of limitation. This manuscript utilizes published data from the National Emphysema Treatment Trial to investigate the minimal clinically important difference (MCID) in maximum exercise capacity in patients with severe emphysema. Distribution- and opinion-based methods were used to estimate MCID. Expert clinician opinion yielded a value of 10 Watts as the MCID for change in maximum exercise capacity. Baseline standard deviation and error data yielded a one-half standard deviation-based estimate of 10.5 Watts and a standard error-based estimate of 4.2 Watts. In subjects randomized to medical therapy, the mean (+/-SD) 24-month change in maximum exercise capacity following medical therapy was -9.2 +/- 1.2 Watts, whereas among those randomized to lung volume reduction surgery, mean 24-month change in maximum exercise capacity was 1.7 +/- 17.7 Watts, with a mean difference between the groups of 10.9 Watts. The observed difference in maximum exercise capacity after 24 months between subjects randomized to medical versus surgical therapy conforms to both opinion- and distribution-based estimates of MCID. Further investigation is needed to develop and validate estimates of MCID for maximum exercise capacity and other key clinical outcomes in COPD. PMID- 17136975 TI - Minimal clinically important difference--exacerbations of COPD. AB - Exacerbations of COPD are now recognised as being important events in the natural history of the condition and become more frequent as the disease worsens. Defining a minimum clinically important difference in exacerbation rate is fraught with difficulty. There is substantial between and within subject differences in the occurrence of these events that makes an individual evaluation of their importance problematic. At present, the most widely used definition of an exacerbation identifies an episode where the patient seeks medical help rather than a predefined change in one or more symptoms. Despite these problems, intervention studies with bronchodilator drugs, inhaled corticosteroids, and pulmonary rehabilitation appear to reduce the frequency of exacerbation events. In patients with an FEV1 below 50% predicted there is reasonable consistency about the magnitude of change and a 4-unit improvement in the St George's Respiratory Questionnaire is commonly associated with a 20-25% reduction in the reported number of exacerbations. Individual studies vary depending upon the recruitment protocol. Patients who experience symptomatic benefit may be those in whom a clinically important change in exacerbations occurs but this concept requires testing prospectively. Existing methodologies for estimating clinically important differences are hard to apply with a binary outcome like this, and more work will be needed to develop a robust approach for dealing with this important clinical variable. PMID- 17136976 TI - Minimal clinically important differences in COPD: body mass index and muscle strength. AB - Weight loss, muscle wasting, as well as muscle dysfunction are recognized as important problems in COPD, contributing to morbidity and mortality. This paper discusses body weight and muscle function as possible outcome parameters in the management of COPD. The relationship between these outcome measures and COPD related management goals is discussed. Minimal clinically important differences (MCID) in the approach of patients suffering from COPD for these measures are discussed. PMID- 17136977 TI - Bridging the gap: using triangulation methodology to estimate minimal clinically important differences (MCIDs). AB - This paper proposes the use of triangulation methodology to derive guidelines for interpreting change scores on health outcome measures. Triangulation integrates results from global ratings with clinical benchmarks of change, statistical estimates of magnitude, and qualitative data from patients and/or clinicians to derive guidelines that are not field-specific or method bound. A case study is presented to illustrate how this methodology can be applied. Secondary analyses were performed on blinded data from 2,971 patients enrolled in three phase IlIa clinical trials to develop guidelines for interpreting change scores on the Breathlessness Diary (BD), a relatively new approach for evaluating dyspnea outcomes in patients with chronic obstructive pulmonary disease. BD scores were examined by disease severity and rescue medication use. In addition, mean BD change scores by physician global ratings of efficacy were juxtaposed with changes in forced expiratory volume (FEV1) and St. George's Respiratory Questionnaire scores. Percent change, effect size, one-half standard deviation, and the standard error of measurement were used as statistical indicators of magnitude. Data from qualitative interviews provided insight into patient perspectives of change in dyspnea. Taking into consideration results across estimation methods, guidelines were developed for defining large, moderate, and small group-level mean changes on the BD. Areas of divergence and convergence across statistical indicators and clinical benchmarks in this case study highlight the importance of using triangulation methodology to derive guidelines that are both empirically sound and clinically relevant. PMID- 17136978 TI - Clinical efficacy of OM-85 BV in COPD and chronic bronchitis: a systematic review. AB - OM-85 BV is an immunomodulatory agent used for prevention of exacerbations in persons with chronic lung disease. We conducted a systematic review of OM-85 BV to evaluate its efficacy and safety. A systematic search for relevant articles was performed. Studies were included if they involved persons with chronic obstructive pulmonary disease or chronic bronchitis and were randomized to OM-85 BV or placebo. Investigators extracted data on study design, participant characteristics, and clinical outcomes. Thirteen trials involving 2066 individuals met inclusion criteria. Three trials enrolled an older, more homogenous population with chronic obstructive pulmonary disease. Utilizing quantitative pooled analysis in these studies, with one or more acute exacerbations as the endpoint, we found a non-statistically significant trend in favor of OM-85 BV [relative risk 0.83, 95% confidence interval 0.65-1.05]. Ten trials enrolled a heterogeneous population with chronic bronchitis. In these trials, exacerbation rates were less with OM-85 BV in 4 of 9 trials reporting this outcome. Varied results in the outcomes of hospitalization, symptom scores, and antibiotic or steroid use were found across studies. Withdrawals and adverse events were similar between OM-85 BV and placebo. While OM-85 BV is used to prevent exacerbations in persons with chronic lung disease, consistent evidence across multiple important outcomes does not exist to clearly demonstrate clinical benefit. Further randomized controlled trials enrolling large numbers of persons with well-defined COPD are necessary to confirm the effectiveness of this agent. PMID- 17136979 TI - The challenges of chronic obstructive pulmonary diseases (COPD)--a perspective. AB - Chronic Obstructive Pulmonary Disease (COPD) is an inflammatory disease, primarily caused by cigarette smoke, which will soon become the third leading cause of death globally. Despite the importance of the problem, our real understanding of the biological underpinnings of COPD remains incomplete. Consequently, our first-line therapies, while helpful, are not yet as effective as they need to be. In this review, we will focus on these challenges and more, including the role of impaired tissue repair and adaptive immunity in disease pathogenesis, determining who may be at risk, describing COPD phenotypes and potential biomarkers. New ideas for chronic disease management and prevention of exacerbations will also be discussed. While much remains to be accomplished, meeting these challenges will bring rewards because what we learn will have implications for the understanding and treatment of chronic inflammatory diseases beyond COPD. PMID- 17136980 TI - Current perspectives on management of co-morbid depression in COPD. AB - Individuals with COPD have a higher prevalence of co-morbid depression than either the general population or patients with other chronic illnesses. The best estimates report a prevalence of approximately 40% in COPD patients, compared to 15% in the general population. Depression in COPD patients leads to a lower quality of life, greater objective impairment in function, and decreased adherence to therapeutic interventions. While many depressed COPD patients have been treated empirically with antidepressants--subjecting them to antidepressant side effects, toxicities, and costs--there is a surprising lack of evidence supporting or directing that treatment. We review the current literature regarding the management of depression in COPD, suggest strategies for management, and future research needs. PMID- 17136981 TI - Alphabet soup: assessment of dyspnea. PMID- 17136982 TI - Lung tissue banks for gene expression studies. PMID- 17136983 TI - Analysis of systemic biomarkers in COPD patients. AB - The finding that alphal-antitrypsin (AAT) deficiency, PiZZ, a well-established genetic risk factor for COPD, is related to high levels of circulating AAT polymers, prompted us to measure serum levels of such polymers and selected markers of inflammation in age- and gender-matched patients with stable COPD and control subjects with and without severe AAT deficiency, and to assess their relationship with each other and with the genetic AAT-variant. We found that COPD individuals (n= 20), independent of AAT-variant, had significantly higher serum levels of AAT and its polymers, MMP-9, sICAM-1, VEGF and sE-selectin than controls (n=30). Subjects with PiZZ COPD (n= 10) showed significantly elevated serum levels of AAT-polymers, sE-selectin and sICAM-1, while patients with PiMM COPD (n= 10) showed higher levels of MMP-9, VEGF, IL-8 and MCP-1 than controls. By using factor analysis we were able to split the analysed biomarkers into two independent components: the first containing MMP-9, MCP-1, IL-8 and VEGF and the second-AAT and its polymers and sE-selectin. The result from the binomial logistic regression showed that 95.2 percent of the control individuals and 94.7 percent of the COPD patients can be correctly classified on the basis of the measured serum biomarkers. These observations highlight the importance of the finding sets of biomolecules, which could offer new strategies for the diagnosis of COPD and may have value for monitoring progression of COPD. PMID- 17136984 TI - Development of self-administered versions of modified baseline and transition dyspnea indexes in COPD. AB - In this study we developed self-administered versions of modified baseline and transition dyspnea indexes and compared the scores obtained by this method with the mean value obtained by two trained interviewers. Twenty-five patients (14 males/11 females) with chronic obstructive disease who had a chief complaint of "breathlessness" were tested. Age was 66+/-11 years; forced expiratory volume in one second was 48+/-23% predicted. The baseline total scores were 5.0+/-1.8 for the interviewers and 5.4+/-2.0 for the self-administered method. For the baseline dyspnea scores the correlations were 0.83 (p<0.0001) between self-administration and the mean value of two interviewers and 0.75 (p<0.0001) between the two interviewers. The transition total scores, obtained an average of 102 days (range, 7-377 days) later, were - 0.1+/-3.0 for the interviewers and - 0.4+/-3.0 for the self-administered method. For the transition dyspnea scores the correlations were 0.94 (p<0.0001) between self-administration and the mean value of two interviewers and 0.83 (p<0.0001) between the two interviewers. The self administered dyspnea scores had similar correlations with measures of lung function as did the interview dyspnea scores. We conclude that self-administered versions of the modified baseline and transition dyspnea indexes provide comparable scores as those obtained by trained and experienced interviewers. The advantages of the self-administered versions include standardized methodology and computerized scoring. PMID- 17136985 TI - Prevalence and risk factors for chronic obstructive pulmonary disease according to symptoms and spirometry. AB - The burden of COPD is quite high and its prevalence is increasing. Few data are available from Latin America. There is no consensus on what criteria should be the gold standard for the definition of this disease. A population-based study was carried out in a southern Brazilian city, including adults aged 40-69 years. The aim was to measure the prevalence of COPD according to several criteria. From the 1,046 subjects chosen from a multiple-stage sampling protocol, a sub-sample of 234 subjects was systematically selected to undergo spirometry. Percentages of COPD according to different criteria were: chronic bronchitis by questionnaire (7.8%); GOLD stage 0 (7.3%); fixed ratio (15.2%); GOLD stage II (9.9%); ERS (27.7%). The relationship among the three lung functional measurements showed that around 70% of all subjects had negative results with all three criteria, and around 10% were positive according to all three. Utilization of symptom-based or spirometry definitions of COPD provide different prevalence estimates. Use of different spirometric criteria also resulted in different percentages of COPD. According to spirometry, COPD was higher among men, elderly, those with low education and ex-smokers. On the other hand, current smokers were more likely to present symptoms of chronic bronchitis. The fixed ratio criterion is recommended when population-specific reference curves are not available, while the GOLD definition is recommended to evaluate severity of COPD. PMID- 17136986 TI - Eletrocardiographic monitoring in COPD patients receiving tiotropium. AB - Tiotropium is a once-daily, inhaled anticholinergic for the treatment of chronic obstructive pulmonary disease that acts as a prolonged antagonist of the M3 receptor. To ascertain whether electrophysiologic effects can be detected following tiotropium treatment in patients with chronic obstructive pulmonary disease, serial electrocardiograms were incorporated into multiple placebo controlled clinical trials including long-term (6 and 12-month) trials with tiotropium 18 mcg daily (n=2,128) and a 4-week dose-ranging study with tiotropium up to 36 mcg daily (n= 169). In addition, 24-hour electrocardiographic (Holter) monitoring was performed as part of a 6-week, placebo-controlled trial with tiotropium 18 mcg daily (n= 121). Electrocardiograms were performed before and up to 6 times during treatment in the 12-month trials, and before and at the end of treatment in the 6-month trials. For both the 12 and 6-month trials, electrocardiograms were recorded as adverse events if significant changes occurred, and were retrospectively sent for centralized analysis. During the 6 week trial, Holter monitoring was performed prior to the first dose and following 6 weeks of treatment. In all of these trials, no significant differences were observed in any of the electrocardiogram or Holter outcome parameters compared to placebo. Specifically, there was no clinically relevant difference in heart rate, atrio-ventricular conduction or the occurrence of ventricular or supraventricular arrhythmias. In conclusion, tiotropium was not associated with any signs of cardiac safety concerns as defined by electrocardiographic evaluations in placebo controlled clinical trials. PMID- 17136987 TI - A lung tissue bank for gene expression studies in chronic obstructive pulmonary disease. AB - A bank of surgically resected human lung tissues frozen at -70 degrees C after being inflated with support medium for cutting frozen tissue and a separate group inflated with fixative and embedded in paraffin has been established for studies of chronic obstructive pulmonary disease. The present report concerns the quality of RNA that can be extracted from these frozen and fixed tissue samples and from cells obtained from them by laser capture microdissection. The results show that the RNA yield was 257+/-183 ng/mg and 77+/-56 ng/mg from randomly selected frozen and paraffin-embedded tissue, respectively. Intact 18S and 28S rRNA subunits were present in 11/23 frozen and 2/6 paraffin-embedded specimens. The 375-bp actin and 296-bp glyceraldehdye 3-phosphate dehydrogenase targets were amplified by reverse transcription-PCR from both sources and the 983-bp glyceraldehdye 3-phosphate dehydrogenase and 499-bp nonhousekeeping integrin-linked kinase targets from frozen tissue. The minimal amount of RNA required for reverse transcription-PCR of 296-bp glyceraldehdye 3-phosphate dehydrogenase target was 29 pg from frozen tissue when RNA subunits were present and 144 pg when these subunits were absent compared to 0.8 ng from paraffin-embedded tissue. Ten laser pulses were required to laser capture sufficient cells from frozen tissue to detect amplification of the 375-bp actin target while more pulses were required for equivalent amplification from paraffin-embedded tissue. Storage time had no detectable effect on RNA quality. We conclude that both frozen and paraffin-embedded tissues as well as laser-captured cells are suitable for gene expression studies but frozen tissue offered greater sensitivity. PMID- 17136988 TI - Initial treatment regimen and risk of hospitalization in patients with chronic obstructive pulmonary disease. AB - STUDY OBJECTIVES: Compare hospitalization risk of various initial treatment regimens for COPD. DESIGN: Retrospective observational cohort design. SETTING: Patients enrolled in 24 different managed care plans across the United States during 1997-2000. PATIENTS: Aged at least 45 years with a primary diagnosis of COPD identified. Initiation date was the date the first inhaler was dispensed. Patients were required to have filled this prescription within 60 days of the first documented COPD diagnosis in the database. INTERVENTIONS: Five therapy cohorts were identified 1) ipratropium alone or in combination with albuterol (IPR), 2) long-acting beta agonists (LABA), 3) inhaled corticosteroid (ICS), 4) ICS+IPR and 5) ICS+LABA. MEASUREMENTS: Subjects were observed for 12 months or until a COPD-related hospitalization was observed, whichever came first. A sensitivity analysis was conducted by varying the observation period from >60, >90 and >180 days to determine if this would impact the results. RESULTS: 3616 patients were identified, 1754 (49%) on IPR alone, 1032 (29%) on ICS alone, 357 (10%) on ICS+IPR, 266 (7%) on LABA alone and 207 (6%) on ICS+LABA. Compared with IPR alone, patients in the ICS alone and ICS plus LABA groups had a 36% and 47% reductions in the risk of a COPD hospitalization, (HR: 0.643; 95% CI 0.512, 0.808 and HR: 0.533; 95% CI 0.328, 0.865) respectively. CONCLUSIONS: The results of this analysis suggest that initial treatment with an ICS alone or in combination with a LABA , compared to IPR alone, was associated with a significant decrease in the risk of COPD hospitalization 12 months following the start of therapy independent of concomitant asthma diagnosis. Similar outcomes were observed when the observation period was varied from >60, >90 and >180 days. PMID- 17136989 TI - Economic impact of pulmonary drugs on direct costs of stable chronic obstructive pulmonary disease. AB - QUESTION OF THE STUDY: To assess the impact of prescribing pulmonary drugs according to the Global Initiative for Chronic Obstructive Lung Disease (GOLD) guidelines on direct costs in stable chronic obstructive pulmonary disease (COPD). PATIENTS AND METHODS: A total of 560 ambulatory COPD patients completed a specific questionnaire that included data regarding drug therapy. Severity was graded according to the British Thoracic Society (BTS) criteria and appropriateness of pharmacological treatment according to GOLD guidelines. RESULTS: Annual direct costs were 1,657 EUR in stage I, 2,425 EUR in stage II, and 3,303 EUR in stage III. The mean direct costs was 2,061 EUR (38% corresponded to drug therapy). Medication accounted for 43%, 37.6%, and 28.4% of total direct costs for stage I, II, and III, respectively. Inhaled steroids and long-acting beta2-agonists accounted for 78%, 76%, and 75% of total drugs costs in stages I, II, and III, respectively. Drug therapy which was not in accordance with guidelines accounted for 78.7% and 54% of total drug costs in stages I and II, respectively. Most patients with severe disease were treated adequately. ANSWER TO THE QUESTION: Pharmacologic treatment has a great impact on direct medical costs in stable COPD. According to GOLD guidelines, patients with mild or moderate COPD are frequently treated with nonrecommended drugs. PMID- 17136990 TI - Factors associated with outcomes of acute exacerbations of chronic obstructive pulmonary disease. AB - The purpose of this article is to provide a general review of the current literature on the factors associated with the outcomes of hospitalizations, survival and health-related quality of life in acute exacerbations of chronic obstructive pulmonary disease (AECOPD), highlighting the limitations and the complexities in interpretation of the results of current studies. There is no consensus definition for AECOPD; onsets may be difficult to define and the determination of duration elusive. The prevalence of acute exacerbations of COPD (AECOPD) in the community appears to be underestimated as exacerbations are underreported by patients and their doctors. Hospitalization for COPD is due mainly to severe AECOPDs which drive the cost of care. There are few longitudinal epidemiological studies on factors associated with hospitalizations for AECOPD. The results of current studies do not allow clear differentiation between associations that are predictors of event, the consequences of the event, or indicators of severity. Strategies to reduce severe exacerbations of COPD include pharmacological treatment, vaccinations, pulmonary rehabilitation, and home care programs. The optimal strategy for the reduction of hospitalization in COPD remains unclear. Long-term interventional studies are needed to provide clearer information for the prevention of exacerbations and hospitalizations in COPD. PMID- 17136991 TI - The workplace impact of acute exacerbations of chronic bronchitis (AECB); A literature review. AB - Acute exacerbations of chronic bronchitis (AECB) are known to have a substantial economic burden in terms of medical care costs. The objective of this study was to assess workplace-based costs associated with AECB, including absenteeism and decreased productivity, based on a review of published literature. A secondary goal was to identify factors related to workplace-based costs in AECB. A literature search was conducted to identify relevant articles assessing one or more aspects of work loss or workplace costs among patients with chronic bronchitis. A review of the identified literature indicates that patients with chronic bronchitis had more days off work; patients whose exacerbations were treated were less likely to have additional exacerbations and had comparatively less work loss. Findings suggest that clinical outcomes and workplace costs are related. While this relationship is clearer in terms of work loss, further exploration is needed to assess decreased productivity and to evaluate this relationship using objective indicators of absenteeism and productivity rather than recall. PMID- 17136992 TI - The role of oxidative stress in chronic obstructive pulmonary disease. AB - Tobacco smoke is the number one risk factor for chronic obstructive pulmonary disease (COPD) and contains a high concentration of oxidants. The lung has a high concentration of antioxidants and antioxidant enzymes; however, COPD patients show evidence of increased oxidative stress suggesting that endogenous antioxidants may be insufficient to prevent oxidative damage from cigarette smoke. The consequences of increased oxidative stress in the lung include increased transcription of inflammatory genes, increased protease activity, and increased mucus secretion. Oxidative stress is often associated with impaired skeletal muscle function and may be one of the causes of glucocorticoid resistance. While current pharmacologic approaches to the treatment of chronic obstructive pulmonary disease do not commonly include antioxidants, preclinical studies involving animal models suggest that antioxidant superoxide dismutase mimetics offer a potential new therapeutic approach to the prevention and treatment of chronic obstructive pulmonary disease. PMID- 17136993 TI - Update on research, diagnosis and management of alphal-antitrypsin deficiency. AB - Formed in response to a World Health Organization recommendation, the Alpha One International Registry (AIR) is a multinational research program focused on alphal-antitrypsin (AAT) deficiency. Each of the nearly 20 participating countries maintains a National Registry of patients with AAT deficiency and contributes to an international database in Malmo, Sweden, that is designed to increase understanding of AAT deficiency as well as safeguard patient confidentiality. AIR members are engaged in active and wide-ranging investigations to improve the diagnosis, monitoring and therapy of the disease. The AIR membership meets biennially to exchange views and research findings. The third biennial meeting was held in Barcelona, Spain, June 11 -13, 2003. A wide range of AAT deficiency-related topics were addressed, encompassing molecular and cellular pathophysiologic mechanisms, clinical epidemiology, diagnostic advances, current and investigational therapeutic approaches, and progress in registry development. Valuable cross-fertilization of concepts and scientific observations was apparent between AAT deficiency research and other fields of biomedicine. The proceedings of the meeting are summarized in this report. PMID- 17136994 TI - National COPD conference summary. AB - The first National COPD Conference, sponsored by the US COPD Coalition was held in Arlington, Virginia on November 14-15, 2003. The theme for the conference was developed around the Department of Health and Human Services (DHHS) Healthy People 2010 goals for COPD and included plenary speeches, roundtable discussions, abstracts, and workshops on spirometry, patient/physician education materials, and home monitoring/telemetry. The goal was to bring together a multidisciplinary group to identify important issues relating to COPD in the United States, specifically the barriers to a wider recognition of the disease, and to develop an orchestrated action plan. Over 500 scientists, clinicians, respiratory therapists, nurses, patients, government officials, and representatives from pharmaceutical companies participated. This summary provides the recommendations from the conference that will be used to develop an action plan for the US COPD Coalition. It includes actions proposed by plenary speakers, roundtable faculty and conference participants. PMID- 17136995 TI - Clinical trial design considerations in assessing long-term functional impacts of tiotropium in COPD: the UPLIFT trial. AB - An accelerated loss of lung function is one of the defining characteristics of chronic obstructive pulmonary disease (COPD). To date, the only successful intervention shown to conclusively attenuate the loss of lung function over time is smoking cessation. Pharmacological interventions including inhaled corticosteroids and ipratropium bromide have not altered the rate of decline of lung function. Tiotropium is an inhaled anticholinergic that provides 24-hour bronchodilation with once-daily dosing due to prolonged muscarinic M3 receptor blockade. Controlled clinical trials have suggested sustained efficacy for periods of up to one year. We therefore initiated a four-year, controlled clinical trial (UPLIFT, Understanding the Potential Long-Term Impacts on Function with Tiotropium) in patients with COPD to evaluate the long-term effects of tiotropium on the rate of decline in lung function and health status as well as the frequency of exacerbations. The design of such large, long-term clinical trials presents unique methodological challenges including the definition of endpoints, the quality and variability of spirometric measurements and premature patient discontinuations from the trial. The present manuscript outlines the rationale for the UPLIFT study, and reviews the study design and the steps taken to address methodological challenges experienced in other long term studies. Careful design and implementation of the UPLIFT trial is anticipated to yield high quality results that will help in increasing our understanding of the long term natural history of COPD in a global population as well as to elucidate the role that tiotropium can play in affecting the course of this debilitating disease. PMID- 17136996 TI - Severe, early-onset emphysema with normal alphal-antitrypsin levels in nonsmokers: a clinical dilemma. AB - Approximately 85% of all cases of chronic obstructive pulmonary disease are attributed to cigarette smoking. The only other established risk factor for the development of chronic obstructive pulmonary disease that is of comparable importance is the deficiency of alpha1-antitrypsin, a rare genetic defect that is present in less than 1% of all cases. Other risk factors are not well characterized in the literature. This article describes one patient without a significant smoking history and with a normal alphaa-antitrypsin level who developed severe early-onset emphysema and gives a brief discussion about other genetic and environmental risk factors for the development of emphysema. PMID- 17136997 TI - The changing role of family physicians in HIV care. PMID- 17136998 TI - Connection between religion and health is complicated issue. PMID- 17136999 TI - Preventing burns in older patients. PMID- 17137000 TI - Acute ankle sprain: an update. AB - Acute ankle injury, a common musculoskeletal injury, can cause ankle sprains. Some evidence suggests that previous injuries or limited joint flexibility may contribute to ankle sprains. The initial assessment of an acute ankle injury should include questions about the timing and mechanism of the injury. The Ottawa Ankle and Foot Rules provide clinical guidelines for excluding a fracture in adults and children and determining if radiography is indicated at the time of injury. Reexamination three to five days after injury, when pain and swelling have improved, may help with the diagnosis. Therapy for ankle sprains focuses on controlling pain and swelling. PRICE (Protection, Rest, Ice, Compression, and Elevation) is a well-established protocol for the treatment of ankle injury. There is some evidence that applying ice and using nonsteroidal antiinflammatory drugs improves healing and speeds recovery. Functional rehabilitation (e.g., motion restoration and strengthening exercises) is preferred over immobilization. Superiority of surgical repair versus functional rehabilitation for severe lateral ligament rupture is controversial. Treatment using semirigid supports is superior to using elastic bandages. Support devices provide some protection against future ankle sprains, particularly in persons with a history of recurrent sprains. Ankle disk or proprioceptive neuromuscular facilitation exercise regimens also may be helpful, although the literature supporting this is limited. PMID- 17137001 TI - Information from your family doctor. Ankle sprains: what you should know. PMID- 17137002 TI - Information from your family doctor. Exercising your ankle after a sprain. PMID- 17137003 TI - Diagnosis and management of granuloma annulare. AB - Granuloma annulare is a benign, asymptomatic, self-limited papular eruption found in patients of all ages. The primary skin lesion usually is grouped papules in an enlarging annular shape, with color ranging from flesh-colored to erythematous. The two most common types of granuloma annulare are localized, which typically is found on the lateral or dorsal surfaces of the hands and feet; and disseminated, which is widespread. Localized disease generally is self-limited and resolves within one to two years, whereas disseminated disease lasts longer. Because localized granuloma annulare is self-limited, no treatment other than reassurance may be necessary. There are no well-designed randomized controlled trials of the treatment of granuloma annulare. Treatment recommendations are based on the pathophysiology of the disease, expert opinion, and case reports only. Liquid nitrogen, injected steroids, or topical steroids under occlusion have been recommended for treatment of localized disease. Disseminated granuloma annulare may be treated with one of several systemic therapies such as dapsone, retinoids, niacinamide, antimalarials, psoralen plus ultraviolet A therapy, fumaric acid esters, tacrolimus, and pimecrolimus. Consultation with a dermatologist is recommended because of the possible toxicities of these agents. PMID- 17137004 TI - Testicular torsion. AB - Each year, testicular torsion affects one in 4,000 males younger than 25 years. Early diagnosis and definitive management are the keys to avoid testicular loss. All prepubertal and young adult males with acute scrotal pain should be considered to have testicular torsion until proven otherwise. The finding of an ipsilateral absent cremasteric reflex is the most accurate sign of testicular torsion. Torsion of the appendix testis is more common in children than testicular torsion and may be diagnosed by the "blue dot sign" (i.e., tender nodule with blue discoloration on the upper pole of the testis). Epididymitis/orchitis is much less common in the prepubertal male, and the diagnosis should be made with caution in this age group. Doppler ultrasonography may be needed for definitive diagnosis; radionuclide scintigraphy is an alternative that may be more accurate but should be ordered only if it can be performed without delay. Diagnosis of testicular torsion is based on the finding of decreased or absent blood flow on the ipsilateral side. Treatment involves rapid restoration of blood flow to the affected testis. The optimal time frame is less than six hours after the onset of symptoms. Manual detorsion by external rotation of the testis can be successful, but restoration of blood flow must be confirmed following the maneuver. Surgical exploration provides definitive treatment for the affected testis by orchiopexy and allows for prophylactic orchiopexy of the contralateral testis. Surgical treatment of torsion of the appendix testis is not mandatory but hastens recovery. PMID- 17137005 TI - Information from your family doctor. Testicular torsion: what you should know. PMID- 17137006 TI - Update on exercise stress testing. AB - Exercise stress testing is an important diagnostic tool for the evaluation of suspected or known cardiac disease. In 2002, the American College of Cardiology (ACC) and the American Heart Association (AHA) revised their guidelines for exercise testing. Ten categories from the ACC/ AHA 1997 guidelines were modified: ST heart rate adjustment, unstable angina, older patients, acute coronary syndromes, chest pain centers, acute myocardial infarction, asymptomatic patients, valvular heart disease, rhythm disturbances, and hypertension. Adjustment of the ST heart rate can identify myocardial ischemia in asymptomatic patients with elevated cardiac risk. Intermediate- and low-risk patients with unstable angina, acute coronary syndromes, or chest pain should undergo exercise stress testing when clinically stable. Provided they are stable, patients who have had acute myocardial infarction can undergo a submaximal exercise test before discharge or a symptom-limited exercise stress test any time after two to three weeks have elapsed. In asymptomatic patients with cardiac risk factors, the exercise stress test may provide valuable prognostic information. Aortic regurgitation is the only valvular heart disorder in which there is significant evidence that exercise stress testing is useful in management decisions. The stress test also can be used in older patients to identify the presence of coronary artery disease. However, because of other comorbidities, a pharmacologic stress test may be necessary. Exercise stress testing can help physicians successfully evaluate arrhythmia in patients with syncope. The exercise stress test also can help identify patients at risk of developing hypertension if they show an abnormal hypertensive response to exercise. PMID- 17137007 TI - Genetic risk assessment and BRCA mutation testing for breast and ovarian cancer susceptibility. PMID- 17137008 TI - Predicting prognosis in patients with end-stage liver disease. PMID- 17137009 TI - A mottled rash on the torso. PMID- 17137010 TI - [What are the most relevant parameters for malnutrition in nursing homes?]. AB - Prevention of malnutrition is an important issue among older nursing homes residents. From this perspective it is crucial that malnutrition is detected in an early stage. Hence, parameters and tools are required that are suitable for these institutions. Based on recent publications an assessment has been made to identify which parameters are most relevant in nursing homes. The literature study indicates that a wide scope of parameters exists for screening of malnutrition. Only a limited number has been studied in the elderly. In addition, more complicated parameters and tools have not revealed substantial added value to relatively simple parameters, such as foodrecords, weight loss and Body Mass Index. PMID- 17137011 TI - [Personal control of care provision and functional performance of non institutionalized and institutionalized elderly]. AB - The present study is part of a broader investigation on the effects of living in a home for the elderly on functional performance and opportunities for successful aging. Functional performance is defined as the daily activities people actually execute in the domains of housekeeping, self-care, and leisure and social activity. In this first study, a comparison was made between community dwelling elderly and aged people living in an institution, in order to investigate the effect of living in an institution on functional performance (FP) in different domains and on personal control (PC) of the received care. The data were gathered in a structured interview, administered to 74 non-institutionalized and 79 institutionalized elderly, aged 71 to 97 years, living in Flanders. We found institutionalized elderly to be less active in every domain of functioning and experiencing less control of the received care. Significant negative correlations between age, FP and PC were only found in the sample of community-dwellers. It seems that in homes for the elderly, care is provided in a package about which the residents have minimal control, which leads to a homogenisation of daily functioning in the institution. PMID- 17137012 TI - [Dutch observation scales to assess cognitive abilities of the aged]. AB - Assessment of complex geriatric health problems by nurses is important for diagnosis, especially assessment of cognitive functioning through daily observations. However, it is unclear which Dutch observation scales are available to assess cognitive abilities. In this study, we present an overview of these scales. A systematic review was performed. Beforehand we determined criteria for inclusion of scales and we searched through Dutch and English databases up till May 2005. Thirteen behavioural observation scales were found. The number of dimensions of cognitive functioning assessed in the scales varied greatly, from two to eight in number. Memory and psychomotor behaviour were always included; consciousness and thinking were frequently included, while alertness, perception, executive functions and language were least included. Extensive assessment of cognitive functioning is highly relevant for a geriatric hospital ward in which patients are admitted for diagnosis. Of all scales that we traced, the A-one is the most extensive: all eight dimensions are included. Little is known about the potential for using the A-one scale in nursing practice; further exploration is indicated. For now, nurses should become acquainted with the different dimensions of cognitive functioning and start to integrate observations in these dimensions in their reporting. PMID- 17137013 TI - [More exercise for seniors: report of program evaluation]. AB - Since 1980, More Exercise for Seniors (MBvO in Dutch) has provided a number of special physical activities for the elderly. The aim is to improve social participation and integration, and to prevent a need for care. Despite the fact that MBvO has been active for more than 25 years, and weekly 300,000 people participate in gymnastics, swimming, dancing, etc, no effect on fitness and health has been proven to date. The study was carried out in two regions: Drenthe and Zuid-Holland. Using the GALM (Groningen Active Living Model) method, 4600 independently living people aged 65 years and older were asked to participate in a motor fitness test. Of the 721 people who participated, 386 (the least fit) were invited to take part in the study. They were than randomly divided into an experimental and a control group. The experimental group did gymnastics once or twice a week. The control group received a health educational program. Assessments were carried out at pretest and posttest (10 weeks). In the groups offered MBvO gymnastics once a week, no effects were found on subjective health, physical performance or quality of life and only minor effects on physical fitness. In the groups offered MBvO gymnastics twice a week, some beneficial quality of life effects were found in the least physically active people at baseline. It is recommended that the frequency and intensity of the regular MBvO gymnastics should be increased, to emphasize the importance of health education, and that relatively inactive and older individuals (70+) should be recruited. PMID- 17137014 TI - [A screening tool to identify older people at risk of adverse health outcomes at the time of hospital admission]. AB - The proportional increase of the ageing population results in an ever growing percentage of elderly among hospitalised patients. Older patients have complex medical, social and psychological problems that could benefit from coordinated care or case management. Identification of high-risk older adults is mandatory to initiate a liaison geriatric management program. A simple screening tool is presented to identify older people at the time of admission who are at increased risk of adverse health outcomes. The instrument was validated during a period of 6 months when all (n = 618) older adults (> 70 year) hospitalised in non geriatric departments of a general hospital were screened. This "Variable Indicative of Placement risk" (VIP) shows a good sensitivity (81%) and specificity (86%) and has a high Negative Predictive Value (97%). Furthermore, it shows a significant positive correlation with the length of stay (p < 0.001). The questionnaire turned out to be a very useful tool in the emergency department as well as in other wards because it probes premorbid frailty components with three simple questions. Due to its simplicity a nurse without geriatric training can complete it. Patients who are not at risk of an adverse outcome are easily recognised. A positive score indicates loss of functional independence and a risk of increased length of stay. Further geriatric assessment and intervention seem then appropriate. PMID- 17137015 TI - Working up policy: the use of specific disease exemplars in formulating general principles governing childhood genetic testing. AB - Non-therapeutic genetic testing in childhood presents a "myriad of ethical questions"; questions which are discussed and resolved in professional policy and position statements. In this paper we consider an underdiscussed but strongly influential feature of policy-making, the role of selective case and exemplar in the production of general recommendations. Our analysis, in the tradition of rhetoric and argumentation, examines the predominate use of three particular disease exemplar (Huntington's disease, Tay-Sachs disease and sickle cell disease) to argue for or against particular genetic tests (predictive testing and testing for carrier status). We discuss the influence these choices have on the type and strength of subsequent recommendations. We argue that there are lessons to be drawn about how genetic diseases are conceptualised and we caution against the geneticisation of medical policy making. PMID- 17137016 TI - Health promotion and the freedom of the individual. AB - This article considers the extent to which health promotion strategies pose a threat to individual freedom. It begins by taking a look at health promotion strategies and at the historical development of health promotion in Britain. A theoretical context is then developed in which Berlin's distinction between negative and positive liberty is used alongside the ideas of John Stuart Mill, Charles Taylor and T.H. Green to discuss the politics of health promotion and to identify the implications of conflicting perspectives on freedom. The final section looks at current health promotion policy in Britain and beyond and argues that, if freedom is seen in terms of empowerment, health promotion can enhance individual freedom. PMID- 17137017 TI - Is health inequality across individuals of moral concern? AB - The history of the documentation of health inequality is long. The way in which health inequality has customarily been documented is by comparing differences in the average health across groups, for example, by sex or gender, income, education, occupation, or geographic region. In the controversial World Health Report 2000, researchers at the World Health Organization criticized this traditional practice and proposed to measure health inequality across individuals irrespective of individuals' group affiliation. They defended its proposal on the moral grounds without clear explanation. In this paper I ask: is health inequality across individuals of moral concern, and, if so, why? Clarification of these questions is crucial for meaningful interpretation of health inequality measured across individuals. Only if there was something morally problematic in health inequality across individuals, its reduction would be good news. Specifically, in this paper I provide three arguments for the moral significance of health inequality across individuals: (a) health is special, (b) health equity plays an important and unique role in the general pursuit of justice, and (c) health inequality is an indicator of general injustice in society. I then discuss three key questions to examine the validity of these arguments: (i) how special is health?, (ii) how good is health as an indicator?, and (iii) what do we mean by injustice? I conclude that health inequality across individuals is of moral interest with the arguments (b) and (c). PMID- 17137018 TI - Consumer choice in Dutch health insurance after reform. AB - This article investigates the scope and effects of enhanced consumer choice in health insurance that is presented as a cornerstone of the new health insurance legislation in the Netherlands that will come into effect in 2006. The choice for choice marks the current libertarian trend in Dutch health care policymaking. One of our conclusions is that the scope of enhanced choice should not be overstated due to many legal and non-legal restrictions to it. The consumer choice advocates have great expectations of the impact of enhanced choice. A critical analysis of its impact demonstrates that these expectations may not become true and that enhanced consumer choice should not be perceived as the 'magic bullet' for many problems in health care. PMID- 17137019 TI - The Quebec complaint examination system: stakeholder perspectives on the purpose and intake of complaints. AB - Quebec's complaint examination system has devoted considerable effort to supporting dissatisfied users who may wish to register complaints. It is open to question, however, whether this level of effort has, in fact, aided users in filing their complaints, and whether, once filed, the intake and processing of complaints has been rigorous and fair. Has the intake and handling of complaints at least improved? This is the question we shall attempt to answer here by presenting the results of our study concerning the impact of the Complaint Assistance and Support Centers (CAAPs) on the intake of complaints. The results show that the Quebec complaint examination system and its Complaint Assistance and Support Centers help make complaints more admissible and ensure that each complaint is examined. However, the system is also hindered by differences and conflicting interpretations among the various stakeholders regarding the legitimacy of complaints, respect for users' rights, and the mission of the system. Although complaint examination systems seek to encourage users to express their points of view, users' voices are still only partly audible. PMID- 17137020 TI - Leib ohne Seele (Body Without Soul). AB - In this brief essay, the Austrian artist Harald Koeck offers an introduction to his painting series "Leib ohne Seele," a selection of which is reproduced. "Leib ohne Seele" is concerned with nature of the autopsy, the people who are the subjects of autopsies, and so with death, and with contemporary attitudes towards and taboos concerning death. While not making an overtly philosophical contribution, Koeck touches on important issues that have been discussed by philosophers as diverse as Martin Heidegger and Peter Singer. Hegel argued that the arts pursue the same goals as philosophy, but do so through sensuous rather than conceptual media. Koeck exemplifies this, allowing his paintings and other works to open up issues for the philosopher. More of his work can be seen at his web site http://www.harald-koeck.com/index.html. PMID- 17137021 TI - The aging males' symptoms in the Italian population: results from a cross sectional study using the AMS scale. AB - OBJECTIVES: To evaluate in the frequency of symptoms associated with the aging male syndrome, using the validated aging male' symptoms (AMS) scale. METHODS AND RESULTS: Men aged 55-85 years were identified by 56 general practitioners (GP) placed in 5 Italian areas among their registered patients during the period May 2005-March 2006, using a computerized generated random numbers list. The GPs were placed in the following areas: Desio, Palmanova/Gorizia in the North of Italy (24 GPs), Parma and Firenze in the Centre (21 GPs) and Napoli in the South of the country (11 GPs). Data were obtained about their general characteristcs and habits. During the interview the men were also asked to fill the AMS scale. Out of the 1927 interviewed men, 1806 men (93.7%) filled correctly the AMS scale. The mean age of the interviewed men was 67 (standard deviation 7.2 range 55-85 years). The symptom most frequently reported as moderate/severe/very severe was: decrease in sexual performance (44.7% of subjects), followed by decreased frequency of morning erection. Among somatic complaints the joint ones were the most frequently reported as moderate/severe/very severe. To be irritable was the most frequent psychological symptoms reported as moderate/severe/very severe. The frequency of all considered symptoms increased with age, but for some somatovegetative and psychological symptoms we observed a "plateau" in the frequency after age 63-70 years. Considering the total AMS score, a severe impairment was reported by 3.5% of interviewed subjects: the values were 1.3%, 3.3% and 6.2% considering separately men aged < or =62, 63-70 and > or =71 years. Considering the frequency of moderate impairment the values were for the total population respectively 17.8% for total score, 21.7% for psychological factors, 30.5% for somatovegetative factors and 32.2% for sexual ones. CONCLUSIONS: This large cross-sectional study gives a general estimate of the frequency of symptoms related with the aging male and clearly indicates that according to the AMS scale a relevant proportion of the interviewed males report moderate severe complaints potentially related with the aging male syndrome. PMID- 17137022 TI - Impact of preoperative biofeedback on incontinence in cancer patients undergoing radical prostatectomy. AB - OBJECTIVE: Post-radical prostatectomy incontinence is mainly attributable to iatrogenic verify the importance of preoperative biofeedback (BFB) in an attempt to identify a strategy to improve postoperative urinary incontinence. MATERIALS AND METHODS: Ninety candidates (median age 68 years: range 49-75) for radical retropubic prostatectomy were evaluated. Forty-five patients underwent preoperative pelvic floor training (PFT) and BFB, whereas the remaining 45 (control group) had preoperative PFT only. Patients started PFT exercises two weeks before surgery and continued during the immediate postoperative period, after catheter removal (6-7 days after surgery), and thereafter at home. No adjuvant therapy was administered and all 90 patients were assessed at follow up visits timed 1, 3 and 6 months post surgery. RESULTS: Patients who were completely dry and without use of pads were defined as continent. At the 1-month follow-up, 6.6% (3/45) of both the PFT+BFB group and the PFT only arm were continent. After 3 months, the continence rate had increased to 33.3% (15/45) and 26.6% (12/45), respectively, and at 6-month follow-up, this percentage had further risen to 71.1% (32/45) in the former group and 66.6% (30/45) in the latter group. No significant differences were observed between the two arms during follow-up, but there was a statistically significant correlation between follow-up time and continence improvement in each group. CONCLUSIONS: Preoperative BFB associated with PFT does not significantly influence urinary continence recovery after radical prostatectomy. PMID- 17137023 TI - p16(INK4a) expression in urinary bladder carcinoma. AB - The most common genetic damage in urothelial carcinoma is partial loss of chromosome 9. The area around 9p21 where the CDKN2A/ARF gene is located is one of the major sites for deletion. This gene encodes for p16 protein which impedes the cell cycle. Specific binding of the p16 product to the cyclin-dependent protein kinases cdk4 or cdk6 inhibits the catalytic activity of the cyclin D-cdk complex, and consequently arrests the cell cycle at the G1/G2 phase. This study aims to immunohistochemically assess p16 expression in urothelial carcinoma in order to evaluate the correlation of this biological marker with tumour stage and/or grade. We studied specimens of transurethral resection (TURB) from 17 cases of non-invasive papillary urothelial carcinoma and from 22 cases of invasive urothelial carcinoma of the bladder. We observed strong p16 immunoreactivity in 11 cases (28.2%), while 28 cases (71.8%) did not show p16 staining. The expression of p16 was statistically associated with disease stage (p = 0.026, according to the chi-square test), but not with either tumour grade or disease progression. These observations call for further studies to focus the importance of p16 expression in bladder cancer development and/or progression. Additional data may provide insight into treatment guided by molecular changes. PMID- 17137024 TI - The role of sexual dysfunctions in inducing the use of drug in young males. AB - INTRODUCTION: Adolescence is one of the periods in which the risk of the initial use of drugs is high. Among the risk factors of first drug use, sexual disorders have been considered one of the possible reasons leading young males to substance abuse and addiction. AIM: To provide data supporting the "La Pera hypothesis", according to which, a very large percentage of young males begin to use drugs due to sexual problems. METHODS: A total of 86 subjects, recruited from 5 drug rehabilitation centers were invited to fill in a questionnaire with their personal data, toxicological history, sexual experiences as well as quality of performances. RESULTS: Approximately 50% of the entire sample used psychotropic drugs to improve their sexual performance with 30 subjects (34.1%) stating that their sexual disorder had influenced their decision to start taking drugs. Of these 30 subjects, 18 (60%) stated their sexual problem had influenced them a little, while in 8 (26.7%), it had influenced them a lot and in 4 (13.3%) it had been a decisive factor. The prevalence of sexual dysfunction prior to first drug use was greater among subjects whose sexual problems influenced their initial use of drugs compared to the remainder of the sample, the difference being statistically significant. CONCLUSIONS: These data are in accordance with the "La Pera hypothesis" based on the cause/effect relationship between sexual dysfunctions and initial use of drugs. It is urgent to demonstrate that sexual disorders lead to first drug use since sex education and early treatment of sexual dysfunctions could provide an effective tool for the primary prevention of substance abuse in young males. PMID- 17137025 TI - Periprostatic tissues intraoperative frozen section during retrograde radical retropubic prostatectomy. AB - OBJECTIVE: To evaluate usefulness of periprostatic tissues intraoperative frozen sections (PTs IFSs) during RRP for prostate cancer, in order to find local extraprostatic neoplastic spreading and to eventually modify intervention and resection limits during surgery. MATERIAL AND METHODS: From January 1998 to June 2004, 259 consecutive patients underwent RRP at our department for clinically organ-confined prostate cancer; PTs IFSs were prospectively performed in all cases at membranous urethra after removal of prostatic apex, at whole neurovascular bundle (NVB) or at fibroadipose tissue subtended between prostatic capsule and NVB during extrafascial or nerve sparing (NS) RRP respectively, at middle portion of Denonvillier's fascia, at detrusor ring after removal of the prostate. IFSs positivity was followed by further excision at the corresponding site during intervention. RRP pathological specimen was handled and examined according to European Association of Urology (EAU) guidelines. Student's t-test and chi-square test were used for statistic analysis, matching patients with or without positive PTs IFSs for bioptic Gleason sum, preoperative serum PSA, clinical stage and lymph nodal involvement. RESULTS: PTs IFSs were positive 75 times in 63 patients out of 259. Pathological stage considering PTs IFSs overlapped 2002 TNM definitive pathological stage in 228 patients. The remaining 31 cases showed PTs neoplastic involvement at IFSs. These latter patients did not show prostatic capsular infiltration at definitive pathology. We demostrated intraoperative extraprostatic cancer spreading that was unrecognizable at definitive pathology. PTs neoplastic spreading changed NS RRP in extrafascial procedure in 17 patients out of 121 with preoperative planned NS RRP There were no significant statistic differences between patients with or without positive periprostatic margins (PMs), regarding preoperative serum PSA, bioptic Gleason sum and clinical stage (Student's t-test); lymph nodal involvement rate was not significantly different in both groups (chi-square test). CONCLUSIONS: PTs IFSs during RRP were feasible and effective in order to achieve better local pathological staging in 12% of patients, to modify planned nerve sparing RRP in extrafascial procedure in 14% of cases and to wide intraoperatively surgical resection margins in 24% of patients. PMID- 17137026 TI - Early discharge of hospitalised patients with community-acquired urosepsis when treated with levofloxacin in sequential therapy. AB - OBJECTIVES: To compare the sequential therapy with levofloxacin with a standard treatment in the management of urosepsis. MATERIALS AND METHODS: It was a multicenter, randomized, open label, pilot trial in community acquired urosepsis carried out in 7 hospitals. Levofloxacin administered 500 mg twice a day intravenously and then orally was compared to piperacillin/tazobactam 4 g/0.5 g three times a day, both in combination with amikacin 7.5 mg/Kg twice a day, for a maximum of 14 days. Amikacin could be suspended between day 3-7. RESULTS: Both treatments were efficacious and well tolerated in the 47 enrolled patients, even tough the time to clinical stability was shorter in the levofloxacin treated patients (3.9 vs. 4.9 days). In the levofloxacin group the sequential therapy was performed in 19 (82.6%) cases, after 4.8 +/- 1.7 days of parenteral administration (100% in the per-protocol population after 5.1 +/- 1.9 days). CONCLUSIONS: The implementation of the sequential therapy in the levofloxacin group allowed an early hospital discharge in patients with urosepsis and a cost saving in the hospital perspective. PMID- 17137027 TI - An unusual presentation of ureteric colic. AB - A patient presented with acute pain in the right iliac fossa and was found to have a small stone in the lower ureter of a crossed ectopic right kidney. This unique presentation highlights the potential pitfalls that may occur when assessing an acute pain in the right iliac fossa and reminds the invaluable role of IVP. PMID- 17137028 TI - The effects of renal ischemia on kidney function in renal cancer conservative surgery. AB - Over the past ten years nephron sparing surgery for renal cancer has been compared to radical nephrectomy for what concern oncological results, even in elective situations but it is necessary to consider functional results (renal function) as well. Clamping renal artery, inducing a temporary renal ischemia that might represent the major known cause of permanent renal damage, is advised for both open and laparoscopic conservative approach. Data from literature show that before the clamping, the patient should be adequately hydrated and receive mannitol. Ischemia should last preferentially less than 30 minutes and kidney cooling should be advised in case of more prolonged ischemia. Artery reclamping should be avoided. Both in warm and cold ischemia, the renal tissue should be perfused by mannitol once the circulation is resumed. PMID- 17137029 TI - Expansion of elderly couples' IADL caregiver networks beyond the marital dyad. AB - Factors influencing expansion of instrumental activities of daily living (IADL) caregiver networks beyond the spouse/partner were studied, using data from the Asset and Health Dynamics among the Oldest Old (AHEAD) nationally representative sample of American elders (ages 70 and older). Analyses were based on 427 Black and White couples in which one partner regularly received IADL assistance; nearly 20% had expanded networks. Logistic regression showed expanded networks were significantly more likely when spouses had IADL or basic personal activity of everyday living (ADL) limitations and help recipients were wives or had numerous IADL or ADL limitations; they also tended to be more common (p <.10) for couples with numerous nearby daughters and help recipients with proxies and those without serious cognitive problems. Network expansion was unrelated to recipients' number of health conditions and Medicaid coverage or couples' ages, marital duration, income, and number of proximate sons. Implications for service programs and caregiving theories of the circumstances linked to IADL assistance from outside the marital dyad are discussed. PMID- 17137030 TI - Generative acts: family and community involvement of older Australians. AB - The article adopts a developmental approach to successful human aging by exploring the concept of generativity in relation to a study of older Australians' lived experiences of involvement in the family and community. Qualitative data, collected through focus group interviews, were analyzed interpretively using recent developments in Erikson's theory of generativity as a framework. As a result, the present study contributes an in-depth understanding of the role of generative acts to the lives of older people. The data provide illustrative support for Erikson's contention of a generativity/stagnation crisis in later life. Involvement in the family and community is seen as a productive and generative activity, which promotes a positive experience of aging. Two further emergent themes are also explored. First, the experiences of study participants illustrate the reciprocal and cyclical nature of grand-generativity, and the importance of intergenerational relationships. Finally, the data contribute to our knowledge of cultural generativity, and in particular the passing on of cultural knowledge through narratives and modeling. PMID- 17137031 TI - The nature of subjective cognitive complaints of older adults. AB - The current study investigated the nature of subjective cognitive complaints of older adults in relation to a broad array of individual cognitive functions known to decline with age. A 60-item questionnaire was developed to examine: (1) whether older adults experience problems with these cognitive functions (problems with cognition); (2) the extent to which these problems have changed over time (changes in cognition); and (3) whether these cognitive decrements interfere with their daily functioning (difficulties with cognition). The cognitive functions examined were attention, processing speed, working memory, executive function, and memory. One hundred eight-two community-dwelling adults (aged 65-92 years) completed the questionnaire. While participants reported problems with, and changes in, each cognitive function, working memory was recorded as most problematic and demonstrated the most change. Yet, although older adults experience problems with cognition and report changes in cognitive functioning over time, these did not necessarily produce difficulties in daily life. This points to the existence of support mechanisms that may assist older adults in cognitively demanding situations. PMID- 17137032 TI - Life review: implementation, theory, research, and therapy. AB - A selective literature review of publications on life review generated ideas on implementation, theory, research, and therapy. The review begins by differentiating life review from reminiscence, and summarizing ways to conduct a life review. A dozen theories that have been influenced by the life review technique are presented, with a focus placed on Erikson's Stages of Psychosocial Development. Erikson's theory has not only been influenced by the life review, it has had a major impact on life review research. Three research topics are discussed: Erikson's ego integrity versus despair in old age, demographical differences in the practice of life reviews, and the impact of conducting life reviews on staff in institutional settings. Most practitioners of life reviews are institutional staff, university students, and family members, many of whom have had limited prior training. The prospect of a certified life review training program is discussed. PMID- 17137033 TI - [The lymph node dissection in thyroid carcinoma]. AB - Rule and indications of lymph node dissection (LD) in thyroid carcinoma is still under debate because of the biological variety of different histological types and the difficulty to have an accurate preoperative diagnosis of metastatic lymph nodes; moreover prognostic factors of metastatic lymph nodes are still unclear. The AA. have separately analyzed different thyroid carcinomas and different clinical situations requiring thyroid LD. Metastatic lymph nodes from differentiated carcinomas, including papillary and follicular type, range from 12 to 90% and apparently do not worsen the prognosis. Level II-VI LD is indicated in presence of metastatic lymph nodes or macroscopic nodal recurrence after a previous LD. No prognostic advantages have been demonstrated when LD is performed without clinical or instrumental evidence of metastatic disease, including suspected recurrence characterised only by plasma Thyreoglobulin increased values. Lymph node metastases from medullary carcinoma range from 25 to 63%. Level II-VI LD is indicated if node metastases are present, whereas prophylactic LD, confined to level VI, is always recommended. Controversies still remain about: 1) LD extension whether it is prophylactic (level VI vs. II- VI) or in case of nodal involvement (levels II- VI monolateral or bilateral), 2) LD indications in case of an increased plasma Calcitonin levels during the follow-up after total thyroidectomy, without clinical or instrumental evidence of nodes involvement. Anaplastic carcinoma represents 5% of all thyroid carcinomas; it is the most aggressive type with an early tendency to invade surrounding organs and to give metastases; prognosis is very poor. LD is indicated only for a palliation in cases with compression syndromes. PMID- 17137034 TI - [Well differentiated thyroid carcinoma: new perspectives and old dilemmas]. AB - The diagnosis of well differentiated carcinoma (i.e papillary carcinoma and follicular carcinoma) represents one of the most challenging issue in thyroid pathology. Aim of the present review is to discuss new perspective and old problems in this topic. Three main subjects are developed, corresponding to: 1) the role of fine needle aspiration versus frozen section examination in pre- or peri- operative diagnosis; 2) the management of small papillary tumour; 3) pathological classification of those tumours indeterminate for papillary or follicular nature. There is general agreement that fine needle aspiration represent the best pre-operative diagnostic tool for thyroid nodules; foremost limits are represented by "not diagnostic" and 'follicular lesion, NOS". The former should be repeated or, if suspicious for papillary lesion, improved with intra-operative apposition cytology; the latter should be deferred to histology with frozen section evaluation reserved to those institution with daily practice on this issue. The management of papillary micro-carcinoma (i.e. papillary carcinoma smaller than 1 cm.) in the setting of an otherwise benign thyroid disease is a matter of debate, since several clinicians suggest to consider these as incidental findings thus avoiding additional treatment. Recently this attitude has been supported by the proposal to regard these lesion as "tumour" and not carcinoma: available data on follow up seems to sustain and favour this approach. There exist a group of well differentiated tumours of the thyroid lacking the criteria to be diagnosed either as papillary (i.e. nuclear grooves, nuclear pseudo-inclusion and nuclear clearing) or follicular (i.e. capsular or vascular invasion) carcinoma; for these lesion, whose behaviour (nodal or blood metastasis) can not be predicted, it has been suggested the term of well differentiated tumour of uncertain malignant potential. Finally it has to be mentioned the possible role of molecular biology in the diagnosis of well differentiated thyroid carcinoma; indeed markers such as RET/PTC or PAX8/PPARgamma, which to date have been employed mainly in basic research, might represent useful diagnostic (and therapeutic) tools in the future. PMID- 17137035 TI - [Useful and limits of the biochemical markers for the diagnosis of thyroid carcinoma]. AB - A series of biochemical parameters are useful for the diagnosis and follow-up of differentiated thyroid carcinomas. The measurement of serum thyroglobulin (Tg) is considered for the post-surgical/radioiodine follow-up of papillary/follicular carcinomas. Other than in basal conditions, the importance of Tg levels during TSH stimulation is underlined, either by discontinuation of L-T4 therapy or by recombinant human TSH test. The finding of undetectable Tg levels during TSH stimulation is highly correlated with clinical remission; otherwise, peak Tg levels > 1-2 ng/ml can be suggestive of recurrence/persistence of the disease. The accuracy of Tg measurements for the detection of metastases seems to be higher when compared to 131-1 whole-body scan. The evaluation of basal serum calcitonin levels is recommended for the screening of medullary thyroid carcinoma (MTC). High basal levels suggest the presence of a tumor but a calcitonin increase can be observed also in parafollicular C cell hyperplasia (CCH) and other extra-thyroidal conditions. The pentagastrin test can by pass this obstacle as the calcitonin response seems to be typical of pathological thyroid C cells. The cut-off value of calcitonin response between patients with MTC and CCH remains to be established; the latter condition, however, being considered by some authors as pre-cancerous. After thyroid surgery the measurement of calcitonin constitutes a valid tool for the detection of cure and/or recurrence of the disease. The screening by means of RET oncogene analysis is also described for patients with MTC with Multiple Endocrine Neoplasia (MEN) type 2 syndrome. PMID- 17137036 TI - [Utility and limits of nuclear medicine in the follow-up of differentiated thyroid carcinoma]. AB - Radionuclide therapy using iodine-131 is widely employed in the work-up of differentiated thyroid carcinoma. After a brief introduction on physical and dosimetry characteristics of radioactive iodine, this paper suggests state of the arts protocol for the ablation of thyroid remnants and for the treatment of metastatic disease. Diagnostic applications of Nuclear Medicine in these tumors are critically evaluated as well, from the use of iodine-131 whole-body scan to 18F-FDG PET and other new diagnostic tools. Finally, a brief description of new radionuclide treatments is reported PMID- 17137037 TI - [Ecographic evaluation of the vitality/fertility in the hepatic hydatidosis as indication for pericistectomy]. AB - Hepatic hydatidosis is still now a frequently observed pathology and the total pericistectomy, for surgical difficulties, often request a subtotal pericistectomy with complications such as biliary fistula, haemorrhage and subphrenic abscesses. The Authors reconsidered their hepatic hydatidosis cases to evaluate the indication to the surgery that in their opinion, should have to consider first of all the functional state of the cysts. Infact, only the vital and the fertil ones, less frequent even if rarely found, should have indications to the surgery, because more likely complicated. The dead and steril ones, instead being asymptomatic, should go under periodic control, since destined to degeneration and calcification. Are also compared the pre-surgery data with the parasitologic exam, to evaluate ETG reliability to determine the functional state of the cyst. Twenty one cysts out of 76 were operated correctly because vital/fertile and in 19 of these the ultrasound indications were correct (90.5%). 55 didn't have indications to the surgery since dead/steril and for 51 (92.7%) the ultrasound indication was correct. So we can say that morphological ultrasound data permitted a correct surgery indication for 70 cyst's on their functional state (93.4% of total). In this way the post-operative complication were reduced of 5%. The Authors found morphologic and/or functional ultrasound error for 6 cysts (7.9%), and in only 3 cases (3.9%) the error were both morphological and functional Infact we believe that a ultrasound morphologic classification should have a functional corrispective for the surgical indication So only the unilocular and multivescicular cysts, vital and fertil one, should have indication to the surgery. On the contrary the solid ones should have an ultrasound follow up and treated by chemotherapy if necessary. PMID- 17137038 TI - [Synchronous and metachronous tumours of colon cancer. A review of 5 years of experience (1999-2004)]. AB - The authors report on 30 synchronous cancer (19%) and 9 metachronous cancer (5.7%) observed in 5 years (1999-2004) in 158 patient operated for colon cancer, defining metachronous cancer a tumor arisen at least 6 months after the first one or further then 5 cm from the anastomosis of the first colon resection. International case records report an incidence of 0.6-14% for synchronous cancer and 1-8% for metachronous cancer. The incidence of synchronous cancer is increasing for the presence of more oncogenic factors in the environment, for the improvement in radiology and endoscopy, for the raise of medium life. In accord with other authors, they show that the main risk factor for the developing of metachronous cancer is the coexistence of colon adenomas at the moment of the diagnosis of the tumor. The various incidence percentages can be explained by different way of diagnosis and classification. PMID- 17137039 TI - [Surgical treatment of fecal incontinence secondary to obstetric trauma]. AB - Fecal incontinence (FI) is associated to elevated costs related to diagnostic work-up, surgical treatment and instrumental follow-up. The real incidence is unknown and prevalence is higher after 45 years with a ratio F:M ratio of 8:1. Frequently FI is due to pelvic damage secondary to obstetric trauma. The Authors analyze surgical treatment results of FI secondary to obstetric trauma evaluating pathogenesis and instrumental diagnostic preoperative work-up. In case of muscular injury, "overlapping" of external sphincter represents the treatment of choice allowing a good medium long term results. In the treatment of patients with more complex injures or after overlapping failures, direct sphincteroplasty are indicated. After multiple surgical failures, or in case of pure neural damage, sacral nerve stimulation, graciloplasty or artificial anal sphincter may be offered by referral centers. PMID- 17137040 TI - [Crohn's disease in the elderly]. AB - AIM OF THE STUDY: To define the characteristics of Crohn's disease in the elderly, basing on our own and other Authors' experience. CASISTIC: During last 10 years, 46 patients affected by Crohn's disease were observed at Emergency Surgery Unit of "Vittorio Emanuele" Hospital in Catania (Italy). Of these patients, 4 (8,6%) were old: 3 with an ileal location and 1 with a colic location of the disease. This latter patient was cured only by medical treatment because of the paucity of clinical manifestations of the disease. The other 3 patients underwent surgical treatment (2 ileocecal resections and 1 ileal resection). RESULTS: Up to date all 4 patients are alive and in good general conditions. Only one patient had recurrence of disease, but recovered after medical treatment. DISCUSSION: Crohns disease in the elderly, although usually similar to the adult form, differs for some aspects. Concerning pathology, in the old patient fibro sclerotic and ulcerative aspects predominate and the colic location is more frequent; clinically, in the elderly the occlusive (two out of our four cases) and haemorrhagic complications are seen more than in young-adults; sometimes the symptoms are poor so that a diagnostic delay occurs, with a possible worsening of prognosis. Regarding the treatment of Crohn's disease it, especially in old patients, must be medical, excluding peculiar situations which need surgical treatment (acute complications, unresponsiveness to drugs). Most frequent operations are segmentary ileal resections and ileocecal resections, as it happened in our experience. Post-operative morbidity and mortality are higher in the elderly, comparing to adult rates, because of more frequent cardiac and respiratory complications, due usually to pre-existing conditions. CONCLUSIONS: Crohn's disease affects old patients in 10-15% of cases. In this age-group it is mandatory to suspect the presence of this disease even if the clinical exam is poor, so that specific diagnostic investigations are carried out in order to avoid lost of time, which can be responsible of an unsuccessful treatment. The treatment is essentially medical; surgery, if necessary, must be as most conservative as possible. PMID- 17137041 TI - [Gastric carcinoid: case report and review of the literature]. AB - Carcinoids are rare tumour of neuroendocrin origin, characterized by argyrophylic silver stain reaction, positive immunohistochemical reaction with neuron-specific markers, a typical growth pattern; they can express different peptides and biogenic amines. Gastric carcinoid tumours have been classified on the basis of susceptibility to gastric trophic stimulus and characteristics mucosal surrounding into 3 different types. This classification is important with respect to therapeutic strategy (type adapted treatment), but the option of endoscopic or surgical treatment is influenced by the impossibility to distinguish clearly the benignity or malignity of these lesions. A review of international literature and a rare case of gastric carcinoid tumour associated with chronic athrophic gastritis, developed in a female 65-years old, with anaemia and treated by subtotal gastrectomy, are presented. PMID- 17137042 TI - A mesenteric window to approach a posterior side leak after laparoscopic assisted stapled strictureplasty for Crohn's disease. AB - Strictureplasty leak in patients with Crohn's disease increases considerably morbidity rate. Intra-operative evaluation of the integrity of suture line and proficient management of any leakage after strictureplasty is mandatory to prevent post-operative complications. When a leakage is detected at the posterior side, through intra-operative insufflation or methylene blue test, it may be difficult to repair as the suture line is completely covered up by mesentery. The AA. describe the creation of a mesenteric window to reinforce the posterior side of a side-to-side stapled strictureplasty performed by laparoscopic assisted procedure. As far as we know, this new technical strategy may allow avoiding resection of small bowel. PMID- 17137043 TI - [Congenital intestinal malformations: clinical evaluation and specific treatment. Case report and literature review]. AB - A case of acute abdomen due to diastasic perforation of the right colon by volvulus on mesenterium commune with associated agenesis of the left kidney and urogenital malformations in a young woman is reported. The case emphasizes the complexity and the polymorphism of congenital abdominal malformations, particularly predisposing to volvulus, that can represent a really challenge for emergency surgeon. The A.A. analyze literature about congenital intestinal anomalies to evaluate clinical, diagnostic and therapeutical problems, and they remark features to take into consideration in emergency: clinical history in non operated patient of abdominal pain and biliary vomit with spontaneous resolutions; nutritional compromise; hypothetical diagnosis of anorexia or food intolerance; malformations in other organs or systems and, one mostly, young age. The correct interpretation of these symptoms can suggest suspicious of an occlusive complication due to intestinal malformations. Radiological data and the exploratory laparotomy or laparoscopy will confirm our intuition. If otherwise diagnosis of the cause underlying the chronic recurrent abdominal pain are delayed, surgical operation in emergency condition will be more aggressive and anatomical structures sacrificed. PMID- 17137044 TI - [Infrequent early complication of mechanical digestive anastomosis: bleeding. Personal experience]. AB - INTRODUCTION: Bleeding from mechanical digestive anastomosis is an uncommon complication (0.9-3.2%) often self-limiting but potentially lethal if not evidenced intraoperatively or in the immediate postoperative. MATERIAL AND METHODS: The Authors retrospectively report incidence of anastomotic bleeding after stapled anastomosis (11/163 = 6.7%) and analyse probable causes. In 6 of 11 patients (54%) intraoperative bleeding was stopped after manual reinforce of anastomosis (3/6) or stopped spontaneously (3/6). In 5 patients (45%), 1 with gastro-jejunal anastomosis, 2 with ileo-colonic anastomosis and 2 with colo rectal anastomosis, they used endoscopy and endoscopic treatment in emergency. RESULTS: All 5 patients were treated with endoscopic clerotherapy (NaCl 0.9% plus epinephrine 1:10000): in 4 (80%) the Authors obtained hemostasis after the first treatment but in one of 2 cases ol ileo-colonic anastomosis (20%) the bleeding relapsed and the patient was re-operated. In 1 patient with the self-limiting lower anastomotic bleeding was associated to a Dieulafoy's gastric ulcer, perendoscopic treated successfully. In summary 2 patients were resubmitted to laparotomy, without evidence of source of bleeding. CONCLUSIONS: In accord with literature, bleeding from mechanical digestive anastomosis is a rare complication, often self-limiting (50-76%), that may be evidenced and treated early in intraoperative phase. Endoscopic examination may have diagnostic (source and type) and therapeutic valence, is effective, with low intrinsic risk and can reach endoscopic hemostasis without relaparotomy, except in case of rebleeding. PMID- 17137045 TI - Inflammatory breast cancer. AB - The very original definition of carcinomatous mastitis suggests the main diagnostic difficulty of this onchologic condition. Actually it is more correctly appointed as inflammatory breast cancer, because notwithstanding the inflammatory appearance, it is a true systemic breast cancer, for which surgery represents an only marginal treatment resource. If is now well known that radical mastectomy is ineffective and only a combination of CT and RT offers survival results of some value. But the actual correct multi modal treatment cannot be undertaken if not after a correct diagnostic confirmation, avoiding an anti-inflammatory therapeutic attempt that is time consuming and possibly deceptive. Surgery and histology can be the only means of a correct diagnosis when FNAB is falsely negative, but the lymph nodes must be the preferential tissue to examinate to avoid long lasting drainage from the breast parenchyma after a biopsy, that delays the beginning of the true treatment. PMID- 17137046 TI - [Progresses and perspectives in cybersurgery]. AB - Aim of this paper is to describe the last progress of surgical robotics owing to the more precise and more reliable instrumentations The surgical robotic applications supported by these technological developments and by the new applications allowed by the outstanding contribution of Electronic Bioengineering Had the possibility to utilise more powerful Telecommunications Networks, essential tool for the data transmission, having an impact in several areas like Telemedicine, Diagnosis and Medical and Surgical Therapy of the patient. The data transmission in real time, that of course is not influenced by the distance, allows a new virtual contact (Map-Volume) and a Clinical three-dimensional Anatomical Space (3D) operative between the surgeon and the complex robotical system. Formed by the Monitor/Controller/Robot Surgeon/patients and distant neighbours. The use of Robotic Surgery, more and more involved in Telemedicine and in the complex system of teleassistance inside the Emergency Centres for all type of catastrophe, will be essential and decisive in the nearer future. In conclusion, many new scenarios with various applications have be opened us for Telerobotic Surgery. Innovations, applications, developments of new systems will involve a greater and greater number of technicians, Doctors, Bioengineers, Clinical Engineers, Informatic Staff in all Telemedicine sectors.. PMID- 17137047 TI - Agricultural researchers and social scientists join medical and veterinary doctors to combat an emerging zoonosis in sub-Saharan Africa. PMID- 17137048 TI - Telemedicine in Africa. PMID- 17137049 TI - Phylogenetic analysis to define feline immunodeficiency virus subtypes in 31 domestic cats in South Africa. AB - Feline immunodeficiency virus (FIV), a lentivirus, is an important pathogen of domestic cats around the world and has many similarities to human immunodeficiency virus (HIV). A characteristic of these lentiviruses is their extensive genetic diversity, which has been an obstacle in the development of successful vaccines. Of the FIV genes, the envelope gene is the most variable and sequence differences in a portion of this gene have been used to define 5 FIV subtypes (A, B, C, D and E). In this study, the proviral DNA sequence of the V3 V5 region of the envelope gene was determined in blood samples from 31 FIV positive cats from 4 different regions of South Africa. Phylogenetic analysis demonstrated the presence of both subtypes A and C, with subtype A predominating. These findings contribute to the understanding of the genetic diversity of FIV. PMID- 17137050 TI - A pilot study on post-evisceration contamination of broiler carcasses and ready to-sell livers and intestines (mala) with Campylobacter jejuni and Campylobacter coli in a high-throughput South African poultry abattoir. AB - To assess post-evisceration contamination of broiler carcasses, 300 samples were randomly selected during routine slaughter in the winter of 2004. The samples originated from 50 chicken carcasses, taken directly after evisceration, as well as 25 samples from ready-to-sell packages of fresh intestines (mala) and livers. The samples were taken in batches over a period of 4 weeks to allow randomised sampling from different farms of origin. Conventional culture-based detection methods of Campylobacter spp. usually need 4-6 days to produce a result. The polymerase chain reaction (PCR) used for this study took less than 32 hours. The average contamination rates with Campylobacter in both the skin and liver samples were 24%, and 28% for intestines. Chicken and chicken products, especially livers and intestines, form an integral part of the traditional diet of many Black South Africans, as they are cheap and readily available in bulk and un-chilled for direct distribution, mainly through street vending and other informal retail outlets. This sudy showed that Campylobacter spp. are prevalent in poultry in South Africa. The handling of poultry meat and products contaminated with this organism in households and the potential for cross-contamination of other foods presents a high risk of infection to consumers in South Africa. The study also emphasised the need for further research in this field. PMID- 17137051 TI - Comparison of morphine and carprofen administered alone or in combination for analgesia in dogs undergoing ovariohysterectomy. AB - In this study the analgesic efficacy of the pure agonistic opioid morphine and the cyclo-oxygenase type-2-selective carprofen were compared since there is no previous specific comparative study for these two common analgesics. Forty-five bitches undergoing elective ovariohysterectomy were randomly assigned to one of three groups; receiving morphine 0.4 mg/kg bodyweight pre-operatively and 0.2 mg/kg every 4-6 hours thereafter (Morphine group), receiving a once-off carprofen 4 mg/kg injection (Carprofen group) or receiving both morphine and carprofen (MorphCarp group). The dogs were premedicated with acepromazine 0.01 mg/kg and induced with either thiopentone 5-10 mg/kg or propofol 4-6 mg/kg. General anaesthesia was maintained with halothane in oxygen. The degree of pain was assessed over a 24-hour period under blinded conditions using a pain scale modified from the University of Melbourne pain scale and the Glasgow composite pain tool. Physiological parameters such as respiratory rate, pulse rate and body temperature were also assessed over the same time period. There was no significant difference in pain-scores and thus analgesia offered by the three analgesia protocols at any assessment point across the three groups, but there were differences within groups across time points. Baseline total pain-scores were lower than scores at all post-operative points within all three groups. Both morphine and carprofen provided good analgesia without any obvious adverse effects. This study indicates that at the dosages indicated above, carprofen administered on its own produces analgesia equal to that produced by morphine and that the two drugs administered together do not produce better analgesia than either drug administered on its own. PMID- 17137052 TI - The use of a probiotic in captive cheetahs (Acinonyx jubatus). AB - Juvenile captive cheetahs (Acinonyx jubatus) often present with diarrhoea that is commonly associated with bacterial infections. A species-specific probiotic containing Lactobacillus Group 2 and Enterococcus faecium was prepared from healthy adult cheetahs. Juvenile cheetahs (n = 27) between 8 and 13 months of age were included in the probiotic trial. The animals were observed prior to and after feeding of the probiotic which was made available for 28 days. Feeding of the probiotic resulted in a significantly increased body weight in the treatment group (P = 0.026), while there was no increase in the control group. A relative improvement in the faecal quality in the probiotic group during the treatment period compared with the pre-treatment (P = 0.0363) and post-treatment (P = 0.004) period was observed. This was accompanied by an absence of blood and mucus in the faeces during the treatment period in the probiotic group. PMID- 17137053 TI - Diseases of indigenous chickens in Bokaa village, Kgatleng district, Botswana. AB - This study examined flock size and management, level of internal and external parasite burden and seroprevalence of antibodies to poultry pathogens in indigenous chickens in Bokaa village, Kgatleng district, Botswana. The mean flock size was 22.6 +/- 6.85 with a range of 11-34. The mean body weights of cocks and hens were 2.28 +/- 0.56 kg and 1.70 +/- 0.38 kg, respectively. Housing and commercial poultry feed were not provided. Ascaridia galli, Heterakis gallinarum and Syngamus trachea were found in some birds. Although the chickens were not vaccinated against any poultry diseases, serum antibodies to Newcastle disease, infectious bursal disease and infectious bronchitis were detected. PMID- 17137054 TI - Investigating the depth of thermal burns in elephants. AB - Histological examination of burn injuries in elephants revealed that the depth was not as severe as expected from clinical observation. Although the actual burn depth was deep, the thickness of elephant skin, especially the dermis, resulted in the lesions being classified as less severe than expected. Examination of skin samples from selected areas showed that most lesions were either superficial (1st degree) or superficial partial-thickness (superficial 2nd degree) burns with the occasional deep partial thickness (deep 2nd degree) wound. These lesions however, resulted in severe complications that eventually led to the death of a number of the elephants. PMID- 17137055 TI - A survey of free-living ixodid ticks on a commercial game farm in the Thabazimbi District, Limpopo Province, South Africa. AB - Free-living stages of ticks on a commercial game farm in the Thabazimbi District, Limpopo Province, South Africa, were collected by drag-sampling with flannel strips during the period September 2003 to August 2004. A total of 5 tick species was collected from 4 sites. Boophilus decoloratus was the most abundant species, followed by Amblyomma hebraeum. Seasonal abundance of the ticks was quantified and an optimum time to implement control measures against the ticks is proposed. PMID- 17137056 TI - Studies on some paraclinical indices on intoxication in horses from freshly cut Jimson weed (Datura stramonium)-contaminated maize intended for ensiling. AB - Monitoring of changes in some blood laboratory parameters in 34 horses after ingesting freshly harvested maize that was to be used for ensiling, heavily contaminated with young Datura stramonium plants, is described. For a 7-day period the following parameters were monitored: haemoglobin content (HGB), red blood cell counts (RBC), white blood cell counts (WBC), haematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), differential white cell counts (DWC), erythrocyte sedimentation rate (ESR), protein fractions, aspartate aminotransferase (AST), lactate dehydrogenase (LDH), total bilirubin (TB), direct bilirubin (DB), blood glucose (Glu), total protein (TP), globulin (Glob) and albumin (Alb). The intoxication was accompanied by erythrocytosis, leukocytosis, regenerative left shift neutrophilia, lymphopaenia, eosinopaenia, increased haematocrit values, low erythrocyte sedimentation rate, hyperglycaemia, bilirubinaemia, hypoproteinaemia and increased activity of AST and LDH. No changes occurred in the mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), albumin, globulin and globulin fractions (alpha 1, alpha 2, beta 1, beta 2 and gamma). The blood parameters returned to normal between post-intoxication days 2 and 5. The observed changes in clinical chemistry indices could be used in the diagnosis, differential diagnosis and prognosis of Jimson weed intoxication. PMID- 17137057 TI - Ipsilateral vascularised ulnar transposition autograft for limb-sparing surgery of the distal radius in 2 dogs with osteosarcoma. AB - Canine osteosarcoma is the most commonly diagnosed primary bone tumour in the dog, affecting mainly large and giant breed dogs with the predilection site being the metaphysis of long bones, specifically the distal radius, proximal humerus, distal femur and proximal tibia and fibula. Treatment options are either palliative or curative intent therapy, the latter limb amputation or limb-sparing surgery together with chemotherapy. This article describes the use of an ipsilateral vascularised ulnar transposition autograft as well as chemotherapy in 2 dogs with osteosarcoma of the distal radius. Both dogs showed minimal complications with the technique and both survived over 381 days following the surgery. Complications seen were loosening of the screws and osteomyelitis. The procedure was well tolerated with excellent limb use. This technique is indicated for use in cases with small tumour size that have not broken through the bone cortex. PMID- 17137058 TI - Magnetic resonance imaging findings of a metastatic chemodectoma in a dog. AB - A 6-year-old, male, Collie-cross was presented with a non-weight bearing right thoracic limb lameness, right m. deltoideus, m. infraspinatus and m. supraspinatus atrophy, and severe neck pain with spasm of the cervical epaxial muscles. MRI revealed complete destruction of the 5th and 6th cervical vertebral bodies with lateral extradural spinal cord compression at the level of the 4th and 5th cervical vertebrae. These lesions were very clearly demonstrated on magnetic resonance images, while only subtle changes were seen on survey radiographs. Post mortem investigation revealed a large heart base chemodectoma with multiple smaller tumours in the cranial mediastinum and a single tumour nodule on the thoracic aorta. The 5th cervical vertebral body had necrotic, haemorrhagic and lytic changes. Histopathology of the heart base tumour, the nodules in the cranial mediastinum and on the thoracic aorta and samples from the 5th cervical vertebra confirmed the presence of a malignant aortic or carotid body tumour originating from the chemoreceptor organs. Diagnostic imaging features and post mortem findings are described. To our knowledge, this is the first report of the magnetic resonance features of a metastatic chemodectoma in a dog. PMID- 17137059 TI - Canine renal cortical necrosis and haemorrhage following ingestion of an Amitraz formulated insecticide dip. AB - Amitraz is a formamidine compound used in veterinary medicine as a topical dip to control ticks and mites on dogs and livestock. A 10-year-old female Scottish terrier was presented following the accidental oral administration of a dip containing amitraz. This case report describes the clinical signs, treatment and pathology of this dog. Clinical signs of toxicity from amitraz result from stimulation of alpha2-adrenergic receptors. Amitraz is seldom fatal because the effects can be reversed by alpha2-adrenergic antagonists. The dog recovered from the amitraz toxicity but died 5 days later from acute renal failure. PMID- 17137061 TI - Problems of evidence-based patient choice in screening programmes. AB - A screening programme is a coordinated set of services for a defined population. Its purpose is to reduce the risk of dying, for example, of breast cancer. But testing does harm as well. For all potential screening programmes policies were developed in the UK during the last ten years: For 22 programmes there are national standards and annual reports, for 17 standards are missing and for 68 cases there is a policy not to screen. Screening is a population based programme, but it is the individual's informed choice whether to take part or not. PMID- 17137062 TI - Communicating "bad news" about screening results to patients. AB - There is much enthusiasm for cancer screening in the United States. In Germany, promotion of cancer screening is growing, often including biased and misleading information. Consequently, the risks of cancer and death from cancer are overestimated, while the harms of screening such as false positive results and over-treatment are underestimated. However, high-quality evidence-based information on breast cancer screening and colorectal cancer screening is available in Germany. PMID- 17137063 TI - The trade-off between population and individual benefit of screening. AB - In cancer screening there is a delicate balance between the benefit for the population and the individual benefit. In different European mammography and other screening programmes major differences were observed regarding detection rates and false positive rates. With massive screening programmes starting in Germany, careful evaluation of their performance and achievement is mandatory, including a determination of the false negative rates. This requires the determination of interval cancer rates on the basis of complete population-based cancer registries. PMID- 17137064 TI - How to assess the methodological quality of systematic reviews of diagnostic trials. AB - It is discussed how the methodological quality of systematic reviews of diagnostic tests can be assessed. Different types of diagnostic questions require different types of systematic reviews. Reviews of diagnostic accuracy are important in choosing the best test for diagnostic confirmation, but information on the diagnostic-therapeutic impact is even more relevant. A thorough assessment of whether a new test is worth doing in the clinical setting requires that primary studies and systematic reviews address the impact on changes in management. PMID- 17137065 TI - New methods in the assessment of diagnostic procedures. AB - In many countries diagnostic tests compose a growing part of healthcare costs, although there are very low standards for their use and for diagnostic studies and evaluations. Reference standards often do not exist. Diagnostic research needs careful consideration of the clinical situation, and should take into account that diagnosis is a multivariable process. Various study designs to evaluate diagnostic procedures are presented. PMID- 17137066 TI - Clinical guidelines: quality and implementation. AB - So far 60 evidence-based clinical guidelines have been developed in Finland. These are no binding rules, but they require interpretation for specific patients, and patient preferences are to be observed. For high-quality guidelines, surrogate markers must be avoided, evidence must be systematically searched for and presented, and local context has to be taken into account. The AGREE (Appraisal of Guideline Research & Evaluation) instrument for evaluation of guidelines is presented. Research on implementation and continuous evaluation of guideline effects are needed. PMID- 17137067 TI - Biased underreporting of research is unethical and should be outlawed. AB - Biased underreporting and multiple or selective publications of clinical trials harm and sometimes kill patients. Publicly funded researchers are not innocent of this form of scientific misconduct, but it is a particular problem in industry sponsored research. So far, only six pharmaceutical companies have endorsed the "Good Publication Practice for Pharmaceutical Companies Guidelines". It is necessary to register all clinical trials publicly, at inception. Drug companies should be forced to publish all results, regardless of outcome. PMID- 17137068 TI - [Is reaching consensus on the development of guidelines nonsense?]. AB - Cultural background influences the development of guidelines. In Germany, classifications exist from S1 (low level of evidence, no structured process of consensus finding) up to S3 (high level of evidence, consensus finding by a representative body). Consensus is indispensable in the guideline development process because every step is influenced by preferences and values, from the choice of output and outcome criteria to the role of pathophysiological models in the assessment of new therapies. A structured process of consensus finding increases transparency. Different models of consensus finding are discussed. PMID- 17137069 TI - [Do we need guidelines to practise better medicine?]. AB - Evidence-based guidelines are recommendations for therapy, but they do not restrict therapeutic freedom. The question is discussed whether more binding directives, including sanctions, are needed to protect patients from superfluous or even harmful treatment. Some key elements of guidelines may be transformed into directives after careful examination of the evidence. A quality assurance system and an evidence-based patient information system are required. PMID- 17137070 TI - [Restrictions to therapeutic freedom or strengthening of medical treatment responsibility]. AB - Freedom of therapy is an important feature of our healthcare system, but at the same time limited by (1) ethical obligations, (2) the interaction between physician and patient, (3) the quality of the professional training, and (4) the availability of resources. Freedom of therapy is less restricted if well trained physicians adhere to high quality guidelines, and hospitals or surgeries are managed efficiently. PMID- 17137071 TI - [How much evidence is needed in the case of fatal diseases and when?]. AB - Spectacular cases of healing of usually fatal diseases gain much attention and put pressure on physicians to apply treatments which have not been sufficiently approved in clinical studies. But often these cases are poorly documented. Also, the number of spontaneous remissions in cancer is greatly overestimated. Promising individual observations must be confirmed by case series in order to collect information which can help future patients. Efforts are necessary to organize well planned studies, especially on rare diseases. PMID- 17137072 TI - [Is there more than one "true" theory of cognition in medicine?]. AB - Alternative approaches such as Traditional Chinese Medicine are very popular with patients. Often, however, there is a lack of high quality studies to prove their efficacy. The question will be discussed whether different theories of cognition may coexist in medicine which are all true. Not only biological, but also psychological and social factors contribute to health and well-being. Integrative medicine should bridge the gap between mainstream evidence-based medicine and complementary methods. Shared decision making between patient and physician can help to find the optimum, often multimodal treatment strategy for each patient. PMID- 17137074 TI - [In]. PMID- 17137073 TI - [Can individual experience be considered for efficacy assessment?]. AB - The question is discussed whether the efficacy assessment of a therapy can be modified by the observation of a different outcome in individual cases. N-of-1 studies may help to decide between different options in special individual circumstances. Case series have their role in medicine, especially for generating hypothesis. But the observation of isolated cases can be no substitute for efficacy assessment through clinical trials, which are based on a systematic compilation and statistical analysis of many individual cases. PMID- 17137075 TI - [The neuroprotective B2 strengthens the aggregation of unfolded protein in Huntington chorea and Parkinson disease]. PMID- 17137076 TI - [A genetic marker for type 2 diabetes?]. PMID- 17137077 TI - [A definition of overweight and obesity]. PMID- 17137078 TI - [The epidemiology of obesity]. PMID- 17137079 TI - [Obesity in children and young adults]. PMID- 17137080 TI - [Pharmacotherapy of obesity]. PMID- 17137081 TI - [Interdisciplinary obesity treatment]. PMID- 17137082 TI - [Selective cannabinoid receptor antagonists]. PMID- 17137083 TI - [Nutritional supplements in overweight and obesity]. PMID- 17137084 TI - [Obesity form the health care economics point of view]. PMID- 17137085 TI - [Congress: chemistry, pharmacology and biosynthesis of alkaloids]. PMID- 17137086 TI - [New findings on the chemistry, pharmacology and clinical uses of purines]. PMID- 17137087 TI - [Pharmaceutical science in Europe]. PMID- 17137088 TI - [New trends in HPLC in 2006]. PMID- 17137089 TI - [Sore throat and a swollen neck: Lemierre's syndrome until proven otherwise]. AB - Physicians seldom prescribe antibiotics to patients presenting with a combination of sore throat and feelings of malaise. However, this restrictive regimen may have a downside. Two patients, men aged 23 and 19 years, respectively, with pharyngitis developed a life-threatening syndrome following a Fusobacterium throat infection. They suffered from Lemierre's syndrome, referred to in recent literature as the 'forgotten disease'. The second patient recovered uneventfully after prompt intravenous antibiotic treatment. The first, however, succumbed following overwhelming multiple organ failure. A classical case of Lemierre's syndrome is characterised by bacterial embolisation from a clogged internal jugular vein following a sore throat. Spread of organisms may lead to sepsis and organ failure. The main pathogen is Fusobacterium necrophorum. Early recognition is essential, since prompt antibiotic treatment is usually effective. This syndrome should be suspected until proven otherwise in any patient with signs of pharyngitis, a painful swollen neck and pulmonary symptoms. PMID- 17137090 TI - [The cost per year of life gained: trends and internal contradictions]. AB - Recently, a Dutch advisory committee to the Ministry of Health presented a report proposing criteria for health insurance coverage. A central role has been given to the criterion of cost per quality-adjusted life year (QALY) associated with medical interventions. The proposal is to limit the health-insurance coverage of such costs to 80,000 euro per QALY gained. Previously, 20,000 euro was generally considered a reasonable critical level for QALY-cost, whether for preventive or for curative interventions. Although a higher cut-off-level is now proposed by the advisory committee, recent cardiovascular prevention guidelines propose treatment criteria that are associated with much lower QALY costs, sometimes as low as 1,000 euro. With respect to financial considerations, the central role of QALY costs has been replaced in these prevention guidelines by national health budget implications. For individual patient care, these apparently opposing trends in the role and financial appreciation of QALYs have complex repercussions. PMID- 17137091 TI - [Cancer registries and the quality of oncological care]. AB - In 1971, President Nixon of the US declared war on cancer. To ascertain what progress has been made since then, national cancer registries may provide informative data. However, if standardised morbidity and mortality summary rates for cancer are dropping, we cannot conclude that there is a causal relation between increased diagnostic or therapeutic efforts and this decline. Cancer is a biologically complex disease with more than 400 different histological types of tumour. Many different efforts are made simultaneously to improve diagnostic accuracy and therapeutic effectiveness in selected types of malignancies. Since most cancers occur in elderly people, comorbidity is also a frequent problem. Final figures are therefore influenced by many different factors. Cancer registries reflect the result of all these factors, but figures indicating progress in the war on cancer have to be interpreted with caution. PMID- 17137092 TI - [Deep-vein thrombosis and pulmonary embolism due to air travel]. AB - In recent years, the association between air travel and the incidence of deep vein thrombosis or pulmonary embolism has become clearer. Epidemiologic studies reveal an increased relative risk of thromboembolism after flights of more than 8 hours and especially in subjects at higher risk for this disease, due, for example, to congenital thrombophilia or the use of oral contraceptives. However, the absolute risk of deep-vein thrombosis or pulmonary embolism after prolonged air travel is very small. Studies have shown that a combination of factors present during prolonged air travel may account for increased activation of coagulation. There is no definitive proof that elastic stockings are effective in reducing the incidence of clinically relevant thromboembolism during air travel. Acetylsalicylic acid is not effective in the prevention of thrombosis during air travel and may be dangerous. Prophylactic subcutaneous low molecular weight heparin may be effective to prevent air travel-associated thrombosis. However, pending more solid evidence, this strategy should only be used cautiously, e.g. only in patients with a high risk of thrombosis who are planning a long flight. PMID- 17137093 TI - [Subfertility in overweight women]. AB - The prevalence of overweight individuals in The Netherlands is increasing sharply as has also been observed in populations worldwide. In addition to the long-term health risks of being overweight, overweight women of reproductive age are more commonly faced with reproductive disorders. Women who are overweight are less fertile than women of normal weight. The chances of both spontaneous conception and conception after ovulation induction and assisted reproduction are lower in women who are overweight. The chance of a live birth is also decreased due to an increased risk of miscarriage. Furthermore pregnancy outcome is compromised by obesity-related complications of pregnancy. Weight loss of 5-15% in subfertile women who are overweight increases the chance of spontaneous conception and conception after fertility treatment and can be achieved through a low-calorie diet, increased exercise and behaviour modification. PMID- 17137094 TI - [Levels of evidence value are useful in the evaluation of studies and results]. AB - A classification of scientific evidence into 'levels', in which the type of research design is graded according to its strength, is useful because it helps the medical community to sort out a system that is accepted all over the world. It not only makes the highest levels of evidential value explicit for a particular clinical question, but also makes the lack of evidence visible. Selected articles will contain labels identifying the strength of evidence supporting key recommendations. PMID- 17137095 TI - [Levels of evidence are insufficient]. AB - Medical knowledge is based on various types of research, each with its own 'indication' and 'contraindication'. Although the randomised controlled trial is highly useful to quantify small differences in treatment effects, it is not able to establish all medical knowledge needed at the bedside. In most instances of non-therapeutic research, observational data are more useful. Evaluating all medical knowledge on the basis of a single hierarchy of level of evidence is therefore seriously flawed. PMID- 17137096 TI - [From the Cochrane Library: wearing compression stockings during long-distance flights reduces the risk of asymptomatic deep-vein thrombosis]. AB - A recent Cochrane systematic review assessed the effects of wearing compression stockings during long-distance flights on the risk of deep-vein thrombosis (DVT). Ten randomised trials were included, 9 of which compared wearing compression stockings on both legs during the whole flight (at least 7 hours) with not wearing compression stockings. In these 9 trials, 50 of 2637 participants with 1 week of follow-up had asymptomatic DVT; 3 had worn stockings, 47 had not (odds ratio: 0.10; 95% CI: 0.04-0.25). No deaths, pulmonary emboli or symptomatic DVTs were reported. Wearing stockings had a significant impact in reducing oedema of the lower legs. No significant adverse effects of wearing stockings were reported. PMID- 17137097 TI - [Diagnostic image (299). An oncology patient suffering from conjunctivitis]. AB - A 64-year-old man with a small-cell bronchus carcinoma and metastases was treated with chemotherapy and developed conjunctivitis and necrosis of the eyelids caused by Pseudomonas aeruginosa. PMID- 17137098 TI - [Trends in the incidence and death from cancer from 1989-2003 in The Netherlands]. AB - OBJECTIVE: To provide insight into the changing nature and size of the cancer burden within The Netherlands. DESIGN: Retrospective. METHOD: Data on incidence and death relating to various forms of cancer are calculated on the basis of registered data concerning the incidence (Netherlands Cancer Registration; NCR) of and death (Statistics Netherlands) from cancer in the Netherlands from 1989 until 2003. RESULTS: From the start in 1989 up to 2003, more than one million new cases of cancer were registered with the NCR. The total number of new patients with a primary tumour increased from 56,335 in 1989 to 73,188 in 2003 (30%). The most frequently occurring tumours in 2003 were of the breast, colon, lung and prostate. The age standardized incidence rate for males and females combined, increased from 381 per 100,000 in 1989 to 400 per 100,000 person years in 2003 (+5%). There was an increase in breast, prostate, skin and oesophagus cancer, and also lung cancer in females. Major decreases were seen in lung cancer in males, as well as stomach, ovary and gallbladder cancer. The number of cancer deaths in the Netherlands increased from 35,420 in 1989 to 38,454 in 2003 (+8%). The age and sex standardized mortality rate declined from 234 per 100,000 in 1989 to 201 per 100,000 in 2003 (-14%). CONCLUSION: Despite a slight increase in the incidence of cancer and an increase in mortality from lung cancer (in females), oesophageal cancer and melanomas, the death rate from cancer has dropped considerably. The changes in incidence and mortality may be explained by changes in lifestyle in the 1970s and 80s, in particular use of tobacco and alcohol. Also early detection and screening programmes have resulted in an increase in the incidence of tumours with a better prognosis, which has led to a decrease in mortality. The downward trend in mortality was also influenced by treatment improving prognoses. PMID- 17137100 TI - [A girl with hereditary myotonia due to an exceptional sodium channel mutation]. AB - A 22-month-old girl had cramps and stiffness of her muscles. After medical history, physical examination and an EMG, a short differential diagnosis based on the symptoms of myotonia was made. Initially, the symptoms were incorrectly assumed to be due to Becker's myotonia, an autosomal recessive condition caused by a mutation in the chloride channel. Molecular analysis did not show a defect in the chloride channel, but instead a defect in the sodium channel of the muscle fibre. Since defects in the sodium channel are responsible for several myotonic diseases, further analysis was necessary. Based on knowledge of the structure and mechanism of the sodium channel and study of literature on cases involving the identical mutation, the diagnosis 'potassium-aggravated myotonia' (PAM) was made. Re-evaluation of the patient showed that her symptoms fitted the diagnosis 'myotonia permanens', the severest form of PAM. She was treated with mexiletine. In myotonia several features can give direction to the diagnosis, including sensitivity to temperature and exercise, and family history. However, it is often necessary to use molecular analysis to be able to diagnose the disease correctly, make a prognosis and predict the risk of recurrence as well as to formulate a treatment plan. PMID- 17137101 TI - [The current MRSA policy in the Netherlands]. PMID- 17137102 TI - [Initiatives to increase the efficiency of dermatological patient care]. PMID- 17137103 TI - Variation of taxane content in needles of Taxus x media cultivars with different growth characteristics. AB - Needles from 17 different Taxus x media cultivars, belonging to 4 groups showing different growth characteristics, were analyzed using high performance liquid chromatography for their content of 10-deacetylbaccatin III, baccatin III, cephalomannine and paclitaxel (Taxol). The 4 Taxus x media cultivar groups were: 1.) medium to fast growing and upright form; 2.) slow growing and upright form; 3.) fast growing and spreading form; and 4.) slow growing and spreading form. The purpose of this study was to identify yew cultivars of fast growth rate, upright growth and high taxane content in their needles. The highest content of paclitaxel was found in 'Coleana' of group 1 (378 microg/g of the extracted dry weight). Three cultivars in group 1, 'Coleana', 'Stovekenii' and 'Hicksii', make good candidates for taxane extraction because of their high paclitaxel and 10 deacetylbaccatin III content, fast biomass accumulation and upright growing form. They are also good starting materials to develop alternative methods for the production of paclitaxel and its analogous compounds through modern biotechnology approaches. PMID- 17137104 TI - Biological evaluation of curcumin and related diarylheptanoids. AB - Nine derivatives of three natural diarylheptanoids, curcumin, demethoxycurcumin and bisdemethoxycurcumin, were prepared. Their antioxidant, free radical scavenging, nitric oxide (NO) inhibitory and cytotoxic activities were evaluated and compared with those of the respective natural compounds. Curcumin (1), demethoxycurcumin (2), demethyldemethoxy-curcumin (C3), diacetyldemethoxycurcumin (AC2) and triacetyldemethylcurcumin (AC5) exhibited higher antioxidant activity than quercetin while products from demethylation of 1 and 2 exhibited higher free radical scavenging activity. Compounds AC2 and AC5 were found to be most active in inhibiting breast cancer cells (MCF-7) proliferation with IC50 values of 6.7 and 3.6 microM, respectively. The activity of AC2 is almost doubled and of AC5 almost tripled as compared to curcumin. Their selectivity towards different cell lines is also more noticeable. Compounds AC2 and AC5 also showed increased activity against a human prostate cancer cell line (DU-145) and non-small lung cancer cell line (NCI-H460) with IC50 values of 20.4, 16.3 and 18.3, 10.7 microM, respectively. PMID- 17137105 TI - Antiviral and antimicrobial assessment of some selected flavonoids. AB - In the current study, the results of antibacterial, antifungal, and antiviral activity tests of four flavonoid derivatives, scandenone (1), tiliroside (2), quercetin-3,7-O-alpha-L-dirhamnoside (3), and kaempferol-3,7-O-alpha-L dirhamnoside (4), are presented. Antibacterial and antifungal activities of these compounds were tested against Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Acinetobacter baumannii, Staphylococcus aureus, Bacillus subtilis, and Enterococcus faecalis, as well as the fungus Candida albicans by a micro-dilution method. On the other hand, both DNA virus Herpes simplex (HSV) and RNA virus Parainfluenza-3 (PI-3) were employed for antiviral assessment of the compounds using Madin-Darby bovine kidney and Vero cell lines. According to our data, all of the compounds tested were found to be quite active against S. aureus and E. faecalis with MIC values of 0.5 microg/ml, followed by E. coli (2 microg/ml), K. pneumoniae (4 microg/ml), A. baumannii (8 micro/g/ml), and B. subtilis (8 microg/ml), while they inhibited C. albicans at 1 microg/ml as potent as ketoconazole. However, only compound 3 displayed an antiviral effect towards PI-3 in the range of 8-32 microg/ml of inhibitory concentration for cytopathogenic effect (CPE). PMID- 17137106 TI - Antimicrobial activity of catechol and pyrogallol as allelochemicals. AB - Catechol and pyrogallol are allelochemicals which belong to phenolic compounds synthesized in plants. Their antimicrobial activities were investigated on three bacteria (Pseudomonas putida, Pseudomonas pyocyanea, Corynebacterium xerosis) and two fungi (Fusarium oxysporum, Penicillium italicum) phytopathogenic species as test organisms using the disc diffusion method. Both catechol and pyrogallol were found to have antibacterial effects on all the bacteria used in the study at 5 and 10 mM concentrations. Catechol has also been found to have an antifungal effect on the fungi used in the study, whereas no antifungal effects of pyrogallol were observed. The most sensitive species among the bacteria was P. putida which was inhibited by the allelochemicals even at 1 mM concentration. PMID- 17137107 TI - Studies on the cytotoxicity of cucurbitacins isolated from Cayaponia racemosa (Cucurbitaceae). AB - The cytotoxic potential of three cucurbitacins, 2,3,16,20(R),25-pentahydroxy 11,22-dioxo-cucurbita-5-en (cucurbitacin P, 1), 2,3,16,20(R),25-pentahydroxy-22 oxocucurbita-5-en (2) and 2,3,16,20(R),25-pentahydroxy-22-oxocucurbita-5,23(E) diene (deacetylpicracin, 3), obtained from Cayaponia racemosa was evaluated as their ability to induce brine shrimp lethality, to inhibit the development of sea urchin eggs and tumor cell proliferation, and to lysis mouse erythrocytes. Compounds 1 and 2 were highly toxic with LC50 of (29.6+/-.1) (56.8) and (38.8 +/ .0) (76.6) micro/mL (umicro), respectively, while compound 3 was not effective at the tested concentrations. All tested compounds possessed an inhibitory effect on the proliferation of tumor cell lines, compound 1 being the most active, followed by 2 and 3. Nevertheless, no hemolytic activity or inhibition of the development of sea urchin eggs was observed for these compounds. PMID- 17137108 TI - Sesquiterpene lactones from Dimerostemma species (Asteraceae) and in vitro potential anti-inflammatory activities. AB - Two Brazilian species of Dimerostemma (Asteraceae) were chemically investigated. Two known sesquiterpene lactones (STLs), a germacrolide and an eudesmanolide, were isolated from D. episcopale while D. brasilianum afforded the new germacranolide 1beta,5beta/10alpha-trihy-droxy-8alpha-angeloyloxy-germacra 3,11(13)-dien-6alpha,12-olide in addition to a known one. Structure identification based on NMR and MS analyses. 1beta,10alpha,4alpha,5beta-Diepoxy 8alpha-angeloyloxy-costunolide isolated from D. brasilianum was studied for its anti-inflammatory activity. This STL completely inhibited DNA binding of the transcription factor NF-kappaB at a concentration of 5 microm and 10 microM in Jurkat T and Raw 264.7 cells, respectively. Elastase release from human neutrophils was reduced to 50% at a concentrations of 23 microM after stimulation with PAF and of 27 microM after stimulation with fMLP without showing cytotoxic effects. Additionally, elastase was also directly inhibited. PMID- 17137109 TI - Cytotoxic activity of orsellinates. AB - The series of 2,4-dihydroxy-6-methylbenzoates 2-10 (methyl to hexyl orsellinates) prepared by alcoholysis of lecanoric acid (1)--a natural product from the lichen Parmotrema tinctorum (Nyl.) Hale - was submitted to the brine shrimp lethality test (BST), which was also performed for 2,4-dihydroxy-6-methylbenzoic acid (11) (orsellinic acid) and the derivative ethyl-2-hydroxy-4-methoxy-6-methylbenzoate (12) (4-methoxy-ethyl orsellinate), in order to detect new substances with probable antineoplasic activity. Results showed that chain elongation--increase in lipophilicity (log P)--causes a rise in the cytotoxic activity of orsellinates. Hexyl orsellinate (7) showed the highest cytotoxic activity (LC50 = 31 microM). A correlation between lipophilicity (log P) and cytotoxic activity (log 1/LC50) is presented. Compounds with ramified chains--iso-propyl, sec-butyl and tert-butyl orsellinates (8-10)--were less active than those with the correspondent linear chain. The activities presented by 4-methoxy-ethyl orsellinate (12) and ethyl orsellinate (3) suggest that the hydroxy group at the C-4 position causes effect in the cytotoxic activity of orsellinates against Artemia salina. PMID- 17137110 TI - Deoxypodophyllotoxin content and antioxidant activity of aerial parts of Anthriscus sylvestris Hoffm. AB - Deoxypodophyllotoxin content of the aerial parts of Anthriscus sylvestris Hoffm. growing at different altitudes was evaluated in comparison to the roots. The lignan accumulation in ground parts was at least double compared to aerial ones. In addition antioxidant-guided fractionation of the crude methanol extract of aerial parts was performed with the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test. Active fractions contained mainly luteolin-7-O-glucoside and chlorogenic acid. Antioxidant properties of both crude extract and isolated compounds were also investigated with the Briggs-Rauscher (BR) oscillating reaction. A satisfactory agreement between the results obtained with the two methods was observed. PMID- 17137111 TI - New bioactive sesquiterpenes from Ripartites metrodii and R. tricholoma. AB - The metabolites of two different Ripartites species, R. tricholoma (A. & S. ex Fr.) Karst. and R. metrodii Huijsm. were investigated. Three new sesquiterpenes were isolated from three different strains. In addition, the strains produced 13 oxo-9(Z),11(E)-octadecadienoic acid, psathyrellon A, 5-desoxyilludosin, an illudane (previously isolated from a Bovista sp.) 96042 and demethylovalicin, five known compounds. PMID- 17137112 TI - Hydroxamic acids in Secale cereale L. and the relationship with their antifeedant and allelopathic properties. AB - Contents of the hydroxamic acids 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA), and 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) in leaves and roots of 14 cultivars of rye, Secale cereale L., were determined. Dynamics of accumulation in three cultivars were evaluated. DIBOA was the main cyclic hydroxamic acid in leaves but the contents differed significantly between the cultivars. Both DIBOA and DIMBOA were present in the roots. Maximum concentration of DIBOA in leaves and DIMBOA in roots was reached between 48-54 h and 54-72 h after germination, respectively. Antifeedant activity of DIBOA towards the aphid Rhopalosiphum padi and the feeding behavior were studied by electronic recording in barley leaves treated with different contents of DIBOA. The deleterious activity of DIBOA could arise by starvation and/or a toxic effect. Additionally, allelopathic potential of pure DIBOA and aqueous extracts of leaves and roots of rye (Tetra-Baer) on the germination of lettuce (Lactuca sativa) and rye (Tetra-Baer) seeds was evaluated. A high percentage of germination inhibition of pure DIBOA and the extracts of leaves and roots was observed. The activity is in agreement with the contents of hydroxamic acids in the plants. The substrates had no allelopathic effect on rye seeds. PMID- 17137113 TI - Antibacterial activity and composition of the essential oil of Ziziphora clinopodioides subsp. bungeana (Juz.) Rech. f. from Iran. AB - The chemical composition of the essential oil obtained from the aerial flowering parts of Ziziphora clinopodioides subsp. bungeana (Juz.) Rech. f. was analyzed by GC and GC-MS. Thirty-two components representing 97.1% of the total oil were identified. Oxygenated monoterpenes (94.3%) were the predominant fraction of the oil with pulegone (65.2%), isomenthone (11.9%), 1,8-cineole (7.8%) and piperitenone (6.5%) as the main constituents. Antibacterial activity of the oil and also its two main components (pulegone and 1,8-cineole) were tested against seven bacteria. It was found that the oil exhibited interesting antibacterial activity against Staphylococcus epidermidis, S. aureus, Escherichia coli and Bacillus subtilis with MIC values of 3.75 mg/ml. PMID- 17137114 TI - Antimicrobial activity and composition of the essential oil of Gontscharovia popovii from Iran. AB - The aerial parts of Gontscharovia popovii (B. Fedtsch. and Gontsch.) Boriss. were collected at full flowering stage. The essential oil was isolated by hydrodistillation and analyzed by a combination of capillary GC and GC-MS. Thirty one components were identified with the main constituent being carvacrol (71.9%), followed by linalool (5.5%), p-cymene (4.5%) and gamma-terpinene (4.4%). The in vitro antimicrobial activity of the essential oil of G. popovii was studied against seven Gram-positive and Gram-negative bacteria (Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae) and three fungi (Candida albicans, Saccharomyces cerevisiae and Aspergillus niger). The results of the bioassays showed that the oil exhibited strong antimicrobial activity against all the tested fungi and bacteria except for the resistant bacterium Pseudomonas aeruginosa. PMID- 17137115 TI - Phytochemical and biological investigation of Aristolochia maurorum L. AB - Aristolochia maurorum L. of Jordanian origin has been investigated phytochemically, quantitatively, and biologically. Three atypical alkaloids, namely aristolochic acid I (1), aristolochic acid II (2) and aristolochic acid IIIa (3), have been isolated and identified. Of these known 1 phenanthrenecarboxylic acids, 2 and 3 are reported for the first time from this species. The identified compounds 1-3 were first evaluated biologically as cytotoxic agents against the brine shrimp lethality test (BST), in which compound 1 was found to be the most potent (LC50, 4.9 microg/mL). The antiplatelet activity of the methanolic extracts, the acidic fractions of aerial and root parts, and the identified compounds 1-3 were evaluated using an automatic platelet aggregometer and coagulation tracer (APACT 2). Using external reference standards, and a reverse-phase isocratic method, the distribution of aristolochic acid I and aristolochic acid II in different plant parts of Aristolochia maurorum L. during flowering stage was analyzed by PDA-HPLC. A quantitative comparison between two previously reported extraction methods was also made. Roots were found to be the main storage of aristolochic acid I and aristolochic acid II during flowering stage with about 0.22 and 0.108% (w/w), respectively. PMID- 17137116 TI - RT-PCR analysis and stress response capacity of transgenic gshI-poplar clones (Populus x canescens) in response to paraquat exposure. AB - Stress response capacity (Fv/Fm at 690 nm and F690/F735 at Fmax) of untransformed hybrid poplar, Populus x canescens (P tremula x P alba), and two transgenic lines overexpressing gamma-ECS (gamma-glutamylcysteine synthetase) either in the cytosol (cyt-ECS) or in the chloroplast (chl-ECS) was studied in response to the herbicide paraquat (4.0 x 10(-9) to 4.0 x 10(-6) M) for 21 days. Significant differences at sublethal (4.0 x 10(-7) M) and bleaching (4.0 x 10(-6) M) concentrations of paraquat were observed with about a two-fold and eight-fold decrease in the photosynthetic activity (Fv/Fm at 690 nm and F690/F735 at Fmax), respectively. None of the gshI transgenic lines (cyt-ECS, chl-ECS) with elevated GSH content exhibited significant tolerance to paraquat. Semiquantitative RT-PCR of the cyt-ECS clone was used for gene expression analysis of the nuclear encoded rbcS gene and the stress responsive gst gene. Expression of the constitutively expressed 26SrRNA ribosomal gene was probed as a control for all RT-PCR reactions. The relative intensities of gene expressions normalized to the level of 26SrRNA intensity showed a 50% decrease in the nuclear encoded rbcS expression and a 120% increase in the stress responsive gst gene expression of the paraquat treated (4.0 x 10(-7) M) samples of the transgenic poplar line (cyt-ECS). PMID- 17137117 TI - Relationship between grain yield, osmotic adjustment and benzoxazinone content in Triticum aestivum L. cultivars. AB - Fifteen wheat genotypes were grown under water deficit to ascertain the role of osmotic adjustment (OA) and the concentration of benzoxazinones in sustaining grain yield. A positive correlation between osmotic adjustment capacity and yield was observed in wheat genotypes cultivated under field conditions. The weight gain of plants exposed to drought was in agreement with the OA values (R(2) = 0.93). However, when wheat plants were infested by cereal aphids, this correlation was not found. The benzoxazinones 2,4-dihydroxy-1,4-benzoxa-zin-3-one (DIBOA) and 2,4-dihydroxy-7-methoxy-1,4 benzoxazin-3-one (DIMBOA) are defensive secondary metabolites present in wheat and others cereals. The content of these compounds varied in wheat genotypes and increased with drought and aphid infestation. A positive correlation between weight gain of irrigated-infested plants and drought-infested plants and the contents of benzoxazinones was observed. These results suggest that plants with better OA capacity and high benzoxazinone content should have better field yields. PMID- 17137118 TI - Effect of 2-benzoxazolinone (BOA) on seedling growth and associated biochemical changes in mung bean (Phaseolus aureus). AB - BOA (2-benzoxazolinone) is a potent phytotoxin present in several graminaceous crops such as rye, maize and wheat. Due to its wide range of phytotoxicity, it is considered as a potential pesticide. A study was conducted to explore the impact of BOA on the radicle and plumule elongation of mung bean (Phaseolus aureus) and associated changes in the macromolecular content - proteins and carbohydrates - and activities of enzymes like amylases, proteases, polyphenol oxidases and peroxidases. BOA significantly reduced the radicle and plumule length of P. aureus, and the contents of proteins and carbohydrates in both root and leaf tissue. On the other hand, activities of hydrolytic enzymes - proteases, amylases, polyphenol oxidases and peroxidases - increased substantially in both root and leaf tissue of P. aureus upon BOA exposure. This indicated that BOA treatment induced stress in P. aureus and enhanced enzyme activities to counter the induced stress and continue the growth. In other words, BOA-induced stress altered the plant biochemical status and related enzyme activities resulting in increased metabolism that serves to provide protection against cellular injury. Such studies providing information about the biomolecular content and enzymatic activities in response to natural products serve as clues for furtherance of knowledge about the modes of action of natural compounds of commercial interest. PMID- 17137119 TI - Biological activity of new N-oxides of tertiary amines. AB - Potential biological properties of newly synthesized single and double alkyl chain N-oxides of tertiary amines (NTA) were studied. Individual compounds in each of the series had alkyl chains of different length. Various experiments were performed to determine a mechanism of the interaction between NTA and model and biological membranes. These were measurements of hemolytic efficiencies of NTA (pig erythrocytes), their influence on the transition temperatures (DPPC liposomes), on potassium leakage from cucumber, its growth and chlorophyll content (Cucumis sativus cv. Krak F1), and on the resting membrane potential in alga cells (Nitellopsis obtusa). Also, prevention of erythrocyte membrane lipid oxidation induced by UV irradiation was studied. Potential antioxidative properties of NTA were additionally tested in radical chromogen (ABTS) experiments in which antioxidative efficiencies of NTA were compared to that of the standard antioxidant Trolox. It was found that NTA readily interacted with erythrocyte membranes. Their hemolyzing efficiency increased with the alkyl chain length. Slightly more intensive interaction was found for double alkyl chain compounds. Similar results were obtained in DSC experiments, where incorporation of NTA into liposomal membranes shifted the main transition temperatures and caused a broadening of the main transition peaks depending on the alkyl chain length. Double alkyl chain compounds were also found more efficiently influencing the growth of cucumber. Influence of NTA on the resting membrane potential of algae cells was not quite following the alkyl chain length rule found in erythrocyte and liposome experiments. Also potassium leakage and chlorophyll content determined in physiological experiments were not following the increase of lipophilicity of compounds. Most efficiently influencing those parameters were NTA having shorter alkyl chains, and efficiencies of single alkyl chain compounds were evidently stronger. Both methods used to test the antioxidative properties of NTA showed that they depended on the alkyl chain lengths of compounds within each series, but double alkyl chain ones exhibited markedly greater efficiency. PMID- 17137120 TI - Solid state biosurfactant production in a fixed-bed column bioreactor. AB - Biosurfactants are surface active substances which reduce interfacial tension and are produced or excreted at the microbial cell surface. We evaluated the biosurfactant production by Aspergillus fumigatus and Phialemonium sp. in solid state processes using fixed-bed column reactors. We evaluated two media, rice husks alone (simple support) and rice husks plus defatted rice bran (complex support), both enriched with either soy oil or diesel oil. The highest water-in oil emulsifying activity (EAw/o) obtained was 7.36 EU g(-1) produced by A. fumigatus growing on complex support enriched with soy oil and supplied with air at a rate of 60 mL g(-1) h(-1), while Phialemonium sp. had a maximum production of 6.11 EU g(-1) using the simple support with diesel oil and an aeration rate of 120 mL g(-1) h(-1). The highest oil-in-water emulsifying activity (EAo/w) was 12.21 EU g(-1) produced by Phialemonium sp. on the complex support enriched with diesel oil and at an aeration rate of 60 mL g(-1) h(-1), while A. fumigatus produced a maximum EAo/w of 10.98 EU g(-1) when growing on the complex support with no additional carbon source and an aeration rate of 60 mL g(-1) 1 h(-1). PMID- 17137121 TI - Surfactin isoforms from Bacillus coagulans. AB - Bacillus coagulans has been found to produce several surfactins that are powerful lipopeptide surfactants. Four main components with molecular weights 1007, 1021 and 1035 Da were separated. Their structures have been confirmed by spectrometric and spectroscopic studies and by acid hydrolysis. The compounds were found to represent two pairs of surfactin isoforms in which beta-hydroxy-iso-C14 or anteiso-C15 fatty acids are linked to the [Leu7] or [Val7] heptapeptide moiety by both an amide group and a lactone bond. PMID- 17137122 TI - Non-invasive monitoring of the light-induced cyclic photosynthetic electron flow during cold hardening in wheat leaves. AB - The effect of irradiance during low temperature hardening was studied in a winter wheat variety. Ten-day-old winter wheat plants were cold-hardened at 5 degrees C for 11 days under light (250 micromol m(-2) S(-1)) or dark (20 micromol m(-2) s( 1)) conditions. The effectiveness of hardening was significantly lower in the dark, in spite of a slight decrease in the Fv/Fm chlorophyll fluorescence induction parameter, indicating the occurrence of photoinhibition during the hardening period in the light. Hardening in the light caused a downshift in the far-red induced AG (afterglow) thermoluminescence band. The faster dark re reduction of P700+, monitored by 820-nm absorbance, could also be observed in these plants. These results suggest that the induction of cyclic photosynthetic electron flow may also contribute to the advantage of frost hardening under light conditions in wheat plants. PMID- 17137123 TI - Spectroscopic characterization of the artificial Siderophore pyridinochelin. AB - Siderophores play a very important role in the uptake process of iron by bacteria. Due to the so-called active transport the uptake of siderophores by bacteria is very specific, which makes the use of siderophores as effective shuttles for antibiotics in the treatment of infections and other diseases caused by bacteria highly attractive. In order to further investigate the transport and incorporation of siderophores into the bacteria cells, distinct molecular probes are needed. Especially artificial siderophores, that show a specific intrinsic fluorescence, are highly attractive for such monitoring purposes. A promising candidate of such a fluorescent artificial siderophore is bis-2,3 dihydroxybenzoyl-2,6-dimethylamino-pyridine (pyridinochelin, PY). The fluorescence properties of PY were investigated in different solvents and in the presence of different metal ions. It was found that PY in its free form shows a complex fluorescence behavior. In methanol a clear dual fluorescence is observed. In aqueous solution intermolecular interactions with water molecules are determining the intrinsic fluorescence. Upon complexation with metal ions (Me3+ = Eu3+, Tb3+, Al3+, Fe3+) the fluorescence characteristics changed. The fluorescence quantum yield of PY decreased upon addition of Me3+--except for Al3+, which showed no fluorescence quenching. The fluorescence decay of PY loaded with metal ions showed a nicely mono-exponential fluorescence decay, which was in contrast to PY in the absence of metal ions. This drastic change in the fluorescence properties of PY upon metal ion complexation makes PY highly attractive as a fluorescence probe for the investigation of siderophore action and siderophore-mediated transport processes. PMID- 17137124 TI - Electrophysiological responses of Atta sexdens rubropilosa workers to essential oils of eucalyptus and its chemical composition. AB - The leaf-cutting ant Atta sexdens rubropilosa Forel, 1908 is the most harmful of the Eucalyptus pests, causing severe losses in wood production through defoliation. Various strategies have been tried and effort spent on the development of methods to control this pest, however no practical and environmentally acceptable one currently exists. In this work the chemical composition of the essential oil of seven Eucalyptus species was identified and the selectivity and sensitivity of antennal receptors of A. sexdens rubropilosa workers to the volatile compounds were determined using the electroantennographic technique (EAG and GC-EAD). Analysis by GC-EAD showed in E. cloesiana and E. maculata, respectively, seventeen and sixteen terpenes that elicited responses in ant workers' antennae, indicating the potential role of the essential oils as allelochemicals that determine the choice of the foraging material. PMID- 17137125 TI - Effect of alloxan-diabetes and subsequent treatment with insulin on kinetic properties of succinate oxidase activity from rat liver mitochondria. AB - We evaluated early and late effects of alloxan-diabetes and subsequent insulin treatment on the kinetic properties of succinate oxidase (SO) in rat liver mitochondria. Diabetic state lowered the SO activity; insulin treatment was effective in restoring the activity only in one-week diabetic rats. The energies of activation in low and high temperature ranges (EH and EL) decreased significantly in diabetic animals; once again insulin treatment was partially effective only in the one-week diabetic group. The total phospholipids (TPL) and cholesterol (CHL) contents did not change in one-week groups. In one-month diabetic animals TPL decreased while CHL increased; insulin treatment induced further changes without restoring normality. The lysophospholipid (Lyso), sphingomyelin (SPM), phosphatidylinositol (PI) and phosphatidylserine (PS) content increased in the diabetic state while phosphatidylcholine (PC) and phosphatidylethanolamine (PE) decreased. Insulin treatment had a partial restorative effect. The changes in EH correlated negatively with SPM. The phase transition temperature, Tt, decreased in diabetic and insulin-treated groups. These changes correlated positively with the ratios of TPL/PI and TPL/PS. The membrane fluidity decreased in the diabetic state; insulin had a restorative effect only in the one-week group. PMID- 17137126 TI - Lipids in the femoral gland secretions of male Schreiber's green lizards, Lacerta schreiberi. AB - In spite of the importance of chemoreception and chemical signals in social organization of lizards, only a few studies have examined the chemical composition of secretions that lizards use for intraspecific communication. The secretion of the femoral glands of male Schreiber's green lizards (Lacerta schreiberi) contains 51 lipophilic compounds, including several steroids, a tocopherol, n-C9 to n-C22 carboxylic acids and their esteres, and minor components such as alcohols between C12 and C24, two lactones, two ketones, and squalene. These compounds were identified on the basis of mass spectra, obtained by GC-MS. We compared these chemicals with those found in other lizard species, and discussed how environmental conditions could explain the differential presence of chemicals in different lizards. Particularly, the high abundance of a tocopherol in this lizard is suggested to contribute to avoid oxidation of other lipids in secretions, increasing chemical stability of scent marks in the humid conditions of its habitat. PMID- 17137127 TI - Sophoraflavanone G from sophora pachycarpa enhanced the antibacterial activity of gentamycin against Staphylococcus aureus. AB - In this study the enhancement effect of Sophora pachycarpa roots' acetone extract on the antibacterial activity of gentamycin was evaluated against Staphylococcus aureus. Disc diffusion and broth dilution methods were used to determine the antibacterial activity of gentamycin in the absence and presence of plant extract and its various fractions separated by TLC. A clinical isolate of S. aureus was used as test strain. The active component of the plant extract involved in enhancement of gentamycin's activity had Rf = 0.72 on a TLC plate. The spectral data (1H NMR, 13C NMR) of this compound revealed that this compound was 5,7,2',4' tetrahydroxy-8-lavandulylflavanone (sophoraflavanone G), previously isolated from Sophora exigua. In the presence of 0.03 microg/ mL of sophoraflavanone G the MIC value of gentamycin for S. aureus decreased from 32 to 8 microg/mL (a four-fold decrease). These results signify that the ultra-low concentration of sophoraflavanone G potentiates the antimicrobial action of gentamycin suggesting a possible utilization of this compound in combination therapy against Staphylococcus aureus. PMID- 17137128 TI - Application of ultrasonography in the study of the reproductive system of tropical jennies (Equus asinus). AB - The use of high-frequency (5 MHz) ultrasonography was studied in 11 jennies (7 non-pregnant and 4 pregnant) to characterize the reproductive organs and follicular activities at different stages of reproduction. The result showed close similarity with the mare. The visibility of endometrial folds increased towards ovulation. A positive correlation (p < 0.001; tau = 0.79) was found between the score of the folds and the size of the dominant follicle. The diameter of the uterus and the size of the dominant follicle were significantly correlated (p < 0.001; r = 0.80). In pregnant jennies, an embryonic vesicle was detectable at 14 days. Follicular growth was characterized by more than one wave. The smallest ovarian follicle was 2 mm and the largest 40mm. Depending on the reproductive stage, up to 13 follicles were detected per ovary. After monitoring 84 cycles, a mean (+/- SD) diameter of 34.4 +/- 3.6 mm (27.5-40.2 mm) of the preovulatory follicle and 67.85% incidence of single ovulation were found. The mean (+/-SD) interovulatory interval was 25.7 +/- 6 days. This study proved that high-frequency ultrasonography is highly effective in characterizing the reproductive organs and follicular activity of jennies and could be useful in the reproductive management of donkeys. PMID- 17137129 TI - Responses to tuberculin among Zebu cattle in the transhumance regions of Karamoja and Nakasongola district of Uganda. AB - Responses to tuberculin in Zebu cattle of the transhumant pastoral farming system in Karamoja region and Nakasongola district in the north-eastern and mid-central regions in Uganda, respectively, were investigated using a comparative intradermal tuberculin skin test. Of the 1864 cattle tested from 30 large units (superherds) in Karamoja and 7 herds in Nakasongola, a total of 28 animals from 19 herds (51.4%) tested positive. Inter-district tuberculin reactor prevalence variations seemed to be influenced by climate, with impact on both the management patterns and transmissibility of agent. High herd tuberculin reactor prevalence (51.4%) was attributed to widespread contacts and mixing of animals between herds. Low individual animal tuberculin test positivity (mean = 1.4%) was attributed to low transmissibility of the agent under the Karamoja climate, which is semi-arid, and to increased resistance due to non-specific response to environmental mycobacteria and natural selection, since there was no active control against bovine tuberculosis. Owing to similarities in management practices in Karamoja and widespread risk factors, it was difficult to identify which were more important, but variations in sources of drinking water pointed to provision of lake and borehole water during dry season as reducing the risk. Positive bovine tuberculin reactor prevalence and skin reactor status were related to age. PMID- 17137130 TI - A preliminary study of the role of ducks in the transmission of Newcastle disease virus to in-contact rural free-range chickens. AB - The role of ducks in the transmission of Newcastle disease virus (NDV) to free range village chicken was investigated experimentally. Newcastle disease (ND) seronegative ducklings reared in a pen were infected oronasally with velogenic NDV of intracerebral pathogenicity index (ICPI) 1.8 isolated from outbreaks in village chickens in Uganda. A first group of 3-week-old ND seronegative chicks was mixed with the ducks and they were kept together for 7 days. Both ducks and chicks were observed for ND clinical signs and any mortality, and they were bled and their sera were tested for ND antibodies by haemagglutination inhibition (HI) test. The chicks were removed, euthanized and examined for any ND lesions, while the ducks were transferred to a fresh pen and a second group of chicks was introduced and observed and treated as above. The ducks and the chicks tested positive for ND antibodies 7 days post infection and contact, respectively, but showed no clinical signs, post-mortem lesions or mortality. The mean ND antibody titre of the second group of chicks was lower than for the first group. This study has shown that although ducks can be infected with velogenic NDV, they do not show clinical signs but are able to transmit NDV to in-contact chicks. Further investigations are needed of the lack of clinical signs in the in-contact chicks and how long the ducks remain infective. PMID- 17137131 TI - Estimated direct economic costs associated with tick-borne diseases on cattle in Tanzania. AB - Tick-borne diseases, namely, anaplasmosis, babesiosis, cowdriosis and theileriosis, constrain cattle production and improvement in Tanzania, leading to considerable economic losses. A simple spreadsheet model was used to estimate the economic losses resulting from production losses, treatment and control costs associated with tick-borne diseases (TBD) in Tanzania. Model parameters included the national cattle population, reported TBD morbidity, fatality risk, and chemotherapy and control measures used. The total annual national loss due TBD was estimated to be 364 million USD, including an estimated mortality of 1.3 million cattle. Theileriosis accounted for 68% of the total loss, while anaplasmosis and babesiosis each accounted for 13% and cowdriosis accounted for 6% of the total loss. Costs associated with mortality, chemotherapy and acaricide application accounted for 49%, 21% and 14% of the total estimated annual TBD losses, respectively, infection and treatment method milk loss and weight loss accounted for 1%, 6% and 9% of the total annual loss, respectively. Despite the inadequacies of the data used, the results give evidence that tick-borne diseases inflict substantial economic losses on cattle production and resource use in Tanzania. PMID- 17137132 TI - Prevalence of Sarcocystis in slaughtered one-humped camels (Camelus dromedarius) in Iran. PMID- 17137133 TI - Identification and quantification of subsidies relevant to the production of local and imported pig breeds in Vietnam. AB - Livestock diversity contributes in many ways to human survival and well-being, while its loss reduces options for attaining sustainable agriculture and universal food security. The current rapid rate of loss of this diversity is the result of a number of underlying factors. While in some cases changes in production systems and consumer preferences reflect the natural evolution of developing economies and markets, in other cases production systems, breed choice and consumer preferences have been distorted by local, national and international policy. In the context of a widespread threat to local pig breeds in Vietnam, this paper identifies and quantifies the level of agricultural subsidies that are currently contributing to this process of breed substitution. Producer subsidies which tend to improve the competitiveness of imported breeds and their crosses over local breeds--are shown to be considerable, and mitigating measures are now urgently needed to avoid an irreversible loss of livestock diversity. PMID- 17137134 TI - Effect of supplementation on the feed intake and performance of confined and scavenging crossbred growing chickens in Burkina Faso. AB - An experiment was conducted to evaluate the performance of crossbred growing chickens (Rhode Island Red x indigenous Burkina Faso hens) from 6 to 17 weeks of age, under five feeding/management regimes: (1) CMx(+), confined and given a mixed feed containing cracked maize and cowpea and a vitamin-mineral premix ad libitum; (2) CS(+), confined and offered ad libitum a choice of cracked maize and cowpea with the premix; (3) ScS(+), scavenging from 09:00 to 16:00 with the diet in treatment (2) available from 16:00 to 09:00; (4) ScS(-), treatment (3) but without the premix; and (5) ScO, scavenging only, with no supplements provided. Daily dry matter (DM) intake was highest for CS(+) (43.5 g), and lowest for CMx(+) (33.6 g) (p < 0.05), with intermediate intakes for ScS(+) and Sc(-) of 36.7 g and 36.2 g, respectively. The ratios of intakes of cowpea to maize were 50:50, 21:79, 27:73 and 22:78 for CMx(+), CS(+), ScS(+) and ScS(-), respectively (p < 0.05). Dietary concentrations of crude protein (CP) were 15.7%, 11.5%, 12.3% and 11.6% of DM for CMx(+), CS(+), ScS(+) and ScS(-), respectively. Average daily gains (ADG) were 8.15 g, 5.24 g, 6.03 g, 5.36 g and 4.45 g for CMx(+), CS(+), ScS(+), ScS(-) and ScO, respectively, and were significantly higher for CMx(+) (p < 0.05). Feed conversion ratio was highest for CS(+) and lowest for CMx(+). ADG of the males (6.44 g) was significantly (p < 0.05) higher than that of the females (5.86 g). Breast and thigh muscle weights were highest for ScS(+) (p < 0.05). PMID- 17137135 TI - Reproductive performances of dairy cows in smallholder production system in Selalle, Central Ethiopia. AB - A study was conducted to assess the reproductive performance and factors that influence reproductive efficiency of cows in smallholder dairy farms in Selalle, Central Ethiopia. Three hundred dairy farms (average herd size = 8) were visited and data on reproductive, breeding and management histories were collected and analysed. The overall geometric means for calving to conception interval (n=382) and duration after last calving (n=422) were 187 and 201 days in pregnant and non pregnant cows, respectively. The least-squares mean calving to conception interval was higher (p < 0.05) in mixed crop-livestock production (MCLP) than in small urban dairy production (SUDP) systems and was lower (p < 0.01) in non suckling than in suckling cows. District significantly influenced (p < 0.01) the least-squares mean duration after last calving and, among the districts, non pregnant cows in Wuchale-Jida had the highest values, whereas cows in Sululta and Mulo had the lowest values. The overall average number of services per conception (+/-SD) and the first service conception rate were 1.6 +/- 1.0 (n=382) and 56% (n=456), respectively. The prevalences of abortion, dystocia (assisted parturition), retained fetal membrane, vulval discharge/endometritis and pre weaning calf mortality were 1.4%, 1.3%, 5.4%, 2.8% and 17.4%, respectively. The present estimates of extended calving to conception interval and duration after last calving indicate poor reproductive performances of cows in Selalle smallholder dairy farms. The pre-weaning calf mortality rate is highly significant. Accordingly, a further detailed investigation is necessary to identify and quantify the specific reproductive disorders and associated interacting factors attributing to such poor performance and to determine the causes and predisposing factors behind such high calf mortality. PMID- 17137136 TI - Effect of feeding different levels of foliage from Cratylia argentea to creole dairy cows on intake, digestibility, milk production and milk composition. AB - The effect of feeding different levels of foliage from Cratylia argentea (Desvaux) O. Kuntze to dairy cows on intake, digestibility, milk production and milk composition was studied in the dry tropics in Nicaragua. The treatments were sorghum silage ad libitum, either unsupplemented or supplemented with 2 kg or 3 kg of Cratylia on a dry matter (DM) basis. Six Bos indicus cows of the Creole Reyna breed, with a mean body weight of 386 (SD 19) kg were used in a replicated 3 x 3 Latin square design. Supplementation with Cratylia increased (p < 0.05) DM intake from 6.6 to 7.8 and 8.7 kg DM/day and milk production from 3.9 to 5.1 and 5.7 kg/day for sorghum silage alone and supplementation with 2 kg and 3 kg DM of Cratylia, respectively. Milk fat, total solids and crude protein and organoleptic characteristics (smell, taste and colour) were not significantly different among diets. The apparent digestibility coefficients of dry matter, organic matter, neutral detergent fibre and acid detergent fibre were not affected significantly by supplementation with Cratylia. However, crude protein (CP) digestibility increased (p < 0.05) in the diets supplemented with Cratylia compared to sorghum silage alone. In conclusion Cratylia given as a protein supplement to a low quality diet improved DM intake and CP digestibility of the diet and increased milk production, but did not affect milk composition. PMID- 17137137 TI - Monthly variation of fertility and oestrus frequency in crossbred dual-purpose cows in three agroecological areas of the South American tropics. AB - The aim of this study was to examine the monthly variation of the first service conception (FSC) and oestrus frequency (OF) from 8308 artificial inseminations (AI) of 2960 crossbred dual-purpose cows on three commercial farms with improved management located in the Maracaibo Lake Basin, Venezuela. The effects of month of the year, agroecological area (from dry tropical forest to sub-humid tropical forest on the three farms), predominant breed (BT, Bos taurus; BI, Bos indicus), and season (December-April; May-August; and September-November) were considered. Data were analysed using logistic regression (FSC) and GLM (OF) from SAS. The mean FSC was 55.4% overall and 48.7%, 57.0% and 58.3% for farms A, B and C, respectively (p < 0.01). Within the three agroecological areas (farms A, B and C), the FSC was highest during the cooler and drier months of the year (season 1), while the lowest FSC was obtained during the months of highest rainfall and humidity (season 3) (53.3% vs. 37.4%; 58.5% vs. 49.0% and 63.0% vs. 52.3% on farms A, B, C, respectively; p < 0.01). In the three studied farms, OF was higher during the first 4 months of the year; with a diminishing trend towards the end of the year (p < 0.05). In general, BI cows had a higher FSC than BT cows, especially during the second half of the year, when environmental conditions were characterized by higher rainfall and humidity with lower wind velocity (58.8%a, 55.5%b and 46.4%c in season 1, 2 and 3 respectively; a,b,c p < 0.01). PMID- 17137138 TI - Reproductive response in suckling Corriedale ewes to the ram effect during the non-breeding season: effect of postpartum condition and the use of medroxyprogesterone priming. AB - The response to the ram effect and the use of progestagen priming was studied in postpartum suckling Corriedale ewes. Two experiments were carried out during the non-breeding season. In experiment 1, the effectiveness of the ram effect to induce fertile oestrus in postpartum ewes was determined. While no significant differences in the frequency of ewes in oestrus were found, the overall conception rate was significantly higher in ewes that were weaned at least 6 months before the study started (63.3%) than in 60-90 days postpartum ewes (45.3%, p < 0.01). In experiment 2, the influence of medroxyprogesterone acetate priming on the response to the ram effect in suckling Corriedale ewes was determined. The frequency of ewes in oestrus and conception rate between ewes that remained unprimed and 6-days-primed ewes were similar. We conclude that it is possible to induce oestrus in postpartum suckling Corriedale ewes using the ram effect during the non-breeding season and that the use of progestagen priming seems to be ineffective in improving the response of postpartum ewes to the ram effect. PMID- 17137139 TI - The relationship between serum creatinine and estimated glomerular filtration rate: implications for clinical practice. AB - INTRODUCTION: A new classification of chronic kidney disease (CKD) has been widely adopted that stratifies patients into 5 'stages' according to estimated glomerular filtration rate (eGFR). In adults the most commonly used formulae to calculate eGFR are the Cockcroft and Gault (C and G) and Modification of Diet in Renal Disease (MDRD) formulae. The UK Renal Association has recommended calculation of MDRD eGFR to screen for reduced kidney function in primary and secondary care. AIM: The aim of this study was to explore the implication of using these predictive formulae. METHODS: We searched for patients currently attending a renal clinic who have ever had a serum creatinine (SCr) of exactly 100 micromol/L, 150 micromol/L or 200 micromol/L. The C and G and MDRD eGFRs corresponding to that SCr were calculated. The proportion of patients in each stage of the CKD classification was determined. RESULTS: For a SCr of 100 micromol/L mean eGFR was 86.5 ml/min (range 31.0 - 192.8) by C and G and 63.8 ml/min (range 39.7 - 99.9) by MDRD (p < 0.0001; t-test of mean). For SCr 150 micromol/L mean eGFR was 51.7 ml/min (18.0 - 110.4) by C and G and 38.0 ml/min (20.7 - 54.8) by MDRD (p < 0.0001). For SCr of 200 micromol/L mean eGFR was 34.4 ml/min (12.6 - 89.5) by C and G and 27.3 ml/min (16.7 - 41.3) by MDRD (p < 0.0001). Using MDRD eGFR 46.5% patients with a SCr of 100 micromol/L have stage 3 CKD (GFR 30-60 ml/min) and all patients with a SCr of 150 micromol/L or 200 micromol/L have CKD 3 or worse. 8.6% of males with SCr 100 micromol/L had stage 3 CKD or worse compared with 86.8% females. 70.2% patients > 65 years old with SCr 100 micromol/L had stage 3 CKD. CONCLUSIONS: Targeted screening of patients at risk for CKD will identify a large number of patients who require management of CKD and potential referral to nephrology services even at levels of SCr regarded as 'normal' or mildly. PMID- 17137140 TI - Management of COPD in primary care in north-east Scotland. AB - INTRODUCTION: We wished to obtain a snapshot of current service provision and how this could best be developed approximately one year on from the introduction of the National Institute for Clinical Excellence (NICE) guidelines for the management of chronic obstructive pulmonary disease (COPD) and the inclusion of COPD care in the New GMS Contract Quality and Outcomes Framework (QOF). METHODOLOGY: A questionnaire-based survey sent to every general practice (n = 84) in Grampian. RESULTS: Responses were received from 75 of 84 practices (89%). Questionnaires were returned by both general practitioners (GPs) and practice nurses in 45 practices (54%). All responding practices reported that they had COPD registers. 60/75 (80%) of practices reported having a dedicated COPD clinic; 70/75 (93%) had a spirometer. Areas identified for service development were: quality assuring training in COPD care and spirometry; expanding pulmonary rehabilitation provision (86%), delivering this service locally (54%) and in primary care (75%); standardising referral, assessment and communication about provision of home oxygen; training in pulse oximetry (71%). CONCLUSION: This data has important implications for the validity of the quality indicators (QOF) under the new GMS contract. Our respondents identified areas where the new GMS contract QOF could be improved, as well as providing useful suggestions for service development. Respondents recognised that not all clinical services can be effectively delivered by general practice with data supporting the development of intermediate care services for people with COPD. PMID- 17137141 TI - Comparative evaluation of laparoscopic versus open nephrectomy in children. AB - BACKGROUND AND AIMS: To assess the benefits of laparoscopic nephrectomy in children as compared to open surgery and to ascertain whether or not the retroperitoneal technique offers any surgical advantage over the transperitoneal approach. METHODS AND RESULTS: This study gathered retrospective data on a consecutive series of 51 paediatric patients comparing; operative time, length of hospital stay and analgesic requirement. Results show that when laparoscopic surgery is compared with the open technique it took 25 mins longer to perform, there was a shorter post operative stay by 2.5 days, and a 50% reduction in morphine use. Patients who had the retroperitoneal technique were discharged from hospital within one day. CONCLUSIONS: Compared to previous studies, the findings in this study suggest that there are distinct benefits in laparoscopy for children. There is some evidence in favour of the retroperitoneal technique. Patients are in hospital for shorter periods and need less analgesia. PMID- 17137142 TI - Outcome using a conservative tracheostomy strategy in intensive care. AB - BACKGROUND AND AIMS: We have a conservative approach to tracheostomy in intensive care, in that every patient considered for tracheostomy in our unit is discussed at a clinical meeting to evaluate potential benefit and harm from the procedure. This study examined tracheostomy numbers, complications and outcome of patients who had tracheostomy in comparison to our general intensive care population and to Scottish national data. METHODS: We collected prospective data on all patients having a tracheostomy over a three year period. Data included included time to tracheostomy, type of procedure (open or percutaneous), ventilator settings immediately before tracheostomy, peri- and post-operative complications and mortality. RESULTS: The number of tracheostomies was low (27 of 692 patients, 4%) and time to tracheostomy high (mean, 18 days) in comparison to published data. Patients who had a tracheostomy had a hospital mortality that was greater than predicted by the APACHE II scoring system (55% actual mortality vs. 44% predicted). Standardised mortality ratio (SMR) was 1.29 for tracheostomy patients and 0.89 for patients who did not have a tracheostomy. Length of stay and mortality were similar to national figures. One patient in the tracheostomy group developed tracheal stenosis that needed surgical intervention. There were no cases of symptomatic tracheal stenosis in the much larger group managed with orotracheal intubation alone. CONCLUSIONS: We found that a conservative approach to performing tracheostomy reduced the number of procedures performed without affecting overall unit outcomes. PMID- 17137143 TI - The prevalence of alcohol misuse among acute admissions: current experience and historical comparisons. AB - BACKGROUND: Over the last 25 years there has been a large increase in alcohol related deaths in Scotland. Medical patients who misuse alcohol may have overt alcohol related disease, but may also present with other unrelated illness. AIM: We examined alcohol misuse amongst acute medical admissions to compare this with other similar studies at the same hospital since 1974. PATIENTS AND METHODS: 850 consecutive admissions to the medical receiving unit of Victoria Infirmary were assessed. They were assessed using the modified Michigan Alcohol Screening Test (MAST) and also by a medical consultant. 414 patients also had their blood ethanol levels measured on admission. RESULTS: 18.6% admissions had a MAST greater than 5 and were considered to misuse alcohol (24.8% male and 12.2% female; p < 0.0001). Patients from socio-economic group V and patients presenting with gastro-intestinal haemorrhage or self-poisoning were more likely to misuse alcohol. The sensitivity and specificity of consultant opinion regarding alcohol misuse were 0.55 and 0.97 compared with the MAST. There was an increase in the alcohol misuse amongst women admitted (12.2%) compared to 1977 (5.5%; p = 0.0026) and 1981/2 (6.3%; p = 0.004). CONCLUSION: Alcohol misuse is common amongst acute medical admissions. Since 1979, there has been a particular increase in female medical admissions who misuse alcohol. Medical opinion regarding alcohol misuse lacks sensitivity in identifying at risk individuals compared with a validated. PMID- 17137144 TI - Urgent in-patient coronary angiography: a comparison of centres with and without cardiac catheter facilities. AB - OBJECTIVES: To review the referral of patients to a tertiary centre for urgent angiography and to determine if there are differences in invasive treatment strategies for patients with acute coronary syndrome (ACS). METHODS: There were 2 parts to the study, a retrospective part over 3.5 years from a computerised cardiac laboratory booking data base and a prospective part over 3 months. RESULTS: There were 1190 urgent in-patient angiograms performed with 499 (42%) admitted initially to the tertiary centre while the remaining 691 (58%) were admitted to district general hospitals (DGH), with no on-site access to a cardiac laboratory, and subsequently transferred to the tertiary centre. Once referred, DGH patients waited longer for their angiogram (2.7 +/- 3.2 vs 2.0 +/- 2.8 days, p < 0.0001). Interestingly, DGH patients appear to spend an average of 4 days in hospital prior to referral for angiography. DGH patients were more likely to have a higher Thrombosis in Myocardial Infarction (TIMI) risk score at presentation and following angiography were more likely to have coronary artery bypass graft (CABG) or percutaneous coronary intervention (PCI) and less likely to have angiographically normal arteries. CONCLUSIONS: Our findings are consistent with previous studies demonstrating that access to coronary angiography varies considerably between hospitals. However, we have demonstrated that patients in DGHs wait on average 4 days before referral for coronary angiography suggesting that there may be triage based on initial responses to medical therapy. Further research is needed to determine whether this has a direct effect on outcomes. PMID- 17137145 TI - Are medical students adequately trained to prescribe at the point of graduation? Views of first year foundation doctors. AB - BACKGROUND: Drugs are the major therapeutic intervention provided by most doctors throughout their careers. The General Medical Council expects all medical students to be competent to prescribe at the point of graduation. AIMS: The aim of this study was to assess the views of Foundation Year 1 (FY1) doctors who had recently graduated from the University of Edinburgh about their training and competence in relation to the use of drugs based on their early clinical experience. METHOD: A questionnaire was constructed based on Tomorrow's Doctors 2002 and distributed to FY1 doctors who graduated in August 2005. RESULTS: Responses were received from 100 (39.8%) of the doctors who graduated in 2005. Only 32% respondents considered themselves 'competent to prescribe' at the point of graduation. Less than 50% of respondents felt comfortable in providing information about possible treatments to allow patients to make informed decisions about their care. The majority of respondents complained about a lack of formal teaching and practice at basic clinical skills relating to drug therapy. CONCLUSION: Many graduates feel under-prepared to take on prescribing responsibilities after graduation. These findings emphasise the need to ensure that all medical curricula are able to provide sufficient learning opportunities and robust assessment in this important area of clinical practice. PMID- 17137146 TI - Rate of referral of breech infants for hip ultrasound: an audit cycle. AB - BACKGROUND: Infants with risk factors for developmental dysplasia of the hip (DDH) should have hip ultrasounds performed shortly after birth to detect and treat the condition at an early stage. Breech presentation is associated with increased risk of DDH. We embarked on an audit cycle to assess and improve our rate of referral of breech infants for hip ultrasound. METHODS: Two retrospective audits (phases I and II) were carried out before, and after, the introduction of a new pro-forma which encouraged recognition of breech presentation at the time of the routine neonatal examination. Breech infants were identified from labour ward records. Multiple births and infants < 35 weeks gestation were excluded. Infants were considered to have been referred for ultrasound if the computer system at our affiliated children's hospital held a record of an ultrasound appointment. RESULTS AND CONCLUSIONS: In phase I 56% of breech infants born in our hospital had been referred for hip ultrasound. In phase II the referral rate had risen to 76% (p = 0.034). We conclude that the change in practice was effective. Further improvement might be achieved by increasing staff awareness of risk factors for DDH and by enlisting the help of advanced neonatal nurse practitioners (ANNPs) in routine neonatal examinations. PMID- 17137147 TI - Patients' attitudes towards mydriasis for diabetic eye disease screening. AB - AIMS: The purpose of the questionnaire was to explore attitudes of patients towards mydriasis for diabetic retinopathy screening. METHODS: Two groups of patients were invited to take part: group 1 comprised patients attending the diabetic clinic and had previous experience of mydriasis, group 2 patients attending the mobile screening unit for non-mydriatic digital retinal photograph and who were not previously used to receiving mydriatic eye drops. Basic demographic data was recorded and the volunteers invited to complete a questionnaire. RESULTS: 292 patients were recruited into group 1 (median age 63 years range 20-94) and 103 into group 2 (median age 68 years range 29-96). 42% of patients in group 2 indicated that they were unhappy with the use of dilating eye drops and 26% of this group reported that they may be discouraged from attending screening for diabetic retinopathy if drops were introduced. These figures were statistically lower in group 1 at 8% and 1-8% respectively (p < 0.001). Blurring of vision was identified as the most troublesome feature of the use of mydriasis for patients. A large proportion of patients drove themselves to their last screening appointment; 58% in group 2 and 39% in group 1. A third of all patients (33%) indicated that they worked outside the home. CONCLUSIONS: Many patients attending diabetic eye screening return to driving and work immediately after the appointment. Introduction of the use of routine drops may discourage attendance. Education and experience may have an important role in improving acceptability of mydriatic eye drops. Retinal screeners need to have clear guidelines with which to advise patients. PMID- 17137148 TI - Further observations on the removal of an enormous facial tumour by Robert Liston in 1834. AB - In 1834, shortly before Robert Liston (1794-1847) left Edinburgh to take up the post of senior surgeon to the North London (now University College) Hospital, he operated on Mrs Fraser, from Banchory, in north-east Scotland. Her tumour was believed to have originated as a result of trauma to the left maxillary antrum. It was immense in size when he first saw it, and produced an enormous degree of facial distortion. The tumour was associated with drooping and disfigurement of the left angle of her mouth and extended forwards from her left external ear to the left side of her nose. Its upper part eventually obstructed the vision of her left eye, while its lower part extended for some inches below the level of her mandible. The volume of the tumour was just slightly less than that of her face. Liston provided a detailed description of her appearance when he first saw her, and gave a detailed history that suggested its possible aetiology. Descriptions of her pre-operative cast were previously published in this Journal in 2000. Since then, additional casts showing her post-operative appearance and that of her tumour have been located, and these form the basis for this follow-up account. PMID- 17137149 TI - Clarity on operating theatre design. PMID- 17137150 TI - Support service issues scrutinised. PMID- 17137151 TI - The missing Health Technical Memoranda. PMID- 17137152 TI - HCAI reduction needs high level direction. PMID- 17137153 TI - Exemplar status for NHS trust. PMID- 17137154 TI - Tyneside Trust goes green. PMID- 17137155 TI - NHS debt crisis forces a rethink. PMID- 17137157 TI - Off-site systems advance. PMID- 17137156 TI - Legionella precautions not to be taken lightly. PMID- 17137158 TI - Protecting sensitive patient data. PMID- 17137159 TI - Visual healing in healthcare. PMID- 17137160 TI - Providing a five-star service. PMID- 17137161 TI - Pure water technology aids decontamination. PMID- 17137162 TI - A safer window for vulnerable patients. PMID- 17137163 TI - Small outlays yield large benefits. PMID- 17137164 TI - Spotlight on hospital water controls. PMID- 17137165 TI - FMs need strong partners. PMID- 17137166 TI - [Prognostic risk factors for serious complications in an inpatient population with neutropenia at the onset of a febrile episode]. AB - Patients with neutropenia and fever conform a heterogeneous population with a variable risk of serious complications and mortality. The goal of this study was to identify prognostic risk factors present at the beginning of the episode, for adverse events and serious complications in patients admitted in a general ward with fever and neutropenia. A cohort of 238 episodes with neutropenia and fever (neutrophils < 1000/mm3 and T > 38.3 00) in 167 patients admitted to our general hospital between 1997 and 2004 was followed. Eighty two percent of the patients had hematologic malignancies, 14% solid tumors and 4% were not associated with chemotherapy. Sixty seven adverse events were registered (46% renal insufficiency, 27% refractory hypotension, 15% respiratory insufficiency and 12% major bleeding). Significant differences were found in presence of current co morbidities, body temperature > 39 00, heart rate > 120 beats per minute, respiratory rate > 24 per minute, systolic blood pressure < 90 mm Hg, presence of 3 or more altered laboratory values, presence of a clinical site of infection and positive blood cultures. The logistic regression multivariate analysis showed that the following characteristics were independently associated with adverse events: systolic blood pressure < 90 mm Hg (OR = 7, p < 0.01), current co morbidities (OR = 8.5, p = 0.02), respiratory rate > 24 per minute (OR = 2.8, p = 0.01), and the presence of a clinical site of infection (OR = 2.1, p = 0.03). The presence of systolic hypotension, high respiratory rate, current co-morbidities and a clinical site of infection at the time of admission were identified predictors of subsequent serious complications in patients admitted with fever and neutropenia in a general ward. PMID- 17137167 TI - [Echocardiographic analysis of the effect of different Na+/H+ exchanger inhibitors on left ventricular structure and function in spontaneously hypertensive rats]. AB - The aim of this study was to analyze by echocardiogram, the action of two Na+/H+ exchange, inhibitors, HOE 642 (HOE) and BIIB 723 (BIIB) on left ventricular (LV) mass and LV systolic function. We studied 16 spontaneously hypertensive rats (SHR), 8 treated with HOE 30 mg/kg/day, 8 with 30 mg/kg/day of BIIB during 30 days and 4 SHR as controls during those 30 days. Results are expressed as mean values +/- SEM. The systolic blood pressure and the echocardiograpic parameters examined did not evidence changes during that period in the controls rats. Even though HOE determined a slight decrease in blood pressure (HOE C: 184 +/- 1.75 mm Hg; HOE 30d: 176.20 +/- 2.60 mm Hg - p < 0.01) which was not detected with BIIB, both drugs provoked an increase of peak systolic stress (HOE C: 166 +/- 29 kdynes/cm2; HOE 30d: 204 +/- 34 kdynes/cm2, p < 0.04; BIIB C: 164 +/- 25.90 kdynes/ cm2; BIIB 30d: 234 +/- 29.30 kdynes/cm2, p < 0.02). HOE and BIIB reduced LV mass after 30 days of administration (HOE C: 612.50 +/- 50 mg; 30d: 452 +/- 37 mg, p < 0.01. BIIB C: 544 +/- 16mg; 30d: 374 +/- 25 mg, p < 0.01). LV endocardial shortening was similar independently of the NHE inhibitors used (HOE C: 62.30 +/- 2.75%; 30d: 65.50 +/- 2.40%, ns. BIIB C: 63.20 +/- 2,39%; 30d 67,20 +/- 1.62%, ns). These data demonstrate that long-treatment with HOE or BIIB produced similar LV mass regression without changes in endocardial fractional shortening in spite of the increase of peak systolic stress. This finding could represent an increased inotropism previously depressed by the development of hypertrophy. PMID- 17137168 TI - [Multidrug-resistant tuberculosis in AIDS patients at the beginning of the millennium]. AB - AIDS-related multidrug-resistant tuberculosis (MDRTB) emerged during the 90s in several countries around the world. In Argentina, the most notorious outbreak was documented in the Hospital Muniz, which is still undergoing its aftermaths. In order to evaluate the situation in this hospital regarding MDRTB, we analysed clinical, demographic and epidemiological traits of the 53 male MDRTB-aids patients admitted during 2001-2003 at a ward especially dedicated to their isolation. Patients' mean age was 32 years, 70% lived in Buenos Aires suburbs. A history of illicit drug users or imprisonment was recorded in 68% and 26% of the patients, respectively. Severe immunodepression (CD4+ count < 100/microl) was found in 88% of the patients and 58% died. Mortality was associated with non adherence to treatment and co-morbidity, but not with the genotype of the "M" strain, responsible for the original outbreak. Of 40 cases available for restriction fragment length polymorphism (RFLP), 29 (72.5%) resulted in cluster. RFLP patterns of 24 matched the "M" genotype. In this study, resistance to 5 or 6 drugs was found to be an indicator of disease due to the "M" strain. The "M" genotype associated significantly to previous admission at the Hospital Muniz or imprisonment. In brief, 14 years after the detection of the first MDRTB-aids case, we report here the persistence and predominance of the original outbreak strain at the hospital. Stronger TB infection control measures are urgently needed in hospitals and jails in order to strengthen the declining trend of the MDRTB observed in our country towards the end of the last decade. PMID- 17137169 TI - [Autoimmune-alloimmune neonatal neutropenia. Serum reactive IgG and neutrophil specific phenotype detected by flow cytometry]. AB - Auto or alloantibodies reactive with neutrophils define immune neutropenia. Alloimmune neonatal neutropenia is caused by maternal sensitization to paternal neutrophil antigens, resulting in IgG antibodies that are transferred to the fetus through the placenta. We present the studies in 4 children from 3 families with neutropenia of unknown origin (two of them were brothers). They were evaluated by flow cytometry in parallel with leukoagglutination. Reference values were established for serum reactive IgG in healthy volunteers for three dilutions (1/2, 1/5 and 1/20), both for the autologous reaction (serum and cells of the same individual) and for the heterologous reaction (serum and cells of different individuals). Results were expressed by an index defined by the quotient of the mean fluorescence intensity of the patient's serum divided by that of the reference serum. Serum reactive/agglutinant factors and circulating immune complexes were evaluated in patients and parents serum. Neutrophil specific phenotypes were determined for HNA-1a, HNA-1b and HNA-2a. Reactive IgG/agglutinant factors were found in 4 children. Two maternal sera were reactive against paternal and/or children neutrophils. Circulating immune complexes were detected in 2/4 children sera and were negative in 3/3 maternal sera. Maternal/children incompatibility was detected in the four cases. The three mothers had the same phenotype: homozygous NA1/NA1, NB1+. PMID- 17137170 TI - Ocular lesions and experimental choline deficiency. AB - Previous studies have shown ocular haemorrhages in choline-deficient rats. The aim of this paper is to study further the relationship between ocular and renal lesions and biochemical alterations in rats fed a choline-deficient diet. Fifty one weanling male Wistar rats, were divided into two groups. Thirty one of them were fed a choline-deficient diet and the rest was fed a choline-supplemented diet ad libitum. Animals from both groups were killed between the fifth and the eighth day. Urea, creatinine and homocysteine concentrations in blood were determined. Eyes were used for light microscopy study; high resolution light microscopy and the study of the retina as "retine a plat". Kidneys were studied by light microscopy. Choline-supplemented rats did not show ocular or renal lesion. Choline-deficient rats that showed renal lesions, tubular or cortical necrosis, did not always have ocular changes. There were no ocular changes in the only choline-deficient rat without renal lesion. The ocular changes consisted mainly in haemorrhage in both cameras and ciliary and vitreous bodies. Correlations between ocular and renal lesion (r = 0.72, p < 0.0001, CI 95%: 0.48 0.86); ocular lesion and creatinine (r = 0.86, p < 0.0001, Cl 95%: 0.72-0.93) and ocular lesion and urea (r = 0.70, p < 0.0001, Cl 95%: 0.44-0.85) were positive. Choline-deficiency induces ocular haemorrhagic lesions after the development of renal necrosis. The ocular pathology could be due to the immaturity of the ocular vasculature at this age. The hyaloid, choroid and retinal system are involved. PMID- 17137171 TI - [Mutations of hemochromatosis gene in volunteer blood donors and Chilean porphyria cutanea tarda patients]. AB - In patients with porphyria cutanea tarda (PCT), hepatic iron accumulation associated to hereditary hemochromatosis (HH) could play a role in the etiology and in the clinical expression of the disease. The H63D and C282Y mutations of the HFE gene frequency were studied in a PCT group of patients and compared with the frequency observed in a group of volunteer blood donors. PCT patients were cataloged as hereditary or acquired PCT carriers, whether or not they presented uroporphyrinogen decarboxilase gene mutations. Fifty percent of PCT patients were carriers of the disease's genetic type. Such percentage is significantly higher than what other authors have previously informed. H63D and C282Y mutations were present in 23% and 2.4% of the volunteer blood donors, respectively. Similar frequencies were informed by others authors in Chilean white ethnic populations, and also in Spaniard and Argentinean populations, but significantly higher than that observed in Chile's Araucanean aboriginal population. Probably the frequency of H63D and C283Y mutations are related to the Spaniard ascendancy dominance of Chile's white ethnic population. The frequency of HFE gene mutations in PCT patients was not different than what was observed in volunteer blood donors. Similarly, there was no statistical difference in the frequency of these mutations among patients with acquired or genetic PCT disease. With the obtained results, it is not possible postulate an association between PCT and the hereditary hemochromatosis of HFE gene mutations carrier conditions. PMID- 17137172 TI - [Surveillance of health care errors. An overview of the published data in Argentina]. AB - In the last decades, public health surveillance extended its scope of study to new fields, such as medical errors, in order to improve patient safety. This study was aimed to review all the evidence produced in Argentina about the surveillance of medical errors. An exhaustive literature search was performed. A total of 4656 abstracts were assessed (150 MEDLINE, 145 LILACS, and 4361 hand searched abstracts). Of them, 52 were analysed and 8 were considered relevant for health care error surveillance. Different approaches were used to study medical errors. Some of them have focused on patient safety and others on medical malpractice. There is still a need to improve the surveillance of this type of event. Mainly, the quality reports of study design and surveillance attributes were unclear. A critical appraisal and synthesis of all relevant studies on health care errors may help to understand not only the state of the art, but also to define research priorities. PMID- 17137173 TI - [Evaluation of diastolic function during and post-exercise in patients with arterial hypertension]. AB - It is known that patients with arterial hypertension and ventricular hypertrophy have diastolic alterations, in particular during exercise. However, it is controversial if diastolic dysfunction continues once exercise had concluded. The objective was to assess the effects of isometric exercise on the diastolic function in patients with arterial hypertension. Five control patients (group 1, G1) and 7 patients with arterial hypertension (group 2, G2) were studied. All patients underwent cardiac catheterization and performed isometric exercise until heart rate increased 43 +/- 7%. Left ventricular systolic pressure (LVSP) and end diastolic pressure (LVEDP) were measured. We calculated, +dP/dt(max), the time constant of isovolumic pressure decay (tau) and t1/2 were all measured. RESULTS: The LVSP increased in G1 and G2 during exercise from 140 +/- 3 to 195 +/- 14 mm Hg (p < 0.05) and 161 +/- 9 to 238 +/- 15 mm Hg, respectively; returning to their basal values once exercise had concluded. The tau (tau) and t1/2 increased, while exercising in G2, from 23 +/- 2 and 15 +/- 2 msec to 35 +/- 7 and 23 +/- 4 msec, respectively. After exercise both variables continued elevated reaching 41 +/- 6 msec (p < 0.05) and 23 +/- 3 msec (p < 0.05), respectively. In conclusion, isometric exercise decreases relaxation rate and increases LVEDP in patients with arterial hypertension and ventricular hypertrophy. After exercise, isovolumic relaxation remained altered suggesting the presence of stunned myocardium. PMID- 17137174 TI - [Paroxysmal sympathetic storm after diffuse axonal head injury]. AB - The term paroxysmal sympathetic storms is used to define episodic alterations in body temperature, blood pressure, heart and respiratory rate, size of pupil, and level of consciousness coinciding with hyperhidrosis, excessive salivation and extensor posturing. All the cases were presented after severe diffuse axonal head injury. We present two young patients with diffuse axonal head injury that develop in their evolution hypertension, tachycardia and fever without evidence during the episodes of epileptiform activity and without any infectious cause with excellent answer to the treatment with beta-blockers and morphine. We consider that the correct diagnosis of this entity minimizes the application of unnecessary studies allowing an appropriate treatment. PMID- 17137175 TI - [Community-acquired methicillin-resistant Staphylococcus aureus disseminated disease]. AB - A 21 year old man, previously healthy, presented with subcutaneous nodes consistent with gummas. Ultrasonography disclosed multiple subcutaneous abscesses and images suitable with piomiositis, pleural and pericardium effusion. A puncture-aspirate with fine-needle was performed and produced purulent material, with isolate of Staphylococcus aureus. Antimicrobial susceptibility testing by disk diffusion showed resistant to cefalotin, erythromycin and clindamycin, and susceptibility to trimethoprim-sulfamethoxazole, ciprofloxacin and rifampicin. Methicilin-resistance was confirmed by Staphyslide agglutination testing (Biomerieux). The patient was treated with ciprofloxacin and rifampicin during four weeks, with a good clinical response. The frequency of CA-MRSA infections is increasing, and these are reported in patients without identified predisposing risks leading to failure on empiric therapy for community infections presumed to be due to staphylococcal agents. PMID- 17137176 TI - [Spontaneous intracranial hypotension]. AB - Spontaneous intracranial hypotension is an infrequent clinical entity characterized by orthostatic headache, low cerebrospinal fluid pressure, and magnetic resonance imaging findings of diffuse pachymeningeal gadolinium enhancement without previous history of head trauma or lumbar puncture. A 24 year old healthy woman was referred after having daily headaches for two weeks. She described a headache that occurred soon after assuming an upright position and disappeared after resuming a recumbent position. The cerebrospinal fluid showed pleocytosis primarily lymphocitic, raised protein content and an opening pressure of 20 mm H2O. Magnetic resonance imaging of the head showed diffuse pachymeningeal gadolinium enhancement. Radioisotope cisternography using 99mTc DTPA confirmed the cerebrospinal fluid leak in the thoracic region. With conservative treatment the patient improved in a few days and the headache disappeared. Radiological findings, together with a compatible clinical condition, help to establish the diagnosis of spontaneous intracranial hypotension and avoid the use of unnecessary clinical investigations. PMID- 17137177 TI - [Two cases of enteritis with bacteremia due to Campylobacter jejuni]. AB - Campylobacter is an important agent of illness in human beings. Bacteremia occurs principally in the immunocompromissed host and is frequently due to C. fetus. Nevertheless bacteremia also has been observed in patients with enteritis due to C. jejuni. We refer two cases of patients with severe enteritis and bacteremia, both of them with immunosupressive concomitant diseases such as nephrotic syndrome and chronic cirrotic hepatopathy. Both patients presented hemathemesis. PMID- 17137178 TI - [Ischemic hepatitis]. AB - Ischemic hepatitis is an uncommon cardiovascular surgery complication. Hepatic biopsies show centrolobulillar necrosis. The term "hepatitis" was proposed because of a raise in hepatic enzymes similar with infectious disease, and "ischemic" because of failure in hepatic perfusion. Ischemic hepatitis was then defined as an acute and reversible elevation of hepatic enzymes (within 72 h), associated with disturbance in hepatic perfusion after excluding other causes of acute hepatitis. A 53 year-old male presented complaining of a 12 h epigastric pain, without nausea or vomiting, resistant to medication. He underwent an aortic valve replacement and was under anticoagulation. He suddenly developed shock and multiorgan failure. Jaundice and cardiac tamponade signs were present, associated with elevated hepatic enzymes. A transthoracic echocardiography accounted for cardiac tamponade signs. A pericardiocentesis was performed, removing 970 cc of hemorrhagic fluid, and hemodialysis, with improvement of his hemodynamic status. Hepatic enzymes improved. Viral markers were negative. PMID- 17137179 TI - [Ectopic mediastinal parathyroid adenoma. Detection with a radioisotopic probe and resolution with videothoracoscopy]. AB - We report a 66 years old woman with a diagnosis of primary hyperparathyroidism. Localization to mediastinum was obtained with parathyroid scintigraphy using 99mtc-methoxy-isobutyl-isonitrite (Tc99-MIBI). The patient was successfully operated upon by means of a videothora-coscopic approach. During the procedure serum parathormone was measured before and 10 minutes after adenomectomy, showing a more than 50% reduction from the basal level. An attempt to detect the precise site of the adenoma with a Tc99-MIBI probe was unsuccessful because of its proximity to the myocardium, but radioactivity was confirmed on the surgical specimen after resection. The patient's calcemia and parathormone levels became normal during the postoperative course and she remains normocalcemic 9 months after the procedure. In our case, preoperative localization and intraoperative parathormone measurements were both very useful for confirming surgical success; the intraoperative localization with a radioactive probe was not useful, but radioactivity was confirmed after resection on the surgical specimen. The endoscopic surgical procedure with videothoracoscopy was well tolerated, less painful than a thoracotomy, and it shortened the hospitalization period. PMID- 17137180 TI - [Malabsorption syndrome, growth disorder, mental retardation, premature aging and chronic renal insufficiency]. PMID- 17137181 TI - [Stroke in Fabry disease. More than a simple stenosis]. AB - The objective is to analyze the updated evidence on the physiopathological mechanisms that can generate cerebrovascular damage in Fabry disease. Fabry disease is the result of the deficiency of alpha-galactosidasa A, which causes pathological storage of glycosphingolipids, in different cells. Associated to renal and cardiac insufficiency, cerebrovascular complications can derive in the death of the patients. During a long time the only reported mechanism was the vascular occlusion by deposit of lipids at endothelial level. At the present time, other mechanisms are postulated. The arrival of enzyme replacement therapy has generated great expectation on the possibility of reversion of these alterations. Although the evidence is scarce and more long-term studies are necessary, some reports demonstrate that after months, the treatment has managed to revert some of the mechanisms involved. PMID- 17137182 TI - [Neuronal injury in multiple sclerosis]. AB - The concept of multiple sclerosis (MS) as a demyelinating disease is deeply ingrained. Although the existence of a neurodegenerative component has always been apparent, it has only recently become emphasized. Thus, in recent years several studies have identified axonal degeneration as the major determinant of irreversible neurological disability in patients with MS. Axonal injury begins at disease onset and remains clinically silent for many years; irreversible neurological disability develops when a threshold of axonal loss is reached and CNS compensatory mechanisms are exhausted. The precise mechanisms of axonal loss are poorly understood, and three hypotheses have been proposed: 1) The damage is caused by an inflammatory process, 2) There is an excessive accumulation of intra axonal Ca2+, 3) Demyelinated axons undergo degeneration due to lack of trophic support by myelin, or myelin forming cells. Although MS has traditionally been regarded as a disease of white matter, demyelination can also occur in the cerebral cortex. Cortical lesions exhibit neuronal injury represented by dendritic and axonal transection as well as neuronal apoptosis. Because conventional nuclear magnetic resonance (NMR) is limited in its ability to provide specific information about axonal pathology in MS, new techniques such as, diffusion-weighted MRI, proton magnetic resonance spectroscopy, functional MRI, as well as novel techniques designed to measure atrophy have been developed to monitor MS evolution. Recognition that MS is in part a neurodegenerative disease should trigger critical rethinking on the pathogenic mechanisms of this disease and provides new targets for a rational treatment. PMID- 17137183 TI - [Reflections on error in medicine]. PMID- 17137184 TI - [HeLa cells as prototype of immortalized cell lines]. PMID- 17137185 TI - [Global warming, ticks, and ehrlichiosis]. PMID- 17137186 TI - [Nobel Prize Physiology or Medicine. Acknowledging sharp clinical observation]. PMID- 17137187 TI - [Adjuvant effect of the Cry1Ac protoxin on cellular immunity when is coadministered with a good mucosal pathogen]. AB - BACKGROUND: The major part of pathogens use the mucosae to penetrate into the organisms, the idea to blockade the invasion, replication, colonization and multiplication of them results attractive not only to vaccines design, but immunopharmacs design, too. The protoxin Cry1Ac from Bacillus thuringiensis is a recent known mucosal adjuvant. OBJECTIVE: To determine the adjuvant effect of protoxin CrylAc from Bacillus thuringiensis on cellular response. MATERIAL AND METHODS: Mononuclear cells from human peripheral blood were isolated in Ficoll Hypaque gradients (d=1077), those cells CD3-CD14+ were obtained with the help of FACSVantage system, and then were stimulated with a lisate of S. pyogenes alone or combined with different concentrations of CrylAc. The proliferation of monocytes was determined in experiments of timidina-3H incorporation. RESULTS: The proliferative response of monocytes was major in those stimulated with S. pyogenes lisate and CrylAc than the observed when monocytes were stimulated with lisate alone. The major response was obtained with lisates of S. pyogenes plus 12.5 microg/mL of CrylAc (p<0.05). CONCLUSION: CrylAc have an important adjuvant effect on the cellular immune response. These findings are important to treat infections and for vaccines design. PMID- 17137188 TI - Meconium aspiration syndrome, parental atopy and asthma symptoms in children under two years old. AB - OBJECTIVE: To determine the association among meconium aspiration syndrome, parental atopy and asthma symptoms in children younger than two years. MATERIAL AND METHODS: One hundred thirty six children who had suffered meconium aspiration syndrome were followed from birth to they were two years old. CONTROL GROUP included 136 healthy children without meconium aspiration syndrome. RESULTS: Group of children with meconium aspiration syndrome: The average hospital stay in neonatal intensive care unit was of 5.91 +/- 4.44 days and were given 65.5 +/- 16.6 hours of oxygen. Prevalence of asthma symptoms during the last year was of 41.2%; 35.7% had family history of allergies (p = NS). The average age at the beginning of asthma symptoms was of 12.64 +/- 6.96 months; 78.6% of children were male. CONTROL GROUP: 30.9% had wheezing episodes during the last 12 months; 28.6% were male; 38.1% had family history of allergies (p = NS). The average age at the beginning of asthma symptoms was of 15.57 +/- 6.05 months. CONCLUSION: Aspiration of meconium seems to be an important risk factor of the early beginning of asthma symptoms in children younger than two years. Data shows that the prevalence of asthma symptoms was higher in case group than in control group. Asthma symptoms risk increases in children with aspiration meconium syndrome and family background of allergies. PMID- 17137189 TI - Asthma in emergency department. Guidelines, physicians and patients. AB - BACKGROUND: Morbidity and mortality by bronchial asthma continues to be a serious public health problem all over the world. Bronchial asthma is considered the most common chronic disease among children and asthmatic crises are the most frequent cause for visits to the emergency room. Among adults, bronchial asthma has also a high rate of morbidity and repercussions in productivity as well as in the costs of the health systems that assist them. On the other hand, despite the development of the International Guidelines for the DIAGNOSIS and Treatment of Bronchial Asthma (IGDTBA), supported by the best scientific medical researches based on evidence, which recommend the prompt and regular use of inhaled corticosteroids in the treatment of the persistent clinical forms of bronchial asthma to reduce their morbidity and mortality, this has not yet been modified. OBJECTIVE: To determine the causes of high morbidity and mortality due to bronchial asthma. MATERIAL AND METHOD: We retrospectively studied a group of 152 asthmatic patients (a pediatric group of 88 patients and a adult group with 64) who visited NOVA Clinic in Monterrey, Mexico, emergency medical service 234 times during one year (January 1st to December 31st, 2004). Four variables were investigated: a) Number of patients who were diagnosed with bronchial asthma following the present guidelines (IGDTAB). b) Number of patients prescribed regular treatment with inhaled corticosteroids. c) Number of patients who had good acceptance of regular treatment with inhaled corticosteroids (compliance). d) The existence of some type of communication or cooperation among the different levels of medical attention (emergency service, outpatient service and asthma specialists) allowing them to work as a team in the treatment and education of the asthmatic patient. RESULTS: a) DIAGNOSIS: only one patient of the pediatric group and seven of the adult group had a diagnosis based on the IGDTBA. b) Only 42.7% of the patients were prescribed inhaled corticosteroids by their physicians as a regular preventive anti-inflammatory treatment. c) Only 47.6% of the patients who were prescribed inhaled corticosteroids as a regular treatment followed the same. When adding together the last two variables, it is observed that only 23 out of 88 patients in the pediatric group (26%) and 8 out of 64 adults (12.5%) followed regular treatment with inhaled corticosteroids as is recommended in the guidelines (IGDTAB). d) We found no communication or cooperation among the different levels of medical attention for the education and treatment of any of the patients studied. CONCLUSIONS: The persistent morbidity from bronchial asthma in our study is due to: a) Most of physicians who care for asthmatic patients in emergency or outpatient services are not familiar with the International Guidelines for the DIAGNOSIS and Treatment of Bronchial Asthma (IGDTAB) and those who state that they do know about them have no interest in using them with their patients. b) Inhaled corticosteroids, recommended as the first line of anti-inflammatory medications for regular treatment of bronchial asthma, are not used by the majority of physicians. c) Asthmatic patients who follow regular treatment with inhaled corticosteroids account for lesser than 50%. d) There is no communication or cooperation among the different levels of medical attention for asthmatic patients (emergency medical service, outpatient service and specialists in asthma) for their education and treatment under the same parameters (IGDTBA), which partially explains the persistent morbidity and visit to emergency department. PMID- 17137190 TI - [Allergic fungi: importance of the standardization of fungal extracts and their application on clinical practice]. AB - Among the aeroallergens associated to asthma and allergic rhinitis, the fungi are a common cause of diagnostic and therapeutic problems. The wide variety and distribution of fungal species and the complex characterization of their allergenicity, is a complex item. The fungus extracts used to diagnose and treat sensitizations are frequently non effective, and different varieties of extracts are globally distributed. The standardization of commercial fungi extracts results extremely important as diagnostic procedure as well as to decide an efficacious and safe immunotherapy. This paper reviews important methodological steps to the standardization of fungi extracts, and finally the clinical use of these extracts. PMID- 17137191 TI - [When the cause of dyspnea is on larynx. Asthma of difficult control, resistant to treatment? Vocal cords dysfunction? or Both?]. AB - Dyspnea is a symptom whose diagnosis requires the exclusion of other diseases with which it can be confused, such as asthma and a variety of pulmonary and heart diseases. Dyspnea originated in the larynx may occur due to obstruction by a tumor or other affections in situ, interfering the airway, such as: edema by infections or inflammatory processes, a traumatism, an abnormal movement of the larynx structures, such as the inappropriate or absent of the vocal cords or the laryngospasm. Severity of larynx dyspnea may be to mild to acute, risking the life. This paper reviews the normal laryngeal function and the anatomic, obstructive, and functional disorders that can lead to dyspnea. Some suggestions are also made to determine the cause and treat these diseases. PMID- 17137192 TI - [Study on factors related to suicide--from the viewpoint of psychiatry. Suicide by the aged and children. Nos. 1 and 2]. PMID- 17137193 TI - [Discharge curve among psychiatric patients after admission and risk factors associated with long stay based on "patient survey"]. AB - The "Reform Vision of Mental Health Services" (2004) announced the basic policy for the transition from hospital based to community based care, and set up numerical objectives, such as the average proportion remaining hospitalized in the first year after admission and the incidence rate of discharge among psychiatric patients hospitalized for more than one year. Using data from the "Patient Survey" performed in 2002 by the Ministry of Health, Labour and Welfare, we estimated discharge curves for each mental disorder during the first year after admission and assessed the effects of variables, i.e., diagnosis, sex, age, hospital type, and residential area, on remaining hospitalized after one year from admission and the incidence rate of discharge among psychiatric patients hospitalized for more than one year. The estimated number of discharged psychiatric patients was 27,974 in September, 2002, and 86% of them were discharged less than one year after admission. The incidence rate of discharge (per 100 person-year) in the first year was 314.8, but the rate after the second year sharply decreased to 19.9. Patients with dementia, mental retardation, and schizophrenia tended to stay for a long period in hospital, and proportions remaining hospitalized after one year from admission were 27.0%, 16.4%, and 14.6% respectively. Based on multivariate analysis using the weighted Poisson regression model, risk factors associated with an increased chance of remaining hospitalized after the first year included a long length of continuous hospitalization, diagnoses of dementia, mental retardation, and schizophrenia, male, older age, and being in a mental hospital. On the other hand, as to the incidence rate of discharge after one year, a long length of continuous hospitalization and being in a mental hospital were related with a long stay, but other variables were slightly different. Being female, patients aged 45-54 years old, and diagnoses of epilepsy and schizophrenia were associated with a long stay. These results clarify the present situation of discharge among psychiatric inpatients and indicate the important variables associated with discharge to prevent new long hospital-stay cases in Japan. PMID- 17137194 TI - [A clinical psychiatric study about post-stroke depression]. AB - We examined the clinical characteristics of post stroke depression (PSD) patients in a rehabilitation hospital. The subjects were inpatients at the Ibaraki Prefectural University of Health Sciences Hospital who had been diagnosed with stroke and admitted between May 2002 and January 2004. Of 123 patients enrolled, 18 were diagnosed with PSD. We investigated the clinical differences between PSD patients and non-depressed (ND) patients. There were no significant differences between PSD and ND groups in terms of sex, age, past psychiatric history, family psychiatric history, stroke diagnosis, or neurological symptoms. The scores of PSD patients on Zung's self-rating depression scale and the Hamilton depression rating scale were significantly higher than those of ND patients. Furthermore, the Activities of Daily Living measured by the Functional Independence Measure and cognitive function evaluated by the Mini-Mental State Examination of ND patients were significantly better than those of PSD subjects. These results suggest that the detection of PSD in stroke patients is important for promoting their rehabilitation and improving their Activities of Daily Living and cognitive function. PMID- 17137195 TI - [Training in clinical psychiatry--from the viewpoint of the Sub-Committee on Postgraduate Education, Committee on Psychiatric Specialities]. PMID- 17137196 TI - [For the preparation for psychiatric specialist training programs: from a viewpoint of university hospitals]. PMID- 17137197 TI - [Specific methods and problems related to clinical training in psychiatry--the attitude of private psychiatric hospitals]. PMID- 17137198 TI - [Life-time education for psychiatric specialists]. PMID- 17137199 TI - [Life-time education of psychiatric specialists--from the viewpoint of university hospitals]. PMID- 17137200 TI - [Life-long education of a specialists is possible only through daily activities as clinical instructors being evaluated by interns and mutual efforts for advancement]. PMID- 17137201 TI - [Possibility for regional psychiatric care in Tohoku and Hokkaido areas]. PMID- 17137202 TI - [Occupations and geographic areas: psychiatric care in the Tohoku area conducted by the National Defense Forces]. PMID- 17137203 TI - [The current status and objectives for psychiatric medicine in Chugoku and Shikoku areas: a view from affiliated medical organizations]. PMID- 17137204 TI - [The current status and objectives of the acute care wards of university hospitals (from the view of psychiatrists in Iwate Prefecture)]. PMID- 17137205 TI - [The current status of psychiatric care in 4 prefectures in the southern Kanto Area and problems specific to the area]. PMID- 17137206 TI - [Interpretation and the use of evidence in psychiatry]. PMID- 17137207 TI - [Treatment of schizophrenia with emphasis on adherence--for the prevention of recurrences]. PMID- 17137208 TI - [Symptomatic psychoses]. PMID- 17137209 TI - Control of morning blood pressure: the best preventive strategy against stroke. PMID- 17137210 TI - Impairment of instantaneous autonomic regulation relates to blood pressure fall immediately after standing in the elderly and hypertensives. AB - The relation between changes in blood pressure and changes in autonomic activity over a very short period of time has not been reported thus far. To examine this relation, we here introduced a new method of power spectrum analysis with wavelet transformation, which has very fine time resolution and is able to assess changes in autonomic activity quantitatively even during movement. Our subjects were 15 hypertensive and 17 normotensive subjects. A head-up tilt test was performed in all subjects, and during the test, electrocardiogram and blood pressure were recorded continuously. The power spectrums for both parameters were calculated simultaneously every 5 s using wavelet transformation. The high frequency of the RR interval of the electrocardiogram (RR-HF) and low frequency of systolic blood pressure (SBP-LF) were defined and calculated as markers of parasympathetic and alpha-1 receptor blocker, bunazosin-sensitive sympathetic activity, respectively. Focusing on the changes for 2 min immediately after head-up tilting, it was found that the changes in SBP-LF and RR-HF were significantly delayed, by at least 40 s, in hypertensives compared with normotensives and also in elderly compared with non-elderly subjects. Multiple regression analysis demonstrated that the instantaneous change in RR-HF was the most important confounding factor for a fall in blood pressure immediately after head-up tilting. In conclusion, real time changes in autonomic activity calculated by wavelet transformation may provide sensitive and useful information about acute changes in cardiovascular regulation, such as delayed reaction of the autonomic regulation after head-up tilting, that may be major causes of the blood pressure fall in hypertensive and elderly subjects. PMID- 17137211 TI - Renal and vascular protective effects of telmisartan in patients with essential hypertension. AB - It is known that the angiotensin receptor blockers (ARBs) have organ protective effects in patients with heart failure or renal impairment. Several studies have revealed that the ARB telmisartan has an organ protective effect, but there have been few studies directly comparing the effects of telmisartan and calcium antagonists, since most clinical studies on telmisartan have been conducted in treated patients or patients on combination therapy. The present study was conducted to compare the renal and vascular protective effects of telmisartan monotherapy and calcium antagonist monotherapy in untreated hypertensive patients. Forty-three patients with untreated essential hypertension were randomized to receive amlodipine (n=22) or telmisartan (n=21), which were respectively administered at doses of 5 mg and 40 mg once daily in the morning for 24 weeks. The patients were examined before and after treatment to assess changes of renal function, flow-mediated dilation (a parameter of vascular endothelial function), and brachial-ankle pulse wave velocity (baPWV; a parameter of arteriosclerosis). Before treatment, there were no significant differences in these parameters between groups. The decreases of urinary albumin excretion and baPWV, and the increase of flow-mediated dilation were significantly greater in the telmisartan group than the amlodipine group, while the antihypertensive effects were not significantly different between the two groups. In conclusion, these results suggest that telmisartan is more effective at protecting renal function and vascular endothelial function, and at improving arteriosclerosis than the calcium channel blocker in patients with essential hypertension. PMID- 17137212 TI - Synthetic/secreting and apoptotic phenotypes in renal biopsy tissues from hypertensive nephrosclerosis patients. AB - The major glomerular abnormalities in hypertensive nephrosclerosis are described as glomerular obsolescence (GO), glomerulosclerosis (GS), and glomerular collapse (GC). However, glomerular cellular changes caused by hypertensive insults have not been well analyzed. Using an immunoenzyme method, we examined eleven biopsy samples from patients with hypertensive nephrosclerosis for two synthetic and secreting phenotypes, a-smooth muscle actin (alpha-SMA) and collagen type III (Col. III), and two apoptotic phenotypes, pro-apoptotic molecule Bax and anti apoptotic molecule BcI-2. Together with the glomerular and vascular changes and interstitial fibrosis (IF) area, the results were scored quantitatively and semi quantitatively and compared to the clinical findings, which included systolic blood pressure (SBP), mean arterial pressure (MAP), serum creatinine levels (sCr) and creatinine clearance (Ccr), using univariate and multivariate analyses. As a result, GS was frequently observed in the mild-to-moderate hypertensive group (140 < or = SBP<180 mmHg), whereas GC was positively correlated with SBP. Furthermore, there was a positive correlation of GS with mesangial alpha-SMA and Col. III, suggesting that GS was the reflection of these synthetic and secreting phenotypic changes in mesangial cells. Endothelial Bax was positively correlated with Ccr (p<0.01); in contrast, podocytic Bax was positively correlated with sCr (p<0.05) and showed a tendency to correlate with MAP (p=0.054). In conclusion, these findings support the view that mesangial synthetic and secreting phenotypic changes may be a reflection of cellular activation caused by mild-to-moderate hypertension and that apoptotic phenotypic expression in podocytes, rather than endothelial cells, may be related to the development of a severe form of hypertensive nephrosclerosis. PMID- 17137213 TI - Morning hypertension: the strongest independent risk factor for stroke in elderly hypertensive patients. AB - Stroke occurs most frequently in the morning hours, but the impact of the morning blood pressure (BP) level on stroke risk has not been fully investigated in hypertensives. We studied stroke prognosis in 519 older hypertensives in whom ambulatory BP monitoring was performed, and who were followed prospectively. During an average duration of 41 months (range: 1-68 months), 44 stroke events occurred. The morning systolic BP (SBP) was the strongest independent predictor for stroke events among clinic, 24-h, awake, sleep, evening, and pre-awake BPs, with a 10 mmHg increase in morning SBP corresponding to a relative risk (RR) of 1.44 (p<0.0001). The average of the morning and evening SBP (Av-ME-SBP; 10 mmHg increase: RR=1.41, p=0.0001), and the difference between the morning and evening SBP (Di-ME-SBP; 10 mmHg increase: RR=1.24, p=0.0025) were associated with stroke risks independently of each other. The RR of morning hypertension (Av-ME-SBP > or = 135 mmHg and Di-ME-SBP > or = 20 mmHg) vs. sustained hypertension (Av-ME-SBP > = 135 mmHg and Di-ME-SBP < or = 20 mmHg) for stoke events was 3.1 after controlling for other risk factors (p=0.01). In conclusion, morning hypertension is the strongest independent predictor for future clinical stroke events in elderly hypertensive patients, and morning and evening BPs should be monitored in the home as a first step in the treatment of hypertensive patients. PMID- 17137214 TI - Metabolic syndrome, C-reactive protein and increased arterial stiffness in Japanese subjects. AB - The aim of this study was to investigate whether the metabolic syndrome (MS) was associated with an elevated level of C-reactive protein (CRP) and increased arterial stiffness, and to clarify whether combined MS and CRP data had a stronger relation to arterial stiffness than did MS data alone. Brachial-ankle pulse wave velocity (baPWV), CRP, and conventional risk factors were evaluated in 3,412 men and 854 women. Adjusted mean values of baPWV in men with 0, 1, 2, and > or = 3 components were 1,309, 1,372, 1,422, and 1,462 cm/s, respectively (p for trend <0.001). Adjusted mean values of baPWV in women with 0, 1, 2, and > or =3 components were 1,212, 1,292, 1,357, and 1,391 cm/s, respectively (p for trend <0.001). Adjusted geometric mean concentrations of CRP in men with 0, 1, 2, and > or = 3 components were 0.036, 0.049, 0.059, and 0.076 mg/dI, respectively (p for trend <0.001). Adjusted geometric mean concentrations of CRP in women with 0, 1, 2, and > or = 3 components were 0.023, 0.030, 0.057, and 0.077 mg/dI, respectively (p for trend <0.001). In analyses of adjusted mean values of baPWV according to the number of MS components and according to CRP levels within or without top quartile levels, the p value for the trend was significant (<0.001) in both men and women but, in post hoc analyses, comparing high and low CRP levels in each MS component-number group, no significant difference was found. These results suggest that, for prediction of increased arterial stiffness, combining MS and CRP data has little additive effect compared to the use of MS data alone. PMID- 17137215 TI - Plasma aldosterone in hypertensive patients on chronic hemodialysis: distribution, determinants and impact on survival. AB - A high plasma aldosterone concentration (PAC) is known to be associated with poor outcome in patients with cardiac disease. However, the prognostic value of PAC in chronic hemodialysis (HD) patients is unknown. In 1996 we examined 128 hypertensive patients treated with antihypertensive drugs, excluding angiotensin converting enzyme inhibitors, who were undergoing chronic HD (ages 61.8+/-13.8 years, 62% male), and for whom PAC (ng/dl) data were obtained. We followed up these patients until November 2003. During the follow-up period, 30 patients died. About half of all patients (48%) had PAC values above the normal range. We assigned the 128 patients to a lower (<22.9) or higher (> or = 22.9) PAC group according to the median baseline PAC. The survival rate as calculated by the Kaplan-Meier method was 90.6% in the higher PAC group and 62.5% in the lower PAC group (p=0.003). In multivariate analysis, serum potassium and plasma renin activity were independent determinants of PAC. Cox proportional hazards analysis, with adjustment for other variables including diabetes, showed that lower PAC was independently predictive of death. The adjusted hazard ratio (95% confidence interval) of the lower PAC group was 2.905 (1.187-7.112, p=0.020). The significance of PAC became marginal by adjustment with albumin or potassium. These results indicate that higher PAC is common, but not associated with an increase in total and cardiovascular deaths among hypertensive patients undergoing chronic HD. The association between lower PAC and poor survival may be driven by volume retention and/or lower potassium. PMID- 17137216 TI - Relationships between sleep-disordered breathing and blood pressure and excessive daytime sleepiness among truck drivers. AB - Sleep-disordered breathing is a risk factor for hypertension, cardiovascular disease and accidents in the general population, but little is known about this correlation among professional truck drivers. To examine the relationships of sleep-disordered breathing with blood pressure levels and excessive daytime sleepiness among truck drivers, we conducted a population-based cross-sectional study of 1,313 subjects aged 20-69 years registered in the Japanese Trucking Association. The 3% oxygen desaturation index was selected as an indicator of sleep-disordered breathing, representing the number of desaturation events per hour of recording time in which blood oxygen fell by > or = 3% by overnight pulse oximetry. The Epworth Sleepiness Scale was used to estimate excessive daytime sleepiness. There were significant positive associations between the 3% oxygen desaturation index levels and both diastolic blood pressure levels and Epworth Sleepiness Scale scores. The multivariate odds ratio of hypertension was 2.0 (1.1 3.6) for a 3% oxygen desaturation index of > or = 15 in reference with a 3% oxygen desaturation index of <5. This association was more evident among those aged > or = 40 years and overweight subjects. Further, the multivariate odds ratio of an Epworth Sleepiness Scale of > or = 11 was 2.3 (1.1-4.9) for a 3% oxygen desaturation index of > or =15 in reference with a 3% oxygen desaturation index of <5. This association was more evident among those aged > or =40 years. The associations of sleep-disordered breathing severity with diastolic blood pressure levels and excessive daytime sleepiness suggest the need for sleep disordered breathing screening among truck drivers for prevention of hypertension and potential traffic accidents. PMID- 17137217 TI - Association of sixty-one non-synonymous polymorphisms in forty-one hypertension candidate genes with blood pressure variation and hypertension. AB - We previously selected a group of hypertension candidate genes by a key word search using the OMIM database of NCBI and validated 525 coding single nucleotide polymorphisms (SNPs) in 179 hypertension candidate genes by DNA sequencing in a Japanese population. In the present study, we examined the association between 61 non-synonymous SNPs and blood pressure variations and hypertension. We used DNA samples taken from 1,880 subjects in the Suita study, a population-based study using randomly selected subjects. Analyses of covariance adjusting for age, body mass index, hyperlipidemia, diabetes, smoking, drinking, and antihypertensive medication revealed that 17 polymorphisms in 16 genes (APOB, CAST, CLCNKB, CTNS, GHR, GYS1, HF1, IKBKAP, KCNJ11, LIPC, LPL, P2RY2, PON2, SLC4A1, TRH, VWF) were significantly associated with blood pressure variations. Multivariate logistic regression analysis with adjustment for the same factors revealed that 11 polymorphisms in 11 genes (CAST, CTLA4, F5, GC, GHR, LIPC, PLA2G7, SLC4A1, SLCI8A1, TRH, VWF) showed significant associations with hypertension. Five polymorphisms in five genes, CAST(calpastatin), LIPC (hepatic lipase), SLC4A1 (band 3 anion transporter), TRH (thyrotropin-releasing hormone), and VWF (von Willebrand factor), were significantly associated with both blood pressure variation and hypertension. Thus, our study suggests that these five genes were susceptibility genes for essential hypertension in this Japanese population. PMID- 17137218 TI - Valsartan improves the lower limit of cerebral autoregulation in rats. AB - The effects of angiotensin II type 1 receptor blockers (ARBs) on cerebral blood flow (CBF) autoregulation have not been fully clarified. Thus, we examined the acute effect of valsartan, the most selective ARB, on CBF autoregulation in spontaneously hypertensive rats. Intravenous administration of valsartan (0.3 mg/kg) reduced the mean arterial pressure (MAP) from 184+/-5 (mean+/-SEM) to 174+/-5 mmHg (p<0.001) without affecting CBF as measured by laser-Doppler flowmetry. The lower limit of CBF autoregulation (the MAP at which the CBF was 80% of the baseline value) in the valsartan-treated group (122+/-3 mmHg) was significantly lower than that in the control group (135+/-4 mmHg, p<0.05). Reverse transcribed-polymerase chain reaction and immunohistochemical staining demonstrated that both angiotensin II type 2 receptors and angiotensin II type 1 receptors (AT1Rs) were expressed in endothelial and smooth muscle cells of the rat cerebral arteries. These results suggest that specific inhibition of AT1Rs in the cerebral circulation causes the leftward shift of the lower limit of autoregulation. PMID- 17137219 TI - Effects of eplerenone and salt intake on left ventricular remodeling after myocardial infarction in rats. AB - Eplerenone, a selective aldosterone blocker, has been shown to attenuate cardiac fibrosis and decrease cardiovascular events in both experimental and clinical studies. We examined the cardioprotective effect of eplerenone in myocardial infarction (MI) rats receiving different levels of salt in their diet. The MI rats were randomly divided into five groups: Group CL, animals received a low salt diet (0.015%); Group EpL, a low-salt diet with eplerenone (100 mg/kg/day in food); Group CH, a high-salt diet (0.9%); Group EpH, a high-salt diet with eplerenone; and Group C, a normal salt diet (0.3%). These diets were continued for 4 weeks. Echocardiographic and histomorphological examinations revealed that the administration of eplerenone significantly improved the cardiac function, significantly suppressed compensatory cardiac hypertrophy and significantly reduced cardiac fibrosis in both the interstitial and the perivascular areas in the high-salt diet group (Group EpH). However, eplerenone had no observable effects in the low-salt diet group (Group EpL). Also, these examinations demonstrated that the left ventricular remodeling after MI was suppressed and the cardiac function was improved in the group receiving a low-salt diet without eplerenone (Group CL), even though there was a significant increase of aldosterone level in blood, in comparison to the group receiving a high-salt diet without eplerenone (Group CH). These results indicate that the cardioprotective effect of eplerenone varies depending on the salt intake. PMID- 17137220 TI - [Unsuccessful laparoscopic castration in a cryptorchid Frisian stallion]. AB - A 2-year-old unilateral inguinal cryptorchid Frisian stallion was castrated by bilateral laparoscopic intra-abdominal spermatic cord transection. Because blood testosterone levels were still high on postoperative day 7, both testicles were removed using an open approach. Pathology revealed the presence of vital Leydig cells in both testes. Failure of the laparoscopic castration in this horse was attributed to an alternate blood supply to the testes by vessels derived from the cremaster and/or external pudendal artery. PMID- 17137221 TI - [O2K]. PMID- 17137222 TI - [Analysis of the article 'Homeopathy, a solution to diarrhea in calves!']. PMID- 17137223 TI - [Official start of Petplan on world animal day]. PMID- 17137224 TI - [Exclusion of classical swine fever by PCR test]. PMID- 17137225 TI - [Brussels makes decision about the criteria of non-POM (prescription-only medicines) and drugs for horses]. PMID- 17137226 TI - [Vaccination... Health by injection?]. PMID- 17137227 TI - [Speech 2006]. PMID- 17137228 TI - [Professional integrity proves to be a challenging theme]. PMID- 17137229 TI - Case managers need extensive education, regular competency exams. PMID- 17137231 TI - Tracking delays cuts LOS, improves patient flow. PMID- 17137230 TI - Education helps CMs learn to do their jobs better. PMID- 17137232 TI - Demonstration project claims dollar 1 billion in potential savings. PMID- 17137233 TI - Project participants reaping the benefits. PMID- 17137234 TI - Team educates staff, ensures compliance. PMID- 17137235 TI - Don't skimp on evaluation. PMID- 17137236 TI - Elderly AMI patients don't always get beta-blockers. PMID- 17137237 TI - The economic effect of switching from sildenafil to another phosphodiesterase type 5 inhibitor. AB - Patients who could benefit from additional education about treatments for erectile dysfunction (ED) may prematurely discontinue or switch ED medications, resulting in unnecessary resource utilization. In a retrospective cohort study using a large, aggregated health claims database, the costs associated with switching from sildenafil to another phosphodiesterase type 5 (PDE-5) inhibitor were compared with those for patients refilling sildenafil. Of the 15,584 patients with an index sildenafil claim, 10,863 had a second PDE-5 inhibitor prescription (10,137 for sildenafil, 726 for vardenafil or tadalafil). Erectile dysfunction-attributable costs in the six-month preindex period were similar (P = .72), but postindex six-month ED costs were higher in patients who initially switched from sildenafil (dollar 173.38) versus patients who refilled sildenafil (dollar 131.51; P < .001). Regression analysis estimated that corrected ED attributable and overall costs were 41% (P < .001) and 43% (P < .001) higher for patients who switched versus those who refilled sildenafil, respectively. PMID- 17137238 TI - The effect of formularies on prescribing beyond the health plan. PMID- 17137239 TI - Spillover effects of restrictive drug formularies in the statin class: a descriptive study of the Medicaid preferred drug list in Texas. AB - This study examines whether the implementation of Texas' Medicaid preferred drug list (PDL) influenced, or had a spillover effect, on the prescribing of statin products for non-Medicaid patients. Simvastatin and pravastatin were on the PDL, but atorvastatin was not. Physicians in Texas were categorized into four groups with increasing Medicaid exposure. Compared with physicians with very low, low, or medium Medicaid exposure, the non-Medicaid marketshare of atorvastatin dropped approximately two percentage points and the non-Medicaid marketshare of simvastatin and pravastatin increased by approximately one percentage point among physicians with high Medicaid exposure after six months. Therefore, the restrictive Medicaid PDL in Texas spilled over to non-Medicaid patients. PMID- 17137240 TI - Medical tourism: a role disease management? PMID- 17137241 TI - The effect of recombinant activated factor VII in the treatment of intracerebral hemorrhage on health plan budgets. AB - Treating patients with intracerebral hemorrhage (ICH) using recombinant activated factor VII (rFVIla) has been found to improve survival and functional outcome. To examine how the introduction of rFVIla 80 microg/kg as a treatment for ICH affects the budget of a health plan, a decision-analysis model was developed which considered both short-term hospitalization costs and long-term management of disability. Assuming a health plan enrollment of 1 million members and initial rFVIla uptake of 50% in appropriate patients, the annual health plan cost may be expected to increase by dollar 64,781 (dollar 0.005 per-member per-month). With a 5% increase in uptake each year, the annual health plan's cost may decrease compared with the current budget within three years. The implications for this sample health plan's budget are modest in the first year, and a reduction in costs is expected within three years owing to improved functional outcomes of patients. PMID- 17137242 TI - Formulary implications of management of pulmonary arterial hypertension: Part I- An overview of existing and new pharmacological treatment options. AB - Pulmonary arterial hypertension (PAH) represents a significant challenge to physicians, particularly because of its etiology and nonspecific presentation. Unfortunately, the majority of patients are not diagnosed until the condition is at an advanced stage. In this two-part White Paper from The Pharmacy and Therapeutics Society, prostanoids, administered as continuous intravenous or subcutaneous infusions, are the most effective treatments but are inconvenient and costly, owing to complex parenteral administration. The endothelin-receptor antagonist bosentan and the phosphodiesterase-5 inhibitor sildenafil are currently the only oral therapies licensed for PAH; however, bosentan has an identified risk of liver toxicity. Newer PAH treatments may offer benefits over existing therapies in terms of less potential for liver damage and more favorable interaction profiles. PMID- 17137243 TI - Is managed care education for new nurses adequate? AB - Nursing practices have been affected by managed care at all levels, from nursing administration to direct patient care, causing one to ask if new practicing nurses receive adequate managed care education. A search of the literature published between 1982 and February 2006 was conducted using ProQuest, Article First, ERIC, and PubMed. The literature primarily comprised anecdotal articles; only two qualitative research-based studies were published. The search revealed nurse employers and nurse educators believe that managed care competencies are very important for new practicing nurses, but even with curricula changes, managed care competencies are deficient for new practicing nurses. Further research is recommended to identify the specific managed health care education that would be effective in preparing new practicing nurses for today's health care arena at all levels of nursing. PMID- 17137244 TI - [Will therapeutic cloning soon become legal in France?]. PMID- 17137245 TI - [Janeway erythremic hand lesions]. PMID- 17137246 TI - [Head and neck cancer: towards a more effective and less damaging therapeutic strategy]. PMID- 17137247 TI - [Epidemiology of head and neck cancer]. AB - In 2000, 19,600 new cases of head and neck carcinomas, oral cavity, pharynx and larynx cancer occurred in the French population. In men, these tumours are predominant and represent the 4th cancer incidence rank with 16,855 new cases, and the 5th cancer mortality rank with 6309 deaths. For women, in 2000, the incidence was 2830 new cases with 900 deaths (14th rank among women cancer). The decrease of alcohol and tobacco consumption and the improvement of treatment efficacy explain the decreasing head and neck cancer incidence and mortality in the past decades in France. PMID- 17137248 TI - [Presentation and diagnostic evaluation of head and neck cancer]. AB - Dysphagia, dysphonia, dyspnoea, cervical lymph nodes are commonly indicative of head and neck carcinoma, although non specific signs, like otologic or rhinologic symptoms may be present. In addition to physical examination done on the patient awake and panendoscopy under anesthesia with biopsies, the diagnostic evaluation includes different imaging modalities (computed tomography, and/or magnetic resonance imaging, and positron emission tomography with FDG). Preoperative assessment of tumour extent, cervical lymph node involvement as well as distant metastases is mandatory for both staging and planning the best accurate therapy in these patients. PMID- 17137249 TI - [Current trends in diagnosis and treatment head and neck cancer]. AB - Prognosis of head and neck squamous cell carcinomas (HNSCC) is partly determined by lymph nodes metastases. Contemporary imaging provides detection of adenopathies but remains unable to detect infracentimetric micrometastatic nodes. Topographical classification according to Robbins allows for better treatment through proper definition of impaired neck areas. The high rate of node metastases in HNSCC indicates systematic treatment of the neck in most localizations. Several neck dissection techniques are available with wide acceptation of elective functional neck dissections. Improvement of detection of node metastases and better selection of neck sites for treatment are eventual direction of progress. In neck carcinoma with unknown primary, search of primary should address first head and neck area and upper aero-digestive tract. Differential diagnosis are glomic tumour, congenital cyst and other causes of lymph node disease PMID- 17137250 TI - [Molecular markers of head and neck cancer]. AB - Molecular markers can be used for the diagnostic (early detection, assessment of tumor extent, detection of residual disease), for the prognostic (disease classification and therapeutic indications), for the treatment (treatment target and predictive factors of tumor response to treatment) and for the prevention (identification of the at-risk population and chemoprevention). Molecular markers have to be validated by prospective studies using appropriate methods. Currently, no molecular marker is used in routine practice for the management of upper aerodigestive tract cancers, but new techniques are being developed to analyze simultaneously tens of thousands of markers; they should expand our knowledge of the biology of these cancers. PMID- 17137251 TI - [Principal treatments of head and neck cancer]. PMID- 17137252 TI - [Organ preservation surgery in pharyngolaryngeal cancer]. AB - Organ preservation surgery is a reasonable name for these types of techniques because it accurately describes the postoperative outcome, which is preservation of the quintessential laryngeal functions: swallowing, respiration without a permanent stoma and speech. In response to the common use of total laryngectomy, the non-surgical approaches have often been referred to as organ preservation strategies. In many institutions, it appears that there are organ preservation strategies and then there is surgery. In our opinion there are both non surgical organ preservation strategies and surgical organ preservation strategies. The key is that in both approaches the goal is to spare the functions of the larynx. In both, the work-ups are very important to analyse 3-dimensional extent of the lesion by nasofibroscopy (laryngeal mobilities), endoscopy and radiology. Organ preservation surgical approach allowed to save the use of radiotherapy for the management of various metachronous second primary tumours. PMID- 17137253 TI - [Non surgical treatment approach for head and neck cancer]. AB - In many patients with localized carcinoma, radiotherapy alone is an alternative to surgery. In patients with non surgical locally advanced carcinoma of the head and neck the use of concurrent chemotherapy and radiotherapy is a standard. Chemotherapy is a platinum-based chemotherapy. Radiotherapy must be delivered using 3D conformal technique with or without intensity modulation to increase local control and/or overall survival. An alternative for chemotherapy is an induction use for organ preservation. If chemotherapy is contra indicated patients must be treated with hyperfractionated and/or accelerated treatment or a conventional radiotherapy combined with targeted therapy such as monoclonal antibody against the epidermal growth factor receptor. Supportive care is daily employed to increase the quality of life. PMID- 17137254 TI - [Post-therapeutic follow-up of head and neck cancer]. PMID- 17137255 TI - [Head and neck cancer in MEDLINE]. PMID- 17137256 TI - [Risk factors for head and neck cancer]. PMID- 17137257 TI - [Evidence-base medicine and critical appraisal]. PMID- 17137258 TI - [Suicidal behavior in the adolescent and in the adult]. PMID- 17137259 TI - [Problems posed by genetic diseases, concerning DNA instability disorders: fragile X syndrome]. PMID- 17137260 TI - [Acute retention of urine]. PMID- 17137261 TI - [Herpesvirus infections of the immunocompetent child and adult]. PMID- 17137262 TI - [Premature birth and fetal growth retardation]. PMID- 17137263 TI - [The Spanish influenza: a quickly forgotten viral pademic]. PMID- 17137264 TI - Pay and the gender gap: can the new system deliver for nurses? PMID- 17137265 TI - No substitute for nurses. AB - Field research in care homes has revealed for the first time the significant contribution that registered nurses make to patient care. PMID- 17137266 TI - Crisis in a drama. AB - Working as a nurse on movie and TV sets may involve international travel, excitement and adventure, but it is poorly paid and exhausting. PMID- 17137267 TI - Academic education's contribution to the nurse-patient relationship. AB - AIM: To explore how nursing students perceive the role of academic education in enabling them to understand factors that can influence the relationship between nurses and patients. METHOD: An exploratory qualitative design, with data collected through unstructured interviews with four pre-registration students enrolled on a degree course in adult nursing. FINDINGS: Participants emphasised the role of reflection in making academic education relevant to practice and in promoting understanding of the relationships they developed with patients. They also valued experiential learning methods used in seminars, for example, case studies, role play and group discussion. CONCLUSION: Academic education plays an important role in fostering understanding of the nurse-patient relationship by promoting a learning approach based on reflection and self-awareness. PMID- 17137268 TI - Psychological consequences of malignant melanoma: patients' experiences and preferences. AB - This article reports on the findings of a literature review exploring the psychological impact of malignant melanoma on patients, from diagnosis to follow up. It examines the preferences expressed by patients with regard to ongoing support from nurses in this field, and draws attention to some implications for practice. A small but significant body of literature provides information on the psychological stress experienced by patients with melanoma, and on their coping strategies. Preferences are expressed on the way that information is given at diagnosis and the support offered by the healthcare team during diagnosis and at later consultations. These preferences and experiences have implications for nurses working in cancer care in general, and skin cancer in particular. PMID- 17137269 TI - Chronic kidney disease: risk factors, assessment and nursing care. AB - This article provides an overview of chronic kidney disease in adults and outlines nursing management in primary and secondary care. Patient assessment, including the stages of chronic kidney disease, common causes and risk factors, are also discussed. PMID- 17137270 TI - Wound healing. PMID- 17137271 TI - Prison placement. PMID- 17137272 TI - Loss of experience. PMID- 17137273 TI - Release the tension. PMID- 17137274 TI - Scotland's staffing timebomb. PMID- 17137275 TI - Know the facts. AB - Vaccines to protect women against cervical cancer are already being licensed. An RCN leaflet gives nurses some of the facts about the links between HPV and cervical cancer. PMID- 17137276 TI - A change of heart. Interview by Carol Davis. AB - One Cambridge nurse has transformed the service for people needing elective cardioversion. Instead of complaints, the hospital now receives plaudits and its success has garnered an award. PMID- 17137277 TI - Raise awareness of stroke symptoms. AB - While many members of the public recognise that chest pain needs urgent attention, few people know much about stroke. Yet early treatment of stroke can reduce long-term disability. PMID- 17137278 TI - Let's visit the PROMs. PMID- 17137279 TI - Rapid response teams: a proactive strategy for improving patient care. AB - Cardiopulmonary resuscitation success rates have not changed in 30 years. Patient outcomes may improve if changes in a patient's condition are addressed at the onset of subtle deteriorations, rather than at the point of cardiac arrest. The rapid response team involves early intervention that demonstrates the ability to decrease cardiac arrest rates and improve patient mortality. PMID- 17137280 TI - Providing anticipatory care for patients with chronic obstructive pulmonary disease. AB - This article describes an initiative pioneered by the Met Office which uses weekly weather forecasts to enable the chronic obstructive pulmonary disease (COPD) outreach team in Plymouth to target patients most at risk of developing exacerbations. An active management plan is implemented during the winter to help prevent further deterioration in the patient's condition and possible hospital admissions. PMID- 17137281 TI - Strategies to explore innovation in nursing practice. AB - This article examines different types of innovation and explores how they can be applied to health care. It provides information on strategic that can be used by nurses to incorporate innovation in their practice. PMID- 17137282 TI - Reflection in practice. PMID- 17137283 TI - Sounds of silence. PMID- 17137284 TI - Defining moment. Interview by Drew Payne. PMID- 17137285 TI - Neurosurgical credentials: a history and critique. AB - The author presents a review of the process of peer evaluation in this century, as well as how certification and techniques of evaluation in the practice of medicine and neurosurgery have evolved. The effect of licensure on the workforce, the influence of trade laws on quality of education and training, and the new paradigm of competition and managed medical practices have made a remarkable impact on the nature of the care of patients in the United States. The author also discusses the effect of risk management and litigation on the quality of medical care, the nature of outcomes evaluation and economic credentials in the absence of appropriate instruments for such evaluation, and alternatives for the future of the credentials process. PMID- 17137286 TI - Whither neurosurgery? AB - Although trends in the marketplace demand for neurosurgeons should be of interest to neurosurgeons and prospective neurosurgeons, little data are available that accurately document these trends. A recent report published in the general medical journal Journal of the American Medical Association (JAMA) used the Conference Board help-wanted index to evaluate trends in physician marketplace demand. The authors of the JAMA study concluded that over the past 5 years there has been a significant fall in demand for specialist physicians. Because the discipline of neurosurgery was not included in the JAMA study, the authors of the present report attempt to evaluate the trend in the marketplace demand for neurosurgeons, using the same methodology of the JAMA study. The authors' data suggest that the conclusion of the JAMA study of steep declines in the demand for specialist physicians does not accurately reflect the job market for neurosurgeons, which in fact appears to be relatively stable. The present study attempts to document the stability of the neurosurgery market and outline the steps necessary to protect this market from existing threats. PMID- 17137287 TI - The Current Procedural Terminology and the Medicare Resource Based Relative Value Scale in neurosurgical practice. AB - Changes in healthcare have made it increasingly more important for neurosurgeons to understand the economic pressures that effect their reimbursement. Two fundamental concepts that are of the greatest importance are the Current Procedural Terminology coding and the Medicare Resource Based Relative Value Scale. The impact of these two entities on neurosurgical reimbursement in analyzed in this review. PMID- 17137288 TI - Medical case management after laminectomy or craniotomy: do all patients benefit from admission to the intensive care unit? AB - To define severity of illness to identify most effectively patients for whom admission to the intensive care unit (ICU) is unnecessary, the authors performed a retrospective cost-effectiveness analysis. The authors studied the records of 113 patients who were admitted to the ICU after undergoing laminectomy (or other spinal cord surgery) or craniotomy for removal of neoplasm; the Acute Physiology and Chronic Health Evaluation III prognostic system had identified these patients as having a 10% or less risk of requiring intervention while in the ICU. No patient required active intervention during a mean stay of 3.26 days in the ICU. Combined use of a "step-down" postoperative care unit and ICU can optimize allocation of medical resources while providing high-quality care for some neurosurgical patients who are at low risk of requiring postoperative intervention. PMID- 17137289 TI - Clinical practice guidelines and their role in quality of care assessments. AB - Clinical practice guidelines may be defined as systematically developed statements to assist practitioner and patient decisions about appropriate healthcare for specific clinical conditions. The practice guideline is a management plan that enables healthcare providers to make sequential decisions about testing or therapy in a given clinical situation. Given the heterogeneity of disease presentations and the complexity of making therapeutic decisions, the development of guidelines in neurosurgery presents unique problems and challenges. Clinical practice guidelines were developed in part as a method for the assessment of quality of medical care processes. Practice guidelines serve as a tool for comparisons to be made within and between healthcare delivery systems. The implementation of guidelines also allows for the acquisition of clinical and financial data that provide for outcomes analysis and other types of information. The characterization of patterns of care in different institutions and the relationship between patterns of care and outcomes will in the future allow for quality improvement, enable physicians to offer predictability and accountability to third-party interests, and contribute to an understanding of how to offer the best care to neurosurgical patients. The author provides an overview of clinical practice guidelines and illustrates their role in the assessment of quality of patient care, demonstrates how they fit into the methodology of quality assessments, and underscores the importances of practice quidelines for the development of disease management systems. PMID- 17137290 TI - The influence of secondary gain on surgical outcome: a comparison between cervical and lumbar discectomy. AB - Although the expectation of monetary compensation has been associated with failures in lumbar discectomy, the issue has not been investigated in patients undergoing cervical disc surgery. The authors analyzed the relationship between economic forms of secondary gain and surgical outcome in a group of patients with a common pay scale, retirement plan, and disability program. All procedures were performed at the Portsmouth Naval Medical Center between 1993 and 1995; active duty military servicepersons treated for cervical radiculopathy were prospectively included. Clinical, demographic, and financial factors were analyzed to determine which were predictive of outcome. Financial data were used to create a compensation incentive, which is proportional to the patient's rank, years of service, potential disability, retirement eligibility, and base pay and reflects the monetary incentive of disability. The results of cervical surgery were compared to a previously reported companion population of patients treated for lumbar disc disease. A good outcome was defined as a return to active duty, whereas a referral for disability was considered a poor surgical result. A 100% follow-up rate was obtained for 269 patients who underwent 307 cervical operations. Only 16% (43 of 269) of patients who underwent cervical operation received disability, whereas 24.7% (86 of 348) of patients who underwent lumbar discectomy obtained a poor result (p = 0.0082). Although economic forms of secondary gain were not associated with a poor outcome in cervical disease, both the rank (p = 0.002) and duration (p = 0.03) of an individual's military career were significant factors (p = 0.02). Of the medical variables tested, multilevel surgery (p = 0.03) and revision operations at the same level (p = 0.03) were associated with referral for medical discharge. Secondary gain in the form of economic compensation influences outcome in lumbar but not cervical disc surgery patients; the increased rate of disability referral in patients who underwent lumbar discectomy may reflect an expectation of economic compensation. Social factors that are independent of the anticipation of economic compensation seem to influence the outcome in cervical disc surgery patients. PMID- 17137291 TI - Physical interaction of two cancer-testis antigens, MAGE-C1 (CT7) and NY-ESO-1 (CT6). AB - Cancer/testis (CT) antigens are the protein products of germ line-associated genes that are activated in a wide variety of tumors and can elicit autologous cellular and humoral immune responses. CT antigens can be divided between those that are encoded on the X chromosome (CT-X antigens) and those that are not (non X CT antigens). Among the CT-X antigens, the melanoma antigen gene (MAGE) family, defined by a shared MAGE homology domain (MHD), is the largest. CT-X genes are frequently expressed in a coordinate manner in cancer cells, and their expression appears to be modulated by epigenetic mechanisms. The expression of CT-X genes is associated with advanced disease and poor outcome in different tumor types. We used the yeast two-hybrid system to identify putative MHD-interacting proteins. The MHD of MAGE-C1 (CT7) was used as bait to screen a human testis cDNA library. This study identified NY-ESO-1 (CT6) as a MAGE-C1 binding partner. Immunoprecipitation and immunofluorescence staining confirmed MAGE-C1 interaction with NY-ESO-1, and cytoplasmic co-localization of both proteins in melanoma cells. Co-expression of these two genes was found to occur in cancer cell lines from different origins, as well as in primary tumors (multiple myeloma and non small cell lung cancer samples). This is the first report of direct interaction between two CT antigens and may be pertinent in the light of the frequently coordinated expression of these proteins. PMID- 17137292 TI - Process technology for production and recovery of heterologous proteins with Pichia pastoris. AB - Developments in process techniques for production and recovery of heterologous proteins with Pichia pastoris are presented. Limitations for the standard techniques are described, and alternative techniques that solve the limitations problems are reviewed together with the methods that resulted in higher productivity of the P. pastoris processes. The main limitations are proteolysis of the secreted products and cell death in the high cell density bioreactor cultures. As a consequence, both low productivity and lower quality of the feedstock for downstream processing are achieved in processes hampered with these problems. Methods for exploring proteolysis and cell death are also presented. Solving the problems makes the conditions for downstream processing superior for the P. pastoris expression systems compared to other systems, which either need complex media or rely on intracellular production. These improved conditions allow for interfacing of cultivation with downstream processing in an integrated fashion. PMID- 17137294 TI - Microalgal reactors: a review of enclosed system designs and performances. AB - One major challenge to industrial microalgal culturing is to devise and develop technical apparata, cultivation procedures and algal strains susceptible of undergoing substantial increases in efficiency of use of solar energy and carbon dioxide. Despite several research efforts developed to date, there is no such thing as "the best reactor system"- defined, in an absolute fashion, as the one able to achieve maximum productivity with minimum operation costs, irrespective of the biological and chemical system at stake. In fact, choice of the most suitable system is situation-dependent, as both the species of alga available and the final purpose intended will play a role. The need of accurate control impairs use of open-system configurations, so current investigation has focused mostly on closed systems. In this review, several types of closed bioreactors described in the technical literature as able to support production of microalgae are comprehensively presented and duly discussed, using transport phenomenon and process engineering methodological approaches. The text is subdivided into subsections on: reactor design, which includes tubular reactors, flat plate reactors and fermenter-type reactors; and processing parameters, which include gaseous transfer, medium mixing and light requirements. PMID- 17137293 TI - From 3D to 2D: a review of the molecular imprinting of proteins. AB - Molecular imprinting is a generic technology that allows for the introduction of sites of specific molecular affinity into otherwise homogeneous polymeric matrices. Commonly this technique has been shown to be effective when targeting small molecules of molecular weight <1500, while extending the technique to larger molecules such as proteins has proven difficult. A number of key inherent problems in protein imprinting have been identified, including permanent entrapment, poor mass transfer, denaturation, and heterogeneity in binding pocket affinity, which have been addressed using a variety of approaches. This review focuses on protein imprinting in its various forms, ranging from conventional bulk techniques to novel thin film and monolayer surface imprinting approaches. PMID- 17137295 TI - A computational procedure for optimal engineering interventions using kinetic models of metabolism. AB - The identification of optimal intervention strategies is a key step in designing microbial strains with enhanced capabilities. In this paper, we propose a general computational procedure to determine which genes/enzymes should be eliminated, repressed or overexpressed to maximize the flux through a product of interest for general kinetic models. The procedure relies on the generalized linearization of a kinetic description of the investigated metabolic system and the iterative application of mixed-integer linear programming (MILP) optimization to hierarchically identify all engineering interventions allowing for reaction eliminations and/or enzyme level modulations. The effect of the magnitude of the allowed changes in concentrations and enzyme levels is investigated, and a variant of the method to explore high-fold changes in enzyme levels is also analyzed. The proposed framework is demonstrated using a kinetic model modeling part of the central carbon metabolism of E. coli for serine overproduction. The proposed computational procedure is a general approach that can be applied to any metabolic system for which a kinetic description is provided. PMID- 17137296 TI - High-level gene expression for recombinant penicillin acylase production using the araB promoter system in Escherichia coli. AB - The pac gene encoding penicillin acylase (PAC) was overexpressed under the regulation of the araB promoter (ParaB, also known as PBAD) in Escherichia coli (E. coli). The current ParaB expression system exhibited minimum leaking pac expression in the absence of arabinose as well as fast and high-level pac expression upon induction with arabinose in a wide concentration range. The production of PAC was limited by the accumulation of PAC precursors (i.e., proPAC in both soluble and insoluble forms) and various negative cellular responses, such as growth arrest and cell lysis. The culture performance could be improved by degP coexpression and the individual contribution of DegP protease and chaperone activities to the enhancement on the production of PAC was characterized. The study highlights the importance of identifying the step(s) limiting high-level gene expression and subsequent design and construction of the host/vector system for enhancing recombinant protein production in E. coli. PMID- 17137297 TI - Phytotransformation of 2,4-dinitrotoluene in arabidopsis thaliana: toxicity, fate, and gene expression studies in vitro. AB - Basic knowledge of the plant transformation pathways and toxicity of 2,4 dinitrotoluene (2,4-DNT) will assist in the design and assessment of a phytoremediation strategy. This study presents the toxicity and fate of 2,4-DNT and gene expression in response to 2,4-DNT exposure using the model plant Arabidopsis thaliana, an increasingly popular system for genetic and biochemical studies of phytotransformation of explosives. From the results of biomass and root growth assays for toxicity, 2,4-DNT was toxic to the plants at concentrations as low as 1 mg/L. In the uptake study, 95% of the initial 2,4-DNT was removed by 15-day-old seedlings from liquid media regardless of the initial 2,4-DNT concentrations while 30% accounted for the adsorption to the autoclaved plant materials. The mass balance was over 86% using [U-14C]2,4-DNT, and the mineralization by the plants was less than 1% under sterile conditions during 14 days of exposure. The percentage of the bound radioactivity increased from 49% to 72% of the radioactivity in the plants, suggesting transformed products of 2,4 DNT may be incorporated into plant tissues such as lignin and cellulose. Monoaminonitrotoluene isomers and unknown metabolites with short retention times were detected as transformed products of 2,4-DNT by the plants. Most (68%) of the radioactivity taken up by the plants was in the root tissues in nonsterile hydroponic cultures. Glutathione and expression of related genes (GSH1 and GSH2) in plants exposed to 2,4-DNT were 1.7-fold increased compared to untreated plants. Genes of a glutathione S-transferase and a cytochrome P450, which were induced by 2,4,6-trinitrotoluene exposure in previous studies, were upregulated by 10- and 8-fold, respectively. The application of phytoremediation and the development of transgenic plants for 2,4-DNT may be based on TNT phytotransformation pathway characteristics because of the similar fate and gene expression in plants. PMID- 17137298 TI - Modeling culture profiles of the heterocystous N2-fixing cyanobacterium Anabaena flos-aquae. AB - Heterocyst differentiation is a unique feature of nitrogen-fixing cyanobacteria, potentially important for photobiological hydrogen production. Despite the significant advances in genetic investigation on heterocyst differentiation, there were no quantitative culture-level models that describe the effects of cellular activities and cultivation conditions on the heterocyst differentiation. Such a model was developed in this study, incorporating photosynthetic growth of vegetative cells, heterocyst differentiation, self-shading effect on light penetration, and nitrogen fixation. The model parameters were determined by fitting experimental results from the growth of the heterocystous cyanobacterium Anabaena flos-aquae CCAP 1403/13f in media without and with different nitrate concentrations and under continuous illumination of white light at different light intensities (2, 5, 10, 17, 20 and 50 microE m-2 s-1). The model describes the experimental profiles well and gives reasonable predictions even for the transition of growth from that on external N source to that via nitrogen fixation, responding to the change in external N concentrations. The significance and implications of the best-fit values of the model parameters are discussed. PMID- 17137299 TI - Metabolic engineering and characterization of phaC1 and phaC2 genes from Pseudomonas putida KCTC1639 for overproduction of medium-chain-length polyhydroxyalkanoate. AB - Two PHA synthase phaC1 and phaC2 genes cloned from the new strain Pseudomonas putida KCTC1639 were metabolically engineered for the overproduction of medium chain-length polyhydroxyalkanoate (mcl-PHA). The overexpressed phaC1 and phaC2 genes in P. putida KCTC1639 were compared in terms of the biosynthesis of mcl PHA, fatty acid assimilation, distribution of 3-hydroxylacyl monomer units, granular morphology, and thermophysical properties of the accumulated mcl-PHA. The biosynthesis of mcl-PHA was enhanced only by the overexpressed phaC1 gene up to 2.86-fold, in contrast, the phaC2 gene did not activate the biosynthesis of mcl-PHA. The overexpressed phaC1 gene tended to form enlarged, high molecular weight, and lower crystalline mcl-PHA granules, whereas the amplified phaC2 gene induced the fragmentation of mcl-PHA into a few small-sized granules. The transformant P. putida KCTC1639 overexpressing the phaC1 gene encoding PHA synthase I was cultivated by pH-stat fed-batch cultivation, and the concentration and content of mcl-PHA increased up to 8.91 g L-1 and 70.5%, respectively. PMID- 17137300 TI - Effect of glucose or glycerol as the sole carbon source on gene expression from the Salmonella prpBCDE promoter in Escherichia coli. AB - We have developed an expression system (Salmonella-based pPro system) containing the Salmonella enterica prpBCDE promoter (PprpB) and prpR encoding the positive transcriptional regulator of this promoter. In this study, the transcriptional efficiency of the pPro expression system was measured by placing the gene encoding the green fluorescent protein (gfp) under the control of PprpB and growing cells containing this construct in minimal medium supplemented with glucose or glycerol as a sole carbon source. In wild-type Escherichia coli (E. coli) BL21, the system exhibited high induced expression as well as high background expression; however, in E. coli JSB, a sbm-ygfDGHI deletion mutant of E. coli BL21(DE3), the system showed low background expression and high induced expression. The system exhibited homogeneous expression at the single-cell level, highly regulatable expression over a wide range of propionate concentrations, and fully induced expression at a low propionate concentration relative to that needed to induce the system in rich, undefined medium. The expression system is comparable to the widely used T7 promoter-driven expression systems in glucose or glycerol minimal medium. PMID- 17137301 TI - Removal of high phenol concentrations with adapted activated sludge in suspended form and entrapped in calcium alginate /cross-linked poly(N-vinyl pyrrolidone) hydrogels. AB - The present work evaluates the aerobic removal of 0.25-2 g/L of phenol by adapted activated sludge in batch and continuous reactors, in suspended form and trapped in polymeric hydrogel beads of calcium alginate(1%) and cross-linked poly(N-vinyl pyrrolidone), x-PVP (4%). The mechanical and chemical resistance of the entrapping hydrogel was also evaluated in three different media: (I) rich in phosphate and ammonium ions; (II) using alternate P and N sources, and (III) without nutrients. The adapted consortium removed phenol concentrations up to 2 g/L more efficiently in the immobilized systems. A decrease in phenol removal rate was observed as the food/microorganisms (F/M) ratio increased. A zero-order kinetics was observed with phenol concentrations > 1 g/L and a first-order kinetics at concentrations < 1 g/L. The best response (100% removal) was in the continuous reactors using type II medium, with a hydraulic residence time (HRT) of 12.5 h, an influent pH = 5, and an F/M ratio below 0.25. The immobilizing matrix deteriorated after 170 h of use in continuous reactors, especially with media I and II, probably due to the attrition forces, to chemical weakness of the material, and to the pressure of the bacterial growth inside the bead. PMID- 17137302 TI - Development and analysis of a mathematical model for antibody-producing GS-NS0 cells under normal and hyperosmotic culture conditions. AB - The GS-NS0 cell line is industrially important and is currently used for the large-scale production of several therapeutic monoclonal antibodies. A novel hybrid model, consisting of both unstructured and structured elements, has been developed to describe cell growth and death, metabolism, and antibody production in the GS-NS0 system under normal culture conditions. A comparison between the hybrid model and a large-scale single-cell model (SCM) describing detailed metabolic processes verified the predictive ability of the hybrid model (when compared with experimental data) and highlighted the practical difficulties involved in utilizing complex models. Global sensitivity analysis (GSA) on the hybrid model identified the specific transcription and translation rates of heavy and light immunoglobulin chains as parameters with the largest impact on the antibody production process. This information, together with the addition of a 24 h lag phase, resulted in the successful extension of the hybrid model to represent GS-NS0 system behavior under hyperosmotic culture conditions. PMID- 17137303 TI - Cell-free protein synthesis system prepared from insect cells by freeze-thawing. AB - We established a novel cell-free protein synthesis system derived from Trichoplusia ni (HighFive) insect cells by a simple extraction method. Luciferase and beta-galactosidase were synthesized in this system with active forms. We analyzed and optimized (1) the preparation method of the insect cell extract, (2) the concentration of the reaction components, and (3) the 5'-untranslated region (5'-UTR) of mRNA. The extract was prepared by freeze-thawing insect cells suspended in the extraction buffer. This preparation method was a simple and superior method compared with the conventional method using a Dounce homogenizer. Furthermore, protein synthesis efficiency was improved by the addition of 20% (v/v) glycerol to the extraction buffer. Concentrations of the reaction components were optimized to increase protein synthesis efficiency. Moreover, mRNAs containing 5'-UTRs derived from baculovirus polyhedrin genes showed high protein synthesis activity. Especially, the leader composition of the Ectropis obliqua nucleopolyhedrovirus polyhedrin gene showed the highest enhancement activity among the six 5'-UTRs tested. As a result, in a batch reaction approximately 71 microg of luciferase was synthesized per milliliter of reaction volume at 25 degrees C for 6 h. Moreover, this method for the establishment of a cell-free system was applied also to Spodoptera frugiperda 21 (Sf21) insect cells. After optimizing the concentrations of the reaction components and the 5' UTR of mRNA, approximately 45 microg/mL of luciferase was synthesized in an Sf21 cell-free system at 25 degrees C for 3 h. These productivities were sufficient to perform gene expression analyses. Thus, these cell-free systems may be a useful tool for simple synthesis in post-genomic studies as a novel protein production method. PMID- 17137304 TI - Enantioselective synthesis of L-homophenylalanine by whole cells of recombinant Escherichia coli expressing L-aminoacylase and N-acylamino acid racemase genes from Deinococcus radiodurans BCRC12827. AB - L-Homophenylalanine (l-HPA) is a chiral unnatural amino acid used in the synthesis of angiotensin converting enzyme inhibitors and many pharmaceuticals. To develop a bioconversion process with dynamic resolution of N-acylamino acids for the l-HPA production, N-acylamino acid racemase (NAAAR) and l-aminoacylase (LAA) genes were cloned from Deinococcus radiodurans BCRC12827 and expressed in Escherichia coli XLIBlue. The recombinant enzymes were purified by nickel-chelate chromatography, and their biochemical properties were determined. The NAAAR had high racemization activity toward chiral N-acetyl-homophenylalanine (NAc-HPA). The LAA exhibited strict l-enantioselection to hydrolyze the NAc-l-HPA. A stirred glass vessel containing transformed E. coli cells expressing D. radiodurans NAAAR and LAA was used for the conversion of NAc-d-HPA to l-HPA. Unbalance activities of LAA and NAAAR were found in E. coli cell coexpressing laa and naaar genes, which resulted in the accumulation of an intermediate, NAc-l-HPA, in the early stage of conversion and a low productivity of 0.83 mmol l-HPA/L h. The results indicated that low activity of LAA present in the biomass is the rate-limiting factor in l-HPA production. In the case of two whole cells with separately expressed enzyme, the enzymatic activities of LAA and NAAAR could be balanced by changing the loading of individual cells. When the activities of two enzymes were fixed at 3600 U/L, 99.9% yield of l-HPA could be reached in 1 h, with a productivity of 10 mmol l-HPA/L h. The cells can be reused at least six cycles at a conversion yield of more than 96%. This is the first NAAAR/LAA process using NAc-HPA as substrate and recombinant whole cells containing Deinococcus enzymes as catalysts for the production of l-HPA to be reported. PMID- 17137305 TI - Preparation and characterization of urea-formaldehyde-pepsin bioconjugate: a new biocatalyst system. AB - This study describes the synthesis of urea formaldehyde (UF) microspheres by a dispersion polycondensation polymerization method. These microspheres with proper F/U molar ratio can provide highly reactive groups, capable of further condensation with the amino acid residues of enzyme/proteins. Presence of methylols groups in UF microspheres was confirmed by 13C NMR study. Pepsin, a proteolytic enzyme, was immobilized on the UF microspheres to form bioconjugate system. As compared to the free enzyme in solution, the pepsin in the bioconjugate system exhibited significantly enhanced pH and temperature stability. The urea-formaldehyde-pepsin bioconjugate system also exhibited excellent proteolytic activity over eight successive reuse cycles with more than 50% of initial activity. A highlight of this new biocatalyst is the ease with which separation of this biocatalyst from the reaction medium may be achieved by mild centrifugation. PMID- 17137306 TI - High concentration cultivation of Bifidobacterium bifidum in a submerged membrane bioreactor. AB - In batch cultures, after 25 h, the maximum cell mass of Bifidobacterium bifidum BGN4 was 4.5 g/L, and the maximum cell count was 3.0 x 10(9) cfu/mL at pH 6.0 and 50 g/L sucrose. To increase the viable counts of bifidobacteria, cell retentive culture was applied using a submerged membrane bioreactor with suction and gas sparging. The maximum mass, count, and productivity of the cells after 36 h were 12.0 g/L, 2.2 x 10(10) cfu/mL, and 6.1 x 10(8) cfu/mL x h, respectively, at the feeding (dilution) rate of 120 mL/h (0.06 h-1) in the feeding medium. The accumulated levels of organic acids and ammonium ions at the end of the cultivation were 1.5 and 1.0 g/L, respectively. The viable counts and volumetric productivity of the cells after the cell retentive culture were 7.3- and 5.1-fold higher, respectively, than the values obtained during batch culture. These high viable counts and volumetric productivities were obtained by maintaining lower concentrations of organic acids and ammonium ions so that the growth of B. bifidum BGN4 was not inhibited. The submerged membrane bioreactor produced the highest viable counts of B. bifidum without membrane fouling and cell damage. PMID- 17137307 TI - Enhancement of cellulose pellicle production by constitutively expressing vitreoscilla hemoglobin in Acetobacter xylinum. AB - Vitreoscilla hemoglobin (VHb) gene driven by the constitutive bla promoter was expressed in the cellulose-producing Acetobacter xylinum. The expressed VHb was biochemically active and could enhance cell growth in a shaken culture containing cellulase. VHb-expressing A. xylinum (VHb+) exhibited a specific growth rate 50% higher than that of the host strain (VHb-). Probably because of its faster growth rate, the size of tentacled cellulose beads produced by VHb+ was about 20% of that produced by VHb- after 2 days cultivation in a shake-flask. When cultured statically, the amount of cellulose pellicle produced by VHb+ could be 2-fold that produced by VHb-. Cellulose pellicle concentration of 11 g/L was obtained for VHb+, whereas 6 g/L was obtained for VHb- after 6 days of microaerobic incubation. PMID- 17137308 TI - Recovery of rhodium(III) from solutions and industrial wastewaters by a sulfate reducing bacteria consortium. AB - A quantitative analysis of the rate of removal of rhodium(III) by a resting sulfate-reducing bacteria (SRB) consortium under different initial rhodium and biomass concentrations, pH, temperature, and electron donor was studied. Rhodium speciation was found to be the main factor controlling the rate of its removal from solution. SRB cells were found to have a higher affinity for anionic rhodium species, as compared to both cationic and neutral species, which become abundant when speciation equilibrium was reached. Consequently, a pH-dependent rate of rhodium removal from solution was observed. The maximum SRB uptake capacity for rhodium was found to be 66 mg of rhodium per gram of resting SRB biomass. Electron microscopy studies revealed a time-dependent localization and distribution of rhodium precipitates, initially intracellularly and then extracellularly, suggesting the involvement of an enzymatic reductive precipitation process. When a purified hydrogenase enzyme was incubated with rhodium chloride solution under hydrogen, 88% of the rhodium was removed within 1 h, whereas with a soluble extract from SRB 77% was removed within 10 min. Due to the low pH of the industrial effluent (1.31), the enzymatic reduction of rhodium by the purified hydrogenase was greatly limited, and it was apparent that an industrial effluent pretreatment was necessary before the application of an enzymatic treatment. In the present study, however, it was established that SRB are good candidates for the enzymatic recovery of rhodium from both aqueous solution and industrial effluent. PMID- 17137309 TI - The integrated simulation and assessment of the impacts of process change in biotherapeutic antibody production. AB - Growing commercial pressures in the pharmaceutical industry are establishing a need for robust computer simulations of whole bioprocesses to allow rapid prediction of the effects of changes made to manufacturing operations. This paper presents an integrated process simulation that models the cGMP manufacture of the FDA-approved biotherapeutic CroFab, an IgG fragment used to treat rattlesnake envenomation (Protherics U.K. Limited, Blaenwaun, Ffostrasol, Llandysul, Wales, U.K.). Initially, the product is isolated from ovine serum by precipitation and centrifugation, before enzymatic digestion of the IgG to produce FAB and FC fragments. These are purified by ion exchange and affinity chromatography to remove the FC and non-specific FAB fragments from the final venom-specific FAB product. The model was constructed in a discrete event simulation environment and used to determine the potential impact of a series of changes to the process, such as increasing the step efficiencies or volumes of chromatographic matrices, upon product yields and process times. The study indicated that the overall FAB yield was particularly sensitive to changes in the digestive and affinity chromatographic step efficiencies, which have a predicted 30% greater impact on process FAB yield than do the precipitation or centrifugation stages. The study showed that increasing the volume of affinity matrix has a negligible impact upon total process time. Although results such as these would require experimental verification within the physical constraints of the process and the facility, the model predictions are still useful in allowing rapid "what-if" scenario analysis of the likely impacts of process changes within such an integrated production process. PMID- 17137310 TI - Purification of RT-PCR competent poly(A) mRNA from crude cell lysate by affinity precipitation. AB - Stimuli-responsive bioconjugates consisting of avidin covalently linked to poly(N isopropylacrylamide) were used for the recovery of poly(A) mRNA hybridized to biotinylated poly(dT)-tags from crude cell lysates (Jurkat cells) by affinity precipitation. The bioconjugates are soluble in cold water but precipitate readily once a critical solution temperature (33 degrees C in pure water) is surpassed. The process is fully reversible and shows the expected dependencies on the composition of the aqueous solution and the bioconjugate chemistry. The results of the affinity precipitation were compared to those achieved with an accepted standard purification of poly(A) mRNA using avidin-activated magnetic beads. Both yield and quality/purity of the affinity precipitated poly(A) mRNA were found to be similar or better (especially removal of rRNA) than for poly(A) mRNA prepared by the magnetic particle-based protocol, while both mRNA isolates performed equally well in standard reverse transcriptase amplification (RT-PCR) of a beta actin transcript fragment. Poly(A) mRNA purification schemes based on affinity precipitation require no dedicated equipment and should have advantages in terms of scalability, handling, and costs. PMID- 17137311 TI - Medium term planning of biopharmaceutical manufacture with uncertain fermentation titers. AB - The growing trend of employing multiproduct manufacturing facilities along with the randomness inherent in the biopharmaceutical manufacturing environment is creating significant scheduling and planning challenges for the biopharmaceutical industry. This work focuses on capturing the effect of uncertainty in fermentation titers when optimizing the planning of biopharmaceutical manufacturing campaigns. A mixed integer linear programming (MILP) model based on previous work is derived via chance constrained programming (CCP). The methodology is applied to two illustrative examples, and the results are compared with those from the deterministic model and a multiscenario model accompanied by an iterative construction algorithm. The computational results indicate that the proposed methodology offers significant improvements in solution quality over the compared approaches and presents an opportunity for biopharmaceutical manufacturers to make better medium term planning decisions, particularly under uncertain manufacturing conditions. PMID- 17137312 TI - Kinetic and thermodynamic investigation on ascorbate oxidase activity and stability of a Cucurbita maxima extract. AB - The kinetic and thermodynamic properties of ascorbate oxidase (AO) activity and stability of a Cucurbita maxima extract were investigated. Activity tests performed at 25 degrees C using initial ascorbic acid concentration in the range 50-750 M allowed estimating the Michaelis constant for this substrate (Km = 126 microM) and the maximum initial rate of ascorbic acid oxidation (A0,max = 1.57 mM min-1). The main thermodynamic parameters of the enzyme reaction (DeltaH* = 10.3 kJ mol-1; DeltaG* = 87.2 kJ mol-1; DeltaS* = -258 J mol-1 K-1) were estimated through activity tests performed at 25-48 C. Within such a temperature range, no decrease in the initial reaction rate was detected. The long-term thermostability of the raw extract was then investigated by means of residual activity tests carried out at 10-70 degrees C, which allowed estimating the thermodynamic parameters of the irreversible enzyme inactivation as well (DeltaH*D = 51.7 kJ mol-1; DeltaG*D = 103 kJ mol-1; S*D = -160 J mol-1 K-1). Taking into account the specific rate of AO inactivation determined at different temperatures, we also estimated the enzyme half-life (1047 min at 10 degrees C and 21.2 min at 70 degrees C) and predicted the integral activity of a continuous system using this enzyme preparation. This work should be considered as a preliminary attempt to characterize the AO activity of a C. maxima extract before its concentration by liquid-liquid extraction techniques. PMID- 17137313 TI - Production, secretion, and stability of human secreted alkaline phosphatase in tobacco NT1 cell suspension cultures. AB - Tobacco NT1 cell suspension cultures secreting active human secreted alkaline phosphatase (SEAP) were generated for the first time as a model system to study recombinant protein production, secretion, and stability in plant cell cultures. The SEAP gene encodes a secreted form of the human placental alkaline phosphatase (PLAP). During batch culture, the highest level of active SEAP in the culture medium (0.4 U/mL, corresponding to approximately 27 mg/L) was observed at the end of the exponential growth phase. Although the level of active SEAP decreased during the stationary phase, the activity loss did not appear to be due to SEAP degradation (based on Western blots) but due to SEAP denaturation. The protein stabilizing agents polyvinylpirrolidone (PVP) and bacitracin were added extracellularly to test for their ability to reduce the loss of SEAP activity during the stationary phase. Bacitracin (100 mg/L) was the most effective treatment at sustaining activity levels for up to 17 days post-subculture. Commercially available human placental alkaline phosphatase (PLAP) was used to probe the mechanism of SEAP deactivation. Experiments with PLAP in sterile and conditioned medium corroborated the denaturation of SEAP by factors generated by cell growth and not due to simple proteolysis. We also show for the first time that the factors promoting activity loss are heat labile at 95 degrees C but not at 70 degrees C, and they are not inactivated after a 5 day incubation period under normal culture conditions (27 degrees C). In addition, there were no significant changes in pH or redox potential when comparing sterile and cell-free conditioned medium during PLAP incubation, indicating that these factors were unimportant. PMID- 17137314 TI - Investigation of bacteriophage MS2 viral dynamics using model discrimination analysis and the implications for phage therapy. AB - Lytic phages infect their bacterial hosts, use the host machinery to replicate, and finally lyse and kill their hosts, releasing progeny phages. Various mathematical models have been developed that describe these phage-host viral dynamics. The aim of this study was to determine which of these models best describes the viral dynamics of lytic RNA phage MS2 and its host Escherichia coli C-3000. Experimental data consisted of uninfected and infected bacterial cell densities, free phage density, and substrate concentration. Parameters of various models were either determined directly through other experimental techniques or estimated using regression analysis of the experimental data. The models were evaluated using a Bayesian-based model discrimination technique. Through model discrimination it was shown that phage-resistant cells inhibited the growth of phage population. It was also shown that the uninfected bacterial population was a quasispecies consisting of phage-sensitive and phage-resistant bacterial cells. When there was a phage attack the phage-sensitive cells died out and the phage resistant cells were selected for and became the dominant strain of the bacterial population. PMID- 17137315 TI - Determination of biomass composition of Catharanthus roseus hairy roots for metabolic flux analysis. AB - Metabolic flux analysis is a powerful diagnostic tool in metabolic engineering, and determination of biomass composition is indispensable to accurate flux evaluation. We report the elemental and biomolecular composition of Catharanthus roseus hairy roots, a pharmaceutically significant plant system and an important metabolic engineering target. The molecular formula of the organic material in the hairy roots was C12.0H22.7N0.4O7.6 during mid-exponential growth. The abundances of lipids, lignin, cellulose, hemicellulose, starch, protein, proteinogenic amino acids, mineral ash, and moisture in the biomass were quantified. Analysis of water-soluble components of the biomass with 1-D 13C and 2-D [1H,1H] correlation (COSY) NMR spectroscopy revealed that the water-soluble components were composed almost entirely of -glucans. Agropine, a frequently reported hairy root biomass component, was not detected. Our measurements of the biomass components quantified 83.6 +/- 9.3% (w/w) of the biomass. Together with previously reported abundances of indole alkaloids, we accounted for at least 85.9 +/- 11.6% (w/w) of the carbon in the biomass, which enabled the precise determination of 12 biomass synthesis fluxes. PMID- 17137316 TI - Effects of composition, solvent, and salt particles on the physicochemical properties of polyglycolide/poly(lactide-co-glycolide) scaffolds. AB - Polyglycolide (PGA)/poly(lactide-co-glycolide) (PLGA) scaffolds were fabricated by a solvent casting/particulate leaching method using hexafluoroisopropanol (HFIP) or acetone for material dissolution and NaCl particles as porogen. The results revealed that the mechanical strength increased as the PGA percentage in a HFIP-processed scaffold increased. Chemical ingredients did not substantially affect the mechanical strength of acetone-processed scaffolds. Large NaCl particles led to weak mechanical strength, low porosity, and small specific surface area. For a fixed composition, PGA crystals in a HFIP-processed scaffold were smaller than those in an acetone-processed scaffold. High PGA fractions yielded partly fused PGA/PLGA scaffolds. A faster degradation rate of a scaffold could result from a higher PGA percentage, smaller NaCl particles, or the existence of chondrocytes. The combination of PGA and PLGA, which compensated each other for bioactivity, would be beneficial to cartilage regeneration. PMID- 17137317 TI - Development of a culture sub-population induction model: signaling pathways synergy and taxanes production by Taxus canadensis. AB - Cell cultures of Taxus canadensis were subjected to exogenously applied ethylene (ET) hormone and methyl jasmonate (MJ) elicitation in factorial design experiments. Levels of extracellular taxanes, including paclitaxel, were used with principal component analysis for fault detection and real-coded genetic algorithms for parameter optimization to construct a culture sub-population induction model. Culture sub-populations were identified by the model as (1) uninduced, (2) induced to unilateral function of the ET-signaling pathway, and (3) induced to cooperation between jasmonic acid (JA)- and ET-signaling pathways. Comprehensive model results suggested greater rates of cellular induction (resulting in exogenous taxane production) by ET gas as opposed to MJ elicitation. However, cellular induction of ET-signaling pathway genes increased the rate of induction of JA-signaling pathway genes by orders of magnitude. In addition, model results showed that induction of genes leading to extracellular production of the simple taxane 10-deacetylbaccatin III was regulated by the unilateral ET-signaling pathway. However, it was suggested that further processing of this simple taxane to complex taxane structures, such as paclitaxel, required further gene induction by the JA-signaling pathway. Thus, production rate constants of exogenous complex taxanes were predicted to be an order of magnitude lower than that for the simple taxane 10-deacetylbaccatin III. The fraction of the cell culture sub-population displaying unilateral ET signaling pathway gene induction was found inversely proportional to levels of MJ elicitation. When coupled with simple non-growth product models, levels of all extracellular taxanes were effectively predicted using the culture sub-population induction model. PMID- 17137318 TI - Culture of neural stem cells in calcium alginate beads. AB - Neural stem cells (NSCs) with the capacity of extensive self-renewal and multilineage differentiation have attracted more and more attention in research as NSCs will play an important role in the nerve disease treatment and nerve injury repair. The shortage of NSCs, both their sources and their numbers, however, is the biggest challenge for their clinic application, and hence, in vitro culture and expansion of NSCs is vitally important to realize their potentials. In this work, mouse-derived NSCs were cultured in three-dimensional calcium alginate beads (Ca-Alg-Bs). Gelling conditions, cell density, and cell harvest were determined by the exploration of formation and dissociation parameters for Ca-Alg-Bs. Additionally, the recovered and the subsequent induced cells were identified by immunofluorescence staining of Nestin, beta-tubulin, and GFAP. The results show that the 2-mm diameter Ca-Alg-Bs, prepared with 1.5% sodium alginate solution and 3.5% CaCl2 solution and with gelling for 10 min, is suitable for the NSCs culture. The seeding density of 0.8 x 10(5) cells x mL-1 for the encapsulation of NSCs resulted in the most expansion, and the NSCs almost doubled during the experiment. The average cell recovery rate is over 88.5%, with the Ca-Alg-Bs dissolving in 55 mM sodium citrate solution for 10 min. The recovered cells cultured in the Ca-Alg-Bs still expressed Nestin and had the capacity of multilineage differentiation into neurons and glial cells and, thus, remained to be NSCs. These results demonstrate that NSC expansion within Ca-Alg Bs is feasible and provides further possibilities for NSC expansion in bioreactors of the scale of clinical relevance. PMID- 17137319 TI - Greenbottle (Lucilia sericata) larval secretions delivered from a prototype hydrogel wound dressing accelerate the closure of model wounds. AB - The resurgence of larval biotherapy as a debridement tool in wound management has been accompanied by several clinical reports highlighting concomitant tissue regeneration. Studies employing in vitro cell motility assays have found that purified excretory/secretory (ES) products from Greenbottle larvae (blowfly, Lucilia sericata) are motogenic for human dermal fibroblasts when used as a supplement in culture media. The objective of the present study was to determine whether ES delivered using a prototype hydrogel wound dressing induced similar motogenic effects on fibroblastic (3T3) and epithelial cells (HaCaTs) comprising a scratched-monolayer wound model. Quantitative analysis by MTT assay failed to detect significant mitogenic effects of ES on either cell type. Quantitative image analysis revealed that ES exposure markedly accelerated wound closure through a motogenic effect on both fibroblasts and keratinocytes. Quantitative histochemical analysis detected significantly higher phosphotyrosine (pTyr) expression in ES-exposed cell cultures than in controls; moreover immunocytochemistry revealed conspicuously raised levels of pTyr expression in cells located at the wound margin. By attenuation with a panel of enzyme inhibitors these effects were attributed to the protease components of ES. The present results suggest that controlled delivery of ES as a follow-up to maggot debridement therapy may be an effective therapeutic option for stimulation of tissue regeneration in wound management. PMID- 17137320 TI - Examination of mineralized nodule formation in living osteoblastic cultures using fluorescent dyes. AB - Detecting the formation of mineralized nodules in osteogenic cell culture provides a means of assessing mature osteoblast cell function and the status of culture. In the present study, to continuously monitor the formation of mineralized nodules during the entire culture period, different concentrations of two fluorescent dyes (xylenol orange and calcein blue) were evaluated for their ability to specifically label calcified areas and their toxicity to cells in osteogenic cultures. Results showed that 20 microM xylenol orange and 30 microM calcein blue gave rise to distinct fluorescent staining for mineralized nodules, which were correlated exactly with von Kossa and alizarin red S staining at the same locations in cultures. In the assessment of toxicity, both dyes at the aforementioned concentrations did not alter cell viability or change the total DNA content in cultures. To demonstrate the advantage of using these fluorochromes to monitor mineralized nodules formation, consecutive fluorescent images of each staining were recorded at the same location of individual culture over the entire duration. The result indicates that both xylenol orange and calcein blue can provide contrasting fluorescent staining to continuously monitor mineralized nodules formation in living osteogenic cell cultures without deleterious effects. PMID- 17137321 TI - Stability of green fluorescent protein (GFP) in chlorine solutions of varying pH. AB - Green fluorescent protein (GFP) is an excellent biosensor as a result of its ability to be easily monitored in a wide variety of applications. Enzymes and proteins have been used as biological indicators to evaluate the immediate efficacy of industrial procedures, such as blanching, pasteurization, and disinfection treatments, as well as to monitor the satisfactory preservation of a product subjected to disinfection or sterilization. The purpose of this work was to study GFP stability in chlorinated water for injection (WFI) and chlorinated buffered solutions at various pH ranges, with and without agitation, to evaluate the exposure time required for chlorine to decrease 90% of its fluorescence intensity (decimal reduction time, D-value, min, 25 degrees C). Fluorescence intensity (Ex/Emmax = 394/509 nm) was measured immediately after the addition of GFP (8.0-9.0 microg/mL) into buffered or unbuffered chlorine solutions with or without constant stirring. With solutions constantly stirred, GFP fluorescence decreased abruptly on contact with chlorine in concentrations greater than 150 ppm, with D-values between 1.3 min (147 ppm chlorine) and 1.7 min (183 ppm chlorine). In phosphate buffered chlorine solutions (pH = 7.15 +/- 0.08), GFP retained its structure between 52 and 94 ppm, but protein stability decreased 10 fold when exposed to 110 ppm chlorine. The recovery of GFP fluorescence intensity due to renaturation was observed between 30 and 100 ppm chlorine in WFI (final pH = 11.01 +/- 0.23) without stirring. Stirring enhanced the contact between GFP and chlorine throughout the assay and provided a more accurate D-value evaluation. GFP performed as a suitable fluorescent marker for monitoring disinfection effectiveness. PMID- 17137322 TI - Molecular cloning and characterization of NAD(+)-dependent xylitol dehydrogenase from Candida tropicalis ATCC 20913. AB - Induction of xylitol dehydrogenase of Candida tropicalis ATCC 20913 by various carbon sources was investigated. The enzyme activity was induced when the yeast was grown on l-arabinose and d-xylose. A novel gene encoding the enzyme was cloned and characterized. The 1,095-bp coding sequence of the gene encodes a polypeptide of 364 amino acids, with a molecular mass of 39.4 kDa. Sequence analysis of the putative protein showed it to be a member of the zinc-containing alcohol dehydrogenase family and to have homology to xylitol dehydrogenase genes from other yeasts and fungi. The recombinant xylitol dehydrogenase expressed in Escherichia coli oxidized polyols such as xylitol and d-sorbitol and reduced ketoses such as d-xylulose and d-fructose. It required exclusively NAD or NADH as a cofactor. PMID- 17137323 TI - Diffusion of [3H]dexamethasone in rat subcutaneous slices after injection measured by digital autoradiography. AB - A relatively simple method for the determination of the diffusion coefficient of a substance that has been injected into tissue is described. We illustrate this method using [3]dexamethasone injected into the subcutaneous tissue of rats. Digital autoradiography was used to measure the distribution of the [3H] dexamethasone within the subcutaneous tissue at 2.5 and 20 min after injection. Measured concentration profiles of the injection were compared to a mathematical model of drug diffusion from an injection. There was good agreement between the experimental data and the mathematical model. The diffusion coefficient found using this simple injection method was (4.01 +/- 2.01) x 10(-10) m2/s. This D value was very close to the value of D = (4.11 +/- 1.77) x 10(-10) m2/s found previously using different mathematical and experimental techniques with osmotic pumps implanted for 6, 24, and 60 h in rats (1). The simple method given here for the determination of the diffusion coefficient is general enough to be applied to other substances and tissues as well. PMID- 17137324 TI - Screening extremophiles for bioconversion potentials. AB - A screening protocol was developed and implemented to evaluate extremophiles, in particular hyperthermophiles and thermoacidophiles, for their capacity to transform starch-based feedstocks to high-value organic acids and solvents. Screening results of 14 extremophiles showed promising growth and biotransformation potentials. In particular, Hyperthermus butylicus, Thermococcus litoralis, and Sulfolobus acidocaldarius were identified as producers of both organic acids and solvents under the screening protocol. The screening effort presented here represents an important step toward realization of biotransformation potentials of extremophiles, potentially improving upon biomass based processes. PMID- 17137325 TI - Quality assessment of tandem mass spectra based on cumulative intensity normalization. AB - A large proportion of MS/MS spectral analyses do not result in significant matches because their spectral quality is too poor to produce meaningful identification. Throughput of peptide identification can be greatly improved, if one can filter out, in advance, spectra that would lead to wrong identification. We introduce here an innovative approach to assess spectral quality utilizing a new spectral feature called Xrea, based on cumulative intensity normalization. PMID- 17137326 TI - Protein-RNA cross-linking in the ribosomes of yeast under oxidative stress. AB - Living systems have efficient degradative pathways for dealing with the fact that reactive oxygen species (ROS) derived from cellular metabolism and the environment oxidatively damage proteins and DNA. But aggregation and cross linking can occur as well, leading to a series of problems including disruption of cellular regulation, mutations, and even cell death. The mechanism(s) by which protein aggregation occurs and the macromolecular species involved are poorly understood. In the study reported here, evidence is provided for a new type of aggregate between proteins and RNA in ribosomes. While studying the effect of oxidative stress induced in the yeast proteome it was noted that ribosomal proteins were widely oxidized. Eighty six percent of the proteins in yeast ribosomes were found to be carbonylated after stressing yeast cell cultures with hydrogen peroxide. Moreover, many of these proteins appeared to be cross-linked based on their coelution patterns during RPC separation. Since they were not in direct contact, it was not clear how this could occur unless it was through the RNA separating them in the ribosome. This was confirmed in a multiple-step process, the first being derivatization of all carbonylated proteins in cell lysates with biotin hydrazide through Schiff base formation. Following reduction of Schiff bases with sodium cyanoborohydride, biotinylated proteins were selected from cell lysates with avidin affinity chromatography. Oxidized proteins thus captured were then selected again using boronate affinity chromatography to capture vicinal diol-containing proteins. This would include proteins cross linked to an RNA fragment containing a ribose residue with 2',3'-hydroxyl groups. Some glycoproteins would also be selected by this process. LC/MS/MS analyses of tryptic peptides derived from proteins captured by this process along with MASCOT searches resulted in the identification of 37 ribosomal proteins that appear to be cross-linked to RNA. Aggregation of proteins with ribosomal RNA has not been previously reported. The probable impact of this phenomenon cells is to diminish the protein synthesis capacity. PMID- 17137327 TI - Pharmacoproteomics study of cetuximab in nasopharyngeal carcinoma. AB - Epidermal growth factor receptor (EGFR) is usually overexpressed in nasopharyngeal carcinoma (NPC). Our recent in vitro study has demonstrated that cetuximab (an antibody drug against EGFR) inhibits the growth of NPC cell lines, HK1 and HONE-1. The present study investigates the effect of cetuximab on protein expressions of NPC cell lines. NPC cells were cultured in the absence or presence of cetuximab at the IC50 concentrations (3 nM for HK1 and 0.3 nM for HONE-1) for 48 h, and total cell lysates were extracted. The cell lysates were then subjected to two-dimensional polyacrylamide gel electrophoresis (2D PAGE), and the 2D gel images were compared to discover the protein changes caused by cetuximab treatment. The common differentially expressed proteins in NPC cell lines were identified by peptide mass fingerprinting. We found that heat shock protein gp96 was down-regulated, while alpha-enolase, tumor suppressor protein maspin, and p97 valosin containing protein were up-regulated after cetuximab treatment. Reverse transcription polymerase chain reaction (RT-PCR) analysis confirmed that the changes in protein levels of gp96, maspin, and p97 coincided with mRNA levels, indicating that these proteins were regulated at transcriptional levels. Up regulation of gp96 has been observed in various cancers and reported to have tumor protective effects. P97 is a multifunctional AAA (ATPase associated with a variety of activities) protein and is involved in numerous cellular activities including membrane transport, protein folding, protein degradation, and cell division. Maspin has been shown to increase apoptosis, and block the growth, invasion, and metastatic properties of many tumors. The comparative tumor suppression effects of cetuximab and maspin suggest that cetuximab might exert its antitumor effects partly by up-regulation of maspin expression. The study also indicates that proteomic analysis is a promising approach to elucidate the functional mechanisms of anticancer drugs. Pharmacoproteomic study may also help to identify clinical responders for drug treatment and provide insight for new drug development. PMID- 17137328 TI - Systematic identification of the subproteome of Escherichia coli cell envelope reveals the interaction network of membrane proteins and membrane-associated peripheral proteins. AB - Membrane proteins of Gram-negative bacteria are key molecules that interface the cells with the environment. Despite recent proteomic identification of numerous oligomer proteins in the Escherichia coli cell envelope, the protein complex of E. coli membrane proteins and their peripherally associated proteins remain ill defined. In the current study, we systematically analyze the subproteome of E. coli cell envelope enriched in sarcosine-insoluble fraction (SIF) and sarcosine soluble fraction (SSF) by using proteomic methodologies. One hundred and four proteins out of 184 spots on 2D electrophoresis gels are identified, which includes 31 outer membrane proteins (OMPs). Importantly, our further proteomic studies reveal a number of previously unrecognized membrane-interacting protein complexes, such as the complex consisting of OmpW and fumarate reductase. This established complete proteomic profile of E. coli envelope also sheds new insight into the function(s) of E. coli outer envelope. PMID- 17137329 TI - Mild protease treatment as a small-scale biochemical method for mitochondria purification and proteomic mapping of cytoplasm-exposed mitochondrial proteins. AB - Because of its importance in basic biology and medicine, great efforts are being devoted to unraveling of the genuine mitochondrial proteome, which is the dynamic protein complement that the organelle uses to maintain its structure and functionality. Several proteomic investigations have now clearly shown that all the purification approaches we have at our disposal suffer from the problem of co purification; therefore, it is very difficult to distinguish novel mitochondrial proteins from those that are just contaminants of the preparation. The question is further complicated by the fact that the mitochondrial proteome depends on the tissue source. Density gradient centrifugation is the most widespread purification method for obtaining highly pure mitochondrial fractions. The main disadvantage of these methods is the low yield of purified mitochondria that precludes their use in low-scale purifications. Here, we have treated small aliquots of crude mitochondria from mouse liver and from cultured hepatocytes (HEPA1-6) with trypsin under mild proteolysis conditions and have evaluated the suitability of this reaction as a small-scale purification approach. The protease removed several cytoplasmic and endoplasmic reticulum proteins, together with a fraction of mitochondrial proteins that we hypothesize to be associated with the cytosolic face of the outer mitochondrial membrane. The peculiar topology of these mitochondrial proteins could be indicative of their functional roles. Finally, our study represents an application of advanced mass spectrometry technology to the evaluation of biochemical approaches for the treatment of mitochondria. PMID- 17137330 TI - Rapid expression of functional genomic libraries. AB - Genomic-scale analysis of protein function is currently limited by the ability to rapidly express the enormous diversity of protein targets in their active form. We describe a method to construct transcriptionally active expression templates (ETs) in parallel using a single PCR step wherein the overlap-extension reaction for addition of transcription regulatory elements is separated from the amplification of the full-length product by using a GC-rich single primer. Over 90% of 55 diverse genomic targets were extended with T7 regulatory elements to form ETs in high yield and purity. The unpurified ETs directed protein expression using a cell-free protein synthesis (CFPS) system supplemented with cofactors and metal ions to activate a variety of enzymes. Higher activities were obtained in the modified CFPS reactions compared to standard reaction conditions. Protein purification was avoided because the expressed enzyme activity was significantly greater than the background activity associated with the cell extract. These improvements in the parallel synthesis of linear ETs combined with enhanced in vitro enzyme activation help to make CFPS systems more attractive platforms for high-throughput evaluation of protein function. PMID- 17137331 TI - The nuclear proteome of chickpea (Cicer arietinum L.) reveals predicted and unexpected proteins. AB - Nuclear proteins constitute a highly organized, complex network that plays diverse roles during cellular development and other physiological processes. The yeast nuclear proteome corresponds to about one-fourth of the total cellular proteins, suggesting the involvement of the nucleus in a number of diverse functions. In an attempt to understand the complexity of plant nuclear proteins, we have developed a proteome reference map of a legume, chickpea, using two dimensional gel electrophoresis (2-DE). Approximately, 600 protein spots were detected, and LC-ESI-MS/MS analyses led to the identification of 150 proteins that have been implicated in a variety of cellular functions. The largest percentage of the identified proteins was involved in signaling and gene regulation (36%), while 17% were involved in DNA replication and transcription. The chickpea nuclear proteome indicates the presence of few new nuclear proteins of unknown functions vis-a-vis many known resident proteins. To the best of our knowledge, this is the first report of a nuclear proteome of an unsequenced genome. PMID- 17137332 TI - Prediction of liquid chromatographic retention times of peptides generated by protease digestion of the Escherichia coli proteome using artificial neural networks. AB - We developed a computational method to predict the retention times of peptides in HPLC using artificial neural networks (ANN). We performed stepwise multiple linear regressions and selected for ANN input amino acids that significantly affected the LC retention time. Unlike conventional linear models, the trained ANN accurately predicted the retention time of peptides containing up to 50 amino acid residues. In 834 peptides, there was a strong correlation (R2 = 0.928) between measured and predicted retention times. We demonstrated the utility of our method by the prediction of the retention time of 121,273 peptides resulting from LysC-digestion of the Escherichia coli proteome. Our approach is useful for the proteome-wide characterization of peptides and the identification of unknown peptide peaks obtained in proteome analysis. PMID- 17137333 TI - Elevated levels of phosphorylated fibrinogen-alpha-isoforms and differential expression of other post-translationally modified proteins in the plasma of ovarian cancer patients. AB - We evaluated the differentially expressed proteins in the plasma of ovarian cancer (OVC) patients using 2-D SDS-polyacrylamide gel electrophoresis (SDS-PAGE) with post-translational modification (PTM) specific stains after the removal of six high-abundance proteins. The pooled plasma from patients with stage III or IV OVC was compared to a pooled postmenopausal age-matched control. Several proteins were identified as differentially expressed in the plasma of OVC patients. Among them, the phosphorylated fibrinogen-alpha-chain isoform (containing fibrinopeptide-A) was found to be up-regulated. Previously in our laboratory, phosphorylated fibrinopeptide-A was found to be up-regulated in the low molecular weight fraction of serum derived from OVC patients. We examined the levels of phosphorylated fibrinogen-alpha-chain in each patient that constituted the pooled plasma using Western blot, mass spectrometry (MS), and PTM specific stains. Phosphoprotein bands containing fibrinogen-alpha-chain fragments showed up regulation in all OVC patients. PMID- 17137334 TI - Proteomic identification of altered proteins in skeletal muscle during chronic potassium depletion: Implications for hypokalemic myopathy. AB - Prolonged potassium depletion is a well-known cause of myopathy. The pathophysiology of hypokalemic myopathy, however, remains unclear. We performed a gel-based, differential proteomics study to define altered proteins in skeletal muscles during chronic potassium depletion. BALB/c mice were fed with normal chow (0.36% K+) or K+-depleted (KD) diet (<0.001% K+) for 8 weeks (n = 5 in each group). Left gastrocnemius muscles were surgically removed from each animal. Histopathological examination showed mild-degree infiltration of polymornuclear and mononuclear cells at the interstitium of the KD muscles. Extracted proteins were resolved with two-dimensional electrophoresis (2-DE), and visualized with Coomassie Brilliant Blue R-250 stain. Quantitative intensity analysis revealed 16 up-regulated protein spots in the KD muscles, as compared to the controls. These differentially expressed proteins were subsequently identified by peptide mass fingerprinting and by quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS/MS). Most of the altered proteins induced by chronic potassium depletion were muscle enzymes that play significant roles in several various metabolic pathways. Other up-regulated proteins included myosin-binding protein H, alpha-B Crystallin, and translationally controlled tumor protein (TCTP). These findings may lead to a new roadmap for research on hypokalemic myopathy, to better understanding of the pathophysiology of this medical disease, and to biomarker discovery. PMID- 17137335 TI - VDAC1, having a shorter N-terminus than VDAC2 but showing the same migration in an SDS-polyacrylamide gel, is the predominant form expressed in mitochondria of various tissues. AB - The voltage-dependent anion channel (VDAC) is a pore-forming protein expressed in the outer membrane of eukaryotic mitochondria. Three isoforms of it, i.e., VDAC1, VDAC2, and VDAC3, are known to be expressed in mammals; however, the question as to which is the main isoform in mitochondria is still unanswered. To address this question, we first prepared standard VDACs by using a bacterial expression system and raised various antibodies against them by using synthetic peptides as immunogens. Of the three bacterially expressed VDAC isoforms, VDAC3 showed faster migration in SDS-polyacrylamide gels than VDAC1 and VDAC2, although VDAC2 is longer than VDAC1 and VDAC3, due to a 12-amino acid extension of its N-terminal region. Even with careful structural characterization of the expressed VDACs by LC-MS/MS analysis, serious structural modifications of VDACs causing changes in their migration in SDS-polyacrylamide gels were not detected. Next, immunoreactivities of the raised antibodies toward these bacterially expressed VDAC isoforms were evaluated. Trials to prepare specific antibodies against the three individual VDAC isoforms were not successful except in the case of VDAC1. However, using a synthetic peptide corresponding to the highly conserved region among the three VDACs, we were successful in preparing an antibody showing essentially equal immunoreactivities toward all three VDACs. When mitochondrial outer membrane proteins of various rat tissues were subjected to 2-dimensional electrophoresis followed by immunoblotting with this antibody, six immunoreactive protein spots were detected. These spots were characterized by LC-MS/MS analysis, and the signal intensities among the spots were compared. As a result, the signal intensity of the spot representing VDAC1 was the highest, and thus, VDAC1 was concluded to be the most abundantly expressed of the three VDAC isoforms in mammalian mitochondria. PMID- 17137336 TI - Characterization of the human pancreatic islet proteome by two-dimensional LC/MS/MS. AB - The pancreatic beta-cell plays a central role in the maintenance of glucose homeostasis and in the pathogenesis of both type 1 and type 2 diabetes mellitus. Elucidation of the insulin secretory defects observed in diabetes first requires a better understanding of the complex mechanisms regulating insulin secretion, which are only partly understood. While there have been reports detailing proteomic analyses of islet cell lines or isolated rodent islets, the information gained is not always applicable to humans. Therefore, definition of the human islet proteome could contribute to a better understanding of islet biology and lead to more effective treatment strategies. We have applied a two-dimensional LC MS/MS-based analysis to the characterization of the human islet proteome, resulting in the confident identification of 29,021 different tryptic peptides covering 3365 proteins (> or =2 unique peptide identifications per protein). As expected, the three major islet hormones (insulin, glucagon, and somatostatin) were detected, as well as various beta-cell enriched secretory products, ion channels, and transcription factors. In addition, significant proteome coverage of metabolic enzymes and cellular pathways was observed, including the integrin signaling cascade and the MAP kinase, NF-kappa beta, and JAK/STAT pathways. The resulting peptide reference library provides a resource for future higher throughput and quantitative studies of islet biology. PMID- 17137337 TI - The Medicago truncatula small protein proteome and peptidome. AB - The small protein and native peptide component of plant tissues is a neglected area of proteomic studies. We have used fractionation techniques for denatured and nondenatured protein preparations combined with 2-D LC tandem mass spectrometry to examine the sequences of small proteins and peptides in four tissues of the model legume, Medicago truncatula: the root tip and root of germinating seedlings, nitrogen fixing nodules, and young leaves. The isolation and fractionation strategies successfully enriched the small protein and native peptide content of the samples. Eighty-one small M. truncatula proteins and native peptides were identified. Most samples were dominated by ribosomal and histone proteins, and leaf samples possessed photosynthesis-related proteins. Secreted proteins such as lipid transfer proteins were common to several tissues. Twenty-four hours after germination, the roots and root tip tissues possessed several "seed-specific" and late-embryogenesis proteins. We conclude that these proteins are present in cells prior to germination and that they are subsequently used as a nutritional source for the young tissues. Native UV absorbing peptides were detected in very low molecular weight fractions and sequenced. Each peptide shared C-terminal residues and showed homology to the seed storage protein legumin. The strategies used here would be suitable for combining bioassays and mass spectrometry to identify bioactive peptides in the M. truncatula peptidome. PMID- 17137338 TI - Rat neuropeptidomics by LC-MS/MS and MALDI-FTMS: Enhanced dissection and extraction techniques coupled with 2D RP-RP HPLC. AB - Recently developed sample preparation techniques employing microwave irradiation have enabled the comprehensive study of endogenous mammalian neuropeptides. These methods reduce interference from post-mortem protein degradation by deactivating proteases via heat denaturation. Alternatively, we have developed a protocol using cryostat dissection and a boiling extraction buffer to achieve a similar effect. This novel methodology greatly reduces post-mortem protein contamination and increases neuropeptide identification without the use of specialized equipment. In addition, a 2D HPLC scheme employing differential pH selectivity in the first and second dimensions has been used to enhance neuropeptidome coverage. By using our novel dissection protocol in tandem with 2D RP-RP HPLC, we were able to identify a total of 56 peptides from known neuropeptide precursors, including 17 previously unidentified peptides. The use of cryostat dissection and two dimensional RP-RP HPLC enhances the detection of novel neuropeptides by deactivating proteases and reducing sample complexity. PMID- 17137339 TI - Glycoproteomics of Trypanosoma cruzi trypomastigotes using subcellular fractionation, lectin affinity, and stable isotope labeling. AB - Herein we detail the first glycoproteomic analysis of a human pathogen. We describe an approach that enables the identification of organelle and cell surface N-linked glycoproteins from Trypanosoma cruzi, the causative agent of Chagas' disease. This approach is based on a subcellular fractionation protocol to produce fractions enriched in either organelle or plasma membrane/cytoplasmic proteins. Through lectin affinity capture of the glycopeptides from each subcellular fraction and stable isotope labeling of the glycan attachment sites with H(2)18O, we unambiguously identified 36 glycosylation sites on 35 glycopeptides which mapped to 29 glycoproteins. We also present the first expression evidence for 11 T. cruzi specific glycoproteins and provide experimental data indicating that the mucin associated surface protein family (MASP) and dispersed gene family (DGF-1) are post-translationally modified by N linked glycans. PMID- 17137340 TI - Global comparison of the membrane subproteomes between a multidrug-resistant Acinetobacter baumannii strain and a reference strain. AB - Acinetobacter baumannii causes severe infections in compromised patients. We combined SDS-PAGE, two-dimensional gel electrophoresis and mass spectrometry (LC MS/MS and MALDI-TOF) to separate and characterize the proteins of the cell envelope of this bacterium. In total, 135 proteins (inner and outer membrane proteins) were identified. In this analysis, we described the expression by this bacterium of RND-type efflux systems and some potential virulence factors. We then compared the membrane subproteome of a clinical multidrug-resistant (MDR) isolate with that of a reference strain. We found that the MDR strain expressed lower levels of the penicillin-binding-protein 1b, produced a CarO protein having different primary and quaternary structures to that of the reference strain, and expressed OmpW isoforms. We also showed that the clinical strain has a high ability to form biofilms consistent with the accumulation of some outer membrane proteins (OMPs) such as NlpE or CsuD that have already been described as involved in bacterial adhesion. These features may partly explain the MDR emergence of the clinical isolate. PMID- 17137341 TI - Challenges related to analysis of protein spot volumes from two-dimensional gel electrophoresis as revealed by replicate gels. AB - Assumptions that need to be considered prior to statistical analysis of protein spot volumes from two-dimensional gel electrophoresis (2-DE) data are studied using replicate gels of the same sample. The most important observation is that the data tables of protein spot volumes from 2-DE images contain a large number of missing values, which are not consistent with the presence or absence of the proteins. This implies both loss of information and problems for the subsequent statistical analysis. Challenges with 2-DE protein spot volumes are viewed in light of multiple gel comparisons and multivariate data analysis. PMID- 17137342 TI - Proteomic analysis of Saccharomyces cerevisiae under high gravity fermentation conditions. AB - Saccharomyces cerevisiae KAY446 was utilized for ethanol production, with glucose concentrations ranging from 120 g/L (normal) to 300 g/L (high). Although grown in a high glucose environment, S. cerevisiae still retained the ability to produce ethanol with a high degree of glucose utilization. iTRAQ-mediated shotgun proteomics was applied to identify relative expression change of proteins under the different glucose conditions. A total of 413 proteins were identified from three replicate, independent LC-MS/MS runs. Unsurprisingly, many proteins in the glycolysis/gluconeogenesis pathway showed significant changes in expression level. Twenty five proteins involved in amino acid metabolism decreased their expression, while the expressions of 12 heat-shock related proteins were also identified. Under high glucose conditions, ethanol was produced as a major product. However, the assimilation of glucose as well as a number of byproducts was also enhanced. Therefore, to optimize the ethanol production under very high gravity conditions, a number of pathways will need to be deactivated, while still maintaining the correct cellular redox or osmotic state. Proteomics is demonstrated here as a tool to aid in this forward metabolic engineering. PMID- 17137343 TI - Large-scale predictions of gram-negative bacterial protein subcellular locations. AB - Many species of Gram-negative bacteria are pathogenic bacteria that can cause disease in a host organism. This pathogenic capability is usually associated with certain components in Gram-negative cells. Therefore, developing an automated method for fast and reliable prediction of Gram-negative protein subcellular location will allow us to not only timely annotate gene products, but also screen candidates for drug discovery. However, protein subcellular location prediction is a very difficult problem, particularly when more location sites need to be involved and when unknown query proteins do not have significant homology to proteins of known subcellular locations. PSORT-B, a recently updated version of PSORT, widely used for predicting Gram-negative protein subcellular location, only covers five location sites. Also, the data set used to train PSORT-B contains many proteins with high degrees of sequence identity in a same location group and, hence, may bear a strong homology bias. To overcome these problems, a new predictor, called "Gneg-PLoc", is developed. Featured by fusing many basic classifiers each being trained with a stringent data set containing proteins with strictly less than 25% sequence identity to one another in a same location group, the new predictor can cover eight subcellular locations; that is, cytoplasm, extracellular space, fimbrium, flagellum, inner membrane, nucleoid, outer membrane, and periplasm. In comparison with PSORT-B, the new predictor not only covers more subcellular locations, but also yields remarkably higher success rates. Gneg-PLoc is available as a Web server at http://202.120.37.186/bioinf/Gneg. To support the demand of people working in the relevant areas, a downloadable file is provided at the same Web site to list the results identified by Gneg-PLoc for 49 907 Gram-negative protein entries in the Swiss-Prot database that have no subcellular location annotations or are annotated with uncertain terms. The large-scale results will be updated twice a year to cover the new entries of Gram-negative bacterial proteins and reflect the new development of Gneg-PLoc. PMID- 17137344 TI - PIGOK: Linking protein identity to gene ontology and function. AB - Here we introduce a computer database that allows for the rapid retrieval of physicochemical properties, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes information about a protein or a list of proteins. We applied PIGOK analyzing Schizosaccharomyces pombe proteins displaying differential expression under oxidative stress and identified their biological functions and pathways. The database is available on the Internet at http://pc4 133.ludwig.ucl.ac.uk/pigok.html. PMID- 17137345 TI - Microfluidic protein detection through genetically engineered bacterial cells. AB - Protein microarray technology, in which a large number of capture ligands are spatially arrayed at a high density, presents an attractive method for high throughput proteomic analysis. Toward this end, we demonstrate the first cell based protein detection in a microsystem, wherein Escherichia coli cells are genetically engineered to express the desired capture proteins on the membrane surface and are spatially arrayed as sensing elements in a microfluidic device. An E. coli clone expressing peptide ligands with high affinity and high specificity for target molecules was isolated a priori. Then these cells were electrokinetically immobilized on gold electrodes using dielectrophoresis, thus allowing each sensor element to be electrically addressable. Flow cytometry and subsequent fluorescence analysis verified the highly specific capture and detection of target molecules by the bacteria. Finally, through the coexpression of peptide-based capture ligands on the cell surface and fluorescent protein in the cytoplasm, we demonstrate an effective means of directly linking the fluorescence intensity to the density of capture ligands. PMID- 17137346 TI - A new functional, chemical proteomics technology to identify purine nucleotide binding sites in complex proteomes. AB - Adenine nucleotides are small, abundant molecules that bind numerous proteins involved in pivotal cellular processes. These nucleotides are co-factors or substrates for enzymes, regulators of protein function, or structural binding motifs. The identification of nucleotide-binding sites on a proteome-wide scale is tempting in view of the high number of nucleotide-binding proteins, their large in vivo concentration differences, and the various functions they exert. Here, we report on a functional, chemical, gel-free proteomics technology that allows the identification of protein adenine nucleotide-binding site(s) in cell lysates. Our technology uses a synthetic ATP analogue, 5'-p fluorosulfonylbenzoyladenosine (FSBA), as an affinity/activity-based probe for nucleotide-binding sites. When applied on a cellular level, 185 different FSBA labeled sites in a human Jurkat cell lysate were identified. Functional and structural aspects of the use of FSBA on a proteome-wide scale are discussed. PMID- 17137347 TI - A clean, more efficient method for in-solution digestion of protein mixtures without detergent or urea. AB - Proteolytic digestion of a complicated protein mixture from an organelle or whole cell lysate is usually carried out in a dilute solution of a denaturing buffer, such as 1-2 M urea. Urea must be subsequently removed by C18 beads before downstream analysis such as HPLC/MS/MS or complete methylation followed by IMAC isolation of phosphopeptides. Here we describe a procedure for digesting a complicated protein mixture in the absence of denaturants. Proteins in the mixture are precipitated with trichloroacetic acid/acetone for denaturation and salt removal and resuspended in NH4HCO3 buffer. After trypsinolysis, the resulting peptides are not contaminated by urea or other nonvolatile salts and can be dried in a SpeedVac to remove NH4HCO3. When this protocol was applied to an extract of A431 cells, 96.8% of the tryptic peptides were completely digested (i.e., had no missed cleavage sites), in contrast to 87.3% of those produced by digestion in urea buffer. We successfully applied this digestion method to analysis of the phosphoproteome of adiposomes from HeLa cells, identifying 33 phosphorylation sites in 28 different proteins. Our digestion method avoids the need to remove urea before HPLC/MS/MS analysis or methylation and IMAC, increasing throughput while reducing sample loss and contamination from sample handling. We believe that this method should be valuable for proteomics studies. PMID- 17137348 TI - Hexadecanedionic acid-sepharose 4B: A new tool for preparation of albumin depleted plasma. AB - Serum and plasma are the major sources of human material for clinical molecular diagnostics and drug discovery. However, due to the high abundance of some proteins, of which serum albumin (SA) is most prominent, lower-abundance proteins often remain undetectable in proteomic analysis of these body fluids. We have used hexadecanedionic acid (HDA) immobilized to Sepharose 4B to develop an affinity resin that is effective in the removal of SA from plasma. Two dimensional gel analysis of the SA-depleted samples shows a significant enhancement of the low-abundance proteins and highly specific capture of serum albumin. The HDA resin shows better performance in terms of specificity than dye based resins. PMID- 17137349 TI - Analysis of the soluble ATP-binding proteome of plant mitochondria identifies new proteins and nucleotide triphosphate interactions within the matrix. AB - The interactions of ATP inside plant mitochondria were investigated by identifying the soluble nucleotide binding proteome captured using immobilized ATP. Selected proteins were separated by 1D SDS-PAGE and 2D IEF-SDS-PAGE and identified by ESI-Q-TOF MS/MS. A range of highly enriched proteins were identified from the mitochondrial proteome, including 14-3-3 proteins and RNA binding proteins, as well as proteins known to contain nucleotide binding domains and/or to be inhibited or stimulated by ATP. PMID- 17137350 TI - Perspective on dirhodium carboxamidates as catalysts. AB - Dirhodium compounds are emerging as highly efficient catalysts for diverse reactions, and those with carboxamidate ligands have the broadest applications. The unique features of these compounds are their structural rigidity, ease of ligand exchange, open diaxial sites for coordination with Lewis bases, and their low oxidation potential. As consequences of this, dirhodium carboxamidates are efficient and effective catalysts for metal carbene reactions, Lewis acid catalyzed processes, and chemical oxidations. With chiral carboxamidate ligands these dirhodium compounds show exceptional enantiocontrol for intramolecular cyclopropanation and carbon-hydrogen insertion reactions of diazoacetates, and they are also highly efficient and selective for hetero-Diels-Alder reactions. Their limitations lie in their moderate reactivities for metal carbene generation and Lewis acid catalysis and in the cost of the precious metal rhodium. PMID- 17137351 TI - Substituent effects in C6F6-C6H5X stacking interactions. AB - Parallel displaced and sandwich configurations of hexafluorobenzene-substituted benzene dimers are studied by ab initio molecular orbital methods up to the MP2(full)/aug-cc-pVDZ level of theory to reveal how substituents influence pi-pi interactions. Two minimum energy configurations are found, one with the substituent group away from the pi-face of the hexafluorobenzene ring (2a-f) and the other with the substituent group on top of the pi-face of the hexafluorobenzene (3a-f). Higher binding energies are predicted for dimers with the substituent on the pi-face (3a-f). All sandwich dimers (4a-e) are found to be saddle points on the potential energy surfaces. A parallel-displaced minimum energy configuration is also predicted for the parent complex, C6F6-C6H6, which is in contrast to predictions based on quadrupole moments of benzene and hexafuorobenzene. The preference for the parallel displaced, rather than the sandwich configuration, is rationalized based on the smaller interplanar distance in the former. The closeness of contact in the parallel-displaced dimers leads to greater binding energies. The shape of the electron density isosurface of the monomers is suggested to provide a guide for the prediction of how arenes stack with one another. A large difference in binding energy between the C6F6 complex of aniline (3e) and N,N-dimethylaniline (3f) is calculated, and charge-transfer interactions are suggested to play a role in the latter. PMID- 17137352 TI - Aromaticity and Mobius antiaromaticity in monocyclic [11]annulenium cations. AB - Monocyclic [11]annulenium cations, which are experimentally unknown, have been studied primarily via DFT methods but also with some CCSD(T) validation. We have located six minima: two doubly trans (26, 27), one triply trans (28), one singly trans (29), one quintuply trans (trannulene-type, 33), and one all-cis (31). The first three are aromatic, 33 is modestly aromatic, 29 is nonaromatic, and the last is a Mobius antiaromatic species. We also investigated the fusion of various numbers of three-membered rings (3MRs) to the central 11-membered ring (11MR). We found several planar, all-cis-[11]annulenium ion derivatives as well as another Mobius antiaromatic species (52b); for comparison, we also found planar, antiaromatic all-cis-[12]annulene (60) and [15]annulenium cation (61) derivatives. The (anti)aromatic characterization of these compounds is based mainly on calculated magnetic data for the ground singlet and vertical triplet states, although aromatic stabilization energies (ASE) are also considered. Data for optimized triplets, several of which are Mobius aromatic systems (31t, 52t, 63t, 64t), are also included. Several of these cations are reasonable synthetic targets. PMID- 17137353 TI - New synthetic routes toward enantiopure nitrogen donor ligands. AB - New polypyridylic chiral ligands, having either C3 or lower symmetry, have been prepared via a de novo construction of the pyridine nucleus by means of Krohnke methodology in the key step. The chiral moieties of these ligands originate from the monoterpen chiral pool, namely (-)-alpha-pinene ((-)-14, (-)-15) and (-) myrtenal ((-)-9, (-)-10). Extension of the above-mentioned asymmetric synthesis procedure to the preparation of enantiopure derivatives of some commonly used polypyridylic ligands has been achieved through a new aldehyde building block (( )-16). As an example, the synthesis of a chiral derivative of N,N-bis(2 pyridylmethyl)ethylamine (bpea) ligand, (-)-19, has been performed to illustrate the viability of the method. The coordinative ability of the ligands has been tested through the synthesis and characterization of complexes [Mn((-)-19)Br2], ( )-20, and [RuCl((-)-10)(bpy)](BF4), (-)-21. Some preliminary results related to the enantioselective catalytic epoxidation of styrene with the ruthenium complex are also presented. PMID- 17137354 TI - An efficient and practical system for the catalytic oxidation of alcohols, aldehydes, and alpha,beta-unsaturated carboxylic acids. AB - Upon exposure to commercial bleach (approximately 5% aqueous sodium hypochlorite), nickel(II) chloride or nickel(II) acetate is transformed quantitatively into an insoluble nickel species, nickel oxide hydroxide. This material consists of high surface area nanoparticles (ca. 4 nm) and is a useful heterogeneous catalyst for the oxidation of many organic compounds. The oxidation of primary alcohols to carboxylic acids, secondary alcohols to ketones, aldehydes to carboxylic acids, and alpha, beta-unsaturated carboxylic acids to epoxy acids is demonstrated using 2.5 mol % of nickel catalyst and commercial bleach as the terminal oxidant. We demonstrate the controlled and selective oxidation of several organic substrates using this system affording 70-95% isolated yields and 90-100% purity. In most cases, the oxidations can be performed without an organic solvent, making this approach attractive as a "greener" alternative to conventional oxidations. PMID- 17137355 TI - Unexpected stereodynamic consequences of the restricted rotations in ortho-acyl- and ortho-vinyl biphenyls. AB - Surprisingly, the aryl-aryl rotation barriers of biphenyl derivatives ortho substituted by the "small" HC=O and HC=CH2 groups (10.0 and 8.4 kcal mol-1, respectively) were found greater than those observed in biphenyls ortho substituted by the "large" t-BuC=O and t-BuC=CH2 groups (6.7 and 6.9 kcal mol-1, respectively). PMID- 17137356 TI - Homolytic dissociation of the vulcanization accelerator tetramethylthiuram disulfide (TMTD) and structures and stabilities of the related radicals Me2NCSn* (n = 1-4). AB - The homolytic dissociation of the important vulcanization accelerator tetramethylthiuram disulfide (TMTD) has been studied by ab initio calculations according to the G3X(MP2) and G3X(MP2)-RAD theories. Homolytic cleavage of the SS bond requires a low enthalpy of 150.0 kJ mol-1, whereas 268.0 kJ mol-1 is needed for the dissociation of one of the C-S single bonds. To cleave one of the SS bonds of the corresponding trisulfide (TMTT) requires 191.1 kJ mol-1. Me2NCS2* is a particularly stable sulfur radical as reflected in the low S-H bond dissociation enthalpy of the corresponding acid Me2NC(=S)SH (301.7 kJ mol-1). Me2NCS2* (2B2) is a sigma radical characterized by the unpaired spin density shared equally between the two sulfur atoms and by a 4-center (NCS2) delocalized pi system. The ESR g-tensors of the radicals Me2NCSn* (n = 1-3) have been calculated. Both TMTD and the mentioned radicals form stable chelate complexes with a Li+ cation, which here serves as a model for the zinc ions used in accelerated rubber vulcanization. Although the binding energy of the complex [Li(TMTD)]+ is larger than that of the isomeric species [Li(S2CNMe2)2]+ (12), the dissociation enthalpy of TMTD as a ligand is smaller (125.5 kJ mol-1) than that of free TMTD. In other words, the homolytic dissociation of the SS bonds of TMTD is facilitated by the presence of Li+ ions. The sulfurization of TMTD in the presence of Li+ to give the paramagnetic complex [Li(S3CNMe2)2]+ is strongly exothermic. These results suggest that TMTD reacts with naked zinc ions as well as with the surface atoms of solid zinc oxide particles in an analogous manner producing highly reactive complexes, which probably initiate the crosslinking process during vulcanization reactions of natural or synthetic rubber accelerated by TMTD/ZnO. PMID- 17137357 TI - Enantiomerically pure isophorone diamine [3-(aminomethyl)-3,5,5 trimethylcyclohexylamine]: a chiral 1,4-diamine building block made available on large scale. AB - Isophorone diamine [IPDA, 3-(aminomethyl)-3,5,5-trimethylcyclohexylamine] is a chiral non-C2-symmetric 1,4-diamine which is produced industrially on large scale as the mixture of all four stereoisomers (cis/trans ca. 3:1). Starting from this industrial bulk product, the preparation of the bis-tosyl, bis-Fmoc, bis-Boc and bis-Z derivatives of cis-IPDA, the preparation of the pure cis enantiomers by HPLC on chiral stationary phase, and the assignment of absolute configurations to the isolated enantiomers are described. We furthermore report an efficient method for the optical resolution of IPDA by salt formation with dibenzoyl tartaric acid. The latter method conveniently affords enantiomerically pure cis-IPDA in 100 g quantities. A number of salen ligands have been prepared from this enantiomerically pure 1,4-diamine and fully characterized. The nickel complex of one of the salen ligands was prepared and analyzed by X-ray crystallography. The crystal structure of the Ni4L4 complex illustrates the pronounced preference of cis-IPDA for adopting the chair conformation in which both the amino- and the aminomethyl substituents occupy equatorial positions. As a consequence, the two salicylidene imine moieties of one ligand molecule do not converge on one metal ion, but act as bridging ligands between two nickel ions. PMID- 17137358 TI - 1,3-Dipolar cycloadditions of electrophilically activated benzonitrile N-oxides. Polar cycloaddition versus oxime formation. AB - The reactions of electrophilically activated benzonitrile N-oxides (BNOs) toward 3-methylenephthalimidines (MPIs) have been studied using density functional theory (DFT) at the B3LYP/6-31G* level. For these reactions, two different channels allowing the formation of the [3 + 2] cycloadducts and two isomeric (E)- and (Z)-oximes have been characterized. The 1,3-dipolar cycloadditions take place along concerted but highly asynchronous transition states, while formation of the oximes is achieved through a stepwise mechanism involving zwitterionic intermediates. Both reactions are initiated by the nucleophilic attack of the methylene carbon of the MPIs to the carbon atom of the electrophilically activated BNOs. The analysis based on the natural bond orbital (NBO) and the topological analysis of the electron localization function (ELF) at the transition structures and intermediates explains correctly the polar nature of these reactions. Solvent effects considered by the PCM model allow explaining the low incidence of the solvent polarity on the rate and composition of the reactions. PMID- 17137359 TI - Fries rearrangement of aryl formates: a mechanistic study by means of 1H, 2H, and 11B NMR spectroscopy and DFT calculations. AB - 1H, 2H, and 11B NMR spectroscopy has been used to study the mechanism of the Fries rearrangement of aryl formates promoted by boron trichloride by monitoring both the substrate and the Lewis acid. DFT calculations were employed to investigate the energetics of several reaction paths and to calculate NMR chemical shifts of key intermediates and products. After the formation of a 1:1 substrate-Lewis acid adduct, the rearrangement proceeds in two steps, beginning with the cleavage of the ester bond and the release of formyl chloride in situ, which, in turn, acts as a formylating agent, introducing an aldehydic functionality into the aromatic ring. The high regioselectivity (only the ortho product is obtained) is also accounted for by the proposed intermolecular, Lewis acid-assisted mechanism. PMID- 17137360 TI - Crystallography and magnetism of two 1-(4-nitroxylphenyl)pyrroles. AB - Stable radicals 1-(4-(N-tert-butyl-N-aminoxyl)phenyl)pyrrole (BNPP) and 1-(4-(N [para-methoxyphenyl]-N-aminoxyl)phenyl)pyrrole (MNPP) were synthesized and characterized by crystallography and magnetism. BNPP crystals exhibit 1-D chains of intermolecular nitroxide NO to nitroxide CH3 contacts, but polycrystalline magnetic susceptibility measurements show quite small antiferromagnetic (AFM) exchange interactions. MNPP shows stronger AFM exchange interactions that appear to be associated with a 2-D planar mesh of crystallographic nitroxide to nitroxide (N)O...N(O) contacts of 4.0-4.2 A. The AFM behavior of MNPP can be fitted to a 2-D square planar Heisenberg antiferromagnetic exchange model with J/k = (-)0.78 +/- 0.04 K and mean field constant theta = (-)0.77 +/- 0.12 K. PMID- 17137361 TI - Ancistrocladinium A and B, the first N,C-coupled naphthyldihydroisoquinoline alkaloids, from a Congolese ancistrocladus species. AB - The isolation and structural elucidation of three novel-type naphthylisoquinoline alkaloids, ancistrocladinium A and B (the latter along with its atropisomer), from a Congolese Ancistrocladus species collected in the habitat Yeteto is reported. Their structures, including all stereochemical features, were elucidated by spectroscopic, chemical, and chiroptical methods. Ancistrocladinium A and B are the first N,C-coupled naphthyldihydroisoquinoline alkaloids found in nature, i.e., with an iminium-aryl axis. Although ancistrocladinium A, which is N,8'-coupled, is configurationally stable at this axis, ancistrocladinum B and its rotational isomer are based on a hitherto unprecedented N,6'-coupling type, with a slow rotation about the hetero biaryl axis at room temperature; they thus occur as a 46:54 mixture of two configurationally semistable atropo diastereomers. For the isomerization of (P)-ancistrocladinium B to its (M) diastereomer and for the opposite direction, the Gibbs free energies of activation were determined to be DeltaG double dagger1 = 105.8 kJ mol-1 and DeltaG double dagger2 = 105.7 kJ mol-1, respectively. In addition, the compounds were shown to have promising antileishmanial activities. PMID- 17137362 TI - The question of aromaticity in open-shell cations and anions as ion-radical offsprings of polycyclic aromatic and antiaromatic hydrocarbons. AB - Arene cation-radicals and anion-radicals result directly from the one-electron oxidation and reduction of many aromatic hydrocarbons, yet virtually nothing is known of their intrinsic (thermodynamic) stability and hence "aromatic character". Since such paramagnetic ion radicals lie intermediate between aromatic (Huckel) hydrocarbons with 4n + 2-electrons and antiaromatic analogues with 4n-electrons, we can now address the question of pi-delocalization in these odd-electron counterparts. Application of the structure-based "harmonic oscillator model of aromaticity" or the HOMA method leads to the surprising conclusion that the aromaticity of these rather reactive, kinetically unstable arene cation and anion radicals (as measured by the HOMA index) is actually higher than that of their (diamagnetic) parent-contrary to conventional expectations. PMID- 17137363 TI - Au(I)-catalyzed cyclization of enynes bearing an olefinic cycle. AB - Gold(I)-catalyzed cyclization of enynes containing an olefinic cycle has been studied. The introduction of an olefinic ring instead of a terminal alkene in enynes dramatically increased the yield of the reaction. Enynes having an olefinic cycle were prepared by a rhodium-catalyzed intermolecular [4 + 2] cycloaddition of diynes with butadiene. Consecutive rhodium-catalyzed Diels Alder/gold(I)-catalyzed cycloisomerization reactions were integrated in a one-pot reaction. PMID- 17137364 TI - Asymmetric synthesis of 2-(2-pyridyl)aziridines from 2-pyridineimines bearing stereogenic N-alkyl substituents and regioselective opening of the aziridine ring. AB - The addition of chloromethyllithium to the imine derived from 2 pyridinecarboxaldehyde and (S)-valinol, protected as its O-trimethylsilyl ether, gave the 1,2-disubstituted aziridine with good yield and diastereoselectivity. The analogous reaction performed on the imine derived from (S)-valine methyl ester gave the product containing the aziridine ring and the alpha-chloro ketone group coming from the attack of chloromethyllithium to the ester function. Other stereogenic alkyl substituents at nitrogen gave less satisfactory results. Moreover, the aziridination protocol did not work on other aromatic imines which were not capable of bidentate chelation, e.g., 3- and 4-pyridineimine and benzaldimine. Preliminary studies showed the possibility to carry out regio- and stereospecific opening reactions of 2-(2-pyridyl)aziridines by attack of internally generated or external nucleophiles. PMID- 17137365 TI - Photochemistry of o-pyrrolylstilbenes and formation of spiro-2H-pyrroles and their rearrangement to dihydroindoles. AB - Excited states of stilbenylpyrroles 1a-1c deactivate by two photochemical processes: cis-trans-isomerization and hydrogen transfer of NH to the stilbene double bond. NH-transfer results in the formation of two quinone dimethane intermediates, 10 and 11, and biradicals 12. Intramolecular cyclization of intermediates 10-12 gives rise to polycyclic compounds spiro-2H-pyrroles 7, pyrroloisoindoles 3, and pyrroloisoquinolines 8. Spiro-2H-pyrroles 7 rearrange on silica gel, giving dihydroindoles 2. PMID- 17137366 TI - Synthetic studies toward (-)-FR901483 using a conjugate allylation to install the C-1 quaternary carbon. AB - Two approaches to the aza-tricyclo dodecane skeleton of (-)-FR901483 are reported. Both routes utilized a Grignard addition to an N-acylpyridinium salt to establish the absolute stereochemistry at C-6 and a highly diastereoselective conjugate allylation reaction to form the quaternary center at C-1 of the natural product in an excellent yield. Although the desired polysubstituted piperidine intermediates were prepared regio- and stereoselectively, the construction of the C-8/C-9 bond connectivity could not be achieved. All attempts at a pinacol cyclization or an intramolecular 6-exo-tet epoxide opening were unsuccessful because of an unfavorable A(1,3) strain inherent in the molecule. PMID- 17137367 TI - N-H carbazole synthesis from 2-chloroanilines via consecutive amination and C-H activation. AB - N-H carbazoles can be produced from 2-chloroanilines and aryl bromides via consecutive catalytic amination and C-H activation. In many instances, this can be done in a tandem manner in one pot. The methodologies developed can be used in the synthesis of a range of carbazoles, including the natural products Clausine P and glycozolidine and a precursor in the synthesis of Clausines H, K, O, and 7 methoxy-O-methylmukonal, and can be extended to the synthesis of indoles. PMID- 17137368 TI - Ring closing metathesis/fragmentation route to (Z)-configured medium ring cycloalkenes. Total synthesis of (+/-)-periplanone C. AB - The combination of ring closing metathesis and beta-fragmentation offers an efficient entry into (Z)-configured medium ring cycloalkenes. The fragmentation step can be effected under anionic or radical conditions. The versatility of this method is demonstrated by the total synthesis of (+/-)-periplanone C-a macrocyclic pheromone of Periplaneta americana. PMID- 17137369 TI - Synthesis of methylene- and difluoromethylenephosphonate analogues of uridine-4 phosphate and 3-deazauridine-4-phosphate. AB - Cytidine triphosphate synthetase (CTPS) catalyzes the formation of cytidine triphosphate from glutamine, uridine-5'-triphosphate (UTP), and adenosine-5' triphosphate. Inhibitors of CTPS are of interest because of their potential as therapeutic agents. One approach to potent enzyme inhibitors is to use analogues of high energy intermediates formed during the reaction. The CTPS reaction proceeds via the high energy intermediate UTP-4-phosphate (UTP-4-P). Four novel analogues of uridine-4-phosphate (U-4-P) and 3-deazauridine-4-phosphate (3-deazaU 4-P) were synthesized in which the labile phosphate ester oxygen was replaced with a methylene and difluoromethylene group. The methylene analogue of U-4-P, compound 1, was prepared by a reaction of the sodium salt of tert-butyl diethylphosphonoacetate with protected, 4-O-activated uridine followed by acetate deprotection and decarboxylation. It was found that this compound undergoes relatively facile dephosphonylation presumably via a metaphosphate intermediate. The difluoromethylene derivative, compound 2, was prepared by electrophilic fluorination of protected 1. This compound was stable and did not undergo dephosphonylation. Synthesis of the methylene analogue of 3-deazaU-4-P, compound 3, was achieved by ribosylation of protected 4-(phosphonomethyl)-2 hydroxypyridine. Electrophilic fluorination was also employed in the preparation of protected 4-(phosphonodifluoromethyl)-2-hydroxypyridine which was used as the key building block in the synthesis of difluoro derivative 4. These compounds represent the first examples of a nucleoside in which the base has been chemically modified with a methylene or difluormethylenephosphonate group. PMID- 17137370 TI - Total synthesis of the cytostatic marine natural product dibromophakellstatin via three-component imidazolidinone anellation. AB - The tetracyclic pyrrole-imidazole alkaloid dibromophakellstatin from the marine sponge Phakellia mauritiana has been synthesized within seven steps from pyrrole in an 18% overall yield. The key step is a three-component assembly of a tricyclic enamide, a nitrene, and a carbamoyl building block, affording the imidazolidinone ring of dibromophakellstatin in one step. Notably, it is possible to employ the reagent EtO2CNHOTs in a double function as a source of the electrophilic nitrene and of a dipolar carbamoyl component. Use of debrominated precursor dipyrrolopyrazinones leads to much higher anellation yields and allowed us to develop a second generation synthesis. The cytostatic activity of dibromophakellstatin is confirmed. PMID- 17137371 TI - Total synthesis of natural and unnatural lamellarins with saturated and unsaturated D-rings. AB - Twenty-eight natural and unnatural lamellarins with either a saturated or an unsaturated D-ring were synthesized according to our developed synthetic route. The key step involved the Michael addition/ring closure (Mi-RC) of the benzyldihydroisoquinoline and alpha-nitrocinnamate derivatives, which provided the 2-carboethoxypyrrole intermediates in moderate to good yields (up to 78% yield). Subsequent hydrogenolysis/lactonization furnished lamellarins with a saturated D-ring in excellent yields (up to 93% yield). DDQ oxidation of the saturated lamellarin acetates led directly to the corresponding unsaturated analogues in 54-95% yield. In addition, only two steps in our developed strategy require column chromatography. PMID- 17137372 TI - Stereoelectronic effects on 1H nuclear magnetic resonance chemical shifts in methoxybenzenes. AB - Investigation of all O-methyl ethers of 1,2,3-benzenetriol and 4-methyl-1,2,3 benzenetriol (3-16) by 1H NMR spectroscopy and density-functional calculations disclosed practically useful conformational effects on 1H NMR chemical shifts in the aromatic ring. While the conversion of phenol (2) to anisole (1) causes only small positive changes of 1H NMR chemical shifts (Delta delta < 0.08 ppm) that decrease in the order Hortho > Hmeta > Hpara, the experimental O-methylation induced shifts in ortho-disubstituted phenols are largest for Hpara, Delta delta equals; 0.19 +/- 0.02 ppm (n = 11). The differences are due to different conformational behavior of the OH and OCH3 groups; while the ortho-disubstituted OH group remains planar in polyphenols due to hydrogen bonding and conjugative stabilization, the steric congestion in ortho-disubstituted anisoles outweighs the conjugative effects and forces the Ar-OCH3 torsion out of the ring plane, resulting in large stereoelectronic effects on the chemical shift of Hpara. Conformational searches and geometry optimizations for 3-16 at the B3LYP/6-31G** level, followed by B3LYP/6-311++G(2d,2p) calculations for all low-energy conformers, gave excellent correlation between computed and observed 1H NMR chemical shifts, including agreement between computed and observed chemical shift changes caused by O-methylation. The observed regularities can aid structure elucidation of partly O-methylated polyphenols, including many natural products and drugs, and are useful in connection with chemical shift predictions by desktop computer programs. PMID- 17137373 TI - Proline promoted synthesis of ring-fused homodimers: self-condensation of alpha,beta-unsaturated aldehydes. AB - 1,2,4-Trisubstituted cyclohexadienals can be prepared synthetically by self condensation of beta-methyl substituted alpha, beta-unsaturated aldehydes. While molecules with this structural scaffold have been observed in nature, the biological roles of these compounds have yet to be thoroughly investigated. Here we investigate the use of L-proline and its derivatives to effect synthesis of these ring-fused homodimers. The scope of this reaction is investigated with different substrates and proline derivatives. Mechanistic hypotheses are put forth supported by NMR and mass spectrometry studies. The method will enable diversification of this scaffold in sufficient quantities for biological investigations. PMID- 17137374 TI - The synthesis of new 60 degrees organometallic subunits and self-assembly of three-dimensional M3L2 trigonal-bipyramidal cages. AB - By employing the coupling reaction of trans-PtI2(Et3)2 with C-H bonds in alkynes as the key step, two new 60 degrees organometallic subunits with different size from 3,6-dibromophenanthrene were prepared in reasonable yields. The X-ray structures of both di-Pt(II) diiodide complexes showed that they were indeed suitable candidates for 60 degrees building units. By utilizing these novel linkers as ditopic acceptor subunits, three M3L2 trigonal-bipyramidal (TBP) cages were formed. All three TBP cages were characterized with multinuclear NMR and electrospray ionization mass spectrometry (ESI-MS) along with element analysis. PMID- 17137375 TI - N-acyl triazenes as tunable and selective chemodosimeters toward cyanide ion. AB - A novel type of chemodosimeters has been developed on the basis of a displacement reaction. N-Acyl-triazenes are found to be highly selective and tunable chemodosimeters toward cyanide. When N-acetyl-triazene 1a was titrated with various anions (-CN, F-, Cl-, AcO-, H2PO4-, -SCN, ClO4-, and HSO4-) in acetonitrile, significant absorption changes (from colorless to deep purple) resulted in the cases of -CN and F-, and small changes in the cases of AcO- and H2PO4-. N-Isopropanoyl-triazene 1b showed significant response only toward -CN, weaker response toward F-, and little response toward other anions in acetonitrile. Both of the triazenes 1a and 1b responded only to -CN in methanol water, because of the analyte's strong nucleophilicity toward the acyl group and weak hydrogen-bonding ability as compared to other anions examined. The N-acyl triazenes can be used for the naked eye detection of cyanide. PMID- 17137376 TI - Synthesis, optical, and electrochemical properties of a new family of dendritic oligothiophenes. AB - A new class of semi-flexible dendrimers with oligothiophene (OT) arms up to the third generation have been synthesized and investigated. The synthetic methods employed include a combination of palladium-catalyzed Stille cross-coupling reactions for oligothiophenes, Sonogashira cross-coupling reactions for building blocks, and carbodiimide-mediated esterification for building up the various dendrimers. The optical and electrochemical properties of this series of oligothiophenes-based dendrimers are shown to be strongly influenced by their morphologies as demonstrated by their pronounced solvatochromic and thermochromic responses under different environmental conditions. Introducing rigid oligothiophene arms to shape non-persistent ester-linked dendrimers causes higher generation dendrimers (G2 and G3) to exhibit solvatochromism and thermochromism, while their oligomeric counterpart (3b) and lower generation (G1) analogue do not. Spectroscopic changes due to both intramolecular and intermolecular aggregations are observed. PMID- 17137377 TI - Enantioselective enzymatic reductions of sterically bulky aryl alkyl ketones catalyzed by a NADPH-dependent carbonyl reductase. AB - The enantioselective reductions of aryl alkyl ketones, ArC(O)R, with a diverse number of alkyl groups have been achieved with an isolated carbonyl reductase from Sporobolomyces salmonicolor. Of special interest is the observation that ketones with sterically bulky alkyl groups could be reduced to the corresponding alcohols in excellent optical purity. An unusual alkyl chain-induced enantiopreference reversal was observed but was shown to be consistent with the enzyme-substrate docking calculations. PMID- 17137378 TI - Synthesis of benzophenone-containing fatty acids. AB - Syntheses of new benzophenone-containing fatty acids (FABPs) 1, 5, and 6 and a new route to FABP 3 are described. Combined with the known 2 and 4, these FABPs comprise a set of photoactivatable fatty acid analogues with the crosslinking site at defined distances from the carboxylic acid hydroxyl group oxygen atoms ranging from 7.9 to 25.0 A. PMID- 17137379 TI - An amphiphilic molecular basket sensitive to both solvent changes and UV irradiation. AB - A molecular basket was obtained by linking four cholate units to a cone-shaped calix[4]arene scaffold through azobenzene spacers. The molecule turns its polar faces inward in nonpolar solvents to bind polar molecules such as sugar derivatives. In polar solvents, the nonpolar faces turn inward, allowing the binding of hydrophobic guests such as pyrene. The molecule can also respond to UV irradiation by trans-cis isomerization of the azobenzene spacers. Response toward both solvents and UV light is fully reversible. PMID- 17137380 TI - Total synthesis of indole alkaloid (+/-)-subincanadine F via SmI2)-mediated ring opening and bridge-forming Mannich reaction. AB - The first total synthesis of (+/-)-subincanadine F, a bioactive indole alkaloid structurally featuring a 1-azabicyclo[4.3.1]decane unit, has been realized from 1 (para-methoxybenzyl)tryptamine in six steps. The bridge-containing tetracyclic framework of subincanadine F was efficiently assembled by a SmI2-mediated ring opening followed by an acid-mediated Mannich reaction. In addition, the tetracyclic ketoester 6, a key intermediate potentially useful for synthesizing structurally related indole alkaloids as well, was obtained in one step from alpha,beta-diketoester 5. PMID- 17137381 TI - Palladium-catalyzed sonogashira and hiyama reactions using phosphine-free hydrazone ligands. AB - Palladium/copper-catalyzed Sonogashira cross-coupling reaction of aryl halides with a variety of terminal alkynes under amine-free conditions in dimethylformamide (DMF) at 80 degrees C gave internal arylated alkynes using PdCl2(MeCN)2 with phosphine-free hydrazone 2a as a ligand and CuI as the cocatalyst in good yields. We also found PdCl2/hydrazone ligand 1d in PhMe at 80 degrees C was a phosphine-free efficient catalyst system for a Hiyama cross coupling reaction of aryl bromides with aryl(trialkoxy)silanes in good yields. PMID- 17137382 TI - 2-Hydroxymethylboronate as a reagent to detect carbohydrates: application to the analysis of the formose reaction. AB - 2-Hydroxymethylphenylboronate is described as a reagent that converts neutral 1,2 diols, as found in simple carbohydrates, into 1:1 anionic complexes that are easily detected by Fourier transform ion cyclotron resonance mass spectrometry. The value of this reagent was demonstrated through its application to analyze complex mixtures of carbohydrates formed in the formose process, often cited as a way that biologically significant carbohydrates might have been generated from formaldehyde under prebiotic conditions. Coupled with isotope studies, the reagent shows that the simplest autocatalytic cycle for the consumption of formaldehyde in this process cannot account for the bulk consumption of formaldehyde. PMID- 17137383 TI - Ru-catalyzed stereoselective addition of imides to alkynes. AB - A catalyst system formed in situ from bis(2-methylallyl)cycloocta-1,5 dieneruthenium(II) ((cod)Ru[met]2), a phosphine, and scandium(III) trifluoromethanesulfonate (Sc(OTf)3) was found to efficiently catalyze the anti Markovnikov addition of imides to terminal alkynes, allowing mild and atom economic synthesis of enimides. Depending on the phosphine employed, both the (E) and the (Z)-isomer can be accessed stereoselectively. PMID- 17137384 TI - Chiral diols: a new class of additives for direct aldol reaction catalyzed by L proline. AB - Nine C2 symmetric diols have been examined as additives in the L-proline catalyzed direct aldol reaction with significant improvement in enantioselectivity, conversion efficiency, and yield. Loading of 1 mol % of (S) BINOL leads to the desired products in up to 98% ee and 90% yield. A transition state is proposed. PMID- 17137385 TI - Synthesis of loliolide, actinidiolide, dihydroactinidiolide, and aeginetolide via cerium enolate chemistry. AB - Loliolide, aeginetolide, actinidiolide, and dihydroactinidiolide were synthesized in racemic form from a single common intermediate, prepared through the 1,2 addition of the cerium enolate of ethyl acetate to 2,6,6-trimethylcylohexenone. PMID- 17137386 TI - An improved method for the protection of carboxylic acids as 1,1-dimethylallyl esters. AB - 1,1-Dimethylallyl (DMA) esters of various N-protected amino acids have been synthesized using prenyldimethylsulfonium tetrafluroborate, a reagent that can be readily made and stored, in conjunction with catalytic CuBr. These reactions were complete within several hours and afforded DMA esters in high yields. As has been previously shown in our group, DMA esters represent a palladium-labile proctecting group for carboxylic acids that resists nucleophilic attack as a tert butyl ester would. PMID- 17137387 TI - Enantioselective synthesis of gamma-aryl-gamma-butyrolactones by sequential asymmetric epoxidation, ring expansion, and Baeyer-Villiger oxidation. AB - This paper describes an enantioselective synthesis of gamma-butyrolactones, using the N-tolyl-substituted oxazolidinone-containing ketone as catalyst and Oxone as oxidant via a sequential asymmetric epoxidation of benzylidenecyclopropanes, ring expansion, and Baeyer-Villiger oxidation. Up to 91% ee was obtained. Optically active cyclobutanones can also be obtained by suppressing the Baeyer-Villiger oxidation with use of more ketone catalyst and less Oxone. PMID- 17137388 TI - An efficient base-free N-arylation of imidazoles and amines with arylboronic acids using copper-exchanged fluorapatite. AB - N-Arylation of imidazoles and amines with arylboronic acids was accomplished with copper-exchanged fluorapatite (CuFAP) in methanol at room temperature. The products N-arylimidazoles and N-arylamines were isolated in good to excellent yields. A variety of arylboronic acids were converted to the corresponding N arylimidazoles and N-arylamines, demonstrating the versatility of the reaction. PMID- 17137389 TI - Chemoselective protection of solid-phase compatible Fmoc-phosphinic building blocks. AB - An efficient four-step synthetic strategy able to fully discriminate hydroxyphosphinyl and carboxylic groups of Fmoc-phosphinic building blocks and related analogues has been developed. The proposed method applies selective acidic removal of the phenacyl (Pac) group from the hydroxyphosphinyl functionality and protection by the 1-adamantyl (Ad) group. Reductive removal of the Pac group from the carboxylic functionality leads to Fmoc-protected phosphinic pseudodipeptidic units suitable for phosphinic peptide and library development using solid-phase peptide synthesis (SPPS). PMID- 17137390 TI - Free-radical approaches to stemoamide and analogues. AB - Two approaches allowing access to the tricyclic stemona backbone are presented. Both approaches rely on a free-radical cyclization reaction as the key step. In the formal synthesis of (+/-)-stemoamide, the construction of the A ring of the natural product was achieved via a 5-exo-trig radical cyclization with atom transfer. The two diastereoisomers issuing from this cyclization showed different reactivity while forming the seven-membered ring of the final product. In the second part of this study, a 7-exo-trig free radical cyclization was realized allowing access to the (+/-)-9,10-bis-epi-stemoamide. This reaction was highly stereoselective and allowed the control of three of the four contiguous stereocenters present in the molecule. PMID- 17137391 TI - An unusual beta-vinyl effect leading to high efficiency and enantioselectivity of the amidase, nitrile biotransformations for the preparation of enantiopure 3 arylpent-4-enoic acids and amides and their applications in synthesis. AB - Biotransformations of 3-arylpent-4-enenitriles catalyzed by Rhodococcus erythropolis AJ270, a nitrile hydratase/amidase-containing microbial whole-cell catalyst were studied, and an unusual beta-vinyl effect of the substrate on the biocatalytic efficiency and enantioselectivity of the amidase was observed. While 3-arylpent-4-enenitriles and 3-phenylpentanenitrile were efficiently hydrated by the action of the less R-enantioselective nitrile hydratase, the amidase showed greater activity and higher enantioselectivity against 3-arylpent-4-enoic acid amides than 3-arylpentanoic acid amides. Under very mild conditions, nitrile biotransformations provided an efficient synthesis of highly enantiopure (R)-3 arylpent-4-enoic acids and (S)-3-arylpent-4-enoic acid amides, and their applications were demonstrated by the synthesis of chiral gamma-amino acid, 2 pyrrolidinone, and 2-azepinone derivatives. PMID- 17137392 TI - Amberlite IRA900N3 as a new catalyst for the azidation of alpha,beta-unsaturated ketones under solvent-free conditions. AB - Amberlite IRA900N3 is an excellent organocatalyst for the azidation of alpha,beta unsaturated ketones with trimethylsilyl azide under solvent-free conditions. By avoiding the use of metallic species and of the organic reaction medium, the procedure is a green tool for the preparation of beta-azido ketones under mild conditions with yields from good to excellent. The catalyst can be recovered and re-used with no loss of its efficiency. PMID- 17137393 TI - One-Step conversion of pyridine N-oxides to tetrazolo[1,5-a]pyridines. AB - Pyridine N-oxides were converted to tetrazolo[1,5-a]pyridines in good to excellent yield by heating in the presence sulfonyl or phosphoryl azides and pyridine in the absence of solvent. Various sulfonyl and phosphoryl azides were screened for reactivity under a standard set of conditions. Diphenyl phosphorazidate was the most convenient reagent and gave high yields. Reaction optimization, scope, and scalability are discussed. PMID- 17137394 TI - Complexity-enhancing acid-promoted rearrangement of tricyclic products of tandem Ugi 4CC/intramolecular Diels-Alder reaction. AB - The unexpected rearrangement of 2,3,7,7a-tetrahydro-3a,6-epoxyisoindol-1-one products of the tandem Ugi 4CC/intramolecular Diels-Alder reaction in 85% H3PO4 was discovered to provide diastereomerically pure tricyclic bislactam lactone containing natural product-like products in high yield. Mechanistic rationale for the observed rearrangement was proposed and has been tentatively confirmed by additional experiments. PMID- 17137395 TI - Superior reactivity of thiosemicarbazides in the synthesis of 2-amino-1,3,4 oxadiazoles. AB - A facile and general protocol for the preparation of 2-amino-1,3,4-oxadiazoles is reported. This method relies on a tosyl chloride/pyridine-mediated cyclization of a thiosemicarbazide, which is readily prepared by acylation of a given hydrazide with the appropriate isothiocyanate. Cyclization of the thiosemicarbazide consistently outperforms the analogous semicarbazide cyclization under these conditions, for 18 distinct examples. Utilizing this protocol, we have prepared 5 alkyl- and 5-aryl-2-amino-1,3,4-oxadiazoles in 78-99% yield. PMID- 17137396 TI - Carbolithiation of diphenylacetylene as a stereoselective route to (Z)-tamoxifen and related tetrasubstituted olefins. AB - Carbolithiation of diphenylacetylene can be exploited to generate (E)-1-lithio 1,2-diphenylalkyl-1-enes which can be reacted in situ with triisopropylborate to stereoselectively provide (E)-1,2-diphenyl-1-alkylene boronic acids. These tetrasubstituted vinylboronic acids served as versatile intermediates for the generation of tetrasubstituted olefins with retention of stereochemistry. The application of this method for the stereoselective synthesis of (Z)-tamoxifen and related analogues is described. PMID- 17137397 TI - A cleavable amino-thiol linker for reversible linking of amines to DNA. AB - A cleavable heterobifunctional cross-linker for the reversible conjugation of amines to thiol-modified DNA has been developed and tested. The succinimidyl 2 (vinylsulfonyl)ethyl carbonate (SVEC) was prepared in three steps and tested for its ability to react with amines and thiols. The linker was efficient for binding leucine to a thiol-modified DNA sequence and for releasing the amino acid at pH 11.8. PMID- 17137400 TI - Potential of aromatase inhibitors for ovulation and superovulation induction in infertile women. AB - For almost half a century, the first-line treatment for ovulation induction in cases of anovulation, unexplained infertility, or mild male factor has been clomifene (clomiphene citrate). Clomifene is an effective and safely used oral agent, but is known to have relatively common antiestrogenic endometrial and cervical mucous adverse effects that could prevent pregnancy in the face of successful ovulation. In addition, there is a significant risk of multiple pregnancies with clomifene compared with natural cycles. These drawbacks are mainly a result of the extended antiestrogenic effect of clomifene as a result of its accumulation in the body (clomifene isomers have a half-life of several days up to few weeks). Because of these problems, we proposed the concept of aromatase inhibition as a new method of ovulation induction that could avoid many of the adverse effects of clomifene. Over the last few years several published studies, both controlled and noncontrolled, compared clomifene and treatment with aromatase inhibitors (AIs), either alone or in combination with gonadotropins, for ovulation induction or augmentation. These studies found AIs as effective as clomifene in inducing ovulation, with the major advantage of absence of any antiestrogenic adverse effects. Several other major advantages of AIs include the lower serum estrogen production per developing follicle resulting in more physiological estrogen levels around the time of ovulation and good pregnancy rates with a lower incidence of multiple pregnancy than with clomifene. When combined with gonadotropins for assisted reproductive technologies, AIs reduce the dose of gonadotropins required for optimal follicle recruitment and improve the response to gonadotropin stimulation in poor responders. Such preliminary evidence suggests that AIs may replace clomifene in the future because of similar efficacy with a reduced adverse-effect profile. However, we believe that definitive studies in the form of randomised controlled trials comparing clomifene with AIs are needed. PMID- 17137401 TI - Targeted anti-cancer therapies for renal cancer. AB - In the past several years, significant advances in the underlying biological mechanisms of renal cell cancer, particularly the role of tumour angiogenesis, have permitted the design of molecularly targeted therapeutics. For this review, single-agent therapies inhibiting the following different targets were identified in the published literature: epithelial growth factor receptor, vascular endothelial growth factor receptor, basic fibroblast growth factor receptor, platelet-derived growth factor, nuclear factor-kappabeta, the mammalian target of rapamycin (mTOR) pathway, raf kinase pathway and tyrosine kinase pathway. Distinct fields of clinical research have emerged--monoclonal antibodies, small molecules, nanopeptides and immunomodulators. All therapies demonstrated acceptable toxicity profiles. Clinical benefit was assessed on the basis of the reported criteria for each study, and antitumour response (regression or delay in progression-free survival) ranged from 5% to 71%. On the basis of the limited studies to date, targeted therapies offer the greatest clinical benefit in the management of this malignancy, although additional basic research is still warranted to further improve clinical outcome. PMID- 17137402 TI - Polycythaemia vera and essential thrombocythaemia: current treatment strategies. AB - Polycythaemia vera (PV) and essential thrombocythaemia (ET) are classified as Philadelphia-negative chronic myeloproliferative diseases. Both PV and ET are rare diseases, but the prevalence is high. Patients who have not been treated for the diseases are at great risk of morbidity and mortality as a result of thrombohaemorrhagic events. However, if patients have been well treated, their prognosis is good and life-expectancy approaches normal. This article provides diagnostic tools and flowcharts for treatment of PV and ET. Treatment of PV and ET should be risk-adjusted and individualised. Low-dose aspirin is recommended as an antiaggregative drug in both diseases. For PV, phlebotomy to control a haematocrit at <0.45 is the cornerstone in treatment, and treatment with hydroxycarbamide (hydroxyurea) or interferon (IFN)-alpha is added to reduce hypermetabolic symptoms or splenomegaly becoming cytoreductive. In ET, hydroxycarbamide and anagrelide are the most used drugs, and anagrelide may also be added in PV to reduce thrombocytosis. IFNalpha is the only myelosuppressive treatment available during pregnancy. Current controversies regarding treatment illustrate the need for more randomised clinical trials. Demonstration of over expression of the PV-1 gene and in particular the JAK-2 mutation will be novel diagnostic criteria and may have an impact for future therapy of both PV and ET. PMID- 17137403 TI - Current pharmacological options for the management of primary hyperparathyroidism. AB - Drugs for treating primary hyperparathyroidism can be divided into two main groups: (i) antiresorptive drugs that inhibit the increased bone turnover, which can be divided into estrogen-like compounds (estrogen, oral contraceptives and selective estrogen receptor modulators [SERMs]), bisphosphonates and calcitonin; and (ii) drugs that interfere with parathyroid hormone (PTH) secretion (currently only cinacalcet is available). No drugs that interfere with PTH action are currently available. Available studies suggest that all classes of drugs are able to lower serum calcium levels. However, calcitonin does so only temporarily. Estrogen-containing compounds (hormone replacement therapy) may be less attractive because of the potential risk of breast cancer, cardiovascular disease and deep vein thromboembolism. Oral contraceptives have not been shown to be able to prevent fractures in the general population, and no data are available on their effect in women with primary hyperparathyroidism. The only SERM marketed for hyperparathyroidism is raloxifene and this has not been associated with an increased risk of breast cancer and cardiovascular diseases, and has been shown to be able to prevent vertebral fractures in postmenopausal women with osteoporosis. Two small trials suggest that raloxifene may increase bone mineral density (BMD) and decrease serum calcium levels in patients with primary hyperparathyroidism. Bisphosphonates have been shown to decrease serum calcium and increase BMD in patients with primary hyperparathyroidism, but PTH levels may increase. Cinacalcet effectively induces a sustained decrease in serum calcium and PTH for up to 1 year. However, BMD does not seem to increase. No data on hard endpoints such as fractures, kidney stones, cardiovascular disease etc. are available for any of the drugs available for the treatment of primary hyperparathyroidism. PMID- 17137405 TI - Efficacy and safety of budesonide and formoterol in one pressurised metered-dose inhaler in adults and adolescents with moderate to severe asthma: a randomised clinical trial. AB - BACKGROUND: Inhaled corticosteroids (ICSs) are the preferred maintenance therapy for adults and children with mild, moderate and severe persistent asthma, with the addition of a long-acting beta(2)-adrenoceptor agonist to ICS therapy recommended for patients with moderate or severe persistent asthma. The efficacy and safety of the combination of budesonide and formoterol delivered via dry powder inhaler (DPI) is well documented. OBJECTIVE: To compare the efficacy and safety of budesonide/formoterol pressurised metered-dose inhaler (budesonide/formoterol pMDI; Symbicort pMDI, AstraZeneca LP, Wilmington, DE, USA) with budesonide pMDI (Pulmicort pMDI, Astra [corrected] Zeneca, Lund, Sweden), formoterol DPI (Oxis Turbuhaler, AstraZeneca, Lund, Sweden), budesonide plus formoterol in separate inhalers (budesonide pMDI + formoterol DPI) and placebo. STUDY DESIGN: This was a 12-week randomised, double-blind, double-dummy, placebo controlled study. SETTING: This multicentre study was conducted in the respiratory specialty clinical practice setting. PATIENTS: The study included 596 patients > or =12 years of age with moderate to severe persistent asthma previously receiving ICSs. INTERVENTIONS: After 2 weeks on budesonide pMDI 80 microg x two inhalations (160 microg) twice daily, patients received budesonide/formoterol pMDI 160 microg/4.5 microg x two inhalations (320 microg/9 microg); budesonide pMDI 160 microg x two inhalations (320 microg) + formoterol DPI 4.5 microg x two inhalations (9 microg); budesonide pMDI 160 microg x two inhalations (320 microg); formoterol DPI 4.5 microg x two inhalations (9 microg); or placebo twice daily. MAIN OUTCOME MEASURES: There were two prespecified primary efficacy variables: mean change from baseline in morning predose forced expiratory volume in 1 second (FEV(1)), obtained approximately 12 hours after the most recent administration of study medication at home and immediately before the next administration of study medication at the clinic; and mean change from baseline in 12-hour FEV(1), assessed as the average change in FEV(1) from serial spirometry over the 12-hour period after administration of the morning dose of study medication at the clinic. RESULTS: Mean changes from baseline in morning predose FEV(1) at end of treatment were greater (p < or = 0.049) with budesonide/formoterol pMDI (0.19L) versus budesonide pMDI (0.10L), formoterol DPI (-0.12L) and placebo (-0.17L). Mean changes from baseline in 12-hour FEV(1) were greater (p < or = 0.001) with budesonide/formoterol pMDI after 1 day (0.37L), 2 weeks (0.34L) and at end of treatment (0.37L) versus budesonide pMDI (0.11, 0.15 and 0.15L) and placebo (0.09, -0.03 and -0.03L), and after 2 weeks and at end of treatment versus formoterol DPI (0.19 and 0.17L). Fewer (p < or = 0.025) patients receiving budesonide/formoterol pMDI versus monoproducts or placebo met worsening asthma criteria. Results were similar in the budesonide/formoterol pMDI group and the budesonide pMDI + formoterol DPI group on all measures. All treatments were well tolerated with similar safety profiles. CONCLUSIONS: In this population, twice-daily budesonide/formoterol pMDI provides asthma control significantly greater than the monocomponents or placebo and comparable with budesonide pMDI + formoterol DPI. Safety profiles were similar for all treatments. PMID- 17137404 TI - Blood pressure lowering for the prevention and treatment of diabetic kidney disease. AB - The current pandemic of diabetes mellitus will inevitably be followed by an epidemic of chronic kidney disease. It is anticipated that 25-40% of patients with type 1 diabetes and 5-40% of patients with type 2 diabetes will ultimately develop diabetic kidney disease. The control of blood pressure represents a key component for the prevention and management of diabetic nephropathy. There is a strong epidemiological connection between hypertension in diabetes and adverse outcomes in diabetes. Hypertension is closely linked to insulin resistance as part of the 'metabolic syndrome'. Diabetic nephropathy may lead to hypertension through direct actions on renal sodium handling, vascular compliance and vasomotor function. Recent clinical trials also support the utility of blood pressure reduction in the prevention of diabetic kidney disease. In patients with normoalbuminuria, transition to microalbuminuria can be prevented by blood pressure reduction. This action appears to be significant regardless of whether patients have elevated blood pressure or not. The efficacy of ACE inhibition appears to be greater than that achieved by other agents with a similar degree of blood pressure reduction; although large observational studies suggest the risk of microalbuminuria may be reduced by blood pressure reduction, regardless of modality. In patients with established microalbuminuria, ACE inhibitors and angiotensin receptor antagonists (angiotensin receptor blockers [ARBs]) consistently reduce the risk of progression from microalbuminuria to macroalbuminuria, over and above their antihypertensive actions. The clinical utility of combining these strategies remains to be established. In patients with overt nephropathy, blood pressure reduction is associated with reduced urinary albumin excretion and, subsequently, a reduced risk of renal impairment or end stage renal disease. In addition to actions on systemic blood pressure, it is now clear that ACE inhibitors and ARBs also reduce proteinuria in patients with diabetes. This anti-proteinuric activity is distinct from other antihypertensive agents and diuretics. Although there is a clear physiological rationale for blockade of the renin angiotensin system, which is strongly supported by clinical studies, to achieve the optimal lowering of blood pressure, particularly in the setting of established diabetic renal disease, a number of different antihypertensive agents will always be needed. In the end, the choice of agents should be individualised to achieve the maximal tolerated reduction in blood pressure and albuminuria. Ultimately, no matter how it is achieved, so long as it is achieved, renal risk can be reduced by agents that lower blood pressure and albuminuria. PMID- 17137406 TI - Sunitinib. AB - Sunitinib and its active metabolite (SU012662) are selective inhibitors of multiple receptor tyrosine kinases associated with tumour growth and angiogenesis. The clinical efficacy of oral sunitinib has been demonstrated in patients with advanced gastrointestinal stromal tumours (GIST). In a phase III, randomised, double-blind, placebo-controlled, multicentre trial in patients with metastatic and/or unresectable GIST following unsuccessful imatinib therapy, the median time to tumour progression and median progression-free survival time were > or =4-fold longer in patients receiving sunitinib 50 mg/day than in those receiving placebo, in 6-week cycles consisting of 4 weeks of treatment followed by a 2-week rest period. Sunitinib also exhibited antitumour activity in patients with advanced renal cell carcinoma (RCC) following unsuccessful cytokine therapy. In two multicentre, single-arm, phase II clinical trials in patients with cytokine-refractory metastatic RCC, partial responses were reported in 40% and 43% of patients receiving sunitinib 50 mg/day for 4 weeks followed by 2 weeks without treatment in 6-week cycles; 27% and 22% of patients achieved stable disease for > or =3 months. Sunitinib was more effective than interferon-alpha as a first-line therapy in patients with metastatic RCC. In a large, well designed, phase III trial in previously untreated patients, progression-free survival was significantly longer in patients receiving sunitinib 50 mg/day in 6-week cycles (4 weeks of treatment followed by a 2-week rest period) compared with those receiving interferon-alpha 9MU three times weekly (47.3 vs 24.9 weeks). In general, sunitinib was well tolerated in patients with GIST and RCC, with adverse events usually being of mild or moderate severity. PMID- 17137409 TI - Tacrolimus: in heart transplant recipients. AB - Tacrolimus is a calcineurin inhibitor recently approved in the US and throughout the EU for the prevention of allograft rejection in heart transplant recipients. It is commonly administered orally for long-term immunosuppression. The incidence of mild to severe acute rejection in the first 6 months following heart transplantation was significantly lower in tacrolimus recipients than in ciclosporin recipients (54% vs 66%) in a large, phase III trial conducted in Europe. A large, phase III trial conducted in the US did not show a significant difference between tacrolimus and ciclosporin in the incidence of severe rejection or haemodynamic compromise rejection requiring treatment within the first 6 months post-transplant (22% vs 32%), but did show a significant difference in the incidence at 1 year (23% vs 37%). In phase III trials, 1-year patient survival was similar between tacrolimus and ciclosporin recipients in the EU (93% vs 92%) and the US (95% vs 90%). Tacrolimus was shown to be effective in the prevention of rejection in paediatric and African American heart transplant recipients. The tolerability profile of tacrolimus in heart transplant recipients was broadly similar to that of ciclosporin, although tacrolimus was usually associated with lower incidences of post-transplant hypertension and dyslipidaemia. PMID- 17137413 TI - A uniqueness to personal threat (UPT) hypothesis: how similarity affects perceptions of susceptibility and severity in risk assessment. AB - In the literature, perceived risk is conceptualized as perceptions of both susceptibility and severity. Optimistic bias, the belief that one is less vulnerable than others to various diseases, is based primarily on assessments of personal susceptibility. In this article, the authors ask how perceptions of relative severity/susceptibility differ as a function of perceived similarity between oneself and a referent. Based on predictions from the optimistic bias literature, Study 1 (N = 67) manipulated similarity and measured both susceptibility and severity. As anticipated, ratings of susceptibility were negatively correlated with similarity. Ratings of severity, however, were unexpected in that they were highest for a similar referent. The uniqueness of personal threat hypothesis was proposed as an explanation for this finding, and this hypothesis was tested in Study 2 (N = 121). Results were mostly supportive. Findings indicated that individuals strategically use susceptibility and severity ratings to modulate their assessment of overall risk and personal identity. PMID- 17137414 TI - Theoretical approaches to entertainment education campaigns: a subaltern critique. AB - Entertainment education (E-E) is one of the most widely discussed areas in current scholarship on international health communication. In fact, much of the health communication scholarship has been historically dominated by E-E efforts directed at subaltern spaces. This article applies a subaltern studies perspective to interrogate the location of agency of the subaltern participant in the dominant E-E discourse. Based on a critical approach to E-E, the article offers points of departure for studying health communication in subaltern spaces. Subaltern voices point toward alternative definitions of problems beyond the narrow realm of problems defined by the core actors in E-E. Finally, alternative positions are suggested for applying participatory communication in engaging with subaltern participants for problem definition and solution development. PMID- 17137415 TI - Review of interactive safer sex Web sites: practice and potential. AB - The Internet is increasingly being viewed as a health promotion tool with enormous potential. However, this potential cannot be realized if Web sites do not utilize the features that make the Internet a "hybrid" mass and interpersonal communication medium. The purpose of this study was to examine interactive safer sex Web sites on a number of dimensions. A comprehensive search that included Internet search engines, links from well-known sites, and previously published reviews yielded 21 Web sites that met criteria. Web sites were coded on dimensions including targeting of the Web sites, safer sex messages presented, theoretical strategies utilized, interactivity, and other characteristics. Results indicate that a moderate amount of targeting of Web sites exists, especially on age group (e.g., teenagers); the most prevalent safer sex messages were to "use condoms" and "be sexually abstinent"; raising the perceived threat of sexually transmitted diseases and HIV was the most prevalent theoretical strategy used to motivate safer sex; and finally, a moderate amount of interactivity was found on the Web sites, with most Web sites containing 4 or 5 features out of 15 features examined. Evidence that Web sites were tailoring information or messages to individuals was not found. Implications of these results for improving safer sex Web sites and developing interventions online are discussed. PMID- 17137416 TI - StarvingforPerfect.com: a theoretically based content analysis of pro-eating disorder Web sites. AB - Like traditional media, information on the World Wide Web may encourage both healthy and unhealthy behaviors. This study reports on the content analysis of a particular genre of Web site that promotes unhealthy behaviors: pro-eating disorder Web sites. Framed in message design theory, the results of this study indicate that messages on pro-eating disorder Web sites promote response efficacy in continuing disordered behaviors, but messages promoting severity and susceptibility to weight gain and self-efficacy were not common. Given the importance of combining response and self-efficacy messages for maximal effectiveness of messages, the pro-eating disorder sites may have limited effectiveness in effecting behavioral change among site visitors. PMID- 17137417 TI - Age differences in how consumers behave following exposure to DTC advertising. AB - This study was conducted to provide additional evidence on how consumers behave following direct-to-consumer (DTC) advertising exposure and to determine if there are differences in ad-prompted acts (drug inquiry and drug requests) between different age groups (i.e., older, mature, and younger adults). The results suggest that younger, mature, and older consumers are all moved to act by DTC drug ads, but that each age group behaves in different ways. Somewhat surprisingly, age was not predictive of ad-prompted behavior. DTC advertising was no more effective at moving older consumers to behave than their younger counterparts. These results suggest that age does not matter that much when it comes to the "moving power" of prescription drug advertising, even though research indicates that older consumers are more vulnerable to the persuasive effects of communication. PMID- 17137418 TI - The drug resistance strategies intervention: program effects on substance use. AB - This study evaluates the Drug Resistance Strategies (DRS) project, a culturally grounded, communication-based substance use prevention program implemented in 35 middle schools in Phoenix, Arizona. The intervention consisted of 10 lessons taught by the classroom teacher that imparted the knowledge, motivation, and skills needed to resist drug offers. The evaluation used growth modeling to analyze significant differences in average postintervention substance use (alcohol, cigarettes, and marijuana) and growth of use over the course of the study. The study involved 6,298 seventh graders (65% Mexican/Mexican American) who responded to at least 1 of 4 questionnaires (1 pretest and 3 follow-up measures). When compared to a control group, the DRS intervention appeared to significantly limit the increase in the number of students reporting recent substance use, especially alcohol and marijuana use. The multicultural version of the curriculum proved most broadly effective, followed by the version targeting Mexican American youth. The development of a culturally grounded prevention curriculum for Mexican American youth expands the population being served by interventions. Moreover, the success of the multicultural curriculum version, which has the broadest application, provides particular promise, and the article demonstrates how a growth modeling approach can be used to evaluate a communication-based intervention by analyzing changes over time rather than differences between the pretest and posttest scores. PMID- 17137419 TI - Stepping back from social norms campaigns: comparing normative influences to other predictors of health behaviors. AB - Recent health campaigns on college campuses have used a social norms approach, which suggests that one's perceptions of others' attitudes and behaviors are the key components in attitude and behavior change. However, the efficacy of social norms campaigns has been mixed. This study was conducted to assess the relationships among sociodemographics, normative perceptions, and individual attitudes on 3 health behaviors. Students at 2 universities (N = 393) completed questionnaires assessing how these variables related to their consumption of alcohol, tobacco use, and exercise behaviors. Regressions indicated that each of these variables was associated with behavior, but varied independent variables emerged as the salient predictors among behaviors. In several conditions the effect of normative perceptions on behaviors was not significant, a finding in direct opposition to social norms marketing. In all 3 behavioral conditions, the variable accounting for the greatest variance was whether or not the individual liked participating in that particular behavior. Thus, although some social norms marketing may be meeting with success, it may be the case that predicted attitudinal and behavioral changes will not be found when applied across diverse health topics. PMID- 17137420 TI - Healthy research perspectives: incorporating college student experiences with alcohol. AB - College student alcohol consumption is pervasive and problematic at most U.S. college and university campuses. This study focuses on understanding college students who consume high levels of alcohol, providing healthy insight into what have been perceived by researchers as unhealthy behaviors. Researchers conducted 6 mediated focus group discussions. Five strategies are warranted: normalize alcohol consumption, consider alcohol socialization practices used in other cultures, educate parents about the importance of their role, modify current high school alcohol education strategies, and provide student responsibility through peer interaction. PMID- 17137421 TI - Effective message design targeting college students for the prevention of binge drinking: basing design on rebellious risk-taking tendency. AB - This study examined the responses of college students who were exposed to different types of episodic stories related to drinking (gender-consistent vs. gender-inconsistent condition) and their intention to modify risky behavior (binge drinking) based on their rebellious risk-taking tendency. Self-report measures such as intention to modify drinking behavior and reaction to the message were measured. Eighty-two college students between the ages of 19 and 23 years participated in a posttest-only group design experiment. Results suggested that rebellious participants were less afraid of the dangers of binge drinking than those who were low in rebelliousness for the gender-consistent condition. Regardless of the level of rebelliousness, the participants who were in the gender-consistent (increased relevance) condition produced higher recognition scores than those who were in the gender-inconsistent condition. However, the rebellious participants who were in the gender-inconsistent condition exhibited a higher level of intention to change their drinking behavior than did those in the gender-consistent condition. PMID- 17137422 TI - Bibliography. PMID- 17137423 TI - Nuclear receptor structure: implications for function. AB - Small lipophilic molecules such as steroidal hormones, retinoids, and free fatty acids control many of the reproductive, developmental, and metabolic processes in eukaryotes. The mediators of these effects are nuclear receptor proteins, ligand activated transcription factors capable of regulating the expression of complex gene networks. This review addresses the structure and structural properties of nuclear receptors, focusing on the well-studied ligand-binding and DNA-binding domains as well as our still-emerging understanding of the largely unstructured N terminal regions. To emphasize the allosteric interdependence among these subunits, a more detailed inspection of the structural properties of the human progesterone receptor is presented. Finally, this work is placed in the context of developing a quantitative and mechanistic understanding of nuclear receptor function. PMID- 17137424 TI - Men's health and wellbeing: taking up the challenge in Australia. PMID- 17137425 TI - The men in Australia Telephone Survey (MATeS) - lessons for all. PMID- 17137426 TI - Male reproductive health and the environment. PMID- 17137428 TI - Locally acquired infection with Entamoeba histolytica in men who have sex with men in Australia. PMID- 17137429 TI - Men in Australia Telephone Survey (MATeS): predictors of men's help-seeking behaviour for reproductive health disorders. AB - OBJECTIVE: To identify sociodemographic factors associated with help-seeking behaviour for reproductive health disorders in middle-aged and older Australian men. DESIGN: A cross-sectional, population-based, computer-assisted telephone interview exploring sociodemographic factors and general and reproductive health. PARTICIPANTS AND SETTING: Analysis of data from the Men in Australia Telephone Survey (MATeS) of 5990 Australian men aged 40 years and older interviewed between September and December 2003. MAIN OUTCOME MEASURES: Self-reported diagnosis of prostate disease and erectile dysfunction (ED), help-seeking behaviour (including visiting a doctor, prostate-specific antigen testing, treatment of prostate disease, speaking to a health professional about ED and treatment of ED). RESULTS: Age was a significant predictor of all help-seeking behaviour studied, other than treatment for ED. Controlling for all predictor variables, never married status predicted a lower likelihood of visiting a doctor (odds ratio [OR], 0.68 [95% CI, 0.48-0.97]) or speaking to a health professional about ED (OR, 0.44 [95% CI, 0.21-0.93]), while divorced/separated status predicted lower likelihood of having a prostate-specific antigen test (OR, 0.63 [95% CI, 0.50 0.79]). Living in a regional or remote area or being from a non-English-speaking background predicted lower likelihood of receiving treatment for ED (ORs, 0.62 [95% CI, 0.42-0.92] and 0.41 [95% CI, 0.24-0.72], respectively), but did not influence screening for prostate disease. CONCLUSION: Seeking advice or treatment for male reproductive health disorders is predicted by sociodemographic factors specific to different reproductive health problems. As middle-aged and older men do attend doctors, opportunities exist for health professionals to optimise their consultations by routinely discussing reproductive health with all men, to identify under-reported male reproductive health disorders. PMID- 17137431 TI - The association between obesity and the diagnosis of androgen deficiency in symptomatic ageing men. AB - OBJECTIVE: To determine the influence of obesity on the diagnosis of age-related androgen deficiency (AD) in symptomatic men according to current Australian guidelines. DESIGN, SETTING AND PARTICIPANTS: A community-based cohort of healthy ageing men with symptoms suggestive of AD was studied between May 2001 and February 2003. Men were classified as obese or non-obese according to body mass index (BMI) or waist circumference (WC). MAIN OUTCOME MEASURE: Diagnosis of AD according to Endocrine Society of Australia (ESA) guidelines. RESULTS: 223 men aged 54-86 years with mean BMI 27.3 +/- 0.2 kg/m2 (range 20.5-36.2 kg/m2) were recruited; 99 men were obese (BMI > or = 30.0 kg/m2 or WC > or = 102 cm) and 124 men were non-obese. Obese men had lower total testosterone (TT) (12.7 +/- 0.4 v 15.0 +/- 0.4 nmol/L); P < 0.001) and calculated free testosterone (275.7 +/- 7.8 v 299.3 +/- 7.4 pmol/L); P = 0.03) levels than non-obese men. TT levels < 8 nmol/L were recorded in 12% of obese men and 1% of non-obese men. Applying the ESA guidelines for the diagnosis of age-related AD, 15 obese men (15%) and 4 non obese men (3%) were classified as being eligible for androgen therapy supported by the Pharmaceutical Benefits Scheme (PBS); the relative risk in obese men was 1.92 (95% CI, 1.44-2.55; P < 0.001). CONCLUSION: Obesity is an important determinant of serum TT levels in ageing men. Almost one in seven obese men but only one in 30 non-obese men in our study were eligible for PBS-supported androgen therapy according to Australian guidelines. Although obese men are more likely to have biochemical hypoandrogenism, the clinical implications of this remain uncertain. Studies of testosterone therapy in this group of ageing men are needed to determine whether androgen replacement is beneficial. PMID- 17137432 TI - The psychosocial impact of prostate cancer on patients and their partners. AB - OBJECTIVE: To assess the psychosocial impact of the diagnosis of either localised or metastatic prostate cancer (PCA) on patients and their female partners. DESIGN: Observational, prospective study at Time 1 and 6 months later at Time 2 of two groups of couples facing PCA. Time 1 was when patients were first diagnosed with histologically confirmed localised (potentially curable) PCA or metastatic (incurable) PCA. MAIN OUTCOME MEASURES: Depression and anxiety disorders according to the Diagnostic and statistical manual of mental disorders 4th edition (DSM-IV); psychological distress; marital satisfaction. RESULTS: At Time 1, partners had rates of DSM-IV major depression and generalised anxiety disorder twice those of women in the Australian community, and considerably higher than the patients' rates. At Time 2, psychological distress in partners had lessened but that in patients had increased. On the other hand, at Time 2, partners' marital satisfaction had deteriorated. CONCLUSIONS: To be fully effective, interventions aimed at reducing the psychosocial morbidity of PCA must involve both patient and partner, rather than the patient alone. PMID- 17137433 TI - Y chromosome microdeletions: implications for assisted conception. PMID- 17137434 TI - Testosterone: use, misuse and abuse. AB - Testosterone is among the oldest drugs in medicine. It has a long efficacy and safety record for its prime role of androgen replacement therapy in men with androgen deficiency. Testosterone and synthetic analogue androgens have also been used in pharmacological androgen therapy (PAT) to produce androgenic effects on marrow, muscle or bone. Although PAT is increasingly being superseded by newer, more expensive drugs, androgens remain cost-effective in many older applications. Androgen misuse is the systematic over-prescribing for unproven medical indications. Misuse is increasingly evident for male ageing ("andropause") and some other clinical conditions. Further trials for new indications for androgens require reliable safety data, but rising costs may make it increasingly attractive to circumvent the need for evidence by promoting off-label mass marketing. Androgen abuse is the illicit self-administration of often massive doses of androgens for non-medical purposes - notably power sports and body building. In parallel with effective detection reducing androgen abuse in elite sports, more focus is needed on non-sporting cosmetic, recreational and occupational androgen abuse. Despite ongoing androgen misuse and abuse, testosterone remains under-prescribed for younger men with classical androgen deficiency that frequently remains undiagnosed. PMID- 17137435 TI - Men's health: what's a GP to do? AB - Men are at highest risk of cardiovascular disease, chronic lung disease, some cancers, suicide and transport-related injury. An anticipatory approach to men's health in general practice should assess risk for these conditions and offer effective interventions, either to prevent them or manage them early. This requires attention to the barriers, not only to men accessing general practice, but also to appropriate assessment and management, especially among disadvantaged groups. PMID- 17137436 TI - Revisiting the metabolic syndrome. AB - Metabolic syndrome (MS) refers to the clustering of cardiometabolic risk factors including abdominal obesity, hyperglycaemia, dyslipidaemia and elevated blood pressure - that are thought to be linked to insulin resistance. MS is associated with increased risk of cardiovascular disease and type 2 diabetes. MS is common, affecting a quarter to a third of adults, and its prevalence is rising, in parallel with increasing obesity and population ageing. Operational definitions of MS have been proposed by the World Health Organization and the National Cholesterol Education Program. Recently, the International Diabetes Federation proposed a global definition that emphasised the importance of central adiposity. In cardiovascular risk assessment, MS encapsulates the contribution of non traditional risk factors and provides a clinically useful framework for early identification of people at increased long-term risk. It should be used in conjunction with standard algorithms based on conventional risk factors, which better predict short-term risk. Management of MS should emphasise lifestyle interventions (eg, physical activity, healthy diet and weight reduction) to reduce long-term risk of cardiovascular disease and diabetes. Those at increased short-term risk should also have individual risk factors treated according to established guidelines. PMID- 17137437 TI - Men who have sex with men (MSM): how much to assume and what to ask? AB - In Australia, about 150,000 men aged 16-59 years identify as gay or bisexual, while a similar number identify as heterosexual but have some history of same-sex sexual contact. Pitts, Couch and Smith advise that the clinical implications for these men include more than sexual health concerns. They suggest several consultation skills that can help doctors to recognise these men and better meet their needs. PMID- 17137438 TI - Mental health initiatives for veterans and serving personnel. PMID- 17137439 TI - Is there a case for differential treatment of young men and women? PMID- 17137440 TI - Shifting paradigms: a social-determinants approach to solving problems in men's health policy and practice. AB - The lack of an evidence base for formulating men's health policies means existing programs and practices for men are influenced by prevailing cultural norms concerning men or habitual health service attitudes towards them. Factors impeding the development of an effective health policy for men include a preoccupation with limited clinical perspectives (an emphasis on the prostate and erectile dysfunction) and a common assumption that all health problems in men are a result of "masculinity" and "men behaving badly". Viewing men's health in terms of gender and health and the socially constructed differences between men and women is important, but does not provide all the perspectives required for meeting men's health needs. A "social determinants of health" approach to men's health would help Australia and Australian medical practitioners move away from policies and practices that perpetuate negative views of men and ignore the complexity of their health problems. The result would be a more evidence-based approach to men's health policy, and the likelihood of improved health outcomes. PMID- 17137441 TI - What is it with men's health?" Men, their health and the system: a personal perspective. PMID- 17137442 TI - Addressing depression and anxiety among new fathers. AB - Fathers may be unintentionally marginalised by perinatal health services and by the maternal focus of social practices surrounding new babies. There is increasing recognition that a fathers' depression and anxiety in the perinatal period can have serious consequences for his family. Health services could better support new fathers by providing them with information on parenting from a father's perspective, or by running father-specific sessions as part of routine antenatal care programs. PMID- 17137443 TI - Growing up brown in a white-shirted time. PMID- 17137444 TI - Aboriginal and Torres Strait Islander male health, wellbeing and leadership. PMID- 17137445 TI - Gay with the experience of disability: the ideal man! PMID- 17137446 TI - What boys need: a sense of "belonging". PMID- 17137447 TI - Woody's story: fighting prostate cancer. PMID- 17137448 TI - Cervical cancer prevention: the saga goes on, but so much has changed! PMID- 17137449 TI - The limits of perinatal viability: grappling with the "grey zone". PMID- 17137450 TI - Inequity in rural cancer survival in Australia is not an insurmountable problem. PMID- 17137451 TI - Cervical cancer in Australia and the United Kingdom: comparison of screening policy and uptake, and cancer incidence and mortality. AB - OBJECTIVE: To compare cervical screening policy, screening uptake, and changes in cervical cancer incidence and mortality between Australia and the United Kingdom. DESIGN: Analysis of screening registry data and national cancer statistics. SETTING: In Australia, organised cervical screening was initiated in 1991 for sexually active women aged 18-69 years, with a recommended 2-yearly interval. In the UK, organised screening began in 1988 for women aged 20-64 years, with a recommended 3-yearly interval in most regions. RESULTS: Estimated lifetime screening participation rates in 2001 were similar in the two countries, at 88% in Australia and 90% in the UK. For women who were screened and had a negative result, the median time to the next screen was 27 months in Australia and 38 months in the UK. At 39 months, equivalent proportions (74%) had been re-screened in the two countries, and by 60 months the re-screened proportions were 81% in Australia and 94% in the UK. From 1991-1993 to 1998-2000, the incidence of cervical cancer in women aged 20-69 years fell by 33% in Australia and 33% in the UK, and mortality from cervical cancer fell by 36% in both countries. CONCLUSIONS: After the introduction of organised screening, similar reductions in cervical cancer incidence and mortality were achieved in Australia and the UK. Therefore, the 2-yearly screening policy in Australia and the predominantly 3 yearly screening policy in the UK appear to have been of broadly similar effectiveness. PMID- 17137452 TI - Self-reported adherence with medication and cardiovascular disease outcomes in the Second Australian National Blood Pressure Study (ANBP2). AB - OBJECTIVE: To investigate whether responses to a previously validated four-item medication adherence questionnaire were associated with adverse cardiovascular events. DESIGN: Survey conducted among a cohort of participants in the Second Australian National Blood Pressure Study. SETTING: Australian general practice. PARTICIPANTS: 4039 older people with hypertension. MAIN OUTCOME MEASURES: All major cardiovascular events or death; first specific cardiovascular event. RESULTS: Subjects who adhered to their medication regimen (compared with non adherent subjects) were significantly less likely to experience a first cardiovascular event or a first non-fatal cardiovascular event (hazard ratio [HR] for both, 0.81; 95% CI, 0.67-0.98; P = 0.03); a fatal other cardiovascular event (HR, 0.68; 95% CI, 0.48-0.99; P = 0.04); or a first occurrence of heart failure (HR, 0.58; 95% CI, 0.37-0.90; P = 0.02). Those who answered yes to "Did you ever forget to take your medication?" were significantly more likely to experience a cardiovascular event or death (HR, 1.28; 95% CI, 1.04-1.57; P = 0.02); a first cardiovascular event or death (HR, 1.31; 95% CI, 1.07-1.60; P = 0.01); a first cardiovascular event (HR, 1.34; 95% CI, 1.09-1.65; P = 0.01); or a first non fatal cardiovascular event (HR, 1.35; 95% CI, 1.09-1.66; P = 0.01). Those who answered yes to "Sometimes, if you felt worse when you took your medicine, did you stop taking it?" were significantly more likely to experience a first occurrence of heart failure (HR, 2.06; 95% CI, 1.16-3.64; P = 0.01). CONCLUSIONS: Subjects who adhered to their medication regimen were less likely to experience major cardiovascular events or death. The question relating to forgetting to take medication identified non-adherent subjects likely to experience a cardiovascular event or death. Clinicians could use this question to identify patients with hypertension who are likely to benefit from medication adherence strategies. PMID- 17137453 TI - Participation in cervical screening by Indigenous women in the Northern Territory: a longitudinal study. AB - OBJECTIVE: To investigate the effectiveness of the Northern Territory Women's Cancer Prevention Program in improving cervical screening participation for Indigenous women. DESIGN: Descriptive longitudinal period prevalence study. PARTICIPANTS: All NT resident women aged 20-69 years who had at least one Pap smear recorded on the NT Pap Smear Register between 1997 and 2004. MAIN OUTCOME MEASURES: Indirectly estimated percentage of NT Indigenous women in rural and remote areas with a predominantly Indigenous population (accounting for 55% of the NT Indigenous population) who participated in screening, in biennial periods between 1997 and 2004. Participation by all eligible NT women (both Indigenous and non-Indigenous) is also reported by region for the same period. RESULTS: In 1997-1998, estimated participation for Indigenous women was about half the national rate (33.9% [95% CI, 32.6%-35.2%] v 63.9% [95% CI, 63.8%-63.9%]). Participation increased to 44.0% (95% CI, 42.7%-45.4%) in 1999-2000, and changed little thereafter; participation was higher in the Top End compared with Central Australia, and varied from 16.6% to 75.0% between remote areas. Participation rates for all women living in rural/remote regions were lower than those in urban regions. CONCLUSIONS: Recruitment of Indigenous women for cervical screening has improved since 1999. This may have partly contributed to the fall in their cervical cancer incidence and mortality in recent years. Although in most areas Indigenous participation is lower than national levels, in one area it was considerably higher. Improvements can be achieved by learning from these communities, to further close the gap in morbidity and mortality between Indigenous and non-Indigenous women. PMID- 17137454 TI - Perinatal care at the borderlines of viability: a consensus statement based on a NSW and ACT consensus workshop. AB - Perinatal care at the borderlines of viability demands a delicate balance between parents' wishes and autonomy, biological feasibility, clinicians' responsibilities and expectations, and the prospects of an acceptable long-term outcome - coupled with a tolerable margin of uncertainty. A multi-professional workshop with consumer involvement was held in February 2005 to agree on management of this issue in New South Wales and the Australian Capital Territory. Participants discussed and formulated consensus statements after an extensive consultation process. Consensus was reached that the "grey zone" is between 23 weeks' and 25 weeks and 6 days' gestation. While there is an increasing obligation to treat with increasing length of gestation, it is acceptable medical practice not to initiate intensive care during this period if parents so wish, after appropriate counselling. Poor condition at birth and the presence of serious congenital anomalies have an important influence on any decision not to initiate intensive care within the grey zone. Women at high risk of imminent delivery within the grey zone should receive appropriate and skilled counselling with the most relevant up-to-date outcome information. Management plans can thus be made before birth. Information should be simple, factual and consistent. The consensus statements developed will provide a framework to assist parents and clinicians in communication, decision making and managing these challenging situations. PMID- 17137455 TI - Risk of serious NSAID-related gastrointestinal events during long-term exposure: a systematic review. AB - OBJECTIVE: Exposure to non-steroidal anti-inflammatory drugs (NSAIDs) is associated with increased risk of serious gastrointestinal (GI) events compared with non-exposure. We investigated whether that risk is sustained over time. DATA SOURCES: Cochrane Controlled Trials Register (to 2002); MEDLINE, EMBASE, Derwent Drug File and Current Contents (1999-2002); manual searching of reviews (1999 2002). STUDY SELECTION: From 479 search results reviewed and 221 articles retrieved, seven studies of patients exposed to prescription non-selective NSAIDs for more than 6 months and reporting time-dependent serious GI event rates were selected for quantitative data synthesis. These were stratified into two groups by study design. DATA EXTRACTION: Incidence of GI events and number of patients at specific time points were extracted. DATA SYNTHESIS: Meta-regression analyses were performed. Change in risk was evaluated by testing whether the slope of the regression line declined over time. Four randomised controlled trials (RCTs) provided evaluable data from five NSAID arms (aspirin, naproxen, two ibuprofen arms, and diclofenac). When the RCT data were combined, a small significant decline in annualised risk was seen: - 0.005% (95% CI, - 0.008% to - 0.001%) per month. Sensitivity analyses were conducted because there was disparity within the RCT data. The pooled estimate from three cohort studies showed no significant decline in annualised risk over periods up to 2 years: - 0.003% (95% CI, - 0.008% to 0.003%) per month. CONCLUSIONS: Small decreases in risk over time were observed; these were of negligible clinical importance. For patients who need long-term (> 6 months) treatment, precautionary measures should be considered to reduce the net probability of serious GI events over the anticipated treatment duration. The effect of intermittent versus regular daily therapy on long-term risk needs further investigation. PMID- 17137456 TI - Genetic counselling for psychiatric disorders. AB - Family, adoption and twin studies demonstrate that many adult psychiatric disorders, including schizophrenia, major depression and bipolar disorder, have a clear genetic component. The aetiology of psychiatric disorders is a complex combination of both genetic and environmental components. While potential susceptibility genes for psychiatric disorders have been identified, interaction with the environment is a crucial component in disease development. Pharmacogenetics and genetic testing have the potential to play key roles in the future of clinical psychiatry. At present, an increased risk of psychiatric disorders can be identified through a detailed family history. The empirical risk of developing a disorder has been determined for many psychiatric disorders and can be used as a general guide. Genetic counselling can extend and enhance patient care by providing information to patients about the complexities of inheriting psychiatric disorders and the associated risks of recurrence. The genetic counselling process can facilitate informed decision making, alleviate misconceptions and reduce stigma through an improved understanding of the genetic cause of psychiatric disorders, and offer support to patients and their families. PMID- 17137457 TI - Tackling the burden of injury in Australasia: developing a binational trauma registry. AB - Existing trauma registries in Australia and New Zealand play an important role in monitoring the management of injured patients. Over the past decade, such monitoring has been translated into changes in clinical processes and practices. Monitoring and changes have been ad hoc, as there are currently no Australasian benchmarks for "optimal" injury management. A binational trauma registry is urgently needed to benchmark injury management to improve outcomes for injured patients. PMID- 17137458 TI - Loss of an eye in a baby from keratitis initially managed as conjunctivitis. PMID- 17137459 TI - 5. Allergy and the skin: eczema and chronic urticaria. AB - Eczema is common, occurring in 15%-20% of infants and young children. For some infants it can be a severe chronic illness with a major impact on the child's general health and on the family. A minority of children will continue to have eczema as adults. The exact cause of eczema is not clear, but precipitating or aggravating factors may include food allergens (most commonly, egg) or environmental allergens/irritants, climatic conditions, stress and genetic predisposition. Management of eczema consists of education; avoidance of triggers and allergens; liberal use of emollients or topical steroids to control inflammation; use of antihistamines to reduce itch; and treatment of infection if present. Treatment with systemic agents may be required in severe cases, but must be supervised by an immunologist. Urticaria ("hives") may affect up to a quarter of people at some time in their lives. Acute urticaria is more common in children, while chronic urticaria is more common in adults. Chronic urticaria is not life-threatening, but the associated pruritus and unsightly weals can cause patients much distress and significantly affect their daily lives. Angioedema coexists with urticaria in about 50% of patients. It typically affects the lips, eyelids, palms, soles and genitalia. Management of urticaria is through education; avoidance of triggers and allergens (where relevant); use of antihistamines to reduce itch; and short-term use of corticosteroids when antihistamine therapy is ineffective. Referral is indicated for patients with resistant disease. PMID- 17137460 TI - Vernal keratoconjunctivitis. PMID- 17137461 TI - Guidelines for the management of acute coronary syndromes 2006. PMID- 17137462 TI - The quality of national data on injuries requiring hospitalisation. PMID- 17137463 TI - Policy lags behind reality on antenatal HIV screening. PMID- 17137464 TI - Should clinical software be regulated? PMID- 17137465 TI - A hexanucleotide selected for increased cellular uptake in cis contains a highly active CpG-motif in human B cells and primary peripheral blood mononuclear cells. AB - The relationship between immunostimulation of human B cells by cytosine-phosphate guanosine (CpG) -containing oligonucleotides and their physical cellular uptake is of mechanistic interest and a prerequisite for rational improvements of the therapeutic potential of CpG-harbouring oligonucleotides. Here, a combinatorial approach was used to identify nucleotide sequence motifs that facilitate increased cellular uptake in mammalian cells. Oligonucleotides harbouring the selected hexanucleotide TCGTGT in cis show increased cellular uptake. This motif contains a CpG dinucleotide within a sequence context that shows a very strong CpG-specific stimulatory activity on human B cells. Here we describe the influence of concentration, length and sequence position of the unmethylated CpG dinucleotide on immunostimulation. A comparison between phosphorothioate derivatives and unmodified TCGTGT-containing oligonucleotides strongly indicates a great CpG-specificity for the unmodified CpG-harbouring oligonucleotides but not for the phosphorothioate versions. This work describes a link between the physical cellular uptake of naked oligonucleotides harbouring the selected cellular uptake motif TCGTGT, its strong CpG-specific stimulation of human B cells and its relationship with the sequence context of CpG and its cellular uptake. PMID- 17137466 TI - Dopaminergic deficiency in mice with reduced levels of the dual-specificity tyrosine-phosphorylated and regulated kinase 1A, Dyrk1A(+/-). AB - The dual-specificity tyrosine-phosphorylated and regulated kinase 1A (DYRK1A) gene encodes a protein kinase known to play a critical role in neurodevelopment. Mice with one functional copy of Dyrk1A (Dyrk1A(+/-)) display a marked hypoactivity and altered gait dynamics in basal conditions and in novel environments. Dopamine (DA) is a key neurotransmitter in motor behavior and genetic deletion of certain genes directly related to the dopaminergic system has a strong impact on motor activity. We have studied the effects of reduced Dyrk1A expression on the function of the nigrostriatal dopaminergic system. To characterize the dopaminergic system in DYRK1A(+/-) mice, we have used behavioral, pharmacological, histological, neurochemical and neuroimaging (microPET) techniques in a multidisciplinary approach. Dyrk1A(+/-) mice exhibited decreased striatal DA levels, reduced number of DA neurons in the substantia nigra pars compacta, as well as altered behavioral responses to dopaminergic agents. Moreover, microdialysis experiments revealed attenuated striatal DA release and positron emission tomography scan display reduced forebrain activation when challenged with amphetamine, in Dyrk1A(+/-) compared with wild type mice. These data indicate that Dyrk1A is essential for a proper function of nigrostriatal dopaminergic neurons and suggest that Dyrk1A(+/-) mice can be used to study the pathogenesis of motor disorders involving dopaminergic dysfunction. PMID- 17137467 TI - Bone regeneration in dehiscence-type defects at chemically modified (SLActive) and conventional SLA titanium implants: a pilot study in dogs. AB - OBJECTIVES: The aim of the present study was to evaluate bone regeneration in dehiscence-type defects at titanium implants with chemically modified (mod) and conventional sand-blasted/acid-etched (SLA) surfaces. MATERIAL AND METHODS: Standardized buccal dehiscence defects (height: 3 mm, width: 3 mm) were surgically created following implant site preparation in both the upper and lower jaws of four beagle dogs. modSLA and SLA implants were inserted bilaterally according to a split-mouth design. The animals were sacrificed after 2 and 12 weeks (n=2 animals each). Dissected blocks were processed for histomorphometrical analysis: defect length, new bone height (NBH), percent linear fill (PLF), percent of bone-to-implant contact (BIC-D) and area of new bone fill (BF). RESULTS: Wound healing at SLA implants was predominantly characterized by the formation of a dense connective tissue at 2 and 12 weeks, without significant increases in mean NBH, PLF, BIC-D or BF values. In contrast, modSLA implants exhibited a complete defect fill at 12 weeks following implant placement. In particular, histomorphometrical analysis revealed the following mean values at 12 weeks: NBH (3.2+/-0.3 mm), PLF (98%), BIC-D (82%) and BF (2.3+/-0.4 mm(2)). CONCLUSION: Within the limits of the present study, it was concluded that modSLA titanium surfaces may promote bone regeneration in acute-type buccal dehiscence defects at submerged implants. PMID- 17137468 TI - Does periodontal care improve glycemic control? The Department of Veterans Affairs Dental Diabetes Study. AB - OBJECTIVES: Report results of a randomized-clinical trial of the efficacy of periodontal care in the improvement of glycemic control in 165 veterans with poorly controlled diabetes over 4 months. METHODS: Outcomes were change in Haemoglobin A1c (HbA1c) in the Early Treatment versus untreated (Usual Care) groups and percent of participants with decreases in HbA1c. Analyses included simple/multiple variable linear/logistic regressions, adjusted for baseline HbA1c, age, and duration of diabetes. RESULTS: Unadjusted analyses showed no differences between groups. After adjustment for baseline HbA1c, age, and diabetes duration, the mean absolute HbA1c change in the Early Treatment group was -0.65% versus -0.51% in the Usual Care group (p=0.47). Adjusted odds for improvement by 0.5% in the Early Treatment group was 1.67 (95% confidence interval: 0.84, 3.34, p=0.14). Usual Care subjects were twice as likely to increase insulin from baseline to 4 months (20% versus 11%, p=0.12) and less likely to decrease insulin (1% versus 6%, p=0.21) than Early Treatment subjects. Among insulin users at baseline, more increased insulin in the Usual Care group (40% versus 21%, p=0.06). CONCLUSIONS: No significant benefit was found for periodontal therapy after 4 months in this study; trends in some results were in favour of periodontal treatment. PMID- 17137469 TI - A three-year prospective study of adult subjects with gingivitis. I: clinical periodontal parameters. AB - AIM: The objective of this study was to monitor prospectively clinical parameters in subjects without signs of destructive periodontal disease who were involved in a primary prevention programme, and to determine the changes that occurred between yearly examinations over a 3-year period. MATERIAL AND METHODS: One hundred and twenty-six subjects aged at least 20 years with a maximum of two tooth sites with probing pocket depth (PPD)>4 mm and no proximal sites with clinical attachment loss participated in the study. Primary prevention was provided at baseline of the study and then every 6 months. Plaque, bleeding on probing (BoP) and PPD were scored at baseline, 1, 2 and 3 years. RESULTS: There were no significant changes in the plaque score over the 3 years. After year 1, the BoP score was significantly improved with 5.6%, while no further improvement in BoP was found at years 2 and 3. The mean PPD decreased from 2.3 to 2.1 mm over the 3 years (p<0.05). CONCLUSION: Although some individuals exhibiting minor signs of periodontal pathology may have benefited from the primary prevention, the overall clinical improvement was limited for such subjects in the present 3 year study. PMID- 17137470 TI - Long-term effects of hemostatic variables on fatal coronary heart disease: 30 year results from the first prospective Northwick Park Heart Study (NPHS-I). AB - BACKGROUND: The long-term associations of established risk factors for coronary heart disease (CHD), for example cholesterol, are well known, but not for the less familiar hemostatic variables. OBJECTIVES: To establish whether associations between hemostatic variables and CHD first identified nearly three decades ago have persisted long-term. METHODS: The first Northwick Park Heart Study (NPHS-I) recruited 2167 white men and 941 white women, average age at entry 48 years, on whom measures of factor (F) VII activity (VIIc) and plasma fibrinogen were carried out, both at entry and at follow-up approximately 6 years later. RESULTS: During a median follow-up of 29 years, 231 male and 36 female CHD deaths were recorded from notifications by the Office for National Statistics. VIIc at recruitment was significantly related to CHD mortality, corrected rate ratio, RR, per 1 SD increase 1.56 (95% CI 1.29, 1.88) in men and RR 1.78 (95% CI 1.17, 2.72) in women. Recruitment fibrinogen was also strongly related to CHD mortality in men, RR 1.63 (95% CI 1.33, 1.99) but not in women, RR 0.75 (95% CI 0.40, 1.43). The associations persisted after controlling for confounders and were confirmed using 6-year follow-up measurements and in analyses omitting deaths within 10 years of recruitment. CONCLUSIONS: The hemostatic system contributes to CHD mortality, and its effect is stable over time. For VIIc, the effect was similar in men and women, while for fibrinogen it appeared to be present only in men. PMID- 17137471 TI - Defective platelet responsiveness to thrombin and protease-activated receptors agonists in a novel case of gray platelet syndrome: correlation between the platelet defect and the alpha-granule content in the patient and four relatives. AB - BACKGROUND: We report a novel case of gray platelet syndrome (GPS). A 14-year-old boy had bleeding diathesis, mild thrombocytopenia, giant platelets with severe defect of alpha-granule secretory proteins, myelofibrosis and splenomegaly. METHODS AND RESULTS: Platelet function studies showed a marked reduction of aggregation and Ca(2+) mobilization by thrombin, protease-activated receptor 1 (PAR1)-activating peptide (AP) and PAR4-AP, PAR1 expression at 55% of normal levels, and a more than two hundred fold reduction of in vitro whole-blood thromboxane B(2) (TXB(2)) production. Sequencing of coding regions of the PAR1 gene failed to show abnormalities. This patient was initially classified as a sporadic case of GPS, as electron microscopy failed to identify giant platelets and/or alpha-granule deficiency in his relatives. However, further studies on the father and three other relatives showed a relative lack of platelet alpha-granule proteins by immunofluorescence microscopy, a defective platelet response to PAR4 AP, and severely reduced in vitro whole-blood TXB(2) production. On this basis, we suggest that in this family, GPS was transmitted in a dominant fashion with highly variable penetrance. CONCLUSIONS: Our study suggests that current diagnostic criteria fail to identify some patients with a mild GPS phenotype and that such patients might be identified by the methods cited above. It also better characterizes the pathogenesis of defective platelet responses to thrombin, and raises interesting questions on the correlation between abnormal PAR function and the lack of alpha-granule content in GPS. PMID- 17137472 TI - Risk of excessive bleeding associated with marginally low von Willebrand factor and mild platelet dysfunction. AB - BACKGROUND: Bleeding symptoms are so commonly reported that it is not known whether they associate causally or coincidentally with mild but measurable primary hemostatic defects. OBJECTIVES/PATIENTS/METHODS: In order to evaluate if the mild primary hemostatic defects are truly causative of increased bleeding symptoms, we surveyed a population of healthy teenagers for bleeding symptoms. Using a case-control approach, we then estimated the risk of excessive bleeding associated with low von Willebrand factor (defined as VWF below the 5th percentile of a normal reference population), and with mild platelet dysfunction [PD, defined as concurrent reduced platelet aggregation responses to two agonists (adenosine diphosphate and epinephrine)]. RESULTS: Excessive bleeding was present in 63 out of 809 teenagers (7.8%). Among the 49 cases who were tested for VWF, low values by three measures were more commonly present than in 166 controls, specifically, ristocetin cofactor (RCo) activity [20.4% vs. 5.4%, odds ratio (OR) 4.5], collagen binding (14.3% vs. 4.2%, OR 3.8), and antigen level (20.4% vs. 6.0%, OR 4.0). The low RCo values ranged from 35 to 45 U dL(-1) except for a single case with 26 U dL(-1). Of the 47 teenagers with excessive bleeding who underwent platelet aggregation studies, reduced responses were more common than in controls (12.8% vs. 4.4%, OR 3.2). Twenty-nine per cent of cases with excessive bleeding had either low RCo or PD. CONCLUSION: Almost one in three teenagers who report excessive bleeding is likely to have a measurable hemostatic disturbance manifested either by marginally low VWF (by three measures) or by mild PD. PMID- 17137473 TI - Nodular prurigo: metabolic diseases are a common association. PMID- 17137474 TI - The effect of narrowband ultraviolet B on the expression of matrix metalloproteinase-1, transforming growth factor-beta1 and type I collagen in human skin fibroblasts. AB - BACKGROUND: Ultraviolet (UV) irradiation induces chronic skin diseases, such as skin cancer and photoageing, and the mechanisms of this skin damage are associated with the upregulation of matrix metalloproteinases (MMPs) and decreased collagen synthesis. Narrowband ultraviolet B (NB-UVB) radiation is a relatively new treatment modality for vitiligo and psoriasis. However, the mechanism of NB-UVB action on photoageing is not completely understood. Aims. We investigated the effects of NB-UVB on the expression of MMP-1, transforming growth factor (TGF)-beta1 and type I collagen in cultured human skin fibroblasts. METHODS: Cultured human fibroblasts were irradiated with either NB-UVB (50-800 mJ/cm(2)) or broadband UVB (BB-UVB; 25 mJ/cm(2)). The expression of MMP-1, TGF beta1 and type I collagen mRNA was determined by reverse-transcription PCR. Expression of MMP-1 and TGF-beta1 protein was determined by ELISA and that of type I collagen by Western blotting. RESULTS: NB-UVB induced the expression of MMP-1 and reduced the expression of TGF-beta1 and type I collagen at the mRNA and protein levels in a dose-dependent manner. The expression of type I collagen protein decreased more after irradiation with 25 mJ/cm(2) of BB-UVB than 400 mJ/cm(2) of NB-UVB. CONCLUSIONS: This study indicates that NB-UVB irradiation reduces type I collagen synthesis in human skin fibroblasts by inhibiting TGF beta1 expression and stimulating the release of MMP-1. It also suggested that the photoageing-related effects of NB-UVB are weaker than those of BB-UVB in vitro. PMID- 17137475 TI - Long-term clearance of patch-stage mycosis fungoides with the 308-nm laser. AB - In order to investigate the effectiveness of the 308-nm xenon chloride laser in the treatment of patch-stage mycosis fungoides that is refractory to topical steroids, we conducted a prospective, histologically controlled, open-label pilot study. Eight patients with stage 1A or 1B cutaneous T-cell lymphoma that was unresponsive to application of potent topical steroids received 20 treatments with the laser over 10 weeks and were then followed up for a minimum of 30 months. Three patients had a complete clinical and histological response and remained free of disease after 30 months A further three patients had complete clinical and histological clearance of disease, but experienced relapse within the 30 months follow-up period. One patient withdrew from the study and one had a partial response. Further research to establish the role of the 308-nm laser in the treatment of MF appears justified. PMID- 17137476 TI - Disseminated superficial actinic porokeratosis of the scalp. PMID- 17137477 TI - Interobserver reliability of the Nail Psoriasis Severity Index. AB - BACKGROUND: Because the Psoriasis Area and Severity Index (PASI) does not consider the severity of nail disease, a scale that assesses the extent of involvement of psoriatic nails is needed. A new grading system, the Nail Psoriasis Severity Index (NAPSI) has been proposed. AIMS: The purpose of this study was to assess the interobserver reliability of NAPSI. Methods. The nail features of 25 patients with psoriasis with nail involvement were evaluated and graded by three dermatologists for total NAPSI scores and nail scores. The quadrants of all nails were examined for the presence of matrix and bed features. Total NAPSI score (0-160) of patients and nail score (0-32) of the individual nails were calculated. Interobserver reliability assessments were performed by computing intraclass correlation coefficients (ICC; two-way mixed model, consistency definition). RESULTS: The ICC((3,1)) results for total NAPSI score and nail score were found to be 0.781 and 0.649, respectively. The ICC((3,1)) for nail-bed and nail-matrix features were 0.869 and 0.584, respectively, in the total NAPSI scoring system, and 0.705 and 0.603, respectively, in the nail scoring system. CONCLUSION: Moderate to good agreement of scoring with the NAPSI was determined among the observers in this study. Our results suggest that scoring for nail-bed features seems to be more reliable than scoring for nail matrix features. PMID- 17137478 TI - Epidermoid cysts mimicking recurrence of superficial basal cell carcinoma following photodynamic therapy. PMID- 17137479 TI - Vasculopathic skin lesions following epilation, leading to a discovery of hepatitis C virus infection. PMID- 17137480 TI - Pyoderma gangrenosum on varicella lesions. PMID- 17137481 TI - Erythema multiforme associated with resolving molluscum contagiosum. PMID- 17137482 TI - A patch of alopecia in a 70-year-old woman. PMID- 17137483 TI - Bier's white spots associated with scleroderma renal crisis. AB - Bier's spots are asymptomatic and permanent white macules, standing on a cyanotic background. These spots are associated with venous stasis and usually appear in physiological conditions. We report the case of a 47-year-old woman with a 2-year history of systemic scleroderma, who developed Bier's white spots associated with a vascular and renal crisis. Interestingly, these spots appeared with the crisis, and disappeared with the treatment and resolution of the crisis. They are probably a result of anatomical and functional damage to the small vessels of the skin. The pathophysiology in the skin is probably the same as that which happens in renal vessels during scleroderma renal crisis. PMID- 17137484 TI - Effects of moisturization on epidermal homeostasis and differentiation. AB - Moisturizers are commonly used for routine skin care. This study assessed the effects of a moisturizer on barrier function, epidermal architecture, keratinocyte proliferation, and physiological regulation of the epidermis in photoaged but otherwise normal skin. Fifteen women with moderately photoaged forearms were treated twice a day for 4 weeks with a moisturizer containing dimethicone and glycerine. Baseline and post-treatment transepidermal water loss (TEWL) and ipsilateral forearm biopsies were obtained. Epidermal thickness, melanin levels, keratinocyte proliferation, and expression of keratins were evaluated. Induction of keratins 6 and 16, commonly associated with keratinocyte proliferation and wound healing, was observed. Epidermal thickness increased by 0.019 mm (P = 0.005), barrier function improved (TEWL decreased by 13%) and melanin intensity decreased (P = 0.004). Even nonxerotic, photoaged skin may appear younger, benefiting structurally and functionally from routine use of moisturizers containing dimethicone and glycerine. PMID- 17137485 TI - Complete ophthalmoplegia after herpes zoster. AB - Motor loss caused by herpes zoster is infrequent, and only a few studies have focused on ocular motor paralysis in ophthalmic herpes zoster. We report a case of complete ophthalmoplegia resulting from ophthalmic herpes zoster. A 69-year old man presented with complete left-side ptosis with total ophthalmoplegia 7 days after the onset of left ophthalmic herpes zoster. The patient was treated with aciclovir and prednisolone. Five months later, the ptosis had resolved and the extraocular motility had almost returned to normal. PMID- 17137486 TI - Pathergy reaction in different body areas in Behcet's disease. AB - Behcet's disease (BD) is a chronic inflammatory multisystemic disorder characterized mainly by vasculitis. Although the pathergy reaction (PR) has been accepted as one of the major criteria in this disease, there is a lack of a standardized method to measure PR. The aim of this study was to evaluate results of PR in different body areas and the effect on the positive rate of PR of application of numerous needle pricks. The pathergy test was performed using simultaneous 16 needle pricks with 20G disposable needles on different body areas of 56 patients with BD, 49 patients with recurrent aphthous stomatitis (RAS) and 17 healthy subjects. No positive PR was found in healthy controls, but was observed in nine patients with RAS (18.3%). It was detected in 48 of 56 patients with BD, a positive rate of 85.7%. The site most frequently positive for PR was the forearm, while the least frequent was the abdomen. Multiple needle pricks could play an important role in increasing the positive rate of PR in BD, and it may be important in the investigation of aetiological factors in RAS. PMID- 17137487 TI - Photographic artefact resembling melanocytic naevi. PMID- 17137488 TI - A painful cutaneous eruption in a pregnant woman. PMID- 17137489 TI - Influence of mycotoxin zearalenone and its derivatives (alpha and beta zearalenol) on apoptosis and proliferation of cultured granulosa cells from equine ovaries. AB - BACKGROUND: The mycotoxin zearalenone (ZEA) and its derivatives, alpha and beta zearalenol (alpha and beta-ZOL), synthesized by genera Fusarium, often occur as contaminants in cereal grains and animal feeds. The importance of ZEA on reproductive disorders is well known in domestic animals species, particularly in swine and cattle. In the horse, limited data are available to date on the influence of dietary exposure to ZEA on reproductive health and on its in vitro effects on reproductive cells. The aim of this study was to evaluate the effects of ZEA and its derivatives, alpha and beta-ZOL, on granulosa cells (GCs) from the ovaries of cycling mares. METHODS: The cell proliferation was evaluated by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test after 3 days exposure at different concentrations of ZEA and its derivatives (from 1 x 10-7 to 0.1 microM). The apoptosis induction was evaluated after 1 day exposure, by DNA analysis using flow cytometry. RESULTS: An increase in cell proliferation with respect to the control was observed in the presence of ZEA at 1 x 10-3 and 1 x 10-4 microM and apoptosis was induced by all mycotoxins at different concentrations. CONCLUSION: The simultaneous presence of apoptosis and proliferation in GC cultures treated with zearalenones could indicate that these mycotoxins could be effective in inducing follicular atresia. These effects of zearalenones may result from both direct interaction with oestrogen-receptors as well as interaction with the enzymes 3alpha (beta)-hydroxysteroid dehydrogenase (HSD), involved in the synthesis and metabolism of endogenous steroid hormones. These cellular disturbances, described for the first time in equine GCs cultured in vitro, could be hypothesized as referred to reproductive failures of unknown ethiology in the mare. PMID- 17137490 TI - Inferring topology from clustering coefficients in protein-protein interaction networks. AB - BACKGROUND: Although protein-protein interaction networks determined with high throughput methods are incomplete, they are commonly used to infer the topology of the complete interactome. These partial networks often show a scale-free behavior with only a few proteins having many and the majority having only a few connections. Recently, the possibility was suggested that this scale-free nature may not actually reflect the topology of the complete interactome but could also be due to the error proneness and incompleteness of large-scale experiments. RESULTS: In this paper, we investigate the effect of limited sampling on average clustering coefficients and how this can help to more confidently exclude possible topology models for the complete interactome. Both analytical and simulation results for different network topologies indicate that partial sampling alone lowers the clustering coefficient of all networks tremendously. Furthermore, we extend the original sampling model by also including spurious interactions via a preferential attachment process. Simulations of this extended model show that the effect of wrong interactions on clustering coefficients depends strongly on the skewness of the original topology and on the degree of randomness of clustering coefficients in the corresponding networks. CONCLUSION: Our findings suggest that the complete interactome is either highly skewed such as e.g. in scale-free networks or is at least highly clustered. Although the correct topology of the interactome may not be inferred beyond any reasonable doubt from the interaction networks available, a number of topologies can nevertheless be excluded with high confidence. PMID- 17137491 TI - Increased endothelin-1 and diminished nitric oxide levels in blister fluids of patients with intermediate cold type complex regional pain syndrome type 1. AB - BACKGROUND: In complex regional pain syndrome type 1 (CRPS1) pro-inflammatory mediators and vascular changes play an important role in the sustained development and outcome of the disease. The aim of this study was to determine the involvement of vasoactive substances endothelin-1 (ET-1) and nitric oxide (NO) during early chronic CRPS1. METHODS: Included were 29 patients with CRPS 1 who were diagnosed during the acute stage of their disease and observed during follow-up visits. Disease activity and impairment were determined and artificial suction blisters were made on the CRPS1 and the contralateral extremities for measurements of IL-6, TNF-alpha, ET-1 and nitrate/nitrite (NOx). RESULTS: The levels of IL-6, TNF-alpha and ET-1 in blister fluid in the CRPS1 extremity versus the contralateral extremity were significantly increased and correlated with each other, whereas NOx levels were decreased. CONCLUSION: The NOx/ET-1 ratio appears to be disturbed in the intermediate stage of CRPS, resulting in vasoconstriction and consequently in a diminished tissue blood distribution. PMID- 17137492 TI - A type I and type II microsatellite linkage map of rainbow trout (Oncorhynchus mykiss) with presumptive coverage of all chromosome arms. AB - BACKGROUND: The development of large genomic resources has become a prerequisite to elucidate the wide-scale evolution of genomes and the molecular basis of complex traits. Linkage maps represent a first level of integration and utilization of such resources and the primary framework for molecular analyses of quantitative traits. Previously published linkage maps have already outlined the main peculiarities of the rainbow trout meiosis and a correspondance between linkage groups and chromosome arms has been recently established using fluorescent in situ hybridization. The number of chromosome arms which were covered by these maps remained unknown. RESULTS: We report an updated linkage map based on segregation analysis of more than nine hundred microsatellite markers in two doubled haploid gynogenetic lines. These markers segregated into 31 linkage groups spanning an approximate total map length of 2750 cM. Centromeres were mapped for all the linkage groups using meiogenetic lines. For each of the 31 linkage groups, the meta or acrocentric structure infered from centromere mapping was identical with those recently found with fluorescent in situ hybridization results. The present map is therefore assumed to cover the 52 chromosome arms which constitute the rainbow trout karyotype. Our data confirm the occurrence of a high interference level in this species. Homeologous regions were identified in eleven linkage groups, reflecting the tetraploid nature of the salmonid genome. The data supported the assumption that gene orders are conserved between duplicated groups and that each group is located on a single chromosome arm. Overall, a high congruence with already published rainbow trout linkage maps was found for both gene syntenies and orders. CONCLUSION: This new map is likely to cover the whole set of chromosome arms and should provide a useful framework to integrate existing or forthcoming rainbow trout linkage maps and other genomic resources. Since very large numbers of EST containing microsatellite sequences are available in databases, it becomes feasible to construct high-density linkage maps localizing known genes. This will facilitate comparative mapping and, eventually, identification of candidate genes in QTL studies. PMID- 17137493 TI - Dietary carbohydrate source influences molecular fingerprints of the rat faecal microbiota. AB - BACKGROUND: A study was designed to elucidate effects of selected carbohydrates on composition and activity of the intestinal microbiota. Five groups of eight rats were fed a western type diet containing cornstarch (reference group), sucrose, potato starch, inulin (a long- chained fructan) or oligofructose (a short-chained fructan). Fructans are, opposite sucrose and starches, not digestible by mammalian gut enzymes, but are known to be fermentable by specific bacteria in the large intestine. RESULTS: Animals fed with diets containing potato starch, or either of the fructans had a significantly (p < 0.05) higher caecal weight and lower caecal pH when compared to the reference group, indicating increased fermentation. Selective cultivation from faeces revealed a higher amount of lactic acid bacteria cultivable on Rogosa agar in these animals. Additionally, the fructan groups had a lower amount of coliform bacteria in faeces. In the inulin and oligofructose groups, higher levels of butyrate and propionate, respectively, were measured.Principal Component Analysis of profiles of the faecal microbiota obtained by Denaturing Gradient Gel Electrophoresis (DGGE) of PCR amplified bacterial 16S rRNA genes as well as of Reverse Transcriptase-PCR amplified bacterial 16S rRNA resulted in different phylogenetic profiles for each of the five animal groups as revealed by Principal Component Analysis (PCA) of band patterns. CONCLUSION: Even though sucrose and cornstarch are both easily digestible and are not expected to reach the large intestine, the DGGE band patterns obtained indicated that these carbohydrates indeed affected the composition of bacteria in the large gut. Also the two fructans resulted in completely different molecular fingerprints of the faecal microbiota, indicating that even though they are chemically similar, different intestinal bacteria ferment them. Comparison of DNA-based and RNA-based profiles suggested that two species within the phylum Bacteroidetes were not abundant in numbers but had a particularly high ribosome content in the animals fed with inulin. PMID- 17137494 TI - Levels and trends of demographic indices in southern rural Mozambique: evidence from demographic surveillance in Manhica district. AB - BACKGROUND: In Mozambique most of demographic data are obtained using census or sample survey including indirect estimations. A method of collecting longitudinal demographic data was introduced in southern Mozambique since 1996 (DSS Demographic Surveillance System in Manhica district, Maputo province), but the extent to which it yields demographic measures that are typical of southern rural Mozambique has not been evaluated yet. METHODS: Data from the DSS were used to estimate the levels and trends of fertility, mortality and migration in Manhica, between 1998 and 2005. The estimates from Manhica were compared with estimates from Maputo province using the 1997 National census and 1997 Demographic and Health Survey (DHS). The DHS data were used to estimate levels and trends of adult mortality using the siblings' histories and the orphanhood methods. RESULTS: The populations in Manhica and in Maputo province are young (44% <15 years in Manhica and 42% in Maputo); with reduced adult males when compared to females (all ages sex ratio of 78.7 in Manhica and 89 in Maputo). Fertility in Manhica is at a similar level as in Maputo province and has remained around 5 children per woman, during the eight years of surveillance in Manhica. Although the infant mortality rate (IMR) in Mozambique has decreased during the last two decades (from 148 deaths per 1000 live births in 1980 to 101 in 2003), it has remained stable around 80 in Manhica during the surveillance period. Adult mortality has increased both in Manhica (probability of dying from ages 15 to 60 increased from 0.4 in 1998 to 0.6 in 2005 in Manhica, from 0.3 in 1992 to 0.4 in 1997 in Maputo province and from 0.1 in 1980 to 0.6 in 2000 in Mozambique). Consequently, the life expectancy decreased from 53 to 46 in Manhica and from 42 years in 1997 to 38 in 2004 in Mozambique. Migration is high in Manhica but tends to stabilise after the movements of resettlement that followed the end of the civil war in 1992. CONCLUSION: The population under demographic surveillance in Manhica district presents characteristics that are typical of southern rural Mozambique, with predominance of young people and reduction of adult males. Labour migration and excess adult male mortality are the major factors for the reduction of adult males. Mortality is high and only infant mortality has started to stabilise while adult mortality has increased, and as consequence, life expectancy has decreased. The Manhica DSS is an adequate tool to report demographic measures for southern rural Mozambique. PMID- 17137495 TI - Study protocol to investigate the effects of testosterone therapy as an adjunct to exercise rehabilitation in hypogonadal males with chronic heart failure. AB - BACKGROUND: Testosterone deficiency is a common occurrence in men with chronic heart failure (CHF) and may underpin features of advanced disease, including reduced skeletal muscle mass and fatigue. It is positively correlated with cardiac output and exercise capacity in patients with CHF, whereas a significant improvement in both these parameters has been observed following testosterone replacement therapy. Testosterone therapy has also been shown to reduce circulating levels of inflammatory markers, (TNF-alpha, sICAM-1 and sVCAM-1) in patients with established coronary artery disease and testosterone deficiency. This pilot study will assess the feasibility of a combined exercise rehabilitation and adjunctive testosterone therapy intervention for evoking improvements in exercise capacity, circulating inflammatory markers, cardiac and skeletal muscle function, indices of psychological health status and quality of life in hypogonadal males with chronic heart failure. METHODS/DESIGN: Following ethical approval, 36 patients will be randomly allocated to one of two groups: testosterone or placebo therapy during exercise rehabilitation. A combined programme of moderate intensity aerobic exercise and resistance (strength) training will be used. The primary outcome measure is exercise capacity, assessed using an incremental shuttle walk test. Secondary outcome measures include measures of peak oxygen uptake, cardiac function, lower-limb skeletal muscle contractile function and oxygenation during exercise, circulating inflammatory markers, psychological health status and quality of life. DISCUSSION: Exercise rehabilitation can safely increase exercise capacity in stable CHF patients but there is a need for studies which are aimed at evaluating the long-term effects of physical training on functional status, morbidity and mortality. This pilot study will provide valuable preliminary data on the efficacy of testosterone therapy as an adjunct to exercise rehabilitation on a range of functional, physiological and health-related outcomes in this patient population. Preliminary data will be used in the design of a large-scale randomised controlled trial, aimed at informing clinical practice with respect to optimisation of exercise rehabilitation in this patient group. PMID- 17137496 TI - Synthesis of coumarin or ferrocene labeled nucleosides via Staudinger ligation. AB - BACKGROUND: Reaction of azides with triaryl phosphines under mild conditions gives iminophosphoranes which can react with almost any kind of electrophilic reagent, e.g. aldehydes/ketones to form imines or esters to form amides. This so called Staudinger ligation has been employed in a wide range of applications as a general tool for bioconjugation including specific labeling of nucleic acids. RESULTS: A new approach for the preparation of labeled nucleosides via intermolecular Staudinger ligation is described. Reaction of azidonucleosides with triphenylphosphine lead to iminophosphorane intermediates, which react subsequently with derivatives of coumarin or ferrocene to form coumarin or ferrocene labeled nucleosides. Fluorescent properties of coumarin labeled nucleosides are determined. CONCLUSION: New coumarin and ferrocene labeled nucleosides were prepared via intermolecular Staudinger ligation. This reaction joins the fluorescent coumarin and biospecific nucleoside to the new molecule with promising fluorescent and electrochemical properties. The isolated yields of products depend on the structure of azidonucleoside and carboxylic acids. A detailed study of the kinetics of the Staudinger ligation with nucleoside substrates is in progress. PMID- 17137497 TI - An accurate and interpretable model for siRNA efficacy prediction. AB - BACKGROUND: The use of exogenous small interfering RNAs (siRNAs) for gene silencing has quickly become a widespread molecular tool providing a powerful means for gene functional study and new drug target identification. Although considerable progress has been made recently in understanding how the RNAi pathway mediates gene silencing, the design of potent siRNAs remains challenging. RESULTS: We propose a simple linear model combining basic features of siRNA sequences for siRNA efficacy prediction. Trained and tested on a large dataset of siRNA sequences made recently available, it performs as well as more complex state-of-the-art models in terms of potency prediction accuracy, with the advantage of being directly interpretable. The analysis of this linear model allows us to detect and quantify the effect of nucleotide preferences at particular positions, including previously known and new observations. We also detect and quantify a strong propensity of potent siRNAs to contain short asymmetric motifs in their sequence, and show that, surprisingly, these motifs alone contain at least as much relevant information for potency prediction as the nucleotide preferences for particular positions. CONCLUSION: The model proposed for prediction of siRNA potency is as accurate as a state-of-the-art nonlinear model and is easily interpretable in terms of biological features. It is freely available on the web at http://cbio.ensmp.fr/dsir. PMID- 17137498 TI - Inhibition of inducible Nitric Oxide Synthase by a mustard gas analog in murine macrophages. AB - BACKGROUND: 2-Chloroethyl ethyl sulphide (CEES) is a sulphur vesicating agent and an analogue of the chemical warfare agent 2,2'-dichlorodiethyl sulphide, or sulphur mustard gas (HD). Both CEES and HD are alkylating agents that influence cellular thiols and are highly toxic. In a previous publication, we reported that lipopolysaccharide (LPS) enhances the cytotoxicity of CEES in murine RAW264.7 macrophages. In the present investigation, we studied the influence of CEES on nitric oxide (NO) production in LPS stimulated RAW264.7 cells since NO signalling affects inflammation, cell death, and wound healing. Murine macrophages stimulated with LPS produce NO almost exclusively via inducible nitric oxide synthase (iNOS) activity. We suggest that the influence of CEES or HD on the cellular production of NO could play an important role in the pathophysiological responses of tissues to these toxicants. In particular, it is known that macrophage generated NO synthesised by iNOS plays a critical role in wound healing. RESULTS: We initially confirmed that in LPS stimulated RAW264.7 macrophages NO is exclusively generated by the iNOS form of nitric oxide synthase. CEES treatment inhibited the synthesis of NO (after 24 hours) in viable LPS-stimulated RAW264.7 macrophages as measured by either nitrite secretion into the culture medium or the intracellular conversion of 4,5-diaminofluorescein diacetate (DAF-2DA) or dichlorofluorescin diacetate (DCFH-DA). Western blots showed that CEES transiently decreased the expression of iNOS protein; however, treatment of active iNOS with CEES in vitro did not inhibit its enzymatic activity CONCLUSION: CEES inhibits NO production in LPS stimulated macrophages by decreasing iNOS protein expression. Decreased iNOS expression is likely the result of CEES induced alteration in the nuclear factor kappa B (NF-kappaB) signalling pathway. Since NO can act as an antioxidant, the CEES induced down regulation of iNOS in LPS-stimulated macrophages could elevate oxidative stress. Since macrophage generated NO is known to play a key role in cutaneous wound healing, it is possible that this work has physiological relevance with respect to the healing of HD induced skin blisters. PMID- 17137499 TI - Cerebral edema in a patient following cytoreductive surgery and hyperthermic intraoperative intraperitoneal chemoperfusion. AB - BACKGROUND: Cytoreductive surgery and intraoperative, intraperitoneal hyperthermic chemoperfusion (HIPEC) is increasingly used to treat peritoneal surface metastases. We describe a fatal case of cerebral edema in a patient with appendiceal carcinoma and an underlying seizure disorder who underwent cytoreductive surgery and HIPEC. CASE PRESENTATION: A case of fatal postoperative cerebral edema is presented in a patient with an underlying seizure disorder and recurrent mucinous adenocarcinoma of the appendix. The patient was treated with cytoreductive surgery and intraoperative intraperitoneal hyperthermic chemoperfusion. The details and implications of this complication are discussed. CONCLUSION: The recognition of this potential complication is important for physicians performing cytoreductive surgery and HIPEC. Special caution should be taken when patients with seizure disorders are being considered for this treatment. PMID- 17137500 TI - Progression of pulmonary hyperinflation and trapped gas associated with genetic and environmental factors in children with cystic fibrosis. AB - BACKGROUND: Functional deterioration in cystic fibrosis (CF) may be reflected by increasing bronchial obstruction and, as recently shown, by ventilation inhomogeneities. This study investigated which physiological factors (airway obstruction, ventilation inhomogeneities, pulmonary hyperinflation, development of trapped gas) best express the decline in lung function, and what role specific CFTR genotypes and different types of bronchial infection may have upon this process. METHODS: Serial annual lung function tests, performed in 152 children (77 males; 75 females) with CF (age range: 6-18 y) provided data pertaining to functional residual capacity (FRCpleth, FRCMBNW), volume of trapped gas (VTG), effective specific airway resistance (sReff), lung clearance index (LCI), and forced expiratory indices (FVC, FEV1, FEF50). RESULTS: All lung function parameters showed progression with age. Pulmonary hyperinflation (FRCpleth > 2SDS) was already present in 39% of patients at age 6-8 yrs, increasing to 67% at age 18 yrs. The proportion of patients with VTG > 2SDS increased from 15% to 54% during this period. Children with severe pulmonary hyperinflation and trapped gas at age 6-8 yrs showed the most pronounced disease progression over time. Age related tracking of lung function parameters commences early in life, and is significantly influenced by specific CFTR genotypes. The group with chronic P. aeruginosa infection demonstrated most rapid progression in all lung function parameters, whilst those with chronic S. aureus infection had the slowest rate of progression. LCI, measured as an index of ventilation inhomogeneities was the most sensitive discriminator between the 3 types of infection examined (p < 0.0001). CONCLUSION: The relationships between lung function indices, CFTR genotypes and infective organisms observed in this study suggest that measurement of other lung function parameters, in addition to spirometry alone, may provide important information about disease progression in CF. PMID- 17137501 TI - Maternal position during the first stage of labor: a systematic review. AB - BACKGROUND: Policy makers and health professionals are progressively using evidence-based rationale to guide their decisions. There has long been controversy regarding which maternal position is more appropriate during the first stage of labor. This problem has been examined often and repeatedly and the optimal recommendation remains unclear. METHODS: This is a systematic review of the effect of maternal position during the first stage of labor. The main question addressed here is: Does encouraging women to adopt an upright position or to ambulate during the first stage of labor reduce the duration of this stage? All randomized controlled trials carried out to assess this effect were taken into consideration in this review. The following electronic databases were accessed to identify studies: MEDLINE, Popline, the Scientific Electronic Library On-line and the Latin American and Caribbean Health Science Information. Citation eligibility was independently assessed by two reviewers. The methodological quality of each trial was also evaluated independently by two reviewers and a trial under consideration was included only when consensus had been attained. Allocation concealment and screening for the occurrence of attrition, performance and detection biases were considered when studies were appraised. The decision whether to perform data pooling was based on the clinical similarity of studies. RESULTS: The search strategy resulted in 260 citations, of which 18 were assessed in full-text. Nine eligible randomized controlled trials were included in the systematic review. Randomization methods were not fully described in eight studies. The allocation concealment was considered adequate in four studies and unclear in five. The investigators pooled the data from seven studies in which the length of the first stage of labor and results were in favor of the intervention, but the high level of heterogeneity (I2 = 88.4%) impaired the meaning of this finding. The intervention did not affect other outcomes studied (mode of delivery, use of analgesia, labor augmentation and condition of the child at birth). CONCLUSION: Adoption of the upright position or ambulation during first stage of labor may be safe, but considering the available evidence and its consistency, it cannot be recommended as an effective intervention to reduce duration of the first stage of labor. PMID- 17137502 TI - Dynamic variable selection in SNP genotype autocalling from APEX microarray data. AB - BACKGROUND: Single nucleotide polymorphisms (SNPs) are DNA sequence variations, occurring when a single nucleotide--adenine (A), thymine (T), cytosine (C) or guanine (G)--is altered. Arguably, SNPs account for more than 90% of human genetic variation. Our laboratory has developed a highly redundant SNP genotyping assay consisting of multiple probes with signals from multiple channels for a single SNP, based on arrayed primer extension (APEX). This mini-sequencing method is a powerful combination of a highly parallel microarray with distinctive Sanger based dideoxy terminator sequencing chemistry. Using this microarray platform, our current genotype calling system (known as SNP Chart) is capable of calling single SNP genotypes by manual inspection of the APEX data, which is time consuming and exposed to user subjectivity bias. RESULTS: Using a set of 32 Coriell DNA samples plus three negative PCR controls as a training data set, we have developed a fully-automated genotyping algorithm based on simple linear discriminant analysis (LDA) using dynamic variable selection. The algorithm combines separate analyses based on the multiple probe sets to give a final posterior probability for each candidate genotype. We have tested our algorithm on a completely independent data set of 270 DNA samples, with validated genotypes, from patients admitted to the intensive care unit (ICU) of St. Paul's Hospital (plus one negative PCR control sample). Our method achieves a concordance rate of 98.9% with a 99.6% call rate for a set of 96 SNPs. By adjusting the threshold value for the final posterior probability of the called genotype, the call rate reduces to 94.9% with a higher concordance rate of 99.6%. We also reversed the two independent data sets in their training and testing roles, achieving a concordance rate up to 99.8%. CONCLUSION: The strength of this APEX chemistry-based platform is its unique redundancy having multiple probes for a single SNP. Our model-based genotype calling algorithm captures the redundancy in the system considering all the underlying probe features of a particular SNP, automatically down-weighting any 'bad data' corresponding to image artifacts on the microarray slide or failure of a specific chemistry. In this regard, our method is able to automatically select the probes which work well and reduce the effect of other so-called bad performing probes in a sample-specific manner, for any number of SNPs. PMID- 17137503 TI - Gelatinases, endonuclease and Vascular Endothelial Growth Factor during development and regression of swine luteal tissue. AB - BACKGROUND: The development and regression of corpus luteum (CL) is characterized by an intense angiogenesis and angioregression accompanied by luteal tissue and extracellular matrix (ECM) remodelling. Vascular Endothelial Growth Factor (VEGF) is the main regulator of angiogenesis, promoting endothelial cell mitosis and differentiation. After the formation of neovascular tubes, the remodelling of ECM is essential for the correct development of CL, particularly by the action of specific class of proteolytic enzymes known as matrix metalloproteinases (MMPs). During luteal regression, characterized by an apoptotic process and successively by an intense ECM and luteal degradation, the activation of Ca++/Mg++-dependent endonucleases and MMPs activity are required. The levels of expression and activity of VEGF, MMP-2 and -9, and Ca++/Mg++-dependent endonucleases throughout the oestrous cycle and at pregnancy were analyzed. RESULTS: Different patterns of VEGF, MMPs and Ca++/Mg++-dependent endonuclease were observed in swine CL during different luteal phases and at pregnancy. Immediately after ovulation, the highest levels of VEGF mRNA/protein and MMP-9 activity were detected. On days 5 14 after ovulation, VEGF expression and MMP-2 and -9 activities are at basal levels, while Ca++/Mg++-dependent endonuclease levels increased significantly in relation to day 1. Only at luteolysis (day 17), Ca++/Mg++-dependent endonuclease and MMP-2 spontaneous activity increased significantly. At pregnancy, high levels of MMP-9 and VEGF were observed. CONCLUSION: Our findings, obtained from a precisely controlled in vivo model of CL development and regression, allow us to determine relationships among VEGF, MMPs and endonucleases during angiogenesis and angioregression. Thus, CL provides a very interesting model for studying factors involved in vascular remodelling. PMID- 17137504 TI - Geostatistical analysis of disease data: accounting for spatial support and population density in the isopleth mapping of cancer mortality risk using area-to point Poisson kriging. AB - BACKGROUND: Geostatistical techniques that account for spatially varying population sizes and spatial patterns in the filtering of choropleth maps of cancer mortality were recently developed. Their implementation was facilitated by the initial assumption that all geographical units are the same size and shape, which allowed the use of geographic centroids in semivariogram estimation and kriging. Another implicit assumption was that the population at risk is uniformly distributed within each unit. This paper presents a generalization of Poisson kriging whereby the size and shape of administrative units, as well as the population density, is incorporated into the filtering of noisy mortality rates and the creation of isopleth risk maps. An innovative procedure to infer the point-support semivariogram of the risk from aggregated rates (i.e. areal data) is also proposed. RESULTS: The novel methodology is applied to age-adjusted lung and cervix cancer mortality rates recorded for white females in two contrasted county geographies: 1) state of Indiana that consists of 92 counties of fairly similar size and shape, and 2) four states in the Western US (Arizona, California, Nevada and Utah) forming a set of 118 counties that are vastly different geographical units. Area-to-point (ATP) Poisson kriging produces risk surfaces that are less smooth than the maps created by a naive point kriging of empirical Bayesian smoothed rates. The coherence constraint of ATP kriging also ensures that the population-weighted average of risk estimates within each geographical unit equals the areal data for this unit. Simulation studies showed that the new approach yields more accurate predictions and confidence intervals than point kriging of areal data where all counties are simply collapsed into their respective polygon centroids. Its benefit over point kriging increases as the county geography becomes more heterogeneous. CONCLUSION: A major limitation of choropleth maps is the common biased visual perception that larger rural and sparsely populated areas are of greater importance. The approach presented in this paper allows the continuous mapping of mortality risk, while accounting locally for population density and areal data through the coherence constraint. This form of Poisson kriging will facilitate the analysis of relationships between health data and putative covariates that are typically measured over different spatial supports. PMID- 17137505 TI - INSIG-2 promoter polymorphism and obesity related phenotypes: association study in 1428 members of 248 families. AB - BACKGROUND: Obesity is a major public health problem. Body mass index (BMI) is a highly heritable phenotype but robust associations of genetic polymorphisms to BMI or other obesity-related phenotypes have been difficult to establish. Recently a large genetic association study showed evidence for association of the single nucleotide polymorphism (SNP) rs7566605, which lies 10 Kb 5' to the first exon of the insulin-induced gene 2 (INSIG-2), with obesity in several cohorts. We tested this polymorphism for association with body mass related phenotypes in a large family study whose mean BMI was consistent with moderate overweight. METHODS: We studied 1428 members of 248 British Caucasian families who had been ascertained through a proband with hypertension. We measured BMI, waist and hip circumference, and plasma levels of leptin. We genotyped the rs7566605 SNP using a restriction fragment length polymorphism assay, and carried out a family-based association test for quantitative traits related to obesity using the statistical programs MERLIN and QTDT. RESULTS: We observed no significant association between genotype at rs7566605 and covariate-adjusted (for age, sex, alcohol consumption, smoking and exercise habit) log-transformed BMI, waist measurement, hip measurement, waist-to-hip ratio, or plasma levels of leptin. CONCLUSION: There was no association between genotype at rs7566605 and obesity-related phenotypes in this British Caucasian population. These families were in general moderately overweight, few members being severely obese. Our result indicates that this polymorphism has little if any effect on BMI within the normal to moderately overweight range. The effects of this polymorphism on body mass may be restricted to those already predisposed to at least moderate obesity as a result of environmental factors and other predisposing genotypes. PMID- 17137507 TI - LRRK2 in Parkinson's disease and dementia with Lewy bodies. AB - BACKGROUND: Mutations in LRRK2 encoding leucine-rich repeat kinase 2 are thus far the most frequent genetic cause associated with autosomal dominant and idiopathic Parkinson's disease (PD). To examine whether LRRK2 is directly associated with neuropathology of PD and other related disorders, we analyzed LRRK2 in brains of patients affected by PD and dementia with Lewy bodies (DLB) using highly specific antibodies to LRRK2. RESULTS: We demonstrated that anti-LRRK2 antibodies strongly labelled brainstem and cortical Lewy bodies, the pathological hallmarks of PD and DLB, respectively. In addition, anti-LRRK2 also labelled brain vasculature, axons, and neuronal cell bodies. Interestingly, the immunocytochemical profile of LRRK2 varied with different antibodies depending upon specific antigenic sites along the LRRK2 protein. All anti-LRRK2 antibodies tested that were raised against various regions of LRRK2, were found to be immunoreactive to recombinant LRRK2 on Western blots. However, only the antibodies raised against the N terminal and C-terminal regions of LRRK2, but not the regions containing folded protein domains, were positive in immunolabeling of Lewy bodies, suggesting a differential exposure of specific antigenic sites of LRRK2 on tissue sections. CONCLUSION: We conclude that LRRK2 is a component of Lewy bodies in both PD and DLB, and therefore plays an important role in the Lewy body formation and disease pathogenesis. Information on the cellular localization of LRRK2 under normal and pathological conditions will deepen our understanding of its functions and molecular pathways relevant to the progression of PD and related disorders. PMID- 17137506 TI - FGFR1 and WT1 are markers of human prostate cancer progression. AB - BACKGROUND: Androgen-independent prostate adenocarcinomas are responsible for about 6% of overall cancer deaths in men. METHODS: We used DNA microarrays to identify genes related to the transition between androgen-dependent and androgen independent stages in the LuCaP 23.1 xenograft model of prostate adenocarcinoma. The expression of the proteins encoded by these genes was then assessed by immunohistochemistry on tissue microarrays (TMA) including human prostate carcinoma samples issued from 85 patients who had undergone radical prostatectomy. RESULTS: FGFR1, TACC1 and WT1 gene expression levels were associated with the androgen-independent stage in xenografts and human prostate carcinoma samples. MART1 protein expression was correlated with pT2 tumor stages. CONCLUSION: Our results suggest that each of these four genes may play a role, or at least reflect a stage of prostate carcinoma growth/development/progression. PMID- 17137508 TI - Genomewide identification of pheromone-targeted transcription in fission yeast. AB - BACKGROUND: Fission yeast cells undergo sexual differentiation in response to nitrogen starvation. In this process haploid M and P cells first mate to form diploid zygotes, which then enter meiosis and sporulate. Prior to mating, M and P cells communicate with diffusible mating pheromones that activate a signal transduction pathway in the opposite cell type. The pheromone signalling orchestrates mating and is also required for entry into meiosis. RESULTS: Here we use DNA microarrays to identify genes that are induced by M-factor in P cells and by P-factor in M-cells. The use of a cyr1 genetic background allowed us to study pheromone signalling independently of nitrogen starvation. We identified a total of 163 genes that were consistently induced more than two-fold by pheromone stimulation. Gene disruption experiments demonstrated the involvement of newly discovered pheromone-induced genes in the differentiation process. We have mapped Gene Ontology (GO) categories specifically associated with pheromone induction. A direct comparison of the M- and P-factor induced expression pattern allowed us to identify cell-type specific transcripts, including three new M-specific genes and one new P-specific gene. CONCLUSION: We found that the pheromone response was very similar in M and P cells. Surprisingly, pheromone control extended to genes fulfilling their function well beyond the point of entry into meiosis, including numerous genes required for meiotic recombination. Our results suggest that the Ste11 transcription factor is responsible for the majority of pheromone-induced transcription. Finally, most cell-type specific genes now appear to be identified in fission yeast. PMID- 17137509 TI - Cis-motifs upstream of the transcription and translation initiation sites are effectively revealed by their positional disequilibrium in eukaryote genomes using frequency distribution curves. AB - BACKGROUND: The discovery of cis-regulatory motifs still remains a challenging task even though the number of sequenced genomes is constantly growing. Computational analyses using pattern search algorithms have been valuable in phylogenetic footprinting approaches as have expression profile experiments to predict co-occurring motifs. Surprisingly little is known about the nature of cis regulatory element (CRE) distribution in promoters. RESULTS: In this paper we used the Motif Mapper open-source collection of visual basic scripts for the analysis of motifs in any aligned set of DNA sequences. We focused on promoter motif distribution curves to identify positional over-representation of DNA motifs. Using differentially aligned datasets from the model species Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster and Saccharomyces cerevisiae, we convincingly demonstrated the importance of the position and orientation for motif discovery. Analysis with known CREs and all possible hexanucleotides showed that some functional elements gather close to the transcription and translation initiation sites and that elements other than the TATA-box motif are conserved between eukaryote promoters. While a high background frequency usually decreases the effectiveness of such an enumerative investigation, we improved our analysis by conducting motif distribution maps using large datasets. CONCLUSION: This is the first study to reveal positional over-representation of CREs and promoter motifs in a cross-species approach. CREs and motifs shared between eukaryotic promoters support the observation that an eukaryotic promoter structure has been conserved throughout evolutionary time. Furthermore, with the information on positional enrichment of a motif or a known functional CRE, it is possible to get a more detailed insight into where an element appears to function. This in turn might accelerate the in depth examination of known and yet unknown cis-regulatory sequences in the laboratory. PMID- 17137510 TI - Genome-wide identification of functionally distinct subsets of cellular mRNAs associated with two nucleocytoplasmic-shuttling mammalian splicing factors. AB - BACKGROUND: Pre-mRNA splicing is an essential step in gene expression that occurs co-transcriptionally in the cell nucleus, involving a large number of RNA binding protein splicing factors, in addition to core spliceosome components. Several of these proteins are required for the recognition of intronic sequence elements, transiently associating with the primary transcript during splicing. Some protein splicing factors, such as the U2 small nuclear RNP auxiliary factor (U2AF), are known to be exported to the cytoplasm, despite being implicated solely in nuclear functions. This observation raises the question of whether U2AF associates with mature mRNA-ribonucleoprotein particles in transit to the cytoplasm, participating in additional cellular functions. RESULTS: Here we report the identification of RNAs immunoprecipitated by a monoclonal antibody specific for the U2AF 65 kDa subunit (U2AF65) and demonstrate its association with spliced mRNAs. For comparison, we analyzed mRNAs associated with the polypyrimidine tract binding protein (PTB), a splicing factor that also binds to intronic pyrimidine rich sequences but additionally participates in mRNA localization, stability, and translation. Our results show that 10% of cellular mRNAs expressed in HeLa cells associate differentially with U2AF65 and PTB. Among U2AF65-associated mRNAs there is a predominance of transcription factors and cell cycle regulators, whereas PTB associated transcripts are enriched in mRNA species that encode proteins implicated in intracellular transport, vesicle trafficking, and apoptosis. CONCLUSION: Our results show that U2AF65 associates with specific subsets of spliced mRNAs, strongly suggesting that it is involved in novel cellular functions in addition to splicing. PMID- 17137511 TI - The pulmonary effects of intravenous adenosine in asthmatic subjects. AB - BACKGROUND: We have shown that intravenous adenosine in normal subjects does not cause bronchospasm, but causes dyspnea, most likely by an effect on vagal C fibers in the lungs [Burki et al. J Appl Physiol 2005; 98:180-5]. Since airways inflammation and bronchial hyperreactivity are features of asthma, it is possible that intravenous adenosine may be associated with an increased intensity of dyspnea, and may cause bronchospasm, as noted anecdotally in previous reports. METHODS: We compared the effects of placebo and 10 mg intravenous adenosine, in 6 normal and 6 asthmatic subjects. RESULTS: Placebo injection had no significant (p > 0.05) effect on the forced expiratory spirogram, heart rate, minute ventilation (Ve), or respiratory sensation. Similarly, adenosine injection caused no significant changes (p > 0.05) in the forced expiratory spirogram; however, there was a rapid development of dyspnea as signified visually on a modified Borg scale, and a significant (p < 0.05) tachycardia in each subject (Asthmatics +18%, Normals + 34%), and a significant (p < 0.05) increase in Ve (Asthmatics +93%, Normals +130%). The intensity of dyspnea was significantly greater (p < 0.05) in the asthmatic subjects. CONCLUSION: These data indicate that intravenous adenosine does not cause bronchospasm in asthmatic subjects, and supports the concept that adenosine-induced dyspnea is most likely secondary to stimulation of vagal C fibers in the lungs. The increased intensity of adenosine-induced dyspnea in the asthmatic subjects suggests that airways inflammation may have sensitized the vagal C fibers. PMID- 17137512 TI - A population-based study examining the emergence of community-associated methicillin-resistant Staphylococcus aureus USA300 in New York City. AB - BACKGROUND: Community-associated methicillin-resistant Staphylococcus aureus (CA MRSA) is a serious pathogen in several regions in the United States. It is unclear which populations are at high risk for the emergence of these strains. METHODS: All unique patient isolates of S. aureus were collected from hospitals in Brooklyn, NY over a three-month period. Isolates of MRSA that were susceptible to clindamycin underwent SCCmec typing. Isolates with the SCCmec type IV (characteristic of CA-MRSA strains) underwent ribotyping. Demographic information involving the neighborhoods of Brooklyn was also gathered and correlated with the prevalence of CA-MRSA strains. RESULTS: Of 1316 isolates collected during the surveillance, 217 were MRSA susceptible to clindamycin. A total of 125 isolates possessed SCCmec type IV; 72 belonged to the USA300 strain and five belonged to the USA400 strain. Hospitals in the eastern part of the city had the highest prevalence of USA300 strain. Individuals in the eastern region, when compared to the western region, were more likely to be Black, Hispanic, female, and < 18 years of age, and to have households of > or = 3 persons. In addition, the median household income was lower, and the proportion of individuals on public assistance was higher, for the population in the eastern region. CONCLUSION: The USA300 strain of CA-MRSA is emerging in New York City. In this population-based study, urban regions of lower socioeconomic status and with evidence of overcrowding appear to be at higher risk for the emergence of this pathogen. PMID- 17137513 TI - Male circumcision, religion, and infectious diseases: an ecologic analysis of 118 developing countries. AB - BACKGROUND: Both religious practices and male circumcision (MC) have been associated with HIV and other sexually-transmitted infectious diseases. Most studies have been limited in size and have not adequately controlled for religion, so these relationships remain unclear. METHODS: We evaluated relationships between MC prevalence, Muslim and Christian religion, and 7 infectious diseases using country-specific data among 118 developing countries. We used multivariate linear regression to describe associations between MC and cervical cancer incidence, and between MC and HIV prevalence among countries with primarily sexual HIV transmission. RESULTS: Fifty-three, 14, and 51 developing countries had a high (>80%), intermediate (20-80%), and low (<20%) MC prevalence, respectively. In univariate analyses, MC was associated with lower HIV prevalence and lower cervical cancer incidence, but not with HSV-2, syphilis, nor, as expected, with Hepatitis C, tuberculosis, or malaria. In multivariate analysis after stratifying the countries by religious groups, each categorical increase of MC prevalence was associated with a 3.65/100,000 women (95% CI 0.54-6.76, p = 0.02) decrease in annual cervical cancer incidence, and a 1.84-fold (95% CI 1.36 2.48, p < 0.001) decrease in the adult HIV prevalence among sub-Saharan African countries. In separate multivariate analyses among non-sub-Saharan African countries controlling for religion, higher MC prevalence was associated with a 8.94-fold (95% CI 4.30-18.60) decrease in the adult HIV prevalence among countries with primarily heterosexual HIV transmission, but not, as expected, among countries with primarily homosexual or injection drug use HIV transmission (p = 0.35). CONCLUSION: Male circumcision was significantly associated with lower cervical cancer incidence and lower HIV prevalence in sub-Saharan Africa, independent of Muslim and Christian religion. As predicted, male circumcision was also strongly associated with lower HIV prevalence among countries with primarily heterosexual HIV transmission, but not among countries with primarily homosexual or injection drug use HIV transmission. These findings strengthen the reported biological link between MC and some sexually transmitted infectious diseases, including HIV and cervical cancer. PMID- 17137514 TI - Low-frequency vibratory exercise reduces the risk of bone fracture more than walking: a randomized controlled trial. AB - BACKGROUND: Whole-body vibration (WBV) is a new type of exercise that has been increasingly tested for the ability to prevent bone fractures and osteoporosis in frail people. There are two currently marketed vibrating plates: a) the whole plate oscillates up and down; b) reciprocating vertical displacements on the left and right side of a fulcrum, increasing the lateral accelerations. A few studies have shown recently the effectiveness of the up-and-down plate for increasing Bone Mineral Density (BMD) and balance; but the effectiveness of the reciprocating plate technique remains mainly unknown. The aim was to compare the effects of WBV using a reciprocating platform at frequencies lower than 20 Hz and a walking-based exercise programme on BMD and balance in post-menopausal women. METHODS: Twenty-eight physically untrained post-menopausal women were assigned at random to a WBV group or a Walking group. Both experimental programmes consisted of 3 sessions per week for 8 months. Each vibratory session included 6 bouts of 1 min (12.6 Hz in frequency and 3 cm in amplitude with 60 degrees of knee flexion) with 1 min rest between bouts. Each walking session was 55 minutes of walking and 5 minutes of stretching. Hip and lumbar BMD (g.cm-2) were measured using dual energy X-ray absorptiometry and balance was assessed by the blind flamingo test. ANOVA for repeated measurements was adjusted by baseline data, weight and age. RESULTS: After 8 months, BMD at the femoral neck in the WBV group was increased by 4.3% (P = 0.011) compared to the Walking group. In contrast, the BMD at the lumbar spine was unaltered in both groups. Balance was improved in the WBV group (29%) but not in the Walking group. CONCLUSION: The 8-month course of vibratory exercise using a reciprocating plate is feasible and is more effective than walking to improve two major determinants of bone fractures: hip BMD and balance. PMID- 17137515 TI - BioMoby extensions to the Taverna workflow management and enactment software. AB - BACKGROUND: As biology becomes an increasingly computational science, it is critical that we develop software tools that support not only bioinformaticians, but also bench biologists in their exploration of the vast and complex data-sets that continue to build from international genomic, proteomic, and systems-biology projects. The BioMoby interoperability system was created with the goal of facilitating the movement of data from one Web-based resource to another to fulfill the requirements of non-expert bioinformaticians. In parallel with the development of BioMoby, the European myGrid project was designing Taverna, a bioinformatics workflow design and enactment tool. Here we describe the marriage of these two projects in the form of a Taverna plug-in that provides access to many of BioMoby's features through the Taverna interface. RESULTS: The exposed BioMoby functionality aids in the design of "sensible" BioMoby workflows, aids in pipelining BioMoby and non-BioMoby-based resources, and ensures that end-users need only a minimal understanding of both BioMoby, and the Taverna interface itself. Users are guided through the construction of syntactically and semantically correct workflows through plug-in calls to the Moby Central registry. Moby Central provides a menu of only those BioMoby services capable of operating on the data-type(s) that exist at any given position in the workflow. Moreover, the plug-in automatically and correctly connects a selected service into the workflow such that users are not required to understand the nature of the inputs or outputs for any service, leaving them to focus on the biological meaning of the workflow they are constructing, rather than the technical details of how the services will interoperate. CONCLUSION: With the availability of the BioMoby plug-in to Taverna, we believe that BioMoby-based Web Services are now significantly more useful and accessible to bench scientists than are more traditional Web Services. PMID- 17137516 TI - Study of trunk asymmetry in normal children and adolescents. AB - The scoliometer readings in both standing and sitting position of 2071 children and adolescents (1099 boys and 972 girls) aged from 5 to 18 years old were studied. The angle of trunk rotation (ATR) was measured, in order to quantify the existing trunk asymmetry. Children and adolescents were divided in two groups according to the severity of trunk asymmetry. In the first group asymmetry was 1 to 6 degrees and in the second group was 7 or more degrees. Radiographic and leg length inequality evaluation were also performed in a number of children. The mean frequency of symmetric (ATR = 0 degrees) boys and girls was 67.06% and 65.01% for the standing screening position and 76.5% and 75.1% for the sitting position, respectively. The mean difference of frequency of asymmetry (ATR > 0 degrees) at standing minus sitting forward bending position for boys and girls was 10.22% and 9.37%, respectively. The mean frequency of asymmetry of 7 or more degrees was 3.23% for boys and 3.92% for girls at the standing forward bending position and 1.62% and 2.21% at the sitting, respectively. Girls are found to express higher frequency of asymmetry than boys. Right trunk asymmetry was more common than left. The sitting position is the preferred screening position for examining the rib or loin hump during school screening as it demonstrates the best correlation with the spinal deformity exposing the real trunk asymmetry. PMID- 17137517 TI - Key transmission parameters of an institutional outbreak during the 1918 influenza pandemic estimated by mathematical modelling. AB - AIM: To estimate the key transmission parameters associated with an outbreak of pandemic influenza in an institutional setting (New Zealand 1918). METHODS: Historical morbidity and mortality data were obtained from the report of the medical officer for a large military camp. A susceptible-exposed-infectious recovered epidemiological model was solved numerically to find a range of best fit estimates for key epidemic parameters and an incidence curve. Mortality data were subsequently modelled by performing a convolution of incidence distribution with a best-fit incidence-mortality lag distribution. RESULTS: Basic reproduction number (R0) values for three possible scenarios ranged between 1.3, and 3.1, and corresponding average latent period and infectious period estimates ranged between 0.7 and 1.3 days, and 0.2 and 0.3 days respectively. The mean and median best-estimate incidence-mortality lag periods were 6.9 and 6.6 days respectively. This delay is consistent with secondary bacterial pneumonia being a relatively important cause of death in this predominantly young male population. CONCLUSION: These R0 estimates are broadly consistent with others made for the 1918 influenza pandemic and are not particularly large relative to some other infectious diseases. This finding suggests that if a novel influenza strain of similar virulence emerged then it could potentially be controlled through the prompt use of major public health measures. PMID- 17137518 TI - Airway inflammation contributes to health status in COPD: a cross-sectional study. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is characterized by irreversible airflow limitation and airway inflammation, accompanied by decreased health status. It is still unknown which factors are responsible for the impaired health status in COPD. We postulated that airway inflammation negatively contributes to health status in COPD. METHODS: In 114 COPD patients (99 male, age: 62 +/- 8 yr, 41 [31-55] pack-years, no inhaled or oral corticosteroids, postbronchodilator FEV1: 63 +/- 9% pred, FEV1/IVC: 48 +/- 9%) we obtained induced sputum and measured health status (St. George's respiratory questionnaire (SGRQ)), postbronchodilator FEV1, hyperinflation (RV/TLC), and airway hyperresponsiveness to methacholine (PC20). Sputum was induced by hypertonic saline and differential cell counts were obtained in 102 patients. RESULTS: Univariate analysis showed that SGRQ total and symptom score were positively associated with % sputum macrophages (r = 0.20, p = 0.05; and r = 0.20, p = 0.04, respectively). Multiple regression analysis confirmed these relationships, providing significant contributions of % sputum macrophages (B = 0.25, p = 0.021) and RV/TLC (B = 0.60, p = 0.002) to SGRQ total score. Furthermore, SGRQ symptom score was associated with % sputum macrophages (B = 0.30, p = 0.03) and RV/TLC (B = 0.48, p = 0.044), whilst SGRQ activity score was associated with % sputum macrophages (B = 0.46, p = 0.002), RV/TLC (B = 0.61, p = 0.015), and PC20 (B = 9.3, p = 0.024). Current smoking and FEV1 were not significantly associated with health status in the multiple regression analysis. CONCLUSION: We conclude that worse health status in COPD patients is associated with higher inflammatory cell counts in induced sputum. Our findings suggest that airway inflammation and hyperinflation independently contribute to impaired health status in COPD. This may provide a rationale for anti-inflammatory therapy in this disease. PMID- 17137519 TI - Improvement in accuracy of multiple sequence alignment using novel group-to-group sequence alignment algorithm with piecewise linear gap cost. AB - BACKGROUND: Multiple sequence alignment (MSA) is a useful tool in bioinformatics. Although many MSA algorithms have been developed, there is still room for improvement in accuracy and speed. In the alignment of a family of protein sequences, global MSA algorithms perform better than local ones in many cases, while local ones perform better than global ones when some sequences have long insertions or deletions (indels) relative to others. Many recent leading MSA algorithms have incorporated pairwise alignment information obtained from a mixture of sources into their scoring system to improve accuracy of alignment containing long indels. RESULTS: We propose a novel group-to-group sequence alignment algorithm that uses a piecewise linear gap cost. We developed a program called PRIME, which employs our proposed algorithm to optimize the well-defined sum-of-pairs score. PRIME stands for Profile-based Randomized Iteration MEthod. We evaluated PRIME and some recent MSA programs using BAliBASE version 3.0 and PREFAB version 4.0 benchmarks. The results of benchmark tests showed that PRIME can construct accurate alignments comparable to the most accurate programs currently available, including L-INS-i of MAFFT, ProbCons, and T-Coffee. CONCLUSION: PRIME enables users to construct accurate alignments without having to employ pairwise alignment information. PRIME is available at http://prime.cbrc.jp/. PMID- 17137520 TI - Berberine chloride can ameliorate the spatial memory impairment and increase the expression of interleukin-1beta and inducible nitric oxide synthase in the rat model of Alzheimer's disease. AB - BACKGROUND: Berberine is the major alkaloidal component of Rhizoma coptidis, and has multiple pharmacological effects including inhibiting acetylcholinesterase, reducing cholesterol and glucose, lowering mortality in patients with chronic congestive heart failure and anti-inflammation etc. Thus berberine is a promising drug for diabetes, hyperlipemia, coronary artery disease and ischemic stroke etc. The present study was carried out to investigate the effect of berberine chloride on the spatial memory, inflammation factors interleukin-1 beta (IL-1beta) and inducible nitric oxide synthase (iNOS) expression in the rat model of Alzheimer's disease (AD) which was established by injecting Abeta (1-40) (5 microgram) into the rats hippocampuses bilaterally. RESULTS: The rats were given berberine chloride (50 mg/kg) by intragastric administration once daily for 14 days. The spatial memory was assayed by Morris water maze test, IL-1beta and iNOS in the hippocampus were assayed by immunohistochemistry and real time polymerase chain reaction (PCR). Intragastric administration of berberine significantly ameliorated the spatial memory impairment and increased the expression of IL 1beta, iNOS in the rat model of AD. CONCLUSION: Berberine might be beneficial to AD by intragastric administration though it might exaggerate the inflammation reaction. PMID- 17137521 TI - FV peptide induces apoptosis in HEp 2 and HeLa cells: an insight into the mechanism of induction. AB - The present study is an attempt to evaluate the antiproliferative potential of peptide (7.6 kDa) from lionfish (Pterios volitans) venom on cultured HEp2 and HeLa cells. Different dose of purified peptide (1, 2 and 4 microg/ml) at different time points (12, 24 and 36 hrs) were tested for antiproliferative index of the peptide. Among them, 2 microg/ml at 24 hrs was found to effectively inhibit cancer cell growth in vitro and did not cause any adverse effect on normal human lymphocytes. Apoptosis was examined by propidium iodide staining, confirmed by the expression of caspase-8 and caspase-3, down regulation of Bcl-2 expression and DNA fragmentation in treated cells, when compared to untreated HEp2 and HeLa cells. Thus fish venom peptide was found to selectively induce apoptosis in cancer cell. PMID- 17137522 TI - iHAP--integrated haplotype analysis pipeline for characterizing the haplotype structure of genes. AB - BACKGROUND: The advent of genotype data from large-scale efforts that catalog the genetic variants of different populations have given rise to new avenues for multifactorial disease association studies. Recent work shows that genotype data from the International HapMap Project have a high degree of transferability to the wider population. This implies that the design of genotyping studies on local populations may be facilitated through inferences drawn from information contained in HapMap populations. RESULTS: To facilitate analysis of HapMap data for characterizing the haplotype structure of genes or any chromosomal regions, we have developed an integrated web-based resource, iHAP. In addition to incorporating genotype and haplotype data from the International HapMap Project and gene information from the UCSC Genome Browser Database, iHAP also provides capabilities for inferring haplotype blocks and selecting tag SNPs that are representative of haplotype patterns. These include block partitioning algorithms, block definitions, tag SNP definitions, as well as SNPs to be "force included" as tags. Based on the parameters defined at the input stage, iHAP performs on-the-fly analysis and displays the result graphically as a webpage. To facilitate analysis, intermediate and final result files can be downloaded. CONCLUSION: The iHAP resource, available at http://ihap.bii.a-star.edu.sg, provides a convenient yet flexible approach for the user community to analyze HapMap data and identify candidate targets for genotyping studies. PMID- 17137523 TI - Detection of THCA in oral fluid by GC-MS-MS. AB - A major marijuana metabolite, 11-nor-9-carboxy-tetrahydrocannabinol (THCA), has been identified in oral fluids from donors that previously tested positive for Delta(9)-tetrahydrocannabinol (THC). The method consisted of solid-phase extraction of the oral fluid samples followed by gas chromatography-tandem mass spectrometry analysis of the extracts. Testing for THCA was performed on 223 oral fluid samples previously analyzed for THC. The THCA assay was linear from 10 to 240 pg/mL. The mean recovery of spiked THCA in oral fluid was 104%, and precision was 4% at 20 pg/mL using fortified negative samples. This method was rugged and robust, providing detection and quantification of THCA in oral fluids at levels not previously reported. Results of this study showed that THCA was detectable in 21 of 26 oral fluid samples previously reported positive for THC. The range of concentrations from these samples was from 10 pg/mL up to 142 pg/mL THCA. PMID- 17137524 TI - The influence of collection site and methods on postmortem morphine concentrations in a porcine model. AB - This study was to determine the relationship of antemortem to postmortem morphine concentrations in heart and femoral blood in a porcine model following acute intravenous opiate overdose. The study involved 20 swine; each was sacrificed 10 min after injection of 2 mg/kg body weight of morphine. Drug concentrations were assayed from vitreous humor and blood isolated from the femoral vein and artery and left and right ventricles at various times postmortem. Comparisons were made between antemortem and postmortem values to determine agreement and reliability. Both free and total postmortem values varied significantly among animals, sampling sites, and over time. Free postmortem values were generally higher in comparison with antemortem values, whereas postmortem total morphine values were similar to or slightly lower than antemortem values. The effect of time on postmortem values was small. These results demonstrate a significant amount of variability in free and total morphine measurements both over time and within and between sites. Furthermore, a comparison of antemortem to postmortem values demonstrates a lack of consistency relative to the dose of morphine administered. Concentrations of morphine in the femoral vein were typically the lowest observed. This observation is not surprising given the transformation that occurs prior to the drug reaching the femoral vein. Values associated with diffuse tissues, relative to femoral veins, demonstrate more stochastic variation. PMID- 17137525 TI - The effect of the use of mouthwash on ethylglucuronide concentrations in urine. AB - Two studies were performed to evaluate the effect of alcohol containing mouthwash on the appearance of ethyl glucuronide (EtG) in urine. In the first study, 9 volunteers were given a 4-oz bottle of mouthwash, which contained 12% ethanol. They gargled with all 4 oz. of the mouthwash at intervals over a 15-min period. All urine samples were collected over the next 24 h. Of 39 provided urine samples, there were 20 > 50 ng/mL, 12 > 100 ng/mL, 5 > 200 ng/mL, 3 > 250 ng/mL, and 1 > 300 ng/mL. The peak concentrations were all within 12 h after the exposure. In the second study, 11 participants gargled 3 times daily for 5 days. The first morning void was collected. Sixteen of the 55 submitted samples contained EtG concentrations of greater than 50 ng/mL. All of them were less than 120 ng/mL. These studies show that incidental exposure to mouthwash containing 12% ethanol, when gargling according to the manufacturer's instructions, can result in urinary EtG values greater than 50 ng/mL. All specimens were negative for ethanol. The limits of detection and quantitation for the EtG testing were 50 ng/mL. PMID- 17137526 TI - Detection of the administration of human erythropoietin (HuEPO) to canines. AB - Recombinant human erythropoietin (rHuEPO) doping is prohibited in animal (canine and equine) sport. The effectiveness of a range of immunoassay screening methods for the detection of rHuEPO in canine urine was evaluated. The excretion profiles following rHuEPO administration to dogs were investigated. The presence of rHuEPO in postadministration samples was confirmed using the World Anti-Doping Agency (WADA)-approved isoelectric focusing immunoblotting confirmatory technique. Following the administration study, a screening program involving approximately 6000 greyhound sport (mostly racing) samples was undertaken for rHuEPO. This resulted in the detection of the first rHuEPO positives in the world of canine or equine sport. In an additional case, endogenous HuEPO was detected in a sample submitted as greyhound urine. It was determined that this arose from the submission to control stewards, as greyhound urine, of a substance that was, in fact, human urine. This was a particularly welcome development as definitive confirmatory evidence of such sample switching can be difficult to obtain in the case of greyhounds. PMID- 17137527 TI - Quantification of dichloroiodomethane and bromochloroiodomethane in human blood by solid-phase microextraction coupled with gas chromatography-high-resolution mass spectrometry. AB - Iodine-containing trihalomethanes (iodo-THMs) are formed as disinfection byproducts when iodide-containing water is disinfected using chloramination process. Subsequent water use may lead to human exposure to iodo-THMs. Because of health concerns surrounding exposure to iodo-THMs, a rapid, reliable, and high throughput analytical method was developed to quantify trace levels of two iodo THMs: dichloroiodomethane (IDCM) and bromochloroiodomethane (IBCM) in human blood. These analytes from the headspace above blood samples were extracted using solid-phase microextraction. Analytes were then desorbed and separated by capillary gas chromatography and analyzed by high-resolution mass spectrometry with multiple ion monitoring. This method utilizes stable isotope dilution to quantify parts-per-trillion levels of all analytes, with excellent precision of < 9% coefficient of variation. At three spiked levels, method accuracy of IDCM and IBCM ranged between 6 and 20% difference when comparing spiked and measured amounts. The method limit of detection was 2 ng/L for both IDCM and IBCM. This selective, sensitive, and rapid method will help to assess human exposure to iodo THMs and to study potential associations between exposure and adverse health outcomes. PMID- 17137528 TI - Use of capillary electrophoresis and poly(ethylene oxide) as the coating agent for the determination of substances related to heroin addiction and treatment. AB - The purpose of this study was to identify and quantify morphine, codeine, methadone, and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine using capillary electrophoresis in urine specimens. Adequate peak separation was achieved using nearly neutral pH phosphate buffer and poly(ethylene oxide) as the coating agent. This dynamic coating of the inner surface of the capillary was obtained by rinsing with a solution containing this compound. The electroosmotic flow and the interactions between analytes and the capillary wall surface were reduced, while resolution and reproducibility were thus improved. Detection limits were appropriate for usual analytical requirements in forensic laboratories. PMID- 17137529 TI - Unexpectedly dangerous escargot stew: oleandrin poisoning through the alimentary chain. AB - A female, aged 43 and a male, aged 66, experienced gastrointestinal and cardiovascular symptoms after a meal including snail stew. Twelve hours after the ingestion, they presented with nausea, vomiting, diarrhea, and cardiovascular symptoms typical of acute toxic digoxin ingestion and were hospitalized. The man's electrocardiogram was altered, and the woman's was normal. Serum digoxin levels, measured on a Roche COBAS Integra 800 with the Roche On-Line Digoxin reagent, were 1.14 and 1.00 nmol/L, respectively. Potassium levels were normal in both patients. The serum digoxin concentration decreased on the second day, and symptoms resolved on the third day with patients fully recovered (i.e., reversion to a normal sinus rhythm). Cardiac-glycoside-like intoxication symptoms follow the ingestion of leaves or flowers of Nerium oleander. The consumed snails were suspected to be responsible for the intoxication. In the homogenized snail tissue, the concentration expressed in digoxin equivalents was 0.282 nmol/g. The presence of oleandrin and oleandrigenin in the snails was confirmed by liquid chromatography-tandem mass spectrometry analysis, which was performed on a ionic trap Finnigan LXQ instrument using an electrospray ionization interface. High pressure liquid chromatographic separation was performed on a C18 column with a gradient of methanol/water. An extract of oleander leaves was used as reference. PMID- 17137530 TI - Therapeutic and toxic concentrations of mirtazapine. AB - Six cases involving the antidepressant mirtazapine were analyzed in detail at the San Diego County Medical Examiner's Office from 2004 to 2005. Mirtazapine was initially detected and confirmed in each of these cases by a liquid-liquid gas chromatography (GC)-MS basic drug screen. Following another liquid-liquid basic extraction, mirtazapine was quantitated by GC with nitrogen-phosphorus detection. For each case, mirtazapine concentrations in peripheral blood (pb), central blood (cb), vitreous (vit), and liver were determined against matrix specific calibration curves (limit of detection 0.01 mg/L; linear range 0.025-1.0 mg/L). In contrast with earlier studies of postmortem distribution of mirtazapine, we found concentrations in liver that were significantly higher. Mirtazapine was identified in the cause of death by the pathologist in three cases. In the drug related deaths, mirtazapine concentrations (mean +/- S.D.) were 2.0 +/- 1.5 mg/L (pb), 1.6 +/- 1.0 mg/L (cb), 0.78 +/- 0.56 mg/L (vit), and 10 +/- 7.4 mg/kg (liver). Alternatively, concentrations considered therapeutic (three non-drug related deaths) were (mean +/- S.D.) 0.18 +/- 0.22 mg/L (pb), 0.16 +/- 0.17 mg/L (cb), 0.12 +/- 0.16 mg/L (vit), and 0.73 +/- 0.68 mg/kg (liver). Although mirtazapine concentrations were elevated in blood and liver in three cases, it should be noted that other drugs were also found in toxic concentrations in each case. These data may further support the fact that mirtazapine is a relatively safe drug with respect to overdose. PMID- 17137531 TI - Stability study of fluoxetine in formalin-fixed liver tissue. AB - In the present work, we report conversion of fluoxetine (Prozac), a novel anti depressant to N-methyl fluoxetine in formalin fixed liver tissue. Earlier studies indicate that drugs containing secondary amino group will react with formalin to form corresponding N-methyl derivatives. Even though embalming cadavers is common, it may create problems for forensic toxicologists if a case was not previously suspected. In formalin solutions, fluoxetine is methylated producing N methyl fluoxetine. N-Methyl fluoxetine standard was synthesized by treating fluoxetine in formaldehyde solution. The structure confirmed by (1)HNMR and gas chromatography-mass spectrometry in electron impact ionization mode. Randomly chosen rat liver pieces (200-250 mg) were injected with 100 microg of Fluoxetine. The liver pieces were covered with three different concentrations of formalin, 5%, 10%, and 20%, and at three different pHs, 3.0, 7.0, and 9.5. The reaction was studied for a total period of 30 days, and the reaction products were monitored on days 0, 4, 14, and 30 days. The study indicates that the rate of conversion of fluoxetine to its N-methyl derivative increased with increase in the concentration of formalin and pH of the solution. The conversion is rapid at higher pH values. Fluoxetine was totally converted to its N-methyl derivatives after 30 days in 20% formalin at pH 9.5. Therefore, analysis for parent drug or its N-methyl derivative in embalmed tissues may provide data that will reduce the likelihood of false negatives. PMID- 17137532 TI - Suicidal inhalation of motorbike exhaust: adding new data to the literature about the contribution of gasoline in the cause of death. AB - We would like to alert toxicologists to the importance of testing for gasoline, and for volatile hydrocarbons in general, in deaths involving inhalation of exhaust fumes occurring in closed spaces with running motors or machinery. We present here a case of suicidal inhalation of motorbike exhaust, a mixture of carbon monoxide (CO) and gasoline vapor, by a 38-year-old female. She was found in her closed home garage with a hose extending from the exhaust pipe of a motorbike through a cellophane plastic device into a closed tent in which the victim lay. She left two suicide notes nearby. The carboxyhemoglobin (COHb) was measured using visible spectrophotometry. The toxicological screening and quantitation of gasoline was performed by means of gas chromatography with flame ionization detector and confirmation was performed using gas chromatography-mass spectrometry. The %COHb determined in blood was 73%. Gasoline concentrations in heart blood and vitreous humor were 22.3 and 1.0 mg/L, respectively. Although fatalities with CO at this rate are common, we would like to highlight the role of gasoline and add new quantitative data of this toxic substance to the scarce literature. Based upon the toxicological data, along with the information provided by the medical examiner, the cause of death was determined to be CO and gasoline poisoning and the manner of death suicide. PMID- 17137533 TI - Detection of ethylglucuronide in urine following the application of Germ-X. PMID- 17137534 TI - Management of group A beta-hemolytic streptococcal pharyngotonsillitis in children. AB - Acute pharyngotonsillitis is one of the most common infections encountered by pediatricians and family physicians. According to the US Vital Health Statistics report, acute pharyngotonsillitis is responsible for more than 6 million office visits each year by children younger than 15 years of age and an additional 1.8 million visits by adolescents and young adults aged 15 to 24 years. Most children with acute pharyngotonsillitis have symptoms that can be attributed to infection with a respiratory virus, such as adenovirus, influenza virus, parainfluenza virus, rhinovirus, and respiratory syncytial virus. However, in approximately 30% to 40% of cases, acute pharyngotonsillitis is of bacterial etiology. Group A beta hemolytic streptococci (GABHS) are responsible for most bacterial cases of acute pharyngotonsillitis, although other pathogens, such as Neisseria gonorrhoeae, Arcanobacterium haemolyticum, Mycoplasma pneumoniae, and Chlamydia pneumoniae, may be the causative agents in sporadic cases. Pharyngotonsillitis caused by these latter pathogens can sometimes be distinguished from that caused by GABHS by considering the patient's medical history in concert with the clinical presentation. In some cases, acute pharyngotonsillitis may have an idiopathic etiology. An accurate diagnosis of GABHS infection is important because it is the only common form of acute pharyngotonsillitis for which antibiotic therapy is definitely indicated. Antibiotic therapy can shorten the clinical course of GABHS pharyngotonsillitis, reduce the rate of transmission, and prevent suppurative and nonsuppurative complications, such as peritonsillar abscess and acute rheumatic fever. Although the threat of rheumatic fever is much lower for children in the United States than in developing nations, preventing rheumatic fever and the spread of disease is the primary goal of antibiotic therapy in GABHS pharyngotonsillitis treatment and a cornerstone of practice guidelines. PMID- 17137539 TI - Migraine: a better way to recognize and treat it. AB - Consider using the Headache Assessment Quiz, which 76% of providers in this study said enabled patients to adequately convey headache severity/symptoms, compared with just 20% of providers at baseline who thought patients communicated clearly. Use the quiz also to better understand the impact of migraine on a patient's life, and to help determine which patients need migraine-specific therapy. PMID- 17137540 TI - Hyperpigmentation and vesicles after beach vacation. Phytophotodermatitis. PMID- 17137541 TI - The preteen visit: an opportunity for prevention. AB - All early adolescents should visit a physician at age 11 or 12 years to receive a set of recommended vaccines. Two vaccines are recommended for boys in this age group-quadrivalent meningococcal conjugate vaccine (MCV4) and tetanus toxoid, reduced diphtheria, and acellular pertussis vaccine (Tdap). Three vaccines are recommended for girls--MCV4, Tdap, and human papilloma virus (HPV) vaccine. In addition, 2 doses of varicella vaccine are now recommended before age 5 years; both boys and girls at age 11 or 12 who have received only 1 dose should be given a second. PMID- 17137542 TI - Do family physicians fail to provide triptans for patients with migraine? AB - While continuing to improve recognition of migraine in your patient population, pay particular attention to the adherence rate among those for whom you have prescribed a triptan. Ask patients who discontinue triptan therapy why they made that decision. Besides adverse effects from the agent, reasons may include medication cost, influence of comorbidities, or triptan interaction with medications you may not have known about. PMID- 17137543 TI - Is any one analgesic superior for episodic tension-type headache? AB - Though all non-narcotic analgesics have equivalent efficacy against tension-type headache, ibuprofen's generally favorable side-effect profile makes it a reasonable first choice. PMID- 17137544 TI - Emergency contraception care. AB - This guideline targets women who have had unprotected or inadequately protected intercourse within the past 120 hours and do not desire pregnancy. Practitioners can make informed decisions about obstetric and gynecologic care, given the evidence in this guideline regarding safety, efficacy, risks and benefits of the use of emergency contraception including progestin-only and combined estrogen progestin regimen. The major outcome considered was incidence of unintended pregnancy. The evidence rating is updated to comply with the SORT taxonomy. PMID- 17137545 TI - Clinical inquiries: What is the dietary treatment for low HDL cholesterol? AB - Low-carbohydrate diets raise high-density lipoprotein (HDL) cholesterol levels by approximately 10%; soy protein with isoflavones raises HDL by 3% (strength of recommendation [SOR]: C, based on meta-analysis of physiologic parameters). The Dietary Approaches to Stop Hypertension (DASH) diet and multivitamin supplementation raise HDL 21% to 33% (SOR: C, based on single randomized trial each measuring physiologic parameters). No other dietary interventions studied raise HDL (SOR: C, based on meta-analysis of physiologic parameters). PMID- 17137546 TI - Clinical inquiries: When are antibiotics indicated for acute COPD exacerbations? AB - Antibiotics (including those given orally) reduce mortality and treatment failures for hospitalized patients with acute exacerbations of chronic obstructive pulmonary disease (COPD) (strength of recommendation [SOR]: A, based on systematic reviews). Antibiotics may be prescribed in the outpatient setting for those with severe exacerbations (SOR: C, based on expert opinion). PMID- 17137547 TI - Clinical inquiries: How should you manage a depressed patient unresponsive to an SSRI? AB - The best approach among studied alternatives to manage a patient with treatment resistant depression is not clear from the evidence. All of the options reviewed seem to have about a 25% to 30% success rate. Switching to other antidepressants or augmenting with non-antidepressant drugs has the best supporting evidence (strength of recommendation [SOR]: B).1 Adding additional antidepressants (SOR: B), using psychotherapy (SOR: B), and initiating electroconvulsive therapy (ECT) (SOR: C) are options. Various antidepressants are used as add-on therapy. Psychotherapy is often recommended, though the evidence of benefit after a failed course of initial therapy is sparse. The evidence supporting use of ECT in treatment-resistant depression is weak. Comparison among the options is based on expert opinion (SOR: C). Additional reports from the STAR*D trial may improve the quality of the evidence in the near future. PMID- 17137548 TI - Clinical inquiries: What are safe sleeping arrangements for infants? AB - Non-supine sleep position and parental tobacco use are known risk factors for sudden infant death syndrome (SIDS). Recent studies show that co-sleeping (bed sharing) slightly increases the overall risk of SIDS (strength of recommendation [SOR]: B) and is greatest for infants less than 11 weeks old (SOR: B). The relationship between bed sharing and SIDS is strongest for infants whose parents use tobacco (SOR: B). Infants who sleep in a room separate from their caregivers or on a couch or an armchair are at increased risk for SIDS (SOR: B). Using bedding accessories such as duvets or pillows may increase an infant's risk of SIDS (SOR: B). PMID- 17137549 TI - Clinical inquiries: What blood tests help diagnose celiac disease? AB - Histological confirmation of infiltrative lesions via small bowel biopsy is the gold standard for diagnosing celiac disease. Four serum antibody assays may serve as a first-step diagnostic tool to identify biopsy candidates: immunoglobulin A tissue transglutaminase (IgA tTG), IgA endomysial antibody (IgA EMA), IgA antigliadin antibody (IgA AGA), and IgG antigliadin antibody (IgG AGA). IgA tTG and IgA EMA offer the best diagnostic accuracy. Patients with selective IgA deficiency may have falsely negative IgA assays (strength of recommendation [SOR]: B, based on a systematic review, multiple small cross-sectional studies, and expert opinion). PMID- 17137550 TI - Clinical inquiries: Do glucosamine and chondroitin worsen blood sugar control in diabetes? AB - Despite theoretical risks based on animal models given high intravenous doses, glucosamine/chondroitin (1500 mg/1200 mg daily) does not adversely affect short term glycemic control for patients whose diabetes is well-controlled, or for those without diabetes or glucose intolerance (SOR: A, consistent, good-quality patient-oriented evidence). Some preliminary evidence suggests that glucosamine may worsen glucose intolerance for patients with untreated or undiagnosed glucose intolerance or diabetes (SOR: C, extrapolation from disease-oriented evidence). Long-term effects are unknown; however, no compelling theoretical or incidental data suggest that long-term results should be different (SOR: C, expert opinion). Further studies are required to clarify the effects of glucosamine on patients with poorly controlled diabetes or glucose intolerance. PMID- 17137551 TI - Same-sample analysis of ethoxyresorufin-O-deethylase activity and cytochrome P4501A mRNA abundance in chicken embryo hepatocytes. AB - Inducibility of the cytochrome P4501A4 (CYP1A4) enzyme, measured as ethoxyresorufin-O-deethylase (EROD) activity, has been used as a biomarker for sensitivity to the effects of dioxin-like compounds in avian species. Here, we present a quantitative reverse transcriptase-PCR (Q-PCR) method for assessing this biomarker response at the level of messenger RNA (mRNA) expression. The method was validated for use in fresh samples as well as samples that have been analyzed for EROD activity previously. Concentration-dependent effects of 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) on CYP1A4 and CYP1A5 mRNA abundance were detected in fresh and post-EROD hepatocyte cultures. Although the quality of the RNA obtained from post-EROD samples was low, quantification of the CYP1A mRNA response to TCDD was not compromised. Several benefits of evaluating CYP1A mRNA expression in addition to EROD activity were noted. The CYP1A mRNA bioassay may provide more accurate estimates for the potency of environmental mixtures of contaminants and has a very low detection limit. When working with hepatocytes cultured from wild or endangered species, our approach can help to circumvent the problem of small sample size by maximizing the amount of data obtained from each sample. PMID- 17137552 TI - Functional significance of strain distribution in the human mandible under masticatory load: numerical predictions. AB - A common feature of studies of mandibular morphology is the assumption that there is some functional relation between the form of the lower jaw and masticatory stress. It was noted that the local variation in cortical bone thickness in the mandibular corpus appears to be stereotypical among anthropoids. This occurs at sections under the molars, where the lingual cortical plate is thinner than buccal one. In this study we investigate and contrast the strain pattern along buccal and lingual surfaces of the mandibular corpus during mastication using a numerical model of a human mandible. We show that strain distribution differs in alveolar and mid-corpus segments of the mandible and that the latter develops an alternate pattern between the buccal and lingual aspects of the working and balancing sides of the jaw. We then relate the magnitude of these strains to Frost's mechanostat. Our results suggest that the cortical asymmetry of the human mandible is in fact not related to strain patterns generated during mastication. PMID- 17137553 TI - Therapeutic effect of boron neutron capture therapy (BNCT) on field cancerized tissue: inhibition of DNA synthesis and lag in the development of second primary tumors in precancerous tissue around treated tumors in DMBA-induced carcinogenesis in the hamster cheek pouch oral cancer model. AB - OBJECTIVE: We previously reported the therapeutic success of different BNCT protocols in the treatment of oral cancer, employing the hamster cheek pouch model. The aim of the present study was to evaluate the effect of these BNCT protocols on DNA synthesis in precancerous and normal tissue in this model and assess the potential lag in the development of second primary tumors in precancerous tissue. The data are relevant to potential control of field cancerized tissue and tolerance of normal tissue. MATERIALS AND METHODS: We evaluated DNA synthesis in precancerous and normal pouch tissue 1-30 days post BNCT mediated by boronophenylalanine (BPA), GB-10 (Na(2)(10)B(10)H(10)) or (BPA+GB-10) employing incorporation of 5-bromo-2'-deoxyuridine as an end-point. The BNCT-induced potential lag in the development of second primary tumors from precancerous tissue was monitored. RESULTS: A drastic, statistically significant reduction in DNA synthesis occurred in precancerous tissue as early as 1 day post BNCT and was sustained at virtually all time-points until 30 days post-BNCT for all the protocols. The histological categories evaluated individually within precancerous tissue (dysplasia, hyperplasia and NUMF [no unusual microscopic features]) responded similarly. DNA synthesis in normal tissue treated with BNCT oscillated around the very low pre-treatment values. A BNCT-induced lag in the development of second primary tumors was observed. CONCLUSIONS: BNCT induced a drastic fall in DNA synthesis in precancerous tissue that would be associated to the observed lag in the development of second primary tumors. The minimum variations in DNA synthesis in BNCT-treated normal tissue would correlate with the absence of normal tissue radiotoxicity. The present data would support the control of field-cancerized areas by BNCT. PMID- 17137554 TI - The human non-gastric H,K-ATPase has a different cation specificity than the rat enzyme. AB - The primary sequence of non-gastric H,K-ATPase differs much more between species than that of Na,K-ATPase or gastric H,K-ATPase. To investigate whether this causes species-dependent differences in enzymatic properties, we co-expressed the catalytic subunit of human non-gastric H,K-ATPase in Sf9 cells with the beta(1) subunit of rat Na,K-ATPase and compared its properties with those of the rat enzyme (Swarts et al., J. Biol. Chem. 280, 33115-33122, 2005). Maximal ATPase activity was obtained with NH(4)(+) as activating cation. The enzyme was also stimulated by Na(+), but in contrast to the rat enzyme, hardly by K(+). SCH 28080 inhibited the NH(4)(+)-stimulated activity of the human enzyme much more potently than that of the rat enzyme. The steady-state phosphorylation level of the human enzyme decreased with increasing pH, [K(+)], and [Na(+)] and nearly doubled in the presence of oligomycin. Oligomycin increased the sensitivity of the phosphorylated intermediate to ADP, demonstrating that it inhibited the conversion of E(1)P to E(2)P. All three cations stimulated the dephosphorylation rate dose-dependently. Our studies support a role of the human enzyme in H(+)/Na(+) and/or H(+)/NH(4)(+) transport but not in Na(+)/K(+) transport. PMID- 17137555 TI - Proteomics-based identification of biomarkers for predicting sensitivity to a PI3 kinase inhibitor in cancer. AB - To identify biomarkers for predicting sensitivity to phosphatidylinositol 3 kinase (PI3K) inhibitors, we have developed a proteomics-based approach. Using surface-enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF MS), we measured the expression of 393 proteins in 39 human cancer cell lines (JFCR-39), and combined it with our previously established chemosensitivity database to select for proteins whose expressions show significant correlations to drug sensitivities. This integrated approach allowed us to identify peaks from two proteins, 11.6 and 11.8 kDa, that showed significant correlations with the sensitivity to a PI3K inhibitor, LY294002. We found that the 11.8 kDa protein was a phosphorylated form of the 11.6 kDa protein. While the 11.8 kDa protein showed a positive correlation with the sensitivity to LY294002, the 11.6 kDa protein showed a negative correlation with that of the LY294002. The 11.6 kDa protein was purified chromatographically, and was identified by SELDI-TOF MS as the ribosomal P2 protein, which possesses two prospective phosphorylation sites. These results suggested that the phosphorylation status of the ribosomal P2 was responsible for determining the sensitivity to LY294002, and that the ribosomal P2 could be a potential biomarker for predicting chemosensitivity. PMID- 17137556 TI - Pharmacological inhibition of the MAPK/ERK pathway increases sensitivity to 2 chloro-2'-deoxyadenosine (CdA) in the B-cell leukemia cell line EHEB. AB - EHEB leukemic cells, which are derived from a patient suffering B-cell chronic lymphocytic leukemia (B-CLL), display intermediate sensitivity to the purine analogue 2-chloro-2'-deoxyadenosine (CdA). Because the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathway can rescue cancer cells from apoptotic signals, we investigated MAPK/ERK signaling in EHEB cells in response to CdA. We observed that CdA, at concentrations around its IC50, dose- and time-dependently increased the phosphorylation state of ERK 1/2 (p-ERK), indicating an activation of the MAPK/ERK pathway. This activation required CdA metabolism and de novo protein synthesis, and was independent on caspase activation. Interruption of ERK signaling, using the specific MEK inhibitors U-0126 and PD-98059, significantly enhanced CdA cytotoxicity, evaluated by the MTT assay. Drug interaction analysis showed synergism in the majority of combinations between CdA and MEK inhibitors tested. MEK inhibitors also dramatically increased apoptosis induced by CdA alone, evaluated by caspase 3 activation and poly (ADP-ribose) polymerase (PARP) cleavage. Collectively, these observations show that ERK 1/2 activation elicited by CdA serves as a cytoprotective function and suggest that inhibitors of this pathway could be combined with CdA in the treatment of selected hematological malignancies. PMID- 17137557 TI - Gene expression and regulation of drug transporters in the intestine and kidney. AB - Intestinal absorption and renal secretion of ionic drugs are controlled by a number of drug transporters expressed at the brush-border and basolateral membranes of epithelial cells. Over the last several years, considerable progress has been made regarding the molecular identification and functional characterization of drug transporters. Under some physiological and pathophysiological conditions, the expression and transport activity of drug transporters are changed, affecting the pharmacokinetics of substrate drugs. The regulation of transport activity in response to endogenous and exogenous signals can occur at various levels such as transcription, mRNA stability, translation, and posttranslational modification. Transcriptional regulation is of particular interest, because changes in transport activity are dynamically regulated by increases or decreases in levels of mRNA expression. The tissue-specific expression of drug transporters is also under transcriptional control, and recent studies using clinical samples from human tissues have revealed the expression profiles of drug transporters in the human body. The purpose of this research updates is to review the recent progress in the study of the gene expression and regulation of intestinal and renal drug transporters. PMID- 17137558 TI - Inhibitory control in high-functioning autism: decreased activation and underconnectivity in inhibition networks. AB - BACKGROUND: Inhibiting prepotent responses is critical to optimal cognitive and behavioral function across many domains. Several behavioral studies have investigated response inhibition in autism, and the findings varied according to the components involved in inhibition. There has been only one published functional magnetic resonance imaging (fMRI) study so far on inhibition in autism, which found greater activation in participants with autism than control participants. METHODS: This study investigated the neural basis of response inhibition in 12 high-functioning adults with autism and 12 age- and intelligence quotient (IQ)-matched control participants during a simple response inhibition task and an inhibition task involving working memory. RESULTS: In both inhibition tasks, the participants with autism showed less brain activation than control participants in areas often found to be active in response inhibition tasks, namely the anterior cingulate cortex. In the more demanding inhibition condition, involving working memory, the participants with autism showed more activation than control participants in the premotor areas. In addition to the activation differences, the participants with autism showed lower levels of synchronization between the inhibition network (anterior cingulate gyrus, middle cingulate gyrus, and insula) and the right middle and inferior frontal and right inferior parietal regions. CONCLUSIONS: The results indicate that the inhibition circuitry in the autism group is activated atypically and is less synchronized, leaving inhibition to be accomplished by strategic control rather than automatically. At the behavioral level, there was no difference between the groups. PMID- 17137559 TI - Hypothalamic-pituitary-adrenal axis function in dissociative disorders, post traumatic stress disorder, and healthy volunteers. AB - BACKGROUND: This study investigated basal and stress-induced hypothalamic pituitary-adrenal (HPA)-axis alterations in dissociative disorders (DDs). METHODS: Forty-six subjects with DD without lifetime post-traumatic stress disorder (PTSD), 35 subjects with PTSD, and 58 healthy comparison (HC) subjects, free of current major depression, were studied as inpatients. After a 24-hour urine collection and hourly blood sampling for ambient cortisol determination, a low-dose dexamethasone suppression test was administered, followed by the Trier Social Stress Test. RESULTS: The DD group had significantly elevated urinary cortisol compared with the HC group, which was more pronounced in the absence of lifetime major depression, whereas the PTSD and HC groups did not differ. The DD group demonstrated significantly greater resistance to, and faster escape from, dexamethasone suppression compared with the HC group, whereas the PTSD and HC groups did not differ. The three groups did not differ in cortisol stress reactivity, but both psychiatric groups demonstrated a significant inverse correlation between dissociation severity and cortisol reactivity, after controlling for all other symptomatology. The PTSD subgroup with comorbid DD tended to have blunted stress reactivity compared with the HC group. CONCLUSIONS: The study demonstrates a distinct pattern of HPA-axis dysregulation in DDs, emphasizing the importance of further study of stress-response systems in dissociative psychopathology. PMID- 17137560 TI - Efficacy and safety of dexmethylphenidate extended-release capsules in adults with attention-deficit/hyperactivity disorder. AB - BACKGROUND: This multicenter, randomized, fixed-dose, double-blind, placebo controlled study evaluated efficacy of extended-release dexmethylphenidate (d-MPH ER) in adults with attention-deficit/hyperactivity disorder (ADHD). METHODS: Randomized adults with ADHD (n=221) received once-daily d-MPH-ER 20 mg, 30 mg, or 40 mg or placebo for 5 weeks. The primary efficacy variable was change from baseline to final visit in DSM-IV ADHD Rating Scale (ADHD-RS) total score. Secondary efficacy parameters included the proportion of patients with improvement>or=30% in ADHD-RS total score and final scores on Clinical Global Impressions-Improvement (CGI-I) scale. RESULTS: Of 218 evaluable patients, 184 completed the study. All d-MPH-ER doses were significantly superior to placebo in improving ADHD-RS total scores. Placebo scores improved by 7.9; d-MPH-ER, 20 mg, improved by 13.7 (p=.006); d-MPH-ER, 30 mg, improved by 13.4 (p=.012); and d-MPH ER, 40 mg, improved by 16.9 (p<.001). Overall distribution of CGI-I ratings at final visit was significantly better with each d-MPH-ER dosage than with placebo. There were no unexpected safety or tolerability concerns, based on experience with racemic methylphenidate (MPH) in adults and dexmethylphenidate (d-MPH) in children. CONCLUSIONS: Once-daily d-MPH-ER at 20 mg, 30 mg, or 40 mg is a safe and effective treatment for adults with ADHD. PMID- 17137561 TI - Oxytocin improves "mind-reading" in humans. AB - BACKGROUND: The ability to "read the mind" of other individuals, that is, to infer their mental state by interpreting subtle social cues, is indispensable in human social interaction. The neuropeptide oxytocin plays a central role in social approach behavior in nonhuman mammals. METHODS: In a double-blind, placebo controlled, within-subject design, 30 healthy male volunteers were tested for their ability to infer the affective mental state of others using the Reading the Mind in the Eyes Test (RMET) after intranasal administration of 24 IU oxytocin. RESULTS: Oxytocin improved performance on the RMET compared with placebo. This effect was pronounced for difficult compared with easy items. CONCLUSIONS: Our data suggest that oxytocin improves the ability to infer the mental state of others from social cues of the eye region. Oxytocin might play a role in the pathogenesis of autism spectrum disorder, which is characterized by severe social impairment. PMID- 17137562 TI - Intrapair differences in hippocampal volume in monozygotic twins discordant for the risk for anxiety and depression. AB - BACKGROUND: Current biological psychiatric models assume that genetic and environmental risk factors for anxiety and depression act on the same brain structures. METHODS: To test this assumption, we assessed brain anatomy by using optimized voxel-based morphometry on magnetic resonance images obtained in monozygotic twin pairs who were discordant for the risk of anxiety and depression (n = 10 pairs) and in monozygotic twin pairs who were concordant for high (n = 7 pairs) or low (n = 15 pairs) risk for anxiety and depression. RESULTS: We observed volume reductions in the temporal lobe, most notably in the left posterior hippocampal region in subjects at high risk for anxiety and depression, but exclusively in the intrapair comparison of discordant monozygotic twins. Because monozygotic twins are genetically identical, any discordance in their risk for anxiety and depression and hippocampal volume must arise from differential exposure to environmental influences. A group comparison between pairs concordant for low or high risk, which is more likely to reflect differences in genetic vulnerability, did not show reduced temporal-lobe and posterior hippocampal volumes in the pairs at high risk for anxiety and depression. CONCLUSIONS: This pattern of results suggests that damage to temporal lobe structures may be specific to an environmentally driven etiology of anxiety and depression. PMID- 17137563 TI - Understanding genetic risk for aggression: clues from the brain's response to social exclusion. AB - BACKGROUND: Although research indicates a relationship between the monoamine oxidase-A (MAOA) gene and aggression, the intervening neural and psychological mechanisms are unknown. Individuals with the low expression allele (MAOA-L) of a functional polymorphism in the MAOA gene might be prone to aggression because they are socially or emotionally hyposensitive and thus care less about harming others or because they are socially or emotionally hypersensitive and thus respond to negative social experiences with defensively aggressive behavior. METHODS: We investigated the relationships between the MAOA polymorphism, trait aggression, trait interpersonal hypersensitivity, and neural responses to social exclusion in 32 healthy men and women. RESULTS: The MAOA-L individuals (men and women) reported higher trait aggression than individuals with the high expression allele (MAOA-H). The MAOA-L individuals reported higher trait interpersonal hypersensitivity and showed greater dorsal anterior cingulate cortex (dACC) activity (associated with rejection-related distress) to social exclusion compared with MAOA-H individuals, consistent with a social hypersensitivity hypothesis. Moreover, the MAOA-aggression relationship was mediated by greater dACC reactivity to social exclusion, suggesting that MAOA might relate to aggression through socioemotional hypersensitivity. CONCLUSIONS: These data suggest that the relationship between MAOA and aggression might be due to a heightened rather than a reduced sensitivity to negative socioemotional experiences like social rejection. PMID- 17137565 TI - Salivary cortisol and psychopathology in children bereaved by the september 11, 2001 terror attacks. AB - BACKGROUND: Studies suggest that stressful events increase risk for childhood anxiety and depression and hypothalamic-pituitary-adrenal (HPA) axis dysregulation. This prospective longitudinal study evaluated relationships among severe psychosocial stress, psychiatric morbidity, and HPA axis function in children. METHODS: Forty-five children (mean age: 8.9 +/- 2.9 years) suffering parent death from September 11, 2001 terror attacks and 34 nonbereaved children (mean age: 9.3 +/- 2.5 years) were evaluated prospectively at 6-month intervals in this 2-year study. Assessments involved diagnostic interviews (Child Schedule for Affective Disorders and Schizophrenia [K-SADS]) for psychopathology and 3 days of baseline salivary cortisol and a salivary dexamethasone suppression test for HPA axis function. RESULTS: Bereaved children, but not nonbereaved children, had significantly increased rates of psychiatric disorders involving anxiety disorders, especially posttraumatic stress disorder (PTSD), after September 11, 2001 compared with retrospective assessments before September 11, 2001. Morning (AM) and 4:00 pm baseline cortisol were significantly and persistently higher for bereaved than nonbereaved children. Compared with bereaved children without psychopathology, bereaved children with PTSD had significantly lower 4:00 pm baseline cortisol and significantly greater 4:00 pm cortisol suppression. Children with generalized anxiety disorder had significantly less AM cortisol suppression than children without psychopathology. CONCLUSIONS: Children bereaved by sudden, unexpected parent death had persistent psychological dysfunction and HPA axis dysregulation in this study. PMID- 17137564 TI - Rate of head growth decelerates and symptoms worsen in the second year of life in autism. AB - BACKGROUND: Longitudinal studies of head circumference growth in infants later diagnosed with autism are needed to understand the accelerated head growth in this disorder. METHODS: We analyzed longitudinal head circumference data from birth to 3 years in 28 children later diagnosed with autism spectrum disorder on the basis of individual growth curve analyses using hierarchical linear models. RESULTS: Head circumference Z scores relative to norms significantly increased in the autism sample from birth to 12 months, but this pattern did not persist beyond 12 months. Rather, the rate of change in head circumference from 12 to 36 months was not different from the normative sample. CONCLUSIONS: These results suggest that a period of exceptionally rapid head growth occurs during the first year of life in autism; after 12 months of age, the rate of head circumference growth decelerates relative to the rate during the first year of life. Studies of behavioral development in infants later diagnosed with autism suggest that the period of acceleration of head growth precedes and overlaps with the onset of behavioral symptoms, and the period of deceleration coincides with a period of worsening of symptoms in the second year of life. PMID- 17137566 TI - Genetic deficiency of cyclooxygenase-2 attenuates abdominal aortic aneurysm formation in mice. AB - OBJECTIVE: Abdominal aortic aneurysms (AAAs) are characterized by chronic inflammation which contributes to the remodeling and eventual weakening of the vessel wall. Increased cyclooxygenase-2 (COX-2) expression is detected in human aneurysmal tissue and is suggested to contribute to the disease. The aim of the current study was to define the role of COX-2 expression in the development of AAAs, using a model of the disease. METHODS: AAAs were induced in mice by chronic angiotensin II infusion, and were analyzed following 3, 7, 21 or 28 days of the infusion. AAA incidence and severity, together with the expression of inflammatory markers, were compared between abdominal aortas from COX-2-deficient mice and their wild-type littermate controls. RESULTS: The AAA incidence in COX-2 wild-type mice was 54% (13/24), whereas AAAs were not detected in COX-2-deficient mice (0/23) following 28 days of angiotensin II infusion. The genetic deficiency of COX-2 also resulted in a 73% and 90% reduction in AAA incidence following 7 and 21 days of angiotensin II infusion, respectively. In COX-2 wild-type mice, COX-2 mRNA expression in the abdominal aorta was induced by angiotensin II beginning 3 days following initiation of the infusion, which continued throughout progression of the disease. Abundant COX-2 protein expression was detected in medial smooth muscle cells adjacent to the AAAs. The deficiency of COX-2 significantly attenuated mRNA expression in the abdominal aorta of the macrophage marker CD68, and the inflammatory cell recruitment chemokines, monocyte chemotactic protein-1 and macrophage inflammatory protein-1alpha. CONCLUSIONS: Our findings suggest that increased COX-2 expression in smooth muscle cells of the abdominal aorta contributes to AAA formation in mice by enhancing inflammatory cell infiltration. PMID- 17137567 TI - Does the adenosine A2A receptor stimulate the ryanodine receptor? PMID- 17137568 TI - Tissue kallikrein protects against pressure overload-induced cardiac hypertrophy through kinin B2 receptor and glycogen synthase kinase-3beta activation. AB - OBJECTIVE: We assessed the role of glycogen synthase kinase-3beta (GSK-3beta) and kinin B2 receptor in mediating tissue kallikrein's protective effects against cardiac hypertrophy. METHODS: We investigated the effect and mechanisms of tissue kallikrein using hypertrophic animal models of rats as well as mice deficient in kinin B1 or B2 receptor after aortic constriction (AC). RESULTS: Intramyocardial delivery of adenovirus containing the human tissue kallikrein gene resulted in expression of recombinant kallikrein in rat myocardium. Kallikrein gene delivery improved cardiac function and reduced heart weight/body weight ratio and cardiomyocyte size without affecting mean arterial pressure 28 days after AC. Icatibant and adenovirus carrying a catalytically inactive GSK-3beta mutant (Ad.GSK-3beta-KM) abolished kallikrein's effects. Kallikrein treatment increased cardiac nitric oxide (NO) levels and reduced NAD(P)H oxidase activity and superoxide production. Furthermore, kallikrein reduced the phosphorylation of apoptosis signal-regulating kinase1, mitogen-activated protein kinases (MAPKs), Akt, GSK-3beta, and cAMP-response element binding (CREB) protein, and decreased nuclear factor-kappaB (NF-kappaB) activation in the myocardium. Ad.GSK-3beta-KM abrogated kallikrein's actions on GSK-3beta and CREB phosphorylation and NF kappaB activation, whereas icatibant blocked all kallikrein's effects. The protective role of kinin B2 receptor in cardiac hypertrophy was further confirmed in kinin receptor knockout mice as heart weight/body weight ratio and cardiomyocyte size increased significantly in kinin B2 receptor knockout mice after AC compared to wild type and B1 receptor knockout mice. CONCLUSIONS: These findings indicate that tissue kallikrein, through kinin B2 receptor and GSK-3beta signaling, protects against pressure overload-induced cardiomyocyte hypertrophy by increased NO formation and oxidative stress-induced Akt-GSK-3beta-mediated signaling events, MAPK and NF-kappaB activation. PMID- 17137569 TI - Calmodulin and CaMKII as molecular switches for cardiac ion channels. AB - Because changes in intracellular Ca(2+) concentration are the final signals of electrical activity in excitable cells, many mechanisms have evolved to regulate Ca(2+) influx. Among the most important are those pathways that directly regulate the ion channels responsible for regulating and generating the Ca(2+) influx signal. Recent work has demonstrated that the Ca(2+) binding protein calmodulin (CaM) and the Ca(2+)/CaM-sensitive kinase CaMKII are important modulators of cardiac ion channels. Thus, Ca(2+) participates in feedback modulation to control electrical activity. This review highlights various mechanisms by which CaM and CaMKII regulate cardiovascular ion channel activity and presents a novel model for CaMKII regulation of Ca(V)1.2 Ca(2+) channel function. PMID- 17137570 TI - The HMG-box protein Lilliputian is required for Runt-dependent activation of the pair-rule gene fushi-tarazu. AB - lilliputian (lilli), the sole Drosophila member of the FMR2/AF4 (Fragile X Mental Retardation/Acute Lymphoblastic Leukemia) family of transcription factors, is widely expressed with roles in segmentation, cellularization, and gastrulation during early embryogenesis with additional distinct roles at later stages of embryonic and postembryonic development. We identified lilli in a genetic screen based on the suppression of a lethal phenotype that is associated with ectopic expression of the transcription factor encoded by the segmentation gene runt in the blastoderm embryo. In contrast to other factors identified by this screen, lilli appears to have no role in mediating either the establishment or maintenance of engrailed (en) repression by Runt. Instead, we find that Lilli plays a critical role in the Runt-dependent activation of the pair-rule segmentation gene fushi-tarazu (ftz). The requirement for lilli is distinct from and temporally precedes the Runt-dependent activation of ftz that is mediated by the orphan nuclear receptor protein Ftz-F1. We further describe a role for lilli in the activation of Sex-lethal (Sxl), an early target of Runt in the sex determination pathway. However, lilli is not required for all targets that are activated by Runt and appears to have no role in activation of sloppy paired (slp1). Based on these results we suggest that Lilli plays an architectural role in facilitating transcriptional activation that depends both on the target gene and the developmental context. PMID- 17137571 TI - Compartmentalization of a unique ADP/ATP carrier protein SFEC (Sperm Flagellar Energy Carrier, AAC4) with glycolytic enzymes in the fibrous sheath of the human sperm flagellar principal piece. AB - The longest part of the sperm flagellum, the principal piece, contains the fibrous sheath, a cytoskeletal element unique to spermiogenesis. We performed mass spectrometry proteomics on isolated human fibrous sheaths identifying a unique ADP/ATP carrier protein, SFEC [AAC4], seven glycolytic enzymes previously unreported in the human sperm fibrous sheath, and sorbitol dehydrogenase. SFEC, pyruvate kinase and aldolase were co-localized by immunofluorescence to the principal piece. A homology model constructed for SFEC predicted unique residues at the entrance to the nucleotide binding pocket of SFEC that are absent in other human ADP/ATP carriers, suggesting opportunities for selective drug targeting. This study provides the first evidence of a role for an ADP/ATP carrier family member in glycolysis. The co-localization of SFEC and glycolytic enzymes in the fibrous sheath supports a growing literature that the principal piece of the flagellum is capable of generating and regulating ATP independently from mitochondrial oxidation in the mid-piece. A model is proposed that the fibrous sheath represents a highly ordered complex, analogous to the electron transport chain, in which adjacent enzymes in the glycolytic pathway are assembled to permit efficient flux of energy substrates and products with SFEC serving to mediate energy generating and energy consuming processes in the distal flagellum, possibly as a nucleotide shuttle between flagellar glycolysis, protein phosphorylation and mechanisms of motility. PMID- 17137573 TI - Role of the hindbrain in patterning the otic vesicle: a study of the zebrafish vhnf1 mutant. AB - The vertebrate inner ear develops from an ectodermal placode adjacent to rhombomeres 4 to 6 of the segmented hindbrain. The placode then transforms into a vesicle and becomes regionalised along its anteroposterior, dorsoventral and mediolateral axes. To investigate the role of hindbrain signals in instructing otic vesicle regionalisation, we analysed ear development in zebrafish mutants for vhnf1, a gene expressed in the caudal hindbrain during otic induction and regionalisation. We show that, in vhnf1 homozygous embryos, the patterning of the otic vesicle is affected along both the anteroposterior and dorsoventral axes. First, anterior gene expression domains are either expanded along the whole anteroposterior axis of the vesicle or duplicated in the posterior region. Second, the dorsal domain is severely reduced, and cell groups normally located ventrally are shifted dorsally, sometimes forming a single dorsal patch along the whole AP extent of the otic vesicle. Third, and probably as a consequence, the size and organization of the sensory and neurogenic epithelia are disturbed. These results demonstrate that, in zebrafish, signals from the hindbrain control the patterning of the otic vesicle, not only along the anteroposterior axis, but also, as in amniotes, along the dorsoventral axis. They suggest that, despite the evolution of inner ear structure and function, some of the mechanisms underlying the regionalisation of the otic vesicle in fish and amniotes have been conserved. PMID- 17137572 TI - Sine oculis, a member of the SIX family of transcription factors, directs eye formation. AB - The initiation of eye formation in all seeing animals is controlled by a group of selector genes that together forms the retinal determination cascade. In Drosophila, mice and humans, loss-of-function mutations lead to defects in eye and/or head development. While ectopic expression of these genes is sufficient to direct non-retinal tissues towards an eye fate, the ability of each gene to initiate eye formation is neither unlimited nor equal. A particularly enigmatic observation has been that one member of the cascade, sine oculis (so), which is a member of the SIX family of homeobox transcription factors, is unable to initiate eye development in non-retinal tissues. It is in contrast to every other retinal determination gene including optix, another Six family member, which can induce eye formation when expressed on its own. Here we demonstrate that, in contrast to published reports, expression of so on its own is sufficient to induce eye development within non-retinal tissues. We have extended results from prior reports on binding partner selectivity and DNA binding sites by conducting a structure/function analysis of the SO and OPTIX proteins. Here we demonstrate that the SIX domains and C-terminal portions of the SO and OPTIX proteins are required for functional specificity of SIX class transcription factors while the homeodomain of these proteins are interchangeable. Taken together, these results shed new light on the role that so plays in eye specification. PMID- 17137574 TI - The role of nitric oxide within the nucleus accumbens on the acquisition and expression of morphine-induced place preference in morphine sensitized rats. AB - In the present study, the effects of intra-accumbal administration of L-arginine, a nitric oxide precursor, and N(G)-nitro-L-arginine methyl-ester (L-NAME), a nitric oxide synthase inhibitor, on the acquisition and expression of morphine induced place conditioning in morphine-sensitized rats were studied. Subcutaneous (s.c.) administration of morphine (2.5, 5 and 7.5 mg/kg) induced conditioned place preference. Repeated pretreatment of morphine (5 mg/kg, i.p.) followed by 5 days without drug treatment, increased conditioning response induced by morphine (0.25, 0.5 and 0.75 mg/kg). Intra-accumbal (intra-nucleus accumbens; 1 microg/rat) administration of L-arginine (0.3, 1 and 3 microg/rat) significantly increased or reduced the acquisition of morphine place conditioning in non sensitized and sensitized rats respectively. However, the drug reduced expression of place conditioning by morphine in sensitized animals. Intra-nucleus accumbens injections of L-NAME (0.3, 1 and 3 microg/rat) reduced the acquisition and expression of morphine place conditioning in the sensitized animals. The results indicate that nitric oxide (NO) within the nucleus accumbens is involved in the acquisition and expression of morphine place conditioning in morphine-sensitized rats. PMID- 17137575 TI - Characterization of ZNF23, a KRAB-containing protein that is downregulated in human cancers and inhibits cell cycle progression. AB - The Krupple-associated box-containing zinc-finger proteins (KRAB-ZFPs) make up one of the largest family of transcription factors. Several members of the KRAB ZFPs modulate cell growth, survival and are implicated in malignant disorders. However, most members are not well characterized and their functions are largely unknown. Here we report that ZNF23, a member of KRAB-ZFPs, inhibits cell cycle progression. ZNF23 protein localized to the nucleus and was ubiquitously expressed in all tested normal tissues. However, the expression levels of ZNF23 protein were lost or greatly reduced in human cancer. Ectopic expression of ZNF23 led to enhancement of p27(kip-1) expression, growth inhibition and cell cycle arrest in G(1) phase. Downregulation of p27(kip-1) by siRNA against p27(kip-1) reversed growth inhibition induced by ZNF23. Furthermore, the growth-inhibitory effect of ZNF23 was p53-independent. Deletion analysis revealed that the effect of ZNF23 did not rely on its KRAB domain, but on the C-terminal zinc fingers. Thus, we have identified a new member of KRAB-ZNF superfamily with growth inhibitory ability and its downregulation may contribute to carcinogenesis. PMID- 17137576 TI - Identification of calreticulin as a marker for phagocytosis of apoptotic cells in Drosophila. AB - Apoptotic cell phagocytosis is initiated through the specific interaction between markers for phagocytosis present at the surface of targets and their receptors of phagocytes. Although many molecules have been proposed to be phagocytosis markers and receptors in mammals, information as to the identity of those molecules is limited for invertebrate animals. Calreticulin, a molecular chaperone that functions in the lumen of the endoplasmic reticulum, was recently reported to be the second general marker, the membrane phospholipid phosphatidylserine being the first, for mammalian apoptotic cells to be recognized by phagocytes. We here asked whether or not calreticulin serves as a marker for phagocytosis in Drosophila. Phagocytosis of apoptotic S2 cells by Drosophila hemocyte-derived l(2)mbn cells, which we previously showed to occur independent of phosphatidylserine, was inhibited by the addition of anti-calreticulin antibody. This inhibition was observed when the target cells, but not phagocytes, were pre incubated with the antibody. In addition, RNA interference-mediated reduction of calreticulin expression in apoptotic S2 cells, but not in l(2)mbn cells, reduced the level of phagocytosis. An immunocytochemical analysis revealed that calreticulin is widely distributed at the surface of viable S2 cells. After the induction of apoptosis, cell surface calreticulin seemed to form aggregates, with no change in its amount. Furthermore, in embryos of a mutant Drosophila strain that expresses calreticulin at a reduced level, the level of phagocytosis of apoptotic cells was about a half of that observed in embryos of a wild-type strain. These results collectively indicate that calreticulin is the first molecule to be identified as a marker for phagocytosis of apoptotic cells by Drosophila phagocytes. PMID- 17137577 TI - Edinger-Westphal and pharmacologically stimulated accommodative refractive changes and lens and ciliary process movements in rhesus monkeys. AB - During accommodation, the refractive changes occur when the ciliary muscle contracts, releasing resting zonular tension and allowing the lens capsule to mold the lens into an accommodated form. This results in centripetal movement of the ciliary processes and lens edge. The goal of this study was to understand the relationship between accommodative refractive changes, ciliary process movements and lens edge movements during Edinger-Westphal (EW) and pharmacologically stimulated accommodation in adolescent rhesus monkeys. Experiments were performed on one eye each of three rhesus monkeys with permanent indwelling electrodes in the EW nucleus of the midbrain. EW stimulated accommodative refractive changes were measured with infrared photorefraction, and ciliary process and lens edge movements were measured with slit-lamp goniovideography on the temporal aspect of the eye. Images were recorded on the nasal aspect for one eye during EW stimulation. Image analysis was performed off-line at 30 Hz to determine refractive changes and ciliary body and lens edge movements during EW stimulated accommodation and after carbachol iontophoresis to determine drug induced accommodative movements. Maximum EW stimulated accommodation was 7.36+/-0.49 D and pharmacologically stimulated accommodation was 14.44+/-1.21 D. During EW stimulated accommodation, the ciliary processes and lens edge moved centripetally linearly by 0.030+/-0.001 mm/D and 0.027+/-0.001 mm/D, with a total movement of 0.219+/-0.034 mm and 0.189+/-0.023 mm, respectively. There was no significant nasal/temporal difference in ciliary process or lens edge movements. 30-40 min after pharmacologically stimulated accommodation, the ciliary processes moved centripetally a total of 0.411+/-0.048 mm, or 0.030+/-0.005 mm/D, and the lens edge moved centripetally 0.258+/-0.014 mm, or 0.019+/-0.003 mm/D. The peaks and valleys of the ciliary processes moved by similar amounts during both supramaximal EW and pharmacologically stimulated accommodation. In conclusion, this study shows, for the first time, that the ciliary processes and lens edge move centripetally, linearly with refraction during EW stimulated accommodation. During pharmacological stimulation, the ciliary processes move to a greater extent than the lens edge, confirming that in adolescent monkeys, lens movement limits the accommodative optical change in the eye. PMID- 17137578 TI - Poly (ADP-ribose) polymerase inhibitor 3-aminobenzamide rescues N-methyl-N nitrosourea-induced photoreceptor cell apoptosis in Sprague-Dawley rats through preservation of nuclear factor-kappaB activity. AB - The activation of poly (ADP-ribose) polymerase (PARP) plays a pivotal role in mediating N-methyl-N-nitrosourea (MNU)-induced photoreceptor cell apoptosis. We examined the retinoprotective effects of the PARP inhibitor 3-aminobenzamide (3 AB) against MNU-induced retinal damage in relation to dose and timing of prescription, and the involvement of the transcription factor nuclear factor (NF) kappaB. Female Sprague-Dawley rats were intraperitoneally injected with 60 mg/kg MNU at 50 days of age, and were then immediately given a subcutaneous injection of 0, 1, 5, 10, 30 or 50 mg/kg of 3-AB, or were injected with 50 mg/kg 3-AB 12h before, concurrently, or 4, 6 or 12h after MNU. Rats were killed 3 and 7 days after MNU, and MNU-treated and 3-AB-injected retinas were compared with MNU untreated control retinas or MNU-treated/3-AB-uninjected retinas. Apoptosis in photoreceptor cells was detected by performing formamide-induced DNA denaturation and staining with anti-single-stranded DNA antibody. Retinal morphologies were compared and evaluated morphometrically using the photoreceptor cell ratio and retinal damage ratio as indices to evaluate the efficacy of 3-AB. We examined expression of the phosphorylated form of NF-kappaB and IkappaBalpha (p-NF-kappaB and p-IkappaBalpha, respectively) in retinas of MNU-treated rats concurrently treated with or without 50mg/kg 3-AB, compared with MNU-untreated control retinas. 3-AB dose-dependently suppressed photoreceptor cell apoptosis: 50mg/kg 3 AB injected concurrently with MNU completely rescued photoreceptor cell damage; 30 mg/kg 3-AB significantly reduced photoreceptor cell damage; 10 mg/kg 3-AB tended to suppress photoreceptor cell damage; or=4h after MNU, it did not exert a retinoprotective effect. p-NF-kappaB levels of MNU-treated rat retinas were significantly lower than those of MNU-untreated control retinas, while 50 mg/kg 3 AB injected concurrently with MNU preserved the p-NF-kappaB levels; p IkappaBalpha levels tended to decrease after MNU injection, compared with untreated control retinas, but the difference was not significant. Thus, 3-AB dose-dependently suppressed MNU-induced retinal damage, and 50mg/kg 3-AB injected concurrently with MNU completely rescued photoreceptor cell apoptosis via preservation of NF-kappaB activity. PMID- 17137579 TI - ATP binding site in the plant ADP-glucose pyrophosphorylase large subunit. AB - The ATP binding region in the catalytically inactive large subunit (LS) of the potato tuber ADP-glucose pyrophosphorylase was identified and investigated. Mutations at the ATP binding significantly affected not only the apparent affinities for ATP and Glc-1-P, and catalytic rate but also in many instances, sensitivity to 3-phosphoglycerate. The catalytic rates of the LS mutant enzymes correlated most strongly with changes in the affinity toward ATP, a relationship substantiated by photoaffinity labeling studies with azido-ATP analog. These results indicate that the LS, although catalytically defective, interacts cooperatively with the catalytic small subunit in binding substrates and effectors and, in turn, influencing net catalysis. PMID- 17137580 TI - Sexual swellings in wild white-handed gibbon females (Hylobates lar) indicate the probability of ovulation. AB - Conspicuous sexual swellings in the females of some primate species have been a focus of scientific interest since Darwin first wrote about them in 1871. To understand these visual signals, research focused on exaggerated sexual swellings of Old World primates. However, some primate species develop much smaller sexual swellings and it is as yet unclear if these smaller swellings can serve similar functions as those proposed for exaggerated swellings, i.e. advertising fertility to attract mates. We studied the temporal patterns of sexual swellings, timing of ovulation and female reproductive status in wild white-handed gibbons (Hylobates lar) at Khao Yai National Park, Thailand, where this species has a variable social organization. We established fecal progestogen profiles in fifteen cycles of eight cycling females and, to detect swellings outside the menstrual cycle, five pregnant and six lactating females. In 80% of menstrual cycles, ovulation and maximum swelling phase (duration: O 9.3 days; 42.8% of cycle length), overlapped tightly. The probability of ovulation peaked on day 3 of the maximum swelling period. Nevertheless, the temporal relationship between maximum swelling and probability of ovulation varied from day -1 to day 13 of the swelling period and three times ovulations fell outside the maximum swelling phase. The different swellings phases occurred in similar proportions in cycling and pregnant, but not lactating females, which were rarely swollen. Despite their smaller size, gibbons' sexual swellings probably serve functions similar to those suggested for exaggerated swellings by the graded-signal hypothesis, which predicts that sexual swellings indicate the probability of ovulation, without allowing males to pinpoint its exact time. PMID- 17137581 TI - Differential functions of tissue factor in the trans-activation of cellular signalling pathways. AB - In this study we examined the ability of tissue factor (TF) alone, or in conjunction with factor VIIa, factor Xa and TFPI in activating a number of key signalling pathways associated with cellular growth, stress and differentiation responses in human endothelial cells. We used luciferase reporter systems to demonstrate the activation of p42/44 MAPK by the TF-FVIIa complex, mediated via the PAR1 receptor. TF alone was capable of interacting with the cell surface and was sufficient to activate the JNK-SAPK pathway and subsequently AP-1, but the level of activation was enhanced by the activity of FXa on PAR1 and 2. Furthermore, the phosphorylated form of the transmembrane-cytoplasmic domain of TF was directly responsible for activation of these pathways. CREB activation occurred in response to TF-FVIIa in a non-protease dependent manner but was lowered on addition of FXa. Finally, NFkappaB activation occurred in response to FVIIa or FXa, with the latter exhibiting higher levels of activation. In conclusion, we have shown that TF is capable of activating differing signalling pathways, via more than one mechanism. The differential influence of TF is modified depending on the presence of other coagulation factors and ultimately acts as a deciding factor in the determination of cellular fate. PMID- 17137582 TI - Moderately elevated plant sterol levels are associated with reduced cardiovascular risk--the LASA study. AB - Functional foods with supplementation of plant sterols are already used by millions of people. However, at the same time it is current scientific thinking that elevation of plant sterols in the circulation causes coronary heart disease. Therefore, this study aimed to define the risk for coronary heart disease associated with moderately high plant sterol plasma levels in a cohort of elderly. In this study, we evaluated the association between plant sterols and coronary heart disease in a cohort of 1242 subjects older than 65 years, participating at the Longitudinal Aging Study Amsterdam (LASA). Concentrations of sitosterol, campesterol, brassicasterol and stigmasterol were assessed using highly sensitive and specific gas chromatography-mass spectrometry-selected ion monitoring. Plant sterol concentrations (and their ratios to cholesterol) were slightly, however, significantly lower in patients with coronary heart disease. Moreover, high plasma concentrations of a marker plant sterol, sitosterol, were associated with a markedly reduced risk for coronary heart disease (OR 0.78, CI 0.62-0.98, p<0.05). In contrast neither plant stanols (sitostanol or campestanol) nor the cholesterol synthesis markers (lathosterol, lanosterol and desmosterol) nor their ratios to cholesterol were significantly different in the study groups. These data suggest that plant sterols could have neutral or even protective effects on development of coronary heart disease, which have to be confirmed in interventional trials. PMID- 17137583 TI - Slow heart rate recovery after exercise is associated with carotid atherosclerosis. AB - OBJECTIVE: Slow heart rate recovery (HRR) after exercise is an estimate of impaired parasympathetic tone and predictor of all-cause and cardiovascular mortality. Carotid atherosclerosis is associated with high risk of developing coronary heart disease (CHD) and stroke. We tested the hypothesis that slow HRR is associated with carotid atherosclerosis in a cross-sectional study of 12,712 middle-aged men (age 49.1+/-8.9 years). METHODS: Carotid atherosclerosis was measured using B-mode ultrasonography and defined as stenosis >25% and/or intima media thickness >1.2mm. HRR was calculated as the difference between peak heart rate during a graded exercise treadmill test and heart rate 2 min after cessation of exercise. RESULTS: The prevalence of carotid atherosclerosis was 8.4%. The prevalence of atherosclerosis was significantly higher among subjects in the lowest (<44 bpm) versus the highest (>61 bpm) quartile of HRR (14.4% versus 4.1%, p<0.001). In multivariable logistic regression models adjusted for established CHD risk factors, inflammatory markers, and exercise capacity, subjects in the lowest quartile of HRR (<44 bpm) were 1.50 times (95% CI: 1.13-2.00) more likely to have carotid atherosclerosis than subjects in the highest quartile (HRR>61 bpm). CONCLUSIONS: Slow heart rate recovery after exercise, an index of decreased parasympathetic activity, is associated with carotid atherosclerosis independent of established risk factors in middle-age men. PMID- 17137584 TI - Investigating full-field deformation of planar soft tissue under simple-shear tests. AB - Finite simple shear test characteristics like specimen geometry and boundary conditions could affect the deformation homogeneity during the test. In order to ensure that the parameters of constitutive equations obtained from finite simple shear tests are appropriate, the deformation homogeneity of the specimen during simple-shear test should be examined. The Fourier transform moire method (FTM) was used to examine the deformation uniformity of a porcine skin specimen in a finite simple shear test. The effects of clamping prestrain (0.15 and 0.3 engineering strain) and specimen geometry (5x5, 5x3.75, and 5x2.5cm) were investigated. These effects include in-plane deformation altered by clamping prestrain, slippage between specimen and clamps, and out-of-plane deformation. The experimental results showed that the wide specimen had more severe deformation alteration by clamping prestrain and was easier to slip out of the clamps when the shear angle is large. Furthermore, in all test configurations, the out-of-plane deformation is significant when the shear angle is large, and a narrow specimen is prone to have out-of-plane deformation. This study may provide guidelines for the selection of specimen aspect ratio and clamping prestrain when studying the material response of soft tissues under simple-shear tests. PMID- 17137585 TI - Solid phase extraction, multidimensional gas chromatography mass spectrometry determination of four novel aroma powerful ethyl esters. Assessment of their occurrence and importance in wine and other alcoholic beverages. AB - A method for the quantitative determination of four powerful aromatic ethyl esters recently identified in some wines has been developed, validated and applied to the determination of these compounds in different samples of wine, whisky and brandy. Ethyl 2-, 3-, and 4-methylpentanoate and ethyl cyclohexanoate are extracted from 100ml of sample by solid phase extraction (SPE) on a 200mg LiChrolut EN bed. Major compounds are eliminated by rinsing with a water-methanol (50:50) solution containing 1% sodium bicarbonate, and analytes are eluted with 1.5ml of dichloromethane. Fifty microlitres of this extract are then injected in a multidimensional gas chromatography-mass spectromety (GC-GC-MS) system. Recoveries in the SPE are quantitative. Method repeatability is satisfactory (5 12% for a 5-10ngl(-1) level, and less than 7% for 25-50ngl(-1) level), the method linearity holds along the whole range of occurrence of analytes (2-2700ngl(-1)), and the signal is independent on the matrix. Method detection limits are below 1ngl(-1) in all cases. Results suggest that these compounds are formed by the slow esterification with ethanol of the corresponding acids formed by different microorganisms. The levels of these compounds are above the corresponding thresholds in most samples of aged wines or distillates, but are particularly high in some sweet wines, whiskeys and brandies where they may constitute the most important contributors to the sweet-fruity notes reaching concentrations up to 85-350 times higher than the corresponding odor thresholds. PMID- 17137586 TI - Determination of sulfonamide compounds in sewage and river by mixed hemimicelles solid-phase extraction prior to liquid chromatography-spectrophotometry. AB - Mixed hemimicelles-based solid-phase extraction was investigated for the preconcentration of five sulfonamides from environmental water samples prior to HPLC-spectrophotometry determination in this paper. A comparative study on the use of sodium dodecyl sulfate (SDS) coating gamma-alumina or octadecyltrimethylammonium bromide (OTMABr) and OTMABr coating silica as sorbent materials were presented. The five analytes (sulfadiazine (SDA), sulfathiazole (STA), sulfapyridine (SPD), sulfamethazine (SMZ) and sulfamethoxazole (SMX)) were quantitatively adsorbed on OTMABr-gamma-alumina and OTMABr-silica mixed hemimicelles, but OTMABr-gamma-alumina was not adopted because it worked at a high pH (around 10), instead, OTMABr-silica was selected to overcoming the pH restriction. The analytes retained on the cartridge were quantitatively desorbed with suitable amounts of methanol. Factors influencing the extraction efficiency, such as the amount of surfactant, pH of sample and breakthrough volume were discussed. The proposed method had been applied to determining the five sulfonamides in several environmental water samples and concentration factors of 300 and 600 for SDA and other four analytes were achieved, respectively. Detection limits obtained ranged between 0.15 and 0.35microg/L for this five sulfonamides under the optimized conditions. The accuracy of the method was evaluated by recovery measurements on spiked samples, and good recovery results (89-113%) with precision of 3-6% were achieved. PMID- 17137587 TI - Monolithic silica columns of various format in automated sample clean up/multidimensional liquid chromatography/mass spectrometry for peptidomics. AB - The following particulate and monolithic silica columns were implemented in a fully automated and flexible multidimensional LC/MS system with integrated sample clean-up, to perform the analysis of endogeneous peptides from filtered urine and plasma samples: restricted access sulphonic acid strong cation-exchanger (RAM SCX) for sample clean-up, RP 18 Chromolith guard columns as trap columns and 100 microm I.D. monolithic RP 18 fused silica capillary columns as last LC dimension. The results show sufficient overall system reproducibility and repeatability. Implementation of monolithic silica columns added an additional flexibility with respect to flow rate variation and adjustment due to the low column back pressures. Also, monolithic columns showed a lower clogging rate in long-term usage for biological samples as compared to particulate columns. The applied system set-up was tested to be useful for the routine peptide screening in search of disease biomarkers. PMID- 17137588 TI - Novel Janus Cu2(OH)2CO3/CuS microspheres prepared via a Pickering emulsion route. AB - Janus Cu2(OH)2CO3/CuS microspheres were prepared via a Pickering emulsion route for the first time. By treating the Janus Cu2(OH)2CO3/CuS microspheres with dilute hydrochloric acid, ringent Cu2(OH)2CO3/CuS core/shell microspheres and ringent CuS shells were obtained. The hatch size of the ringent CuS shells increased with the increase of the hydrophobicity of the precursor Cu2(OH)2CO3 microspheres. Scanning electron microscopy, X-ray diffraction, energy dispersion spectra, and particle size analysis were used to characterize the products thus formed. PMID- 17137589 TI - Efficient expression of recombinant human monoclonal antibodies in Drosophila S2 cells. AB - We have explored the Drosophila S2 cell line for expression of Ig molecules isolated as Fab or scFv cDNA from phage-displayed libraries. We present a series of vectors for inducible expression and secretion of human Ig heavy (HC) and light chains (LC), both on separate plasmids and in combination constructs. Both HC (tested as human gamma(1)) and LC (human kappa) could be expressed separately and were secreted into the medium, confirming previous reports. When the combination vector carrying both the HC and LC cDNA, as well as when the HC and LC vectors were co-transfected, complete IgG1 was found in the medium. Transient transfection resulted in production levels of 0.5-1 mg/l. Stable cell lines could be established within 2-3 weeks. After 10-12 days of expression from such cell lines, Ig molecules accumulated and the medium contained typically 5-35 mg/l of IgG1. The IgG in these preparations was purified to more than 90% purity on protein G columns. Binding characteristics for IgG of the same clone expressed in S2 cells or mammalian cells were indistinguishable. The main advantages with this system compared to mammalian expression were its robustness and the much faster establishment of stable, high level producing cell lines. PMID- 17137590 TI - A novel disulfide bond in the SH2 Domain of the C-terminal Src kinase controls catalytic activity. AB - The SH2 domain of the C-terminal Src kinase [Csk] contains a unique disulfide bond that is not present in other known SH2 domains. To investigate whether this unusual disulfide bond serves a novel function, the effects of disulfide bond formation on catalytic activity of the full-length protein and on the structure of the SH2 domain were investigated. The kinase activity of full-length Csk decreases by an order of magnitude upon formation of the disulfide bond in the distal SH2 domain. NMR spectra of the fully oxidized and fully reduced SH2 domains exhibit similar chemical shift patterns and are indicative of similar, well-defined tertiary structures. The solvent-accessible disulfide bond in the isolated SH2 domain is highly stable and far from the small lobe of the kinase domain. However, reduction of this bond results in chemical shift changes of resonances that map to a cluster of residues that extend from the disulfide bond across the molecule to a surface that is in direct contact with the small lobe of the kinase domain in the intact molecule. Normal mode analyses and molecular dynamics calculations suggest that disulfide bond formation has large effects on residues within the kinase domain, most notably within the active-site cleft. Overall, the data indicate that reversible cross-linking of two cysteine residues in the SH2 domain greatly impacts catalytic function and interdomain communication in Csk. PMID- 17137591 TI - Crystallographic and in silico analysis of the sialoside-binding characteristics of the Siglec sialoadhesin. AB - The Siglec family of receptors mediates cell-surface interactions through recognition of sialylated glycoconjugates. Previously reported structures of the N-terminal domain of the Siglec sialoadhesin (SnD1) in complex with various sialic acid analogs revealed the structural template for sialic acid binding. To characterize further the carbohydrate-binding properties, we have determined the crystal structures of SnD1 in the absence of ligand, and in complex with 2-benzyl Neu5NPro and 2-benzyl-Neu5NAc. These structures reveal that SnD1 undergoes very few structural changes on ligand binding and detail how two novel classes of sialic acid analogs bind, one of which unexpectedly can induce Siglec dimerization. In conjunction with in silico analysis, this set of structures informs us about the design of putative ligands with enhanced binding affinities and specificities to different Siglecs, and provides data with which to test the effectiveness of different computational drug design protocols. PMID- 17137592 TI - The transition state of the ras binding domain of Raf is structurally polarized based on Phi-values but is energetically diffuse. AB - The ras binding domain (RBD) of the Ser/Thr kinase c-Raf/Raf-1 spans 78 residues and adopts a structure characteristic of the beta-grasp ubiquitin-like topology. Recently, the primary sequence of Raf RBD has been nearly exhaustively mutated experimentally by insertion of stretches of degenerate codons, which revealed sequence conservation and hydrophobic core organization similar to that found in an alignment of beta-grasp ubiquitin-like proteins. These results now allow us to examine the relationship between sequence conservation and the folding process, particularly viewed through the analysis of transition state (TS) structure. Specifically, we present herein a protein engineering study combining classic truncation (Ala/Gly) and atypical mutants to predict folding TS ensemble properties. Based on classical Phi-value analysis, Raf RBD TS structure is particularly polarized around the N-terminal beta-hairpin. However, all residues constituting the inner layer of the hydrophobic core are involved in TS stabilization, although they are clearly found in a less native-like environment. The TS structure can also be probed by a direct measure of its destabilization upon mutation, DeltaDeltaG(U-++). Viewed through this analysis, Raf RBD TS is a more diffuse structure, in which all residues of the hydrophobic core including beta-strands 1, 2, 3 and 5 and the major alpha-helix play similar roles in TS stabilization. In addition, Phi-values and DeltaDeltaG(U-++) reveal striking similarities in the TS of Raf RBD and ubiquitin, a structural analogue displaying insignificant sequence identity (<12%). However, ubiquitin TS appears more denatured-like and polarized around the N-terminal beta-hairpin. We suggest that analysis of Phi-values should also consider the direct impact of mutations on differences in free energy between the unfolded and TS (DeltaDeltaG(U-++)) to ensure that the description of TS properties is accurate. Finally, the impact of these findings on the modeling of protein folding is discussed. PMID- 17137593 TI - Crystal structures of human IPP isomerase: new insights into the catalytic mechanism. AB - Type I isopentenyl diphosphate (IPP): dimethylally diphosphate (DMAPP) isomerase is an essential enzyme in human isoprenoid biosynthetic pathway. It catalyzes isomerization of the carbon-carbon double bonds in IPP and DMAPP, which are the basic building blocks for the subsequent biosynthesis. We have determined two crystal structures of human IPP isomerase I (hIPPI) under different crystallization conditions. High similarity between structures of human and Escherichia coli IPP isomerases proves the conserved catalytic mechanism. Unexpectedly, one of the hIPPI structures contains a natural substrate analog ethanol amine pyrophosphate (EAPP). Based on this structure, a water molecule is proposed to be the direct proton donor for IPP and different conformations of IPP and DMAPP bound in the enzyme are also proposed. In addition, structures of human IPPI show a flexible N-terminal alpha-helix covering the active pocket and blocking the entrance, which is absent in E. coli IPPI. Besides, the active site conformation is not the same in the two hIPPI structures. Such difference leads to a hypothesis that substrate binding induces conformational change in the active site. The inhibition mechanism of high Mn(2+) concentrations is also discussed. PMID- 17137594 TI - Identification of the LEDGF/p75 binding site in HIV-1 integrase. AB - Lens epithelium-derived growth factor (LEDGF)/p75 is an important cellular co factor for human immunodeficiency virus (HIV) replication. We originally identified LEDGF/p75 as a binding partner of integrase (IN) in human cells. The interaction has been mapped to the integrase-binding domain (IBD) of LEDGF/p75 located in the C-terminal part. We have subsequently shown that IN carrying the Q168A mutation remains enzymatically active but is impaired for interaction with LEDGF/p75. To map the integrase/LEDGF interface in more detail, we have now identified and characterized two regions within the enzyme involved in the interaction with LEDGF/p75. The first region centers around residues W131 and W132 while the second extends from I161 up to E170. For the different IN mutants the interaction with LEDGF/p75 and the enzymatic activities were determined. IN(W131A), IN(I161A), IN(R166A), IN(Q168A) and IN(E170A) are impaired for interaction with LEDGF/p75, but retain 3' processing and strand transfer activities. Due to impaired integration, an HIV-1 strain containing the W131A mutation in IN displays reduced replication capacity, whereas virus carrying IN(Q168A) is replication defective. Comparison of the wild-type IN-LEDGF/p75 co crystal structure with that of the modelled structure of the IN(Q168A) and IN(W131A) mutant integrases corroborated our experimental data. PMID- 17137595 TI - Crystal structures of a poxviral glutaredoxin in the oxidized and reduced states show redox-correlated structural changes. AB - Glutaredoxins act as reducing agents for the large subunit of ribonucleotide reductase (R1) in many prokaryotes and eukaryotes, including humans. The same relationship has been proposed for the glutaredoxin and R1 proteins expressed by all orthopoxviruses, including vaccinia, variola, and ectromelia virus. Interestingly, the orthopoxviral proteins share 45% and 78% sequence identity with human glutaredoxin-1 (Grx-1) and R1, respectively. To study structure function relationships of the vertebrate Grx-1 family, and reveal potential viral adaptations, we have determined crystal structures of the ectromelia virus glutaredoxin, EVM053, in the oxidized and reduced states. The structures show a large redox-induced conformational rearrangement of Tyr21 and Thr22 near the active site. We predict that the movement of Tyr21 is a viral-specific adaptation that increases the redox potential by stabilizing the reduced state. The conformational switch of Thr22 appears to be shared by vertebrate Grx-1 and may affect the strictly conserved Lys20. A crystal packing-induced structural change in residues 68-70 affects the GSH-binding loop, and our structures reveal a potential interaction network that connects the GSH-binding loop and the active site. EVM053 also exhibits a novel cis-proline (Pro53) in a loop that has been shown to contribute to R1-binding in Escherichia coli Grx-1. The cis-peptide bond of Pro53 may be required to promote electrostatic interactions between Lys52 and the C-terminal carboxylate of R1. Finally, dimethylarsenite was covalently attached to Cys23 in one reduced EVM053 structure and our preliminary data show that EVM053 has dimethylarsenate reductase activity. PMID- 17137596 TI - Failure to detect an association between the serotonin transporter gene and borderline personality disorder. PMID- 17137597 TI - Computer-based training for safety: comparing methods with older and younger workers. AB - INTRODUCTION: Computer-based safety training is becoming more common and is being delivered to an increasingly aging workforce. Aging results in a number of changes that make it more difficult to learn from certain types of computer-based training. Instructional designs derived from cognitive learning theories may overcome some of these difficulties. METHODS: Three versions of computer-based respiratory safety training were shown to older and younger workers who then took a high and a low level learning test. RESULTS: Younger workers did better overall. Both older and younger workers did best with the version containing text with pictures and audio narration. DISCUSSION: Computer-based training with pictures and audio narration may be beneficial for workers over 45 years of age. IMPACT ON INDUSTRY: Computer-based safety training has advantages but workers of different ages may benefit differently. Computer-based safety programs should be designed and selected based on their ability to effectively train older as well as younger learners. PMID- 17137598 TI - The estimated reduction in the odds of loss-of-control type crashes for sport utility vehicles equipped with electronic stability control. AB - INTRODUCTION: Using data from the NASS General Estimates System (GES), the method of induced exposure was used to assess the effects of electronic stability control (ESC) on loss-of-control type crashes for sport utility vehicles. METHOD: Sport utility vehicles were classified into crash types generally associated with loss of control and crash types most likely not associated with loss of control. Vehicles were then compared as to whether ESC technology was present or absent in the vehicles. A generalized additive model was fit to assess the effects of ESC, driver age, and driver gender on the odds of loss of control. In addition, the effects of ESC on roads that were not dry were compared to effects on roads that were dry. RESULTS: Overall, the estimated percentage reduction in the odds of a loss-of-control crash for sport utility vehicles equipped with ESC was 70.3%. Both genders and all age groups showed reduced odds of loss-of-control crashes, but there was no significant difference between males and females. With respect to driver age, the maximum percentage reduction of 73.6% occurred at age 27. The positive effects of ESC on roads that were not dry were significantly greater than on roads that were dry. PMID- 17137599 TI - Creep behavior of commonly used suture materials in abdominal wall surgery. AB - BACKGROUND: The incidence of incisional hernia after abdominal wall closure is high. Furthermore, recurrence is a significant complication after correction of all abdominal wall hernias. Besides surgeon- and patient-related factors, in this experimental study a third factor, i.e., creep behavior of suture materials, is introduced and evaluated. MATERIALS AND METHODS: Creep measurements were performed on 0 and 2-0 Prolene (Ethicon, Johnson & Johnson Intl., Somerville, NJ) and 1 and 2-0 PDSII (Ethicon, Johnson & Johnson Intl.) sutures. Two different loads were used representing normal intra-abdominal pressure (IAP) and pathological IAP. A mean percentage of elongation was calculated for each type of suture material. Statistical analysis was performed using analysis of variance. RESULTS: All suture materials showed significant (3-51%) creep behavior. Prolene sutures showed more creep than PDSII sutures in both loading conditions. CONCLUSIONS: As significant creep was demonstrated for commonly used suture materials, creep might be a significant influential factor with regard to the etiology of incisional hernias and recurrence after abdominal wall hernia repair. PMID- 17137600 TI - Isolated facial myorhythmia. AB - Myorhythmia, characterized by relatively rhythmic and regular 1- to 3-Hz muscle contractions, may involve isolated limbs or in combination with other various body regions. Isolated facial myorhythmia is rare. We highlight the clinical and electrophysiologic features of a unique case of isolated facial myorhythmia. Extensive investigations did not reveal any conclusive secondary causes. Dopaminergic dysfunction could be one of the etio-pathologic factors. However, the pathological basis why the facial region was selectively involved in our patient is unclear. Early recognition of the symptoms and signs would facilitate investigations for an underlying cause. PMID- 17137601 TI - Nonprogressive juvenile-onset spinal muscular atrophy: A clinico-radiological and CAG repeat study of androgen receptor gene. AB - BACKGROUND: Occurrence of nonprogressive juvenile-onset spinal muscular atrophy (SMA) predominantly in males suggests a possibility of X-linked disorder but there is no such report addressing this problem. AIMS: To evaluate CAG repeat expansion of androgen receptor (AR) gene in patients with nonprogressive juvenile onset SMA. SETTING: Tertiary medical teaching institute. SUBJECTS AND METHODS: Patients fulfilling the diagnostic criteria of nonprogressive juvenile-onset SMA were included. Detailed clinical evaluation and pedigree charting were done in all. Nerve conduction study, electromyography and cervical spinal MRI were carried out. From peripheral venous blood, DNA was separated and AR gene CAG repeat exon polymorphism was assayed using polymerase chain reaction (PCR) in conjugation with genotyping and Gene scan soft ware. Number of CAG repeats was compared with normal controls. RESULTS: 25 patients with nonprogressive juvenile onset SMA from 24 families were included and their mean age was 22.2 years. Age at the time of disease onset ranged between 15 and 30 years with a mean duration of illness 2.6 years. None of the patients had testicular atrophy or gynecomastia. C7-T1 myotomal wasting and weakness although was unilateral to begin with but became bilateral in 16 and 4 more patients had evidences of subclinical involvement of the other side as revealed by EMG. Spinal MRI revealed cord atrophy at C6-8 vertebral level in 16 patients. CAG repeat study of AR gene was carried out in 16 patients. The number of CAG repeats in patients ranged between 15 and 39 (median 21) which were within the normal range. CONCLUSION: Abnormal CAG repeat expansion of AR gene is not found in patients with nonprogressive juvenile-onset SMA. PMID- 17137602 TI - Scale invariant correlations between genes and SNPs on Human chromosome 1 reveal potential evolutionary mechanisms. AB - The local density of gene structures and single nucleotide polymorphisms (SNPs) along human chromosomes appears inhomogeneous. In chromosome 1, the density patterns from both these elements are shown here to exhibit similar scale invariant clustering, as well as long-ranged and scale invariant auto- and cross correlations. The local densities of these elements sites can be accurately represented by the scale invariant exponential dispersion models, a group of stochastic models that act as limiting distributions for a wide range of generalized linear models. The scale invariant Poisson-gamma (PG) distribution is the most applicable of these models, since it describes the above findings and it lends itself to a stochastic mechanism for the accumulation of segmental chromosomal changes. This PG model describes the summation of neutral chromosomal mutations, deletions, rearrangements and recombinations, within chromosomal segments that are distinguished by their evolutionary genealogies. Scale invariance is a necessary property if such a description is to remain valid at different measurement scales. The observed density patterns, and proposed model, presumably represent the convergent summation of multiple stochastic processes within the evolutionary history of the chromosome. PMID- 17137603 TI - Buthionine sulfoximine causes endothelium dependent hyper-relaxation and hypoadiponectinemia. AB - A close relationship between oxidative stress, endothelial dysfunction, and hypoadiponectinemia has been observed. The present study was performed to investigate how glutathione depletion via buthionine sulfoximine (BSO) administration affects endothelial function and adiponectin levels in rats. Acetylcholine (Ach)-induced vasodilation was significantly enhanced in BSO treated rats, compared with control rats. This was completely abolished by L NAME, and Ach-induced vasodilation was not observed in the aorta without endothelium. These results suggest that Ach-induced hyper-relaxation of the aorta in BSO-treated rats is completely dependent on the presence of endothelium and mediated by changes in eNOS activity. Catalase significantly inhibited this relaxation to Ach and no effect of catalase on sodium nitroprusside-induced relaxation of the aorta without endothelium was observed in BSO-treated rats. Thus, hyper-relaxation of the aorta in BSO-treated rats is likely caused by H2O2 in addition to NO produced by the endothelium via an eNOS-dependent mechanism. Hypoadiponectinemia and decreased levels of adiponectin mRNA in adipose tissue were observed in BSO-treated rats. Protein expression of eNOS and SODs (SOD-1 and SOD-2) in the aorta was increased and plasma NOx levels were decreased in BSO treated rats. Our results suggest that oxidative stress induced by BSO causes eNOS uncoupling and hyper-relaxation by producing H2O2, and that BSO-induced oxidative stress causes hypoadiponectinemia, probably by increasing H2O2 production in adipose tissue. PMID- 17137604 TI - Signaling pathways involved in pilocarpine-induced mucin secretion in rat submandibular glands. AB - We have studied the signaling pathways involved in pilocarpine-induced mucin release in rat submandibular slices. Pilocarpine produced a significant increment of PGE2 levels and a positive (r=0.8870) and significant (p=0.0077) correlation between PGE2 production and mucin released was determined. The participation of PGE2 was confirmed by the use of indomethacin (indo) and of acetyl salicylic acid (ASA), cyclooxygenase inhibitors, which inhibited pilocarpine-induced mucin release. The muscarinic receptors involved in the regulation of mucin release were identified as M1 and M4 by the use of the selective acetylcholine receptors (mAChR) antagonists, pirenzepine, AF-DX 116, 4-DAMP and tropicamide. The secretory process was dependent on both, intracellular and extracellular calcium pools since it was inhibited by thapsigargin and verapamil. Cyclic AMP, nitric oxide synthase and PKC also participated in pilocarpine-induced mucin release. It is concluded that pilocarpine, by activation the M1 and M4 mAChR subtypes induces an increase of intracellular Ca2+ concentration ([Ca2+]I) and elevates cAMP levels, which in turn stimulates COX, PKC and NOS and promotes mucin exocytosis. PGE2 released induces cAMP accumulation which, together with PKC are involved in the PGE2 increased Ca2+/cAMP-regulated exocytosis. Thus, cAMP accumulation induced by cholinergic stimulation is, in part, the result of PGE2 production. PMID- 17137605 TI - Microarray analysis of gene expression profile in the corpus cavernosum of hypercholesterolemic rats after chronic treatment with PDE5 inhibitor. AB - Gene expression changes in the corpus cavernosum of hypercholesterolemic rats were not fully assessed, which were not previously known to be associated with hypercholesterolemia-related erectile dysfunction (ED). To provide molecular insight into pathophysiology of hypercholesterolemia-related ED and to investigate the effects of Udenafil, a phosphodiesterase type 5 (PDE5) inhibitor, on gene expression, we performed microarray gene expression analysis via gene discovery methods using GenoCheck platinum cDNA chip (Ansan, S. Korea). Sixteen male Sprague-Dawley rats were fed 2% cholesterol diet for 5 months. Half of them were orally treated with Udenafil (20 mg/kg/day) simultaneously. Eight age matched rats fed normal diet were served as normal control. RNA was extracted from corpus cavernosum and microarray analysis was performed. Decreased erectile responses and hypercholesterolemia were observed in hypercholesterolemic control group. In microarray analysis, 122 candidate genes were noted to be altered based on the magnitude of expression changes, which includes 44 down-regulated and 78 up-regulated genes compared with the age-matched normal controls. These changes were, however, significantly attenuated by treatment with Udenafil. Out of the 78 up-regulated genes, 8 genes were significantly decreased by the chronic treatment with Udenafil. The altered genes were cytochrome oxidase biogenesis protein OXA1, skeletal muscle myosin heavy chain, lipophilin, fast skeletal muscle isoforms beta/alpha, myosin light chain 3, cytochrome c oxidase, adipocyte fatty acid binding protein and one EST gene. In contrast, among the 44 down-regulated genes, Kruppel-like factor 5 and cyclin D1 genes were increased after the Udenafil treatment. These results provide the molecular basis for understanding the pathogenesis of hypercholesterolemia-related ED and offer clues on determining the underlying action mechanism of a PDE5 inhibitor. PMID- 17137606 TI - A new type of orally active anti-diabetic Zn(II)-dithiocarbamate complex. AB - In order to find orally active Zn(II) complexes that can treat diabetes mellitus (DM) at low doses, four new Zn(II)-dithiocarbamate complexes with Zn(II)-sulfur coordination bonds were prepared and their in vitro insulinomimetic activity and in vivo anti-diabetic ability were evaluated. Among the Zn(II)-dithiocarbamate complexes, the bis(pyrrolidine-N-dithiocarbamate)zinc(II) (Zn(pdc)(2)) complex was found to be the most effective in terms of inhibiting free fatty acid-release and enhancing glucose-uptake in adipocytes. After oral administration of the Zn(pdc)(2) complex to KK-A(y) mice with obesity and type 2 DM, we observed that the high blood glucose levels in the mice were lowered from approximately 500 mg/dL to 350 mg/dL within 6 days, and the effect was maintained during the administration period. Also, indicators of insulin resistance such as serum insulin, leptin, and triglyceride levels were also reduced compared with those in untreated mice. Moreover, the Zn(pdc)(2) complex improved not only the hypertension in the mice, but also the adiponectin level in the serum. On the basis of the results, the Zn(pdc)(2) complex is proposed to improve hyperglycemia and insulin resistance in type 2 DM animals on daily oral administrations. PMID- 17137607 TI - Function and regulation of taurine transport at the inner blood-retinal barrier. AB - In the retina, taurine exerts a number of neuroprotective functions as an osmolyte and antioxidant. The purpose of the present study was to elucidate the taurine transport system(s) at the inner blood-retinal barrier (BRB). [(3)H]Taurine transport at the inner BRB was characterized using in vivo integration plot analysis and a conditionally immortalized rat retinal capillary endothelial cell line (TR-iBRB2 cells). The expression of the taurine transporter (TauT) was demonstrated by RT-PCR and immunoblot analyses. The apparent influx permeability clearance of [(3)H]taurine in the rat retina was found to be 259 muL/(ming retina), supporting carrier-mediated influx transport of taurine at the BRB. [(3)H]Taurine uptake by TR-iBRB2 cells was Na(+)-, Cl(-)- and concentration dependent with a K(m) of 22.2 muM and inhibited by TauT inhibitors, such as beta alanine and hypotaurine. RT-PCR and immunoblot analyses demonstrated that TauT is expressed in TR-iBRB2 and primary cultured human retinal endothelial cells. The uptake of [(3)H]taurine and the expression of TauT mRNA in TR-iBRB2 cells increased under hypertonic conditions but decreased following pretreatment with excess taurine. In conclusion, TauT most likely mediates taurine transport and regulate taurine transport at the inner BRB. PMID- 17137608 TI - Comparison of retinal arteriolar and venular variability in healthy subjects. AB - Study of retinal autoregulation is important because vascular dysfunction is a precursor of many retinal diseases. Previous research has focused on venular blood flow because the minimal venular pulsatility was thought to provide more reproducible results. This study compared the variability of arteriolar and venular blood flow measurements in response to isocapnic hyperoxia, a provocation known to constrict blood vessels and reduce blood velocity. Data was collected using a non-invasive laser Doppler instrument that permitted the simultaneous measurement of retinal blood velocity and vessel diameter, allowing the derivation of blood flow. Measurements were collected from 20 young subjects before, during and after exposure to hyperoxia. Isocapnia was maintained throughout hyperoxia using a previously validated sequential re-breathing circuit. Arteriolar and venular diameters decreased during hyperoxia by 8.7% (p=0.0001) and 14.2% (p=0.0001), respectively. Hyperoxia caused significant decreases in arteriolar and venular blood velocity (31.2%, p=0.0001 and 18.0%, p=0.0001, respectively) and flow (43.2%, p=0.0001 and 40.0%, p=0.0002, respectively). The coefficients of variation for intra-individual measurements of diameter, velocity and flow were comparable in magnitude between the two vessel types. Measures of arteriolar pulsatility, such as Pulsatility ratio, Resistivity ratio and Pulsatility index, increased significantly during hyperoxia, indicating increased downstream vascular resistance. We conclude that retinal arterioles and venules provide equally reproducible results for autoregulation studies and that arteriolar pulsatility profiles provide additional useful information regarding vascular resistance. PMID- 17137609 TI - Modulation of food reward by adiposity signals. AB - Extensive historical evidence from the drug abuse literature has provided support for the concept that there is functional communication between central nervous system (CNS) circuitries which subserve reward/motivation, and the regulation of energy homeostasis. This concept is substantiated by recent studies that map anatomical pathways, or which demonstrate that hormones and neurotransmitters associated with energy homeostasis regulation can directly modulate reward and motivation behaviors. Studies from our laboratory have focused specifically on the candidate adiposity hormones, insulin and leptin, and show that these hormones can decrease performance in behavioral paradigms that assess the rewarding or motivating properties of food. Additionally we and others have provided evidence that the ventral tegmental area may be one direct target for these effects, and we are currently exploring other potential anatomical targets. Finally, we are beginning to explore the interaction between adiposity signals, chronic maintenance diet of rats, and different types of food rewards to more closely simulate the current food environments of Westernized societies including the U.S. We propose that future studies of food reward should include a more complex environment in the experimental design that takes into account abundance and variety of rewarding foods, psychological stressors, and choices of reward modalities. PMID- 17137610 TI - "Chromoplast" development in arbuscular mycorrhizal roots. AB - The accumulation of apocarotenoids in arbuscular mycorrhizal (AM) roots suggests a dramatic reorganization of the plastids responsible for the biosynthesis of these compounds. This review describes the cytological and biochemical characterization of this phenomenon. The results presented suggest that plastids are key organelles for the establishment of the symbiotic interface of the AM symbiosis. In addition, a complex interplay of various plant cell components during the different functional phases of this interface is suggested. Arbuscule degradation appears to be of particular interest, as it correlates with the formation of the most extensive plastid structures and with apocarotenoid accumulation. PMID- 17137611 TI - Simplification of the IAST for activated carbon adsorption of trace organic compounds from natural water. AB - Recent studies have shown that the ideal adsorbed solution theory (IAST) coupled with the concept of equivalent background compound (EBC) can be simplified for describing trace organic compound adsorption from natural water, provided that the adsorbent surface loading is dominated by competing natural organic matter. The resulting simplified IAST has been used to reduce the complexity of kinetic models for various dynamic adsorption processes. In order to be correctly applied, however, the simplified IAST requires some additional clarification and a quantitative evaluation of the deviation caused by the simplifying assumption. In this study, we derive a simple equation that relates the relative deviation of the simplified IAST directly to the molar ratio of EBC and trace organic compound surface loadings and their Freundlich isotherm exponents. We then verify the simplified IAST using the original IAST and experimental isotherm data from the literature for trace organic compounds at various initial concentrations in natural water. By further assuming that the adsorbed amount of the EBC is substantially greater than what remains in solution, a new pseudo single-solute isotherm equation is derived and a simple relation is subsequently established between the carbon dose and the remaining trace compound concentration. The results show that the adsorption capacity and relative removal of a trace organic compound at any carbon dose can be estimated directly with the simple equations developed here and data from a single isotherm experiment for the target compound conducted in the natural water of interest. PMID- 17137612 TI - A case study on trace metals in surface sediments and dissolved inorganic nutrients in surface water of Oludeniz Lagoon-Mediterranean, Turkey. AB - Hydro-chemical parameters (salinity, temperature, pH, dissolved oxygen, nitrate, silicate and phosphate) in seawater and major trace metals (Al, Fe, Mn, Cr, V, Zn, Pb, Cu) in sediments were evaluated for the assessment of quality of seawater and sediments in very small lagoon in Mediterranean, Oludeniz. Enrichment factors for metals in sediment were in the range of 1.62-8.09, comparable to crustal rock composition. For metals, comparison with literature data revealed relatively low metal concentrations for Oludeniz sediments. Correlation analyses on the sediment metal data showed strong correlation in between Cr, Fe and Zn. Surface water salinity slightly decreases within the lagoon, indicating that limited fresh waters inflow to the lagoon. In October, the lagoon waters contained very low phosphate concentrations but measurable values of nitrate and silicate, yielding high NO(3)(-)/PO(4)(3-) ratios (90). Very low Chlorophyll-a (biomass indicator) concentrations measured in the lagoon suggest the phosphorus limitation of primary productivity. PMID- 17137613 TI - Hydroxyapatite-supported Ag-TiO2 as Escherichia coli disinfection photocatalyst. AB - A series of hydroxyapatite (HAP), 1wt% Ag-TiO(2) (AT1), 1wt% Ag-HAP and 5wt% AT1/HAP composite catalysts were prepared by incipient wetness and mechanical mixing methods. They were characterized by X-ray diffraction (XRD), FT-IR, SEM and ESCA analyses and their photocatalytic bactericidal activities were measured in suspension using Escherichia coli (E. coli), a water pollutant indicator. The surface analysis revealed that the Ag/Ti ratio is found to be ca. 0.0273 and also it indicated that the titania is present in the form of Ti(4+) and Ag is present as metallic silver. Both the XRD and ESCA analyses confirmed the phase of metallic Ag particles, which played a significant role on the bactericidal activity of the Ag doped TiO(2) catalysts. The FT-IR analysis of HAP revealed that the peak intensity is due to the absorbance of surface PO(4)(3-) group centered at wave number 1030cm(-1) and is drastically decreased upon exposure to UV for 1h. The HAP displayed high amount of bacteria adsorption, ca. 80% during the dark experiments compared to other catalytic systems tested. The cumulative photocatalytic properties of AT1/HAP catalytic system resulted in 100% E. coli bacteria reduction within 2min. PMID- 17137614 TI - Abnormal fragile histidine triad (Fhit) expression in invasive cervical adenocarcinoma: association with tumor aggressiveness. AB - The fragile histidine triad (FHIT) gene is a candidate tumor suppressor gene. Aberrant expression of the encoded protein and inactivation of FHIT correlate with several clinicopathological parameters in various tumor types, including cervical cancer, but Fhit expression has rarely been studied in cervical adenocarcinoma. We assessed Fhit protein expression in 35 surgical specimens of invasive adenocarcinomas of the uterine cervix and investigated whether expression alteration on immunohistochemistry staining is associated with important clinicopathological features. Considerably reduced or absent Fhit staining was observed in 11 cancers (31.4%). By univariate analysis, Fhit protein expression was significantly associated with nodal status (P = .002), histologic grade (P = .000), and International Federation of Gynecology and Obstetrics stage (P = .032). Depth of invasion, tumor size, or parametrial invasion did not show important association with Fhit. Lymph node status, International Federation of Gynecology and Obstetrics stage, and histologic grade are known prognostic factors of cervical adenocarcinoma, and Fhit status on immunohistochemistry staining demonstrated significant association with tumor aggressiveness. Staining of biopsy specimens for Fhit is worthy of study as a prognostic tool. PMID- 17137615 TI - Podocalyxin-like protein 1 expression is useful to differentiate pancreatic ductal adenocarcinomas from adenocarcinomas of the biliary and gastrointestinal tracts. AB - Metastases of adenocarcinomas from the pancreas, liver, and gastrointestinal tract are difficult to distinguish from each other because of their similar morphological and immunohistochemical features. So far, no specific marker for pancreatic ductal adenocarcinomas has been described. Podocalyxin-like protein 1 (PODXL-1) is expressed on vascular endothelium, hematopoietic precursor cells, and renal podocytes. We found that 44% (71/160) of pancreatic ductal adenocarcinomas expressed PODXL-1 in a membranous pattern. There was no expression in intrahepatic cholangiocarcinomas (0/18, P < .001), rarely in adenocarcinomas of the extrahepatic bile ducts (1/13, P = .009), and none in duodenal adenocarcinomas (0/5, P = .070). PODXL-1 expression was seen in only 9% of hepatocellular carcinomas (5/56, P < .001), 9% (4/47, P < .001) of gastric carcinomas, 10% of esophageal adenocarcinomas (2/20, P = .003), and 6% of colonic adenocarcinomas (1/17, P = .001). When used as a differential diagnostic marker, ampullary carcinoma needs to be excluded, as 30% (6/20, P = .24) of ampullary carcinomas stain positive, especially those of the signet-ring type (3/3). Adenocarcinomas of the lung and prostate, and liver metastases of colorectal carcinomas lacked PODXL-1 expression. It is concluded that immunoreactivity for PODXL-1 favors a pancreatic origin if ampullary carcinoma is excluded. PMID- 17137616 TI - The platelet inhibiting effect of a clopidogrel bolus dose in patients on long term acetylsalicylic acid treatment. AB - INTRODUCTION: Addition of clopidogrel to patients treated with ASA has been shown to decrease the incidence of in-stent thrombosis after percutaneous coronary interventions. However, it has also been reported that up to 30% of patients do not achieve adequate platelet inhibition from standard dosages of ASA and clopidogrel. There is a demand for reliable methods to measure the individual platelet inhibiting effect of this combination therapy. MATERIALS AND METHODS: The primary aim of the present investigation was to compare three methods for evaluation of the platelet inhibiting effect of a clopidogrel bolus dose in patients on long-term acetylsalicylic acid treatment. Thirty patients presenting for coronary angiography/PCI were included. Two patients were excluded due to technical problems. All patients were on 75-100 mg ASA/day for at least 8 days. Blood samples were analysed before and 16 h after a 300 mg clopidogrel bolus dose. The platelet inhibiting effect was measured with (1) Whole blood flow cytometry (17 patients); (2) a bed-side test, Platelet Mapping assay for the thrombelastograph (28 patients); and (3) PFA (Platelet function analyser) -100 (26 patients). RESULTS: With flow cytometry, the percentage of platelets expressing P-selectin (p=0.03) on their surface decreased significantly after the bolus dose of clopidogrel. There was also a reduction of platelets binding fibrinogen when stimulated with ADP. A significantly (p=0.002) increased platelet inhibition could also be demonstrated with Platelet Mapping. PFA-100 could not measure any significant platelet inhibiting effect of clopidogrel. CONCLUSION: A significant platelet inhibition could be demonstrated with flow cytometry and the Platelet Mapping assay, but not with PFA-100. However, levels of response for the individual patient with these three methods were inconsistent. Further studies are needed to evaluate how the results correlate to the clinical risk of thrombosis and bleeding. PMID- 17137617 TI - High-risk gestational trophoblastic neoplasia with brain metastases: individualized multidisciplinary therapy in the management of four patients. AB - PURPOSE: To report our recent experience managing four patients with brain metastases of gestational trophoblastic neoplasia (GTN), coordinating systemic chemotherapy with early neurosurgical intervention or stereotactic radiosurgery and intensive supportive care during initial therapy to prevent early mortality. MATERIALS AND METHODS: A series of four consecutive patients with brain metastases from high-risk Stage IV GTN managed at our institution in 2003 and 2005. Patients were assigned FIGO stage and risk score prospectively. Because of concern for chronic toxicity resulting from concurrent moderate dose methotrexate and whole brain radiation, an individualized multidisciplinary approach was used to manage patients. RESULTS: All four women presented with brain and pulmonary metastases; one had multiple liver metastases. Neurological symptoms at presentation included grand mal seizures in 2 patients, left upper extremity hemiparesis and headache each in 1 patient, while 1 patient was asymptomatic. Index pregnancies were term pregnancies in all patients with interval from prior delivery ranging from 2 weeks to 4 years. Two had received prior chemotherapy for postmolar GTN prior to the index pregnancy with incomplete follow-up. Initial hCG values ranged from 26,400 to 137,751 mIU/ml; FIGO risk scores were > or =16 for all patients. Systemic combination chemotherapy was initiated with etoposide and cisplatin followed by moderate/high-dose (500-1000 mg/m(2)) methotrexate combinations. Craniotomy was used before or during the first chemotherapy cycle to extirpate solitary lesions in 3 patients, while stereotactic radiosurgery was administered after the first cycle to treat two brain lesions in the remaining patient. None received whole brain radiation or intrathecal methotrexate. In one patient, selective angiographic embolization was used to control hemorrhage from multiple liver metastases. Two patients required ventilator support early in treatment to allow stabilization from intrathoracic hemorrhage and neutropenic sepsis with respiratory distress syndrome, respectively. Hysterectomy was performed in one patient after completion of salvage chemotherapy. All have completed maintenance chemotherapy and are in prolonged remission (12-24 months). Neurologic sequelae include persistent left upper extremity dyskinesia and weakness in one patient, and episodic grand mal seizures and pseudoseizures in a second patient with a pre-existing seizure disorder. CONCLUSION: This case series documents the utility for a multidisciplinary approach to the treatment of brain metastases from GTN. Using early craniotomy or stereotactic radiosurgery combined with etoposide-cisplatin and moderate/high-dose methotrexate combination chemotherapy, we were able to stabilize patients early in their treatment and avoid whole brain radiation therapy or intrathecal chemotherapy. PMID- 17137619 TI - Improved fertilization and embryo development resulting in birth of live piglets after intracytoplasmic sperm injection and in vitro culture in a cysteine supplemented medium. AB - The effects of cysteine treatment on fertilization rate, intracellular concentration of glutathione, and embryo development in vitro and after embryo transfer were examined following intracytoplasmic sperm injection (ICSI) of in vitro-matured porcine oocytes using a piezo drive unit. Culture of presumed zygotes after ICSI with 1.71-3.71 mM cysteine for 3-12h improved (P<0.05) fertilization rates as compared to treatment with 0.57 mM cysteine or to controls (0mM) (56 to 68%, 48%, 35%, respectively). Extension of treatment time with cysteine beyond 3h did not further increase fertilization rates, suggesting that cysteine promoted early developmental events after ICSI (e.g. decondensation of sperm chromatin). There was no effect of cysteine supplementation on oocyte glutathione levels after ICSI. Pretreatment of spermatozoa for 3h with 1.71 mM cysteine did not improve fertilization rates. The incidence of blastocysts formation when cultured in 1.71 mM cysteine for 3h after ICSI was 31%, which was higher (P<0.05) than controls (18%). Transfer of 20-38 embryos cultured with 1.71 mM cysteine for 3h after ICSI to each of seven recipients yielded three deliveries with an average litter size of 4.0. We concluded that cysteine supplementation for the first 3h after ICSI improved fertilization and embryo development rates, with no influence on glutathione levels in oocytes, and that the cysteine-treated ICSI embryos developed to full term. The study also showed that porcine oocytes matured in a chemically defined medium had the ability for full-term development after piezo-ICSI without additional treatments for oocyte activation. PMID- 17137618 TI - Inhibition of growth of cervical cancer cells using a dominant negative estrogen receptor gene. AB - OBJECTIVE: Estrogen stimulates human papilloma virus oncogene expression, promotes cervical cancer (CC) cell proliferation and prevents apoptosis. Therefore, blockage of estrogen function may have therapeutic application to CC. METHODS: CasKi CC cells were transfected with an adenovirus expressing a dominant negative estrogen receptor gene (Ad-ER-DN) and their responses were investigated by RT-PCR, Flow Cytometry and Western blot assays. RESULT: Transfected cells showed disturbance of cell colony morphology, reduced HPV E6 and E7 mRNA, interruption of cell proliferation, reduced cyclin D1 protein and expression of apoptosis. CONCLUSION: We report, for the first time, the use of Ad-ER-DN to block estrogen receptors which led to dramatic changes in CC cells that are consistent with the possible reactivation of cellular p53 and Rb function. Their reactivation most likely allowed the recognition of existing chromosome abnormalities as a serious stress signal and the initiation of a cascade of cellular events in response to the stress, including the activation of the core apoptotic machinery which led to self-destruction of the CC cells. PMID- 17137620 TI - Development of a novel CASA system based on open source software for characterization of zebrafish sperm motility parameters. AB - Although computer-assisted sperm analysis (CASA) outperforms manual techniques, many investigators rely on non-automated analysis due to the high cost of commercial options. In this study, we have written and validated a free CASA software primarily for analysis of fish sperm. This software is a plugin for the free National Institutes of Health software ImageJ and is available with documentation at . That it is open source makes possible external validation, should improve quality control and enhance the comparative value of data obtained among laboratories. In addition, we have improved upon the traditional velocity straight line (VSL) algorithm, eliminating inaccurate characterization of highly curved fish sperm paths. Using this system, the motion of zebrafish (Danio rerio) sperm was characterized relative to time post-activation and the impact of acquisition conditions upon data analysis determined. There were decreases in velocity and path straightness (STR), but not linearity (LIN), relative to time. From 30 to 300 frames/s, frame rate significantly affected curvilinear velocity (VCL) and STR measurements. Sperm density in the field of view did not affect any measured parameter. There was significant inter-male variation for VCL, VSL, velocity average path (VAP), percent motility, path character (STR, LIN), and duration of motility. Furthermore, relative sperm output (a measure reflecting both semen volume and concentration) was positively correlated to percent motility. For all motion parameters measured (except duration), the average CV was < or =10%, comparable to values obtained using commercial systems. PMID- 17137621 TI - A mathematical model for simulating the bone remodeling process under mechanical stimulus. AB - OBJECTIVES: Among the current mathematical models for bone remodeling, few can consider bone resorption due to overload. The objective of this paper is to develop a new bone remodeling model which can simulate both underload and overload resorptions that often occur in dental implant treatments. METHODS: Based on the traditional model, a new mathematical equation relating the density change rate with mechanical stimulus has been developed. The new equation contains an additional quadratic term which can produce reduction in bone density at high load levels. In addition, to fully exploit the characteristics of this model, a range of different bone remodeling behaviors were studied under the load cases with both constant and varying stress magnitudes. Finally, the model was applied in conjunction with the finite element method to a practical case of dental implant treatment. RESULTS: The FE analysis results showed that bone resorption at the neck of the implant occurred due to occlusal overload but then resorption stopped after some time before reaching the coarse threads. Meanwhile, the density of the bone deeper into the mandible increased slightly due to the additional mechanical stimulus provided by the occlusal load. This phenomenon is observable in some clinical situations. SIGNIFICANCE: The new model can describe the bone overload resorption, a feature which is absent in most of the current models. And by simulating the dental implant treatment using FE method, the ability of the new mathematical model to simulate overload bone resorption has been clearly demonstrated. PMID- 17137622 TI - Matrix metalloproteinase-9 gene -1562C/T polymorphism mitigates preeclampsia. AB - Although the aetiology of preeclampsia is unknown, there is substantial evidence that it finds its roots in abnormal placentation. Prerequisites for successful placentation include trophoblast invasion, degradation and remodelling of the uterine decidual extracellular matrix, and apoptosis without thrombosis. We tested this hypothesis by analysing the effect of functional polymorphisms in the genes coding for MMP9, MMP3 and annexin A5 on the risk of preeclampsia using a case-control design. In 163 women with preeclampsia and 163 controls we studied the association with polymorphisms in the MMP9 (-1562 C/T), MMP3 (-1612 5A/6A) and annexin A5 (-1 C/T) genes using logistic regression analysis. A lower prevalence of the rare T allele of the MMP9 (-1562 C/T) polymorphism in women with preeclampsia was found (odds ratio 0.48, 95% confidence interval 0.25-0.90). The distribution of the MMP3 (-1612 5A/6A) and annexin A5 (-1 C/T) gene polymorphisms were similar in cases and controls. Our results suggest that the MMP9-1562T allele is associated with a reduced risk of preeclampsia and therefore may protect against maladaptation of the spiral arteries and decreased decidual degradation. The elevated MMP9 concentrations reported to be associated with the 1562T allele might be essential for the development of an adequate maternal-fetal interface early in pregnancy by facilitating trophoblast apoptosis and degradation. PMID- 17137623 TI - Targeting of calcium:calmodulin signals to the cytoskeleton by IQGAP1. AB - Mast cells reorganize their actin cytoskeleton in response to cytosolic calcium signals while in parallel secreting histamine and other inflammatory mediators. The effect of calcium on actin is mediated in large part through calmodulin. EGFP tagged calmodulin is concentrated in the actin-rich cortex of RBL-2H3 mast cells. Transfection with small interfering RNA directed against the actin and calmodulin binding protein IQGAP1 dramatically reduced expression of the latter protein and reduced or eliminated the concentration of calmodulin at the actin-rich cortex. Both actin reorganization and secretion were enhanced in IQGAP1 knockdown cells. Our results suggest a model in which calmodulin is targeted to and sequestered at the actin cytoskeleton by IQGAP1. Upon cell stimulation and the subsequent [Ca2+]i increase, it is immediately available to activate local downstream targets. PMID- 17137624 TI - Simultaneous diagnosis of CD3+ T-cell large granular lymphocyte leukaemia and true thymic hyperplasia. AB - CD3+ T-cell large granular lymphocyte (LGL) leukaemia is a mature T-cell neoplasm of cytotoxic T-lymphocytes. Neutropenia represents the most frequent peripheral blood cytopenia associated with CD3+ T-cell LGL leukaemia. A wide variety of diseases associated with LGL leukaemia have been reported, both autoimmune and neoplastic. We describe for the first time the association of true thymic hyperplasia with CD3+ T-cell line LGL leukaemia. The patient presented with severe symptomatic neutropenia. Complete and persistent haematological and molecular remission was induced with an association of low-dose methotrexate and cyclosporin A, followed by thymectomy. PMID- 17137625 TI - Advances in AML immunotherapy: the case for T-bodies. PMID- 17137626 TI - Functional analysis of ori1 and repA of the R-plasmid pSJ5.6 from Neisseria gonorrhoeae. AB - The functional ori1 of the 5.6kb gonococcal R-plasmid pSJ5.6 contains an A-T rich region followed by four 22bp direct repeats and one 19bp inverted repeat. The replication region of the plasmid also contains a gene encoding for a 39kD RepA protein. We have further assessed the functionality of the replication region in pSJ5.6, an-iteron type plasmid, using in vivo complementation assays in Escherichia coli. A 2.1kb PstI-RsaI fragment containing the ori1 and repA gene of pSJ5.6 was cloned into vector pZErO -2 to obtain pZA-MRR. The pUC origin in pZA MRR was deleted to render the plasmid dependable on the cis-acting ori1 for replication. The resulting plasmid, pMRR, was capable of replication and maintenance in E. coli. We also cloned the ori1 and repA gene separately to obtain pA-Ori and pZG-Rep, respectively. Using in vivo complementation assays, we demonstrated that the ori1(+) plasmid (pA-Ori) was maintained only when the RepA protein was supplied in trans by the high copy number plasmid pZG-Rep. PMID- 17137627 TI - Toxicity of microcystins in the isolated hepatocytes of common carp (Cyprinus carpio L.). AB - The toxicity of hepatotoxic microcystins produced mainly by Microcystis aeruginosa in mammals and fishes was well studied in recent years. However, there were scarcely reports in toxic effects of microcystins on isolated hepatocytes of fishes, especially investigation of microcystin-induced apoptosis and/or necrosis in carp hepatocytes. In the present study, the isolated hepatocytes of common carp were exposed to various concentrations of microcystins (0.01, 0.1, 1, 10, 100, 1000 microg L(-1)) for 2, 4, 8, 16 and 24h, respectively, and cytotoxicity of microcystins in the toxin-treated cells was determined. Results of this study showed that cytotoxicity of microcystins on carp hepatocytes was time and dose dependent, and the approximate LC(50) of microcystins in carp hepatocytes was 169.2 microg L(-1). The morphological changes typical of apoptosis, such as blebbing of cell membrane, condensation and fragmentation of cell nucleus were observed in the hepatocytes exposed to microcystins (1, 10 and 100 microg L(-1)) using fluorescence and differential interference contrast microscopy. Agarose gel electrophoresis of DNA demonstrated a typical apoptotic "ladder pattern" in microcystin-treated hepatocytes after 16 h of exposure. Results of the present study indicated that the form of cell death in microcystin-treated hepatocytes depend on the exposure dose of toxin. When lower concentration of microcystins (10 and 100 microg L(-1)) was used for exposure, carp hepatocytes died in apoptosis while, when higher one used (1000 microg L(-1)), they died in the form of necrosis. PMID- 17137628 TI - Genetic variability in Daphnia magna and ecotoxicological evaluation. AB - The fresh water crustacean Daphnia magna is widely used as a test organism in aquatic toxicology to assess the adverse effects of individual substances or complex mixtures, e.g. industrial wastewaters. Cultures are held in several European testing laboratories and testing is typically carried out according to internationally standardised protocols. However, despite accounting for many potential confounding factors these guidelines do not currently take into account any specification related to the use of a specific clone. Cultures from seven laboratories were used to assess genetic variability by random-amplified polymorphic DNA polymerase chain reaction. Results pointed out the existence of two main clone clusters Responses in the acute Daphnia immobilisation test showed no direct correlation with genetic clusters resulting from random genetic markers (random-amplified polymorphic DNA) analysis. Considering that genetic differences are the most probable cause for the ecotoxicological test data, further analysis concerning gene expression and genetic stability should be performed. PMID- 17137629 TI - Comparison and analysis of different approaches for estimating the human exposure to phthalate esters. AB - The human exposure estimates for dibutyl (DBP) and bis(2-ethylhexyl) phthalate (DEHP) made by two models EUSES and ACC-Human, and by an estimation approach which utilized measured concentrations in exposure media, were compared. The approach which utilized the latest monitoring data for important exposure media, yielded median daily intakes for adult humans for DBP and DEHP of 2.7 and 5.6 microg/kg body weight per day, respectively, which were in the same range as previous estimates based on back-calculation from urinary metabolites. EUSES estimated average daily intakes of DBP and DEHP for humans that were between 8 and 13 times lower. ACC-Human does not estimate average daily intakes, but ACC Human-estimated human milk concentrations/fugacities were more than a thousand times lower than measured concentrations/fugacities in human milk. It was concluded that the two models underestimate human exposure to phthalate esters because they consider only a few key pathways that are known to be important for other, more persistent, hydrophobic organic compounds. Further, it was shown that there are differences between the two models on the methodology for estimating concentrations in exposure media such as vegetation, milk, beef and fish. ACC Human uses a mechanistic approach for estimating transfer through aquatic and terrestrial food chains that are known to be important for human exposure to persistent, hydrophobic organics and can, unlike EUSES, account for food chain metabolism. It proved difficult, however, to obtain organism metabolism rates needed as model inputs to ACC-Human. If exposure estimates of phthalate esters are needed, it is recommended to use an estimation approach based on high quality monitoring data as presented here and/or back-calculate daily intake from concentrations of metabolites in human urine samples from the general population. PMID- 17137630 TI - Roles of distal arginine in activity and stability of Coprinus cinereus peroxidase elucidated by kinetic and NMR analysis of the Arg51Gln, -Asn, -Leu, and -Lys mutants. AB - In heme peroxidases, a distal His residue plays an essential role in the initial two electron oxidation of resting state enzyme to compound I by hydrogen peroxide. A distal Arg residue assists in this process. The contributions of the charge, H-bonding capacity, size, and mobility of this Arg residue to Coprinus cinereus peroxidase (CIP) reactivity and stability have been examined by substituting Arg51 with Gln (retains H-bond donor at N epsilon position), Asn (small size, H-bond donor and acceptor), Leu (similar to Asn, but hydrophobic), and Lys (charge and H-bond donor, but at N zeta position). UV-visible spectroscopy was used to monitor pH-linked heme changes, compound I formation and reduction, fluoride binding, and thermostability. (1)H NMR spectroscopy enabled heme pocket differences in both resting and cyanide-ligated states of the enzymes to be evaluated and compared with wild-type CIP. We found that the H-bonding capacity of distal Arg is key to fast compound I formation and ligand binding to heme, whereas charge is important for lowering the pK(a) of distal His and for the binding and stabilisation of anionic ligands at heme iron. The properties of the distal Arg residue in CIP, cytochrome c peroxidase (CCP) and horseradish peroxidase (HRP) differ significantly in their pH induced transitions and dynamics. PMID- 17137631 TI - Considerations on influence of charge distribution on determination of biomolecules and microorganisms and tailoring the monolithic (continuous bed) materials for bioseparations. AB - The importance of continuous beds (monoliths) as separation materials is connected with their better chromatographic properties and easier preparation in comparison to particulate-packed columns. Moreover the tuning of porosity as well as surface chemistry can lead to obtaining of highly selective materials, especially useful in separation of biologically important compounds or even microorganisms. To obtain high selectivity for such analytes as e.g. proteins, it is often important to have a knowledge about their shape, size, charge and finally charge distribution. This article presents our considerations on the charge distribution on the monolithic stationary phase and surface of such species as proteins or microorganisms as well as its eventual influence on the separation or sample preparation processes and tuning of their selectivity. PMID- 17137632 TI - The essence of DNA sample preparation for MALDI mass spectrometry. AB - Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry has become an important analytical technique in nucleic acid research. MALDI is used for quality control of oligonucleotides as well as for analyzing DNA markers. Sample preparation of nucleic acids is crucial for obtaining high-quality mass spectra. Sample purity, solvent content, suitable matrices, and substrate surfaces, as well as laboratory conditions affect spectra quality. This review presents essential information with regard to sample preparation, DNA modification chemistry, and DNA purification, along with a discussion of instrumental advances, which facilitate and extend the applicability of MALDI in genomics. PMID- 17137633 TI - Detecting correlation changes in electrophysiological data. AB - A correlation multi-variate analysis of variance (MANOVA) test to statistically analyze changing patterns of multi-electrode array (MEA) electrophysiology data is developed. The approach enables us not only to detect significant mean changes, but also significant correlation changes in response to external stimuli. Furthermore, a method to single out hot-spot variables in the MEA data both for the mean and correlation is provided. Our methods have been validated using both simulated spike data and recordings from sheep inferotemporal cortex. PMID- 17137634 TI - Predicting domain-specific insight of schizophrenia patients from symptomatology, multiple neurocognitive functions, and personality related traits. AB - This study examines the contribution of various neurocognitive functions, clinical characteristics, and personality traits to the prediction of three insight dimensions. Clinically stable schizophrenia patients (n=107) residing in the community were evaluated using the Positive and Negative Syndrome Scale, the Scale for the Assessment of Unawareness of Mental Disorder, and a comprehensive battery of instruments to measure personality related variables and neurocognitive functioning. Step-wise multivariate regression analysis indicates significant association of variability in insight dimensions with neurocognitive functioning (20-41%), personality related traits (8-18% temperament factors, 4-7% self-constructs, 10-14% coping styles), severity of symptoms (about 7%), illness duration (6%), and education (about 5%). Poor insight was attributed to impairment in visual and movement skills, sustained attention, executive functions, intensity of autistic preoccupations and positive symptoms, as well as increased novelty seeking behavior, task and emotion oriented coping styles, better self-esteem, self-efficacy, and higher education. Better awareness was related to better performance of neurocognitive tasks, reward dependence behavior, avoidant coping style, and longer illness duration. Aside from common indicators for the various insight dimensions, we defined specific indicators for each insight dimension. Thus, insight dimensions in schizophrenia patients residing in the community were attributed to neurocognitive and personality related factors rather than to psychopathological symptoms. The findings enable better understanding of the multifactorial nature of insight and highlight targets for more effective intervention and rehabilitation. PMID- 17137635 TI - Identification of MCP/CD46 analogue on bovine erythrocytes using the new monoclonal antibody IVA-520. AB - MCP/CD46 is a widely distributed C3b/C4b binding regulatory glycoprotein of the complement system that has been identified on all human peripheral blood cells except erythrocytes. In this paper, we describe the identification of bovine CD46 on all blood cells, including erythrocytes, with the newly prepared monoclonal antibody IVA-520. This antibody cross-reacts with human and pig cells. Furthermore, the molecule identified by IVA-520 functionally behaves as the MCP molecule, showing cofactor activity for the factor I-mediated cleavage of bovine C3 complement factor. PMID- 17137636 TI - Genetic analysis of intracranial tumors in a murine model of glioma demonstrate a shift in gene expression in response to host immunity. AB - For the study of malignant glioma, we have previously characterized a highly tumorigenic murine astrocytoma, SMA-560, which arose spontaneously in an inbred, immunocompetent VM/Dk mouse. Using this cell line as a model of murine glioma, we performed DNA microarray analysis of autologous normal murine astroctyes (NMA) and SMA-560 tumor cells grown in monolayer culture or intracranially in syngeneic immunocompetent or immunocompromised hosts in order to determine whether tumors grown in vitro recreate the complex genetic regulation that occurs in vivo. Our findings support our hypothesis that glioma phenotype in vitro may be quite different in vivo and significantly altered by in situ growth factors and other invading cell populations. PMID- 17137637 TI - Pros and cons of treating murine myasthenia gravis with anti-C1q antibody. AB - To test the feasibility of classical complement pathway manipulation in experimental autoimmune myasthenia gravis (EAMG) treatment, C57BL/6 (B6) and RIIIS/J mice with EAMG were treated with 10 microg or 100 microg of anti-C1q Ab or isotype Ab. Treatment with 10 microg anti-C1q Ab significantly reduced the clinical severity, decreased lymph node cell IL-6 production and T cell populations. Conversely, administration of 100 microg anti-C1q Ab caused harmful side effects such as increased serum anti-acetylcholine receptor antibody, immune complex, C3 and lymph node B cell levels and kidney C3 and IgG deposits, which reduced the treatment efficacy. PMID- 17137638 TI - Effects of macrolides on antigen presentation and cytokine production by dendritic cells and T lymphocytes. AB - Macrolides are effective therapeutic agents for chronic respiratory tract diseases, such as chronic sinusitis, sinobronchial syndrome and diffuse panbronchiolitis. Although only limited information is available about their mechanisms, suppression of various inflammatory cytokines (IL-8, etc.) and some transcription factors has been reported to be involved. Non-typeable Haemophilus influenzae (NTHI) is one of the most important pathogens of the respiratory tract. P6 is one of the outer membrane proteins of NTHI and the target antigen of protective antibodies. To analyze the influence of macrolides on human dendritic cells (DCs), we treated DCs with macrolides and used them as antigen-presenting cells (APCs). Clarithromycin, roxithromycin and prednisolone suppressed the in vitro proliferative response of CD4+ T cells to P6 and also the production of cytokines. As a control, we also cultured DCs alone and exposed them to the medicament, while conversely culturing T cells without adding any drugs to the cultures. The results showed similar tendencies for suppression of immune responses. These findings suggest that macrolides suppress the antigen-specific immune responses of DCs in vitro. PMID- 17137639 TI - Voice therapy in pediatric functional dysphonia: a prospective study. AB - OBJECTIVE: We evaluated the efficacy of voice therapy according to Borragan's method associated to S. Magnani's vocal counselling in functional dysphonia in children. PATIENTS AND METHODS: We prospectively treated 16 patients with vocal fold nodules (10 males, 6 females). Age ranged from 6 to 11 years with a mean age of 9 years. We performed a full screening phoniatric evaluation. In addition psychological tests were carried out to investigate psychological background. RESULTS: We lost three patients at follow-up; one patient received surgery, eight patients healed (43.75%), four improved (25%). There was no statistical difference in the analysis of electroacoustical parameters while MPT significatively raised after therapy. CONCLUSION: If patients have motivation voice therapy could improve functional dysphonia in children. It is also important psychological background. Further studies on bigger populations with long-term follow-up are needed. PMID- 17137640 TI - Choking on pins, needles and a blowdart: aspiration of sharp, metallic foreign bodies secondary to careless behavior in seven adolescents. AB - OBJECTIVE: To review the demographics, pathophysiology and management of aspiration of sharp, metallic foreign bodies; to review prevention strategies. DESIGN: Case series and review of the related literature (1932-2006). SETTING: Tertiary care pediatric hospital. PATIENTS: All patients presenting to BC Children's Hospital with aspiration of sharp, metallic foreign bodies since 1998. RESULTS: The seven patients ranged in age from 11 to 15 years (mean 13 years). The two boys had been playing with other boys at the time of aspiration. One aspirated a thumbtack and one aspirated a homemade blowdart (which traveled from his carina to his subglottis with coughing). Three of the five girls had been holding thumbtacks or a darting pin between their lips while putting up posters or sewing. The other two girls, who aspirated thumbtacks, refused to provide a history. One was treated successfully 6 days after the aspiration. The other girl withheld the history for over a year, presented with hemoptysis and eventually required thoractomy and right main bronchotomy to remove the tack. All of the other patients were diagnosed and successfully treated by rigid bronchoscopy within 24h. (Two patients initially had a failed attempt at foreign body removal by flexible bronchoscopy at another hospital.) This is the second largest series of aspiration of sharp, metallic foreign bodies since at least 1966. Adolescents are still aspirating sharp, metallic foreign bodies because of the same behaviors described by Jackson and Jackson in 1932. Education about choking hazards is needed for this age group. CONCLUSIONS: Aspiration of a sharp, metallic foreign body is a serious injury which is best treated by rigid bronchoscopy. These injuries potentially could be prevented through education. PMID- 17137641 TI - Development of a non-denaturing electrophoresis system for characterization of neutralizing epitopes on HPV virus-like particles. AB - The precise structure of the HPV16 major neutralizing epitope recognized by H16.V5 monoclonal antibody is unknown. This paper describes a novel polyacrylamide gel electrophoresis (PAGE) for separation of HPV virus-like particles (VLPs) using cetyltrimethylammonium chloride (CTAC) as a solubilizing agent. CTAC PAGE employs KOH/CH3CO2H (pH 4-5.4) as a buffer system, K+ as the leading ion and 3-aminopropionic acid as a trailing ion. The unique characteristics of a cationic electrophoresis system allow separation of VLPs without heat denaturation. HPV VLP gel migration patterns were dependent on pre treatment conditions: (1) thiol-agent reduction alone resulted in a 174 kDa band (interpreted as a L1 trimer), a 53 kDa band (size of the L1 monomer), as well as higher Mr aggregates consistent with a pentamer size; (2) both heat denaturation and thiol-agent reduction resulted in a 53 kDa band. Western blot analysis showed that the 174 kDa L1 trimer was strongly immunoreactive with H16.V5 and HPV16 VLP ELISA positive human sera, whereas no reactivity was seen with the monomeric L1 unit. These data suggest that a structure consistent with the migration pattern of a L1 trimer contains the major neutralizing epitope recognized by the H16.V5 MAb and human sera. PMID- 17137642 TI - New insights into corollary discharges mediated by identified neural pathways. AB - Sensory systems respond not only to stimuli from the environment but also to cues generated by an animal's own behaviour. This leads to problems of sensory processing because self-generated information can occur at the same time as external sensory information. However, in motor regions of the CNS corollary discharges are generated during behaviour. These signals are not used to generate movements directly but, instead, interact with the processing of self-generated sensory signals. Corollary discharges transiently modulate self-generated sensory responses and can prevent self-induced desensitization or help distinguish between self-generated and externally generated sensory information. Here, we review recent work that has identified corollary discharge pathways at different levels of the CNS in vertebrates and invertebrates. PMID- 17137643 TI - Redox regulation of neuronal survival mediated by electrophilic compounds. AB - The importance of phosphorylation of key threonine, serine and tyrosine residues is a well known essential feature of many signal transduction pathways. A similar, highly conserved redox reaction involving cysteine thiols is now emerging as an important regulator of protein function. An example of this redox regulation is S-nitrosylation (the transfer of a nitric oxide group to a key protein thiol). Here, we review the chemical biology of an additional class of drugs, electrophiles (electron-deficient carbon centers), that react with key protein thiols, and provide insights into a broader class of reactions implicated in redox signaling. Interestingly, certain electrophilic compounds, including endogenous metabolites and natural products, seem to have neuroprotective effects, and this has resulted in the development of neuroprotective electrophilic drugs, including prostaglandin derivatives and hydroquinones, that exert their action through activating antioxidant-signaling cascades. PMID- 17137644 TI - Characterization of drug-resistant recombinant influenza A/H1N1 viruses selected in vitro with peramivir and zanamivir. AB - There is a limited information with regard to the neuraminidase (NA) mutations conferring resistance to peramivir and zanamivir in the influenza N1 background. In this study, an influenza A/WSN/33 (H1N1) recombinant virus was passaged under peramivir or zanamivir pressure. The peramivir-selected variant had a H274Y mutation in the neuraminidase (NA) gene conferring resistance to peramivir and oseltamivir but susceptibility to zanamivir. The zanamivir-selected variant had a massive deletion in the region encoding the NA active center and an A200T hemagglutinin mutation. This variant exhibited reduced susceptibility to zanamivir with a drug-dependent phenotype. PMID- 17137645 TI - Discrimination learning and reversal of the conditioned eyeblink reflex in a rodent model of autism. AB - Offspring of rats exposed to valproic acid (VPA) on gestational day (GD) 12 have been advocated as a rodent model of autism because they show neuron loss in brainstem nuclei and the cerebellum resembling that seen in human autistic cases . Studies of autistic children have reported alterations in acquisition of classical eyeblink conditioning and in reversal of instrumental discrimination learning . Acquisition of discriminative eyeblink conditioning depends on known brainstem-cerebellar circuitry whereas reversal depends on interactions of this circuitry with the hippocampus and prefrontal cortex. In order to explore behavioral parallels of the VPA rodent model with human autism, the present study exposed pregnant Long-Evans rats to 600 mg/kg VPA on GD12 and tested their offspring from Postnatal Day (PND26-31) on discriminative eyeblink conditioning and reversal. VPA rats showed faster eyeblink conditioning, consistent with studies in autistic children . This suggests that previously reported parallels between human autism and the VPA rodent model with respect to injury to brainstem cerebellar circuitry are accompanied by behavioral parallels when a conditioning task engaging this circuitry is used. VPA rats also showed impaired reversal learning, but this likely reflected "carry-over" of enhanced conditioning during acquisition rather than a reversal learning deficit like that seen in human autism. Further studies of eyeblink conditioning in human autism and in various animal models may help to identify the etiology of this developmental disorder. PMID- 17137646 TI - Distinct activities of the related protein kinases Cdk1 and Ime2. AB - In budding yeast, commitment to DNA replication during the normal cell cycle requires degradation of the cyclin-dependent kinase (CDK) inhibitor Sic1. The G1 cyclin-CDK complexes Cln1-Cdk1 and Cln2-Cdk1 initiate the process of Sic1 removal by directly catalyzing Sic1 phosphorylation at multiple sites. Commitment to DNA replication during meiosis also appears to require Sic1 degradation, but the G1 cyclin-CDK complexes are not involved. It has been proposed that the meiosis specific protein kinase Ime2 functionally replaces the G1 cyclin-CDK complexes to promote Sic1 destruction. To investigate this possibility, we compared Cln2-Cdk1 and Ime2 protein kinase activities in vitro. Both enzyme preparations were capable of catalyzing phosphorylation of a GST-Sic1 fusion protein, but the phosphoisomers generated by the two activities had significantly different electrophoretic mobilities. Furthermore, mutation of consensus CDK phosphorylation sites in Sic1 affected Cln2-Cdk1- but not Ime2-dependent phosphorylation. Phosphoamino acid analysis and phosphopeptide mapping provided additional evidence that Cln2-Cdk1 and Ime2 targeted different residues within Sic1. Examination of other substrates both in vitro and in vivo also revealed differing specificities. These results indicate that Ime2 does not simply replace G1 cyclin-CDK complexes in promoting Sic1 degradation during meiosis. PMID- 17137647 TI - Persistent left superior vena cava: multi-slice CT images and report of a case. AB - Abnormalities of the vena cava system are usually asymptomatic and discovered incidentally during catheter placement or pacemaker implantation. Persistent left superior vena cava (PLSVC) is caused by failure of involution of the left anterior cardinal vein caudal to the left brachiocephalic vein during embryonic development. It is a benign condition, but becomes dangerous during pacemaker lead implantation, especially in emergency situations and when the right superior vena cava is absent. This is brought about by difficulty in pacemaker lead maneuvering into the right ventricle. A 64-cut multi-slice computed tomographic (MSCT) scan can show clear spatial relationship of the heart with its surrounding structures. We present a case of PLSVC discovered during pacemaker implantation, and viewed by 64-cut MSCT scan. PMID- 17137648 TI - Sequential measurements of troponin Ic values in brain-dead patients considered as potential heart donors. AB - It was suggested that a single value of normal or increased plasma cardiac troponin T or I (cTnT or cTnI) concentration could contribute to estimate donor myocardial damage and function in brain-dead patients. In patients with acute coronary syndromes, an initial normal value of troponin must be confirmed several hours later but no such recommendations exist for brain-dead patients. We investigated the relationship between two sequential (6 h interval) measurements of plasma cTnI concentrations in brain-dead patients considered as potential heart donors. The first and the second TnIc values were correlated with an adjusted r2 value of 0.92 (p<0.001). Our results suggest therefore that it is not necessary to repeat the measurements, when the value of plasma cTnI concentration is taken into consideration in the algorithm for cardiac harvesting. PMID- 17137649 TI - Preoperative determinants of recovery time in adult Fallot patients after late pulmonary valve replacement. PMID- 17137650 TI - Short-term progress of patients with self-limited first episode atrial fibrillation. AB - Clear guidelines are lacking for the management of the clinical pattern of first episode type atrial fibrillation (AF). We retrospectively analysed the clinical evolution of patients with self-limited first episode AF identified from among from 200 patients who had been hospitalised in our cardiology ward with AF. Of the 200 patients, 33 (16.5%) were self-limited first episode. Over a mean follow up of 19.5+/-12.5 months (53.6 patient-years), 7 patients (21%) had recurrence of arrhythmia and 4 patients (12%) had a thromboembolic episode (7.4 episodes/100 patient-years of follow-up). These results indicate that the profile of thromboembolic risk, and not that of clinical profile presentation of AF, should be the criteria by which to judge the indication for anti-coagulant treatment. PMID- 17137652 TI - Increased beat-to-beat QRS amplitude variability in patients with congestive cardiac failure. AB - In patients with ischemic heart disease, dobutamine provoked-stress increases beat-to-beat QRS amplitude variability, which may be due to an electrical instability of the myocardium. Our results in this pilot study showed that patients with congestive heart failure had significantly higher beat-to-beat QRS amplitude variability compared to controls in resting supine posture. This may partly be due to myocardial disease or irregular respiration in this patient group. PMID- 17137654 TI - Long-term survival with partial atrioventricular septal defect. PMID- 17137655 TI - Cardiovascular risk stratification by means of the SCORE system in health care workers in Veracruz, Mexico. AB - BACKGROUND: To assess cardiovascular disease risk by means of the SCORE system (Systematic Coronary Risk Evaluation) in health care workers (HCW) from Veracruz, Mexico. METHODS: A cross-sectional study was undertaken at the Mexican Institute of Social Security from Veracruz. Seventy six HCW without physical limitations (NYHA Functional Class I) were included. All HCW answered a standardised medical history questionnaire and were evaluated by physical examination and lab tests. The cardiovascular risk was assessed through the SCORE system. RESULTS: The median age of participants was 47 years (90% range 42-57 years), female HCW had higher prevalence of obesity and lower prevalence of overweight compared to male HCW (52% vs 23% for obesity and 26% vs 63% for overweight; p=0.014). The prevalence of hypertension was 22%, type 2 diabetes 8%, hypercholesterolemia 70%, hypertriglyceridemia 47% and mixed hyperlipidemia 26%. Cardiovascular risk assessed by the SCORE system showed that 14% of all patients had a cardiovascular risk higher than 2% and 51% had a risk lower than 1%. CONCLUSIONS: In this first study of cardiovascular risk assessment by means of the SCORE system in HCW in Mexico, we found that 14% of them have a cardiac risk higher than 2% and that it is double than expected for their age but it is lower than reported in a European population. Also, we found a higher prevalence of hypercholesterolemia and mixed hyperlipidemia showing poor education and treatment for cardiovascular prevention. PMID- 17137656 TI - Effects of smoking on myocardial infarction at early ages. PMID- 17137657 TI - Long-term and short-term alcohol consumption and levels of C-reactive protein. AB - The association between alcohol consumption and CRP was evaluated in a longitudinal study among 72 middle aged adults for whom 572 repeated measurements of CRP were obtained. Short- and long-term alcohol consumption were analyzed using moving windows of average alcohol consumption levels to establish a dose response relationship. To control for intra-individual dependence of measurements, generalised estimating equations were used. Average alcohol consumption during the last 12 months showed a clear U-shaped relationship with CRP-levels, with lowest levels found for moderate alcohol consumption of less than 16 g/day. For short-term alcohol consumption no association was found. PMID- 17137658 TI - Ventricular fibrillation due to severe mitral valve prolapse. AB - Mitral valve prolapse usually has a good prognosis. However, an association between mitral valve prolapse and atrial and ventricular arrhythmias has been described. This case presents a patient who was admitted after cardiac resuscitation due to ventricular fibrillation. A severe mitral valve prolapse was the only pathology found. PMID- 17137659 TI - Successful catheter ablation of idiopathic left ventricular tachycardia during sinus rhythm. AB - Successful catheter ablation of idiopathic left ventricular tachycardia (ILVT) can be performed at the site of the earliest Purkinje potential or at the site with recording of diastolic and presystolic Purkinje potentials simultaneously during ventricular tachycardia (VT). However, these critical potentials might be difficult to be recorded and mapped in some patients during VT. It is rare to report the ablation of ILVT during sinus rhythm. We present a case with ILVT who received successful catheter ablation during sinus rhythm. PMID- 17137660 TI - Interpretation of California mastitis test scores using Staphylococcus aureus culture results for screening of subclinical mastitis in low yielding smallholder dairy cows in the Dar es Salaam region of Tanzania. AB - Screening of subclinical mastitis under field conditions is done using the California mastitis test (CMT). CMT score of > or = 1 corresponding to > or = 500,000 somatic cells ml(-1) is commonly used as threshold of subclinical mastitis in temperate countries. However, given the innately high physiological level of somatic cells in low yielding dairy cows, this threshold may not apply to low yielding dairy cows. The current study was undertaken to investigate the clinical utility of CMT for screening of Staphylococcus aureus subclinical mastitis in low yielding smallholder dairy cows in Tanzania. A total of 1151 of quarter-milk samples were CMT tested, of these 914-originated from cows with a lactation period of 14-305 days. All samples were screened for subclinical mastitis by the CMT as well as microbiological culture of single, duplicate (two consecutive) and triplicate (three consecutive) samples as a gold standard. For the duplicate and triplicate quarter-samples, cows were considered positive for S. aureus subclinical mastitis if results of microbiologic culture for S. aureus were positive for two of two, and for at least two of the first three consecutive quarter-milk samples collected from that cow, respectively. Using a CMT score of > or = 1 would classify 78.6% of the 940 quarter-samples as positive. Eighty-two percent of the samples in which S. aureus was isolated had CMT scores > or = 2; this would classify 51.6% of the 940 quarter-samples as positive. For the single sample, this cut-off had sensitivity, specificity and likelihood ratio for S. aureus of 0.87, 0.83 and 4.24, respectively. For the duplicate quarter-milk samples this cut-off had sensitivity, specificity, and likelihood ratio of 0.94, 0.86, and 5.19. While, for the triplicate quarter-milk samples this cut-off had sensitivity, specificity and likelihood ratio of 0.97, 0.92 and 7.47, respectively. Based on these results and practical considerations, it is concluded that CMT score of > or = 2 corresponding to > or = 800,000 somatic cells Ml(-1) is the best cut-off to correctly identify S. aureus intramammary infections in low yielding dairy cows in Tanzania. PMID- 17137661 TI - Enrichment of functional redox reactive proteins and identification by mass spectrometry results in several terminal Fe(III)-reducing candidate proteins in Shewanella oneidensis MR-1. AB - Identification of the proteins directly involved in microbial metal-reduction is important to understanding the biochemistry involved in heavy metal reduction/immobilization and the ultimate cleanup of DOE contaminated sites. Although previous strategies for the identification of these proteins have traditionally required laborious protein purification/characterization of metal reducing capability, activity is often lost before the final purification step, thus creating a significant knowledge gap. In the current study, subcellular fractions of Shewanella oneidensis MR-1 were enriched for Fe(III)-NTA reducing proteins in a single step using several orthogonal column matrices. The protein content of eluted fractions that demonstrated activity was determined by ultra high pressure liquid chromatography coupled with tandem mass spectrometry (LC MS/MS). A comparison of the proteins identified from active fractions in all separations produced 30 proteins that may act as the terminal electron-accepting protein for Fe(III)-reduction. These include MtrA, MtrB, MtrC and OmcA as well as a number of other proteins not previously associated with Fe(III)-reduction. This is the first report of such an approach where the laborious procedures for protein purification are not required for identification of metal-reducing proteins. Such work provides the basis for a similar approach with other cultured organisms as well as analysis of sediment and groundwater samples from biostimulation efforts at contaminated sites. PMID- 17137662 TI - Heart rate response after emotional picture presentation is modulated by interoceptive awareness. AB - The perception of visceral signals plays a crucial role in many theories of emotions. The present study was designed to investigate the relationship between interoceptive awareness, emotional experience and heart rate responses in an emotional stimulation paradigm. Based on their performance in a heartbeat perception task 38 participants (16 males, 22 females) were classified as subjects with either high (n=19; 8 males) or low interoceptive awareness (n=19; 8 males). 120 pictures (40 pleasant, 40 unpleasant, 40 neutral slides) from the International Affective Picture System served as emotional stimuli. Heart rate changes were recorded during baseline and during slide presentation. After each slide, the subjects had to rate emotional valence and arousal on a 9-point self report scale. Statistical analyses revealed significantly stronger heart rate responses to pleasant and unpleasant stimuli in subjects with high interoceptive awareness. Furthermore, subjects with high interoceptive awareness rated pleasant and unpleasant slides as significantly more arousing; no differences were found in the emotional valence ratings. Heartbeat perception scores correlated significantly positive with both the mean arousal rating and with the mean heart rate changes. Our results demonstrate a strong relationship between the perception of cardiac signals and the peripheral processing of emotional stimuli. PMID- 17137663 TI - Effects of postural anxiety on the soleus H-reflex. AB - Previous research has proposed that spinal reflex modulation may mediate anxiety related changes in postural control. This study investigated how soleus H-reflex amplitude was influenced by standing at heights that induced different levels of anxiety. H-reflexes were elicited in 15 participants standing at the center and edge of a platform raised from a low to a high height (with and without vision). Increased skin conductance confirmed the anxiety effect of elevated surface heights. When standing at the edge of the platform with vision, H-reflex amplitude was attenuated in the high compared to low height condition. Changes in background muscle activity could not explain observed H-reflex changes, suggesting the potential involvement of pre-synaptic inhibition or fusimotor drive on anxiety-related changes in reflex modulation. This study reveals that healthy participants reduce spinal reflex excitability in the presence of increased postural anxiety and a postural threat imposed by standing at the edge of a raised platform. These findings have implications for understanding control of standing balance in individuals with postural instability and/or fear of falling, such as the elderly or stroke. PMID- 17137664 TI - Effects of novelty on activity of lateral and medial prefrontal neurons. AB - Detection of novel events is crucial for adapting to changing environments. The prefrontal cortex has been thought to be one of the areas involved in orienting attention to novel events. Here, we examined the effects of two components of novelty: context novelty, which purely happens when a familiar event occurs in an unpredicted situation or time and feature novelty, which happens by itself when an unfamiliar stimulus appears against the expectation of familiar ones. We trained monkeys on a task that included both novelty components and recorded the activity of neurons in the lateral and medial divisions of the prefrontal cortex. The responses of a substantial number of cells in both the lateral and medial divisions were enhanced when a familiar visual stimulus was presented in an unpredicted context. By contrast, enhancement of responses by the unfamiliarity of visual stimuli was observed mainly in cells in the lateral prefrontal cortex. These results suggest that the lateral and medial divisions of the prefrontal cortex are differentially involved in the control of attention triggered by novel sensory events. PMID- 17137665 TI - Constitutive activation of neuronal Src causes aberrant dendritic morphogenesis in mouse cerebellar Purkinje cells. AB - Src family tyrosine kinases are essential for neural development, but their in vivo functions remain elusive because of functional compensation among family members. To elucidate the roles of individual Src family members in vivo, we generated transgenic mice expressing the neuronal form of c-Src (n-Src), Fyn, and their constitutively active forms in cerebellar Purkinje cells using the L7 promoter. The expression of the constitutively active n-Src retarded the postnatal development of Purkinje cells and disrupted dendritic morphogenesis, whereas the wild-type n-Src had only moderate effects. Neither wild-type nor constitutively active Fyn over-expression significantly affected Purkinje-cell morphology. The aberrant Purkinje cells in n-Src transgenic mice retained multiple dendritic shafts extending in non-polarized directions and were located heterotopically in the molecular layer. Ultrastructural observation of the dendritic shafts revealed that the microtubules of n-Src transgenic mice were more densely and irregularly arranged, and had structural deformities. In primary culture, Purkinje cells from n-Src transgenic mice developed abnormally thick dendritic shafts and large growth-cone-like structures with poorly extended dendrites, which could be rescued by treatment with a selective inhibitor of Src family kinases, PP2. These results suggest that n-Src activity regulates the dendritic morphogenesis of Purkinje cells through affecting microtubule organization. PMID- 17137666 TI - Inflammatory bowel disease: current insights into pathogenesis and new therapeutic options; probiotics, prebiotics and synbiotics. AB - Inflammatory bowel disease (IBD) is a chronic relapsing disorder involving a dysregulated host-microbiota interaction. IBD patients have been shown to possess an increased risk for the development of colorectal cancer. Recently, focus has been placed on probiotic and prebiotic therapies, which aim to restore balance to the gastrointestinal microbiota, and reduce intestinal inflammation. Probiotics have been assessed extensively in animal models, with a number of clinical trials also demonstrating potential therapeutic benefits. However, it is widely accepted that more double-blind randomised placebo-controlled trials are required. Future research also needs to focus on determining which probiotics are the most efficacious in the IBD setting, and how the genetic and bacterial profiles of the patient will influence treatment responsiveness. Prebiotics have been studied less extensively, however, they may become an ideal treatment or co-treatment option due to their capacity to increase endogenous lactobacillus and bifidobacteria. Probiotics and prebiotics may offer a new therapeutic option for the treatment of IBD, however, a greater understanding of the mechanisms behind their action on the gastrointestinal microbiota is required in order to determine which probiotic, prebiotic or combinations thereof are the most beneficial. PMID- 17137667 TI - Stability and activity improvement of cephalosporin esterase EstB from Burkholderia gladioli by directed evolution and structural interpretation of muteins. AB - Esterase EstB from Burkholderia gladioli, which belongs to a family of esterases related to beta-lactamases and DD-peptidases was evolved for increased stability and simultaneously maintaining high cephalosporin C deacetylation activity. Random mutagenesis PCR was used to generate up to 5 aa substitutions per gene. A newly designed colony filter-screening assay, which was based on pH change after deacetylation of cephalosporin C in presence of DMF was established. In a first evolution round employing random mutagenesis, which included about 10(6) mutants, a set of interesting mutants was isolated. Distinct mutations identified as significant for stability were combined by a rational recombination step and the resulting recombinant was further evolved by an additional random mutagenesis round. After screening an additional 10(5) clones, it was possible to isolate a variant of EstB having more than 100-fold better activity in reactions containing 35% DMF. This mutant also showed a high increase in temperature stability (T(m) was raised by 13 degrees C) and retained high activity towards cephalosporin C under standard assay conditions. The molecular effects of mutations found in random mutants are discussed in view of the three-dimensional structure of wild type EstB. PMID- 17137668 TI - Towards understanding of the complex structure of growing yeast populations. AB - In a growing Saccharomyces cerevisiae population, cell size is finely modulated according to both the chronological and genealogical ages. This generates the complex heterogeneous structure typical of budding yeast populations. In recent years, there has been a growing interest in developing mathematical models capable of faithfully describing population dynamics at the single cell level. A multistaged morphologically structured model has been lately proposed based on the population balance theory. The model was able to describe the dynamics of the generation of a heterogeneous growing yeast population starting from a sub population of daughter unbudded cells. In this work, which aims at validating the model, the simulated experiment was performed by following the release of a homogeneous population of daughter unbudded cells. A biparametric flow cytometric approach allowed us to analyse the time course joint distribution of DNA and protein contents at the single cell level; this gave insights into the coupling between growth and cell cycle progression that generated the final population structure. The comparison between experimental and simulated size distributions revealed a strong agreement for some unexpected features as well. Therefore, the model can be considered as validated and extendable to more complex situations. PMID- 17137669 TI - Long acting local anesthetic-polymer formulation to prolong the effect of analgesia. AB - Prolonged postoperative analgesia cannot be achieved using single injections of local anesthetic solutions. The study objective was to evaluate the efficacy and toxicity of a new formulation of bupivacaine loaded in an injectable fatty acid based biodegradable polymer poly(sebacic-co-ricinoleic acid) for producing motor and sensory block when injected near the sciatic nerve. Bupivacaine was dissolved in poly(fatty ester-anhydride) paste and tested for drug release in vitro and in vivo after injection in mice. The efficacy and toxicity of the polymer-drug combination was determined by injecting the polymer formulation near the sciatic nerve of mice and measure the sensory and motor nerve blockade for 48 h, while monitoring the animal general health and the injection site. Seventy percent of the incorporated drug was released during 1 week in vitro. Single injection of 10% bupivacaine in the polymer caused motor and sensory block that lasted 30 h. Microscopic examination of the injection sites revealed only mild infiltration in three of eight examined tissues with no pathological findings for internal organs were found. In conclusion the polymer poly(sebacic-co-ricinoleic acid) is a safe carrier for prolonged activity of bupivacaine. PMID- 17137670 TI - Highly efficient and carcinoma-specific adenoviral replication restricted by the EGP-2 promoter. AB - Although some successes have been reported using adenoviral vectors for the treatment of cancer, adenoviral cancer gene therapy is still hampered by the lack of sufficient tumor cell killing. To increase the efficiency, adenoviruses have been modified to replicate specifically in tumor tissues by using tumor specific promoters controlling genes essential for adenoviral replication. However, many conditionally replicating adenoviral vectors replicate in one tumor type only, which limits their application. The epithelial glycoprotein-2 (EGP-2) promoter is active in a broad variety of carcinomas, the most common type of cancer. We utilized this promoter to restrict adenoviral replication. In this report we demonstrate that the potency of the replication-competent adenovirus AdEGP-2-E1 to specifically lyse EGP-2 positive cells is comparable to wild-type adenovirus (AdWT). In addition, we show that in vivo AdEGP-2-E1 replicates as efficient as AdWT in EGP-2 positive tumor cells. On the contrary, in EGP-2 negative cell lines as well as in primary human liver samples, the replication was attenuated up to 4 log in comparison to wild-type virus. This report clearly shows the potency of the EGP-2 promoter to mediate highly efficient and specific adenoviral replication for carcinoma gene therapy. PMID- 17137671 TI - [Multiple diabetic muscular infarctions after liver and kidney transplantation: a case report]. AB - INTRODUCTION: Several neuromuscular diseases may complicate diabetes mellitus and transplantation, including chronic sensorimotor length dependent polyneuropathy. OBJECTIVE: Description of muscular infarction, a rare complication of diabetes mellitus, which occurred after liver and kidney transplantation. CASE REPORT: A 57-year-old patient presented with long-term diabetes mellitus and multiple complications. End-stage renal and hepatic disease led to kidney and liver transplantation. Twenty-seven days after transplantation, swelling and induration appeared in the left shoulder and forearm. Forty-three days after transplantation, the same symptoms appeared in both lower limbs. Markedly reduced range of motion led to severe disability. Bone scans showed multiple spots following muscle anatomy. Computed tomography gave negative results. Magnetic resonance imaging (MRI) confirmed muscular infarction by a high T1 signal (muscular necrosis) and soft-tissue infiltration. DISCUSSION AND CONCLUSION: Muscular infarction is a rare and unknown complication of diabetes mellitus. It is characterised by sudden painful muscular induration and swelling affecting one muscle at a time with recurrence. Our patient presented with simultaneous multiple muscular infarctions in 3 limbs. Diagnosis was based on clinical investigation and MRI. The treatment is conservative and the condition generally resolved by itself. However, the long-term prognosis of muscular infarction is not good because of the cardiovascular-associated complications of diabetes mellitus. PMID- 17137672 TI - [Satisfaction with technological equipment in individuals with tetraplegia following spinal cord injury]. AB - OBJECTIVE: To measure the acquisition, use and satisfaction with high technology equipment by spinal cord injured tetraplegic subjects. DESIGN AND SETTING: A questionnaire was mailed to 102 tetraplegic subjects who were hospitalised in the rehabilitation center of Kerpape (Ploemeur, France) between 1998 and 2004, and 59 subjects responded. The questionnaire asked about the use of telephones, computers, wheelchairs and environmental controls at home. RESULTS: When a piece of equipment was acquired, it was very often used. Patient satisfaction with equipment was 79.3%. Home phones and mobile phones were often used with options such as hands-free devices (78 and 59% respectively). A total of 64.4% of subjects acquired a manual wheelchair and 61% a power wheelchair. The most commonly acquired options on the power wheelchairs were the powered recline (73,7%) and tilt (71,1%) systems. All options were used but all were more desired than acquired. A total of 27.1% of subjects desired a pushrim-activated power assist wheelchair, but only 15.3% had acquired one; 695% of subjects had a computer. Communication was the first use for the computer (82.5%); 49.2% of subjects had acquired an environmental control system, but 20% desired one. The first reason for lack of acquisition was financial difficulties but also accessibility and information problems. The factor that influenced the acquisition and need for equipment was the degree of spinal cord injury. No other factor reduced patient satisfaction with equipment. CONCLUSION: Patients were satisfied with the equipment they acquired. But their needs, especially wheelchair options and environmental control systems, were not satisfied. PMID- 17137673 TI - Patients with aortic valve stenosis and Type 2 diabetes have similar coronary flow reserve and aortic distensibility compared with non-diabetic patients with aortic valve stenosis. PMID- 17137674 TI - German physicians "on strike"--shedding light on the roots of physician dissatisfaction. AB - BACKGROUND: Over the past few years, students in Germany have been dropping out of medical school at increasing rates, and the number of physicians choosing to work abroad or in non-medical professions has been growing. A recent study (the "Ramboll Study") commissioned by the Health Ministry concluded that German physicians' dissatisfaction with existing monetary and non-monetary incentive systems during training and subsequent practice was the main reason for these trends. Among those physicians who have remained in the workforce, there is a similar dissatisfaction, reflected in part by a general strike in 2006 by German physicians in favour of higher wages and better working conditions. OBJECTIVE: To better understand the decision-making process of physicians which is highly determined by the satisfaction they experience in their work life and to extract the factors that contribute to their satisfaction. METHODS: We surveyed all physicians who spent more than 50% of their time in patient care (and less than 50% in research) at the teaching hospital of the Hannover Medical School (839, after exclusion of pre-test participants). Based on existing satisfaction studies, we designed a self-administered questionnaire that contained 28 items, including items measuring several dimensions of physician job satisfaction; the monetary and non-monetary incentives the physicians experienced in the recent past; other job-related potential confounding factors and socio-demographic questions. Respondents were asked to rate each job satisfaction item on five point Likert scales regarding both satisfaction with and importance of the item. Data were analysed using descriptive statistics, factor and correlation analyses. RESULTS AND DISCUSSION: Our data suggest that non-monetary factors are important determinants of physician job satisfaction, perhaps more important than monetary incentives that may augment or reduce physicians' base incomes. Factor analysis revealed seven principal factors of which decision-making and recognition, continuous education and job security, administrative tasks and collegial relationships were highly significant, specialized technology and patient contact were significant and research and teaching and international exchange were not significant in contributing to physician job satisfaction. CONCLUSION: This study sheds light to the underlying factors that contribute to physician job satisfaction in Germany, and it provides insights into the reasons for physicians leaving medical practice. In order for a health system to recruit and retain physicians, it may be necessary for a system's physician strategy to shift from focusing primarily on hard, monetary and compensation-related factors to a broader focus that incorporates the soft, non-monetary factors. The implementation of policies and management practices that reduce the time burden on physicians, and enhance physicians' participation in the development of patient care management processes and in managerial decisions that affect patient care appears to be crucial. The result will be increased job satisfaction among physicians, which is important to the future recruitment and retention of doctors, as well as to the productivity and quality of the services provided by this essential component of our medical care systems. PMID- 17137675 TI - Coevolution of symbiotic mutualists and parasites in a community context. AB - Recent advances in our knowledge of parasitic and mutualistic associations have confirmed the central role of coevolutionary interactions in population and community ecology. Here, we discuss the potential coevolutionary interdependence of the strength and specificity of symbiotic interactions with the complexity and productivity of their environment. We predict that interactions become less beneficial with increasing environmental quality and that the association of productivity with symbiont specificity depends on the relative strengths of tradeoffs between host range and other life-history parameters. However, as biotic complexity increases, pathogen specificity is predicted to decline, whereas mutualist specificity will increase. Testing these predictions on a geographical scale would contribute significantly to the predictive science of coevolution, and to our ability to manage biological interactions embedded in increasingly fragmented landscapes. PMID- 17137676 TI - Emphasizing the ecology in parasite community ecology. AB - In natural systems, individuals are often co-infected by many species of parasites. However, the significance of interactions between species and the processes that shape within-host parasite communities remain unclear. Studies of parasite community ecology are often descriptive, focusing on patterns of parasite abundance across host populations rather than on the mechanisms that underlie interactions within a host. These within-host interactions are crucial for determining the fitness and transmissibility of co-infecting parasite species. Here, we highlight how techniques from community ecology can be used to restructure the approaches used to study parasite communities. We discuss insights offered by this mechanistic approach that will be crucial for predicting the impact on wildlife and human health of disease control measures, climate change or novel parasite species introductions. PMID- 17137677 TI - Quantifying the physical and chemical mass transfer processes for the fate and transport of Co(II)EDTA in a partially-weathered limestone-shale saprolite. AB - The objective of the research is to quantify the relative contributions of physical and chemical mass transfer to the movement of Co(II/III)EDTA (chelates of Cobalt and Ethylene Diamine Tetraacetic Acid or EDTA) through a limestone shale saprolite soil. Saprolite is a collective term referring to partially weathered bedrock. It exists extensively in the subsurface. Because the parent bedding structures are maintained during the weathering process, saprolite soils are characterized by intensive fractures and secondary deposits of minerals such as Al-, Fe- and Mn-oxides on the fracture surfaces. Movement of reactive species through the soils may be influenced by diffusion into the rock matrix, a physical mass transfer (PMT) process, and interfacial chemical reactions, a chemical mass transfer (CMT) process. The PMT and CMT processes are phenomenologically similar but mechanistically different. In this research, previous laboratory observations from a Br and Co(II)EDTA tracer injection into an undisturbed saprolite soil column were used. Mechanistic reactive transport models were formulated to quantify the PMT and CMT processes. The PMT process was independently characterized by using the non-reactive tracer Br. Model parameters thus obtained were subsequently used as constraints to quantify the CMT processes involving Co(II)EDTA and its oxidation product Co(III)EDTA. Our calculations indicated that the PMT rates of the less reactive Co(III)EDTA were comparable with their theoretical CMT rates. In contrast, for the more reactive species Co(II)EDTA, CMT rates are higher than PMT rates. Evaluations of alternative CMT process models further confirmed one of our hypotheses on the basis of previous experimental understandings. The hypothesis suggested that competition from Fe-oxide for Co(II)EDTA may account for the majority of the decrease of Co(III)EDTA effluent concentrations that resulted in the separation of total Co and Co(III)EDTA breakthrough curves. Because Co(III)EDTA is more mobile than Co(II)EDTA in the subsurface, the results of this research suggest independent quantifications of CoEDTA PMT and CMT processes if laboratory results are to be interpreted correctly and scaled up for field and predictive uses. PMID- 17137678 TI - You eat what you are: modern health worries and the acceptance of natural and synthetic additives in functional foods. AB - There is an increasing array of functional foods available that are designed to confer health benefits. However, individuals' worries about new technology and modernity may influence the acceptance of these products. In this study, we investigated how modern health worries influence attitudes and decisions about functional foods. We asked participants (n=390) to rate pictures of products with either added vitamins or added scientific compounds. Each product shown purported to have one of three possible targeted effects: to reduce the likelihood of a disease, to reduce a risk factor associated with a disease, or to improve personal appearance. We found levels of modern health worries to be significantly associated with participants' reports of organic food consumption and presence of food allergies. Modern health worries were also significantly related to a preference for foods with natural as opposed to synthetic additives. Participants with higher levels of modern health worries had a greater acceptance of functional foods designed to reduce the likelihood of disease compared to participants with low modern health worries. Overall, the results suggested that modern health worries are an important psychological factor to consider with regards to attitudes toward functional foods. PMID- 17137679 TI - Synthesis and anticancer activity of a novel class of flavonoids: 2,4 diarylchromane[4,3-d]-delta(1,9b)-1,2,3-thiadiazolines. AB - A series of 2,4-diarylchromane[4,3-d]-Delta(1,9b)-1,2,3-thiadiazolines have been synthesized by cyclization of corresponding 2-arylchroman-4-one-arylhydrazones with SOCl(2) then treated with alcohol. All the compounds have been tested for their antiproliferative activity in vitro against six human tumor cell lines, and the highly potent derivative 11a exhibited in vivo inhibitory effect on tumor growth. Mechanism research indicated that it is due to 11a that induces DNA fragmentation. PMID- 17137680 TI - [Results and place of Fibroscan in the non-invasive diagnosis of hepatic fibrosis]. AB - BACKGROUND AND AIMS: Diagnosis of liver fibrosis by transient elastography (Fibroscan) is a new and non-invasive method. We report its results, and compare them with the non-invasive biological tests. RESULTS: Fibroscan is validated for chronic hepatitis C, allowing to diagnose the fibrosis (Metavir) F2, F3 and F4 with areas under the ROC curve of 0.79-0.83, 0.90-0.91 and 0.91-0.97 respectively. The non-invasive diagnosis of the cirrhosis can be made with positive and negative predictive values from 70 to 95% and 77 in 95%. Preliminary data concern the HCV-HIV co-infections, primary biliary cirrhosis, NASH, portal hypertension, and the follow-up of the hepatic fibrosis. These results are altogether comparable to the various non-invasive biological tests, for cirrhosis diagnosis. CONCLUSIONS: It's remain to determine the respective places of Fibroscan and non-invasive biological tests, in the liver fibrosis diagnosis. PMID- 17137681 TI - [Multi-organ failures during septic shock from Escherichia coli urinary tract infection: catastrophic antiphospholipid syndrome?]. AB - INTRODUCTION: Although the presence of antiphospholipid antibodies is not an uncommon fact during infection, their responsibility in serious manifestations can still be debated and questions the existence of an actual catastrophic antiphospholipid syndrome or multivisceral faintings, triggered by the infection only, since the presence of antiphospholipid antibodies is not pathogenous. CASE: A 68-year-old man presented during an Escherichia coli urinary tract infection a septic shock with renal and cardiac insufficiencies, hepatic cytolysis and cholestasis and disseminated intravascular coagulation. There was a significant titer of antiphospholipid antibodies IgG (50 UGPL/ml) with an antibeta2 glycoprotein-1 positivity. The patient quickly recovered with antibiotherapy and intravenous immunoglobulins. DISCUSSION: Diagnosing the pathogeny of multivisceral faintings is founded of the clinical manifestations published during the catastrophic antiphospholipid syndrome, the evolution and the persistence of post recovery antibodies and the comparison with the visceral bouts that the sepsis exclusively as revealed. PMID- 17137682 TI - [HIV infection and spontaneous control of the infection]. PMID- 17137683 TI - [Update in Hepatitis C virus associated extrahepatic manifestations]. AB - INTRODUCTION: Since the discovery of the hepatitis C virus, many manifestations, so called extra-hepatic manifestations (EHM), are largely reported with more or less relationship proofs. ACTUALITIES AND MAIN POINTS: This article proposes a review of the main extra-hepatic manifestations associated with the Hepatis C Virus infection and which remain a topical subject, more than fifteen years after the discovery of this virus. Mixed cryoglobulin and its vasculitic manifestations are still one of the more frequent Hepatis C Virus associated-extra-hepatic manifestations. Its management may be critically changed due to the increasing use of anti-CD20 therapy. Among other HCV-EHM, the following extra-hepatic manifestations are still of interest: the chronic fatigue syndrome, the sicca syndrome, the non-insulin-dependent diabetes mellitus, malignant B cell proliferations, mainly the Hepatis C Virus-related splenic lymphoma with villous lymphocytes and the production of auto-antibodies. PERSPECTIVES AND PROJECTS: The mechanisms underlying these HCV-associated EHM are ill-elucidated and still remain of great interest as proved by current studies. The use of anti-CD20 antibodies in the treatment of cryoglobulinemic vasculitis is also under investigation. PMID- 17137684 TI - Independent extended supplementary nurse prescribers, their prescribing practice and confidence to educate and assess prescribing students. AB - BACKGROUND: No other country in the world has such extended prescribing rights for nurses as the United Kingdom. Concerns surround the move of nursing towards a medical model of care, and the level of medical practice support required by trainee prescribers. AIM: To provide an overview of the nurses adopting the role of independent extended supplementary prescriber, their prescribing practice and confidence to educate and assess prescribing students. METHODS: A convenience sample of 1187 independent extended supplementary nurse prescribers were sent a questionnaire. Eight hundred and sixty eight completed questionnaires were returned. RESULTS: The majority (82%) of nurses worked in primary care. Eighty seven percent used independent extended prescribing and 35% supplementary prescribing. Most were qualified to degree level or higher and had over 10 years nursing experience. Seventy four percent felt confident to act as a mentor during the prescribing programme. More highly qualified nurses and those who had undertaken, or had access to continuing professional development, were statistically more likely to feel confident to adopt this role. CONCLUSION: Appropriately qualified nurse prescribers might be best placed to support trainee prescribers. Exploration of the low uptake of supplementary prescribing and access to continuing professional development is required. PMID- 17137685 TI - Variation in transfer factor of radiocaesium in bank voles (Clethrionomys glareolus) in clear cut and mature forest sites after the Chernobyl accident. AB - Bank voles that were collected between 1986 and 2004 at sites in Chernobyl fallout areas of northern Sweden showed higher (137)Cs activity concentrations at the mature forest sites compared to clear cuts. This difference was not attributed to differences in ground deposition between sites but to differences in aggregated transfer rates to voles. Differences in transfer between forest types were evident for all years 1986-2004 but the change occurred at different rates in the two habitats. The apparent transfer factor between bilberry (Vaccinium myrtillus) and voles was positively related and indicated that a biomagnification was about 1.5 from vegetation to these small mammalian herbivores. The aggregated transfer factor to bank voles measured in the forest habitat, although starting at higher levels declined faster with time than clear cut sites and the differences between the forest habitat and the clear cut areas diminished with time. After the Chernobyl accident in 1986 the mean level in bank vole was 514Bq/kg fresh mass (SD=505) that increased to 1485Bq/kg (SD=881) in 1988. The activity concentration declined thereafter. The bank voles collected in similar habitats in 2004 contained on average 1022Bq/kg (SD=723). Still 18 years after the radionuclide fallout over Sweden high activity concentrations in voles could be found. PMID- 17137686 TI - WITHDRAWN: Stereolithography and the use of pre-adapted or fabricated plates for accurate repair of maxillofacial defects. AB - This article has been withdrawn consistent with Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). The Publisher apologizes for any inconvenience this may cause. PMID- 17137687 TI - Re: Peripheral nerve compression neuropathy after fractures of the distal radius. PMID- 17137688 TI - Chronic osteomyelitis of the carpal trapezoid. PMID- 17137689 TI - Validation of predicted exponential concentration profiles of chemicals in soils. AB - Multimedia mass balance models assume well-mixed homogeneous compartments. Particularly for soils, this does not correspond to reality, which results in potentially large uncertainties in estimates of transport fluxes from soils. A theoretically expected exponential decrease model of chemical concentrations with depth has been proposed, but hardly tested against empirical data. In this paper, we explored the correspondence between theoretically predicted soil concentration profiles and 84 field measured profiles. In most cases, chemical concentrations in soils appear to decline exponentially with depth, and values for the chemical specific soil penetration depth (d(p)) are predicted within one order of magnitude. Over all, the reliability of multimedia models will improve when they account for depth-dependent soil concentrations, so we recommend to take into account the described theoretical exponential decrease model of chemical concentrations with depth in chemical fate studies. In this model the d(p)-values should estimated be either based on local conditions or on a fixed d(p)-value, which we recommend to be 10cm for chemicals with a log K(ow)>3. PMID- 17137690 TI - Toxicological responses of red-backed salamanders (Plethodon cinereus) to subchronic soil exposures of 2,4-dinitrotoluene. AB - Dinitrotoluenes are used as propellants and in explosives by the military and as such have been found at relatively high concentrations in the soil. To determine whether concentrations of 2,4-dinitrotoluene (2,4-DNT) in soil are toxic to amphibians, 100 red-backed salamanders (Plethodon cinereus) were exposed to either 1500, 800, 200, 75 or 0mg 2,4-DNT/kg soil for 28 days and evaluated for indicators of toxicity. Concentrations of 2,4-DNT were less than targets and varied with time. Most salamanders exposed to concentrations exceeding 1050 mg/kg died or were moribund within the first week. Salamanders exposed to soil concentrations exceeding 345 mg/kg lost >6% of their body mass though no mortality occurred. Overt effects included a reduction in feed consumption and an increase in bucco-pharyngeal oscillations in salamanders. These results suggest that only high soil concentrations of 2,4-DNT have the potential to cause overtly toxic effects in terrestrial salamanders. PMID- 17137691 TI - The leachability, bioaccessibility, and speciation of Cu in the sediment of channel catfish ponds. AB - There have been growing concerns about the environmental impact of Cu applied in the catfish pond aquaculture. In this paper, sediments taken from three commercial catfish ponds were studied for content, leachability, bioaccessibility, and speciation of sediment-bound Cu. Results showed that the Cu was concentrated in the top 10 cm of the sediments and the peak Cu concentrations ranged from the background level to about 200 mg/kg. Toxicity characteristic leaching procedure showed only 1-8% of sediment Cu was leachable while bioaccessible Cu, evaluated by physiological based extraction test, accounted for up to 40-85% of total Cu. Due to the high redox potential in the surface sediments, acid-volatile sulfide was not a significant binding phase. The sequential extraction results showed that the residual phase (forms in lattices of primary and secondary minerals) was the major Cu fraction in the first two pond sediments but carbonate-bound, Fe/Mn oxide-bound and organically bound Cu, as well as the residual fraction, seemed equally important in the third pond. PMID- 17137692 TI - Drought changes the dynamics of trace element accumulation in a Mediterranean Quercus ilex forest. AB - We conducted a field drought manipulation experiment in an evergreen oak Mediterranean forest from 1999 to 2005 to investigate the effects of the increased drought predicted for the next decades on the accumulation of trace elements that can be toxic for animals, in stand biomass, litter and soil. Drought increased concentrations of As, Cd, Ni, Pb and Cr in roots of the dominant tree species, Quercus ilex, and leaf Cd concentrations in Arbutus unedo and of Phillyrea latifolia codominant shrubs. The increased concentration of As and Cd can aggravate the toxic capacity of those two elements, which are already next or within the levels that have been shown to be toxic for herbivores. The study also showed a great reduction in Pb biomass content (100-135 gha(-1)) during the studied period (1999-2005) showing the effectiveness of the law that prohibited leaded fuel after 2001. The results also indicate that drought increases the exportation of some trace elements to continental waters. PMID- 17137693 TI - Managing risk: risk perception, trust and control in a Primary Care Partnership. AB - In this article, managers' perceptions of risk on entering a newly formed primary health care partnership are explored, as are the mechanisms of trust and control used to manage them. The article reports a qualitative component of a 2-year National Health and Medical Research Council funded study of trust within the structures of a Primary Care Partnership (PCP) in Victoria, Australia. Multiple methods of data collection were employed. We found that managers identified risks at system, partnership and agency levels, and that as trust was built, concern about risks diminished. Trust effectively facilitated joint action, but it was betrayed on occasions, in which case the informal power of group process was used to contain the problems. The implications of this study for policy makers are in terms of understanding how perceptions of risk are constructed, the ways managers use social control to create a safer context in which to locate the trust-based relationships that facilitate joint action, and the importance of institutional arrangements. Without trust, joint action is hard to achieve, and without control, it is difficult to prevent breaches of trust from inhibiting joint action. PMID- 17137694 TI - HIV/AIDS and the construction of Sub-Saharan Africa: heuristic lessons from the social sciences for policy. AB - There is no doubt that Sub-Saharan African countries face major problems due to the HIV/AIDS pandemic that has ravaged the region. Yet the Eurocentric construction of the region as the source of the virus not only creates negative stereotypes in social science disciplines like geography, but also glosses over the potential of social science disciplines to provide knowledge and influence policy about HIV/AIDS. This oppositional construction of the region has unfortunately contributed to a glossing over of many aspects of Sub-Saharan Africa's people, their environment, culture, history, politics, economics, gender relations, and the region's global status that would provide important input for policy aimed at curbing the devastating spread of HIV in the region. This paper argues that once we recognize that HIV is a global virus with trans-cultural implications, social science disciplines, such as geography, can reveal certain attributes about the region and its HIV/AIDS pandemic that can be used in policy formulation to combat the spread of the virus. PMID- 17137695 TI - The social distribution of risk at work: acute injuries and physical assaults among healthcare workers working in a long-term care facility. AB - The roles of informal social ties in affecting healthcare workers' risk of injury and assault were investigated in a long-term care facility for the elderly in the US. The original hypothesis was that nurses and healthcare assistants who integrated more with their coworkers would have lower risk. A crude measure of familiarity and social integration with coworkers was constructed from staff attendance records. This variable, which indicates working a floor and shift one has routinely worked on in the past, was associated with a moderate increase in risk of being injured after controlling for lifting demands and a fairly strong increased risk of being assaulted after controlling for resident combativeness. An interaction between social integration and job title was found. The primary associations were in the opposite direction of what was expected. The results suggest that social forces among healthcare workers shape the distribution of risk among workers in a manner more complex than some theories of social integration have suggested. New hypotheses are proposed to explore how social norms and expectations affect the way workers interact with each other and shape the distribution of risk among workgroup members. PMID- 17137696 TI - [Early preventive pressure therapy in extensive skin grafts on children's face, a comeback of classic methods? A retrospective study on 8 cases]. AB - INTRODUCTION: One of the common techniques in the treatment of giant facial naevi is the excision covered by a skin graft. It's functional and esthetic results are not always excellent. Physiotherapy including immediate postsurgical preventive pressure therapy could ameliorate these results and shorten the period of scarring. PURPOSE: Evaluation of scarring results with early pressure therapy in skin grafts of congenital facial naevi. PATIENTS AND METHODS: Retrospective study on 8 children being 1 to 9 years old and presenting congenital facial naevi. All of them were treated by excision followed by full thickness skin grafting. The associated physiotherapy was realized by a transparent rigid or elastic mask. The pressure was started after skin graft uncovering between 7 and 20 days. The period for this treatment was depending on the maturing of the scar. Evaluation was established by 3 examinators on: time till scar maturing, trame quality and esthetic quality. RESULTS: The period till definitive scar maturing was showed to be between 8 and 14 month; trame quality was mostly evaluated as excellent, the esthetic result oscillated between good and medium. CONCLUSION: Physiotherapy seems to be an essential association for the treatment of facial full thickness skin grafts in children. Early pressure therapy could better the functional and esthetic result and accelerate scar maturing. Simple skin grafting could become the first treating option in this indication. Future prospective studies are expected to confirm these results. PMID- 17137697 TI - [Modified Texier mediodorsal transposition island skin flap of the nose. Report of four cases of ala nasi reconstruction]. AB - First described by Texier in 1994, the mediodorsal transposition flap of the nose is an island flap. The paramedian dorsal arteries (anastomosed to the interdomal plexus) ensured the axial vascularisation in SMAS plane. The authors will discuss their personal approach to the surgical procedure for the improvement of final result, with four clinical cases. This flap can be used for aesthetic unit reconstruction of cutaneous or mucosal different alar defect (partial or complete, full-thickness or not). This reliable flap represents an alternative technique of composed grafts, of different nasolabial flaps, and of forehead flap. PMID- 17137698 TI - [Difficulties of the management of head and neck neurofibromatosis]. AB - INTRODUCTION: Neurofibromatoses represent a group of 7 diseases having the same cutaneous signs due to a common embryologic origin. The types 1 and 2 are the most frequent and the most studied. Neurofibromatosis type 1 (NF1) or Von Recklinghausen's neurofibromatosis is an autosomal dominantly inherited disease, whose prevalence is 1/4500. The cranio-orbitotemporal lesions exist in 1 to 10% of the cases. Neurofibromatosis type 2 (NF2) is also an autosomal dominantly inherited disease, but is ten times less frequent than the NF1 and is characterized by bilateral vestibular schwannomas (former acoustic neurinomas). The purpose of this work was to detail the clinical features, highlighting the diagnostic and therapeutic difficulties. MATERIAL AND METHODS: Nine patients managed between 1990 and 2005 were reviewed retrospectively. Eight patients were found carrier of NF1 and one patient carrier of NF2 according to the diagnostic criteria of the 1988's National Institute of Health consensus. RESULTS: Five women and four men were managed; patient age varied between 13 and 62 years and the mean age was 26 years. Surgical resection of the tumor was carried out among 8 patients with bone reconstruction of the orbit on 3 patients and cutaneous expansion on one other. The results are difficult to appreciate especially that the surgery was iterative. However, all our patients were satisfied with the result and we noted recurrence in a patient presenting a retro-auricular tumor, which filled the external auditory canal requiring a further surgery. The patient having NF2 was treated by neurosurgery and showed a good result. DISCUSSION: The NF1 manifestations are extremely variable. Facial neurofibromatoses are difficult to control given the presence of soft tissues infiltration and the associated osseous dysplasia. The results are modest and recurrence is frequent. Cancer risks and the disease's completely unpredictable evolution urge a regular and multidisciplinary patient follow-up. PMID- 17137699 TI - [The total thigh flap in reconstructive surgery]. AB - Plastic surgeons have sometimes to face large pelvic tissue defects, particularly in paraplegic patients with recurrent pressure sores. This is frequently a reconstructive challenge and different locoregional flaps may be useful to manage those wounds when medical solutions were unsuccessful. However, in cases when other reconstructive options have been exhausted the total thigh flap represents the last acceptable solution to cover the defects by providing a large volume of healthy tissue. We remind here the operative technique and its interest in reconstructive surgery by reporting our experience in two patients for which this flap was successfully used. PMID- 17137700 TI - [Breast implants from 1957 changed on 2005: a 48-year exceptional and historical follow-up]. AB - A 78-year-old woman underwent a breast implants removal on August 2005. The prosthetic breast augmentation was performed during 1957, in France. The photos of these polyethylene made breast implants are shown. PMID- 17137701 TI - The South-East Asian tsunami disaster--how Resuscitation helped the recovery programme. PMID- 17137702 TI - Decreased fluid volume to reduce organ damage: a new approach to burn shock resuscitation? A preliminary study. AB - OBJECTIVE: To evaluate the impact of decreased fluid resuscitation on multiple organ dysfunction after severe burns. This approach was referred to as "permissive hypovolaemia". METHODS: Two cohorts of patients with burns>20% BSA without associated injuries and admitted to ICU within 6 h from the thermal injury were compared. Patients were matched for both age and burn severity. The multiple-organ dysfunction score (MODS) by Marshall was calculated for 10 days after ICU admission. Permissive hypovolaemia was administered by a haemodynamic oriented approach throughout the first 24-h period. Haemodynamic variables, arterial blood lactates and net fluid balance were obtained throughout the first 48 h. RESULTS: Twenty-four patients were enrolled: twelve of them received the Parkland Formula while twelve were resuscitated according to the permissive hypovolaemic approach. Permissive hypovolaemia allowed for less volume infusion (3.2+/-0.7 ml/kg/% burn versus 4.6+/-0.3 ml/kg/% burn; P<0.001), a reduced positive fluid balance (+7.5+/-5.4 l/day versus +12+/-4.7 l/day; P<0.05) and significantly lesser MODS Score values (P=0.003) than the Parkland Formula. Both haemodynamic variables and arterial blood lactate levels were comparable between the patient cohorts throughout the resuscitation period. CONCLUSIONS: Permissive hypovolaemia seems safe and well tolerated by burn patients. Moreover, it seems effective in reducing multiple-organ dysfunction as induced by oedema fluid accumulation and inadequate O2 tissue utilization. PMID- 17137703 TI - Leptin gene expression in subcutaneous adipose tissue in girls before and during puberty. AB - OBJECTIVES: Leptin functions as a neuroendocrine hormone and it is related to the onset of puberty in animal models. Its role in normal human sexual maturation is still incompletely defined. The aim of the study was to assess the relationships between leptin mRNA (gene) expression, thickness of subcutaneous fat tissue and the serum concentration of leptin in girls before and during puberty. STUDY DESIGN: Twenty-nine lean girls were studied (mean age 10.8+/-1.9 years). The subjects were divided into two groups according to pubertal status. The first group consisted of 14 prepubertal girls and second group of 15 girls who were in puberty. Body height, weight, arm circumference, skin fold thickness at abdominal, triceps and subscapular sites were measured. Serum leptin was assessed by RIA method. Leptin mRNA was measured in subcutaneous abdominal adipose tissue by semi-quantitative assays based on reverse transcription (RT) of the mRNA and polymerase chain reaction (PCR) amplification of the cDNA. RESULTS: Girls in pubertal stages had higher serum leptin concentration than prepubertal girls. The mean values of leptin mRNA level in subcutaneous abdominal adipose tissue were not statistically different between groups. There was also no difference between the thickness of skin folds in investigated girls. A positive correlation between leptin mRNA expression and skin fold thickness, BMI and arm circumference as well as between the leptin concentration and skin fold thickness, BMI and arm circumference were observed. CONCLUSIONS: The level of leptin gene expression and serum leptin concentrations depend on the amount of fat tissue. We can propose that initiation of pubertal events does not result from increased of leptin mRNA expression in subcutaneous abdominal fat cells or from its increased concentration in blood. PMID- 17137704 TI - A fertility-preserving option in early cervical carcinoma: laparoscopy-assisted vaginal radical trachelectomy and pelvic lymphadenectomy. AB - OBJECTIVE: Laparoscopic vaginal radical trachelectomy is a fertility-preserving alternative to radical hysterectomy or chemoradiation in young women with stage IA2 to IB cervical cancers. The aim of this study is to describe the feasibility and outcome of laparoscopic radical vaginal trachelectomy and pelvic lymphadenectomy in women treated with early cervical cancers who wish to maintain fertility. STUDY DESIGN: From August 2000 through December 2004, 16 patients were offered this procedure. Patients were selected for this treatment on the basis of favorable cervical tumors and a desire to maintain fertility. Laparoscopic pelvic lymphadenectomies were performed in all patients. Vaginal radical trachelectomy was performed immediately if removed lymph nodes were negative. Obstetrical and oncological outcomes were evaluated. RESULTS: Sixteen women underwent this procedure. The mean operative time was 142 min (115-178 min), with a mean blood loss of 180 ml (120-230 ml), and the average hospital stay was 6.7 days. No intraoperative or postoperative complications occurred. With an average follow-up of 28.2 months, there have been no recurrences. Five pregnancies have subsequently occurred, with two third-trimester deliveries, two miscarriages at 24 and 26 weeks' gestation, and one patient is currently 18 weeks' pregnant. CONCLUSIONS: Laparoscopy-assisted radical vaginal trachelectomy is an adequate treatment, with its minimally invasive procedure and shorter recovery time, for early-stage cervical cancer in women who wish to preserve fertility. However, fertility issues remain the largest unanswered problem with this technique. PMID- 17137705 TI - Interactions of chlorpromazine with phospholipid monolayers: effects of the ionization state of the drug. AB - Molecular dynamics simulations have been performed to investigate the interactions between chlorpromazine (CPZ) and Langmuir monolayers of the zwitterionic dipalmitoylphosphatidylcholine (DPPC) and the anionic dipalmitoylphosphatidylglycerol (DPPG). Simulations for a fixed surface density and different charge states - neutral and protonated CPZ - were able to capture important features of the CPZ-phospholipid monolayer interaction. Neutral CPZ is predominantly found in the hydrophobic tail region, whereas protonated CPZ is located at the lipid-water interface. Specific interactions (hydrogen bonds) between protonated CPZ and the lipid head groups were found for both zwitterionic and anionic monolayers. We computed lipid tail order parameters and investigated the effects of the drug upon tail ordering. We also computed electrostatic surface potentials and found qualitative good agreement with experimental results. PMID- 17137706 TI - Carbon sinks and sources in China's forests during 1901-2001. AB - This paper reports the annual carbon (C) balance of China's forests during 1901 2001 estimated using the Integrated Terrestrial Ecosystem C-budget model (InTEC). Annual carbon source and sink distributions are simulated for the same period using various spatial datasets including land cover and leaf area index (LAI) obtained from remote sensing, soil texture, climate, forest age, and nitrogen deposition. During 1901-1949, China's forests were a source of 21.0+/-7.8 Tg C yr(-1) due to disturbances (human activities). Its size increased to 122.3+/-25.3 Tg C yr(-1) during 1950-1987 due to intensified human activities in the late 1950s, early 1960s, 1970s and early 1980s. The forests became large sinks of 176.7+/-44.8 Tg C yr(-1) during 1988-2001, owing to large-scale plantation and forest regrowth in previously disturbed areas as well as growth stimulation by nondisturbance factors such as climatic warming, atmospheric CO(2) fertilization, and N deposition. From 1901 to 2001, China's forests were a small carbon source of 3.32 Pg C, about 32.9+/-22.3 Tg C yr(-1). The overall C balance in biomass from InTEC generally agrees with previous results derived from forest inventories of China's forests. InTEC results also include C stock variation in soils and are therefore more comprehensive than previous results. The uncertainty in InTEC results is still large, but it can be reduced if a detailed forest age map becomes available. PMID- 17137707 TI - Assessing the predictive validity and efficacy of a multimodal training programme for laparoscopic radical prostatectomy (LRP). AB - OBJECTIVES: To assess the predictive validity (ability to correlate to real-life environment) and efficacy of a training programme for laparoscopic radical prostatectomy (LRP), based on a structured and progressive pelvitrainer component with hands-on clinical training in the operating room (OR). METHODS: Prospective data on 500 LRP cases were analysed with 80 excluded due to incomplete records. The operation was divided into multiple steps. Times for these steps were compared among 11 surgeons with different laparoscopic expertise (first-, second , and third-generation surgeons in order of decreasing experience) and correlated to times for specific exercises on the pelvitrainer that simulated particular steps. Perioperative parameters were also evaluated among the three groups. RESULTS: Pelvitrainer times achieved by trainees (third-generation surgeons) did not differ significantly with times for corresponding steps of LRP. There was also no significant difference for total OR time between the second- and third generation surgeons (205 and 207 min, respectively; p>0.05) although the time for the first-generation surgeons was faster than both (176 min). Short-term quality indicators for first, second, and third generations included transfusion rates (2.3%, 2.4%, and 2.6%, respectively), positive margin rates (20.3%, 21.5%, and 23.0%) and complications, which did not differ significantly among the generations although the first-generation surgeons had the lowest rates. CONCLUSIONS: A carefully designed training programme that incorporates both pelvitrainer and mentor-based operative training is essential for the effective and safe transfer of skills and knowledge required to learn LRP. PMID- 17137708 TI - Anatomical landmarks of radical prostatecomy. AB - INTRODUCTION: In the present study, we review current literature and based on our experience, we present the anatomical landmarks of open and laparoscopic/endoscopic radical prostatectomy. METHODS: A thorough literature search was performed with the Medline database on the anatomy and the nomenclature of the structures surrounding the prostate gland. The correct handling of puboprostatic ligaments, external urethral sphincter, prostatic fascias and neurovascular bundle is necessary for avoiding malfunction of the urogenital system after radical prostatectomy. RESULTS: When evaluating new prostatectomy techniques, we should always take into account both clinical and final oncological outcomes. The present review adds further knowledge to the existing "postprostatectomy anatomical hazard" debate. It emphasizes upon the role of the puboprostatic ligaments and the course of the external urethral sphincter for urinary continence. When performing an intrafascial nerve sparing prostatectomy most urologists tend to approach as close to the prostatic capsula as possible, even though there is no concurrence regarding the nomenclature of the surrounding fascias and the course of the actual neurovascular bundles. After completion of an intrafascial technique the specimen does not contain any periprostatic tissue and thus the detection of pT3a disease is not feasible. This especially becomes problematic if the tumour reaches the resection margin. DISCUSSION: Nerve sparing open and laparoscopic radical prostatectomy should aim in maintaining sexual function, recuperating early continence after surgery, without hindering the final oncological outcome to the procedure. Despite the different approaches for radical prostatectomy the key for better results is the understanding of the anatomy of the bladder neck and the urethra. PMID- 17137709 TI - Placenta growth factor not vascular endothelial growth factor A or C can predict the early recurrence after radical resection of hepatocellular carcinoma. AB - The purpose of this study was to evaluate the relationship between the expression of PlGF in tumor tissue and clinical outcomes in HCC patients. Tumor PlGF and vascular endothelial growth factor (VEGF)-A and VEGF-C mRNA were analyzed. Results demonstrated that patients with PlGF expression levels higher than median tended to have early recurrence compared to patients with PlGF expression lower than median (P=.031). In patients with AJCC stage II-III disease, this difference was even more significant (P=.002). In contrast, VEGF-A and VEGF-C could not predict early recurrence-free survival. Since PlGF expression correlated with early recurrence of HCC, PlGF may be an important prognostic indicator in HCC. PMID- 17137710 TI - Avemar, a nontoxic fermented wheat germ extract, induces apoptosis and inhibits ribonucleotide reductase in human HL-60 promyelocytic leukemia cells. AB - Avemar (MSC) is a nontoxic fermented wheat germ extract demonstrated to significantly improve the survival rate in patients suffering from various malignancies. We investigated its effects in human HL-60 promyelocytic leukemia cells. After 24, 48, and 72 h of incubation, Avemar inhibited the growth of HL-60 cells with IC50 values of 400, 190, and 160 microg/ml, respectively. Incubation with MSC caused dose-dependent induction of apoptosis in up to 85% of tumor cells. In addition, Avemar attenuated the progression from G2-M to G0-G1 phase of the cell cycle and was also found to significantly reduce the in situ activity of ribonucleotide reductase, the key enzyme of de novo DNA synthesis. We conclude that Avemar exerts a number of beneficial effects which could support conventional chemotherapy of human malignancies. PMID- 17137711 TI - Optimising photoelectrocatalytic oxidation of fulvic acid using response surface methodology. AB - In this paper, statistics-based experimental design with response surface methodology (RSM) was employed to investigate the effect of operation conditions on photoelectrocatalytic oxidation of fulvic acid (FA) using a Ti/TiO(2) electrode in a photoreactor. Initially, the Box-Behnken design was employed including the three key variables (initial pH, potassium peroxodisulphate (K(2)S(2)O(8)) and bias potential). Thereafter, the mutual interaction and effects between these parameters and optimum conditions were obtained in greater detail by means of SAS and Matlab software. The results of this investigation reveal that: (1) the regression analysis with R(2) value of 0.9754 shows a close fit between the experimental results and the model predictions; (2) three dimension response surface plot can provide a good manner for visualizing the parameter interactions; (3) the optimum pH, K(2)S(2)O(8) and bias potential is found to be 3.8, 88.40 mg/L, 0.88 V, respectively, and the highest FA removal efficiency of 57.06% can be achieved. PMID- 17137712 TI - Dye wastewater treated by Fenton process with ferrous ions electrolytically generated from iron-containing sludge. AB - Fenton process was employed to treat synthetic dye wastewater with supply of Fe(II) electrolytically generated from iron-containing sludge which was recycled and reused throughout the study. Treated water quality and properties of iron sludge after being repeatedly used were reported and discussed. Experimental results showed that COD was mainly removed by oxidation other than coagulation. Although, the process was quite effective for COD and color removal, conductivity of treated water was enormously high. Meanwhile, repeated use of iron-containing sludge results in accumulation of organic materials embedded in the sludge as indicated by increasing volatile suspended solid (VSS)/TSS ratio and decreasing zeta potential. PMID- 17137713 TI - Potential for BLEVE associated with marine LNG vessel fires. AB - Recent LNG marine shipping hazard studies have discounted BLEVE hazards associated with LNG vessels. This exclusion of a potential major hazard event has been queried, particularly since a recent LNG truck BLEVE-like event in Spain. This paper reviews the physical factors associated with the Spanish LNG truck event and accepts that this had features of a classical BLEVE event and that there is no inherent property of LNG excluding BLEVE-like events, although US LNG trucks would be safer due to design features. Marine LNG vessels have differently designed tanks and it is demonstrated that the combination of physical barriers makes direct thermal input to the LNG inner tank more limited than hypothesized by some, but if it occurs these tanks cannot rise to a pressure sufficient to cause a large flash of liquid and consequent BLEVE event of a scale hypothesized in the literature. PMID- 17137714 TI - Differential brain opioid receptor availability in central and peripheral neuropathic pain. AB - This study used positron emission tomography (PET) and [11C]diprenorphine to compare the in vivo distribution abnormalities of brain opioid receptors (OR) in patients with peripheral (n=7) and central post-stroke pain (CPSP, n=8), matched for intensity and duration. Compared with age- and sex-matched controls, peripheral neuropathic pain (NP) patients showed bilateral and symmetrical OR binding decrease, while in CPSP binding decrease predominated in the hemisphere contralateral to pain. In CPSP patients, interhemispheric comparison demonstrated a significant decrease in opioid binding in posterior midbrain, medial thalamus and the insular, temporal and prefrontal cortices contralateral to the painful side. Peripheral NP patients did not show any lateralised decrease in opioid binding. Direct comparison between the central and peripheral groups confirmed a significant OR decrease in CPSP, contralateral to pain. While bilateral binding decrease in both NP groups may reflect endogenous opioid release secondary to chronic pain, the more important and lateralised decrease specific to CPSP suggests opioid receptor loss or inactivation in receptor-bearing neurons. Opioid binding decrease was much more extensive than brain anatomical lesions, and was not co-localised with them; metabolic depression (diaschisis) and/or degeneration of OR neurons-bearing secondary to central lesions appears therefore as a likely mechanism. Central and peripheral forms of NP may differ in distribution of brain opioid system changes and this in turn might underlie their different sensitivity to opiates. PMID- 17137715 TI - A monoclonal antibody inhibits gelatinase B/MMP-9 by selective binding to part of the catalytic domain and not to the fibronectin or zinc binding domains. AB - Gelatinase B/matrix metalloproteinase-9 (MMP-9) is a multidomain enzyme functioning in acute and chronic inflammatory and neoplastic diseases. It belongs to a family of more than 20 related zinc proteinases. Therefore, the discovery and the definition of the action mechanism of selective MMP inhibitors form the basis for future therapeutics. The monoclonal antibody REGA-3G12 is a most selective inhibitor of human gelatinase B. REGA-3G12 was found to recognize the aminoterminal part and not the carboxyterminal O-glycosylated and hemopexin protein domains. A variant of gelatinase B, lacking the two carboxyterminal domains, was expressed in insect cells and fragmented with purified proteinases. The fragments were probed by one- and two-dimensional Western blot and immunoprecipitation experiments with REGA-3G12 to map the interactions between the antibody and the enzyme. The interaction unit was identified by Edman degradation analysis as the glycosylated segment from Trp(116) to Lys(214) of gelatinase B. The sequence of this segment was analysed by hydrophobicity/hydrophilicity, accessibility and flexibility profiling. Four hydrophilic peptides were chemically synthesized and used in binding and competition assays. The peptide Gly(171)-Leu(187) in molar excess inhibited partially the binding of MMP-9 to REGA-3G12 and thus refines the structure of the conformational binding site. These results define part of the catalytic domain of gelatinase B/MMP-9, and not the zinc-binding or fibronectin domains, as target for the development of selective inhibitors. PMID- 17137716 TI - Mass spectrometric evidence of covalently-bound tetrahydrolipstatin at the catalytic serine of Streptomyces rimosus lipase. AB - We have recently detected that the lipase from Streptomyces rimosus belongs to a large but poorly characterised family of SGNH hydrolases having the alpha beta alpha-fold. Our biochemical characterisation relates to the specific inhibition of an extracellular lipase from Streptomyces rimosus (SRL, 24.2 kDa, Q93MW7) by the preincubation method with tetrahydrolipstatin (THL). In high molar excess (THL/SRL=590 at 25 degrees C, pH=7.0) and after 2 h of incubation in an aqueous system, 56% of the enzyme inhibition was reached. Under the same conditions and in the presence of 50% (v/v) 2-propanol/water, 71% enzyme inhibition was obtained. Kinetic measurements are in agreement with pseudo-first-order kinetics. The nucleophilic attack of the catalytic serine residue 10 of SRL occurs via an opening of the beta-lactone ring of tetrahydrolipstatin and formation of a covalent ester bond. The intact covalent complex of SRL-inhibitor was analysed by ESI and vacuum MALDI mass spectrometry and, furthermore, the exact covalent THL linkage was determined by vacuum MALDI high-energy collision-induced dissociation tandem mass spectrometry. PMID- 17137717 TI - Circulating nucleic acids (CNAs) and cancer--a survey. AB - It has been known for decades that it is possible to detect small amounts of extracellular nucleic acids in plasma and serum of healthy and diseased human beings. The unequivocal proof that part of these circulating nucleic acids (CNAs) is of tumor origin, initiated a surge of studies which confirmed and extended the original observations. In the past few years many experiments showed that tumor associated alterations can be detected at the DNA and RNA level. At the DNA level the detection of point mutations, microsatellite alterations, chromosomal alterations, i.e. inversion and deletion, and hypermethylation of promoter sequences were demonstrated. At the RNA level the overexpression of tumor associated genes was shown. These observations laid the foundation for the development of assays for an early detection of cancer as well as for other clinical means. PMID- 17137718 TI - Sepsis and burn complicated by sepsis alter cardiac transporter expression. AB - Sepsis alone and burn complicated by sepsis produce significant cardiac dysfunction. Since calcium handling by the cardiomyocyte is essential for cardiac function, one mechanism for cardiac abnormalities may be calcium dyshomeostasis. We hypothesized that sepsis and burn plus sepsis alter cardiac calcium transporter expression. Sprague-Dawley rats were divided into: (1) control, (2) sepsis (intratracheal S. Pneumoniae, 4x10(6) CFU), and (3) burn (40% TBSA) plus sepsis. Myocyte [Ca(2+)](i) and [Na(+)](i) were quantified with Fura-2 AM and SBFI, respectively. Western blot analysis of rat hearts used antibodies against the sarcoplasmic reticular Ca(2+) ATPase (SERCA), the L-type calcium channel, the Na(+)/Ca(2+) exchanger or the Na(+)/K(+) ATPase. RESULTS: Sepsis in the presence and absence of burn trauma increased [Ca(2+)](i) and [Na(+)](i). SERCA expression was decreased in the sepsis and burn plus sepsis groups while calcium channel expression was transiently increased in these sepsis groups. Na(+)/Ca(2+) exchanger expression exhibited a biphasic pattern of altered expression. Sepsis and burn plus sepsis reduced Na(+)/K(+) ATPase protein levels. These data suggest that altered transporter expression produce cardiomyocyte calcium and sodium loading and may contribute to sepsis-mediated cardiac contractile dysfunction. PMID- 17137719 TI - Effect of silver on burn wound infection control and healing: review of the literature. AB - Silver compounds have been exploited for their medicinal properties for centuries. At present, silver is reemerging as a viable treatment option for infections encountered in burns, open wounds, and chronic ulcers. The gold standard in topical burn treatment is silver sulfadiazine (Ag-SD), a useful antibacterial agent for burn wound treatment. Recent findings, however, indicate that the compound delays the wound-healing process and that silver may have serious cytotoxic activity on various host cells. The present review aims at examining all available evidence about effects, often contradictory, of silver on wound infection control and on wound healing trying to determine the practical therapeutic balance between antimicrobial activity and cellular toxicity. The ultimate goal remains the choice of a product with a superior profile of infection control over host cell cytotoxicity. PMID- 17137720 TI - Modulation of respiratory activity by locomotion in lampreys. AB - In vertebrates, locomotion is associated with changes in respiratory activity, but the neural mechanisms by which this occurs remain unknown. We began examining this in lampreys using a semi-intact preparation of young adult Petromyzon marinus, in which respiratory and locomotor behaviors can be recorded simultaneously with the activity of the underlying neural control systems. Spontaneous fictive respiration was recorded with suction electrodes positioned over the glossopharyngeal or the rostral vagal motor nucleus. In this preparation, locomotor activity, characterized by symmetrical tail movements (electromyogram recordings), was evoked by mechanical stimulation of the skin. During locomotion, the mean respiratory frequency and the mean area of the motor bursts were significantly increased (81.6+/-28.6% and 62.8+/-25.4%, respectively; P<0.05). The frequency returned to normal 92+/-51 s after the end of locomotion. There were fluctuations in the instantaneous respiratory and locomotor frequencies that were rhythmical but antiphasic for the two rhythmic activities. The changes in respiratory activity were also examined during bouts of locomotion occurring spontaneously, and it was found that a modification in respiratory activity preceded the onset of spontaneous locomotion by 3.5+/-2.6 s. This suggests that the early respiratory changes are anticipatory and are not caused by feedback generated by locomotion. The increase in respiratory frequency during locomotion induced by sensory stimulation persisted after removal of the mesencephalon. When both the mesencephalon and spinal cord were removed, resulting in the isolation of the rhombencephalon, changes in the respiratory activity were also present following skin stimulations that would have normally induced locomotion. Altogether, the results suggest that respiratory changes are programmed to adjust ventilation prior to motor activity, and that a central rhombencephalic mechanism is involved. PMID- 17137721 TI - Thiamine deficiency induces endoplasmic reticulum stress in neurons. AB - Thiamine (vitamin B1) deficiency (TD) causes region selective neuronal loss in the brain; it has been used to model neurodegeneration that accompanies mild impairment of oxidative metabolism. The mechanisms for TD-induced neurodegeneration remain incompletely elucidated. Inhibition of protein glycosylation, perturbation of calcium homeostasis and reduction of disulfide bonds provoke the accumulation of unfolded proteins in the endoplasmic reticulum (ER), and cause ER stress. Recently, ER stress has been implicated in a number of neurodegenerative models. We demonstrated here that TD up-regulated several markers of ER stress, such as glucose-regulated protein (GRP) 78, growth arrest and DNA-damage inducible protein or C/EBP-homologus protein (GADD153/Chop), phosphorylation of eIF2alpha and cleavage of caspase-12 in the cerebellum and the thalamus of mice. Furthermore, ultrastructural analysis by electron microscopic study revealed an abnormality in ER structure. To establish an in vitro model of TD in neurons, we treated cultured cerebellar granule neurons (CGNs) with amprolium, a potent inhibitor of thiamine transport. Exposure to amprolium caused apoptosis and the generation of reactive oxygen species in CGNs. Similar to the observation in vivo, TD up-regulated markers for ER stress. Treatment of a selective inhibitor of caspase-12 significantly alleviated amprolium-induced death of CGNs. Thus, ER stress may play a role in TD-induced brain damage. PMID- 17137722 TI - Amyloid-beta vaccination, but not nitro-nonsteroidal anti-inflammatory drug treatment, increases vascular amyloid and microhemorrhage while both reduce parenchymal amyloid. AB - Vaccination with Abeta(1-42) and treatment with NCX-2216, a novel nitric oxide releasing flurbiprofen derivative, have each been shown separately to reduce amyloid deposition in transgenic mice and have been suggested as potential therapies for Alzheimer's disease. In the current study we treated doubly transgenic amyloid precursor protein and presenilin-1 (APP+PS1) mice with Abeta(1 42) vaccination, NCX-2216 or both drugs simultaneously for 9 months. We found that all treatments reduced amyloid deposition, both compact and diffuse, to the same extent while only vaccinated animals, with or without nonsteroidal anti inflammatory drug (NSAID) treatment, showed increased microglial activation associated with the remaining amyloid deposits. We also found that active Abeta vaccination resulted in significantly increased cerebral amyloid angiopathy and associated microhemorrhages, while NCX-2216 did not, in spite of similar reductions in parenchymal amyloid. Co-administration of NCX-2216 did not attenuate this effect of the vaccine. This is the first report showing that active immunization can result in increased vascular amyloid and microhemorrhage, as has been observed with passive immunization. Co-administration of an NSAID agent with Abeta vaccination does not substantially modify the effects of Abeta immunotherapy. The difference between these treatments with respect to vascular amyloid development may reflect the clearance-promoting actions of the vaccine as opposed to the production-modifying effects proposed for flurbiprofen. PMID- 17137723 TI - Genome dynamics and transcriptional deregulation in aging. AB - Genome instability has been implicated as a major cause of both cancer and aging. Using a lacZ-plasmid transgenic mouse model we have shown that mutations accumulate with age in a tissue-specific manner. Genome rearrangements, including translocations and large deletions, are a major component of the mutation spectrum in some tissues at old age such as heart. Such large mutations were also induced by hydrogen peroxide (H2O2) in lacZ-plasmid mouse embryonic fibroblasts (MEFs) and demonstrated to be replication-independent. This was in contrast to ultraviolet light-induced point mutations, which were much more abundant in proliferating than in quiescent MEFs. To test if large rearrangements could adversely affect patterns of gene expression we PCR-amplified global mRNA content of single MEFs treated with H2O2. Such treatment resulted in a significant increase in cell-to-cell variation in gene expression, which was found to parallel the induction and persistence of genome rearrangement mutations at the lacZ reporter locus. Increased transcriptional noise was also found among single cardiomyocytes from old mice as compared with similar cells from young mice. While these results do not directly indicate a cause and effect relationship between genome rearrangement mutations and transcriptional deregulation, they do underscore the stochastic nature of genotoxic effects on cells and tissues and could provide a mechanism for age-related cellular degeneration in postmitotic tissue, such as heart or brain. PMID- 17137724 TI - Wheel running and fluoxetine antidepressant treatment have differential effects in the hippocampus and the spinal cord. AB - Exercise and antidepressants used independently have been shown to increase hippocampal brain-derived neurotrophic factor (BDNF) and neurogenesis. Despite the fact that patients with depression are often prescribed both, the effects of the exercise and fluoxetine antidepressant treatment used in combination are unknown. Using C57Bl/10 female mice, BDNF protein, insulin-like growth factor 1 (IGF-1) protein and neurogenesis were measured in the hippocampus after 21 days of wheel running, 21 days of fluoxetine antidepressant therapy (daily i.p. injections of 5 mg/kg, 10 mg/kg or 25 mg/kg) and the combination of the two. BDNF protein and cytogenesis/neurogenesis increased in the hippocampus with fluoxetine (high dose), but not wheel running. Hippocampal IGF-1 protein did not change with either treatment. There were no synergistic effects of combining exercise and fluoxetine treatment. Recent reports have also shown that exercise induces molecular mechanisms that benefit the spinal cord and can improve recovery after spinal cord injury (SCI); therefore, we repeated the assays in the spinal cord. Results showed that BDNF, IGF-1 and neurogenesis behave independently in the hippocampus and spinal cord. BDNF protein did not change in the spinal cord with either wheel running or fluoxetine treatment. Spinal cord IGF-1 protein did not change with wheel running, but it decreased with fluoxetine (high dose). Furthermore, spinal cord cytogenesis decreased with fluoxetine treatment. The combined wheel running and fluoxetine groups did not show synergistic results. Thus, the hippocampus and the spinal cord respond in distinct ways to wheel running and fluoxetine, and a prior induction of BDNF, IGF-1 or cytogenesis is unlikely to be the mechanism for wheel running providing a margin of protection against SCI. PMID- 17137725 TI - Pharmacological and molecular characterization of ATP-sensitive K(+) conductances in CART and NPY/AgRP expressing neurons of the hypothalamic arcuate nucleus. AB - The role of hypothalamic ATP-sensitive potassium channels in the maintenance of energy homeostasis has been extensively explored. However, how these channels are incorporated into the neuronal networks of the arcuate nucleus remains unclear. Whole-cell patch-clamp recordings from rat arcuate nucleus neurons in hypothalamic slice preparations revealed widespread expression of functional ATP sensitive potassium channels within the nucleus. ATP-sensitive potassium channels were expressed in orexigenic neuropeptide Y/agouti-related protein (NPY/AgRP) and ghrelin-sensitive neurons and in anorexigenic cocaine-and-amphetamine regulated transcript (CART) neurons. In 70% of the arcuate nucleus neurons recorded, exposure to glucose-free bathing medium induced inhibition of electrical excitability, the response being characterized by membrane hyperpolarization, a reduction in neuronal input resistance and a reversal potential consistent with opening of potassium channels. These effects were reversible upon re-introduction of glucose to the bathing medium or upon exposure to the ATP-sensitive potassium channel blockers tolbutamide or glibenclamide. The potassium channel opener diazoxide, but not pinacidil, also induced a tolbutamide and glibenclamide sensitive inhibition of electrical excitability. Single-cell reverse transcription-polymerase chain reaction revealed expression of mRNA for sulfonylurea receptor 1 but not sulfonylurea receptor 2 subunits of ATP-sensitive potassium channels. Thus, rat arcuate nucleus neurons, including those involved in functionally antagonistic orexigenic and anorexigenic pathways express functional ATP-sensitive potassium channels which include sulfonylurea receptor 1 subunits. These data indicate a crucial role for these ion channels in central sensing of metabolic and energy status. However, further studies are needed to clarify the differential roles of these channels, the organization of signaling pathways that regulate them and how they operate in functionally opposing cell types. PMID- 17137726 TI - Neuron number decreases in the rat ventral, but not dorsal, medial prefrontal cortex between adolescence and adulthood. AB - Neuroimaging studies have established that there are losses in the volume of gray matter in certain cortical regions between adolescence and adulthood, with changes in the prefrontal cortex being particularly dramatic. Previous work from our laboratory has demonstrated that cell death can occur as late as the fourth postnatal week in the rat cerebral cortex. The present study examined the possibility that neuronal loss may occur between adolescence and adulthood in the rat prefrontal cortex. Rats of both sexes were examined during adolescence (at day 35) and young adulthood (at day 90). The volume, neuronal number, and glial number of the medial prefrontal cortex (mPFC) were quantified using unbiased stereological techniques. Neurons were lost from the ventral, but not dorsal, mPFC between adolescence and adulthood, suggesting a late wave of apoptosis that was region-specific. This was accompanied by a decrease in the volume of the female ventral mPFC. In contrast to neuron number, the number of glial cells was stable in the ventral mPFC and increased between adolescence and adulthood in the dorsal mPFC. Sex-specific developmental changes in neuron number, glial number, and volume resulted in sex differences in adults that were not seen during adolescence. The loss of neurons at this time may make the peri-adolescent prefrontal cortex particularly susceptible to the influence of environmental factors. PMID- 17137728 TI - Smoking expectancies in smokers and never smokers: an examination of the smoking Consequences Questionnaire-Spanish. AB - The factor structure of smoking expectancies was examined in daily smokers and never smokers. Participants completed the Smoking Consequences Questionnaire Spanish (SCQ-Spanish; [Cepeda-Benito, A., & Reig-Ferrer, A. (2000). Smoking consequences questionnaire-Spanish. Psychology of Addictive Behaviors, 14, 219 230.]). Using confirmatory factor analysis (CFA), the eight-factor structure of the SCQ-Spanish was replicated in smokers only. Except for beliefs about negative health outcome expectancies, daily smoking rate was strongly associated with all types of smoking outcome expectancies. In comparison to men, women smokers reported greater weight control and negative-affect reduction consequences from smoking. An exploratory factor analysis (EFA) suggested a four-factor structure for the scores of never smokers. The findings represent not only the first cross validation of the SCQ-Spanish but they were also congruent with the notion that first-hand smoking experience is not necessary for the development of multifaceted smoking expectancies. Nonetheless, the finding of a more complex factors structure in smokers than in never smokers is in line with the hypothesis that drug-use expectancies become more complex with drug-use experience. PMID- 17137729 TI - Potential usefulness of sevelamer hydrochloride to improve cardiac outcome in patients with coronary disease and normal renal function. PMID- 17137730 TI - Ehrlich tumor as model to study artificial hyperthyroidism influence on breast cancer. AB - This study used Ehrlich solid tumor as an experimental model for breast cancer to investigate the effects of thyroid hormones and castration on tumor development in adult female mice. Artificial hyperthyroidism was induced in animals, and after a 30-day-treatment, they received subcutaneous injection of neoplastic cells between left plantar cushions. We measured the growth of tumor inoculated in the paws for 10 days at necropsy. Hyperthyroidism induction led to significantly increased tumor size in non-castrated animals, and alterations were less intense in association with artificial hyperthyroidism and castration (p<0.05). Histomorphologic and histomorphometric analyses and neoplastic cell characterization were carried out by measuring nuclear diameter, by evaluating AgNORs, by mitotic count, and by measuring cell proliferation using immunohistochemical marker CDC47. At the end of the experiment, we noted metabolism and a decrease in cell proliferation in groups having received l thyroxine, which were more evident in the non-castrated group (p<0.001). PMID- 17137731 TI - FIP1L1-PDGFRA positive chronic eosinophilic leukemia in Tunisian patients. AB - Hypereosinophilic syndromes (HES) are a heterogenous group of rare disorders characterized by sustained and otherwise unexplained overproduction of eosinophils with organ involvement and consecutive dysfunction. Detection of the FIP1L1-PDGFRA fusion gene or the corresponding cryptic 4q12 deletion in HES supports the diagnosis of chronic eosinophilic leukemia (CEL) and provides a molecular explanation for the pathogenesis of this disorder. We screened seven Tunisian patients fulfilling the WHO criteria of HES for the presence of the FIP1L1-PDGFRA fusion gene using nested reverse transcription polymerase chain reaction on peripheral blood samples. Four of the seven patients were positive for this fusion gene. Sequence analysis revealed a substantial heterogeneity of the fusion transcripts due to the involvement of several FIP1L1 exons. All patients were male. The median age at diagnosis was 24 years (range, 18-50); one patient had a history of hypereosinophilia of more than 10 years. Two patients had clinically important and symptomatic eosinophilic endomyocardial disease with thrombotic events. Splenomegaly was constant in FIP1L1-PDGFRA positive CEL but not in the other HES patients (only 1/3). PMID- 17137732 TI - Illicit drug use in young adults and subsequent decline in general health: the Coronary Artery Risk Development in Young Adults (CARDIA) Study. AB - BACKGROUND: The long-term health consequences of drug use among healthy young adults in the general population are not well described. We assessed whether drug use predicted decline in general self-rated health (GSRH) in a community-based cohort, healthy at baseline. METHODS: A prospective cohort of 3124 young adults (20-32 years old) from four US cities, the Coronary Artery Risk Development in Young Adults Study, was followed from 1987/1988 to 2000/2001. All reported "Good" or better GSRH at baseline, with reassessment in 2000/2001. Drug use in 1987/1988 was as follows: 812 participants were Never Users; 1554 Past Users Only; 503 Current Marijuana Users Only; 255 Current Hard Drug Users (e.g. cocaine, amphetamines, opiates). Analyses measured the association of drug use (1987/1988) with decline to "Fair" or "Poor" GSRH in 2000/2001, adjusting for biological and psychosocial covariates. RESULTS: Reporting health decline were: 7.2% of Never Users; 6.5%, Past Use Only; 7.0%, Current Marijuana Only; 12.6%, Current Hard Drugs (p<0.01). After multivariable adjustment, Current Hard Drug Use in 1987/1988 remained associated with health decline (Odds Ratio (OR), referent Never Use: 1.83, 95% confidence interval (CI) 1.07-3.12). The health decline associated with Current Hard Drugs appeared to be partly mediated by tobacco smoking in 2000/2001, which independently predicted health decline (OR 1.66, 95% CI 1.08-2.50) and weakened the apparent effect of Current Hard Drugs (OR 1.21, 95% CI 0.62-2.36). CONCLUSIONS: Hard drug use in healthy young adults, even when hard drug use stops, is associated with a subsequent decrease in general self rated health that may be partially explained by persistent tobacco use. PMID- 17137733 TI - Improved repeat identification and masking in Dipterans. AB - Repetitive sequences are a major constituent of many eukaryote genomes and play roles in gene regulation, chromosome inheritance, nuclear architecture, and genome stability. The identification of repetitive elements has traditionally relied on in-depth, manual curation and computational determination of close relatives based on DNA identity. However, the rapid divergence of repetitive sequence has made identification of repeats by DNA identity difficult even in closely related species. Hence, the presence of unidentified repeats in genome sequences affects the quality of gene annotations and annotation-dependent analyses (e.g. microarray analyses). We have developed an enhanced repeat identification pipeline using two approaches. First, the de novo repeat finding program PILER-DF was used to identify interspersed repetitive elements in several recently finished Dipteran genomes. Repeats were classified, when possible, according to their similarity to known elements described in Repbase and GenBank, and also screened against annotated genes as one means of eliminating false positives. Second, we used a new program called RepeatRunner, which integrates results from both RepeatMasker nucleotide searches and protein searches using BLASTX. Using RepeatRunner with PILER-DF predictions, we masked repeats in thirteen Dipteran genomes and conclude that combining PILER-DF and RepeatRunner greatly enhances repeat identification in both well-characterized and un annotated genomes. PMID- 17137734 TI - Cyclodextrins and their pharmaceutical applications. AB - Cyclodextrins were first described by Villiers in 1891. Schardinger laid the foundation of the cyclodextrin chemistry in 1903-1911 and identified both alpha- and beta-cyclodextrin. In the 1930s, Freudenberg identified gamma-cyclodextrin and suggested that larger cyclodextrins could exist. Freudenberg and co-workers showed that cyclodextrins were cyclic oligosaccharides formed by glucose units and somewhat later Cramer and co-workers described their ability to form inclusion complexes. By the early 1950s the basic physicochemical characteristics of cyclodextrins had been discovered, including their ability to solubilize and stabilize drugs. The first cyclodextrin-related patent was issued in 1953 to Freudenberg, Cramer and Plieninger. However, pure cyclodextrins that were suitable for pharmaceutical applications did not come available until about 25 years later and at the same time the first cyclodextrin-containing pharmaceutical product was marketed in Japan. Later cyclodextrin-containing products appeared on the European market and in 1997 also in the US. New cyclodextrin-based technologies are constantly being developed and, thus, 100 years after their discovery cyclodextrins are still regarded as novel excipients of unexplored potential. PMID- 17137735 TI - Comparison of cytochrome P450 inhibition assays for drug discovery using human liver microsomes with LC-MS, rhCYP450 isozymes with fluorescence, and double cocktail with LC-MS. AB - The disparity of IC(50)s from CYP450 inhibition assays used to assess drug-drug interaction potential was investigated, in order to have evidence for selecting a reliable in vitro CYP450 inhibition assay to support drug discovery. Three assays were studied: individual rhCYP isozymes and corresponding coumarin derivative probe substrates with fluorescent detection, human liver microsomes (HLM) and cocktail drug-probe substrates with LC-MS detection, and double cocktail rhCYP isozymes mix and drug-probe mix with LC-MS detection. Data comparisons showed that the rhCYP-fluorescent assay and the cocktail assay with HLM-LC-MS had weak correlation. Detection method and probe substrates were shown to not be the major cause of the disparity in IC(50)s. However, the enzyme source and composition (HLM versus, rhCYP) caused disparity in IC(50)s. Specifically, the high concentrations of CYP isozymes often used with HLM-based assays produced high probe substrate conversion and test compound metabolism, which should both contribute to artificially higher IC(50)s. Non-specific binding of substrate to higher concentration proteins and lipids in the HLM-based assays should also contribute to higher IC(50)s. The modified double cocktail assay was found to overcome limitations of the other two assays. It uses an rhCYP isozymes mix, drug probe substrate mix, low protein concentration, and LC-MS detection. The double cocktail assay is sensitive, selective, and high throughout for use in drug discovery to provide an early alert to potential toxicity with regard to drug drug interaction, prioritize chemical series, and guide structural modification to circumvent CYP450 inhibition. PMID- 17137736 TI - Olivocochlear reflex effect on human distortion product otoacoustic emissions is largest at frequencies with distinct fine structure dips. AB - Activity of the medial olivocochlear efferents can be inferred by measuring the change of the level of distortion product otoacoustic emissions (DPOAE) during ipsilateral or contralateral acoustic stimulation, the so-called medial olivocochlear reflex (MOCR). A limitation of this measurement strategy, however, is the distinct variability of MOCR values depending on DPOAE primary tone levels and frequency, which makes selection of the stimulus parameters difficult. The objective of this study was to evaluate the dependence of MOCR values on DPOAE fine structure in humans. MOCR during contralateral acoustic stimulation was measured at frequencies with distinct non-monotonicity ("dip") in the DPOAE fine structure, and in frequencies with flat fine structure. One hundred and twenty one different primary tone level combinations were used (L(1)=50-60dB SPL, L(2)=35-45dB SPL, 1dB steps). The measurement was repeated on another day. The major findings were: (1) Largest MOCR effects can be found in frequencies which exhibit a distinct dip in DPOAE fine structure. (2) Primary tone levels have a critical influence on the magnitude of the MOCR effect. MOCR changes of up to 23dB following a L(1) change of only 1dB were observed. Averages of the maximum MOCR change per 1dB step were in the 3-5dB-range. Both findings can be interpreted in the light of the DPOAE two-generator model [Heitmann, J., Waldmann, B., Schnitzler, H.U., Plinkert, P.K., Zenner, H.P. 1998. Suppression of distortion product otoacoustic emissions (DPOAE) near 2f1-f2 removes DP-gram fine structure - evidence for a secondary generator. Journal of the Acoustical Society of America 103, 1527-1531]. According to the present results we propose, that assessing MOCR specifically at frequencies with a distinct dip in the DPOAE fine structure, in combination with fine variation of the stimulus tone levels, allows for a more targeted search for maximum MOCR effects. Future studies must show if this approach can contribute to the further clarification of the physiological roles of the olivocochlear efferents. PMID- 17137737 TI - An ethnobotanical survey of medicinal plants used in Loja and Zamora-Chinchipe, Ecuador. AB - This paper reports the results of an ethnobotanical survey on the uses of medicinal plants by inhabitants of two southern Ecuadorian provinces, namely, Loja and Zamora-Chinchipe. In this region, two surviving ethnic groups, the Saraguros and the Shuars, and the descendants of a now extinct culture, the Paltas, have been identified. The present study reports a total of 275 plant species, having 68 different therapeutical uses. PMID- 17137738 TI - Voice rehabilitation with Provox2 voice prosthesis following total laryngectomy for laryngeal and hypopharyngeal carcinoma. AB - OBJECTIVE: To analyze the effectiveness of the Provox2 voice prosthesis for voice rehabilitation following total laryngectomy. METHODS: From September 2000 to December 2004, the Provox2 voice prosthesis was used for voice rehabilitation in 32 patients following total laryngectomy. The quality of speech with the Provox2 voice prosthesis was analyzed using the HRS rating scale, the maximum phonation time (MPT), incidence of complications and the in situ lifetime. The rate of speech restoration was further analyzed in 129 patients with total laryngectomy from 1996 to 2004. RESULT: Twenty-nine of 32 patients were able to restore speech using the Provox2 voice prosthesis, a speech restoration rate of 90.6%. The maximum phonation time (MPT) was measured in 18 patients using the Provox2 voice prosthesis. The mean MPT was 15.1 s, with a range of 8-28 s. MPT was not influenced by age, concurrent radiotherapy treatment, the location of the primary tumor or use of reconstructive surgery. The average lifetime of the Provox2 in patients with laryngeal carcinoma (12 patients) and hypopharyngeal carcinoma (17 patients) was 27.2 and 16.6 weeks, respectively, which was significantly different (P=0.024, non-parametric Mann-Whitney's U-test). The rate of speech restoration by the use of esophageal speech, and insertion of an artificial larynx was 62.7% for laryngeal carcinoma (59 cases) and 38.6% for hypopharyngeal carcinoma (70 cases), which was also significantly different (P<0.01, chi-square test). CONCLUSION: Provox2 voice prosthesis speech was very useful due to the higher rate of speech restoration, longer phonatory time, and better intelligibility. It was also thought that voice prosthesis speech was useful in conjunction with esophageal speech and an artificial larynx depending on the patient's condition or wishes. PMID- 17137739 TI - [Follow-up criteria for community acquired pneumonias and acute exacerbations of chronic obstructive pulmonary disease]. AB - The follow-up of Community Acquired Pneumonias (CAP) and Acute Exacerbations of Chronic Obstructive Pulmonary Diseases (AECOPD) differs with the setting of care, but overall calls upon the same investigations as the initial evaluations. In the event of initial ambulatory care, the evaluation is carried out primarily on clinical data, at the 2 or 3rd day for the CAP, at the 2nd to 5th day for the AECOPD. In the event of unfavourable evolution, or from the start in the most severe cases, the follow-up is carried out in hospital; clinical evaluation is readily daily, and all the more frequent that the clinical condition is worrying because of the severity or risk factors. The investigations will be limited to those initially abnormal in the event of favourable evolution; on the contrary, unfavourable evolution can justify new investigations which depend on clinical characteristics. Remotely, i.e. 4 to 8 weeks later, must be checked the return at the baseline clinical state, a chest X-ray (CAP), spirometry and arterial blood gas (AECOPD), even bronchoscopy and thoracic CT-scan. PMID- 17137740 TI - [Management of lower respiratory tract infections in immunocompetent adults (community-acquired pneumonia and acute exacerbation of chronic obstructive pulmonary disease): The challenge of a new consensus conference]. PMID- 17137741 TI - Coevolution of exceptional longevity, exceptionally high metabolic rates, and mitochondrial DNA-coded proteins in mammals. AB - Mammals' longevity is inversely related to mass-specific basal metabolic rate because the generation of reactive oxygen species constrains lifespan. Longevity increases with body mass because the latter is inversely related to mass-specific basal metabolic rates. In placental mammals the longevity residuals from the power laws that describe longevity as a function of mass-specific basal metabolic rates, or body mass, are positively correlated with the relative rates of evolution of cytochrome b, a generator of reactive oxygen species. Therefore, longevity is more accurately described as a function of both mass-specific basal metabolic rate and the relative rate of cytochrome b evolution. The longevity residuals from the power law that describe longevity as a function of body mass are positively correlated with the relative rate of evolution of most other mtDNA coded proteins. In taxa with very high rate of cytochrome b evolution exceptional longevity is associated with an increase, rather than the predicted decrease, of basal metabolic rates. These finding are compatible with the hypothesis that, in placental mammals, the accelerated evolution of mtDNA-coded proteins, allowed the extension of lifespan by selecting mutations that reduce the generation of reactive oxygen species, mostly by increasing internal proton leak, that accelerates mitochondrial electron transport. PMID- 17137742 TI - Parent-adolescent communication about sexuality: the role of adolescents' beliefs, subjective norm and perceived behavioral control. AB - OBJECTIVE: To examine to what extent adolescents' beliefs, subjective norm and perceived behavioral control predict the amount of parent-adolescent communication about sexuality. In addition, the role of adolescents' gender, ethnic background, religiosity and educational level on these relationships was assessed as well. METHODS: Data were collected from 481 students of four high schools in The Netherlands. A questionnaire assessed adolescents' beliefs, subjective norm, perceived behavioral control, and self-reported parent adolescent communication about sexuality. Linear regression analyses were performed to identify predictors of parent-adolescent communication. To assess differences between subgroups, chi(2)-analyses, t-tests and analyses of variance were conducted. RESULTS: Being female and having positive beliefs about talking with parents about sexuality were positively related to amount of parent adolescent communication. In addition, adolescents' perceived behavioral control and subjective norm were significant predictors as well. CONCLUSION: Adolescents' beliefs, subjective norm and perceived behavioral control, are all significant predictors of frequency of parent-adolescent sex communication, with beliefs being the most important. In addition, adolescents' gender predicted a significant amount as well. PRACTICE IMPLICATIONS: Interventions aimed to increase the amount of parent-adolescent communication should primarily target their efforts to changing adolescents' underlying beliefs about discussing sexuality with their parents. Our results furthermore suggest that it is important to take into consideration gender variations in these beliefs, by designing separate interventions for different groups of adolescents. PMID- 17137743 TI - Mortality versus survival graphs: improving temporal consistency in perceptions of treatment effectiveness. AB - OBJECTIVE: Previous research has demonstrated that people perceive treatments as less effective when survival graphs show fewer years of data versus more data. We tested whether using mortality graphs would reduce this temporal inconsistency bias. METHODS: A demographically diverse sample of 1461 Internet users read about a hypothetical disease and then were randomized to view either survival or mortality graphs that showed either 5 years of data or 15 years of treatment outcomes data. Participants identified the most effective treatment, provided ratings comparing the effectiveness of two treatments, and answered comprehension questions. RESULTS: Treatment effectiveness ratings varied significantly between respondents seeing the 5 year and 15 year survival graphs even though the relative risk reduction was the same in both cases. This variation was significantly reduced in the mortality graph conditions. Responses on comprehension measures were mixed: viewers of mortality graphs were less able to identify which treatment was more effective but better able to correctly report individual data points. CONCLUSIONS: Perceptions of treatment effectiveness appear more temporally consistent with mortality graphs than with survival graphs. PRACTICE IMPLICATIONS: All line-based risk graphics (whether framed in survival or mortality terms) should highlight duration information to facilitate improved comprehension of treatment effectiveness. PMID- 17137744 TI - Vitality, mental health, and satisfaction with information after breast cancer. AB - OBJECTIVE: The purpose of this study was to investigate the relationship between satisfaction with information about treatment-related and survivorship issues and mental health outcomes, including vitality, in long-term breast cancer survivors. METHODS: Participants who had finished treatment for breast cancer at least 3 months before enrollment completed a survey instrument designed to evaluate satisfaction with diagnostic and treatment information and satisfaction with survivorship information. Mental health and vitality were measured using the Medical Outcomes Study SF-36, and distress was measured using the impact of events scale. Bivariate analyses and linear regression analyses were performed to investigate the relationships between satisfaction with information, mental health, vitality, and distress controlling for clinical and treatment variables. RESULTS: The survey was completed by 231 participants (response rate 83%). More respondents were highly satisfied with treatment information (87%) than with survivorship information (30%, p=0.0001). There was a strong positive relationship between satisfaction with information (both treatment and survivorship issues) and vitality, mental health, and a strong negative relationship with distress. In multivariate analyses, satisfaction with treatment information was independently associated with mental health (p<0.01), and satisfaction with survivorship information was independently associated with vitality (p<0.05). CONCLUSION: Among patients who have completed treatment for breast cancer, satisfaction with diagnosis and treatment information is greater than satisfaction with survivorship issues and satisfaction with information may play an important role in mental health outcomes. PRACTICE IMPLICATIONS: Given the relationship between satisfaction with information and vitality, interventions to improve informational support regarding survivorship issues are warranted. PMID- 17137745 TI - Interrelationships between negative energy balance (NEB) and IGF regulation in liver of lactating dairy cows. AB - In dairy cows, negative energy balance (NEB) during the early post-partum period is associated with major alterations in the growth hormone-insulin-like growth factor (GH-IGF) axis. Since the liver mediates nutrient partitioning during lactation, we aimed to determine how NEB alters the endocrine regulation of the insulin-like growth factor (IGF) system by investigating the expression of IGF family members and related steroid receptors. On the second day of lactation, cows were allocated to one of two treatments designed to produce mild (MNEB) or severe NEB (SNEB). MNEB cows (n=5) were fed ad lib grass silage supplemented with concentrate and milked x1 daily and SNEB cows (n=6) were restricted in dietary intake and milked x3 daily. Energy balance (EB) status was monitored until the second week of lactation when plasma and liver samples revealed a markedly divergent metabolic profile. At this time, plasma protein and hepatic mRNA for IGF-I was reduced in SNEB cows compared with MNEB cows. Both levels of expression correlated highly when data from all animals was pooled (r=0.963; P<0.01). SNEB cows also exhibited reduced hepatic expression for transcripts encoding IGF-1R, IGF-2R, IGF binding proteins (IGFBPs) -3, -4, -5, -6, acid labile subunit, and receptors for oestrogen (ERalpha) and growth hormone (total GHR and 1A variant), while the expression of IGFBP-2 was elevated. Expression of mRNA for IGF-II, IGFBP-1 and receptors for insulin (A/B) and glucocorticoid (alpha) was unaffected by EB. Results demonstrate that SNEB affects hepatic synthesis of IGF-I, and other components known to modulate the bioavailability and stability of circulating IGF-I. PMID- 17137746 TI - Prediction in breast cancer of the extent of axillary node involvement from the size and lymphovascular invasion status of the primary tumour: medico-legal considerations. AB - AIM: To ascertain for medico-legal purposes in the United Kingdom, the extent to which in breast cancer primary tumour size and lympho-vascular invasion (LVI) status determine axillary node staging. METHODS: Four hundred and fifty symptomatic patients had their tumour sizes stratified into 1cm ranges and the percentage incidence/probability of the axillary stage being 1, 2 or 3 was calculated separately for those tumours that showed no evidence of LVI and those that did. RESULTS: The presence of LVI has a marked effect on the likelihood and extent of axillary node involvement for all primary tumour sizes. CONCLUSIONS: This needs to be recognized and quantified when prediction of this extent is made for medico-legal purposes. Within the constraints of the 'balance of probabilities' argument in UK law, it is rarely possible to differentiate between the likelihood of the axillary stage being 2 or 3. PMID- 17137747 TI - Struggles for equivalence: in vitro developmental toxicity model evolution in pharmaceuticals in 2006. AB - Our group has been using the ECVAM Embryonic Stem Cell assay to predict developmental toxicity. In order to improve the separation of non-teratogens from weak teratogens, we have employed measures of gene expression, and different statistical methods from those originally used to develop the test. These approaches have fundamentally not improved the discrimination of 'weaks' from 'nons'. A realization that a very low value for cytotoxicity IC50 would drive a final result for the test in ways that were inappropriate for pharmaceuticals has led us to re-examine the cytotoxicity component. Our current efforts are focused on other, perhaps more sensitive, measures of cytotoxicity, combined with gene expression changes in mouse stem cells in an attempt to correctly identify weak teratogens and non-teratogens. PMID- 17137748 TI - Lambda-cyhalothrin-induced changes in oxidative stress biomarkers in rabbit erythrocytes and alleviation effect of some antioxidants. AB - Erythrocytes are a convenient model to understand the membrane oxidative damage induced by various xenobiotic-prooxidants. This study was designed to investigate (1) the possibility of lambda-cyhalothrin (LC), a type II pyrethroid, to induce oxidative stress response in rabbit erythrocytes in vitro and its effect on selected antioxidant enzymes and (2) the role of vitamin C (VC; 20mM) and vitamin E (VE; 2mM) in alleviating the cytotoxic effects of LC. Erythrocytes were divided into three groups. The first group, previously prepared erythrocytes was incubated for 4h at 37 degrees C with different concentrations (0, 0.1, 0.5, 1, 2.5, 5mM) of LC. The second and third groups were preincubated with VC or VE, respectively for 20 min and followed by LC incubation for 4h. Following in vitro exposure, LC caused a significant induction of oxidative damage in erythrocytes at different concentrations as evidenced by increased thiobarbituric acid reactive substances (TBARS) levels. However, a significant decrease in the content of sulfhydryl groups (SH-groups), and the activities of acetylcholinesterase (AChE), superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) were observed. The response was concentration dependent. VC or VE pretreated erythrocytes showed a significant protection against the cytotoxic effects induced by LC on the studied parameters. In conclusion, antioxidant vitamins especially VE could be able to ameliorate LC induced oxidative stress by decreasing lipid peroxidation and altering antioxidant defense system in erythrocytes. PMID- 17137749 TI - Multiple co-localizations in arcuate GHRH-eGFP neurons in the mouse hypothalamus. AB - In the present work, we took advantage of a recently described model of GHRH enhanced green fluorescent protein (eGFP) transgenic mice to evaluate the extent of co-localization of GHRH neurons with galanin (GAL), neurotensin (NT) and tyrosine hydroxylase (TH) in 3- and 8-month-old male and female mice. The total number of GHRH-eGFP neurons along the rostro-caudal axis of the arcuate nucleus did not differ according to gender or age. GAL-immunoreactivity was present in 40 44% of 3-month-old GHRH-eGFP neurons in male and female arcuate nucleus, respectively, but only 25-22% in 8-month-old mice. TH immunoreactivity occurred in 36-35% of GHRH-eGFP neurons in male and female arcuate nucleus from 3-month old mice and these proportions increased to 40 and 45% in 8-month-old mice. NT immunoreactivity was present in 14 and 24% of GHRH-eGFP neurons in male and female arcuate nucleus from 3-month-old mice up to 28 and 26% in 8-month-old mice. Thus, co-localization of peptides and enzyme in GHRH-eGFP neurons displays a sexual dimorphism at 3-month of age for NT, and at 8-month for TH, while the total number of GHRH-eGFP neurons does not exhibit gender difference at either age. In summary, it appears that changes in co-localized (and presumably co released) peptides, rather than GHRH per se, may contribute to the changes in sexually dimorphic GH secretion with aging in the mouse. PMID- 17137750 TI - Effects of amphetamine on serotoninergic and GABAergic expression of developing brain. AB - Roles of age and withdrawal were explored in mechanisms underlying the action of amphetamine (Amph), by monitoring the serotonergic and GABAergic expression in key brain regions of the rat. Postnatal 21 and 60 day-old male rats were intraperitoneally injected with D-Amph, 5 mg/kg, or saline, three times daily for 14 days and then withdrawn from Amph for 0 or 14 days; these animals received single injections on day 15 (W0d) or day 29 (W14d). Following Amph injections, though both age groups exhibited hyperlocomotion, stereotypy and behavioral sensitization, the juvenile showed 100-300% longer latencies to reach and 30%-42% shorter duration of maximal behavioral scores than the adult from day 2-29. Immunocytochemical analysis revealed down-regulation of 42-76% in 5 hydroxytryptamine (HT) immunoreactive processes in motor and somatosensory cortices, and hippocampus of both ages after Amph exposure at W0d. At W14d, the 5 HT resembled saline-control in the Amph-treated juvenile, whereas remained weakened in the adult. By contrast, densities of GAD67 (glutamic acid decarboxylase)-boutons were up-regulated by 35-545% in the neocortical areas, nucleus accumbens, caudate-putamen and hippocampus of all Amph-administered rats. After 14 days withdrawal, the juvenile recovered the decreased 5-HT fibers, but not the increased GABAergic, indicating unique roles of the two systems in response to Amph. PMID- 17137751 TI - The role of nitric oxide on the convulsive behavior and oxidative stress induced by methylmalonate: an electroencephalographic and neurochemical study. AB - Methylmalonic acidemias consist of a group of inherited metabolic disorders caused by deficiency of methylmalonyl-CoA mutase activity and biochemically characterized by methylmalonate (MMA) accumulation, impairment mitochondrial oxidative metabolism and reactive species production. Preliminary studies with nitric oxide synthase (NOS) inhibitors have suggested that nitric oxide (NO) plays a role in the convulsant effect of MMA. However, definitive biochemical and electrophysiological evidence of the involvement of NO in the convulsions induced by MMA are lacking. In this study, we investigated whether the inhibition of NOS by 7-nitroindazole (7-NI, 3-60mg/kg, i.p.) altered the convulsions, protein oxidative damage, NO(x) (NO(2) plus NO(3)) production and Na(+),K(+)-ATPase activity inhibition induced by MMA. 7-NI decreased striatal NO(x) content, but increased seizures and protein carbonylation induced by MMA (6mumol/striatum). The intrastriatal injection of l-arginine (50nmol/0.5mul), but not of d-arginine (50nmol/0.5mul), increased striatal NO(x) content and protected against MMA induced electroencephalographic seizures, striatal protein carbonylation and Na(+),K(+)-ATPase inhibition. Furthermore, l-arginine (50nmol/0.5mul) and MMA had no additive effect on NO(x) increase. These results are experimental evidence that endogenous NO plays a protective role in the convulsions and acute neurochemical alterations induced by this organic acid. PMID- 17137752 TI - The activity of protease inhibitors against Giardia duodenalis and metronidazole resistant Trichomonas vaginalis. AB - Antiretroviral protease inhibitors were assessed in vitro for their activity against Giardia duodenalis and Trichomonas vaginalis. Kaletra (a co-formulation of ritonavir and lopinavir) was the most effective overall, with 50% effective drug concentrations (EC(50)) of 1.1-2.7 microM (ritonavir concentration) against G. duodenalis and 6.8-8 microM against metronidazole-sensitive and clinically metronidazole-resistant T. vaginalis. Minimal inhibitory concentrations were 2 2.5 microM and 10-50 microM for G. duodenalis and T. vaginalis, respectively. Within the range of human plasma concentrations for ritonavir, only G. duodenalis was inhibited. Lopinavir alone was less inhibitory than ritonavir but was associated with a blockage in cytokinesis of G. duodenalis trophozoites. Saquinavir was not effective. These findings are significant considering the association between human immunodeficiency virus and T. vaginalis, and between G. duodenalis and homosexual behaviour. PMID- 17137753 TI - The 50th anniversary of the discovery of 6-aminopenicillanic acid (6-APA). AB - This year (2007) marks the 50th anniversary of [corrected] discovery of 6 aminopenicillanic acid (6-APA), the precursor of all semi-synthetic penicillins [corrected] This review, by a scientist who played a major part in the discovery and a physician who participated in the early clinical trials of these antibiotics, tells the story of the discovery and of the early development of the beta-lactam antibiotics that revolutionised the treatment of infections. PMID- 17137754 TI - Structure-antimicrobial activity relationship for silanols, a new class of disinfectants, compared with alcohols and phenols. AB - Triorganosilanols (R(CH(3))(2)SiOH) were recently reported to exhibit unexpectedly strong disinfectant properties. The antimicrobial activities of silanols were significantly higher than their analogous alcohols. A study of the structural dependence of their antimicrobial activity was conducted against four bacteria, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Enterococcus faecalis. Silanols, alcohols with structures analogous to the silanols (R(CH(3))(2)COH) and substituted phenols were evaluated as a single class of materials. The minimum lethal concentrations (MLCs), defined as the concentration required for a 7-log reduction in viable bacteria after 1h exposure, were used to measure antimicrobial activity. Octanol-water partition coefficients (logP) and hydrogen bond acidities (deltanu), measured as the shift in frequency of the OH stretching band between free OH and hydrogen-bonded OH to diethyl ether oxygen by infrared spectroscopy, were utilised as dispersive and polar structural parameters, respectively. The correlation established by multiple regression analysis between antimicrobial activities and structural properties of silanols, alcohols and phenols against the four bacteria treated as a single family produced the following equation, log(1/MLC)=0.679 logP+0.0036deltanu-1.909 (n=282, r=0.96, s=0.22). This equation and the significantly high correlation coefficient supported the hypothesis that the lipophilic properties and the H-bond acidities are primary factors for the antimicrobial action of silanols, alcohols and phenols. The high antimicrobial activity of silanols is explained by their greater H-bond acidity and their enhanced lipophilicity. PMID- 17137755 TI - Intercountry transfer of PER-1 extended-spectrum beta-lactamase-producing Acinetobacter baumannii from Romania. PMID- 17137756 TI - Implementation and evaluation of a once-daily amikacin dosing protocol in a long term care facility. PMID- 17137757 TI - Gender differences in facial emotion recognition in persons with chronic schizophrenia. AB - BACKGROUND: The aim of the present study was to investigate possible sex differences in the recognition of facial expressions of emotion and to investigate the pattern of classification errors in schizophrenic males and females. Such an approach provides an opportunity to inspect the degree to which males and females differ in perceiving and interpreting the different emotions displayed to them and to analyze which emotions are most susceptible to recognition errors. METHODS: Fifty six chronically hospitalized schizophrenic patients (38 men and 18 women) completed the Penn Emotion Recognition Test (ER40), a computerized emotion discrimination test presenting 40 color photographs of evoked happy, sad, anger, fear expressions and neutral expressions balanced for poser gender and ethnicity. RESULTS: We found a significant sex difference in the patterns of error rates in the Penn Emotion Recognition Test. Neutral faces were more commonly mistaken as angry in schizophrenic men, whereas schizophrenic women misinterpreted neutral faces more frequently as sad. Moreover, female faces were better recognized overall, but fear was better recognized in same gender photographs, whereas anger was better recognized in different gender photographs. CONCLUSIONS: The findings of the present study lend support to the notion that sex differences in aggressive behavior could be related to a cognitive style characterized by hostile attributions to neutral faces in schizophrenic men. PMID- 17137758 TI - Effects of frequent cannabis use on hippocampal activity during an associative memory task. AB - Interest is growing in the neurotoxic potential of cannabis on human brain function. We studied non-acute effects of frequent cannabis use on hippocampus dependent associative memory, investigated with functional Magnetic Resonance Imaging (fMRI) in 20 frequent cannabis users and 20 non-users matched for age, gender and IQ. Structural changes in the (para)hippocampal region were measured using voxel-based morphometry (VBM). Cannabis users displayed lower activation than non-users in brain regions involved in associative learning, particularly in the (para)hippocampal regions and the right dorsolateral prefrontal cortex, despite normal performance. VBM-analysis of the (para)hippocampal regions revealed no differences in brain tissue composition between cannabis users and non-users. No relation was found between (para)hippocampal tissue composition and the magnitude of brain activity in the (para)hippocampal area. Therefore, lower brain activation may not signify neurocognitive impairment, but could be the expression of a non-cognitive variable related to frequent cannabis use, for example changes in cerebral perfusion or differences in vigilance. PMID- 17137759 TI - Three-year antipsychotic effectiveness in the outpatient care of schizophrenia: observational versus randomized studies results. AB - Antipsychotic discontinuation rates are a powerful indicator of medication effectiveness in schizophrenia. We examined antipsychotic discontinuation in the Schizophrenia Outpatient Health Outcomes (SOHO) study, a 3-year prospective, observational study in outpatients with schizophrenia in 10 European countries. Patients (n=7728) who started antipsychotic monotherapy were analyzed. Medication discontinuation for any cause ranged from 34% and 36% for clozapine and olanzapine, respectively, to 66% for quetiapine. Compared to olanzapine, the risk of treatment discontinuation before 36 months was significantly higher for quetiapine, risperidone, amisulpride, and typical antipsychotics (oral and depot), but similar for clozapine. Longer medication maintenance was associated with being socially active and having a longer time since first treatment contact for schizophrenia, whereas higher symptom severity, treatment with mood stabilizers, substance abuse, having hostile behaviour were associated with lower medication maintenance. Antipsychotic maintenance in SOHO was higher than the results of previous randomized studies. PMID- 17137760 TI - Photoinduced phospholipid polymer grafting on Parylene film: advanced lubrication and antibiofouling properties. AB - Poly(p-xylylene) (Parylene C) coatings have been applied in implantable electronic devices because of their electrical insulation and moisture barrier properties. To provide lubrication and an antibiofouling surface, a biomimetic phospholipid polymer--poly(2-methacryloyloxyethyl phosphorylcholine (MPC))--was grafted from the surface using UV irradiation with benzophenone as an initiator. The poly(MPC) grafting on the Parylene C films was confirmed by attenuated total reflection-Fourier transfer inflated irradiation, X-ray photoelectron spectroscopy and ellipsometry. These analyses indicated that the Parylene C films were completely covered by the poly(MPC)-graft layer with an average thickness of 140 nm under dry condition. The atomic force microscope (AFM) images revealed that the poly(MPC)-graft chains extended under wet condition. However, they formed globular structures under dry condition. Water contact angle measurements revealed a decreased receding angle of 29.5 degrees on the poly(MPC)-grafted surface with a high hysteresis of 41.4 degrees. These results indicate that the poly(MPC)-graft chains gain mobility in a wet environment. The average kinetic friction coefficient of the poly(MPC)-grafted surface in water was 0.018, which was 90% lower than that of the original surface. The in vitro single protein adsorption reduced by over 70% due to the poly(MPC) grafting. The hydrated poly(MPC)-graft chains are considered to provide lubrication and antibiofouling properties. The surface zeta potential measurement clarified the electroneutrality of the poly(MPC)-grafted surface. We concluded that the poly(MPC) grafting from the Parylene C layer significantly improved its surface properties and, subsequently, its biological properties. PMID- 17137761 TI - Study of the cholesterol-GM3 ganglioside interaction by surface pressure measurements and fluorescence microscopy. AB - The nature of the cholesterol/glycolipid interaction in rafts being poorly understood, the interaction of cholesterol with the GM(3) ganglioside has been studied by surface pressure measurements and fluorescence microscopy. Results have been compared to those obtained with sphingomyelin (SM)-cholesterol and palmitoyl-oleoyl-phosphatidylcholine (POPC)-cholesterol monolayers. The analysis of (pi-A) isotherms of mixed monolayers show a condensing effect of cholesterol on GM(3) molecules, in the same range than the effect observed with POPC and higher than the effect on SM. This is likely due to the similar state of GM(3) and POPC, since both molecules are in liquid expanded phases in our experimental conditions. The study of the cholesterol desorption induced by beta-cyclodextrin suggests also that the GM(3)-cholesterol interaction is rather weak as in the case of POPC-cholesterol interaction, and clearly lower than SM-cholesterol one. This lack of interaction is discussed in terms of nature of lipid chains and molecular shape, and suggests that no hydrogen bond is formed between GM(3) and cholesterol polar heads. Fluorescence microscopy performed on mixed GM(3) cholesterol monolayers shows the presence, at surface pressure higher than 10 mN/m, of particular blurring patterns without defined boundary, which could be due to a partial solubilization in one phase of different phases observed at lower surface pressure, whereas SM-cholesterol and POPC-cholesterol monolayers are homogeneous at the lateral resolution of our microscopy set-up. PMID- 17137762 TI - Surface sliding friction of negatively charged polyelectrolyte gels. AB - The friction between two polyelectrolyte gels carrying the same or opposite sign of charges has been investigated using a rheometer. It is found that the friction was strongly dependent on the interfacial interaction between two gel surfaces. In the repulsive interaction case, especially, the friction was extremely low. The friction behavior is attempted to be described in terms of the hydrodynamic lubrication of the solvent layer between two like-charged gel surfaces, which is formed due to the electrostatic repulsion of the two gel surfaces. From the theoretical analysis (hydrodynamic mechanism), the friction behaviors were explained qualitatively, all of the experimental results, nevertheless, could not be understood well. The viscoelastic feature of the gel and the non-Newtonian behavior of water at the friction interface are considered to be important to elucidate the gel friction. PMID- 17137763 TI - Structure modification in hen egg yolk low density lipoproteins layers between 30 and 45 mN/m observed by AFM. AB - We have studied the structure of films made by low density lipoproteins (LDL) from hen egg yolk, which are composed of apoproteins, neutral lipids and phospholipids. These LDL have been deposited on air-water interface to form a monolayer which has been compressed to measure an isotherm using Langmuir balance. This isotherm presented three transitions (neutral lipid (surface pressure, pi=19 mN/m), apoprotein-lipid (pi=41 mN/m) and phospholipid (pi=51 mN/m) transitions). We have studied only the apoprotein-lipid transition. In order to observe the LDL film structure before (pi=30 mN/m) and after (pi=45 mN/m) the apoprotein-lipid transition, the formed films were transferred and visualised by atomic force microscopy (AFM). Our results have shown that the structures observed in the LDL film were different depending on the surface pressure. The apoproteins and neutral lipids appeared to be miscible up to the apoprotein-lipid transition, when demixing occurred. The structures observed after the apoprotein-lipid transition should be due to the demixing between apoproteins and neutral lipids. On the other hand, apoproteins and phospholipids seemed miscible whatever the surface pressure. Hence, the first transition (pi=19 mN/m) should be attributed to the free neutral lipid collapse; the second transition (pi=41 mN/m) should be attributed to the demixing of apoprotein neutral lipid complexes; and the last transition (pi=51 mN/m) should be attributed to phospholipid collapse or to demixing of apoprotein-phospholipid complexes. PMID- 17137764 TI - Incorporation of water-soluble drugs in PLGA microspheres. AB - Poly(lactide-co-glycolide) (PLGA) microspheres containing blue dextran, as a model of water-soluble drugs, were prepared from w(1)/o/w(2) emulsions by using a microhomogenizer and a solvent evaporation method. Effects of preparation conditions, such as, concentration of poly(vinyl alcohol) (PVA) in w(2) phase, viscosity of inner soluble water phase, volume ratio of oil phase to w(1) phase in primary emulsion, PLGA concentration in oil phase, and molecular weight or composition of PLGA, upon the properties of PLGA microspheres containing water soluble drugs were examined. Concentration of poly(vinyl alcohol) (PVA), the dispersant dissolved in w(2) phase of secondary emulsion did not show any effects on the final particle size. On the other hand, volume ratio of oil phase to water one in primary emulsion affected the final particle size, which seemed to be related to the local PLGA concentration in w(1)/o emulsions. That is, the particle size increased as the volume ratio of w(1) phase against oil phase, w(1)/o (v/v), increased. The loading efficiency, however, was not affected by the volume ratio of w(1)/o (v/v), but affected by blue dextran concentration in w(1) phase. Higher loading efficiency was observed in PLGA microspheres prepared from w(1) phase containing lower concentration of blue dextran. Blue dextran solution (inner water phase) with the lower viscosity may result in the lower leakage ratio of blue dextran during the preparation procedure. Increases in concentration and molecular weight of PLGA made particle size larger. PMID- 17137765 TI - [Malignant solid tumors in neonates: a study of 71 cases]. AB - Malignant neonatal tumors are rare and comprise 2% of childhood malignancies. Clinical features, histologic types, prognosis were very different from those seen in older children, facing oncologists with diagnostic, therapeutic and ethical problems. PATIENTS AND METHODS: In a retrospective study from January 1987 to January 2004, we reviewed the management of neonates treated at the Institute Gustave Roussy for a malignant solid tumor for whom symptoms started in the first month of life. RESULTS: Seventy-one neonates were treated, comprising 1,2% of the overall patients treated during the same period of time. Of these 71 patients, 42 (59%) presented with neuroblastomas, 12 (17%) with mesenchymal tumors, 6(8%) with cerebral tumors and 11 with various other types of tumors. Fifty-nine patients underwent surgical resection. Thirty-eight neonates received chemotherapy, administered at a 30 to 50% reduced dose. Hematologic toxicities and infections were the main therapeutic complications. Very small doses of radiotherapy were used in only 5 children. There has been no therapy-related mortality. Twenty-two of the 57 survivors have sequelae, especially patients with intraspinal neuroblastoma. The 5 year overall survival was 79%. CONCLUSIONS: Neonatal malignant solid tumors, except for cerebral tumors, have a good prognosis. The young age of patients resulted in problems of treatment tolerance. The therapeutic regimen should take into account the risk of acute iatrogenic toxicity and long term sequelae. Surgery remains the treatment of choice but chemotherapy, with dose reduction, managed by expert teams, is essential and safer in a lot of case. PMID- 17137766 TI - [Prevalence of Pneumocystis jiroveci pneumonia in Tunisian primary immunodeficient patients]. AB - Pneumocystis Jiroveci pneumonia (PJP) is a rare opportunistic infection in immunodeficient patients in Tunisia, as well as in other Africain countries including those with a high prevalence of AIDS. In the literature, PJP has been reported in primary immunodeficiency diseases (PID) namely SCID T-B- or T-B+ or X linked hyper-IgM syndrome. OBJECTIVE: To evaluate the prevalence of PJP in the different PID observed in Tunisia. PATIENTS AND METHODS: This retrospective study concerned 290 cases of PID confirmed by immunological investigation including the study of specific and/or non-pecific humoral and cellular immunity. The identification of P. Jiroveci in patients suspected of pneumocystosis was achieved by parasitological investigation in bronchoalveolar lavages. RESULTS: A PID associated to a parasitologically confirmed pneumocystic infection was found in 9 out of 290 patients (3%) among whom the majority (7 patients) had an HLA class II combined immunodeficiency. The latter is an autosomic recessive disease which has been reported mainly in North African families. Indeed, this population is characterized by a high rate of consanguinity. Interestingly, no PJP has been observed neither in SCID T-B- or T-B+ nor in X-linked hyper-IgM syndrome. DISCUSSION AND CONCLUSION: PJP seems to be particularly frequent in HLA class II deficiency patients, since 7 out of 22 patients with this deficiency had a PJP (31%). Due to this defect, antigen presenting cells are unable to present the antigen to T lymphocytes demonstrating the critical role of CD4+ T lymphocytes responses in the immune response to this pathogen. PMID- 17137767 TI - [Etidronate per os in subcutaneous fat necrosis with hypercalcemia and nephrocalcinosis]. AB - Subcutaneus fat necrosis is an uncommon disease which may be complicated with potentially fatal hypercalcemia. The usual treatment of hypercalcemia includes hyperhydratation, corticosteroids and diet. This treatment is not always effective in normalizing plasma calcium concentration. A treatment with bisphosphonates has been effective in similar cases. We report on the case of a patient presenting with symptomatic hypercalcemia, complicated with nephrocalcinosis consecutive to subcutaneus fat necrosis after birth asphyxia at term. Oral etidronate has been used for 3 weeks, after failure of classical treatment. The evolution was favorable. After a short review of the complications of subcutaneus fat necrosis, we discuss the use of bisphophonates in this indication. PMID- 17137768 TI - [Thrombotic thrombocytopenic purpura in a newborn]. AB - A newborn presented with haemolytic anemia, thrombocytopenia, hyperbilirubinemia and renal failure as early as the first hours of life. An early plasmatherapy was undertaken, followed by good outcome. The specific von Willebrand factor-cleaving protease (ADAMTS 13) was found at less than 5%. This is the specific biologic diagnostic element of congenital thrombotic thrombocytopenic purpura or Upshaw Schulman syndrome. This disease of constitutional thrombotic microangiopathy was well identified and understood only few years ago. It's a rare disease which early diagnosis and treatment are crucial in order to preserve functional and vital capacities of the patient. PMID- 17137769 TI - [Non epileptic paroxysmal movement disorders in childhood]. AB - Paroxysmal movement disorders are not uncommon in childhood, but are probably under-recognised. Paroxysmal movement disorders are a distinctive group of disorders that represents various clinical situations, characterised by intermittent and episodic disturbances of movement. Diagnosis relies on semiological analysis, mainly based on parental description of the manifestations; video recording (during an EEG-video monitoring or home made video) are often helpful to establish the correct diagnosis. In the large majority of the cases, paroxysmal movement disorders are benign situations. Some of them are transient, as they spontaneously stop over time (benign torticolis of infancy, paroxysmal tonic upgaze). Being familiar with these disorders will lead to accurate diagnosis, so avoiding useless investigations. Most of the time, no treatment will be required, and the families will be informed of the good prognosis. PMID- 17137770 TI - [Self-produced locomotion and spatial cognition: a new light from spinal muscular atrophy]. AB - Various studies have shown that occurrence of locomotion in infancy is correlated with the development of visuospatial cognitive competencies, suggesting that locomotor experience might play a central role in spatial development, especially in the realm of manual search for hidden objects. However, recent studies indicate that young children with spinal muscular atrophy (SMA), a hereditary neuromuscular disease which results in severe motor impairments, excel in some spatial cognitive skills. Indeed, striking cognitive performances are exhibited by young SMA children in some areas such as the ability to search successfully for hidden objects and the acquisition of the spatial vocabulary. The performances of SMA children suggest that, despite their total deprivation of locomotor experience, they have the capacity to acquire and use rich spatial representations. As a result, locomotor impairment does not appear to be a key risk factor for dramatic slowing down or deviation in the development of spatial search skills. PMID- 17137771 TI - Insulin glargine improves glycemic control and health-related quality of life in type 1 diabetes. AB - BACKGROUND AND AIMS: Glargine improves glucose control and reduces the risk of nocturnal hypoglycemia compared with neutral protamine Hagedorn (NPH) insulin. To date, only one study has measured the effects of glargine on health-related quality of life (HRQL); the aim of this study was therefore to confirm the beneficial effects of glargine on disease-specific HRQL in type 1 diabetes. METHODS AND RESULTS: Forty-seven patients (mean age, 46 range, 25-74; males, 54%) with diabetes of at least 1-year duration, and with suboptimal glucose control under intensive insulin treatment (IIT), were switched from NPH to glargine. Forty patients maintained on IIT were used as controls. Diabetes-related HRQL was assessed using the Well-being Enquiry for Diabetics (WED), before and after a 6- to 8-month switch to glargine. An 11-item questionnaire based upon diabetes specific issues was used to assess treatment satisfaction and perceived changes after switching. On glargine, the mean glycosylated hemoglobin decreased by 0.7% (treatment vs. baseline, P<0.0001) and several WED scores improved (discomfort, P=0.020; impact, P=0.0002; total score, P=0.0005). WED changes were associated with a lower perceived risk of hypoglycemia and less problems in daily life on glargine. CONCLUSIONS: The results of this study show that the beneficial effect of glargine is not limited to better metabolic control; the burden of type 1 diabetes mellitus on everyday life is also reduced. PMID- 17137773 TI - Extraction and composition of three naturally occurring anti-cancer alkaloids in Camptotheca acuminata seed and leaf extracts. AB - Naturally occurring camptothecins (CPT) are important sources of chemotherapeutic agents for clinical treatment of cancer. Extraction of CPT from Camptotheca acuminata trees remains to be a cost-effective way in the supply equation compared with a total synthesis. This study conducted a series of experiments to determine efficient solvent for the maximal extraction of CPT and its two derivatives, hydroxycamptothecin (HCPT) and methoxycamptothecin, from seeds and leaves of C. acuminata. Methanol as an extraction solvent demonstrated in seeds a significantly higher recovery of these three alkaloids than dichloromethane and acetone. Methanol concentrations at 70% in water resulted in maximum extraction of all the three alkaloids regardless of the type of plant materials. However, other strengths of methanol, lower or higher, either decreased the extracting power or showed no improvement in the extraction. Seed extract contained all the three alkaloids whereas leaf extract was absent of HCPT. A stable ratio of the three alkaloids was discovered but it was dependent upon seed or leaf extract of C. acuminata, which with various compositions can be produced. Ecological and medicinal implications of the leaf and seed extract characterized with different chemical compositions are discussed. PMID- 17137775 TI - Transforming growth factor-beta: innately bipolar. AB - Widely heralded for depressing ongoing immune responses, renewed interest in the proficiency by which transforming growth factor beta (TGF-beta) not only engages but also might drive an over-reactive innate response highlights its bipolar nature. Although coordination of the development and function of Treg, in addition to direct inhibition of cellular activation, are prominent pathways by which TGF-beta controls adaptive immunity, paradoxically TGF-beta appears instrumental in initiation of host responses to invasion through recruitment and activation of immune cells and persuasion of Th17 lineage commitment. Nevertheless, true to its manic-depressive behavior, new evidence links TGF-beta with depression of innate cells, including NK cells, and by way of a potential bridge between mast cells and Treg. Disruption of the tenuous balance between these opposing actions of TGF-beta underlies immunopathogenicity. PMID- 17137774 TI - Laminin-5 activates extracellular matrix production and osteogenic gene focusing in human mesenchymal stem cells. AB - We recently reported that laminin-5, expressed by human mesenchymal stem cells (hMSC), stimulates osteogenic gene expression in these cells in the absence of any other osteogenic stimulus. Here we employ two-dimensional liquid chromatography and tandem mass spectrometry, along with the Database for Annotation, Visualization and Integrated Discovery (DAVID), to obtain a more comprehensive profile of the protein (and hence gene) expression changes occurring during laminin-5-induced osteogenesis of hMSC. Specifically, we compare the protein expression profiles of undifferentiated hMSC, hMSC cultured on laminin-5 (Ln-5 hMSC), and fully differentiated human osteoblasts (hOST) with profiles from hMSC treated with well-established osteogenic stimuli (collagen I, vitronectin, or dexamethazone). We find a marked reduction in the number of proteins (e.g., those involved with calcium signaling and cellular metabolism) expressed in Ln-5 hMSC compared to hMSC, consistent with our previous finding that hOST express far fewer proteins than do their hMSC progenitors, a pattern we call "osteogenic gene focusing." This focused set, which resembles an intermediate stage between hMSC and mature hOST, mirrors the expression profiles of hMSC exposed to established osteogenic stimuli and includes osteogenic extracellular matrix proteins (collagen, vitronectin) and their integrin receptors, calcium signaling proteins, and enzymes involved in lipid metabolism. These results provide direct evidence that laminin-5 alone stimulates global changes in gene/protein expression in hMSC that lead to commitment of these cells to the osteogenic phenotype, and that this commitment correlates with extracellular matrix production. PMID- 17137776 TI - Species of colletotrichum on agavaceae. AB - Species of Colletotrichum cause diseases on a wide range of hosts, frequently infecting plants in the Agavaceae (monocotyledons: Liliales). Three species of Colletotrichum restricted to the Agavaceae were detected through morphological studies of specimens and molecular sequence analyses of the LSU of the nu-rDNA and the ITS region of the nu-rDNA from cultures. Colletotrichum agaves on Agave is fully described and illustrated. Colletotrichum dracaenophilum is described as a new species for isolates having long conidia and occurring on Dracaena sanderiana from China. Colletotrichum phormii and Glomerella phormii are determined to be the correct scientific names for the asexual and sexual states, respectively, of a species commonly referred to as C. rhodocyclum and G. phacidiomorpha occurring mainly on Phormium. In addition, C. gloeosporioides and C. boninense were isolated from plants in the Agavaceae. All species of Colletotrichum described on Agavaceae were evaluated based on type specimens. A key to the five species of Colletotrichum on Agavaceae is included. This paper includes one new species, Colletotrichum dracaenophilum, and three new combinations, Colletotrichum phormii, Glomerella phormii, and Phaeosphaeriopsis phacidiomorpha. PMID- 17137777 TI - Increased myocardial vulnerability and autonomic nervous system imbalance in obstructive sleep apnea syndrome. AB - BACKGROUND: Obstructive sleep apnea syndrome (OSAS) is characterized by the repeated episodes of upper airway obstruction during sleep, leading to significant hypoxia. Noninvasive evaluation of autonomic nervous system (ANS) and myocardial vulnerability may help determination of OSAS patients who are under high risk of malignant cardiac arrhythmias. The aim of this study was to show the effects of OSAS on predictors of arrhythmias by the evaluation of heart rate turbulence (HRT), heart rate variability (HRV) and QT dynamicity reflecting the ANS balance and myocardial vulnerability. METHODS: After polysomnographic study, 80 patients with OSAS and 55 age matched OSAS (-) subjects were included in the study. Twenty-four-hour Holter monitoring was performed in all subjects. HRT, HRV and QT dynamicity parameters were calculated. RESULTS: Turbulence slope was significantly decreased in OSAS patients whereas turbulence onset was increased (P<0.001). QT/RR slopes were significantly increased for QT end and QT apex (P<0.001). In HRV analysis, autonomic balance changed in favor of sympathetic system at night in OSAS patients. Furthermore, HRT and QT dynamicity parameters are found to be correlated with Apnea-Hypopnea Index (AHI). CONCLUSION: OSAS is associated with a significant worsening in HRV, HRT, and QT dynamicity parameters. Our results may indicate that HRV and QT dynamicity parameters can be useful noninvasive methods that may detect autonomic nervous system activity and ventricular vulnerability in OSAS. PMID- 17137778 TI - Expression of chemokine receptors CXCR3 and CCR4 in lymphocytes of idiopathic nonspecific interstitial pneumonia. AB - Little is known about the role of chemokines and their receptors interaction, which are essential for recruitment of selective lymphocyte subsets during inflammation, in the pathogenesis of idiopathic nonspecific interstitial pneumonia (NSIP). Recent studies have revealed Th1 and Th2 cells preferentially employ the chemokine receptors, CXCR3 and CCR4, respectively, in the process of accumulation into inflammatory sites. We evaluated the CXCR3 and CCR4 expression on infiltrated lymphocytes in lung tissues of 12 NSIP cases and 10 idiopathic pulmonary fibrosis (IPF) cases in our previous study. The number of CXCR3 positive lymphocytes of NSIP patients was significantly greater than that of IPF patients (261.1+/-145.1 vs. 64.9+/-27.0, P<0.01). The number of CCR4 positive lymphocytes of NSIP patients was significantly lower than that of IPF (9.5+/-8.3 vs.62.6+/-26.9, P<0.01). The CXCR3 to CCR4 ratio of NSIP patients was significantly greater than that of IPF patients (47.9+/-45.9 vs. 1.11+/-0.40, P<0.01). The differences of CXCR3 positive, CCR4 positive lymphocyte counts, and of CXCR3/CCR4 ratio between cellular and fibrosing NSIP were not significant. These results suggest that a Th1 pattern of chemokine receptor expression predominates in the lung interstitium of patients with NSIP but, in IPF patients, CCR4 might be relatively predominant, in contrast to the finding in NSIP patients, and that Th1/Th2 balance might be an important factor in the pathogenesis of NSIP. PMID- 17137779 TI - Guidelines of the French Speaking Society for Chest Medicine for management of malignant pleural mesothelioma. AB - Previously considered as a rare tumor, malignant pleural mesothelioma (MPM) has become a very important public health issue. In fact, MPM is a tumor with a poor survival, and its incidence is expected to continue to increase for at least the next 10 years. Asbestos exposure is the main factor involved in MPM pathogenesis. The diagnosis of MPM may be difficult because of differential diagnosis such as pleural benign disease induced by asbestos exposure or pleural metastasis of adenocarcinoma. Management of patients with MPM also remains complicated because they are often referred for evaluation late in the evolution of the disease. Moreover, MPM exhibits a high resistance to radiotherapy and chemotherapy; only few patients are candidates for radical surgery. New therapeutic strategies such as gene or cell therapy are still on clinical trial. Therefore, an optimal treatment of MPM is not clearly defined yet, despite the introduction of recent drugs. Between April 2005 and January 2006, the French Speaking Society for Chest Medicine (SPLF), in collaboration with other French scientific societies, brought together experts on mesothelioma to draw up recommendations in order to provide clinicians with clear, concise, up-to-date guidelines on management of MPM, presented in this report. PMID- 17137780 TI - Development of a glucose-6-phosphate biosensor based on coimmobilized p hydroxybenzoate hydroxylase and glucose-6-phosphate dehydrogenase. AB - This work reports the development of an amperometric glucose-6-phosphate biosensor by coimmobilizing p-hydroxybenzoate hydroxylase (HBH) and glucose-6 phosphate dehydrogenase (G6PDH) on a screen-printed electrode. The principle of the determination scheme is as follows: G6PDH catalyzes the specific dehydrogenation of glucose-6-phosphate by consuming NADP(+). The product, NADPH, initiates the irreversible the hydroxylation of p-hydroxybenzoate by HBH in the presence of oxygen to produce 3,4-dihydroxybenzoate, which results in a detectable signal due to its oxidation at the working electrode. The sensor shows a broad linear detection range between 2 microM and 1000 microM with a low detection limit of 1.2 microM. Also, it has a fast measuring time which can achieve 95% of the maximum current response in 20s after the addition of a given concentration of glucose-6-phosphate with a short recovery time (2 min). PMID- 17137781 TI - Larvicidal activities against Aedes aegypti of some Brazilian medicinal plants. AB - Larvicidal activities against Aedes aegypti have been determined in the ethanolic extracts obtained from 51 Brazilian medicinal plants. Eleven of the 84 extracts studied showed significant (LC50 < 100 microg mL(-1)) activities against larvae, with extracts from Annona crassiflora (root bark, LC50 = 0.71 microg mL(-1); root wood, LC50 = 8.94 microg mL(-1)) and Annona glabra (seed, LC50 = 0.06 microg mL( 1)) showing the highest activities. The results obtained should be of value in the search for new natural larvicidal compounds. PMID- 17137782 TI - Enzymatic deinking of laser printed office waste papers: some governing parameters on deinking efficiency. AB - The protocol for the enzymatic deinking of laser printed waste papers on a laboratory scale using cellulase (C) and hemicellulase (H) of Aspergillus niger (Amano) was developed as an effective method for paper recycling. A maximum deinking efficiency of almost 73% by the enzyme combination of C:H was obtained using the deinking conditions of pulping consistency of 1.0% (w/v) with the pulping time of 1.0min, temperature of 50 degrees C, pH=3.5, agitation rate of 60rpm, pulp concentration of 4% (w/v), concentration of each enzyme of 2.5U/g air dried pulp and the enzyme ratio of 1:1. The deinking efficiency was further enhanced to 95% using the optimized flotation system consisting of pH=6.0, Tween 80 of concentration 0.5% (w/w), working air flow rate of 10.0L/min and temperature of 45 degrees C. The deinked papers were found to exhibit properties comparable to the commercial papers suggesting the effectiveness of the enzymatic process developed. PMID- 17137783 TI - The influence of ambient lighting levels on postural sway in healthy children. AB - The purpose of this study was to determine whether postural sway in healthy children varied in different levels of ambient lighting. Twelve boys and 26 girls with a mean age of 118 months stood on a force platform under three conditions: eyes closed, eyes opened in regular light (200 lx) and eyes opened in dim light (3 lx). Analysis of variance and pairwise comparisons revealed significantly more postural sway with the eyes closed condition compared to the regular and dim light conditions but no differences between the regular and dim light conditions. While our results on postural sway during the eyes closed condition are consistent with current findings in the pediatric and adult literature, our findings comparing postural sway during regular and dim light conditions differ from those found in older adults. It appears that the visual system of children is efficient in dim light conditions, adding support to the view that quiet standing is more dependent on vision in older adults than in younger individuals. PMID- 17137784 TI - Bone mineral density of the proximal tibia relates to axial torsion in the lower limb. AB - This study sought to determine the relationship between bone mineral density distribution in the proximal tibia and tibial and femoral torsions, hip-knee ankle angle, hip rotation index (internal-external rotation), knee abduction moments, and the foot progression angle. Simple linear regression found that the hip rotation index (r=-0.59, p<0.001), tibial torsion (r=-0.41, p=0.004), and knee abduction moments (r=0.39, p=0.005) were significantly related to medial lateral BMD ratio for all subjects. The three variables were then studied together to determine their relationship to the bone mineral distribution in the proximal tibia using multiple linear regression (r=0.80, p<0.001). These findings suggest that higher medial knee joint loads, loss of internal rotation of the hip, and internal tibial torsion may lead to increased medial versus lateral BMD of the proximal tibia of healthy knees. Similar bone distribution patterns are found in knees with osteoarthritis; therefore, we suggest these may be risk factors. PMID- 17137785 TI - Nonhost-associated proliferation of intrahyphal hyphae of citrus scab fungus Elsinoe fawcettii: refining the perception of cell-within-a-cell organization. AB - Ultrastructural aspects of intrahyphal hyphae formation were investigated in Elsinoe fawcettii by transmission electron microscopy. Desiccated and hydrated cultures of E. fawcettii hyphae in liquid and solid media were prepared to determine the effects of water/nutrient availability and media fluidity on the formation of intrahyphal hyphae of the fungus. In all the culture conditions, intrahyphal hyphae were observed in enclosing hyphae. Electron-transparent hyphal cell walls clearly delimited intrahyphal hyphae from the cytoplasm of enclosing hyphae. Intrahyphal hyphae occupied most of the lumen of the enclosing hyphae, and showed intact hyphal cytoplasm with distinct organelles and inclusions. Intrahyphal hyphae were found to grow out of the degenerated hyphae that were almost devoid of cellular contents (simple intrahyphal hyphae). Some intrahyphal hyphae appeared to push aside a septum and passed into the adjacent hyphal cell. Besides a single intrahyphal hypha, instances were noted where enclosing hyphae contained several individual intrahyphal hyphae (multiple intrahyphal hyphae). Other enclosing hyphae contained intrahyphal hyphae, which also had intrahyphal hyphae (compound intrahyphal hyphae). The cell wall of intrahyphal hyphae showed the continuity with the cell wall of the enclosing hyphae. Concentric bodies typical of ascomycetes occurring in dry habitats were not found in all the types of hyphae. These results suggest that intrahyphal hyphae formation of E. fawcettii does not require plant defense responses. The fungus is thought to form intrahyphal hyphae during the saprophytic phase in ex planta ecological niches as well as the parasitic phase in host parts. PMID- 17137786 TI - Preparation and biological studies of 125I-DOTA-TATE. AB - DOTA-TATE, a somatostatin analog was radiolabeled with (125)I in good yields and high radiochemical purity. The product exhibited good stability in vitro. Pharmacokinetic studies in normal Swiss mice showed rapid blood clearance with low thyroid uptake. Biodistribution studies in murine melanoma showed 3.0+/-1.3% ID/g uptake in tumor at 3h post injection (p.i.), with negligible reduction at 24h p.i. Inhibition studies carried out in vivo using cold DOTA-TATE confirmed the tumor specificity of the product. PMID- 17137787 TI - Purification of a beta-galactosidase from cotyledons of Hymenaea courbaril L. (Leguminosae). Enzyme properties and biological function. AB - Beta-galactosidases are enzymes that can be found in most living beings and in the plant kingdom its activity and genes have been detected in several tissues such as ripening fruits, developing leaves and flowers and storage tissues such as cotyledons. In plants, their activities are usually associated with the secondary metabolism or with oligosaccharide or polysaccharide degradation. Polysaccharide specific beta-galactosidases include beta-galactanases, which attack pectic polymers and beta-galactosidases that attack xyloglucans (XG). In the present work we purified an XG-specific beta-galactosidase (named hcbetagal) from cotyledons of developing seedlings of Hymenaea courbaril, a legume tree from the Neotropical region of the world. The enzyme has a molecular weight of 52-62 kDa and was shown to attack specifically xyloglucan oligosaccharides (XGOs) but not the polymer. It has a pH optimum between 3 and 4 and at this pH range the enzyme increases activity linearly up to 50 degrees C. Kinetic studies showed that hcbetagal is inhibited competitively by free galactose (K(i) = 3.7). The biochemical properties of hcbetagal as a whole suggest that it is involved in storage xyloglucan mobilisation during seedling development. Its high specificity towards XGOs, the low pH optimum and the fact that it is inhibited by its product (galactose) suggest that hcbetagal might be one of the biochemical control points in xyloglucan catabolism in vivo. A possible relationship with functional stability of the wall during cell death as cotyledons undergo senescence is discussed. PMID- 17137788 TI - Management of congenital tracheal stenosis in infancy. PMID- 17137789 TI - How to clamp the main pulmonary artery during video-assisted thoracoscopic surgery lobectomy. AB - We sometimes run across difficulty in dissection of the pulmonary arteries due to dense pleural adhesions and bleeding from the pulmonary artery during the video assisted thoracoscopic surgery (VATS) lobectomy. In these cases, conversion of the VATS approach to open thoracotomy is a requisite. The presence of an easy and safe technique for pulmonary artery clamping will make the switch of the surgical procedure unnecessary. We developed the new technique for pulmonary artery clamping using 1-0 silk suture. This may become one of the standard techniques for pulmonary artery clamping not only in VATS but also in open thoracotomy, as well. PMID- 17137790 TI - Spatial genome organization in the formation of chromosomal translocations. AB - Chromosomal translocations and genomic instability are universal hallmarks of tumor cells. While the molecular mechanisms leading to the formation of translocations are rapidly being elucidated, a cell biological understanding of how chromosomes undergo translocations in the context of the cell nucleus in vivo is largely lacking. The recent realization that genomes are non-randomly arranged within the nuclear space has profound consequences for mechanisms of chromosome translocations. We review here the emerging principles of spatial genome organization and discuss the implications of non-random spatial genome organization for the genesis and specificity of cancerous chromosomal translocations. PMID- 17137792 TI - Oxidative burst in hard clam (Mercenaria mercenaria) haemocytes. AB - Haemocytes of bivalve molluscs are known to be responsible for many immunological functions, including recognition, phagocytosis, and killing or elimination of invading microorganisms, such as potentially infective bacteria and parasites. In many bivalves, killing of microorganisms engulfed by haemocytes is accomplished by a sudden release of reactive oxygen species (ROS) within the haemocytes; this response is referred to as an oxidative burst. Previous studies have failed to detect oxidative burst in haemocytes of the hard clam (northern quahog), Mercenaria mercenaria. In the present study, we applied a widely used chemical probe for ROS detection in haemocytes, dichlorofluorescin-diacetate (DCFH-DA), to haemocytes from this clam species and used flow cytometry to quantify fluorescence in individual haemocytes. Oxidation of DCFH-DA to the fluorescent product, DCF, within unstimulated haemocytes indicated that ROS were clearly produced in these cells. Two activators of oxidative burst, zymosan and bacterial extracellular products, which have been applied successfully to haemocytes in other species, stimulated large increases in ROS production in hard clam haemocytes. Furthermore, two inhibitors of ROS production, W-13 and diphenylene iodinium (DPI), significantly suppressed ROS production by haemocytes. Nitric oxide synthase inhibitors, NMMA and L-NIO, did not suppress ROS production, indicating that the observed oxidation of DCFH-DA is not mediated by nitric oxide. These results show unequivocally that haemocyte oxidative burst is active in M. mercenaria and, therefore, is a likely mechanism in host response to pathogens and parasites. PMID- 17137791 TI - SR protein 9G8 modulates splicing of tau exon 10 via its proximal downstream intron, a clustering region for frontotemporal dementia mutations. AB - The microtubule-associated protein tau is important to normal neuronal function in the mammalian nervous system. Aggregated tau is the major component of neurofibrillary tangles (NFTs), present in several neurodegenerative diseases, including Alzheimer's and frontotemporal dementia with Parkinsonism (FTDP). Splicing misregulation of adult-specific exon 10 results in expression of abnormal ratios of tau isoforms, leading to FTDP. Positions +3 to +16 of the intron downstream of exon 10 define a clustering region for point mutations that are found in FTDP. The serine/arginine-rich (SR) factor 9G8 strongly inhibits inclusion of tau exon 10. In this study, we established that 9G8 binds directly to this clustering region, requires a wild-type residue at position +14 to inhibit exon inclusion, and RNAi constructs against 9G8 increase exon 10 inclusion. These results indicate that 9G8 plays a key role in regulation of exon 10 splicing and imply a pathogenic role in neurodegenerative diseases. PMID- 17137793 TI - Haemocyte reactions in WSSV immersion infected Penaeus monodon. AB - White spot syndrome virus (WSSV) has been a major cause of shrimp mortality in aquaculture worldwide in the past decades. In this study, WSSV infection (by immersion) and behaviour recruitment of haemocytes is investigated in gills and midgut, using an antiserum against the viral protein VP28 and a monoclonal antibody recognising haemocytes (WSH8) in a double immunohistochemical staining and in addition transmission electron microscopy was applied. More WSH 8(+) haemocytes were detected at 48 and 72 h post-infection in the gills of infected shrimp compared to uninfected animals. Haemocytes in the gills and midgut were not associated with VP28-immunoreactivity. In the gills many other cells showed virus replication in their nuclei, while infected nuclei in the gut cells were rare. Nevertheless, the epithelial cells in the midgut showed a clear uptake of VP28 and accumulation in supranuclear vacuoles (SNV) at 8h post-infection. However, epithelial nuclei were never VP28-immunoreactive and electron microscopy study suggests degradation of viral-like particles in the SNV. In contrast to the gills, the midgut connective tissue shows a clear increase in degranulation of haemocytes, resulting in the appearance of WSH8-immunoreactive thread-like material at 48 and 72 h post-infection. These results indicate recruitment of haemocytes upon immersion infection in the gills and degranulation of haemocytes in less infected organs, like the midgut. PMID- 17137794 TI - Global activation of primary motor cortex during voluntary movements in man. AB - Unilateral voluntary movements are accompanied by robust activation of contralateral primary motor cortex (M1) in a somatotopic fashion. Occasionally, coactivation of M1 (M1-CoA) ipsilateral to the movement was described. In a study with brain tumor patients, we consistently observed additional somatotopic M1 CoAs and hypothesized that they might represent a basic feature of movement execution. To test this hypothesis, we used BOLD functional magnetic resonance imaging in healthy subjects and show that unilateral voluntary movements of the fingers or toes go along not only with contralateral M1 activation, but also with ipsilateral M1-CoA of the respective homotopic representation and bilateral M1 CoA of different heterotopic representations not directly involved in the executed movement. Moreover, bilateral M1-CoA of heterotopic representations was observed in tongue movements. All M1-CoAs respected the correct somatotopy; however, their Euclidean coordinates were shifted and resembled to those obtained for imagined movements rather than for actual movements. BOLD signal intensities and correlations to the applied hemodynamic reference function were lower in M1 CoAs as compared to the M1 activations driving the movement but did not differ between homo- and heterotopic M1-CoAs. Thus, we propose that specific unilateral voluntary movements are accompanied by a global activation of primary motor areas, reflecting an overall increase in neuronal activity and unraveling the fundamental principle of distributed processing in M1. Executive motor function may rely on a balance of inhibitory and excitatory neuronal activity, where actual movement would result from a shift towards excitation. PMID- 17137795 TI - Observation of two distinct spatial-temporal BOLD clusters during sensory stimulation in rats. AB - Neuronal activity evokes changes in local CBF and CBV, whose spatial differences are not fully known. We use the Radial Correlation Contrast (RCC) analysis method with high spatial resolution 100 x 100 x 1000 microm3 data collected with an 11.7 T magnet to differentiate two spatial-temporal BOLD clusters during sensory rat forepaw stimulation and hypothesize that each corresponds to either the CBF or the CBV processes. One cluster, obtained during the time segment of stimulation onset, is characterized by a high positive BOLD signal whereas the other, obtained during the simulation decline time segment, is characterized by a lower positive signal and strong post stimulus undershoot. The average volume of stimulation onset clusters is embedded in the stimulation decline clusters with the latter significantly larger and shifted towards deeper cortical layers. Comparison of amplitude-RCC and cross-correlation analyses performed on equivalent time segments (30 s, 40 images) revealed no differences in cluster size or location, demonstrating that temporal locality is more important than spatial locality in distinguishing between stimulation onset and stimulation decline clusters. We hypothesize that clusters characterized by stimulation onset are highly weighted by local changes in CBF whereas clusters characterized by stimulation decline are more CBV weighted. Moreover, the data suggest that the locations of the highest CBF changes are distinct from the locations of the highest CBV changes. While the former located within stimulation decline clusters and its weight is gradually reduced towards cluster's periphery (mainly ventrally), the highest changes in CBV occur in the cluster's periphery with only modest changes towards its center. PMID- 17137796 TI - Cloned Vero cell lines transfected with full-length A-segment or ORF1 cDNA sequence of IBDV. AB - Recombinant plasmids containing the A-segment or VP2/4/3 gene of infectious bursal disease virus (IBDV) were transfected into Vero cells. Monoclonal Vero cell lines were generated under G418 selection. Genomic (PCR/Southern blot) and transcriptional (RT-PCR/Northern blot) analyses showed that one copy of A-segment or VP2/4/3 or a partial gene of them was randomly and stably inserted into genomic DNA of Vero cells, and was able to transcribe corresponding mRNA. IFA/IPMA and Western blot analyses further confirmed that two of the monoclonal Vero cells with insertion of the A-segment of IBDV into genomic DNA could stably express VP2, VP3 and VP5 proteins, one cell line only expressed VP2 protein, and three monoclonal Vero cell lines with genomic insertion of the VP2/4/3 gene of IBDV could express VP2, VP3 and VP4 proteins. Under G418 selection, integrated foreign genes can be inherited along with cellular genomic DNA during cell replications. Moreover, DNA fragmentation and caspase-3 activity assays illustrated that cell apoptosis did not develop in monoclonal Vero cell lines expressing VP2 and VP5 proteins. The monoclonal IBDV gene-inserted Vero cell lines developed in this study will facilitate better understanding of IBDV and other members of the Birnaviridae in an expression system that would enable investigation of virus-host cell interactions on the cellular and molecular level. PMID- 17137797 TI - Muscle actin isoforms are differentially expressed in human satellite cells isolated from donors of different ages. AB - Myogenesis is mainly sustained by a subpopulation of myogenic cells known as satellite cells (SC). In this paper we studied alpha-smooth muscle (alphaSMA) and alpha-sarcomeric muscle (alphaSRA) actin isoform expression in cultures of human satellite cells (HSC) isolated from skeletal muscle biopsies from a 5-day-old newborn, a 34-year-old young adult and a 71-year-old donor. Myogenicity of cultures was assessed using immunocytochemical detection of desmin and myosin heavy chain. Time-course expression of alphaSRA and alphaSMA were studied with both immunocytochemistry and western blotting procedures. Although alphaSMA was never detected in whole skeletal muscle, both alphaSMA and alphaSRA were detected in proliferating and differentiating HSC derived from donors of all examined ages. The expression level experiments showed that alphaSRA was gradually up regulated during HSC differentiation, but no significant differences were observed between newborn, young, and elderly HSC cultures. Our data demonstrated that HSC, isolated from subjects of different ages, re-expressed alphaSMA, but its levels and expression pattern varied considerably in the newborn with respect to the young adult and elderly donors. These results are discussed in relation to the myogenic differentiation capability of HSC during human muscle senescence. PMID- 17137799 TI - Expression of interleukin-10 in intestinal lymphocytes detected by an interleukin 10 reporter knockin tiger mouse. AB - To identify interleukin-10 (IL-10)-producing cells in vivo, we generated a knockin mouse where an internal ribosome entry site (IRES) green fluorescence protein (GFP) element was inserted immediately before the polyadenylation site of the IL-10 gene. GFP fluorescence in cells from these mice was found to correlate positively with IL-10 protein expression. With this model, we found that after multiple T cell receptor (TCR) stimulations, strong expression of IL-10 was produced specifically by intraepithelial lymphocytes (IEL) in the small intestine and colonic lamina propria lymphocytes (cLPL). We found that anti-CD3 treatment induces T regulatory cell 1 (Tr1)-like cells in small intestinal IEL (sIEL) and led to the accumulation of naturally occurring regulatory T (nTreg) cells in colonic LPL (cLPL). These findings highlight the intestine as a unique site for induction of IL-10-producing T cells, which play a critical role in the regulation of inflammation in the gut. PMID- 17137798 TI - Ndfip1 protein promotes the function of itch ubiquitin ligase to prevent T cell activation and T helper 2 cell-mediated inflammation. AB - Nedd4 family interacting protein-1 (Ndfip1) is a protein whose only known function is that it binds Nedd4, a HECT-type E3 ubiquitin ligase. Here we show that mice lacking Ndfip1 developed severe inflammation of the skin and lung and died prematurely. This condition was due to a defect in Ndfip1(-/-) T cells. Ndfip1(-/-) T cells were activated, and they proliferated and adopted a T helper 2 (Th2) phenotype more readily than did their Ndfip1(+/+) counterparts. This phenotype resembled that of Itchy mutant mice, suggesting that Ndfip1 might affect the function of Itch, an E3 ubiquitin ligase. We show that T cell activation promoted both Ndfip1 expression and its association with Itch. In the absence of Ndfip1, JunB half-life was prolonged after T cell activation. Thus, in the absence of Ndfip1, Itch is inactive and JunB accumulates. As a result, T cells produce Th2 cytokines and promote Th2-mediated inflammatory disease. PMID- 17137800 TI - The Integrin alpha9beta1 contributes to granulopoiesis by enhancing granulocyte colony-stimulating factor receptor signaling. AB - The integrin alpha9beta1 is widely expressed on neutrophils, smooth muscle, hepatocytes, endothelia, and some epithelia. We now show that mice lacking this integrin have a dramatic defect in neutrophil development, with decreased numbers of granulocyte precursors in bone marrow and impaired differentiation of bone marrow cells into granulocytes. In response to granulocyte colony-stimulating factor (G-CSF), alpha9-deficient bone marrow cells or human bone marrow cells incubated with alpha9beta1-blocking antibody demonstrated decreased phosphorylation of signal transducer and activator of transcription 3 and extracellular signal-regulated protein kinase. These effects depended on the alpha9 subunit cytoplasmic domain, which was required for formation of a physical complex between alpha9beta1 and ligated G-CSF receptor. Integrin alpha9beta1 was required for granulopoiesis and played a permissive role in the G-CSF-signaling pathway, suggesting that this integrin could play an important role in disorders of granulocyte development and other conditions characterized by defective G-CSF signaling. PMID- 17137801 TI - Various local hemostatic agents with different modes of action; an in vivo comparative randomized vascular surgical experimental study. AB - OBJECTIVES: To evaluate the effects of different local hemostatic agents in a new high flow vascular experimental bleeding model. DESIGN: Bovine thrombin combined with collagen matrix (bTcM), microporous polysaccharide hemospheres (MPH), freeze dried rFVIIa with and without the combination of MPH were compared to a control group (solely compression) in a randomized fashion (20 animals/group). Primary endpoint was hemostasis, and secondary endpoints were time to hemostasis, blood loss, and blood pressure at hemostasis. METHODS: The common carotid artery of heparinized rats was ligated proximally and transected. Compression was applied for one minute followed by application of the topical hemostatic agent. Compression was maintained for another two minutes followed by re-evaluation of hemostasis: if bleeding continued additional compression was applied and thereafter bleeding was checked every minute until hemostasis. RESULTS: All animals in the bTcM group obtained hemostasis compared to 20% in the control group (p<0.0001). The combination of MPH and rFVIIa (70% hemostasis) also showed a significant hemostatic capacity compared to control group (p<0.001). None of the other active treatment groups differed compared to control group. Animals treated with bTcM had a significantly shorter time to hemostasis compared to animals in the other active treatment groups. No significant difference in blood loss and blood pressure at hemostasis was detected. CONCLUSIONS: The most effective hemostatic agent was bTcM, followed by the combination of rFVIIa and MPH, while neither MPH nor rFVIIa alone displayed any hemostatic capacity compared to compression only. PMID- 17137802 TI - A novel approach to the management of the diabetic foot: metatarsal excision in the treatment of osteomyelitis. AB - OBJECTIVE: To describe the procedure and outcomes of metatarsal excision in seven patients treated for osteomyelitis in the diabetic foot. RESULTS: Average age was 60.6 (48-83) years. The mean length of hospital stay was 33.5 (3-50) days (excluding one patient who died from hospital acquired pneumonia). All remaining patients had negative wound cultures after a mean 7.4 (0-20) days of antibiotic treatment after procedure and were discharged from hospital 16.9 (2-48) days after surgery. Two patients developed wound infections after discharge. Pre operative levels of mobility were achieved within a mean of 12.6 days (range 2 40). CONCLUSIONS: In diabetic patients, metatarsal excision may be better than transmetatarsal amputation. PMID- 17137803 TI - Letter to editor re: endovascular repair of para-anastomotic aortic aueurysms. PMID- 17137804 TI - The high prevalence of colorectal neoplasms in preoperative patients with abdominal aortic aneurysm or peripheral artery disease. AB - OBJECTIVE: In Japan, the incidence of both colorectal carcinoma and vascular disease is increasing. We screened preoperative patients with abdominal aortic aneurysm (AAA) or peripheral artery disease (PAD) for colorectal cancer. DESIGN OF STUDY: This study was retrospective and cross-sectional. MATERIALS: The subjects were 492 patients admitted for elective surgery of AAA or PAD. METHODS: The patients underwent immunochemical faecal occult blood tests (FOBT) before operation, and those with positive results underwent investigations for colorectal neoplasm. We compared the results with that of screening programmes performed on the general population. RESULTS: Of the 408 patients that underwent FOBT, 104 (25.5%) were positive. After colonoscopy, six (1.5%) had colorectal carcinoma and 16 (3.9%) had advanced adenoma. These values were several folds higher than that for the general population in Japan. CONCLUSIONS: Patients with AAA or PAD carry a high risk for colorectal neoplasm. PMID- 17137805 TI - Relation between hospital volume and outcome of elective surgery for abdominal aortic aneurysm: a systematic review. AB - OBJECTIVES: Our aim was to analyse the relation between hospital volume and peri operative mortality in abdominal aortic aneurysm surgery. DESIGN: Systematic review. METHOD: The Medline, Embase and Cochrane databases were searched to identify all population based studies reporting on the volume outcome relationship published between 1966 and 2006. Two independent observers performed methodological quality assessment and data extraction. Outcome was 30-day or in hospital mortality in relation to hospital volume. RESULTS: Twenty-four articles were included. Overall peri-operative mortality ranged from 2.3 to 9.9%. The cut off values for a high- or low-volume hospital appeared to range from 8 to 50 operations annually. The peri-operative mortality in low volume hospitals (LVH) ranged from 3.0 to 13.8% (median 6.2%) and from 1.8 to 7.4% in high volume hospitals (HVH) (median 4.3%). In 14 studies a significantly lower mortality was found in HVH as opposed to LVH; in 10 articles no such difference between HVH and LVH could be proved. CONCLUSION: We found some evidence for a relation between the volume of AAA surgery and peri-operative mortality. There seems to be a nonsignificant trend in favour of high volume hospitals. However we could not derive an unequivocal volume threshold for safely performing AAA surgery. PMID- 17137806 TI - Intravascular stapler for "open" aortic surgery: preliminary results. AB - OBJECTIVES: The aim of this study was to assess the efficacy of a new stapling device using a pig model. METHODS: Straight 12 mm Gore-Tex grafts were inserted end to end into the aorta of 12 pigs. One anastomosis was performed with the stapler and the other using 4/0 prolene sutures and 13 mm needles. The animals were sacrificed at one week, one and three months and all grafts underwent histological examination. Leakage from the anastomoses was assessed in a separate specially designed circulation model using saline as a perfusate. RESULTS: The stapled anastomoses took 1.0+/-0.25 minutes to complete while suturing took 8.5+/ 1.5 minutes. There was no difference in the histology between the two types of anastomosis. The leak rate was six times greater at the sutured compared to the stapled anastomosis. CONCLUSION: The use of stapled anastomoses may allow a significant shortening of aortic cross clamping time, reduce anastomotic leakage and may be particularly useful in laparoscopic aortic repair. A randomised trial is required to assess the efficacy of this device. PMID- 17137808 TI - Plantar flexion as an alternative to treadmill exercise for evaluating patients with intermittent claudication. AB - OBJECTIVE: The aim of this study was to examine whether the plantar flexion test could adequately replace treadmill testing in patients who were unable to exercise. DESIGN: Prospective observational study. PATIENTS: Twenty-seven patients with intermittent claudication secondary to peripheral arterial disease (PAD). METHODS: Patients performed two treadmill tests and two plantar flexion tests. Ankle pressure, near infrared spectroscopy (NIRS) data, heart rate and blood pressures were monitored along with pain-free and maximum walking distances for treadmill, pain-free and maximum exercise time for plantar flexion. RESULTS: Maximum exercise time and walking distance were well correlated (R=0.74). Eleven patients (41%) developed non-claudicating symptoms during the treadmill test but not during the flexion test. Rate pressure product was significantly higher after the treadmill but not after the plantar flexion. CONCLUSIONS: Plantar flexion test showed good reliability and correlation. Plantar flexion may serve as an alternative to treadmill testing in evaluating muscle pain in patients with intermittent claudication. PMID- 17137810 TI - Cell signaling in lens development. PMID- 17137809 TI - Wall stress analysis in small asymptomatic, symptomatic and ruptured abdominal aortic aneurysms. AB - OBJECTIVES: To evaluate the potential of wall stress analysis for the identification of abdominal aortic aneurysm (AAA) at elevated risk of rupture in spite of small diameter. MATERIALS AND METHODS: Thirty patients with small AAA, 10 asymptomatic, 10 symptomatic and 10 ruptured, were included. Demographic data and results from physical examinations were recorded in a retrospective fashion. After CT-evaluation and the creation of a patient specific 3D model, wall stress was calculated using the finite element method. RESULTS: No differences were observed in diameter between asymptomatic, symptomatic or ruptured aneurysms (5.1+/-0.2 cm vs. 5.1+/-0.2 cm vs. 5.3+/-0.2 cm respectively; p=0.57). Peak aortic wall stress at maximal systolic blood pressure is significantly higher in ruptured than asymptomatic aneurysms (51.7+/-2.4 N/cm(2) vs. 39.7+/-3.3 N/cm(2) respectively; p=0.04). Wall stress analysis at uniform blood pressure, performed to correct for higher blood pressure in the symptomatic and rupture group did not result in significant differences in peak wall stress (asymptomatic 31.7+/-2.3 N/cm(2); symptomatic 30.5+/-1.3 N/cm(2); rupture 36.7+/-4.0 N/cm(2); p=0.26). CONCLUSIONS: Wall stress analysis at maximal systolic blood pressure is a promising technique to detect aneurysms at elevated aneurysm rupture risk. Since no significant differences were found at uniform blood pressure, the need for adequate blood pressure control in aneurysm patients is reiterated. PMID- 17137811 TI - Pain. PMID- 17137812 TI - Pattern recognition receptors in companion and farm animals - the key to unlocking the door to animal disease? AB - The innate immune system is essential for host defence and is responsible for early detection of potentially pathogenic microorganisms. Upon recognition of microbes by innate immune cells, such as macrophages and dendritic cells, diverse signalling pathways are activated that combine to define inflammatory responses that direct sterilisation of the threat and/or orchestrate development of the adaptive immune response. Innate immune signalling must be carefully controlled and regulation comes in part from interactions between activating and inhibiting signalling receptors. In recent years, an increasing number of pattern recognition receptors (PRRs), including C-type lectin receptors and Toll-like receptors (TLRs), has been described that participate in innate recognition of microbes, especially through the so called pathogen-associated molecular patterns (PAMPs). Recent studies demonstrate strong interactions between signalling through these receptors. Whereas useful models to study these receptors in great detail in the murine and human system are now emerging, relatively little is known regarding these receptors in companion and farm animals. In this review, current knowledge regarding these receptors in species of veterinary relevance is summarised. PMID- 17137813 TI - Neurologic and neurodevelopmental manifestations of pediatric HIV/AIDS: a global perspective. AB - Neurodevelopmental abnormalities associated with HIV infection have been described since the first reports of pediatric AIDS in the 1980s. Before antiretroviral therapy (ART) became widely available, progressive HIV-1 encephalopathy (PHE) was reported in the US in 13-35% of children with HIV-1 infection and in 35-50% of children with AIDS. Introduction of ART can prevent PHE and reverse PHE present at ART initiation, but a high prevalence of residual problems has been described. Even though 90% of HIV-infected children live in the developing world, few children have access to ART and little is known regarding the neurological manifestations of perinatal HIV infection in those regions. Mechanisms of pediatric HIV-1 neuropathogenesis and factors associated with neurodevelopmental abnormalities in perinatally infected children are not yet fully understood. Studies have demonstrated that HIV-1 enters the CNS soon after infection and may persist in this compartment over the entire course of HIV-1 infection. The CNS is a distinct viral reservoir, differing from peripheral compartments in target cells and antiretroviral penetration. Neurotropic HIV-1 likely develops distinct genotypic characteristics in response to this unique environment. We reviewed the literature on pediatric neuroAIDS and identified gaps in the current knowledge. PMID- 17137814 TI - Airway irritation and cough evoked by inhaled cigarette smoke: role of neuronal nicotinic acetylcholine receptors. AB - In a series of studies carried out in different experimental models, we investigated the type(s) of lung afferents and mechanism(s) underlying the cigarette smoke-induced airway irritation and cough. In healthy non-smokers, the intensity of airway irritation and cough evoked by cigarette smoke was markedly reduced after premedication with hexamethonium. A similar pattern of responses was also triggered by inhalation of nicotine aerosol. These studies in human subjects suggested nicotine as the primary causative agent in cigarette smoke that evokes airway irritation. Indeed, single-fiber recording experiments performed in anesthetized dogs showed that both C-fibers and rapidly adapting receptors in the lungs and airways were stimulated by inhalation of one puff of cigarette smoke, and the intensity of this stimulatory effect was related to the nicotine content in the cigarette and abolished by hexamethonium. To further study the direct effect of nicotine on these sensory nerves, we measured the change in intracellular calcium concentration ([Ca(2+)](i)) of pulmonary sensory neurons isolated from the nodose and jugular ganglia of adult rats. Our results showed that nicotine evoked an abrupt and transient increase in [Ca(2+)](i) in approximately 34% of the 522 neurons tested, and 1,1-dimethyl-4 phenylpiperazinium, a selective agonist of the neuronal nicotinic acetylcholine receptors (NnAChRs), evoked a similar pattern of response as that of nicotine in these neurons. In conclusion, results of these studies show that nicotine exerts a direct stimulatory effect on vagal pulmonary sensory neurons. This stimulatory effect of nicotine is primarily responsible for the airway irritation and cough evoked by inhaled cigarette smoke, and is mediated through an activation of the NnAChRs. PMID- 17137815 TI - Sodium-dependent phosphate transport across the apical membrane of alveolar epithelium in caprine mammary gland. AB - NaPi IIb cotransporter is expressed in various tissues including mammary glands of mice. The physiological role of NaPi IIb in lactating mammary glands is still unclear. Therefore, it was the aim of the study to detect and to localize NaPi IIb protein in lactating goat mammary glands by Western analysis and immunohistochemistry. Furthermore, Na(+)-dependent P(i) uptake into apical membrane vesicles isolated from goat milk was determined using rapid filtration technique. NaPi IIb protein could specifically be detected in the apical membranes of lactating alveolar epithelial cells. Na(+)-dependent P(i) uptake into apical membrane vesicles could be measured, which was inhibited by phosphonoformic acid. The kinetic parameters were V(max) with 0.9 nmol/mg protein/10 s and K(m) with 0.22 mmol/L for P(i) affinity, K(m) value for Na(+) affinity 11 mmol/L. Stoichiometry of this mammary gland Na(+)/P(i) transport across the apical membranes seemed to be 1:1 P(i):Na(+) without cooperativity in P(i) and Na(+) binding as assessed by Scatchard and Hill plots. These features of Na(+)/P(i) transport suggest that it could be mediated by NaPi IIb. The quantitative role of this P(i) transport which is directed from the alveolar lumen into the epithelial cell of goat mammary gland will be the topic of further investigations. PMID- 17137816 TI - Contractile properties of the functionally divided python heart: two sides of the same matter. AB - The heart of Python regius is functionally divided so that systemic blood pressure is much higher than pulmonary pressure (6.6+/-1.0 and 0.7+/-0.1 kPa, respectively). The present study shows that force production of cardiac strips from the cavum arteriosum and cavum pulmonale exhibits similar force production when stimulated in vitro. The high systemic blood pressure is caused, therefore, by a thicker ventricular wall surrounding the cavum arteriosum rather than differences in the intrinsic properties of the cardiac tissues. Similarly, there were no differences between the contractile properties of right and left atria. Force production was similar in atria and ventricle but the atria contracted and relaxed much faster than the ventricle. Graded hypoxia markedly reduced twitch force of all four cardiac tissues, and this was most pronounced when PO(2) was below 40 kPa. In contrast, the four cardiac tissues were insensitive to acidosis during normoxia although acidosis increased the sensitivity to hypoxia. Adrenergic stimulation increased twitch force of all cardiac tissues, while cholinergic stimulation only affected the atria and reduced twitch force markedly. In spite of the different oxygen availability of the two sides of the heart, the biochemical and functional properties are alike and the differences may instead be overcome by the coronary blood supply. PMID- 17137817 TI - Dermatan sulfate is the major metachromatic glycosaminoglycan in the integument of the anuran Bufo ictericus. AB - Glycosaminoglycans from the ventral and dorsal integuments of the anuran Bufo ictericus were characterized based on biochemical and histochemical methods. Dermatan sulfate is the major metachromatic glycosaminoglycan found in these tissues, but small amount of heparan sulfate was also detected. The average molecular mass of the dermatan sulfate is approximately 20 kDa, similar to the glycosaminoglycan isolated from mammalian skin. In addition, the amphibian integument contains high amounts of hyaluronic acid, especially in the ventral area. We also observed that the glycosaminoglycans occur in the anuran integument as irregular deposits through the spongious dermis and in the mast cells, as revealed by histochemical analysis using Alcian blue, dimethylmethylene blue and toluidine blue stains. The concentration and composition of glycosaminoglycans found in the amphibian integument resemble those from mammalian skin except for the higher concentration of hyaluronic acid in the amphibian tissue. Possibly, this observation indicates that the function of the sulfated glycosaminoglycan in these tissues has been preserved during evolution, although the amphibian integument and the human skin have their own particular physiology. PMID- 17137818 TI - [Primary non-Hodgkin lymphoma of the liver: case report and review of the literature]. AB - Purpose was to describe the clinical, radiological and therapeutic features in primary liver lymphoma. We report the case of a 54-year-old patient, who is followed since the age of 20 years for neutropenia associated with mediastinal adenopathy. Systematical ultrasound find a mass of the left liver confirmed by Computed tomography (CT). Histological examination of laparoscopic liver biopsy specimens confirmed diffuse large-cell non-Hodgkin's lymphoma. The disease was confined to the liver without any evidence of extrahepatic involvement. The serology of Epstein Barr virus was highly positive. PET-scan show increased FDG uptake at the site of hepatic lesion and the mediastin. The patient received chemotherapy followed by radiation therapy of the left liver at the dose of 31 Gy. The patient was alive and free of disease 20 months after the diagnosis of primary liver lymphoma. The primary hepatic lymphoma is a rare malignancy, which classically affects 50-year-old patients with a male preponderance. The incidence is increased in immunosuppressed patients and some authors have suggested an association with hepatitis B or C infection, and with the Epstein Barr virus. The imaging studies including ultrasound, CT, magnetic resonance imaging (MRI) and now PET-scan help to establish the diagnosis and to the following. Treatment options are surgery, radiation, chemotherapy, or a combination. PMID- 17137819 TI - [Positron emission tomography using 18F-fluorodeoxyglucose in the management of nonsmall cell lung cancers in 2006]. AB - Technological progress and numerous published studies allow to estimate the best place of the 18F-fluorodeoxyglucose positron emission tomography, a real functional metabolic imagery, in the clinical and therapeutic strategy of non small cell lung cancers. PMID- 17137820 TI - Hip pain from impingement and dysplasia in patients aged 20-50 years. Workup and role for reconstruction. AB - In the 20-50-year age group, hip pain usually indicates dysplasia. Chronic mechanical pain is the usual pattern, although acute pain caused by avulsion or degeneration of the labrum may occur. The morphological characteristics of the dysplastic hip should be evaluated, and the link between the dysplasia and the osteoarthritis should be confirmed. Three factors indicate a favorable prognosis: joint space preservation, age younger than 40 years, and correctable femoral and acetabular abnormalities. Reconstruction is highly desirable, as it delays the need for joint replacement by 20 years. After 15 years, good outcomes are seen in 87% of patients after shelf arthroplasty and 85% after femoral varus osteotomy with or without shelf arthroplasty. Chiari acetabular osteotomy can be performed in patients with osteoarthritis but is followed by prolonged limping. Periacetabular osteotomy should be reserved for patients with moderate dysplasia and no evidence of osteoarthritis. Shelf arthroplasty and femoral osteotomy require 5-8 months off work (compared to 5 months after hip replacement surgery) but subsequently permits a far more active lifestyle. Hip replacement, which is required 20 years or more after biologic reconstruction, carries the same prognosis as first-line hip replacement (good results in 80% of patients after 15 years). Acute sharp pain related to anterior hip derangement also occurs in primary femoroacetabular impingement (FAI). The most common pattern is cam impingement, which is due to a decrease in head-neck offset and manifests as pain during flexion and adduction of the hip. Cam impingement can be corrected by anterolateral osteoplasty, which is often performed arthroscopically. Pincer-type impingement is contact between the anterior acetabular rim and the femoral neck due to retroversion of the proximal acetabulum. The imaging study strategy is discussed. Coxometry, computed tomography, and arthrography can be used. Primary FAI, which occurs as a result of geometric abnormalities, should be distinguished from secondary impingement. Causes of secondary impingement include exaggerated lumbar lordosis with pelvic tilt and to hip osteophytosis (sports or posterior hip osteoarthritis). Osteoplasty is rarely appropriate in patients with secondary impingement. The features of acute anterior hip derangement are now better defined. They can be used to guide palliative treatment, which is effective, in the medium term at least. Experience acquired over the last two decades has established the efficacy of surgery for hip dysplasia. PMID- 17137821 TI - [Results of surgical clitoral repair after ritual excision: 453 cases]. AB - OBJECTIVE: Ritual excision is responsible for urologic, gynaecologic and obstetrical complications, whose surgical treatment has been fully described. Sexual sequelae deserve the same attention. We describe and analyze the results of a surgical procedure for clitoral rehabilitation. PATIENTS AND METHODS: Women requesting this surgery between 1992 and 2005 have been prospectively included in this study. The skin covering the stump was resected and the clitoris identified. The suspensor ligament was sectioned in order to mobilize the stump, the sclerous tissues were removed from the extremity and the neo glans brought to a normal situation. Pre operative pain and clitoral impairment were assessed within five categories. The same was done with anatomical and functional postoperative results at six months. RESULTS: Four hundred and fifty-three patients have been completely evaluated. Hospitalisation stay was 24 hours and the procedure never lasted more than 30 minutes. Minor early complications were recorded in 58 patients (hematoma, disrupture of the suture, pain). Four patients reported persisting pain at four months. A visible clitoral massif could be restored in 87% of the cases and a real improvement in clitoral function was obtained in 75% of the patients. DISCUSSION AND CONCLUSION: This surgical procedure is easy and reliable. It provides promising cosmetic and functional results with minor complications. PMID- 17137822 TI - [Patient information booklet SOR SAVOIR PATIENT: understanding ovarian cancer, a guide for cancer patients and family members]. PMID- 17137823 TI - X-ray micro-tomography of a coronary stent deployed in a model artery. AB - Coronary stents are small scaffolds routinely implanted during angioplasty procedures to re-open coronary arteries which have become narrowed by an atherosclerotic plaque. Despite the advent of drug-eluting stents to reduce restenosis (re-narrowing) after the procedure, complications still arise and mechanical factors may be partly the cause. Stents are being used in increasingly complex cases, so new forms of pre-clinical testing may be helpful in evaluating stent designs and deployment techniques. With this in mind, an in vitro experiment was conducted to evaluate the use of X-ray micro-tomography to image stents at various stages of deployment. A stent was deployed in an artificial artery while using a synchrotron X-ray source to obtain the tomography scans. A volume element (voxel) size of 5.3 microm was achieved, with a vertical field of view of 4.5mm. The imaging of the stent and artificial artery materials was better than expected, despite some attenuation artifacts and the high monochromatic beam energy used (25 keV). Experimental problems are discussed, together with recommendations for improving the technique. It is intended that this technique will be of interest to engineers and clinicians as a pre-clinical test. PMID- 17137824 TI - An investigation of leakage tracts along stressed suture lines in phantom tissue. AB - Suturing is still one of the most important procedures used in trauma and surgery environments. It does however rely on piercing other wise healthy tissue. In doing so channels are opened which may act as conduits for bacteria to enter the body or to allow material such as blood or intestinal bacteria to leak from vessels. This work shows by means of finite element analysis and physical modelling through the use of a phantom that the tension in the suture has a direct bearing on the opening of these channels. It was found that for the phantom the channel cross-sectional area was approximately proportional to the applied tension and suture thread deflection. Leakage channels of up to 0.1mm(2) cross-section were achieved with suture tensions of 120 grammes force. Channels of this size are significantly larger than the bacteria suggesting transmission is theoretically possible. PMID- 17137825 TI - Analytical analysis of the Pennes bioheat transfer equation with sinusoidal heat flux condition on skin surface. AB - This study focuses on the effect of the temperature response of a semi-infinite biological tissue due to a sinusoidal heat flux at the skin. The Pennes bioheat transfer equation such as rho(t)c(t)( partial differentialT/ partial differentialt)+W(b)c(b)(T-T(a))=k partial differential(2)T/ partial differentialx(2) with the oscillatory heat flux boundary condition such as q(0,t)=q(0)e(iomegat) was investigated. By using the Laplace transform, the analytical solution of the Pennes bioheat transfer equation with surface sinusoidal heating condition is found. This analytical expression is suitable for describing the transient temperature response of tissue for the whole time domain from the starting periodic oscillation to the final steady periodic oscillation. The results show that the temperature oscillation due to the sinusoidal heating on the skin surface is unstable in the initial period. Further, it is unavailable to predict the blood perfusion rate via the phase shifting between the surface heat flux and the surface temperature. Moreover, the lower frequency of sinusoidal heat flux on the skin surface induces a more sensitive phase shift response to the blood perfusion rate change, but extends the beginning time of sampling because of the avoidance of the unavailable first cyclic oscillation. PMID- 17137826 TI - Investigations into the relationship between hemodynamics and vascular alterations in an established arteriovenous fistula. AB - Arteriovenous fistula are specific vessels created by a vascular operation in order to provide sufficient blood access for extracorporeal circulation in hemodialysis. They are subject to numerous pathologies that may be caused by hemodynamic effects. To better understand these effects, a specific patient's arteriovenous fistula was reconstructed from computed tomography angiography. Computational fluid dynamics software made it possible to solve fluid mechanics equations under physiological conditions. An accurate map of unsteady velocity profiles and wall shear stress was drawn up. The computed velocity profiles were successfully confronted with Echo Doppler investigation. Selected regions with or without calcification, the end stage of wall alteration, were examined in terms of the mechanical constraints generated by blood flow. In contrast with other authors, we did not observe any association between calcification and areas of oscillating shear stress. Nevertheless, a statistical analysis of the whole vessel envelop and specific sites of calcification suggested a potential association between calcification and high temporal wall shear stress gradients. PMID- 17137827 TI - Neogenin: one receptor, many functions. AB - Neogenin is a multifunctional transmembrane receptor belonging to the immunoglobulin superfamily. It displays identical secondary structure to deleted in colorectal cancer (DCC), a netrin receptor that is involved in axon guidance and cell survival. Like DCC, neogenin is able to transduce signals elicited by netrin. These neogenin-netrin interactions have been implicated in tissue morphogenesis, angiogenesis, myoblast differentiation and most recently in axon guidance. Neogenin is also a receptor for repulsive guidance molecule, a glycosylphosphatidylinositol-linked protein involved in neuronal differentiation, apoptosis and repulsive axon guidance. Numerous studies have been started to elucidate the in vivo functions of neogenin, and its role in multiple aspects of development and homeostasis. PMID- 17137828 TI - Duchenne muscular dystrophy: focus on pharmaceutical and nutritional interventions. AB - Duchenne muscular dystrophy is a lethal X-linked muscle disease resulting from a defect in the muscle membrane protein dystrophin. The absence of dystrophin leads to muscle membrane fragility, muscle death (necrosis) and eventual replacement of skeletal muscle by fat and fibrous connective tissue. Extensive muscle wasting and respiratory failure results in premature death often by the early 20s. This short review evaluates drug and nutritional interventions designed to reduce the severity of muscular dystrophy, while awaiting the outcome of research into therapies to correct the fundamental gene defect. Combinations of dietary supplementation with amino-acids such as creatine, specific anti-inflammatory drugs and perhaps drugs that target ion channels might have immediate realistic clinical benefits although rigorous research is required to determine optimal combinations of such interventions. PMID- 17137829 TI - Beyond perception: synaesthesia as a psycholinguistic phenomenon. AB - Synaesthesia has been described as a perceptual phenomenon that creates a 'merging of senses'. Therefore, academic treatments have focused primarily on its sensory characteristics and similarities with veridical perception. This approach has dominated, despite parallel work that has suggested conceptual influences are involved, including data that show a large number of synaesthetic variants are triggered by linguistic symbols (e.g. words). These variants are the focus of a novel subfield that applies psycholinguistic methodology to the study of linguistic synaesthesias. This approach is redefining notions of synaesthesia and of the interplay between perceptual and non-perceptual systems, in addition to informing general theories of language. This review examines the emergent field of linguistic synaesthesia research and the broad range of linguistic mechanisms that are implicated. PMID- 17137830 TI - Summary of major conclusions from the 4th IWGT, San Francisco, 9-10 September, 2005. PMID- 17137831 TI - Enhancement of phase II enzyme activity by purpurin resulting in the suppression of MeIQx-DNA-adduct formation in mice. AB - We previously demonstrated using a bacterial system that the antigenotoxic activity of the anthraquinone compounds purpurin and alizarin was due to the suppression of microsomal enzyme activity involved in the activation of mutagens. In the present study we determined the effect of purpurin and alizarin on (i) MeIQx-DNA-adduct formation in mouse tissues and (ii) the activity of phases I and II enzymes in liver fractions, the liver being the target tissue of MeIQx. The amount of MeIQx-DNA adduct formed was determined using 32P-postlabeling methods. Methoxyresorufin-O-demethylase (MROD) and ethoxyresorufin-O-deethylase (EROD) enzyme activities, which reflect CYP 1A activity, were measured as markers for phase I enzymes, and UDP-glucuronyltransferase (UGT) and glutathione S transferase (GST) activities were determined as markers for phase II enzymes. Mice fed with a diet containing 0.5% purpurin for 3 days prior to MeIQx administration had 70% fewer MeIQx-DNA adducts in the lung and kidney, and fewer DNA adducts (insignificant, statistically) in the liver compared with mice fed a diet lacking purpurin. MROD and EROD activities in the liver of these mice increased six- and eight-fold, respectively, and were higher than those determined for the control mice within 1 day following commencement of purpurin treatment. These elevated activities were maintained during treatment and declined immediately following removal of purpurin from the diet. GST and UGT activities gradually increased 2.5- and 3-fold, respectively, following purpurin treatment, and were maintained at significantly high levels even after purpurin administration ceased. Alizarin did not significantly affect DNA-adduct formation and enzyme activity, except in the case of UGT. Taken together, our results show that purpurin reduced MeIQx-DNA-adduct formation by maintaining elevated phase II enzyme activities, thereby facilitating accelerated excretion of MeIQx. PMID- 17137832 TI - Personal reflections on the life and legacy of Alexander Hollaender. AB - Mary Esther Gaulden presents a personal summary of the activities of Alexander Hollaender, from his days at the National Institutes of Health to his becoming Director of the Biology Division of the Oak Ridge National Laboratory in 1947. This appealing story deals with many of her reactions to his personality and organizational style. It reflects the atmosphere of science in those days, and her enthusiasm in this vibrant milieu. Next is a brief account by John Jagger of his first meeting with Dr. Hollaender, arrival in Oak Ridge in April 1956, and wedding to Mary Esther six months later at the house of the Hollaenders in Oak Ridge. The third section is an account by Virginia P. White of how she came to Oak Ridge in 1955 and became Dr. Hollaender's Laboratory Administrator. She gives a personal account of the many facets of his managerial style, as well as of the personality of his wife, Henrietta. She also describes one of Hollaender's many avocations, the collection of fossils on Sunday morning hikes in the Cumberland Mountains, accompanied by lab and visiting personnel, and finally comments on the annual research conferences in Gatlinburg TN, for which Hollaender and the lab became very well known, with some closing vignettes on his leadership style. PMID- 17137833 TI - For what purpose and reasons do doctors use the Internet: a systematic review. AB - OBJECTIVES: To determine doctors' reasons for using the Internet, and the factors that influence their usage. DATA SOURCES: A systematic review of 38 studies, from 1994 to 2004, describing surveys of doctors' Internet usage. RESULTS: All of the studies were in the developed world, primarily in North America. Approximately 60 70% of doctors have access to the Internet, but in several studies access is more than 90%. Access is steadily increasing. Most Internet activity focuses on email and searching in journals and databases, but there is a very wide range of activities. Professional email with colleagues and patients is low, but increasing. The major factors discouraging usage are time, workload and cost, while too much information, liability issues and lack of skills also feature as discouraging factors. Factors encouraging use are unclear, but overall patient satisfaction and belief in improved service delivery, time saving and demand from patients are factors. There is a trend that males use the Internet more than females, young more than old, and specialists more than generalists, but these differences are not across the board, and show variations between studies. CONCLUSION: In spite of the limitations, it is clear that doctors are highly connected to the Internet, and their professional usage is increasing. Factors encouraging and discouraging usage are more complex than simple connectivity. Usage differences between demographic groups do exist, but are equalising. More and consistent research is required in this area. PMID- 17137834 TI - Post-infectious fatigue syndrome in dengue infection. AB - BACKGROUND: Although the acute manifestations of dengue are well known, few studies have assessed the long-term consequences of dengue infection. We prospectively studied the incidence and factors associated with fatigue in a cohort of patients following dengue infection. METHODS: We included patients with serologically confirmed dengue infection admitted to the National University Hospital, Singapore, during a dengue outbreak from October-November 2005. The severity of dengue was graded as dengue fever, dengue haemorrhagic fever and dengue shock syndrome. A follow-up telephone interview was performed two months following hospital discharge, where a Fatigue Questionnaire was administered. The presence of significant fatigue was considered as the main outcome measure. Significance was assessed at P<0.05. RESULTS: One hundred twenty-seven patients, 71 (55.9%) males and 56 (44.1%) females, of mean age 36.06 years (range, 16-70; S.D., 13.722), participated in this study. Twenty-five (19.7%) patients had dengue haemorrhagic fever and the remaining 102 (80.3%) had dengue fever. In multivariate analysis, increased age, female sex, the presence of chills, and the absence of rashes were significantly associated with the development of fatigue post-dengue infection. There was no significant association between fatigue and dengue severity. CONCLUSIONS: This observation represents the first systematic evidence that dengue can result in clinical disease beyond the acute phases of infection. Host factors, such as age and sex may be important in the pathogenesis of this clinical entity. PMID- 17137835 TI - Human Coronavirus-NL63 infections in Korean children, 2004-2006. AB - BACKGROUND: Human coronavirus-NL63 (HCoV-NL63) has been isolated from children with respiratory tract infections and its prevalence in Korea has not been reported. OBJECTIVES: This study was designed to investigate the presence and the clinical features of HCoV-NL63 during two winter seasons. STUDY DESIGN: During April 2004-April 2006, nasopharyngeal specimens from children hospitalized with acute respiratory disease were tested for common respiratory viruses, including RSV, influenza A, influenza B, parainfluenza viruses, and adenovirus by IFA. hMPV infection was excluded by nested RT-PCR using primers for F-gene. To detect HCoV NL63, previously described nested PCR assays for 1a and 1b were used. PCR products of the 1a gene for HCoV-NL63 were sequenced. RESULTS: Out of 872 nasopharyngeal aspirate from children aged under 16 years, 14 (1.7%) were positive for HCoV-NL63. Most of the patients had croup (64.2%) or bronchiolitis (21.4%). The peak prevalence was found in November (28.5%). Most were collected between November 2004 and February 2005. CONCLUSIONS: HCoV-NL63 may be one of the causative agents of acute respiratory tract infection, especially croup. PMID- 17137836 TI - Cervical spine manipulation alters sensorimotor integration: a somatosensory evoked potential study. AB - OBJECTIVE: To study the immediate sensorimotor neurophysiological effects of cervical spine manipulation using somatosensory evoked potentials (SEPs). METHODS: Twelve subjects with a history of reoccurring neck stiffness and/or neck pain, but no acute symptoms at the time of the study were invited to participate in the study. An additional twelve subjects participated in a passive head movement control experiment. Spinal (N11, N13) brainstem (P14) and cortical (N20, N30) SEPs to median nerve stimulation were recorded before and for 30min after a single session of cervical spine manipulation, or passive head movement. RESULTS: There was a significant decrease in the amplitude of parietal N20 and frontal N30 SEP components following the single session of cervical spine manipulation compared to pre-manipulation baseline values. These changes lasted on average 20min following the manipulation intervention. No changes were observed in the passive head movement control condition. CONCLUSIONS: Spinal manipulation of dysfunctional cervical joints can lead to transient cortical plastic changes, as demonstrated by attenuation of cortical somatosensory evoked responses. SIGNIFICANCE: This study suggests that cervical spine manipulation may alter cortical somatosensory processing and sensorimotor integration. These findings may help to elucidate the mechanisms responsible for the effective relief of pain and restoration of functional ability documented following spinal manipulation treatment. PMID- 17137837 TI - Upper motor neuron involvement in X-linked recessive bulbospinal muscular atrophy. AB - OBJECTIVE: Clinicopathological findings of X-linked recessive bulbospinal muscular atrophy (SBMA) are indicative of lower motor neuron and primary sensory neuron involvement. The aim of our study was to investigate the presence of subclinical upper motor neuron (UMN) dysfunction in this disease. METHODS: Two siblings with clinical presentation, routine electrophysiological tests, histopathological features of muscle and nerve biopsies and genetic testing consistent with diagnosis of SBMA underwent transcranial magnetic stimulation (TMS). The analysed parameters were motor evoked potential (MEP) threshold, silent period (SP) and central motor conduction time. Intracortical inhibition with paired pulses from 1 to 6ms interstimulus intervals (ISIs) was evaluated in the older brother. RESULTS: MEP parameters were significantly altered in limb and cranial muscles and MEP suppression after paired stimulation significantly reduced in the older brother. MEP abnormalities were present in one lower limb, but SP abolished in all limbs, in the younger brother. CONCLUSIONS: Subclinical involvement of UMNs may be present in patients affected by SBMA. This finding suggests that the array of neuronal systems whose function may be affected by the pathogenic process of SBMA is larger than it was considered so far. SIGNIFICANCE: TMS is a sensitive diagnostic tool for the identification of UMN dysfunction and should be included in the diagnostic evaluation of patients with SBMA. PMID- 17137838 TI - Obstructive sleep apnoea and adipocyte death. PMID- 17137839 TI - Polyclonal antibodies mediated immobilization of a peroxidase from ammonium sulphate fractionated bitter gourd (Momordica charantia) proteins. AB - Polyclonal antibody bound Sepharose 4B support has been exploited for the immobilization of bitter gourd peroxidase directly from ammonium sulphate precipitated proteins. Immunoaffinity immobilized bitter gourd peroxidase exhibited high yield of immobilization. IgG-Sepharose 4B bound bitter gourd peroxidase showed a higher stability against heat, chaotropic agents (urea and guanidinium chloride), detergents (cetyl trimethyl ammonium bromide and Surf Excel), proteolytic enzyme (trypsin) and water-miscible organic solvents (propanol, THF and dioxane). The activity of immobilized bitter gourd peroxidase was significantly enhanced in the presence of cetyl trimethyl ammonium bromide and after treatment with trypsin as compared to soluble enzyme. PMID- 17137840 TI - Genospecies of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks from the Autonomous Province of Trento, Italy. AB - Sequences of the variable intergenic spacer region 5S (rrfA) 23S (rrlB) rRNA were used to identify Borrelia genospecies present in Ixodes ricinus nymphs collected from the Lamar Lakes area of the Province of Trento, Italy (overall prevalence=6.3%). Four genospecies were identified, one for the first time in this Province (B. valaisiana), and three which have been noted previously (B. afzelii, B. garinii, and B. burgdorferi s.s.). In order to compare the genetic variability of these genospecies in Trento with that at a European level, our 21 sequences (15 new haplotypes) and all appropriate European Borrelia sequences registered in GenBank (up to the end of 2004) were subjected to a phylogenetic analysis (for a total of 73 sequences and 43 haplotypes). Clusters of sequences representing the five main European genospecies (afzelii, garinii, burgdorferi s.s., valaisiana, lusitaniae) are well-supported. At least two other groups of haplotypes (genospecies) are suggested by our analysis; moreover, divergent evolution may be occurring in several genospecies. The maximum uncorrected pairwise differences between sequences within genospecies ranges from 1.5% (B. burgdorferi s.s.), to 2.3% (B. garinii and B. valaisiana) to 4.7% (B. afzelii), and are not correlated with geographical distribution. Within the Province of Trento, these values for the same genospecies are 1.5%, 2.3%, 0.9%, 1.9%, respectively. These high mutation rates within genospecies suggest that the sequencing of haplotypes should continue if we are to fully understand and monitor the evolution and epidemiology of Borrelia. PMID- 17137841 TI - Anti-IgA antibodies in common variable immunodeficiency (CVID): diagnostic workup and therapeutic strategy. AB - Common Variable Immunodeficiency (CVID) patients who are seropositive for anti IgA antibodies have a predisposition for anaphylactoid reactions to intravenous immunoglobulin replacement therapy (IVIG). Among 88 CVID patients, we identified eight with IgG anti-IgA antibodies (9%). All eight completely lacked IgA (<0.0009 g/l). Five of them had a history of anaphylactoid reactions to IVIG. However, four of these five patients tolerated subcutaneous immunoglobulin replacement therapy (SCIG). To identify predisposing factors for anti-IgA antibodies and related anaphylactoid reactions, we analyzed the clinical and immunological phenotype of affected patients. All eight IgG anti-IgA-positive patients lacked IgA(+) B cells in peripheral blood. Moreover, CVID patients with retained class switched CD27(pos) IgM(neg) IgD(neg) memory B cells (Freiburg classification group II) and total IgA deficiency seem to have an increased risk for developing anti-IgA antibodies. In seven of the eight patients, lymphoproliferation was observed (most prominently nodular lymphatic hyperplasia), two had granulomatous disease, and two showed autoimmune phenomena. PMID- 17137842 TI - Contrast-enhanced tissue Doppler imaging of the left atrial appendage is a new quantitative measure of spontaneous echocardiographic contrast in atrial fibrillation. AB - AIMS: Although left atrial appendage spontaneous echo contrast (LAASEC) is a marker of increased thromboembolic risk in atrial fibrillation, it has previously only been evaluated qualitatively. We sought to determine if an intravenous contrast echocardiographic agent combined with tissue Doppler imaging (TDI) of the LAA could accurately quantify LAA-SEC in patients with atrial fibrillation. METHODS AND RESULTS: We prospectively identified 55 patients with persistent atrial arrhythmias (mean age 63+/-13 years) undergoing a transesophageal echocardiography (TEE), with LAA-SEC prior to direct current cardioversion. In addition to off-line calculation of backscatter index and shear rate, quantification of the velocity in a color TDI region of interest was performed in the LAA cavity following a 0.5-mL intravenous bolus of Optison. LAA-SEC was qualitatively graded by a blindedreader as mild (n = 29) or severe (n = 26), and was compared off-line to TEE-derived quantitative variables. Compared to patients with mild LAA-SEC, those with severe LAA-SEC had significantly decreased LAA emptying velocity, LAA TDI mean velocities and shear rate. Over the whole group, the mean maximal velocity of the LAA using TDI correlated with LAA emptying velocity (r = 0.59; P < 0.0001), shear rate (r = 0.55; P < 0.0001) and LAA area (r = 0.34; P = 0.014). Severe LAA-SEC was found with 72% sensitivity and 82% specificity if TDI mean velocity was <6.13 cm/s. On logistic regression analysis, LAA-TDI was the only predictor of qualitative LAA-SEC grade. CONCLUSION: Contrast enhanced TDI is an original new tool that provides a quantification of the mean velocity of LAA-SEC that might improve our decision making in patients with atrial fibrillation. PMID- 17137843 TI - Fatty acid composition and lipid peroxidation of soft-shelled turtle, Pelodiscus sinensis, fed different dietary lipid sources. AB - Juvenile soft-shelled turtles (Pelodiscus sinensis) were fed 7 diets containing 8% of lard, soybean oil, olive oil, menhaden fish oil, or mixtures of 1 to 1 ratio of fish oil and lard, soybean oil, olive oil for 10 weeks. Growth and muscle proximate compositions of the turtles were not affected by different dietary treatments (p>0.05). Fatty acid profiles in muscle polar lipids, muscle non-polar lipids, and liver polar lipids reflected the fatty acid composition of dietary lipid source. Turtles fed diets containing fish oil generally contained significantly higher (p<0.05) proportion of highly unsaturated fatty acids (HUFA) in both polar and non-polar lipids of muscle and polar fraction of liver lipids than those fed other oils. Non-polar fraction of liver lipids from all groups of turtles contained less than 1% of HUFA. All turtles contained relatively high proportions of oleic acid in their lipids regardless of the dietary lipid source. Further, lipid peroxidation in both muscle tissue and liver microsomes of turtles fed fish oil as the sole lipid source was greater (p<0.05) than those fed fish oil-free diets. Turtles fed olive oil as the sole lipid source had the lowest lipid peroxidation rate among all dietary groups. The results indicate that dietary n-3 HUFA may not be crucial for optimal growth of soft-shelled turtles although they may be used for metabolic purpose. Further, high level of dietary HUFA not only increases the HUFA content in turtle tissues, but also enhances the susceptibility of these tissues to lipid peroxidation. PMID- 17137844 TI - The challenge of recruiting patients with anterior cruciate ligament injury of the knee into a randomized clinical trial comparing surgical and non-surgical treatment. AB - AIMS: To determine the number of patients needed to be screened (NNS) and allocated (NNA) in order to include one participant in a randomized clinical trial (RCT), and to compare the characteristics of patients accepting or declining participation in the RCT. METHODS: The recruitment process of an ongoing multicenter RCT, comparing surgical and non-surgical interventions after acute anterior cruciate ligament (ACL) injury of the knee is described. We use the known concept Number Needed to Screen (NNS) and introduce the new concept Number Needed to Allocate (NNA) as variables to support a priori sample size calculations of future investigations. RESULTS: 560 patients were screened to identify 162 patients (29%) eligible for inclusion in the RCT. 41 of those declined participation for various reasons, the most common being unwillingness to undergo surgery (n=23) or unwillingness to risk conservative treatment (n=8). 19 patients were excluded after MRI assessment or arthroscopy. Thus, 102 (18%) patients were allocated to one of the two treatments in the RCT. The NNS was 5.5 individuals with an acute knee injury, and the NNA was 1.6 individuals eligible for inclusion, to include 1 patient in the RCT. Patients declining to participate in the RCT were more frequently self-employed and less frequently injured in sports activities than those accepting RCT participation. CONCLUSIONS: We suggest that the a priori sample size calculation needs to be multiplied by at least 5.5 to provide an estimate of the number needed to screen, or 1.6 to provide an estimate of the number needed to allocate in order to include the desired number of patients in a trial comparing surgical and non-surgical treatment of the ACL injured patient. PMID- 17137845 TI - The hepatitis C virus epidemic in Cameroon: genetic evidence for rapid transmission between 1920 and 1960. AB - Hepatitis C virus (HCV) infection in Cameroon is characterized by widespread seropositivity and great virus genetic diversity (3 genotypes and over 10 subtypes). A total of 244 HCV NS5B sequences of 382-405 bp long (95 type 1, 58 type 2, and 91 type 4) were phylogenetically analyzed to estimate the history of the HCV epidemic in Cameroon. The newly developed Bayesian coalescent approach was used to infer the history of each HCV type. The estimated dates of the most recent common ancestors (MRCA) for genotypes 1 (1500; 95% confidence interval (95% CI): 1300-1650) and 4 (1500; 95% CI: 1350-1700) were in the same range, while the date for genotype 2 MRCA (1600; 95% CI: 1400-1750) was slightly more recent. The mean genetic distance between HCV genotype 1 sequences was greater than that of HCV type 4 sequences, itself greater than that of HCV type 2 sequences. The initial infected populations of all three genotypes did not grow until recently, when they grew exponentially. The growth rate has now begun to slow, with a less steep exponential growth curve. The period of exponential growth of all the three genotypes was between 1920 and 1960. These results (i) confirm that HCV genotypes 1 and 4 have produced long-term endemics, (ii) suggest that genotype 2 was introduced into Cameroon more recently, and (iii) indicate that the exponential spread of the three genotypes between 1920 and 1960 coincided with the mass campaign against trypanosomiasis and mass vaccinations in Cameroon. PMID- 17137846 TI - Burkholderia gladioli: five year experience in a cystic fibrosis and lung transplantation center. AB - BACKGROUND: The impact of infection with Burkholderia gladioli in cystic fibrosis, other chronic airway diseases and immunosuppressed patients is unknown. METHODS: A six-year retrospective review of all patients with B. gladioli infection was performed in a tertiary referral center with cystic fibrosis and lung transplantation programs. In addition, a targeted survey of all 251 lung transplant recipients was performed. Available B. gladioli isolates were analyzed via pulsed field gel electrophoresis. RESULTS: Thirty-five patients were culture positive for B. gladioli, including 33 CF patients. No bacteremia was identified. Isolates were available in 18 patients and all were genetically distinct. Two thirds of these isolates were susceptible to usual anti-pseudomonal antibiotics. After acquisition, only 40% of CF patients were chronically infected (> or =2 positive cultures separated by at least 6 months). Chronic infection was associated with resistance to > or =2 antibiotic groups on initial culture and failure of eradication after antibiotic therapy. The impact of acquisition of B. gladioli infection in chronic infection was variable. Three CF patients with chronic infection underwent lung transplantation. One post-transplant patient developed a B. gladioli mediastinal abscess, which was treated successfully. CONCLUSIONS: The majority of patients' culture positive for B. gladioli at our center have CF. B. gladioli infection is often transient and is compatible with satisfactory post-lung transplantation outcomes. PMID- 17137847 TI - GABAergic processes mediate thermal prolongation of the laryngeal reflex apnea in decerebrate piglets. AB - We tested the hypotheses that elevated body temperature would prolong reflex apnea following electrical stimulation of the superior laryngeal nerve (SLN) in decerebrate neonatal piglets and that thermal prolongation of reflex apnea after stimulation of the SLN depended on GABAergic mechanisms. These studies were conducted in 13 decerebrate piglets (age 3-15 days). The SLN was stimulated at approximately 1.5 times the threshold stimulus level for 10 s starting at the beginning of inspiration. We measured the duration of the apnea and respiratory disruption that followed SLN stimulation. Elevating body temperature prolonged the duration of the apnea and respiratory disruption that followed SLN stimulation, and treatment with antagonists of gama-aminobutyric acid A-type (GABAA) receptors reversed the thermal prolongation of reflex apnea and the period of respiratory disruption even though body temperature remained elevated. We conclude that elevated body temperature enhances or amplifies GABAergic mechanisms that prolong the respiratory inhibition following electrical stimulation of the SLN. PMID- 17137848 TI - Suicide by tapestry needle. AB - A case of completed suicide committed by a 46-year-old male by inserting a tapestry needle in his heart is described. Scene investigation findings and autopsy examination revealed signs of self-harm efforts as well as punctures at both forearms and the right and left sides of the neck. The deceased caused a penetrating cardiac injury which resulted in cardiac tamponade. PMID- 17137850 TI - Blue rubber bleb nevus syndrome. PMID- 17137849 TI - Genetic variation at the low-density lipoprotein receptor-related protein 5 (LRP5) locus modulates Wnt signaling and the relationship of physical activity with bone mineral density in men. AB - Polymorphisms in the LRP5 gene have been associated with bone mineral density (BMD) in men and/or women. However, the functional basis for this association remains obscure. We hypothesized that LRP5 alleles could modulate Wnt signaling and the relationship between physical activity and BMD. This genetic association study was performed in the population-based Framingham Study Offspring Cohort, and included a subset of 1797 unrelated individuals who provided blood samples for DNA and who had BMD measurements of the hip and spine. Ten single-nucleotide polymorphisms (SNPs) spanning the LRP5 gene were genotyped and used for association and interaction analyses with BMD by regression methods. LRP5 haplotypes were transiently co-expressed with Wnt3a, MesD and Dkk1 in HEK293 cells and their activity evaluated by the TCF-Lef reporter assay. Six out of ten SNPs in LRP5 were associated with one or more of the femur or spine BMDs in men or women after adjustment for covariates, and these associations differed between genders. In men< or =age 60 years, 3 SNPs were significantly associated with BMD: rs2306862 on Exon 10 with femoral neck BMD (p=0.01) and Ward's BMD (p=0.01); rs4988321/p. V667M with Ward's BMD (p=0.02); and intronic rs901825 with trochanter BMD (p=0.03). In women, 3 SNPs in intron 2 were significantly associated with BMD: rs4988330 for trochanter (p=0.01) and spine BMD (p=0.003); rs312778 with femoral neck BMD (p=0.05); and rs4988331 with spine BMD (p=0.04). For each additional rare allele, BMD changed by 3-5% in males and 2-4% in females. Moreover, there was a significant interaction between physical activity and rs2306862 in exon 10 (p for interaction=0.02) and rs3736228/p. A1330V in exon 18 (p for interaction=0.05) on spine BMD in men. In both cases, the TT genotype was associated with lower BMD in men with higher physical activity scores, conversely with higher BMD in men with lower physical activity scores. In vitro, TCF-Lef activity in presence of Wnt3a was significantly reduced in cells expressing LRP5 haplotypes carrying the T allele of exon 10 and 18 compared to the wild-type allele, whereas co-expression of Dkk1 completely inhibited Wnt3a response through all LRP5 haplotypes. In summary, genetic variation in exons 10 and 18 of the LRP5 gene modulates Wnt signaling and the relationship between physical activity and BMD in men. These observations suggest that Wnt-LRP5 may play a role in the adaptation of bone to mechanical load in humans, and may explain some gender-related differences in bone mass. PMID- 17137851 TI - Cholecystoduodenal fistula. PMID- 17137852 TI - A subepithelial mass determined by EUS to be a splenic artery aneurysm. PMID- 17137853 TI - Should there be light in the esophageal tunnel? An appraisal of optical coherence tomography in Barrett's esophagus. PMID- 17137854 TI - Chronic mesenteric ischemia and visible light spectroscopy: can a dark path be illuminated? PMID- 17137855 TI - Pneumatosis intestinalis in the differential diagnosis of arteriovenous malformations. PMID- 17137856 TI - A case of ileal heterotopic pancreas with repeated melena. PMID- 17137857 TI - Diagnosis of chronic mesenteric ischemia by visible light spectroscopy during endoscopy. AB - BACKGROUND: Chronic mesenteric ischemia can be difficult to diagnose by means of currently available clinical techniques. We developed a novel endoscopic device for objective measurement of GI mucosal ischemia. OBJECTIVE: Our purpose was to evaluate the performance of the device in patients with chronic mesenteric ischemia. DESIGN: A fiberoptic catheter-based visible light spectroscopy oximeter (T-Stat 303 Microvascular Oximeter, Spectros, Portola Valley, Calif) was used to evaluate 30 healthy control subjects and 3 patients with chronic mesenteric ischemia before and after successful percutaneous stenting. SETTING: Veterans Affairs Palo Alto Health Care System hospital. RESULTS: Normal mucosal (capillary) hemoglobin oxygen saturation was 60% to 73% in the duodenum and jejunum. In the 3 patients with chronic mesenteric ischemia, ischemic areas in the duodenum or proximal jejunum were found with mucosal saturations of 16% to 30%. After successful angioplasty and stent placement of the celiac, superior mesenteric, or inferior mesenteric arteries, the mucosal saturation in these areas increased to 51% to 60%. CONCLUSIONS: This preliminary study suggests that chronic mesenteric ischemia is detectable during endoscopy by use of visible light spectroscopy and that successful endovascular treatment results in near normalization of mucosal oxygen saturation. PMID- 17137858 TI - Identifying intestinal metaplasia at the squamocolumnar junction by using optical coherence tomography. AB - BACKGROUND: Optical coherence tomography (OCT) is an optical imaging method that produces high-resolution cross-sectional images of the esophagus. The accuracy of OCT for differentiating tissue types at the squamocolumnar junction (SCJ) has not been established. OBJECTIVE: The purpose of this study was to identify and validate OCT image criteria for distinguishing metaplastic from nonmetaplastic tissue at the SCJ. DESIGN: A total of 196 biopsy-correlated OCT images of the SCJ were acquired from 113 patients undergoing upper endoscopy. A pathologist blinded to the OCT results reviewed each pathology specimen and determined the presence of the following histopathology: gastric cardia, squamous mucosa, pancreatic metaplasia, and intestinal metaplasia. An algorithm for diagnosing specialized intestinal metaplasia (SIM) was created by reviewing a training set of 40 biopsy correlated OCT images. Two blinded investigators prospectively tested the algorithm on a validation set of 123 images. RESULTS: OCT images of squamous mucosa were characterized by a layered appearance without epithelial glands; gastric cardia, by vertical pit and gland structure, a well-defined epithelial surface reflectivity, and relatively poor image penetration; and SIM by an irregular architecture and good image penetration. The OCT criteria were 85% sensitive and 95% specific for SIM when applied retrospectively to the training set. When applied to the validation set, the algorithm was 81% sensitive for both OCT readers and 66% and 57% specific for diagnosing SIM. The interobserver agreement was good (kappa = 0.53). CONCLUSIONS: OCT imaging can identify SIM at the SCJ with an accuracy similar to that of endoscopy. PMID- 17137859 TI - Endoscopic argon plasma coagulation of Marlex mesh erosion after vertical-banded gastroplasty. AB - BACKGROUND: Marlex mesh erosions may occur as late complications after vertical banded gastroplasty. Experience with the endoscopic treatment is limited. OBJECTIVE: To describe the use of argon plasma coagulation in the endoscopic treatment of eroded Marlex mesh. DESIGN: Case report. SETTINGS: Endoscopy Unit, University of Iowa Hospitals and Clinics, Iowa City, Iowa, USA. PATIENTS: We describe the endoscopic treatment of eroded Marlex mesh in 2 patients who presented with symptoms of gastric-outlet obstruction. INTERVENTIONS: In both cases, argon plasma coagulation was used to break down the eroded Marlex mesh. Fragments were subsequently removed with forceps and electrocautery snares. We did not encounter any complications with this method. RESULTS: The endoscopic treatment resulted in lasting symptomatic improvement in both patients. LIMITATIONS: Our experience is limited to 2 cases. CONCLUSIONS: Argon plasma coagulation appears to be a promising option for the endoscopic treatment of eroded Marlex mesh. It allows the fragmentation of large mesh portions and enables subsequent removal with a snare and a forceps. This method can result in symptomatic improvement and may obviate the need for surgery. Further data are necessary to evaluate the safety and the efficacy of this approach. PMID- 17137860 TI - Successful treatment of gallstone ileus (Bouveret's syndrome) by using extracorporal shock wave lithotripsy and argon plasma coagulation. PMID- 17137861 TI - Candida infection presenting as multiple ulcerated masses. PMID- 17137862 TI - Small-bowel perforation after colonoscopy. PMID- 17137863 TI - Antibiotics and asthma treatment. PMID- 17137864 TI - Acorns and inhalers: the Asthma Clinical Research Network. PMID- 17137865 TI - Multiple chemical sensitivities: A systematic review of provocation studies. AB - A systematic review of provocation studies of persons reporting multiple chemical sensitivities (MCS) was conducted from databases searched from inception to May 2006. Thirty-seven studies were identified, testing 784 persons reporting MCS, 547 control subjects, and 180 individuals of whom a subset were chemically sensitive. Blinding was inadequate in most studies. In 21 studies odors of chemicals were probably apparent; 19 of these reported positive responses to provocations among chemically sensitive individuals, and 1 study demonstrated that negative expectations were significantly associated with increased symptom reporting after provocations. Seven studies used chemicals at or below odor thresholds, and 6 failed to show consistent responses among sensitive individuals after active provocation. Six studies used forced-choice discrimination and demonstrated that chemically sensitive individuals were not better at detecting odor thresholds than nonsensitive participants. Three studies tested individuals by using nose clips/face masks and confirmed response, possibly mediated through eye exposure. Three studies used olfactory masking agents to conceal stimuli, and none of these found associations between provocations and response. We conclude that persons with MCS do react to chemical challenges; however, these responses occur when they can discern differences between active and sham substances, suggesting that the mechanism of action is not specific to the chemical itself and might be related to expectations and prior beliefs. PMID- 17137866 TI - First case of homozygous C1 inhibitor deficiency. AB - BACKGROUND: C1 Inhibitor (C1-Inh) deficiency causes angioedema and can be hereditary (HAE), caused by mutations in the C1-Inh gene (C1NH), or acquired (AAE). Patients with HAE show a complement profile different from that of patients with AAE with normal levels of C1 (C1q, C1r, and C1s). OBJECTIVE: We sought to characterize the complement profile of a patient with HAE and a mutation in homozygosis in the C1NH gene (c.1576T>G, Ile462Ser) and study his family. METHODS: Biochemical diagnosis of HAE was confirmed by analyzing the C1NH gene. Further studies on the levels and activation states of the C1q, C1r, C1s, and C1-Inh components of the classical pathway of complement activation were also performed. RESULTS: Another 7 members of the family were given diagnoses of HAE: 1 was homozygous and 6 were heterozygous for the C1NH mutation c.1576T>G. The homozygous patients showed undetectable C1q levels, reduced C1s levels, the circulating active form of C1r, and a C1-Inh mostly in its cleaved inactive form in plasma. CONCLUSION: This is the first report of patients homozygous for a mutation affecting the coding region of C1NH. These patients showed a unique activation and consumption profile of the classical complement activation pathway different from that commonly observed in patients with HAE but similar to that of patients with AAE. CLINICAL IMPLICATIONS: The most common HAE treatment is attenuated androgens, which increase the C1NH gene transcription levels. Because the homozygous patients lack a wild-type allele, long-term prophylactic treatment with attenuated androgens might not be advisable. PMID- 17137867 TI - Expression of CD103 identifies human regulatory T-cell subsets. AB - BACKGROUND: Analysis of naturally occurring T regulatory CD4+ (nTreg) cells in human diseases is hampered by the lack of specific surface marker. Indeed, the CD25 antigen, which is typically used to identify nTreg cells, is also expressed on activated effector T cells. OBJECTIVE: We sought to examine whether CD4+ T cells bearing CD103 are suppressor cells, regardless of CD25 coexpression. METHODS: We first compared freshly isolated tonsillar CD103+ CD25- cells with their CD103- CD25high counterparts for their capacity to suppress T-cell response and their expression of FoxP3 mRNA. Next CD103 was induced on neonatal or adult CD4+ T cells stimulated with allogeneic dendritic cells, and the CD103+ and CD103 fractions were compared as above. RESULTS: Tonsillar CD4+ CD103+ CD25- T cells displayed comparable suppressive activity and contained similar amounts of FoxP3 mRNA as their CD103- CD25high counterparts. In vitro-generated alloantigen-primed CD103+ cells coexpressed CD25, suppressed T-cell activation, and contained more FoxP3 mRNA than the CD103- CD25+ cells isolated from the same cultures. Finally, neonatal alloreactive cells contained more CD103+ Treg cells than their adult counterparts and, unlike the latter, became hyporesponsive to the priming alloantigens. CONCLUSIONS: The examination of CD103 and CD25 coexpression allows identification of 3 subsets of human CD4+ nTreg cells, and the detection of CD103 on CD4+ T cells identifies nTreg cells, regardless of CD25 coexpression. CLINICAL IMPLICATIONS: The greater induction of CD103+ suppressor cells by cord blood should be related to its successful clinical use as an alternative to adult bone marrow transplantation. PMID- 17137868 TI - Persistent central memory phenotype of circulating Fel d 1 peptide/DRB1*0101 tetramer-binding CD4+ T cells. AB - BACKGROUND: Although substantial evidence suggests that T cells are important in the pathogenesis of atopic dermatitis (AD), little is known of the differentiation status of CD4+ T cells specific for common environmental allergens. OBJECTIVE: To determine the frequency, differentiation phenotype, and function of circulating allergen-specific CD4+ T cells in adult individuals with severe persistent AD and controls. METHODS: Using tetrameric complexes of an HLA DRB1*0101 restricted epitope from Fel d 1, the major IgE-reactive component of cat dander, we studied ex vivo and cultured T-cell frequency and phenotype in individuals with AD and healthy controls. Cytokine secretion was measured by ex vivo and cultured IFN-gamma, IL-4, and IL-10 enzyme linked immuno-spot analysis. RESULTS: Ex vivo Fel d 1-specific DRB1*0101-restricted CD4+ T cells express high levels of CCR7, CD62L, CD27, and CD28 and proportionately low levels of tissue specific homing receptors and TH1 and TH2 cytokine production, placing the cells largely within the central memory subgroup. CONCLUSION: Circulating Fel d 1 specific DRB1*0101-restricted CD4+ T cells maintain central memory capacity, consistent with a potential to contribute to persisting clinical atopic disease. CLINICAL IMPLICATIONS: Persisting central memory characteristics of allergen specific CD4+ T cells in individuals with AD may contribute to chronic disease. PMID- 17137869 TI - Parvalbumin is not responsible for cross-reactivity between tuna and marlin: A case report. PMID- 17137870 TI - Low clinical penetrance of mannose-binding lectin-associated serine protease 2 deficiency. PMID- 17137881 TI - Global child health: an essential component of residency training. PMID- 17137882 TI - Influenza vaccine for young children: two doses are better than one. PMID- 17137883 TI - Cardiovascular risk at the extremes of body composition. PMID- 17137884 TI - Midodrine for the treatment of vasovagal syncope (simple faint). PMID- 17137885 TI - Marijuana use, adolescent pregnancy, and alteration in newborn behavior: how complex can it get? PMID- 17137886 TI - Pharmacologic treatment of hypertension in children and adolescents. PMID- 17137887 TI - Influenza vaccine effectiveness in healthy 6- to 21-month-old children during the 2003-2004 season. AB - OBJECTIVE: To assess the clinical effectiveness of influenza vaccine in preventing influenza-like illness (ILI) office visits. STUDY DESIGN: We analyzed billing and immunization registry data for healthy 6- to 21-month-olds from 5 Denver, Colorado pediatric practices (n = 5193). ILI and pneumonia/influenza (a subset of ILI) were defined from International Classification of Diseases, Ninth Revision, Clinical Modification codes for office visits occurring during peak influenza season. Partially vaccinated (PV) and fully vaccinated (FV) patients were defined as having 1 shot and 2 shots, respectively, more than 14 days before the first ILI visit. The likelihood of an ILI visit was determined using a Cox proportional hazards model accounting for patient characteristics, practice site, and immunization status. RESULTS: A total of 28% of the patients had an ILI office visit, and 5% had a pneumonia/influenza visit. Hazard ratios (HRs) for FV versus UV were 0.31 (95% confidence interval [CI] = 0.3 to 0.4) for ILI and 0.13 (95% CI = 0.1 to 0.2) for pneumonia/influenza, corresponding to a vaccine effectiveness (1 - HR x 100) of 69% for ILI and 87% for pneumonia/influenza. The corresponding HRs for PV versus UV were 1.0 (95% CI = 0.9 to 1.2) and 1.1 (95% CI = 0.8 to 1.5). CONCLUSIONS: Although 2 doses of vaccine were 69% effective against ILI office visits and 87% effective against pneumonia/influenza office visits, 1 dose did not prevent office visits during the 2003-2004 influenza season. PMID- 17137889 TI - Uncoupling of cardiovascular risk markers in adolescent girls with anorexia nervosa. AB - OBJECTIVE: Cardiovascular (CV) risk begins in childhood, and low body weight should result in a favorable risk profile. However, adolescents with anorexia nervosa (AN) have alterations in many hormonal factors that mediate CV risk. We hypothesized that in AN, growth hormone (GH) resistance and hypercortisolemia would increase CV risk through effects on pro-inflammatory cytokines and lipid status despite low weight. STUDY DESIGN: We examined CV risk markers (high sensitivity C-reactive protein [hsCRP], interleukin-6 [IL-6], apolipoprotein-B [Apo-B], and lipid profile) in 23 subjects with AN and in 20 control subjects 12 to 18 years of age, in whom GH, cortisol, leptin, and triiodothyronine (T3) had been previously determined. RESULTS: Subjects with AN had higher Apo-B (P < .0001), IL-6 (P = .03), Apo-B/high-density lipoprotien (HDL) (P = .01), and Apo B/low-density lipoprotein (LDL) (P < .0001) and lower hsCRP (P = .01) than controls. Triglycerides were lower and HDL higher in subjects with AN. IGF-I predicted hsCRP in controls but not in AN. Log hsCRP correlated positively with GH and inversely with leptin. On regression modeling, the most significant predictor of log hsCRP was leptin; T3 predicted log IL-6, log Apo-B, log Apo B/HDL, and Apo-B/LDL; and cortisol independently predicted log Apo-B. IL-6 decreased with weight gain. CONCLUSION: CV risk markers are uncoupled in AN, with increased Apo-B and IL-6 and decreased hsCRP, related to hormonal alterations. IL 6 normalizes with weight gain. PMID- 17137890 TI - Correlates of violent behavior among adolescents presenting to an urban emergency department. AB - OBJECTIVE: To examine rates and correlates of self-reported perpetrated violence among teens presenting to an urban emergency department. STUDY DESIGN: Cross sectional study of a consecutive sample of youth (12-17 years) presenting to an urban emergency department. The youth were surveyed about demographics, community violence exposure, parental monitoring, substance use, weapon carriage, group fighting, and aggression (past 12 months). Bivariate and multivariate analyses were performed to identify correlates of the severity of the violence (none, moderate, or severe) perpetrated in the past year. RESULTS: A total of 115 youth were surveyed (males, 47%; response rate, 85%). The sample group was 51% African American. A total of 77% of the youth reported that they perpetrated violence in the past year, with 37% reporting severe violence (use of a weapon, group fighting, causing an injury requiring medical care). All patients who self reported perpetrating severe violence in the past year were discharged home from the emergency department. The diagnosis made in the emergency department (illness versus injury) was not associated with the violence perpetrated in the past year. Sex was not a significant predictor of severe perpetrated violence. CONCLUSIONS: Youth presenting to urban emergency departments report high rates of perpetrating violence in the past year that, on the basis of other studies, is likely to continue. These results suggest that future emergency department-based violence prevention efforts should consider universal screening of adolescents. PMID- 17137891 TI - The efficacy of midodrine hydrochloride in the treatment of children with vasovagal syncope. AB - OBJECTIVE: To determine whether midodrine hydrochloride therapy can prevent vasovagal syncope (VVS) in pediatric patients. STUDY DESIGN: Children with recurrent syncope (n = 26) were randomly assigned into 2 groups. Group I comprised children given midodrine hydrochloride as first-line therapy in addition to conventional therapy, and group II comprised patients receiving conventional therapy only. Repeat head-up tilt (HUT) testing and follow-up of least 6 months were conducted to evaluate the therapeutic effectiveness and side effects of midodrine in treating VVS in children. RESULTS: The HUT-based effective rate was significantly higher in group I than in group II (75% vs 20%; P < .05). During the follow-up period, the recurrence of syncope was significantly lower in group I than in group II (P < .05). CONCLUSIONS: Midodrine hydrochlorate is effective in treating VVS in children, especially in preventing recurrent episodes. Few side effects were observed in the present study. PMID- 17137892 TI - Exposure to marijuana during pregnancy alters neurobehavior in the early neonatal period. AB - OBJECTIVE: To assess the neurobehavior of full-term neonates of adolescent mothers exposed to marijuana during pregnancy. STUDY DESIGN: This prospective cross-sectional study included full-term infants within 24 to 72 hours of life born to adolescent mothers at a single center in Brazil. Data on sociodemographic and obstetrical and neonatal characteristics were collected. The mothers underwent the Composite International Diagnostic Interview, and the infants were assessed with the Neonatal Intensive Care Unit Network Neurobehavioral Scale (NNNS). Maternal hair and neonatal meconium were analyzed. Neonates exposed in utero to tobacco, alcohol, cocaine, and/or any other drugs except marijuana were excluded. RESULTS: Of 3685 infants born in the study hospital, 928 (25%) were born to adolescent mothers. Of these, 561 infants met the inclusion criteria and were studied. Marijuana exposure was detected in 26 infants (4.6%). Infants exposed (E) or not exposed (NE) to marijuana differed in the following NNNS variables: arousal (E, 4.05 +/- 1.19 vs NE, 3.68 +/- 0.70), regulation (E, 5.75 +/- 0.62 vs NE, 6.04 +/- 0.72), and excitability (E, 3.27 +/- 1.40 vs NE, 2.40 +/ 1.57). After controlling for confounding variables, the effect of marijuana exposure on these scores remained significant. CONCLUSIONS: Marijuana exposure during pregnancy alters the neurobehavioral performance of term newborn infants of adolescent mothers. PMID- 17137893 TI - Comparison of chronic illness among children receiving mechanical ventilation in a cohort of children's hospitals in 1991 and 2001. AB - OBJECTIVE: To compare the prevalence of chronic illness and characteristics of children who underwent mechanical ventilation in a cohort of patients at large children's hospitals between 1991 and 2001. STUDY DESIGN: This was a retrospective, cross-sectional study using the National Association of Children's Hospitals and Related Institutions (NACHRI) case mix database to identify children who underwent mechanical ventilation and compare the prevalence of chronic illness and incidence of mechanical ventilation. RESULTS: The proportion of children who underwent mechanical ventilation who had at least 1 chronic condition increased from 72% in 1991 to 75% in 2001. The incidence of mechanical ventilation in hospitalized children almost doubled during this decade, from 77 per 1000 hospitalizations in 1991 to 124 per 1000 in 2001. The rate of mechanical ventilation increased with increasing numbers of chronic conditions. The mortality of children who underwent mechanical ventilation decreased from 14% in 1991 to 11% in 2001. CONCLUSIONS: The increase in mechanical ventilation in hospitalized children is due to both an increased incidence of chronic illness and higher use within diagnostic categories. Unlike utilization of some services, the use of mechanical ventilation in hospitalized children may be a marker of increased severity of illness and need, because it is plausible that mechanical ventilation use is not primarily supply-sensitive. PMID- 17137894 TI - Lung bioavailability of hydrofluoroalkane fluticasone in young children when delivered by an antistatic chamber/mask. AB - OBJECTIVE: To determine whether an antistatic valved holding chamber/mask improves lung bioavailability of hydrofluoroalkane (HFA) fluticasone in young children. STUDY DESIGN: Twelve patients, age 1 to 6 years, with well-controlled asthma were treated with an HFA fluticasone metered-dose inhaler (Flovent HFA) twice daily (440 microg/day). The drug was delivered by tidal breathing through conventional (AeroChamber Plus) and antistatic (AeroChamber MAX) valved holding chambers (VHCs) with masks in a randomized, crossover manner, each for 3 to 7 days. When adherence was 100% at home, blood was collected for measurement of steady-state fluticasone plasma concentration (FPC) 1 hour after the last dose was administered in the clinic. FPC indicates systemic exposure directly and airway delivery indirectly. It was measured by liquid chromatography-mass spectrometry. Data were analyzed by regression analysis. RESULTS: The mean +/- SD FPC was 107 +/- 30 pg/mL after conventional VHC and 186 +/- 134 pg/mL after the antistatic VHC (P = .03). In 5 patients (40%), the antistatic VHC increased FPC by >/= 100%, to potentially excessive levels in 4 of them; it had little effect in 7 patients. CONCLUSIONS: HFA fluticasone was delivered to the airways by both devices even though the patients could not inhale deeply and breath hold. The antistatic VHC variably increased lung bioavailability. To reduce systemic exposure, the dose should be weaned to the minimum required to maintain asthma control. PMID- 17137895 TI - Leptin levels decline steadily during prolonged fasting in lean children. AB - OBJECTIVE: To evaluate the effects of fasting on serum leptin levels in lean children. STUDY DESIGN: Seventeen children, age 7.7 +/- 4.3 years with mean body mass index (BMI) of 16.7 +/- 2.7 kg/m(2), underwent standard diagnostic fasts for suspected hypoglycemia. Blood was sampled at 6-hour intervals for glucose, insulin, C-peptide, leptin, free and total insulin-like growth factor-1, insulin like growth factor-binding protein-1, growth hormone, cortisol, ketones, and free fatty acids. RESULTS: Subjects fasted 15 to 40 hours, and initial leptin levels were related to BMI and age. Leptin declined by 0.5 ng/mL per each fasting hour (P = .008), using a longitudinal mixed effects model. Leptin dropped significantly from an initial mean +/- SEM during the first 6 hours of 15.9 +/- 5.5 ng/mL to 3.5 +/- 0.9 ng/mL at the end of fasting. Mixed longitudinal effects models demonstrated that leptin was significantly related to insulin over time (P < .0001) as well as C-peptide (P < .0001). Significant relations were also seen with total insulin-like growth factor-1 over time, beta-hydroxybutyrate and insulin, and insulin-like growth factor-binding protein-1 and insulin. CONCLUSIONS: After 6 hours, leptin levels steadily decline during prolonged fasting in lean children. The decline probably is related to the suppression of insulin secretion. Although baseline leptin levels were related to BMI and age, in the final fasting sample, leptin levels showed minimal variation in this pediatric cohort encompassing a wide age range. PMID- 17137896 TI - Persistence of obstructive sleep apnea syndrome in children after adenotonsillectomy. AB - OBJECTIVE: To investigate the relative contribution of various risk factors to the surgical outcome of adenotonsillectomy for obstructive sleep apnea syndrome in children. STUDY DESIGN: Children (n = 110; mean age, 6.4 +/- 3.9 years) underwent two polysomnographic evaluations before and after adenotonsillectomy. In addition, 22 control children were studied. History for allergy and family history of sleep-disordered breathing was taken before each polysomnographic evaluation. RESULTS: Significant changes in sleep stage percentages and sleep fragmentation were found in the postsurgery study compared with the presurgery study; 25% of the children had apnea/hypopnea index (AHI) 1 and <5, and 29% had AHI >/=5 in the postsurgery study. The frequency of subjects with AHI 0.9 U/kg/d), and suboptimal metabolic control (A1c 7.5%-11%), with the primary outcome of change in A1c. Secondary outcomes include change in insulin dose, body mass index (BMI), lipids, and waist and hip circumference. RESULTS: Metabolic control (A1c) was improved at 6 months in all subjects (P = .02). There was no significant difference between the pioglitazone and placebo treatment groups at 6 months in either change in A1c (-0.4% +/- 0.9% and -0.5% +/- 1.2%, respectively) or insulin dose. BMI SDS increased by 0.3 +/- 0.3 (kg/m(2)) in the pioglitazone group and remained unchanged in the placebo group (P = .01). There was no significant difference in change in any lipid parameters between the pioglitazone and placebo groups at 6 months. CONCLUSIONS: Adjunctive pioglitazone therapy was not effective in improving glycemic control in adolescents with T1D. Pioglitazone was associated with increased BMI. PMID- 17137906 TI - Familial association of pleuropulmonary blastoma with cystic nephroma and other renal tumors: a report from the International Pleuropulmonary Blastoma Registry. AB - OBJECTIVE: To characterize the association of pleuropulmonary blastoma (PPB) with cystic nephroma (CN) and other renal tumors. STUDY DESIGN: Complete clinicopathologic review of cases from the International PPB Registry and literature. RESULTS: We identified 18 patients with PPB associated with 20 renal tumors (15 CN), either in themselves or family members. All patients with PPB were <5 years of age. All but one of the renal diagnoses were made before 4 years of age. Eleven children had both PPB and renal tumor, one of whom also had a sibling with CN. Six children with PPB alone had one or more family members with CN. The mother of one child with PPB had Wilms' tumor. Pulmonary disease was bilateral in four patients. Renal disease was bilateral in three patients. Two children with PPB and bilateral renal cystic tumors also had intussusceptions because of small bowel juvenile polyps. In six families, dysplasia/neoplasia affected organs other than lung and kidney. CONCLUSIONS: CN or related tumors were found in 9.2% of 152 Registry-reviewed PPB cases. The occurrence of rare pulmonary and renal tumors together in patients and/or family members, the early age of onset, and the multiplicity of tumors is compatible with a constitutional genetic predisposition. PMID- 17137907 TI - Minimal risk in pediatric research. PMID- 17137908 TI - Involving children with cancer in decision-making about research participation. PMID- 17137909 TI - Pediatric workforce: a look at pediatric rheumatology data from the American Board of Pediatrics. PMID- 17137910 TI - Increased farnesylation of p21-Ras and neonatal macrosomia in women with gestational diabetes. AB - Six of 22 mothers with gestational diabetes mellitus had infants with macrosomia, cord blood hyperinsulinemia, and increased amounts of a key mitogenic intermediate, farnesylated p21-Ras. The ability of fetal hyperinsulinemia to increase the availability of farnesylated p21-Ras may represent one mechanism of the growth-promoting action of insulin during fetal development. PMID- 17137911 TI - Normal minipuberty of infancy in boys with Prader-Willi syndrome. AB - In 4 boys with Prader-Willi syndrome, 3 of whom had cryptorchidism, penile length and plasma testosterone levels at 5 to 10 weeks of life were normal. We suggest that cryptorchidism reflects decreased abdominal pressure rather than hypogonadism, which develops later from progressive degeneration of gonadotropin releasing hormone neurons. PMID- 17137912 TI - Presentation and clinical progression of pseudohypoparathyroidism with multi hormone resistance and Albright hereditary osteodystrophy: a case series. AB - Anthropometric and biochemical features were retrospectively evaluated in 12 patients with pseudohypoparathyroidism, Albright hereditary osteodystrophy, and multi-hormone resistance. Hypothyroidism and subcutaneous calcifications were presenting features in younger children. Temporal trends in stimulatory hormone resistance included early thyroid-stimulating hormone elevation and progression from parathyroid hormone elevation to hyperphosphatemia and hypocalcemia. PMID- 17137913 TI - Childhood steroid-responsive painful opthalmoplegia: clues to opthalmoplegic migraine. PMID- 17137914 TI - Requirement for screening for vesicoureteric reflux in children with multicystic dysplastic kidney: some considerations based on our experience and literature review. PMID- 17137916 TI - Percutaneous gastrostomy feeding in patients with cystic fibrosis. PMID- 17137918 TI - Diagnosing urinary tract infections in young children. PMID- 17137921 TI - Introduction on the use of the Drosophila embryonic/larval neuromuscular junction as a model system to study synapse development and function, and a brief summary of pathfinding and target recognition. PMID- 17137922 TI - Development and structure of motoneurons. PMID- 17137923 TI - The development of the Drosophila larval body wall muscles. PMID- 17137924 TI - Organization of the efferent system and structure of neuromuscular junctions in Drosophila. PMID- 17137925 TI - Development of motoneuron electrical properties and motor output. PMID- 17137926 TI - Transmitter release at the neuromuscular junction. PMID- 17137927 TI - Vesicle trafficking and recycling at the neuromuscular junction: two pathways for endocytosis. PMID- 17137928 TI - Glutamate receptors at the Drosophila neuromuscular junction. PMID- 17137929 TI - Scaffolding proteins at the Drosophila neuromuscular junction. PMID- 17137930 TI - Synaptic cytoskeleton at the neuromuscular junction. PMID- 17137932 TI - Retrograde signaling that regulates synaptic development and function at the Drosophila neuromuscular junction. PMID- 17137931 TI - Plasticity and second messengers during synapse development. PMID- 17137933 TI - Activity-dependent regulation of transcription during development of synapses. PMID- 17137934 TI - Experience-dependent potentiation of larval neuromuscular synapses. PMID- 17137935 TI - Selected methods for the anatomical study of Drosophila embryonic and larval neuromuscular junctions. PMID- 17137937 TI - Prevalence of myocardial ischemia during mechanical ventilation and weaning and its effects on weaning success. AB - BACKGROUND: Myocardial ischemia may be considered both a consequence of weaning from ventilation and a potential cause of weaning failure. A limited number of investigations have evaluated myocardial ischemia during mechanical ventilation and weaning and its effect on weaning success. The purpose of this pilot investigation was to determine the prevalence of myocardial ischemia in a diverse group of medical intensive care unit (MICU) patients during baseline mechanical ventilation, during weaning using continuous positive airway pressure, and up to 24 hours after extubation and to evaluate the relationship between ischemia and weaning failure. METHODS: This study was a prospective, repeated-measure, descriptive investigation that studied 43 ventilated MICU patients. Slightly more than half of participants were male (53%), and participants had a mean age of 51.1 +/- 14.6 years and were ventilated an average of 11.7 +/- 11.3 days. Myocardial ischemia was evaluated by examination of plasma cardiac troponin I, creatine phosphokinase-myocardial band (CK-MB), and ST-segment changes on electrocardiogram. Continuous electrocardiographic data were obtained by a calibrated, frequency-modulated, continuous 3-channel electrocardiographic recorder using leads I, II, and V2. RESULTS: Seventy percent of these participants (n = 30) exhibited ST-segment deviation at some point during data collection (baseline mechanical ventilation, during weaning, after extubation). Twenty-four participants exhibited ST-segment deviation at baseline, 7 during weaning, and 8 after extubation. Nine participants exhibited ST-segment deviation at >1 data collection time point. None had ST-segment deviation at all 3 time points. Cardiac enzyme concentrations were highly variable; five participants demonstrated clinically important increases in either CK-MB and/or troponin I. Thirty-five percent of participants required >1 weaning trial. Participants who exhibited ST-segment deviation during baseline ventilation were 60% more likely to fail their initial weaning trial. There were no significant differences in CK MB or troponin I levels between those participants who were successfully weaned with 1 trial and those who failed to wean during that first trial. CONCLUSIONS: Silent myocardial ischemia was a common occurrence in this diverse group of MICU patients, although only 21% had previously diagnosed coronary disease. Clinicians must be aware of the potential for silent ischemia, monitor and evaluate their patients for such, and intervene to promote optimal cardiovascular function, particularly during the stress of ventilator weaning. PMID- 17137938 TI - Internet-based behavioral change and psychosocial care for patients with cardiovascular disease: a review of cardiac disease-specific applications. AB - BACKGROUND: Web-based interventions (WBIs) seem to be an efficacious method of addressing behavioral and psychosocial maladjustment among patients with chronic illness (eg, diabetes), behavioral concerns (eg, smoking cessation), and psychologic maladjustment (eg, panic disorder). Cardiology seems particularly well suited to the application of WBIs given its strong association with behavioral (eg, lifestyle) and cognitive (eg, stress and coping) factors. To date, few studies have been published that explore the use of WBIs on psychosocial and quality of life variables among populations with cardiac disease. OBJECTIVE: The purpose of this study is to review the use of WBIs in patients with cardiovascular disease to date, and to provide recommendations as to how future WBIs can be developed specifically to meet the needs of different populations with heart disease (eg, recipients of implantable cardioverter defibrillators, patients with congestive heart failure, and patients with congenital heart disease). CONCLUSIONS: We conclude that, although limitations exist, WBIs are a reasonable modus of improving patient outcomes. PMID- 17137939 TI - Exercise and heart rate recovery. AB - PURPOSE: This study examines whether heart rate recovery (HRR) improves as a result of exercise training during cardiac rehabilitation (CR). METHODS: A retrospective study was performed that included 100 patients who completed phase II CR and had entry and exit exercise stress tests. HRR was compared for the sample. Improvements in HRR were compared between gender and age groups. Correlation between age and HRR was performed. RESULTS: The total sample improved HRR (P = .020). There was no significant difference in the improvement of HRR based on gender, indicating males and females improve at similar rates (P = .833). Similarly, there was no significant difference in the improvement of HRR based on age, indicating older subjects improve similarly to younger subjects (P = .700). There was no relationship between age and HRR; therefore, as age increases there is no decrease in HRR. CONCLUSION: HRR improves in patients who complete CR. PMID- 17137940 TI - Subjective assessment of backrest elevation: magnitude of error. AB - BACKGROUND: Backrest elevation, defined as the angle of the backrest height above the horizontal position, is a common nursing intervention that is often used by subjective visual estimation in critically ill patients. OBJECTIVES: The aim of the study was to describe the magnitude of error during the subjective assessment of backrest elevation. METHODS: This prospective study was conducted in a sample of 160 subjects: 97 registered nurses, 48 undergraduate nursing students, and 15 nursing assistants. Data were collected by recording the degrees of backrest elevation identified by the subjects through an individual random presentation of the selected study angles of 20 degrees, 30 degrees, 35 degrees, 40 degrees, and 45 degrees. A measurement instrument was developed for determination of the angles. RESULTS: Of the 800 investigated angles, 14.9% were estimated accurately, 61.6% were overestimated, and 23.5% were underestimated, with an error average of 8 degrees (+/-13.5 degrees). It was determined that the larger the angle estimated, the greater the average error. A statically significant difference (P 26.0 mm, respectively). RESULTS: No formula was more accurate than the others as measured by mean absolute error. The formulas were also equally accurate when eyes were stratified by axial length. CONCLUSION: The 4 IOL power formulas provided equivalent refractive results in the entire group of eyes and in the subsets of axial lengths tested. PMID- 17137983 TI - Clinical outcomes of combined sutureless vitrectomy with triamcinolone stain to manage vitreous loss resulting from posterior capsule rupture during phacoemulsification. AB - PURPOSE: To investigate the efficacy of sutureless pars plana vitrectomy (PPV) combined with intracameral triamcinolone stain in the management of vitreous loss associated with phacoemulsification. SETTING: Department of Ophthalmology, Tri Service General Hospital, Taipei, Taiwan, Republic of China. METHODS: This retrospective review comprised the charts of 21 patients who had sutureless PPV combined with intracameral triamcinolone stain to manage vitreous loss resulting from posterior capsule rupture during phacoemulsification. The charts were analyzed for type of cataract, posterior segment pathology, methods of anesthesia, intraocular lens (IOL) placement, postoperative visual acuity, intraocular pressure, and complications. Additional outcome measurements were duration of the surgical procedures, period of postoperative corneal edema, and time to achieve stable vision. RESULTS: Excluding 2 eyes with preexisting conditions, 18 of 19 eyes (94.7%) had a final best corrected visual acuity (BCVA) of 20/40 or better and 42.1% (8/19) had a final BCVA of 20/20 or better. The mean duration of the surgery was 25.3 minutes (range 16 to 40 minutes). Corneal edema was noted in 12 eyes (57.1%) 3 days postoperatively and 3 eyes (14.3%) at 7 days. Eleven eyes (52.4%) had stable vision at 1 week, and 16 eyes (76.2%) had stable vision within 1 month postoperatively. Four eyes (19.0%) had postoperative complications that included a displaced IOL in 3 eyes (14.3%) and cystoid macular edema in 1 eye (4.8%). CONCLUSIONS: Self-sealing, sutureless PPV combined with intracameral triamcinolone stain was a safe, reliable adjunct to manage vitreous loss during phacoemulsification. The surgery led to rapid visual recovery. PMID- 17137984 TI - Posterior-assisted levitation: outcomes in the retrieval of nuclear fragments and subluxated intraocular lenses. AB - PURPOSE: To report the outcomes of posterior-assisted levitation (PAL) to retrieve nuclear fragments and subluxated intraocular lenses (IOLs). SETTING: Singapore National Eye Centre, Singapore, Singapore. METHODS: In a retrospective series, the case notes of consecutive surgeries performed by a single surgeon between 1999 and 2006 were reviewed. Posterior-assisted levitation was performed for intraoperative posterior capsule rupture with nucleus-IOL dislocation or for late subluxation of posterior chamber intraocular lenses (PC IOLs). This was accomplished by the pars plana insertion of a needle followed by levitation of the subluxated object. Ophthalmic viscosurgical device injection was not used as an adjunct for levitation. RESULTS: Fourteen patients with PAL were followed for a mean of 18.71 months (range 1 to 79 months). There were 3 cases of dropped nucleus/nuclear fragments, 1 case of intraoperative subluxated PC IOL, and 10 cases of late subluxated PC IOLs. Complications included 1 case of retinal detachment occurring 20 months after PAL and 1 case of cystoid macular edema in a patient with previous anterior uveitis. Best corrected visual acuity was 20/40 or better in all except 3 patients (1 retinal detachment, 1 corneal edema/scarring, 1 unspecified). CONCLUSION: Posterior-assisted levitation provided an effective, relatively noninvasive means of levitating nuclear fragments and PC IOLs with few complications. PMID- 17137985 TI - Normal-eye Zernike coefficients and root-mean-square wavefront errors. AB - PURPOSE: To compare aberrometry measurements from multiple sites and compute mean Zernike coefficients and root-mean-square (RMS) values for the entire data pool to serve as a reference set for normal, healthy adult eyes. SETTING: Northeastern State University, Tahlequah, Oklahoma, USA. METHODS: Data were collected from 10 laboratories that measured higher-order aberrations (HOAs) in normal, healthy adult eyes using Shack-Hartmann aberrometry (2560 eyes of 1433 subjects). Signed Zernike coefficients were scaled to pupil diameters of 6.0 mm, 5.0 mm, 4.0 mm, and 3.0 mm and corrected to a common wavelength of 550 nm. The mean signed and absolute Zernike coefficients across data sets were compared. Then, the following were computed: overall mean values for signed and absolute Zernike coefficients; polar Zernike magnitudes and RMS values for coma-like aberrations (Z(3)(+/-1) and Z(5)(+/-1) combined); spherical-like aberrations (Z(4)(0) and Z(6)(0) combined); and 3rd-, 4th-, 5th-, and 6th-order, and higher-order aberrations (orders 3 to 6). RESULTS: The different data sets showed good agreement for Zernike coefficients values across most higher-order modes, with greater variability for Z(4)(0) and Z(3)(-1). The most prominent modes and their mean absolute values (6.0-mm pupil) were, respectively, Z(3)(-1) and 0.14 microm, Z(4)(0) and 0.13 microm, and Z(3)(-3) and 0.11 microm. The mean total higher-order RMS was 0.33 microm. CONCLUSIONS: There was a general consensus for the magnitude of HOAs expected in normal adult human eyes. At least 90% of the sample had aberrations less than double the mean values reported here. These values can serve as a set of reference norms. PMID- 17137986 TI - Transient light-sensitivity syndrome after laser in situ keratomileusis with the femtosecond laser Incidence and prevention. AB - PURPOSE: To describe the incidence of transient light-sensitivity syndrome (TLSS) after laser in situ keratomileusis (LASIK) with the femtosecond laser and to identify preventive strategies. SETTING: Hospital NISA Virgen del Consuelo, Valencia, Spain. METHODS: The first 765 eyes operated on with the 15 KHz femtosecond laser were prospectively analyzed for subjective complaints and clinical findings compatible with TLSS. Intraoperative settings, postoperative treatment, and development of complications were analyzed. RESULTS: Overall, TLSS developed in 10 eyes (incidence 1.3%). However, the incidence decreased from 2.8% to 0.4% when aggressive topical steroids were used during the first 3 postoperative days. Postoperative interface inflammation and postoperative use of a low-dose topical steroid regimen were associated with a higher incidence of TLSS. CONCLUSIONS: Transient light-sensitivity syndrome is a relatively uncommon complication related to the use of the femtosecond laser. Postoperative interface inflammation may increase the probability of developing TLSS, whereas an aggressive postoperative steroid regimen seemed to provide protection against it. PMID- 17137987 TI - Comparison of residual stromal bed and flap thickness in primary and repeat laser in situ keratomileusis in myopic patients. AB - PURPOSE: To compare the change in residual stromal thickness and flap thickness between primary laser in situ keratomileusis (LASIK) and repeat LASIK in myopic patients. SETTING: Melbourne Excimer Laser Group, East Melbourne, Australia. METHODS: This retrospective nonrandomized comparative trial comprised 46 eyes of 34 patients who had repeat LASIK. The thickness of the residual stromal bed was calculated by subtracting the calculated stromal ablation from pachymetry of the stromal bed after cutting the flap in primary treatment and directly measuring during retreatment. The thickness of the LASIK flap in primary and repeat LASIK was calculated by subtracting the central pachymetry of the stromal bed after creating the flap from pachymetry before cutting and lifting the flap, respectively. The main outcome measures were comparison of the residual stromal bed and flap thickness between the primary treatment and the retreatment. RESULTS: The mean thickness of the calculated residual stromal bed after primary treatment was 329.8 microm +/- 40.8 (SD), and the mean measured residual stromal bed at retreatment was 317.3 +/- 42.8 microm. The mean difference in residual stromal bed thickness was 12.5 +/- 13.0 microm (P<.001). Sixteen eyes (34.7%) had a decrease in bed thickness between 11 microm and 20 microm. The mean flap thickness during primary LASIK and repeat LASIK was 145.2 +/- 17.1 microm and 169 +/- 18.3 microm, respectively. The mean interval between primary treatment and retreatment was 7.4 +/- 4.1 months. The mean change in flap thickness was 23.8 +/ 15.2 microm (P<.001). Fifteen eyes (32%) had an increase in flap thickness between 11 microm and 20 microm. There was a negative correlation between refractive error before primary treatment and the difference in flap thickness. No correlation was found between the difference in flap thickness and the interval between the primary treatment and the repeat treatment. CONCLUSIONS: Intraoperative pachymetry of the stromal bed during retreatment is strongly recommended as the residual stromal bed and flap thickness changes between primary and retreatment. There is a tendency for the measured stromal bed at retreatment to be thinner than the calculated stromal bed and for the flap to be thicker than previously measured. PMID- 17137988 TI - Cumulative neodymium:YAG laser rates after bag-in-the-lens and lens-in-the-bag intraocular lens implantation: comparative study. AB - PURPOSE: To study the cumulative neodymium:YAG (Nd:YAG) laser rate after bag-in the-lens implantation (Morcher 89A) and lens-in-the-bag implantation (Morcher 92S) of 2 intraocular lenses (IOLs) of the same biomaterial. SETTING: Department of Ophthalmology, University Hospital of Antwerp, Edegem, Belgium. METHODS: This study comprised 100 eyes of 87 patients who had the bag-in-the-lens IOL implantation between January 2000 and August 2004. The postoperative follow-up ranged between 17 and 72 months. One hundred eyes of 94 patients of the same age and with the same follow-up period received the lens-in-the-bag IOL. The cumulative Nd:YAG laser frequency rates in both groups were calculated, and the cumulative incidence rates were defined by Kaplan-Meier survival analysis. RESULTS: No Nd:YAG laser capsulotomy was performed in eyes having bag-in-the-lens IOL implantation. A laser capsulotomy was performed in 20 eyes having lens-in-the bag IOL implantation; the cumulative frequency in this group was 2% at 1 year and 20% at 71 months, with a plateau beginning at 42 months. The cumulative incidence rate of Nd:YAG posterior capsulotomy was approximately 2% at 1 year, increasing to approximately 28% at 42 months. CONCLUSIONS: The cumulative Nd:YAG laser rate after bag-in-the-lens implantation was zero. A zero rate has not been reported with lens-in-the-bag implantation of an IOL of the same biomaterial or of other biomaterials, as published in the literature. Thus, it can be concluded that the bag-in-the-lens implantation technique has 100% effectiveness against posterior capsule opacification. PMID- 17137989 TI - Intraocular lens power in bilateral cataract surgery: whether adjusting for error of predicted refraction in the first eye improves prediction in the second eye. AB - PURPOSE: To assess whether the retrospectively calculated intraocular lens (IOL) position value in the first eye reduces the error of predicted refraction in the second. SETTING: Prince of Wales Hospital, Sydney, Australia. METHODS: One hundred twenty-one consecutive patients who had bilateral cataract surgery with the same IOL (SI-30NB, Advanced Medical Optics) were identified. The case-derived A-constant in the first eye was calculated from the postoperative refraction. This value was used to calculate the adjusted error of predicted refraction in the second eye and compared against the unadjusted error in that eye (calculated using manufacturer's A-constant). RESULTS: Axial length (r = 0.97), corneal power (r = 0.97), and IOL power (r = 0.90) were strongly correlated between eyes with no statistically significant mean interocular difference. Although there was no significant interocular difference in the mean error of predicted refraction (SRK/T), there was only a moderate correlation between eyes (r = 0.40). Using the axial length vergence formula, the mean adjusted error of predicted refraction in the second eye (-0.66 diopter [D]) was significantly larger than the mean unadjusted error (-0.47 D) (P = .029). The standard deviation of the adjusted error of predicted refraction (SRK/T) in the second eye (0.85 D) was greater than the standard deviation of the unadjusted error (0.79). Similarly, the adjusted mean absolute error of predicted refraction (0.65 D) was greater than the unadjusted error (0.63 D). CONCLUSION: Adjusting the IOL power in the second eye by the amount of overprediction or underprediction in the first eye did not improve prediction accuracy because the error of predicted refraction varied independently between the 2 eyes of an individual. PMID- 17137990 TI - Scheimpflug measurement of intraocular lens position after piggyback implantation of foldable intraocular lenses in eyes with high hyperopia. AB - PURPOSE: To investigate the position of 3-piece foldable intraocular lenses (IOLs) after piggyback implantation for high hyperopia. SETTING: University Eye Hospital, Johann Wolfgang Goethe University, Frankfurt am Main, Germany. METHODS: Eight eyes of 5 highly hyperopic patients had phacoemulsification and implantation of 2 foldable IOLs. In 3 eyes, both IOLs were implanted in the capsular bag. In 5 eyes, 1 IOL was placed in the capsular bag and the second IOL in the ciliary sulcus. Intraocular lens optic tilt and decentration, combined thickness of both IOLs, and anterior chamber depth (ACD) were measured postoperatively over a period of 18 months using Scheimpflug photography. RESULTS: All eyes with both IOLs in the capsular bag showed interpseudophakic opacification, with a mean increase in combined IOL thickness of 0.4 mm, a decrease in ACD of 0.3 mm, and a corresponding hyperopic shift of 4.00 diopters. Eyes in which the anterior IOL was placed in the ciliary sulcus showed no changes in refraction or combined IOL thickness. In these eyes, the anterior IOL had a higher mean decentration (0.49 mm +/- 0.20 [SD] after 12 months) than the posterior IOL (0.21 +/- 0.13 mm after 12 months). CONCLUSIONS: Piggyback IOL implantation with placement of 2 foldable IOLs in the capsular bag can be followed by a hyperopic shift that may be caused in part by displacement of the IOLs. Placement of the anterior IOL in the ciliary sulcus can lead to higher decentration of this IOL. PMID- 17137991 TI - Determining whether delayed nonarteritic ischemic optic neuropathy associated with cataract extraction is a true entity. AB - PURPOSE: To evaluate cases of delayed nonarteritic anterior and posterior ischemic optic neuropathy after cataract extraction and to evaluate the need for centralized prospective reporting of nonarteritic ischemic optic neuropathy after cataract extraction. SETTING: Neuro-ophthalmology Clinics, University of Colorado Health Sciences Center and Denver Veterans Affairs Medical Center, Denver, Colorado, USA. METHODS: A retrospective review of all patients referred to the Neuro-ophthalmology Divisions, University of Colorado Health Sciences Center and Denver Veterans Affairs Medical Center, from January 2001 to October 2005 was performed. All patients with a diagnosis of nonarteritic anterior or posterior ischemic optic neuropathy were identified. Patients with ischemic optic neuropathy that occurred between 2 months and 12 months after cataract extraction were selected for evaluation. RESULTS: Six eyes with nonarteritic ischemic optic neuropathy that occurred 2 to 6 months after cataract extraction in 4 patients (2 bilateral nonarteritic ischemic optic neuropathy) and 1 eye with nonarteritic ischemic optic neuropathy that occurred after 6 months in 1 patient were identified. One patient had nonarteritic ischemic optic neuropathy in 1 eye 3 months after cataract extraction and 4 years later had nonarteritic ischemic optic neuropathy in the fellow eye 5 months after cataract extraction. Follow-up after the last cataract extraction was 1 to 3 years for all patients. CONCLUSIONS: Although an association between cataract extraction and delayed nonarteritic anterior and posterior ischemic optic neuropathy has been suggested, current data do not support a causal relationship. In addition, the window of postoperative susceptibility for delayed ischemic optic neuropathy after cataract extraction is unclear. PMID- 17137992 TI - Hyperopic shift in refraction associated with implantation of the single-piece Collamer intraocular lens. AB - PURPOSE: To describe reports of refractive hyperopic shift over time following intraocular lens (IOL) implantation, discuss possible etiologies, and suggest preventive and therapeutic treatments. SETTING: Multiple U.S. ophthalmic surgical centers; data collection at Staar Surgical, Monrovia, California, USA. METHODS: Forty cases displaying a progressive hyperopic shift in refraction after implantation of the single-piece Collamer IOL (Staar Surgical) were reported to the manufacturer out of 160 000 that have been implanted since its commercial introduction in April 2000. A retrospective data analysis of these 40 cases was performed; parameters included the mean refractive change over time, presence of capsular fibrosis, IOL displacement, and outcomes of secondary treatments. Individual case examples are presented. RESULTS: The mean refractive shift was 1.81 diopters (D) (range 0.25 to 3.75 D) with 5 cases (14%) demonstrating a hyperopic shift greater than 3.0 D. In 38 of the 40 cases, some combination of the following were reported: posterior displacement of the IOL, capsule fibrosis, and/or relief of the condition by performance of a radial or circumferential anterior neodymium:YAG capsulotomy to relieve capsule tension. The use of a small (less than 5.5 mm) capsulorhexis was also associated with cases showing the hyperopic shift. CONCLUSION: The most likely etiology was the development of anterior capsule fibrosis, sometimes exacerbated by a small capsulorhexis, which could cause the IOL to move posteriorly, resulting in a hyperopic change in refraction. Previous in vitro testing by the manufacturer ruled out a change in the refractive power of the IOL in the eye as a cause of this phenomenon. PMID- 17137993 TI - Burst-shot infrared digital photography to determine scotopic pupil diameter. AB - PURPOSE: To evaluate the usefulness of infrared digital photography for determining scotopic pupil size by comparing infrared digital photography with a Colvard pupillometer (Oasis Medical). SETTING: Department of Ophthalmology, Ajou University School of Medicine, Suwon, Korea. METHODS: Scotopic pupil size in 50 eyes of 25 healthy individuals was measured with a Colvard pupillometer and a digital camera (DSC-F828) using infrared burst shots after 5 minutes of dark adaptation. Measurements were performed by 2 independent examiners (E1 and E2). The digital photograph images were read using the ruler function of Adobe Photoshop 7.0 by 2 independent readers (R1 and R2). Agreement and repeatability were analyzed using the comparison method described by Bland and Altman. RESULTS: The mean scotopic pupil diameter measured using the Colvard pupillometer was 6.69 mm +/- 0.78 (SD) (E1) and 6.70 +/- 0.71 mm (E2). The mean scotopic pupil diameter measured from the digital photograph images was 6.67 +/- 0.75 mm (E1) and 6.66 +/ 0.78 mm (E2). The mean difference between E1 and E2 with both devices was small; however, the result with the infrared digital camera was marginally smaller than with the Colvard pupillometer. The limits of agreement were -0.01 +/- 0.70 mm with the Colvard pupillometer and 0.01 +/- 0.20 mm with the digital photograph image. The digital photograph image showed better agreement. The coefficient of interrater repeatability was smaller for the digital photograph image (0.39) than for the Colvard pupillometer (0.70). CONCLUSIONS: Scotopic pupil measurement using an infrared digital camera with a burst shot had good agreement with the Colvard pupillometer and better repeatability. The infrared digital camera is less expensive, and pupil unrest can be overcome by taking serial images. PMID- 17137994 TI - Four corneal presbyopia corrections: simulations of optical consequences on retinal image quality. AB - PURPOSE: To investigate the possibility of multifocal or aspherical treatment of the cornea with optical ray tracing. SETTING: Institute for Refractive and Ophthalmic Surgery, Zurich, Switzerland. METHODS: The optical consequences of 4 corneal shapes-global optimum (GO) for curvature and asphericity, central steep island (CSI), decentered steep island (DSI), and centered steep annulus (CSA)-for presbyopia correction were analyzed using a modified Liou-Brennan eye model and ray tracing with a commercial optic design software (Zemax, Zemax Development Corp.). The ocular optical configuration for far vision was a point light source at a distance of 5 m, 1 degree up, and a pupil diameter of 5.0 mm and for near vision, 0.4 m distance, 1 degree up, and a pupil diameter of 2.5 mm. The curvature radius (R) of the cornea and its asphericity (Q) were used as operands to optimize (simultaneously for near and far vision) the quality of the retinal image described by means of the minimum spot diameter or the root-mean-square (RMS) wavefront error. RESULTS: Starting from an emmetropic eye optimized for R and Q, the RMS wavefront error in the retina was 0.07 microm (far) and 1.42 microm (near). The GO resulted in a wavefront error of 1.42 microm (far) and 0.52 microm (near); improvement of near vision using reading glasses is possible. The CSI yielded 0.91 microm (far) and 0.13 microm (near); spectacles did not improve far or near vision. The DSI and CSA had significantly worse results for near and far vision. CONCLUSIONS: Of the options studied, GO and CSI seemed the most promising alternatives for corneal presbyopia correction. Although reading glasses can improve near vision in GO, reading glasses did not improve near vision in CSI-treated eyes. The CSI treatment is critically dependent on centration and a reverse treatment is difficult to achieve. PMID- 17137995 TI - 2005 ESCRS Ridley Medal Lecture: will keratectasia be a major complication for LASIK in the long term? AB - The literature on keratoconus and progressive post-laser in situ keratomileusis ectasia (PPLK) was studied after a retrospective review of 137 highly myopic eyes (mean -15.58 diopters) 5 to 9 years after LASIK. Only 2 eyes developed keratectasia, 1 after automated lamellar keratolasty for myopia and 1 after myopic LASIK. In neither case was intraoperative pachymetry carried out. In the PPLK case, a residual stromal bed thickness (RSBT) of 120 microm was discovered when a retreatment was attempted on lifting the flap. The PPLK developed 2 years later. The obvious discrepancy between reported cases of PPLK and the expected rate from the incidences in larger retrospective studies and our own 15-year experience suggests that PPLK is not a significant complication provided the standard rules applying to the preoperative total corneal thickness and RSBT are observed. However, newer techniques for the study of posterior corneal changes and those involving corneal biomechanics should be used in the future to study the long-term effects of LASIK on the cornea. PMID- 17137996 TI - Traumatic pupillary capture of the haptic of an angle-supported phakic intraocular lens. AB - A 27-year-old woman had implantation of an angle-supported phakic intraocular lens (pIOL) for myopia in the left eye in 2003. Moderate iris atrophy and pupil ovalization were noted in 2005. In 2006, the eye was hit by a plastic bullet shot from a toy gun. The pupil became distorted as the inferior haptic engaged the pupil edge. Visual acuity was not affected. Surgical repositioning was uneventful, yielding a round pupil with no damage to the iris sphincter or the lens. The endothelium was not affected by the trauma or surgery. Previous iris atrophy may have facilitated pupillary capture because of reduced iris elasticity and pupil ovalization by the haptic. Blunt trauma can dislocate angle-supported pIOLs. Implantation of these IOLs should be discouraged in patients who perform activities that put them at risk for eye trauma. PMID- 17137997 TI - Intraocular ophthalmic ointment following clear corneal phacoemulsification: Clinical implications. AB - We report 4 cases of apparent ophthalmic ointment in the anterior chamber after sutureless clear corneal phacoemulsification and posterior chamber intraocular lens implantation. The cases, as well as previous literature, indicate that ointment for topical use can be well tolerated in the eye, although glaucoma and uveitis can be potential negative outcomes. Possible risk factors, some of which may be related to current rates of endophthalmitis after clear corneal cataract surgery, and methods to prevent intraocular ophthalmic ointment after cataract surgery are discussed. PMID- 17137998 TI - Effect of circular keratotomy on keratoconus. AB - Circular keratotomy was performed in 3 eyes of 3 patients with keratoconus before planned lamellar keratoplasty. After peribulbar anesthesia, a circular 400 microm cut centered on the geometrical corneal center was made using a Hessburg-Barron 7.0 mm trephine (Altomed). The circular cut was deepened with a guarded diamond blade set at 90% of the thinnest point in the circumference and closed with a single 10-0 nylon running suture. Videokeratography revealed increased astigmatism and increased central curvature in all eyes. In 1 patient, a 0.2 improvement in best spectacle-corrected visual acuity (BSCVA) was noted, associated with overall corneal steepening. In the other 2 patients, refraction, BSCVA, and corneal profile worsened, requiring lamellar keratoplasty. Circular keratotomy increased corneal curvature and worsened keratoconus and is therefore not recommended. PMID- 17137999 TI - Polymerase chain reaction and DNA typing for diagnosis of infectious crystalline keratopathy. AB - A 63-year-old man developed infectious crystalline keratopathy (ICK) in his right eye 1 year after phacoemulsification. The white peripheral lesion was adjacent to the corneal phacoemulsification incision. Infiltrates in the form of creamy white, midstromal branching, needle-like opacities without evidence of inflammatory cells were noted. A corneal biopsy by double lamellar flap was done and studied by 3 techniques: microbiological culture, stain, and polymerase chain reaction (PCR). Fungal and bacterial PCR were positive. A second sample was necessary to obtain a positive stain and culture. The DNA sequence analysis showed Candida parapsilosis and Staphylococcus aureus as the causal agents of the crystalline keratopathy. Treatment was started with amphotericin B 1% and cefazolin 6 times a day, and systemic voriconazole was recommended. This is the first reported case of ICK after cataract surgery. Polymerase chain reaction amplification and subsequent DNA typing were useful tools in detecting and identifying the ocular pathogens involved in this case. PMID- 17138000 TI - Complications of sequential keratorefractive and vitreoretinal surgery. AB - We describe 2 complications of sequential keratorefractive and vitreoretinal surgery not previously reported. Epithelial ingrowth occurred in 1 patient who experienced laser in situ keratomileusis flap dehiscence and replacement during vitreoretinal surgery. In the second patient, a flap could not be created with a microkeratome or a femtosecond laser because of anatomical changes from previous vitreoretinal surgery. Anatomic repair of the vitreoretinal pathology and completion of keratorefractive surgery with good visual outcomes were achieved in both patients. Flap stability should be an important operative consideration for vitreoretinal surgeons; patients may have to be informed of the risk for and possible subsequent complications of iatrogenic flap dehiscence. Refractive surgeons operating on patients after vitreoretinal surgery may encounter difficulty creating a LASIK flap even with a femtosecond laser and may need to consider other keratorefractive options. PMID- 17138001 TI - Bilateral simultaneous Achromobacter xylosoxidans keratitis following penetrating keratoplasty. AB - We report a case of severe bilateral simultaneous Achromobacter xylosoxidans keratitis following penetrating keratoplasty (PKP) in a patient with graft-versus host disease. The predisposing factors, clinical features, treatment options, and final visual outcome are reviewed. To our knowledge, this is the first report of bilateral A xylosoxidans keratitis following PKP. PMID- 17138002 TI - Superficial keratectomy after epi-LASIK. PMID- 17138003 TI - Trypan-blue-assisted "re-rhexis" for smooth in-the-bag exchange of a calcified intraocular lens. PMID- 17138004 TI - Standing phacoemulsification: a national survey. PMID- 17138005 TI - Can spasticity and dystonia be independently measured in cerebral palsy? AB - Selecting and evaluating appropriate treatments for children with cerebral palsy has been challenging. One difficulty is in the ability to quantify the presence and importance of coexisting motor signs. This study presents quantitative measures developed to assess spasticity and dystonia. Children diagnosed with extrapyramidal or spastic cerebral palsy and matched control children were studied. Spasticity was measured as the slope of the force-velocity relationship from a test where we measured the forces required to passively extend the elbow at different velocities. Dystonia was assessed by measuring "overflow" movements of arm during active movement of the other arm. Measures of dystonia and spasticity did not correlate with one another, but did correlate with their respective clinical measurement tools, the Modified Ashworth scale and the Barry Albright Dystonia scale. Most children had a combination of both spasticity and dystonia, despite diagnosis. Our measures also related to different aspects of reaching: children with increased dystonia made more curved paths, and children with increased spasticity hit higher peak velocities. These measurements allow us to distinguish between different motor disorders and the degree to which each contributes to reaching performance. Use of quantitative measures should improve selection and evaluation of treatments for childhood motor disorders. PMID- 17138006 TI - Total antioxidant/oxidant status in meningism and meningitis. AB - The objective of this study was to investigate the antioxidant/oxidant status of serum and cerebrospinal fluid in children with meningismus and acute bacterial meningitis. Twenty-three children (age range, 0.75 to 9 years) with fever and meningeal signs that required analysis of the cerebrospinal fluid, but no cytologic or biochemical evidence of meningitis in their serum and cerebrospinal fluid, constituted the meningismus group. Thirty-one children (age range, 0.5 to 10 years) with acute bacterial meningitis constituted the meningitis group. Twenty-nine healthy children (age range, 0.5 to 11 years) were recruited as control subjects. Antioxidant status (ascorbic acid, albumin, thiol, uric acid, total bilirubin, total antioxidant capacity, catalase and ceruloplasmin concentrations) and oxidant status (lipid hydroperoxide and total oxidant status) were measured. The serum antioxidant status was lower, and oxidant status levels higher in both meningitis and meningismus subjects than in the control children (P < 0.001). Cerebrospinal fluid oxidant status was lower in the meningitis group than in the meningismus group (P < 0.05). These results indicate that serum antioxidant status was lower, and serum oxidant status was higher in children in the meningismus and meningitis groups, whereas cerebrospinal fluid oxidant status was higher in the meningismus group than in the meningitis group. PMID- 17138007 TI - Links between abnormal brain structure and cognition in holoprosencephaly. AB - Converging information on medical issues, motor ability, and cognitive outcomes is essential when addressing long-term clinical management in children with holoprosencephaly. This study considered whether adding more informative structural indices to classic holoprosencephaly categories would increase prediction of cognitive outcomes. Forty-two children with holoprosencephaly were examined to determine the association of deep gray nuclei abnormalities with cognitive abilities and the effect of motor skill deficits on cognitive performance. Additionally, a cognitive profile was described using the Carter Neurocognitive Assessment, an experimental diagnostic instrument designed specifically for young children with severe neurodevelopmental dysfunction. Findings indicated that nonseparation of the deep gray nuclei was significantly associated with the cognitive construct of vocal communication, but not with the cognitive constructs of social awareness, visual attention, or auditory comprehension. Importantly, motor skill deficits did not significantly affect performance on the Carter Neurocognitive Assessment. This study is the first investigation to provide a descriptive overview of specific cognitive skills in this group of children. The results also strongly suggest that this feature of the brain's structure does not predict all aspects of neurodevelopmental function. These findings contribute a critical component to the growing body of knowledge regarding the medical and clinical outcomes of children with holoprosencephaly. PMID- 17138008 TI - Influenza A and febrile seizures in childhood. AB - The aims of the present study are to identify predisposing factors of febrile seizures in influenza A infection and to clarify the special characteristics of febrile seizures in children with influenza A infection. Between January and July 2005, children hospitalized because of febrile seizures and subsequently confirmed influenza A infection were enrolled as subjects. Age-matched control subjects were those admitted as a result of influenza A infection but no febrile seizures (control 1) and children who developed febrile seizures with negative viral studies (control 2). Significant factors for the development of febrile seizures include: history of febrile seizures, family history of seizure disorders, and coexisting gastroenteritis. Independent risk factor for febrile seizures was history of febrile seizures (odds ratio 7.58, 95% confidence interval CI 1.48 to 38.84, P = 0.015). When compared with children who developed febrile seizures with negative virus studies, children who developed febrile seizures in influenza A infection had a significantly higher maximum body temperature, shorter duration of fever before seizure onset, and more frequent occurrence of partial seizures. Current episode represented first seizure in 26.5% of children infected with influenza A as compared with 50% of children whose virus studies were negative (P = 0.04). The findings suggest that effective vaccination may prevent development of febrile seizures, especially in those patients with past history of febrile seizures. Rapid diagnostic testing for influenza infection in the management of complex febrile seizures, especially during influenza season, is cost-effective. PMID- 17138009 TI - Evaluation of a multimodal management of prematurity-related spasticity. AB - To examine the efficacy of a rehabilitation protocol, focusing on spasticity management through botulinum toxin A injections in the lower limbs, an etiologically homogeneous group of 57 prematurely born children with cerebral palsy was prospectively evaluated (minimum follow-up 18 months) under pragmatic conditions. Gross Motor Function Classification System categories were: I = 12, II = 9, III = 16, IV = 15, V = 4. Outcome was evaluated with goniometry, Gross Motor Function Measure, functional goal attainment at baseline and in subsequent months, the Gross Motor Function Classification System, functional mobility status, and parents' satisfaction at more than 18 months after first botulinum toxin. Goniometry demonstrated significantly improved range of movement in lower limbs at 10 days and 1 month after botulinum toxin. Differences persisted >18 months at the popliteal angles (P < 0.001). Gross Motor Function Measure changed significantly in 20 children (8 points in total score) at 3 months after first botulinum toxin (P < 0.0001) with less significant results thereafter. Predetermined functional goals were achieved in 61% at >18 months. Parents were satisfied in approximately 90% of the cases. Eighteen of 57 children (31.57%) changed Gross Motor Function Classification System status over a mean of 33.8 months (18-48) follow-up. Most significant gains were recorded in the severely involved group IV, where 10 of 15 (66.66%) improved. The high percentage of change in group IV implies the importance of gained sitting balance due to spasticity management. PMID- 17138010 TI - Communicating a diagnosis of cerebral palsy: caregiver satisfaction and stress. AB - As part of the implementation of a population-based registry of children with cerebral palsy, caregiver satisfaction with the process by which diagnosis was originally communicated by a professional was assessed. Satisfaction with various aspects of the diagnosis process was assessed using a five-point Likert scale and related to child, family, and situational characteristics. Measures were then correlated with current caregiver stress as measured objectively by the Parenting Stress Inventory. During the registration process, 59 consecutive caregivers (55 mothers) were questioned. Overall, 62% (35/59) were satisfied with the disclosure process, with satisfaction ranging from 69% (41/59-hopefulness) to 92% (54/59 honesty) for professional qualities, and from 61% (36/59-sufficient information provided) to 78% (46/59-understandable) for disclosure content. Satisfaction was related to the quantity and content of information given at the disclosure session. Parenting Stress Inventory scores, both total and for parental distress, correlated significantly with both the severity of the child's cerebral palsy and caregiver satisfaction with varying elements of the disclosure process. Overall caregiver satisfaction with the process by which a diagnosis of cerebral palsy is given appears to be good. Together with the severity of a child's intrinsic cerebral palsy, it appears to relate to later parental adjustment to a setting of chronic disability, suggesting a portal through which improvements in information delivery may result in better familial adaptation to disability. PMID- 17138011 TI - Short-duration ACTH therapy for cryptogenic West syndrome with better outcome. AB - In this study, seven patients with cryptogenic West syndrome were subjected to short-duration adrenocorticotrophic hormone (ACTH) therapy. ACTH was administered daily for 7 to 12 days (mean 10 days), and then was withdrawn. Daily single dose was 0.022 to 0.027 mg/kg/day (0.024 mg/kg/day, 0.96 IU/kg), total dose was 0.17 to 0.28 mg/kg (0.23 mg/kg, 9.0 IU/kg), and treatment lag was 12 to 105 days (median 14 days). ACTH therapy controlled tonic spasms with no serious side effects in all patients, but two patients still had electroencephalographic abnormality. The intelligence quotients or developmental quotients of six patients with short treatment lag were 79 to 110 at the age of 2 to 6 years; only one patient with long treatment lag had a developmental quotient of 60. This new short-duration ACTH therapy could yield better cognitive outcomes for cryptogenic West syndrome. PMID- 17138012 TI - The neurologic aspects of PHACE: case report and review of the literature. AB - PHACE syndrome is a neurocutaneous disorder characterized by large cervicofacial infantile hemangiomas and associated anomalies of the brain, cerebrovasculature, aorta, heart, and eyes. Two categories of neurologic disease are observed among PHACE patients: congenital malformations of the cerebellum, cerebrum, and cerebral vasculature and progressive stenoses and occlusions of principal cerebral arteries. A subgroup of patients develops a moyamoya-like vasculopathy and consequent ischemic strokes. This report details the clinical course and management of a young female with PHACE and reviews the various neurologic aspects of this neurocutaneous disorder. This patient presented with high-grade stenoses of the internal carotid arteries bilaterally, formation of extensive vascular collaterals, and multiple ischemic strokes. She underwent bilateral pial synangiosis procedures and has not had stroke recurrence in 2 years of follow-up. The presence of a characteristic infantile hemangioma necessitates further evaluation for the extracutaneous features of PHACE. In cases of steno-occlusive vasculopathy, we recommend early consideration of encephaloduroarteriosynangiosis or a commensurate revascularization procedure. PMID- 17138013 TI - Hereditary hypotonia, muscle weakness, failure to thrive, and cognitive delay in a large moslem kindred. AB - Five infants of a Moslem-Arab extended family were evaluated for common and characteristic clinical findings of failure to thrive, extreme muscle weakness, severe motor delay, and moderate to severe cognitive and verbal delay. All children were below the third percentile in weight and height, and three of them had head circumference below the third percentile. Neurologic examination revealed severe hypotonia, muscle weakness, and absent deep tendon reflexes. Two children died at 2 years of age, and none of the children acquired full head control and the motor milestones of rolling and sitting. Laboratory evaluation including muscle biopsies, genetic studies, and metabolic evaluation was nondiagnostic. PMID- 17138014 TI - Acute myelitis in a child: current hypotheses. AB - This report presents the case of a child with atopic dermatitis, who developed progressive muscular weakness and hypotonia of the four limbs. The cervical spinal cord magnetic resonance imaging revealed a C(4) lesion (T(2)-weighted images); the cerebrospinal fluid findings were normal. Treatment with intravenous immunoglobulins and methylprednisolone obtained rapid clinical improvement, and approximately 1 month later the small C(4) lesion disappeared. Various diagnostic hypotheses are discussed: acute myelitis by infective agents, acute disseminated encephalomyelitis, multiphasic disseminated encephalomyelitis, multiple sclerosis, and isolated postinfective myelitis. Another hypothesis relates to atopic myelitis, a form recently described in the Japanese literature, associated with atopic dermatitis, hyperIgEemia, and high levels of specific immunoglobulin E to Dermatophagoides farinae and Dermatophagoides pteronyssinus. This diagnosis is difficult to confirm without biopsy evidence of eosinophilic inflammation. PMID- 17138015 TI - Decreased cortical excitability due to a large venous angioma. AB - This study reports on a patient with episodes of right hand paralysis and complete sensory loss, considered to be functional because of a glove-like distribution of the sensory deficit, normal motor and sensory nerve conduction studies of median and ulnar nerve as well as normal median nerve somatosensory evoked potentials. Transcranial magnetic stimulation indicated increased threshold of the left hemisphere. Neuroimaging studies revealed a large venous angioma in the left frontal lobe. PMID- 17138016 TI - Causality of pediatric brainstem infarction and basilar artery fenestration? AB - Pediatric arterial thromboembolic stroke is an uncommon condition and rarely is reported to be associated with a cerebral artery fenestration. This clinical report discusses the case of a child with brainstem infarction and basilar artery fenestration. A cardiac source of thromboembolic events could be excluded; however, detailed coagulation analysis revealed in addition an apoliopoprotein(a) size polymorphism. Because we assume that the two concurrent pathologies in combination caused the arterial thromboembolic stroke, the evaluation of all potential triggers including vascular anomalies and coagulation disorders should be considered in unexplained pediatric infarction. PMID- 17138017 TI - Bithalamic lesions of butane encephalopathy. AB - Butane inhalation can cause serious medical complications and is particularly toxic to the nervous system. This is a report of an acutely encephalopathic youth with prominent abulia. MRI revealed severe bithalamic injury attributed to butane toxicity. Clinical issues, including particular radiologic findings, related to butane inhalation are reviewed. PMID- 17138018 TI - Moyamoya syndrome in a child with trisomy 12p syndrome. AB - A female, 2 years and 7 months of age, was admitted to the hospital with stupor and nystagmus following projectile vomiting. She had been prenatally diagnosed with trisomy 12p with a familial pericentric inversion of chromosome 12 originating from her mother. She manifested developmental delay and some dysmorphic features of the face and limbs compatible with the clinical features of trisomy 12p. Four-vessel cerebral angiography revealed severe stenosis and occlusion of the supraclinoid portion of the right and left internal carotid arteries with numerous collateral vessels in the vicinity of the occlusion. These features are consistent with moyamoya syndrome. This report presents the first case of moyamoya syndrome with trisomy 12p with a familial pericentric inversion of chromosome 12. PMID- 17138019 TI - Botulinum toxin type A in the management of cerebral palsy: low or high dose? PMID- 17138020 TI - Oligohydrosis and topiramate. PMID- 17138022 TI - President's message update from advocacy committee. PMID- 17138024 TI - Guidelines and standards for performance of a pediatric echocardiogram: a report from the Task Force of the Pediatric Council of the American Society of Echocardiography. PMID- 17138025 TI - Assessment of the evolution of normal fetal diastolic function during mid and late gestation by spectral Doppler tissue echocardiography. AB - OBJECTIVE: To establish gestational age-specific reference values in healthy singleton fetuses, we prospectively assessed the evolution of diastolic longitudinal wall-motion velocities by spectral Doppler tissue imaging. METHODS: Early (Ea) and late diastolic (Aa) peak Doppler tissue imaging velocities were analyzed in 114 fetuses (age range: 14-42 weeks) at the base of right ventricular free wall, ventricular septum, and left ventricular free wall and compared with early (E) and late (A) diastolic peak Doppler inflow velocities. RESULTS: A linear increase in Ea, Aa, and Ea/Aa ratio was documented at all sites with advancing gestation. Likewise, the peak E flow velocities of both atrioventricular valves and the tricuspid peak A flow velocity increased. The ratio of peak E/Ea velocities decreased exponentially as a result of a more rapid increase in Ea than E, to reach a stable E/Ea relationship only in the early third trimester. CONCLUSIONS: There was a strong positive correlation between Ea and Aa velocities and gestational age indicating improved diastolic myocardial lengthening with advancing gestation. Reference charts for Doppler tissue imaging velocities were established that will allow identification of fetal diastolic function abnormalities. PMID- 17138026 TI - Left ventricular Tei index in children: comparison of tissue Doppler imaging, pulsed wave Doppler, and M-mode echocardiography normal values. AB - The Tei index has been found to be useful for analyzing systolic and diastolic global ventricular function in a wide variety of congenital and acquired cardiac abnormalities. However, there are some discrepancies between reports as to the normal values for the Tei index obtained by the different echocardiographic techniques and by different investigators. We conducted a prospective study to determine the normal range of left ventricular Tei index (LVTX) values in a broad sample of children using tissue Doppler imaging, pulsed wave Doppler, and M-mode echocardiography. In all, 289 children with normal echocardiogram findings (age 1 day-18 years, body surface area 0.08-2.4 m(2), heart rate 46-182/min) were studied. The LVTX was calculated by all 3 methods in each patient during a single echocardiographic examination. The normal LVTX values (mean +/- SD) for the 3 techniques were: LVTX-Doppler tissue imaging = 0.38 +/- 0.06; LVTX-pulsed wave Doppler = 0.36 +/- 0.07; and LVTX-M-mode echocardiography = 0.29 +/- 0.08. LVTX Doppler tissue imaging and LVTX-pulsed wave Doppler values were only slightly but statistically significantly different (P < .05). LVTX-M-mode echocardiography values were consistently and significantly less than those obtained by both of the other two methods (P < .01, respectively). The effects of age, body surface area, and heart rate were not clinically significant. These results are similar but not identical to those from prior studies. PMID- 17138027 TI - Common origin of the innominate and carotid arteries: prevalence, nomenclature, and surgical implications. PMID- 17138028 TI - Asynchrony of left ventricular systolic performance after the first acute myocardial infarction in patients with narrow QRS complexes: Doppler tissue imaging study. AB - BACKGROUND: Left ventricular (LV) electromechanical delay results in asynchronized contraction. However, it is not known if the presence of cardiac diseases without QRS prolongation may result in interventricular or intraventricular asynchrony. Doppler tissue imaging is now established for detecting regional contractile abnormalities and asynchrony in the LV. OBJECTIVES: The aim of the study was to assess the degree of LV asynchrony after the first acute myocardial infarction (AMI) in patients with a narrow QRS complex using Doppler tissue imaging and correlate this with the site and extent of the infarction. METHODS: Echocardiography with Doppler tissue imaging was performed within 1 week of AMI in 155 patients and compared with 50 age- and sex-matched healthy volunteers. Regional myocardial velocities were assessed at the 4 mitral annular sites, and the corresponding systolic velocity (Sm), early diastolic velocity (Em), time to peak Sm (Ts), and time to peak Em (Te) were measured. To assess LV synchronicity, SDs of Ts (Ts-SD) and Te (Te-SD) of all the 4 mitral annular sites were computed. Location and size of infarct were confirmed by echocardiographic wall-motion score index. RESULTS: QRS complex duration was normal in all patients. Wall-motion score index was significantly higher in patients with anterior than inferior AMI (2.02 +/- 0.34 vs 1.24 +/- 0.21, P < .001). Ts-SD was significantly higher in patient than control group, and in patients with anterior than inferior AMI (38.21 +/- 2.59 vs 21.06 +/- 0.52 milliseconds and 43.18 +/- 3.77 vs 33.24 +/- 1.4 milliseconds, respectively, P < .001 for each), whereas Te-SD did not differ significantly among these groups (20.35 +/- 1.77 vs 18.17 +/- 1.14 milliseconds and 21.6 +/- 1.35 vs 19.1 +/- 1.11 milliseconds, respectively, P > .05 for each). A strong positive correlation was detected between LV systolic asynchrony (Ts-SD) and wall-motion score index (r = .77), LV mass (r = .67), LV end-systolic dimension (r = .65), and LV end diastolic dimension (r = .5). The correlation was negative with LV ejection fraction (r = -.70) and Sm (r = -.6); the correlation was weak with Em (r = -.33) (P < .001 for all). In multivariate logistic regression analysis, infarct size was found to be the most independent predictor for systolic asynchrony (odds ratio 3.59, 95% confidence interval [1.43-9.33], P < .001). CONCLUSION: AMI has a significant impact on regional myocardial contractility and LV systolic (but not diastolic) synchronicity early in the course even in the absence of QRS widening or bundle branch block. The degree of LV systolic asynchrony is greater with anterior than inferior AMI and mainly determined by infarct size. PMID- 17138029 TI - High prevalence of clinically silent severe mitral regurgitation in patients with heart failure: role for echocardiography. AB - Mitral regurgitation (MR) is common in patients with congestive heart failure (CHF) and has adverse effects on prognosis. In view of the altered hemodynamics in CHF, we examined the accuracy of auscultation for its detection in CHF with reduced systolic function. We examined 370 patients on stable heart failure therapy enrolled in our CHF clinic, if they had a physical examination by one of the two senior cardiologists and an echocardiogram within 2 weeks after the physical examination. The MR murmur was graded 0 to 6 on physical examination. MR was graded 0 to 4 on echocardiography using standard echocardiographic criteria. The left ventricular (LV) ejection fraction was 21 +/- 12% and age was 65 +/- 15 years. By echocardiography MR was present in 345 patients (94%), being 1+ in 162 (44%), 2+ in 80 (22%), 3+ in 54 (15%), and 4+ in 51 (14%). The frequency of an audible MR murmur by physical examination was 4% in 1+ MR, 11% in 2+ MR, 13% in 3+ MR, and 37% in 4+ MR. The audibility of MR murmur was not related to age, sex, body size, LV size, ejection fraction, or left atrial size. In conclusion, grade 3 or 4+ MR is present in one third of patients with CHF and reduced ejection fraction, and is inaudible in three quarters of them by physical examination. Echocardiography is needed for its detection. Audibility of MR murmur cannot be predicted by body size, LV size, or LV function in these patients. PMID- 17138030 TI - Comparison of regional versus global assessment of left ventricular function in patients with left ventricular dysfunction, heart failure, or both after myocardial infarction: the valsartan in acute myocardial infarction echocardiographic study. AB - BACKGROUND: Left ventricular (LV) ejection fraction (EF) and wall-motion index (WMI) have both been shown to be independent predictors of outcome after myocardial infarction (MI). OBJECTIVES: We sought to determine whether these two measurements of LV systolic function provide similar or complementary information about prognosis after MI. METHODS: Echocardiography was performed in 610 patients with LV dysfunction, heart failure, or both after MI enrolled in the Valsartan in Acute MI trial. LVEF was estimated by biplane Simpson's rule, and WMI was assessed using a 16-segment model in 502 patients with echocardiograms of sufficient quality for wall-motion assessment. RESULTS: Both LVEF and WMI were independent predictors of adverse outcome after MI. LVEF conferred no additional prognostic information in multivariable analysis including WMI (P = .39) or number of affected segments (P = .53), whereas WMI (P = .02) and total number of affected segments (P = .006) remained significant even when adjusting for LVEF. CONCLUSIONS: Assessment of regional dysfunction by WMI or the number of affected segments has slightly more prognostic value than LVEF in patients with LV dysfunction, heart failure, or both after MI. Regional assessment might be a more sensitive predictor of outcome than global assessment in patients with acute MI. PMID- 17138031 TI - Reduced myocardial contractility assessed by tissue Doppler echocardiography is associated with increased risk during adrenal surgery of patients with pheochromocytoma: report of a preliminary study. AB - BACKGROUND: Depressed myocardial contractility, although rarely reported in pheochromocytoma, might be underestimated. It may be a determinant of perioperative risk during adrenal surgery. METHODS: We prospectively studied consecutive patients with pheochromocytoma; myocardial function examined by standard and tissue Doppler echocardiography was compared with matched control subjects. The incidence of hemodynamic collapse during adrenal surgery was measured. RESULTS: A total of 15 patients were included (8 men, 46 [17] years, hypertension in 10). All but one had a normal left ventricular ejection fraction. However, compared with control subjects, they had a depressed systolic strain rate (SR) (1.8 [2.1] vs 4.1 [2.2] s(-1), P = .007). Furthermore, 6 of 8 patients with systolic SR less than 2 s(-1) experienced intraoperative collapse, versus 1 of 7 with SR greater than 2 s(-1) (P = .041). No association was observed with other variables. CONCLUSIONS: Patients with pheochromocytoma may have depressed myocardial contractility detected by tissue Doppler echocardiography despite a normal standard echocardiogram. A systolic SR less than 2 s(-1) was associated with an increased risk of perioperative collapse. PMID- 17138032 TI - Impact of type 2 diabetes mellitus on aortic elastic properties in normotensive diabetes: Doppler tissue imaging study. AB - OBJECTIVES: The stiffening of aorta and other central arteries is a potential risk factor for increased cardiovascular morbidity and mortality. The association of hypertension with type 2 diabetes may obscure the degree to which diabetes alone contributes to impaired arterial function. This study examined whether the presence of type 2 diabetes alone is associated with an impaired aortic mechanical function in patients with or without coronary artery disease (CAD). METHODS: In all, 154 patients were recruited and assigned to groups A (n = 46, type 2 diabetes with no CAD), B (n = 64, nondiabetic CAD), or C (n = 44, diabetes with CAD) and 20 age- and sex-matched healthy participants were enrolled in a control group. Patients were recruited from those sent for coronary angiography. CAD was excluded for group A. Pulse pressure, aortic strain, and distensibility were calculated from the aortic diameters measured by echocardiography and blood pressure obtained by sphygmomanometer. Aortic wall systolic velocity was measured using pulsed wave Doppler tissue imaging. RESULTS: Pulse pressure was significantly higher in patient groups A, B, and C in comparison with control group (40.2 +/- 9, 40.1 +/- 11, and 50.2 +/- 13 vs 35.5 +/- 9 mm Hg [P < .01], respectively). The pulsatile change in the aortic diameter and distensibility were less in the patient groups than in the control group (11 +/- 4%, 8 +/- 5%, and 8 +/- 4% vs 17 +/- 9% [P<.001], and 6 +/- 2, 6 +/- 1, and 3 +/- 2 vs 10 cm(2)/dyne/10(3), respectively). In addition, the aortic wall systolic velocity was significantly lower in patient groups compared with control group (6 +/- 2, 6.1 +/- 1, and 5.1 +/- 1 vs 8.5 +/- 1.5 cm/s [P < .01], respectively). Although aortic function parameters were very declined for group C, there was no significant difference between groups A and B that reflected equivalent risk. In diabetic groups A and C, aortic strain, distensibility, and aortic wall systolic velocity showed strong negative correlation with the duration of diabetes (r = .53, r = -.68, and r = -.56, respectively) and glycosylated hemoglobin (HBA(1)) (r=-.64 [P < .01], r = -.77 [P < .001], and r = -.57 [P < .01], respectively). CONCLUSION: The increased aortic stiffness that affects patients with type 2 diabetes seems to be an early event that may explain why patients with diabetes have a particularly high risk of developing cardiovascular complications. Poor glycemic control and duration have detrimental effect on aortic elastic properties. PMID- 17138033 TI - Noninvasive diagnosis of coronary artery bypass graft disease by dobutamine stress real-time myocardial contrast perfusion imaging. AB - BACKGROUND: The aim of this study was to assess the accuracy of real-time myocardial contrast perfusion imaging (MCPI) during dobutamine stress echocardiography, for the diagnosis of coronary artery bypass graft disease. METHODS: MCPI was performed using commercially available ultrasound contrast agents (Optison or Definity) at rest and at peak dobutamine-atropine stress in 64 patients with previous coronary artery bypass graft. Significant disease was defined as 70% or greater stenosis in one or more grafts or a native nongrafted coronary artery. MCPI was considered diagnostic in the presence of reversible perfusion abnormalities (RPA). RESULTS: Significant stenosis (>or=70% by quantitative angiography) in one or more grafts was detected in 49 patients (77%). RPA were detected in 44 of these patients (sensitivity 90%, confidence interval [CI] 81-98). Significant stenosis was detected in 74 of the 176 bypass grafts (42%). RPA were detected in the distribution of 55 diseased grafts and 21 nondiseased grafts (regional sensitivity 74%, CI 64-84; specificity 79%, CI 71 87; and accuracy 77%, CI 71-84). RPA were detected in two or more vascular distributions in 25 of 33 patients with multivessel stenotic lesions and in 4 of 31 patients without (sensitivity 76%, CI 61-90; specificity 87%, CI 75-99; and accuracy 81%, CI 72-91). CONCLUSION: Dobutamine stress MCPI is a useful technique for the evaluation of coronary artery bypass graft disease. PMID- 17138034 TI - Effect of time delay on the diagnostic use of contrast echocardiography in patients presenting to the emergency department with chest pain and no S-T segment elevation. AB - BACKGROUND: The purpose of this study was to assess the effect of time delay on the diagnostic and prognostic use of contrast echocardiography (CE) in patients presenting to the emergency department (ED) with chest pain (CP) and no S-T segment elevation. METHODS: Patients (n = 957, 498 men) presenting to the ED within 12 hours of suggested cardiac CP underwent CE. Regional function (RF) and myocardial perfusion were interpreted separately by expert readers blinded to all other clinical data. Primary (acute myocardial infarction and total mortality) and secondary (unstable angina and revascularization) events within 24 hours of enrollment were determined. RESULTS: Patients were divided into 4 quartiles based on the time interval between their last episode of CP and CE. Patients in quartile I had CE during CP (time delay of 0 minutes). The time delay in quartiles II, III, and IV were 54 +/- 45, 213+/-54, and 556 +/- 184 minutes, respectively (P < .001). The incidence of events was similar among the 4 quartiles. In each quartile, patients with normal RF had the lowest incidence of events, whereas those with both abnormal RF and myocardial perfusion had the highest incidence of events. Patients with abnormal RF but normal myocardial perfusion had an intermediate event rate. CONCLUSIONS: In patients presenting to the ED within 12 hours of CP, the timing of CE does not influence its ability to predict events that occur 24 hours later. These findings have important implications in the performance of CE in the ED. PMID- 17138035 TI - Real-time simultaneous triplane contrast echocardiography gives rapid, accurate, and reproducible assessment of left ventricular volumes and ejection fraction: a comparison with magnetic resonance imaging. AB - OBJECTIVE: We sought to compare the feasibility, accuracy, and reproducibility of simultaneous triplane echocardiography for measurements of left ventricular (LV) volumes and ejection fraction (EF) with reference to magnetic resonance imaging (MRI). METHODS: Digital echocardiography recordings of apical LV views with and without intravenous contrast were collected from 53 consecutive patients with conventional 2-dimensional (2D) imaging and with simultaneous triplane imaging. MRI of multiple LV short-axis sections was performed with a 1.5-T scanner. Endocardial borders were manually traced, and LV volumes and EF from 2D biplane echocardiography and MRI were calculated by method of disks. On triplane data, a triangular mesh was constructed by 3-dimensional interpolation and volumes calculated by the divergence theorem. RESULTS: Triplane image acquisition was less time-consuming than 2D biplane. Precontrast feasibility was 72% for triplane and 82% for 2D biplane images, increasing to 98% and 100% with contrast, respectively. Bland-Altman analysis demonstrated LV volume underestimation by echocardiography versus MRI, which was significantly reduced by contrast and triplane imaging. The 95% limits of agreement for EF between echocardiography and MRI narrowed using triplane compared with 2D biplane (precontrast -12.5 to 6.7% vs -17.2 to 9.9%, and with contrast -7.1 to 5.8% vs -9.4 to 6.4%, respectively). At intraobserver and interobserver analysis of 20 patients, limits of agreement for EF narrowed with contrast triplane compared with 2D biplane. CONCLUSION: Simultaneous LV triplane imaging is feasible with simple and rapid image acquisition and volume analysis, and with contrast enhancement it gives accurate and reproducible LV EF measurements compared with MRI. PMID- 17138036 TI - Three-dimensional echocardiography improves the understanding of the mechanisms and site of left atrioventricular valve regurgitation in atrioventricular septal defect. AB - OBJECTIVES: The purpose of this study was to determine whether 3-dimensional echocardiography (3DE) provides additional information regarding the mechanisms and sites of left atrioventricular valve regurgitation in atrioventricular septal defect compared with transesophageal 2-dimensional echocardiography (2DE). METHODS: Eleven patients with a median age of 5.4 years (2.9-11.6 years) and a median weight of 16.8 kg (13.7-38.3 kg) with an atrioventricular septal defect underwent simultaneous transesophageal 2DE and 3DE before operation. RESULTS: The 2DE-3DE agreement for the assessment of the superior and mural leaflet size was 72.7%. The 2DE-3DE agreement for coaptation failure, a residual or primary cleft, and commissural abnormalities as a mechanism of regurgitation were 72.7%, 63.6%, and 36.4%, respectively. For jet sites the 2DE-3DE agreement was 63.6% for a commissural and central location. CONCLUSION: Three-dimensional echocardiography provides new and superior data regarding the mechanisms and sites of left atrioventricular valve regurgitation in atrioventricular septal defect. PMID- 17138037 TI - Illustration of the additional value of real-time 3-dimensional echocardiography to conventional transthoracic and transesophageal 2-dimensional echocardiography in imaging muscular ventricular septal defects: does this have any impact on individual patient treatment? AB - OBJECTIVE: We sought to answer the question of whether the additional morphologic details obtained by real-time 3-dimensional (3D) echocardiographic (RT3DE) imaging of muscular ventricular septal defect (VSD) has any significant impact on treatment options of individual patient. BACKGROUND: Muscular VSD can be safely and effectively closed by interventional catheterization procedure using VSD devices under transesophageal echocardiographic (TEE) guidance. Recent application of RT3DE has shown great promise for imaging VSD with better display of the exact geometry, size, and location of the defect. METHODS: Nineteen patients with different types of VSDs were imaged with RT3DE matrix-array transducer; there were 6 cases with muscular VSD. Based on standard transthoracic echocardiographic and TEE imaging, one patient was considered a good candidate for perventricular VSD device occlusion, three patients were considered for surgical closure, and in two patients no intervention was deemed necessary. RESULTS: RT3DE successfully displayed the exact morphology of the VSD in all 6 patients, whereas transthoracic echocardiography and TEE showed the defect as a dropout with variable diameter in different views. Such planer images did not accurately predict the exact morphology in the patient in whom device occlusion was considered and the device embolized to the left ventricle in a few heartbeats. Surgical circular patch was used in two patients and primary suture was used in two patients in agreement with the 3D morphology. In two patients the 3D morphology of the VSD was small enough that no intervention was considered. CONCLUSIONS: RT3DE imaging of muscular VSD can accurately display the exact geometry of the defect, which can have significant impact on treatment strategies of individual patients. This new imaging modality should be an important adjunct to the standard transthoracic echocardiographic and TEE imaging of these defect before any intervention. PMID- 17138038 TI - Impact of anesthesia on echocardiographic evaluation of systolic and diastolic function in rats. AB - BACKGROUND: Echocardiography is used on rats but general anesthesia is usually necessary to be able to obtain a good quality echocardiogram. Each type of anesthetic agent has specific impacts on hemodynamics and, therefore, may affect differentially the echocardiographic measurements. OBJECTIVES: We sought to compare the echocardiograms of normal rats and rats with chronic aortic regurgitation under anesthesia using ketamine-xylazine or isoflurane. METHODS: Animals underwent an echocardiogram with both drugs sequentially. Echocardiographic measurements were compared. RESULTS: Mitral diastolic Doppler measurements (early diastolic filling wave [E] and late atrial diastolic filling wave [A] velocities) were significantly affected by the type of anesthesia in the normal group but not left ventricular dimensions or ejection fraction. Left ventricular dimensions were affected by the type of anesthesia in the aortic regurgitation group and diastolic Doppler flow. CONCLUSION: The anesthetic agent has significant specific impacts on many echocardiographic measurements. Investigators working with rat models should be aware of those potential effects. PMID- 17138039 TI - The architecture of a mobile outreach echocardiography service. AB - We describe the outreach echocardiography program at our tertiary care referral center in southeast Minnesota. Cardiac sonographers from our institution transport ultrasonographic imaging equipment to regional hospitals and clinics where they perform complete transthoracic echocardiographic examinations. Digital images from standard 2-dimensional, color flow, and Doppler echocardiography are transferred to our clinic for interpretation by a cardiologist. The outreach program enables physicians without access to echocardiography to use this powerful diagnostic tool in the local evaluation of patients with suggested or known heart disease. We detail the organization of our outreach echocardiography program and emphasize the team approach that facilitates optimal image acquisition, prompt interpretation, and timely reporting of results. PMID- 17138040 TI - Syncope with a twist. AB - Left atrial ball thrombus is an uncommon entity, seen more commonly in the presence of mitral valve disease. We present here a case of a large left atrial thrombus presenting as sudden cardiac death. PMID- 17138041 TI - Traumatic intramyocardial left ventricular dissection: a case report. AB - In this report, we present an unusual case of a patient who sustained and survived an intramyocardial dissecting hematoma with subacute ventricular rupture complicating a motor vehicle accident. In conclusion, we report on the diagnosis, management, and prior literature of this highly unusual and often lethal condition. PMID- 17138042 TI - Midventricular hypokinesis as a cardiac manifestation of anaphylaxis: a case report. AB - We report a case of anaphylaxis in which left midventricular hypokinesis was found by echocardiogram performed while the patient was hypotensive shortly after the onset of acute chest pain with S-T segment elevations. Cardiac injury was confirmed by elevation of cardiac enzymes. Repeated echocardiogram 2 days later demonstrated normal ventricular wall function and coronary angiogram demonstrated no angiographically apparent disease. We discuss possible mechanisms for cardiac injury and suggest mast cell-mediated coronary spasm as the most likely. PMID- 17138043 TI - Hematoma of a congenitally bicuspid aortic valve in a patient with polycythemia vera and the antiphospholipid antibody syndrome. AB - We present a novel case of a hematoma involving a congenitally bicuspid aortic valve. The patient was a 47-year-old woman with polycythemia vera and antiphospholipid antibody syndrome. She was treated with anagrelide for thrombocytosis. The patient developed a valvular hematoma which caused clinically significant aortic stenosis and regurgitation. Consequently, the patient underwent surgery with insertion of a porcine bioprosthesis. Eighteen months postoperatively the patient developed a large prosthetic thrombus with severe aortic stenosis requiring reoperation. PMID- 17138044 TI - Thrombotic "ghosts": echocardiographic appearance of thrombi with hollow cores and implications regarding mechanism of spontaneous clot lysis. AB - We present a case of a patient whose transesophageal echocardiogram revealed multiple thrombi in different stages of central lysis. This produced the appearance of undulating thin outer shells and lucent central cores resembling "ghosts." Although most thrombi appear to resolve by a lytic process that produces reduction in size from the exterior surface inward, the thrombi illustrated in this case appear to lyse from the interior outward. In this report we discuss the mechanisms of intracardiac clot lysis and speculate that a newly described protein factor, thrombin-mediated inhibition of fibrinolysis, may play a role in formation of hollow thrombi. PMID- 17138045 TI - Cardiac actinomycosis: an unusual cause of an intracardiac mass. AB - Actinomycosis is a chronic disease characterized by abscess formation, tissue fibrosis, and draining sinuses that may involve the cervicofacial area, thorax, abdominopelvic region, or central nervous system. We describe a patient with cardiac actinomycosis presenting with pericardial disease and an intracardiac mass. The diagnosis failed to be obtained by pericardiocentesis, but was obtained after echocardiographically guided biopsy of the intracardiac mass. The patient recovered with long-term penicillin therapy. A review of the literature highlights the frequent pericardial presentation of cardiac actinomycosis, the potential difficulty in making the diagnosis, and the remarkable clinical response and good prognosis that can result when the correct diagnosis is made and appropriate antibiotic therapy administered. PMID- 17138046 TI - Cellular cardiomyoplasty by catheter-based infusion of stem cells in clinical settings. AB - Myocardial infarction is the leading cause of congestive heart failure and death in the industrialized world. However, the intrinsic repair mechanism of the heart is inadequate. Current therapy is limited in preventing ventricular remodeling, but can not regenerate the lost cardiomyocytes. Recent interests have been focused on cellular cardiomyoplasty which is an outside intervention to support the reparative process in the heart through transplantation of stem/progenitor cells or cardiac cells. Cellular cardiomyoplasty with stem cells is a possible option to reverse the adverse hemodynamic and neurohormonal imbalance after myocardial infarction. Experimental studies and clinical trials suggest that cellular cardiomyoplasty may benefit tissue perfusion and contractile performance of the injured heart. Although the mechanisms are still intensively debated, cellular cardiomyoplasty with stem cells has already been introduced into the clinical settings. However, it is an important challenge how stem cells are delivered to targeted area. In early studies on animals, intramyocardial injection of stem cells after thoracotomy is the predominant transplantation route which is not suitable for most patients in clinical settings. Then the catheter-based infusion of stem cells is clinically introduced and rapidly developed in patients because of the safety, convenience and mini-invasion. We mainly review the progress in catheter-based transplantation with stem cells in order to fully understand the application of various intervention-based approaches to stem cells transplantation in clinical settings. PMID- 17138047 TI - The aetiology and pathogenesis of chronic allograft nephropathy. AB - Renal transplantation is the ultimate form of renal replacement therapy, and is the treatment of choice for many patients with end-stage renal failure. The advent of calcineurin inhibitor based immunosuppression resulted in the 1-year renal allograft failure rate dropping from around 50% twenty years ago to less than 10% in more recent times. Despite a massive improvement in renal allograft survival in the first year following transplantation 10-year graft survival can be as low as 50%. Chronic allograft nephropathy (CAN) is recognised as the main cause of renal allograft failure following the first year after transplantation. The diagnosis of CAN can only be made histologically. Typically biopsy specimens in grafts with CAN demonstrate an overall fibrotic appearance effecting the vascular endothelium, renal tubules, interstitium, and glomerulus. The risk factors for CAN are divided into alloimmune and alloimmune independent. Alloimmune dependent factors include acute cellular rejection, severity of rejection, subclinical rejection and HLA mismatch. Alloimmune independent factors such as delayed graft function, donor age, Cytomegalovirus infection, donor/recipient co-morbidity and of course calcineurin inhibitor toxicity are important in the development of CAN. The pathogenesis of CAN is complex, multifactorial, and unfortunately incompletely understood. There are a number of pivotal steps in the initiation and propagation of the fibrosis seen in biopsy specimens from kidneys with CAN. Endothelial activation in response to one or more of the aforementioned risk factors stimulates leukocyte activation and recruitment. Recruited leukocytes subsequently infiltrate through the endothelium and induce key effector cells to secrete excessive and abnormal extracellular matrix (ECM). Enhanced deposition of ECM is a histological hallmark of CAN. This paper aims to present a concise yet accurate and up-to-date review of the literature concerning the aetiological factors and pathological processes which are present in the generation of CAN. PMID- 17138048 TI - Induction of endotoxin tolerance inhibits alloimmune responses. AB - It was recently reported that the induction of endotoxin tolerance (ET), which is defined as a reduced response to a lipopolysaccharide (LPS) challenge following the first LPS encounter, inhibits major histocompatibility complex (MHC) restricted antigen presentation. This raises the question whether alloimmune responses can be inhibited by inducing ET in transplant donors. C57BL/6 mice were treated with a low dose of LPS prior to a challenge with a high dose of LPS to induce ET. Hearts from endotoxin-tolerized C57BL/6 mice were transplanted to BALB/c mice. The survival of the endotoxin-tolerized heart allografts was significantly prolonged. By using irradiated splenocytes from C57BL/6 mice and allogeneic splenocytes from BALB/c mice, a mixed lymphocyte reaction (MLR) assay was performed. The MLR assay used CFSE, and revealed that the splenocytes from the endotoxin-tolerized mice failed to induce the proliferation of allogeneic CD4(+) and CD8(+) T cells. Cytokine analyses of the supernatant of the MLR culture using endotoxin-tolerized stimulators revealed a distinct shift in the Th 1/Th 2 balance toward the Th 2-type response. The induction of ET increased the proportion of myeloid-related dendritic cells (DCs) expressing molecules necessary for antigen presentation, which favor the development of a Th 2 response; however, it reduced the proportion of lymphoid-related DCs expressing those molecules, which favor the development of the Th 1 response. Although the relevance of these findings with regard to the prolonged survival of the endotoxin-tolerized heart allografts remains to be elucidated, this is the first study to demonstrate that the induction of ET in donor animals inhibits alloimmune responses. PMID- 17138049 TI - Effects of ex vivo activated immune cells on syngeneic and semi-allogeneic bone marrow transplantation in mice. AB - We have previously shown that a novel immunotherapy using ex vivo activated immune cells is capable of promoting survival and hematopoietic recovery in mice after combined chemotherapy and radiotherapy. In this study, we investigated whether the immunotherapy with ex vivo activated immune cells had the same beneficial effects after syngeneic and semiallogeneic bone marrow transplantation (BMT) in BALB/c mice subjected to a lethal dose of total body irradiation (TBI). Immune cells were cultured in vitro with a combination of cytokines and a calcium ionophore for 2 days and subsequently injected to mice daily for 4 days starting 1 day after BMT. The immunotherapy enhanced survival and multilineage peripheral blood recovery in BMT mice with limited numbers of transplanted bone marrow cells when a low dose of ex vivo activated immune cells were used. However, the beneficial effects were completely lost when a higher dose of the same therapeutic immune cells were tested, and instead the immunotherapy significantly exacerbated complications associated with the lethal radiation and BMT. This detrimental effect appeared to be the result of strong in vivo nonspecific immune responses induced by either activated therapeutic immune cells or interaction between therapeutic immune cells and MHC-mismatched bone marrow cells transplanted or both. Our data suggest that the immunotherapy with appropriately selected dosages may be beneficial to BMT but vigorous in vivo immune responses soon after BMT may exacerbate post-transplant complications. PMID- 17138050 TI - Circulating adhesion molecules during kidney allograft reperfusion. AB - Adhesion molecule expression is an important event during early transplant failure. The aim of the present study was to examine the release of adhesion molecules during the first minutes of kidney allograft reperfusion in relation to delayed graft function and acute graft rejection. We enrolled 49 renal transplant recipients, including 13 cases of delayed graft function (DGF) and 11 cases of acute graft rejection (AR). Plasma concentrations of E-selectin, VCAM-1 and ICAM 1 after 3 min of reperfusion were significantly higher than in the iliac vein before reperfusion. There was no statistically significant difference between patients with and without DGF as regards E-selectin, VCAM-1 and ICAM-1 concentrations in the iliac vein before and in the renal vein after 3 min of reperfusion. Concentrations of adhesion molecules in the iliac vein before reperfusion and in the renal vein after 3 min of reperfusion did not differ significantly between patients with and without AR except for ICAM-1 iliac vein concentration which was significantly increased in AR patients. Plasma levels of E-selectin, ICAM-1 and VCAM-1 were increased after kidney allograft reperfusion. Moreover, elevated serum levels of ICAM-1 before transplantation correlated with subsequent acute kidney allograft rejection. The results suggest that elevated ICAM-1 levels may be implicated in acute graft rejection. PMID- 17138051 TI - Glucose tolerance normalization following transplantation of pig pancreatic primordia into non-immunosuppressed diabetic ZDF rats. AB - Pancreas or pancreatic islet transplantation in humans is limited by organ availability, and success of the latter is negatively impacted upon by tissue loss post-transplantation and limited potential for expansion of beta cells. A way to overcome the supply and expansion problems is to xenotransplant embryonic tissue. Previously, we have shown that beta cells originating from embryonic day (E) 28 (E28) pig pancreatic primordia transplanted into the mesentery of streptozotocin-diabetic (type 1) Lewis rats engraft without the need for host immune-suppression and normalize glucose tolerance. Here we show long-term engraftment of pig beta cells within liver, pancreas and mesenteric lymph nodes post-transplantation of E28 pig pancreatic primordia into diabetic ZDF rats, a model for type 2 diabetes. Porcine insulin is present in circulation after an oral glucose load. Glucose tolerance is normalized in transplanted ZDF hosts and insulin sensitivity restored in formerly diabetic ZDF males. Release of porcine insulin in vitro from tissue originating in transplanted rats occurs within 1 min of glucose stimulation characteristic of first-phase secretion from beta cells. Of potential importance for application of this transplantation technology to treatment of type 2 diabetes in humans and confirmatory of our previous findings in Lewis rats, no host immunosuppression is required for engraftment of E28 pig pancreatic primordia. PMID- 17138052 TI - Knockdown of mouse VCAM-1 by vector-based siRNA. AB - Graft rejection is critically dependent on the recruitment of leukocytes via adhesion molecules on the endothelium, and inhibition of these interactions can prolong graft survival. We have therefore developed an approach using siRNA to inhibit the expression of VCAM-1 in endothelial cells. We transfected siRNA constructs into murine corneal and vascular endothelium and looked at expression of VCAM-1 and other surface molecules by flow cytometry. Adhesion assays (both static and under flow) were used to determine the effect of VCAM-1 inhibition. The activation of cellular stress responses was assessed by RT-PCR. Constructs encoding siRNA can block expression of VCAM-1 in both corneal and vascular endothelial cells (in the latter case after cytokine stimulation). Inhibition of VCAM-1 expression reduced the ability of T cells to adhere to endothelium. However, there were non-specific effects of siRNA expression, including upregulation of (Programmed Death Ligand 1) PDL1 and decreased cell growth. Analysis of stress pathways showed that the endothelial cells transfected with siRNA had upregulated molecules associated with cell stress. While these data are supportive of a potential therapeutic role for siRNA constructs in blocking the expression of adhesion molecules, they also highlight potential non-specific effects of siRNA that must be carefully considered in any application of this technology. PMID- 17138053 TI - Risk of adverse post-transplant events after kidney allograft transplantation as predicted by CTLA-4 +49 and TNF-alpha -308 single nucleotide polymorphisms: a preliminary study. AB - Genetic differences between donor and recipient HLA haplotypes are of major importance for transplant rejection. Other genetic variations occurring in genes encoding cytokines and costimulatory molecules also appear to exert an influence on the manner the host immune system recognizes the allograft. The aims of this work were: 1) to study selected single nucleotide polymorphisms (SNPs) at the loci encoding the T-cell regulatory molecule CTLA-4 (CD152), and the cytokines interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL 10, and transforming growth factor (TGF)-beta1 in a sample of healthy volunteers and a group of kidney-transplanted patients; and 2) to investigate whether an association exists between any of the SNPs studied and acute or chronic rejection, or non-responsiveness to steroid treatment during episodes of acute rejection (AR) after kidney allograft transplantation. When healthy volunteers were compared with transplanted patients, no significant differences were found in the distribution of genetic frequencies for any of the SNPs analyzed. However, in transplanted patients who received a kidney from a living related donor (KdTxL), a statistically significant association was found between carrying the CTLA-4 +49 A/A genotype and protection from experiencing acute rejection. No such association was found in the group of transplanted patients who received a kidney from a cadaveric non-related donor (KdTxCad). In both, KdTxL and KdTxCad patients, responsiveness to steroid treatment during acute rejection was also in association with the CTLA-4 (+49A/G) SNP. The CTLA-4 +49G allele was found at a very low frequency among steroid-resistant compared with steroid-sensitive patients. Finally, a statistically significant association was found between the presence of the TNF-alpha -308A allele and protection to suffer from chronic rejection. The genetic differences found may serve as risk predictors of adverse post-transplant events. PMID- 17138054 TI - Allogeneic activation is attenuated in a model of mouse lung perfused with magnesium-deficient blood. AB - Hypomagnesemia, which is frequently observed in patients treated with calcineurin inhibitors to prevent rejection after allogeneic transplantation, has been associated with a faster rate of decline in allograft function. The effect of hypomagnesemia on lung allograft has not been reported yet. In our model of isolated mouse lung, we have evaluated the early effects of allogeneic lung perfusion with blood from magnesium (Mg)-deficient mice for 3 h on lung activation and remodelling, compared to isogeneic perfusion. Hypomagnesemia (0.21+/-0.07 mmol Mg(2+)/l) was observed in blood from Mg-deficient mice, but no inflammatory pattern. The mRNA level of the intercellular adhesion molecule (ICAM)-1, but neither of the vascular cell adhesion molecule (VCAM)-1, nor of the cytokines tumor necrosis factor (TNF)alpha and interleukin (IL)-2, was enhanced (p<0.05). Although caspase-3 mRNA was transiently enhanced, no apoptotic cells were evidenced in lung tissues even after 3 h. Using cDNA array, we found that the genes encoding RANKL, RANK, TNFR2, NFATX, IL-1R2, IL-6R gp130, SOCS3, PDGFRB, P63, CSF3R, CXCL1, CXCL5, CX3CL1, CSF1, which are involved in inflammation and apoptosis regulation, were markedly up-regulated in allogeneic conditions. Our results support a limited allogeneic activation and an early stage of the inflammatory process in lung, at the time of inflammatory cell recruitment without lung tissue remodelling, as a result of hypomagnesemia. These findings suggest that cyclosporine-related hypomagnesemia, observed in most of the transplanted patients, does not constitute an additional risk for lung allograft outcome. PMID- 17138055 TI - T-cell inhibition does not aggravate bacterial translocation from rat small bowel. AB - BACKGROUND: T-cell mediated immunity has been proposed to have an important function in the defence against translocating microbes from the gastrointestinal tract. After small bowel transplantation massive T-cell immunosuppression is necessary to avoid rejection. As a consequence, infections with intestinal bacteria are the main contributors to mortality in this setting. This could further imply that T cells are important in limiting bacterial translocation. In a model for bacterial translocation from small bowel in the rat we examined the outcome of T-cell inactivation. METHODS: The studies were performed in a model of bacterial translocation from a Thiry-Vella loop of small bowel in the rat. The animals were treated with an anti-alpha/beta T-cell receptor monoclonal antibody (R73). Inhibition of T-cell activation was also made using the immunosuppressive drug cyclosporin A. All animals were sacrificed on day 3 postoperatively and translocation to the mesenteric lymph nodes, liver, spleen, lung and blood was evaluated. RESULTS: Treatment with R73 resulted in an almost complete labelling of T cells but did not result in any increased bacterial translocation compared to animals treated with saline. Neither did immunosuppression with cyclosporin A. CONCLUSIONS: In the model of bacterial translocation from a defunctionalised loop of small bowel the inhibition of T cells does not increase bacterial translocation to mesenteric lymph nodes or promote the systemic spread of the translocating bacteria. This indicates that T cells do not have any important protective function against translocating microbes from defunctionalised small bowel. PMID- 17138056 TI - Lymphocyte propagation from biopsies of kidney allografts. AB - Morphological evaluation of transplant biopsies, usually using the Banff classification, is the most important tool to diagnose rejection after kidney transplantation. However, morphological analysis only scores the amount and localisation of infiltrating cells, and studies show that up to 30% of grafts with a stable function display infiltration of lymphocytes consistent with acute cellular rejection. Methods to study the functional properties of the infiltrating lymphocytes are therefore needed. We applied a tissue culture system on biopsies from transplanted human kidneys, allowing infiltrating cells to propagate out from the tissue. Cells were then counted and subtyped by flow cytometry. The results were correlated to morphology. In total, 92 biopsies from 69 patients were analysed. For 14 patients, serial biopsies were available. In grafts with cellular or combined cellular and vascular rejection, the number of ex vivo propagated mononuclear cells was higher than from non-rejecting grafts. A similar pattern was seen for CD3(+) T cells as well as for T cells expressing CD25 or MHC class II antigens. However, the proportion of CD25(+) or MHC class II(+) T lymphocytes was similar in all groups (no rejection, vascular rejection, borderline changes, cellular rejection, combined cellular and vascular rejection). In all groups the number of CD4(+) cells was higher than the number of CD8(+) cells. The results confirm previous experimental studies showing that graft-infiltrating cells are possible to culture in vitro and that lymphocyte propagation correlates to acute cellular rejection. Tissue culturing is easy to perform and evaluate and can be used to determine and analyse the cellular immune response to allografts and may thus be used as a complement to morphological analyses. PMID- 17138057 TI - The nonopsonic allogeneic cell phagocytosis of macrophages detected by flow cytometry and two photon fluorescence microscope. AB - Phagocytosis, one of the apparent functions for macrophages, represents an early and crucial event in triggering host defenses against invading pathogens as well as allo- or xenogeneic rejection. Now, some methods have been used in detecting the opsonic phagocytosis of macrophages in xenogeneic settings. Efficient nonopsonic phagocytosis analysis method has not been established yet. In the present studies, allogeneic lymphocytes pre-labeled with 5-(and-6) carboxyfluorescein diacetate succinimidyl ester (CFSE) or derived from green fluorescent protein transgeneic B6 mice (GFP-B6 mice) were co-incubated with primary murine peritoneal macrophages (PEMs) for 1-2 h or were injected into murine peritoneal cavity for 30 to 240 min. Assays by flow cytometry (FCM) and two photon laser scanning microscope (TPM) showed an efficient uptake of both allogeneic lymphocytes and xenogeneic chicken red blood cells. The continuing process of nonopsonic phagocytosis of allogeneic lymphocytes by PEMs was recorded by TPM. Furthermore, the phenotype differences of PEMs with or without phagocytosis of allogeneic cells were determined by three-color FCMs. Significantly upregulated expressions of CD11b, CD44, TLR2 and TLR4 on PEMs were observed as early as 6 h after phagocytosis of allogeneic cells. Our present data indicated that the FCM and TPM combined method is a practical approach to detect macrophage nonopsonic phagocytosis of allogeneic lymphocytes and to identify the phenotype alteration of macrophages after phagocytosis. PMID- 17138058 TI - sCD30, interleukin-1beta-converting enzyme and anti-Annexin V autoantibodies concentrations in heart transplant recipients. AB - sCD30 and ICE/caspase-1 as apoptosis-regulating factors are suspected to be involved in the survival rate of immunocompetent cells during immunosuppression after allotransplantation. Serum CD30 and ICE/caspase-1 concentrations were estimated and associated with unspecific serum apoptosis marker--anti-Annexin V antibodies and myocardial biopsies results. MATERIALS AND METHODS: 28 clinically stabile patients--heart transplant recipients at least 3 months after cardiac transplantation performed due to heart failure caused by ischaemic and/or congestive cardiomyopathy or/and primary valvular heart disease (26 men and 2 women, mean age=36.8 years, S.D.=7.6) with normal heart function assessed by use of ultrasound scan--were involved in the trial. The patients were divided and analyzed in two ways: first according to the results of elective endomyocardial biopsies and second to main immunosuppressive agent used. The enzyme immunoassay (CD30, Dako; interleukin-1beta-converting enzyme (ICE)/Caspase-1 ELISA and anti Annexin V BENDER MedSystem) for soluble CD30, caspase-1 and anti-Annexin V autoantibodies serum levels was used. RESULTS: sCD30 and caspase-1 concentrations were non-significantly up-regulated in all analysed groups--with or without rejection signs or immunosuppressed with cyclosporine or especially tacrolimus. In contrast anti-Annexin V autoantibodies concentration was non-significantly down-regulated also in all studied groups. Moreover in the group with signs of transplant rejection, strong negative correlation between anti-Annexin antibodies and rejection grade was observed (-0.65, p<0.05). Biopsy results were comparable in groups treated with tacrolimus and cyclosporine A. CONCLUSIONS: The increasing tendency of sCD30 and caspase-1 as well as the decrease in anti-Annexin V autoantibodies concentrations in heart recipients could be the result of post transplant apoptosis disturbances. This tendency seems to be inhibited in a greater degree by tacrolimus than by cyclosporine. Anti-Annexin V autoantibodies might be considered as negative rejection markers due to their strong negative correlation with the rejection grade. PMID- 17138059 TI - Modified ELISPOT technique--highly significant inverse correlation of post-Tx donor-reactive IFNgamma-producing cell frequencies with 6 and 12 months graft function in kidney transplant recipients. AB - BACKGROUND: Upcoming trials for immunosuppression minimization and tolerance induction require the development of reliable in vitro assays for monitoring cellular alloimmunity in transplant patients. The IFN-gamma ELISPOT assay represents a promising tool for monitoring alloreactive memory/effector T cells. As T lymphopenia is a common finding during the early post-transplant (post-Tx) period, the IFN-gamma ELISPOT technique was here modified by using ELISPOT responder cells with enhanced percentage and standardized number of 200,000 T cells per well. METHODS: Peripheral blood mononuclear cells (PBMNC) of kidney transplant recipients were depleted of CD14+ and CD15+ cells to increase the percentage of T cells from average 47.8% to 71.5% before transplantation (pre-Tx) and from 39.7% to 74.9% post-Tx. The assay was tested in a population of 23 de novo renal transplant patients for clinical relevance. Before and at 2-3 times during the first 6 months post-Tx, IFN-gamma-producing donor-reactive as well as recall antigen-reactive cell frequencies (Candida, tuberculin, tetanus) were determined and correlated with outcome. RESULTS: Pre-Tx donor-reactive ELISPOT frequencies were enhanced in patients with acute rejection compared to non rejectors. Moreover, mean post-Tx donor-reactive ELISPOT frequencies showed a highly significant inverse correlation with renal function at 6 and 12 months. In contrast, recall antigen-reactive ELISPOT frequencies did not correlate with outcome. CONCLUSION: Our results suggest that the modified donor-reactive ELISPOT approach might provide a useful surrogate marker for renal transplant outcome with possible utility especially in T-lymphopenic patients. PMID- 17138060 TI - TIRC7 is induced in rejected human kidneys and anti-TIRC7 mAb with FK506 prolongs survival of kidney allografts in rats. AB - TIRC7 delivers essential signals during immune activation as antibodies targeting TIRC7 inhibit lymphocyte proliferation and Th1 cytokine expression in vitro and prolonged kidney and heart allograft survival in vivo. Immunohistochemical analysis of biopsy specimens from human renal allografts undergoing rejection despite treatment with Calcineurin inhibitors (CI) showed elevated TIRC7 expression. Accordingly, with a view to clinical application, we evaluated the therapeutic effect of a chimerized anti-TIRC7 mAb in combination with Tacrolimus (FK506) using a rat kidney transplantation model (DA to Lewis). The combination of sub-therapeutic doses of both compounds significantly (p<0.05) prolonged the median graft survival to 19.5 days compared to monotherapy with FK506 (median survival, 7d) or mAb against TIRC7 (7d). These results suggest a potential synergism of anti-TIRC7 mAb and FK506 action, which could be developed into a novel combination therapy in the clinic by lowering side effects of present CI treatment. Moreover, the identification of TIRC7 in graft infiltrating lymphocytes might serve as a diagnostic marker to detect allograft rejection. PMID- 17138061 TI - Synthetic blood group antigens for anti-A removal device and their interaction with monoclonal anti-A IgM. AB - Removal of blood group antibodies against the donor organ prior to ABO incompatible transplantation can prevent episodes of hyperacute rejection. We are developing a specific antibody filter (SAF) device consisting of immobilized synthetic Atrisaccharide antigens conjugated to polyacrylamide (Atri-PAA) to selectively remove anti-A antibodies directly from whole blood. In this study, we evaluated eight anti-A IgM monoclonal antibodies (mAbs) using Enzyme-Linked Immunosorbent Assay (ELISA) to determine their specificity for binding to Atri PAA. Five of the eight mAbs met our criteria for specificity by binding to Atri PAA with at least five times greater affinity compared to the negative controls. These selected mAbs will be studied for their binding characteristics to Atri-PAA which will aid in the development of the SAF. The study of kinetics of antibody removal and quantification of antibody removal will be used in our mathematical model to maximize the antibody removal rate and binding capacity of the SAF. PMID- 17138062 TI - Late onset of severe graft-versus-host disease following liver transplantation. AB - Graft versus host disease (GVHD) is an uncommon but lethal complication following liver transplantation that results from the engraftment of T lymphocytes associated with the liver graft. It usually occurs 2 to 6 weeks after the procedure. We herein report a case of late onset of severe GVHD 4 months after cadaveric liver transplantation for hepatocellular carcinoma in a 54-year-old woman, which was characterized by refractory diarrhea and abdominal pain. Moreover we discuss risk factors of GVHD including the recipient age and cytomegalovirus (CMV) infection. PMID- 17138063 TI - Alemtuzumab (Campath-1H) induction therapy and dendritic cells: Impact on peripheral dendritic cell repertoire in renal allograft recipients. AB - Dendritic cells (DC) are the most potent antigen-presenting cells (APC) and are pivotal for initiating allograft immunity. Recently, particular DC subsets have been implicated also in allogeneic T cell hyporesponsiveness. Alemtuzumab (anti CD52, Campath-1H) is a novel T cell depleting antibody that is currently under investigation for the use in allogeneic organ transplantation. While recent studies demonstrated a conspicuous effect of alemtuzumab on peripheral DC in clinical graft-versus-host disease, its efficiency in patients receiving allogeneic organ transplants is still undefined. In the present study we assessed the peripheral DC repertoire in kidney transplant recipients after either alemtuzumab induction therapy followed by FK506 monotherapy or after conventional immunosuppression (FK506, mycophenolate mofetil and steroids) without any induction agent. Induction with alemtuzumab caused a strong and sustained reduction of the total number of peripheral DC and a significant shift from myeloid to plasmacytoid DC subsets (mDC/pDC ratio) as early as 1 month post transplantation. These data show that alemtuzumab induction targets the peripheral DC repertoire, which might add another mechanism allowing immunosuppressive drug minimization. Further studies are warranted to further elucidate the functional significance of these finding in the setting of allogeneic organ transplantation. PMID- 17138064 TI - The impact of donor cytokine gene polymorphisms on the incidence of cytomegalovirus infection after kidney transplantation. AB - Cytomegalovirus (CMV) seronegative recipients of kidneys from CMV seropositive donors are at a high risk of CMV infection after transplantation since viruses in the allograft may reactivate in patients without prior immunity. We hypothesized that the genetic background of the graft has an influence on the incidence of infection. Effects of IL10, IL6 and IFNG gene polymorphisms, known to affect CMV infectivity, were investigated in 71 CMV seronegative recipients of grafts from CMV seropositive cadaver donors. Donor IL10(-1082 AA) genotype reduced the incidence of CMV infection (p=0.031) and CMV episodes in these patients tended to occur later (AA: median 83 days, AG/GG: median 45 days, p=0.072). In multivariate analysis, other explaining factors than the donor IL10(-1082 AA) genotype alone did not improve Cox hazard model (HR=0.3, 95% CI=0.09-0.96, p=0.043). Recipient polymorphisms did not reduce the incidence of CMV infection. We conclude that donor IL10 gene polymorphisms may influence the likelihood of CMV infection in the high risk patients investigated. PMID- 17138065 TI - Differential risks of subsequent vascular events for transient ischaemic attack and minor ischaemic stroke. AB - Using a prospective hospital-based registry, 146 patients with transient ischaemic attack (TIA) were compared with 376 patients with minor first-ever ischaemic stroke with respect to the 3-month risk of subsequent vascular events, in order to clarify the distinctions between the disease entities. All patients were enrolled within 48 h of onset. The risk factor distribution for the two groups was comparable, except that the TIA patients had more previous TIAs. Large artery atherosclerosis (34%) and small vessel occlusion (32%) were the main aetiologies in the TIA group, whereas small vessel occlusion (49%) was the major cause in the stroke group. The 3-month risk of combined endpoints of stroke, myocardial infarction, and vascular death for TIA patients was higher than that for the minor stroke group (15.1% vs. 3.2%; hazard ratio 4.6, 95% confidence interval 2.3-9.3 in multivariate analysis). Large artery atherosclerosis and male sex were the other significant predictors. TIA may demand more urgent management than minor stroke. The fact that aetiology is a predictor, highlights the need for rapid diagnostic tests to establish pathogenesis. PMID- 17138066 TI - Conduction slowing in painful versus painless diabetic neuropathy. AB - Motor conduction slowing in diabetic distal symmetrical polyneuropathy (DSP) generally exceeds that in distal axonal polyneuropathy. Additional mechanisms secondary to axonal injury may contribute to motor conduction slowing; however, its clinical and pathophysiological significance has rarely been discussed. The purpose of this study was to evaluate the clinical and pathophysiological significance of conduction slowing in DSP. We analyzed motor conduction in 50 patients with symptomatic painful DSP and 25 patients with asymptomatic painless DSP. Motor conduction data from 40 patients with amyotrophic lateral sclerosis (ALS) were used as controls for pure axonal conduction slowing. Compound muscle action potential amplitude (CMAP), distal motor latency (DL), and conduction velocity (CV) values were converted to a percentage of the upper or lower limits of normal and then presented in square root transformation (SQRT) form. Plots of SQRT-DL and SQRT-CV against SQRT-CMAP were analyzed. Regression analysis showed that DL and CV are amplitude-dependent in both painless DSP and ALS. Changes of DL and CV in painful DSP were not amplitude-dependent except for DL in the lower extremities. Our data support the hypothesis that the mechanism of slowing is similar in both painless DSP and ALS, and that it results from the loss of large, fast-conducting fibres. Lack of amplitude-dependency in conduction slowing in painful DSP may reflect combined axonal and demyelinating changes. PMID- 17138067 TI - P300 and treatment effect of modafinil on fatigue in multiple sclerosis. AB - To evaluate the value of visual and auditory P300 for predicting the response of multiple sclerosis-related fatigue to modafinil treatment, 33 patients were treated with 100 mg modafinil once daily for 4 weeks, following a 4-week baseline phase and an optional 8-week extension phase. The main clinical outcome parameter was a decrease in the fatigue visual analogue score (VAS) before and after 4 weeks of treatment. Patients with shorter auditory P300 latency at baseline were more likely to benefit from modafinil treatment. Auditory P300 latency predicted treatment response with a specificity of 76% and a sensitivity of 75% at a cut off latency of 350 ms. Visual P300 latency could not be used to predict treatment response. Baseline auditory P300 latency predicted treatment response, whereas visual P300 latency did not. Clinical improvement did not correlate with changes in either visual or auditory P300. PMID- 17138068 TI - Role of the activated extracellular signal-regulated kinase pathway on histological and behavioral outcome after traumatic brain injury in rats. AB - The extracellular signal-regulated kinase (ERK) pathway, which modulates the activity of many transcriptional factors leading to the proliferation of various cells, is activated in lesions in regions of selective vulnerability after traumatic brain injury (TBI). In the present study, using the ERK inhibitor U0126, we investigated the role of the ERK pathway in histopathological and behavioral outcomes after TBI. Adult male Sprague-Dawley rats, weighing 300-400 g were subjected to lateral fluid percussion brain injury. The ERK inhibitor U0126 was injected intravenously before injury at 100, 200 and 400 microg/kg. The severity of CA3 neuronal damage was evaluated by the number of surviving CA3 neurons 7 days after injury. The contusional lesion volume 72 h after injury was analysed using a computer-assisted analysis system. Three different motor skill tasks were measured on days 1-5, 7, 14 and 21 after injury. Pretreatment with U0126 significantly reduced both CA3 neuronal damage and contusional lesion volume after injury. In addition, administration of U0126 ameliorated motor function recovery on days 3, 4 and 5 after injury. Therefore, inhibition of ERK phosphorylation could be a potentially effective therapeutic target after TBI. PMID- 17138069 TI - Spinal instrumentation for interfacet locking injuries of the subaxial cervical spine. AB - A retrospective analysis of our surgical management of traumatic interfacet locking was performed. Eleven interfacet locking injuries were surgically treated. An anterior procedure was performed in five patients, posterior fixation in three and a combined procedure in three. Five facet locks were reduced by preoperative skull traction. After general anesthesia at surgery, another two cases were corrected manually. Surgical fixation using spinal instrumentation was performed. One patient treated with posterior fixation required an additional anterior procedure because of a delayed disc herniation. Spinal instrumentation avoided a halo vest. The anterior approach may be selected in patients who are reduced manually, while a combined procedure should be performed in patients with irreducible facet dislocation with disc herniation. Delayed symptomatic disc herniation may occur when only posterior fixation is performed. PMID- 17138070 TI - Malignant transformation of high-grade astrocytoma associated with neurocysticercosis in a patient with Turcot syndrome. AB - A 45-year-old woman with anaplastic astrocytoma was clinically diagnosed with Turcot syndrome, and subsequently developed simultaneous neurocysticercosis and malignant transformation to glioblastoma. The parasitic cysts and glioblastoma were microsurgically removed. Histological examination of surgical specimens revealed neurocysticercosis between the normal brain tissue and glioblastoma. The clinical course and histological findings suggest that the parasitic infection and/or genetic changes contributed to the malignant transformation of the astrocytic tumour. PMID- 17138071 TI - Migration of a bullet in the spinal canal. AB - Migration of a bullet within the spinal canal after gunshot injury is rare. We report here the case of a penetrating gunshot injury of the lumbar spine at L3 with migration of the bullet within the spinal canal S2. The patient had marked paraparesis (proximal 1/5, distal 0/5 muscle strength) and anaesthesia at L3 and below, and had a hypocompliant, hyper-reflexive bladder with decreased capacity, and absent anal tonus. We removed osseous fragments in the canal with an L3 laminectomy and extracted the bullet by S2 laminectomy. After surgery, we observed an improvement in paraparesis, an increase in bladder capacity and urinary compliance, and improvement in anal tonus. The appropriate course of action in this type of injury remains unclear, because the number of cases described in the literature is not sufficient to provide a basis on which to make a definitive therapeutic decision. We herein review the literature describing cases in which a bullet in the spinal canal has migrated; we describe the treatment used and the outcomes in these cases. PMID- 17138072 TI - Radiological findings for arterial dissection of the anterior cerebral artery. AB - We treated a patient with anterior cerebral artery (ACA) dissection that caused an ischaemic stroke, and investigated serial changes over time by using three different radiological methods. The conventional angiography findings for ACA dissection corresponded to those of computed tomography (CT) angiography, but not those of magnetic resonance angiography for each phase. We presume that the results were based on the velocity of the blood flow in the pseudolumen of the dissected artery, and we believe that CT angiography is a useful and less invasive diagnostic tool for intracranial arterial dissection. PMID- 17138073 TI - Intracerebral haemorrhage during surgery for chronic subdural haematoma. AB - Although intracerebral haemorrhage occurring after surgery for chronic subdural haematoma has been reported as a rare complication, intracerebral haemorrhage occurring during surgery has not been described previously. A case of bilateral chronic subdural haematomas, with a small motor area haemorrhage occurring intra operatively, is described here. PMID- 17138074 TI - Cerebellar abscess and syringomyelia due to isoniazid-resistant Mycobacterium tuberculosis. AB - A 19-year-old immunocompetent man was admitted to hospital with diplopia, nausea, vomiting and change in mental status. The patient had a history of tuberculous meningitis that was diagnosed at another hospital 6 months before the present admission, and at that time anti-tuberculosis treatment was initiated using a first-line drug combination. A computed tomography (CT) scan of the brain revealed non-communicating hydrocephalus. A ventriculo-peritoneal shunt was inserted surgically. Two months later, the patient was hospitalized again for fever, dysphagia and left hemiparesis. At that time, his cranial CT findings were within normal limits; however, magnetic resonance imaging (MRI) revealed an irregular multilocular peripheral contrast-enhancing lesion in the posterior fossa. The abscess was surgically drained. The presence of acid-fast bacilli in the abscess material was demonstrated by Ziehl-Neelsen staining. Mycobacterium tuberculosis grew on Lowenstein-Jensen culture medium, and the strain was found to be resistant to isoniazid. One month after the operation, the patient became quadriparetic. Cervical MRI revealed a cervico-thoracic syringomyelitic cavity, after which a syringoperitoneal shunt was placed. Treatment with four drugs was continued for 10 months, and then treatment with three drugs for a total period of 18 months. The patient recovered, with residual quadriparesis. Even though very rare, isoniazid-resistant M. tuberculosis may be the causative agent of progressive tuberculosis. PMID- 17138075 TI - Identification of a novel mutation in a Korean patient with oculopharyngeal muscular dystrophy. AB - Oculopharyngeal muscular dystrophy (OPMD) is a late-onset muscle disorder characterized by progressive dysphagia and bilateral ptosis. Mutations in the polyadenylate binding protein nuclear 1 (PABPN1) gene have been found to cause OPMD. The typical mutation is a stable trinucleotide repeat expansion in the first exon of the PABPN1 gene, in which (GCG)(6) is the normal repeat length. We investigated a Korean patient with OPMD and identified a novel mutation: a heterozygous insertion of a 9-bp sequence [(GCG)(GCA)(GCA); c.27_28insGCGGCAGCA] instead of the (GCG) repeat expansion, resulting in an in-frame insertion of three alanines (p.A10insAAA). To the best of our knowledge, this is the first report of a genetically confirmed case of OPMD in Korea. PMID- 17138076 TI - The time is ripe. PMID- 17138077 TI - Impact of intraoperative transesophageal echocardiography during noncardiac surgery. AB - OBJECTIVE: The purpose of this study was to test if parameters measured by intraoperative transesophageal echocardiography (TEE) could be useful to evaluate the hemodynamic status of high-risk cardiovascular patients and if this information was sufficient to make changes in intraoperative management. DESIGN: Prospective clinical study. SETTING: Single-university hospital. PARTICIPANTS: Ninety-eight patients undergoing noncardiac surgery. INTERVENTIONS: Every patient was assessed with a baseline examination of 2-dimensional, color, pulsed, and continuous Doppler images. Intraoperative changes in any of the evaluated and measured parameters led to a specific change according to the protocol. MEASUREMENTS AND MAIN RESULTS: After continuous monitoring with TEE during surgery, all patients were assigned to 1 of the following groups: (1) TEE was of no use, (2) TEE-directed intraoperative management changes, (3) intraoperative TEE-directed changes in postoperative management, and (4) TEE successfully used as a substitute for pulmonary artery catheter monitoring. Two patients (2%) were assigned to group 1, 47 (48%) patients to group 2, 25 (25%) patients to group 3, and 24 (24%) patients to group 4. The most frequent modifications in intraoperative management were changes in drug therapy and fluid administration. Postoperative management changes were mostly made because of new diagnosis (14%) and new left ventricular wall motion abnormalities (9%). CONCLUSION: These results strongly suggest that objective measurements made by intraoperative TEE are effective in unveiling relevant clinical findings and useful information in high-risk patients undergoing noncardiac surgery. PMID- 17138078 TI - The impact of spontaneous echocardiographic contrast in patients with left atrial enlargement undergoing cardiac valvular surgery. AB - OBJECTIVE: The aim of this study was to evaluate the impact of either left atrial or aortic spontaneous echocardiographic contrast (SEC), as identified on intraoperative transesophageal echocardiography, on short-term morbidity and mortality in patients with left atrial enlargement undergoing cardiac valvular surgery. DESIGN: Retrospective and observational. SETTING: Single-center, university teaching hospital. PARTICIPANTS: The authors identified 197 patients (105 males and 92 females; mean age, 68 +/- 14 years) with left atrial enlargement who underwent surgical intervention for valvular heart disease from January 1, 2004 to January 1, 2005. MAIN RESULTS: Of the total population, 40 patients (20.3%) showed left atrial SEC, and 10 patients (5.1%) showed aortic SEC. On multivariate analysis, increasing left atrial size and the absence of mitral regurgitation were independent predictors for the presence of left atrial SEC. On multivariate analysis, the presence of atrial fibrillation and a dilated descending aorta were predictive of aortic SEC. Although the identification of left atrial SEC was an echocardiographic marker of an increased risk for thromboembolic events postoperatively, this finding did not hold true for the presence of aortic SEC. CONCLUSIONS: Intraoperative identification of left atrial dilatation or aortic dilatation is predictive of SEC in the left atrium or descending aorta, respectively. The identification of left atrial SEC is an echocardiographic marker of an increased risk for thromboembolic events in this high-risk population. PMID- 17138079 TI - Implication of the anatomy of the pericardial reflection on positioning of central venous catheters. AB - OBJECTIVE: Central venous catheterization is associated with a significant incidence of complications (5%-20%). The incidence of perforation is approximately 0.25% to 0.4%. To prevent cardiac tamponade associated with a high risk of death, Food and Drug Administration guidelines state that the tip of a central venous catheter (CVC) should not be placed in, or allowed to migrate into, the heart. Therefore, in order to prevent cardiac tamponade, a catheter should be placed above the pericardial reflection. Thus, the intrapericardial length of the superior vena cava (SVC) was studied. Neither the pericardial reflection nor the exact entrance to the right atrium (RA) can be identified by chest x-ray. The goal of this study was to evaluate the variability of the intrapericardial section in relation to the SVC. DESIGN: Observational study. INTERVENTIONS: The absolute length of the SVC, the upper edge of the pericardial reflection on the SVC, and the lateral and the medial intrapericardial sections of the SVC were recorded and statistically analyzed. SETTING: Medical school: dissecting room at the Department of Anatomy. STUDY POPULATION: Eighteen formalin preserved adult cadavers. MEASUREMENTS AND MAIN RESULTS: The median lengths measured were as follows: total SVC, 61 mm; intrapericardial section of the medial SVC, 32.5 mm; and lateral SVC, 20.5 mm. The intrapericardial section was related to the total length of the SVC on both sides (Spearman rank order, p < 0.05). The median difference of the SVC covered with pericardium between the lateral and medial side was 11 mm (range, 5-21). In 15 of 18 cadavers, the pericardial reflection ran within the medial third of the SVC. The lower third of the SVC was regularly covered by the pericardium. The duplication of the pericardium crossed the SVC in the medial third at a diagonal to horizontal angle. CONCLUSIONS: Catheters ending below the pericardial reflection, hence positioned in the caudal third of the SVC, are likely to run along the long axis of the vein and the risk for perforation is minimized. Therefore, the authors recommend placing all catheters below the pericardial reflection. According to the present data, CVCs placed approximately 30 mm above the RA border, thus complying with the Food and Drug Administration guidelines, still may have their tips positioned below the pericardial reflection. In this position, pericardial tamponade still may occur. Perforation above the pericardial reflection will result in a hemo- or hydrothorax/mediastinum. A bedside method to determine the position of the CVC with respect to the pericardial reflection (eg, electrocardiographic guidance) should be used. PMID- 17138080 TI - Long-term outcome of patients with perioperative myocardial infarction as diagnosed by troponin I after routine surgical coronary artery revascularization. AB - OBJECTIVE: Diagnosis of perioperative myocardial infarction (P-MI) after coronary artery bypass graft (CABG) surgery traditionally relied on a combination of electrocardiographic and enzyme assay changes. Patients with Q-wave P-MIs who survive to hospital discharge have a poorer long-term prognosis. Troponin assays are more sensitive and specific for detecting minor P-MI, with an increased incidence of P-MI being reported. This study investigated if P-MI after CABG surgery, as defined by troponin-I isozyme (cTn-I), correlated with long-term outcome. DESIGN: A prospective, observational study. SETTING: A single institution, cardiothoracic specialty hospital. PARTICIPANTS: Seventy patients undergoing elective CABG surgery. INTERVENTIONS: Patients (n = 70) were stratified into low-risk and high-risk groups according to the absence (cTn-I <15 microg/L) or presence (cTn-I >or=15 microg/L) of P-MI after CABG surgery. Patients with (n = 24) and without (n = 46) P-MI were then followed for 3 years after CABG surgery to determine the impact of cTn-I-defined P-MI on long-term outcome. MEASUREMENTS AND MAIN RESULTS: Most patients felt that their quality of life and activity index had improved and that their symptoms of angina had lessened at 12-month follow-up. However, cardiovascular event-free survival was significantly less in patients with P-MI (p = 0.01) 3 years postoperatively. The incidence for cardiovascular events was 0.24 versus 0.65 (p = 0.049) in those patients without and with P-MI, respectively. The hazard ratio (2.9; 95% confidence interval, 1.3-9.4) for cardiovascular incidents was also significantly greater in patients with P-MI. More specifically, the incidence of arrhythmia was 2.4% versus 26.1% (p < 0.01), and the incidence of vascular events was 4.9% versus 26.1% (p = 0.02) in patients without and with P-MI, respectively. CONCLUSIONS: It was shown that P-MI as defined by cTn-I is associated with an increased long-term incidence of adverse cardiovascular events. An elevated peak cTn-I level (>or=15 microg/L) identified patients at increased risk but did not have a powerful positive predictive value for either cardiovascular (48%) or vascular (26%) complications. However, a peak cTn-I <15 microg/L was a negative predictor of adverse vascular outcome (95%). This may have implications for postoperative patient follow-up. PMID- 17138081 TI - Temperature management during off-pump coronary artery bypass graft surgery: a randomized clinical trial on the efficacy of a circulating water system versus a forced-air system. AB - OBJECTIVE: The aim of this study was to evaluate the performance of a new temperature management system specifically designed for cardiac surgery (Allon ThermoWrapping Thermoregulation System; MTRE Advanced Technologies Ltd, Or Akiva, Israel) using a circulating-water garment and to compare it with a conventional forced-air cover system in order to determine whether it could reduce the incidence of perioperative hypothermia during off-pump coronary artery bypass graft (OPCAB) surgery. DESIGN: Prospective, randomized. SETTING: University, tertiary care hospital. PARTICIPANTS: Thirty-one patients undergoing primary OPCAB surgery. INTERVENTIONS: Patients undergoing OPCAB surgery were randomized into the new thermoregulation system, Allon (study group, n = 15), and the standard forced-air system, Bair Hugger (Sterile Cardiac Access blanket Model 645; Augustine SA, Berne, Switzerland) (control group, n = 16). MEASUREMENTS AND MAIN RESULTS: Rectal temperature was recorded each 30 minutes during surgery and at intensive care unit arrival. Patients in the study group had higher temperatures than the control group at all time points, and the difference reached statistical significance after 2 hours of surgery. Moreover, fewer patients in the study group suffered perioperative hypothermia (defined as rectal temperature <36 degrees C) than the control group (2/15 patients (13.3%) in the study group v 13/16 (81.3%) in the control group [p = 0.0006]). No difference in other outcomes was noted. None of the patients died in the hospital. There were no adverse events reported. CONCLUSIONS: The circulating-water garment, Allon ThermoWrapping Thermoregulation System, maintained normothermia during OPCAB surgery better than forced-air systems, especially after the first 2 hours of surgery, and it was not associated with surgical field disturbance. PMID- 17138082 TI - Bolus administration of landiolol, a short-acting, selective beta1-blocker, to treat tachycardia during anesthesia: a dose-dependent study. AB - OBJECTIVES: The purpose of this study was to clarify the clinical efficacy of landiolol, a selective beta(1)-blocker, in patients developing tachycardia during anesthesia. DESIGN: A prospective, randomized, and placebo-controlled study. SETTING: A single university hospital. PARTICIPANTS: Patients undergoing resection of intracranial or maxillofacial tumors under general anesthesia. INTERVENTIONS: Intravenous bolus administration of landiolol. MEASUREMENTS AND MAIN RESULTS: Patients with heart rates exceeding 90 beats/min for more than 5 minutes received an intravenous bolus dose of landiolol. These patients who developed tachycardia were randomized into 4 groups receiving landiolol in the dose of 0, 0.1, 0.2, or 0.3 mg/kg. Heart rate and blood pressure were recorded before drug injection and 1, 3, 5, 7, 10, 15, 20, 25, and 30 minutes after the injection. Heart rate was significantly reduced with each dose of landiolol compared with the placebo group between 3 and 20 minutes after drug injection. The lowest heart rate was 93 +/- 4, 80 +/- 8, 79 +/- 8, and 77 +/- 6 beats/min (mean +/- standard deviation) in the landiolol 0, 0.1, 0.2, and 0.3 mg/kg groups, respectively. On the other hand, reduction of blood pressure was not as remarkable as that of heart rate. Landiolol, 0.1 and 0.2 mg/kg, did not significantly reduce systolic or diastolic blood pressure during the study period, although significant reduction of blood pressure was observed in the landiolol 0.3-mg/kg group between 5 and 10 minutes after drug injection. CONCLUSION: The results showed that bolus administration of landiolol was effective in the treatment of tachycardia during anesthesia in surgical patients. Landiolol was more efficacious in reducing heart rate than blood pressure. PMID- 17138083 TI - Increased risk associated with combined carotid endarterectomy and coronary artery bypass graft surgery: a propensity-matched comparison with isolated coronary artery bypass graft surgery. AB - OBJECTIVE: Risk associated with combined carotid endarterectomy and coronary artery bypass graft surgery (CEA/CABG) is controversial. The present study objective was to compare morbidity and mortality outcomes in well-matched patients who underwent combined CEA/CABG surgery with patients undergoing isolated CABG surgery with and without a history of a prior CEA. DESIGN: This investigation was designed as a retrospective case-controlled study using data from the Cardiothoracic Anesthesia Patient Registry in a single tertiary institution. The patient population consisted of 1,698 isolated CABG surgery patients with carotid artery stenosis >40%, 708 patients who underwent an isolated CABG surgery but had a history of a prior CEA, and 272 combined CEA/CABG surgery patients who underwent surgery from January 4, 1993, through June 30, 2003. Propensity modeling techniques were used to calculate a propensity score for each patient. Greedy matching resulted in 272 propensity-matched pairs of combined CEA/CABG and isolated CABG patients (primary analysis) and 241 propensity-matched pairs of combined CEA/CABG surgery and isolated CABG surgery with previous CEA patients (secondary analysis). A Fisher exact, chi-square, Wilcoxon rank sum, and Student t test were applied appropriately to compare the propensity-matched pairs. RESULTS: The distribution of covariates among the propensity-matched combined CEA/CABG and isolated CABG groups were similar. Among the propensity-matched pairs in the primary analysis, overall morbidity and mortality were higher in the combined CEA/CABG group compared with the CABG group alone (overall morbidity 15% v 8.8%, p = 0.025, and mortality 5.2% v 1.1%, p = 0.007, respectively). Median intensive care unit (ICU) length of stay was longer (47 v 31 hours, p = 0.004) and hospital length of stay was longer (12 v 9 days, p < 0.001) for the combined CEA/CABG surgery compared with isolated CABG surgery, respectively. Postoperative cardiac, neurologic, serious infection, and renal morbid events were similar between the 2 groups. In the secondary analysis, the rates of mortality, overall morbidity, and neurologic morbidity were similar between the groups, whereas the median ICU and hospital length of stay were significantly longer in the combined CEA/CABG group (47.6 v 39.8 hours, p = 0.025, and 12.0 v 9.0 days, p < 0.001, respectively). CONCLUSIONS: Increased mortality and overall morbidity outcomes were found in the combined CEA/CABG group when compared with well-matched isolated CABG patients, but similar when compared with well-matched isolated CABG patients with a history of previous CEA. Patients undergoing combined CEA/CABG procedures had significantly longer ICU and hospital lengths of stay compared with patients undergoing isolated CABG procedures. PMID- 17138084 TI - Staged anesthesia for combined carotid and coronary artery revascularization: a different approach. AB - OBJECTIVE: Combined coronary artery bypass graft (CABG) surgery and carotid endarterectomy (CEA) are performed in an attempt to reduce the risk of postoperative stroke after CABG surgery in patients with significant or symptomatic carotid artery stenosis. The choice between regional and general anesthesia for CEA is still under debate. Regional anesthesia offers an excellent monitoring technique of the neurologic status of the awake patient during carotid clamping. In an attempt to improve monitoring of the neurologic status and avoid the use of temporary shunting in patients undergoing the combined procedure, a different approach is described combining regional anesthesia for CEA followed immediately by general anesthesia for CABG surgery. DESIGN: Prospective nonrandomized case series. SETTING: University hospital. PARTICIPANTS: Twenty patients scheduled for combined CEA and CABG surgery underwent a "staged" anesthetic approach from January to December 2004. INTERVENTIONS: Pulmonary, femoral artery, and urinary catheters were inserted under local anesthesia. A deep cervical plexus block was then performed and supplemented by a superficial cervical plexus block. The patient was draped for standard combined CEA and CABG surgery. CEA was then performed using standard techniques. Without altering the surgical field, general anesthesia was given and endotracheal intubation performed following the successful CEA. Coronary revascularization was then completed. MEASUREMENTS AND MAIN RESULTS: CEA and CABG surgery were completed successfully in all patients. There was no need for conversion from local to general anesthesia. Endotracheal intubation was easily performed in all patients. There was no hospital mortality in this series. No neurologic events were observed during the CEA. A reversible ischemic stroke, ipsilateral to the CEA, occurred postoperatively on awakening from CABG surgery in 1 patient. CONCLUSIONS: This staged anesthetic approach for combined CABG and CEA surgery is an alternative in this complex subset of patients. PMID- 17138085 TI - Calcium supplementation of saline-based colloids does not produce equivalent coagulation profiles to similarly balanced salt preparations. AB - OBJECTIVES: The primary objective of this study was to test the hypothesis that calcium alone does not account for the observed coagulation differences between saline-based and balanced electrolyte IV fluid preparations. DESIGN: Controlled, nonblinded, in vitro observational study. SETTING: University-based anesthesia research laboratory. PARTICIPANTS: Ten healthy volunteers. INTERVENTIONS: The volunteers donated fresh blood for in vitro 40% and 60% dilution with 6 intravenous fluid preparations (lactated Ringer's solution, human albumin solution, and 4 hydroxyethyl starch preparations). All saline-based fluids were supplemented with calcium chloride to obtain ionized concentrations >or=1.0 mmol/L. MEASUREMENTS AND MAIN RESULTS: After dilution of the fresh blood with the study fluids, samples were analyzed by using the Thrombelastograph. Three colloid preparations produced minimal coagulation derangement, even at 60% dilution (human albumin solution, tetrastarch in saline, and pentastarch in balanced electrolyte solution), whereas pentastarch in saline and hetastarch in balanced electrolyte produced a mildly hypocoagulable state at 60% dilution. CONCLUSIONS: The different coagulation profiles between the 2 pentastarch preparations, as well as similar profiles of pentastarch in saline and hetastarch in balanced electrolyte solution, suggest that calcium is not solely responsible for previously observed effects. PMID- 17138086 TI - Comparison of hemodynamic changes after acute normovolemic hemodilution using Ringer's lactate versus 5% albumin in patients on beta-blockers undergoing coronary artery bypass surgery. AB - OBJECTIVE: Acute normovolemic hemodilution (ANH) is used cautiously in coronary artery disease (CAD) patients because of concerns of compromised coronary blood flow. This study aimed to compare hemodynamic changes by using either Ringer's lactate or albumin for ANH in CAD patients receiving beta-blockers. DESIGN: Prospective, randomized study. SETTING: Postgraduate teaching hospital. PARTICIPANTS: Thirty patients undergoing coronary artery bypass graft surgery (CABG) (hemoglobin >12 g/dL, on chronic beta-blocker therapy). INTERVENTIONS: Monitoring, induction, and anesthesia followed a routine protocol for CABG surgery including pulmonary artery catheter placement. Patients were randomly included in group 1 (ANH by Ringer's lactate) or in group 2 (ANH by 5% albumin). A hemodynamic calculation software program was used for parameters recorded before and after ANH. MEASUREMENTS AND MAIN RESULTS: ANH could not be completed in 5 patients (33%) in group 1 because of a fall in mean arterial pressure (MAP) of more than 25% from baseline. In both groups posthemodilution MAP, heart rate, systemic vascular resistance, and oxygen delivery index decreased, whereas stroke volume index, cardiac index, and tissue oxygen extraction increased significantly as compared to baseline values (p < 0.05). Hemodynamic parameters were better maintained during the study period in group 2 than group 1. CONCLUSIONS: Hemodynamic stability was better maintained by 5% albumin than Ringer's lactate for ANH in chronic beta-blocked CAD patients. Despite an increase in cardiac index, systemic oxygen delivery was decreased irrespective of the hemodiluting fluid used. ANH to a hemoglobin value of 10 g/dL in chronically beta-blocked CAD patients was well tolerated. PMID- 17138087 TI - Correlation between cerebral and mixed venous oxygen saturation during moderate versus tepid hypothermic hemodiluted cardiopulmonary bypass. AB - OBJECTIVE: This study was undertaken to compare cerebral oxygen saturation (RsO(2)) and mixed venous oxygen saturation (SvO(2)) in patients undergoing moderate and tepid hypothermic hemodiluted cardiopulmonary bypass (CPB). DESIGN: Prospective study. SETTINGS: University hospital operating room. PARTICIPANTS: Fourteen patients undergoing elective coronary artery bypass graft surgery using hypothermic hemodiluted CPB. INTERVENTIONS: During moderate (28 degrees -30 degrees C) and tepid hypothermic (33 degrees -34 degrees C) hemodiluted CPB, RsO(2) and SvO(2) were continuously monitored with a cerebral oximeter via a surface electrode placed on the patient's forehead and with the mixed venous oximeter integrated in the CPB machine, respectively. MEASUREMENTS AND MAIN RESULTS: Mean +/- standard deviation of RsO(2), SvO(2), PaCO(2), and hematocrit were determined prebypass and during moderate and tepid hypothermic phases of CPB while maintaining pump flow at 2.4 L/min/m(2) and mean arterial pressure in the 60- to 70-mmHg range. Compared with a prebypass value of 76.0% +/- 9.6%, RsO(2) was significantly decreased during moderate hypothermia to 58.9% +/- 6.4% and increased to 66.4% +/- 6.7% after slow rewarming to tepid hypothermia. In contrast, compared with a prebypass value of 78.6% +/- 3.3%, SvO(2) significantly increased to 84.9% +/- 3.6% during moderate hypothermia and decreased to 74.1% +/ 5.6% during tepid hypothermia. During moderate hypothermia, there was poor agreement between RsO(2) and SvO(2) with a gradient of 26%; however, during tepid hypothermia, there was a strong agreement between RsO(2) and SvO(2) with a gradient of 6%. The temperature-uncorrected PaCO(2) was maintained at the normocapnic level throughout the study, whereas the temperature-corrected PaCO(2) was significantly lower during the moderate hypothermic phase (26.8 +/- 3.1 mmHg) compared with the tepid hypothermic phase (38.9 +/- 3.7 mmHg) of CPB. There was a significant and positive correlation between RsO(2) and temperature-corrected PaCO(2) during hypothermia. CONCLUSIONS: During moderate hypothermic hemodiluted CPB, there was a significant increase of SvO(2) associated with a paradoxic decrease of RsO(2) that was attributed to the low temperature-corrected PaCO(2) values. During tepid CPB after slow rewarming, regional cerebral oxygen saturation was increased in association with an increase with the temperature corrected PaCO(2) values. The results show that during hypothermic hemodiluted CPB using the alpha-stat strategy for carbon dioxide homeostasis, cerebral oxygen saturation is significantly higher during tepid than moderate hypothermia. PMID- 17138088 TI - Assessment of cerebral oxygen balance during deep hypothermic circulatory arrest by continuous jugular bulb venous saturation and near-infrared spectroscopy. AB - OBJECTIVE: The purpose of this study was to compare jugular venous bulb saturation (SjvO(2)) and regional cerebral oximetry (rSO(2)) by near-infrared spectroscopy (NIRS) during procedures with deep hypothermic circulatory arrest (DHCA). DESIGN: Prospective observational study. SETTING: Academic hospital. PARTICIPANTS: Patients undergoing aortic reconstructive surgery with DHCA from July 2001 to January 2005. INTERVENTION: The authors examined cerebral oxygenation by continuous NIRS monitoring and by blood gas analysis of intermittently sampled jugular bulb blood (SjvO(2)). Data were obtained during various stages of the procedure in 29 patients. NIRS measurements were compared with SjvO(2). MEASUREMENTS AND MAIN RESULTS: NIRS and SjvO(2) trends were similar. Overall, cerebral venous oxygen saturation obtained from NIRS was lower compared with SjvO(2) (p < 0.05), especially during periods of low temperature. The mean correlation between NIRS and SjvO(2) was 0.363, and the individual correlations varied from -0.11 to 0.91. The low mean correlation was because of a high degree of variability in the NIRS data between patients. CONCLUSION: It was concluded that NIRS does not closely correlate with SjvO(2) in this patient population. Cerebral oximetry measured by NIRS could not replace jugular bulb saturation as an intraoperative marker of adequate metabolic suppression. PMID- 17138089 TI - Left main coronary artery spasm: medical versus surgical management. PMID- 17138090 TI - Acute spontaneous coronary artery dissection in the peripartum period. PMID- 17138091 TI - Mitral valve repair: to slide or not to slide-precardiopulmonary bypass echocardiogram examination. PMID- 17138092 TI - Implications of intraoperative transesophageal echocardiography detection of ruptured sinus of valsalva in a patient with severe aortic regurgitation undergoing aortic valve replacement. PMID- 17138093 TI - Combined carotid endarterectomy and off-pump coronary artery bypass surgery under thoracic epidural anesthesia without endotracheal general anesthesia. PMID- 17138094 TI - Pseudoaneurysm of the innominate artery: a delayed iatrogenic complication after internal jugular vein catheterization. PMID- 17138095 TI - Accessory tricuspid valve tissue in tetralogy of fallot causes hemodynamic changes during intermittent positive-pressure ventilation. PMID- 17138096 TI - Retroperitoneal hematoma: an unusual complication after femoral vein cannulation. PMID- 17138097 TI - Intraoperative dobutamine stress echocardiography to assess aortic valve stenosis. PMID- 17138098 TI - Dobutamine stress echocardiography and intraoperative assessment of mitral valve. PMID- 17138099 TI - Anesthesia implications of blast injury. PMID- 17138100 TI - Case 6-2006. Percutaneous superior vena cava cannulation for repeat sternotomy in cardiac operations. PMID- 17138101 TI - Pro: Antifibrinolytics are safe and effective in patients undergoing liver transplantation. PMID- 17138102 TI - Con: Antifibrinolytics are not safe and effective in patients undergoing liver transplantation. PMID- 17138103 TI - Transesophageal echocardiographic bull's eye. PMID- 17138104 TI - Unusual images of perioperative transesophageal echocardiography. PMID- 17138105 TI - Intraoperative transesophageal ultrasonography can measure left renal blood flow. PMID- 17138107 TI - Severe acute tricuspid regurgitation after the withdrawal of an entrapped pulmonary artery catheter. PMID- 17138108 TI - Echocardiography in suspected acute type a aortic dissection: detection and definitive management of a false-positive presentation. PMID- 17138109 TI - Evaluation of the supraglottic retractor for guided intraopertive transesophageal echocardiography probe insertion. PMID- 17138110 TI - Fluid management in cardiac surgery: is one hydroxyethyl starch solution safer than another? PMID- 17138111 TI - What can we learn from success? PMID- 17138112 TI - Breast specimen radiography: can it predict margin status of excised breast carcinoma? AB - RATIONALE AND OBJECTIVES: This study examined the value of intraoperative specimen radiography (SR) in determining margin status of excised breast lesions. Of interest was the concordance between the radiologic and histopathologic interpretation of margins. We investigated the influence of in situ disease and of one versus two radiologic views on this concordance. MATERIALS AND METHODS: Our study consisted of 112 women who underwent breast conservation therapy (BCT) during 2002. Margins were examined with one- or two-view SR. Margins were histologically positive if malignant cells resided < or = 1 mm from the specimen edge. The McNemar's test was used to determine concordance between SR and histopathology (HP). Because surgeons excised extra tissue in cases of positive radiologic margins, we believe that a change in margin status occurred in which true positives became false positives. Accordingly, we analyzed our data with multiple iterations in which, one by one, false positives were considered true positives. RESULTS: Concordance between SR and HP reached statistical significance after 5/17 false positives were considered true positives. Data excluding DCIS reached significance after 6 of 6 false positives were considered true positives. One- and two-view SR reached significance when 2 of 8 and 7 of 9 false positives, respectively, were considered true positives. CONCLUSION: In conclusion, our study suggests that SR can aid in margin assessment for patients undergoing BCT. We did not find that concordance between SR and HP is higher in cases of purely invasive disease. Concordance was higher in one-view SR in comparison to two-view. A larger sample size should be analyzed before recommending against using two views. PMID- 17138113 TI - Mapping of subcortical white matter abnormality in Alzheimer's disease using diffusion-weighted magnetic resonance imaging. AB - RATIONALE AND OBJECTIVES: White matter (WM) abnormality in Alzheimer's disease (AD) has been less well characterized than cortical damage. We studied the spatial distribution of the subcortical WM abnormality using diffusion-weighted magnetic resonance imaging (DWI). MATERIALS AND METHODS: Twenty-one AD patients and seven healthy, elderly subjects were included. DWIs were obtained using a cerebrospinal fluid (CSF)-nulled pulse sequence to reduce the partial volume contamination of CSF signal. Diffusivity in the subcortical WM voxels was mapped onto the cortical surface using original software so that the spatial distribution of subcortical WM damage, which was visualized as an area of increased diffusivity, could be viewed in a three-dimensional map. The damages in the lateral surface of the bilateral cerebral hemispheres were visually evaluated, and severities of the damages in five brain regions were compared with each other. In addition, the severity of the damage in each region was correlated with patient's mini-mental state examination (MMSE) score. RESULTS: In both hemispheres, clear sparing of the pericentral regions and predominant involvement of the parietal and temporal regions were revealed with statistical significance (P < .05, respectively). Marginal correlation (P < .05 uncorrected for multiple comparisons) was observed between the damage severity in the bilateral frontal and right temporal regions and patient's MMSE score. CONCLUSION: We demonstrated a subcortical WM abnormality over the parietal and temporal regions with clear sparing of the pericentral region using our mapping method, which supported the hypothesis that the subcortical WM abnormality in AD originates in Wallerian degeneration. PMID- 17138114 TI - Assessment of coronary artery stent patency and restenosis using 64-slice computed tomography. AB - RATIONALE AND OBJECTIVES: Restenosis remains a major limitation of coronary catheter-based stent placement. Therefore, a reliable noninvasive diagnostic method for the evaluation of stented coronary arteries would be highly desirable. Our aim was to evaluate the diagnostic accuracy of high-resolution 64-slice computed tomography (64SCT) in a pilot study for the assessment of the lumen of coronary artery stents. MATERIALS AND METHODS: Twenty-five patients underwent 64SCT of the coronary arteries and quantitative x-ray coronary angiography (QCA) after coronary artery stent placement. 64SCT coronary angiography was performed with the following parameters: spatial resolution = 0.4 x 0.4 x 0.4 mm; temporal resolution = 83-165 milliseconds; contrast agent = 80 mL at a flow rate of 5 mL/second; retrospective electrocardiogram gating. The 64SCT scans were evaluated for image quality and for the presence of significant in-stent and peri-stent (proximal and distal) stenoses. Determinations were made of the sensitivity, specificity, diagnostic accuracy, and positive and negative predictive values (PPV and NPV) of 64SCT for the detection or exclusion of stenoses. RESULTS: A total of 46 stents were evaluated, of which 45 (98%) were of diagnostic image quality. Significant in-stent restenosis or occlusion was detected on QCA in 8/45 cases (>/=50% stenosis = 6; occlusion = 2). The sensitivity, specificity, accuracy, PPV, and NPV of 64SCT for the detection of significant in-stent disease was 75%, 92%, 89%, 67%, and 94%, respectively. Both occluded coronary artery stents were correctly identified. The sensitivity, specificity, and accuracy values of 64SCT for the detection of significant proximal peri-stent stenoses were 75%, 95%, and 93%, respectively, whereas the values for detection of significant distal peri-stent stenoses were 67%, 85%, and 84%, respectively. CONCLUSION: The high spatial and temporal resolution of 64SCT may permit improved assessment of stent occlusion and peri-stent disease, although detection of in stent stenosis remains difficult. PMID- 17138115 TI - Identification, segmentation, and image property study of acute infarcts in diffusion-weighted images by using a probabilistic neural network and adaptive Gaussian mixture model. AB - RATIONALE AND OBJECTIVES: Accurate identification of infarcted regions of the brain is critical in management of stroke patients. An efficient and fast method for identification and segmentation of infarcts in the diffusion-weighted images (DWI) is proposed. MATERIALS AND METHODS: Thirteen stroke patients were studied. DWI scans were acquired with a slice thickness of 5 mm. We have used a probabilistic neural network for selecting infarct slices and an adaptive (two level) Gaussian mixture model for segmentation of the infarcts. Statistical analysis, such as identification of distribution, first-order statistics calculation, and receiver operating characteristic curve analysis, was performed. RESULTS: The average dice index is about 0.6, and average sensitivity and specificity are about 81% and 99%, respectively. The value of sensitivity and dice index are influenced by the number of false positives and false negatives. Because artifacts and infarcts have similar imaging characteristics, it is difficult to completely eliminate the artifacts. The accuracy of localization is nearly 100% as there were only two false-positive and three false-negative slices of all 381 slices. The algorithm takes about 1 minute in the Matlab computing environment to process a volume. CONCLUSION: A method to localize and segment the acute brain infarcts is proposed. The method aids the clinician in reducing the time needed to localize and segment the infarcts. The speed of localization and segmentation can be enhanced further by implementing the algorithm in VC++ and using fast algorithms for selection of Gaussian mixture model parameters. PMID- 17138116 TI - Clinical manifestations and MRI findings of patients with hydrated and dehydrated lumbar disc herniation. AB - RATIONALE AND OBJECTIVES: In addition to the expected appearance of degenerated discs that become dehydrated, hydrated intervertebral disc herniations are sometimes encountered in radiologic practice. This study was undertaken to evaluate the clinical manifestations and MRI findings of hydrated and dehydrated herniated intervertebral discs. MATERIALS AND METHODS: This cross-sectional single-institution study included 73 patients with dehydrated (group I) and hydrated (group II) lumbar disk herniation. The criteria for hydrated and dehydrated disc herniation were subjective criteria compared to the normal signal of intervertebral discs. A herniated disc has been regarded to be hydrated if more than two thirds of it was hypersignal in T2-weighted images, while more than two thirds of a dehydrated disc was hyposignal on T2-weighted images. RESULTS: The mean weight of patients in group I was greater than that of patients in group II (69.3 versus 64.2 kg, P < 0.05). Also, patients in group I tended to be older than those in group II (35.2 versus 28.9 years). Regarding physical activity, a greater number of patients in group II had intense physical activity compared to group I patients (25% versus 13.2%, respectively; P < 0.05). The duration of radicular pain and back pain was significantly greater in group I than in group II (485 versus 202 and 1346 versus 242 days, respectively; P < 0.05). CONCLUSION: Hydrated intervertebral disc herniation tends to be associated with younger age, lighter body weight, shorter duration of radicular pain, and more intense physical activity compared to dehydrated intervertebral disc herniation. These findings may suggest other mechanisms rather than degenerative changes for hydrated disc herniation. PMID- 17138117 TI - Assessment of polyp and mass histopathology by intravenous contrast-enhanced CT colonography. AB - RATIONALE AND OBJECTIVES: We sought to demonstrate that intravenous contrast enhanced CT colonography (CTC) can distinguish colonic adenomas from carcinomas. METHODS: Supine intravenous contrast-enhanced CTC with colonoscopic and/or surgical correlation was performed on 25 patients with colonic adenomas or carcinomas. Standard deviation of mean polyp CT attenuation was computed and assessed using ANOVA and receiver-operating characteristic analyses. RESULTS: Colonoscopy confirmed 32 polyps or masses 1 to 8 cm in size. The standard deviations of CT attenuation were carcinomas (n = 13; 36 +/- 6 HU; range 28-48 HU) and adenomas (n = 19; 49 +/- 14 HU; range 31-100 HU) (P = 0.005). At a standard deviation threshold of 42 HU, the sensitivity and specificity for classifying a polyp or mass as a carcinoma were 92% and 79%, respectively. The area under the receiver-operating characteristic curve was 0.89 +/- 0.06 (95% confidence interval 0.73-0.96). CONCLUSIONS: Measurement of the standard deviation of CT attenuation on intravenous contrast-enhanced CTC permits histopathologic classification of polyps 1 cm or larger as carcinomas versus adenomas. The presence of ulceration or absence of muscular invasion in carcinomas creates overlap with adenomas, reducing the specificity of carcinoma classification. PMID- 17138118 TI - Real-time sonoelastography performed in addition to B-mode ultrasound and mammography: improved differentiation of breast lesions? AB - RATIONALE AND OBJECTIVES: The goal of the present study was to compare the sensitivity and specificity of elastography with that of B-mode ultrasound (US) and mammography. MATERIALS AND METHODS: A total of 300 patients with histologically confirmed breast lesions (168 benign, 132 malignant) were included. Evaluation was by means of the three-dimensional finite-element method. The data are color-coded and superimposed on the B-mode US scan. The images were evaluated by two independent readers. The results were compared with mammography, histology, and the data obtained by previous US investigations. Sensitivities and specificities were calculated. RESULTS: Sensitivity and specificity in the differentiation of benign and malignant lesions were 87% and 85%, respectively, for mammography and 94% and 83% for B-mode US. The two examiners were in very good agreement in their evaluation of the elastograms (kappa: 0.86). Elastography had a sensitivity of 82% and a specificity of 87%. Elastography was superior to B mode US in diagnosing Breast Imaging Reporting and Data System (BI-RADS) 3 lesions (92% vs. 82% specificity) and in lipomatous involution (80% vs. 69% specificity). CONCLUSION: Elastography in breast lesions showed a higher specificity and a lower sensitivity in comparison with B-mode sonography. Elastography may be beneficial in BI-RADS 3 lesions and in lipomatous involution. PMID- 17138119 TI - Quantification of bleomycin-induced murine lung damage in vivo with micro computed tomography. AB - RATIONALE AND OBJECTIVES: We explored noninvasive, in vivo cone-beam microcomputed tomography (micro-CT) to visualize and quantify fibrotic and inflammatory damage over the entire lung volume of mice. MATERIALS AND METHODS: We used bleomycin to induce pulmonary damage in vivo and compared the results from micro-CT with histologic measurements. Ten C57BL/6 mice were given 5 U/kg bleomycin intratracheally. Seven surviving mice were scanned with micro-CT before administration of bleomycin, and again before sacrifice. The resulting images were analyzed for lung volume measurements. After the final scan, all lungs were examined histologically and pulmonary damage was quantified. Damaged lung tissue regions were matched between micro-CT images and histologic sections for each mouse. RESULTS: The percent lung damage calculated from micro-CT and histology were correlated (r(2) = 0.49, r = 0.64 with P = 0.12), and the means of their respective distributions were not different (P > 0.05). CONCLUSION: This study shows that micro-CT is a promising alternative to predicting lung damage caused by bleomycin. CT image volumes of the thorax allow for global tissue sampling, which may be useful when following nonuniform lung damage that can occur from intratracheal administration of bleomycin. PMID- 17138121 TI - In vivo 1H MR spectroscopy in the evaluation of the serial development of hepatocarcinogenesis in an experimental rat model. AB - RATIONALE AND OBJECTIVES: We used a 1.5-T MR scanner to investigate in vivo hydrogen 1 ((1)H) MRS to evaluate metabolic changes in the hepatocarcinogenesis experimental rat model. MATERIALS AND METHODS: Hepatocellular carcinoma (HCC) was induced by diethylnitrosamine in 70 treated rats with 20 normal rats used as controls. Single-voxel (1)H MRS is performed to obtained the relative choline-to lipid (Cho/lipid) ratio. The liver and tumor tissues are incised for the histologic examination. Based on the histologic result, the progression of hepatocarcinogenesis of the animal model was divided into three stages: fibrosis stage, cirrhosis stage, and HCC stage. The mean (+/-SD) ratio values are calculated and compared at various stages between the treated group and the control group. RESULTS: In control group, the calculated mean (+/-SD) Cho/lipid ratio was 0.15 +/- 0.05. With the progression of hepatocarcinogenesis, the Cho/lipid ratio increased significantly, to 0.18 +/- 0.05, 0.24 +/- 0.07, and 0.38 +/- 0.19, respectively. CONCLUSION: The (1)H MRS is technically feasible for evaluation of the metabolic changes in the animal model. A significant increase in choline-containing compounds level was observed in the HCC stage in the treated group. PMID- 17138120 TI - Current status of CT colonography. PMID- 17138122 TI - Self-test software for PowerPoint: a tool for self-learning. AB - RATIONALE AND OBJECTIVES: We developed self-test software to improve self learning efficiency using Microsoft PowerPoint data files. CONCLUSION: This tool can be run on IBM-compatible computer under Microsoft Windows. It is a new useful and interactive tool for self-learning. This tool allows users to do view the cases in the PowerPoint data files by random or sequentially. Goal-oriented effective self-learning is possible from methods that conjecture the possible differential diagnosis without promptly seeing correct diagnosis. Thus effective and interactive self-learning is possible. PMID- 17138123 TI - Analysis of a mammography teaching program based on an affordance design model. AB - RATIONALE AND OBJECTIVES: The wide use of computer technology in education, particularly in mammogram reading, asks for e-learning evaluation. The existing media comparative studies, learner attitude evaluations, and performance tests are problematic. Based on an affordance design model, this study examined an existing e-learning program on mammogram reading. MATERIALS AND METHODS: The selection criteria include content relatedness, representativeness, e-learning orientation, image quality, program completeness, and accessibility. A case study was conducted to examine the affordance features, functions, and presentations of the selected software. Data collection and analysis methods include interviews, protocol-based document analysis, and usability tests and inspection. Also some statistics were calculated. RESULTS: The examination of PBE identified that this educational software designed and programmed some tools. The learner can use these tools in the process of optimizing displays, scanning images, comparing different projections, marking the region of interests, constructing a descriptive report, assessing one's learning outcomes, and comparing one's decisions with the experts' decisions. Further, PBE provides some resources for the learner to construct one's knowledge and skills, including a categorized image library, a term-searching function, and some teaching links. Besides, users found it easy to navigate and carry out tasks. The users also reacted positively toward PBE's navigation system, instructional aids, layout, pace and flow of information, graphics, and other presentation design. CONCLUSION: The software provides learners with some cognitive tools, supporting their perceptual problem solving processes and extending their capabilities. Learners can internalize the mental models in mammogram reading through multiple perceptual triangulations, sensitization of related features, semantic description of mammogram findings, and expert-guided semantic report construction. The design of these cognitive tools and the software interface matches the findings and principles in human learning and instructional design. Working with PBE's case-based simulations and categorized gallery, learners can enrich and transfer their experience to their jobs. PMID- 17138124 TI - The (not so) good old days. PMID- 17138125 TI - Why are meetings. PMID- 17138126 TI - Survival results in patients with screen-detected prostate cancer versus physician-referred patients treated with radical prostatectomy: early results. AB - OBJECTIVE: Screening using a standardized protocol may improve outcomes of patients undergoing treatment for prostate cancer. We compared the 7- year progression-free survival rates after radical retropubic prostatectomy in patients whose prostate cancer was detected through a formal screening program with those of patients referred for treatment by other physicians who did not use a standardized screening/referral protocol. METHODS: A single surgeon (W.J.C.) performed radical retropubic prostatectomy in 3,177 consecutive patients between 1989 and 2003. Of these patients, 464 had cancer detected in a screening study, and 2,713 were referred from outside institutions. We compared the screened and referred cohorts for age at surgery, clinical stage, pathologic stage, Gleason sum, preoperative prostate-specific antigen (PSA) levels, and adjuvant radiation therapy. Kaplan-Meier product limit estimates were used to calculate 7-year progression-free probabilities, and Cox proportional hazards models were used to determine the clinical and pathologic parameters associated with cancer progression in each group. RESULTS: The overall 7-year progression-free survival rates were 83% for the screened patients compared with 77% for the referred patients (P = 0.002). Preoperative PSA, Gleason sum, clinical stage, pathologic stage, and adjuvant radiotherapy were all significantly associated with cancer progression. There was a significantly higher proportion of referred patients with a preoperative PSA > or =10, Gleason sum > or =7, and nonorgan-confined disease. CONCLUSIONS: Patients with screened-detected prostate cancer have more favorable clinical and pathologic features, and 7-year progression-free survival rates than referred patients. On multivariate analysis, including other clinical variables, screening status was a significant independent predictor of biochemical outcome. PMID- 17138127 TI - Quality of life 2 years after salvage cryosurgery for the treatment of local recurrence of prostate cancer after radiotherapy. AB - PURPOSE: Previous research has raised concerns that although salvage cryosurgery may be an effective treatment to prevent the progression of prostate cancer after radiotherapy failure, the quality of life cost many be so severe as to prevent its acceptance as a viable treatment. The present study's purpose was to further the understanding of the quality of life outcomes of salvage cryosurgery. MATERIALS AND METHODS: A total of 46 men with locally recurrent prostate cancer after radiotherapy were recruited to participate in a prospective Phase II clinical trial using salvage cryosurgery. There were 2 questionnaires (i.e., the European Organization of Research and Treatment of Cancer QLQ C30 and the Prostate Cancer Index) administered before cryosurgery, and at 1.5, 3, 6, 12, 18, and 24 months after treatment. RESULTS: Quality of life returned to preoperative levels by 24 months after cryosurgery in all domains, with the exception of urinary and sexual functioning. At 24 months, 29% of men reported urinary bother as a moderate-to-big problem, and 56% reported sexual bother as a moderate-to-big problem. CONCLUSIONS: To our knowledge, this is the first study to evaluate prospectively men's quality of life for 2 years after salvage cryosurgery for locally recurrent prostate cancer after radiotherapy. Long-term impairments in quality of life appear to be limited to the sexual and urinary function domains. Overall quality of life appears to be high. These results support salvage cryosurgery as a viable treatment option. PMID- 17138128 TI - Phase II study of capecitabine single-agent therapy in patients with metastatic renal cell carcinoma. AB - Fluoropyrimidines are known to have modest activity in the treatment of metastatic renal cell carcinoma (RCC). Capecitabine is an orally administered prodrug that is converted to fluorouracil and is of potential use in the treatment of this disease. We conducted a Phase II clinical trial of capecitabine administered as a single agent to patients with metastatic RCC. The treatment consisted of 1250 mg/m(2) capecitabine orally, twice daily (2500 mg/m(2) per day) days 1-14, repeated every 21 days. There were 15 patients, including 13 men and 2 women, who underwent a total of 67 cycles (median 3.5; range 1-15). Nine patients had undergone prior systemic therapy consisting of interferon-alpha in 3, interleukin-2 in 1, interferon-alpha plus interleukin-2 in 4, and investigational therapy with bryostatin-1 in 1. There were 14 patients assessable for response (one withdrew), and no responses were seen. Median time to progression was 9 weeks (range 1-45). There were 3 patients (21%) who had stable disease for 18, 39, and 45 weeks. Hematologic toxicity was mild. Three patients had grade 3 or 4 gastrointestinal toxicity, and 3 required dose reductions. There were 2 early deaths, including 1 patient with pulmonary edema and 1 with hypotension. The study was terminated because there were no responses in the first 14 assessable patients, indicating that the response rate was likely to be less than 20%. We conclude that single-agent capecitabine has minimal activity for the treatment of metastatic RCC. PMID- 17138129 TI - Improved prostate cancer detection with anterior apical prostate biopsies. AB - PURPOSE: Research to improve prostate cancer detection with transrectal ultrasound-guided prostate biopsies has focused on increasing the number of cores and the directing of biopsies laterally. In this study, we describe our experience with the addition of anterior apical biopsies. MATERIALS AND METHODS: A total of 164 consecutive patients with an increased or increasing prostate specific antigen and/or abnormal digital rectal examination underwent transrectal ultrasound and systematic biopsy. We performed our standard laterally directed sextant biopsies plus additional mid parasagittal plane biopsies at the base and mid-gland, and an anteriorly directed biopsy at the apex. Site-specific detection and tumor characteristics are reported. RESULTS: Prostate cancer was detected in 71 patients (43.3%). The most commonly unique site was the anterior apex. Excluding these biopsies would have missed 17% of the cancers detected. The cancers limited to the anterior apex had tumor characteristics similar to all other cancers detected. CONCLUSION: In our experience, the anterior apical biopsies increase the detection of prostate cancer on transrectal ultrasound guided biopsies. Further study on incorporating this site into the biopsy scheme is indicated. PMID- 17138130 TI - E-cadherin promoter polymorphisms are not associated with the aggressiveness of prostate cancer in Caucasian patients. AB - BACKGROUND: -160C-->A and -347G-->GA polymorphisms in the promoter region decrease E-cadherin gene transcription. Decreased E-cadherin expression predicts poor outcome among patients with cancer. We sought to investigate whether -160C- >A and/or -347G-->GA polymorphisms were associated with the aggressiveness of prostate cancer. METHODS: TaqMan single nucleotide polymorphism genotyping assay (Applied Biosystems, Foster City, CA) was used to detect -160C-->A and -347G-->GA polymorphisms in deoxyribonucleic acid from the paraffin-embedded prostate tissues of 98 Caucasian patients. RESULTS: The genotype frequencies were -160C/C: 48% (47 of 98); -160C/A: 44% (43 of 98); -160A/A: 8% (8 of 98); -347G/G: 68% (67 of 98); -347G/GA: 28% (27 of 98); and -347GA/GA: 4% (4 of 98). Using the chi square test, we found that the polymorphisms -160C-->A and -347G-->GA were not related to other clinical and pathologic parameters (i.e., age, prostate-specific antigen level, Gleason grade, and clinical stage) (P > 0.05). In combination analysis, there was no significant relationship between patients with both 160C/C and -347G/G, and these same parameters (P > 0.05). Using the log-rank test, we found no significant difference in relapse-free survival and overall survival between patients with -160C/C and those with -160A/C or -160A/A (P = 0.0764 and 0.2746, respectively), and also no significant difference between patients with -347G/G and those with -347GA/G or -347GA/GA (P = 0.9416 and 0.7367, respectively). There was also no significant difference in relapse-free survival and overall survival between patients with homozygosities of -160C/-347G and patients with other genotypes (P = 0.1418 and 0.2434, respectively). CONCLUSION: We conclude that E-cadherin -160C-->A and/or -347G-->GA polymorphisms are not associated with the aggressiveness of prostate cancer in Caucasian patients. PMID- 17138131 TI - Treatment of a radiation-induced rectal ulcer with hyperbaric oxygen therapy in a man with prostate cancer. AB - Late radiation proctopathy is a painful and vexing complication of prostate radiation. We report a case of a 55-year-old man with prostate cancer, and complaints of tenesmus and severe rectal pain after radiation therapy. The patient was diagnosed with a locally advanced Gleason score 8 prostate cancer and an increased prostate-specific antigen of 42.3 ng/ml. His past medical history was notable for a history of bilateral lymph node dissection complicated by Clostridium difficile colitis. He subsequently received 3-dimensional conformal radiation therapy. Seven months after completing therapy, minor rectal bleeding and significant pain developed, requiring increasing doses of opioid analgesics. Fourteen months after 3-dimensional conformal radiation therapy, sigmoidoscopy revealed a single chronic deep ulcer at the anorectal junction. As an alternative to diverting colostomy, the patient underwent a course of hyperbaric oxygen. Within 1 month of completing hyperbaric oxygen treatment, his symptoms completely resolved. Nine months from completion of hyperbaric oxygen therapy, he has had no recurrence of symptoms. Hyperbaric oxygen therapy can be considered a treatment option after failure of standard treatments in patients with severe radiation proctopathy. PMID- 17138133 TI - Nucleic acid-based marker approaches to urologic cancers. AB - There are numerous molecular modifications known to occur in cancer. New nucleic acid-based biomarkers provide a unique approach to patient management in urologic oncology. Malignant transformation of a normal cell requires a series of epigenetic and genetic changes or "hits." Epigenetics produced by deoxyribonucleic acid methylation, adding a methyl group to the fifth position of cytosine within CpG dinucleotides, are important players in deoxyribonucleic acid repair, genome instability, and regulation of chromatin structure. Genetic alterations in cancer can include mutations, chromosome deletions, insertions, amplifications, and translocations. In addition, the modifications of telomeres are critical to the maintenance of chromatin structure, transcription, and cell function in cancer. We review only nucleic acid-based molecular biomarkers in urologic oncology that can assist the clinician in establishing the diagnosis of disease, or that can predict the behavior of the disease or the patient's survival. PMID- 17138134 TI - Urinary bladder tumor markers. AB - Bladder cancer is amenable to biomarker development because many tumor-associated molecules are secreted in urine. Tumor cells are shed in urine, and, therefore, tests that detect tumor cell-surface markers have also been developed to diagnose bladder cancer and monitor its recurrence. Several bladder tumor markers show higher sensitivity than cytology, but most have lower specificity. In addition to markers that use conventional technologies such as enzyme-linked immunosorbent assay, point-of-care devices, reverse transcriptase polymerase chain reaction, fluorescent in situ hybridization, and immunocytochemistry, proteomic and gene profiling approaches are being used to find new biomarkers to assist in the molecular profiling of bladder cancer. This review describes both new and well studied bladder tumor markers. PMID- 17138135 TI - Molecular markers of prostate cancer. AB - Although prostate-specific antigen (PSA) has evolved as a very useful tool for detection of prostate cancer, there remains an urgent need for more accurate biomarkers to diagnose prostate cancer and predict cancer-related outcomes. Recent advances in the study of proteomics and high throughput techniques have led to the discovery of many potential biomarkers for prostate cancer. This article briefly reviews the current status of PSA testing and discusses several candidate protein biomarkers for prostate cancer, as well as highlighting some recent proteomic discoveries with the potential to supplement or even replace PSA for the diagnosis and prognosis of prostate cancer. PMID- 17138156 TI - Cardiovascular effects of epinephrine under sedation with nitrous oxide, propofol, or midazolam. AB - OBJECTIVE: During implant surgery, a large amount of local anesthetics containing epinephrine are often required, and the resulting cardiovascular effects of administered epinephrine are not negligible. On the other hand, sedation has wide applications in implant surgery. Nitrous oxide, propofol, or midazolam are commonly used as sedative drugs, and each has also its own cardiovascular effects. The objective of this study was to investigate the cardiovascular effects of epinephrine on patients under sedation with nitrous oxide, propofol, or midazolam. STUDY DESIGN: We studied 9 healthy volunteers. They received epinephrine infusion at a nominal rate of 10, 25, or 50 ng/kg per minute under sedation with 30% nitrous oxide inhalation, 4 mg/kg per hour intravenous propofol or 0.2 mg/kg per hour intravenous midazolam. For each, hemodynamic response and blood pressure and heart rate variability were measured. RESULTS: When epinephrine was infused alone at 50 ng/kg per minute, heart rate (HR) and cardiac index (CI) increased by 19.5% and 40.7%, respectively. Propofol suppressed the epinephrine-induced increase in CI. During midazolam infusion, the highest dose of epinephrine caused a 37.5% increase in HR, which was significantly higher than for epinephrine infusion alone. This response was accompanied by the reduction in high-frequency power of heart rate variability, suggesting decreased parasympathetic activity. Nitrous oxide had no influence on the cardiovascular response to epinephrine. CONCLUSION: Increased cardiovascular activity due to epinephrine can be alleviated by propofol. However, midazolam and nitrous oxide are of no advantage for stabilizing the hemodynamic status of the patient. Intravenous sedation with propofol is useful during oral surgical procedures in which a large amount of epinephrine is required. PMID- 17138157 TI - In vivo tetracycline labeling of bone: an intraoperative aid in the surgical therapy of osteoradionecrosis of the mandible. AB - In this technical approach, we demonstrate that preoperative tetracycline administration helps distinguish between viable and necrotic bone in osteoradionecrosis (ORN) during surgery by exciting tetracycline fluorescence by black light. The difficult aspect of the operative approach lies in the decision of the extent of the debridement in order to prevent an iatrogenic fracture or progression of ORN. With this technique, vital bone, in contrast to necrotic bone, showed detectable fluorescence. Therefore, tetracycline fluorescence labeling may be used in attempts to facilitate the intraoperative decision making in the surgical therapy of osteoradionecrosis. PMID- 17138158 TI - Biochemical and histopathological findings of N-butyl-2-cyanoacrylate in oral surgery: an experimental study. AB - OBJECTIVES: The increasing use of cyanoacrylates in dentistry, particularly as an adhesive and sealing glue, has raised concerns regarding its potential toxicity in humans. Several different forms of these compounds including methyl- (MCA), ethyl- (ECA), isobutyl-, isohexyl-, and octyl CA have been developed to eliminate tissue toxicity. N-butyl-2-cyanoacrylate is becoming an increasingly popular method for wound closure under low tension. Despite their increasing use, pharmacologic effects of these substances on liver and kidney functions are not widely known. The objective of the present study was to investigate possible immediate and long-term systemic effects of N-butyl-2-cyanoacrylate in oral surgery. STUDY DESIGN: Ten male Wistar rats weighing 220 to 270 g were used in the study. Straight incisions were made to the buccal mucosa of the animals. N butyl-2-cyanoacrylate adhesive (Indermil) was applied and wounds were closed primarily. Blood specimens were taken periodically from the vena cava of the animals before the surgical procedure and 2, 14, 21, and 65 days after the surgical procedure. The blood specimens of those taken before the application of the adhesive were defined as the control group; blood specimens that were taken 2, 14, 21, and 65 days from the application were defined as study group. The stored plasma samples were analyzed for blood urea nitrogen (BUN), creatinine (CRE), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBI), total protein (TP), albumin (ALB), and amylase (AML). In addition to biochemical parameters, histopathological examination was performed. Blood parameter values of the control and study groups were statistically compared with the Duncan test (P < .05). RESULTS: There were no significant differences in the values of BUN, CRE, ALT, AST, TBI, TP, ALB, and AML between the control and at 2, 14, 21, and 65 days. CONCLUSION: The present study shows that N-butyl-2-cyanoacrylate is a suitable adhesive applicable in oral surgery. PMID- 17138159 TI - Delayed traumatic ocular emphysema: a case report. AB - A 47-year-old man sustained fractures of the left orbita and zygomatic bone after a fall. He was advised to visit an oral and maxillofacial department for more detailed evaluation and treatment, but ignored the advice. Four days later, after sneezing and blowing his nose, he presented severe exophthalmos, diplopia, and reduction of vision. Two days after the symptoms presented he asked for oral and maxillofacial surgery consultation. He refused any surgical intervention and was treated conservatively. The recovery was slow and 1 week later he was discharged with slight esthetic problems. PMID- 17138160 TI - Unexpected breakage of mandibular midline distraction device: case report. AB - Management of the transverse mandibular deficiency and anterior crowding by mandibular midline distraction osteogenesis (MMDO) is an efficient and stable alternative to orthodontic mechanics with minor complications. Although the only major complication reported previously during MMDO is the nonunion of the segments, in the present case report an unexpected breakage of the distractor in MMDO during the consolidation period as a new major complication was presented. PMID- 17138161 TI - Osteoma of maxillary sinus: a case report. AB - Craniofacial osteomas may be located either superficially or intraosseously on any bone of the cranium or face or within a paranasal sinus. Osteomas within the paranasal sinus account for 0.01% to 0.43% of patients; of these, up to 80% occur in the frontal sinus. An osteoma of maxillary sinus is extremely rare. We describe a rare case of osteoma of a 56-year-old Asian woman in the maxillary sinus. PMID- 17138162 TI - Vasovagal syncope with asystole associated with intravenous access. AB - Two cases of vasovagal syncope (VVS) during venous access are reported. Both patients had a history of fainting episodes and experienced bradycardia with asystole, hypotension, and fainting. Pain and phobic stress during venous access triggered an increase in parasympathetic tone, resulting in bradycardia with asystole and hypotension in both cases. Hypotension and bradycardia likely caused cerebral hypoperfusion, leading to fainting. The intense parasympathetic tone triggered by somatic or emotional stress was likely responsible for directly depressing the sinus node, leading to asystole and bradycardia. Bradycardia with asystole progressing to syncope is a potentially fatal dysrhythmia in patients with cardiovascular disease or older patients with decreased cardiac function. Appropriate treatment for VVS includes the administration of intravenous fluids, vagolytics, ephedrine, and the rapid use of the Trendelenburg position. Intravenous fluids and atropine were used to treat the present patients. PMID- 17138163 TI - Difficulties in anesthetic management of patients with micrognathia: report of a patient with Stickler syndrome. AB - Stickler syndrome is an autosomal dominant multisystem disorder with characteristic midface hypoplasia, retromicrognathia, cleft palate and a "moon shaped" appearance. Progressive myopia and retinal degeneration are frequent. It is estimated that one third of all Pierre Robin patients have Stickler syndrome. Patients with a mandibular hypoplasia like Stickler syndrome present the anesthesiologist considerable problems when mask ventilation or endotracheal intubation is attempted. In this case report the difficulties in anesthetic management of patients with micrognathia and repeated anesthetic courses of a child with Stickler syndrome are presented. It is vital to detect the syndrome in early stages so that adequate counseling and treatment may be given to avoid the potentially irreversible and disabling consequences. PMID- 17138164 TI - Ramsay-Hunt syndrome with vesicular stomatitis in a 4-year-old infant. AB - Ramsay-Hunt syndrome (RHS) usually affects adults, but rare cases of preschool children with RHS have been reported. We report a case of RHS in a healthy 4-year old girl. At the age of 4 years and 5 months, she complained of pain in her mouth and herpes zoster vesicles were noted on the left soft palate and tongue without left pinna, and complete left facial paralysis subsequently developed. She was treated with acyclovir and steroids. Six months later, her facial paralysis had almost fully resolved. PMID- 17138166 TI - The treatment of unusual foreign objects in a root canal: a case report. AB - A 14-year-old female with a history of unsuccessful endodontic treatment of a maxillary central incisor was referred for treatment. Radiographic examination showed an unusual foreign object in the root canal. This report describes the permanent successful treatment of the tooth. PMID- 17138165 TI - Evaluation of the formocresol versus mineral trioxide aggregate primary molar pulpotomy: a meta-analysis. AB - OBJECTIVE: To apply meta-analysis to compare the clinical and radiographic effects of mineral trioxide aggregate (MTA) with formocresol (FC) when used as wound dressing for pulpotomy of primary molars. STUDY DESIGN: The study list was obtained by searching MEDLINE, The Cochrane Library, EMBASE, and SCI. Only those papers that met the inclusion criteria were analyzed. RESULTS: Six studies met the inclusion criteria. There was significant difference between the success rates of FC- and MTA-treated pulpotomized primary molars (P < .05). Clinical assessments and radiographic findings of the MTA versus FC pulpotomy suggested that MTA was superior to FC in pulpotomy resulting in a lower failure rate, with the RR (Relative Risk) being 0.32 (95% confidence interval [CI] 0.11 to 0.90) and 0.31 (95% CI 0.13 to 0.74), respectively. Internal root resorption happened less in the MTA group with RR 0.29, 95% CI 0.11 to 0.77. CONCLUSION: MTA induces less undesirable responses and might be FC's suitable replacement. PMID- 17138167 TI - Biocompatibility of furcal perforation repair material using cell culture technique: Ketac Molar versus ProRoot MTA. AB - OBJECTIVE: The objective of this study was to evaluate the cytotoxicity of furcal perforation repair materials, GI and MTA, using cell culture technique. STUDY DESIGN: The extract of ProRoot MTA and Ketac Molar were treated on PDL cells in a 96-well tissue-culture plate. Cell proliferation after an incubation period of 3 days was determined by using MTT assay. RESULTS: The growth of cultured human periodontal fibroblast cells were suppressed by both perforation repair materials. The percent of cell viability in the Ketac Molar group was lower than in the ProRoot MTA group (P = .000). CONCLUSIONS: Although Ketac Molar has the advantage of adhering to dentine, it is more cytotoxic to the PDL cells than MTA. In selecting the perforation repair material, it is recommended not only to consider the sealing ability of the material with dentine but also the biocompatibility of material to the underlying tissue. PMID- 17138168 TI - Treatment of external inflammatory root resorption after autogenous tooth transplantation: case report. AB - This paper describes a case of autogenous tooth transplantation. An external inflammatory root resorption developed in a 30-year-old female patient and was successfully treated with the placement of calcium hydroxide-based intracanal dressings. Autotransplantation of tooth 32 into the extraction socket of tooth 30 was carried out with no intraoperative complications and uneventful postoperative course. However, the patient did not appear on the 14th postoperative day to initiate endodontic therapy of the transplanted tooth. She did not return for 3 months, at which time the clinical and periapical radiographic examinations revealed the presence of external inflammatory root resorption. Endodontic treatment was initiated immediately. Root canals were prepared and filled with a calcium hydroxide-based intracanal dressing (Calen paste), changed every 21 days during a 6-month period, and were thereafter obturated with Sealapex calcium hydroxide-based sealer and gutta-percha points. After 2 years of follow-up, the transplanted tooth was symptom free with no exacerbated sensibility to percussion, normal mobility, occlusion, and masticatory function. Probing revealed no periodontal pockets or other pathological signs. Radiographic examination showed the periradicular area with normal appearance, completely healed resorption areas, and presence of an intact lamina dura. PMID- 17138170 TI - Radiopaque lesion with peripheral radiolucency of the mandible. PMID- 17138171 TI - A new evaluation method for neurosensory disturbance in the chin of patients undergoing mandibular sagittal split ramus osteotomy: an application of the heat flux technique. AB - OBJECTIVE: To examine neurosensory changes in the chin following bilateral sagittal split ramus osteotomy (BSSRO) using the heat flux technique. STUDY DESIGN: Twelve healthy adults (C-group) and 23 patients undergoing BSSRO (P group) were examined. The warm sense threshold (WHF) and cold sense threshold (CHF) in the chin of these patients were measured by the heat flux technique. In the P-group, touch sensation also was measured with a visual analog scale (VAS) and the Semmes-Weinstein monofilament test (SW test). RESULTS: Both WHF and CHF were significantly larger in the P-group than in the C-group (P = .024). Touch sensation (VAS and SW test) were damaged in some patients of the P-group. However, no correlation was revealed between thermal sensation (WHF and CHF) and touch sensation (VAS and SW test) in the P-group. CONCLUSIONS: These results suggest that the thermal sensation in the chin was damaged after BSSRO. The heat flux technique appears to be one of the useful methods for the examination of neurosensory disturbance in the chin. PMID- 17138172 TI - Influence of lower third molar position on the incidence of preoperative complications. AB - OBJECTIVE: The present study sought to relate the clinical outcome of mandibular third molar impaction surgery to the orthopanoramic radiographically determined position of the affected teeth based on the classifications of Pell and Gregory, and Winter. STUDY DESIGN: A retrospective study was made of 165 patients undergoing the surgical extraction of lower third molars at School of Dentistry of the University of Barcelona, Barcelona, Spain. The positions of the 259 extracted third molars were documented according to the classifications of Pell and Gregory, and Winter, together with the type of mucosal and bony coverage involved. These tooth position parameters were radiologically assessed using Radio Memory software and were retrospectively correlated to the observed infectious, noninfectious, and neurological postsurgical outcomes. RESULTS: A statistically significant relationship (P < .05) was recorded between the appearance of infectious complications and the position parameters "Pell and Gregory classification," "mucosal coverage," and "bony coverage." A statistically significant relationship (P < .05) was observed between the appearance of noninfectious problems and the parameter "Winter classification." Finally, a significant association (P < .05) was recorded between the position variables and the existence of unwanted neurological changes. CONCLUSIONS: The mandibular position of the impacted third molar may be able to be correlated to the development of complications resulting from impaction removal. Vertical third molars in positions IIA and IIB of the Pell and Gregory classification, with partial mucosal and bony coverage, are the most susceptible to undesired outcomes. PMID- 17138173 TI - Is permanent maxillary tooth removal without palatal injection possible? AB - OBJECTIVE: The aim of the present study was to demonstrate if articaine HCl could provide palatal anesthesia in maxillary tooth removal without the need for a second palatal injection. STUDY DESIGN: Of 53 patients, 23 had bilateral and 30 had unilateral extractions. In the study group 2 mL of 4% articaine/HCl with 1:100,000 epinephrine was injected into the buccal vestibule of the tooth. After 5 min the extraction was performed. 27 subjects were controls and subjected to the same protocol with palatal injection. All patients completed a Faces Pain Scale (FPS) and a 100-mm Visual Analogue Scale (VAS) after extraction. RESULTS: According to VAS and FPS scores, when permanent maxillary tooth removal with palatal injection (97.5%) and without palatal injection (96.8%) were compared the difference was not statistically significant (P > .05) [corrected]. CONCLUSION: Permanent removal of maxillary teeth without palatal injection is possible by depositing 2 mL articaine/HCl to the buccal vestibule of the tooth. PMID- 17138174 TI - Cervical pharyngostomy: an alternative approach to enteral feeding. AB - Surgical patients may be unable or unwilling to feed normally, owing to mechanical obstruction to ingestion because of nausea or anorexia or secondary to neurologic abnormality. Such patients may be in a malnourished state that compromises their chances of successful recovery. The Nasogastric tube (NGT) has been the standard method of postoperative alimentation in head and neck cancer patients. However, prolonged use of an NGT has been associated with many problems. Percutaneous endoscopic gastrotomy (PEG) tubes are generally easier to manage and more esthetically pleasing than NGTs. However, they are associated with their own set of shortcomings. Problems and complications with traditional NGT led us to evaluate feeding tubes placed through a cervical pharyngostomy. We present our experience with this technique in 15 patients in whom it was used for postoperative feeding after oropharyngeal cancer resection and reconstruction. The cervical pharyngostomy technique has proven to be a safe, reliable, cost effective and convenient method for postoperative feeding. PMID- 17138175 TI - A reason for the use of toluidine blue staining in the presurgical management of patients with oral squamous cell carcinomas. AB - Verification of oral cancer relies on histopathological diagnosis of suspect or malignant lesions. There is evidence for further pre-surgical screening procedures to localize tumor borders and define other malignant lesions. Important methods are: visual examination, including pan-endoscopy, fluorescence imaging, and brush biopsy, as well as radiologic techniques such as conventional radiography, computed tomography, magnetic resonance imaging, scintigraphy, and ultrasonography, which may reduce the mortality rate associated with oral cancer. In addition, toluidine blue staining is a simple, inexpensive, and excellent diagnostic tool. Herein we show that the clinical use of in vivo staining is effective to define the superficial tumor borders and to detect malignant or pre malignant cells in the surrounding area of the tumor following detection of a malignancy of the oral cavity. In our reported case, the main tumor mass was surrounded by layers of an intact mucosa, yet in a distance of more than 1 cm a group of malignant or pre-malignant cells in the surrounding area required a resection of the tumor in a size that would have been unaddressed during visual examination alone followed by clinical routine program of presurgical examinations after the detection of a malignancy of the oral cavity. PMID- 17138176 TI - Tacrolimus powder in Orabase 0.1% for the treatment of oral lichen planus and oral lichenoid lesions: an open clinical trial. AB - OBJECTIVES: The aim of our study was to evaluate the clinical efficacy and safety profile of a novel compound, Tacrolimus powder in Orabase 0.1% in patients with LP and LL. STUDY DESIGN: Seven patients with LP and 3 with LL were asked to participate. All patients received a 1 week treatment of Fluconazole, prior to entering the study, and on follow up visit were provided with a 15 g container of the study medication. Patients were asked to treat the most symptomatic site, three times a day for two weeks. RESULTS: Disease control (signs) was achieved in most patients by the end of two weeks (from 1.58 to 0.55); all patients experienced a high degree of discomfort (pain) at baseline, which dropped quickly by the end of the second week of treatment (from 1.95 to 0.45); none developed yeast during the course of treatment. Recurrent headaches were reported by one patient with erosive LP, and transient burning by a patient with reticular LP. CONCLUSIONS: Tacrolimus powder in Orabase 0.1% appears to have a relatively safe profile, and represents a likely alternative to topical steroids in the treatment of LP and LL, especially in those at risk for oral candidiasis. PMID- 17138177 TI - Brain activity during stimulation of the trigeminal nerve with noxious heat. AB - OBJECTIVE: The aim of this study was to observe areas of brain activation with painful hot stimulation to the trigeminal nerve. STUDY DESIGN: Nine healthy pain free women (mean age 26.2 +/- 6.9 yrs) with a natural, regular menstrual cycle participated in the study. Whole-brain functional magnetic resonance imaging (fMRI) data were acquired for each participant on day 2 or 3 after the onset of menses using echo-planar imaging at 1.5T with near-isotropic spatial resolution and a temporal resolution of 4 s. RESULTS: Whole-brain fMRI with a Peltier thermode inside the head coil yielded a feasible imaging protocol with little disturbance from the thermode. Painful thermal stimulation of the left trigeminal system activated discrete brain regions within the insula, cingulate gyrus, thalamus, inferior parietal lobe/postcentral gyrus, right middle and inferior frontal gyri, cuneus, precuneus, and precentral gyrus. CONCLUSION: Painful stimulation of the trigeminal nerve resulted in activation of similar brain areas generally known for pain processing of painful peripheral stimulation. PMID- 17138178 TI - Adequacy of training in oral cancer prevention and screening as self-assessed by physicians, nurse practitioners, and dental health professionals. AB - OBJECTIVE: To assess the educational preparedness of medical and dental professionals to reduce the burden of OPC through planning effective cancer control strategies such as reducing tobacco consumption, suggesting healthy lifestyle and diet, and performing early detection through screening examinations and appropriate follow-up. METHODS: Self-reported adequacy of training in oral and pharyngeal cancer (OPC) early intervention activities was compared among 4 health care provider groups in North Carolina. Pretested surveys were mailed to random samples of licensed professionals. RESULTS: Nearly all providers agreed that early detection improves 5-year survival rates from OPC. Compared with 567 medical providers (273 family physicians and 294 nurse practitioners), 1235 dental providers (584 dentists and 651 hygienists) were significantly less likely to feel adequately trained in tobacco and alcohol cessation and to palpate neck nodes, but were significantly more likely to feel adequately trained to perform oral cancer examinations. Among dental providers, those who felt they had adequate training in tobacco and alcohol cessation were significantly more likely to assess these risk factors on patient medical histories. CONCLUSIONS: Education is needed to prepare dental providers to undertake OPC prevention activities, whereas medical providers would benefit from enhanced oral examination skills to improve their performance in early detection. PMID- 17138180 TI - Cystic variant of calcifying epithelial odontogenic tumor. AB - Calcifying epithelial odontogenic tumor (CEOT) is a benign, locally aggressive odontogenic neoplasm characterized by sheets and nests of epithelial cells with deeply eosinophilic or occasionally clear cytoplasm, calcifications, and eosinophilic amorphous material that stains positive for amyloid. Although many cases of CEOT are associated with impacted teeth and occasionally appear radiographically as dentigerous cysts, a true cystic variant has not been previously reported. We report a 15-year-old white male with a large cystic maxillary lesion that filled most of the left maxillary sinus. It deformed the medial wall, the inferior orbital floor, and caused narrowing of the left inferior meatus. Histologically, the cystic lining showed characteristics of CEOT. An intraluminal component that featured histologic characteristics of CEOT was identified during surgery. The lesion was enucleated and the postsurgical course of the patient was uneventful. Because follow-up has been for less than 1 year, a meaningful long-term prognosis cannot be determined at present. However, the patient has not reported any symptoms or signs of recurrence during the follow-up period. PMID- 17138179 TI - Clinically aggressive central giant cell granulomas in two patients with neurofibromatosis 1. AB - BACKGROUND: Neurofibromatosis 1 (NF1) is an autosomal dominantly inherited disorder caused by a spectrum of mutations affecting the Nf1 gene. Affected patients develop benign and malignant tumors at an increased frequency. Clinical findings include multiple cutaneous cafe-au-lait pigmentations, neurofibromas, axillary freckling, optic gliomas, benign iris hamartomas (Lisch nodules), scoliosis, and poorly defined soft tissue lesions of the skeleton. Kerl first reported an association of NF1 with multiple central giant cell granulomas (CGCGs) of the jaws. There have since been 4 additional published cases of NF1 patients with CGCGs of the jaws. CLINICAL CASES: We report on 2 patients who presented with NF1 and aggressive CGCGs of the jaws. In both cases, the clinical course was characterized by numerous recurrences despite mechanical curettage and surgical resection. CONCLUSIONS: We review proposed mechanisms to explain the apparent association between NF1 and an increased incidence of CGCGs of the jaws. While the presence of CGCGs of the jaws in patients with NF1 could represent either a coincidental association or a true genetic linkage, we propose that this phenomenon is most likely related to NF1-mediated osseous dysplasia. Compared to normal bone, the Nf1-haploinsufficient bone in a patient with NF1 may be less able to remodel in response to as of yet unidentified stimuli (e.g. excessive mechanical stress and/or vascular fragility), and consequently may be more susceptible to developing CGCG-like lesions. Alternatively, the CGCG in NF1 patients could represent a true neoplasm, resulting from additional, as of yet unidentified, genetic alterations to Nf1-haploinsufficient bone. PMID- 17138181 TI - Coexisting sublingual and submental dermoid cysts in an infant. PMID- 17138182 TI - Experimental evaluation of bone quality measuring speed of sound in cadaver mandibles. AB - OBJECTIVES: To demonstrate in vitro the feasibility of speed of sound (SOS) measurements through the mandible and to investigate the relationships between mandibular SOS, local bone mineral density (BMD), and the ratio between trabecular and cortical thicknesses (Tb.Th/Cort.Th). The long-term goal is to find a safe, simple test for bone quality in sites for dental implant placement. METHODS: Excised human mandibles (N = 23) were used for the measurement of sound transmission with two 1.6 MHz transducers. Three regions of interest (ROIs) were selected in each of the specimens, namely incisor, premolar, and molar regions. To determine short-term precision, 10 measurements (with repositioning between measures) were performed for each ROI. Local BMD and mandibular cross-sectional morphological characteristics were determined by dual x-ray absorptiometry (DXA) and computed tomography (CT). RESULTS: The coefficient of variation for SOS was found to be 1%. SOS measurements at different sites were significantly different. A significant linear relationship was found between SOS and BMD (r2 = 0.62; P < .0001), whereas a nonlinear relationship was found between SOS and Tb.Th/Cort.Th (r2 = 0.53; P < .0001). CONCLUSION: This in vitro study demonstrated the feasibility of SOS measurement through the mandible. Mandibular SOS reflects local BMD and Tb.Th/Cort.Th. In vivo studies are now required to confirm the predictive power of SOS measurement for bone quality assessment and its potential usefulness as a clinical diagnostic technique. PMID- 17138183 TI - Case report: B-cell lymphoma of the maxillary sinus. AB - The radiographic manifestation of malignant lesions of the maxillary sinus on dental radiographs may be nonspecific, making it difficult to differentiate the lesion from disease of odontogenic origin or more benign sinus pathoses. A radiopaque mass in the maxillary sinus, resulting from a malignant neoplasm growing within or extending into the sinus, can be easily confused with the mass of a mucous retention pseudocyst. Similarly, a malignant growth in the early stages of development can produce radiographic patterns in the alveolar process that may resemble inflammation of odontogenic origin. A case of B-cell lymphoma is reported. The lesion involved the maxillary alveolar process and sinus, producing such a radiographic pattern on the panoramic radiograph. Radiographic and clinical features that should be considered in establishing a differential diagnosis of malignant disease are discussed. PMID- 17138184 TI - Effects of imaging system and exposure on accurate detection of the second mesio buccal canal in maxillary molar teeth. AB - OBJECTIVES: The objectives of this study were to determine observer ability to detect the second mesio-buccal canal (MB2) in maxillary molars using analog film, CMOS, and photostimulable phosphor x-ray detectors across a wide range of radiation exposures and to determine the optimal exposures for these modalities accurately detecting presence of the MB2. RESEARCH DESIGN: Using 6 experimental models with 2 maxillary molars in each, images were generated with the 3 detector modalities at nominal x-ray beam angulations of 0 degrees and 10 degrees across a range of radiation exposures. Observers independently assessed presence of the MB2 in images presented in random order. Horizontal root sections were then evaluated to determine true MB2 presence. RESULTS: For single images at optimal exposures, the correct number of mesiobuccal canals could be identified 55% of the time for CMOS (RVG 6000), 44% of the time for analog x-ray film (InSight), and 39% of the time for photostimulable phosphor (DenOptix). Statistically significant differences were observed between the imaging modalities (chi2 = 23.4, crucial value = 5.99, P < or = .05). RVG 6000 CMOS outperformed both DenOptix photostimulable phosphor (z score = -5.5) and InSight analog direct exposure x-ray film (z score = 4). Exposure affected the rate of accuracy for MB2 detection across the exposure range tested for RVG 6000 and for film, but not to any appreciable degree for DenOptix. CONCLUSIONS: The CMOS detector (RVG-6000) performed best for evaluating presence of the MB2. This was the only modality to exceed 50% reliability with optimal exposure when single images were considered. PMID- 17138185 TI - Heme oxygenase-1 mediates cytoprotection against nitric oxide-induced cytotoxicity via the cGMP pathway in human pulp cells. AB - OBJECTIVE: This study examined the effects of exogenous nitric oxide (NO) on human pulp cells and the involvement of cyclic 3',5'-monophosphate (cGMP) in pulpal protection induced by heme oxygenase-1 (HO-1) against NO-induced cytotoxicity. STUDY DESIGN: This study investigated cytotoxicity and HO-1 induction in pulp cells induced by the NO donor S-nitroso-N-acetyl-D,L penicillamine (SNAP), by using Western blotting and a cell viability assay. It also investigated whether HO-1 contributes to the cytoprotective effect against the cytotoxicity caused by NO and the relationship between HO-1 and cGMP in the signaling pathway. RESULTS: S-nitroso-N-acetyl-D,L-penicillamine decreased cell viability, but increased HO-1 expression in a concentration- and time-dependent manner in human pulp cells. NO-induced cytotoxicity was inhibited in the presence of hemin (inducer of HO-1), whereas it was enhanced in the presence of zinc protoporphyrin IX (ZnPP IX, HO-1 inhibitor); therefore, the NO-induced cytotoxicity was correlated with HO-1 expression. Pretreatment with a membrane permeable cGMP analog, 8-bromo-cGMP, restored cell death and enhanced the HO-1 protein expression induced by SNAP. By contrast, 1 mM SNAP inhibited guanylate cyclase in pulp cells pretreated with 1H-[1,2,4]oxadiazole[4,3-alpha]quinoxalin-1 one (ODQ), resulting in marked cytotoxicity. CONCLUSION: These findings of a link between HO-1, regulated via the cGMP system and NO-induced cytotoxicity in human pulp cells, suggest a protective role for HO-1 in pulpal inflammation. PMID- 17138186 TI - Chemical analysis of powder and set forms of Portland cement, gray ProRoot MTA, white ProRoot MTA, and gray MTA-Angelus. AB - OBJECTIVE: To evaluate the chemical composition and crystalline structures of Portland cement, gray ProRoot MTA (gray MTA), white ProRoot MTA (white MTA), and gray MTA-Angelus. STUDY DESIGN: X-ray diffraction analysis was used to identify and characterize crystalline phases, and energy dispersive x-ray spectrometer was used to determine the chemical composition of the test materials. Both powder form and set form were examined. RESULTS: The crystalline structure and chemical composition of gray and white MTA were similar except for the presence of iron in gray MTA. Both were composed mainly of bismuth oxide and calcium silicate oxide. Portland cement was composed mainly of calcium silicate oxide and did not contain bismuth oxide. Gray MTA-Angelus had a lower content of bismuth oxide than ProRoot MTA. There were no noticeable differences in the chemical composition and crystalline structures between the powder and set forms of any of the material tested. CONCLUSION: Portland cement differed from the MTA by the absence of bismuth ions and presence of potassium ions. Gray MTA contained a significant amount of iron when compared with white MTA. In addition, gray MTA-Angelus had a lower content of bismuth oxide than ProRoot MTA. PMID- 17138187 TI - Lessons from spinal midazolam: When misuse of messages from preclinical models exposes patients to unnecessary risks. PMID- 17138188 TI - Eponymity and the age of ultrasound: how should new blocks be introduced into clinical practice? PMID- 17138189 TI - Intra-articular morphine for inflammatory pain. PMID- 17138190 TI - An essential skill. PMID- 17138191 TI - Effects of intrathecal midazolam on postoperative analgesia when added to a bupivacaine-clonidine mixture. AB - BACKGROUND: Previous clinical and experimental studies have shown that a midazolam-clonidine mixture has a synergistic antinociceptive effect. This study evaluated the postoperative analgesic effect of adding midazolam to an intrathecal bupivacaine-clonidine mixture. METHODS: One hundred ten patients scheduled to undergo elective lower-extremity surgery were enrolled in this double-blind, randomized trial. Spinal anesthesia was administered by using 1 of 2 mixtures. Group B-C received 12.5 mg isobaric 0.5% bupivacaine, 30 mug clonidine, and 0.4 mL 0.9% saline. Group B-C-M received the B-C mixture plus 2 mg of midazolam in a 0.4-mL solution. Motor and sensory block levels were assessed before, during, and after the procedure until regression of the block to S2. Sedation levels were determined before anesthesia, during surgery, and at the end of the procedure. Postoperative analgesia was assessed every 15 minutes by using a visual analog scale. Duration of sensory and motor blocks was determined based on a modified Bromage scale, and time of the first pain relief request was noted. RESULTS: Duration of sensory block, time of first postoperative analgesic request, and amount of postoperative morphine administered were comparable between groups. However, the motor blockade lasted significantly longer in the B C-M group compared with the B-C group (287 +/- 73 minutes vs 257 +/- 72 minutes, respectively; P < .05). CONCLUSION: Addition of midazolam to an intrathecal B-C mixture does not potentiate postoperative analgesia but prolongs the motor blockade. PMID- 17138192 TI - Intra-articular morphine 5 mg after knee arthroscopy does not produce significant pain relief when administered to patients with moderate to severe pain via an intra-articular catheter. AB - BACKGROUND AND OBJECTIVES: Intra-articular (IA) morphine for postoperative analgesia after knee arthroscopy is controversial. The IA catheter technique for test drug administration allows baseline pain assessment before inclusion. Results from one such randomized controlled trial (RCT) in patients with moderate to severe pain have shown equal effects of IA saline with or without morphine 2 mg. However, the IA catheter technique may have an unintended placebo effect. The aims of this placebo-controlled RCT were (1) to compare the analgesic effect of IA saline 1 mL (placebo) with morphine 5 mg given through an IA catheter and (2) to analyze the impact on pain of immediate or delayed removal of the IA catheter. METHODS: Sixty patients operated under general anesthesia had an IA catheter inserted at the end of arthroscopy. Patients who reported moderate or severe postoperative pain during the following hour were randomized to IA saline 1 mL (placebo) or IA morphine 5 mg and to immediate or delayed removal of IA catheter. RESULTS: Forty of 60 patients (67%) developed moderate to severe pain within 1 hour. In addition, 5 patients experienced intolerable pain and were excluded. Significantly more women (24/26) than men (26/39) reported at least moderate pain (P = .018) during the first hour after surgery. There were no differences between IA morphine 5 mg and placebo in pain intensity or pain relief at any time during the 48-hour observation period. There was no detectable effect on pain intensity of early compared with late removal of the IA catheter. CONCLUSIONS: IA morphine 5 mg does not produce clinically significant pain relief in patients with moderate or severe pain after knee arthroscopy. PMID- 17138193 TI - A comparison of injection at the ulnar and the radial nerve in axillary block using triple stimulation. AB - BACKGROUND AND OBJECTIVES: A triple-stimulation technique for axillary block consists of the localization and injection of 2 nerves, median and musculocutaneous, which lie superior to the axillary artery, and of 1 nerve, the radial, which lies inferior. However, in some patients, the ulnar nerve is located first during the search for the radial nerve. The aim of this study was to verify if an ulnar motor response could be considered a satisfactory endpoint as a radial motor response. METHODS: This study was a prospective, randomized, double-blinded study. Ninety patients received a triple-injection axillary brachial plexus block in which the radial nerve (group RAD) or the ulnar nerve (group ULN) was located and injected inferior to the axillary artery. Patients were assessed for sensory and motor block by a blinded investigator at 5-minute intervals over 30 minutes. RESULTS: A statistically significant higher overall block success rate was recorded in group RAD (91% vs. 73%), and this result was related to a larger success rate for anesthetizing the radial nerve (95% vs. 77%). A statistically significant shorter onset time of sensory block for the radial nerve was recorded in group RAD versus group ULN (9 +/- 5 min vs. 16 +/- 7 min), whereas the reverse was true for the ulnar nerve (13 +/- 7 min for group RAD vs. 10 +/- 3 min for group ULN). The time to perform the block was slightly but statistically significantly shorter in group ULN (6.5 +/- 1.7 min vs. 7.8 +/- 1.8 min). CONCLUSIONS: Local anesthetic injection at the ulnar nerve significantly reduces the efficacy and prolongs the onset time of the radial nerve block when triple-stimulation axillary block is performed. PMID- 17138194 TI - Increase in skin temperature after spinal anesthesia in infants. AB - BACKGROUND AND OBJECTIVES: The relatively stable hemodynamics during spinal anesthesia in infants have been attributed to a less active sympathetic nervous system in comparison with adults. Thus, the authors evaluated sympathetic block primarily by measurement of skin temperature and secondarily by determination of noninvasive blood pressure as an indirect sign of sympatholysis. METHODS: In 15 infants (postconceptual age: 45.0 +/- 4.8 weeks; weight: 4.0 +/- 1.2 kg) scheduled for repair of inguinal hernia under spinal anesthesia, skin temperature at the T4 level and at the plantar foot was measured before and after spinal anesthesia. Spinal anesthesia was induced at the L4/L5 interspace with 0.5% hyperbaric bupivacaine 1 mg/kg with 10 microg/kg adrenaline added. RESULTS: Temperature at the plantar foot after spinal anesthesia rose significantly from 33.0 degrees C +/- 1.3 degrees C to 34.7 degrees C +/- 1.4 degrees C within 10 minutes and to 35.6 degrees C +/- 0.9 degrees C after 20 minutes (P < .0001), whereas the temperature at the thorax remained constant at 35 degrees C to 36 degrees C. Systolic and diastolic blood pressure decreased by 15.9 +/- 11.4 mm Hg and 9.0 +/- 9.2 mm Hg, respectively (P < .01), but remained within normal range in all cases. CONCLUSIONS: The authors found a significant increase in skin temperature of the feet within 10 minutes as a sign of sympatholysis, whereas trunk temperature remained constant. Blood pressure decreased but remained within the normal range, despite the observed sympatholysis. PMID- 17138195 TI - Influence of needle position on lumbar segmental nerve root block selectivity. AB - BACKGROUND AND OBJECTIVES: In patients with chronic low back pain radiating to the leg, segmental nerve root blocks (SNRBs) are performed to predict surgical outcome and identify the putative symptomatic spinal nerve. Epidural spread may lead to false interpretation, affecting clinical decision making. Systematic fluoroscopic analysis of epidural local anesthetic spread and its relationship to needle tip location has not been published to date. Study aims include assessment of epidural local anesthetic spread and its relationship to needle position during fluoroscopy-assisted blocks. METHODS: Patients scheduled for L4, L5, and S1 blocks were included in this prospective observational study. Under fluoroscopy and electrostimulation, they received 0.5 mL of a mixture containing lidocaine 5 mg and iohexol 75 mg. X-rays with needle tip and contrast were scored for no epidural spread (grade 0), local spread epidurally (grade 1), or to adjacent nerve roots (grade 2). RESULTS: Sixty-five patients were analyzed for epidural spread, 62 for needle position. Grade 1 epidural spread occurred in 47% of L4 and 28% of L5 blocks and grade 2 spread in 3 blocks (5%; L5 n = 1, S1 n = 2). For lumbar blocks, the needle was most frequently found in the lateral upper half of the intervertebral foramen. Epidural spread occurred more frequently with medial needle positions (P = .06). CONCLUSION: The findings suggest (P = .06) that the risk of grade 1 and 2 lumbar epidural spread, which results in decreased SNRB selectivity, is greater with medial needle positions in the intervertebral foramen. The variability in anatomic position of the dorsal root ganglion necessitates electrostimulation to guide SNRB in addition to fluoroscopy. PMID- 17138196 TI - Postoperative analgesia and recovery course after major colorectal surgery in elderly patients: a randomized comparison between intrathecal morphine and intravenous PCA morphine. AB - BACKGROUND AND OBJECTIVES: Intrathecal morphine is a widely used method for postoperative pain relief after major abdominal surgery. The aim of this randomized, double-blinded study was to compare intrathecal morphine and intravenous PCA morphine for postoperative analgesia and recovery course after major colorectal surgery in elderly patients. METHODS: After written informed consent, patients >70 years of age were prospectively and randomly assigned to receive either preoperative intrathecal morphine (0.3 mg) and postoperative patient-controlled (PCA) intravenous morphine (IT morphine) or PCA alone (group control). Results are presented as mean +/- SD (95% confidence interval). RESULTS: Twenty-six patients successfully completed the study in each group. In the IT morphine group, rate of awakening was delayed. Pain intensity and daily intravenous morphine consumption were significantly reduced 1 and 2 days after surgery in the IT morphine group (P < .01). Mental function (assessed by Mini Mental State and Digit Symbol Substitution Test) was similar in both groups. Episodes of postoperative delirium/confusion occurred similarly in both groups. Time to ileus resolution and time to ambulation without assistance did not differ between the 2 groups. The duration of hospitalization was 8.4 +/- 1.7 (7-11) days and 7.9 +/- 2.0 (6-9.9) days for control and IT morphine, respectively (nonstatistical difference). Patients in the IT morphine group had longer time to awakening from anesthesia and experienced more sedation. CONCLUSIONS: Intrathecal morphine, as compared with intravenous PCA morphine alone, improves immediate postoperative pain and reduces parenteral morphine consumption but does not improve postoperative recovery in elderly patients after major colorectal surgery. PMID- 17138197 TI - Detection of epithelial cell transfer in spinal areas by light microscopy and determining any tissue coring via cell culture during combined spinal-epidural interventions. AB - BACKGROUND AND OBJECTIVES: Epithelial tissue coring by spinal needles during subarachnoid injections may cause intraspinal epidermal tumors. Previous studies have investigated tissue transfer with different needle types during subarachnoid or epidural injection. This study deals with the transfer of epithelial tissue during combined spinal-epidural (CSE) anesthesia. METHODS: We studied 68 American Society of Anesthesiologists I to III adult patients. CSE anesthesia was induced under aseptic conditions at the L2-3 or L3-4 interspace with patients in the lateral decubitus position. Cerebral spinal fluid, spinal needle stylet, fluid used to flush the interior of the spinal needle, fluid used to wash the exterior of the spinal needle, fluid used to flush the interior of the epidural needle, and fluid used to wash the exterior tip of the epidural needle were examined under light microscopy (n = 30 patients) or incubated in a cell-culture medium (n = 38 patients). Samples were incubated in cell-culture medium alone (n = 13) or in a cell-culture medium for 3 weeks and then in a medium with epidermal growth factor (n = 25). As a positive control, skin tissue samples were taken by punch biopsy from 10 randomly chosen patients who underwent CSE interventions. These samples were incubated in an enriched medium serum. RESULTS: Light microscopy revealed that there was cell transfer in all phases in various rates: samples 1, 2, 3, 4, 5, and 6 contained epithelial cells and debris in ratios of 6.9%, 20.7%, 6.9%, 20.7%, 26.7%, and 33.3%, respectively. Epithelial cell colonization was detected in the cell-culture samples taken from the control group but not in the samples taken from the CSE group. CONCLUSIONS: We could not reproduce the cells or cell debris obtained during the CSE interventions in vivo, which can be explained by a possible structural deformation of cells or the inadequacy of the amount of cells that were transferred. PMID- 17138198 TI - Fentanyl iontophoretic transdermal system for acute-pain management after orthopedic surgery: a comparative study with morphine intravenous patient controlled analgesia. AB - BACKGROUND AND OBJECTIVES: The fentanyl HCl iontophoretic transdermal system (ITS) has been demonstrated in clinical trials to be safe and effective for acute pain management after several types of major surgery. The current study compared the efficacy, safety, and convenience of fentanyl ITS with morphine intravenous patient-controlled analgesia (IV PCA) for acute-pain management after unilateral total-hip replacement (THR). METHODS: In this multicenter (52 sites), randomized, open-label, active-controlled, phase IIIb study, patients (n = 799) received fentanyl ITS (40 mug fentanyl [10-minute infusion/lockout], up to 6 doses/h) or morphine IV PCA (1-mg morphine bolus [5-minute lockout], up to 10 mg/h) after unilateral THR. The primary efficacy measure was success ratings ("excellent" or "good") on the patient global assessment (PGA) of the method of pain control in the first 24 hours. Pain intensity and adverse events were also assessed. RESULTS: The PGA success ratings (83.0% v 82.2%; difference = 0.9%; 95% CI: -4.4% to 6.1%) and the mean last pain-intensity scores (3.0 v 3.0; difference = 0.0; 95% CI: -0.33 to 0.33) in the first 24 hours were statistically equivalent between fentanyl ITS and morphine IV PCA groups, respectively. The incidence of adverse events was similar between the groups. CONCLUSIONS: Results of this study demonstrate fentanyl ITS and a standard regimen of morphine IV PCA were comparable methods of pain control for management of acute postoperative pain after THR, on the basis of the PGA success ratings and pain intensity in the first 24 hours of treatment. PMID- 17138199 TI - On the edge of the ultrasound screen: Regional anesthesiologists diagnosing nonneural pathology. PMID- 17138200 TI - The middle interscalene block: cadaver study and clinical assessment. AB - BACKGROUND AND OBJECTIVES: A variety of brachial plexus block techniques via the interscalene approach have been proposed. We describe here a new middle interscalene perivascular approach to the brachial plexus. To verify its effectiveness, we studied 719 patients scheduled for shoulder arthroscopy. Furthermore, to verify the accuracy of the proposed bony landmarks to use in the case of inability to palpate the subclavian artery pulse, we simulated the block on 10 cadavers. METHODS: The aim of our technique is to cannulate the neurovascular bundle by inserting a 35-mm needle lateral to the subclavian arterial pulse near the midpoint of the upper edge of the clavicle in a horizontal or slightly cephalad direction while pointing toward the seventh cervical vertebra. If the pulse of the subclavian artery is not palpable, we localize the direction of the needle with reference to 3 bony landmarks (the middle point of the clavicle, the spinous process of C7, and the sternoclavicular joint). By connecting these 3 landmarks, we obtain an angle whose apex lies at the midpoint of the clavicle and its bisecting line points to the plexus. The needle is introduced in the transverse plane of C7. RESULTS: The block was performed successfully in 692 of 719 cases (96.2%). Horner's syndrome occurred in 93.5% of the cases, arterial puncture with hematoma occurred in <1%, phrenic nerve block without respiratory impairment in 60%, with transient respiratory failure in <1%, and laryngeal nerve block in <1%. The incidence of severe complications or permanent injuries was zero (upper limit 95% confidence interval = 0.4% or 1:250 patients). The technique performed on cadavers showed that the previously mentioned bony landmarks were reliable reference points in reaching the brachial plexus. CONCLUSIONS: Our technique via a middle interscalene approach is easy to perform and provides a high success rate. Even in the absence of a subclavian artery pulse, the easily recognizable bony landmarks reliably guide us in the insertion of the needle. Furthermore, this technique appears to avoid complications that are theoretically possible in other supraclavicular perivascular approaches (pneumothorax) and paravertebral approaches (injection into the vertebral artery and subarachnoidal injection). However, further comparative studies will be required to assess the clinical relevance of the block. PMID- 17138201 TI - Diary of a dream: the American Society of Regional Anesthesia (for Surgery, Obstetrics, and Pain Control). PMID- 17138202 TI - Regional anesthesia and pain medicine after 30 years: A historical perspective. AB - Regional Anesthesia and Pain Medicine celebrates its 30th anniversary in 2006. What began as a 10-page, industry-sponsored bulletin has evolved into a major anesthesiology and pain journal. This history article chronicles the journal's growth and development over 3 decades. PMID- 17138203 TI - Pneumoretroperitoneum as a consequence of fascia iliaca block. PMID- 17138204 TI - Block of the ganglion impar using a coccygeal joint approach. PMID- 17138205 TI - Thoracic paravertebral block: is single-injection really safer? PMID- 17138207 TI - A practical method for removal of entrapped epidural catheters. PMID- 17138208 TI - Needle puncture site and a "walkdown" approach for short-axis alignment during ultrasound-guided blocks. PMID- 17138209 TI - "Credentials" in ultrasound-guided regional blocks. PMID- 17138210 TI - Hydrodynamics of the spinal epidural space. PMID- 17138211 TI - Detection of a retained epidural catheter fragment. PMID- 17138213 TI - Perioperative analgesia with posterior continuous lumbar plexus block for simultaneous ipsilateral total hip and knee arthroplasty. PMID- 17138214 TI - Ultrasound-guided supraclavicular brachial plexus block: a modified Plumb-Bob technique. PMID- 17138216 TI - Campaigning for transparency. PMID- 17138215 TI - Image structure clustering for image quality verification of color retina images in diabetic retinopathy screening. AB - Reliable verification of image quality of retinal screening images is a prerequisite for the development of automatic screening systems for diabetic retinopathy. A system is presented that can automatically determine whether the quality of a retinal screening image is sufficient for automatic analysis. The system is based on the assumption that an image of sufficient quality should contain particular image structures according to a certain pre-defined distribution. We cluster filterbank response vectors to obtain a compact representation of the image structures found within an image. Using this compact representation together with raw histograms of the R, G, and B color planes, a statistical classifier is trained to distinguish normal from low quality images. The presented system does not require any previous segmentation of the image in contrast with previous work. The system was evaluated on a large, representative set of 1000 images obtained in a screening program. The proposed method, using different feature sets and classifiers, was compared with the ratings of a second human observer. The best system, based on a Support Vector Machine, has performance close to optimal with an area under the ROC curve of 0.9968. PMID- 17138217 TI - Keynote comment: importance of recognising scientific evidence. PMID- 17138218 TI - Effect of sentinel-node biopsy on metastatic development in breast cancer. PMID- 17138219 TI - Sentinel-lymph-node biopsy as a staging procedure in breast cancer: update of a randomised controlled study. AB - BACKGROUND: In women with breast cancer, sentinel-lymph-node biopsy (SLNB) provides information that allows surgeons to avoid axillary-lymph-node dissection (ALND) if the SLN does not have metastasis, and has a favourable effect on quality of life. Results of our previous trial showed that SLNB accurately screens the ALN for metastasis in breast cancers of diameter 2 mm or less. We aimed to update this trial with results from longer follow-up. METHODS: Women with breast tumours of diameter 2 cm or less were randomly assigned after breast conserving surgery either to SLNB and total ALND (ALND group), or to SLNB followed by ALND only if the SLN was involved (SLN group). Analysis was restricted to patients whose tumour characteristics met eligibility criteria after treatment. The main endpoints were the number of axillary metastases in women in the SLN group with negative SLNs, staging power of SLNB, and disease free and overall survival. FINDINGS: Of the 257 patients in the ALND group, 83 (32%) had a positive SLN and 174 (68%) had a negative SLN; eight of those with negative SLNs were found to have false-negative SLNs. Of the 259 patients in the SLN group, 92 (36%) had a positive SLN, and 167 (65%) had a negative SLN. One case of overt clinical axillary metastasis was seen in the follow-up of the 167 women in the SLN group who did not receive ALND (ie, one false-negative). After a median follow-up of 79 months (range 15-97), 34 events associated with breast cancer occurred: 18 in the ALND group, and 16 in the SLN group (log-rank p=0.6). The overall 5-year survival of all patients was 96.4% (95% CI 94.1-98.7) in the ALND group and 98.4% (96.9-100) in the SLN group (log-rank p=0.1). INTERPRETATION: SLNB can allow total ALND to be avoided in patients with negative SLNs, while reducing postoperative morbidity and the costs of hospital stay. The finding that only one overt axillary metastasis occurred during follow-up of patients who did not receive ALND (whereas eight cases were expected) could be explained by various hypotheses, including those from cancer-stem-cell research. PMID- 17138220 TI - Effectiveness of switching from adjuvant tamoxifen to anastrozole in postmenopausal women with hormone-sensitive early-stage breast cancer: a meta analysis. AB - BACKGROUND: For more than 20 years, tamoxifen has been the mainstay of adjuvant endocrine therapy for women with hormone-sensitive early-stage breast cancer. However, not only does tamoxifen have potential side-effects such as an increased risk of endometrial cancer and thromboembolic events, but patients can also develop resistance to the drug. We aimed to investigate whether switching treatment of postmenopausal women with such breast cancer to anastrozole after 2 3 years of tamoxifen would be more effective than continuing on tamoxifen for a total of 5 years. METHODS: We did a meta-analysis of three clinical trials--the Austrian Breast and Colorectal Cancer Study Group (ABCSG 8), Arimidex-Nolvadex (ARNO 95), and the Italian Tamoxifen Anastrozole (ITA) studies--in which postmenopausal women with histologically confirmed, hormone-sensitive early-stage breast cancer were randomised to 1 mg/day anastrozole (n=2009) after 2-3 years of tamoxifen treatment or to continued 20 or 30 mg/day tamoxifen (n=1997). We analysed the data with a stratified Cox proportional hazards model with the covariates of age, tumour size, nodal status, grade, surgery, and chemotherapy. FINDINGS: Patients who switched to anastrozole had fewer disease recurrences (92 vs 159) and deaths (66 vs 90) than did those who remained on tamoxifen, resulting in significant improvements in disease-free survival (hazard ratio 0.59 [95% CI 0.48-0.74]; p<0.0001), event-free survival (0.55 [0.42-0.71]; p<0.0001), distant recurrence-free survival (0.61 [0.45-0.83]; p=0.002), and overall survival (0.71 [0.52-0.98]; p=0.04). INTERPRETATION: Our results show that the clinical benefits in terms of event-free survival seen in individual trials for those patients who switched to anastrozole translate into a benefit in overall survival. These findings confirm that clinicians should consider switching postmenopausal women who have taken adjuvant tamoxifen for 2-3 years to anastrozole. PMID- 17138221 TI - Management of Pancoast tumours. AB - Non-small-cell lung carcinomas of the superior sulcus, frequently termed Pancoast tumours, are some of the most challenging thoracic malignant diseases to treat because of their proximity to vital structures at the thoracic inlet. Originally deemed universally fatal, Pancoast tumours are now amenable to curative treatment because of improvements in combined modality therapy and development of new techniques for resection. This review includes discussion of anatomical considerations, initial assessment, multimodality treatment, and surgical approaches for these cancers. PMID- 17138222 TI - Immunobiology of the sentinel lymph node and its potential role for antitumour immunity. AB - The sentinel lymph node (SLN) is thought to be an important lymphoid organ for protecting against metastasis and is thought to play a crucial part in provoking antitumour immunity. Because SLN biopsy is undertaken for various types of cancers, such as malignant melanoma and breast cancer, SLN mapping has become a standard procedure, thereby eliminating unnecessary lymph-node resection in patients who do not have affected nodes. The immune surveillance activities of the SLN in melanoma and breast cancer are thought to be suppressed, whereas in cancers of gastrointestinal-tract, the presence of T cells in the SLN has not been shown to suppress the host's immune function. Furthermore, cell death after primary systemic chemotherapy for solid tumours can provoke an antigen-specific immunity in the tumour, which affects tumour response to treatment and, therefore, survival in patients. This review discusses the immunobiology of the SLN and potential strategies for activation of antitumour immunity by primary systemic chemotherapy and other modalities, in terms of tumour-size reduction and survival benefit. PMID- 17138223 TI - Diet, exercise, and complementary therapies after primary treatment for cancer. AB - Every year, more than 10 million people are diagnosed with cancer worldwide. In view of the substantial improvements in early detection and treatment, even more patients can expect to be alive 5 years after diagnosis. With improvements in longevity, the late-occurring adverse effects of cancer and its treatment are becoming increasingly apparent. Healthy lifestyle behaviours that encompass regular exercise, weight control, healthy nutrition, and some complementary practices--eg, support groups, imagery--have the potential to greatly reduce cancer-treatment-associated morbidity and mortality in cancer survivors and can enhance quality of life. Here, we aim to review the strength of evidence for recommendations for exercise, weight management, nutritional practices, and related complementary therapies; assess the perceived needs of cancer survivors for health information and how they can access this information; and discuss the resources available to oncology care providers and patients about healthy lifestyle behaviours. Overall, this review provides important information to oncology care providers who counsel their patients on preventive lifestyle practices to maximise health and longevity after a diagnosis of cancer. PMID- 17138224 TI - Establishment of ethical oversight of human research in El Salvador: lessons learned. AB - Human research is necessary for establishing the best approaches to health care, and should be overseen by a functional research ethics committee. However, in most low-income countries these committees are inadequate in providing the guidelines needed to ensure that ethics remain a prominent feature of human research. We developed a model of interaction with key institutions in low-income countries that had an interest in improving the infrastructure of human participant research. As a result, a research ethics committee was established at three hospitals in El Salvador. The skills gained from developing these committees and the interest shown by the Salvadoran government in rigorous ethical oversight of research involving human participants has allowed the creation of a national ethics committee, which serves as an entity to establish norms and guidelines for further development of research ethics committees in El Salvador. This model of interaction can also be used by institutions in high income countries to create and strengthen the capacity for ethics in human research in low-income countries. PMID- 17138225 TI - Cervical spine osteosarcoma treated with carbon-ion radiotherapy. PMID- 17138226 TI - Topical imiquimod: effectiveness in intraepithelial melanoma of oral mucosa. PMID- 17138227 TI - Splenic infarction and bevacizumab. PMID- 17138228 TI - Survival advantage with bariatric surgery: Report from the 10th International Congress on Obesity. PMID- 17138229 TI - Improvements in systemic metabolism, anthropometrics, and left ventricular geometry 3 months after bariatric surgery. AB - BACKGROUND: Several lines of evidence have suggested a link between obesity and heart failure, including chronic inflammation, increased sympathetic tone, and insulin resistance. The goal of this study was to evaluate the changes in systemic metabolism, anthropometrics, and left ventricular (LV) contraction, as well as geometry, in clinically severe obese women after bariatric surgery. METHODS: Enrollment was offered consecutively to 22 women with clinically severe obesity. Participants underwent abdominal magnetic resonance imaging to quantify the visceral adipose tissue (VAT) area and tissue Doppler imaging echocardiography to measure the LV contractile function. Fasting blood chemistries were drawn to measure inflammatory markers and to calculate insulin sensitivity. All tests were performed before surgery and 3 months postoperatively. RESULTS: Three months after surgery, a significant increase in insulin sensitivity (mean change +/- SEM 34.0 +/- 10.4, P < .0001) was present. The VAT area had significantly decreased (-66.1 +/- 17.8 cm2, P = .002) and was associated with decreases in body mass index, serum glucose concentrations, and high-sensitivity C-reactive protein levels (r = .61 and P = .005, r = .48 and P = .033, and r = .53 and P = .016, respectively). The LV mass decreased significantly (-3.8 +/- 1.7 g/m(2.7), P = .037), and this decrease was associated with a decrease in glucose concentration (r = .46, P = .041). The LV systolic and diastolic contractile function were normal at baseline, and no change occurred after surgery. CONCLUSION: The early phase of weight loss after bariatric surgery produces favorable changes in LV geometry, and these are associated with normalization in the glucose metabolism. PMID- 17138231 TI - Laparoscopic bariatric surgery can be safe for treatment of morbid obesity in patients older than 60 years. AB - BACKGROUND: Previous reports have questioned the safety of bariatric surgery in older patients. The aim of this study was to quantify the perioperative morbidity and mortality of older patients undergoing laparoscopic bariatric procedures at our institution. METHODS: A retrospective chart review of all laparoscopic bariatric procedures performed from February 1999 to September 2005 was performed to identify patients at Mount Sinai Medical Center who were older than 60 years at surgery. RESULTS: We identified 55 patients (36 women and 19 men). The mean age was 61.5 years (range 60-70), and the mean body mass index was 46.2 kg/m2 (range 38.1-61.0). Of the 55 patients, 33 (60%) had undergone laparoscopic Roux en-Y gastric bypass, 9 (16%) laparoscopic gastric banding, 7 (13%) laparoscopic biliopancreatic diversion with duodenal switch, 3 (5.5%) laparoscopic revisional surgery, and 3 (5.5%) laparoscopic sleeve gastrectomy. The mean operative time was 2.3 hours (range 1.1-5.5). No patients required conversion to open surgery, and no perioperative mortality occurred within 30 days. The morbidity rate was 7.3% (n = 4). One patient developed an anastomotic bleed that was treated conservatively, and another patient developed an empyema that was successfully drained with a chest tube. That patient also developed a urinary tract infection, and another patient had a wound infection. The mean length of stay was 2.8 days (range 1-14). CONCLUSIONS: In a carefully selected patient population in a medical center with appropriate experience, laparoscopic bariatric surgery can be performed safely with low morbidity and mortality in the elderly population. PMID- 17138232 TI - Correlation of radiographic and endoscopic evaluation of gastrojejunal anastomosis after Roux-en-Y gastric bypass. AB - BACKGROUND: Anastomotic stenosis presents as one of the most common late complications in the postoperative period after bariatric surgery. It is often diagnosed by upper gastrointestinal series (UGIS) and/or upper endoscopy (UE). The aim of this study was to determine whether a correlation exists between the Gastrografin UGIS and UE findings in the determination of gastrojejunal anastomotic strictures after Roux-en-Y gastric bypass (RYGB). METHODS: Between July 2001 and October 2003, all medical records of patients who underwent RYGB at our institution were retrospectively reviewed. The medical records of patients who underwent UE because of symptoms suggestive of gastric outlet obstruction and those of patients who were initially evaluated by Gastrografin UGIS before UE were evaluated further. RESULTS: Of 535 morbidly obese patients who underwent RYGB, 52 (9.7%) had UE and were included in this study. The mean number of UEs performed per patient was 2.67. Of these 52 patients, 30 underwent Gastrografin UGIS before UE. The mean diameter of the anastomosis on the first UE was 5.97 mm and on Gastrografin UGIS was 6.83 mm. A good correlation was found between the Gastrografin UGIS and UE findings using Pearson's correlation coefficient (0.44, P = .02) and single linear regression analysis using the endoscopic diameter as the outcome and radiographic findings as the predictor (beta = 0.27, P = .025, 95% confidence interval 0.30-0.49). CONCLUSION: In our study, the Gastrografin UGIS findings correlated positively with the endoscopic gastrojejunal anastomosis findings in patients with anastomotic stricture who had undergone RYGB. PMID- 17138233 TI - Outcomes after laparoscopic adjustable gastric band repositioning for slippage or pouch dilation. AB - BACKGROUND: Slippage and pouch dilation are the most common surgical complications after laparoscopic adjustable gastric banding, yet few reports have described the specific outcomes after band repositioning for these complications. The objective of this study was to examine the intermediate outcomes of our patients who underwent band repositioning for slippage or pouch dilation. METHODS: From October 2000 to December 2005, 1275 patients underwent laparoscopic adjustable gastric banding at our center. Of these, we retrospectively reviewed the data of the first 40 consecutive patients (92.5% women, mean age 41.7 years, mean preoperative weight 123.3 kg [range 86.2-180.5], mean body mass index 44.8 kg/m2 [range 34.6-66.4]) who presented with anterior slippage (52.5%), posterior slippage (20%), or pouch dilation (27.5%, 7 with associated hiatal hernias) that required band repositioning (95%) or explantation (5%). RESULTS: The average time between laparoscopic adjustable gastric banding and reoperation was 22.9 months. Before band repositioning, the mean weight was 91.5 kg, mean body mass index was 33.2 kg/m(2), and mean percentage of excess weight loss was 49.4% (range 1 79.8%). One patient had a recurrent anterior slippage that required a second band repositioning. Two bands were explanted, one for intraoperative gastric perforation and one at the patient's request. The mean percentage of excess weight loss after band repositioning was 48.1% (range 18.2-77.4%) at an average follow-up of 17.6 months (range 6-36). To date, 38 (95%) of the 40 patients have functioning bands. Co-morbidity resolution was seen in 3 (60%) of 5 patients with diabetes, 13 (65%) of 20 with hypertension, and 8 (72%) of 11 with sleep apnea. CONCLUSION: Laparoscopic band repositioning can result in preservation of most of the initial weight loss and co-morbidity resolution. PMID- 17138235 TI - Effect of gastric bypass surgery on vitamin D nutritional status. AB - BACKGROUND: We previously reported a 60% prevalence of vitamin D (VitD) depletion, defined as a 25-hydroxyvitamin D (25-OHD) level of < or =20 ng/mL, in morbidly obese patients preoperatively. We now report the effect of gastric bypass (GB) on the VitD nutritional status in these patients. METHODS: We prospectively studied 108 morbidly obese patients who had undergone GB. Routine postoperative supplementation consisted of 800 IU VitD and 1500 mg calcium daily. Serum calcium, parathyroid hormone, and 25-OHD were measured before and 1 year after GB. RESULTS: The mean patient age was 46 +/- 9 years, 93% were women, and 72% were white. Preoperatively and at 1 year postoperatively, the prevalence of VitD depletion and hyperparathyroidism (HPT) and the mean 25-OHD level was 53% and 44%, 47% and 39%, and 20 and 24 ng/mL, respectively. One year after GB, the percentage of excess weight loss was 67% and demonstrated significant correlations both positively with 25-OHD and inversely with parathyroid hormone. At both intervals, blacks had a greater incidence of VitD depletion than did whites, and, at 1 year after GB, HPT was more common in patients with VitD depletion (55% versus 26%, P = .002). CONCLUSION: With customary supplementation, VitD nutrition is improved after GB, but VitD depletion persists in almost one half of patients, and blacks are at a significantly greater risk than whites. HPT did not improve, and those with VitD depletion had a significantly greater rate of HPT. Additional prospective studies are needed to determine how to optimize VitD nutrition and avoid potential long-term skeletal complications after GB. PMID- 17138237 TI - Percutaneous computed tomography-guided gastric remnant access after laparoscopic Roux-en-Y gastric bypass. AB - BACKGROUND: The bypassed portion of the stomach is difficult to access and evaluate after Roux-en-Y gastric bypass. Access to the excluded stomach may be needed for nutritional support or decompression owing to acute distension and obstruction. We report our experience with percutaneous, computed tomography (CT) guided gastrostomy tube placement into the gastric remnant after laparoscopic Roux-en-Y gastric bypass (LRYGB). METHODS: Of 569 consecutive LRYGB procedures performed, 9 patients underwent successful percutaneous, CT-guided gastrostomy placement. One additional patient was referred from another facility. We reviewed the indications, interval from surgery to the intervention, interval to removal, complications, and success or outcome of the procedure in our patient population. RESULTS: Ten patients underwent percutaneous, CT-guided gastric remnant gastrostomy tube placement. The indications included distended gastric remnant in 6, nutritional access in 4, and remnant drainage after leak in 1. Of the 10 patients, 2 had undergone previous gastric operations. The attempt at percutaneous gastrostomy was unsuccessful in 1 additional patient, who subsequently required laparoscopic gastrostomy (success rate 91%). CONCLUSION: In selected patients after LRYGB, CT-guided gastrostomy tube placement is safe and efficient. It may be used to manage complications of LRYGB, serve as a bridge to definitive surgery, or offer a convenient route for enteral nutritional support. PMID- 17138238 TI - Reversible neurologic dysfunction caused by severe vitamin deficiency after malabsorptive bariatric surgery. AB - Significant deficiencies of both fat- and water-soluble vitamins have been reported to occur after malabsorptive bariatric surgery. However, despite the potential for neurologic manifestations of such deficiencies, few cases of central neurologic dysfunction have been reported. Our group previously reported reversible neurologic dysfunction as an unusual manifestation of vitamin deficiency in a postjejunoileostomy patient. We report on a second case of reversible neurologic dysfunction associated with severe, prolonged vitamin deficiency many years after jejunoileostomy. Neurologic function returned to normal with vitamin repletion. Patients who have undergone malabsorptive surgery are at risk of late metabolic complications and should be closely monitored. PMID- 17138239 TI - Superior mesenteric artery syndrome after laparoscopic Roux-en-Y gastric bypass. PMID- 17138240 TI - Laparoscopic management of perforated marginal ulcer after laparoscopic Roux-en-Y gastric bypass. PMID- 17138241 TI - The use of a synthetic mesh to provide port site fixation during laparoscopic adjustable gastric banding. PMID- 17138242 TI - A comparison of 4-row versus 6-row staplers in the performance of the gastrojejunostomy during laparoscopic Roux-en-Y gastric bypass. PMID- 17138243 TI - Asherson's syndrome - catastrophic antiphospholipid syndrome. PMID- 17138244 TI - The catastrophic antiphospholipid (Asherson's) syndrome. AB - The catastrophic antiphospholipid syndrome (CAPS, Asherson's syndrome) develops rapidly following an identifiable triggering factor (eg infection, trauma, inadequate coagulation neoplasia, obstetric) in antiphospholipid antibody positive patients. It is most frequently encountered in patients with a primary antiphospholipid syndrome or systemic lupus erythematosus (SLE) or "lupus-like" disease (LLD). It manifests mainly with small vessel thromboses affecting organs (gastrointestinal tract, brain, heart), large vessel occlusions in one-third, manifestations of the systemic inflammatory response syndrome (SIRS), particularly the acute respiratory distress syndrome (ARDS). The mortality is high, although with early and effective therapies, including full parenteral anticoagulation, corticosteroids, plasma exchanges and IV globulins, an improvement in this high death rate has been noted recently. PMID- 17138245 TI - Laboratory studies on pathophysiology of the catastrophic antiphospholipid syndrome. AB - The 'catastrophic' variant of the antiphospholipid syndrome (APS) is characterized by a diffuse thrombotic microvasculopathy. In contrast to the classical APS, single venous or arterial medium-to-large blood vessel occlusions are uncommon. The mechanisms of catastrophic APS are not clearly understood. In addition, there are no studies on pathophysiologic mechanisms of catastrophic APS. The clinical manifestations of catastrophic APS probably depend on (a) the organs affected by the thrombotic events and extent of the thrombosis and (b) manifestations of the systemic inflammatory response syndrome which are presumed to be due to excessive cytokine release from affected and necrotic tissues. The evident relationship between APS and infection may enable us to explain the development of catastrophic APS using the sepsis model. This is because catastrophic APS is characterized by multiple microvascular thrombotic events, of rapid onset, and causing multiorgan failure, a picture suggestive of septic shock, in which, there is a massive, acute inflammatory response. PMID- 17138246 TI - Mortality in the catastrophic antiphospholipid syndrome: causes of death and prognostic factors. AB - In order to know the causes of death and the prognostic factors, our group analyzed 250 patients included until February 2005 in the web-site based international registry of patients with catastrophic antiphospholipid syndrome (APS) ("CAPS Registry") (http://www.med.ub.es/MIMMUN/FORUM/CAPS.HTM). Cerebral involvement, mainly consisting of stroke, followed by cardiac involvement and infections were considered the main causes of death in patients with catastrophic APS. The presence of systemic lupus erythematosus was related with higher mortality. According to the results of this analysis, anticoagulation plus steroids plus plasma exchange should be the first line of therapy in patients with catastrophic APS. PMID- 17138247 TI - New subsets of the antiphospholipid syndrome in 2006: "PRE-APS" (probable APS) and microangiopathic antiphospholipid syndromes ("MAPS"). AB - The concept of "probable" antiphospholipid syndrome (APS) is almost identical with several conditions which may presage the development of the APS with its major complications of large vessel thromboses resulting in deep vein occlusions in the lower limbs (DVT) particularly and strokes. These conditions comprising livedo reticularis, chorea, thrombocytopenia, fetal loss and valve lesions. These conditions, comprising livedo reticularis, chorea, thrombocytopenia, fetal loss and valve lesions may be followed, often years later by diagnosable APS. The issue whether these patients should be more aggressively treated on presentation in order to prevent the thrombotic complications. A new subset of the APS is proposed viz. microangiopathic antiphospholipid syndrome ("MAPS") comprising those patients presenting with thrombotic microangiopathy and demonstrable antiphospholipid antibodies who may share common although not identical provoking factors (e.g. infections, drugs), clinical manifestations and haematological manifestations (severe thrombocytopenia, hemolytic anaemia) and treatments viz. plasma exchange. Patients without large vessel occlusions may be included in the MAPS subset. These conditions include thrombotic thrombocytopenic purpura (TTP), hemolytic-uremic syndrome (HUS), and the HELLP syndrome. Patients with catastrophic antiphospholipid syndrome (CAPS) who do not demonstrate large vessel occlusions also fall into this group. Disseminated intravascular coagulation (DIC) has also been reported with demonstrable antiphospholipid antibodies and also manifests severe thrombocytopenia and small vessel occlusions. It may cause problems in differential diagnosis. PMID- 17138248 TI - Lessons from the catastrophic antiphospholipid syndrome (CAPS) registry. AB - Although less than 1% of patients with the antiphospholipid syndrome (APS) develop the catastrophic variant, its potentially lethal outcome emphasizes its importance in clinical medicine today. However, the rarity of this variant makes it extraordinarily difficult to study in any systematic way. In order to put together all the published case reports as well as the new diagnosed cases from all over the world, an international registry of patients with catastrophic APS ("CAPS Registry") was created in 2000 by the European Forum on Antiphospholipid Antibodies. Currently, it documents the entire clinical, laboratory and therapeutic data of more than 300 patients whose data has been fully registered. This registry can be freely consulted at the Internet (www.med.ub.es/MIMMUN/FORUM/CAPS.HTM) and it is expected that the periodical analysis of these data will allow us to increase our knowledge of this condition. PMID- 17138249 TI - Pregnancy and puerperium are high susceptibility periods for the development of catastrophic antiphospholipid syndrome. AB - It is well known that antiphospholipid syndrome (APS) is associated with recurrent pregnancies losses, but is also associated with other obstetric features such as preeclampsia, uteroplacental insufficiency and preterm birth. Pregnancy is a hypercoagulable state than can be complicated by thrombosis, especially in those patients with an underlying thrombophilic disorder. Catastrophic APS is a rare form of presentation of the APS. Several trigger factors have been related with the catastrophic APS, including infections, anticoagulation withdrawal, surgery, neoplasms and lupus "flares". In around 6% of the cases, the catastrophic APS can appear during pregnancy or puerperium. We review this specific subset of the catastrophic APS and propose a therapeutical approach for this particular situation. PMID- 17138250 TI - Catastrophic antiphospholipid (Asherson's) syndrome and genetic thrombophilic disorders in obstetrics. AB - Catastrophic antiphospholipid syndrome (CAPS) (Asherson's Syndrome), is a life threatening condition characterized by a rapidly progressive thromboses resulting in a multiorgan dysfunction syndrome (MODS), evidence of systemic inflammatory response syndrome (SIRS) in the presence of antiphospholipid antibodies. CAPS differs from the classic APS by predominantly affecting small vessels, involvement of unusual organs, rapid onset of MODS, and the development of acute respiratory distress syndrome (ARDS) in 25% of patients, which is a feature of SIRS. Obstetric-related multiorgan failure may be a feature of a subset of CAPS more frequently than was previously thought. Patients with obstetric complications should be tested for antiphospholipid antibodies and genetic thrombophilia in order to institute early prophylaxis. Low-molecular-weight heparin is the drug of choice for preventing obstetric complications and CAPS due to its anticoagulant and anti-inflammatory properties. PMID- 17138251 TI - The catastrophic antiphospholipid (Asherson's) syndrome and malignancies. AB - The catastrophic antiphospholipid syndrome is characterised by the rapid chronological development of fulminant thrombotic complications that predominantly affect small vessels. It has been reported as frequently occurring in patients with underlying malignancies. We analysed the web site-based international registry of patients with catastrophic APS. The clinical characteristics of patients with CAPS and an underlying malignancy were evaluated. Of the 262 patients included in the CAPS registry, information on associated malignancies was available in 23 (9%) cases. Haematological malignancies were present in 6 (26%) patients. Four of the patients suffered from lung carcinoma (17%), and two patients (9%) from colon carcinoma. In most of the patients (61%), malignancy was the precipitating factor for CAPS. In 4 patients (17%), however, surgical procedures related to the carcinoma were noted as precipitating factors. In one patient CAPS occurred during allogenic stem cell transplantation after diagnosis of acute lymphoblastic leukemia (ALL). Cerebral manifestations were most common and consisted mainly of cerebral infarcts and encephalopathy. Recovery occurred in 9/23 (39%) patients. Malignancy may be an important risk factor for CAPS. 9% of patients with CAPS presented with an underlying malignancy. In most of these patients, the malignancy and/or surgical procedures were the precipitating factors for CAPS. PMID- 17138252 TI - Therapeutic and prognostic considerations in catastrophic antiphospholipid syndrome. AB - Catastrophic antiphospholipid syndrome (CAPS) is a rapidly progressive life threatening disease that causes multiple organ thromboses in the presence of antiphospholipid antibodies. High index of clinical suspicion and careful investigation are required to make an early diagnosis so that treatment with anticoagulation, corticosteroids, and plasma exchange or intravenous immunoglobulins can be initiated. Despite this multi-modal treatment, CAPS is associated with high mortality; evidence-based management recommendations do not exist due to the rarity of the condition and the lack of controlled studies. This article reviews the therapeutic and prognostic controversies that were addressed during the 1st International Symposium on CAPS. PMID- 17138253 TI - IgA antibodies to beta2-glycoprotein I and atherosclerosis. PMID- 17138254 TI - Oncogenic inflammation and autoimmune disease. AB - Many models exist to explain the induction and perpetuation of autoimmune diseases. Despite their validation in a variety of animal models, the basis for autoimmune disease in humans remains unknown. Here, we propose that an important aspect of autoimmune disease is the active participation of the target organ due to endogenously produced co-stimulatory factors that cause prolonged antigen presentation and lymphocyte activation. Evidence suggests that a major source of such endogenous signaling comes from newly transformed cells within the target organ that produce pro-inflammatory factors. PMID- 17138255 TI - Report on the second Asia Autoimmunity Forum 3-5 March 2006, Hong Kong. AB - On March 3-5, 2006, The HK Society of Rheumatology, University of Hong Kong and Singapore National Arthritis Foundation organized the second Asia Autoimmunity Forum (AAF) in Hong Kong which was attended by over 200 delegates from around Asia. More than 20 invited international and regional experts in the field of autoimmune rheumatic diseases spoke on topics including the pathogenetic mechanisms, clinical aspects and novel therapeutic approaches of autoimmune rheumatic diseases. There were 8 plenary lectures and 4 symposia and the AAF provided an excellent avenue to promote the co-ordination and knowledge exchange in the area of autoimmune rheumatic diseases in Asia. PMID- 17138256 TI - Object-centred spatial reference in 4-month-old infants. AB - An appreciation of object-centred spatial relations involves representing a 'within-object' spatial relation across changes in the object orientation. This representational ability is important in adult object recognition [Biederman, I. (1987). Recognition-by-components: A theory of human image understanding. Psychological Review, 94, 115-147; Marr, D., & Nishihara, H. K. (1978). Representation and recognition of the spatial organisation of three-dimensional structure. Proceedings of the Royal Society of London, Series B (Biological Sciences), 200, 269-294; Tarr, M. J., & Pinker, S. (1990). When does human object recognition use a viewer-centred reference frame? Psychological Science, 1, 253 256] and is also thought to be a fundamental component of the mature object concept [Piaget, J. (1954). The Construction of Reality in the Child. Routledge & Kegan-Paul: London, UK. (Originally published in French in 1937)]. An experiment is reported in which eighteen 4-month-old infants were familiarised to a specific spatial relation within an object, across six different orientations of the object. On subsequent test trials the object was presented to the infants in an entirely novel orientation. Between successive test trials the within-object spatial relation was alternated between novel and familiar. The infants demonstrated significant sensitivity of their looking to both the novelty of the stimuli and the order in which novel and familiar stimuli were presented. It is concluded that by 4 months of age infants are able to form object-centred spatial frames of reference. These findings are discussed in the light of our current understanding of the development of object representation during infancy. PMID- 17138257 TI - Three-month-old infants' object recognition across changes in viewpoint using an operant learning procedure. AB - Object knowledge refers to the understanding that all objects share certain properties. Various components of object knowledge (e.g., object occlusion, object causality) have been examined in human infants to determine its developmental origins. Viewpoint invariance--the understanding that an object viewed from different viewpoints is still the same object--is one area of object knowledge, however, that has received less attention. To this end, infants' capacity for viewpoint-invariant perception of multi-part objects was investigated. Three-month-old infants were tested for generalization to an object displayed on a mobile that differed only in orientation (i.e., viewpoint) from a training object. Infants were given experience with a wide range of object views (Experiment 1) or a more restricted range during training (Experiment 2). The results showed that infants generalized between a horizontal and vertical viewpoint (Experiment 1) that they could clearly discriminate between in other contexts (i.e., with restricted view experience, Experiment 2). Overall, the outcome shows that training experience with multiple viewpoints plays an important role in infants' ability to develop a general percept of an object's 3D structure and promotes viewpoint-invariant perception of multi-part objects; in contrast, restricting training experience impedes viewpoint-invariant recognition of multi-part objects. PMID- 17138258 TI - Prenatal, perinatal and neonatal stimulation: a survey of neonatal nurseries. AB - A recent survey was conducted on stimulation of mothers and babies during pregnancy and the neonatal period. The survey was responded to by 82 neonatology staff members from Neonatal Intensive Care Units (NICUs) at hospitals in the United States. Some forms of stimulation were extremely common including (1) skin to-skin following birth in the delivery room (83% of hospitals); (2) containment (swaddling and surrounded by blanket rolls) in the NICU (86%); (3) music in the NICU (72%); (4) rocking in the NICU (85%); (5) kangaroo care (98%); (6) non nutritive sucking during tubefeedings in the NICU (96%); and (7) breastfeeding in the NICU (100%). Other forms of stimulation occurred less frequently including (1) pregnancy massage (19%); (2) labor massage (30%); (3) the Doula (assistant who comforts during labor and delivery) (30%); (4) waterbeds in the NICU (23%); and (5) preterm infant massage in the NICU (38%). PMID- 17138259 TI - The association between maternal smoking and secondhand exposure and autonomic functioning at 2-4 weeks of age. AB - This study examined the association between maternal smoking and exposure to environmental tobacco smoke (ETS) during pregnancy and infant heart rate. Participants were 62 mother-infant dyads where mother smoked during pregnancy, 47 dyads where mother did not smoke during pregnancy but was exposed to ETS and 47 dyads where mother did not smoke and was not exposed to ETS. Average heart rate (HR) and respiratory sinus arrhythmia (RSA) were assessed at 2-4 weeks of age during sleep. Infants in both exposure groups had higher HR and lower RSA than nonexposed infants. Furthermore, boys in both exposure groups had higher HR and lower RSA than girls. These results highlight the importance of considering prenatal ETS exposure and infant gender when assessing infant outcomes. PMID- 17138260 TI - Relational information processing of novel unrelated actions by infants. AB - Declarative memory in infants is often assessed via deferred imitation. Not much is known about the information processing basis of the memory effect found in these experiments. While in the typical deferred imitation study the order of actions remains the same during demonstration and retrieval, in two experiments with n=30 respective n=25, 10- and 11-month-old infants, the order of novel unrelated actions in demonstration and retrieval was varied (same, reversed, mixed). This allowed a separation of item-specific from item-relational information processing. In both experiments best memory performance was found when the order of target actions remained the same during encoding and recall, demonstrating that infants seem to rely on item-specific as well as item relational information which has to be ad hoc constructed while encoding. PMID- 17138261 TI - Newborns of depressed mothers who received moderate versus light pressure massage during pregnancy. AB - Sixty-four neonates (M age=6.8 days) of depressed mothers who received moderate pressure massage versus light pressure massage therapy during pregnancy (month 5 through month 8) were compared on their behaviors during 15-min observations and on their performance on the Brazelton Neonatal Behavior Assessment Scale. The group of neonates whose mothers received moderate pressure massage spent a greater percent of the observation time smiling and vocalizing, and they received better scores on the orientation, motor, excitability, and depression clusters of the Brazelton scale. PMID- 17138262 TI - Infants' responses to arm restraint at 2 and 6 months: a longitudinal study. AB - This study examined the continuity, stability, and change of infants' responses to a frustrating event (i.e., arm restraint) between 2 and 6 months in terms of both negative reactivity and its regulation. Fifty-two healthy, full-term infants and their mothers participated in an arm restraint procedure. Infant behaviors were observed and coded at 3-s intervals. The results showed that infants' reactivity to frustration and their ability to regulate such reactivity significantly changed in level over time. Individual differences in frustration reactivity were stable across the two ages; two regulatory behaviors (i.e., orientation to mother and avoidance) could be observed in the same percentage of babies at both 2 and 6 months. At 6 months, several significant associations between frustration reactivity and infant regulatory behaviors emerged. These findings suggest that the arm restraint procedure may be usefully employed to study individual differences in infants as young as 2 months of age. PMID- 17138263 TI - Maternal soothing and infant stress responses: soothing, crying and adrenocortical activity during inoculation. AB - The relation between maternal soothing and infant stress response during inoculation was examined in a sample of 37 mothers and their 3-month-old infants. The mothers' soothing and the infants' cry vocalizations and the mothers' and the infants' salivary cortisol level pre- and post-injection were analysed. There was a positive relation between infants' cry vocalization post-injection and maternal soothing pre- and post-injection. The sample was divided in two sub-groups depending on whether the mothers evidenced most soothing of the infants in the period before (Preparatory group; n=20) or after (Contingent group; n=17) the syringe injection. In the Preparatory group, the duration of infant cry vocalizations was related to amount of maternal soothing before and after the injection, while cry vocalizations in the Contingent group was related to amount of maternal soothing after the injection. The Contingent infants responded to the injection with a significant increase in cortisol, while there was no increase in the Preparatory infants. The Preparatory infants evidenced significantly longer duration of looking at the target stimuli in a visual marking task, suggesting greater difficulties in disengaging attention. These findings indicate that 3 month-olds' stress responses and their mothers' situational behaviour are mutually regulated. PMID- 17138264 TI - Intervention for infants with brain injury: results of a randomized controlled study. AB - A randomized clinical trail (RCT) employed a 12-month individualized cognitive/sensorimotor stimulation program to look at the efficacy of the intervention on 62 infants with suspected brain injury. The control group infants received the State-funded follow-up program provided by the Los Angeles (LA) Regional Centers while the intervention group received intensive stimulation using the Curriculum and Monitoring System (CAMS) taught by public health nurses (PHNs). The developmental assessments and outcome measures were performed at 6, 12 and 18 months corrected age and included the Bayley motor and mental development, the Home, mother-infant interaction (Nursing Child Assessment Feeding Scale (NCAFS) and Nursing Child Assessment Teaching Scale (NCATS)), parental stress and social support. At 18 months, 43 infants remained in the study. The results indicate that the intervention had minimal positive effects on the Bayley mental and motor development scores of infants in the intervention group. Likewise, the intervention did not contribute to less stress or better mother-infant interaction at 12 or 18 months although there were significant differences in the NCAFS scores favoring the intervention group at 6 months. There was a significant trend, however, for the control group to have a significant decrease over time on the Bayley mental scores. Although the sample was not large and attrition was at 31%, this study provides further support to the minimal effects of stimulation and home intervention for infants with brain injury and who may have more significant factors contributing to their developmental outcome. PMID- 17138265 TI - Memory reactivation in the second year of life. AB - Memory reactivation has not been systematically studied with infants older than 1 year. Currently, three experiments examined the effects of a reactivation treatment (priming) on retention throughout the second postnatal year. Fifteen- and 18-month-olds learned an operant train task, forgot it, received a 2-min or 10-s prime, and later were tested for retention. Although the longer prime was effective for 15-month-olds, 18-month-olds required the shorter prime (Experiment 1). The memory was reactivated after delays two (18 months) and three (15 months) times longer than infants originally remembered it (Experiment 2). The reactivated memory was forgotten as fast as the original memory after the 2-min prime and twice as fast after the 10-s prime (Experiment 3). The fact that reactivation changes quantitatively but not qualitatively throughout infancy suggests that the same mechanism mediates it at all ages. These findings have major implications for the impact of early experience on cognitive development. PMID- 17138266 TI - Associations between the developmental trajectories of visual scanning and disengagement of attention in infants. AB - The relation between the developmental trajectories of visual scanning and disengagement of attention and gaze were examined throughout early infancy. A sample of 10 infants carried out a scanning and a disengagement task with the same visual stimuli six times between 6 and 26 weeks of age. Frequency and latency measures were analyzed using multivariate multilevel models and Monte Carlo analyses. The results suggest that the ability to scan a face or an abstract stimulus evolves slightly earlier than the ability to shift gaze to a newly appeared target in the periphery. This is consistent with the account that the parvocellular stream becomes functional slightly before the magnocellular stream. The study revealed no indications of a positive association between the development of scanning and disengagement on the level of the individual infant. Scanning and disengagement change scores contrasted more with one another than could be expected on the basis of chance. This implies that the magnocellular and the parvocellular stream develop rather independently up to the age of 26 weeks. PMID- 17138267 TI - Exploration or imitation? The effect of music on 4-week-old infants' tongue protrusions. AB - In a newborn imitation paradigm, an auditory stimulus--music--replaced the standard adult behavioral model. Alternating intervals of music and silence affected 4-week-old infants' rates of tongue protruding--evidence that tongue protruding is a general response to interesting distal stimuli. PMID- 17138268 TI - Happy faces are habituated more slowly by infants of depressed mothers. AB - Three-month-old infants of depressed (n=16) and non-depressed mothers (n=16) were habituated to video clips of a female model reciting phrases while posing happy or sad facial/vocal expressions and dishabituated to the alternate expressions. Overall, infants of depressed mothers took longer to habituate the video clips compared to infants of non-depressed mothers, and those assigned to habituate the sad video clips displayed a novelty response or dishabituated to the happy expressions. These findings suggest that 3-month-old infants of depressed mothers discriminate sad from happy expressions, however, they do not appear to perceive sad expressions as novel. PMID- 17138269 TI - Age-related changes in deferred imitation between 6 and 9 months of age. AB - Six- and 9-month-old human infants were tested in a deferred imitation paradigm in which an experimenter performed a single action with a novel object. Although infants of both ages imitated the action when tested immediately, only the 9 month-olds exhibited imitation after a 24-h delay. PMID- 17138270 TI - Development of the 24-h rest-activity pattern in human infants. AB - The development of the 24-h rest-activity pattern was investigated in human infants under naturalistic conditions as assessed by continuous actigraphy. Seven infants and their mothers were recorded for 4 (n=1), 6 (n=5) and 12 months (n=1) after birth. Periodogram analysis of rest-activity data was performed over consecutive 10-day intervals. A weak 24-h rest-activity pattern was already present in some infants during the newborn period. The magnitude of the 24-h component in individual periodograms increased across the first months following a saturating function. The time constants of fitted saturating exponential functions--reflecting the rate of development of the 24-h pattern--ranged from 49 to 110 days (n=6) indicating a large interindividual variability. Furthermore, intraindividual variation was observed; the magnitude of the 24-h rest-activity component showed fluctuations around the trend. Miniaturized actigraphs are ideal tools for long-term longitudinal monitoring of rest-activity behavior in infants. PMID- 17138271 TI - Longitudinal assessment of leg motor activity and sleep patterns in infants with and without Down syndrome. AB - Whether infants with Down syndrome (DS) perform leg movements with the same frequency and quality as their typical development (TD) counterparts is equivocal. Furthermore, the relationship between these early leg movements and later onset of locomotor milestones has only been partially explored. The aims of this study were two-fold: (1) to describe the longitudinal leg activity in infants with and without DS (3-6 months), and (2) to examine sleeping patterns and leg activity during the night. In addition, the relationships between leg activities and sleep patterns with locomotor development were explored. An activity monitor was placed monthly on the infant's ankle for 48 h. Data were analyzed to separate day-night, high-low activity, and sleep fragmentation. The results indicate that infants with DS produced more low intensity activity and more fragmented sleep. These findings are discussed in relation to the influence of early motor activity on achievement of functional motor behavior. PMID- 17138272 TI - Risk factors and stress variables that differentiate depressed from nondepressed pregnant women. AB - Eight hundred ten pregnant women (N=340 depressed and N=470 nondepressed) were recruited at prenatal clinics at around 20 weeks gestational age. The women were diagnosed as depressed based on the Center for Epidemiological Studies Depression scale (CES-D) and the Structured Clinical Interview for Depression (SCID). They were interviewed on several demographic variables, risk factors and stress questionnaires. On average, the depressed pregnant women were younger, had lower education levels and socioeconomic status and were less often married. Fewer of the depressed women and their partners were happy when they were told they were pregnant, a greater number of the depressed women experienced a stressful situation during pregnancy, more of the depressed women were prescribed antibiotics during pregnancy, the depressed women had less optimal obstetric complications scores, and a greater percentage of them delivered prematurely. Finally, the scores of the depressed pregnant women on the stress questionnaires suggested greater depression (CES-D), anxiety (STAI), anger (STAXI), pregnancy anxieties (PAAS) and daily hassles. PMID- 17138273 TI - Early changes in muscle activation patterns of toddlers during walking. AB - Early locomotor behavior has been the focus of considerable attention by developmentalists over several decades. Few studies have addressed explicitly patterns of muscle activity that underlie this coordination pattern. Our purposes were to illustrate a method to determine objectively the onset and offset of muscle firings during early walking and to investigate the emergence of patterns of activation of the core locomotor muscles. We tested eight toddlers as they walked overground at walking onset (max. of 3-6 independent steps) and after three months of walking experience. Surface electrodes monitored activity of the gastrocnemius, tibialis anterior, quadriceps, and hamstrings. We reduced EMG signals to a frame-by-frame designation of "on-off," followed by muscle state and co-contraction analyses, and probability distributions for each muscle's activity across multiple cycles. Our results clearly show that at walking onset muscle activity was highly variable with few, if any, muscles showing recurring patterns of behavior, within or among toddlers. Variability and co-activation decreased with walking experience but remained inconsistent, in contrast to the significant increase in stability shown for joint coordination and endpoint (foot placement) parameters. We propose this trend emerges because of the high number of options (muscle combinations) available. Toddlers learn first to marshal sufficient force to balance and make forward progress but slowly discover how to optimize these resources. PMID- 17138274 TI - Postpartum feeding attitudes, maternal depression, and breastfeeding in Barbados. AB - Maternal feeding attitudes, maternal moods and infant feeding practices during the first 6 months postpartum were assessed in 226 healthy, well-nourished Barbadian mother-infant dyads. Factor analysis of the feeding attitudes questionnaire resulted in six independent factors. The belief that breastfeeding was better than bottle-feeding was associated with higher family income, more information seeking behavior and older maternal age at the time of her first pregnancy. Women who believed that breastfeeding was better at 7 weeks postpartum were also more likely to breastfeed at concurrent and later ages, up to 6 months postpartum. This belief was also associated with less maternal depression at 7 weeks and 6 months. The association between feeding attitudes and actual feeding practices was significant even after correcting for maternal moods and other background variables. Conversely, after controlling for feeding attitudes, maternal mood at 7 weeks was still significantly associated with infant feeding practices at 6 months. Thus, feeding attitudes and maternal moods were closely linked, but each contributed independently and uniquely to different aspects of breastfeeding, especially at 6 months. These findings suggest that early intervention addressing maternal feeding attitudes, may improve the extent of breastfeeding and the health of children in this setting. PMID- 17138275 TI - Anisotropic motion coherence sensitivities to expansion/contraction motion in early infancy. AB - We investigated 2- and 3-month-olds' motion coherence sensitivities to radial expansion/contraction by using the preferential looking method. The infants were tested with a stimulus composed of two dynamic random dot patterns placed side by side: an expansion (or a contraction) pattern and a random directional pattern. The results showed that the 3-month-old infants tested with both a contraction and random directional pattern could discriminate between those two motions significantly, even when the contraction motion coherence was relatively low (50%). On the other hand, the 3-month-old infants who were tested with both expansion and random directional pattern could not discriminate between those two motions. None of the 2-month-old infants showed significant discrimination between the expansion/contraction and random motion patterns. Results of the present study suggest that anisotropic motion coherence sensitivities to radial expansion/contraction emerge at around 3 months of age. PMID- 17138276 TI - The discrimination of facial expressions by typically developing infants and toddlers and those experiencing early institutional care. AB - Early experience likely plays an important role in the development of the ability to discriminate facial expressions of emotion. We posited that compared to children reared with their biological families (n=72), abandoned children being reared in institutions (n=39) should demonstrate impairments in this ability. The visual paired comparison procedure was utilized to assess the abilities of 13- to 30-month-old children to discriminate among multiple pairs of photographs of facial expressions. Both groups exhibited a normative profile of discrimination, with no group differences evident. Such findings suggest that early institutionalization does not affect the ability of 1- to 3-year-olds to discriminate facial expressions of emotion, at least as inferred by the Visual Paired Comparison Procedure. PMID- 17138277 TI - Withdrawn and intrusive maternal interaction style and infant frontal EEG asymmetry shifts in infants of depressed and non-depressed mothers. AB - The effects of maternal interaction style (intrusive/withdrawn) on the development of brain electrical activity were studied in infants of depressed and non-depressed mothers shortly after birth and again at 3-6 months of age. Infants of depressed mothers exhibited significantly greater relative right frontal EEG activation than infants of non-depressed mothers. Infants of depressed withdrawn mothers exhibited greater relative right frontal EEG activation than infants of depressed intrusive mothers. Furthermore, while infants of depressed mothers with intrusive interaction styles showed a shift towards greater relative left frontal EEG activation from birth to 3-6 months, infants of depressed mothers with withdrawn interaction styles showed a shift towards greater relative right frontal EEG activation. PMID- 17138278 TI - Maternal mental health and medical predictors of infant developmental and health problems from pregnancy to one year: does former infertility matter? AB - BACKGROUND: Transition to parenthood involves considerable psychological, social and physiological changes. We examined how prenatal and perinatal mental health and medical conditions predict infant's developmental and health status at 12 months, and whether former infertility and assisted reproduction treatment (ART) affect the predictor model. METHODS: The participants were 520 mothers who filled in questionnaires at the second trimester of pregnancy (T1), and when the child was 2 months (T2) and 12 months old (T3). Depressive and anxiety symptoms indicate reduced levels of mental health, and medical factors involve problems in pregnancy (e.g., high blood pressure and bleeding) and birth complications (pain and loss of blood). Neonatal health refers to, e.g., birth-weight and neurological status. At 12 months mothers reported infants' verbal and motor development and general health status. RESULTS: First, it was shown that generally maternal prenatal anxiety and medical problems in pregnancy together predicted infant's developmental problems at 12 months through poor neonatal health, and medical problems alone also through increased level of birth complications. Second, the predictor models differed according to the history of infertility. In the ART group prenatal depression and anxiety and medical problems together predicted infant's developmental problems through maternal post partum depression, and medical problems also through birth complications, whereas in the spontaneous conception group psychological and medical paths were separated and did not carry on developmental and health problems into 12 months. CONCLUSIONS: Maternal health care should consider both psychological and medical risk paths across the whole transition to parenthood, and be aware of specific mediating paths in the risk groups. PMID- 17138279 TI - Which cues are available to 24-month-olds? Evidence from point-of-gaze measures during search. AB - In previous research 2-year-olds have failed to show knowledge of solidity in a search task in which a ball rolled behind a screen and was stopped by a barrier. The screen had four doors and the barrier was visible above the door hiding the ball. To establish what cues 2-year-olds might be using, precise point-of-gaze measures were taken during the hiding event. A transparent screen with opaque doors provided two cues: (1) the ball could be tracked until it failed to emerge, and (2) the barrier's position could indicate the correct door. Point-of-gaze measures revealed that children failed to use the more indirect cue of the barrier, which requires reasoning and spatial integration. Their search success was predicted only by the more immediate cue of actively tracking the ball. These findings support the claim that children use best those cues directly related to the object's disappearance, while failing to use cues that entail higher cognitive demands. PMID- 17138280 TI - The impact of object size and rigidity on infant reaching. AB - Although the changes in kinematics of infant reaching have been studied, few researchers have investigated the improvement of reaching regarding objects of distinct physical properties. The aim of this longitudinal study was to verify the impact of object size and rigidity on the development of reaching in 4-6 month-old infants. Four infants were observed with a motion capture system during trials with four objects of distinct sizes and rigidity. A total of 188 reaches were analyzed by using the 3D movement reconstruction. Our results showed that reaching frequency, mean velocity, and straightness index increased with age. The number of movement units decreased with age and increased for small objects. Rigidity was not shown to affect reaching trajectories. These findings suggest that infants are capable of perceiving the more relevant object properties, thus using their available motor capabilities to modify the essential variables so that they can reach the target more accurately. PMID- 17138281 TI - Stability of mood states and biochemistry across pregnancy. AB - Four hundred and thirty pregnant women were recruited during their second trimester of pregnancy (M=20 weeks). They were designated depressed (N=172) or nondepressed (N=258) on the Structured Clinical Interview of Depression (SCID) and the Center for Epidemiological Studies Depression scale (CES-D). They were given a second assessment when they were approximately 32 weeks gestational age. At both assessments they were given self-report measures (CES-D, the State Anxiety Inventory, and the State Anger Inventory) and provided urine samples for assays of cortisol, catecholamines (norepinephrine, epinephrine and dopamine) and serotonin. They were also given the VITAS scale for lower back pain and leg pain and a sleep disturbance scale. The stability of mood states and biochemistry across pregnancy (20 and 32 weeks) were assessed inasmuch as mood states, and biochemistry have been noted to predict prematurity and low birthweight. Significant correlations were noted for all variables except serotonin. Relationships between mood states and biochemistry were also noted but only between cortisol and depression, cortisol and anxiety, and epinephrine and anxiety. Significant stability was noted between the 20-week measures and the 32 week measures including depression, anxiety, anger, and cortisol. These were, in turn, correlated with each other and with, low back pain, leg pain, and sleep disturbance. These data suggest the stability of mood states and cortisol across pregnancy. PMID- 17138282 TI - Prenatal cortisol, prematurity and low birthweight. AB - Three hundred depressed pregnant women were recruited at approximately 20 weeks gestation. They were then divided by a median split into high and low urinary cortisol level groups. The high cortisol group had higher CES-D depression scores and higher inhibition (BIS) scores prenatally. Their fetuses had smaller head circumference, abdominal circumference, biparietal diameter and fetal weight. The high cortisol group neonates were shorter gestational age and lower birthweight and they had lower Brazelton habituation and higher Brazelton reflex scores. Discriminant function analyses suggested that cortisol levels more accurately classified short gestation and low birthweight groups than CES-D depression scores. PMID- 17138283 TI - Peer interaction in child care centres at 15 and 23 months: stability and links with children's socio-emotional adjustment. AB - In this longitudinal study, 70 children were observed during 90 min of free play in their child care centres at 15 and 23 months of age. The children (39 boys and 31 girls) attended 51 different care groups in 39 centres. The occurrence and stability of peer interactions in the second year of life and their relations to children's socio-emotional adjustment were examined. The frequency of children's negative initiatives towards peers showed significant inter-individual stability from 15 to 23 months and predicted child aggressive/disruptive behaviour at 23 months as rated by professional caregivers in child care. Involvement in positive interactions with peers, and particularly positive responses to peer behaviours at 15 months, predicted well-being in child care at 23 months as rated by professional caregivers. The frequency of negative initiatives towards peers significantly increased and the frequency of positive responses to peer behaviours significantly decreased from 15 to 23 months. PMID- 17138284 TI - The role of infant soothability in the relation between infant negativity and maternal sensitivity. AB - Maternal perceptions of infant soothability moderated the relation between negative infant temperament and maternal sensitivity. Infant negative temperament and maternal sensitivity were significantly positively related when maternal perception of infant soothability was high and significantly negatively related when maternal perception of infant soothability was low. PMID- 17138285 TI - The still-face response in newborn, 1.5-, and 3-month-old infants. AB - The present study investigated the still-face response to a female stranger in newborn, 1.5-, and 3-month-old infants. The results revealed that 1.5- and 3 month-olds, but not newborns, reliably decreased their visual attention and positive affect when the interaction partner became unresponsive during the still face period. PMID- 17138286 TI - Intention or expression? Four-month-olds' reactions to a sudden still-face. AB - We addressed whether 4-month-old infants are primarily influenced by the expression or intention underlying a sudden still-face response by an adult social partner. Sixteen dyads of 4-month-old infants interacted with an adult who posed a still-face directed at one of the two infants. Infants' gazing and smiling responses confirm that they are primarily influenced by the emotional and contingency loss rather than the intention underlying the adult's still-face. PMID- 17138287 TI - Learning at the breast: preference formation for an artificial scent and its attraction against the odor of maternal milk. AB - Human newborns are known to display spontaneous attraction to the odor of human milk. This study aimed to assess whether the positive response to human milk odor can be explained by nursing-related learning, and whether it can be easily reassigned to a novel odor associated with nursing. Infants were exposed or not to a novel odor (camomile, Ca) during nursing, and tested on day 3-4 for their preference for camomile in comparison with either a scentless control (Exp. 1), a scented control (Exp. 2), or maternal milk (Exp. 3). Prior experience with Ca modified the newborns' responses. While the Ca odor became more attractive than a scented control in the Ca-exposed group, the Ca-non-exposed group did not differentiate either stimulus. In Exp. 3, the Ca-non-exposed group preferred the milk odor to the Ca odor, whereas the Ca-exposed group displayed on average equal attraction to both stimuli. Thus, a novel odor can be learned at the breast, and gain similar attractive power than the odor of mother's milk. In sum, reinforcements related with the early episodes of breastfeeding mediate the rapid development of novel odor preferences in human infants. PMID- 17138288 TI - Frames of reference for anticipatory action in 4-month-old infants. AB - The spatial representations of 4-month-old infants were examined in two experiments using a modified version of the visual expectation paradigm (VExP). The experiments were designed in order to determine what spatial frames of reference were available to infants for making anticipatory saccades. In Experiment 1, we found that infants most often used a retinocentric frame of reference that did not take into account their current eye position in making an anticipatory saccade. However, Experiment 2 revealed that under certain conditions infants are more likely to make anticipatory saccades consistent with a body- or object-centred frame of reference. The main difference between the two experiments was the degree to which the featural properties of the stimuli varied. The results shed light on the development of spatial representations for action in infancy. PMID- 17138289 TI - Disorientation inhibits landmark use in 12-18-month-old infants. AB - Recent research has indicated that, particularly under conditions of inertial disorientation, mammals may be sensitive to landmark configuration geometry at the expense of individual featural information when locating hidden goals. The current study sought to establish whether landmark use could be demonstrated in 12-18-month-old infants with and without a disorientation procedure, and with geometrically ambiguous landmark configurations. A peekaboo paradigm was employed in which infants learned to anticipate a peekaboo event after a cue from two locations within a circular arena, followed by a test trial from a novel position in which no peekaboo occurred after the cue. In all conditions, an isosceles triangle arrangement of landmarks was used, with peekaboo occurring between the landmarks of one of the two equal "sides", thus being geometrically ambiguous. In two conditions, the landmarks were distinctive, and in two further conditions, they were identical. In one of the distinctive conditions and one of the identical landmark conditions, infants underwent a disorientation procedure in between training and test trials. Only oriented infants with distinctive landmarks were successful in test trials, thus suggesting that infants are able to use the individual features of landmarks to locate a goal, but can only do so if oriented. PMID- 17138290 TI - Temperament, distraction, and learning in toddlerhood. AB - The word- and nonword-learning abilities of toddlers were tested under various conditions of environmental distraction, and evaluated with respect to children's temperamental attentional focus. Thirty-nine children and their mothers visited the lab at child age 21-months, where children were exposed to fast-mapping word learning trials and nonlinguistic sequential learning trials. It was found that both word- and nonword-learning were adversely affected by the presentation of environmental distractions. But it was also found that the effect of the distractions sometimes depended on children's level of attentional focus. Specifically, children high in attentional focus were less affected by environmental distractions than children low in attentional focus when attempting to learn from a model, whereas children low in attentional focus demonstrated little learning from the model. Translationally, these results may be of use to child health-care providers investigating possible sources of cognitive and language delay. PMID- 17138291 TI - Toy-oriented changes during early arm movements: hand kinematics. AB - In a recent cross-sectional study, we found that young infants changed their spontaneous arm movements in the presence of a toy, termed 'toy-oriented changes', in systematic ways beginning many weeks before their first consistent reaches [Bhat, A. N., Heathcock, J. H., & Galloway, J. C. (2005). Toy-oriented changes in hand and joint kinematics during the emergence of purposeful reaching. Infant Behavior and Development, 28(4), 445-465]. The purpose of the present study was to test specific hypotheses regarding toy-oriented changes in a longitudinal design. METHODS: Thirteen infants were observed every other week from 8 weeks of age up to the onset of reaching. At each session, hand and joint motions were observed with and without a toy present using a high-speed motion capture system. This paper focuses on the toy-oriented changes in hand variables. RESULTS: As predicted, infants displayed a meaningful pattern of toy-oriented changes, which systematically changed as infants approached the first week of reaching. During the Early phase (8-10 weeks before reaching), infants scaled down their movement length and speed in the presence of a toy. During the Mid phase (4-6 weeks before reaching), infants scaled up movement number and speed, increased movement smoothness, and decreased their hand-toy distance in the presence of a toy. During the Late phase (within 2 weeks of reaching), infants continued to change their hand's position to get closer to the toy and began contacting it. Interestingly, movement number and smoothness displayed similar developmental patterns, where movement length and speed displayed similar patterns. CONCLUSION: Toy-oriented adaptation of arm movements emerges in the first months of life and forms a complex, yet tractable continuum with purposeful reaching. These results provide a foundation to test more specific hypotheses of hand and joint coordination in both typically developing infants and those infants born at risk for coordination impairments. PMID- 17138292 TI - Why don't they just let it go? AB - Discrepancies between what children expect about physical causality (indexed by looking time) and how they act on that knowledge have led to criticisms of claims about what infants "know." Baillargeon [Baillargeon, R. (1999). Young infants' expectations about hidden objects: A reply to three challenges. Developmental Science, 2, 115-163] advocates examining more tasks before revising views of early cognitive development. We report another discrepancy which suggests an additional indicator of what is salient for preverbal infants. While examining the Uzgiris-Hunt test performances of 40 children (26 females), 7.6-26.9-months old, infants appeared captivated by the bouncing of a small rubber ball. However, most infants reproduced the motion of the bounce event itself, repeatedly hitting the ball against the table, rather than the experimenter's action (dropping). Comparing performances of those who did and did not imitate the drop, two possibly interrelated interpretations remained consistent with the data: infants perform goal-directed imitation of interesting phenomena, perhaps because they believe they must apply force to make them happen. PMID- 17138293 TI - Measurement of fine-grained aspects of toddler temperament: the early childhood behavior questionnaire. AB - This article describes the development, reliability, and factor structure of a finely differentiated (18 dimensions) parent-report measure of temperament in 1.5 to 3-year-old children, using a cross-sectional sample (N=317) and a longitudinal sample of primary (N=104) and secondary (N=61) caregivers. Adequate internal consistency was demonstrated for all scales and moderate inter-rater reliability was evident for most scales. Longitudinal stability correlations were primarily large over 6- and 12-month spans and moderate to large from 18 to 36 months. Factor analysis revealed a three-factor structure of Surgency/Extraversion, Negative Affectivity, and Effortful Control. In both samples and for both primary and secondary caregivers, older children received higher scores for Attention Focusing, Discomfort, Inhibitory Control, and Positive Anticipation. Primary caregivers rated females higher in Fear, and lower in High-intensity Pleasure, than males; secondary caregivers rated females higher than males in several aspects of Effortful Control. PMID- 17138294 TI - General to specific development of movement patterns and memory for contingency between actions and events in young infants. AB - To understand infants' learning of the contingency between their actions and events, we studied inter-limb movement patterns of 48 infants aged 2-4 months when they attempted moving a mobile using a string attached to their arm. The session was composed of baseline, acquisition, immediate retention test, re acquisition, interference, and delayed retention test periods. The analysis revealed motor pattern dependence on age--infants exhibited increased movement over base line of all limbs (2-month-olds), both arms (3-month-olds), and the connected arm (4-month-olds). The acquired patterns were produced during immediate and delayed test periods across age groups. The results suggest that 2 month-olds can acquire and retain general body movements that induce contingent changes in a mobile, while 3- and 4-month-olds form memories that serve as a constraint enabling highly specific movement of their arm to effectively activate the mobile. PMID- 17138295 TI - The correlates of dyadic synchrony in high-risk, low-income toddler boys. AB - Synchrony has been broadly conceptualized as the quality of the parent-child dyadic relationship. Parenting, factors that compromise caregiving quality, and child characteristics have all been theoretically linked to synchrony, but little research has been conducted to validate such associations. The present study examined correlates of synchrony including parenting, maternal psychological resources and child attributes, among a sample of 120 mother-son dyads who were participating in a treatment study for children identified as being at risk for developing early conduct problems. There families participated in an at-home assessment, which included a series of mother-son interactions. Synchrony was associated with aspects of parenting and child attributes, including maternal nurturance, and child emotional negativity and language skills. The findings are discussed in terms of parent and child contributions to the development of synchrony. PMID- 17138296 TI - Young infants reach correctly in A-not-B tasks: on the development of stability and perseveration. AB - This paper examines the development of perseverative reaching in the A-not-B task. We describe two recent models that view perseveration as a sign of developmental progress toward stability. In Experiment 1, we test the novel prediction from both models that very young infants should not perseverate in the A-not-B task whereas older infants should. We tracked infants' behavior monthly on the A-not-B task and found that infants reached correctly at 5 months, and only perseverated at 7 and 8 months of age. Experiment 2 provides further evidence on the role of motor development in the emergence of perseveration by exploring the connection between perseveration and detailed changes in reach kinematics in two infants across the first year. These data together suggest that perseveration is a sign of developmental achievement on the path to stable and flexible behavior. PMID- 17138297 TI - Prenatal depression effects on the fetus and newborn: a review. AB - A review of research on prenatal depression effects on the fetus and newborn suggests that they experience prenatal, perinatal and postnatal complications. Fetal activity is elevated, prenatal growth is delayed, and prematurity and low birthweight occur more often. Newborns of depressed mothers then show a biochemical/physiological profile that mimics their mothers' prenatal biochemical/physiological profile including elevated cortisol, lower levels of dopamine and serotonin, greater relative right frontal EEG activation and lower vagal tone. Elevated prenatal maternal cortisol is the strongest predictor of these neonatal outcomes. Moderate pressure massage can alleviate these effects including reducing prematurity. PMID- 17138298 TI - Infant behavioral responses to infant-directed singing and other maternal interactions. AB - Seventy, 6-9-month-old infants were videotaped during six interactions: mother sings assigned song, "stranger" sings assigned song, mother sings song of choice, mother reads book, mother plays with toy, and mother and infant listen to recorded music. Infant-directed (ID) singing conditions elicited moderately positive cognitive behavior, low levels of positive physical behavior and minimal amounts of vocal behaviors, mostly negative. Across all conditions, cognitive scores remained positive at low to moderate levels. Physical responses were most positive during book and toy, most negative during recorded music, and differed by gender, especially during ID singing. Vocally, infants responded positively to toy, and 8-month-old infants vocalized more than younger infants, particularly during ID singing conditions. ID singing appears just as effective as book reading or toy play in sustaining infant attention and far more effective than listening to recorded music, while interactions involving objects may provide opportunity for shared attention. PMID- 17138299 TI - Using "Bubbles" with babies: a new technique for investigating the informational basis of infant perception. AB - The 'Bubbles' technique (Gosselin, F. & Schyns, P.G. (2001). Bubbles: A technique to reveal the use of information in recognition tasks. Vision Research, 41, 2261 2271) has been widely used to reveal the information adults use to make perceptual categorizations. We present, for the first time, an adapted form of Bubbles, suitable for use with young infants. PMID- 17138300 TI - Mothers' depressed mood and anxiety levels are reduced after massaging their preterm infants. AB - Forty mothers whose preterm infants were about to be discharged from the Neonatal Intermediate Care Nursery (NICU) were randomly assigned to two groups: the first group of mothers conducted preterm infant massage and the second group only observed their preterm infants receiving massage. Both groups of mothers had lower depressed mood scores following the session. However, only the group who massaged their infants had lower anxiety scores after the session. PMID- 17138301 TI - Longitudinal expressions of infant's prehension as a function of object properties. AB - This longitudinal study examined the prehensile development of infants (9-37 weeks) under different task constraints (object shape, size and texture). At 9 weeks of age, the infants did not reach or make contact with the objects, but all 10 infants showed goal directed prehensile movement by about 17 weeks. As they continued to age the infants further differentiated an adaptive prehensile grip configuration to the object constraints. The pattern of longitudinal findings provides further evidence that: (1) the constraints of object properties play an important role in channeling the expressions of infant's prehensile functioning and (2) the classic observation by Halverson [Halverson, H. M. (1931). An experimental study of prehension in infants by means of systematic cinema records. Genetic Psychology Monographs, 10, 107-283] of an apparent order to the development of the fundamental prehensile sequence is task dependent. PMID- 17138302 TI - Continuity of temperament from infancy to middle childhood. AB - Continuity of temperament from 6 months (the IBQ) to 5.5 years (the CBQ) was explored in Finnish children (n=231) within the theoretical framework deviced by Rothbart. Activity level, smiling and laughter, distress to limitations and fear showed significant differential homotypic and heterotypic continuity, while soothability and duration of orienting showed significant differential heterotypic continuity. On the level of latent superconstructs, infant positive and negative affectivity accounted for 4.6, 22.3, and 6.0% of the variance in childhood extraversion, effortful control and negative affectivity, respectively. Infant and childhood temperament clustered into profile types named "resilient", "undercontrolled", and "overcontrolled" mirroring ipsative continuity. These findings give empirical credence to Rothbart's theory by replicating and extending previous findings in significant ways. PMID- 17138303 TI - A biomechanical analysis of the 'high guard' position of arms during walking in toddlers. AB - In this study, we investigated biomechanical contributions of the high guard position of the arms observed only in a subgroup of toddlers at very early stages of gait development. Six healthy toddlers who showed this peculiar arm posture were involved in this study. They participated in two data collection sessions (1 month apart). We used three-dimensional analysis of arm posture during gait to estimate the changes in forces and torques generated by the arms and acting on the upper trunk segment. Across visits, toddlers' increase in walking speeds coincided with lowering arm postures. Despite the apparent trend of changes in arm posture in this group of toddlers, the interaction between arm posture and upper trunk position created a variety of changes in forces and torques among individuals. Findings of this study exhibited an example of the exploration of dynamics by toddlers in the early stage of gait development. PMID- 17138304 TI - Instrumental and vocal music effects on EEG and EKG in neonates of depressed and non-depressed mothers. AB - Neonates (M age=16 days) born to depressed and non-depressed mothers were randomly assigned to hear an audiotaped lullaby of instrumental music with vocals or without vocals. Neonatal EEG and EKG were recorded for 2min (baseline) of silence and for 2min of one or the other music presentation. Neonates of non depressed mothers showed greater relative right frontal EEG asymmetry to both types of music, suggesting a withdrawal response. Neonates of depressed mothers on the other hand showed greater relative left frontal EEG asymmetry to the instrumental without vocal segment, suggesting an approach response, and greater relative right frontal EEG asymmetry to the instrumental with vocal segment, suggesting a withdrawal response. Heart rate decelerations occurred following the music onset for both groups of infants, however, compared to infants of non depressed mothers, infants of depressed mothers showed a delayed heart rate deceleration, suggesting slower processing and/or delayed attention. These findings suggest that neonates of depressed and non-depressed mothers show different EKG and EEG responses to instrumental music with versus without vocals. PMID- 17138305 TI - Bed-sharing, breastfeeding and maternal moods in Barbados. AB - Bed-sharing among Barbadian mothers and infants was studied in relationship to maternal and infant characteristics. This prospective study followed 226 healthy, well-nourished mother-infant dyads at birth, 7 weeks, 3 months, and 6 months postpartum. At each age, approximately half of the infants shared the same beds as their mothers. Bed-sharing was associated with demographic characteristics, especially fewer home conveniences, and also maternal characteristics, including less information seeking by the mother and younger maternal age at first pregnancy. Bed-sharing was also associated with lower infant birth weights. Maternal moods were significantly correlated with bed-sharing, such that mothers who reported having more despair and anxiety were also more likely to sleep with their infants. Bed-sharing was also significantly associated with increased breastfeeding at all infant ages, but this relationship was no longer significant once the effects of maternal moods were controlled. This study emphasizes the importance of assessing maternal moods in studies evaluating the risk and benefits of bed-sharing. PMID- 17138306 TI - The role of action effects in 12-month-olds' action control: a comparison of televised model and live model. AB - The present study investigated differences in infant imitation after watching a televised model and a live model and addressed the issue of whether action effects influence infants' action control in both cases. In a 2x2 design, 12 month-old infants observed a live or a televised model performing a three-step action sequence, in which either the 2nd or the 3rd action step was combined with an acoustical action effect. We assumed that infants would use the observed action-effect relations for their own action control in the test phase afterwards. Even though results exhibited differences in the absolute amount of imitation between the two demonstration groups, both groups showed similar result patterns regarding the action effect manipulation: infants imitated the action step that was followed by a salient action effect more often and mostly as the first target action, emphasizing the important role of action effects in infants' action control. PMID- 17138307 TI - Infant recall memory and communication predicts later cognitive development. AB - This longitudinal study investigates the relation between recall memory and communication in infancy and later cognitive development. Twenty-six typically developing Swedish children were tested during infancy for deferred imitation (memory), joint attention (JA), and requesting (nonverbal communication); they also were tested during childhood for language and cognitive competence. Results showed that infants with low performance on both deferred imitation at 9 months and joint attention at 14 months obtained a significantly lower score on a test of cognitive abilities at 4 years of age. This long-term prediction from preverbal infancy to childhood cognition is of interest both to developmental theory and to practice. PMID- 17138308 TI - Joint attention in term and preterm infants at 12 months corrected age: the significance of gender and intervention based on a randomized controlled trial. AB - This study tested the effects of optimized neonatal mother-infant transactions on joint attention performance at 12 months. Surviving infants <2000g from a geographically defined area were randomly assigned to a preterm intervention (n=71) or preterm control group (n=69). Comparisons were made between preterm groups, secondary with a term group (n=75). Joint attention was measured using the Early Social Communication Scales. Preterm intervention infants scored significantly higher than preterm control infants on elements Initiating Joint Attention (p<0.05), Initiating Object Requests (p<0.05), and Responding to Social Interaction (p<0.0005). Intervention was not associated with performance on elements Responding to Joint Attention, or on Responding to Requests. ELBW infants scored significantly lower than VLBW and LBW infants on imperative functions. Girls outperformed boys on all communication elements. An intervention implemented during the neonatal period can be of advantage for certain aspects of joint attention performance in preterm infants. PMID- 17138309 TI - A comparison of between- and within-subjects imitation designs. AB - Two experimental methods, which have dominated the study of declarative memory in preverbal children with imitation tasks, namely the deferred imitation and elicited imitation paradigm, differ in the amount of physical contact with test stimuli afforded infants prior to a test for long-term recall. The current study assessed effects of pre- and post-demonstration contact with test stimuli on deferred imitation of novel, single-step unrelated actions with multiple objects by 8(1/2)- and 10(1/2)-month-old infants (N=50). The rate of target action completion after a delay remained consistent at both ages across different conditions of prior contact with test stimuli. This study shows that a within subjects baseline appraisal is valid within certain experimental parameters and offers a more economical alternative. The results show furthermore that different experimental designs utilized to assess deferred imitation are highly comparable for the first year despite differences in determining baseline. PMID- 17138310 TI - Moderate versus light pressure massage therapy leads to greater weight gain in preterm infants. AB - Sixty-eight preterm infants (M GA=30 weeks) were randomly assigned to a moderate or to a light pressure massage therapy group to receive 15 massages three times per day for 5 days. Behavior state, stress behaviors and heart rate were recorded for 15min before and during the first 15-min therapy session. Weight gain was recorded over the 5-day therapy period. The moderate versus light pressure massage group gained significantly more weight per day. During the behavior observations the moderate versus light pressure massage group showed significantly lower increases from the pre-session to the session recording on: (1) active sleep; (2) fussing; (3) crying; (4) movement; and (5) stress behavior (hiccupping). They also showed a smaller decrease in deep sleep, a greater decrease in heart rate and a greater increase in vagal tone. Thus, the moderate pressure massage therapy group appeared to be more relaxed and less aroused than the light pressure massage group which may have contributed to the greater weight gain of the moderate pressure massage therapy group. PMID- 17138311 TI - Prenatal paternal depression. AB - Prenatal depressive symptoms, anxiety, anger and daily hassles were investigated in 156 depressed and non-depressed pregnant women and their depressed and non depressed partners (fathers-to-be). Depressed versus non-depressed fathers had higher depression, anxiety and daily hassles scores. Although the pregnant women in general had lower anxiety, anger and daily hassles scores than the men, the scores on the measures for depressed fathers and depressed mothers did not differ. Paternal depression appeared to have less effect than maternal depression on their partners' scores. However, the similarity between the scores of depressed mothers and depressed fathers highlights the importance of screening for depression in fathers-to-be as well as mothers-to-be during pregnancy. PMID- 17138312 TI - Preference for peers in infancy. AB - Previous findings suggest that infants prefer other infants from among individuals of various ages. There are two explanations for this: one is that like adults, infants prefer babyish characteristics; the other is that infants prefer individuals who are similar to themselves. We examined whether infants respond differently to same-age infants from among older or younger infants. This might allow the possibility of methodologically separating the two explanations for peer preference in infancy. The results revealed that 9-month-olds showed peer preference when they saw movies of infants. Further experiments showed that 6- and 9-month-olds preferred static images of same-age infants. The lack of preference of 6-month-olds for movies of same-age infants could be related to their ability to process information. Thus, the cue for peer preference may be physical similarity to oneself, rather than the strength of babyish characteristics reflecting a particular developmental stage. PMID- 17138313 TI - Greater arousal and less attentiveness to face/voice stimuli by neonates of depressed mothers on the Brazelton Neonatal Behavioral Assessment Scale. AB - Neonates born to depressed (n=44) versus non-depressed (n=43) mothers were compared on individual items of the Brazelton Neonatal Behavioral Assessment Scale (NBAS). The neonates of depressed mothers received lower scores on orienting to the live face/voice stimulus and on the alertness items, suggesting they were less attentive. They also scored less optimally on the cuddliness and hand-to-mouth activity items, suggesting they were more aroused. These data lend support to the model that infants of depressed mothers are more aroused and less attentive. PMID- 17138314 TI - The effect of encoding time on retention by infants and young children. AB - Using a Visual Recognition Memory (VRM) procedure, we examined the effect of encoding time on retention by 1- and 4-year olds. Irrespective of age, shorter familiarization time reduced retention, and longer familiarization time prolonged retention. The amount of familiarization that yielded retention after a given delay decreased as a function of age. PMID- 17138315 TI - The case of a 52-year-old woman with chronic tooth pain unresolved by multiple traditional dental procedures: an evidence-based review of the diagnosis of trigeminal neuropathic pain. PMID- 17138316 TI - Smoking cessation training improves provider performance but has limited effect on patient smoking behavior. PMID- 17138317 TI - Postoperative antibiotics do not improve clinical outcomes following removal of impacted mandibular third molars. PMID- 17138318 TI - Implant failure is higher in grafted edentulous maxillae. PMID- 17138319 TI - Comparison of 2 treatment concepts for implant-supported bar-retained overdentures demonstrates equivalence regarding efficacy and safety. PMID- 17138320 TI - Conventional mucogingival surgery is more effective than guided tissue regeneration in root coverage procedures. PMID- 17138321 TI - Powered toothbrushes and manual toothbrushes are generally equally effective in plaque removal. PMID- 17138322 TI - Outcome of the root canal treatment on permanent teeth is related to the preoperative diagnosis and the accuracy of the treatment procedure. PMID- 17138323 TI - Smoking may impede gingival bleeding. PMID- 17138324 TI - Moderate coffee intake may reduce the risk of oral, pharyngeal, and esophageal cancer. PMID- 17138326 TI - Healthful eating practices may lower caries prevalence in the primary dentition in children aged 2 to 5 years. PMID- 17138325 TI - Dental anxiety is weakly correlated with oral health-related quality of life. PMID- 17138327 TI - Greater tooth-brushing frequency, fluoride supplement use, and tap water fluoride concentrations above 0.7 mg/L are risk factors for dental fluorosis. PMID- 17138328 TI - Diverse components of the oral environment in attention-deficit hyperactivity disorder (ADHD) make it difficult to establish whether ADHD is a risk factor for dental caries. PMID- 17138329 TI - ALDH2 genotype is associated in the relation between alcohol intake and periodontitis. PMID- 17138330 TI - Human leukocyte antigen combinations may be risk factors for periodontal disease. PMID- 17138331 TI - Helicobacter pylori (HP) may be involved in the pathogenesis of recurrent aphthous ulcers in children and adolescents. PMID- 17138333 TI - Smoking "shisha" (water pipe) or cigarettes may increase the risk for dry socket following extraction of mandibular third molars. PMID- 17138334 TI - Low prevalence of interleukin L-1A(-899) and IL-1B(+3954) genetic polymorphism found in a Thai population group. PMID- 17138335 TI - Interdisciplinary resources optimize Evidence-Based Dental Practice. AB - Evidence-Based Dental Practice (EBDP) is the most appropriate and best patient centered care because it uses the evidence derived from interdisciplinary resources. These resources include the integration of the clinical and basic science researcher, clinician, and patient in the clinical decision-making process. Best evidence has different meanings to each of the interdisciplinary decision makers and, thus, defines their individual roles accordingly. Implementing EBDP requires an organizational structure and process that involve evidence-based dentistry databases in centralized repositories that are shared nationally and internationally. Facilitating the inclusive and reciprocal transfer of information from all resources requires use of information technology and computerized decision-making support programs. However, the potential lack of financial resources and incentives will diminish the universal adoption of EBDP. PMID- 17138336 TI - No evidence that improved personal oral hygiene prevents or controls chronic periodontitis. PMID- 17138337 TI - Children with special health care needs may encounter difficulty finding dental care. PMID- 17138338 TI - The relative contribution of oral cancer risk factors may differ in younger as compared to older adults. PMID- 17138339 TI - A diagnosis of periodontitis was associated with an increased failure rate of dental implants followed for approximately 2.5 years. PMID- 17138340 TI - Oral health-related quality of life is substantially impaired in patients who seek referral to a periodontal specialty practice. PMID- 17138341 TI - Cephalometric norms for Turkish adults with Class I occlusions. PMID- 17138342 TI - Bruxism is significantly more prevalent among smokers. PMID- 17138343 TI - Non-public water use in fluoridated water areas can increase caries risk in primary dentition. PMID- 17138344 TI - Tobacco smoking not found to be related to apical periodontitis. PMID- 17138345 TI - Periodontal disease may cause acute ischemic stroke. PMID- 17138346 TI - Periodontal disease, periapical lesions and caries were, in that order, the causes of overdenture abutment loss. PMID- 17138347 TI - Smoking increased the risk of major complications of onlay bone graft. PMID- 17138348 TI - Skeletal calcium disturbances in heavy smokers may contribute to high salivary calcium concentration. PMID- 17138349 TI - Advances in clinical practice and continuing education "The precautionary context clinical practice model: a means to implement the evidence-based approach". PMID- 17138350 TI - Evidence-based decision making in dental practice. AB - For an evidence-based dental practice, shared decision making is made possible using an intuitive or analytic approach in reconciling best evidence and patient characteristics and preferences. Decision trees assist the dentist when presenting conditions and patient values are less certain in recommending treatment modalities whose benefits and harms are variable or unknown. Using clinical practice guidelines (or protocols organized within a decision tree), best evidence, based on the "average patient", is integrated into the decision tree. Probability, utility, and economic data are calculated and analyzed in providing the patient with informed and individualized choices of care. Evidence based decision making results in a process in which an individual patient is informed of the best available evidence and all relevant options. The outcome of the process is that patient utilities and cost concerns are fully addressed in optimizing the clinical decision. PMID- 17138351 TI - Oral lichenoid lesions associated with dental amalgam restorations: a 60-year-old woman with a 7-year history of painful, erosive mucosal lesions. PMID- 17138352 TI - Powered toothbrushes may reduce plaque and gingivitis at least as effectively as manual toothbrushing. PMID- 17138354 TI - Dental fear in children may be related to previous pain experience during dental treatment. PMID- 17138355 TI - Orthodontic repositioning of traumatically intruded maxillary incisors minimizes negative sequelae. PMID- 17138356 TI - The treatment of severe intrabony defects with GTR therapy may be used to maintain the CAL and retain teeth long term. PMID- 17138357 TI - A relatively low cumulative survival rate (CSR) over 5 years when endosseous tapered dental implants are placed either immediately after tooth removal or according to a delayed-placement protocol. PMID- 17138358 TI - Subantimicrobial dose doxycycline (SDD) improves probing parameters associated with periodontitis. PMID- 17138359 TI - Patient's ratings can be used to measure the satisfaction of different mandibular implant-retained overdentures. PMID- 17138360 TI - Clinical attachment level gain enhanced by addition of deproteinized bovine bone and barrier membrane to papilla preservation technique. PMID- 17138361 TI - Oral health-related quality of life is often poor among patients seeking third molar surgery. PMID- 17138362 TI - DNA sequence variations in the genes MSX1 and TGFA may lead to developmentally missing teeth. PMID- 17138363 TI - More than 5 full-mouth radiographic series increases intracranial meningioma risk. PMID- 17138364 TI - Patient and tooth specific risk indicators for posterior tooth fracture are identified. PMID- 17138365 TI - The prevalence of oral mucosal lesions in United States adults: data from the Third National Health and Nutrition Examination Survey, 1988-1994. PMID- 17138366 TI - Depression is associated with nonspecific physical symptoms in Asian TMD patients. PMID- 17138367 TI - Dental fluorosis increases caries risk. PMID- 17138369 TI - Model of evidence-based dental decision making. AB - Shared decision making empowers patients as informed consumers, helping them to make clinical decisions that optimize their personal oral health. Evidence-based dentistry supports this process by providing best evidence that, when presented in visual and interactive formats, focuses consultation time on treatment or therapy options and their trade-offs. Currently, an explosion of evidence and technological advancements have necessitated a cooperation that translates into an interdisciplinary approach to care delivery. For evidence-based dentistry, this interdisciplinary approach includes the interaction of researchers, clinicians, and patients in promoting individual patient care. To facilitate this interaction, a computerized model of evidence-based dental decision making is presented to manage knowledge in its application to clinical practice. This model includes the use of decision aids and a decision tree composed of multiple clinical practice guidelines. These aids allow for quantifying treatment options in terms of estimates using probability, utility, and cost data. With these estimates, decision analysis and the flexibility to manipulate data provide patients with increased control and acceptance of the decisions that they make about their personal oral health. PMID- 17138372 TI - Teacher-supervised toothbrushing with fluoride has little effect on dental caries prevention. PMID- 17138370 TI - Multiple factors associated with HA-coated implants had more marginal bone loss but greater survival rates at 12 years than pure titanium implants. PMID- 17138373 TI - Stainless steel crowns improve success rate of root canal treatment in primary teeth. PMID- 17138374 TI - Patients remain very satisfied with oral implants for a reasonably long time. PMID- 17138375 TI - Total fluoride intake higher in subjects of low socioeconomic status. PMID- 17138376 TI - Glass ionomer cement (GIC) sealants on first primary molars failed to reduce caries experience among Welsh preschoolers. PMID- 17138377 TI - Immediate loading of dental implants in selected cases may provide similar success rates as compared with delayed loading. PMID- 17138378 TI - Jawbone quality, jaw shape, implant length, and overdenture treatment protocol were associated with implant failures. PMID- 17138379 TI - More well-differentiated squamous cell carcinomas in individuals younger than 40 years old. PMID- 17138380 TI - Individual self-reported items are not accurate markers for a history of chronic periodontitis. PMID- 17138381 TI - Radiographic tips on predicting inferior alveolar nerve exposure. PMID- 17138382 TI - Visible third molars are associated with an increased risk for abnormal periodontal measures on adjacent second molars. PMID- 17138383 TI - Patients with epilepsy may have an increased risk of tooth loss. PMID- 17138384 TI - Premorbid traumatic stressors are highly prevalent in chronic TMD. PMID- 17138385 TI - Probing depth but not attachment level may be associated with the development of impaired glucose tolerance among (40- to 79-year-old) Hisayama residents. PMID- 17138389 TI - What does the precautionary principle mean for evidence-based dentistry? AB - The precautionary principle calls for preventive actions in the face of uncertain information about risks. It serves as a compass to better guide more health protective decisions in the face of complex risks. Applying precaution requires thinking more broadly about risks, taking an interdisciplinary approach to science and policy, and considering a wide range of alternatives to potentially harmful activities. While often criticized as antiscientific, the precautionary principle represents a challenge to scientists and public health professionals to develop newer and more effective tools for characterizing and preventing complex risks, in addition to being more explicit about uncertainties. This article examines the role and application of precaution in the context of dental practice, where activities that may convey risks also have public health benefits, and risk trade offs are a possibility. We conclude that the precautionary principle is not at odds with, but rather complements evidence based practice in situations of scientific uncertainty and complex risks. PMID- 17138390 TI - When can the result of epidemiologic research not eliminate the need to invoke the precautionary principle? PMID- 17138391 TI - FDA marketing claims, and the practitioner. AB - The Food and Drug Administration (FDA) is the federal agency that is tasked with regulating market entry for medical devices. The laws that govern this process are codified in the Federal Food Drug and Cosmetic Act (the Act) and the regulations are based on this law. The medical device amendments to the Act were instituted in 1976, instituting the methods for classification of medical devices and the format for the premarket review of devices. Information for practitioners on how medical devices come to market, what data are required, how specific claims are cleared, and how the practitioner can give input to the system are critical for the further development of safe and effective medical devices. PMID- 17138393 TI - The effect of patient-centered plaque control and periodontal maintenance therapy on adverse outcomes of periodontitis. AB - The purpose of this study was to evaluate systematic reviews that addressed the effectiveness of periodontal maintenance therapy for the management of patients with periodontitis. Recent surveys of dental care patterns suggest a marked increase in preventive and maintenance periodontal care in populations that retain the dentition for an increasingly longer lifetime. A considerable body of clinical investigation concludes that a multitherapy periodontal maintenance approach is effective in improving periodontal outcomes in patients treated for periodontitis. Individual components of such maintenance therapy were assessed, including the effects of an oral examination, personal oral hygiene instructions, supragingival scaling and polishing, subgingival scaling and root planing, adjunctive procedures, and maintenance frequency. There is much controversy about improvement in oral health that may accrue from the placebo effect of an examination and the maintenance ritual. Improved plaque control by the patient in anticipation of a forthcoming examination alone might be reflected in decreased measurements for plaque accumulation and gingival inflammation but the role of placebo effects on periodontitis remains unclear. There are insufficient randomized controlled trials to reach conclusions regarding the individual beneficial effects of repeated oral hygiene instructions or routine scaling/polishing on the recurrence of periodontitis. While subgingival root planing seems an effective component of periodontal maintenance, neither clinical investigations nor randomly controlled trial evidence have established an ideal maintenance frequency based on individual patient risk for periodontitis. The adjunctive beneficial effects of both locally and systemically administered antimicrobial agents were statistically significant for some formulations, and may be particularly useful clinically in patients who are resistant to mechanical therapy. We conclude that few clinical or randomized controlled studies have evaluated the individual benefit or required frequency of the periodontal maintenance ritual for patients who are relatively resistant or susceptible to periodontitis. PMID- 17138394 TI - Periodontitis and smoking: an evidence-based appraisal. PMID- 17138396 TI - The relationship between occlusion and TMD: an evidence-based discussion. PMID- 17138397 TI - Current evidence providing clarity in management of temporomandibular disorders: summary of a systematic review of randomized clinical trials for intra-oral appliances and occlusal therapies. PMID- 17138398 TI - TMJD 3: a genetic vulnerability disorder with strong CNS involvement. PMID- 17138400 TI - The educational system's integration of evidenced-based principles: what are the obstacles in the dental school environment? PMID- 17138401 TI - The importance of teaching critical thinking early in dental education: concept, flow and history of the NYU 4-year curriculum or "Miracle on 24th Street: the EBD version". PMID- 17138402 TI - Treatment plan for integrating evidence-based decision making into dental education. AB - The aims of this paper are to present the findings from the scientific literature that discuss strategies that can contribute to a "best practices" treatment plan model for effectively integrating Evidence-Based Decision Making (EBDM) into curricula. MEDLINE, CINAHL, and HealthSTAR databases were searched, as was the Cochrane Database of Systematic Reviews. Studies and articles, ranging from systematic reviews to articles proposing models and recommendations for how to implement EBDM into curricula and faculty development were reviewed. Several common themes emerged and form the basis for a treatment plan model. The first step in developing any treatment plan is a thorough assessment of the current situation or problem. Recognizing that there are multiple phases to the assessment of an educational system, the focus of this paper will be to understand which teaching and learning strategies are most effective. These, in turn, will inform faculty of needed curricular changes and skill development training, requisites in order for them to prepare students to be successful in providing patient care using the best available evidence. Elements of a suggested treatment plan will be presented with the caveat that each dental school will need to develop an implementation plan based on an assessment of its own environment and needs. PMID- 17138404 TI - Practicing with the evidence. PMID- 17138405 TI - The role of evidence in formulating public health programs to prevent oral disease and promote oral health in the United States. AB - A major goal of federal, state, and local health agencies is to reduce the burden of disease in populations. To obtain sufficient resources to achieve this goal, they must document the importance of the health problems addressed and the impact and efficiency of programs. CDC supports core activities within state and local health departments to promote health and prevent disease. This presentation will focus on the need for evidence to document the effectiveness of these activities that include routine monitoring of oral health, risk behaviors, and other factors; implementing effective population-based interventions; and evaluating programs to ensure successful translation of interventions. CDC supports research to build the evidence for innovative strategies to promote oral health in communities. This presentation should increase awareness of evidence-based tools and activities that are supported by CDC to strengthen public health practice within states. PMID- 17138407 TI - The evidence supporting alternative management strategies for early occlusal caries and suspected occlusal dentinal caries. AB - OBJECTIVE: To assess the strength of the evidence describing the effectiveness of alternative strategies to the detection and management of early occlusal caries and suspected occlusal dentinal caries. METHODS: Nine detection and intervention decision points were identified as being central to the management of early occlusal caries and suspected occlusal dentinal caries, or suspicious areas. For each decision point, the evidence for effectiveness was assessed, using existing systematic reviews when available, and nonsystematic review methods when necessary. RESULTS: For the 2 detection decisions (early occlusal caries and suspicious areas) the strength of the evidence was weak. Accuracy in detecting early occlusal caries was extremely variable within and across detection methods. Approximately 50% of suspicious areas identified had dentinal caries. The strength of the evidence for effectiveness of nonsurgical approaches for the management of early occlusal caries was weak for all 3 management strategies examined (doing nothing, sealants, remineralization). This evidence suggested that sealants were highly effective, with remineralization reflecting moderate effectiveness. For the management of suspicious areas, the strength of the evidence was still weaker and reflected the same relative effectiveness. For the surgical management strategy for suspicious areas, operative treatment, the evidence was strong and reflected high effectiveness for preventive resin restorations, but no evidence was available for minimally invasive techniques. CONCLUSION: Identification methods for early occlusal caries are not accurate. The strength of the evidence for effectiveness of nonsurgical management strategies for early occlusal caries is at best, weak. The available evidence suggests that sealing both enamel caries and suspected occlusal dentinal caries is the most effective management approach if subsequent maintenance of the sealed surfaces can be assured. PMID- 17138408 TI - Endodontics and implants, a catalog of therapeutic contrasts. AB - Dentists may be faced with the choice to retain a tooth by performing endodontic therapy and restoration or to extract the tooth and replace it with an implant and restoration. The purpose of this study was to catalog areas where implant and endodontic therapies differ so as to assist dentists in making treatment decisions and in identifying areas deserving of future research. Differences in diagnostic procedures and prognostic indicators were listed. With respect to treatment outcomes, study designs, success criteria, treatment results, systematic reviews, complications, clinician expertise, and the use of patient based measures were discussed. The need for clinically applicable consensus statements and treatment protocols was noted. It was concluded that at this time, choices between implant and endodontic therapies cannot be solely based on outcomes measurement evidence; that different modes of outcome measure frustrate direct comparison; that endodontic and implant therapies profoundly differ in many ways; that although rigorous and clearly defined outcome measures have been proposed for use in endodontic and implant outcomes studies, they are very rarely used; that long-term, large, clearly defined studies, with simple and clear outcome measures, for example survival in combination with defined treatment protocols, are needed to measure the clinical performance of endodontic and implant therapies; and it was recognized that broad outcomes data may not be sufficiently specific to directly impact clinical decision making. PMID- 17138410 TI - The ADA perspective. AB - BACKGROUND: The American Dental Association (ADA) has long relied on available scientific evidence in its commitment to use credible scientific data and analyses in policy-making, and its communications with the dental profession and the public. OVERVIEW: The evidence-based dentistry (EBD) process is a systematic approach that reviews and publishes the evidence relevant to specific clinical questions. The goal of this process is to help practitioners provide patients with quality oral health care. The ADA has developed a strategic plan of EBD initiatives and activities in order to increase the knowledge base and improve educational programs; to encourage significant collaboration on EBD-related issues from interested organizations, specialty groups, and various outside agencies; and to increasingly derive the best scientific evidence for use in clinical practice in concert with professional judgment and patient treatment preferences. CONCLUSION: The ADA endeavors to provide relevant information that can assist dentists in translating systematic-review findings and implementing an evidence-based approach in everyday clinical practice. This supports the Association's goal to continue supporting the best available evidence to assist health care teams in the provision of quality patient care and preventive techniques that improve oral health care outcomes. PMID- 17138411 TI - EB dentistry--a third-party perspective. PMID- 17138412 TI - Peer review and evidence-based dental practice. PMID- 17138413 TI - Real-world implementation of evidence-based dental practice. PMID- 17138414 TI - The challenges of transferring evidence-based dentistry into practice. AB - The goal of evidence-based dentistry is to help practitioners provide their patients with optimal care. This is achieved by integrating sound research evidence with personal clinical expertise and patient values to determine the best course of treatment. Although clinicians embrace this concept, its implementation in clinical practice has been slow. In this article, barriers against the implementation of evidence-based care are examined and possible solutions are offered. The dental profession is committed to providing the best possible dental care for patients. This is proving to be more complex due to a virtual "information explosion" on new therapies, techniques, and materials; increased consumer understanding of treatment possibilities and therapeutic outcomes; and changing sociodemographic patterns. Although the profession advocates the importance of evidence-based dental disease prevention and treatment, practitioners have been slow to implement this concept. In 2003, the California Dental Association (CDA) formulated an evidence-based dentistry (EBD) action plan that included the formation of a task force to monitor EBD efforts and implement programs to educate CDA members on this methodology. The challenges of transferring EBD into clinical practice were key issues addressed by the task force, and much of their deliberations and perspectives are reflected in this article. Possible solutions for eliminating barriers against evidence-based care will also be explored. PMID- 17138416 TI - The American Board of Dental Examiners clinical dental licensure examination: a strategy for evidence-based testing. AB - The evolution and implementation of a uniform national clinical examination has become a reality under the guidance of the American Board of Dental Examiners (ADEX). The defined purpose of this examination is to provide a defensible construct that assesses the critical clinical competencies of dental and dental hygiene candidates that will differentiate between competent and incompetent entry-level practitioners. The author includes a discussion of validity evidence that will be gathered by ADEX in order to support the decisions and interpretations made concerning competency and the agency's claim for validity. Included is a brief discussion of some controversial opinions regarding general validity concerns involving clinical dental licensure exam constructs. The author intends for this manuscript to provide a better appreciation for the science of psychometrics as it is applied to evidence-based testing for entry-level dental and dental hygiene candidates. PMID- 17138417 TI - The residency pathway to dental licensure: the paradigm shift from inception to policy. AB - Following decades of minimal advancement, the dental licensure reform movement has rallied around the recent introduction of a bold new concept. The New York State Dental Association has introduced and passed legislation that allows the substitution of a successfully completed dental residency program for the traditional clinical licensing examination. The residency must be one approved by the Commission on Dental Accreditation of the American Dental Association. New York's main objective was to elevate the standards required for licensure beyond those of minimal competency. The residency experience provides extensive didactic and clinical experience in a supervised setting relevant to contemporary dental practice. PMID- 17138418 TI - Interest and responsibility of ADA in dental licensure. AB - Initial licensure is a critical milestone and point of entry to the profession; it should go without saying that the organization that represents more than 70% of professionally active dentists would care deeply about the process that determines the character of its future and defines its image in the eyes of the public. The American Dental Association's (ADA) documented history of activity and leadership on licensure issues and the organization's guiding documents (Strategic Plan, Current Policies, Principles of Ethics and Code of Professional Conduct and Constitution and Bylaws) all lend credence to the Association's role in the licensure process. ADA members, other dental organizations, private and governmental agencies, and the public recognize the ADA as an authority on matters relating to dentistry. These circumstances comprise the best available evidence supporting the important role of the ADA in facilitating communication, collaboration and consensus-building in the continuous enhancement of the licensure process to meet the needs of all stakeholders. PMID- 17138419 TI - What the available evidence on clinical licensure exams shows. PMID- 17138420 TI - The evidence for immediate loading of implants. PMID- 17138421 TI - Practice-based research: is it worthwhile?...A clinician's view. PMID- 17138422 TI - Questional benefit from occlusal adjustment for TMD disorders. PMID- 17138424 TI - Fewer crowns rather than large amalgam restorations require additional treatment 10 years after placement. PMID- 17138425 TI - Ball-springs require more repair than bar-clip implant-supported mandibular dentures. PMID- 17138426 TI - Amalgams lead to more catastrophic failures in endodontically treated premolars with class II cavities. PMID- 17138427 TI - Phase 1 interceptive orthodontic treatment reduces the severity of malocclusion. PMID- 17138428 TI - Intraoral slow-release fluoride devices may decrease the incidence of dental caries in high-risk children. PMID- 17138429 TI - No benefit from regular 6-month fluoride gel applications in low-risk caries individuals. PMID- 17138430 TI - Genetics may have a significant contribution to dental caries while microbial acid production appears to be modulated by the environment. PMID- 17138431 TI - Losing teeth leads to an unhealthy diet associated with cardiovascular disease risk. PMID- 17138432 TI - Mood psychopathology is possibly associated with bruxism. PMID- 17138433 TI - Smoking and obesity may be independent risk factors for periodontitis. PMID- 17138434 TI - Down syndrome patients may exhibit poorer periodontal conditions. PMID- 17138435 TI - Low birth weight babies are not at increased risk of primary tooth caries. PMID- 17138436 TI - One in 3 removable denture users in the United States has denture stomatitis. PMID- 17138437 TI - Oral hygiene status and smoking are strong risk indicators of periodontal disease in Thai adults. PMID- 17138439 TI - Implementing evidence-based decision making in the private practice setting. Why do it? PMID- 17138440 TI - Implementing evidence-based decision-making: forming a student extracurricular evidence-based journal club. PMID- 17138441 TI - Only minor differences in implant performance exist between various implant types. PMID- 17138443 TI - Postoperative oral amoxicillin/clavulanic acid reduces inflammatory complications following extraction of impacted mandibular third molars. PMID- 17138444 TI - Sealed teeth get less restorative treatment. PMID- 17138445 TI - Locally delivered antimicrobials adjunctive to scaling and root planing provide additional PD reduction and CAL gain in the treatment of chronic periodontitis. PMID- 17138446 TI - Prosthetic status is related to quality of life. PMID- 17138447 TI - Dental health history forms may underreport adolescent smoking. PMID- 17138448 TI - In a resource-limited setting, oral visual screening may reduce oral cancer mortality. PMID- 17138449 TI - Exercise, high-quality diet, and maintaining normal weight are associated with reduced levels of periodontitis. PMID- 17138450 TI - Vertical alveolar bone loss is more prevalent among smokers. PMID- 17138451 TI - Morphological analysis of periapical radiographs in the mandible may identify women with low skeletal bone mineral density. PMID- 17138452 TI - Maternal periodontal disease is related to preterm low birth weight delivery in a group of Brazilian women. PMID- 17138453 TI - Genetics and environmental factors play important roles in the risk for periodontal disease and edentulism. PMID- 17138454 TI - Periodontal status is unrelated to preterm low birth weight in a group of Caucasian German women. PMID- 17138455 TI - Obesity is associated with deep periodontal pockets in Japanese women. PMID- 17138456 TI - Alcohol intake impacts severe periodontitis. PMID- 17138457 TI - Dental fear, dental attendance, and self-report of dental erosion appear to be altered in women with eating disorders. PMID- 17138460 TI - Re: Diverse components of the oral environment in attention-deficit hyperactivity disorder (ADHD) make it difficult to establish whether ADHD is a risk factor for dental caries. JEBD 2005;5:39-40-review of Broadbent et al (2004). PMID- 17138462 TI - The birth of Research in Social & Administrative Pharmacy: a sincere welcome to subscribers, readers, and authors. PMID- 17138463 TI - Judgment processes in older adults' drug benefit evaluations. AB - BACKGROUND: The Medicare Prescription Drug, Improvement and Modernization Act of 2003 will provide drug coverage to older adults through a variety of mechanisms, including stand-alone prescription drug benefits. Variation in cost-sharing and utilization controls is permitted, leading potentially to a wide variety of prescription benefit plans. However, little is known regarding the manner in which beneficiaries will integrate information to form plan evaluations. OBJECTIVES: The objectives of this study were to assess and compare the use of compensatory and configural judgment models in older adults' evaluations of drug benefit plans. METHODS: Three focus groups were conducted with a total of 19 seniors to elicit relevant drug plan attributes. A separate group of 32 seniors then judged the suitability of 48 drug benefit profiles based on these attributes. Within-subjects regression analysis was used to reveal each participant's judgment policy. RESULTS: Focus groups suggested that copayment, premium, deductible, formulary use, and mail-service use were relevant plan attributes. Regression analyses showed that while most participants used compensatory judgment models in evaluation formation, 12 (37.5%) subjects used conjunctive judgment models. CONCLUSIONS: Configural judgment processes are used frequently by older adults when evaluating drug benefit plans. Future research using more fine-grained techniques (eg, process tracing) might help further elucidate judgment processes in this context. PMID- 17138464 TI - An inquiry into medication meanings, illness, medication use, and the transformative potential of chronic illness among African Americans with hypertension. AB - BACKGROUND: Hypertension is a chronic illness with serious economic and clinical consequences. The asymptomatic nature of this condition contributes to the challenge of persistent medication use. OBJECTIVES: The objectives of this qualitative study were to explore perceptions of medication meanings, illness, identity, and biographical disruption among people with hypertension, and to examine how salient themes and categories relate to medication use. METHODS: In person interviews were conducted with 20 participants. Eligibility criteria included being 18 years or older, diagnosed with hypertension, and currently taking antihypertensive drug treatment. Interviews were tape-recorded and transcribed. Data were analyzed with grounded theory methodology using coding techniques and constant comparison. RESULTS: All participants were African American and most were between 45 and 64 years. Key themes including perceptions of the medication being effective, lifesaving, part of life, and a reminder of the regimen were found to have a positive impact on medication use. Themes including fear of side effects, fear of dependency, forgetting to take medication, the medication being a hassle, and the experience of medication related sexual side effects were found to be negatively related to medication use. Participants were cognizant of consequences of uncontrolled hypertension, and illness control was important to them. Biographical disruption was minimal. Taking medications and changing diet were the most significant changes in the lives of participants after being diagnosed with hypertension. Achievement of lifestyle modifications had a positive impact on identity. CONCLUSIONS: Findings contribute to our understanding of medication use behavior and have implications for patient education and counseling. PMID- 17138465 TI - The development and performance validation of a tool to assess patient anticoagulation knowledge. AB - BACKGROUND: The complexity of anticoagulation therapy requires a patient's command of anticoagulation-related knowledge to assist in maintaining optimal therapy and reducing adverse events. Verbal evaluations may overestimate the patient's comprehension of anticoagulation knowledge. OBJECTIVES: This first phase study developed and pilot tested the Anticoagulation Knowledge Assessment (AKA) instrument, and provided validity evidence to support the use of the AKA. The AKA is an assessment instrument that (1) pharmacists can use to obtain objective evidence of patient anticoagulation knowledge, (2) can provide pharmacists with direction for patient-specific medication management education, and (3) functions as a tool for continuous quality improvement in anticoagulation education. METHODS: Using objective measurement methods, a convenience sample of 60 English-speaking patients receiving services from an inner-city and suburban pharmacist managed anticoagulation clinics was used in conjunction with objective measurement methods. RESULTS: Rasch analysis of 32 multiple-choice items representing 10 anticoagulation educational content areas demonstrated misfit statistics of less than 1.2. All 60 patients demonstrated person misfit statistics of less than 1.3. The educational content area was well represented and distributed. CONCLUSIONS: Because the AKA performed well, the data support that information gained from the AKA will provide pharmacists with direction for anticoagulation management education that is targeted to each patient's specific needs. Additionally, responses demonstrated objective data about those components of practice that are being taught effectively. PMID- 17138466 TI - Treatment perceptions and attitudes of older human immunodeficiency virus infected adults. AB - OBJECTIVE: To apply the Health Belief Model (HBM) in assessing the association of health beliefs, perceived benefits, perceived barriers, alternative therapy use, and sexual risk behaviors in relation to the treatment of human immunodeficiency virus (HIV) in a group of older HIV-infected patients. METHODS: A convenience sample of 100 older (50 years and above) HIV-infected patients in 2 Washington, DC, clinics was enrolled. A cross-sectional methodology used structured interviews to investigate the association among antiretroviral adherence, use of alternative therapies, treatment perceptions, and risk behaviors. Student t tests were conducted to examine significant relationships between HBM perceptions and demographic characteristics. Logistic regressions were conducted to assess likelihood of antiretroviral and alternative therapy use. RESULTS: The majority of the participants were black and had a high school education. Although participants believed that HIV was a severe disease, they did not perceive themselves to be susceptible to early progression to acquired immunodeficiency syndrome. Participants believed that antiretrovirals were beneficial, and they were not particularly burdened by perceived costs of antiretrovirals. The perceived costs (or barriers) of antiretrovirals were inversely associated with CD4 count (r=-0.25, P=.01) and positively associated with viral load (r=0.33, P < .01). Overall 21% of patients in this population indicated use of alternative therapies. Patients who tended to have a higher perception of severity of HIV and a higher perceived burden in using antiretrovirals were more likely to use alternative therapy. There was no difference in HBM perceptions among antiretroviral users and nonusers. CONCLUSIONS: In general, the benefits of taking antiretrovirals were clear to most patients, and the same patients did not view access to antiretrovirals as a significant barrier to treatment. Many patients, although aware of the severity of HIV disease, were not seeking modifications to sexual behavior. Furthermore, the actual medication-taking behavior of these patients resulted in significant impacts to their clinical status. Study results can be applied in the development of specific interventions that are intended to decrease HIV transmission among older adults and to improve medication-taking behavior among those who are already infected with HIV. PMID- 17138467 TI - Patients' willingness to pay for pharmacist-provided menopause and hormone replacement therapy consultations. AB - OBJECTIVE: To determine what women would be willing to pay for pharmacist provided menopause and hormone replacement therapy consultations and the relationship between willingness to pay (WTP) and patients' perceptions of importance of resolving menopausal symptoms, their satisfaction with physician services, their perceptions of pharmacists' abilities, and income. METHODOLOGY: A convenience sample of 203 adult women using medications to treat menopause was recruited by their pharmacists. A self-administered survey was used to address the study objectives. The contingent valuation method was used to assess WTP in 4 scenarios (ie, 50% and 100% improvement in symptoms after a half-hour initial and a half-hour follow-up visit, respectively). Logistic regression analyses were used to assess the relationship between WTP and the independent variables. RESULTS: Two hundred and three surveys were returned resulting in a response rate of 41%. Across all 4 scenarios, at least 85% of women surveyed were willing to pay a minimum of 20 US dollars for pharmacist-provided menopause and hormone replacement therapy services. Respondents were willing to pay a median of 40 US dollars (for both 50% and 100% improvement) per half-hour for initial consultations, and 20 US dollars and 40 US dollars (50% and 100% improvement, respectively) per half-hour for follow-up consultations. Regression analyses revealed that across all 4 scenarios, patients' perceptions of pharmacists' abilities and income were significantly and positively (P < .05) related to WTP for menopause and hormone replacement therapy counseling. CONCLUSION: This study showed that most women surveyed were willing to pay for pharmacist-provided hormone replacement therapy and menopause counseling services and that the WTP amounts increased as patients' perceptions of pharmacists' abilities and income increased. If pharmacists can effectively educate the public about their capabilities and expertise, they may discover that they have a new niche opportunity. PMID- 17138468 TI - The Authority/Pharmacotherapy Care model: an explanatory model of the drug use process in primary care. AB - BACKGROUND: Drug utilization studies have proliferated and many variants exist. Few models have been presented that account for all of the different types of studies and approaches. PURPOSE: This article presents the Authority/Pharmacotherapy Care model, a structural-functional model of the drug use process that illustrates the factors involved in drug utilization and the relationships between factors. The concepts of authority and transfer of authority underlie the relationships. METHODS: The drug use process is presented at the microlevel from the viewpoint of an individual who requires treatment with prescription drugs. The various categories of activity/authority (ie, level of patient care) are those of the individual, physician, pharmacist, patient, and drug. Influencing factors, both internal and external, impact upon each level of care. Three aspects must be considered at each level, which are structures, processes, and outcomes, according to Donabedian's model. RESULTS: The result is a structural-functional model that depicts all of the major points in the drug use process, which might be used as a framework to categorize drug utilization studies. CONCLUSIONS: This model may be used to represent the drug use process, identify the types of drug use studies, determine pertinent factors involved in the process, understand the relationships between factors, and help in evaluating drug use. PMID- 17138469 TI - Accounting principles, revenue recognition, and the profitability of pharmacy benefit managers. AB - OBJECTIVES: To contrast pharmacy benefit management (PBM) companies' measured profitability by using two accounting standards. The first accounting standard is that which, under Generally Accepted Accounting Principles (GAAP), PBMs are currently allowed to employ. The second accounting standard, seemingly more congruent with the PBM business model, treats the PBM as an agent of the plan sponsor. DATA SOURCES: Financial Accounting Standards Board (FASB) Emerging Issues Task Force Issue 99-19, U.S. Securities and Exchange 10-K filings and financial accounting literature. SUMMARY: Under GAAP record keeping, the PBM industry profitability appears modest. Using currently applied GAAP, the PBM treats all payment from the plan sponsor as revenue and all payment to the pharmacy as revenue. However, the PBM functions, in practice, as an entity that passes-through money collected from one party (the sponsor) to other parties (dispensing pharmacies). Therefore, it would seem that the nature of PBM cash flows would be more accurately recorded as a pass-through entity. When the PBM is evaluated using an accounting method that recognizes the pass-through nature of its business, the PBM profit margin increases dramatically. CONCLUSION: Current GAAP standards make traditional financial statement analysis of PBMs unrevealing, and may hide genuinely outstanding financial performance. Investors, regulators, pharmacies, and the FASB all have an interest in moving to clarify this accounting anomaly. PMID- 17138470 TI - The role of hospital and community pharmacists in pharmacovigilance. AB - In this paper, we give an overview of the role pharmacists actually have and could have in collecting reports of adverse drug reactions (ADRs) and more widely in pharmacovigilance. In the literature, several ways are mentioned in which the pharmacist, both the community pharmacist and the hospital pharmacist, can contribute to the safe use of drugs. In addition to their responsibilities regarding drug dispensing and compliance, they can have a substantial role in ADR reporting. Especially, hospital pharmacists can play a significant role in ADR reporting because the most serious adverse drug events occur in hospitals, and ADRs account for a substantial proportion of hospital admissions. Community pharmacists, however, can also play an important role in ADR reporting. This is, for example, the case in the Netherlands where community pharmacists contribute substantially, both in numbers and in quality of ADR reports. The contribution of the pharmacist to pharmacovigilance should, however, not be limited to ADR reporting. The various pharmaceutical disciplines could also greatly enhance our understanding of the nature of ADRs. If those involved in pharmaceutical disciplines can rise to this challenge, they will significantly help deepen our insights into ADRs. PMID- 17138471 TI - The interaction of direct-to-consumer prescription drug advertising and stakeholders in the medication use process. PMID- 17138472 TI - The association of consumer cost-sharing and direct-to-consumer advertising with prescription drug use. AB - BACKGROUND: Previous research on the impact of various cost-sharing strategies on prescription drug use has not considered the impact of direct-to-consumer (DTC) advertising. OBJECTIVE: To explore the association of cost-containment strategies with prescription drug use and to determine if the association is moderated by DTC prescription drug advertising. METHODS: The study population included 288 280 employees and dependents aged 18 to 65 years with employer-sponsored health insurance contributing to the MEDSTAT MarketScan administrative data set. Person level enrollment and claims data were obtained for beneficiaries enrolled continuously during July 1997 through December 1998. Direct-to-consumer advertising data were obtained from Competitive Media Reporting and linked to the MEDSTAT enrollment files. Localized DTC advertising expenditures for one class of medication were evaluated and matched with prescription claims for eligible MEDSTAT contributors. The association of various types and levels of cost-sharing incentives with incident product use was evaluated, controlling for the level of DTC advertising, health status, and other demographic covariates. RESULTS: The relationship of cost-sharing amounts with drug use was modified by the level of DTC advertising in a geographic market. This relationship was dependent on the type of cost-sharing, distinguishing between co-payments for provider visits and co-payments for prescription drugs. Compared with low-advertising markets, individuals residing in markets with high levels of advertising and paying provider co-payments of $10.00 or more were more likely to use the advertised product. In the same markets, higher prescription drug co-payments were associated with a decreased likelihood of using the advertised product. A similar relationship was not observed for the nonadvertised competitor. CONCLUSIONS: Among insured individuals, response to cost-sharing strategies is moderated by DTC prescription drug advertising. The relative ability of cost-sharing strategies to influence drug use should be interpreted with caution in the presence of DTC advertising. PMID- 17138473 TI - TV commercials for prescription drugs: a discourse analytic perspective. AB - BACKGROUND: The US Food and Drug Administration has called for research that may assist in developing standards for risk/benefit messages in the promotion of prescription drugs. Linguistics-based models of meaning and inference, though frequently applied to advertising, have not hitherto been used in this arena. OBJECTIVE: This study was intended to illustrate how discourse analysis, a methodology for microanalysis of texts in context, can elucidate the workings and interplay of promotional, informational, and other functions of direct-to consumer drug advertising, anticipating threats to "fair balance" and pinpointing textual phenomena and issues suited to empirical study. METHODS: The text and visuals of a small corpus were analyzed along several dimensions, using theoretical insights of linguistic pragmatics and ethnography of speech to ask what the advertisement is seeking to do and what messages a viewer is likely to derive. RESULTS: The linguistic and rhetorical features include an intense switching and fusion of styles and modalities: the traditional advertising distinction between personal and impersonal, "company" and "consumer", was ostentatiously flouted. The role of spokesperson was assigned to characters in a real or virtual narrative. The narrative portion of the text and images often struck an ironic or postmodern note, eg, by mixing science with science fiction. The overall functions of the commercials (promotional, informational, and aesthetic) were themselves frequently blended. The text deployed several linguistic or rhetorical strategies to send a double message for promotional advantage, including syntactic-semantic ambiguity, voice-over risk messages at odds with upbeat visuals, and a vagueness of certain words in particular contexts. CONCLUSIONS: Findings contribute to our understanding of how TV commercials convey meaning with respect to drug benefits and risks, with implications for advertisers, regulators, and patient education. They also suggest new foci for empirical study. PMID- 17138474 TI - Television advertisement format and the provision of risk information about prescription drug products. AB - BACKGROUND: Considerable attention has been afforded to analyzing the content of and assessing consumers' reaction to print direct-to-consumer drug ads, but not so for televised ads. OBJECTIVE: To determine whether advertisements with different risk severity and risk presentation would significantly affect viewers' (1) recall of information contained in the advertisement, (2) evaluation of the advertisement, and (3) perceptions of the advertised product's risks. METHODS: Data were collected from a sample of 135 first-year pharmacy students at a Midwestern college of pharmacy. After viewing 1 of the 6 advertisements designed for this study, participants were asked to complete a self-administered survey. Chi-square and analysis of variance were used to analyze the data. A 2x3 between subjects design was used to test the effects of 2 levels of risk severity (high- vs low-risk severity) and 3 levels of risk presentation (original ad containing integrated risk message, deintegrated risk message/dual modality using male voice over, deintegrated risk message/dual modality using female voice-over). RESULTS: Results of analysis of variance procedures revealed that deintegrating risk information by placing it at the end of the advertisement and the use of captions in addition to oral messages (dual modality) (1) improved the recall of general and specific side effect information, (2) led to a perception that the advertisement had greater informational content, (3) resulted in lower Advertisement Distraction, and (4) lessened cognitive and affective aspects of information overload for the advertisement containing the high-risk severity medication. However, this pattern of findings was not found for the low-risk severity medication. CONCLUSION: Alternative methods for presenting risk information in direct-to-consumer ads affected some aspects of information recall and advertisement evaluation, but were not shown to affect risk perceptions regarding the advertised products. PMID- 17138475 TI - Direct-to-consumer advertising and the patient-physician relationship. AB - BACKGROUND: Differences in attitudes of patients and physicians toward direct-to consumer advertising (DTCA) of prescription medications may influence the patient physician relationship, which may in turn influence health care outcomes. OBJECTIVE: The objective of this study was to provide empirical evidence to show how the patient-physician relationship may be influenced by DTCA. METHODS: The study used a cross-sectional research design. Pharmacists at 71 pharmacies were asked to distribute self-report survey instruments to patients who used either a proton-pump inhibitor or a nonsedating antihistamine by prescription. Data were collected between March and June 2003. Hierarchical multiple regression was used to assess the research propositions. RESULTS: A usable response rate of 7.3% (n=326) was obtained. About 95% of the sample had seen an advertisement for either a proton-pump inhibitor or a nonsedating antihistamine. Response to DTCA was significantly related to the patient's satisfaction with the physician and the patient's evaluation of communication, even after controlling for demographic variables and length of the patient-physician relationship. Other outcome variables (trust, relationship commitment, and average visit time) were not significantly related to DTCA response. Post hoc analyses revealed that DTCA responders, who asked their doctor to prescribe a specific product after seeing its advertisement, rated satisfaction and communication lower than the DTCA nonresponders. Additional follow-up analyses showed that these findings were largely attributed to patients in the proton-pump inhibitor group. CONCLUSION: The findings suggest that response to DTCA may be significantly related to patients' evaluation of communication quality and their satisfaction with the physician. However, these findings may not generalize to all medical conditions or types of patient. Furthermore, DTCA response does not appear to be related to other patient-physician relationship outcome variables. Given these findings, key variables to explore in future DTCA research with respect to its effect on the patient-physician relationship include patient expectations and disease/condition type. PMID- 17138476 TI - Distinguishing characteristics of patients who seek more information or request a prescription in response to direct-to-consumer advertisements. AB - OBJECTIVE: The objective of this study was to examine demographic and psychographic profiles of individuals who sought additional information or requested a prescription drug based on a direct-to-consumer advertisement. METHODS: A cross-sectional descriptive survey was used for collecting data from a random sample of 200 Minnesotans during Fall 2002. Chi-square and Mann-Whitney U tests were used as nonparametric tests for assessing differences in distributions between our categories of study subjects. RESULTS: Out of 177 deliverable surveys, 81 (46%) were returned. Of these, 80 surveys were usable for analysis. The results showed that the distinguishing characteristics of individuals who sought additional information based on an advertisement were associated with demographic variables such as number of drugs taken daily and monthly out-of pocket expenditures for prescription drugs. In contrast, distinguishing characteristics of individuals who requested prescription drugs (in addition to seeking information) based on an advertisement were psychographic in nature such as (1) viewing themselves as having greater influence on their physician, (2) having a stronger relationship with their physician, (3) expressing greater satisfaction with their current therapy, (4) viewing prescriptions as less of a burden, and (5) having higher outcome expectations for prescription drugs compared to the respondents who did not ask for a prescription drug based on a direct-to-consumer advertisement. CONCLUSIONS: Distinguishing characteristics of information seekers were demographic in nature, whereas those characteristics of prescription requesters were psychographic in nature. PMID- 17138477 TI - Drug information-seeking intention and behavior after exposure to direct-to consumer advertisement of prescription drugs. AB - BACKGROUND: Concerns about direct-to-consumer advertisement's (DTCA's) information quality have raised interest in patients' drug information-seeking after DTCA exposure. OBJECTIVE: To identify predictors of patients' intentions and behaviors to seek drug information from physicians, pharmacists, and the Internet after DTCA exposure, using theories of planned behavior and self efficacy. METHODS: One thousand patients were randomly selected from 3,000 nationwide osteoarthritic patients. A self-administered survey examined predictors of intention including measurements of attitude toward behavior, subjective norm, perceived difficulty, self-efficacy, controllability, self identity, intention, exposure to ads, and control variables. After 6 weeks, another survey measured respondents' information-seeking behavior. For patients exposed to DTCA, 6 multiple regressions were performed for information-seeking intention and behavior for 3 information sources: physicians, pharmacists, and the Internet. RESULTS: The response rates were 61.9% and 80.1% for the first survey and the second survey, respectively. Four hundred and fifty-four participants reported exposure to DTCA about arthritis prescription medicines in the previous month. Over 41% of the variance in intention and over 18% of the variance in behavior were explained by the regression procedures. The consistent positive predictors of intention were attitude toward behavior, self-identity, attitude toward DTCAs of arthritis medication, and osteoarthritis pain; while the consistent positive predictors of behavior were intention and osteoarthritis pain. The strongest predictors of intention were self-identity for physicians, subjective norm for pharmacists, and attitude toward behavior for the Internet. Perceived difficulty and self-efficacy did not predict intention, and self efficacy and controllability did not predict behavior. CONCLUSIONS: DTCA-prompted drug information-seeking may be under patients' complete volitional control. To promote information searching, efforts could be made to affect factors predicting intention. Interventions could address patients' attitude toward behavior, the influence of their important others, and their role as information seeker, respectively, for information sources like the Internet, pharmacists, and physicians. PMID- 17138478 TI - Using social cognitive theory to explain consumers' behavioral intentions in response to direct-to-consumer prescription drug advertising. AB - BACKGROUND: Previous research describing consumers' communication behaviors in response to direct-to-consumer advertising (DTCA) suggests a social cognitive rationale to explain DTCA-related communication behavior. OBJECTIVE: Guided by social cognitive theory, the objective of this study was to explore outcome expectancy and self-efficacy beliefs as predictors of individuals' intentions to communicate with their physicians about an advertised drug. METHODS: One hundred and seven female college students completed a questionnaire, read an advertisement for an oral contraceptive drug, and completed a second questionnaire. The questionnaires assessed participants' self-efficacy and outcome expectancy beliefs, intended communication behavior, and demographic information. RESULTS: Pearson product-moment correlation analyses showed that outcome expectancy (r=0.75, P<.01) and self-efficacy (r=0.21, P<.05) beliefs were associated positively with intentions to communicate with physicians in response to DTCA. However, ordinary least squares regression analyses revealed that only outcome expectancy beliefs predicted intended communication behavior (B=1.56, P<.01). Results also showed that participants had a relatively greater likelihood of requesting information about, than requesting a prescription for, the advertised drug [t(106)=14.75, P<.01]. CONCLUSIONS: The results identify cognitive factors that guide consumers' plans for interacting with physicians in response to DTCA. Health care providers can use these results to guide communication with patients regarding DTCA and meet patients' drug-related informational expectations. PMID- 17138479 TI - Direct-to-consumer advertising via the Internet: the role of Web site design. AB - BACKGROUND: Recent attempts to propose criteria for judging the quality of pharmaceutical and healthcare Web sites do not distinguish between attributes of Web site design related to content and other attributes not related to the content. OBJECTIVES: The Elaboration Likelihood Model from persuasion literature is used as a framework for investigating the effects of Web site design on consequents like attitude and knowledge acquisition. METHODS: A between-subjects, 2 (high or low involvement)x2 (Web site designed with high or low aspects of visual appeal) factorial design was used in this research. RESULTS: College students were randomly assigned to these treatment groups yielding a balanced design with 29 observations per treatment cell. Analysis of variance results for the effects of involvement and Web site design on attitude and knowledge indicated that the interaction between the independent variables was not significant in both analyses. Examination of main effects revealed that participants who viewed the Web site with higher visual appeal actually had slightly lower knowledge scores (6.32) than those who viewed the Web site with lower visual appeal (7.03, F(1,112)=3.827, P=.053). CONCLUSIONS: Results of this research seem to indicate that aspects of Web site design (namely aspects of visual appeal and quality) may not play a role in attaining desired promotional objectives, which can include development of favorable attitudes toward the product and facilitating knowledge acquisition. PMID- 17138480 TI - Introduction of direct-to-consumer advertising of prescription drugs in Canada: an opinion survey on regulatory policy. AB - BACKGROUND: Canada is strongly influenced by US cross-border direct-to-consumer advertising (DTCA) and has held consultations to discuss introduction of DTCA since 1996. This article describes a survey of Canadian drug policy experts carried out in 2001, during one such legislative review. The survey results are compared to more recent DTCA policy developments. METHODS: We recruited key informants on pharmaceutical policy to complete a faxed questionnaire that queried their opinions on DTCA information quality, effects on drug and health care use, and regulatory issues. Respondents were asked about the evidence they had used to back their opinions. Analysis was descriptive. RESULTS AND DISCUSSION: Of 79 identified potential participants, 60 (76%) participated, 40% of whom were from federal and provincial government; 3% were private insurers; 18%, 15%, and 8% were from health professional groups, consumer groups, and patient groups, respectively; 8% and 7% were from pharmaceutical and advertising industries, respectively. Opinions were highly polarized on the effects of DTCA on drug and health care use. Advertising and pharmaceutical industry respondents were generally positive, public sector, health professional and consumer groups generally negative. Over 80% believed DTCA leads to higher private and public drug costs and more frequent physician visits. Fewer judged billboards or television to be appropriate media for DTCA than magazines or the Internet, and most believed that children and adolescents should not be targeted. CONCLUSION: Given the polarization observed within this survey, we examined how DTCA policy has evolved in Canada since 2001. The federal government has legislative authority over DTCA, but bears few of the additional costs potentially incurred through policy change. These fall to the provinces, which provide an eroding patchwork of public coverage for prescription drugs in the face of rapidly increasing costs. No new federal legislation has been tabled since 2001. However, considerable shifts in administrative policy have occurred, all supportive of expanded advertising. Thus, the law continues to be restrictive but its application less so. PMID- 17138481 TI - Methodological challenges surrounding direct-to-consumer advertising research- the measurement conundrum. AB - OBJECTIVES: Numerous studies have focused on the impact of direct-to-consumer (DTC) prescription drug advertising on consumer behavior and health outcomes. These studies have used various approaches to assess exposure to prescription drug advertising and to measure the subsequent effects of such advertisements. The objectives of this article are to (1) discuss measurement challenges involved in DTC advertising research, (2) summarize measurement approaches commonly identified in the literature, and (3) discuss contamination, time to action, and endogeneity as specific problems in measurement design and application. METHODS: We conducted a review of the professional literature to identify illustrative approaches to advertising measurement. Specifically, our review of the literature focused on measurement of DTC advertising exposure and effect. We used the hierarchy-of-effects model to guide our discussion of processing and communication effects. Other effects were characterized as target audience action, sales, market share, and profit. RESULTS: Overall, existing studies have used a variety of approaches to measure advertising exposure and effect, yet the ability of measures to produce a valid and reliable understanding of the effects of DTC advertising can be improved. Our review provides a framework for conceptualizing DTC measurement, and can be used to identify gaps in the literature not sufficiently addressed by existing measures. CONCLUSIONS: Researchers should continue to explore correlations between exposure and effect of DTC advertising, but are obliged to improve and validate measurement in this area. PMID- 17138482 TI - The study of direct-to-consumer advertising for prescription drugs. AB - OBJECTIVES: The objectives of this article are to (1) identify key methodological issues related to investigating the effects of direct-to-consumer advertising (DTCA) for prescription drugs, (2) highlight opportunities and challenges that these issues pose, and (3) provide suggestions to address these challenges and opportunities from a social and administrative pharmacy perspective. METHODS: Through a review of existing literature and consultation with research colleagues, we identified 3 broad issues regarding the study of DTCA for prescription drugs: (1) the importance of problem formulation, (2) the role of health behavior and decision-making perspectives, and (3) data collection and data analysis challenges and opportunities. Based upon our findings, we developed recommendations for future research in this area. RESULTS: Clear problem formulation will be instructive for prioritizing research needs and for determining the role that health behavior and decision-making perspectives can serve in DTCA research. In addition, it appears that cluster bias, nonlinear relationships, mediating/moderating effects, time effects, acquiescent response, and case mix are particularly salient challenges for the DTCA research domain. CONCLUSIONS: We suggest that problem formulation, selection of sound theories upon which to base research, and data collection and data analysis challenges are key methodological issues related to investigating the effects of DTCA for prescription drugs. PMID- 17138483 TI - Relationships and responsibilities are critical to team care in medication management. PMID- 17138484 TI - Adherence to therapeutic regimens. PMID- 17138485 TI - The relationship of guideline-concordant depression treatment and patient adherence to oral diabetes medications. AB - BACKGROUND: Many patients with diabetes experience depression, yet it is unclear if the treatment of depression in diabetic patients is concordant with national guidelines, and whether appropriate antidepressant use is associated with better diabetes self-care behaviors. OBJECTIVES: The purpose of this study was to (1) determine whether antidepressant medication use for managed care enrollees with type 2 diabetes was concordant with The Agency for Healthcare Research and Quality depression treatment guidelines; and (2) examine the relationship between guideline concordance and oral diabetes medication adherence. METHODS: Retrospective analyses were conducted using medical/pharmacy claims for 2001 from a managed care organization in the midwestern United States. Subjects were adults with type 2 diabetes treated with oral medications only. The subjects were divided into 3 groups: (1) guideline-concordant users of antidepressants; (2) those who received antidepressants not in concordance with the guidelines; and (3) nonusers of antidepressants. Antidepressant users were determined to be in concordance with the acute phase treatment guidelines if they filled at least 90 days supply of antidepressant drugs within 118 days of the first fill. Adherence to diabetes medications was measured by the medication possession ratio. A 1-way analysis of variance with Scheffe's test was used to compare the antihyperglycemic medication possession ratio across the three groups. RESULTS: One hundred eighty-two (12.5%) of the 1454 subjects initiated treatment with antidepressants. Eighty-nine (48.9%) of the 182 antidepressant users were in concordance with the acute phase treatment guidelines. Subjects with subconcordant antidepressant use had a lower mean diabetes medication possession ratio than those with either guideline-concordant use or no use (F=14.3, P<.01). CONCLUSION: Over half of the diabetic patients initiating treatment for depression did not receive therapy in concordance with the Agency for Healthcare Research and Quality guidelines. Patients whose antidepressant use was not concordant with the guidelines were also less adherent to diabetes medications. PMID- 17138486 TI - Patient decision making: strategies for diabetes diet adherence intervention. AB - BACKGROUND: Patient self-care is critical in controlling diabetes and its complications. Lack of diet adherence is a particular challenge to effective diabetes intervention. The Transtheoretical Model (TTM) of Change, decision making theory, and self-efficacy have contributed to successful tailoring of interventions in many target behaviors. OBJECTIVE: The purpose of this study was to develop a diagnostic tool, including TTM measures for the stages of change, decisional balance, and self-efficacy, that pharmacists involved in diabetes intervention can use for patients resistant to a diet regimen. METHODS: A questionnaire was developed through a literature review, interviews with diabetic patients, an expert panel input, and pretesting. Cross-sectional implementation of the questionnaire among a convenience sample of 193 type 1 and type 2 diabetic patients took place at 4 patient care sites throughout the southeastern United States. Validated measures were used to collect respondent self-report for the TTM variables and for demographic and diabetes history variables. Social desirability was also assessed. RESULTS: Relationships among TTM measures for diet adherence generally replicated those established for other target behaviors. Salient items were identified as potential facilitators (decisional balance pros) or barriers (decisional balance cons and self-efficacy tempting situations) to change. Social desirability exhibited a statistically significant relationship with patient report of diet adherence, with statistically significant differences in mean social desirability across race categories. CONCLUSIONS: The TTM measures for the stages of change, decisional balance, and self-efficacy are useful for making decisions on individually tailored interventions for diet adherence, with caution asserted about the potential of diabetes patients to self-report the target behavior in a socially desirable manner. Future research directions, implications, and limitations of the findings are also presented. PMID- 17138487 TI - Collaborative medication management in a team-based primary care practice: an explanatory conceptual framework. AB - BACKGROUND: In ambulatory practice many different health care professionals are involved in the drug use process. The roles and functions of these individuals can be difficult to define, because of a lack of a common ground for discussion. Deliberating the topic is important for developing a team approach to medication management. OBJECTIVE: To describe a conceptual framework to create a platform that can be used by different health care providers to identify, define, and discuss roles and responsibilities in collaborative medication management. METHODS: Authors reviewed and reflected upon their experiences as practitioners in implementing a pharmaceutical care-based consulting practice within a family medicine practice setting. Key roles and responsibilities relative to collaborative management of medications were identified and described. RESULTS: A conceptual framework subsequently developed, the Team Approach to Medication Management, consists of 3 primary components referred to as medication-related practices (medication prescribing, medication taking, and medication dispensing). Each of these primary practices is supported by a team of health care professionals who have supportive roles and responsibilities. In the Team Approach to Medication Management framework, the patient's medication-taking practice holds a central and key position within a collaborative approach to medication management. CONCLUSIONS: The proposed Team Approach to Medication Management framework can be used to guide discussions and decisions among the different health care providers working in primary care to define both direct and indirect roles that health care practitioners and patients play in collaborative medication management. PMID- 17138488 TI - Pharmacy-specific quality indicators for asthma therapy. AB - BACKGROUND: Lack of appropriate drug therapy in asthma patients is a recognized quality problem leading to preventable emergency room visits, hospitalizations, or death. While indicators measuring pharmacotherapy quality on the level of prescribers and third party payers are widely used, no such indicators exist for pharmacies. OBJECTIVE: This study aimed to (1) develop quality indicators for asthma care applicable to retail pharmacies, (2) estimate the prevalence of inappropriate asthma drug therapy, and (3) explore variation in the quality of care across pharmacies. METHODS: We present a descriptive analysis of automated patient-specific dispensing data from February 1, 2000 to January 31, 2001 of beta-agonists and anti-inflammatory agents with a Food and Drug Administration approved indication for asthma. Two quality indicators of potentially inappropriate drug therapy were applied: (1) the proportion of patients who obtained more than a 360-day supply (assuming maximum acceptable daily dose) of short-term beta-agonists (SABA) including all inhaler types, inhaler solutions, and syrups during the 12-month study and (2) the proportion of patients with 2 consecutive early refills of defined SABAs. Indicator values with 95% confidence intervals are reported for each pharmacy. RESULTS: Pharmacies had an average of 328 (range 169-534) patients who received SABAs. An average of 11 patients per pharmacy (3.4% of all patients who received SABAs) met the indicator 1 definition. The second indicator identified 8.2% (27) patients per pharmacy as short-term SABA overusers (range 3.9-11.9%). Of these, 48% did not receive any anti-inflammatory agents during the time frame when SABA overuse occurred. CONCLUSION: Application of drug therapy quality indicators at the level of individual pharmacies using dispensing data is feasible and identifies opportunities for quality improvement. Indicator 2 is most appropriate for daily practice, because it allows for timely identification of potentially uncontrolled patients, and offers a balance between indicator sensitivity and positive predictive value. PMID- 17138489 TI - Polypharmacy trends in office visits by the elderly in the United States, 1990 and 2000. AB - BACKGROUND: Polypharmacy has been extensively studied internationally and reported to be increasing among the elderly. Within the United States, few studies have examined polypharmacy trends in the elderly population and even fewer studies addressed those at-risk for polypharmacy. OBJECTIVES: To examine the trends in office-based visits in the United States by the elderly involving polypharmacy and identify elderly at-risk for polypharmacy. METHODS: Data from the 1990 and 2000 National Ambulatory Medical Care Surveys were used to examine polypharmacy visit trends in the elderly. The Bonferroni inequality method was used to analyze the visit estimates and visit rates. Logistic regression analysis was used to model predisposing, enabling, and need factors associated with polypharmacy visits in the elderly using the 2000 survey data. RESULTS: Office visits involving polypharmacy for elderly patients were estimated to have nearly quadrupled from 10.1 million in 1990 to 37.5 million in 2000. The proportion of visits by elderly patients involving polypharmacy was 7% in 1990 and 19% in 2000. The increase was consistent among all demographic groups and remained significant even after controlling for elderly population increase. Medication classes involved in polypharmacy remained consistent during the study period and included cardiovascular, hormonal, pain, and gastrointestinal medications. Analysis of the 2000 survey data revealed that several need (multiple diagnoses, chronic problems, and specific disease states), predisposing (female gender), and enabling factors (primary care provider visit and health insurance coverage) were associated with polypharmacy visits in the elderly. CONCLUSIONS: The study found a significant increase in elderly patients' office visits involving polypharmacy in the United States. The study also found that several need, predisposing, and enabling factors were associated with polypharmacy visits in the elderly. These findings suggest opportunities to review and manage elderly patients' medications as recommended by Healthy People 2010, a national agenda to improve the health of Americans. PMID- 17138490 TI - Comparison of potentially hepatotoxic drugs among major US drug compendia. AB - BACKGROUND: Although a large number of drugs include warnings or listed adverse reactions that describe reports of associated hepatotoxicity, the hepatotoxic risk is documented with different definitions in major drug compendia. OBJECTIVES: The purposes of this study were to compare inclusion of potentially hepatotoxic drugs, and analyze the ratings of hepatotoxic risk among major drug compendia. METHODS: To assess the risk of drug-associated hepatotoxicity, we used current literature of epidemiological studies and developed a 4-level rating scale of hepatotoxic drugs: 3, clear literature evidence of life-threatening hepatotoxicity; 2, multiple case reports or significant liver injuries; 1, no significant liver damage has been reported; and 0, no information. All drugs were evaluated using the 5 major US drug compendia: American Hospital Formulary Service (AHFS), United States Pharmacopeia Drug Information (USPDI), Facts and Comparisons (F&C), Physicians' Desk Reference (PDR), and Clinical Pharmacology (CP). Average rating scores were calculated as the sum of each drug rating score divided by the total number of drugs. One-way analysis of variance and independent t tests were conducted to compare the difference among the rating scores. RESULTS: In total, 175 different drugs and 3 therapeutic classes with hepatotoxic effects were identified in the compendia, including 59 antineoplastics, 28 anti-infectives, 17 nonsteroidal anti-inflammatory drugs, 17 antipsychotics or phenothiazine derivatives, 9 angiotensin-converting enzyme inhibitors, 6 anticonvulsants, 4 histamine-2 receptor antagonists, and other drugs. Average rating scores were 1.65 for AHFS, 1.10 for USPDI, 1.27 for F&C, 1.34 for PDR, and 1.61 for CP (F=7.93, P<.0001). The risk categories were significantly different among compendia in 4 therapeutic classes of antipsychotics and/or phenothiazines (F=3.471, P=.011), nonsteroidal anti inflammatory drugs (F=7.866, P<.0001), antineoplastics (F=2.476, P=.044), anti infectives (F=2.003, P=.098), and angiotensin-converting enzyme inhibitors (F=38.125, P<.0001). CONCLUSIONS: Rating scores of hepatotoxicity were significantly different among drug compendia. The different compendium put different emphasis on hepatotoxicity severity. Comprehensive evaluations of hepatotoxic-related drugs provide critical information for health practitioners. PMID- 17138492 TI - Role theory: literature review and implications for patient-pharmacist interactions. AB - BACKGROUND: There is a need to describe how role theory has been used by researchers to describe various phenomena in pharmacy so as to identify gaps in knowledge and future research priorities. OBJECTIVE: The primary purpose of this article is to review how role theory has been used in the community pharmacy literature. Secondary objectives are to (1) examine the use of role theory over the evolution of the profession of pharmacy, (2) determine what role theory perspectives have been used, (3) explore the implications of role theory for patient-pharmacist interactions, and (4) explore implications for future pharmacy research using role theory. METHODS: A literature search of Web of Knowledge and International Pharmaceutical Abstracts was conducted. Articles were included if they examined some aspect of role theory in community pharmacy and were categorized based on their role theory perspective and "era" of pharmacy in which they were published. RESULTS: Thirty research articles were identified spanning from 1956 to 2004 with the majority (19 studies) using mail surveys. Articles used functionalist (9 studies), organizational (7), functional and organization (4), cognitive (10), and symbolic interactionist (1) perspectives to role theory. The number of articles using role theory has been increasing over time. The functional and symbolic interactionist perspectives provide rich descriptions of the multiple pharmacy roles and allow for a clearer understanding of the barriers affecting actors' experience. Using an organization perspective, role stressors such as role conflict, ambiguity, and overload were found to impact pharmacists' worklife. Cognitive role theory research has clearly shown that pharmacists' and patients' expectations for the encounter shape interaction. CONCLUSIONS: The use of role theory informs both researchers and practitioners about the practice of pharmacy and patient interactions. Still, there is much work to be done in role theory and pharmacy research. Experimental designs, longitudinal studies, and qualitative research methodologies may warrant greater use and attention. Role change should be identified as a priority, and research is needed to elucidate what interventions change patients' and pharmacists' expectations of the patient pharmacist interaction. PMID- 17138493 TI - The mediating role of health beliefs in the relationship between depressive symptoms and medication adherence in persons with diabetes. AB - BACKGROUND: Although bivariate relationships between depressive symptoms, health beliefs, and medication adherence have been identified, the complex relationship among these 3 constructs has not been explicated. OBJECTIVE: This study examines the mediating role of patients' beliefs about diabetes and diabetes medications in relation to depressive symptoms and diabetes medication adherence. METHODS: A survey was sent to 1700 persons with type 2 diabetes who were enrolled in a managed care organization in the United States. The bivariate relationships between depressive symptoms, diabetes-related health beliefs, and diabetes medication adherence were assessed. A structural equation model was developed to determine if health beliefs mediated the relationship between depressive symptoms and medication adherence. RESULTS: Usable responses were received from 445 subjects. Greater depressive symptoms were associated with lower adherence to diabetes medications. The structural equation model indicated that the effect of depressive symptoms on medication adherence was mediated through perceived side effect barriers, perceived general barriers, and self-efficacy. Patients with severe depressive symptoms perceived more barriers to treatment adherence and were less confident in their ability to adhere to medication. In turn, reduced self-efficacy and heightened perceived barriers had a negative association with patients' adherence to diabetes medication regimens. CONCLUSIONS: Decreased adherence to diabetes medications in patients with both diabetes and depressive symptoms may be partly explained by the association of depression with patients' beliefs about diabetes medications and their self-efficacy for medication use. PMID- 17138494 TI - Perspectives on Drug and Therapeutics Committee policy implementation. AB - BACKGROUND: Drug and therapeutics committees (DTCs) are expected to make difficult decisions that may have an impact on the clinical and economic outcomes of drug use. There have been few studies investigating the barriers to DTC policy implementation, and little is known about ways to improve the process. OBJECTIVES: The specific objectives of this qualitative study were to explore stakeholder opinions with respect to (1) the perceptions of barriers to Drug and Therapeutics Committee (DTC) policy implementation and (2) ways to improve DTC policy implementation. METHODS: Stakeholders of Australian DTCs participated in focus group discussions. Discussions were audiotaped and transcribed verbatim. Thematic content analysis was conducted. RESULTS: Six focus group discussions were conducted. A number of barriers were identified (lack of resources, lack of follow-up, lack of ownership, low DTC profile within the organization, and overreliance on pharmacy to implement policy). Participants were of the opinion that prioritizing decisions, optimizing pharmacy roles, provision of real-time information, and active procurement of organizational commitment were some of the ways to improve DTC policy implementation. CONCLUSIONS: Although this study was conducted in an Australian setting, the challenges facing DTCs in other countries are likely to be similar. Ownership, social influence, and resources are important issues in the implementation of policy/guidelines for many DTCs, both in Australia and internationally. It is expected that these issues will have an impact on policy implementation. Therefore, the findings of this study may be widely applicable. This work reinforced the notion that the significance of DTCs lies beyond decision making alone. The time and expertise invested in decision making could be undermined, if DTC policies are not effectively implemented. PMID- 17138495 TI - Understanding practice change in community pharmacy: a qualitative study in Australia. AB - BACKGROUND: Much of the research on cognitive pharmaceutical services has focused on understanding or changing community pharmacist behaviour, with few studies focusing on the pharmacy as the unit of analysis or considering the whole profession as an organisation. OBJECTIVES: To investigate practice change and identify facilitators of this process in community pharmacy, with specific focus on the implementation of cognitive pharmaceutical services (CPS) and related programs. METHODS: Thirty-six in-depth, semistructured interviews were conducted with participants from 2 groups, community pharmacies and pharmacy "strategists," in Australia. The interview guide was based on a framework of organizational theory, with 5 subject areas: roles and goals of participants in relation to practice change; experiences with CPS; change strategies used; networks important to the change process; and business impacts of CPS. Interviews were transcribed verbatim and thematically content analyzed, using NVivo software for data management. RESULTS: Five key themes relating to the change process were derived from the interviews: change strategies (process- and behaviorally oriented); social networks (within and beyond the pharmacy); drivers of change (eg, government policy); motivators (eg, professional satisfaction); and facilitators of practice change (remuneration for implementation or service delivery, communication and teamwork, leadership, task delegation, external support or assistance, and reorganization of structure and function). CONCLUSION: The use of an organizational perspective yielded rich data from which an understanding of the practice change process in relation to CPS implementation was gained. Current programs for the implementation and delivery of CPS have not taken into account all of the factors that have the ability to facilitate change in community pharmacy. Not only do future programs need to be underpinned by these elements, but policy makers must include them when planning remuneration and dissemination strategies. PMID- 17138496 TI - Factors affecting collaborative care between pharmacists and physicians. AB - BACKGROUND: To have a positive impact on patient outcomes achieved with drug therapy, it is likely that pharmacists will work more closely with physicians to manage medications collaboratively. Yet, little is known about the factors that will support such collaborative care between pharmacists and physicians. OBJECTIVE: The objective of this study was to identify significant influences on collaborative care between pharmacists and physicians, from the perspective of pharmacists. METHODS: Data were collected through a survey mailed to a national sample of 321 pharmacists identified by state pharmacy associations as being innovative practitioners. Three types of influences were assessed: individual characteristics, contextual factors, and exchange characteristics. Individual characteristics included demographics and a personality measure. Context variables included practice environment and professional interactions between pharmacists and physicians. Exchange characteristics were trustworthiness, role specification, and relationship initiation. Four items asked about the pharmacist's collaborative care with a physician. A hierarchical linear regression analysis was performed with collaborative care as the dependent variable and the individual, context, and exchange characteristics as the independent variables. RESULTS: One hundred sixty-six usable surveys (53.4%) were returned. About 64% of the respondents were male, with a mean age of 43.7 (SD+/ 11.2) years. Linear regression analysis of the complete model produced an R(2)=0.805 (P<.001). Significant predictors in the model included the context variable, professional interaction, and the exchange characteristics, trustworthiness and role specification. CONCLUSION: Overall, the collaborative working relationship model largely explained collaborative care between pharmacists and physicians. Researchers are encouraged to use these findings when studying pharmacist-physician collaboration. In addition, pharmacists seeking to work with physicians should attend to developing trustworthiness and clarifying their clinical roles with physicians. PMID- 17138497 TI - Use of health-related quality of life information in managed care formulary decision-making. AB - BACKGROUND: The extent to which the increased volume of available health-related quality of life (HRQOL) information and heightened education has increased the acceptance and use of HRQOL remains unclear. Likewise, the value of HRQOL information in the formulary decision-making process continues to be undefined. OBJECTIVE: To investigate the perceptions and use of HRQOL by managed care decision-makers in the formulary development process. METHODS: A mail survey was sent to a nationwide sample of 108 Academy of Managed Care Pharmacy (AMCP) members who were involved in formulary management. Survey candidates were identified according to their job titles listed in the 1999-2000 AMCP membership directory. The survey process began in May 2000 and ended in August 2000. The main outcome measures included (a) managed care formulary decision-makers' assessment of HRQOL as a treatment outcome, (b) the existing role and future use of HRQOL information in formulary decisions, and (c) the level of understanding of HRQOL concepts and the benefits attributable to favorable HRQOL results. RESULTS: A response rate of 51.9% was obtained. Most of the respondents (>70%) believed that patients consider HRQOL as an important treatment outcome. Fewer respondents (43%) felt that payers view HRQOL outcomes as an important quality indicator. Most respondents (95%) considered HRQOL data in making formulary decisions, and many (73%) believe that HRQOL outcomes will play a more important role in future formulary decisions. Respondents indicated a better understanding of disease-specific and generic HRQOL measurements than utility measurement and interpretation of results. A minority of respondents (34%) would be willing to pay a higher price for a product with better HRQOL outcomes. When asked which factors would lead to increased use of HRQOL information, respondents indicated that health care cost savings and increased productivity were considered important (77% and 65%, respectively). CONCLUSIONS: A drug product with better HRQOL outcomes alone will not command a favorable listing on managed care formularies. HRQOL information needs to be made more applicable to managed care decision-making. Future studies should focus on the link between positive HRQOL outcomes, health care cost savings, and increased productivity. PMID- 17138498 TI - Testing a pharmacist-patient relationship quality model among older persons with diabetes. AB - BACKGROUND: Considering recent changes to the Medicare program, pharmacists will have unique opportunities to be reimbursed for providing Medication Therapy Management Services to older persons with diabetes. A high-quality pharmacist patient relationship can lay the foundation for effective provision of Medication Therapy Management Services and improved care in this cohort. OBJECTIVE: To test a pharmacist-patient relationship quality model in a group of older persons with diabetes from the patient's perspective. Antecedents to relationship quality were pharmacist participative behavior/patient-centeredness of relationship, patient participative behavior, and pharmacist-patient interpersonal communication. Pharmacist-patient relationship commitment was the outcome of relationship quality studied. METHODS: Data were collected via mailed questionnaire from a random sample of 600 community-dwelling adults in the United States who (1) were 65 years of age and older, (2) had type 1 or type 2 diabetes, (3) used at least one prescription medication to treat their diabetes, and (4) used some type of nonmail order pharmacy as their primary source of obtaining prescription medications. Model relationships were tested using path analysis. RESULTS: The adjusted response rate was 41.6% (221/531). The models explained 47% and 49% of the variance in relationship quality and relationship commitment, respectively. In the relationship quality model, pharmacist participative behavior/patient centeredness of relationship (beta=.51, P<.001) and pharmacist-patient interpersonal communication (beta=.17, P=.008) had direct effects on relationship quality. In the relationship commitment model, relationship quality had a direct effect on relationship commitment (beta=.60, P<.001). Pharmacist participative behavior/patient-centeredness and pharmacist-patient interpersonal communication had indirect effects on relationship commitment through their effects on relationship quality, which is a mediator in the model. CONCLUSIONS: Results affirm findings from previous research showing that patients' perceptions of pharmacist participative behavior/patient-centeredness of relationship and pharmacist-patient interpersonal communication are positively related to perceptions of relationship quality. Also, relationship quality is a strong mediator between pharmacist participative behavior/patient-centeredness of relationship and relationship commitment, as well as between pharmacist-patient interpersonal communication and relationship commitment. PMID- 17138499 TI - Consumption and costs of antihypertensive drugs in Mexico: are diuretic agents a standing technological trajectory? AB - BACKGROUND: Little is known about hypertension medication consumption and costs in Mexico. Hypertension control is a pharmacological challenge and a public health issue. OBJECTIVE: (a) To compare drug sales, number of written prescriptions, and monthly treatment costs among 5 classes of antihypertensive drugs and (b) to analyze diuretic drug sales and prescriptions to determine whether these antihypertensive agents represent an established technological trajectory. METHODS: A retrospective time series data study from 1999 to 2003. Data sources used were International Marketing Services of Mexico drug sales and the Mexico Prescription Audit databases. The 5 different classes of antihypertensive drugs were accommodated into 4 main technological trajectories according to their main biological mechanisms of action. Each technological trajectory was assessed using consumption and prescription data. Daily defined dose was used to calculate drug treatment costs. RESULTS: The market for cardiovascular agents is one of the largest, and in 2003 accounted for a value market share of 59 billion US dollar and a unit share of 40.7 million. Among cardiovascular agents, antihypertensive drugs made up a large percentage of market shares. Calcium channel blockers and angiotensin-converting enzyme inhibitors I had the biggest share value of the total cardiovascular market. Amlodipine had the highest share among calcium channel blockers, and enalapril and captopril had the largest share among angiotensin-converting enzyme inhibitors I. The top-selling diuretic drug was furosemide. The trend in number of prescriptions was parallel to that in sales. The diuretic spironolactone was the most expensive drug treatment (59 US dollar). Treatment with spironolactone might represent 47% of the income of a Mexican family if their household income was close to minimum wage (124 US dollar). CONCLUSIONS: The most effective and least expensive drugs-diuretics-had the smallest market share of all antihypertensive agents in Mexico. Nevertheless, diuretic agents are still in use and kept over time a steady market share both in value and in units. PMID- 17138500 TI - Using the Health Belief Model to test factors affecting patient retention in diabetes-related pharmaceutical care services. AB - BACKGROUND: Diabetes is one of the deadliest and most costly diseases. Attrition rates among patients in diabetes management programs may preclude optimal success. Theoretical models, such as the Health Belief Model, may be useful for identifying factors responsible for patients' continued enrollment in such programs. OBJECTIVES: (1) To design and test a reliable and valid survey instrument for assessing patients' perceptions of diabetes-related pharmaceutical care services. (2) To determine factors affecting patient retention in pharmaceutical care services. METHODS: This cross-sectional exploratory study used convenience sampling to survey type 1 and 2 diabetes patients receiving pharmaceutical care at 25 Eckerd Pharmacy sites in Florida. Survey items were designed using constructs from the Health Belief Model. Reliability (Cronbach's alpha) and validity (exploratory factor analysis) were assessed. Independent t tests, Pearson's correlation coefficients, and multiple regression analysis were conducted. RESULTS: Seventy usable surveys were returned (47%). In the absence of pharmaceutical care, patients felt susceptible to at least one of 8 diabetes related conditions (hypoglycemia, hyperglycemia, diabetic foot infections, eye problems, cardiovascular risks, blood pressure problems, cholesterol problems, and kidney diseases). For these conditions, about 48% to 95% of patients perceived that their threat had reduced because of pharmaceutical care. Accordingly, more than half perceived the services as beneficial, with counseling for blood sugar monitoring rated as the most beneficial. All respondents rated the services as helpful, and 64 intended to continue regular utilization. Overall helpfulness of the service and patient retention were positively correlated (r=0.33, P<.00). Perceived susceptibility predicted perceived threat reduction (R(2)=0.22, P<.01). Perceived threat reduction, blood sugar monitoring, and overall helpfulness of the service predicted patient retention in the service (R(2)=0.41, P<.00). Factor analysis extracted 4 factors: perceived threat reduction, perceived susceptibility, perceived benefits, and blood sugar monitoring. Cronbach's alphas for the scales ranged from 0.63 to 0.91. CONCLUSION: Two key constructs of the Health Belief Model influence use of diabetes-related pharmaceutical care services: perceived susceptibility and threat reduction. In an effort to increase patient retention, pharmacists need to assess patient perceptions and structure their services to address patient perceptions and concerns. PMID- 17138501 TI - Task analysis of patients' medication-taking practice and the role of making sense: a grounded theory study. AB - BACKGROUND: Patients on long-term medications have varied medication-taking practices and complex and often unmet medication information needs. OBJECTIVES: The objective of this qualitative study was to describe from the patient's perspective the medication-taking tasks performed by patients currently receiving long-term medications and then to hypothesize how these tasks relate to patients' medication information needs. METHODS: In-depth semistructured interviews were conducted with individuals between the ages of 18 and 65, who had a college or university education, and who were on at least one long-term medication. Grounded theory approach was used for data gathering and data analysis. Maximum variation and theoretical sampling were used and the sample size was determined when theoretical saturation was reached in the core category. Interpretive and theoretical validity were ensured through member checking, through the use of the constant comparative method, and by a review of the results by a panel of pharmacists and physicians. RESULTS: Ten participants aged between 41 and 64 years were included in the study sample. The participants had between one and 7 chronic illnesses, duration of these illnesses to date varied from 1 year to 40 years, and each participant was taking between one and 13 medications. A model was developed that consists of 4 thematic categories: (a) making sense of medication taking, (b) medication-taking acts, (c) mediation-taking self assessment, and (d) context of medication taking. The main category was making sense of medication taking that consisted of 3 subcategories: (a) nonproblematic mode, (b) problematic mode, and (c) stunned mode. The model explains how and why a patients' need for medication-taking education may vary because their medication-taking practices changes. The model also connects each category to medication information that people may need. CONCLUSIONS: Findings contribute to our understanding of medication-taking practice of individuals on long-term medication and have implication for patient education. PMID- 17138502 TI - Willingness to pay for praziquantel treatment in a hyperendemic community of Ogun State, Nigeria. AB - BACKGROUND: Schistosomiasis is highly endemic in Nigeria. However, very little is known about the usage and social marketing of praziquantel in the control of schistosomiasis in endemic communities and on the persons willingness to pay for the drug to support its extensive use and thus aid control efforts in Nigeria. OBJECTIVE: The study examined the willingness of people in a hyperendemic community, to pay for treatment with praziquantel before and after provision of the drug, with a view to assess the feasibility of achieving widespread coverage. METHODS: Data were collected through focus group discussions among household heads and other adult family members and through the use of a questionnaire to survey household heads in Imala-Odo, a community hyperendemic for schistosomiasis, in southwest Nigeria. The main outcome measure was the respondents' willingness to pay for schistosomiasis treatment. RESULTS: The results showed that in contrast to the 92.3% of respondents who expressed willingness to buy the drug in the preintervention study, only 46.5% actually purchased the drug during intervention for the treatment of their infected household members. The respondents' level of education, occupation, and income influenced their willingness to pay (P<.05). Statistical tests showed that those willing to pay for treatment had a higher income score than those unwilling to do so. Most respondents preferred their community head and reliable persons chosen by the community as convenient outlets for drug distribution. The number of household members respondents were willing to treat increased from 4 to 6 at N320.00 (US $2.52) in the preintervention and postintervention phases, respectively. Gender factor was found to influence the respondents' perceived average treatment cost; the females wanted N300.00 (US $2.36) against N100.00 (US $0.79) among the males. CONCLUSION: Efforts need to be made and sustained to ensure that all families can afford to praziquantel drug to achieve the ultimate goal of controlling the infection in endemic communities of Nigeria. PMID- 17138503 TI - A pilot study to describe antidepressant prescriptions dispensed to veterans after stroke. AB - BACKGROUND: Stroke is the third leading cause of death in the United States, following only heart disease and cancer. Approximately 500,000 strokes occur each year in the United States. Patients suffering from poststroke depression have higher rates of morbidity and mortality, slower physical recovery, and lower functional status than stroke patients without depression. Depressed stroke patients may benefit from early treatment with an antidepressant. OBJECTIVES: The objectives of this study were to (1) describe antidepressant-dispensing patterns to veteran patients during the 6-month period before their stroke and 1-year time afterward and (2) examine the association of poststroke antidepressant treatment with mortality. METHODS: Veteran patients (n=543) with one or more hospital discharges during FY2001 with a primary diagnosis of stroke were identified. The discharge date minus the length of stay indicated the index hospitalization date. Antidepressant prescription dispensing records were obtained from the Veterans Integrated System Network 8 Veteran's Affairs (VA) Pharmacy Benefits Management database. RESULTS: One-third of eligible veteran patients were dispensed one or more antidepressant prescriptions (n=181) either before or after the index hospitalization. Of these veterans, 146 received an antidepressant after the index hospitalization. More than 80% of veteran patients survived the stroke for 1 year or longer (n=448). Among the veteran patients who were discharged from the index hospitalization alive, those with no antidepressant or an antidepressant dispensed only before the stroke had a higher 1 year all cause mortality rate (8.0%) compared with those who were dispensed a prescription for one or more antidepressants afterward (4.8%). CONCLUSIONS: Veterans dispensed an antidepressant after their stroke appeared to have benefited. Opportunities to reduce poststroke sequelae and improve recovery may be lost. PMID- 17138504 TI - Pharmacy Service Orientation: a measure of organizational culture in pharmacy practice sites. AB - BACKGROUND: The importance of organizational culture in shaping everyday organizational life is well accepted, but little work has focused on organizational culture in pharmacy. Examining new pharmacists' experiences at various practice sites may help us to understand how these shape their professional ethos and practice habits. OBJECTIVES: (1) Present development and assessment of the Pharmacy Service Orientation (PSO) measure, a tool for assessing pharmacists' impressions of pharmacy practice sites. (2) Use data gathered from a sample of new pharmacists to explore potential predictors of PSO, including type of practice site, type of pharmacy work experience, and type of pharmacy degree. METHODS: Mail survey of randomly selected class of 1999 pharmacy graduates within 3 months of graduation (response rate: 259 of 1,850; 14%), each of whom reported on up to 6 different pharmacy practice sites for a total of 1,192 pharmacy observations. Pharmacy Service Orientation is scored on a 1-10 semantic differential scale and reliability was assessed using Cronbach's alpha. Predictors of PSO were explored using t test and ordinary least squares regression procedures. RESULTS: Reliability of the PSO across all observations was 0.86. When divided according to recency of experience and type of experience, reliabilities ranged from 0.78 to 0.87. Analysis of potential predictors of PSO showed that non-corporate-community sites had significantly greater pharmaceutical care-oriented cultures (mean PSOs of 7.42 and 5.13, respectively; P<.001). The same pattern was seen for academic and nonacademic worksites (mean PSOs of 7.46 and 6.01, respectively; P<.001). The pharmacist's pharmacy degree type was not predictive of PSO. Multivariate regression results showed that type of practice site and type of pharmacy work experience explained more than 25% of the observed variance in PSO. CONCLUSIONS: Pharmacy Service Orientation is a reliable measure. Statistically significant differences in PSO comparisons by degree and by experience type are explained by significant differences between the PSOs of corporate-community and non-corporate-community sites. PMID- 17138505 TI - Effects of collaborative drug therapy management on patients' perceptions of care and health-related quality of life. AB - BACKGROUND: It has been demonstrated that collaborative drug therapy management may result in enhanced medication adherence and improved clinical outcomes. It is not yet known whether CDTM is associated with patients' perceptions of care or self-reports of health-related quality of life. OBJECTIVES: Examine the impact of collaborative drug therapy management (CDTM) on patients' perceptions of care and health-related quality of life in 15 ambulatory clinics (6 intervention, 9 comparison) in the Fairview system of Minneapolis-St Paul, Minn. METHODS: The intervention was medication therapy management provided by pharmacists in collaboration with physicians (CDTM) for a 12-month period. Subjects were selected by age, gender, and presence of one of 12 medical conditions in the intervention (n=285) and comparison (n=285) group of patients. Comparison patients received usual care while intervention patients received at least 2 CDTM encounters. The CAHPS (formerly called the Consumer Assessment of Health Plans) 2.0 survey was administered to both the intervention and comparison groups poststudy to analyze patients' perceptions of care. The Short Form-12 (SF-12v2) was administered to intervention group patients pre-CDTM and 6 months post-CDTM to measure health-related quality of life in the intervention group. RESULTS: Differences in CAHPS scores were not statistically significant (P>.05), although there was a trend toward higher ratings of patients' personal doctor/nurse and doctors' communication in the CDTM intervention group relative to the comparison group. Physical role, social functioning, and physical component summary scales of the SF-12v2 improved significantly (P=.001, P=.014, and P=.024, respectively; P< or =.025 level). CONCLUSIONS: A trend toward improvements in patient perceptions of effectiveness of care using CAHPS suggests a need for further study. Health-related quality of life improvements in this study meet or exceed previous results incorporating pharmacists into primary care. Intensity and integration of CDTM services may be an explanation; however, prepost study design limits inferences. PMID- 17138506 TI - Addressing the issue of channeling bias in observational studies with propensity scores analysis. AB - Randomized Clinical Trials (RCTs) remain the gold standard for determining the utility of pharmaceuticals especially from a safety and efficacy standpoint. However, restrictive entry criteria and stringent protocols can be barriers to generalizing RCT findings to real world practices and outcomes. Observational studies overcome these limitations of RCTs since they are representative of real world populations and practices. Nonetheless, attributing causality remains a major limitation in observational studies, due to the non-random assignment of subjects to treatment. Non-random assignment can lead to imbalances in risk factors between the groups being compared and thus bias the estimates of the treatment effect. Non-random assignment can be particularly problematic in observational studies comparing older versus newer pharmaceuticals from similar therapeutic classes due to the phenomenon of channeling. Channeling occurs when drug therapies with similar indications are preferentially prescribed to groups of patients with varying baseline prognoses. In this manuscript we discuss the phenomenon of channeling and the use of a statistical technique known an propensity scores analysis which potentially adjusts for the effects of channeling. During the course of this manuscript we discuss tests for determining the quality of the derived propensity score, various techniques for utilizing propensity scores, and also the potential limitations of this technique. With the increasing availability of high quality pharmaceutical and medical claims data for use in observational studies, increased attention must be given to analytic techniques that adjust optimally for non-random assignment and resulting channeling bias. For research studies using observational study designs, propensity score analysis offers a reasonable solution to address the limitation of non-random assignment, especially when RCTs are too costly, time-consuming or not ethically feasible. PMID- 17138507 TI - Conceptualizing and measuring pharmacist-patient communication: a review of published studies. AB - BACKGROUND: Pharmacist-patient communication in community pharmacies has been studied for over 25 years with little effort to evaluate this research comprehensively. OBJECTIVE: The main objective of this review is to examine and summarize how researchers have conceptualized, defined, and measured pharmacist patient communication across studies and identify gaps in the literature. METHODS: Articles were compiled from a search of (1) Medline, IPA, CINAHL, and PubMed databases using the keywords, "counseling", "patient communication", "patient counseling", "patient education", "patient consult( *)", and/or "pharmacists", (2) bibliographies of selected articles. The search generated 56 studies on community pharmacy, of which 39 studies met the inclusion criteria. RESULTS: Most studies (72%) have used the term patient counseling, although pharmacist-patient communication and patient education were also used. The definition of patient counseling varies across studies. Almost half of the studies (49%) conceptualized pharmacist-patient communication solely as a pharmacist information provision activity. A total of 16 studies (41%) also focused on pharmacists' interpersonal behavior in addition to the information provision activity of the pharmacist. In contrast, patient communication behavior and the exchange process between both parties has been understudied. A total of 16 studies (41%) used a retrospective design. All studies used a cross-sectional design, with varying modes of data collection such as mail surveys, telephone interviews, nonparticipant observation, and shopper studies. Taped encounters are rare. SUMMARY/IMPLICATIONS: This review revealed that most studies have focused on a one way communication of pharmacists to patients. A need for examining the patient-pharmacist dyad is apparent. Future research could explore a greater use of taped encounters to analyze the interactive communication process, affective components of communication such as collaborative problem solving, interpersonal relationship development, and the expertise that patients bring into the encounter. PMID- 17138508 TI - Patients' evaluation of the appropriateness of their hypertension management--a qualitative study. AB - BACKGROUND: The existing appropriateness measures for prescribing used in the United States and the United Kingdom use clinical attributes. Treatment and care from a patient's perspective need to be evaluated in terms of whether they are more likely to lead to an outcome of a life worth living, in social, psychological, and physical terms. However, it is unclear whether patients specifically evaluate their prescribed medication and treatment. If so, do they use only clinical attributes or a combination of clinical and nonclinical attributes? OBJECTIVES: The aim of this study was to explore if patients evaluated their hypertension management, and if they did, investigate what attributes were involved in the evaluation. METHODS: Semistructured interviews, which focused on personal experiences of hypertension and its management were undertaken with patients (n=28). The aim of the interviews was to obtain, in a narrative format, the experiences, beliefs, and information that patients considered important when discussing the management of hypertension. Data analysis used a constant comparative method. RESULTS: All patients considered their hypertension management regimen appropriate, but were able to mention only 2 categories of attributes to justify their decision (the relationship with their General Practitioner and lowering of their blood pressure). Further series attributes were mentioned by the patient during the course of their interview; these attributes were considered to be involved in their evaluation. These implicit attributes were categorized as anxieties and concerns regarding treatment and diagnosis, explanation of the consequences of treatment, choice of antihypertensives, and the side effects experienced. CONCLUSIONS: Patient's evaluation of appropriateness was constructed from both explicit and implicit attributes. Implicit attributes, those not consciously known to the patient still, could be involved in the process of evaluating hypertension, its treatment, and care. Although the nonmedical attributes that are considered by patients can be categorized, it has to be remembered that it is the inherent meaning held by each individual patient involved when an evaluation is made. PMID- 17138509 TI - Exploring subjective outcomes perceived by patients receiving a pharmaceutical care service. AB - BACKGROUND: There is a need for evidence to support expanded pharmacist services, especially regarding subjective outcomes experienced by patients. However, it is unclear what aspects of life are actually affected by such services. OBJECTIVES: The aims of this study are to identify patients' perceptions of both an expanded pharmacist service and its impact on subjective outcomes such as health and quality of life, and to elicit variables that may be targeted in future evaluation of the impact of such services. METHODS: In-depth interviews were performed with 12 respondents receiving a patient medication record service. The interviews were analyzed using a qualitative constant comparative method. RESULTS: The service was perceived very differently by unique respondents in regards to its purpose and content, as well as its outcomes. It was difficult for respondents to describe the service as separate from other health care experiences. Gaining control of drug treatment and a perceived feeling of increased safety were central concepts in patients' description of the service. Talking spontaneously about health effects of the service carried no meaning for the respondents. CONCLUSIONS: The value of enhanced patient consultation, empowerment, the feeling of safety, and increased drug knowledge are important to patients receiving this kind of service and should be central in choosing outcome measures when studying such services. PMID- 17138510 TI - Drug insurance instability and its correlates: results from the 2000 Medical Expenditure Panel Survey. AB - BACKGROUND: Health insurance instability (ie, temporal gaps in health insurance coverage) is a prevalent phenomenon in the United States. To date, most studies have focused on the factors that affect the intermittent lack of health insurance coverage. However, no studies known to the authors have examined the factors associated with prescription drug insurance instability (ie, temporal gaps in drug insurance coverage) among working-age adults. Developing an accurate profile of persons with unstable drug insurance is essential to formulate rational policy to address this problem. OBJECTIVES: The objectives of this study were to (1) document the prevalence of prescription insurance instability among working-age adults and (2) describe the association between prescription drug insurance instability and demographic, socioeconomic status, and employment characteristics. METHODS: The data source used in this study was the 2000 Medical Expenditure Panel Survey. This study used a cross-sectional design using data provided by respondents at each of the 3 interviews conducted during the year 2000. Chi-square and hierarchical multinomial logistic regression analyses were used to describe the associations among (1) demographics, (2) socioeconomic status, and (3) employment characteristics and drug insurance status (classified as continuous, absent, or unstable). RESULTS: During the year 2000, 12.5% (21.1 million) of the working-age adults in the United States had unstable prescription drug coverage. Persons aged 35-54 years had lower rates of drug insurance instability compared with those aged 18-24 [adjusted odds ratio 0.66 (95% confidence interval 0.54-0.80)]. The least educated (12 or fewer years of education) were more likely than those with more education (13-16 years) to experience at least one period without drug coverage (62% vs 32%, P<0.01). The poorest respondents (those at less than 200% of the federal poverty level) were more likely than the wealthiest respondents (those at more than 400% of the poverty level) to report at least some time without drug coverage (37% vs 28%, P<0.01). Those experiencing a divorce or death of a spouse were more than twice as likely as stably married persons to experience at least one period without drug insurance [adjusted odds ratio 2.23 (95% confidence interval 1.68-2.96)]. Adults who were unstably employed during the year and/or who worked for small firms generally experienced higher rates of drug insurance instability. CONCLUSIONS: Prescription drug insurance instability is a prevalent phenomenon among working-age adults in the United States, with approximately 1 in 8 experiencing this problem during 2000. Our results suggest that demographics, socioeconomic status, and employment characteristics all play important roles in predicting prescription drug insurance status, with the least educated and poorest being particularly vulnerable to interruptions in drug coverage. Premium assistance programs providing subsidies to small firms' low-income employees and permitting small firms to form insurance pools may help to decrease the number of drug coverage uninsurance spells in this population. PMID- 17138511 TI - Influences on consumer spending for herbal products. AB - BACKGROUND: Despite the rapid growth in consumer spending on herbal products, we know little about factors that influence such spending. OBJECTIVE: To use a model of adoption to investigate consumers' spending on herbal products. METHODS: The study used a mail survey of a stratified random sample of 1,300 consumers. The population consisted of consumers aged 18 years and older residing in the United States. The sampling frame was a mailing list purchased from KM Lists. The independent variables were consumer characteristics, social systems, communication channels, and herbal characteristics. The amount of spending on herbs was the dependent measure, with responses divided into monthly spending of 10 dollars or less and more than 10 dollars . Binary logistic regression was performed to investigate the association between adoption model variables and spending on herbs. RESULTS: Of the 1,300 mailed surveys, there were 77 undeliverable surveys and 456 usable returned surveys, yielding a usable response rate of 37.3%. A total of 181 (39.7%) respondents reported using herbal products. The logistic regression was performed using the 168 herbal users who reported that they spent money in the past month on herbal products. The overall regression model was significant (P<.05, Nagelkerke R(2)=0.499). The significant influences on spending on herbals were age, over-the-counter (OTC) drug use, and use of an herb professional as an information source about herbs. Older people reported spending more on herbal products than younger people. OTC drug use was positively related to spending on herbals and appears to complement herbal usage. Finally, consumers who obtain information about herbals from an herb professional tend to spend more on herbals. CONCLUSION: An adoption model may be useful in explaining consumers' spending on herbal products. PMID- 17138512 TI - Relationships between Hispanic ethnicity and attitudes and beliefs toward herbal medicine use among older adults. AB - BACKGROUND: Little is known about the underlying intentions to use herbal medicines among Hispanic older adults. Understanding these intentions is critical to the provision of effective counseling. OBJECTIVES: The objectives were to (1) identify predictors of the intention to use herbal medicines for health problems in the next 6 months among Hispanic and non-Hispanic older adults using the Theory of Planned Behavior (TPB); and (2) compare their beliefs underlying significant predictors of intention with use herbal medicines for health problems in the next 6 months. METHODS: Using a cross-sectional research design, data were collected via self-administered questionnaires from convenience samples at a Senior Health Clinic and a Veterans Affairs Hospital outpatient pharmacy. Study subjects were community dwelling adults aged 65 years and older and able to complete the survey in English. Multiple regression analysis was used to identify the significant predictors of intention to use herbal medicines in the TPB within each ethnicity. Independent t tests were used to compare the beliefs underlying the significant predictors of intention with use herbal medicines across the 2 groups. RESULTS: For both Hispanic (N=80) and non-Hispanic (N=171) patients, attitudes toward using herbal medicines was the only significant predictor of the intention to use herbal medicines in the next 6 months (Hispanics, Adj. R(2)=0.59, beta=0.78, P<.001; non-Hispanics, Adj. R(2)=0.57, beta=0.66, P<.001). The magnitude of the beta coefficients did not differ significantly between the 2 ethnicities. Compared with non-Hispanics, Hispanics believed that herbal medicines are cheaper, have fewer side effects, work better, and are more convenient to use than other medicines. CONCLUSIONS: Understanding ethnic differences in behavioral beliefs underlying attitudes toward the use of herbal medicines can help pharmacists and other health care professionals in educating and formulating appropriate counseling strategies specific to older patients of different ethnicities. PMID- 17138513 TI - Demystifying medication safety: making sense of the terminology. AB - BACKGROUND: Although there is a growing interest in medication safety, there remains much confusion about the terminology used to describe the problem. Some have described the classification of medication safety terminology as haphazard. OBJECTIVE: The purpose of this commentary is to help provide some direction by clarifying the terminology. METHODS: A review of the medication safety literature was performed. A description of commonly used terms is provided and the implications of the misuse of terminology are discussed. RESULTS: There are inconsistencies in the definitions of commonly used terms that may affect the accuracy of event rates. This may have an adverse impact on the establishment of medication safety priorities and on the validity of cross-jurisdictional comparisons. CONCLUSIONS: As the medication safety literature continues to expand, it is imperative that standardized terminology be adopted and used consistently. PMID- 17138514 TI - Indications of maturation in research on pharmacist workforce issues. PMID- 17138515 TI - Much needed attention devoted to pharmacy workforce issues. PMID- 17138516 TI - Prediction of pharmacist intention to provide Medicare medication therapy management services using the theory of planned behavior. AB - BACKGROUND: Medicare Part D is a voluntary prescription drug benefit for Medicare beneficiaries. As part of the coverage, medication therapy management services (MTMS) are mandated for beneficiaries with chronic diseases who take multiple medications covered under part D and who are likely to incur annual costs that exceed a specified level. OBJECTIVE: To predict the behavioral intention of pharmacists to provide Medicare medication therapy management services (MTMS) using the theory of planned behavior (TPB) and to determine the relationship between pharmacists' characteristics and intention to provide MTMS. METHODS: The population for this cross-sectional descriptive study consisted of all community pharmacists in Iowa. Data collection occurred through a self-administered anonymous mail survey. Two surveys each were mailed to 500 pharmacies selected through a stratified random sample, 1 survey for the pharmacy manager and 1 survey for a staff pharmacist if applicable. Descriptive statistics and scale reliability were calculated for each of the 4 TPB scales (attitude, subjective norm, perceived behavioral control, and intention). Linear regression was used to predict intent as a function of the other 3 TPB factors, demographic factors, experience, and type of pharmacy. RESULTS: Out of 212 surveys received, 203 had usable data. The usable response rate ranged from 21% to 41%. Pharmacists' intent to provide MTMS was generally positive but varied in strength with a mean score of 22.47 (+/-4.00) and a range of 7-30. Pharmacists mostly agreed that they had appropriate training to provide MTMS but lacked time and support. The linear regression analysis found the constructs of attitude, subjective norm, and perceived behavioral control to be significant predictors of intent (P<.05). Pharmacists with stronger intent to provide MTMS were those who felt they had more control over providing MTMS, felt their peers approved of the provision of MTMS, and had a positive attitude about providing MTMS. Type of pharmacy and pharmacist demographic variables were not significant predictors of intent to provide MTMS. CONCLUSION: Pharmacists showed generally positive intent to provide MTMS. Perceived behavioral control, subjective norm, and attitude were significant predictors of intent (P<.05). Strategies to help pharmacists provide MTMS should focus on finding time and support to provide MTMS rather than individual educational needs. PMID- 17138517 TI - Predicting the impact of Medicare Part D implementation on the pharmacy workforce. AB - BACKGROUND: There is currently a shortage of pharmacist manpower, and it is expected to continue into the near future. It is also likely that the implementation of Medicare Part D will further aggravate the shortage by increasing demand, but it is not clear how much impact it will have. OBJECTIVE: The purpose of this study was to estimate the impact that the new Medicare drug benefit program will have on pharmacy workforce demand. METHODS: Analysis was conducted using forecasting techniques, which combines traditional statistical theory with both quantitative and qualitative methods. The Aggregated Demand Index (ADI) was designated as the dependent variable. A number of independent variables were selected for their potential to affect the workforce, demand for prescriptions or clinical services, and patient population. Data for the identified variables were collected from a variety of sources. Supply and demand data were analyzed at a national level. RESULTS: Both historical and univariate forecasts indicated that the demand for pharmacists will continue to exceed the supply of pharmacists. The ADI ratio of pharmacist demand-to-supply has recently leveled off which means that demand and supply are in an equilibrium that falls to the demand side. Consequently, the Medicare Modernization Act (MMA) is not predicted to produce a dramatic increase in prescription volume, which would change the current demand for pharmacists. Multivariate forecasting models were not robust primarily due to the lack of precise predictor variables. CONCLUSIONS: Despite the reliance on preliminary univariate forecasts and imprecise predictor variables, it appears that the increased use of prescriptions due to the MMA Part D will have minimal impact on pharmacist demand. PMID- 17138518 TI - Prevalence of and interest in unionization among staff pharmacists. AB - BACKGROUND: Although the propensity for staff pharmacists to join a labor union has never been high, conditions in the profession and workplace have changed over the last decade. Some of these changes may result in staff pharmacists joining a labor union, as well as increased interest in staff pharmacists who are currently not union members to join. OBJECTIVES: The objectives of this study were to (1) assess the degree of union membership among staff pharmacists in 6 states, (2) compare the practice settings, work activities and conditions, compensation, and demographic characteristics between union and nonunion staff pharmacists, (3) assess the level of interest in joining a union among nonunion staff pharmacists, and (4) compare the practice settings, work activities and working conditions, wages and benefits, and demographic characteristics between nonunion staff pharmacists interested in joining a union and nonunion staff pharmacists who were not interested in joining a union. METHODS: A biennial pharmacist compensation study was conducted in 6 states (Florida, Iowa, Illinois, Minnesota, Tennessee, Wisconsin) in late 2003. Randomly selected pharmacists were mailed a self administered questionnaire asking about their practice setting, work activities and conditions, wages and benefits, and demographic characteristics. Respondents were also asked to indicate current membership in a union and, if not a member, their desire to unionize their workplace. RESULTS: Compensation and unionization data were provided by 2,180 respondents (27% usable response rate), of which 1,226 (56%) were staff pharmacists. Eight percent of the staff pharmacists were union members, whereas 18% of nonunion members would vote to unionize their workplace. There were few statistically significant differences between union and nonunion staff pharmacists regarding work activities, working conditions, and hourly wages. However, the benefits provided to union staff pharmacists differed from those provided to nonunion staff pharmacists in several ways. Union staff pharmacists were younger than their nonunion counterparts (40.9 vs 44.5 years, P=.01), yet had worked for their current employers a longer time (11.1 vs 7.3 years, P=.03). Nonunion staff pharmacists interested in joining a union differed from those who would not by practice location and setting, working conditions, and benefits. CONCLUSIONS: Although the union membership rate among staff pharmacists is relatively low, there are geographic and practice areas where membership rates are higher. Differences in work activities, working conditions, wages, and benefits were noted between union and nonunion staff pharmacists as well as between those who would join a union and those who would not. These differences merit further investigation, especially with respect to evaluating the effectiveness of unions and identifying factors that may lead nonunionized staff pharmacists to join a union. PMID- 17138519 TI - Market dynamics of community pharmacies in Minnesota. AB - BACKGROUND: From a pharmacist workforce perspective, an understanding of pharmacy location is important for monitoring access points for pharmacist services such as medication dispensing, medication counseling, medication therapy management, and disease management. OBJECTIVE: To understand access to community pharmacies, our goal was to describe changes in pharmacy mix (independent vs chain) between 1992 and 2002 for 87 counties in Minnesota. Study objectives were to describe the association of (1) change in population density, (2) metropolitan designation, change in (3) proportion of nonwhite population, (4) proportion of elderly population, and (5) household income with change in (a) number of community pharmacies overall, (b) number of chain pharmacies, (c) number of independent pharmacies, and (d) the independent-to-total community pharmacy ratio. METHODS: Records from the State of Minnesota Board of Pharmacy were used to examine changes in the community pharmacy mix from 1992 to 2002. Data on county level demographics in 1990 and 2000 were obtained from the US Census Bureau and Datanet. Chi-square analysis was used to test the relationships between the independent and dependent variables. RESULTS: As of 2002, every county in Minnesota had at least one pharmacy as was the case in 1992. The ratio of independent to chain pharmacies changed from approximately 2:1 in 1992 to approximately 1:1 in 2002. Chi-square results revealed that change in population density was significantly associated with change in the number of community pharmacies overall (P<0.001) and with change in chain pharmacies (P=0.03). The findings revealed an interesting u-shaped pattern for the association between population density and change in independent pharmacies. CONCLUSIONS: It appears that chain pharmacies follow changes in population density for making decisions about market entry and exit. The results suggest that some independent pharmacies might be closing due to chain competition in high population density growth areas. However, there was also evidence to suggest that population density decline may be an important determinant of independent pharmacy closures. In such environments, chain pharmacies are not likely to open new pharmacies to replace the independent pharmacy. Such a market dynamic may lead to access problems for citizens of these counties in the future. PMID- 17138520 TI - Assessment of pharmacy manpower and services in West Virginia. AB - BACKGROUND: The shortage of pharmacists across the nation has been much publicized and has been identified as one of the reasons for new schools of pharmacy to open or for existing colleges and schools of pharmacy to increase their class sizes. This article represents the assessment of a new school of pharmacy's evaluation of staffing and practice in its geographic area. OBJECTIVE: This survey represents the first data point within the School of Pharmacy assessment plan and will be repeated at several intervals after the program opens to longitudinally evaluate its impact on pharmacy staffing and services within West Virginia. METHODS: Using a modified Dillman survey methodology a random sample of 548 pharmacists in West Virginia, approximately one-third the active roster, were surveyed regarding staffing and services in West Virginia. RESULTS: A response rate of 32.78% was achieved and findings indicated that there is a staffing shortage of pharmacists within West Virginia, that staffing impacts the services pharmacists provide, and that more pharmacists would be willing to offer disease management services if staffing levels were at appropriate levels. CONCLUSIONS: A shortage of pharmacists does exist in West Virginia and it has implications on patient care. Longitudinal evaluation of the impact of a new pharmacy program will be conducted and the staffing and services within West Virginia should be continued to be studied. PMID- 17138521 TI - Examining gender salary disparities: an analysis of the 2003 multistate salary survey. AB - BACKGROUND: Pharmacist salary and wage surveys have been conducted at the state and national level for more than 20 years; however, it is not known to what extent, if any, wage disparities due to gender still exist. OBJECTIVES: The overall objective of this study was to determine if wage disparities exist among male and female pharmacists at the multistate and individual state level for each of 6 states studied. A secondary objective was to explore the effect of various demographic variables on the hourly wages of pharmacists. METHODS: Data were collected from 1,688 pharmacists in 6 states during 2003 using a cross-sectional descriptive survey design. A multiple regression analysis on hourly wage testing the effects of state of practice, practice setting, position, terminal degree, and years in practice was conducted. Subsequent multiple regression analyses were conducted individually for each of the 6 states to test the effects of the above variables on hourly wage for both male and female pharmacists, followed by state level analyses for male and female pharmacists, respectively. RESULTS: For the pooled data, all variables were found to be significant predictors of hourly wage, except for earning a PharmD degree without a residency or graduate degree. Gender was not a significant predictor of wage disparities in the state-level analyses. Position was the only significant predictor of wage disparities in all states (except Tennessee) such that pharmacists in management positions make significantly higher salaries than those in staff positions. CONCLUSIONS: The results of these analyses suggest that wage disparities due to gender do not exist at the state level for the 6 states surveyed, when controlling for practice setting, position, terminal degree, and years in practice. The larger number of men in management positions may explain lower wages for female pharmacists. PMID- 17138522 TI - Flexible working: understanding the locum pharmacist in Great Britain. AB - BACKGROUND: There is a growing trend in Great Britain (GB) for pharmacists to work as self-employed "locums" rather than as permanent employees. Despite this trend, little is known about their work patterns or why they choose to pursue nonstandard forms of work. OBJECTIVES: The overall aim of the study was to explore why locums choose self-employment over a permanent contract and to explore a number of issues commonly associated with nonstandard working, such as marginalization and job satisfaction. METHODS: A qualitative interview study was undertaken. In-depth telephone interviews were conducted with 34 locum pharmacists randomly selected from the GB register of pharmacists. Locums from a range of age groups, different sectors of practice, and with different work patterns were selected to ensure that a wide range of experiences and views were covered. RESULTS: The need or desire for flexibility was the overriding factor for choosing to work as a locum. A wide range and variety of individual personal circumstances were important drivers, but a desire for work-life balance was fundamental to many. A variety of work patterns were found, ranging from those with more ad hoc working arrangements to those who worked in the same store on a regular basis. Avoiding stress, paperwork, and nonprofessional duties were among reasons for choosing to locum. Disadvantages associated with being a locum included being viewed and treated negatively by peers, and having fewer opportunities for training. No conclusive evidence could be found for locums being marginalized, except for the training issues for some pharmacists. The findings do suggest some cause for concern, with some locums selecting places to work on the basis of attitudes not congruent with socially inclusive approaches to public health care. CONCLUSIONS: The locum workforce is far from homogenous or uniform. Freelance working of this kind has advantages for the individual: freedom and independence. But there may be risks for the profession if nonstandard work practices become more widespread: isolation, lack of social cohesiveness, and amoral attitudes. Improving working conditions and practices may prevent pharmacists from leaving permanent positions. PMID- 17138523 TI - Governance and the pharmaceutical workforce in England. AB - A key component of a commitment to patient safety is ensuring the fitness to practice of health professionals. This article uses the changing landscape of the regulation of the pharmaceutical workforce and the extended roles and responsibilities of pharmacy practitioners in England to explore the challenges for governance within contemporary pharmacy practice, and consider the powers and structures necessary for effective regulation. In England, it is recognised that the approach of the national regulatory body for pharmacy should be consistent with that of the regulators of other health professions, and focus on assuring the competence and performance of practitioners. Pharmacy employers will have complementary adjunct responsibilities. Attention is being concentrated on ensuring the continuing fitness to practice of health professionals, with regular revalidation certain to become a requirement for all. Particular challenges arise in the effective regulation of advanced practitioners, and in ensuring the continuing competence of prescribing practitioners, who in England may come from a range of health disciplines, including pharmacy, and practice in a variety of settings and circumstances. A separate professional register is to be established for pharmacy technicians, whereupon they will also have to demonstrate continuing competence to practice, and be subject to regulation in the same way as pharmacists. Significant change is also planned in the professional control of community pharmacies. In future the law will require a "responsible pharmacist" to be identified for each community pharmacy, rather than insisting on a pharmacist always being present to supervise the dispensing and sale of medicines. PMID- 17138524 TI - Cultural competency: Agenda for Cultural Competency Using Literature and Evidence. AB - BACKGROUND: Cultural competency has been recognized as an important issue relevant to all health professions. A research agenda is needed to establish a systematic approach to developing an understanding of factors relevant to the delivery of culturally competent health care. OBJECTIVE: Within the context of existing literature, evidence-based, concrete recommendations are developed as an Agenda for Cultural Competency Using Literature and Evidence (ACCULTURE). METHODS: First, key points representing opportunities for intervening in promotion of cultural competent health care are discussed. Following is a review of existing literature with a focus on identifying next steps for future research. Recommendations for licensing, education, and continuing education requirements suggest developing educational research establishing course content and delivery strategies that have measurable impact on improving cultural competency. In addition, existing initiatives need to be evaluated regarding effectiveness in recruiting, retaining, and preparing a diverse workforce. Patient care recommendations focus on further developing an understanding of the factors impacting health outcomes for culturally diverse patients. RESULTS: Further work is needed for translating theoretically-based research into concrete curricula maintaining evidence-based outcomes. It is important to continue with promoting policies ensuring that research and clinical trials include diverse samples and a broad range of variables implicated in differential outcomes. CONCLUSIONS: Based on connections between cultural competency and workforce diversity established within existing literature, data are needed regarding the effectiveness of existing initiatives promoting scholarships, grants, and incentives for improving workforce diversity and funding research on diversity issues. Finally, additional research is needed to evaluate existing and new policies for funding services and access for health services. PMID- 17138525 TI - Learning to minimize energy costs and maximize mechanical work in a bimanual coordination task. AB - The authors addressed the hypothesis that economy in motor coordination is a learning phenomenon realized by both reduced energy cost for a given workload and more external work at the same prepractice metabolic and attentional energy expenditure. "Self-optimization" of movement parameters has been proposed to reflect learned motor adaptations that minimize energy costs. Twelve men aged 22.3 -/+ 3.9 years practiced a 90 degrees relative phase, upper limb, independent ergometer cycling task at 60 rpm, followed by a transfer test of unpracticed (45 and 75 rpm) and self-paced cadences. Performance in all conditions was initially unstable, inaccurate, and relatively high in both metabolic and attentional energy costs. With practice, coordinative stability increased, more work was performed for the same metabolic and attentional costs, and the same work was done at a reduced energy cost. Self-paced cycling was initially below the metabolically optimal, but following practice at 60 rpm was closer to optimal cadence. Given the many behavioral options of the motor system in meeting a variety of everyday movement task goals, optimal metabolic and attentional energy criteria may provide a solution to the problem of selecting the most adaptive coordination and control parameters. PMID- 17138526 TI - Perceived reachability in single- and multiple-degree-of-freedom workspaces. AB - In comparisons of perceived (imagined) and actual reaches, investigators consistently find a tendency to overestimate. A primary explanation for that phenomenon is that individuals reach as a "whole-body engagement" involving multiple degrees of freedom (m-df). The authors examined right-handers (N = 28) in 1-df and m-df workspaces by having them judge the reachability of targets at midline, right, and left visual fields. Response profiles were similar for total error. Both conditions reflected an overestimation bias, although the bias was significantly greater in the m-df condition. Midline responses differed (greater overestimation) from those of right and left visual fields, which were similar. Although the authors would have predicted better performance in the m-df condition, it seems plausible that if individuals think in terms of m-df, they may feel more confident in that condition and thereby exhibit greater overestimation. Furthermore, the authors speculate that the reduced bias at the side fields may be attributed to a more conservative strategy based in part on perceived reach constraints. PMID- 17138527 TI - Gender-specific movement strategies using a computer-pointing task. AB - Females typically demonstrate a movement time advantage for tasks requiring high levels of manual dexterity, whereas males are notably better at targeting activities. According to D. Kimura (2000), the hunter-gatherer hypothesis primarily accounts for those performance advantages; that dichotomy fails, however, when one makes movement outcome predictions for tasks that are not clearly fine-motor or interceptive in nature. Investigators have recently proposed that time constraints (M. Peters, 2005) and gender-specific response style differences (M. Peters & P. Campagnaro, 1996; L. E. Rohr, 2006) affect motor performance. Here, the author used a computer-pointing task measuring both movement error and movement time in 16 participants to further investigate response style differences. Kinematic and linear regression analyses between resultant error and both movement time and task difficulty reinforced the notion that gender-specific movement biases emphasize speed and accuracy, respectively, for men and women. PMID- 17138528 TI - Integration of intermittent visual samples over time and between the eyes. AB - The authors investigated the integration of alternate disparate monocular inputs for binocular perception in 1-handed catching experiments (N = 14, 32, 22, and 15 participants, respectively in Experiments 1-4). They varied the no-vision interval between alternate monocular samples to measure catching performance, and they compared the alternating monocular conditions with binocular and monocular conditions with equal no-vision intervals. They found no evidence of a binocular advantage for one-handed catching in the alternating monocular conditions. Performance in monocular and alternating monocular conditions did not differ across no-vision intervals ranging from 0-80 ms and was particularly worse than performance in binocular viewing conditions when the no-vision interval was 40 ms or more. The authors argue that the dissimilarity between disparate monocular inputs created by the approaching object limited the integration of those inputs and subsequent binocular perception. PMID- 17138529 TI - Information entropy and the variability of space-time movement error. AB - The authors investigated the effects of movement time and movement distance on the information entropy and variability of spatial and temporal error in a discrete aiming movement. In Experiment 1, the authors held movement distance (100 mm) constant and manipulated 11 movement times (300-800 ms) of 8 participants. In Experiment 2, the authors tested 6 movement distances at 2 given movement times (15-60 mm at 300 ms; 40-240 mm at 800 ms) in 8 participants. The variability and entropy for spatial error increased with average movement velocity, whereas the variability and entropy for temporal error decreased as a function of average movement velocity. The common variance between variable error and entropy averaged about 84% and 72% for spatial and temporal errors, respectively, suggesting that the probabilistic approach of entropy reveals features that are not present in the standard deviation index of variability. The findings provide further evidence that information entropy may be a useful single index representation of variability in the movement speed-accuracy relation. PMID- 17138530 TI - Reprogramming of interceptive actions: time course of temporal corrections for unexpected target velocity change. AB - The authors investigated the time course of reprogramming of the temporal dimension of motor acts in a task requiring interception of a moving target. The target moved at a constant velocity on a monitor screen; in part of the trials, target velocity was unexpectedly increased or decreased. Those modifications were produced at different moments during target displacement, leaving periods of time from 100 to 800 ms for movement timing correction. The authors assessed the effects of probability of target velocity change (25% vs. 50%), uncertainty about direction of velocity change (unidirectional vs. bidirectional), and direction of velocity change (increase vs. decrease). Analysis of 24 participants' arm acceleration showed that fast adjustments took place between 100 and 200 ms after target velocity change similarly for all uncertainty conditions. Analysis of temporal error indicated that the combination of high probability of target velocity change and certainty on direction of target velocity change led to the most successful movement timing reprogramming. For the other experimental conditions, temporal accuracy was still poor when a period of 800 ms was available for correction. Movement reprogramming was a continuous process that was more efficient for target velocity increase than for target velocity decrease. PMID- 17138531 TI - Numerosity and rhythmicity in stimulus-response compatibility. AB - When people must respond discriminatively to 1 or 2 stimuli by making 1 or 2 taps of a response key, they initiate the response more rapidly when the correct number of taps matches the number of stimuli (compatible condition) than when it mismatches (incompatible condition; J. O. Miller, S. G. Atkins, & F. Van Nes, 2005). Miller et al. sometimes found an effect of compatibility on response execution time, as reflected in the interresponse intervals between successive taps. The authors report 2 further experiments (N = 8 participants) in which they generalized the numerosity compatibility effects on response-initiation time and interresponse intervals to 2- versus 3-stimulus sequences. In addition, they varied gap length between stimuli to see whether the rhythm of the stimulus would influence that of the response. Weak rhythmicity effects were repeatedly found, but those were too small to suggest a plausible alternative explanation for the numerosity compatibility effect on response-initiation time. PMID- 17138533 TI - [Polymorphism analysis of seven Y-STR loci in Shui population in Guizhou, China]. AB - The objective of this study was to investigate the polymorphism of seven Y specific STR loci in Shui ethnic population of Guizhou, China, and to obtain the polymorphism information in this minority. One trinucleotide STR locus and six tetranucleotide STR loci were simultaneously amplified with fluorescently labeled primers, and genotypes were determined with ABI PRISM 377 DNA Sequencer. Allele frequencies, genetic diversity and haplotype diversity were calculated. Among 94 unrelated males, 6, 4, 6, 2, 3, 5, 4 alleles were observed in loci DYS19, DYS389 I, DYS389 II, DYS390, DYS391, DYS392 and DYS393, respectively. Altogether, 27 haplotypes were identified for the seven Y-STR loci. The genetic diversity values for each locus ranged from 0.124 (DYS389 I) to 0.630 (DYS19). The haplotype diversity value was 0.868. High haplotype diversities were found in Shui population of Guizhou. The study suggests that these seven Y-STR loci are valuable Y-specific markers for establishing a Y-STR database, understanding ethical origin and migrations and for personal identification. PMID- 17138532 TI - [Mapping of gene underlying autosomal dominant non-syndromic hearing loss(DFNA)]. AB - Hereditary non-syndromic sensorineural hearing loss is a genetically highly heterogeneous group of disorders. To date, at least 50 loci for autosomal dominant non-syndromic sensorineural hearing loss (DFNA) have been identified by linkage analysis. Here we report a huge family with late onset autosomal dominant hereditary non-syndromic hearing loss. In this family, 73 of 170 family members have been conducted physical examination, pure-tone audiometry, immittance testing and auditory brainstem response testing (ABR). The results indicated that 39 of 73 tested family members have sensorineural hearing loss in various degrees. No associated visible abnormalities in other systems were found in this family. After exclusion of the 14 known DFNA loci with markers from the Hereditary Hearing Loss Homepage (URL: http://dnalab-www.uia.ac.be/dnalab/hhh), a genome wide scan was carried out using 382 highly informative microsatellite markers at approximately 9.2 cM intervals throughout the genome. Linkage analysis was carried out under a fully penetrant autosomal dominant mode of inheritance with no phenocopies. A maximum two-point LOD score of 6.69 at theta=0 was obtained for marker D14S1040. Haplotype analysis placed the locus within a 7.6 cM genetic interval defined by marker D14S1021 and D14S70, overlapping with the DFNA9 locus. PMID- 17138534 TI - [The association of MMP-13 polymorphism with susceptibility to esophageal squamous cell carcinoma and gastric cardiac adenocarcinoma]. AB - Matrix metalloproteinase-13 (MMP-13) belongs to a family of enzymes that degrade all components of the extracellular matrix. Polymorphism in the promoter region of the MMP-13 gene may modify its transcriptional activity and protein level. This study was designed to investigate the correlation of MMP-13 A-77G single nucleotide polymorphism (SNP) with susceptibility to esophageal squamous cell carcinoma (ESCC) and gastric cardiac carcinoma (GCA) in a population from a high incidence region of Hebei province. MMP-13 promoter SNP (A-77G) was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis in 316 ESCC patients, 243 GCA patients and 609 healthy controls. The overall genotype and allelotype distributions of MMP-13 in ESCC and GCA patients were not significantly different from those in healthy controls (P>0.05). When stratified for smoking status, the A/A, A/G, G/G genotype frequencies of MMP-13 in GCA patients and smoker controls group were 27.1%, 44.1%, 28.8% and 17.8%, 60.5%, 21.7% respectively. There was a significant difference between the two groups (chi2=9.01, P=0.01). The A/G genotype frequency in GCA patients was significantly lower than that in the controls, when compared to the A/A genotype, suggesting that individuals with the A/G genotype may have reduced susceptibility to GCA in the smoker group (OR=0.48, 95% CI=0.280.83). However, the G/G genotype showed no significant influence on the risk of developing GCA. Thus, in the region of Hebei Province where there is a high incidence of GCA and ESCC, the MMP 13 A-77G SNP may not be associated with cancer susceptibility. The A/G genotype may confer protection against GCA for smokers. PMID- 17138535 TI - [Correlation analysis of microsatellite DNA markers with wool traits in Liangshan semi-fine wool sheep]. AB - Eighteen microsatellites on chromosomes 1, 2, 3, 9 were studied to detect their genotypes in 206 individuals in a Liangshan Semi-fine Wool Sheep nucleus breeding population. A GLM procedure was used to analyze the effects of these 18 microsatellites on wool traits. Results uncovered 7 loci that had a significant impact on wool traits. Genotypes of that favorably affect wool traits were determined. PMID- 17138536 TI - [Nuclear transfer of goat somatic cells transgenic for human lactoferrin]. AB - Transgenic animal mammary gland bioreactors are being used to produce recombinant proteins with appropriate post-translational modifications, and nuclear transfer of transgenic somatic cells is a more powerful method to produce mammary gland bioreactor. Here we describe efficient gene transfer and nuclear transfer in goat somatic cells. Gene targeting vector pGBC2LF was constructed by cloning human lactoferrin (LF) gene cDNA into exon 2 of the milk goat beta-casein gene, and the endogenous start condon was replaced by that of human LF gene. Goat fetal fibroblasts were transfected with linearized pGBC2LF and 14 cell lines were positive according to PCR and Southern blot. The transgenic cells were used as donor cells of nuclear transfer, and some of reconstructed embryos could develop to blastocyst in vitro. PMID- 17138537 TI - [Genetic variation in the 5' flanking region of bovine TLR4 gene and correlation with mastitis]. AB - Toll-like receptor 4 recognizes pathogen ligands and mediates signaling to initiate innate and adaptive immune responses. In this experiment, a 477 bp segment of the 5'-flanking region of TLR4 gene of Chinese Holstein, Sanhe cattle and Chinese simmental was amplified by polymerase chain reaction. After sequencing, a polymorphic site in amplified production of TLR4 was identified of having either a G or a C at position 245. This polymorphism in the three populations was detected by digesting the fragment with restriction endonuclease Msp I. Results showed that both alleles (A and B) were found in the three populations and the value of polymorphism information content indicated that this was a moderate polymorphism. Chi2 test indicated that the polymorphism locus in Sanhe cattle did not fit Hardy-Weinberg equilibrium (P< 0.05). In addition, the effect of the TLR4 polymorphism on somatic cell score was analyzed, and the results indicated that the somatic cell score were significantly affected by lactation month and the type of breeds(P<0.05), but not by different genotypes (P > 0.05). PMID- 17138538 TI - [Relationship between microsatellite DNA polymorphism and hind-quarter development in sheep chromosome 18]. AB - Ten highly polymorphic microsatellite loci possibly linked to or correlated with the Callipyge gene were selected, according to the genetic map and linkage map of sheep chromosome 18. They were ILSTS54, TGLA337, HH47, TGLA122, BP33, OB2, BM3413, MCM38, MCMA26 and CSSM18. Polymorphisms of these microsatellites were detected in 61 Dorset (male) x Xinjiang fine wool sheep (female) samples and 76 Suffolk (male) x Xinjiang fine wool sheep (female) samples. Results showed that the number of alleles for the 10 microsatellite loci, heterozygosity and PIC (polymorphism information content) in the Dorset population were 8-16, 0.8370 0.9252, and 0.8221-0.9167, respectively. The same parameters in the Suffolk population were 5-10, 0.7603-0.8913 and 0.7176-0.8809, respectively. The effect of these loci on hindquarter width was analyzed in a generalized linear model. Results showed that, in the Dorset group, BM3413, MCMA26 and CSSM18 each had a significant effect on hindquarter width (P<0.05), while the other seven loci had no effect on it (P>0.05). In the Suffolk group, TGLA122, BM3413, MCM38 and CSSM18 had a significant effect on hindquarter width (P<0.05), while the other six loci did not (P>0.05). Our results also indicated that the cause of hindquarters hypertrophy in Xinjiang meat sheep may be different from the A-to-G mutation between the region of DLK1 and GTL2 . There may be other genes or QTL (quantitative trait loci) that affect hindquarter muscle development on chromosome 18 in Xinjiang meat sheep. PMID- 17138539 TI - [Cloning, sequencing and prokaryotic expression of cDNAs for the antifreeze protein family from the beetle Tenebrio molitor]. AB - The partial cDNA sequence coding for the antifreeze proteins in the Tenebrio molitor was obtained by RT-PCR. Sequence analysis revealed nine putative cDNAs with a high degree of homology to Tenebrio molitor antifreeze proteins. The recombinant pGEX-4T-1-tmafp-XJ430 was introduced into E. coli BL21 to induce a GST fusion protein by IPTG. SDS-PAGE of the fusion protein demonstrated that the antifreeze protein migrated at a size of 38 kDa. The immunization was performed by intra-muscular injection of pCDNA3-tmafp-XJ430, and then antiserum was detected by ELISA. The titer of the antibody was 1:2,000. Western blotting analysis showed the antiserum was specific against the antifreeze protein. This finding could lead to further investigation of the properties and function of antifreeze proteins. PMID- 17138540 TI - [Population genetic variation and structure analysis on five populations of mirror carp Cyprinus carpio L. using microsatellites]. AB - In this paper, population genetic variability and genetic structure of five populations of an important cultivation species, mirror carp (Cyprinus carpio L.) were analyzed using 30 microsatellite loci. The observed (Ho) and expected (He) heterozygosity values, polymorphic information content (PIC) and number of effective alleles (Ae) were all determined. The genetic similarity coefficient and Nei's standard genetic distance were computed based on the allele frequencies. The Hardy-Weinberg equilibrium was checked by chi2 test. Genetic differentiation and hierarchical partition of genetic diversity were evaluated by FST and Nm. A dendrogram was constructed based on UPGMA methods using PHYLIP software package supported by a bootstrap value of 91.0%. Totally 7,083 fragments were procured. Their lengths were from 102 bp to 446 bp. For each locus, 1-16 alleles were amplified, adding up to 356 alleles in all the 5 populations. We found the genetic variability level was relatively high in all five populations, as shown by Ae = 1.07-2.30, He= 0.70-0.78 and PIC=0.69-0.75, respectively. The genetic similarity coefficients were all above 0.52, indicating their close genetic relationships. The UPGMA phylogenetic tree showed mirror carps sampled from Donggang, Fengcheng and Liaozhong were clustered into one group and the other two populations, both collected from Songpu, were grouped together. There were obvious relations between genetic distances and geographical distributions of the five populations. No fragments were amplified from some loci of EST-SSRs, which may suggest the loss of these loci in mirror carp genome or sequence divergence at the primer binding sites. These null alleles may result from selection because functional genes are under more selection pressure than non encoding loci. Overall, population genetic variation is high for each of the five mirror carp, and the differentiations are also significant among populations. PMID- 17138541 TI - [Analysis of genetic variation of abalone (Haliotis discus hannai) populations with microsatellite markers]. AB - Microsatellite markers were used to access the genetic variation in three populations of abalone Haliotis discus hannai. Two wild populations were collected from the sea areas in Changdao, Shandong and Dalian, Liaoning respectively. A cultivated population originated from the sea area in Kongdongdao, Shangdong. Six microsatellite loci were screened for genetic polymorphism. Polymorphic information content (PIC) value per loci was greater than 0.5 and can be used to analysis of genetic structure of the three abalone populations. Fifty-seven alleles were amplified from the three populations in six microsatellite loci. The average number of alleles (A) was 9.50 and the effective number of alleles (Ne) was 5.8572. The mean observed heterozygosity (Ho) and the mean expected heterozygosity (He) were 0.6925 and 0.7966, respectively. The Ho and He of two wild abalone populations were higher than that of cultured population. All these results provide a basis for conservation and utilization of genetic diversity of Haliotis discus hannai. PMID- 17138542 TI - [Amplification of Ds flanking sequences from rice genomic DNA of hybrids of Ac x Ds lines and the analysis of Ds insertions]. AB - Ac and Ds insertions among the genomic DNAs of hybrids of Ac x Ds lines were screened by PCR. The genomic DNAs, which were proved to harbour both Ac and Ds, were used as templates in TAIL-PCR to clone the Ds flanking sequences. The cloned specific fragments were sequenced, and the sequenced Ds flanking sequences were used as query sequences to perform on-line sequence comparing analysis against GenBank by employing BLAST program of NCBI. The information about the chromosome location of Ds-inserted genes, or genes immediately downstream of the inserted sites, and their functional innotations were achieved. Based on the analysis from the cloned 93 Ds-flanking sequences, it was found that 21 hybrid plants had Ds insertions in genic regions, whereas the remaining 72 samples's intergenic regions were inserted by Ds element. Moreover, among the 72 regions, 12 were inserted immediately upstream (within 3 kb) of specific genes. Also, the strategies to improve the performance in cloning the Ds flanking sequences and in screening the Ac/Ds lines were emphasized. PMID- 17138543 TI - [Genetic analysis and SSR marker of tolerance to submergence in rice]. AB - Submergence stress has severe harm to rice (Oryza sativa L.) production. It is important to identify molecular markers associated with tolerance to submergence for marker-assisted selection (MAS). F1 and F2 populations ware obtained from reciprocal crosses between submergence tolerant parent FR13A and submergence sensitive parent IR39595-503-2-1-2. No difference in reciprocal crosses in F1 population was detected for tolerance to submergence. This indicates that the submergence character is controlled by nucleic factor(s). Tolerance to submergence derived from FR13A appeared to be a quantitative-qualitative trait as revealed by the segregation of F2 population in two submergent experiments. Under light submergence stress, multiple minor genes might work and the trait inherited quantitatively it appeared a quantity trait, While under severe submergence stress, it exhibited a qualitative inheritance controlled by a major gene. Among the 187 pairs of SSR primers, 73 (39%) were polymorphic between the two parents and were used to tag submergence in the F2 population. Primer RM219 was detected to be linked to submergence tolerant trait loci. This result confirmed that tolerance to submergence is controlled by the major gene sub1, which will be helpful in improvement of the trait in rice. PMID- 17138544 TI - [The mixed major gene plus polygenes inheritance for female fertility in wheat (Triticum aestivum L.)]. AB - Three sets of data for the P1, P2, F1, and F2 populations derived from three crosses between the normal fertility wheat (Triticum aestivum L.) cultivars with different ecotypes and the female sterile line (XND126) were used to investigate the inheritance of female fertility in wheat using mixed major gene plus polygenes inheritance model in 2005 and 2006. The results from the joint segregation analysis of the four generations showed that female fertility in wheat is controlled by two major genes plus polygenes, and the interaction between the two major genes is also detected. PMID- 17138545 TI - [Development of Dasypyrum genome specific marker by using wheat microsatellites]. AB - One hundred and two SSR primer pairs, distributed in chromosome 1A to 7A, 1B to 7B, 1D to 7D of Triticum aestivum, were investigated on Dasypyrum breviaristatum, D.villosum, wheat-Dasypyrum amphiploids and its derivatives, with the control of common wheat Chinese Spring and elite wheat cultivars. A specific polymorphic DNA fragment of about 400 bp (415 bp-long by sequenced, named Xgwm301/415) amplified by primer pair Xgwm301 was obtained in all lines containing Dasypyrum chromosomes, but there were not the case in the tested common wheat. Furthermore, PCR analysis was performed on a set of T. aestivum-Dasypyrum addition, the result showed that all the seven pairs of villosum chromosomes contain Xgwm301/415. Therefore, Xgwm301/415 is a genome-specific polymorphic DNA segment for genera of Dasypyrum, and it could be used as a molecular marker for detection of chromosomes of Dasypyrum in wheat. PMID- 17138546 TI - [Expression of rabbit neutrophile peptide-1 in nitrate reductase-deficient mutant of Chlorella ellipsoidea]. AB - Application of transgenic Cholrella as bioreactor to express rabbit neutrophile pepetide-1 (NP-1) shows great practical value. In this paper, an NP-1 expression vector containing two selective marker genes NPTII and nitrate reductase gene was constructed. The NP-1 gene was transformed into the nitrate reductase-deficient mutant nrm-4 of Chlorella ellipsoidea via electroporation, and the transgenic alga expressed the active NP-1 were obtained. PMID- 17138547 TI - [Research advances on the relationship of PI3-kinase/Akt/mTOR pathway and epigenetic modification]. AB - The mammalian target of rapamycin (mTOR) has been shown to link growth factor signaling and posttranscriptional control of protein translation through activation of the PI3K/Akt pathway, which is frequently involved in cell cycle progression. The inhibition of mTOR has promising potential in anticancer and immunosuppressive therapies, and additional phase II clinical trials are ongoing. Epigenetic modification, which involves DNA methylation, histone modification and chromatin remodeling, as well as the recently described RNAi mechanism, can initiate the formation of silenced chromatin. Persistent activation or inhibition of the mTOR pathway may affect epigenetic modification. In this paper, we reviewed the research advances in the relationship between PI3K/Akt/mTOR and epigenetic modification. PMID- 17138548 TI - [The ubiquitin-proteasome proteolytic pathway and tumorigenesis]. AB - The ubiquitin-proteasome proteolytic pathway, a major pathway for protein degradation in cells, plays a critical role in the protein metabolism. So abnormality of the ubiquitin-proteasome proteolytic pathway is closely related to many diseases, especially cancer. In this paper, we reviewed the study of the significant role of the ubiquitin-proteasome proteolytic pathways during the cell cycle, DNA repair and apoptosis, especially the relationship between these pathways and tumorigenesis. PMID- 17138549 TI - [Advances in research of the structure and function of plant centromeres]. AB - Centromeres are the chromosomal domains necessary for faithful chromosome segregation and transmission during mitosis and meiosis in eukaryotes. In the last decade, centromeres in some plant species including Arabidopsis, rice and maize have been deeply studied at molecular level. Centromeric DNAs evolve rapidly and are little conserved among various plants, but the types of centromeric DNA sequences and their organization patterns within centromeres are basically similar in plants. Plant centromeres are usually composed of clusters of tandemly arrayed satellite repeats that are interspersed with centromere specific retrotransposons. In contrast to centromeric DNA, structural and transient centromeric/kinetochoric proteins are conserved among eukaryotes including plants. As the cases in other eukaryotes, the presence of CENH3 (centromeric histone H3)-containing nucleosomes is the fundamental feature of plant functional centromeres, and CENH3 plays critical roles in the identity and maintenance of plant centromeric chromatin. PMID- 17138550 TI - [The structure and function of plant WRKY transcription factors]. AB - WRKY transcription factors that are unique to plants are the new type transcriptional regulatory factors in which N-terminal ends contain a conserved WRKYGQR amino acids sequences. WRKY transcription factors regulate the target genes expression that contain the W-box elements in the promoter regions by specifically binding to (T)(T)TGAC(C/T) sequence. Therefore, the WRKY transcription factors participate in the plant various kinds defense responses and regulate the plant growth and development. This article reviews the progress of the basic structure and biological function of plant WRKY transcription factors. PMID- 17138551 TI - [Advance in quantitative traits from phenotypic variation to gene discovery]. AB - With the rapid development of molecular biotechnology, many improved approaches have been provided for studying on the genetic bases of quantitative traits. Many studies paid attention to dissect the genetic bases using different emendatory approaches from phenotypic variation to gene discovery, mainly containing QTL mapping, chemical mutagenesis etc. Especially, QTL analysis was executed for a lot of important quantitative traits. This paper summarizes the development of quantitative genetics, the recent progress on QTL mapping populations and methods, the status of QTL locating and QTG identification, positional cloning, and QTL application in breeding. PMID- 17138552 TI - [Progress on Agrobacterium-mediated transformation of banana]. AB - The development of improved Musa by conventional breeding remains to be a difficult endeavor because of the long generation times, various levels of ploidy, sterility of most edible cultivars and limited genetic variability. Therefore, genetic engineering may offer an alternative method for crop enhancement. The transformation system mediated by Agrobacterium tumefaciens has been used successfully to transform banana during the past 10 years. This article reviews recent progresses on transformation of banana mediated by Agrobacterium tumefaciens and summarizes the factors affecting transformation efficiency. Existing problems and its prospects are briefly discussed. PMID- 17138553 TI - [The application of large-scale yeast two-hybrid to study protein--protein interaction]. AB - The principle of yeast two-hybrid were summarized, the basis for large scale yeast two-hybrid screening of protein-protein interaction was given and the main application of yeast two-hybrid in large scale screening of protein-protein interaction was presented. The problems of large scale yeast two-hybrid screening of protein-protein interaction were also discussed. Because there were a lot of false positives and false negatives in the result of protein-protein interaction obtained by large-scale yeast two-hybrid screening, the employment of other methods to study protein-protein interaction in large scale in parallel was proposed if possible. PMID- 17138554 TI - [Advances in plant proteomics. II. Application of proteome techniques to plant biology research]. AB - Proteome techniques have widely been applied to the fields of plant genetics, plant development, and plant physiology and ecology to investigate plant genetic diversity, plant development such as seed maturation and germination processes, differentiation of plant tissue and organ, separation and functional identification of novel component of various organells, mechanisms of plant adapted to abiotic or biotic stresses including high temperature, low temperature, high salt, drought, and pathogens and insects, and interaction of plant with microbe. The prospects of plant proteomics are discussed. PMID- 17138555 TI - [Progress in the study on alternative splicing and functions of kininogen genes]. AB - The kininogen gene and its coded proteins are slightly different in various species. In human beings, bovine and mouse, for example, there is an alternatively spliced kininogen gene K, encoding two kinds of proteins. By contrast, there is still another constituted spliced kininogen gene T which encodes only one kind of protein in the rat. The kininogen, belonging to the family 3 of cystatin superfamily, is a kind of multifunctional proteins with multiple domains, which maintains the normal physiological condition in human and some other organisms. The antagonism of hemoglutination and antihemoglutination of kininogen can not only recuperate the damaged blood vessels to prevent them from bleeding ceaselessly, but also restrain the formation of thrombus. In this review, we briefly discussed alternative splicing of kininogen gene and the multifunction of kininogen protein, as well as primarily hemoglutination and antihemoglutination, in human beings, bovine, mouse and rat which have been currently studied in detail. Our aims are to provide the beneficial references for further understanding the mechanism of evolution and alternative splicing of kininogen gene, and elucidating the multifunction roles of kininogen protein. Besides, it would be helpful for developing new medicines to regulate the vascular permeability and blood pressure and to restrain tumor. PMID- 17138556 TI - Crystal structure of human inosine triphosphatase. Substrate binding and implication of the inosine triphosphatase deficiency mutation P32T. AB - Inosine triphosphatase (ITPA) is a ubiquitous key regulator of cellular non canonical nucleotide levels. It breaks down inosine and xanthine nucleotides generated by deamination of purine bases. Its enzymatic action prevents accumulation of ITP and reduces the risk of incorporation of potentially mutagenic inosine nucleotides into nucleic acids. Here we describe the crystal structure of human ITPA in complex with its prime substrate ITP, as well as the apoenzyme at 2.8 and 1.1A, respectively. These structures show for the first time the site of substrate and Mg2+ coordination as well as the conformational changes accompanying substrate binding in this class of enzymes. Enzyme substrate interactions induce an extensive closure of the nucleotide binding grove, resulting in tight interactions with the base that explain the high substrate specificity of ITPA for inosine and xanthine over the canonical nucleotides. One of the dimer contact sites is made up by a loop that is involved in coordinating the metal ion in the active site. We predict that the ITPA deficiency mutation P32T leads to a shift of this loop that results in a disturbed affinity for nucleotides and/or a reduced catalytic activity in both monomers of the physiological dimer. PMID- 17138557 TI - Glucose stimulates Ca2+ influx and insulin secretion in 2-week-old beta-cells lacking ATP-sensitive K+ channels. AB - In adult beta-cells glucose-induced insulin secretion involves two mechanisms (a) a K(ATP) channel-dependent Ca(2+) influx and rise of cytosolic [Ca(2+)](c) and (b) a K(ATP) channel-independent amplification of secretion without further increase of [Ca(2+)](c). Mice lacking the high affinity sulfonylurea receptor (Sur1KO), and thus K(ATP) channels, have been developed as a model of congenital hyperinsulinism. Here, we compared [Ca(2+)](c) and insulin secretion in overnight cultured islets from 2-week-old normal and Sur1KO mice. Control islets proved functionally mature: the magnitude and biphasic kinetics of [Ca(2+)](c) and insulin secretion changes induced by glucose, and operation of the amplifying pathway, were similar to adult islets. Sur1KO islets perifused with 1 mm glucose showed elevation of both basal [Ca(2+)](c) and insulin secretion. Stimulation with 15 mm glucose produced a transient drop of [Ca(2+)](c) followed by an overshoot and a sustained elevation, accompanied by a monophasic, 6-fold increase in insulin secretion. Glucose also increased insulin secretion when [Ca(2+)](c) was clamped by KCl. When Sur1KO islets were cultured in 5 instead of 10 mm glucose, [Ca(2+)](c) and insulin secretion were unexpectedly low in 1 mm glucose and increased following a biphasic time course upon stimulation by 15 mm glucose. This K(ATP) channel-independent first phase [Ca(2+)](c) rise was attributed to a Na(+)-, Cl(-)-, and Na(+)-pump-independent depolarization of beta-cells, leading to Ca(2+) influx through voltage-dependent calcium channels. Glucose indeed depolarized Sur1KO islets under these conditions. It is suggested that unidentified potassium channels are sensitive to glucose and subserve the acute and long-term metabolic control of [Ca(2+)](c) in beta-cells without functional K(ATP) channels. PMID- 17138558 TI - Translational control of glial glutamate transporter EAAT2 expression. AB - Glutamate is the major excitatory neurotransmitter in the central nervous system. Its activity is carefully modulated in the synaptic cleft by glutamate transporters. The glial glutamate transporter EAAT2 is the main mediator of glutamate clearance. Reduced EAAT2 function could lead to accumulation of extracellular glutamate, resulting in a form of cell death known as excitotoxicity. In amyotrophic lateral sclerosis and Alzheimer disease, EAAT2 protein levels are significantly decreased in affected areas. EAAT2 mRNA levels, however, remain constant, indicating that alterations in EAAT2 expression are due to disturbances at the post-transcriptional level. In the present study, we found that some EAAT2 transcripts contained 5'-untranslated regions (5'-UTRs) greater than 300 nucleotides. The mRNAs that bear long 5'-UTRs are often regulated at the translational level. We tested this possibility initially in a primary astrocyte line that constantly expressed an EAAT2 transcript containing the 565-nt 5'-UTR and found that translation of this transcript was regulated by many extracellular factors, including corticosterone and retinol. Moreover, many disease-associated insults affected the efficiency of translation of this transcript. Importantly, this translational regulation of EAAT2 occurred in vivo (i.e. both in primary cortical neurons-astrocytes mixed cultures and in mice). These results indicate that expression of EAAT2 protein is highly regulated at the translational level and also suggest that translational regulation may play an important role in the differential EAAT2 protein expression under normal and disease conditions. PMID- 17138559 TI - Probing the architecture of an L-type calcium channel with a charged phenylalkylamine: evidence for a widely open pore and drug trapping. AB - Voltage-gated calcium channels are in a closed conformation at rest and open temporarily when the membrane is depolarized. To gain insight into the molecular architecture of Ca(v)1.2, we probed the closed and open conformations with the charged phenylalkylamine (-)devapamil ((-)qD888). To elucidate the access pathway of (-)D888 to its binding pocket from the intracellular side, we used mutations replacing a highly conserved Ile-781 by threonine/proline in the pore-lining segment IIS6 of Ca(v)1.2 (1). The shifted channel gating of these mutants (by 30 40 mV in the hyperpolarizing direction) enabled us to evoke currents with identical kinetics at different potentials and thus investigate the effect of the membrane potentials on the drug access per se. We show here that under these conditions the development of channel block by (-)qD888 is not affected by the transmembrane voltage. Recovery from block at rest was, however, accelerated at more hyperpolarized voltages. These findings support the conclusion that Ca(v)1.2 must be opening widely to enable free access of the charged (-)D888 molecule to its binding site, whereas drug dissociation from the closed channel conformation is restricted by bulky channel gates. The functional data indicating a location of a trapped (-)D888 molecule close to the central pore region are supported by a homology model illustrating that the closed Ca(v)1.2 is able to accommodate a large cation such as (-)D888. PMID- 17138560 TI - Interactions of human O6-alkylguanine-DNA alkyltransferase (AGT) with short single-stranded DNAs. AB - The O6-alkylguanine-DNA alkyltransferase (AGT) repairs O6-alkylguanine and O4 alkylthymine adducts in single-stranded and duplex DNAs. Here we characterize the binding of AGT to single-stranded DNAs ranging in length from 5 to 78 nucleotides (nt). Binding is moderately cooperative (37.9 +/- 3.0 55 mm Hg. CONCLUSIONS: Patients with head injury appear to have a near normal distribution of blood pressure readings that are skewed towards higher values. The relationship between BP and ICP may be triphasic. PMID- 17138595 TI - Dynamics of oscillatory activity during auditory decision making. AB - Perceptual decision making requires the comparison and integration of sensory evidence to generate a behavioral response. We used magnetoencephalography to investigate the temporal dynamics of decision making during an auditory task that required forced-choice decisions about whether a pair of syllables S1 and S2 differed either in their acoustic patterns or in the perceived position of their sound sources. Conditions with easy and difficult decisions were created by varying the similarity of S1 and S2. Statistical probability mapping showed enhanced gamma-band activity (GBA) over posterior parietal cortex for spatial and over left inferior frontal cortex for pattern changes (at approximately 120 to 220 ms after S2 onset). Activations were stronger for easy than difficult decisions. GBA over dorsolateral prefrontal cortex was more pronounced at approximately 280 to 430 ms for easy than difficult decisions regardless of type of change, possibly reflecting decision-relevant networks that integrate information from higher sensory areas representing the perceptual alternatives. Sensorimotor beta desynchronization as a measure of motor preparation peaked at approximately 460 ms for easy and at approximately 520 ms for difficult decisions, thus reflecting the reaction time difference between both conditions. In summary, the analysis of oscillatory activity in magnetoencephalogram served to elucidate the temporal dynamics of perceptual decision making in humans. PMID- 17138596 TI - Neural signatures of body ownership: a sensory network for bodily self consciousness. AB - Body ownership refers to the special perceptual status of one's own body, which makes bodily sensations seem unique to oneself. We studied the neural correlates of body ownership by controlling whether an external object was accepted as part of the body or not. In the rubber hand illusion (RHI), correlated visuotactile stimulation causes a fake hand to be perceived as part of one's own body. In the present study, we distinguished between the causes (i.e., multisensory stimulation) and the effect (i.e., the feeling of ownership) of the RHI. Participants watched a right or a left rubber hand being touched either synchronously or asynchronously with respect to their own unseen right hand. A quantifiable correlate of the RHI is a shift in the perceived position of the subject's hand toward the rubber hand. We used positron emission tomography to identify brain areas whose activity correlated with this proprioceptive measure of body ownership. Body ownership was related to activity in the right posterior insula and the right frontal operculum. Conversely, when the rubber hand was not attributed to the self, activity was observed in the contralateral parietal cortex, particularly the somatosensory cortex. These structures form a network that plays a fundamental role in linking current sensory stimuli to one's own body and thus also in self-consciousness. PMID- 17138597 TI - Brain mechanisms implicated in the preattentive categorization of speech sounds revealed using FMRI and a short-interval habituation trial paradigm. AB - A hallmark of categorical perception is better discrimination of stimulus tokens from 2 different categories compared with token pairs that are equally dissimilar but drawn from the same category. This effect is well studied in speech perception and represents an important characteristic of how the phonetic form of speech is processed. We investigated the brain mechanisms of categorical perception of stop consonants using functional magnetic resonance imaging and a passive short-interval habituation trial design (Zevin and McCandliss 2005). The paradigm takes advantage of neural adaptation effects to identify specific regions sensitive to an oddball stimulus presented in the context of a repeated item. These effects were compared for changes in stimulus characteristics that result in either a between-category (phonetic and acoustic) or a within-category (acoustic only) stimulus shift. Significantly greater activation for between category than within-category stimuli was observed in left superior sulcus and middle temporal gyrus as well as in inferior parietal cortex. In contrast, only a subcortical region specifically responded to within-category changes. The data suggest that these habituation effects are due to the unattended detection of a phonetic stimulus feature. PMID- 17138598 TI - Climate change--the greatest crisis for children? PMID- 17138599 TI - Oxymorphone hydrochloride, a potent opioid analgesic, is not carcinogenic in rats or mice. AB - Despite their long history of chronic use, little information is available regarding the carcinogenicity of opioid analgesics. Oxymorphone is a potent morphinan-type mu-opioid analgesic used for treatment of moderate-to-severe pain. Oxymorphone was tested for carcinogenicity in Crl:CD IGS BR rats and CD-1 mice. Oxymorphone hydrochloride was administered orally once daily for 2 years to rats at doses of 2.5, 5 and 10 mg/kg/day (males) and 5, 10 and 25 mg/kg/day (females), and mice at 10, 25, 75 and 150 mg/kg/day (65 animals per sex per group; 100 animals per sex in controls). In rats, survival was generally higher than controls in oxymorphone-treated groups, attributable to lower body weight gain. In mice, survival was generally higher than controls in females at all doses and males given < or = 25 mg/kg/day but lower in males given > or = 75 mg/kg/day due to a high incidence of obstructive uropathy. Opioid-related clinical signs and reduced body weight gain occurred in both species throughout the study. Nonneoplastic findings associated with oxymorphone pharmacology included ocular and pulmonary changes in rats considered secondary to inhibition of blinking and mydriasis, and antitussive activity, respectively, and urinary tract and renal findings in mice considered secondary to urinary retention. There was no target organ toxicity, and no increase in any neoplastic lesions attributed to oxymorphone. Plasma oxymorphone levels achieved in these studies exceeded those in patients taking high therapeutic doses of oxymorphone (Area under the curve [AUC(0-24 h)] values up to 5.6-fold and 64-fold in rats and mice, respectively). Oxymorphone is not considered to be carcinogenic in rats or mice under the conditions of these studies. PMID- 17138601 TI - Stability and plasticity of intrinsic membrane properties in hippocampal CA1 pyramidal neurons: effects of internal anions. AB - CA1 pyramidal neurons from animals that have acquired hippocampal tasks show increased neuronal excitability, as evidenced by a reduction in the postburst afterhyperpolarization (AHP). Studies of AHP plasticity require stable long-term recordings, which are affected by the intracellular solutions potassium methylsulphate (KMeth) or potassium gluconate (KGluc). Here we show immediate and gradual effects of these intracellular solutions on measurement of the AHP and basic membrane properties, and on the induction of AHP plasticity in CA1 pyramidal neurons from rat hippocampal slices. The AHP measured immediately after establishing whole-cell recordings was larger with KMeth than with KGluc. In general, the AHP in KMeth was comparable to the AHP measured in the perforated patch configuration. However, KMeth induced time-dependent changes in the intrinsic membrane properties of CA1 pyramidal neurons. Specifically, input resistance progressively increased by 70% after 50 min; correspondingly, the current required to trigger an action potential and the fast afterdepolarization following action potentials gradually decreased by about 50%. Conversely, these measures were stable in KGluc. We also demonstrate that activity-dependent plasticity of the AHP occurs with physiologically relevant stimuli in KGluc. AHPs triggered with theta-burst firing every 30 s were progressively reduced, whereas AHPs elicited every 150 s were stable. Blockade of the apamin-sensitive AHP current (I(AHP)) was insufficient to block AHP plasticity, suggesting that plasticity is manifested through changes in the apamin-insensitive slow AHP current (sI(AHP)). These changes were observed in the presence of synaptic blockers, and therefore reflect changes in the intrinsic properties of the neurons. However, no AHP plasticity was observed using KMeth. In summary, these data show that KMeth produces time-dependent changes in basic membrane properties and prevents or obscures activity-dependent reduction of the AHP. In whole-cell recordings using KGluc, repetitive theta-burst firing induced AHP plasticity that mimics learning-related reduction in the AHP. PMID- 17138602 TI - Electromechanical and pharmacomechanical signalling pathways for conducted vasodilatation along endothelium of hamster feed arteries. AB - Conducted vasodilatation (CVD) reflects the initiation and rapid (>mm s(-1)) spread of hyperpolarization along the endothelium and into smooth muscle. The ion channels that initiate CVD remain unclear as do signalling pathways that may complement electromechanical relaxation. Using isolated pressurized (75 mmHg; 37 degrees C) feed arteries (n=63; diameter: rest: 53 +/- 2 microm, maximal: 98 +/- 2 microm) from hamster retractor skeletal muscle, we investigated the contribution of calcium-activated potassium channels (KCa) and endothelium derived autacoids to CVD. Local delivery (1 microm micropipette tip; 500-2000 ms pulse) of acetylcholine (ACh) at the downstream end initiated a local increase in endothelial cell [Ca2+]i (Fura-PE3; Deltaratio 340/380 nm = 0.215 +/- 0.032) that preceded CVD along the entire vessel. During local perifusion with KCa antagonists, iberiotoxin (5 microm) had no effect, but charybdotoxin (CTX, 5 microm) + apamin (APA, 10 microm) abolished CVD reversibly. Remarkably, this local inhibition of KCa unmasked a 'slow-conducted vasodilatation' (SCVD) that spread >1200 microm at approximately 21 microm s(-1) (n=27). Recorded 500 microm upstream from the ACh stimulus, a rise in endothelial cell [Ca2+]i (Deltaratio 340/380 nm) = 0.146 +/- 0.017; P<0.05) preceded SCVD (Deltadiameter = 14 +/- 3 microm) by approximately 10 s. Before KCa inhibition, antagonism of nitric oxide synthase (Nomega-nitro-L-arginine, 250 microm; l-NNA) and cyclooxygenase (indomethacin, 5 microm; INDO) had no effect on the amplitude of CVD yet response duration decreased by one-third (P<0.05). During local CTX + APA perifusion, L NNA + INDO abolished SCVD while conducted [Ca2+]i responses remained intact. Thus, ACh triggers electromechanical relaxation of smooth muscle cells along the vessel initiated by local KCa, and the ensuing 'wave' of Ca2+ along the endothelium releases autacoids to promote pharmacomechanical relaxation. PMID- 17138603 TI - Ageing reduces nitric-oxide- and prostaglandin-mediated vasodilatation in exercising humans. AB - In older humans, infusions of endothelial agonists suggest endothelial dysfunction, due in part to less nitric oxide (NO)- and prostaglandin (PG) mediated vasodilatation, and a shift toward PG-mediated vasoconstriction. Ageing can also be associated with lower exercise blood flow (exercise hyperaemia), but the vascular mechanisms mediating this remain unknown. Notably, in young adults, inhibition of NO and PGs during exercise decreases exercise hyperaemia by approximately 20 and approximately 12%, respectively. We tested our first hypothesis that in older humans inhibition of NO would decrease hyperaemia, but that inhibition of PGs would increase hyperaemia by blocking vasoconstrictor PGs. Fifteen older subjects (65 +/- 3 years) performed dynamic forearm exercise for 20 min (20 contractions min(-1)). Forearm blood flow (FBF) was measured beat-to-beat with Doppler ultrasound, while saline or drugs were infused sequentially via brachial artery catheter in the exercising forearm. After achieving steady-state exercise, L-NAME (25 mg) was infused over 5 min to inhibit NO synthase. After a further 2 min of exercise (saline), ketorolac (6 mg) was infused over 5 min to inhibit PGs, followed by a further 3 min of exercise with saline. Drug order was reversed in seven subjects. L-NAME reduced steady-state exercise hyperaemia by 12 +/- 3% in older subjects (P<0.01), whereas ketorolac had no net effect on blood flow (3 +/- 6%, P>0.4). The effects of l-NAME and ketorolac were independent of drug order. By comparing these results with our previous results in young adults, we tested our second hypothesis that in older humans inhibition of NO or PGs would have less impact on exercise hyperaemia due to less vasodilatation from these signals. Our results suggest that, compared with young adults, in older humans the relative contribution of NO to exercise hyperaemia is reduced approximately 45% (22 +/- 4 versus 12 +/- 3%), but the role of PG in mediating vasodilatation is lost in ageing human skeletal muscle. Lower exercise hyperaemia in older humans may be mediated in part by less NO- and PG-mediated vasodilatation during exercise. PMID- 17138604 TI - Window Ca2+ current and its modulation by Ca2+ release in hypertrophied cardiac myocytes from dogs with chronic atrioventricular block. AB - Torsades de pointes (TdP) ventricular tachycardia typically occurs in the setting of early afterdepolarizations; it contributes to arrhythmias and sudden death in congenital and acquired heart disease. Window L-type Ca2+ current (ICaL) has a central role in the arrhythmogenesis and may be particularly important under beta adrenergic stimulation. We studied the properties of ICaL in myocytes from the dog with chronic atrioventricular block (cAVB) that has cardiac hypertrophy and an increased susceptibility to TdP. Peak ICaL densities at baseline (K+ - and Na+ -free solutions, 10 mmol l(-1) [EGTA]pip) in cAVB were comparable to control, but inactivation was shifted to the right, resulting in a larger window current area in cAVB. Under beta-adrenergic stimulation, the window current area was increased and shifted to the left, but less so in cAVB (maximum at -27 mV, versus -32 mV in control). ICaL during a step to -35 mV showed a transient reduction immediately after the peak. Test steps to 0 mV, simultaneous recording of [Ca2+]i and manipulation of sarcoplasmic reticulum (SR) Ca2+ release showed that this resulted from inhibition and fast recovery of ICaL with SR Ca2+ release. The extent of this dynamic modulation was larger in cAVB than in control (23 +/- 2% of the initially available current, versus 13 +/- 3%; P<0.05). Early afterdepolarizations (EADs) in cAVB myocytes under beta-adrenergic stimulation typically occurred in the window current voltage range and after decline of [Ca2+]i. In conclusion, in cAVB, the larger window current, its rightward shift and enhanced dynamic modulation by SR Ca2+ release may contribute to an increased incidence of EADs in cAVB under beta-adrenergic stimulation. PMID- 17138605 TI - Acceleration of AMPA receptor kinetics underlies temperature-dependent changes in synaptic strength at the rat calyx of Held. AB - It is well established that synaptic transmission declines at temperatures below physiological, but many in vitro studies are conducted at lower temperatures. Recent evidence suggests that temperature-dependent changes in presynaptic mechanisms remain in overall equilibrium and have little effect on transmitter release at low transmission frequencies. Our objective was to examine the postsynaptic effects of temperature. Whole-cell patch-clamp recordings from principal neurons in the medial nucleus of the trapezoid body showed that a rise from 25 degrees C to 35 degrees C increased miniature EPSC (mEPSC) amplitude from -33 +/- 2.3 to -46 +/- 5.7 pA (n=6) and accelerated mEPSC kinetics. Evoked EPSC amplitude increased from -3.14 +/- 0.59 to -4.15 +/- 0.73 nA with the fast decay time constant accelerating from 0.75 +/- 0.09 ms at 25 degrees C to 0.56 +/- 0.08 ms at 35 degrees C. Direct application of glutamate produced currents which similarly increased in amplitude from -0.76 +/- 0.10 nA at 25 degrees C to -1.11 +/- 0.19 nA 35 degrees C. Kinetic modelling of fast AMPA receptors showed that a temperature-dependent scaling of all reaction rate constants by a single multiplicative factor (Q10=2.4) drives AMPA channels with multiple subconductances into the higher-conducting states at higher temperature. Furthermore, Monte Carlo simulation and deconvolution analysis of transmission at the calyx of Held showed that this acceleration of the receptor kinetics explained the temperature dependence of both the mEPSC and evoked EPSC. We propose that acceleration in postsynaptic AMPA receptor kinetics, rather than altered presynaptic release, is the primary mechanism by which temperature changes alter synaptic responses at low frequencies. PMID- 17138606 TI - Increased responsiveness of rat colonic splanchnic afferents to 5-HT after inflammation and recovery. AB - 5-Hydroxytryptamine (5-HT) activates colonic splanchnic afferents, a mechanism by which it has been implicated in generating symptoms in postinfectious and postinflammatory states in humans. Here we compared mechanisms of colonic afferent activation by 5-HT and mechanical stimuli in normal and inflamed rat colon, and after recovery from inflammation. Colonic inflammation was induced in rats by dextran sulphate sodium. Single-fibre recordings of colonic lumbar splanchnic afferents revealed that 58% of endings responded to 5-HT (10(-4) m) in controls, 88% in acute inflammation (P<0.05) and 75% after 21 days recovery (P < 0.05 versus control). Maximal responses to 5-HT were also larger, and the estimated EC50 was reduced from 3.2 x 10(-6) to 8 x 10(-7) m in acute inflammation and recovered to 2 x 10(-6) m after recovery. Responsiveness to mechanical stimulation was unaffected. 5-HT3 receptor antagonism with alosetron reduced responses to 5-HT in controls but not during inflammation. Responses to the mast cell degranulator 48/80 mimicked those to 5-HT in inflamed tissue but not in controls, and more 5-HT-containing mast cells were seen close to calcitonin gene-related peptide-containing fibres in inflamed serosa. We conclude that colonic serosal and mesenteric endings exhibit increased sensitivity to 5-HT in inflammation, with both an increase in proportion of responders and an increase in sensitivity, which is maintained after healing of inflammation. This is associated with alterations in the roles of 5-HT3 receptors and mast cells. PMID- 17138607 TI - Removal of phosphorylation sites of gamma subunit of phosphodiesterase 6 alters rod light response. AB - The phosphodiesterase 6 gamma (PDE6 gamma) inhibitory subunit of the rod PDE6 effector enzyme plays a central role in the turning on and off of the visual transduction cascade, since binding of PDE6 gamma to the transducin alpha subunit (T alpha) initiates the hydrolysis of the second messenger cGMP, and PDE6 gamma in association with RGS9-1 and the other GAP complex proteins (G beta 5, R9AP) accelerates the conversion of T alpha GTP to T alpha GDP, the rate-limiting step in the decay of the rod light response. Several studies have shown that PDE6 gamma can be phosphorylated at two threonines, T22 and T35, and have proposed that phosphorylation plays some role in the physiology of the rod. We have examined this possibility by constructing mice in which T22 and/or T35 were replaced with alanines. Our results show that T35A rod responses rise and decay more slowly and are less sensitive to light than wild-type (WT). T22A responses show no significant difference in initial time course with WT but decay more rapidly, especially at dimmer intensities. When the T22A mutation is added to T35A, double mutant rods no longer showed the prolonged decay of T35A rods but remained slower than WT in initial time course. Our experiments suggest that the polycationic domain of PDE6 gamma containing these two phosphorylation sites can influence the rate of PDE6 activation and deactivation and raise the possibility that phosphorylation or dephosphorylation of PDE6 gamma could modify the time course of transduction, thereby influencing the wave form of the light response. PMID- 17138608 TI - The electrogenic Na+/HCO3- cotransport modulates resting membrane potential and action potential duration in cat ventricular myocytes. AB - Perforated whole-cell configuration of patch clamp was used to determine the contribution of the electrogenic Na+/HCO3- cotransport (NBC) on the shape of the action potential in cat ventricular myocytes. Switching from Hepes to HCO3- buffer at constant extracellular pH (pH(o)) hyperpolarized resting membrane potential (RMP) by 2.67 +/- 0.42 mV (n = 9, P < 0.05). The duration of action potential measured at 50% of repolarization time (APD50) was 35.8 +/- 6.8% shorter in the presence of HCO3- than in its absence (n = 9, P < 0.05). The anion blocker SITS prevented and reversed the HCO3- -induced hyperpolarization and shortening of APD. In addition, no HCO3- -induced hyperpolarization and APD shortening was observed in the absence of extracellular Na+. Quasi-steady-state currents were evoked by 8 s duration voltage-clamped ramps ranging from -130 to +30 mV. A novel component of SITS-sensitive current was observed in the presence of HCO3-. The HCO3- -sensitive current reversed at -87 +/- 5 mV (n = 7), a value close to the expected reversal potential of an electrogenic Na+/HCO3- cotransport with a HCO3-:Na+ stoichiometry ratio of 2: 1. The above results allow us to conclude that the cardiac electrogenic Na+/HCO3- cotransport has a relevant influence on RMP and APD of cat ventricular cells. PMID- 17138609 TI - Role of myosin heavy chain composition in the stretch activation response of rat myocardium. AB - The speed and force of myocardial contraction during systolic ejection is largely dependent on the intrinsic contractile properties of cardiac myocytes. As the myosin heavy chain (MHC) isoform of cardiac muscle is an important determinant of the contractile properties of individual myocytes, we studied the effects of altered MHC isoform expression in rat myocardium on the mechanical properties of skinned ventricular preparations. Skinned myocardium from thyroidectomized rats expressing only the beta MHC isoform displayed rates of force redevelopment that were about 2.5-fold slower than in myocardium from hyperthyroid rats expressing only the alpha MHC isoform, but the amount of force generated at a given level of Ca2+ activation was not different. Because recent studies suggest that the stretch activation response in myocardium has an important role in systolic function, we also examined the effect of MHC isoform expression on the stretch activation response by applying a rapid stretch (1% of muscle length) to an otherwise isometrically contracting muscle fibre. Sudden stretch of myocardium resulted in a concomitant increase in force that quickly decayed to a minimum and was followed by a delayed redevelopment of force (i.e. stretch activation) to levels greater than prestretch force. beta MHC expression dramatically slowed the overall rate of the stretch activation response compared to expression of alpha MHC isoform; specifically, the rate of force decay was approximately 2-fold slower and the rate of delayed force development was approximately 2.5-fold slower. In contrast, MHC isoform had no effect on the amplitude of the stretch activation response. Collectively, these data show that expression of beta MHC in myocardium dramatically slows rates of cross-bridge recruitment and detachment which would be expected to decrease power output and contribute to depressed systolic function in end-stage heart failure. PMID- 17138610 TI - TRPpathies. AB - Many human diseases are caused by mutations in ion channels. Dissecting the pathogenesis of these 'channelopathies' has yielded important insights into the regulation of vital biological processes by ions and has become a productive tool of modern ion channel biology. One of the best examples of a synergism between the clinical and basic science aspects of a modern biological topic is cystic fibrosis. Not only did the identification of the ion channel mutated in cystic fibrosis pinpoint the root cause of this disease, but it also has significantly advanced our understanding of basic biological processes as diverse as protein folding and epithelial fluid and electrolyte secretion. The list of confirmed 'channelopathies' is growing and several members of the TRP family of ion channels have been implicated in human diseases such as mucolipidosis type IV (MLIV), autosomal dominant polycystic kidney disease (ADPKD), familial focal segmental glomerulosclerosis (FSG), hypomagnesemia with secondary hypocalcaemia (HSH), and several forms of cancer. Analysing pathogenesis of the diseases linked to TRP dysregulation provides an exciting means of identifying novel functions of TRP channels. PMID- 17138611 TI - Lack of involvement of the autonomic nervous system in early ventilatory and pulmonary vascular acclimatization to hypoxia in humans. AB - The activity within the autonomic nervous system may be altered following sustained exposure to hypoxia, and it is possible that this increase in activity underlies the early acclimatization of both ventilation and the pulmonary vasculature to hypoxia. To test this hypothesis, seven individuals were infused with the ganglionic blocker trimetaphan before and after an 8 h exposure to hypoxia. The short half-life of trimetaphan should ensure that the initial infusion does not affect acclimatization to the 8 h hypoxia exposure, and the use of a ganglion blocking agent should inhibit activity within all branches of the autonomic nervous system. During the infusions of trimetaphan, measurements of ventilation and echocardiographic assessments of pulmonary vascular tone (DeltaPmax) were made during euoxia and during a short period of isocapnic hypoxia. Subjects were also studied on two control days, when a saline infusion was substituted for trimetaphan. Trimetaphan had no effect on either euoxic ventilation or the sensitivity of ventilation to acute hypoxia. Trimetaphan significantly reduced DeltaPmax in euoxia (P<0.05), but had no significant effect on the sensitivity of DeltaPmax to acute hypoxia once changes in cardiac output had been controlled for. The 8 h period of hypoxia elevated euoxic ventilation (P<0.001) and DeltaPmax (P<0.001) and increased their sensitivities to acute hypoxia (P<0.001 for both), indicating that significant acclimatization had occurred. Trimetaphan had no effect on the acclimatization response of any of these variables. We conclude that altered autonomic activity following 8 h of hypoxia does not underlie the acclimatization observed in ventilation or pulmonary vascular tone. PMID- 17138612 TI - Beliefs, recommendations and intentions are important explanatory factors of mammography screening behavior among Muslim Arab women in Israel. AB - The rates of mammography screening by Muslim Arab women in Israel are lower compared with the general population. The current study aimed to examine factors related to screening mammography behavior among Arab women by employing components from the Health Belief Model and the Theory of Reasoned Action. Sociodemographic factors, knowledge, beliefs about breast cancer and mammography, self-efficacy, cues to action, norms and intention to perform mammography were examined as explanatory variables for mammography use. Face-to-face interviews with a random sample of 510 Muslim Arab women, aged 50-69 years, were conducted. The women had limited knowledge about breast cancer and mammography, and the rate of mammography screening behavior (at the recommended interval) was only 20%. The women who were significantly more likely to undergo mammography were those who received a recommendation from a health professional or from family/friends, perceived themselves as vulnerable to getting breast cancer, believed in the efficacy of the test, perceived it as not painful, were younger, were more educated and were only of borderline significance among those who expressed an intention to undergo mammography. The findings indicate that professional recommendation and beliefs sets are essential factors for developing effective mammography screening interventions in this unique population. PMID- 17138613 TI - Validating the theoretical structure of the Treatment Self-Regulation Questionnaire (TSRQ) across three different health behaviors. AB - Nearly 40% of mortality in the United States is linked to social and behavioral factors such as smoking, diet and sedentary lifestyle. Autonomous self-regulation of health-related behaviors is thus an important aspect of human behavior to assess. In 1997, the Behavior Change Consortium (BCC) was formed. Within the BCC, seven health behaviors, 18 theoretical models, five intervention settings and 26 mediating variables were studied across diverse populations. One of the measures included across settings and health behaviors was the Treatment Self-Regulation Questionnaire (TSRQ). The purpose of the present study was to examine the validity of the TSRQ across settings and health behaviors (tobacco, diet and exercise). The TSRQ is composed of subscales assessing different forms of motivation: amotivation, external, introjection, identification and integration. Data were obtained from four different sites and a total of 2731 participants completed the TSRQ. Invariance analyses support the validity of the TSRQ across all four sites and all three health behaviors. Overall, the internal consistency of each subscale was acceptable (most alpha values >0.73). The present study provides further evidence of the validity of the TSRQ and its usefulness as an assessment tool across various settings and for different health behaviors. PMID- 17138614 TI - Understanding physical activity participation in members of an African American church: a qualitative study. AB - Faith-based interventions hold promise for increasing physical activity (PA) and thereby reducing health disparities. This paper examines the perceived influences on PA participation, the link between spirituality and health behaviors and the role of the church in promoting PA in African Americans. Participants (n = 44) were adult members of African American churches in South Carolina. In preparation for a faith-based intervention, eight focus groups were conducted with sedentary or underactive participants. Groups were stratified by age (<55 years versus >or=55 years), geography and gender. Four general categories were determined from the focus groups: spirituality, barriers, enablers and desired PA programs. Personal, social, community and environmental barriers and enablers were described by both men and women, with no apparent differences by age. Additionally, both men and women mentioned aerobics, walking programs, sports and classes specifically for older adults as PA programs they would like available at church. This study provides useful information for understanding the attitudes and experiences with exercise among African Americans, and provides a foundation for promoting PA through interventions with this population by incorporating spirituality, culturally specific activities and social support within the church. PMID- 17138615 TI - Factors influencing the dietary response to a nutritional intervention promoting the Mediterranean food pattern in healthy women from the Quebec City metropolitan area. AB - The purpose of this study was to evaluate the influence of sociodemographic characteristics and baseline food habits on the dietary response to a nutritional intervention promoting the Mediterranean food pattern and maintenance of dietary modifications in 73 healthy women. The 12-week nutritional intervention in free living conditions consisted of two group courses and seven individual sessions with a dietitian. A follow-up visit was performed 12 weeks after the end of the intervention (week 24). A Mediterranean dietary score was derived from a food frequency questionnaire, administered at 0, 6, 12 and 24 weeks. Marital status, socioeconomic level, educational level and household size did not seem to influence the dietary response, whereas women without children followed more closely dietary advice than women with children (OR, 3.6; 95% CI, 1.3-10.0). Planning food purchases in function of weekly discounts was also associated with better dietary response to the intervention (OR, 3.3; 95% CI, 1.3-8.8). Nutritional intervention promoting the Mediterranean food pattern was effective in modifying food habits of healthy women. The fact of having children or not and food purchase habits seem to influence the response to a nutritional intervention promoting the Mediterranean food pattern. PMID- 17138616 TI - The Behavior-Image Model: a paradigm for integrating prevention and health promotion in brief interventions. AB - This paper describes the Behavior-Image Model (BIM), an emerging and innovative paradigm for planning brief interventions for adolescents that fuse the prevention of harmful behaviors with the promotion of healthy habits. We discuss the components of the BIM as a new paradigm for creating multiple behavior health interventions, as well as the empirical and conceptual underpinnings of the model, and present Project Sport as an illustration of how the BIM may be applied to construct a brief multi-behavior intervention. The BIM posits that selected salient images of others and ourselves may be used to cast gain- and loss-framed messages coupling and motivating health-promoting and health-risk behaviors within single interventions. This content in turn activates prototypes and future self-images through the processes of social and self-comparison, leading to improvements in risk and protective factors and subsequent change in targeted health-promoting and health-risk behaviors. Recommendations are offered for conducting future research integrating health-risk and health-promoting behaviors in both brief and non-brief interventions for adolescents and adults. PMID- 17138617 TI - Disease-specific health education for COPD: a systematic review of changes in health outcomes. AB - A systematic review was conducted to determine the benefits of disease-specific health education for people with chronic obstructive pulmonary disease (COPD). A search was conducted through Medline, CINAHL, PsycINFO, Embase, Cochrane Library, Physiotherapy Evidence Database and reference lists to obtain publications reporting on educational interventions compared with usual medical care. Two reviewers independently assessed each paper for methodological quality and data extraction. Thirteen publications describing 10 randomized controlled trials were identified for inclusion. The studies reported on a very broad variety of outcomes and follow-up periods, making a meta-analysis not possible for most measures. Didactical educational intervention for the COPD population appeared to have minimal effect on health outcomes including quality of life, health care utilization, exercise capacity or lung function and is therefore not the education delivery method recommended. Education focusing on self-management showed encouraging results with a tendency for improvements in quality of life and health care utilization, but the results did not reach statistical significance as sample sizes were insufficient to detect an effect. Further research is required into self-management education as it is not possible to make generalizations from the current published literature as to the benefits in changing health status in the COPD population. PMID- 17138618 TI - Ultrastructural changes and sarcoplasmic reticulum Ca2+ regulation in red vastus muscle following eccentric exercise in the rat. AB - This study examined the effects of a bout of low-intensity, prolonged downhill exercise on sarcoplasmic reticulum (SR) Ca(2+)-ATPase activity, Ca(2+) uptake and release in rat red vastus muscle. Ionophore stimulation was determined to assess vesicle integrity by measuring the ratio of Ca(2+)-ATPase activities in the presence and absence of A23187. Observations of the muscle ultrastructure were made to evaluate muscle damage at the level of the myofibrils and SR. Adult male Sprague-Dawley rats (weight, 395 +/- 5.9 g) were either assigned as non-exercise controls or subjected to 90 min of downhill treadmill exercise (-16 deg; 15 m min(-1)), and then killed immediately, 4, 24, 48, 72 or 144 h after exercise (n = 7). Calcium uptake was significantly lower (P < 0.05) compared with control values (19.25 +/- 1.38 nmol min(-1) (mg protein)(-1)), by 29 and 36% immediately and 4 h postexercise, respectively, and remained depressed (P < 0.05) 24 h postexercise. Calcium release was also significantly lower (P < 0.05) compared with control values (31.06 +/- 2.36 nmol min(-1) (mg protein)(-1)), by 37 and 39% immediately and 4 h postexercise, respectively, and remained depressed (P < 0.05) 24 h postexercise. Ca(2+)-ATPase activity measured with ionophore was 31% lower (P < 0.05) 4 h postexercise, and remained lower (P < 0.05) 24 h postexercise. The ratio of Ca(2+)-ATPase activities in the presence and absence of A23187 was not significantly changed after exercise, indicating that membrane integrity was not altered by the exercise. Focal dilatations of the SR were observed immediately and 4 h following exercise, implying that SR may be susceptible to damage in the localized regions of overstretched sarcomeres. The results demonstrate that a bout of low-intensity, prolonged downhill exercise results in a long-lasting depression of SR function that is not fully restored after 2 days of recovery, which may underlie some functional impairments induced by eccentric exercise. PMID- 17138619 TI - Modelling the glucocorticoid receptor and producing therapeutic agents with anti inflammatory effects but reduced side-effects. AB - Glucocorticoid hormones exert a wide spectrum of metabolic and immunological effects. They are synthesized from a cholesterol precursor and are structurally related to the other steroid hormones, progesterone, aldosterone and oestrogen. They act through the glucocorticoid receptor (GR), a member of the nuclear receptor superfamily. The GR is an intracellular receptor; the hydrophobic ligand accesses its receptor by diffusion across the plasma membrane. The ligand activated GR translocates to the nucleus to regulate expression of its target genes. The GR, in common with the rest of the receptor family, can be functionally divided into an N-terminal transcription activation domain, a central DNA binding domain and a C-terminal ligand binding domain, which also includes a second transactivation domain. Although synthetic glucocorticoids are the most potent anti-inflammatory agents known, their use is limited owing to the range and severity of their side-effects. The structure of the ligand binding domain of the glucocorticoid receptor has now been solved, and a series of studies has shown that even subtle changes to the ligand structure alter the final conformation of the ligand-receptor complex, with consequences for further protein recruitment and for the function of the receptor. This, coupled with the successful development of selective oestrogen receptor agonists, has led to concerted efforts to find selective GR ligands, with preserved beneficial anti inflammatory activity, but reduced side-effect profile. Current efforts have identified several useful tool compounds, and further molecules are in development in several pharmaceutical companies. PMID- 17138620 TI - Differential sensitivity of excitatory and inhibitory synaptic transmission to modulation by nitric oxide in rat nucleus tractus solitarii. AB - The nucleus tractus solitarii (NTS) is a key central link in control of multiple homeostatic reflexes. A number of studies have demonstrated that exogenous and endogenous nitric oxide (NO) within NTS regulates visceral function, but further understanding of the role of NO in the NTS is hampered by the lack of information about its intracellular actions. We studied effects of NO in acute rat brainstem slices. Aqueous NO solution (NO(aq)) potentiated electrically evoked excitatory and inhibitory postsynaptic potentials (EPSPs and IPSPs, respectively) in different neuronal subpopulations and, in some neurones, caused a depolarization. Similar effects were observed using the NO donor diethylamine NONOate (DEA/NO). The threshold NO concentration as determined using an NO electrochemical sensor was estimated as approximately 0.4 nm (EC(50) approximately 0.9 nm) for potentiating glutamatergic EPSPs but approximately 3 nm for monosynaptic GABAergic IPSPs. Bath application of the soluble guanylate cyclase (sGC) inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) abolished NO(aq)- and DEA/NO-induced potentiation of evoked EPSPs, IPSPs and depolarization. All NO actions were mimicked by the non-NO-dependent guanylate cyclase activator Bay 41 2272. The effects of NO on EPSPs and IPSPs persisted in cells where postsynaptic sGC was blocked by ODQ and therefore were presynaptic, owing to a direct modulation of transmitter release combined with depolarization of presynaptic neurones. Therefore, while lower concentrations of NO may be important for fine tuning of glutamatergic transmission, higher concentrations are required to directly engage GABAergic inhibition. This differential sensitivity of excitatory and inhibitory connections to NO may be important for determining the specificity of the effects of this freely diffusible gaseous messenger. PMID- 17138621 TI - High-frequency oscillations in phrenic activity during pontile and medullary respiratory rhythms in rats. AB - High-frequency oscillations may be signatures of the basic mechanisms underlying the neurogenesis of various patterns of automatic ventilatory activity. These high-frequency oscillations in phrenic activity differ greatly in eupnoea and gasping, implying different mechanisms of neurogenesis. In a decerebrate, in situ preparation of the rat, the peak frequency of high-frequency oscillations fell in apneusis following removal of the rostral pons. Following removal of all pons, phrenic discharge had a mixed pattern of gasps and multiple bursts; some of the latter were incrementing, as in eupnoea. Regardless of pattern, peak frequencies were significantly below those which were found during eupnoea, apneusis or gasping of the decerebrate preparation. Results do not support the concept that 'non-gasping' rhythmic patterns that can be recorded following a removal of pons are generated by the same mechanisms as those generating eupnoea. Indeed, both pons and medulla appear essential for all aspects of eupnoea to be expressed. PMID- 17138622 TI - Acute intermittent hypoxia increases both phrenic and sympathetic nerve activities in the rat. AB - The respiratory system expresses multiple forms of plasticity, defined as alterations in the breathing pattern that persist or develop after a stimulus. Stimulation of breathing with intermittent hypoxia (IH) elicits long-term facilitation (LTF), a type of plasticity in which respiratory motor activity progressively increases in anaesthetized animals, even after the stimuli have ceased and blood gases have normalized. It is unknown whether the sympathetic nervous system similarly expresses IH-induced plasticity, but we predicted that IH would evoke LTF in sympathetic nerve activity (SNA) because respiratory and sympathetic control systems are coupled. To test this idea, we recorded splanchnic (sSNA) and phrenic nerve activities (PNA) in equithesin-anaesthetized rats. Animals were exposed to 10 45 s episodes of 8% O(2)-92% N(2), separated by 5 min intervals of 100% O(2), and recordings were continued for 60 min following the last hypoxic exposure. Cycle-triggered averages of integrated PNA and sSNA from periods preceding, and 5 and 60 min following the hypoxic stimuli were compared. Intermittent hypoxia significantly increased both sSNA and PNA. Treatment with methysergide (3 mg kg(-1), i.v.) 20 min before the intermittent hypoxic exposures prevented the increases in integrated PNA and sSNA 60 min after IH, indicating a role of serotonergic pathways in this form of plasticity. No increases in PNA and sSNA occurred at comparable times (60 and 120 min) in rats not exposed to hypoxia. The increased sSNA was not simply tonic, but was correlated with respiratory bursts, and occurred predominantly during the first half of expiration. These findings support the hypothesis that sympathorespiratory coupling may underlie the sustained increase in SNA associated with the IH that occurs during sleep apnoea. PMID- 17138623 TI - Autonomic regulation during orthostatic stress in highlanders: comparison with sea-level residents. AB - This report is a comparison of orthostatic tolerance and autonomic function in three groups of high-altitude dwellers: Andeans with and without chronic mountain sickness (CMS) and healthy Ethiopians. Results are compared with those from healthy sea-level residents. The aim was to determine whether different high altitude populations adapted differently to the prevailing hypobaric hypoxia. Orthostatic tolerance was assessed using a test involving head-up tilt (HUT) and graded lower body suction. This was performed at the subjects' resident altitude. Blood pressure (Portapres) and R-R interval (ECG) were recorded during the test, and spectral and cross-spectral analyses of heart period and systolic blood pressure time series were performed using data obtained both while supine and during HUT. The transfer function gain in the low-frequency range (LF, approximately 0.1 Hz) at the point of maximal coherence was used as a measure of cardiac baroreflex sensitivity (BRS). As previously reported, Peruvians displayed an unusually good orthostatic tolerance, while Ethiopians showed an orthostatic tolerance comparable to that of healthy sea-level residents. There were no significant differences between groups in the supine values of the spectral analysis results. Head-up tilt induced the expected changes in Ethiopians (an increase in the LF components and a decrease in the respiratory components) but not in Andeans. Cross-spectral analysis showed abnormal results from all groups of high-altitude dwellers. These results indicate that Ethiopians, but not Peruvians, behave similarly to sea-level residents in terms of orthostatic tolerance and autonomic responses to orthostatic stress, as assessed from spectral analyses, and this indicates good adaptation to their environment. However, in all the high-altitude groups the results of cross-spectral analysis were atypical, suggesting some degree of impairment in baroreflex function. PMID- 17138624 TI - Visualization of dynamics of plant-pathogen interaction by novel combination of chlorophyll fluorescence imaging and statistical analysis: differential effects of virulent and avirulent strains of P. syringae and of oxylipins on A. thaliana. AB - Pathogen infection leads to defence induction as well as to changes in carbohydrate metabolism of plants. Salicylic acid and oxylipins are involved in the induction of defence, but it is not known if these signalling molecules also mediate changes in carbohydrate metabolism. In this study, the effect of application of salicylic acid and the oxylipins 12-oxo-phytodienoic acid (OPDA) and jasmonic acid on photosynthesis was investigated by kinetic chlorophyll fluorescence imaging and compared with the effects of infection by virulent and avirulent strains of Pseudomonas syringae. Both pathogen strains and OPDA caused a similar change in fluorescence parameters of leaves of Arabidopsis thaliana. The response to OPDA appeared faster compared with that to the pathogens and persisted only for a short time. Infiltration with jasmonic acid or salicylic acid did not lead to a localized and distinct fluorescence response of the plant. To capture the faint early symptoms of the plant response, a novel algorithm was applied identifying the unique fluorescence signature-the set of images that, when combined, yield the highest contrast between control and infected leaf segments. Unlike conventional fluorescence parameters, this non-biased approach indeed detected the infection as early as 6 h after inoculation with bacteria. It was posssible to identify distinct fluorescence signatures characterizing the early and late phases of the infection. Fluorescence signatures of both infection phases were found in leaves infiltrated with OPDA. PMID- 17138625 TI - The role of the sucrose transporter, OsSUT1, in germination and early seedling growth and development of rice plants. AB - Using expression analysis, the role of the sucrose transporter OsSUT1 during germination and early growth of rice seedlings has been examined in detail, over a time-course ranging from 1 d to 7 d post-imbibition. Unlike the wheat orthologue, TaSUT1, which is thought to be directly involved in sugar transfer across the scutellar epithelium, OsSUT1 is not expressed in the scutellar epithelial cell layer of germinating rice and is, therefore, not involved in transport of sugars across the symplastic discontinuity between the endosperm and the embryo. OsSUT1 expression was also absent from the aleurone cells, indicating it is not involved in the transport of sucrose in this cell layer during germination. However, by 3 d post-imbibition, OsSUT1 was present in the companion cells and sieve elements of the scutellar vascular bundle, where it may play a role in phloem loading of sucrose for transport to the developing shoot and roots. This sucrose is most likely sourced from hexoses imported from the endosperm. In addition, sucrose may be remobilized from starch granules which are present at a high density in the scutellar ground tissues surrounding the vasculature and at the base of the shoot. OsSUT1 was also present in the coleoptile and the first and second leaf blades, where it was localized to the phloem along the entire length of these tissues, and was also present within the phloem of the primary roots. OsSUT1 may be involved in retrieval of sugars from the apoplasm in these tissues. PMID- 17138626 TI - A new evolutionary paradigm for the Parkinson disease gene DJ-1. AB - The DJ-1 gene is extensively studied because of its involvement in familial Parkinson disease. DJ-1 belongs to a complex superfamily of genes that includes both prokaryotic and eukaryotic representatives. We determine that many prokaryotic groups, such as proteobacteria, cyanobacteria, spirochaetes, firmicutes, or fusobacteria, have genes, often incorrectly called "Thij," that are very close relatives of DJ-1, to the point that they cannot be clearly separated from the eukaryotic DJ-1 genes by phylogenetic analyses of their sequences. In addition, and contrary to a previous study that suggested that DJ-1 genes were animal specific, we show that DJ-1 genes are found in at least 5 of the 6 main eukaryotic groups: opisthokonta (both animals and fungi), plantae, chromalveolata, excavata, and amoebozoa. Our results thus provide strong evidence for DJ-1 genes originating before the origin of eukaryotes. Interestingly, we found that some fungal species, among them the model yeast Schizosaccharomyces pombe, have DJ-1-like genes, most likely orthologous to the animal genes. This finding opens new ways for the analysis of the functions of this group of genes. PMID- 17138627 TI - Increasing sequence correlation limits the efficiency of recombination in a multisite evolution model. AB - The accumulation of preexisting beneficial alleles in a haploid population, under selection and infrequent recombination, and in the absence of new mutation events is studied numerically by means of detailed Monte Carlo simulations. On the one hand, we confirm our previous work, in that the accumulation rate follows modified single-site kinetics, with a timescale set by an effective selection coefficient s(eff) as shown in a previous work, and we confirm the qualitative features of the dependence of s(eff) on the population size and the recombination rate reported therein. In particular, we confirm the existence of a threshold population size below which evolution stops before the emergence of best-fit individuals. On the other hand, our simulations reveal that the population dynamics is essentially shaped by the steady accumulation of pairwise sequence correlation, causing sequence congruence in excess of what one would expect from a uniformly random distribution of alleles. By sequence congruence, we understand here the opposite of genetic distance, that is, the fraction of monomorphic sites of specified allele type in a pair of genomes (individual sequences). The effective selection coefficient changes more rapidly with the recombination rate and has a higher threshold in this parameter than found in the previous work, which neglected correlation effects. We examine this phenomenon by monitoring the time dependence of sequence correlation based on a set of sequence congruence measures and verify that it is not associated with the development of linkage disequilibrium. We also discuss applications to HIV evolution in infected individuals and potential implications for drug therapy. PMID- 17138628 TI - Distal tears of the hamstring muscles: review of the literature and our results of surgical treatment. AB - BACKGROUND: Hamstring strains are among the most frequent injuries in sports, especially in events requiring sprinting and running. Distal tears of the hamstring muscles requiring surgical treatment are scarcely reported in the literature. OBJECTIVE: To evaluate the results of surgical treatment for distal hamstring tears. DESIGN: A case series of 18 operatively treated distal hamstring muscle tears combined with a review of previously published cases in the English literature. Retrospective study; level of evidence 4. SETTING: Mehilainen Sports Trauma Research Center, Mehilainen Hospital and Sports Clinic, Turku, Finland. PATIENTS: Between 1992 and 2005, a total of 18 athletes with a distal hamstring tear were operated at our centre. MAIN OUTCOME MEASUREMENTS: At follow-up, the patients were asked about possible symptoms (pain, weakness, stiffness) and their return to the pre-injury level of sport. RESULTS: The final results were rated excellent in 13 cases, good in 1 case, fair in 3 cases and poor in 1 case. 14 of the 18 patients were able to return to their former level of sport after an average of 4 months (range 2-6 months). CONCLUSIONS: Surgical treatment seems to be beneficial in distal hamstring tears in selected cases. PMID- 17138629 TI - Injuries in amateur horse racing (point to point racing) in Great Britain and Ireland during 1993-2006. AB - OBJECTIVES: To provide a breakdown of injury incidence from amateur jump racing (also known as point to point racing) in Great Britain and Ireland during 1993 2006 and to compare the injury epidemiology with professional horse racing in Great Britain, Ireland and France. DESIGN: Retrospective review. SETTING: Great Britain and Ireland. PARTICIPANTS: Amateur jockeys. MAIN OUTCOME MEASURES: Injury rates. RESULTS: Injury data suggest that point to point racing is more dangerous from an injury point of view than professional jump racing, which has previously been shown to be more dangerous than flat racing. Amateur jockeys have more falls than their professional counterparts, and this in turn puts them at greater risk of sustaining more serious injuries. CONCLUSIONS: Amateur (point to point) jockeys represent a sporting population that previously has been little studied. They represent a group at high risk of injury, and hence formal injury surveillance tracking and counter measures for injury prevention are recommended. PMID- 17138630 TI - Time-motion analysis and physiological data of elite under-19-year-old basketball players during competition. AB - The physical demands of modern basketball were assessed by investigating 38 elite under-19-year-old basketball players during competition. Computerised time-motion analyses were performed on 18 players of various positions. Heart rate was recorded continuously for all subjects. Blood was sampled before the start of each match, at half time and at full time to determine lactate concentration. Players spent 8.8% (1%), 5.3% (0.8%) and 2.1% (0.3%) of live time in high "specific movements", sprinting and jumping, respectively. Centres spent significantly lower live time competing in high-intensity activities than guards (14.7% (1%) v 17.1% (1.2%); p<0.01) and forwards (16.6% (0.8%); p<0.05). The mean (SD) heart rate during total time was 171 (4) beats/min, with a significant difference (p<0.01) between guards and centres. Mean (SD) plasma lactate concentration was 5.49 (1.24) mmol/l, with concentrations at half time (6.05 (1.27) mmol/l) being significantly (p<0.001) higher than those at full time (4.94 (1.46) mmol/l). The changes to the rules of basketball have slightly increased the cardiac efforts involved during competition. The game intensity may differ according to the playing position, being greatest in guards. PMID- 17138631 TI - A 6-month prospective study of injury in Gaelic football. AB - OBJECTIVE: To describe the injury incidence in Gaelic football. METHODS: A total of 83 players from three counties were interviewed monthly about their injury experience, during the 6 months of the playing season. RESULTS: The injury rate was 13.5/1000 h exposure to Gaelic football (95% CI, 10.9 to 16.6). There were nearly twice as many injuries during matches (64.4%, 95% CI, 54.1 to 73.6) as in training (35.6%, 95% CI, 26.4 to 49.5). The ankle was found to be the most commonly injured site (13.3%, 95% CI, 7.8 to 21.9). The musculotendinous unit accounted for nearly 1/3 of all injuries (31.1%). The tackle accounted for 27.8% of the injuries sustained (tackler 10%, 95% CI, 5.4 to 17.9; player being tackled 17.9%, 95% CI, 11.2 to 26.9). Of total match injuries, 56.9% (95% CI, 46.1 to 67.1) were experienced in the second half as opposed to 39.7% (95% CI, 29.8 to 50.5) in the first half. CONCLUSIONS: Gaelic footballers are under considerable risk of injury. Greater efforts must be made to reduce this risk so that players miss less time from sport due to injury. Risk factors for injury in Gaelic football must now be investigated so that specific interventions may be established to reduce them. PMID- 17138632 TI - Effect of off-road competitive motocross race on plasma oxidative stress and damage markers. AB - AIM: To analyse the effect of an off-road motocross heat on plasma levels of oxidative stress and damage, blood leucocyte counts and urine catecholamine concentration. METHODS: Plasma contents of total, reduced and oxidised (GSSG) glutathione, %GSSG, malondialdehyde (MDA), protein carbonyl and sulphydryl groups, total antioxidant status (TAS), uric acid, and blood neutrophil and lymphocyte counts were evaluated in 10 male top-level riders before, immediately after (0 h) and 1 h after a simulated competitive motocross race. 24-h urine adrenaline, noradrenaline and dopamine concentrations were also measured. RESULTS: The motocross heat resulted in an increase in plasma oxidative stress and damage (p<0.05). This was shown by a significant increase in %GSSG, TAS, MDA and carbonyls, and by a decrease in sulphydryl groups after the race. There was a significant increase in both plasma uric acid and urine catecholamine concentration after the race (p<0.05). Blood neutrophil counts increased at 0 and 1 h after exercise (p<0.05). Lymphocyte count increased from baseline to 0 h, although it decreased from baseline and 0 to 1 h after exercise (p<0.05). CONCLUSION: The data reinforce the marked metabolic and hormonal demands imposed by motocross, resulting in a condition of enhanced plasma oxidative stress and damage. PMID- 17138633 TI - A study on tinea gladiatorum in young wrestlers and dermatophyte contamination of wrestling mats from Sari, Iran. AB - OBJECTIVE: To study the prevalence of tinea gladiatorum among young wrestlers and dermatophyte contamination of wrestling mats from Sari city, the capital of Mazandaran, a northern province of Iran. DESIGN: 324 wrestlers (aged 9-20 years) from 7 active clubs in Sari city were examined, and skin scrapings were obtained from 135 wrestlers suspected of having tinea gladiatorum. The scraped skin samples were evaluated with potassium hydroxide. Pleated carpet sterile fragments (5 x 5 cm) were used for to survey of wrestling mat contamination. Sabouraud's dextrose agar with and without chloramphenicole and cyclohexamide was used to culture scrapings and wrestling mat samples. The dermatophytes were identified by routine laboratory techniques. RESULTS: Our study showed that of the 324 wrestlers, 65 (20.1%) had tinea gladiatorum. Most lesions were on the trunk and head. All the wrestling mat samples were positive for dermatophytes. Trichophyton tonsurans was isolated from all the scrapings and wrestling mat samples. CONCLUSION: Considering that several colonies of T tonsurans were isolated from all the wrestling mats and from wrestlers with tinea gladiatorum (as the only dermatophyte species) we think that the contamination of wrestling mats with T tonsurans has a crucial role in the injection of wrestlers. PMID- 17138634 TI - Effect of maximal exercise on the short-term kinetics of zinc metabolism in sedentary men. AB - OBJECTIVE: To examine the effect of acute exhaustive exercise versus rest on short-term zinc kinetics in men. DESIGN: Crossover design, wherein all subjects were their own control. SETTING: University setting, where subjects were free living. PARTICIPANTS: 12 healthy, sedentary men, 25-35 years of age. INTERVENTIONS: (70)Zn was infused 10 min after exercise or at rest. Plasma zinc concentrations were measured at baseline and 2, 5, 10, 15, 30, 45, 60, 75, 90 and 120 min after exercise or rest. Haematocrit was measured before and after exercise to assess changes in plasma volume. MAIN OUTCOME MEASUREMENTS: Plasma zinc (primary), serum creatine kinase and serum cortisol concentrations (secondary). RESULTS: Plasma zinc concentrations decreased (p<0.05) after exercise, with a mean (SD) nadir of 13.9% (4.1%) observed at 70 min after exercise. There were increases in the size of the rapidly exchangeable plasma zinc pool (Qa; from 3.1 (0.2) to 3.6 (0.2) mg; p<0.05) and the liver zinc pool (Qb; from 10.2 (0.6) to 11.4 (0.8) mg; p = 0.12). CONCLUSION: Exercise seems to cause a shift of plasma zinc into the interstitial fluid and liver after exercise, which may reflect the acute stress response of strenuous exercise. PMID- 17138636 TI - Intermittent KoldBlue cryotherapy of 3x10 min changes mid-portion Achilles tendon microcirculation. AB - BACKGROUND: Neovascularisation and microcirculatory changes have been reported in Achilles tendinopathy. Cryotherapy and compression, as part of a rest, ice, compression and elevation regimen, are shown to decrease pain and improve function. However, the microcirculatory changes following a given dosage of cryotherapy on mid-portion Achilles tendon remain unclear. STUDY DESIGN: Prospective clinical cohort study, level of evidence 2. METHODS: 30 people (12 males, 33 (SD 12) years, body mass index 25.6 (5.3) kg/m2) were included in the cohort. 3x10 min KoldBlue ankle-cooling bandages were applied and microcirculation of Achilles tendon mid-portion was real-time and continuously assessed using a laser-Doppler-spectrophotometry system (O2C, Germany). RESULTS: Superficial capillary blood flow was reduced from 42 to 6, 5 and 3 relative units (rU) in the first, second and third cryotherapy periods, respectively (-65%, p = 0.001), with no significant capillary hyperaemia. Deep capillary tendon blood flow was reduced from 180 to 82, 53 and 52 rU (-71%, p = 0.001) within 6-9 min of application without hyperaemia. Superficial tendon oxygen saturation dropped significantly from 43% to 26%, 18% and 11% (p = 0.001) after repetitive cryotherapy, with persisting increase of tendon oxygenation during rewarming (51%, 49% and 54%, p = 0.077) up to 27% of the baseline level. At 8 mm tendon depth, cryotherapy preserved local oxygenation. Relative postcapillary venous tendon filling pressures were favourably reduced from 41 (11) to 31, 28 and 26 rU (-36%, p = 0.001) superficially and from 56 (11) to 45, 46 and 48 rU (-18%, p = 0.001) in deep capillary blood flow during cryotherapy, facilitating capillary venous clearance. CONCLUSION: Intermittent cryotherapy of 3x10 min significantly decreases local Achilles tendon mid-portion capillary blood flow by 71%. Within 2 min of rewarming, tendon oxygen saturation is re-established following cryotherapy. Postcapillary venous filling pressures are reduced during cryotherapy, favouring capillary venous outflow of the healthy Achilles tendon. PMID- 17138635 TI - Influence of vibration on delayed onset of muscle soreness following eccentric exercise. AB - Delayed onset muscle soreness (DOMS), which may occur after eccentric exercise, may cause some reduction in ability in sport activities. For this reason, several studies have been designed on preventing and controlling DOMS. As vibration training (VT) may improve muscle performance, we designed this study to investigate the effect of VT on controlling and preventing DOMS after eccentric exercise. METHODS: Fifty healthy non-athletic volunteers were assigned randomly into two experimental, VT (n = 25) and non-VT (n = 25) groups. A vibrator was used to apply 50 Hz vibration on the left and right quadriceps, hamstring and calf muscles for 1 min in the VT group, while no vibration was applied in the non VT group. Then, both groups walked downhill on a 10 degrees declined treadmill at a speed of 4 km/hour. The measurements included the isometric maximum voluntary contraction force (IMVC) of left and right quadriceps muscles, pressure pain threshold (PPT) 5, 10 and 15 cm above the patella and mid-line of the calf muscles of both lower limbs before and the day after treadmill walking. After 24 hours, the serum levels of creatine-kinase (CK), and DOMS level by visual analogue scale were measured. RESULTS: The results showed decreased IMVC force (P = 0.006), reduced PPT (P = 0.0001) and significantly increased mean of DOMS and CK levels in the non-VT group, compared to the VT group (P = 0.001). CONCLUSION: A comparison by experimental groups indicates that VT before eccentric exercise may prevent and control DOMS. Further studies should be undertaken to ascertain the stability and effectiveness of VT in athletics. PMID- 17138637 TI - Syncope due to Brugada syndrome in a young athlete. AB - A 30-year-old male athlete with exercise-related syncopal symptoms spontaneously exhibited a type 1 Brugada ECG and was inducible during electrophysiology study. He was diagnosed with symptomatic Brugada syndrome and deemed at high risk of sudden cardiac death. Thus, he received a cardioverter/defibrillator and was advised to abstain from further competitive sports activities. This case points to a role of the ECG in pre-participation screening. It also demonstrates that, in athletes with Brugada syndrome, repolarisation anomalies may be markedly attenuated during vigorous exercise and considerably increased immediately after exercise. The observed J-wave amplitude dynamics suggests enhancement of pre existing autonomic dysfunction through heavy exertion. PMID- 17138638 TI - Hip arthroscopy: current concepts and review of literature. AB - Diagnosis and treatment of intra-articular hip problems in young patients present a challenge to hip surgeons. Previous studies have shown that non-invasive investigations such as radiography, computed tomography and magnetic resonance imaging provide limited help. Non-operative treatment is likely to result in persistent symptoms, and surgical options for intra-articular hip problems involve open arthrotomy of the hip joint, which carries potential risks associated with joint dislocation. Arthroscopy of the hip joint, therefore, seems to be an attractive option. It was once thought that introduction of a straight arthroscope into the ball-and-socket hip joint was almost impossible. Hip arthroscopy has seen several advances since then, and the speed at which it developed in recent years directly corresponded to the rate at which the conditions affecting the hip joint were identified. Athletes and other young individuals with hip injuries are increasingly being diagnosed with an ever evolving series of conditions. Many of these conditions were previously unrecognised and thus left untreated, resulting in premature ends to the patients' competitive careers. Hip arthroscopy, as with any procedure, is not without risks. The procedure is not widely available as it requires specialist equipment and takes a long time to learn. Complications are few, occurring in <5% of patients. PMID- 17138639 TI - Difference in plantar pressure between the preferred and non-preferred feet in four soccer-related movements. AB - OBJECTIVE AND PARTICIPANTS: The present study measured the difference in plantar pressure between the preferred and non-preferred foot in four soccer-related movements in 15 male university soccer players (mean (SD) age 20.9 (1.3) years, mean (SD) height 173 (4) cm and mean (SD) weight 61.7 (3.6) kg). DESIGN: To record plantar pressure distribution, players randomly wore three types of soccer shoes (classical 6-stud and 12-stud, and specially designed 12-stud) embedded with an insole pressure recorder device with 99 sensors, divided into 10 areas for analysis. Plantar pressure was recorded in five successful trials in each of the four soccer-related movements: running (at 3.3 m/s), sideward cutting, 45 degrees cutting and landing from a vertical jump. RESULTS: Plantar pressures of the preferred and non-preferred foot were different in 115 of 120 comparisons. The overall plantar pressure of the preferred foot was higher than that of the non-preferred foot. Specifically, in each of the four movements, higher pressure was found in the preferred foot during the take-off phase, whereas this was found in the non-preferred foot during the landing phase. This would suggest a tendency of the preferred foot for higher motion force and of the non-preferred foot for a greater role in body stabilisation. CONCLUSIONS: The data indicate that the preferred and non-preferred foot should be treated independently with regard to strength/power training to avoid unnecessary injuries. Different shoes/insoles and different muscular strengthening programmes are thus suggested for each of the soccer player's feet. PMID- 17138640 TI - The "bench-presser's shoulder": an overuse insertional tendinopathy of the pectoralis minor muscle. AB - BACKGROUND: Tendinopathies of the rotator cuff muscles, biceps tendon and pectoralis major muscle are common causes of shoulder pain in athletes. Overuse insertional tendinopathy of pectoralis minor is a previously undescribed cause of shoulder pain in weightlifters/sportsmen. OBJECTIVES: To describe the clinical features, diagnostic tests and results of an overuse insertional tendinopathy of the pectoralis minor muscle. To also present a new technique of ultrasonographic evaluation and injection of the pectoralis minor muscle/tendon based on use of standard anatomical landmarks (subscapularis, coracoid process and axillary artery) as stepwise reference points for ultrasonographic orientation. METHODS: Between 2005 and 2006, seven sportsmen presenting with this condition were diagnosed and treated at the Cape Shoulder Institute, Cape Town, South Africa. RESULTS: In five patients, the initiating and aggravating factor was performance of the bench-press exercise (hence the term "bench-presser's shoulder"). Medial juxta-coracoid tenderness, a painful active-contraction test and bench-press manoeuvre, and decrease in pain after ultrasound-guided injection of a local anaesthetic agent into the enthesis, in the absence of any other clinically/radiologically apparent pathology, were diagnostic of pectoralis minor insertional tendinopathy. All seven patients were successfully treated with a single ultrasound-guided injection of a corticosteroid into the enthesis of pectoralis minor followed by a period of rest and stretching exercises. CONCLUSIONS: This study describes the clinical features and management of pectoralis minor insertional tendinopathy, secondary to the bench-press type of weightlifting. A new pain site-based classification of shoulder pathology in weightlifters is suggested. PMID- 17138641 TI - Haemodynamic changes induced by submaximal exercise before a dive and its consequences on bubble formation. AB - OBJECTIVES: To evaluate the effects of a submaximal exercise performed 2 h before a simulated dive on bubble formation and to observe the haemodynamic changes and their influence on bubble formation. PARTICIPANTS AND METHODS: 16 trained divers were compressed in a hyperbaric chamber to 400 kPa for 30 min and decompressed at a rate of 100 kPa/min with a 9 min stop at 130 kPa (French Navy MN90 procedure). Each diver performed two dives 3 days apart, one without exercise and one with exercise before the dive. All participants performed a 40 min constant-load submaximal and calibrated exercise, which consisted of outdoor running 2 h before the dive. Circulating bubbles were detected with a precordial Doppler at 30, 60 and 90 min after surfacing. Haemodynamic changes were evaluated with Doppler echocardiography. RESULTS: A single bout of strenuous exercise 2 h before a simulated dive significantly reduced circulating bubbles. Post-exercise hypotension (PEH) was observed after exercise with reductions in diastolic and mean blood pressure (DBP and MBP), but total peripheral resistance was unchanged. Stroke volume was reduced, whereas cardiac output was unchanged. Simulated diving caused a similar reduction in cardiac output independent of pre-dive exercise, suggesting that pre-dive exercise only changed DBP and MBP caused by reduced stroke volume. CONCLUSION: A single bout of strenuous exercise 2 h before a dive significantly reduced the number of bubbles in the right heart of divers and protected them from decompression sickness. Declining stroke volume and moderate dehydration induced by a pre-dive exercise might influence inert gas load and bubble formation. PMID- 17138642 TI - Topical ketoprofen TDS patch versus diclofenac gel: efficacy and tolerability in benign sport related soft-tissue injuries. AB - OBJECTIVE: To compare the ketoprofen TDS patch with diclofenac gel in the treatment of traumatic acute pain in benign sport-related soft-tissue injuries. DESIGN: 7-14 treatment days, prospective, randomised, open study. PATIENTS: Outpatients aged 18-70 years diagnosed for painful benign sport-related soft tissue injury (sprains, strains and contusions within the prior 48 h), randomised to either ketoprofen patch 100 mg once daily (n = 114) or diclofenac gel 2-4 g three times daily (n = 109). INTERVENTION: 7-14 days of topical non-steroidal anti-inflammatory drugs treatment to assess the pain intensity changes (daily activities and spontaneous at rest) in a daily diary (100-mm Visual Analogue Scale (VAS)). MAIN OUTCOME MEASUREMENT: Pain intensity (VAS). RESULTS: The ketoprofen patch was not inferior to diclofenac gel in reducing the baseline pain during daily activities (difference of -1.17 mm in favour of ketoprofen patch, 95% CI (-5.86 to 3.52), reducing to the baseline VAS 79%. Ketoprofen patch presented also a higher cure rate (64%) than diclofenac gel (46%) at day 7 (p = 0.004). Patient opinions about the treatment comfort (pharmaceutical shape, application and dosage) were also statistically higher for the ketoprofen patch (>80% of the patients rated as good or excellent the patch removal and skin adherence). CONCLUSION: Ketoprofen patches are effective and safe pain relievers for the treatment of sports injury pain with advantages compared with diclofenac gel. PMID- 17138643 TI - A stress fracture of the lumbar spine in a professional rugby player. AB - In modern rugby, the spine is subjected to great physical pressure, with an increased number of impacts, on both the cervical and the lumbar spine. This case report illustrates overuse injuries of the lumbar spine in international professional rugby players. A 32-year-old sportsman had been practising rugby for 24 years and was playing for a championship level French team when he started suffering from a right lateral pain in the lumbosacral spine. A CT scan showed a unilateral isthmolysis and a coronal irregular fracture of the right pars interarticularis of L5. This led to discussion of (1) the importance of the decision on the date of return to playing rugby and (2) the future of professional rugby players with chronic spinal injuries. PMID- 17138644 TI - The role of extranuclear signaling actions of progesterone receptor in mediating progesterone regulation of gene expression and the cell cycle. AB - Human progesterone receptor (PR) contains a motif that interacts with the SH3 domain of Src and mediates rapid activation of Src and downstream MAPK (Erk-1/-2) without relying on the transcriptional activity of the receptor. Here we investigated the role and intracellular location of this nontranscriptional activity of PR. Progestin activation of Src/MAPK occurred outside the nucleus with the B isoform of PR that was distributed between the cytoplasm and nucleus, but not with PR-A that was predominantly nuclear. Breast cancer cells stably expressing wild-type PR-B or PR-B with disrupting point mutations in the SH3 domain binding motif (PR-BDeltaSH3) that do not affect the transcriptional activity of PR, were compared for effects of progestin on endogenous target gene expression and cell proliferation. Progestin induction of the cyclin D1 gene, which lacks a progesterone response element, was dependent on PR activation of the Src/MAPK pathway, whereas induction of the Sgk (serum and glucocorticoid regulated kinase) gene that contains a functional progesterone response element was unaffected by mutations that interfere with PR activation of Src. Progestin induction of cell cycle progression was also abrogated in cells expressing PR BDeltaSH3, and no effect of progestin on cyclin D1 expression and cell cycle was observed in the presence of PR-A. These results highlight the importance of PR activation of the Src/MAPK signaling pathway for progesterone-induced transcription of select target genes and cell cycle progression. PMID- 17138645 TI - Reciprocal cross talk between gonadotropin-releasing hormone (GnRH) and prostaglandin receptors regulates GnRH receptor expression and differential gonadotropin secretion. AB - The asynchronous secretion of gonadotrope LH and FSH under the control of GnRH is crucial for ovarian cyclicity but the underlying mechanism is not fully resolved. Because prostaglandins (PG) are autocrine regulators in many tissues, we determined whether they have this role in gonadotropes. We first demonstrated that GnRH stimulates PG synthesis by induction of cyclooxygenase-2, via the protein kinase C/c-Src/phosphatidylinositol 3'-kinase/MAPK pathway in the LbetaT2 gonadotrope cell line. We then demonstrated that PGF(2alpha) and PGI2, but not PGE2 inhibited GnRH receptor expression by inhibition of phosphoinositide turnover. PGF(2alpha), but not PGI2 or PGE2, reduced GnRH-induction of LHbeta gene expression, but not the alpha-gonadotropin subunit or the FSHbeta subunit genes. The prostanoid receptors EP1, EP2, FP, and IP were expressed in rat gonadotropes. Incubations of rat pituitaries with PGF(2alpha), but not PGI2 or PGE2, inhibited GnRH-induced LH secretion, whereas the cyclooxygenase inhibitor, indomethacin, stimulated GnRH-induced LH secretion. None of these treatments had any effect on GnRH-induced FSH secretion. The findings have thus elaborated a novel GnRH signaling pathway mediated by PGF(2alpha)-FP and PGI2-IP, which acts through an autocrine/paracrine modality to limit autoregulation of the GnRH receptor and differentially inhibit LH and FSH release. These findings provide a mechanism for asynchronous LH and FSH secretions and suggest the use of combination therapies of GnRH and prostanoid analogs to treat infertility, diseases with unbalanced LH and FSH secretion and in hormone-dependent diseases such as prostatic cancer. PMID- 17138646 TI - MAPK signaling is critical to estradiol protection of CA1 neurons in global ischemia. AB - The importance of hormone therapy in affording protection against the sequelae of global ischemia in postmenopausal women remains controversial. Global ischemia arising during cardiac arrest or cardiac surgery causes highly selective, delayed death of hippocampal CA1 neurons. Exogenous estradiol ameliorates global ischemia induced neuronal death and cognitive impairment in male and female rodents. However, the molecular mechanisms by which estrogens intervene in global ischemia induced apoptotic cell death are unclear. Here we show that estradiol acts via the classical estrogen receptors, the IGF-I receptor, and the ERK/MAPK signaling cascade to protect CA1 neurons in ovariectomized female rats and gerbils. We demonstrate that global ischemia promotes early dephosphorylation and inactivation of ERK1 and the transcription factor cAMP-response element binding protein (CREB), subsequent down-regulation of the antiapoptotic protein Bcl-2, a known gene target of estradiol and CREB, and activation of caspase-3. Estradiol treatment increases basal phosphorylation of both ERK1 and ERK2 in hippocampal CA1 and prevents ischemia-induced dephosphorylation and inactivation of ERK1 and CREB, down-regulation of Bcl-2 and activation of the caspase death cascade. Whereas ERK/MAPK signaling is critical to CREB activation and neuronal survival, the impact of estradiol on Bcl-2 levels is ERK independent. These findings support a model whereby estradiol acts via the classical estrogen receptors and IGF-I receptors, which converge on activation of ERK/MAPK signaling and CREB to promote neuronal survival in the face of global ischemia. PMID- 17138647 TI - Hypotonicity induces aquaporin-2 internalization and cytosol-to-membrane translocation of ICln in renal cells. AB - Kidney collecting-duct cells swell in response to changes in medulla osmolality caused by the transition from antidiuresis to diuresis. Regulatory volume decrease (RVD) mechanisms must be activated to face this hypotonic stress. In Aquaporin-2 (AQP2)-expressing renal CD8 cells, hypotonicity decreased cell surface expression of AQP2 and increased the amount of AQP2 localized intracellularly, whereas the total amount of AQP2 phosphorylated at ser-256 decreased. Analysis of cAMP dynamics using fluorescence resonance energy transfer (FRET) showed that hypotonicity causes a reduction of cAMP, consistent with a decrease in phospho-AQP2. Moreover, hypotonicity caused a profound actin reorganization, associated with the loss of stress fibers and formation of F actin patches (microspikes) at the cell border. Those changes were regulated by the monomeric GTPase Cdc42. Interestingly, expression of the dominant-negative Cdc42 (N17-Cdc42) prevented the hypotonicity-induced microspike formation and the generation of Cl(-) currents. Hypotonicity also caused the relocation from the cytosol to the plasma membrane and increase in interaction with actin of ICln (nucleotide-sensitive chloride current protein), which is essential for the generation of ion currents activated during RVD. Together, the profound actin remodeling, internalization of AQP2 and translocation of ICln to the plasma membrane during hypotonicity may contribute to RVD after cell swelling in renal medulla. PMID- 17138648 TI - Posterior Hox gene expression and differential androgen regulation in the developing and adult rat prostate lobes. AB - Axis positioning and tissue determination during development involve coordinated expression of Hox genes throughout the body. The most posterior Hox gene clusters are involved in prostate organogenesis. In the present study, we characterized and compared the expression profiles of posterior (5') Hox genes in the separate lobes of the adult rat prostate gland, the coagulating gland, seminal vesicles, and epididymis using quantitative real-time RT-PCR. These genes include Hoxa9-11, Hoxa13, Hoxd13, and Hoxb13. We identified a unique Hox code for each of these organs and propose that this contributes to the organ-specific and prostate lobe specific identities in the adult rat. Using the ventral prostate (VP) as a model, we characterized the Hox genes expression patterns over time from birth through adulthood. Expression levels of the three Hox13 genes and Hoxa10 were significantly higher in the adult VP compared with the neonatal developing VP suggesting an important role during adult homeostasis. In contrast, Hoxa9 and Hoxa11 levels declined after morphogenesis suggesting a specific developmental role. Overall, the Hoxb13 gene exhibited the most striking temporal and organ specific differences. Using in situ hybridization and immunohistochemistry, a distinct Hoxb13 anterior-to-posterior expression gradient was observed with the highest expression levels in the VP luminal epithelial cells, moderate levels in the lateral prostate, and low expression in the dorsal prostate. An expression gradient was also observed along the ductal length in all three prostate lobes with strongest expression at the distal tips and limited expression in the proximal ducts. After infection with a lentivirus expressing the Hoxb13 gene, NRP 152 cells cultured under nondifferentiating conditions exhibited robust cytokeratin 8 immunostain indicating that Hoxb13 expression drives luminal cell differentiation in the rat epithelium. Androgen regulation of prostatic Hox gene expression was examined during development in vitro and after castration in the adult rat. In the neonatal VP, all six Hox genes were significantly up-regulated by androgens, whereas none of the genes were affected by testosterone in the lateral prostate. In the adult rat, castration resulted in up-regulation of Hoxa9 and Hoxa13 in the VP and down-regulation of Hoxb13 in the dorsal prostate and lateral prostate. Taken together, we conclude that the prostatic Hox genes reach a destined expression level at specific developmental time points in the prostate gland and possess differential androgenic regulation in a temporal and lobe specific manner. We suggest that this timely Hox code participates in determining lobe-specific prostatic identity and cellular differentiation. PMID- 17138650 TI - Transient transcription of the somatostatin gene at the time of estrogen dependent organization of the sexually dimorphic nucleus of the rat preoptic area. AB - In situ hybridization detected a transient, sex-specific transcription of somatostatin gene in the central part of the rat medial preoptic nucleus, which coincides with the sexually dimorphic nucleus of the preoptic area (SDN-POA), during, but not after, the establishment of sex difference. On postnatal d 1 (day of birth), somatostatin mRNA was detected in the SDN-POA of both sexes. On d 8 through 35, the area of somatostatin mRNA-positive cells was significantly larger in males than in females. In males the area attained its maximum size on d 15 and diminished gradually thereafter. In females the area did not change in size during this period. On d 60 expression of somatostatin mRNA was low and not different between sexes. Throughout the observed period, Nissl staining and calbindin immunohistochemistry enabled visualization of the typical SDN-POA in the same region. As with Nissl staining and calbindin immunohistochemistry, somatostatin mRNA hybridization on d 15 revealed a reversal of the sexual dimorphism in the size of the SDN-POA in males that had been neonatally orchidectomized or females given estrogen as pups, showing that sex steroid milieu during the organizational period determines the area occupied by somatostatin mRNA-positive cells. Sex-specific, transient transcription of the somatostatin gene may causally relate to the estrogen-dependent organization of the SDN-POA. PMID- 17138649 TI - In vivo response-based identification of direct hormone target cell populations using high-density tissue arrays. AB - To identify cell populations directly responsive to prolactin (PRL), GH, erythropoietin, or granulocyte-colony stimulating factor within the physiological setting of an intact mammal, we combined in situ detection of hormone-activated signal transducer and activator of transcription (Stat)-5 in rats with high throughput tissue array analysis using cutting-edge matrix assembly (CEMA). Inducible activation of Stat5a/b, as judged by levels of nuclear-localized, phosphoTyr694/699-Stat5a/b, served as an immediate and sensitive in situ marker of receptor signaling in rat tissues after injection into male and female rats of a single, receptor-saturating dose of hormone for maximal receptor activation. CEMA tissue arrays facilitated analysis of most tissues, including architecturally complex, thin-walled, and stratified tissues such as gut and skin. In 40 tissues analyzed, 35 PRL-responsive and 32 GH-responsive cell types were detected, of which 22 cell types were responsive to both hormones. Interestingly, PRL but not GH activated Stat5 in nearly all of the endocrine glands. In mammary glands, PRL activated Stat5 in a majority of luminal epithelial cells but not myoepithelial cells, stromal fibroblasts, or adipocytes, whereas GH activated Stat5 in a significant fraction of myoepithelial cells, fibroblasts, and adipocytes but only in a minority of luminal cells. Finally, the organism-wide screening revealed a yet-to-be identified erythropoietin-responsive cell type in connective tissue. CEMA tissue arrays provide cost-effective in situ analysis of large numbers of tissues. Biomarker-based identification of cell populations responsive to individual hormones may shed new light on endocrine disease as well as improve understanding of effects and side effects of hormones and drugs. PMID- 17138651 TI - Ontogeny and effects of hypothalamic pituitary disconnection on formation of inositol trisphosphate in fetal sheep pituitary cells. AB - In late gestation fetal sheep, the pituitary becomes increasingly responsive to stimulation by arginine vasopressin (AVP). This change appears to be one important factor mediating the plasma cortisol surge, a critical developmental event. It is not known precisely why pituitary corticotropes become more responsive at this time. In this study we examined the possibility that changes in second messenger generation [inositol trisphosphate (IP(3))] are responsible. Two studies were undertaken. The first was an ontogeny study, where pituitaries were isolated from 100-, 120-, and 140-d gestational age (dGA) fetal sheep. Cells were cultured, stimulated with AVP, and the formation of IP(3) assessed. The amount of IP(3) generated increased with gestational age (percent increases from unstimulated controls were 4.6, 11.5, and 21.5 for 100, 120, and 140 dGA, respectively), with significant differences between the 140-dGA group and both earlier groups apparent. The second study examined the impact of 120-dGA hypothalamo-pituitary disconnection (HPD), which prevents corticotrope maturation, on responsiveness of pituitary cells isolated from 140-dGA fetuses. Cells were stimulated with AVP, and the formation of IP(3) and secretion of ACTH were assessed. Significantly less IP(3) was formed, and ACTH secreted in cells from HPD compared with control fetuses (IP(3) and ACTH levels were 50% and 35% lower, respectively). Results from the HPD study demonstrate that the ontogenic changes in IP(3) after AVP require an intact hypothalamic-pituitary-adrenal axis. These findings suggest that heightened second messenger generation may be a key reason for increased ACTH secretory responsiveness to AVP in the late gestation sheep fetus. PMID- 17138652 TI - The effect of estradiol on in vivo tumorigenesis is modulated by the human epidermal growth factor receptor 2/phosphatidylinositol 3-kinase/Akt1 pathway. AB - To determine whether the epidermal growth factor receptor 2 (ErbB2) and Akt1 can alter the in vivo growth of MCF-7 cells, parental cells or cells stably transfected with constitutively active Akt1 (myr-Akt1) or dominant-negative Akt1 mutants (K179M-Akt1 and R25C-Akt1) were implanted into athymic nude mice. Tumor growth was monitored in the presence or absence of the antiestrogen tamoxifen and the selective ErbB2 inhibitor, AG825. MCF-7 [parental or empty vector transfected, cytomegalovirus (CMV)] and myr-Akt1 cells formed tumors upon estradiol supplementation after 20-30 d (59-, 29-, and 17-fold increase in tumor volume, respectively). Tamoxifen and AG825 blocked the estradiol effect by 93 and 96% in MCF-7 xenografts, 88 and 81% in CMV xenografts, and 91% in myr-Akt1 xenografts. Furthermore, AG825 suppressed the growth of established tumors in CMV and myr-Akt1 inoculated animals by 68 and 75%, respectively, as compared with continued estrogen supplementation, suggesting a role for ErbB2. When K179M-Akt1 or R25C-Akt1 cells were injected into ovariectomized animals, tumor growth was reduced upon estradiol treatment by 95% and 98%, respectively, supporting a role for Akt1. In contrast to ovariectomized animals, in intact animals, myr-Akt1 cells could establish tumors without estradiol priming after 40-50 d (20-fold increase in tumor volume). Loss of Akt1 phosphorylation was associated with tumor growth inhibition. Immunohistochemical assays showed that in tumors from parental and CMV xenografts, estradiol decreased estrogen receptor-alpha expression and induced progesterone receptor expression and Akt phosphorylation, effects that were inhibited by tamoxifen, AG825, and R25C-Akt1 by 89, 82, and 77% for progesterone receptor expression and 48, 66, and 73% for pAkt expression, respectively. Cumulatively, our results suggest that Akt1 and ErbB2 are involved in in vivo tumorigenesis and modulation of estrogen receptor-alpha expression and activity. PMID- 17138653 TI - Characterization and functional role of androgen-dependent PDE5 activity in the bladder. AB - Benign prostate hyperplasia is the most common disease in the aging male, often comorbid with erectile dysfunction. Phosphodiesterase type 5 (PDE5) inhibitors (sildenafil, tadalafil, and vardenafil) decrease lower urinary tract symptoms in patients with erectile dysfunction and BPH. We studied PDE5 expression and activity in the human bladder and PDE5i effects both in vitro (human and rat) and in vivo (rat). PDE5 is highly expressed in rat and human bladder and immunolocalized in vascular endothelium and muscle fibers. Sildenafil, tadalafil, and vardenafil blocked 70% of the total cGMP-catabolizing activity; vardenafil was the most potent (IC(50) = 0.3 nm). In human bladder cells and in rat strips, a PDE-resistant cGMP analog, SP-8-Br-PET-cGMPS, induced, respectively, a consistent antiproliferative and relaxant effect. In contrast, the nitric oxide donor sodium nitroprusside (SNP) was almost ineffective. However, blocking PDE5 with vardenafil increased SNP antiproliferative and relaxant activity up to the level observed with SP-8-Br-PET-cGMPS. We also found that castration decreased, and T supplementation restored, PDE5 gene expression in rat bladder. Accordingly, bladder strips from castrated rats were more sensitive to SNP-induced relaxation than strips from control or T-replaced rats, whereas in the presence of vardenafil, all groups showed the same SNP sensitivity. To discover whether vardenafil affects bladder activity in vivo, the rat bladder outlet obstruction model was used. Chronic treatment with 10 mg/kg.d vardenafil significantly reduced nonvoiding contractions (47%, P < 0.05 vs. placebo) up to tamsulosin level (51%). Overall, these results demonstrate that PDE5 regulates bladder smooth muscle tone, strongly limiting the nitric oxide/cGMP signaling, and that vardenafil, by blocking PDE5, may be a possible therapeutic option for bladder dysfunction by ameliorating irritative lower urinary tract symptoms. PMID- 17138654 TI - Mice with impaired extrathyroidal thyroxine to 3,5,3'-triiodothyronine conversion maintain normal serum 3,5,3'-triiodothyronine concentrations. AB - For T(3) to mediate its biological effects, the prohormone T(4) must be activated by removal of an outer-ring iodine by the type 1 or 2 deiodinases (D1 and D2) with approximately 60% of the daily T(3) production in rodents being produced extrathyroidally through this pathway. To further define the role of these enzymes in thyroid hormone homeostasis, we backcrossed the targeted disruption of the Dio2 gene into C3H/HeJ (C3H) mice with genetically low D1 expression to create the C3H-D2KO mouse. Remarkably, these mice maintain euthyroid serum T(3) levels with normal growth and no decrease in expression of hepatic T(3) responsive genes. However, serum T(4) is increased 1.2-fold relative to the already elevated C3H levels, and serum TSH is increased 1.4-fold. Despite these increases, thyroidal (125)I uptake indicates no difference in thyroidal activity between C3H-D2KO and C3H mice. Although C3H-D2KO hepatic and renal D1 activities were well below those observed in wild-type mice (approximately 0.1-fold for both), they were 8-fold and 2-fold higher, respectively, relative to C3H mice. Thyroidal D1 and cerebral cortical type 3 deiodinase activity were unchanged between C3H-D2KO and C3H mice. In conclusion, C3H-D2KO mice have notably elevated serum T(4) levels, and this, in conjunction with residual D1 activity, is likely an important role in the maintenance of euthyroid serum T(3) concentrations. PMID- 17138655 TI - A growth hormone-releasing peptide promotes mitochondrial biogenesis and a fat burning-like phenotype through scavenger receptor CD36 in white adipocytes. AB - Whereas the uptake of oxidized lipoproteins by scavenger receptor CD36 in macrophages has been associated with foam cell formation and atherogenesis, little is known about the role of CD36 in regulating lipid metabolism in adipocytes. Here we report that treatment of 3T3-L1 adipocytes with hexarelin, a GH-releasing peptide that interacts with CD36, resulted in a depletion of intracellular lipid content with no significant change in CD36 expression. Microarray analysis revealed an increased pattern in several genes involved in fatty acid mobilization toward the mitochondrial oxidative phosphorylation process in response to hexarelin. Interestingly, many of these up-regulated genes are known targets of peroxisomal proliferator-activated receptor (PPAR)-gamma, such as FATP, CPT-1, and F(1)-ATPase, suggesting that adipocyte response to hexarelin may involve PPARgamma activation. Expression studies also indicate an increase in thermogenic markers PPARgamma coactivator 1alpha and uncoupling protein-1, which are normally expressed in brown adipocytes. Electron microscopy of hexarelin-treated 3T3-L1 adipocytes showed an intense and highly organized cristae formation that spans the entire width of mitochondria, compared with untreated cells, and cytochrome c oxidase activity was enhanced by hexarelin, two features characteristic of highly oxidative tissues. A similar mitochondrial phenotype was detected in epididymal white fat of mice treated with hexarelin, along with an increased expression of thermogenic markers that was lost in treated CD36-null mice, suggesting that the ability of hexarelin to promote a brown fat-like phenotype also occurs in vivo and is dependent on CD36. These results provide a potential role for CD36 to impact the overall metabolic activity of fat usage and mitochondrial biogenesis in adipocytes. PMID- 17138656 TI - Bone morphogenetic protein and retinoic acid-inducible neural specific protein-3 is expressed in gonadotrope cell pituitary adenomas and induces proliferation, migration, and invasion. AB - Pituitary tumors are common intracranial neoplasms that often result in endocrine dysfunction due to hormone overproduction or deficiencies from mass effects. Gonadotrope cell or gonadotropinomas are tumors that produce LH and/or FSH and represent 40% of macroadenomas. Little is known about their underlying pathogenic mechanisms. We compared expression profiles of 10 gonadotropinomas with nine normal pituitaries by cDNA array and identified bone morphogenetic protein- and retinoic acid-inducible neural-specific protein-3 (BRINP3) as overexpressed in tumors, compared with normals. BRINP3 is a novel, normally brain restricted protein of unknown function. BRINP3 mRNA was expressed selectively in gonadotropinomas. Subcellular localization studies showed that BRINP3 was targeted to the mitochondria, but BRINP3 overexpression was unable to protect pituitary cells against programmed cell death induced by growth factor withdrawal. However, BRINP3 overexpression in pituitary gonadotrope cells promoted proliferation, migration, and invasion. A BRINP3 antibody was raised that demonstrated clustered expression of BRINP3 protein in gonadotropinomas and not in normal human pituitary samples. Thus, BRINP3 is a mitochondrially localized protein that is selectively up-regulated in human gonadotropinomas. Its actions to increase proliferation, migration, and invasion suggest it may play an important role in pituitary tumorigenesis. PMID- 17138657 TI - Dynamic changes in the cervical epithelial tight junction complex and differentiation occur during cervical ripening and parturition. AB - Cervical epithelia have numerous functions that include proliferation, differentiation, maintenance of fluid balance, protection from environmental hazards, and paracellular transport of solutes via tight junctions (TJs). Epithelial functions must be tightly regulated during pregnancy and parturition as the cervix undergoes extensive growth and remodeling. This study evaluated TJ proteins, as well as markers of epithelial cell differentiation in normal and cervical ripening defective mice to gain insights into how the permeability barrier is regulated during pregnancy and parturition. Although numerous TJ proteins are expressed in the nonpregnant cervix, claudins 1 and 2 are temporally regulated in pregnancy. Claudin 1 mRNA expression is increased, whereas claudin 2 expression declines. The cellular localization of claudin 1 shifts at the end of pregnancy (gestation d 18.75) to the plasma membrane in a lattice pattern, consistent with TJs in the apical cells. The timing of claudin 1-enriched TJs coincides with initiation of terminal differentiation of cervical squamous epithelia as evidenced by the increased expression of genes by differentiated epithelia late on gestation d 18. The cervical ripening defective steroid 5alpha reductase type 1 deficient mouse, which has an elevated local progesterone concentration, also has aberrant claudin 1 and 2 expressions, fails to form claudin 1-enriched TJs, and lacks normal expression of genes involved in epithelial terminal differentiation. These data suggest that changes in permeability barrier properties during cervical ripening are, in part, negatively regulated by progesterone, and that dynamic changes in barrier properties of the cervix occur during pregnancy and parturition. PMID- 17138658 TI - Undernutrition in utero augments systolic blood pressure and cardiac remodeling in adult mouse offspring: possible involvement of local cardiac angiotensin system in developmental origins of cardiovascular disease. AB - Evidence has emerged that undernutrition in utero is a risk factor for cardiovascular disorders in adulthood, along with genetic and environmental factors. Recently, the local expression of angiotensinogen and related bioactive substances has been demonstrated to play a pivotal role in cardiac remodeling, i.e. fibrosis and hypertrophy. The aim of the present study was to clarify the possible involvement of the local cardiac angiotensin system in fetal undernutrition-induced cardiovascular disorders. We developed a mouse model of undernutrition in utero by maternal food restriction, in which offspring (UN offspring) showed an increase in systolic blood pressure (8 wk of age, P < 0.05; and 16 wk, P < 0.01), perivascular fibrosis of the coronary artery (16 wk, P < 0.05) and cardiac cardiomegaly (16 wk, P < 0.01), and cardiomyocyte enlargement, concomitant with a significant augmentation of angiotensinogen (P < 0.05) and endothelin-1 (P < 0.01) mRNA expression and a tendency to increase in immunostaining for both angiotensin II and endothelin-1 in the left ventricles (16 wk). These findings suggest that fetal undernutrition activated the local cardiac angiotensin system-associated bioactive substances, which contributed, at least partly, to the development of cardiac remodeling in later life, in concert with the effects of increase in blood pressure. PMID- 17138659 TI - Insulin-like growth factor (IGF)-I controls prostate fibromuscular development: IGF-I inhibition prevents both fibromuscular and glandular development in eugonadal mice. AB - Although antiandrogen therapy has been shown effective in treating prostatic tumors, it is relatively ineffective in treating benign prostatic hyperplasia (BPH). In an attempt to understand better the role of androgens in the development of the normal prostate and BPH, we studied the relative effects of testosterone and IGF-I on the development of the two compartments of the prostate in castrated IGF-I((-/-)) male mice. Here we report that IGF-I stimulated the development of the fibromuscular compartment, but testosterone inhibited it (stromal epithelial ratio 2.17 vs. 0.83, respectively; P < 0.001). Testosterone also impaired IGF-I induced insulin receptor substrate-1 phosphorylation and cell division, and increased apoptosis in fibromuscular tissue. In sharp contrast IGF I and testosterone both stimulated the development of the glandular compartment individually and together. The combined effects were either additive or synergistic on compartment size, cell division, insulin receptor substrate-1 phosphorylation, and probasin production. Together they also had a greater inhibitory effect on apoptosis in gland tissue. To determine whether IGF-I inhibition would inhibit both fibromuscular and glandular compartments, we tested the effect of IGF binding protein-1 on prostate development in two different models: castrated Ames dwarf mice and eugonadal normal male mice. IGF binding protein-1 blocked bovine GH-induced fibromuscular and glandular development in both. It also inhibited epithelial cell division and increased apoptosis in both prostate compartments in the eugonadal mice. The observed discordance between IGF I and testosterone control of prostate compartment development might explain the relative failure of 5alpha-reductase inhibition in BPH and why testosterone inhibition might theoretically reduce gland volume but increase fibromuscular tissue. The work also provides a rationale for considering IGF-I inhibition as therapy for BPH to reduce the size of both prostate compartments. PMID- 17138661 TI - Gene replacement reveals a specific role for E-cadherin in the formation of a functional trophectoderm. AB - During mammalian embryogenesis the trophectoderm represents the first epithelial structure formed. The cell adhesion molecule E-cadherin is ultimately necessary for the transition from compacted morula to the formation of the blastocyst to ensure correct establishment of adhesion junctions in the trophectoderm. Here, we analyzed to what extent E-cadherin confers unique adhesion and signaling properties in trophectoderm formation in vivo. Using a gene replacement approach, we introduced N-cadherin cDNA into the E-cadherin genomic locus. We show that the expression of N-cadherin driven from the E-cadherin locus reflects the expression pattern of endogenous E-cadherin. Heterozygous mice co-expressing E- and N cadherin are vital and show normal embryonic development. Interestingly, N cadherin homozygous mutant embryos phenocopy E-cadherin-null mutant embryos. Upon removal of the maternal E-cadherin, we demonstrate that N-cadherin is able to provide sufficient cellular adhesion to mediate morula compaction, but is insufficient for the subsequent formation of a fully polarized functional trophectoderm. When ES cells were isolated from N-cadherin homozygous mutant embryos and teratomas were produced, these ES cells differentiated into a large variety of tissue-like structures. Importantly, different epithelial-like structures expressing N-cadherin were formed, including respiratory epithelia, squamous epithelia with signs of keratinization and secretory epithelia with goblet cells. Thus, N-cadherin can maintain epithelia in differentiating ES cells, but not during the formation of the trophectoderm. Our results point to a specific and unique function for E-cadherin during mouse preimplantation development. PMID- 17138662 TI - Ptpmeg is required for the proper establishment and maintenance of axon projections in the central brain of Drosophila. AB - Ptpmeg is a cytoplasmic tyrosine phosphatase containing FERM and PDZ domains. Drosophila Ptpmeg and its vertebrate homologs PTPN3 and PTPN4 are expressed in the nervous system, but their developmental functions have been unknown. We found that ptpmeg is involved in neuronal circuit formation in the Drosophila central brain, regulating both the establishment and the stabilization of axonal projection patterns. In ptpmeg mutants, mushroom body (MB) axon branches are elaborated normally, but the projection patterns in many hemispheres become progressively abnormal as the animals reach adulthood. The two branches of MB alpha/beta neurons are affected by ptpmeg in different ways; ptpmeg activity inhibits alpha lobe branch retraction while preventing beta lobe branch overextension. The phosphatase activity of Ptpmeg is essential for both alpha and beta lobe formation, but the FERM domain is required only for preventing alpha lobe retraction, suggesting that Ptpmeg has distinct roles in regulating the formation of alpha and beta lobes. ptpmeg is also important for the formation of the ellipsoid body (EB), where it influences the pathfinding of EB axons. ptpmeg function in neurons is sufficient to support normal wiring of both the EB and MB. However, ptpmeg does not act in either MB or EB neurons, implicating ptpmeg in the regulation of cell-cell signaling events that control the behavior of these axons. PMID- 17138663 TI - A Zn-finger/FH2-domain containing protein, FOZI-1, acts redundantly with CeMyoD to specify striated body wall muscle fates in the Caenorhabditis elegans postembryonic mesoderm. AB - Striated muscle development in vertebrates requires the redundant functions of multiple members of the MyoD family. Invertebrates such as Drosophila and Caenorhabditis elegans contain only one MyoD homolog in each organism. Earlier observations suggest that factors outside of the MyoD family might function redundantly with MyoD in striated muscle fate specification in these organisms. However, the identity of these factors has remained elusive. Here, we describe the identification and characterization of FOZI-1, a putative transcription factor that functions redundantly with CeMyoD (HLH-1) in striated body wall muscle (BWM) fate specification in the C. elegans postembryonic mesoderm. fozi-1 encodes a novel nuclear-localized protein with motifs characteristic of both transcription factors and actin-binding proteins. We show that FOZI-1 shares the same expression pattern as CeMyoD in the postembryonic mesodermal lineage, the M lineage, and that fozi-1-null mutants exhibit similar M lineage-null defects to those found in animals lacking CeMyoD in the M lineage (e.g. loss of a fraction of M lineage-derived BWMs). Interestingly, fozi-1-null mutants with a reduced level of CeMyoD lack most, if not all, M lineage-derived BWMs. Our results indicate that FOZI-1 and the Hox factor MAB-5 function redundantly with CeMyoD in the specification of the striated BWM fate in the C. elegans postembryonic mesoderm, implicating a remarkable level of complexity for the production of a simple striated musculature in C. elegans. PMID- 17138664 TI - The TGFbeta intracellular effector Smad3 regulates neuronal differentiation and cell fate specification in the developing spinal cord. AB - Here we show that Smad3, a transforming growth factor beta (TGFbeta)/activin signaling effector, is expressed in discrete progenitor domains along the dorsoventral axis of the developing chick spinal cord. Restriction of Smad3 expression to the dP6-p2 and p3 domains together with exclusion from the motoneuron progenitor domain, are the result of the activity of key transcription factors responsible for patterning the neural tube. Smad3-mediated TGFbeta activity promotes cell-cycle exit and neurogenesis by inhibiting the expression of Id proteins, and activating the expression of neurogenic factors and the cyclin-dependent-kinase-inhibitor p27(kip1). Furthermore, Smad3 activity induces differentiation of selected neuronal subtypes at the expense of other subtypes. Within the intermediate and ventral domains, Smad3 promotes differentiation of ventral interneurons at the expense of motoneuron generation. Consequently, the absence of Smad3 expression from the motoneuron progenitor domain during pattern formation of the neural tube is a prerequisite for the correct generation of spinal motoneurons. PMID- 17138665 TI - The making of Wnt: new insights into Wnt maturation, sorting and secretion. PMID- 17138666 TI - Functional redundancy among Nanos proteins and a distinct role of Nanos2 during male germ cell development. AB - The mouse Nanos proteins, Nanos2 and Nanos3, are required for germ cell development and share a highly conserved zinc-finger domain. The expression patterns of these factors during development, however, differ from each other. Nanos3 expression in the mouse embryo commences in the primordial germ cells (PGCs) just after their formation, and a loss of this protein results in the germ cell-less phenotype in both sexes. By contrast, Nanos2 expression begins only in male PGCs after their entry into the genital ridge and a loss of this protein results in a male germ cell deficiency, irrespective of the co-expression of Nanos3 in these cells. These results indicate that these two Nanos proteins have distinct functions, which depend on the time and place of their expression. To further elucidate this, we have generated transgenic mouse lines that express Nanos2 under the control of the Oct4DeltaPE promoter and examined Nanos2 function in a Nanos3-null genetic background. We find that ectopically produced Nanos2 protein rescues the Nanos3-null defects, because the germ cells fully develop in both sexes in the transgenic mice. This result indicates that Nanos2 can substitute for Nanos3 during early PGC development. By contrast, our current data show that Nanos3 does not rescue the defects in Nanos2-null mice. Our present findings thus indicate that there are redundant functions of the Nanos proteins in early PGC development, but that Nanos2 has a distinct function during male germ cell development in the mouse. PMID- 17138667 TI - Tbx5 is dispensable for forelimb outgrowth. AB - Tbx5 is essential for initiation of the forelimb, and its deletion in mice results in the failure of forelimb formation. Misexpression of dominant-negative forms of Tbx5 results in limb truncations, suggesting Tbx5 is also required for forelimb outgrowth. Here we show that Tbx5 is expressed throughout the limb mesenchyme in progenitors of cartilage, tendon and muscle. Using a tamoxifeninducible Cre transgenic line, we map the time frame during which Tbx5 is required for limb development. We show that deletion of Tbx5 subsequent to limb initiation does not impair limb outgrowth. Furthermore, we distinguish two distinct phases of limb development: a Tbx5-dependent limb initiation phase, followed by a Tbx5-independent limb outgrowth phase. In humans, mutations in the T-box transcription factor TBX5 are associated with the dominant disorder Holt Oram syndrome (HOS), which is characterised by malformations in the forelimb and heart. Our results demonstrate a short temporal requirement for Tbx5 during early limb development, and suggest that the defects found in HOS arise as a result of disrupted TBX5 function during this narrow time window. PMID- 17138668 TI - Using the principle of entropy maximization to infer genetic interaction networks from gene expression patterns. AB - We describe a method based on the principle of entropy maximization to identify the gene interaction network with the highest probability of giving rise to experimentally observed transcript profiles. In its simplest form, the method yields the pairwise gene interaction network, but it can also be extended to deduce higher-order interactions. Analysis of microarray data from genes in Saccharomyces cerevisiae chemostat cultures exhibiting energy metabolic oscillations identifies a gene interaction network that reflects the intracellular communication pathways that adjust cellular metabolic activity and cell division to the limiting nutrient conditions that trigger metabolic oscillations. The success of the present approach in extracting meaningful genetic connections suggests that the maximum entropy principle is a useful concept for understanding living systems, as it is for other complex, nonequilibrium systems. PMID- 17138669 TI - Production of hydrogen peroxide in the atmosphere of a Snowball Earth and the origin of oxygenic photosynthesis. AB - During Proterozoic time, Earth experienced two intervals with one or more episodes of low-latitude glaciation, which are probable "Snowball Earth" events. Although the severity of the historical glaciations is debated, theoretical "hard Snowball" conditions are associated with the nearly complete shutdown of the hydrological cycle. We show here that, during such long and severe glacial intervals, a weak hydrological cycle coupled with photochemical reactions involving water vapor would give rise to the sustained production of hydrogen peroxide. The photochemical production of hydrogen peroxide has been proposed previously as the primary mechanism for oxidizing the surface of Mars. During a Snowball, hydrogen peroxide could be stored in the ice; it would then be released directly into the ocean and the atmosphere upon melting and could mediate global oxidation events in the aftermath of the Snowball, such as that recorded in the Fe and Mn oxides of the Kalahari Manganese Field, deposited after the Paleoproterozoic low-latitude Makganyene glaciation. Low levels of peroxides and molecular oxygen generated during Archean and earliest Proterozoic non-Snowball glacial intervals could have driven the evolution of oxygen-mediating and -using enzymes and thereby paved the way for the eventual appearance of oxygenic photosynthesis. PMID- 17138670 TI - GABA and Gi/o differentially control circadian rhythms and synchrony in clock neurons. AB - Neurons in the mammalian suprachiasmatic nuclei (SCN) generate daily rhythms in physiology and behavior, but it is unclear how they maintain and synchronize these rhythms in vivo. We hypothesized that parallel signaling pathways in the SCN are required to synchronize rhythms in these neurons for coherent output. We recorded firing and clock-gene expression patterns while blocking candidate signaling pathways for at least 8 days. GABA(A) and GABA(B) antagonism increased circadian peak firing rates and rhythm precision of cultured SCN neurons, but G(i/o) did not impair synchrony or rhythmicity. In contrast, inhibiting G(i/o) with pertussis toxin abolished rhythms in most neurons and desynchronized the population, phenocopying the loss of vasoactive intestinal polypeptide (VIP). Daily VIP receptor agonist treatment restored synchrony and rhythmicity to VIP(-/ ) SCN cultures during continuous GABA receptor antagonism but not during G(i/o) blockade. Pertussis toxin did not affect circadian cycling of the liver, suggesting that G(i/o) plays a specialized role in maintaining SCN rhythmicity. We conclude that endogenous GABA controls the amplitude of SCN neuronal rhythms by reducing daytime firing, whereas G(i/o) signaling suppresses nighttime firing, and it is necessary for synchrony among SCN neurons. We propose that G(i/o), not GABA activity, converges with VIP signaling to maintain and coordinate rhythms among SCN neurons. PMID- 17138671 TI - Quantitative proteomic analysis of distinct mammalian Mediator complexes using normalized spectral abundance factors. AB - Components of multiprotein complexes are routinely determined by using proteomic approaches. However, this information lacks functional content except when new complex members are identified. To analyze quantitatively the abundance of proteins in human Mediator we used normalized spectral abundance factors generated from shotgun proteomics data sets. With this approach we define a common core of mammalian Mediator subunits shared by alternative forms that variably associate with the kinase module and RNA polymerase (pol) II. Although each version of affinity-purified Mediator contained some kinase module and RNA pol II, Mediator purified through F-Med26 contained the most RNA pol II and the least kinase module as demonstrated by the normalized spectral abundance factor approach. The distinct forms of Mediator were functionally characterized by using a transcriptional activity assay, where F-Med26 Mediator/RNA pol II was the most active. This method of protein complex visualization has important implications for the analysis of multiprotein complexes and assembly of protein interaction networks. PMID- 17138672 TI - Viral protease cleavage of inhibitor of kappaBalpha triggers host cell apoptosis. AB - Apoptosis is an innate immune response to viral infection that limits viral replication. However, the mechanisms by which cells detect viral infection and activate apoptosis are not completely understood. We now show that during Coxsackievirus infection, the viral protease 3C(pro) cleaves inhibitor of kappaBalpha (IkappaBalpha). A proteolytic fragment of IkappaBalpha then forms a stable complex with NF-kappaB, translocates to the nucleus, and inhibits NF kappaB transactivation, increasing apoptosis and decreasing viral replication. In contrast, cells with reduced IkappaBalpha expression are more susceptible to viral infection, with less apoptosis and more viral replication. IkappaBalpha thus acts as a sensor of viral infection. Cleavage of host proteins by pathogen proteases is a novel mechanism by which the host recognizes and responds to viral infection. PMID- 17138673 TI - Evidence-based medicines for children: Important implications for new therapies at all ages. PMID- 17138674 TI - Long-term treprostinil in pulmonary arterial hypertension: Is the glass half full or half empty? PMID- 17138675 TI - The mycobacterial mystery. PMID- 17138676 TI - New ways in respiratory genetics. PMID- 17138677 TI - Genotype-corrected reference values for serum angiotensin-converting enzyme. AB - The deletion (D)/insertion (I) polymorphism in intron 16 of the angiotensin converting enzyme (ACE) gene has the greatest impact on serum ACE level in Caucasians of any factor yet discovered. The aim of the present study was to establish new ACE genotype-corrected normal ranges for serum ACE level in a population of central European origin. After a medical examination, 159 healthy Caucasians volunteered to donate blood for the study. ACE genotypes were assessed by PCR and serum ACE levels were determined using two different kinetic tests. The distribution of the D/I polymorphism of the ACE gene was in accordance with the Hardy-Weinberg equilibrium. Serum ACE levels and ACE genotypes correlated significantly, with the highest serum ACE levels in subjects with ACE genotype D/D, and the lowest serum ACE levels in subjects with genotype I/I (mean+/-sd, assay 1: D/D 59.3+/-15.1 U x L(-1), D/I 45.5+/-15.2 U x L(-1), I/I 34.8+/-13.7 U x L(-1); assay 2: D/D 43.7+/-14.1 U x L(-1), D/I 33.7+/-12.1 U x L(-1), I/I 25.4+/-9.5 U x L(-1)). Although they gave different absolute values of serum ACE levels, the results of the two test kits correlated significantly. In conclusion, the present authors recommend the use of new, genotype-specific reference values for serum angiotensin-converting enzyme levels, especially to improve the sensitivity and specificity of tests for angiotensin-converting enzyme in the follow-up of sarcoidosis. PMID- 17138678 TI - Respiratory infections associated with nontuberculous mycobacteria in non-HIV patients. AB - The incidence of nontuberculous mycobacteria (NTM) pulmonary diseases in HIV negative patients was studied prospectively from January 1, 2001 to December 31, 2003 by 32 sentinel sites distributed throughout France. In total, 262 patients who yielded NTM isolates from respiratory clinical specimens, met the bacteriological, radiological and clinical criteria established by the American Thoracic Society for NTM respiratory disease. Among the 262 NTM isolates, 234 were slow-growing mycobacteria (125 Mycobacterium avium-intracellulare complex (MAC), 66 M. xenopi, 34 M. kansasii) and 28 were rapidly growing mycobacteria (25 M. abscessus complex). In the Paris area, M. xenopi was the most frequently isolated species, followed by MAC. Most patients (>50%), except those with M. kansasii, had underlying predisposing factors such as pre-existing pulmonary disease or immune deficiency. Asthenia, weight loss, chronic cough and dyspnoea were the most common clinical symptoms. The classical radiological appearance of NTM infections was indistinguishable from that observed in patients with pulmonary tuberculosis. In summary, the incidence of nontuberculous mycobacteria pulmonary infections in HIV-negative patients was estimated at 0.74, 0.73 and 0.72 cases per 100,000 inhabitants in 2001, 2002 and 2003, respectively. PMID- 17138679 TI - Mortality in COPD: Role of comorbidities. AB - Chronic obstructive pulmonary disease (COPD) represents an increasing burden throughout the world. COPD-related mortality is probably underestimated because of the difficulties associated with identifying the precise cause of death. Respiratory failure is considered the major cause of death in advanced COPD. Comorbidities such as cardiovascular disease and lung cancer are also major causes and, in mild-to-moderate COPD, are the leading causes of mortality. The links between COPD and these conditions are not fully understood. However, a link through the inflammation pathway has been suggested, as persistent low-grade pulmonary and systemic inflammation, both known risk factors for cardiovascular disease and cancer, are present in COPD independent of cigarette smoking. Lung specific measurements, such as forced expiratory volume in one second (FEV(1)), predict mortality in COPD and in the general population. However, composite tools, such as health-status measurements (e.g. St George's Respiratory Questionnaire) and the BODE index, which incorporates Body mass index, lung function (airflow Obstruction), Dyspnoea and Exercise capacity, predict mortality better than FEV(1) alone. These multidimensional tools may be more valuable because, unlike predictive approaches based on single parameters, they can reflect the range of comorbidities and the complexity of underlying mechanisms associated with COPD. The current paper reviews the role of comorbidities in chronic obstructive pulmonary disease mortality, the putative underlying pathogenic link between chronic obstructive pulmonary disease and comorbid conditions (i.e. inflammation), and the tools used to predict chronic obstructive pulmonary disease mortality. PMID- 17138680 TI - Evolution and respiratory genetics. AB - Evolution is a plausible explanation for between-population differences in particular allele frequencies if: the genes involved have related functions; the heterogeneous alleles involved have similar functional consequences; the involved genes are not linked chromosomally; and the patterns observed would result in a biologically plausible, survival-enhancing gene-environment interaction. However, possible evolutionary effects have to be differentiated from founder effects and random genetic drift. The current authors have noted the existence of a consistent pattern of allelic frequencies in genes related to T-helper 2 (Th2) immune responses in humans of different ancestral backgrounds, residing in climatically similar regions. Th2 responses are thought to have evolved in mammals to resist infection by parasites, particularly helminths. Modern man arose in tropical Africa where helminths thrived. Relatively recently, humans migrated to cooler or drier climates where most helminths struggled to reproduce. The genetic tendency to strong Th2 responses may have become a health liability, the reduction in risk from parasites being counterbalanced by an increased inherited propensity to atopic or allergic diseases. The pattern noted by the present authors includes specific alleles of interleukin-4 and its receptor, interleukin-13, interleukin-10, the beta chain of the high-affinity receptor for immunoglobulin E, the beta(1)-adrenergic receptor, and the alpha chain of tumour necrosis factor. These population-specific polymorphism profiles are likely to be relevant in current disease patterns. The high incidence of asthma in migrants from tropical locations to affluent temperate countries is likely to be related to these patterns. Of even more concern is the possibility that increasing westernisation among the approximately 2 billion people living in the tropics will produce rapidly increasing levels of asthma, as these populations have a high genetic predisposition to allergic disease. PMID- 17138681 TI - Transbronchial and transoesophageal (ultrasound-guided) needle aspirations for the analysis of mediastinal lesions. AB - A tissue diagnosis of mediastinal nodes is frequently needed for accurate lung cancer staging as well as the assessment of mediastinal masses. Transbronchial needle aspiration (TBNA) is a safe procedure that is performed during routine bronchoscopy. Provided mediastinal metastases are confirmed, TBNA has a high impact on patient management. Unfortunately, TBNA remains underused in current daily practice, mainly due to the lack of real-time needle visualisation. The introduction of echo-endoscopes has overcome this problem. Endobronchial ultrasound-guided TBNA (EBUS-TBNA) allows real-time controlled tissue sampling of paratracheal, subcarinal and hilar lymph nodes. Mediastinal lymph nodes located adjacent to the oesophagus can be assessed by transoesophageal ultrasound-guided fine needle aspiration (EUS-FNA). Owing to the complementary reach of EBUS-TBNA and EUS-FNA in assessing different regions of the mediastinum, recent studies suggest that complete and accurate mediastinal staging can be achieved by the combination of both procedures. It is expected that implementation of minimally invasive endoscopic methods of endobronchial ultrasound-guided transbronchial needle aspiration and transoesophageal ultrasound-guided fine needle aspiration will reduce the need for surgical staging of lung cancer significantly. PMID- 17138682 TI - Severe airflow obstruction and eosinophilic lung disease after Stevens-Johnson syndrome. AB - Respiratory involvement is a frequent complication of Stevens-Johnson syndrome (SJS). However, there are very few convincing reports of persistent pulmonary sequelae, as demonstrated by spirometry, radiology and pathology. The current study presents a case of a 13-yr-old female with T-cell acute lymphocytic leukaemia who developed persistent, severe, obstructive lung disease following an episode of SJS. A lung biopsy demonstrated bronchiolar submucosal fibrosis consistent with constrictive bronchiolitis, as well as eosinophilic micro abscesses, which, to the current authors' knowledge, has not been previously described. The present study illustrates specific histopathological features that highlight a possible association between Stevens-Johnson syndrome, constrictive bronchiolitis and eosinophilic micro-abscesses. The eosinophils may be associated with permanent mucosal damage, as seen in the present case, by releasing mediators that have a pro-fibrogenetic role. However, further investigation is warranted. PMID- 17138683 TI - Caucasian race and lung function: Time to revisit the racial groups used in reference values. PMID- 17138684 TI - Three new cases of apparent occupational asthma in swine confinement facility employees. PMID- 17138685 TI - Components of airway disease. PMID- 17138686 TI - The specificity of interferon-gamma-based blood tests in the identification of latent tuberculosis infection. PMID- 17138687 TI - Lymphangioleiomyomatosis. PMID- 17138688 TI - Cost-optimisation of screening for latent tuberculosis in close contacts. PMID- 17138689 TI - Therapy of exophytic bronchial tumorous stenosis by flexible cryoprobe. PMID- 17138690 TI - Is forced expiratory volume in six seconds a valid alternative to forced vital capacity? PMID- 17138691 TI - Epidemiology and costs of COPD. PMID- 17138692 TI - Cryptogenic organising pneumonia. PMID- 17138693 TI - C-23 hydroxylation by Arabidopsis CYP90C1 and CYP90D1 reveals a novel shortcut in brassinosteroid biosynthesis. AB - Brassinosteroids (BRs) are biosynthesized from campesterol via several cytochrome P450 (P450)-catalyzed oxidative reactions. We report the functional characterization of two BR-biosynthetic P450s from Arabidopsis thaliana: CYP90C1/ROTUNDIFOLIA3 and CYP90D1. The cyp90c1 cyp90d1 double mutant exhibits the characteristic BR-deficient dwarf phenotype, although the individual mutants do not display this phenotype. These data suggest redundant roles for these P450s. In vitro biochemical assays using insect cell-expressed proteins revealed that both CYP90C1 and CYP90D1 catalyze C-23 hydroxylation of various 22-hydroxylated BRs with markedly different catalytic efficiencies. Both enzymes preferentially convert 3-epi-6-deoxocathasterone, (22S,24R)-22-hydroxy-5alpha-ergostan-3-one, and (22S,24R)-22-hydroxyergost-4-en-3-one to 23-hydroxylated products, whereas they are less active on 6-deoxocathasterone. Likewise, cyp90c1 cyp90d1 plants were deficient in 23-hydroxylated BRs, and in feeding experiments using exogenously supplied intermediates, only 23-hydroxylated BRs rescued the growth deficiency of the cyp90c1 cyp90d1 mutant. Thus, CYP90C1 and CYP90D1 are redundant BR C-23 hydroxylases. Moreover, their preferential substrates are present in the endogenous Arabidopsis BR pool. Based on these results, we propose C-23 hydroxylation shortcuts that bypass campestanol, 6-deoxocathasterone, and 6 deoxoteasterone and lead directly from (22S,24R)-22-hydroxy-5alpha-ergostan-3-one and 3-epi-6-deoxocathasterone to 3-dehydro-6-deoxoteasterone and 6 deoxotyphasterol. PMID- 17138694 TI - SUPPRESSOR OF FRIGIDA4, encoding a C2H2-Type zinc finger protein, represses flowering by transcriptional activation of Arabidopsis FLOWERING LOCUS C. AB - FLOWERING LOCUS C (FLC), a strong floral repressor, is one of the central regulators of flowering in Arabidopsis thaliana. The expression of FLC is increased by FRIGIDA (FRI) but decreased by vernalization, a long period of cold exposure that accelerates flowering. Although many aspects of FLC regulation have been reported, it is not known how FLC is transcriptionally activated by FRI at the molecular level. We isolated suppressor of FRIGIDA4 (suf4), a mutant that flowers early as a result of low FLC expression. SUF4 encodes a nuclear-localized protein with two C2H2-type zinc finger motifs and a Pro-rich domain. SUF4 protein interacts with FRI and FRIGIDA-LIKE1 (FRL1), two genes for which single mutations have the same phenotype as suf4. SUF4 also bound to the promoter of FLC in a chromatin immunoprecipitation assay, suggesting that SUF4 acts as a transcriptional activator of FLC after forming a complex with FRI and FRL1. In addition, suf4 suppresses luminidependens (ld), a late-flowering mutation that causes an increase of FLC, and SUF4 protein directly interacts with LD. Thus, we propose that LD binds to SUF4 to suppress its activity in the absence of FRI. PMID- 17138695 TI - The rice tapetum degeneration retardation gene is required for tapetum degradation and anther development. AB - In flowering plants, tapetum degeneration is proposed to be triggered by a programmed cell death (PCD) process during late stages of pollen development; the PCD is thought to provide cellular contents supporting pollen wall formation and to allow the subsequent pollen release. However, the molecular basis regulating tapetum PCD in plants remains poorly understood. We report the isolation and characterization of a rice (Oryza sativa) male sterile mutant tapetum degeneration retardation (tdr), which exhibits degeneration retardation of the tapetum and middle layer as well as collapse of microspores. The TDR gene is preferentially expressed in the tapetum and encodes a putative basic helix-loop helix protein, which is likely localized to the nucleus. More importantly, two genes, Os CP1 and Os c6, encoding a Cys protease and a protease inhibitor, respectively, were shown to be the likely direct targets of TDR through chromatin immunoprecipitation analyses and the electrophoretic mobility shift assay. These results indicate that TDR is a key component of the molecular network regulating rice tapetum development and degeneration. PMID- 17138696 TI - A ferroxidation/permeation iron uptake system is required for virulence in Ustilago maydis. AB - In the smut fungus Ustilago maydis, a tightly regulated cAMP signaling cascade is necessary for pathogenic development. Transcriptome analysis using whole genome microarrays set up to identify putative target genes of the protein kinase A catalytic subunit Adr1 revealed nine genes with putative functions in two high affinity iron uptake systems. These genes locate to three gene clusters on different chromosomes and include the previously identified complementing siderophore auxotroph genes sid1 and sid2 involved in siderophore biosynthesis. Transcription of all nine genes plus three additional genes associated with the gene clusters was also coregulated by iron through the Urbs1 transcription factor. Two components of a high-affinity iron uptake system were characterized in more detail: fer2, encoding a high-affinity iron permease; and fer1, encoding an iron multicopper oxidase. Fer2 localized to the plasma membrane and complemented an ftr1 mutant of Saccharomyces cerevisiae lacking a high-affinity iron permease. During pathogenic development, fer2 expression was confined to the phase of hyphal proliferation inside the plant. fer2 as well as fer1 deletion mutants were strongly affected in virulence. These data highlight the importance of the high-affinity iron uptake system via an iron permease and a multicopper oxidase for biotrophic development in the U. maydis/maize (Zea mays) pathosystem. PMID- 17138697 TI - CONSTANS and the CCAAT box binding complex share a functionally important domain and interact to regulate flowering of Arabidopsis. AB - The CCT (for CONSTANS, CONSTANS-LIKE, TOC1) domain is found in 45 Arabidopsis thaliana proteins involved in processes such as photoperiodic flowering, light signaling, and regulation of circadian rhythms. We show that this domain exhibits similarities to yeast HEME ACTIVATOR PROTEIN2 (HAP2), which is a subunit of the HAP2/HAP3/HAP5 trimeric complex that binds to CCAAT boxes in eukaryotic promoters. Moreover, we demonstrate that CONSTANS (CO), which promotes Arabidopsis flowering, interacts with At HAP3 and At HAP5 in yeast, in vitro, and in planta. Mutations in CO that delay flowering affect residues highly conserved between CCT and the DNA binding domain of HAP2. Taken together, these data suggest that CO might replace At HAP2 in the HAP complex to form a trimeric CO/At HAP3/At HAP5 complex. Flowering was delayed by overexpression of At HAP2 or At HAP3 throughout the plant or in phloem companion cells, where CO is expressed. This phenotype was correlated with reduced abundance of FLOWERING LOCUS T (FT) mRNA and no change in CO mRNA levels. At HAP2 or At HAP3 overexpression may therefore impair formation of a CO/At HAP3/At HAP5 complex leading to reduced expression of FT. During plant evolution, the number of genes encoding HAP proteins was greatly amplified, and these proteins may have acquired novel functions, such as mediating the effect of CCT domain proteins on gene expression. PMID- 17138698 TI - Two cytosolic glutamine synthetase isoforms of maize are specifically involved in the control of grain production. AB - The roles of two cytosolic maize glutamine synthetase isoenzymes (GS1), products of the Gln1-3 and Gln1-4 genes, were investigated by examining the impact of knockout mutations on kernel yield. In the gln1-3 and gln1-4 single mutants and the gln1-3 gln1-4 double mutant, GS mRNA expression was impaired, resulting in reduced GS1 protein and activity. The gln1-4 phenotype displayed reduced kernel size and gln1-3 reduced kernel number, with both phenotypes displayed in gln1-3 gln1-4. However, at maturity, shoot biomass production was not modified in either the single mutants or double mutants, suggesting a specific impact on grain production in both mutants. Asn increased in the leaves of the mutants during grain filling, indicating that it probably accumulates to circumvent ammonium buildup resulting from lower GS1 activity. Phloem sap analysis revealed that unlike Gln, Asn is not efficiently transported to developing kernels, apparently causing reduced kernel production. When Gln1-3 was overexpressed constitutively in leaves, kernel number increased by 30%, providing further evidence that GS1-3 plays a major role in kernel yield. Cytoimmunochemistry and in situ hybridization revealed that GS1-3 is present in mesophyll cells, whereas GS1-4 is specifically localized in the bundle sheath cells. The two GS1 isoenzymes play nonredundant roles with respect to their tissue-specific localization. PMID- 17138699 TI - Wax-deficient anther1 is involved in cuticle and wax production in rice anther walls and is required for pollen development. AB - In vegetative leaf tissues, cuticles including cuticular waxes are important for protection against nonstomatal water loss and pathogen infection as well as for adaptations to environmental stress. However, their roles in the anther wall are rarely studied. The innermost layer of the anther wall (the tapetum) is essential for generating male gametes. Here, we report the characterization of a T-DNA insertional mutant in the Wax-deficient anther1 (Wda1) gene of rice (Oryza sativa), which shows significant defects in the biosynthesis of very-long-chain fatty acids in both layers. This gene is strongly expressed in the epidermal cells of anthers. Scanning electron microscopy analyses showed that epicuticular wax crystals were absent in the outer layer of the anther and that microspore development was severely retarded and finally disrupted as a result of defective pollen exine formation in the mutant anthers. These biochemical and developmental defects in tapetum found in wda1 mutants are earlier events than those in other male-sterile mutants, which showed defects of lipidic molecules in exine. Our findings provide new insights into the biochemical and developmental aspects of the role of waxes in microspore exine development in the tapetum as well as the role of epicuticular waxes in anther expansion. PMID- 17138700 TI - The POLARIS peptide of Arabidopsis regulates auxin transport and root growth via effects on ethylene signaling. AB - The rate and plane of cell division and anisotropic cell growth are critical for plant development and are regulated by diverse mechanisms involving several hormone signaling pathways. Little is known about peptide signaling in plant growth; however, Arabidopsis thaliana POLARIS (PLS), encoding a 36-amino acid peptide, is required for correct root growth and vascular development. Mutational analysis implicates a role for the peptide in hormone responses, but the basis of PLS action is obscure. Using the Arabidopsis root as a model to study PLS action in plant development, we discovered a link between PLS, ethylene signaling, auxin homeostasis, and microtubule cytoskeleton dynamics. Mutation of PLS results in an enhanced ethylene-response phenotype, defective auxin transport and homeostasis, and altered microtubule sensitivity to inhibitors. These defects, along with the short-root phenotype, are suppressed by genetic and pharmacological inhibition of ethylene action. PLS expression is repressed by ethylene and induced by auxin. Our results suggest a mechanism whereby PLS negatively regulates ethylene responses to modulate cell division and expansion via downstream effects on microtubule cytoskeleton dynamics and auxin signaling, thereby influencing root growth and lateral root development. This mechanism involves a regulatory loop of auxin-ethylene interactions. PMID- 17138702 TI - Pharmacokinetics, metabolism, and oral bioavailability of the DNA methyltransferase inhibitor 5-fluoro-2'-deoxycytidine in mice. AB - PURPOSE: In vivo, 5-fluoro-2'-deoxycytidine (FdCyd) is rapidly and sequentially converted to 5-fluoro-2'-deoxyuridine, 5-fluorouracil, and 5-fluorouridine. The i.v. combination of FdCyd and 3,4,5,6-tetrahydrouridine (THU), a cytidine deaminase (CD) inhibitor that blocks the first metabolic step in FdCyd catabolism, is being investigated clinically for its ability to inhibit DNA methyltransferase. However, the full effects of THU on FdCyd metabolism and pharmacokinetics are unknown. We aimed to characterize the pharmacokinetics, metabolism, and bioavailability of FdCyd with and without THU in mice. EXPERIMENTAL DESIGN: We developed a sensitive high-performance liquid chromatography tandem mass spectrometry assay to quantitate FdCyd and metabolites in mouse plasma. Mice were dosed i.v. or p.o. with 25 mg/kg FdCyd with or without coadministration of 100 mg/kg THU p.o. or i.v. RESULTS: The oral bioavailability of FdCyd alone was approximately 4%. Coadministration with THU increased exposure to FdCyd and decreased exposure to its metabolites; i.v. and p.o. coadministration of THU increased exposure to p.o. FdCyd by 87- and 58-fold, respectively. FdCyd exposure after p.o. FdCyd with p.o. THU was as much as 54% that of i.v. FdCyd with i.v. THU. CONCLUSIONS: FdCyd is well absorbed but undergoes substantial first-pass catabolism by CD to potentially toxic metabolites that do not inhibit DNA methyltransferase. THU is sufficiently bioavailable to reduce the first-pass effect of CD on FdCyd. Oral coadministration of THU and FdCyd is a promising approach that warrants clinical testing because it may allow maintaining effective FdCyd concentrations on a chronic basis, which would be an advantage over other DNA methyltransferase inhibitors that are currently approved or in development. PMID- 17138701 TI - Plant N-glycan processing enzymes employ different targeting mechanisms for their spatial arrangement along the secretory pathway. AB - The processing of N-linked oligosaccharides in the secretory pathway requires the sequential action of a number of glycosidases and glycosyltransferases. We studied the spatial distribution of several type II membrane-bound enzymes from Glycine max, Arabidopsis thaliana, and Nicotiana tabacum. Glucosidase I (GCSI) localized to the endoplasmic reticulum (ER), alpha-1,2 mannosidase I (ManI) and N acetylglucosaminyltransferase I (GNTI) both targeted to the ER and Golgi, and beta-1,2 xylosyltransferase localized exclusively to Golgi stacks, corresponding to the order of expected function. ManI deletion constructs revealed that the ManI transmembrane domain (TMD) contains all necessary targeting information. Likewise, GNTI truncations showed that this could apply to other type II enzymes. A green fluorescent protein chimera with ManI TMD, lengthened by duplicating its last seven amino acids, localized exclusively to the Golgi and colocalized with a trans-Golgi marker (ST52-mRFP), suggesting roles for protein-lipid interactions in ManI targeting. However, the TMD lengths of other plant glycosylation enzymes indicate that this mechanism cannot apply to all enzymes in the pathway. In fact, removal of the first 11 amino acids of the GCSI cytoplasmic tail resulted in relocalization from the ER to the Golgi, suggesting a targeting mechanism relying on protein-protein interactions. We conclude that the localization of N-glycan processing enzymes corresponds to an assembly line in the early secretory pathway and depends on both TMD length and signals in the cytoplasmic tail. PMID- 17138703 TI - Did you take your medications? The dentist's role in helping patients adhere to their drug regimen. PMID- 17138704 TI - Musculoskeletal disorders. PMID- 17138705 TI - A 'commodity-based model'. PMID- 17138706 TI - Periodontal treatment does not reduce the risk of preterm delivery, study finds. PMID- 17138708 TI - Molecule that induces biofilm dispersion discovered. PMID- 17138709 TI - A meta-analysis of six-month studies of antiplaque and antigingivitis agents. AB - BACKGROUND AND METHODS: The author conducted a systematic review of the literature to evaluate the efficacy of antigingivitis and antiplaque products in six-month trials. He searched electronic databases for six-month randomized clinical studies that evaluated both antiplaque and antigingivitis properties of dentifrices or mouthrinses. In addition, the author solicited unpublished studies from manufacturers. RESULTS: Seventeen studies support the antiplaque, antigingivitis effects of dentifrices containing 0.30 percent triclosan, 2.0 percent Gantrez copolymer. There was no evidence of efficacy for triclosan products containing either soluble pyrophosphate or zinc citrate. Dentifrices with stannous fluoride had statistically significant, but marginally clinically significant, evidence of an antiplaque effect; however, there was both a statistically and clinically significant antigingivitis effect. The largest body of studies (21 studies) supported the efficacy of mouthrinses with essential oils. A smaller body of studies (seven) supported a strong antiplaque, antigingivitis effect of mouthrinses with 0.12 percent chlorhexidine. Results for mouthrinses with cetylpyridinium chloride varied and depended on the product's formula. CONCLUSIONS: The studies in this systematic review provide strong evidence of the antiplaque, antigingivitis effects of multiple agents. These results support the use of these agents as part of a typical oral hygiene regimen. PMID- 17138710 TI - Chemomechanical caries removal in children: efficacy and efficiency. AB - BACKGROUND: The authors investigated the effectiveness of chemomechanical caries removal (CMCR) compared with the traditional method (TM) of caries removal using a round bur when treating dentinal-depth occlusal lesions with minimal enamel access in primary molars. The authors also compare CMCR with TM to determine if it had a higher efficacy and could be used more frequently without the subject's having to undergo local anesthesia. METHODS: The authors collected data from 50 children during operative appointments at which caries was removed using one of the two methods. RESULTS: Complete caries removal within 15 minutes was achieved in only 57.7 percent of the CMCR-treated teeth. In 42.3 percent of these teeth, residual caries was removed using TM. CMCR was almost eight times more time consuming than was TM when used to excavate dentinal-depth occlusal lesions with minimal cavitation. There was no significant difference between CMCR and TM in the number of subjects who needed to undergo local anesthesia. CONCLUSIONS: The authors found no direct clinical advantage in using CMCR over using TM for treating occlusal dentinal lesions with minimal cavitation in pediatric patients. PMID- 17138711 TI - Metastatic tumors in the jaws: a retrospective study of 114 cases. AB - BACKGROUND: Malignancies involving the bones are metastatic tumors more commonly than primary tumors. In this retrospective study, the authors review metastatic disease in the jaws. METHODS: The authors retrieved cases of metastatic disease in the jaws over a 45-year period from the pathology archives at the University of Michigan School of Dentistry, Ann Arbor, and Indiana University School of Dentistry, Indianapolis. RESULTS: The authors conducted a retrospective analysis of 114 cases of metastatic disease in the jaws and found that approximately 60 percent of subjects had no history of malignancy. The sex distribution was equivalent. Mandibular predilection was more prominent in females than in males. Metastases from the breast were significantly greater than those from the lung and prostate (P < or = .05), the second and third most frequent sites, respectively. Women exhibited twice as many jaw metastases as did men 31 to 40 years of age and significantly fewer metastases than did men 71 to 80 years of age (P < or = .05). CONCLUSION: In the majority of cases, subjects had an undiagnosed primary cancer at the time the metastatic jaw disease presented. The most common site of origin of the primary cancer was the breast, when primary sites were considered independent of sex. CLINICAL IMPLICATIONS: Patients with metastatic disease in the jaws may have innocuous dental symptoms, such as pulpal or periodontal pain; therefore, clinicians will play a significant role in diagnosing the life-threatening disease. PMID- 17138712 TI - Diagnostic tools for early caries detection. AB - BACKGROUND: Management of dental caries demands early detection of carious lesions. This article provides an overview of the state-of-the-art methodologies for the detection and assessment of early carious lesions. TYPES OF STUDIES REVIEWED: The authors reviewed MEDLINE for available literature on new caries detection methodology and tools, using terms such as "early caries detection," "fluorescence" and "transillumination." Their review was not a systematic review of the literature. They included in their review in vitro, in situ, in vivo and clinical studies, as well as position papers, editorials and consensus conferences statements published in English. RESULTS: Each early caries detection tool has advantages and disadvantages; some perform better on certain surfaces than others. Therefore, their performance threshold and the operator's influence on performance must be considered. Not all methods accurately detect early lesions, and false positives and false negatives may occur. Detecting early lesions in combination with assessing activity status is essential for establishing the prognosis and threshold required for preventive intervention. Clinically useful tools to help make such decisions are under development. CLINICAL IMPLICATIONS: Early caries detection methods should be an adjunct to clinical decision making, supporting preventive treatment planning in conjunction with caries risk assessment but not justifying premature restorative intervention. PMID- 17138713 TI - The significance of needle bevel orientation in achieving a successful inferior alveolar nerve block. AB - BACKGROUND: The authors conducted a prospective, randomized, single-blinded, crossover study comparing the degree of pulpal anesthesia achieved with the use of a conventional inferior alveolar nerve (IAN) block administered with the needle bevel oriented away from the mandibular ramus or toward the mandibular ramus. METHODS: Fifty-one blinded subjects randomly received an IAN block injection administered with a 27-gauge needle; the needle bevel was oriented away from the mandibular ramus or oriented toward the mandibular ramus at appointments spaced at least one week apart, in a crossover design. The authors used a pulp tester to test molars, premolars and central and lateral incisors for anesthesia in four-minute cycles for 60 minutes. They considered anesthesia to be successful when two consecutive 80 readings (the maximum output on the pulp tester) were obtained within 15 minutes, and the 80 reading was sustained continuously for 60 minutes. RESULTS: When the needle bevel was oriented away from the mandibular ramus, successful pulpal anesthesia from the central incisor to the second molar was achieved in 24 to 90 percent of patients. When the needle bevel was oriented toward the mandibular ramus, successful pulpal anesthesia was achieved in 14 to 92 percent of patients. The results showed no significant difference between the two needle bevel orientations. CONCLUSION: The authors concluded that using a 27 gauge needle with the bevel oriented away from the mandibular ramus was similar to using the same needle with the bevel oriented toward the mandibular ramus to administer successful IAN blocks in adults. CLINICAL IMPLICATIONS: For IAN blocks administered with a 27-gauge needle, positioning the needle bevel away or toward the mandibular ramus does not affect anesthetic success. PMID- 17138714 TI - Alcohol screening in dental patients: the prevalence of hazardous drinking and patients' attitudes about screening and advice. AB - BACKGROUND: Because heavy drinking is a risk factor for oral cancer, dentists should screen patients for alcohol use. The authors investigated heavy drinking in dental patients and patients' attitudes about alcohol screening. METHODS: A convenience sample of 408 patients attending an emergency walk-in dental clinic served as subjects. Patients completed the Alcohol Use Disorders Identification Test-C (AUDIT-C), a three-item alcohol screening test, and an opinion survey regarding attitudes about the acceptability of alcohol screening and counseling by dentists. RESULTS: One in four patients had positive screening results for heavy alcohol use. The majority of subjects (> 75 percent) were in support of dentists' inquiries and advice about alcohol use. Age, sex and drinking status were not predictive of patients' opinions about alcohol screening. CONCLUSIONS: One hundred three of the dental patients exhibited evidence of hazardous alcohol consumption, a risk factor for oropharyngeal cancer. The majority of patients reported that they would readily accept alcohol screening and alcohol counseling by dentists. CLINICAL IMPLICATIONS: Because studies have shown that some dentists hesitate to screen for alcohol use because of a belief that screening is unacceptable to patients, these results may encourage a change in practice. PMID- 17138715 TI - The radiographic outcomes of direct pulp-capping procedures performed by dental students: a retrospective study. AB - BACKGROUND: The decision between pulp capping and root canal therapy after pulp exposure is a clinical issue. The aim of the authors' study was to evaluate the outcome of direct pulp-capping procedures performed by dental students. METHODS: The authors followed the treatment outcomes of 193 patients with 204 pulp exposures with direct pulp capping. They determined the outcome of pulp capping radiographically using periapical radiographs taken at least three years after pulp exposure. The outcome was considered as successful if the tooth was present and not associated with periapical radiolucency or root canal treatment; otherwise, the outcome was considered as being a failure. RESULTS: Overall, the success rate of pulp capping was 59.3 percent. The success was associated more with mechanical exposure than with carious exposure (92.2 versus 33.3 percent) (P < .001), more with permanent restoration than with temporary restoration (80.8 versus 47.3 percent) (P < .001) and more with class I occlusal restoration (83.8 percent) than with proximal multiple surface restorations (Class II, 56.1 percent; Class III, 58.8 percent; mesial-occlusal-distal, 28.6 percent) (P = .009). Patients' age, sex, and tooth location and position had no significant effect on the outcome (P > .05). CONCLUSION: The success rate of direct pulp capping was 92.2 percent with mechanical exposure and 33.3 percent with carious exposure. CLINICAL IMPLICATIONS: Direct pulp capping is recommended after mechanical exposure with immediate placement of permanent restoration, while root canal therapy would be the choice of treatment if the exposure was due to caries. PMID- 17138716 TI - Considerations in asset allocation: investing wisely. AB - BACKGROUND: In this article, the author reviews the steps an investor should take to make sound asset allocation decisions and offers guidelines on assembling a portfolio that helps meet the investor's needs. CONCLUSIONS: The most important decision in structuring a broadly diversified investment portfolio is the allocation among and within the three major asset classes: stocks, bonds and short-term reserves. In selecting the appropriate mix, the investor should weigh four interrelated factors-investment objective, time horizon, risk tolerance and personal financial situation. PRACTICE IMPLICATIONS: A well-diversified investment portfolio and a disciplined savings program are important factors in reaching financial goals. PMID- 17138717 TI - Using mouthguards to reduce the incidence and severity of sports-related oral injuries. AB - BACKGROUND: This report reviews the available literature on the types and properties of athletic mouthguards, current fabrication methods and the role of mouth protectors in reducing the incidence and severity of sports-related oral injuries. OVERVIEW: For more than 50 years, the American Dental Association has promoted the protective value of wearing properly fitted mouthguards while participating in athletic or recreational activities that carry a risk of dental injury. Safety is essential to maintaining oral health, and a properly fitted mouthguard can minimize the risks of sustaining oral injuries during participation in sports. CONCLUSIONS: The dental literature supports the use and protective value of mouthguards in reducing sports-related injuries to the teeth and soft tissues. Dentists are encouraged to educate patients regarding the risks of oral injury in sports, fabricate properly fitted mouthguards, and provide appropriate guidance on mouthguard types and their protective properties, costs and benefits. Further studies addressing the effectiveness of currently available mouthguard types and population-based interventions for reducing oral injuries are recommended. CLINICAL IMPLICATIONS: Participants in sporting and recreational activities are often susceptible to oral injury. To reduce the incidence and severity of sports-related oral trauma, the use of a properly fitted mouthguard is recommended in any athletic or recreational activity that carries a risk of injury. PMID- 17138718 TI - Are veneers conservative treatment? PMID- 17138719 TI - Improving case acceptance: a new paradigm for treatment plan presentations. PMID- 17138721 TI - For the dental patient ... Protecting teeth with mouthguards. PMID- 17138722 TI - Effect of CCK-1 receptor blockade on ghrelin and PYY secretion in men. AB - Cholecystokinin (CCK), peptide YY (PYY), and ghrelin have been proposed to act as satiety hormones. CCK and PYY are stimulated during meal intake by the presence of nutrients in the small intestine, especially fat, whereas ghrelin is inhibited by eating. The sequence of events (fat intake followed by fat hydrolysis and CCK release) suggests that this process is crucial for triggering the effects. The aim of this study was therefore to investigate whether CCK mediated the effect of intraduodenal (ID) fat on ghrelin secretion and PYY release via CCK-1 receptors. Thirty-six male volunteers were studied in three consecutive, randomized, double blind, cross-over studies: 1) 12 subjects received an ID fat infusion with or without 120 mg orlistat, an irreversible inhibitor of gastrointestinal lipases, compared with vehicle; 2) 12 subjects received ID long-chain fatty acids (LCF), ID medium-chain fatty acids (MCF), or ID vehicle; and 3) 12 subjects received ID LCF with and without the CCK-1 receptor antagonist dexloxiglumide (Dexlox) or ID vehicle plus intravenous saline (placebo). ID infusions were given for 180 min. The effects of these treatments on ghrelin concentrations and PYY release were quantified. Plasma hormone concentrations were measured in regular intervals by specific RIA systems. We found the following results. 1) ID fat induced a significant inhibition in ghrelin levels (P < 0.01) and a significant increase in PYY concentrations (P < 0.004). Inhibition of fat hydrolysis by orlistat abolished both effects. 2) LCF significantly inhibited ghrelin levels (P < 0.02) and stimulated PYY release (P < 0.008), whereas MCF were ineffective compared with controls. 3) Dexlox administration abolished the effect of LCF on ghrelin and on PYY. ID fat or LCF significantly stimulated plasma CCK (P < 0.006 and P < 0.004) compared with saline. MCF did not stimulate plasma CCK release. In summary, fat hydrolysis is essential to induce effects on ghrelin and PYY through the generation of LCF, whereas MCF are ineffective. Furthermore, LCF stimulated plasma CCK release, suggesting that peripheral CCK is the mediator of these actions. The CCK-1 receptor antagonist Dexlox abolished the effect of ID LCF, on both ghrelin and PYY. Generation of LCF through hydrolysis of fat is a critical step for fat-induced inhibition of ghrelin and stimulation of PYY in humans; the signal is mediated via CCK release and CCK-1 receptors. PMID- 17138723 TI - High salt intake reduces endothelium-dependent dilation of mouse arterioles via superoxide anion generated from nitric oxide synthase. AB - In skeletal muscle arterioles of normotensive rats fed a high salt diet, the bioavailability of endothelium-derived nitric oxide (NO) is reduced by superoxide anion. Because the impact of dietary salt on resistance vessels in other species is largely unknown, we investigated endothelium-dependent dilation and oxidant activity in spinotrapezius muscle arterioles of C57BL/6J mice fed normal (0.45%, NS) or high salt (7%, HS) diets for 4 wk. Mean arterial pressure in HS mice was not different from that in NS mice, but the magnitude of arteriolar dilation in response to different levels of ACh was 42-57% smaller in HS mice than in NS mice. Inhibition of nitric oxide synthase (NOS) with N(G) monomethyl L-arginine (L-NMMA) significantly reduced resting diameters and reduced responses to ACh (by 45-63%) in NS mice but not in HS mice. Arteriolar wall oxidant activity, as assessed by tetranitroblue tetrazolium reduction or hydroethidine oxidation, was greater in HS mice than in NS mice. Exposure to the superoxide scavenger 2,2,6,6 tetramethylpiperidine-N-oxyl (TEMPO) + catalase reduced this oxidant activity to normal and restored normal arteriolar responsiveness to ACh in HS mice but had no effect in NS mice. L-NMMA also restored arteriolar oxidant activity to normal in HS mice. ACh further increased arteriolar oxidant activity in HS mice but not in NS mice, and this effect was prevented with L-NMMA. These data suggest that a high salt diet promotes increased generation of superoxide anion from NOS in the murine skeletal muscle microcirculation, thus impairing endothelium-dependent dilation through reduced NO bioavailability. PMID- 17138724 TI - Thermogenic side effects to migratory predisposition in shorebirds. AB - In the calidrine sandpiper red knot (Calidris canutus), the weeks preceding takeoff for long-distance migration are characterized by a rapid increase in body mass, largely made up of fat but also including a significant proportion of lean tissue. Before takeoff, the pectoral muscles are known to hypertrophy in preparation for endurance flight without any specific training. Because birds facing cold environments counterbalance heat loss through shivering thermogenesis, and since pectoral muscles represent a large proportion of avian body mass, we asked the question whether muscle hypertrophy in preparation for long-distance endurance flight would induce improvements in thermogenic capacity. We acclimated red knots to different controlled thermal environments: 26 degrees C, 5 degrees C, and variable conditions tracking outdoor temperatures. We then studied within-individual variations in body mass, pectoral muscle size (measured by ultrasound), and metabolic parameters [basal metabolic rate (BMR) and summit metabolic rate (M(sum))] throughout a 3-mo period enclosing the migratory gain and loss of mass. The gain in body mass during the fattening period was associated with increases in pectoral muscle thickness and thermogenic capacity independent of thermal acclimation. Regardless of their thermal treatment, birds showing the largest increases in body mass also exhibited the largest increases in M(sum). We conclude that migratory fattening is accompanied by thermoregulatory side effects. The gain of body mass and muscle hypertrophy improve thermogenic capacity independent of thermal acclimation in this species. Whether this represents an ecological advantage depends on the ambient temperature at the time of fattening. PMID- 17138725 TI - Sex differences in postexercise esophageal and muscle tissue temperature response. AB - Factors associated with blood pressure regulation during recovery from exercise dramatically influence core temperature regulation. However, it is unknown whether sex-related differences in postexercise hemodynamics affect core and muscle temperature response. Sixteen participants (8 males, 8 females) completed an incremental isotonic test on a Kin-Com isokinetic apparatus to determine their activity-specific peak oxygen consumption during bilateral knee extensions (Vo(2)(sp)). On a separate day, participants performed 15 min of isolated bilateral knee extensions at a moderate (60% Vo(2)(sp)) exercise intensity followed by a 90-min recovery. Esophageal temperature (T(es)), mean arterial pressure (MAP), muscle temperature at four depths in the active vastus medialis (T(VM)) and three depths in the inactive triceps brachii (T(TB)) were measured concurrently with sweat rate and cutaneous vascular conductance (CVC). Relative to the preexercise resting T(es) of 36.7 degrees C (SD 0.1), between 10 and 50 min of recovery T(es) was 0.19 degrees C (SD 0.02) higher for females than males (P = 0.037). All measurements of T(VM) (0.036 > P > 0.014) and T(TB) (0.048 > P > 0.008) were higher for females during the initial 30 min of recovery by between 0.46 degrees C and 0.64 degrees C for T(VM) and by between 0.53 degrees C and 0.70 degrees C for T(TB). In parallel, females showed a 5 to 7 mmHg greater reduction in MAP during recovery relative to males (P = 0.002) and a significantly lower CVC (P = 0.020) and sweat rate (P = 0.034). Therefore, it is concluded that females demonstrate a greater and more prolonged elevation in postexercise esophageal temperature and active and inactive muscle temperatures, which is paralleled by a greater postexercise hypotensive response. PMID- 17138726 TI - Disruption of adenosinergic modulation of ventilation at rest and during hypercapnia by neonatal caffeine in young rats: role of adenosine A(1) and A(2A) receptors. AB - Caffeine is commonly used to treat respiratory instabilities related to prematurity. However, the role of adenosinergic modulation and the potential long term effects of neonatal caffeine treatment (NCT) on respiratory control are poorly understood. To address these shortcomings, we tested the following hypotheses: 1) adenosine A(1)- and A(2A)-receptor antagonists modulate respiratory activity at rest and during hypercapnia; 2) NCT has long-term consequences on adenosinergic modulation of respiratory control. Rat pups received by gavage either caffeine (15 mg/kg) or water (control) once a day from postnatal days 3 to 12. At day 20, rats received intraperitoneal injection with vehicle, DPCPX (A(1) antagonist, 4 mg/kg), or ZM-241385 (A(2A) antagonist, 1 mg/kg) before plethysmographic measurements of resting ventilation, hypercapnic ventilatory response (5% CO(2)), and occurrence of apneas in freely behaving rats. In controls, data show that A(2A), but not A(1), antagonist decreased resting ventilation by 31% (P = 0.003). A(1) antagonist increased the hypercapnic response by 60% (P < 0.001), whereas A(2A) antagonist increased the hypercapnic response by 42% (P = 0.033). In NCT rats, A(1) antagonist increased resting ventilation by 27% (P = 0.02), but the increase of the hypercapnic response was blunted compared with controls. A(1) antagonist enhanced the occurrence of spontaneous apneas in NCT rats only (P = 0.005). Finally, A(2A) antagonist injected in NCT rats had no effect on ventilation. These data show that hypercapnia activates adenosinergic pathways, which attenuate responsiveness (and/or sensitivity) to CO(2) via A(1) receptors. NCT elicits developmental plasticity of adenosinergic modulation, since neonatal caffeine persistently decreases ventilatory sensitivity to adenosine blockers. PMID- 17138727 TI - Serotonergic mechanisms of the lateral parabrachial nucleus in renal and hormonal responses to isotonic blood volume expansion. AB - This study investigated the involvement of serotonergic mechanisms of the lateral parabrachial nucleus (LPBN) in the control of sodium (Na+) excretion, potassium (K+) excretion, and urinary volume in unanesthetized rats subjected to acute isotonic blood volume expansion (0.15 M NaCl, 2 ml/100 g of body wt over 1 min) or control rats. Plasma oxytocin (OT), vasopressin (VP), and atrial natriuretic peptide (ANP) levels were also determined in the same protocol. Male Wistar rats with stainless steel cannulas implanted bilaterally into the LPBN were used. In rats treated with vehicle in the LPBN, blood volume expansion increased urinary volume, Na+ and K+ excretion, and also plasma ANP and OT. Bilateral injections of serotonergic receptor antagonist methysergide (1 or 4 microg/200 etal) into the LPBN reduced the effects of blood volume expansion on increased Na+ and K+ excretion and urinary volume, while LPBN injections of serotonergic 5 HT(2a)/HT(2c) receptor agonist, 2.5-dimetoxi-4-iodoamphetamine hydrobromide (DOI; 1 or 5 microg/200 etal) enhanced the effects of blood volume expansion on Na+ and K+ excretion and urinary volume. Methysergide (4 microg) into the LPBN decreased the effects of blood volume expansion on plasma ANP and OT, while DOI (5 microg) increased them. The present results suggest the involvement of LPBN serotonergic mechanisms in the regulation of urinary sodium, potassium and water excretion, and hormonal responses to acute isotonic blood volume expansion. PMID- 17138728 TI - Interleukin-1 receptor antagonist as a modulator of gender differences in the febrile response to lipopolysaccharide in rats. AB - Febrile responses to bacterial pathogens are attenuated near term of pregnancy in several mammalian species. It is unknown, however, whether this reflects a fundamental physiological adaptation of female rats or whether it is specific to pregnancy. The aims of this study therefore were 1) to determine whether febrile responses to the bacterial endotoxin lipopolysaccharide (LPS) are attenuated in female vs. male rats and, if so, to identify possible mechanisms involved in modulating this and 2) to assess whether plasma concentrations of the anti inflammatory cytokine, interleukin-1 receptor antagonist (IL-1ra), an important regulator of fever, are dependent on the physiological state of the female and could therefore be involved in modulating febrile responses. We found febrile responses were attenuated in cycling female vs. male rats and also in near-term pregnant dams vs. cycling females after intraperitoneal injection of LPS (0.05 mg/kg). Plasma levels of IL-1ra were significantly greater in female rats after injection of LPS, particularly during pregnancy, than in males. This was accompanied by attenuated levels of hypothalamic IL-1beta and cyclooxygenase-2 mRNA, two key mediators of the febrile response, in female rats. Furthermore, increasing plasma levels of IL-1ra in male rats by intraperitoneal administration of the recombinant antagonist attenuated hypothalamic mRNA levels of these mediators after LPS. These data suggest that there is a fundamental difference in febrile response to LPS between the genders that is likely regulated by IL-1ra. This may be an important mechanism that protects the developing fetus from potentially deleterious consequences of maternal fever during pregnancy. PMID- 17138729 TI - Antenatal antioxidant prevents adult hypertension, vascular dysfunction, and microvascular rarefaction associated with in utero exposure to a low-protein diet. AB - Developmental programming of hypertension is associated with vascular dysfunction characterized by impaired vasodilatation to nitric oxide, exaggerated vasoconstriction to ANG II, and microvascular rarefaction appearing in the neonatal period. Hypertensive adults have indices of increased oxidative stress, and newborns that were nutrient depleted during fetal life have decreased antioxidant defenses and increased susceptibility to oxidant injury. To test the hypothesis that oxidative stress participates in early life programming of hypertension, vascular dysfunction, and microvascular rarefaction associated with maternal protein deprivation, pregnant rats were fed a normal, low protein (LP), or LP plus lazaroid (lipid peroxidation inhibitor) isocaloric diet from the day of conception until delivery. Lazaroid administered along with the LP diet prevented blood pressure elevation, enhanced vasomotor response to ANG II, impaired vasodilatation to sodium nitroprusside, and microvascular rarefaction in adult offspring. Liver total glutathione was significantly decreased in LP fetuses, and kidney eight-isoprostaglandin F2alpha (8-isoPGF(2alpha)) levels were significantly increased in adult LP offspring; these modifications were prevented by lazaroid. Renal nitrotyrosine abundance and blood levels of 1,4 dihydroxynonene and 4-hydroxynonenal-protein adducts were not modified by antenatal diet exposure. This study shows in adult offspring of LP-fed dams prevention of hypertension, vascular dysfunction, microvascular rarefaction, and of an increase in indices of oxidative stress by the administration of lazaroid during gestation. Lazaroid also prevented the decrease in antioxidant glutathione levels in fetuses, suggesting an antenatal mild oxidative stress in offspring of LP-fed dams. These studies support the concept that perinatal oxidative insult can lead to permanent alterations in the cardiovascular system development. PMID- 17138731 TI - Receptor-mediated tumor targeting with radiolabeled peptides: there is more to it than somatostatin analogs. PMID- 17138732 TI - Usefulness of hybrid SPECT/CT in 99mTc-HMPAO-labeled leukocyte scintigraphy for bone and joint infections. AB - White blood cell scanning with (99m)Tc-hexamethylpropylene amine oxime (HMPAO) has proven highly sensitive and specific in the diagnosis and follow-up of patients with suspected osteomyelitis. The aim of this prospective study was to evaluate the usefulness of SPECT and transmission CT performed simultaneously using a hybrid imaging device for the functional anatomic mapping of bone and joint infections. METHODS: (99m)Tc-HMPAO scintigraphy was performed on 28 consecutive patients: 15 with suspected bone infection (group 1) and 13 with suspected orthopedic implant infection (group 2). Planar scans were acquired 30 min, 4 h, and 24 h after injection. SPECT/CT was obtained 6 h after tracer injection, using a dual-head gamma-camera coupled with a low-power x-ray tube. In all patients, scintigraphic results were matched with the results of surgery or cultures and of clinical follow-up. RESULTS: (99m)Tc-HMPAO scintigraphy was true positive for infection in 18 of 28 patients (for a total of 21 sites of uptake) and true-negative in 10 of 28 subjects. SPECT/CT provided an accurate anatomic localization of all positive foci. With regard to the final diagnosis, SPECT/CT added a significant clinical contribution in 10 of 28 patients (35.7%). In fact, SPECT/CT differentiated soft-tissue from bone involvement both in patients with osteomyelitis and in patients with orthopedic implants, allowed correct diagnosis of osteomyelitis in patients with structural alterations after trauma, and identified synovial infection without prosthesis involvement in patients with a knee implant. CONCLUSION: Our results indicate that SPECT/CT performed using a hybrid device can improve imaging with (99m)Tc-HMPAO-labeled leukocytes in patients with suspected osteomyelitis by providing accurate anatomic localization and precise definition of the extent of infection. PMID- 17138733 TI - Smoking cessation normalizes coronary endothelial vasomotor response assessed with 15O-water and PET in healthy young smokers. AB - Cigarette smoking is one of the risk factors of cardiovascular diseases and is related to abnormal peripheral and coronary vascular vasomotion. Coronary vascular endothelial dysfunction is caused by chronic smoking in smokers without epicardial coronary artery stenosis. The coronary endothelial vasomotion abnormality is restored by interventions such as l-arginine or vitamin C infusion. However, to our knowledge, the effect of smoking cessation on coronary vasomotor response has not been elucidated. Therefore, the aim of this study was to assess the effect of smoking cessation on coronary vasomotor response by quantitative myocardial blood flow (MBF) measurement using (15)O-water and PET. METHODS: Fifteen young smokers (Brinkman index > 100; mean age +/- SD, 26 +/- 4 y) with no evidence of heart disease or cardiovascular risk factors, except for smoking, and age-matched nonsmokers (n = 12) were enrolled in this study. MBF was measured at rest, during the cold pressor test (CPT), before and at 1 and 6 mo after smoking cessation. In addition, MBF measurement during adenosine triphosphate (ATP) infusion was performed before and at 6 mo after smoking cessation. In nonsmokers, MBF was measured at rest, during ATP infusion, and during the CPT. RESULTS: MBF at rest and during ATP infusion did not differ between smokers and nonsmokers (0.73 +/- 0.12 vs. 0.80 +/- 0.15 mL/g/min and 3.15 +/- 1.43 vs. 3.69 +/- 0.76 mL/g/min, respectively; P = not significant). In contrast, MBF during the CPT in smokers was lower than that in nonsmokers (0.90 +/- 0.19 vs. 1.12 +/- 0.28 mL/g/min; P < 0.05). There was no significant difference in MBF either at rest or during ATP infusion between before and after smoking cessation, but MBF during the CPT increased at 1 mo in comparison with before cessation of smoking (0.90 +/- 0.19 vs. 1.02 +/- 0.22 mL/g/min; P < 0.01). An improvement of MBF response to the CPT was preserved at 6 mo after smoking cessation. CONCLUSION: Coronary vasomotor abnormality assessed by MBF response to the CPT was improved at 1 mo after smoking cessation. These findings indicate that coronary endothelial dysfunction may be reversible within 1 mo after smoking cessation in healthy young smokers. PMID- 17138734 TI - Lack of correlation of hypoxic cell fraction and angiogenesis with glucose metabolic rate in non-small cell lung cancer assessed by 18F-Fluoromisonidazole and 18F-FDG PET. AB - PET offers a noninvasive means to assess neoplasms, in view of its sensitivity and accuracy in staging tumors and potentially in monitoring treatment response. The aim of this study was to evaluate newly diagnosed non-small cell lung cancer (NSCLC) for the presence of hypoxia, as indicated by the uptake of (18)F Fluoromisonidazole ((18)F-FMISO), and to examine the relationship of hypoxia to the uptake of (18)F-FDG, microvessel density, and other molecular markers of hypoxia. METHODS: Twenty-one patients with suspected or biopsy-proven NSCLC were enrolled prospectively in this study. All patients had PET studies with (18)F FMISO and (18)F-FDG. Seventeen patients subsequently underwent surgery, with analysis performed for tumor markers of angiogenesis and hypoxia. RESULTS: In the 17 patients with resectable NSCLC (13 men, 4 women; age range, 51-77 y), the mean (18)F-FMISO uptake in tumor was significantly lower than that of (18)F-FDG uptake (P < 0.0001) and showed no correlation with (18)F-FDG uptake (r = 0.26). The mean (95% confidence interval [CI]) (18)F-FMISO SUV(max) (maximum standardized uptake value) was 1.20 [0.95-1.45] compared with the mean [95% CI] (18)F-FDG SUV(max) of 5.99 [4.62-7.35]. The correlation between (18)F-FMISO uptake, (18)F-FDG uptake, and tumor markers of hypoxia and angiogenesis was poor. A weakly positive correlation between (18)F-FMISO and (18)F-FDG uptake and Ki67 was found. CONCLUSION: The hypoxic cell fraction of primary NSCLC is consistently low, and there is no significant correlation in NSCLC between hypoxia and glucose metabolism in NSCLC assessed by (18)F-FDG. These findings have direct implications in understanding the role of angiogenesis and hypoxia in NSCLC biology. PMID- 17138735 TI - 186Re-HEDP in the treatment of patients with inoperable osteosarcoma. AB - The aim of this study was to examine the safety and efficacy of (186)Re hydroxyethylidene diphosphonate (HEDP) as an adjuvant to external-beam radiotherapy (EBRT) in the treatment of patients with osteosarcoma. METHODS: Thirteen patients (9 male, 4 female; age range, 12-42 y) were treated with combination chemotherapy (standard U.K. protocol) and (186)Re-HEDP therapy (18.5 MBq/kg, intravenously), followed by EBRT. A full blood count; liver function test; and measurements of urea and electrolytes, glomerular filtration rate, and left ventricular function were performed on all patients before and after therapy. Tumor volume and composition were obtained from CT or MRI data. Dosimetric calculations were performed using the MIRD formalism. RESULTS: Of the 13 patients, 1 is still under follow-up. The median survival time was 36 mo (range, 12-216 mo) from diagnosis and 5 mo (range, 1-60 mo) from the last (186)Re HEDP treatment. The mean tumor dose delivered with (186)Re-HEDP was calculated to be 5.8 Gy (range, 0.5-16 Gy). CT and MRI revealed the tumors to have a complex structure, comprising "ossified," "partially calcified," and "soft-tissue" components. Posttherapy scans showed a heterogeneous distribution of (186)Re-HEDP in the tumor mass: Although the "soft-tissue" component showed minimal uptake of the therapeutic dose, the "ossified component" showed intense uptake. The 3 long term survivors in whom tumor sterilization was achieved received calculated mean tumor doses in the range of 2.0-3.1 Gy, which was believed to be an underestimate of the actual tumor doses delivered. CONCLUSION: This study indicates that a simple approach to tumor dosimetry based on mean tumor dose is inappropriate because it may underestimate the dose delivered to these heterogeneous tumors. The data also indicate that EBRT combined with a standard dose of 18.5 MBq/kg of (186)Re-HEDP does not provide a sufficient dose to achieve tumor sterilization. A dose estimation technique is required that is based on the determination of tumor dose at the individual voxel level and that is able to represent the heterogeneous uptake observed in these complex tumor structures with highly nonuniform composition. This, coupled with individualized dose escalation, may then achieve the goal of tumor sterilization. PMID- 17138736 TI - Correction of head movement on PET studies: comparison of methods. AB - Head movement presents a continuing problem in PET studies. Head restraint minimizes movement but is unreliable, resulting in the need to develop alternative strategies. These include frame-by-frame (FBF) realignment or use of motion tracking (MT) during the scan to realign PET acquisition data. Here we present a comparative analysis of these 2 methods of motion correction. METHODS: Eight volunteers were examined at rest using (11)C-raclopride PET with the radioligand administered as a bolus followed by constant infusion to achieve steady state. Binding potential (BP) was estimated using the ratio method during 2 periods of the scan at steady state. Head movement was compensated by using coregistration between frames (FBF) and 3 methods using MT measurements of head position acquired with a commercially available optical tracking system. RESULTS: All methods of realignment improved test-retest reliability and noise characteristics of the raw data, with important consequences for the power to detect small changes in radiotracer binding, and the potential to reduce false positive and false-negative results. MT methods were superior to FBF realignment using coregistration on some indices. CONCLUSION: Such methods have considerable potential to improve the reliability of PET data with important implications for the numbers of volunteers required to test hypotheses. PMID- 17138737 TI - Osteoporosis redux. PMID- 17138738 TI - NEMA NU 2-2001 performance measurements of an LYSO-based PET/CT system in 2D and 3D acquisition modes. AB - The National Electrical Manufacturers Association (NEMA) NU 2-2001 performance measurements were conducted on the Discovery RX, a whole-body PET/CT system under development by GE Healthcare. The PET scanner uses 4.2 x 6.3 x 30 mm lutetium yttrium orthosilicate (LYSO) crystals grouped in 9 x 6 blocks. There are 24 rings with 630 crystals per ring and the ring diameter is 88.6 cm. The transaxial and axial fields of view are 70.0 and 15.7 cm, respectively. The scanner has retractable septa and can operate in both 2-dimensional (2D) and 3-dimensional (3D) modes. 2D acquisitions use ring differences of +/-4 for direct and +/-5 for cross slices; 3D acquisitions use a ring difference of 23. The coincident window width is 6.5 ns and the energy window is 425-650 keV. Other than the detectors, the system uses the same hardware and software as a Discovery ST. The CT scanner is a 16-slice LightSpeed; the performance characteristics of the CT component are not included herein. METHODS: Performance measurements of sensitivity, spatial resolution, image quality, scatter fraction and counting rate performance, and image quality were obtained using NEMA methodology. RESULTS: The system sensitivity in 2D and 3D was measured as 1.7 cps/kBq and 7.3 cps/kBq, respectively. The transaxial resolution for 2D (3D) was 5.1 mm full width at half maximum (FWHM) (5.0 mm) at 1 cm from gantry center and the radial and tangential resolutions were 5.9 mm (5.9 mm) and 5.1 mm (5.2 mm) at 10 cm, respectively. The axial resolution for 2D (3D) was 4.8 mm FWHM (5.8 mm) and 6.3 mm (6.5 mm) at 1 cm and 10 cm from gantry center, respectively. The scatter fraction was 13.1% and 31.8% in 2D and 3D. The peak noise equivalent count rate (NECR) was 155 kcps at 92.1 kBq/mL in 2D and 117.7 kcps at 21.7 kBq/mL in 3D for a noise-free estimation of randoms. The contrast of the 22, 17, 13, and 10 mm hot spheres in the image quality phantom in 2D (3D) were 74.6% (72.4%), 56.7% (59.5%), 46.2% (44.6%), and 17.9% (18.0%), respectively. CONCLUSION: The Discovery RX is a scanner that possesses high NECR, low scatter fraction, and good spatial resolution characteristics. PMID- 17138739 TI - Simultaneous acquisition of multislice PET and MR images: initial results with a MR-compatible PET scanner. AB - PET and MRI are powerful imaging techniques that are largely complementary in the information they provide. We have designed and built a MR-compatible PET scanner based on avalanche photodiode technology that allows simultaneous acquisition of PET and MR images in small animals. METHODS: The PET scanner insert uses magnetic field-insensitive, position-sensitive avalanche photodiode (PSAPD) detectors coupled, via short lengths of optical fibers, to arrays of lutetium oxyorthosilicate (LSO) scintillator crystals. The optical fibers are used to minimize electromagnetic interference between the radiofrequency and gradient coils and the PET detector system. The PET detector module components and the complete PET insert assembly are described. PET data were acquired with and without MR sequences running, and detector flood histograms were compared with the ones generated from the data acquired outside the magnet. A uniform MR phantom was also imaged to assess the effect of the PET detector on the MR data acquisition. Simultaneous PET and MRI studies of a mouse were performed ex vivo. RESULTS: PSAPDs can be successfully used to read out large numbers of scintillator crystals coupled through optical fibers with acceptable performance in terms of energy and timing resolution and crystal identification. The PSAPD LSO detector performs well in the 7-T magnet, and no visible artifacts are detected in the MR images using standard pulse sequences. CONCLUSION: The first images from the complete system have been successfully acquired and reconstructed, demonstrating that simultaneous PET and MRI studies are feasible and opening up interesting possibilities for dual-modality molecular imaging studies. PMID- 17138740 TI - Lung toxicity in radioiodine therapy of thyroid carcinoma: development of a dose rate method and dosimetric implications of the 80-mCi rule. AB - Based on an extensive dataset analyzed by Benua et al., a whole-body retention threshold of 2.96 GBq (80 mCi) at 48 h has been used to limit the radioactivity of (131)I administered to thyroid cancer patients with diffuse pulmonary metastases. In this work, the 80-mCi activity retention limit is used to derive lung-absorbed doses and dose rates. The resulting dose-rate-based limits make it possible to account for patient-specific differences in lung geometry. This is particularly important, for example, in pediatric patients exhibiting diffuse lung metastases. The approach also highlights the impact of altered radioiodine kinetics as seen with recombinant human thyroid-stimulating hormone. METHODS: The dose-rate constraint (DRC) was defined as the absorbed dose rate to the lungs of the adult female reference phantom when 80 mCi of (131)I are in the body and 90% of this is uniformly distributed in the lungs. With this definition, the 80-mCi rule was generalized by calculating the activity required to yield a dose rate equal to DRC using lung-to-lung S factor values corresponding to different reference phantoms. RESULTS: A DRC value of 43.6 cGy/h was obtained. Applying this DRC to the adult male phantom and to the phantom of a 15-y-old yields equivalent 48-h activity limits of 3.72 GBq (101 mCi) and 2.45 GBq (66.2 mCi), respectively. Depending on model parameters, the absorbed doses to lungs ranged from 57 to 112 Gy; the photon-only portion, which better reflects the dose to normal lung parenchyma, ranged from 4.9 to 55 Gy. CONCLUSION: A dose-rate-based version of the 80-mCi rule is derived and used to demonstrate application of this rule to pediatric patients and to adult male patients. The implications of the 80 mCi rule are also examined. The assumption of uniform energy deposition in the lungs leads to substantially overestimated absorbed doses. Severe radiation induced lung toxicity, expected at normal lung absorbed doses of 25-27 Gy, is avoided, probably because most of the local electron dose is delivered to tumor tissue instead of to normal lung parenchyma. The possibility of using a DRC to adjust treatment for different clinical situations is illustrated. The analysis suggests that a dosimetry-based approach will be particularly important in the treatment of patients with lung metastases when a recombinant human thyroid stimulating hormone protocol is used. PMID- 17138742 TI - Predicting chemotherapy response to paclitaxel with 18F-Fluoropaclitaxel and PET. AB - Paclitaxel is used as a chemotherapy drug for the treatment of various malignancies, including breast, ovarian, and lung cancers. To evaluate the potential of a noninvasive prognostic tool for specifically predicting the resistance of tumors to paclitaxel therapy, we examined the tumoral uptake of (18)F-fluoropaclitaxel ((18)F-FPAC) in mice bearing human breast cancer xenografts by using small-animal-dedicated PET and compared (18)F-FPAC uptake with the tumor response to paclitaxel treatment. METHODS: PET data were acquired after tail vein injection of approximately 9 MBq of (18)F-FPAC in anesthetized nude mice bearing breast cancer xenografts. Tracer uptake in reconstructed images was quantified by region-of-interest analyses and compared with the tumor response, as measured by changes in tumor volume, after treatment with paclitaxel. RESULTS: Mice with tumors that progressed demonstrated lower tumoral uptake of (18)F-FPAC than mice with tumors that did not progress or that regressed (r = 0.55, P < 0.02; n = 19), indicating that low (18)F-FPAC uptake was a significant predictor of chemoresistance. Conversely, high (18)F-FPAC uptake predicted tumor regression. This relationship was found for mice bearing xenografts from cell lines selected to be either sensitive or intrinsically resistant to paclitaxel in vitro. CONCLUSION: PET data acquired with (18)F-FPAC suggest that this tracer holds promise for the noninvasive quantification of its distribution in vivo in a straightforward manner. In combination with approaches for examining other aspects of resistance, such quantification could prove useful in helping to predict subsequent resistance to paclitaxel chemotherapy of breast cancer. PMID- 17138741 TI - Lung dosimetry for radioiodine treatment planning in the case of diffuse lung metastases. AB - The lungs are the most frequent sites of distant metastasis in differentiated thyroid carcinoma. Radioiodine treatment planning for these patients is usually performed following the Benua-Leeper method, which constrains the administered activity to 2.96 GBq (80 mCi) whole-body retention at 48 h after administration to prevent lung toxicity in the presence of iodine-avid lung metastases. This limit was derived from clinical experience, and a dosimetric analysis of lung and tumor absorbed dose would be useful to understand the implications of this limit on toxicity and tumor control. Because of highly nonuniform lung density and composition as well as the nonuniform activity distribution when the lungs contain tumor nodules, Monte Carlo dosimetry is required to estimate tumor and normal lung absorbed dose. Reassessment of this toxicity limit is also appropriate in light of the contemporary use of recombinant thyrotropin (thyroid stimulating hormone) (rTSH) to prepare patients for radioiodine therapy. In this work we demonstrated the use of MCNP, a Monte Carlo electron and photon transport code, in a 3-dimensional (3D) imaging-based absorbed dose calculation for tumor and normal lungs. METHODS: A pediatric thyroid cancer patient with diffuse lung metastases was administered 37 MBq of (131)I after preparation with rTSH. SPECT/CT scans were performed over the chest at 27, 74, and 147 h after tracer administration. The time-activity curve for (131)I in the lungs was derived from the whole-body planar imaging and compared with that obtained from the quantitative SPECT methods. Reconstructed and coregistered SPECT/CT images were converted into 3D density and activity probability maps suitable for MCNP4b input. Absorbed dose maps were calculated using electron and photon transport in MCNP4b. Administered activity was estimated on the basis of the maximum tolerated dose (MTD) of 27.25 Gy to the normal lungs. Computational efficiency of the MCNP4b code was studied with a simple segmentation approach. In addition, the Benua-Leeper method was used to estimate the recommended administered activity. The standard dosing plan was modified to account for the weight of this pediatric patient, where the 2.96-GBq (80 mCi) whole-body retention was scaled to 2.44 GBq (66 mCi) to give the same dose rate of 43.6 rad/h in the lungs at 48 h. RESULTS: Using the MCNP4b code, both the spatial dose distribution and a dose-volume histogram were obtained for the lungs. An administered activity of 1.72 GBq (46.4 mCi) delivered the putative MTD of 27.25 Gy to the lungs with a tumor absorbed dose of 63.7 Gy. Directly applying the Benua-Leeper method, an administered activity of 3.89 GBq (105.0 mCi) was obtained, resulting in tumor and lung absorbed doses of 144.2 and 61.6 Gy, respectively, when the MCNP-based dosimetry was applied. The voxel-by-voxel calculation time of 4,642.3 h for photon transport was reduced to 16.8 h when the activity maps were segmented into 20 regions. CONCLUSION: MCNP4b-based, patient-specific 3D dosimetry is feasible and important in the dosimetry of thyroid cancer patients with avid lung metastases that exhibit prolonged retention in the lungs. PMID- 17138743 TI - Favorable biokinetic and tumor-targeting properties of 99mTc-labeled glucosamino RGD and effect of paclitaxel therapy. AB - Compared with the recent advancements in radiohalogenated Arg-Gly-Asp (RGD) peptides for alpha(v)beta(3)-targeted imaging, there has been limited success with (99m)Tc-labeled RGD compounds. In this article, we describe the favorable in vivo kinetics and tumor-imaging properties of a novel (99m)Tc-RGD compound that contains a glucosamine moiety. METHODS: Glucosamino (99m)Tc-d-c(RGDfK) was prepared by incorporating (99m)Tc(CO)(3) to the glucosamino peptide precursor in high radiochemical yield. Cell-binding characteristics were tested on human endothelial cells. Mice bearing RR1022 fibrosarcoma and Lewis lung carcinoma (LLC) tumors were used for in vivo biodistribution and blocking experiments and for imaging studies. Separate LLC-bearing mice underwent antiangiogenic therapy with 0, 20, or 40 mg of paclitaxel per kilogram of body weight every 2 d. Tumor volume was serially monitored, and tumor glucosamino (99m)Tc-d-c(RGDfK) uptake and Western blots of alpha(v) integrin expression were analyzed at day 14. RESULTS: Glucosamino (99m)Tc-d-c(RGDfK) binding to endothelial cells was dose dependently inhibited by excess RGD. Biodistribution in mice showed rapid blood clearance of glucosamino (99m)Tc-d-c(RGDfK), with substantially lower liver uptake and higher tumor uptake compared with (125)I-c(RGD(I)yV). Tumor uptake was 1.03 +/- 0.21 and 1.18 +/- 0.26 %ID/g at 1 h and 0.85 +/- 0.05 and 0.89 +/- 0.28 %ID/g at 4 h for sarcomas and carcinomas, respectively. Excess RGD blocked uptake by 76.5% and 70.2% for the respective tumors. gamma-Camera imaging allowed clear tumor visualization, with an increase of sarcoma-to-thigh count ratios from 5.5 +/- 0.7 at 1 h to 10.1 +/- 2.2 at 4 h and sustained carcinoma-to-thigh count ratios from 4 to 17 h. Pretreatment with excess cRGDyV significantly reduced tumor contrast on images. Paclitaxel therapy in LLC tumor-bearing mice significantly retarded tumor growth. This was accompanied by a corresponding reduction of tumor glucosamino (99m)Tc-d-c(RGDfK) uptake, which correlated significantly with tumor alpha(v) integrin expression levels. CONCLUSION: Glucosamino (99m)Tc-d-c(RGDfK) has favorable in vivo biokinetics and tumor imaging properties and may be useful for noninvasive evaluation of tumor integrin expression and response to antiangiogenic therapeutics. Because of the wide accessibility of gamma-cameras and high availability and excellent imaging characteristics of (99m)Tc, glucosamino (99m)Tc-d-c(RGDfK) may be an attractive alternative to radiohalogenated RGD peptides for angiogenesis-imaging research. PMID- 17138744 TI - Detection of alpha2-adrenergic receptors in brain of living pig with 11C yohimbine. AB - There have been few radiotracers for imaging adrenergic receptors in brain by PET, but none has advanced for use in human studies. We developed a radiosynthesis for the alpha(2)-adrenergic antagonist (11)C-yohimbine and characterized its binding in living pigs. As a prelude to human studies with (11)C-yohimbine, we determined the whole-body distribution of (11)C-yohimbine and calculated its dosimetry. METHODS: Yorkshire x Landrace pigs weighing 35-40 kg were used in the study. Baseline and postchallenge PET recordings of (11)C yohimbine in pig brain were conducted for 90 min, concurrent with arterial blood sampling, and with yohimbine and RX821002 as pharmacologic interventions. (15)O Water scans were performed to detect changes in cerebral perfusion. The PET images were manually coregistered to an MR atlas of the pig brain. Maps of the (11)C-yohimbine distribution volume ([V(d)] mL g(-1)) in brain were calculated relative to the arterial input function. RESULTS: Whole-body scans with (11)C yohimbine revealed high accumulation of radioactivity in kidney, intestine, liver, and bone. The estimated human dose was 5.6 mSv/GBq, a level commonly accepted in human PET studies. Brain imaging showed baseline values of V(d) ranging from 1.9 in medulla, 3.0 in cerebellum, and to 4.0 in frontal cortex. Coinjection with nonradioactive yohimbine (0.07 mg/kg) reduced V(d) globally to approximately 1.5-2 mL g(-1). A higher yohimbine dose (1.6 mg/kg) was without further effect on self-displacement. Very similar results were obtained by displacement with the more selective alpha(2)-adrenergic antagonist RX821002 at doses of 0.15 and 0.7 mg/kg. Cerebral blood flow was globally increased 43% after administration of RX821002. Notable features of (11)C-yohimbine are a lack of plasma metabolism over 90 min and a rapid approach to equilibrium binding in brain. CONCLUSION: The new radiotracer (11)C-yohimbine seems well suited for PET investigations of alpha(2)-adrenergic receptors in brain and peripheral structures, with the caveat that displaceable binding was present in cerebellum and throughout the brain. PMID- 17138745 TI - Radioimaging of light chain amyloid with a fibril-reactive monoclonal antibody. AB - Currently, there are no available means in the United States to document objectively the location and extent of amyloid deposits in patients with systemic forms of amyloidosis. To address this limitation, we have developed a novel diagnostic strategy, namely, the use of a radiolabeled fibril-reactive murine monoclonal antibody (mAb) as an amyloid-specific imaging agent. The goal of this study was to determine the pharmacokinetics, biodistribution, and ability of this reagent to target the type of amyloid that is formed from immunoglobulin light chains, that is, AL. METHODS: Subcutaneous tumors (amyloidomas) were induced in BALB/c mice by injection of human AL fibrils. The IgG1 mAb designated 11-1F4 and an isotype-matched control antibody were radioiodinated, and the pharmacokinetics and localization of these reagents were determined from blood and tissue samples. Amyloidoma-bearing animals that received (125)I- or (124)I-labeled antibodies were imaged by whole-body small-animal SPECT/CT or small-animal PET/CT technology, respectively. RESULTS: Radioiodinated mAb 11-1F4 retained immunoreactivity, as evidenced by its subnanomolar affinity for light chains immobilized on 96-well microtiter plates and for beads conjugated with a light chain-related peptide. Additionally, after intravenous administration, the labeled reagents had the expected biologic half-life of murine IgG1, with monoexponential whole-body clearance kinetics. In the amyloidoma mouse model, (125)I-11-1F4 was predominately localized in the tumors, as demonstrated in biodistribution and autoradiographic analyses. The mean uptake of this reagent, that is, the percentage injected dose per gram of tissue, 72 h after injection was significantly higher for amyloid than for skeletal muscle, spleen, kidney, heart, liver, or other tissue samples. Notably, the accumulation within the amyloidomas of (125)I- or (124)I-11-1F4 was readily visible in the fused small animal SPECT/CT or small-animal PET/CT images, respectively. CONCLUSION: Our studies demonstrate the amyloid-imaging capability of a radiolabeled fibril reactive mAb and provide the basis for a clinical trial designed to determine its diagnostic potential in patients with AL amyloidosis and other systemic amyloidoses. PMID- 17138746 TI - [Lys40(Ahx-DTPA-111In)NH2]exendin-4, a very promising ligand for glucagon-like peptide-1 (GLP-1) receptor targeting. AB - High levels of glucagon-like peptide-1 (GLP-1) receptor expression in human insulinomas and gastrinomas provide an attractive target for imaging, therapy, and intraoperative tumor localization, using receptor-avid radioligands. The goal of this study was to establish a tumor model for GLP-1 receptor targeting and to use a newly designed exendin-4-DTPA (DTPA is diethylenetriaminepentaacetic acid) conjugate for GLP-1 receptor targeting. METHODS: Exendin-4 was modified C terminally with Lys(40)-NH(2), whereby the lysine side chain was conjugated with Ahx-DTPA (Ahx is aminohexanoic acid). The GLP-1 receptor affinity (50% inhibitory concentration [IC(50)] value) of [Lys(40)(Ahx-DTPA)NH(2)]exendin-4 as well as the GLP-1 receptor density in tumors and different organs of Rip1Tag2 mice were determined. Rip1Tag2 mice are transgenic mice that develop insulinomas in a well defined multistage tumorigenesis pathway. This animal model was used for biodistribution studies, pinhole SPECT/MRI, and SPECT/CT. Peptide stability, internalization, and efflux studies were performed in cultured beta-tumor cells established from tumors of Rip1Tag2 mice. RESULTS: The GLP-1 receptor affinity of [Lys(40)(Ahx-DTPA)NH(2)]exendin-4 was found to be 2.1 +/- 1.1 nmol/L (mean +/- SEM). Because the GLP-1 receptor density in tumors of Rip1Tag2 mice was very high, a remarkably high tumor uptake of 287 +/- 62 %IA/g (% injected activity per gram tissue) was found 4 h after injection. This resulted in excellent tumor visualization by pinhole SPECT/MRI and SPECT/CT. In accordance with in vitro data, [Lys(40)(Ahx-DTPA-(111)In)NH(2)]exendin-4 uptake in Rip1Tag2 mice was also found in nonneoplastic tissues such as pancreas and lung. However, lung and pancreas uptake was distinctly lower compared with that of tumors, resulting in a tumor-to-pancreas ratio of 13.6 and in a tumor-to-lung ratio of 4.4 at 4 h after injection. Furthermore, in vitro studies in cultured beta-tumor cells demonstrated a specific internalization of [Lys(40)(Ahx-DTPA (111)In)NH(2)]exendin-4, whereas peptide stability studies indicated a high metabolic stability of the radiopeptide in beta-tumor cells and human blood serum. CONCLUSION: The high density of GLP-1 receptors in insulinomas as well as the high specific uptake of [Lys(40)(Ahx-DTPA-(111)In)NH(2)]exendin-4 in the tumor of Rip1Tag2 mice indicate that targeting of GLP-1 receptors in insulinomas may become a useful imaging method to localize insulinomas in patients, either preoperatively or intraoperatively. In addition, Rip1Tag2 transgenic mice represent a suitable animal tumor model for GLP-1 receptor targeting. PMID- 17138747 TI - 64Cu-azabicyclo[3.2.2]nonane thiosemicarbazone complexes: radiopharmaceuticals for PET of topoisomerase II expression in tumors. AB - Topoisomerase II (Topo-II) is an essential enzyme in the DNA replication process and is the primary cellular target for many of the most widely used and effective anticancer agents. It has been reported that thiosemicarbazones (TSCs) are potent antitumor agents that inhibit Topo-II. The aim of this study was to investigate the relationship between the in vitro and in vivo behavior of novel (64)Cu-TSC complexes and the expression of Topo-II activity. METHODS: Four (4)N azabicyclo[3.2.2]nonane TSC derivatives (EPH142, EPH143, EPH144, and EPH270) were successfully radiolabeled with (64)Cu, to form lipophilic cations of the general formula [(64)Cu(L)]Cl, and the partition coefficient (logP) values were determined. One agent [(64)Cu-EPH270](+) was observed in vitro in cultured cell studies. The kinetics of 2 compounds, [(64)Cu-EPH144](+) and [(64)Cu-EPH270](+), were examined in mice bearing L1210 tumors and small-animal PET was conducted in mice bearing L1210 and PC-3 tumors, which expressed high and low levels of Topo II, respectively. All data were compared with the activity and levels of Topo-II, as determined by a commercially available assay kit and western blot analysis. RESULTS: The 4 complexes were radiolabeled by incubation of (64)CuCl(2) with the ligand in ethanolic solution. The complexes were isolated in high radiochemical purity, as determined by radio-thin-layer chromatography and radio-high performance liquid chromatography. The compounds were shown to be lipophilic with logP values ranging from 1.34 to 1.92. In biodistribution studies, good L1210 tumor uptake was noted ([(64)Cu-EPH144](+) at 1 h, 4.70 %ID/g [percentage injected dose per gram]; 4 h, 8.80 %ID/g; 24 h, 6.64 %ID/g; and [(64)Cu EPH270](+) at 1 h, 2.58 %ID/g; 4 h, 6.00 %ID/g; 24 h, 4.80 %ID/g). Small-animal PET of animals with L1210 tumors (high Topo-II expressing) showed excellent tumor accumulation compared with that of animals with PC-3 tumors (low Topo-II expressing), and the L1210 tumor uptake was significantly reduced by coadministration of a Topo-II poison. CONCLUSION: Here we describe the characterization of a new class of copper-radiolabeled TSC analogs. We demonstrate that the accumulation of the (64)Cu-compounds is related to the expression levels of Topo-II in tumor tissue. PMID- 17138748 TI - Development of a novel 99mTc-chelate-conjugated bisphosphonate with high affinity for bone as a bone scintigraphic agent. AB - In bone scintigraphy using (99m)Tc with methylenediphosphonate ((99m)Tc-MDP) and hydroxymethylenediphosphonate ((99m)Tc-HMDP), it takes 2-6 h after an injection before imaging can start. This interval could be shortened with a new radiopharmaceutical with higher affinity for bone. Here, based on the concept of bifunctional radiopharmaceuticals, we designed a (99m)Tc mercaptoacetylglycylglycylglycine (MAG3)-conjugated hydroxy-bisphosphonate (HBP) ((99m)Tc-MAG3-HBP) and a (99m)Tc-6-hydrazinopyridine-3-carboxylic acid (HYNIC) conjugated hydroxy-bisphosphonate ((99m)Tc-HYNIC-HBP). METHODS: (99m)Tc-MAG3-HBP was prepared by complexation of MAG3-HBP with (99m)Tc using SnCl(2) as a reductant. The precursor of (99m)Tc-HYNIC-HBP, HYNIC-HBP, was obtained by deprotection of the Boc group after the coupling of Boc-HYNIC to a bisphosphonate derivative. (99m)Tc-HYNIC-HBP was prepared by a 1-pot reaction of HYNIC-HBP with (99m)TcO(4)(-), tricine, and 3-acetylpyridine in the presence of SnCl(2). Affinity for bone was evaluated in vitro by hydroxyapatite-binding assays for (99m)Tc-HMDP, (99m)Tc-MAG3-HBP, and (99m)Tc-HYNIC-HBP. Biodistribution experiments for the 3 (99m)Tc-labeled compounds were performed on normal rats. RESULTS: (99m)Tc-MAG3-HBP and (99m)Tc-HYNIC-HBP were each prepared with a radiochemical purity of >95%. In the in vitro binding assay, (99m)Tc-MAG3-HBP and (99m)Tc-HYNIC-HBP had greater affinity for hydroxyapatite than (99m)Tc-HMDP. In the biodistribution experiments, (99m)Tc-MAG3-HBP and (99m)Tc-HYNIC-HBP had higher levels of radioactivity in bone than (99m)Tc-HMDP. (99m)Tc-MAG3-HBP was cleared from the blood slower than (99m)Tc-HMDP, whereas there was no significant difference in clearance between (99m)Tc-HYNIC-HBP and (99m)Tc-HMDP. Consequently, (99m)Tc-HYNIC-HBP showed a higher bone-to-blood ratio than (99m)Tc-HMDP. CONCLUSION: We developed a novel (99m)Tc-chelate-conjugated bisphosphonate with high affinity for bone and rapid clearance from blood, based on the concept of bifunctional radiopharmaceuticals. The present findings indicate that (99m)Tc HYNIC-HBP holds great potential for bone scintigraphy. PMID- 17138749 TI - PET of vascular endothelial growth factor receptor expression. AB - For solid tumors and metastatic lesions, tumor vascularity is a critical factor in assessing response to therapy. Here we report the first example, to our knowledge, of (64)Cu-labeled vascular endothelial growth factor 121 (VEGF(121)) for PET of VEGF receptor (VEGFR) expression in vivo. METHODS: VEGF(121) was conjugated with 1,4,7,10-tetraazadodecane-N,N',N'',N'''-tetraacetic acid (DOTA) and then labeled with (64)Cu for small-animal PET of mice bearing different sized U87MG human glioblastoma xenografts. Blocking experiments and ex vivo histopathology were performed to confirm the in vivo results. RESULTS: There were 4.3 +/- 0.2 DOTA molecules per VEGF(121), and the VEGFR2 binding affinity of DOTA VEGF(121) was comparable to VEGF(121). (64)Cu labeling of DOTA-VEGF(121) was achieved in 90 +/- 10 min and the radiolabeling yield was 87.4% +/- 3.2%. The specific activity of (64)Cu-DOTA-VEGF(121) was 3.2 +/- 0.1 GBq/mg with a radiochemical purity of >98%. Small-animal PET revealed rapid, specific, and prominent uptake of (64)Cu-DOTA-VEGF(121) in small U87MG tumors (high VEGFR2 expression) but significantly lower and sporadic uptake in large U87MG tumors (low VEGFR2 expression). No appreciable renal clearance of (64)Cu-DOTA-VEGF(121) was observed, although the kidney uptake was relatively high likely due to VEGFR1 expression. Blocking experiments, immunofluorescence staining, and western blot confirmed the VEGFR specificity of (64)Cu-DOTA-VEGF(121). CONCLUSION: Successful demonstration of the ability of (64)Cu-DOTA-VEGF(121) to visualize VEGFR expression in vivo may allow for clinical translation of this radiopharmaceutical for imaging tumor angiogenesis and guiding antiangiogenic treatment, especially patient selection and treatment monitoring of VEGFR-targeted cancer therapy. PMID- 17138750 TI - Effects of antifolate drugs on the cellular uptake of radiofolates in vitro and in vivo. AB - Targeting the folate receptor (alpha-FR) with radiolabeled folates for the noninvasive diagnosis and therapy of alpha-FR-overexpressing neoplastic tissue is of great interest. However, the tumor uptake of folate-based radiotracers was shown to be low compared with the high renal retention of radioactivity attributable to alpha-FR expression in the proximal tubule cells. In order to increase the tumor uptake of radiofolates, we wanted to stimulate alpha-FR expression or transport through coapplication of the antifolates methotrexate (MTX), raltitrexed (RTX), and pemetrexed (PMX). METHODS: (99m)Tc-picolylamine monoacetic acid folate ((99m)Tc-PAMA-folate) was used for these studies. The in vitro experiments with antifolates were performed with alpha-FR-positive KB cancer cells. In vivo experiments were performed with KB tumor-bearing athymic nude mice. In vivo images were acquired with a small-animal SPECT/CT scanner. RESULTS: KB cells incubated with solutions (10 micro mol/L) of MTX, RTX, or PMX for 24 h displayed twice as much (99m)Tc-PAMA-folate uptake as untreated cells. In contrast, KB tumor-bearing mice that received MTX intravenously 24 h before (99m)Tc-PAMA-folate showed significantly lower uptake of the radiofolate in tumors (1.35 +/- 0.33 percentage injected dose per gram of tissue [%ID/g] [mean +/- SD]) and the alpha-FR-positive kidneys (9.35 +/- 1.73 %ID/g) than did control mice (2.33 +/- 0.36 and 18.48 +/- 0.72 %ID/g, respectively, at 4 h after injection). When the antifolate PMX and (99m)Tc-PAMA-folate were injected 1 h apart, the tumor uptake of the radiotracer was unaffected (2.21 +/- 0.34 %ID/g at 4 h after injection), whereas radioactivity in the kidneys was significantly decreased (1.14 +/- 0.18 %ID/g at 4 h after injection). In vivo SPECT/CT studies demonstrated the specific accumulation of (99m)Tc-PAMA-folate in tumors and almost a complete absence of radioactivity in the renal tissue of mice preinjected with PMX. CONCLUSION: Our data suggest that the preadministration of antifolates improves tumor-to-kidney ratios of radiofolates and opens a "therapeutic window" for folates radiolabeled with particle-emitting nuclides, which could otherwise be nephrotoxic. PMID- 17138751 TI - A picture is worth. PMID- 17138752 TI - The role of modern biology and medicine in drug development in academia and industry. AB - This symposium addresses careers in drug development in industry; the performance of translational research by academia, industry, and both; and numerous factors pertinent to alliances essential to drug discovery and development. Drug development is a complex process that regularly involves effective collaborations between academic and physician scientists and industry. There are specific occupational factors affecting recruitment of scientists and physicians in drug development programs in industry; ideal backgrounds for successful applicants for positions in industry in drug development; ethical and regulatory considerations particularly germane to the performance of scientists and physicians in drug development programs in industry and at universities; and particular gratifications available to scientists in industry working on drug development. Both similarities and differences characterize the performance of translational research in industry compared with academia. In industry, logistic, operational, and scientific oversight is complex, especially because it often involves relationships with clinical enterprises outside of the corporation. The process is long and arduous from formulation of a good idea in discovery to acceptance of a novel drug in the marketplace. Collaborations and partnerships by industry often involving academia and confrontation of multiple issues are pivotal. PMID- 17138753 TI - Transition from academia to industry: a personal account. AB - A career in industry has become a widely accepted alternative for those of us trained in medicine and/or science who have traditionally focused on careers in academia. Like any career decision, consideration of a position in industry should include asking yourself a series of fundamental questions. A few of the key questions should include: 1) What kind of work environment do you find most enjoyable? (e.g., patient care setting, basic research lab, team-oriented setting); 2) What are you focused on accomplishing in your career? (basic research discoveries, contributions to clinical medicine, compensation); 3) Are you team oriented in your interactions or are you more of an individual contributor? A successful career in any endeavor, including industry, starts with a careful and honest examination of what you are best suited for and inspired to do. PMID- 17138754 TI - Translational research in academia and industry. AB - Research that bridges between scientific insights and clinical application is one of the most active and exciting areas of current biomedical activity. Much of this translational work occurs through collaborations between academic and industrial institutions, taking advantage of the respective strengths and resources of the two sectors. However, such collaborations sometimes can be challenging due to differences between the cultures and priorities of the two parties. This article discusses the nature of translational research, with a focus on the academia-industry interface, analyzes the factors important for effective collaborations, and describes specific examples of successful translational research programs. PMID- 17138755 TI - Effective partnering of academic and physician scientists with the pharmaceutical drug development industry. AB - This manuscript briefly addresses the drug discovery and development process. It is a long road from the formulation of a good discovery idea to the acceptance of a new drug in the marketplace, and there are many challenges faced along the way to the patient. Collaborations and partnerships are an important part of this process. There are a variety of partnering opportunities, ranging from the discovery of novel technologies and drug targets to lead discovery, compound gifts, and external sourcing. These partnerships help increase confidence and improve decision making on issues of safety and efficacy preclinically, which can reduce attrition and expedite the provision of new quality drugs to patients more quickly and at lower costs. Collaborations involve addressing multiple issues that include infrastructure, safety, regulatory matters, intellectual property, technical and personnel considerations, source document capture and data analysis issues, and legal and strategic alliances. A number of success factors are identified as important for quality collaborations in the drug development process. PMID- 17138756 TI - Gut responses to enteral nutrition in preterm infants and animals. AB - Preterm birth is associated with immature digestive function that may require the use of total parenteral nutrition and special oral feeding regimens. Little is known about the responses to oral food in the preterm neonate and how enteral nutrients affect the immature gastrointestinal tract (GIT). In vivo studies are difficult to perform in laboratory rodents because of their small body size and that of immature organs at birth, and this makes the large farm animals (e.g., pigs, cattle, sheep) more attractive models in this field. In these species, preterm delivery at 88%-95% gestation is associated clinical complications and degrees of GIT immaturity similar to those in infants born at 70%-90% gestation. Studies in both animals and infants indicate that the immature GIT responds to the first enteral food with rapid increases in gut mass and surface area, blood flow, motility, digestive capacity, and nutrient absorption. To a large extent, the enteral food responses are birth independent, and can be elicited also in utero, at least during late gestation. Nevertheless, preterm neonates show compromised GIT structure, function, and immunology, particularly when delivered by caesarean section and fed diets other than mother's milk. Formula-fed preterm infants are thus at increased risk of developing diseases such as necrotizing enterocolitis, unless special care is taken to avoid excessive nutrient fermentation and bacterial overgrowth. The extent to which results obtained in preterm animals (most notably the pig) can be used to reflect similar conditions in preterm infants is discussed. PMID- 17138757 TI - In vitro evaluation for potential calcification of biomaterials used for staple line reinforcement in lung surgery. AB - Bovine pericardium (BPC) and polytetrafluoroethylene (PTFE) have been widely used to reinforce staple lines in lung resection. Since limited information regarding the calcification of these biomaterials is available, we undertook an in vitro study to evaluate their calcification potential. Commercially available BPC and PTFE biomaterials were evaluated and compared with custom-prepared BPC tissue. In vitro calcification was performed via submersion in supersaturated solution in a double-walled glass reactor at 37.0 degrees C +/- 0.1 degrees C, pH 7.4 +/- 0.1, mimicking most ion concentrations of human blood plasma. In processing of calcification, the pH decrease of the solution simulated the addition of consumed H(+), Ca(2+), and PO(4)(3-) ions from titrant solutions, the concentrations of which were based on the stoichiometry of octacalcium phosphate. The molar ion addition with time was recorded, and the initial slope of the curve was computed for each experiment. The rate of calcification developed (molar calcium phosphate ion addition rate per time and total surface area) (R) was computed after that with respect to the relative supersaturation (sigma) used in each experiment. R for custom-prepared BPC tissues was found to be in the range of 0.19 +/- 0.08 to 0.52 +/- 0.19 (n = 17) in sigma range of 0.72 to 1.42. Commercial BPC was found to be 0.016 to 0.052 (n = 4), and PTFE was 0.005 to 0.05 (n = 8) in the same sigma range. Both clinically applied biomaterials, BPC and PTFE, seemed to be calcified with rates of at least one order of magnitude lower than the custom prepared BPC tissue. This data suggested that BPC and PTFE biomaterials showed a similar, relatively very low tendency for calcification compared with custom prepared BPC tissue. Although further studies are necessary, staple line reinforcement by these two biomaterials should be considered safe from the calcification point of view. PMID- 17138758 TI - Characterization of ovarian surface epithelial cells from the hen: a unique model for ovarian cancer. AB - To further develop the hen as a model of ovarian adenocarcinoma, we have studied normal and neoplastic ovaries as well as cultured cells from the ovarian surface epithelium (OSE). We characterized the OSE layer of the hen for specific histologic markers and evaluated these markers on tumor tissue. We also isolated and characterized the epithelial cells that are the likely source of the ovarian tumors of the hen. The surface epithelium of normal ovaries demonstrated positive staining for cytokeratin, proliferating cell nuclear antigen (PCNA), progesterone receptor (PR), and negative staining for vimentin. Ovarian tumors demonstrated positive cytokeratin, PCNA, PR, and weak vimentin staining in the gland-like areas. Epithelial cell cultures were obtained by an explant method utilizing small and large yellow follicles. These cells were positive for cytokeratin and negative for vimentin on Days 1 and 3. By Day 10, cytokeratin protein expression was less for some cells, and vimentin expression was weakly present in some cells. Expression of PCNA was observed at Days 1 and 3, but was rarely seen in cells cultured for 10 days. Expression of PR was observed on Day 10 after 24-hr estrogen treatment. Epithelial cells grew slowly in culture, and were susceptible to trypsin or other dissociation treatments. PMID- 17138759 TI - Impact of Cu, Zn-superoxide dismutase and Se-dependent glutathione peroxidase-1 knockouts on acetaminophen-induced cell death and related signaling in murine liver. AB - There is increasing evidence showing dual functions of antioxidant enzymes in coping with reactive oxygen species (ROS) versus reactive nitrogen species (RNS). The objective of this study was to compare the impacts of knockout of Cu, Zn superoxide dismutase (SOD1) and Se-dependent glutathione peroxidase-1 (GPX1) on cell death and related signaling mediated by acetaminophen (APAP), a RNS inducer in liver. Two groups of young adult knockout mice (SOD1(-/-) and GPX1(-/-)), along with their wild types (WT), were killed 5 hrs after an ip injection of saline or APAP (300 mg/kg body wt). While the WT mice showed more hepatic necrosis and DNA breakage than the GPX1(-/-) mice, the SOD1(-/-) mice had essentially no positive response compared with their saline-injected controls. The APAP treatment activated liver c-jun N-terminal kinase (JNK) in the WT and GPX1(-/-) mice, but not in the SOD1(-/-) mice. The APAP-induced changes in other cell death-related signal proteins such as p21, caspase-3, and poly(ADP-ribose) polymerase (PARP) also were obviated in the SOD1(-/-) mice. In conclusion, knockout of GPX1 did not potentiate APAP-induced cell death and related signaling, whereas the SOD1 null blocked APAP-induced hepatic JNK phosphorylation and cell death. PMID- 17138760 TI - Bioavailability of carotenoids and tocopherols from broccoli: in vivo and in vitro assessment. AB - Fruits and vegetables are the major sources of biologically active compounds, and carotenoids and tocopherols constitute important groups in human diets. Bioavailability is a critical feature in the assessment of the role of micronutrients in human health, and the approaches to this issue include in vitro and in vivo methods. Our aim was to evaluate the bioavailability of carotenoids and tocopherols present in broccoli and to compare in vitro and in vivo approaches. Fourteen apparently healthy volunteers consumed 200 g broccoli once a day for seven days. Blood samples were drawn at baseline and after intervention to determine changes in lutein, beta-carotene, and alpha- and gamma-tocopherol as relevant phytochemicals provided with this vegetable. Broccoli also was subjected to simulated gastrointestinal digestion to assess changes related to preabsorptive processes. Analytes in serum and at each phase of the digestion were assayed by high-performance liquid chromatography. During the intervention, the amounts supplied daily ranged from 2.4 to 3.1 mg lutein, 1.4 to 1.8 mg beta carotene, 4.5 to 6.8 mg alpha-tocopherol, and 0.8 to 1.8 mg gamma-tocopherol. Significant changes in serum in both men and women were observed only for lutein, whereas for gamma-tocopherol a significant change was detected in women. No changes were observed for alpha-tocopherol, beta-carotene, retinol, the alpha tocopherol-to-cholesterol ratio, or serum lipids. Using the in vitro model, more than 75% of lutein, beta-carotene, gamma-tocopherol, and alpha-tocopherol remained at the duodenal phase, whereas incorporation into the supernatants accounted for <20% of the initial content in food. Regular consumption of broccoli at dietary levels increased serum concentrations of lutein and gamma tocopherol without affecting alpha-tocopherol or beta-carotene status in serum. The behavior of these phytochemicals under in vitro gastrointestinal conditions does not fully explain the changes observed in vivo. PMID- 17138761 TI - Hepatocytes detoxify Atuna racemosa extract. AB - A number of traditional medicine plants are hepatotoxic. Thus, while the traditional uses of Atuna racemosa suggest little indication for toxicity, it is nonetheless important to examine the potential for this extract to target the liver. Using Jurkat T cells and HepG2 hepatocytes as a model, the potential hepatotoxicity of this extract was evaluated. The results of a conditioned media experiment suggest that A. racemosa extract would likely be detoxified by the liver. These results provide the necessary background to initiate an in vivo toxicology investigation. PMID- 17138762 TI - Epoxyeicosatrienoic Acid inhibition alters renal hemodynamics during pregnancy. AB - In this study we examined the expression of cytochrome P450 (CYP) 2C and CYP2J isoforms in renal proximal tubules and microvessels isolated from rats at different stages of pregnancy. We also selectively inhibited epoxyeicosatrienoic acid (EET) production by the administration of N-methanesulfonyl-6-(2 proparyloxyphenyl)hexanamide (MSPPOH 20 mg/kg/day iv) to rats during Days 14-17 of gestation and to age-matched virgin rats and determined the consequent effects on renal function. Western blot analysis showed that CYP2C11, CYP2C23, and CYP2J2 expression was significantly increased in the renal microvessels of pregnant rats on Day 12 of gestation. In the proximal tubules, CYP2C23 expression was significantly increased throughout pregnancy, while the expression of CYP2C11 was increased in early and late pregnancy and the expression of CYP2J2 was increased in middle and late pregnancy. MSPPOH treatment significantly increased pregnant rats' mean arterial pressure, renal vascular resistance, and sodium balance but significantly decreased renal blood flow, glomerular filtration rate, and urinary sodium excretion, as well as fetal pups' body weight and length. In contrast, MSPPOH treatment had no effect on renal hemodynamics or urinary sodium excretion in age-matched virgin rats. In pregnant rats, MSPPOH treatment also caused selective inhibition of renal cortical EET production and significantly decreased the expression of CYP2C11, CYP2C23, and CYP2J2 in the renal cortex, renal microvessels, and proximal tubules. These results suggest that upregulation of renal vascular and tubular EETs contributes to the control of blood pressure and renal function during pregnancy. PMID- 17138763 TI - Molecular and pharmacological properties of human embryonic stem cell-derived cardiomyocytes. AB - Human embryonic stem cells (hESCs) can be coaxed to differentiate into specific cell types, including cardiomyocyte-like cells. These cells express cardiac specific markers and display functional similarities to their adult counterparts. Based on these properties, hESC-derived cardiomyocytes have the potential to be extremely useful in various in vitro applications and to provide the opportunity for cardiac cell replacement therapies. However, before this can become a reality, the molecular and functional characteristics of these cells need to be investigated in more detail. In the present study we differentiate hESCs into cardiomyocyte-like cells via embryoid bodies (EBs). The fraction of spontaneously beating clusters obtained from the EBs averaged approximately 30% of the total number of EBs used. These cell clusters were isolated, dissociated into single cell suspensions, and frozen for long-term storage. The cryopreserved cells could be successfully thawed and subcultured. Using electron microscopy, we observed Z discs and tight junctions in the hESC-derived cardiomyocytes, and by immunohistochemical analysis we detected expression of cardiac-specific markers (cTnI and cMHC). Notably, using BrdU labeling we also could demonstrate that some of the hESC-derived cardiomyocytes retain a proliferative capacity. Furthermore, pharmacological stimulation of the cells resulted in responses indicative of functional adrenergic and muscarinic receptor coupling systems. Taken together, these results lend support to the notion that hESCs can be used as a source for the procurement of cardiomyocytes for in vitro and in vivo applications. PMID- 17138765 TI - Hormones and breast cancer: should practice be changed? PMID- 17138766 TI - Breast cancer risk in postmenopausal women using estrogen-only therapy. AB - OBJECTIVE: To evaluate whether the risk of estrogen-only therapy on breast cancer varies by dose, constituent, and route of administration. METHODS: All Finnish women older than age 50 years using oral or transdermal estradiol (n=84,729), oral estriol (n=7,941), or vaginal estrogens (n=18,314) for at least 6 months during 1994-2001 were identified from the national medical reimbursement register. They were followed for breast cancer with the aid of the Finnish Cancer Registry to the end of 2002. RESULTS: Altogether, 2,171 women with breast cancer were identified. The standardized incidence ratio of breast cancer with systemic estradiol for less than 5 years was 0.93 (95% confidence interval 0.80-1.04), and for estradiol use for 5 years or more, 1.44 (1.29-1.59). Oral and transdermal estradiol was accompanied by a similar risk of breast cancer. The risk was most prominent with the dose greater than 1.9 mg/d orally; whereas the risk associated with transdermal route was not dose-dependent. The standardized incidence ratio for the lobular type of breast cancer (1.58) was slightly higher than that for the ductal type (1.36). The use of estradiol was associated with both localized breast cancer (1.45; 1.26-1.66) and cancer spread to regional nodes (1.35; 1.09 1.65). The incidence of carcinoma in situ (n=32) was increased (2.43; 1.66-3.42) among estradiol users. CONCLUSION: Estradiol for 5 years or more, either orally or transdermally, means 2-3 extra cases of breast cancer per 1,000 women who are followed for 10 years. Oral estradiol use for less than 5 years, oral estriol, or vaginal estrogens were not associated with a risk of breast cancer. LEVEL OF EVIDENCE: II-2. PMID- 17138767 TI - Effect of human papillomavirus vaccines on vulvar, vaginal, and anal intraepithelial lesions and vulvar cancer. AB - OBJECTIVE: Human papillomavirus (HPV) is a necessary cause for cervical cancer, and it has been associated with vulvar and vaginal cancer and vulvar (VIN) and vaginal (VaIN) and anal (AIN) intraepithelial neoplasia. We assessed the prevalence of HPV (and the types) to estimate the possible effect of a HPV vaccine on lower genital tract disease prevention. METHODS: Two hundred fifty eight samples of VIN, VaIN, AIN, and vulvar cancer from 241 women were included in the study. The diagnosis of surgical samples was made using published histomorphologic criteria. The DNA was extracted for HPV detection and typed using polymerase chain reaction and sequencing. RESULTS: The analyses were performed on 210 intraepithelial neoplasia samples (VIN2/3, VaIN2/3, AIN2/3) and 48 vulvar carcinoma samples. Human papillomavirus DNA was detected in 92%, 91%, 89%, and 60% of the VIN, VaIN, AIN, and vulvar carcinoma samples, respectively. High-risk HPV types 16 or 18 were detected in 76%, 64%, 81%, and 42% of the VIN2/3, VaIN2/3, AIN, and vulvar carcinoma samples. Women with HPV-positive samples were younger than those with HPV-negative samples (46 years compared with 55 years and 51 years compared with 61 years, for the VIN2/3 and vulvar carcinoma samples, respectively). Human papillomavirus-positive vulvar carcinoma was more frequent in women aged younger than 56 years (77%), than in those aged 56 years or older (41%). CONCLUSION: Based on the data obtained in this study, widely implemented prophylactic HPV vaccination could make an important contribution to the reduction of the risk for cervical cancer and could also prevent about half the vulvar carcinomas in younger women and about two thirds of the intraepithelial lesions in the lower genital tract. LEVEL OF EVIDENCE: II-3. PMID- 17138768 TI - Cervical intraepithelial neoplasia in adolescent women: incidence and treatment outcomes. AB - OBJECTIVE: We sought to estimate the incidence of cervical intraepithelial neoplasia (CIN) and treatment outcomes in adolescents with abnormal cytology. METHODS: Adolescent women (ages 14-21 years) referred to colposcopy clinic for abnormal cytology from 1992 to 2004 were identified by computerized database. Only adolescents with biopsy-proven CIN were evaluated. Demographic and risk factor data were obtained from medical records. Referral cytology, histology on biopsy and loop electrosurgical excisional procedure (LEEP), and follow-up cytology were analyzed and compared. Statistical analysis was performed by chi(2) or Fisher exact test, Student t tests, and logistic regression. RESULTS: Of 1,678 adolescents, 517 had biopsy-proven CIN and follow-up. Seventy-seven patients were referred with atypical squamous cells of undetermined significance (ASCUS) cytology; 174 patients were referred with low-grade squamous intraepithelial lesions (LSIL), 258 with high-grade squamous intraepithelial lesions (HSIL) and eight with atypical glandular cells (AGC). The rate of CIN 2/3 in patients with ASCUS, LSIL, and HSIL was 35% (95% confidence interval 24-46%), 36% (29-43%), and 50% (44-56%), respectively. A total of 192 patients with biopsy-proven CIN 2/3 underwent a LEEP. No patients were diagnosed with cervical carcinoma. Fifty-five percent (95% confidence interval 48-62%) of patients had abnormal cytology on follow-up, suggesting recurrence or reinfection. CONCLUSION: Adolescents with abnormal cytology have a high incidence of CIN2/3 and high rates of abnormal cytology after LEEP. Cervical intraepithelial neoplasia 2/3 is common in adolescents with abnormal cytology, yet no cases of cancer were identified. Importantly, LEEP fails to meet its therapeutic goals given a high incidence of abnormal follow-up cytology and may represent overly aggressive therapy because the majority of human papillomavirus infections are transient with high regression rates. LEVEL OF EVIDENCE: III. PMID- 17138769 TI - A prospective blinded evaluation of the accuracy of frozen section for the surgical management of endometrial cancer. AB - OBJECTIVE: To prospectively evaluate in a blinded fashion the accuracy of frozen section in endometrial cancer. METHODS: Sixty patients with endometrial cancer or complex atypical hyperplasia were consecutively enrolled. Intraoperatively, a frozen section was obtained, processed, and stored for interpretation by blinded pathologists. Final pathologic diagnosis was conducted in the usual fashion with the pathologists blinded to frozen results. Histologic grade and myometrial invasion on frozen section was correlated with final pathology. RESULTS: Median age was 61 years (range, 39-82 years). Fifty-seven percent of patients were white, and mean body mass index was 40 mg/kg2. Depth of invasion on frozen correlated with final pathology in 67% (95% confidence interval [CI] 55-79%). Twenty-eight percent (95% CI 17-39%) of patients were upstaged from frozen to final. Patients with no invasion on frozen were upstaged in 46% (95% CI 28-64%). Histologic grade on frozen correlated with final pathology in 58% (95% CI 46 70%); 38% (95% CI 26-50%) of patients were upgraded by final grade. Patients with frozen grade 1 histology or less were upgraded in 61% (95% CI 45-77%). Clinically relevant upstaging occurred in 11 patients (18%) (95% CI 8-28%). CONCLUSION: Frozen section for histologic grade and depth of myometrial invasion in endometrial cancer correlates poorly with final pathology. Because a large number of patients are potentially understaged with the use of frozen section with a subsequent risk of over and under treatment, we recommend consideration of comprehensive surgical staging for all patients with endometrial cancer. LEVEL OF EVIDENCE: II-2. PMID- 17138770 TI - Effect of mifepristone for symptomatic leiomyomata on quality of life and uterine size: a randomized controlled trial. AB - OBJECTIVE: To assess the effect of low-dose mifepristone on quality of life, pain, bleeding, and uterine size among women with symptomatic leiomyomata. METHODS: Forty-two women with symptomatic uterine leiomyomata and uterine volume of 160 mL or more were randomized to mifepristone, 5 mg daily, or placebo for 26 weeks. Quality of life (Uterine Fibroid Symptoms Quality of Life Questionnaire and Medical Outcomes Study 36-Item Short Form survey) and uterine and leiomyoma size (ultrasonography) were assessed at baseline, and at 1 month, 3 months, and 6 months of treatment. Bleeding (daily logs and pictorial charts) and pain (McGill Pain Questionnaire) were assessed monthly. Endometrial pathology was assessed at baseline and 6 months. RESULTS: Forty-two women were randomized; 37 women completed all 6 months. Women randomized to mifepristone showed an improvement in leiomyoma-specific quality of life. Forty-one percent became amenorrheic, rates of anemia improved, and adjusted uterine size was reduced by 47%. Compared with the placebo group, improvements in these outcomes in the treatment group were significantly greater (P<.05 to .001). There were no significant differences in adverse effects between the groups. No endometrial hyperplasia was noted in any participant. CONCLUSION: Low-dose mifepristone improves leiomyoma-specific quality of life and reduces leiomyoma size among women with symptomatic leiomyomata. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov www.clinicaltrials.gov NCT00133705 LEVEL OF EVIDENCE: I. PMID- 17138771 TI - Pelvic organ prolapse in nulliparous women and their parous sisters. AB - OBJECTIVE: To investigate the role of vaginal delivery and familial factors in the development of pelvic organ prolapse by comparing the prevalence of this condition in nulliparous women and their parous sisters. METHODS: Pairs of nulliparous and parous postmenopausal sisters were recruited for assessment of pelvic organ prolapse. One hundred one sister pairs underwent clinical evaluation. Pelvic organ prolapse was recorded using the Pelvic Organ Prolapse Quantification System. RESULTS: The majority of women had no pelvic organ prolapse. By compartment there was a 74.3% to 91.1% concordance in prolapse stage within sister pairs. In discordant sister pairs, the parous sister was found to have the more advanced prolapse 88% of the time. CONCLUSION: High concordance of pelvic organ prolapse in nulliparous and parous sister pairs suggests a familial predisposition toward developing this condition. However, vaginal delivery does appear to confer a risk for more advanced pelvic organ prolapse. LEVEL OF EVIDENCE: II-2. PMID- 17138772 TI - Endoanal ultrasound findings and fecal incontinence symptoms in women with and without recognized anal sphincter tears. AB - OBJECTIVE: To estimate whether endoanal ultrasound findings are more prevalent in primiparous women with a history of anal sphincter tear than in women without this history and whether the findings are associated with fecal incontinence symptoms. METHODS: A total of 251 primiparous women at seven clinical sites underwent standardized ultrasound assessment of the internal and external anal sphincter 6-12 months after delivery. Participants were women in the three cohorts of the Childbirth and Pelvic Symptoms Study: 1) women with clinically evident third- or fourth-degree tear at vaginal delivery (n=106); 2) no tear at vaginal delivery (n=106); and 3) cesarean delivery without labor (n=39). Women completed the Fecal Incontinence Severity Index to assess fecal incontinence symptoms. RESULTS: Thirty-five percent of the sphincter tear group exhibited internal sphincter gaps compared with 3% of vaginal controls (odds ratio [OR] 18.4, 95% confidence interval [CI] 5.5-62.1) and 10% of cesarean controls. External sphincter gaps were identified in 51% of the tear group compared with 31% of vaginal controls (OR 2.3, 95% CI 1.3-4.0) and 28% of cesarean controls. In the tear group, fecal incontinence severity was greater in those with internal sphincter gaps compared with those with no internal sphincter gaps (Fecal Incontinence Severity Index score 6.6+/-8.3 compared with 3.3+/-6.1, P=.02), as well as in those with external sphincter gaps (6.1+/-8.4 compared with 2.7+/-5.0, P=.01), and greatest in those with both internal and external sphincter gaps compared with at least one gap not present (7.2+/-8.1 compared with 3.4+/-6.4, P=.003). CONCLUSION: Anal sphincter gaps detected by ultrasonography are prevalent in postpartum primiparous women with a history of sphincter tear and are associated with fecal incontinence severity. LEVEL OF EVIDENCE: II-2. PMID- 17138773 TI - Eszopiclone in patients with insomnia during perimenopause and early postmenopause: a randomized controlled trial. AB - OBJECTIVE: To evaluate eszopiclone 3 mg for treatment of insomnia in perimenopausal and early postmenopausal women, as well as the impact of insomnia treatment on mood, menopause-related symptoms, and quality of life. METHODS: This was a double-blind, placebo-controlled study with 410 women (aged 40-60; perimenopausal or early postmenopausal) who reported insomnia defined as sleep latency of at least 45 minutes and total sleep time less than or equal to 6 hours per night for at least 3 nights per week over the previous month. Patients were randomly assigned to eszopiclone 3 mg or placebo nightly for 4 weeks. Sleep data were collected once a day. Physician global assessments of menopause, menopause specific questionnaire, Greene Climacteric Scale, the Montgomery Asberg Depression Rating Scale, and the Sheehan Disability Scale were collected at baseline and end of treatment. RESULTS: Patients receiving eszopiclone reported improvements in sleep induction, sleep maintenance, sleep duration, sleep quality, and next-day functioning relative to placebo (P<.05). Patients receiving eszopiclone reported fewer total awakenings and awakenings due to hot flushes (P<.05). Eszopiclone use led to greater improvement in Montgomery Asberg Depression Rating Scale scores (P<.05) and physician global assessments of menopause scores (P<.001); total Greene Climacteric Scale score and the vasomotor and psychological sub-scores (P<.05); vasomotor and physical domains of the menopause-specific questionnaire (P<.05); and family life/home domain of the Sheehan Disability Scale (P<.05). CONCLUSION: In this study, eszopiclone provided significant improvements in sleep and positively impacted mood, quality of life, and menopause-related symptoms in perimenopausal and early postmenopausal women with insomnia. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov www.clinicaltrials.gov NCT00366093 LEVEL OF EVIDENCE: I. PMID- 17138774 TI - Fear of inability to conceive in pregnant adolescents. AB - OBJECTIVE: To estimate the demographic and health history differences between pregnant adolescents who had fears that they would not be able to conceive and those without these fears. METHODS: Three hundred pregnant adolescents presenting for their first prenatal visit participated in a cohort study that addressed attitudes about pregnancy. All participants were aged younger than 20 years and gave informed consent. The outcome of interest was a positive response to the question "Did you have any fears that you wouldn't be able to get pregnant?" Independent measures included health history and demographic variables. RESULTS: Among participants, 42% stated they had fears about not being able to conceive. The total sample included 20% 12-15 year olds, 39% 16-17 year olds and 41% 18-19 year-olds. There was no statistically significant difference in fear of not being able to conceive by maternal age, reported sexually transmitted disease rates, or age at first intercourse. More adolescents who expressed fear of infertility had a previous spontaneous abortion, previous pelvic examination, and were sexually active for a longer period of time compared with those without this fear. CONCLUSION: A large proportion of pregnant adolescents in this study expressed fear that they would not be able to conceive. Understanding the basis of the fear is critical to appreciating its association with current and future adolescent pregnancy and contraceptive use. LEVEL OF EVIDENCE: III. PMID- 17138775 TI - Young pregnant women's knowledge of modern intrauterine devices. AB - OBJECTIVE: Modern intrauterine devices (IUDs) are safe, effective, and reversible, but only 2.1% of U.S. women use IUDs. We aimed to estimate young pregnant women's knowledge of IUDs. METHODS: We surveyed 190 women, aged 14-25 years, presenting for prenatal or abortion care about their contraceptive history, plans, and knowledge. We asked if they had heard of IUDs and queried them on IUD characteristics. RESULTS: The women were, on average, 20 years old, 27% had education past high school, and 47% had delivered a child. Half were in prenatal care, and 91% had not planned their current pregnancy. Fifty-two percent wished to wait 4 or more years before their next pregnancy, and 27% did not want to be pregnant ever again. Safety and efficacy were the most important factors in choosing a contraceptive method. Fifty percent had heard of IUDs, 71% did not know about IUDs' safety, and 58% did not know about IUDs' efficacy. Respondents who knew of IUDs were older (21 versus 19 years, P<.001) and more likely to be parous (55% versus 39%, P=.04). CONCLUSION: Young women choosing contraception after a pregnancy would benefit from counseling about the relative safety and effectiveness of IUDs, allowing them to make fully informed contraceptive decisions. LEVEL OF EVIDENCE: II-2. PMID- 17138776 TI - Effects of human immunodeficiency virus on protracted amenorrhea and ovarian dysfunction. AB - OBJECTIVE: To characterize ovarian failure and prolonged amenorrhea from other causes in women who are both human immunodeficiency virus (HIV) seropositive and seronegative. METHODS: This was a cohort study nested in the Women's Interagency HIV Study, a multicenter U.S. study of HIV infection in women. Prolonged amenorrhea was defined as no vaginal bleeding for at least 1 year. A serum follicle stimulating hormone more than 25 milli-International Units/mL and prolonged amenorrhea were used to define ovarian failure. Logistic regressions, chi2, and t tests were performed to estimate relationships between HIV-infection and cofactors with both ovarian failure and amenorrhea from other causes. RESULTS: Results were available for 1,431 women (1,139 HIV seropositive and 292 seronegative). More than one half of the HIV positive women with prolonged amenorrhea of at least 1 year did not have ovarian failure. When adjusted for age, HIV seropositive women were about three times more likely than seronegative women to have prolonged amenorrhea without ovarian failure. Body mass index, serum albumin, and parity were all negatively associated with ovarian failure in HIV seropositive women. CONCLUSION: HIV serostatus is associated with prolonged amenorrhea. It is difficult to ascertain whether the cause of prolonged amenorrhea is ovarian in HIV-infected women without additional testing. LEVEL OF EVIDENCE: II-2. PMID- 17138777 TI - Glucose in vaginal secretions before and after oral glucose tolerance testing in women with and without recurrent vulvovaginal candidiasis. AB - OBJECTIVE: To measure the change of glucose in vaginal secretions during glucose tolerance testing in women with recurrent vulvovaginal candidiasis and in healthy control subjects. METHODS: Thirty-eight women with recurrent vulvovaginal candidiasis and 45 healthy, age-matched controls completed a health questionnaire regarding general and gynecologic health and food and alcohol habits. They all underwent an oral glucose tolerance test and a vaginal examination. Vaginal secretion was collected from the proximal part of the vagina. Glucose in plasma and in vaginal secretions were measured at fasting and after 2 hours and analyzed with the hexokinase method. A sample size analysis showed that the number of subjects included in the study was sufficient for a beta value of 0.80, at the significance level of alpha=.05, at a difference in glucose in vaginal secretions of 30% after oral glucose tolerance test. RESULTS: In healthy women, the median level of glucose in vaginal secretions was 5.2 mM before and 3.7 mM after oral glucose tolerance test, and plasma glucose was 5.0 mM before and 5.8 mM after oral glucose tolerance test. No significant difference was seen regarding change of glucose level in vaginal secretions and plasma glucose after testing, compared with before oral glucose tolerance testing. CONCLUSION: There were no differences between women with recurrent vulvovaginal candidiasis and control subjects regarding change in glucose level in vaginal secretions or in plasma during oral glucose tolerance test. LEVEL OF EVIDENCE: II-2. PMID- 17138778 TI - Regulation of catechol-O-methyltransferase expression in human myometrial cells. AB - OBJECTIVE: The catechol-O-methyltransferase enzyme catalyzes the methylation of the catechol estrogens, 2- or 4-hydroxyestrogen, to 2- or 4-methoxyestrogen. Both the hydroxy estrogens and methoxy estrogens were shown to modulate the effects of estrogen. Because catechol-O-methyltransferase activity controls levels of these metabolites, it may help regulate the cellular estrogenic milieu. In this study, we examined the regulation of catechol-O-methyltransferase expression in human myometrial cells. METHODS: Catechol-O-methyltransferase expression was assessed by reverse transcription-polymerase chain reaction, Western blot, and luciferase assays in human myometrial cells after treatment with estrogen or progesterone. Catechol-O-methyltransferase expression was measured in cells after treatment with tumor necrosis factor alpha (TNFalpha) alone or with lactacystin, a proteasome inhibitor. Luciferase assays were also conducted using human myometrial cells containing an estrogen response element-luciferase reporter gene to measure levels of estrogen-mediated transactivation after treatment with estrogen and increasing concentrations of 2-hydroxestrogen. RESULTS: Catechol-O methyltransferase expression was down-regulated by progesterone or estrogen. Tumor necrosis factor alpha upregulated catechol-O-methyltransferase expression, whereas cotreatment with lactacystin attenuated this response, suggesting that TNFalpha activated nuclear factor kappa B to induce catechol-O-methyltransferase expression. Increased concentrations of 2-hydroxyestrogen attenuated estrogen mediated transcription in the myometrial cells. CONCLUSION: Catechol-O methyltransferase expression may be regulated in the myometrium to control the local action of estrogen. Low levels of catechol-O-methyltransferase in the myometrium would result in an accumulation of 2-hydroxyestrogen and may antagonize the local effect of estrogen. High levels of catechol-O methyltransferase in the myometrium would result in lower levels of 2 hydroxyestrogen and may increase sensitivity to estrogen. PMID- 17138779 TI - Weight gain and spontaneous preterm birth: the role of race or ethnicity and previous preterm birth. AB - OBJECTIVE: To study how the relationship between gestational weight gain and spontaneous preterm birth interacts with maternal race or ethnicity and previous preterm birth status. METHODS: This was a retrospective cohort study of singleton births to women of normal or low prepregnancy body mass index. Gestational weight gain was measured as total weight gain divided by weeks of gestation at delivery, and weight gain was categorized as low (less than 0.27 kg/wk,), normal (0.27-0.52 kg/wk), or high (more than 0.52 kg/wk). Univariable and multivariable analyses were performed on the relationship between weight gain categories and spontaneous preterm birth, stratified by maternal race or ethnicity and history of previous preterm birth. RESULTS: Overall, low weight gain was associated with spontaneous preterm birth (adjusted odds ratio [AOR] 2.5, 95% confidence interval [CI] 2.0 3.1). Although low gain was consistently associated with increased spontaneous preterm birth, some differences were found in subgroup analysis. Among African Americans with a previous preterm birth, both low and high weight gain were associated with increased odds of spontaneous preterm birth (AOR for low weight gain 4.3, 95% CI 1.2-15.5; AOR for high weight gain 6.1, 95% CI 1.8-20.2). For all other groups, high weight gain was not associated with spontaneous preterm birth. Among Asians with a previous preterm birth, low weight gain was not statistically significantly associated with spontaneous preterm birth (AOR 1.9, 95% CI 0.5-7.7). Among Asians there was also a non-statistically significant inverse relationship between high weight gain and spontaneous preterm birth (AOR 0.5, 95% CI 0.3-1.1). CONCLUSION: These results confirm an association between low maternal weight gain and spontaneous preterm birth. The effect modification of maternal race or ethnicity and history of previous preterm birth on this association deserves further study. LEVEL OF EVIDENCE: II-2. PMID- 17138780 TI - Rates of postpartum glucose testing after gestational diabetes mellitus. AB - OBJECTIVE: To estimate rates of postpartum glucose tolerance testing in women diagnosed with gestational diabetes mellitus (GDM) and to assess factors associated with testing. METHODS: This was a retrospective cohort study of 344 women with GDM who received prenatal care in a maternal diabetes clinic during 2001-2004. Rates of postpartum glucose testing were estimated from hospital, clinic, and laboratory records. Demographic, clinical (obstetric history, antenatal, and delivery), and health care information was obtained from chart review. RESULTS: Less than one half (45%) of women with GDM in our cohort underwent postpartum glucose testing-more than one third (36%) of whom had persistent abnormal glucose tolerance. After adjusting for clinical and health care characteristics, there was no independent relationship between most demographic characteristics and postpartum testing. Nor was there an association between clinical characteristics and the likelihood of being tested. Postpartum testing was strongly associated only with attendance of the postpartum visit: 54% of women who attended the visit were tested compared with 17% of women who did not attend (adjusted relative risk 3.04, 95% confidence interval 1.75-5.34, P<.001). CONCLUSION: Although persistent abnormal glucose tolerance was common in our cohort, less than half of the women were tested for it. Our data suggest that to increase rates of postpartum glucose testing, improved attendance at the postpartum visit with greater attention to testing and better continuity between antenatal and postpartum care are required. LEVEL OF EVIDENCE: II-2. PMID- 17138781 TI - Early labor assessment and support at home versus telephone triage: a randomized controlled trial. AB - OBJECTIVE: To compare rates of cesarean delivery among women who were triaged by obstetric nurses, either by telephone or by means of home visits. METHODS: Healthy, nulliparous women in labor at term with uncomplicated pregnancies residing in the City of Vancouver, British Columbia, and suburbs between November 2001 and October 2004 were randomized when they sought advice about when to come to hospital. Women randomized to telephone triage (n=731) were provided with advice by telephone. Women randomized to a home visit (n=728) were triaged after a "hands-on" assessment in their homes. RESULTS: The relative risk (RR) for cesarean delivery among home-triaged women compared with those receiving only telephone support was 1.12 (95% confidence interval [CI] 0.94-1.32). The study was designed to have 80% power to detect a RR less than 0.78 or greater than 1.27 for cesarean delivery. Significantly fewer women in the home visit group were admitted to hospital with cervical dilatation at 3 cm or less (RR 0.85, 95% CI 0.76-0.94). Significantly more women in the home visit group managed their labor without a visit to hospital for assessment (RR 1.54, 95% CI 1.23-1.92). There were no statistically significant differences in use of narcotic analgesia, epidural analgesia, and augmentation of labor. Adverse neonatal outcomes were rare and did not differ between study groups. CONCLUSION: Early labor assessment and support at home versus support by telephone reduces the number of visits to hospital in latent phase labor but does not impact cesarean delivery rates among healthy nulliparous women. CLINICAL TRIAL REGISTRATION: ISRCTN, www.controlled trials.com/isrctn, MCT-44153 LEVEL OF EVIDENCE: I. PMID- 17138782 TI - Validation of a formula that calculates the estimated risk of respiratory distress syndrome. AB - OBJECTIVE: Several groups, including ours, have developed probabilistic models that incorporate both the surfactant-to-albumin ratio (TDx-FLM II) and gestational age to more accurately predict the risk of neonatal respiratory distress syndrome (RDS) and eliminate the current categorical "immature"/"indeterminate"/"mature" interpretation. We validate our model using a separate data set, with the goal of providing the clinician with a risk score. METHODS: The medical records of all women who had TDx-FLM II testing performed at Brigham and Women's Hospital between January 1, 2003, and December 31, 2005, were reviewed to gather a population upon which to validate our previous logistic regression model. Receiver operating characteristic curve and Hosmer-Lemeshow analysis was conducted to determine the performance of our model and another model in this new population. RESULTS: A total of 233 mother-neonate pairs (21 RDS, 212 non-RDS) met criteria for analysis. The receiver operating characteristic analysis illustrated that our previous formula was a strong predictor of the risk of RDS with an area under the curve of 0.902 (95% confidence interval 0.849-0.955). In addition, using the Hosmer-Lemeshow analysis, our formula produced an excellent overall fit (P=.95), whereas another published model was a poor fit to our data (P=.002). CONCLUSION: Our previously derived logistic regression model formula incorporating TDx-FLM II results and gestational age to predict risk of neonatal respiratory distress syndrome was robust and stable over time in an independent data set. The results suggest that the equation can be implemented clinically to assist physicians and patients and used by other institutions after their own internal validation. LEVEL OF EVIDENCE: III. PMID- 17138783 TI - Training for shoulder dystocia: a trial of simulation using low-fidelity and high fidelity mannequins. AB - OBJECTIVE: To evaluate the effectiveness of simulation training for shoulder dystocia management and compare training using a high-fidelity mannequin with that using traditional devices. METHODS: Training was undertaken in six hospitals and a medical simulation center in the United Kingdom. Midwives and obstetricians working for participating hospitals were eligible for inclusion. One hundred forty participants (45 doctors, 95 midwives) were randomized to training with a high-fidelity training mannequin (incorporating force perception training) or traditional low-fidelity mannequins. Performance was assessed pre- and posttraining, using a videoed, standardized shoulder dystocia simulation. Outcome measures were delivery, head-to-body delivery time, use of appropriate and inappropriate actions, force applied, and communication. RESULTS: One hundred thirty-two participants completed the posttraining assessment. All training was associated with improved performance: use of basic maneuvers 114 of 140 (81.4%) to 125 of 132 (94.7%) (P=.002), successful deliveries 60 of 140 (42.9%) to 110 of 132 (83.3%) (P<.001), good communication with the patient 79 of 139 (56.8%) to 109 of 132 (82.6%) (P<.001), pre- and posttraining, respectively. Training with the high-fidelity mannequin was associated with a higher successful delivery rate than training with traditional devices: 94% compared with 72% (odds ratio 6.53, 95% confidence interval 2.05-20.81; P=.002). Total applied force was significantly lower for those who had undergone force training (2,030 Newton seconds versus 2,916 Newton seconds; P=.006) but there was no significant difference in the peak applied force 102 Newtons versus 112 Newtons (P=.242). CONCLUSION: This study verifies the need for shoulder dystocia training; before training only 43% participants could achieve delivery. All training with mannequins improved the management of simulated shoulder dystocia. Training on a high-fidelity mannequin, including force perception teaching, offered additional training benefits. LEVEL OF EVIDENCE: I. PMID- 17138784 TI - Incidence and determinants of peripartum hysterectomy. AB - OBJECTIVE: Most studies of peripartum hysterectomy are conducted in single institutions, limiting the ability to provide national incidence estimates and examine risk factors. The objective of this study was to provide a national estimate of the incidence of peripartum hysterectomy and to examine factors associated with the procedure. METHODS: We used data for 1998-2003 from the Healthcare Cost and Utilization Project Nationwide Inpatient Sample, an annual nationally representative survey of inpatient hospitalizations. Peripartum hysterectomy was defined as a hysterectomy and delivery occurring during the same hospitalization. Odds ratios (ORs) and 95% confidence intervals (CIs) were adjusted for maternal and hospital characteristics using logistic regression. RESULTS: During 1998-2003, an estimated 18,339 peripartum hysterectomies occurred in the United States (0.77 per 1,000 deliveries). Compared with vaginal delivery without a previous cesarean delivery, the ORs of peripartum hysterectomy for other delivery types were as follows: repeat cesarean, 8.90 (95% CI 8.09-9.79); primary cesarean, 6.54 (95% CI 5.95-7.18); and vaginal birth after cesarean, 2.70 (95% CI 2.23-3.26). Multiple births were associated with an increased risk compared with singleton births (OR 1.41, 95% CI 1.16-1.71). CONCLUSION: Our results suggest that vaginal birth after cesarean, primary and repeat cesarean deliveries, and multiple births are independently associated with an increased risk for peripartum hysterectomy. These findings may be of concern, given the increasing rate of both cesarean deliveries and multiple births in the United States. LEVEL OF EVIDENCE: III. PMID- 17138785 TI - Recurrence of clinical chorioamnionitis in subsequent pregnancies. AB - OBJECTIVE: To establish the role of clinical chorioamnionitis as an independent risk factor for recurrence in a subsequent pregnancy. METHOD: This was a historical cohort study of pregnant women who had their first and second deliveries at our institution between January 1988 and May 2005. The index pregnancy was restricted to those who delivered vaginally. Data were collected from a continuously updated obstetric database and included demographic and labor characteristics and neonatal outcomes. Chorioamnionitis was diagnosed clinically. RESULTS: The study population consisted of 23,397 women. During the index pregnancy, 10% of women developed chorioamnionitis. This group was significantly different from the rest of the cohort in terms of age, ethnicity, length of labor, epidural analgesia, use of internal monitors, and incidence of prolonged rupture of membranes. In the second pregnancy, 6% of those women again developed chorioamnionitis compared with 2% of women who did not have chorioamnionitis in the first pregnancy (odds ratio 2.93, 95% confidence interval 2.40-3.57). After adjusting for the above confounders, the increased risk of recurrence persisted (odds ratio 1.85, 95% confidence interval 1.49-2.30). CONCLUSION: Women delivering vaginally who were diagnosed with chorioamnionitis during their first pregnancy are at increased risk for chorioamnionitis in a subsequent pregnancy. This supports the concept that there may be a predisposition to chorioamnionitis that should be further investigated. LEVEL OF EVIDENCE: II-2. PMID- 17138786 TI - Perinatal factors associated with cerebral palsy in children born in Sweden. AB - OBJECTIVE: To identify perinatal factors associated with cerebral palsy (CP). METHODS: This was a case-control study based on the Swedish Medical Birth Registry and the Swedish Hospital Discharge Registry, including 2,303 infants born in Sweden 1984-1998 with a diagnosis of CP and 1.6 million infants without this diagnosis. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated. RESULTS: Infants born preterm had a highly increased risk for CP, and constituted 35% of all cases; OR 34 (95% CI 29-39) in weeks 23-27, OR 37 (95% CI 32-42) in weeks 28-29, OR 26 (95% CI 23-30) in weeks 30-31, and OR 3.9 (95% CI 3.4-4.4) in weeks 32-36. Boys had a higher risk (sex ratio 1.36:1), particularly before term (sex ratio 1.55:1). Other factors associated with CP were being small or large for gestational age at birth, abruptio placentae (OR 8.6, 95% CI 5.6 13.3), maternal insulin-dependent diabetes mellitus type 1 (OR 2.1, 95% CI 1.4 3.1), preeclampsia (OR 1.5, 95% CI 1.3-2.4), being a twin (OR 1.4, 95% CI 1.1 1.6), maternal age older than 40 years (OR 1.4, 95% CI 1.1-1.8) or 35-39 years (OR 1.2, 95% CI 1.1-1.4), primiparity (OR 1.2, 95% CI 1.1-1.3), and smoking (OR 1.2, 95% CI 1.1-1.3). In term infants, low Apgar scores were associated with a high risk for CP; OR 62 (95% CI 52-74) at score 6 at 5 minutes, OR 498 (95% CI 458-542) at score 3. Other factors associated with CP in term infants were breech presentation at vaginal birth (OR 3.0, 95% CI 2.4-3.7), instrumental delivery (OR 1.9, 95% CI 1.6-2.3), and emergency cesarean delivery (OR 1.8, 95% CI 1.6-2.0). CONCLUSION: Preterm birth entails a high risk for CP, but 65% of these children are born at term. Several obstetric factors and low Apgar scores are associated with CP. LEVEL OF EVIDENCE: II-2. PMID- 17138787 TI - Trends in maternal request cesarean delivery from 1991 to 2004. AB - OBJECTIVE: To review definitions and terminology for and to estimate percentages of maternal request cesarean deliveries in the United States between 1991 and 2004. METHODS: National Hospital Discharge Survey data 1991-2004 (N=458,767) were used to identify maternal request cesarean deliveries. After excluding women with a history of cesarean delivery, women who labored, and women with indicated risks against labor, 2,394 potential maternal request cesarean deliveries remained. Indicated risks were identified with a recognized protocol. RESULTS: Maternal request cesarean deliveries have two properties: 1) cesarean delivery before labor and 2) cesarean delivery in the absence of medical conditions presenting a risk for labor. Risk is either absolutely absent or it is relatively absent. In 1991-2004, 0.20% of women who delivered live infants and 1.34% of women who delivered by primary cesarean delivery did so without any medical conditions listed on their hospital discharge record. Estimates for maternal request cesarean deliveries without certain indicated risks were 0.75% for women who delivered live infants and 5.03% for women who delivered by primary cesarean delivery. Maternal request cesarean deliveries without any indicated risk peaked in 1999 for women who delivered live infants. Maternal request cesarean deliveries without certain indicated risks crested in 2004. CONCLUSION: Our estimates were affected by three factors: 1) lack of agreement on a definition of maternal request cesarean deliveries, 2) changes in medical coding practices, and 3) changes in physician response to medical conditions. To validly and reliably estimate maternal request cesarean deliveries requires an empirically tractable, standard definition. LEVEL OF EVIDENCE: III. PMID- 17138788 TI - Cesarean delivery on maternal request: maternal and neonatal outcomes. AB - OBJECTIVE: To review systematically the evidence about maternal and infant outcomes of cesarean delivery on maternal request and planned vaginal delivery. DATA SOURCES: We searched MEDLINE, Cochrane Collaboration resources, and Embase and identified 1,406 articles through dual review using a priori inclusion criteria. METHODS OF STUDY SELECTION: We included English language studies published from 1990 to June 2005 that compared the key reference group (cesarean delivery on maternal request or proxies) and planned vaginal delivery. TABULATION, INTEGRATION, AND RESULTS: We identified 54 articles for maternal and infant outcomes. Virtually no studies exist on cesarean delivery on maternal request, so the knowledge base rests on indirect evidence from proxies with unique and significant limitations. Most studies compared outcomes by actual routes of delivery, resulting in variable relevance to planned routes of delivery. Primary cesarean delivery on maternal request and planned vaginal delivery likely differ with respect to individual outcomes; for instance, risks of urinary incontinence and maternal hemorrhage were lower with planned cesarean, whereas the risk of neonatal respiratory morbidity was higher and maternal length of stay was longer with planned cesarean delivery. However, our comprehensive assessment, across many outcomes, suggests no major differences between primary cesarean delivery on maternal request and planned vaginal delivery, but the evidence is too weak to conclude definitively that differences are completely absent. If a woman chooses to have a cesarean delivery in her first delivery, she is more likely to have subsequent deliveries by cesarean. With increasing numbers of cesarean delivery, risks occur with increasing frequency. CONCLUSION: The evidence is significantly limited by its minimal relevance to primary cesarean delivery on maternal request. Future research requires developing consensus about terminology, creating a minimum data set for cesarean delivery on maternal request, improving study design and statistical analyses, attending to major outcomes and their special measurement issues, assessing both short- and long term outcomes with better measurement strategies, dealing better with confounders, and considering the value or utility of different outcomes. PMID- 17138789 TI - Chronic renal disease in pregnancy. AB - The purpose of this review was to examine the impact of varying degrees of renal insufficiency on pregnancy outcome in women with chronic renal disease. Our search of the literature did not reveal any randomized clinical trials or meta analyses. The available information is derived from opinion, reviews, retrospective series, and limited observational series. It appears that chronic renal disease in pregnancy is uncommon, occurring in 0.03-0.12% of all pregnancies from two U.S. population-based and registry studies. Maternal complications associated with chronic renal disease include preeclampsia, worsening renal function, preterm delivery, anemia, chronic hypertension, and cesarean delivery. The live birth rate in women with chronic renal disease ranges between 64% and 98% depending on the severity of renal insufficiency and presence of hypertension. Significant proteinuria may be an indicator of underlying renal insufficiency. Management of pregnant women with underlying renal disease should ideally entail a multidisciplinary approach at a tertiary center and include a maternal-fetal medicine specialist and a nephrologist. Such women should receive counseling regarding the pregnancy outcomes in association with maternal chronic renal disease and the effect of pregnancy on renal function, especially within the ensuing 5 years postpartum. These women will require frequent visits and monitoring of renal function during pregnancy. Women whose renal disease is further complicated by hypertension should be counseled regarding the increased risk of adverse outcome and need for blood pressure control. Some antihypertensives, especially angiotensin-converting enzyme inhibitors and angiotensin-receptor blockers, should be avoided during pregnancy, if possible, because of the potential for both teratogenic (hypocalvaria) and fetal effects (renal failure, oliguria, and demise). PMID- 17138790 TI - Chronic abdominal pain after laparoscopic sterilization clip placement. PMID- 17138791 TI - Refusals by pharmacists to dispense emergency contraception: a critique. PMID- 17138792 TI - Valacyclovir prophylaxis to prevent recurrent herpes at delivery: a randomized clinical trial. PMID- 17138793 TI - Characteristics of physicians who frequently act as expert witnesses in neurologic birth injury litigation. PMID- 17138794 TI - Performance of contraceptive patch compared with oral contraceptive pill in a high-risk population. PMID- 17138795 TI - A modified technique to deliver the posterior arm in severe shoulder dystocia. PMID- 17138796 TI - Intrapartum electronic fetal heart rate monitoring and the prevention of perinatal brain injury. PMID- 17138797 TI - ACOG educational and practice bulletins: list of titles--December 2006. PMID- 17138798 TI - ACOG committee opinions: list of titles. PMID- 17138799 TI - ACOG Committee Opinion No. 352: Innovative practice: ethical guidelines. AB - Innovations in medical practice are critical to the advancement of medicine. Good clinicians constantly adapt and modify their clinical approaches in ways they believe will benefit patients. Innovative practice frequently is approached very differently from formal research, which is governed by distinct ethical and regulatory frameworks. Although opinions differ on the distinction between research and innovative practice, the production of generalizable knowledge is one defining characteristic of research. Physicians considering innovative practice must disclose to patients the purpose, benefits, and risks of the proposed treatment, including risks not quantified but plausible. They should attempt an innovative procedure only when familiar with and skilled in its basic components. A clinician should share results, positive or negative, with colleagues and, when feasible, teach successful techniques and procedures to other physicians. Practitioners should be wary of adopting innovative procedures or diagnostic tests on the basis of promotions and marketing when the value of the procedures or tests has not been proved. A practitioner should move an innovative practice into formal research if the innovation represents a significant departure from standard practice, if the innovation carries unknown or potentially significant risks, or if the practitioner's goal is to use data from the innovation to produce generalizable knowledge. If there is any question whether innovative practices should be formalized as research, clinicians should seek advice from the relevant institutional review board. PMID- 17138800 TI - ACOG Committee Opinion No. 353: Medical emergency preparedness. AB - Patient care emergencies may occur at any time in a hospital or an outpatient setting. To respond to these emergencies, it is important that obstetrician gynecologists prepare themselves by assessing potential emergencies that might occur, creating plans that include establishing early warning systems, designating specialized first responders, conducting emergency drills, and debriefing staff after actual events to identify what went well and what are opportunities for improvement. Having such systems in place may reduce or prevent the severity of medical emergencies. PMID- 17138801 TI - ACOG Committee Opinion No. 354: Treatment with selective serotonin reuptake inhibitors during pregnancy. AB - Depression is a common condition among women of reproductive age, and selective serotonin reuptake inhibitors (SSRIs) are frequently used for the treatment of depression. However, recent reports regarding SSRI use during pregnancy have raised concerns about fetal cardiac defects, newborn persistent pulmonary hypertension, and other negative effects. The potential risks associated with SSRI use throughout pregnancy must be considered in the context of the risk of relapse of depression if maintenance treatment is discontinued. The American College of Obstetricians and Gynecologists' Committee on Obstetric Practice recommends that treatment with all SSRIs or selective norepinephrine reuptake inhibitors or both during pregnancy be individualized and paroxetine use among pregnant women or women planning to become pregnant be avoided, if possible. PMID- 17138802 TI - ACOG Committee Opinion No. 355: Vaginal agenesis: diagnosis, management, and routine care. AB - Vaginal agenesis occurs in 1 of every 4,000-10,000 females. The most common cause of vaginal agenesis is congenital absence of the uterus and vagina, which also is referred to as mullerian aplasia, mullerian agenesis, or Mayer-Rokitansky-Kuster Hauser syndrome. The condition usually can be successfully managed nonsurgically with the use of successive dilators if it is correctly diagnosed and the patient is sufficiently motivated. Besides correct diagnosis, effective management also includes evaluation for associated congenital renal or other anomalies and careful psychologic preparation of the patient before any treatment or intervention. If surgery is preferred, a number of approaches are available; the most common is the Abbe-McIndoe operation. Women who have a history of mullerian agenesis and have created a functional vagina require routine gynecologic care and can be considered in a similar category to that of women without a cervix and thus annual cytologic screening for cancer may be considered unnecessary in this population. PMID- 17138803 TI - ACOG Committee Opinion No. 356: Routine cancer screening. AB - Obstetrician-gynecologists serve as primary care physicians for many women. Because the obstetrician-gynecologist may be the only physician providing routine care, clinicians should be able to provide recommendations for routine cancer screenings, including those for nongynecologic cancers. This document summarizes recommendations of the American College of Obstetricians and Gynecologists for routine cancer screening for the average-risk American woman. The obstetrician gynecologist should discuss both benefits and limitations of screening tests with the patient. PMID- 17138804 TI - ACOG Committee Opinion No. 357: Primary and preventive care: periodic assessments. AB - Periodic assessments offer an excellent opportunity for obstetricians and gynecologists to provide preventive screening, evaluation, and counseling. This Committee Opinion provides the recommendations of the American College of Obstetricians and Gynecologists' Committee on Gynecologic Practice for routine assessments in primary and preventive care for women based on age and risk factors. PMID- 17138805 TI - From depression to where are my keys: unlocking the behavioral disorders of old age. PMID- 17138806 TI - Population pharmacokinetics in geriatric psychiatry. AB - Although geriatric patients are the major recipients of drugs, most research during drug development is conducted in healthy younger adults. Safe and effective drug therapy in the elderly requires an understanding of both drug disposition and response in older individuals. One of the major issues in studying the elderly relates to the ability to study a large number of people in a minimally invasive way. Population pharmacokinetics can be used to model drug concentrations from a large population of sparsely sampled individuals. Population pharmacokinetics characterizes both the interindividual (between subject) and intraindividual (within-subject) variability, and can identify factors that contribute to pharmacokinetic and pharmacodynamic variability. Population pharmacokinetics can be used to aid in designing large clinical trials by simulating virtual data based on the study design. It can also be used to assess consistency of drug exposure and evaluate its effect on clinical outcome. This article reviews the methods used in pharmacokinetic modeling, as well as providing examples of population pharmacokinetic modeling, highlighting its application to geriatric psychiatry. PMID- 17138807 TI - Investigation of age-related cognitive decline using mice as a model system: behavioral correlates. AB - OBJECTIVE: With recent advances in molecular genetics, mouse models have been generated for a number of disease states. Recently, the authors and others have begun to examine normal age-related cognitive decline using mice as a model system. In this article, and the companion article that follows, the authors present data intended to better parameterize the aging phenotype in mice and examine the possible underlying neuronal mechanisms with special emphasis on age related changes in calcium homeostasis. METHODS: Young (4-6-month-old) and aged (22-24-month-old) C57BL/6 mice were analyzed in terms of their spatial learning abilities in the hidden platform version of the Morris water maze and the delay win-shift version of the Olton radial arm maze. RESULTS: Although aged mice exhibited cognitive impairments in both behavioral tasks used, the extent of impairment differed between the two tasks, which might prove to be advantageous under certain experimental settings. CONCLUSIONS: Like in other areas of biomedical research, mice have become an invaluable research tool in the investigation of learning and memory. It is expected that similar benefits can be realized by developing mouse models for age-related cognitive decline. PMID- 17138808 TI - Investigation of age-related cognitive decline using mice as a model system: neurophysiological correlates. AB - OBJECTIVE: Learning and memory impairments without overt pathology often accompany advancing age. To gain a better understanding of the underlying neuronal substrates associated with this age-related cognitive decline, the authors have begun to use mice as an animal model system. As described in the companion paper, mice exhibit age-related impairments in cognition. Here, the authors explore the possibility that age-related changes in neuronal function may be the result of deregulation of cytosolic free calcium homeostasis. METHODS: Calcium homeostasis in young and aged mice was examined by measuring the slow afterhyperpolarization (sAHP) in hippocampal neurons as well as assessing voltage dependent calcium channel mediated long-term potentiation (vdccLTP). In addition, putative changes in phosphorylation of the L-type channel Ca(V)1.2 by cAMP dependent protein kinase were examined. RESULTS: Both neurophysiological measures of calcium homeostasis indicated an increase in activity-dependent calcium influx. This increase was not the result of an age-related increase in phosphorylation of the L-type channel Ca(V)1.2 by cAMP-dependent protein kinase. CONCLUSIONS: Like in other areas of biomedical research, mice have become an invaluable research tool in the investigation of learning and memory. It is expected that similar benefits can be realized by developing mouse models for age related cognitive decline. PMID- 17138809 TI - Vitamin D deficiency is associated with low mood and worse cognitive performance in older adults. AB - BACKGROUND: Vitamin D deficiency is common in older adults and has been implicated in psychiatric and neurologic disorders. This study examined the relationship among vitamin D status, cognitive performance, mood, and physical performance in older adults. METHODS: A cross-sectional group of 80 participants, 40 with mild Alzheimer disease (AD) and 40 nondemented persons, were selected from a longitudinal study of memory and aging. Cognitive function was assessed using the Short Blessed Test (SBT), Mini-Mental State Exam (MMSE), Clinical Dementia Rating (CDR; a higher Sum of Boxes score indicates greater dementia severity), and a factor score from a neuropsychometric battery; mood was assessed using clinician's diagnosis and the depression symptoms inventory. The Physical Performance Test (PPT) was used to measure functional status. Serum 25 hydroxyvitamin D levels were measured for all participants. RESULTS: The mean vitamin D level in the total sample was 18.58 ng/mL (standard deviation: 7.59); 58% of the participants had abnormally low vitamin D levels defined as less than 20 ng/mL. After adjusting for age, race, gender, and season of vitamin D determination, vitamin D deficiency was associated with presence of an active mood disorder (odds ratio: 11.69, 95% confidence interval: 2.04-66.86; Wald chi(2) = 7.66, df = 2, p = 0.022). Using the same covariates in a linear regression model, vitamin D deficiency was associated with worse performance on the SBT (F = 5.22, df = [2, 77], p = 0.044) and higher CDR Sum of Box scores (F = 3.20, df = [2, 77], p = 0.047) in the vitamin D-deficient group. There was no difference in performance on the MMSE, PPT, or factor scores between the vitamin D groups. CONCLUSIONS: In a cross-section of older adults, vitamin D deficiency was associated with low mood and with impairment on two of four measures of cognitive performance. PMID- 17138810 TI - Plasma C-peptide and cognitive performance in older men without diabetes. AB - BACKGROUND: Emerging evidence suggests that type 2 diabetes may be related to diminished cognition, but little data are available directly regarding the role of insulin levels. OBJECTIVE: The objective of this prospective cohort study was to examine the relation of insulin secretion to cognitive function among men without diabetes. SETTING: The study setting was the Physicians' Health Study U.S. male physicians. PARTICIPANTS: Three hundred sixty-seven men who provided blood samples in 1982, when they had no lifetime history of diabetes and ranged in age from 47-65 years (mean age: 57 years). MEASUREMENTS: The authors assayed plasma C-peptide, reflecting insulin secretion, in the stored blood samples. Beginning in 2001, an average 18 years after blood collection, the authors administered telephone interviews, including tests of general cognition (Telephone Interview of Cognitive Status [TICS]), verbal memory, and category fluency. The authors used regression models to estimate mean differences in cognitive performance across levels of C-peptide controlling for a wide variety of potential confounding factors. RESULTS: On the TICS, men in the top tertile of C-peptide performed significantly worse than those in the bottom (multivariable adjusted mean difference: -1.01 points, 95% confidence interval: -1.78 to -0.24); this apparent impact of C-peptide on cognition was equivalent to the cognitive differences the authors observed between men 6 years apart in age. Performance on the global score (combining results from all the individual tests) and verbal memory score (combining results from four tests of verbal memory) appeared lower among men in the highest C-peptide tertile, but results were not statistically significant. CONCLUSION: Higher midlife insulin secretion may be related to decreased later-life cognitive function, even among men without diabetes. PMID- 17138811 TI - Disability and depression: investigating a complex relation using physical performance measures. AB - OBJECTIVE: The objective of this study was to examine the relation of physical performance measures with depressive symptoms in older men. METHOD: A cross sectional, multivariate comparison of several measures of upper- and lower extremity performance and their relation with depressive symptoms was performed in 2,856 older Japanese American men, aged 71-93 years, who participated in the fourth examination of the Honolulu Heart Program. Depressive symptoms were measured using an 11-item version of Center for Epidemiologic Studies Depression (CES-D) Scale. A score of at least 9 (from a maximum score of 33) is considered clinically significant. Timed functional performance tests, including walking and repeated chair stands, were used to assess lower-extremity performance; handgrip strength was used as an indicator of upper-extremity performance. RESULTS: Two hundred eighty-three participants (9.9%) had a score of 9 or greater on the 11 question CES-D Scale and were considered to be at high risk for depression. Time to walk 10 feet and time to complete five chair stands were significantly longer in those with depressive symptoms, whereas handgrip strength was significantly lower. Only the association of gait speed (time to walk 10 feet) and depressive symptoms remained significant when all physical performance measures were simultaneously included in a multivariate analysis. CONCLUSION: These results demonstrate physical performance measures, particularly gait speed, may be important potential correlates of depression in community-dwelling older men. PMID- 17138812 TI - Incidence and outcome of depressive symptoms in nursing home patients in the Netherlands. AB - OBJECTIVES: To assess incidence and outcome of depressive symptoms among nursing home (NH) patients and to identify clinical characteristics predicting onset and persistence of depressive symptoms. METHODS: Depressive symptoms (GDS>10) and relevant correlates were assessed at baseline and at follow-up (6 months) in 350 NH-patients of 14 Dutch NH's with the Geriatric Depression Scale (GDS). Predictors of onset and persistence were studied using chi-square statistics and multiple logistic regression analyses. RESULTS: The prevalence of depressive symptoms decreased from 41.3% to 28.9% during six months follow-up. The onset of depressive symptoms in those without depressive symptoms at baseline was 4.7%, while the rate of persistence was 63.3%. Persistence of depressive symptoms was more frequent in patients with higher GDS-scores (18-30) at baseline. No baseline characteristics were associated with onset of depressive symptoms. Persistence of depressive symptoms was only associated with more years of education. CONCLUSIONS: Pre-admission factors and transition may largely be responsible for depressive symptoms among nursing home patients. The observed substantial decrease in prevalence of depressive symptoms over six months is largely due to remission of depressive symptoms present at baseline. Adaptation of nursing home patients to pre-admission factors, facilitated by the nursing home environment, may explain this observed decrease of depressive symptoms. Future studies should evaluate interventions targeted at patients with higher GDS-scores (18-30). PMID- 17138816 TI - Activity and safety of bortezomib in multiple myeloma patients with advanced renal failure: a multicenter retrospective study. AB - Patients with multiple myeloma (MM) frequently present with concomitant renal dysfunction, and those requiring dialysis have particularly poor outcomes. Bortezomib is a reversible proteasome inhibitor with significant activity in MM. This retrospective case analysis evaluated the feasibility and activity of bortezomib-based therapy in MM patients (n = 24) requiring dialysis support for advanced renal failure. All but 1 patient were undergoing dialysis at the time of therapy. Patients received bortezomib alone or bortezomib-based combination therapy. Among 20 patients with available response data, overall response rate (complete response [CR] + partial response) was 75%, with 30% CR + near CR. One patient was spared dialysis, and 3 other patients became independent of dialysis following bortezomib-based treatment. These encouraging results suggest that bortezomib or bortezomib-based regimens can be used in MM patients requiring dialysis, with manageable toxicities. Further studies will more formally evaluate the impact of bortezomib-based regimens in this patient population. PMID- 17138817 TI - Dasatinib induces notable hematologic and cytogenetic responses in chronic-phase chronic myeloid leukemia after failure of imatinib therapy. AB - Although imatinib induces marked responses in patients with chronic myeloid leukemia (CML), resistance is increasingly problematic, and treatment options for imatinib-resistant or -intolerant CML are limited. Dasatinib, a novel, highly potent, oral, multitargeted kinase inhibitor of BCR-ABL and SRC family kinases, induced cytogenetic responses in a phase 1 study in imatinib-resistant or intolerant CML and was well tolerated. Initial results are presented from a phase 2 study of 186 patients with imatinib-resistant or -intolerant chronic-phase CML (CML-CP) designed to further establish the efficacy and safety of dasatinib (70 mg twice daily). At 8-months' follow-up, dasatinib induced notable responses, with 90% and 52% of patients achieving complete hematologic and major cytogenetic responses (MCyR), respectively. Responses were long lasting: only 2% of patients achieving MCyR progressed or died. Importantly, comparable responses were achieved by patients carrying BCR-ABL mutations conferring imatinib resistance. Dasatinib also induced molecular responses, reducing BCR-ABL/ABL transcript ratios from 66% at baseline to 2.6% at 9 months. Nonhematologic adverse events were generally mild to moderate, and most cytopenias were effectively managed with dose modifications. Cross-intolerance with imatinib was not evident. To conclude, dasatinib induces notable responses in imatinib-resistant or intolerant CML-CP, is well tolerated, and represents a promising therapeutic option for these patients. This trial was registered at www.clinicaltrials.gov as CA180013. PMID- 17138818 TI - Extended follow-up of methotrexate-free immunosuppression using sirolimus and tacrolimus in related and unrelated donor peripheral blood stem cell transplantation. AB - We assessed the combination of sirolimus and tacrolimus without methotrexate after myeloablative allogeneic stem cell transplantation from 53 matched related donors (MRDs) and 30 unrelated donors (URDs). All patients received cyclophosphamide and total body irradiation conditioning followed by transplantation of mobilized peripheral blood stem cells. The median time to neutrophil engraftment was 14 days. The median time to platelet engraftment was 12 days. No differences between MRD and URD cohorts was noted. The incidence of grade II-IV and III-IV acute graft-versus-host disease (GVHD) were 20.5% and 4.8%. The cumulative incidence of chronic GVHD was 59.1%. There were no differences in acute or chronic GVHD incidence between MRD and URD cohorts. The omission of methotrexate was associated with low transplant-related toxicity, with 30-day and 100-day treatment-related mortality rates of 0% and 4.8%. Relapse free survival at 1 and 2 years was 72.3% and 68.5%, respectively. Overall survival at 1 and 2 years was 77.1% and 72.2%, respectively. There were no differences in relapse-free or overall survival between MRD and URD cohorts. The substitution of sirolimus for methotrexate as GVHD prophylaxis is associated with rapid engraftment, a low incidence of acute GVHD, minimal transplant-related toxicity, and excellent survival. Differences between MRD and URD cohorts are not evident when effective GVHD prophylaxis is used. PMID- 17138819 TI - Sustained thymopoiesis and improvement in functional immunity induced by exogenous KGF administration in murine models of aging. AB - Age-related thymopoietic insufficiency has been proposed to be related to either defects in lymphohematopoietic progenitors or the thymic microenvironment. In this study, we examined whether keratinocyte growth factor (KGF), an epithelial cell-specific growth factor, could increase thymopoietic capacity in aged mice by restoration of the function of thymic epithelial cells (TECs). The thymic cellularity in KGF-treated aged mice increased about 4-fold compared to placebo treated mice, resulting in an equivalent thymic cellularity to young mice. Enhanced thymopoiesis was maintained for about 2 months after a single course of KGF, and sustained improvement was achieved by administration of monthly courses of KGF. With the enhanced thymopoiesis after KGF treatment, the number of naive CD4 T cells in the periphery and T-cell-dependent antibody production improved in aged mice. KGF induced increased numbers of TECs and intrathymic interleukin-7 (IL-7) production and reorganization of cortical and medullary architecture. Furthermore, KGF enhanced thymopoiesis and normalized TEC organization in klotho (kl/kl) mice, a model of premature degeneration and aging, which displays thymopoietic defects. The result suggests that TEC damage is pathophysiologically important in thymic aging, and KGF therapy may be clinically useful in improving thymopoiesis and immune function in the elderly. PMID- 17138820 TI - CD10-MUM1+ follicular lymphoma lacks BCL2 gene translocation and shows characteristic biologic and clinical features. AB - CD10 and MUM1 are representative B cell differentiation markers. Follicular lymphoma (FL) is usually positive for CD10 and negative for MUM1. In this study, however, we compared 22 FLs with peculiar phenotype CD10-MUM1+ with 119 typical CD10+MUM1- FLs. All CD10-MUM1+ FL patients exhibited follicular structure with follicular dendritic meshwork, and a high rate of somatic hypermutation and ongoing mutation, similar to typical FL. However, CD10-MUM1+ FLs were encountered frequently in the elderly compared with CD10+MUM1- typical FLs (67.0 versus 58.7 years, P < .01), showed high grade (grade 3A or 3B) morphology (91% versus 17%, P < .001), diffuse proliferation (59% vs 19%, P < .001), and lacked BCL2/IGH translocation (5% versus 92.5%, P < .001), which is the most characteristic aberration in FL, and 88% showed BCL6 gene abnormalities (translocation or amplification). Our results indicate that CD10-MUM1+ FL is different from typical FL with respect to biologic and clinical features. PMID- 17138821 TI - Results of a randomized international study of high-risk central nervous system B non-Hodgkin lymphoma and B acute lymphoblastic leukemia in children and adolescents. AB - The prognosis for higher risk childhood B-cell non-Hodgkin lymphoma has improved over the past 20 years but the optimal intensity of treatment has yet to be determined. Children 21 years old or younger with newly diagnosed B-cell non Hodgkin lymphoma/B-cell acute lymphoblastic leukemia (B-NHL/B-ALL) with higher risk factors (bone marrow [BM] with or without CNS involvement) were randomized to standard intensity French-American-British/Lymphoma Malignancy B (FAB/LMB) therapy or reduced intensity (reduced cytarabine plus etoposide and deletion of 3 maintenance courses M2, M3, M4). All patients with CNS disease had additional high-dose methotrexate (8 g/m2) plus extra intrathecal therapy. Fifty-one percent had BM involvement, 20% had CNS involvement, and 29% had BM and CNS involvement. One hundred ninety patients were randomized. The probabilities of 4-year event free survival (EFS) and survival (S) were 79% +/- 2.7% and 82% +/- 2.6%, respectively. In patients in remission after 3 cycles who were randomized to standard versus reduced-intensity therapy, the 4-year EFS after randomization was 90% +/- 3.1% versus 80% +/- 4.2% (one-sided P = .064) and S was 93% +/- 2.7% versus 83% +/- 4.0% (one-sided P = .032). Patients with either combined BM/CNS disease at diagnosis or poor response to cyclophosphamide, Oncovin [vincristine], prednisone (COP) reduction therapy had a significantly inferior EFS and S (P < .001). Standard-intensity FAB/LMB therapy is recommended for children with high risk B-NHL (B-ALL with or without CNS involvement). PMID- 17138823 TI - Regression of warfarin-induced medial elastocalcinosis by high intake of vitamin K in rats. AB - Arterial calcification (AC) is generally regarded as an independent risk factor for cardiovascular morbidity and mortality. Matrix Gla protein (MGP) is a potent inhibitor of AC, and its activity depends on vitamin K (VK). In rats, inactivation of MGP by treatment with the vitamin K antagonist warfarin leads to rapid calcification of the arteries. Here, we investigated whether preformed AC can be regressed by a VK-rich diet. Rats received a calcification-inducing diet containing both VK and warfarin (W&K). During a second 6-week period, animals were randomly assigned to receive either W&K (3.0 mg/g and 1.5 mg/g, subsequently), a diet containing a normal (5 microg/g) or high (100 microg/g) amount of VK (either K1 or K2). Increased aortic calcium concentration was observed in the group that continued to receive W&K and also in the group changed to the normal dose of VK and AC progressed. Both the VK-rich diets decreased the arterial calcium content by some 50%. In addition, arterial distensibility was restored by the VK-rich diet. Using MGP antibodies, local VK deficiency was demonstrated at sites of calcification. This is the first study in rats demonstrating that AC and the resulting decreased arterial distensibility are reversible by high-VK intake. PMID- 17138822 TI - IRF-4 and c-Rel expression in antiviral-resistant adult T-cell leukemia/lymphoma. AB - Adult T-cell leukemia/lymphoma (ATLL) is a generally fatal malignancy. Most ATLL patients fare poorly with conventional chemotherapy; however, antiviral therapy with zidovudine (AZT) and interferon alpha (IFN-alpha) has produced long-term clinical remissions. We studied primary ATLL tumors and identified molecular features linked to sensitivity and resistance to antiviral therapy. Enhanced expression of the proto-oncogene c-Rel was noted in 9 of 27 tumors. Resistant tumors exhibited c-Rel (6 of 10; 60%) more often than did sensitive variants (1 of 9; 11%). This finding was independent of the disease form. Elevated expression of the putative c-Rel target, interferon regulatory factor-4 (IRF-4), was observed in 10 (91%) of 11 nonresponders and in all tested patients with c-Rel+ tumors and occurred in the absence of the HTLV-1 oncoprotein Tax. In contrast, tumors in complete responders did not express c-Rel or IRF-4. Gene rearrangement studies demonstrated the persistence of circulating T-cell clones in long-term survivors maintained on antiviral therapy. The expression of nuclear c-Rel and IRF-4 occurs in the absence of Tax in primary ATLL and is associated with antiviral resistance. These molecular features may help guide treatment. AZT and IFN-alpha is a suppressive rather than a curative regimen, and patients in clinical remission should remain on maintenance therapy indefinitely. PMID- 17138824 TI - HGF inhibits BMP-induced osteoblastogenesis: possible implications for the bone disease of multiple myeloma. AB - The bone disease in multiple myeloma is caused by an uncoupling of bone formation from bone resorption. A key difference between patients with and patients without osteolytic lesion is that the latter have fewer and less active osteoblasts. Hepatocyte growth factor (HGF) is often produced by myeloma cells and is found at high concentrations in the bone marrow of patients with multiple myeloma. Here we show that HGF inhibited bone morphogenetic protein (BMP)-induced in vitro osteoblastogenesis. Thus, HGF inhibited BMP-induced expression of alkaline phosphatase in human mesenchymal stem cells (hMSCs) and the murine myoid cell line C2C12, as well as mineralization by hMSCs. Furthermore, the expression of the osteoblast-specific transcription factors Runx2 and Osterix was reduced by HGF treatment. HGF promoted proliferation of hMSCs, and the BMP-induced halt in proliferation was overridden by HGF, keeping the cells in a proliferative, undifferentiating state. BMP-induced nuclear translocation of receptor-activated Smads was inhibited by HGF, providing a possible explanation of how HGF inhibits BMP signaling. The in vitro data were supported by the observation of a negative correlation between HGF and a marker of osteoblast activity, bone-specific alkaline phosphatase (rho = -0.45, P = .008), in sera from 34 patients with myeloma. These observations suggest that HGF inhibits bone formation in multiple myeloma. PMID- 17138825 TI - Hyperacute GVHD: risk factors, outcomes, and clinical implications. AB - Acute graft-versus-host disease (GVHD) is a major limiting factor in allogeneic hematopoietic stem cell transplantation (HSCT), and the timing of acute GVHD may affect patient outcomes. We evaluated the incidence, risk factors, clinical manifestations, and outcomes of hyperacute GVHD, defined as that occurring within 14 days after transplantation, among 809 consecutive HSCTs at the University of Texas M.D. Anderson Cancer Center. Of 265 patients with grade II-IV acute GVHD, 27% had biopsy-proven hyperacute GVHD. Skin involvement was significantly more common (88% versus 44%) and more severe (stage III-IV, 88% versus 66%) in the hyperacute group compared with acute GVHD diagnosed after day 14. On multivariate analysis, significant risk factors for hyperacute GVHD included a mismatched related or matched unrelated donor, a myeloablative conditioning regimen, more than 5 prior chemotherapy regimens, and donor-recipient sex mismatch. Hyperacute GVHD was associated with a significantly lower response rate to first-line therapy and a higher rate of nonrelapse mortality in patients with a mismatched related or matched unrelated donor graft. In conclusion, hyperacute GVHD accounts for a substantial proportion of grade II-IV acute GVHD after HSCT. Patients at high risk or with a diagnosis of hyperacute GVHD should be included in clinical studies. PMID- 17138826 TI - Interferon regulatory factor 3-dependent responses to lipopolysaccharide are selectively blunted in cord blood cells. AB - The synthesis of interferon-beta (IFNbeta) and IFN-inducible factors elicited by lipopolysaccharide (LPS) depends on the transcriptional activity of interferon regulatory factor 3 (IRF-3) downstream of Toll-like receptor-4 (TLR4). To examine the ability of human newborns to mount TLR4-mediated IRF-3-dependent responses, we analyzed the pattern of genes expressed on the addition of LPS to cord blood or cord blood monocyte-derived dendritic cells (moDCs). Expression of IFNbeta and IFN-inducible genes was selectively impaired in neonatal blood and moDCs as compared with their adult counterparts. This selective defect was confirmed by microarray experiments on moDCs. Altered expression of IFN-inducible genes was related to impaired IFNbeta synthesis because IFNbeta signaling was functional in neonatal moDCs. However, addition of exogenous IFNbeta failed to restore LPS induced IL-12p70 synthesis which was previously shown to be defective in neonatal moDCs. Although LPS-induced IRF-3 nuclear translocation was observed both in adult and neonatal moDCs, IRF-3 DNA-binding activity and association with the coactivator CREB-binding protein (CBP) were decreased in neonatal as compared with adult moDCs. We conclude that impaired IRF-3/CBP interaction in neonatal blood cells exposed to LPS is associated with impaired expression of IFNbeta and IFN-inducible genes. Because IRF-3 activity is also required for IL-12p70 synthesis, our findings provide a molecular basis for the decreased ability of LPS-stimulated neonatal moDCs to elicit Th1-type responses. PMID- 17138827 TI - Human herpesvirus 8 acute infection of endothelial cells induces monocyte chemoattractant protein 1-dependent capillary-like structure formation: role of the IKK/NF-kappaB pathway. AB - Human herpesvirus 8 (HHV-8) is considered the causative agent of Kaposi sarcoma, a highly vascularized neoplasm characterized by spindle-shaped cells of endothelial origin and inflammatory cell infiltration. The cell transforming ability of HHV-8 has been associated with the activation of NF-kappaB, a nuclear factor playing a pivotal role in promoting inflammation and cell proliferation; however, little is known about NF-kappaB activation during acute HHV-8 infection. In the present study, we used a recently established in vitro model of HHV-8 acute productive infection in endothelial cells to investigate the effect of HHV 8 on NF-kappaB activity and function. HHV-8 rapidly and potently induced NF kappaB activity in endothelial cells via stimulation of the IkappaB kinase (IKK). Following IKK activation, HHV-8 selectively triggered the production of high levels of monocyte chemoattractant protein 1 (MCP-1), whereas it did not affect the expression of other NF-kappaB-dependent proinflammatory proteins, including TNF-alpha, IL-8, and RANTES. Deletion of NF-kappaB-binding sites in the MCP-1 enhancer resulted in significant inhibition of HHV-8-induced transcription. Furthermore, MCP-1 production was accompanied by virus-induced capillary-like structure formation at early stages of infection. The results suggest that HHV-8 induced MCP-1 may play an important role in promoting inflammation and pathogenic angiogenesis typical of HHV-8-associated lesions. PMID- 17138828 TI - The effect of deoxygenation on whole-cell conductance of red blood cells from healthy individuals and patients with sickle cell disease. AB - Red blood cells from patients with sickle cell disease (SCD) exhibit increased electrogenic cation permeability, particularly following deoxygenation and hemoglobin (Hb) polymerisation. This cation permeability, termed P(sickle), contributes to cellular dehydration and sickling, and its inhibition remains a major goal for SCD treatment. Nevertheless, its characteristics remain poorly defined, its molecular identity is unknown, and effective inhibitors have not been established. Here, patch-clamp methodology was used to record whole-cell currents in single red blood cells from healthy individuals and patients with SCD. Oxygenated normal red blood cells had a low membrane conductance, unaffected by deoxygenation. Oxygenated HbS cells had significantly increased conductance and, on deoxygenation, showed a further rise in membrane conductance. The deoxygenation-induced pathway was variable in magnitude. It had equal permeability to Na(+) and K(+), but was less permeable to NMDG(+) and Cl(-). Conductance to Ca(2+) was also of a similar magnitude to that of monovalent cations. It was inhibited by DIDS (100 microM), Zn(2+) (100 microM), and by Gd(3+) (IC(50) of approximately 2 microM). It therefore shares some properties with P(sickle). These findings represent the first electrical recordings of single HbS cells and will facilitate progress in understanding altered red blood cell cation transport characteristics of SCD. PMID- 17138829 TI - Defective apoptosis of peripheral-blood lymphocytes in hyper-IgD and periodic fever syndrome. AB - Hereditary periodic fever syndromes are characterized by incapacitating attacks of fever and generalized inflammation. While the mutated genes for the major syndromes in this group are known, the pathogenesis remains unclear. The aim of this study was to investigate apoptosis in patients with periodic fever as a possible pathogenic factor. We measured anisomycin-induced apoptosis with annexin V flow cytometry and caspase-3/7 activity in peripheral-blood lymphocytes from symptom-free patients with hyper-IgD and periodic fever syndrome (HIDS; n = 10), TNF-receptor-associated periodic syndrome (TRAPS; n = 7), and familial Mediterranean fever (FMF; n = 2). HIDS lymphocytes showed a decreased percentage of apoptosis during remission by both methods compared with controls (17.8% vs 55.4%), whereas no difference was observed in TRAPS or FMF lymphocytes. This defective apoptosis of lymphocytes may be a central pathogenic mechanism in HIDS, since dysfunction of one of the inhibitory mechanisms to curtail the immunologic response could cause an unbridled generalized inflammation after a trivial stimulus. PMID- 17138830 TI - Going to extremes of lung volume. PMID- 17138831 TI - Tracheal stenosis: a flow dynamics study. AB - Patients referred for treatment of tracheal stenosis typically are asymptomatic until critical narrowing of the airway occurs, which then requires immediate intervention. To understand how tracheal stenosis affects local pressure drops and explore how a dramatic increase in pressure drop could possibly be detected at an early stage, a computational fluid dynamics (CFD) study was undertaken. We assessed flow patterns and pressure drops over tracheal stenoses artificially inserted into a realistic three-dimensional upper airway model derived from multislice computed tomography images obtained in healthy men. Solving the Navier Stokes equations (with a Yang-shih k-epsilon turbulence model) for different degrees of tracheal constriction located approximately one tracheal diameter below the glottis, the simulated pressure drop over the stenosis (DeltaP) was seen to dramatically increase only when well over 70% of the tracheal lumen was obliterated. At 30 l/min, DeltaP increased from 7 Pa for a 50% stenosis to, respectively, 46 and 235 Pa for 80% and 90% stenosis. The pressure-flow relationship in the entire upper airway model (between mouth and end of trachea) in the flow range 0-60 l/min showed a power law relationship with best-fit flow exponent of 1.77 in the absence of stenosis. The exponent became 1.92 and 2.00 in the case of 60% and 85% constriction, respectively. The present simulations confirm that the overall pressure drop at rest is only affected in case of severe constriction, and the simulated flow dependence of pressure drop suggests a means of detecting stenosis at a precritical stage. PMID- 17138832 TI - Carbohydrate ingestion augments L-carnitine retention in humans. AB - Maintaining hyperinsulinemia (approximately 150 mU/l) during steady-state hypercarnitinemia (approximately 550 micromol/l) increases skeletal muscle total carnitine (TC) content by approximately 15% within 5 h. The present study aimed to investigate whether an increase in whole body carnitine retention can be achieved through L-carnitine feeding in conjunction with a dietary-induced elevation in circulating insulin. On two randomized visits (study A), eight men ingested 3 g/day L-carnitine followed by 4 x 500-ml solutions, each containing flavored water (Con) or 94 g simple sugars (glucose syrup; CHO). In addition, 14 men ingested 3 g/day L-carnitine followed by 2 x 500 ml of either Con or CHO for 2 wk (study B). Carbohydrate ingestion in study A resulted in a fourfold greater serum insulin area under the curve when compared with Con (P < 0.001) and in a lower plasma TC concentration throughout the CHO visit (P < 0.05). Twenty-four hour urinary TC excretion in the CHO visit was lower than in the Con visit in study A (155.0 +/- 10.7 vs. 212.1 +/- 17.2 mg; P < 0.05). In study B, daily urinary TC excretion increased after 3 days (65.9 +/- 18.0 to 281.0 +/- 35.0 mg; P < 0.001) and remained elevated throughout the Con trial. During the CHO trial, daily urinary TC excretion increased from a similar basal value of 53.8 +/- 9.2 to 166.8 +/- 17.3 mg after 3 days (P < 0.01), which was less than during the Con trial (P < 0.01), and it remained lower over the course of the study (P < 0.001). The difference in plasma TC concentration in study A and 24-h urinary TC excretion in both studies suggests that insulin augmented the retention of carnitine in the CHO trials. PMID- 17138833 TI - Effects of heterogeneities on the partitioning of airway and tissue properties in normal mice. AB - We measured the mechanical properties of the respiratory system of C57BL/6 mice using the optimal ventilation waveform method in closed- and open-chest conditions at different positive end-expiratory pressures. The tissue damping (G), tissue elastance (H), airway resistance (Raw), and hysteresivity were obtained by fitting the impedance data to three different models: a constant phase model by Hantos et al. (Hantos Z, Daroczy B, Suki B, Nagy S, Fredberg JJ. J Appl Physiol 72: 168-178, 1992), a heterogeneous Raw model by Suki et al. (Suki B, Yuan H, Zhang Q, Lutchen KR. J Appl Physiol 82: 1349-1359, 1997), and a heterogeneous H model by Ito et al. (Ito S, Ingenito EP, Arold SP, Parameswaran H, Tgavalekos NT, Lutchen KR, Suki B. J Appl Physiol 97: 204-212, 2004). Both in the closed- and open-chest conditions, G and hysteresivity were the lowest and Raw the highest in the heterogeneous Raw model, and G and H were the largest in the heterogeneous H model. Values of G, Raw, and hysteresivity were significantly higher in the closed-chest than in the open-chest condition. However, H was not affected by the conditions. When the tidal volume of the optimal ventilation waveform was decreased from 8 to 4 ml/kg in the closed-chest condition, G and hysteresivity significantly increased, but there were smaller changes in H or Raw. In summary, values of the obtained mechanical properties varied among these models, primarily due to heterogeneity. Moreover, the mechanical parameters were significantly affected by the chest wall and tidal volume in mice. Contribution of the chest wall and heterogeneity to the mechanical properties should be carefully considered in physiological studies in which partitioning of airway and tissue properties are attempted. PMID- 17138834 TI - Chronic upper airway resistive loading induces growth retardation via the GH/IGF I axis in prepubescent rats. AB - The effect of upper airway loading on longitudinal bone growth and various components of the growth hormone (GH)/insulin-like growth factor I (IGF-I) axis has not been fully elucidated. In the present study, the effect of chronic resistive airway loading (CAL) in a prepubescent rat model on linear bone growth and weight gain was investigated. We hypothesize that CAL induced in prepubescent rats will lead to impaired longitudinal growth due to impairment in circulating and liver GH/IGF-I parameters. The tracheae of 22-day-old rats were obstructed by tracheal banding to increase inspiratory esophageal pressure. The GH/IGF-I markers were analyzed using ELISA, RT-PCR, and Western immunoblot analysis 14 days after surgery. Animals exhibited impaired longitudinal growth as demonstrated by reduction of tibia and tail length gains by 40% (P < 0.0001) and body weight gain by 24% (P < 0.0001). No differences were seen in total body energy balance, i.e., oxygen consumption, daily food intake, or arterial blood gases. Circulating GH, IGF-I, and IGF binding protein-3 (IGFBP-3) levels were reduced by 40% (P = 0.037), 30% (P < 0.006), and 27% (P = 0.02), respectively, in the CAL group. Liver IGF-I mRNA level decreased by 20% (P < 0.0002), whereas GH receptor mRNA and protein expression were unchanged. We conclude that impaired longitudinal growth in prepubescent CAL rats is related to a decrease in GH, IGF I, and IGFBP-3 levels. PMID- 17138835 TI - Cardiorespiratory failure in rat induced by severe inspiratory resistive loading. AB - The mechanisms underlying acute respiratory failure induced by respiratory loads are unclear. We hypothesized that, in contrast to a moderate inspiratory resistive load, a severe one would elicit central respiratory failure (decreased respiratory drive) before diaphragmatic injury and fatigue. We also wished to elucidate the factors that predict endurance time and peak tracheal pressure generation. Anesthetized rats breathed air against a severe load ( approximately 75% of the peak tracheal pressure generated during a 30-s occlusion) until pump failure (fall in tracheal pressure to half; mean 38 min). Hypercapnia and hypoxemia developed rapidly ( approximately 4 min), coincident with diaphragmatic fatigue (decreased ratio of transdiaphragmatic pressure to peak integrated phrenic activity) and the detection in blood of the fast isoform of skeletal troponin I (muscle injury). At approximately 23 min, respiratory frequency and then blood pressure fell, followed immediately by secondary diaphragmatic fatigue. Blood taken after termination of loading contained cardiac troponin T (myocardial injury). Contrary to our hypothesis, diaphragmatic fatigue and injury occurred early in loading before central failure, evident only as a change in the timing but not the drive component of the central respiratory pattern generator. Stepwise multiple regression analysis selected changes in mean arterial pressure and arterial Pco(2) during loading as the principal contributing factors in load endurance time, and changes in mean arterial pressure as the principal contributing factor in peak tracheal pressure generation. In conclusion, the temporal development of respiratory failure is not stereotyped but depends on load magnitude; moreover respiratory loads induce cardiorespiratory, not just respiratory, failure. PMID- 17138836 TI - Microinjection of DLH into the region of the caudal ventral respiratory column in the cat: evidence for an endogenous cough-suppressant mechanism. AB - The caudal ventral respiratory column (cVRC) contains premotor expiratory neurons that play an important role in cough-related expiratory activity of chest wall and abdominal muscles. Microinjection of d,l-homocysteic acid (DLH) was used to test the hypothesis that local activation of cVRC neurons can suppress the cough reflex. DLH (20-50 mM, 10-30 nl) was injected into the region of cVRC in nine anesthetized spontaneously breathing cats. Repetitive coughing was elicited by mechanical stimulation of the intrathoracic airways. Electromyograms (EMG) were recorded bilaterally from inspiratory parasternal and expiratory transversus abdominis (ABD) and unilaterally from laryngeal posterior cricoarytenoid and thyroarytenoid muscles. Unilateral microinjection of DLH (1-1.5 nmol) elicited bilateral increases in tonic and phasic respiratory ABD EMG activity, and it altered the respiratory pattern and laryngeal motor activities. However, DLH also decreased cough frequency by 51 +/- 7% compared with control (P < 0.001) and the amplitude of the contralateral (-35 +/- 3%; P < 0.001) and ipsilateral (-34 +/- 5%; P < 0.001) ABD EMGs during postinjection coughs compared with control. The cough alterations were much less pronounced after microinjection of a lower dose of DLH (0.34-0.8 nmol). No cough depression was observed after microinjections of vehicle. These results suggest that an endogenous cough suppressant neuronal network in the region of the cVRC may exist, and this network may be involved in the control of cough reflex excitability. PMID- 17138837 TI - Developmental signals do not further accentuate nonuniform postpneumonectomy compensatory lung growth. AB - Mechanical forces imposed on lung tissue constitute major stimuli for normal lung development and postpneumonectomy (PNX) compensatory growth and remodeling. Superimposing developmental signals on PNX signals augments compensatory alveolar growth but exaggerates airway-parenchymal dissociation (i.e., dysanaptic lung growth); the latter tends to offset benefits derived from the former. In adult dogs after PNX, lobar expansion and growth of the remaining lobes were markedly non-uniform (Ravikumar et al. J Appl Physiol 97:1567-1574, 2004). We hypothesized that superimposing developmental and post-PNX signals further accentuates nonuniformity of lobar growth. We used high-resolution computed tomography (HRCT) to follow regional lung expansion and growth in foxhounds undergoing right PNX at 2.5 mo of age compared with litter-matched control (Sham) animals; scans were performed 4 and 10 mo following surgery, i.e., before and after somatic maturity. Air and tissue volumes were measured in each lobe; tissue volume estimated by HRCT includes air-free tissue and blood in small vessels <1 mm. Interlobar nonuniformity of tissue volume was absent at 4 mo but evident 10 mo after PNX; growth of the remaining left lower lobe gradually lagged behind other lobes. At maturity, nonuniformity of lobar growth in pneumonectomized puppies was similar to that previously reported in pneumonectomized adults. We conclude that superimposing developmental and post-PNX signals enhances some aspects of compensatory lung growth and remodeling without altering its nonuniform spatial distribution. PMID- 17138838 TI - Left ventricular function during arm exercise: influence of leg cycling and lower body positive pressure. AB - The purpose of this study was to characterize left ventricular (LV) diastolic filling and systolic performance during graded arm exercise and to examine the effects of lower body positive pressure (LBPP) or concomitant leg exercise as means to enhance LV preload in aerobically trained individuals. Subjects were eight men with a mean age (+/-SE) of 26.8 +/- 1.2 yr. Peak exercise testing was first performed for both legs [maximal oxygen uptake (Vo(2)) = 4.21 +/- 0.19 l/min] and arms (2.56 +/- 0.16 l/min). On a separate occasion, LV filling and ejection parameters were acquired using non-imaging scintography using in vivo red blood cell labeling with technetium 99(m) first during leg exercise performed in succession for 2 min at increasing grades to peak effort. Graded arm exercise (at 30, 60, 80, and 100% peak Vo(2)) was performed during three randomly assigned conditions: control (no intervention), with concurrent leg cycling (at a constant 15% leg maximal Vo(2)) or with 60 mmHg of LBPP using an Anti G suit. Peak leg exercise LV ejection fraction was higher than arm exercise (60.9 +/- 1.7% vs. 55.9 +/- 2.7%; P < 0.05) as was peak LV end-diastolic volume was reported as % of resting value (110.3 +/- 4.4% vs. 97 +/- 3.7%; P < 0.05) and peak filling rate (end-diastolic volume/s; 6.4 +/- 0.28% vs. 5.2 +/- 0.25%). Concomitant use of either low-intensity leg exercise or LBPP during arm exercise failed to significantly increase LV filling or ejection parameters. These observations suggest that perturbations in preload fail to overcome the inherent hemodynamic conditions present during arm exercise that attenuate LV performance. PMID- 17138839 TI - Intensity of eccentric exercise, shift of optimum angle, and the magnitude of repeated-bout effect. AB - This study compared the effect of four different intensities of initial eccentric exercise (ECC1) on optimum angle shift and extent of muscle damage induced by subsequent maximal eccentric exercise. Fifty-two male students were placed into 100%, 80%, 60%, or 40% groups (n = 13 per group), performing 30 eccentric actions of the elbow flexors of 100%, 80%, 60%, or 40% of maximal isometric strength [maximal voluntary contraction (MVC)] for ECC1, followed 2-3 wk later by a similar exercise (ECC2) that used 100% MVC load. MVC at six elbow joint angles, range of motion, upper arm circumference, serum creatine kinase activity, myoglobin concentration, and muscle soreness were measured before and for 5 days following ECC1 and ECC2. A rightward shift of optimum angle following ECC1 was significantly (P < 0.05) greater for the 100% and 80% than for the 60% and 40% groups, and it decreased significantly (P < 0.05) from immediately to 5 days postexercise. By the time ECC2 was performed, only the 100% group kept a significant shift (4 degrees). Changes in most of the criterion measures following ECC1 were significantly greater for the 100% and 80% groups compared with the 60% and 40% groups. Changes in the criterion measures following ECC2 were significantly (P < 0.05) greater for the 40% group compared with other groups. Although the magnitude of repeated bout effect following ECC2 was significantly (P < 0.05) smaller for the 40% and 60% groups, all groups showed significantly (P < 0.05) reduced changes in criterion measures following ECC2 compared with the ECC1 100% bout. We conclude that the repeated-bout effect was not dependent on the shift of optimum angle. PMID- 17138840 TI - Carbon dioxide and pH effects on temperature-sensitive and -insensitive hypothalamic neurons. AB - The preoptic-anterior hypothalamus (POAH) controls body temperature, and thermoregulatory responses are impaired during hypercapnia. If increased CO(2) or its accompanying acidosis inhibits warm-sensitive POAH neurons, this could provide an explanation for thermoregulatory impairment during hypercapnia. To test this possibility, extracellular electrophysiological recordings determined the effects of CO(2) and pH on the firing rates of both temperature-sensitive and -insensitive neurons in hypothalamic tissue slices from 89 male Sprague-Dawley rats. Firing rate activity was recorded in 121 hypothalamic neurons before, during, and after changing the CO(2) concentration aerating the tissue slice chamber or changing the pH of the solution bathing the tissue slices. Increasing the aeration CO(2) concentration from 5% (control) to 10% (hypercapnic) had no effect on most (i.e., 69%) POAH temperature-insensitive neurons; however, this hypercapnia inhibited the majority (i.e., 59%) of warm-sensitive neurons. CO(2) affected similar proportions of (non-POAH) neurons in other hypothalamic regions. These CO(2) effects appear to be due to changes in pH since the CO(2)-affected neurons responded similarly to isocapnic acidosis (i.e., normal CO(2) and decreased pH) but were not responsive to isohydric hypercapnia (i.e., increased CO(2) and normal pH). These findings may offer a neural explanation for some heat related illnesses (e.g., exertional heat stroke) where impaired heat loss is associated with acidosis. PMID- 17138841 TI - Pioglitazone inhibits androgen production in NCI-H295R cells by regulating gene expression of CYP17 and HSD3B2. AB - Thiazolidinediones (TZDs) such as pioglitazone and rosiglitazone are widely used as insulin sensitizers in the treatment of type 2 diabetes. In diabetic women with polycystic ovary syndrome, treatment with pioglitazone or rosiglitazone improves insulin resistance and hyperandrogenism, but the mechanism by which TZDs down-regulate androgen production is unknown. Androgens are synthesized in the human gonads as well as the adrenals. We studied the regulation of androgen production by analyzing the effect of pioglitazone and rosiglitazone on steroidogenesis in human adrenal NCI-H295R cells, an established in vitro model of steroidogenesis of the human adrenal cortex. Both TZDs changed the steroid profile of the NCI-H295R cells and inhibited the activities of P450c17 and 3betaHSDII, key enzymes of androgen biosynthesis. Pioglitazone but not rosiglitazone inhibited the expression of the CYP17 and HSD3B2 genes. Likewise, pioglitazone repressed basal and 8-bromo-cAMP-stimulated activities of CYP17 and HSD3B2 promoter reporters in NCI-H295R cells. However, pioglitazone did not change the activity of a cAMP-responsive luciferase reporter, indicating that it does not influence cAMP/protein kinase A/cAMP response element-binding protein pathway signaling. Although peroxisome proliferator-activated receptor gamma (PPARgamma) is the nuclear receptor for TZDs, suppression of PPARgamma by small interfering RNA technique did not alter the inhibitory effect of pioglitazone on CYP17 and HSD3B2 expression, suggesting that the action of pioglitazone is independent of PPARgamma. On the other hand, treatment of NCI-H295R cells with mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) inhibitor 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one (PD98059) enhanced promoter activity and expression of CYP17. This effect was reversed by pioglitazone treatment, indicating that the MEK/ERK signaling pathway plays a role in regulating androgen biosynthesis by pioglitazone. PMID- 17138842 TI - Clinical trial of exercise for shoulder pain in chronic spinal injury. AB - BACKGROUND AND PURPOSE: The high prevalence of shoulder pain in wheelchair users may be related to the repetitive use of the upper limbs during self-care and wheelchair-related activities. The purpose of this study was to determine the effects of a controlled 8-week, scapula-focused exercise intervention on pain and functional disability in people with spinal cord injury (SCI) and shoulder impingement symptoms. SUBJECTS: Forty-one manual wheelchair users (with SCI and spina bifida), both with (n=21) and without (n=20) shoulder impingement symptoms, participated. METHODS: The study design was a clinical trial with an asymptomatic control group. Subjects completed the Wheelchair User's Shoulder Pain Index (WUSPI) and the Shoulder Rating Questionnaire (SRQ) and provided patient satisfaction scores at initial and 8-week visits. Subjects in the intervention group were instructed in a home exercise program consisting of stretching and strengthening exercises. Subjects in the asymptomatic control group received no intervention. An analysis of variance model was used to test for group and time effects for the WUSPI, SRQ, and satisfaction scores. RESULTS: Subjects in the intervention group showed significant improvements in all measures as a result of the intervention, whereas asymptomatic control group subjects remained stable. DISCUSSION AND CONCLUSION: A selective 8-week home exercise program is effective in reducing pain and improving function and satisfaction in this population of wheelchair users. PMID- 17138843 TI - Decision-making ability of physical therapists: physical therapy intervention or medical referral. AB - BACKGROUND AND PURPOSE: Opponents of direct access to physical therapy argue that physical therapists may overlook serious medical conditions. More information is needed to determine the ability of physical therapists to practice safely in direct-access environments. The purpose of this study was to describe the ability of physical therapists to make decisions about the management of patients in a direct-access environment. SUBJECTS: Of a random sample of 1,000 members of the Private Practice Section of the American Physical Therapy Association, 394 participated. METHODS: A survey included 12 hypothetical case scenarios. For each case, participants determined whether they would provide intervention without referral, provide intervention and refer, or refer before intervention. The percentage of correct decisions for each group of scenarios was calculated for each participant, and participants were classified as having made correct decisions for 100% of cases or less for each group. Three sets of logistic regressions were completed to determine the characteristics of the participants in relation to the decision category. RESULTS: The average percentages of correct decisions were 87%, 88%, and 79% for musculoskeletal, noncritical medical, and critical medical conditions, respectively. Of all participants, approximately 50% made correct decisions for all cases within each group. The odds of making 100% correct decisions if a physical therapist had an orthopedic specialization were 2.23 (95% confidence interval=1.35-3.71) for musculoskeletal conditions and 1.89 (95% confidence interval=1.14-3.15) for critical medical conditions. DISCUSSION AND CONCLUSION: Physical therapists with an orthopedic specialization were almost twice as likely to make correct decisions for critical medical and musculoskeletal conditions. PMID- 17138844 TI - Gait symmetry and walking speed analysis following lower-extremity trauma. AB - BACKGROUND AND PURPOSE: Gait has been shown to be a major determining factor of function following limb-salvage surgery. However, little is known regarding the measures associated with gait recovery for this patient population. The purpose of this study was to identify clinical measures associated with impaired walking speed and gait asymmetry in patients with lower-extremity reconstruction. SUBJECTS: Study subjects were 381 patients from the Lower Extremity Assessment Project (LEAP) who had undergone reconstruction following severe lower-extremity trauma. METHODS: The LEAP study was a longitudinal study of outcomes following lower-extremity reconstruction. The present study used 24-month clinical follow up data. A combined outcome measure of reduced walking speed and gait deviation was chosen to provide a comprehensive measure of impaired physical mobility. RESULTS: The most significant clinical factors associated with decreased walking speed and gait deviation were impaired ankle plantar-flexion range of motion, knee flexion strength, and a nonreciprocal stair-climbing pattern. DISCUSSION AND CONCLUSION: The findings provide clinicians with specific clinical measures associated with functional recovery in patients with lower-limb reconstruction. These measures, in turn, can be considered to inform treatment decision making and to prioritize interventions. PMID- 17138845 TI - Functional restoration for a chronic lumbar disk extrusion with associated radiculopathy. AB - BACKGROUND AND PURPOSE: The effectiveness of functional restoration (FR) for patients with lumbar disk herniation with associated radiculopathy (LDHR) is unclear. This case report describes how an FR program was used to rehabilitate a patient with such an injury. CASE DESCRIPTION: The patient was a 26-year-old female child care worker with a 12-month history of back pain and a 4-month history of unremitting left leg symptoms. She had clinical and radiological evidence of an L5-S1 disk extrusion with associated left S1 radiculopathy. Interventions. The patient completed a 9-week FR program supervised by a physical therapist. Exercises then were continued more independently for a 2-year period at a public gymnasium. OUTCOMES: Following 9 weeks of supervised FR, the patient demonstrated marked improvement in symptoms and functional ability, and resolution of neurological signs. Fourteen months after commencing FR, a follow up magnetic resonance imaging scan demonstrated resolution of the L5-S1 disk extrusion and relief of S1 nerve root compression. Functional improvements continued and were maintained 2 years following the start of intervention. DISCUSSION: A patient with chronic LDHR who underwent FR made significant improvements. Research is needed to determine the efficacy of an FR approach for treating such patients. PMID- 17138846 TI - An integrated framework for decision making in neurologic physical therapist practice. AB - Decision-making frameworks are used by clinicians to guide patient management, communicate with other health care providers, and educate patients and their families. A number of frameworks have been applied to guide clinical practice, but none are comprehensive in terms of patient management. This article proposes a unifying framework for application to decision making in the management of individuals who have neurologic dysfunction. The framework integrates both enablement and disablement perspectives. The framework has the following attributes: (1) it is patient-centered, (2) it is anchored by the patient/client management model from the Guide for Physical Therapist Practice, (3) it incorporates the Hypothesis-Oriented Algorithm for Clinicians (HOAC) at every step, and (4) it proposes a systematic approach to task analysis for interpretation of movement dysfunction. This framework provides a mechanism for making clinical decisions, developing clinical hypotheses, and formulating a plan of care. Application of the framework is illustrated with a case example of an individual with neurologic dysfunction. PMID- 17138847 TI - Does cardiac rehabilitation improve quality of life for a man with coronary artery disease who received percutaneous transluminal coronary angioplasty with insertion of a stent? PMID- 17138848 TI - Serum prealbumin: Is it a marker of nutritional status or of risk of malnutrition? PMID- 17138849 TI - Efficient and accurate approaches to the laboratory diagnosis of alpha1 antitrypsin deficiency: The promise of early diagnosis and intervention. PMID- 17138850 TI - Will epigenetic allelic ratio analysis turn prenatal diagnosis of trisomy 18 on its EAR? PMID- 17138851 TI - RNAprotect saliva: An optimal room- temperature stabilization reagent for the salivary transcriptome. PMID- 17138852 TI - Measurement of 25-hydroxyvitamin D revisited. PMID- 17138853 TI - Size-exclusion chromatography of circulating cardiac troponin T. PMID- 17138854 TI - A case of pseudoparaproteinemia on capillary zone electrophoresis caused by geloplasma. PMID- 17138855 TI - Maldi-tof mass spectrometry compared with real-time PCR for detection of fetal cell-free DNA in maternal plasma. PMID- 17138856 TI - Pseudoparaproteinemia related to iomeprol administration after angiocardiography: Detection in the beta fraction by capillary zone electrophoresis. PMID- 17138857 TI - Further evidence that the UGT1A1*28 allele is not associated with coronary heart disease: The ECTIM Study. PMID- 17138858 TI - Oxidation of methionine 63 and 83 regulates the effect of S100A9 on the migration of neutrophils in vitro. AB - The calcium-binding proteins S100A8 and S100A9 and their heterocomplex calprotectin are abundant cytosolic constituents in human neutrophils, constitutively expressed by mucosal epithelium and in association with inflammation by epidermal keratinocytes. S100A8 and S100A9 are pleiotropic proteins, which partake in the regulation of leukocyte migration. This study was designed to investigate the effect of S100A9 on neutrophil migration and to explore the mechanisms that regulate this effect. Based on previous results with S100A8, we hypothesized that S100A9 repels neutrophils and that oxidation of S100A9 regulates this function. Using standard Transwell chemotaxis assays and site-directed mutagenesis, we show that S100A9 exerts a chemo-repulsive (fugetactic) effect on peripheral neutrophils, an effect abolished by oxidation of S100A9. After substitution of methionine 63 and 83 for alanine, S100A9 maintained its fugetaxis activity, even in inhibitory, oxidative conditions. Together, the data suggest that S100A9 serves as a molecular switch for oxidative control of inflammation regulated by the oxidation of species-conserved methionine residues. In healthy mucosal tissue, expression of S100A9 by the epithelium may serve to inhibit leukocyte recruitment. However, conditions of oxidative stress, including infection and overgrowth of opportunistic pathogens, may abrogate this activity by neutralizing S100A9 as a result of its oxidative alteration. PMID- 17138859 TI - Interview with Dr. Francisco Sanchez-Madrid regarding Pivotal Advance: CD69 targeting differentially affects the course of collagen-induced arthritis. Interview by Helene F. Rosenberg. PMID- 17138860 TI - Regulation of TNF mediated antiapoptotic signaling in human neutrophils: role of delta-PKC and ERK1/2. AB - TNF is implicated in the suppression of neutrophil apoptosis during sepsis. Multiple signaling pathways are involved in TNF-mediated antiapoptotic signaling; a role for the MAP kinases (MAPK), ERK1/2, and p38 MAPK has been suggested. Antiapoptotic signaling is mediated principally through TNF receptor-1 (TNFR-1), and the PKC isotype-delta (delta-PKC) is a critical regulator of TNFR-1 signaling. delta-PKC associates with TNFR-1 in response to TNF and is required for NFkappaB activation and inhibition of caspase 3. The role of delta-PKC in TNF mediated activation of MAPK is not known. The purpose of this study was to determine whether the MAPK, ERK1/2, and p38 MAPK are involved in TNF antiapoptotic signaling and whether delta-PKC is a key regulator of MAPK activation by TNF. In human neutrophils, TNF activated both p38 MAPK and ERK1/2 principally via TNFR-1. The MEK1/2 inhibitors PD098059 and U0126, but not the p38 MAPK inhibitor SB203580, decreased TNF antiapoptotic signaling as measured by caspase 3 activity. A specific delta-PKC antagonist, V1.1delta-PKC-Tat peptide, inhibited TNF-mediated ERK1/2 activation, but not p38 MAPK. ERK1/2 inhibition did not alter recruitment of delta-PKC to TNFR-1, indicating delta-PKC is acting upstream of ERK1/2. In HL-60 cells differentiated to a neutrophilic phenotype, delta-PKC depletion by delta-PKC siRNA resulted in inhibition of TNF mediated ERK1/2 activation but not p38 MAPK. Thus, ERK1/2, but not p38 MAPK, is an essential component of TNF-mediated antiapoptotic signaling. In human neutrophils, delta-PKC is a positive regulator of ERK1/2 activation via TNFR-1 but has no role in p38 MAPK activation. PMID- 17138861 TI - Hepatoprotective activity of liposomal flavonoid against arsenite-induced liver fibrosis. AB - Arsenic, the environmental metalloid toxicant, is known to induce oxidative damage to liver and produce hepatic fibrosis. The theme of our study was to optimize and evaluate the therapeutic efficacy of galactosylated liposomal flavonoidal antioxidant, quercetin (QC), in combating arsenic-induced hepatic fibrogenesis. The rats of the hepatic damage group were injected s.c. a single dose of sodium arsenite (NaAsO(2)) (100.06 microM/kg b. wt. in 0.5 ml of physiological saline). Hepatocytes and stellate cells were separated. Mitochondrial membranes were isolated from all those separated cells. Oxidative damage was monitored at different isolated subcellular parts of different hepatic cells. Liver fibrosis was also induced by the injection of NaAsO(2). Galactosylated liposomal QC injection before NaAsO(2) treatment checked fibrogenesis completely by protecting the liver from oxidative attack in cellular and subcellular levels. The maximal protections from hepatocellular and fatty metamorphosis, necrosis, Kupffer cell hyperplasia, fibrosis, and in the deposition of collagen contents were observed and reconfirmed by our histopathological and histochemical analysis when rats were treated with galactosylated liposomal QC before NaAsO(2) injection. Application of galactosylated liposomal QC may be a potent therapeutic approach for NaAsO(2) induced fibrogenesis through a complete protection against oxidative attack in cellular and subcellular parts of rat liver. PMID- 17138862 TI - Pharmacokinetics and pharmacodynamics of gamma-hydroxybutyric acid during tolerance in rats: effects on extracellular dopamine. AB - gamma-Hydroxybutyrate (GHB) is a potent sedative/hypnotic and drug of abuse. Tolerance develops to GHB's sedative/hypnotic effects. It is hypothesized that GHB tolerance may be mediated by alterations in central nervous system pharmacokinetics or neurotransmitter response. Rats were dosed daily with GHB (548 mg/kg s.c. q.d. for 5 days), and sleep time was measured as an index of behavioral tolerance. Plasma and brain GHB pharmacokinetics on days 1 and 5 were monitored using blood and microdialysis sampling. Extracellular (ECF) striatal dopamine levels were measured by microdialysis as a pharmacodynamic endpoint of tolerance. Pharmacokinetic (PK)/pharmacodynamic (PD) modeling was performed to describe the plasma and brain disposition using an indirect response model with inhibition of dopamine synthesis rate to describe the pharmacodynamic response. GHB plasma and brain ECF concentration versus time profiles following acute or chronic exposure were not significantly different. GHB sedative/hypnotic tolerance was observed by day 5. Acute GHB administration resulted in a decrease in striatal ECF dopamine (DA) levels compared with baseline levels. GHB tolerance was reflected by a 60% decrease in dopamine area under the curve (effect and baseline): acute, 10.1 +/- 15.3% basal DA/min/10(-3) versus chronic, 4.73 +/- 1.49% basal DA/min/10(-3) (p < 0.05, n = 5; unpaired Student's t test). The PK/PD model revealed an increase in the IC50 following chronic exposure indicating decreased dopaminergic sensitivity toward the inhibitory effects of GHB. Our findings indicate that GHB pharmacokinetics do not contribute to behavioral tolerance; however, changes in neurotransmitter responsiveness may suggest specific neurochemical pathways involved in the development and expression of tolerance. PMID- 17138863 TI - 5-Hydroxytryptamine2C receptor contribution to m-chlorophenylpiperazine and N methyl-beta-carboline-3-carboxamide-induced anxiety-like behavior and limbic brain activation. AB - Activation of 5-hydroxytryptamine2C (5-HT(2C)) receptors by the 5-HT(2) receptor agonist m-chlorophenylpiperazine (m-CPP) elicits anxiety in humans and anxiety like behavior in animals. We compared the effects of m-CPP with the anxiogenic GABA(A) receptor inverse agonist N-methyl-beta-carboline-3-carboxamide (FG-7142) on both anxiety-like behavior and regional brain activation using functional magnetic resonance imaging (fMRI) in the rat. We also determined whether the selective 5-HT(2C) receptor antagonist SB 242084 [6-chloro-2,3-dihydro-5-methyl-N [6-[(2-methyl-3-pyridinyl)oxy]-3-pyridinyl]-1H-indole-1-carboxyamide dihydrochloride] would blunt m-CPP or FG-7142-induced neuronal activation. Both m CPP (3 mg/kg i.p.) and FG-7142 (10 mg/kg i.p.) elicited anxiety-like behavior when measured in the social interaction test, and pretreatment with SB 242084 (1 mg/kg i.p.) completely blocked the behavioral effects of both anxiogenic drugs. Regional brain activation in vivo in response to anxiogenic drug challenge was determined by blood oxygen level-dependent (BOLD) fMRI using a powerful 9.4T magnet. Region of interest analyses revealed that m-CPP and FG-7142 significantly increased BOLD signals in brain regions that have been linked to anxiety, including the amygdala, dorsal hippocampus, and medial hypothalamus. These BOLD signal increases were blocked by pretreatment with SB 242084. In contrast, injection of m-CPP and FG-7142 resulted in BOLD signal decreases in the medial prefrontal cortex that were not blocked by SB 242084. In conclusion, the brain activation signals produced by anxiogenic doses of both m-CPP and FG-7142 are mediated at least partially by the 5-HT(2C) receptor, indicating that this receptor is a key component in anxiogenic neural circuitry. PMID- 17138864 TI - Cell cycle arrest by the isoprenoids perillyl alcohol, geraniol, and farnesol is mediated by p21(Cip1) and p27(Kip1) in human pancreatic adenocarcinoma cells. AB - Pancreatic cancer, the fourth leading cause of cancer-associated mortality in the United States, usually presents in an advanced stage and is generally refractory to chemotherapy. As such, there is a great need for novel therapies for this disease. The naturally derived isoprenoids perillyl alcohol, farnesol, and geraniol have chemotherapeutic potential in pancreatic and other tumor types. However, their mechanisms of action in these systems are not completely defined. In this study, we investigated isoprenoid effects on the cell cycle and observed a similar antiproliferative mechanism of action among the three compounds. First, when given in combination, the isoprenoids exhibited an additive antiproliferative effect against MIA PaCa-2 human pancreatic cancer cells. Furthermore, all three compounds induced a G(0)/G(1) cell cycle arrest that coincided with an increase in the expression of the cyclin kinase inhibitor proteins p21(Cip1) and p27(Kip1) and a reduction in cyclin A, cyclin B1, and cyclin-dependent kinase (Cdk) 2 protein levels. Immunoprecipitation studies demonstrated increased association of both p21(Cip1) and p27(Kip1) with Cdk2 as well as diminished Cdk2 kinase activity after isoprenoid exposure, indicating a cell cycle-inhibitory role for p21(Cip1) and p27(Kip1) in pancreatic adenocarcinoma cells. When siRNA was used to inhibit expression of p21(Cip1) and p27(Kip1) proteins in MIA PaCa-2 cells, conditional resistance to all three isoprenoid compounds was evident. Given similar findings in this cell line and in BxPC-3 human pancreatic adenocarcinoma cells, we conclude that the chemotherapeutic isoprenoid compounds perillyl alcohol, farnesol, and geraniol invoke a p21(Cip1)- and p27(Kip1)-dependent antiproliferative mechanism in human pancreatic adenocarcinoma cells. PMID- 17138866 TI - Homoploid hybrid speciation in an extreme habitat. AB - According to theory, homoploid hybrid speciation, which is hybrid speciation without a change in chromosome number, is facilitated by adaptation to a novel or extreme habitat. Using molecular and ecological data, we found that the alpine adapted butterflies in the genus Lycaeides are the product of hybrid speciation. The alpine populations possess a mosaic genome derived from both L. melissa and L. idas and are differentiated from and younger than their putative parental species. As predicted, adaptive traits may allow for persistence in the environmentally extreme alpine habitat and reproductively isolate these populations from their parental species. PMID- 17138865 TI - Anti-inflammatory effects of 4-phenyl-3-butenoic acid and 5-(acetylamino)-4-oxo-6 phenyl-2-hexenoic acid methyl ester, potential inhibitors of neuropeptide bioactivation. AB - Substance P (SP) and calcitonin gene-related peptide (CGRP) are well established mediators of inflammation. Therefore, inhibition of the biosynthesis of these neuropeptides is an attractive potential strategy for pharmacological intervention against a number of inflammatory diseases. The final step in the biosynthesis of SP and CGRP is the conversion of their glycine-extended precursors to the active amidated peptide, and this process is catalyzed by sequential action of the enzymes peptidylglycine alpha-monooxygenase (PAM) and peptidylamidoglycolate lyase. We have demonstrated previously that 4-phenyl-3 butenoic acid (PBA) is a PAM inhibitor, and we have also shown that in vivo inhibition of serum PAM by PBA correlates with this compound's ability to inhibit carrageenan-induced edema in the rat. Here we demonstrate the ability of PBA to inhibit all three phases of adjuvant-induced polyarthritis (AIP) in rats; this represents the first time that an amidation inhibitor has been shown to be active in a model of chronic inflammation. We recently introduced 5-(acetylamino)-4-oxo 6-phenyl-2-hexenoic acid (AOPHA) as one of a new series of mechanism-based amidation inhibitors. We now report for the first time that AOPHA and its methyl ester (AOPHA-Me) are active inhibitors of serum PAM in vivo, and we show that AOPHA-Me correspondingly inhibits carrageenan-induced edema in rats in a dose dependent manner. Neither PBA nor AOPHA-Me exhibits significant cyclooxygenase (COX) inhibition in vitro; thus, the anti-inflammatory activities of PBA and AOPHA-Me are apparently not a consequence of COX inhibition. We discuss possible pharmacological mechanisms that may account for the activities of these new anti inflammatory compounds. PMID- 17138867 TI - Slow earthquakes coincident with episodic tremors and slow slip events. AB - We report on the very-low-frequency earthquakes occurring in the transition zone of the subducting plate interface along the Nankai subduction zone in southwest Japan. Seismic waves generated by very-low-frequency earthquakes with seismic moment magnitudes of 3.1 to 3.5 predominantly show a long period of about 20 seconds. The seismicity of very-low-frequency earthquakes accompanies and migrates with the activity of deep low-frequency tremors and slow slip events. The coincidence of these three phenomena improves the detection and characterization of slow earthquakes, which are thought to increase the stress on updip megathrust earthquake rupture zones. PMID- 17138868 TI - P[acman]: a BAC transgenic platform for targeted insertion of large DNA fragments in D. melanogaster. AB - We describe a transgenesis platform for Drosophila melanogaster that integrates three recently developed technologies: a conditionally amplifiable bacterial artificial chromosome (BAC), recombineering, and bacteriophage PhiC31-mediated transgenesis. The BAC is maintained at low copy number, facilitating plasmid maintenance and recombineering, but is induced to high copy number for plasmid isolation. Recombineering allows gap repair and mutagenesis in bacteria. Gap repair efficiently retrieves DNA fragments up to 133 kilobases long from P1 or BAC clones. PhiC31-mediated transgenesis integrates these large DNA fragments at specific sites in the genome, allowing the rescue of lethal mutations in the corresponding genes. This transgenesis platform should greatly facilitate structure/function analyses of most Drosophila genes. PMID- 17138869 TI - Spectropolarimetric diagnostics of thermonuclear supernova explosions. AB - Even at extragalactic distances, the shape of supernova ejecta can be effectively diagnosed by spectropolarimetry. We present results for 17 type Ia supernovae that allow a statistical study of the correlation among the geometric structures and other observable parameters of type Ia supernovae. These observations suggest that type Ia supernova ejecta typically consist of a smooth, central, iron-rich core and an outer layer with chemical asymmetries. The degree of this peripheral asphericity is correlated with the light-curve decline rate of type Ia supernovae. These results lend strong support to delayed-detonation models of type Ia supernovae. PMID- 17138870 TI - Responding to fraud. PMID- 17138871 TI - Scientific conduct. China's fraud buster hit by libel judgments; defenders rally round. PMID- 17138872 TI - Developmental biology. Fraud investigation clouds paper on early cell fate. PMID- 17138873 TI - Medicine. Squelching progesterone's signal may prevent breast cancer. PMID- 17138874 TI - Molecular biology. Three methods add up to one new way to genetically engineer fruit flies. PMID- 17138875 TI - Drug research. WHO panel weighs radical ideas. PMID- 17138876 TI - Science education. Doing more with less. PMID- 17138877 TI - Astrophysics. Burst-hunter's rich data harvest yields a cosmic enigma. PMID- 17138878 TI - Global health. South Africa bolsters HIV/AIDS plan, but obstacles remain. PMID- 17138879 TI - Wildlife conservation. The saola's last stand. PMID- 17138880 TI - Balancing communication and safety. PMID- 17138881 TI - Glossing over the complexity of water. PMID- 17138882 TI - Mitochondrial DNA and population size. PMID- 17138883 TI - Comment on "Population size does not influence mitochondrial genetic diversity in animals". AB - Bazin et al. (Reports, 28 April, 2006, p. 570) found no relationship between mitochondrial DNA (mtDNA) diversity and population size when comparing across large groups of animals. We show empirically that species with smaller populations, as represented by eutherian mammals, exhibit a positive correlation between mtDNA and allozyme variation, suggesting that mtDNA diversity may correlate with population size in these animals. PMID- 17138884 TI - Science and law. When patents threaten science. PMID- 17138885 TI - Developmental biology. The Turing model comes of molecular age. PMID- 17138886 TI - Astronomy. Variable high-energy gamma rays from the elliptical galaxy M87. PMID- 17138887 TI - Atmosphere. When dry air is too humid. PMID- 17138888 TI - Genetics. Delivering new disease genes. PMID- 17138889 TI - Retrospective: Edward I. Stiefel (1942-2006). PMID- 17138891 TI - Brassinosteroid signaling: a paradigm for steroid hormone signaling from the cell surface. AB - Plants use the coordinated action of several small-molecule hormones to grow and develop optimally in response to a changing environment. Among these hormones are the brassinosteroids (BRs), the polyhydroxylated steroid hormones of plants. BRs bind a small family of leucine-rich repeat receptor kinases at the cell surface, thereby initiating an intracellular signal transduction cascade that results in the altered expression of hundreds of genes. PMID- 17138892 TI - G protein signaling in yeast: new components, new connections, new compartments. AB - Signaling by cell surface receptors and heterotrimeric guanine nucleotide-binding proteins (G proteins) is one of the most exhaustively studied processes in the cell but remains a major focus of molecular pharmacology research. The pheromone response system in yeast (see the Connections Map at Science's Signal Transduction Knowledge Environment) has provided numerous major advances in our understanding of G protein signaling and regulation. However, the basic features of this prototypical pathway have remained largely unchanged since the mid-1990s. New tools available in yeast are beginning to uncover new pathway components and interactions and have revealed signaling in unexpected locations within the cell. PMID- 17138893 TI - Notch, a universal arbiter of cell fate decisions. AB - Members of the Notch family of receptors act as membrane-tethered transcription factors that are tightly associated with binary cell fate decisions. Notch signaling acts as a molecular gate that allows cells to adopt or forfeit a particular fate. Interaction of Notch with ligands triggers a sequence of proteolytic cleavages that release the intracellular domain to the nucleus; this mechanism is a target of therapies for leukemias associated with Notch activation. Although the molecular mechanism of Notch activation is well characterized, further analysis in an appropriate cellular context will provide new insight into Notch signaling. PMID- 17138894 TI - Old-growth forests can accumulate carbon in soils. AB - Old-growth forests have traditionally been considered negligible as carbon sinks because carbon uptake has been thought to be balanced by respiration. We show that the top 20-centimeter soil layer in preserved old-growth forests in southern China accumulated atmospheric carbon at an unexpectedly high average rate of 0.61 megagrams of carbon hectare-1 year-1 from 1979 to 2003. This study suggests that the carbon cycle processes in the belowground system of these forests are changing in response to the changing environment. The result directly challenges the prevailing belief in ecosystem ecology regarding carbon budget in old-growth forests and supports the establishment of a new, nonequilibrium conceptual framework to study soil carbon dynamics. PMID- 17138895 TI - Solid-state qubits with current-controlled coupling. AB - The ability to switch the coupling between quantum bits (qubits) on and off is essential for implementing many quantum-computing algorithms. We demonstrated such control with two flux qubits coupled together through their mutual inductances and through the dc superconducting quantum interference device (SQUID) that reads out their magnetic flux states. A bias current applied to the SQUID in the zero-voltage state induced a change in the dynamic inductance, reducing the coupling energy controllably to zero and reversing its sign. PMID- 17138896 TI - Optical atomic coherence at the 1-second time scale. AB - Highest-resolution laser spectroscopy has generally been limited to single trapped ion systems because of the rapid decoherence that plagues neutral atom ensembles. Precision spectroscopy of ultracold neutral atoms confined in a trapping potential now shows superior optical coherence without any deleterious effects from motional degrees of freedom, revealing optical resonance linewidths at the hertz level with a good signal-to-noise ratio. The resonance quality factor of 2.4 x 10(14) is the highest ever recovered in any form of coherent spectroscopy. The spectral resolution permits direct observation of the breaking of nuclear spin degeneracy for the 1S0 and 3P0 optical clock states of 87Sr under a small magnetic bias field. This optical approach for excitation of nuclear spin states allows an accurate measurement of the differential Lande g factor between 1S0 and 3P0. The optical atomic coherence demonstrated for collective excitation of a large number of atoms will have a strong impact on quantum measurement and precision frequency metrology. PMID- 17138897 TI - Macroscopic hierarchical surface patterning of porphyrin trimers via self assembly and dewetting. AB - The use of bottom-up approaches to construct patterned surfaces for technological applications is appealing, but to date is applicable to only relatively small areas (approximately 10 square micrometers). We constructed highly periodic patterns at macroscopic length scales, in the range of square millimeters, by combining self-assembly of disk-like porphyrin dyes with physical dewetting phenomena. The patterns consisted of equidistant 5-nanometer-wide lines spaced 0.5 to 1 micrometers apart, forming single porphyrin stacks containing millions of molecules, and were formed spontaneously upon drop-casting a solution of the molecules onto a mica surface. On glass, thicker lines are formed, which can be used to align liquid crystals in large domains of square millimeter size. PMID- 17138898 TI - Organic globules in the Tagish Lake meteorite: remnants of the protosolar disk. AB - Coordinated transmission electron microscopy and isotopic measurements of organic globules in the Tagish Lake meteorite shows that they have elevated ratios of nitrogen-15 to nitrogen-14 (1.2 to 2 times terrestrial) and of deuterium to hydrogen (2.5 to 9 times terrestrial). These isotopic anomalies are indicative of mass fractionation during chemical reactions at extremely low temperatures (10 to 20 kelvin), characteristic of cold molecular clouds and the outer protosolar disk. The globules probably originated as organic ice coatings on preexisting grains that were photochemically processed into refractory organic matter. The globules resemble cometary carbon, hydrogen, oxygen, and nitrogen (CHON) particles, suggesting that such grains were important constituents of the solar system starting materials. PMID- 17138899 TI - Increasing trend of extreme rain events over India in a warming environment. AB - Against a backdrop of rising global surface temperature, the stability of the Indian monsoon rainfall over the past century has been a puzzle. By using a daily rainfall data set, we show (i) significant rising trends in the frequency and the magnitude of extreme rain events and (ii) a significant decreasing trend in the frequency of moderate events over central India during the monsoon seasons from 1951 to 2000. The seasonal mean rainfall does not show a significant trend, because the contribution from increasing heavy events is offset by decreasing moderate events. A substantial increase in hazards related to heavy rain is expected over central India in the future. PMID- 17138900 TI - Male fertility and sex ratio at birth in red deer. AB - Efforts to test sex ratio theory have focused mostly on females. However, when males possess traits that could enhance the reproductive success of sons, males would also benefit from the manipulation of the offspring sex ratio. We tested the prediction that more-fertile red deer males produce more sons. Our findings reveal that male fertility is positively related to the proportion of male offspring. We also show that there is a positive correlation between the percentage of morphologically normal spermatozoa (a main determinant of male fertility) and the proportion of male offspring. Thus, males may contribute significantly to biases in sex ratio at birth among mammals, creating the potential for conflicts of interest between males and females. PMID- 17138901 TI - Microfluidic digital PCR enables multigene analysis of individual environmental bacteria. AB - Gene inventory and metagenomic techniques have allowed rapid exploration of bacterial diversity and the potential physiologies present within microbial communities. However, it remains nontrivial to discover the identities of environmental bacteria carrying two or more genes of interest. We have used microfluidic digital polymerase chain reaction (PCR) to amplify and analyze multiple, different genes obtained from single bacterial cells harvested from nature. A gene encoding a key enzyme involved in the mutualistic symbiosis occurring between termites and their gut microbiota was used as an experimental hook to discover the previously unknown ribosomal RNA-based species identity of several symbionts. The ability to systematically identify bacteria carrying a particular gene and to link any two or more genes of interest to single species residing in complex ecosystems opens up new opportunities for research on the environment. PMID- 17138902 TI - Prevention of Brca1-mediated mammary tumorigenesis in mice by a progesterone antagonist. AB - Women with mutations in the breast cancer susceptibility gene BRCA1 are predisposed to breast and ovarian cancers. Why the BRCA1 protein suppresses tumor development specifically in ovarian hormone-sensitive tissues remains unclear. We demonstrate that mammary glands of nulliparous Brca1/p53-deficient mice accumulate lateral branches and undergo extensive alveologenesis, a phenotype that occurs only during pregnancy in wild-type mice. Progesterone receptors, but not estrogen receptors, are overexpressed in the mutant mammary epithelial cells because of a defect in their degradation by the proteasome pathway. Treatment of Brca1/p53-deficient mice with the progesterone antagonist mifepristone (RU 486) prevented mammary tumorigenesis. These findings reveal a tissue-specific function for the BRCA1 protein and raise the possibility that antiprogesterone treatment may be useful for breast cancer prevention in individuals with BRCA1 mutations. PMID- 17138903 TI - Immigration and mental health: diverse findings in Asian, black, and Latino populations. PMID- 17138904 TI - Is educational inequality protective? PMID- 17138905 TI - Use of mental health-related services among immigrant and US-born Asian Americans: results from the National Latino and Asian American Study. AB - OBJECTIVES: We examined rates of mental health-related service use (i.e., any, general medical, and specialty mental health services) as well as subjective satisfaction with and perceived helpfulness of care in a national sample of Asian Americans, with a particular focus on immigration-related factors. METHODS: Data were derived from the National Latino and Asian American Study (2002-2003). RESULTS: About 8.6% of the total sample (n=2095) sought any mental health-related services; 34.1% of individuals who had a probable diagnosis sought any services. Rates of mental health-related service use, subjective satisfaction, and perceived helpfulness varied by birthplace and by generation. US-born Asian Americans demonstrated higher rates of service use than did their immigrant counterparts. Third-generation or later individuals who had a probable diagnosis had high (62.6%) rates of service use in the previous 12 months. CONCLUSIONS: Asian Americans demonstrated lower rates of any type of mental health-related service use than did the general population, although there are important exceptions to this pattern according to nativity status and generation status. Our results underscore the importance of immigration-related factors in understanding service use among Asian Americans. PMID- 17138906 TI - Explaining recent declines in adolescent pregnancy in the United States: the contribution of abstinence and improved contraceptive use. AB - OBJECTIVES: We explored the relative contributions of declining sexual activity and improved contraceptive use to the recent decline in adolescent pregnancy rates in the United States. METHODS: We used data from 1995 and 2002 for women 15 to 19 years of age to develop 2 indexes: the contraceptive risk index, summarizing the overall effectiveness of contraceptive use among sexually active adolescents (including nonuse), and the overall pregnancy risk index, calculated according to the contraceptive risk index score and the percentage of individuals reporting sexual activity. RESULTS: The contraceptive risk index declined 34% overall and 46% among adolescents aged 15 to 17 years. Improvements in contraceptive use included increases in the use of condoms, birth control pills, withdrawal, and multiple methods and a decline in nonuse. The overall pregnancy risk index declined 38%, with 86% of the decline attributable to improved contraceptive use. Among adolescents aged 15 to 17 years, 77% of the decline in pregnancy risk was attributable to improved contraceptive use. CONCLUSIONS: The decline in US adolescent pregnancy rates appears to be following the patterns observed in other developed countries, where improved contraceptive use has been the primary determinant of declining rates. PMID- 17138907 TI - Use of mental health services and subjective satisfaction with treatment among Black Caribbean immigrants: results from the National Survey of American Life. AB - OBJECTIVES: We examined the use rates and correlates of formal psychiatric services among the US-born and immigrant Caribbean Black population. METHODS: We compared overall mental health service use in samples of Caribbean Blacks and African Americans and examined the within-sample ethnic variation among Caribbean Blacks, including for ethnic origin (Spanish Caribbean, Haiti, and English Caribbean), nativity status (those born in or outside the United States), number of years spent living in the United States, age at the time of immigration, and generational status. RESULTS: African Americans and Caribbean Blacks used formal mental health care services at relatively low rates. Among Caribbean Blacks, generational status and nativity showed the greatest effects on rates of reported use, satisfaction, and perceived helpfulness. Of those study participants who met the criteria for disorders as defined by the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, about one third used formal mental health care services. The US-born subjects were more likely to receive care than were first generation immigrants. CONCLUSIONS: Our study underscores the importance of ethnicity, immigration, and migration-related factors, within racial categorization, as it pertains to the use of mental health services in the United States. Our findings suggest that timing of migration and generational status of Caribbean Black immigrants and ancestry groups contribute to important differences in rates and sources of use, relative satisfaction, and perception of helpfulness, with regard to formal mental health services. PMID- 17138908 TI - Immigration-related factors and mental disorders among Asian Americans. AB - OBJECTIVES: We examined lifetime and 12-month rates of any depressive, anxiety, and substance abuse disorders in a national sample of Asian Americans. We focused on factors related to nativity and immigration as possible correlates of mental disorders. METHODS: Data were derived from the National Latino and Asian American Study, the first national epidemiological survey of Asian Americans in the United States. RESULTS: The relationships between immigration-related factors and mental disorders were different for men and women. Among women, nativity was strongly associated with lifetime disorders, with immigrant women having lower rates of most disorders compared with US-born women. Conversely, English proficiency was associated with mental disorders for Asian men. Asian men who spoke English proficiently generally had lower rates of lifetime and 12-month disorders compared with nonproficient speakers. CONCLUSIONS: For Asian Americans, immigration-related factors were associated with mental disorders, but in different ways for men and women. Future studies will need to examine gender as an important factor in specifying the association between immigration and mental health. PMID- 17138909 TI - The mental health of Black Caribbean immigrants: results from the National Survey of American Life. AB - OBJECTIVES: We examined the prevalence of psychiatric disorders among Black Caribbean immigrant ("Caribbean Black") and African American populations and the correlates of psychiatric disorders among the Caribbean Black population. METHODS: We conducted descriptive and age-adjusted analyses of the data from the National Survey of American Life--an in-person household mental health survey of noninstitutionalized US Blacks. We assessed psychiatric disorders as defined by Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria with the Composite International Diagnostic Interview. RESULTS: Compared with African American men, Caribbean Black men had higher risks for 12-month rates of psychiatric disorders. Caribbean Black women had lower odds for 12-month and lifetime psychiatric disorders compared with African American women. Risks varied by ethnicity, immigration history, and generation status within the Caribbean sample. First-generation Caribbean Blacks had lower rates of psychiatric disorders compared with second- or third-generation Caribbean Blacks, and, compared with first-generation Caribbean Blacks, third-generation Caribbean Blacks had markedly elevated rates of psychiatric disorders. CONCLUSIONS: Mental health risks were associated with ethnic diversity within the US Black population. Increased exposure to minority status in the United States was associated with higher risks for psychiatric disorders among Black Caribbean immigrants, which possibly reflects increased societal stress and downward social mobility associated with being Black in America. PMID- 17138910 TI - Prevalence of psychiatric disorders across Latino subgroups in the United States. AB - OBJECTIVES: We examined the prevalence of depressive, anxiety, and substance use disorders among Latinos residing in the United States. METHODS: We used data from the National Latino and Asian American Study, which included a nationally representative sample of Latinos. We calculated weighted prevalence rates of lifetime and past-year psychiatric disorders across different sociodemographic, ethnic, and immigration groups. RESULTS: Lifetime psychiatric disorder prevalence estimates were 28.1% for men and 30.2% for women. Puerto Ricans had the highest overall prevalence rate among the Latino ethnic groups assessed. Increased rates of psychiatric disorders were observed among US-born, English-language proficient, and third-generation Latinos. CONCLUSIONS: Our results provide important information about potential correlates of psychiatric problems among Latinos that can inform clinical practice and guide program development. Stressors associated with cultural transmutation may exert particular pressure on Latino men. Continued attention to environmental influences, especially among third-generation Latinos, is an important area for substance abuse program development. PMID- 17138911 TI - Correlates of past-year mental health service use among Latinos: results from the National Latino and Asian American Study. AB - OBJECTIVES: We examined correlates and rates of past-year mental health service use in a national sample of Latinos residing in the United States. METHODS: We used data from the National Latino and Asian American Study, a national epidemiological household survey of Latinos. RESULTS: Cultural factors such as nativity, language, age at migration, years of residence in the United States, and generational status were associated with whether or not Latinos had used mental health services. However, when the analysis was stratified according to past-year psychiatric diagnoses, these associations held only among those who did not fulfill criteria for any of the psychiatric disorders assessed. Rates of mental health service use among those who did not fulfill diagnostic criteria were higher among Puerto Ricans and US-born Latinos than among non-Puerto Ricans and foreign-born Latinos. CONCLUSIONS: Rates of mental health service use among Latinos appear to have increased substantially over the past decade relative to rates reported in the 1990s. Cultural and immigration characteristics should be considered in matching mental health services to Latinos who need preventive services or who are symptomatic but do not fulfill psychiatric disorder criteria. PMID- 17138912 TI - Promises and challenges of faith-based AIDS care and support in Mozambique. AB - OBJECTIVES: We sought to examine the role of religious organizations in the provision of HIV/AIDS-related assistance in Africa. METHODS: We used data collected from Christian religious organizations in southern Mozambique. Bivariate comparisons and logistic regression analysis of survey data were performed. We conducted an analysis of the qualitative data to complement the quantitative results. RESULTS: Our analysis revealed little involvement of religious organizations in provision of assistance. Most assistance was decentralized and consisted of psychological support and some personal care and household help. Material or financial help was rare. Assistance to nonmembers of congregations was reported more often than to members. Members of larger and better-secularly connected congregations were more likely to report assistance than were members of smaller and less-secularly engaged ones. Assistance was reported more in cities than in rural areas. Women were more likely than men to report providing assistance to congregation members, and the reverse was true for assistance provided to nonmembers. The cooperation of religious organizations in provision of assistance was hindered by financial constraints and institutional rivalry. CONCLUSIONS: Policy efforts to involve religious organizations in provision of HIV/AIDS-related assistance should take into account that organization's resources, institutional goals, and social characteristics. PMID- 17138913 TI - Informed consent: an international researchers' perspective. AB - We reported 164 researchers' recommendations for information that should be included in the informed consent process. These recommendations were obtained during training workshops conducted in Africa, Europe, and the United States. The 8 elements of informed consent of the US Code of Federal Regulations were used to identify 95 items of information ("points"), most related to benefits and research description. Limited consensus was found among the 3 workshops: of the 95 points, only 27 (28%) were identified as useful by all groups. These points serve as a springboard for identifying information applicable in different geographic areas and indicate the need for involving a variety of individuals and stakeholders, with different research and cultural perspectives, in the development of informed consent, particularly for research undertaken in international settings. PMID- 17138914 TI - The impact of cost on the availability of fruits and vegetables in the homes of schoolchildren in Birmingham, Alabama. AB - OBJECTIVES: Fruit and vegetable cost is a presumed barrier to intake. We sought to determine whether fruit and vegetable cost and consumers' race and income would predict availability of fruits and vegetables in homes of schoolchildren in the Birmingham, Ala, area. METHODS: Data on availability of 27 fruit and vegetable items were obtained from homes of 1355 children (32% African American) in the Birmingham area. Fruit and vegetable costs were obtained from the US Department of Agriculture. We used discrete choice analysis with the dependent variable represented as presence or absence of the fruit or vegetable item. Explanatory variables included fruit and vegetable price per serving; child's gender, race, and age; and parent's body mass index and income. RESULTS: Higher cost was inversely related to fruit and vegetable availability. Higher income, African American race, and female gender were positively related to availability. Cost per serving was stratified into 3 categories-low, medium, and high. Relative to low-cost items, only high-cost items decreased the odds of availability significantly. CONCLUSIONS: Fruit and vegetable cost does impact availability and has the greatest impact for high-cost items. Although cost was inversely related to availability, African Americans reported higher fruit and vegetable availability than Whites. Additional studies are needed to determine whether food items of lower nutritive value and comparable cost impact availability. PMID- 17138915 TI - Transnational tobacco company influence on tax policy during privatization of a state monopoly: British American Tobacco and Uzbekistan. AB - OBJECTIVES: The International Monetary Fund encourages privatization of state owned tobacco industries. Privatization tends to lower cigarette prices, which encourages consumption. This could be countered with effective tax policies. We explored how investment by British American Tobacco (BAT) influenced tax policy in Uzbekistan during privatization there. METHODS: We obtained internal documents from BAT and analyzed them using a hermeneutic process to create a chronology of events. RESULTS: BAT thoroughly redesigned the tobacco taxation system in Uzbekistan. It secured (1) a reduction of approximately 50% in the excise tax on cigarettes, (2) an excise system to benefit its brands and disadvantage those of its competitors (particularly Philip Morris), and (3) a tax stamp system from which it hoped to be exempted, because this would likely facilitate its established practice of cigarette smuggling and further its competitive advantage.. CONCLUSIONS: Privatization can endanger effective tobacco excise policies. The International Monetary Fund should review its approach to privatization and differentiate the privatization of an industry whose product kills from privatization of other industries. PMID- 17138917 TI - A statewide public health approach to improving organ donation: the Massachusetts Organ Donation Initiative. AB - Despite the growing disparity between organ supply and demand in the United States, few initiatives have attempted to close the gap through systematic population-based public health endeavors. We examined the evolution, implementation, and outcomes of the Massachusetts Organ Donation Initiative, a statewide effort that included a unique partnership among organ procurement organizations, major teaching hospitals, and the state's department of public health. Lessons from this initiative have contributed to growing national efforts for increasing organ supply and have provided insights for addressing this continuing public health challenge. PMID- 17138916 TI - Assessment of HIV testing of urban injection drug users: implications for expansion of HIV testing and prevention efforts. AB - OBJECTIVES: We sought to determine the extent of HIV testing among urban injection drug users (IDUs) to assess whether an expansion of targeted testing programs would be consistent with national goals to identify previously undetected infections. METHODS: IDUs in 5 US cities (Oakland, Calif; Chicago, Ill; Hartford and New Haven, Conn; and Springfield, Mass) were recruited either by chain referral or time-location sampling. The IDUs were questioned about HIV testing, and factors associated with HIV testing were analyzed. RESULTS: Ninety three percent of 1543 IDUs had been tested. Among those tested but who did not report having been told that they were HIV seropositive, 90% had been tested within the past 3 years. Women and syringe-exchange customers were more likely to have been tested ever and in the recent past. We estimated the number of undetected infections among urban IDUs in the United States to be less than 40000. CONCLUSIONS: Testing for HIV has reached the vast majority of IDUs through the current options. Expending scarce prevention money to expand testing of IDUs is unlikely to be productive. Instead, resources should be used for proven HIV prevention strategies including syringe exchange, drug treatment, and secondary prevention for those who are HIV positive. PMID- 17138918 TI - Characteristics of persons with heterosexually acquired HIV infection, United States 1999-2004. AB - OBJECTIVES: In the United States a growing proportion of cases of heterosexually acquired HIV infections occur in women and in persons of color. We analyzed the association between race/ethnicity, whether diagnoses of HIV infection and AIDS were made concurrently, and the survival after diagnosis of heterosexually acquired AIDS. METHODS: We used data from 29 states that report confidential name based HIV/AIDS cases to the Centers for Disease Control and Prevention to calculate estimated annual percentage change in the number of actual diagnoses and analyzed the association between race/ethnicity and concurrent diagnoses of HIV and AIDS. We adjusted for reporting delays and absence of information about HIV risk factors. RESULTS: During 1999 to 2004, diagnoses of heterosexually acquired HIV were made for 52 569 persons in 29 states; 33 554 (64%) were women. Among men and women, 38 470 (73%) were non-Hispanic Black; 7761 (15%), non Hispanic White; and 5383 (10%), Hispanic. The number of persons with heterosexually acquired HIV significantly increased: 6.1% among Hispanic men (95% confidence interval=2.7, 9.7) and 4.5% among Hispanic women (95% confidence interval=1.8, 7.3). The number significantly decreased (-2.9%) among non-Hispanic Black men. Concurrent HIV and AIDS diagnoses were slightly more common for non Hispanic Whites (23%) and Hispanics (23%) than for non-Hispanic Blacks (20%). CONCLUSIONS: To decrease the incidence of heterosexually acquired HIV infections, prevention and education programs should target all persons at risk, especially women, non-Hispanic Blacks, and Hispanics. PMID- 17138919 TI - Adolescent participation in preventive health behaviors, physical activity, and nutrition: differences across immigrant generations for Asians and Latinos compared with Whites. AB - OBJECTIVES: We investigated preventive health behaviors (bicycle helmet, seat belt, and sunscreen use), physical activity, television viewing or video game playing, and nutrition (fruit, vegetable, milk, and soda consumption) among Asian and Latino adolescents living in the United States; assessed trends across generations (first-, second-, and third-generation immigrants or later); and compared each generation with White adolescents. METHODS: We used data from 5801 adolescents aged 12 to 17 years in the representative 2001 California Health Interview Survey. RESULTS: In multivariate analysis, first-generation Asians measured worse than Whites for preventive health behaviors (lower participation), physical activity (less activity), and television viewing or video game playing (more hours), but improved across generations. For these same behaviors, Latinos were similar to or worse than Whites, and generally showed no improvement across generations. First-generation Asians and Latinos had healthier diets than Whites (higher fruit and vegetable consumption, lower soda consumption). With succeeding generations, Asians' fruit, vegetable, and soda consumption remained stable, but Latinos' fruit and vegetable consumption decreased and their soda consumption increased, so that by the third generation Latinos' nutrition was poorer than Whites'. CONCLUSIONS: For the health behaviors we examined, Asian adolescents' health behaviors either improved with each generation or remained better than that of Whites. Latino adolescents demonstrated generally worse preventive health behaviors than did Whites and, in the case of nutrition, a worsening across generations. Targeted interventions may be needed to address behavioral disparities. PMID- 17138920 TI - Best-evidence interventions: findings from a systematic review of HIV behavioral interventions for US populations at high risk, 2000-2004. AB - OBJECTIVES: The Centers for Disease Control and Prevention's HIV/AIDS Prevention Research Synthesis Team conducted a systematic review of US-based HIV behavioral intervention research literature from 2000 through 2004 to identify interventions demonstrating best evidence of efficacy for reducing HIV risk. METHODS: Standard systematic review methods were used. Each eligible study was reviewed on the basis of Prevention Research Synthesis Team efficacy criteria that focused on 3 domains: study design, implementation and analysis, and strength of evidence. RESULTS: Eighteen interventions met the criteria for best evidence. Four targeted HIV-positive individuals. Of those targeting populations at risk for HIV, 4 targeted drug users, 6 targeted adults at risk because of heterosexual behaviors only, 2 targeted men who have sex with men, and 2 targeted youths at high risk. Eight interventions focused on women, and 13 had study samples with more than 50% minority participants. Significant intervention effects included increased condom use and reductions in unprotected sexual intercourse, number of sexual partners, injection drug use or needle sharing, and newly acquired sexually transmitted infections. CONCLUSIONS: Most of the best-evidence interventions are directly applicable for populations in greatest need of effective prevention programs; however, important gaps still exist. PMID- 17138921 TI - Sexual and drug behavior patterns and HIV and STD racial disparities: the need for new directions. AB - OBJECTIVES: We used nationally representative data to examine whether individuals' sexual and drug behavior patterns account for racial disparities in sexually transmitted disease (STD) and HIV prevalence. METHODS: Data were derived from wave III of the National Longitudinal Study of Adolescent Health. Participants were aged 18 to 26 years old; analyses were limited to non-Hispanic Blacks and Whites. Theory and cluster analyses yielded 16 unique behavior patterns. Bivariate analyses compared STD and HIV prevalences for each behavior pattern, by race. Logistic regression analyses examined within-pattern race effects before and after control for covariates. RESULTS: Unadjusted odds of STD and HIV infection were significantly higher among Blacks than among Whites for 11 of the risk behavior patterns assessed. Across behavior patterns, covariates had little effect on reducing race odds ratios. CONCLUSIONS: White young adults in the United States are at elevated STD and HIV risk when they engage in high-risk behaviors. Black young adults, however, are at high risk even when their behaviors are normative. Factors other than individual risk behaviors and covariates appear to account for racial disparities, indicating the need for population-level interventions. PMID- 17138922 TI - Ethics in public health research: a research protocol to evaluate the effectiveness of public-private partnerships as a means to improve health and welfare systems worldwide. AB - Public-private partnerships have become a common approach to health care problems worldwide. Many public-private partnerships were created during the late 1990s, but most were focused on specific diseases such as HIV/AIDS, tuberculosis, and malaria. Recently there has been enthusiasm for using public-private partnerships to improve the delivery of health and welfare services for a wider range of health problems, especially in developing countries. The success of public private partnerships in this context appears to be mixed, and few data are available to evaluate their effectiveness. This analysis provides an overview of the history of health-related public-private partnerships during the past 20 years and describes a research protocol commissioned by the World Health Organization to evaluate the effectiveness of public-private partnerships in a research context. PMID- 17138924 TI - Maternal obesity in early pregnancy and risk of spontaneous and elective preterm deliveries: a retrospective cohort study. AB - OBJECTIVES: We sought to determine the association between maternal body mass index and risk of preterm delivery. METHODS: We assessed 187,290 women in Scotland and estimated adjusted odds ratios for spontaneous and elective preterm deliveries among overweight, obese, and morbidly obese women relative to normal weight women. RESULTS: Among nulliparous women, the risk of requiring an elective preterm delivery increased with increasing BMI, whereas the risk of spontaneous preterm labor decreased. Morbidly obese nulliparous women were at increased risk of all-cause preterm deliveries, neonatal death, and delivery of an infant weighing less than 1000 g who survived to 1 year of age (a proxy for severe long term disability). By contrast, obesity and elective preterm delivery were only weakly associated among multiparous women. CONCLUSIONS: Obese nulliparous women are at increased risk of elective preterm deliveries. This in turn leads to an increased risk of perinatal mortality and is likely to lead to increased risks of long-term disability among surviving offspring. PMID- 17138923 TI - Going beyond "ABC" to include "GEM": critical reflections on progress in the HIV/AIDS epidemic. AB - A considerable number of studies have sought to identify what factors accounted for substantial reductions in HIV seroprevalence after several countries deployed "ABC" (abstinence, be faithful, condom use) strategies. After much public discourse and research on ABC success stories, the Joint United Nations Programme on HIV/AIDS 2004 epidemic report indicated that nearly 50% of infected people worldwide were women, up from 35% in 1985. In light of the feminization of HIV/AIDS, we critically assess the limitations of ABC strategies. We provide 3 additional prevention strategies that focus on gender relations, economics, and migration (GEM) and can speak to the new face of the epidemic. Pressing beyond ABC, GEM strategies provide the basis for a stronger central platform from which national efforts against HIV/AIDS can proceed to reduce transmission risks. PMID- 17138925 TI - Basic health, women's health, and mental health among internally displaced persons in Nyala Province, South Darfur, Sudan. AB - OBJECTIVES: We assessed basic health, women's health, and mental health among Sudanese internally displaced persons in South Darfur. METHODS: In January 2005, we surveyed 6 registered internally displaced persons camps in Nyala District. Using systematic random sampling, we surveyed 1293 households, interviewing 1 adult female per household (N=1274); respondents' households totaled 8643 members. We inquired about respondents' mental health, opinions on women's rights, and the health status of household members. RESULTS: A majority of respondents had access to rations, shelter, and water. Sixty-eight percent (861 of 1266) used no birth control, and 53% (614 of 1147) reported at least 1 unattended birth. Thirty percent (374 of 1238) shared spousal decisions on timing and spacing of children, and 49% (503 of 1027) reported the right to refuse sex. Eighty-four percent (1043 of 1240) were circumcised. The prevalence of major depression was 31% (390 of 1253). Women also expressed limited rights regarding marriage, movement, and access to health care. Eighty-eight percent (991 of 1121) supported equal educational opportunities for women. CONCLUSIONS: Humanitarian aid has relieved a significant burden of this displaced population's basic needs. However, mental and women's health needs remain largely unmet. The findings indicate a limitation of sexual and reproductive rights that may negatively affect health. PMID- 17138926 TI - Associations between family support, family intimacy, and neighborhood violence and physical activity in urban adolescent girls. AB - We examined the association between various dimensions of the family environment, including family intimacy and involvement in activities, family support for physical activity, and neighborhood violence (perceived and objective) and physical activity among urban, predominantly African American, ninth-grade girls in Baltimore, Md. Greater family intimacy (P = .05) and support (P = .01), but not neighborhood violence, was associated with physical activity. Family factors, including family intimacy and support, are potential targets in physical activity interventions for urban high-school girls. PMID- 17138927 TI - Prevalence of posttraumatic stress disorder in a coastal fishing village in Tamil Nadu, India, after the December 2004 tsunami. AB - Two months after the December 2004 tsunami in Tamil Nadu, India, we surveyed adults aged 18 years or older in a severely affected coastal village using structured interviews and the Harvard Trauma Questionnaire. The prevalence of posttraumatic stress disorder was 12.7% (95% confidence interval [CI]=9.4%, 17.1%), and odds of posttraumatic stress disorder were higher among individuals with no household incomes, women, and those injured during the tsunami. In addition to promoting feelings of safety, interventions aimed toward populations affected by the December 2004 tsunami need to focus on income-generating activities. Also, there is a need to target initiatives toward women and those individuals injured during the tsunami, given that these groups are more likely to experience posttraumatic stress disorder. PMID- 17138928 TI - Funding of North Carolina tobacco control programs through the Master Settlement Agreement. AB - Changing political and economic forces in 1 tobacco-dependent state, North Carolina, demonstrate how the interplay between these forces and public health priorities has shaped current allocation of Master Settlement Agreement funds. Allocation patterns demonstrate lawmakers' changing priorities in response to changes in the economic climate; some of the agreement's funds targeted to tobacco farmers appear to reflect objectives favored by tobacco manufacturers. Funds earmarked for health have underfunded youth tobacco prevention and tobacco control initiatives, and spending for tobacco farmers in North Carolina has not lived up to the rhetoric that accompanied the original agreement. We discuss the implications of these findings for future partnerships between public health advocates and workers as well as tobacco control strategies. PMID- 17138929 TI - Multilevel community-based intervention to increase access to sterile syringes among injection drug users through pharmacy sales in New York City. AB - OBJECTIVES: Research has indicated that there is minimal use of pharmacies among injection drug users (IDUs) in specific neighborhoods and among Black and Hispanic IDUs. We developed a community-based participatory research partnership to determine whether a multilevel intervention would increase sterile syringe access through a new policy allowing nonprescription syringe sales in pharmacies. METHODS: We targeted Harlem, NY (using the South Bronx for comparison), and disseminated informational material at community forums, pharmacist training programs, and counseling or outreach programs for IDUs. We compared cross sectional samples in 3 target populations (pre- and postintervention): community members (attitudes and opinions), pharmacists (opinions and practices), and IDUs (risk behaviors). RESULTS: Among community members (N = 1496) and pharmacists (N = 131), negative opinions of IDU syringe sales decreased in Harlem whereas there was either no change or an increase in negative opinions in the comparison community. Although pharmacy use by IDUs (N=728) increased in both communities, pharmacy use increased significantly among Black IDUs in Harlem, but not in the comparison community; syringe reuse significantly decreased in Harlem, but not in the comparison community. CONCLUSIONS: Targeting the individual and the social environment through a multilevel community-based intervention reduced high-risk behavior, particularly among Black IDUs. PMID- 17138930 TI - Impact of California's Proposition 36 on the drug treatment system: treatment capacity and displacement. AB - OBJECTIVES: California's Proposition 36 offers nonviolent drug offenders community-based treatment as an alternative to incarceration or probation without treatment. We examined how treatment capacity changed to accommodate Proposition 36 clients and whether displacement of other clients was an unintended consequence. METHODS: Treatment admissions were compared for the year before and 2 years after the law was enacted. Surveys of county administrators and treatment providers were conducted in Kern, Riverside, Sacramento, San Diego, and San Francisco counties. RESULTS: The number of Proposition 36 offenders admitted to treatment continued to increase in the state (approximately 32000 in Year 1 and 48000 in Year 2) and in the 5 counties; total treatment admissions stabilized in Year 2 after increasing in Year 1. Voluntary clients decreased by 8000 each year statewide, but the change varied across counties. One third of treatment providers reported decreased treatment availability for non-Proposition 36 clients in Year 2. CONCLUSION: Despite expanded treatment capacity (mostly in outpatient treatment), indirect evidence suggests that displacement of voluntary clients may have occurred in part because of the demand for treatment by Proposition 36 clients. PMID- 17138931 TI - Racial disparity in hypertensive disorders of pregnancy in New York State: a 10 year longitudinal population-based study. AB - OBJECTIVES: We studied trends of hypertensive disorders of pregnancy by residential socioeconomic status (SES) and racial/ethnic subgroups in New York State over a 10-year period. METHODS: We merged New York State discharge data for 2.5 million women hospitalized with delivery from 1993 through 2002 with 2000 US Census data. RESULTS: Rates of diagnoses for all hypertensive disorders combined and for preeclampsia individually were highest among Black women across all regions and neighborhood poverty levels. Although hospitalization rates for preeclampsia decreased over time for most groups, differences in rates between White and Black women increased over the 10-year period. The proportion of women living in poor areas remained relatively constant over the same period. Black and Hispanic women were more likely than White women to have a form of diabetes and were at higher risk of preeclampsia; preeclampsia rates were higher in these groups both with and without diabetes than in corresponding groups of White women. CONCLUSIONS: An increasing trend of racial/ethnic disparity in maternal hypertension rates occurred in New York State during the past decade. This trend was persistent after stratification according to SES and other risk factors. Additional research is needed to understand the factors contributing to this growing disparity. PMID- 17138932 TI - A national survey of tobacco cessation programs for youths. AB - OBJECTIVES: We collected data on a national sample of existing community-based tobacco cessation programs for youths to understand their prevalence and overall characteristics. METHODS: We employed a 2-stage sampling design with US counties as the first-stage probability sampling units. We then used snowball sampling in selected counties to identify administrators of tobacco cessation programs for youths. We collected data on cessation programs when programs were identified. RESULTS: We profiled 591 programs in 408 counties. Programs were more numerous in urban counties; fewer programs were found in low-income counties. State-level measures of smoking prevalence and tobacco control expenditures were not associated with program availability. Most programs were multisession, school based group programs serving 50 or fewer youths per year. Program content included cognitive-behavioral components found in adult programs along with content specific to adolescence. The median annual budget was 2000 dollars. Few programs (9%) reported only mandatory enrollment, 35% reported mixed mandatory and voluntary enrollment, and 56% reported only voluntary enrollment. CONCLUSIONS: There is considerable homogeneity among community-based tobacco cessation programs for youths. Programs are least prevalent in the types of communities for which national data show increases in youths' smoking prevalence. PMID- 17138933 TI - Serum amyloid P and cardiovascular disease in older men and women: results from the Cardiovascular Health Study. AB - OBJECTIVE: Serum amyloid P (SAP), a pentraxin like C-reactive protein (CRP), functions in innate immunity. However, associations of SAP with cardiovascular disease (CVD) are unknown. METHODS AND RESULTS: We examined these associations in the Cardiovascular Health Study using a case-cohort design. Nonexclusive case groups were incident angina (n=523), myocardial infarction (MI; n=308), stroke (n=323), and CVD death (n=288). 786 participants had no events. SAP was correlated with CRP, CVD risk factors (obesity, blood pressure, lipids), common and internal carotid wall thickness, and ankle-brachial index (all P<0.02). In Cox regression models adjusted for age, sex, and ethnicity, a standard deviation increase in SAP (9.8 mg/L) was associated with angina (hazard ratio; 95% confidence interval 1.3; 1.2 to 1.5) and MI (1.3; 1.1 to 1.5), but not stroke (1.1; 0.9 to 1.3) or CVD death (1.1; 0.9 to 1.3). Adding CRP to the models had no significant effect on associations. Adjusting for CVD risk factors slightly attenuated SAP associations with CVD events; however, associations with angina and MI remained significant. CONCLUSIONS: Although both are pentraxins, SAP and CRP may represent different facets of inflammation. The association of SAP with CVD in these older adults further supports the role of innate immunity in atherosclerosis. PMID- 17138934 TI - Adenovirus-mediated expression of tissue factor pathway inhibitor-2 inhibits endothelial cell migration and angiogenesis. AB - OBJECTIVE: Extracellular matrix (ECM) remodeling during angiogenesis is accomplished through plasmin-dependent pericellular proteolysis and through the action of matrix metalloproteinases (MMPs). Because tissue factor pathway inhibitor-2 (TFPI-2), a Kunitz-type protease inhibitor with prominent ECM localization, inhibits plasmin and MMPs activity, we investigated the role of TFPI-2 in endothelial cell (EC) migration and angiogenesis. METHODS AND RESULTS: Real-time polymerase chain reaction and immunostaining showed that the expression of TFPI-2 mRNA and protein was upregulated in migrating ECs. The effect of TFPI-2 on angiogenesis was studied in mouse models of Matrigel and polyvinylalcohol sponge implants by overexpressing TFPI-2 through infection with a replication deficient adenovirus (AdTFPI-2). Using (immuno)fluorescence and confocal microscopy we observed that TFPI-2 reduced neovascularization and promoted ECM deposition. Lateral cell migration and capillary tube formation in vitro also were impaired by TFPI-2, a process reversed by anti-TFPI-2 antibodies. Increased apoptosis occurred both in AdTFPI-2-treated ECs and in the mouse implants. Zymography and assays in the absence of plasminogen confirmed plasmin inhibition as a main mechanism through which TFPI-2 inhibits EC migration. CONCLUSIONS: Our data suggest that TFPI-2 may be an important regulator of aberrant angiogenesis associated with tumor growth/metastasis, cardiovascular diseases, chronic inflammation, or diabetes. PMID- 17138935 TI - Apolipoprotein E3- and nitric oxide-dependent modulation of endothelial cell inflammatory responses. AB - OBJECTIVE: Although apolipoprotein E3 (apoE3) is known to be atheroprotective, its mechanisms of protection in endothelial cells remain unclear. METHODS AND RESULTS: Cultured human aortic endothelial cells were stimulated with tumor necrosis factor (TNF)-alpha in the presence of human recombinant apoE3 solubilized in dimyristoyl phosphatidylcholine liposomes. Using flow cytometry and real-time polymerase chain reaction, a significant increase of inflammatory cell adhesion proteins (vascular cell adhesion molecule-1 and E-Selectin), and MCP-1, interleukin-8, and intercellular adhesion molecule-1 gene expression was observed within 5 hours of TNF-alpha exposure, which was markedly attenuated in cells coincubated with apoE3. Treatment with apoE4 resulted in increased inflammatory gene expression relative to either TNF treatment alone or TNF + apoE3 treatment. NO synthase inhibition experiments demonstrated NO to be an active participant in the actions of both TNF and apoE. To clarify the role of NO, dose-response experiments were performed with 0.03 to 300 micromol/L DEA NONOate. Using flow cytometry and real-time polymerase chain reaction, a modulatory role of NO in TNF-induced endothelial cell activation was observed. CONCLUSIONS: These data suggest a role of vascular wall apoE3 to balance the intracellular redox state in injured endothelial cells via NO-dependent pathways. PMID- 17138936 TI - In vivo plaque characterization using intravascular ultrasound-virtual histology in a porcine model of complex coronary lesions. AB - OBJECTIVE: To determine the accuracy of detection of different tissue types of intravascular ultrasound-virtual histology (IVUS-VH) in a porcine model of complex coronary lesions. METHODS AND RESULTS: Coronary lesions were induced by injecting liposomes containing human oxidized low-density lipoprotein into the adventitia of the arteries. IVUS-VH imaging was performed in vivo at 8.2+/-1.6 weeks after injection. A total of 60 vascular lesions were analyzed and compared with their correspondent IVUS-VH images. Correlation analysis was performed using linear regression models. Compared with histology, IVUS-VH correctly identified the presence of fibrous, fibro-fatty, and necrotic tissue in 58.33%, 38.33%, and 38.33% of lesions, respectively. The sensitivity of IVUS-VH for the detection of fibrous, fibro-fatty, and necrotic core tissue was 76.1%, 46%, and 41.1% respectively. A linear regression analysis performed for each individual plaque component did not show strong correlation that would allow significant prediction of individual values. CONCLUSIONS: In a porcine model of complex coronary lesions, IVUS-VH was not accurate in detecting the relative amount of specific plaque components within each individual corresponding histological specimen. PMID- 17138937 TI - Platelet activity, coagulation, and fibrinolysis during exercise in healthy males: effects of thrombin inhibition by argatroban and enoxaparin. AB - BACKGROUND: Relationships between exercise-induced activation of platelets, blood coagulation, and fibrinolysis, and the importance of thrombin for responses to exercise are not clear. METHODS AND RESULTS: Effects of thrombin inhibition on hemostatic parameters were examined in a double-blind crossover study comparing the direct thrombin inhibitor argatroban (350 microg/kg intravenous bolus followed by 25 microg/kg per minute of infusion), the indirect thrombin inhibitor enoxaparin (0.75 mg/kg, intravenous bolus), or placebo (saline) in 21 healthy males. Measurements were made at rest, before and during/after thrombin inhibitor treatment, and immediately after exhaustive exercise. At rest argatroban abolished, and enoxaparin attenuated platelet activation by thrombin, but not by adenosine diphosphate. Argatroban and, even more so, enoxaparin decreased thrombin generation (prothrombin F1+2) and the coagulation potential, and increased the fibrinolytic potential. Exercise increased circulating activated platelets (from 5.5+/-0.3 to 9.4+/-0.9x10(9)/L; P<0.001), circulating platelet platelet microaggregates, the platelet responsiveness to in vitro stimulation, leukocyte activation (leukocyte CD11b expression and plasma elastase), and platelet-leukocyte aggregation (P<0.01 for all). Exercise increased coagulation (F1+2; P<0.01) and fibrinolysis, but did not alter the balance between them; fibrin gel permeability increased (P<0.01), probably because of release of endogenous tissue plasminogen activator from the vessel wall. Neither argatroban nor enoxaparin counteracted exercise-induced platelet or leukocyte activation. Both thrombin inhibitors augmented exercise effects on fibrinolysis. CONCLUSIONS: Strenuous exercise enhances platelet and leukocyte activation independently of thrombin. Exercise augments both coagulation and fibrinolysis, but the balance between them appears to be maintained. At therapeutic dosages argatroban counteracted thrombin-induced platelet activation most efficiently, whereas enoxaparin had somewhat stronger anticoagulant and profibrinolytic effects. PMID- 17138938 TI - Mass-spectrometric identification of a novel angiotensin peptide in human plasma. AB - OBJECTIVE: Angiotensin peptides play a central role in cardiovascular physiology and pathology. Among these peptides, angiotensin II (Ang II) has been investigated most intensively. However, further angiotensin peptides such as Ang 1-7, Ang III, and Ang IV also contribute to vascular regulation, and may elicit additional, different, or even opposite effects to Ang II. Here, we describe a novel Ang II-related, strong vasoconstrictive substance in plasma from healthy humans and end-stage renal failure patients. METHODS AND RESULTS: Chromatographic purification and structural analysis by matrix-assisted laser desorption/ionisation time-of-flight/time-of-flight (MALDI-TOF/TOF) revealed an angiotensin octapeptide with the sequence Ala-Arg-Val-Tyr-Ile-His-Pro-Phe, which differs from Ang II in Ala1 instead of Asp1. Des[Asp1]-[Ala1]-Ang II, in the following named Angiotensin A (Ang A), is most likely generated enzymatically. In the presence of mononuclear leukocytes, Ang II is converted to Ang A by decarboxylation of Asp1. Ang A has the same affinity to the AT1 receptor as Ang II, but a higher affinity to the AT2 receptor. In the isolated perfused rat kidney, Ang A revealed a smaller vasoconstrictive effect than Ang II, which was not modified in the presence of the AT2 receptor antagonist PD 123319, suggesting a lower intrinsic activity at the AT1 receptor. Ang II and Ang A concentrations in plasma of healthy subjects and end-stage renal failure patients were determined by matrix-assisted laser desorption/ionisation mass-analysis, because conventional enzyme immunoassay for Ang II quantification did not distinguish between Ang II and Ang A. In healthy subjects, Ang A concentrations were less than 20% of the Ang II concentrations, but the ratio Ang A/Ang II was higher in end-stage renal failure patients. CONCLUSIONS: Ang A is a novel human strong vasoconstrictive angiotensin-derived peptide, most likely generated by enzymatic transformation through mononuclear leukocyte-derived aspartate decarboxylase. Plasma Ang A concentration is increased in end-stage renal failure. Because of its stronger agonism at the AT2 receptor, Ang A may modulate the harmful effects of Ang II. PMID- 17138939 TI - Ccr5 but not Ccr1 deficiency reduces development of diet-induced atherosclerosis in mice. AB - OBJECTIVE: Chemokines and their receptors are crucially involved in the development of atherosclerotic lesions by directing monocyte and T cell recruitment. The CC-chemokine receptors 1 (CCR1) and 5 (CCR5) expressed on these cells bind chemokines implicated in atherosclerosis, namely CCL5/RANTES. Although general blockade of CCL5 receptors reduces atherosclerosis, specific roles of CCR1 and CCR5 have not been unequivocally determined. METHODS AND RESULTS: We provide two independent lines of investigation to dissect the effects of Ccr1 and Ccr5 deletion in apolipoprotein E-deficient (ApoE-/-) mice in a collaboration between Aachen/Germany and Geneva/Switzerland. Different strains of ApoE-/- Ccr5 /- mice, ApoE-/- Ccr1-/- mice or respective littermates, were fed a high-fat diet for 10 to 12 weeks. Plaque areas were quantified in the aortic roots and thoracoabdominal aortas. Concordantly, both laboratories found that lesion formation was reduced in ApoE-/- Ccr5-/- mice. Plaque quality and immune cells were assessed by immunohistochemistry or mRNA analysis. Whereas lesional macrophage content, aortic CD4, and Th1-related Tim3 expression were reduced, smooth muscle cell (SMC) content and expression of interleukin-10 in plaques, lesional SMCs, and splenocytes were elevated. Protection against lesion formation by Ccr5 deficiency was sustained over 22 weeks of high-fat diet or over 26 weeks of chow diet. Conversely, plaque area, T cell, and interferon-gamma content were increased in ApoE-/- Ccr1-/- mice. CONCLUSIONS: Genetic deletion of Ccr5 but not Ccr1 in ApoE-/- mice protects from diet-induced atherosclerosis, associated with a more stable plaque phenotype, reduced mononuclear cell infiltration, Th1-type immune responses, and increased interleukin-10 expression. This corroborates CCR5 as a promising therapeutic target. PMID- 17138940 TI - Cerebrovascular nitrosative stress mediates neurovascular and endothelial dysfunction induced by angiotensin II. AB - OBJECTIVE: Angiotensin II (AngII) disrupts the regulation of the cerebral circulation through superoxide, a reactive oxygen species (ROS) generated by a nox2-containing NADPH oxidase. We tested the hypothesis that AngII-derived superoxide reacts with nitric oxide (NO) to form peroxynitrite, which, in turn, contributes to the vascular dysfunction. METHODS AND RESULTS: Cerebral blood flow (CBF) was monitored by laser Doppler flowmetry in the neocortex of anesthetized mice equipped with a cranial window. AngII (0.25+/-0.02 microg/kg/min; intravenous for 30 to 45 minutes) attenuated the cerebral blood flow (CBF) increase produced by topical application of the endothelium-dependent vasodilator acetylcholine (-43+/-1%) and by whisker stimulation (-47+/-1%). AngII also increased the nitration marker 3-nitrotyrosine (3-NT) in cerebral blood vessels, an effect dependent on NO and nox2-derived ROS. Both the cerebrovascular effects of AngII and the nitration were attenuated by pharmacological inhibition or genetic inactivation of NO synthase. The nitration inhibitor uric acid or the peroxynitrite decomposition catalyst FeTPPS abolished AngII-induced cerebrovascular nitration and prevented the cerebrovascular effects of AngII. CONCLUSIONS: These findings provide evidence that peroxynitrite, formed from NO and nox2-derived superoxide, contributes to the deleterious cerebrovascular effects of AngII. Inhibitors of peroxynitrite action may be valuable tools to counteract the deleterious cerebrovascular effects of AngII-induced hypertension. PMID- 17138941 TI - Implications of early structural-functional changes in the endothelium for vascular disease. AB - By location, between the blood and tissues and the multiple functions, the endothelial cells (ECs) play a major role in securing body homeostasis. The ECs sense all variations occurring in the plasma and interstitial fluid, and respond (function of intensity), initially by modulation of their constitutive functions, then by dysfunction, expressed by temporarily altered functions and a phenotypic shift, and ultimately by injury/death. In dyslipidemia/hyperglycemia, the initial response of EC is the modulation of 2 constitutive functions: permeability and biosynthesis. Increased transcytosis of plasma beta-lipoproteins leads to their accumulation within the hyperplasic basal lamina, interaction with matrix proteins, and conversion to modified and reassembled lipoproteins (MRL). This generates a multipart inflammatory process and EC dysfunction characterized by expression of new cell adhesion molecules and MCP-1 that trigger T-lymphocytes and monocyte recruitment, diapedesis, and homing within the subendothelium where activated macrophages become foam cells. The latter, together with the subendothelial accrual of MRL, growth factors, cytokines, and chemokines, and accretion of smooth muscle cells of various sources lead to atheroma formation; in advanced disease, the EC overlaying atheroma take up lipids, become EC-derived foam cells, and the cytotoxic ambient ultimately conducts to EC apoptosis. Understanding the mechanisms of EC dysfunction is a prerequisite for EC-targeted therapy to reduce the incidence of cardiovascular diseases. PMID- 17138942 TI - Antiangiogenic activity of the MDM2 antagonist nutlin-3. AB - Nutlin-3, a nongenotoxic activator of the p53 pathway, dose-dependently (range 0.1 to 10 micromol/L) inhibited the formation of capillaries in an in vivo matrigel assay, as well as the formation of capillary-like structures in an in vitro coculture system composed of endothelial cells surrounded by fibroblasts. In contrast to the chemotherapeutic agent doxorubicin, nutlin-3 showed no induction of apoptosis in vitro either in the cocultures or in isolated vascular endothelial cells, even when used at the highest concentration (10 micromol/L). However, treatment with pharmacological inhibitors of the nuclear factor kappaB and phosphatidylinositol 3-kinase/Akt pathways sensitized endothelial cells to nutlin-3-induced apoptosis. Although nutlin-3 and doxorubicin induced a comparable p53 accumulation in endothelial cells, nutlin-3 was significantly more efficient than doxorubicin in upregulating the p53 target genes CDKN1A/p21, MDM2, and GDF-15, as well as in inhibiting cell cycle progression. However, the predominant in vitro effect of nutlin-3 was its strong antimigratory activity observed at concentrations significantly lower (0.1 micromol/L) than those required to inhibit endothelial cell cycle progression. Taken together, our data suggest that the antiangiogenic activity of nutlin-3 observed in vivo was mainly attributable to inhibition of endothelial cell migration, to some extent attributable to cell cycle arrest, and to a lesser extent attributable to induction of apoptosis. PMID- 17138943 TI - Nitroxyl improves cellular heart function by directly enhancing cardiac sarcoplasmic reticulum Ca2+ cycling. AB - Heart failure remains a leading cause of morbidity and mortality worldwide. Although depressed pump function is common, development of effective therapies to stimulate contraction has proven difficult. This is thought to be attributable to their frequent reliance on cAMP stimulation to increase activator Ca(2+). A potential alternative is nitroxyl (HNO), the 1-electron reduction product of nitric oxide (NO) that improves contraction and relaxation in normal and failing hearts in vivo. The mechanism for myocyte effects remains unknown. Here, we show that this activity results from a direct interaction of HNO with the sarcoplasmic reticulum Ca(2+) pump and the ryanodine receptor 2, leading to increased Ca(2+) uptake and release from the sarcoplasmic reticulum. HNO increases the open probability of isolated ryanodine-sensitive Ca(2+)-release channels and accelerates Ca(2+) reuptake into isolated sarcoplasmic reticulum by stimulating ATP-dependent Ca(2+) transport. Contraction improves with no net rise in diastolic calcium. These changes are not induced by NO, are fully reversible by addition of reducing agents (redox sensitive), and independent of both cAMP/protein kinase A and cGMP/protein kinase G signaling. Rather, the data support HNO/thiolate interactions that enhance the activity of intracellular Ca(2+) cycling proteins. These findings suggest HNO donors are attractive candidates for the pharmacological treatment of heart failure. PMID- 17138944 TI - Cardiomyocyte-specific overexpression of nitric oxide synthase 3 prevents myocardial dysfunction in murine models of septic shock. AB - Myocardial dysfunction contributes to the high mortality of patients with endotoxemia. Although nitric oxide (NO) has been implicated in the pathogenesis of septic cardiovascular dysfunction, the role of myocardial NO synthase 3 (NOS3) remains incompletely defined. Here we show that mice with cardiomyocyte-specific NOS3 overexpression (NOS3TG) are protected from myocardial dysfunction and death associated with endotoxemia. Endotoxin induced more marked impairment of Ca(2+) transients and cellular contraction in wild-type than in NOS3TG cardiomyocytes, in part, because of greater total sarcoplasmic reticulum Ca(2+) load and myofilament sensitivity to Ca(2+) in the latter during endotoxemia. Endotoxin increased reactive oxygen species production in wild-type but not NOS3TG hearts, in part, because of increased xanthine oxidase activity. Inhibition of NOS by N(G)-nitro-l-arginine-methyl ester restored the ability of endotoxin to increase reactive oxygen species production and xanthine oxidase activity in NOS3TG hearts to the levels measured in endotoxin-challenged wild-type hearts. Allopurinol, a xanthine oxidase inhibitor, attenuated endotoxin-induced reactive oxygen species accumulation and myocardial dysfunction in wild-type mice. The protective effects of cardiomyocyte NOS3 on myocardial function and survival were further confirmed in a murine model of polymicrobial sepsis. These results suggest that increased myocardial NO levels attenuate endotoxin-induced reactive oxygen species production and increase total sarcoplasmic reticulum Ca(2+) load and myofilament sensitivity to Ca(2+), thereby reducing myocardial dysfunction and mortality in murine models of septic shock. PMID- 17138945 TI - The socioeconomic stroke puzzle. PMID- 17138946 TI - Accuracy of the Siriraj and Guy's Hospital Stroke Scores in urban South Africans. AB - BACKGROUND AND PURPOSE: The burden of stroke in sub-Saharan Africa is already high and likely to increase, but few patients with stroke have access to brain imaging. Distinguishing pathologic stroke types is relevant both for clinical management and epidemiologic studies. We assessed the accuracy of two stroke scores in distinguishing stroke types in a population known to have a high prevalence of intracranial hemorrhage but low prevalence of atherosclerosis and compared them with the clinicians' assessment of stroke type with computed tomography brain scanning as the "gold standard." METHODS: We assessed the stroke scores and the clinicians' blind assessment of pathologic stroke type in consecutive black patients with stroke included in the Johannesburg Hospital Stroke Register over 23 months. We calculated the accuracy of the scores and clinicians compared with computed tomography brain scan (sensitivity, specificity, positive predictive value, likelihood ratio, kappa statistic). RESULTS: Two hundred twenty-two patients were scanned and assessed within 15 days. Sixty-two (28%) had cerebral hemorrhage and nine (4%) subarachnoid hemorrhage. Neither the Siriraj (sub-Saharan Africa) nor Guy's Hospital score was accurate or offered much advantage over clinician assessment (sensitivity 0.60 and 0.34, specificity 0.88 and 0.95 for intracranial hemorrhage in the Siriraj Stroke Score and Guy's Hospital Stroke Score, respectively; sensitivity 0.70 and 0.71, specificity 0.84 and 0.74, respectively, for ischemic stroke). Although the scores were more accurate when we used new cutoff points, they then failed to diagnose over 80% of stroke types. CONCLUSIONS: The Siriraj Stroke Score and Guy's Hospital Stroke Score are not sufficiently accurate for use in either epidemiologic studies or to guide clinical management in sub-Saharan Africa at present. PMID- 17138947 TI - Relation of adult height with stroke mortality in Japan: NIPPON DATA80. AB - BACKGROUND AND PURPOSE: The age-adjusted stroke mortality rate in Japan was the highest in the world from 1950 to the 1970s, but it started to dramatically decrease after 1965. In addition to improved management of high blood pressure, the increase in average height might also contribute to this reduction. The present study investigates whether height is an independent risk for stroke mortality in Japan. METHODS: Among participants of the National Survey on Cardiovascular Diseases in 1980 who were randomly selected from the Japanese population, we followed up 3969 and 4955 Japanese men and women without prior cardiovascular disease for a maximum of 19 years and observed 158 and 132 stroke deaths. RESULTS: Height was inversely correlated with age and with crude stroke mortality. The relationship was attenuated in men when we adjusted for age or other possible confounders (multivariate adjusted relative hazards of a 5-cm increase of height for stroke mortality: 0.92, 95% CI: 0.79 to 1.08). For women, the inverse relationship (relative hazard: 0.77: 95% CI: 0.64 to 0.91) remained after multivariate adjustment. These relationships persisted when we stratified participants by age. CONCLUSIONS: Height is inversely related to stroke mortality and the relationship is statistically significant among Japanese women. PMID- 17138948 TI - The socioeconomic gradient in the incidence of stroke: a prospective study in middle-aged women in Sweden. AB - BACKGROUND AND PURPOSE: A socioeconomic gradient in stroke has been demonstrated in a variety of settings, but mostly in men. Our purpose was to establish whether a socioeconomic gradient in stroke existed in a group of Swedish women and whether this gradient could be explained by established stroke risk factors or psychosocial factors. METHODS: The Women's Lifestyle and Health Cohort Study includes 49 259 women from Sweden aged 30 to 50 years at baseline (1991 to 1992). The women completed an extensive questionnaire and were traced through linkages to national registries until the end of 2002. Among the 47 942 women included in these analyses, there were 200 cases of incident stroke during follow up (121 ischemic stroke, 47 hemorrhagic stroke, and 32 of unknown origin). Statistical analysis was through the Cox proportional hazards model. RESULTS: The risk of stroke was significantly inversely related to years of education completed, our proxy for socioeconomic status (hazard ratio comparing lowest with highest education group=2.1, 95% CI: 1.4 to 2.9, P for trend <0.001). This association was reduced after adjustment for established risk factors, although remaining significant (1.5, 1.0 to 2.2, P for trend=0.04). The gradient was more pronounced for ischemic stroke (2.9, 1.8 to 4.7, P for trend <0.001) than for hemorrhagic stroke (1.4, 0.7 to 2.9, P for trend=0.35). Job strain and social support were unrelated to risk of stroke. Self-rated health was strongly related to risk of stroke mediated by established risk factors. Psychosocial factors did not contribute toward the socioeconomic gradient in stroke. CONCLUSIONS: There was a strong gradient in risk of stroke by years of education, especially for ischemic stroke. Most of the social gradient was explained by established risk factors, particularly smoking and alcohol, but not by psychosocial factors. PMID- 17138949 TI - Clinical deterioration after intravenous recombinant tissue plasminogen activator treatment: a multicenter transcranial Doppler study. AB - BACKGROUND AND PURPOSE: Patients may experience clinical deterioration (CD) after treatment with intravenous recombinant tissue plasminogen activator (rt-PA). We evaluated the ability of flow findings on transcranial Doppler to predict CD and outcomes on modified Rankin Scale. METHODS: Patients with acute stroke received intravenous rt-PA within 3 hours of symptom onset at four academic centers. CD was defined as an increase in the National Institutes of Health Stroke Scale (NIHSS) score by 4 points or more within 24 hours. Poor long-term outcome was defined by modified Rankin Scale > or =2 at 3 months. Transcranial Doppler findings were interpreted using the Thrombolysis in Brain Ischemia flow grading system as persistent arterial occlusion, reocclusion, or complete recanalization. Multiple regression analysis was used to identify transcranial Doppler flow as a predictor for CD after controlling for age, sex, baseline NIHSS, hypertension, and glucose. RESULTS: A total of 374 patients received intravenous rt-PA at 142+/ 60 minutes (median pretreatment NIHSS score 16 points). At the end of intravenous rt-PA infusion, transcranial Doppler showed persistent arterial occlusion in 219 patients (59%), arterial reocclusion in 54 patients (14%), and complete recanalization in 101 patients (27%). CD occurred in 44 patients: 36 had persistent arterial occlusion or reocclusion (82%), 13 symptomatic intracerebral hemorrhage (29%), and both persistent occlusion/reocclusion and symptomatic intracerebral hemorrhage in 10 patients (23%). After adjustment, patient risk for CD with persistent occlusion was OR 1.7 (95% CI: 0.7 to 4) and with arterial reocclusion 4.9 (95% CI: 1.7 to 13) (P=0.002). Patient risk for poor long-term outcomes with persistent occlusion, partial recanalization, or reocclusion was OR 5.2 (95% CI: 2.7 to 9, P=0.001). CONCLUSIONS: Inability to achieve or sustain vessel patency at the end of rt-PA infusion correlates with the likelihood of clinical deterioration and poor long-term outcome. Early arterial reocclusion on transcranial Doppler is highly predictive of CD and poor outcome. PMID- 17138950 TI - Increased body iron stores are associated with poor outcome after thrombolytic treatment in acute stroke. AB - BACKGROUND AND PURPOSE: Iron overload has been associated with greater oxidative stress and brain injury in experimental cerebral ischemia and reperfusion. This study investigates whether high serum ferritin levels, as an index of increased cellular iron stores, are associated with poor outcome, hemorrhagic transformation, and brain edema after treatment with tissue plasminogen activator in patients with acute ischemic stroke. METHODS: A total of 134 consecutive patients treated with intravenous tissue plasminogen activator were prospectively studied in four centers. Serum ferritin levels were determined at baseline, 24 and 72 hours after treatment. Cranial computed tomography was performed on admission and at 24 to 36 hours after tissue plasminogen activator infusion. Stroke severity and outcome were evaluated by using the National Institute of Health Stroke Scale and the modified Rankin Scale. RESULTS: Computed tomography showed hemorrhagic transformation in 27 patients (hemorrhagic infarction in 15 and parenchymal hematoma in 12; symptomatic in four) and brain swelling with midline shift in 15. Poor outcome (modified Rankin Scale >2) at 90 days was observed in 54.5% of patients. Ferritin levels at baseline were higher in patients with poor outcome at 90 days (median [quartiles], 165 [98,307] versus 17 [12,37] ng/mL; P<0.001) and in those who developed parenchymal hematoma (P=0.006), symptomatic hemorrhagic transformation (P=0.008), and severe brain edema (P<0.001). Serum ferritin levels higher than 79 ng/mL before tissue plasminogen activator treatment were independently associated with poor outcome (OR, 117 [95% CI, 25 to 557]). CONCLUSIONS: Increased body iron stores are associated with poor outcome, symptomatic hemorrhagic transformation, and severe edema in patients treated with tissue plasminogen activator after ischemic stroke. These findings suggest that iron overload may offset the beneficial effect of thrombolytic therapies. PMID- 17138951 TI - Smoking and the platelet fibrinogen receptor glycoprotein IIb/IIIA PlA1/A2 polymorphism interact in the risk of lacunar stroke and midterm survival. AB - BACKGROUND AND PURPOSE: Smoking, increased fibrinogen levels, and platelet activation are related to the risk of ischemic stroke. The platelet fibrinogen receptor glycoprotein (Gp) IIb/IIIa Pl(A1/A2) polymorphism affects the binding of platelets to fibrinogen and is suggested to interact with smoking. METHODS: We explored the association of smoking and the Pl(A1/A2) polymorphism with ischemic stroke and survival in the Stroke Aging Memory cohort, comprising 486 consecutive patients (55 to 85 years old) who were analyzed 3 months after an ischemic stroke and followed up for 15 months. Stroke subtype determined by magnetic resonance imaging and GpIIb/IIIa Pl(A1/A2) genotype data were available for 272 patients. RESULTS: In multivariate analysis, smoking was the only factor related to the risk of lacunar infarcts (odds ratio [OR]=1.87, 95% CI=1.05 to 3.31; P=0.033), and it was also a predictor of death (n=24, 8.8%) at 15 months (OR=5.13, 95% CI=1.61 to 16.36; P=0.006), along with age (OR=1.10, 95% CI=1.01 to 1.19; P=0.008). The GpIIb/IIIa Pl(A1/A2) polymorphism alone showed no association with stroke subtype or survival. However, there was a smoking-by-genotype association with the risk of lacunar infarcts (OR=2.10, 95% CI=0.90 to 4.89; P=0.087) and with survival (OR=2.78, 95% CI=0.89 to 8.61; P=0.077). Among younger (55 to 69 years) stroke patients, smokers carrying the Pl(A2) allele were at a higher (OR=5.81, 95% CI=1.26 to 26.80; P=0.024) risk of lacunar infarcts than noncarrier smokers (OR=3.12, 95% CI=1.06 to 9.24; P=0.039). The effect of Pl(A2) and smoking combined on survival was also stronger (OR=8.86, 95% CI=1.68 to 46.55; P=0.010) than the effect of smoking alone (OR=5.06, 95% CI=1.20 to 21.35; P=0.027). CONCLUSIONS: Our results indicate that prothrombotic genetic factors may interact with smoking by modifying the stroke phenotype and affecting midterm survival. PMID- 17138952 TI - Depressive symptoms and risk of stroke: the Framingham Study. AB - BACKGROUND AND PURPOSE: Emerging evidence raises the possibility of an association between depression and stroke risk. This study sought to examine whether depressive symptoms are associated with an increased risk of cerebrovascular events in a community-based sample. METHODS: A prospective study was conducted on 4120 Framingham Heart Study participants aged 29 to 100 years with up to 8 years of follow-up. The Center for Epidemiologic Studies Depression Scale was used to measure depressive symptoms. Incident stroke and transient ischemic attack (TIA) events were assessed by uniform diagnostic criteria. The association between depressive symptoms and risk of stroke/TIA was analyzed with Cox proportional-hazards models, after adjusting for traditional stroke risk factors. RESULTS: In participants <65 years, the risk of developing stroke/TIA was 4.21 times greater (P= <0.001) in those with symptoms of depression. After adjusting for components of the Framingham Stroke Risk Profile (hazard ratio=3.43, 95% CI=1.60 to 7.36, P=0.002) and education (hazard ratio=4.89, 95% CI=2.19 to 10.95), similar results were obtained. In subjects aged 65 and older, depressive symptoms were not associated with an increased risk of stroke/TIA. Taking antidepressant medications did not alter the risk associated with depressive symptoms. CONCLUSIONS: In this community-based study, depressive symptoms were an independent risk factor for incident stroke/TIA in individuals <65 years. These data suggest that identification of depressive symptoms at younger ages may have an impact on the primary prevention of stroke. PMID- 17138953 TI - Are mood disorders a stroke risk factor? PMID- 17138954 TI - Targeting the networks that underpin contiguous immunity in asthma and chronic obstructive pulmonary disease. AB - Recent advances in the field of innate immunity have driven an important reappraisal of the role of these processes in airway disease. Various strands of evidence indicate that resident cells, such as macrophages and epithelial cells, have central importance in the initiation of inflammation. Macrophage activation has the potential to regulate not just typical aspects of innate immunity but also, via a variety of intricate cell-cell networks, adaptive responses and responses characterized by Th2-type cytokine production. In turn, such adaptive immune processes modify the phenotype and function of the innate immune system. Cooperative responses between monocytic cells and tissue cells are likely to be crucial to the generation of effective inflammatory responses, and a realization of the importance of these networks is providing a new way of identifying antiinflammatory therapies. Importantly, the repeated cycles of allergic and nonallergic inflammation that comprise chronic human airway disease are not necessarily well described by current terminology, and we propose and describe a concept of contiguous immunity, in which continual bidirectional cross-talk between innate and adaptive immunity describes disease processes more accurately. PMID- 17138955 TI - Impact of intensive insulin therapy on neuromuscular complications and ventilator dependency in the medical intensive care unit. AB - RATIONALE: Critical illness polyneuropathy/myopathy causes limb and respiratory muscle weakness, prolongs mechanical ventilation, and extends hospitalization of intensive care patients. Besides controlling risk factors, no specific prevention or treatment exists. Recently, intensive insulin therapy prevented critical illness polyneuropathy in a surgical intensive care unit. OBJECTIVES: To investigate the impact of intensive insulin therapy on polyneuropathy/myopathy and treatment with prolonged mechanical ventilation in medical patients in the intensive care unit for at least 7 days. METHODS: This was a prospectively planned subanalysis of a randomized controlled trial evaluating the effect of intensive insulin versus conventional therapy on morbidity and mortality in critically ill medical patients. All patients who were still in intensive care on Day 7 were screened weekly by electroneuromyography. The effect of intensive insulin therapy on critical illness polyneuropathy/myopathy and the relationship with duration of mechanical ventilation were assessed. MEASUREMENTS AND MAIN RESULTS: Independent of risk factors, intensive insulin therapy reduced incidence of critical illness polyneuropathy/myopathy (107/212 [50.5%] to 81/208 [38.9%], p = 0.02). Treatment with prolonged (> or = 14 d) mechanical ventilation was reduced from 99 of 212 (46.7%) to 72 of 208 (34.6%) (p = 0.01). This was statistically only partially explained by prevention of critical illness polyneuropathy/myopathy. CONCLUSION: In a subset of medical patients in the intensive care unit for at least 7 days, enrolled in a randomized controlled trial of intensive insulin therapy, those assigned to intensive insulin therapy had a reduced incidence of critical illness polyneuropathy/myopathy and were treated with prolonged mechanical ventilation less frequently. PMID- 17138956 TI - Increased granzyme A expression in type II pneumocytes of patients with severe chronic obstructive pulmonary disease. AB - RATIONALE: Chronic obstructive pulmonary disease (COPD) is associated with increased numbers of CD8(+) cytotoxic T lymphocytes (CTLs) in the lung, but the functional activity of CTLs remains unknown. Granzyme A (GrA) and B (GrB) are serine proteases considered to be important effector molecules of CTLs and natural killer cells. OBJECTIVE: To investigate protein and mRNA expression of GrA and GrB in peripheral lung tissue from patients with COPD and control subjects with normal lung function. METHODS: Paraffin-embedded sections of surgical lung specimens from 22 patients with COPD (FEV(1), 22% predicted; GOLD stage 4) and 15 control subjects (FEV(1), 108% predicted) were immunostained for GrA and GrB, and semiquantified on a 3-point scale. Messenger RNA expression in total lung, specific cell types enriched for by laser capture microdissection, and freshly isolated primary cells were determined by reverse transcriptase polymerase chain reaction. MEASUREMENTS AND MAIN RESULTS: GrA and GrB immunoreactivity was observed in CD8(+) CTLs and CD57(+) natural killer cells, but also in type II pneumocytes and alveolar macrophages in both groups. Bronchiolar epithelium stained positive for GrA, but negative for GrB. These observations were confirmed by reverse transcriptase-polymerase chain reaction on total lung, laser capture microdissection-enriched specific cell types and freshly isolated primary type II pneumocytes. The scores of GrA-expressing type II pneumocytes were significantly higher in patients with COPD versus control subjects. CONCLUSIONS: GrA and GrB mRNA and protein are detectable in human lung tissue. GrA expression is increased in type II pneumocytes of patients with very severe COPD. These results indicate that GrA may be important in the development of COPD. PMID- 17138957 TI - Down-regulation of CXCR2 on neutrophils in severe sepsis is mediated by inducible nitric oxide synthase-derived nitric oxide. AB - RATIONALE: The failure of neutrophils to migrate to an infection focus during severe sepsis is an important determinant of the inability of a host to deal with an infectious insult. Our laboratory has shown that inducible nitric oxide synthase (iNOS) induction and NO production contribute to the failure of neutrophils to migrate in the context of sepsis. OBJECTIVES AND METHODS: We investigated whether CXCR2 expression contributed to the failure of neutrophils to migrate during severe sepsis and the role of NO in modulating CXCR2 expression on neutrophils in mice subjected to nonsevere (NS) or severe (S) cecal ligation and puncture (CLP). RESULTS: Neutrophil migration to the infection focus was deficient in S-CLP mice, a phenomenon prevented by pharmacologic (aminoguanidine, l-canavanine) or genetic (iNOS gene deletion) inhibition of iNOS. The expression of CXCR2 on neutrophils from S-CLP mice was significantly reduced when compared with neutrophils from NS-CLP or sham-operated mice. CXCR2 expression was reestablished by pharmacologic and genetic inhibition of iNOS. Immunofluorescence and confocal analysis revealed that iNOS blockade reduced neutrophil CXCR2 internalization. Adhesion and emigration of neutrophils in macrophage inflammatory protein-2-stimulated mesentery microcirculation were reduced in S CLP mice, compared with NS-CLP mice, and reestablished by pretreatment with aminoguanidine or l-canavanine. The NO donor S-nitroso-N-acetyl-d,l-penicillamine inhibited CXCL8-induced human neutrophil chemotaxis and CXCR2 expression on human and murine neutrophils. CONCLUSION: These results highlight evidences that the failure of neutrophils to migrate to an infection focus during severe sepsis is associated with excessive NO production and NO-dependent regulation of the expression of CXCR2 on the neutrophil surface. PMID- 17138958 TI - Enhanced green fluorescent protein is a nearly ideal long-term expression tracer for hematopoietic stem cells, whereas DsRed-express fluorescent protein is not. AB - Validated gene transfer and expression tracers are essential for elucidating functions of mammalian genes. Here, we have determined the suitability and unintended side effects of enhanced green fluorescent protein (EGFP) and DsRed Express fluorescent protein as expression tracers in long-term hematopoietic stem cells (HSCs). Retrovirally transduced mouse bone marrow cells expressing either EGFP or DsRed-Express in single or mixed dual-color cell populations were clearly discerned by flow cytometry and fluorescence microscopy. The results from in vivo competitive repopulation assays demonstrated that EGFP-expressing HSCs were maintained nearly throughout the lifespan of the transplanted mice and retained long-term multilineage repopulating potential. All mice assessed at 15 months post-transplantation were EGFP positive, and, on average, 24% total peripheral white blood cells expressed EGFP. Most EGFP-expressing recipient mice lived at least 22 months. In contrast, Discosoma sp. red fluorescent protein (DsRed) expressing donor cells dramatically declined in transplant-recipient mice over time, particularly in the competitive setting, in which mixed EGFP- and DsRed expressing cells were cotransplanted. Moreover, under in vitro culture condition favoring preservation of HSCs, purified EGFP-expressing cells grew robustly, whereas DsRed-expressing cells did not. Therefore, EGFP has no detectable deteriorative effects on HSCs, and is nearly an ideal long-term expression tracer for hematopoietic cells; however, DsRed-Express fluorescent protein is not suitable for these cells. PMID- 17138959 TI - Concise review: cancer/testis antigens, stem cells, and cancer. AB - In the multistep process of cancer development, the concept that cancer stem cells are derived from normal stem cells that have gradually accumulated various genetic and epigenetic defects is gaining strong evidence. A number of investigations have identified molecular markers that, under normal conditions, are responsible for stem cell homeostasis but are also expressed in tumor "stem cell-like" subpopulations. In this regard, it was recently reported that a group of tumor-specific antigens known as cancer/testis antigens (CTAs) are expressed in human MSCs. It has long been stated that in normal tissue these antigens are exclusively expressed in germ cell precursors; however, based on these results, we suggest that CTAs are expressed at earlier stages during embryogenesis. The tumor-restricted expression of CTAs has led to several immunotherapeutic trials targeting some of these proteins. The clinical implications that these trials may have on the normal stem cell pools, as well as the immunologic properties of these cells, is to date poorly studied and should be considered. PMID- 17138960 TI - Synovial stem cells are regionally specified according to local microenvironments after implantation for cartilage regeneration. AB - We previously demonstrated that synovium-derived MSCs had greater in vitro chondrogenic ability than other mesenchymal tissues, suggesting a superior cell source for cartilage regeneration. Here, we transplanted undifferentiated synovium-derived MSCs into a full-thickness articular cartilage defect of adult rabbits and defined the cellular events to elucidate the mechanisms that govern multilineage differentiation of MSCs. Full-thickness osteochondral defects were created in the knee; the defects were filled with 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate-labeled MSCs and covered with periosteum. After 4 weeks, although the cell density decreased, transplanted MSCs produced a great amount of cartilage matrix extensively. The periosteum became thinner, and chondroprogenitors in the periosteum produced a small amount of cartilage matrix. In the deeper zone, transplanted MSCs progressed to the hypertrophic chondrocyte like cells. In the deep zone, some transplanted cells differentiated into bone cells and were replaced with host cells thereafter. In the next phase, the border between bone and cartilage moved upwards. In addition, integrations between native cartilage and regenerated tissue were improved. Chondrocyte-like cells derived from the transplanted MSCs still remained at least after 24 weeks. Histological scores of the MSC group improved continuously and were always better than those of two other control groups. Immunohistological analyses and transmission electron microscopy confirmed that the MSCs produced abundant cartilage matrix. We demonstrated that transplanted synovium-derived MSCs were altered over a time course according to the microenvironments. Our results will advance MSC-based therapeutic strategies for cartilage injury and provide the clues for the mechanisms that govern multilineage differentiation of MSCs. PMID- 17138961 TI - Constitutive gene expression predisposes morphogen-mediated cell fate responses of NT2/D1 and 27X-1 human embryonal carcinoma cells. AB - Human embryonal carcinoma (EC) cell lines exhibit considerable heterogeneity in their levels of pluripotency. Thus, NT2/D1 cells differentiate into neural lineages upon exposure to all-trans retinoic acid (ATRA) and non-neural epithelial lineages upon exposure to bone morphogenetic protein-2 (BMP-2). In contrast, 27X-1 cells differentiate into extra-embryonic endodermal (ExE) cells upon treatment with either morphogen. To understand the molecular basis for the differential responses of the two cell lines, we performed gene expression profiling at the undifferentiated EC cell line state to identify constitutive differences in gene expression. NT2/D1 cells preferentially expressed transcripts associated with neurectodermal development, whereas 27X-1 cells expressed high levels of transcripts associated with mesendodermal characteristics. We then determined temporal expression profiles of 27X-1 cells during ExE differentiation upon treatment with ATRA and BMP-2 and compared the data with changes in gene expression observed during BMP-2- and ATRA-induced differentiation of NT2/D1 cells. ATRA and BMP-2 induced distinct sets of transcription factors and phenotypic markers in the two EC cell lines, underlying distinct lineage choices. Although 27X-1 differentiation yielded comprehensive gene expression profiles of parietal endodermal lineages, we were able to use the combined analysis of 27X-1 data with data derived from yolk sac tumors for the identification of transcripts associated with visceral endoderm formation. Our results demonstrate constitutive differences in the levels of pluripotency between NT2/D1 and 27X-1 cells that correlate with lineage potential. This study also demonstrates that EC cells can serve as robust models to investigate early lineage choices during both embryonic and extra-embryonic human development. PMID- 17138962 TI - Bone morphogenetic protein-4 enhances cardiomyocyte differentiation of cynomolgus monkey ESCs in knockout serum replacement medium. AB - Despite extensive research in the differentiation of rodent ESCs into cardiomyocytes, there have been few studies of this process in primates. In this study, we examined the role of bone morphogenic protein-4 (BMP-4) to induce cardiomyocyte differentiation of cynomolgus monkey ESCs. To study the role of BMP 4, EBs were formed and cultured in Knockout Serum Replacement (KSR) medium containing BMP-4 for 8 days and subsequently seeded in gelatin-coated dishes for 20 days. It was found that ESCs differentiated into cardiomyocytes upon stimulation with BMP-4 in KSR medium, which resulted in a large fraction of beating EBs ( approximately 16%) and the upregulation of cardiac-specific proteins in a dose and time-dependent manner. In contrast, the addition of BMP-4 in FBS-containing medium resulted in a lower fraction of beating EBs ( approximately 6%). BMP-4 acted principally between mesendodermal and mesoderm progenitors and subsequently enhanced their expression. Ultrastructural observation revealed that beating EBs contained mature cardiomyocytes with sarcomeric structures. In addition, immunostaining, reverse transcription polymerase chain reaction, and Western blotting for cardiac markers confirmed the increased differentiation of cardiomyocytes in these cultures. Moreover, electrophysiological studies demonstrated that the differentiated cardiomyocytes were electrically activated. These findings may be useful in developing effective culture conditions to differentiate cynomolgus monkey ESCs into cardiomyocytes for studying developmental biology and for regenerative medicine. PMID- 17138963 TI - Nitric oxide signaling in oxytocin-mediated cardiomyogenesis. AB - Oxytocin (OT), a hormone recently identified in the heart, induces embryonic and cardiac somatic stem cells to differentiate into cardiomyocytes (CM), possibly through nitric oxide (NO). We verified this hypothesis using P19 cells and P19 Clone 6 derivatives expressing a green fluorescent protein (GFP) reporter linked to cardiac myosin light chain-2v promoter. OT treatment of these cells induced beating cell colonies that were fully inhibited by N,G-nitro-L-arginine-methyl ester (L-NAME), an inhibitor of NO synthases (NOS), partially reduced by 1400W, an inhibitor of inducible NOS, and ODQ, an inhibitor of NO-sensitive guanylyl cyclases. The NO generator S-nitroso-N-acetylpenicillamine (SNAP) reversed the L NAME inhibition of cell beating and GFP expression. In OT-induced cells, L-NAME significantly decreased transcripts of the cardiac markers Nkx2.5, MEF2c, alpha myosin heavy chain, and less, GATA4, endothelial NOS, and atrial natriuretic peptide, as well as the skeletal myocyte (SM) marker myogenin. Image analysis of OT-induced P19Cl6-GFP cells revealed ventricular CM coexpressing sarcomeric alpha actinin and GFP, with some cells exclusively expressing alpha-actinin, most likely of the SM phenotype. The OT-mediated production of CM, but not SM, was diminished by L-NAME. In P19 cells, exogenously added OT stimulated the expression of its own transcript, which was reduced in the presence of L-NAME. Surprisingly, L-NAME alone decreased the expression of anti-stage specific embryonic antigen-1 marker of the undifferentiated state and induced some beating colonies as well as GFP in P19Cl6-GFP cells. Collectively, our data suggest that the pleiotropic action of NO is involved in the initiation of CM differentiation of P19 cells and maintenance of their undifferentiated state. PMID- 17138964 TI - Antibody targeting of stem cells to infarcted myocardium. AB - Hematopoietic stem cell (HSC) therapy for myocardial repair is limited by the number of stem cells that migrate to, engraft in, and proliferate at sites of injured myocardium. To alleviate this limitation, we studied whether a strategy using a bispecific antibody (BiAb) could target human stem cells specifically to injured myocardium and preserve myocardial function. Using a xenogeneic rat model whereby ischemic injury was induced by transient ligation of the left anterior descending artery (LAD), we determined the ability of a bispecific antibody to target human CD34+ cells to specific antigens expressed in ischemic injured myocardium. A bispecific antibody comprising an anti-CD45 antibody recognizing the common leukocyte antigen found on HSCs and an antibody recognizing myosin light chain, an organ-specific injury antigen expressed by infarcted myocardium, was prepared by chemical conjugation. CD34+ cells armed and unarmed with this BiAb were injected intravenously in rats 2 days postmyocardial injury. Immunohistochemistry studies showed that the armed CD34+ cells specifically localized to the infarcted region of the heart, colocalized with troponin T stained cells, and colocalization with vascular structures. Compared to unarmed CD34+ cells, the bispecific antibody improved delivery of the stem cells to injured myocardium, and such targeted delivery was correlated with improved myocardial function 5 weeks after infarction (p < .01). Bispecific antibody targeting offers a unique means to improve the delivery of stem cells to facilitate organ repair and a tool to study stem cell biology. PMID- 17138965 TI - Flagellin-induced tolerance of the Toll-like receptor 5 signaling pathway in polarized intestinal epithelial cells. AB - Salmonella typhimurium is a gram-negative enteric pathogen that invades the mucosal epithelium and is associated with diarrheal illness in humans. Flagellin from S. typhimurium and other gram-negative bacteria has been shown to be the predominant proinflammatory mediator through activation of the basolateral Toll like receptor 5 (TLR5). Recent evidence has shown that prior exposure can render immune cells tolerant to subsequent challenges by TLR ligands. Accordingly, we examined whether prior exposure to purified flagellin would render human intestinal epithelial cells insensitive to future contact. We found that flagellin-induced tolerance is common to polarized epithelial cells and prevents further activation of proinflammatory signaling cascades by both purified flagellin and Salmonella bacteria but does not affect TNF-alpha stimulation of the same pathways. Flagellin tolerance is a rapid process that does not require protein synthesis, and that occurs within 1 to 2 h of flagellin exposure. Prolonged flagellin exposure blocks activation of the NF-kappaB, MAPK, and phosphoinositol 3-kinase signaling pathways and results in the internalization of a fraction of the basolateral TLR5 without affecting the polarity or total expression of TLR5. After removal of flagellin, cells require more than 24 h to fully recover their ability to mount a normal proinflammatory response. We have found that activation of phosphoinositol 3-kinase and Akt by flagellin has a small damping effect in the early stages of flagellin signaling but is not responsible for tolerance. Our study indicates that inhibition of TLR5-associated IL-1 receptor-associated kinase-4 activity occurs during the development of flagellin tolerance and is likely to be the cause of tolerance. PMID- 17138966 TI - Organized migration of epithelial cells requires control of adhesion and protrusion through Rho kinase effectors. AB - Migration of epithelial cell sheets, a process involving F-actin restructuring through Rho family GTPases, is both physiologically and pathophysiologically important. Our objective was to clarify the mechanisms whereby the downstream RhoA effector Rho-associated coil-coil-forming kinase (ROCK) influences coordinated epithelial cell motility. Although cells exposed to a pharmacological ROCK inhibitor (Y-27632) exhibited increased spreading in wound closure assays, they failed to migrate in a cohesive manner. Two main phenomena were implicated: the formation of aberrant protrusions at the migrating front and the basal accumulation of F-actin aggregates. Aggregates reflected increased membrane affiliation and detergent insolubility of the actin-binding protein ezrin and enhanced coassociation of ezrin with the membrane protein CD44. While F-actin aggregation following ROCK inhibition was recapitulated by inhibiting myosin light chain (MLC) phosphorylation with the MLC kinase inhibitor ML-7, the latter did not influence protrusiveness and, in fact, significantly decreased cell migration. Our results suggest that excessive protrusiveness downstream of ROCK inhibition reflects an influence of ROCK on F-actin stability via LIM kinase 1 (LIMK-1), which phosphorylates and inactivates cofilin. Y-27632 reduced the levels of both active LIMK-1 and inactive cofilin (phospho forms), and expression of a dominant negative LIMK-1 mutant stimulated leading edge protrusiveness. Furthermore, Y-27632-induced protrusions were partially reversed by overexpression of LIMK-1 to restore cofilin phosphorylation. In summary, our results provide new evidence suggesting that adhesive and protrusive events involved in organized epithelial motility downstream of ROCK are separately coordinated through the phosphorylation of (respectively) MLC and cofilin. PMID- 17138967 TI - The extracellular calcium-sensing receptor reciprocally regulates the secretion of BMP-2 and the BMP antagonist Noggin in colonic myofibroblasts. AB - To understand whether postprandial extracellular Ca(2+) (Ca(o)(2+)) changes were related to intestinal epithelial homeostasis, we performed array analysis on extracellular calcium-sensing receptor (CaSR)-expressing colonic myofibroblasts (18Co cells) and observed increases in bone morphogenetic protein (BMP)-2 transcripts. The present experiments demonstrated that regulated secretion of BMP 2 occurs in response to CaSR activation of these cells and revealed a new property of BMP-2 on the intestinal barrier. Activation by Ca(o)(2+), spermine, GdCl(3), or neomycin sulfate of 18Co cells or primary isolates of myofibroblasts from the normal human colon stimulated both the synthesis (RT-PCR) and secretion (ELISA) of BMP-2. Transient transfection with short interfering RNA against CaSR completely inhibited BMP-2 secretion. Transient transfection with dominant negative CaSR (R185Q) increased the EC(50) of Ca(o)(2+) (5.7 vs. 2.3 mM). Upregulation of BMP-2 transcript and secretion occurring within 3 h of CaSR activation was prevented by actinomycin D. CaSR-mediated BMP-2 synthesis and secretion required phosphatidylinositol 3-kinase activation (as assessed by phospho-Akt generation). Exogenous BMP-2 and conditioned medium from CaSR stimulated 18Co cells accelerated restitution in wounded postconfluent Caco-2 cells. Exogenous BMP-2 and conditioned medium from CaSR-stimulated 18Co cells increased the transepithelial resistance of low- and high-resistance T-84 epithelial monolayers. CaSR stimulation of T-84 epithelia and colonic myofibroblasts downregulated the BMP family antagonist Noggin, as assessed by RT PCR and Western blot analysis. Together, our data suggest that the CaSR mediates the effective concentration of BMP-2 in the intestine, which leads to enhanced repair and barrier development. PMID- 17138968 TI - Microinjection of exogenous somatostatin in the dorsal vagal complex inhibits pancreatic secretion via somatostatin receptor-2 in rats. AB - Previous studies have suggested that somatostatin inhibits pancreatic secretion at a central vagal site, and the dorsal vagal complex (DVC) is involved in central feedback inhibition of the exocrine pancreas. The aim of this study was to investigate the effect of exogenous somatostatin in the DVC on pancreatic secretion and the somatostatin receptor subtype(s) responsible for the effect. The effects of somatostatin microinjected into the DVC on pancreatic secretion stimulated by cholecystokinin octapeptide (CCK-8) or 2-deoxy-d-glucose (2-DG) were examined in anesthetized rats. To investigate the somatostatin inhibitory action site, a somatostatin receptor antagonist [SRA; cyclo(7-aminoheptanoyl-Phe d-Trp-Lys-Thr)] was microinjected into the DVC before intravenous infusion of somatostatin and CCK-8/2-DG. The effects of injection of a somatostatin receptor 2 agonist (seglitide) and combined injection of somatostatin and a somatostatin receptor-2 antagonist (CYN 154806) in the DVC on the pancreatic secretion were also investigated. Somatostatin injected into the DVC significantly inhibited pancreatic secretion evoked by CCK-8 or 2-DG in a dose-dependent manner. SRA injected into the DVC completely reversed the inhibitory effect of intravenous administration of somatostatin. Seglitide injected into the DVC also inhibited CCK-8/2-DG-induced pancreatic protein secretion. However, combined injection of somatostatin and CYN 154806 did not affect the CCK-8/2-DG-induced pancreatic secretion. Somatostatin in the DVC inhibits pancreatic secretion via somatostatin receptor-2, and the DVC is the action site of somatostatin for its inhibitory effect. PMID- 17138969 TI - Intestinal ribosomal p70(S6K) signaling is increased in piglet rotavirus enteritis. AB - Recent identification of the mammalian target of rapamycin (mTOR) pathway as an amino acid-sensing mechanism that regulates protein synthesis led us to investigate its role in rotavirus diarrhea. We hypothesized that malnutrition would reduce the jejunal protein synthetic rate and mTOR signaling via its target, ribosomal p70 S6 kinase (p70(S6K)). Newborn piglets were artificially fed from birth and infected with porcine rotavirus on day 5 of life. Study groups included infected (fully fed and 50% protein calorie malnourished) and noninfected fully fed controls. Initially, in "worst-case scenario studies," malnourished infected piglets were killed on days 1, 3, 5, and 11 postinoculation, and jejunal samples were compared with controls to determine the time course of injury and p70(S6K) activation. Using a 2 x 2 factorial design, we subsequently determined if infection and/or malnutrition affected mTOR activation on day 3. Western blot analysis and immunohistochemistry were used to measure total and phosphorylated p70(S6K); [(3)H]phenylalanine incorporation was used to measure protein synthesis; and lactase specific activity and villus-crypt dimensions were used to quantify injury. At the peak of diarrhea, the in vitro jejunal protein synthetic rate increased twofold (compared with the rate in the uninfected pig jejunum), concomitant with increased jejunal p70(S6K) phosphorylation (4-fold) and an increased p70(S6K) level (3-fold, P < 0.05). Malnutrition did not alter the magnitude of p70(S6K) activation. Immunolocalization revealed that infection produced a major induction of cytoplasmic p70(S6K) and nuclear phospho-p70(S6K), mainly in the crypt. A downregulation of semitendinosus muscle p70(S6K) phosphorylation was seen at days 1-3 postinoculation. In conclusion, intestinal activation of p70(S6K) was not inhibited by malnutrition but was strongly activated during an active state of mucosal regeneration. PMID- 17138970 TI - Loss of exocrine pancreatic stimulation during parenteral feeding suppresses digestive enzyme expression and induces Hsp70 expression. AB - Luminal nutrients are essential for the growth and maintenance of digestive tissue including the pancreas and small intestinal mucosa. Long-term loss of luminal nutrients such as during animal hibernation has been shown to result in mucosal atrophy and a corresponding stress response characterized by the induction of heat shock protein (Hsp)70 expression. This study was conducted to determine if the loss of luminal nutrients during total parenteral nutrition (TPN) would result in atrophy of the exocrine pancreas and small intestinal mucosa as well as an induction of Hsp70 expression in rats. In experiment 1, the treatment groups included an orally fed control, a saline-infused surgical control, or TPN treatment for 7 days. In experiment 2, the treatment groups included an orally fed control and TPN alone or coinfused with varying doses of glucagon-like peptide (GLP)-2, a mucosal proliferation agent, for 7 days. In experiment 1, TPN resulted in a 40% reduction in pancreatic mass that was associated with a dramatic reduction in digestive enzyme expression, enhanced apoptosis, and a 200% increase in Hsp70 expression. Conversely, heat shock cognate 70, Hsp27, and Hsp60 expression was not changed in the pancreas. In experiment 2, TPN resulted in a 30% reduction in jejunal mucosa mass and a similar induction of Hsp70 expression. The inclusion of GLP-2 during TPN attenuated jejunal mucosal atrophy and inhibited Hsp70 expression, suggesting that Hsp70 induction is sensitive to cell growth. These data indicate that pancreatic and intestinal mucosal atrophy caused by a loss of luminal nutrient stimulation is accompanied by a compensatory response involving Hsp70. PMID- 17138971 TI - Diagnosis using search engines. PMID- 17138972 TI - Sexual relationships between doctors and former patients. PMID- 17138973 TI - Donor breast milk banking. PMID- 17138974 TI - Reconfiguring acute hospitals in England. PMID- 17138975 TI - Former spy's death causes public health alert. PMID- 17138976 TI - Patients' views on access will affect GPs' pay. PMID- 17138977 TI - US cancer patients have problems with insurance and care, says survey. PMID- 17138980 TI - Abortion should be made easier, charity says. PMID- 17138981 TI - FDA approves silicone breast implants 14 years after their withdrawal. PMID- 17138987 TI - Bowel cancer treatment gets better, report says. PMID- 17138982 TI - Hungary to close 9000 hospital beds to bring it closer to EU average. PMID- 17138988 TI - Regulator finds advertising of complementary product "misleading". PMID- 17138990 TI - US health insurers propose expansion of cover, but plan is widely criticised. PMID- 17138995 TI - Feeling the squeeze. PMID- 17138996 TI - Rheumatic fever and its management. PMID- 17138997 TI - Misdiagnosis of angle closure glaucoma. PMID- 17138998 TI - ABC of obesity. Obesity and reproduction. PMID- 17138999 TI - Was message of sudden infant death study misleading? PMID- 17139000 TI - Does practice based commissioning avoid the problems of fundholding? PMID- 17139001 TI - Physical activity to prevent obesity in young children: dietary and behavioural modifications in managing childhood obesity. PMID- 17139002 TI - Physical activity to prevent obesity in young children: BMI in the BMJ. PMID- 17139003 TI - Physical activity to prevent obesity in young children: negative outcome or protocol problem? PMID- 17139004 TI - Methadone tolerance testing in drug misusers: beyond the edge of safety. PMID- 17139005 TI - Author's response to influenza vaccination: policy v evidence. PMID- 17139006 TI - Refusing to provide a prenatal test: can it ever be ethical? Rights of future children. PMID- 17139007 TI - Refusing to provide a prenatal test: can it ever be ethical? Time to re-think the autonomy of future individuals. PMID- 17139009 TI - Even clinician led management won't fix the NHS. PMID- 17139010 TI - Refusing to provide a prenatal test: can it ever be ethical? Ethics or humanity? PMID- 17139011 TI - Refusing to provide a prenatal test: can it ever be ethical? The darker side of medicine. PMID- 17139012 TI - Pedagogy and educational technologies of the future. PMID- 17139013 TI - Reshaping our educational experiences. PMID- 17139014 TI - APA summit on medical student education task force on informatics and technology: steps to enhance the use of technology in education through faculty development, funding and change management. AB - OBJECTIVE: This article provides an overview of how trainees, faculty, and institutions use technology for acquiring knowledge, skills, and attitudes for practicing modern medicine. METHOD: The authors reviewed the literature on medical education, technology, and change, and identify the key themes and make recommendations for implementing technology in medical education. RESULTS: Administrators and faculty should initially assess their own competencies with technology and then develop a variety of teaching methods that use technology to improve their curricula. Programs should decrease the general knowledge-based content of curricula and increase the use of technology for learning skills. For programs to be successful, they must address faculty development, change management, and funding. CONCLUSIONS: Willingness for change, collaboration, and leadership at all levels are essential factors for successfully implementing technology. PMID- 17139015 TI - The importance of distributed broadband networks to academic biomedical research and education programs. AB - OBJECTIVE: This article highlights the importance of distributed broadband networks as part of the core infrastructure necessary to deliver academic research and education programs. METHOD: The authors review recent developments in the field and present the University of California, Davis, environment as a case study of a future virtual regional academic medical center. RESULTS: Achieving the potential benefits of distributed broadband networks through regional health information organizations will involve considerable collaboration between different academic groups and the development and implementation of standardized technological and business approaches. CONCLUSIONS: Broadband technologies offer a substantial opportunity to improve the way academic education and research programs are delivered. PMID- 17139016 TI - Technology as an instrument to improve quality, accountability, and reflection in academic medicine. AB - OBJECTIVE: This article describes two complementary technology systems used in academic medicine to 1) improve the quality of learning and teaching, and 2) describe the barriers and obstacles encountered in implementing these systems. METHOD: The literature was integrated with in-depth, case-based experience with technology related to student progression, faculty promotion and school administration. RESULTS: Academic medicine concerns itself with data and outcomes. Psychiatrists need to attend to their learning and teaching paths as much as to developing the knowledge and skills to manage their patients. CONCLUSIONS: Technology enables us to track, manage, and report these data with increasing ease, making transparency and accuracy more achievable. PMID- 17139017 TI - Considerations in change management related to technology. AB - OBJECTIVE: The authors describe the complexity of social processes for implementing technological change. Once a new technology is available, information about its availability and benefits must be made available to the community of users, with opportunities to try the innovations and find them worthwhile, despite organizational resistances. METHOD: The authors reviewed the literature from psychiatry, psychology, sociology, business, and technology to distill common denominators for success and failure related to implementing technology. RESULTS: Beneficial technological innovations that are simple to use and obviously save everyone time and effort are easy to inaugurate. However, innovations that primarily serve management rather than subordinates or front line utilizers may fail, despite considerable institutional effort. This article reviews and outlines several of the more prominent theoretical models governing successful institutional change. CONCLUSIONS: Successful implementation of difficult technological changes requires visionary leadership that has carefully considered the benefits, consulted with influence leaders at all organizational levels to spot unintended consequences and sources of resistance, and developed a detailed plan and continuous quality assurance process to foster implementation over time. PMID- 17139018 TI - Students' and residents' perceptions regarding technology in medical training. AB - OBJECTIVE: This pilot study provides firsthand feedback from medical students and residents in training regarding their perceptions of technology in medicine. METHOD: The authors distributed an e-mail invitation to an anonymous Web-based survey to medical students and residents in two different U.S. training institutions. RESULTS: Respondents unanimously expressed that technology skills were important in medical training and felt it most important to learn about electronic medical records and accessing scientific information on the Internet. At the point of patient care, trainees' preferred reference sources were the Internet and PDA, in that order. Most clinical trainees felt PDAs were critical in patient care and met their clinical needs, and they were most likely to use them for medication reference. The majority of trainees preferred printed media over digital media for initial learning, but the converse for referencing. Instructor-led small groups were viewed as the best environment in which to receive instruction. CONCLUSIONS: Trainees in medical education are technologically savvy and provide invaluable feedback regarding initiation, development and refinement of technological systems in medical training. PMID- 17139019 TI - Visualizing the future: technology competency development in clinical medicine, and implications for medical education. AB - OBJECTIVE: In this article, the authors ask three questions. First, what will physicians need to know in order to be effective in the future? Second, what role will technology play in achieving that high level of effectiveness? Third, what specific skill sets will physicians need to master in order to become effective? METHOD: Through three case vignettes describing past, present, and potential future medical practices, the authors identify trends in major medical, technological and cultural shifts that will shape medical education and practice. RESULTS: From these cases, the authors generate a series of technology-related competencies and skill sets that physicians will need to remain leaders in the delivery of medical care. Physicians will choose how they will be end-users of technology, technology developers, and/or the interface between users and developers. These choices will guide the types of skills each physician will need to acquire. Finally, the authors explore the implications of these trends for medical educators, including the competencies that will be required of educators as they develop the medical curriculum. CONCLUSIONS: Examining historical and social trends, including how users adopt current and emerging technologies, allows us to anticipate changes in the practice of medicine. By considering market pressures, global trends and emerging technologies, medical educators and practicing physicians may prepare themselves for the changes likely to occur in the medical curriculum and in the marketplace. PMID- 17139020 TI - E-Learning systems: promises and pitfalls. AB - OBJECTIVE: This review examines the use of e-learning systems in psychiatric education. METHOD: The authors review the Instructional System Design process, focusing on ADDIE (Analysis, Design, Development, Implementation, and Evaluation). The authors also report on two representative teaching projects, discussing the planning, work, and assessment required. RESULTS: The basic operations of these education software systems are relatively easy to learn. The real work is the project preparation which precedes software use and final implementation. CONCLUSIONS: Internet-based learning software is another tool in the educator's armamentarium. Educators must still utilize sound educational pedagogy to create effective learning instruments. PMID- 17139021 TI - Online resources for assessment and evaluation. AB - OBJECTIVE: The Accreditation Council for Graduate Medical Education and the Liaison Committee on Medical Education have mandated the transition from written global evaluation methods to competency-based assessments in resident and medical student training. Assessment of competency requires analysis of performance data from numerous sources. This article reviews characteristics of Web-based evaluation and assessment systems and recommends areas for further development. METHOD: The authors review functions common to a variety of online evaluation and curriculum management systems with attention to their adaptation for competency documentation and assessment. Details of online global assessments, examination methods, electronic portfolios, procedure/case logs, and survey systems are provided along with a list of Internet resources. RESULTS: Online evaluation and assessment systems not only provide data on trainee competence but can provide valuable feedback on faculty teaching, curriculum quality, and the learning environment. Suggestions for future development, legal ambiguities that could become obstacles, and the need to clarify the responsibility for funding this work are discussed. Web-based performance assessment and evaluation are seen in the larger context of moving toward competency assessment of practicing physicians. CONCLUSIONS: Online assessment systems offer advantages over paper systems in allowing robust data analysis, reporting, and flexibility. A national online procedure/case log tracking system would facilitate gathering data that could provide one type of experiential benchmark for determining competency. PMID- 17139022 TI - Assessment of clinical skills using simulator technologies. AB - OBJECTIVE: Simulation technologies are used to assess and teach competencies through the provision of reproducible stimuli. They have exceptional utility in assessing responses to clinical stimuli that occur sporadically or infrequently. In this article, the authors describe the utility of emerging simulation technologies, and discuss critical issues in simulator-based skills assessment and appropriate results analysis. METHOD: Based on literature search and expert consensus, the authors discuss three simulation technologies: standardized patients and the objective structured clinical examination; the integrated high fidelity mannequin; virtual clinical stations and the objective structured virtual examination. RESULTS: The authors explore the current state of these technologies: uses, cost, limitations, and likely future applications. For instance, tele-standardized patients may test learners' communication/management approach to challenges during tele-consultation, such as a suicidal patient several hundred miles away. Integrated mannequins may test leadership skills during psychiatric emergencies. Case-based interactive virtual clinical assessment tools may test learners' decision-making skills or self-reflection. However, these exciting tools must be implemented systematically. Specifically, educators must define the competencies of interest precisely. Appropriate data analysis will generate dependable results, ascribing the correct proportion of outcome variability to individual learner behavior. Careful analysis and utilization of results will allow justification of the costs to major stakeholders. CONCLUSIONS: Simulation technologies offer exciting possibilities for skills evaluation and clinical practice improvement. When used creatively and appropriately, they form a useful adjunct in the armament of educators addressing the question, "Is this physician competent?" PMID- 17139023 TI - Personal digital assistants in psychiatric education. AB - OBJECTIVE: This article describes the various administrative and clinical applications for PDA use in psychiatric care and review the process for implementation in an academic medical center. METHOD: The authors reviewed the psychiatric literature and tested various hardware and software products. RESULTS: The literature describes various uses of the PDA as a reference tool, in clinical care, and in training documentation. CONCLUSIONS: The PDA is capable of numerous vital roles in medical education. PMID- 17139024 TI - Electronic medical records and their impact on resident and medical student education. AB - OBJECTIVE: Electronic medical records (EMRs) are becoming prevalent and integral tools for residents and medical students. EMRs can integrate point-of-service information delivery within the context of patient care. Though it may be an educational tool, little is known about how EMR technology is currently used for medical learners. METHOD: The authors reviewed the available published literature about the impact of EMRs on learners, including learners' attitudes about EMRs, educational uses of EMRs, and the potential effects of EMRs on learners' daily work. RESULTS: Research on EMRs for education is in its infancy. The authors found fewer than 50 articles with evidence on their use in medical education. The applications to education included point-of-care knowledge delivery, computerized clinical decision support systems, profiling of learner experiences, and daily workflow management. The evidence was mainly derived from single institution studies and occasionally across disciplines. CONCLUSIONS: EMRs have great potential as an educational tool, but thus far, strong data to support their use for this are lacking. As the usage of EMRs rises, educators must continue to study how best to use technology as an educational tool and as a tool to improve the daily work of residents and medical students. PMID- 17139025 TI - Virtual reality, telemedicine, web and data processing innovations in medical and psychiatric education and clinical care. AB - OBJECTIVE: This article highlights technology innovations in psychiatric and medical education, including applications from other fields. METHOD: The authors review the literature and poll educators and informatics faculty for novel programs relevant to psychiatric education. RESULTS: The introduction of new technologies requires skill at implementation and evaluation to assess the pros and cons. There is a significant body of literature regarding virtual reality and simulation, including assessment of outcomes, but other innovations are not well studied. CONCLUSIONS: Innovations, like other uses of technology, require collaboration between parties and integration within the educational framework of an institution. PMID- 17139026 TI - Education about hallucinations using an internet virtual reality system: a qualitative survey. AB - OBJECTIVE: The authors evaluate an Internet virtual reality technology as an education tool about the hallucinations of psychosis. METHOD: This is a pilot project using Second Life, an Internet-based virtual reality system, in which a virtual reality environment was constructed to simulate the auditory and visual hallucinations of two patients with schizophrenia. Eight hundred sixty-three self referred users took a self-guided tour. RESULTS: Five hundred seventy-nine (69%) of the users who toured the environment completed a survey. Of the survey responders, 440 (76%) thought the environment improved their understanding of auditory hallucinations, 69% thought it improved their understanding of visual hallucinations, and 82% said they would recommend the environment to a friend. CONCLUSIONS: Computer simulations of the perceptual phenomena of psychiatric illness are feasible with existing personal computer technology. Integration of the evaluation survey into the environment itself was possible. The use of Internet-connected graphics environments holds promise for public education about mental illness. PMID- 17139027 TI - Lifelong learning: skills and online resources. AB - OBJECTIVE: Advances in information technology enable the practicing psychiatrist's quest to keep up-to-date with new discoveries in psychiatry, as well as to meet recertification requirements. However, physicians' computer skills do not always keep up with technology, nor do they take advantage of online search and continuing education services. This article describes the rationale for using electronic databases and training, as well as basic computer skills, computer equipment, and important online resources for psychiatrists to meet their continuing education and recertification needs quickly, easily, and conveniently. METHOD: A literature review was performed using PUBMED and Google to find articles related to recertification, physician's technology adoption and computer skills, evidence-based medicine, and basic approaches to lifelong learning using computers and the Internet, and resources for lifelong learning. RESULTS: Psychiatrists are required to master a discrete set of information for board certification, and to maintain that knowledge for recertification. Surveys have shown that although most physicians use computers, the majority use them for personal or business purposes, and not for accessing Continuing Medical Education (CME) programs (1). The Council on Graduate Medical Education requires the acquisition of medical informatics skills for the Undergraduate Medical Education for the 21st Century (UME-21) curriculum project (2). There is a growing body of literature outlining basic computer skills and competencies for physicians to access online textbooks, databases, journals, and CME programs. CONCLUSIONS: Psychiatrists can benefit from learning how to use computers and the Internet to keep current with the advances in the field. Skills now being taught in medical school and residency are equally important for practicing psychiatrists to learn and master. PMID- 17139029 TI - Smoke-free legislation and mental health units: the challenges ahead. AB - Under the proposed English Health Act regulations most mental health units will have to be smoke-free, although patients will be able to smoke outside. Implementing the regulations will be challenging but may also be an opportunity for a more holistic approach focusing on the physical and psychological health of patients. PMID- 17139030 TI - Distress and fear disorders: an alternative empirically based taxonomy of the "mood" and "anxiety" disorders. AB - The nosological organisation of DSM-IV and ICD-10 does not capture the empirical structure of the mood and anxiety disorders. Instead, they form a broad group of "internalising" disorders with two subclasses: distress disorders and fear disorders. This empirical structure should form the basis for revised taxonomies in DSM-V and ICD-11. PMID- 17139031 TI - Collaborative care for depression in primary care. Making sense of a complex intervention: systematic review and meta-regression. AB - BACKGROUND: The management of depression in primary care is a significant issue for health services worldwide. "Collaborative care" interventions are effective, but little is known about which aspects of these complex interventions are essential. AIMS: To use meta-regression to identify "active ingredients" in collaborative care models for depression in primary care. METHOD: Studies were identified using systematic searches of electronic databases. The content of collaborative care interventions was coded, together with outcome data on antidepressant use and depressive symptoms. Meta-regression was used to examine relationships between intervention content and outcomes. RESULTS: There was no significant predictor of the effect of collaborative care on antidepressant use. Key predictors of depressive symptom outcomes included systematic identification of patients, professional background of staff and specialist supervision. CONCLUSIONS: Meta-regression may be useful in examining "active ingredients" in complex interventions in mental health. PMID- 17139032 TI - Treatment options in moderate and severe depression: decision analysis supporting a clinical guideline. AB - BACKGROUND: Treatment options for depression include antidepressants, psychological therapy and a combination of the two. AIMS: To develop cost effective clinical guidelines. METHOD: Systematic literature reviews were used to identify clinical, utility and cost data. A decision analysis was then conducted to compare the benefits and costs of antidepressants with combination therapy for moderate and severe depression in secondary care in the UK. RESULTS: Over the 15 month analysis period, combination therapy resulted in higher costs and an expected 0.16 increase per person in the probability of remission and no relapse compared with antidepressants. The cost per additional successfully treated patient was 4056 UK pounds (95% CI1400-18300); the cost per quality-adjusted life year gained was 5777 UK pounds (95% CI1900-33 800) for severe depression and 14 540 UK pounds (95% CI 4800-79 400) for moderate depression. CONCLUSIONS: Combination therapy is likely to be a cost-effective first-line secondary care treatment for severe depression. Its cost-effectiveness for moderate depression is more uncertain from current evidence. Targeted combination therapy could improve resource utilisation. PMID- 17139033 TI - Perinatal circumstances and risk of offspring suicide. Birth cohort study. AB - BACKGROUND: A higher risk of suicide has been associated with low birth weight in one study, but not yet replicated. Higher birth order has been associated with self-harm, but not with suicide. AIMS: To examine the relationship between perinatal circumstances and subsequent young adult suicide in Scotland. METHOD: Using linked data from the Scottish Morbidity Record and Scottish death records, a birth cohort of 1061830 people was followed-up for a mean of 25.1 years. Data were analysed using Cox regression. RESULTS: Higher maternal parity, younger maternal age (<25 years), non-professional parental occupations and low birth weight (<2500 g) were independently associated with higher suicide risk of offspring as young adults. There was no independent association with gestational age. CONCLUSIONS: Our findings provide support for the influence of maternal circumstance and foetal experience on subsequent mental health. PMID- 17139034 TI - Adherence therapy for people with schizophrenia. European multicentre randomised controlled trial. AB - BACKGROUND: There is equivocal evidence of the effectiveness of adherence therapy in improving treatment adherence and clinical outcomes for people with schizophrenia. AIMS: To evaluate the effectiveness of adherence therapy in improving quality of life for people with schizophrenia. METHOD: A 52-week, single-blind, multicentre randomised controlled trial of the effectiveness of adherence therapy. Participants were individually randomised to receive eight sessions of adherence therapy or health education. Assessments were undertaken at baseline and at 52-week follow-up. RESULTS: Adherence therapy was no more effective than health education in improving quality of life. CONCLUSIONS: This effectiveness trial provides evidence for the lack of effect of adherence therapy in people with schizophrenia with recent clinical instability, treated in ordinary clinical settings. PMID- 17139035 TI - Influence of sub-syndromal symptoms after remission from manic or mixed episodes. AB - BACKGROUND: Sub-syndromal symptoms in bipolar disorder impair functioning and diminish quality of life. AIMS: To examine factors associated with time spent with sub-syndromal symptoms and to characterise how these symptoms influence outcomes. METHOD: In a double-blind randomised maintenance trial, patients received either olanzapine or lithium monotherapy for 1 year. Stepwise logistic regression models were used to identify factors that were significant predictors of percentage time spent with sub-syndromal symptoms. The presence of sub syndromal symptoms during the first 8 weeks was examined as a predictor of subsequent relapse. RESULTS: Presence of sub-syndromal depressive symptoms during the first 8 weeks significantly increased the likelihood of depressive relapse (relative risk 4.67, P<0.001). Patients with psychotic features and those with a greater number of previous depressive episodes were more likely to experience sub syndromal depressive symptoms (RR=2.51, P<0.001 and RR=2.35, P=0.03 respectively). CONCLUSIONS: These findings help to identify patients at increased risk of affective relapse and suggest that appropriate therapeutic interventions should be considered even when syndromal-level symptoms are absent. PMID- 17139036 TI - Predicting community violence from patients discharged from mental health services. AB - BACKGROUND: The MacArthur Violence Risk Assessment Study (MacVRAS) in the USA provided strong evidence to support an actuarial approach in community violence risk assessment. AIMS: To examine the predictive accuracy of the MacVRAS measures, in addition to structured professional judgement, in a UK sample of patients discharged from in-patient care in the north-west of England. METHOD: A prospective study of 112 participants assessed pre-discharge and followed up at 24 weeks post-discharge. Pre-discharge measures were compared with prevalence of violent behaviour to determine predictive validity of risk factors. RESULTS: Historical measures of risk and measures of psychopathy, impulsiveness and anger were highly predictive of community violence. The more dynamic clinical and risk management factors derived from structured professional judgement (rated at discharge) added significant incremental validity to the historical factors in predicting community violence. CONCLUSIONS: Although static measures of risk relating to past history and personality make an important contribution to assessment of violence risk, consideration of current dynamic factors relating to illness and risk management significantly improves predictive accuracy. PMID- 17139037 TI - Group cognitive-behavioural therapy for schizophrenia. Randomised controlled trial. AB - BACKGROUND: The efficacy of cognitive-behavioural therapy for schizophrenia is established, but there is less evidence for a group format. AIMS: To evaluate the effectiveness of group cognitive-behavioural therapy for schizophrenia. METHOD: In all, 113 people with persistent positive symptoms of schizophrenia were assigned to receive group cognitive-behavioural therapy or treatment as usual. The primary outcome was positive symptom improvement on the Positive and Negative Syndrome Scales. Secondary outcome measures included symptoms, functioning, relapses, hopelessness and self-esteem. RESULTS: There were no significant differences between the cognitive-behavioural therapy and treatment as usual on measures of symptoms or functioning or relapse, but group cognitive-behavioural therapy treatment resulted in reductions in feelings of hopelessness and in low self-esteem. CONCLUSIONS: Although group cognitive-behavioural therapy may not be the optimum treatment method for reducing hallucinations and delusions, it may have important benefits, including feeling less negative about oneself and less hopeless for the future. PMID- 17139038 TI - Facial emotion processing in criminal psychopathy. Preliminary functional magnetic resonance imaging study. AB - BACKGROUND: It has been suggested that people with psychopathic disorders lack empathy because they have deficits in processing distress cues (e.g. fearful facial expressions). AIMS: To investigate brain function when individuals with psychopathy and a control group process facial emotion. METHOD: Using event related functional magnetic resonance imaging we compared six people scoring > or =25 on the Hare Psychopathy Checklist-Revised and nine non-psychopathic healthy volunteers during an implicit emotion processing task using fearful, happy and neutral faces. RESULTS: The psychopathy group showed significantly less activation than the control group in fusiform and extrastriate cortices when processing both facial emotions. However, emotion type affected response pattern. Both groups increased fusiform and extrastriate cortex activation when processing happy faces compared with neutral faces, but this increase was significantly smaller in the psychopathy group. In contrast, when processing fearful faces compared with neutral faces, the control group showed increased activation but the psychopathy group decreased activation in the fusiform gyrus. CONCLUSIONS: People with psychopathy have biological differences from controls when processing facial emotion, and the pattern of response differs according to emotion type. PMID- 17139039 TI - Structure of internalising symptoms in early adulthood. AB - BACKGROUND: Debate surrounds the underlying structure of internalising disorders including major depression, generalised anxiety disorder, phobias and panic disorders. AIMS: To model the within-time and across-time relationships of internalising symptoms, incorporating effects from generalised internalising and disorder-specific components of continuity. METHOD: Data were gathered from a 25 year longitudinal study of a birth cohort of 953 New Zealand children. Outcome measures included DSM-IV symptom scores for major depression, generalised anxiety disorder, phobia and panic disorder at the ages of 18, 21 and 25 years. RESULTS: Structural equation modelling showed that, within-times, a common underlying measure of generalised internalising explained symptom score comorbidities. Across-time correlation of symptom scores was primarily accounted for by continuity over time in generalised internalising. However, for major depression and phobia there was also evidence of across-time continuity in the disorder specific components of symptoms. CONCLUSIONS: Internalising symptoms can be partitioned into components reflecting both a generalised tendency to internalising and disorder-specific components. PMID- 17139040 TI - Risk factors for common mental disorders in women. Population-based longitudinal study. AB - BACKGROUND: The determinants of common mental disorders in women have not been described in longitudinal studies from a low-income country. METHOD: Population based cohort study of 2494 women aged 18 to 50 years, in India. The Revised Clinical Interview Schedule was used for the detection of common mental disorders. RESULTS: There were 39 incident cases of common mental disorder in 2166 participants eligible for analysis (12-month rate 1.8%, 95% CI 1.3-2.4%). The following baseline factors were independently associated with the risk for common mental disorder: poverty (low income and having difficulty making ends meet); being married as compared with being single; use of tobacco; experiencing abnormal vaginal discharge; reporting a chronic physical illness; and having higher psychological symptom scores at baseline. CONCLUSIONS: Programmes to reduce the burden of common mental disorder in women should target poorer women, women with chronic physical illness and who have gynaecological symptoms, and women who use tobacco. PMID- 17139041 TI - Long-term course of cognitive impairment in schizophrenia. AB - The aim of this study was to examine the long-term course of cognitive impairment in people with schizophrenia. Forty-three people with schizophrenia were followed up over an average of 33 years along with a control group of 12 matched individuals free from psychosis. Cognitive function was assessed at baseline and follow-up. The participants with schizophrenia were found to show impairment in verbal and non-verbal intelligence at baseline compared with estimated premorbid scores, this was not found in the control group. At follow-up there was a significant decline in non-verbal intelligence over time in participants with schizophrenia compared with controls (P=0.007). This differential change over time was not seen in verbal intelligence (P= 0.318). PMID- 17139042 TI - Delayed early proprioceptive information processing in schizophrenia. AB - It was first suggested that disordered proprioception was a core feature of schizophrenia by Sandor Rado in 1953. Using a recently designed proprioceptive event-related potential paradigm based on a change of load, we studied 12 unmedicated male out-patients with schizophrenia and 24 controls. In the patients, the early contralateral parietal activity was delayed and later central activity had increased amplitude, but gating was unaffected. The results could be understood within the "deficiency of corollary discharge" model of schizophrenia but not within the "filtering" theory. Further studies, including psychiatric controls, are necessary to verify the specificity of the abnormality. PMID- 17139043 TI - Processing facial emotions in adults with velo-cardio-facial syndrome: functional magnetic resonance imaging. AB - We studied the functional neuroanatomy of social behaviour in velo-cardio-facial syndrome (VCFS) using a facial emotional processing task and functional magnetic resonance imaging in adults with this syndrome and controls matched for age and IQ. The VCFS group had less activation in the right insula and frontal brain regions and more activation in occipital regions. Genetically determined abnormalities in pathways including those involved in emotional processing may underlie deficits in social cognition in people with VCFS. PMID- 17139044 TI - Urbanisation and the incidence of eating disorders. AB - The link between degree of urbanisation and a number of mental disorders is well established. Schizophrenia, psychosis and depression are known to occur more frequently in urban areas. In our primary care-based study of eating disorders, the incidence of bulimia nervosa showed a dose-response relation with degree of urbanisation and was five times higher in cities than in rural areas. Remarkably, anorexia nervosa showed no association with urbanisation. We conclude that urban life is a potential environmental risk factor for bulimia nervosa but not for anorexia nervosa. These findings provide a promising avenue for further research into the aetiology of eating disorders. PMID- 17139046 TI - Mental health and incapacity legislation. PMID- 17139047 TI - Mental health and incapacity legislation. PMID- 17139048 TI - Psychopathological consequences of ketamine. PMID- 17139050 TI - Acute psychotic presentations and acute psychosis. PMID- 17139051 TI - Moderate alcohol use and mental health. PMID- 17139052 TI - Stalking--a significant problem for patients and psychiatrists. PMID- 17139053 TI - Chronomics of suicides and the solar wind. PMID- 17139057 TI - Scientific Journals are "faith based": is there science behind peer review? PMID- 17139058 TI - Palpation of the skin--an important issue. PMID- 17139059 TI - Access to psychological and psychiatric services needs to be improved for the dying. PMID- 17139060 TI - Ways of seeing: biomedical perspectives on the social world. PMID- 17139061 TI - A brief history of homeopathy. PMID- 17139062 TI - "They would say that, wouldn't they?" A reader's guide to author and sponsor biases in clinical research. PMID- 17139063 TI - Jundi-Shapur, bimaristans, and the rise of academic medical centres. PMID- 17139064 TI - Dr Kahn's Museum: obscene anatomy in Victorian London. PMID- 17139065 TI - Attenti alle bufale ("beware of red herrings"), or, how to make evidence-based medicine work for you. PMID- 17139066 TI - In-flight emergencies: playing The Good Samaritan. PMID- 17139067 TI - How the doctor's nose has shortened over time; a historical overview of the truth telling debate in the doctor-patient relationship. PMID- 17139068 TI - Psychological services in hospices in the UK and Republic of Ireland. AB - OBJECTIVE: To evaluate the level of psychological services available to patients and staff in hospices. DESIGN: Questionnaire analysis. SETTING: Hospices in the UK and Republic of Ireland. PARTICIPANTS: 224 hospices. MAIN OUTCOME MEASURES: The availability of professional psychological support for those with advanced disease. RESULTS: Responses were received from 166 hospices (74%). Only 50 hospices (30%) have access to a psychiatrist, whilst 68 (41%) have access to a clinical psychologist and 92 (45%) have neither. Only 21 hospices (12%) have service level agreements with local mental health trusts. Counsellors, complementary therapists and spiritual advisors such as chaplains were more plentiful. CONCLUSIONS: Delivery of the NICE guidelines, especially tier four, may be compromised by limited availability of specialist services. This has implications for the psychological assessment of applicants for voluntary euthanasia under an Assisted Dying Act. PMID- 17139069 TI - The oldest surviving arthroplasty? PMID- 17139070 TI - Inventing the randomized double-blind trial: the Nuremberg salt test of 1835. PMID- 17139071 TI - Longer appointments for all! PMID- 17139072 TI - Private practice--definitely not a zero sum game. PMID- 17139073 TI - Poor handwriting remains a significant problem in medicine. PMID- 17139074 TI - Too little compassion in health care? PMID- 17139075 TI - Fear ever young: the terrorist death toll in perspective. PMID- 17139076 TI - Where has the humanity gone? PMID- 17139077 TI - Reasons to be cheerful (times three). PMID- 17139078 TI - Engineering the catalytic domain of human protein tyrosine phosphatase beta for structure-based drug discovery. AB - Protein tyrosine phosphatases (PTPs) play roles in many biological processes and are considered to be important targets for drug discovery. As inhibitor development has proven challenging, crystal structure-based design will be very helpful to advance inhibitor potency and selectivity. Successful application of protein crystallography to drug discovery heavily relies on high-quality crystal structures of the protein of interest complexed with pharmaceutically interesting ligands. It is very important to be able to produce protein-ligand crystals rapidly and reproducibly for as many ligands as necessary. This study details our efforts to engineer the catalytic domain of human protein tyrosine phosphatase beta (HPTPbeta-CD) with properties suitable for rapid-turnaround crystallography. Structures of apo HPTPbeta-CD and its complexes with several novel small-molecule inhibitors are presented here for the first time. PMID- 17139079 TI - The open-access high-throughput crystallization facility at EMBL Hamburg. AB - Here, the establishment of Europe's largest high-throughput crystallization facility with open access to the general user community is reported. The facility covers every step in the crystallization process from the preparation of crystallization cocktails for initial or customized screens to the setup of hanging-drop vapour-diffusion experiments and their automatic imaging. In its first year of operation, 43 internal and 40 external users submitted over 500 samples for a total of 2985 crystallization plates. An electronic booking system for registration, the selection of experimental parameters (e.g. drop size, sample-to-reservoir ratio) and the reservation of time slots was developed. External users can choose from more than 1000 initial crystallization conditions. By default, experiments are kept for six months and are imaged 15 times during this time period. A remote viewing system is available to inspect experiments via the internet. Over 100 stock solutions are available for users wishing to design customized screens. PMID- 17139080 TI - Structure of Mycobacterium tuberculosis thioredoxin C. AB - Mycobacterium tuberculosis is a facultative intracellular parasite of alveolar macrophages. M. tuberculosis is able to propagate in harsh environments within cells such as phagocytes, despite being exposed to reactive oxygen and nitrogen intermediates. The thioredoxin redox system is conserved across the phyla and has a well characterized role in resisting oxidative stress and influencing gene expression within prokaryotic and eukaryotic cells. M. tuberculosis thioredoxin (MtbTrx) has similar functions in redox homeostasis and it has recently been shown that alkyl hydroperoxidase C is efficiently reduced to its active form by MtbTrxC, supporting this notion. To address whether the MtbTrx has similar features to other thioredoxin structures and to examine the opportunities for designing drugs against this target, MtbTrxC has been crystallized and its structure determined to 1.3 A resolution. Unexpectedly, the structure demonstrates an interesting crystal packing in which five C-terminal residues from the MtbTrxC fold insert into a groove adjacent to the active site. A very similar interaction is observed in structures of human thioredoxins bound to peptides from the target proteins NF-kappaB and Ref-1. PMID- 17139081 TI - Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. AB - Gaucher disease is caused by mutations in the gene encoding acid-beta glucosidase. A recombinant form of this enzyme, Cerezyme, is used to treat Gaucher disease patients by ;enzyme-replacement therapy'. Crystals of Cerezyme after its partial deglycosylation were obtained earlier and the structure was solved to 2.0 A resolution [Dvir et al. (2003), EMBO Rep. 4, 704-709]. The crystal structure of unmodified Cerezyme is now reported, in which a substantial number of sugar residues bound to three asparagines via N-glycosylation could be visualized. The structure of intact fully glycosylated Cerezyme is virtually identical to that of the partially deglycosylated enzyme. However, the three loops at the entrance to the active site, which were previously observed in alternative conformations, display additional variability in their structures. Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. PMID- 17139082 TI - Structure of Escherichia coli UDP-N-acetylmuramoyl:L-alanine ligase (MurC). AB - The bacterial cell wall provides essential protection from the external environment and confers strength and rigidity to counteract internal osmotic pressure. Without this layer the cell would be easily ruptured and it is for this reason that biosynthetic pathways leading to the formation of peptidoglycan have for many years been a prime target for effective antibiotics. Central to this pathway are four similar ligase enzymes which add peptide groups to glycan moieties. As part of a program to better understand the structure-function relationships in these four enzymes, the crystal structure of Escherichia coli UDP-N-acetylmuramoyl:L-alanine ligase (MurC) has been determined to 2.6 A resolution. The structure was solved by multiwavelength anomalous diffraction methods from a single selenomethionine-substituted crystal and refined to a crystallographic R factor of 0.212 (R(free) = 0.259). The enzyme has a modular multi-domain structure very similar to those of other members of the mur family of ATP-dependent amide-bond ligases. Detailed comparison of these four enzymes shows that considerable conformational changes are possible. These changes, together with the recruitment of two different N-terminal domains, allow this family of enzymes to bind a substrate which is identical at one end and at the other has the growing peptide tail which will ultimately become part of the rigid bacterial cell wall. Comparison of the E. coli and Haemophilus influenzae structures and analysis of the sequences of known MurC enzymes indicate the presence of a ;dimerization' motif in almost 50% of the MurC enzymes and points to a highly conserved loop in domain 3 that may play a key role in amino-acid ligand specificity. PMID- 17139083 TI - What can be done with a good crystal and an accurate beamline? AB - X-ray single-wavelength anomalous diffraction (SAD) data from a crystal of proteinase K were collected using synchrotron radiation of 0.98 A wavelength at SER-CAT 22-ID beamline, Advanced Photon Source, Argonne National Laboratory. At this wavelength, the expected Bijvoet ratio resulting from the presence of one calcium, one chloride and ten S atoms in the 279-residue protein is extremely small at approximately 0.46%. The direct-methods program SHELXD located 11 anomalous sites using data truncated to 2 A resolution. SHELXE was used to produce an easily interpretable electron-density map. This study shows that an accurate beamline and a good-quality crystal provide the possibility of successfully using a very weak anomalous signal of sulfur measured at a short wavelength for phasing a protein structure, even if a small degree of radiation damage is present. PMID- 17139084 TI - High-resolution structure of the p53 core domain: implications for binding small molecule stabilizing compounds. AB - The p53 transcriptional regulator is the most frequently mutated protein in human cancers and the majority of tumor-derived p53 mutations map to the central DNA binding core domain, with a subset of these mutations resulting in reduced p53 stability. Here, the 1.55 A crystal structure of the mouse p53 core domain with a molecule of tris(hydroxymethyl)aminomethane (Tris) bound through multiple hydrogen bonds to a region of p53 shown to be important for repair of a subset of tumor-derived p53-stability mutations is reported. Consistent with the hypothesis that Tris binding stabilizes the p53 core domain, equilibrium denaturation experiments are presented that demonstrate that Tris binding increases the thermodynamic stability of the mouse p53 core domain by 3.1 kJ mol(-1) and molecular-dynamic simulations are presented revealing an overall reduction in root-mean-square deviations of the core domain of 0.7 A when Tris is bound. It is also shown that these crystals of the p53 core domain are suitable for the multiple-solvent crystal structure approach to identify other potential binding sites for possible core-domain stabilization compounds. Analysis of the residue specific temperature factors of the high-resolution core-domain structure, coupled with a comparison with other core-domain structures, also reveals that the L1, H1-S5 and S7-S8 core-domain loops, also shown to mediate various p53 activities, harbor inherent flexibility, suggesting that these regions might be targets for other p53-stabilizing compounds. Together, these studies provide a molecular scaffold for the structure-based design of p53-stabilization compounds for development as possible therapeutic agents. PMID- 17139085 TI - The 1.8 A resolution structure of hydroxycinnamoyl-coenzyme A hydratase-lyase (HCHL) from Pseudomonas fluorescens, an enzyme that catalyses the transformation of feruloyl-coenzyme A to vanillin. AB - The crystal structure of hydroxycinnamoyl-CoA hydratase-lyase (HCHL) from Pseudomonas fluorescens AN103 has been solved to 1.8 A resolution. HCHL is a member of the crotonase superfamily and catalyses the hydration of the acyl-CoA thioester of ferulic acid [3-(4-hydroxy-3-methoxy-phenyl)prop-2-enoic acid] and the subsequent retro-aldol cleavage of the hydrated intermediate to yield vanillin (4-hydroxy-3-methoxy-benzaldehyde). The structure contains 12 molecules in the asymmetric unit, in which HCHL assumes a hexameric structure of two stacked trimers. The substrate, feruloyl-CoA, was modelled into the active site based on the structure of enoyl-CoA hydratase bound to the feruloyl-CoA-like substrate 4-(N,N-dimethylamino)-cinnamoyl-CoA (PDB code 1ey3). Feruloyl-CoA was bound in this model between helix 3 of the A subunit and helix 9 of the B subunit. A highly ordered structural water in the HCHL structure coincided with the thioester carbonyl of feruloyl-CoA in the model, suggesting that the oxyanion hole for stabilization of a thioester-derived enolate, characteristic of coenzyme A dependent members of the crotonase superfamily, is conserved. The model also suggested that a strong hydrogen bond between the phenolic hydroxyl groups of feruloyl-CoA and BTyr239 may be an important determinant of the enzyme's ability to discriminate between the natural substrate and cinnamoyl-CoA, which is not a substrate. PMID- 17139086 TI - Structure of the UNC5H2 death domain. AB - UNC5Hs (UNC5H1-4) are netrin 1 receptors that are involved in axonal guidance and neuronal migration. They are dependence receptors that mediate apoptosis in the absence of netrin 1. UNC5H2-induced apoptosis depends on the interaction of the death domain at the C-terminus with the DAP-kinase death domain and caspase cleavage near the transmembrane region. Here, the crystal structure of the mouse UNC5H2 death domain has been determined at 2.1 A resolution. The domain adopts a six-helix bundle fold, which is similar to those of the other members of the death-domain superfamily. The UNC5H2 death domain is a dimer in the crystal and in solution. This homodimerized structure may represent the structure of the death domain when netrin 1 binds to the UNC5H2 receptor. Homodimerization of UNC5H2 may block the access of caspase to the cleavage site. In the death-domain dimer, residues in alpha3 and the 3(10)-helix preceding alpha3 and the residues in alpha4 make significant contacts, mainly by hydrophobic and van der Waals interactions. PMID- 17139087 TI - Structures of the interacting domains from yeast glutamyl-tRNA synthetase and tRNA-aminoacylation and nuclear-export cofactor Arc1p reveal a novel function for an old fold. AB - Eukaryotic aminoacyl-tRNA synthetases (aaRS) frequently contain additional appended domains that are absent from their prokaryotic counterparts which mediate complex formation between eukaryotic aaRS and cofactors of aminoacylation and translation. However, the structural basis of such interactions has remained elusive. The heteromerization domain of yeast glutamyl-tRNA synthetase (GluRS) has been cloned, expressed, purified and crystallized in space group C222(1), with unit-cell parameters a = 52, b = 107, c = 168 A. Phase information was obtained from multiple-wavelength anomalous dispersion with selenomethionine to 2.5 A resolution and the structure, comprising two monomers per asymmetric unit, was determined and refined to 1.9 A resolution. The structure of the interacting domain of its accessory protein Arc1p was determined and refined to 1.9 A resolution in a crystal form containing 20 monomers organized in five tetramers per asymmetric unit (space group C2, unit-cell parameters a = 222, b = 89, c = 127 A, beta = 99.4 degrees ). Both domains adopt a GST-like fold, demonstrating a novel role for this fold as a protein-protein interaction module. PMID- 17139088 TI - Analysis of protein hydration in ultrahigh-resolution structures of the SRP GTPase Ffh. AB - Two new structures of the SRP GTPase Ffh have been determined at 1.1 A resolution and provide the basis for comparative examination of the extensive water structure of the apo conformation of these GTPases. A set of well defined water binding positions have been identified in the active site of the two-domain ;NG' GTPase, as well as at two functionally important interfaces. The water hydrogen bonding network accommodates alternate conformations of the protein side chains by undergoing local rearrangements and, in one case, illustrates binding of a solute molecule within the active site by displacement of water molecules without further disruption of the water-interaction network. A subset of the water positions are well defined in several lower resolution structures, including those of different nucleotide-binding states; these appear to function in maintaining the protein structure. Consistent arrangements of surface water between three different ultrahigh-resolution structures provide a framework for beginning to understand how local water structure contributes to protein-ligand and protein-protein binding in the SRP GTPases. PMID- 17139089 TI - Ultrahigh-resolution study of protein atomic displacement parameters at cryotemperatures obtained with a helium cryostat. AB - Two X-ray data sets for a complex of human aldose reductase (h-AR) with the inhibitor IDD 594 and the cofactor NADP(+) were collected from two different parts of the same crystal to a resolution of 0.81 A at 15 and 60 K using cold helium gas as cryogen. The contribution of temperature to the atomic B values was estimated by comparison of the independently refined models. It was found that although being slightly different for different kinds of atoms, the differences (deltaB) in the isotropic equivalents B of atomic displacement parameters (ADPs) were approximately constant (about 1.7 A(2)) for well ordered atoms as the temperature was increased from 15 to 60 K. The mean value of this difference varied according to the number of non-H atoms covalently bound to the parent atom. Atoms having a B value of higher than 8 A(2) at 15 K showed much larger deviations of deltaB from the average value, which might reflect partial occupancy of atomic sites. An analysis of the anisotropy of ADPs for individual atoms revealed an increase in the isotropy of ADPs with the increase of the temperature from 15 to 60 K. In a separate experiment, a 0.93 A resolution data set was collected from a different crystal of the same complex at 100 K using cold nitrogen as a cryogen. The effects of various errors on the atomic B values were estimated by comparison of the refined models and the temperature-dependent component was inferred. It was found that both decreasing the data redundancy and increasing the resolution cutoff led to an approximately constant increase in atomic B values for well ordered atoms. PMID- 17139090 TI - Structure of the ribosomal protein L1-mRNA complex at 2.1 A resolution: common features of crystal packing of L1-RNA complexes. AB - The crystal structure of a hybrid complex between the bacterial ribosomal protein L1 from Thermus thermophilus and a Methanococcus vannielii mRNA fragment containing an L1-binding site was determined at 2.1 A resolution. It was found that all polar atoms involved in conserved protein-RNA hydrogen bonds have high values of density in the electron-density map and that their hydrogen-bonding capacity is fully realised through interactions with protein atoms, water molecules and K(+) ions. Intermolecular contacts were thoroughly analyzed in the present crystals and in crystals of previously determined L1-RNA complexes. It was shown that extension of the RNA helices providing canonical helix stacking between open-open or open-closed ends of RNA fragments is a common feature of these and all known crystals of complexes between ribosomal proteins and RNAs. In addition, the overwhelming majority of complexes between ribosomal proteins and RNA molecules display crystal contacts formed by the central parts of the RNA fragments. These contacts are often very extensive and strong and it is proposed that they are formed in the saturated solution prior to crystal formation. PMID- 17139091 TI - X-ray diffraction structure of a cell-wall invertase from Arabidopsis thaliana. AB - Cell-wall invertases play crucial roles during plant development. They hydrolyse sucrose into its fructose and glucose subunits by cleavage of the alpha1-beta2 glycosidic bond. Here, the structure of the Arabidopsis thaliana cell-wall invertase 1 (AtcwINV1; gene accession code At3g13790) is described at a resolution of 2.15 A. The structure comprises an N-terminal fivefold beta propeller domain followed by a C-terminal domain formed by two beta-sheets. The active site is positioned in the fivefold beta-propeller domain, containing the nucleophile Asp23 and the acid/base catalyst Glu203 of the double-displacement enzymatic reaction. The function of the C-terminal domain remains unknown. Unlike in other GH 32 family enzyme structures known to date, in AtcwINV1 the cleft formed between both domains is blocked by Asn299-linked carbohydrates. A preliminary site-directed mutagenesis experiment (Asn299Asp) removed the glycosyl chain but did not alter the activity profile of the enzyme. PMID- 17139092 TI - Molecular replacement in the 'twilight zone': structure determination of the non haem iron oxygenase NovR from Streptomyces spheroides through repeated density modification of a poor molecular-replacement solution. AB - Crystals of recombinant NovR (subunit MW = 29 924 Da; 270 amino acids), a non haem iron oxygenase from Streptomyces spheroides, were grown by vapour diffusion. The protein crystallized in space group C2, with unit-cell parameters a = 86.69, b = 139.38, c = 100.82 A, beta = 101.18 degrees . Native data were collected to a resolution of 2.1 A from a single crystal at a synchrotron and a molecular replacement solution was obtained using the program AMoRe. The starting phase information was very poor and did not permit model building. Phases were subsequently improved using a combination of fourfold averaging and very gradual phase extension in the program DM to yield an interpretable map. NovR belongs to a novel class of non-haem iron oxygenases that share sequence similarity with class II aldolases. It is predicted to perform two consecutive oxidative decarboxylation steps in the biosynthesis of the prenylated hydroxybenzoic acid moiety of the aminocoumarin antibiotic novobiocin. PMID- 17139094 TI - Stability indicating methods for the determination of some fluoroquinolones in the presence of their decarboxylated degrades. AB - Two stability-indicating methods, namely densitometric TLC and derivative spectrophotometry for the determination of the fluoroquinolone antibacterials lomefloxacin (Lfx), moxifloxacin (Mfx), and sparfloxacin (Sfx) in the presence of their acid degrades are described. Acid degradation was adopted and the main decarboxylated product separated by TLC. Degradation products were identified confirming a previously mentioned degradation scheme. The densitometric method is based on the separation of the intact drug from its acid degradation product on silica gel G plates using different mobile phases and the spots of the intact drugs were scanned at 288, 290, and 292 nm for Lfx, Mfx, and Sfx, respectively. The derivative spectrophotometric method utilizes first derivative D(1) UV spectrophotometry with zero crossing points at 295.2 nm for Lfx, 280.4 and 303.4 nm for Mfx, and 280.8 nm for Sfx. Regression analysis of Beer's plots showed good correlation in the concentration ranges 0.2-1.2, 0.1-1.4, and 0.5-2.0 microg/spot for Lfx, Mfx, and Sfx, respectively, in the densitometric method and 2-16 microg/ml for all drugs in the derivative spectrophotometric method. The proposed methods were successfully applied for the determination of the investigated drugs in bulk powder with mean percentage accuracy ranges from 98.93 to 101.25% for the TLC method and from 98.18 to 100.35% for the D(1) method. The proposed methods were also applied for the determination of the investigated drugs in their pharmaceutical dosage forms and their validity was assessed using the standard addition technique with mean percentage recovery ranging from 100.25 to 101.70% in the TLC method and from 99.27 to 102.12% in the D(1) method. The selectivity of the proposed methods was tested by the analysis of laboratory-prepared mixtures containing different percentages of the studied drugs and their acid degrades. The proposed methods were found selective for the determination of the intact drugs in the presence of up to 90% of their degrades in the TLC method and 70% for Lfx and 90% for Mfx and Sfx in the D(1) method. PMID- 17139095 TI - Stereo structure-controlled and electronic structure-controlled estrogen-like chemicals to design and develop non-estrogenic bisphenol A analogs based on chemical hardness concept. AB - The aim of this study was to elucidate the structure-activity relationship of bisphenol A (BPA) analogs using absolute hardness (eta) and absolute electronegativity (chi) (chemical hardness) and to design a non-estrogen active BPA. To determine the structure-activity relationships of BPA analogs, we investigated MCF-7 cell proliferation stimulated by BPA analogs and an eta-chi diagram based on the electronic structure of the BPA analogs. The results show that the actions of the environmental hormones BPA analogs have two chemical properties; (i) 'stereo structure-controlled' and (ii) 'electronic structure controlled' estrogen-like chemical activities. Therefore, we designed and synthesized BPA analogs which do not possess these 2 characteristics, ((i) and (ii)), and demonstrate the non-estrogen activity of the analog. PMID- 17139096 TI - Two new dioxopiperazine derivatives, arestrictins A and B, isolated from Aspergillus restrictus and Aspergillus penicilloides. AB - In the course of searching for bioactive substances, two new dioxopiperazine derivatives, arestrictins A (1) and B (2), were isolated along with the known dioxopiperazine, cristatin A (3), and the known peptide, asperglucide (4), from the organic extract of the xerophilic fungi, Aspergillus restrictus and Aspergillus penicilloides. The absolute structures of 1 and 2, except for the configuration of the secondary alcohol in 1, were established by spectroscopic and chemical investigation. The absolute configuration of cristatin A (3) was also determined. PMID- 17139097 TI - Spectrophotometric determination of fluoxetine by batch and flow injection methods. AB - A rapid, simple, and accurate spectrophotometric method is presented for the determination of fluoxetine by batch and flow injection analysis methods. The method is based on fluoxetine competitive complexation reaction with phenolphthalein-beta-cyclodextrin (PHP-beta-CD) inclusion complex. The increase in the absorbance of the solution at 554 nm by the addition of fluoxetine was measured. The formation constant for fluoxetin-beta-CD was calculated by non linear least squares fitting. Fluoxetine can be determined in the range 7.0 x 10( 6)-2.4 x 10(-4) mol l(-1) and 5.0 x 10(-5)-1.0 x 10(-2) mol l(-1) by batch and flow methods, respectively. The limit of detection and limit of quantification were respectively 4.13 x 10(-6) mol l(-1) and 1.38 x 10(-5) mol l(-1) for batch and 2.46 x 10(-5) mol l(-1) and 8.22 x 10(-5) mol l(-1) for flow method. The sampling rate in flow injection analysis method was 80+/-5 samples h(-1). The method was applied to the determination of fluoxetine in pharmaceutical formulations and after addition to human urine samples. PMID- 17139098 TI - Isolation and structure of four new ceramides from the starfish Luidia maculata. AB - A new sphingosine-type ceramide LMCer-1-1 (1) and three new phytosphingosine-type ceramides, LMCer-2-1 (2), LMCer-2-6 (3), and LMCer-2-7 (4), were isolated from the anti-hyperglycemic active ceramide molecular species LMCer-1 and LMCer-2, obtained from the less polar fraction of the chloroform-methanol extract of the whole bodies of Luidia maculata. The structures of these ceramides were determined on the basis of chemical and spectroscopic evidence as: (2S,3R,4E,2'R) 2-(2-hydroxyhexadecanoylamino)-16-methyl-4-octadecene-1,3-diol (1), (2S,3S,4R,2'R)-2-(2-hydroxyhexadecanoylamino)-16-methyl-octadecane-1,3,4-triol (2), (2S,3S,4R,2'R)-2-(2-hydroxydocosanoylamino)-hexadecane-1,3,4-triol (3), and (2S,3S,4R,2'R)-2-(2-hydroxydocosanoylamino)-14-methyl-hexadecane-1,3,4-triol (4). PMID- 17139099 TI - Two new glycosides from the soft coral Sinularia firma. AB - Two new glycosides, named Firmacosides A and B, together with known fatty esters, batyl alcohol, Delta(5,20) sterol and sphingosine derivatives have been isolated from the soft coral Sinularia firma TIX-DUR. On the basis of spectroscopic analysis ((1)H-NMR, (13)C-NMR, (1)H-(1)H COSY, HMQC, HMBC and FAB-MS), Firmacoside A was established as hexadecanyl-1-O-alpha-D-arabinopyranosyloxy (1- >4)-alpha-D-arabinopyranosyloxy (1-->4)-alpha-D-arabinopyranoside (1), and firmacoside B was elucidated as docosanyl-1-O-alpha-D-arabinopyranosyloxy (1-->4) 3-O-acetyl-alpha-arabinopyranosyloxy (1-->4)-alpha-D-arabinopyranoside (2). PMID- 17139100 TI - Synthesis of heterocyclic compounds via nucleophilic aroylation catalyzed by imidazolidenyl carbene. AB - Xanthones and acridones were synthesized from 3,4-difluoronitrobenzene and 2 fluorobenzaldehydes in two or three steps. The key step was nucleophilic aroylation catalyzed by imidazolidenyl carbene. The nucleophilic aroylation of 3,4-difluoronitrobenzene afforded 2,2'-difluoro-4-nitrobenzophenones. The cyclization of the difluorobenzophenones with O-nucleophile and N-nucleophile yielded 3-nitroxanthones and 3-nitroacridones, respectively. Indazole, quinolino[2,3-b]quinoxaline, and thianaphtho[2,3-b]quinoxaline derivatives were also synthesized via nucleophilic aroylation of 2,3-dichloroquinoxaline followed by cyclization with nucleophiles. PMID- 17139101 TI - Fungal citridone D having a novel phenylfuropyridine skeleton. AB - Citridone D was isolated from the culture broth of Penicillium sp. FKI-1938 by solvent extraction, silica gel column chromatography and HPLC. The structure of citridone D was elucidated by spectroscopic analysis including NMR analysis. Citridone D was found to have a novel phenylfuropyridine skeleton different from those of other citridones. Citridone D potentiated miconazole activity against Candida albicans. PMID- 17139102 TI - Biomimetic synthesis of (+/-)-galanthamine and asymmetric synthesis of (-) galanthamine using remote asymmetric induction. AB - (+/-)-Galanthamine (1) was synthesized in excellent yield by applying PIFA mediated oxidative phenol coupling of N-(4-hydroxy)phenethyl-N-(3',4',5' trialkoxy)benzyl formamide (15b) as a key step. Because of the symmetrical characteristics of the pyrogallol moiety in the substrate (15b), the phenol coupling resulted in a sole coupling product except for volatile components from the oxidizing agent. On the basis of the successful results of the above strategy, (-)-galanthamine (1) was synthesized by employing a novel remote asymmetric induction, where conformation of the seven-membered ring in the product of the phenol coupling was restricted by forming a fused-chiral imidazolidinone ring with D-phenylalanine on the benzylic C-N bond of the tri-O alkylated gallyl amino moiety. The conformational restriction and successive debenzylation of the protected hydroxyl groups on the pyrogallol ring caused diastereoselective cyclization to yield a cyclic ether having the desired stereochemistry for the synthesis of (-)-1. PMID- 17139103 TI - Starting point to molecular design: efficient automated 3D model builder Key3D. AB - Obtaining three-dimensional (3D) structures from structural formulae is a crucial process in molecular design. We have developed a new 3D model builder, Key3D, in which the simplified distance geometry technique and structure optimization based on the MMFF force field are combined. In an evaluation study using 598 crystal structures, the high performance and accuracy of Key3D were demonstrated. In the "flexible-fitting" test, which is focused on practical usefulness in the molecular design process, 88% of the Key3D structures acceptably reproduced the reference crystal structures (root-mean-square deviation <0.6 A) upon rotation of acyclic bonds. These results indicate that Key3D will be very effective in providing starting points for practical molecular design. PMID- 17139104 TI - Synthesis of 1,4-diphenylbutadiene derivatives: novel inducer of tissue-type plasminogen activator (t-PA) in cultured bovine endothelial cells. AB - (E,E)-1,4-Diphenylbutadiene derivatives were synthesized by utilizing the Stobbe reaction of dimethyl succinate as a key step. Their stereoisomers were also synthesized stereoselectively by means of the cross-coupling reaction of the vinylstannanes and the vinylbromides, which were obtained from the propiolic acid esters by stereoselective hydrostannation, as a key step. To discover novel stimulators of fibrinolysis in vascular endothelial cells, the synthesized compounds were added to cultured bovine endothelial cells to determine the activity of the plasminogen activator in the conditioned medium. Of the synthesized compounds, three compounds were found to stimulate the activity of the plasminogen activator in endothelial cells. In addtition, these compounds inhibited thrombus formation in a rat model of venous thrombosis. PMID- 17139105 TI - Antiproliferative constituents from umbelliferae plants. IX. New triterpenoid glycosides from the fruits of Bupleurum rotundifolium. AB - The MeOH extract of the fruits of Bupleurum rotundifolium showed inhibitory activity against human gastric adenocarcinoma (MK-1) cell growth. Bioactivity guided fractionation of the MeOH extract led to the isolation of four new triglycosides of 13beta,28-epoxy oleanane-type triterpenes, named rotundiosides O, Q, S and T; 12 new glycosides of oleanane-type triterpenes, named rotundiosides J-N, P, R, U-Y, and others; echinocystic acid 3-O-sulfate; and three known oleanane-type triterpene glycosides, rotundiosides A, F and G. The structures of the new isolates were determined based on chemical and spectroscopic evidence. The GI(50) of isolates against MK-1, HeLa and B16F10 cell lines are reported. PMID- 17139106 TI - Five new phenylethanoid glycosides from the whole plants of Lamium purpureum L. AB - Five new phenylethanoid glycosides, lamiusides A (1), B (2), C (3), D (4) and E (5), were isolated from the whole plants of Lamium purpureum L. (Labiatae) together with seven known compounds (6-12). On the basis of chemical and spectral analyses, the structures of the new compounds were elucidated to be 2-(3,4 dihydroxyphenyl)ethyl-O-beta-D-galactopyranosyl-(1-->2)-alpha-L-rhamnopyranosyl (1-->3)-(4-O-trans-caffeoyl)-beta-D-glucopyranoside (1), 2-(3,4 dihydroxyphenyl)ethyl-O-beta-D-galactopyranosyl-(1-->2)-alpha-L-rhamnopyranosyl (1-->3)-(4-O-trans-feruloyl)-beta-D-glucopyranoside (2), 2-(3,4 dihydroxyphenyl)ethyl-O-beta-D-galactopyranosyl-(1-->2)-alpha-L-rhamnopyranosyl (1-->3)-(6-O-trans-caffeoyl)-beta-D-glucopyranoside (3), 2-(3,4-dihydroxyphenyl) R,S-methoxy-ethyl-O-beta-D-galactopyranosyl-(1-->2)-alpha-L-rhamnopyranosyl-(1- >3)-(4-O-trans-caffeoyl)-beta-D-glucopyranoside (4) and 2-(3-hydroxy-4 methoxyphenyl)ethyl-O-alpha-L-rhamnopyranosyl-(1-->3)-beta-D-glucopyranosyl-(1- >6)-(4-O-cis-feruloyl)-beta-D-glucopyranoside (5). In addition, the radical scavenging activities of compounds 1-4 on 1,1-diphenyl-2-picrylhydrazyl radical were examined. PMID- 17139107 TI - Mono- and dihydroxylated metabolites of thalidomide: synthesis and TNF-alpha production-inhibitory activity. AB - Mono- and dihydroxylated metabolites of thalidomide were efficiently prepared and characterized, and their inhibitory activity on tumor necrosis factor (TNF)-alpha production in the human monocytic leukemia cell line THP-1 was evaluated. 5,N Dihydroxythalidomide was a much more potent TNF-alpha production inhibitor than thalidomide. PMID- 17139108 TI - Chemistry of unprotected amino acids in aqueous solution: direct bromination of aromatic amino acids with bromoisocyanuric acid sodium salt under strong acidic condition. AB - Brominations of unprotected aromatic amino acids such as phenylalanine, tyrosine, and glycine, with bromoisocyanuric acid mono sodium salt (BICA-Na) were conducted in 60% aq. H(2)SO(4) at 0 degrees C to give a mixture of mono-brominated products in good yield. Unexpectedly, meta-bromophenylglycine was obtained as main product accompanied by ortho- and para-substituted products, while phenylalanine gave only ortho- and para-substituted products. Bromination of 2-phenylethylamine or benzylamine showed a tendency similar to the corresponding amino acids. PMID- 17139109 TI - A new citryl glycoside from Gastrodia elata and its inhibitory activity on GABA transaminase. AB - A new citryl glycoside, trimethylcitryl-beta-D-galactopyranoside (1) along with a known phenolic compound, gastrodigenin (2) have been isolated from the active fraction of the rhizomes of Gastrodia elata (Orchidaceae). Their structures were elucidated on the basis of spectroscopic data and chemical reaction. 1 inhibited GABA transaminase activity by 56.8% at the final concentration of 10 microg/ml. PMID- 17139110 TI - The permeation of nalmefene hydrochloride across different regions of ovine nasal mucosa. AB - The permeability of nalmefene hydrochloride (NH) across different regions of ovine nasal mucosa was investigated in vitro. Five different regions of ovine nasal mucosa (superior turbinate mucosa, middle turbinate mucosa, inferior turbinate mucosa, posterior septum mucosa, and anterior septum mucosa) were studied. The results showed that the permeability coefficients of NH through different regions of nasal mucosa were different, and the suitable regions for the absorption of NH were the middle turbinate mucosa, the posterior septum mucosa and the superior turbinate. At the same time, the middle turbinate mucosa was the largest region among the five regions, thus it was the main absorption region for NH. The high uniformity of the middle turbinate mucosa also made it the most suitable model for the permeation of NH in vitro. PMID- 17139111 TI - Synthetic applications of Baylis-Hillman chemistry: an efficient and solely stereoselective synthesis of (E)-alpha-methylcinnamic acids and potent hypolipidemic agent LK-903 from unmodified Baylis-Hillman adducts. AB - An efficient and solely stereoselective synthesis of (E)-alpha-methylcinnamic acids has been accomplished in single pot by reduction of the unmodified Baylis Hillman adducts, methyl-3-hydroxy-3-aryl-2-methylenepropanoates with I(2)/NaBH(4) reagent system at room temperature followed by hydrolysis. The efficacy of this method has been proved in the total synthesis of 1-[p-(myristyloxy)-alpha methylcinnamoyl]glycerol, LK-903, a highly active hypolipidemic agent. PMID- 17139112 TI - Two diterpene rhamnosides, mimosasides B and C, from Mimosa hostilis. AB - Two new diterpene rhamnosides, mimosasides B and C (1, 2) were isolated together with mimosaside A (3), a known diterpene rhamnoside (4), four known flavones (5 8), five known flavanones (9-13), and four known chalcones (14-17) from the leaves and twigs of a Brazilian medicinal plant, Mimosa hostilis. PMID- 17139113 TI - Isolation of tetraprenyltoluquinols from the brown alga Sargassum thunbergii. AB - Thunbergols A (4) and B (5), tetraprenyltoluquinols, along with three known compounds (1-3) have been isolated from the brown alga Sargassum thunbergii. The structures of these two new compounds were determined to be 9-(3,4-dihydro-2,8 dimethyl-6-hydroxy-2H-1-benzopyran-2-yl)-6-methyl-2-(4-methyl-3-pentenyl)-(2E,6E) nonadienoic acid (4) and 10-(2,3-dihydro-5-hydroxy-7-methyl-1-benzofuran-2-yl)-10 hydroxy-6-methyl-2-(4-methyl-3-pentenyl)-(2E,6E)-undecadienoic acid (5), respectively, by combined spectroscopic methods. Both of them exhibited significant scavenging activities on radical and potently inhibited generation of ONOO(-) from morpholinosydnonimine (SIN-1). PMID- 17139114 TI - A new synthesis of 2-sulfanyl allylic alcohols and alpha-sulfanyl ketones from carbonyl compounds and 1-chloroalkyl p-tolyl sulfoxides with carbon-carbon bond formation. AB - Reaction of lithium alpha-sulfinyl carbanions of 1-chloroalkyl p-tolyl sulfoxides with ketones or aldehydes at low temperature gave adducts in almost quantitative yields. Treatment of the adducts derived from ketones with trifluoroacetic anhydride (TFAA) in the presence of NaI in acetone gave alpha-sulfanyl allylic alcohols in good to quantitative yields. On the other hand, treatment of the adducts derived from aldehydes with TFAA and NaI resulted in the formation of alpha-sulfanyl ketones and/or alpha-sulfanyl allylic alcohols. These reactions offer a good method for the synthesis of the above-mentioned compounds from ketones and aldehydes with carbon-carbon bond-formation in two steps and in good yields. PMID- 17139115 TI - Two new triterpenoid saponins isolated from Polygala japonica. AB - Bioassay guided investigation of whole parts of Polygala japonica afforded two new triterpenoid saponins, characterized as 3-O-beta-D-glucopyranosyl medicagenic acid 28-O-{beta-D-xylopyranosyl(1-->4)-[beta-D-apiofuranosyl(1-->3)]-alpha-L rhamnopyranosyl(1-->2)-beta-D-glucopyranosyl} ester (1), 3-O-beta-D glucopyranosyl 2-oxo-olean-12-en-23, 28-dioic acid 28-O-{beta-D-xylopyranosyl(1- >4)-[beta-D-apiofuranosyl(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-beta-D glucopyranosyl} ester (2), together with four known triterpenoid saponins (3-6). Their structures were elucidated by spectroscopic and chemical methods. Saponins 3, 4 and 5 showed significant anti-inflammation effects on carrageenan-induced acute paw edema in mouse. PMID- 17139116 TI - Asymmetric construction of quaternary carbon stereocenter by Pd-catalyzed intramolecular alpha-arylation. AB - The catalyst comprised of Pd(OAc)(2) and H(8)-BINAP provides good reaction conversions for a catalytic enantioselective intramolecular alpha-arylation of N (2-bromophenyl)-N-methyl-2-arylpropanamide to form the quaternary carbon with up to 68% enantioselectivity. PMID- 17139117 TI - The coronary circulation in adults with congenital heart disease. PMID- 17139118 TI - Screening for suspected chronic obstructive pulmonary disease with an eleven-item pre-interview questionnaire (11-Q). AB - OBJECTIVE: Early diagnosis is central to the management of chronic obstructive pulmonary disease (COPD). In time-constrained clinical situations, a pre interview questionnaire can be a useful method for alerting both clinicians and patients with COPD, particularly for elderly patients. To screen subjects who might have COPD, we have developed an efficient pre-interview questionnaire. METHODS: In study I, we developed an 11-item questionnaire (11-Q) to alert primary care providers to the possibility of COPD, and the validity of this questionnaire was investigated. In study II, a randomized and prospective cross sectional study was performed on a total of 245 subjects with or without respiratory symptoms. RESULTS: Good test-retest reliability and validity were shown in study I; the internal consistency of 11-Q was highly preserved (Cronbach alpha value of 0.867), with a high reproducibility of responses by inter- and intra examinees. Study II showed that the 11-Q in COPD patients with more than moderate severity was significantly higher than that in bronchial asthma or non cardiopulmonary subjects (both, p<0.0001). Among the COPD patients, the total score significantly distinguished the severity of COPD as mild or more than moderate. CONCLUSION: The pre-interview questionnaire, 11-Q, was found to be a useful tool to alert primary care providers to subjects with COPD and could also be used to distinguish COPD with a more than moderate severity from bronchial asthma. The 11-Q can be used as a simple and inexpensive method of predicting COPD, thus being a useful tool to alert primary care providers to patients with suspected COPD, particularly among the elderly. PMID- 17139119 TI - Adult meningism and viral meningitis, 1997-2004: clinical data and cerebrospinal fluid cytokines. AB - OBJECTIVE: Although meningism manifesting acute headache has been observed to be associated with common viral and bacterial infections, its definition and pathogenesis have not been clarified. Clinical findings and cerebrospinal fluid (CSF) cytokines in adult patients with meningism were investigated and compared with those in viral meningitis. PATIENTS AND METHODS: Among the adult inpatients in our hospital from 1997 to 2004, 5 with meningism and 17 with viral meningitis were identified according to the criteria described in this study, and their clinical data were analyzed. In the CSF samples of the 5 patients with meningism and the 17 with viral meningitis, the concentrations of interferon-gamma (IFN gamma), tumor necrosis factor-alpha (TNF-alpha), interleukin-2 (IL-2), IL-4, IL 6, and IL-10 were determined using a cytometric bead array. RESULTS: The five patients with meningism all showed fever and meningeal signs such as severe headache and nuchal stiffness without CSF pleocytosis (<5 cells/mm3). Four patients were associated with herpetic Kaposi's eczema, herpes simplex, or herpes zoster, and all five patients had favorable outcomes. The levels of all CSF cytokines in patients with meningism were below normal values, whereas IFN-gamma and IL-6 in patients with viral meningitis were moderately elevated. CONCLUSION: The normal cytokine levels in meningism may possibly reflect the lack of direct viral infection and may be helpful in differentiating both meningism and viral meningitis at an early stage. PMID- 17139120 TI - Successful treatment with adefovir of one patient whose cryoglobulinemic vasculitis relapsed under lamivudine therapy and who was diagnosed to have HBV virologic breakthrough with YMDD mutations. AB - We report a patient whose cryoglobulinemic vasculitis recurred due to reactivation of lamivudine-resistant HBV. Our patient with hepatitis B-related cryoglobulinemic vasculitis was administered lamivudine. Her vasculitis regressed, ALT normalized, HBV-DNA became negative. Under lamivudine therapy, her cryoglobulinemic cutaneous vasculitis recurred. ALT increased significantly; it was found that tyrosine-methionine-aspartate-aspartate (YMDD) motif in the DNA polymerase gene had been replaced by YIDD. Adefovir was added to lamivudine. During follow-up, her purpura disappeared, ALT normalized, HBV-DNA became negative. Our patient is the first whose cryoglobulinemic vasculitis recurred under lamivudine, who had a HBV virologic breakthrough with YMDD mutation, and was successfully treated with adefovir. PMID- 17139121 TI - A recovery case of acute-onset autoimmune hepatitis presenting as fulminant hepatic failure [corrected], who received living donor-liver transplantation. AB - A 23-year-old woman was admitted to our hospital with jaundice and hepatic coma. She had taken a weight-loss supplement for one month before admission. Her clinical and laboratory findings were consistent with fulminant hepatic failure and fulfilled the criteria of autoimmune hepatitis. Despite corticosteroid pulse therapy and plasma exchange, her symptoms and laboratory findings deteriorated. Her condition improved after she received a living donor-liver transplant from her sister. Autoimmune hepatitis usually follows a chronic course, but it should be considered a type of fulminant hepatic failure and treated promptly. PMID- 17139122 TI - Left ventricular pseudoaneurysm following acute myocardial infarction. AB - We describe a case of 57-year-old man who presented with acute myocardial infarction (AMI) and heart failure with rapid progression of cardiomegaly. Cardiac multislice computed tomography and echocardiography showed the ventricular pseudoaneurysm, probably due to cardiac free wall rupture caused by AMI. Cardiac CT is another useful tool for the non-invasive diagnosis of cardiac rupture. PMID- 17139123 TI - Acute liver dysfunction complicated with uncontrollable glycemia due to insulin antibody: successful treatment with glucocorticoid and lispro insulin. AB - Here, we report a case of acute liver dysfunction complicated with uncontrollable glycemia due to insulin antibody. The patient was admitted to our hospital due to diabetic ketoacidosis. He was administered insulin immediately, however, his fasting plasma glucose level remained unstable despite the insulin treatment. Blood biochemistry revealed severe liver dysfunction, although no markers including hepatitis virus or autoantibodies associated with autoimmune liver diseases were detected. The 125I-insulin binding rate was high (54%). The characteristics of insulin antibody in this patient were similar to the antibodies of IAS patients, therefore we administered oral glucocorticoid against insulin antibody. The reduction in the 125I-insulin binding rate and the binding capacity of the high affinity site of insulin antibodies were balanced after oral glucocorticoid therapy. In addition, preprandial subcutaneous regular insulin was switched to lispro insulin. Postprandial plasma glucose levels were relatively improved by lispro insulin. The etiology of acute liver dysfunction was unknown, however, we believe that the combination of oral glucocorticoid and lispro insulin was suitable and useful for preventing recurrent liver dysfunction in this patient. PMID- 17139124 TI - Subclinical Cushings disease with amelioration of metabolic comorbidities after removal of pituitary tumor. AB - A 49-year-old woman with hypertension, obesity and impaired glucose tolerance (IGT) was admitted for evaluation of pituitary incidentaloma. Although she presented no Cushingoid feature, endocrine examination of hypothalamo-pituitary adrenal (HPA) axis showed elevated basal plasma ACTH and cortisol levels, their lack of circadian rhythm, non-suppressibility to low-dose (1 mg) dexamethasone, and responsiveness to CRH, suggesting autonomous ACTH secretion from a pituitary tumor. She underwent transsphenoidal surgery, and was diagnosed as chromophobe adenoma with positive ACTH immunoreactivity. Postoperatively, her abnormal HPA axis was resolved, along with improvement of hypertension, obesity and IGT. Thus, her metabolic comorbidities are likely due to subclinical Cushings disease. PMID- 17139125 TI - Overt diabetes mellitus in a patient with combined primary aldosteronism and Cushing's syndrome. AB - Although there are some case reports of combined aldosterone and cortisol producing adrenal tumor, overt diabetes mellitus has been rarely described. A 55 year-old hypertensive woman had hypokalemia and overt hyperglycemia without Cushingoid clinical features. The body mass index was 18.2 kg/m2, fasting blood glucose was 302 mg/dl and hemoglobin A1c was 11.6%. Endogenous insulin secretion was well preserved, whereas insulin sensitivity measured by short insulin tolerance test was markedly impaired. A solitary left aldosterone- and cortisol producing adrenal tumor was diagnosed. We described a rare case of overt diabetes mellitus in a patient with combined primary aldosteronism and Cushing's syndrome. PMID- 17139126 TI - IgG4-associated multifocal systemic fibrosis complicating sclerosing sialadenitis, hypophysitis, and retroperitoneal fibrosis, but lacking pancreatic involvement. AB - A 71-year-old man was admitted with malaise, mild fever, anorexia, body weight loss, lower back pain, thirst, and polydipsia. He showed bilateral swelling of the submandibular glands. Examinations showed panhypopituitarism and a high serum IgG4 concentration. Fluorodeoxyglucose positron emission tomography (FDG-PET) revealed uptake in the pituitary gland, bilateral submandibular gland, bilateral hilar and mediastinal lymph nodes, and a mass consistent with retroperitoneal fibrosis, but not in the pancreas. Biopsy specimens from the submandibular gland and retroperitoneal mass indicated sialadenitis and retroperitoneal fibrosis respectively, and showed severe fibrosis and inflammation with marked lymphoplasmacytic infiltration and IgG4-positive plasma cell infiltration. Hormone replacement therapy with hydrocortisone resulted in marked clinical improvement. Systemic involvement found in this patient possibly corresponded to the new concept of IgG4-associated multifocal systemic fibrosis. PMID- 17139127 TI - Autoimmune hypophysitis treated with intravenous glucocorticoid therapy. AB - A 46-year-old man presented with frontal headache, a visual field defect and general fatigue. Magnetic resonance imaging (MRI) of the brain showed symmetrical enlargement of the pituitary gland and stalk due to the presence of a mass lesion extending toward the optic chiasm. Gadolinium injection further revealed homogeneous strong enhancement with involvement of the adjacent dura (dural tail). Basal plasma levels of ACTH, free thyroxine and gonadotropins were decreased, and 24-h urinary 17-OHCS excretion was reduced. An elevated anti thyroglobulin antibody titer indicated the presence of autoimmune thyroiditis. Under the suspicion of autoimmune hypophysitis, 60 mg/day prednisolone sodium succinate was intravenously administered for two weeks followed by a decreasing dose of oral prednisolone. Clinical symptoms and pituitary dysfunction recovered during steroid treatment and MRI showed marked shrinkage of the pituitary mass. Early initiation of an intravenous dose of glucocorticoid followed by oral steroid administration therefore seems to be an efficient treatment for autoimmune hypophysitis even in patients with visual dysfunction. PMID- 17139128 TI - Pneumonia and sepsis caused by Gemella morbillorum: an unusual association. PMID- 17139129 TI - Metastatic malignant melanoma mimicking primary lung adenocarcinoma. PMID- 17139130 TI - Homonymous hemianopia due to localized hemorrhage of the lateral geniculate body. PMID- 17139132 TI - Wernicke-like aphasia due to Broca area lesion. PMID- 17139131 TI - "Superior rectus paralysis" due to inferior rectus myxedema. PMID- 17139133 TI - Delivery after transfer of frozen-thawed embryos from in vitro-matured oocytes in a woman at risk for ovarian hyperstimulation syndrome. AB - The present report describes the birth of a healthy infant after cryopreservation of embryos produced from in vitro-matured oocytes retrieved from a woman at risk of developing ovarian hyperstimulation syndrome (OHSS) during conventional in vitro fertilization (IVF) cycles. A conventional long protocol including gonadotropin-releasing hormone agonist (GnRHa) and gonadotropins induced a risk of OHSS. Oocyte retrieval was performed on day 11 of the cycle, and 27 immature oocytes were obtained. Following incubation for 24 h in maturation medium, 74.1% (20/27) of the oocytes were at the metaphase II stage. Fourteen oocytes (14/20, 70.0%) were fertilized after intracytoplasmic sperm injection (ICSI) with her husband's spermatozoa and cultured for 3 days. On day 4 following oocyte retrieval, three embryos at the 8-16 cell stage were transferred into the woman's uterus, and five spare embryos were frozen. Since the fresh embryo transfer failed to result in pregnancy, three post-thaw embryos were transferred into the woman three months later. Transfer of the frozen embryos resulted in pregnancy with delivery of a healthy infant girl. PMID- 17139134 TI - Action of IGF-I on expression of the long form of the leptin receptor (ObRb) in the prepubertal period and throughout the estrous cycle in the mature pig ovary. AB - Leptin is an important satiety hormone and reproductive regulator found, along with its receptors, throughout the ovary. To date, the changes in ovarian expression of leptin receptor during the prepubertal period and throughout the estrous cycle in the pig have not been studied. In this study, the long form of the leptin receptor was a detectable level in immature pig ovarian follicles when assayed using semiquantitative reverse transcription-polimerase chain reaction (RT-PCR). Moreover, its level was increased in growing follicles during follicular phase of the estrous cycle (6-fold in early antral [EAF] and antral [AF] follicles) and was highest in newly formed corpora lutea. Its changes paralleled those in steroid secretion and especially progesterone (P(4)) secretion. Additionally, we showed that insulin-like growth factor (IGF)-I had stimulatory effect on leptin receptor expression using a tissue culture model of follicular and luteal cells, with a 12-fold increase in prepubertal ovaries and a 1.5- to 2-fold increase in growing follicles and newly formed corpora lutea (CL). These results suggest that ovarian expression of leptin receptor is regulated throughout the estrous cycle by ovarian steroids, with peak expression at ovulation, indicating a possible involvement in follicular development and corpus luteum formation. Moreover, this data points to an important role of IGF-I in leptin receptor expression during the entire estrous cycle, with a special role during the prepubertal period. PMID- 17139135 TI - Effect of recombinant bovine somatotropin (rbST) on cytoplasmic maturation of bovine oocytes and their developmental competence in vitro. AB - The objectives of this study were to evaluate the effects of recombinant bovine somatotropin (rbST) on the nuclear and cytoplasmic maturation of bovine oocytes and their further developmental competence to blastocysts in vitro. We analyzed the mitochondrial activity and concentration of intracellular stored calcium ([Ca(2+)](is)) in matured oocytes and the morphology and chromatin status of produced embryos after in vitro fertilization. Cumulus-oocyte complexes were incubated in TCM 199 containing 10% fetal calf serum (control medium 1: CM 1) or 10% estrus cow serum (control medium 2: CM 2). The culture medium of the treatment groups was modified by supplementation of the control medium with 10 ng/ml rbST (CM 1A and CM 2A), 10(6)/ml granulosa cells (CM 1B and CM 2B), or 10 ng/ml rbST plus 10(6)/ml granulosa cells (CM 1C and CM 2C). No differences were observed in the percentages of oocytes reaching metaphase II between the groups. However, the proportion of blastocysts was highest in treatment groups CM 1C and CM 2C (P<0.05). The type of serum did not alter the positive effect of rbST on the developmental competence of embryos. The fluorescence intensity of metabolically active mitochondria measured by intensity per oocyte (Em 570) after MitoTracker CMTM Ros Orange labeling was significantly increased in oocytes matured in the presence of 10 ng/ml rbST and granulosa cells (309.21 vs. 119.97 microA; P<0.01). In parallel, the concentration of [Ca(2+)](is) in oocytes, determined using fluorophore chlortetracycline, was significantly decreased (0.85 +/- 0.02 vs. 0.97 +/- 0.03 AU; P<0.05). Based on these results, we concluded that rbST, in interaction with granulosa cells stimulates the oxidative activity of ooplasmic mitochondria and decreases the content of [Ca(2+)](is) in oocytes. These facts support the hypothesis that somatotropin influences the developmental competence of bovine oocytes during maturation in vitro, and this effect can be modulated by granulosa cells. PMID- 17139136 TI - Developmental competence of cat oocytes from ovaries stored at various temperature for 24 h. AB - We conducted two experiments to investigate the effects of storage temperature and period for cat ovaries on the meiotic and developmental competence of oocytes collected from the ovaries. In Experiment 1, ovaries were stored in physiological saline for 24 h at 4 C, 23-25 C, or 38 C (cold, room, and incubator temperature groups, respectively), and then oocytes were collected from the ovaries in each group. Morphologically intact oocytes were then selected and cultured in maturation medium for 24 h. Significantly more oocytes reached metaphase II (MII) in the cold temperature group (53.4%) than in the room and incubator temperature groups (20.0 and 2.4%, respectively). In Experiment 2, ovaries were stored in physiological saline at room temperature for 0, 6, 12 or 18 h, and then they were stored at 4 C (cold storage) until reaching a total storage period of 24 h. After storage of the ovaries, morphologically intact oocytes were matured, fertilized with frozen-thawed spermatozoa, and cultured in vitro. The rates of morphologically intact oocytes obtained from the ovaries stored at room temperature for 0, 6, 12 or 18 h were 35.3, 30.0, 26.4 and 14.7%, respectively, and the rates of intact oocytes that reached MII were 63.2, 36.4, 26.5 and 11.9%, respectively. The results suggested that the numbers of morphologically intact oocytes and intact oocytes that reached MII after in vitro maturation decrease gradually as the period of storage at room temperature before cold storage increases. Only oocytes from ovaries stored for 6 h developed to the blastocyst stage after in vitro maturation and fertilization when ovaries were stored at room temperature before cold storage. These results indicate that 24 h storage of ovaries at high temperatures (>23 C) decreases the meiotic competence of oocytes. The quality and developmental competence of oocytes are influenced by the length of storage at room temperature before cold storage. PMID- 17139137 TI - Analysis of hyperpolarized 129Xe dynamics in mouse lungs under spontaneous respiration for separate determination of lung functional parameters and relaxation time. AB - Magnetic resonance spectroscopy (MRS) was used to investigate the dynamics of hyperpolarized (HP) 129Xe respiration in the chests of mice under spontaneous respiration. The washout curve was analyzed using Kety's exchange model of inert gases, and the 3 factors that affect the slope of the washout curve, i.e., the RF flip angle, respiratory parameters, and apparent relaxation time (which comprises terms including the relaxation time in alveoli, T1air, and perfusion), were determined separately. Flip angle was determined precisely using the dual flip angle method, and ventilation volume was determined using SF6 gas at thermal equilibrium. Furthermore, an attempt was made to separate out the terms of T1air and perfusion from the apparent relaxation time after exploiting the ventilation model of lungs in steady state. Values of relaxation time T1air=30.5 s and perfusion term lambdaQ/VA=0.016 s-1 were obtained, supporting the applicability of the ventilation model proposed. PMID- 17139138 TI - The effect of simultaneous use of respiratory triggering in diffusion-weighted imaging of the liver. AB - OBJECTIVE: To determine the effect of simultaneous use of respiratory triggering in hepatic diffusion-weighted imaging (DWI), we compared DWI with respiratory triggering (RT-DWI) and DWI under free breathing (FB-DWI) in terms of relative contrast between hepatic tumor and surrounding liver parenchyma, apparent diffusion coefficient (ADC) measurement, and frequency of respiratory misregistration. MATERIALS AND METHODS: Thirty patients (21 men, 9 women, aged 25 to 80 years) with liver metastasis or hepatocellular carcinoma in the right hepatic lobe were examined with RT- and FB-DWI. In patients having multiple tumors, up to 3 lesions were selected by mutual agreement of 2 diagnostic radiologists. Finally, 59 nodules were selected for evaluation. Relative contrast ratio (RCR) between the lesions and surrounding parenchyma and ADC were measured in each hepatic lesion in each sequence. The differences in RCR and ADC between RT- and FB-DWI were statistically analyzed using Wilcoxon's signed rank test. The frequency of respiratory misregistration advents in each sequence was visually evaluated by the 2 diagnostic radiologists and assessed using McNemar's test. RESULTS: RCRs were significantly higher in RT- than in FB-DWI (P<0.001). The difference in ADC between the 2 sequences was not significant. However, ADCs tended to be more scattered in FB- than in RT-DWI. The frequency of respiratory misregistration advents was significantly higher in FB- than in RT-DWI. CONCLUSION: FB- was inferior to RT-DWI in both contrast between tumors and surrounding liver parenchyma and in accuracy of ADC measurement as far as the numbers of excitation in each sequence were the same. For preoperative examination of hepatic resection, RT- is more suitable than FB-DWI. PMID- 17139139 TI - Dynamic high-spatial-resolution MR imaging of invasive ductal carcinoma: influence of histological scirrhous component on MR descriptors. AB - PURPOSE: The purpose of this study was to assess the relationship between the amount of scirrhous component in invasive ductal carcinoma and its MR characteristics. MATERIALS AND METHODS: We retrospectively reviewed 71 consecutive patients with invasive ductal carcinoma smaller than 25 mm (average, 16.6 mm) in diameter. The scirrhous component was defined as invasive foci in small clusters of cancer cells showing desmoplasia. Invasive ductal carcinoma was subclassified into 3 groups in accordance with the amount of the scirrhous component (scirrhous component degree; SCD): SCD I (scirrhous component less than 20%), SCD II (intermediate), and SCD III (more than 80%). Dynamic magnetic resonance (MR) imaging was performed using volumetric interpolated sequence. Prior to dynamic study, T2*-weighted first-pass perfusion images were obtained before, during, and after bolus injection of 0.1 mmol Gd-DTPA/kg. RESULTS: Twenty eight lesions were classified as SCD I, 14 as SCD II, and 29 as SCD III. Mass margin and signal intensity loss in the perfusion study were significantly different among the 3 SCD groups (P<0.001). The kinetic patterns were significantly different among the 3 SCD groups (P=0.04), and between SCD I/II and SCD III (P=0.03). The presence of enhancing internal septations was significantly different between SCD I/II and SCD III carcinomas (P=0.05). Central enhancement was only observed in SCD I carcinoma (4%; 3/71). CONCLUSION: The histological predominance of the scirrhous component in invasive ductal carcinoma may be one explanation for the differences in morphologic and kinetic patterns on MR imaging. PMID- 17139140 TI - Air microbubbles as MR susceptibility contrast agent at 1.5 Tesla. AB - Air microbubbles have been investigated recently at high magnetic field strength (2 Tesla or greater) as potential MR susceptibility contrast agents. We used a phantom to measure their susceptibility at 1.5 T to clarify their usefulness for this purpose. The phantom, filled with fresh Levovist suspension at 4 different doses (67 to 125 mg/mL), was continuously scanned with a gradient-echo technique at a temporal resolution of 10 s. The transverse relaxation increase (R2*) by microbubbles demonstrated a time course of exponential decay at each dose (time constant, 39 to 57 s). The dependency of R2* on microbubble volume fraction was linear, with a slope of 89 s-1 per percentage microbubble volume fraction. Our study represents the first step towards applying microbubbles as susceptibility contrast agents at 1.5 T. PMID- 17139141 TI - Prompt contrast enhancement of cerebrospinal fluid space in the fundus of the internal auditory canal: observations in patients with meningeal diseases on 3D FLAIR images at 3 Tesla. AB - We speculated that meningeal pathologies might facilitate the permeability of cranial nerves at the fundus of the internal auditory canal (IAC), causing prompt enhancement after administration of Gd-DTPA. Using a 3D- fluid-attenuated inversion recovery (FLAIR) sequence, we evaluated the enhancement of the cerebrospinal fluid (CSF) space in the IAC fundus 10 min after Gd-DTPA administration in patients with meningeal diseases. Twenty patients (aged 22 to 79 years) were divided into 2 groups, a group with meningeal disease comprising 9 patients with meningeal abnormalities (6, tumor dissemination; 3, infection) and a control group of 11 patients with unilateral IAC pathology whose healthy sides were included as controls. Six of the 9 patients in the group with meningeal disease showed bilateral enhancement; one showed unilateral enhancement. None of the control group showed enhancement in the healthy side. One patient with Ramsay Hunt syndrome showed only ipsilateral enhancement. Enhancement in the IAC fundus was frequently observed in patients with meningeal disease, even just 10 min after administration of contrast agent. This enhancement in the IAC fundus was never visible on T1-weighted 3D-FLASH images. PMID- 17139142 TI - Real-time functional MRI: development and emerging applications. AB - Real-time functional magnetic resonance imaging (fMRI) is an emerging technique for assessing the dynamic and robust changes in brain activation during an ongoing experiment. Real-time fMRI allows measurement of several processes within the brain as they occur. The extracted information can be used to monitor the quality of acquired data sets, serve as the basis for neurofeedback training, and manipulate scans for interactive paradigm designs. Although more work is needed, recent results have demonstrated a variety of potential applications for real time fMRI for research and clinical use. We discuss these developments and focus on methods enabling real-time analysis of fMRI data sets, novel research applications arising from these approaches, and potential use of real-time fMRI in clinical settings. PMID- 17139143 TI - A case of hepatocellular carcinoma treated by MR-guided focused ultrasound ablation with respiratory gating. AB - Focused ultrasound surgery (FUS) is a method of noninvasive focal thermal ablation. Temperature-sensitive phase-difference magnetic resonance (MR) imaging allows monitoring of the focal point and measurement of tissue temperature elevation in real time, ensuring delivery of a therapeutic dose. A newly developed respiratory monitoring system enables us to track liver tumors, which move with respiration. We report our initial experience using MR-guided FUS with respiratory gating in successfully treating a hepatocellular carcinoma 15 mm in diameter. PMID- 17139145 TI - [Humoral and cellular responses in innate immunity]. AB - The immune system is divided into innate and adaptive immunity. Either immunity consists of humoral and cellular responses, and immunity is maximized when both responses coordinately function. Adaptive immunity has been intensively studied, while it was only recently that we gained some understanding of innate immunity. In particular, cellular responses in innate immunity have been poorly understood compared with humoral responses. In addition, the mechanisms and roles of innate immune responses could be distinct between the organisms that possess both innate and adaptive immunity and those possessing only innate immunity. On the other hand, invading pathogenic microbes employ various strategies to inhibit the host immune system for their survival. I here summarize what needs to be known to gain a deeper understanding of the innate immune response. The readers are suggested to refer to the accompanying articles for more detailed description. PMID- 17139146 TI - [Intra- and extracellular recognition of pathogens and activation of innate immunity]. AB - One of the fundamental questions in innate immunity is how a large battery of invading pathogens is recognized by a limited number of germ line-encoding receptors. In Drosophila, peptidoglycan recognition protein (PGRP) family members have a crucial role in recognizing invading bacterial pathogens and in inducing immune reactions. PGRP-SA, -SD, and -SC1a are involved in recognizing gram positive bacteria and in activating the Toll pathway to produce antimicrobial peptides. PGRP-LC and -LE recognize diaminopimelic acid (DAP)-containing peptidoglycans, which are cell wall components of many gram-negative bacteria and some gram-positive bacteria, and activate the imd pathway to produce antibacterial peptides. In addition to the extracellular function of PGRP-LE to activate immune reactions in the hemolymph, PGRP-LE acts as an intracellular receptor for monomeric DAP-type peptidoglycans. Moreover, a version of PGRP-LE containing only the PGRP domain functions extracellularly as a CD14-like accessory factor, capable of enhancing PGRP-LC-mediated peptidoglycan recognition. Subsequent intracellular signaling is transduced through the RHIM like motif found in PGRP-LC and -LE. PMID- 17139147 TI - [The fruit fly as a tractable model animal for infectious diseases: implication for understanding of host-pathogen interaction]. AB - Use of invertebrate models of infection has given exciting insights into host pathogen interaction for a number of bacteria. In particular, this has revealed important factors of the host response with remarkable parallels in higher organisms. Recently, emerging of multi-drug resistant bacteria raises a requirement of developing new therapies such as controlling host defense system. Finding host factors that can purge bacteria from human body could give us a new concept of pharmaceutical targets. For this purpose, fruit flies, Drosophila melanogaster, has been used as a model animal for human infectious diseases and became a tractable tool for identifying novel gene products that can activate host defense mechanisms. In this review we will discuss about recent progress of Drosophila model of pathogen infection, which could imply a useful genetically tractable model for human infectious diseases. PMID- 17139148 TI - [Outer membrane remodeling of Salmonella typhimurium and host innate immunity]. AB - Pathogenic gram-negative bacteria, including Salmonella typhimurium, remodel their outer membrane to survive within host tissues and phagosomes. The remodeling includes modifications of lipid A, a membrane anchor portion of lipopolysaccharide. Lipid A modifications, such as palmitoylation, deacylation, addition of aminoarabinose, and addition of phosphoethanolamine, are beneficial for salmonellae to resist host innate immunity. Aminoarabinose attachment, phosphoethanolamine attachment, and palmitoylation of lipid A increase salmonellae resistance to cationic antimicrobial peptides. Lipid A deacylation and palmitoylation reduce its ability to activate the Toll-like receptor 4-MD-2 complex, suggesting that these modifications are beneficial for salmonellae to evade host innate immune recognition. These modifications are regulated transcriptionally by the two-component regulatory system PhoP-PhoQ, which is essential for S. typhimurium virulence. Lipid A modifications are also regulated posttranslationally. Aminoarabinose modification of lipid A represses deacylation of lipid A by PagL. The posttranslational regulation may be involved in S. typhimurium pathogenesis. PMID- 17139149 TI - [Bacterial strategies for escaping the bactericidal mechanisms by macrophage]. AB - Phagocytosis with macrophages provides a specialized mechanism for regulated ingestion and intracellular destruction of bacteria. Bacteria are first engulfed by endocytosis into a phagosome. The fusion of phagosomes and lysosomes releases toxic products that kill most bacteria and degrade them into fragments. Debris from dead bacteria is then released by exocytosis. However, some bacteria that survive within host phagocytes have evolved strategies to escape the bactericidal mechanisms associated with phagocytosis: i) antiphagocytosis (Yersinia), ii) escaping from the phagosome into cytoplasm (Listeria), and iii) remodeling their phagosome by inhibiting the maturation of phagosomes (Salmonella, Mycobacterium, Legionella). In this review, I first summarize various strategies by bacteria to avoid phagocytosis by emphasizing the steps that have been subverted by bacteria. Then, I highlight the mechanisms for surviving phagocytosis by Salmonella, with a focus on the induction of macrophage-apoptosis and modulation of membrane traffic in host cells. PMID- 17139150 TI - [Elimination of influenza virus-infected cells by phagocytosis]. AB - Infection with a variety of viruses induces apoptosis in host cells. This phenomenon may be considered to be a self-defense mechanism to avoid viral propagation. However, the growth of influenza virus is completed before host cells become dysfunctional due to apoptosis. To clarify the physiologic consequences of influenza virus-induced apoptosis, the fate of influenza virus infected cells was examined in vitro as well as in vivo. Influenza virus-infected cells were engulfed by macrophages in vitro, and virus propagation was almost completely inhibited. This phagocytosis was dependent on the specific recognition of the membrane phospholipid phosphatidylserine exposed on the surface of virus infected apoptotic cells by macrophages. In addition, the activity of viral neuraminidase expressed at the surface of virus-infected cells was necessary for the maximal level of phagocytosis. When mice infected with influenza virus were administered phagocytosis inhibitors, the level of lethality and inflammation in the lung were augmented. These results show that apoptosis makes influenza virus infected cells susceptible to phagocytosis by macrophages, and that this leads to a reduction in the extent of influenza pathology. PMID- 17139152 TI - [Chemical incidents and gathering information on toxicity]. AB - Major cases of chemical incidents and information on chemical agents and chemical terrorist attacks are outlined. Since the late 1990s, major incidents occurred consecutively, such as two cases of sarin attack in 1994 and 1995, an oil spill from a Russian oil tanker in the Japan Sea in 1997, arsenic poisoning in Wakayama in 1998, the criticality incident at Tokai-Mura in 1999 in Japan, and terrorist attacks on September 11, 2001, in New York. The importance of crisis management and cooperation among relevant organizations has been emphasized. To provide information for an appropriate and quick response in emergencies, we prepared a Web portal site for information on chemicals including chemical agents, a chemical incident database, and links to relevant Web sites. In intentional cases of poisoning caused by toxic chemicals in Japan, 111 cases were collected mainly from a newspaper database (1984-1999). Many copy-cat poisonings occurred, especially in 1984-1985 and in 1998 just after an arsenic poisoning incident in Wakayama. Many cases occurred in the laboratories of institutes, universities, and hospitals where various types of chemicals are used. PMID- 17139153 TI - [Simplified rapid test for poisons in emergency and critical care medicine]. AB - In emergency and critical care medicine, it is important to guess which poisons that patients have taken or been exposed to. The assumption and identification of save lives. Therefore an accurate screening system is required to treat acute poisoning patients in clinical toxicology. However, the ability of a medical center is not sufficient to analyze poisonous substances using analytical equipment. Moreover, the handling and maintenance of the equipment are tedious and costly. To improve these problems, a simple detection method should be established to identify poisons and to treat acute patients in emergency and critical care medicine. In our laboratory, various supports have been attempted for the training of analysts who cope with poisoning incidents and accidents due to toxic substances. Moreover, a simple detection method for toxic substances utilized in the medical center was developed without using expensive analysis apparatus. However, it is impossible to detect and identify chemical warfare agents in a clinical laboratory, because of possible secondary exposure to such dangerous substances in insufficient analytical laboratory equipment. Therefore it is necessary to contact related organizations possessing the proper facilities. PMID- 17139154 TI - [Analytical and on-site detection methods for chemical warfare agents]. AB - Chemical warfare agents (CWAs) are fast acting and sometimes lethal, even at low levels, and can be classified into nerve gases, blister agents, choking agents, blood agents, vomit agents, tear gases, and incapacitating agents. As countermeasures against CWA terrorism, detection and identification are important. In crisis management, monitoring of CWAs in public places and security checks at territorial borders, big event venues, and executive facilities are performed for protection against terrorism. In consequence management, on-site detection by first responders and laboratory analysis after on-site sampling and transfer are performed for minimization of terrorism damage, leading to personal protection, initial investigation, and emergency lifesaving. In incident management, laboratory analysis is performed to provide evidence at court trials for the prevention of future crimes. Laboratory analysis consists of pretreatment of on-site and casualty samples and instrumental analysis using GC-MS. However, CWAs are easily degraded, and thus are difficult to detect. Instead, it is useful to detect their metabolites and degradation products using tert butyldimethylsilyl derivatization GC-MS or direct LC-MS. Commercially available chemical detection equipment such as gas detection tubes and ion mobility spectrometers are used for on-site detection. We have evaluated the detection performance of such equipment and found that no equipment fulfills the required perfect performance of CWA detection sensitivity, accuracy, response time, return time, and operation. To overcome the drawbacks, we have adopted the monitoring tape method and counterflow introduction atmospheric pressure chemical ionization mass spectrometry and recommend the combination of commercial detection equipment and these new technologies for simultaneous, rapid detection of all CWAs. PMID- 17139155 TI - [Use of biosensors for detecting organophosphorus agents]. AB - The present paper reviews the use of electrochemical biosensors for detecting organophosphorus pesticides and nerve agents. Acetylcholine esterase (AChE) immobilized electrodes have been used for detecting AChE inhibitors including organophosphorus and carbamate pesticides. The sensors are composed of AChE and choline oxidase (ChOx) for converting the AChE-generated choline into betaine and hydrogen peroxide (H(2)O(2)), which is electrochemically oxidized at the electrode surface to produce the output signal of the sensor. In the presence of AChE inhibitors, the suppressed output signal of the sensor can be observed. If the sensors are operated in the presence of acetylthiocholine as a substrate of AChE, one can eliminate ChOx from the sensor design because enzymatically generated thiocholine is electrochemically active and thus directly oxidized at the electrode without using ChOx. Electron-transfer mediators such as tetracyanoquinodimethane have often been used for catalytically oxidizing thiocholine at the electrode set at less positive potential, which is effective in circumventing possible interference arising from oxidizing compounds in the sample solution. One of the drawbacks of the AChE-based biosensors in detecting organophosphorus pesticides and nerve agents arises from the fact that the sensors indirectly detect the signal based on the inhibition of the AChE catalyzed reaction. On the other hand, for directly obtaining the output signal, organophosphorus hydrolase (OPH) is immobilized on the electrode surface to prepare amperometric biosensors. OPH catalyzes the hydrolysis reaction of organophosphorus compounds to produce electrochemically active compounds such as p-nitrophenol and thiols from parathion and VX, respectively. Thus OPH-based sensors can be used for detecting these compounds directly. These biosensors would be useful for in-site measurements of organophosphorus pesticides and nerve agents because portable-type biosensors are easily fabricated at relatively low cost. PMID- 17139156 TI - [How I was enticed into molecular toxicology]. AB - This paper introduces one of our projects performed at Hokkaido University. During the course of pharmacokinetic studies of SM-12502, which was under development as an anti-platelet-activating factor agent, we found three individuals who showed a slow metabolic phenotype in its pharmacokinetics. Analyzing the genes for CYP2A6 from the three, we discovered that they had the whole CYP2A6 gene deletion (CYP2A6*4C). Genetically engineered Salmonella YG7108 cells expressing human P450 were established to compare the mutagen-producing capacity of the P450 enzymes for various N-nitrosamines. We found that CYP2A6 was involved in the metabolic activation of N-nitrosamines with relatively bulky alkyl chains such as a tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone (NNK), which has been known to cause lung tumors in rodents. Thus, to examine the hypothesis that individuals possessing the CYP2A6*4C have a reduced risk of cancer due to the lack of the metabolic activation of certain carcinogens in tobacco smoke, a case-control study was performed. The results clearly indicated a significant association between the CYP2A6 genotype and lung cancer risk in smokers. In contrast, there was no significant relationship between them in nonsmokers. In addition, our results showed that the reduced risk of cancer was caused by the reduced activity of CYP2A6. Thus it was expected that the inhibition of the enzyme would result in a reduced cancer risk caused by smoking. The results of experiments using mice which were treated with NNK, a carcinogenic nitrosamine contained in tobacco smoke, together with 8 methoxypsolaren, a strong inhibitor of CYP2A6, indicated that the inhibition of CYP2A6 completely abolished the occurrence of adenoma. PMID- 17139157 TI - [Development of efficient methods for synthesis of nitrogen-containing compounds using carbamates, acylhydrazines, and ammonia]. AB - For the efficient synthesis of divergent nitrogen-containing compounds of pharmaceutical and agricultural importance, the development of efficient, complementary, and new synthetic methodologies is essential. One of the key subjects is how to introduce nitrogen atoms in to organic molecules. This review summaries our recent efforts on this issue, focusing on the use of carbamates, acylhydrazines, and ammonia as nitrogen sources. PMID- 17139158 TI - [Sweet potato in pre-Columbian Polynesia--an overview]. AB - An overview is presented on the reports available so far on sweet potato, Ipomoea batatas, cultivated widely in Polynesia in the pre-Columbian era, with reference to possible ways and presumptive dates of transfer from the Americas to Polynesia, such as (1) Polynesian navigators' travel to Peru, (2) Peruvian fishermen's drift westward, (3) vessel drift, (4) seed drift, (5) root-tuber drift, and (6) transport by birds. The author supports the case (1) as most plausible. Ganshu or Ganchu described in the old Chinese herbal books is identified as Dioscorea esculenta. An introduction of the tuber to China and Japan is briefly mentioned. PMID- 17139159 TI - Usefulness of a nutrition assessment system for parenteral/enteral nutrition therapy. AB - Since a nutrition support team (NST) began to work in our hospital in March, 2003, we constructed our original nutrition assessment system that supports the prescription formulation of total parenteral nutrition (TPN). However, in daily NST activities, the re-evaluation of this system became necessary because of a high incidence of enteral nutrition (EN) and marked revisions in the dietary reference intakes in Japanese (7th revision). Therefore, we improved this system and added a prescription formulation support function that is also applicable to EN, and also added a function that automatically calculates the necessary doses of nutrients that tend to become deficient in patients with decubituses. This new system allowed the selection/evaluation of EN solutions in a short time with consideration of the 7th revision, and readily identified deficient nutrients and their levels in decubitus patients. We used this system in patients with high level malnutrition complicated by decubituses and observed certain treatment effects. PMID- 17139160 TI - Quantification and 24-hour monitoring of mycophenolic acid by high-performance liquid chromatography in Japanese renal transplant recipients. AB - We developed a rapid, simple, and sensitive high-performance liquid chromatography method with UV detection for the quantitative determination of mycophenolic acid (MPA) in human plasma. MPA and the internal standard (naproxen) were separated using a mobile phase of 0.04 M H(3)PO(4)-acetonitrile-methanol (3:3:4 v/v/v) over a CAPCELL PAK C18 MG column. A flow-rate of 0.5 ml/min was used at ambient temperature and sample detection was carried out at 254 nm. The assay required only 100 microl of plasma and involved liquid-liquid extraction, which gave high recovery (>94%). The lower limit of quantification for MPA was 0.05 microg/ml. Inter- and intra-day coefficients of variation were less than 9.6% and accuracies were within 9.3%. Additionally, we validated this method in 24-hour monitoring of plasma MPA concentrations after mycophenolate mofetil (MMF) morning and evening administration in 40 Japanese renal transplant recipients with 1.5 g/day MMF. The time to reach the maximum (11.7 microg/ml) and second peak (4.5 microg/ml) of MPA after morning 0.75 g MMF administration was 2.6 h and 9.0 h, and time to reach maximum (10.5 microg/ml) after evening 0.75 g administration was 4.0 h. PMID- 17139161 TI - Evaluation of the state of active ingredients in pharmaceutical preparations using fourier transform-Raman difference spectra. AB - To examine the pharmaceutical application of Fourier transform (FT)-Raman spectroscopy, the state of active pharmaceutical ingredients (APIs) in a preparation of several forms was evaluated by investigating the Raman difference spectra between the preparation and excipient. The difference spectra indicated that APIs in alacepril tablets, caffeine sustained-release granules, and quinidine sulfate granules remained unchanged after the manufacturing process. However, the state of sparfloxacin in nanoparticles changed, although it remained unchanged in tablets or powders. These results show that the FT-Raman difference spectrum is expected to be utilized in the field of quality control of crystalline pharmaceutical preparations. PMID- 17139162 TI - Effect of formulation factors on in-vitro permeation of diclofenac from experimental and marketed aqueous eye drops through excised goat cornea. AB - The effect of formulation factors on permeation of diclofenac from some experimental and marketed aqueous eye drops through excised goat cornea was evaluated. Raising the pH of formulation from 6.0 to 8.0 or diclofenac concentration from 0.05 to 0.15% (w/v) or adjusting tonicity with mannitol or addition of viscolizing agent decreased apparent permeability coefficient (Papp). Formulation (pH 7.4) containing sodium metabisulfite or EDTA or combination of methyl and propyl paraben showed significantly (p<0.05) higher Papp whereas benzalkonium chloride (BAC) had no effect and sorbic acid (SA) had reduced permeation. Surprisingly marketed drops containing BAC or SA, showed significantly (p<0.05) higher Papp and decreased in the order of Difen>Voveran>NSAID>Dicol>Diclolab. Lower pH (7.1-7.3) and surface tension of drops indicating presence of surfactant, could mediate increased permeation and presence of buffer could cause irritation on in vivo instillation. The marketed formulations showed corneal hydration >83% suggesting corneal damaging potential. PMID- 17139163 TI - Effects of benidipine in a rat model for experimental angina. AB - To compare the antianginal effects of 1,4-dihydropyridine-type calcium-channel blockers, we evaluated the effects of benidipine, amlodipine, nifedipine, and efonidipine on vasopressin-induced myocardial ischemia in rats, an experimental model of angina. Intravenous administration of benidipine (3 microg/kg), amlodipine (1000 microg/kg), and nifedipine (100 microg/kg) suppressed the vasopressin-induced S-wave depression, an index of myocardial ischemia. Efonidipine (100 microg/kg, i.v.) tended to inhibit the S-wave depression. At the antianginal dose of each drug, amlodipine, nifedipine, and efonidipine decreased blood pressure significantly, whereas benidipine had little effect on blood pressure at a dose of 3 microg/kg. These results indicate that benidipine, unlike the other 1,4-dihydropyridine-type calcium-channel blockers examined in this study, inhibits vasopressin-induced coronary vasospasm with fewer undesirable effects such as hypotension in rats, suggesting that benidipine may be useful in the treatment of angina pectoris. PMID- 17139164 TI - Effect of shikonin on human breast cancer cells proliferation and apoptosis in vitro. AB - Shikonin, isolated from the plant Lithospermum erythrorhizon Sieb. Et Zucc, has been reported to induce apoptosis in several tumor cells. However, such effect of shikonin on human breast cancer cells has not been reported. Thus, in the present study, whether shikonin could induce MCF-7 human breast cancer cell apoptosis was investigated. The results showed that shikonin (2.5-80 microM) induced MCF-7 cell death in a time- and dose-dependent manner, as measured by MTT assay. The IC(50) of a 24 h, 48 h and 72 h time course for MCF-7 cells was 7.4+/-0.4, 6.3+/-0.6 and 3.9+/-0.5 microM, respectively. Cellular morphology observation showed that MCF-7 cells underwent marked apoptotic morphological changes upon treatment with 10 microM shikonin compared with the untreated control. Flow cytometric analysis of shikonin-treated MCF-7 cells showed that the ratio of the apoptotic DNA fragmentation increased in a dose-dependent manner. The present study demonstrated for the first time that the cytotoxic effect of shikonin on MCF-7 cells underwent apoptosis process. PMID- 17139165 TI - A systematic review of the psychological risk factors associated with near fatal asthma or fatal asthma. AB - Psychological factors such as anxiety, depressive disorders and/or personality disorders may predispose patients with asthma to near fatal asthma (NFA) or fatal asthma (FA). NFA is defined by an asthma exacerbation resulting in respiratory arrest requiring mechanical ventilation or a pCO(2) >or=45 mm Hg. Most studies have used the case-control study design. Several studies analyzing the effects of psychological factors on the risk of NFA or FA have shown conflicting results. We reviewed all of the literature found by the systematic search done of psychological factors on the risk NFA or FA. A MEDLINE search identified 423 articles between 1960 and March 2006. Seven case-controlled studies were identified following strict applications of the inclusion and exclusion criteria. Due to the significant heterogeneity in the measurement of the psychological factors, a summary statistic was not calculated. The trial characteristics were tabulated and qualitative trends were observed to explain the heterogeneity in the results of the studies. Recommendations on future studies in the field are outlined in detail. Following a systematic assessment of all published studies, we cannot conclude that psychological factors increase the risk of NFA and FA. PMID- 17139166 TI - Helsingborg Declaration 2006 on European stroke strategies. AB - The Second Consensus Conference on Stroke Management took place from 22 to 24 March 2006 in Helsingborg, Sweden. The meeting was arranged by the International Stroke Society, endorsed by the European Stroke Council and International Stroke Society, and co-sponsored by the WHO Regional Office for Europe. It was arranged in collaboration with the European Region of the World Confederation for Physical Therapy and the European Association of Neuroscience Nurses. The patients' organization Stroke Alliance for Europe also participated. The meeting adopted the Helsingborg Declaration 2006 on European Stroke Strategies, a statement of the overall aims and goals of five aspects of stroke management (organization of stroke services, management of acute stroke, prevention, rehabilitation, evaluation of stroke outcome and quality assessment) to be achieved by 2015. PMID- 17139167 TI - An inverse correlation between Interleukin-6 and select gene promoter methylation in patients with gastric cancer. AB - BACKGROUND: Both serum IL-6 levels and CpG island methylation have been shown to have prognostic significance in gastric cancer, it was suggested that an important link existed between IL-6 and methylation of cancers. AIM: To investigate the prognostic value of IL-6 serum level and the association between serum IL-6 levels and CpG island methylation at p16, DAPK, MGMT and E-cadherin in patients with gastric cancer. PATIENTS AND METHODS: Methylation status was assessed by MSP in 75 surgical specimens of gastric adenocarcinoma. IL-6 serum levels were measured by chemiluminescent enzyme immunoassay (CLEIA). RESULTS: Methylation of p16, DAPK, MGMT, and E-cadherin were present in 53, 48, 32, and 59% of patients. Patients with tumors methylated at p16 and DAPK had lower serum levels of IL-6 compared to unmethylated tumors (1.8 vs. 4.8 pg/ml, p = 0.01 for p16; 1.5 vs. 6.2 pg/ml, p = 0.0001 for DAPK). But there was no difference with MGMT and E-cadherin methylation status. Serum IL-6 levels were also associated with TNM stage (p = 0.001), depth of tumor invasion (p = 0.002), lymphatic invasion (p = 0.01), vascular invasion (p = 0.008), metastasis (p = 0.002) and signet cell histology (p = 0.001). CONCLUSION: IL-6 is of prognostic value for patients of gastric cancer. Low serum IL-6 levels were associated with p16 or DAPK gene methylation in patients with gastric cancer. PMID- 17139168 TI - Emergent fetal intracardiac transfusion for thrombocytopenia and acute hypovolemia due to cordocentesis-associated hemorrhage in parvovirus-induced hydrops. AB - OBJECTIVES: To demonstrate the utility of fetal intracardiac transfusion to correct acute fetal hypovolemia and thrombocytopenia in fetal Parvovirus infection. METHODS: Intracardiac transfusion in a 19-week gestation was indicated due to cordocentesis-associated hemorrhage. RESULTS: Intracardiac transfusion resulted in correction of acute bradycardia, anemia and thrombocytopenia and persistent umbilical cord hemorrhage following attempted intravascular transfusion. CONCLUSIONS: This case illustrates the importance of anticipating both thrombocytopenia and anemia in fetal Parvovirus infection and how an intracardiac approach can be employed in the setting of acute, life-threatening hemorrhage. PMID- 17139169 TI - Advanced combined abdominal and intrauterine pregnancy: a case report. AB - We present a case of simultaneous abdominal and intrauterine pregnancy following in vitro fertilization and embryo transfer diagnosed during the second trimester. The patient had a recent history of hysteroscopic metroplasty due to in utero exposition to diethylstilboestrol. The pregnancy was managed conservatively with a favorable outcome for the mother and both fetuses. PMID- 17139170 TI - A study to determine if human umbilical cord hematopoietic stem cells can survive in baboon extra-embryonic celomic fluid: a prerequisite for determining the feasibility of in-utero stem cell xeno-transplantation via celocentesis. AB - OBJECTIVES: To determine if: (1) human umbilical cord stem cells could survive for 20 h in extra-embryonic celomic fluid obtained at 40 days of development from baboon pregnancies by ultrasound-guided celocentesis and, (2) human hematopoietic stem cell survival could be enhanced by adding increasing concentrations of hematopoietic stem cell culture medium to the celomic fluid. METHODS: CD34+ cells were isolated from umbilical cord blood using a magnetic activated cell sorter and flow cytometry. Cells were then cultured in five platforms containing different combinations of baboon extra-embryonic celomic fluid and hematopoietic stem cell culture medium to determine cell survival kinetics over a 20-hour period in each of the conditions. RESULTS: Human umbilical cord stem cells can survive for at least 20 h in baboon's celomic fluid. Chi-square for linear trends demonstrated that the number of viable cells was significantly enhanced by adding the increasing concentrations of culture medium to the celomic fluid (29% cell survival in pure celomic fluid, 48% at 1:8, 50% at 1:2, 54.5% at 1:1, 61% in pure culture medium; p < 0.001). CONCLUSIONS: Human umbilical cord stem cells survive in baboon celomic fluid and cell survival improves when the celomic fluid is mixed with greater concentrations of hematopoietic stem cell culture medium. Based on these findings, future in-vivo experiments in the pregnant baboon animal model can be directed at determining whether in-utero injection of human hematopoietic stem cells prior to 10 weeks of pregnancy can lead to permanent chimeras. PMID- 17139171 TI - Unusual manifestation of acute hepatic porphyria in pregnancy. AB - A 22-year-old para I/gravida II developed psychiatric symptoms at 8 weeks of gestation. Subsequently neurological symptoms with seizures developed leading to a status epilepticus with continuing seizures at week 14. Anticonvulsive therapy had little effect in alleviating the seizures and the condition of the patient rapidly deteriorated. A sudden reddening of her urine lead to the diagnosis of acute hepatic porphyria confirmed by laboratory tests. After extensive discussion with the patient's family it was decided to terminate the pregnancy at week 16. Within hours after pregnancy termination the seizures stopped and the patient recovered without any neurological deficits. Acute hepatic porphyria can be triggered by pregnancy and usually presents with gastrointestinal symptoms and personality changes. In its rare neurological manifestation it can lead to untreatable convulsions which leave no option but to terminate the pregnancy. PMID- 17139172 TI - Emergency peripartum hysterectomy in a tertiary obstetric center: 8-year evaluation. AB - OBJECTIVE: The aim of the study was to present the incidence, indications, and operative morbidity and mortality in pregnant women undergoing emergency peripartum hysterectomy (EPH) at a tertiary obstetric institution. METHODS: In this retrospective clinical study, performed during the period 1995-2003, 17 EPH procedures were recorded in a total of 21,659 deliveries carried out at Department of Gynecology and Obstetrics, Osijek Clinical Hospital in Osijek, Croatia. Data on the incidence of EPH in total number of deliveries, rate of EPH in vaginal delivery and cesarean section, indications for EPH, and maternal and fetal/early neonatal morbidity and mortality were derived from operative protocols and medical records of hospitalized patients. RESULTS: During the 8 year study period, the incidence of EPH in total number of deliveries was 0.078%. Out of 17 EPH procedures, 5 (29.41%) were performed after vaginal delivery and 12 (70.59%) during cesarean section, elective in five and urgent in seven cases. The indications for EPH included severe peripartum hemorrhage with placenta previa in four, placenta previa percreta in four, various forms of invasive malplacentation (placenta accreta, increta, percreta) in five, uterine rupture in two cases, and atony along with massive retroperitoneal hematoma due to rupture of periuterine vascular bundle during cesarean section in one multipara. EPH was carried out in 12 multipara and five primipara. Lesions of urinary bladder occurred in three cases and were managed by suture. Twelve patients received blood transfusion, whereas development of hemorrhagic shock necessitated transfer to Intensive Care Unit in three patients. No late complications or maternal mortality were recorded. Sixteen total hysterectomies and one supracervical hysterectomy were performed. One case of intrauterine fetal death was caused by total abruptio placenta and uterine rupture during the patient's transport from a primary obstetric institution. CONCLUSION: Invasive malplacentation is a major isolated risk factor for EPH, as shown in the present study. Other risk factors for EPH are massive hemorrhage because placenta previa, uterine atony and uterine rupture associated with multiparity, and previous cesarean section. A great proportion of EPH procedures can be prevented by the introduction of compressive operative methods such as B-Lynch suture in the obstetric algorithms, which will certainly favorably reflect in future fertility and genital health of the female population. PMID- 17139173 TI - T2 quantifications of lungs in the fetal lamb with experimentally-induced congenital diaphragmatic hernia. AB - OBJECTIVE: As an aid to evaluate lung hypoplasia, we investigated the difference between T2 value in fetal lungs of lambs with surgically-induced left-sided diaphragmatic hernia (DH) and gestational age (GA)-matched control littermates (normal [NL]). METHODS: Lungs were divided into two groups: DH (n = 4) and NL (n = 6). DH was induced at 65-75 days GA (term = 145 days). Fetal MRI was performed (mean GA: 120 days) with half-Fourier acquisition single-shot turbo spin-echo (TE: 60 ms) and rapid acquisition with relaxation enhancement (TE: 350 ms) in the same location (1.5-T Philips, Gyroscan, Best, The Netherlands). T2 of each lung was calculated for multiple regions of interest by taking natural logarithm of signal-to-noise ratio. Mean T2 was compared between DH and NL (unpaired analysis for entire group). Paired comparison between left/right lung was made within DH and NL. RESULTS: Unpaired analysis showed significantly lower T2 of left respectively right lungs in DH (p = 0.02 [respectively] 0.05]) compared to NL (n = 6), as well as between the T2 of all DH versus NL (p = 0.001). In DH, calculated T2 appeared to be lower in left than in right lungs (difference ranged from -2 to +49%). In NL, left and right lungs showed comparable T2. CONCLUSION: Measurement of T2 signal intensity in DH lungs is feasible and show lower T2 in comparison to NL lungs. Left lungs from lambs with DH show lower T2 than right lungs. PMID- 17139174 TI - Ultrasound assessment prior to laser photocoagulation for twin-twin transfusion syndrome for predicting intrauterine fetal demise after surgery in Japanese patients. AB - OBJECTIVE: To evaluate the use of ultrasound before selective laser photocoagulation of communicating vessels (SLPCV) for twin-twin transfusion syndrome in predicting intrauterine fetal demise (IUFD). METHODS: Fifty-five patients underwent SLPCV in Japan. Fetal biometry and Doppler studies of the umbilical artery, ductus venosus, and umbilical vein were performed prior to SLPCV. The visualization of the bladder and hydrops was recorded. Association between the parameters and IUFD was analyzed using multiple logistic regression analysis. The study was approved by the Institutional Review Board and patients gave their informed consent. RESULTS: The IUFD incidence was 25.5% (14/55) in the donors and 12.7% (7/55) in the recipients. Twelve donors and 4 recipients of them ended in unexplainable IUFD. In the analysis of 53 donors, absent or reversed end diastolic flow of umbilical artery (UAAREDF) was only associated with IUFD (p = 0.016). No parameters could predict IUFD in 52 recipients. CONCLUSIONS: UAAREDF may be useful for predicting IUFD of the donor after SLPCV. PMID- 17139175 TI - Prenatal diagnosis and genetic counseling in a case of spina bifida in a family with Waardenburg syndrome type I. AB - OBJECTIVE: Waardenburg syndrome type I (WS I) is an autosomal dominant inherited disorder with an incidence of 1:45,000 in Europe. Mutations within the PAX3 gene are responsible for the clinical phenotype ranging from mild facial features to severe malformations detectable in prenatal diagnosis. METHODS: Here, we report a four-generation family with several affected members showing various symptoms of WS I. We diagnosed the syndrome first in a pregnant young woman; she was referred because of a spina bifida in prenatal diagnosis. We performed clinical genetic investigations and molecular genetic analysis in all available family members. RESULTS: The phenotype displays a wide intra-familial clinical variability of pigmentary disturbances, facial anomalies and developmental defects. Molecular studies identified a novel splice site mutation within the PAX3 gene in intron 5 in all affected family members, but in none of the unaffected relatives. CONCLUSIONS: This case demonstrates the prenatal diagnosis of spina bifida in a fetus which leads to the initial diagnosis of WS I. Further studies could identify a private splice site mutation within the PAX3 gene responsible for the phenotype in this family. PMID- 17139176 TI - Re: Umbilical cord diameter at 11-14 weeks of gestation: relationship to nuchal translucency, ductus venous blood flow and chromosomal defects. PMID- 17139178 TI - Fatal bleeding following delivery: a manifestation of the vascular type of Ehlers Danlos' syndrome. AB - INTRODUCTION: The vascular form of Ehlers-Danlos' syndrome (type IV) is a potentially lethal genetic condition because of rupture of major arteries, often in the peri-partum period. CASE REPORT: We report a 31-year-old primipara who died from a rupture of the right subclavian artery. The patient had several symptoms and signs typical of the disease. The rupture occurred during the expulsion-phase of delivery but was recognized only on day 9. CONCLUSION: Early recognition is crucial to avoid maternal mortality due to this genetic disorder. Once the condition is suspected, the clinical diagnosis is straightforward. PMID- 17139179 TI - Risk indicators of pre-eclampsia in north Jordan: is dental caries involved? AB - BACKGROUND: Despite a huge body of published research, understanding the basic pathology that leads to pre-eclampsia is still incomplete. This study was conducted among women in north Jordan to determine factors associated with pre eclampsia and to determine if dental caries was involved. METHODS: A case-control study was carried out among women who delivered at Princess Badea Teaching Hospital, Irbid, North Jordan, between September 1, 2004 and January 1, 2005. 115 women with pre-eclampsia and 230 normotensive controls were included in the study. Information regarding participants' demographics, antenatal history and family history were collected through personal interviews. Several oral parameters were measured for each participant. RESULTS: In multivariate analysis, factors found to be associated with increased odds of pre-eclampsia were age > or =35 years (adjusted OR 2.20; 95% CI 1.08, 4.48), nulliparity (adjusted OR 2.73; 95% CI 1.38, 5.39), emotional stress during pregnancy (adjusted OR 4.22; 95% CI 1.79, 9.90), maternal history of pre-eclampsia (adjusted OR 16.04; 95% CI 5.82, 44.22), family history of cardiovascular diseases (adjusted OR 2.82; 95% CI 1.22, 6.51), family history of pre-eclampsia (adjusted OR 23.26; 95% CI 5.07, 106.78), twin births (adjusted OR 37.79; 95% CI 4.22, 338.51), and Body Mass Index (BMI). In comparison with women with pre-pregnancy BMI <25, adjusted OR were 1.97, 95% CI 1.02, 3.81 for women with BMI from 25 to 29.9, and 3.31, 95% CI 1.36, 8.03 for women with BMI > or =30. Mean decayed teeth surface was found to be associated with increased odds of pre-eclampsia (adjusted OR 1.14, 95% CI 1.03, 1.27). CONCLUSION: This study demonstrated no difference to reported risk indicators of pre-eclampsia in other populations, but adds that dental caries may have an association with increased odds of pre-eclampsia. PMID- 17139180 TI - Hypoxia represses the differentiation of Rcho-1 rat trophoblast giant cells. AB - BACKGROUND: A hypoxic environment is known to be essential for early placentation. A low oxygen tension induces hypoxia-inducible factor-1 (HIF 1alpha) which may play an important role as a transcription factor in maintaining the proliferative and undifferentiated phenotype in human trophoblasts. METHODS: We analyzed the effect of a low oxygen tension on the rat trophoblast giant cell differentiation pathway using Rcho-1 cells which were derived from rat choriocarcinomas and consist of trophoblast stem cells. RESULTS: We found that a low oxygen tension suppressed the morphological changes and steroidogenesis during differentiation. The anticipated downregulation of the Id-1 transcription factor, a negative regulator of trophoblast giant cell differentiation, was not observed in the hypoxic environment. On the other hand, deferoxamine, which mimics hypoxia and induces HIF-1alpha, caused downregulation of Id-1 transcription factor and trophoblast giant cell differentiation. CONCLUSION: These results indicate that hypoxia represses rat trophoblast giant cell differentiation via an HIF-1alpha-independent pathway. PMID- 17139181 TI - Inhibition of SARS-CoV gene expression by adenovirus-delivered small hairpin RNA. AB - OBJECTIVE: Severe acute respiratory syndrome (SARS) is a highly contagious and lethal disease caused by a new type of coronavirus, SARS-associated coronavirus (SARS-CoV). Currently, there is no efficient treatment and prevention for this disease. We constructed recombinant adenoviral vectors that can express shRNAs, which inhibited the expression of SARS-CoV genes effectively in mammalian cells. METHODS: In this study, we designed several plasmids that express small hairpin RNA molecules (shRNA) specifically targeting to the genes encoding for the SARS CoV nucleocapsid (N) protein and envelope (E) protein, respectively. Effective shRNA molecules to the viral genes were screened and identified, and then constructed into adenovirus vectors. The effects of adenovirus-delivered small hairpin RNA on SARS-CoV gene expression were determined by RT-PCR, Western blot, and luciferase activity assays. RESULTS: The levels of viral mRNAs and viral proteins of the targets were significantly decreased or completely inhibited in cell lines after being infected with the recombinant adenoviruses that expressed specific shRNA molecules. CONCLUSIONS: Since many cell types can be efficiently infected by adenovirus, recombinant adenoviruses could serve as an alternative powerful tool for shRNA delivery and for gene suppression, especially when the targeted cells are resistant to transfection by DNA or RNA. With availability of high titers of adenoviruses and uniform and rapid infection, this approach would have foreseeable wide applications both in experimental biology and molecular medicine. PMID- 17139182 TI - Human coronavirus infection among children with acute lower respiratory tract infection in Thailand. AB - OBJECTIVE: This study was performed to further identify the previously uncharacterized human coronavirus 229E (hCoV-229E) and human coronavirus OC43 (hCoV-OC43) in Thailand by using the RT-PCR technique. In addition, we performed this study in order to delineate the prevalence, the potential clinical impacts and evaluation of the genetic characterization of this pathogen in young children who presented with acute lower respiratory tract infections (ALRI). METHODS: We obtained nasopharyngeal secretions (NPs) from 226 children <5 years of age who were either attending the outpatient department or hospitalized with ALRI from March 2002 to July 2003. All clinical, laboratory, RT-PCR, direct sequencing and phylogenetic analysis data were collected and analyzed. RESULTS: Of the 226 NPs samples from infants and young children presented with ALRI, 8 (3.54%) were positive for hCoV-229E, 2 (0.88%) were positive for hCoV-OC43, and 1 (0.44%) had co-infection. The following clinical presentations were noted: fever (100%), rhinitis (44%), acute bronchiolitis (44%), viral pneumonia (33%), viral pneumonia triggering asthma exacerbation (11%) as well as viral pneumonia causing BPD exacerbation (11%). All positive samples were subjected to direct sequencing. The amino acid sequences had 82-99% similarity to previous sequences stored in the GenBank database. CONCLUSION: The molecular technique we applied to detect human coronavirus appears justified as a valuable diagnostic approach to elucidate the prevalence, cause and clinical implications of ALRI among infants and young children. PMID- 17139183 TI - Intra- and intergenotypic variations among human cytomegalovirus gB genotypes. AB - OBJECTIVES: To study genotypic variations among human cytomegalovirus (HCMV) gB genotypes in clinical samples of Thai patients. METHODS: gB genotyping of 31 HCMV DNA-positive clinical samples were determined by PCR-RFLP, gene cloning and DNA sequencing methods. RESULTS: Eight gB1, 7 gB2, 7 gB3, 6 gB untype (UT1 and UT2) and 3 mixed gB genotypes were first identified by PCR-RFLP. All 3 mixed gB genotype samples and 1 gB2 sample were cloned and confirmed gB genotype by PCR RFLP. Altogether, 57 strains (27 unique types and 30 transformants) were further analyzed by DNA sequencing method. Discordant results between PCR-RFLP and DNA sequencing methods were demonstrated in 5 samples and 1 clone. The gB UT1 and UT2 were all classified as gB1 except 1 sample of UT1 (9D), suggesting a new variant. Each genotype had a similarity of more than 97%, whereas strains of different genotypes had 77.71-92.75% homology. The most divergent type was gB3. CONCLUSIONS: Intra- and intergenotypic variations among strains were demonstrated either in individual or distinct patients. Intragenotypic variation in a person occurs possibly due mainly to a point mutation mechanism rather than reinfection of a new gB genotype. PMID- 17139184 TI - Entry into and production of the Japanese encephalitis virus from C6/36 cells. AB - OBJECTIVES: The mosquito-borne Japanese encephalitis virus is a leading cause of encephalitis worldwide. However, few studies have investigated the kinetics of Japanese encephalitis virus internalization and production in mosquito cells, and fewer still have investigated the nature of the molecules involved in the binding of the virus to mosquito cells. METHODS: Using the Aedes albopictus/Stegomyia albopicta-derived C6/36 cell line, Japanese encephalitis virus internalization and production were assayed by standard plaque assay, and virus binding was investigated through preinfection enzymatic treatment of cells and virus overlay protein-binding assay of membrane fractions in native and denaturing gels. RESULTS: The internalization of the virus was nonlinear, and intracellular infective viruses were detected 8 h after infection and exported to the medium 10 h after infection. The internalization of the virus was primarily mediated by protein elements, and several bands were observed after overlay assay to membrane proteins, although mass spectroscopy was unable to identify candidate proteins. Soluble laminin produced a marginal, but dose-dependent inhibition of infection. CONCLUSIONS: These results suggest that the mechanism of Japanese encephalitis entry, production, attachment and receptor usage are distinct from those employed by the dengue viruses. PMID- 17139185 TI - Immunologic analysis induced by DNA vaccine encoding E protein of Beijing-1 strain derived from Japanese encephalitis virus. AB - OBJECTIVE: We have compared the gene expression and DNA immunization efficacy encoding prME and E proteins of a different strain (JaGAr-01) derived from Japanese encephalitis virus. This study aimed to construct a recombinant encoding E protein of the Beijing-1 strain derived from Japanese encephalitis virus and analyze the humoral, cellular and protective immunity induced by the above recombinant. METHODS: The recombinant pJBE containing E (1,500 bps) gene from the Beijing-1 strain of Japanese encephalitis virus was constructed and then transfected into the HepG2 cell line by liposome fusion. The expression of E (about 53 kD) protein in transfected cells was analyzed by Western blot using a specific anti-JEV-E antibody. BALB/c mice were vaccinated with 3 microg of pJBE by the gene-gun technique. JaGAr-01 and Beijing-1 strains (10(5) PFU/100 microl) of Japanese encephalitis virus were given to BALB/c mice by intraperitoneal injection 3 weeks after double DNA immunization with a lethal virus challenge. BALB/c mice were observed for 21 days after challenge. An 80% plaque reduction neutralization test was performed to titrate the neutralization antibody before and after viral challenge. A lactate dehydrogenase activity release test was used to examine cytotoxic T lymphocyte activity after double DNA immunization. RESULTS: The expression of about 53 kD protein associated with pJBE was determined in transfected HepG2 cells with specific anti-JEV-E antibody. A higher level of neutralization antibodies and the cytotoxicity effect were induced with pJBE immunization using the gene-gun technique, and were similar to those induced with inactivated vaccine derive from the Beijing-1 strain of Japanese encephalitis virus. Balb/c mice immunized with pJBE survived the challenge with the different strains of Japanese encephalitis virus; however, Balb/c mice immunized with inactivated vaccine did not survive the challenge with the JaGAr 01 strain of Japanese encephalitis virus at all. CONCLUSIONS: DNA vaccine containing the E protein gene derived from Japanese encephalitis virus can provide not only better efficacy including humoral and cellular immunity, but also cross-protection against infection with homologous and heterologous Japanese encephalitis virus. PMID- 17139186 TI - Effects of mitogen-activated protein kinase inhibitors on tumor necrosis factor alpha gene expression and apoptosis induction in cultured human fetal membrane chorion cells infected with influenza virus. AB - OBJECTIVES: We investigated the involvement of p38 mitogen-activated protein (MAP) kinase in tumor necrosis factor (TNF)-alpha gene expression, apoptosis induction and virus replication in cultured human fetal membrane chorion cells infected with influenza virus. METHODS: Influenza virus-infected chorion cells were incubated in the absence or presence of inhibitors of p38 MAP kinase, SB203580 and SB202190. TNF-alpha mRNA and hemagglutinin viral RNA (HA vRNA) were amplified with reverse transcriptase-polymerase chain reaction techniques. TNF alpha protein concentrations were determined by enzyme-liked immunosorbent assay. The extent of apoptosis induction was estimated by DNA agarose gel electrophoresis. Pyrrolidine dithiocarbamate (PDTC) and ribavirin, which have been shown to inhibit apoptosis induction via the inhibition of viral gene replication, were used as positive control reagents. RESULTS: PDTC and ribavirin inhibited the accumulation of TNF-alpha mRNA and HA vRNA in the virus-infected chorion cells, resulting in the suppression of TNF-alpha protein secretion. Both SB203580 and SB202190 suppressed TNF-alpha protein secretion, but not the accumulation of TNF-alpha mRNA as well as HA vRNA and the induction of apoptosis. CONCLUSIONS: These results suggest that p38 MAP kinase pathway is critical in TNF alpha gene expression at a post-transcriptional level but not in the apoptosis induction and influenza virus replication in cultured chorion cells. PMID- 17139187 TI - Vasopeptidase inhibition normalizes blood pressure and restores endothelial function in renovascular hypertension. AB - BACKGROUND/AIMS: Vasopeptidase inhibitors by definition inhibit both angiotensin converting enzyme (ACE) and neutral endopeptidase (NEP), therefore they may exceed the effect of ACE inhibitors in the treatment of hypertension. The present study investigated the effect of the vasopeptidase inhibitor AVE7688 in comparison to the ACE inhibitor ramipril on systolic blood pressure (SBP) and endothelial function in renovascular hypertension. METHODS: Wistar-Kyoto rats with renovascular hypertension (two-kidney one-clamp-model) were randomized 2 weeks after unilateral clamping of the right renal artery for 3 weeks' oral treatment with either AVE7688 (30 mg/kg/day), ramipril (1 mg/kg/day) or placebo. SBP was measured by the tail-cuff method and endothelium-dependent and independent vascular function was assessed in isolated preconstricted (norepinephrine 10(-7) mol/l) aortic rings as relaxation to acetylcholine (10( 10)-10(-4) mol/l) and sodium nitroprusside (10(-10)-10(-4) mol/l), respectively. RESULTS: Two weeks after clamping, SBP was significantly elevated (196 +/- 16 vs. 145 +/- 8 mm Hg for sham-operated rats; p < 0.01) and further increased in placebo-treated animals to 208 +/- 19 mm Hg. Treatment with AVE7688 and ramipril had a similar blood pressure-lowering effect (119 +/- 8 and 124 +/- 10 mm Hg, respectively; p < 0.01 vs. placebo). Maximum endothelium-dependent relaxation was reduced in hypertensive rats (72 +/- 6 vs. 99 +/- 7% in control rats; p < 0.05). Endothelium-dependent relaxation was restored by AVE7688 (101 +/- 6%) and ramipril (94 +/- 8%), respectively, whereas endothelium-independent relaxation was comparable in all groups. CONCLUSION: In renovascular hypertension the vasopeptidase inhibitor AVE7688 exhibited similar blood pressure-lowering and endothelial protective properties as compared to the ACE inhibitor ramipril. Therefore, in high renin models of hypertension, vasopeptidase inhibition may be considered an alternative treatment option to ACE inhibition. PMID- 17139188 TI - Could streptococcal erythrogenic toxin B induce inflammation prior to the development of immune complex deposits in poststreptococcal glomerulonephritis? AB - Acute poststreptococcal glomerulonephritis (APSGN) is a consequence of the immune response to streptococcal antigens with further in situ antigen-antibody interaction and deposition of circulating immune complexes, resulting in the activation of complement and the inflammatory process. These events are related to a previous antibody response. However, early renal events, when circulating streptococcal antigens bind to the kidney during streptococcal infection, remain unknown. Cationic streptococcal erythrogenic toxin type B (ETB) and its precursor (ETBP) are largely produced by nephritogenic streptococci and have high affinity for anionic glomerular structures. Renal deposition of ETB/ETBP makes conceivable a possible interaction between these streptococcal proteins with intrinsic glomerular cells or infiltrating leukocytes. Since ETB/ETBP are chemotactic for leukocytes and capable of inducing proliferation, cytokine and chemokine production, expression of adhesion molecules and apoptosis in renal cells and leukocytes, the early presence of these proteins could be a relevant event before and during antigen-antibody interaction takes place in renal tissues. PMID- 17139189 TI - Amelioration of established diabetic nephropathy by combined treatment with SMP 534 (antifibrotic agent) and losartan in db/db mice. AB - BACKGROUND/AIMS: Diabetic nephropathy is the main cause of end-stage renal disease. Previously we have demonstrated that SMP-534 (an antifibrotic agent) prevents the development of diabetic nephropathy in db/db mouse and that combined treatment with SMP-534 and losartan (antihypertensive agents) markedly prevents the development of diabetic nephropathy compared with single treatment. SMP-534 or losartan was prophylactically administered to db/db mice before the onset of diabetic nephropathy. In the present study, we evaluated the efficacy of combined treatment when administration was started after the onset of diabetic nephropathy. METHODS: db/db mice were raised untreated until 17 weeks of age, by which time increase of urinary albumin was noted, and then treated with SMP-534 and/or losartan for another 8 weeks. Biochemical and histological analyses were performed at 25 weeks of age. RESULTS: Combined treatment with SMP-534 and losartan markedly prevented the increase of urinary albumin and ameliorated the progression of mesangial matrix expansion, even when administration was started long after the increase of urinary albumin. CONCLUSION: The study results indicate that a combination of SMP-534 and losartan might be a valuable therapeutic approach for the treatment of diabetic nephropathy even when administration is started after the onset of diabetic nephropathy. PMID- 17139190 TI - Changes in osmolal gap and osmolality in children with chronic and end-stage renal failure. PMID- 17139191 TI - Healthy aging is associated with unaltered production of immunoreactive growth hormone but impaired neuroimmunomodulation. AB - BACKGROUND: Both endocrine and immune systems are continuously remodeled during aging. OBJECTIVE: Here, we investigated to what extent adrenal and somatosenescence are associated reciprocal changes in the immune system during strictly healthy aging. METHODS: Forty-six elderly subjects and 33 young adults were recruited according to the health criteria of the SENIEUR protocol. Peripheral blood mononuclear cells were isolated and stimulated in vitro with lipopolysaccharide or phytohemagglutinin to assess the production of immunoreactive growth hormone (GH). Peripheral sensitivity to steroids was assessed in vitro by dexamethasone-, cortisol- or dehydroepiandrosterone (DHEA) induced inhibition of T-cell proliferation. DHEA and GH levels were measured by radioimmunoassays. RESULTS: Healthy elderly had lower salivary DHEA and serum GH levels (somatosenescence). They presented reduced T-cell sensitivity to dexamethasone but similar cellular sensitivities to cortisol and DHEA. Their cells produced similar levels of immunoreactive GH compared to the cells of young adults. CONCLUSIONS: These data indicate that healthy aging is associated with adrenal and somatosenescence as well as impaired neuroendocrine immunoregulation at the level of the lymphocyte. In addition, somatosenescence may not be associated with a reciprocal decline in immunoreactive GH. PMID- 17139192 TI - Co-administration of acetyl-11-keto-beta-boswellic acid, a specific 5 lipoxygenase inhibitor, potentiates the protective effect of COX-2 inhibitors in kainic acid-induced neurotoxicity in mice. AB - Cyclooxygenase (COX) and lipoxygenase (LOX) are responsible for the metabolism of arachidonic acid into inflammatory metabolites, prostaglandins and leukotrienes, respectively. The upregulation of these enzymes in the central nervous system has been demonstrated to be responsible for the increased neuronal vulnerability to degeneration. Kainic acid, a glutamate receptor agonist and responsible for neuronal excitotoxicity and oxidative damage via different mechanisms, is capable of stimulating mRNA of both COX-2 and 5-LOX in the brain. The present study was designed to study the effects of COX inhibitors (indomethacin, nimesulide, rofecoxib) and a 5-LOX inhibitor (acetyl-11-keto-beta-boswellic acid; AKBA) and the combination of these inhibitors (dual inhibition) on kainic acid induced excitotoxicity and oxidative and nitrosative damage in mice. The results from the present study indicated that AKBA, indomethacin, and nimesulide per se did not produce any change in the behavioural parameters after kainic acid administration; however, rofecoxib per seproduced a significant increase in the latency of clonic (seizure-like) movement and a decrease in mortality rate as compared with kainic acid treated animals. In combination studies AKBA, rofecoxib, and nimesulide produced a more pronounced effect than either of these drugs alone. Further, the effect of AKBA combined with rofecoxib was significantly more marked when compared with AKBA combined with nimesulide. Besides this, identical results were found for the effect of these agents and their combination against oxidative damage induced by kainic acid. These findings indicate the potential role of COX-2 inhibitors and also their combination with the 5-LOX inhibitor in kainic acid induced excitotoxicity and oxidative damage by virtue of their antioxidant effect and suggest the need for the development of dual inhibitors for the treatment of neuronal excitotoxicity. PMID- 17139193 TI - Novel approaches to treatment of autoimmune neuroinflammation and lessons for drug development. AB - Drug development, and especially that intended for central nervous system (CNS) disorders, still poses a challenge. We investigated both the use of bifunctional compounds designed for multiple targeting and enhanced CNS permeability, and of recombinant alpha-fetoprotein (AFP), a natural pregnancy-associated immunomodulating protein for the treatment of CNS inflammation. Bifunctional compounds showed a novel pharmacokinetic profile due to the conjugation, yet retained, and even improved pharmacodynamics. AFP was well tolerated and decreased various aspects of neuroinflammation, including disease severity, axonal loss and damage, T-cell reactivity, and antigen presentation. Our results show that both strategies may serve as future drug modalities. PMID- 17139194 TI - Distinct effects of z-335, a new thromboxane A2 receptor antagonist, on rabbit platelets and aortic smooth muscle. AB - The effect of a novel thromboxane A2 receptor (TP) antagonist, (+/-)-sodium[2-(4 chlorophenylsulfonylaminomethyl)- indan-5-yl]acetate monohydrate (Z-335), on the U46619-induced responses was compared between rabbit platelets and aorta. Z-335 inhibited platelet shape change induced by U46619 with higher efficacy than SQ29548, a common TP antagonist. The U46619-induced platelet aggregation was inhibited by Z-335 in a noncompetitive manner, while it was competitively inhibited by SQ29548. Z-335 inhibited U46619-induced vasoconstriction of rabbit aorta with higher efficacy than SQ29548. The pA2 value of Z-335 in aortic vasoconstriction was significantly higher than in platelet shape change. The competitive binding study showed the higher pKi value of Z-335 against [3H] SQ29548 binding in rabbit aortic smooth muscle cells than in platelets. These data suggest that Z-335 has useful characteristics of TP antagonism. PMID- 17139195 TI - Glutathione-S-transferase pi expression in toxic epidermal necrolysis: a marker of putative oxidative stress in keratinocytes. AB - BACKGROUND: Toxic epidermal necrolysis (TEN) is a dramatic drug-induced emergency related to extensive destruction of the epidermis. There is evidence that its pathomechanism involves impaired detoxication of xenobiotics. Glutathione-S transferase pi (GST-pi) is a phase II detoxifying enzyme involved in drug metabolization by human keratinocytes. METHOD: Immunohistochemistry was performed in order to assess the expression of GST-pi in keratinocytes of TEN, other cutaneous adverse drug reactions and bullous pemphigoid. RESULTS: GST-pi was disclosed in the involved epidermis of 16/16 TEN patients. It was present in the cytoplasm of suprabasal keratinocytes. GST-pi was also expressed in the clinically uninvolved skin in a majority (8/12) of TEN patients. By contrast, it was rarely and poorly expressed in the other tested dermatoses. CONCLUSION: The pathomechanism of TEN is not related to an impaired quantitative expression of GST-pi. GST-pi expression is an early event in TEN. As oxidative stress is a major inducer of GST-pi, this mechanism might be involved in TEN. Its GST-pi expression mainly restricted to the suprabasal keratinocytes suggests that the pathomechanisms leading to keratinocyte death in TEN are distinct at different levels of the epidermis. PMID- 17139196 TI - Overexpression of human chorionic gonadotropin beta genes 3, 5 and 8 in tumor tissue and urinary cells of bladder cancer patients. AB - OBJECTIVE: Human chorionic gonadotropin (hCG) is a marker of trophoblastic tumors, and the serum concentration of the free beta-subunit (hCGbeta) is an independent prognostic marker in several nontrophoblastic cancers. hCGbeta is encoded by six genes, of which the type II genes (hCGbeta 3/9, 5 and 8) are thought to be upregulated in relation to type I genes (hCGbeta 6/7) in cancer. METHOD: We developed a novel quantitative RT-PCR method for the quantification of the relative expression levels of the two groups of hCGbeta genes and analyzed 28 bladder tumors and 15 urine samples. RESULTS: We found a higher relative expression level of type II genes in malignant compared with benign urothelia (p = 0.016) and in exfoliated urinary cells from cancer patients compared with those from benign controls (p = 0.026). The expression level was increasing with higher stage (p = 0.014) and grade (p = 0.001) and tended to be higher in relapsing tumors (p = 0.059). CONCLUSION: The increased hCGbeta concentrations in body fluids of patients with aggressive bladder cancer may be due to overexpression of type II genes. Quantification of the relative mRNA expression levels of the hCGbeta type I and II genes in urine cells should be further studied as a potential noninvasive tool for the diagnosis and follow-up of bladder cancer. PMID- 17139197 TI - A model for dealing with sudden death. AB - The purpose of this study was to attempt to provide health professionals working in emergency departments with guidelines to use in order to ensure the therapeutic management of the dead or dying client, the suddenly bereaved families, and fellow colleagues (health professionals). A 4-year action research study was undertaken involving semistructured interviews with health professionals (doctors and nurses) working in selected Level I emergency departments, suddenly bereaved families belonging to a local bereavement support group, and mortuary staff in KwaZulu-Natal, South Africa. Further analysis of the categories and themes that emerged from the interviews informed the development of the Dealing with Sudden Death Model, a family information pamphlet, a preparation checklist, and an incident evaluation checklist. PMID- 17139198 TI - An exploration of the good death. AB - For many patients with a life-threatening illness, modern hospitals prevent a good death. When aggressive treatment is selected for a disease process with a remote cure, nurses engage in patient care that is psychologically exhausting and ethically demoralizing. Nursing is well positioned to lead a paradigm shift regarding end-of-life care. The concept of good death is explored through sociology, Christian theology, medicine, and nursing. Of the many determinants for a good death, the ones that transcend the disciplines include making adequate preparations, experiencing no unpleasant symptoms, having someone by one's side, and being spiritually whole. Empirical indicators for measuring a good death are also explored. PMID- 17139199 TI - Patients and families go on with life when a terminal illness enforces hospitalization: an interpretive phenomenological study. AB - This interpretive phenomenological study explored patients' with a terminal illness and their family members' experiences of hospital end-of-life care. Findings show that patients and families led unique lives, and even when a terminal illness intruded and enforced a hospital stay, they held on to familiar practices. Patients and family members felt best cared for when healthcare providers supported them in their endeavors to go on with their usual life as much as possible. To achieve this, professionals are challenged to recognize patients and family members as persons and to integrate the latter's lived experiences in their care interventions. PMID- 17139200 TI - A universal and fundamental nursing philosophy. PMID- 17139201 TI - Chinn's editorial, "Physiology and the idea of holism". PMID- 17139202 TI - In our grandmother's footsteps: perceptions of being strong in African American women with HIV/AIDS. AB - One of the most significant challenges facing the health of black women in the 21st century is the growing numbers of human immunodeficiency virus/acquired immunodeficiency disease (HIV/AIDS) infections. An ethnographic study of African American mothers living with HIV/AIDS revealed that they believed in a tradition and heritage of strength that fostered their survival during difficult life experiences such as living and mothering with HIV/AIDS. They enacted this strength in culturally significant ways. This article discusses the importance of recognizing and supporting cultural strengths of African American women to help manage illness, while remaining cognizant of the context of oppression, discrimination, and stigma that distort cultural traditions and instead penalize women when they are ill. PMID- 17139203 TI - Life changing words: women's responses to being diagnosed with HIV infection. AB - In this longitudinal study, we investigated the diagnosis experiences of 55 human immuno-deficiency virus (HIV)-infected women. Women's immediate reactions upon hearing that they were infected with HIV were devastation, shock, and indignation. Long-term responses included depression, submersion of the HIV infection diagnosis, escalated drug and alcohol use, shame, and suicidality. It was usually months and sometimes years before women could extricate themselves from these patterns of response. It is critical to make HIV infection diagnosis the first intervention in a protocol of seamless support that sees women through the initial trauma of being diagnosed until longer term primary care and social services can be activated. PMID- 17139204 TI - A sudden, life-threatening medical crisis: the family's perspective. AB - Each year millions of Americans are hospitalized suddenly with critical injuries or life-threatening medical emergencies. The families of these patients begin a roller coaster ride characterized by terrified waiting and lack of control. All the while, they are uncertain whether their loved one will survive until the ride is over. This is their story. Phenomenologic interviews with 6 family members identified 10 theme clusters that describe the experience. Specific implications for the nursing care of these families are discussed. PMID- 17139205 TI - A situation-specific theory of Caucasian cancer patients' pain experience. AB - Situation-specific theories can explain a population's unique health/illness experience with a limited scope of generalizability, but with more specificity that can provide a more close and comprehensive view on and explanation of a nursing phenomenon. In this article, a situation-specific theory explaining the unique cancer pain experience of Caucasian cancer patients is presented, along with the theory development process that was taken to develop the theory. First, the method used to develop the theory is concisely described. Then, the theory development process is described step by step. Finally, the situation-specific theory that was developed through the process is presented. PMID- 17139206 TI - The recovery process in major depression: an analysis employing Meleis' transition framework for deeper understanding as a foundation for nursing interventions. AB - The number of persons with mental illness is increasing globally; despite this fact, nursing research on research-based interventions to prevent or minimize illness and increase quality of life is sparse. The purpose of this secondary analysis of men and women recovering from major depression (N = 25) was to gain a deeper understanding of the concept of transition in the recovery process associated with major depression as well as to develop and suggest nursing interventions that support the recovery process. The transition framework was useful, as it was describing the transition process as fluid, going back and forth, which was confirmed by the respondents' statements. Transition planning is a feasible way of supporting both the recovery process and health promotion, thus laying the foundations for a good quality of life. PMID- 17139207 TI - Enriching the portrait: methodological triangulation of life-closing theory. AB - Some researchers promote a triangulated or multiple-methods approach in studies that examine complex human phenomenon. Nonetheless, problems abound in combining 2 dissimilar data sets from 2 different methodologies. One way that numerical and textual data can be merged is through a research method called statistical triangulation. The purpose of this article was to describe the statistical triangulation of a simultaneous, between-methods causal model and grounded theory investigation that explained psychological adaptation in dying persons. The statistical triangulation involved the analysis of selected patterns of dying from the grounded theory study with the variables used in the causal model investigation. A 1-way analysis of variance confirmed that the variables of social support, physical function, and religious preference impacted end-of-life patterns. Post hoc comparisons validated the conceptualization of 3 patterns of dying (becoming, anguishing/agonizing, avoiding) that emerged from the qualitative data. Although there are limitations to this analysis, statistical triangulation shows promise as a research method for enriching qualitative description. PMID- 17139208 TI - A transactional model of cancer family caregiving skill. AB - Family caregivers of persons receiving treatment of cancer must acquire illness related skills not usually found among nonprofessional caregivers. Although research is needed to understand the skill acquisition process, no models of family caregiving skill exist to guide such research. The purpose of this study was to develop a model of caregiving skill through qualitative research. Participants were 44 caregiver/patient dyads and 63 individual caregivers. Data were collected using a semi-structured interview. A transactional model of family caregiving skill was developed that includes the concepts of demands of the illness situation, patterns of care, caregiver responses, and patient responses. PMID- 17139209 TI - New rod-plate anterior instrumentation for thoracolumbar/lumbar scoliosis: biomechanical evaluation compared with dual-rod and single-rod with structural interbody support. AB - STUDY DESIGN: A new rod-plate anterior implant was designed to provide plate fixation at the cephalad and caudal-end segments of a 5-level anterior spine construct. Biomechanical testing was performed on calf spines instrumented with 5 segment anterior scoliosis constructs. OBJECTIVES.: To analyze the initial and post-fatigue biomechanical performance of the new implant, and compare it to an anterior dual-rod construct and a single-rod construct with interbody cages. SUMMARY OF BACKGROUND DATA: Using single-rod anterior instrumentation for thoracolumbar and lumbar scoliosis, an unacceptable incidence of loss of correction, segmental kyphosis, and pseudarthrosis has been reported. Inadequate construct stiffness due to early postoperative bone-screw interface failure, especially at cephalad and caudal-end vertebrae, has been implicated as the cause of these complications. METHODS: Thirty calf spines were instrumented over 5 segments with: (1) single-rod augmented with rod-plate implants, (2) dual-rod construct, and (3) single-rod with titanium mesh cages. Stiffness in flexion extension and lateral bending modes was determined initially and post-cyclical loading by measuring segmental range of motion (ROM). Post-fatigue screw pullout tests were also performed. RESULTS: In lateral bending, the caudal-end segmental ROM for rod-plate construct was 54% less than single-rod with cages construct (P < 0.05), with no difference between rod-plate and dual-rod constructs. In flexion extension, the rod-plate construct showed 45% to 91% (initial test) and 84% to 90% (post-fatigue) less ROM than the single-rod with cages construct (P < 0.001). Again, there was no difference between rod-plate and dual-rod constructs at the cephalad and caudal-end segments. Post-fatigue screw pullout strengths of the rod plate construct were significantly greater than those of the dual-rod and single rod with cages constructs (P < 0.05). CONCLUSIONS: The rod-plate construct was significantly stiffer and provided greater stability of bone-screw interface than the single-rod with cages construct. It achieved similar stiffness and improved bone-screw interface stability compared to dual-rod construct. PMID- 17139210 TI - Biomechanical evaluation of conventional internal contemporary spinal fixation techniques used for stabilization of complete sacroiliac joint separation: a 3 dimensional unilaterally isolated experimental stiffness study. AB - STUDY DESIGN: Comparative 3-dimensional biomechanical testing. OBJECTIVE: To compare 5 fixation techniques, 3 using screws or screw and plates and 2 spinal, used for stabilization of complete unilateral sacroiliac dislocation in composite models. SUMMARY OF BACKGROUND DATA: Harrington compression rods have been used for posterior iliosacral stabilization. Recently, the use of compact spinal instrumentation has been introduced for stabilization of iliosacral joint separation to achieve immediate and permanent stability, allowing early mobilization. To the authors' knowledge, no comparative mechanical studies between commonly used internal fixation techniques and contemporary spinal instrumentation have been performed. METHODS: Fifteen identical composite models of the left hemipelvis and sacrum were used to simulate consistently the "worst case scenario" of complete unilateral sacroiliac dislocation. Subgroups of 3 models each were used to apply 5 (A-E) alternative fixation iliosacral joint fixation techniques: 1 multiaxial 7.5 mm Cotrel-Dubousset screw inserted in the posterior superior iliac spine and connected with a long Cotrel-Dubousset horizontal rod with 6.5 mm multiaxial Cotrel-Dubousset screws inserted bilaterally in the S1 pedicles (technique A); 1 multiaxial 7.5 mm Cotrel Dubousset titanium pedicle screw inserted in the posterior superior iliac spine and connected with a short horizontal Cotrel-Dubousset-rod to a 6.5 mm multiaxial Cotrel-Dubousset-screw inserted to the ipsilateral S1 pedicle (technique B); 1, 6.5 mm cancellous AO-screw (technique C); 2, 6.5 mm cancellous AO screws (technique D); and 2 dynamic stainless steel compression plates (technique E) placed anteriorly. Constructs were biomechanically tested. The ilium was unilaterally rigidly fixed, the sacrum was put horizontal in the mediolateral direction with a forward tilt of 30 degrees (close to physiologic conditions) in the sagittal plane, and a vertical quasi-static compressive load ranging from 0 to 500 N was applied on the endplate of S1, reproducing a "worst case" loading scenario. Construct stiffness, frontal plus sagittal kinematics, and iliosacral joint gap size for all 5 techniques were measured. RESULTS: The construct stiffness (N/mm +/- standard deviation) ranged for model: A, 121 +/- 18; B, 78 +/ 10; C, 168 +/- 13; D, 193 +/- 42; and E, 145 +/- 4. All other parameters exhibited minor variations between the different techniques of fixation: at the 400 N load level, the maximum iliosacral gap globally ranged 0.9-2.8 mm, the maximum mediolateral sacral tilt ranged 1.3-2.4 degrees, and the maximum anteroposterior sacral tilt ranged 0.6-3.0 degrees. CONCLUSIONS: The iliosacral fixation with 2 6.5 mm AO-cancellous screws for complete sacroiliac dislocation demonstrated the highest stiffness and the short spinal instrumentation the poorest stiffness. All other fixation techniques could be generally considered of equivalent stability value. PMID- 17139211 TI - Static versus dynamic loading in the mechanical modulation of vertebral growth. AB - STUDY DESIGN: Measures of absolute and relative growth modulation were used to determine the effects of static and dynamic asymmetric loading of vertebrae in the rat tail. OBJECTIVES: To quantify the differences between static and dynamic asymmetric loading in vertebral bone growth modulation. SUMMARY OF BACKGROUND DATA: The creation and correction of vertebral wedge deformities have been previously described in a rat-tail model using static loading. The effects of dynamic loading on growth modulation in the spine have not been characterized. METHODS: A total of 36 immature Sprague-Dawley rats were divided among four different groups: static loading (n = 12, 0.0 Hz), dynamic loading (n = 12, 1.0 Hz), sham operated (n = 6), and growth controls (n = 6). An external fixator was placed across the sixth and eighth caudal vertebrae as the unviolated seventh caudal vertebra was evaluated for growth modulation. Static or dynamic asymmetric loads were applied at a loading magnitude of 55% body weight. After 3 weeks of loading, growth modulation was assessed using radiographic measurements of vertebral wedge angles and vertebral body heights. RESULTS: The dynamically loaded rats had a final average wedge deformity of 15.2+/- 6.4 degrees, which was significantly greater than the statically loaded rats whose final deformity averaged 10.3 degrees +/- 3.7 degrees (P < 0.03). The deformity in both groups was statistically greater than the sham-operated (1.1+/- 2.0 degrees) and growth control rats (0.0+/- 1.0 degrees) (P < 0.001). The longitudinal growth was significantly lower on the concavity compared with the convexity in both the dynamically (0.34 +/- 0.23 mm vs. 0.86 +/- 0.23 mm) and statically (0.46 +/- 0.19 mm vs. 0.83 +/- 0.32 mm) loaded rats (P < 0.001). These growth rates were significantly less than the sham operated and growth control rats (P < 0.001). CONCLUSIONS: A variety of fusionless scoliosis implant strategies have been proposed that use both rigid and flexible implants to modulate vertebral bone growth. The results from this study demonstrate that dynamic loading of the vertebrae provides the greatest growth modulation potential. PMID- 17139212 TI - Gravity line analysis in adult volunteers: age-related correlation with spinal parameters, pelvic parameters, and foot position. AB - STUDY DESIGN: Prospective radiographic and forceplate analysis in adult volunteers. OBJECTIVE: Assess gravity line (GL) location and foot position regarding anatomic spinal structures to evaluate key correlations and age-related changes in balance. SUMMARY OF BACKGROUND DATA: Global spinal balance is commonly assessed by the C7 plumbline. This radiographic parameter does not offer information on foot position or forces transmitted, and poor correlation with the true GL has been demonstrated. METHODS: A total of 75 asymptomatic adult volunteers were equally distributed into three age groups. Full length, free standing spine radiographs were obtained with simultaneous acquisition of GL and feet location (forceplate). GL and heels were projected on each radiograph to compute their distance from anatomic entities and to investigate correlations with radiologic parameters and age-related changes. RESULTS: In this study group, advancing age led to a significant increase in thoracic kyphosis. The plumbline from C7 shifted anteriorly with age. In the sagittal plane, the GL was anterior to the vertebral column for all groups. With age, the GL location regarding the heels remained constant, while the pelvis moved posteriorly toward the heels and underwent a small retroversion (increasing pelvic tilt). The acetabulum was the most reliable radiographic marker of the GL location. CONCLUSIONS: This quantitative study in volunteers reveals clear age-related changes in the spino pelvic association and offers quantitative support to the "cone of economy" concept proposed by Dubousset. The pelvis can be seen as a regulator to help maintain a rather fixed GL-heel association with age-related changes in the spinal column. Further study in patients suffering from deformity can confirm the importance of radiographic-gravity line correlations and enhance our understanding of optimal balance. PMID- 17139213 TI - Prevalence, risk factors, and preference-based health states of low back pain in a Turkish population. AB - STUDY DESIGN: A community-based cross-sectional survey was conducted using face to-face interviews. OBJECTIVES: To estimate the point, 12-month period, and lifetime prevalence of low back pain (LBP) in the general population of Turkey, and to determine the factors associated with LBP. We also aim to determine self reported health states differences between patients and the general population by the EuroQol-5D & Visual Analogue Scale (EQ5D-VAS). SUMMARY OF BACKGROUND DATA: LBP as a major public health problem has been unquestionably the most serious pain problem. LBP generally appears between 35 and 64 years of age. Little was known about the prevalence and determinants of LBP among populations from developing areas; information mostly comes from industrialized countries. This epidemiologic study reported the prevalence of LBP in the general population in Turkey. In Turkey, community-based trials on the prevalence of LBP were scarce. METHODS: A total of 7,000 participants selected by randomly sampling from seven regions of Turkey. A comprehensive questionnaire on demographics, physical and emotional stress, LBP, and EQ5D-VAS was administered to 4,990 men and women (age range, 18-70 years). RESULTS: The lifetime, 12-month period, and point prevalence of LBP were 44.1%, 34.0%, and 19.7%, respectively. The prevalence of LBP in women was consistently higher than in men across all age groups. The proportion of specific LBP was 10.2%, and most common etiology of specific LBP was disc hernia. In regression analysis, occupation, physical, and emotional stress showed significant associations with having LBP. The EQ5D-VAS scores of patients with LBP were significantly lower than self-reported assessments of the population without LBP (64.6, 63.0 vs. 73.8, P < 0.05). CONCLUSIONS: In comparison with other developing countries, the prevalence of LBP is higher in Turkey and approximates prevalence estimates of LBP in developed countries. LBP is associated with occupation. Smoking is not associated with having LBP. This study also shows that LBP influences disability and quality of life. PMID- 17139214 TI - Mathematical calculation of pedicle subtraction osteotomy size to allow precision correction of fixed sagittal deformity. AB - STUDY DESIGN: This is a retrospective review of 15 consecutive fixed sagittal plane deformity patients who have undergone pedicle subtraction osteotomies. The focus of this article is the application of a trigonometric equation that calculates the degree of correction needed to achieve sagittal balance. The intraoperative predictive accuracy and clinical radiographic results of using this mathematical equation are discussed. OBJECTIVE: The need for a precise and reproducible planning tool for the correction of sagittal imbalance prompted us to apply a simple trigonometric equation to achieve the desired sagittal alignment of the spine. SUMMARY OF BACKGROUND DATA: Establishing sagittal balance has been widely recognized as one of the most important parameters in optimizing outcomes for spinal reconstruction patients. Preoperative planning for sagittal plane correction in adult spinal deformity has traditionally been done by estimation or with cumbersome film cutouts. To our knowledge, there has not been a consistent method of calculating the exact number of degrees needed to reestablish spinal balance. METHODS: Patients' C7 plumb lines are measured on a 36-inch radiograph to assess the degree of sagittal imbalance and determine how many degrees of correction (and subsequent millimeters of wedge resection) are needed. Applying a basic trigonometric formula for the tangent to the sagittal alignment is used to do this. RESULTS: We have used this technique reliably in a series of 15 consecutive patients to reestablish sagittal balance. The predicted degree of correction was compared to the achieved degree of correction at the site of the osteotomy. This comparison was accurate to within 3 degrees (the standard error of measurement for the method of Cobb) in all cases except 2. CONCLUSIONS: By using a simple mathematical equation, one can reliably determine the degree of pedicle subtraction osteotomy needed for correction of sagittal deformity. This technique is reproducible and has led to successful clinical outcomes. PMID- 17139215 TI - Quadriparesis due to intraspinal cyst after failed posterior occipitocervical fusion in a patient with athetoid cerebral palsy. AB - STUDY DESIGN: A case report and review of the literature are presented. OBJECTIVES: To describe the clinical course and treatment of a patient with athetoid cerebral palsy who had quadriparesis due to an intraspinal cyst, following a failed occipitocervical fusion using sublaminar wires and rods. SUMMARY OF BACKGROUND DATA: Intraspinal cyst as a cause of quadriparesis in a patient with athetoid cerebral palsy is extremely rare. To our knowledge, there have been no publications on this issue. METHODS: A 60-year-old man with athetoid cerebral palsy and a history of posterior occipitocervical fusion presented with quadriparesis. Salvage surgery for cervical myelopathy and pseudarthrosis was performed with laminectomy and rearthrodesis using the pedicle screw system. An intraspinal cyst was identified as the main cause of the paraparesis. RESULTS: Solid bony fusion and the improvement of paraparesis were achieved 2 years and 1 month after the surgery. He is now able to feed himself and to walk with a cane, both without assistance. CONCLUSION: A physician managing patients with athetoid cerebral palsy should always be aware that an intraspinal cyst in the cervical spine may be the cause of cervical myelopathy. PMID- 17139217 TI - Circumferential fusion improves outcome in comparison with instrumented posterolateral fusion: long-term results of a randomized clinical trial. AB - STUDY DESIGN: Prospective randomized clinical study with a 5- to 9-year follow-up period. OBJECTIVE: The aim of the present study was to analyze the long-term outcome with respect to functional disability, pain, and general health of patients treated by means of circumferential lumbar fusion in comparison with those treated by means of instrumented posterolateral lumbar fusion. SUMMARY OF BACKGROUND DATA: Circumferential fusion has become a common procedure in lumbar spinal fusion both as a primary and salvage procedure. However, the claimed advantages of circumferential fusion over conventional posterolateral fusion lack scientific documentation. (The primary report with a 2-year follow-up has been published in Spine in 2002.) METHODS: From April 1996 to November 1999, a total of 148 patients (mean age, 45 years) with severe chronic low back pain were randomly selected for either posterolateral lumbar fusion (titanium Cotrel Dubousset) or circumferential lumbar fusion (instrumented posterolateral fusion with anterior intervertebral support by a Brantigan cage). The primary outcome measure was the Dallas Pain Questionnaire (DPQ). The secondary outcome measures were the Oswestry Disability Index, the SF-36 instrument, and the Low Back Pain Rating Scale. All measures assessed the endpoint outcomes at 5 to 9 years after surgery. RESULTS: The available response rate was 93%. The circumferential group showed a significantly better improvement (P < 0.05) in comparison with the posterolateral group with respect to all four DPQ categories: daily activities, work/leisure, anxiety/depression, and social interest. The Oswestry Disability Index supported these results (P < 0.01). General health, as assessed by means of the SF-36, also showed significantly better physical health (P < 0.01) in the circumferential group, whereas no significant difference was found with respect to mental health compared with the posterolateral group. The circumferential group experienced significantly less back pain (P < 0.05) in comparison with the posterolateral group. In regard to leg pain, no significant difference was found. CONCLUSION: Circumferential lumbar fusion demands more extensive operative resources compared with posterolateral lumbar fusion. However, 5 to 9 years after surgery, the circumferentially fused patients had a significantly improved outcome compared with those treated by means of posterolateral fusion. These new results not only emphasize the superiority of circumferential fusion in the complex pathology of the lumbar spine but are also strongly supported in all of the validated questionnaires used in the study. PMID- 17139218 TI - Treatment of traumatic thoracolumbar spine fractures: a multicenter prospective randomized study of operative versus nonsurgical treatment. AB - STUDY DESIGN: Multicenter prospective randomized trial. OBJECTIVE: To test the hypotheses that thoracolumbar AO Type A spine fractures without neurologic deficit, managed with short-segment posterior stabilization will show an improved radiographic outcome and at least the same functional outcome as compared with nonsurgically treated thoracolumbar fractures. SUMMARY OF BACKGROUND DATA: There are various opinions regarding the ideal management of thoracolumbar Type A spine fractures without neurologic deficit. Both operative and nonsurgical approaches are advocated. METHODS: Patients were randomized for operative or nonsurgical treatment. Data sampling involved demographics, fracture classifications, radiographic evaluation, and functional outcome. RESULTS: Sixteen patients received nonsurgical therapy, and 18 received surgical treatment. Follow-up was completed for 32 (94%) of the patients after a mean of 4.3 years. At the end of follow-up, both local and regional kyphotic deformity was significantly less in the operatively treated group. All functional outcome scores (VAS Pain, VAS Spine Score, and RMDQ-24) showed significantly better results in the operative group. The percentage of patients returning to their original jobs was found to be significantly higher in the operative treated group. CONCLUSIONS: Patients with a Type A3 thoracolumbar spine fracture without neurologic deficit should be treated by short-segment posterior stabilization. PMID- 17139219 TI - Mechanisms of anular failure resulting from excessive intradiscal pressure: a microstructural-micromechanical investigation. AB - STUDY DESIGN: Microstructural/micromechanical investigation of pathways of anular wall disruption. OBJECTIVE: To investigate the fundamental mechanisms by which both intralamellar and interlamellar relationships are disrupted by nuclear pressurization. SUMMARY OF BACKGROUND DATA: Understanding how anular failure might occur following increased nuclear pressurization requires an experimental approach that avoids artifactual injury to the anulus but reveals structural disruption resulting directly from the pressurization event. METHODS: Bovine motion segments were subjected to internal pressurization using a novel "through vertebra" gel injection method. Intralamellar and interlamellar sections were deliberately chosen so as to expose systematic patterns of structural disruption resulting from the pressurization event. This microdisruption was investigated using a novel method that combined microtensile manipulation and simultaneous differential contrast imaging of the fully hydrated unstained sections. RESULTS: The inner anulus was most severely disrupted, the middle regions developed a series of regular clefts along axes of weakness within the in-plane arrays, with only mild array disruption occurring in the outer regions. CONCLUSIONS: A mechanism is proposed whereby an anular failure pathway, driven by hydrostatic nuclear pressure, could track through the complex anular structures via a set of disruptive events causing weakening of both the in-plane and interlamellar junction interconnections. PMID- 17139220 TI - Injection of nuclear factor-kappa B decoy into the sciatic nerve suppresses mechanical allodynia and thermal hyperalgesia in a rat inflammatory pain model. AB - STUDY DESIGN: In vitro and in vivo study of a rat inflammatory pain model using nuclear factor-kappa B decoy. OBJECTIVES: To investigate transduction efficiency of nuclear factor-kappa B decoy into dorsal root ganglion, both in vivo and in vitro, and to assess the suppression of inflammatory pain by nuclear factor-kappa B decoy. SUMMARY OF BACKGROUND DATA: Transcription factor nuclear factor-kappa B is reported to play a crucial role in regulating pro-inflammatory cytokine gene expression. We hypothesized that inhibiting nuclear factor-kappa B gene expression with nuclear factor-kappa B decoy may suppress inflammatory pain. METHODS: Nuclear factor-kappa B decoy-fluorescein isothiocyanate (FITC) was induced in explant culture, endoneurally injected into the sciatic nerve, and its transduction efficiency into dorsal root ganglion measured. For behavioral testing, 12 rats received plantar injections of complete Freund's adjuvant and were divided into 3 groups: decoy group, single endoneural injection of 10 microL of nuclear factor-kappa B decoy (n = 4); saline group, single endoneural injection of 10 microL of saline (n = 4); and naive group, untreated (n = 4). Behavioral testing was performed using von Frey filaments and a Hargreaves device with a heat source. RESULTS: Total transduction efficiency of nuclear factor kappa B decoy-FITC was 53.6% in vitro and 20.5% in vivo. No statistical differences were observed with respect to types of cell size distributions of all FITC-positive neurons. In behavioral testing, withdrawal latencies or thresholds significantly differed between the decoy group and the saline group from 2 to 14 days after surgery in the mechanical allodynia experiments, and from 2 to 3 days after surgery in the thermal hyperalgesia experiments. CONCLUSIONS: Nuclear factor-kappa B decoy was conveyed and transduced into dorsal root ganglion both in vivo and in vitro. Additionally, nuclear factor-kappa B decoy reduced mechanical allodynia and thermal hyperalgesia in the rat inflammatory pain model, suggesting that inhibition of nuclear factor-kappa B with nuclear factor-kappa B decoy may represent a key mechanism for mediating inflammation or reducing inflammatory pain. PMID- 17139221 TI - Effects of growth differentiation factor-5 on the intervertebral disc--in vitro bovine study and in vivo rabbit disc degeneration model study. AB - STUDY DESIGN: In vitro studies on the effects of recombinant human growth and differentiation factor-5 (rhGDF-5) on matrix metabolism of bovine intervertebral disc cells and an in vivo study on the effect of rhGDF-5 in the rabbit anular puncture model. OBJECTIVE: To determine the reparative capacity of rhGDF-5 on the intervertebral disc. SUMMARY OF BACKGROUND DATA: The in vitro and in vivo effects of rhGDF-5, a crucial protein in the developing musculoskeletal system, on repair of the degenerated intervertebral disc remain unidentified. METHODS: In vitro, bovine nucleus pulposus and anulus fibrosus cells were cultured with or without rhGDF-5 (100 or 200 ng/mL). On days 7, 14, and 21, the contents of deoxyribonucleic acid and proteoglycan, and the synthesis of proteoglycan and collagen were assessed. In vivo, 16 adolescent New Zealand white rabbits received anular punctures in 2 lumbar discs. Four weeks later, phosphate buffered saline or rhGDF-5 (10 ng, 1 and 100 mug) was injected into the nucleus pulposus. The rabbits were followed up for 16 weeks for disc height, magnetic resonance imaging, and histologic grading. RESULTS: In vitro, rhGDF-5 increased the deoxyribonucleic acid and proteoglycan contents of both cell types significantly after day 14. rhGDF-5 at 200 ng/mL significantly stimulated proteoglycan synthesis (nucleus pulposus: +138%, anulus fibrosus: +24%) and collagen synthesis (nucleus pulposus: +95%, anulus fibrosus: +23%) at day 21. In vivo, the injection of rhGDF-5 resulted in a restoration of disc height, improvement of magnetic resonance imaging scores, and histologic grading scores with statistical significance (P < 0.05-0.001). CONCLUSION: A single injection of rhGDF-5 has a reparative capacity on intervertebral discs, presumably based on its effects to enhance extracellular matrix production in vitro. PMID- 17139222 TI - Establishment of a novel intervertebral disc/endplate culture model: analysis of an ex vivo in vitro whole-organ rabbit culture system. AB - STUDY DESIGN: Ex vivo in vitro study evaluating a novel intervertebral disc/endplate culture system. OBJECTIVES: To establish a whole-organ intervertebral disc culture model for the study of disc degeneration in vitro, including the characterization of basic cell and organ function. SUMMARY OF BACKGROUND DATA: With current in vivo models for the study of disc and endplate degeneration, it remains difficult to investigate the complex disc metabolism and signaling cascades. In contrast, more controlled but simplified in vitro systems using isolated cells or disc fragments are difficult to culture due to the unconstrained conditions, with often-observed cell death or cell dedifferentiation. Therefore, there is a demand for a controlled culture model with preserved cell function that offers the possibility to investigate disc and endplate pathologies in a structurally intact organ. METHODS: Naturally constrained intervertebral disc/endplate units from rabbits were cultured in multi-well plates. Cell viability, metabolic activity, matrix composition, and matrix gene expression profile were monitored using the Live/Dead cell viability test (Invitrogen, Basel, Switzerland), tetrazolium salt reduction (WST-8), proteoglycan and deoxyribonucleic acid quantification assays, and quantitative polymerase chain reaction. RESULTS: Viability and organ integrity were preserved for at least 4 weeks, while proteoglycan and deoxyribonucleic acid content decreased slightly, and matrix genes exhibited a degenerative profile with up regulation of type I collagen and suppression of collagen type II and aggrecan genes. Additionally, cell metabolic activity was reduced to one third of the initial value. CONCLUSIONS: Naturally constrained intervertebral rabbit discs could be cultured for several weeks without losing cell viability. Structural integrity and matrix composition were retained. However, the organ responded to the artificial environment with a degenerative gene expression pattern and decreased metabolic rate. Therefore, the described system serves as a promising in vitro model to study disc degeneration in a whole organ. PMID- 17139223 TI - Rapid atrophy of the lumbar multifidus follows experimental disc or nerve root injury. AB - STUDY DESIGN: Experimental study of muscle changes after lumbar spinal injury. OBJECTIVES: To investigate effects of intervertebral disc and nerve root lesions on cross-sectional area, histology and chemistry of porcine lumbar multifidus. SUMMARY OF BACKGROUND DATA: The multifidus cross-sectional area is reduced in acute and chronic low back pain. Although chronic changes are widespread, acute changes at 1 segment are identified within days of injury. It is uncertain whether changes precede or follow injury, or what is the mechanism. METHODS: The multifidus cross-sectional area was measured in 21 pigs from L1 to S1 with ultrasound before and 3 or 6 days after lesions: incision into L3-L4 disc, medial branch transection of the L3 dorsal ramus, and a sham procedure. Samples from L3 to L5 were studied histologically and chemically. RESULTS: The multifidus cross sectional area was reduced at L4 ipsilateral to disc lesion but at L4-L6 after nerve lesion. There was no change after sham or on the opposite side. Water and lactate were reduced bilaterally after disc lesion and ipsilateral to nerve lesion. Histology revealed enlargement of adipocytes and clustering of myofibers at multiple levels after disc and nerve lesions. CONCLUSIONS: These data resolve the controversy that the multifidus cross-sectional area reduces rapidly after lumbar injury. Changes after disc lesion affect 1 level with a different distribution to denervation. Such changes may be due to disuse following reflex inhibitory mechanisms. PMID- 17139224 TI - Biomechanical evaluation of vertebroplasty and kyphoplasty with polymethyl methacrylate or calcium phosphate cement under cyclic loading. AB - STUDY DESIGN: We developed a new method to simulating in vivo dynamic loading as closely as possible, which allows comparison of kyphoplasty and vertebroplasty, as well as augmentation materials. OBJECTIVE: Special interest was given to calcium phosphate cement, which might fail due to its brittleness. SUMMARY OF BACKGROUND DATA: Vertebroplasty and kyphoplasty are, with limitations, 2 promising alternative techniques to augment osteoporotic vertebrae with polymethyl methacrylate or calcium phosphate cements. However, little is known about the fatigue characteristics of the treated vertebrae under cyclic loading. METHODS: Twenty-four intact, osteoporotic bi-segmental human specimens were divided into 4 groups: (1) vertebroplasty with polymethyl methacrylate, (2) kyphoplasty with polymethyl methacrylate, (3) kyphoplasty with calcium phosphate cement, and (4) untreated control group. After augmentation of the middle vertebrae, all specimens underwent 100,000 cycles of eccentric loading during which the specimen revolved around its longitudinal axis. Pre-loading and post loading radiographs, and subsidence measurements at different sites of the vertebrae were taken. The overall height was additionally determined every 20,000 cycles in the material testing machine. Finally, the specimens were cryosectioned to examine the cements. RESULTS: Loss of height progressed with strong individual differences in all groups, with an increasing number of load cycles up to median values of 2.8 mm for both augmented groups and 4.2 mm for the nonaugmented group. At the center of the upper endplate, subsidence in kyphoplasty was greater than in vertebroplasty, with little differences with respect to the kind of cement. The cryosections did not show any signs of fatigue in the polymethyl methacrylate, but small cracks were in the calcium phosphate. CONCLUSIONS: Vertebroplasty and kyphoplasty seem to be equivalent methods in strengthening osteoporotic vertebrae. However, these results cannot be transferred to the treatment of fractures with these methods. A "physiologic" loading situation was achieved by complex motion, including all combinations of flexion/extension with lateral bending during eccentric cyclic loading. PMID- 17139225 TI - Does minor trauma cause serious low back illness? AB - STUDY DESIGN: Prospective, 5-year, cohort study of working subjects. OBJECTIVES: To assess whether the occurrence of common minor trauma events affects the risk of developing serious low back pain (LBP) and LBP disability in subjects with and without degenerative changes to the lumbar spine. SUMMARY OF BACKGROUND DATA: Although some theories suggest that minor traumatic events in combination with preexisting degenerative changes commonly cause significant structural injury to spinal segments and serious LBP illness, no prospective data exist on the relationship of minor trauma, detailed structural changes, and outcome measures of serious LBP episodes and occupational disability. METHODS: Two hundred subjects without clinical LBP problems were recruited, and underwent baseline clinical and imaging studies. Every 6 months, subjects completed a scripted, algorithm-based interview assessing interval back pain episodes, severity, medical treatment, occupational disability, and the subject's perceived relation of this LBP episode to any preceding event. If a serious LBP episode clinically required a new magnetic resonance examination, the follow-up imaging was obtained and compared to baseline for interval changes. RESULTS: There was no association of minor trauma to adverse LBP events. For each 6-month study interval, the risk of developing a serious LBP episode was 2.1% unassociated with minor trauma and 2.4% following minor trauma (P = 0.59). Neither the frequency of minor trauma events nor the reported severity of the event correlated with adverse outcomes. Subjects with advanced structural findings were not more likely to become symptomatic with minor trauma events than with spontaneously evolving LBP episodes. Follow-up magnetic resonance imaging evaluating new serious LBP illness rarely revealed new clinically significant findings. Age and sex-adjusted prediction models, including abnormal psychometric testing, smoking, and compensation issues, accurately identified 80% of serious LBP events and 93% of LBP disability events. CONCLUSIONS: In this study cohort, minor trauma does not appear to increase the risk of serious LBP episodes or disability. The vast majority of incident-adverse LBP events may be predicted not by structural findings or minor trauma but by a small set of demographic and behavioral variables. PMID- 17139226 TI - Predictors of pain and function in persons with spinal stenosis, low back pain, and no back pain. AB - STUDY DESIGN: Longitudinal masked, double-controlled cohort study. OBJECTIVES: To determine prognosis and predictors of function and pain in persons with spinal stenosis. SUMMARY OF BACKGROUND DATA: The clinical syndrome of spinal stenosis is common and disabling, but not clearly related to anatomic measures. Prognosis not well studied. METHODS: Persons 55 to 80 years of age with and without stenosis on preliminary review of magnetic resonance imaging (MRI), and asymptomatic volunteers underwent screening, questionnaires, physical examination, ambulation testing, masked electromyogram (EMG), and masked MRI scans; these were repeated at >18 months. RESULTS: Twenty-three asymptomatic, 28 back pain, and 32 clinically diagnosed stenosis subjects underwent follow-up. Although initial and follow-up diagnosis tended to agree (kappa = 0.394, P < 001), there were substantial shifts between the three groups. Among persons with clinically diagnosed stenosis, every measure trended for improvement, including significant changes in pain, ambulation, and EMG. Ambulation velocity and Pain Disability Index at follow-up were predicted by initial disability measures. Pain was predicted by initial sleep difficulty but not initial pain. EMG and MRI did not predict function or pain. CONCLUSION: Clinically recognized spinal stenosis is fluctuating and largely improving, and in continuum with back pain and no symptoms. Since anatomic and neurologic deficits do not predict future function, they should not be weighed heavily in surgical risk-benefit discussions. PMID- 17139227 TI - Influence of X Stop on neural foramina and spinal canal area in spinal stenosis. AB - STUDY DESIGN: Measurements of cross sections of exit foramen and spinal canal were performed before and after placement of X Stop in physiologic postures using positional MR scanner at the stenosed level in patients with lumbar spinal stenosis. OBJECTIVE: To quantify the effect of the implant in vivo on the lumbar spine at the instrumented levels in various postures. SUMMARY OF BACKGROUND DATA: Dimensions of the spinal canal and neural foramen decrease from flexion to extension. Symptoms of spinal stenosis occur typically in standing or extension. The X Stop device is designed to distract the posterior elements of the stenotic segment and place it in flexion to treat neurogenic claudication. We think that the device will improve the dimension of the canal in standing and extension. METHODS: Twenty-six patients with lumbar spine stenosis underwent a one- or two level X Stop procedure. All had preoperative and postoperative positional MRI in standing, supine, and sitting flexion and extension. Measurements were carried out on the images acquired. RESULTS: Significant increase in the dimensions of the neural foramen and canal area were demonstrated after surgery. CONCLUSIONS: The X Stop device improves the degree of central and foraminal stenosis in vivo. PMID- 17139228 TI - Changes in vertebral wedging rate between supine and standing position and its association with back pain: a prospective study in patients with osteoporotic vertebral compression fractures. AB - STUDY DESIGN: Prospective consecutive series. OBJECTIVE: To analyze supine and standing radiographs and the association of back pain using subjective pain criteria. SUMMARY OF BACKGROUND DATA: It has been considered that there is little correlation between the degree of collapse of the vertebral body and the level of pain. In previous studies, however, measurements have only been based on supine radiographs. Although there were 2 authors who reported the results of supine lateral and standing lateral radiographs in patients with thoracolumbar vertebral fractures, as far as we know, there has not been any detailed report concerning the correlation between radiologic findings using supine and standing lateral radiographs and back pain. METHODS: We examined 100 consecutively treated patients, prospectively. Back pain and the supine and standing radiographs were assessed 1 month after injury. Changes in vertebral wedging rate (WR) from supine to standing position (Delta WR) was reported by the following equation: Delta WR = WR(standing)-WR(supine). RESULTS: The median age of the cohort was 75 years (range, 60-89 years). The median VAS of back pain at supine position, at standing position, and when standing erect was 13, 33, and 41, respectively. The median wedging rate on the supine and standing radiographs were 28% and 37%, respectively (P < 0.001). There was a significant correlation between Delta WR and back pain when standing erect (r = 0.79, P < 0.001). CONCLUSION: Changes in vertebral wedging rate between supine and standing position and its association with back pain may give a clue to the pathogenesis of pain from osteoporotic thoracolumbar vertebral compression fractures. PMID- 17139229 TI - Is obesity overrated as a "risk factor" for poor outcomes in chronic occupational spinal disorders? AB - STUDY DESIGN: Prospective cohort study. OBJECTIVE: To investigate whether obesity is a risk factor for poor socioeconomic outcomes in patients with chronic occupational spinal disorders beginning an interdisciplinary functional restoration program. SUMMARY OF BACKGROUND DATA: Obesity is known as a risk factor for a multitude of physical and mental health disorders. A classification system can define obesity based on body mass index (BMI). Limited evidence is available on the effect of obesity on rehabilitation from chronic occupational spinal disorders. METHODS: A large cohort of chronic occupational spinal disorder patients with extended disability (average 16 months) following work-related injuries (n = 2483) were divided into 5 groups based on BMI. Socioeconomic outcomes were identified through a 1-year post-rehabilitation structured telephone interview. Patients were also assessed before and after intervention with validated questionnaires measuring pain intensity, functional deficits, and depression. The interdisciplinary functional restoration program was individualized for length of disability and severity of physical and psychological deficits. RESULTS: The prevalence of obesity in this population was high. Only 22% of patients were "normal" (BMI <25), while 43% were obese (BMI >30). There was a significant linear trend for males to be progressively less represented as obesity increased. There was also a significant trend for hypertension to be associated with greater levels of obesity. However, there were no significant differences in site of injury, disability duration, or attorney representation among the groups. Most importantly, there were no statistically significant differences in all but 1 of the socioeconomic outcomes among any of the obese groups and the normal weight patients. There was a trend toward higher pretreatment and posttreatment functional assessment scores (i.e., greater perception of disability), and a linear trend for higher pretreatment depression ratings across BMI groups, although they all improved at comparable rates after treatment. CONCLUSIONS: Obesity is more prevalent and severe in chronic occupational spinal disorders when entering a functional restoration program as the final step in a prolonged treatment regimen. We conclude that obesity, even gross obesity, has no significant impact on outcomes of an interdisciplinary functional restoration program for patients disabled an average of 16 months. PMID- 17139230 TI - MMPI disability profile: the least known, most useful screen for psychopathology in chronic occupational spinal disorders. AB - STUDY DESIGN: Prospective study on predicting psychopathology in chronic occupational spinal disorders (COSDs). OBJECTIVE: To assess prevalence of specific profiles on the Minnesota Multiphasic Personality Inventory (MMPI) and their ability to predict psychopathology in a COSD cohort. SUMMARY OF BACKGROUND DATA: In the relatively small number of COSDs that develop chronic pain and disability, the MMPI-2 has been an important part of the psychosocial assessment. Certain profiles have been thought to have a high prevalence in COSD. They have also been widely popularized as predicting certain treatment outcomes, and have often been used to "screen" surgical and rehabilitation candidates. METHOD: Of an initial cohort of 1,489 consecutive COSD patients completing a valid prerehabilitation MMPI-2, 1,185 patients (79.6%) were classifiable into one of four MMPI profile groups. A new Disability Profile (DP) group was identified, which was the most common profile. Patients attended a 5- to 7-week interdisciplinary rehabilitation program. They completed a psychosocial assessment battery, and a Structured Clinical Interview for DSM IV diagnosis (SCID-I and II) was administered as the "gold standard" for defining psychopathology. One year postrehabilitation, a structured clinical interview assessed socioeconomic outcomes. RESULTS: A previously unrecognized MMPI profile, now termed the DP, was found to have a prevalence of 53.2% of the whole group, and 66.9% of those with "classifiable" MMPI profiles in this large population of COSD patients. Only 6.9% of subjects had normal profiles (NP), while only 19.5% had profiles previously thought to occur commonly in this population. NP patients were twice as likely to retain work 1 year after treatment than the 3 abnormal MMPI groups combined. The DP group was 14 times more;1 likely to have an Axis I diagnosis (such as depression or anxiety) than the NP group, and was also almost 5 times more likely have an Axis II personality disorder diagnosis. CONCLUSIONS: The prevalence of commonly cited MMPI profiles, often used for presurgical or chronic pain screening in this population, is relatively small. The prevalence of four or more elevations (DP), however, is large, representing two thirds of patients demonstrating any classifiable MMPI pattern. The DP group showed extremely high levels of associated psychopathology, which raises "red flags" to the surgeon likely to operate on such patients, or the interdisciplinary pain team. Screening COSD patients with the MMPI-2 may be effective in identifying psychopathology, but only if the physician is aware that the DP is commonplace and significant. PMID- 17139231 TI - Cluster analysis as a method for determining size ranges for spinal implants: disc lumbar replacement prosthesis dimensions from magnetic resonance images. AB - STUDY DESIGN: Statistical analysis of clinical radiologic data. OBJECTIVE: To develop an objective method for finding the number of sizes for a lumbar disc replacement. SUMMARY OF BACKGROUND DATA: Cluster analysis is a well-established technique for sorting observations into clusters so that the "similarity level" is maximal if they belong to the same cluster and minimal otherwise. METHODS: Magnetic resonance scans from 69 patients, with no abnormal discs, yielded 206 sagittal and transverse images of 206 discs (levels L3-L4-L5-S1). Anteroposterior and lateral dimensions were measured from vertebral margins on transverse images; disc heights were measured from sagittal images. Hierarchical cluster analysis was performed to determine the number of clusters followed by nonhierarchical (K means) cluster analysis. Discriminant analysis was used to determine how well the clusters could be used to classify an observation. RESULTS: The most successful method of clustering the data involved the following parameters: anteroposterior dimension; lateral dimension (both were the mean of results from the superior and inferior margins of a vertebral body, measured on transverse images); and maximum disc height (from a midsagittal image). These were grouped into 7 clusters so that a discriminant analysis was capable of correctly classifying 97.1% of the observations. The mean and standard deviations for the parameter values in each cluster were determined. CONCLUSIONS: Cluster analysis has been successfully used to find the dimensions of the minimum number of prosthesis sizes required to replace L3-L4 to L5-S1 discs; the range of sizes would enable them to be used at higher lumbar levels in some patients. PMID- 17139233 TI - Predictors of human papillomavirus vaccination acceptability among underserved women. AB - OBJECTIVES: The objectives of this study were to examine underserved women's acceptability of the forthcoming human papillomavirus (HPV) vaccines and to identify correlates of HPV vaccine acceptability. STUDY DESIGN: A sample of primarily low-income minority women (n = 58) recruited from community health clinics completed a semistructured interview assessing health beliefs, vaccination attitudes, health behavior, and HPV vaccination intentions. RESULTS: Personal acceptability of the HPV vaccines was generally high. Moreover, 100% of parents were interested in having their children vaccinated. Correlates of vaccination intentions included health beliefs and attitudes (perceived risk of HPV infection, perceived safety and effectiveness of HPV vaccines, perceived physician encouragement for vaccination) and previous health behavior (HIV testing). Independent predictors of vaccine acceptability were also identified. CONCLUSIONS: The current study highlights key correlates of vaccine acceptability that may inform HPV vaccination campaigns for underserved populations. PMID- 17139234 TI - On-site rapid antenatal syphilis screening with an immunochromatographic strip improves case detection and treatment in rural South African clinics. AB - OBJECTIVES: Congenital syphilis is a significant cause of adverse pregnancy outcomes. In South Africa, rural clinics perform antenatal screening offsite, but unreliable transport and poor client follow up impede effective treatment. We compared 3 syphilis screening strategies at rural clinics: on-site rapid plasma reagin (RPR), on-site treponemal immunochromatographic strip (ICS) test, and the standard practice offsite RPR with Treponema pallidum hemagglutination assay (RPR/TPHA). METHODS: Eight rural clinics performed the on-site RPR and ICS tests and provided immediate treatment. Results were compared with RPR/TPHA at a reference laboratory. Chart reviews at 8 standard practice clinics established diagnosis and treatment rates for offsite RPR/TPHA. FINDINGS: Seventy-nine (6.3%) of 1,250 women screened on-site had active syphilis according to the reference laboratory. The on-site ICS resulted in the highest percentage of pregnant women correctly diagnosed and treated for syphilis (89.4% ICS, 63.9% on-site RPR, 60.8% offsite RPR/TPHA). The on-site RPR had low sensitivity (71.4% for high-titer syphilis). The offsite approach suffered from poor client return rates. One percent of women screened with the ICS may have received penicillin unnecessarily. There were no adverse treatment outcomes. CONCLUSIONS: The on-site ICS test can reduce syphilis-related adverse outcomes of pregnancy through accurate diagnosis and immediate treatment of pregnant women with syphilis. PMID- 17139235 TI - Factors and the sociosexual network associated with a syphilis outbreak in rural North Carolina. AB - OBJECTIVE: An investigation was conducted to determine factors associated with a syphilis outbreak in a rural North Carolina county. STUDY DESIGN: A retrospective chart review was performed on 61 primary (PS), secondary (SS), and early latent (ELS) syphilis case patients reported in Columbus County between January 2001 and February 2002. Sociosexual network analysis was conducted using electronic contact tracing information. RESULTS: We identified 20 PS, 25 SS, and 16 ELS case patients who were predominantly black. Seventy-two percent had reported >or=1 sexual partner with early syphilis, 51% used crack cocaine and/or had sex with a crack-using partner, and 31% exchanged sex for drugs or money. The sexual network exhibited predominantly linear connections between case patients and sexual partners. Adding social connections to the network further demonstrated dense cyclic interactions characteristic of core groups. CONCLUSIONS: The syphilis outbreak in this rural community was associated with crack cocaine and exchange of sex for drugs in a densely interconnected sociosexual network. PMID- 17139236 TI - [Basic evaluation of the new pulse sequence for simultaneous acquisition of T1- and T2-weighted images]. AB - A novel pulse sequence that enables simultaneous acquisition of T1-weighted (T1W) and T2-weighted (T2W) images is presented. In this new technique, the inversion recovery (IR) pulse of conventional fast inversion recovery (Fast IR) is replaced with a pulse train that consists of a fast spin echo (FSE) and 180 (y) +90 (x) for driven inversion (DI). By using a shorter TI and independent k-space ordering, the first part of the sequence provides T2W images and the second part provides T1W images, thereby enabling simultaneous acquisition in a single scan time comparable to that of Fast IR. Signal simulation also was conducted, and this was compared with conventional scanning techniques using normal volunteers. In the human studies, both T1W and T2W images showed the same image quality as conventional images, suggesting the potential for this technique to replace the combination of Fast IR and T2W FSE for scan-time reduction. PMID- 17139237 TI - [Optimization of three-dimensional triple IR fast spoiled gradient recalled acquisition in the steady state (FSPGR) to decrease vascular artifact at 3.0 Tesla]. AB - The purpose of this study was to decrease vascular artifacts caused by the in flow effect in three-dimensional inversion recovery prepared fast spoiled gradient recalled acquisition in the steady state (3D IR FSPGR) at 3.0 Tesla. We developed 3D triple IR (3IR) FSPGR and examined the signal characteristics of the new sequence. We have optimized scan parameters based on simulation, phantom, and in-vivo studies. As a result, optimized parameters (1st TI=600 ms, 3rd TI=500 ms) successfully have produced the vessel signal at more than 40% reduction, while gray-white matter contrast was preserved. Moreover, the reduced artifact was also confirmed by visual inspection of the in-vivo images for which this condition was used. Thus, 3D 3IR FSPGR was a useful sequence for the acquisition of T1-weighted images at 3.0 Tesla. PMID- 17139238 TI - [Status of measures against angiography room infection as determined by questionnaire]. AB - Although the cleanliness of the angiography room and that of the operating room have long been equally attended to, the concept of Standard Precautions (including the basic measures and procedures to prevent infection) of the Centers for Disease Control and Prevention (CDC), 1996, as well as the introduction of transmission-based precautions, have been changing to preventive measures that are based on concrete measures. Therefore, a questionnaire was introduced in order to determine the actual status of countermeasures against infection used in the angiography room. The questionnaire was sent to 530 institutions, and 286 responded, a response rate of 54.0%. Its results significantly revealed the following: 1) unexpectedly low recognition of the need and importance for the CDC preventive measures against infection, 2) a considerable number of institutions continuing to perform the conventional preventive measures, 3) problems with education systems on preventive measures, and 4) handwashing, the most important measure against infection, failing to be adequately carried out noticeably among radiological technologists. PMID- 17139239 TI - [Importance of an initial check-up for diagnostic radiology apparatus management]. AB - The management of apparatus for diagnostic imaging is an important job for radiological technologists. In the last 10 years we have encountered 2227 malfunctions in 3652 days. We detected 163 cases at the initial check-up, which accounted for 7.3% of total cases. By performing an initial check-up, we detected one malfunction every 21 days and were able to keep the effects of mishaps to a minimum, prevent accidents, and obtain stable-quality images. PMID- 17139240 TI - Managing patients with low-risk MDS. AB - Supportive care with blood transfusion and administration of hematopoietic growth factors (eg, erythropoietin, colony-stimulating factors) has been the standard of care for patients with low-risk myelodysplastic syndromes (MDS), a group of disorders characterized by hyperproliferation of the bone marrow and ineffective hematopoiesis. However, the development of new drugs, including lenalidomide, azacitidine, and decitabine, has led to a new era of more effective treatment for MDS. Further, the use of classification and risk stratification has allowed for the identification of individuals who are expected to benefit from some therapies while simultaneously excluding over-treatment and unnecessary toxicity in those who are unlikely to benefit from specific drugs. In this Clinical Roundtable Monograph, the faculty discusses the epidemiology, classification, and risk stratification for MDS, medical and nursing issues associated with supportive care and the new therapies for low-risk MDS, and patient education and other strategies for the optimization of quality of life in patients with low-risk MDS. PMID- 17139241 TI - The role of targeted therapy in the treatment of colorectal cancer. AB - Recent years have brought significant advances in the treatment of metastatic colorectal cancer. Combination regimens with standard chemotherapeutic agents have extended survival to nearly 2 years, and recent studies suggest that chemotherapy-free intervals may be feasible in some patients without compromising survival outcomes. The most significant recent progress has centered on the use of targeted biologic therapies. The first targeted agent to show a significant benefit in metastatic colorectal cancer was bevacizumab. This monoclonal antibody is directed against vascular endothelial growth factor, a molecule known to be involved in the angiogenic process that is central to cancer growth and metastasis. In clinical trials, bevacizumab has improved survival when added to multiple chemotherapy regimens. The second targeted agent to be approved for colorectal cancer is the monoclonal antibody cetuximab, which is directed against the epidermal growth factor receptor, another key mediator of cancer growth. Cetuximab has been shown to increases the efficacy of irinotecan in irinotecan refractory patients, indicating that cetuximab may make tumors more sensitive to chemotherapeutic agents. Bevacizumab and cetuximab continue to be evaluated alone as maintenance therapy and in combination in different settings to determine their optimal use in colorectal cancer. Additional targeted agents are also being developed and are showing promise in clinical trials. PMID- 17139243 TI - Indolent lymphomas: current and emerging treatment approaches. AB - The introduction of targeted therapy in combination with chemotherapy has greatly advanced the treatment of patients with indolent lymphomas in recent years. Most commonly studied in patients with follicular lymphoma, the monoclonal antibody rituximab has not only improved the response rate and progression-free survival for both treatment-naive and relapsed/refractory patients, it is also beginning to demonstrate an improvement in overall survival. These benefits are challenging many of the standard treatment approaches in follicular lymphoma and offer hope that treatment of the disease may soon advance from being palliative to curative. Ongoing research seeks to determine how best to use targeted agents in the setting of indolent lymphoma, either concurrently with chemotherapy or sequentially following the administration of chemotherapy. Various studies aim to define the utility of rituximab as a long-term maintenance treatment in several follicular lymphoma settings, as well as its benefit in autologous stem cell transplantation. In addition to these immunotherapy studies, the use of radioimmunotherapy is also being explored for its efficacy as a frontline and consolidation treatment approach. As additional molecular and targeted therapeutics advance through clinical trials, future treatment strategies for indolent lymphomas will likely involve the addition of these newer therapeutics to immunochemotherapy based on the individual risk profile of a patient. PMID- 17139244 TI - The role of angiogenesis inhibition in the treatment of breast cancer. AB - In recent years, antiangiogenic therapy with the monoclonal antibody bevacizumab has demonstrated significant activity in patients with metastatic breast cancer. Bevacizumab is targeted against vascular endothelial growth factor (VEGF), a primary mediator of angiogenesis. Research is ongoing to define the mechanism of action of anti-VEGF treatment in order to predict who will respond to treatment and to monitor responses to treatment at the molecular level. The initial randomized phase III trial of bevacizumab evaluated capecitabine with bevacizumab versus capecitabine alone in patients with heavily pretreated metastatic breast cancer. The addition of bevacizumab to capecitabine did not improve progression free survival in these patients. However, in the subsequent Eastern Cooperative Oncology Group 2100 trial of patients with previously untreated metastatic breast cancer, bevacizumab combined with paclitaxel doubled progression-free survival compared to paclitaxel alone. Based on these encouraging findings, current studies are evaluating bevacizumab in the adjuvant setting. The oral tyrosine kinase inhibitor sunitinib has shown activity in metastatic breast cancer, and additional agents are being investigated. Combination therapy consisting of antiangiogenic agents with chemotherapy, hormone therapy, or other agents is also being evaluated in hopes of improving treatment options for these patients. PMID- 17139245 TI - RecFOR proteins are essential for Pol V-mediated translesion synthesis and mutagenesis. AB - When the replication fork moves through the template DNA containing lesions, daughter-strand gaps are formed opposite lesion sites. These gaps are subsequently filled-in either by translesion synthesis (TLS) or by homologous recombination. RecA filaments formed within these gaps are key intermediates for both of the gap-filling pathways. For instance, Pol V, the major lesion bypass polymerase in Escherichia coli, requires a functional interaction with the tip of the RecA filament. Here, we show that all three recombination mediator proteins RecFOR are needed to build a functionally competent RecA filament that supports efficient Pol V-mediated TLS in the presence of ssDNA-binding protein (SSB). A positive contribution of RecF protein to Pol V lesion bypass is demonstrated. When Pol III and Pol V are both present, Pol III imparts a negative effect on Pol V-mediated lesion bypass that is counteracted by the combined action of RecFOR and SSB. Mutations in recF, recO or recR gene abolish induced mutagenesis in E. coli. PMID- 17139246 TI - ATAB2 is a novel factor in the signalling pathway of light-controlled synthesis of photosystem proteins. AB - Plastid translational control depends to a large extent on the light conditions, and is presumably mediated by nucleus-encoded proteins acting on organelle gene expression. However, the molecular mechanisms of light signalling involved in translation are still poorly understood. We investigated the role of the Arabidopsis ortholog of Tab2, a nuclear gene specifically required for translation of the PsaB photosystem I subunit in the unicellular alga Chlamydomonas. Inactivation of ATAB2 strongly affects Arabidopsis development and thylakoid membrane biogenesis and leads to an albino phenotype. Moreover the rate of synthesis of the photosystem reaction center subunits is decreased and the association of their mRNAs with polysomes is affected. ATAB2 is a chloroplast A/U rich RNA-binding protein that presumably functions as an activator of translation with at least two targets, one for each photosystem. During early seedling development, ATAB2 blue-light induction is lowered in photoreceptor mutants, notably in those lacking cryptochromes. Considering its role in protein synthesis and its photoreceptor-mediated expression, ATAB2 represents a novel factor in the signalling pathway of light-controlled translation of photosystem proteins during early plant development. PMID- 17139247 TI - GLUT4 is internalized by a cholesterol-dependent nystatin-sensitive mechanism inhibited by insulin. AB - Insulin slows GLUT4 internalization by an unknown mechanism. Here we show that in unstimulated adipocytes, GLUT4 is internalized by two mechanisms. Approximately 80% of GLUT4 is internalized by a mechanism that is sensitive to the cholesterol aggregating drug nystatin, and is independent of AP-2 clathrin adaptor and two putative GLUT4 endocytic motifs. The remaining GLUT4 is internalized by an AP-2 dependent, nystatin-resistant pathway that requires the FQQI GLUT4 motif. Insulin inhibits GLUT4 uptake by the nystatin-sensitive pathway and, consequently, GLUT4 is internalized by the AP-2-dependent pathway in stimulated adipocytes. The phenylalanine-based FQQI GLUT4 motif promotes AP-2-dependent internalization less rapidly than a tyrosine-based motif, the classic form of aromatic-based motifs. Thus, both a change in the predominant endocytosis pathway and the specific use of a suboptimal internalization motif contribute to the slowing of GLUT4 internalization in insulin-stimulated adipocytes. Insulin also inhibits the uptake of cholera-toxin B, indicating that insulin broadly regulates cholesterol dependent uptake mechanisms rather than specially targeting GLUT4. Our work thus identifies cholesterol-dependent uptake as a novel target of insulin action in adipocytes. PMID- 17139248 TI - Drosophila homologs of mammalian TNF/TNFR-related molecules regulate segregation of Miranda/Prospero in neuroblasts. AB - During neuroblast (NB) divisions, cell fate determinants Prospero (Pros) and Numb, together with their adaptor proteins Miranda (Mira) and Partner of Numb, localize to the basal cell cortex at metaphase and segregate exclusively to the future ganglion mother cells (GMCs) at telophase. In inscuteable mutant NBs, these basal proteins are mislocalized during metaphase. However, during anaphase/telophase, these mutant NBs can partially correct these earlier localization defects and redistribute cell fate determinants as crescents to the region where the future GMC "buds" off. This compensatory mechanism has been referred to as "telophase rescue". We demonstrate that the Drosophila homolog of the mammalian tumor-necrosis factor (TNF) receptor-associated factor (DTRAF1) and Eiger (Egr), the homolog of the mammalian TNF, are required for telophase rescue of Mira/Pros. DTRAF1 localizes as an apical crescent in metaphase NBs and this apical localization requires Bazooka (Baz) and Egr. The Mira/Pros telophase rescue seen in inscuteable mutant NBs requires DTRAF1. Our data suggest that DTRAF1 binds to Baz and acts downstream of Egr in the Mira/Pros telophase rescue pathway. PMID- 17139249 TI - GSK-3beta-regulated interaction of BICD with dynein is involved in microtubule anchorage at centrosome. AB - Microtubule arrays direct intracellular organization and define cellular polarity. Here, we show a novel function of glycogen synthase kinase-3beta (GSK 3beta) in the organization of microtubule arrays through the interaction with Bicaudal-D (BICD). BICD is known to form a complex with dynein-dynactin and to function in the intracellular vesicle trafficking. Our data revealed that GSK 3beta is required for the binding of BICD to dynein but not to dynactin. Knockdown of GSK-3beta or BICD reduced centrosomally focused microtubules and induced the mislocalization of centrosomal proteins. The unfocused microtubules in GSK-3beta knockdown cells were rescued by the expression of the dynein intermediate chain-BICD fusion protein. Microtubule regrowth assays showed that GSK-3beta and BICD are required for the anchoring of microtubules to the centrosome. These results imply that GSK-3beta may function in transporting centrosomal proteins to the centrosome by stabilizing the BICD1 and dynein complex, resulting in the regulation of a focused microtubule organization. PMID- 17139250 TI - The Galpha12-RGS RhoGEF-RhoA signalling pathway regulates neurotransmitter release in C. elegans. AB - In Caenorhabditis elegans adults, the single Rho GTPase orthologue, RHO-1, stimulates neurotransmitter release at synapses. We show that one of the pathways acting upstream of RHO-1 in acetylcholine (ACh)-releasing motor neurons depends on Galpha12 (GPA-12), which acts via the single C. elegans RGS RhoGEF (RHGF-1). Activated GPA-12 has the same effect as activated RHO-1, inducing the accumulation of diacylglycerol and the neuromodulator UNC-13 at release sites, and increased ACh release. We showed previously that RHO-1 stimulates ACh release by two separate pathways-one that requires UNC-13 and a second that does not. We show here that a non-DAG-binding-UNC-13 mutant that partially blocks increased ACh release by activated RHO-1 completely blocks increased ACh release by activated GPA-12. Thus, the upstream GPA-12/RHGF-1 pathway stimulates only a subset of RHO-1 downstream effectors, suggesting that either the RHO-1 effectors require different levels of activated RHO-1 for activation or there are two distinct pools of RHO-1 within C. elegans neurons. PMID- 17139251 TI - p38 kinase regulates epidermal growth factor receptor downregulation and cellular migration. AB - Internalization and proteolytic degradation of epidermal growth factor (EGF) receptor (R) following ligand binding is an important mechanism for regulating EGF-stimulated signals. Using pharmacological and RNA interference inhibition of p38 mitogen-activated protein kinase, we show that p38 is required for efficient EGF-induced EGFR destruction but not internalization. In the absence of p38 activity, EGF fails to stimulate the ubiquitin ligase Cbl or ubiquitinylation of EGFR, and internalized EGFR accumulates in intracellular vesicles containing caveolin-1. These effects are accompanied by loss of EGFR phosphorylation on Y1045, a phosphorylation site required for Cbl activation. Furthermore, similar to cells treated with p38 inhibitors, intestinal epithelial cells expressing Y1045F EGFR mutants show increased proliferation but not migration in response to EGF, thus uncoupling these biological responses. Together these data position p38 as a modulator of ligand-stimulated EGFR processing and demonstrate that this processing has a profound impact on the cellular outcome of EGFR signaling. PMID- 17139253 TI - NoRC-dependent nucleosome positioning silences rRNA genes. AB - Previous studies have established that the Snf2h-containing chromatin remodeling complex NoRC mediates epigenetic silencing of a subset of rRNA genes (rDNA) by recruiting enzymatic activities that modify histones and methylate DNA. Here we have analyzed nucleosome positions at the murine rDNA promoter and show that active and silent rDNA copies are characterized not only by specific epigenetic marks but also by differently positioned nucleosomes. At active genes the promoter-bound nucleosome covers nucleotides from -157 to -2, whereas at silent genes the nucleosome is positioned 25 nucleotides further downstream. We provide evidence that NoRC is the molecular machine that shifts the promoter-bound nucleosome downstream of the transcription start site into a translational position that is unfavorable for transcription complex formation. PMID- 17139252 TI - c-myc as a mediator of accelerated apoptosis and involution in mammary glands lacking Socs3. AB - Suppressor of cytokine signalling (SOCS) proteins are critical attenuators of cytokine-mediated signalling in diverse tissues. To determine the importance of Socs3 in mammary development, we generated mice in which Socs3 was deleted in mammary epithelial cells. No overt phenotype was evident during pregnancy and lactation, indicating that Socs3 is not a key physiological regulator of prolactin signalling. However, Socs3-deficient mammary glands exhibited a profound increase in epithelial apoptosis and tissue remodelling, resulting in precocious involution. This phenotype was accompanied by augmented Stat3 activation and a marked increase in the level of c-myc. Moreover, induction of c myc before weaning using an inducible transgenic model recapitulated the Socs3 phenotype, and elevated expression of likely c-myc target genes, E2F-1, Bax and p53, was observed. Our data establish Socs3 as a critical attenuator of pro apoptotic pathways that act in the developing mammary gland and provide evidence that c-myc regulates apoptosis during involution. PMID- 17139254 TI - Repression of ADH1 and ADH3 during zinc deficiency by Zap1-induced intergenic RNA transcripts. AB - The transcriptional activator Zap1 induces target gene expression in response to zinc deficiency. We demonstrate that during zinc starvation, Zap1 is required for the repression of ADH1 expression. ADH1 encodes the major zinc-dependent alcohol dehydrogenase that is utilized during fermentation. During zinc starvation, Zap1 binds upstream of the activator Rap1 and induces an intergenic RNA transcript, ZRR1. ZRR1 expression leads to the transient displacement of Rap1 from the ADH1 promoter resulting in ADH1 repression. Using a microarray-based approach, we screened for additional genes repressed by Zap1 intergenic transcripts. We found that ADH3, the major mitochondrial alcohol dehydrogenase, is regulated in a manner similar to ADH1. Thus, during zinc deficiency, Zap1 mediates the repression of two of the most abundant zinc-requiring enzymes. PMID- 17139255 TI - Structure of the origin-binding domain of simian virus 40 large T antigen bound to DNA. AB - The large T antigen (T-ag) protein binds to and activates DNA replication from the origin of DNA replication (ori) in simian virus 40 (SV40). Here, we determined the crystal structures of the T-ag origin-binding domain (OBD) in apo form, and bound to either a 17 bp palindrome (sites 1 and 3) or a 23 bp ori DNA palindrome comprising all four GAGGC binding sites for OBD. The T-ag OBDs were shown to interact with the DNA through a loop comprising Ser147-Thr155 (A1 loop), a combination of a DNA-binding helix and loop (His203-Asn210), and Asn227. The A1 loop traveled back-and-forth along the major groove and accounted for most of the sequence-determining contacts with the DNA. Unexpectedly, in both T-ag-DNA structures, the T-ag OBDs bound DNA independently and did not make direct protein protein contacts. The T-ag OBD was also captured bound to a non-consensus site ATGGC even in the presence of its canonical site GAGGC. Our observations taken together with the known biochemical and structural features of the T-ag-origin interaction suggest a model for origin unwinding. PMID- 17139256 TI - Multiple mechanisms are involved in regulating the expression of the developmental timing regulator lin-28 in Caenorhabditis elegans. AB - The timing of postembryonic developmental programs in Caenorhabditis elegans is regulated by a set of so-called heterochronic genes, including lin-28 that specifies second larval programs. lin-66 mutations described herein cause delays in vulval and seam cell differentiation, indicating a role for lin-66 in timing regulation. A mutation in daf-12/nuclear receptor or alg-1/argonaute dramatically enhances the retarded phenotypes of the lin-66 mutants, and these phenotypes are suppressed by a lin-28 null allele. We further show that the LIN-28 protein level is upregulated in the lin-66 mutants and that this regulation is mediated by the 3'UTR of lin-28. We have also identified a potential daf-12-response element within lin-28 3'UTR and show that two microRNA (miRNA) (lin-4 and let-7)-binding sites mediate redundant inhibitory activities that are likely lin-66-independent. Quantitative PCR data suggest that the lin-28 mRNA level is affected by lin-14 and miRNA regulation, but not by daf-12 and lin-66 regulation. These results suggest that lin-28 expression is regulated by multiple independent mechanisms including LIN-14-mediated upregulation of mRNA level, miRNAs-mediated RNA degradation, LIN-66-mediated translational inhibition and DAF-12-involved translation promotion. PMID- 17139257 TI - hRpn13/ADRM1/GP110 is a novel proteasome subunit that binds the deubiquitinating enzyme, UCH37. AB - The 26S proteasome catalyzes the degradation of most proteins in mammalian cells. To better define its composition and associated regulatory proteins, we developed affinity methods to rapidly purify 26S proteasomes from mammalian cells. By this approach, we discovered a novel 46-kDa (407 residues) subunit of its 19S regulatory complex (previously termed ADRM1 or GP110). As its N-terminal half can be incorporated into the 26S proteasome and is homologous to Rpn13, a 156-residue subunit of the 19S complex in budding yeast, we renamed it human Rpn13 (hRpn13). The C-terminal half of hRpn13 binds directly to the proteasome-associated deubiquitinating enzyme, UCH37, and enhances its isopeptidase activity. Knockdown of hRpn13 in 293T cells increases the cellular levels of ubiquitin conjugates and decreases the degradation of short-lived proteins. Surprisingly, an overproduction of hRpn13 also reduced their degradation. Furthermore, transfection of the C-terminal half of hRpn13 slows proteolysis and induces cell death, probably by acting as a dominant-negative form. Thus in human 26S proteasomes, hRpn13 appears to be important for the binding of UCH37 to the 19S complex and for efficient proteolysis. PMID- 17139258 TI - Asymmetric conformational changes in a GPCR dimer controlled by G-proteins. AB - G-protein-coupled receptors (GPCRs) are key players in cell communication. Although long considered as monomeric, it now appears that these heptahelical proteins can form homo- or heterodimers. Here, we analyzed the conformational changes in each subunit of a receptor dimer resulting from agonist binding to either one or both subunits by measuring the fluorescent properties of a leukotriene B(4) receptor dimer with a single 5-hydroxytryptophan-labeled protomer. We show that a receptor dimer with only a single agonist-occupied subunit can trigger G-protein activation. We also show that the two subunits of the receptor dimer in the G-protein-coupled state differ in their conformation, even when both are liganded by the agonist. No such asymmetric conformational changes are observed in the absence of G-protein, indicating that the interaction of the G-protein with the receptor dimer brings specific constraints that prevent a symmetric functioning of this dimer. These data open new options for the differential signaling properties of GPCR dimers. PMID- 17139259 TI - DNA segregation by the bacterial actin AlfA during Bacillus subtilis growth and development. AB - We here identify a protein (AlfA; actin like filament) that defines a new family of actins that are only distantly related to MreB and ParM. AlfA is required for segregation of Bacillus subtilis plasmid pBET131 (a mini pLS32-derivative) during growth and sporulation. A 3-kb DNA fragment encoding alfA and a downstream gene (alfB) is necessary and sufficient for plasmid stability. AlfA-GFP assembles dynamic cytoskeletal filaments that rapidly turn over (t(1/2)< approximately 45 s) in fluorescence recovery after photobleaching experiments. A point mutation (alfA D168A) that completely inhibits AlfA subunit exchange in vivo is strongly defective for plasmid segregation, demonstrating that dynamic polymerization of AlfA is necessary for function. During sporulation, plasmid segregation occurs before septation and independently of the DNA translocase SpoIIIE and the chromosomal Par proteins Soj and Spo0J. The absence of the RacA chromosome anchoring protein reduces the efficiency of plasmid segregation (by about two fold), suggesting that it might contribute to anchoring the plasmid at the pole during sporulation. Our results suggest that the dynamic polymerization of AlfA mediates plasmid separation during both growth and sporulation. PMID- 17139260 TI - X-ray structure of the membrane-bound cytochrome c quinol dehydrogenase NrfH reveals novel haem coordination. AB - Oxidation of membrane-bound quinol molecules is a central step in the respiratory electron transport chains used by biological cells to generate ATP by oxidative phosphorylation. A novel family of cytochrome c quinol dehydrogenases that play an important role in bacterial respiratory chains was recognised in recent years. Here, we describe the first structure of a cytochrome from this family, NrfH from Desulfovibrio vulgaris, which forms a stable complex with its electron partner, the cytochrome c nitrite reductase NrfA. One NrfH molecule interacts with one NrfA dimer in an asymmetrical manner, forming a large membrane-bound complex with an overall alpha(4)beta(2) quaternary arrangement. The menaquinol-interacting NrfH haem is pentacoordinated, bound by a methionine from the CXXCHXM sequence, with an aspartate residue occupying the distal position. The NrfH haem that transfers electrons to NrfA has a lysine residue from the closest NrfA molecule as distal ligand. A likely menaquinol binding site, containing several conserved and essential residues, is identified. PMID- 17139261 TI - p53 mediates the negative regulation of MDM2 by orphan receptor TR3. AB - MDM2 is an oncoprotein whose transforming potential is activated by overexpression. The expression level of MDM2 is negatively regulated by orphan receptor TR3 that mainly acts as a transcriptional factor to regulate gene expression. However, the underlying mechanism is largely unclear. Here, we present the first evidence that inhibition of TR3 on MDM2 is mediated by p53. We found that TR3 directly interacts with p53 but not MDM2, and such interaction is critical for TR3 to inhibit MDM2 expression. TR3 downregulates p53 transcriptional activity by blocking its acetylation, leading to a decrease on the transcription level of MDM2. Furthermore, TR3 binding to p53 obstructs its ubiquitination and degradation induced by MDM2, resulting in the MDM2 ubiquitination and degradation. In addition, TR3 could enhance p53-mediated apoptosis induced by UV irradiation. Taken together, our findings demonstrate that p53 mediates the suppression of TR3 on MDM2 at both transcriptional and post transcriptional level and suggest TR3 as a potential target to develop new anticancer agents that restrict MDM2-induced tumor progression. PMID- 17139262 TI - Soluble FAS ligand: a discriminating feature between drug-induced skin eruptions and viral exanthemas. AB - The clinical spectrum of cutaneous eruptions comprises benign variants like maculopapular rashes (MPRs) and potentially life-threatening events such as toxic epidermal necrolysis (TEN). Apoptosis of keratinocytes is a common histopathological feature of all these drug eruptions. As in skin lesions of TEN and Stevens-Johnson syndrome patients, apoptosis of keratinocytes is often accompanied by an only sparse cellular infiltrate, a soluble fatty acid synthetase ligand (sFASL)-mediated mechanism of keratinocyte cell death is postulated. In MPR patients, evidence for the occurrence of a similar process could not be established so far. We therefore examined sera and lesional skin sections from patients with clinical variants of drug eruptions for FASL expression using a sandwich ELISA and immunohistochemistry, respectively. As controls, healthy persons and patients with other inflammatory skin diseases such as viral exanthema were analyzed. Elevated levels of FASL were detected not only in TEN patients but also in sera and lesional skin of patients with MPR. In contrast, sFASL was repeatedly negative in all viral exanthemas and healthy controls tested. Thus, determination of sFASL serum concentration may represent a discriminating tool between drug rashes and viral exanthemas. PMID- 17139263 TI - Accumulation of elafin in actinic elastosis of sun-damaged skin: elafin binds to elastin and prevents elastolytic degradation. AB - Elafin has a primary structure with two functional domains; a transglutaminase substrate domain at the N-terminus and a protease inhibitor domain at the C terminus. Elafin expression has so far been reported only for epithelial tissues. Accumulation of elafin was immunohistochemically detected in the actinic elastosis of sun-damaged skin. Exposure of normal skin to UVA induced elafin expression that colocalized with elastic fibers. Incubation of synthetic transglutaminase substrate domain of elafin and elastin molecules in the presence of tissue transglutaminase in vitro resulted in the formation of a higher molecular complex on SDS-PAGE. Elafin expression was not detected in normal cultured skin fibroblasts, but was induced by UVA irradiation at both messenger RNA and protein levels. When radiolabeled insoluble elastin was incubated with recombinant full-length elafin and tissue transglutaminase, insoluble elastin became more resistant to neutrophil elastase digestion. These results indicate that (1) dermal fibroblasts potentially express elafin on UV irradiation, (2) UV mediated elafin interacts with elastin, and (3) the elafin-elastin complex protects elastic fibers from elastolytic degradation, leading to the accumulation of elastic fibers in the actinic elastosis of sun-damaged skin. The transglutaminase substrate moiety of elafin plays an important role in anchoring elafin at its proper sites of action during UV-induced aging processes. PMID- 17139264 TI - Epigallocatechin gallate's protective effect against MMP7 in recessive dystrophic epidermolysis bullosa patients. AB - The analysis of phenotype-genotype correlations of patients suffering from recessive dystrophic epidermolysis bullosa (RDEB) evidenced intrafamilial and interfamilial phenotype variability occurring for the same mutation of COL7A1; this underscores the role of other genetics environmental factors in the expressivity of the disease. In this work, we checked whether matrilysin 1 (matrix metalloproteinase (MMP)7) could take part in the epidermal detachment in RDEB. Furthermore, we investigated epigallocatechin 3 gallate (EGCG) to determine whether it could inhibit matrilysin activities on collagen type VII and fibrillin 1 known to be associated with the dermo-epidermal junction. In this work, matrilysin 1 was detected in affected and unaffected skins of the three RDEB patients; furthermore, MMP7 was shown to degrade ex vivo on healthy normal skin collagen VII and fibrillin 1. Thus, we suspect that MMP7 could take an active part in the epidermal detachment occurring during RDEB. We evidenced that EGCG in in vitro as well as in ex vivo experiments was a good inhibitor of MMP7 and developed a good protection of collagen type VII and fibrillin 1 susceptible of being degraded by MMP7. We therefore propose that EGCG could be used beneficially in patients suffering from RDEB. PMID- 17139265 TI - Upregulation of IL-6, IL-8, CXCL1, and CXCL2 dominates gene expression in human fibroblast cells exposed to Loxosceles reclusa sphingomyelinase D: insights into spider venom dermonecrosis. PMID- 17139266 TI - Dermatopharmacokinetic prediction of topical drug bioavailability in vivo. AB - The overall goal of this study was to explore the potential of using stratum corneum (SC) tape-stripping, post-application of a topical drug formulation, to derive dermatopharmacokinetic parameters describing the rate and extent of delivery into the skin. Ibuprofen was administered in 75:25 v/v propylene glycol water to the ventral forearms of human volunteers for periods ranging between 15 and 180 minutes. Subsequently, SC was tape-stripped, quantified gravimetrically, and extracted for drug analysis. Together with concomitant transepidermal water loss measurements, SC concentration-depth profiles of the drug were reproducibly determined and fitted mathematically. The SC-vehicle partition coefficient (K) and a first-order rate constant related to ibuprofen diffusivity in the membrane (D/L2, where L=SC thickness) were derived from data-fitting and characterized the extent and rate of drug absorption across the skin. Integration of the concentration profiles yielded the total drug amount in the SC at the end of the application period. Using K and D/L2 obtained from the 30-minute exposure, it was possible to predict ibuprofen uptake as a function of time into the SC. Prediction and experiment agreed satisfactorily suggesting that objective and quantitative information, with which to characterize topical drug bioavailability, can be obtained from this approach. PMID- 17139267 TI - Novel homozygous frameshift mutation of EVER1 gene in an epidermodysplasia verruciformis patient. AB - Epidermodysplasia verruciformis (EV) is a rare genetic skin disease with an autosomal recessive trait, and the patients have susceptibility to a specific group of human papillomavirus genotypes. Recently germline mutations in EVER1/2 genes have been detected in EV patients with different ethnic origins. In this study, we have applied PCR, single-stranded conformational polymorphism analysis, and sequencing as well as restriction fragment length polymorphism analysis for identifying potential mutation(s) of EVER genes in an EV patient and in the parents of Pakistani origin. A novel homozygous frameshift mutation (T base deletion at nucleotide position 968 of DNA) has been detected in the EVER1 gene of the patient. The parents carried this mutated allele in a heterozygous form. This is the third report on the presence of EVER1 mutations in an EV patient, and this result supports better understanding, diagnosis, and genetic counseling of EV patients. PMID- 17139268 TI - Novel mutations in ALOX12B in patients with autosomal recessive congenital ichthyosis and evidence for genetic heterogeneity on chromosome 17p13. AB - We report clinical and molecular findings in 20 patients from 11 families with autosomal recessive congenital ichthyosis (ARCI) linked to chromosome 17p13, and attributed to mutations in the ALOX gene cluster, which includes three lipoxygenase genes, ALOXE3, ALOX12B, and ALOX15B. We identified six novel missense mutations and one novel deletion leading to a premature stop codon in ALOX12B in only six out of the 11 families which led us to investigate a possible implication of ALOX15B. Mutation analysis of this gene, as well as ALOXE3, which is known to be mutated in some cases of ARCI, failed to reveal causative mutations in the five remaining ARCI families, indicating that other genes on chromosome 17p13 may be involved in this disease. However, by adding new variants to the repertoire of ALOX12B mutations in non-bullous congenital ichthyosiform erythroderma, our data contribute to an enlargement of the spectrum of mutations for the development of efficient molecular genetic tests for analysis of at risk individuals whose carrier status is unknown. PMID- 17139269 TI - Oligonucleotides suppress IL-8 in skin keratinocytes in vitro and offer anti inflammatory properties in vivo. AB - DNA codes for genetic information. Furthermore, recent findings suggest that DNA offers additional function, particularly in the recognition of microorganisms. In this study, we investigated two classes of oligodeoxynucleotides (ODN) in skin keratinocytes; namely, an ODN comprising two cytidine-phosphate-guanosine (CpG) motifs (CpG-1-phosphorothioate (PTO)) and a poly-cytidine (Non-CpG-5-PTO) as control. Both fluorescence-tagged ODN were rapidly taken up by cells and accumulated already after 5 minutes in perinuclear compartments. In order to test whether ODN convey immunological effects in keratinocytes, secretion of IL-8 was measured. Interestingly, both CpG-1-PTO and Non-CpG-5-PTO suppressed basal and tumor necrosis factor alpha-induced IL-8 levels measured in cell culture supernatants. Experiments using deletion mutant revealed a critical length of approximately 16 nucleotides conveying IL-8 suppression. Studies regarding the ODN backbone offered that PTO bondings are critical for significant IL-8 suppression. In order to substantiate the anti-inflammatory response, a contact hypersensitivity mouse model was utilized. Topical application of Non-CpG-5-PTO containing ointments reduced ear thickness in sensitized mice. Taken together, these findings suggest an anti-inflammatory effect of ODN in epithelial cells in vitro and in vivo, indicating that DNA molecules offer distinct biological activities restricted to the physiological compartment applied. This effect seems to be independent from Toll-like receptor 9. PMID- 17139270 TI - Follow-up analysis of PSORS9 in 151 Chinese families confirmed the linkage to 4q31-32 and refined the evidence to the families of early-onset psoriasis. AB - Psoriasis linkage to 4q28-32 (PSORS9) was initially identified by our genome-wide scan in 61 Chinese families and subsequently supported by a meta-analysis of five genome-wide linkage scans of European populations. In this study, we performed a follow-up analysis of PSORS9 using an additional 90 families and improved marker coverage. Joint analysis of all 151 families obtained significant linkage evidence (HLOD=4.53, nonparametric linkage (NPL)=4.03 (P=0.000003)) at the marker interval D4S2997-D4S3033, and the same was obtained for the analysis of the independent new families (HLOD=4.33, NPL=3.15 (P=0.00004)). The linkage evidences from the whole families and the new families exceeded the genome-wide criteria for significant linkage. Furthermore, by performing an ordered subset analysis using mean age at onset as a covariate, we demonstrated that evidence for linkage to PSORS9 is concentrated in the early-onset families and suggested that further study of PSORS9 should focus on early-onset patients. This finding is contradictory to what was found in the Icelandic population and, together with other linkage results, suggests that Chinese and European populations are genetically different for linkage to PSORS9, which may partially explain the influence of ethnic factors on the varying prevalence of psoriasis. PMID- 17139271 TI - Buckle-related complications following surgical repair of retinal dialysis. AB - AIM: To describe buckle-related complications following surgical repair of retinal dialysis. METHODS: A retrospective study of 28 consecutive cryobuckle procedures for retinal detachments secondary to retinal dialysis is reported, with particular attention directed towards postoperative complications relating to the buckle. Stata 8 statistical software and Fisher's exact test were used to analyse the data. RESULTS: Of the 28 cases, anatomic success was achieved with a single procedure in 26 cases (92.9%). Postoperative complications were seen in 20 cases (71.4%), with complications attributable to the buckle noted in 19 (67.9%). Buckle-related complications included exposure (7; 25%), strabismus (5; 17.9%), and infection (3; 10.7%). Surgical removal of the buckle was indicated in 13 cases (46.4%), typically within the first 6 postoperative months. Of these, the retina remained flat following removal of buckle in 12 cases (92.3%), whereas the retina redetached in one case (7.7%). CONCLUSION: Cryotherapy with explant is an effective primary procedure for the surgical repair of retinal detachment secondary to retinal dialysis. However, there is a high rate of postoperative complications relating to the buckle following this surgical approach, although the buckle can be safely removed without compromising the anatomic success of the primary surgery in the vast majority of cases. PMID- 17139272 TI - Deciphering the code: does clinical coding accurately reflect peroperative cataract surgery complication rates? PMID- 17139273 TI - Paradoxical worsening of ocular tuberculosis in HIV patients after antiretroviral therapy. PMID- 17139274 TI - The impact of new regulations on the incidence and severity of ocular injury sustained in hurling. AB - AIM: To prospectively evaluate, and compare, the incidence of hurling-related eye injuries in the South East of Ireland before and after implementation of new rules rendering the use of protective head gear and face masks compulsory for players aged 18 years or under. METHODS: Details relating to patients attending the regional ophthalmic department with injuries sustained during hurling were prospectively recorded between 1 October 2003 and 31 March 2006. RESULTS: Sixty players attended with hurling-related ocular injuries during the study period. Of these, 43 (71.6%) and 17 (28.3%) sustained the ocular injuries during the 15 month period before, and after implementation of the new rules (1 January 2005), respectively. Restricting our analysis to players aged 18 years or under, and, for statistical validity, to the 12-month period immediately before and immediately after implementation of the new rules, a statistically significant reduction in the number of hurling-related injuries was seen (1 January 2004 to 31 December 2004: 11; 1 January 2005 to 31 December 2005: 2; chi(2) test P<0.05) in this age group. A permanent visual deficit was seen in 11 (18.3%) patients. Of these, one (9%) was aged 18 years or under, and this injury was sustained before the new regulations. CONCLUSION: New rules rendering the use of protective eye wear compulsory for players aged 18 years or under have resulted in a significant reduction in the incidence and severity of hurling-related eye injuries in this age group. PMID- 17139276 TI - Sustainable databases. AB - Although the flood of cell biological knowledge rises relentlessly, many databases face an uncertain future. Unless funding for essential bioinformatic resources is set in stone, the next storm may wash away the foundation of future cell biology research. PMID- 17139275 TI - Lower energy levels adequate for effective transcleral diode laser cyclophotocoagulation in Asian eyes with refractory glaucoma. AB - PURPOSE: To study the treatment parameters for diode laser cyclophotocoagulation (DLCP) in Asian Indian eyes using laser energy titrated to clinical response. METHODS: This prospective interventional longitudinal study included 66 eyes of 66 patients with varied aetiology refractory glaucoma, no previous cycloablation, and minimum 1 year follow-up. DLCP was performed using the Oculight Diode laser system IRIS( Medical Instruments Inc., CA, USA). Power used per spot was titrated according to the audible 'pops' indicating tissue microexplosion. The mean laser energy delivered, post-laser intraocular pressure (IOP) reduction, complications, and requirement of re-treatment in various subgroups were analyzed. Differences in energy delivered in each subgroup were assessed by analysis of variance with post hocBonferroni corrections. Linear regression analysis was used to identify possible predictive factors for failure of cyclodiode therapy. RESULTS: The mean total energy delivered per eye was 87.80+/-31.8 J (range 105.4+/-36.8 J in neovascular glaucoma (NVG) to 61.5+/-8.8 J in uveitic glaucoma (P=0.134)). Mean pre treatment IOP was 36.4+/-10.7 mmHg, which reduced to 19.4+/-9.8 mmHg (P<0.001) at 1 week, and 15.6+/-6.6 mmHg at 1 year. At 1 year, 58 of 66 patients had IOP<22.0 mmHg (response rate 87.8%), and six patients had hypotony (success rate 78.8%). The uveitic glaucoma group had 100% success rate. NVG group required maximum re-treatments. CONCLUSIONS: DLCP with a titrated energy protocol needs resulted in lower energy in Asian Indian eyes compared to that reported in literature, and different energy levels are needed for different diseases. 'Standard treatment parameters' for DLCP may be inappropriate for all diseases and all races. PMID- 17139278 TI - DNA replication timing: random thoughts about origin firing. AB - Regions of metazoan genomes replicate at defined times within S phase. This observation suggests that replication origins fire with a defined timing pattern that remains the same from cycle to cycle. However, an alterative model based on the stochastic firing of origins may also explain replication timing. This model assumes varying origin efficiency instead of a strict origin-timing programme. Here, we discuss the evidence for both models. PMID- 17139279 TI - Upgrading the BCL-2 network. PMID- 17139280 TI - eIF4A goes beyond translation. PMID- 17139281 TI - S100 chemokines mediate bookmarking of premetastatic niches. PMID- 17139282 TI - Voltage sparks a GPCR. PMID- 17139283 TI - Shaped to split. PMID- 17139284 TI - How many drug targets are there? AB - For the past decade, the number of molecular targets for approved drugs has been debated. Here, we reconcile apparently contradictory previous reports into a comprehensive survey, and propose a consensus number of current drug targets for all classes of approved therapeutic drugs. One striking feature is the relatively constant historical rate of target innovation (the rate at which drugs against new targets are launched); however, the rate of developing drugs against new families is significantly lower. The recent approval of drugs that target protein kinases highlights two additional trends: an emerging realization of the importance of polypharmacology, and also the power of a gene-family-led approach in generating novel and important therapies. PMID- 17139285 TI - Mining the Wnt pathway for cancer therapeutics. AB - Aberrant activation of the Wnt pathway is implicated in driving the formation of various human cancers, particularly those of the digestive tract. Inhibition of aberrant Wnt pathway activity in cancer cell lines efficiently blocks their growth, highlighting the great potential of therapeutics designed to achieve this in cancer patients. Here we provide an overview of the promise and pitfalls of current drug development strategies striving to inhibit the Wnt pathway and present new opportunities for therapeutic intervention. PMID- 17139286 TI - Antiviral agents active against influenza A viruses. AB - The recent outbreaks of avian influenza A (H5N1) virus, its expanding geographic distribution and its ability to transfer to humans and cause severe infection have raised serious concerns about the measures available to control an avian or human pandemic of influenza A. In anticipation of such a pandemic, several preventive and therapeutic strategies have been proposed, including the stockpiling of antiviral drugs, in particular the neuraminidase inhibitors oseltamivir (Tamiflu; Roche) and zanamivir (Relenza; GlaxoSmithKline). This article reviews agents that have been shown to have activity against influenza A viruses and discusses their therapeutic potential, and also describes emerging strategies for targeting these viruses. PMID- 17139287 TI - Targeting the Hedgehog pathway in cancer. AB - Several key signalling pathways, such as Hedgehog, Notch, Wnt and BMP-TGFbeta Activin (bone morphogenetic protein-transforming growth factor-beta-Activin), are involved in most processes essential to the proper development of an embryo. It is also becoming increasingly clear that these pathways can have a crucial role in tumorigenesis when reactivated in adult tissues through sporadic mutations or other mechanisms. We will focus here on the Hedgehog pathway, which is abnormally activated in most basal cell carcinomas, and discuss potential therapeutic opportunities offered by the progress made in understanding this signalling pathway. PMID- 17139289 TI - Mentors, influences and role models. PMID- 17139288 TI - Innovative approaches to anti-arrhythmic drug therapy. AB - Normal cardiac function requires an appropriate and regular beating rate (cardiac rhythm). When the heart rhythm is too fast or too slow, cardiac function can be impaired, with derangements that vary from mild symptoms to life-threatening complications. Irregularities, particularly those involving excessively fast or slow rates, constitute cardiac 'arrhythmias'. In the past, drug treatment of cardiac arrhythmias has proven difficult, both because of inadequate effectiveness and a risk of serious complications. However, a variety of recent advances have opened up exciting possibilities for the development of novel and superior approaches to arrhythmia therapy. This article will review recent progress and future prospects for treating two particularly important cardiac arrhythmias: atrial fibrillation and ventricular fibrillation. PMID- 17139290 TI - The future of neo-eugenics. Now that many people approve the elimination of certain genetically defective fetuses, is society closer to screening all fetuses for all known mutations? PMID- 17139291 TI - Delivering on the promise of human stem-cell research. What are the real barriers? PMID- 17139292 TI - Bad science in the headlines. Who takes responsibility when science is distorted in the mass media? PMID- 17139293 TI - Shedding a negative image. Research into their mechanism of infectivity reveals that prions might have important biological roles. PMID- 17139297 TI - Genome-wide natural antisense transcription: coupling its regulation to its different regulatory mechanisms. AB - Many genomic loci contain transcription units on both strands, therefore two oppositely oriented transcripts can overlap. Often, one strand codes for a protein, whereas the transcript from the other strand is non-encoding. Such natural antisense transcripts (NATs) can negatively regulate the conjugated sense transcript. NATs are highly prevalent in a wide range of species--for example, around 15% of human protein-encoding genes have an associated NAT. The regulatory mechanisms by which NATs act are diverse, as are the means to control their expression. Here, we review the current understanding of NAT function and its mechanistic basis, which has been gathered from both individual gene cases and genome-wide studies. In parallel, we survey findings about the regulation of NAT transcription. Finally, we hypothesize that the regulation of antisense transcription might be tailored to its mode of action. According to this model, the observed relationship between the expression patterns of NATs and their targets might indicate the regulatory mechanism that is in action. PMID- 17139300 TI - The business of science. PMID- 17139298 TI - Epigenetic mechanisms governing seed development in plants. AB - Seed development in flowering plants is initiated by the fusion of two male gametes with two female gametes--the egg cell and the central cell--which leads to the formation of an embryo and an endosperm, respectively. Fertilization independent seed formation is actively repressed by the FERTILIZATION-INDEPENDENT SEED (FIS) Polycomb group (PcG) proteins, an evolutionarily conserved class of proteins that ensures the stable transmission of developmental decisions. The FIS proteins act together in a complex and modify their target genes by applying repressive methylation on histone H3 lysine 27. In addition to its function before fertilization, the FIS complex restricts endosperm proliferation. This function is likely to be achieved by imprinting the maternal alleles of FIS target genes. However, imprinting in the endosperm is controlled not only by the FIS complex but also by DNA methylation, and the interconnections between these two processes are now being investigated. PMID- 17139302 TI - From lab to boardroom. PMID- 17139301 TI - University spin-offs: opportunity or challenge? PMID- 17139304 TI - Organic transistors: a polarized response. PMID- 17139305 TI - Geometric simulations: a lesson from virtual zeolites. PMID- 17139306 TI - Transition metal oxides: ferroelectricity driven by orbital order. PMID- 17139307 TI - High-pressure silica: densification in two steps. PMID- 17139308 TI - Material witness: pulling a leg. PMID- 17139309 TI - Tuning into the genetic orchestra using microarrays: limitations of DNA microarrays in clinical practice. PMID- 17139311 TI - A step in the right direction. PMID- 17139312 TI - We need a geriatric assessment for oncologists. PMID- 17139313 TI - Do we have a clinically useful breast cancer risk reduction agent for healthy women? PMID- 17139314 TI - Do older women with breast cancer benefit from adjuvant chemotherapy? PMID- 17139315 TI - How does use of endoscopic ultrasonography affect outcome of esophageal cancer? PMID- 17139316 TI - Does a new model improve decisions about mismatch-repair genetic testing and Lynch syndrome identification? PMID- 17139317 TI - Post-therapy changes in PSA as an outcome measure in prostate cancer clinical trials. AB - To the investigator and clinician, prostate-specific antigen (PSA) level is a seemingly perfect outcome measure because it is easily assessable, quantitative, reproducible, and inexpensive. Whether post-therapy decline in PSA reflects true clinical benefit, and whether post-therapy declines can be used as an intermediate endpoint for accelerated drug approval is still open to question. At present, no drug has been approved strictly on the basis of a post-treatment decline in PSA, as it is unproven that such PSA changes are surrogates for true clinical benefits. Post-therapy PSA changes have been associated with improved survival in patients with castrate metastatic disease. The role of PSA changes as potential surrogates of clinical benefit have only been explored to a limited degree because to date, only two prospective randomized trials showing a survival benefit have been reported. Such trials are necessary, but not a sufficient pre requisite to explore the potential role of any outcome measure as an intermediate endpoint. The clear demonstration that a post-therapy PSA change can account for all of the treatment effects seen is not yet available. A cytotoxic drug that does not produce any PSA decline is unlikely to be effective, but the converse is not always true because not all PSA rises represent a treatment failure. It is important to recognize that there are a range of clinical benefits to patients that can improve the quality and possibly the duration of survival, independent of PSA. PMID- 17139319 TI - Drug insight: vascular disrupting agents and angiogenesis--novel approaches for drug delivery. AB - Vascular disrupting agents (VDAs), or endothelial disrupting agents, attempt to exploit the vascular endothelium that supplies rapidly dividing neoplasms. Unlike antiangiogenesis agents (e.g. the monoclonal antibody bevacizumab; and tyrosine kinase inhibitors sorafenib and sunitinib) that disrupt endothelial cell survival mechanisms and the development of a new tumor blood supply, VDAs are designed to disrupt the already established abnormal vasculature that supports tumors, by targeting their dysmorphic endothelial cells. Tumor vascular endothelium is characterized by its increased permeability, abnormal morphology, disorganized vascular networks, and variable density. VDAs induce rapid shutdown of tumor blood supply, causing subsequent tumor death from hypoxia and nutrient deprivation. The safety profile of this class of compounds is more indicative of agents that are indeed 'vascularly' active. For example, VDAs can cause: acute coronary and other thrombophlebitic syndromes; alterations in blood pressure, heart rate, and ventricular conduction; transient flush and hot flashes; neuropathy; and tumor pain. Despite these cardiovascular concerns some patients have benefited from VDAs in early clinical trials. Further drug development of VDAs must include the combination of these agents with other novel biological agents, cytotoxic chemotherapy, and radiotherapy. Close monitoring of patients receiving VDAs for any cardiovascular toxicity is imperative. PMID- 17139318 TI - Increased intensity lymphodepletion and adoptive immunotherapy--how far can we go? AB - In a recent clinical trial involving patients with metastatic melanoma, immunosuppressive conditioning with fludarabine and cyclophosphamide resulted in a 50% response rate in robust long-term persistence of adoptively transferred T cells. Experimental findings indicate that lymphodepletion prior to adoptive transfer of tumor-specific T lymphocytes plays a key role in enhancing treatment efficacy by eliminating regulatory T cells and competing elements of the immune system ('cytokine sinks'). Newly emerging animal data suggest that more profound lymphoablative conditioning with autologous hematopoetic stem-cell rescue might further enhance treatment results. Here we review recent advances in adoptive immunotherapy of solid tumors and discuss the rationale for lymphodepleting conditioning. We also address safety issues associated with translating experimental animal results of total lymphoid ablation into clinical practice. PMID- 17139320 TI - Platelet-derived growth factor receptor-beta in Gorham's disease. AB - BACKGROUND: A 17-year-old male presented with pain in his lower-left chest. He had no significant medical history and was previously in good health. He had a fractured ninth left anterior rib and the tenth, eleventh and twelfth ribs were absent, which was thought to be a congenital anomaly. Several months later, he presented again with back pain, an enlarging mass in the lower-left chest wall, erosion of the lateral pedicles of the lower thoracic vertebrae and pleural effusion. INVESTIGATIONS: Physical examination, chest X-ray, MRI of the spine, incisional biopsy, serial CT imaging of the hemithorax, immunohistochemistry, flow cytometry, and enzyme-linked immunosorbent assays. DIAGNOSIS: Gorham's lymphangiomatosis with expression of platelet-derived growth factor receptor-beta and elevated circulating platelet-derived growth factor-BB. MANAGEMENT: Spine stabilization, thalidomide, celecoxib, interferon-alpha2b, pamidronate, zoledronate, thoracotomy, pleurectomy, talc pleurodesis, and imatinib mesylate. PMID- 17139321 TI - Minimal hepatic toxicity of Onyx-015: spatial restriction of coxsackie-adenoviral receptor in normal liver. AB - We administered an adenoviral vector, Onyx-015, into the hepatic artery of patients with metastatic colorectal cancer involving the liver. Thirty-five patients enrolled in this multi-institutional phase I/II trial received up to eight arterial infusions of up to 2 x 10(12) viral particles. Hepatic toxicity was the primary dose-limiting toxicity observed in preclinical models. However, nearly 200 infusions of this adenoviral vector were administered directly into the hepatic artery without significant toxicity. Therefore, we undertook this analysis to determine the impact of repeated adenoviral exposure on hepatic function. Seventeen patients were treated at our institution, providing a detailed data set on the changes in hepatic function following repeated exposure to adenovirus. No changes in hepatic function occurred with the first treatment of Onyx-015 among these patients. Transient increases in transaminase levels occurred in one patient starting with the second infusion and transient increases in bilirubin was observed in two patients starting with the fifth treatment. These changes occurred too early to be explained by viral-mediated lysis of hepatocytes. In addition, viremia was observed starting 3-5 days after the viral infusion in half of the patient, but was not associated with hepatic toxicity. To further understand the basis for the minimal hepatic toxicity of adenoviral vectors, we evaluated the replication of adenovirus in primary hepatocytes and tumor cells in culture and the expression of the coxsackie-adenoviral receptor (CAR) in normal liver and colon cancer metastatic to the liver. We found that adenovirus replicates poorly in primary hepatocytes but replicates efficiently in tumors including tumors derived from hepatocytes. In addition, we found that CAR is localized at junctions between hepatocytes and is inaccessible to hepatic blood flow. CAR is not expressed on tumor vasculature but is expressed on tumor cells. Spatial restriction of CAR to the intercellular space in normal liver and diminished replication of adenovirus in hepatocytes may explain the minimal toxicity observed following repeated hepatic artery infusions with Onyx-015. PMID- 17139322 TI - How flies get their size: genetics meets physiology. AB - Body size affects important fitness variables such as mate selection, predation and tolerance to heat, cold and starvation. It is therefore subject to intense evolutionary selection. Recent genetic and physiological studies in insects are providing predictions as to which gene systems are likely to be targeted in selecting for changes in body size. These studies highlight genes and pathways that also control size in mammals: insects use insulin-like growth factor (IGF) and Target of rapamycin (TOR) kinase signalling to coordinate nutrition with cell growth, and steroid and neuropeptide hormones to terminate feeding after a genetically encoded target weight is achieved. However, we still understand little about how size is actually sensed, or how organ-intrinsic size controls interface with whole-body physiology. PMID- 17139323 TI - Genetics of autoimmune diseases--disorders of immune homeostasis. AB - In the past few years, our extensive knowledge of the mammalian immune system and our increasing ability to understand the genetic causes of complex human disease have opened a window onto the pathways that lead to autoimmune disorders. In addition to the well-established role of genetic variation that affects the major histocompatibility complex, a number of rare and common variants that affect a range of immunological pathways are now known to have important influences on the phenotypic diversity that is seen among autoimmune diseases. Recent studies have also highlighted a previously unanticipated interplay between the innate and adaptive immune system, providing a new direction for research in this field. PMID- 17139324 TI - Signature-tagged mutagenesis: barcoding mutants for genome-wide screens. AB - DNA signature tags (molecular barcodes) facilitate functional screens by identifying mutants in mixed populations that have a reduced or increased adaptation to a particular environment. Many innovative adaptations and refinements in the technology have been described since its original use with Salmonella; they have yielded a wealth of information on a broad range of biological processes--mainly in bacteria, but also in yeast and other fungi, viruses, parasites and, most recently, in mammalian cells. By combining whole genome microarrays and comprehensive ordered libraries of mutants, high throughput functional screens can now be achieved on a genomic scale. PMID- 17139325 TI - Human laminopathies: nuclei gone genetically awry. AB - Few genes have generated as much recent interest as LMNA, LMNB1 and LMNB2, which encode the components of the nuclear lamina. Over 180 mutations in these genes are associated with at least 13 known diseases--the laminopathies. In particular, the study of LMNA, its products and the phenotypes that result from its mutation have provided important insights into subjects ranging from transcriptional regulation, the cell biology of the nuclear lamina and mechanisms of ageing. Recent studies have begun the difficult task of correlating the genotypes of laminopathies with their phenotypes, and potential therapeutic strategies using existing drugs, modified oligonucleotides and RNAi are showing real promise for the treatment of these diseases. PMID- 17139326 TI - Heredity before genetics: a history. AB - Two hundred years ago, biologists did not recognize that there was such a thing as 'heredity'. By the 1830s, however, insights from medicine and agriculture had indicated that something is passed from generation to generation, creating the context for the brilliant advances of Mendel and Darwin. Recent work on the history and philosophy of science has shed light on how seventeenth-, eighteenth- and nineteenth-century thinkers sought to understand similarities between parents and offspring. PMID- 17139327 TI - The balance between heritable and environmental aetiology of human disease. AB - The Human Genome Project and the ensuing International HapMap Project were largely motivated by human health issues. But the distance from a DNA sequence variation to a novel disease gene is considerable; for complex diseases, closing this gap hinges on the premise that they arise mainly from heritable causes. Using cancer as an example of complex disease, we examine the scientific evidence for the hypothesis that human diseases result from interactions between genetic variants and the environment. PMID- 17139328 TI - Defining the spectrum of genome policy. AB - Many achievements in the genome sciences have been facilitated by policies that have prioritized genome research, secured funding and raised public and health professional awareness. Such policies should address ethical, legal and social concerns, and are as important to the scientific and commercial development of the field as the science itself. On occasion, policy issues take precedence over science, particularly when impasses are encountered or when public health or money is at stake. Here we discuss the spectrum of current issues and debates in genome policy, and how to actively engage all affected stakeholders to promote effective policy making. PMID- 17139329 TI - Adipocyte differentiation from the inside out. AB - Improved knowledge of all aspects of adipose biology will be required to counter the burgeoning epidemic of obesity. Interest in adipogenesis has increased markedly over the past few years with emphasis on the intersection between extracellular signals and the transcriptional cascade that regulates adipocyte differentiation. Many different events contribute to the commitment of a mesenchymal stem cell to the adipocyte lineage including the coordination of a complex network of transcription factors, cofactors and signalling intermediates from numerous pathways. PMID- 17139330 TI - Powering membrane traffic in endocytosis and recycling. AB - Early in evolution, the diversification of membrane-bound compartments that characterize eukaryotic cells was accompanied by the elaboration of molecular machineries that mediate intercompartmental communication and deliver materials to specific destinations. Molecular motors that move on tracks of actin filaments or microtubules mediate the movement of organelles and transport between compartments. The subjects of this review are the motors that power the transport steps along the endocytic and recycling pathways, their modes of attachment to cargo and their regulation. PMID- 17139331 TI - Membrane-protein topology. AB - In the world of membrane proteins, topology defines an important halfway house between the amino-acid sequence and the fully folded three-dimensional structure. Although the concept of membrane-protein topology dates back at least 30 years, recent advances in the field of translocon-mediated membrane-protein assembly, proteome-wide studies of membrane-protein topology and an exponentially growing number of high-resolution membrane-protein structures have given us a deeper understanding of how topology is determined and of how it evolves. PMID- 17139333 TI - Replisome assembly and the direct restart of stalled replication forks. AB - Failure to reactivate either stalled or collapsed replication forks is a source of genomic instability in both prokaryotes and eukaryotes. In prokaryotes, dedicated fork repair systems that involve both recombination and replication proteins have been identified genetically and characterized biochemically. Replication conflicts are solved through several pathways, some of which require recombination and some of which operate directly at the stalled fork. Some recent biochemical observations support models of direct fork repair in which the removal of the blocking template lesion is not always required for replication restart. PMID- 17139332 TI - Editor meets silencer: crosstalk between RNA editing and RNA interference. AB - The most prevalent type of RNA editing is mediated by ADAR (adenosine deaminase acting on RNA) enzymes, which convert adenosines to inosines (a process known as A-->I RNA editing) in double-stranded (ds)RNA substrates. A-->I RNA editing was long thought to affect only selected transcripts by altering the proteins they encode. However, genome-wide screening has revealed numerous editing sites within inverted Alu repeats in introns and untranslated regions. Also, recent evidence indicates that A-->I RNA editing crosstalks with RNA-interference pathways, which, like A-->I RNA editing, involve dsRNAs. A-->I RNA editing therefore seems to have additional functions, including the regulation of retrotransposons and gene silencing, which adds a new urgency to the challenges of fully understanding ADAR functions. PMID- 17139334 TI - Combinatorial microscopy. AB - By taking advantage of combinations of the many rich properties of photons, new forms of optical microscopy can now be used to visualize features of samples beyond thickness and density variations. We are now within reach of viewing the motions, orientations, binding kinetics and specific transient associations of previously 'submicroscopic' cellular structures and single molecules. PMID- 17139335 TI - Functional and quantitative proteomics using SILAC. AB - Researchers in many biological areas now routinely characterize proteins by mass spectrometry. Among the many formats for quantitative proteomics, stable-isotope labelling by amino acids in cell culture (SILAC) has emerged as a simple and powerful one. SILAC removes false positives in protein-interaction studies, reveals large-scale kinetics of proteomes and - as a quantitative phosphoproteomics technology - directly uncovers important points in the signalling pathways that control cellular decisions. PMID- 17139336 TI - Characteristics of Marshallese with Type 2 Diabetes on Oahu: A Pilot Study to Implement a Community-Based Diabetic Health Improvement Project. AB - OBJECTIVES: To determine the feasibility of a resident physician-based, culturally appropriate method of decreasing the disease burden of Type 2 Diabetes Mellitus (DM2) in a group of Pacific Islanders, Marshallese living in Hawai'i. METHODS: Thirty one Marshallese with diabetes who live on the island of Oahu, Hawaii were recruited. Baseline health status of the participants was characterized. Health parameters included HgbA1c, random blood sugar (RBS), lipid panels, body mass index (BMI), blood pressure, and medical history, along with qualitative information. A focus group was held with participants prior to beginning the curriculum to determine cultural views on diabetes, health, treatment, and to identify potential obstacles to health improvement. A DM2 educational curriculum culturally relevant to Marshallese populations was then started, including instruction in lifestyle modification, adherence to medication regimens, and planned quarterly assessment of health improvement. RESULTS: Baseline quantitative analysis revealed Marshallese with diabetes to be obese and hyperglycemic, with average BMI of 30 kg/m(2), RBS of 285, and HgbA1c of 9.3. Qualitative analysis revealed that nearly half the participants admitted to symptoms of severe hyperglycemia. The initial focus group had a substantial turnout. Attendance rapidly declined, becoming so low that classes were eventually terminated. However, in two participants who attended more than three classes there was evidence of major improvements in HgbA1c, cholesterol, and qualitative markers, which were sustained after one year. CONCLUSIONS: This pilot study of Marshallese with diabetes on Oahu showed that the majority had poor glycemic control with secondary co-morbid conditions. Although many barriers exist for successful implementation of a diabetes health improvement project in this group, the groundwork for translation of this project to the Republic of Marshall Islands (RMI) has been laid; curriculum translation and patient recruitment is currently underway to advance the project. PMID- 17139338 TI - The Istanbul International Congress and the ongoing building of our field. PMID- 17139337 TI - Development of progressive oral presentations in a therapeutics course series. AB - OBJECTIVES: To develop and implement a series of progressive evidence-based, oral presentation activities within the therapeutics series to strengthen interprofessional oral communication and literature evaluation skills. DESIGN: A step-wise progression of oral presentations was created to establish interprofessional communication skills and reinforce such skills in successive modules. Students progressed from a basic oral presentation to a brief clinical trial presentation, followed by a full journal club, culminating with a therapeutic debate. Guidelines and assessment tools were developed for each presentation focusing on style, content and organization, and analytic approach. Feedback was obtained from students and faculty members through the administration of survey instruments at the midpoint of advanced pharmacy practice experiences (APPE). ASSESSMENT: The majority of students successfully completed each oral presentation. Most felt prepared for APPE presentations due to increased Microsoft PowerPoint skills, literature exposure/evaluation, and comfort/confidence in presenting before an audience. Faculty members stated that the innovation organized students' thoughts and increased presentation clarity, resulting in less need to offer student assistance during APPEs. CONCLUSION: Our progressive approach to oral presentations has been successful and well received by students and faculty members. Our innovation has fostered oral interprofessional communication skills in our pharmacy students, and we will continue to utilize this approach. PMID- 17139339 TI - Early detection of schizophrenia: current evidence and future perspectives. AB - RESEARCH INTO THE EARLY COURSE OF SCHIZOPHRENIA HAS IDENTIFIED A PREPSYCHOTIC PRODROMAL STAGE (MEAN DURATION: 4.8 years) and a psychotic prephase (mean duration: 1.3 years). Comparisons of individually matched samples have demonstrated prodromal symptoms common to schizophrenia and moderate to severe depression. It is not until positive symptoms emerge that psychosis and mood disorders become distinguishable from each other. In both disorders the prodromal stage early produces functional impairment and related social consequences. Hence, early intervention is of great public health relevance. This intervention is targeted at manifest symptoms and not at the underlying, still unknown disease process. Cognitive-behavioural therapy at the prepsychotic prodromal stage seems to favourably influence the short-term illness course. In the psychotic prephase, a combination with low-dose antipsychotics seems to have some efficacy. The aim of early recognition by the instruments discussed in this paper is to permit the identification of the largest possible proportion of at-risk persons as early as possible and their referral to appropriate treatment. PMID- 17139340 TI - Quality of life in mental disorders: challenges for research and clinical practice. AB - The term quality of life (Qol) has become a rallying cry for all those who strive to integrate patients' subjective experience of their life during illness into clinical care. With its intuitive appeal, Qol seems to be understood by everyone involved in managing health and disease. However, when examining the ever increasing research literature, it becomes clear that many methodological questions still beset this field, since neither a commonly accepted definition nor a gold standard for measuring Qol exist. On the contrary, one is irritated by the myriad of different instruments and it remains questionable how valid and comparable research results obtained with different instruments are. It is argued that, especially when attempting to measure Qol in mental disorders: a) in addition to "subjective", also "objective" (i.e., external) assessment is needed; b) the inclusion of psychopathological symptoms in Qol instruments has to be controlled for; c) in addition to well-being and satisfaction, also functioning and environmental assets have to be assessed; d) different life areas have to be considered separately, and e) changes over time have to be taken into account. A concluding section in the paper reflects on the still largely missing transfer of research results into clinical care. PMID- 17139341 TI - Should general psychiatry ignore somatization and hypochondriasis? AB - This paper examines the tendency for general psychiatry to ignore somatization and hypochondriasis. These disorders are rarely included in national surveys of mental health and are not usually regarded as a concern of general psychiatrists; yet primary care doctors and other physicians often feel let down by psychiatry's failure to offer help in this area of medical practice. Many psychiatrists are unaware of the suffering, impaired function and high costs that can result from these disorders, because these occur mainly within primary care and secondary medical services. Difficulties in diagnosis and a tendency to regard them as purely secondary phenomena of depression, anxiety and related disorders mean that general psychiatry may continue to ignore somatization and hypochondriasis. If general psychiatry embraced these disorders more fully, however, it might lead to better prevention and treatment of depression as well as helping to prevent the severe disability that may arise in association with these disorders. PMID- 17139342 TI - Psychiatric rehabilitation today: an overview. AB - All patients suffering from severe and persistent mental illness require rehabilitation. The goal of psychiatric rehabilitation is to help disabled individuals to develop the emotional, social and intellectual skills needed to live, learn and work in the community with the least amount of professional support. The overall philosophy of psychiatric rehabilitation comprises two intervention strategies. The first strategy is individual-centered and aims at developing the patient's skills in interacting with a stressful environment. The second strategy is ecological and directed towards developing environmental resources to reduce potential stressors. Most disabled persons need a combination of both approaches. The refinement of psychiatric rehabilitation has achieved a point where it should be made readily available for every disabled person. PMID- 17139343 TI - Caveats for psychiatric rehabilitation. PMID- 17139344 TI - The underutilization of psychiatric rehabilitation. PMID- 17139345 TI - The diffusion of two successful rehabilitation models. PMID- 17139346 TI - Identifying psychiatric rehabilitation interventions: an evidence and value based practice. PMID- 17139347 TI - Psychosocial rehabilitation and severe mental disorders: a public health approach. PMID- 17139348 TI - Psychiatric rehabilitation in the era of globalization. PMID- 17139349 TI - Psychiatric rehabilitation and its present role in developing countries. PMID- 17139350 TI - A plea for the 22 UN standard rules and a request to redefine psychiatry. PMID- 17139351 TI - Psychiatric rehabilitation today:an African perspective. PMID- 17139352 TI - Risk factors for schizophrenia. Follow-up data from the Northern Finland 1966 Birth Cohort Study. AB - This paper updates single risk factors identified by the Northern Finland 1966 Birth Cohort Study up to the end of year 2001 or age 34. Impaired performance (e.g., delayed motor or intellectual development) or adverse exposures (e.g., pregnancy and birth complications, central nervous system diseases) are associated with an increased risk for schizophrenia. However, upper social class girls and clever schoolboys also have an increased risk to develop schizophrenia, contrasted to their peers. Individuals who subsequently develop schizophrenia follow a developmental trajectory that partly and subtly differs from that of the general population; this trajectory lacks flexibility and responsiveness compared to control subjects, at least in the early stages. We propose a descriptive, lifespan, multilevel systems model on the development and course of schizophrenia. PMID- 17139353 TI - The relationships between depression and remission in first-episode psychosis. AB - This study assessed changes in depressive symptoms over time in 57 patients with first-episode psychosis, and investigated the relationships of these symptoms during the acute psychotic episode and the post-psychotic period with treatment outcome. Assessment instruments included the Calgary Depression Scale (CDS) and the Positive and Negative Syndrome Scale (PANSS). For the evaluation of treatment outcome, recently proposed operational remission criteria were used. PANSS factor analysis identified a depression/anxiety factor (PANSSD/ A) at baseline, which separated into "pure" depression (PANSS-D) and anxiety (PANSS-A) factors at 24 months. There were strong correlations between the CDS and the PANSS-D/A, PANSS-D and PANSS-A scores at baseline, but at 24 months significance was lost between CDS and PANSS-A. Compared to non-remitters, patients who achieved remission had significantly higher baseline CDS scores, but depressive symptoms resolved with antipsychotic treatment. Non-remitting patients had relatively low baseline CDS scores, but their depressive symptoms persisted throughout the study. These findings suggest that depressive symptoms in the acute psychotic episode differ from those in the post-psychotic period in terms of their phenomenology, temporal relationship to psychosis, and treatment response. PMID- 17139354 TI - Understanding of the term "schizophrenia"by the British public. AB - A postal survey of a representative sample of UK adults was conducted. Subjects were asked "What do you understand by the term 'schizophrenia'?".Four hundred four completed questionnaires were received (81% response rate). Forty-two percent of respondents mentionedat least one first rank symptom of schizophrenia or gave a description that reasonably approximated to any diagnostic feature as stated inICD-10. Forty percent mentioned "split" or "multiple" personality. Thirty-eight percent described auditory hallucinations or "hearing voices".Fifteen percent mentioned "delusions" or described passivity experiences. Only 6% of subjects mentioned violence in their descriptions. PMID- 17139355 TI - World Health Organization's Mental Health Atlas 2005:implications for policy development. AB - In 2005, the World Health Organization (WHO) launched the second edition of the Mental Health Atlas, consisting of revised and updated information on mental health from countries. The sources of information included the mental health focal points in the Ministries of Health, published literature and unpublished reports available to WHO. The results show that global mental health resources remain low and grossly inadequate to respond to the high level of need. In addition, the revised Atlas shows that the improvements over the period 2001 to 2004 are very small. Imbalances across income groups of countries remain largely the same. Enhancement in resources devoted to mental health is urgently needed, especially in low- and middle-income countries. PMID- 17139356 TI - Ethics in psychiatry: a framework. AB - Defining bioethics as the rational use of dialogue in the formulation, justification, and application of ethical principles, with the aim ofgenerating good practices in research, clinical practice, and advocacy, this paper focuses on methods for bioethical deliberation relevantto psychiatry. Stressing that bioethics fuses the two main ethical traditions in Western thought, the deontological and the teleological, thepaper emphasizes the three conditions that any intervention, if considered in the context of bioethics, should fulfil: it should be appropriateto the problem at hand, it should be good (in the sense that it does good to those who receive it but also to those who perform it),and it should be just (in the sense that its outcomes can be generalized to the whole of society). Some implications of these notions for thepractice and teaching of psychiatry are presented. PMID- 17139357 TI - Letter to the editor. PMID- 17139358 TI - An agenda for change: the role of the WPA in global psychiatric education. PMID- 17139359 TI - Advancement of scientific knowledge through international collaborative research. PMID- 17139360 TI - Plasma peptidome: A new approach for assessing thrombotic risk? PMID- 17139361 TI - PAI-1 inhibition in obesity and the metabolic syndrome: a promising therapeutic strategy. PMID- 17139362 TI - Prevention of distal embolization in patients undergoing mechanical revascularization for acute myocardial infarction. A review of current status. AB - Distal embolization is a relatively common complication in primary angioplasty and is associated with poor perfusion and higher mortality. The aim of this article is to critically review literature on pharmacological and mechanical therapies to prevent distal embolization in patients undergoing primary angioplasty. The literature was scanned by formal searches of electronic databases (MEDLINE, Pubmed) from January 1990 to March 2006 and scientific session abstracts (from January 1990 to March 2006) and oral presentation and/or expert slide presentations (from January 2002 to March 2006) (on TCT, AHA, ESC, ACC and EuroPCR websites). No language restrictions were enforced. Several pharmacological and mechanical therapies have been investigated to prevent distal embolization. Abciximab has been shown to reduce mortality, and its early administration may provide additional benefits in outcome due to improvement in preprocedural reperfusion. The results of randomized trials on adjunctive mechanical devices remain controversial. Even though they reduce distal embolization and improve myocardial perfusion, no benefits have been observed in terms of 30-day survival. Adjunctive abciximab has improved survival, and its early administration is to be recommended, particularly when transportation to a primary PCI center is needed. Pending the results of large randomized trials with long-term follow-up data, the routine use of adjunctive mechanical devices to prevent distal embolization cannot be recommended, though selective use of these devices might be considered when large thrombotic burden is present. PMID- 17139363 TI - Combined partial exon skipping and cryptic splice site activation as a new molecular mechanism for recessive type 1 von Willebrand disease. AB - We describe the complex picture associated with a mutated splice junction in intron 13 of von Willebrand factor (VWF) gene. The proband, characterized by a marked decrease in plasma and platelet VWF and near normal multimer organization, was classified as recessive type 1 von Willebrand disease (VWD). Genetic analysis demonstrated that he was homozygous for the 1534-3C > A mutation in the consensus sequence of the acceptor splicing site of intron 13 of the VWF gene. Platelet mRNA analysis documented three VWF transcripts: a wild type generated by the correct recognition of the mutated splice site, a smaller transcript not containing exon 14, and a longer one that, in addition to exons 13 and 14, included a 62bp fragment corresponding to the end of intron 13. The small transcript derives from the skipping of exon 14, the long one from the activation of a cryptic splice site in intron 13; both show a premature stop codon in VWF propeptide, so the proband VWF derives entirely from the correct splice site recognition. Combined incomplete exon skipping and cryptic splice site activation are first recognized in VWD. Since the 1534-3C > A mutation does not abolish the normal processing of mRNA, it is unlikely to be found in type 3 VWD. This mutation therefore appears to be peculiar to type 1 VWD. PMID- 17139364 TI - Differential effects of the loss of intrachain- versus interchain-disulfide bonds in the cystine-knot domain of von Willebrand factor on the clinical phenotype of von Willebrand disease. AB - Von Willebrand factor (VWF) contains a large number of cysteine residues, which all form disulfide bonds. Mutations of cysteines located in the cystine-knot (CK) domain of VWF have been identified in both qualitative type 2A (IID) and quantitative type 3 von Willebrand disease (VWD). Our objective was to test the hypothesis that the difference in phenotype is related to whether the mutated cysteine residue is involved in either interchain- or intrachain-disulfide-bond formation. The effects of three cysteine mutations which are all located in the CK-domain of VWF, C2773S (type 2A(IID)), C2739Y (type 3), and C2754W (type 3), were studied by transient expression in 293T cells. Cotransfection of wild-type (wt) and C2773S VWF constructs reproduced the plasma phenotype of heterozygous type 2A(IID) patients, with normal to high levels of VWF antigen (VWF:Ag), absence of high-molecular-weight multimers, and the presence of intervening bands between the normal multimers. In contrast, single transfections of C2739Y or C2754W resulted in a quantitative VWF defect with low VWF:Ag levels, and co transfections of wt and mutant constructs resulted in a 50% reduction of VWF:Ag and only a minor effect on VWF multimerization. We demonstrated N-terminal dimerization of VWF-C2773S and both N- and C-terminal dimerization of VWF-C2754W. Our data suggest that loss of a single disulfide bond in the CK-domain of VWF leads to a recessive, quantitative VWF deficiency if an intrachain-disulfide bond is involved, and to a dominant-negative, qualitative defect of VWF if an interchain-disulfide bond is involved. PMID- 17139365 TI - SELDI-TOF plasma profiles distinguish individuals in a protein C-deficient family with thrombotic episodes occurring before age 40. AB - We tested the hypothesis that differences in the low-molecular-weight (500-20,000 Da) proteomic profile of plasma may be detectable between members of a protein C deficient family who have suffered thrombotic events before age 40 compared to family members without a history of venous thrombosis. Unfractionated plasma samples from members of a previously described large thrombophilic kindred with type I protein C deficiency were applied to ProteinChip weak cation exchange interaction arrays (WCX2; Ciphergen Biosystems, Fremont, CA, USA) and subjected to SELDI-TOF (surface-enhanced laser desorption/ionization time-of-flight) mass spectrometry using the Ciphergen PBSII ProteinChip System (Ciphergen Biosystems). Profiles were analyzed by a boosted decision-tree algorithm. When individuals who had presented with deep venous thrombosis (DVT) before the age of 40 (n = 21) were compared to age-matched, healthy family members (n = 50), the proteomic patterns defined by the decision-tree analysis could classify the entity of DVT before age 40 with 67% sensitivity, at a specificity of 86%. When a small group of cases with history of superficial venous thrombosis (n = 6) was added to the case group, the sensitivity was 87.5% at a specificity of 80%. These data support the hypothesis that members of the protein C deficient Vermont kindred II who suffer a thrombotic event before age 40 display significant differences in low molecular-weight proteomics profile compared to those who remain disease-free. This is the first study to apply SELDI-TOF technology in conjunction with a bioinformatics tool to analyze low-molecular-weight proteomic patterns in patients with venous thrombosis. PMID- 17139366 TI - Tiplaxtinin impairs nutritionally induced obesity in mice. AB - To investigate the effect of tiplaxtinin, designed as a synthetic inhibitor of plasminogen activator inhibitor-1 (PAI-1), on obesity, male C57Bl/6 mice (13-14 weeks old) were kept on a high-fat diet (20.1 kJ/g) for four weeks without or with addition of tiplaxtinin (PAI-039) at a dose of 2 mg/g food. At the time of sacrifice, body weights were significantly lower in the inhibitor-treated mice (p < 0.0005). The weights of the isolated subcutaneous and gonadal fat deposits were also significantly lower (both p < 0.0005), associated with adipocyte hypotrophy. Inhibitor-treated adipose tissues displayed similar blood vessel size, but a higher blood vessel density. Fasting glucose and insulin levels, as well as glucose-tolerance tests were not significantly affected by the inhibitor treatment, whereas plasma triglyceride levels were significantly reduced (p = 0.02) and LDL-cholesterol levels significantly enhanced (p = 0.0002). Insulin tolerance tests revealed significantly lower glucose levels at the end of the test in the inhibitor treated mice (p = 0.03). Thus, in this model of diet induced obesity in mice administration of tiplaxtinin resulted in impaired adipose tissue development. PMID- 17139367 TI - Effect of antiplatelet agents clopidogrel, aspirin, and cilostazol on circulating tissue factor procoagulant activity in patients with peripheral arterial disease. AB - Tissue factor (TF) is the physiological initiating mechanism for blood coagulation. Platelets play an important role in monocyte TF expression, thrombosis and inflammation. Aspirin, clopidogrel and cilostazol, which inhibit platelet responses by different mechanisms, are widely used in patients with arterial diseases. We tested the hypothesis that platelet-inhibiting agents inhibit the levels of circulating TF procoagulant activity (TF-PCA) in patients with peripheral arterial disease (PAD). Twenty-six patients with lower extremity PAD, average age 65.9 +/- 8.4 years (mean +/- SEM), were studied at baseline and following sequential two-week treatment regimens with aspirin (325 mg daily), clopidogrel (75 mg daily) or a phosphodiesterase inhibitor cilostazol (100 mg twice daily) singly, and with each possible combination of these agents. Circulating TF-PCA in whole blood, and plasma factor VIIa, prothrombin fragment F1.2, thrombin-antithrombin complexes (TAT), and P-selectin were measured. Baseline TF-PCA levels in the patients were elevated (131 +/- 19 U/ml) compared to control subjects (23 +/- 2, p < 0.0001). TF-PCA levels declined following treatment with clopidogrel alone, and with combinations of clopidogrel with aspirin or cilostazol, with the lowest levels being with the triple-drug combination. Plasma P-selectin declined in all treatment groups. No changes were noted in plasma factor VIIa, F1.2 or TAT. In conclusion, treatment of PAD patients with antiplatelet agents decreases circulating TF, a molecule with prothrombotic and proinflammatory effects. These findings suggest an unrecognized mechanism, beyond inhibiting aggregation responses, for the efficacy of antiplatelet drugs in patients with arterial diseases. PMID- 17139368 TI - Incidence of venous thromboembolism in first-degree relatives of patients with venous thromboembolism who have factor V Leiden. AB - The factor V Leiden (FVL) mutation, a genetic abnormality with an autosomal mode of inheritance, is associated with an increased risk of venous thromboembolism (VTE). We aimed to determine the annual incidence of VTE in first-degree relatives of patients with VTE and FVL and to identify factors in patients and the relatives that influence this incidence. In this retrospective and prospective cohort study, the incidence of objectively diagnosed first episodes of VTE was assessed in 553 first-degree relatives of 161 patients with acute VTE and FVL. The annual incidence of VTE was 0.43% (95% CI, 0.3 to 0.56) with FVL and 0.17% (95% CI, 0.07 to 0.27) without FVL (relative risk of 2.5,95% CI, 1.3 to 4.7). A majority (70%) of episodes of VTE were provoked, and this proportion was similar with and without FVL. A larger proportion of VTE was provoked in women (83%) that in men (33%), with the difference accounted for by pregnancy and use of oral contraceptives. The proportion of pregnancies complicated by VTE was 3.9% (95% CI, 2.0-5.8) with FVL and 1.4% (95% CI, 0.04-2.7) without FVL. FVL is associated with a two- to threefold increase in VTE in first-degree relatives of patients with VTE. No subgroup of relatives was identified who require more than routine prophylaxis because of a particularly high risk of VTE. PMID- 17139369 TI - A multi-dose pharmacokinetic study of dalteparin in haemodialysis patients. AB - Low-molecular-weight heparins undergo renal elimination, and therefore the proper dosing in hemodialysis (HD) patients is unclear. It was the objective of this study to evaluate the pharmacokinetic (PK) parameters of dalteparin in patients receiving chronic HD for end-stage renal disease. We performed a multidose PK study with prophylactic doses of dalteparin in twelve HD patients. Dalteparin 5,000 IU was administered subcutaneously daily for four consecutive days, with HD performed on day 2 and day 4. Anti-factor Xa activity was determined daily and at multiple blood samples after the 3rd and 4th dose. Eleven of 12 patients completed the study. The mean (range) PK parameters determined after the 4th dose were as follows: i) maximum concentration (Cmax ) was 0.31 IU/ml (0.06 to 0.55 IU/ml); ii) time to Cmax was 3.55 hours (2.59 to 4.96 hr); iii) area under the curve was 3.24 IU*hr/ml (0.64 to 6.44 IU*hr/ml); iv) half-life was 3.82 hr (2.03 to 9.63 hr); and v) trough anti-factor Xa activity 0.04 IU/ml (0.02 to 0.08 IU/ml). No major bleeding was observed. In general, patients with lower body weight exhibited a higher Cmax . From this pilot PK study, we have determined initial PK parameters for dalteparin in HD patients. Although a standard prophylactic dose was used, we found that in this patient population differences in body weight influenced the Cmax. Future studies to evaluate the PK parameters of dalteparin in patients receiving chronic HD may have to use weight-based dosing and will need to be performed over a longer period of time. PMID- 17139370 TI - Palmitoylation at Cys595 is essential for PECAM-1 localisation into membrane microdomains and for efficient PECAM-1-mediated cytoprotection. AB - The Ig-ITIM superfamily member, PECAM-1 acts as a negative regulator of ITAM signalling pathways in platelets involving GPVI/FcR gamma chain and Fc?RIIa. This negative feedback loop involves regulation of collagen and GPVI-dependent aggregation events, platelet-thrombus-growth on immobilised collagen under flow and Fc?RIIa-mediated platelet responses. In this study, we show that PECAM-1 is selectively palmitoylated involving a thioester linkage with an unpaired cysteine residue at amino acid position 595 in its cytoplasmic domain. As palmitoylation is known to target proteins to membrane microdomains, we investigated the microdomain localisation for PECAM-1 in platelets and nucleated cells. In unstimulated platelets, approximately 20% of PECAM-1 is localised to Triton insoluble microdomain fractions and it does not increase with platelet activation by collagen, collagen-related peptide, thrombin- or human-aggregated IgG. PECAM-1 is in close physical proximity with GPVI in platelet microdomains. Removal of platelet cytoskeleton prior to sucrose-density-gradient separation showed that PECAM-1 was associated with both the Triton-soluble and membrane skeleton in microdomain-associated fractions. Disruption of microdomains by membrane cholesterol depletion resulted in loss of PECAM-1 localisation to membrane microdomains. Mutational analysis of juxtamembrane cysteine residue to alanine (C595A) of human PECAM-1 resulted in loss of palmitoylation and a sixfold decrease in association with membrane microdomains. Functionally, the palmitoylated cysteine 595 residue is required, in part, for efficient PECAM-1 mediated cytoprotection. These results show that cysteine 595 is required for constitutive association of PECAM-1 with membrane microdomains and PECAM-1 mediated cytoprotection, where it may act as a crucial regulator of signaling and apoptosis events. PMID- 17139371 TI - Comparison of VASP-phosphorylation assay to light-transmission aggregometry in assessing inhibition of the platelet ADP P2Y12 receptor. AB - There is need to improve platelet function testing to monitor the response to antiplatelet drugs. We compared flow-cytometric analysis of intraplatelet vasodilator-stimulated phosphoprotein phosphorylation (VASP-P) to light transmission aggregometry for the detection of drug-induced in-vitro inhibition of the platelet P2Y12 ADP receptor on 22 healthy subjects (10 males, 12 females, 28.5 +/- 6.6 years). The platelet reactivity index (PRI) of VASP was calculated both from mean fluorescence intensity (MFI) and percent of fluorescence-positive platelets in the presence of PGE1 with or without ADP (10 microM). Platelet aggregation was induced by ADP (1.25, 2.5 and 5 microM). Cangrelor, a competitive inhibitor of the P2Y12 receptor, preincubated 5 minutes, induced a concentration dependent inhibition of platelet ADP-receptor function in both tests. Indeed PRI (%) based on either MFI or percent platelets gated were highly correlated with each other (r = 0.97, p %lt; 0.0001) and with aggregation induced by ADP. The IC50 of cangrelor against each of the three ADP concentrations used in aggregometry increased from 5.8 +/- 3.4 nM to 23.1 +/- 4.0 nM and to 98 +/- 25 nM, respectively. The IC50 of cangrelor based on VASP-P was within the same range (25.5 +/- 7.7 nM). No correlation was observed between IC50 values of cangrelor and ADP concentrations giving 50% effect (EC50) in the absence of the drug. However, at 10 nM cangrelor seven subjects could be identified by the VASP-P assay as " low responders " to the drug (PRI > 50%), and six of them also had an aggregation response to 5 micro M ADP > 50%. These six subjects showed the lowest ADP EC50 values in the absence of the drug, possibly reflecting high sensitivity of their platelet P2Y12 receptors to ADP. In conclusion, both the VASP-P assay and light-transmission aggregometry detect in a comparable way in-vitro pharmacological inhibition of the platelet P2Y12 ADP receptor and its individual variability. PMID- 17139372 TI - Platelet microparticle formation and thrombin generation under high shear are effectively suppressed by a monoclonal antibody against GPIba. AB - We studied the inhibition of platelet microparticle (MP) formation and thrombin generation under high shear forces. We hypothesized that an inhibitor of the GPIb a -von Willebrand factor (vWF) interaction would be more effective in suppressing MP formation and thrombin generation than GPIIb/IIIa inhibitors. Platelet-rich plasma (PRP) anticoagulated with PPACK (D-Phe-Pro-Arg chloromethyl ketone) was exposed in a cone-and-plate viscometer (shear: 5,000 s(-1) for 5 min) in the presence of antagonists to GPIb a (the monoclonal antibody [Mab] Ib-23) or to GPIIb/IIIa (abciximab, tirofiban, eptifibatide) at their IC90 determined in platelet aggregometry with ristocetin or ADP, respectively. We used double labeling (CD41-PE and annexin-V-FITC) for flow cytometric detection of MP and their aminophospholipid exposure. Thrombin generation was measured using PRP prepared from ACD anticoagulated blood. About 40% of the thrombin generation was found to be mediated by the MP fraction of the PRP. Blockade of GPIb a with Mab Ib-23 reduced MP formation and thrombin generation by 50%, and was more effective than any GPIIb/IIIa antagonist. The combination of Mab Ib-23 with one of the GPIIb/IIIa inhibitors further reduced the MP formation to ~ 30%. The antibody also partially inhibited thrombin induced platelet aggregation. Epitope mapping suggested that Mab Ib-23 binds between the amino acids 201 and 268 of GPIb a , explaining the interference with vWF and thrombin interaction. In contrast to the commonly used GPIIb/IIIa antagonists, the blockade of GPIb a with Mab Ib-23 effectively reduces the prothrombotic MP generation and thrombin formation at shear rates typically found in arterial stenoses. PMID- 17139373 TI - Multiple electrode aggregometry: a new device to measure platelet aggregation in whole blood. AB - Several methods are used to analyse platelet function in whole blood. A new device to measure whole blood platelet aggregation has been developed, called multiple electrode platelet aggregometry (MEA). Our aim was to evaluate MEA in comparison with the single platelet counting (SPC) method for the measurement of platelet aggregation and platelet inhibition by aspirin or apyrase in diluted whole blood. Platelet aggregation induced by different concentrations of ADP, collagen and TRAP-6 and platelet inhibition by apyrase or aspirin were determined in citrateor hirudin-anticoagulated blood by MEA and SPC. MEA indicated that spontaneous platelet aggregation was lower, and stimulated platelet aggregation was higher in hirudin- than citrate-anticoagulated blood. In hirudin anticoagulated, but not citrate-anticoagulated blood, spontaneous platelet aggregation measured by MEA was inhibited by apyrase. For MEA compared with SPC the dose response-curves of agonist-induced platelet aggregation in citrate- and hirudin-blood showed similar EC50 values for TRAP, and higher EC50 values for ADP (non-significant) and collagen (p < 0.05). MEA and the SPC method gave similar results concerning platelet-inhibition by apyrase and aspirin. MEA was more sensitive than SPC to the inhibitory effect of aspirin in collagen-induced aggregation. In conclusion, MEA is an easy, reproducible and sensitive method for measuring spontaneous and stimulated platelet aggregation, and evaluating antiplatelet drugs in diluted whole blood. The use of hirudin as an anticoagulant is preferable to the use of citrate. MEA is a promising technique for experimental and clinical applications. PMID- 17139374 TI - Ethyl pyruvate exerts combined anti-inflammatory and anticoagulant effects on human monocytic cells. AB - Sepsis is characterized by a concurrent activation of inflammation and coagulation. Recently, recombinant human activated protein C was shown to decrease mortality in patients with severe sepsis presumably due to a combined anti-inflammatory and anticoagulant effect. These promising findings led to a search for other products that influence both the inflammatory and the procoagulant response to severe infection. Ethyl pyruvate (EP) was recently identified as an experimental anti-inflammatory agent during endotoxemia and sepsis. The aim of the present study was to investigate whether EP influences coagulation besides its anti-inflammatory effects. For this we investigated the effects of EP on the expression and function of tissue factor (TF), the principal initiator of coagulation activation in sepsis, in human monocytic (THP-1) cell cultures. EP dose-dependently inhibited the production of tumor necrosis factor (TNF)-alpha, macrophage inflammatory protein (MIP)-1alpha and MIP-1beta by lipopolysaccharide (LPS)-stimulated THP-1 cells at mRNA and protein level, thereby confirming its anti-inflammatory properties in this in-vitro system. In addition, EP dose-dependently attenuated the increases in TF mRNA levels, TF protein-surface expression and cell-surface-associated TF activity in LPS stimulated THP-1 cells. These results demonstrate for the first time that EP is a compound with combined anti-inflammatory and anticoagulant effects. PMID- 17139375 TI - Focal arterial inflammation is augmented in mice with a deficiency of the protein C gene. AB - Increased risk of thrombosis, with propitious conditions for fibrin deposition, along with upregulation of inflammation, are important factors that enhance plaque formation in atherosclerosis. Evidence supporting the role of anticoagulant protein C (PC) as an inflammatory agent has emerged, supplementing its well-known function as an anticoagulant. Thus, we sought to examine whether a PC deficiency would lead to an enhanced response to an acute arterial hyperplasic challenge. The presentation of early arterial inflammation was studied using a copper/silicone arterial cuff model of accelerated focal neointimal remodeling in mice with a heterozygous total deficiency of PC (PC+/-). Increased inflammation, cell proliferation, cell migration, fibrin elevation, and tissue necrosis were observed in the treated arteries of PC+/- mice, as compared to arteries of equally challenged age- and gender-matched WT mice. These results indicate that PC+/- mice subjected to this challenge displayed enhanced focal arterial inflammation and thrombosis, leading to larger neointimas and subsequent localized occlusion, as compared to their WT counterparts. PMID- 17139376 TI - A non-anticoagulant synthetic pentasaccharide reduces inflammation in a murine model of kidney ischemia-reperfusion injury. AB - Fondaparinux is a synthetic pentasaccharide that selectively inhibits factor Xa (FXa) in an antithrombin-dependent fashion. This newly developed anticoagulant is used in the prevention and treatment of venous thromboembolism. Recently, we showed that fondaparinux reduces inflammation and protects the kidney from ischemia-reperfusion (I/R) injury. However, the relative contributions of the anticoagulant and anti-inflammatory activities of fondaparinux to the observed protection is unknown. To address this, we chemically modified fondaparinux to abolish its affinity for antithrombin and analyzed the effect of this non anticoagulant (NAC)-pentasaccharide on binding of U937 cells to P-selectin in vitro and on inflammation in a murine model of kidney I/R injury. NAC pentasaccharide was as effective as fondaparinux at inhibiting the binding of U937 cells to P-selectin. In addition, NAC-pentasaccharide significantly reduced IL-6 and MIP-2 expression and injury in the kidney I/R model. These findings indicate that the anti-inflammatory activity of fondaparinux can be dissociated from its anticoagulant activity and that NAC-pentasaccharide is protective in kidney I/R injury. PMID- 17139377 TI - LFA-1 and VLA-4 involved in human high proliferative potential-endothelial progenitor cells homing to ischemic tissue. AB - Cumulative evidences have revealed that endothelial progenitor cell (EPC) transplantation can promote the neovascularization in ischemic tissue, but the mechanism of EPCs homing to the site of ischemia is poorly understood. In this study, to investigate the mechanism of human umbilical cord blood-derived high proliferative potential-endothelial progenitor cells (HPP-EPCs) homing to ischemic tissue we evaluated the expression of lymphocyte function-associated antigen-1 (LFA-1, or CD11a/CD18) and very late antigen-4 (VLA-4, or CD49d/CD29) in EPCs and the changes of expression level of their ligands, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), in ischemic tissue and performed the adhesion and migration assays to analyze the interaction between the receptors and ligands. Furthermore, we studied the roles of LFA-1 and VLA-4 in EPC homing in an ischemic model of mice. The results show that LFA-1 andVLA-4 were expressed in HPPEPCs and ICAM-1 and VCAM-1 were expressed in vessel endothelium in ischemic tissues. The pre-incubation of HPP EPCs with neutralizing antibodies against CD11a or CD49d reduced adhesion and migration of HPP-EPCs in vitro and reduced recovery of hind-limb blood flow, capillary density and incorporation of HPP-EPC into ischemic tissues in vivo. Furthermore, the pre-incubation of HPP-EPCs with the combination of CD11a and CD49d antibodies led to synergistically negative effects on adhesion and transmigration of HPP-EPCs in vitro, and on the homing of HPP-EPCs to ischemic tissue and on neovascularization capacity in vivo. These results indicate that LFA-1 andVLA-4 are involved in HPP-EPC homing to ischemic tissues. PMID- 17139378 TI - Anti-metastatic effect of a non-anticoagulant low-molecular-weight heparin versus the standard low-molecular-weight heparin, enoxaparin. AB - Low-molecular-weight heparins (LMWH) exhibit potent anticoagulant efficacy via their plasmatic effects on thrombin and factor Xa. These agents are also effective in releasing endothelial tissue factor pathway inhibitor (TFPI), the natural inhibitor of tissue factor, and exhibit significant anti-metastatic effects in experimental animal models. However, the potential for bleeding complications has slowed down the more widespread adoption of LMWH therapy in cancer patients. In this study, the effect of a non-anticoagulant form of LMWH (NA-LMWH) on experimental lung metastasis and tumor cell-induced platelet aggregation in vivo was compared to the LMWH enoxaparin. Using the B16 melanoma mouse model of metastasis, subcutaneous (s.c.) injection of NA-LMWH or enoxaparin (10 mg/kg), three hours before intravenous (i.v.) injection of metastatic melanoma cells, followed by daily doses for 14 days, reduced lung tumor formation by 70% (P < 0.001). I.v. injection of tumor cells resulted in a significant (50 62%, P < 0.01) fall in platelet counts. Pre-injection (i.v.) of enoxaparin completely abolished the tumor cell-induced thrombocytopenia, whereas NA-LMWH had no effect. Four hours after a single s.c. dose, enoxaparin but not NA-LMWH prolonged the clotting time three-fold and delayed the time to clot initiation more than 10-fold as measured by a Sonoclot analyzer and by thromboelastography, respectively. Enoxaparin but not NA-LMWH demonstrated a significant anticoagulant effect in mice. Both NA-LMWH and enoxaparin caused similar TFPI release from endothelial cells in vitro. These data provide evidence to support the potential of NA-LMWH as an anti-metastatic agent without any significant impact on coagulation. PMID- 17139379 TI - Fucosylated chondroitin sulfate as a new oral antithrombotic agent. AB - Fucosylated chondroitin sulfate is a potent anticoagulant polysaccharide extracted from sea cucumber. Its anticoagulant activity is attributed to the presence of sulfated fucose branches. We have shown that intravascular injection of fucosylated chondroitin sulfate inhibits thrombus formation in a venous and an arterial shunt model in rats. Since this compound resists digestion by enzymes that cleave mammalian glycosaminoglycans, we investigated the possibility that fucosylated chondroitin sulfate might be absorbed after oral administration. In fact, after oral administration of fucosylated chondroitin sulfate to rats, we observed a dose-dependent increase in the plasma anticoagulant activity, as assessed by assays for activated partial thromboplastin time (aPTT) and thrombin time (TT) (about 3- and 5-fold, respectively) and by anti-IIa activity. Furthermore, animals receiving daily oral doses of this glycosaminoglycan showed a decrease in thrombus weight on experimental models of venous and arterial shunt thrombosis. This antithrombotic action clearly has a strong relationship with anticoagulant activity. Similar doses of heparin administered orally had no effect on the plasma anticoagulant activity or on the thrombus weight. Finally, we observed that fucosylated chondroitin sulfate given orally to rats did not modify the bleeding time. Overall, our results indicate that fucosylated chondroitin sulfate is absorbed after oral administration and could become a promising oral anticoagulant. PMID- 17139380 TI - Chronological expression of PAR isoforms in acute liver injury and its amelioration by PAR2 blockade in a rat model of sepsis. AB - The liver can be injured and its functions altered by activation of the coagulation and inflammatory processes in sepsis. The objective of the present study was to investigate the pattern of protease- activated receptors (PARs) over time in a model of acute liver injury induced by lipopolysaccharide (LPS); and whether PARs play a role in this process and exert their effects through inflammation and coagulation. Levels of tumor necrosis factor-a (TNF-a) were significantly expressed 1 h after LPS administration followed by: i) an increase in levels of tissue factor, factor VIIa, thrombin and plasminogen activator inhibitor-1; ii) unchanged or steady levels of tissue factor pathway inhibitor; and iii) subsequent deposition of fibrin in the liver tissue, that led to the elevation of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), which are associated with liver injury. The expression of all PAR isoforms (1-4) was elevated, and each isoform had a distinct cellular localization (hepatocytes, Kupffer cells, the portal triad area, and central veins) and a time-dependent pattern of expression. The immuno-reactivity of PAR2 and 4 in Kupffer cells was intense. Interestingly, PAR2 blocking peptide improved the healing of liver injuries, an effect that was associated with suppression of TNF-a elevation, and normalization of coagulation and fibrinolysis. This ultimately led to decreased fibrin formation in the injured liver. The present study reveals a distinct chronological expression and cellular localization of PARs in LPS-mediated liver injury and shows that blockade of PAR2 may play a crucial role in treating liver injury, via normalization of inflammation, coagulation and fibrinolytic pathways. PMID- 17139381 TI - A versatile strategy for preimplantation genetic diagnosis of haemophilia A based on F8-gene sequencing. AB - Preimplantation genetic diagnosis (PGD) of hemophilia A (HA) and other X-linked diseases through sex selection implies that male embryos will be systematically discarded, even though 50% are unaffected. The objective of the present work was to develop a PGD protocol for direct mutation identification that could be applied to first polar bodies (1PBs) in several HA clinical cases. Single buccal cells from controls and patients, and 1PBs were subjected to primer extension preamplification (PEP) PCR followed by amplification of F8 gene coding and intronic flanking regions, and direct sequencing. Moreover, multiplex fluorescent amplification of four short tandem repeats was adapted to a single cell preamplification in order to rule out contamination and allele drop-out, and for confirmatory indirect diagnosis. A couple at risk of HA transmission, with a familial mutation characterized as a 41-bp duplication in exon 14 of the F8 gene, was selected for the first clinical study. After optimizing the protocol, the complete F8 gene coding sequence was obtained from single cells to demonstrate the sensitivity of our methodology although in any clinical case only the relevant region, not the whole gene, must be amplified. The woman enrolled in the first clinical case has completed the first in-vitro fertilization cycle, and seven oocytes were analyzed with concordant results by both linkage analysis and direct sequencing method. Only one oocyte, among those diagnosed as mutation free, developed to embryo at day 3. It was transferred but pregnancy was not achieved. This PGD procedure enables non-affected and noncarrier embryo selection in families with any point or small-range mutation in the F8 gene, without the need for further custom-made modifications. PMID- 17139382 TI - Satellite symposium on "Haemostasis, Vascular Biology, and Infectious Agents", September 3, 2006, Maastricht, the Netherlands. PMID- 17139383 TI - Intravenous immunoglobulin-associated vena cava thrombosis. PMID- 17139384 TI - Low-grade haemostatic activation and increased fibrin turnover due to a retained central venous guide wire, found accidentally after eight years. PMID- 17139385 TI - Factor XII gene (F12) -4C/C polymorphism in combination with low protein S activity is associated with deep vein thrombosis. PMID- 17139386 TI - Classification of patients with antiphospholipid syndrome into risk categories: an evolving process. PMID- 17139387 TI - Monocyte responsiveness towards different arteriogenic stimuli: a functional comparison of various chemoattractants and their combinations. PMID- 17139388 TI - Exercise increases tissue-type plasminogen activator expression in rat cardiomyocytes. PMID- 17139389 TI - No effect of oral hormone replacement therapy on platelet function in postmenopausal women with coronary artery disease. PMID- 17139390 TI - EMG vagus nerve stimulator artifact. AB - EMG artifact produced by a VNS stimulator is described. A patient with a VNS stimulator underwent an EMG study for suspected ALS. Artifacts that appeared similar to positive sharp waves or fibrillations were noted that could produce a false clinical diagnosis. These VNS-EMG artifacts matched well with the VNS generator's set parameters. We conclude that EMG findings must be interpreted with caution in patients with VNS implants and also that EMG may have a possible monitoring value for VNS activity. PMID- 17139391 TI - Myofasciitis and polyneuritis related to Buprenorphine abuse. AB - Various kinds of neuromuscular manifestations are known with the recreational drugs. We report an interesting case of extensive myositis and fasciitis of thigh following an injection of a solution of Buprenorphine. The inflammatory process affected the sciatic and obturator nerve as well. PMID- 17139392 TI - MRI in amyotrophic lateral sclerosis: hyperintensity of the corticospinal tract. AB - Amyotrophic lateral sclerosis is the most common form of motor neuron disease. The diagnosis is based on clinical and electromyography criteria. The primary role of imaging in amyotrophic lateral sclerosis is to exclude other causes such as cervical degenerative disk disease, Chiari malformation or multiple sclerosis. Imaging is also helpful in atypical cases of the disease. PMID- 17139393 TI - Ictal onset slow potential shifts recorded with hippocampal depth electrodes. AB - PURPOSE: Reports of direct current shifts at the onset of scalp-recorded seizures prompted us to inspect depth-recorded seizures for the presence of similar slow potential shifts at the onset of the seizure to determine whether slow potential (SP) shifts actually occur at the onset of depth-recorded seizures and if these shifts can facilitate localization of the seizure focus. METHODS: With the low frequency filter "opened" (LLF=0.1 Hz, HLF=70 Hz, 3 dB/octave), 32 seizures recorded with hippocampal depth and subdural electrodes were visually inspected to identify an SP shift at the onset of the seizure. A seizure was considered as having an SP shift when the slow potential waveform was > 1.5 sec in duration and > 100 microV in amplitude. Seizures were obtained from 5 subjects; 4 underwent epilepsy surgery (3=Engel I, 1=Engel II) and one received VNS. SP shift duration, peak voltage and polarity were measured for each seizure. The ability to identify seizures based on SP shift configuration was also evaluated. RESULTS: In 84% of the seizures, ictal onset was associated with a localized SP shift. Shift duration ranged from 1.5 sec to 11.5 sec (96% > 2 sec, 62% > 5 sec). The maximum shift ranged from 139 microV to 2305 microV (mean = 1123 microV, SD = 660 microV). In all the seizures, polarity was positive at the point of maximum shift. By visually examining the SP shift, seizures could be identified as originating from the same focus or from different foci. CONCLUSIONS: The onset of depth-recorded seizures appears to be commonly associated with a localized positive SP shift. An SP shift at the onset of depth-recorded seizures is likely to be a useful visual aid for localizing electrographic seizure onset. PMID- 17139394 TI - Toxicology today. PMID- 17139395 TI - Professionalism in the age of computerised medical records. AB - Electronic medical records have the potential to improve clinical care and to provide answers to important research questions. Research using existing medical records has provided important knowledge about the effectiveness and risks of widely-used medications. However, electronic medical records also raise ethical dilemmas regarding informed consent and confidentiality. Breaches of confidentiality with electronic records can be more severe than breaches with paper records. Furthermore, computerised health information raises new ethical dilemmas regarding direct advertisements of new drugs to patients, the impact of email on the doctor-patient relationship and the quality of outsourced radiology readings. Resolving these dilemmas may require new regulations and laws. In the interim, society will need to rely on physicians' professionalism to minimise the risks of electronic medical records and to ensure that the benefits outweigh the risks. PMID- 17139396 TI - Netting the evidence: finding pearls, not sewage. AB - The Internet is a powerful information resource that offers access to millions of files hosted on further millions of distributed computers. Unlike many information resources, however, the Internet focuses only on technical standards for information exchange. It does not address equally important concerns regarding the content and presentation of the information it displays. There is no editorial control, no unifying mechanism for maintenance or update, nor are there sanctions for the distribution of inaccurate and dangerous information. This overview seeks to guide the busy medical practitioner in their quest for high-quality research evidence. It stresses the value of key information management principles of focusing on the original question, filtering for high quality sources and finding evaluated information. It briefly summarises the advantages and limitations of three important sources of information: one-stop shops, general search engines and scholarly gateways. Throughout the article, key points are highlighted through the use of a realistic problem-based scenario. PMID- 17139397 TI - Characteristics of endosulfan poisoning: a study of 23 cases. AB - INTRODUCTION: Organochloride insecticides are chlorinated cyclic hydrocarbons having molecular weights in the range of 300-550 Da. Case series of endosulfan poisoning are extremely rare in the literature. We report 23 cases of endosulfan poisoning. METHODS: This retrospective study enrolled patients with endosulfan poisoning presenting to our emergency department from January to December 2005. The data were collected from clinical records and laboratory files. RESULTS: On admission, initial symptoms were nausea and vomiting in 17 patients (73.9 percent), seizures in five patients (21.7 percent), and dizziness in one patient (4.3 percent). Symptoms began within one hour after ingestion in 12 patients (52.2 percent), in the second hour in nine patients (39.1 percent), and in the third hour in two patients (8.7 percent). Seizure types were generalised tonic clonic in 16 patients (84.2 percent), and focal seizures in three patients (15.8 percent). 19 patients were observed for one day, two patients were observed for two days, and one patient was followed-up for ten days in the emergency department. One patient was transferred for liver transplantation on the fifth day to another centre. All patients were treated symptomatically by intravenous diazepam for controlling seizures. CONCLUSION: Endosulfan poisoning can be suspected in the presence of primary central nervous system manifestations including seizures, with or without clinical or laboratory evidence of other organ dysfunction such as liver failure. PMID- 17139398 TI - Experience with paraquat poisoning in a respiratory intensive care unit in North India. AB - INTRODUCTION: Paraquat poisoning is an uncommon entity in India. We report our experience of managing five patients with paraquat poisoning using immunosuppressive therapy. METHODS: Retrospective analysis of 84 patients admitted with a diagnosis of poisoning over the last eight years was performed. The data were presented in a descriptive fashion. RESULTS: Five (5.9 percent) out of the 84 patients were admitted with a diagnosis of paraquat poisoning. All patients were mechanically ventilated. All patients had hepatic failure with median peak bilirubin being 22.1 +/- 15.1 mg/dL (range 8.4-45.5). Four of the five patients had renal failure (median peak creatinine 3.8 +/- 1.5 mg/dL; range 3.4-11.1) requiring renal replacement therapy. All patients were treated with intravenous methylprednisolone 15 mg/kg/day for three consecutive days and intravenous cyclophosphamide 10 mg/kg/ day for two consecutive days, followed by intravenous dexamethasone 4 mg thrice a day until recovery or death. Two out of the five patients survived. Three died because of severe acute respiratory distress syndrome and multiorgan dysfunction syndrome. CONCLUSION: Paraquat poisoning is an uncommon entity in India, and is associated with a high mortality rate. There is a potential role for immunosuppressive therapy in patients with moderate to severe poisoning. PMID- 17139399 TI - Evaluation of anticarcinogenic effects of Clerodendron inerme on 7,12 dimethylbenz(a) anthracene-induced hamster buccal pouch carcinogenesis. AB - INTRODUCTION: Oral cancer is the fifth most frequent cancer worldwide and India has recorded the highest incidence (40-50 percent) of oral malignancy. Clerodendron inerme is used by Indian traditional practitioners for the treatment of various ailments, including cancer. Our aim was to investigate the chemopreventive potential of the aqueous leaf extract of Clerodendron inerme (CiAet) in 7,12-dimethylbenz(a) anthracene (DMBA)-induced hamster buccal pouch carcinogenesis. METHODS: We developed oral squamous cell carcinoma in the buccal pouch of male Syrian golden hamsters by painting them with 0.5% DMBA in liquid paraffin thrice a week for 14 weeks. The tumour incidence, tumour volume and tumour burden that were formed in the hamster buccal pouches were determined. RESULTS: Oral administration of CiAet at a dose of 500 mg/kg body weight to DMBA painted animals on days alternate to DMBA painting for 14 weeks significantly prevented the tumour incidence, tumour volume and tumour burden. CiAet also exerts potent antilipidperoxidative effect and improved the antioxidant defence system in DMBA-painted animals. The chemopreventive efficacy of CiAet was evident by inhibition of tumour formation (80%) in DMBA-painted animals. CONCLUSION: The chemopreventive potential of CiAet is probably due to its antilipidperoxidative effect or the presence of some potent bioactive chemopreventive principles in the leaves of Clerodendron inerme. PMID- 17139400 TI - A re-look at the duration of human pregnancy. AB - INTRODUCTION: The duration of human pregnancy is arbitrarily taken as 280 days (40 weeks). Foetuses are considered to be at high risk once pregnancy goes beyond the expected date of confinement. This study was carried out with the aim of determining the mean gestation age of low-risk pregnancies that went into spontaneous labour and the incidence of adverse outcomes in relation to gestation. METHODS: Low-risk singleton pregnancies admitted in spontaneous labour at a single community hospital in the Udupi district of Karnataka in South India, from December 2002 to December 2003, were analysed for mean gestational age at the onset of spontaneous labour and rates of perinatal complications by gestational age. RESULTS: Among the 1,094 women who went into spontaneous labour, the mean gestational age was 272.1 +/- 9 days. A significantly increased incidence of meconium-stained amniotic fluid beyond 39 weeks of gestation was observed. 783 of 1,094 women (80 percent) had delivered during the period of 261 280 days of pregnancy (period of one standard deviation around the mean gestational age at delivery). There was significant increase in perinatal morbidity indicators and mortality rates once the pregnancy carried beyond 280 days. CONCLUSION: Mean gestational age at the onset of labour for women native to the area of study was 272 days (standard deviation 9 days). Pregnancies beyond a duration of 280 days showed significantly increased perinatal morbidity. It is suggested that there is a need for determining the length of gestation and to compile gestation-wise incidence of meconium-stained amniotic fluid as an indicator of foetal maturity or the undisclosed risk factor, in addition to other neonatal morbidity indicators for different populations. PMID- 17139401 TI - Clinical profile, electrodiagnosis and outcome in patients with carpal tunnel syndrome: a Singapore perspective. AB - INTRODUCTION: Carpal tunnel syndrome (CTS ) is the most common entrapment neuropathy seen in our neurodiagnostic laboratory referrals. We describe the clinical profile, and outcome in patients with electrophysiological diagnosis of CTS seen in our centre over a six month period. METHODS: A retrospective study was carried out and included 134 consecutive patients with CTS referred to the Neurodiagnostic Laboratory, National Neuroscience Institute, from October 2003 to March 2004, for the confirmatory testing. Severity grade was assigned following American Association of Electrodiagnostic Medicine criteria of CTS. RESULTS: The majority of patients were female (81.3 percent) with mean age of presentation being 53.6 years. Chinese women constitute the majority racial group. Paraesthesia (70.1 percent) and numbness (19.4 percent) were the presenting sensory symptoms. In the nerve conduction study, 108 patients had bilateral CTS with 35 having unilateral symptoms. Dominant hand involvement was present in 92.3 percent. Overall, 40.3 percent had mild, 46.3 percent had moderate and 13.4 percent had severe CTS, with median duration of symptoms of two, four and 12 months, respectively. Follow-up data were available for 115 patients. 27 patients with surgical treatment showed resolution or improvement in 53.3 percent with moderate CTS, and 83.3 percent with severe CTS, at three-month follow-up. 14 patients turned up for six-month follow-up and 92.9 percent showed improvement in symptoms. 88 patients were managed conservatively; symptoms were unchanged or worsened in 80.6 percent with mild CTS, 65.9 percent with moderate CTS, and 62.5 percent with severe CTS at three-month follow-up. Of the 54 patients who turned up for six-month follow-up, the clinical symptom remain unchanged or worsened in 68.5 percent. CONCLUSION: The severity of CTS is associated with longer duration of symptoms. Sensory symptoms and dominant hand involvement is more common. There is a high default rate in the clinical follow-up. Early surgical intervention results in either resolution or improvement in symptoms, whereas conservative management does not affect the natural history with symptoms that persisted or worsened with time. PMID- 17139402 TI - The product of outcome-based undergraduate medical education: competencies and readiness for internship. AB - INTRODUCTION: Outcome-based curriculum is adopted at the International Medical University (IMU), Malaysia, where specific learning objectives are laid out progressively under eight major outcomes. We present an outcome-guided, self reported competency profile of our undergraduate students near the end of their training, focusing on elements that are considered most immediately relevant for their internship. METHODS: Anonymous surveys were conducted on two cohorts of medical students in their final semester at IMU. The surveys covered a range of competencies, including practical skills, ward routines, generic attributes and evidence-based medicine, grouped under the exit outcomes as defined by the university. RESULTS: A total of 92 students were assessed. In general, the students were confident of their ability on common practical skills and ward routines. They were comfortable with the level of professionalism and personal attributes required for internship, with the prospect of handling unexpected additional tasks and working away from home perceived as the main difficulties. Most students referred to at least three sources of clinical information to answer their clinical queries. However, they referred more to single journals than databases or collections. The majority could critically appraise journal articles to a variable extent, but nearly half took 30 minutes or longer to trace an abstract of interest. CONCLUSION: This report demonstrates the strength of outcome-based curriculum in its ability to produce competent students that are well prepared for their internship. Assessing students using this educational approach provides a clear picture of their strengths and weaknesses, and identifies stages in their training where additional inputs are required. PMID- 17139403 TI - Solitary rectal ulcer syndrome: characteristics, outcomes and predictive profiles for persistent bleeding per rectum. AB - INTRODUCTION: Solitary rectal ulcer syndrome (SRUS) is a rare disorder of defaecation and persistence of symptoms is common, particularly bleeding per rectum (BPR). This study assessed the clinical, endoscopical characteristics and predictive profiles for persistent BPR. METHODS: 28 patients (14 males) with biopsy-proven SRUS were identified from 1989 to 2003, and the clinical and endoscopical findings were retrospectively reviewed. RESULTS: At presentation, the mean age was 29.5 +/- 16.1 (range ten to 81) years. Common symptoms reported included BPR (86 percent), abdominal pain (36 percent), mucus per rectum (25 percent), straining at defaecation (25 percent), diarrhoea (14 percent) and constipation (14 percent). Digital manual evacuation was reported by 11 percent. 68 percent were anaemic and 57 percent required blood transfusion. Lesions were located anteriorly (38.5 percent), posteriorly (30.7 percent) and circumferentially (31.8 percent). The lesions were multiple (34 percent), ulcerative (64.3 percent) and polypoidal/nodular (32.1 percent). At a mean follow up of 43.5 +/- 36 months, 64 percent (n=18) had multiple admissions (mean 3.1, range one to 12), mainly for transfusion (mean 7.4 units, range two to 27). There was no difference in clinical responses between patients with polypoidal/nodular or ulcerative lesions (p-value is 0.653). Follow-up endoscopies showed improvement (58 percent), progression (21 percent) and no change (21 percent) in the lesions. Four patients had surgery for concerns of neoplasms (n=2) and persistent BPR (n=2). BPR was persistent in 39 percent. Presence of abdominal pain (p-value is 0.008) and passage of abnormal stool (p-value is 0.002) were predictive of persistent BPR. CONCLUSION: SRUS occurs predominantly in young patients and despite being a benign condition, morbidity remains a problem. Patients' profiles are predictive of persistent BPR. PMID- 17139404 TI - AIDS-associated Kaposi's sarcoma in Northeastern Nigeria. AB - INTRODUCTION: Kaposi's sarcoma is an acquired immunodeficiency syndrome (AIDS) defining illness, and with the size of the human immunodeficiency virus (HIV)/AIDS pandemic in sub-Saharan Africa, AIDS-related Kaposi's sarcoma (KS) are now being diagnosed more frequently, although the true incidence of HIV associated KS is not known. The clinical presentations of AIDS-related KS varied markedly across the African continent. This article reports a series of unusual clinical presentations of the tumour in Northeastern Nigeria. METHODS: This is a prospective study carried out from September 2003 to August 2005, at the University of Maiduguri Teaching Hospital, Borno State, Northeastern Nigeria. RESULTS: 20 cases of histologically-confirmed KS were prospectively studied. There were 17 (85 percent) men and three (15 percent) women, giving a male to female ratio of 5.7:1. Their ages ranged from 21-45 (median 37) years. 18 (90 percent) of the patients were anaemic. Mean haematocrit value, CD4+ cell count and duration of symptoms for all the patients were 29.5 +/- 7.5 percent, 119.0 +/ 91.4 cells per microlitre and 3.5 +/-1.7 months, respectively. Multiple lesions were a common presentation affecting sites such as lower limbs, trunk, conjunctiva, upper limbs and rectum as well as penis, lymph node, scrotum and oropharynx. CONCLUSION: Contrary to other reports that KS is not associated with HIV infection, our study has demonstrated otherwise. This study also showed that both sexes are affected but with a male preponderance. KS is also a late presentation of the HIV/AIDS disease spectrum in our environment and has varied clinical manifestations. There is an urgent need to develop health education programmes to enhance the understanding of this disease and how it spreads, particularly among the young generation. PMID- 17139405 TI - Clinical spectrum of Parkinson's disease from Pakistan. AB - INTRODUCTION: Parkinson's disease is an idiopathic disorder of the extrapyramidal system. It has a worldwide prevalence but data from developing countries is scanty. We describe the clinical spectrum of the disease from Pakistan, a developing country. METHODS: Patients with Parkinson's disease, over a period of 11 years, were identified by ICD-9 coding system of the hospital medical records. Demographical characteristics, clinical features, laboratory investigations and radiological investigations were recorded and analysed. RESULTS: A total of 80 patients were identified. 50 (63 percent) were males and 30 (37 percent) were females. Mean age of onset of the disease was 54 years. 47 (59 percent) patients had onset of illness during the sixth or seventh decade of life. Mean duration of illness at the time of presentation was five years. Rigidity, bradykinesia, tremors, hypomimia, primitive reflexes, difficulty in performing fine work and walking difficulty were the most common clinical features. 52 (65 percent) patients had stage I or II (Hoehn-Yahr staging) disease at the time of presentation. 56 (70 percent) patients had predominantly unilateral symptoms. 15 (19 percent) patients had cognitive impairment. Cognitive decline was more common in the elderly and in patients with disease duration of longer than ten years. CONCLUSION: Parkinson's disease is more common in males. Tremor, rigidity, walking difficulty, bradykinesia and difficulty in performing fine work are the commonest clinical features. Disease severity increases with duration of the disease. Cognitive impairment is not uncommon in these patients and is associated with disease duration and age of onset of the illness. PMID- 17139406 TI - Ecthyma gangrenosum: a manifestation of Pseudomonas sepsis in three paediatric patients. AB - Pseudomonas aeruginosa sepsis rarely occurs in healthy children. In immunocompromised children, it usually carries a high mortality rate. Ecthyma gangrenosum is a known cutaneous manifestation of Pseudomonas septicaemia. Three paediatric cases of Pseudomonas aeruginosa septicaemia with ecthyma gangrenosum were retrospectively reviewed. The three patients were aged seven years, seven months, and five months, respectively. An underlying disease of hypogammaglobulinaemia was present in the oldest patient. Blood cultures grew Pseudomonas aeruginosa in all three patients. All underwent repeated wound debridement and received intravenous ceftazidime and an aminoglycoside for a minimum of two weeks. One needed colostomy and subsequent posterior sagittal anorectoplasty as a result of complete obliteration of the anal canal from the ecthyma. There was no mortality. In conclusion, Pseudomonas aeruginosa sepsis should be treated early. Recognition of ecthyma gangrenosum as a manifestation of this problem can allow early institution of the appropriate antibiotics before culture results. PMID- 17139407 TI - Massive pre-placental and subchorionic haematoma. AB - We report an unusual case of massive pre-placental and subchorionic haematoma occurring in a 26-year-old woman who presented with antepartum haemorrhage at 24 weeks gestation. Ultrasonography showed a subchorionic haematoma in the lower posterior uterine wall measuring 5.0 cm in largest diameter. There was also a separate irregular multiloculated structure measuring 4.3 cm in largest diameter on the surface of the placenta, due to a pre-placental haematoma. The subchorionic haematoma diminished in size over time, while the pre-placental haematoma continued to grow, measuring 9.0 cm at 28 weeks, and 9.3 cm at 32 weeks. At 32 weeks, the patient presented with premature rupture of membranes and four days later, an emergency caesarean section was performed when the patient had another episode of severe antepartum haemorrhage. Both mother and child recovered well. The current literature on such haematomas is reviewed. PMID- 17139408 TI - Cantrell syndrome in one of a set of monozygotic twins. AB - Cantrell syndrome is a rare syndrome of congenital defects involving the abdominal wall, sternum, diaphragm, pericardium and the heart. The spectrum of anomalies varies widely. A full syndrome, probable syndrome and incomplete expression have been described. Less than 160 cases have been described in the world literature. Only one set of monozygotic twins concordant with the syndrome has been reported. To our knowledge, the syndrome affecting only one of a set of twins has not been reported. We report a premature infant, the first-born twin, with the syndrome. The omphalocoele was large, and heart abnormalities included dextrocardia with atrial septal defect. The co-twin is normal. PMID- 17139409 TI - Subdural empyema post-chemoradiotherapy for nasopharyngeal carcinoma. AB - Nasopharyngeal carcinoma is a common malignancy in the Asian Chinese population. The first-line treatment for Stage III/IV disease has in recent times shifted from radiotherapy to that of concurrent chemoradiotherapy. The treatment is not infrequently associated with in-field complications. We describe a rare case of one such complication -- subdural empyema developing post-chemoradiotherapy in a 56-year-old man with Stage IVB nasopharyngeal carcinoma. PMID- 17139410 TI - Is this a variant of Lemierre's syndrome? AB - Lemierre's syndrome is an oropharyngeal infection leading to secondary septic thrombophlebitis of the internal jugular vein (IJV). It is classically anaerobic in origin and Fusobacterium necrophorum is the commonest pathogen. We report an unusual variant of this condition in a 68-year-man with uncontrolled diabetes mellitus who presented with extensive cellulitis of the left neck. No primary oropharyngeal sepsis was detected. Neck radiographs revealed soft tissue gas and computed tomography confirmed neck space infection with gas formation and complete thrombosis of the left IJV. Klebsiella pneumoniae was isolated from the fluid, tissue and blood samples. Repeated debridement was required until complete healing. Computed tomography of the neck at six months showed resolution of infection, although the vein remained thrombosed. Although the radiological features resemble Lemierre's syndrome, this case differs from it in the absence of oropharyngeal sepsis and presence of background diabetes mellitus. The isolation of Klebsiella pneumoniae as a cause has not been previously described. PMID- 17139411 TI - Dispelling the myths of epidural pain relief in childbirth. PMID- 17139418 TI - Description of a new dinoflagellate with a diatom endosymbiont, Durinskia capensis sp. nov. (Peridiniales, Dinophyceae) from South Africa. AB - A new dinoflagellate Durinskia capensis Pienaar, Sakai et Horiguchi sp. nov. (Peridiniales, Dinophyceae), from tidal pools along the west coast of the Cape Peninsula, Republic of South Africa, is described. The dinoflagellate produces characteristic dense orange-red colored blooms in tidal pools. The organism is characterized by having a eukaryotic endosymbiotic alga. Ultrastructure study revealed the organism has a cellular construction similar to that of other diatom harboring dinoflagellates. The cell is thecate and the plate formula is: Po, x, 4', 2a, 6'', 5c, 4s, 5''', 2'''', which is the same as that of Durinskia baltica, the type species of the genus Durinskia. D. capensis can, however, be distinguished from D. baltica by overall cell shape, the relative size of the 1a and 2a plates, the degree of cingular displacement, and the shape of the eyespot. Our molecular analysis based on SSU rDNA revealed that D. capensis is closely allied to D. baltica, thus supporting the assignment of this new species to this genus. This Durinskia clade takes a sister position to another diatom-harboring dinoflagellate clade, which includes Kryptoperidinium foliaceum and Galeidinium rugatum. Molecular analysis based on the rbcL gene sequence and ultrastructure study revealed that the endosymbiont of D. capensis is a diatom. The SSU rDNA gene trees indicated that four species with a diatom endosymbiont formed a clade, suggesting a single endosymbiotic origin. PMID- 17139419 TI - Development of genotype-independent regeneration system for transformation of rice (Oryza sativa ssp. indica). AB - Rice (Oryza sativa ssp. indica) is an important economic crop in many countries. Although a variety of conventional methods have been developed to improve this plant, manipulation by genetic engineering is still complicated. We have established a system of multiple shoot regeneration from rice shoot apical meristem. By use of MS medium containing 4 mg L(-1) thidiazuron (TDZ) multiple shoots were successfully developed directly from the meristem without an intervening callus stage. All rice cultivars tested responded well on the medium and regenerated to plantlets that were readily transferred to soil within 5-8 weeks. The tissue culture system was suitable for Agrobacterium-mediated transformation and different factors affecting transformation efficiency were investigated. Agrobacterium strain EHA105 containing the plasmid pCAMBIA1301 was used. The lowest concentration of hygromycin B in combined with either 250 mg L( 1) carbenicillin or 250 mg L(-1) cefotaxime to kill the rice shoot apical meristem was 50 mg L(-1) and carbenicillin was more effective than cefotaxime. Two-hundred micromolar acetosyringone had no effect on the efficiency of transient expression. Sonication of rice shoot apical meristem for 10 s during bacterial immersion increased transient GUS expression in three-day co-cultivated seedlings. The gus gene was found to be integrated into the genome of the T(0) transformant plantlets. PMID- 17139420 TI - Developmental processes of achlorophyllous orchid, Epipogium roseum: from seed germination to flowering under symbiotic cultivation with mycorrhizal fungus. AB - We have achieved the symbiotic cultivation of an apparently achlorophyllous orchid, Epipogium roseum Lindl., with a mycorrhizal fungus isolated from an underground organ of this orchid. Although the seed germination rate was extremely low, subsequent growth from protocorm to flowering was induced in a medium containing volcanic soils and sawdust. Stolons elongated from each protocorm, and rhizomes were formed at certain intervals on the stolons. Some of the rhizomes developed into a coralloid form, and tubers were formed from the coralloid rhizomes. The coralloid rhizomes degenerated concurrently with maturation of the tubers. Six months after seed sowing, around 80 tubers were produced from a single protocorm. An inflorescence appeared from each of the large tubers, and the process to flowering was observed in one of these. Consequently, the developmental processes from seed to flowering in E. roseum was clearly revealed in this study. PMID- 17139421 TI - Biliary complications after deceased-donor orthotopic liver transplantation. AB - A wide range of potential biliary complications can occur after orthotopic liver transplantation (OLT). The most common biliary complications are bile leaks, anastomotic and intrahepatic strictures, stones, and ampullary dyfunction, which may occur in up to 20%-40% of OLT recipients. Leaks predominate in the early posttransplant period; stricture formation typically develops gradually over time. However, with the advent of new techniques, such as split-liver, reduced size, and living-donor liver transplantation, the spectrum of biliary complications has changed. Risk factors for biliary complications comprise technical failure; T-tube or stent-related complications; hepatic artery thrombosis; bleeding; ischemia/reperfusion injury; and other immunological, nonimmunological, and infectious complications. Noninvasive diagnostic methods have been established and treatment modalities have been modified towards a primarily nonoperative, endoscopy-based strategy. Besides, the management of biliary complications after OLT requires a multidisciplinary approach, in which interventional and endoscopic treatment options have to be weighed up against surgical treatment options. The etiology and spectrum of bile duct complications, their diagnosis, and their treatment will be reviewed in this article. PMID- 17139422 TI - Biliary complications in recipients of living-donor liver transplantation. AB - The key points of the management of biliary complications in recipients of living donor liver transplantation are described. The characteristics of these complications are somewhat different from those in deceased-donor liver transplantation, mainly due to the technical difficulties. Appropriate prevention, diagnosis, and treatment are essential for successful transplants, to avoid the development of secondary biliary cirrhosis when complication occurs. PMID- 17139423 TI - Endoscopic treatment of biliary complications after right-lobe living-donor liver transplantation with duct-to-duct biliary anastomosis. AB - BACKGROUND/PURPOSE: The aims of this study were to characterize the features of the biliary complications that occur after right-lobe living-donor liver transplantation (RL-LDLT) with duct-to-duct biliary anastomosis, and to evaluate the efficacy of treating biliary complications endoscopically. METHODS: The records of 273 consecutive patients who underwent RL-LDLT with duct-to-duct biliary anastomosis from July 1999 through July 2005 at Kyoto University Hospital were reviewed to determine the overall incidence of postoperative biliary complications and the outcome of endoscopic repair of those complications. RESULTS: Biliary complications occurred in 93 (34.1%) of the patients. These complications were: 80 biliary strictures (75 anastomotic and 5 nonanastomotic) and 16 biliary leakages (5 patients with biliary leakage also had a biliary stricture); most (72%) of the anastomotic strictures were complex (i.e., fork shaped or trident-shaped). The strictures and leakages were repaired by the endoscopic placement of multiple inside stents above the sphincter of Oddi, and by nasobiliary drainage, respectively. The procedure was successful in repairing 51 (68.0%) of the anastomotic strictures and 8 (50.0%) of the biliary leakages. CONCLUSIONS: Endoscopic stenting of the bile ducts is efficacious in treating biliary complications related to RL-LDLT with duct-to-duct biliary anastomosis and the stenting should be attempted before surgical revision of strictures and leakages. PMID- 17139424 TI - Intrahepatic biliary strictures after liver transplantation. AB - Biliary complication has been one of the most common complications after liver transplantation. Nonanastomotic strictures and dilatations involving the intrahepatic biliary tree have been recognized as biliary complications. These lesions were reported to be associated with hepatic artery thrombosis; prolonged preservation time; ABO-incompatible organs; and immunological injury, including injuries to vascular endothelial cells (chronic rejection) and the bile duct (primary sclerosing cholangitis). However, the etiology of these lesions appeared to be mostly related to ischemic injury. Anatomical research on the arterial supply of the bile duct has provided further insights into bile duct blood supply and its surgical implications. The biliary tract is supplied with arterial blood by a vasculature called the peribiliary vascular plexus. Any injury to the peribiliary vascular plexus may contribute to ischemic death of the biliary system mucosa. At many points, the process of liver transplantation exposes the endothelial cells and peribiliary vascular plexus to ischemic injury. The majority of intrahepatic biliary strictures (IHBS) are diffuse or bilateral. A percutaneous or an endoscopic approach has been used as the initial treatment. However, a low threshold for surgical intervention (retransplantation) should be adopted, because these patients demonstrate high mortality. The aim of this article is to review the anatomy, etiology, clinical picture, diagnosis, management, and prognosis of IHBS after liver transplantation. PMID- 17139425 TI - Causes and consequences of ischemic-type biliary lesions after liver transplantation. AB - Biliary complications are a major source of morbidity, graft loss, and even mortality after liver transplantation. The most troublesome are the so-called ischemic-type biliary lesions (ITBL), with an incidence varying between 5% and 15%. ITBL is a radiological diagnosis, characterized by intrahepatic strictures and dilatations on a cholangiogram, in the absence of hepatic artery thrombosis. Several risk factors for ITBL have been identified, strongly suggesting a multifactorial origin. The main categories of risk factors for ITBL include ischemia-related injury; immunologically induced injury; and cytotoxic injury, induced by bile salts. However, in many cases no specific risk factor can be identified. Ischemia-related injury comprises prolonged ischemic times and disturbance in blood flow through the peribiliary vascular plexus. Immunological injury is assumed to be a risk factor based on the relationship of ITBL with ABO incompatibility, polymorphism in genes coding for chemokines, and pre-existing immunologically mediated diseases such as primary sclerosing cholangitis and autoimmune hepatitis. The clinical presentation of patients with ITBL is often not specific; symptoms may include fever, abdominal complaints, and increased cholestasis on liver function tests. Diagnosis is made by imaging studies of the bile ducts. Treatment starts with relieving the symptoms of cholestasis and dilatation by endoscopic retrograde cholangiopancreaticography (ERCP) or percutaneous transhepatic cholangiodrainage (PTCD), followed by stenting if possible. Eventually up to 50% of the patients with ITBL will require a retransplantation or may die. In selected patients, a retransplantation can be avoided or delayed by resection of the extra-hepatic bile ducts and construction of a hepaticojejunostomy. More research on the pathogenesis of ITBL is needed before more specific preventive or therapeutic strategies can be developed. PMID- 17139426 TI - Ex vivo extended left hepatectomy with caval preservation, temporary portacaval shunt, and reconstruction of the right hepatic vein outflow using a reversed portal vein bifurcation graft. AB - Liver resections that require ex vivo techniques occur rarely, but when done are generally performed on veno-veno bypass to maintain venous return and decompress the portal circulation during the anhepatic phase of the procedure. We describe an ex vivo extended left hepatectomy that was performed with preservation of the inferior vena cava and the use of a temporary portacaval shunt to eliminate the need for veno-venous bypass. Ex vivo resection allowed reconstruction of right hepatic vein branches, using the patient's reversed portal vein bifurcation as a graft to provide venous outflow. PMID- 17139427 TI - Retrospective analysis of outcome in 63 gallbladder carcinoma patients after radical resection. AB - BACKGROUND/PURPOSE: The aim of this study was to evaluate factors influencing outcome in gallbladder carcinoma after radical resection, in order to identify those patients benefiting from radical surgery. METHODS: Sixty-three patients (13 pT1, 28 pT2, 14 pT3, and 8 pT4) who underwent surgical resection for gallbladder carcinoma were retrospectively reviewed. Correlations were sought between survival and factors such as the depth of invasion (pT) including the status of infiltration of the hepatoduodenal ligament (pBinf) and liver bed (pHinf), the extent of lymph node metastasis, and other pathologic factors. RESULTS: Multivariate analysis showed that pBinf, pHinf, and lymph node metastasis were significant prognostic factors. We also analyzed survival rates for each operative procedure. There was no difference in survival between patients with or without bile duct resection for lymph node metastasis. The 5-year survival rates of pHinf-negative patients with stage 1B or more advanced disease after gallbladder bed resection or bisegmentectomy 4a,5 versus those without liver resection were 66% and 0%, respectively. Twelve patients survived for more than 5 years after surgery, including one patient undergoing HPD (liver resection with pancreatoduodenectomy) with positive lymph node metastasis; none was pBinf positive. CONCLUSIONS: Several factyors were identified as having prognostic significance for survival in patients with gallbladder carcinomas, and we suggest that radical surgery may be indicated for selected patients with advanced disease. PMID- 17139428 TI - Combined hepatocellular and cholangiocarcinoma: clinical features and prognostic study in a Thai population. AB - BACKGROUND/PURPOSE: Combined hepatocellular and cholangiocarcinoma (HCC-CC) is an uncommon subtype of primary liver cancer, the clinicopathological features of which have rarely been reported in detail. Some authors believe that HCC-CC behaves like HCC, but biliary differentiation may be associated with poorer prognosis. Moreover, CC has more frequent lymph node metastases. In this study, we aimed to determine the clinical course and survival outcome of HCC-CC patients in a Thai population by comparing them with patients with ordinary HCC. METHODS: The clinicopathological features of patients who were diagnosed with HCC-CC at Ramathibodi Hospital during 2000-2004 were retrospectively studied by comparing them with the features of patients suffering from ordinary HCC. Twenty-five patients who were diagnosed with HCC-CC were included in this study, and subsequently 50 patients with HCC who had tissues taken during the same period were selected randomly from among 148 HCC patients. Statistical analysis was done by using SPSS version 10.0. The Kaplan-Meier method was used to assess the survival rate. Multiple logistic regression analysis was performed to assess correlations. A value of P < 0.05 was considered statistically significant. RESULTS: There were no significant differences in etiologic risk factors between HCC-CC and HCC patients: cirrhosis (50% vs 44%), chronic alcohol abuse (36% vs 43%), presence of hepatitis B surface antigen (HBsAg; 66% vs 78%) and presence of hepatitis C virus (HCV) antibody (13% vs 3%). The serum alpha-fetoprotein (AFP) value in the HCC-CC group was lower than that in the HCC group (5.87 vs 41.46 ng/ml). No differences in tumor characteristics or liver status (tumor size, presence of multinodular lesions, portal vein thrombosis, intrahepatic bile duct dilatation, intraabdominal lymphadenopathy, extrahepatic metastasis, liver cirrhosis, portal hypertension, and ascites) between these two groups were found. The overall median survival of HCC-CC patients was 38 weeks while that of HCC patients was 54 weeks. Multivariate analysis showed that elevated carbohydrate antigen (CA)19-9 (>/=80 U/ml) and the presence of intrahepatic bile duct dilatation were independent risk factors for worse survival. CONCLUSIONS: The demographic and clinical features of patients with combined HCC-CC were similar to those of patients with HCC. The presence of cholangiocellular differentiation appeared to worsen the prognosis when compared with pure HCC, although this difference did not reach statistical significance. An increased CA19-9 level and intrahepatic bile duct dilatation in patients with HCC-CC were considered to be independent factors that suggested poor prognosis. PMID- 17139429 TI - Early detection of liver failure after hepatectomy by indocyanine green elimination rate measured by pulse dye-densitometry. AB - BACKGROUND/PURPOSE: Following a major hepatectomy, some degree of clinical and biochemical dysfunction occurs. Surgeons usually check serum total bilirubin levels to diagnose postoperative liver dysfunction. However, we cannot predict liver failure by biochemical data alone within the early postoperative period. Using newly developed pulse dye-densitometry (PDD), we measured serial postoperative indocyanine green elimination rate (ICG-K) values and investigated the possible relation between postoperative ICG-K values and complications. METHODS: Fifty-one patients scheduled for hepatectomy between January 2000 and December 2002 were enrolled. Pulse-dye densitometry was used to evaluate postoperative liver function. We analyzed the relation between postoperative ICG K and postoperative outcome, assessed in terms of morbidity and mortality. RESULTS: Liver failure was seen in seven patients. The ICG-K value on postoperative day 1 in patients with liver failure was significantly lower than that in patients without liver failure (0.070 +/- 0.018 vs 0.152 +/- 0.056/min respectively; P < 0.001). There were no differences between preoperative ICG-K values in patients with and without liver failure. The sensitivity and specificity of an ICG-K value of less than 0.07 on postoperative day 1 were 71.4% and 95.5%, respectively, for predicting liver failure. CONCLUSIONS: We can measure the ICG-K value by PDD at the bedside without time delay, and we can predict liver failure in the early postoperative period by the ICG-K values on postoperative day 1. ICG-K values measured by PDD can provide important information for perioperative management. PMID- 17139430 TI - Nodal involvement as an indicator of postoperative liver metastasis in carcinoma of the papilla of Vater. AB - BACKGROUND/PURPOSE: Although lymph node metastatic involvement is one of the most important prognostic factors for carcinoma of the papilla of Vater, a detailed analysis of this factor in relation to prognosis has not been conducted. METHODS: From 1985 to 2003, 29 patients with carcinoma of the papilla of Vater underwent pancreaticoduodenectomy and dissection of regional lymph nodes at Yamagata University Hospital. We analyzed clinicopathologic variables in relation to prognosis and precisely evaluated nodal involvement in each patient to determine lymphatic flow. Furthermore, the relationship between recurrent site and nodal involvement was investigated. RESULTS: The overall survival rate was 55% at 5 years. The significant prognostic factors were morphological ulcer formation (P = 0.04), histological type (P = 0.03), nodal involvement (P = 0.002), and lymphatic invasion (P = 0.03). Multivariate analysis indicated no independent factor, but nodal involvement may be the strongest prognostic factor. The overall rate of nodal involvement was 41.4% (12 of 29 patients). The metastatic rates in the superior posterior pancreaticoduodenal lymph nodes, the inferior posterior pancreaticoduodenal lymph nodes, the superior mesenteric lymph nodes, and paraaortic lymph nodes were high (31.0%, 20.7%, 17.2%, and 13.8%, respectively). Patients with nodal involvement had a significantly higher rate of liver metastasis after surgery than those without it (P = 0.02). Ulcer formation and histological type were significantly correlated with nodal involvement (P = 0.05 and P = 0.002, respectively). CONCLUSIONS: Nodal involvement is the most important prognostic factor in patients with carcinoma of the papilla of Vater. Patients with nodal involvement are at high risk of liver metastasis; therefore, adjuvant therapy may be necessary for the control of liver metastasis. Preoperative ulcer formation and histological type in the biopsy specimen are good indicators for extended lymph node dissection and adjuvant therapy, because these variables are correlated with nodal involvement. However, our data revealed only the sites of the positive nodes, without addressing the effect of extended lymph node dissection and adjuvant chemotherapy. To date, there has been reporting of extended lymph node dissection and adjuvant chemotherapy in patients with carcinoma of the papilla of Vater. Further studies will be necessary to resolve these problems. PMID- 17139431 TI - Histopathological study of invasive ductal carcinoma (IDC) of the pancreas without associated cancerous occlusion of the main pancreatic duct. AB - BACKGROUND/PURPOSE: [corrected] Invasive ductal carcinoma (IDC) of the pancreas may be associated with cancerous occlusion of the main pancreatic duct (MPD) in its growth process, but at quite low a frequency; there are patients who do not develop this occlusion. METHODS: This study examined the histological features of surgical specimens from 8 patients with IDC without MPD occlusion, in comparison to 32 patients with IDC with this occlusion (controls). The pancreatic duct was identified by confirming the presence of mural elastic fibers on the wall of the pancreatic duct. Immunohistochemical staining was done with Ki-67 antibody. RESULTS: The frequency of IDC without MPD occlusion was very low (5.0% [2/40] patients at Kurume University and 3.1% [4/126] patients at Juntendo University). The number of intraductal carcinoma components was 1.5 +/- 1.1 per specimen in the IDCs without occlusion and 5.9 +/- 2.4 in the controls (P < 0.001). The Ki-67 labeling index was 18.0 +/- 11.7% in the IDCs without occlusion and 30.0 +/- 12.1% in the controls (P < 0.05). The number of intraductal carcinoma components and the Ki-67 labeling index were significantly lower in the IDCs without occlusion than in the controls. CONCLUSIONS: Our findings suggested that these two types of IDC could have different biological features. PMID- 17139432 TI - Gallbladder cancer: surgical results after cholecystectomy in 25 patients with lamina propria invasion and 26 patients with muscular layer invasion. AB - BACKGROUND/PURPOSE: It has been stated that simple cholecystectomy is sufficient treatment for all patients with pT1 gallbladder cancer. However, other authors note the necessity of carrying out extended surgery when there is muscular-layer involvement. METHODS: A consecutive series of gallbladder carcinomas with lamina propria or muscular layer invasion were analyzed. Between July 1982 and December 2000, 51 patients with pT1 gallbladder carcinomas were treated with simple cholecystectomy (group A, 25 patients with lamina propria-invasion; group B, 26 patients with muscular-layer invasion). Patients with intraepithelial carcinomas were excluded from the study. RESULTS: There were no differences between the groups in average age, sex ratios, association with other tumors, histologic type, malignancy grade, cholecystitis type, macroscopic aspects, lymph node status, or treatment applied. After an average of 6 years' follow-up, no patients in group A and nine patients (34.6%) in group B died due to gallbladder carcinoma. Cystic lymph nodes could be studied in five of these nine patients who relapsed, and the results were negative for metastasis. Lymphatic or venous invasion was observed in five of these nine patients. CONCLUSIONS: According to these results, cholecystectomy is not sufficient treatment for gallbladder carcinoma with muscular-layer invasion. PMID- 17139433 TI - The effects of isothermic and hypothermic carbon dioxide pneumoperitoneum on respiratory function test results. AB - BACKGROUND/PURPOSE: This study investigated the effects of isothermic and hypothermic carbon dioxide, used for pneumoperitoneum during laparoscopic cholecystectomy, on respiratory function test results. METHODS: Thirty patients who underwent elective laparoscopic cholecystectomy were enrolled in this prospective randomized study. The patients were divided into two groups. Carbon dioxide at 37 degrees C (isothermic) was used in the isothermic group, and carbon dioxide at 21 degrees C (hypothermic) was used in the hypothermic group. Respiratory function tests were performed in the preoperative period and at 12 h after the operation. RESULTS: Mean forced vital capacity (FVC), forced expiratory volume (FEV1), maximum peak expiratory flow (PEF), and the FEV1/FVC ratio were significantly higher in the isothermic group than in the hypothermic group (P < 0.05). CONCLUSIONS: Using isothermic carbon dioxide for pneumoperitoneum has fewer negative effects than hypothermic carbon dioxide on respiratory function tests results. Isothermic carbon dioxide may be preferable for patients with respiratory problems. PMID- 17139434 TI - Synchronously resected double primary hepatic cancers - hepatocellular carcinoma and cholangiolocellular carcinoma. AB - The frequency of double primary cancers in the liver is very low. All reported cases are double cancers consisting of hepatocellular carcinoma (HCC) and intrahepatic cholangiocellular carcinoma (CCC). We herein report a surgical patient who had simultaneous double cancers consisting of HCC and cholangiolocellular carcinoma (CoCC). This is the first case report of such a patient. A 70-year-old Japanese man was admitted to our hospital for further examination of two hepatic nodules. He had a history of schistosomiasis japonica, idiopathic pulmonary fibrosis, and diabetes mellitus. Laboratory data revealed that hepatitis C virus (HCV) antibody was positive and hepatic enzymes were slightly elevated. The level of prothrombin induced by vitamin K absence or antagonist II was elevated. Computed tomography depicted two tumors; one, measuring 4.0 cm in diameter, was in the medial segment and the other, 2.2 cm in diameter, was in the posterior superior segment of the liver. The larger tumor showed contrast enhancement and the smaller one showed enhancement at the tumor periphery in the hepatic arterial phase. In the portal phase, the larger tumor became less dense than the liver parenchyma, but the periphery of the smaller one showed continuous enhancement. He underwent an operation under a diagnosis of double hepatic cancers, consisting of HCC and CCC. However, microscopic examination of the resected tumors revealed that the larger tumor was moderately differentiated HCC and the smaller one was CoCC. PMID- 17139435 TI - Extracystic biliary carcinoma associated with anomalous pancreaticobiliary junction and cysts. AB - Anomalous pancreaticobiliary junction with cystic dilatation of the biliary tract is usually associated with carcinoma arising from the cyst wall. We report an extracystic location of biliary carcinoma in the presence of anomalous pancreaticobiliary junction and cysts in a patient with obstruction of the origin of the left hepatic duct who underwent hepatobiliary resection. Cholangiocarcinoma was found to have arisen in a noncystic left hepatic duct, in conjunction with cystic dilatation involving both the cystic and common bile ducts. The present case supports a relationship between anomalous pancreaticobiliary junction and biliary carcinogenesis that may affect the extracystic biliary tree. PMID- 17139436 TI - Porcelain gallbladder complicated with pancreas divisum. AB - We report a rare case of porcelain gallbladder associated with pancreas divisum (PD). A 60-year-old woman suffered from discomfort in the back of the right side. An abdominal radiograph revealed a calcified spherical mass in the right upper quadrant. Ultrasonography revealed a scattered echo with a posterior acoustic shadow in the gallbladder wall. A plain computed tomography (CT) scan showed flecks of intramural calcification in the wall of the gallbladder. Endoscopic retrograde cholangiopancreatography (ERCP) and magnetic resonance cholangiopancreatography (MRCP) showed separate openings for the Santorini and Wirsung ducts. The patient underwent cholecystectomy after porcelain gallbladder and pancreas divisum had been diagnosed. The porcelain gallbladder resulted from a stone impacted in the neck of the gallbladder. Patients with PD should be followed carefully, because gallstones often accompany PD, and porcelain gallbladder may result, as in this patient. PMID- 17139437 TI - Leiomyosarcoma of the superior mesenteric vein: a diagnostic and therapeutic challenge. AB - We report here a case of a very rare entity, a leiomyosarcoma of the mesentericoportal trunk, which was initially misdiagnosed as an unresectable pancreatic cancer invading the mesenteric vein, and which was finally treated by pancreatectomy with mesentericoportal reconstruction. The pitfalls of diagnosis and modalities of resection are discussed. PMID- 17139438 TI - Treatment of liver metastases from a solid pseudopapillary tumor of the pancreas. PMID- 17139439 TI - Gallbladder metastases from ductal papillary carcinoma of the breast. AB - Breast cancer occurs primarily in women aged 25 years or older. Breast carcinoma has the potential for widespread dissemination, such as metastasis to axillary lymph nodes, bone, lung, pleura, brain, and soft tissues. Liver, gastrointestinal, and biliary tract involvement are infrequent. We report a patient, a 62-year-old woman, with symptomatic cholelithiasis. The patient proceeded to laparoscopic cholecystectomy. She had a previous history of mastectomy with axillary lymphadenectomy, performed for a breast ductal papillary carcinoma, 10 years prior to the cholecystectomy. The gallbladder was hydropic; the wall was thickened, with a focal broad-based lesion on the mesenteric face of the body. Histopathological evaluation of the focal broad-based lesion of the gallbladder revealed poorly differentiated adenocarcinoma infiltration, without mucosal involvement. Subsequent immunohistochemical examination showed the lesion to be cytokeratin 7(CK7)-positive and cytokeratin 20 (CK20)-negative. Estrogen receptor (ER) and progesterone receptor (PgR) were positive. The final pathological diagnosis was breast ductal papillary carcinoma metastases to the gallbladder. Mammography of the other breast was normal. Computed tomography (CT) scan of the brain, chest, abdomen, and pelvis was performed, without any pathological findings. Bone Tc-99 scintigraphy was normal. Six months after the surgery positron emission tomography (PET) showed no evidence of metastatic disease. Two years after the surgery, the patient died, in the absence of recurrence. A literature review revealed only a few more cases of metastasic breast carcinoma to the gallbladder. PMID- 17139440 TI - Cystadenocarcinoma of the gallbladder. AB - Biliary cystadenocarcinomas are a very rare set of neoplasms that occur in the liver, the extrahepatic bile duct system, and occasionally in the gallbladder. We report on a 75-year-old man with a cystadenocarcinoma of the gallbladder who presented with mild abdominal discomfort and a palpable mass in the right upper abdomen. The diagnostic workup included ultrasound, magnetic resonance tomography, and fine needle core biopsy and revealed a multicystic tumor 16 cm in diameter. The patient underwent multivisceral resection of the tumor with a resection of the right colon, a partial resection of the stomach and the liver, and a lymph node dissection. The resected specimen weighed 2500 g and the final histopatholical diagnosis was a primary, moderately differentiated cystadenocarcinoma of the gallbladder with two lymph node metastases and invasion of the right colon. The patient remains without evidence of recurrence 12 months after surgery. The clinical presentation, diagnosis, therapeutic options, and classification of hepatobiliary cystadenocarcinomas are discussed. PMID- 17139441 TI - Disappearing cancer. PMID- 17139442 TI - Anaemia-related impairment in quality of life in elderly cancer patients prior to chemotherapy. AB - PURPOSE: Quality of life (QoL) of patients with cancer is a major area of concern for both patients and their physicians. PATIENTS AND METHODS: We investigated QoL with the EORTC-QLQ-C30 questionnaire with reference to anaemia and Karnofsky Performance Status (KPS) prior to the start of chemotherapy in 477 patients: 195 elderly cancer patients (Group A), 152 younger cancer patients (Group B), and 130 patients aged 60 years or older admitted for non-cancer related disorders (Group C). RESULTS: In univariate analysis QoL was significantly worse in 8 out of 15 scales in anaemic compared to non anaemic patients in Group A, in 2 in Group B, and in 7 in Group C. In ANOVA analysis including KPS and haemoglobin status, the influence of anaemia and KPS independently persists in most scales in Group A, in some in Group C, but not in Group B. CONCLUSIONS: Anaemia and functional impairment are independently related to QoL in elderly cancer and elderly medical patients, but not in younger cancer patients. PMID- 17139443 TI - Altered venous capacitance as a cause of postprandial hypotension in multiple system atrophy. AB - Patients with multiple system atrophy (MSA) often have clinically significant postprandial hypotension (PPH). To elucidate the cause of insufficient cardiac preload augmentation that underlies PPH, we recorded calf venous capacitance (CVC) by strain-gauge plethysmography, in 17 MSA patients and eight healthy controls before and after oral glucose ingestion. Among 17 MSA patients, nine who showed a decrease in systolic blood pressure exceeding 20 mmHg and were diagnosed with PPH. MSA patients without PPH showed a significant decrease in CVC and a significant increase in cardiac output after oral glucose ingestion, as did controls; those with MSA exhibiting PPH showed a significant increase in CVC and no significant change in cardiac output. The change in CVC correlated positively with the decrease in systolic and diastolic blood pressure after glucose ingestion, and also correlated negatively with the increase in cardiac output. Physiologically, PPH is prevented by a decrease in venous capacitance, which increases circulating blood volume and cardiac output. In some MSA patients, failure of venous capacitance to decrease may induce PPH. PMID- 17139445 TI - Molecular pharmacological approach to drug actions on the afferent and efferent fibres of the vagal nerve involved in gastric mucosal protection in rats. AB - BACKGROUND: Gastric mucosal protection is associated with the actions of anti ulcer drugs or agents affecting on the afferent and/or efferent nerve fibres of the vagal nerve. AIMS: 1. To identify the dose-response curves of drugs (compounds) on the afferent vanilloid-receptor (capsaicin or resiniferatoxin sensitive) and on efferent secretion (atropine, pirenzepine, cimetidine, ranitidine, famotidine, omeprazole, esomeprazole) basal gastric acid and stimulated gastric secretion in relation to the chemically-induced gastric mucosal damage in rats; 2. To determine the ED50 (pD2) and pA2 on the calculation of affinity and intrinsic affinity curves for these agonists/antagonists, as an indication of relative potency of effects. MATERIALS AND METHODS: The observations were carried out in rats (30 different models). RESULTS: The ED50 values for affinities of capsaicin, resiniferatoxin were obtained in nmol/kg b.w. range, whereas the values were in the nmol/kg to micromol/kg b.w. ranges for effects on the gastric basal, stimulated (bethanechol, pentagastrin, histamine) gastric secretion, and the gastric mucosal damage-produced by different ulcerogenic agents (ethanol, HCl, aspirin, indomethacin). CONCLUSION: From the observations, that agents acting on vanilloid (capsaicin) receptors were the most potent inhibitors of acid secretion and gastric lesions from necrotizing agents, suggests that the capsaicin sensitive afferent nerves have a primary place in the efferent regulated events leading to initiation of gastric mucosal damage. PMID- 17139446 TI - Rofecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor potentiates the anticonvulsant activity of tiagabine against pentylenetetrazol-induced convulsions in mice. AB - Numerous studies have implicated prostaglandins as potential modulators in seizure activity. The objective of the present study was to elucidate the effect of rofecoxib (selective COX-2 inhibitor) alone or in combination with newer antiepileptic drug tiagabine (gamma-amino acid reuptake inhibitor) against pentylenetetrazol (PTZ) (80 mg/kg, i.p.)-induced chemoconvulsions in mice. Rofecoxib or tiagabine was administered 45 min prior to the PTZ challenge. In combination study, rofecoxib was administered 10 min before tiagabine and after 35 min the animals were challenged with convulsive dose of PTZ. Mean onset time of jerks, clonus and extensor phases following PTZ challenge was noted. Pretreatment with rofecoxib (1-5.0 mg/kg, i.p.) or tiagabine (1-10 mg/kg, i.p.) dose dependently protected the animals against PTZ-induced convulsions. The mean onset time of jerks, clonus and extensor phase were increased. A subeffective dose of rofecoxib (0.5 mg/kg, i.p.) potentiated the effect of subprotective dose of tiagabine (0.5 mg/kg, i.p.). The results of the present study suggested that the protective effect of rofecoxib, a COX-2 inhibitor against PTZ-induced convulsions may be possibly through the GABAergic modulation. Rofecoxib may have a place as adjuvant therapy with standard antiepileptic drugs such as tiagabine in the treatment of epilepsy. PMID- 17139447 TI - Aspirin, TNF-alpha, NFkB, and survival in multiple myeloma: the importance of measuring TNF-alpha. AB - Aspirin and other cyclooxygenase inhibitors can increase levels of tumor necrosis factor-alpha and engage pro-apoptosis paths and/or anti-apoptosis paths. Seemingly conflicting data are briefly reviewed. Aspirin has been shown to slightly increase survival duration in multiple myeloma. In this brief note caution is raised about use of aspirin and COX inhibitors generally in inflammatory states and specifically in myeloma. Should they increase tumor necrosis factor-alpha they could exacerbate disease. A figure is presented showing the tendency for interleukin-6 and tumor necrosis factor-alpha, both demonstrated to be growth factors in myeloma, to be counter-regulated. Since both are now easily measured by specialty labs, it would be reasonable to monitor these during myeloma treatment generally, and particularly when using aspirin, COX inhibitors, or any other drug with potential to increase these growth factors. PMID- 17139448 TI - What is a gene? From molecules to metaphysics. AB - Mendelian genes have become molecular genes, with increasing puzzlement about locating them, due to increasing complexity in genomic webworks. Genome science finds modular and conserved units of inheritance, identified as homologous genes. Such genes are cybernetic, transmitting information over generations; this too requires multi-leveled analysis, from DNA transcription to development and reproduction of the whole organism. Genes are conserved; genes are also dynamic and creative in evolutionary speciation-most remarkably producing humans capable of wondering about what genes are. PMID- 17139449 TI - The question of questions: what is a gene? Comments on Rolston and Griffiths & Stotz. AB - If the question "What is a gene?'' proves to be worth asking it must be able to elicit an answer which both recognizes and address the reasons why the concept of the gene ever seemed to be something worth getting excited about in the first place as well analyzing and evaluating the latest develops in the molecular biology of DNA. Each of the preceding papers fails to do one of these and suffers the consequences. Where Rolston responds to the apparent failure of molecular biology to make good on the desideratum of the classical gene by veering off into fanciful talk about "cybernetic genes,'' Griffiths and Stotz lose themselves in the molecular fine print and forget to ask themselves why "genes'' should be of any special interest anyway. PMID- 17139450 TI - Glucosinolate hydrolysis in Lepidium sativum--identification of the thiocyanate forming protein. AB - Glucosinolates are a class of thioglycosides found predominantly in plants of the order Brassicales whose function in anti-herbivore defense has been attributed to the products formed by myrosinase-catalyzed hydrolysis upon plant tissue damage. The most common type of hydrolysis products, the isothiocyanates, are toxic to a wide range of organisms. Depending on the glucosinolate side-chain structure and the presence of certain protein factors, other types of hydrolysis products, such as simple nitriles, epithionitriles and organic thiocyanates, can be formed whose biological functions are not well understood. Of the proteins controlling glucosinolate hydrolysis, only epithiospecifier proteins (ESPs) that promote the formation of simple nitriles and epithionitriles have been identified on a molecular level. We investigated glucosinolate hydrolysis in Lepidium sativum and identified a thiocyanate-forming protein (TFP) that shares 63-68% amino acid sequence identity with known ESPs and up to 55% identity with myrosinase-binding proteins from Arabidopsis thaliana, but differs from ESPs in its biochemistry. TFP does not only catalyze thiocyanate and simple nitrile formation from benzylglucosinolate but also the formation of simple nitriles and epithionitriles from aliphatic glucosinolates. Analyses of glucosinolate hydrolysis products in L. sativum autolysates and TFP transcript accumulation revealed an organ-specific regulation of thiocyanate formation. The identification of TFP defines a new family of proteins that control glucosinolate hydrolysis and challenges the previously proposed reaction mechanism of epithionitrile formation. As a protein that promotes the formation of a wide variety of hydrolysis products, its identification provides an important tool for further elucidating the mechanisms of glucosinolate hydrolysis as well as the ecological role and the evolutionary origin of the glucosinolate-myrosinase system. PMID- 17139451 TI - The effect of Na+-K+ ATPase inhibition by ouabain on histamine release from human cutaneous mast cells. AB - BACKGROUND: There are controversial reports on the effect of sodium-potassium adenosine triphosphatase (Na+-K+ ATPase) inhibition on mast cell mediator release. Some of them have indicated that ouabain (strophanthin G), a specific Na+-K+ ATPase inhibitor, inhibited the release, whereas the others have shown that ouabain had no effect or even had a stimulatory effect on the mediator secretion. Most of these studies have utilized animal-derived mast cells. The aim of this study was to determine the effect of Na+-K+ ATPase inhibition on human skin mast cells. METHODS: Unpurified and purified mast cells were obtained from newborn foreskins and stimulated by calcium ionophore A23187 (1 microM) for 30 min following a 1 hr incubation with various concentrations (10(-4) to 10(-8) M) of ouabain. Histamine release was assayed by enzyme-linked immunosorbent assay (ELISA). RESULTS: The results indicated that ouabain had no significant effect on the non-immunologic histamine release from human skin mast cells, in vitro. CONCLUSIONS: Na+-K+ ATPase inhibition by ouabain had no significant effect on the non-immunologic histamine release from human cutaneous mast cells and suggested differences between human and animal mast cells. PMID- 17139452 TI - Isolation and characterization of the bovine microsatellite loci. AB - Microsatellite loci were isolated using five repetitive probes for Korean native cattle. Eleven microsatellite loci were developed based on a biotin hybrid capture method, and enrichment of the genomic libraries (AAAT, TG, AG, T, and TGC repeats) was performed using Sau3AI adapters. The isolated markers were tested in two half-sib Korean cattle families and four imported breeds (Angus, Limousine, Holstein, and Shorthorn). Nine informative microsatellite loci were observed, and two microsatellite loci were revealed as monomorphic in Korean cattle. In the imported breeds, however, all of the markers were informative. In total, 213 alleles were obtained at the 11 loci across five breeds, and the average number of alleles found per locus, considering all populations, was 4.26. Heterozygosity was 0.71 (expected) and 0.57 (observed). The range of the polymorphic information content for the markers in all cattle populations was 0.43-0.69. Eleven percent of genetic variation was attributed to differentiation between populations as determined by the mean F (ST) values. The remaining 89% corresponded to differences among individuals. The isolated markers may be used to identify and classify the local breeds on a molecular basis. PMID- 17139453 TI - The response of 2D foams to continuous applied shear in a Couette rheometer. AB - The continuum model that has reproduced the experimental observation of exponential shear localisation for straight-edge boundary conditions is adapted to the case of circular geometry. Essentially the same effect is found. However, the scenario of possible velocity profiles is much richer. Our calculations elucidate many recent experiments qualitatively and suggest further extensions of them. Various limits are analysed. In particular, the localisation length vanishes as the inner-boundary velocity tends to zero. PMID- 17139454 TI - Vorticity banding during the lamellar-to-onion transition in a lyotropic surfactant solution in shear flow. AB - We report on the rheology of a lyotropic lamellar surfactant solution (SDS/dodecane/pentanol/ water), and identify a discontinuous transition between two shear thinning regimes which correspond to the low-stress lamellar phase and the more viscous shear-induced multilamellar vesicle, or "onion" phase. We study in detail the flow curve, stress as a function of shear rate, during the transition region, and present evidence that the region consists of a shear banded phase where the material has macroscopically separated into bands of lamellae and onions stacked in the vorticity direction. We infer very slow and irregular transformations from lamellae to onions as the stress is increased through the two-phase region, and identify distinct events consistent with the nucleation of small fractions of onions that coexist with sheared lamellae. PMID- 17139455 TI - Laparoscopic resection with intraoperative radiotherapy: a new step in the multimodal treatment of advanced colorectal cancer. AB - BACKGROUND: Local recurrence is one of the most important problems related to resection of rectal cancer in locally advanced cases (T3-T4). Total mesorectal excision (TME) is the mainstay of surgical therapy, although many articles have been published about the availability of intraoperative radiotherapy (IORT) for the control of locally advanced rectal cancers. METHODS: The authors describe six patients affected by advanced rectal cancer (T3N1) whom they treated with neoadjuvant radiochemotherapy and laparoscopic rectal resection combined with TME and IORT. RESULTS: The operative time did not exceed 6 h in any case with IORT treatment. The procedure itself and the transfer of patients to the radiotherapy room accounted for about 2 h. The postoperative course was uneventful in every case, and all the patients were discharged within the first 8 postoperative days. CONCLUSIONS: This report describes the technical aspect and the feasibility of IORT associated with laparoscopic surgical resection for rectal cancer. PMID- 17139456 TI - The potential of restaging in the prediction of pathologic response after preoperative chemoradiotherapy for rectal cancer. AB - BACKGROUND: We performed this study to prospectively evaluate the postchemoradiotherapy performance of transrectal ultrasonography (TRUS), pelvic computed tomography (CT) scan and magnetic resonance imaging (MRI), and endoscopic biopsies for predicting the pathologic complete response of rectal cancer patients. METHODS: Four weeks after completion of preoperative chemoradiotherapy, 46 consecutive patients with mid to low rectal cancer were prospectively evaluated by proctoscopy, TRUS, and pelvic CT scan and MRI. On the basis of T and N status, patients were classified as T0 or T1-4 and N-negative or N-positive. For each staging modality used, sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were calculated. Findings were compared with the pathologic tumor-node-metastasis stage. RESULTS: On histopathologic analysis, 12 patients had pT0 and 34 had pT1-4 lesions; out of 45 assessable patients, 9 were N-positive. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy in predicting T status (T0 vs. T >or=1) were 77%, 33%, 74%, 36%, and 64%, respectively, for TRUS; 100%, 0%, 74%, not assessable, and 74% for CT; and 100%, 0%, 77%, not assessable, and 77% for MRI. The corresponding figures in predicting N status (N-negative vs. N positive) were, respectively, 37%, 67%, 21%, 81%, and 61% for TRUS; 78%, 58%, 32%, 91%, and 62% for CT; and 33%, 74%, 25%, 81%, and 65% for MRI. CONCLUSIONS: Current rectal cancer staging modalities after chemoradiotherapy allow good prediction of node-negative cases, although none of them is able to predict the pathologic complete response on the rectal wall. PMID- 17139457 TI - A population-based study on the management and outcome in patients with locally recurrent rectal cancer. AB - BACKGROUND: Although outcome in patients with rectal cancer has improved with preoperative radiotherapy and total mesorectal excision, local recurrence still remains a problem. The condition is difficult to cure and little is known on whether the prognosis for patients with locally recurrent tumours has changed over time. Few population-based studies have been performed. METHOD: Two thousand three hundred and eighteen patients in Stockholm, Sweden had a potentially curative resection for rectal cancer between 1995 and 2003. Until 2005, 141 (6%) developed a local recurrence. Management and outcome for these patients were studied and compared to a previously analysed cohort of 156 patients with local recurrence, treated 1980-1991. RESULTS: Of the 141 patients, 57 (40%) had surgery with a curative intent, 48 (34%) radio- and/or chemotherapy and 36 (26%) symptomatic palliation only. The total 5-year survival was 9%. Twenty-five patients had a potentially curative resection, with a 5-year survival of 57%. The corresponding figures for the 156 patients in the earlier cohort were 4 and 42%. CONCLUSION: Although outcome for patients with local recurrence of rectal cancer is dismal, the prognosis has improved slightly over time. A radical resection is a prerequisite for cure and the proportion having a potentially curative resection has increased. Multidisciplinary management, including optimised preoperative staging and patient selection for surgery, radical surgical approach and more effective adjuvant treatments are necessary to further improve the prognosis. PMID- 17139458 TI - Prognostic score in gastric cancer patients. PMID- 17139459 TI - Loco-regional recurrence from colon cancer: a population-based study. AB - BACKGROUND: The survival after colon cancer surgery has not improved to the same extent as after rectal cancer treatment and studies on loco-regional recurrence after colon cancer surgery are scarce. The aim of this study was to assess the problem of loco-regional recurrence after potentially curative resections for colon cancer, regarding incidence, risk factors, management, and outcome. METHODS: All 1,856 patients submitted to potentially curative surgery for colon cancer in the Stockholm/Gotland region in Sweden between 1996 and 2000 were followed until January 2005 or until death. Follow-up data were prospectively collected. Risk factors for loco-regional recurrences were analyzed, treatment and outcome for patients with recurrence was studied. RESULTS: The cumulative 5 year incidence of loco-regional recurrence was 11.5%. Tumor locations in the right flexure and in the sigmoid colon, bowel perforation and emergent surgery were identified as independent risk factors for loco-regional recurrence. The risk also increased with increasing T- and N-stage. The median survival for all 192 patients with loco-regional recurrence was 9 months. Surgery was performed in 110 (57%) patients. In 23 (12%) patients a complete tumor clearance was achieved and the estimated 5-year survival in this group was 43%. CONCLUSION: Loco regional recurrence from colon cancer is a significant clinical problem. A multidisciplinary treatment approach, including preoperative staging, a complete resection of the recurrence and more effective adjuvant treatments may improve the outcome. PMID- 17139460 TI - Incidence and treatment of tracheal cancer: a nationwide study in the Netherlands. AB - BACKGROUND: The aim of this study was to assess the incidence, characteristics, treatment, and survival of patients with tracheal malignancies in the Netherlands. METHODS: All cases of tracheal cancer entered into the database of the Netherlands Cancer Registry in the period 1989-2002 were selected. Data on histological type, age at time of diagnosis, treatment, and survival were analyzed retrospectively. RESULTS: The annual incidence was 0.142 per 100,000 inhabitants (308 cases, of which 15 were found incidentally at autopsy). Of these, 72% were men. In 52.9%, the histological type was squamous cell carcinoma and in only 7.1% adenoid cystic carcinoma (ACC). Mean age at time of diagnosis was 64.3 years. Of the 293 patients diagnosed while alive, 34 patients underwent surgical resection (11.6%), 156 patients received radiotherapy (53.2%), and 103 patients neither (35.4%). Median survival of all 293 patients was 10 months (mean 28 months) with 1-year, 5-year, and 10-year survival rates of 43%, 15%, and 6%, respectively. The prognosis of patients with ACC was significantly better. The 5 year survival rate in patients who underwent surgical resection was 51%, and the 10-year survival rate in these patients was 33%. CONCLUSION: The prognosis of patients with a tracheal malignancy is usually poor. Surgical treatment, however, can lead to good survival rates; still, this is currently only used in selected patients, even though it would seem to be possible in more cases in view of the technical advances in the field of tracheal surgery. Centralizing the care and treatment of tracheal cancers and implementing a more assertive attitude towards this disease could make surgery accessible to a larger number of patients. Data from the literature show that this would lead to better survival in patients with a tracheal malignancy. PMID- 17139461 TI - Surgical management after neoadjuvant imatinib therapy in gastrointestinal stromal tumours (GISTs) with respect to imatinib resistance caused by secondary KIT mutations. AB - BACKGROUND: In metastasized GISTs, resistance to imatinib after initial tumour response has been associated with observation of secondary mutations in the activation loop of KIT. The aim of the current study was to evaluate the tumour response and observance of secondary KIT mutations in a case of GIST undergoing neoadjuvant imatinib therapy. METHODS: We report on a case of an initially unresectable gastric GIST with curative resection after 10 months of neoadjuvant imatinib therapy. Mutation analysis of KIT was performed on a pretherapeutic biopsy specimen, as well as on the resected tumour specimen. RESULTS: The pretherapeutic biopsy revealed cKit positive tumour cells with mutation of KIT exon 11 Del 560-576. The remaining tumour mass after neoadjuvant imatinib therapy almost exclusively consisted of hypocellular myxohyalinale stroma with rare microfoci of cKit positive tumour cells. Laser microdissection of several tumour microfoci revealed two additional point mutations located in the activation loop of KIT exon 17, C809G and N822Y, each observed separately in a distinct microfocus. Neither of these two point mutations has been reported in a GIST so far. CONCLUSIONS: Neoadjuvant imatinib therapy successfully reduces tumour size in GISTs. Since resistance relevant secondary mutations of the activation loop of KIT may be observed after neoadjuvant imatinib therapy, the time elapse with preoperative imatinib therapy should be chosen as short as curative tumour resection or function sparing surgery can be carried out. The determination of the optimal time point for surgery is therefore a critical event and will be discussed. PMID- 17139462 TI - Is the role of Burch colposuspension fading away in this epoch for treating female urinary incontinence? AB - The role of Burch colposuspension as the primary surgical treatment of stress urinary incontinence has been challenged by less invasive new surgical methods. The aim of this study was to evaluate the long-term results of Burch colposuspension in terms of subjective self-reported outcomes. Between 1993 and 1997, 159 women who underwent Burch colposuspension as the first operation for treating urodynamic stress incontinence were recruited for this study. We recorded the findings of preoperative and postoperative urodynamic studies and early postoperative complications or adverse effects related to the operation. In 2005, after a median follow-up of 10 years, telephone interviews were carried out and 152 (95.5%) women responded. Two main questions were asked of these women to evaluate the overall impression of improvement after the operation. Eighty-four (55.3%) women were dry according to their subjective reports, 55 (36.2%) women had improved, and 13 (8.5%) women had failed after an 8- to 12-year follow-up. One hundred and twenty-five (82.2%) women were satisfied with the outcome of the operation and 27 (17.8%) women were not. Among these 27 women, 16 (59.2%) women complained of urinary frequency and 9 (33.3%) women complained of urinary urgency as the reasons for their dissatisfaction. Our long-term subjective outcomes revealed that Burch colposuspension is an effective alternative surgery for urodynamic proven stress incontinence. PMID- 17139463 TI - Postural activity of the pelvic floor muscles is delayed during rapid arm movements in women with stress urinary incontinence. AB - The aim of this study was to determine whether postural activity of the pelvic floor (PF) and abdominal muscles differs between continent and incontinent women during rapid arm movements that present a postural challenge to the trunk. A further aim was to study the effect of bladder filling. Electromyographic activity (EMG) of the PF, abdominal, erector spinae (ES), and deltoid muscles was recorded with surface electrodes. During rapid shoulder flexion and extension, PF EMG increased before that of the deltoid in continent women, but after the deltoid in incontinent women (p = 0.002). In many incontinent women, PF EMG decreased before the postural activation. Although delayed, postural PF EMG amplitude was greater in women with incontinence (p = 0.010). In both groups, PF EMG decreased and abdominal and ES EMG increased when the bladder was moderately full. These findings would be expected to have negative consequences for continence and lumbopelvic stability in women with incontinence. PMID- 17139464 TI - MRI-measured bone marrow adipose tissue is inversely related to DXA-measured bone mineral in Caucasian women. AB - INTRODUCTION: Recent studies suggest that bone marrow adipose tissue (BMAT) might play a role in the pathogenesis of osteoporosis. Previous research using regional magnetic resonance spectroscopy methods to measure BMAT has reported inconsistent findings on the relationship between BMAT and dual-energy absorptiometry (DXA) measured bone mineral density (BMD). METHODS: In the present study, total body and pelvic BMAT were evaluated in 56 healthy women (age 18-88 yrs, mean +/- SD, 47.4 +/- 17.6 yrs; BMI, 24.3 +/- 4.2 kg/m(2)) with T1-weighted whole-body magnetic resonance imaging (MRI). BMD was measured using the whole-body DXA mode (GE Lunar DPX, software version 4.7). RESULTS: A strong negative correlation was observed between pelvic BMAT and BMD (total-body BMD, R = -0.743, P < 0.001; pelvic BMD, R = -0.646, P < 0.001), and between total-body BMAT and BMD (total body BMD, R = -0.443, P < 0.001; pelvic BMD, R = -0.308, P < 0.001). The inverse association between pelvic BMAT and BMD remained strong after adjusting for age, weight, total body fat, and menopausal status (partial correlation: total-body BMD, R = -0.553, P < 0.001; pelvic BMD, R = -0.513, P < 0.001). BMAT was also highly correlated with age (pelvic BMAT, R = 0.715, P < 0.001; total-body BMAT, R = 0.519, P < 0.001). CONCLUSION: MRI-measured BMAT is thus strongly inversely correlated with DXA-measured BMD independent of other predictor variables. These observations, in the context of DXA technical concerns, support the growing evidence linking BMAT with low bone density. PMID- 17139466 TI - Evaluation of clinical prognosis and activities of daily living using functional independence measure in patients with hip fractures. AB - BACKGROUND: The functional independence measure (FIM) is an evaluation method of activities of daily living (ADL) that assesses motor functions and cognitive functions in the Uniform Data System. The FIM has recently been used to assess disability. The purpose of this study was to standardize criteria using the FIM for determining when and to where patients can be discharged following surgery for hip fracture. METHODS: Patients with hip fracture (n=68) aged>or=65 years who underwent surgery at our hospital were classified by their residence at the time of injury (their own home, a hospital, or an elderly care facility) and by postoperative residence after discharge from hospital. We investigated the FIM of these patients before injury and at the time of discharge and retrospectively compared the results with the Japan Orthopaedic Association (JOA) hip score at the time of discharge. RESULTS: Patients who entered a facility after discharge following surgery demonstrated a reduction in motor function score on the FIM. Cognitive function scores in each group were not reduced postoperatively in the short term. The average reduction in scores on the FIM for patients who were discharged from hospital to their own home was 15.9 points, and it was 25.9 points for those who were injured in their own home and transferred to a facility after discharge. There was a significant correlation between the FIM and the JOA hip score at the time of discharge. CONCLUSIONS: The FIM cannot determine whether such patients should be discharged to their home or transferred to a care facility. However, the motor function scores on the FIM are valid for assessing hip fracture patients and may be suitable as a standardized procedure for determining their postdischarge residence. PMID- 17139467 TI - Single nucleotide polymorphisms in the interleukin-6 gene promoter, tumor necrosis factor-alpha gene promoter, and transforming growth factor-beta1 gene signal sequence as predictors of time to onset of aseptic loosening after total hip arthroplasty: preliminary study. AB - BACKGROUND: Aseptic loosening resulting from inflammatory response to the implant wear debris is the major cause of late total hip arthroplasty (THA) failure. We examined single nucleotide polymorphisms in genes encoding for three involved cytokines--interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and transforming growth factor-beta1 (TGF-beta1)--as potential predictors of time to onset of aseptic instability. METHODS: A total of 41 patients/45 total hip endoprostheses (same type, same surgeon) were followed up for as long as 18 years. They were genotyped for the IL-6 promoter (-597G-->A) and (-572G-->C), TNF alpha promoter (-308G-->A), and TGF-beta1 signal sequence (29T-->C) transitions. Cox regression was performed on the prosthesis survival. RESULTS: Overall, 22 of 45 prostheses developed aseptic instability. Cumulative survivals at 10 and 15 years after THA were 95.6% and 66.6%, respectively. The effect of a particular polymorphic site was estimated with adjustment for sex, age at THA, reason for THA, and the effects of other analyzed sites. The hazard ratio (HR) for genotype T/T versus "C-allele carriage" at the TGF-beta1 site was 8.23 [95% confidence interval (CI) 1.45-46.8] (P=0.017) or 5.70 (1.39-23.4) (P=0.016) when the IL-6 promoter sites were considered as a "combination of genotypes (-597)|(-572)." The most prevalent combination of genotypes at IL-6 sites was G/A (-597)|C/C (-572). HR for this combination (versus other combinations) was 5.43 (1.73-17.0) (P=0.004) when "TGF-beta1 (29T-->C)" was considered as a three-level factor (three possible genotypes), and 4.92 (1.71-14.1) (P=0.003) when TGF-beta1 site was considered as a two-level factor (T/T and "C-allele carriage"). The HR for the "A-allele carriage" at TNF-alpha (-308G-->A) could not be determined (only two patients had the G/G genotype). CONCLUSIONS: This preliminary study is the first to suggest that the TGF-beta1 signal sequence (29T-->C) and IL-6 promoter ( 597G-->A)|(-572G-->C) transitions are predictive for the time to onset of aseptic instability after THA. PMID- 17139468 TI - Three-dimensional knee motion before and after high tibial osteotomy for medial knee osteoarthritis. AB - BACKGROUND: High tibial osteotomy (HTO) is an established surgical option for treating medial knee osteoarthritis. HTO moves the mechanical load on the knee joint from the medial compartment to the lateral compartment by changing the leg alignment, but the effects of the operation remain unclear. The purpose of this study was to evaluate the change in three-dimensional knee motion before and after HTO, focusing on lateral thrust and screw home movement, and to investigate the relationship between the change in knee motion and the clinical results. METHODS: A series of 19 patients with medial knee osteoarthritis who had undergone HTO were evaluated. We performed a clinical assessment, radiological evaluation, and motion analysis at 2.4 years postoperatively. The clinical assessment was performed using the Japanese Orthopaedic Association knee score. RESULTS: The score was significantly improved in all patients after operation. Motion analysis revealed that lateral thrust, which was observed in 18 of the 20 knees before operation, was reduced to 7 knees after operation. Regarding active terminal extension of the knee, three patterns of rotational movement were observed before operation: screw home movement (external rotation), reverse screw home movement (internal rotation), and no rotation. By contrast, after operation, only reverse screw home movement and no rotation were observed; the screw home movement disappeared in all patients. In the knees with reverse screw home movement after operation, the preoperative score was significantly lower than those in the knees with no rotation after operation. CONCLUSIONS: Kinetically, HTO was useful for suppressing lateral thrust in medial knee osteoarthritis, although the rotational movement of the knee joint was unchanged. PMID- 17139469 TI - Histologic classification of loose bodies in osteoarthrosis. AB - BACKGROUND: Histologically based analyses of the nature and origin of loose bodies occurring in osteoarthrosis have been few, and further study is warranted. METHODS: We histologically examined 84 loose bodies and 9 related lesions (synovial membrane nodules) surgically removed from 24 joints of 24 patients with osteoarthrosis. RESULTS: The 84 loose bodies included 48 chondral loose bodies (type I), 26 osteochondral loose bodies (type II), and 10 osseous loose bodies (type III). The 26 osteochondral loose bodies (type II) could be subdivided into 8 composed of cartilage with enchondral ossification (type IIa), 11 consisting of mature bone covered by cartilage without enchondral ossification (type IIb), and 7 made up of mature bone and partially articular cartilage or hyaline cartilage (type IIc). Synovial membrane nodules could be also divided into three types in the same manner as loose bodies. Many type IIa, type IIc and type III loose bodies and all synovial membrane nodules showed blood vessels containing red blood cells, as well as osteoclasts. The type I and type IIb loose bodies, however, did not show them. CONCLUSIONS: It is well known that loose bodies grow from proliferation of cartilage without blood supply in the joint cavity, and that enchondral ossification is able to develop only under the condition of having a blood supply. As synovial membrane nodules were also classified to the same types as loose bodies and more than half of osteochondral and osseous loose bodies contained blood vessels with red blood cells, the loose bodies were thought to be caught in the synovial membrane and to be modified as the result of a blood supply. Considering the results of this study, the various histologic characteristics of loose bodies in osteoarthrosis resulted from modifications including cartilage proliferation in the joint cavity and enchondral ossification in the synovial membrane. PMID- 17139470 TI - Biomechanical differences resulting from the combination of suture materials and repair techniques. AB - BACKGROUND: Many suture materials and repair techniques have been applied in clinical tendon surgery. However, no recommendation is available concerning the choice of suture materials and repair techniques except in a few experimental studies. The purpose of the current study is to show the biomechanical difference resulting from the combination of suture materials and repair techniques. METHODS: The gastrocnemius tendons of 24-week-old cattle (diameter 14-16x9-11 mm) were repaired with application of a single locking, multiple locking, single grasping, or multiple grasping technique using a USP2 suture thread of either braided polyblend polyethylene, polyester, polydioxanone, or nylon. Therefore, a total of 16 combinations were made, with eight specimens for each combination. The specimen was set in an Instron tensiometer to measure the gap length after repetitive tensile loading 500 times (10-100 N). RESULTS: The single locking technique using braided polyblend polyethylene provided the smallest gap (4.5+/ 0.5 mm). Other techniques using the same material resulted in a large gap (10.0 11.8 mm). The polyester provided a relatively smaller gap length, irrespective of the repair technique (7.4-8.8 mm). Polydioxanone and nylon tended to result in a large gap (9.3-12.3 mm and 8.4-10.6 mm, respectively). CONCLUSIONS: Mechanical properties of each tendon suture depended on the particular combination of suture materials and repair techniques. The combination of braided polyblend polyethylene and single locking technique provided the highest antigap strength. PMID- 17139471 TI - Dose-related effects of recombinant human interleukin-10 on hypoxia-induced skeletal muscle injury in immature rats. AB - BACKGROUND: The role of cytokines in the pathogenesis of skeletal muscle injury in patients with hypoxia and inflammation remains unclear. The aim of the present study was to determine the dose-related effects of recombinant human interleukin 10 (rhIL-10) on hypoxia-induced skeletal muscle injury in immature rats. METHODS: The study was performed on 1-day-old Sprague-Dawley rat pups that were randomized to one of five groups: nonhypoxic controls (group 1; n=8); rats subjected to hypoxia-reoxygenation (H/O) and returned to their mothers (group 2; n=11); rats subjected to H/O, returned to their mothers, and treated with rhIL-10 (10 microg/kg per day s.c.) for 3 days (group 3; n=10); rats subjected to H/O, returned to their mothers, and treated with rhIL-10 (25 microg/kg per day s.c.) for 3 days (group 4; n=10); or rats subjected to H/O, returned to their mothers, and treated with rhIL-10 (75 microg/kg per day s.c.) for 3 days (group 5; n=10). All animals were killed on day 4, and skeletal muscle specimens were obtained to determine the tissue levels of malondialdehyde (MDA) and superoxide dismutase (SOD) and to check for any histological changes. RESULTS: The polymorphonuclear leukocyte count, amount of interstitial edema, microvessel size, and area of myocyte necrosis was greater in groups 2, 3, 4, and 5 than in group 1, although the findings were significantly less prominent in groups 4 and 5 than in groups 2 and 3. MDA levels significantly increased in groups 2 and 3 compared to those in groups 1, 4, and 5. SOD values decreased significantly in groups 2 and 3 compared to those in groups 1, 4, and 5. CONCLUSIONS: Administration of rhIL-10 reduced skeletal muscle injury produced by hypoxia in immature rats. Intermediate and high doses of rhIL-10 were associated with a significant reduction of skeletal muscle damage, whereas a low dose was not. PMID- 17139472 TI - Safe, rapid, and effortless femoral nail removal using a new third-generation universal femoral nail extraction tool. PMID- 17139473 TI - Absence of osteonecrosis of the femoral head following bilateral femoral neck fracture with a high degree of displacement. PMID- 17139474 TI - Histopathological study of osteonecrosis 19 years after transtrochanteric rotational osteotomy. PMID- 17139475 TI - Implant failure of long Gamma nail in a patient with intertrochanteric subtrochanteric fracture. PMID- 17139476 TI - Destructive spondyloarthropathy of the atlantoaxial joint with severe spinal cord compression in long-term hemodialysis patients. PMID- 17139477 TI - Treatment of nonunion of the femoral neck fracture associated with avascular necrosis in a child. PMID- 17139478 TI - Pathological findings of an autograft containing osteosarcoma treated by liquid nitrogen retrieved 2 years after implantation. PMID- 17139480 TI - Cardiac metabolism and mechanics are altered by genetic loss of lipoprotein triglyceride lipolysis. AB - INTRODUCTION: Most circulating fatty acids are contained in lipoprotein triglycerides. For the heart to acquire these lipids, they must be broken down into free fatty acids via the enzyme lipoprotein lipase (LpL). Although it has long been known that hearts primarily use esterified fatty acids as fuel, different sources of fatty acids were thought to be interchangeable. MATERIALS AND METHODS: By creating mice with neonatal and acute LpL deletion we showed that lipoprotein-derived fatty acids could not be replaced by albumin-associated free fatty acids. Loss of cardiac LpL forces the heart to increase its uptake of glucose, reduce fatty acid oxidation, and eventually leads to cardiac dysfunction. In contrast, cardiomyocyte specific overexpression of an anchored form of LpL leads to excess lipid uptake, induction of fatty acid oxidation genes, and dilated cardiomyopathy. CONCLUSION: Increasing lipid secretion from the heart or redirecting lipids to adipose tissue can alleviate this lipotoxic situation. PMID- 17139479 TI - Intramedullary nailing of femoral and tibial shaft fractures. PMID- 17139481 TI - Effect of HPr phosphorylation on structure, dynamics, and interactions in the course of transcriptional control. AB - The serine46-phosphorylated form of the bacterial protein HPr fulfils an essential function in carbon catabolite repression (CCR). Using molecular dynamics (MD) we studied the effect of Ser46 phosphorylation on the molecular properties of HPr and its capability to act as the co-repressor of carbon catabolite protein A (CcpA). The calculated pK (a) values for a representative set of HPr(Ser46P) structures indicate that the phosphate group of HPr(Ser46P) exists predominantly in the unprotonated form under neutral conditions. A hydrogen bond detected in HPr(Ser46P) between one phosphate-group oxygen and a side-chain hydrogen of Asn43-an amino acid conserved in all HPr proteins of Gram positive bacteria that regulate their carbon consumption by CCR-might fulfil an important role in CcpA-HPr(Ser46P) complex formation. MD simulations show that the Ser46P-Asn43 hydrogen bond present in the unbound structure is replaced by intermolecular interactions upon complex formation. The degree to which amino acids in the CcpA-HPr(Ser46P) interface contribute to cofactor binding was analyzed by in silico alanine scanning. Lys307, Arg303, Asp296, Val300, and Tyr295 of CcpA were identified as important amino acids for the CcpA-HPr(Ser46P) interaction. Three of these residues are directly involved in sensing the correct phosphorylation state at His15(HPr) and Ser46(HPr). A substitution of interface residues Val319, Val314, Ser316, Leu321 and Gln320 by alanine showed that these amino acids, which contact helix alpha2 of HPr(Ser46P), play a less prominent role for complex formation. PMID- 17139482 TI - Quorum sensing: bacterial chatting. PMID- 17139483 TI - Analysis of the hierarchy of quorum-sensing regulation in Pseudomonas aeruginosa. AB - Quorum-sensing in Pseudomonas aeruginosa is known to regulate several aspects of pathogenesis, including virulence factor production, biofilm development, and antimicrobial resistance. Recent high-throughput analysis has revealed the existence of several layers of regulation within the QS-circuit. To address this complexity, mutations in genes encoding known or putative transcriptional regulators that were also identified as being regulated by the las and/or rhl QS systems were screened for their contribution in mediating several phenotypes, for example motility, secreted virulence products, and pathogenic capacity in a lettuce leaf model. These studies have further elucidated the potential contribution to virulence of these genes within the QS regulon. PMID- 17139484 TI - Electron-impact and glow-discharge ionization LC-MS analysis of green tea tincture. AB - A liquid chromatography-particle-beam mass spectrometer (LC-PB/MS) with interchangeable electron-impact (EI) and glow-discharge (GD) ion sources was evaluated for future application in analysis of botanical extracts. In this work a green tea tincture was characterized for a series of catechin components (catechin, epicatechin, epigallocatechin, and epigallocatechin gallate (EGCG)) and caffeine. Special emphasis was given to EGCG and caffeine, because they are important in determining the possible health effects of the green tea. The effects of instrument operating conditions were evaluated for the EI and GD ionization sources to determine their effect on analyte intensities and fragmentation patterns. These studies furnished information about the effects of these conditions in determining possible ionization pathways in the two ion sources. The mass spectra of these compounds obtained with the GD ion source are EI-like in appearance, with clearly identified molecular ions and fragmentation patterns that are easily rationalized. The absolute limits of detection for EGCG and caffeine were, respectively, 11 ng and 0.77 ng for the EI source and 3.2 ng and 0.61 ng for the GD source. The PB/EIMS and PB/GDMS combinations can be operated in a flow-injection mode, wherein the analyte is injected directly into the mobile phase, or coupled to high-performance liquid chromatography (HPLC), enabling LC-MS analysis of complex mixtures. A reversed-phase chromatographic separation of the green tea tincture was performed on a commercial C18 column using a gradient of water (containing 0.1% TFA) and ACN. Quantification of EGCG and caffeine was performed by the standard addition method. The amounts of EGCG and caffeine in the tested green tea tincture were each approximately 14 mg mL-1. PMID- 17139485 TI - Sensor technology and its application in environmental analysis. AB - Environmental analysis is one of the fundamental applications of chemical sensors. In this review we describe different sensor systems for the gas and liquid phases that have been tested either with real-life samples or in the field during the last five years. Most field sensors rely either on electrochemical or optical transducers. In the gas phase, systems have been proposed for analysis of oxides of nitrogen, carbon, and sulfur in air, and volatile organic compounds. In the liquid phase, most detection systems used for real-life samples detect heavy metal ions or organic contamination, for example pesticides, organic solvents and polycyclic aromatic hydrocarbons. PMID- 17139486 TI - Characterisation of betalain patterns of differently coloured inflorescences from Gomphrena globosa L. and Bougainvillea sp. by HPLC-DAD-ESI-MSn. AB - In the present study, the betaxanthin (bx) and betacyanin patterns of differently coloured inflorescences from Gomphrena globosa L. and Bougainvillea sp. have been investigated in detail by applying reversed phase high-performance liquid chromatography-diode array detection (HPLC-DAD) coupled with positive ion electrospray mass spectrometry. Histidine-bx was found to be the predominant betaxanthin of Gomphrena globosa inflorescences. Furthermore, arginine-bx was detected as a novel betaxanthin, which to the best of our knowledge has not been reported as a pigment that occurs naturally so far. Dopa-bx was the major betaxanthin of Bougainvillea sp., although several minor betaxanthins were also present, including lysine-bx and putrescine-bx, novel betaxanthins hitherto not observed naturally. Remarkable differences in the betacyanin patterns between the purple, red and orange varieties were observed for both Gomphrena and Bougainvillea inflorescences. Hence, both the betacyanin profiles and the relative betaxanthin:betacyanin ratios determine the broad colour palette of Gomphrena petals and Bougainvillea bracts. PMID- 17139487 TI - Sensitive measurement of ultratrace phenols in natural water by purge-and-trap with in situ acetylation coupled with gas chromatography-mass spectrometry. AB - A novel purge-and-trap method coupled with gas chromatography-mass spectrometry (GC-MS) is developed for the analysis of trace and ultratrace phenols based on their derivatization with acetic anhydride. Parameters affecting the extraction efficiency, such as purge temperature, concentration of sodium chloride, purge time, and volume of derivatization reagent, were investigated. The optimized conditions were addition of 150 microL acetic anhydride, purge time of 25 min at the purge temperature of 60 degrees C with 30% NaCl. The linear range was 0.2-100 microg L(-1) for phenols. The limits of detection (LODs) ranged from 0.08 to 0.15 microg L(-1) and the relative standard deviations (RSDs) for most of the phenols at the 10 microg L(-1) level were below 10%. Natural water samples collected from a pool were successfully analyzed using the proposed method. The recovery of spiked water samples was 72.9-84.2%. PMID- 17139488 TI - Determination of non-steroidal anti-inflammatory drugs in environmental samples by chromatographic and electrophoretic techniques. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are a group of pharmaceutical compounds widely used in human health care and often found in the aquatic environment, with their metabolites. After an introduction that describes the general problem of drug contamination, the properties of NSAIDs, and environmental risk assessment, this review surveys the chromatographic and electrophoretic methods of analysis in use today for monitoring the most important representatives of this pharmaceutical class in different environmental samples. PMID- 17139489 TI - Unilateral primary ovarian leiomyoma in adolescent: a case report. AB - BACKGROUND: Primary ovarian leiomyoma is a rare, usually small and incidentally detected neoplasm, clinically indistinguishable from subserous leiomyomas and ovarian fibromas, until histopathological confirmation. CASE REPORT: We report a rare case of unilateral, ovarian leiomyoma that was not associated with uterine tumor in a 17-year-old woman. Trans abdominal ultrasonography and magnetic resonance imaging (MRI) revealed a pelvic mass (15 cm x 12 cm). CONCLUSION: At laparotomy, unilateral salpingo-oophorectomy was performed and histologic examination revealed a leiomyoma arising primarily in the ovary. PMID- 17139490 TI - Intravitreal bevacizumab for exudative age-related macular degeneration after multiple treatments. AB - BACKGROUND: Photodynamic therapy with verteporfin (PDT) significantly reduces the risk of vision loss in patients with exudative age-related macular degeneration (AMD). Indocyanine green-mediated photothrombosis (IMP) and trans-pupillary thermotherapy (TTT) may also be beneficial for selective cases of exudative AMD. However, a substantial subset of patients responds poorly to these treatments. Intravitreal bevacizumab (IVB) has been recently used in the treatment of exudative AMD, showing both visual and anatomic improvement in the majority of cases. METHODS: This interventional retrospective case series reports the effects of IVB in 17 eyes with subfoveal neovascular AMD that had undergone repeated PDT (combined or not with triamcinolone acetonide) or PDT followed by either IMP or TTT with poor results. The main outcome measures were visual acuity and tomographic signs of intra/subretinal fluid, as well as central retinal thickness. RESULTS: Most patients received a single IVB injection. The mean follow-up was 4.47 months. The mean logMAR visual acuity changed from 1.17 +/- 0.40 to 1.06 +/- 0.44 (P = 0.17). The mean central retinal thickness decreased from 404.05 +/- 245.26 to 280.23 +/- 143.14 microm (P = 0.032). At the end of the study, lack of tomographic signs of intra/ subretinal fluid was noted in four (23.5%) eyes. No ocular or systemic side effects were identified. CONCLUSIONS: Short-term results with IVB for the treatment of exudative AMD have been promising. However, the chronic retinal and pigment epithelium changes frequently present in eyes that underwent multiple previous treatments may limit complete visual recovery. To our knowledge, this is the first report on the use of IVB for this particular group of AMD patients. PMID- 17139492 TI - TLR7 imidazoquinoline ligand 3M-019 is a potent adjuvant for pure protein prototype vaccines. AB - Cancer vaccines, while theoretically attractive, present difficult challenges that must be overcome to be effective. Cancer vaccines are often poorly immunogenic and may require augmentation of immunogenicity through the use of adjuvants and/or immune response modifiers. Toll-like receptor (TLR) ligands are a relatively new class of immune response modifiers that may have great potential in inducing and augmenting both cellular and humoral immunity to vaccines. TLR7 ligands produce strong cellular responses and specific IgG2a and IgG2b antibody responses to protein immunogens. This study shows that a new TLR7 ligand, 3M-019, in combination with liposomes produces very strong immune responses to a pure protein prototype vaccine in mice. Female C57BL/6 mice were immunized subcutaneously with ovalbumin (OVA, 0.1 mg/dose) weekly 4x. Some groups were immunized to OVA plus 3M-019 or to OVA plus 3M-019 encapsulated in liposomes. Both antibody and cellular immune responses against OVA were measured after either two or four immunizations. Anti-OVA IgG antibody responses were significantly increased after two immunizations and were substantially higher after four immunizations in mice immunized with OVA combined with 3M-019. Encapsulation in liposomes further augmented antibody responses. IgM responses, on the other hand, were lowered by 3M-019. OVA-specific IgG2a levels were increased 625-fold by 3M-019 in liposomes compared to OVA alone, while anti-OVA IgG2b levels were over 3,000 times higher. In both cases encapsulation of 3M-019 in liposomes was stronger than either liposomes alone or 3M-019 without liposomes. Cellular immune responses were likewise increased by 3M-019 but further enhanced when it was encapsulated in liposomes. The lack of toxicity also indicates that this combination may by safe, effective method to boost immune response to cancer vaccines. PMID- 17139493 TI - Small numbers of residual tumor cells at the site of primary inoculation are critical for anti-tumor immunity following challenge at a secondary location. AB - Luciferase-transduced B16 murine melanoma cells (luc-B16) inoculated in ear skin do not form tumors but prevent tumor formation by luc-B16 cells injected into the footpad. To determine the requirements for such immunity, we followed the fate of luc-B16 cells following ear injection. Surprisingly, small numbers of viable luc B16 cells were detected in tumor-free mouse skin for up to 60 days post inoculation. After 1 week, the number of Foxp3(+)CD4(+)CD25(+) T cells (along with foxp3 mRNA expression) increased rapidly in the injected ear skin. Residual tumor cells in ears were reduced in mice treated with anti-CD25 mAb and in CD4 deficient mice, but increased in CD8-deficient mice. Strikingly, the loss of luc B16 cells in the ear skin, either spontaneously or following amputation of the injected ear, resulted in significantly enhanced tumor formation by parental and luciferase-expressing B16 cells after footpad injection. These studies suggest that small numbers of tumor cells (possibly regulated by CD4(+)CD25(+) regulatory T cells expressing Foxp3) are required for effective host anti-tumor responses at alternate inoculation sites. PMID- 17139494 TI - The emerging role of cannabinoid neuromodulators in symptom management. AB - INTRODUCTION: The cannabinoids nabilone (Cesamet) and dronabinol (Marinol) are indicated for the management of chemotherapy-induced nausea and vomiting (CINV) in cancer patients who have failed to respond adequately to conventional antiemetic therapy. DISCUSSION: The endocannabinoid (CB) system interacts with numerous other systems and pharmaceutical cannabinoids target ubiquitous CB1 and CB2 receptors in the central nervous system and periphery, relieving nausea and vomiting and pain. SUMMARY: The benefits of this novel class of medications in cancer may extend beyond CINV, as indicated by data from preclinical studies and animal models. PMID- 17139495 TI - Oral mucositis and outcomes of allogeneic hematopoietic stem-cell transplantation in patients with hematologic malignancies. AB - GOALS OF THE WORK: To assess the relationship between oral mucositis (OM) and adverse clinical and economic outcomes in patients with hematologic malignancies receiving allogeneic hematopoietic stem-cell transplantation (HSCT). MATERIALS AND METHODS: A retrospective chart review study of 281 allogeneic HSCT recipients with hematologic malignancies was undertaken at a single academic center. OM extent and severity were assessed across eight oropharyngeal sites using a validated scale, which was scored as follows: no erythema/ulceration=0; erythema only=I; ulceration, one site=II; ulceration, two sites=III; ulceration, three sites=IV and ulceration, four or more sites=V. OM assessments began on the day of conditioning and continued twice weekly within 28 days or hospital discharge. Analyses examined the relationship between the worst OM grade and selected adverse outcomes, including days with fever, days of total parenteral nutrition (TPN), days of parenteral narcotic therapy, incidence of significant (common terminology criteria (CTC) grade 3 or 4) infection, mortality and inpatient days and charges. MAIN RESULTS: The mean age of the study subjects was 41 years. Of the patients, 96% (n = 269) received total body irradiation and 76% (n = 214) experienced an OM grade of > or =II (i.e., ulceration). The worst OM grade was significantly (p < 0.05) associated with the number of days of TPN and parenteral narcotic therapy, number of days with fever, incidence of significant infection, time in hospital and total inpatient charges. CONCLUSIONS: OM is associated with worse clinical and economic outcomes in patients with hematologic malignancies undergoing allogeneic HSCT. PMID- 17139496 TI - The pharmacological importance of cytochrome CYP3A4 in the palliation of symptoms: review and recommendations for avoiding adverse drug interactions. AB - BACKGROUND: Adverse drug interactions are major causes of morbidity, hospitalizations, and mortality. The greatest risk of drug interactions occurs through in the cytochrome system. CYP3A4, the most prevalent cytochrome, accounts for 30-50% of drugs metabolized through type I enzymes. MATERIALS AND METHODS: Palliative patients received medications for symptoms and co-morbidities, many of which are substrate, inhibitors, or promoters of CYP3A4 activity and expression. A literature review on CYP3A4 was performed pertinent to palliative medicine. DISCUSSION: In this state of the art review, we discuss the CYP3A4 genetics, and kinetics and common medications, which are substrates or inhibitor/promoters of CYP3A4. CONCLUSION: We made some recommendations for drug choices to avoid clinically important drug interaction. PMID- 17139498 TI - Biomolecules preserved in ca. 168 million year old fossil conifer wood. AB - Biomarkers are widely known to occur in the fossil record, but the unaltered biomolecules are rarely reported from sediments older than Paleogene. Polar terpenoids, the natural products most resistant to degradation processes, were reported mainly from the Tertiary conifers, and the oldest known are Cretaceous in age. In this paper, we report the occurrence of relatively high concentrations of ferruginol derivatives and other polar diterpenoids, as well as their diagenetic products, in a conifer wood Protopodocarpoxylon from the Middle Jurassic of Poland. Thus, the natural product terpenoids reported in this paper are definitely the oldest polar biomolecules detected in geological samples. The extracted phenolic abietanes like ferruginol and its derivatives (6,7 dehydroferruginol, sugiol, 11,14-dioxopisiferic acid) are produced only by distinct conifer families (Cupressaceae s. l., Podocarpaceae and Araucariaceae), to which Protopodocarpoxylon could belong based on anatomical characteristics. Therefore, the natural product terpenoids are of great advantage in systematics of fossil plant remains older than Paleogene and lacking suitable anatomical preservation. PMID- 17139499 TI - Life on the edge: African malaria mosquito (Anopheles gambiae s. l.) larvae are amphibious. AB - Anopheles gambiae s.l. is the main vector of malaria in Sub-Saharan Africa. Here, an estimated 1 million people die every year from this disease. Despite considerable research on An. gambiae that increasingly explores sub-organismal phenomena, important facets of the field biology of this deadly insect are yet being discovered. In the current study, we used simple observational tools to reveal that the habitat of larval An. gambiae is not limited within the boundaries of temporary mud puddles, as has been the accepted generalization. Thus, control tactics aimed at immatures must consider zones larger than puddles per se. In fact, eggs are more likely to be found outside than inside puddles. Eggs can develop and larvae can emerge on mud. Larvae are then capable of three distinct modes of terrestrial displacement (two active and one passive), whereby, they can reach standing water. On mud bearing a film of water, larvae actively displace backwards by sinusoidal undulations shown to be only a slight variation of the swimming motor program. On drying mud, larvae switch to a slower and forward form of active locomotion resembling that of a crawling caterpillar. During rains, small larvae may be passively displaced by flowing rainwater so as to be deposited into puddles. These capabilities for being amphibious, along with very rapid growth and development, help explain how An. gambiae thrives in a highly uncertain and often hostile larval environment. PMID- 17139500 TI - Analysis of bioclimatic time series and their neural network-based classification to characterise drought risk patterns in South Italy. AB - A new approach to characterise geographical areas with a drought risk index (DRI) is suggested, by applying an artificial neural network (ANN) classifier to bioclimatic time series for which operational temporal units (OtUs) are defined. A climatic database, corresponding to a grid of 8 km x 8 km cells covering the Italian peninsula, was considered. Each cell is described by the time series of seven variables recorded from 1989 to 2000. Sixteen cells were selected according to land cover homogeneity and completeness of the time series data. The periodic components of the time series were calculated by means of the fast Fourier transform (FFT) method. Temporal units corresponding to the period of the sinusoidal functions most related to the data were used as OtUs. The ANN for each OtU calculates a DRI value ranging between -1 and 1. The value is interpretable as the proximity of the OtUs to one of two situations corresponding to minimum and maximum drought risk, respectively. The former set (DRI = -1) is represented by an ideal OtU with minimum values of temperatures and evapo-transpiration, and maximum values of rainfall, normalised difference vegetation index (NDVI) and soil water content. The second set (DRI = 1) is represented by the reciprocal OtU to the former one. The classification of the cells based on DRI time profiles showed that, at the scale used in this work, DRI has no dependence on land cover class, but is related to the location of the cells. The methodology was integrated with GIS (geographic information system) software, and used to show the geographic pattern of DRI in a given area at different periods. PMID- 17139501 TI - Association of CDK4 and CCND1 mRNA overexpression in laryngeal squamous cell carcinomas occurs without CDK4 amplification. AB - CDK4 is involved in the control of G1-S phase transition as a part of the CCND1/CDK4 complexes. CCNDI and CDK4 gene alterations have been implicated in the development of different tumors. CCND1 has been associated with progression in laryngeal carcinomas. CDK4 protein overexpression was described associated to CCND 1 overexpression in these tumors. However, the mechanisms implicated were not known. We analyzed CDK4 gene alterations and mRNA expression in a series of carcinomas of the larynx, and the results were compared to CCND1 expression and clinicopathological characteristics of the patients. CDK4 mRNA was overexpressed in 42 out of 60 tumors (70%) associated with CCND1 mRNA overexpression because 15 out of 16 cases with high CCND1 levels showed simultaneous increased levels of CDK4 mRNA (p = 0.023) and 12 (87%) of the tumors overexpressing both genes were in stage 4. No CDK4 gene amplifications, rearrangements, or mutations were detected in any of the tumors, including 24 overexpressed cases. These findings confirm that CDK4 overexpression is a frequent phenomenon in laryngeal carcinomas, which occurs at the transcriptional level but is related neither to gene amplification nor to gene mutation, and suggest that cooperation with CCND1 may be involved in the progression of laryngeal tumors. PMID- 17139502 TI - CT-guided needle biopsies of bone and soft tissue tumors: a pathologist's perspective. PMID- 17139503 TI - Diagnosis of superior labral lesions: comparison of noncontrast MRI with indirect MR arthrography in unexercised shoulders. AB - OBJECTIVE: To prospectively compare the accuracy of noncontrast magnetic resonance imaging (MRI) with indirect MR arthrography (I-MRa) of unexercised shoulders for diagnosis of superior glenoid labral lesions. MATERIALS AND METHODS: Institutional Review Board approval and patient informed consent were obtained for this prospective study. Superior labral findings on shoulder MRI and unexercised I-MRa studies of 104 patients were correlated with findings at arthroscopic shoulder surgery. Two musculoskeletal radiologists independently reviewed the two sets of MR images while blinded to arthroscopic results. For each radiologist, the McNemar test was used to detect statistically significant differences between techniques. RESULTS: The superior labrum was intact in 24 and abnormal in 80 subjects. For detection of superior labral lesions by each radiologist, I-MRa was more sensitive (84-91%) than MRI (66-85%), with statistically significant improvement in sensitivity for one reader (p = 0.003). However, I-MRa was less specific (58-71%) than MRI (75-83%). Overall, accuracy was slightly improved on I-MRa (78-86%) compared with MRI (70-83%), but this difference was not statistically significant for either reader. CONCLUSION: Compared with noncontrast MRI, I-MRa was more sensitive for diagnosis of superior glenoid labral lesions. However, the diagnostic value of I-MRa in shoulders remaining at rest is potentially limited by decreased specificity of the technique. PMID- 17139504 TI - Solitary intramuscular myxoma with monostotic fibrous dysplasia as a rare variant of Mazabraud's syndrome. AB - The rare coexistence of intramuscular myxoma (IM) and fibrous dysplasia (FD) is known as Mazabraud's syndrome. IM tends to occur multifocally and is associated most frequently with polyostotic FD in Mazabraud's syndrome. We present an extremely rare combination of a solitary IM and monostotic FD as a variant of Mazabraud's syndrome, and discuss the importance of recognizing this rare coexistence for appropriate management of the patient. PMID- 17139505 TI - Advanced imaging in rheumatoid arthritis. Part 1: synovitis. AB - Rheumatoid arthritis (RA) is a chronic and progressive inflammatory disorder primarily affecting the synovium. We now recognise that conventional radiographic images show changes of rheumatoid arthritis long after irreversible joint damage has occured. With the advent of powerful disease-modifying drugs, there is a need for early demonstration of rheumatoid arthritis and a need to monitor progress of the disease and response to therapy. Advanced imaging techniques such as ultrasound and MRI have focussed on the demonstration and quantification of synovitis and erosions and allow early diagnosis of RA. The technology to quantify synovitis and erosions is developing rapidly and now allows change in disease activity to be assessed. However, problems undoubtedly exist in quantification techniques, and this review serves to highlight them. Much of the literature on advanced imaging in RA appears in rheumatological journals and may not be familiar to radiologists. This review article aims to increase the awareness of radiologists about this field and to encourage them to participate and contribute to the ongoing development of these modalities. Without this collaboration, it is unlikely that these modalities will reach their full potential in the field of rheumatological imaging. This review is in two parts. The first part addresses synovitis imaging. The second part will look at advanced imaging of erosions in RA. PMID- 17139506 TI - Calculation of rotational deformity in pediatric supracondylar humerus fractures. AB - OBJECTIVE: Supracondylar humerus fractures (SCHF) are common in the pediatric population. Cubitus varus deformity (CVD) is the most common long-term complication of SCHFs and may lead to elbow instability and deficits in throwing or extension. Distal fragment malrotation in the axial plane disposes to fragment tilt and CVD; however, no simple method of assessing fracture malrotation exists. This study tested a mathematical method of measuring axial plane malrotation in SCHFs based on plain radiographs. DESIGN: A pediatric SCHF model was made, and x rays were taken at known intervals of rotation. Five independent, blinded observers measured these films. Calculated rotation for each data set was compared to the known rotation. The identical protocol was performed for an aluminum phantom. RESULTS: The reliability and agreement of the rotation values were good for both models. CONCLUSIONS: This method is a reliable, accurate, and cost-effective means of calculating SCHF distal fragment malrotation and warrants clinical application. PMID- 17139507 TI - Introduction of a disulfide bridge enhances the thermostability of a Streptomyces olivaceoviridis xylanase mutant. AB - Substitution of the N-terminus of Streptomyces olivaceoviridis xylanase XYNB to generate mutant TB has been previously shown to increase the thermostability of the enzyme. To further improve the stability of this mutant, we introduced a disulfide bridge (C109-C153) into the TB mutant, generating TS. To assess the effect of the disulfide bridge in the wild-type enzyme, the S109C-N153C mutation was also introduced into XYNB, resulting in XS. The mutants were expressed in Pichia pastoris, the recombinant enzymes were purified, and the effect of temperature and pH on enzymatic activity was characterized. Introduction of the disulfide bridge (C109-C153) into XYNB (XS variant) and TB (TS variant) increased the thermostability up to 2.8-fold and 12.4-fold, respectively, relative to XYNB, after incubation at 70 degrees C, pH 6.0, for 20 min. In addition, a synergistic effect of the disulfide bridge and the N-terminus replacement was observed, which extended the half-life of XYNB from 3 to 150 min. Moreover, XS and TS displayed better resistance to acidic conditions compared with the respective enzymes that did not contain a disulfide bridge. PMID- 17139508 TI - Quantification of host-specific Bacteroides-Prevotella 16S rRNA genetic markers for assessment of fecal pollution in freshwater. AB - Based on the comparative 16S rRNA gene sequence analysis of fecal DNAs, we identified one human-, three cow-, and two pig-specific Bacteroides-Prevotella 16S rRNA genetic markers, designed host-specific real-time polymerase chain reaction (real-time PCR) primer sets, and successfully developed real-time PCR assay to quantify the fecal contamination derived from human, cow, and pig in natural river samples. The specificity of each newly designed host-specific primer pair was evaluated on fecal DNAs extracted from these host feces. All three cow-specific and two pig-specific primer sets amplified only target fecal DNAs (in the orders of 9-11 log(10) copies per gram of wet feces), showing high host specificity. This real-time PCR assay was then applied to the river water samples with different fecal contamination sources and levels. It was confirmed that this assay could sufficiently discriminate and quantify human, cow, and pig fecal contamination. There was a moderate level of correlation between the Bacteroides-Prevotella group-specific 16S rRNA gene markers with fecal coliforms (r (2) = 0.49), whereas no significant correlation was found between the human specific Bacteroides 16S rRNA gene with total and fecal coliforms. Using a simple filtration method, the minimum detection limits of this assay were in the range of 50-800 copies/100 ml. With a combined sample processing and analysis time of less than 8 h, this real-time PCR assay is useful for monitoring or identifying spatial and temporal distributions of host-specific fecal contaminations in natural water environments. PMID- 17139509 TI - Expression of citrate permease gene of plasmid pCM1 isolated from Lactococcus lactis subsp. lactis biovar diacetylactis NIAI N-7 in Lactobacillus casei L-49-4. AB - Recombinant vector pJLECit (8,232 bp) was constructed using citrate permease gene contained in the 3,919-bp fragment of plasmid pCM1 (8,280 bp) isolated from Lactococcus lactis subsp. lactis biovar diacetylactis NIAI N-7, repA and ori from pLU1, and pMB1 ori and the erythromycin resistance gene from pJIR418. Lactobacillus casei L-49-4 (plasmid-free mutant of strain L-49) harboring the constructed pJLECit converted citrate into diacetyl/acetoin. Citrate uptake rate of resting cells was the highest at pH 5.5 and 10 mM citrate concentration. Diacetyl formation activity by the cell-free extracts of Lb. casei L-49-4 (pJLECit) grown in de Man-Rogosa-Sharpe (MRS) broth was higher than that of cells grown in MRS broth without citrate. On the other hand, diacetyl reductase activity of cells grown in MRS broth was lower than that of cells grown in MRS broth without citrate. PMID- 17139510 TI - [Therapeutic splitting in gallstones]. PMID- 17139511 TI - Dynamic control of the central pattern generator for locomotion. AB - We show that an ongoing locomotor pattern can be dynamically controlled by applying discrete pulses of electrical stimulation to the central pattern generator (CPG) for locomotion. Data are presented from a pair of experiments on biological (wetware) and electrical (hardware) models of the CPG demonstrating that stimulation causes brief deviations from the CPG's limit cycle activity. The exact characteristics of the deviation depend strongly on the phase of stimulation. Applications of this work are illustrated by examples showing how locomotion can be controlled by using a feedback loop to monitor CPG activity and applying stimuli at the appropriate times to modulate motor output. Eventually, this approach could lead to development of a neuroprosthetic device for restoring locomotion after paralysis. PMID- 17139512 TI - Bipedal locomotion: toward unified concepts in robotics and neuroscience. AB - This review is the result of a joint reflection carried out by researchers in the fields of robotics and automatic control on the one hand and neuroscience on the other, both trying to answer the same question: what are the functional bases of bipedal locomotion and how can they be controlled? The originality of this work is to synthesize the two approaches in order to take advantage of the knowledge concerning the adaptability and reactivity performances of humans and of the rich tools and formal concepts available in biped robotics. Indeed, we claim that the theoretical framework of robotics can enhance our understanding of human postural control by formally expressing the experimental concepts used in neuroscience. Conversely, biological knowledge of human posture and gait can inspire biped robot design and control. Therefore, both neuroscientists and roboticists should find useful information in this paper. PMID- 17139513 TI - [Unilateral macular hemorrhage of unknown origin]. PMID- 17139514 TI - Identification of label-retaining cells in nasopharyngeal epithelia and nasopharyngeal carcinoma tissues. AB - Adult stem cells can be identified by label-retaining cell (LRC) approach based on their ability to retain nucleoside analog, such as bromodeoxyuridine (BrdU). We hypothesized that mouse nasopharynx contains a small population of epithelial stem/progenitor cells that may be detected by the LRC technique. To identify LRCs in mice nasopharyngeal epithelia, neonatal mice were intraperitoneally injected with BrdU twice daily for 3 consecutive days. After an 8-week chase, long-term BrdU-labeled LRCs (approximately 2% of cells) were detected in the adult mice nasopharyngeal epithelia by immunostaining with BrdU antibody and some of LRCs (approximately 12% of cells) were found to be recruited into the S phase of cell cycle with an additional radioactive thymidine-labeling technique, indicating that the stem cells also divide, most likely asymmetrically. To further investigate whether the LRCs existed in human nasopharyngeal carcinoma (NPC) tissues, three NPC cell lines (5-8F, 6-10B and TMNE) were labeled with BrdU in vitro and then individually engrafted into the back of nude mice, which developed tumors. Again, label-retaining stem cells were found in all the three kinds of NPC xenograft tumors (approximately 0.3% of cells), around 16% of which were also labeled with radioactive thymidine. Thus, this study has demonstrated for the first time the presence of epithelial LRCs in mouse nasopharynx and human NPC tissues and these stem-like LRCs are not completely quiescent, as they will be recruited into the cell cycle to participate physiological or pathological process at any moment. More importantly, our data showed that NPC also contained stem cells, which are most likely the cause for NPC spread, metastasis and recurrence. PMID- 17139515 TI - Creating a cardiac pacemaker by gene therapy. AB - While electronic cardiac pacing in its various modalities represents standard of care for treatment of symptomatic bradyarrhythmias and heart failure, it has limitations ranging from absent or rudimentary autonomic modulation to severe complications. This has prompted experimental studies to design and validate a biological pacemaker that could supplement or replace electronic pacemakers. Advances in cardiac gene therapy have resulted in a number of strategies focused on beta-adrenergic receptors as well as specific ion currents that contribute to pacemaker function. This article reviews basic pacemaker physiology, as well as studies in which gene transfer approaches to develop a biological pacemaker have been designed and validated in vivo. Additional requirements and refinements necessary for successful biopacemaker function by gene transfer are discussed. PMID- 17139516 TI - Removal of eye movement artefacts from single channel recordings of retinal evoked potentials using synchronous dynamical embedding and independent component analysis. AB - A system is described for the removal of eye movement and blink artefacts from single channel pattern reversal electroretinogram recordings of very poor signal to-noise ratios. Artefacts are detected and removed by using a blind source separation technique based on the jadeR independent component analysis algorithm. The single channel data are arranged as a series of overlapping time-delayed vectors forming a dynamical embedding matrix. The structure of this matrix is constrained to the phase of the stimulation epoch: the term synchronous dynamical embedding is coined. A novel method using a marker channel with a non-independent synchronous feature is employed to identify the single most relevant source estimation for reconstruction and signal recovery. This method is non-lossy, all underlying signal being recovered. In synthetic datasets of defined noise content and in standardised real data recordings, the performance of this technique is compared to conventional fixed-threshold hard-limit rejection. The most significant relative improvements are achieved when movement and blink artefacts are greatest: no improvement is demonstrable for the random noise only situation. PMID- 17139517 TI - Study of discriminant analysis applied to motor imagery bipolar data. AB - We present a study of linear, quadratic and regularized discriminant analysis (RDA) applied to motor imagery data of three subjects. The aim of the work was to find out which classifier can separate better these two-class motor imagery data: linear, quadratic or some function in between the linear and quadratic solutions. Discriminant analysis methods were tested with two different feature extraction techniques, adaptive autoregressive parameters and logarithmic band power estimates, which are commonly used in brain-computer interface research. Differences in classification accuracy of the classifiers were found when using different amounts of data; if a small amount was available, the best classifier was linear discriminant analysis (LDA) and if enough data were available all three classifiers performed very similar. This suggests that the effort needed to find regularizing parameters for RDA can be avoided by using LDA. PMID- 17139518 TI - [Tumors of the foot skeleton]. AB - About 3-4% of all tumors and tumor-like lesions of the skeleton are located in the foot. Many of these lesions have a predilection for certain locations, so that the spectrum of entities occurring in the foot differs from the rest of the skeleton. Despite the fact that practically any entity can occur in the foot in rare cases, taken together the ten most frequent lesions make up for the vast majority of tumors and tumor-like lesions of the foot. The differential diagnosis of these lesions follows the general principles that apply in the rest of the skeleton. It is based on the analysis of the lesion's X-ray morphology and location, the patient's age, and in certain entities, the MR morphology. This article describes the most important tumors and tumor-like lesions of the foot, their differential diagnosis, and the principles of local staging. PMID- 17139520 TI - Biliary peritonitis for duct of Luschka bile leak after laparoscopic cholecystectomy performed with a 10-mm harmonic scalpel. PMID- 17139521 TI - [Schizophrenia, human genetics and genetic counselling. Human genetic counselling as part of the psychiatric/psychotherapeutic treatment concept]. AB - The human genome project has substantially increased our knowledge about the genetic basis of psychiatric diseases. In daily clinical practice the physician is asked about the diagnosis of genetically dependent diseases with an increased psychiatric risk, particularly schizophrenia group disorders, about the recurrence risk of psychiatric diseases in the relatives and children of the patients, on the use of psychopharmaceuticals during pregnancy and their potential consequences for the offspring, as well as psychopharmacogenetics. These questions will be dealt with in this contribution. At present, they receive too little attention, although genetic counselling might play an important role within the framework of psychiatric treatment, especially for schizophrenia. Such counselling should be seen as a component of the psychiatric/psychotherapeutic treatment concept and performed according to human genetic guidelines. PMID- 17139522 TI - Correlating EPR and X-ray structural analysis of arsenite-inhibited forms of aldehyde oxidoreductase. AB - Two arsenite-inhibited forms of each of the aldehyde oxidoreductases from Desulfovibrio gigas and Desulfovibrio desulfuricans have been studied by X-ray crystallography and electron paramagnetic resonance (EPR) spectroscopy. The molybdenum site of these enzymes shows a distorted square-pyramidal geometry in which two ligands, a hydroxyl/water molecule (the catalytic labile site) and a sulfido ligand, have been shown to be essential for catalysis. Arsenite addition to active as-prepared enzyme or to a reduced desulfo form yields two different species called A and B, respectively, which show different Mo(V) EPR signals. Both EPR signals show strong hyperfine and quadrupolar couplings with an arsenic nucleus, which suggests that arsenic interacts with molybdenum through an equatorial ligand. X-ray data of single crystals prepared from EPR-active samples show in both inhibited forms that the arsenic atom interacts with the molybdenum ion through an oxygen atom at the catalytic labile site and that the sulfido ligand is no longer present. EPR and X-ray data indicate that the main difference between both species is an equatorial ligand to molybdenum which was determined to be an oxo ligand in species A and a hydroxyl/water ligand in species B. The conclusion that the sulfido ligand is not essential to determine the EPR properties in both Mo-As complexes is achieved through EPR measurements on a substantial number of randomly oriented chemically reduced crystals immediately followed by X-ray studies on one of those crystals. EPR saturation studies show that the electron transfer pathway, which is essential for catalysis, is not modified upon inhibition. PMID- 17139526 TI - Paroxysmal muscle weakness: the familial periodic paralyses. AB - The familial periodic paralyses (PP) were commonly considered to be benign diseases since frequency and severity of the paralytic attacks decrease in adulthood. However, with increasing age, a third of the patients develop permanent weakness and muscle degeneration with fatty replacement. Another complication, cardiac arrhythmia, can result from the dyskalemia during paralytic attacks. The familial PP are typical dominant ion channelopathies: the function of the mutant muscular channel is compensated in the interictal state but defective under certain conditions which then cause flaccid weakness. A triggering factor is the level of serum potassium, the extracellular ion decisive for membrane excitability. In hyper- and hypokalemic periodic paralysis, the mutations are specifically located in the voltage-gated sodium and calcium channels which are essential for action potential generation or excitation contraction coupling. The common mechanism for the membrane inexcitability during paralytic attacks is a transient membrane depolarization that inactivates the sodium channels which are then no longer available for action potential generation. For the third PP type, the Andersen syndrome, the responsible gene is also expressed in cardiac muscle, and, independently of paralytic attacks, the hazard of ventricular arrhythmias is inherent. The gene product, an inwardly rectifying potassium channel, is responsible for maintaining the resting membrane potential, and all known mutations cause dominant-negative effects on the tetrameric channel complexes. In this article the clinical consequences of the mutations and the therapeutic strategies for all three types of PP are reported. PMID- 17139527 TI - Vladimir Mikhailovich Bekhterev (1857-1927). PMID- 17139528 TI - Dilute triamcinolone-assisted peeling of the internal limiting membrane in cases of diffuse diabetic macular oedema. AB - PURPOSE: In this study we evaluated the efficiency of diluted triamcinolone particles in peeling of the internal limiting membrane (ILM) in cases of diffuse diabetic macular oedema. METHODS: A prospective observational study of thirteen consecutive patients with diffuse diabetic macular oedema who had undergone ILM peeling aided by use of a diluted (1 mg mL(-1)) suspension of triamcinolone particles (0.05 mg). The results were assessed by measurement of visual acuity and intraocular pressure, by ophthalmoscopy, and by fluorescein angiography. RESULTS: ILM peeling in all eyes could be performed easily and completely with the aid of triamcinolone particles homogeneously dispersed over the macular ILM. During the peeling procedure, triamcinolone particles resulted in clear contrast between the peeled and unpeeled ILM thus enabling us to easily find the edge of the ILM and to be able to continue peeling at further attempts. In twelve of the thirteen eyes the macular oedema was completely resolved or reduced. Visual acuity improved by at least two lines in nine eyes (70%). The intraocular pressure increase did not exceed 21 mmHg in any of the patients. CONCLUSIONS: The diluted triamcinolone suspension enables good visualisation of the ILM during the ILM peeling procedure in cases of diffuse diabetic macular oedema. Use of a diluted suspension may reduce the risk of intraocular pressure increase and the potential toxicity of triamcinolone. PMID- 17139529 TI - Enhancement of the bulbocavernosus reflex during intraoperative neurophysiological monitoring through the use of double train stimulation: a pilot study. AB - OBJECTIVE: Electrophysiological bulbocavernosus reflex (BCR) testing, during surgeries in which the constituent neural components are at risk, might supplement other low sacral (S2-4) stimulation/recording techniques. However, intraoperative BCR is not always reliably implemented. We proposed to analyze BCR signals in five surgical patients monitored with the novel application of double train stimulation (DTS) to determine if the potential could be enhanced. METHODS: We prospectively planned a regime of DTS BCR with a series of intertrain delays in five monitored patients at risk for low sacral neural injury. Patients were maintained with propofol, opiate infusion, and low inhalant anesthesia without muscle relaxant. Cutaneous sensory nerves of the penis (or clitoris) were stimulated using two consecutive pulse trains (DTS). Intertrain delays were 75, 100, 125, 150, 175, 200, and 250 ms. For BCR recording, uncoated paired wires were inserted into the external anal sphincter (EAS) bilaterally. For each trial, waveform amplitude, duration, and turn count measures for the first (single train) and second (double train) response were recorded. Percent increase/decrease of the second train response compared to the first train response was calculated. RESULTS: There was at least a 30% increase in measures of amplitude, turn count, and duration of the second train response in 22/28, 22/28, and 14/28 of the total trials respectively. There was an insufficient number of independent observations to determine statistical significance. CONCLUSIONS: Intraoperative BCR is currently obtained with some difficulty using pulse train stimulation. Our preliminary evidence has identified BCR waveform enhancement using DTS and suggests that the reliability of intraoperative BCR acquisition may be further improved by the addition of this technique. Our data are insufficient to define the best intertrain interval. PMID- 17139530 TI - Development of a tightly regulated and highly inducible ecdysone receptor gene switch for plants through the use of retinoid X receptor chimeras. AB - Chemical inducible gene regulation systems provide essential tools for the precise regulation of transgene expression in plants and animals. Recent development of a two-hybrid ecdysone receptor (EcR) gene regulation system has solved some of the drawbacks that were associated with the monopartate gene switch. To further improve the versatility of the two-hybrid EcR gene switch for wide spread use in plants, chimeras between Homo sapiens retinoid X receptor (HsRXR) and insect, Locusta migratoria RXR (LmRXR) were tested in tobacco protoplasts as partners with Choristoneura fumiferana EcR (CfEcR) in inducing expression of the luciferase reporter gene. The RXR chimera 9 (CH9) along with CfEcR, in a two-hybrid format gave the best results in terms of low-background expression levels in the absence of ligand and high-induced expression levels of the reporter gene in the presence of nanomolar concentrations of the methoxyfenozide ligand. The performance of CH9 was further tested in corn and soybean protoplasts and the data obtained was compared with the other EcR switches that contained the wild-type LmRXR or HsRXR as EcR partners. In both transient expression studies and stable transformation experiments, the fold induction values obtained with the CH9 switch were several times higher than the values obtained with the other EcR switches containing LmRXR or HsRXR. The new CfEcR two-hybrid gene switch that uses the RXR CH9 as a partner in inducing reporter gene expression provides an efficient, ligand-sensitive and tightly regulated gene switch for plants. PMID- 17139531 TI - Peculiar behavior of distinct chromosomal DNA elements during and after development in the dicyemid mesozoan Dicyema japonicum. AB - The dicyemid mesozoans are obligate parasites that inhabit the cephalopod renal appendage. Dicyemids have a simple body, consisting of approximately 30 cells: one long cylindrical axial cell contains intracellular stem cells (called axoblast), from which embryos are derived, and is surrounded by some 30 peripheral somatic cells. Somatic cells divide at most eight times in their life span, and never divide after differentiation. During early somatic cell development, numerous unique DNA sequences are first amplified and then eliminated, in the form of extrachromosomal circular DNA, leading to genome reduction. In this study we demonstrate that the remaining sequences, single-copy genes and repetitive sequences, have very different fates. Single-copy genes, such as beta-tubulin, are initially amplified, presumably via endoreduplication, but subsequently decrease in copy number through development, suggesting that the whole genome is initially amplified and then the amplified DNAs are simply diluted in successive cell divisions, with little DNA replication. In contrast, repetitive sequences are maintained even in terminally differentiated somatic cell nuclei. Considering the increasing intensity of in-situ hybridization, incorporation of BrdU, and a general correlation between nuclear content and cell size, those repetitive sequences must be selectively endoreplicated in the peripheral cell nucleus, concomitant with the increase of cell size. The biological significance of this mechanism is discussed as a unique dicyemid adaptation to parasitism. PMID- 17139532 TI - Comparison of the chicken and zebra finch Z chromosomes shows evolutionary rearrangements. AB - Using fluorescent in-situ hybridization (FISH) of zebra finch (Taeniopygia guttata) bacterial artificial chromosome (BAC) clones, we determined the chromosomal localizations of 14 zebra finch genes that are Z-linked in chickens: ATP5A1, CHD1, NR2F1, DMRT1, PAM, GHR, HSD17B4, NIPBL, ACO1, HINT1, SMAD2, SPIN, NTRK2 and UBE2R2. All 14 genes also map to the zebra finch Z chromosome, indicating substantial conservation of gene content on the Z chromosome in the two avian lineages. However, the physical order of these genes on the zebra finch Z chromosome differed from that of the chicken, in a pattern that would have required several inversions since the two lineages diverged. Eight of 14 zebra finch BAC DNA showed cross-hybridization to the W chromosome, usually to the entire W chromosome, suggesting that repetitive sequences are shared by the W and Z chromosomes. These repetitive sequences likely evolved in the finch lineage after it diverged from the Galliform lineage. PMID- 17139533 TI - Origin of the roots of the accessory nerve in the goitred gazelle and Tuj sheep. PMID- 17139534 TI - Morphometric study of the intraorbital muscles (musculi bulbi) in New Zealand rabbit. AB - Eight female New Zealand rabbits were used. The bulbus oculi was removed bilaterally from orbita then intraorbital muscles were revealed by dissection and their length and breadth and the distance of the muscle insertion to the corneal limbus were measured. Junction formations of the insertion tendon of intraorbital muscle to the sclera were identified. PMID- 17139536 TI - Evaluation of a commercially available human serum amyloid A (SAA) turbidimetric immunoassay for determination of feline SAA concentration. AB - Serum amyloid A (SAA) is an acute-phase protein in cats likely to be useful for diagnosing and monitoring inflammatory diseases, especially if rapid, reliable and automated assays can be made available. A commercially available automated human SAA turbidimetric immunoassay (SAA-TIA) was evaluated for determination of SAA in cats. Intra-assay and inter-assay imprecisions were in the ranges 2.1-9.9% and 7.0-12.5%, respectively, and without significant inaccuracy. Eighty-eight cats were divided into groups according to (A) the presence or absence of an acute-phase response (APR) (n = 23 and 65, respectively) and (B) clinical diagnosis (clinically healthy cats, cats diagnosed with inflammatory/infectious diseases, endocrine/metabolic diseases, neoplastic diseases, and miscellaneous disorders (n=43, 13, 8, 4 and 20, respectively)). The observed SAA concentrations were, as expected, different for (A) cats with and without an APR and (B) cats with inflammatory/infectious diseases compared to other diagnostic groups, except neoplastic diseases. In conclusion, the SAA concentration in cats could be measured reliably using the commercially available TIA designed for measuring human SAA, which should facilitate implementation of the parameter for routine diagnostic purposes. PMID- 17139535 TI - REP-PCR analysis of Pasteurella multocida isolates from wild and domestic animals in India. AB - Repetitive extragenic palindromic sequence-based PCR (REP-PCR) was used to characterize 67 field isolates of Pasteurella multocida originating from different animal species and geographical regions of India. REP-PCR was found to be rapid and reproducible (three repeats were done). These isolates yielded different 23 profiles which were clustered into eight groups. The discrimination index was moderate (D value 0.83). Somatic and antigenic typing of the isolates did not reveal any correlation with REP-PCR profiles. There was no host-specific, type-specific, region-specific or pathenogenicity-specific pattern. The REP profiles of isolates obtained from wild animals were similar to those obtained from domestic animals. Two common bands were present in all the isolates irrespective of somatic or antigenic types. The results were not comparable with earlier findings, which had shown high discrimination index and correlation with disease presentation. PMID- 17139537 TI - Immune responses and protective efficacy of binary ethylenimine (BEI)-inactivated bluetongue virus vaccines in sheep. PMID- 17139539 TI - Occurrence of fungi from conjunctiva of healthy horses in Tuscany, Italy. PMID- 17139540 TI - Hepatic lipidosis in a captive North American porcupine (Erethizon dorsatum). PMID- 17139538 TI - Optimized transduction of canine paediatric CD34(+) cells using an MSCV-based bicistronic vector. AB - We have used a murine MSCV-based bicistronic retroviral vector, containing the common gamma chain (gammac) and enhanced green fluorescent protein (EGFP) cDNAs, to optimize retroviral transduction of canine cells, including an adherent canine thymus fibroblast cell line, Cf2Th, as well as normal canine CD34(+) bone marrow (BM) cells. Both canine cell types were shown to express Ram-1 (the amphotropic retroviral receptor) mRNA. Supernatants containing infectious viruses were produced using both stable (PA317) and transient (Phoenix cells) amphotropic virus producer cell lines. Centrifugation (spinfection) combined with the addition of polybrene produced the highest transduction efficiencies, infecting approximately 75% of Cf2Th cells. An average of 11% of highly enriched canine CD34(+) cells could be transduced in a protocol that utilized spinfection and plates coated with the fibronectin fragment CH-296 (Retronectin). Indirect assays showed the vector-encoded canine gammac cDNA produced a gammac protein that was expressed on the cell surface of transduced cells. This strategy may result in the transduction of sufficient numbers of CD34(+) BM cells to make the treatment of canine X-linked severe combined immunodeficiency and other canine genetic diseases feasible. PMID- 17139541 TI - Endocrine dysfunction in chronic severe ehrlichiosis with or without babesiosis in dogs. PMID- 17139542 TI - A comparative study on oxidative stress in dogs infected with Ehrlichia canis with or without concurrent infection with Babesia gibsoni.